PMID- 16338492 TI - A method for rapid mouse siderocyte enrichment. AB - OBJECTIVE: Iron overload is a key contributor to the pathogenesis of multiple disorders including the sideroblastic anemias. The specific iron compounds present in tissues or cells that are the target of iron deposition remain poorly understood, but there is evidence that some forms are magnetically active. We have developed a simple and specific method to purify iron-overloaded red blood cells using magnetic affinity columns. Here we describe this method and characterize purified Sod2-deficient siderocytes. MATERIALS AND METHODS: RBC derived from mice transplanted with Sod2-deficient hematopoietic stem cells served as a source of iron-laden cells. Purification was based upon the observation that iron deposits in Sod2-deficient cells are "magnetically susceptible" and allow for retention of iron-laden cells in a strong magnetic field. Peripheral blood from Sod2-deficient chimeric mice was passed through magnetic separation columns; iron-overloaded cells were eluted and characterized by flow cytometry, Western blot, and microscopy. RESULTS: We were able to purify 2.8% of the total red cells as iron-laden siderocytes. The magnetically purified Sod2-deficient cells were predominantly identified as reticulocytes. They had numerous siderotic granules, produced enhanced levels of reactive oxygen species, and showed increased protein oxidative damage, mitochondrial enrichment, and mitochondrial hyperpolarization. CONCLUSIONS: Our method can be used to purify iron-laden cells as well as iron-associated subcellular fractions prepared from iron-loaded tissues, allowing elucidation of the structure, location, and protein composition of such iron deposits. This data will help develop our understanding of the pathogenesis of SA and other disorders characterized by iron overload. PMID- 16338493 TI - Expression of CD27-CD70 on early B cell progenitors in the bone marrow: implication for diagnosis and therapy of childhood ALL. AB - OBJECTIVE: CD27, a member of the TNF receptor family, plays an important role in lymphoid proliferation, differentiation and apoptosis. This study addresses the expression of CD27 and its ligand, CD70, in children with acute lymphoblastic leukemia (ALL) and the possible role of this receptor-ligand pair in the pathogenesis of ALL. PATIENTS AND METHODS: Expression of CD27 and CD70 was evaluated with three-color flow cytometry in blood and bone marrow (BM) samples in children with ALL and controls. The biological role of these molecules on leukemic cell proliferation was studied in an in vitro culture system. RESULTS: The expression of the membrane bound CD27, as well as membrane bound CD70, on CD19(+) cells in the BM was significantly increased in ALL children compared to the expression found in the controls. Importantly, a substantial reduction in the in vitro proliferation of leukemic cells could be observed when the leukemic cells were cultured in presence of a blocking anti-human CD70 monoclonal antibody. The level of soluble CD27 (sCD27) in serum was also investigated and found to be significantly elevated in leukemic children as compared to healthy children. CONCLUSION: The high expression of CD27 and CD70 on ALL cells may represent an amplification of the normal CD27-CD70 expression present on early B cell progenitors. Our finding suggests that interference with CD27-CD70 interaction may represent novel treatment opportunities in ALL. Further studies are required to pin-point the role of this receptor-ligand pair in normal and malignant hematopoiesis. PMID- 16338494 TI - Increased induction of allogeneic-specific cord blood CD4+CD25+ regulatory T (Treg) cells: a comparative study of naive and antigenic-specific cord blood Treg cells. AB - OBJECTIVE: The genetic and immunological mechanism(s) responsible for the significant decrease in the incidence of graft-vs-host disease (GVHD) following HLA-disparate unrelated cord blood transplantation remains largely unknown. In this study, we investigated if cord blood (CB) CD4(+)CD25(+) T cells play a significant role in reducing the immune responses of allo-reactive CD4(+)CD25(-) T lymphocytes. METHODS: We compared CB CD4(+)CD25(-) and CD4(+)CD25(+) T cells, either naive or antigenic stimulated, to their counterparts in unmobilized adult peripheral blood (APB) with respect to genetic expression patterns, immunophenotype, suppressive activity, and mechanism(s) of suppression. RESULTS: Both naive CB and APB CD4(+)CD25(+) T cells expressed similarly elevated mRNA levels of CTLA-4, GITR, Foxp3, CD25, and elevated protein levels of CTLA-4 (p < 0.001) and GITR (p < 0.001). However, only naive APB but not CB CD4(+)CD25(+) T cells showed suppression of allogeneic responses. Stimulation of CD4(+)CD25(-) T cells by MUTZ-iDC (MUTZ-3-specific immature dendritic cells) elicited amplification of these genes and potent suppression (69% +/- 5% and 71% +/- 3% suppression, p < 0.001, CB and APB, respectively) on CD4(+)CD25(-) T cell proliferation induced by MUTZ-iDC but not by unrelated stimulators. Compared to that from unmobilized APB, a significantly higher percentage (2.7-fold +/- 0.5 fold; p < 0.002) of CD4(+)CD25(+)CTLA-4(+) T regulatory (Treg) cell subsets were induced from CB CD4(+)CD25(-) T cells following allogeneic stimulation. CONCLUSION: Our results suggest that CB CD4(+)CD25(+) Treg cells, which are induced at a higher rate by allogeneic stimulation when compared to unmobilized APB, can readily function as potent allogeneic immune suppressors and may in part contribute to the decrease in CB alloantigen recognition and activation of CB CD4(+)CD25(-) T cells. PMID- 16338495 TI - Interleukin-12 production by leukemia-derived dendritic cells counteracts the inhibitory effect of leukemic microenvironment on T cells. AB - OBJECTIVE: Acute myeloid leukemia (AML) cells are poorly immunogenic and inhibit T-cell function. AML-derived dendritic cells (AML-DCs) have better antigen presentation capacity than undifferentiated leukemic blasts, but may not be fully competent to stimulate T cells previously inhibited by leukemic cells. MATERIALS AND METHODS: AML-DCs were generated from AML cells and used to stimulate proliferation and cytokine production by T cells previously inhibited by AML cells. AML-DCs were also transfected with interleukin (IL)-12 gene by the nonviral method, nucleofection. RESULTS: Mature AML-DCs stimulated naive and, to a lesser extent, leukemic cell (LC)-cultured T cells more efficiently than their immature counterparts and their activity was mediated by IL-12. AML-DCs generated from CD14(-) AML samples (which represent 80% of total AML patients) were defective in IL-12 production and T-cell activation. Addition of exogenous IL-12 to LC-cultured T cells stimulated by CD14(-)-derived AML-DCs restored optimal interferon-gamma (IFN-gamma) production and Th1 skewing. IL-12 gene-nucleofected AML-DCs derived from CD14(-) cells produced significant amounts of IL-12, maintained leukemia-specific karyotype, DC-like phenotype, and function. When stimulated by IL-12-gene transduced CD14(-)-derived AML-DCs, LC-cultured T cells produced higher concentrations of IFN-gamma, thus maintaining a Th1 cytokine profile. CONCLUSION: IL-12 produced by AML-DCs plays a critical role in counteracting the inhibitory activity of LCs on T-cell function. IL-12 gene can be successfully expressed into AML-DCs defective in endogenous IL-12 production by using a novel nonviral method that does not modify their phenotypical, cytogenetic, and functional features. Genetically modified AML-DCs restore a near normal T-cell function. PMID- 16338496 TI - Antithymocyte globulin induces complement-dependent cell lysis and caspase dependent apoptosis in myeloma cells. AB - OBJECTIVE: Allogeneic stem cell transplantation is a potentially curative therapy for patients with multiple myeloma. Polyclonal antithymocyte globulins (ATG) or monoclonal anti-CD52 (Alemtuzumab) are included in conditioning regimens to enhance engraftment and reduce risk of severe graft-vs-host disease. Because both agents have been reported to induce depletion of B cells, we sought to investigate their cytotoxic activity on myeloma cells. MATERIALS AND METHODS: Complement-mediated and complement-independent activity of ATG-Fresenius and Alemtuzumab was investigated on four myeloma cell lines (RPMI-8226, U266, KMS-12 BM, and EJM) and bone marrow samples from six myeloma patients. Cytotoxicity was determined by staining with annexin V-fluorescein isothiocyanate and 7-amino actinomycin D followed by flow cytometry. RESULTS: ATG at a concentration of 500 microg mL(-1) induced up to 100% and 85% complement-dependent killing of myeloma cell lines and primary myeloma samples respectively. In the absence of complement ATG still could induce up to 50% and 80% apoptosis in myeloma cell lines and primary myeloma samples, respectively. Preincubation of myeloma cells with a general caspase inhibitor abrogated ATG-induced complement-independent cell death. Alemtuzumab-mediated myeloma cytotoxicity was only observed in KMS-12-BM cells, and in none of the patient samples. CONCLUSION: ATG induces marked cytotoxic activity both in myeloma cell lines and in primary myeloma samples. Further elucidation of antibodies and antigens involved may pave the way for antibody-based myeloma therapy. PMID- 16338497 TI - An effective and simple expansion system for megakaryocyte progenitor cells using a combination of heparin with thrombopoietin and interleukin-11. AB - Transfusion of ex vivo expanded megakaryocyte (MK) progenitor cells has been suggested to shorten the time of platelet recovery in the thrombocytopenia induced by radiotherapy or chemotherapy. Here, we report an effective and simple expansion system of MK progenitor cells from cord blood (CB) CD34(+) cells using a combination of thrombopoietin (TPO), interleukin (IL)-11, and heparin. When the CB CD34(+) cells were cultured in a liquid expansion system in the presence of TPO + recombination human (rh)IL-11 + heparin for 7 days, the number of CB CD34(+)/CD41a(+) cells was significantly increased compared to control groups (p < 0.05). When the suspension cells collected from 7-day liquid culture were replated in semisolid cultures, increased large MK colonies were observed in the culture with combination of TPO + IL-11 + heparin compared to those of control groups. In vivo, transfusion of CD34(+) cells expanded with TPO + IL-11 + heparin into irradiated nonobese diabetic/severe combined immunodeficient mice significantly accelerated platelet recovery. These data indicate that heparin as effective cofactor for TPO and IL-11 promotes expansion of MK progenitor cells from CB CD34(+) cells. This expansion system is simple and effective and could be used for the treatment of thrombocytopenia after radiotherapy or chemotherapy. PMID- 16338498 TI - Telomerized human bone marrow-derived cell clones maintain the phenotype of hematopoietic-supporting osteoblastic and myofibroblastic stromal cells after long-term culture. AB - OBJECTIVE: Gene transfer of the telomerase catalytic subunit (TERT) into primary human stromal cells prolonged their lifespan. However, primary human stromal cells are actually composed of adipocytes, myofibroblasts, osteoblasts, etc. Our objective was to investigate the phenotype and hematopoietic-support of the human telomerized stromal cell (HTS) in clonal level. MATERIALS AND METHODS: We established HTS clones (HTS-1 to HTS-9) from a parental population of HTSs by limiting dilution. Hematopoietic-supporting activity of the HTS clones was examined by coculturing with CD34(+) cells. RESULTS: HTS-1 to HTS-3 contained alkaline phosphatase (ALP)(+) cells, and HTS-4 to HTS-9 were composed of both ALP(+) and alpha-smooth muscle actin-positive cells. Although all HTS clones exhibited normal growth kinetics, one of the HTS clones exhibited a chromosomal abnormality. Moreover, the parental population of the HTS cells acquired an increased growth rate and anchorage independence after 4 years of culturing. Expression of hematopoietic growth factors, such as stem cell factor, angiopoietin-1, and hedgehog mRNA was detected in all HTS clones. The degree of hematopoietic progenitor support differed between the HTS clones, and the expansion level of CD34(+) cells was the highest in HTS-8. CONCLUSION: Human telomerized stromal cell clones exhibited the phenotype of hematopoietic supporting osteoblastic and myofibroblastic cells after long-term culture. Clinical application of HTS cells should be limited because of their potential for neoplastic transformation after hTERT gene transfer. HTS cells may be useful for analyzing the molecular mechanism of hematopoietic support of human stromal cells. PMID- 16338499 TI - Acceleration of endothelial-like cell differentiation from CD14+ monocytes in vitro. AB - OBJECTIVE: In vitro differentiation of endothelial cells has potential applications in vascular tissue engineering and cell-based therapy for many diseases. The objective of this study was to develop a new strategy that utilizes cytokines and lipopolysaccharide (LPS) to accelerate endothelial-like cell differentiation from peripheral blood CD14(+) monocytes. METHODS: Peripheral blood CD14(+) monocytes were purified with immunobeads and cultured with an angiogenic growth factor-rich growth medium (EGM-2) with or without initial treatment of LPS in combination of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 4 days (the day 4 cultures). The cells were then continuously cultured in EGM-2 medium for an additional 4 or 10 days (the day 8 or day 14 cultures). Cell markers were determined by flow cytometry analysis and immunofluorescence staining. Cytokine/chemokine profile was studied by Bio-Plex immunoassay. RESULTS: In the group of initial treatment of LPS in combination with GM-CSF, IL-4, and EGM-2, the majority of suspended CD14(+) monocytes were attached and changed their morphology to endothelial-like cells, which expressed high levels of endothelial cell markers CD31, von Willebrand factor, and vascular endothelial growth factor receptor-1 as well as two major endothelial tight junction proteins zonula occludens -1 and occludin in the day 8 cultures. Endothelial nitric oxide synthase expression was substantially increased. Endothelial-like cells were also able to uptake acetylated low-density lipoprotein and bind to Ulex europeus lectin. In addition, endothelial-like cells showed a unique cytokine/chemokine profile with substantial increases of macrophage inflammatory protein-1beta, IL-6, granulocyte colony-stimulating factor, and IL-8. CONCLUSION: Initial treatment of LPS in combination with GM-CSF, IL-4, and EGM-2 is an effective strategy for acceleration of endothelial-like cell differentiation from peripheral blood CD14(+) monocytes in vitro. PMID- 16338500 TI - T-cell P/E-selectin ligand alpha(1,3)fucosylation is not required for graft-vs host disease induction. AB - OBJECTIVE: Recognition of E- and P-selectins on vascular endothelium by their leukocyte glycoprotein counterreceptor P-selectin glycoprotein ligand-1 (PSGL-1) initiates and sustains leukocyte rolling, culminating in extravasation of lymphocytes from blood into organs. PSGL-1 is rendered functional by terminal glycosylation steps, which occur mainly in activated Th1 but not Th2 cells. alpha(1,3)Fucosyltransferases IV and VII control this glycosylation pathway. Mice lacking these transferases (Fuc-TIV(-/-)/Fuc-TVII(-/-)) lack functional E- and P selectin ligands. We hypothesized that Fuc-TIV(-/-)/Fuc-TVII(-/-) donor T cells might have reduced capacity to roll on vessels of inflamed target tissues and mediate graft-vs-host disease (GVHD). MATERIALS AND METHODS: We compared the ability of Fuc-TIV(-/-)/Fuc-TVII(-/-) and wild-type (WT) C57BL/6 (B6) spleen cells (SPCs) to produce GVHD in lethally irradiated major histocompatibility complex (MHC) haplotype-mismatched B6D2F1 recipients. Clinical GVHD, GVHD pathology in target organs, memory phenotype conversion, proliferation of donor T cells, and tissue and serum cytokine expression were examined. RESULTS: Surprisingly, clinical GVHD was not reduced in lethally irradiated mice receiving full haplotype MHC mismatched or matched Fuc-TIV(-/-)/Fuc-TVII(-/-) SPCs compared to those receiving WT SPCs. GVHD pathology in target organs, memory phenotype conversion, and proliferation of donor T cells were similar in both groups. However, reduced interferon-gamma was detected in liver and lung, and serum levels of tumor necrosis factor-alpha were higher in mice receiving Fuc-TIV(-/ )/Fuc-TVII(-/-) SPCs compared with WT SPCs. CONCLUSIONS: These results suggest that donor T cells, including Th1, are capable of trafficking to GVHD target tissues independently of P- and E- selectin ligand in conditioned hosts. PMID- 16338501 TI - Door-to-ECG time in patients with chest pain presenting to the ED. AB - OBJECTIVE: To describe time to electrocardiogram (ECG) acquisition, identify factors associated with timely acquisition, and evaluate the influence of time to ECG on adverse clinical outcomes. METHODS: We measured the door-to-ECG time for emergency department patients enrolled in prospective chest pain registry. Clinical outcomes were defined as occurrence of myocardial infarction or death within 30 days of the visit. RESULTS: Among patients with acute coronary syndrome (ACS), 34% and 40.9% of patients with non-ST-elevation ACS and ST-elevation myocardial infarction (STEMI), respectively, had an ECG performed within 10 minutes of arrival. A delay in ECG acquisition was only associated with an increase risk of clinical outcomes in patients with STEMI at 30 days (odds ratio, 3.95; 95% confidence interval, 1.06-14.72; P = .04). CONCLUSION: Approximately one third of patients with ACS received an ECG within 10 minutes. A prolonged door-to-ECG time was associated with an increased risk of clinical outcomes only in patients with STEMI. PMID- 16338502 TI - The effectiveness of an ED-based violence prevention program. AB - BACKGROUND: Youth violence continues to be a problem in the United States, most prominent in the inner-city minority youth population. The recurrence rate for repeat violence has been reported from 6% to 44% with a 5-year mortality of 20%. This study describes the results of a program to reduce violence recurrence based in the ED. METHODS: Patients aged 10 to 24 years who were victims of interpersonal violence (excluding child abuse, sexual assault, and intimate partner violence) were randomly enrolled in the study in level 1 trauma center. The control group was given a written list of services, and the treatment group received an assessment and case management for 6 months. Both groups were evaluated 6 and 12 months after enrollment in the study. The primary indicators of the success of the intervention were reduction of self-reported revictimization or arrest and state-reported incarceration and reinjury. The study was approved by the institutional review board. The results compared the change in treatment and control groups over the time using a combination of chi(2) and analysis of covariance. RESULTS: One hundred eighty-eight victims of interpersonal violence met the criteria and had the initial evaluation completed. By chi(2) analysis, the treatment group (96 subjects) and the control group (92 subjects) were similar in age, sex, and racial composition. The average age was 18.6 years (range, 11-24), and 82.5% were boys. Most youth were African Americans (65.4%), followed by Hispanic (31.4%), whites (1.6%), or others (1.5%). A reduction in the self-reported reinjury rate was significantly reduced over time in the treatment group (chi(2) 3.87, P = .05). There were no differences between the groups in the number of self-reported arrests, state-reported reinjuries via the trauma registry, or state-reported incarcerations (P < .05). CONCLUSIONS: The results of this study demonstrated a reduction in self-reported reinjury rate in the intervention group. Further research is needed to confirm if ED-based violence prevention programs are effective in reducing other determinants for revictimization. PMID- 16338503 TI - Can stock rotation effectively mitigate EMS medication exposure to excessive heat and cold? AB - The United States Pharmacopeia recently published a general chapter specifically addressing on-ambulance storage of medications, including a suggestion for stock rotation. This study describes the effectiveness of a simple stock rotation strategy in mitigating EMS medication exposure to excessive heat and cold. Previously collected on-ambulance temperature data from 5 US cities were randomly resampled to generate model exposures of 2 days to 6 months duration. The temperature measurements for every other 24-hour period were then set at 20 degrees C to model the rotation of medications into a controlled environment. For each model, we then determined consistency with the official United States Pharmacopeia definition of controlled room temperature. Without stock rotation, excessive heat occurred in 39.9% of the model exposures. With stock rotation, exposures to excessive heat occurred in less than 1% of northern city models and in 2.9% of the central US models. Stock rotation did not reduce heat exposures in the models for southern cities. PMID- 16338504 TI - Diagnosis of bowel and mesenteric injuries in blunt abdominal trauma: a prospective study. AB - PURPOSE: Currently, nonoperative management is the procedure of choice for solid organ injury in patients with a blunt abdominal trauma. Missed blunt bowel and mesenteric injuries (BBMIs) are possible because diagnosis is difficult. The aim of our study was to test a new algorithm for BBMI diagnosis using abdominal ultrasonography (AUS), computed tomography (CT), and diagnostic peritoneal lavage (DPL). METHODS: We reviewed cases of blunt abdominal injuries over a 10-year period, then we designed an algorithm that was prospectively tested in hemodynamically stable patients over a 2-year period. An abnormal AUS led to helical CT. When the CT showed more than 2 findings suggestive of BBMI, laparotomy was performed. In case of 1 or 2 abnormal CT findings, we performed a DPL and calculated the ratio of white blood cells (WBCs) to red blood cells (RBCs) (WBC/RBC ratio) in the lavage fluid and divided this by the WBC/RBC ratio in peripheral blood. A ratio of 1 or higher was considered positive for BBMI, and a laparotomy was immediately performed. Patients with a ratio of less than 1 were managed nonoperatively. RESULTS: In the retrospective study, 26 (1%) of 2126 patients admitted to our trauma center for blunt trauma had a BBMI, including 15 (58%) diagnosed after a median delay of 24 hours. In the prospective study, 531 patients were admitted for blunt trauma with multiple injuries, including 131 with abdominal trauma. Computed tomography was performed in 40 patients. There were 2 criteria or more of BBMI in 1 patient, 0 criteria in 27 patients (with an uneventful follow-up), and 1 or 2 criteria in 12 patients who had DPL with a median ratio of 0.82 (ranges, 0.03-9). Five patients had a ratio of 1 or higher. They underwent immediate laparotomy. In all 5 cases, BBMI was found. The 7 patients who had a ratio of less than 1 were observed in ICU and treated for extra-abdominal injuries. No BBMI injury was missed in these patients. The accuracy of the algorithm was 100% (95% confidence interval, 0.99-1.00). CONCLUSION: The proposed algorithm (based on AUS, CT, and DPL) had a high accuracy to diagnose BBMI while requiring the performance of DPL in only a few (2%) patients. PMID- 16338505 TI - The effect of treatment on the presence of abnormal heart sounds in emergency department patients with heart failure. AB - OBJECTIVE: We sought to assess the proportion of ED patients with an electronically detected S(3) or S(4), determine the relation of these heart sounds to heart failure (HF), and analyze how the proportion changes with ED treatment. METHODS: Heart sounds were assessed in ED patients with suspected HF. The presence or absence of HF and whether treatment with diuretics or vasodilators had occurred were recorded. RESULTS: Three hundred seventy-six patients had complete data. The proportion of patients with an S(3) and an S(4) was significantly higher for those with HF compared with those without (P < .001). Of 59 patients with HF evaluated before treatment, 57.6% had an S(3) and 35.6% had an S(4). For the 35 patients with HF evaluated after treatment, the proportions of both S(3) and S(4) were lower (28.6% and 8.6%, respectively; P < or = .0064). CONCLUSIONS: This study suggests the proportion of patients with an electronically detected S(3) in HF is more than 50%, and that its presence is affected by prior treatment with diuretics or vasodilators. PMID- 16338506 TI - Changes in access to primary care for Medicaid beneficiaries and the uninsured: the emergency department perspective. AB - Reductions in scope of benefits and stricter premium and co-payment policies in the Oregon Health Plan (OHP) led to a large drop in OHP enrollment. Outpatient psychiatric benefits were eliminated for approximately 25% of enrollees. One measure of access to care is ED use. We used administrative data from our ED from August 1, 2001, through June 30, 2004, comparing ED use before vs after the March 1, 2003, cutbacks. Before the cutbacks, 38% of ED visits were by OHP beneficiaries, falling to 32% afterward. Visits by the uninsured rose from 18% before to 22% afterward. The proportion of visits for psychiatric conditions covered by OHP fell from 41% [corrected] to 31% [corrected], although the proportion by uninsured patients rose from 16% to 23%. These findings suggest a worrisome reduction in access to medical care for uninsured Oregonians and unstable access for OHP enrollees, especially for behavioral health conditions. PMID- 16338507 TI - Albuterol nebulized in heliox in the initial ED treatment of pediatric asthma: a blinded, randomized controlled trial. AB - OBJECTIVE: A prospective blinded, randomized controlled trial was undertaken to compare the initial response of albuterol nebulized in heliox or control in the treatment of moderately severe asthma in children presenting to a pediatric ED. METHODS: Patients were randomized to receive heliox (n = 20) or control (n = 21). The primary outcome was to compare a modified dyspnea index score at 10 and 20 minutes after randomization. Secondary outcomes were to determine if heliox decreased admission rates or endotracheal intubation. RESULTS: There was no statistically significant difference found at 10 or 20 minutes after randomization with heliox (P = .169 and P = .062, respectively). No statistical difference in admission rate was found, and no patients required endotracheal intubation in either group. CONCLUSIONS: Our results demonstrate that albuterol nebulized with heliox offered no clinical benefit over standard therapy in the initial treatment of moderately severe asthma in the ED. PMID- 16338508 TI - Impact of helical computed tomography on the rate of negative appendicitis. AB - OBJECTIVE: To assess the impact of helical computed tomography (HCT) on the rate of negative appendicitis (NA). METHODS: A before-and-after comparison study comparing data from a prospective consecutive case series to data from a retrospective chart review. The prospective series included all patients presenting to the ED during a 19-month period, during which patients with suspected appendicitis were managed in accordance with a guideline that stipulated HCT in selected cases. The retrospective group included patients taken to the operating room (OR) with a preoperative diagnosis of appendicitis over a 4 year time frame before the use of HCT. The primary outcome variable was the rate of NA. RESULTS: During the HCT era, 104 of 310 patients, 71 (68%; 95% confidence interval [CI], 59-76) men and 33 (32%; 95% CI, 24-41) women, were taken to the OR with a diagnosis of appendicitis. Fourteen (13.5%; 95% CI, 8-21) were NA. During the pre-HCT period, 445 patients, 280 (62.9%; 95% CI, 58-67) men and 165 (37.1%; 95% CI, 33-42) women were taken to the OR with the preoperative diagnosis of appendicitis, and 66 (14.8%; 95% CI, 12-19) were NA. CONCLUSION: At the study institution, the selective use of HCT did not result in a significant decline in the rate of NA. PMID- 16338509 TI - Correlation of initial emergency department pulse oximetry values in asthma severity classes (steps) with the risk of hospitalization. AB - BACKGROUND: The oxygen saturation (OSAT) of wheezing children presenting to an emergency department has been shown to be a predictor for hospitalization. The purpose of this study is to determine if hospitalization predictive power can be increased by further stratifying this by the step severity categories (based on chronic symptoms). METHODS: Data were collected prospectively at 6 centers over a 22-month period on 1219 pediatric patients. Asthma step severity categorization was determined by chronic symptom history. Presenting ED OSAT values, extensive clinical histories (obtained in the ED and during several telephone follow-up calls by study personnel), treatments, and disposition were recorded for each study subject. RESULTS: The overall hospitalization rate was 15%. Hospitalization rates in severity step categories 1, 2, 3, and 4 were 13%, 16%, 13%, and 22% (P = .008), respectively. Hospitalization rates by presenting OSATs were 98% or higher (6%), 95% to 97% (12%), 93% to 94% (28%), 90% to 92% (45%), 85% to 89% (65%), and 80% to 84% (100%). From 95% to 100% OSAT values, hospitalization rates are similar between the severity groups. In the 93% to 94% OSAT group, the hospitalization rate is 43% in step category 4 patients, compared with 27%, 24%, and 13% for step categories 1, 2, and 3, respectively, but this difference was not statistically significant. At presenting OSAT values of 90% and below, the hospitalization rates are higher but did not differ significantly between the severity step groups. No recognizable trend was present to suggest that the hospitalization predictive value is increased by adding the step severity categories. CONCLUSIONS: The presenting OSAT is the dominant initial predictor of hospitalization. The step severity categories do not appear to provide substantial additional predictive value for hospitalization. PMID- 16338510 TI - Serial Bispectral index scores in patients undergoing observation for sedative overdose in the emergency department. AB - STUDY OBJECTIVE: Many patients who overdose on sedatives experience a declining mental status and eventually require endotracheal intubation. The goal of this study was to determine if serial bedside Bispectral index (BIS) scores monitoring can be used to detect the eventual need for intubation in overdosed patients who are undergoing observation in the ED. METHODS: This was a prospective, observational study of a convenience sample of patients who presented to the Hennepin County Medical Center ED between June and November 2002. Patients being treated and observed for a suspected sedative ingestion were eligible. Upon presentation, a Bispectral electroencephalographic probe was applied to the patient's forehead, and a BIS score was recorded at 0 and 20 minutes. The Altered Mental Status scale was used to describe the patient's clinical status. Data were collected by trained research assistants. Data are described with descriptive statistics. The mean changes in BIS score between patients who did and did not require intubation are compared with t tests, and the outcome of patients with stable vs declining BIS scores were compared with chi(2) tests. RESULTS: Seventy six patients were enrolled. The mean initial BIS score was 83.9 (95% CI, 79.7 88.1; range, 9-99). The mean change in BIS scores during the 20-minute observation period for the patients who required intubation was -13.5 (95% CI, 30.2 to 3.2) and was +6.7 (95% CI, 3.3-10.1) for those who were not intubated. Sixteen patients had an initial BIS score below 70. Of these patients, 6 were intubated. All intubations occurred during the 20 minutes, and this group had a mean initial BIS of 47.2 (95% CI, 35.6-58.8). The 10 patients with an initial BIS below 70 who were not intubated had a mean increase in BIS score of 23.3 (95% CI, 11.7-33.9) during the 20 minutes. Of the 60 patients whose first BIS score was above 70, 5 were eventually intubated during their ED treatment. The mean change in BIS was -36.4 (95% CI, -18.7 to -54.1) for the intubated patients vs +7.9 (95% CI, 4.4-11.3) for nonintubated patients during the first 20 minutes. CONCLUSION: The overdosed patients who required intubation during their ED treatment experienced a mean decrease in BIS during the first 20 minutes, compared with those who did not. Bispectral index scores monitoring may prove useful for earlier ED treatment and decision making regarding sedative overdose patients. PMID- 16338511 TI - A pilot study of cytokine levels and white blood cell counts in the diagnosis of necrotizing fasciitis. AB - STUDY OBJECTIVES: To characterize the early cytokine response of patients presenting to the emergency department with necrotizing fasciitis (NF) and to determine whether serum cytokine levels and white blood cell (WBC) counts may be useful in distinguishing NF from other severe soft-tissue infections. METHODS: White blood cell counts and cytokine levels (IL-1beta, IL-1Ra, IL-6, IL-8, IL-18, and IFN-gamma) were measured in patients presenting to the emergency department with severe soft-tissue infections and high suspicion of NF. Necrotizing fasciitis was confirmed intraoperatively and by surgical pathology. Cytokines were measured via the liquid-phase electrochemiluminescence method. RESULTS: Thirty-five patients were enrolled, 18 were diagnosed with NF, and 17 were diagnosed with cellulitis and/or abscess (CAB). On admission, patients with NF had significantly higher WBC counts and lower levels of interleukin 1beta (IL 1beta) compared with patients with CAB. There were no statistically significant differences in the levels of the other cytokines between the 2 groups. CONCLUSION: Patients with NF have higher WBC counts and lower IL-1beta levels compared with patients with CAB. PMID- 16338512 TI - ALARMED: adverse events in low-risk patients with chest pain receiving continuous electrocardiographic monitoring in the emergency department. A pilot study. AB - OBJECTIVES: Current guidelines suggest that most patients who present to an emergency department (ED) with chest pain should be placed on a continuous electrocardiographic monitoring (CEM) device. We evaluated the utility of CEM in ED patients with chest pain. METHODS: We enrolled stable patients who presented to a single ED with chest pain suspected to be ischemic in origin and who were placed on CEM. Patients were classified according to risk of poor outcome using 3 published stratification tools. Trained observers prospectively recorded number of monitored hours, alarms, changes in management, and monitor-detected adverse events (AEs). The primary outcome measure was the rate of AEs detected by CEM. Secondary outcome measures were the rate of alarms that resulted in a change in management and number of false alarms. RESULTS: We enrolled 72 patients, 56% of whom were categorized as very low-risk by Goldman risk criteria. During 371 monitored hours, we recorded 1762 alarms or 4.7 alarms per monitored hour. There were 11 AEs (0.68%; 95% CI, 0.35%-1.2%), 3 of which resulted in a change in management (0.2%; 95% CI, 0.04%-0.5%). Seven AEs were bradydysrhythmias with a heart rate of 45 or higher; the eighth patient had no change in symptoms and was given atropine for a heart rate of 32. The other 3 AEs were an untreated supraventricular tachycardia, a brief sinus pause that triggered a rate change in intravenous nitroglycerin by the patient's nurse, and a run of premature ventricular contractions after which heparin was administered. None of the 3 patients with a change in management was categorized as the lowest-risk. CONCLUSIONS: Routine CEM in low-risk ED patients with chest pain results in an excessive number of alarms, most of which require no change in management. In these patients, the benefit of CEM may be limited, and given that 99.4% of alarms were false, current CEM technology needs to be improved. PMID- 16338513 TI - Clinical guidelines, computed tomography scan, and negative appendectomies: a case series. AB - INTRODUCTION: Prior studies suggest that preoperative abdominal computed tomography (CT) scan can contribute to a low negative appendectomy rate. Our study objective was to evaluate cases of negative appendectomies for clinical criteria and CT use. METHODS: Retrospective 1-year study of all negative appendectomies at a community hospital. Data included clinical evidence for appendicitis as well as CT results if performed. RESULTS: Of 122 cases reviewed, 8 (6.6%) were negative appendectomies. Six were younger than 20 years. Four had a preoperative CT scan. Overall, 106 (87%) of 122 cases received preoperative CT and had a negative appendectomy rate of 3.8%. Sixteen cases did not have preoperative CT and had a negative appendectomy rate of 25% (odds ratio, 8.5; 95% confidence interval, 1.9-38.5; Fisher exact test P = .01). CONCLUSION: Patients who had a CT scan for suspected appendicitis had a lower rate of negative appendectomies than those who had no CT. Most of the negative appendectomies occurred in patients younger than 20 years. PMID- 16338514 TI - Prevention of high-pressure injection injuries to the hand. AB - OBJECTIVES: To identify populations at high risk for, and the usual mechanisms of injury in, high-pressure injection injuries to the hand. METHODS: A case note review of a historical cohort of 76 patients, presenting with high-pressure injections injuries to the hand over a 12-year period, collected information including sex, age, hand dominance, and occupation of the patient and mechanism of injury, when documented. RESULTS: Eighty-two percent of these injuries were work-related, affecting mainly manual workers (84%), including 13 painters, 10 mechanics, 8 farmers, and 3 water blasters. The mechanism of injury, recorded in 63%, was most commonly a ruptured hose or inadvertent gun discharge during cleaning or use. CONCLUSIONS: Preventative measures could include a targeted safety program for equipment users, engineering improvements in gun and hose design, economic incentives, and workplace legislation. PMID- 16338515 TI - The athlete's electrocardiogram. AB - The electrocardiogram performed in the competitive athlete may manifest abnormal electrocardiographic findings; these findings may indicate either normal variant syndromes as well as true cardiac pathology. The normal variant syndromes include ST-segment and T-wave abnormalities, rhythm disturbances, and intraventricular conduction delay--it must be stressed that these electrocardiographic findings are, in fact, normal variants, not indicative of underlying pathology. Other presentations in these same competitive athletes describe significant cardiac pathology, including syndromes predisposing the patient to sudden cardiac death and other potentially dangerous dysrhythmias and diagnostic of acute coronary syndrome. This article reviews the various findings in this group of patients. PMID- 16338516 TI - Body surface mapping in the ED evaluation of the patient with chest pain: use of the 80-lead electrocardiogram system. AB - Body surface mapping (BSM) is an electrocardiographic (ECG) technique that uses numerous leads on a patient's anterior and posterior chest, enabling more complete visualization of cardiac electrical activity. The rationale behind BSM is an extension of additional lead ECG. Output from BSM is displayed in a 12-lead ECG format, an 80-lead ECG format, and on color contour maps. The color contour maps can be displayed on a torso image or as a flat map. PMID- 16338517 TI - The sensitivity and specificity of the digital rectal examination for detecting spinal cord injury in adult patients with blunt trauma. AB - STUDY OBJECTIVES: The digital rectal examination (DRE) may assist physicians in detecting spinal cord injury in patients with blunt trauma. However, the test characteristics of the DRE for detecting spinal cord injury are unknown. METHODS: We conducted a retrospective review of consecutive adult patients with blunt trauma over a 2-year period. The DRE result was compared with the presence or absence of spinal cord injury at discharge. RESULTS: A total of 1032 adult patients with blunt trauma had a DRE. Of these, 54 (5.2%) had diagnoses consistent with spinal cord injury. Ninety-nine patients had decreased rectal tone, 27 of whom also had spinal cord injuries. The sensitivity, specificity, positive predictive value, and negative predictive values were 50%, 93%, 27%, and 97%, respectively. CONCLUSION: The DRE is insensitive to spinal cord injury and has a poor positive predictive value. The high specificity must be balanced against the large number of false-positive results. PMID- 16338518 TI - Thrombocytosis accompanying urinary tract infection suggests obstruction or abscess. AB - OBJECTIVE: Reactive thrombocytosis can be found in patients with different types of infections, including upper urinary tract infection (UTI). In this study, we determined whether thrombocytosis in patients with upper UTI is a random phenomenon or is related to complications associated with the UTI. METHODS: We reviewed the charts of patients admitted to the urology department with a diagnosis of upper UTI and thrombocytosis (platelet count >500 x 10(9)/L) in the years 1991 to 2003 (study group). Patients admitted to the urology department in the year 2003 with a diagnosis of upper UTI without thrombocytosis served as the control group. RESULTS: The study group consisted of 83 patients admitted to the urology department with a diagnosis of upper UTI and thrombocytosis (mean platelet count, 593 x 10(9)/L; SD, 88; range, 501-949). The control group consisted of 99 patients. An obstructed kidney was found in 54 patients (65%) of the study group and in 18 patients (18%) of the control group (P < .00001). Perinephric abscess was found in 7 patients (8%) in the study group compared with none in the control group (P < .003). The positive predictive value of thrombocytosis for kidney obstruction or abscess in patients with upper UTI was 71%. Nephrectomy of a nonfunctioning infected kidney was required in 6 patients of the study group (7%) and 2 of the control group (P = .14). In 26 patients (31%) of the study group, the finding of thrombocytosis preceded the diagnosis of the complication (by a median period of 3 days). In these patients, thrombocytosis was essentially an early marker for the complication. CONCLUSIONS: Thrombocytosis in a patient with upper UTI is not a random phenomenon. It is a marker of kidney obstruction or perinephric abscess. The finding of thrombocytosis in a patient with upper UTI should alert the attending physician to actively search for these complications. Cross-sectional imaging study (abdominal ultrasonography or computed tomography) should be performed. PMID- 16338519 TI - Pseudo-Wellens syndrome in a cocaine user. PMID- 16338520 TI - Subarachnoid hemorrhage in survivors of out-of-hospital cardiac arrest: true or not? PMID- 16338521 TI - Chronic severe dystonia after single exposure to antiemetics. PMID- 16338522 TI - Oedipism: bilateral self-enucleation. PMID- 16338523 TI - Mycotic aneurysm of superior mesenteric artery branch presenting as pulsatile abdominal mass. PMID- 16338524 TI - The effects of sorbitol on gastric emptying half-times and small intestinal transit after drug overdose. PMID- 16338525 TI - Methamphetamine-induced myocardial infarction with elevated troponin I. PMID- 16338526 TI - Nonsurgical management of symptomatic posttraumatic thoracic epidural hematoma. PMID- 16338527 TI - Pulse oximetry in the detection of hypercapnia. PMID- 16338528 TI - The growing abuse of commonly prescribed psychiatric medications. PMID- 16338529 TI - Cataract and refractive surgery: reflecting on journal impact factors and influence. PMID- 16338530 TI - Restoring accommodation: what is real and what is pseudo? PMID- 16338532 TI - Trabeculectomy versus viscocanalostomy for primary open-angle glaucoma. PMID- 16338534 TI - Investigation of postoperative endophthalmitis. PMID- 16338535 TI - New formula for calculating intraocular lens power after LASIK. PMID- 16338536 TI - Underestimation of corneal thickness by Orbscan after myopic correction. PMID- 16338537 TI - Intracameral injection of lidocaine and carbachol. PMID- 16338538 TI - Visual sensation during phacoemulsification using topical versus regional anesthesia. PMID- 16338539 TI - Retrocapsular lens fragments after uneventful phacoemulsification cataract surgery. PMID- 16338540 TI - Inhibition of lens epithelial cell migration by contact pressure. PMID- 16338541 TI - Consultation section. Cataract surgical problem. PMID- 16338552 TI - Use of trypan blue for penetrating keratoplasty. AB - Use of trypan blue for penetrating keratoplasty was developed to facilitate the procedure. Trypan blue is injected before and after the addition of 0.25 mL of an ophthalmic viscosurgical device (OVD), sodium hyaluronate, to stain the internal and external cut edge of the cornea as well as the OVD, enabling the surgeon to improve visualization of the incision and suture depth, improve alignment of host and donor tissues, and ensure that all OVD is removed. PMID- 16338553 TI - Treating astigmatism after a free laser in situ keratomileusis cap by rotating the cap. AB - Induced astigmatism and loss of best corrected visual acuity are complications of inadvertently rotating a free cap of laser in situ keratomileusis. It is possible to determine the angle of rotation from the induced astigmatism and correct the astigmatism by lifting and rotating the cap to an astigmatically neutral position. PMID- 16338554 TI - Cataract surgery in eyes with iridoschisis using the Perfect Pupil iris extension system. AB - PURPOSE: To evaluate the use of the Perfect Pupil system (Milvella Pty. Ltd.) in 3 eyes in cataract surgery with iridoschisis. SETTING: Department of Ophthalmology, University of Heidelberg, Heidelberg, Germany. METHODS: Patient 1 was an 83-year-old man with a nuclear cataract, pseudoexfoliation syndrome, and iridoschisis. He was also having systemic anticoagulation therapy. Preoperative best corrected visual acuity (BCVA) was 0.05 in the right eye and 0.08 in the left eye. Patient 2 was a 73-year-old mentally retarded man with a brunescent cataract in the right eye. Preoperative BCVA was 0.1. RESULTS: In the first patient, cataract surgery was performed under topical anesthesia in both eyes through a clear corneal incision. Intraoperative pupil size was 3.0 mm. The Perfect Pupil device was inserted manually and fixated at the iris rim, leading to pupil dilation of 6.0 mm. Uneventful cataract surgery was performed without damage to the iris. Hydrophobic acrylic foldable intraocular lenses (IOLs) were implanted with forceps. Postoperative BCVA was 0.5 in the right eye and 0.4 in the left eye. The second patient was operated on under general anesthesia. Pupil dilation was successfully achieved. The brunescent cataract was removed and a hydrophobic acrylic IOL was implanted with an injector, resulting in postoperative uncorrected visual acuity of 0.3. CONCLUSION: In iridoschisis patients, there is risk of aspiration of iris fibers during cataract surgery. In these 2 patients, this was prevented using the Perfect Pupil system. PMID- 16338555 TI - Effect of hinge location on corneal sensation and dry eye after laser in situ keratomileusis for myopia. AB - PURPOSE: To evaluate the effects of a superior or nasal hinge location on corneal sensation and dry eye after laser in situ keratomileusis (LASIK). SETTING: Magill Research Center for Vision Correction, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: This prospective randomized masked study included 47 patients having bilateral myopic LASIK surgery. The first eye was randomly assigned to have a nasal or superior hinge flap; the fellow eye had the alternate location. Visual acuity, contrast sensitivity, corneal sensation, basic secretion test, tear film breakup time, conjunctival and corneal staining, and a subjective questionnaire were evaluated preoperatively and postoperatively at 1 week and 1, 3, and 6 months. The Wilcoxon signed rank test and paired t test were used for comparison. RESULTS: Dry eye occurred with the same frequency in both groups. However, eyes with the nasal hinge had significantly better nasal sensation than those with the superior hinge (P<.05) at 1 month. CONCLUSION: Better nasal corneal sensation was found at 1 month in the nasal hinge group but there was no difference in any other parameters measured. PMID- 16338556 TI - Influence of optic edge design, optic material, and haptic design on capsular bend configuration. AB - PURPOSE: To assess and classify capsular bend configuration at the optic rim and its association with posterior capsule (PCO) and anterior capsule opacification (ACO) and capsulorhexis contraction with various intraocular lenses (IOLs). SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: A total of 659 post-cataract surgery eyes of 370 patients from different prospective randomized studies were evaluated. All eyes had standardized phacoemulsification surgery with implantation of an IOL in the capsular bag. The IOLs had a sharp or round optic edge design and were made of silicone, acrylic, or poly(methyl methacrylate) optic materials. One year postoperatively, the configuration of the capsular bend at the optic rim was assessed at the slitlamp, and standardized slitlamp images of ACO and fibrotic PCO were taken. One week, 1 year, and 3 years postoperatively, digital retroillumination images were taken for objective quantification of regeneratory PCO and the capsulorhexis area. The outcome measures were the type of capsular bend configuration, ACO score (0% to 100%), regeneratory PCO score (0 to 10), fibrotic PCO score (0 to 3), and amount of capsulorhexis contraction (mm(2)). RESULTS: Four main types of capsular bend configuration were classified: parallel, "Y," right angle, and wrapping. The right-angle type was observed in the most cases (52%). Eyes with a wrapping capsular bend configuration had significantly less PCO, more capsulorhexis contraction, and more ACO than eyes with the other configurations. Wrapping capsule configuration was seen most often (55%) in round-edged silicone IOLs that had a thin optic rim. CONCLUSION: The design and material of IOLs influenced the long-term capsular bend configuration at the optic rim. The right-angle type was the most common capsular bend configuration. Intraocular lenses with silicone optic material and a thin optic rim caused a wrapping capsule configuration and resulted in more capsulorhexis contraction and ACO, but less PCO. PMID- 16338557 TI - Laser in situ keratomileusis in patients with diabetes. AB - PURPOSE: To study the outcomes of laser in situ keratomileusis (LASIK) in patients with well-controlled diabetes mellitus. SETTING: Gimbel Eye Centres, Calgary and Edmonton, Canada. METHODS: The charts of all patients with diabetes who had LASIK surgery at the Gimbel Eye Centres were reviewed retrospectively. Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), final refraction, and complications were assessed. RESULTS: Twenty-four patients (16 men and 8 women) were identified. Mean patient age was 42 years (range 24 to 57 years). Seventeen patients had diabetes type II, and 7 had diabetes type I. Mean preoperative spherical equivalent (SE) was -4.88 +/- 2.13 diopters (D) (range +1.625 to -9.00 D). Median follow-up was 6 months (range 4 to 44 months). No eye lost BSCVA. Twenty-nine eyes (63%) achieved UCVA 20/25 or better, and 31 eyes (67%) were within +/-0.5 D of the intended refraction after the first LASIK surgery. Retreatment was required in 13 eyes (28.3%) because initial surgery was not adequate to correct the refractive error. At the last follow-up visit, 40 eyes (87%) achieved UCVA of 20/25 or better and 43 eyes (93.5%) were within +/ 0.5 D of the intended refraction. Three eyes (6.5%) developed an epithelial defect after surgery, and secondary epithelial ingrowth developed in 2 of these eyes. No advancement of diabetic retinopathy was noticed in any eye at the end of the follow-up period. CONCLUSIONS: Laser in situ keratomileusis surgery was safely performed in patients with well-controlled diabetes. Enhancement may often be required for optimal correction. PMID- 16338558 TI - Photorefractive keratectomy and laser in situ keratomileusis in refractive accommodative esotropia. AB - PURPOSE: To evaluate the efficacy of excimer laser refractive surgery as an alternative for optical correction in patients affected by fully refractive accommodative strabismus. SETTING: Eye Clinic, University of L'Aquila, L'Aquila, Italy. METHODS: After a simulation of the cycloplegic correction with contact lenses over a 30-day period, 18 patients (6 men, 12 women, mean age 32.4 years +/ 9.4 [SD]) affected by fully refractive accommodative esotropia had refractive surgery using an excimer laser; 8 patients had photorefractive keratectomy (PRK), and 10 patients had laser in situ keratomileusis (LASIK). RESULTS: The correction of the refractive error with excimer laser allowed a reduction of the angle of deviation in all but 1 patient, who presented with a regression of refractive error and of the angle of deviation 2 years posttreatment. The 2-year follow-up showed that the mean angle of deviation in PRK was 2(Delta) esophoria at near and 0.4(Delta) esophoria at distance (P<.06); in LASIK, it was 1.7(Delta) esophoria at near and 0.2(Delta) esophoria at distance (P<.06). The difference between the 2 groups was not statistically significant at near (P = .56), at distance (P = .74), or for spherical equivalent (P = .16). CONCLUSION: Excimer laser refractive surgery seems to be useful in the correction of fully refractive accommodative esotropia. PMID- 16338559 TI - Biometry of phakic intraocular lens using Scheimpflug photography. AB - PURPOSE: To examine lateral and axial positioning of phakic intraocular lenses (IOLs) with iris fixation in the anterior chamber and to examine short-term stability of the IOL position. SETTING: The Netherlands Opthalmic Research Institute, Amsterdam, the Netherlands. METHODS: Thirty patients participated in the study. Thirty-one eyes were implanted with the 204 type myopia IOL, 14 eyes with the 206 myopia IOL, and 8 eyes with the 203 hyperopia IOL. Scheimpflug slitlamp photographs were made through the optical axis along 4 meridians of the eyes. Ray tracing was used to obtain the lateral and axial position of the IOLs. RESULTS: Centration of the IOL with respect to the pupil's center and the tilt angle of the IOL with respect to the optical axis of the eye were measured. Standard deviation of decentration was 0.21 mm vertically and 0.16 mm horizontally. Standard deviation of tilt was 1.30 degrees vertically and 0.90 degrees horizontally. Tilt and decentration are proportional to each other. Vaulting, the distance between the crystalline lens and the IOL, was constant over a period of 24 months, ranging from 0.2 to 0.8 mm, depending primarily on the radius of curvature of the crystalline lens. A geometric model for this dependence was formulated. CONCLUSION: Phakic IOLs with iris fixation can be positioned in the eye with submillimeter precision. Axial position of iris fixated phakic IOLs over time is excellent. Axial position and vaulting can be predicted when the radius of curvature of the crystalline lens is known. The IOL behaves as if mounted slightly above a sphere-the anterior surface of the crystalline lens. PMID- 16338560 TI - Capsular bag shrinkage after implantation of a capsular bending or capsular tension ring. AB - PURPOSE: To evaluate the influence of a capsular bending ring (CBR) or tension ring (CTR) on capsular bag shrinkage. SETTING: Department of Ophthalmology, Johannes Gutenberg-University, and Department of Medical Biometry, Epidemiology and Informatics, University of Mainz, Mainz, Germany. METHODS: In 92 eyes of 92 patients, a capsular measuring ring (CMR) was implanted after phacoemulsification to measure capsular bag size in vivo. Patients were randomized into 3 groups: The first received a CMR and CBR, the second received a CMR and CTR, and the third received a CMR alone. Measurements were performed preoperatively, intraoperatively, during the first 3 postoperative days, and after 1 and 3 months. Preoperative biometric characteristics such as axial length (AL) and anterior chamber depth were determined. Primary clinical end points were capsular bag size and capsulorhexis diameter measured intraoperatively and by means of slitlamp biomicroscopy and retroillumination photography postoperatively. RESULTS: There were no clinically relevant group differences in AL, anterior chamber depth, or capsulorhexis diameter. Eyes implanted with the CBR showed shrinkage of the capsular bag from 10.6 to 10.4 mm after a median of 3 months (sign test; P = .023); eyes with a CTR showed comparable median capsular bag shrinkage from 10.5 to 10.2 mm (P<.001), whereas eyes without a CTR showed a median shrinkage from 10.5 to 10.0 mm (P<.001). Intraindividual shrinkage was of a similar, although significantly different, order among these groups (median shrinkage 0.15 versus 0.23 and 0.38 mm, respectively; all pairwise Wilcoxon test; P = .050, P = .020, P<.001, respectively). CONCLUSIONS: Capsular shrinkage can be inhibited by a CBR and, to lesser extent, a CTR; the gain in shrinkage prevention is limited when compared with a CMR, however. This conclusion can be drawn only for the specific CBR or CTR and hydrophobic acrylic intraocular lens (IOL) used in this study. Reduction of capsular bag shrinkage after CTR implantation may reduce IOL dislocation and tilt and help to maintain postoperative visual acuity. PMID- 16338561 TI - Simultaneous laser in situ keratomileusis on the stromal bed and undersurface of the flap in eyes with high myopia and thin corneas. AB - PURPOSE: To evaluate simultaneous laser in situ keratomileusis (LASIK) on the stromal bed and the undersurface of the flap in eyes with high myopia and insufficient corneal thickness for conventional LASIK ablation. SETTING: 21 Century Eye Hospital, Seoul Paik Hospital, Inje University, Seoul, Korea. METHODS: Using intraoperative pachymetry, the residual stromal bed thickness was evaluated and simultaneous LASIK was performed on the stromal bed and undersurface of the flap without compromising the posterior 300 mum of the stromal bed. In a prospective series, outcomes in 43 eyes of 27 patients with mean spherical equivalent refraction ranging from -12.5 to -3.75 diopters were analyzed 1, 3, and 6 months after surgery. RESULTS: Postoperative uncorrected visual acuity was 20/30 or better in 35 eyes (81.4%) at 1 month, 36 eyes (83.7%) at 3 months, and 34 eyes (79.1%) at 6 months. During surgery and follow-up, no vision-threatening complications occurred. CONCLUSION: Simultaneous LASIK appears to be a useful surgical aid in LASIK treatment in eyes with high myopia with or without astigmatism when an adequate residual stromal bed does not exist. PMID- 16338562 TI - Effect of cytochrome c peroxidase on corneal epithelial healing process after photorefractive keratectomy. AB - PURPOSE: To evaluate the role of commercially prepared cytochrome c peroxidase eyedrops in corneal epithelial healing after photorefractive keratectomy (PRK). SETTING: Department of Pathophysiological Optics, Faculty of Medicine, University of Bologna, Bologna, Italy. METHODS: Seventy-two eyes of 36 patients affected by low to moderate refractive error (myopia and myopic astigmatism) had uneventful bilateral photorefractive keratectomy (PRK). In each patient, 1 eye (32 eyes) received standard postoperative therapy plus cytochrome c peroxidase eyedrops (3 times a day for 1 week or until corneal reepithelialization was completed, corresponding to 15 000). The fellow eye served as the control and received standard postoperative therapy plus placebo. Patients were monitored daily starting the day after surgery for 7 days to evaluate the corneal reepithelialization rate using a video slitlamp camera with a cobalt blue light. Mean diameter of corneal wounds was measured. Videotaped images were recorded and analyzed by computer planimetry. RESULTS: All the eyes treated with cytochrome c peroxidase eyedrops healed completely before day 5 postsurgery, with a mean reepithelialization time of 91 hours +/- 14 (SD); the mean reepithelialization time was 154 +/- 9 in eyes receiving placebo (P<.05); the mean reepithelialization rate was 0.066 +/- 0.007 mm/hour in the cytochrome c peroxidase group and 0.039 +/- 0.006 mm/hour in the control group (P<.05). There were no statistically significant differences between groups in corneal haze presentation during follow-up (P =.70), perhaps because the time period was too brief (7 days). However, corneal clarity, on slitlamp biomicroscopy in the study group was greater than in the control group. No side effects or toxic effects were documented. CONCLUSIONS: These data suggest that cytochrome c peroxidase significantly accelerates epithelial healing after PRK. Further clinical study should be performed to prove the results obtained in this pilot study and the long-term efficacy of cytochrome c peroxidase to prevent corneal haze. PMID- 16338563 TI - Severe corneal epithelial sloughing during laser in situ keratomileusis as a presenting sign for silent epithelial basement membrane dystrophy. AB - PURPOSE: To report the occurrence of large intraoperative epithelial sloughing during laser in situ keratomileusis (LASIK) as a first diagnostic sign for silent epithelial basement membrane dystrophy (EBMD). SETTING: Refractive Surgery and Cornea Unit, Alicante Institute of Ophthalmology, Alicante, Spain. METHODS: In this retrospective case series, the medical records of all patients with large corneal epithelial sloughing/defects during LASIK from January 1995 to December 2004 were reviewed. All patients who presented normal corneas before LASIK and EBMD changes after LASIK were included in this study. The surgical procedures and postoperative course were recorded. The follow-up period was 12 months for all patients. RESULTS: Eleven eyes of 6 patients were included. The mean age was 35 years +/- 6.1 (SD). Mean preoperative uncorrected visual acuity (UCVA) was 20/500 (0.04 +/- 0.02); 12 months after surgery, mean UCVA increased to 20/27 (0.74 +/- 0.21). There was a significant decrease in best spectacle-corrected visual acuity 1 month after surgery. One year postoperatively, 10 of 11 eyes (91%) were within +/-1.00 diopter of emmetropia. Postoperatively, clinical manifestations of EBMD were observed in all eyes and complications were common. In the early postoperative period, diffuse lamellar keratitis was observed in 6 of 11 eyes (54.5%) and flap microfolds were noted in 2 of 11 (18.2%). One year after surgery, epithelial ingrowth was present in 8 of 11 eyes (72.7%) and flap melting was noted in 4 of 11 (36.4%). CONCLUSION: Occurrence of large intraoperative epithelium sloughing/defects during LASIK might be a diagnostic sign for subclinical EBMD. These patients are predisposed to multiple postoperative complications. Because of the high risk for epithelial sloughing in the second eye, LASIK should not be performed. PMID- 16338564 TI - Pupil decentration and iris tilting detected by Orbscan: anatomic variations among healthy subjects and influence on outcomes of laser refractive surgeries. AB - PURPOSE: To investigate the anatomic variations of entrance pupil decentration and tilting angle of the iris in healthy subjects and the influence of these factors on the outcome of laser in situ keratomileusis (LASIK). SETTING: Minamiaoyama Eye Clinic, Tokyo, Japan. METHODS: The degree of pupil decentration and tilting angle of the iris in 2280 eyes of 1144 myopic patients without abnormal findings by ophthalmologic examination were assessed using Orbscan. Of these, 901 eyes of 467 patients had LASIK. Multiple analysis of variance (ANOVA) was used to determine risk factors for reduction of postoperative best spectacle corrected visual acuity (BSCVA) considering patient age, refractive power, tilting angle of the iris, pupil decentration, and corneal power. RESULTS: The mean pupil decentration in all eyes was 0.19 mm +/- 0.11 (SD) (range 0 to 0.9 mm); tilting angle of the iris was 4.06 +/- 1.41 degrees (range 0.19 to 12.69 degrees). By multiple ANOVA, refractive power, pupil decentration, and tilting angle of the iris were significant for the reduction of BSCVA. CONCLUSIONS: Some eyes with pupil decentration or tilting angle of the iris could not be detected under typical ophthalmologic examination but only with topographic examination. Attention should be paid to eyes with large pupil decentration and tilting angle of the iris because these may be risk factors for reduction of postoperative BSCVA during corneal refractive surgeries. PMID- 16338565 TI - Functional outcomes and patient satisfaction after laser in situ keratomileusis for correction of myopia. AB - PURPOSE: To determine subjective patient satisfaction and self-perceived quality of vision after laser in situ keratomileusis (LASIK) to correct myopia and myopic astigmatism. SETTING: Department of Ophthalmology, Academic Hospital Maastricht, Maastricht, The Netherlands. METHODS: A validated questionnaire consisting of 66 items was self-administered by 142 consecutive patients. Seven scales covering a specific aspect of quality of vision were formulated. Aspects included global satisfaction, quality of uncorrected and corrected vision, quality of night vision, glare, daytime driving, and night driving. Main outcome measures were responses to individual questions and scale scores, and correlations with clinical parameters including refractive outcome, uncorrected visual acuity, best corrected visual acuity, ablation depth, and scotopic pupil-optical zone disparity were obtained. RESULTS: The mean score for the overall satisfaction was 4.1 +/- 0.71 (SD) (scale 0 to 5.0). A total of 92.2% of patients were satisfied or very satisfied with their surgery, 93.6% considered their main goal of surgery achieved, and 92.3% would choose to have LASIK surgery again. Satisfaction with uncorrected vision was 3.03 +/- 0.71. The mean score for glare was 3.0 +/- 0.9. At night, glare from lights was believed to be more important than before surgery by 47.2%. Glare from oncoming car headlights after surgery was reported by 58.4% and was believed to be more bothersome for night driving than before surgery by 52.8%. Night driving was rated more difficult than before surgery by 39.4%, whereas 59.3% had less difficulty driving at night. There was a significant correlation between the uncorrected vision score and the postoperative spherical equivalent (r = 0.245) and postoperative astigmatism (r = 0.265). There was no correlation between the glare or night vision scores and the degree of correction, the amount of ablation depth, or the disparity between the scotopic pupil and the optical zone. CONCLUSIONS: Self-perceived uncorrected vision after LASIK surgery for the correction of myopia and myopic astigmatism appears to be very good and is related to the postoperative residual error. Although the majority of patients postoperatively experienced glare, particularly with driving at night, this was not related to the pupil-optical zone disparity or degree of correction. PMID- 16338566 TI - Quality-of-life improvements in cataract patients with bilateral blue light filtering intraocular lenses: clinical trial. AB - PURPOSE: To compare change in patient-reported vision-related and health-related functioning and quality of life (HRQOL) following bilateral implantation with a new blue light-filtering intraocular lens (IOL) with the results of a similar IOL that does not filter blue light. SETTING: Six clinical sites in the United States. METHODS: Patients were from 6 clinical sites in the United States that performed a high volume of cataract surgeries. The HRQOL assessments occurred via telephone while patients were at home. Patients requiring bilateral cataract extraction were randomly assigned to 1 of the 2 IOL groups for the first eye. The second eye was later implanted with the same type of IOL. Patients and HRQOL data collectors were treatment-masked, but investigators could not be. Health-related functioning and quality of life was measured with the 39-item National Eye Institute Visual Functioning Questionnaire (NEI VFQ-39) and 12-item Short Form Health Survey (SF-12). Assessments were at baseline before implantation in the first eye and 30 to 60 days and 120 to 180 days after implantation of the lens in the second eye. RESULTS: Both IOL types improved most aspects of patients' HRQOL including color vision and driving. The largest gains occurred on the VFQ composite, general vision, near activities, distance activities, driving, mental health, peripheral vision, and role difficulties scales. Significant gains also occurred on color vision and other vision-specific scales as well as the SF-12 physical component summary score. There were no significant differences in HRQOL gains between the IOLs. CONCLUSION: The blue light-filtering IOL improved color vision, driving, and other aspects of HRQOL in a manner similar to that of a lens that does not filter blue light. PMID- 16338567 TI - Effect of postoperative refractive error on visual acuity and patient satisfaction after implantation of the Array multifocal intraocular lens. AB - PURPOSE: To determine the ideal target refraction to optimize visual acuity (VA) and patient satisfaction after implantation of Array SA40N multifocal intraocular lens (IOL) (AMO). SETTING: Inha University College of Medicine, Incheon, Korea. METHODS: The study prospectively enrolled 188 eyes of 163 patients and assigned them to 1 of 3 groups according to their postoperative refractive status: Group 1 (43 myopic eyes, -0.50 diopter [D] to -1.50 D), Group 2 (114 emmetropic eyes, 0.50 D to +0.50 D) and Group 3 (31 hyperopic eyes, +0.50 D to +1.50 D). Uncorrected distance visual acuity (UCDVA), uncorrected near visual acuity (UCNVA), UCDVA under glare conditions, contrast sensitivity, and patient satisfaction were then compared between the 3 groups. RESULTS: Three months postoperatively, UCDVA, UCNVA, UCDVA under glare conditions, and contrast sensitivity were good in all eyes and more than 72% of patients never wore glasses for near vision. In Group 2, UCDVA was significantly better (0.74 +/- 0.20; P<.05) compared with Groups 1 and 3 (0.40 +/- 0.24 and 0.38 +/- 0.30, respectively); UCNVA was also significantly better in Group 2 (0.68 +/- 0.08; P<.05) than in Groups 1 and 3 (0.45 +/- 0.21 and 0.41 +/- 0.17, respectively). However, there were no significant differences in patient satisfaction, contrast sensitivity, and UCDVA under glare conditions between the 3 groups (P>.05). CONCLUSIONS: Aiming for emmetropia rather than myopia when calculating the power for the multifocal intraocular lens may improve visual acuity. However, patients must be considered on an individual basis to meet their expectations and requirements. PMID- 16338568 TI - Effect of preoperative counseling on patient fear from the visual experience during phacoemulsification under topical anesthesia: Multicenter randomized clinical trial. AB - PURPOSE: To determine whether preoperative counseling about potential intraoperative visual experience during phacoemulsification under topical anesthesia reduces fear in patients having cataract surgery. SETTING: The Eye Institute at Tan Tock Seng Hospital, Singapore, Royal Hull Hospitals NHS Trust, Kingston-upon-Hull, and Manchester Royal Eye Hospital, Manchester, United Kingdom. METHODS: In this prospective multicenter randomized clinical trial, patients with cataracts having elective phacoemulsification under topical anesthesia were recruited and randomized into 2 groups. Both groups received routine preoperative counseling regarding risks and benefits of cataract surgery. One group received additional counseling on the potential intraoperative visual experience during phacoemulsification; the other group did not. The patients were then interviewed within 24 hours following phacoemulsification regarding their intraoperative experience. RESULTS: Two hundred nineteen patients were recruited over an 11-month period. There were 104 men and 115 women. The mean age was 68 years (range 20 to 89 years). There were 188 Singaporeans, comprising 168 Chinese, 13 Malays, and 7 Indians, and 31 British patients, all of whom were white. The mean fear score was 0.3 in the group that received additional counseling and 0.9 in the group that did not receive additional counseling (P = .036). The effect of counseling on fear was significant (P = .002) even after controlling for sex, age, and whether first or second cataract surgery. CONCLUSIONS: Preoperative counseling about the potential intraoperative visual experience during phacoemulsification under topical anesthesia helped to reduce the fear from the visual sensations in patients having cataract surgery. PMID- 16338569 TI - Refractive expectations of patients having cataract surgery. AB - PURPOSE: To assess the refractive expectations of patients having elective cataract surgery. SETTING: Queen's Medical Centre, Nottingham, King's Mill Hospital, Sutton-in-Ashfield, and Kidderminster District Hospital, United Kingdom. METHODS: A prospective questionnaire study of patients attending a preoperative assessment clinic for elective cataract surgery was performed. Only first-eye cataract surgery patients were included. Patients were excluded if they had visually impairing ocular comorbidity or if they were mentally unable to complete the questionnaire. RESULTS: One hundred eighty-nine questionnaires were received. Mean patient age was 74 years (range 41 to 97 years). Sixty-four percent of respondents were women, and 90% were retired. On 10-point Likert scales (0 lowest, 10 highest), median patient scores for the perceived likelihood of needing spectacles after surgery were 8 for both distance and near correction. Patients already wearing distance correction thought it significantly more likely that they would need distance glasses postoperatively than those who did not (median likelihood scores 9 and 1, respectively; P<.0001). Similar differences in expectations were demonstrated for near correction. Median score of the importance of not needing spectacles was 8 for both distance and near. Men scored this higher than women, but only for distance. There was a weak negative correlation between the importance of spectacle independence and patient age. CONCLUSIONS: Patients who already wear spectacles expect to need them after cataract surgery. Those not already wearing spectacles do not expect to need them. This latter group is at particular risk for refractive disappointment and complaint. In general, patients consider the opportunity to be free of glasses as very important. PMID- 16338570 TI - Volume-based characterization of postocclusion surge. AB - PURPOSE: To propose an alternative method to characterize postocclusion surge using a collapsible artificial anterior chamber to replace the currently used rigid anterior chamber model. SETTING: Fundacion Oftamologica Los Andes, Santiago, Chile. METHODS: The distal end of a phacoemulsification handpiece was placed inside a compliant artificial anterior chamber. Digital recordings of chamber pressure, chamber volume, inflow, and outflow were performed during occlusion break of the phacoemulsification tip. The occlusion break profile of 2 different consoles was compared. RESULTS: Occlusion break while using a rigid anterior chamber model produced a simultaneous increase of chamber inflow and outflow. In the rigid chamber model, pressure decreased sharply, reaching negative pressure values. Alternatively, with the collapsible chamber model, a delay was observed in the inflow that occurs to compensate the outflow surge. Also, the chamber pressure drop was smaller in magnitude, never undershooting below atmospheric pressure into negative values. Using 500 mm Hg as vacuum limit, the Infiniti System (Alcon) performed better that the Legacy (Alcon), showing an 18% reduction in peak volume variation. CONCLUSIONS: The collapsible anterior chamber model provides a more realistic representation of the postocclusion surge events that occur in the real eye during cataract surgery. Peak volume fluctuation (mL), half volume recovery time(s), and volume fluctuation integral value (mL x s) are proposed as realistic indicators to characterize the postocclusion surge performance. These indicators show that the Infiniti System has a better postocclusion surge behavior than the Legacy System. PMID- 16338571 TI - Adherence of triamcinolone acetonide to various intraocular lens materials. AB - PURPOSE: To determine the adherence of triamcinolone acetonide to various intraocular lens (IOL) materials. SETTING: Department of Ophthalmology, Dokuz Eylul University, Izmir, Turkey. METHODS: Four IOLs of 4 materials (single-piece poly(methyl methacrylate) [PMMA], 3-piece foldable silicone, 3-piece hydrophobic acrylic, and single-piece hydrophilic acrylic) were first immersed in triamcinolone acetonide (40 mg/mL) for 15 minutes and then bathed in a balanced salt solution for another 15 minutes. Afterward, each lens optic was examined under light microscopy and digital images were obtained with a digital color video camera. The percentage of optic area covered with triamcinolone acetonide was determined by image analysis. RESULTS: Mean triamcinolone coverage was 7.62% +/- 4.06% (SD) for PMMA IOLs (range 1.97% to 11.43%), 9.09% +/- 4.60% for silicone IOLs (range 4.70% to 15.32%), 8.75 +/- 7.20% for hydrophobic acrylic IOLs (range 1.31% to 16.86%), and 23.16 +/- 8.53% for hydrophilic acrylic IOLs (range 15.02% to 35.12%). Adherence of triamcinolone acetonide to hydrophilic acrylic lenses was statistically significant when compared with other IOL materials. CONCLUSION: Intraoperative triamcinolone injection may obscure visualization intraoperatively and postoperatively, especially in eyes with hydrophilic acrylic lenses, because triamcinolone acetonide seems to have a tendency to adhere to hydrophilic acrylic lenses in vitro. PMID- 16338572 TI - Treatment-induced shifts of ocular reference axes used for measurement centration. AB - PURPOSE: To determine the shifts of the main corneal reference points in dependence of the chosen centration axis for the treatment. SETTING: Federal Institute of Technology Zurich, Institute of Biomedical Engineering, Zurich, Switzerland. METHODS: Computer simulations were performed on several variants of the Gullstrand-Emsley schematic eye, which was modified by an off-axis fovea. Refractive corrections were simulated by centering Munnerlyn's formula on each of the 4 corneal reference points determined in the preoperative eye: the optical axis, the line of sight, the visual axis, and the first corneal reflex. Subsequently, the postoperative locations of these axes were determined and compared with the preoperative values. RESULTS: The postoperative line of sight was found to depend least on the choice of the preoperative centration axis for both myopic and hyperopic treatments. It undergoes a maximum movement of 0.040 mm when centering a +5 diopter correction on the preoperative line of sight, whereas the corneal reflex, which is used for centering most topography systems, can move by more than 0.10 mm. CONCLUSIONS: Centration of the correction on the preoperative line of sight enabled good comparability between preoperative and postoperative measurements that use the line of sight as a reference axis. Yet, centration of the treatment on the preoperative line of sight does not ensure comparability between preoperative and postoperative measurements that use the corneal reflex as a reference axis such as most corneal topography systems. Axis shifts might lead to misinterpretation of data such as a wrong diagnosis of a decentered ablation or changes in the Zernike representation. PMID- 16338573 TI - Safety and efficacy of levofloxacin 1.5% eyedrops in nonhuman primates having penetrating keratoplasty: clinical and laboratory findings. AB - PURPOSE: To determine the safety and efficacy of levofloxacin 1.5% eyedrops. SETTING: Tulane University Delta Research Primate Center, New Orleans, Louisiana, USA. METHODS: Ten monkeys had preoperative slitlamp examinations, photographs, and confocal microscopy. Levofloxacin 1.5% or control eyedrops were given 4 times a day in both eyes for 2 days and every 5 minutes for the hour before surgery and 4 times a day in both eyes for 1 week. Confocal microscopy, slitlamp examination, and slitlamp photography were repeated preoperatively. Intraoperative aqueous humor and corneal biopsies were taken. RESULTS: There were no differences between groups in slitlamp examinations, slitlamp photography, confocal microscopy, or histology; the mean levofloxacin level in the aqueous humor was 4.49 mug/mL +/ 1.59 SD and in the corneal biopsies, 103.85 +/- 51.48 mug/g. CONCLUSION: Levofloxacin 1.5% drops given on a typical perioperative schedule to monkeys having penetrating keratoplasty showed no clinical or histological signs of toxicity and reached corneal issue levels well over the minimum inhibitory concentrations for all known ocular pathogens. PMID- 16338574 TI - Ophthalmology and vision science research. Part 1: Understanding and using journal impact factors and citation indices. AB - In an increasingly "publish or perish" clinical and academic environment, all clinicians and clinician-scientists involved in research must have a firm understanding of the measures commonly used to assess the quality of scientific journals and, by default, those extended to grade individual articles and authors. The publication of research is a vital part of clinical and experimental research, and citation analyses of research publications have increasingly been adopted as a means of assessing the apparent quality of journals and the research published therein. In the first of a series of articles for those embarking on ophthalmic and vision science research, this paper discusses the key features of citation analysis, concentrating on the 2004 Journal Citation Report figures for the field of ophthalmology that include 42 ophthalmology, vision science, physiological optics, and optometry journals. The Institute for Scientific Information (ISI) calculates a number of parameters including citation counts, Journal Impact Factor (JIF), Immediacy Index, and cited/citing half-life. This article discusses the methods of calculation and possible uses along with current controversies and potential abuses. The JIF and its relevance, potential bias, and limitations are discussed in depth as it has become the most widely used analysis of journal quality. The possible alternatives to ISI citation analysis are presented, and we conclude that citation analysis can be considered a reasonable measure of journal research quality only if used correctly. PMID- 16338575 TI - ASCRS White Paper: Management of infectious keratitis following laser in situ keratomileusis. PMID- 16338576 TI - Nocardia keratitis after laser in situ keratomileusis: clinicopathologic correlation. AB - A case of Nocardia asteroides keratitis occurring 3 weeks after laser in situ keratomileusis (LASIK) in a nontraumatized eye is reported. The patient presented with decreased vision, inflammation, and stromal melting of the LASIK flap, discrete infiltrates, and an anterior chamber cellular reaction. Cultures for acid-fast bacteria grew Nocardia asteroides after 5 days. Infection progressed despite treatment with topical antibiotics and eventually required penetrating keratoplasty (PKP). Postoperatively, the patient was placed on moxifloxacin, a fourth-generation flouroquinolone. The patient experienced a recurrence of Nocardia keratitis at the graft-host interface 2 months after the PKP. This eventually resolved with a combination of topical moxifloxacin and imipenem therapy. PMID- 16338577 TI - Flap tearing during lift-flap laser in situ keratomileusis retreatment. AB - A flap tear occurred during laser in situ keratomileusis (LASIK) retreatment using a flap-lifting technique in 1 eye of 2 patients 4 to 5 months after the primary procedure. In the first case, the tear occurred in a decentered, standard thickness flap (168 mum) in a location close to the corneal limbus and limbal vessels. In the second case, the tear occurred in a well-centered thin flap (116 mum) that involved a peripheral corneal pannus. The false track was identified early, and central extension of the tear was averted. After the flap was successfully dissected, retreatment was performed without further complications. This report suggests that flaps with margins near the limbus or a corneal pannus may be prone to an earlier and stronger healing process at the edge that may lead to a flap tear during LASIK retreatment. This may be of increasing importance because of the trend toward larger flap diameters. PMID- 16338578 TI - Postoperative low-grade endophthalmitis caused by biofilm-producing coccus bacteria attached to posterior surface of intraocular lens. AB - We report a case of low-grade inflammation that developed in the anterior segment after cataract extraction with intraocular lens (IOL) implantation. The 57-year old patient complained of blurred vision 2 weeks after phacoemulsification and IOL implantation in the right eye. Slitlamp biomicroscopy showed many nonpigmented keratoprecipitates. After antibiotic therapy failed, the IOL was removed and aqueous collected. Scanning electron microscopy of the IOL demonstrated many biofilm-producing cocci with slime on the IOL, and aqueous smears showed gram-positive cocci. Two weeks after removal of the IOL, the inflammation disappeared. PMID- 16338579 TI - Salzmann's-like nodular degeneration following laser in situ keratomileusis. AB - We report the development of bilateral Salzmann's-like corneal lesions located at the site of laser in situ keratomileusis (LASIK) flap margins in 2 patients. These lesions resulted in ocular surface irritation as well as induction of irregular astigmatism and associated loss of best corrected visual acuity. Both patients had bilateral LASIK before the occurrence of the corneal lesions. A biopsy was performed on 1 nodule. The epithelium overlying this raised bluish white corneal lesion appeared irregular in thickness, with replacement of Bowman's layer by periodic acid-Schiff positive thickened basement membrane-like material. Underlying this basement membrane was a layer of relatively regular, hypocellular, collagenlike connective tissue, which demonstrated hyalinization on trichrome staining. Based on these 2 cases, it seems possible that the hyposecretion of tears, decreased blink rate, and Dellen effect that often occur following LASIK could produce the corneal irritation needed to induce Salzmann's nodular degeneration. PMID- 16338580 TI - Mitomycin-C assisted photorefractive keratectomy in the treatment of buttonholed laser in situ keratomileusis flaps associated with epithelial ingrowth. AB - The prophylactic intraoperative use of mitomycin-C (MMC) to prevent haze and scarring after excimer laser surface ablation (phototherapeutic/photorefractive keratectomy [PTK/PRK]) in an eye with a previous laser in situ keratomileusis (LASIK) flap buttonhole with epithelial ingrowth is described. A well-centered buttonhole measuring 2.0 mm in diameter was cut within a thin LASIK flap in an amblyopic eye. Over the next 8 weeks, corneal haze and progressive epithelial ingrowth formed centrally. An early transepithelial PTK/PRK approach was chosen to manage the buttonhole together with the epithelial ingrowth and to treat ametropia before the onset of scarring. The approach included epithelial removal with PTK, application of MMC 0.02% for 1 minute, irrigation, a short waiting period to allow for diffusion, PRK correction of -4.0 diopters without nomogram adjustment, and bandage contact lens. A regimen of prednisolone acetate 1% and ofloxacin 0.03% 5 times a day for 1 week (steroid tapered) was prescribed. Epithelial ingrowth was removed successfully. Minimal haze formation was visible 2 weeks after the retreatment but did not reduce best spectacle-corrected visual acuity (BSCVA) and resolved within the next few weeks. After 6 weeks, uncorrected visual acuity was equal to BSCVA preoperatively (20/50). There was no evidence of recurrent epithelial ingrowth or central scarring after 24 months. Transepithelial PTK/PRK was effective in managing central epithelial ingrowth in a buttonholed LASIK flap. Prophylactic intraoperative use of MMC may reduce haze formation and corneal scarring in early treatments and may also prevent recurrent epithelial ingrowth. This approach may offer faster visual recovery and no risk for a repeated buttonhole creation compared with the widespread recutting a new flap after a couple of months. The optimal application time and concentration of MMC need to be established. PMID- 16338581 TI - High IOP as a cause of sudden increased myopia after LASIK. PMID- 16338582 TI - Reduction of subconjunctival hemorrhage with sub-Tenon's anesthesia. PMID- 16338583 TI - Residual cleaner as a cause of intraocular lens slippage. PMID- 16338584 TI - Zoospore encystment and pathogenicity of Phytophthora and Pythium species on plant roots. AB - Seven plant species (lucerne, maize, oat, sugarbeet, sorghum, tomato, wheat) and 12 Pythium and Phytophthora species were used in a comparative study designed to investigate the effects of plant and oomycete inter-specific variation on zoospore encystment density and pathogenicity. Zoospores showed differential encystment behaviour and they encysted more on dicotyledonous than on monocotyledonous plants. Pythium aphanidermatum, P. deliense, and Phytophthora nicotianae were the most aggressive species. Sugarbeet was the most severely attacked plant species followed by tomato while oat plants were relatively unaffected. The relationship between zoospore encystment on roots and disease severity depended on the oomycete-plant combination. Correlation analysis between zoospore encystment density and disease severity indicated low and no significant levels (p.05) of association for most plant-oomycete combinations. PMID- 16338585 TI - Chlamydia trachomatis persistence: an update. AB - Chlamydial persistence is a reversible state generated during conditions deleterious to growth. In persistence, Chlamydia trachomatis remains viable but atypical, with an enlarged, aberrant form and quiescent metabolism. It favours chronic chlamydiosis, leading to serious sequelae. Although the mechanism of persistence formation is still unknown, more reliable molecular approaches tend to confirm that its occurs in vivo, even lasting 3 years. One approach consists of identifying unprocessed rRNA found only in viable Chlamydia, when infection is not apparent. Another approach, referring to the fact that immunity is type specific, consists of showing by genotyping that multiple recurrences are due to the same genovar. At the molecular level, persistence is characterized by increased expression of ct755, one of the three heat shock protein (hsp60)-coding genes. In addition, chromosomal replication occurs continuously, and cell division is blocked possibly due to the repression of genes such as ftsW and amiA. At the immunological level, persistence reveals the failure of host-defence mechanisms because of reduced or suppressed pro-inflammatory or cytotoxic responses. PMID- 16338586 TI - Evaluation of the MicroFoss system for the analysis of Escherichia coli in water. AB - In this study, the performance of the MicroFoss system (Foss, Spain) for the enumeration of Escherichia coli in water samples was evaluated. One hundred and eighty-five samples were analysed both by MicroFoss assay and culture isolation on Tryptone-Bile X-glucuronide agar (TBX), and the correlation coefficient obtained was 0.92. The analysis of 28 new samples using both methods showed a statistically significant relationship at the 99.5% confidence level between log colony forming units obtained by MicroFoss assay and those obtained using growth on TBX agar. Nevertheless, when the level of sample contamination was low, the variability was high. In conclusion, the MicroFoss system is a rapid and simple alternative method for the enumeration of E. coli in water although discordance between the results using these methods in samples with low counts could limit its use for the study of clean water such as potable water. PMID- 16338587 TI - Isolation and molecular characterization of toxigenic Vibrio parahaemolyticus from the Kii Channel, Japan. AB - Studies were conducted on the ecology of potentially pathogenic Vibrio parahaemolyticus in three coastal areas of Kii Channel, Tokushima, Japan. Seawater and seaweed samples were collected seasonally between June 2003 and May 2004. Total and toxigenic strains of V. parahaemolyticus were isolated using most probable number culture and colony blot hybridization. Toxigenic strains were serotyped and further characterized by random amplified polymorphic DNA (RAPD) and ribotyping. Six thousand strains of V. parahaemolyticus were isolated and 18 were found positive for tdh. V. parahaemolyticus were detected in all samples during summer and autumn, and from some samples during winter and spring. Among the toxigenic strains seven serotypes, five ribotypes and RAPD patterns were observed. Seven strains belonged to O3:K6 clone with identical ribotypes and RAPD patterns to that of a pandemic reference strain. The presence of toxigenic V. parahaemolyticus with pandemic potential might indicate a human health risk due to consumption of marine food sources. PMID- 16338588 TI - Synergistic effect of entomogenous fungi on some insecticides against Bihar hairy caterpillar Spilarctia obliqua (Lepidoptera: Arctiidae). AB - A number of fungal parasites infect a wide range of insects and cause epizootics from time to time. Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin are two of the major disease-causing fungi in insects. Investigations were carried out to study the effect of these fungi on the toxicity of endosulfan, imidacloprid, lufenuron, diflubenzuron, dimethoate and oxydemeton methyl against 10-11 days old larvae of Spilarctia obliqua (Walker). For some products the combination treatments showed higher dose mortality response than the sole treatment of fungal conidia or the insecticide. The combination of insecticides with B. bassiana showed 1.26-35.8 fold increase in toxicity of insecticides over sole treatment, while the increase was 1.05-72.0 fold in case of M. anisopliae. Imidacloprid 17.8 SL and oxydemeton methyl 25EC may be used in combination with these fungi for management of S. obliqua. PMID- 16338589 TI - Antibacterial activity directed isolation of compounds from Onosma hispidum. AB - The chemical investigation of the ethanolic extract of the root bark of Onosma hispidum following antibacterial activity directed isolation led to the isolation of 4-hydroxy-3-methoxy cinnamic acid (ferulic acid) and 4-hydroxy-3-methoxy benzoic acid (vanillic acid) which have been reported for the first time in this species. In addition to these compounds, the crude ethanolic extract and methanol fraction exhibited substantial bioactivity against species of corynebacteria, enterococci, staphylococci and streptococci. Ferulic acid was found more bioactive (being relatively more hydrophobic) compared to vanillic acid. PMID- 16338590 TI - Investigation of aminoglycoside modifying enzyme genes in methicillin-resistant staphylococci. AB - Methicillin-resistant staphylococci may also be resistant to some other antibiotics as well as beta-lactams. In this study, co-existence of resistance to methicillin and aminoglycosides was genetically investigated in staphylococci. A total of 50 staphylococci from in-patients, 17 Staphylococcus aureus and 33 coagulase negative staphylococci (CNS) that contained mecA (gene encoding PBP 2a, an altered penicillin-binding protein) determined by polymerase chain reaction (PCR) were included in the study. Aminoglycoside modifying enzyme (AME) genes were investigated using multiplex-PCR. Aminocyclitol-6'-acetyltransferase aminocyclitol-2''-phosphotransferase [aac(6')/aph(2'')] gene (encoding bifunctional acetyltransferases/phosphotransferases) was determined in 66% of the isolates, aminocyclitol-4'-adenylytransferase (ant(4')-Ia) gene (encoding phosphotransferases) in 24%, and aminocyclitol-3'-phosphotransferase (aph(3') IIIa) gene (encoding nucleotidyltransferases) in 8%. Two isolates contained all these three genes. Thirty-six (72%) isolates had at least one of these genes. Three CNS and one S. aureus isolates sensitive to oxacillin had the mecA gene. In conclusion, a high rate of aminoglycoside resistance was determined in methicillin-resistant staphylococci. The aac(6')/aph(2'') was the most frequently detected. PMID- 16338591 TI - Evaluation of blood culture bottles seeded with X-V factors for the detection of Neisseria meningitidis and Haemophilus influenzae. AB - The purpose of this in vitro study was to determine the ability of seeded and not seeded commercial pediatric blood culture bottles to support the growth of the most frequently responsible microorganisms for bacterial meningitides (Neisseria meningitidis, and Haemophilus influenzae). Tests have been carried out with an automated colorimetric pediatric blood culture system, BacTAlert, Organon Teknika. Bottles were inoculated with X-V factors and serial dilutions of the each bacterium in six times (10(1)-10(6) colony forming unit [CFU]/ml). The bottles, which were supplemented with X-V factors, proved to be effective and time to detection (TTD) was shorter than the un-seeded bottles (p0.05). Time difference between seeded and not-seeded bottles was getting greater at high dilutions of both bacteria. We consider that in presence of a few bacteria, the seeding of bottles with X-V factors is very critical obtaining N. meningitidis, and H. influenzae as the causative agents of meningitidis. The recovery rate of the microorganisms, which were isolated from cerebrospinal fluid by using the X-V factor-seeded blood culture bottles, is therefore higher than with the conventional culture methods. PMID- 16338592 TI - Molecular identification of Tuber magnatum ectomycorrhizae in the field. AB - Tuber ectomycorrhizae in a Tuber magnatum "truffiere", located in Central Italy, were studied using molecular methods. Specifically, RFLP-ITS analyses, ITS sequencing and specific probes hybridization were used to identify 335 Tuber-like ectomycorrhizal morphotypes. Molecular identification was possible even when distinct morphological characteristics were lacking. For the first time, T. magnatum ectomycorrhizae and other coexisting Tuber species collected in the field were analysed using molecular tools for unambiguous identification. Although the "truffiere" under investigation yields good harvests of T. magnatum fruiting bodies, the percentage of T. magnatum ectomycorrhizae found was very low (less than 4.4% of the 335 root tips analysed), whereas the percentages of Tuber maculatum and Tuber rufum were considerably higher (48.9% and 19.0%, respectively). PMID- 16338593 TI - Acid response of exponentially growing Escherichia coli K-12. AB - Induction of acid tolerance response (ATR) of exponential-phase Escherichia coli K-12 cells grown and adapted at different conditions was examined. The highest level of protection against pH 2.5 challenges was obtained after adaptation at pH 4.5-4.9 for 60 min. To study the genetic systems, which could be involved in the development of log-phase ATR, we investigated the acid response of E. coli acid resistance (AR) mutants. The activity of the glutamate-dependent system was observed in exponential cells grown at pH 7.0 and acid adapted at pH 4.5 in minimal medium. Importantly, log-phase cells exhibited significant AR when grown in minimal medium pH 7.0 and challenged at pH 2.5 for 2 h without adaptation. This AR required the glutamate-dependent AR system. Acid protection was largely dependent on RpoS in unadapted and adapted cells grown in minimal medium. RpoS dependent oxidative, glutamate and arginine-dependent decarboxylase AR systems were not involved in triggering log-phase ATR in cells grown in rich medium. Cells adapted at pH 4.5 in rich medium showed a higher proton accumulation rate than unadapted cells as determined by proton flux assay. It is clear from our study that highly efficient mechanisms of protection are induced, operate and play the main role during log-phase ATR. PMID- 16338594 TI - Cold-active acid beta-galactosidase activity of isolated psychrophilic basidiomycetous yeast Guehomyces pullulans. AB - In the present study, psychrophilic yeasts, which grow on lactose as a sole carbon source at low temperature and under acidic conditions, were isolated from soil from Hokkaido, Japan. The phenotypes and sequences of 28S rDNA of the isolated strains indicated a taxonomic affiliation to Guehomyces pullulans. The isolated strains were able to grow on lactose at below 5 degrees C, and showed cold-active acid beta-galactosidase activity even at 0 degrees C and pH 4.0 in the extracellular fractions. Moreover, K(m) of beta-galactosidase activity for lactose in the extracellular fraction from strain R1 was found to be 50.5 mM at 10 degrees C, and the activity could hydrolyze lactose in milk at 10 degrees C. The findings in this study indicate the possibility that the isolated strains produce novel acid beta-galactosidases that are able to hydrolyze lactose at low temperature. PMID- 16338595 TI - Effect of the tropical grass Brachiaria brizantha (Hochst. ex A. Rich.) Stapf on microbial population and activity in petroleum-contaminated soil. AB - The effect of the tropical pasture grass Brachiaria brizantha on numbers of bacteria, fungi and degraders of alkanes, aromatics, cycloalkanes and crude oil in petroleum hydrocarbon contaminated and uncontaminated savannah soil was evaluated. Substrate induced soil respiration and soil pH were compared between planted and unplanted soil. B. brizantha had a mostly increasing effect on microbial numbers. As an exception, growth of bacteria was not or negatively affected. Microbial respiration and pH were always lower in planted than in unplanted soil. Low pH may result from enhanced oil degradation in planted soil leading to an accumulation of organic acids. A comparable stimulation of crude oil degraders and fungi in planted soil points to the importance of fungi. Since they tolerate lower pH values than bacteria, they are considered to play a central role in oil degradation. Given that the enhancement of crude oil degradation under the influence of B. brizantha could not clearly be correlated to microbial numbers and activity, other factors like oxygen availability, plant enzymes and synergistic degradation by microbial consortia have to be considered. PMID- 16338596 TI - Correlation between asthma and irritable bowel syndrome in a general population in Iran in 2003. AB - BACKGROUND: Associations have been reported between irritable bowel syndrome (IBS) and asthma. AIMS: To explore the inter-relations between these conditions in a general population sample. SUBJECTS: A randomly selected community sample of 5492 adults. METHODS: All of the subjects were invited for a medical interview. Those subjects suspected to have either asthma and/or IBS underwent spirometry with post-bronchodilator test if indicated. The labeling of subjects as having IBS was based on Rome II criteria. RESULTS: 4762 subjects agreed to take part (86.7% response). Prevalence rates for IBS, and currently active asthma were 7.1% and 3.8%, respectively. Logistic regression showed independent associations between IBS and most asthma symptom categories. There was no significant independent association between IBS and chronic bronchitis. The odds ratio with 95% confidence interval (CI) for IBS and current asthma was 1.79 (1.06-3.03). CONCLUSIONS: These observations indicate the necessity of further community-based studies to elucidate the possible common pathogenic mechanisms involved in two disease entities. PMID- 16338597 TI - Asthma-related resource use and cost by GINA classification of severity in three European countries. AB - BACKGROUND: This study assessed the relationship between asthma burden and asthma severity in France, Italy, and Spain. METHODS: Adult asthmatics, 18-55 years of age, completed a questionnaire while visiting a respiratory physician in 1998 and 1999. Asthma severity was categorized by physicians as intermittent, mild persistent, moderate persistent, or severe persistent according to Global Initiative for Asthma (GINA) guidelines. RESULTS: Totals of 282 patients in France, 500 in Italy, and 296 in Spain entered the study. There were few differences between the three countries in the asthma symptom burden. Most patients with persistent asthma had used inhaled corticosteroids in the previous 14 days. Unexpectedly, 35% (Italy) to 83% (Spain) of patients with intermittent asthma also had used inhaled corticosteroids. In Spain, visits to the emergency department were more frequent (OR 7.0, 95% CI 4.9-10.0 with Italy as reference) and the costs of emergency care in all asthma severity categories were up to 10 times higher than in Italy and France. The frequency of hospitalizations did not differ systematically between the three countries. CONCLUSIONS: Inadequate control of asthma symptoms among patients with severe persistent asthma could not be entirely explained by under-prescribing of asthma medications. The use of inhaled corticosteroids by patients with intermittent asthma might reflect misclassification of asthma severity, possibly due to difficulty in interpreting the GINA guidelines. The relatively high cost of emergency care in Spain does not appear to be related to greater asthma severity or poorer symptom control, but may be a feature of the Spanish health care system. PMID- 16338598 TI - Different breathing patterns in healthy and asthmatic children: responses to an arithmetic task. AB - Asthma patients have been reported to be sensitive to breathlessness, independent of the degree of airway obstruction. Paying attention and task performance may induce changes in breathing pattern and these in turn may mediate such a feeling. The present experiment investigates whether strained breathing induced by an arithmetic task was different in children with asthma compared to healthy children. METHODS: Seven healthy and eight asthmatic but symptom-free school children were equipped with electrodes for surface electromyographic (EMG) measurements of diaphragm, abdominal and intercostal (IC) muscles and with a strain gauge to monitor the pattern of breathing at rest and during an arithmetic task. The relative duration of exhalation and the relative speed of exhalation are used as measures of straining. The phase angle of maximal respiratory muscle activities relative to the maximal chest extension (MCE) are additional discriminating parameters. RESULTS: Asthmatic children breathed more slowly and already at rest the phase of their respiratory muscle activity appears to be different. While in healthy children the maximal activity of the (left)abdominal muscles occurred 5+/-29% later than the MCE, in children with asthma the maximal activity occurred 26+/-30% of the cycle earlier than MCE. In children with asthma the activity of the IC muscles starts weaning already at 10+/-30% before MCE, in contrast to the healthy children in which intercostal muscle weaning starts only at 1+/-24% after MCE. During arithmetic, the significant difference between the groups in this respect disappeared. CONCLUSION: Children with asthma show, even at rest, signs of respiratory muscle straining, probably in order to keep close control over the airflow in a similar way as healthy children during mental tasks. Such a 'careful' breathing pattern may work to prevent airway irritation also when they are free of symptoms. PMID- 16338599 TI - Corticosteroid sparing effects of vitamin C and magnesium in asthma: a randomised trial. AB - AIM: The study aims to assess the a priori hypothesis that regular supplementation with vitamin C or magnesium will permit a reduction in the corticosteroid dose required to maintain asthma control in adults. METHODS: We invited all participants recruited from primary care centres who completed a parallel-group, randomised, placebo-controlled, 16-week supplementation trial of 1g/day vitamin C or 450 mg/day magnesium to continue and participate in a structured corticosteroid reduction protocol over 10 weeks. RESULTS: A total of 92 participants (29 vitamin C, 31 magnesium and 32 placebo) entered the study. Assuming no reduction in corticosteroid dose in the 10 who subsequently withdrew, the geometric mean reductions in inhaled corticosteroid dose achieved with vitamin C, magnesium and placebo were 49, 13 and 11 microg, respectively. Relative to placebo, the unadjusted effect of vitamin C was significant, and remained at borderline significance after adjustment for baseline corticosteroid dose (relative reduction ratio=4.03, 95% CI 0.95 to 17.1, P=0.06). CONCLUSIONS: We conclude that while vitamin C supplements may have modest corticosteroid sparing effects and hence the potential to reduce exposure to their side effects, magnesium supplements have no effect on the inhaled corticosteroid dose required to maintain asthma control. PMID- 16338600 TI - Communication, communication, communication! PMID- 16338601 TI - Adolescent chronic pelvic pain. AB - The presentation of chronic pelvic pain in the adolescent can at times be quite daunting. A careful and insightful approach to obtaining the history and physical examination must be implemented while maintaining an appreciation of the various stages of adolescent development. The etiologies can range from gynecologic to nongynecologic causes. The ability to render an early diagnosis and appropriate treatment in this population of patients can significantly improve future reproductive health outcomes. The following minireview will outline a systematic approach to the adolescent with chronic pelvic pain. PMID- 16338602 TI - Parents and partners: enhancing participation in contraception use. AB - When adolescent girls choose to have sexual intercourse, it is important for them to have the knowledge, resources, and communication skills necessary to avoid unintended pregnancies. There are many barriers to contraceptive use that are specific to adolescents, and it is possible that parents and partners could help to support consistent and correct use of contraception. This paper reviews the existing literature on the role of parents and partners, and discusses the implications of this knowledge base for clinical practice. The specific recommendations in this manuscript are designed to aid health care providers in supporting the involvement of parents and partners in the lives of adolescent girls in order to promote the positive emotional and physical growth of adolescent girls. PMID- 16338604 TI - Attitudes about human papillomavirus vaccine among family physicians. AB - STUDY OBJECTIVE: Human papillomavirus (HPV) vaccines will soon be available for clinical use, and the effectiveness of vaccine delivery programs will depend largely upon whether providers recommend the vaccine. The objectives of this study were to examine family physicians' attitudes about HPV immunization and to identify predictors of intention to recommend immunization. DESIGN: Cross sectional survey instrument assessing provider and practice characteristics, knowledge about HPV, attitudes about HPV vaccination, and intention to administer two hypothetical HPV vaccines. PARTICIPANTS: Surveys were mailed to a national random sample of 1,000 American Academy of Family Physicians (AAFP) members. MAIN OUTCOME MEASURE: Intention to administer two hypothetical HPV vaccines (a cervical cancer/genital wart vaccine and a cervical cancer vaccine) to boys and girls of different ages. RESULTS: One hundred fifty-five surveys (15.5%) were returned and 145 were used in the final sample. Participants reported higher intention to recommend both hypothetical HPV vaccines to girls vs. boys (P < 0.0001) and to older vs. younger adolescents (P < 0.0001). They were more likely to recommend a cervical cancer/genital wart vaccine than a cervical cancer vaccine to boys and girls (P < 0.001). Variables independently associated with intention (P < 0.05) included: female gender of provider, knowledge about HPV, belief that organizations such as the AAFP would endorse vaccination, and fewer perceived barriers to vaccination. CONCLUSIONS: Female gender, knowledge about HPV, and attitudes about vaccination were independently associated with family physicians' intention to recommend HPV vaccines. Vaccination initiatives directed toward family physicians should focus on modifiable predictors of intention to vaccinate, such as HPV knowledge and attitudes about vaccination. PMID- 16338605 TI - Intersex conditions in children and adolescents: surgical, ethical, and legal considerations. AB - INTRODUCTION: Approximately one in 2000 children globally is born with an intersex condition. There is unfortunately a relative paucity of data on the choices and the surgical and psychosocial outcomes in patients who undergo genital surgery for intersex conditions and ambiguous genitalia, especially in developing countries. Specialists in these and other countries, where patient follow-up is generally poor, are faced with the daunting task of offering the appropriate medical and surgical management, in the absence of guidelines or recommendations. SURGICAL CONSIDERATIONS: A surgical procedure in these patients sometimes involves clitoral recession, reduction, vaginoplasty, and gonadectomy. The best surgical outcome is likely to be achieved with a multidisciplinary surgical team; however, the choice of surgery and appropriate timing remains controversial. Some authors have suggested delaying surgery until the child becomes competent to make his/her own decisions. LEGAL/ETHICAL CONSIDERATIONS: All procedures should conform to an ethical code of practice and be in the interest of the child. Exhaustive counseling of all parties and informed consent is of paramount importance, as is adherence to laws that protect the rights of the child as outlined in respective constitutions. RECOMMENDATIONS: Recommendations in this article, which have been put together from the combined input of three departments, are broad-based. They emphasize the need for extensive counseling, informed consent, adherence to ethical and legal norms, a multidisciplinary input and a shift away from a paternalistic approach. PMID- 16338606 TI - Enhancing recruitment and retention of minority young women in community-based clinical research. AB - Women are disproportionately affected by the sexually transmitted infections (STI) epidemic, with African-Americans and Latinos at significantly higher risk for STIs than Caucasians. Successful recruitment and retention strategies used with young minority women in community-based STI prevention or intervention research have not been previously reported. This communication presents eight key strategies learned in the recruitment and retention of 16- to 21-year-old urban women participating in a 12-month randomized clinical trial designed to promote STI screening to decrease the duration of untreated chlamydia and gonorrhea infection. Strategies learned include: (1) Educate clinic staff on the rigors of study design; (2) Facilitate a team effort between clinical and research staff; modify recruitment procedures, as needed; (3) Provide prospective participants the option of enrolling by return appointment; (4) Anticipate a diminishing recruitment pool over time; (5) Set positive recruitment tone at the beginning of each clinic session; (6) Consider participants' mothers as important points of contact; (7) Match communication styles to participant contacts; and (8) Consider a variety of retention techniques. Together, these strategies helped to reinforce participant's commitment to the project, facilitated their attendance at interviews, and encouraged them to adhere to the treatment protocol. PMID- 16338607 TI - What's your diagnosis? A 3-month-old girl with an ulcerated perineal plaque. PMID- 16338608 TI - Absence of the clitoris in a 13-year-old adolescent: medical implications for child and adolescent health. AB - BACKGROUND: Isolated absence of the clitoris is a rare entity with medical and sexual implications for patients. CASE: As part of an examination for alleged sexual abuse, a 13-year-old female was noted to have an apparent absence of the clitoris. Congenital absence of the clitoris was felt to be the most likely diagnosis. The differential diagnosis and medical implications are reviewed. CONCLUSION: Isolated absence of the clitoris is a rare entity. When faced with the finding, a detailed medical evaluation should be performed to clarify the nature of the finding, rule out other potential anomalies and help in the counseling of the patient. PMID- 16338609 TI - Voluminous mucinous cystadenoma of the ovary in a 13-year-old girl. AB - BACKGROUND: Mucinous cystadenoma of the ovary constitutes about 15% of ovarian tumors. It is more common in women between 20 and 40, but is rare in teenagers and exceptional in pre-menstrual girls. Malignant transformation is possible in 5 10% of cases. Spontaneous or iatrogenic breaks are more common. It makes the different diagnosis of pseudomixoma peritonei more difficult. CASE REPORT: A 13 year-old girl came to the National Cancer Institute "Regina Elena" of Rome, with increasing abdominal distension over 2 months. The abdominal mass was removed and a left salpingo-oophorectomy was performed. Histological examination of the mass revealed a 40-cm diameter mucinous cystadenoma of the ovary. CONCLUSION: The rapid enlargement of the ovarian mass, the young age and the difficulty of certain pre-operative diagnosis, make this case interesting. Management of this pathology and fertility-conserving treatment need careful follow-up because of the possibility of recurrence in the remaining ovary. PMID- 16338610 TI - Premenarchal, recurrent vaginal discharge associated with an incomplete obstructing longitudinal vaginal septum. AB - STUDY OBJECTIVE: To describe an unusual, premenarchal presentation of an obstructive vaginal anomaly. DESIGN: Case Report. SETTING: University Medical Center. PARTICIPANTS: Premenarchal subject INTERVENTIONS: Vaginogram, vaginal septum resection. RESULTS: Vaginal septum resection with resolution of vaginal discharge. CONCLUSIONS: This case demonstrates some of the typical features of uterus didelphys bicollis with incomplete obstructing hemivagina, but had a unique presentation with premenarchal, recurrent vaginal discharge. Typically, patients with an obstructing mullerian anomaly present after menarche with pelvic pain and a mass. The vaginogram assists in the preoperative definition of abnormal anatomy which allows the surgeon to develop the most appropriate surgical approach. Resection of this incompletely obstructing vaginal septum resulted in resolution of the recurrent vaginal discharge. PMID- 16338611 TI - Familial occurrence of imperforate hymen. AB - Imperforate hymen is uncommon, occurring in 0.1 % of newborn females. Non syndromic familial occurrence of imperforate hymen is extremely rare and has been reported only three times in the English literature. The authors describe two cases in a family across two generations, one presenting with chronic cyclical abdominal pain and the other acutely. There were no other significant reproductive or systemic abnormalities in either case. Imperforate hymen occurs mostly in a sporadic manner, although rare familial cases do occur. Both the recessive and the dominant modes of transmission have been suggested. However, no genetic markers or mutations have been proven as etiological factors. Evaluating all female relatives of the affected patients at an early age can lead to early diagnosis and treatment in an asymptomatic case. PMID- 16338612 TI - The timing of the first pelvic exam. PMID- 16338614 TI - Use of DMPA by adolescents. PMID- 16338616 TI - National Institutes of Health consensus development project on criteria for clinical trials in chronic graft-versus-host disease: I. Diagnosis and staging working group report. AB - This consensus document is intended to serve 3 functions. First, it standardizes the criteria for diagnosis of chronic graft-versus-host disease (GVHD). Second, it proposes a new clinical scoring system (0-3) that describes the extent and severity of chronic GVHD for each organ or site at any given time, taking functional impact into account. Third, it proposes new guidelines for global assessment of chronic GVHD severity that are based on the number of organs or sites involved and the degree of involvement in affected organs (mild, moderate, or severe). Diagnosis of chronic GVHD requires the presence of at least 1 diagnostic clinical sign of chronic GVHD (e.g., poikiloderma or esophageal web) or the presence of at least 1 distinctive manifestation (e.g., keratoconjunctivitis sicca) confirmed by pertinent biopsy or other relevant tests (e.g., Schirmer test) in the same or another organ. Furthermore, other possible diagnoses for clinical symptoms must be excluded. No time limit is set for the diagnosis of chronic GVHD. The Working Group recognized 2 main categories of GVHD, each with 2 subcategories. The acute GVHD category is defined in the absence of diagnostic or distinctive features of chronic GVHD and includes (1) classic acute GVHD occurring within 100 days after transplantation and (2) persistent, recurrent, or late acute GVHD (features of acute GVHD occurring beyond 100 days, often during withdrawal of immune suppression). The broad category of chronic GVHD includes (1) classic chronic GVHD (without features or characteristics of acute GVHD) and (2) an overlap syndrome in which diagnostic or distinctive features of chronic GVHD and acute GVHD appear together. It is currently recommended that systemic therapy be considered for patients who meet criteria for chronic GVHD of moderate to severe global severity. PMID- 16338617 TI - Cytolytic pathways used by effector cells derived from recipient naive and memory T cells and natural killer cells in resistance to allogeneic hematopoietic cell transplantation. PMID- 16338618 TI - Perspectives on gene therapy for immune deficiencies. PMID- 16338619 TI - High unbound mycophenolic acid concentrations in a hematopoietic cell transplantation patient with sepsis and renal and hepatic dysfunction. PMID- 16338620 TI - A CD4 domain 1 CC' loop peptide analogue enhances engraftment in a murine model of bone marrow transplantation with sublethal conditioning. AB - Host CD4(+) T cells that survive sublethal or even lethal preconditioning regimens can participate in the process of hematopoietic stem cell graft rejection, particularly when the transplantations are performed across a major histocompatibility complex (MHC) class II barrier. To enhance donor marrow engraftment, we tested the efficacy of a small synthetic cyclic heptapeptide, 802 2 (CNSNQIC), which was designed to closely mimic the CD4 domain 1 CC' surface loop, theoretically involved in CD4/MHC class II complex oligomerization and subsequent CD4(+) T-cell activation. Previously, this peptide was found to have inhibitory activity in murine models for CD4(+) T cell-dependent graft-versus host disease and skin allograft rejection. Herein, we used the MHC class II- disparate bm12 --> B6-CD45.1 sublethal irradiation transplantation model to test the possibility that the 802-2 peptide could enhance the engraftment of donor T cell-depleted bone marrow (ATBM). Sublethally irradiated B6-CD45.1 mice that received bm12 ATBM in combination with the 802-2 peptide demonstrated increased donor marrow cell engraftment as compared with mice that received ATBM alone; this suggests that the 802-2 peptide may be useful as an immunomodulating agent to overcome MHC class II mismatch barriers in hematopoietic stem cell transplantation. PMID- 16338621 TI - Differences in characteristics of US hematopoietic stem cell transplantation centers by proportion of racial or ethnic minorities. AB - Racial or ethnic minorities with leukemia who receive HLA-identical sibling hematopoietic stem cell transplants (HSCTs) are reported to have worse survival when compared with whites. Characteristics of US HSCT centers according to the proportion of ethnic minorities who undergo transplantation were compared to explore systematic differences among centers; the association with 100-day mortality was evaluated to determine whether center factors may explain the observed discrepant survival among ethnic minorities. One hundred sixteen US transplantation centers that performed HLA-identical sibling transplantations for leukemia were analyzed. We compared physician and health care provider staffing, transplantation unit procedure and resources, and medical center organization according to the volume procedure ratio of ethnic minorities who underwent transplantation and also according to the ratio of Hispanics who underwent transplantation. Centers that performed transplantation in a higher proportion of ethnic minorities were more likely to perform fewer transplantations per year, to have fewer devoted transplant beds, to be in an urban setting, to have a lower physician to patient volume ratio, and to follow up survivors 1 year after transplantation. Centers that performed transplantation in a higher proportion of Hispanics were more likely to perform fewer transplantations per year and to have fewer devoted transplantation beds, were less likely to perform outpatient transplantations, were more likely to be in an urban setting, and were less likely to have posttransplantation immunization protocols. Observed differences in center factors were not associated with 100-day mortality after adjustment for disease severity. Our results suggest that the inferior survival reported in ethnic minorities after HSCT may not be readily explained by center effects. PMID- 16338622 TI - Favorable outcome for infant acute lymphoblastic leukemia after hematopoietic stem cell transplantation. AB - Infants with acute lymphoblastic leukemia (ALL) have a poor prognosis when treated with standard chemotherapy. A subset of these infants, particularly those with mixed-lineage leukemia (MLL) rearrangements, has a high likelihood of relapse. Hematopoietic stem cell transplantation (HSCT) performed early in first remission may improve outcome. We present the results of 16 patients with infant ALL who were treated with HSCT in first remission. Six patients were < or =6 months of age at diagnosis, 11 had an initial white blood cell count of >50000/microL, and all patients with determinable cytogenetics had a high-risk karyotype [t(4:11) abnormality or other MLL rearrangement]. All patients received 150 cGy of total body irradiation for 8 doses (1200 cGy). Fifteen of 16 patients received etoposide at 1000 mg/m(2) as a continuous infusion over 24 hours and cyclophosphamide at 60 mg/kg/d for 3 days. Eight patients received HSCT from an HLA-identical sibling, and 8, from unrelated cord blood. Twelve (75%) patients remain long-term survivors (median follow-up, 4.7 years). Two patients, 1 of whom had minimal residual disease at HSCT, died after relapse following HSCT. Two patients died of transplant-related causes. The HSCT was well tolerated; 15 patients achieved neutrophil engraftment at a median of 16 days. Acute and chronic graft-versus-host disease were minimal in these patients. These results support the use of HSCT in the treatment of infant ALL, especially when used as consolidation in first remission. The risk of relapse seems to be decreased with this approach. Further work is being performed to determine the long-term effects from this therapy. PMID- 16338623 TI - ABO blood group barrier in allogeneic bone marrow transplantation revisited. AB - Reports have shown a worse outcome for donor-recipient pairs mismatched for ABO blood groups in bone marrow transplantation (BMT). These studies, however, included small and heterogeneous study populations, and not all considered bidirectional ABO incompatibility separately. Because the issue remains controversial, we analyzed the effect of ABO mismatch on the overall survival, transplant-related mortality, and occurrence of acute and chronic graft-versus host disease (GVHD) in a large homogenous group of patients undergoing allogeneic BMT. A total of 3103 patients with early-stage leukemia who underwent transplantation between 1990 and 1998 with bone marrow from an HLA-identical sibling and who were reported to the Center for International Blood and Marrow Transplant Research were studied. The median follow-up was 54 months. A total of 2108 (67.9%) donor-recipient pairs were ABO identical, 451 (14.5%) had a minor mismatch, 430 (13.9%) had a major mismatch, and 114 (3.7%) had a bidirectional ABO mismatch. The groups did not differ significantly in patient or donor characteristics except for more female-to-male sex mismatch in the bidirectional ABO mismatch group (P = .017). In multivariate models of overall survival, transplant-related mortality, and grade II to IV acute GVHD, there were no significant differences among the 4 groups. Bidirectional ABO mismatch was associated with a significantly higher risk of grade III or IV acute GVHD (hazard ratio, 1.869; 95% confidence interval, 1.192-2.93; P = .006). Patients with major ABO mismatch received red blood cell transfusions (P = .001) for a longer timer after transplantation and had a slightly slower neutrophil recovery (P < .001). There was no evidence of a substantial effect of ABO blood group incompatibility on the outcome of conventional BMT among patients with leukemia. PMID- 16338624 TI - Interleukin 12 is associated with reduced relapse without increased incidence of graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. AB - Interleukin (IL)-12 has antitumor effects in murine studies. To evaluate this clinically, we investigated whether high levels of circulating IL-12 in patients after allogeneic hematopoietic stem cell transplantation (HSCT) are associated with improved relapse-free survival. We prospectively studied 134 patients undergoing HSCT. Median follow-up was 1158 days (range, 70-1792 days). Plasma IL 12 levels were measured before transplantation and on days 0, +4, +7, and +14 after transplantation. The highest levels were seen on days +4 and +7 and were categorized by a cluster analysis of the logarithmically transformed IL-12 concentrations, which were then correlated with relapse-free survival. Forty-six patients had low levels of IL-12 (median, 2 pg/mL; range, 0-6.5 pg/mL), 49 patients had medium levels (median, 20.5 pg/mL; range, 7-75.5 pg/mL), and 25 patients had high levels (median, 181 pg/mL; range, 84-623 pg/mL). Patients with high IL-12 levels before transplantation had the highest increase after transplantation. With a multivariate Cox model for relapse onset, with the low IL 12 level as the reference, patients in the high-IL-12 group had an adjusted hazard ratio of 0.27 (95% confidence interval, 0.09-0.79), and medium group patients had a hazard ratio of 0.65 (95% confidence interval, 0.31-1.36). The incidences of relapse at 500 days by Kaplan-Meier analysis by IL-12 group were 23.0% (high group), 40.3% (medium group), and 48.8% (low group). There was no association between IL-12 levels and the risk of acute graft-versus-host disease (GVHD; P = .51) or chronic GVHD (P = .28). In conclusion, high IL-12 levels after HSCT are associated with improved relapse-free survival without increasing the risk for GVHD. Patients with high pretransplantation IL-12 levels have an increased likelihood of higher posttransplantation IL-12 levels, possibly because of a host-graft interaction, and this may predispose to better clinical outcomes. PMID- 16338625 TI - Cytoprotection by amifostine during autologous stem cell transplantation for advanced refractory hematologic malignancies. AB - This study evaluated whether amifostine protects against mucositis and other toxicities in patients with advanced, refractory, or recurrent hematologic malignancies undergoing high-dose chemotherapy and total body irradiation. Thirty five patients (20 with non-Hodgkin lymphoma, 12 with Hodgkin disease, and 3 with acute myelogenous leukemia) who underwent autologous stem cell transplantation were conditioned with total body irradiation 2 Gy twice daily on days -8 through 6; cyclophosphamide 6 g/m(2), etoposide 1.8 g/m(2), and carboplatin 1 g/m(2) on days -5 through -3; and amifostine 500 mg/m(2) on days -8 through -2. Prior institutional experience in patients treated without amifostine was used as a historical comparison (no-amifostine group). Severe mucositis occurred in 14 (40%) of 35 patients in the amifostine group, compared with 33 (94%) of 35 in the no-amifostine group (P < .0001). Total parenteral nutrition was used by 4 (11%) of 35 amifostine-treated patients and 34 (97%) of 35 no-amifostine patients (P < .0001). The median duration of narcotic use decreased from 15.5 days with no amifostine to 11 days with amifostine (P = .002). Granulocyte and platelet engraftment times were similar. Prospective trials with innovative designs and clearly defined stopping rules are warranted to confirm whether amifostine reduces the toxicities of a myelosuppressive conditioning regimen before autologous stem cell transplantation without compromising therapeutic response. PMID- 16338626 TI - Human mesenchymal stem cells and cyclosporin a exert a synergistic suppressive effect on in vitro activation of alloantigen-specific cytotoxic lymphocytes. PMID- 16338627 TI - Tetrahydrobiopterin and maternal PKU. AB - A 29-year-old woman with PKU is presented, who was successfully treated with phenylalanine restriction as well as oral BH4 during this pregnancy, with a normal outcome. Her PAH mutation was R408W/F39L. Remarkably, the blood phenylalanine control was easily accomplished during this pregnancy. The lack of nausea and vomiting during the first trimester suggests that the occurrence of CHD in babies born to women with PKU may be reduced with BH4. PMID- 16338628 TI - 'Reasonable access' to primary care: assessing the role of individual and system characteristics. AB - Access to health care continues to be an important issue for health policy makers, researchers, service providers and consumers alike. In countries with universal health care coverage, services are generally free at the point of delivery which is intended to provide equitable access to care for all residents regardless of their individual situations. Past studies have pointed to the importance of individual characteristics such as age, gender, and socioeconomic status in shaping access to health care but much less is known about the role of health system characteristics. The purpose of this study is to explore, by means of qualitative research methods, the extent to which individual and system factors shape access to primary health care services for residents living in two contrasting neighbourhoods in Hamilton, Ont., Canada. Semi-structured, in-depth interviews were conducted with 40 men and women. The interviews probed participants about their experiences with primary care, barriers to receiving care and their general perceptions of the health care system. The interviews demonstrated the existence of many system-related barriers to receiving health care (e.g., wait times, geographic inaccessibility and quality of care) and some individual-related barriers (e.g., work or family responsibilities). While the findings revealed little difference between the neighbourhoods in terms of accessibility problems and barriers, differences between men and women were evident. PMID- 16338629 TI - Health related migration: evidence of reduced "urban-drift". AB - The aim of this study was to determine if the onset of serious disease triggers a different intra-state migratory response from patterns observed in the healthy population. The analysis was carried out using linked administrative data. The onset of serious disease triggered a reduction in the rate of endocentric migration in remote and rural populations. Urban drift occurred only in people with mental illness in rural locations. Rural and remote communities appear to suffer from an unhealthy selection force, with persons unable to migrate centrally to access services due to the onset of the physical illness they require treatment for. PMID- 16338630 TI - Whose health is affected by income inequality? A multilevel interaction analysis of contemporaneous and lagged effects of state income inequality on individual self-rated health in the United States. AB - The empirical relationship between income inequality and health has been much debated and discussed. Recent reviews suggest that the current evidence is mixed, with the relationship between state income inequality and health in the United States (US) being perhaps the most robust. In this paper, we examine the multilevel interactions between state income inequality, individual poor self rated health, and a range of individual demographic and socioeconomic markers in the US. We use the pooled data from the 1995 and 1997 Current Population Surveys, and the data on state income inequality (represented using Gini coefficient) from the 1990, 1980, and 1970 US Censuses. Utilizing a cross-sectional multilevel design of 201,221 adults nested within 50 US states we calibrated two-level binomial hierarchical mixed models (with states specified as a random effect). Our analyses suggest that for a 0.05 change in the state income inequality, the odds ratio (OR) of reporting poor health was 1.30 (95% CI: 1.17-1.45) in a conditional model that included individual age, sex, race, marital status, education, income, and health insurance coverage as well as state median income. With few exceptions, we did not find strong statistical support for differential effects of state income inequality across different population groups. For instance, the relationship between state income inequality and poor health was steeper for whites compared to blacks (OR=1.34; 95% CI: 1.20-1.48) and for individuals with incomes greater than $75,000 compared to less affluent individuals (OR=1.65; 95% CI: 1.26-2.15). Our findings, however, primarily suggests an overall (as opposed to differential) contextual effect of state income inequality on individual self-rated poor health. To the extent that contemporaneous state income inequality differentially affects population sub groups, our analyses suggest that the adverse impact of inequality is somewhat stronger for the relatively advantaged socioeconomic groups. This pattern was found to be consistent regardless of whether we consider contemporaneous or lagged effects of state income inequality on health. At the same time, the contemporaneous main effect of state income inequality remained statistically significant even when conditioned for past levels of income inequality and median income of states. PMID- 16338631 TI - Different public health geographies of the 2001 foot and mouth disease epidemic: 'citizen' versus 'professional' epidemiology. AB - Recently, there have been calls for health geographers to add critical and theoretical debate to 'post-medical' geographies, whilst at the same time informing 'new' public health strategies (Soc. Sci. Med. 50(9)1273; Area 33(4) (2002) 361). In this paper we reflect on how, alongside 'professional epidemiologies', 'citizen epidemiologies' can have credibility in informing public health policy and practice. We do this by drawing on mixed method and participatory research that used a citizens' panel to articulate the health and social outcomes of the 2001 foot and mouth disease disaster. We consider the difficulties of creating dialogue between on the one hand, time-limited, discrete, theoretical, visible and by implication legitimate, 'professional' knowledge and on the other, ongoing, holistic, experiential and often hidden 'citizen' knowledge of the foot and mouth disease epidemic. Despite significant evidence that in disaster and crisis situations, people need to be actively involved in key 'recovery' decisions (see for example At Risk Natural Hazards, People's Vulnerability, and Disasters, Routledge, London; A New Species of Trouble, Norton, New York), lay accounts, which may in themselves provide valuable evidence about the impact of the disaster, are often ignored. If health geographers are to critically inform 'new' public health policy then we need to consider research approaches that give voice to citizens' understanding of health outcomes as well as those of professionals. If 'new' public health is concerned with the material character of health inequalities, with fostering 'healthy' living and working environments, the promotion of community participation and individual empowerment (Area 33(4) (2002) 361), then we argue that situated, negotiated, everyday geographies of lay epidemiologies can and should inform public health policy. PMID- 16338632 TI - Urban asthma and the neighbourhood environment in New York City. AB - Asthma is now the leading cause of emergency room visits, hospitalizations, and missed school days in New York City's poorest neighbourhoods. While most research focuses on the influence of the indoor environment on asthma, this study examines the neighbourhood effects on childhood asthma, such as housing and ambient environmental hazards. Using Geographic Information Science (GI Science) we identify neighbourhoods with elevated concentrations of childhood asthma hospitalizations between 1997 and 2000 in US census tracts, analyze the sociodemographic, housing characteristics, and air pollution burdens from stationary, land use and mobile sources in these areas. The paper reveals the importance of distinguishing the specific and often different combinations of poor housing conditions, outdoor air pollution and noxious land uses that contribute to the high incidence of asthma in impoverished urban neighbourhoods. PMID- 16338633 TI - I'm not a doctor but I play one on TV: E.R. and the place of contemporary health care in fixing crisis. AB - This paper is an examination of the popular TV drama E.R. What is notable for health geographers about E.R. is how the show offers a representation of health care and the role of place in creating ways to provide care. Indeed, the place of health care-the emergency room-is the point of reference for the show's weekly dramas and centers the activity on the screen. We posit that the show's success stems from how crisis has become a central component of discourses about health care and that E.R. offers one highly seductive interpretation of how to deal with crisis in health care and care delivery. E.R. provides a representation of crisis by constructing three scales of intervention as the best sites to respond to and fix crisis: bodies, medical networks, and the urban social relations of the city. Order is designed around these scales which serve to map out where medical interventions can be made within the discursive regime of crisis. What E.R. provides is a powerful, if limited, "realistic" portrayal about the role of health care today-a role that is increasingly considered to be shaped by the need to intervene in crisis. PMID- 16338634 TI - Geographical variations in hospital use in Estonia. AB - The purpose of this study was to analyse the utilisation of inpatient care in Estonia. Using small-area analysis the age and sex of municipality residents and travel time to the nearest hospital were linked to the frequency of use and length of stay of respective inpatients. Adjusted hospitalisation and readmission rates decreased with increasing travel time to the hospital. The travel time effect on the length of stay was inconsistent. Despite differences in hospital utilisation among municipalities, geographical access to inpatient care was substantially diminished only for 3% of the population living further than 30 min from hospital. PMID- 16338635 TI - Environmental mercury release, special education rates, and autism disorder: an ecological study of Texas. AB - The association between environmentally released mercury, special education and autism rates in Texas was investigated using data from the Texas Education Department and the United States Environmental Protection Agency. A Poisson regression analysis adjusted for school district population size, economic and demographic factors was used. There was a significant increase in the rates of special education students and autism rates associated with increases in environmentally released mercury. On average, for each 1,000 lb of environmentally released mercury, there was a 43% increase in the rate of special education services and a 61% increase in the rate of autism. The association between environmentally released mercury and special education rates were fully mediated by increased autism rates. This ecological study suggests the need for further research regarding the association between environmentally released mercury and developmental disorders such as autism. These results have implications for policy planning and cost analysis. PMID- 16338636 TI - Rural/urban differences in accounts of patients' initial decisions to consult primary care. AB - In the UK, patients in urban areas consult primary health care more than rural patients for both trivial and serious conditions. This study, involving focus groups and interviews, examined rural/urban differences in accounts of patients' intentions around initial decisions to consult general practice. Findings suggest 'relationships' between doctors and patients and easier access to appointments could affect consulting in rural areas, while decision-making for urban patients tended to be more consumerist. Perceptions about access to different health services meant rural patients' decision-making in out-of-hours emergencies was complex. Rural/urban differences in demand could be affected by change in UK primary care provision. PMID- 16338637 TI - Restructuring home care in the 1990s: geographical differentiation in Ontario, Canada. AB - Through exploring the geographically differentiated effects of restructuring on the work of home care practitioners, labor process change is found to be spatially specific. It is through examining the spatiallity of labor process change that the differential impacts of health care restructuring on human health care resources can be determined. This has implications for human health care resource availability and service provision, which is particularly an issue for medically under-serviced regions. The restructuring approach, together with regulation theory, is used in this paper as tools for exploring the effects of health care reform on the local labor process. Both quantitative and qualitative data collected from home care practitioners in Ontario (Canada) show that the general trends in labor process change (work transfer down the home care hierarchy, increased time constraints, and enhanced job stress) are being similarly experienced, while shedding light on specific local differences. Findings show two working life factors to be differently experienced across the size of the place in which practitioners live and work. Two representative localities provide a closer examination of local differences in labor process change via the examination of local service cultures, local institutional practices, and local practitioner advocacy. The two sites are Sault Ste. Marie, located in northern Ontario's medically under-serviced northern health region, and Guelph, found in the province's well-serviced southern region. PMID- 16338639 TI - The assays of activities and function of TH, AADC, and GCH1 and their potential use in ex vivo gene therapy of PD. AB - In the past decades, there have been numerous studies in the gene therapy for Parkinson's disease (PD), especially in delivering genes of enzymes for dopamine (DA) synthesis. Gene therapy in PD appears to be at the brink of the clinical study phase. However, there are many questions that need to be solved before this approach can be contemplated clinically, especially the question about the control of DA production because too much DA could cause toxicity. Until recently, few studies have investigated the relation between DA production and PD improvement and respective expressed human tyrosine hydroxylase (hTH), human GTP cyclohydrolase 1 (hGCH1), and human aromatic acid decarboxylase (hAADC) in ex vivo gene therapy for PD. Now, we have developed a simple, fast, and reliable method to assay the activities of TH and AADC and have provided the possibility of ex vivo gene therapy for PD by genetically modifying cells with separate hTH, hGCH1, and hAADC genes. Using the method, we found though hTH, hGCH1, and hAADC genes were expressed, respectively, they could fulfil the function of DA synthesis by incubating together in vitro, and more DA was synthesized in vitro when hTH, hGCH1, and hAADC genes were expressed together rather than hTH and hAADC genes expressed or hTH expressed. The result suggests that we could easily control DA production in ex vivo gene therapy before transplantation. By combining this method and microdialysis, we also could further investigate the DA production in vitro and in vivo and then decide the optimal number and ratio of different transduced cells to improve the therapy of PD. Thus, the method has potential use in ex vivo gene therapy of PD. PMID- 16338640 TI - Learned helplessness: validity and reliability of depressive-like states in mice. AB - The learned helplessness paradigm is a depression model in which animals are exposed to unpredictable and uncontrollable stress, e.g. electroshocks, and subsequently develop coping deficits for aversive but escapable situations (J.B. Overmier, M.E. Seligman, Effects of inescapable shock upon subsequent escape and avoidance responding, J. Comp. Physiol. Psychol. 63 (1967) 28-33 ). It represents a model with good similarity to the symptoms of depression, construct, and predictive validity in rats. Despite an increased need to investigate emotional, in particular depression-like behaviors in transgenic mice, so far only a few studies have been published using the learned helplessness paradigm. One reason may be the fact that-in contrast to rats (B. Vollmayr, F.A. Henn, Learned helplessness in the rat: improvements in validity and reliability, Brain Res. Brain Res. Protoc. 8 (2001) 1-7)--there is no generally accepted learned helplessness protocol available for mice. This prompted us to develop a reliable helplessness procedure in C57BL/6N mice, to exclude possible artifacts, and to establish a protocol, which yields a consistent fraction of helpless mice following the shock exposure. Furthermore, we validated this protocol pharmacologically using the tricyclic antidepressant imipramine. Here, we present a mouse model with good face and predictive validity that can be used for transgenic, behavioral, and pharmacological studies. PMID- 16338645 TI - Endoscopic detection of early upper GI cancers. AB - The detection of early-stage neoplastic lesions in the upper GI tract is associated with improved survival and the potential for complete endoscopic resection that is minimally invasive and less morbid than surgery. Despite technological advances in standard white-light endoscopy, the ability of the endoscopist to reliably detect dysplastic and early cancerous changes in the upper GI tract remains limited. In conditions such as Barrett's oesophagus, practice guidelines recommend periodic endoscopic surveillance with multiple biopsies, a methodology that is hindered by random sampling error, inconsistent histopathological interpretation, and delay in diagnosis. Early detection may be enhanced by several promising diagnostic modalities such as chromoendoscopy, magnification endoscopy, and optical spectroscopic/imaging techniques, as these modalities offer the potential to identify in real-time lesions that are inconspicuous under conventional endoscopy. The combination of novel diagnostic techniques and local endoscopic therapies will provide the endoscopist with much needed tools that can considerably enhance the detection and management of early stage lesions in the upper GI tract. PMID- 16338646 TI - Pathology of early upper GI cancers. AB - Local endoscopic resection techniques for early neoplasms of the gastro intestinal tract require exact description of the depth of infiltration for the decision of endoscopic versus surgical therapy. Subdivision of mucosal neoplasms is used only in the oesophagus. Mucosal oesophageal carcinoma (squamous cell carcinoma and adenocarcinoma) can be subdivided into m1-m3 and m1-m4. Distinction of high-grade intraepithelial neoplasia and mucosal carcinoma is without clinical relevance since the diagnosis of high-grade intraepithelial neoplasia should always first lead to a (diagnostic) endoscopic resection. The final histological diagnosis could then be made on the resection specimen. Diagnosis of low-grade intraepithelial neoplasia is often confused with regenerative changes. Histological diagnoses of early neoplasms are not the same worldwide and consensus should be improved further. PMID- 16338647 TI - Endoscopic mucosal resection for early-stage gastrointestinal cancers. PMID- 16338648 TI - Endoscopic treatment of high-grade intraepithelial neoplasia and early cancer in Barrett oesophagus. AB - In the last 5 years, endoscopic therapy for high-grade intraepithelial neoplasia (HGIN) and early cancer (EC) in Barrett oesophagus has emerged as an effective and safe alternative to surgery. Adequate work-up of patients includes histopathological review of the initial biopsies, a high-resolution endoscopy with four-quadrant random biopsies every 1cm of Barrett mucosa and staging with endoscopic ultrasonography. Endoscopic resection (ER) forms the mainstay of the endoscopic treatment since it provides large tissue specimens for optimal histopathological evaluation. The ER-cap technique with submucosal injection and the 'suck-band-and cut' method are the resection methods most widely used in Barrett oesophagus patients. ER monotherapy for HGIN or EC in Barrett oesophagus is associated with recurrent lesions in up to 30% of treated patients. ER may be combined with ablative techniques such as photodynamic therapy (PDT) to treat all of the mucosa at risk for neoplastic progression. Unlike ER, PDT lacks histopathological correlation and residual Barrett mucosa may remain after treatment or may be hidden underneath the neosquamous epithelium. Management of Barrett oesophagus patients with HGIN or EC should be performed in centres with multi-disciplinary experience in this field and future studies should focus on development of ER techniques that allow radical resection of the whole Barrett segment. PMID- 16338649 TI - Endoscopic therapy of early gastric cancer. AB - Endoscopic therapy of early gastric cancer is applicable for differentiated-type mucosal carcinomas that have an extremely low potency of lymph-node metastasis. Among various kinds of endoscopic therapy, endoscopic mucosal resection is the most recommended procedure, because pathological evaluation of affected tissues is available using this method. Recently, endoscopic submucosal dissection, a novel method of endoscopic mucosal resection, has gained interest as a more reliable therapeutic procedure. In the present chapter several issues will be presented on endoscopic therapy for early gastric cancer, including endoscopic diagnosis of early gastric cancer, currently accepted indications of endoscopic therapy, and the possibility of extending the indication and techniques used for mucosal resection. PMID- 16338650 TI - Surgical strategies for early esophageal adenocarcinoma. AB - The need for extensive surgical resection for early-stage esophageal adenocarcinoma has been challenged by the increasing frequency of early detection in patients with Barrett's esophagus undergoing surveillance endoscopy. Limited endoscopic or surgical procedures are promoted as alternatives to radical esophagectomy and lymphadenectomy in such patients. Currently available data show that limited surgical resection of the distal esophagus with regional lymphadenectomy and interposition of an isoperistaltic jejunal segment is a safe and oncologically adequate procedure in this situation and provides good quality of life. This is in contrast to endoscopic ablation or endoscopic mucosal resection, which are associated with high tumour recurrence rates and persistence of premalignant Barrett esophagus. New technologies for accurate prediction of the presence and pattern of lymphatic spread-e.g. sentinel node techniques and artificial neural networks-may allow a further reduction of the invasiveness of surgical resection without compromising cure rates. PMID- 16338651 TI - Endoscopic detection of early lower gastrointestinal cancer. AB - The prognosis for patients with malignancies of the lower gastrointestinal tract is strictly dependent on early detection of premalignant and malignant lesions. What should an ideal screening and surveillance colonoscopy be able to accomplish? The technique should allow detection of large but also discrete mucosal alterations. Ideally, endoscopic discrimination between neoplastic and non-neoplastic lesions would be possible during the ongoing procedure. At present, endoscopy can be performed with powerful new endoscopes. Comparable to the rapid development in chip technology, the optical features of the newly designed endoscopes offer resolutions, which allow new surface details to be seen. In conjunction with chromoendoscopy, the newly discovered tool video colonoscopy is much easier and more impressive today than with the previously used fibre-optic endoscopes. Recently, new endoscopic technologies such as narrow band imaging, endocytoscopy, or confocal laser endoscopy have allowed the discovery of a whole new world of image details which will surely improve the diagnostic yield in the field of early malignancies. This review summarises newly available technologies and clinical data about the diagnosis of early lower gastrointestinal cancers. PMID- 16338652 TI - Pathology of early lower GI cancer. AB - Early colorectal cancer can be treated with curative resection if the depth of invasion is limited to the submucosa (pathologic T category pT1 in the TNM classification). Macroscopically early colorectal cancer and its precursor lesions present as elevated polyps or non-polypoid flat lesions. Microscopically, precursor lesions are characterized by intraepithelial neoplasia and present as classic adenomas or serrated adenomas. Precursor lesions may already contain foci of early colorectal cancer. Early colorectal cancer can be treated by endoscopic resection. Careful handling of the specimen is required in order to optimally identify the factors that may predict an adverse outcome. Whenever a favourable tumour grade is found, without vascular invasion and tumour budding, there seems to be a low risk for adverse outcome and laparotomy may thus be avoided. PMID- 16338653 TI - Endoscopic therapy of lower gastrointestinal cancer. AB - Since the 1960's, endoscopy has revolutionised the practice of gastroenterology. Although initially diagnostic, endoscopy is now playing an increasingly therapeutic role. There are many reasons to believe that therapeutic endoscopy will shape the practice of gastroenterology further in the future. Only a few years ago we relied on low-resolution fibreoptic endoscopes. Nowadays even standard equipment allows the mucosa to be scrutinised in great detail. Dedicated training in endoscopy together with attention to quality indicators such as polyp detection and caecal intubation rates will ensure that fewer early gastrointestinal cancers are missed in the future. Open access endoscopy and screening programs are being introduced in many Western countries which will also lead to more lesions being detected in their early stages. This chapter discusses the main issues surrounding the endoscopic therapy of lower gastrointestinal cancers. PMID- 16338654 TI - Minimally invasive surgery for early lower GI cancer. AB - Two technical developments in colorectal surgery-i.e. transanal endoscopic microsurgery (TEM) and laparoscopic surgery for colorectal disease-are now available for the treatment of early lower GI cancer. Benign lesions and early stage tumours of the rectosigmoid are amenable for a transanal approach. Transanal endoscopic microsurgery is performed using a rectoscope 4 cm in diameter with a four-port insert. After installation of a pneumorectum, lesions up to 25 cm from the anal verge, including circumferential lesions, can be removed with a recurrence rate of 0-5% for adenomas, 3% for low-risk T1 carcinomas, and 8% for all carcinomas. Laparoscopic-assisted colonoscopic polypectomy, laparoscopic wedge resection or laparoscopic-assisted colostomy have a 67-100% success rate for avoiding a formal bowel resection for benign tumours that cannot be treated by colonoscopy alone. Early colonic cancer requires laparoscopic colectomy guided by preoperative colonoscopy or preoperative endoscopic tattooing for localisation of the affected segment. PMID- 16338655 TI - Lasting longer without oxygen: The influence of hypoxia on Notch signaling. AB - Notch signaling has multiple functions during invertebrate and vertebrate development, where it plays important roles in cell fate determination, proliferation, survival, and border formation. The precise function of Notch signaling is context dependent, and one of the unsolved mysteries of Notch signaling is how a relatively stereotyped signal transduction pathway exerts such a wide variety of context-specific responses. Recent data from Gustaffson et al. provide important new information on this topic by showing that hypoxia enhances Notch signaling due to the association of Notch and HIF-1alpha. This interaction may have important consequences for tumor cell growth. PMID- 16338656 TI - A knotty turnabout?: Akt1 as a metastasis suppressor. AB - Akt is well known to enhance malignancy and is recognized as a key target for antineoplastic therapies. However, intriguing findings reported by Yoeli-Lerner et al. in the November 23, 2005 issue of Molecular Cell, suggest a novel, antimetastasis function of Akt: activation of Akt1 inhibited invasion in some cancer cells. One possible mechanism for this surprising phenotype was that Akt activated the E3 ubiquitin ligase HDM2, causing ubiquitination and degradation of NFAT, an invasion-promoting factor. These findings clearly justify further investigations and, if validated in vivo, call for reevaluation of some Akt targeting therapeutic strategies currently under development. PMID- 16338657 TI - Revealing the genomic heterogeneity of melanoma. AB - The melanoma genome possesses numerous recurrent chromosomal rearrangements, and embedded within this complexity are clues critical to disease pathogenesis and response to therapy. High-resolution genome-wide DNA copy number approaches, in conjunction with gene-specific mutational analyses, appear poised to define keystone molecular events, provide more accurate classification schemes, and set the stage for the design of rational therapies that may finally have an impact on survival of this deadly disease. PMID- 16338658 TI - The hypoxic microenvironment of the skin contributes to Akt-mediated melanocyte transformation. AB - Constitutive activation of Akt characterizes a high percentage of human melanomas and represents a poor prognostic factor of the disease. We show that Akt transforms melanocytes only in a hypoxic environment, which is found in normal skin. The synergy between Akt and hypoxia is HIF1alpha mediated. Inhibition of HIF1alpha decreases Akt transformation capacity in hypoxia and tumor growth in vivo, while overexpression of HIF1alpha allows anchorage-independent growth in normoxia and development of more aggressive tumors. Finally, we show that mTOR activity is necessary to maintain the transformed phenotype by sustaining HIF1alpha activity. Taken together, these findings demonstrate that Akt hyperactivation and HIF1alpha induction by normally occurring hypoxia in the skin significantly contribute to melanoma development. PMID- 16338659 TI - Mitogen requirement for cell cycle progression in the absence of pocket protein activity. AB - Primary mouse embryonic fibroblasts lacking expression of all three retinoblastoma protein family members (TKO MEFs) have lost the G1 restriction point. However, in the absence of mitogens these cells become highly sensitive to apoptosis. Here, we show that TKO MEFs that survive serum depletion pass G1 but completely arrest in G2. p21CIP1 and p27KIP1 inhibit Cyclin A-Cdk2 activity and sequester Cyclin B1-Cdk1 in inactive complexes in the nucleus. This response is alleviated by mitogen restimulation or inactivation of p53. Thus, our results disclose a cell cycle arrest mechanism in G2 that restricts the proliferative capacity of mitogen-deprived cells that have lost the G1 restriction point. The involvement of p53 provides a rationale for the synergism between loss of Rb and p53 in tumorigenesis. PMID- 16338660 TI - Kit-activating mutations cooperate with Spi-1/PU.1 overexpression to promote tumorigenic progression during erythroleukemia in mice. AB - The erythroleukemia developed by spi-1/PU.1 transgenic mice is a multistage process characterized by an early arrest of the proerythroblast differentiation followed later on by malignant transformation. Herein, we report the presence of acquired mutations in the SCF receptor gene (Kit) in 86% of tumors isolated during the late stage of the disease. Kit mutations affect codon 814 or 818. Ectopic expression of Kit mutants in nonmalignant proerythroblasts confers erythropoietin independence and tumorigenicity to cells. Using PP1, PP2, and imatinib mesylate, we show that Kit mutants are responsible for the autonomous expansion of malignant cells via Erk1/2 and PI3K/Akt activations. These findings represent a proof of principle for oncogenic cooperativity between one proliferative and one differentiation blocking event for the development of an overt leukemia. PMID- 16338661 TI - Decatenation checkpoint deficiency in stem and progenitor cells. AB - The decatenation checkpoint normally delays entry into mitosis until chromosomes have been disentangled through the action of topoisomerase II. We have found that the decatenation checkpoint is highly inefficient in mouse embryonic stem cells, mouse neural progenitor cells, and human CD34+ hematopoietic progenitor cells. Checkpoint efficiency increased when embryonic stem cells were induced to differentiate, which suggests that the deficiency is a feature of the undifferentiated state. Embryonic stem cells completed cell division in the presence of entangled chromosomes, which resulted in severe aneuploidy in the daughter cells. The decatenation checkpoint deficiency is likely to increase the rates of chromosome aberrations in progenitor cells, stem cells, and cancer stem cells. PMID- 16338662 TI - If I could change the world. PMID- 16338668 TI - Part 1: early recognition and treatment of birth trauma: injuries to the head and face. AB - The Joint Commission on Accreditation of Healthcare Organizations (JCAHO) Sentinel Event Alert from July 21, 2004 states that there have been 47 cases of birth trauma-related perinatal death or permanent disability reported for JCAHO review since 1996. This report clearly illustrates the importance of birth trauma in clinical practice for neonatal and perinatal nurses. Estimates suggest that birth trauma occurs in 2% to 7% of all deliveries and is associated with an increase in both mortality and morbidity. Birth trauma to the head may result in minor superficial extracranial injuries, such as caput succedaneum and cephalohematoma, or more serious and potentially life threatening lesions such as subgaleal hemorrhages. The potential for deeper intracranial injury, such as subarachnoid or subdural hemorrhage exists; these may be isolated or associated with skull fractures and/or other extracranial injuries. Injury to the eye, nasal structures, and paralysis of the vocal cords may also result from birth trauma during a difficulty delivery. Part 1 of this 2-part article will focus on birth injuries to the head and face. Part 2 of the series will review more systemic birth injuries that may involve abdominal organs, the spine and skeletal system, and peripheral and facial nerves. PMID- 16338669 TI - The clinical presentation of Ehlers-Danlos syndrome. AB - Ehlers-Danlos syndrome (EDS), a heterogeneous group of inheritable connective tissue disorders, is attributed to mutations in connective tissue genes. These mutations cause defects in collagen. Collagen, a connective tissue protein that acts like glue, gives strength to the body and provides support and elasticity for movement. Thus, the altered gene affects the mechanical properties of skin, joints, ligaments, and blood vessels. Ehlers-Danlos syndrome is transmitted through autosomal dominant, autosomal recessive, or x-linked patterns of inheritance. The life expectancy of an affected infant varies with the type of EDS. This article provides an overview of the 6 major classifications of EDS, their unique clinical presentations, a focused physical assessment guide, considerations for nursing care, and resources for parents. Ehlers-Danlos syndrome can be a potentially debilitating syndrome. It requires preventative and protective measures starting at birth to preserve joint function to improve infant outcomes. Caring for patients with EDS requires an understanding of the potential associated complications to help minimize the physical and emotional impact of the syndrome and improve the quality of life for affected individuals. PMID- 16338670 TI - Acculturation and stress in Chinese-American parents of infants cared for in the intensive care unit. AB - PURPOSE: To describe the stressful experiences of Chinese-American parents who have an infant in the intensive care unit (ICU), and to assess the influence of acculturation, parents' characteristics, and social supports on parental stress. DESIGN: A cross-sectional, descriptive, correlational study. SAMPLE/SETTING: A convenience sample of 30 Chinese-American families (30 mothers, 25 fathers) who had an infant hospitalized in 3 ICU sites in the San Francisco area. METHODOLOGY: Measures included: (1) Infant's health data from the medical records and parents' demographic data, and (2) Parental Stressor Scale: Infant Hospitalization, Suinn Lew Asian Self-Identity Acculturation Scale, and Family Support Scale. MAIN OUTCOME MEASURE: The dependent variable was mothers' and fathers' stress levels. Independent variables included personal/family characteristics, acculturation, uncertainty regarding infants' illness, and social supports. PRINCIPAL RESULTS: The ICU stressors were ranked by both mothers and fathers in the following order: infant's appearance, parental role alteration, staff communication and behaviors, and ICU environment. The combined effects of uncertainty about their infant's illness and its future impact, a strong belief in Asian family values, and lack of support from healthcare providers accounted for 26% and 55% of the variance in parental stress for mothers and fathers, respectively. CONCLUSIONS: Findings from this study suggest implications for the education of both families and the ICU staffs. PMID- 16338671 TI - Impacting infant head shapes. AB - Infant sleep position impacts the development of head shape. Changes in infant sleep position, specifically the movement toward supine sleep, have led to a redefinition of normal head shape for infants in the United States. Historically, a dolichocephalic (elongated) head shape was the norm. Currently the norm has changed to a more brachycephalic (shorter and broader) shape. Since the American Academy of Pediatrics' Back to Sleep Campaign, the incidence of positional plagiocephaly has increased dramatically with a concurrent rise in the incidence of torticollis. Infants who require newborn intensive care, particularly premature infants, are more prone to positional plagiocephaly and dolichocephaly. Both can be prevented or minimized by proper positioning. The infant with an abnormal head shape requires careful evaluation; treatment varies according to the etiology. Craniosynostosis, a less common but pathological etiology for plagiocephaly, should be considered in the diagnostic process. Successful treatment of positional plagiocephaly and dolichocephaly includes systematic positioning changes to overcome the mechanical forces of repetitive positioning, physical and/or occupational therapy to treat underlying muscle or developmental challenges, and in some cases, molding helmet therapy. PMID- 16338672 TI - Temporal lobe epilepsy in children: overview of clinical semiology. AB - Temporal lobe epilepsy in children has been less well studied compared to that seen in adults. Whereas hippocampal sclerosis is the most common etiology for the adult temporal lobe epilepsy syndromes, tumors and malformations of cortical development are more commonly seen in children. Differences in seizure semiology are also apparent. Temporal lobe epilepsy in very young infants may exhibit prominent motor manifestations reminiscent of extra-temporal seizures. These motor manifestations however decrease with increasing age and are less abundant in adults. Automatisms, which are commonly seen in temporal lobe epilepsy in all age groups, are simple at a younger age and become increasingly more complex and discrete with age. Several case studies, illustrated on the video are included in this review. They highlight the differences in temporal lobe seizure semiology between children and adults. [Published with video sequences]. PMID- 16338673 TI - A proposal for a five-dimensional patient-oriented epilepsy classification. AB - The recent proposal by the ILAE Task Force for Epilepsy Classification consists of a multi-axial syndrome-oriented approach. Epilepsy syndromes, as defined by the ILAE, group patients according to various, poorly defined parameters. The resulting syndromes have frequently no biological significance, with overlap among different syndromes and syndromes changing with age. Additionally, only a minority of patients can be classified syndromatically, and the axes of this classification system convey redundant information. We propose a five dimensional, patient-oriented approach to classifying epilepsies. This approach shifts from the syndrome-oriented approach to a standard, neurological, methodological, patient-oriented approach, using independent criteria in each of the five dimensions. Similar to general neurology, the first step in each patient physician encounter in epileptology is to take a history of the presenting symptoms and generate a hypothesis regarding the localization and etiology of the symptom within the nervous system. Therefore, the main dimensions of this classification consist of: 1) localization of the epileptogenic zone, 2) seizure semiology classified according to the semiological seizure classification, 3) etiology, 4) seizure frequency, and 5) related medical conditions. These dimensions characterize all of the information necessary for patient management, are independent parameters, and include more pertinent information with regards to patient management than the ILAE axes. All patients can be classified according to this five-dimensional system even at the initial patient encounter when no detailed test results are available. Information from clinical tests, such as MRI and EEG, are translated into the best possible working hypothesis at the time of classification, allowing for increasing precision of the classification as additional information becomes available. This patient-oriented classification envisions an epileptic seizure as an independent symptom of a central nervous system dysfunction due to different causes, with various cortical localizations, occurring at various frequencies, and in conjunction with other diseases and clinical symptoms. PMID- 16338674 TI - Developmental outcome of epilepsy surgery in tuberous sclerosis complex. AB - In Tuberous sclerosis complex (TSC), neurological dysfunction, usually in association with epilepsy, is responsible for the greatest degree of disease related disability. Epilepsy surgery is increasingly recognized as a therapeutic option given the often medication-resistant nature of the disease. Seven subjects with medically refractory epilepsy associated with TSC, who underwent surgery at a tertiary care epilepsy center and in whom both preoperative and postoperative neuropsychological data were available, were examined. The Vineland Adaptive Behavior Scales, and in one case, the WISC-III were utilized. Postoperatively, the composite standard scores declined in six of the seven subjects, although for the most part this decline was quite modest (8 points or less in 5/6 subjects). The mean overall developmental/intellectual quotients were comparable across assessments (preoperative M = 55, SD = 20.3; postoperative M = 49 SD = 16.6). Good outcomes appeared to be related to seizure relief. Age estimates of developmental level indicated developmental progress in the majority of subjects in the current sample, and may yield greater clinical information for individuals with developmental delay than do standard scores. PMID- 16338675 TI - Tonic status epilepticus in patients with idiopathic generalized epilepsy. AB - RATIONALE: Tonic status epilepticus (TSE) in patients with idiopathic generalized epilepsy (IGE) is not well recognized. The objective of this study is to report episodes of TSE in patients with IGE. METHODS: We retrospectively reviewed the clinical and EEG evaluation of three IGE patients who presented TSE. RESULTS: The three patients had mainly clinical features of IGE, but had developed, in addition, focal discharges, diffuse EEG abnormalities and some focal or diffuse neuropsychological dysfunction. The tonic attacks eventually responded to treatment, but were not completely controlled in any of the patients. DISCUSSION: The continuum between IGE and secondary generalized epilepsy is demonstrated in these patients. Most of their clinical and EEG features are however, in keeping with an idiopathic generalized epileptic process with additional focal and diffuse components. Recognition of the significance of TSE in such patients has important therapeutic and prognostic implications. [Published with video sequences]. PMID- 16338676 TI - Ictal video-polysomnography and EEG spectral analysis in a child with severe Panayiotopoulos syndrome. AB - OBJECTIVE: To describe the ictal polysomnographic features of a patient with Panayiotopoulos syndrome, a peculiar epileptic syndrome characterized by infrequent, often single, prolonged, nocturnal, focal seizures comprising an unusual constellation of autonomic symptoms (malaise, nausea, pallor, tachycardia, vomiting) and unilateral deviation of the eyes at the onset of seizures. These clinical, ictal manifestations are rarely followed by post-ictal headache. In the literature, there is little information on the ictal EEG characteristics of Panayiotopoulos syndrome and, in particular, on certain autonomic manifestations, such as tachycardia, as the sole ictal phenomena at the onset of seizures. METHODS AND RESULTS: One, all-night videopolysomnography, during which one seizure was recorded. Video-EEG data were evaluated visually and by means of quantitative spectral analysis. The spectral analysis of the recorded seizure showed a complex ictal pattern of cortical involvement with focal onset in the right occipital area followed by the recruitment of widespread extra occipital cortical regions. CONCLUSIONS: This is the first such analysis of this peculiar epileptic condition. Most of the symptoms were consistent with a diagnosis of severe Panayiotopoulos syndrome, although the patient also presented "atypical findings": a relatively high frequency of seizures, post-ictal headache, no spontaneous remission of seizures with age, and late onset of visual hallucinations; this last finding is more frequent in "Gastaut-type childhood occipital epilepsy", in which onset typically occurs later than in Panayiotopoulos syndrome. [Published with video sequences]. PMID- 16338677 TI - Late-onset, "Gastaut type", childhood occipital epilepsy: an unusual evolution. AB - We report on two girls and one boy with clinical and electroencephalographic features of late-onset childhood epilepsy with occipital paroxysms of the "Gastaut type", showing an unusual evolution. Neurological examination and brain imaging were normal in all three. At the age of 7.5 years, eight years and ten years respectively, the three children presented with episodes of visual symptoms when awake, and in one of them, the seizures were occasionally followed by oculocephalic deviation. The interictal EEG showed bilateral occipital spike-wave activated by eye closing. In two patients, the occipital seizures had been immediately followed by typical absences, since onset; in the other patient, five months after onset. The ictal EEG showed irregular bilateral occipital spike-wave discharges during the visual symptoms, followed by generalized spike-wave activity during the typical absences. The typical absences were activated by hyperventilation; the EEG did not show continuous spikes and waves during slow sleep. These three patients, with typical electroclinical features of "Gastaut type", childhood occipital epilepsy, demonstrated an evolution which, to our knowledge, has not been previously described. We investigated whether this unusual, age-dependent evolution was due to secondary bilateral synchrony or if these electroclinical features represent two types of idiopathic epileptic syndromes in the same patients. PMID- 16338678 TI - Subjective signs in premotor epilepsy: confirmation by stereo electroencephalography. AB - Subjective manifestations inaugurating frontal seizures are less well known than those observed in temporal seizures. We report eleven consecutive patients who underwent surgery for premotor epilepsy. Six of them had focal cortical dysplasia. Ictal symptomatology was analysed to establish electroclinical correlations. The localisation of the epileptogenic zone was assessed by stereoelectroencephalographic studies. Subjective manifestations were described in all cases, more frequently in a sensory rather than an emotional or psychological fashion. Focal seizures limited to subjective features were recorded in two patients. In one, psychological illusions and visual hallucinations were related to the superior frontal sulcus. Another presented isolated paraesthesia in the left arm with the implication of the supplementary motor area. Electrical stimulation of an electrode located in the premotor area evoked isolated subjective manifestations in three other patients. One patient reported sensory manifestations and another, ideational manifestations. Cephalic sensations and emotional manifestations were associated in one case. Subjective manifestations were observed in all patients, and were proved to be related to a discharge restricted to the premotor area in five. These were non-specific signs, but were always the same in a given patient. Spontaneous, isolated sensations and stimulation data tended to be contradictory. This illustrates the complexity of analyzing subjective signs, as well as the complexity of the neuronal networks participating in the propagation of discharges arising in the premotor frontal area. PMID- 16338679 TI - Spike frequency is dependent on epilepsy duration and seizure frequency in temporal lobe epilepsy. AB - OBJECTIVES: We wanted to investigate factors that are associated with frequency of interictal epileptiform discharges by investigating 303 patients with temporal lobe epilepsy (TLE). METHODS: We included all patients who consecutively underwent the adult presurgical evaluation program at our center and who had intractable, medial TLE with complex partial seizures due to unilateral medial temporal lobe lesions. The interictal EEG samples were automatically recorded and stored on computer. The location and frequency of interictal epileptiform discharges were assessed by visual analysis of interictal EEG samples of 2-minute duration every hour. RESULTS: There were 303 patients (aged 16-63) who met the inclusion criteria. The median interictal epileptiform discharge frequency was 15 IED/h, the median seizure frequency was 4 seizures/month. According to univariate analyses, we found that age at monitoring, epilepsy duration, and higher seizure frequency were associated with higher interictal epileptiform discharge frequency. In the logistic regression analysis, we found that higher seizure frequency (p < 0.001) and longer epilepsy duration (p = 0.007) were independently associated with higher spike frequency, while the age at monitoring was not. CONCLUSIONS: Seizure frequency and epilepsy duration (years of patient's life with seizure activity) were independently associated with IED frequency, suggesting that IED are modulated by seizures. PMID- 16338680 TI - Familial rectal pain: a familial autonomic disorder as a cause of paroxysmal attacks in the newborn baby. AB - A 2-day-old baby exhibited impressive paroxysmal attacks consisting of bradycardia, bronchospasm and vasomotor fits (Harlequin type) related to a rare, dominantly inherited form of dysautonomy called "familial rectal pain". These events were recurrently triggered by emotion, diaper changes or wiping of the perineal areas or eating. Sometimes they occurred spontaneously. Carbamazepine had an excellent effect on the fainting. At four years of age, the child had normal psychomotor development with only minimal symptoms, and very rare paroxysmal attacks. The father of the child has minimal symptoms of this entity, with essentially ocular manifestations. Familial rectal pain is a very rare entity that must always be considered as a possible aetiology of any life threatening event in an infant because of the availability of a very effective treatment. The existence of minimal forms of familial rectal pain is possible, and it is likely that this entity is underdiagnosed. [Published with video sequences]. PMID- 16338681 TI - A comparative study of mismatch negativity (MMN) in epilepsy and non-epileptic seizures. AB - This study investigated mismatch negativity (MMN) differences between subjects with non-epileptic seizures (NES), subjects with epilepsy, and healthy controls. Event-related potentials (ERPs) were obtained from 14 patients with NES, 15 patients with epilepsy and 16 healthy control subjects. A conventional MMN procedure was used with a random sequence of 12% deviant tones (922 Hz) and 88% standard tones (1000 Hz). Subjects were instructed to ignore the tones delivered through headphones whilst reading a book. Significant differences in distribution of the mismatch negativity (MMN) in patients with NES compared to controls were obtained (F3, p 0.05). In summary, the lack of agreement in profiles for temperature and the accuracies of long and short service suggests that some factor(s) that deteriorate during the course of the time spent awake might be involved, in addition to body temperature, in determining accuracy in the tasks measured. PMID- 16338716 TI - Influence of time of day and partial sleep loss on muscle strength in eumenorrheic females. AB - Disrupted sleep is the most common form of sleep deprivation in travellers, shift workers, athletes the night before important competitions and among parents of infants. The influence of partial sleep loss on muscle strength might differ according to the time of testing on the following day. This study was therefore designed to assess the interaction between the effects of partial sleep loss and time of day on muscle strength in females. Eight sedentary eumenorrheic females (mean +/- SD; age 30 +/- 6 years, height 1.62 +/- 0.06 m and body mass 67 +/- 5.0 kg) took part in the study, in a counterbalanced design. Measurements of muscle strength were carried out at 06:00 and 18:00 hours after the one control night (no sleep loss) and the one night of partial sleep loss, during menses. Muscle strength measures included isokinetic (at 1.05, 3.14 rad s(-1); 90 degrees range of motion) and isometric peak torque (at 60 degrees of knee flexion) of knee extensors and flexors (dominant leg). In addition, isometric force of knee extensors with super-imposed electrical twitches (50 Hz, 250 V, 200 mus pulse width) was measured using the same procedure in order to control for motivation. Rectal temperature was measured during the 30 min before muscle strength measurements. Partial sleep loss consisted of allowing 2.5 h sleep (between 03:00 and 05:30 h), whilst in the control condition (no sleep loss) subjects retired between 22:30 and 23:30 h, rising at 05:30 hours. All measurements were conducted at just one phase of the menstrual cycle (menses) to prevent any masking effect due to different phases of the menstrual cycle. In both conditions (with and without partial sleep loss) a diurnal variation was observed in peak torque of knee flexors at 1.05 (F(1,7) = 5.5, p < 0.05) and 3.14 rad s(-1) (F(1,7) = 8.0, p < 0.05); values at 18:00 hours were 4.5 and 5.9% higher than at 06:00 hours, respectively. No significant diurnal variation was observed for the other muscle strength measures. No significant effect of partial sleep loss or interaction effect (sleep x time of day) was observed for muscle strength measures. However, the performance rhythms were in phase with the circadian rhythm in rectal temperature. Partial sleep deprivation over one night did not have any adverse effect on maximal muscle strength, nor on diurnal variations of muscle strength indices. As the effect of time of day was observed with some of the muscle strength measures, it is suggested that, in designing future studies using females, the control of time of day is essential. PMID- 16338717 TI - Effects of daytime ingestion of melatonin on short-term athletic performance. AB - The immediate effects of ingesting melatonin in the daytime include decreased alertness and body temperature. To date, no researcher has examined whether daytime ingestion of melatonin leads to impairments in variables relevant to short-term (<10 min) athletic performance. Twelve physically active participants (mean +/- s age = 25.2 +/- 5.0 years, body mass = 81.4 +/- 12.1 kg and chronotype = 33.8 +/- 6.3 units) ingested 5 mg of melatonin or placebo at 11:45 hours in a double-blind experiment. At 13:00 and 17:00 hours, subjective alertness was measured, together with intra-aural temperature, reaction time (two-, four- and eight-choice), short-term memory recall and grip strength. Performance, ratings of perceived exertion (RPE) and heart rate were also recorded during a 4-km cycling time trial. At 13:00 hours, the mean +/- s intra-aural temperature was 0.49 +/- 0.79 degrees C lower after ingestion of melatonin than after placebo (p = 0.015), but this difference was not apparent at 17:00 hours. At both 13:00 and 17:00 hours, melatonin reduced (p < 0.05) alertness, short-term memory and exercise heart rate by 1.5 +/- 1.8 units, 1 +/- 1 digits and 6 +/- 9 beats.min( 1), respectively (mean +/- s). Eight-choice reaction time was also slower at both times of day after ingesting melatonin. Melatonin did not influence time trial performance or RPE (p > 0.05). The effects of 5 mg of melatonin seem more pronounced for mental rather than physical components of short-term athletic performance, although the cardiovascular responses to exercise are affected. Some effects of melatonin were apparent 5 h after ingestion when the hypothermic effects of melatonin had dissipated. PMID- 16338718 TI - Effect of five nights of normobaric hypoxia on cardiovascular responses to acute isocapnic hypoxia in humans: relationship to ventilatory chemosensitivity. AB - The aim of this study was to elucidate (1) the cardiovascular responses to acute isocapnic hypoxia stimuli following five nights of normobaric poikilocapnic hypoxia, and (2) whether the changes in the cardiovascular responses to hypoxia are correlated to the change in acute hypoxic ventilatory (AHVR) chemosensitivity. Twelve male subjects [26.6 +/- 4.1 (SD) years] slept 8-9 h day( 1) overnight for five consecutive days at a simulated altitude of 4300 m (FiO2 = approximately 13.8%). Using the technique of dynamic end-tidal forcing, the AHVR was assessed twice, prior to and immediately after the hypoxic exposure. During each AHVR test, mean arterial blood pressure (MAP) and heart rate (HR) were measured continuously using finger photoplethysmography and an ECG monitor, respectively. Immediately following the exposure, AHVR and MAP sensitivities were increased by 1.80 +/- 1.30 l min(-1) %(-1) (p < 0.01) and 0.69 +/- 0.40 mmHg %( 1) (p < 0.05), respectively, and there were significant correlations between the increases in the AHVR and MAP sensitivities (r = 0.67; p < 0.05). In summary, following five nights of normobaric hypoxia, there is an enhanced MAP response to hypoxic stimuli. The relationship between the enhanced AHVR and MAP sensitivity raises the possibility of a common pathway in the regulation of peripheral chemosensitivity and MAP responses during periods of isocapnic hypoxia. PMID- 16338719 TI - Effects of intermittent hypoxic training on aerobic and anaerobic performance. AB - The aim of the present study was to determine whether short-term intermittent hypoxic training would enhance sea level aerobic and anaerobic performance over and above that occurring with equivalent sea level training. Over a 4-week period, two groups of eight moderately trained team sports players performed 30 min of cycling exercise three times per week. One group trained in normobaric hypoxia at a simulated altitude of 2750 m (F(I)O2= 0.15), the other group trained in a laboratory under sea level conditions. Each training session consisted of ten 1-min bouts at 80% maximum workload maintained for 2 min (Wmax) during the incremental exercise test at sea level separated by 2-min active recovery at 50% Wmax. Training intensities were increased by 5% after six training sessions and by a further 5% (of original Wmax) after nine sessions. Pre-training assessments of VO(2max), power output at onset of 4 mM blood lactate accumulation (OBLA), Wmax and Wingate anaerobic performance were performed on a cycle ergometer at sea level and repeated 4-7 d following the training intervention. Following training there were significant increases (p < 0.01) in VO(2max) (7.2 vs. 8.0%), Wmax (15.5 vs. 17.8%), OBLA (11.1 vs. 11.9%), mean power (8.0 vs. 6.5%) and peak power (2.9 vs. 9.3%) in both the hypoxic and normoxic groups respectively. There were no significant differences between the increases in any of the above-mentioned performance parameters in either training environment (p > 0.05). In addition, neither haemoglobin concentration nor haematocrit were significantly changed in either group (p > 0.05). It is concluded that acute exposure of moderately trained subjects to normobaric hypoxia during a short-term training programme consisting of moderate- to high-intensity intermittent exercise has no enhanced effect on the degree of improvement in either aerobic or anaerobic performance. These data suggest that if there are any advantages to training in hypoxia for sea level performance, they would not arise from the short-term protocol employed in the present study. PMID- 16338720 TI - Exercise thermoregulation and hyperprolactinaemia. AB - The anterior pituitary hormone prolactin (PRL), measured in the peripheral blood circulation, reflects alterations in central brain 5-hydroxytryptamine (serotonin) and dopaminergic activity and is used as a marker of 'central fatigue' during active heat exposure. Significant correlations have consistently been found between PRL and core temperature (T(CORE)) during prolonged exercise. There has been no investigation into the relationship between PRL and other key thermoregulatory variables during exercise, such as weighted mean skin (T(SK)) and mean body temperature (T(B)), heat storage (HS), thermal gradient (T(GRAD)), heart rate (HR) and skin blood flow (cutaneous vascular conductance, CVC). Therefore, the aim of this study was to ascertain if a significant relationship exists between PRL and these thermoregulatory variables during prolonged exercise. Nine active male subjects conducted three trials of approximately 60% VO(2peak) at 70-80 rpm for 45 min on a semi-recumbent cycle ergometer at three different ambient temperatures [6 degrees C (Cold), 18 degrees C (Neutral) and 30 degrees C (Hot)] to elicit varying levels of thermoregulatory stress during exercise. Significant differences existed in T(SK), T(B), HS, T(GRAD) and CVC across the environmental conditions (p < 0.001). Core temperature (T(CORE)), HR and PRL were significantly elevated only in Hot (p < 0.05). Moderate correlations were found for T(CORE), T(SK), T(B), HS, T(GRAD), HR and CVC with post-exercise PRL (rho = 0.358-0.749). The end-of-exercise <38.0 degrees C T(CORE) responses were not (rho = -0.129, p > 0.05) but the >38.0 degrees C T(CORE) responses were (rho = 0.845, p < 0.001) significantly related to their corresponding PRL responses. The significant relationships between PRL release and T(SK), T(B), HS, T(GRAD), HR and CVC have extended previous research on T(CORE) and PRL release and indicate an association between these thermoregulatory variables, as well as T(CORE), and serotonergic/dopaminergic activity during prolonged exercise. PMID- 16338721 TI - Influence of environmental temperature on duathlon performance. AB - The aim of this study was to evaluate the physiological, metabolic and performance responses to duathlon performance under a range of ambient temperatures. Ten male recreational athletes performed three self-paced duathlon time trials consisting of a 5 km run (R1), a 30 km cycle and a 5 km run (R2) at 10 degrees C, 20 degrees C and 30 degrees C and a relative humidity of 50%. Performance times, heart rate (HR), rating of perceived exertion (RPE), core temperature (Tc) and skin temperature (Tsk) were measured every kilometre. Carbohydrate and fat oxidation rates were calculated via expired gas analysis at the first and fourth kilometres during both running stages. Blood samples were taken before and after exercise for the determination of prolactin concentration.Overall performance was significantly faster at 10 degrees C (100.76+/-5.32 min) than at 30 degrees C (105.38 +/- 4.28 min). Significantly higher Tc was noted in the 30 degrees C trial than in the 10 degrees C trial, with concomitant elevations in prolactin after exercise (19.88 +/- 6.48 ng/ml at 30 degrees C; 13.10 +/- 8.75 ng/ml at 10 degrees C). The rates of carbohydrate oxidation did not differ between conditions, although fat oxidation rates were highest at 10 degrees C. Elevated ambient temperature has a negative effect on duathlon performance. This effect may be reflected in increased Tc and prolactin concentration. PMID- 16338722 TI - Comparison of muscle strength and flexibility between the preferred and non preferred leg in English soccer players. AB - Most soccer players have a favoured foot for kicking the ball, and it is believed that this preference may lead to an asymmetry in the strength and flexibility of the lower extremities. This study was designed to determine whether asymmetry in strength and flexibility are present in the legs of soccer players.Forty-one elite and sub-elite soccer players (age 23.4 +/- 3.8 years; height 1.81 +/- 0.06 m; body mass 81.7 +/- 9.9 kg) were studied (data are presented as mean +/- SD). The dynamic strength of knee flexors (hamstrings) and knee extensors (quadriceps) was measured using an isokinetic dynamometer at angular velocities of 1.05, 2.09, 5.23 rad/s (in a concentric mode) and 2.09 rad/s (in an eccentric mode). The concentric strength ratio (hamstrings(conc)/quadriceps(conc)) and the dynamic control ratio (hamstrings(ecc)/quadriceps(conc)) were computed. Hip joint flexibility (in flexion) was measured using a goniometer.A significant difference between the preferred and non-preferred leg was found in the knee flexors at 2.09 rad/s (119 +/- 22 versus 126 +/- 24 Nm; P < 0.05) and for the dynamic control ratio (0.79 +/- 0.13 versus 0.84 +/- 0.16 Nm; P < 0.05). In both cases the knee flexors of the preferred leg were weaker than those of the non-preferred leg. A total of 28 of the 41 players (68%) had significant musculoskeletal abnormality (imbalance >10%) in one or more specific muscle groups. No significant differences were found in flexibility of the hip joint between the preferred and non-preferred leg (P > 0.05). It is concluded that the lower strength of the knee flexor muscles of the preferred leg may be associated with the differential use of these muscle during the kicking action and thus constitutes a unique training effect associated with soccer. This in turn can lead to muscular imbalance which is generally regarded as an injury risk factor. PMID- 16338723 TI - Changes in the force development characteristics of muscle following repeated maximum force and power exercise. AB - There is some evidence to suggest that ability to develop force in skeletal muscle can be improved for a short period following repeated maximal strength exercise. Some studies have suggested the phenomenon known as post-activation potentiation (PAP) to explain this immediate response to such exercise. However, little is known about the physical and temporal characteristics of the changes in muscle function due to PAP when induced using whole-body maximal strength exercise. The purpose of this study was to establish the physical and temporal characteristics of PAP of muscle function in elite male athletes following maximum strength and power exercise. Fifteen participants performed one of repeated maximal strength exercise or maximal power exercise or a control protocol on separate occasions. Changes in maximum isometric voluntary force (iMF), maximum isometric rate of force development (iRFD) and maximum vertical counter-movement jump (CMJ) were assessed. Results suggest that post-activation potentiation of iRFD occurs but is preceded by an initial suppression of force development capability following repeated maximal strength exercise. Following repeated maximal power exercise, however, post-activation potentiation of iRFD occurs immediately but is not seen over as long a period as that observed following maximal strength exercise. These results suggest that post-activation potentiation does bring about increased force development capability following intense exercise and that the temporal profile of the post-activation potentiation changes with intensity of the exercise used to induce it. PMID- 16338724 TI - Effect of weightlifting upon left ventricular function and markers of cardiomyocyte damage. AB - The purpose of this study was to assess left ventricular (LV) function and biochemical markers of myocyte after prolonged weightlifting activity. Seventeen male subjects (age range 20-34 years) performed a 90-min bout of weightlifting exercise consisting of three sets of 8-10 repetitions at 70% one-repetition maximum. Body mass, heart rate, systolic blood pressure (SBP) and echocardiographically determined indices of LV loading (LV internal diameter during diastole, LV meridonial wall stress), systolic function (stroke volume (SV), ejection fraction (EF), end-systolic pressure volume relationship; SBP/ESV) and diastolic filling (ratio of early to late; E:A) were obtained pre-exercise, immediately after and 24 h post-exercise. A 5-ml venous blood sample was obtained for the assessment of cardiac troponin T (cTnT) via third generation electrochemiluminescence assay. Data were assessed via one-way ANOVA and Pearson's correlation. Although SV declined (80.9 +/- 18.3 vs. 66.9 +/- 17.2, p < 0.05) there was no alteration in LV contractility (EF 62 +/- 6 vs. 59 +/- 7; SBP/ESV 3.51 +/- 1.4 vs. 3.51 +/- 1.4, p > 0.05). The E:A ratio was significantly decreased following exercise (1.78 +/- 0.41 vs. 1.33 +/- 0.37, p < 0.05). This decrease was not fully explained by loading conditions (r2 = 0.05 to 0.24). All values returned to baseline 24 h post-exercise. No cTnT was reported in any of the blood samples. In conclusion, there was no significant evidence of any LV contractile depression and no cTnT was observed post exercise. The small reduction in diastolic filling could not be explained by changes in haemodynamic loading or the post-exercise elevation in heart rate. PMID- 16338725 TI - A biomechanical analysis of good and poor performers of the vertical jump. AB - The vertical jump is widely used as a field test of performance capability, particularly in games like soccer. Invariably some players perform better than others and, while this is usually put down to greater strength or 'explosive power', there is no detailed information to explain how the muscles around the major joints contribute to this performance and what the nature of this contribution is, or indeed whether aspects of technique are important to performance. Detailed knowledge of this type would be useful to help understand which muscle characteristics are important in successful performance of jumping and may enable insights to be gained in terms of strength training for players. The aim of this study was to investigate the contribution made by the lower limb joints to vertical jump performance by good and poor performers of the counter movement jump. Two groups of players were selected who were found to be good and poor jumpers, respectively. Each player was required to perform three maximal vertical counter-movement jumps with, and three jumps without, an arm swing. The jump performance was recorded simultaneously by means of a force platform and a ProReflex automatic motion analysis system at 240 Hz. Values at the ankle, knee and hip were computed from these data for joint moments and power.Generally, better jumpers demonstrated greater joint moments, power and work done at the ankle, knee and hip, and as a result jumped higher under both conditions. It appears that the superior performance of the better jumpers was due to greater muscle capability in terms of strength and rate of strength development in all lower limb joints rather than to technique, which differed less noticeably between the groups. It is concluded that the muscle strength characteristics of the lower limb joints are the main determinant of vertical jump performance with technique playing a smaller role. PMID- 16338726 TI - Electromyographic investigation of abdominal exercises and the effects of fatigue. AB - Abdominal exercises are widely used to develop the anterior muscles of the trunk. These exercises can be undertaken without the aid of equipment, but increasingly manufacturers are developing equipment which purportedly enhances the training effect for abdominal muscles. As there are many different products and exercises used for abdominal muscle development, it is likely that some are more effective than others. This study aimed to investigate the effectiveness of five commonly performed abdominal exercises. A second aim was to investigate the effects of fatigue on these exercises. Five different types of abdominal exercise [standard crunch (sit-up) with bent knees, gym ball crunch, crunch with 5 kg weight held behind the head, legs raised crunch and a commercially manufactured roller crunch] were examined using integrated surface electromyography (IEMG). The lower rectus abdominis (LRA), upper rectus abdominis (URA) and obliquus externus abdominis (EO) of 15 healthy male participants [age (mean +/- SD) 22.2 +/- 6.8 years; height 1.77 +/- 0.06 m; mass 79.3 +/- 10.7 kg] were monitored using a four channel special purpose EMG data logger. Three trials of each exercise were performed in random order and normalized to enable comparisons between muscles and exercises. At a later date, ten participants were then re-tested when fresh and after a 30 min whole-body fatigue protocol that specifically targeted the abdominal muscles. Two exercises were evaluated, the abdominal roller crunch and legs raised crunch, which were judged to be the least and most effective, respectively, of the five exercises previously used. The normalized IEMG showed significant (p < 0.001) differences between exercises (gym ball crunch = 86.0 +/- 7.5%; legs raised crunch = 79.9 +/- 5.1%; 5 kg weight crunch = 65.1 +/- 13.4%; standard crunch = 56.2 +/- 3.2%; and roller crunch = 45.0 +/- 11.4%). Post fatigue, the normalized mean IEMG for both exercises increased significantly (p < 0.05) for LRA and URA muscles but not for the EO (p > 0.05). It was concluded that exercises can be constructed to provide a greater challenge to abdominal muscles, commercially available roller-type equipment appears to be little different from the standard abdominal crunch with bent knees, and fatiguing exercise results in the LRA and URA being more highly activated. These findings provide more detailed knowledge and understanding of the effects of different forms of abdominal exercise. PMID- 16338727 TI - Repeatability of stature measurements in individuals with and without chronic low back pain. AB - Measurements of reduction in stature have been used to compare spinal loading in chronic low-back pain (CLBP) and asymptomatic populations. Whether there are any differences in the repeatability of stature measurements, between those with and without CLBP, is not known. This investigation aimed to determine the repeatability of stature measurements in those with (n = 12) and without (n = 12) CLBP, and to establish if the ability to produce repeatable measurements is retained after a specific timeframe. Stature measurements were taken on two separate sessions that were 2 weeks apart, using a stadiometer accurate to 0.01 mm. All participants attained a mean SD of < or = 0.5 mm by the third measurement set taken on the first session of testing and no significant difference in mean SD was found between those with (0.37 mm) and without (0.40 mm) CLBP (p > 0.05). Intraclass correlation coefficients (ICC) demonstrated good levels of repeatability for all stature measurements obtained from the participants and the values for Standard error of the measurement (SEM) improved as the mean SD decreased with each measurement set. Investigators should have confidence in the ability of those with and without CLBP to produce equally repeatable stature measurements with appropriate prior practice. The second session of testing demonstrated that both groups had retained the ability to achieve the desired level of repeatability (SD < or = 0.5 mm) 2 weeks later without further practice. PMID- 16338728 TI - High-speed non-invasive measurement of tibial rotation during the impact phase of running. AB - Several injuries to the lower extremity in runners have been linked to excessive rates of internal rotation of the tibia just after ground impact. This study presents an improved method of capturing internal/external tibial rotation, and investigates whether estimates of rates of internal tibial rotation during the first 50 ms of contact during running are influenced by the sampling rate and processing of tibial displacement data. A lightweight plate was moulded to the shape of each subject's right anterior-medial tibia. Nine male subjects ran barefoot (3.35 m.s(-1)) and the landing kinematics of the right leg were recorded at 1000 Hz. The group mean value for the total range of internal rotation of the tibia for the whole stance phase was consistent with the literature (15.4 degrees ), but peak angular velocities (8.3 rad.s(-1)) were substantially higher than previously reported. The cut-off frequency of the low-pass filter influenced the peak angular velocity values obtained with the largest changes occurring between 15 and 40 Hz. Typically, researchers using lower sample rates have to filter around 10 Hz and consequently are likely to underestimate peak angular velocities. These findings have implications for obtaining a sound quantitative foundation for transient tibial motion before furthering our understanding of injury mechanisms. PMID- 16338729 TI - Measurement of three-dimensional intra-articular kinematics: methodological and interpretation problems. AB - Intra-articular kinematics evaluates joint motion in terms of the configuration of the joint. Therefore data are needed concerning joint kinematics as well as joint configuration. We have developed accurate measurement methods for both in vivo and in vitro evaluation. Interpretation of the processed data is more complex than simply setting up a coordinate system based on the joint configuration. Although the description of intra-articular motion in terms of Euler-Cardan or helical angles may be complete, the therapeutic interpretation may be doubtful. Using the the ulno-humeral joint during flexion-extension as an example, we found the combination of helical angles in the directions of extension/external rotation/varus. In the case of the Cardan angles, inconsistent patterns of rotation resulted from a different choice of sequence order and were different from the helical angles. The finite helical axis (FHA) provides a functional representation of the joint movement, i.e. pathways of motion, whereas the sequence dependency of Euler-Cardan angles produces problems in the therapeutic interpretation of the movement. Therefore we believe that an FHA approach should be used in intra-articular kinematics research. PMID- 16338730 TI - Arthro-kinematics of the elbow: study of the carrying angle. AB - The carrying angle of the elbow is usually assessed in full elbow extension, with a protractor goniometer, or derived from X-ray images. Substantial differences in carrying angle values have been reported, possibly explained by methodological differences. Carrying angles tend to show higher values in women than in men. The aim of this study was to confirm the previously described progressive decrease of the carrying angle as a function of increasing elbow flexion. After assessment of the carrying angle with a protractor goniometer and an electromagnetic tracking system (Flock of Birds) in extension, flexion-extension movements with the forearm held in supination were recorded by means of the latter system. Three recordings were averaged in both the left and the right elbows of 20 volunteers without a history of elbow pathology (10 males and 10 females; mean age 25 years). In extension, a mean (+/- SD) carrying angle of 11.6 +/- 3.2 degrees was found in the male and 16.7 +/- 2.6 degrees in the female subjects. The carrying angles progressively decreased with flexion, at the end changing into a mean (+/- SD) varus angle of 1.8 +/- 2.9 degrees in men and 1.6 +/- 2.3 degrees in women. Significant differences in carrying angles between the sexes were recorded in moving from 0 to 30 degrees of flexion (p < 0.03 for the left and p < 0.01 for the right elbows), but disappeared beyond 30 degrees . No statistically significant differences were found between the results of left and right elbows. Although statistically significant differences (p < 0.05 to p < 0.001) were found along the course of flexion and extension, these differences were small (<0.6 degrees ). The mean carrying angles at 0, 30, 60, 90 and 120 degrees of flexion revealed larger standard deviations in the male group than in the female group. PMID- 16338731 TI - Intra-articular kinematics of the upper limb joints: a six degrees of freedom study of coupled motions. AB - In sport, leisure and certain occupational activities, joint lesions of the upper limb are very common. To understand their aetiology in order to prevent and treat these pathologies on a scientific basis, a comprehensive knowledge of the involved stress and kinematics is imperative. For many years intra-articular joint kinematics have been described hypothetically in terms of the convex concave principle. This principle, however, has not been validated. The in vitro research on the acromioclavicular, glenohumeral and elbow joints was performed using an electromagnetic tracking device (Flock of Birds). By recalculating the positions of the trackers to an embedded coordinate system on the joint surface, the intra-articular joint movements can be analysed. Therefore, the bony configurations and articular surface features were registered with a 3D drawing stylus (Microscribe). Thirteen acromioclavicular joints, six glenohumeral joints and seven elbows were studied. The coupled intra-articular movements were analysed using the Euler angles and finite helical axis approaches. The results of the acromioclavicular joint analysis indicate that during movements in the scapular plane a superior rotation of the clavicle was coupled with an inferior translation and vice versa, whilst during movements in the plane perpendicular to the scapular plane the anterior rotation was coupled with an anterior translation and vice versa. In the glenohumeral joint, the abduction-elevation was coupled with an external rotation. In the humero-ulnar joint, the extension was coupled with an external rotation and varus movement. This intra-articular behaviour was in contradiction to currently accepted convex-concave concepts in arthrokinematics and manual therapy. The results may have major implications for manual therapy and orthopaedic medical practice. PMID- 16338732 TI - Changes in coordination, control and outcome as a result of extended practice on a novel motor skill. AB - It has been proposed that, when learning a motor skill, individuals initially freeze degrees of freedom to simplify control. There is limited empirical evidence to support this proposition. We examined this issue by monitoring the performance of a non-skilled individual learning a soccer chip shot with his non dominant leg over 9 days of practice (425 trials). Principal component analysis was used to examine dimensional change. The most dramatic change occurred at the hip, with the range of motion decreasing during the first 5 days of practice and then increasing thereafter. A reverse pattern was observed at the knee and ankle. While showing a progression in control from proximal to distal, a further phase was observed where primary control was passed back to the hip. The degree of linear coupling between the joints also increased with practice until day 5, after which independent control was observed. The number of controlled dimensions did not change across practice. Radial error decreased over practice and kinematics relating to the hip were most predictive of error, especially early in practice. Freezing degrees of freedom was a strategy implemented across the first half of practice, after which point-independent control was gradually restored enabling successful consistent performance. PMID- 16338733 TI - Anticipation and visual search behaviour in expert soccer goalkeepers. AB - A novel methodological approach is presented to examine the visual search behaviours employed by expert goalkeepers during simulated penalty kick situations in soccer. Expert soccer goalkeepers were classified as successful or unsuccessful based on their performance on a film-based test of anticipation skill, thereby allowing an intra-group comparison of visual search behaviour on the task. The anticipation test required participants to move a joystick in response to penalty kick situations presented on a large screen. The proportion of penalties saved was assessed as well as the frequency and time of initiation of joystick corrections. Visual search behaviour was examined using a portable eye movement registration system. The successful experts were more accurate in predicting the height and direction of the penalty kick, waited longer before initiating a response and appeared to spend longer periods of time fixating on the non-kicking leg compared with the non-successful experts. PMID- 16338734 TI - Multi-dimensional assessment of soccer coaching course effectiveness. AB - The purpose of this study was to determine the relationship between the aims of course providers and events during the delivery of two soccer coaching accreditation courses. A secondary purpose was to evaluate performance-analysis methods for assessing the course instructor's performance. A case analysis approach was developed to evaluate the courses and the data-gathering process. This research approach was chosen to amalgamate the sources of evidence, providing a multi-dimensional view of course delivery. Data collection methods included simple hand notation and computer logging of events, together with video analysis. The hand notation and video analysis were employed for the first course with the hand notation being replaced with computer event logging for the second course. Questionnaires, focusing on course quality, were administered to participants. Interviews and document analysis provided the researchers with the instructors' main aims and priorities for course delivery. Results of the video analysis suggest a difference between these aims and the events of the courses. Analysis of the questionnaires indicated favourable perceptions of course content and delivery. This evidence is discussed in relation to intent and practice in coach education and the efficiency of employing performance-analysis techniques in logging instructional events. PMID- 16338735 TI - Towards the reflective sports coach: issues of context, education and application. AB - The current demands faced by elite sports coaches and the nature of their educational experiences are outlined. Examination of current approaches to coach education are considered in relation to research evidence which describes the key skills required in elite sports coaching. Recently, reflective practice has emerged as a key skill with which to enhance coach learning and increase the value of coaches' educational experiences. In tracing this emergence, this study analyses the use of reflective practice and learning strategies within six governing body award coaching programmes. Results indicate that none of the programmes examined contained structures or processes for directly teaching or overtly nurturing reflective skills. However, almost all programmes offered a potential structure for this through completion of a coaching log or undertaking a period of mentoring. Finally, recommendations are made to coach educators for developing reflection within coach education programme structures. PMID- 16338737 TI - Better late than never: reflections on the delayed prioritization of cervical cancer in international health. AB - Cervical cancer has disproportionately affected the world's most vulnerable women for generations, yet only recently has the disease become an international health priority. Using ethnographic evidence from a study in Iquitos, Peru, I identify three factors that have contributed to a historic blind spot regarding cervical cancer in underdeveloped regions: (a) transition theories predicting the emergence of cancer with increasing societal development, (b) chronic vs. infectious disease categories that miss the significance of infectious-associated cancers, and (c) dependence on epidemiologic statistics from underdeveloped regions for determining health care priorities. Implications for theory and education are discussed. PMID- 16338738 TI - Transforming desolation into consolation: being a mother with life-threatening breast cancer. AB - The purpose of this study is to describe lived experiences of being ill with breast cancer for mothers with dependent children. A special focus is to explore meanings of desolation and consolation, and meanings of transforming the consolation. Stories of Swedish women who took part in a supportive network for young women with breast cancer were interpreted phenomenological-hermeneutically as transforming desolation into consolation. This means a changed direction in the longing and desires of the woman from outward to inward, from others to self, from emphasis on past and delimited presence to presence and future, and ending up in balancing between these opposites. Implications for women with breast cancer and their need for support are reflected upon. PMID- 16338739 TI - Factors related to depressive symptoms among long-term survivors of cervical cancer. AB - In this article we describe depressive symptoms among survivors of cervical cancer. Study participants were previously diagnosed and treated with cervical cancer 5 to 25 years prior to the completion of a telephone survey. Through a population-based survey of women identified via a state tumor registry in southern New England, the investigators learned that depressive symptoms are prevalent among a subgroup of long-term survivors long after diagnosis and treatment. The at-risk participants included those who were at a lower economic status and whose marital status and ability to work had changed, and who continued to experience pain and diarrhea caused by radiotherapy. Findings from this study warrant additional research with this potentially vulnerable group of women. PMID- 16338740 TI - Effects of breast cancer risk counseling for sexual minority women. AB - Sexual minority women (lesbian and bisexual) represent a vulnerable group regarding their breast health. The participants in this study were 150 women aged 18-74 recruited via public announcements in mainstream and sexual minority communities in the greater Seattle metropolitan area. Potential participants were recruited to participate in a randomized trial of a breast cancer risk counseling intervention for sexual minority women. The counseling intervention produced significant reductions in perceived risk of breast cancer, anxieties and fears about breast cancer at 6 and 24 months, and increases in breast screening rates at 24 months in the intervention arm, compared with the control arm participants. These data add to the growing body of knowledge on sexual minority women's health and point to areas of community action and future research. PMID- 16338741 TI - Comfort, control, or conformity: women who choose breast reconstruction following mastectomy. AB - Following breast amputation women commonly are presented with two choices: to wear a prosthesis or undergo reconstruction. Breast restoration is assumed to allow a full emotional and physical recovery from a breast cancer crisis. Surgical reconstruction is offered to women as the final step in regaining a sense of complete womanhood, enabling a sense of optimism that both body and self will "get back to normal." This article examines 5 women's accounts of breast reconstruction and asks how breast reconstruction figures in the remaking of self following mastectomy. Issues pertaining to the reasoning behind seeking out the procedure, experiences of finding the right surgeon, and how women feel toward their reconstructed postsurgical body are examined. In conclusion it is argued that a number of contradictory expectations are held by women seeking reconstructions. While women suggest that reconstruction will restore lost femininity, sexuality, and normalcy in most cases it is not the procedure that enables this but the elimination of the hassles of prostheses. In contrast to the complete sense of self they expected to regain through reconstruction they articulate a restoration that is simply pragmatic. It is only once women have undertaken this last bastion of hope that they are forced to renegotiate their sense of themselves as women with or without breasts. PMID- 16338742 TI - A qualitative analysis of reproductive issues raised by young Australian women with breast cancer. AB - Available literature concentrates on infertility concerns of young women with breast cancer, while attention to psychosocial issues related to maintained or regained fertility is scarce. As part of a longitudinal, qualitative study of experiences of young women with breast cancer (diagnosed at 40 years or younger); (n = 13), concerns about fertility, contraception, pregnancy, and breastfeeding after breast cancer were expressed. Using semistructured, one-to-one interviews over three time phases, we explored these women's experiences. Perceptions of fertility changed over time. Contraception issues were raised together with recurrence fears related to pregnancy and breastfeeding after breast cancer. Decisions related to unplanned pregnancies and breastfeeding were described as particularly onerous. PMID- 16338743 TI - Critical implication of the (70-96) domain of human immunodeficiency virus type 1 Vpr protein in apoptosis of primary rat cortical and striatal neurons. AB - The human immunodeficiency virus (HIV)-1 regulatory protein Vpr has been detected in the serum of HIV-seropositive individuals and in the cerebrospinal fluid of acquired immunodeficiency syndrome (AIDS) patients suffering from neurological disorders. Therefore, Vpr could play a critical role in the neuronal apoptosis observed postmortem in the brain of patients, often connected to a severe AIDS related disease termed HIV-associated dementia (HAD). This suggests that the Vpr neurotoxicity already observed in vitro on hippocampal neurons could also occur in other brain structures. In this study the authors have investigated the ability of synthetic Vpr to induce apoptosis in primary cultures of rat cortical and striatal neurons. Moreover, the authors have explored the Vpr minimal proapoptotic region using synthetic Vpr fragments and mutants of the protein. Treatments of both neuronal types with Vpr, its C-terminal domain, Vpr(52-96), or a shorter fragment, Vpr(70-96), led to dose- and time-dependent cell death as determined by flow cytometry after propidium iodide labeling, phase-contrast microscopy, and TUNEL labeling. Taken together, these results support an apoptosis-induced death of these neurons. The (71-82) Vpr peptide, previously shown toxic to isolated mitochondria, was inactive on neurons. Vpr-induced neuronal apoptosis was associated with activation of caspase-3 beginning 3 h after Vpr extracellular addition and peaking 3 h later. Moreover, an hyperproduction of reactive oxygen species was observed. In addition to hippocampal neurons, the extension of the apoptotic property of Vpr to cortical and striatal neurons could account for several signs observed in HAD and is thus consistent with a possible involvement of Vpr in this syndrome. PMID- 16338744 TI - Clinical validation of the NeuroScreen. AB - The NeuroScreen comprises two easily administered components: the Brief NeuroCognitive Screen (BNCS), designed to estimate the frequency of human immunodeficiency virus (HIV)-associated cognitive disorders; and the Brief Peripheral Neuropathy Screen (BPNS), for distal sensory polyneuropathy (DSPN) in HIV. In this study, both the NeuroScreen and a more extensive standardized validation neurodiagnostic evaluation were administered to HIV-positive subjects (N = 301) enrolled in two large cohort studies at multiple sites. BNCS performance was summarized in the form of a demographically adjusted mean z score, the NPZ3. The area under the receiver-operating characteristic (ROC) curve for the BNCS as compared to the reference standard neuropsychological (NP) evaluation was 0.74 (95% confidence interval [CI] 0.69, 0.79). Using a cut-point of -0.33 on the NPZ3 provided a correct classification rate of 68%, with roughly balanced sensitivity (65%) and specificity (72%). Under the assumption of a 30% prevalence of cognitive impairment, the calculated positive predictive value (PPV) of the BNCS was 86%. Relative to its reference standard, a modified Total Neuropathy Score (TNS) administered by a neurologist, the BPNS gave a similar correct diagnostic classification rate of 78%; sensitivity 49% [95%, 60%]; specificity 88% [95% Cl82%, 91%]. Under the assumption of a 40% prevalence of DSPN, the PPV of the BPNS was 72%. These predictive values suggest that the NeuroScreen will be useful for tracking trends in the prevalence of HIV associated neurologic disease in large cohorts in the era of combination antiretroviral therapy. However, because it yields substantial numbers of false positives and negatives, the NeuroScreen may be less useful in evaluating individual patients. PMID- 16338745 TI - Differential responses of human brain cells to West Nile virus infection. AB - In recent years, West Nile virus (WNV) has emerged as a major cause of encephalitis in the United States. However, the neuropathogenesis of this flavivirus is poorly understood. In the present study, the authors used primary human brain cell cultures to investigate two neuropathogenic features: viral replication and induction of cytokines. Although neurons and astrocytes were found to support productive WNV infection, viral growth was poorly permissive in microglial cells. Compared to neuronal cultures that sustained viral growth for at least 2 weeks, replication peaked in astrocytes by 72 h post infection. In response to viral infection, astrocytes produced chemokines (CXCL10 and CCL5), but none of the cytokines (tumor necrosis factor [TNF]-alpha, interleukin [IL] 1beta, IL-6, interferon alpha or gamma) tested could be detected. Although microglial cells failed to support viral replication, WNV induced production of the proinflammatory cytokines IL-6 and TNF-alpha. Microglial cells also released robust amounts of the chemokines CXCL10 and CCL2, as well as lower levels of CCL5, in response to WNV infection. WNV-induced chemokine and cytokine production by microglia was coupled with activation of mitogen-activated protein kinase (MAPK) intracellular signaling pathways. Inhibition of p38 MAPK decreased chemokine production in response to WNV. Taken together, these findings suggest that microglial cell responses may influence the neuropathogenesis of WNV infection. PMID- 16338746 TI - Conventional magnetic resonance imaging features in patients with tropical spastic paraparesis. AB - Conventional brain and spinal cord magnetic resonance images were performed in 21 patients with human T-cell lymphotropic virus (HTLV)-1 associated myelopathy/tropical spastic paraparesis, to assess the role of conventional magnetic resonance imaging (MRI) in the disease diagnosis. These patients had no other central nervous system conditions or related risk factors at the time of tropical spastic paraparesis diagnosis. Eleven (52.4%) patients showed nonspecific brain abnormalities on T2-weighted images. The majority (77.2%) of brain abnormalities were located in the deep white matter. A transient contrast enhancing lesion was identified in the brain of only one patient. In the brain of another patient, 9.0% of the T2-hyperintense lesion load was hypointense on the correspondent T1-weighted images. No differences in terms of demographic, biological, or clinical variables were present between patients with abnormal brain images and those with normal brain magnetic resonance images. Spinal cord T2-weighted images were abnormal in three (14.3%) patients. In one of these three patients, a diffuse but transient edema was found along the entire tract of the spinal cord. White matter lesions were present in the central nervous system of 60% of the cases in this study. However, no correlations between magnetic resonance imaging and clinical findings, and no specificity of lesions were observed. Hence, conventional magnetic resonance imaging is a sensitive but not highly specific tool for diagnosis of tropical spastic paraparesis. PMID- 16338747 TI - Aquaporin 4 is increased in association with human immunodeficiency virus dementia: implications for disease pathogenesis. AB - Changes in astrocyte shape and function are known to occur in association with human immunodeficiency virus (HIV) dementia (HIVD). However, the causes and consequences of such changes are not completely understood. In vitro data suggest that changes in the expression of aquaporin 4 (AQP4), the aquaporin subtype expressed by astrocytes, can significantly influence cell shape and physiology. In the present study, the authors therefore investigated the possibility that AQP4 levels may be altered in HIVD. Using Western blot, the authors show that immunoreactivity for AQP4 is elevated in brain homogenates from the mid frontal gyrus of patients who died with HIVD (P < .005 HIV seronegative versus HIVD). Of interest, a significant increase was also observed in homogenates from HIV infected individuals without dementia (P < .05 HIV seronegative versus neurologically normal HIV seropositive). In the present study the authors also examined the stimulated expression of AQP4 in cultured cells. Previous in vitro studies have shown that AQP4 expression may be increased by stimuli that induce cytoskeletal changes and/or the activation of p38 mitogen-activated protein (MAP) kinase. The authors therefore focused on tumor necrosis factor (TNF)-alpha, which has been linked to p38 MAP kinase activation, and thrombin, which may also induce changes in the actin cytoskeleton. Both may be elevated with HIVD. Again using Western blot, the authors show an increase in both AQP4 and phosphorylated p38 MAP kinase in homogenates from TNF-alpha- and thrombin-stimulated organotypic cerebellar and spinal cord cultures. Together, these studies suggest that AQP4 expression may be altered in HIVD and/or in response to exogenous proteinases. Additional studies may be warranted to determine whether altered AQP4 expression represents a protective and/or maladaptive response to central nervous system (CNS) inflammation. PMID- 16338748 TI - Expression and evolutionary analysis of West Nile virus (Merion strain). AB - The authors report a new strain of West Nile virus (WNV) with the expression analysis of its individual open reading frames. Since its sudden appearance in the summer of 1999 in New York City, the virus has spread rapidly across the continental United States into Canada and Mexico. Besides, its rapid transmission by various vectors, the spread of this virus through organ transplantation, blood transfusion, and mother-child transmission through breast milk is of concern. In order to understand molecular variations of WNV in North America and to generate new tools for understanding WNV biology, a complete clone of WNV has been constructed. Investigations so far have focused only on half of its genes products and a detailed molecular and cell biological aspects on all of WNV gene have yet to be clearly established. The open reading frames of WNV were recovered through an reverse transcriptase-polymerase chain reaction (RT-PCR)-PCR using brain tissue from a dead crow collected in Merion, PA, and cloned into a mammalian expression vector. The deduced amino acid sequences of individual open reading frames were analyzed to determine various structural motifs and functional domains. Expression analysis shows that in neuronal cells, C, NS1, and NS5 proteins are nuclear localized whereas the rest of the antigens are confined to the cytoplasm when they are expressed in the absence of other viral antigens. This is the first report that provides an expression analysis as well as intracellular distribution pattern for all of WNV gene products, cloned from an infected bird. Evolutionary analysis of Merion strain sequences indicates that this strain is distinct phylogenetically from the previously reported WNV strains. PMID- 16338749 TI - Cerebrospinal fluid proteomics and human immunodeficiency virus dementia: preliminary observations. AB - Protein profiling using mass spectrometry may be useful in identifying previously unknown protein markers in human immunodeficiency virus (HIV) dementia and provide insight into disease pathogenesis. Six samples of matched cerebrospinal (CSF) and blood serum from patients with no, mild, and moderate dementia were prepped for biomarker screening by the Ciphergen system. Chips were analyzed in the matrix-assisted laser desorption/ionization (MALDI) mass spectrometer at low mass (700 to 20,000 Da) and at higher mass (5000 to 100,000 Da). In both serum and CSF samples, differences in protein intensity appeared to correlate with degree of dementia. This preliminary study suggests that protein markers of HIV dementia may be detected by MALDI mass spectrometry. PMID- 16338750 TI - Localization of sequences within the latency-related gene of bovine herpesvirus 1 that inhibit mammalian cell growth. AB - The latency-related (LR) RNA of bovine herpes virus 1 (BHV-1) is abundantly expressed in sensory neurons of latently infected cattle. Wild-type expression of LR gene products is required for the latency-reactivation cycle. LR gene products inhibit apoptosis, bICP0 expression, and mammalian cell growth. The cell growth inhibitory function of the LR gene maps to a 463-bp XbaI-PstI fragment. Introduction of stop codons into the XbaI-PstI fragment had no effect on inhibiting growth. Expression of a LR strand-specific transcript correlates with growth inhibition in bovine fibroblasts and mouse neuroblastoma cells. PMID- 16338751 TI - Glycoprotein-mediated induction of apoptosis limits the spread of attenuated rabies viruses in the central nervous system of mice. AB - Induction of apoptosis by rabies virus (RV) has been reported to be associated with the expression of the glycoprotein (G), but inversely correlated with pathogenicity. To further delineate the association between the expression of the G and the induction of apoptosis, recombinant RVs with replacement of only the G gene were used to infect mice by the intracerebral route. Recombinant viruses expressing the G from attenuated viruses expressed higher level of the G and induced more apoptosis in mice than recombinant RV expressing the G from wild type (wt) or pathogenic RV, demonstrating that it is the G gene that determines the level of G expression and, consequently, the induction of apoptosis. Likewise, recombinant viruses expressing the G from wt or pathogenic RV are more pathogenic in mice than those expressing G from attenuated RV, confirming the inverse correlation between RV pathogenicity and the induction of apoptosis. To investigate the mechanism by which induction of apoptosis attenuates viral pathogenicity, mice were infected with wt or attenuated RV by the intramuscular route. It was found that low doses of attenuated RV induced apoptosis in the spinal cord and failed to spread to the brain or produce neurological disease. On the other hand, apoptosis was not observed in the spinal cord of mice infected with the same doses of wt RV and the virus spread to various parts of the brain and induced fatal neurologic disease. These results suggest that glycoprotein mediated induction of apoptosis limits the spread of attenuated rabies viruses in the central nervous system (CNS) of mice. PMID- 16338752 TI - Ten-year changes in cancer-related health behaviors and screening practices among Latino women and men in California. AB - OBJECTIVE: This study examines changes in cancer-related health behaviors and risk factors (overweight/obesity, unhealthy diet, high alcohol use, and smoking), and screening practices related to cervical, breast, and colorectal cancer among Latinos of predominantly Mexican origin in Monterey County, California. DESIGN: Data is from two cross-sectional surveys, conducted in 1990 and 2000, that included 919 women and 774 men from a community sample, and 276 men from an agricultural labor camp sample (ages 18-64). RESULTS: Over the 10-year period, the prevalence of obesity increased by 48% among community women, 47% among community men, and 91% among labor camp men. Although consumption of fruits and vegetables remained low and consumption of fried foods remained high, other diet related behaviors showed significant improvements (e.g. milk consumption shifted from whole-fat to lower-fat among women from the community and men from the labor camps, use of lard or meat fat when cooking decreased among women and men from the community). In addition, alcohol intake decreased among men from both samples, as did smoking among labor camp men. There were large improvements for annual pap and mammography screening (increases from 53 to 71% for pap testing, and from 15 to 53% for mammography screening) but annual blood stool testing remained infrequent and unchanged. CONCLUSION: These findings highlight the need for interventions and policies that improve knowledge, preventive care, and social environments to sustain improvements and address areas of special need in cancer prevention for Latinos, especially related to obesity and colorectal screening. PMID- 16338753 TI - Attitudes, knowledge, and health-seeking behaviors of five immigrant minority communities in the prevention and screening of cancer: a focus group approach. AB - Despite an observed decrease in overall cancer death rates in the USA, immigrant minorities continue to experience disproportionately higher cancer incidence and mortality rates. Thirteen focus groups were conducted in the Haitian, English Speaking Caribbean, Latino, Korean, and Chinese communities of New York City to better understand their health-seeking behaviors with respect to cancer prevention, screening, and treatment. Focus groups addressed the degree to which cultural, linguistic, and systematic barriers impact these behaviors and explored methods to support salutary behaviors. Findings underscored that, while there are many similarities across immigrant groups, there are significant variations between the immigrant groups to necessitate a tailored community-based approach. The prevalent misinformation observed among all groups warrants the prompt development of culturally competent programs for cancer control with immigrant minorities. PMID- 16338754 TI - Caregiving outside of the home: the effects of race on depression. AB - OBJECTIVE: This research examines the conditions that determine whether Blacks experience lower or higher levels of depression while caregiving outside of the home, as compared to Whites. Some prior literature has found that African Americans report a lesser caregiver burden despite an increased likelihood that they will acquire this role, and decreased resources to do so. Others have found that African Americans experience the same caregiver burden and distress as Whites. Given these mixed findings, we use the stress process model to examine whether African American caregivers experience lower or higher levels of depression when they provide care outside of the home. DESIGN: A sample of care workers who provide care to others outside of the home was drawn from the 1992-4 National Survey of Families and Households. The final sample included 275 (11%) Blacks, and 2,218 (89%) Whites (not of Hispanic origin). The primary statistical method for predicting differences in caregivers' depressive symptomatology was OLS regression analysis with progressive adjustment. RESULTS: We examined sociodemographics, family structure, resources, and stressors and found that African Americans, those with lower socioeconomic status, the unmarried, spending more weeks caregiving, having a physical impairment, and surprisingly receiving more help from parents are associated with higher depressive symptomatology. Stronger religious beliefs decreased depressive symptomatology for Blacks. The race effect was, in part, explained by family structure, amount of caregiving, and impairment of care worker. CONCLUSION: Contrary to prior literature, we found that Blacks are more depressed than White caregivers in large part because of lower socioeconomic status and greater stressors, and higher levels of physical impairment. Yet, strength in religious belief has a stress-buffering effect for African Americans. We suggest that policies that attempt to eliminate racial disparities in socioeconomic status and health could benefit these caregivers. PMID- 16338755 TI - Ethnic differences in the relationship between depressive symptoms and health related quality of life in people with type 2 diabetes. AB - OBJECTIVE: To examine ethnic differences in the relationship between depressive symptoms and specific aspects of heath-related quality of life in people with type 2 diabetes. DESIGN: Cross-sectional data from 190 people with type 2 diabetes of Native Hawaiian (50%), Filipino (16%), Japanese (18%), and mixed ethnic (16%) ancestries from the rural community of North Kohala, Hawai'i were examined in this study. Depressive symptoms were measured with the Center for Epidemiological Studies-Depression (CES-D) scale. Health-related quality of life was measured with the Short Form-36 Health Survey (SF-36). Eight health-related quality of life constructs were examined: Physical Functioning, Role-Physical Functioning, Role-Emotional Functioning, Social Functioning, Bodily Pain, Vitality, General Health, and Health Transition. RESULTS: Hierarchical regression analyses of the interaction between ethnicity and the SF-36 subscales of Physical Functioning, Role-Emotional Functioning, Bodily Pain, Vitality, and General Health indicated statistically significant associations with CES-D scores after controlling for sociodemographic factors, glycemic status, and social support. CONCLUSION: Ethnicity moderated the relationship between depressive symptoms and the health-related quality of life aspects of physical and role-emotional functioning, bodily pain, vitality, and general health perception in people with type 2 diabetes. This relationship was strongest for Filipinos followed by Native Hawaiians and people of mixed-ethnic ancestries. PMID- 16338756 TI - The role of intimate partner violence, race, and ethnicity in help-seeking behaviors. AB - OBJECTIVE: Women experiencing intimate partner violence (IPV) have multiple health and social service needs but many, especially Hispanic, women may not access these resources. This research sought to examine the relationship between IPV and health and social services utilization (help-seeking behaviors), with a focus on racial and ethnic disparities. DESIGN: Case-control study from an urban US emergency department population in which cases (women with IPV) and controls (women without IPV) were frequency matched by age group and race/ethnicity. Logistic regression analyses were performed to examine the relationship between IPV and help-seeking behaviors and between help-seeking behaviors and race/ethnicity among abused women. In addition, a stratified analysis was conducted to examine the relationship between acculturation and help-seeking behaviors among Hispanic women. RESULTS: The sample included 182 cases and 147 controls. Among the health services, alcohol program, emergency department, and hospital utilization were significantly increased among IPV victims compared to non-victims after taking demographic and substance use factors into account. Similarly, IPV victims were more likely to access social/case worker services and housing assistance compared to non-victims. Specific help-seeking behaviors were significantly associated with race and ethnicity among IPV victims, with non Hispanic white and black women more likely to use housing assistance and emergency department services and black women more likely to use police assistance compared to Hispanic women. Among all Hispanic women, low acculturation was associated with decreased utilization of social services overall and with any healthcare utilization, particularly among abused women. CONCLUSIONS: Social service and healthcare workers should be alerted to and screen for IPV among all clients. The need for increased outreach and accessibility of services for abused women in Hispanic communities in the USA should be addressed, with cultural and language relevance a key component of these efforts. PMID- 16338757 TI - Polycystic liver and kidney diseases. AB - There have been remarkable advances in research on polycystic liver and kidney diseases recently, covering cloning of new genes, refining disease classifications, and advances in understanding more about the molecular pathology of these diseases. Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary disease affecting kidneys. It affects 1/400 to 1/1000 live births and accounts for 5% of the end stage renal disease in the United States and Europe, and is caused by gene defects in the PKD1 or PKD2 genes. Compared to ADPKD, polycystic liver disease (PCLD) is a milder disease and does not lower life expectancy. Both diseases are usually adult-onset diseases. Defects in genes, which code the hepatocystin and SEC63 proteins, have just recently been found to cause PCLD. It now seems that ADPKD is caused by malfunction of the primary cilia, a cell organ sensing fluid movement, and that PCLD is a sequel from defects in protein processing. Autosomal recessive polycystic kidney disease (ARPKD) belongs to a group of congenital hepatorenal fibrocystic syndromes. All ARPKD patients have a gene defect in a gene called PKHD1, the protein product of which localizes to primary cilia. We summarize the present clinical and molecular knowledge of these diseases in this review. PMID- 16338758 TI - Inhibition of Nogo: a key strategy to increase regeneration, plasticity and functional recovery of the lesioned central nervous system. AB - In the adult central nervous system (CNS) myelin and oligodendrocytes, Nogo-A exerts a growth inhibitory function leading to restricted axonal regeneration. After development of different anti-Nogo-A antibodies and other Nogo-A blocking reagents their application has recently been studied in various in vivo animal models of spinal cord injury and stroke. These studies show that intracerebral application of Nogo-A-inactivating reagents leads to enhanced regeneration and compensatory sprouting, structural reorganization or plasticity, and functional recovery as seen in different behavioural analyses. PMID- 16338759 TI - Epidemiology and diagnosis of Lyme borreliosis. AB - The multisystem disease Lyme borreliosis is the most frequent tick-transmitted disease in the northern hemisphere. In Europe Lyme borreliosis is most frequent in Central Europe and Scandinavia (up to 155 cases per 100,000 individuals) and is caused by the species, B. burgdorferi sensu stricto, B. afzelii and B. garinii. The recently detected genospecies A14S may also play a role in skin manifestations. Microbiological diagnosis in European patients must consider the heterogeneity of borreliae for development of diagnostic tools. According to guidelines of the USA and Germany, serological diagnosis should follow the principle of a two-step procedure (enzyme-linked immunosorbent assay (ELISA) as first step, if reactive; followed by immunoblot). The sensitivity and standardization of immunoblots has been considerably enhanced by use of recombinant antigens (p100, p58, p41i, VlsE, OspC, DbpA) including those expressed primarily in vivo (VlsE and DbpA) instead of whole cell lysates. VlsE is the most sensitive antigen for IgG antibody detection, OspC for IgM antibody detection. At present, detection rates for serum antibodies are 20%-50% in stage I, 70%-90% in stage II, and nearly 100% in stage III Lyme disease. Detection of the etiological agent by culture or polymerase chain reaction (PCR) should be confined to specific indications and specialized laboratories. Recommended specimens are skin biopsy specimens, cerebrospinal fluid (CSF) and synovial fluid. The best results are obtained from skin biopsies with culture or PCR (50% 70%) and synovial tissue or fluid (50%-70% with PCR). CSF yields positive results in only 10%-30% of patients except when the duration of symptoms is shorter than 2 weeks (50% sensitivity). Methods which are not recommended or adequately documented for diagnosis are antigen tests on body fluids, PCR of urine, and lymphocyte transformation tests. PMID- 16338760 TI - Genes for normal sleep and sleep disorders. AB - Sleep and wakefulness are complex behaviors that are influenced by many genetic and environmental factors, which are beginning to be discovered. The contribution of genetic components to sleep disorders is also increasingly recognized as important. Point mutations in the prion protein, period 2, and the prepro hypocretin/orexin gene have been found as the cause of a few sleep disorders but the possibility that other gene defects may contribute to the pathophysiology of major sleep disorders is worth in-depth investigations. However, single gene disorders are rare and most common disorders are complex in terms of their genetic susceptibility, environmental effects, gene-gene, and gene-environment interactions. We review here the current progress in the genetics of normal and pathological sleep. PMID- 16338761 TI - Serotonin gene polymorphisms and bipolar I disorder: focus on the serotonin transporter. AB - The pathogenesis of bipolar disorder may involve, at least in part, aberrations in serotonergic neurotransmission. Hence, serotonergic genes are attractive targets for association studies of bipolar disorder. We have reviewed the literature in this field. It is difficult to synthesize results as only one polymorphism per gene was typically investigated in relatively small samples. Nevertheless, suggestive associations are available for the 5HT2A receptor and the serotonin transporter genes. With the availability of extensive polymorphism data and high throughput genotyping techniques, comprehensive evaluation of these genes using adequately powered samples is warranted. We also report on our investigations of the serotonin transporter, SLC6A4 (17q11.1-q12). An insertion/deletion polymorphism (5HTTLPR) in the promoter region of this gene has been investigated intensively. However, the results have been inconsistent. We reasoned that other polymorphism/s may contribute to the associations and the inconsistencies may be due to variations in linkage disequilibrium (LD) patterns between samples. Therefore, we conducted LD analyses, as well as association and linkage using 12 polymorphisms, including 5HTTLPR. We evaluated two samples. The first sample consisted of 135 US Caucasian nuclear families having a proband with bipolar I disorder (BDI, DSM IV criteria) and available parents. For case-control analyses, the patients from these families were compared with cord blood samples from local Caucasian live births (n = 182). Our second, independent sample was recruited through the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD, 545 cases, 548 controls). No significant associations were detected at the individual polymorphism or haplotype level using the case-control or family-based analyses. Our analyses do not support association between SLC6A4 and BDI families. Further studies using sub-groups of BDI are worthwhile. PMID- 16338762 TI - Relationship between tumor necrosis factor-alpha and ammonia in patients with hepatic encephalopathy due to chronic liver failure. AB - BACKGROUND: We have recently demonstrated that in humans, circulating levels of tumor necrosis factor-alpha (TNF) correlate positively with severity of hepatic encephalopathy (HE) due to chronic liver failure.AIM. The main aim of this larger population study is to determine the relationship between TNF and ammonia in patients with HE and chronic liver failure due to liver cirrhosis. METHODS: Circulating levels of TNF and ammonia were measured in 108 patients with liver cirrhosis due to various etiologies in various clinical grades of HE (grades 0 4). TNF concentrations were measured in venous serum using commercially available solid-phase high sensitivity enzyme-linked immunosorbent assay. Ammonia levels were determined in venous plasma by the enzymatic method, using the glutamate dehydrogenase reaction. RESULTS: The mean+/-SEM values of circulating levels of TNF and ammonia at presentation in patients with grade 0 of HE (n = 30) were 3.89+/-0.2 pg/mL and 49.8+/-2.8 microg/mL respectively, in patients with grade 1 of HE (n = 26) were 8.56+/-0.34 pg/mL and 101.6+/-6.5 microg/mL respectively, in patients with grade 2 of HE (n = 22) were 11.59+/-0.48 pg/mL and 160.3+/-10.7 microg/mL respectively, in patients with grade 3 of HE (n = 20) were 19.98+/-0.94 pg/mL and 228.8+/-16.1 microg/mL respectively, and in patients with grade 4 of HE (n = 10) were 51.53+/-8.59 pg/mL and 284.2+/-20.3 microg/mL respectively. A significant positive correlation was found between circulating levels of TNF and those of ammonia (r = 0.62, P< 0.0001), and also between circulating levels of both substances and severity of HE in these patients (r = 0.95, P<0.0001, and r = 0.9, P<0.0001 respectively). TNF and ammonia were both significant independent predictors of severity of HE (P<0.0001 for both variables). CONCLUSION: The results of this study demonstrate a significant relationship between TNF and ammonia in patients with chronic liver failure and HE, and so strengthen the suggestion that TNF could be strongly involved in the pathogenesis of HE in these patients. Hence, we suggest a new theory in the pathogenesis of HE, the "TNF theory". PMID- 16338763 TI - Parameters of antioxidative defense in type 2 diabetic patients with cardiovascular complications. AB - OBJECTIVE: Diabetes-associated oxidative stress is a consequence of both increased production of free radicals and reduced capacity of antioxidative defense. Prolonged hyperglycemia is the major factor in the pathogenesis of atherosclerosis in diabetes which can lead to cardiovascular complications. The aim of this study was to test the parameters of antioxidative defense in type 2 diabetic patients. METHODS: A total of 117 type 2 diabetics with and without cardiovascular complications were examined in order to find out the influence of hyperglycemia, type and duration of complications and duration of diabetes on the extent of disorder of antioxidative parameter values: superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and total antioxidant status (TAS). RESULTS: Compared to healthy control subjects, type 2 diabetic patients with cardiovascular complications (CVC) had significantly lower SOD (P<0.0001), GSH-Px (P<0.0001), GR ( P = 0.0002) and TAS values (P<0.0001). In type 2 diabetic subjects with CVC, males had significantly lower SOD (778.7+/ 103.2 U/gHb, P<0.01) and GR activities (52.2+/-8.9 U/L, P<0.001) compared to females (839.3+/-94.9 U/gHb; 58.5+/-9.1 U/L). Significant and positive correlation was found between glucose levels and SOD (r = 0.375 for P<0.05) and GSH-Px (r = 0.384, P<0.05 ) activity in the group of complications-free diabetics, while significant negative correlation between glucose and GSH-Px values (r = -0.382, P<0.05) was found in the group of type 2 diabetics with coronary artery disease (CAD) and hypertension (HTA) and with CAD and acute myocardial infarction (AMI) (r = -0.860 P<0.05), and highly negative correlation between glucose and SOD levels (r = -0.590, P<0.05) in the group of diabetic subjects with CAD, AMI and HTA. Likewise, there was highly significant negative correlation of SOD (r = -0.949, P<0.05) and TAS (r = -0.393 for P = 0.038) with duration of diabetes in the group of diabetics with CAD and HTA. CONCLUSION: Our results confirm the hypothesis that there is reduced antioxidative defense in type 2 diabetics with prominent cardiovascular complications, which negatively correlates with glucose concentrations and duration of diabetes and cardiovascular complications. PMID- 16338764 TI - Molecular phylogenetics of the siphonophora (Cnidaria), with implications for the evolution of functional specialization. AB - Siphonophores are a group of pelagic colonial hydrozoans (Cnidaria) that have long been of general interest because of the division of labor between the polyps and medusae that make up these "superorganisms." These polyps and medusae are each homologous to free living animals but are generated by an incomplete asexual budding process that leaves them physiologically integrated. They are functionally specialized for different tasks and are precisely organized within each colony. The number of functional types of polyps and medusae varies across taxa, and different authors have used this character to construct phylogenies polarized in opposite directions, depending on whether they thought siphonophore evolution proceeded by a reduction or an increase in functional specialization. We have collected taxa across all major groups of siphonophores, many of which are found exclusively in the deep sea, using remotely operated underwater vehicles (ROVs) and by SCUBA diving from ships in the open ocean. We have used 52 siphonophores and four outgroup taxa to estimate the siphonophore phylogeny with molecular data from the nuclear small subunit ribosomal RNA gene (18S) and the mitochondrial large subunit ribosomal RNA gene (16S). Parsimony reconstructions indicate that functionally specialized polyps and medusae have been gained and lost across the phylogeny. Maximum likelihood and Bayesian analyses of morphological data suggest that the transition rate for decreased functional specialization is greater than the transition rate for increased functional specialization for three out of the four investigated categories of polyps and medusae. The present analysis also bears on several long-standing questions about siphonophore systematics. It indicates that the cystonects are sister to all other siphonophores, a group that we call the Codonophora. We also find that the Calycophorae are nested within the Physonectae, and that the Brachystelia, a historically recognized grouping of short-stemmed taxa, are polyphyletic. [Cnidaria; colonial animals; deep sea; division of labor; functional specialization; Hydrozoa; phylogenetics; Siphonophores.]. PMID- 16338765 TI - Unearthing the molecular phylodiversity of desert soil green algae (Chlorophyta). AB - Deserts are not usually considered biodiversity hotspots, but desert microbiotic crust communities exhibit a rich diversity of both eukaryotic and prokaryotic life forms. Like many communities dominated by microscopic organisms, they defy characterization by traditional species-counting approaches to assessing biodiversity. Here we use exclusive molecular phylodiversity (E) to quantify the amount of evolutionary divergence unique to desert-dwelling green algae (Chlorophyta) in microbiotic crust communities. Given a phylogenetic tree with branch lengths expressed in units of expected substitutions per site, E is the total length of all tree segments representing exclusively desert lineages. Using MCMC to integrate over tree topologies and branch lengths provides 95% Bayesian credible intervals for phylodiversity measures. We found substantial exclusive molecular phylodiversity based on 18S rDNA data, showing that desert lineages are distantly related to their nearest aquatic relatives. Our results challenge conventional wisdom, which holds that there was a single origin of terrestrial green plants and that green algae are merely incidental visitors rather than indigenous components of desert communities. We identify examples of lineage diversification within deserts and at least 12 separate transitions from aquatic to terrestrial life apart from the most celebrated transition leading to the embryophyte land plants. [Bayesian phylogenetics; biodiversity; exclusive molecular phylodiversity; microbiotic crusts.]. PMID- 16338766 TI - Effects of treated and untreated depressive symptoms on highly active antiretroviral therapy use in a US multi-site cohort of HIV-positive women. AB - This study examines the effects of treated and untreated depressive symptoms on the likelihood of utilization of highly active antiretroviral therapy (HAART) among a multi-site cohort of HIV-infected women who screened positive for probable depression. Data were collected biannually from 1996 through 2001 in a prospective cohort study. Random-effects regression analysis was used to estimate the longitudinal effects of mental health treatment on the probability of HAART utilization, controlling for clinical indicators (CD4 count, viral load), demographic features (race/ethnicity, income), and behavioural factors (recent crack, cocaine, or heroin use). Use of antidepressants plus mental health therapy, or use of mental health therapy alone significantly increased the probability of HAART utilization, compared to receiving no depression treatment. Use of antidepressants alone did not differ significantly from receiving no depression treatment. African American women and those who used crack, cocaine, or heroin also were less likely to use HAART. These findings suggest that efforts to enhance depressed women's access to psychopharmacologic treatment and therapy may increase their use of the most effective HIV therapies. PMID- 16338767 TI - Drastic elevations in mortality among female injection drug users in a Canadian setting. AB - The health and social conditions of women living in Vancouver's Downtown Eastside has recently been the focus of substantial international attention. Since few studies have examined rates and correlates of death among addicted women in Canada, we have characterized patterns of mortality among female injection drug users (IDUs) in Vancouver. The Vancouver Injection Drug Users Study (VIDUS) is a prospective open cohort study of IDUs. The analyses presented here, were restricted to women enrolled between May 1996 and May 2002 and who were aged 14 years or older. We estimated cumulative mortality rates using Kaplan-Meier methods and Cox regression was used to calculate univariate and adjusted relative hazards. Between May 1996 and May 2002, 520 female IDUs have been recruited from the Vancouver area among whom 68 died during the study period. Elevated rates of mortality were observed among those who reported, baseline sex-trade involvement, those with HIV-infection at baseline, and those who lived in unstable housing at baseline (all log-rank: p<0.05). In adjusted analyses, HIV infection (RH = 3.09 [95% CI: 1.86-5.11]; p<0.001), unstable housing (RH = 1.74 [95% CI: 1.10-2.86]; p=0.029) and sex-trade involvement (RH = 1.82 [95% CI: 0.95-3.45]; p=0.071) were associated with the time to death. When the number of observed deaths was compared to the number of expected deaths based on the general female population of British Columbia using indirect standardization, the rate of death among female IDUs was elevated by a factor of 47.3 (95% CI: 36.1-58.5). In Vancouver, female IDUs have rates of mortality almost 50 times that of the province's female population. Our findings are consistent with a growing number of reports from other settings internationally, and demonstrate the need for an appropriate evidence-based strategy to address the health and social needs of addicted women. PMID- 16338768 TI - Effectiveness and cost effectiveness of early and late prevention of HIV/AIDS progression with antiretrovirals or antibiotics in Southern African adults. AB - As HIV/AIDS drugs are becoming more widely available in Southern Africa, we compared the effectiveness and cost effectiveness of different treatment options, using a Markov Monte Carlo simulation model based on published estimates of disease progression, treatment effectiveness and health care costs. Cost and outcome values were discounted. Quality of life was considered. Acceptability curves summarized uncertainties. Sensitivity analyses tested assumptions. Results showed that triple antiretroviral therapy (ARV) plus antibiotics would prolong life by 6.7 undiscounted years if provided 'late' (CD4 = 200 cells/microl) and by 9.8 years if provided 'early' (CD4 = 350 cells/microl). The incremental undiscounted costs per year of life gained, compared to no preventive therapy, were $17 for isoniazid plus cotrimoxazole started late, $244 for both antibiotics started early, $2454 for ARV plus antibiotics started late and $2784 for ARV plus both antibiotics started early. The discounted incremental costs per quality adjusted life year (QALY) gained were, respectively, $29 saving, $254, $4937 and $3057. Late ARV plus both antibiotics was the strategy most likely to be cost effective if society was willing to pay more than $2000 per life year gained. Cost-effectiveness estimates were sensitive to discounting and assumed treatment costs but were less sensitive to assumed treatment effectiveness. PMID- 16338769 TI - Costs and financial burden of care and support services to PLHA and households in South India. AB - The objective of this study is to estimate the medical and non-medical out-of pocket expenditure on care and support services to PLHA, the financial burden on households, the indirect costs and coping strategies to meet the financial burden. A structured pre-tested questionnaire was used to collect data from a cohort of 153 clients of YRG CARE, a leading Chennai based NGO, who had completed the first and third waves of interview during 2000/01 and 2001/02 respectively. The results show that the median out-of-pocket medical and non-medical expenditures (direct cost) for treatment and services are Rs. 6,000 (US $ 122) in a reference period of six months. Clients on antiretroviral (ARV) drugs spend five times more than those not on ARV. The median direct cost significantly increases with stage of disease, household income, and poverty level. The financial burden of treatment, measured as the ratio of direct cost to household income, is greater on lower income (82%) than on higher income (28%) households. 31% and 45% of the clients reported loss of income and workdays respectively. In conclusion, the direct costs and financial burden of care and support services increase with the stage of disease. The financial burden is disproportionately more on low-income households. HIV/AIDS leads to depletion of savings and increases the indebtedness of households. PMID- 16338770 TI - Perceived social environment and quality of life among people living with HIV/AIDS in northern Thailand. AB - Efforts have been made to improve the social environment of people living with HIV/AIDS (PLWHA) in Thailand but have not been assessed in terms of their quality of life (QOL). In this study, we preliminarily examined the relationship between PLWHA's perception of social environment and QOL. The participants of this study were 200 PLWHA who belonged to the self-help groups in Chiang Mai province, northern Thailand. We collected data in face-to-face interviews using a structured questionnaire. The QOL was measured using a Thai version of the Medical Outcomes Study HIV Health Survey. Those who perceived themselves as well accepted by the community, perceived health services accessible or someone's help available, tended to have better QOL in terms of mental health. Community acceptance was most significantly related to QOL. Stratified analyses revealed similar relationships between perceived social environment and QOL among symptomatic and non-symptomatic participants but the relationships appeared weaker among men than women. In sum, our preliminary findings suggest that supportive social environment, especially community acceptance, is important for mental aspects of PLWHA's QOL irrespective of the disease stages and for women who usually take on multiple roles in the Thai traditional family. PMID- 16338771 TI - Late diagnosis of HIV infection among individuals with low, unrecognised or unacknowledged risks in England, Wales and Northern Ireland. AB - A small number of UK nationals who have a low, unrecognised or unacknowledged risk for HIV present late in the course of HIV infection; often after frequent attendances to primary care physicians. Information from in-depth interviews with individuals diagnosed with HIV in England, Wales and Northern Ireland (EW&NI) was analysed. Those diagnosed because of HIV-related symptoms (late diagnoses) were compared with those diagnosed for other reasons. Of the 286 individuals interviewed, 157 (55%) had HIV-related symptoms at the time of diagnosis, and 129 were tested for other reasons. A greater proportion of those diagnosed late were male and older. Of the 157 late diagnoses, 77 were considered to have acquired HIV heterosexually in the UK, 19 heterosexually abroad, 16 through 'high-risk' behaviours, 15 heterosexually by a 'high-risk' partner, four through blood transfusion and the remainder through an unusual or unknown route. A significantly higher proportion of those diagnosed late had had a long-standing relationship. None had been informed by a current or ex-partner of their HIV status. Primary care physicians should consider HIV as a possibility when patients without an apparent risk for HIV-infection present with symptoms indicative of possible immune suppression. Sensitive partner notification practices that enable a greater number of individuals to inform their partners should be explored. PMID- 16338772 TI - A qualitative study of persons who are 100% adherent to antiretroviral therapy. AB - This qualitative study examined the medication-taking behaviors and attitudes of participants determined to be 100% adherers to antiretroviral therapy from a NIH funded study testing a 12-week telephone adherence intervention. Using open-ended questions, interviewers collected data on a sample of 13 informants, whose medication adherence to a randomly selected antiretroviral medication was 100%, based on a 30-day data collection using electronic event monitoring (EEM). The analysis revealed 'successful medication management' as the core category or main theme. The participants achieved success with medication adherence through managing specific areas (regimen, self and environment). By adopting realistic expectations and pragmatic attitudes, adherence is fostered when medication taking is a priority, when patients believe in the efficacy of their medications and when there is a strong patient/provider relationship. Future research is needed to develop tailored interventions using strategies identified by this population. Further in-depth examination of medication-taking behaviors in 100% adherers may be useful in developing individualized programs to maximize adherence to antiretroviral therapy in the clinical setting. PMID- 16338773 TI - Strategies for management and treatment of dyslipidemia in HIV/AIDS. AB - With the improved survival of HIV-infected patients, there are increased concerns about the long-term effects of treatment, including protease inhibitor (PI) related dyslipidemia. Some 50-70% of patients receiving combination antiretroviral therapy (ART) involving PIs develop lipid abnormalities consisting of elevated levels of total cholesterol, low-density lipoprotein cholesterol and triglycerides that are well-known risk factors for cardiovascular disease. Treatment of HIV dyslipidemia should include lifestyle modifications such as a low-fat diet, increased exercise, reduced alcohol consumption and smoking cessation. In many patients, however, these changes alone will not correct lipid levels. In some patients, changing the PI component of ART to another PI or non PI and/or lipid-lowering drugs has proven successful. Each approach is associated with advantages and limitations and the need to maintain viral suppression must be balanced with the need to treat abnormal lipid levels. PMID- 16338774 TI - Risks for HIV infection among users and sellers of crack, powder cocaine and heroin in central Harlem: implications for interventions. AB - This article investigates behaviours that may be associated HIV infection among users and sellers of crack, powder cocaine and heroin in central Harlem. Chain referral sampling and other strategies were combined to acquire a sample of 637 (Users = 546; Sellers = 91) who provided urine specimens that were tested for the presence of drugs and HIV. Nearly a quarter (23.9%) of all respondents were HIV positive. Drug injectors were more than 2.5 times more likely to have HIV infections than other respondents (OR = 2.66; 95% CI 1.66-4.26). Those involved in frauds/cons were almost as likely to be HIV positive (OR = 2.58; 95% CI 1.64 4.06). Those with a marital status of being separated, divorced or widowed were twice as likely to be HIV infected (OR 2.16; 95% CI 1.43-3.25). Respondents currently having multiple partner sex (OR = 1.66; 95% CI 1.1-2.51) or who were female (OR = 1.66; 95% CI 1.12-2.45) were more than 1.5 times more likely to be HIV positive. Thus, controlling for lifetime drug injection and current multiple partner sex, other factors, such as participating in frauds/cons, as well as relationship status and being female, were also associated with HIV infection. PMID- 16338775 TI - HIV risk and communication between regular partners in a cohort of HIV-negative gay men. AB - This paper reports on the breaking of agreements between regular partners among HIV-negative gay men in Sydney. Data were from the 1333 men completing face-to face interviews through December 2003 for the Health in Men (HIM) open cohort of HIV-negative gay men in Sydney.822 men had a primary regular partner during the six month period before their 2003 interview. Most of these men had entered into agreements with their partners about sex either with each other or with other partners (87.2%). They most commonly agreed not to use condoms with each other (50.6%). Regarding casual sex, they most commonly agreed to always use condoms (34.2%) or to have no sex with men outside their relationships (28.6%). 48.8% reported some discomfort discussing with their partner their sex outside the relationship. Among those with agreements with their partners, 27.7% reported ever breaking those agreements. Those who found it more difficult to discuss issues of HIV serostatus and sexuality were more likely to report having broken their agreements (p<.001; p=.021 at one-year follow-up) and were more likely to have engaged in unprotected anal intercourse with casual partners (p<.001). A third of those men who broke their agreements did not inform their partner. A substantial proportion of gay men with agreements with their regular partners report some discomfort discussing sexuality and HIV serostatus with their partners. Difficulty discussing these issues may place these men at increased risk of breaking their agreements and may place both themselves and their partners at increased risk of infection. PMID- 16338777 TI - Microbiology profile in women with pelvic inflammatory disease in relation to IUD use. AB - OBJECTIVE: To study the microbial characteristics of patients with pelvic inflammatory disease (PID) and the possible impact of an intrauterine device (IUD) on the microbial environment in women presenting with PID. METHODS: Case control study, investigating 51 women with acute PID and 50 healthy women. Endocervical specimens for microbiological investigation were obtained at gynaecological examination. RESULTS: IUD users with PID had significantly more Fusobacteria spp. and Peptostreptococcus spp. than non-IUD users with PID. The finding of combinations of several anaerobic or aerobic microbes was associated with a significantly increased risk of PID and with complicated PID. In IUD users, the combinations of several anaerobic/aerobic microbes were associated with an increased risk of PID, irrespective of duration of IUD use. Long-term IUD use appeared to be associated with an increased risk of a PID being complicated. CONCLUSION: The finding of several anaerobic or aerobic microbes appears to be associated with PID in users of IUD. PMID- 16338778 TI - Pelvic inflammatory disease during the post-partum year. AB - OBJECTIVE: To investigate the occurrence of, and risk factors for, pelvic inflammatory disease (PID) occurring during the post-partum year. METHODS: Demographic and clinical data for women who delivered a term infant with 5-minute Apgar score > or = 8 from 1992 through 1999 at a large urban hospital were extracted from an electronic medical record system. RESULTS: During the study period, 15 206 deliveries occurred among 12 549 women. PID was diagnosed during the post-partum year of 148 (1.0%) deliveries. In univariate analysis, young age, black race, and both pre-delivery history and post-partum diagnosis of chlamydial and gonococcal infection were associated with PID. In multivariate analysis, only young age and a positive test for gonorrhea before delivery or post-partum were independent predictors of PID. CONCLUSIONS: Pelvic inflammatory disease was diagnosed during the post-partum year in 1% of women studied. Young maternal age was an important demographic risk factor. Further investigation of post-partum STD acquisition and progression to PID is needed to determine whether women are at increased risk following delivery. PMID- 16338779 TI - An evaluation of butoconazole nitrate 2% site release vaginal cream (Gynazole-1) compared to fluconazole 150 mg tablets (Diflucan) in the time to relief of symptoms in patients with vulvovaginal candidiasis. AB - BACKGROUND: It is estimated that as many as 13 million cases of vulvovaginal infection occur in the United States annually, the majority of which are the result of Candida albicans infection. The symptoms of vulvovaginal infections are often painful and distressing to the patient. The objective of this study was to compare the time to symptomatic relief of vulvovaginal candidiasis (VVC) with butoconazole nitrate 2% Site Release vaginal cream (Gynazole-1) and oral fluconazole 150 mg tablets (Diflucan). METHODS: This randomized, open-label, parallel study evaluated 181 female patients with moderate to severe symptoms of VVC. Patients were randomized to single-dose therapy with either butoconazole nitrate 2% Site Release vaginal cream or fluconazole. The primary outcome measure was the time to onset of first relief of symptoms. Secondary measures included the time to overall relief of symptoms and the reinfection rate over the first 30 days following treatment. The overall safety of both products was investigated through the collection of adverse event reports. RESULTS: The median time to first relief of symptoms occurred at 17.5 h for butoconazole patients as compared to 22.9 h for fluconazole patients (p < 0.001). The time at which 75% of patients experienced first relief of symptoms was 24.5 h versus 46.3 h for butoconazole and fluconazole, respectively (p < 0.001). By 12- and 24-h post-treatment, 44.4% and 72.8% of patients in the butoconazole treatment group reported first relief of symptoms versus 29.1% and 55.7% of patients in the fluconazole group (p = 0.044 and p = 0.024 respectively). In patients experiencing first relief of symptoms within 48 h of dosing, the median time to first relief of symptoms in the butoconazole treatment group was significantly shorter at 12.9 h compared to 20.7 h for the fluconazole treatment group (p = 0.048). There were no significant differences between the two groups with respect to time to total relief of symptoms or reoccurrence of infection within 30 days of treatment. Butoconazole therapy was shown to have fewer reported adverse events, including drug-related adverse events, than fluconazole therapy. Vulvovaginal pruritus and vulvovaginal burning were the most common drug-related adverse events attributed to butoconazole. Headache, diarrhea, nausea, upset stomach and skin sensitivity were the most common drug-related adverse events attributable to fluconazole. CONCLUSIONS: Single-dose butoconazole nitrate 2% Site Release vaginal cream provides statistically significant improvement in time to first relief of symptoms in the treatment of VVC compared to fluconazole. There is no difference between these two treatments with respect to total relief of symptoms or reinfection rate. Although there was no significant difference in the incidence of adverse events judged by the investigator to be treatment-related, butoconazole treatment did result in fewer patients experiencing adverse events than fluconazole. PMID- 16338780 TI - IL-1beta, IL-6 and IL-8 levels in gyneco-obstetric infections. AB - OBJECTIVE: During pregnancy cytokines and inflammatory mediators stimulate the expression of prostaglandin, the levels of which determine the onset of labor. The aim of this work was to study interleukin IL-1beta, IL-6 and IL-8 levels in the vaginal discharge, serum and urine of pregnant women with genitourinary infection before and after specific treatment. One hundred and fifty-one patients were studied during the second or third trimester of their pregnancy. METHODS: The selected patients were: healthy or control group (n = 52), those with bacterial vaginosis (n = 47), those with vaginitis (n = 37), those with asymptomatic urinary infection (n = 15) and post-treatment. The level of cytokines was assayed by ELISA test. The Mann-Whitney U-test was used for statistical analysis. RESULTS: The IL-1beta levels in vaginal discharge were: control 103.5 +/- 24.2 pg/ml, bacterial vaginosis 1030 +/- 59.5, vaginitis 749.14 +/- 66.7l ( p < 0.0001), post-treatment 101.4 +/- 28.7. IL-6 values were similar in both control and infected groups, and there were no patients with chorioamnionitis. In vaginal discharge IL-6: control 14.2 +/- 3.9 pg/ml, bacterial vaginosis 13.2 +/- 3.8, vaginitis 13 +/- 4.2. IL-8 levels were: control 1643 +/- 130.3 pg/ml, bacterial vaginosis 2612.7 +/- 257.7, vaginitis 3437 +/- 460 (p < 0.0001), post-treatment 1693 +/- 126.6. In urine the results were: control 40.2 +/- 17 pg/ml, asymptomatic urinary infection 1200.7 +/- 375 (p < 0.0001). In patients with therapeutic success both IL-1beta and IL-8 returned to normal levels. CONCLUSIONS: Genitourinary infections induce a significant increase in IL-1beta and IL-8 levels in vaginal secretions, and IL-8 in urine as well. Both cytokines could be useful as evolutive markers of infection. PMID- 16338781 TI - Periodontal disease and bacterial vaginosis increase the risk for adverse pregnancy outcome. AB - OBJECTIVES: To determine whether periodontal disease or bacterial vaginosis (BV) diagnosed before pregnancy increase the risk for adverse pregnancy outcome. METHODS: We enrolled a total of 252 women who had discontinued contraception in order to become pregnant. The first 130 pregnant women were included in the analyses. RESULTS: Multivariate analysis showed a strong association between periodontal disease and adverse pregnancy outcome (OR 5.5, 95% confidence interval 1.4-21.2; p = 0.014), and a borderline association between BV and adverse pregnancy outcome (OR 3.2, 95% confidence interval 0.9-10.7; p = 0.061). CONCLUSION: Our study suggests that pre-pregnancy counseling should include both oral and vaginal examinations to rule out periodontal disease and BV. This may ultimately have an impact on antenatal healthcare, and decrease the risk for adverse pregnancy outcome. PMID- 16338782 TI - Postpartum sterilization choices made by HIV-infected women. AB - OBJECTIVE: To assess if HIV-infected women made different choices for postpartum sterilization after implementation of the Pediatric AIDS Clinical Trials Group protocol 076 (November 1, 1994) compared to before implementation. STUDY DESIGN: A retrospective cohort study in which medical records were reviewed to obtain demographic, obstetric and HIV-related data from January 1993 through December 2002. HIV-infected women who completed a pregnancy by birth or abortion were divided into two comparison groups: "Pre-076" and "Post-076". The primary outcome was sterilization by postpartum tubal ligation.Results. Forty-two women (74%) in the Pre-076 group chose sterilization compared to 139 of 310 women (45%) in the Post-076 group (unadjusted OR 3.44, 95% CI 1.83, 6.47). Seventy-one percent of women younger than 21 years of age in the Pre-076 Group chose sterilization compared with only 35% of women younger than 21 years in the Post-076 group (p = 0.0136). Similarly, 78% of primiparous women chose sterilization after their first pregnancy in the Pre-076 group, compared to 14% in the Post-076 group (p < 0.001). CONCLUSIONS: Since the implementation of PACTG 076 protocol in November 1994, fewer HIV-infected women chose postpartum sterilization. The typical woman who now chooses postpartum sterilization is less likely to be young or primiparous than those who chose sterilization before PACTG Protocol 076 implementation. PMID- 16338783 TI - Transfer of meropenem in the ex vivo human placenta perfusion model. AB - OBJECTIVES: To determine maternal-fetal transplacental passage of meropenem in the ex vivo human perfusion model. STUDY DESIGN: Term placentae (n = 6) were collected immediately after delivery. A single cotyledon was localized, perfused and stabilized with physiologic Eagles minimal essential medium containing 3% bovine albumin and heparin as described by Chalier (Chalier JC. Criteria for evaluating perfusion experiments and presentation results. Contrib Gynecol Obstet 1985; 13:32 - 39). Meropenem was added to the maternal medium in concentrations similar to maternal serum peak and trough levels, then perfused through the maternal circulation of the cotyledon. To assess transfer and accumulation, fluid aliquots from both the maternal and fetal compartments were collected over an hour at defined intervals in an open and closed system. Antipyrine 14C was added to the medium in order to calculate the transport fraction and clearance indexes. Meropenem and antipyrine 14C concentrations were determined by High-pressure Liquid Chromatography and liquid scintillation, respectively. RESULTS: Mean antipyrine transport fraction was 2.33 + 0.25. Maternal and fetal mean meropenem peak concentrations were 54.3 + 3.3 microg/ml and 2.2 + 0.18 microg/ml, respectively. Whereas, maternal and fetal mean trough concentrations were 12.7 + 1.3 microg/ml and 0.41 + 0.10 microg/ml, respectively. Mean peak clearance index was 0.077 + 0.007 and the mean trough was 0.052 + 0.015. Mean accumulation for the peak and trough concentrations of meropenem were 0.9 and 2.95 microg/ml, respectively. CONCLUSIONS: Transplacental passage of meropenem was incomplete in the ex vivo human placental perfusion model. Accumulation was also noted in the fetal compartment. PMID- 16338784 TI - Malaria in pregnancy. AB - Recently, there has been a resurgence of malaria in densely populated areas of the United States secondary to human migration from endemic areas where factors such as cessation of vector control, vector resistance to insecticides, disease resistance to drugs, environmental changes, political instability, and indifference, have played a role for malaria becoming an overwhelming infection of these tropical underdeveloped countries. It is important for health care providers of gravida to be alert of the disease and its effects on pregnancy. PMID- 16338785 TI - Breast milk pasteurisation in developed countries to reduce HIV transmission. Do the benefits outweigh the risks? AB - BACKGROUND: Transmission of HIV through breastfeeding is well documented. The World Health Organisation advise HIV-infected women in developed countries to use alternatives to breastfeeding together with highly active antiretroviral therapy and optimal management of delivery to prevent transmission of HIV to their infant. CASE REPORT: We present the case of an HIV-infected woman electing to exclusively breastfeed for six months and applying milk pasteurisation techniques without transmission to her infant. Two paired samples of her breast milk were tested for HIV RNA prior to and after pasteurisation. The first pair of specimens reported no change in HIV RNA copy number, the second pair of specimens reported an increase in copy number. DISCUSSION: This technique, the evidence for HIV inactivation and the effects pasteurisation has on nutritional and immunological components of breast milk are discussed. CONCLUSION: In conclusion, we believe there is currently insufficient data to recommend this technique either as a safe alternative to formula feeding in resource-rich countries or as a method for providing intact immunological components of breast milk to the infant. PMID- 16338786 TI - Spontaneous midgestation abortion associated with Bacteroides fragilis: a case report. PMID- 16338787 TI - PKC isozymes and diacylglycerol-regulated proteins as effectors of growth factor receptors. AB - Growth factors exert their cellular effects through signal transduction pathways that are initiated by the ligation of growth factors to their cell surface receptors. One of the well-established effectors of growth factor receptors is protein kinase C (PKC), a family of serine-threonine kinases that have been known for years as the main target of the phorbol ester tumor promoters. While there is abundant information regarding downstream PKC effectors and partners, how individual PKC isozymes become activated by growth factors and the regulation of receptor function by PKCs is only partially understood. Moreover, the identification of novel "non-kinase" DAG-binding proteins has added a new level of complexity to the field of DAG signaling. PMID- 16338788 TI - c-Myb a stem-progenitor cell regulator in multiple tissue compartments. PMID- 16338789 TI - c-Yes response to growth factor activation. AB - Transmembrane receptors link the extracellular environment to the internal control elements of the cell. This signaling influences cell division, differentiation, survival, motility, adhesion, spreading and vesicular transport. Central to this signaling is the Src family of nonreceptor tyrosine kinases. The most studied kinase of this nine member family, c-Src, shares a similar structure, as well as a similar expression pattern to that of another Src family protein, c-Yes. Despite high conservation in sequence, molecular studies demonstrate that the functional domains of these kinases can contribute to specificity in signaling. At the cellular level, analysis of tight junction formation also serves as a model to differentiate c-Yes and c-Src signaling. Results suggest that c-Yes promotes formation of the tight junction by phosphorylating occludin, while c-Src signaling downregulates occludin formation in a Raf-1 dependent manner. In addition, pp62c-Yes knockout mice exhibit a specific physiological function phenotype that is distinct from c-src-/- mice. In these studies, c-yes-/- mice exhibit decreased transcytosis of pIgA from the blood to the bile, while c-src-/- mice exhibit deficits in osteoclasts function and bone resorption. Of particular interest in this review are receptor signals that specifically influence the actions of c-Yes. Growth factors that influence many Src family proteins include the PDGF-R, CSF-1 receptor and others. Since these receptors interact with various Src-family kinases, it is predicted that specific signaling is generated by differential recruitment to the cell membrane and/or differentiated interactions with substrates and binding partners. This review provides an overview of c-Yes interactions with specific receptor signaling pathways and how this interaction potentially influences the known physiological roles of c-Yes. PMID- 16338790 TI - Phe45 of NRG2beta is critical for the affinity of NRG2beta for ErbB4 and for potent stimulation of ErbB4 signaling by NRG2beta*. AB - The Neuregulins (NRGs) are members of the epidermal growth factor (EGF) family of growth factors. EGF family members regulate the signaling of ErbB family receptor tyrosine kinases, including the epidermal growth factor receptor (EGFR/ErbB1), ErbB2/HER2/Neu, ErbB3/HER3 and ErbB4/HER4. We have previously demonstrated that the EGF family hormone NRG2beta is a potent ErbB4 agonist, whereas NRG2alpha is a weak ErbB4 agonist. We have also previously demonstrated that Phe45 of NRG2beta regulates the potency of NRG2beta. Here, we address the hypotheses that Phe45 regulates the potency of NRG2beta by regulating the affinity of NRG2beta for ErbB4. We demonstrate that Phe45 of NRG2beta indeed regulates the affinity of NRG2beta for ErbB4. Furthermore, a hydrophobic or uncharged amino acid side chain at residue 45 contributes to NRG2beta binding to ErbB4. These data indicate that Phe45 of NRG2beta may regulate the affinity of NRG2beta for ErbB4 by interacting with hydrophobic amino acids in ErbB4. PMID- 16338791 TI - Effect of epiregulin on pancreatic beta cell growth and insulin secretion. AB - The aim of this study was to determine whether epiregulin, a novel member of EGF related growth factor family, was able to affect proliferation and secretory function of rat insulinoma INS-1E and RINm5F cell lines. A 24 h treatment with epiregulin resulted in a stimulation of INS-1E and RINm5F cells proliferation; this effect was completely blocked in the presence of an anti-epiregulin antibody which did not affect basal DNA synthesis in the absence of added ligand. In acute experiments, epiregulin was able to potentiate insulin release in the presence of glucose or arginine, in the two cell lines. Finally, in the two cell lines expressing ErbB receptors, we demonstrated that only EGFR/ErbB1 was activated by epiregulin. Thus, epiregulin appears as a new growth and insulinotropic factor in pancreatic beta cell lines. PMID- 16338792 TI - Analysis of fibroblast growth factor 2 (FGF2) gene transcription and protein distribution in the bovine testis. AB - Several fibroblast growth factors (FGFs) are implicated in proliferation and differentiation of both somatic and germ cells during testicular development, as well as in spermatogenesis of adult testis. The expression of FGF2 was studied in the adult bovine testis using quantitative RT-PCR, RNA in situ hybridization, and immunohistochemistry. Quantitative RT-PCR revealed consistent levels of FGF2 mRNA in parenchymal samples of the bovine testis. In situ hybridization localized FGF2 transcripts only in a constant fraction of Leydig and Sertoli cells as well as in modified Sertoli cells of the terminal segments. Immunohistochemistry revealed (a) no FGF2 protein in Sertoli cells (b) moderate cytoplasmic staining in Leydig cells and spermatogonia and (c) strong nuclear and faint cytoplasmic staining in myofibroblasts, in epithelial cells of straight tubules and rete testis and in blood vessels. These observations indicate a pleiotropic effect of FGF2 on the control of spermatogenesis in a paracrine and/or autocrine manner. PMID- 16338793 TI - Neurotrophin system expression in human pulmonary carcinoid tumors. AB - Neurotrophin (NT) and NT receptor expression was assessed in 12 typical (TC) and 8 atypical (AC) human pulmonary carcinoids by Western blot and immunohistochemistry. TC and AC carcinoid express to different extent NT and NT receptor proteins. Nerve growth factor (NGF) was expressed by 83% of the TC but not by the AC carcinoids. Brain derived neurotrophic factor (BDNF) was expressed by 33 and 100% of TC and AC carcinoids, respectively. NT-3 was expressed by 58% of the TC and 38% of AC carcinoids. TC carcinoids express high affinity NT receptors while 50% of the AC carcinoids express the TrkB receptor. Our results demonstrate that NGF/TrkA and BDNF/TrkB signaling need to be considered as regulatory pathways that may address survival, differentiation and/or aggressiveness of human pulmonary carcinoids. Contrarily to the BDNF/TrkB, expression of the NGF/TrkA signaling may overcome aggressiveness of carcinoid cells. NTs may be useful as markers in the clinic. PMID- 16338794 TI - Combined effects of TGFbeta1 and BMP2 in serum-free chondrogenic differentiation of mesenchymal stem cells induced hyaline-like cartilage formation. AB - This study investigated the effects of TGFbeta1, BMP2 or a combination of both on the chondrogenic differentiation of mesenchymal stem cells (MSCs) in a serum-free micromass culture system in vitro. Putative MSCs harvested from the iliac crest of 4-5 month old New Zealand White Rabbits were expanded and cultured in three dimensional high density micromass aggregate cultures containing TGFbeta1, BMP2 or a combination of both, in the absence of serum. After 14-20 days of culture, chondrogenic differentiation of the MSCs was assayed by toluidine blue staining, immunohistochemistry and semi-quantitative RT-RCR of type I collagen (CI) and type II collagen (CII). Quantitative measurements of cell proliferation and sulfated glycosaminoglycan (s-GAG) were also carried out to assess the growth rate and matrix deposition of the cultured aggregates. Both immunohistochemical staining and semi-quantitative RT-PCR showed that the combination of BMP2 and TGFbeta1 resulted in a marked enhancement of collagen II synthesis, with minimal collagen I expression, which would suggest hyaline-like cartilage formation. Additionally, BMP2 and TGFbeta1 had a synergistic effect on matrix proteoglycan deposition, as assessed by metachromatic toluidine blue staining. This is consistent with the quantitative measurement of glycosaminoglycans, whereby a significant increase in GAG/DNA was noted in the co-treatment group. Hence, it can be concluded that the combination of BMP2 and TGFbeta1 has a synergistic effect on the differentiation of MSC into hyaline-like cartilage tissue. PMID- 16338796 TI - What gets missed when deploying new technologies in A&E? AB - This paper presents a longitudinal study (over 4 months) of an A&E department where the existing whiteboards were replaced with PC-based computer systems. The study was conducted in two parts - an observation of the physical whiteboard usage and in-depth interviews with all users of both the traditional whiteboard usage and the replacement technology. The research was conducted with the systems manager and all whiteboard users (i.e. nursing management, nurses, doctors, porters, and agency staff) across a spread of time frames. Although the technology supported simple information requirements, complex co-ordination, collaboration, and awareness issues were left unsupported. The important role of a pen-holder (information co-ordinator) was poorly supported by the replacement technology as was the task of annotating information with changing situations and needs. Specific deployment issues are derived from these findings that should guide designers when implementing technology replacements for current physical information formats (e.g. whiteboards, notice boards, shared paper notes). PMID- 16338797 TI - The barriers to clinical coding in general practice: a literature review. AB - Clinical coding is variable in UK general practice. The reasons for this remain undefined. This review explains why there are no readily available alternatives to recording structured clinical data and reviews the barriers to recording structured clinical data. Methods used included a literature review of bibliographic databases, university health informatics departments, and national and international medical informatics associations. The results show that the current state of development of computers and data processing means there is no practical alternative to coding data. The identified barriers to clinical coding are: the limitations of the coding systems and terminologies and the skill gap in their use; recording structured data in the consultation takes time and is distracting; the level of motivation of primary care professionals; and the priority within the organization. A taxonomy is proposed to describe the barriers to clinical coding. This can be used to identify barriers to coding and facilitate the development of strategies to overcome them. PMID- 16338798 TI - Proving the concept of a data broker as an emergent alternative to supra enterprise EPR systems. AB - Electronic Patient Records systems configured into large enterprise models have become the assumed best route forward. In England, as in several other countries, this has expanded to a major meta-enterprise procurement programme. However, concerns are raised that such systems lack user ownership, and experience from other sectors shows difficulties with large enterprise systems. At a time of great change and once again shifting organizations, is this move simply building large and ponderous edifices with unstable materials? Latest software engineering research is now demonstrating the potential of an alternative model, enabling trusted information brokers to search out in real time at point of use data held in registered local and departmental systems. If successful, this could enable a new and less cumbersome paradigm. The data could move where needed whatever the service configuration. A concept demonstrator has been built set in the context of health and social care in England. It is important for all technological support to the health sector to be reviewed as new technologies emerge so as to identify and exploit new opportunities, and the results of this 3 year project show that the health record information broker route merits further investigative research. PMID- 16338799 TI - Implementing digital resources for clinicians' and patients' varying needs. AB - This paper presents an overview of several evidence-based medicine and patient information studies conducted across the UK health service over a 4 year period, investigating clinicians', managers', and patients' perceptions of digital resources (primarily digital libraries) in hospitals, Primary Care Trusts, NHS Direct (patient call centre) and patient groups. In-depth interviews and focus groups are analysed using grounded theory methodologies and through content analysis used to produce quantitative finding. The perceived impacts of three different methods employed for delivering health informatics are presented. The findings highlight some generic issues relevant for health informatics in the UK health sector as well as some specific issues for medical digital libraries. This paper reviews in more detail the issues of medical technology implementation (traditional implementation, on the wards, and intermediaries within in communities). A breakdown of the clinicians' and patients' information journey (information initiation, facilitation and interpretation) is also presented with regard to medical digital libraries and online resources. Broad guidelines derived from these findings are provided for health-informatics deployment. PMID- 16338800 TI - IT-driven patient safety at the heart of radiology service improvement. AB - Patient safety is a fundamental requirement of modern health-care systems and the application of information technology (IT) to this activity should have improvements in the area as one of its goals. Indeed, ensuring that the diagnostic IT strategy is optimized, for example, the use of IT in service redesign or data analysis, forms one of the main platforms for the National Framework for Service Improvement in Radiology. This paper presents both the concept behind and the results of a project that has been under way in the UK involving St Helens and Knowsley NHS Trust and IRS Ltd concerned with implementing effective IT-driven scientific support in the field of medical radiation protection. Locally developed software is employed in assessing, managing, and analysing patient dose data arising from X-ray examinations performed in the busy department of a large district general hospital (DGH). Such data are analysed in a variety of ways, for example, over time and according to location (department or X-ray room). This analysis not only provides a measure of performance against nationally agreed dose reference levels (DRLs) but also enables a detailed analysis of any variations as well as the establishment of local DRLs (performance indicators). This process provides quantitative input to management strategies aimed at service improvement. Such strategies are geared towards the support of the Ionizing Radiations (Medical Exposures) Regulations 2000 as well as implementation of quality-management principles at the heart of radiology practices. PMID- 16338801 TI - Temporal and spatial organization of doctors' computer usage in a UK hospital department. AB - This paper describes the use of an application accessible via distributed desktop computing and wireless mobile devices in a specialist department of a UK acute hospital. Data (application logs, in-depth interviews, and ethnographic observation) were simultaneously collected to study doctors' work via this application, when and where they accessed different areas of it, and from what computing devices. These show that the application is widely used, but in significantly different ways over time and space. For example, physicians and surgeons differ in how they use the application and in their choice of mobile or desktop computing. Consultants and junior doctors in the same teams also seem to access different sources of patient information, at different times, and from different locations. Mobile technology was used almost exclusively during the morning by groups of clinicians, predominantly for ward rounds. PMID- 16338802 TI - Representing different views of acupuncture in a single ontology. AB - A major challenge for health informatics is to model the health-care domain knowledge into appropriate and useful ontologies. This is difficult, and modellers tend to simplify things dramatically by ignoring the fact that the health care domain encompasses a global perspective. The influx of complementary and alternative medicines from the East to the West, or indeed the ongoing colonialization of orthodox Western medicine into other non-western traditions requires multiple paradigms of delivering treatment to be considered simultaneously, side by side. Models and specifications need to be developed to encompass this richness and represent this complex reality so as to understand what the different concepts and terms in each tradition achieve. Acupuncture is used as a test case of cross-membership between two different paradigms. Here, we briefly outline requirements and answer possible objections to our approach before illustrating how we have modelled heterogeneous domain knowledge from different cultures, and across different paradigms, within a single ontology. PMID- 16338803 TI - Measuring clinical performance using routinely collected clinical data. AB - Following the well-publicized problems with paediatric cardiac surgery at the Bristol Royal Infirmary, there is wide public interest in measures of hospital performance. The Kennedy report on the BRI events suggested that such measures should be meaningful to the public, case-mix-adjusted, and based on data collected as part of routine clinical care. We have found that it is possible to predict in-hospital mortality (a measure readily understood by the public) using simple routine data-age, mode of admission, sex, and routine blood test results. The clinical data items can be obtained at a single venesection, are commonly collected in the routine care of patients, are already stored on hospital core IT systems, and so place no extra burden on the clinical staff providing care. Such risk models could provide a metric for use in evidence-based clinical performance management. National application is logistically feasible. PMID- 16338804 TI - The Tees Confidentiality Model: mechanisms for implementing the sealed envelope. AB - This paper offers mechanisms capable of implementing the authorization functionality to be supported by the NHS Care Records Service. The patient confidentiality model for the Care Records Service includes restricting access to data by placing the data in a Sealed Envelope; providing access to data based on Legitimate Relationship, and other concepts; and the overriding of access restrictions in extraordinary or emergency situations. We informally show through examples how the Tees Confidentiality Model, a sophisticated model of authorization, can be used to implement Care Records Service authorization functionality to the level currently proposed, and also to much greater levels if they ever were to be required. The mechanisms discussed include using a range of permission types, called Confidentiality Permission Types; processing Confidentiality Permissions in a defined order according to complexity of type; using negative permissions to deny access; and providing override mechanisms for negative permissions. PMID- 16338805 TI - The ISABEL user survey. AB - ISABEL is a web-based clinical decision-support system for use by health care professionals. The Web site has been developed by the ISABEL Medical Charity. The system has come to the attention of the Department of Health, which is examining its potential effectiveness in the wider clinical context and exploring options for promoting its wider use in the NHS. The objectives of the work reported here were to review the existing use of ISABEL and to identify impediments to its development. A questionnaire was sent by e-mail to selected users of the system. Based on an analysis of the results (n=518), we found ISABEL to be a useful tool with many users. We believe that there is evidence of its success sufficient to support its continued availability and development. However, the largest hurdles to its increased use are systemic ones within the NHS and the way services are delivered. PMID- 16338806 TI - Usability: a critical success factor for managing change in the clinical info structure. AB - There can be no doubt that the clinical info-structure is being significantly enriched with the deployment of new systems throughout the health sector. From a technological perspective, the initial emphasis has been mainly on functionality and only latterly on the usability of these clinical information systems. However, the large scale and rapid pace of the changes being wrought in the health sector will have a major impact on clinicians and patients, not least in how they interact with the technology. Therefore, it is not only hardware and software but people-ware, too, that needs to be actively managed; not simply a one-off functional specification but an ongoing, complex relationship. Usability is the human factor that encompasses the ethical, educational, and evaluative aspects of design. There is also a strong case for regarding usability of clinical information systems as a key critical success factor for the management of change within the health-care domain. In particular, the relationship between usability, and education and training is examined. PMID- 16338807 TI - Religion, spirituality and aging. PMID- 16338808 TI - Is there a process of spiritual change or development associated with ageing? A critical review of research. AB - This review considers whether research shows a process of spiritual change or development associated with ageing. Spirituality was understood as that which is central to a sense of meaning and purpose in an individual's life and pertains to the sacred or transcendent. Electronic literature searches were conducted to find research published 1985-2003 aimed at understanding spiritual change, themes and tasks in later life. A total of 13 studies were reviewed that looked at changes in spirituality over time, spiritual themes and tasks in a lifespan development context and Tornstam's (Torstam, L. (1996). Gerotranscendence--a theory about maturing into old age. Journal of Aging & Identity, 1, 37-50) theory of gerotranscendence. The research reviewed suggested that some aspects of spirituality remain stable into old age but that there are identifiable spiritual tasks, needs and changes associated with ageing. Some common spiritual themes identified across the research were integrity, humanistic concern, changing relationships with others and concern for younger generations, relationship with a transcendent being or power, self transcendence, and coming to terms with death. These were not related to age per se, but to some of the challenges that age presents, and were mediated by cultural factors and individual differences. The findings and their limitations were discussed. PMID- 16338809 TI - Higher church attendance predicts lower fear of falling in older Mexican Americans. AB - Several studies have shown that involvement in religious activity appears to benefit health. To estimate the association between church attendance and fear of falling, we used a sample of 1341 non-institutionalized Mexican-Americans aged 70 and over from the third wave (1998-1999) of the Hispanic Established Population for the Epidemiological Study of the Elderly, followed until 2000-2001. Baseline potential predictors of fear of falling were church attendance, socio demographics, history of falls, summary measure of lower body performance (tandem balance, eight-foot walk, and repeated chair stands), functional status, depressive symptoms, cognitive status, and medical conditions. Fear of falling at the two-year follow-up was measured as no fear, somewhat afraid, fairly afraid, and very afraid. Chi-square statistic and multiple logistic regression analysis were used to estimate associations between the outcome and the potential predictors. Multiple logistic regression analysis showed that frequent church attendance was an independent predictor of lower fear of falling (odds ratio = 0.73, 95% confidence interval 0.58-0.92, P = 0.008) two years later. Other independent predictors of fear of falling were female gender, poorer objective lower body performance, history of falls, arthritis, hypertension, and urinary incontinence. Frequent church attendance is associated with decreased fear of falling in older Mexican-Americans. PMID- 16338810 TI - Relationship between adherence to tradition and depression in Chinese elders in China. AB - This study examined the relationship between adherence to tradition and depression in a large sample (N = 1502) of Chinese elders living in the People's Republic of China. In Chinese traditional culture, an elder's purpose, meaning and self-worth are derived, in large, from their social roles within the family and community. As the traditional culture declines so do these familiar roles, supports, and ways of coping. This constitutes a major loss which can have a variety of psychosocial consequences, one of which could be depression. The elders in our study responded to a nine-item measure of traditional mutual aid and intergenerational exchange. Depression was measured by the Center for Epidemiological Studies Depression Scale. Regression analysis found a significant and negative relationship between tradition and depression which suggests that adherence to tradition may have protective benefits. These findings have application with elders in China as well as with those who are immigrants to other societies. PMID- 16338811 TI - Neuropsychological functioning and delusions in dementia: a pilot study. AB - The present pilot study investigated the pattern of neuropsychological functioning associated with the presence of delusions in mild-to-moderate dementia. Participants, all of whom met criteria for dementia, were divided into two groups, delusional (n = 9) and non-delusional (n = 9). Individuals with hallucinations were excluded. Participants completed a neuropsychological test battery. Global cognitive functioning (MMSE) and behavioral disturbance (BEHAVE AD) were also assessed. Differences between the delusional and non-delusional group were most marked for immediate recall of stories, which was higher in the non-delusional group. Scores on semantic fluency, attention (mental control), and overall cognitive functioning (MMSE) were also lower in the delusional group. Conversely, simple attention span (Digit Span) was within normal limits in both groups. Floor effects were noted on measures of delayed recall and alternating attention. This study supports previous findings of greater neuropsychological impairment in delusional as compared to non-delusional individuals with dementia. However, some areas of cognitive functioning may be relatively preserved. Future research should examine semantic processing in persons with dementia with and without delusions. PMID- 16338812 TI - Use of the Minimum Data Set--Home Care: a cluster randomized controlled trial among the Chinese older adults. AB - The objective of this study was to test whether the utilization of Minimum Data Set-Home Care (MDS-HC) as a case finding instrument for Chinese attenders of elderly health centres in Hong Kong had a beneficial impact on the physical and mental health status of those older clients. The dependent variables were 13 outcome variables measuring different dimensions of participants' physical and mental health status. Using a cluster randomized controlled trial design, we assigned three of six participating elderly health centres to the experimental group and the other three to the control group. The respondents were 925 (472 in the experimental group and 453 in the control group) elderly health centre attenders aged 65 years or above. They were assessed by MDS-HC and 734 (382 in the experimental group and 352 in the control group) of them were re-assessed one year later. In the experimental group, the medical doctors had been given their patients' profile of potential problem areas identified by MDS-HC before they treated their patients whereas in the control, the medical doctors treated their patients with their usual practice. We found that older attenders in the experimental group improved more than the ones in the control group in only two out of 13 areas. Surprisingly, the patients in the experimental group deteriorated even more than the ones in the control group in terms of bowel incontinence. Our results indicated that the function of the MDS-HC in case finding or screening is limited in the Hong Kong primary medical care setting. PMID- 16338813 TI - Comparative validity of seven scoring systems for the instrumental activities of daily living scale in rural elders. AB - Lawton and Brody's eight-item Instrumental Activities of Daily Living (IADL) scale is used often with elderly patients but scored in several different ways. We scored the IADL with seven popular procedures ranging from relatively simple to complex (Guttman scores, summed and Rasch scores from dichotomous, trichotomous, and polytomous items) in a sample of rural elders (N = 231). We compared the IADL scales' prediction of concurrent cognitive functioning, depressive symptoms, psychosocial functioning, and health care use (medications, outpatient visits, inpatient days). Validity coefficients ranged from small to large among outcome variables but were highly consistent across IADL scoring procedures. Consequently, researchers and clinicians may prefer to use simpler IADL scoring procedures with this population. PMID- 16338814 TI - Principles and practice of geriatric assent. AB - Geriatric assent involves health care professionals' active collaboration with cognitively impaired patients that takes account of their longstanding values in any major health care decisions. The main purpose of this paper is to assist geriatric health practitioners 'in the field' to understand how to apply geriatric assent in a variety of clinical situations to maximize incapacitated older adults' input into decision-making. A case example and algorithm are presented to illustrate the basic principles of implementing geriatric assent. Practice informed by the principles of geriatric assent will preserve respect for the current and future autonomy of patients across diverse cultural backgrounds. PMID- 16338815 TI - 'It's like a revolving door syndrome': professional perspectives on models of access to services for people with early-stage dementia. AB - As the need for services for people with dementia grows, and the benefits of early intervention become clear, it has become important to understand what factors may improve access to services for people with early-stage dementia. There are a number of models of service access, and these emphasise different areas, whether individual factors, relationships, or social context. The relevance of these models to variations in service access for people with early stage dementia, and how well they relate to professional accounts, is not well known. In this study, 30 key professionals were interviewed about access to services for people with early-stage dementia in order to explore how different models of access were reflected in their own understandings. People with early stage dementia were thought to have a range of complex needs, but participants felt these needs remained largely unmet. When articulating the reasons why they thought needs were unmet, participants focused on the impact of the framework within which services are delivered. The findings highlight the importance of considering relationships and socio-contextual factors, such as the impact of the framework of service delivery, when attempting to understand variations in access to services. In order to improve access to services, it will be necessary to move beyond addressing individual factors relating to access, and to consider the impact of the framework for service delivery and the relationships that influence contact with services. PMID- 16338816 TI - Interpersonal dependency in older adults and the risks of developing mood and mobility problems when receiving care at home. AB - Despite a broad interest in various types of dependency as they relate to older people (structured dependency, learned dependency, learned helplessness, and interdependency), research of dependency in older people has not included an individual difference measure of interpersonal dependency. Studies that have examined the correlates of interpersonal dependency in general populations have found links with mental health conditions such as depression and anxiety and also with physical illness. If these findings could generalise to older populations then there would be important intervention and financial implications for providers of health services to older adults. This research examined the correlates of interpersonal dependency in older adults using a new measure of interpersonal dependency-the Interpersonal Dependency Scale for Older Adults (IDS OA). One hundred and five new clients aged 65-90 years recruited through a private home care agency responded to a postal survey requesting their completion of a questionnaire package. Interpersonal dependency was found to correlate significantly with both depression and mobility. In addition, a hierarchical regression analysis found that both higher interpersonal dependency and depression were significant positive predictors of poor mobility in older adults. PMID- 16338817 TI - Cross-cultural study comparing the association of familism with burden and depressive symptoms in two samples of Hispanic dementia caregivers. AB - Familism has been pointed out as a key value in Hispanic culture that may or may not be associated with caregiver distress. Although groups included in the Hispanic identity have many common features, differences between Hispanic sub groups in the relationship of familism with burden and depressive symptoms remain unexplored. The association of familism with burden and depressive symptomatology was examined in 48 Hispanic dementia caregivers from Los Angeles (CA, USA) and 60 from Madrid (Spain) through path analyses. Burden and depressive symptomatology were positively and significantly related in both samples. Familism was significantly correlated with lesser burden in the USA Hispanic caregiver sample, but with higher levels of depressive symptoms in the Spanish sample. Significant differences between Hispanic samples were found in the relationship between familism, burden, and depression, denoting the importance of specific cultural contexts influencing dementia caregiving in Hispanics. PMID- 16338819 TI - Comparison of bilateral whisker movement in freely exploring and head-fixed adult rats. AB - Rats move their whiskers actively during tactile exploration of their environment. The whiskers emanate from densely innervated whisker follicles that are moved individually by intrinsic facial muscles and as a group by extrinsic muscles. Several descriptions of whisker movements in normal adult rats during unrestrained exploration indicate that rats move their whiskers in the 6-9 Hz range when exploring a new environment. The rate can be elevated to nearly 20 Hz for brief episodes just prior to making a behavioural decision. The present studies were undertaken to compare whisker dynamics in head-restrained and freely moving rats with symmetrical or asymmetrical numbers of whiskers on the two sides of their face and to provide a description of differences in whisker use in exploring rats after trimming all but two whiskers on one side of the face, a condition that has been shown to induce robust cortical plasticity. Head-fixed rats were trained to protract their whiskers against a contact detector with sufficient force to trigger a chocolate milk reward. Whisker movements were analyzed, and the results from head-fixed animals were compared with free-running animals using trials taken during their initial exploration of novel objects that blocked the rat's progress down an elevated runway. The results show that symmetrical whisker movements are modulated both by the nature of the task and the number of whiskers available for exploration. Rats can change their whisker movements when the sensitivity (threshold) of a contact detector is raised or lowered, or when the nature of the task requires bilateral input from the whiskers. We show that trimming some, but not all whiskers on one side of the face modifies the synchrony of whisker movement compared to untrimmed or symmetrically trimmed whiskers. PMID- 16338820 TI - Alteration of sensorineural circuits in spinal cord by chronic contact dermatitis. AB - In the present study, eczema-induced alteration of sensorineural circuits of the spinal dorsal horn was investigated. Eczematous lesions resembling atopic dermatitis were induced by repeated application of diphenylcyclopropenone (DCP) onto murine right hind paws. Immunohistochemical labeling of calcitonin gene related peptide and substance P was increased in the dorsal horn on the DCP treated side. Expression of calcium binding proteins, calretinin and calbindin D28K, normally widely seen in dorsal horn interneurons, was up-regulated on the DCP-treated side. E-Cadherin and alpha-N-catenin, synapse-related molecules, were intensely expressed in the spinal dorsal horn of the DCP-treated side. Interestingly, c-Fos positive cells were also significantly increased in laminae I and III of the DCP-treated side. These results suggest an enhanced release of neuropeptides from peripheral afferents and alterations in the sensorineural circuitry of the dorsal horn. These changes may account for the enhanced sensory sensitivity recognized in patients with chronic eczema and atopic dermatitis. PMID- 16338821 TI - Stanley "Sandy" J. Bolanowski (22 February 1950-13 January 2005). PMID- 16338822 TI - Perception of the tactile texture of raised-dot patterns: a multidimensional analysis. AB - An ALSCAL multidimensional scaling analysis in Euclidean space revealed that three orthogonal perceptual dimensions can account for the judged tactile dissimilarities of raised-dot patterns. Through magnitude estimates of various perceptual attributes, it was determined that the three dimensions consist of blur, roughness, and clarity. The only effect that selective adaptation of the Pacinian (P) channel had was to change the perceptual clarity of the raised dots against their background. Adaptation of the P channel with a 20 dB SL 250 Hz stimulus enhanced clarity. As indicated by magnitude estimates, adaptation of the P channel by the 250 Hz stimulus had no effect on the perceived roughness of the dot pattern but did cause the individual dots of the textured pattern to feel smoother. When the observer was required to estimate magnitude "overall roughness" defined as a combination of dot-pattern roughness and individual-dot roughness, adaptation of the P channel affected perceived roughness by reducing it. Taken as a whole, the results are consistent with the hypothesis that the NP channels and the P channel jointly influence the perception of textured surfaces. PMID- 16338823 TI - Genetic analysis of barrel field size in the first somatosensory area (SI) in inbred and recombinant inbred strains of mice. AB - We measured the combined area of posterior medial barrel subfield (PMBSF) and anterior lateral barrel subfield (ALBSF) areas in four common inbred strains (C3H/HeJ, A /J, C57BL /6J, DBA/2J), B6D2F1, and ten recombinant inbred (RI) strains generated from C57BL/6J and DBA/2J progenitors (BXD) as an initial attempt to examine the genetic influences underlying natural variation in barrel field size in adult mice. These two subfields are associated with the representation of the whisker pad and sinus hairs on the contralateral face. Using cytochrome oxidase labeling to visualize the barrel field, we measured the size of the combined subfields in each mouse strain. We also measured body weight and brain weight in each strain. We report that DBA/2J mice have a larger combined PMBSF/ALBSF area (6.15 +/- 0.10 mm(2), n = 7) than C57BL /6J (5.48 +/- 0.13 mm(2), n = 10), C3H/HeJ (5.37 +/- 0.16 mm(2), n = 10), and A/J mice (5.04 +/ 0.09 mm(2), n = 15), despite the fact that DBA/2J mice have smaller average brain and body sizes. This finding may reflect dissociation between systems that control brain size with those that regulate barrel field area. In addition, BXD strains (average n = 4) and parental strains showed considerable and continuous variation in PMBSF/ALBSF area, suggesting that this trait is polygenic. Furthermore, brain, body, and cortex weights have heritable differences between inbred strains and among BXD strains. PMBSF/ALBSF pattern appears similar among inbred and BXD strains, suggesting that somatosensory patterning reflects a common plan of organization. This data is an important first step in the quantitative genetic analysis of the parcellation of neocortex into diverse cytoarchitectonic zones that vary widely within and between species, and in identifying the genetic factors underlying barrel field size using quantitative trait locus (QTL) analyses. PMID- 16338824 TI - Effects of high-frequency skin stimulation on SI cortex: mechanisms and functional implications. AB - Optical intrinsic signal (OIS) imaging methods were used to record the responses of contralateral SI cortex to 25 Hz ("flutter") and also to 200 Hz ("vibration") stimulation of the skin. Anesthetized cats and squirrel monkeys were subjects. Separate series of experiments were carried out to evaluate the contralateral SI response to continuous, multisecond 25 Hz vs. 200 Hz stimulation (a) at multiple skin sites arranged along the proximal-distal axis of the fore- or hindlimb (Series I); (b) in the presence and absence of a ring placed in firm contact with the skin surrounding the stimulus site (Series II); (c) before and after topical application of local anesthetic to the stimulus site (Series III); and, finally, (c) to continuous 25 Hz or 200 Hz stimulation applied independently, and also concomitantly ("complex waveform stimulation") to the same skin site (Series IV). The principal findings are: (a) the relationship between the SI optical responses to 25 Hz vs. 200 Hz stimulation of a skin site varies systematically with position of the stimulus site on the limb-at a distal site both 25 Hz and 200 Hz stimulation evoke a well-maintained increase in absorbance, and as the stimulus site is shifted proximally on the limb the response to 200 Hz, but not the response to 25 Hz stimulation, converts to a frank decrease in absorbance; (b) placement of a ring about a skin site at which in the absence of a ring 200 Hz stimulation evoked a decrease in SI absorbance converts the response to 200 Hz to one consistent with increased SI RA neuronal activation (i.e., with the ring in place 200 Hz stimulation evokes a change in SI absorbance approximating the response to 25 Hz stimulation); (c) topical local anesthetic preferentially and reversibly decreases the magnitude of the absorbance increase associated with 25 Hz flutter stimulation; and (d) complex waveform stimulation consistently is associated with a smaller increase in absorbance than obtained with same-site 25 Hz stimulation. Collectively, the findings are consistent with the idea that the Pacinian (PC) afferent activity which unavoidably accompanies cutaneous flutter stimulation triggers CNS mechanisms that "funnel" (sharpen) the spatially distributed contralateral SI response to the flutter stimulus. Viewed in this context, the fact that a flutter stimulus unavoidably co-activates RA and PC afferents appears functionally beneficial because the CNS mechanisms activated by PC afferent drive modify the SI response to skin flutter in a manner predicted to enable more accurate perceptual localization than would be possible if the flutter stimulus only activated RA afferents. PMID- 16338825 TI - Cortical control for mastication in cats: changes in masticatory movements following lesions in the masticatory cortex. AB - In a previous paper (Hiraba and Sato 2004) we reported that an accurate mastication might be executed by the cortical processing in bilateral masticatory area (MA)and motor cortices. The aim of this study was to determine if cats with lesion of either unilateral or bilateral MA showed changes in mastication. After exploring mechanoreceptive fields and motor effects of mastication-related neurons (MRNs) in MA using the single unit recording and intracortical microstimulation methods, we made various lesions in MAs with injections of kainic acid (0.1%, 2.0 microl). Since the MA was divided into facial (F) and intraoral (I) projection areas as reported in the previous paper, cats with the unilateral lesion in F or I, and with the bilateral lesion in F and F, I and I or F and I (F on one side and I on other side) were prepared. Cats with unilateral lesion in F or I and with bilateral lesion in F and I showed no changes in mastication except for prolongation of the food intake and masticatory periods. Cats with bilateral lesion into F and F, or I and I showed wider jaw-opening during mastication. Particularly, the latter group showed enormous jaw-opening, delay in the start of mastication and difficulty in manipulating food on the tongue. In all cats with lesions of each type, masticatory and swallowing rhythms remained normal. These findings suggest that accurate mastication is executed by the close integration between F and F and I and I of the bilateral MA. PMID- 16338826 TI - Cortical control of mastication in cats. 2. Deficits of masticatory movements following a lesion in the motor cortex. AB - Our previous study suggested that area P in the lateral wall of the presylvian sulcus and MA (masticatory cortex) in the rostral part of the orbital gyrus play an important role in execution of mastication. The aim of this present study is to examine if changes in orofacial behaviors and masticatory movements occur in cats with lesions of area P. First, we explored the locations in area P through the use of single unit recording and ICMS (intracortical microstimulation). Since mastication related neurons (MRNs) with the mechanical receptive field (RF) in facial or intraoral region were intermingled in area P, we performed either a partial or entire lesion in area P by injections of 2 microl or 4 microl of 0.1% kainic acid. Cats with the entire lesion in area P showed a decrease of food intake rates associated with abnormal tongue protrusion and wide jaw-opening, fluctuation of masticatory start, and prolongation of masticatory and food intake periods. Abnormal movements of tongue and jaw did not recover, although their prolongation and fluctuation returned to normal levels in one month. On the other hand, all deficits evoked by cats with the partial lesion recovered in about one month. In cats with the partial and entire lesions, masticatory rhythm remained normal. These findings suggest that area P may regulate accurate and suitable tongue and jaw movements during mastication depending on cortical processing. PMID- 16338827 TI - Factors contributing to the integration of textural qualities: evidence from virtual surfaces. AB - Two experiments involving indirect touch were carried out to explore the relationships among perceptual dimensions of haptically examined surfaces. Subjects in both experiments used a stylus to evaluate the properties of virtual surfaces created by a force-feedback device; four surface properties (resistance to normal force, coefficient of friction, texture scale, and vibration amplitude) were manipulated in various combinations. In Experiment 1, the extent to which there was a one-to-one relationship between specific stimulus properties and perceptual qualities ("perceptual separability") was evaluated. A substantial failure of separability was demonstrated, with friction tending to be more separable from the other properties than they were from one another. The pattern of results suggests that the amount of measured separability depends crucially on the way stimulus properties are defined (e.g., force versus displacement). In Experiment 2, surfaces with known perceptual properties were used to study the metric(s) of the relevant perceptual space. By specifying the perceptual, rather than the stimulus, coordinates of the surfaces, it was possible to bypass issues of perceptual separability. For surfaces of equal friction, a Euclidean metric captured the results (r(2) = 0.75) more effectively than a city-block metric did; neither metric did well when differences in friction were involved. The fact that unlike stickiness-hardness, roughness, and perceived vibration intensity are all increasing functions of surface-normal forces may facilitate their integration into a Euclidean space, in both direct and indirect touch. PMID- 16338828 TI - Sex differences in nociceptive withdrawal reflex and pain perception. AB - Experimentally induced pain often reveals sex differences, with higher pain sensitivity in females. The degree of differences has been shown to depend on the stimulation and assessment methods. Since sex differences in pain develop anywhere along the physiological and psychological components of the nociceptive system, we intended to compare the nociceptive flexion reflex (NFR) as a more physiological (spinal) aspect of pain procession to the verbal pain report of intensity and unpleasantness as the more psychological (cortical) aspect. Twenty female and twenty male healthy university students were investigated by use of nociceptive flexion reflex threshold (staircase method) after electrical stimulation of the N. suralis. Furthermore, we assessed supra-threshold reflex responses (latency, amplitude and area) by applying 10 stimuli 5 mA above reflex threshold. Following each stimulation, the subjects provided pain ratings of intensity and unpleasantness on a visual analogue scale. Females exhibited marked lower nociceptive flexion reflex thresholds than males, while the supra-threshold reflex response tailored to the individual reflex threshold did not show any significant differences. The verbal pain ratings, corrected for NFR threshold, were not found to differ significantly. The large sex differences in nociception that were present in NFR threshold but not in the pain ratings corroborate the hypothesis that spinal processes contribute substantially to sex differences in pain procession. PMID- 16338829 TI - Topography of whisking II: interaction of whisker and pad. AB - The peripheral effector system mediating rodent whisking produces protraction/retraction movements of the whiskers and translation movements of the collagenous mystacial pad. To examine the interaction of these movements during whisking in air we used high-resolution, optoelectronic methods for two dimensional monitoring of whisker and pad movements in head-fixed rats. Under these testing conditions (1) whisker movements on the same side of the face are synchronous and of similar amplitude; (2) pad movements exhibit the characteristic 'exploratory' rhythm (6-12 Hz) of whisking but their movements often have a low frequency (1-2 Hz) component; (3) Pad movements occur in both antero-posterior and dorso-ventral planes but there are considerable variations in the amplitude and topography of movement parameters in the two planes. We conclude that (a) both whisker and pad receive input from a common central rhythm generator; (b) differences in whisker and pad amplitude and topography probably reflect differences in the biomechanical properties of the structures receiving that input; (c) pad movements make a significant contribution to the kinematics of whisking behavior and (d) the two-dimensional nature of pad translation movements significantly increases the rat's flexible control of its mobile sensor. PMID- 16338830 TI - Somatosensory areas S2 and PV project to the superior colliculus of a prosimian primate, Galago garnetti. AB - As part of an effort to describe the connections of the somatosensory system in Galago garnetti, a small prosimian primate, injections of tracers into cortex revealed that two somatosensory areas, the second somatosensory area (S2) and the parietal ventral somatosensory area (PV), project densely to the ipsilateral superior colliculus, while the primary somatosensory area (S1 or area 3b) does not. The three cortical areas were defined in microelectrode mapping experiments and recordings were used to identify appropriate injection sites in the same cases. Injections of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) were placed in S1 in different mediolateral locations representing body regions from toes to face in five galagos, and none of these injections labeled projections to the superior colliculus. In contrast, each of the two injections in the face representation of S2 in two galagos and three injections in face and forelimb representations of PV in three galagos produced dense patches of labeled terminations and axons in the intermediate gray (layer IV) over the full extent of the superior colliculus. The results suggest that the higher-order somatosensory areas, PV and S2, are directly involved in the visuomotor functions of the superior colliculus in prosimian primates, while S1 is not. The somatosensory inputs appear to be too widespread to contribute to a detailed somatotopic representation in the superior colliculus, but they may be a source of somatosensory modulation of retinotopically guided oculomotor instructions. PMID- 16338831 TI - Secondary heat hyperalgesia detected by radiant heat stimuli in humans: evaluation of stimulus intensity and duration. AB - Diverging observations on secondary hyperalgesia to heat stimuli have been reported in the literature. No studies have investigated the importance of heat stimulus intensity and duration for the assessment of secondary heat hyperalgesia. The present study was designed to investigate systematically (1) if pain sensitivity to radiant heat stimuli (focused Xenon light) is altered in the area of secondary punctuate hyperalgesia induced by intradermal injection of capsaicin and (2) if heat stimulus duration and intensity had an influence on the ability to detect secondary heat hyperalgesia. Pain ratings to radiant heat stimuli from a focused xenon lamp were assessed within the area of secondary punctuate hyperalgesia in fifteen volunteers before and after intradermal injection of capsaicin. The stimulus conditions were systematically varied between three intensity levels (0.8, 1.0 and 1.2 x heat pain threshold (PT)) and four duration steps (200, 350, 500 and 750 ms). The present study shows that long duration (350-750 ms) and low intensity (0.8 and 1.0 x PT) radiant heat stimuli were adequate to detect secondary heat hyperalgesia. PMID- 16338832 TI - Persistence in Staphylococcus aureus bacteremia: incidence, characteristics of patients and outcome. AB - Staphylococcus aureus bacteremia often persists. The reasons for persistence and its outcome are poorly defined. We conducted a prospective-observational study among 245 consecutive S. aureus (MRSA: n=125; MSSA: n=120) bacteremias (>or=1 positive blood cultures (BC)) among 234 adults (18-103-y-old; median=59 y) hospitalized during 1 January 2002-31 December 2002 at a 600-bed teaching hospital. Measurements included bacteremia duration, complication-rate (metastatic infection, relapse or attributable mortality) and outcome. Bacteremia duration was measured based on follow-up BC among 193 patients and estimated based on symptoms resolution in the rest. Measured (1-59 d; median=2) and estimated (median=1 d) duration correlated (r=0.885) though positive follow-up BC was often detected without fever (57/105 patients, 54.3%). Persistence (defined as bacteremia for >or=3 d) was noted in 84 cases (38.4%). Complication-rate increased steadily with bacteremia duration (6.6%, 24.0% and 37.7% in bacteremia for 1-2, 3 and >or=4 d, respectively; p=0.05). Cox regression analysis revealed that bacteremia duration correlated positively with endovascular sources (p=0.006), vancomycin treatment (p=0.016), cardiovascular prosthesis (p=0.025), metastatic infections (p=0.025) and diabetes (p=0.038). It is concluded that persistent bacteremia is a feature of S. aureus infection, irrespective of oxacillin susceptibility, associated with worse outcome. Risk factors include endovascular sources, cardiovascular prosthesis, metastatic infections, vancomycin treatment and diabetes. Patients at risk may benefit from novel treatment strategies. PMID- 16338833 TI - Sterile abscess: a surprise diagnosis? AB - We present the case of an otherwise healthy female hairdresser of Brazilian origin who started to have pain and swelling in her left arm. An antecubital abscess was surgically treated at another institution and there was good initial wound healing. Swelling then recurred and fistulae appeared in the scar. Our diagnostic workup revealed an isolated intramuscular tuberculous abscess, which was successfully treated by an antituberculous drug regimen. PMID- 16338834 TI - Validating and updating a prediction rule for neurological sequelae after childhood bacterial meningitis. AB - Recently, a prediction rule for developing neurological sequelae after childhood bacterial meningitis was developed on a small derivation set. Before implementing in practice a prediction rule must first be tested in new patients (external validation). Our aim was to study the external validity of this rule and, if necessary, to update the rule. The prediction rule was tested on newly available data (validation set) by assessing the rule's calibration and discrimination. We updated the prediction rule by adding extra predictors and re-estimating the regression coefficients of the original predictors in the combined datasets. The rule showed poor agreement between predicted risks and observed frequencies. The ROC area was 0.65 (95% CI 0.57-0.72), which was statistically significantly lower than in the derivation set (0.87 (0.78-0.96)), p-value<0.01. The updated prediction rule showed adequate performance in the combined data sets; the ROC area was 0.77 (95% CI 0.72-0.82). Further study of the generalizability of this updated rule may stimulate application in clinical practice. PMID- 16338835 TI - Broad-range real time PCR and DNA sequencing for the diagnosis of bacterial meningitis. AB - Rapid aetiological diagnosis of bacterial meningitis is crucial for the early targeting of antimicrobial and adjuvant therapy. Broad-range polymerase chain reaction (PCR) targeting the 16S rRNA gene allows aetiological diagnosis of bacterial meningitis when applied to cerebrospinal fluid (CSF). We assessed the additional diagnostic effect of applying a novel broad-range real time PCR and subsequent DNA sequencing to culture, microscopy, and broad-range conventional PCR on CSF in patients with suspected bacterial meningitis. Broad-range conventional PCR and broad-range real time PCR with subsequent DNA sequencing were applied to 206 CSF specimens collected consecutively from 203 patients aged 6 d to 86 y. Patients' charts were reviewed for clinical information. 17 pathogens were identified by PCR and DNA sequencing or culture. Three specimens were negative by culture but positive by broad-range real time PCR. Three specimens were positive by culture but negative by broad-range real time PCR. Compared with culture, the sensitivity of broad-range real time PCR was 86%, and the specificity 98%. Conventional PCR resulted in a sensitivity of 64% and specificity of 98%. Broad-range real time PCR was generally comparable to culture of CSF and may be a useful supplement, particularly when antimicrobial therapy has been administered. Broad-range real time PCR was more sensitive than broad range conventional PCR and microscopy. PMID- 16338836 TI - The use of piperacillin/tazobactam (in association with amikacin) in neonatal sepsis: efficacy and safety data. AB - Piperacillin/tazobactam (P/T) has been used in NICU since 1999 as part of the empirical treatment for presumed sepsis. We studied the microbiological and clinical efficacy and safety of P/T use in newborns with sepsis, using a retrospective analysis of medical records of all newborns treated with P/T (in association with amikacin) during 1999-2003. P/T plus amikacin was used for 353 episodes of presumed sepsis occurring in 252 newborns: 105 episodes occurred in 100 newborns treated for 5-14 d while in 248 (70%) episodes the treatment was discontinued after 2-3 d. 123 pathogens were isolated from the blood cultures (40 in the 5-14 d group and 83 in those treated for 2-3 d only). There were 56 Gram negative, 55 Gram-positive and 12 Candida spp. isolates. Klebsiella spp. (40%) and E. coli (30%) were the most commonly isolated Gram-negative pathogens; coagulase-negative Staphylococcus (CONS) represented 95% of the Gram-positive pathogens isolated. Pathogen eradication was achieved within 48-72 h of therapy in 93% Gram-negative organisms. There were no clinical, laboratory or cranial ultrasound adverse effects associated with P/T use. We conclude that P/T in association with amikacin is microbiologically and clinically efficacious and safe in the treatment of sepsis in newborns. PMID- 16338837 TI - Isolated central nervous system histoplasmosis in immunocompetent hosts: a series of 11 cases. AB - Histoplasmosis of the central nervous system occurs in a significant percentage of patients with Histoplasma capsulatum infection, but has usually been described in association with immunosuppression and/or disseminated histoplasmosis. We aim to review the clinical and laboratory features of isolated histoplasmosis of the central nervous system in the immunocompetent host by presenting a series of 11 cases with this condition. Most of these patients presented with headache, meningeal irritation signs and mental status changes, comprising a somewhat different picture from that described in immunosuppressed patients. Moreover, almost all patients had signs of ventricular dilatation in neuroimaging studies, and 8 of the 11 patients had a ventriculoperitoneal shunt at the time of diagnosis, suggesting hydrocephalus to be an important feature of this condition and/or the possibility of shunt infection by the fungus. Immunodiffusion analysis of the cerebrospinal fluid appeared to be the most efficient way to reach the diagnosis and should be considered in immunocompetent patients with chronic lymphocytic meningitis, especially in those who have ventricular shunt or live in endemic areas of Histoplasma capsulatum. PMID- 16338838 TI - Epidemiological study on vaginal Candida glabrata isolated from pregnant women. AB - Candida glabrata was the second most frequently occurring fungus and the dominant non-albicans species caused candidal vulvovaginitis. We used a randomly amplified polymorphic DNA method to compare genotypes of vaginal C. glabrata from pregnant women with or without clinical symptoms. The aims of our study were to define the prevalence of C. glabrata during pregnancy and identify the relationship between genotypes of C. glabrata and the clinical symptoms. A total of 50 C. glabrata strains were isolated, identified, and genotyped from 628 pregnant women. The prevalence of C. glabrata during pregnancy was 7.96% (50/628). C. glabrata was detected in 25.12% (50/199) of all Candida isolates. 17 unique genotypes were generated by RAPD and the mean SAB value of all isolates was 0.891+/-0.002. All results show that the genotypes of vaginal C. glabrata isolated from pregnant women were highly similar but non-identical. Compared to those of symptomatic patients in the same trimester and asymptomatic patients in different trimesters, C. glabrata isolated from the asymptomatic patients in the first trimester had more genotypical similarities. Genotypical similarities of C. glabrata were related to clinical signs and symptoms, to some degree. PMID- 16338839 TI - Unexpected high prevalence of IgG-antibodies to hepatitis E virus in Swedish pig farmers and controls. AB - Hepatitis E virus (HEV) infections are responsible for large waterborne outbreaks in developing countries. Sporadic cases in the developed world are mainly imported via immigrants and travellers from endemic areas. HEV has been suggested to be a zoonotic infection where pigs may be an important reservoir for the disease and specific swine strains of HEV have been identified which can infect also humans. The aim of this study was to analyse if Swedish pig farmers are more exposed to HEV than persons with other occupations. A total of 115 male pig farmers aged 40-60 y and 108 age- and geographically- matched control subjects were tested for IgG anti-HEV antibodies. No statistical difference in anti-HEV prevalence was noted between pig farmers (13.0%) and control subjects (9.3%). The prevalence of anti-HEV antibodies in the pig farmers and controls was higher than that previously reported among other populations in Europe (<1-9%). Further studies are needed to elucidate the routes for infection of indigenous HEV and if sub-clinical infections with pig associated HEV strains occur in Sweden. PMID- 16338840 TI - Clostridium cadaveris bacteraemia: two cases and review. AB - Clostridium cadaveris is a strict anaerobic Gram-positive rod that is the most prominent bacterium during the decay of dead bodies. We present 2 rare cases of bacteraemia with C. cadaveris. The source of both infectious episodes was most probably of gastrointestinal origin. PMID- 16338841 TI - Intracerebral haemorrhage as a rare complication of HSV-1 meningoencephalitis: case report and review of the literature. AB - We present a case of herpetic meningoencephalitis confirmed by PCR in a 22-y-old male, with accompanying appearance of a large intracerebral haematoma as a complication. Despite the impressive imaging findings, the final outcome of the patient's progress was favourable. PMID- 16338842 TI - Gas within the liver and polymicrobial bacteraemia due to colovenous fistula: two cases in one month. AB - Two cases presented at our hospital within 1 month with polymicrobial bacteraemia and gas within the liver shown on CT scan. Both transpired to have colovenous fistula due to diverticulitis. Because of our awareness of the first case, the colovenous fistula of the second case was identified rapidly. As surgical treatment is probably essential for this condition, the possibility of colovenous fistula should be borne in mind in patients with gas within the liver, especially if they have bacteraemia. PMID- 16338843 TI - Pseudomonas fluorescens infections in clinical practice. AB - Pseudomonas fluorescens was isolated from an elderly immunocompromized patient with fever. Treatment with ceftazidime was successful, after empirical therapy failed. Pseudomonas fluorescens is 1 of the less virulent members of the Pseudomonadaceae family. The epidemiology of the infection and the difficulties in isolation and susceptibility assessment are further discussed. PMID- 16338844 TI - Cutaneous melioidosis in a Swedish tourist after the tsunami in 2004. AB - A tourist from Sweden developed cutaneous melioidosis after the tsunami in Thailand on 26 December 2004. Melioidosis is a severe, chronic infection which is endemic in Thailand and is caused by Burkholderia pseudomallei. Persons with traumatic injuries inflicted by the tsunami have increased risks of being infected by B. pseudomallei and melioidosis should be suspected if abscesses of the skin or inner organs develop in the months or years after the trauma. PMID- 16338845 TI - Clinical improvement of psoriasis in an AIDS patient effectively treated with combination antiretroviral therapy. AB - Psoriasis has been classified as a T-cell-mediated inflammatory disease. The paradoxical exacerbation of psoriasis in AIDS has not been fully explained. We describe a case of a 45-y-old male with AIDS whose coincidental psoriasis resolved following antiretroviral therapy. Dramatic improvement was seen 4 weeks after combination antiretroviral therapy including enfuvirtide was started. In advanced HIV disease psoriasis could represent a clinical index of progression of HIV disease. Several pathogenetic factors involved in the clinical manifestations of psoriasis in the HIV-infected population are discussed. PMID- 16338846 TI - A fatal case of severe immunodeficiency associated with disseminated Merulius tremellosus infection. AB - We report a fatal case of severe immunodeficiency associated with infection caused by the white rot fungus, Merulius tremellosus. This is the first report of Merulius tremellosus infection in humans. To improve clinical management, awareness and diagnostics of uncommon fungal infections in immunodeficient patients are important. PMID- 16338847 TI - Thermal medicine, heat shock proteins and cancer. PMID- 16338849 TI - How do cells respond to their thermal environment? AB - Changes in growth temperature induce both activating and inactivating responses from cells, with the magnitude of the temperature change being among the factors that influence which type of response dominates. Aside from upregulated enzyme activity, induction of thermotolerance is the most widely studied and best understood activating response that cells exhibit following heat shock. Inactivating responses to heat shock that are of biomedical interest include heat radiosensitization and cytotoxicity. Interestingly, the activation energy for inducing thermotolerance, heat cytotoxicity, and radiosensitization all fall within a similar range of 120-146 kcal per mole. The relatively high activation energy for each of these responses suggests that they all involve a heat-induced molecular transition as a trigger, and several lines of research suggest strongly that protein denaturation is the common transition that triggers all three responses. Low levels of protein denaturation are sufficient to attract the 90 kDa heat shock protein (HSP90) such that it frees up heat shock factor 1, which then trimerizes to form an active transcription factor that upregulates expression of heat shock proteins. Upregulation of heat shock proteins and other heat-induced events result in the development of thermotolerance, which protects cells from subsequent exposure to heat shock and other stresses. A more severe heat shock increases protein denaturation proportionately and leads to aggregation of both denatured and native proteins. This results in inactivation of protein synthesis, cell cycle progression, and DNA repair processes such that cells either die or are sensitized to radiation and other cytotoxic events. The ultimate fate of cells following a heat shock depends upon the summation of the activation and inactivation events that are induced, which appears to be governed by the resultant magnitude of protein denaturation and aggregation. Treatments that stabilize cellular proteins against denaturation and aggregation reduce the magnitude of inactivating responses while increasing that of activating responses for a given heat shock (time at temperature), while treatments that sensitize proteins to denaturation and aggreation have the converse effect. These findings support the conclusion that the determinant of the cellular response to heat shock is the amount of heat-induced protein denaturation and aggregation and not the time at temperature. PMID- 16338850 TI - Role of HSPs and telomerase in radiotherapy. AB - The inability of radiotherapy to control tumour growth is still a daunting clinical problem leading to failure of the overall treatment regimens. The fundamental question is; could tumour cells be specifically sensitized to ionizing radiation (IR) by heat or factors exclusively expressed in tumour cells? One such factor, expressed in most tumours and silent in somatic cells, is telomerase. Biochemical and genetic studies have established an association between telomere maintenance and extended life span of human cells mediated through the expression of the catalytic sub-unit of telomerase (hTERT). Because of this, telomerase is an attractive target for inhibition in anti-cancer therapy. Telomeres are maintained by telomerase and hTERT interacts with heat shock protein (HSP) chaperones. This review will focus on the possible role of HSPs and telomerase in sensitizing tumour cells and, thus, enhancing the potential of targeted radiotherapy. PMID- 16338851 TI - Inhibiting induction of heat shock proteins as a strategy to enhance cancer therapy. AB - Cancer treatments that incorporate thermal therapy and some systemic therapies induce the production of heat shock or stress proteins. The induced heat shock proteins could lessen the effect of the therapy by inhibiting apoptotic signaling and by acting as molecular chaperones to prevent irreversible cellular damage. Strategies that prevent the induction of heat shock proteins would result in more apoptosis and necrosis, improving the cancer therapy. This paper briefly reviews cancer therapies that induce the stress response, and proposes strategies to reduce the stress response. PMID- 16338852 TI - Inducers and co-inducers of molecular chaperones. AB - Molecular chaperones, which are mostly heat- or stress-induced proteins (HSPs), not only regulate various cellular functions such as protein folding, refolding of partially denatured proteins, protein transport across membranes, cytoskeletal organization, degradation of disabled proteins, and apoptosis, but also act as cytoprotective factors against deleterious environmental stresses. Recent studies indicated that moderate overexpression of molecular chaperones could confer cells and tissues stress tolerance and provide beneficial effects on various pathological states associated with protein misfolding and protein aggregation. Mild heat shock, transfection of HSP genes, and some chemical compounds are the major means of overexpression of molecular chaperones. In this review, we summarize recent studies of chemical compounds that could induce or enhance the expression of molecular chaperones or HSPs. PMID- 16338853 TI - How is the immune response affected by hyperthermia and heat shock proteins? AB - There is growing body of evidence linking the cellular response to heat stress with the response of the immune system to cancer. The anti-tumor immune response can be markedly enhanced by treatment with hyperthermia particularly in the fever range. In addition, the heat shock proteins (hsp) which are produced in abundant quantities in cells exposed to heat are potent immune modulators and can lead to stimulation of both the innate and adaptive immune responses to tumors. Immunostimulation by hyperthermia involves both direct effects of heat on the behavior of immune cells as well as indirect effects mediated through hsp release. In addition, the hsp can be deployed as components of antitumor vaccines in protocols that do not include hyperthermia. Understanding these process may permit the effective deployment of hyperthermia and hsp based vaccines in tumor treatment. PMID- 16338854 TI - Current ideas about applications of heat shock proteins in vaccine design and immunotherapy. AB - Heat shock proteins (HSPs), as molecular chaperones, perform critical functions in maintaining cell homeostasis. Certain HSPs in extra-cellular milieu are capable of modulating innate and adaptive immunity due to their ability to chaperone polypeptides and to interact with the host's immune system, particularly professional antigen presenting cells (APCs). This review summarizes the immunomodulating functions of HSPs and their potential applications in cancer immunotherapy. PMID- 16338855 TI - Thermal regulation of lymphocyte trafficking: hot spots of the immune response. AB - Lymphocytes use extensive vascular networks to traffic to various destinations in the body, including lymphoid organs and extra-lymphoid tissues. This discussion will focus on the emerging evidence that thermal stress regulates the traffic signals that direct the exit of lymphocytes from the vascular freeway. This issue is particularly relevant to T cell-based cancer immunotherapy where delivery of immune effector lymphocytes to neoplastic lesions depends on their extravasation across tumour micro-vessels. Although tumours are frequently highly vascularized by vessels that are competent to support blood flow, the tumour micro-environment has been characterized as non-permissive to lymphocyte extravasation. This may lead to a scenario where limited leukocyte infiltration at tumour sites correlates with a poor prognosis. These observations support the thesis that adjuvant strategies that promote trafficking of tumour-reactive cytolytic leukocytes to tumour sites have the potential to improve the efficacy of immune based cancer therapy. PMID- 16338856 TI - Evolving technology for thermal therapy of cancer. AB - This paper is intended as a succinct review of technology used for clinical hyperthermia therapy for cancer, as culled from a presentation at the special workshop on Thermal Medicine, Heat Shock Proteins, and Cancer at the Society for Thermal Medicine conference in Spring 2005. Following a brief overview of thermal therapy treatment options and available mechanisms for heating tissue, the paper focuses on the evolution of equipment from basic single element heating devices of the early 1980s to adjustable multi-element heating devices currently in use or in final stages of development. Representative devices from the past, present and near future are cited for further investigation by the interested reader. The paper concludes with a summary of general trends in the evolution of clinical hyperthermia techniques and a statement of current challenges remaining for the field. PMID- 16338857 TI - Thermal ablation and high-temperature thermal therapy: overview of technology and clinical implementation. AB - High-temperature hyperthermia or thermal therapy is being applied for destruction of cancerous tissue, eradication or reduction of benign tumours and targeted tissue modification and remodelling. Many of these high-temperature technologies provide a minimally-invasive alternative with lower morbidities compared to the traditional surgical procedures. The effects of high-temperature thermal exposure on tissues, examples of heating technology and procedures of clinical practice related to high-temperature thermal therapy are reviewed. This brief review encompasses interstitial, endocavity, intraluminal and external applications of RF, microwave, ultrasound, laser and thermal conduction energy sources. The technology is prevalent and in various levels of advancement, with the move toward more spatially-accurate and controllable heating systems combined with image-guidance and treatment verification warranted, especially for the treatment of cancer. PMID- 16338858 TI - Thermal tumour ablation: devices, clinical applications and future directions. AB - Tumour ablation is clinically applied mainly for non-operable liver tumours, with increasing application to other organ sites like kidney, lung, adrenal gland and bone. Most current devices use radiofrequency (RF) current to heat tumour tissue surrounding the applicator, which is introduced into the tumour under imaging guidance. Tissue temperatures in excess of 100 degrees C are achieved, with cell death due to coagulative necrosis occurring above 50 degrees C. Limitations of current ablation devices include inadequate imaging, limited size of coagulation zone and reduced performance next to large vessels. This paper reviews current interstitial RF and microwave devices, clinical applications and future research directions in the field of high-temperature tumour ablation. PMID- 16338859 TI - Implications of increased tumor blood flow and oxygenation caused by mild temperature hyperthermia in tumor treatment. AB - In many past clinical studies in which hyperthermia enhanced the efficacy of radiotherapy, the tumor temperatures could be raised only to 40-42 degrees C range in most cases. The heat-induced cell death, cellular radiosensitization, and vascular damage induced by such mild temperature hyperthermia (MTH) are likely to be insignificant despite the increased response of tumors to radiotherapy. Heating rodent tumors at 40-42 degrees C was found to cause an enduring increase in blood flow and oxygenation in the tumors. Recent studies with canine soft tissue sarcoma and human tumor clinical studies also demonstrated that MTH improves tumor oxygenation, and enhances response of the tumors to radiotherapy or chemoradiotherapy. The increased blood flow and vascular permeability caused by MTH may also improve the delivery of various therapeutic agents such as chemotherapy drugs, immunotherapeutic agents and genetic constructs for gene therapy to tumor cells. MTH as a means to potentiate the efficacy of radiotherapy and others warrants further investigation. PMID- 16338860 TI - The heat shock response: role in radiation biology and cancer therapy. AB - Since prehistoric times, elevated temperatures have been used to treat cancer in a variety of forms. In modern times (the last 40 years) efforts have concentrated on combining heat with other anti-tumour modalities, principally ionizing radiation and some chemotherapeutic drugs. Despite the emphasis on combined therapy, rodent data relating to heat sensitivity and thermal tolerance development assumed principal importance. These considerations suggested treating at 43 degrees C as a target temperature and fractionation schemes emphasizing thermal tolerance avoidance. Concomitantly crucial data on heat-induced tumour reoxygenation and its temperature dependence were largely ignored. In reality these were unrealistic and undesirable goals. The preponderance of evidence now suggests that lower temperatures (40-42 degrees C) administered more frequently, optimally immediately before and during each administration of ionizing radiation, are likely to yield optimal results. Factoring in trimodality therapy and other combinations of chemotherapeutic drugs will require some modifications of such fractionation schemes. PMID- 16338861 TI - Re-setting the biologic rationale for thermal therapy. AB - This review takes a retrospective look at how hyperthermia biology, as defined from studies emerging from the late 1970s and into the 1980s, mis-directed the clinical field of hyperthermia, by placing too much emphasis on the necessity of killing cells with hyperthermia in order to define success. The requirement that cell killing be achieved led to sub-optimal hyperthermia fractionation goals for combinations with radiotherapy, inappropriate sequencing between radiation and hyperthermia and goals for hyperthermia equipment performance that were neither achievable nor necessary. The review then considers the importance of the biologic effects of hyperthermia that occur in the temperature range that lies between that necessary to kill substantial proportions of cells and normothermia (e.g. 39-42 degrees C for 1 h). The effects that occur in this temperature range are compelling-including inhibition of radiation-induced damage repair, changes in perfusion, re-oxygenation, effects on macromolecular and nanoparticle delivery, induction of the heat shock response and immunological stimulation, all of which can be exploited to improve tumour response to radiation and chemotherapy. This new knowledge about the biology of hyperthermia compels one to continue to move the field forward, but with thermal goals that are eminently achievable and tolerable by patients. The fact that lower temperatures are incorporated into thermal goals does not lessen the need for non-invasive thermometry or more sophisticated hyperthermia delivery systems, however. If anything, it further compels one to move the field forward on an integrated biological, engineering and clinical level. PMID- 16338862 TI - Histologic study of the tunica albuginea of the penis and mode of cavernosus muscles' insertion in it. AB - The tunica albuginea (TA) is the outer covering of the corpora cavernosa (CCs) and corpus spongiosum (CS) of the penile shaft. The different histoanatomical patterns of the TA, as well as the mode of attachment of the ischio-/bulbo cavernosus muscles (ICM, BCM) to the TA, were studied, aiming at elucidating their role in the mechanism of erection. Twenty-eight cadaveric specimens (18 adults, 10 neonatal deaths) were studied morphologically and histologically after staining with hematoxylin and eosin and Verhoeff-van Gieson stains. The TA consisted in 20 specimens of 2 layers: inner circular and outer longitudinal, in 6 specimens of 3 layers: inner circular, longitudinal and outer circular, and in 2 of only one longitudinal layer. The CS TA was formed of one layer of longitudinal fibers. The mode of cavernosus muscles insertion into the TA revealed 3 patterns. The conjoint BCM and ICM formed a fibrous belt over the CCs in 18 specimens, a muscular belt in 6 and no belt in 4. The functional role of the variations in the TA morphological structure is not exactly known. We hypothesize that the 3-layered TA gives more penile rigidity than the double and single layers. Considering the type of cavernosus muscles insertion into the TA, it appears that the fibrous belt exerts more CC compression than the other 2 types of insertion. PMID- 16338863 TI - Growing concern over the safety of using mobile phones and male fertility. AB - There are growing concerns about the possible hazards of electromagnetic waves emitted by mobile phones on human health. One of the biggest concerns is their possible association with increased risk of cancer and their possible effects on cellular DNA. Electromagnetic waves can inflict their results through both thermal and non-thermal effects. There are many animal studies that show that electromagnetic waves have a wide range of damaging effects on the male reproductive system and sperm parameters. However, similar studies are quite limited in humans, and the results of animal studies should be interpreted with caution when considering their application to humans. Large controlled studies are required before confirming such possible effects on male fertility. PMID- 16338864 TI - Screening of FSH receptor gene mutation (C566T) in azoospermic men in Japan. AB - To study the genetic mutation, which could cause problems in spermatogenesis, we screened the point mutations of the FSH receptor gene (C566T) in idiopathic azoospermic men in Japan. We performed mutational analysis of the FSH receptor in 54 Japanese patients diagnosed for azoospermia with Sertoli cell-only (SCO) syndrome (n = 33), hypospermatogenesis (n = 11) and maturation arrest (n = 10). For mutation screening of the FSH receptor, polymerase chain reaction (PCR) amplification from genomic DNA with flanking intronic primers were used. On BsmI digestion, all patients demonstrated homozygous, normal exon 7 alleles with 51 and 27 bp fragments. The absence of any 78 bp fragments demonstrated that no heterozygous or homozygous mutant alleles were present in any patients. None of the 54 patients showed a C566T FSH receptor mutation. We could not confirm that the genomic mutation of the FSH receptor (C566T) is a common cause in Japanese azoospermic patients. PMID- 16338865 TI - Does smoking change the efficacy of combination therapy with vitamin E and colchicines in patients with early-stage Peyronie's disease? AB - This study was conducted to find out if smoking has an effect on the results of combination therapy with vitamin E and colchicines in patients with early-stage Peyronie's disease (PD). A total of 58 potent patients suffering from early-stage PD were included in the study (mean age 47.3 years, range 25-73 y). The time from onset of the disease was <6 months and no patient had ED. The patients with severe fibrotic or calcified plaques were not included in the study. Of the patients, 36 were smokers (Group 1) and 22 were non-smokers (Group 2). All the patients received vitamin E (800 IU daily) and colchicines (1 mg daily) for 6 months. Follow-up ranged from 5 to 13 months (mean 10.3 m). The combination therapy was effective and well tolerated in both groups. There were no significant differences between the two groups according to age, disease duration, related disease (diabetes, hypertension, hypercholesterolemia, and hypertriglyceridemia), plaque sizes, and plaque numbers. The resolution in pain and increase in penile curvature and plaque size were similar rates in both groups (p > 0.05), while decrease in penile curvature and plaque size were higher in Group 2 (p < 0.05). No patient discounted the therapy due to side effects. The oral combination therapy with vitamin E and colchicines appears to be an effective procedure in patients with early-stage PD and smoking may have worsening effects on the treatment results. PMID- 16338866 TI - Measurement of electrolyte concentrations in expressed prostatic secretion and urine from patients with chronic prostatitis and its implications. AB - This study was aimed at measuring concentration of electrolytes, especially K+ in expressed prostatic secretion (EPS) and urine from patients with chronic prostatitis. The concentration of potassium, sodium, chloride, calcium in EPS and urine of 31 controls and 79 patients with prostatitis were measured and analyzed. There was no significant difference in the concentrations of potassium, sodium, chloride and calcium between the patients and the controls. Among the patients treated effectively, potassium concentration was 40.66 +/- 17.10 mmol/l before treatment and 33.42 +/- 17.27 mmol/l after treatment. While among the patients treated ineffectively, potassium concentration was measured as 37.57 +/- 16.93 mmol/l and 50.66 +/- 18.77 mmol/l before and after treatment respectively. The concentrations of electrolytes in prostatic fluid varied greatly between individuals. Potassium concentration in EPS decreased significantly after treatment among the patients with obvious treatment effectiveness, while increased among those who failed the treatment. EPS potassium concentration was also found to be lower in patients with pain than those without pain. No significant difference was found between the normal group and the no-pain patients. PMID- 16338867 TI - Treatment with prednisolone of hormone-refractory prostate cancer. AB - Fifteen patients 60 to 80 years old (a mean of 72 years) with hormone-refractory prostate cancer were treated with low dose prednisolone. All patients had previously undergone hormone therapy. Prostate specific antigen (PSA) values decreased in 11 cases (73%), of which 4 had PSA decreases of 50% or greater. Serum levels of DHEAS significantly decreased at 4 and 8 weeks after treatment (both intervals were p < 0.05 vs pretreatment). Of 8 patients with bone metastasis evaluation, 2 (25%) showed improvement of the lesion. In 5 patients (33%), relief of pain was observed one month after starting prednisolone. The one year survival rate was 58%. The side effects were mild and manageable in an outpatient clinic. PMID- 16338868 TI - A follow-up expanded study of the correlation of sperm velocity in seminal plasma and offspring gender. AB - A preliminary study reported finding higher sperm velocity in seminal plasma in males of partners that conceived female offsprings. The null hypothesis was that sperm velocity was not related to the offspring gender. The objectives were: (a) to expand the previous study, and (b) to correlate offspring gender results with motility parameters determined through the computer-aided sperm analyzer (CASA) system. In combined fresh and frozen cycles (N = 187), sperm from cases with all female offsprings displayed higher curvilinear (48 +/- 1.0 mu/sec versus male 46 +/- 1.0, P < 0.05) and average path velocities (36 +/- 0.7 mu/sec versus male 34 +/- 0.7, P < 0.01). A criteria of less than 30 mu/sec or over 41 mu/sec average path velocity predicted 73 or 72% of the male or female offspring cases, respectively. A curvilinear velocity of less than 49 mu/sec or over 55 mu/sec predicted 58 or 59 % of the male or female offspring cases, respectively. Semen viscosity reflected in sperm velocity was linked to predominantly male or female sperm populations. Paracrine signals from the gender-skewed sperm precursor populations controlling viscosity merit further exploration. PMID- 16338869 TI - Evidence favoring sperm selection over sperm competition in the interaction between human seminal plasma and sperm motility in vitro. AB - The effect on sperm motility of sperm-sperm and sperm-seminal plasma interactions was studied among homologous and heterologous sperm. There were no significant interactions between sperm in vitro, but it was found that seminal plasmas of different donors have different effects on sperm motility, and different sperm react differently to the same seminal plasma. Sperm showed higher motility in a pure physiological solution than when mixed with seminal plasma, even if the plasma and sperm came from the same donor. Various plasma components are responsible for this modulation of sperm motility. It would appear that large numbers of sperm are adaptive, among other things, because they are involved in sperm selection. PMID- 16338870 TI - Ultrastructural and DNA fragmentation analyses in swim-up selected human sperm. AB - Seventeen sperm samples were evaluated by transmission electron microscopy (TEM) before and after swim-up separation. DNA-fragmentation was tested by terminal d UTP nick end labeling (TUNEL) in unselected and selected semen samples, and the results were analyzed in relation to sperm ultrastructural characteristics detected by TEM. A significant improvement in mean numbers and percentages of structurally normal sperm was observed after swim-up selection, corresponding to a significant decrease in the percentage of necrotic and apoptotic sperm, while the percentage of sperm with immature nuclei did not change significantly. TUNEL indicated a significant decrease in chromatin-fragmented sperm after swim-up. Swim up selection based on sperm motility excludes many sperm with ultrastructural evidence of necrosis (absent or reacted acrosome, disrupted chromatin, broken plasma membrane) and apoptosis (misshapen nuclei with marginated chromatin), as confirmed by TUNEL analysis. Nevertheless, immature sperm with elliptical or roundish nuclei, misshapen acrosomes and uncondensed chromatin remain part of fertilizing pool. PMID- 16338871 TI - Effects of phototherapy on newborn rat testicles. AB - Phototherapy is the most widespread treatment for lowering bilirubin concentration in neonates. In the routine, phototherapy has some side effects including skin eruption, fluid loss, abdominal distention, mild hemolysis and mild thrombocytopenia. The aim of the study was to investigate the possible mutagenic and gametocidal side effects of 72 h continuous phototherapy on the rat testicle. We observed decreases in spermatogonia numbers per tubule (S/T values), tubular fertilization index (TFI) and sperm sertoli cell index (SSCI), which are the most reliable methods in estimating future fertility potential, due to sensitivity to phototherapy. The differences between study and control groups for S/T, TFI and SSCI values were statistically significant (p = 0.008, p = 0.02 and p = 0.004, respectively). There were significant differences in seminiferous tubule diameters between the control and study groups (p < 0.005), but no significant difference in DNA index values between the control (0.66 +/- 0.12) and study (0.59 +/- 0.05) groups (p > 0.05). As a conclusion, phototherapy seems to have some side effects on the newborn rat testicle. Further studies with larger groups, designed for investigation of the effects of phototherapy on seminiferous tubules, may give more beneficial results. PMID- 16338872 TI - Effects of testicular fixation on oxidative stress-related parameters in prepubertal rat testis. AB - The goal of orchiopexy is to provide adequate scrotal fixation to prevent recurrent torsion of the testis and the spermatic cord or ascent of the testis, to be achieved with minimal trauma to the testis. Forty-two prepubertal male Sprague-Dawley rats were equally distributed into 6 groups (1 SHAM + 5 Operation). In operational groups, two lateral sutures were placed near the upper and lower pole of the testis passing through the tunica albuginea with 4-zero polyglactin suture in the transverse axis of the testis. At the 1st-3rd-7th-15th and 30th postoperative days, bilateral orchiectomy was performed, then malondialdehyde (MDA), nitrite, superoxide dismutase (SOD) and glutathion peroxidase (GPx) values were measured in testicular tissue homogenates. MDA and nitrite levels increased, while GPx and SOD levels diminished, statistically significant in both ipsilateral and contralateral testicular tissue homogenates. Prepubertal testicular fixation causes overproduction of the lipid peroxidation end product and free radicals while heavy consuming anti-oxidant reserve systems in both ipsilateral and contralateral testicular tissue. These results clearly reveal the extent of the testicular deterioration due to suture fixation technique. PMID- 16338874 TI - Lipoblastoma in a four-year-old African child. AB - A four-year old Sudanese child presented with a growing mass in the medial aspect of the right thigh. The mass appeared during the neonatal period. On clinical examination a diagnosis of lipoblastoma was entertained on the basis of the patient's age and the clinical features of the mass. The tumor was completely excised surgically. The clinical diagnosis of lipoblastoma was confirmed pathologically. Follow-up of the patient for 6 months postoperatively showed no evidence of recurrence. PMID- 16338875 TI - Immunohistochemical expression of endothelial nitric oxide synthase and C-kit in the placenta of complete hydatidiform mole. AB - Complete hydatidiform mole is an abnormal conceptus characterized by hydropic villi accompanied by proliferating trophoblasts. Its pathogenesis is largely unknown. Endothelial nitric oxide synthase is induced by vascular endothelial growth factor and has been implicated in the pathogenesis of preeclampsia and other physiologic conditions in the placenta. C-kit is the tyrosine kinase receptor and is involved in tumor formation elsewhere in the body. Using standard immunohistochemical protocols, we studied the expression of C-kit and endothelial nitric oxide synthase in the placenta of 10 patients with complete hydatidiform mole. Cytoplasmic and nuclear staining with endothelial nitric oxide synthase was identified in the cytotrophoblast and intermediate trophoblast layers in all cases, with high staining in 7/10 and 6/8 cases, respectively. Minimal staining is identified in the syncytiotrophoblast layer. Hofbauer stromal cells were identified in 9 cases and showed low staining intensity in 7/9 cases. Cytoplasmic C-kit staining was diffuse and of low intensity. The cytotrophoblast, the syncytiotrophoblast, intermediate trophoblast, and the stromal cells had low C kit staining intensity in 8/10, 8/10, 7/9, and 5/9 cases. These results indicate that C-kit and endothelial nitric oxide are expressed in the placentas of complete hydatidiform mole and may play a role in the pathogenesis of trophoblastic proliferation in this condition. PMID- 16338876 TI - Cytochrome c oxidase deficiency in a child with isolated myopathy. AB - Cytochrome c oxidase (COX) deficiency is the most commonly recognized respiratory chain defect in childhood. The disease is clinically heterogeneous with phenotypes including Leigh syndrome, hepatic failure and myopathies. COX deficiency has been associated with mitochondrial DNA mutations in COX I, II, and III with large-scale deletions of the mitochondrial genome and with point mutations in mitochondrial tRNA genes. Here we report on a 3.5-year-old girl with a rare type of isolated myopathy due to COX deficiency. PMID- 16338877 TI - Intrathoracic ectopic lobe of liver presenting as pulmonary sequestration. AB - Intrathoracic ectopic lobe of the liver in the presence of a normal intact diaphragm is extremely rare. We report a case of a 13-month-old male initially diagnosed with pneumonia and pulmonary sequestration who was found to have an intrathoracic liver lobe and intact diaphragm. The presence of this condition suggests an event preceding closure of the diaphragm and illustrates the unique potential of pediatric pathology to shed light on human embryology. PMID- 16338878 TI - Squamous cell peritonitis associated with hydrometrocolpos in a multimalformed newborn. AB - We present the case of a 1 day-old newborn with extensive squamous cell peritonitis resulting from vaginal atresia with hydrometrocolpos and squamous cell reflux through the genital system, a combination rarely recognized in the literature. Delivery was preceded by ultrasound diagnosis of oligohydramnios and "large bladder," the latter representing the dilated proximal vagina. Additional findings included paraesophageal hiatus hernia containing squames, urethral atresia, absence of vulva, lung hypoplasia, mild hypoplasia of the corpus callosum, and short umbilical cord with single umbilical artery (SUA). The possible relationship of the SUA with the constellation of infradiaphragmatic malformations is discussed. PMID- 16338879 TI - The usual childhood diseases: forgotten but not gone. AB - With the advent of vaccine protection for many diseases, many of today's practitioners have never seen cases of what were the common childhood diseases. If any of these diseases returns, there is a risk that the practitioner will not recognize it. This article tries to establish why the diseases may return and to describe the diseases with photographs. PMID- 16338880 TI - Empty sella syndrome, panhypopituitarism, and diabetes insipidus. AB - We present an 18-month-old girl with short stature, obesity, panhypopituitarism, diabetes insipidus, and visual defects. Postmortem examination revealed brain atrophy due to a diffuse encephalopathy, numerous calcified neurons in cerebral cortex, deep telencephalic and diencephalic nuclei, diffuse neuronal necrosis in hypothalamic nuclei, moderate atrophy of optic nerves, very thin hypophyseal stalk, and empty sella with the hypophysis compressed to the dorsal aspect of the concavity. Our hypothesis is that the presence of an empty sella in a child with hypophyseal-hypothalamic abnormalities should alert physicians to the existence of hypothalamic lesions secondary to a perinatal insult. We discuss the possible pathogenesis of these findings as well as lines of evidence available in the literature. PMID- 16338881 TI - From the editor. White House Conference on Aging. PMID- 16338882 TI - Practical considerations for maintaining independence among individuals with functional impairment. AB - Social workers, gerontologists, and related health professionals must learn to recognize how aging, disability, and chronic illness affects individuals and their families' physical, psychological, and social functioning in order to develop effective interventions for their clients. This article discusses prevention strategies to help facilitate functional consequences of aging, including chronic illness and disease and offers suggestions for assisting clients to cope and manage the consequences of illness in frail elder adults. PMID- 16338883 TI - Understanding the experience of moving a loved one to a long-term care facility:family members' perspectives. AB - This project was designed to develop an understanding of family members' experiences of moving a loved one to a long-term care facility and to identify ways in which facilities might help ease this process. Twenty-one semi-structured interviews were conducted with family members who had recently moved a relative into one of three long-term care facilities in Southern Ontario, Canada. Several factors appeared to contribute to the overall experience of the move to long-term care and either served to impede or facilitate a positive transition for families. These factors included: the experience during the waiting process, preparation for the move, ease of the actual move, control over decisions, communication throughout the process, support from others, and family and resident perceptions and attitudes towards the move. Easing the difficult aspects of moving a loved one to a long-term care setting can be facilitated with better preparation and support from facilities and community services. PMID- 16338884 TI - Depression and caregiver burden among rural elder caregivers. AB - Family caregivers of older adults frequently experience feelings of burden and depression though they may not come to the attention to health and service providers until they are at a point of crisis. Through a simple screening tool, the Maine Primary Partners in Caring (MPPC) project identified individuals providing care to older adults through rural primary care practices, in order to provide upstream interventions before caregivers were in crisis. This paper describes a sample (n=62) of rural family caregivers identified through their physicians' offices. High levels of caregiver burden and depression were reported. Family support and knowledge of caregiver tasks predicted decreased caregiver burden and depression, while isolation predicted increased caregiver burden. Implications of these results for gerontological social workers are outlined. PMID- 16338885 TI - Use of senior centers as a moderator of stress related distress among Latino elders. AB - Little is known about the moderating effects of community services on psychological distress among Latino elders. This study explores the use of senior centers as an intervention strategy to moderate the harmful impact of stress on psychological distress. The study was conducted using data from the 1988 National Survey of Hispanic Elderly People (N=2,299). To examine the direct and moderating effects of the use of senior centers on the relationship between stress and psychological distress we applied multiple regression analysis to an initial model, a direct effect model, and a moderating model. The results indicated that, under stressful life situations, respondents who used senior centers experienced lower levels of psychological distress than those who did not. Implications of the findings for senior centers and for further research are discussed. PMID- 16338886 TI - Ombudsman program characteristics related to nursing home abuse reporting. AB - The purpose of this research is to determine if there is an association between ombudsman report rates of abuse, gross neglect, and exploitation from 1997 through 2002 and ombudsmen's public and staff education, investigative process, program expenditures, and census demographics. Data were obtained from the National Ombudsman Reporting System. Ombudsman annual reports of nursing home abuse, neglect, and exploitation are representative of that state's individual program. These reports should not be construed to represent all abuse in nursing homes. Physical abuse and resident-to-resident abuse were the highest rates of abuse reported. Census demographics and ombudsman program characteristics were associated with rates of abuse. PMID- 16338888 TI - Same-sex desire and love in Greco-Roman antiquity and in the classical tradition of the West: Introduction. PMID- 16338889 TI - Reconsiderations about Greek homosexualities. AB - Focusing his analysis on (mostly Athenian) vase paintings of the sixth- and early fifth-century and on a handful of texts from the late fifth- and early fourth century (again Athenian), Dover depicted the pederastic relationship of erastes (age 20 to 30) and eromenos (age 12-18) as defined by sexual roles, active and passive, respectively. This dichotomy he connected to other sexual and social phenomena, in which the active/ penetrating role was considered proper for a male adult Athenian citizen, while the passive/penetrated role was denigrated, ridiculed, and even punished. Constructing various social and psychological theories, Foucault and Halperin, along with a host of others, have extended his analysis, but at the core has remained the Dover dogma of sexual-role dichotomization. Penetration has become such a focal point in the scholarship that anything unable to be analyzed in terms of domination is downplayed or ignored. To reduce homosexuality or same-sex behaviors to the purely physical or sexual does an injustice to the complex phenomena of the Greek male experience. From Sparta to Athens to Thebes and beyond, the Greek world incorporated pederasty into their educational systems. Pederasty became a way to lead a boy into manhood and full participation in the polis, which meant not just participation in politics but primarily the ability to benefit the city in a wide range of potential ways. Thus the education, training, and even inspiration provided in the pederastic relationship released creative forces that led to what has been called the Greek 'miracle.' From around 630 BCE we find the institution of Greek pederasty informing the art and literature to a degree yet to be fully appreciated. Moreover, this influence not only extends to the 'higher' realms of culture, but also can be seen stimulating society at all levels, from the military to athletic games, from philosophy to historiography. An understanding of sexual practices-useful, even essential, to an appreciation of Greek pederasty cannot fully explicate its relationship to these other phenomena; pederasty is found in many societies, and certainly existed before the Greeks. It is time that we move beyond Dover and recover the constructive dynamics of Greek pederasty. PMID- 16338890 TI - The dispersion of pederasty and the athletic revolution in sixth-century BC Greece. AB - Dorian Crete and Thebes are conventionally seen by ancient sources as the originators of pederasty; modern historians see support for this view in Dorian male-centered militarism and sexual segregation in upbringing. Here athletic culture, including training, nudism, and competition, is argued to be a chief 'trigger' for the emergence of pederasty in Sparta and its relatively rapid spread to other Greek states in the seventh to sixth centuries BC. Athletic nudity, in particular, was not a device to enforce civic egalitarianism, as some have argued, but is a persistently erotic incentive that reinforces hegemonic maleness and advertises the individual's virtuous exercise of restraint. In particular, Sparta is found to be the likely source of generalized athletic nudity combined with open pederasty in the early seventh century BC. Nudism in Greek art is erotically charged and not, as others argue, simply a gender marker in the seventh century. Generalized athletic nudity spread to other Greek states emulating the successful Spartan model by the 'athletic revolution' of the early sixth century. With athletic nudity, open pederasty, again following Sparta, was fostered. PMID- 16338891 TI - Glukus himeros: pederastic influence on the myth of Ganymede. AB - Pederastic influence on the myth of Ganymede enables it to evolve, in a continuous line of development easily traced in the history of Greek literature from Homer to Plato, into a homoerotic emblem of the spiritual union of the human and divine. Continuity in this history is marked by the thematic use of the Homeric phrase gammalambdaupsilonkappauvarsigma 'imicroepsilonrhoomicronvarsigma (glukus himeros, sweet longing) to describe sexual desire in association with the Ganymede myth in the Hymn to Aphrodite, Pindar and Plato. PMID- 16338892 TI - Pindar's Tenth Olympian and athlete-trainer pederasty. AB - The comparison of the adolescent boxer Hagesidamus and his trainer Ilas to Patroclus and Achilles in Pindar's Olympian 10.16-21 and the subsequent comparison of Hagesidamus to Ganymede in Olympian 10.99-105 suggest that the relationship was in some sense pederastic, particularly in the wake of Aeschylus' treatment of Achilles and Patroclus in these terms in Myrmidons. This possibility motivates a broader examination of the evidence for such relationships in fifth century Greece. There is no doubt that the palaestra was a central locus for the formation of pederastic liaisons and that athletic nudity was integral to the esthetic construction of adolescent beauty. There is also no doubt that the trainer's position afforded him regular intimacy and close physical contact with boys; several Hellenistic texts take for granted the erotic opportunities connected with the position. The Solonian law presuming to protect pupils from such relationships, attested in Aeschines, was probably a late fifth-century development in reaction to their common occurrence in earlier generations. Evidence also exists for lovers acting as financial backers to boy athletes or as informal trainers. Some of the most intriguing evidence for the conflation of the trainer's and lover's roles can be found in red-figure vase painting of the late sixth and fifth centuries. PMID- 16338893 TI - Boeotian swine: homosexuality in Boeotia. AB - This article shows that the accounts in our ancient sources regarding Boeotian attitudes towards homosexuality, namely that the Boeotians were different from other Greeks in that they enjoyed great freedom in this respect and seemingly everything was permissible to them, present a distorted picture of the homosexual practices in this region. In fact, vase paintings with homosexual iconography dating from the sixth century BC reveal marked similarities with Attic and Corinthian pottery ware of the same period. The view that the Boeotians conducted themselves in an 'uncivilized' manner in their homosexual relations is therefore better understood as an attempt by other Greeks to distinguish themselves from the 'boorish' Boeotians and to justify their own aversion to this form of erotic love. PMID- 16338894 TI - "Sleeping in the bosom of a tender companion": homoerotic attachments in Sappho. AB - This paper reexamines the ancient evidence to see what light it sheds on homoeroticism in Sappho. From the Hellenistic period on there are derogatory references to her homosexuality-and also denials that she was involved in same sex relationships. From the late archaic period on there are hints that women from Lesbos had a reputation for being sexually adventurous. Yet there is a discontinuity between these quips about Sappho and/or Lesbianism, and her own poetry, which is intense, sometimes voluptuous, but really not very carnal. Sappho's oeuvre is so fragmentary that the evidence it offers is tentative at best. Nevertheless, if her homoerotic poetry is at all autobiographical it reflects a circle of mainly adolescent girls or very young women around a somewhat older and more authoritative Sappho. Passionate attachments exist between members of this group as well as between individual girls and Sappho. Although many modern scholars believe Sappho's relationships were egalitarian and same-age, the collective evidence of her own poetry together with the ancient testimonia and commentaries does not support that inference. PMID- 16338895 TI - Some myths and anomalies in the study of Roman sexuality. AB - This paper seeks to dispel several myths prevalent in the scholarship on Roman sexuality: that a freed slave was still obligated to serve his former master's sexual demands (I.A.), that the cinaedus cannot be the same as the modern male homosexual because the cinaedus was thought capable of performing cunnilinctus (I.B.), that exoleti were male prostitutes (I.C.), that the Romans were implacably hostile to lesbianism and that they constructed the lesbian as a phallic monstrosity (II.). It also draws attention to some neglected, unfamiliar, or misinterpreted evidence-anomalous on the current understanding of Roman sexuality, where women, boys, and lower-class men are supposed to have equal standing as potential passive sexual partners for adult men-for adult men whose sexual partners are exclusively male, and either active or passive: exoleti as active partners (I.C.), a puer delicatus who is prized for a masculine appearance rather than a feminine one (I.D.), and the Warren Cup, which glorifies a world of exclusively male-male sexuality (I.E.). PMID- 16338896 TI - Representations of the cinaedus in Roman art: Evidence of "gay" subculture? AB - Whereas analysis of ancient Roman texts reveals signs of a possible homosexual subculture, their interpretation is difficult. This article analyzes the content and context of visual representations of male-male intercourse, including wall paintings at Pompeii, a silver cup, and an engraved agate gemstone. Whether presenting negative stereotypes (Tavern of Salvius, Pompeii; Suburban Baths, Pompeii), or positive ones (Warren Cup, British Museum; Leiden gemstone), these representations reveal the presence of well-developed social attitudes toward the practice ofmale-male sex and the practitioners themselves. PMID- 16338897 TI - The originality of Tibullus' Marathus elegies. AB - As far we can judge from the extant literature, Tibullus' three Marathus elegies are among the most sophisticated poetry of male same-sex desire and love composed in the ancient Greco-Roman world. These poems belong to a long and well established tradition of male homoerotic poetry that goes back to the Greeks of the Archaic Age and was given new impetus centuries later in Roman literature. In this tradition, Tibullus' Marathus elegies stand out for their qualities of irony, dramatic engagement, and psychological finesse. PMID- 16338898 TI - On kissing and sighing: Renaissance homoerotic love from Ficino's De Amore and Sopra Lo Amore to Cesare Trevisani's L'impresa (1569). AB - This essay investigates the homoerotic connotations present in the so-called treatises on love, a popular philosophical and literary genre of the Italian Renaissance. The referential text of this sixteenth- century genre is Marsilio Ficino's De amore (1484), a deeply innovative interpretation of Plato's Symposium. Focusing on the initial section of Ficino's text, Maggi highlights some important structural differences between the De amore and the Symposium. Moreover, by comparing Ficino's Latin text with his own subsequent Italian translation (Sopra lo amore, 1544), Maggi examines how Ficino interprets some key terms such as appearance and splendor. The second part of the essay studies Cesare Trevisani's L'impresa (1569), a later treatise on love with an explicit homoerotic foundation. PMID- 16338899 TI - Light in hellas: how German classical philology engendered gay scholarship. AB - Beginning in the latter part of the eighteenth century, German classical philology acquired a hegemonic status that made it the envy of scholars in other nations. Among the tasks embraced by this great endeavor was the study of what is known of same-sex behavior in ancient Greece. Remarkably, the German philologists chose to present their findings straightforwardly in modern German, accessible to every educated reader. The deposit of this inquiry is the basis of our contemporary knowledge of ancient Greek homosexuality. Moreover, by providing models of homosexual behavior that were more positive than those prevalent in Europe at the time, the research fostered the emergence of the German Gay Movement in 1897. PMID- 16338900 TI - Hellenism and homoeroticism in Shelley and his circle. AB - This paper discusses two leading English Romantic poets- Percy Bysshe Shelley and George Gordon, Lord Byron-and three of their friends, who lived close together in Italy during the first half of 1822. Despite the censorious efforts of family, friends and biographers, ample evidence survives to establish the importance of male love in their lives and works. They were ardent hellenists, whose reference point for male love was the homoerotic ethos of Ancient Greece. PMID- 16338901 TI - The Greek mirror: the Uranians and their use of Greece. AB - The Uranians comprised a loosely knit group of British and American homosexual poets writing between approximately 1880 and 1930, sharing a number of basic cultural and literary assumptions derived on one hand from Walter Pater, and on the other from Walt Whitman. Although they used Oriental, Christian and other motifs, one of the major elements many shared was a use of various allusions and themes from ancient Greece, including paganism, male companionship or intimate friendship (which was not defined in terms of sameness), and democracy and a natural aristocracy of virtue, which they applied to the concerns of their own society and era. The model of male relationships which they advocated (and in at least some cases practiced) was almost uniformly asymmetrical, either by age or class, or both. In addition to their poetry, various theoretical writings by members of the group are also involved in the discussion, and this article argues that these historical/ literary allusions and themes should not be understood as means of evasion which allowed them to write of tabooed subjects safely, but as part of a consciously adopted artistic/cultural strategy for homosexual emancipation. It also suggests that their arguments should be reexamined as a corrective to the present egalitarian model of homosexuality. PMID- 16338902 TI - Eros underground: Greece and Rome in gay print culture, 1953-65. AB - This essay surveys the building of intellectual community through print culture in the nascent gay movement in the United States and in Europe in the mid twentieth century. Amateur historians, especially Jim Kepner and W. Dorr Legg of ONE, used Greece and Rome as models on which to base claims for gay rights. Ancient history figured in ONE's educational enterprises, including articles in the magazine ONE, the ONE Institute, and Homophile Studies. The magazine writers and their readership faced problems in the accessibility of knowledge, which the increasing circulation of the magazines corrected, to a degree. Biases surviving from the Victorian period caused the popular idea of ancient homophile culture to favor Greece over Rome, and made Greek a code word. Antiquity also played a large, though decreasing, role in formations of homoerotic fantasy during this period. PMID- 16338906 TI - Purinergic receptors mediate two distinct glutamate release pathways in hippocampal astrocytes. AB - The purinergic P2X(7) receptor (P2X(7)R) can mediate glutamate release from cultured astrocytes. Using patch clamp recordings, we investigated whether P2X(7)Rs have the same action in hippocampal astrocytes in situ. We found that 2- and 3-O-(4-benzoylbenzoyl)ATP (BzATP), a potent, although unselective P2X(7)R agonist, triggers two different glutamate-mediated responses in CA1 pyramidal neurons; they are transient inward currents, which have the kinetic and pharmacological properties of previously described slow inward currents (SICs) due to Ca(2+)-dependent glutamate release from astrocytes, and a sustained tonic current. Although SICs were unaffected by P2X(7)Rs antagonists, the tonic current was inhibited, was amplified in low extracellular Ca(2+), and was insensitive to glutamate transporter and hemichannel inhibitors. BzATP triggered in astrocytes a large depolarization that was inhibited by P2X(7)R antagonists and amplified in low Ca(2+). In low Ca(2+) BzATP also induced lucifer yellow uptake into a subpopulation of astrocytes and CA3 neurons. Our results demonstrate that purinergic receptors other than the P2X(7)R mediate glutamate release that evokes SICs, whereas activation of a receptor that has features similar to the P2X(7)R, mediates a sustained glutamate efflux that generates a tonic current in CA1 neurons. This sustained glutamate efflux, which is potentiated under non physiological conditions, may have important pathological actions in the brain. PMID- 16338907 TI - Interaction of contractile activity and training history on mRNA abundance in skeletal muscle from trained athletes. AB - Skeletal muscle displays enormous plasticity to respond to contractile activity with muscle from strength- (ST) and endurance-trained (ET) athletes representing diverse states of the adaptation continuum. Training adaptation can be viewed as the accumulation of specific proteins. Hence, the altered gene expression that allows for changes in protein concentration is of major importance for any training adaptation. Accordingly, the aim of the present study was to quantify acute subcellular responses in muscle to habitual and unfamiliar exercise. After 24-h diet/exercise control, 13 male subjects (7 ST and 6 ET) performed a random order of either resistance (8 x 5 maximal leg extensions) or endurance exercise (1 h of cycling at 70% peak O2 uptake). Muscle biopsies were taken from vastus lateralis at rest and 3 h after exercise. Gene expression was analyzed using real time PCR with changes normalized relative to preexercise values. After cycling exercise, peroxisome proliferator-activated receptor-gamma coactivator-1alpha (ET approximately 8.5-fold, ST approximately 10-fold, P < 0.001), pyruvate dehydrogenase kinase-4 (PDK-4; ET approximately 26-fold, ST approximately 39 fold), vascular endothelial growth factor (VEGF; ET approximately 4.5-fold, ST approximately 4-fold), and muscle atrophy F-box protein (MAFbx) (ET approximately 2-fold, ST approximately 0.4-fold) mRNA increased in both groups, whereas MyoD (approximately 3-fold), myogenin (approximately 0.9-fold), and myostatin (approximately 2-fold) mRNA increased in ET but not in ST (P < 0.05). After resistance exercise PDK-4 (approximately 7-fold, P < 0.01) and MyoD (approximately 0.7-fold) increased, whereas MAFbx (approximately 0.7-fold) and myostatin (approximately 0.6-fold) decreased in ET but not in ST. We conclude that prior training history can modify the acute gene responses in skeletal muscle to subsequent exercise. PMID- 16338908 TI - Increased circulating levels of matrix metalloproteinase-2 and -9 in women with the polycystic ovary syndrome. AB - INTRODUCTION: Matrix metalloproteinases (MMPs) have been implicated in various pathological processes including inflammatory response, cardiovascular disease, and recently also in ovarian dysfunction. Polycystic ovary syndrome (PCOS) is the most common endocrinopathy in women of reproductive age and is characterized by chronic anovulation, insulin resistance, and increased prevalence of cardiovascular risk factors. Circulating levels of MMPs and their tissue inhibitors (TIMPs) so far have not been assessed in the PCOS. MATERIALS AND METHODS: Serum levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured in 23 women with PCOS [age (mean +/- sd), 30.5 +/- 6.7 yr; body mass index, 35.8 +/- 7.5 kg/m2] and 22 healthy, regularly menstruating women (age, 29.4 +/- 5.6; body mass index, 31.7 +/- 9.2 kg/m2). RESULTS: Women with PCOS had significantly higher concentrations of MMP-2 (999.8 +/- 155 vs. 521.8 +/- 242 ng/ml; P < 0.001), MMP-9 (592.4 +/- 279 vs. 345 +/- 309; P = 0.007), and TIMP-1 levels (823.8 +/- 145 vs. 692 +/- 210 ng/ml; P = 0.02) than control healthy women. There was no difference in TIMP-2 levels (47.3 +/- 30 vs. 44.4 +/- 39.7 ng/ml; P = 0.21) between women with PCOS and controls. CONCLUSIONS: Obese women with PCOS have elevated serum concentrations of MMP-2 and -9. It might be hypothesized that elevated MMP concentrations may be related to increased cardiovascular risk in PCOS and/or menstrual irregularities associated with this syndrome. PMID- 16338909 TI - Doing things better vs doing better things. PMID- 16338910 TI - Misaligned incentives in America's health: who's minding the store? PMID- 16338911 TI - Time spent in face-to-face patient care and work outside the examination room. AB - PURPOSE: Contrary to physicians' concerns that face-to-face patient time is decreasing, data from the National Ambulatory Medical Care Survey (NAMCS) indicate that between 1988 and 1998, durations of primary care outpatient visits have increased. This study documented how physicians spend time during the workday, including time outside the examination room, and compared observed face to-face patient care time with that reported in NAMCS. METHODS: Using time-motion study techniques, for each of 11 physicians, 2 patient care days were randomly selected and documented by direct observation. Physician time spent on face-to face patient care and 54 activities outside the examination room were documented. Data represent 12,180 minutes of work and 611 outpatient visits. RESULTS: The average workday duration was 8.6 hours, and face-to-face patient care accounted for 55% of the day. Work outside the examination room relevant to a patient currently being seen averaged 14% of the day. Work related to a patient not physically present accounted for one fifth (23%) of the workday. The combination of face-to-face time and time spent on visit-specific work outside the examination room assessed by direct observation was significantly less than the 2003 NAMCS estimate of visit duration assessed by physician report (13.3 vs 18.7 minutes, P <.001). CONCLUSIONS: Nearly one half of a primary care physician's workday is spent on activities outside the examination room, predominately focused on follow-up and documentation of care for patients not physically present. National estimates of visit duration overestimate the combination of face-to-face time and time spent on visit-specific work outside the examination room by 41%. PMID- 16338912 TI - Physician activities during time out of the examination room. AB - PURPOSE: Comprehensive medical care requires direct physician-patient contact, other office-based medical activities, and medical care outside of the office. This study was a systematic investigation of family physician office-based activities outside of the examination room. METHODS: In the summer of 2000, 6 medical students directly observed and recorded the office-based activities of 27 northeastern Ohio community-based family physicians during 1 practice day. A checklist was used to record physician activity every 20 seconds outside of the examination room. Observation excluded medical care provided at other sites. Physicians were also asked to estimate how they spent their time on average and on the observed day. RESULTS: The average office day was 8 hours 8 minutes. On average, 20.1 patients were seen and physicians spent 17.5 minutes per patient in direct contact time. Office-based time outside of the examination room averaged 3 hours 8 minutes or 39% of the office practice day; 61% of that time was spent in activities related to medical care. Charting (32.9 minutes per day) and dictating (23.4 minutes per day) were the most common medical activities. Physicians overestimated the time they spent in direct patient care and medical activities. None of the participating practices had electronic medical records. CONCLUSIONS: If office-based, medically related activities were averaged over the number of patients seen in the office that day, the average office visit time per patient would increase by 7 minutes (40%). Care delivery extends beyond direct patient contact. Models of health care delivery need to recognize this component of care. PMID- 16338913 TI - Quality of work life of independent vs employed family physicians in Wisconsin: a WReN study. AB - PURPOSE: Family physicians in Wisconsin who are mainly employed by large health care organizations have voiced concerns regarding the quality of their work lives. We explored the quality of work life and its relationship to employment by health care organizations. METHODS: We conducted a cross-sectional survey of the 1,482 active members of the Wisconsin Academy of Family Physicians in 2000. RESULTS: A 47% overall response rate was obtained, and 584 respondents could be identified as independent or employed by a health care organization. There were no differences in age or sex between the 2 groups. The independent physicians worked longer hours, were in smaller work groups, and had been in practice longer and in their current practice longer than the employed physicians. Independent physicians reported better working relationships, more satisfaction with family time, more influence over management decisions, better satisfaction with being a physician, better perceived quality of the care they provided, greater ability to achieve professional goals, and lesser intention to leave the practice. CONCLUSIONS: Independent physicians have significantly more positive ratings of several aspects of the quality of their work life compared with physicians employed by health care organizations. Health care organizations need to address these issues if they are to have a satisfied and stable workforce. PMID- 16338914 TI - Physicians answer more clinical questions and change clinical decisions more often with synthesized evidence: a randomized trial in primary care. AB - PURPOSE: Clinicians need evidence in a format that rapidly answers their questions. DynaMed is a database of synthesized evidence. We investigated whether primary care clinicians would answer more clinical questions, change clinical decision making, and alter search time using DynaMed in addition to their usual information sources. METHODS: Fifty-two primary care clinicians naive to DynaMed searched for answers to 698 of their own clinical questions using the Internet. On a per-question basis, participants were randomized to have access to DynaMed (A) or not (N) in addition to their usual information sources. Outcomes included proportions of questions answered, proportions of questions with answers that changed clinical decision making, and median search times. The statistical approach of per-participant analyses of clinicians who asked questions in both A and N states was decided before data collection. RESULTS: Among 46 clinicians in per-participant analyses, 23 (50%) answered a greater proportion of questions during A than N, and 13 (28.3%) answered more questions during N than A (P = .05). Finding answers that changed clinical decision making occurred more often during A (25 clinicians, 54.3%) than during N (13 clinicians, 28.3%) (P = .01). Search times did not differ significantly. Overall, participants found answers for 263 (75.8%) of 347 A questions and 250 (71.2%) of 351 N questions. Answers changed clinical decision making for 224 (64.6%) of the A questions and 209 (59.5%) of the N questions. CONCLUSIONS: Using DynaMed, primary care clinicians answered more questions and changed clinical decisions more often, without increasing overall search time. Synthesizing results of systematic evidence surveillance is a feasible method for meeting clinical information needs in primary care. PMID- 16338915 TI - Quick assessment of literacy in primary care: the newest vital sign. AB - PURPOSE: Current health literacy screening instruments for health care settings are either too long for routine use or available only in English. Our objective was to develop a quick and accurate screening test for limited literacy available in English and Spanish. METHODS: We administered candidate items for the new instrument and also the Test of Functional Health Literacy in Adults (TOFHLA) to English-speaking and Spanish-speaking primary care patients. We measured internal consistency with Cronbach's alpha and assessed criterion validity by measuring correlations with TOFHLA scores. Using TOFLHA scores <75 to define limited literacy, we plotted receiver-operating characteristics (ROC) curves and calculated likelihood ratios for cutoff scores on the new instrument. RESULTS: The final instrument, the Newest Vital Sign (NVS), is a nutrition label that is accompanied by 6 questions and requires 3 minutes for administration. It is reliable (Cronbach alpha >0.76 in English and 0.69 in Spanish) and correlates with the TOFHLA. Area under the ROC curve is 0.88 for English and 0.72 for Spanish versions. Patients with more than 4 correct responses are unlikely to have low literacy, whereas fewer than 4 correct answers indicate the possibility of limited literacy. CONCLUSION: NVS is suitable for use as a quick screening test for limited literacy in primary health care settings. PMID- 16338916 TI - Suicidal ideation and risk levels among primary care patients with uncomplicated depression. AB - PURPOSE: We investigated the prevalence, severity, and course of passive and active suicidal ideation occurring in primary care patients with an uncomplicated depressive disorder. METHODS: We studied suicidal ideation experienced by patients recruited in 60 primary care practices participating in a randomized controlled trial of depression management. Risk levels associated with suicidal ideation and plans were determined by a 2-stage procedure using pertinent items of the Patient Health Questionnaire-9, the Hopkins Symptom Checklist-20, and the Cornell structured assessment interview and management algorithm. RESULTS: Of the 761 patients whom physicians judged in need of treatment for a clinical depression, 405 (53%) were experiencing uncomplicated dysthymia, major depression, or both. Among these depressed patients, about 90% had no risk or a low risk of self-harm based on the presence and nature of suicidal ideation; the rest had an intermediate risk. Almost all patients who were initially classified at the no or low risk levels remained at these levels during the subsequent 6 months. The incidence of suicidal ideation at a risk level requiring the physician's immediate attention in this no- or low-risk subgroup was 1.1% at 3 months and 2.6% at 6 months. CONCLUSIONS: Almost all patients with uncomplicated dysthymia, major depression, or both acknowledging suicidal ideation of the minimal risk type when initially assessed maintained this minimal risk status during the subsequent 6 months. PMID- 16338917 TI - Improving detection of suicidal ideation among depressed patients in primary care. AB - PURPOSE: Primary care clinicians have difficulty detecting suicidal patients. This report evaluates the effect of 2 primary care interventions on the detection and subsequent referral or treatment of patients with depression and recent suicidal ideation. METHODS: Adult patients in 12 mixed-payer primary care practices and 9 not-for-profit staff model health maintenance organization (HMO) practices were screened for depression. Matched practices were randomized within plan type to intervention or usual care. The intervention for mixed-payer practices entailed brief training of physicians and office nurses to provide care management. The intervention for HMO practices consisted of guided development of quality improvement teams for depression care. A total of 880 enrolled patients met study criteria for depression, 232 of whom met criteria for recent suicidal ideation. Intervention effects on suicide detection and referral to mental health specialty care were evaluated with mixed-effects multilevel models in intent-to treat analyses. RESULTS: Depressed patients with recent suicidal ideation were detected on 40.7% of index visits in intervention practices, compared with 20.5% in usual care practices (odds ratio = 2.64, 95% confidence interval, 1.45-5.07), with HMO plan type and male sex associated with detection. The interventions had no effect on referral of patients, starting an antidepressant, or suicidal ideation reported at a 6-month follow-up, although power was limited for all 3 analyses. CONCLUSIONS: Primary care interventions to improve depression care can improve detection of recent suicidal ideation. Further work is needed to improve physician response to detection, including referral to specialty care and more aggressive treatment, and to observe the effect on outcomes. PMID- 16338918 TI - How experiencing preventable medical problems changed patients' interactions with primary health care. AB - PURPOSE: We wanted to explore how patients' experiences with preventable problems in primary care have changed their behavioral interactions with the health care system. METHODS: We conducted semistructured interviews with 24 primary care patients, asking them to describe their experiences with self-perceived preventable problems. We analyzed these interviews using the editing method and classified emotional and behavioral responses to experiencing preventable problems. RESULTS: Anger was the most common emotional response, followed by mistrust and resignation. We classified participants' behavioral responses into 4 categories: avoidance (eg, stop going to the doctor), accommodation (eg, learn to deal with delays), anticipation (eg, attend to details, attend to own emotions, acquire knowledge, actively communicate), and advocacy (eg, get a second opinion). CONCLUSIONS: Understanding how patients react to their experiences with preventable problems can assist health care at both the physician-patient and system levels. We propose an association of mistrust with the behaviors of avoidance and advocacy, and suggest that further research explore the potential impact these patient behaviors have on the provision of health care. PMID- 16338920 TI - To care is to coprovide. AB - Although primary care, including family medicine, recognizes different types of clinician-patient interaction, I argue that only interactions characterized by coprovision define care. By coprovision I mean that clinicians and patients each provide the expertise in health care that they have the capacity to contribute in any given situation. I argue that paternalism and consumerism cannot signify care in any real sense. Some implications of this analysis include a reconceptualization of family medicine and its defining attributes; support for features of caring relationships, such as mutual responsiveness and responsibility; and an acknowledgment that clinicians and patients need to be self-regarding as well as other-regarding. In a previous issue of the Annals, I called for a new dictionary for family medicine, one that would redefine attributes of family medicine in ways not exclusively clinician-centric. Specifically, it would acknowledge the role of patients and their informal caregivers as coproviding, not merely consuming, health care. PMID- 16338919 TI - The break-even point: when medical advances are less important than improving the fidelity with which they are delivered. AB - Society invests billions of dollars in the development of new drugs and technologies but comparatively little in the fidelity of health care, that is, improving systems to ensure the delivery of care to all patients in need. Using mathematical arguments and a nomogram, we demonstrate that technological advances must yield dramatic, often unrealistic increases in efficacy to do more good than could be accomplished by improving fidelity. In 2 examples (the development of anti-platelet agents and statins), we show that enhanced efficacy failed to achieve the health gains that would have occurred by delivering older agents to all eligible patients. Society's huge investment in technological innovations that only modestly improve efficacy, by consuming resources needed for improved delivery of care, may cost more lives than it saves. The misalignment of priorities is driven partly by the commercial interests of industry and by the public's appetite for technological breakthroughs, but health outcomes ultimately suffer. Health, economic, and moral arguments make the case for spending less on technological advances and more on improving systems for delivering care. PMID- 16338921 TI - The current status of medical student education in family medicine. PMID- 16338922 TI - Scholarly activity and residency training: seeking strategic partnerships. PMID- 16338923 TI - UK lessons for US primary care. PMID- 16338924 TI - Shaping the future of pay-for-performance programs. PMID- 16338927 TI - Identification of p122RhoGAP (deleted in liver cancer-1) Serine 322 as a substrate for protein kinase B and ribosomal S6 kinase in insulin-stimulated cells. AB - Protein kinase B (PKB or Akt) plays an essential role in the actions of insulin, cytokines, and growth factors, although the substrates for PKB that are relevant to many of its actions require identification. In this study, we have reported the identification of p122RhoGAP, a GTPase-activating protein selective for RhoA and rodent homologue of the tumor suppressor deleted in liver cancer (DLC1) as a novel insulin-stimulated phosphoprotein in primary rat adipocytes. We have demonstrated that Ser-322 is phosphorylated upon insulin stimulation of intact cells and that this site is directly phosphorylated in vitro by PKB and ribosomal S6 kinase, members of the AGC (protein kinases A, G, and C) family of insulin stimulated protein kinases. Furthermore, expression of constitutively active mutants of PKB or mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) stimulates Ser-322 phosphorylation in intact cells, demonstrating that activation of the PKB or MEK pathway is sufficient for Ser-322 phosphorylation in vivo. Indeed, in primary adipocytes, insulin-stimulated Ser 322 phosphorylation was almost exclusively regulated by the phosphatidylinositol 3-kinase/PKB pathway, whereas in immortalized cells, insulin-stimulated phosphorylation was predominantly regulated by the MEK/extracellular signal regulated kinase/ribosomal S6 kinase pathway, with the phosphatidylinositol 3 kinase/PKB pathway playing a minor role. These results demonstrate that p122RhoGAP Ser-322 acts as an integrator of signal transduction in a manner dependent on the cellular context. PMID- 16338928 TI - Protein kinase C negatively regulates Akt activity and modifies UVC-induced apoptosis in mouse keratinocytes. AB - Skin keratinocytes are subject to frequent chemical and physical injury and have developed elaborate cell survival mechanisms to compensate. Among these, the Akt/protein kinase B (PKB) pathway protects keratinocytes from the toxic effects of ultraviolet light (UV). In contrast, the protein kinase C (PKC) family is involved in several keratinocyte death pathways. During an examination of potential interactions among these two pathways, we found that the insulin-like growth factor (IGF-1) activates both the PKC and the Akt signaling pathways in cultured primary mouse keratinocytes as indicated by increased phospho-PKC and phospho-Ser-473-Akt. IGF-1 also selectively induced translocation of PKCdelta and PKCepsilon from soluble to particulate fractions in mouse keratinocytes. Furthermore, the PKC-specific inhibitor, GF109203X, increased IGF-1-induced phospho-Ser-473-Akt and Akt kinase activity and enhanced IGF-1 protection from UVC-induced apoptosis. Selective activation of PKC by 12-O-tetradecanoylphorbol 13-acetate (TPA) reduced phospho-Ser-473-Akt, suggesting that activation of PKC inhibits Akt activity. TPA also attenuated IGF-1 and epidermal growth factor induced phospho-Ser-473-Akt, reduced Akt kinase activity, and blocked IGF-1 protection from UVC-induced apoptosis. The inhibition of Akt activity by TPA was reduced by inhibitors of protein phosphatase 2A, and TPA stimulated the association of phosphatase 2A with Akt. Individual PKC isoforms were overexpressed in cultured keratinocytes by transduction with adenoviral vectors or inhibited with PKC-selective inhibitors. These studies indicated that PKCdelta and PKCepsilon were selectively potent at causing dephosphorylation of Akt and modifying cell survival, whereas PKCalpha enhanced phosphorylation of Akt on Ser 473. Our results suggested that activation of PKCdelta and PKCepsilon provide a negative regulation for Akt phosphorylation and kinase activity in mouse keratinocytes and serve as modulators of cell survival pathways in response to external stimuli. PMID- 16338929 TI - Crystal structure of the C2 domain of class II phosphatidylinositide 3-kinase C2alpha. AB - Phosphatidylinositide (PtdIns) 3-kinase catalyzes the addition of a phosphate group to the 3'-position of phosphatidyl inositol. Accumulated evidence shows that PtdIns 3-kinase can provide a critical signal for cell proliferation, cell survival, membrane trafficking, glucose transport, and membrane ruffling. Mammalian PtdIns 3-kinases are divided into three classes based on structure and substrate specificity. A unique characteristic of class II PtdIns 3-kinases is the presence of both a phox homolog domain and a C2 domain at the C terminus. The biological function of the C2 domain of the class II PtdIns 3-kinases remains to be determined. We have determined the crystal structure of the mCPK-C2 domain, which is the first three-dimensional structural model of a C2 domain of class II PtdIns 3-kinases. Structural studies reveal that the mCPK-C2 domain has a typical anti-parallel beta-sandwich fold. Scrutiny of the surface of this C2 domain has identified three small, shallow sulfate-binding sites. On the basis of the structural features of these sulfate-binding sites, we have studied the lipid binding properties of the mCPK-C2 domain by site-directed mutagenesis. Our results show that this C2 domain binds specifically to PtdIns(3,4)P(2) and PtdIns(4,5)P(2) and that three lysine residues at SBS I site, Lys-1420, Lys-1432, and Lys-1434, are responsible for the phospholipid binding affinity. PMID- 16338930 TI - Chronic wasting disease of elk and deer and Creutzfeldt-Jakob disease: comparative analysis of the scrapie prion protein. AB - Chronic wasting disease (CWD), a transmissible prion disease that affects elk and deer, poses new challenges to animal and human health. Although the transmission of CWD to humans has not been proven, it remains a possibility. If this were to occur, it is important to know whether the "acquired" human prion disease would show a phenotype including the scrapie prion protein (PrP(Sc)) features that differ from those associated with human sporadic prion disease. In this study, we have compared the pathological profiles and PrP(Sc) characteristics in brains of CWD-affected elk and deer with those in subjects with sporadic Creutzfeldt-Jakob disease (CJD), as well as CJD-affected subjects who might have been exposed to CWD, using histopathology, immunohistochemistry, immunoblotting, conformation stability assay, and N-terminal protein sequencing. Spongiform changes and intense PrP(Sc) staining were present in several brain regions of CWD-affected animals. Immunoblotting revealed three proteinase K (PK)-resistant bands in CWD, representing different glycoforms of PrP(Sc). The unglycosylated PK-resistant PrP(Sc) of CWD migrated at 21 kDa with an electrophoretic mobility similar to that of type 1 human PrP(Sc) present in sporadic CJD affecting subjects homozygous for methionine at codon 129 (sCJDMM1). N-terminal sequencing showed that the PK cleavage site of PrP(Sc) in CWD occurred at residues 82 and 78, similar to that of PrP(Sc) in sCJDMM1. Conformation stability assay also showed no significant difference between elk CWD PrP(Sc) and the PrP(Sc) species associated with sCJDMM1. However, there was a major difference in glycoform ratio of PrP(Sc) between CWD and sCJDMM1 affecting both subjects potentially exposed to CWD and non-exposed subjects. Moreover, PrP(Sc) of CWD exhibited a distinct constellation of glycoforms distinguishable from that of sCJDMM1 in two dimensional immunoblots. These findings underline the importance of detailed PrP(Sc) characterization in trying to detect novel forms of acquired prion disease. PMID- 16338931 TI - Targeting prostaglandin E2 receptors as an alternative strategy to block cyclooxygenase-2-dependent extracellular matrix-induced matrix metalloproteinase 9 expression by macrophages. AB - COX-2-dependent prostaglandin (PG) E2 synthesis regulates macrophage MMP expression, which is thought to destabilize atherosclerotic plaques. However, the administration of selective COX-2 inhibitors paradoxically increases the frequency of adverse cardiovascular events potentially through the loss of anti inflammatory prostanoids and/or disturbance in the balance of pro- and anti thrombotic prostanoids. To avoid these collateral effects of COX-2 inhibition, a strategy to identify and block specific prostanoid-receptor interactions may be required. We previously reported that macrophage engagement of vascular extracellular matrix (ECM) triggers proteinase expression through a MAPKerk1/2 dependent increase in COX-2 expression and PGE2 synthesis. Here we demonstrate that elicited macrophages express the PGE2 receptors EP1-4. When plated on ECM, their expression of EP2 and EP4, receptors linked to PGE2-induced activation of adenylyl cyclase, is strongly stimulated. Forskolin and dibutryl cyclic-AMP stimulate macrophage matrix metalloproteinase (MMP)-9 expression in a dose dependent manner. However, an EP2 agonist (butaprost) has no effect on MMP-9 expression, and macrophages from EP2 null mice exhibited enhanced COX-2 and MMP-9 expression when plated on ECM. In contrast, the EP4 agonist (PGE1-OH) stimulated macrophage MMP-9 expression, which was inhibited by the EP4 antagonist ONO-AE3 208. When compared with COX-2 silencing by small interfering RNA or inhibition by celecoxib, the EP4 antagonist was as effective in inhibiting ECM-induced proteinase expression. In addition, ECM-induced MMP-9 expression was blocked in macrophages in which EP4 was silenced by small interfering RNA. Thus, COX-2 dependent ECM-induced proteinase expression is effectively blocked by selective inhibition of EP4, a member of the PGE2 family of receptors. PMID- 16338932 TI - E-cadherin-dependent transcriptional control of apolipoprotein A-IV gene expression in intestinal epithelial cells: a role for the hepatic nuclear factor 4. AB - Cell-matrix and cell-cell adhesion play a central role in the control of cell proliferation, differentiation, and gene expression. Integrins and E-cadherin are the key components involved in these processes in epithelial cells. We recently showed that integrin-dependent adhesion to the extracellular matrix reinforces the formation of E-cadherin-actin complexes inducing the polarization of Caco-2 enterocytes and increases the expression of a marker of enterocyte differentiation, the apolipoprotein A-IV (apoA-IV) gene. By impairing or enhancing E-cadherin-dependent cell adhesion, we demonstrate in the present study its involvement in the transcriptional activation of the apoA-IV gene in Caco-2 cells. This control requires the regulatory sequence that we have previously identified as necessary and sufficient to drive and restrict apoA-IV gene expression in enterocytes in vivo. Furthermore, using chimeric E-cadherin-Fc homophilic ligand-coated surfaces, we show that a direct activation of E-cadherin triggers the transcriptional activation of the apoA-IV promoter. Finally, E cadherin-dependent cell-cell adhesion controls the nuclear abundance of the transcription factor hepatic nuclear factor 4alpha, which is involved in the enterocyte-specific expression of apoA-IV gene. Altogether, our results suggest that E-cadherin controls enterocyte-specific expression of genes, such as the apoA-IV gene, through the control of hepatic nuclear factor 4alpha nuclear abundance. PMID- 16338933 TI - Overexpression of apolipoprotein A-IV enhances lipid secretion in IPEC-1 cells by increasing chylomicron size. AB - Intestinal apolipoprotein A-IV expression is highly regulated by dietary lipid in newborn swine, suggesting a role in lipid absorption. Constitutive overexpression of apoA-IV in newborn swine enterocytes enhances basolateral secretion of triacylglycerol (TG) in TG-rich lipoproteins 4.9-fold (Lu, S., Yao, Y., Meng, S., Cheng, X., and Black, D. D. (2002) J. Biol. Chem. 277, 31929-31937). To investigate the mechanism of this enhancement, IPEC-1 cells were transfected with a tetracycline-regulatable expression system (Tet-On). In cells incubated with oleic acid, a dose response relationship was observed between medium doxycycline concentration and basolateral apoA-IV and TG secretion. Similarly regulated expression of apoA-I did not enhance lipid secretion. The mean diameter of TG rich lipoproteins secreted from doxycycline-treated cells was larger than from untreated cells (87.0 nm versus 53.4 nm). Basolateral apoB secretion decreased. Using the same expression system, full-length human apoA-IV (376 amino acids); a "pig-like" human apoA-IV, lacking the C-terminal EQQQ repeats (361 amino acids); and a "chicken-like" apoA-IV, further truncated to 343 amino acids, were expressed in IPEC-1 cells. With increasing protein secretion, cells expressing the full-length human apoA-IV displayed a 2-fold increase in TG secretion; in sharp contrast, cells expressing the pig-like human apoA-IV displayed a 25-fold increase in TG secretion and a 27-fold increase in lipoprotein diameter. When human apoA-IV was further truncated to yield a chicken-like protein, TG secretion was inhibited. We conclude that overexpression of swine apoA-IV enhances basolateral TG secretion in a dose-dependent manner by increasing the size of secreted lipoproteins. These data suggest that the region in the human apoA-IV protein from residues 344 to 354 is critical to its ability to enhance lipid secretion, perhaps by enabling the packaging of additional core TG into chylomicron particles. The EQQQ-rich region may play an inhibitory or modulatory role in chylomicron packaging in humans. PMID- 16338934 TI - Involvement of myosin Vb in glutamate receptor trafficking. AB - Myosin V motors mediate cargo transport; however, the identity of neuronal molecules transported by these proteins remains unknown. Here we show that myosin Vb is expressed in several neuronal populations and associates with the alpha amino-3-hydroxy-5-methyl-4-isoxazole propionate-type glutamate receptor subunit GluR1. In developing hippocampal neurons, expression of the tail domain of myosin Vb, but not myosin Va, enhanced GluR1 accumulation in the soma and reduced its surface expression. These changes were accompanied by reduced GluR1 clustering and diminished frequency of excitatory but not inhibitory synaptic currents. Similar effects were observed upon expression of full-length myosin Vb lacking a C-terminal region required for binding to the small GTPase Rab11. In contrast, mutant myosin Vb did not change the localization of several other neurotransmitter receptors, including the glutamate receptor subunit NR1. These results reveal a novel mechanism for the transport of a specific glutamate receptor subunit in neurons mediated by a member of the myosin V family. PMID- 16338935 TI - A unique ATP hydrolysis mechanism of single-headed processive myosin, myosin IX. AB - Recent studies have revealed that myosin IX is a single-headed processive myosin, yet it is unclear how myosin IX can achieve the processive movement. Here we studied the mechanism of ATP hydrolysis cycle of actomyosin IXb. We found that myosin IXb has a rate-limiting ATP hydrolysis step unlike other known myosins, thus populating the prehydrolysis intermediate (M.ATP). M.ATP has a high affinity for actin, and, unlike other myosins, the dissociation of M.ATP from actin was extremely slow, thus preventing myosin from dissociating away from actin. The ADP dissociation step was 10-fold faster than the overall ATP hydrolysis cycle rate and thus not rate-limiting. We propose the following model for single-headed processive myosin. Upon the formation of the M.ATP intermediate, the tight binding of actomyosin IX at the interface is weakened. However, the head is kept in close proximity to actin due to the tethering role of loop 2/large unique insertion of myosin IX. There is enough freedom for the myosin head to find the next location of the binding site along with the actin filament before complete dissociation from the filament. After ATP hydrolysis, Pi is quickly released to form a strong actin binding form, and a power stroke takes place. PMID- 16338936 TI - QRS duration and late mortality in unselected post-infarction patients of the revascularization era. AB - AIMS: To assess the association of prolonged QRS duration and late mortality in unselected post-infarction patients of the revascularization era. METHODS AND RESULTS: A total of 1455 survivors of acute myocardial infarction (MI) in sinus rhythm and under 76 years of age were enrolled. Ninety eight percent of the patients received reperfusion/revascularization therapy (90% percutaneous coronary intervention). After revascularization, prolonged QRS duration (>or=120 ms) was present in 87 patients (6.0%). Additional risk factors studied were age (>or=65 years), presence of diabetes mellitus, history of previous MI, mean heart rate (>75 b.p.m.), heart rate variability index (640 nm), 250 micromol photons m(-2) s(-1) in the absence of HCO3-] caused chloroplasts to take up O2 and accumulate H2O2. Concomitant with the O2 uptake, both ascorbate peroxidase (APX) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) lost their activities. However, superoxide dismutase (SOD), monodehydroascorbate radical reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) activities remained unaffected. The extent to which the photosynthetic linear electron flow decreased was small compared with the decline in APX activity. Therefore, the loss of APX activity lowered the electron flux through the WWC, as evidenced by a decrease in relative electron flux through PSII [Phi(PSII)xPFD]. To verify these interpretations, we created a transplastomic tobacco line in which an H2O2-insensitive APX from the red alga, Galdieria partita, was overproduced in the chloroplasts. In intact transplastomic chloroplasts which were illuminated under conditions that limited photosynthesis, neither O2 uptake nor H2O2 accumulation occurred. Furthermore, the electron flux through the WWC and the activity of GAPDH were maintained. The present work is the first report of APX inactivation by endogenous H2O2 in intact chloroplasts. PMID- 16338961 TI - Renal therapy by stem cells: outsource or in-house? PMID- 16338962 TI - Factor H and atypical hemolytic uremic syndrome: mutations in the C-terminus cause structural changes and defective recognition functions. AB - Atypical hemolytic uremic syndrome is a disease that is characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. Mutations in the complement regulator factor H are associated with the inherited form of the disease, and >60% of the mutations are located within the C terminus of factor H. The C-terminus of factor H, represented by short consensus repeat 19 (SCR19) and SCR20, harbors multiple functions; consequently, this study aimed to examine the functional effects of clinically reported mutations in these SCR. Mutant factor H proteins (W1157R, W1183L, V1197A, R1210C, R1215G, and P1226S) were recombinantly expressed and functionally characterized. All six mutant proteins showed severely reduced heparin, C3b, C3d, and endothelial cell binding. By peptide spot analyses, four linear regions that are involved in heparin, C3b, and C3d binding were localized in SCR19 and SCR20. A three-dimensional homology model of the two domains suggests that these four regions form a common binding site across both domains. In addition, this structural model identifies two types of residues: Type A residues are positioned on the SCR surface and are represented by mutants W1157R, W1183L, R1210C, and R1215G; and type B residues are buried within the SCR structure and affect mutations V1197A and P1226S. Mutations of both types of residue result in the same functional defects, namely the reduced binding of factor H to surface-attached C3b molecules and reduced complement regulatory activity at the cell surfaces. The buried type B mutations seem to affect ligand interaction of factor H more severely than the surface exposed mutations. PMID- 16338963 TI - Prostaglandin E2 inhibits its own renal transport by downregulation of organic anion transporters rOAT1 and rOAT3. AB - Prostaglandin E2 (PGE2) is the principal mediator of fever and inflammation. Recently, evidence emerged that during febrile response, PGE2 that is generated in the periphery enters the hypothalamus and contributes to the maintenance of fever. In a rat model of fever generation, peripheral PGE2 is increased, whereas clearance by metabolism of peripheral PGE2 is downregulated. The major route of PGE2 excretion is via the renal proximal tubular organic anion secretory system, where basolateral uptake that is mediated by renal organic anion transporter 1 (rOAT1) and rOAT3 is rate limiting. Therefore, it was hypothesized that PGE2 itself will abolish its excretion by rOAT1 or rOAT3. Fluorescein was used as a prototypic organic anion, and NRK-52E cells from rat served as a proximal tubular model system. PGE2 time-dependently downregulates basolateral organic anion uptake, without affecting cell volume or cell protein, recirculation of counter ions, or proximal tubular transport systems in general. In addition, PGE2 diminishes expression of both rOAT1 and rOAT3. Both organic anion uptake and expression of rOAT1 and rOAT3 are dose-dependently downregulated by PGE2. These findings suggest that during fever or inflammation, renal secretory transport of PGE2 is reduced, contributing to elevated PGE2 levels in blood. These data fit into the hypothetical concept of peripheral PGE2's playing a significant role in fever. The described regulatory mechanism may also be of relevance in chronic inflammatory events. Moreover, the data presented could explain why increased plasma urate levels occur in diseases that go along with increased levels of PGE2. PMID- 16338964 TI - Src activation of NF-kappaB augments IL-1beta-induced nitric oxide production in mesangial cells. AB - NF-kappaB is a critical transcription factor that is involved in glomerulonephritis and inflammatory host responses and a critical transactivator of the inducible nitric oxide (NO) synthase gene in mesangial cells. The Src protein tyrosine kinases (SFK) are involved in several signaling pathways and have been proposed to mediate cytokine activation of NF-kappaB in a few cell types. However, the specific involvement of SFK in IL-1beta induction of NO production has not been clearly established. Accordingly, pharmacologic and molecular tools were used to clarify this issue in cultured murine mesangial cells. The SFK antagonist 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo(3,4 d)pyrimidine (PP2) dramatically inhibited IL-1beta-mediated induction of endogenous NO production as measured by the Griess reaction, as well as the induction of NF-kappaB p50/p65 DNA-binding activity in gel shift assays and the activity of an NF-kappaB-responsive promoter-reporter construct transiently transfected into the cells. Immunoprecipitation and immunoblotting with anti IkappaBalpha and anti-phosphotyrosine antibodies revealed that PP2 also inhibited IL-1beta-stimulated tyrosine phosphorylation of IkappaBalpha, a requisite step in NF-kappaB activation in this signaling cascade. In agreement with the pharmacologic inhibition studies, siRNA directed against c-Src specifically limited c-Src protein expression and inhibited IL-1beta-mediated induction of NF kappaB DNA-binding activity, whereas control siRNA had no effect. Conversely, overexpression of constitutively active c-Src augmented basal and IL-1beta mediated induction of NF-kappaB DNA-binding activity and NO production. Thus, SFK play a key role in IL-1beta-induced NO production in mesangial cells and do so via tyrosine phosphorylation of IkappaBalpha and consequent NF-kappaB activation. PMID- 16338965 TI - Contrary to rat-type, human-type Na,K-ATPase is phosphorylated at the same amino acid by hormones that produce opposite effects on enzyme activity. AB - Renal sodium homeostasis is a major determinant of BP and is regulated by several natriuretic and antinatriuretic hormones. These hormones, acting through intracellular secondary messengers, either activate or inhibit proximal tubule Na,K-ATPase. It was shown previously that phorbol esters and angiotensin II and serotonin induce the phosphorylation of both Ser-11 and Ser-18 of the Na,K-ATPase alpha-subunit. This results in the recruitment of Na,K-ATPase molecules to the plasma membrane and an increased capacity to transport sodium ions. Treatment of the same cells with dopamine leads to phosphorylation of the Na,K-ATPase alpha subunit Ser-18. The subsequent internalization of Na,K-ATPase molecules results in a reduced capacity to transport sodium ions. These effects are observed in cells that express the rat-type Na,K-ATPase. However, the Na,K-ATPase alpha1 subunit of several species, such as human, pig, and mouse, does not have a Ser-18 in their N-terminal region. Therefore, the possibility exists that, in those species, the Na,K-ATPase is not regulated by the hormones that regulate natriuresis. This study presents evidence that in cells that express the human type Na,K-ATPase, dopamine inhibits and phorbol esters activate the Na,K-ATPase mediated transport. These opposite effects are mediated by the phosphorylation of the same amino acid residue, Ser-11 of Na,K-ATPase alpha1, and the presence of alpha1 Ser-18 is not essential for the hormonal regulation of Na,K-ATPase activity in LLCPK1 cells. It was observed that, whereas the regulatory stimulation of Na,K-ATPase is mediated by protein kinase Cbeta, the regulatory inhibition is mediated by protein kinase Czeta. This is similar to what was demonstrated previously in cells that express the rat-type Na,K-ATPase. PMID- 16338966 TI - Adult kidney tubular cell population showing phenotypic plasticity, tubulogenic capacity, and integration capability into developing kidney. AB - Using in vivo bromodeoxyuridine (BrdU) labeling, a tubular cell population (label retaining tubular cells [LRTC]) was identified recently in normal adult kidneys, which contributes actively to the regeneration process of the kidney after injury. Here, these LRTC are characterized in vitro. The LRTC population was isolated from BrdU-treated rat kidney by FACS. Both LRTC and non-LRTC underwent proliferation and maintained an epithelial phenotype in the presence of tubulogenic growth factors such as EGF, TGF-alpha, IGF-I, and hepatocyte growth factor. It is interesting that LRTC also proliferated without epithelial markers expression in the presence of soluble factors derived from an embryonic kidney metanephric mesenchyme cell line. The type of extracellular matrix strongly influenced the phenotype of LRTC. Furthermore, in three-dimensional collagen gel culture, LRTC formed tubule-like or tubulocystic structures in response to growth factors (hepatocyte growth factor and fibroblast growth factor) that are known to induce kidney cell tubulogenesis in vitro and/or participate in renal regeneration in vivo. In contrast, non-LRTC did not form these structures. When transplanted into the metanephric kidney, LRTC but not non-LRTC were integrated into epithelial components of nephron, including the proximal tubular cells and the ureteric bud. They also differentiated into fibroblast-like cells. Collectively, these findings suggest that LRTC are an adult kidney tubular cell population that shows phenotypic plasticity, tubulogenic capacity, and integration capability into the developing kidney. PMID- 16338967 TI - Phenotypically and functionally distinct CD8+ lymphocyte populations in long-term drug-free tolerance and chronic rejection in human kidney graft recipients. AB - A substantial proportion of long-term kidney graft recipients, including those with a stable renal function in the absence of immunosuppressive therapy, present a skewed T cell receptor (TCR) Vbeta chain usage, essentially in the CD8+ subset. This study analyzed in more detail phenotypical and functional alterations of CD8+ lymphocytes in drug-free tolerant patients (DF-Tol) compared with recipients with chronic rejection (CR). Phenotyping revealed a significant increase in central memory and a decrease in effector CD8+ lymphocytes in DF-Tol versus CR. The expression of CD28+ and CD27+ on these effector cells was significantly decreased in CR. These profiles were stable over time and independent of treatment. Functionally, the CD8+CD28- lymphocytes were less sensitive to apoptosis than their CD8+CD28+ counterparts, without differences in polyclonal proliferation. The CD8+CD28- cells did not express GITR and FoxP3 but were characterized by high levels of preformed perforin and granzyme A, pointing toward a cytotoxic rather than a suppressor function. CD8+CD28- lymphocytes did not show antigen-specific degranulation when co-cultured with targets that bear donor HLA class I antigens, suggesting that the cytotoxicity is directed either to other determinants of the graft or to nongraft epitopes. Of interest, CD8+ cells from DF-Tol displayed the same profile as healthy individuals, indicating an increase in CD8+CD28- effector lymphocytes in CR rather than a decrease in DF Tol. CD8+ lymphocytes from stable kidney recipients under conventional maintenance immunosuppression displayed a mixed profile, independent of treatment and time of sampling. Taken collectively, these data show a strong cytotoxicity associated CD8+CD28- signature in CR and suggest a suppression of pathologic cytotoxicity in DF-Tol. Further prospective studies should assess whether serial CD8+ phenotyping may help to identify patients who are at risk for CR when immunosuppression is tapered. PMID- 16338968 TI - Stimulation by caveolin-1 of the hypotonicity-induced release of taurine and ATP at basolateral, but not apical, membrane of Caco-2 cells. AB - Regulatory volume decrease (RVD) is a protective mechanism that allows mammalian cells to restore their volume when exposed to a hypotonic environment. A key component of RVD is the release of K(+), Cl(-), and organic osmolytes, such as taurine, which then drives osmotic water efflux. Previous experiments have indicated that caveolin-1, a coat protein of caveolae microdomains in the plasma membrane, promotes the swelling-induced Cl(-) current (I(Cl,swell)) through volume-regulated anion channels. However, it is not known whether the stimulation by caveolin-1 is restricted to the release of Cl(-) or whether it also affects the swelling-induced release of other components, such as organic osmolytes. To address this problem, we have studied I(Cl,swell) and the hypotonicity-induced release of taurine and ATP in wild-type Caco-2 cells that are caveolin-1 deficient and in stably transfected Caco-2 cells that express caveolin-1. Electrophysiological characterization of wild-type and stably transfected Caco-2 showed that caveolin-1 promoted I(Cl,swell), but not cystic fibrosis transmembrane conductance regulator currents. Furthermore, caveolin-1 expression stimulated the hypotonicity-induced release of taurine and ATP in stably transfected Caco-2 cells grown as a monolayer. Interestingly, the effect of caveolin-1 was polarized because only the release at the basolateral membrane, but not at the apical membrane, was increased. It is therefore concluded that caveolin-1 facilitates the hypotonicity-induced release of Cl(-), taurine, and ATP, and that in polarized epithelial cells, the effect of caveolin-1 is compartmentalized to the basolateral membrane. PMID- 16338969 TI - Activation of MAPKs in thrombin-stimulated ventricular myocytes is dependent on Ca2+-independent PLA2. AB - Thrombin stimulation of isolated rabbit ventricular myocytes activates a membrane associated, Ca(2+)-independent PLA(2) (iPLA(2)) that selectively hydrolyzes plasmalogen phospholipids and results in increased production of arachidonic acid and lysoplasmenylcholine. To determine whether MAPK regulates myocardial iPLA(2) activity, we isolated ventricular myocytes from rabbit heart by collagenase digestion and pretreated them with MAPK inhibitors before stimulating them with thrombin. Pretreatment with PD-98059 to inhibit p42/44 MAPK or SB-203580 to inhibit p38 MAPK had no significant effect on thrombin-stimulated, membrane associated iPLA(2) activity. Thrombin stimulation resulted in significant increases in both p42/44 and p38 MAPK activity after 2 min. Pretreatment with the iPLA(2)-selective inhibitor bromoenol lactone completely inhibited thrombin stimulated MAPK activity, suggesting that activation of MAPKs was dependent on iPLA(2) activation. Ventricular myocyte MAPK activity was increased by incubation of the myocytes with lysoplasmenylcholine, a metabolite produced by iPLA(2) catalyzed membrane plasmalogen phospholipid hydrolysis. Altogether, these data suggest that activation of MAPKs occurs downstream of and is dependent on iPLA(2) activation in thrombin-stimulated rabbit ventricular myocytes. PMID- 16338970 TI - Novel role for alphavbeta5-integrin in retinal adhesion and its diurnal peak. AB - alpha(v)beta(5)-Integrin is the sole integrin receptor at the retinal pigment epithelium (RPE)-photoreceptor interface and promotes RPE phagocytic signaling to the tyrosine kinase Mer tyrosine kinase (MerTK) once a day in response to circadian photoreceptor shedding. Herein we identify a novel role for alpha(v)beta(5)-integrin in permanent RPE-photoreceptor adhesion that is independent of alpha(v)beta(5)'s function in retinal phagocytosis. To compare retinal adhesion of wild-type and beta(5)-integrin(-/-) mice, we mechanically separated RPE and neural retina and quantified RPE protein and pigment retention with the neural retina. Lack of alpha(v)beta(5)-integrin with normal expression of other RPE integrins greatly weakened retinal adhesion in young mice and accelerated its age-dependent decline. Unexpectedly, the strength of wild-type retinal adhesion varied with a diurnal rhythm that peaked 3.5 h after light onset, after the completion of phagocytosis, when integrin signaling to MerTK is minimal. Permanent alpha(v)beta(5) receptor deficiency attenuated the diurnal peak of retinal adhesion in beta(5)-integrin(-/-) mice. These results identify alpha(v)beta(5)-integrin as the first RPE receptor that contributes to retinal adhesion, a vital mechanism for long-term photoreceptor function and viability. Furthermore, they indicate that alpha(v)beta(5) receptors at the same apical plasma membrane domain of RPE cells fulfill two separate functions that are synchronized by different diurnal rhythms. PMID- 16338971 TI - Norepinephrine-induced oxidative stress causes PC-12 cell apoptosis by both endoplasmic reticulum stress and mitochondrial intrinsic pathway: inhibition of phosphatidylinositol 3-kinase survival pathway. AB - Norepinephrine (NE) induces endoplasmic reticulum (ER) unfolded protein response and reduces maturation and translocation of NE transporter to cell membrane via enhanced formation of reactive oxygen species in PC-12 cells. In the present study, we investigated whether ER stress is also implicated in the proapoptotic effect of NE. We found that the apoptotic effect of NE was associated with increased processing of ER-resident pro-caspase-12, cleavage of caspase-9 and -3, and mitochondrial release of cytochrome c. ER stress was evidenced by upregulation of ER chaperone GRP78 and transcription factor CHOP and the translocation of XBP-1 from the ER to the nucleus by NE. NE also reduced phospho Akt (Ser473), indicating suppression of the phosphatidylinositol 3-kinase (PI3 kinase)/Akt survival pathway. Similar results were produced by thapsigargin. NGF, which promotes the PI3-kinase/Akt activity, reduced the effects of NE and thapsigargin on apoptosis and activation of caspase-12 and -3. However, the effects of NE, but not of thapsigargin, were abolished by pretreatment with SOD and catalase. In contrast, the PI3-kinase inhibitors LY-294002 and wortmannin abolished the protective effects of both SOD/catalase and NGF on NE-induced apoptosis. The functional importance of caspase-12 activation was supported by the use of Z-ATAD-FMK, which reduced the NE-induced processing of caspase-12 and cell apoptosis, but the caspase-12, -9, and -3 inhibitors had no effects on the increase in cytosolic cytochrome c produced by NE. In contrast, the release of mitochondrial cytochrome c was abolished by SOD/catalase and NGF. These results indicate that NE induced cell apoptosis by both ER stress and a mitochondrial death pathway and that the effects of NE were mediated via oxidative stress and inhibition of the PI3-kinase/Akt survival pathway. PMID- 16338972 TI - Regulation of epithelial tubule formation by Rho family GTPases. AB - Previous work has established that the integrin signal transduction pathway plays an important role in the regulation of epithelial tubule formation. Furthermore, it has been demonstrated that Rho-kinase, an effector of the Rho signaling pathway, is an important downstream modulator of collagen-mediated renal and mammary epithelial tubule morphogenesis. In the present study, MDCK cells that expressed mutant dominant-negative, constitutively active Rho family GTPases were used to provide further insight into Rho-GTPase signaling and the regulation of epithelial tubule formation. Using collagen gel overlays on MDCK cells as a model system, we observed phosphorylated myosin light chain (pMLC) at the leading edge of migrating lamellipodia. This epithelial remodeling led to the formation of multicellular branching epithelial tubular structures with extensive tight junctions. However, in cells expressing dominant-negative RhoN19, MLC phosphorylation, epithelial remodeling, and tubule formation were inhibited. Instead, only small apical lumens with a solitary tight junctional ring were observed, providing further evidence that Rho signaling through Rho-kinase is important in the regulation of epithelial tubule formation. Because the present model for the Rho signaling pathway proposes that Rac plays a prominent but reciprocal role in cell regulation, experiments were conducted using cells that expressed constitutively active RacV12. When incubated with collagen gels, RacV12 expressing cells formed small apical lumens with simple tight junctions, suggesting that Rac1 signaling also has a prominent role in the regulation of epithelial morphogenesis. Complementary collagen gel overlay experiments with wild-type MDCK cells demonstrated that endogenous Rac1 activation levels decreased over a time course consistent with lamellipodia and tubule formation. Under these conditions, Rac1 was initially localized to the basolateral membrane. However, after epithelial remodeling, activated Rac1 was observed primarily in lamellipodia. These studies support a model in which Rac1 and RhoA are important modulators of epithelial tubule formation. PMID- 16338973 TI - eNOS translocation but not eNOS phosphorylation is dependent on intracellular Ca2+ in human atrial myocardium. AB - In endothelial cells, two ways of endothelial nitric oxide (NO) synthase (eNOS) activation are known: 1) translocation and 2) Akt-dependent phosphorylation of the enzyme at Ser(1177) (Ser(1177) eNOS). We have recently shown that agonist induced Ser(1177) eNOS phosphorylation also occurs in human myocardium (10). In this study, we investigated the Ca(2+) dependency of these two mechanisms in human atrium. Therefore, atrial tissue was obtained from patients who underwent coronary artery bypass operations. In immunohistochemical experiments, the translocated form of eNOS and phosphorylated Ser(1177) eNOS were labeled using specific antibodies. eNOS translocation was measured in the absence and presence of the Ca(2+) chelator BAPTA before and after application of BRL 37344 (BRL), a beta(3)-adrenoceptor agonist that increases eNOS activity (34). In the absence of BAPTA, BRL time dependently increased the staining intensity of translocated eNOS, whereas in the presence of BAPTA, this effect was blunted. In contrast, BRL clearly increased the staining of phosphorylated Ser(1177) eNOS even in the presence of BAPTA. This observation was confirmed using Western blot analysis. Using the NO-sensitive dye diaminofluorescein, we have demonstrated that BRL induced a strong NO release. This effect was completely abolished in the presence of BAPTA but was unaffected by LY-292004, an inhibitor of phosphatidylinositol 3 kinase activity and eNOS phosphorylation. Although Ca(2+) dependent, neither the translocation of eNOS nor NO release was changed by the adenylate cyclase activator forskolin. In conclusion, 1) in human atrial myocardium, BRL-induced eNOS translocation but not Ser(1177) eNOS phosphorylation is dependent on intracellular Ca(2+). 2) In atrial myocardium, eNOS-translocation and not Ser(1177) eNOS phosphorylation is responsible for generating the main amount of NO. 3) Although Ca(2+) dependent, eNOS translocation and NO release could not be mimicked by adenylate cyclase activation as a mediator of beta-adrenergic stimulation. PMID- 16338974 TI - Role of N- and O-glycans in polarized biosynthetic sorting. AB - The maintenance of proper epithelial function requires efficient sorting of newly synthesized and recycling proteins to the apical and basolateral surfaces of differentiated cells. Whereas basolateral protein sorting signals are generally confined to their cytoplasmic regions, apical targeting signals have been identified that localize to luminal, transmembrane, and cytoplasmic aspects of proteins. In the past few years, both N- and O-linked glycans have been identified as apical sorting determinants. Glycan structures are extraordinarily diverse and have tremendous information potential. Moreover, because the oligosaccharides added to a given protein can change depending on cell type and developmental stage, the potential exists for altering sorting pathways by modulation of the expression pattern of enzymes involved in glycan synthesis. In this review, we discuss the evidence for glycan-mediated apical sorting along the biosynthetic pathway and present possible mechanisms by which these common and heterogeneous posttranslational modifications might function as specific sorting signals. PMID- 16338975 TI - Role of tethering factors in secretory membrane traffic. AB - Coiled-coil and multisubunit tethers have emerged as key regulators of membrane traffic and organellar architecture. The restricted subcellular localization of tethers and their ability to interact with Rabs and soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs) suggests that tethers participate in determining the specificity of membrane fusion. An accepted model of tether function considers them molecular "bridges" that link opposing membranes before SNARE pairing. This model has been extended by findings in various experimental systems, suggesting that tethers may have other functions. Recent reports implicate tethers in the assembly of SNARE complexes, cargo selection and transit, cytoskeletal events, and localized attachment of regulatory proteins. A concept of tethers as scaffolding machines that recruit protein components involved in varied cellular responses is emerging. In this model, tethers function as integration switches that simultaneously transmit information to coordinate distinct processes required for membrane traffic. PMID- 16338976 TI - Cytokine stimulation of pregnancy-associated plasma protein A expression in human coronary artery smooth muscle cells: inhibition by resveratrol. AB - Through specific cleavage of proteins that bind and inhibit insulin-like growth factor-I (IGF-I), pregnancy-associated plasma protein-A (PAPP-A) enhances local IGF-I availability, and, consequently, receptor activation. PAPP-A expression is increased in experimental models of vascular injury and in human atherosclerotic plaque; however, little is known about the regulation of PAPP-A gene expression in vascular cells. In this study, we tested the hypothesis that proinflammatory cytokines involved in the vascular injury response stimulate PAPP-A gene expression in human coronary artery smooth muscle cells (hCASMC) in culture. Tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta stimulated PAPP-A gene expression in a time- and dose-dependent manner. The effect of these cytokines appears to be at the level of transcription because actinomycin D completely prevented the induction of PAPP-A gene expression. Accumulation of PAPP-A in cell-conditioned medium paralleled mRNA synthesis, as did proteolytic activity against IGF binding protein-4 (IGFBP-4). Interestingly, pretreatment of hCASMC with resveratrol, a polyphenol found in the skin of grapes and in red wine purported to underlie the "French paradox," inhibited TNF-alpha- and IL-1beta induced PAPP-A expression and, hence, its IGFBP-4 proteolytic activity. Resveratrol had no effect on basal PAPP-A expression and protease activity. Our finding that PAPP-A gene expression in hCASMC is stimulated by TNF-alpha and IL 1beta suggests a mechanism for the regulation of PAPP-A in response to vascular injury that may contribute to the enhanced IGF-I bioactivity in intimal hyperplasia and atherosclerotic plaque development. Our results also suggest that PAPP-A may be a target of the cardiovascular system-protective effects of resveratrol. PMID- 16338977 TI - Protein kinase C mediates erythrocyte "programmed cell death" following glucose depletion. AB - Glucose depletion of erythrocytes leads to activation of Ca2+-permeable cation channels, Ca2+ entry, activation of a Ca2+-sensitive erythrocyte scramblase, and subsequent exposure of phosphatidylserine at the erythrocyte surface. Ca2+ entry into erythrocytes was previously shown to be stimulated by phorbol esters and to be inhibited by staurosporine and chelerythrine and is thus thought to be regulated by protein phosphorylation/dephosphorylation, presumably via protein kinase C (PKC) and the corresponding phosphoserine/threonine phosphatases. The present experiments explored whether PKC could contribute to effects of energy depletion on erythrocyte phosphatidylserine exposure and cell volume. Phosphatidylserine exposure was estimated from annexin binding and cell volume from forward scatter in fluorescence-activated cell sorter analysis. Removal of extracellular glucose led to depletion of cellular ATP, stimulated PKC activity, led to translocation of PKCalpha, enhanced serine phosphorylation of membrane proteins, decreased cell volume, and increased annexin binding, the latter effect being blunted but not abolished in the presence of 1 microM staurosporine or 50 nM calphostin C. The PKC stimulator phorbol-12-myristate-13-acetate (3 microM) and the phosphatase inhibitor okadaic acid (1-10 microM) mimicked the effect of glucose depletion and similarly led to translocation of PKCalpha and enhanced serine phosphorylation, increased annexin binding, and decreased forward scatter, the latter effects being abrogated by PKC inhibitor staurosporine (1 microM). Fluo-3 fluorescence measurements revealed that okadaic acid also enhanced erythrocyte Ca2+ activity. The present observations suggest that protein phosphorylation and dephosphorylation via PKC and the corresponding protein phosphatases contribute to phosphatidylserine exposure and cell shrinkage after energy depletion. PMID- 16338978 TI - A new mechanism for flow-mediated vasoprotection? Focus on "lung endothelial cell proliferation with decreased shear stress is mediated by reactive oxygen species". PMID- 16338979 TI - Comparison of gene expression in cell culture to that in the intact animal: relevance to drugs and environmental toxicants. Focus on "development of a transactivator in hepatoma cells that allows expression of phase I, phase II, and chemical defense genes". PMID- 16338980 TI - ClC-2 channels get new partners. Focus on "association between Hsp90 and the ClC 2 chloride channel upregulates channel function". PMID- 16338981 TI - Leisure participation predicts survival: a population-based study in Finland. AB - The authors study whether leisure participation is an independent predictor of survival over 20 years. Of the nationally representative sample of 8000 adult Finns (Mini-Finland Health Survey), aged >or=30 years, the cohort of 30-59 years (n 5087) was chosen for the Cox proportional survival analyses. The sum score of leisure participation was divided in quartiles (the lowest quartile = scarce = 0 6), two intermediate quartiles = 7-11 and the highest quartile = abundant = 12 21). Adjusted for statistically significant covariates (age, tobacco smoking, alcohol consumption, obesity, self-rated health and diagnosed chronic diseases), and with scarce participation as the reference, the hazard ratios and 95% confidence intervals (CIs) for the risk of death were 0.80, 0.67-0.95 (intermediate) and 0.66, 0.52-0.84 (abundant) for men. The association was insignificant in women with good health. The results show that leisure participation predicts survival in middle-aged Finnish men and its effect is independent of demographic features, of health status and of several other health related factors. The beneficial effect emphasizes the significance of leisure activities for the promotion of men's health. PMID- 16338982 TI - Europe's precious children. PMID- 16338983 TI - Childhood obesity: the case for binding international legislation. PMID- 16338985 TI - Curbing obesity: prevention and treatment. PMID- 16338986 TI - A checklist for curbing childhood obesity. PMID- 16338987 TI - Dying for heroin: the increasing opioid-related mortality in the Republic of Ireland, 1980-1999. AB - BACKGROUND: Over the past 20 years there has been a steady rise in mortality associated with opioid misuse in several Western countries. We aimed to examine trends in opioid-related mortality over a 20-year period in the Republic of Ireland. METHODS: Retrospective analysis of deaths attributed to ICD-9 codes 304.0 (morphine-type dependence) and E850.0 (accidental poisoning by opiates and related narcotics) in the Republic of Ireland between 1980 and 1999. RESULTS: The Republic of Ireland has seen a rapid increase in the number of opioid-related deaths over the 20-year period studied, from 0.01% of total deaths in 1980 to 0.15% in 1999. This is most marked in the younger age groups where, for example, it rose to 23% of 15-19 year old male deaths for 1997. The opioid-related mortality rate in the 15-44 years age range increased by nearly 14 times between 1980-1984 and 1995-1999. Over the whole period, 87% of opioid-related deaths were amongst males. Outside Dublin there has been a considerable increase in opioid related mortality, nearly doubling the percentage of the total from 6% in the 1980s to 11% in the 1990s. CONCLUSIONS: Opioid-related mortality is an increasing problem in the Republic of Ireland, as in other Western countries. Today, young people have a very low mortality rate, and hence this rapid increase may need specific targeted interventions. The spread of opioid-related mortality outside Dublin to rural and other urban areas will have implications for service planning and provision. PMID- 16338988 TI - The D1 dopamine receptor is constitutively phosphorylated by G protein-coupled receptor kinase 4. AB - G protein-coupled receptor (GPCR) kinases (GRKs) phosphorylate agonist-activated GPCRs, initiating their homologous desensitization. In this article, we present data showing that GRK4 constitutively phosphorylates the D1 receptor in the absence of agonist activation. This constitutive phosphorylation is mediated exclusively by the alpha isoform of GRK4; the beta, gamma, and delta isoforms are ineffective in this regard. Mutational analysis reveals that the constitutive phosphorylation mediated by GRK4alpha is restricted to the distal region of the carboxyl terminus of the receptor, specifically to residues Thr428 and Ser431. Phosphorylation of the D1 receptor by GRK4alpha results in a decrease in cAMP accumulation, an increase in receptor internalization, and a decrease in total receptor number--all of which are abolished in a D1 receptor mutant containing T428V and S431A. The increase in internalized D1 receptors induced by GRK4alpha phosphorylation is due to enhanced receptor internalization rather than retarded trafficking of newly synthesized receptors to the cell surface. The constitutive phosphorylation of the D1 receptor by GRK4alpha does not alter agonist-induced desensitization of the receptor because dopamine pretreatment produced a similar decrease in cAMP accumulation in control cells versus cells expressing GRK4alpha. These observations shift the attenuation of D1 receptor signaling from a purely agonist-driven process to one that is additionally modulated by the complement of kinases that are coexpressed in the same cell. Furthermore, our data provide direct evidence that, in contrast to current dogma, GRKs can (at least in some instances) constitutively phosphorylate GPCRs in the absence of agonist activation resulting in constitutive desensitization. PMID- 16338989 TI - Phosphorylation-dependent desensitization by anandamide of vanilloid receptor-1 (TRPV1) function in rat skeletal muscle arterioles and in Chinese hamster ovary cells expressing TRPV1. AB - It has been proposed that activation of vanilloid receptor-1 (TRPV1) affects the vasotone of resistance arteries. One of the endogenous activators of TRPV1 is anandamide. The effects of anandamide on TRPV1 responsiveness were tested on isolated, pressurized (80 mm Hg) skeletal muscle (m. gracilis) arterioles (179 +/ 33 microm in diameter). We found that the TRPV1 agonist capsaicin (1 microM) elicited a substantial constriction in isolated arterioles (51 +/- 12%). In contrast, anandamide (0-100 microM) did not affect arteriolar diameter significantly (3 +/- 5%). Isolated vessels were also preincubated with anandamide (30 microM for 20 min). This anandamide pretreatment completely blocked capsaicin induced arteriolar constriction (response decreased to 1 +/- 0.6%), and this inhibition was reversed by a protein phosphatase-2B inhibitor (cyclosporin-A; 100 nM, 5 min) treatment (constriction, 31 +/- 1%). An exogenous TRPV1-expressing cell line [Chinese hamster ovary (CHO)-TRPV1] was used to specifically evaluate TRPV1-mediated effects of anandamide. The efficacy of anandamide in this system, as determined by 45Ca2+ uptake, was 65 +/- 8% of that of capsaicin. Upon treatment of the cells with cyclosporin-A or the protein kinase C activator phorbol 12-myristate 13-acetate (PMA), anandamide was transformed to a full agonist. Anandamide treatment caused an acute desensitization in these cells as measured by intracellular Ca2+ imaging. Application of cyclosporin-A or PMA reversed this desensitization. Our data suggest that anandamide may cause a complete (albeit phosphorylation-dependent) desensitization of TRPV1 in skeletal muscle arterioles and in CHO-TRPV1 cells, which apparently transforms the ligand gated TRPV1 into a phosphorylation-gated channel. This property of anandamide may provide a new therapeutic strategy to manipulate TRPV1 activity. PMID- 16338990 TI - Calcium signalling in human spermatozoa: a specialized 'toolkit' of channels, transporters and stores. AB - Ca(2+) is a ubiquitous intracellular messenger which encodes information by temporal and spatial patterns of concentration. In spermatozoa, several key functions, including acrosome reaction and motility, are regulated by cytoplasmic Ca(2+) concentration. Despite the very small size and apparent structural simplicity of spermatozoa, evidence is accumulating that they possess sophisticated mechanisms for regulation of cytoplasmic Ca(2+) concentration and generation of complex Ca(2+) signals. In this review, we consider the various components of the Ca(2+)-signalling 'toolkit' that have been characterized in somatic cells and summarize the evidence for their presence and activity in spermatozoa. In particular, data accumulated over the last few years show that spermatozoa possess one (and probably two) Ca(2+) stores as well as a range of plasma membrane pumps and channels. Selective regulation of the various components of the 'toolkit' by agonists probably allows spermatozoa to generate localized Ca(2+) signals despite their very small cytoplasmic volume, permitting the discrete and selective activation of cell functions. PMID- 16338991 TI - Interactions between asynchronous release and short-term plasticity in the nucleus accumbens slice. AB - Glutamate synapses in the nucleus accumbens (NAc) display asynchronous release in response to trains of stimulation. However, it is unclear what role this asynchronous release plays in synaptic transmission in this nucleus. This process was studied, specifically looking at the interaction between short-term depression and asynchronous release. These results indicate that synchronous and asynchronous release do not compete for a depleted readily releasable pool of vesicles. PMID- 16338992 TI - Texture signals in whisker vibrations. AB - Rodents excel in making texture judgments by sweeping their whiskers across a surface. Here we aimed to identify the signals present in whisker vibrations that give rise to such fine sensory discriminations. First, we used sensors to capture vibration signals in metal whiskers during active whisking of an artificial system and in natural whiskers during whisking of rats in vivo. Then we developed a classification algorithm that successfully matched the vibration frequency spectra of single trials to the texture that induced it. For artificial whiskers, the algorithm correctly identified one texture of eight alternatives on 40% of trials; for in vivo natural whiskers, the algorithm correctly identified one texture of five alternatives on 80% of trials. Finally, we asked which were the key discriminative features of the vibration spectra. Under both artificial and natural conditions, the combination of two features accounted for most of the information: The modulation power-the power of the part of the whisker movement representing the modulation due to the texture surface-increased with the coarseness of the texture; the modulation centroid-a measure related to the center of gravity within the power spectrum-decreased with the coarseness of the texture. Indeed, restricting the signal to these two parameters led to performance three-fourths as high as the full spectra. Because earlier work showed that modulation power and centroid are directly related to neuronal responses in the whisker pathway, we conclude that the biological system optimally extracts vibration features to permit texture classification. PMID- 16338993 TI - Serotonin modulates the properties of ascending commissural interneurons in the neonatal mouse spinal cord. AB - The interneuron populations that constitute the central pattern generator (CPG) for locomotion in the mammalian spinal cord are not well understood. We studied the properties of a set of commissural interneurons whose axons cross and ascend in the contralateral cord (aCINs) in the neonatal mouse. During N-methyl-D aspartate (NMDA) and 5-HT-induced fictive locomotion, a majority of lumbar (L2) aCINs examined were rhythmically active; most of them fired in phase with the ipsilateral motoneuron pool, but some fired in phase with contralateral motoneurons. 5-HT plays a critical role in enabling the locomotor CPG to function. We found that 5-HT increased the excitability of aCINs by depolarizing the membrane potential, reducing the postspike afterhyperpolarization amplitude, broadening the action potential, and decreasing the action potential threshold. Serotonin had no significant effect on the input resistance and sag amplitude of aCINs. These results support the hypothesis that aCINs play important roles in coordinating left-right movements during fictive locomotion and thus may be component neurons in the locomotor CPG in neonatal mice. PMID- 16338994 TI - Encoding changes in orbitofrontal cortex in reversal-impaired aged rats. AB - Previous work in rats and primates has shown that normal aging can be associated with a decline in cognitive flexibility mediated by prefrontal circuits. For example, aged rats are impaired in rapid reversal learning, which in young rats depends critically on the orbitofrontal cortex. To assess whether aging-related reversal impairments reflect orbitofrontal dysfunction, we identified aged rats with reversal learning deficits and then recorded single units as these rats, along with unimpaired aged cohorts and young control rats, learned and reversed a series of odor discrimination problems. We found that the flexibility of neural correlates in orbitofrontal cortex was markedly diminished in aged rats characterized as reversal-impaired in initial training. In particular, although many cue-selective neurons in young and aged-unimpaired rats reversed odor preference when the odor-outcome associations were reversed, cue-selective neurons in reversal-impaired aged rats did not. In addition, outcome-expectant neurons in aged-impaired rats failed to become active during cue sampling after learning. These altered features of neural encoding could provide a basis for cognitive inflexibility associated with normal aging. PMID- 16338995 TI - Dissociated hysteresis of static ocular counterroll in humans. AB - In stationary head roll positions, the eyes are cyclodivergent. We asked whether this phenomenon can be explained by a static hysteresis that differs between the eyes contra- (CE) and ipsilateral (IE) to head roll. Using a motorized turntable, healthy human subjects (n = 8) were continuously rotated about the earth horizontal naso-occipital axis. Starting from the upright position, a total of three full rotations at a constant velocity (2 degrees/s) were completed (acceleration = 0.05 degrees/s2, velocity plateau reached after 40 s). Subjects directed their gaze on a flashing laser dot straight ahead (switched on 20 ms every 2 s). Binocular three-dimensional eye movements were recorded with dual search coils that were modified (wires exiting inferiorly) to minimize torsional artifacts by the eyelids. A sinusoidal function with a first and second harmonic was fitted to torsional eye position as a function of torsional whole body position at constant turntable velocity. The amplitude and phase of the first harmonic differed significantly between the two eyes (paired t-test: P < 0.05): on average, counterroll amplitude of IE was larger [CE: 6.6 +/- 1.6 degrees (SD); IE: 8.1 +/- 1.7 degrees), whereas CE showed more position lag relative to the turntable (CE: 12.5 +/- 10.7 degrees; IE: 5.1 +/- 8.7 degrees). We conclude that cyclodivergence observed during static ocular counterroll is mainly a result of hysteresis that depends on whether eyes are contra- or ipsilateral to head roll. Static hysteresis also explains the phenomenon of residual torsion, i.e., an incomplete torsional return of the eyes when the first 360 degrees whole body rotation was completed and subjects were back in upright position (extorsion of CE: 2.0 +/- 0.10 degrees; intorsion of IE: 1.4 +/- 0.10 degrees). A computer model that includes asymmetric backlash for each eye can explain dissociated torsional hysteresis during quasi-static binocular counterroll. We hypothesize that ocular torsional hysteresis is introduced at the level of the otolith pathways because the direction-dependent torsional position lag of the eyes is related to the head roll position and not the eye position. PMID- 16338996 TI - Spatial integration of optic flow signals in fly motion-sensitive neurons. AB - Neurons in the fly lobula plate integrate motion signals over large regions of visual space in a directionally selective manner. This study is concerned with the details of this integration process. We used a stimulus consisting of a 4 x 4 lattice of locally moving Gabor patches, in which each patch could take any direction independently. We also presented only one patch at a time or two patches at a time. Across all possible directions of motion, the firing rate response r1+2 to two simultaneously presented patches was well described by r1+2(d1, d2) = G x [r1(d1) + r2(d2)] + S, where r1 and r2 are responses to individual patches moving in directions d1 and d2, and G approximately 0.81, S approximately -23. However, this quasi-linear scaling expression failed to account for three main empirical observations: 1) the directional-tuning curve for one patch is broader in the presence of another patch moving in the neuron's preferred direction (PD); 2) the vertical compression of this curve is greater when the second patch moves in the antipreferred direction (AD) as opposed to PD; 3) the ability of the neuronal response to discriminate the direction of a patch is greater when the other patch is moving in the PD as opposed to AD, where this ability is assessed using both information theory and a standard discriminability index. To account for these departures from the simple scaling model, we used a normalization model very similar to one used for macaque area MT/V5. This model can qualitatively explain all three departures from the scaling equation described above, suggesting that a gain-control normalization network may be at work within the fly lobula plate. PMID- 16338997 TI - Sensitivity to interaural time differences in the dorsal nucleus of the lateral lemniscus of the unanesthetized rabbit: comparison with other structures. AB - Interaural time differences, a cue for azimuthal sound location, are first encoded in the superior olivary complex (SOC), and this information is then conveyed to the dorsal nucleus of the lateral lemniscus (DNLL) and inferior colliculus (IC). The DNLL provides a strong inhibitory input to the IC and may serve to transform the coding of interaural time differences (ITDs) in the IC. Consistent with the projections from the SOC, the DNLL and IC had similar distributions of peak- and trough-type neurons, characteristic delays, and best ITDs. The ITD tuning widths of DNLL neurons were intermediate between those of the SOC and IC. Further sharpening is seen in the auditory thalamus, indicating that sharpening mechanisms are not restricted to the midbrain. The proportion of neurons that phase-locked to the tones delivered to each ear progressively decreased from the SOC to the auditory thalamus. The degree of phase-locking for a large majority of DNLL neurons was too weak to support their involvement in processing monaural inputs to generate a sensitivity to ITDs. The response rates of DNLL neurons were on average approximately 60% greater than in the IC or SOC, indicating that the inhibitory input provided to the IC by the DNLL is robust. PMID- 16338998 TI - Electrophysiological heterogeneity of spinally projecting serotonergic and nonserotonergic neurons in the rostral ventromedial medulla. AB - This study examined the passive membrane and action potential properties of serotonergic and nonserotonergic neurons in the rostral ventromedial medulla (RVM) of the rat using whole cell patch-clamp recording techniques in the slice. Serotonergic neurons were identified by immunoreactivity for tryptophan hydroxylase (TrpH). Spinally projecting neurons were retrogradely labeled with 1' dioactadecyl-3,3,3',3'-tetramethylindocarbodyanine perchlorate (DiI). Three types of neurons were identified within both spinally projecting serotonergic and nonserotonergic populations. Type 1 neurons exhibited irregular or sporadic spontaneous activity interspersed with periods of quiescence. Type 2 neurons were not spontaneously active and were additionally discriminated by a more negative resting membrane potential and a larger-amplitude action potential. Type 3 neurons fired repetitively without pause. Serotonergic neurons had a higher membrane resistance and greater action potential half-width than their nonserotonergic counterparts and rarely exhibited a fast afterhyperpolarization. Serotonergic type 3 neurons also fired more slowly and regularly than nonserotonergic type 3 neurons. Comparison of electrophysiological and immunohistochemical characteristics suggested that the smallest type 3 serotonergic neurons had an increased risk of immunohistochemical "misclassification" due to failure to detect TrpH, possibly due to more complete dialysis of intracellular contents during lengthy recordings. This risk was minimal for type 1 or 2 serotonergic neurons. The three different types of spinally projecting serotonergic neurons also differed markedly in their responsiveness to the mu opioid receptor agonist D-Ala2, NMePhe4, Gly5 ol]enkephalin. These results provide important new electrophysiological and pharmacological evidence for a significant heterogeneity among spinally projecting serotonergic RVM neurons. They may also provide a basis for resolving the controversy concerning the role of serotonergic RVM neurons in opioid analgesia. PMID- 16338999 TI - Endogenous acetylcholine enhances synchronized interneuron activity in rat neocortex. AB - Application of 4-aminopyridine (4-AP) along with EAA) receptor antagonists produces gamma-aminobutyric acid (GABAA) receptor-dependent synchronized activity in interneurons. This results in waves of activity propagating through upper cortical layers. Because interneurons in the neocortex are excited by nicotinic acetylcholine receptor (nAChR) agonists, ACh may influence synchronization of these local neocortical interneuronal networks. To study this possibility, we have used voltage-sensitive dye imaging using the fluorescent dye RH 414 (30 microM) in rat neocortical slices. Recordings were obtained in the presence of 4 AP (100 microM) and the EAA receptor antagonists D-2-amino-5-phosphonvaleric acid (20 microM) and 6-cyano-7-nitro-quinoxaline-2,3-dione (10 microM). In response to intracortical stimulation, localized or propagated activity restricted to upper cortical layers was seen. Bath application of the ACh esterase inhibitor neostigmine (10 microM) and the nAChR agonist 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP; 10 microM) increased the response amplitude, the extent of spread, and the duration of this activity. These changes were seen in 13 of 16 slices tested with neostigmine (10 microM) and 4 of 7 slices tested with DMPP (10 microM). Application of the muscarinic AChR antagonist atropine (1 microM) did not block the enhancement of activity by neostigmine (n = 7). Application of dihydro-beta-erythroidine (10 microM), known, at this concentration, to selectively antagonize alpha4beta2-like nAChRs, blocked the effect of neostigmine (n = 5). The selective alpha7-like nAChR antagonist methyllycaconitine (50 nM) was ineffective (n = 5). These results suggest that activation of alpha4beta2 like nAChRs by endogenously released ACh can enhance synchronized activity in local neocortical inhibitory networks. PMID- 16339002 TI - Spatiotemporal activity patterns during respiratory rhythmogenesis in the rat ventrolateral medulla. AB - One of the most important brain rhythms is that which generates involuntary breathing movements. The lower brain stem contains neural circuitry for respiratory rhythm generation in mammals. To date, microsectioning and selective lesioning studies have revealed anatomical regions necessary for respiratory rhythmogenesis. Although respiratory neurons distributed within these regions can be identified by their firing patterns in different phases of the respiratory cycle, conventional electrophysiology techniques have limited the study of spatial organization within this network. Optical imaging techniques offer the potential for monitoring the spatiotemporal activity of large groups of neurons simultaneously. Using high-speed voltage-sensitive dye imaging and spatial correlation analysis in an arterially perfused in situ preparation of the juvenile rat, we determined the spatial distribution of respiratory neuronal activity in a region of the ventrolateral respiratory group containing the pre Botzinger complex (pBC) during spontaneous eupneic breathing. While distinctly pre- and postinspiratory-related responses were spatially localizable on length scales less than 100 microm, we found the studied area on whole exhibited a spatial mixture of phase-spanning and postinspiratory-related activity. Additionally, optical recordings revealed significant widespread hyperpolarization, suggesting inhibition in the same region during expiration. This finding is consistent with the hypothesis that inhibitory neurons play a crucial role in the inspiration-expiration phase transition in the pBC. To our knowledge this is the first optical imaging of a near fully intact in situ preparation that exhibits both eupneic respiratory activity and functional reflexes. PMID- 16339003 TI - Distal gap junctions and active dendrites can tune network dynamics. AB - Gap junctions allow direct electrical communication between CNS neurons. From theoretical and modeling studies, it is well known that although gap junctions can act to synchronize network output, they can also give rise to many other dynamic patterns including antiphase and other phase-locked states. The particular network pattern that arises depends on cellular, intrinsic properties that affect firing frequencies as well as the strength and location of the gap junctions. Interneurons or GABAergic neurons in hippocampus are diverse in their cellular characteristics and have been shown to have active dendrites. Furthermore, parvalbumin-positive GABAergic neurons, also known as basket cells, can contact one another via gap junctions on their distal dendrites. Using two cell network models, we explore how distal electrical connections affect network output. We build multi-compartment models of hippocampal basket cells using NEURON and endow them with varying amounts of active dendrites. Two-cell networks of these model cells as well as reduced versions are explored. The relationship between intrinsic frequency and the level of active dendrites allows us to define three regions based on what sort of network dynamics occur with distal gap junction coupling. Weak coupling theory is used to predict the delineation of these regions as well as examination of phase response curves and distal dendritic polarization levels. We find that a nonmonotonic dependence of network dynamic characteristics (phase lags) on gap junction conductance occurs. This suggests that distal electrical coupling and active dendrite levels can control how sensitive network dynamics are to gap junction modulation. With the extended geometry, gap junctions located at more distal locations must have larger conductances for pure synchrony to occur. Furthermore, based on simulations with heterogeneous networks, it may be that one requires active dendrites if phase locking is to occur in networks formed with distal gap junctions. PMID- 16339000 TI - Neural correlates of attention and distractibility in the lateral intraparietal area. AB - We examined the activity of neurons in the lateral intraparietal area (LIP) during a task in which we measured attention in the monkey, using an advantage in contrast sensitivity as our definition of attention. The animals planned a memory guided saccade but made or canceled it depending on the orientation of a briefly flashed probe stimulus. We measured the monkeys' contrast sensitivity by varying the contrast of the probe. Both subjects had better thresholds at the goal of the saccade than elsewhere. If a task-irrelevant distractor flashed elsewhere in the visual field, the attentional advantage transiently shifted to that site. The population response in LIP correlated with the allocation of attention; the attentional advantage lay at the location in the visual field whose representation in LIP had the greatest activity when the probe appeared. During a brief period in which there were two equally active regions in LIP, there was no attentional advantage at either location. This time, the crossing point, differed in the two animals, proving a strong correlation between the activity and behavior. The crossing point of each neuron depended on the relationship of three parameters: the visual response to the distractor, the saccade-related delay activity, and the rate of decay of the transient response to the distractor. Thus the time at which attention lingers on a distractor is set by the mechanism underlying these three biophysical properties. Finally, we showed that for a brief time LIP neurons showed a stronger response to signal canceling the planned saccade than to the confirmation signal. PMID- 16339004 TI - Temporal nonlinearity during recovery from sequential inhibition by neurons in the cat primary auditory cortex. AB - Auditory stimuli occur most often in sequences rather than in isolation. It is therefore necessary to understand how responses to sounds occurring in sequences differ from responses to isolated sounds. Cells in primary auditory cortex (AI) respond to a large set of binaural stimuli when presented in isolation. The set of responses to such stimuli presented at one frequency comprises a level response area. A preceding binaural stimulus can reduce the size and magnitude of level response areas of AI cells. The present study focuses on the effects of the time interval between a preceding stimulus and the stimuli of a level response area in pentobarbital-anesthetized cats. After the offset of a preceding stimulus, the ability of AI cells to respond to succeeding stimuli varies dynamically in time. At short interstimulus intervals (ISI), a preceding stimulus can completely inhibit responses to succeeding stimuli. With increasing ISIs, AI cells respond first to binaural stimuli that evoke the largest responses in the control condition, i.e., not preceded by a stimulus. Recovery rate is nonlinear across the level response area; responses to these most-effective stimuli recover to 70% of control on average 187 ms before responses to other stimuli recover to 70% of their control sizes. During the tens to hundreds of milliseconds that a level response area is reduced in size and magnitude, the selectivity of AI cells is increased for stimuli that evoke the largest responses. This increased selectivity results from a temporal nonlinearity in the recovery of the level response area which protects responses to the most effective binaural stimuli. Thus in a sequence of effective stimuli, a given cell will respond selectively to only those stimuli that evoke a strong response when presented alone. PMID- 16339005 TI - Phase-locked responses to pure tones in the inferior colliculus. AB - In the auditory system, some ascending pathways preserve the precise timing information present in a temporal code of frequency. This can be measured by studying responses that are phase-locked to the stimulus waveform. At each stage along a pathway, there is a reduction in the upper frequency limit of the phase locking and an increase in the steady-state latency. In the guinea pig, phase locked responses to pure tones have been described at various levels from auditory nerve to neocortex but not in the inferior colliculus (IC). Therefore we made recordings from 161 single units in guinea pig IC. Of these single units, 68% (110/161) showed phase-locked responses. Cells that phase-locked were mainly located in the central nucleus but also occurred in the dorsal cortex and external nucleus. The upper limiting frequency of phase-locking varied greatly between units (80-1,034 Hz) and between anatomical divisions. The upper limits in the three divisions were central nucleus, >1,000 Hz; dorsal cortex, 700 Hz; external nucleus, 320 Hz. The mean latencies also varied and were central nucleus, 8.2 +/- 2.8 (SD) ms; dorsal cortex, 17.2 ms; external nucleus, 13.3 ms. We conclude that many cells in the central nucleus receive direct inputs from the brain stem, whereas cells in the external and dorsal divisions receive input from other structures that may include the forebrain. PMID- 16339006 TI - Expression and function of variants of slob, slowpoke channel binding protein, in Drosophila. AB - Slob binds to and modulates the Drosophila Slowpoke (dSlo) calcium-activated potassium channel and also recruits the ubiquitous signaling protein 14-3-3 to the channel regulatory complex. RT-PCR reveals the presence of multiple slob transcripts in Drosophila heads. The transcripts are predicted to encode proteins that we call Slob51 (kDa), Slob57, Slob65, and Slob71. Slob51 and Slob65 are splice variants that lack a motif important for the binding of 14-3-3. Previous microarray analyses demonstrated the circadian cycling of slob mRNA, and we show by quantitative PCR that more than one transcript cycles in fly heads. Using in situ hybridization, we observe differences in the expression patterns of the different transcripts. Immunohistochemistry on Drosophila heads reveals Slob71/65 protein to be enriched in the lateral neurons, in contrast to Slob57/51 protein, which is expressed most prominently in the pars intercerebralis neurons and dorsal giant interneurons. Using a heterologous expression system, we show that different Slobs bind to different extents to dSlo and 14-3-3. These data reveal an unexpected diversity of the dSlo/Slob/14-3-3 dynamic regulatory complex. PMID- 16339007 TI - Morphine directly inhibits nociceptors in inflamed skin. AB - Peripherally delivered opiates attenuate mechanical and thermal hyperalgesia in experimental models of inflammation, suggesting that activation of peripheral opioid receptors decreases the excitability of nociceptors in inflamed tissues. The current study examines the effects of peripheral morphine sulfate on response properties of sensory neurons in healthy and inflamed skin. Afferent units (185) were isolated from tibial nerve of rats using an in vitro glabrous skin-nerve teased-fiber preparation. Of these, 107 units were from normal healthy skin, and 78 were from inflamed skin 18 h after intraplantar injection of complete Freund's adjuvant. As a population, C-fiber units innervating inflamed skin exhibited properties characteristic of sensitization when compared with units innervating healthy control skin. Mechanical thresholds were lowered, responses to noxious mechanical and thermal stimuli were elevated, a greater proportion of units was spontaneously active, and the average rate of spontaneous discharge was higher. Response properties in other conduction velocity groups remained unchanged. Fifty eight percent of C and C/Adelta nociceptors innervating inflamed skin were opiate sensitive, and their excitability was attenuated by direct application of morphine to their receptive fields. All morphine-sensitive units were nociceptors from inflamed skin with conduction velocities <1.3 m/s. Morphine effects were concentration-dependent and naloxone-sensitive, indicating that the effects were receptor-mediated. These findings provide direct evidence that morphine acts through peripheral opioid receptors to inhibit the activity of cutaneous nociceptors under conditions of inflammation. PMID- 16339008 TI - Eye movements of the murine P/Q calcium channel mutant tottering, and the impact of aging. AB - Mice carrying mutations of the gene encoding the ion pore of the P/Q calcium channel (Cacna1a) are an instance in which cerebellar dysfunction may be attributable to altered electrophysiology and thus provide an opportunity to study how neuronal intrinsic properties dictate signal processing in the ocular motor system. P/Q channel mutations can engender multiple effects at the single neuron, circuit, and behavioral levels; correlating physiological and behavioral abnormalities in multiple allelic strains will ultimately facilitate determining which alterations of physiology are responsible for specific behavioral aberrations. We used videooculography to quantify ocular motor behavior in tottering mutants aged 3 mo to 2 yr and compared their performance to data previously obtained in the allelic mutant rocker and C57BL/6 controls. Tottering mutants shared numerous abnormalities with rocker, including upward deviation of the eyes at rest, increased vestibuloocular reflex (VOR) phase lead at low stimulus frequencies, reduced VOR gain at high stimulus frequencies, reduced gain of the horizontal and vertical optokinetic reflex, reduced time constants of the neural integrator, and reduced plasticity of the VOR as assessed in a cross-axis training paradigm. Unlike rocker, young tottering mutants exhibited normal peak velocities of nystagmus fast phases, arguing against a role for neuromuscular transmission defects in the attenuation of compensatory eye movements. Tottering also differed by exhibiting directional asymmetries of the gains of optokinetic reflexes. The data suggest at least four pathophysiological mechanisms (two congenital and two acquired) are required to explain the ocular motor deficits in the two Cacna1a mutant strains. PMID- 16339009 TI - Comparable GABAergic mechanisms of hippocampal seizurelike activity in posttetanic and low-Mg2+ conditions. AB - It is known that GABA is a major inhibitory neurotransmitter in mature mammalian brains, but the effect of this substance is sometimes converted into depolarizing or even excitatory when the postsynaptic Cl- concentration becomes high. Recently we have shown that seizurelike afterdischarge induced by tetanic stimulation in normal extracellular fluid (posttetanic afterdischarge) is mediated through GABAergic excitation in mature hippocampal CA1 pyramidal cells. In this study, we examined the possible contribution of similar depolarizing/excitatory GABAergic input to the CA1 pyramidal cells to the seizurelike afterdischarge induced in a low extracellular Mg2+ condition, another experimental model of epileptic seizure activity (low-Mg2+ afterdischarge). Perfusion of the GABAA antagonist bicuculline abolished the low-Mg2+ afterdischarge, but not the interictal-like activity, in most cases. Each oscillatory response during the low-Mg2+ afterdischarge was dependent on Cl- conductance and contained an F- -insensitive depolarizing component in the pyramidal cells, thus indicating that the afterdischarge response may be mediated through both GABAergic and nonGABAergic transmissions. In addition, local GABA application to the recorded cells revealed that GABA responses were indeed depolarizing during the low-Mg2+ afterdischarge. Furthermore, the GABAergic interneurons located in the strata pyramidale and oriens fired in oscillatory cycles more actively than those in other layers of the CA1 region. These results suggest that the depolarizing GABAergic input may facilitate oscillatory synchronization among the hippocampal CA1 pyramidal cells during the low-Mg2+ afterdischarge in a manner similar to the expression of the posttetanic afterdischarge. PMID- 16339010 TI - Spirometry for chronic obstructive pulmonary disease case finding in primary care? PMID- 16339011 TI - Environmental health and the AJRCCM. PMID- 16339012 TI - Combined pulmonary fibrosis and emphysema: an experimental and clinically relevant phenotype. PMID- 16339013 TI - Interactions of sepsis, organ dysfunction, and outcome of critically ill children. PMID- 16339014 TI - Study of infliximab treatment in smokers with COPD: what next? PMID- 16339015 TI - Microbial origin of excess methane in glacial ice and implications for life on Mars. AB - Methane trapped in the 3,053-m-deep Greenland Ice Sheet Project 2 ice core provides an important record of millennial-scale climate change over the last 110,000 yr. However, at several depths in the lowest 90 m of the ice core, the methane concentration is up to an order of magnitude higher than at other depths. At those depths we have discovered methanogenic archaea, the in situ metabolism of which accounts for the excess methane. The total concentration of all types of microbes we measured with direct counts of Syto-23-stained cells tracks the excess of methanogens that we identified by their F420 autofluorescence and provides independent evidence for anomalous layers. The metabolic rate we estimated for microbes at those depths is consistent with the Arrhenius relation for rates found earlier for microbes imprisoned in rock, sediment, and ice. It is roughly the same as the rate of spontaneous macromolecular damage inferred from laboratory data, suggesting that microbes imprisoned in ice expend metabolic energy mainly to repair damage to DNA and amino acids rather than to grow. Equating the loss rate of methane recently discovered in the Martian atmosphere to the production rate by possible methanogens, we estimate that a possible Martian habitat would be at a temperature of approximately 0 degrees C and that the concentration, if uniformly distributed in a 10-m-thick layer, would be approximately 1 cell per ml. PMID- 16339016 TI - Beyond the olfactory bulb: an odotopic map in the forebrain. AB - We report electrophysiological evidence that a simple odotopy, the spatial mapping of different odorants, is maintained above the level of the olfactory bulb (OB). Three classes of biologically relevant odorants for fish are processed in distinct regions of the forebrain (FB) in the channel catfish. Feeding cues, mainly amino acids and nucleotides, are represented in lateral, pallial portions of the FB, equivalent to the olfactory cortex of amniote vertebrates, whereas social signals mediated by bile salts are represented in medial FB centers, possibly homologous to portions of the amygdala. As in the OB, the different odorant classes map onto different territories; however, the response properties of units of the olfactory areas of the FB do not simply mirror those of the OB. For some units, distinctive response properties emerged, because the FB is the first center where odors subserving a common behavioral function (i.e., food function) converge. PMID- 16339017 TI - Tandem subunits effectively constrain GABAA receptor stoichiometry and recapitulate receptor kinetics but are insensitive to GABAA receptor-associated protein. AB - GABAergic synapses likely contain multiple GABAA receptor subtypes, making postsynaptic currents difficult to dissect. However, even in heterologous expression systems, analysis of receptors composed of alpha, beta, and gamma subunits can be confounded by receptors expressed from alpha and beta subunits alone. To produce recombinant GABAA receptors containing fixed subunit stoichiometry, we coexpressed individual subunits with a "tandem" alpha1 subunit linked to a beta2 subunit. Cotransfection of the gamma2 subunit with alphabeta tandem subunits in human embryonic kidney 293 cells produced currents that were similar in their macroscopic kinetics, single-channel amplitudes, and pharmacology to overexpression of the gamma subunit with nonlinked alpha1 and beta2 subunits. Similarly, expression of alpha subunits together with alphabeta tandem subunits produced receptors having physiological and pharmacological characteristics that closely matched cotransfection of alpha with beta subunits. In this first description of tandem GABAA subunits measured with patch-clamp and rapid agonist application techniques, we conclude that incorporation of alphabeta tandem subunits can be used to fix stoichiometry and to establish the intrinsic kinetic properties of alpha1beta2 and alpha1beta2gamma2 receptors. We used this method to test whether the accessory protein GABAA receptor-associated protein (GABARAP) alters GABAA receptor properties directly or influences subunit composition. In recombinant receptors with fixed stoichiometry, coexpression of GABARAP-enhanced green fluorescent protein (EGFP) fusion protein had no effect on desensitization, deactivation, or diazepam potentiation of GABA-mediated currents. However, in alpha1beta2gamma2S transfections in which stoichiometry was not fixed, GABARAP-EGFP altered desensitization, deactivation, and diazepam potentiation of GABA-mediated currents. The data suggest that GABARAP does not alter receptor kinetics directly but by facilitating surface expression of alphabetagamma receptors. PMID- 16339018 TI - Contribution of 5-HT2 receptor subtypes to sleep-wakefulness and respiratory control, and functional adaptations in knock-out mice lacking 5-HT2A receptors. AB - Serotonin (5-hydroxytryptamine; 5-HT) plays key roles in sleep-wakefulness regulation. Evidence indicates that 5-HT2 receptors are involved mainly in non rapid eye movement sleep (NREMS) regulation and respiratory control. Here, we investigated the relative contribution of 5-HT(2A), 5-HT(2B), and 5-HT(2C) receptor subtypes to NREMS and breathing during sleep, using 5-HT2 subtype selective ligands in wild-type (5-HT(2A)+/+) and knock-out (5-HT(2A)-/-) mice that do not express 5-HT(2A) receptors. Acute blockade of 5-HT(2A) receptors induced an increase in NREMS in 5-HT(2A)+/+ mice, but not 5-HT(2A)-/- mutants, which spontaneously expressed less NREMS than wild-type animals. In 5-HT(2A)+/+ mice, 5-HT(2B) receptor blockade produced a reduction of NREMS, whereas receptor activation induced an increase in this sleep stage. These effects were less pronounced in 5-HT(2A)-/- mice, indicating a lower sensitivity of 5-HT(2B) receptors in mutants, with no change in 5-HT(2B) mRNA. Blockade of 5-HT(2C) receptors had no effect on NREMS in both strains. In addition, an increase in EEG power density after sleep deprivation was observed in 5-HT(2A)+/+ mice but not in 5-HT(2A)-/- mice. Whole-body plethysmographic recordings indicated that 5-HT(2A) receptor blockade in 5-HT(2A)+/+ mice reduced NREMS apneas and bradypneas that occurred after sighs. In contrast, in 5-HT(2A)-/- mutants, NREMS apneas were not modified, and bradypnea after sighs were more pronounced. Our results demonstrate that 5-HT exerts a 5-HT(2B)-mediated facilitation of NREMS, and an influence respectively inhibitory on NREMS and facilitatory on sleep apnea generation, via 5-HT(2A) receptors. Moreover, 5-HT(2A) gene knock-out leads to functional compensations yielding adaptive changes opposite to those caused by pharmacological blockade of 5-HT(2A) receptors in 5-HT(2A)+/+ mice. PMID- 16339019 TI - Recognition of familiar individuals in golden hamsters: a new method and functional neuroanatomy. AB - The ability to recognize individuals is essential for many aspects of social interaction and social organization, yet we know relatively little about the neural mechanisms underlying this ability. Most laboratory studies of individual recognition in rodents have studied differential responses to familiar versus unfamiliar individuals rather than differential responses to equally well known individuals having different significance for the subject. In experiment 1, we use a new method for studying true individual recognition in which male hamsters first had different experiences with two stimulus males (exposures to one male across a wire-mesh barrier and fights with another male). One day later, losers of fights were tested in a Y-maze for reactions to one of the two familiar males. Subjects tested with the familiar winner avoided this stimulus male, but subjects tested with the familiar, neutral male were attracted to him. Immunohistochemistry for c-Fos and Egr-1 implicate several areas of the brain in individual recognition, particularly the anterior piriform cortex, the CA1 and CA3 regions of anterior dorsal hippocampus, anterior and posterior dentate gyrus, and perirhinal cortex. In experiment 2, temporary inactivation of the CA1 region of anterior dorsal hippocampus by microinfusion of lidocaine eliminated the avoidance of the familiar winner, but a saline control injection had no effect. These results are the first to use a rodent model to characterize neural circuits involved in the recognition of equally well known individuals and the corresponding emotional responses to them. PMID- 16339020 TI - Motor memory consolidation in sleep shapes more effective neuronal representations. AB - Learning a motor skill involves a latent process of consolidation that develops after training to enhance the skill in the absence of any practice and crucially depends on sleep. Here, we show that this latent consolidation during sleep changes the brain representation of the motor skill by reducing overall the neocortical contributions to the representation. Functional magnetic resonance brain imaging was performed during initial training and 48 h later, at retesting, on a sequential finger movement task with training followed by either a night of regular sleep or sleep deprivation. An additional night of sleep for all subjects served to rule out unspecific effects of sleep loss at retrieval testing. Posttraining sleep, but not sleep deprivation, led to improved motor skill performance at retrieval. This sleep-dependent improvement was linked to greatly reduced brain activation in prefrontal, premotor, and primary motor cortical areas, along with a stronger involvement of left parietal cortical regions. Our findings indicate that storing a motor skill during sleep reorganizes its brain representation toward enhanced efficacy. PMID- 16339021 TI - Decreased neuronal death in Na+/H+ exchanger isoform 1-null mice after in vitro and in vivo ischemia. AB - Na+/H+ exchanger isoform 1 (NHE1) is a major acid extrusion mechanism after intracellular acidosis. We hypothesized that stimulation of NHE1 after cerebral ischemia contributes to the disruption of Na+ homeostasis and neuronal death. In the present study, expression of NHE1 was detected in cultured mouse cortical neurons. Three hours of oxygen and glucose deprivation (OGD) followed by 21 h of reoxygenation (REOX) led to 68 +/- 10% cell death. Inhibition of NHE1 with the potent inhibitor cariporide (HOE 642) or genetic ablation of NHE1 reduced OGD induced cell death by approximately 40-50% (p < 0.05). In NHE1(+/+) neurons, OGD caused a twofold increase in [Na+]i, and 60 min REOX triggered a sevenfold increase. Genetic ablation of NHE1 or HOE 642 treatment had no effects on the OGD mediated initial Na+(i) rise but reduced the second phase of Na+(i) rise by approximately 40-50%. In addition, 60 min REOX evoked a 1.5-fold increase in [Ca2+]i in NHE1(+/+) neurons, which was abolished by inhibition of either NHE1 or reverse-mode operation of Na+/Ca2+ exchange. OGD/REOX-mediated mitochondrial Ca2+ accumulation and cytochrome c release were attenuated by inhibition of NHE1 activity. In an in vivo focal ischemic model, 2 h of left middle cerebral artery occlusion followed by 24 h of reperfusion induced 84.8 +/- 8.0 mm3 infarction in NHE1(+/+) mice. NHE1(+/+) mice treated with HOE 642 or NHE1 heterozygous mice exhibited a approximately 33% decrease in infarct size (p < 0.05). These results imply that NHE1 activity disrupts Na+ and Ca2+ homeostasis and contributes to ischemic neuronal damage. PMID- 16339022 TI - Synaptic dynamics do not determine proper phase of activity in a central pattern generator. AB - Rhythmic motor activity often requires neuronal output to the muscles to arrive in a particular sequence. At the pattern-generator level, this requires distinct activity phases in different groups of constituent neurons. The phase differences between rhythmically active neurons in a network are thought to arise from the interplay between their intrinsic properties and the temporal dynamics of synapses among these neurons. In the rhythmically active pyloric network of the lobster Panulirus interruptus, synaptic connections from the pacemaker ensemble to the follower neurons [lateral pyloric (LP) and pyloric constrictor (PY)] are thought to be primarily responsible for the proper phase of activity (pacemaker LP-PY) across all frequencies (0.5-2 Hz) of the pyloric rhythm. We test this hypothesis by characterizing the synapses from the pacemaker ensemble to the LP and PY neurons. Paired comparisons show that these two synapses are not significantly different in strength or in the extent of short-term depression. To examine the level to which intrinsic properties of the follower neurons determine their relative activity phase, we block all chemical synapses within the network and drive the LP and PY neurons rhythmically using artificial synaptic currents with identical strength and dynamics implemented with the dynamic-clamp technique. In response to these identical synaptic inputs, the LP and PY neurons maintain the proper relative phase of activity. These results strongly indicate that the relative phase of activity among these follower neurons within the pyloric network is not dictated by their synaptic inputs but is solely determined by their distinct intrinsic properties. PMID- 16339023 TI - Bidirectional activity-dependent plasticity at corticostriatal synapses. AB - Corticostriatal projections originate from the entire cerebral cortex and provide the major source of glutamatergic inputs to the basal ganglia. Despite the importance of corticostriatal connections in sensorimotor learning and cognitive functions, plasticity forms at these synapses remain strongly debated. Using a corticostriatal slice preserving the connections between the somatosensory cortex and the target striatal cells, we report the induction of both non-Hebbian and Hebbian forms of long-term potentiation (LTP) and long-term depression (LTD) on striatal output neurons (SONs). LTP and LTD can be induced selectively by different stimulation patterns (high-frequency trains vs low-frequency pulses) and were evoked with similar efficiency in non-Hebbian and Hebbian modes. Combination of LTP-LTD and LTD-LTP sequences revealed that bidirectional plasticity occurs at the same SONs and provides efficient homeostatic mechanisms leading to a resetting of corticostriatal synapses avoiding synaptic saturation. The effect of temporal relationship between cortical stimulation and SON activity was assessed using spike-timing-dependent plasticity (STDP) protocols. An LTP was observed when an action potential was triggered in the striatal neuron before the cortical stimulus, and, conversely, an LTD was induced when the striatal neuron discharge was triggered after the cortical stimulation. Such STDP was reversed when compared with those described so far in other mammalian brain structures. This mechanism may be essential for the role of the striatum in learning of motor sequences in which sensory and motor events are associated in a precise time sequence. PMID- 16339024 TI - Regulation of dendritic morphogenesis by Ras-PI3K-Akt-mTOR and Ras-MAPK signaling pathways. AB - Dendritic arborization and spine formation are critical for the functioning of neurons. Although many proteins have been identified recently as regulators of dendritic morphogenesis, the intracellular signaling pathways that control these processes are not well understood. Here we report that the Ras phosphatidylinositol 3-kinase (PI3K)-Akt-mammalian target of rapamycin (mTOR) signaling pathway plays pivotal roles in the regulation of many aspects of dendrite formation. Whereas the PI3K-Akt-mTOR pathway alone controlled soma and dendrite size, a coordinated activation together with the Ras-mitogen-activated protein kinase signaling pathway was required for increasing dendritic complexity. Chronic inhibition of PI3K or mTOR reduced soma and dendrite size and dendritic complexity, as well as density of dendritic filopodia and spines, whereas a short-term inhibition promoted the formation of mushroom-shaped spines on cells expressing constitutively active mutants of Ras, PI3K, or Akt, or treated with the upstream activator BDNF. Together, our data underscore the central role of a spatiotemporally regulated key cell survival and growth pathway on trophic regulation of the coordinated development of dendrite size and shape. PMID- 16339025 TI - Control of dendritic arborization by the phosphoinositide-3'-kinase-Akt-mammalian target of rapamycin pathway. AB - The molecular mechanisms that determine the size and complexity of the neuronal dendritic tree are unclear. Here, we show that the phosphoinositide-3' kinase (PI3K)-Akt-mammalian target of rapamycin (mTOR) signaling pathway promotes the growth and branching of dendrites in cultured hippocampal neurons. Constitutively active mutants of Ras, PI3K, and Akt, or RNA interference (RNAi) knockdown of lipid phosphatase PTEN (phosphatase and tensin homolog deleted on chromosome Ten), induced growth and elaboration of dendrites that was blocked by mTOR inhibitor rapamycin and/or by overexpression of eIF-4E binding protein 1 (4E BP1), which inhibits translation of 5' capped mRNAs. The effect of PI3K on dendrites was lost in more mature neurons (>14 d in vitro). Dendritic complexity was reduced by inhibition of PI3K and by RNAi knockdown of mTOR or p70 ribosomal S6 kinase (p70S6K, an effector of mTOR). A rapamycin-resistant mutant of mTOR "rescued" the morphogenetic effects of PI3K in the presence of rapamycin. By regulating global and/or local protein translation, and as a convergence point for multiple signaling pathways, mTOR could play a central role in the control of dendrite growth and branching during development and in response to activity. PMID- 16339026 TI - Beta-secretase-cleaved amyloid precursor protein accumulates at actin inclusions induced in neurons by stress or amyloid beta: a feedforward mechanism for Alzheimer's disease. AB - Rod-like inclusions (rods), composed of actin saturated with actin depolymerizing factor (ADF)/cofilin, are induced in hippocampal neurons by ATP depletion, oxidative stress, and excess glutamate and occur in close proximity to senile plaques in human Alzheimer's disease (AD) brain (Minamide et al., 2000). Here, we show rods are found in brains from transgenic AD mice. Soluble forms of amyloid beta (Abeta(1-42)) induce the formation of rods in a maximum of 19% of cultured hippocampal neurons in a time- and concentration-dependent manner. Approximately one-half of the responding neurons develop rods within 6 h or with as little as 10 nM Abeta(1-42). Abeta(1-42) induces the activation (dephosphorylation) of ADF/cofilin in neurons that form rods. Vesicles containing amyloid precursor protein (APP), beta-amyloid cleavage enzyme, and presenilin-1, a component of the gamma-secretase complex, accumulate at rods. The beta-secretase-cleaved APP (either beta-C-terminal fragment of APP or Abeta) also accumulates at rods. These results suggest that rods, formed in response to either Abeta or some other stress, block the transport of APP and enzymes involved in its processing to Abeta. These stalled vesicles may provide a site for producing Abeta(1-42), which may in turn induce more rods in surrounding neurons, and expand the degenerative zone resulting in plaque formation. PMID- 16339027 TI - Downregulation of transient receptor potential melastatin 8 by protein kinase C mediated dephosphorylation. AB - Transient receptor potential melastatin 8 (TRPM8) and transient receptor potential vanilloid 1 (TRPV1) are ion channels that detect cold and hot sensations, respectively. Their activation depolarizes the peripheral nerve terminals resulting in action potentials that propagate to brain via the spinal cord. These receptors also play a significant role in synaptic transmission between dorsal root ganglion (DRG) and dorsal horn (DH) neurons. Here, we show that TRPM8 is functionally downregulated by activation of protein kinase C (PKC) resulting in inhibition of membrane currents and increases in intracellular Ca2+ compared with upregulation of TRPV1 in cloned and native receptors. Bradykinin significantly downregulates TRPM8 via activation of PKC in DRG neurons. Activation of TRPM8 or TRPV1 at first sensory synapse between DRG and DH neurons leads to a robust increase in frequency of spontaneous/miniature EPSCs. PKC activation blunts TRPM8- and facilitates TRPV1-mediated synaptic transmission. Significantly, downregulation is attributable to PKC-mediated dephosphorylation of TRPM8 that could be reversed by phosphatase inhibitors. These findings suggest that inflammatory thermal hyperalgesia mediated by TRPV1 may be further aggravated by downregulation of TRPM8, because the latter could mediate the much needed cool/soothing sensation. PMID- 16339028 TI - Interaction of cellular prion and stress-inducible protein 1 promotes neuritogenesis and neuroprotection by distinct signaling pathways. AB - Understanding the physiological function of the cellular prion (PrPc) depends on the investigation of PrPc-interacting proteins. Stress-inducible protein 1 (STI1) is a specific PrPc ligand that promotes neuroprotection of retinal neurons through cAMP-dependent protein kinase A (PKA). Here, we examined the signaling pathways and functional consequences of the PrPc interaction with STI1 in hippocampal neurons. Both PrPc and STI1 are abundantly expressed and highly colocalized in the hippocampus in situ, indicating that they can interact in vivo. Recombinant STI1 (His6-STI1) added to hippocampal cultures interacts with PrPc at the neuronal surface and elicits neuritogenesis in wild-type neurons but not in PrPc-null cells. This effect was abolished by antibodies against either PrPc or STI1 and was dependent on the STI1 domain that binds PrPc. Binding of these proteins induced the phosphorylation/activation of the mitogen-activated protein kinase, which was essential for STI1-promoted neuritogenesis. His6-STI1, but not its counterpart lacking the PrPc binding site, prevented cell death via PKA activation. These results demonstrate that two parallel effects of the PrPc STI1 interaction, neuritogenesis and neuroprotection, are mediated by distinct signaling pathways. PMID- 16339029 TI - Neural mechanisms underlying probabilistic category learning in normal aging. AB - Probabilistic category learning engages neural circuitry that includes the prefrontal cortex and caudate nucleus, two regions that show prominent changes with normal aging. However, the specific contributions of these brain regions are uncertain, and the effects of normal aging have not been examined previously in probabilistic category learning. In the present study, using a blood oxygenation level-dependent functional magnetic resonance imaging block design, 18 healthy young adults (mean age, 25.5 +/- 2.6 years) and 15 older adults (mean age, 67.1 +/- 5.3 years) were assessed on the probabilistic category learning "weather prediction" test. Whole-brain functional images acquired using a 1.5T scanner (General Electric, Milwaukee, WI) with gradient echo, echo planar imaging (3/1 mm; repetition time, 3000 ms; echo time, 50 ms) were analyzed using second-level random-effects procedures [SPM99 (Statistical Parametric Mapping)]. Young and older adults displayed equivalent probabilistic category learning curves, used similar strategies, and activated analogous neural networks, including the prefrontal and parietal cortices and the caudate nucleus. However, the extent of caudate and prefrontal activation was less and parietal activation was greater in older participants. The percentage correct and reaction time were mainly positively correlated with caudate and prefrontal activation in young individuals but positively correlated with prefrontal and parietal cortices in older individuals. Differential activation within a circumscribed neural network in the context of equivalent learning suggests that some brain regions, such as the parietal cortices, may provide a compensatory mechanism for healthy older adults in the context of deficient prefrontal cortex and caudate nuclei responses. PMID- 16339030 TI - Activation of gonadotropin-releasing hormone neurons by kisspeptin as a neuroendocrine switch for the onset of puberty. AB - We examined the role of kisspeptin and its receptor, the G-protein-coupled receptor GPR54, in governing the onset of puberty in the mouse. In the adult male and female mouse, kisspeptin (10-100 nM) evoked a remarkably potent, long-lasting depolarization of >90% of gonadotropin-releasing hormone (GnRH)-green fluorescent protein neurons in situ. In contrast, in juvenile [postnatal day 8 (P8) to P19] and prepubertal (P26-P33) male mice, kisspeptin activated only 27 and 44% of GnRH neurons, respectively. This developmental recruitment of GnRH neurons into a kisspeptin-responsive pool was paralleled by an increase in the ability of centrally administered kisspeptin to evoke luteinizing hormone secretion in vivo. To learn more about the mechanisms through which kisspeptin-GPR54 signaling at the GnRH neuron may change over postnatal development, we performed quantitative in situ hybridization for kisspeptin and GPR54 transcripts. Approximately 90% of GnRH neurons were found to express GPR54 mRNA in both juvenile and adult mice, without a detectable difference in the mRNA content between the age groups. In contrast, the expression of KiSS-1 mRNA increased dramatically across the transition from juvenile to adult life in the anteroventral periventricular nucleus (AVPV; p < 0.001). These results demonstrate that kisspeptin exerts a potent depolarizing effect on the excitability of almost all adult GnRH neurons and that the responsiveness of GnRH neurons to kisspeptin increases over postnatal development. Together, these observations suggest that activation of GnRH neurons by kisspeptin at puberty reflects a dual process involving an increase in kisspeptin input from the AVPV and a post-transcriptional change in GPR54 signaling within the GnRH neuron. PMID- 16339031 TI - Competitive stimulus interactions within single response fields of superior colliculus neurons. AB - In addition to its role in saccade generation, the superior colliculus (SC) is involved in target selection, saccade selection, and shifting the focus of spatial attention. Here, we investigated the influence of saccade selection on sensory interactions within single response fields (RFs) of SC neurons. One or two differently shaped stimuli were presented within single RFs of SC neurons, and the shape of a centrally located cue indicated whether and where to make a saccade (Go-Go) or whether to make or withhold a saccade (Go/No-Go). We found that, when two stimuli appeared at different locations within a single RF, SC neuronal activity was reduced compared with when a single stimulus appeared in isolation within the center of the RF in both the Go-Go and Go/No-Go tasks. In both tasks, a subsequent cue indicating one stimulus as a saccade target reduced the influence of the second stimulus located within the RF. We found that the time course of the suppression resulting from the two stimuli was approximately 130 ms, a time close to that seen in cortex. Finally, we found that the influence of two stimuli within single RFs of SC neurons changed over time in both the Go Go and the Go/No-Go tasks. Initially, the neurons averaged the influence of two stimuli. As the trial progressed, the SC neurons signaled only the saccade vector that was produced. We conclude that cues to shift gaze, like attention, modulate the influence of sensory interactions, providing additional support for the linkage between attention and saccade selection. PMID- 16339032 TI - Modulation of NMDA receptors by pituitary adenylate cyclase activating peptide in CA1 neurons requires G alpha q, protein kinase C, and activation of Src. AB - At CA1 synapses, activation of NMDA receptors (NMDARs) is required for the induction of both long-term potentiation and depression. The basal level of activity of these receptors is controlled by converging cell signals from G protein-coupled receptors and receptor tyrosine kinases. Pituitary adenylate cyclase activating peptide (PACAP) is implicated in the regulation of synaptic plasticity because it enhances NMDAR responses by stimulating Galphas-coupled receptors and protein kinase A (Yaka et al., 2003). However, the major hippocampal PACAP1 receptor (PAC1R) also signals via Galphaq subunits and protein kinase C (PKC). In CA1 neurons, we showed that PACAP38 (1 nM) enhanced synaptic NMDA, and evoked NMDAR, currents in isolated CA1 neurons via activation of the PAC1R, Galphaq, and PKC. The signaling was blocked by intracellular applications of the Src inhibitory peptide Src(40-58). Immunoblots confirmed that PACAP38 biochemically activates Src. A Galphaq pathway is responsible for this Src dependent PACAP enhancement because it was attenuated in mice lacking expression of phospholipase C beta1, it was blocked by preventing elevations in intracellular Ca2+, and it was eliminated by inhibiting either PKC or cell adhesion kinase beta [CAKbeta or Pyk2 (proline rich tyrosine kinase 2)]. Peptides that mimic the binding sites for either Fyn or Src on receptor for activated C kinase-1 (RACK1) also enhanced NMDAR in CA1 neurons, but their effects were blocked by Src(40-58), implying that Src is the ultimate regulator of NMDARs. RACK1 serves as a hub for PKC, Fyn, and Src and facilitates the regulation of basal NMDAR activity in CA1 hippocampal neurons. PMID- 16339033 TI - Noradrenaline triggers multivesicular release at glutamatergic synapses in the hypothalamus. AB - The origin of large-amplitude miniature EPSCs (mEPSCs) at central synapses remains to be firmly established. Here, we show that at excitatory synapses onto magnocellular neurosecretory cells in the hypothalamus, noradrenaline induces a rapid and robust increase in mEPSC amplitude that requires alpha1-adrenoceptor activation but is impervious to postsynaptic manipulations that block the putative insertion of AMPA receptors. In response to noradrenaline, mEPSCs exhibit a putative multimodal amplitude histogram distribution that is not attributable to random temporal summation, the unveiling of a quiescent synapse, or the release of large vesicles. Large-amplitude mEPSCs are sensitive to a high dose of ryanodine and are associated with an enhanced glutamate cleft concentration. Together, these data are consistent with the hypothesis that large amplitude mEPSCs result from the synchronous release of multiple vesicles via rapid presynaptic calcium expulsion from intracellular stores. PMID- 16339034 TI - Novel seizure phenotype and sleep disruptions in knock-in mice with hypersensitive alpha 4* nicotinic receptors. AB - A leucine to alanine substitution (L9'A) was introduced in the M2 region of the mouse alpha4 neuronal nicotinic acetylcholine receptor (nAChR) subunit. Expressed in Xenopus oocytes, alpha4(L9'A)beta2 nAChRs were > or =30-fold more sensitive than wild type (WT) to both ACh and nicotine. We generated knock-in mice with the L9'A mutation and studied their cellular responses, seizure phenotype, and sleep wake cycle. Seizure studies on alpha4-mutated animals are relevant to epilepsy research because all known mutations linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) occur in the M2 region of alpha4or beta2 subunits. Thalamic cultures and synaptosomes from L9'A mice were hypersensitive to nicotine induced ion flux. L9'A mice were approximately 15-fold more sensitive to seizures elicited by nicotine injection than their WT littermates. Seizures in L9'A mice differed qualitatively from those in WT: L9'A seizures started earlier, were prevented by nicotine pretreatment, lacked EEG spike-wave discharges, and consisted of fast repetitive movements. Nicotine-induced seizures in L9'A mice were partial, whereas WT seizures were generalized. When L9'A homozygous mice received a 10 mg/kg nicotine injection, there was temporal and phenomenological separation of mutant and WT-like seizures: an initial seizure approximately 20 s after injection was clonic and showed no EEG changes. A second seizure began 3-4 min after injection, was tonic-clonic, and had EEG spike-wave activity. No spontaneous seizures were detected in L9'A mice during chronic video/EEG recordings, but their sleep-wake cycle was altered. Our findings show that hypersensitive alpha4* nicotinic receptors in mice mediate changes in the sleep wake cycle and nicotine-induced seizures resembling ADNFLE. PMID- 16339035 TI - Prolonged reciprocal signaling via NMDA and GABA receptors at a retinal ribbon synapse. AB - AMPA and GABAA receptors mediate most of the fast signaling in the CNS. However, the retina must, in addition, also convey slow and sustained signals. Given that AMPA and GABAA receptors desensitize quickly in the continuous presence of agonist, how are sustained excitatory and inhibitory signals transmitted reliably across retinal synapses? Reciprocal synapses between bipolar and amacrine cells in the retina are thought to play a fundamental role in tuning the bipolar cell output to the dynamic range of ganglion cells. Here, we report that glutamate release from goldfish bipolar cell terminals activates first AMPA receptors, followed by fast and transient GABAA-mediated feedback. Subsequently, prolonged NMDA receptor activation triggers GABAA and a slow, sustained GABAC-mediated reciprocal inhibition. The synaptic delay of the NMDA/GABAC-mediated feedback showed stronger dependence on the depolarization of the bipolar cell terminal than the fast AMPA/GABAA-mediated response. Although the initial depolarization mediated by AMPA receptors was important to prime the NMDA action, NMDA receptors could trigger feedback by themselves in most of the bipolar terminals tested. This AMPA-independent feedback (delay approximately 10 ms) was eliminated in 2 mm external Mg2+ and reduced in some terminals, but not eliminated, by TTX. NMDA receptors on amacrine cells with depolarized resting membrane potentials therefore can mediate the late reciprocal feedback triggered by continuous glutamate release. Our findings suggest that the characteristics of NMDA receptors (high agonist affinity, slow desensitization, and activation/deactivation kinetics) are well suited to match the properties of GABAC receptors, which thus provide part of the prolonged inhibition to bipolar cell terminals. PMID- 16339036 TI - Neuronal sensitivity to microsecond time disparities in the electrosensory system of Gymnarchus niloticus. AB - To perform the jamming avoidance response (JAR), the weakly electric fish Gymnarchus detects time disparities on the order of microseconds between electrosensory signals received by electroreceptors in different parts of the body surface. This paper describes time-disparity thresholds of output neurons of the electrosensory lateral line lobe (ELL), where the representation of timing information is converted from a time code to a firing-rate code. We recorded extracellular single-unit responses from pyramidal cells in the ELL to sinusoidally modulated time disparity with various depths (0-200 micros). Threshold sensitivity to time disparities measured in 123 units ranged from 0.5 to 100 micros and was < or =5 micros in 60% of the units. The units from pyramidal cells in the inner and outer cell layers of the ELL responded equally well to small time disparities. The neuronal thresholds to time disparities found in the ELL are comparable with those demonstrated in behavioral performance of the JAR. The sensitivity of ELL units to small time disparities was unaffected when the center of the cyclic time-disparity modulation was shifted over a wide range (up to 250 micros), indicating an adaptation mechanism for steady-state time disparities that preserves the sensitivity to small dynamic changes in time disparities. Phase-locked input neurons, which provide time information to the ELL by phase-locked firing of action potentials, did not adapt to steady-state time shifts of sensory signals. This suggests that the adaptation emerges within the ELL. PMID- 16339037 TI - Multiple receptors coupled to phospholipase C gate long-term depression in visual cortex. AB - Long-term depression (LTD) in sensory cortices depends on the activation of NMDA receptors. Here, we report that in visual cortical slices, the induction of LTD (but not long-term potentiation) also requires the activation of receptors coupled to the phospholipase C (PLC) pathway. Using immunolesions in combination with agonists and antagonists, we selectively manipulated the activation of alpha1 adrenergic, M1 muscarinic, and mGluR5 glutamatergic receptors. Inactivation of these PLC-coupled receptors prevents the induction of LTD, but only when the three receptors were inactivated together. LTD is fully restored by activating any one of them or by supplying intracellular D-myo-inositol-1,4,5 triphosphate (IP3). LTD was also impaired by intracellular application of PLC or IP3 receptor blockers, and it was absent in mice lacking PLCbeta1, the predominant PLC isoform in the forebrain. We propose that visual cortical LTD requires a minimum of PLC activity that can be supplied independently by at least three neurotransmitter systems. This essential requirement places PLC-linked receptors in a unique position to control the induction of LTD and provides a mechanism for gating visual cortical plasticity via extra-retinal inputs in the intact organism. PMID- 16339038 TI - Parsing molecular and behavioral effects of cocaine in mitogen- and stress activated protein kinase-1-deficient mice. AB - Although the induction of persistent behavioral alterations by drugs of abuse requires the regulation of gene transcription, the precise intracellular signaling pathways that are involved remain mainly unknown. Extracellular signal regulated kinase (ERK) is critical for the expression of immediate-early genes in the striatum in response to cocaine and Delta9-tetrahydrocannabinol and for the rewarding properties of these drugs. Here we show that in mice a single injection of cocaine (10 mg/kg) activates mitogen- and stress-activated protein kinase 1 (MSK1) in dorsal striatum and nucleus accumbens. Cocaine-induced phosphorylation of MSK1 threonine 581 and cAMP response element-binding protein (CREB) serine 133 (Ser133) were blocked by SL327, a drug that prevents ERK activation. Cocaine increased the acetylation of histone H4 lysine 5 and phosphorylation of histone H3 Ser10, demonstrating the existence of drug-induced chromatin remodeling in vivo. In MSK1 knock-out (KO) mice CREB and H3 phosphorylation in response to cocaine (10 mg/kg) were blocked, and induction of c-Fos and dynorphin was prevented, whereas the induction of Egr-1 (early growth response-1)/zif268/Krox24 was unaltered. MSK1-KO mice had no obvious neurological defect but displayed a contrasted behavioral phenotype in response to cocaine. Acute effects of cocaine and dopamine D1 or D2 agonists were unaltered. Sensitivity to low doses, but not high doses, of cocaine was increased in the conditioned place preference paradigm, whereas locomotor sensitization to repeated injections of cocaine was decreased markedly. Our results show that MSK1 is a major striatal kinase, downstream from ERK, responsible for the phosphorylation of CREB and H3 and is required specifically for the induction of c-Fos and dynorphin as well as for locomotor sensitization. PMID- 16339039 TI - Modifying the subunit composition of TASK channels alters the modulation of a leak conductance in cerebellar granule neurons. AB - Two-pore domain potassium (K2P) channel expression is believed to underlie the developmental emergence of a potassium leak conductance [IK(SO)] in cerebellar granule neurons (CGNs), suggesting that K2P function is an important determinant of the input conductance and resting membrane potential. To investigate the role that different K2P channels may play in the regulation of CGN excitability, we generated a mouse lacking TASK-1, a K2P channel known to have high expression levels in CGNs. In situ hybridization and real-time PCR studies in wild-type and TASK-1 knock-outs (KOs) demonstrated that the expression of other K2P channels was unaltered in CGNs. TASK-1 knock-out mice were healthy and bred normally but exhibited compromised motor performance consistent with altered cerebellar function. Whole-cell recordings from adult cerebellar slice preparations revealed that the resting excitability of mature CGNs was no different in TASK-1 KO and littermate controls. However, the modulation of IK(SO) by extracellular Zn2+, ruthenium red, and H+ was altered. The IK(SO) recorded from TASK-1 knock-out CGNs was no longer sensitive to alkalization and was blocked by Zn2+ and ruthenium red. These results suggest that a TASK-1-containing channel population has been replaced by a homodimeric TASK-3 population in the TASK-1 knock-out. These data directly demonstrate that TASK-1 channels contribute to the properties of IK(SO) in adult CGNs. However, TASK channel subunit composition does not alter the resting excitability of CGNs but does influence sensitivity to endogenous modulators such as Zn2+ and H+. PMID- 16339040 TI - Molecular and cellular diversity of neuronal G-protein-gated potassium channels. AB - Neuronal G-protein-gated potassium (GIRK) channels mediate the inhibitory effects of many neurotransmitters. Although the overlapping distribution of GIRK subunits suggests that channel composition varies in the CNS, little direct evidence supports the existence of structural or functional diversity in the neuronal GIRK channel repertoire. Here we show that the GIRK channels linked to GABAB receptors differed in two neuron populations. In the substantia nigra, GIRK2 was the principal subunit, and it was found primarily in dendrites of neurons in the substantia nigra pars compacta (SNc). Baclofen evoked prominent barium-sensitive outward current in dopamine neurons of the SNc from wild-type mice, but this current was completely absent in neurons from GIRK2 knock-out mice. In the hippocampus, all three neuronal GIRK subunits were detected. The loss of GIRK1 or GIRK2 was correlated with equivalent, dramatic reductions in baclofen-evoked current in CA1 neurons. Virtually all of the barium-sensitive component of the baclofen-evoked current was eliminated with the ablation of both GIRK2 and GIRK3, indicating that channels containing GIRK3 contribute to the postsynaptic inhibitory effect of GABAB receptor activation. The impact of GIRK subunit ablation on baclofen-evoked current was consistent with observations that GIRK1, GIRK2, and GABAB receptors were enriched in lipid rafts isolated from mouse brain, whereas GIRK3 was found primarily in higher-density membrane fractions. Altogether, our data show that different GIRK channel subtypes can couple to GABAB receptors in vivo. Furthermore, subunit composition appears to specify interactions between GIRK channels and organizational elements involved in channel distribution and efficient receptor coupling. PMID- 16339041 TI - Central oxytocin, vasopressin, and corticotropin-releasing factor receptor densities in the basal forebrain predict isolation potentiated startle in rats. AB - Individual differences in resiliency to particular stressors may be mediated by specific neuropeptide receptor patterns in the brain. Here, we explored this issue by using a multivariate approach to identify brain sites in which oxytocin (OTR), vasopressin (V1aR), and corticotropin-releasing factor type 1 (CRF1) or type 2 receptor binding covaried with a measure of isolation-induced anxiety: isolation potentiated startle (IPS). Partial least squares (PLS) analysis identified three binding sites, the shell of the nucleus accumbens (AccSh), lateral bed nucleus of the stria terminalis, and intermediate zone of the lateral septum, in which CRF1, V1aR, and OTR receptors, respectively, covaried with IPS. Multiple regression analysis demonstrated that the three binding sites accounted for more of the variation in IPS as a linear combination than when considered individually. Using the same multiple regression model, the linear combination of the same three binding sites/peptide receptors measured in a new group of animals successfully predicted their IPS values. There were no differences in binding between grouped and isolated animals, suggesting that the patterns are trait effects rather than a consequence of isolation. Based on the finding that CRF1 receptors in the AccSh were positively correlated with IPS, we infused CRF directly into the AccSh and found that it significantly potentiated startle after a short isolation period but not under grouped conditions. This result directly supported the predictions made by the combined PLS/regression approach. These results suggest that the integrated activity of neuropeptide systems mediating both social behavior and anxiety underlie IPS. PMID- 16339042 TI - Oxytocin modulates neural circuitry for social cognition and fear in humans. AB - In non-human mammals, the neuropeptide oxytocin is a key mediator of complex emotional and social behaviors, including attachment, social recognition, and aggression. Oxytocin reduces anxiety and impacts on fear conditioning and extinction. Recently, oxytocin administration in humans was shown to increase trust, suggesting involvement of the amygdala, a central component of the neurocircuitry of fear and social cognition that has been linked to trust and highly expresses oxytocin receptors in many mammals. However, no human data on the effects of this peptide on brain function were available. Here, we show that human amygdala function is strongly modulated by oxytocin. We used functional magnetic resonance imaging to image amygdala activation by fear-inducing visual stimuli in 15 healthy males after double-blind crossover intranasal application of placebo or oxytocin. Compared with placebo, oxytocin potently reduced activation of the amygdala and reduced coupling of the amygdala to brainstem regions implicated in autonomic and behavioral manifestations of fear. Our results indicate a neural mechanism for the effects of oxytocin in social cognition in the human brain and provide a methodology and rationale for exploring therapeutic strategies in disorders in which abnormal amygdala function has been implicated, such as social phobia or autism. PMID- 16339043 TI - Contribution of germ cells to the differentiation and maturation of the ovary: insights from models of germ cell depletion. AB - In mammals, the role played by germ cells in ovarian differentiation and folliculogenesis has been the focus of an increasing number of studies over the last decades. From these studies, it has emerged that bidirectional communication between germ cells and surrounding companion cells is required as soon as the initial assembly of follicles. Models of germ cell depletion that arise from both spontaneous and experimentally induced mutations as well as irradiation or chemical treatments have been helpful in deciphering the role played by germ cells from the onset of ovarian differentiation onward. This review reports current knowledge and proposes novel hypotheses that can be formulated from these models about the contribution of germ cells to ovarian differentiation and folliculogenesis. In particular, it promotes the idea that the influence of germ cells on companion somatic cells varies within both ovarian differentiation and folliculogenesis. PMID- 16339044 TI - Channel-dependent permeation of water and glycerol in mouse morulae. AB - The cryosensitivity of mammalian embryos depends on the stage of development. Because permeability to water and cryoprotectants plays an important role in cryopreservation, it is plausible that the permeability is involved in the difference in the tolerance to cryopreservation among embryos at different developmental stages. In this study, we examined the permeability to water and glycerol of mouse oocytes and embryos, and tried to deduce the pathway for the movement of water and glycerol. The water permeability (L(P), microm min(-1) atm( 1)) of oocytes and four-cell embryos at 25 degrees C was low (0.63-0.70) and its Arrhenius activation energy (E(a), kcal/mol) was high (11.6-12.3), which implies that the water permeates through the plasma membrane by simple diffusion. On the other hand, the L(p) of morulae and blastocysts was quite high (3.6-4.5) and its E(a) was quite low (5.1-6.3), which implies that the water moves through water channels. Aquaporin inhibitors, phloretin and p-(chloromercuri) benzene sulfonate, reduced the L(p) of morulae significantly but not that of oocytes. By immunocytochemical analysis, aquaporin 3, which transports not only water but also glycerol, was detected in the morulae but not in the oocytes. Accordingly, the glycerol permeability (P(GLY), x 10(-3) cm/min) of oocytes was also low (0.01) and its E(a) was remarkably high (41.6), whereas P(GLY) of morulae was quite high (4.63) and its E(a) was low (10.0). Aquaporin inhibitors reduced the P(GLY) of morulae significantly. In conclusion, water and glycerol appear to move across the plasma membrane mainly by simple diffusion in oocytes but by facilitated diffusion through water channel(s) including aquaporin 3 in morulae. PMID- 16339045 TI - Bovine NALP5, NALP8, and NALP9 genes: assignment to a QTL region and the expression in adult tissues, oocytes, and preimplantation embryos. AB - A 3204-bp full-length cDNA of bovine NALP9 was cloned and its genomic organization was analyzed. The 2988-bp open reading frame covers 9 exons and encodes a deduced protein of 996 amino acids containing Pyrin, Nacht and leucine rich repeat domains like the human NALP gene family members. Mapping with the WGRH5000 panel and fluorescence in situ hybridization assigned NALP9 in close vicinity to BM2078 (LOD score 25.71; distance 0.0 cR5000) on bovine chromosome 18, BTA18q25-q26, within a previously identified QTL region for reproductive traits flanked by the bovine marker BM2078 and TGLA227. BAC contig analysis revealed that NALP9, NALP8, and NALP5 map in this QTL region. Temporospatial expression of these members of the NALP gene family was monitored. Among the adult tissues examined, transcripts of NALP8 and NALP9 were detected exclusively in testis and ovary, whereas transcripts of the NALP5 gene are limited to the ovary. The transcripts of NALP9, NALP8, and NALP5 were detected in oocytes before and after in vitro maturation and with a gradual decline from 2-cell to 8-cell stage, suggesting no reactivation at the time of bovine maternal to embryonic transition. Assignment to a QTL region for reproductive traits and preferential expression of NALP9, NALP8, and NALP5 in oocyte, germinal lineage, and gonad cells may suggest their functional relevance to reproduction and possible contribution to phenotypic variation. PMID- 16339046 TI - Uneven distribution of expressed sequence tag loci on maize pachytene chromosomes. AB - Examining the relationships among DNA sequence, meiotic recombination, and chromosome structure at a genome-wide scale has been difficult because only a few markers connect genetic linkage maps with physical maps. Here, we have positioned 1195 genetically mapped expressed sequence tag (EST) markers onto the 10 pachytene chromosomes of maize by using a newly developed resource, the RN-cM map. The RN-cM map charts the distribution of crossing over in the form of recombination nodules (RNs) along synaptonemal complexes (SCs, pachytene chromosomes) and allows genetic cM distances to be converted into physical micrometer distances on chromosomes. When this conversion is made, most of the EST markers used in the study are located distally on the chromosomes in euchromatin. ESTs are significantly clustered on chromosomes, even when only euchromatic chromosomal segments are considered. Gene density and recombination rate (as measured by EST and RN frequencies, respectively) are strongly correlated. However, crossover frequencies for telomeric intervals are much higher than was expected from their EST frequencies. For pachytene chromosomes, EST density is about fourfold higher in euchromatin compared with heterochromatin, while DNA density is 1.4 times higher in heterochromatin than in euchromatin. Based on DNA density values and the fraction of pachytene chromosome length that is euchromatic, we estimate that approximately 1500 Mbp of the maize genome is in euchromatin. This overview of the organization of the maize genome will be useful in examining genome and chromosome evolution in plants. PMID- 16339047 TI - Re: "Preterm birth: the interaction of traffic-related air pollution with economic hardship in Los Angeles neighborhoods". PMID- 16339048 TI - Re: "Lung cancer and indoor pollution from heating and cooking with solid fuels: the IARC international multicentre case-control study in Eastern/Central Europe and the United Kingdom". PMID- 16339049 TI - Reporting participation in epidemiologic studies: a survey of practice. AB - Self-selection bias may threaten the internal validity of epidemiologic studies. Studies with a low level of participation are particularly vulnerable to this bias, and commentators note apparent declines in participation in recent years. The authors therefore conducted a retrospective review to survey the practice of reporting participation in epidemiologic studies, to assess changes in participation over time, and to evaluate the impact of increased biologic specimen collection on participation. The authors abstracted selected study characteristics from 355 peer-reviewed, original, analytic-epidemiology research articles published from January 1 to April 30, 2003, in 10 high-impact general epidemiology, public health, and medical journals. At least some information regarding participation was provided in 59% of cross-sectional studies, 44% of case-control studies, and 32% of cohort studies. Participation appears to have declined during 1970-2003 for all study designs. Participation declined most steeply for controls in population-based, case-control studies (-1.86% per year, 95% confidence interval: -3.03, -0.69), with steeper declines after 1990. Proportionately more studies collected biologic specimens over time, particularly for cohort and case-control study designs (p(trend) = 0.06 and 0.03, respectively), yet participation was reported separately for the biologic specimen study component in only 27% of studies. The authors conclude that epidemiologists need to address declining participation and to report participation consistently, including for biologic specimen collection. PMID- 16339050 TI - Refined estimate of the incubation period of severe acute respiratory syndrome and related influencing factors. AB - Many epidemiologists have agreed that a refined estimate of the incubation period of severe acute respiratory syndrome (SARS) would need a sample size of about 200 cases and appropriate statistical methods enabling the inclusion of cases with defined periods of exposure. However, no such studies have been reported so far. Besides, determinants of the SARS incubation period remain unclear. In this study, 209 probable SARS cases with documented episodes of exposure between March 1 and May 31, 2003, in mainland China were included. A nonparametric method was used to analyze these data with defined periods of exposure to obtain the refined estimate of the SARS incubation period. Furthermore, the authors also explored the influence of various factors on the SARS incubation period by analysis of variance, linear regression analysis, and analysis of covariance. The estimates of mean and variance of the SARS incubation period were 5.29 days and 12.33 days(2), respectively; 90% of patients would have an incubation period of less than 11.58 days with a probability of 0.8, and 99% of patients would have an incubation of less than 22.22 days with a probability of 0.9. The affected area showed a highly significant effect on the incubation period (p < 0.001), but the contact pattern, occupation, gender, and age did not. PMID- 16339051 TI - The National Cancer Institute diet history questionnaire: validation of pyramid food servings. AB - The performance of the National Cancer Institute's food frequency questionnaire, the Diet History Questionnaire (DHQ), in estimating servings of 30 US Department of Agriculture Food Guide Pyramid food groups was evaluated in the Eating at America's Table Study (1997-1998), a nationally representative sample of men and women aged 20-79 years. Participants who completed four nonconsecutive, telephone administered 24-hour dietary recalls (n = 1,301) were mailed a DHQ; 965 respondents completed both the 24-hour dietary recalls and the DHQ. The US Department of Agriculture's Pyramid Servings Database was used to estimate intakes of pyramid servings for both diet assessment tools. The correlation (rho) between DHQ-reported intake and true intake and the attenuation factor (lambda) were estimated using a measurement error model with repeat 24-hour dietary recalls as the reference instrument. Correlations for energy-adjusted pyramid servings of foods ranged from 0.43 (other starchy vegetables) to 0.84 (milk) among women and from 0.42 (eggs) to 0.80 (total dairy food) among men. The mean rho and lambda after energy adjustment were 0.62 and 0.60 for women and 0.63 and 0.66 for men, respectively. This food frequency questionnaire validation study of foods measured in pyramid servings allowed for a measure of food intake consistent with national dietary guidance. PMID- 16339052 TI - A method for using incomplete triads to test maternally mediated genetic effects and parent-of-origin effects in relation to a quantitative trait. AB - The authors recently developed a semiparametric family-based test for linkage and association between markers and quantitative traits. This quantitative polytomous logistic regression test allows for analysis of families with incomplete information on parental genotype. In addition, it is not necessary to assume normality of the quantitative trait. Previous simulations have shown that the new test is as powerful as the other widely used tests for linkage disequilibrium in relation to a quantitative trait. Here the authors propose an extension to quantitative polytomous logistic regression that allows testing for maternally mediated effects and parent-of-origin effects in the same framework. Missing data on parental genotype are accommodated through an expectation-maximization algorithm approach. Simulations show robustness of the new tests and good power for detecting effects in the presence or absence of offspring effects. Methods are illustrated with birth weight and gestational length, two quantitative outcomes for which data were collected in a Montreal, Canada, study of intrauterine growth restriction between May 1998 and June 2000. PMID- 16339053 TI - Person-to-person transmission of hepatitis A virus in an urban area of intermediate endemicity: implications for vaccination strategies. AB - Developing countries with an increasing hepatitis A disease burden may target vaccination to specific groups, such as young children, as an initial control strategy. To better understand transmission of hepatitis A virus in such countries, the authors prospectively studied household and day-care/school contacts of cases in Almaty, Kazakhstan. Overall, by the time of identification of symptomatic index cases, half of transmission had already occurred, having been detected retrospectively. The odds of household contacts' becoming infected were 35.4 times those for day-care/school contacts (95% confidence interval (CI): 17.5, 71.7). Within households, younger age of either index cases or susceptible contacts elevated the odds of secondary infection among susceptible contacts: The presence of a case under 6 years of age raised the odds 4.7 times (95% CI: 1.2, 18.7); and compared with contacts aged 14 years or older, the odds of infection were increased to 7.7 (95% CI: 1.5, 40.3) and 7.0 (95% CI: 1.4, 34.3) among contacts aged 0-6 years and 7-13 years, respectively. Young children are appropriate targets for sustainable hepatitis A vaccination programs in areas undergoing hepatitis A epidemiologic transition. If vaccine is determined to be highly effective postexposure and if it is feasible, vaccinating household contacts could be a useful additional control strategy. PMID- 16339054 TI - Association of overweight with breast cancer survival. AB - The authors investigated the association between overweight at the time of or soon after cancer diagnosis and survival in a cohort of 1,455 breast cancer patients aged 25-64 years. The patients were recruited into the Shanghai Breast Cancer Study (Shanghai, China), a population-based case-control study, between August 1996 and March 1998. The median follow-up time for this cohort was 5.1 years (1996-2002) after breast cancer diagnosis, and 240 deaths were identified. Being overweight at cancer diagnosis or soon afterward, as measured by body mass index (BMI; weight (kg)/height (m)(2)), was associated with poorer overall survival and disease-free survival. Five-year survival rates were 86.5%, 83.8%, and 80.1% for subjects whose BMIs were <23.0, 23.0-24.9, and >or=25.0, respectively (p = 0.02); the corresponding 5-year disease-free survival rates were 81.9%, 78.1%, and 76.6% (p = 0.05). The inverse association between BMI and survival persisted after adjustment for age at diagnosis and other known prognostic factors for breast cancer, including disease stage. The authors found neither waist:hip ratio nor waist circumference to be independently associated with overall survival or disease-free survival. These results suggest that excess weight may be an independent predictor of breast cancer survival among Chinese women. PMID- 16339055 TI - Folate, vitamin B6, multivitamin supplements, and colorectal cancer risk in women. AB - The authors evaluated associations between intakes of folate and vitamin B(6) and colorectal cancer risk among women enrolled in a randomized trial on aspirin and vitamin E in disease prevention. At baseline (1992-1995), 37,916 US women aged >or=45 years who were free of cancer and cardiovascular disease provided dietary information. During an average of 10.1 years of follow-up (through February 20, 2004), 220 colorectal adenocarcinoma cases were documented. Total folate and vitamin B(6) intakes were not significantly associated with the risk of colorectal cancer. However, dietary intakes of folate and vitamin B(6) were significantly inversely associated with colorectal cancer risk among women who were not taking supplements containing folate and vitamin B(6). Multivariable relative risks among women in the highest quintiles of intake versus the lowest were 1.16 (95% confidence interval (CI): 0.76, 1.79) for total folate, 1.14 (95% CI: 0.77, 1.69) for total vitamin B(6), 0.46 (95% CI: 0.26, 0.81) for dietary folate, and 0.69 (95% CI: 0.41, 1.15) for dietary vitamin B(6). The use of multivitamin supplements was not related to colorectal cancer risk. These findings suggest that higher dietary intakes of folate and vitamin B(6) may reduce the risk of colorectal cancer in women. An alternative explanation is that other factors related to dietary intakes of folate and vitamin B(6) account for the inverse associations. PMID- 16339056 TI - Computer-assisted pattern recognition of autoantibody results. AB - Immunoassay-based anti-nuclear antibody (ANA) screens are increasingly used in the initial evaluation of autoimmune disorders, but these tests offer no "pattern information" comparable to the information from indirect fluorescence assay-based screens. Thus, there is no indication of "next steps" when a positive result is obtained. To improve the utility of immunoassay-based ANA screening, we evaluated a new method that combines a multiplex immunoassay with a k nearest neighbor (kNN) algorithm for computer-assisted pattern recognition. We assembled a training set, consisting of 1,152 sera from patients with various rheumatic diseases and non-diseased patients. The clinical sensitivity and specificity of the multiplex method and algorithm were evaluated with a test set that consisted of 173 sera collected at a rheumatology clinic from patients diagnosed by using standard criteria, as well as 152 age- and sex-matched sera from presumably healthy individuals (sera collected at a blood bank). The test set was also evaluated with a HEp-2 cell-based enzyme-linked immunosorbent assay (ELISA). Both the ELISA and multiplex immunoassay results were positive for 94% of the systemic lupus erythematosus (SLE) patients. The kNN algorithm correctly proposed an SLE pattern for 84% of the antibody-positive SLE patients. For patients with no connective tissue disease, the multiplex method found fewer positive results than the ELISA screen, and no disease was proposed by the kNN algorithm for most of these patients. In conclusion, the automated algorithm could identify SLE patterns and may be useful in the identification of patients who would benefit from early referral to a specialist, as well as patients who do not require further evaluation. PMID- 16339057 TI - Expression of interleukin-8 receptors (CXCR1 and CXCR2) in premenopausal women with recurrent urinary tract infections. AB - The migration of neutrophils through infected tissues is mediated by the CXC chemokines and its receptors (CXCR1 and CXCR2). It has been proposed that a CXCR1 deficiency could confer susceptibility to acute pyelonephritis in children. The objective of the study is to assess the surface expression of CXCR1 and CXCR2 and the existence of polymorphisms in the CXCR1 gene in premenopausal women with recurrent urinary tract infections. The study included 20 premenopausal women with recurrent urinary infections, with normal urinary tracts, and without diseases potentially associated with relapsing urinary infections and 30 controls without previous urinary infections. The levels of CXCR1 and CXCR2 expression on neutrophils were measured and analyzed by flow cytometry by measuring the mean fluorescence intensity (MFI) channel. The promoter and coding regions of the CXCR1 gene were analyzed for the presence of polymorphisms by a sequence-based typing method. Patients with recurrent urinary tract infections exhibited median levels of CXCR1 expression, determined from MFI values, similar to those of the controls. The analysis of CXCR2 showed that patients with recurrent urinary infections had lower median levels of expression, determined from the MFI values, than the controls (P = 0.002, Mann-Whitney U test). No polymorphisms were detected at the promoter or at the exon 1 region of the CXCR1 gene either in the patients or in the controls. Polymorphisms were detected at the exon 2 of CXCR1, but their frequencies did not differ between patients and controls. We have found a low level of CXCR2 expression in patients with recurrent urinary tract infections. These results suggest that a low level of CXCR2 expression may increase the susceptibilities of premenopausal women to urinary tract infections. PMID- 16339058 TI - Boosting immune response to hepatitis B DNA vaccine by coadministration of Prothymosin alpha-expressing plasmid. AB - DNA vaccines induce protective humoral and cell-mediated immune responses in several animal models. However, compared to conventional vaccines, DNA vaccines usually induce poor antibody responses. In this study, we report that coadministration of a hepatitis B virus (HBV) DNA vaccine with prothymosin alpha as an adjuvant improves antibody responses to HBV S antigen. We also observed higher seroconversion rates and higher antibody titers. Prothymosin alpha appears to increase the number and affinity of hepatitis B surface antigen-specific, gamma interferon-secreting T cells and to enhance cellular immune response to the PreS2S DNA vaccine. Interestingly, administering the DNA separately from the prothymosin alpha plasmid abrogated the enhancement of DNA vaccine potency. The results suggest that prothymosin alpha may be a promising adjuvant for DNA vaccines. PMID- 16339059 TI - Mycoplasma alligatoris infection promotes CD95 (FasR) expression and apoptosis of primary cardiac fibroblasts. AB - Mycoplasma alligatoris causes acute lethal primary infection of susceptible hosts. A genome survey implicated sialidase and hyaluronidase, potential promoters of CD95-mediated eukaryotic cell death, as virulence factors of M. alligatoris. We used immunofluorescence imaging and flow cytometry to examine the effects of M. alligatoris infection in vitro on CD95 expression and apoptosis by alligator cardiac fibroblasts, a major cell type of a target organ of M. alligatoris infection in vivo. A uniform distribution of CD95 in primary cultured cardiac, skeletal muscle, and embryonic fibroblasts was demonstrated by using polyclonal antibodies against the N or C terminus of mouse or human CD95. Anti CD95 antibodies reacted on Western blots of fibroblast lysates with a band with the predicted apparent molecular weight of CD95, but soluble CD95 was not detected in plasma from control or M. alligatoris-infected alligators. The proportion of CD95-gated cardiac fibroblasts increased threefold (P<0.01) 48 h after inoculation with M. alligatoris. Infection induced morphological changes in cardiac fibroblasts, including translocation of CD95 characteristic of apoptosis and an eightfold increase (P<0.16) in 5-bromo-2'-deoxyuridine (BrdU) incorporation measured in a terminal deoxynucleotide transferase dUTP nick end labeling apoptosis assay. The proportion of BrdU-gated controls activated with agonistic immunoglobulin M against human CD95 also increased threefold (P<0.03 for muscle). Heat-inactivated M. alligatoris and sterile M. alligatoris conditioned culture supernatant had no effect. This is the first report of a CD95 homolog in the class Reptilia and establishes a new model that can be used to test the direct bacterial interaction with upstream components of the CD95 signal transduction pathway. PMID- 16339060 TI - Lactobacillus johnsonii La1 attenuates Helicobacter pylori-associated gastritis and reduces levels of proinflammatory chemokines in C57BL/6 mice. AB - In clinical settings, Lactobacillus johnsonii La1 administration has been reported to have a favorable effect on Helicobacter pylori-associated gastritis, although the mechanism remains unclear. We administered, continuously through the water supply, live La1 to H. pylori-infected C57BL/6 mice and followed colonization, the development of H. pylori-associated gastritis in the lamina propria, and the levels of proinflammatory chemokines macrophage inflammatory protein 2 (MIP-2) and keratinocyte-derived cytokine (KC) in the serum and gastric tissue over a period of 3 months. We documented a significant attenuation in both lymphocytic (P=0.038) and neutrophilic (P=0.003) inflammatory infiltration in the lamina propria as well as in the circulating levels of anti-H. pylori immunoglobulin G antibodies (P=0.003), although we did not observe a suppressive effect of La1 on H. pylori colonizing numbers. Other lactobacilli, such as L. amylovorus DCE 471 and L. acidophilus IBB 801, did not attenuate H. pylori associated gastritis to the same extent. MIP-2 serum levels were distinctly reduced during the early stages of H. pylori infection in the La1-treated animals, as were gastric mucosal levels of MIP-2 and KC. Finally, we also observed a significant reduction (P=0.046) in H. pylori-induced interleukin-8 secretion by human adenocarcinoma AGS cells in vitro in the presence of neutralized (pH 6.8) La1 spent culture supernatants, without concomitant loss of H. pylori viability. These observations suggest that during the early infection stages, administration of La1 can attenuate H. pylori-induced gastritis in vivo, possibly by reducing proinflammatory chemotactic signals responsible for the recruitment of lymphocytes and neutrophils in the lamina propria. PMID- 16339061 TI - Modulation of antibody-mediated immune response by probiotics in chickens. AB - Probiotic bacteria, including Lactobacillus acidophilus and Bifidobacterium bifidum, have been shown to enhance antibody responses in mammals. The objective of this study was to examine the effects of a probiotic product containing the above bacteria in addition to Streptococcus faecalis on the induction of the chicken antibody response to various antigens, both systemically and in the gut. The birds received probiotics via oral gavage and subsequently were immunized with sheep red blood cells (SRBC) and bovine serum albumin (BSA) to evaluate antibody responses in serum or with tetanus toxoid (TT) to measure the mucosal antibody response in gut contents. Control groups received phosphate-buffered saline. Overall, BSA and SRBC induced a detectable antibody response as early as week 1 postimmunization (p.i.), which lasted until week 3 p.i. Probiotic-treated birds had significantly (P 90%). When the IgG1 ELISA was used annually over a 4-year period in a deer herd with high levels of clinical JD, it eliminated clinical disease, increased production levels, and reduced JD-related mortality. PMID- 16339062 TI - Local and systemic immune and inflammatory responses to Helicobacter pylori strains. AB - Colonization with Helicobacter pylori eventuates in varied clinical outcomes, which relate to both bacterial and host factors. Here we examine the relationships between cagA status, serum and gastric juice antibody responses, and gastric inflammation in dyspeptic patients. Serum, gastric juice, and gastric biopsy specimens were obtained from 89 patients undergoing endoscopy. H. pylori colonization and cagA status were determined by histology, culture, and PCR methods, and acute inflammation and chronic inflammation in the gastric mucosa were scored by a single pathologist. Serum and gastric juice antibodies to H. pylori whole-cell and CagA antigens were determined by enzyme-linked immunosorbent assay. Relationships between variables were sequentially analyzed using univariate and multivariate statistical methods. Of the 89 subjects, 62 were colonized by H. pylori. By univariate analyses, levels of serum immunoglobulin G (IgG) and IgA and gastric juice IgA antibodies against whole cell and CagA antigens each were significantly higher in the H. pylori-positive group than in the H. pylori-negative group (P<0.001). H. pylori and CagA sero positivities were both significantly associated with enhanced inflammation in gastric antrum and body (P<0.02). The presence of gastric juice antibodies to H. pylori antigens was associated with more severe gastric inflammation. However, in multivariate analyses, only the presence of serum antibodies against CagA and, to a lesser extent, whole-cell antigens remained significantly associated with acute and chronic inflammation in antrum and body (P<0.05). Thus, serum antibody response to CagA correlates with severity of gastric inflammation. Furthermore, given the relationships demonstrated by multivariate analysis, determination of gastric juice antibodies may provide a better representation of serum, rather than secretory, immune response. PMID- 16339064 TI - Application of an improved method for the recombinant k 39 enzyme-linked immunosorbent assay to detect visceral leishmaniasis disease and infection in Bangladesh. AB - Several serology-based immunoassays are used to diagnose visceral leishmaniasis (VL), a chronic protozoan parasitic disease caused by the Leishmania donovani complex. These tests are primarily designed to diagnose the most severe clinical form of VL, known as kala-azar. However, leishmanial infection is frequently asymptomatic and may manifest only as a positive serologic response or positive leishmanin skin test. We modified a previously described enzyme-linked immunosorbent assay (ELISA) that detects patient antibodies reactive with the recombinant Leishmania protein K39 (rK39) to confirm suspected kala-azar and to detect asymptomatic infection in a community study in Bangladesh. With the inclusion of a standard curve on each ELISA plate, the rK39 ELISA was more repeatable (kappa coefficient of agreement=0.970) and more reliable compared to the original method (kappa=0.587, P<0.001). The cutoff point for a positive antibody response was chosen based on the 99th percentile of the ELISA distribution for the negative-control sera. However, we found that sera from all patients with active kala-azar yielded values more than twice the magnitude of this cutoff. Using receiver-operator characteristic curves, we determined a second cutoff value predictive of kala-azar. Using these criteria, the sensitivity and specificity of the modified ELISA for kala-azar were 97.0% and 98.9%, respectively, for sera from our study population. We hypothesize that individuals with antibody levels greater than the 99th percentile of the negative controls but less than the cutoff point for kala-azar have asymptomatic leishmanial infections. PMID- 16339065 TI - Evaluation of a new single-parameter volumetric flow cytometer (CyFlow(green)) for enumeration of absolute CD4+ T lymphocytes in human immunodeficiency virus type 1-infected Thai patients. AB - Use of the standard dual-platform flow cytometric method for determination of CD4(+) T-lymphocyte counts, which needs both a flow cytometer (FCM) and hematological analyzer, would inevitably lead to increased variability. The development of new single-platform (SP) FCMs that provide direct CD4(+) T lymphocyte counts for improved assay precision and accuracy have recently attracted attention. This study evaluated one of those systems, CyFlow(green) (Partec), a single-parameter SP volumetric FCM. The performance of CyFlow(green) was compared with those of two reference standard SP microbead-based technologies of the three-color TruCOUNT tube with the FACScan FCM and a two-color FACSCount system (Becton Dickinson Biosciences). Absolute CD4(+) and CD8(+) T-lymphocyte counts in 200 human immunodeficiency virus type 1-seropositive blood specimens were determined. Statistical analysis for correlation and agreement were performed. A high correlation of absolute CD4 counts was shown when those obtained with CyFlow(green) were compared with those obtained with the bead-based three-color TruCOUNT system (R(2)=0.96; mean bias, -69.1 cells/microl; 95% confidence interval [CI], -225.7 to+87.5 cells/microl) and the FACSCount system (R(2)=0.97; mean bias, -40.0 cells/microl; 95% CI, -165.1 to+85.1 cells/microl). The correlation of the CD4(+) T-lymphocyte counts obtained by the two bead-based systems was high (R(2)=0.98). Interestingly, CyFlow(green) yielded CD4(+) T lymphocyte counts that were 21.8 and 7.2 cells/microl lower than those obtained with the TruCOUNT and the FACSCount systems, respectively, when CD4(+) T lymphocyte counts were <250 CD4(+) T-lymphocyte counts/microl range or 17.3 and 5.8 cells/microl less, respectively, when CD4(+) T-lymphocyte counts were <200 cells/microl. The single-parameter CyFlow(green) volumetric technology performed well in comparison with the performance of the standard SP bead-based FCM system. However, a multicenter comparative study is needed before this FCM machine is implemented in resource-limited settings. PMID- 16339066 TI - Evaluation and diagnostic usefulness of domestic and imported enzyme-linked immunosorbent assays for detection of human immunodeficiency virus type 1 antibody in India. AB - Diagnosis of human immunodeficiency virus (HIV) infection is important for patient management and prevention of new infections. The number of test kits available for the detection of HIV antibodies is unprecedented. In order to identify appropriate test kits, we evaluated a variety of commercial kits manufactured abroad as well as in India. The plasma and serum specimens (n=264) were collected from individuals attending the Voluntary Counseling and Testing Centre at the YRG Centre for AIDS and Education. The specimens were used to evaluate six commercially available HIV test kits: Enzaids HIV 1+2, HIV-CheX, Murex HIV-1.2.0, Genscreen HIV 1/2 version 2, Vironostika HIV Uni-Form II Ag/Ab, and CombAids RS Advantage. High sensitivities and specificities (>or=99%) were observed for the Enzaids, Murex, Vironostika, and CombAids assays. HIV-CheX showed the highest number of false-positive and false-negative results. The Genscreen test also gave many false positives. The study indicated that the Enzaids, Murex, and Vironostika enzyme-linked immunosorbent assay kits and the CombAids RS Advantage rapid assay could be used to achieve acceptable results for the detection of HIV antibodies. A combination of two tests is recommended to optimize the efficiency of HIV antibody testing algorithms, especially when evaluation with an HIV Western blot confirmatory test is not possible. PMID- 16339068 TI - Gamma interferon is dispensable for neopterin production in vivo. AB - Previous studies have indicated that neopterin is synthesized in vitro by human monocyte-derived macrophages and dendritic cells upon stimulation with gamma interferon (IFN-gamma). Neopterin production under specific conditions in vitro has also been obtained upon stimulation with IFN-alpha and/or IFN-beta. However, it is unknown if any IFN-gamma-independent neopterin synthesis is possible in vivo. In the present study we investigated the serum neopterin concentrations in patients affected by the syndrome of Mendelian susceptibility to mycobacterial disease (MSMD). Indeed, this syndrome is characterized by deeply impaired or absent IFN-gamma production or function due to severe mutations in molecules involved in IFN-gamma/interleukin-12 (IL-12)/IL-23-dependent pathway. Serum neopterin levels were measured by an enzyme-linked immunosorbent assay in 27 patients with MSMD. We found that serum neopterin levels are elevated in the complete absence of IFN-gamma activity due either to a complete deficiency of its receptor or to deleterious mutations of IL-12 or its receptor. These data clearly indicate that, as reported from in vitro studies, other stimuli are able to induce neopterin synthesis in vivo. Consequently, neopterin cannot be used as means of diagnosis of MSMD due to IFN-gamma-, IL-12-, and IL-23-dependent pathway defects. PMID- 16339067 TI - Induction of antigen-specific Th1-type immune responses by gamma-irradiated recombinant Brucella abortus RB51. AB - Brucella abortus strain RB51 is an attenuated rough mutant used as the live vaccine against bovine brucellosis in the United States and other countries. We previously reported the development of strain RB51 as a bacterial vaccine vector for inducing Th1-type immune responses against heterologous proteins. Because safety concerns may preclude the use of strain RB51-based recombinant live vaccines, we explored the ability of a gamma-irradiated recombinant RB51 strain to induce heterologous antigen-specific immune responses in BALB/c mice. Exposure of strain RB51G/LacZ expressing Escherichia coli beta-galactosidase to a minimum of 300 kilorads of gamma radiation resulted in complete loss of replicative ability. These bacteria, however, remained metabolically active and continued to synthesize beta-galactosidase. A single intraperitoneal inoculation of mice with 10(9) CFU equivalents of gamma-irradiated, but not heat-killed, RB51G/LacZ induced a beta-galactosidase-specific Th1-type immune response. Though no obvious differences were detected in immune responses to B. abortus-specific antigens, mice vaccinated with gamma-irradiated, but not heat-killed, RB51G/LacZ developed significant protection against challenge with virulent B. abortus. In vitro experiments indicated that gamma-irradiated and heat-killed RB51G/LacZ induced maturation of dendritic cells; however, stimulation with gamma-irradiated bacteria resulted in more interleukin-12 secretion. These results suggest that recombinant RB51 strains exposed to an appropriate minimum dose of gamma radiation are unable to replicate but retain their ability to stimulate Th1 immune responses against the heterologous antigens and confer protection against B. abortus challenge in mice. PMID- 16339069 TI - Long-term follow-up of Hepatitis B Surface antibody levels in subjects receiving universal Hepatitis B vaccination in infancy in an area of hyperendemicity: correlation between radioimmunoassay and enzyme immunoassay. AB - The aims of the present study were to determine (i) the long-term immunogenicity and the decay rate of hepatitis B virus (HBV) surface antibody (anti-HBs) from universal hepatitis B vaccination at infancy for a healthy population in an area of hyperendemicity and (ii) whether the anti-HBs levels measured by enzyme immunoassay (EIA) were closely correlated with those assayed by radioimmunoassay (RIA) methods during long-term monitoring. A total of 1,337 apparently healthy children (696 boys and 641 girls) who were vaccinated against HBV at infancy and monitored for anti-HBs annually from 7 to 16 years of age entered the study. Serum samples were analyzed for anti-HBs by RIA at 7 to 15 years of age and were also analyzed by EIA at 13 to 16 years of age. Antibody titers were quantified in mIU/ml by EIA as well as by the ratio of the count in the sample to the count for a negative control (S/N) by RIA. In non-boosted children, the average decay of anti-HBs from 7 to 16 years of ages indicated that approximately 20% of the geometric mean titer decays per year. There was a good correlation between serum anti-HBs levels measured by the RIA and the EIA methods (r=0.91; P<0.0001). An equation for RIA to EIA level conversion was established: log EIA titer=-0.12+ (1.31 . log RIA S/N). The anti-HBs titers measured by EIA correlate well with the S/N assayed by RIA. The annual decay rate of the log anti-HBs level may help in planning booster immunizations for hypo-responders or individuals at risk in adolescence. PMID- 16339070 TI - Unidirectional suppression of Anaplasma phagocytophilum genotypes in infected lambs. AB - Five-month-old lambs were simultaneously infected with different doses of two 16S rRNA genetic variants of Anaplasma phagocytophilum and thereafter followed for clinical observation and blood sampling. The result of the study indicates a unidirectional suppression of genotypes in infected lambs, at least during a certain period of an A. phagocytophilum infection. PMID- 16339071 TI - Immunoglobulin E binding reactivity of a recombinant allergen homologous to alpha Tubulin from Tyrophagus putrescentiae. AB - Storage mites may cause allergic respiratory diseases in urban areas as well as pose an occupational hazard in rural areas. Characterization of storage mite allergens is important for the development of diagnostic and therapeutic agents against mite-associated allergic disorders. Here we report on the cloning and expression of alpha-tubulin from the storage mite (Tyrophagus putrescentiae). The deduced amino acid sequence of the alpha-tubulin from the storage mite showed as much as 97.3% identity to the alpha-tubulin sequences from other organisms. The highly conserved amino acid sequences of alpha-tubulins across different species of mites may indicate that cross-reactivity for this potential allergen exists. The frequency of immunoglobulin E reactivity of this recombinant protein is 29.3% in sera from storage mite-allergic subjects. PMID- 16339072 TI - Longitudinal analysis of Severe Acute Respiratory Syndrome (SARS) coronavirus specific antibody in SARS patients. AB - The serum antibodies to severe acute respiratory syndrome (SARS) coronavirus of 18 SARS patients were checked at 1 month and every 3 months after disease onset. All of them except one, who missed blood sampling at 1 month, tested positive for the immunoglobulin G (IgG) antibody at 1 month. Fifteen out of 17 tested positive for the IgM antibody at 1 month. The serum IgM antibody of most patients became undetectable within 6 months after the onset of SARS. The IgG antibody of all 17 patients, whose serum was checked 1 year after disease onset, remained positive. PMID- 16339073 TI - Rootletin interacts with C-Nap1 and may function as a physical linker between the pair of centrioles/basal bodies in cells. AB - Rootletin, a major structural component of the ciliary rootlet, is located at the basal bodies and centrosomes in ciliated and nonciliated cells, respectively. Here we investigated its potential role in the linkage of basal bodies/centrioles and the mechanism involved in such linkages. We show that rootletin interacts with C-Nap1, a protein restricted at the ends of centrioles and functioning in centrosome cohesion in interphase cells. Their interaction in vivo is supported by their colocalization at the basal bodies/centrioles and coordinated association with the centrioles during the cell cycle. Ultrastructural examinations demonstrate that rootletin fibers connect the basal bodies in ciliated cells and are present both at the ends of and in between the pair of centrioles in nonciliated cells. The latter finding stands in contrast with C Nap1, which is present only at the ends of the centrioles. Transient expression of C-Nap1 fragments dissociated rootletin fibers from the centrioles, resulting in centrosome separation in interphase. Overexpression of rootletin in cells caused multinucleation, micronucleation, and irregularity of nuclear shape and size, indicative of defects in chromosome separation. These data suggest that rootletin may function as a physical linker between the pair of basal bodies/centrioles by binding to C-Nap1. PMID- 16339074 TI - Cell cycle-regulated trafficking of human telomerase to telomeres. AB - Telomerase synthesizes telomeres at the ends of human chromosomes during S phase. The results presented here suggest that telomerase activity may be regulated by intranuclear trafficking of the key components of the enzyme in human cells. We examined the subcellular localization of endogenous human telomerase RNA (hTR) and telomerase reverse transcriptase (hTERT) in HeLa cervical carcinoma cells. Throughout most of the cell cycle, we found that the two essential components of telomerase accumulate at intranuclear sites separate from telomeres. However, during S phase, both hTR and hTERT are specifically recruited to subsets of telomeres. The localization of telomerase to telomeres is dynamic, peaking at mid S phase. We also found complex associations of both hTR and hTERT with nucleoli and Cajal bodies during S phase, implicating both structures in the biogenesis and trafficking of telomerase. Our results mark the first observation of human telomerase at telomeres and provide a mechanism for the cell cycle-dependent regulation of telomere synthesis in human cells. PMID- 16339075 TI - Brd4 is required for recovery from antimicrotubule drug-induced mitotic arrest: preservation of acetylated chromatin. AB - The mammalian bromodomain protein Brd4 interacts with mitotic chromosomes by binding to acetylated histone H3 and H4 and is thought to play a role in epigenetic memory. Mitotic cells are susceptible to antimicrotubule drugs. These drugs activate multiple response pathways and arrest cells at mitosis. We found that Brd4 was rapidly released from chromosomes upon treatment with antimicrotubule drugs, including the reversible agent nocodazole. Yet, when nocodazole was withdrawn, Brd4 was reloaded onto chromosomes, and cells proceeded to complete cell division. However, cells in which a Brd4 allele was disrupted (Brd4+/-), and expressing only half of the normal Brd4 levels, were defective in reloading Brd4 onto chromosomes. Consequently, Brd4+/- cells were impaired in their ability to recover from nocodazole-induced mitotic arrest: a large fraction of +/- cells failed to reach anaphase after drug withdrawal, and those that entered anaphase showed an increased frequency of abnormal chromosomal segregation. The reloading defect observed in Brd4+/- cells coincided with selective hypoacetylation of lysine residues on H3 and H4. The histone deacetylase inhibitor trichostatin A increased global histone acetylation and perturbed nocodazole-induced Brd4 unloading. Brd4 plays an integral part in a cellular response to drug-induced mitotic stress by preserving a properly acetylated chromatin status. PMID- 16339076 TI - Contractile ring-independent localization of DdINCENP, a protein important for spindle stability and cytokinesis. AB - Dictyostelium DdINCENP is a chromosomal passenger protein associated with centromeres, the spindle midzone, and poles during mitosis and the cleavage furrow during cytokinesis. Disruption of the single DdINCENP gene revealed important roles for this protein in mitosis and cytokinesis. DdINCENP null cells lack a robust spindle midzone and are hypersensitive to microtubule depolymerizing drugs, suggesting that their spindles may not be stable. Furthermore DdCP224, a protein homologous to the microtubule-stabilizing protein TOGp/XMAP215, was absent from the spindle midzone of DdINCENP null cells. Overexpression of DdCP224 rescued the weak spindle midzone defect of DdINCENP null cells. Although not required for the localization of the myosin II contractile ring and subsequent formation of a cleavage furrow, DdINCENP is important for the abscission of daughter cells at the end of cytokinesis. Finally, we show that the localization of DdINCENP at the cleavage furrow is modulated by myosin II but it occurs by a mechanism different from that controlling the formation of the contractile ring. PMID- 16339077 TI - MAGI-1 is required for Rap1 activation upon cell-cell contact and for enhancement of vascular endothelial cadherin-mediated cell adhesion. AB - Rap1 is a small GTPase that regulates adherens junction maturation. It remains elusive how Rap1 is activated upon cell-cell contact. We demonstrate for the first time that Rap1 is activated upon homophilic engagement of vascular endothelial cadherin (VE-cadherin) at the cell-cell contacts in living cells and that MAGI-1 is required for VE-cadherin-dependent Rap1 activation. We found that MAGI-1 localized to cell-cell contacts presumably by associating with beta catenin and that MAGI-1 bound to a guanine nucleotide exchange factor for Rap1, PDZ-GEF1. Depletion of MAGI-1 suppressed the cell-cell contact-induced Rap1 activation and the VE-cadherin-mediated cell-cell adhesion after Ca2+ switch. In addition, relocation of vinculin from cell-extracellular matrix contacts to cell cell contacts after the Ca2+ switch was inhibited in MAGI-1-depleted cells. Furthermore, inactivation of Rap1 by overexpression of Rap1GAPII impaired the VE cadherin-dependent cell adhesion. Collectively, MAGI-1 is important for VE cadherin-dependent Rap1 activation upon cell-cell contact. In addition, once activated, Rap1 upon cell-cell contacts positively regulate the adherens junction formation by relocating vinculin that supports VE-cadherin-based cell adhesion. PMID- 16339078 TI - Hsp27 enhances recovery of splicing as well as rephosphorylation of SRp38 after heat shock. AB - A heat stress causes a rapid inhibition of splicing. Exogenous expression of Hsp27 did not prevent that inhibition but enhanced the recovery of splicing afterward. Another small heat shock protein, alphaB-crystallin, had no effect. Hsp27, but not alphaB-crystallin, also hastened rephosphorylation of SRp38 dephosphorylated a potent inhibitor of splicing-after a heat shock, although it did not prevent dephosphorylation by a heat shock. The effect of Hsp27 on rephosphorylation of SRp38 required phosphorylatable Hsp27. A Hsp90 client protein was required for the effect of Hsp27 on recovery of spicing and on rephosphorylation of SRp38. Raising the Hsp70 level by either a pre-heat shock or by exogenous expression had no effect on either dephosphorylation of SRp38 during heat shock or rephosphorylation after heat shock. The phosphatase inhibitor calyculin A prevented dephosphorylation of SRp38 during a heat shock and caused complete rephosphorylation of SRp38 after a heat shock, indicating that cells recovering from a heat shock are not deficient in kinase activity. Together our data show that the activity of Hsp27 in restoring splicing is not due to a general thermoprotective effect of Hsp27, but that Hsp27 is an active participant in the (de)phosphorylation cascade controlling the activity of the splicing regulator SRp38. PMID- 16339079 TI - Conventional kinesin mediates microtubule-microtubule interactions in vivo. AB - Conventional kinesin is a ubiquitous organelle transporter that moves cargo toward the plus-ends of microtubules. In addition, several in vitro studies indicated a role of conventional kinesin in cross-bridging and sliding microtubules, but in vivo evidence for such a role is missing. In this study, we show that conventional kinesin mediates microtubule-microtubule interactions in the model fungus Ustilago maydis. Live cell imaging and ultrastructural analysis of various mutants in Kin1 revealed that this kinesin-1 motor is required for efficient microtubule bundling and participates in microtubule bending in vivo. High levels of Kin1 led to increased microtubule bending, whereas a rigor mutation in the motor head suppressed all microtubule motility and promoted strong microtubule bundling, indicating that kinesin can form cross-bridges between microtubules in living cells. This effect required a conserved region in the C terminus of Kin1, which was shown to bind microtubules in vitro. In addition, a fusion protein of yellow fluorescent protein and the Kin1tail localized to microtubule bundles, further supporting the idea that a conserved microtubule binding activity in the tail of conventional kinesins mediates microtubule-microtubule interactions in vivo. PMID- 16339080 TI - Role of the Hog1 stress-activated protein kinase in the global transcriptional response to stress in the fungal pathogen Candida albicans. AB - The resistance of Candida albicans to many stresses is dependent on the stress activated protein kinase (SAPK) Hog1. Hence we have explored the role of Hog1 in the regulation of transcriptional responses to stress. DNA microarrays were used to characterize the global transcriptional responses of HOG1 and hog1 cells to three stress conditions that activate the Hog1 SAPK: osmotic stress, oxidative stress, and heavy metal stress. This revealed both stress-specific transcriptional responses and a core transcriptional response to stress in C. albicans. The core transcriptional response was characterized by a subset of genes that responded in a stereotypical manner to all of the stresses analyzed. Inactivation of HOG1 significantly attenuated transcriptional responses to osmotic and heavy metal stresses, but not to oxidative stress, and this was reflected in the role of Hog1 in the regulation of C. albicans core stress genes. Instead, the Cap1 transcription factor plays a key role in the oxidative stress regulation of C. albicans core stress genes. Our data show that the SAPK network in C. albicans has diverged from corresponding networks in model yeasts and that the C. albicans SAPK pathway functions in parallel with other pathways to regulate the core transcriptional response to stress. PMID- 16339081 TI - Syntaxins 3 and 4 are concentrated in separate clusters on the plasma membrane before the establishment of cell polarity. AB - Syntaxins 3 and 4 localize to the apical and basolateral plasma membrane, respectively, of epithelial cells where they mediate vesicle fusion. Here, we report that before establishment of cell polarity, syntaxins 3 and 4 are confined to mutually exclusive, submicron-sized clusters. Syntaxin clusters are remarkably uniform in size, independent of expression levels, and are distinct from caveolae and clathrin-coated pits. SNAP-23 partially colocalizes with both syntaxin 3 and 4 clusters. Deletion of the apical targeting signal of syntaxin 3 does not prevent sorting into clusters away from syntaxin 4. Syntaxin 3 and 4 cluster formation depends on different mechanisms because the integrity of syntaxin 3 clusters depends on intact microtubules, whereas syntaxin 4 clusters depend on intact actin filaments. Cholesterol depletion causes dispersion of syntaxin 3 but not syntaxin 4 clusters. In migrating cells, syntaxin clusters polarize to the leading edge, suggesting a role in polarized exocytosis. These results suggest that exocytosis occurs at small fusion sites exhibiting high local concentrations of SNARE proteins that may be required for efficient membrane fusion. The establishment of separate clusters for each syntaxin suggests that the plasma membrane is inherently polarized on an ultrastructural level even before the establishment of true cell polarity. PMID- 16339083 TI - Integration of local features to a global percept by neural coupling. AB - The integration of different visual attributes into the percept of a single global shape is a central aspect of object processing. In hierarchically organized stimuli with local and global levels, the attentional focus largely determines which level is processed. Here we tested the hypothesis that object processing during attention to the global aspect of the stimulus is characterized by an increased neural coupling between visual areas reflecting the integration of local features. In the present experiment, we used global letters that were constructed by smaller local letters, and a cue signaled which spatial level should be identified. On the local level, only 1 relevant letter was presented laterally in 1 visual hemifield. In contrast, the global letter extended into both hemifields, and the integration of information from both hemispheres was necessary to identify the global stimulus. Therefore, we expected an increased functional coupling between hemispheres during global processing. This hypothesis was investigated using electroencephalographic recordings and an analysis of phase locking and coherence. The results show that stimulus-locked neural coupling within the gamma band (30-40 Hz) across hemispheres in visual cortex increased for global processing after stimulus presentation and could therefore reflect the integration of local visual information. PMID- 16339082 TI - Depletion of phosphatidylcholine in yeast induces shortening and increased saturation of the lipid acyl chains: evidence for regulation of intrinsic membrane curvature in a eukaryote. AB - To study the consequences of depleting the major membrane phospholipid phosphatidylcholine (PC), exponentially growing cells of a yeast cho2opi3 double deletion mutant were transferred from medium containing choline to choline-free medium. Cell growth did not cease until the PC level had dropped below 2% of total phospholipids after four to five generations. Increasing contents of phosphatidylethanolamine (PE) and phosphatidylinositol made up for the loss of PC. During PC depletion, the remaining PC was subject to acyl chain remodeling with monounsaturated species replacing diunsaturated species, as shown by mass spectrometry. The remodeling of PC did not require turnover by the SPO14-encoded phospholipase D. The changes in the PC species profile were found to reflect an overall shift in the cellular acyl chain composition that exhibited a 40% increase in the ratio of C16 over C18 acyl chains, and a 10% increase in the degree of saturation. The shift was stronger in the phospholipid than in the neutral lipid fraction and strongest in the species profile of PE. The shortening and increased saturation of the PE acyl chains were shown to decrease the nonbilayer propensity of PE. The results point to a regulatory mechanism in yeast that maintains intrinsic membrane curvature in an optimal range. PMID- 16339084 TI - Domain specificity in visual cortex. AB - We investigated the prevalence and specificity of category-selective regions in human visual cortex. In the broadest survey to date of category selectivity in visual cortex, 12 participants were scanned with functional magnetic resonance imaging while viewing scenes and 19 different object categories in a blocked design experiment. As expected, we found selectivity for faces in the fusiform face area (FFA), for scenes in the parahippocampal place area (PPA), and for bodies in the extrastriate body area (EBA). In addition, we describe 3 main new findings. First, evidence for the selectivity of the FFA, PPA, and EBA was strengthened by the finding that each area responded significantly more strongly to its preferred category than to the next most effective of the remaining 19 stimulus categories tested. Second, a region in the middle temporal gyrus that has been reported to respond significantly more strongly to tools than to animals did not respond significantly more strongly to tools than to other nontool categories (such as fruits and vegetables), casting doubt on the characterization of this region as tool selective. Finally, we did not find any new regions in the occipitotemporal pathway that were strongly selective for other categories. Taken together, these results demonstrate both the strong selectivity of a small number of regions and the scarcity of such regions in visual cortex. PMID- 16339085 TI - Ambient GABA promotes cortical entry of tangentially migrating cells derived from the medial ganglionic eminence. AB - During corticogenesis, cells from the medial ganglionic eminence (MGE) migrate tangentially into the neocortical anlage. Here we report that gamma-aminobutyric acid (GABA), via GABAA receptors, regulates tangential migration. In embryonic telencephalic slices, bicuculline produced an outward current in migrating MGE derived cells in the neocortex, suggesting the presence of and tonic activation by ambient GABA. Ambient GABA was also present in the MGE, although this required demonstration using as bioassay HEK293 cells expressing high-affinity alpha6/beta2/gamma2s recombinant GABAA receptors. The concentration of ambient GABA was 0.5+/-0.1 microM in both regions. MGE-derived cells before the corticostriate juncture (CSJ) were less responsive to GABA than those in the neocortex, and profiling of GABAA receptor subunit transcripts revealed different expression patterns in the MGE vis-a-vis the neocortex. These findings suggest a dynamic expression of GABAA receptor number or isoform as MGE-derived cells enter the neocortex and become tonically influenced by ambient GABA. Treatment with bicuculline or antibody against GABA did not affect migration of MGE-derived cells before the CSJ but decreased "crossing index," reflecting impeded migration past the CSJ into the neocortex. Treatment with diazepam or addition of exogenous GABA increased crossing index. We conclude that ambient GABA promotes cortical entry of tangentially migrating MGE-derived cells. PMID- 16339086 TI - Repetitive transcranial magnetic stimulation effects on brain function and cognition among elders with memory dysfunction. A randomized sham-controlled study. AB - In the present study, we aimed to investigate the effects of repetitive transcranial magnetic stimulation (rTMS) on memory performance and brain activity in elders presenting with subjective memory complaints and a memory performance within the low normal range. Forty participants underwent 2 functional magnetic resonance imaging (fMRI) sessions, in which they were administered 2 equivalent face-name memory tasks. Following each fMRI, subjects were asked to pair faces with their corresponding proper name. In-between, high-frequency rTMS was applied randomly using real or sham stimulation in a double-blind design. Only subjects who received active rTMS improved in associative memory significantly. This was accompanied by additional recruitment of right prefrontal and bilaterial posterior cortical regions at the second fMRI session, relative to baseline scanning. Our findings reflect a potentiality of rTMS to recruit compensatory networks, which participate during the memory-encoding process. Present results represent the first evidence that rTMS is capable of transitorily and positively influencing brain function and cognition among elders with memory complaints. PMID- 16339087 TI - Cortical area MSTd combines visual cues to represent 3-D self-movement. AB - As arboreal primates move through the jungle, they are immersed in visual motion that they must distinguish from the movement of predators and prey. We recorded dorsal medial superior temporal (MSTd) cortical neuronal responses to visual motion stimuli simulating self-movement and object motion. MSTd neurons encode the heading of simulated self-movement in three-dimensional (3-D) space. 3-D heading responses can be evoked either by the large patterns of visual motion in optic flow or by the visual object motion seen when an observer passes an earth fixed landmark. Responses to naturalistically combined optic flow and object motion depend on their relative directions: an object moving as part of the optic flow field has little effect on neuronal responses. In contrast, an object moving separately from the optic flow field has large effects, decreasing the amplitude of the population response and shifting the population's heading estimate to match the direction of object motion as the object moves toward central vision. These effects parallel those seen in human heading perception with minimal effects of objects moving with the optic flow and substantial effects of objects violating the optic flow. We conclude that MSTd can contribute to navigation by supporting 3-D heading estimation, potentially switching from optic flow to object cues when a moving object passes in front of the observer. PMID- 16339088 TI - Cortical sources of CRF, NKB, and CCK and their effects on pyramidal cells in the neocortex. AB - In order to investigate how neuropeptide transmission can modulate the neocortical network, we mapped the expression of neurokinin (NK) B, cholecystokinin (CCK), and corticotropin-releasing factor (CRF) and their receptors to neuronal types using patch-clamp and single-cell reverse transcription-polymerase chain reaction in acute slices of rat neocortex. Classification of neurons by unsupervised clustering based on the analysis of multiple electrophysiological and molecular properties disclosed 3 GABAergic interneuron clusters and 1 pyramidal cell cluster. The 3 neuropeptides were expressed in a cluster of interneurons characteristically expressing vasoactive intestinal peptide. CRF was additionally found in a cluster containing almost exclusively somatostatin-expressing interneurons, whereas CCK was present in all clusters. The respective receptors of these peptides, NK-3, CCK-B, and CRF-1, were essentially expressed in pyramidal cells. At -60 mV, pyramidal cells were weakly depolarized by each of these peptides. When pyramidal neurons were maintained to about 5 mV below spike threshold, depolarization induced by each peptide resulted in a long-lasting action potential discharge. Neuropeptide effects were prevented by selective antagonists of NK-3, CCK-B, and CRF-1 receptors. These results suggest that pyramidal neurons are the primary target of NKB, CCK, and CRF in the neocortex. They further indicate that specific interneuron types coordinate the release of these peptides and can induce a long lasting increase of the excitability of the neocortical network. PMID- 16339089 TI - Images in clinical medicine. Deforming gout. PMID- 16339090 TI - Clinician-discoverers--Marshall, Warren, and H. pylori. PMID- 16339091 TI - Robert Koch, the Nobel Prize, and the ongoing threat of tuberculosis. PMID- 16339092 TI - The medical detectives. PMID- 16339093 TI - Remembering Berton Roueche--master of medical mysteries. PMID- 16339094 TI - Febuxostat compared with allopurinol in patients with hyperuricemia and gout. AB - BACKGROUND: Febuxostat, a novel nonpurine selective inhibitor of xanthine oxidase, is a potential alternative to allopurinol for patients with hyperuricemia and gout. METHODS: We randomly assigned 762 patients with gout and with serum urate concentrations of at least 8.0 mg per deciliter (480 micromol per liter) to receive either febuxostat (80 mg or 120 mg) or allopurinol (300 mg) once daily for 52 weeks; 760 received the study drug. Prophylaxis against gout flares with naproxen or colchicine was provided during weeks 1 through 8. The primary end point was a serum urate concentration of less than 6.0 mg per deciliter (360 micromol per liter) at the last three monthly measurements. The secondary end points included reduction in the incidence of gout flares and in tophus area. RESULTS: The primary end point was reached in 53 percent of patients receiving 80 mg of febuxostat, 62 percent of those receiving 120 mg of febuxostat, and 21 percent of those receiving allopurinol (P<0.001 for the comparison of each febuxostat group with the allopurinol group). Although the incidence of gout flares diminished with continued treatment, the overall incidence during weeks 9 through 52 was similar in all groups: 64 percent of patients receiving 80 mg of febuxostat, 70 percent of those receiving 120 mg of febuxostat, and 64 percent of those receiving allopurinol (P=0.99 for 80 mg of febuxostat vs. allopurinol; P=0.23 for 120 mg of febuxostat vs. allopurinol). The median reduction in tophus area was 83 percent in patients receiving 80 mg of febuxostat and 66 percent in those receiving 120 mg of febuxostat, as compared with 50 percent in those receiving allopurinol (P=0.08 for 80 mg of febuxostat vs. allopurinol; P=0.16 for 120 mg of febuxostat vs. allopurinol). More patients in the high-dose febuxostat group than in the allopurinol group (P=0.003) or the low-dose febuxostat group discontinued the study. Four of the 507 patients in the two febuxostat groups (0.8 percent) and none of the 253 patients in the allopurinol group died; all deaths were from causes that the investigators (while still blinded to treatment) judged to be unrelated to the study drugs (P=0.31 for the comparison between the combined febuxostat groups and the allopurinol group). CONCLUSIONS: Febuxostat, at a daily dose of 80 mg or 120 mg, was more effective than allopurinol at the commonly used fixed daily dose of 300 mg in lowering serum urate. Similar reductions in gout flares and tophus area occurred in all treatment groups. PMID- 16339095 TI - Infliximab for induction and maintenance therapy for ulcerative colitis. AB - BACKGROUND: Infliximab, a chimeric monoclonal antibody directed against tumor necrosis factor alpha, is an established treatment for Crohn's disease but not ulcerative colitis. METHODS: Two randomized, double-blind, placebo-controlled studies--the Active Ulcerative Colitis Trials 1 and 2 (ACT 1 and ACT 2, respectively)--evaluated the efficacy of infliximab for induction and maintenance therapy in adults with ulcerative colitis. In each study, 364 patients with moderate-to-severe active ulcerative colitis despite treatment with concurrent medications received placebo or infliximab (5 mg or 10 mg per kilogram of body weight) intravenously at weeks 0, 2, and 6 and then every eight weeks through week 46 (in ACT 1) or week 22 (in ACT 2). Patients were followed for 54 weeks in ACT 1 and 30 weeks in ACT 2. RESULTS: In ACT 1, 69 percent of patients who received 5 mg of infliximab and 61 percent of those who received 10 mg had a clinical response at week 8, as compared with 37 percent of those who received placebo (P<0.001 for both comparisons with placebo). A response was defined as a decrease in the Mayo score of at least 3 points and at least 30 percent, with an accompanying decrease in the subscore for rectal bleeding of at least 1 point or an absolute rectal-bleeding subscore of 0 or 1. In ACT 2, 64 percent of patients who received 5 mg of infliximab and 69 percent of those who received 10 mg had a clinical response at week 8, as compared with 29 percent of those who received placebo (P<0.001 for both comparisons with placebo). In both studies, patients who received infliximab were more likely to have a clinical response at week 30 (P< or =0.002 for all comparisons). In ACT 1, more patients who received 5 mg or 10 mg of infliximab had a clinical response at week 54 (45 percent and 44 percent, respectively) than did those who received placebo (20 percent, P<0.001 for both comparisons). CONCLUSIONS: Patients with moderate-to-severe active ulcerative colitis treated with infliximab at weeks 0, 2, and 6 and every eight weeks thereafter were more likely to have a clinical response at weeks 8, 30, and 54 than were those receiving placebo. (ClinicalTrials.gov numbers, NCT00036439 and NCT00096655.) PMID- 16339096 TI - Renal-cell carcinoma. PMID- 16339097 TI - Images in clinical medicine. Pseudomembranous colitis associated with Clostridium difficile. PMID- 16339098 TI - Case records of the Massachusetts General Hospital. Case 37-2005. A 35-year-old man with cardiac arrest while sleeping. PMID- 16339099 TI - Febuxostat--treatment for hyperuricemia and gout? PMID- 16339100 TI - Understanding heme transport. PMID- 16339101 TI - Fluid solutions in dengue shock syndrome. PMID- 16339102 TI - Dengue in travelers. PMID- 16339103 TI - Perioperative beta-blocker therapy and mortality. PMID- 16339104 TI - Dextrocardia with situs inversus. PMID- 16339105 TI - Secret insulin-injection syndrome among adolescents with type 1 diabetes. PMID- 16339106 TI - Case 28-2005: a case of systemic necrotizing vasculitis. PMID- 16339107 TI - Successful pregnancy after orthotopic liver transplantation in a patient with HIV infection. PMID- 16339108 TI - Tapping into renewal. PMID- 16339109 TI - Living Longer and Paying the Price? AB - Over the past century, there have been tremendous increases in longevity in the United States and most other developed countries. If these trends continue, the costs of paying for public programs like Social Security, Medicare, and Medicaid will escalate at a startling rate with the aging of the "baby boomer" generation. A meeting titled "Living Longer and Paying the Price?" was organized to consider whether current trends in longevity will continue at the current pace, accelerate, or decelerate as a result of public health problems such as obesity and infectious diseases. Further, speakers presented their views on covering the costs of public and private programs for future generations of older adults. PMID- 16339110 TI - Thematic review series: lipid posttranslational modifications. Fighting parasitic disease by blocking protein farnesylation. AB - Protein farnesylation is a form of posttranslational modification that occurs in most, if not all, eukaryotic cells. Inhibitors of protein farnesyltransferase (PFTIs) have been developed as anticancer chemotherapeutic agents. Using the knowledge gained from the development of PFTIs for the treatment of cancer, researchers are currently investigating the use of PFTIs for the treatment of eukaryotic pathogens. This "piggy-back" approach not only accelerates the development of a chemotherapeutic agent for protozoan pathogens but is also a means of mitigating the costs associated with de novo drug design. PFTIs have already been shown to be efficacious in the treatment of eukaryotic pathogens in animal models, including both Trypanosoma brucei, the causative agent of African sleeping sickness, and Plasmodium falciparum, one of the causative agents of malaria. Here, current evidence and progress are summarized that support the targeting of protein farnesyltransferase for the treatment of parasitic diseases. PMID- 16339111 TI - Sphingosylphosphorylcholine induces proliferation of human adipose tissue-derived mesenchymal stem cells via activation of JNK. AB - Sphingosylphosphorylcholine (SPC) has been implicated in a variety of cellular responses, including proliferation and differentiation. In this study, we demonstrate that d-erythro-SPC, but not l-threo-SPC, stereoselectively stimulated the proliferation of human adipose tissue-derived mesenchymal stem cells (hADSCs), with a maximal increase at 5 microM, and increased the intracellular concentration of Ca(2+) ([Ca(2+)](i)) in hADSCs, which do not express known SPC receptors (i.e., OGR1, GPR4, G2A, and GPR12). The SPC-induced proliferation and increase in [Ca(2+)](i) were sensitive to pertussis toxin (PTX) and the phospholipase C (PLC) inhibitor U73122, suggesting that PTX-sensitive G proteins, Gi or Go, and PLC are involved in SPC-induced proliferation. In addition, SPC treatment induced the phosphorylation of c-Jun and extracellular signal-regulated kinase, and SPC-induced proliferation was completely prevented by pretreatment with the c-Jun N-terminal kinase (JNK)-specific inhibitor SP600125 but not with the MEK-specific inhibitor U0126. Furthermore, the SPC-induced proliferation and JNK activation were completely attenuated by overexpression of a dominant negative mutant of JNK2, and the SPC-induced activation of JNK was inhibited by pretreatment with PTX or U73122. Treatment of hADSCs with lysophosphatidic acid (LPA) receptor antagonist, Ki16425, had no impact on the SPC-induced increase in [Ca(2+)](i). However, SPC-induced proliferation was partially, but significantly, attenuated by pretreatment of the cells with Ki16425.These results indicate that SPC stimulates the proliferation of hADSCs through the Gi/Go-PLC-JNK pathway and that LPA receptors may be responsible in part for the SPC-induced proliferation. PMID- 16339112 TI - Hepatic SR-BI-mediated cholesteryl ester selective uptake occurs with unaltered efficiency in the absence of cellular energy. AB - Scavenger receptor class B type I (SR-BI) plays a critical role in the delivery of HDL cholesterol and cholesteryl esters (CEs) to liver and steroidogenic tissues by a selective process that does not result in significant degradation of HDL protein. Recently, SR-BI-mediated endocytosis and recycling of HDL have been demonstrated. However, it remains unclear whether efficient SR-BI-mediated selective uptake occurs strictly at the plasma membrane or at additional sites along its endocytic itinerary. To examine the requirement for SR-BI endocytosis in HDL selective uptake, we determined the effects of energy depletion on the levels of cell-associated HDL protein and CE in primary mouse hepatocytes. Compared with CHO cells, we observed a much larger energy-dependent effect on CE uptake in primary mouse hepatocytes. Although varying the levels of caveolin-1 and carboxyl ester lipase altered the efficiency of selective uptake, neither was able to account for the energy-dependent component of HDL-CE uptake. Finally, we demonstrate that the hepatocyte-specific, energy-dependent effects on HDL apolipoprotein A-I and -CE uptake are independent of SR-BI and are not required to achieve efficient SR-BI-mediated selective uptake of CE. Together, these data support the conclusion that neither the intracellular trafficking of HDL nor any energy-dependent cellular process affects the ability of the cell to maximally acquire CE through SR-BI-mediated selective uptake from HDL. PMID- 16339113 TI - LDL receptor deficiency or apoE mutations prevent remnant clearance and induce hypertriglyceridemia in mice. AB - We have used adenovirus-mediated gene transfer and bolus injection of purified apolipoprotein E (apoE) in mice to determine the contribution of LDL receptor family members in the clearance of apoE-containing lipoproteins in vivo and the factors that trigger hypertriglyceridemia. A low dose [5 x 10(8) plaque-forming units (pfu)] of an adenovirus expressing apoE4 did not normalize plasma cholesterol levels of apolipoprotein E-deficient (apoE(-/-)) x low density lipoprotein receptor-deficient (LDLr(-/-)) mice and induced hypertriglyceridemia. A similar phenotype of combined dyslipidemia was induced in apoE(-/-) or apoE(-/ ) x LDLr(-/-) mice after infection with a low dose (4 x 10(8) pfu) of an adenovirus expressing the apoE4[R142V/R145V] mutant previously shown to be defective in receptor binding. In contrast, a low dose of 5 x 10(8) pfu of the apoE4-expressing adenovirus corrected hypercholesterolemia in apoE(-/-) mice and did not trigger hypertriglyceridemia. Bolus injection of purified apoE in apoE(-/ ) x LDLr(-/-) mice did not clear plasma cholesterol levels and induced mild hypertriglyceridemia. In contrast, similar injection of apoE in apoE(-/-) mice cleared plasma cholesterol and caused transiently mild hypertriglyceridemia. These findings suggest that a) the LDL receptor alone can account for the clearance of apoE-containing lipoproteins in mice, and the contribution of other receptors is minimal, and b) defects in either the LDL receptor or in apoE that affect its interactions with the LDL receptor, increase the sensitivity to apoE induced hypertriglyceridemia in mice. PMID- 16339114 TI - Improved alignment of nucleosome DNA sequences using a mixture model. AB - DNA sequences that are present in nucleosomes have a preferential approximately 10 bp periodicity of certain dinucleotide signals, but the overall sequence similarity of the nucleosomal DNA is weak, and traditional multiple sequence alignment tools fail to yield meaningful alignments. We develop a mixture model that characterizes the known dinucleotide periodicity probabilistically to improve the alignment of nucleosomal DNAs. We assume that a periodic dinucleotide signal of any type emits according to a probability distribution around a series of 'hot spots' that are equally spaced along nucleosomal DNA with 10 bp period, but with a 1 bp phase shift across the middle of the nucleosome. We model the three statistically most significant dinucleotide signals, AA/TT, GC and TA, simultaneously, while allowing phase shifts between the signals. The alignment is obtained by maximizing the likelihood of both Watson and Crick strands simultaneously. The resulting alignment of 177 chicken nucleosomal DNA sequences revealed that all 10 distinct dinucleotides are periodic, however, with only two distinct phases and varying intensity. By Fourier analysis, we show that our new alignment has enhanced periodicity and sequence identity compared with center alignment. The significance of the nucleosomal DNA sequence alignment is evaluated by comparing it with that obtained using the same model on non nucleosomal sequences. PMID- 16339115 TI - Obesity in older adults: technical review and position statement of the American Society for Nutrition and NAASO, The Obesity Society. AB - Obesity causes serious medical complications and impairs quality of life. Moreover, in older persons, obesity can exacerbate the age-related decline in physical function and lead to frailty. However, appropriate treatment for obesity in older persons is controversial because of the reduction in relative health risks associated with increasing body mass index and the concern that weight loss could have potential harmful effects in the older population. This joint position statement from the American Society for Nutrition and NAASO, The Obesity Society reviews the clinical issues related to obesity in older persons and provides health professionals with appropriate weight-management guidelines for obese older patients. The current data show that weight-loss therapy improves physical function, quality of life, and the medical complications associated with obesity in older persons. Therefore, weight-loss therapy that minimizes muscle and bone losses is recommended for older persons who are obese and who have functional impairments or medical complications that can benefit from weight loss. PMID- 16339116 TI - Diacylglycerol: efficacy and mechanism of action of an anti-obesity agent. AB - Obesity is at the forefront of global health issues and directly contributes to many chronic illnesses. Several dietary components show promise in the treatment of obesity, one of which is oil rich in diacylglycerols (DAGs). Present objectives are to examine scientific knowledge concerning DAG to assess evidence supporting the effects on substrate oxidation rates, body weight and fat mass, and blood lipids, and to assess safety, as well as elucidate potential mechanisms of action. DAG can be synthesized by an enzymatic process to produce mainly 1,3 isoform DAG. This 1,3-DAG oil is believed to have the ability to increase beta oxidation, to enhance body weight loss, to suppress body fat accumulation, and to lower serum triacylglycerol levels postprandially. While certain animal and human studies indicate that consumption of 1,3-DAG has positive physiological effects, others report no effect. The mechanisms of action of DAG are suggested to decrease the resynthesis of chylomicrons as well as shunting them directly to the liver through the portal vein, where they are oxidized. This increased fat oxidation may influence control of food intake by increasing satiety. Further study into the precise mechanism is required to understand its effects. Safety studies show no risks in consuming a diet rich in DAG oil. Overall, consumption of oils with higher amounts of DAG, specifically 1,3-DAG, may be useful in the battle against obesity. PMID- 16339118 TI - Quantitative trait loci for metabolic syndrome in the Hypertension Genetic Epidemiology Network study. AB - As part of the Hypertension Genetic Epidemiology Network study, genome scans were performed in two ethnicities on the categorical metabolic syndrome (MetS). Genome scans were performed also on the factor scores produced by factor analysis (quantitative MetS). Heritabilities were highest for the obesity-insulin (INS) factor and lowest for blood pressure (BP) and central obesity. Seventeen unique putative quantitative trait loci (QTLs) yielded logarithm of the odds ratio (LOD) scores in excess of 1.7, 8 for blacks and 9 for whites. Important QTL findings in whites included an LOD score of 3.19 on chromosome 15q15 for the BP factor, 3.08 on chromosome 8p23 for the lipids-INS factor, and 3.07 on chromosome 3p26 for the obesity-INS factor. In blacks, after excluding type 2 diabetics, important QTLs were identified, including an LOD score of 2.77 on 13p12 for the obesity-INS factor and 2.63 on chromosome 11q24 for the lipids-INS factor. Categorical MetS had lower results than quantitative MetS. Notably, several loci identified overlap with those identified in other studies for a single or group of traits. The most promising candidate loci on 11q24 for lipids-INS and 13p12 for obesity INS in blacks, 8p23 for lipids-INS, 14q24 for obesity-INS, and 15q15 for BP in whites warrant further investigation. PMID- 16339117 TI - Linkage analysis of factors underlying insulin resistance: Strong Heart Family Study. AB - In previous work in non-diabetic participants of the Strong Heart Family Study, we identified three heritable principal components of nine insulin resistance (IR) phenotypes: 1) a glucose/insulin/obesity factor, 2) a blood pressure factor, and 3) a dyslipidemia factor. To localize quantitative trait loci (QTL) potentially influencing these factors, we conducted a genome scan of factor scores in Strong Heart Family Study participants. Approximately 599 men and women, >or=18 years of age, in 32 extended families at three centers (in Arizona, Oklahoma, and North and South Dakota), were examined between 1997 and 1999. We used variance components linkage analysis to identify QTLs for the IR factors. With age, sex, and study center as covariates, we detected linkage of the glucose/insulin/obesity factor to chromosome 4 (robust logarithm of the odds (LOD) = 2.2), the dyslipidemia factor to chromosome 12 (robust LOD = 2.7), and the blood pressure factor to chromosome 1 (robust LOD = 1.6). The peak linkage signals identified for these IR factors support several positive findings from other studies and occur in regions harboring interesting candidate genes. The corroboration of existing QTLs will bring us closer to the identification of the functional genes that predispose to IR. PMID- 16339119 TI - Quantitative trait loci for lipid content in Drosophila melanogaster. AB - Recombinant inbred lines derived from a natural population were used to investigate natural genetic variation for lipid abundance, protein abundance, and weight of Drosophila melanogaster. Females were heavier and contained more lipid and soluble protein than males. Lipid and protein abundance were genetically correlated with female weight, but male weight was not correlated with lipid or protein. Lipid and protein abundance were genetically correlated in males, but not in females. Quantitative trait loci (QTLs) for weight and protein abundance were predominantly on the X chromosome, whereas QTLs for lipid abundance were found on the second and third chromosomes. QTLs for lipid proportion (lipid abundance normalized by weight or protein abundance) were present on all chromosomes; a lipid proportion QTL on the third chromosome correlated with a QTL for starvation resistance observed in a previous study using the same set of recombinant inbred lines, suggesting that it might underlie both traits. Candidate genes are discussed in relationship to lipid abundance, lipid proportion, and starvation resistance. PMID- 16339120 TI - Pattern profiling and mapping of the fat body transcriptome in Drosophila melanogaster. AB - In Drosophila, the fat body is a collective name for the masses and sheets of adipose tissue that are distributed throughout the fly body. Thus far, >386,000 Drosophila expressed sequence tags (ESTs) have been deposited to the GenBank database, including 10,443 derived from fat body in flies (data accessed on October 7, 2004). The objective of this study was to map the transcriptome of the fat body in flies and thus provide genomics and bioinformatics tools for developing a Drosophila model for addressing the genetic complexity of obesity in humans. The gene-EST Basic Local Alignment Search Tool (BLAST) matches revealed that these ESTs could represent 12,188 coding genes in the Drosophila genome. Among them, at least 2,261 are expressed in the fat body, including 41 identified as preferentially expressed genes with logarithm of odds >3.0. Self-organizing map analysis revealed a cluster of 290 genes favorably expressed in the fat body compared with genes expressed in five other tissues. Mapping of the fat body transcriptome identified a 1.7-Mb domain on 3L containing 35 genes that were expressed at a much higher level than in other tissues (transcript density factor = 1.0 approximately 2.3). PMID- 16339121 TI - Trends in obesity and overweight-related office visits and drug prescriptions in Canada, 1998 to 2004. AB - OBJECTIVE: Obesity and overweight are affecting increasing numbers of Canadians and have received considerable amounts of medical, governmental, and media attention in recent years. This study sought to determine whether this rise in prevalence and awareness has resulted in an increased frequency of obesity and overweight-related office visits or antiobesity drug prescriptions over the past 5 years. RESEARCH METHODS AND PROCEDURES: Data from IMS Health Canada were used to derive nationally representative estimates of trends in the annual number of obesity and overweight-related office visits (1999 to 2003) and the quarterly prescription volume of antiobesity drugs (July 1998 to March 2003) in Canada. RESULTS: The number of obesity and overweight-related office visits increased by 20% between 1999 and 2000 but then remained constant. The number of antiobesity drug prescriptions peaked in 2001 and has since declined, with parallel trends being observed for all individual agents. In contrast, the overall frequency of office visits and drug prescriptions in Canada (for any reason) progressively increased over the study period. Middle-aged women were the most common type of patient to seek physician advice regarding obesity, and general practitioners were the most common type of physician visited. DISCUSSION: Increases in the prevalence and awareness of obesity have not resulted in major increases in office visits or drug prescriptions for this condition over the past 5 years. A number of patient, physician, and drug-related factors may explain these results, which are likely a reflection primarily of the current lack of effective weight loss strategies for obese individuals. PMID- 16339122 TI - Effects of irbesartan on the growth and differentiation of adipocytes in obese zucker rats. AB - OBJECTIVE: The aim of this study was to evaluate the effects of the selective angiotensin receptor 1 antagonist irbesartan on the growth and differentiation of the adipocytes in obese Zucker fa/fa rats. RESEARCH METHODS AND PROCEDURES: Obese Zucker fa/fa rats were treated by oral route for 3 weeks with irbesartan at doses of 3-10-30 mg/kg per day. The adipocyte differentiation was evaluated by analyzing tissue samples of white (retroperitoneal) or brown (interscapular) adipose tissue for the presence of peroxisome proliferator activated receptor gamma, leptin, and the activity of glycerol-3-phosphate dehydrogenase. RESULTS: This study showed that the treatment of obese Zucker fa/fa with irbesartan effectively reduced the differentiation of adipocytes within brown (interscapular) and white (retroperitoneal) adipose tissue. In fact, irbesartan significantly (p < 0.01) and dose-dependently reduced the tissue levels of leptin, peroxisome proliferator activated receptor gamma, and the activity of the enzyme glycerol-3-phoshate dehydrogenase accepted markers of adipocyte differentiation. None of the tested doses of irbesartan affected these markers in non-obese rats. DISCUSSION: The antagonism of the angiotensin receptor 1 receptors with irbesartan reduces the adipogenic activity of angiotensin II in obese Zucker rats, with the endpoint being reduction of the growth and differentiation of the adipocytes within the adipose tissue. PMID- 16339123 TI - (+)-Z-Bisdehydrodoisynolic acid ameliorates obesity and the metabolic syndrome in female ZDF rats. AB - OBJECTIVE: The putative selective estrogen receptor modulator (+)-Z bisdehydrodoisynolic acid (Z-BDDA) has been found to improve cardiovascular risk in rodents. The objective of this study was to investigate the effectiveness of (+)-Z-BDDA compared with the antidiabetic drug, rosiglitazone, in treating obesity and risk factors associated with the metabolic syndrome. RESEARCH METHODS AND PROCEDURES: Female Zucker Diabetic Fatty rats were randomly assigned to three treatment groups for 29 weeks: control (C), 1.8 mg (+)-Z-BDDA/kg diet [control diet + (+)-Z-BDDA (CB)], or 100 mg rosiglitazone/kg diet [control diet + rosiglitazone (CR)]. At sacrifice, physiological, biochemical, and molecular parameters were examined. RESULTS: CB animals gained less weight and exhibited a decrease in total body lipids (p < 0.05) as compared with C or CR rats. Body weight and total body lipids were the highest in CR rats (p < 0.05). Liver weights in CB and CR rats were lower (p < 0.05) than in C rats, whereas kidney weights were lower in CB (p < 0.05) than in C and CR animals. Fasting plasma glucose was lower (p < 0.05) in the CB and CR animals when compared with C animals. C rats exhibited the highest concentration of total plasma cholesterol, and CR-treated rats exhibited the lowest concentration. Plasma triglycerides followed the same pattern as plasma cholesterol. Histomorphometry of heart vasculature revealed that CB and CR treatments produced a significant shift from small to large venules and arterioles compared with C (p < 0.05). Liver expression profiles of peroxisome proliferator-activated receptor (PPAR) alpha, PPARgamma, and PPAR-regulated genes revealed encouraging CB-induced effects. DISCUSSION: These results suggest that (+)-Z-BDDA may have applications in treating obesity and complications associated with the metabolic syndrome. PMID- 16339124 TI - Potential role of interleukin-18 in liver disease associated with insulin resistance. AB - OBJECTIVE: Interleukin (IL)-18 has been associated with obesity and insulin resistance, both risk factors for the development of liver disease, but the role of IL-18 in liver disease associated with insulin resistance is presently unknown. We hypothesized that circulating IL-18 would be related to serum concentrations of liver chemistry tests (LCTs) in apparently healthy subjects and wished to study whether this correlation was dependent on insulin sensitivity (S(I)). RESEARCH METHODS AND PROCEDURES: One hundred six apparently healthy white men consecutively enrolled in a cross-sectional, population-based study dealing with S(I) in men were studied, and S(I) (minimal model analysis), LCTs (colorimetry), and IL-18 serum concentrations (immunoassay) were assessed. RESULTS: Compared with subjects in the lowest quartile for serum IL-18, subjects in the highest quartile exhibited increased serum triglycerides and decreased S(I), in addition to higher serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (all p < 0.05). The direct association between both ALT and AST and IL-18 was further confirmed by examining the distribution of serum IL 18 by quartiles of ALT and AST. Subjects in the highest quartile for serum ALT and AST had higher IL-18 concentrations compared with subjects in the lowest quartile for these LCTs (both p = 0.01). In multiple regression analysis, IL-18, but not S(I), was an independent predictor of serum concentrations of ALT and AST, explaining 7% and 4% of their variance, respectively. DISCUSSION: In summary, IL-18 serum concentrations are associated in apparently healthy humans with plasma concentrations of various LCTs. IL-18 could contribute to the development of liver disease associated with insulin resistance. PMID- 16339125 TI - Meal frequency and childhood obesity. AB - OBJECTIVE: Previous studies have demonstrated an inverse association between meal frequency and the prevalence of obesity in adulthood. The aim of this study was to assess the relationship between meal frequency and childhood obesity. RESEARCH METHODS AND PROCEDURES: Stature and weight of 4,370 German children ages 5 to 6 years were determined in six Bavarian (Germany) public health offices during the obligatory school entry health examination in 2001/2002. An extensive questionnaire on risk factors for obesity was answered by their parents. Obesity was defined according to sex- and age-specific BMI cut-off points proposed by the International Obesity Task Force. The main exposure was daily meal frequency. RESULTS: The prevalence of obesity decreased by number of daily meals: three or fewer meals, 4.2% [95% confidence interval (CI), 2.8 to 6.1]; four meals, 2.8% (95% CI, 2.1 to 3.7); and 5 or more meals, 1.7% (95% CI, 1.2 to 2.4). These effects could not be explained by confounding due to a wide range of constitutional, sociodemographic, and lifestyle factors. The adjusted odds ratios for obesity were 0.73 (95% CI, 0.44 to 1.21) for four meals and 0.51 (95% CI, 0.29 to 0.89) for five or more meals. Additional analyses pointed to a higher energy intake in nibblers compared with gorgers. DISCUSSION: A protective effect of an increased daily meal frequency on obesity in children was observed and appeared to be independent of other risk factors for childhood obesity. A modulation of the response of hormones such as insulin might be instrumental. PMID- 16339126 TI - Association of family environment with children's television viewing and with low level of physical activity. AB - OBJECTIVE: This study examined associations between the family environment and children's television (TV) viewing and likelihood of being low-active. RESEARCH METHODS AND PROCEDURES: In 2001, children were recruited from 19 primary schools in Melbourne, Australia. Parents completed a questionnaire about their child's TV viewing and the family environment. Children also completed a questionnaire and wore an accelerometer for 8 days. Movement counts were used to identify low active children (lowest quartile). Data were analyzed in May 2004. RESULTS: The sample consisted of 878 children (mean age = 11.5 +/- 0.6 yrs). Multiple logistic regression revealed that socioeconomic status [adjusted odds ratios (AOR) = 0.4 boys], frequency families watched TV together (AOR = 2.0 boys), mothers' (AOR = 1.8 boys; AOR = 2.5 girls) and fathers' (AOR = 2.6 boys; AOR = 2.8 girls) TV viewing, and rules prohibiting TV during mealtimes (AOR = 0.6 boys; AOR = 0.6 girls) related to children watching TV >or=2 h/d. Variables associated with low level physical activity included self-reported enjoyment of Internet use (AOR = 1.7 boys) and preference for watching TV (AOR = 2.3 girls), perception that mother uses computer a lot (AOR = 1.9 boys) and likes using the computer (AOR = 0.6 girls), fathers' reported computer/electronic games use (AOR = 1.7 girls), frequency families used computer together (AOR = 0.4 girls), rules that TV viewing must be supervised (AOR = 1.9 boys; AOR = 0.6 girls), and having pay TV (AOR = 0.6 boys) and electronic games at home (AOR = 2.6 boys). DISCUSSION: These findings suggest that the relationships between the family environment and TV viewing and low-level activity are complex and that these behaviors are distinct. PMID- 16339128 TI - Weight loss expectations in obese patients and treatment attrition: an observational multicenter study. AB - OBJECTIVE: To investigate the influence of weight loss expectations (expected 1 year BMI loss, dream and maximum acceptable BMI) on attrition in obese patients seeking treatment. RESEARCH METHODS AND PROCEDURES: Obese subjects (1,785; 1,393 women; median age, 46 years; median BMI, 36.7 kg/m(2)) seeking treatment in 23 medical Italian centers were evaluated. Baseline diet and weight history, weight loss expectations, and primary motivation for seeking treatment (health or improving appearance) were systematically recorded. Psychiatric distress, binge eating, and body image dissatisfaction were tested at baseline by self administered questionnaires (Symptom Check List-90, Binge Eating Scale, and Body Uneasiness Test). Attrition and BMI change at 12 months were prospectively recorded. RESULTS: At 12 months, 923 of 1,785 patients (51.7%) had discontinued treatment. Compared with continuers, drop-outs had a significantly lower age, a lower age at first dieting, lower dream BMI, a higher expected 1-year BMI loss, and a higher weight phobia. At logistic regression analysis, the strongest predictors of attrition at 12 months were lower age and higher expected 1-year BMI loss. The risk of drop-out increased systematically for unit increase in expected BMI loss at 12 months (hazard ratio, 1.12; 95% confidence interval, 1.04 to 1.20; p = 0.0018). The risk was particularly elevated in the first 6 months. DISCUSSION: Baseline weight loss expectations are independent cognitive predictors of attrition in obese patients entering a weight-losing program; the higher the expectations, the higher attrition at 12 months. Unrealistic weight goals should be tackled at the very beginning of treatment. PMID- 16339127 TI - Dietary intake of whole and refined grain breakfast cereals and weight gain in men. AB - OBJECTIVE: Prospective studies have suggested that substituting whole grain for refined grain products may lower the risk of overweight and obesity. Breakfast cereal intake is a major source of whole and refined grains and has also been associated with having a lower BMI. The aim of this study was to prospectively assess the association between whole and refined grain breakfast cereal intakes and risk of overweight (BMI >or= 25 kg/m(2)) and weight gain. RESEARCH METHODS AND PROCEDURES: We examined 17,881 U.S. male physicians 40 to 84 years of age in 1982 who were free of cardiovascular disease, diabetes mellitus, and cancer at baseline and reported measures of breakfast cereal intake, weight, and height. RESULTS: Over 8 and 13 years of follow-up, respectively, men who consumed breakfast cereal, regardless of type, consistently weighed less than those who consumed breakfast cereals less often (p value for trend = 0.01). Whole and refined grain breakfast cereal intake was inversely associated with body weight gain over 8 years, after adjustment for age, smoking, baseline BMI, alcohol intake, physical activity, hypertension, high cholesterol, and use of multivitamins. Compared with men who rarely or never consumed breakfast cereals, those who consumed >or=1 serving/d of breakfast cereals were 22% and 12% less likely to become overweight during follow-up periods of 8 and 13 years (relative risk, 0.78 and 0.88; 95% confidence interval, 0.67 to 0.91 and 0.76 to 1.00, respectively). DISCUSSION: BMI and weight gain were inversely associated with intake of breakfast cereals, independently of other risk factors. PMID- 16339129 TI - Intergenerational perceptions of body image in hispanics: role of BMI, gender, and acculturation. AB - OBJECTIVE: To assess role of BMI, gender, and acculturation on maternal and children's perception of body size, body ideal, and attractiveness. RESEARCH METHODS AND PROCEDURES: Eighty mothers and their 6- to- 12-year-old children (41 boys, 39 girls) participated. Maternal and children's perceptions of body size (actual and ideal) and attractiveness were assessed through a pictorial instrument. Mother and child height and weight, demographic, and acculturation characteristics were also assessed. RESULTS: Seventy-nine percent of the mothers were overweight, and 32% of the boys and 34% of the girls were overweight or at risk for overweight. BMI influenced the children's selection of perceived ideal size. Overweight or at-risk for overweight children were more likely to select thinner figures as the ideal size than non-overweight children. Gender and acculturation differences concerning children's perceptions of body size and attractiveness were also found. Girls perceived the obese figure as being less attractive than did the boys. More acculturated children were likely to select thinner figures as more attractive than their less acculturated counterparts. Maternal acculturation was associated positively with the girls' choice of thinner figures as an ideal body size, but not with the boys. Mothers viewed their daughters' actual body size and BMI as ideal, although 34% of the girls were at-risk for overweight. Mothers perceived average body size figures as more attractive for their sons. DISCUSSION: Findings from this study provide empirical data about the role of BMI, gender, acculturation, and familial influences on children's perceptions of actual and ideal body sizes and attractiveness. PMID- 16339130 TI - Reexamining obesigenic families: parents' obesity-related behaviors predict girls' change in BMI. AB - OBJECTIVE: It has been shown that girls from families in which mothers and fathers had high dietary intake and low physical activity (i.e., obesigenic families) were at increased risk of obesity from ages 5 to 7 years. This follow up study uses additional data collected when girls were 9 and 11 years old to examine whether girls from obesigenic families continued to show greater increases in BMI over time and reported unhealthy dietary and activity patterns. RESEARCH METHODS AND PROCEDURES: Families from the original cohort were reexamined when girls were 9 and 11 years of age. Parents' and girls' BMI, dietary intake, and physical activity and girls' percentage body fat and television viewing were assessed. RESULTS: In comparison with girls from non obesigenic families, girls from obesigenic families showed greater increases in BMI and BMI z score from ages 5 to 7 years that were maintained across ages 7 to 11 years. Furthermore, girls from obesigenic families had higher percentage body fat at ages 9 and 11 years. These results were independent of parents' BMI. Additional findings showed that girls from obesigenic families had diets higher in percentage fat and had higher levels of television viewing than girls from non obesigenic families. DISCUSSION: The environment that parents create, by way of their own dietary and physical activity behaviors, may have a lasting negative effect on children's weight trajectories and their emerging obesity risk behaviors, such as their dietary patterns. These findings further highlight the importance of the family in establishing children's obesity risk and the necessity of targeting parents of young children in obesity prevention efforts. PMID- 16339131 TI - Major depression predicts an increase in long-term body weight variability in young adults. AB - OBJECTIVE: To test the hypothesis that major depression predicts an increase in long-term body weight variability (BWV). RESEARCH METHODS AND PROCEDURES: This was a prospective community-based single-age cohort study of young adults (N = 591) followed between the ages of 19 and 40. Following initial screening, information was derived from six subsequent semistructured diagnostic interviews conducted by mental health professionals. Major depression was diagnosed on the basis of DSM criteria. BWV was defined as the root mean square error of a regression line fitted to each individual's BMI values over time. Multiple regression analysis was used to test the association between major depression and BWV while controlling for potentially confounding variables including antidepressant treatment, eating disorder symptoms, and physical activity. We used random effects models to determine the temporal relationship between repeated measures of major depression and body weight change. RESULTS: A highly significant positive association between major depression and BWV was found, whereas major depression was not associated with BMI level or BMI trend. Depression severity showed a dose-response-type relationship with the magnitude of BWV. After controlling for potentially confounding variables including antidepressant use, eating disorder symptoms, smoking, and physical activity, major depression remained a significant predictor of BWV (beta= 0.13, p < 0.001). Longitudinal analysis revealed a unidirectional association between major depression and a later increase in body weight change rate irrespective of antidepressant medication. DISCUSSION: Results from this study implicate depression as an important risk factor for increased BWV. Given increasing evidence for a link between major depression and both diabetes and cardiovascular disease, current results encourage further research on depression, BWV, and negative health outcomes. PMID- 16339132 TI - Is maternal psychopathology related to obesigenic feeding practices at 1 year? AB - OBJECTIVE: To investigate the relationship between maternal psychopathological symptomatology during pregnancy and at 6 and 12 months postnatally and maternal use of controlling and restrictive feeding practices at 1 year. RESEARCH METHODS AND PROCEDURES: Eighty-seven women completed a measure of psychological distress during pregnancy and at 6 and 12 months postpartum, and at 12 months postnatally these women reported their usage of controlling and restrictive feeding practices and were observed feeding their infants. RESULTS: General psychological distress, particularly anxious psychopathology, during pregnancy and at 6 and 12 months postnatally was significantly associated with maternal use of restrictive feeding practices at 1 year, even when controlling for length of breast-feeding and the infants' weights at 1 year. Contrary to expectations, depression and eating psychopathology as measured by the SCOFF eating disorder measure during pregnancy or at 6 or 12 months postnatally were not associated with the use of controlling or restrictive feeding practices at 1 year. DISCUSSION: These findings indicate that anxious maternal psychopathology may partially explain the development of maternal use of restriction when feeding. PMID- 16339133 TI - Correlates of persistent thinness in black and white young women. AB - OBJECTIVE: To examine health and psychosocial correlates of persistent thinness in black and white young adult women. RESEARCH METHODS AND PROCEDURES: 1,830 females (n = 988 black, n = 842 white) who participated in the National Heart, Lung, and Blood Institute Growth and Health Study were asked to indicate their current body size from a series of nine pictograms (1, emaciated to 9, obese). Persistent thinness was defined as having at least seven non-missing measurement points between ages 9 to 18 with a body size rating of <4 at all points. Generalized linear models were used to examine whether persistently thin women differed from comparison women on cardiovascular disease risk factors measured at age 18.5 and psychosocial variables measured at age 21.5. Prospective associations between psychological measures in childhood and persistent thinness through the course of adolescence were also examined. RESULTS: In the sample, 145 women (7.9%) met criteria for persistent thinness and 1,685 women (92.1%) were classified as not persistently thin. Persistently thin women had a later age of menarche, fewer weight concerns, and healthier eating attitudes, were less likely to have had a child, came from higher socioeconomic backgrounds, and had significantly lower blood pressure and higher high-density lipoprotein cholesterol than comparison women. Differences were not found on measures of depression or health services use. Girls with higher self-esteem in childhood were more likely to remain persistently thin throughout adolescence. DISCUSSION: Persistently thin women seem to be healthier on several indicators relative to comparison women, and race did not moderate these differences. PMID- 16339134 TI - Relationship between plasma adiponectin levels and metabolic risk profiles in Taiwanese children. AB - OBJECTIVE: Adiponectin, a novel adipokine with antiinflammatory and insulin sensitizing properties, has an important role in glucose metabolism and is negatively correlated with body fat amount in adults. The purpose of this study was to evaluate the association of plasma adiponectin level with metabolic risk profiles and insulin resistance status among Taiwanese children. RESEARCH METHODS AND PROCEDURES: We enrolled 1,248 children (608 boys and 640 girls) to ascertain their demographic, anthropometric, and cardiovascular risk factors distribution in Taipei. We measured plasma insulin, adiponectin, and leptin levels by radioimmunoassay (Linco Research Inc, St. Charles, MO). We calculated an insulin resistance index (IRI) using the Homeostasis Model Assessment model and also calculated an insulin resistance syndrome (IRS) summary score for each individual by adding the quartile ranks from the distribution of systolic blood pressure, serum triglyceride, high-density lipoprotein-cholesterol (HDL-C) (inverse), and insulin levels. RESULTS: In general, the boys had larger BMI, higher systolic blood pressure, serum total cholesterol, and triglyceride, and lower plasma leptin and adiponectin levels than girls. Plasma adiponectin levels were correlated negatively with BMI, leptin, insulin, IRI, and IRS summary score but positively correlated with HDL-C in both boys and girls. In multivariate regression analyses, adiponectin was negatively associated with insulin (girls only), IRI (girls only), and IRS score, and positively associated with HDL-C in both genders even after adjusting for age, BMI, plasma leptin level, and other potential confounders. DISCUSSION: These data suggest that plasma adiponectin levels were negatively associated with metabolic risk profiles that may have played a protective role in the development of insulin resistance among Taiwanese school children. PMID- 16339135 TI - Associations of maternal prenatal smoking with child adiposity and blood pressure. AB - OBJECTIVE: To examine the extent to which maternal prenatal smoking is associated with adiposity, central adiposity, and blood pressure in 3-year-old children. RESEARCH METHODS AND PROCEDURES: We studied 746 mother-child pairs in Project Viva, a prospective cohort study, and categorized mothers as never, early pregnancy, or former smokers. Main outcome measures were overweight (BMI for age and sex > 85th percentile), BMI z-score, sum of subscapular (SS) and triceps (TR) skinfolds, SS:TR skinfold ratio, and systolic blood pressure (SBP). RESULTS: One hundred sixty-one (22%) mothers quit smoking before pregnancy, 71 (10%) smoked in early pregnancy, and 514 (69%) never smoked. At age 3 years, 204 (27%) children were overweight. On multivariable analysis, compared with children of never smokers, children of early pregnancy smokers had an elevated risk for overweight [odds ratio (OR), 2.2; 95% confidence interval (CI), 1.2, 3.9] and higher BMI z score (0.30 units; 95% CI, 0.05, 0.55), SS + TR (2.0 mm; 95% CI, 0.9, 3.0), and SBP (2.4 mm Hg; 95% CI, -0.1, 4.9). Children of former smokers were not more overweight (BMI z-score, 0.02 units; 95% CI, -0.15, 0.19) but had higher SBP (1.5 mm Hg; 95% CI, -0.1, 3.2). We saw no relationship of smoking with central adiposity (SS:TR). DISCUSSION: Former and early pregnancy smokers had children with somewhat higher SBP, but only early pregnancy smokers had children who were more overweight. Mechanisms linking smoking with child adiposity and blood pressure may differ. A long-term impact of maternal smoking on offspring cardiovascular risk provides further reason to reduce smoking in women. PMID- 16339136 TI - Population-based study of trends, costs, and complications of weight loss surgeries from 1990 to 2002. AB - OBJECTIVE: To describe the trends, costs, and complications associated with weight loss surgery (WLS). RESEARCH METHODS AND PROCEDURES: Wisconsin inpatient hospital discharge data from 1990 to 2003 were used for analysis. A WLS case was defined as anyone with a WLS-related procedure code and a primary diagnosis of morbid obesity. Charges were inflation-adjusted to 2001 constant dollars; complications were defined on the basis of readmission, extended length of stay, repeat surgical procedures, or death. RESULTS: The number of WLSs increased from 269 in 1990 to 1992 to 1,884 in 2000 to 2002 (rate ratio = 4.6). Increases in WLSs were greatest among those 50 to 59 years of age (rate ratio = 6.4), women (rate ratio = 6.8), and blacks (rate ratio = 20.0). Between the two periods, inflation-adjusted WLS charges increased 12-fold, and the inflation-adjusted charge per procedure doubled, despite a decreased length of stay. For 2000 to 2002, 23.3% of WLS patients had either an extended length of stay or readmission within 30 days, 7.4% required a repeat surgical procedure, and 0.7% died. DISCUSSION: In Wisconsin, the rate and costs of WLSs have increased dramatically, and the incidence of postoperative complications was high. The epidemic of obesity in the United States makes it imperative to better assess the cost effectiveness of WLS and to improve its safety. PMID- 16339137 TI - Ablation of mouse phosphomannose isomerase (Mpi) causes mannose 6-phosphate accumulation, toxicity, and embryonic lethality. AB - MPI encodes phosphomannose isomerase, which interconverts fructose 6-phosphate and mannose 6-phosphate (Man-6-P), used for glycoconjugate biosynthesis. MPI mutations in humans impair protein glycosylation causing congenital disorder of glycosylation Ib (CDG-Ib), but oral mannose supplements normalize glycosylation. To establish a mannose-responsive mouse model for CDG-Ib, we ablated Mpi and provided dams with mannose to rescue the anticipated defective glycosylation. Surprisingly, although glycosylation was normal, Mpi(-/-) embryos died around E11.5. Mannose supplementation even hastened their death, suggesting that man nose was toxic. Mpi(-/-) embryos showed growth retardation and placental hyperplasia. More than 90% of Mpi(-/-) embryos failed to form yolk sac vasculature, and 35% failed chorioallantoic fusion. We generated primary embryonic fibroblasts to investigate the mechanisms leading to embryonic lethality and found that mannose caused a concentration- and time-dependent accumulation of Man 6-P in Mpi(-/-) fibroblasts. In parallel, ATP decreased by more than 70% after 24 h compared with Mpi(+/+) controls. In cell lysates, Man-6 P inhibited hexokinase (70%), phosphoglucose isomerase (65%), and glucose-6 phosphate dehydrogenase (85%), but not phosphofructokinase. Incubating intact Mpi(-/-) fibroblasts with 2-[(3)H]deoxyglucose confirmed mannose-dependent hexokinase inhibition. Our results in vitro suggest that mannose toxicity in Mpi( /-) embryos is caused by Man-6-P accumulation, which inhibits glucose metabolism and depletes intracellular ATP. This was confirmed in E10.5 Mpi(-/-) embryos where Man-6-P increased more than 10 times, and ATP decreased by 50% compared with Mpi(+/+) littermates. Because Mpi ablation is embryonic lethal, a murine CDG Ib model will require hypomorphic Mpi alleles. PMID- 16339138 TI - Dominant role of thyrotropin-releasing hormone in the hypothalamic-pituitary thyroid axis. AB - Hypothalamic thyrotropin-releasing hormone (TRH) stimulates thyroid-stimulating hormone (TSH) secretion from the anterior pituitary. TSH then initiates thyroid hormone (TH) synthesis and release from the thyroid gland. Although opposing TRH and TH inputs regulate the hypothalamic-pituitary-thyroid axis, TH negative feedback is thought to be the primary regulator. This hypothesis, however, has yet to be proven in vivo. To elucidate the relative importance of TRH and TH in regulating the hypothalamic-pituitary-thyroid axis, we have generated mice that lack either TRH, the beta isoforms of TH receptors (TRbeta KO), or both (double KO). TRbeta knock-out (KO) mice have significantly higher TH and TSH levels compared with wild-type mice, in contrast to double KO mice, which have reduced TH and TSH levels. Unexpectedly, hypothyroid double KO mice also failed to mount a significant rise in serum TSH levels, and pituitary TSH immunostaining was markedly reduced compared with all other hypothyroid mouse genotypes. This impaired TSH response, however, was not due to a reduced number of pituitary thyrotrophs because thyrotroph cell number, as assessed by counting TSH immunopositive cells, was restored after chronic TRH treatment. Thus, TRH is absolutely required for both TSH and TH synthesis but is not necessary for thyrotroph cell development. PMID- 16339139 TI - Cell death in pancreatitis: caspases protect from necrotizing pancreatitis. AB - Mechanisms of cell death in pancreatitis remain unknown. Parenchymal necrosis is a major complication of pancreatitis; also, the severity of experimental pancreatitis correlates directly with necrosis and inversely with apoptosis. Thus, shifting death responses from necrosis to apoptosis may have a therapeutic value. To determine cell death pathways in pancreatitis and the possibility of necrosis/apoptosis switch, we utilized the differences between the rat model of cerulein pancreatitis, with relatively high apoptosis and low necrosis, and the mouse model, with little apoptosis and high necrosis. We found that caspases were greatly activated during cerulein pancreatitis in the rat but not mouse. Endogenous caspase inhibitor X-linked inhibitor of apoptosis protein (XIAP) underwent complete degradation in the rat but remained intact in the mouse model. Furthermore, XIAP inhibition with embelin triggered caspase activation in the mouse model, implicating XIAP in caspase blockade in pancreatitis. Caspase inhibitors decreased apoptosis and markedly stimulated necrosis in the rat model, worsening pancreatitis parameters. Conversely, caspase induction with embelin stimulated apoptosis and decreased necrosis in mouse model. Thus, caspases not only mediate apoptosis but also protect from necrosis in pancreatitis. One protective mechanism is through degradation of receptor-interacting protein (RIP), a key mediator of "programmed" necrosis. We found that RIP was cleaved (i.e. inactivated) in the rat but not the mouse model. Caspase inhibition restored RIP levels; conversely, caspase induction with embelin triggered RIP cleavage. Our results indicate key roles for caspases, XIAP, and RIP in the regulation of cell death in pancreatitis. Manipulating these signals to change the pattern of death responses presents a therapeutic strategy for treatment of pancreatitis. PMID- 16339140 TI - Prediction of mitochondrial proteins using support vector machine and hidden Markov model. AB - Mitochondria are considered as one of the core organelles of eukaryotic cells hence prediction of mitochondrial proteins is one of the major challenges in the field of genome annotation. This study describes a method, MitPred, developed for predicting mitochondrial proteins with high accuracy. The data set used in this study was obtained from Guda, C., Fahy, E. & Subramaniam, S. (2004) Bioinformatics 20, 1785-1794. First support vector machine-based modules/methods were developed using amino acid and dipeptide composition of proteins and achieved accuracy of 78.37 and 79.38%, respectively. The accuracy of prediction further improved to 83.74% when split amino acid composition (25 N-terminal, 25 C terminal, and remaining residues) of proteins was used. Then BLAST search and support vector machine-based method were combined to get 88.22% accuracy. Finally we developed a hybrid approach that combined hidden Markov model profiles of domains (exclusively found in mitochondrial proteins) and the support vector machine-based method. We were able to predict mitochondrial protein with 100% specificity at a 56.36% sensitivity rate and with 80.50% specificity at 98.95% sensitivity. The method estimated 9.01, 6.35, 4.84, 3.95, and 4.25% of proteins as mitochondrial in Saccharomyces cerevisiae, Drosophila melanogaster, Caenorhabditis elegans, mouse, and human proteomes, respectively. MitPred was developed on the above hybrid approach. PMID- 16339141 TI - COX24 codes for a mitochondrial protein required for processing of the COX1 transcript. AB - In most strains of Saccharomyces cerevisiae the mitochondrial gene COX1, for subunit 1 of cytochrome oxidase, contains multiple exons and introns. Processing of COX1 primary transcript requires accessory proteins factors, some of which are encoded by nuclear genes and others by reading frames residing in some of the introns of the COX1 and COB genes. Here we show that the low molecular weight protein product of open reading frame YLR204W, for which we propose the name COX24, is also involved in processing of COX1 RNA intermediates. The growth defect of cox24 mutants is partially rescued in strains harboring mitochondrial DNA lacking introns. Northern blot analyses of mitochondrial transcripts indicate cox24 null mutants to be blocked in processing of introns aI2 and aI3. The dependence of intron aI3 excision on Cox24p is also supported by the growth properties of the cox24 mutant harboring mitochondrial DNA with different intron compositions. The intermediate phenotype of the cox24 mutant in the background of intronless mitochondrial DNA, however, suggests that in addition to its role in splicing of the COX1 pre-mRNA, Cox24p still has another function. Based on the analysis of a cox14-cox24 double mutant, we propose that the other function of Cox24p is related to translation of the COX1 mRNA. PMID- 16339142 TI - Rac1 modulates sphingosine 1-phosphate-mediated activation of phosphoinositide 3 kinase/Akt signaling pathways in vascular endothelial cells. AB - Sphingosine 1-phosphate (S1P) is a platelet-derived sphingolipid that activates G protein-coupled S1P receptors and initiates a broad range of responses in vascular endothelial cells. The small GTPase Rac1 is implicated in diverse S1P modulated cellular responses in endothelial cells, yet the molecular mechanisms involved in S1P-mediated Rac1 activation are incompletely understood. We studied the pathways involved in S1P-mediated Rac1 activation in bovine aortic endothelial cells (BAEC) and found that S1P-induced Rac1 activation is impaired following chelation of G protein betagamma subunits by transfection of betaARKct. Treatment with the Src tyrosine kinase inhibitor PP2 completely attenuated S1P mediated Rac1 activation; however, pretreatment of BAEC with wortmannin, an inhibitor of phosphoinositide (PI) 3-kinase, had no effect on Rac1 activation while completely blocking S1P-induced Akt phosphorylation. We used Rac1-specific small interfering RNA (siRNA) duplexes to "knock down" endogenous Rac1 expression and found that siRNA-mediated Rac1 knockdown significantly impaired basal as well as S1P-induced phosphorylation of protein kinase Akt, as well as several downstream targets of Akt including endothelial nitric-oxide synthase and glycogen synthase kinase 3beta. By contrast, S1P-induced phosphorylation of the mitogen-activated protein kinases ERK1/2 was unperturbed by siRNA-mediated Rac1 knockdown. We found that overexpression of the Rac1 guanine nucleotide exchange factor (GEF) Tiam1 markedly enhanced Rac1 activity, whereas a dominant negative Tiam1 mutant significantly attenuated S1P-mediated Rac1 activation. Taken together, these studies identify G protein betagamma subunits, Src kinase and the GEF Tiam1 as upstream modulators of S1P-mediated Rac1 activation, and establish a central role for Rac1 in S1P-mediated activation of PI 3-kinase/Akt/endothelial nitric-oxide synthase signaling in vascular endothelial cells. PMID- 16339143 TI - Diverse effects of pathogenic mutations of Parkin that catalyze multiple monoubiquitylation in vitro. AB - Mutational dysfunction of PARKIN gene, which encodes a double RING finger protein and has ubiquitin ligase E3 activity, is the major cause of autosomal recessive juvenile Parkinsonism. Although many studies explored the functions of Parkin, its biochemical character is poorly understood. To address this issue, we established an E3 assay system using maltose-binding protein-fused Parkin purified from Escherichia coli. Using this recombinant Parkin, we found that not the front but the rear RING finger motif is responsible for the E3 activity of Parkin, and it catalyzes multiple monoubiquitylation. Intriguingly, for autosomal recessive juvenile Parkinsonism-causing mutations of Parkin, whereas there was loss of E3 activity in the rear RING domain, other pathogenic mutants still exhibited E3 activity equivalent to that of the wild-type Parkin. The evidence presented allows us to reconsider the function of Parkin-catalyzed ubiquitylation and to conclude that autosomal recessive juvenile Parkinsonism is not solely attributable to catalytic impairment of the E3 activity of Parkin. PMID- 16339144 TI - Differential regulation of cardiomyocyte survival and hypertrophy by MDM2, an E3 ubiquitin ligase. AB - MDM2 is an E3 ubiquitin ligase that regulates the proteasomal degradation and activity of proteins involved in cell growth and apoptosis, including the tumor suppressors p53 and retinoblastoma and the transcription factor E2F1. Although the effect of several MDM2 targets on cardiomyocyte survival and hypertrophy has already been investigated, the role of MDM2 in these processes has not yet been established. We have, therefore, analyzed the effect of overexpression as well as inhibition of MDM2 on cardiac ischemia/reperfusion injury and hypertrophy. Here we show that isolated cardiac myocytes overexpressing MDM2 acquired resistance to hypoxia/reoxygenation-induced cell death. Conversely, inactivation of MDM2 by a peptide inhibitor resulted in elevated p53 levels and promoted hypoxia/reoxygenation-induced apoptosis. Consistent with this, decreased expression of MDM2 in a genetic mouse model was accompanied by reduced functional recovery of the left ventricles determined with the Langendorff ex vivo model of ischemia/reperfusion. In contrast to cell survival, cell hypertrophy induced by the alpha-agonists phenylephrine or endothelin-1 was inhibited by MDM2 overexpression. Collectively, our studies indicate that MDM2 promotes survival and attenuates hypertrophy of cardiac myocytes. This differential regulation of cell growth and cell survival is unique, because most other survival factors are prohypertrophic. MDM2, therefore, might be a potential therapeutic target to down regulate both cell death and pathologic hypertrophy during remodeling upon cardiac infarction. In addition, our data also suggest that cancer treatments with MDM2 inhibitors to reactivate p53 may have adverse cardiac side effects by promoting cardiomyocyte death. PMID- 16339145 TI - Vitamin B6 biosynthesis by the malaria parasite Plasmodium falciparum: biochemical and structural insights. AB - Vitamin B6 is one of nature's most versatile cofactors. Most organisms synthesize vitamin B6 via a recently discovered pathway employing the proteins Pdx1 and Pdx2. Here we present an in-depth characterization of the respective orthologs from the malaria parasite, Plasmodium falciparum. Expression profiling of Pdx1 and -2 shows that blood-stage parasites indeed possess a functional vitamin B6 de novo biosynthesis. Recombinant Pdx1 and Pdx2 form a complex that functions as a glutamine amidotransferase with Pdx2 as the glutaminase and Pdx1 as pyridoxal-5 ' phosphate synthase domain. Complex formation is required for catalytic activity of either domain. Pdx1 forms a chimeric bi-enzyme with the bacterial YaaE, a Pdx2 ortholog, both in vivo and in vitro, although this chimera does not attain full catalytic activity, emphasizing that species-specific structural features govern the interaction between the protein partners of the PLP synthase complexes in different organisms. To gain insight into the activation mechanism of the parasite bi-enzyme complex, the three-dimensional structure of Pdx2 was determined at 1.62 A. The obstruction of the oxyanion hole indicates that Pdx2 is in a resting state and that activation occurs upon Pdx1-Pdx2 complex formation. PMID- 16339146 TI - SARS coronavirus, but not human coronavirus NL63, utilizes cathepsin L to infect ACE2-expressing cells. AB - Viruses require specific cellular receptors to infect their target cells. Angiotensin-converting enzyme 2 (ACE2) is a cellular receptor for two divergent coronaviruses, SARS coronavirus (SARS-CoV) and human coronavirus NL63 (HCoV NL63). In addition to hostcell receptors, lysosomal cysteine proteases are required for productive infection by some viruses. Here we show that SARS-CoV, but not HCoV-NL63, utilizes the enzymatic activity of the cysteine protease cathepsin L to infect ACE2-expressing cells. Inhibitors of cathepsin L blocked infection by SARS-CoV and by a retrovirus pseudotyped with the SARS-CoV spike (S) protein but not infection by HCoV-NL63 or a retrovirus pseudotyped with the HCoV NL63 S protein. Expression of exogenous cathepsin L substantially enhanced infection mediated by the SARS-CoV S protein and by filovirus GP proteins but not by the HCoV-NL63 S protein or the vesicular stomatitis virus G protein. Finally, an inhibitor of endosomal acidification had substantially less effect on infection mediated by the HCoV-NL63 S protein than on that mediated by the SARS CoV S protein. Our data indicate that two coronaviruses that utilize a common receptor nonetheless enter cells through distinct mechanisms. PMID- 16339147 TI - Mutations in the amino terminus of the cystic fibrosis transmembrane conductance regulator enhance endocytosis. AB - Efficient endocytosis of the cystic fibrosis transmembrane conductance regulator (CFTR) is mediated by a tyrosine-based internalization signal in the CFTR carboxyl-terminal tail 1424YDSI1427. In the present studies, two naturally occurring cystic fibrosis mutations in the amino terminus of CFTR, R31C, and R31L were examined. To determine the defect that these mutations cause, the Arg-31 mutants were expressed in COS-7 cells and their biogenesis and trafficking to the cell surface tested in metabolic pulse-chase and surface biotinylation assays, respectively. The results indicated that both Arg-31 mutants were processed to band C at approximately 50% the efficiency of the wild-type protein. However, once processed and delivered to the cell surface, their half-lives were the same as wild-type protein. Interestingly, indirect immunofluorescence and cell surface biotinylation indicated that the surface pool was much smaller than could be accounted for based on the biogenesis defect alone. Therefore, the Arg-31 mutants were tested in internalization assays and found to be internalized at 2x the rate of the wild-type protein. Patch clamp and 6-methoxy-N-(3-sulfopropyl)quinolinium analysis confirmed reduced amounts of functional Arg-31 channels at the cell surface. Together, the results suggest that both R31C and R31L mutations compromise biogenesis and enhance internalization of CFTR. These two additive effects contribute to the loss of surface expression and the associated defect in chloride conductance that is consistent with a disease phenotype. PMID- 16339148 TI - Pigment epithelium-derived factor inhibits angiogenesis via regulated intracellular proteolysis of vascular endothelial growth factor receptor 1. AB - Pigment epithelium-derived factor (PEDF) has been identified as one of the most potent of endogenous negative regulators of blood vessel growth in the body. Here we report that PEDF is able to inhibit growth factor-induced angiogenesis in microvascular endothelial cells through a novel pathway requiring cleavage and intracellular translocation of the transmembrane domain of the VEGFR-1. Analysis of the subcellular distribution of VEGFR-1 revealed the appearance of an 80-kDa C terminal domain in the cytosol of cells treated with VEGF and PEDF that correlated with a decrease of the full-length receptor in the nuclear and cytoskeletal fractions. This regulated intramembrane proteolysis is dependent on gamma-secretase because inhibition of gamma-secretase abolished the inhibitory effect of PEDF on VEGF-induced angiogenesis as well as VEGFR-1 cleavage. The addition of PEDF to microvascular endothelial cells significantly increases gamma secretase activity even in the absence of VEGF, showing that VEGF binding to VEGF R1 is essential for substrate availability. This increase in activity was associated with translocation of presenilin 1 from the perinuclear region to the cell membrane. PEDF was also able to inhibit VEGF-induced phosphorylation of VEGFR-1. Taken together we have identified two novel pathways by which PEDF inhibits VEGF-induced angiogenesis: regulated intramembrane proteolysis and inhibition of phosphorylation. This confirms the importance of PEDF and VEGFR-1 in the negative regulation of angiogenesis. PMID- 16339149 TI - Identification of fungal sphingolipid C9-methyltransferases by phylogenetic profiling. AB - Fungal glucosylceramides play an important role in plant-pathogen interactions enabling plants to recognize the fungal attack and initiate specific defense responses. A prime structural feature distinguishing fungal glucosylceramides from those of plants and animals is a methyl group at the C9-position of the sphingoid base, the biosynthesis of which has never been investigated. Using information on the presence or absence of C9-methylated glucosylceramides in different fungal species, we developed a bioinformatics strategy to identify the gene responsible for the biosynthesis of this C9-methyl group. This phylogenetic profiling allowed the selection of a single candidate out of 24-71 methyltransferase sequences present in each of the fungal species with C9 methylated glucosylceramides. A Pichia pastoris knock-out strain lacking the candidate sphingolipid C9-methyltransferase was generated, and indeed, this strain contained only non-methylated glucosylceramides. In a complementary approach, a Saccharomyces cerevisiae strain was engineered to produce glucosylceramides suitable as a substrate for C9-methylation. C9-methylated sphingolipids were detected in this strain expressing the candidate from P. pastoris, demonstrating its function as a sphingolipid C9-methyltransferase. The enzyme belongs to the superfamily of S-adenosylmethionine-(SAM)-dependent methyltransferases and shows highest sequence similarity to plant and bacterial cyclopropane fatty acid synthases. An in vitro assay showed that sphingolipid C9 methylation is membrane-bound and requires SAM and Delta4,8-desaturated ceramide as substrates. PMID- 16339150 TI - The Drosophila fused lobes gene encodes an N-acetylglucosaminidase involved in N glycan processing. AB - Most processed, e.g. fucosylated, N-glycans on insect glycoproteins terminate in mannose, yet the relevant modifying enzymes require the prior action of N acetylglucosaminyltransferase I. This led to the hypothesis that a hexosaminidase acts during the course of N-glycan maturation. To determine whether the Drosophila melanogaster genome indeed encodes such an enzyme, a cDNA corresponding to fused lobes (fdl), a putative beta-N-acetylglucosaminidase with a potential transmembrane domain, was cloned. When expressed in Pichia pastoris, the enzyme exhibited a substrate specificity similar to that previously described for a hexosaminidase activity from Sf-9 cells, i.e. it hydrolyzed exclusively the GlcNAc residue attached to the alpha1,3-linked mannose of the core pentasaccharide of N-glycans. It also hydrolyzed p-nitrophenyl-N-acetyl-beta glucosaminide, but not chitooligosaccharides; in contrast, Drosophila HEXO1 and HEXO2 expressed in Pichia cleaved both these substrates but not N-glycans. The localization of recombinant FDL tagged with green fluorescent protein in Drosophila S2 cells by immunoelectron microscopy showed that this enzyme transits through the Golgi, is present on the plasma membrane and in multivesicular bodies, and is secreted. Finally, the N-glycans of two lines of fdl mutant flies were analyzed by mass spectrometry and reversed-phase high-performance liquid chromatography. The ratio of structures with terminal GlcNAc over those without (i.e. paucimannosidic N-glycans) was drastically increased in the fdl-deficient flies. Therefore, we conclude that the fdl gene encodes a novel hexosaminidase responsible for the occurrence of paucimannosidic N-glycans in Drosophila. PMID- 16339151 TI - The human anti-apoptotic proteins cIAP1 and cIAP2 bind but do not inhibit caspases. AB - cIAPs (cellular inhibitor of apoptosis proteins) 1 and 2 are able to regulate apoptosis when ectopically expressed in recipient cells and probably also in vivo. Previous work suggested that this is at least partially due to direct caspase inhibition, mediated by two of the three baculovirus IAP repeat (BIR) domains that are contained in these proteins. In support of this we show that the BIR domains 2 and 3 of the two cIAPs are able to bind caspases-7 and -9. However, we demonstrate that neither of these BIR domains is able to inhibit caspases because of critical substitutions in the regions that target caspase inhibition in the X-linked IAP, a tight binding caspase inhibitor. The cIAP BIR domains can be converted to tight binding caspase inhibitors by substituting these critical residues with XIAP residues. Thus, cIAPs maintain protein scaffolds suitable for direct caspase inhibition but have lost or never acquired specific caspase inhibitory interaction sites. Consequently, although the binding function of the cIAP BIRs may be important for their physiologic function, caspase inhibition is not. PMID- 16339152 TI - Characterization of the bifunctional gamma-glutamate-cysteine ligase/glutathione synthetase (GshF) of Pasteurella multocida. AB - Glutamate-cysteine ligase (gamma-ECL) and glutathione synthetase (GS) are the two unrelated ligases that constitute the glutathione biosynthesis pathway in most eukaryotes, purple bacteria, and cyanobacteria. gamma-ECL is a member of the glutamine synthetase family, whereas GS enzymes group together with highly diverse carboxyl-to-amine/thiol ligases, all characterized by the so-called two domain ATP-grasp fold. This generalized scheme toward the formation of glutathione, however, is incomplete, as functional steady-state levels of intracellular glutathione may also accumulate solely by import, as has been reported for the Pasteurellaceae member Haemophilus influenzae, as well as for certain Gram-positive enterococci and streptococci, or by the action of a bifunctional fusion protein (termed GshF), as has been reported recently for the Gram-positive firmicutes Streptococcus agalactiae and Listeria monocytogenes. Here, we show that yet another member of the Pasteurellaceae family, Pasteurella multocida, acquires glutathione both by import and GshF-driven biosynthesis. Domain architecture analysis shows that this P. multocida GshF bifunctional ligase contains an N-terminal gamma-proteobacterial gamma-ECL-like domain followed by a typical ATP-grasp domain, which most closely resembles that of cyanophycin synthetases, although it has no significant homology with known GS ligases. Recombinant P. multocida GshF overexpresses as an approximately 85-kDa protein, which, on the basis of gel-sizing chromatography, forms dimers in solution. The gamma-ECL activity of GshF is regulated by an allosteric type of glutathione feedback inhibition (K(i) = 13.6 mM). Furthermore, steady-state kinetics, on the basis of which we present a novel variant of half-of-the-sites reactivity, indicate intimate domain-domain interactions, which may explain the bifunctionality of GshF proteins. PMID- 16339154 TI - Determination of the substrate specificity of tripeptidyl-peptidase I using combinatorial peptide libraries and development of improved fluorogenic substrates. AB - Classical late-infantile neuronal ceroid lipofuscinosis is a fatal neurodegenerative disease caused by mutations in CLN2, the gene encoding the lysosomal protease tripeptidyl-peptidase I (TPP I). The natural substrates for TPP I and the pathophysiological processes associated with lysosomal storage and disease progression are not well understood. Detailed characterization of TPP I substrate specificity should provide insights into these issues and also aid in the development of improved clinical and biochemical assays. To this end, we constructed fluorogenic and standard combinatorial peptide libraries and analyzed them using fluorescence and mass spectrometry-based activity assays. The fluorogenic group 7-amino-4-carbamoylmethylcoumarin was incorporated into a series of 7-amino-4-carbamoylmethylcoumarin tripeptide libraries using a design strategy that allowed systematic evaluation of the P1, P2, and P3 positions. TPP I digestion of these substrates liberates the fluorescence group and results in a large increase in fluorescence that can be used to calculate kinetic parameters and to derive the substrate specificity constant kcat/KM. In addition, we implemented a mass spectrometry-based assay to measure the hydrolysis of individual peptides in peptide pools and thus expand the scope of the analysis. Nonfluorogenic tetrapeptide and pentapeptide libraries were synthesized and analyzed to evaluate P1' and P2' residues. Together, this analysis allowed us to predict the relative specificity of TPP I toward a wide range of potential biological substrates. In addition, we evaluated a variety of new fluorogenic peptides with a P3 Arg residue, and we demonstrated their superiority compared with the widely used substrate Ala-Ala-Phe-AMC for selectively measuring TPP I activity in biological specimens. PMID- 16339155 TI - A major cell wall lipopeptide of Mycobacterium avium subspecies paratuberculosis. AB - Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne disease in cattle and other ruminants, is proposed to be at least one of the causes of Crohn disease in humans. MAP and Mycobacterium avium subspecies avium, a closely related opportunistic environmental bacterium, share 95% of their genes and exhibit homologies of more than 99% between these genes. The identification of molecules specific for MAP is essential for understanding its pathogenicity and for development of useful diagnostic tools. The application of gas chromatography, mass spectrometry, and nuclear magnetic resonance led to the structural identification of a major cell wall lipopeptide of MAP, termed Para-LP 01, defined as C20 fatty acyl-D-Phe-N-Me-L-Val-L-Ile-L-Phe-L-Ala methyl ester. Variations of this lipopeptide with different fatty acyl moieties (C16 fatty acyl through C17, C18, C19, C21 to C22) were also identified. Besides the specificity of this lipopeptide for MAP, the presence of an N-Me-L-valine represents the first reported N-methylated amino acid within an immunogenic lipopeptide of mycobacteria. Sera from animals with Johne disease, but not sera from uninfected cattle, reacted with this lipopeptide, indicating potential biological importance. PMID- 16339153 TI - Kinetic analysis of a mammalian phospholipase D: allosteric modulation by monomeric GTPases, protein kinase C, and polyphosphoinositides. AB - In mammalian cells, phospholipase D activity is tightly regulated by diverse cellular signals, including hormones, neurotransmitters, and growth factors. Multiple signaling pathways converge upon phospholipase D to modulate cellular actions, such as cell growth, shape, and secretion. We examined the kinetics of protein kinase C and G-protein regulation of mammalian phospholipase D1 (PLD1) in order to better understand interactions between PLD1 and its regulators. Activation by Arf-1, RhoA, Rac1, Cdc42, protein kinase Calpha, and phosphatidylinositol 4,5-bisphosphate displayed surface dilution kinetics, but these effectors modulated different kinetic parameters. PKCalpha activation of PLD1 involves N- and C-terminal PLD domains. Rho GTPases were binding activators, enhancing the catalytic efficiency of a purified PLD1 catalytic domain via effects on Km. Arf-1, a catalytic activator, stimulated PLD1 by enhancing the catalytic constant, kcat. A kinetic description of PLD1 activation by multiple modulators reveals a mechanism for apparent synergy between activators. Synergy was observed only when PLD1 was simultaneously stimulated by a binding activator and a catalytic activator. Surprisingly, synergistic activation was steeply dependent on phosphatidylinositol 4,5-bisphosphate and phosphatidylcholine. Together, these findings suggest a role for PLD1 as a signaling node, in which integration of convergent signals occurs within discrete locales of the cellular membrane. PMID- 16339156 TI - Characterization of distinct Stat5b binding sites that mediate growth hormone stimulated IGF-I gene transcription. AB - A key agent in the anabolic actions of growth hormone (GH) is insulin-like growth factor-I (IGF-I), a 70-amino acid secreted protein with direct effects on somatic growth and tissue maintenance and repair. GH rapidly and potently stimulates IGF I gene transcription by mechanisms independent of new protein synthesis, and recent studies have linked the transcription factor Stat5b to a regulatory network connecting the activated GH receptor on the cell membrane to the IGF-I gene in the nucleus. Here we analyze two distinct conserved GH response elements in the rat IGF-I locus that contain paired Stat5b sites. Each response element binds Stat5b in vivo in a GH-dependent way, as assessed by chromatin immunoprecipitation assays, and consists of one high affinity and one lower affinity Stat5b site, as determined by both qualitative and quantitative protein DNA binding studies. In biochemical reconstitution experiments, both response elements are able to mediate GH-stimulated and Stat5b-dependent transcription when fused to a reporter gene containing either the major IGF-I promoter or a minimal neutral promoter, although the paired Stat5b sites located in the second IGF-I intron were more than twice as effective as the response element that mapped approximately 73 kb 5' to the IGF-I exon 1. Taken together, our results define the initial molecular architecture of a complicated GH-regulated transcriptional pathway, and suggest that apparently redundant hormone response elements provide a mechanism for amplifying GH action at a physiologically important target gene. PMID- 16339158 TI - Rehabilitating digoxin. PMID- 16339157 TI - Digoxin and reduction in mortality and hospitalization in heart failure: a comprehensive post hoc analysis of the DIG trial. AB - AIMS: To determine the effects of digoxin on all-cause mortality and heart failure (HF) hospitalizations, regardless of ejection fraction, accounting for serum digoxin concentration (SDC). METHODS AND RESULTS: This comprehensive post hoc analysis of the randomized controlled Digitalis Investigation Group trial (n=7788) focuses on 5548 patients: 1687 with SDC, drawn randomly at 1 month, and 3861 placebo patients, alive at 1 month. Overall, 33% died and 31% had HF hospitalizations during a 40-month median follow-up. Compared with placebo, SDC 0.5-0.9 ng/mL was associated with lower mortality [29 vs. 33% placebo; adjusted hazard ratio (AHR), 0.77; 95% confidence interval (CI), 0.67-0.89], all-cause hospitalizations (64 vs. 67% placebo; AHR, 0.85; 95% CI, 0.78-0.92) and HF hospitalizations (23 vs. 33% placebo; AHR, 0.62; 95% CI, 0.54-0.72). SDC> or =1.0 ng/mL was associated with lower HF hospitalizations (29 vs. 33% placebo; AHR, 0.68; 95% CI, 0.59-0.79), without any effect on mortality. SDC 0.5-0.9 reduced mortality in a wide spectrum of HF patients and had no interaction with ejection fraction >45% (P=0.834) or sex (P=0.917). CONCLUSIONS: Digoxin at SDC 0.5-0.9 ng/mL reduces mortality and hospitalizations in all HF patients, including those with preserved systolic function. At higher SDC, digoxin reduces HF hospitalization but has no effect on mortality or all-cause hospitalizations. PMID- 16339159 TI - Protecting Europeans from secondhand smoke: time to act. PMID- 16339160 TI - Role of HCV coinfection towards disease progression and survival in HIV-1 infected children: a follow-up study of 10 years. AB - The present study was taken up to evaluate the pattern of disease progression and survival in a group of HIV-1 positive children, coinfected with HCV infection (n=25) in comparison to those without such coinfection (n=23). There was a significant negative correlation between the rate of decline of the CD4 + T cell percentage and the duration of the AIDS-free interval in most (80.0 per cent) of the HCV seropositive children showing such decline (r=-0.588; p=0.005). The HCV seropositive children had twofold higher risk of progression to development of AIDS than HCV seronegatives (RR=2.51; 95 per cent CI:1.34-4.69; p=0.004). There was a significant negative correlation between the rate of decline of CD4 + T cell percentage and overall survival duration for HCV seropositive group (r= 0.609; p=0.003). Moreover, children coinfected with HCV had more than twofold higher risks of death than those without HCV (RR=2.39; 95 per cent CI:1.17-4.89; p<0.01). It appears that HCV infection may be an important contributor to the rapid disease progression and increase in mortality in HCV-HIV-1 coinfected children of thalassemia major. PMID- 16339161 TI - The ERA-EDTA cohort study--comparison of methods to predict survival on renal replacement therapy. AB - BACKGROUND: Accurate prediction of patient survival from the time of starting renal replacement therapy (RRT) is desirable, but previously published predictive models have low accuracy. We have attempted to overcome limitations of previous studies by conducting an ambidirectional inception cohort study in patients on RRT from centres throughout Europe. A conventional multivariate regression (MVR) model, a self-learning rule-based model (RBM) and a simple co-morbidity score [the Charlson score modified for renal disease (MCS)] were compared. METHODS: In 1996, all 3640 dialysis centres registered with the ERA-EDTA were invited to identify all patients on RRT for end-stage renal failure (ESRF) who died during the 28 days of February 1997 (training cohort) and all patients who started RRT in the same period (validation cohort). Fifty-four clinical and laboratory variables from the time of starting RRT were collected in both cohorts using a two-page questionnaire. The data from the training cohort were given to statisticians at the Amsterdam Academic Medical Centre to create the MVR model and to engineers in Strathclyde University to create the RBM. They were then given the baseline data from patients in the validation cohort to predict how long each patient would survive. Follow-up questionnaires were sent to the centre of each patient in the validation cohort to determine actual survival. RESULTS: A total of 2310 patients from 793 centres in 37 countries in the ERA-EDTA area were used to construct and validate the models. For predicting 1-year survival, the RBM had the highest positive predictive value (PPV) (84.2%), the MVR model had the highest negative predictive value (NPV) (47%) and the RBM had the highest likelihood ratio (1.59). For predicting 5-year survival, the MCS had the highest PPV (79.4%), the RBM had the highest NPV (74.3%) and the MCS had the highest likelihood ratio (7.0). The proportion of explained variance in survival for MCS, MVR and RBM was 14.6, 12.9 and 3.95%, respectively. CONCLUSION: Using the ambidirectional inception cohort design of this ERA-EDTA Registry survey, we have been able to create and validate two novel instruments to predict survival in patients starting RRT and compare them with a simple scoring model. The models tended to predict 5-year survival with more accuracy than 1-year survival. Examples of potential applications include informing clinical decision making about the likely benefit of starting RRT and listing for transplantation, adjusting for baseline risk in comparative studies and identifying specific risk groups to participate in clinical trials. PMID- 16339162 TI - Pregnancy of a lupus patient--a challenge to the nephrologist. PMID- 16339163 TI - Leptospiral membrane proteins stimulate pro-inflammatory chemokines secretion by renal tubule epithelial cells through toll-like receptor 2 and p38 mitogen activated protein kinase. AB - BACKGROUND: Leptospiral membrane proteins extracted from pathogenic Leptospira santarosai serovar Shermani (LMPS) stimulated pro-inflammatory chemokines production in cultured mouse proximal tubule epithelial cells (PTECs) and implicated its role in the pathogenesis of leptospira-induced tubulointerstitial nephritis. PTECs express the functional TLR2 and TLR4, which have been shown to play essential roles in innate immunity. This study investigated the roles of Toll-like receptors (TLRs) and mitogen-activated protein kinases (MAPKs) signalling pathways in the pathogenesis of leptospira-induced tubulointerstitial nephritis. METHODS: The immortalized mouse PKSV-PR late PTECs were used as the model system. The genes expression and secretion of CCL2/monocyte chemoattractant protein-1 (CCL2/MCP-1) and CXCL2/macrophage inflammatory protein-2 (CXCL2/MIP-2) were measured by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). We investigated MAPKs signalling pathways by Western blot and their reciprocal roles by specific inhibitors. A specific TLR2 neutralizing antibody was applied to evaluate the crosstalk between TLR2 and MAPKs. RESULTS: The LMPS stimulated extracellular signal-regulated kinases (ERK1/2), c-Jun N-terminal kinases (JNKs) and p38 mitogen-activated protein kinase (p38 MAPK), initiated the nuclear transcription factor kappaB (NF kappaB), and enhanced the secretion of CCL2/MCP-1 and CXCL2/MIP-2. The LMPS also unregulated the level of TLR2 mRNA expression in PTECs through time- and dose dependent effects. The LMPS enhanced the secretion of CCL2/MCP-1 and CXCL8/interleukin-8 (CXCL8/IL-8) in TLR-defective human embryonic kidney (HEK) 293 cells only when transfected with a TLR2 expressing plasmid. The secretions of CCL2/MCP-1 and CXCL2/MIP-2 stimulated by LMPS were significantly reduced by incubating PTECs with SB203580, an inhibitor of p38 MAPK. Furthermore, a neutralizing anti-mouse TLR2 antibody hindered the phosphorylation of p38 and LMPS-stimulated secretion of CCL2/MCP-1 and CXCL2/MIP-2. CONCLUSION: These findings demonstrate that activation of p38 MAPK and release of chemokines by LMPS are mediated by TLR2 in renal proximal tubule cells. These results also implicate the crucial role of innate immunity in leptospira-induced tubulointerstitial nephritis. PMID- 16339164 TI - Does a late referral to a nephrologist constitute a problem in children starting renal replacement therapy in Poland?--a nationwide study. AB - BACKGROUND: It is estimated that 20-50% of adult patients start chronic dialysis therapy without prior contact with a nephrologist. The aim of this nationwide study was to assess clinical and metabolic status of children at the start of chronic dialysis in Poland with regard to the timing of the referral to a nephrologist. METHODS: We studied data of 180 children (mean age 14+/-6 years) undergoing chronic dialysis in 13 (out of 14) dialysis pediatric centres in Poland. Patients were classified as early referrals (ERs) when they entered the dialysis programme at least 1 month after the first referral to a nephrologist or late referrals (LRs) when the dialysis was introduced within 1 month from the first visit. RESULTS: Seventy-nine percent of pediatric patients were referred early (ER) to the dialysis centre and 21% were referred late (LR) and had to start dialysis within a month. When starting dialysis, LR patients had significantly higher levels of urea and phosphate as well as lower calcium and haemoglobin in comparison with ERs. Hypertension, pulmonary oedema, fluid overload, treatment in the intensive care unit (ICU) and body mass index (BMI) below 10th percentile turned out to be more frequent in the LR group. Peritoneal dialysis (PD) was used as the first method of dialysis in 59% of ERs and 46% of LRs. The majority of ER patients was treated in the predialysis period with calcitriol, phosphate binders and low protein diet (84%, 89%, 92% of all children, respectively), and 20% of them received epoetin. In the up to 3 years observation of our initial cohort, we also found that the patients who were referred late were less likely to receive kidney transplant (P = 0.02). CONCLUSION: The results of the study indicate that the LR to a pediatric nephrologist was associated with poorer clinical and metabolic status of children entering chronic dialysis programmes. PMID- 16339165 TI - Perinatal outcome of 12,021 singleton and 3108 twin births after non-IVF-assisted reproduction: a cohort study. AB - Perinatal outcome of pregnancies caused by assisted reproduction technique (ART) is substantially worse when compared with pregnancies following natural conception. We investigated the possible risks of non-IVF ART on perinatal health. We conducted a retrospective cohort study with two exposure groups: a study group of pregnancies after controlled ovarian stimulation (COS), with or without artificial insemination (AI), and a naturally conceived comparison group. We used the data from the regional registry of all hospital deliveries in the Dutch-speaking part of Belgium during the period from January 1993 until December 2003 to investigate differences in perinatal outcome of singleton and twin pregnancies. 12 021 singleton and 3108 twin births could be selected. Naturally conceived subjects were matched for maternal age, parity, fetal sex and year of birth. The main outcome measures were duration of pregnancy, birth weight, perinatal morbidity and perinatal mortality. Our overall results showed a significantly higher incidence of prematurity (<32 and <37 weeks), low and very low birth weight, transfer to the neonatal intensive care unit and most neonatal morbidity parameters for COS/AI singletons. Twin pregnancies resulting from COS/AI showed an increased rate of neonatal mortality, assisted ventilation and respiratory distress syndrome. After excluding same-sex twin sets, COS/AI twin pregnancies were at increased risk for extreme prematurity and very low birth weight. In conclusion, COS/AI singleton and twin pregnancies are significantly disadvantaged compared to naturally conceived children. PMID- 16339166 TI - Menstruation-free interval and ongoing pregnancy in IVF using GnRH antagonists. AB - BACKGROUND: The purpose of this study was to evaluate prospectively the association between the achievement of ongoing pregnancy and the time interval from the end of menstruation until the administration of HCG (menstruation-free interval) in patients treated by IVF. METHODS: A fixed dose of 200 IU of recombinant FSH (rFSH) was started in 90 patients on day 2 of the menstrual cycle and daily GnRH antagonist was initiated on day 6 of stimulation. Triggering of final oocyte maturation was performed with 10,000 IU of HCG as soon as three follicles of > or =17 mm were present at ultrasound. RESULTS: Single embryo transfer was performed in 64.6% of the patients who reached embryo transfer (53/82). Ongoing pregnancy rate per embryo transfer was 18.3% (95% CI 11.4 28.0%). The menstruation-free interval significantly predicted the probability of ongoing pregnancy in a logistic regression analysis, controlling for female age and LH on day 1 of stimulation (odds ratio for the menstruation-free interval: 0.70; 95% CI: 0.54-0.92). CONCLUSION: The longer the interval from the end of menstruation until the administration of HCG, the lower the probability of ongoing pregnancy in patients stimulated with recombinant FSH and GnRH antagonist for IVF. PMID- 16339167 TI - A new Dutch Law regulating provision of identifying information of donors to offspring: background, content and impact. AB - In 2004 a law was introduced in The Netherlands, which gives offspring conceived by semen or oocyte donation the right to know the identity of the donor. The law also regulates the provision of other information concerning the donor to the offspring, their parents or their general practitioner. With the introduction of this law, a choice has been made in which the wish of offspring prevails above others involved. Donors can no longer claim absolute anonymity; they are anonymous at the time of donation, but if a child aged > or =16 years requests information the donor may now be traced. During 15 years of debate on the abolition of donor anonymity the number of donors decreased by >70% and the number of semen banks by 50%. We describe the debate which led to the law, the characteristics of the law itself and note some of the probable and possible consequences for donor offspring, parents, donors and semen banks. PMID- 16339168 TI - Study of two markers of apoptosis and meiotic segregation in ejaculated sperm of chromosomal translocation carrier patients. AB - BACKGROUND: To try to explain the infertility of chromosomal translocation carrier patients, we compared the expression of two markers of apoptosis in the sperm of patients and of fertile donors, and we studied the meiotic segregation in the ejaculated sperm of these translocation carriers. METHODS: Twenty semen samples of translocation carriers, [reciprocal (n=14) and Robertsonian translocations (n=6)], were compared with the semen samples of donors (n=20). Different tests were applied: annexin V binding assay; terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL); and fluorescence in situ hybridization (FISH). RESULTS: The annexin V binding assay in sperm of patients with chromosomal translocation (n=17) showed a significantly increased proportion of sperm with externalized phosphatidylserine (PS) than in the control group (n=20, P2 months) female mice were ovariectomized and treated for 8-12 d with implants containing estradiol (E), E and P, E and DHT, or E, P, and DHT. Targeted extracellular recordings were used to examine the long-term firing activity of green fluorescent protein-identified GnRH neurons in brain slices from these mice. In comparing E alone to E plus P animals, P increased the percentage of time that GnRH neurons were quiescent and reduced the area under the curve of the firing rate and the instantaneous firing frequency, suggesting that P provides additional negative feedback over E alone. The addition of DHT markedly increased GnRH neuron activity in both the presence and absence of P. DHT also altered the firing pattern of GnRH neurons, such that peaks in the firing rate detected by the Cluster8 algorithm were approximately doubled in frequency and amplitude. These data support and extend our previous findings and are consistent with the hypothesis that the changes in GABAergic transmission observed in these animal models impact upon the activity of GnRH neurons, and central androgen action probably stimulates GnRH release. PMID- 16339201 TI - Dominant role of sarcoendoplasmic reticulum Ca2+-ATPase pump in Ca2+ homeostasis and exocytosis in rat pancreatic beta-cells. AB - The exocytosis of insulin-containing granules from pancreatic beta-cells is tightly regulated by changes in cytosolic Ca2+ concentration ([Ca2+]i). We investigated the role of the sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) pump, Na+/Ca2+ exchanger, and plasma membrane Ca2+-ATPase pump in the Ca2+ dynamics of single rat pancreatic beta-cells. When the membrane potential was voltage clamped at -70 mV (in 3 mm glucose at approximately 22 or 35 C), SERCA pump inhibition dramatically slowed (approximately 4-fold) cytosolic Ca2+ clearance and caused a sustained rise in basal [Ca2+]i via the activation of capacitative Ca2+ entry. SERCA pump inhibition increased ( approximately 1.8-fold) the amplitude of the depolarization-triggered Ca2+ transient at approximately 22 C. Inhibition of the Na+/Ca2+ exchanger or plasma membrane Ca2+-ATPase pump had only minor effects on Ca2+ dynamics. Simultaneous measurement of [Ca2+]i and exocytosis (with capacitance measurement) revealed that SERCA pump inhibition increased the magnitude of depolarization-triggered exocytosis. This enhancement in exocytosis was not due to the slowing of the cytosolic Ca2+ clearance but was closely correlated to the increase in the peak of the depolarization-triggered Ca2+ transient. When compared at similar [Ca2+]i with controls, the rise in basal [Ca2+]i during SERCA pump inhibition did not cause any enhancement in the magnitude of the ensuing depolarization-triggered exocytosis. Therefore, we conclude that in rat pancreatic beta-cells, the rapid uptake of Ca2+ by SERCA pump limits the peak amplitude of depolarization-triggered [Ca2+]i rise and thus controls the amount of insulin secretion. PMID- 16339202 TI - Ghrelin neutralization by a ribonucleic acid-SPM ameliorates obesity in diet induced obese mice. AB - Ghrelin, an acylated peptide secreted from the stomach, acts as a short-term signal of nutrient depletion. Ghrelin is an endogenous ligand for the GH secretagogue receptor 1a, a G protein-coupled receptor expressed in the hypothalamus and pituitary. We used a synthetic oligonucleotide, NOX-B11-2, capable of specific high-affinity binding to bioactive ghrelin to determine whether ghrelin neutralization would alter indices of energy balance in vivo. This novel type of ghrelin-blocking agent, called an RNA Spiegelmer (SPM), is a polyethylene glycol-modified l-RNA oligonucleotide, the nonnatural configuration of which confers in vivo stability. NOX-B11-2 blocked ghrelin mediated activation of GH secretagogue receptor 1a in cell culture (IC50 approximately 5 nm). We explored the effects of acute NOX-B11-2 administration on ghrelin-induced feeding in mice. NOX-B11-2 (66 mg/kg, sc) blocked ghrelin-induced feeding and was without effect on feeding evoked by an orally active nonpeptide ghrelin receptor agonist. We demonstrated that selective ghrelin blockade effectively promoted weight loss in diet-induced obese (DIO) mice. Chronic infusion of NOX-B11-2 (33 mg/kg.d, sc) to DIO mice evoked body weight loss for 13 d and reduced food intake and fat mass relative to control SPM-infused mice. In a 7-d study, DIO mice infused with NOX B11-2 (33 mg/kg.d, sc) showed body weight loss, compared with animals receiving control SPM. This effect was directly mediated by SPM neutralization of ghrelin because NOX-B11-2 administration to ghrelin-deficient mice resulted in no weight loss. The decreased obesity observed in SPM-treated DIO mice provides validation for ghrelin neutralization as a potential antiobesity therapy. PMID- 16339203 TI - Dedifferentiation of adult monkey Sertoli cells through activation of extracellularly regulated kinase 1/2 induced by heat treatment. AB - Sertoli cells play a key role in triggering and regulating the process of spermatogenesis. Failure of a Sertoli cell to mature functionally will presumably render it incapable of supporting germ cell survival and development that appeared after puberty. Expression of cytokeratin 18 (ck-18) intermediate filaments indicates a state of undifferentiation usually observed in Sertoli cells of prepubertal testis. In this study we demonstrated that local testicular heat treatment of adult monkey with water at 43 C for 30 min once daily for 2 consecutive days was capable of activating reexpression of ck-18 in Sertoli cells, which was coincident with activation of ERK1/2 and Akt kinases. Using primary Sertoli cell culture isolated from adult monkey testis, we further confirmed that the heat treatment of the cells at 43 C could also induce ck-18 reexpression, which was similar to the in vivo treatment. ERK MAPK was also induced by the heat treatment in a time- and protein kinase A (PKA)-dependent manner. After blocking the ERK MAPK signaling pathway, an inhibition of ck-18 expression in the cultured Sertoli cells was observed, and this inhibitory effect was also detected by blocking the PKA activation. However, ck-18 activation in Sertoli cells remained unaltered when the phosphatidylinositol 3-kinase/Akt pathway was blocked. In conclusion, the heat treatment of adult monkey Sertoli cells are capable of inducing a reversible change in the Sertoli cells from an adult differentiated state to an immature-like dedifferentiated state through PKA ERK MAPK-dependent pathways but not via the phosphatidylinositol 3-kinase/Akt pathway. PMID- 16339204 TI - Attenuated insulin release and storage in fetal sheep pancreatic islets with intrauterine growth restriction. AB - We determined in vivo and in vitro pancreatic islet insulin secretion and glucose metabolism in fetuses with intrauterine growth restriction (IUGR) caused by chronic placental insufficiency to identify functional deficits in the fetal pancreas that might be caused by nutrient restriction. Plasma insulin concentrations in the IUGR fetuses were 69% lower at baseline and 76% lower after glucose-stimulated insulin secretion (GSIS). Similar deficits were observed with arginine-stimulated insulin secretion. Fetal islets, immunopositive for insulin and glucagon, secreted insulin in response to increasing glucose and KCl concentrations. Insulin release as a fraction of total insulin content was greater in glucose-stimulated IUGR islets, but the mass of insulin released per IUGR islet was lower because of their 82% lower insulin content. A deficiency in islet glucose metabolism was found in the rate of islet glucose oxidation at maximal stimulatory glucose concentrations (11 mmol/liter). Thus, pancreatic islets from nutritionally deprived IUGR fetuses caused by chronic placental insufficiency have impaired insulin secretion caused by reduced glucose stimulated glucose oxidation rates, insulin biosynthesis, and insulin content. This impaired GSIS occurs despite an increased fractional rate of insulin release that results from a greater proportion of releasable insulin as a result of lower insulin stores. Because this animal model recapitulates the human pathology of chronic placental insufficiency and IUGR, the beta-cell GSIS dysfunction in this model might indicate mechanisms that are developmentally adaptive for fetal survival but in later life might predispose offspring to adult-onset diabetes that has been previously associated with IUGR. PMID- 16339205 TI - Three-dimensional organization of thyroid cells into follicle structures is a pivotal factor in the control of sodium/iodide symporter expression. AB - Expression of sodium/iodide symporter (NIS) by thyroid epithelial cells is primarily regulated by TSH, which acts at the level of NIS gene transcription. Knowledge of the mechanisms governing NIS expression mainly comes from studies of rat thyroid-derived cell lines forming cell monolayers. In this study we investigated the impact of the three-dimensional organization of thyroid cells into follicles on the regulation of NIS expression. We used porcine thyrocytes in primary culture that, depending on cell density and the moment TSH is added, either predominantly form a cell monolayer (CM) or reconstitute thyroid follicles (RTF). NIS expression analyzed at transcript and protein levels was remarkably high in RTF compared with CM. Cells forming RTF were NIS positive, whereas in CM, NIS was only detected in the limited number of cells forming follicle-like structures. When thyrocytes were cultured at increasing cell density to obtain a gradual shift from CM to RTF, the progressive increase in the proportion of cells enrolled in RTF was accompanied by a parallel increase in NIS expression. Other TSH-regulated genes, thyroperoxidase, Na(+),K(+)-adenosine triphosphatase alpha subunit, and thyroglobulin, were expressed at similar levels whatever the organization of thyrocytes in culture. The transcription factor, Pax-8, was equally expressed in NIS-negative CM and NIS-positive RTF. We show that TSH highly activates NIS expression only when thyrocytes have undergone histiotypic morphogenesis. This finding suggests that TSH activation of NIS gene transcription might involve, in addition to Pax-8, a regulatory factor(s) whose synthesis and/or activity are triggered by cell-cell interaction(s) occurring in the course of folliculogenesis. PMID- 16339206 TI - Steroid receptor coactivator-1-deficient mice exhibit altered hypothalamic pituitary-adrenal axis function. AB - Steroidogenic factor-1 (SF-1), has emerged as a critical nuclear receptor regulating development and differentiation at several levels of the hypothalamic pituitary-steroidogenic axis. Although many coregulatory factors have been shown to physically and functionally interact with SF-1, the relative importance of these interactions in SF-1 target tissues has not been thoroughly established. In this study we assessed roles of steroid receptor coactivator-1 (SRC-1) in hypothalamic-pituitary-adrenal (HPA) axis function using SRC-1-deficient (SRC-1-/ ) mice in the absence or presence of SF-1 haploinsufficiency. Surprisingly, SRC-1 deficiency did not alter baseline HPA axis function or the acute rise in corticosterone after ACTH administration and failed to exacerbate adrenocortical dysfunction in SF-1+/- mice. However, after exposure to paradigms of acute and chronic stress, SRC-1-/- mice exhibited an elevation in serum corticosterone despite normal (nonsuppressed) ACTH, suggesting an increase in adrenal sensitivity as well as a concomitant defect in glucocorticoid-mediated feedback inhibition of the HPA axis. An examination of potential compensatory mechanism(s) revealed an increase in adrenal weight, selective elevation of melanocortin 2 receptor mRNA, and a coincident increase in SRC-2 and SRC-3 expression in SRC-1-/ adrenals. A reduction in blood glucose was observed in SRC-1-/- mice after chronic stress, consistent with a generalized state of glucocorticoid resistance. Dexamethasone suppression tests confirmed a weakened ability of glucocorticoids to 1) elevate serum glucose levels and induce hepatic phosphoenolpyruvate carboxykinase transcription and 2) suppress pituitary proopiomelanocortin transcript levels in SRC-1-/- animals. Collectively, these data are consistent with an indispensable role for SRC-1 in mediating actions of glucocorticoids in pituitary and liver. PMID- 16339207 TI - Liver triglyceride secretion and lipid oxidative metabolism are rapidly altered by leptin in vivo. AB - Leptin has potent lipid-lowering effects in peripheral tissues and plasma that are proposed to be important for the prevention of cellular lipotoxicity and insulin resistance. The current study addressed in vivo the effects of acute leptin delivery on liver triglyceride (TG) metabolism, the consequence of hepatic leptin action on whole-body TG homeostasis, and the mechanisms of leptin action. A 120-min iv leptin infusion (plasma leptin, approximately 14 ng/ml) decreased liver TG levels (53 +/- 3%; P = 0.001), but not skeletal muscle TG levels, and increased liver phosphatidylinositol 3-kinase activity (341 +/- 95%; P = 0.01) in lean rats. Leptin had no effect on liver TG levels or phosphatidylinositol 3 kinase activity in diet-induced obese rats. In lean animals, leptin decreased the plasma TG concentration (20 +/- 7%; P = 0.017), the rate of TG accumulation in plasma after tyloxapol administration (26 +/- 6%; P = 0.003), and TG secretion from isolated liver (51 +/- 8%; P = 0.004). To determine possible metabolic fates of depleted hepatic TG, we assessed leptin effects on liver oxidative metabolism. Leptin increased hepatic acetyl-coenzyme A carboxylase phosphorylation (85 +/- 13%; P = 0.006), fatty acid oxidation (49 +/- 7%; P = 0.001) and ketogenesis (69 +/- 15%; P = 0.004). Finally, intracerebroventricular delivery of leptin for 120 min had no effect on liver TG levels, but did increase signal transducer and activator of transcription 3 phosphorylation (162 +/- 40%; P = 0.02). These data present in vivo evidence for a role for leptin in the acute regulation of hepatic TG metabolism, and whole body TG homeostasis. A likely contributing mechanism for these effects is leptin-induced partitioning of TG into oxidative pathways. PMID- 16339208 TI - Maternal ghrelin plays an important role in rat fetal development during pregnancy. AB - Ghrelin, an acylated peptide serving as an endogenous ligand for GH secretagogue receptor (GHS-R), was originally isolated from rat and human stomach. In this study, we report the critical role of maternal ghrelin in fetal development. High levels of ghrelin receptor (GHS-R) mRNA were detected in various peripheral fetal tissues beginning at embryonic d 14 and lasting until birth. Fetal GHS-R expression was also confirmed in fetal tissues by immunohistochemistry. Autoradiography revealed that both des-acyl ghrelin and acyl ghrelin bind to fetal tissues. Chronic treatment of mothers with ghrelin resulted in a significant increase in birth weight in comparison to newborns from saline treated mothers. Even when maternal food intake after ghrelin treatment was restricted through paired feeding, significant stimulation of fetal development still occurred. Conversely, active immunization of mothers against ghrelin decreased fetal body weight during pregnancy. A single ghrelin injection into the mother increased circulating ghrelin levels in the fetus within 5 min of injection, suggesting that maternal ghrelin transits easily to the fetal circulation. High levels of des-acyl ghrelin were detected in fetal blood and amniotic fluid. Both acylated and des-acyl ghrelin increased [3H]thymidine and 5 bromo-2'-deoxyuridine incorporation of cultured fetal skin cells in a dose dependent manner, and calcium-imaging analysis revealed that acyl and des-acyl ghrelin increased the Ca2+ influx in discrete cultured fetal skin cells, respectively. These results indicate that maternal ghrelin regulates fetal development during the late stages of pregnancy. PMID- 16339209 TI - Tracing from fat tissue, liver, and pancreas: a neuroanatomical framework for the role of the brain in type 2 diabetes. AB - The hypothalamus uses hormones and the autonomic nervous system to balance energy fluxes in the body. Here we show that the autonomic nervous system has a distinct organization in different body compartments. The same neurons control intraabdominal organs (intraabdominal fat, liver, and pancreas), whereas sc adipose tissue located outside the abdominal compartment receives input from another set of autonomic neurons. This differentiation persists up to preautonomic neurons in the hypothalamus, including the biological clock, that have a distinct organization depending on the body compartment they command. Moreover, we demonstrate a neuronal feedback from adipose tissue that reaches the brainstem. We propose that this compartment-specific organization offers a neuroanatomical perspective for the regional malfunction of organs in type 2 diabetes, where increased insulin secretion by the pancreas and disturbed glucose metabolism in the liver coincide with an augmented metabolic activity of visceral compared with sc adipose tissue. PMID- 16339210 TI - Testis structure and function in a nongenetic hyperadipose rat model at prepubertal and adult ages. AB - There are few data for hormonal levels and testis structure and function during postnatal development in rats neonatally treated with monosodium L-glutamate (MSG). In our study, newborn male pups were ip injected with MSG (4 mg/g body weight) every 2 d up to 10 d of age and investigated at prepubertal and adult ages. Plasma levels of leptin, LH, FSH, prolactin, testosterone (T), corticosterone, and free T4 (FT4) were measured. MSG rats displayed elevated circulating levels of corticosterone and hyperadiposity/hyperleptinemia, regardless of the age examined; conversely, circulating prolactin levels were not affected. Moreover, prepubertal MSG rats revealed a significant (P < 0.05) reduction in testis weight and the number of Sertoli (SC) and Leydig cells per testis. Leptin plasma levels were severalfold higher (2.41 vs. 8.07; P < 0.05) in prepubertal MSG rats, and these animals displayed plasma LH, FSH, T, and FT4 levels significantly decreased (P < 0.05). Taken together, these data indicate that testis development, as well as SC and Leydig cell proliferation, were disturbed in prepubertal MSG rats. Adult MSG rats also displayed significantly higher leptin plasma levels (7.26 vs. 27.04; P < 0.05) and lower (P < 0.05) LH and FSH plasma levels. However, T and FT4 plasma levels were normal, and no apparent alterations were observed in testis structure of MSG rats. Only the number of SCs per testis was significantly (P < 0.05) reduced in the adult MSG rats. In conclusion, although early installed hyperadipose/hyperleptinemia phenotype was probably responsible for the reproductive axis damages in MSG animals, it remains to be investigated whether this condition is the main factor for hypothalamus-pituitary-gonadal axis dysfunction in MSG rats. PMID- 16339211 TI - Activation of the Y1 receptor by neuropeptide Y regulates the growth of prostate cancer cells. AB - This study deals with the role of neuropeptide Y (NPY) in the regulation of cell proliferation. NPY is expressed in the normal and tumoral prostate, but no data on its possible role in prostate cancer (PCa) progression are available. Therefore, we evaluated the direct effect of NPY on the growth of the human PCa cell lines LNCaP (androgen dependent) and DU145 and PC3 (androgen independent). All PCa cell lines expressed Y1-R gene and protein. NPY treatment reduced the proliferation of LNCaP and DU145 cells and increased that of PC3 cells. The Y1-R antagonist BIBP3226 abolished such effects, suggesting a mandatory role of Y1-R in this process. LNCaP cells showed elevated constitutive levels of phosphorylated ERK1/2, which were not affected by NPY. In DU145 cells, NPY stimulated a long-lasting ERK1/2 activation, whereas, in PC3 cells, this effect was rapid and transient and required activation of protein kinase C. Moreover, in both cell lines, pretreatment with BIBP3226 prevented the NPY-induced ERK1/2 phosphorylation, further supporting Y1-R involvement. NPY treatment reduced forskolin-stimulated cAMP accumulation only in PC3 cells and did not change intracellular calcium concentration in any PCa cell line. These data indicate that NPY may directly regulate PCa cell growth via Y1-R. The direction of this effect appears to be related to the time kinetics of MAPK activation, i.e. long lasting vs. transient, and to the clone-specific involvement of other intracellular signals. These findings suggest that NPY-related mechanisms might play a relevant role in the progression of PCa, at both androgen dependent and independent stages. PMID- 16339212 TI - Emergencies in hereditary haemorrhagic telangiectasia. AB - BACKGROUND: Hereditary haemorrhagic telangiectasia (HHT) is a systemic autosomal dominant vascular disease. Although the clinical picture is that of a chronic disabling disease, vascular malformations can suddenly lead to life-threatening conditions. AIM: To assess the frequency and type of emergency acute complications in HHT. DESIGN: Retrospective case-note review. METHODS: From August 2000 to December 2004, our specialized HHT centre saw 139 patients (74 males, 65 females, mean age 45.5 years, range 14-77) with a definite diagnosis of HHT. We reviewed their clinical files and recorded all visits for acute complications (massive nosebleeds, haematemesis, melaena, haematochezia, haemothorax, haemoptysis, TIA/ischaemic stroke, haemorrhagic stroke, brain abscess). RESULTS: Fifty patients (35.9%) had at least one acute complication. There were a total of 93 visits potentially involving the emergency department. Most commonly, patients sought urgent medical attention for nosebleeds and gastrointestinal bleeding (63.4%), but there were also disorders of the brain, lung, heart and liver. DISCUSSION: Acute complications of HHT are not uncommon and can be severe and wide-ranging. Physicians should be aware of HHT and its major complications, as a prompt diagnosis is essential to direct patients to the most appropriate therapies, and to suggest screening for visceral involvement in their relatives. PMID- 16339213 TI - Spastin and atlastin, two proteins mutated in autosomal-dominant hereditary spastic paraplegia, are binding partners. AB - The pure hereditary spastic paraplegias (HSPs) are a group of conditions in which there is a progressive length-dependent degeneration of the distal ends of the corticospinal tract axons, resulting in spastic paralysis of the legs. Pure HSPs are most frequently inherited in an autosomal-dominant pattern and are commonly caused by mutations either in the SPG4 gene spastin or in the SPG3A gene atlastin. To identify binding partners for spastin, we carried out a yeast two hybrid screen on a brain cDNA library, using spastin as bait. Remarkably, nearly all of the positive interacting prey clones coded for atlastin. We have verified the physiological relevance of this interaction using co-immunoprecipitation, glutathione S-transferase pull-down and intracellular co-localization experiments. We show that the spastin domain required for binding to atlastin lies within the N-terminal 80 residues of the protein, a region that is only present in the predominantly cytoplasmic, full-length spastin isoform. These data suggest that spastin and atlastin function in the same biochemical pathway and that it is the cytoplasmic function of spastin which is important for the pathogenesis of HSP. They also provide further evidence for a physiological and pathological role of spastin in membrane dynamics. PMID- 16339214 TI - Transplanted ALDHhiSSClo neural stem cells generate motor neurons and delay disease progression of nmd mice, an animal model of SMARD1. AB - Spinal muscular atrophy with respiratory distress type 1 (SMARD1) is an infantile autosomal-recessive motor neuron disease caused by mutations in the immunoglobulin micro-binding protein 2. We investigated the potential of a spinal cord neural stem cell population isolated on the basis of aldehyde dehydrogenase (ALDH) activity to modify disease progression of nmd mice, an animal model of SMARD1. ALDH(hi)SSC(lo) stem cells are self-renewing and multipotent and when intrathecally transplanted in nmd mice generate motor neurons properly localized in the spinal cord ventral horns. Transplanted nmd animals presented delayed disease progression, sparing of motor neurons and ventral root axons and increased lifespan. To further investigate the molecular events responsible for these differences, microarray and real-time reverse transcription-polymerase chain reaction analyses of wild-type, mutated and transplanted nmd spinal cord were undertaken. We demonstrated a down-regulation of genes involved in excitatory amino acid toxicity and oxidative stress handling, as well as an up regulation of genes related to the chromatin organization in nmd compared with wild-type mice, suggesting that they may play a role in SMARD1 pathogenesis. Spinal cord of nmd-transplanted mice expressed high transcript levels for genes related to neurogenesis such as doublecortin (DCX), LIS1 and drebrin. The presence of DCX-expressing cells in adult nmd spinal cord suggests that both exogenous and endogenous neurogeneses may contribute to the observed nmd phenotype amelioration. PMID- 16339216 TI - Hamartin, the tuberous sclerosis complex 1 gene product, interacts with polo-like kinase 1 in a phosphorylation-dependent manner. AB - Tuberous sclerosis complex (TSC) is a tumor suppressor gene syndrome caused by mutations in TSC1 and TSC2. Hamartin and tuberin, the products of TSC1 and TSC2, respectively, form heterodimers and inhibit the mammalian target of rapamycin. Previously, we have shown that hamartin is phosphorylated by CDC2/cyclin B1 during the G(2)/M phase of the cell cycle. Here, we report that hamartin is localized to the centrosome and that phosphorylated hamartin and phosphorylated tuberin co-immunoprecipitate with the mitotic kinase Plk1. Plk1 interacts with the N-terminus of hamartin (amino acids 1-880), which contains two potential Plk1 binding sites (T310 and S332). Phosphorylated hamartin interacts with Plk1 independent of tuberin with all three proteins present in a complex. A non phosphorylatable hamartin mutant with an alanine substitution at residue T310 does not interact with Plk1, whereas a non-phosphorylatable hamartin mutant at residue S332 in conjunction with alanine mutations at the other CDC2/cyclin B1 sites (T417, S584 and T1047) does not impact hamartin binding to Plk1. Hamartin negatively regulates the protein levels of Plk1. Finally, Tsc1(-/-) mouse embryonic fibroblasts (MEFs) have increased number of centrosomes and increased DNA content, compared to Tsc1(+/+) cells. Both phenotypes are rescued after pre treatment with the mTOR inhibitor rapamycin. RNAi inhibition of Plk1 in Tsc1(-/-) MEFs failed to rescue the increased centrosome number phenotype. These data reveal a novel subcellular localization for hamartin and a novel interaction partner for the hamartin/tuberin complex and implicate hamartin and mTOR in the regulation of centrosome duplication. PMID- 16339215 TI - Gain-of-function haplotypes in the vesicular monoamine transporter promoter are protective for Parkinson disease in women. AB - The vesicular monoamine transporter can protect against toxins that induce an acute parkinsonian syndrome. It has been hypothesized that cytoplasmic dopamine has subacute toxic effects in Parkinson Disease (PD) leading to neuronal death and clinical symptoms. Regulatory polymorphisms in the brain form of the vesicular monoamine transporter (VMAT2) which affect its quantitative expression might therefore serve as genetic risk factors for PD. We have screened the promoter region of the gene for VMAT2 (SLC18A2) and identified several novel polymorphisms that form discrete haplotypes. We have tested the common halpotypes in SLC18A2 for functional effects in reporter gene assays and found that there are several gain-of-function haplotypes that display significantly increased transcriptional activity from the reference element. These gain-of-function haplotypes were tested for association with PD and found to confer a protective effect that was selective for females. This finding is consistent with the prediction that increased sequestration of dopamine in secretory vesicles by VMAT2 is protective for PD. PMID- 16339217 TI - Economic evaluation of a general practitioner with special interests led dermatology service in primary care. AB - OBJECTIVE: To carry out an economic evaluation of a general practitioner with special interest service for non-urgent skin problems compared with hospital outpatient care. DESIGN: Cost effectiveness analysis and cost consequences analysis alongside a randomised controlled trial. SETTING: General practitioner with special interest dermatology service covering 29 general practices in Bristol. PARTICIPANTS: Adults referred to a hospital dermatology clinic who were potentially suitable for management by a general practitioner with special interest. INTERVENTIONS: Participants were randomised 2:1 to receive either care by general practitioner with special interest service or usual hospital outpatient care. MAIN OUTCOME MEASURES: Costs to NHS, patients, and companions, and costs of lost production. Cost effectiveness, using the two primary outcomes of dermatology life quality index scores and improved patient perceived access, was assessed by incremental cost effectiveness ratios and cost effectiveness acceptability curves. Cost consequences are presented in relation to all costs and both primary and secondary outcomes from the trial. RESULTS: Costs to the NHS for patients attending the general practitioner with special interest service were 208 pounds sterling (361 dollars; 308 euro) compared with 118 pounds sterling for hospital outpatient care. Based on analysis with imputation of missing data, costs to patients and companions were 48 pounds sterling and 51 pounds sterling, respectively; costs of lost production were 27 pounds sterling and 34 pounds sterling, respectively. The incremental cost effectiveness ratios for general practitioner with special interest care over outpatient care were 540 pounds sterling per one point gain in the dermatology life quality index and 66 pounds sterling per 10 point change in the access scale. CONCLUSIONS: The general practitioner with special interest service for dermatology is more costly than hospital outpatient care, but this additional cost needs to be weighed against improved access and broadly similar health outcomes. PMID- 16339218 TI - General practitioners with special interests--not a cheap option. PMID- 16339219 TI - Risk factors for pulmonary tuberculosis in Russia: case-control study. AB - OBJECTIVES: To determine risk factors for pulmonary tuberculosis in Russia. DESIGN: Case-control study of exposure to a variety of risk factors before and during the development of pulmonary tuberculosis. SETTING: Large city in Russia. PARTICIPANTS: Cases were 334 consecutive adults diagnosed as having culture confirmed pulmonary tuberculosis between 1 January 2003 and 31 December 2003. Controls were 334 individuals sampled from a validated population registry, matched for age and sex to the patients with tuberculosis. A questionnaire collected information on potential risk factors. MAIN OUTCOME MEASURES: Risk factors associated with the development of tuberculosis. RESULTS: The main risk factors for tuberculosis were low accumulated wealth (univariate odds ratio 16.70), financial insecurity (5.67), consumption of unpasteurised milk (3.58), diabetes (2.66), living with a relative with tuberculosis (2.94), being unemployed (6.10), living in overcrowded conditions (2.99), illicit drug use (8.74), and a history of incarceration in both pretrial detention centres (5.70) and prison (12.50). CONCLUSIONS: When prevalence of exposure is taken into account the most important factors in the development of pulmonary tuberculosis in Russia are exposure to raw milk and unemployment. PMID- 16339221 TI - Facial transplantation. PMID- 16339222 TI - Extra scrutiny for industry funded trials. PMID- 16339220 TI - Randomised placebo controlled multicentre trial to assess short term clarithromycin for patients with stable coronary heart disease: CLARICOR trial. AB - OBJECTIVE: To determine if the macrolide clarithromycin affects mortality and cardiovascular morbidity in patients with stable coronary heart disease. DESIGN: Centrally randomised multicentre trial. All parties at all stages were blinded. Analyses were by intention to treat. SETTING: Five Copenhagen University cardiology departments and a coordinating centre. PARTICIPANTS: 13,702 patients aged 18 to 85 years who had a discharge diagnosis of myocardial infarction or angina pectoris in 1993-9 and alive in August 1999 were invited by letter; 4373 were randomised. INTERVENTIONS: Two weeks' treatment with clarithromycin 500 mg/day or matching placebo. MAIN OUTCOME MEASURES: PRIMARY OUTCOME: composite of all cause mortality, myocardial infarction, or unstable angina pectoris during three years' follow-up. Secondary outcome: composite of cardiovascular mortality, myocardial infarction, or unstable angina pectoris. The outcomes were obtained from Danish registers and were blindly assessed by the event committee. RESULTS: 2172 participants were randomised to clarithromycin and 2201 to placebo. We found no significant effects of clarithromycin on the primary outcome (hazard ratio 1.15, 95% confidence interval 0.99 to 1.34) or secondary outcome (1.17, 0.98 to 1.40). Mortality was significantly higher in the clarithromycin arm (1.27, 1.03 to 1.54; P = 0.03) as a result of significantly higher cardiovascular mortality (1.45, 1.09 to 1.92; P = 0.01). CONCLUSIONS: Short term clarithromycin in patients with stable coronary heart disease may cause significantly higher cardiovascular mortality. The long term safety of clarithromycin in patients with stable ischaemic heart disease should be examined. Trial registration ClinicalTrials.gov NCT00121550. PMID- 16339223 TI - Sharing the benefits of genetic research. PMID- 16339224 TI - Is methadone too dangerous for opiate addiction? PMID- 16339225 TI - Treating non-competent patients. PMID- 16339230 TI - Israelis turn to timer device to facilitate passive euthanasia. PMID- 16339231 TI - The Netherlands regulates ending the lives of severely ill neonates. PMID- 16339232 TI - Yemen embarks on its seventh round of polio immunisations. PMID- 16339237 TI - Drug companies told that sponsoring patients' groups might help win approval for their products. PMID- 16339240 TI - Inquiry needed to discover why black people are more likely to be admitted to mental hospitals than white people. PMID- 16339243 TI - Older people are wrongly excluded from drug trials. PMID- 16339246 TI - Romania's policy of emptying its orphanages raises controversy. PMID- 16339247 TI - The ageing population of the United Kingdom and cardiovascular disease. PMID- 16339249 TI - Functional limitation in long standing illness and quality of life: evidence from a national survey. PMID- 16339248 TI - Payment to healthcare professionals for patient recruitment to trials: a systematic review. PMID- 16339250 TI - Developing guidelines for medical students about the examination of patients under 18 years old. PMID- 16339251 TI - Treating obesity in individuals and populations. PMID- 16339252 TI - eHealth and the future: promise or peril? PMID- 16339253 TI - Rise and demise of the hospital: a reappraisal of nursing. PMID- 16339254 TI - Practice based commissioning: applying the research evidence. PMID- 16339255 TI - Primary care in the United States: problems and possibilities. PMID- 16339256 TI - Designing research in vulnerable populations: lessons from HIV prevention trials that stopped early. PMID- 16339257 TI - Human and animal health: strengthening the link: politics and economics inhibited control of anthrax last century. PMID- 16339258 TI - Human and animal health: strengthening the link: methodological concerns about animal facilitated therapy with dolphins. PMID- 16339259 TI - Role of vaccinating poultry against avian flu. PMID- 16339261 TI - Human and animal health: strengthening the link: child and animal abuse are linked. PMID- 16339262 TI - Inequalities and christmas yet to come. PMID- 16339263 TI - Committee on Radioactive Waste Management lacks medical input. PMID- 16339264 TI - New ABPI code provides more information for patients. PMID- 16339265 TI - The sniff as a unit of olfactory processing. AB - Sniffing is a rhythmic motor process essential for the acquisition of olfactory information. Recent behavioral experiments show that using a single sniff rats can accurately discriminate between very similar odors and fail to improve their accuracy by taking multiple sniffs. This implies that each sniff has the potential to provide a complete snapshot of the local olfactory environment. The discrete and intermittent nature of sniffing has implications beyond the physical process of odor capture as it strongly shapes the flow of information into the olfactory system. We review electrophysiological studies-primarily from anesthetized rodents-demonstrating that olfactory neural responses are coupled to respiration. Hence, the "sniff cycle" might play a role in odor coding, by allowing the timing of spikes with respect to the phase of the respiration cycle to encode information about odor identity or concentration. We also discuss behavioral and physiological results indicating that sniffing can be dynamically coordinated with other rhythmic behaviors, such as whisking, as well as with rhythmic neural activity, such as hippocampal theta oscillations. Thus, the sniff cycle might also facilitate the coordination of the olfactory system with other brain areas. These converging lines of empirical data support the notion that each sniff is a unit of olfactory processing relevant for both neural coding and inter-areal coordination. Further electrophysiological recordings in behaving animals will be necessary to assess these proposals. PMID- 16339266 TI - Anatomical contributions to odorant sampling and representation in rodents: zoning in on sniffing behavior. AB - Odorant sampling behaviors such as sniffing bring odorant molecules into contact with olfactory receptor neurons (ORNs) to initiate the sensory mechanisms of olfaction. In rodents, inspiratory airflow through the nose is structured and laminar; consequently, the spatial distribution of adsorbed odorant molecules during inspiration is predictable. Physicochemical properties such as water solubility and volatility, collectively called sorptiveness, interact with behaviorally regulable variables such as inspiratory flow rate to determine the pattern of odorant deposition along the inspiratory path. Populations of ORNs expressing the same odorant receptor are distributed in strictly delimited regions along this inspiratory path, enabling different deposition patterns of the same odorant to evoke different patterns of neuronal activation across the olfactory epithelium and in the olfactory bulb. We propose that both odorant sorptive properties and the regulation of sniffing behavior may contribute to rodents' olfactory capacities by this mechanism. In particular, we suggest that the motor regulation of sniffing behavior is substantially utilized for purposes of "zonation" or the direction of odorant molecules to defined intranasal regions and hence toward distinct populations of receptor neurons, pursuant to animals' sensory goals. PMID- 16339268 TI - The sniff is part of the olfactory percept. AB - In this review, we use data obtained primarily from humans to argue that sniffs are not merely a stimulus carrier but are rather a central component of the olfactory percept. We argue that sniffs 1) are necessary for the olfactory percept, 2) affect odorant intensity perception and identity perception, 3) drive activity in olfactory cortex, 4) are rapidly modulated in an odorant-dependent fashion by a dedicated olfactomotor system, and 5) are sufficient to generate an olfactory percept of some sort even in the absence of odor. PMID- 16339269 TI - Winging it: moth flight behavior and responses of olfactory neurons are shaped by pheromone plume dynamics. AB - Terrestrial odor plumes have a physical structure that results from turbulence in the fluid environment. The rapidity of insect flight maneuvers within a plume indicates that their responses are dictated by fleeting (<1 s) rather than longer (>1 s) exposures to odor imposed by physical variables that distribute odor molecules in time and space. Even though encounters with pheromone filaments are brief, male moths responding to female-produced pheromones are remarkably able to extract information relating to the biological properties of these olfactory signals. These properties include the types of molecule present and their relative abundances. Thus, peripheral and central olfactory neurons are capable of representing these biological properties of a pheromone plume within the context of a temporally irregular and unpredictable signal. The mechanisms underlying olfactory processing of these signals with respect to their biological and physical properties are discussed in the context of a behavioral framework. PMID- 16339270 TI - Respiratory modulation of olfactory neurons in the rodent brain. AB - In this review we report data from freely breathing animals in an attempt to show how respiratory dynamics can influence bulbar and cortical activity. Relying on in vivo data as well as in vitro observations, we try to emphasize the multiple mechanisms that underlie this modulation, its multiple origins, and its possible functional role. PMID- 16339271 TI - The fluid mechanics of arthropod sniffing in turbulent odor plumes. AB - Many arthropods capture odorant molecules from the environment using antennae or antennules bearing arrays of chemosensory hairs. The penetration of odorant carrying water or air into the spaces between these chemosensory hairs depends on the speed at which they are moved through the surrounding fluid. Therefore, antennule flicking by crustaceans and wing fanning by insects can have a profound impact on the odorant encounter rates of the chemosensory sensilla they bear; flicking and fanning are examples of sniffing. Odors are dispersed in the environment by turbulent wind or water currents. On the scale of an antenna or antennule, an odor plume is not a diffuse cloud but rather is a series of fine filaments of scent swirling in odor-free water. The spatiotemporal pattern of these filaments depends on distance from the odor source. The physical interaction of a hair-bearing arthropod antennule with the surrounding fluid affects the temporal patterns of odor concentration an animal intercepts when it sniffs in a turbulent odor plume. PMID- 16339272 TI - Overexpression of Kruppel-like factor 7 regulates adipocytokine gene expressions in human adipocytes and inhibits glucose-induced insulin secretion in pancreatic beta-cell line. AB - We have identified Kruppel-like factor 7 (KLF7) as a new candidate for conferring susceptibility to type 2 diabetes. To ascertain the possible involvement of KLF7 in the pathogenesis of type 2 diabetes, we examined the functional roles of KLF7 in various types of cells. In human adipocytes overexpressing KLF7, the expression of adiponectin and leptin was decreased compared with that in control cells, whereas expression of IL-6 was increased. In the insulin-secreting cell line (HIT-T15 cells), the expression and glucose-induced secretion of insulin were significantly suppressed in KLF7-overexpressed cells compared with control cells, accompanied by the reduction in the expression of glucose transporter 2, sulfonylurea receptor 1, Kir6.2, and pancreatic-duodenal homeobox factor 1. We also found that the overexpression of KLF7 resulted in the decrease of hexokinase 2 expression in smooth muscle cells, and of glucose transporter 2 expression in the HepG2 cells. These results suggest that KLF7 may contribute to the pathogenesis of type 2 diabetes through an impairment of insulin biosynthesis and secretion in pancreatic beta-cells and a reduction of insulin sensitivity in peripheral tissues. Therefore, we suggest that KLF7 plays an important role in the pathogenesis of type 2 diabetes, and may be a useful target for new drugs to aid in the prevention and treatment of this disease. PMID- 16339273 TI - Prolactin receptor signaling mediates the osmotic response of embryonic zebrafish lactotrophs. AB - The pituitary hormone prolactin (PRL) regulates salt and water homeostasis by altering ion retention and water uptake through peripheral osmoregulatory organs. To understand the role of osmotic homeostasis in the development of PRL-secreting lactotrophs, we generated germline transgenic zebrafish coexpressing red fluorescent protein directed by Prolactin regulatory elements (PRL-RFP) and green fluorescent protein by the Pro-opiomelanocortin promoter (POMC-GFP). Transparent embryos expressing fluorescent markers specifically targeted to lactotrophs and corticotrophs, the two pituitary lineages involved in teleost osmotic adaptation, allowed in vivo dynamic tracing of pituitary ontogeny during altered environmental salinity. Physiological osmotic changes selectively regulate lactotroph but not corticotroph proliferation during early ontogeny. These changes are not suppressed by pharmacological dopamine receptor blockade but are completely abrogated by morpholino knockdown of the PRL receptor. PRL receptor signaling exerts robust effects on lactotroph development and plays a permissive role in lactotroph osmo-responsiveness, reflecting the dual peripheral and central interactions required for early pituitary development and embryonic homeostasis. PMID- 16339274 TI - Sequence-specific deoxyribonucleic acid (DNA) recognition by steroidogenic factor 1: a helix at the carboxy terminus of the DNA binding domain is necessary for complex stability. AB - Steroidogenic factor 1 (SF1) is a member of the NR5A subfamily of nuclear hormone receptors and is considered a master regulator of reproduction because it regulates a number of genes encoding reproductive hormones and enzymes involved in steroid hormone biosynthesis. Like other NR5A members, SF1 harbors a highly conserved approximately 30-residue segment called the FTZ-F1 box C-terminal to the core DNA binding domain (DBD) common to all nuclear receptors and binds to 9 bp DNA sequences as a monomer. Here we describe the solution structure of the SF1 DBD in complex with an atypical sequence in the proximal promoter region of the inhibin-alpha gene that encodes a subunit of a reproductive hormone. SF1 forms a specific complex with the DNA through a bipartite motif binding to the major and minor grooves through the core DBD and the N-terminal segment of the FTZ-F1 box, respectively, in a manner previously described for two other monomeric receptors, nerve growth factor-induced-B and estrogen-related receptor 2. However, unlike these receptors, SF1 harbors a helix in the C-terminal segment of the FTZ-F1 box that interacts with both the core DBD and DNA and serves as an important determinant of stability of the complex. We propose that the FTZ-F1 helix along with the core DBD serves as a platform for interactions with coactivators and other DNA-bound factors in the vicinity. PMID- 16339276 TI - Repulsive separation of the cytoplasmic ends of transmembrane helices 3 and 6 is linked to receptor activation in a novel thyrotropin receptor mutant (M626I). AB - Ligand-dependent activation of G protein-coupled receptors (GPCRs) involves repositioning of the juxtacytoplasmic ends of transmembrane helices TM3 and TM6. This concept, inferred from site-directed spin labeling studies, is supported by chemical cross-linking of the cytoplasmic ends of TM3 and TM6 blocking GPCR activation. Here we report a novel constitutive active mutation (M626I) in TM6 of the TSH receptor (TSHR), identified in affected members of a family with nonautoimmune hyperthyroidism. The specific constitutive activity of M626I, measured by its basal cAMP generation corrected for cell surface expression, was 13-fold higher than that of wild-type TSHR. Homology modeling of the TSHR serpentine domain based on the rhodopsin crystal structure suggests that M626 faces the side chain of I515 of TM3 near the membrane-cytoplasmic junction. Steric hindrance of the introduced isoleucine by I515 is consistent with the fact that shorter or more flexible side chains at position 626 did not increase constitutivity. Furthermore, a reciprocal mutation at position 515 (I515M), when introduced into the M626I background, acts as revertant mutation by allowing accommodation of the isoleucine sidechain at position 626 and fully restoring the constitutive activity to the level of wild-type TSHR. Thus, repulsive separation of the juxtacytoplasmic TM6 and TM3 in the M626I model conclusively demonstrates a direct link between the opening of this cytoplasmic face of the receptor structure and G protein coupling. PMID- 16339275 TI - Mechanisms of ligand binding to the parathyroid hormone (PTH)/PTH-related protein receptor: selectivity of a modified PTH(1-15) radioligand for GalphaS-coupled receptor conformations. AB - Mechanisms of ligand binding to the PTH/PTHrP receptor (PTHR) were explored using PTH fragment analogs as radioligands in binding assays. In particular, the modified amino-terminal fragment analog, (125)I [Aib(1,3),Nle8,Gln10,homoarginine11,Ala12,Trp14,Tyr15]rPTH(1-15)NH2, (125)I [Aib(1,3),M]PTH(1-15), was used as a radioligand that we hypothesized to bind solely to the juxtamembrane (J) portion of the PTHR containing the extracellular loops and transmembrane helices. We also employed (125)I-PTH(1-34) as a radioligand that binds to both the amino-terminal extracellular (N) and J domains of the PTHR. Binding was examined in membranes derived from cells expressing either wild-type or mutant PTHRs. We found that the binding of (125)I [Aib(1,3),M]PTH(1-15) to the wild-type PTHR was strongly (approximately 90%) inhibited by guanosine 5'-O-(3-thio)triphosphate (GTPgammaS), whereas the binding of (125)I-PTH(1-34) was only mildly (approximately 25%) inhibited by GTPgammaS. Of these two radioligands, only (125)I-[Aib(1,3),M]PTH(1-15) bound to PTHR-delNt, which lacks most of the receptor's N domain, and again this binding was strongly inhibited by GTPgammaS. Binding of (125)I-[Aib(1,3),M]PTH(1-15) to the constitutively active receptor, PTHR-H223R, was only mildly (approximately 20%) inhibited by GTPgammaS, as was the binding of (125)I-PTH(1-34). In membranes prepared from cells lacking Galpha(S) via knockout mutation of Gnas, no binding of (125)I-[Aib(1,3),M]PTH(1-15) was observed, but binding of (125)I [Aib(1,3),M]PTH(1-15) was recovered by virally transducing the cells to heterologously express Galpha(S). (125)I-PTH(1-34) bound to the membranes with or without Galpha(S). The overall findings confirm the hypothesis that (125)I [Aib(1,3),M]PTH(1-15) binds solely to the J domain of the PTHR. They further show that this binding is strongly dependent on coupling of the receptor to Galpha(S) containing heterotrimeric G proteins, whereas the binding of (125)I-PTH(1-34) can occur in the absence of such coupling. Thus, (125)I-[Aib(1,3),M]PTH(1-15) appears to function as a selective probe of Galpha(S)-coupled, active-state PTHR conformations. PMID- 16339277 TI - Regulation of macrophage inflammatory gene expression by the orphan nuclear receptor Nur77. AB - Members of the nuclear hormone receptor superfamily have emerged as important regulators of macrophage gene expression in inflammation and disease. Previous studies have shown that the lipid-activated receptors peroxisomal proliferator activated receptor and liver X receptor inhibit nuclear factor-kappaB (NF-kappaB) signaling and inflammatory gene expression. We recently identified the NR4A subfamily of orphan nuclear receptors (Nur77/NR4A1, Nurr1/NR4A2, and NOR1/NR4A3) as lipopolysaccharide- and NF-kappaB-responsive genes in macrophages. However, the role of these transcription factors in macrophage gene expression is unknown. We demonstrate here that, in contrast to peroxisomal proliferator-activated receptor and liver X receptor, the role of NR4A receptors in macrophages is proinflammatory. Retroviral expression of Nur77 in macrophages leads to the transcriptional activation of multiple genes involved in inflammation, apoptosis, and cell cycle control. One particularly interesting Nur77-responsive gene is the inducible kinase IKKi/IKKepsilon, an important component of the NF-kappaB signaling pathway. The IKKi promoter contains a functional NR4A binding site and is activated by all three NR4A receptors in transient transfection assays. Consistent with the activation of IKKi, expression of Nur77 in macrophages potentiates the induction of inflammatory gene expression in response to lipopolysaccharide. These results identify a new role for NR4A orphan nuclear receptors in the control of macrophage gene expression during inflammation. PMID- 16339278 TI - Chromium activates glucose transporter 4 trafficking and enhances insulin stimulated glucose transport in 3T3-L1 adipocytes via a cholesterol-dependent mechanism. AB - Evidence suggests that chromium supplementation may alleviate symptoms associated with diabetes, such as high blood glucose and lipid abnormalities, yet a molecular mechanism remains unclear. Here, we report that trivalent chromium in the chloride (CrCl3) or picolinate (CrPic) salt forms mobilize the glucose transporter, GLUT4, to the plasma membrane in 3T3-L1 adipocytes. Concomitant with an increase in GLUT4 at the plasma membrane, insulin-stimulated glucose transport was enhanced by chromium treatment. In contrast, the chromium-mobilized pool of transporters was not active in the absence of insulin. Microscopic analysis of an exofacially Myc-tagged enhanced green fluorescent protein-GLUT4 construct revealed that the chromium-induced accumulation of GLUT4-containing vesicles occurred adjacent to the inner cell surface membrane. With insulin these transporters physically incorporated into the plasma membrane. Regulation of GLUT4 translocation by chromium did not involve known insulin signaling proteins such as the insulin receptor, insulin receptor substrate-1, phosphatidylinositol 3-kinase, and Akt. Consistent with a reported effect of chromium on increasing membrane fluidity, we found that chromium treatment decreased plasma membrane cholesterol. Interestingly, cholesterol add-back to the plasma membrane prevented the beneficial effect of chromium on both GLUT4 mobilization and insulin stimulated glucose transport. Furthermore, chromium action was absent in methyl beta-cyclodextrin-pretreated cells already displaying reduced plasma membrane cholesterol and increased GLUT4 translocation. Together, these data reveal a novel mechanism by which chromium may enhance GLUT4 trafficking and insulin stimulated glucose transport. Moreover, these findings at the level of the cell are consistent with in vivo observations of improved glucose tolerance and decreased circulating cholesterol levels after chromium supplementation. PMID- 16339279 TI - Up-regulation of the progesterone receptor (PR)-C isoform in laboring myometrium by activation of nuclear factor-kappaB may contribute to the onset of labor through inhibition of PR function. AB - Progesterone acting via the progesterone receptor (PR) plays a critical role in maintaining uterine quiescence during pregnancy. In the present study, we tested the hypothesis that the transactivating capability of the PR is down-regulated in the myometrium at term by a change in uterine PR isoform ratio resulting from local activation of the nuclear factor (NF)-kappaB pathway. Overexpression of the truncated PR-C isoform in human myometrial cells inhibited PR-B transactivation. Expression of PR isoforms, PR-A, PR-B, and PR-C, was characterized by immunoblotting and quantitative PCR (Q-PCR) in fundal and lower uterine segment myometrium from pregnant women in labor and not in labor and in the pregnant mouse uterus during late gestation. We observed a marked increase in levels of PR C and transcriptionally active PR-B specifically in fundal myometrium of women in labor. In pregnant mouse uterus, levels of PR-B and PR-C also increased between 15 days post coitum and term, whereas expression of PR-A was dramatically up regulated at 19 days post coitum. In studies of uterine tissues of mice injected intraamniotically with surfactant protein A and of human myometrial and T47D breast cancer cells in culture, up-regulation of PR isoform expression was observed in response to activation of the NF-kappaB pathway. Chromatin immunoprecipitation analysis revealed IL-1beta induced binding of NF-kappaB to the PR promoter. Collectively, these findings suggest that up-regulation of inhibitory PR isoform expression by NF-kappaB activation in both laboring human fundus and pregnant mouse uterus near term may inhibit PR transactivation and thereby lead to a loss of uterine quiescence and the onset of labor. PMID- 16339280 TI - Multiple Alignment of protein structures and sequences for VMD. AB - Multiple Alignment is a new interface for performing and analyzing multiple protein structure alignments. It enables viewing levels of sequence and structure similarity on the aligned structures and performing a variety of evolutionary and bioinformatic tasks, including the construction of structure-based phylogenetic trees and minimal basis sets of structures that best represent the topology of the phylogenetic tree. It is implemented as a plugin for VMD (Visual Molecular Dynamics), which is distributed by the NIH Resource for Macromolecular Modeling and Bioinformatics at the University of Illinois. PMID- 16339281 TI - Survival prediction of diffuse large-B-cell lymphoma based on both clinical and gene expression information. AB - MOTIVATION: It is important to predict the outcome of patients with diffuse large B-cell lymphoma after chemotherapy, since the survival rate after treatment of this common lymphoma disease is <50%. Both clinically based outcome predictors and the gene expression-based molecular factors have been proposed independently in disease prognosis. However combining the high-dimensional genomic data and the clinically relevant information to predict disease outcome is challenging. RESULTS: We describe an integrated clinicogenomic modeling approach that combines gene expression profiles and the clinically based International Prognostic Index (IPI) for personalized prediction in disease outcome. Dimension reduction methods are proposed to produce linear combinations of gene expressions, while taking into account clinical IPI information. The extracted summary measures capture all the regression information of the censored survival phenotype given both genomic and clinical data, and are employed as covariates in the subsequent survival model formulation. A case study of diffuse large-B-cell lymphoma data, as well as Monte Carlo simulations, both demonstrate that the proposed integrative modeling improves the prediction accuracy, delivering predictions more accurate than those achieved by using either clinical data or molecular predictors alone. PMID- 16339282 TI - Histone acetylation and transcriptional regulation in the genome of Saccharomyces cerevisiae. AB - MOTIVATION: In eukaryotic genomes, histone acetylation and thereafter departure from the chromatin are essential for gene transcription initiation. Because gene transcription is tightly regulated by transcription factors, there are some speculations on the cooperation of histone acetylation and transcription factor binding. However, systematic statistical analyses of this relationship on a genomic scale have not been reported. RESULTS: We apply several statistical methods to explore this relationship on two recent genomic datasets: acetylation levels on 11 histone lysines and binding activities of 203 transcription factors, both in promoter regions across the yeast genome. By canonical correlation analysis, we find that a histone acetylation pattern is correlated with a certain profile of transcription factor binding in the genome. Furthermore, after clustering the genes by their acetylation levels on the 11 histone lysines, the genes within clusters show distinct transcription factor binding profiles, as discovered by principle component analysis. Even after applying fairly stringent statistical measurement, most of these clusters have transcription factors with binding activities significantly deviated from the overall genome. We conclude that in the yeast genome, there is a strong correlation between histone acetylation and transcription factor binding in the promoter regions. PMID- 16339283 TI - SubLoc: a server/client suite for protein subcellular location based on SOAP. AB - Based on SOAP(Simple Object Access Protocol) technology, the SubLoc server/client suite offers a user-friendly interface for searching and predicting protein subcellular location. PMID- 16339284 TI - VIS-O-BAC: exploratory visualization of functional genome studies from bacteria. AB - The visualization-aided exploration of complex datasets will allow the research community to formulate novel functional hypotheses leading to a better understanding of biological processes at all levels. Therefore, we have developed a web resource termed VIS-O-BAC designed for the functional investigation of expression data for model systems, such as bacterial pathogens based on a graphical display. Genome-scale datasets derived from typical 'omic' approaches can directly be explored with respect to three biologically relevant aspects, the genome structure (operon organization), the organization of genes in pathways (KEGG) and the gene function with Gene Ontology (GO) terms. The integrated viewers can be used in parallel and combine expression data and functional annotations from different external data repositories. The graphical visualizations evidently accelerate both the validation of regulatory information and the detection of affected biological processes. AVAILABILITY: http://leger2.gbf.de/cgi-bin/vis-o-bac.pl. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 16339285 TI - An extensible automated protein annotation tool: standardizing input and output using validated XML. AB - MOTIVATION: There is a frequent need to apply a large range of local or remote prediction and annotation tools to one or more sequences. We have created a tool able to dispatch one or more sequences to assorted services by defining a consistent XML format for data and annotations. RESULTS: By analyzing annotation tools, we have determined that annotations can be described using one or more of the six forms of data: numeric or textual annotation of residues, domains (residue ranges) or whole sequences. With this in mind, XML DTDs have been designed to store the input and output of any server. Plug-in wrappers to a number of services have been written which are called from a master script. The resulting APATML is then formatted for display in HTML. Alternatively further tools may be written to perform post-analysis. PMID- 16339286 TI - Gene sequence signatures revealed by mining the UniGene affiliation network. AB - BACKGROUND: In the post-genomic era, developing tools to decode biological information from genomic sequences is important. Inspired by affiliation network theory, we investigated gene sequences of two kinds of UniGene clusters (UCs): narrowly expressed transcripts (NETs), whose expression is confined to a few tissues; and prevalently expressed transcripts (PETs) that are expressed in many tissues. RESULTS: We explored the human and the mouse UniGene databases to compare NETs and PETs from different perspectives. We found that NETs were associated with smaller cluster size, shorter sequence length, a lower likelihood of having LocusLink annotations, and lower and more sporadic levels of expression. Significantly, the dinucleotide frequencies of NETs are similar to those of intergenic sequences in the genome, and they differ from those of PETs. We used these differences in dinucleotide frequencies to develop a discriminant analysis model to distinguish PETs from intergenic sequences. CONCLUSIONS: Our results show that most NETs resemble intergenic sequences, casting doubts on the quality of such UniGene clusters. However, we also noted that a fraction of NETs resemble PETs in terms of dinucleotide frequencies and other features. Such NETs may have fewer quality problems. This work may be helpful in the studies of non coding RNAs and in the validation of gene sequence databases. PMID- 16339287 TI - A randomised controlled trial of the reciprocating syringe in arthrocentesis. AB - OBJECTIVE: To evaluate the outcomes of arthrocentesis with the new highly controllable, one handed reciprocating procedure syringe compared with a conventional syringe. METHODS: 100 arthrocentesis procedures were randomised between the reciprocating syringe and the conventional syringe. Outcome measures included patient pain, procedure duration, operator satisfaction, synovial fluid volume, cell counts, and complications. RESULTS: 50 arthrocentesis procedures with the conventional syringe resulted in a mean (SD) procedure time of 3.39 (1.88) minutes, a mean VAPS (patient pain) score of 5.35 (3.15), and a mean VASS (operator satisfaction) score of 4.88 (1.92); 30 of the 50 subjects experienced moderate to severe pain (VAPS score 5 or greater) during arthrocentesis. In contrast, the reciprocating syringe resulted in a reduced procedure time of 1.94 (1.14) minutes (p<0.001), a reduced VAPS (patient pain) score of 2.54 (1.60) (p<0.001), and an increased VASS (operator satisfaction) score of 8.91 (0.79) (p<0.001). Only five of the 50 of subjects experienced moderate to severe pain with the reciprocating syringe. Synovial cell counts were similar between the two syringes (p>0.05), but there was a trend toward greater volume (greater synovial fluid yield) and fewer red blood cells with the reciprocating syringe. CONCLUSIONS: Arthrocentesis with a conventional syringe results in moderate to severe pain in 60% of subjects. The reciprocating syringe prevents significant pain, reduces procedure time, and improves physician performance of arthrocentesis. The reciprocating syringe is superior to the conventional syringe in arthrocentesis. PMID- 16339288 TI - Evidence for differential acquired drug resistance to anti-tumour necrosis factor agents in rheumatoid arthritis. AB - BACKGROUND: Acquired drug resistance or gradual drug failure has been described with most disease modifying antirheumatic drugs (DMARDs) and is also starting to be recognised with anti-tumour necrosis factor (anti-TNF) agents. OBJECTIVE: To study acquired drug resistance to anti-TNF agents in rheumatoid arthritis (RA). METHODS: Swiss health authorities requested continuous monitoring of patients receiving biological agents. Intensification of co-therapy with traditional DMARDs, gradual dose escalation, and drug discontinuation rates in all patients receiving infliximab, etanercept, or adalimumab, adjusting for potential confounders, were analysed. Intensification of DMARD co-therapy and time to discontinuation of the three anti-TNF agents were analysed using a proportional hazards models. Dose escalation and evolution of RA disease activity (DAS28) were analysed using a longitudinal regression model. RESULTS: 1198 patients contributing 1450 patient-years of anti-TNF treatment met the inclusion criteria. The rate of intensification of traditional DMARD co-therapy over time was significantly higher with infliximab (hazards ratio = 1.73 (99% confidence interval (CI) 1.19 to 2.51)) than with the two other agents. Infliximab also showed significant dose escalation over time, with an average dose increase of +12% (99% CI 8% to 16%) after 1 year, and +18% (99% CI 11% to 25%) after 2 years. No significant differences in discontinuation rates were seen between the three anti-TNF agents (ANOVA, p = 0.67). Evolution of disease activity over time indicated a lower therapeutic response to infliximab (DAS28, p<0.001) compared with etanercept, after 6 months' treatment. CONCLUSIONS: In this population, infliximab was associated with a higher risk of requiring intensification of DMARD co-therapy than the other anti-TNF agents and a significant dose escalation over time. Analysis of RA disease activity indicated a reduced therapeutic response to infliximab after the first 6 months of treatment, suggestive of acquired drug resistance. PMID- 16339289 TI - Efficacy and tolerability of once-monthly oral ibandronate in postmenopausal osteoporosis: 2 year results from the MOBILE study. AB - BACKGROUND: Reducing bisphosphonate dosing frequency may improve suboptimal adherence to treatment and therefore therapeutic outcomes in postmenopausal osteoporosis. Once-monthly oral ibandronate has been developed to overcome this problem. OBJECTIVE: To confirm the 1 year results and provide more extensive safety and tolerability information for once-monthly dosing by a 2 year analysis. METHODS: MOBILE, a randomised, phase III, non-inferiority study, compared the efficacy and safety of once-monthly ibandronate with daily ibandronate, which has previously been shown to reduce vertebral fracture risk in comparison with placebo. RESULTS: 1609 postmenopausal women were randomised. Substantial increases in lumbar spine bone mineral density (BMD) were seen in all treatment arms: 5.0%, 5.3%, 5.6%, and 6.6% in the daily and once-monthly groups (50 + 50 mg, 100 mg, and 150 mg), respectively. It was confirmed that all once-monthly regimens were at least as effective as daily treatment, and in addition, 150 mg was found to be better (p<0.001). Substantial increases in proximal femur (total hip, femoral neck, trochanter) BMD were seen; 150 mg produced the most pronounced effect (p<0.05 versus daily treatment). Independent of the regimen, most participants (70.5-93.5%) achieved increases above baseline in lumbar spine or total hip BMD, or both. Pronounced decreases in the biochemical marker of bone resorption, sCTX, observed in all arms after 3 months, were maintained throughout. The 150 mg regimen consistently produced greater increases in BMD and sCTX suppression than the 100 mg and daily regimens. Ibandronate was well tolerated, with a similar incidence of adverse events across groups. CONCLUSIONS: Once-monthly oral ibandronate is at least as effective and well tolerated as daily treatment. Once-monthly administration may be more convenient for patients and improve therapeutic adherence, thereby optimising outcomes. PMID- 16339290 TI - Declines in number of tender and swollen joints in patients with rheumatoid arthritis seen in standard care in 1985 versus 2001: possible considerations for revision of inclusion criteria for clinical trials. AB - OBJECTIVE: To analyse tender and swollen joint counts in three cohorts of patients with rheumatoid arthritis (RA), with a focus on the proportions of patients who had fewer than 6-12 tender or swollen joints, as possible evidence based information toward more generalisable inclusion criteria for current and future RA clinical trials. METHODS: Tender and swollen joint counts were analysed in three cohorts of patients with RA: 125 in 1985, 138 in 2000, and 232 with early RA in 2001. RESULTS: The median numbers of tender joints were 11, 2, and 4 in 1985, 2000, and in early RA in 2001, respectively. The median numbers of swollen joints were 12, 6, and 5 in 1985, 2000, and 2001, respectively. The numbers of tender joints among 28 assessed were >or=12, >or=6, >or=4, and >or=3 in 47%, 80%, 85%, and 90% of patients in 1985; 20%, 37%, 44%, and 49% in 2000; and 17%, 37%, 50%, and 58% in early RA in 2001. The numbers of swollen joints among 28 assessed were >or=12, >or=6, >or=4, and >or=3 in 51%, 78%, 86%, and 90% of patients in 1985; 20%, 50%, 64%, and 67% in 2000; and 14%, 46%, 58%, and 72% in 2001. The number of patients with >or=6 tender or swollen joints in 1985 was greater than the number with >or=3 joints in 2000 and in early RA in 2001. CONCLUSION: Contemporary cohorts of patients seen in standard care have smaller numbers of tender and swollen joints than in previous times. These findings suggest that revision of inclusion criteria for numbers of tender and swollen joints in contemporary RA clinical trials might improve generalisability. PMID- 16339291 TI - Baseline comorbidity levels in biologic and standard DMARD treated patients with rheumatoid arthritis: results from a national patient register. AB - OBJECTIVE: To describe the occurrence of baseline comorbidity in subjects with active rheumatoid arthritis starting treatment with biological agents. Such data are necessary to interpret the reported occurrence of adverse events following treatment. METHODS: Baseline comorbidity was recorded in a large national cohort of patients with rheumatoid arthritis newly starting biological agents. The distribution of the number and types of comorbidities is presented. RESULTS: In all, 7818 patients treated with biological agents (infliximab 3332, etanercept 3302, adalimumab 1059, anakinra 132) were included in the analysis. Comorbidity was common, with 58% of patients having at least one comorbid condition and 25% having more than one. The most frequent comorbid conditions were hypertension, depression, peptic ulcer disease, and respiratory disease. CONCLUSIONS: In routine use, patients treated with biological agents have high levels of baseline comorbidity, which should influence the interpretation of reported adverse events. PMID- 16339292 TI - Urinary levels of type II collagen C-telopeptide crosslink are unrelated to joint space narrowing in patients with knee osteoarthritis. AB - OBJECTIVE: To determine whether urinary concentrations of the cross linked C telopeptide of type II collagen (CTx-II) distinguish subjects with progressive radiographic or symptomatic knee osteoarthritis from those with stable disease. METHODS: SUBJECTS: were 120 obese women with unilateral knee osteoarthritis who participated in a 30 month, randomised, placebo controlled trial of structure modification by doxycycline, in which a standardised semiflexed anteroposterior view of the knee was obtained at baseline and 30 months. Subjects were selected from a larger sample to permit comparisons of urinary CTx-II levels between 60 progressors and 60 non-progressors with respect to medial joint space narrowing. Each group contained 30 subjects who, across five semi-annual assessments, reported on at least two occasions an increase of > or =20% in 50 ft walk pain (minimum = 1 cm on a 10 cm visual analogue scale), relative to the previous visit. The remainder reported no increases in knee pain. Urine samples were obtained semi-annually for determination of the CTx-II and creatinine concentrations. RESULTS: In an analysis of the placebo group only, the frequency of radiographic progressors in the upper and middle tertiles (48% and 60%, respectively) of the baseline CTx-II distribution was not significantly different than that in the lower tertile (64%). These results were unchanged after inclusion of data from subjects in the doxycycline group. Furthermore, serial CTx II levels did not distinguish subjects with progressive radiographic or symptomatic knee osteoarthritis from those with stable disease. CONCLUSIONS: In this pilot study, urinary CTx-II concentration was not a useful biomarker of osteoarthritis progression. PMID- 16339293 TI - Pharmacoeconomic study of patients with chronic inflammatory joint disease before and during infliximab treatment. AB - OBJECTIVE: To evaluate medical and work disability costs for patients with chronic inflammatory joint disease during one year before and one year after institution of infliximab treatment in routine clinical practice. METHODS: Starting from 1999, clinical and laboratory variables for patients treated with biological agents for inflammatory rheumatic diseases were systematically recorded at Helsinki University Central Hospital. From this database clinical information was collected on 96 patients in whom infliximab was started during the period 1999 to 2001. Economic analyses were based on costs incurred because of outpatient and inpatient visits, orthopaedic operations, drugs used, and days on sickness or rehabilitation allowance. Medical and work disability costs were calculated separately for the one year period before (period I) and the one year period after institution of infliximab (period II). RESULTS: Of the study group of 96 patients (arthritis duration 16 years (range 3 to 43)), 74 completed one year of infliximab treatment. Their clinical and laboratory variables improved significantly. The mean increase in medical costs during period II was euro12 015 (95% confidence interval, 6496 to 18,076). A minimal decrease in work disability costs occurred-mean decrease euro130 (-1268 to 1072). CONCLUSIONS: One year treatment with infliximab in patients with longstanding aggressive arthritis showed a good clinical effect but raised medical costs significantly. Work disability costs failed to show a substantial decrease. Starting infliximab in the earlier stages of chronic arthritis could in the long term prevent work disability and thus decrease the total cost to society. PMID- 16339294 TI - Anatomic modifications in the enteric nervous system of piebald mice and physiological consequences to colonic motor activity. AB - It has been assumed that in piebald lethal mice that develop megacolon, impaired colonic motor activity is restricted to the aganglionic distal colon. Peristaltic mechanical recordings, immunohistochemistry, and quantitative PCR were used to investigate whether regions of the colon, other than the aganglionic segment, may also show anatomical modifications and dysfunctional colonic motor activity. Contrary to expectations, colonic migrating motor complexes (MMCs) were absent along the whole colon of piebald lethal homozygote mice and severely impaired in heterozygote siblings. Aganglionosis was detected not only in the distal colon of piebald homozygote lethal mice (mean length: 20.4 +/- 2.1 mm) but also surprisingly in their heterozygote siblings (mean length: 12.4 +/- 1.1 mm). Unlike homozygote lethal mice, piebald heterozygotes showed no signs of megacolon. Interestingly, mRNA expression for PGP 9.5 was also dramatically reduced (by 71-99%) throughout the entire small and large bowel in both homozygote lethal and heterozygous littermates (by 67-87%). Histochemical staining confirmed a significant reduction in myenteric ganglia along the whole colon. In summary, the piebald mutation in homozygote lethal and heterozygote siblings is associated with dramatic reductions in myenteric ganglia throughout the entire colon and not limited to the distal colon as originally thought. Functionally, this results in an absence or severe impairment of colonic MMC activity in both piebald homozygote lethal and heterozygote siblings, respectively. The observation that piebald heterozygotes have an aganglionic distal colon (mean length: 12 mm) but live a normal murine life span without megacolon suggests that aganglionosis >12 mm and the complete absence of colonic MMCs may be required before any symptoms of megacolon arise. PMID- 16339295 TI - Gain of allelic gene expression for IGF-II occurs frequently in Barrett's esophagus. AB - The IGF-II gene normally exhibits genomic imprinting, a DNA modification that allows the expression of only one of the two inherited alleles. With loss of imprinting, there is a gain of allelic gene expression (GOAGE) due to IGF-II being expressed by both alleles. GOAGE for IGF-II has been demonstrated in a number of malignancies and in normal epithelia surrounding malignancies, but not in epithelia without associated neoplasia. We hypothesized that nonneoplastic Barrett's epithelium might have GOAGE for IGF-II that could facilitate its progression to neoplasia. Endoscopic biopsies were obtained from metaplastic esophageal, normal gastric, and normal duodenal epithelia from 43 patients with Barrett's esophagus. Genomic DNA were analyzed using PCR followed by ApaI restriction enzyme digestion or allele-specific PCR to identify an ApaI polymorphism of IGF-II. cDNA from patients with the ApaI polymorphism were analyzed for IGF-II GOAGE using exon connection PCR, followed by a secondary nested PCR and ApaI restriction enzyme digestion. We found that 13 (30%) of 43 samples of Barrett's metaplasia contained the ApaI polymorphism and were thus informative for IGF-II, and sufficient material was available for GOAGE analysis in 9 of those 13 cases. GOAGE for IGF-II was demonstrated in five (56%) of those nine cases. All patients with GOAGE in Barrett's metaplasia also demonstrated GOAGE in the gastric and duodenal epithelia. In contrast, patients without GOAGE in Barrett's metaplasia also had no GOAGE in their gastric and duodenal epithelia. We conclude that in patients with Barrett's esophagus, GOAGE for IGF II is found frequently in the metaplastic esophageal epithelium as well as in normal gastric and duodenal epithelia. PMID- 16339296 TI - Zymogen activation in a reconstituted pancreatic acinar cell system. AB - Pathological activation of digestive zymogens within the pancreatic acinar cell initiates acute pancreatitis. Cytosolic events regulate this activation within intracellular compartments of unclear identity. In an in vivo model of acute pancreatitis, zymogen activation was detected in both zymogen granule-enriched and microsomal cellular fractions. To examine the mechanism of this activation in vitro, a reconstituted system was developed using pancreatic cytosol, a zymogen granule-enriched fraction, and a microsomal fraction. Addition of cytosol to either particulate fraction resulted in a prominent increase in both trypsin and chymotrypsin activities. The percentage of the pool of trypsinogen and chymotrypsinogen activated was about twofold and sixfold greater, respectively, in the microsomal than in the zymogen granule-enriched fraction. Activation of chymotrypsinogen but not trypsinogen was significantly enhanced by ATP (5 mM) but not by the inactive ATP analog AMP-PNP. The processing of procarboxypeptidase B to its mature form also demonstrated a requirement for ATP and cytosol. E64d, an inhibitor of cathepsin B, a thiol protease that can activate trypsin, completely inhibited trypsin activity but did not affect chymotrypsin activity or carboxypeptidase B generation. These studies demonstrate that both zymogen granule-enriched and microsomal fractions from the pancreas can support cytosol dependent zymogen activation. A component of the activation of some zymogens, such as chymotrypsinogen and procarboxypeptidase, may depend on ATP but not on trypsin or cathepsin B. PMID- 16339298 TI - NAD(P)H oxidase contributes to the progression of remote hepatic parenchymal injury and endothelial dysfunction, but not microvascular perfusion deficits. AB - Oxidative stress occurs in remote liver injury, but the origin of the oxidant generation has yet to be thoroughly delineated. Some reports suggest that the source of the distant oxidative stress originates from the site of initial insult [i.e., xanthine oxidase (XO)]; however, it could also be derived from sources such as phagocytic and/or vascular NAD(P)H oxidase (Nox) enzymes. With a murine model of bilateral hindlimb ischemia-reperfusion, we describe here a mechanism for Nox-dependent oxidant production that contributes, at least in part, to remote hepatic parenchymal injury and sinusoidal endothelial cell (SEC) dysfunction. To determine whether Nox enzymes were the source of oxidants, mice were treated immediately after the onset of hindlimb ischemia with specific inhibitors to XO (50 mg/kg ip allopurinol) or Nox (10 mg/kg ip gp91ds-tat and 3 mg/kg ip apocynin). After 1 h of ischemia, hindlimbs were reperfused for either 3 or 6 h. Inhibition of XO failed to provide any improvement in parenchymal injury, SEC dysfunction, neutrophil accumulation, or microvascular dysfunction. In contrast, the inhibition of Nox enzymes prevented the progression (6 h) of parenchymal injury, significantly protected against SEC dysfunction, and completely prevented signs of neutrophil-derived oxidant stress. At the same time, however, inhibition of Nox failed to protect against the early parenchymal injury and microvascular dysfunction at 3 h of reperfusion. These data confirm that microvascular perfusion deficits are not essential for the pathogenesis of remote hepatic parenchymal injury. The data also suggest that Nox enzymes, not XO, are involved in the progression of compromised hepatic parenchymal and endothelial integrity during a systemic inflammatory response. PMID- 16339297 TI - Adrenergic receptor agonists prevent bile duct injury induced by adrenergic denervation by increased cAMP levels and activation of Akt. AB - Loss of parasympathetic innervation after vagotomy impairs cholangiocyte proliferation, which is associated with depressed cAMP levels, impaired ductal secretion, and enhanced apoptosis. Agonists that elevate cAMP levels prevent cholangiocyte apoptosis and restore cholangiocyte proliferation and ductal secretion. No information exists regarding the role of adrenergic innervation in the regulation of cholangiocyte function. In the present studies, we investigated the role of adrenergic innervation on cholangiocyte proliferative and secretory responses to bile duct ligation (BDL). Adrenergic denervation by treatment with 6 hydroxydopamine (6-OHDA) during BDL decreased cholangiocyte proliferation and secretin-stimulated ductal secretion with concomitant increased apoptosis, which was associated with depressed cholangiocyte cAMP levels. Chronic administration of forskolin (an adenylyl cyclase activator) or beta(1)- and beta(2)-adrenergic receptor agonists (clenbuterol or dobutamine) prevented the decrease in cholangiocyte cAMP levels, maintained cholangiocyte secretory and proliferative activities, and decreased cholangiocyte apoptosis resulting from adrenergic denervation. This was associated with enhanced phosphorylation of Akt. The protective effects of clenbuterol, dobutamine, and forskolin on 6-OHDA-induced changes in cholangiocyte apoptosis and proliferation were partially blocked by chronic in vivo administration of wortmannin. In conclusion, we propose that adrenergic innervation plays a role in the regulation of biliary mass and cholangiocyte functions during BDL by modulating intracellular cAMP levels. PMID- 16339299 TI - Febrile-range temperature but not heat shock augments the acute phase response to interleukin-6 in human hepatoma cells. AB - The relationship between the stress protein response and the acute phase response (APPR) was studied in human hepatoma cells to investigate the hierarchy of regulation of these survival responses. Huh-7 cells were subjected to heat treatment (febrile-range temperature 40 degrees C or heat shock 43 degrees C) followed by recovery at 37 degrees C in the presence or absence of IL-6 given either before or after heat treatment. The effects on total, fractional, and acute phase protein synthesis were then analyzed by metabolic labeling, ELISA, real-time PCR, Northern blot analysis, and activation of an alpha(1)-antitrypsin reporter plasmid. Cell energetics were studied under the same conditions using an index of mitochondrial activity and measurement of cellular ATP levels. Febrile range temperature (40 degrees C) augmented acute phase protein production when cells had been pretreated with IL-6. Pretreatment of cells with IL-6 also prevented heat shock-induced suppression of alpha(1)-antichymotrypsin (ACT) but not transferrin. mRNA expression of ACT and alpha(1)-antitrypsin reporter activation studies was consistent with transcriptional regulation of these proteins. Expression of mRNA transcripts for transferrin was increased despite protein expression being reduced by heat shock. The effects of heat shock on acute phase protein synthesis can be modified by preincubation with IL-6, whereas addition of this ligand after heat treatment has no effect on the suppressive effect of heat on the APPR. The mechanism of this action appears to be transcriptionally regulated in the case of ACT, but in the case of transferrin, it may be mediated by another process such as posttranslational modification. PMID- 16339300 TI - Assessment of vitamin D status in male osteoporosis. AB - BACKGROUND: Clinical assessment of vitamin D status often relies on measuring total circulating 25-hydroxyvitamin D3 (25OHD3), but much of each vitamin D metabolite is bound to plasma vitamin D-binding protein (DBP), such that the percentage of free vitamin is very low. We hypothesized that measurement of free rather than total 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and 25OHD3 may provide better assessment of vitamin D status. We therefore aimed to assess vitamin D status in men with idiopathic osteoporosis, in whom possible secondary causes of osteoporosis had been excluded, and to determine the extent of change in biologically active "free" vitamin D caused by variation in plasma DBP concentrations. METHODS: We measured 1,25(OH)2D3 and 25OHD3 in plasma samples from 56 men with idiopathic osteoporosis [mean (SD) age, 59.6 (13.6) years; range, 21-86 years] and 114 male controls [62.4 (10.4) years; range, 44-82 years]. RESULTS: Mean total plasma 25OHD3 in the 56 men with osteoporosis and the 114 controls was 44.7 (21) and 43.3 (17) nmol/L, respectively; total plasma 1,25(OH)2D3 measured in randomly selected men with osteoporosis (n = 50) and controls (n = 50) was 90 (37) and 103 (39) pmol/L, respectively. Mean plasma DBP was significantly higher (P <0.001) in men with osteoporosis [224 (62) mg/L; n = 56] than in the controls [143 (34) mg/L; n = 114], but calculated free plasma 25OHD3 and 1,25(OH)2D3 were significantly lower in the osteoporotic men than in controls [6.1 (3.1) vs 9.1 (4.4) pmol/L (P <0.00001) and 77 (37) vs 142 (58) fmol/L (P <0.00001), respectively]. CONCLUSIONS: Measurement of total vitamin D metabolites alone, although providing a crude assessment of vitamin D status, may not give an accurate indication of the free (biologically active) form of the vitamin. The ratio of total 25OHD3 and 1,25(OH)2D3 to plasma DBP, rather than total circulating vitamin D metabolites, may provide a more useful index of biological activity. Further studies are required to substantiate this hypothesis. PMID- 16339301 TI - Clinically related protein-peptide interactions monitored in real time on novel peptide chips by surface plasmon resonance imaging. AB - BACKGROUND: Developing rapid, high-throughput assays for detecting and characterizing protein-protein interactions is a great challenge in the postgenomic era. We have developed a new method that allows parallel analysis of multiple analytes in biological fluids and is suitable for biological and medical studies. METHODS: This technology for studying peptide-antibody interactions is based on polypyrrole-peptide chips and surface plasmon resonance imaging (SPRi). We generated a chip bearing a large panel of peptide probes by successive electro directed copolymerizations of pyrrole-peptide conjugates on a gold surface. RESULTS: We provide evidence that (a) the signal produced by antibody binding is highly specific; (b) the detected signal specifically reflects the antibody concentration of the tested solution in a dose-dependent manner; (c) this technique is appropriate for analyzing complex media such as undiluted sera, a novelty with respect to previous techniques; and (d) correlation between classic ELISA results and the SPRi signal is good (P = 0.008). We also validated this system in a medical model by detecting anti-hepatitis C antibodies in patient derived sera. CONCLUSION: Because of its characteristics (easy preparation of the peptide chip; high-throughput, label-free, real-time detection; high specificity; and low background), this technology is suitable for screening biological samples and for large-scale studies. PMID- 16339302 TI - Serologic markers in inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is an enduring disease involving mostly young people, with symptoms of bloody diarrhea and abdominal cramps. Several antibodies have been associated with IBD, the 2 most comprehensively studied being autoantibodies to neutrophils (atypical perinuclear anti-neutrophil cytoplasmic antibodies) and anti-Saccharomyces cerevisiae antibodies. This review focuses on the value of these antibodies for diagnosing IBD, differentiating Crohn disease from ulcerative colitis, indeterminate colitis, monitoring disease, defining clinical phenotypes, predicting response to therapy, and as subclinical markers. Pancreatic antibodies and newly identified anti-microbial antibodies (anti-outer membrane porin C, anti-I2, and anti-flagellin) are also reviewed. PMID- 16339303 TI - Sensitive and inexpensive molecular test for falciparum malaria: detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification. AB - BACKGROUND: Malaria is one of the most important parasitic infections in humans. A sensitive diagnostic test for malaria that could be applied at the community level could be useful in programs to control the disease. The aim of the present work was to develop a simple, inexpensive molecular test for Plasmodium falciparum. METHODS: Blood was collected from controls (n = 100) and from patients diagnosed with falciparum malaria infection (n = 102), who were recruited to the study. Heat-treated blood samples were tested by a loop-mediated isothermal amplification (LAMP) assay for P. falciparum. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. To evaluate the assay, DNA from these samples was purified and tested by PCR. Results from the LAMP and PCR assays were compared. RESULTS: The LAMP assay detected P. falciparum directly from heat-treated blood. The quantitative data from the assay correlated to the parasite counts obtained by blood-film microscopic analyses. When we used the PCR assay as the comparison method, the sensitivity and specificity of the LAMP assay were 95% and 99%, respectively. CONCLUSIONS: Unlike PCR, the LAMP assay does not require purified DNA for efficient DNA amplification, thereby reducing the cost and turnaround time for P. falciparum diagnosis. The assay requires only basic instruments, and assay positivity can be verified by visual inspection. PMID- 16339304 TI - Posttranslational conversion of L-serines to D-alanines is vital for optimal production and activity of the lantibiotic lacticin 3147. AB - As a general rule, ribosomally synthesized polypeptides contain amino acids only in the L-isoform in an order dictated by the coding DNA/RNA. Two of a total of only four examples of L to D conversions in prokaryotic systems occur in posttranslationally modified antimicrobial peptides called lantibiotics. In both examples (lactocin S and lacticin 3147), ribosomally encoded L-serines are enzymatically converted to D-alanines, giving rise to an apparent mistranslation of serine codons to alanine residues. It has been suggested that this conversion results from a two-step reaction initiated by a lantibiotic synthetase converting the gene-encoded L-serine to dehydroalanine (dha). By using lacticin 3147 as a model system, we report the identification of an enzyme, LtnJ, that is responsible for the conversion of dha to D-alanine. Deletion of this enzyme results in the residues remaining as dha intermediates, leading to a dramatic reduction in the antimicrobial activity of the producing strain. The importance of the chirality of the three D-alanines present in lacticin 3147 was confirmed when these residues were systematically substituted by L-alanines. In addition, substitution with L-threonine (ultimately modified to dehydrobutyrine), glycine, or L-valine also resulted in diminished peptide production and/or relative activity, the extent of which depended on the chirality of the newly incorporated amino acid(s). PMID- 16339305 TI - Human cancers overexpress genes that are specific to a variety of normal human tissues. AB - We have analyzed gene expression data from three different kinds of samples: normal human tissues, human cancer cell lines, and leukemic cells from lymphoid and myeloid leukemia pediatric patients. We have searched for genes that are overexpressed in human cancer and also show specific patterns of tissue-dependent expression in normal tissues. Using the expression data of the normal tissues, we identified 4,346 genes with a high variability of expression and clustered these genes according to their relative expression level. Of 91 stable clusters obtained, 24 clusters included genes preferentially expressed either only in hematopoietic tissues or in hematopoietic and one to two other tissues; 28 clusters included genes preferentially expressed in various nonhematopoietic tissues such as neuronal, testis, liver, kidney, muscle, lung, pancreas, and placenta. Analysis of the expression levels of these two groups of genes in the human cancer cell lines and leukemias identified genes that were highly expressed in cancer cells but not in their normal counterparts and, thus, were overexpressed in the cancers. The different cancer cell lines and leukemias varied in the number and identity of these overexpressed genes. The results indicate that many genes that are overexpressed in human cancer cells are specific to a variety of normal tissues, including normal tissues other than those from which the cancer originated. It is suggested that this general property of cancer cells plays a major role in determining the behavior of the cancers, including their metastatic potential. PMID- 16339306 TI - Inhibitory coupling specifically generates emergent gamma oscillations in diverse cell types. AB - Networks of inhibitory neurons regulate synchrony during many physiological and pathological oscillations. To explore how these effects depend on cellular, network, and synaptic factors, we developed and validated a semisynthetic inhibitory network that approximates simultaneous activity in multiple neurons by using consecutive responses from single cells. We recorded from three types of neurons, each of which forms interconnected networks in vivo, but has unique intrinsic properties. In all three cell types, fast inhibitory coupling generated emergent gamma oscillations. By contrast, inhibitory coupling desynchronized slower, spindle-frequency responses specifically in thalamic reticular neurons. The emergent gamma-frequency synchronization was also specific to tonic input and did not occur during responses to phasic inputs. These results illustrate how particular features of inhibitory networks (e.g., cell or input type) contribute to their synchronizing or desynchronizing functions. They also demonstrate phenomena (emergent gamma oscillations) that occur robustly in multiple cell types and may thus be a generic feature of inhibitory networks throughout the brain. PMID- 16339307 TI - The corticotropin-releasing factor receptor-1 pathway mediates the negative affective states of opiate withdrawal. AB - The negative affective symptoms of opiate withdrawal powerfully motivate drug seeking behavior and may trigger relapse to heroin abuse. To date, no medications exist that effectively relieve the negative affective symptoms of opiate withdrawal. The corticotropin-releasing factor (CRF) system has been hypothesized to mediate the motivational effects of drug dependence. The CRF signal is transmitted by two distinct receptors named CRF receptor-1 (CRF1) and CRF2. Here we report that genetic disruption of CRF1 receptor pathways in mice eliminates the negative affective states of opiate withdrawal. In particular, neither CRF1 receptor heterozygous (CRF1+/-) nor homozygous (CRF1-/-) null mutant mice avoided environmental cues repeatedly paired with the early phase of opiate withdrawal. These results were not due to altered associative learning processes because CRF1+/- and CRF1-/- mice displayed reliable, conditioned place aversions to environmental cues paired with the kappa-opioid receptor agonist U-50,488H. We also examined the impact of CRF1 receptor-deficiency upon opiate withdrawal induced dynorphin activity in the nucleus accumbens, a brain molecular mechanism thought to underlie the negative affective states of drug withdrawal. Consistent with the behavioral indices, we found that, during the early phase of opiate withdrawal, neither CRF1+/- nor CRF1-/- showed increased dynorphin mRNA levels in the nucleus accumbens. This study reveals a cardinal role for CRF/CRF1 receptor pathways in the negative affective states of opiate withdrawal and suggests therapeutic strategies for the treatment of opiate addiction. PMID- 16339308 TI - Depletion of minichromosome maintenance protein 5 in the zebrafish retina causes cell-cycle defect and apoptosis. AB - In multicellular organisms, the control of genome duplication and cell division must be tightly coordinated. Essential roles of the minichromosome maintenance (MCM) proteins for genome duplication have been well established. However, no genetic model has been available to address the function of MCM proteins in the context of vertebrate organogenesis. Here, we present positional cloning of a zebrafish mcm5 mutation and characterization of its retina phenotype. In the retina, mcm5 expression correlates closely with the pattern of cell proliferation. By the third day of development, mcm5 is down-regulated in differentiated cells but is maintained in regions containing retinal stem cells. We demonstrate that a gradual depletion of maternally derived MCM5 protein leads to a prolonged S phase, cell-cycle-exit failure, apoptosis, and reduction in cell number in mcm5(m850) mutant embryos. Interestingly, by the third day of development, increased apoptosis is detectable only in the retina, tectum, and hindbrain but not in other late-proliferating tissues, suggesting that different tissues may employ distinct cellular programs in responding to the depletion of MCM5. PMID- 16339309 TI - Structure of peptide sex pheromone receptor PrgX and PrgX/pheromone complexes and regulation of conjugation in Enterococcus faecalis. AB - Many bacterial activities, including expression of virulence factors, horizontal genetic transfer, and production of antibiotics, are controlled by intercellular signaling using small molecules. To date, understanding of the molecular mechanisms of peptide-mediated cell-cell signaling has been limited by a dearth of published information about the molecular structures of the signaling components. Here, we present the molecular structure of PrgX, a DNA- and peptide binding protein that regulates expression of the conjugative transfer genes of the Enterococcus faecalis plasmid pCF10 in response to an intercellular peptide pheromone signal. Comparison of the structures of PrgX and the PrgX/pheromone complex suggests that pheromone binding destabilizes PrgX tetramers, opening a 70 bp pCF10 DNA loop required for conjugation repression. PMID- 16339310 TI - RHL1 is an essential component of the plant DNA topoisomerase VI complex and is required for ploidy-dependent cell growth. AB - How cells achieve their final sizes is a pervasive biological question. One strategy to increase cell size is for the cell to amplify its chromosomal DNA content through endoreduplication cycles. Although endoreduplication is widespread in eukaryotes, we know very little about its molecular mechanisms. Successful progression of the endoreduplication cycle in Arabidopsis requires a plant homologue of archaeal DNA topoisomerase (topo) VI. To further understand how DNA is endoreduplicated and how this process is regulated, we isolated a dwarf Arabidopsis mutant, hyp7 (hypocotyl 7), in which various large cell types that in the wild type normally endoreduplicate multiple times complete only the first two rounds of endoreduplication and stall at 8C. HYP7 encodes the RHL1 (ROOT HAIRLESS 1) protein, and sequence analysis reveals that RHL1 has similarity to the C-terminal domain of mammalian DNA topo IIalpha, another type II topo that shares little sequence homology with topo VI. RHL1 shows DNA binding activity in vitro, and we present both genetic and in vivo evidence that RHL1 forms a multiprotein complex with plant topo VI. We propose that RHL1 plays an essential role in the topo VI complex to modulate its function and that the two distantly related topos, topo II and topo VI, have evolved a common domain that extends their function. Our data suggest that plant topo II and topo VI play distinct but overlapping roles during the mitotic cell cycle and endoreduplication cycle. PMID- 16339311 TI - Social relationships, sleep quality, and interleukin-6 in aging women. AB - This study examined the interplay of social engagement, sleep quality, and plasma levels of interleukin-6 (IL-6) in a sample of aging women (n = 74, aged 61-90, M age = 73.4). Social engagement was assessed by questionnaire, sleep was assessed by using the NightCap in-home sleep monitoring system and the Pittsburgh Sleep Quality Index, and blood samples were obtained for analysis of plasma levels of IL-6. Regarding subjective assessment, poorer sleep (higher scores on the Pittsburgh Sleep Quality Index) was associated with lower positive social relations scores. Multivariate regression analyses showed that lower levels of plasma IL-6 were predicted by greater sleep efficiency (P < 0.001), measured objectively and by more positive social relations (P < 0.05). A significant interaction showed that women with the highest IL-6 levels were those with both poor sleep efficiency and poor social relations (P < 0.05). However, those with low sleep efficiency but compensating good relationships as well as women with poor relationships but compensating high sleep efficiency had IL-6 levels comparable to those with the protective influences of both good social ties and good sleep. PMID- 16339312 TI - Modulation of neuropeptide Y expression in adult mice does not affect feeding. AB - Despite numerous experiments showing that administration of neuropeptide Y (NPY) to rodents stimulates feeding and obesity, whereas acute interference with NPY signaling disrupts feeding and promotes weight loss, NPY-null mice have essentially normal body weight regulation. These conflicting observations suggest that chronic lack of NPY during development may lead to compensatory changes that normalize regulation of food intake and energy expenditure in the absence of NPY. To test this idea, we used gene targeting to introduce a doxycycline (Dox) regulated cassette into the Npy locus, such that NPY would be expressed until the mice were given Dox, which blocks transcription. Compared with wild-type mice, adult mice bearing this construct expressed approximately 4-fold more Npy mRNA, which fell to approximately 20% of control values within 3 days after treatment with Dox. NPY protein also fell approximately 20-fold, but the half-life of approximately 5 days was surprisingly long. The biological effectiveness of these manipulations was demonstrated by showing that overexpression of NPY protected against kainate-induced seizures. Mice chronically overexpressing NPY had normal body weight, and administration of Dox to these mice did not suppress feeding. Furthermore, the refeeding response of these mice after a fast was normal. We conclude that, if there is compensation for changes in NPY levels, then it occurs within the time it takes for Dox treatment to deplete NPY levels. These observations suggest that pharmacological inhibition of NPY signaling is unlikely to have long-lasting effects on body weight. PMID- 16339313 TI - Poxvirus multiprotein entry-fusion complex. AB - Poxviruses have evolved elaborate mechanisms for cell entry, assembly, and exocytosis. Recently, four vaccinia virus membrane proteins, namely A21, A28, H2 and L5, were reported to be necessary for cell entry and virus-induced cell-cell fusion but not for virion morphogenesis or attachment of virus particles to cells. Using immunoaffinity purification followed by mass spectrometry, we now show that these four proteins as well as four additional previously uncharacterized putative membrane proteins (A16, G3, G9, and J5) form a stable complex. These proteins fall into two groups: A21, A28, G3, H2, and L5 have an N terminal transmembrane domain, 0-2 intramolecular disulfide bonds, and no sequence similarity, whereas A16, G9, and J5 have a C-terminal transmembrane domain and 4-10 predicted disulfide bonds and are homologous. Studies with conditional-lethal null mutants indicated that the viral membrane was crucial for assembly of the complex and that the absence of individual polypeptide components profoundly decreased complex formation or stability, suggesting a complicated interaction network. Analysis of purified virions, however, demonstrated that the polypeptides of the complex trafficked independently to the viral membrane even under conditions in which the complex itself could not be isolated. All eight proteins comprising the entry-fusion complex are conserved in all poxviruses, suggesting that they have nonredundant functions and that the basic entry mechanism evolved before the division between vertebrate and invertebrate poxvirus species. PMID- 16339314 TI - Topology of biological networks and reliability of information processing. AB - Survival of living cells and organisms is largely based on highly reliable function of their regulatory networks. However, the elements of biological networks, e.g., regulatory genes in genetic networks or neurons in the nervous system, are far from being reliable dynamical elements. How can networks of unreliable elements perform reliably? We here address this question in networks of autonomous noisy elements with fluctuating timing and study the conditions for an overall system behavior being reproducible in the presence of such noise. We find a clear distinction between reliable and unreliable dynamical attractors. In the reliable case, synchrony is sustained in the network, whereas in the unreliable scenario, fluctuating timing of single elements can gradually desynchronize the system, leading to nonreproducible behavior. The likelihood of reliable dynamical attractors strongly depends on the underlying topology of a network. Comparing with the observed architectures of gene regulation networks, we find that those 3-node subgraphs that allow for reliable dynamics are also those that are more abundant in nature, suggesting that specific topologies of regulatory networks may provide a selective advantage in evolution through their resistance against noise. PMID- 16339316 TI - Auditory-visual fusion in speech perception in children with cochlear implants. AB - Speech, for most of us, is a bimodal percept whenever we both hear the voice and see the lip movements of a speaker. Children who are born deaf never have this bimodal experience. We tested children who had been deaf from birth and who subsequently received cochlear implants for their ability to fuse the auditory information provided by their implants with visual information about lip movements for speech perception. For most of the children with implants (92%), perception was dominated by vision when visual and auditory speech information conflicted. For some, bimodal fusion was strong and consistent, demonstrating a remarkable plasticity in their ability to form auditory-visual associations despite the atypical stimulation provided by implants. The likelihood of consistent auditory-visual fusion declined with age at implant beyond 2.5 years, suggesting a sensitive period for bimodal integration in speech perception. PMID- 16339315 TI - The oncogenic properties of mutant p110alpha and p110beta phosphatidylinositol 3 kinases in human mammary epithelial cells. AB - The PIK3CA gene encoding the p110alpha subunit of Class IA phosphatidylinositol 3 kinases (PI3Ks) is frequently mutated in human tumors. Mutations in the PIK3CB gene encoding p110beta, the only other widely expressed Class IA PI3K, have not been reported. We compared the biochemical activity and transforming potential of mutant forms of p110alpha and p110beta in a human mammary epithelial cell system. The two most common tumor-derived alleles of p110alpha, H1047R and E545K, potently activated PI3K signaling. Human mammary epithelial cells expressing these alleles grew efficiently in soft agar and as orthotopic tumors in nude mice. We also examined a third class of mutations in p110alpha, those in the p85 binding domain. A representative tumor-derived p85-binding-domain mutant R38H showed modestly reduced p85 binding and weakly activated PI3K/Akt signaling. In contrast, a deletion mutant lacking the entire p85-binding domain efficiently activated PI3K signaling. When we constructed in p110beta a mutation homologous to the E545K allele of p110alpha, the resulting p110beta mutant was only weakly activated and allowed minimal soft-agar growth. However, a gene fusion of p110beta with the membrane anchor from c-Src was highly active and transforming in both soft-agar and orthotopic nude mouse assays. Thus, although introduction of activating mutations from p110alpha at the corresponding sites in p110beta failed to render the enzyme oncogenic in human cells, the possibility remains that other mutations might activate the beta isoform. PMID- 16339317 TI - Recombinant human nerve growth factor with a marked activity in vitro and in vivo. AB - Recombinant human nerve growth factor (rhNGF) is regarded as the most promising therapy for neurodegeneration of the central and peripheral nervous systems as well as for several other pathological conditions involving the immune system. However, rhNGF is not commercially available as a drug. In this work, we provide data about the production on a laboratory scale of large amounts of a rhNGF that was shown to possess in vivo biochemical, morphological, and pharmacological effects that are comparable with the murine NGF (mNGF), with no apparent side effects, such as allodynia. Our rhNGF was produced by using conventional recombinant DNA technologies combined with a biotechnological approach for high density culture of mammalian cells, which yielded a production of approximately 21.5 +/- 2.9 mg/liter recombinant protein. The rhNGF-producing cells were thoroughly characterized, and the purified rhNGF was shown to possess a specific activity comparable with that of the 2.5S mNGF by means of biochemical, immunological, and morphological in vitro studies. This work describes the production on a laboratory scale of high levels of a rhNGF with in vitro and, more important, in vivo biological activity equivalent to the native murine protein. PMID- 16339318 TI - The viral polymerase mediates adaptation of an avian influenza virus to a mammalian host. AB - Mammalian influenza viruses are descendants of avian strains that crossed the species barrier and underwent further adaptation. Since 1997 in southeast Asia, H5N1 highly pathogenic avian influenza viruses have been causing severe, even fatal disease in humans. Although no lineages of this subtype have been established until now, such repeated events may initiate a new pandemic. As a model of species transmission, we used the highly pathogenic avian influenza virus SC35 (H7N7), which is low-pathogenic for mice, and its lethal mouse-adapted descendant SC35M. Specific mutations in SC35M polymerase considerably increase its activity in mammalian cells, correlating with high virulence in mice. Some of these mutations are prevalent in chicken and mammalian isolates, especially in the highly pathogenic H5N1 viruses from southeast Asia. These activity-enhancing mutations of the viral polymerase complex demonstrate convergent evolution in nature and, therefore, may be a prerequisite for adaptation to a new host paving the way for new pandemic viruses. PMID- 16339320 TI - A new field record for bat longevity. AB - A new longevity record has been established for a free-living bat-41 years for a male Brandt's bat, Myotis brandtii. The exceptional longevity of bats generally, and this species in particular, should make bats of special interest for researchers studying mechanisms of slow aging. PMID- 16339321 TI - The naked mole-rat: a new long-living model for human aging research. AB - Tremendous variation in maximum life span among species overshadows modest increases in longevity resulting from experimental manipulation. Few aging studies focus on long-lived mammals even though these species may expose mechanisms involved in resisting aging. Naked mole-rats (NMRs approximately 35 grams) are the longest-living (>28.3 years) rodents known. This review describes their biology and potential use in aging research. Lifestyle features concur with most evolutionary theories with the exception of the disposable soma theory. Indeed, maximum life span is similar in breeders and nonbreeders, and these highly fecund animals reproduce until they die. Shared characteristics with calorie-restricted, methionine-restricted, and dwarf mice models of extended longevity include reduced body temperature; reduced thyroid, and blood glucose concentrations; and low glycated hemoglobin; in addition to reduced incidence of cancer. Young naked mole-rats surprisingly have high levels of accrued oxidative damage. With their similar longevity quotient to humans, these rodents may provide a novel opportunity to examine mechanisms modulating aging. PMID- 16339322 TI - Oxidative aging and insulin receptor signaling. AB - The life span of nematodes, fruit flies, and mice can be significantly increased (and aging-related changes decreased) by mutations affecting insulin receptor signaling. This effect involves several cellular functions which are negatively regulated by the insulin receptor and thus typically expressed under fasting conditions. This involvement raises the question of whether the insulin independent basal receptor kinase activity in the postabsorptive state can be decreased without compromising the physiologically important response to insulin in the postprandial state. Recent studies have shown that (a) the basal human insulin receptor kinase activity is increased under oxidative conditions in the absence of insulin and (b) insulin signaling in the fasted state can be decreased by cysteine supplementation. Cysteine supplementation has also been shown to improve certain aging-related parameters, suggesting that the average dietary cysteine consumption in Western countries may be suboptimal. These findings provide a conceptual framework that extends the "free radical theory of aging." PMID- 16339323 TI - Prevention of accelerated cell aging in Werner syndrome using a p38 mitogen activated protein kinase inhibitor. AB - We investigated the role of p38 mitogen-activated protein kinase (MAPK) signalling in the accelerated aging of Werner Syndrome (WS) fibroblasts by use of SB203580, a cytokine-suppressive anti-inflammatory drug that targets p38 activity. SB203580 treatment reverts the aged morphology of young WS fibroblasts to that seen in young normal fibroblasts. In addition, SB203580 increases the life span and growth rate of WS fibroblasts to within the normal range. In young WS cells, p38 is activated coincident with an up-regulation of p21(WAF1), and a reduction in the levels of both activated p38 and p21(WAF1) are seen following treatment with SB203580. As these effects are not seen in young normal cells, our data suggest that the abbreviated replicative life span of WS cells is due to a stress-induced, p38-mediated growth arrest that is independent of telomere erosion. With some p38 inhibitors already in clinical trials, our data suggest a potential route to drug intervention in a premature aging syndrome. PMID- 16339324 TI - Effects of caloric restriction and growth hormone resistance on the expression level of peroxisome proliferator-activated receptors superfamily in liver of normal and long-lived growth hormone receptor/binding protein knockout mice. AB - Growth hormone receptor/binding protein knockout (GHR-KO) mice live approximately 40% longer than their normal siblings do. These mice have dramatically reduced plasma levels of insulin-like growth factor 1 (IGF1) and enhanced insulin sensitivity. We examined the expression level of peroxisome proliferator activated receptors (PPARs) and retinoid X receptors family genes in the livers of normal and GHR-KO mice fed ad libitum or subjected to long-term 30% caloric restriction (CR). The levels of PPARgamma and PPARalpha messenger RNA and proteins and the levels of retinoid X receptors messenger RNA were elevated in long-lived GHR-KO mice as compared to normal mice with no major effect of CR in either genotype. These findings suggest that enhanced insulin sensitivity of GHR KO mice may be related to the altered actions of PPARs family members in the liver. The results also indicate that CR may increase insulin sensitivity through a different mechanism. PMID- 16339325 TI - Aged rat myocardium exhibits normal adenosine receptor-mediated bradycardia and coronary vasodilation but increased adenosine agonist-mediated cardioprotection. AB - The purpose of this study was to determine whether aged myocardium exhibits decreased responsiveness to adenosine A1 and A(2a) receptor activation. Studies were conducted in adult (4-6 months) and aged (24-26 months) Fischer 344 x Brown Norway hybrid (F344 x BN) rats. Effects of the adenosine A1/A(2a) agonist AMP579 were measured in isolated hearts and in rats submitted to in vivo regional myocardial ischemia. Aged isolated hearts exhibited lower spontaneous heart rates and higher coronary resistance, as well as normal A1- and A(2a)-mediated responses. There was no difference in control infarct size between adult and aged rats; however, AMP579 treatment resulted in a 50% greater infarct size reduction in aged rats (18 +/- 4% of risk area) compared to adult rats (37 +/- 3%). These findings suggest that adenosine A1 and A(2a) receptor-mediated effects are not diminished in normal aged myocardium, and that aged hearts exhibit increased adenosine agonist-induced infarct reduction. PMID- 16339326 TI - Changes in biological markers of health: older Americans in the 1990s. AB - BACKGROUND: Many studies that show improved health in older adults have relied on subjective measures of health. This article assesses changes in the physiological status of older Americans during the 1990s using biological measures of high-risk for morbidity and mortality. METHODS: Changes in the prevalence of clinically defined, high risk for 10 biological markers were assessed in respondents age 65 years and older from National Health and Nutrition Examination Surveys (NHANES) III (1988-1994) and IV (1999-2000). RESULTS: Some changes in prevalence of high risk values of biological markers indicate improved health among older adults in the 1990s: a 6% reduction in the prevalence of high-risk total cholesterol (p <.001) and a 7% reduction in the prevalence of high-risk homocysteine (p <.001). Other changes indicate worsening health: a 9% increase in the prevalence of high risk systolic blood pressure (p <.01), a 10% increase in obesity (p <.001), and an 8% increase in the prevalence of high-risk C-reactive protein (p <.001). These changes remained significant after adjusting for age, sex, and education. Results of logistic regressions indicate that changes in the frequency of medication usage, medication efficacy, prevalence of chronic disease, and diet explained some of these changes. CONCLUSIONS: Changes in the prevalence of high-risk values of biological markers in the 1990s are mixed. Greater use and effectiveness of lipid-lowering medication has contributed to the reduction in percentage of the population with high-risk lipid levels, and folate supplementation accounted for a decline in the percentage with high-risk homocysteine. However, increases in the percentage with high-risk systolic blood pressure occurred despite an increase in the use of antihypertensive medications, in part because of the limited ability of antihypertensive medications to bring blood pressure below high-risk levels. PMID- 16339327 TI - 25(OH)D Serum levels decline with age earlier in women than in men and less efficiently prevent compensatory hyperparathyroidism in older adults. AB - BACKGROUND: Although a host of factors are known to influence 25-hydroxyvitamin D [25(OH)D] serum levels, few studies addressed the distinctive sex-specific influence of aging, and the age-specific relationship of parathyroid hormone (PTH) with 25(OH)D. The aims of this research were to evaluate changes of 25(OH)D and PTH levels with age in a large population-based sample of men and women and to test the hypothesis that 25(OH)D serum concentrations needed to offset age associated hyperparathyroidism are significantly higher in older than in younger persons. METHODS: In 1107 participants of the InCHIANTI (Invecchiare in Chianti, i.e., Aging in the Chianti area) study, we collected information on dietary intake, daylight exposure, and disability, and measured renal function and serum 25(OH)D and PTH. RESULTS: In women, the age-related decline of 25(OH)D was already evident shortly after age 50, whereas in men it started only after age 70 and was substantially less steep. Age, daylight exposure, winter season, and disability were independent predictors of low 25(OH)D levels. For any given level of 25(OH)D, PTH levels were progressively and consistently higher in older than in younger participants. CONCLUSIONS: These findings suggest that the age associated fall of serum 25(OH)D starts earlier in women than in men and that higher levels of 25(OH)D are required in older compared to younger persons to avoid the age-associated compensatory hyperparathyroidism. PMID- 16339328 TI - Trunk muscle composition as a predictor of reduced functional capacity in the health, aging and body composition study: the moderating role of back pain. AB - BACKGROUND: Cross-sectionally, lower trunk muscle attenuation (higher fat infiltration) has been associated with poorer physical function in older adults. We hypothesize that lower trunk muscle attenuation will be associated with lower functional capacity 3 years later and that back pain status will moderate this relationship. METHODS: The study sample consisted of a biracial cohort of well functioning men (739) and women (788) aged 70-79 years from the Pittsburgh site of the Health, Aging and Body Composition (Health ABC) study. Computed tomography was used to measure trunk muscle area and attenuation of the lumbar paraspinal, lateral abdominal, and rectus abdominus muscles at baseline. The Health ABC Physical Performance Battery (usual and narrow walk, chair stands, and standing balance) was used to measure functional capacity at the first and fourth annual clinic visits. RESULTS: Regardless of back pain status, average trunk muscle attenuation (but not muscle area) was positively associated with overall physical performance, particularly balance (p <.01), in a fully adjusted model. The association between trunk attenuation and functional capacity was significantly stronger in participants with at least moderate back pain in the year prior to baseline (p <.05 for interaction; attenuation x back pain). Participants with moderate to extreme back pain had a greater decline in function over time (p <.05). CONCLUSIONS: Older adults with poorer trunk muscle composition (higher fat infiltration) exhibit reduced functional capacity, especially balance, 3 years later. Improving trunk muscle composition may be an important yet overlooked approach to maintain function and potentially reduce balance impairments, particularly in persons with a history of back pain. PMID- 16339329 TI - Effects of progressive resistance training on body composition in frail older adults: results of a randomized, controlled trial. AB - BACKGROUND: Progressive resistance exercise training (PRT) has been shown to increase muscle strength and fat-free mass (FFM) in elderly persons. Limited information is available regarding the effects of PRT on lean and fat mass in frail elderly persons. METHODS: Ninety-one community-dwelling sedentary men and women, 78 years and older with physical frailty (defined using standardized objective criteria) were enrolled in a 9-month trial of exercise training (ET). Physical frailty was defined as having 2 of the 3 following criteria: modified Physical Performance Test score between 18 and 32, peak aerobic power between 10 and 18 ml/kg/min, or self-report of difficulty or assistance with two instrumental activities of daily living or one basic activity of daily living. Participants were randomly assigned to either a control (CTL) group that performed a low intensity home exercise program or a supervised ET group that performed 3 months of low intensity exercise and 3 months of PRT. RESULTS: After completion of PRT, ET participants had greater improvements than did CTL participants in maximal voluntary force production for knee extension (mean Delta +5.3 +/- 13 ft/lb vs +1.1 +/- 11 ft/lb, p =.05), measured using isokinetic dynamometry. Total body FFM (measured using dual energy x-ray absorptiometry) increased in the ET group, but not in the CTL group (mean Delta +0.84 +/- 1.4 kg vs +0.01 +/- 1.5 kg, p =.005). Total, trunk, intra-abdominal, and subcutaneous fat mass (measured using dual energy x-ray absorptiometry and (1)H-magnetic resonance imaging) did not change in response to PRT. CONCLUSIONS: Three months of supervised PRT induced improvements in maximal voluntary thigh muscle strength and whole body FFM in frail, community-dwelling elderly women and men. This supervised exercise program may not be sufficient to reduce whole-body or intra abdominal fat area in this population. PMID- 16339330 TI - Exercise accelerates wound healing among healthy older adults: a preliminary investigation. AB - BACKGROUND: Older adults are likely to experience delayed rates of wound healing, impaired neuroendocrine responsiveness, and increased daily stress. Exercise activity has been shown to have a positive effect on physiological functioning and psychological functioning among older adults. This study evaluated the effect of a 3-month exercise program on wound healing, neuroendocrine function, and perceived stress among healthy older adults. METHODS: Twenty-eight healthy older adults (mean age 61.0 +/- 5.5 years) were assigned randomly to an exercise activity group (n = 13) or to a nonexercise control group (n = 15). One month following baseline randomization, after exercise participants had acclimated to the exercise routine, all participants underwent an experimental wound procedure. Wounds were measured 3 times per week until healed to calculate rate of wound healing. All participants completed assessments of exercise endurance, salivary cortisol, and self-reported stress prior to randomization and at the conclusion of the intervention. RESULTS: Exercise participants achieved significant improvements in cardiorespiratory fitness, as reflected by increased oxygen consumption (VO(2)max) and exercise duration. Wound healing occurred at a significantly faster rate in the exercise group [mean = 29.2 (9.0) days] than in the nonexercise group [38.9 (7.4) days; p =.012]. Exercise participants also experienced increased cortisol secretion during stress testing following the intervention. Group differences in wound healing and neuroendocrine responsiveness were found despite low levels of self-reported stress. CONCLUSIONS: A relatively short-term exercise intervention is associated with enhanced rates of wound healing among healthy older adults. Thus, exercise activity may be an important component of health care to promote wound healing. PMID- 16339331 TI - Angiotensin-converting enzyme inhibition intervention in elderly persons: effects on body composition and physical performance. AB - The disablement process is often accompanied by sarcopenia or muscle loss, which is associated with virtually all identified disability risk factors. Clinically, the association between body composition and physical performance has been documented by several studies. However, loss of strength is greater than loss of muscle mass with age implying that the quality of remaining muscle may be reduced. Although there are limited data explaining potential physiological mechanisms that contribute to muscle quality, sarcopenia is frequently associated with fat accumulation, and the percentage of body fat increases with age even if weight does not. However, the relationship between fat and muscle function may not be linear, suggesting that there may be an optimal ratio of lean to fat mass for physical function. There are no definitive pharmacological interventions proven to prevent decline in physical function either by modulating body composition or by other means. One exception may be angiotensin-converting enzyme inhibitors (ACEIs). ACE is an important component of the renin-angiotensin system, the central hormonal regulator of blood pressure. Recent evidence suggests that ACEIs may improve physical function by means of direct effects on body composition in older persons, rather than through its blood-pressure lowering effects. Clinical and genetic studies in humans and experimental evidence in animals suggest that modulation of the renin-angiotensin system is associated with metabolic and biochemical changes in skeletal muscle and fat, changes that are associated with declining physical function. ACEIs may modulate this process through a variety of molecular mechanisms including their influence on oxidative stress and on metabolic and inflammation pathways. This review describes potential biological mechanisms of ACE inhibition and its contribution to declining physical performance and changing body composition. Promising pharmacoepidemiological studies and experimental evidence in animals suggest that there are appropriate models in which to study this effect. PMID- 16339332 TI - Lower extremity strength plays only a small role in determining the maximum recoverable lean angle in older adults. AB - BACKGROUND: The purpose of this study was to determine the extent to which measures of lower extremity strength and power contribute to the ability of older men and women to restore postural equilibrium using a single-step recovery following a large postural disturbance. METHODS: The postural disturbance, which has been used as a surrogate for forward-directed falls, involved a sudden release from a forward-leaning angle. The ability to recover using a single step was evaluated as the maximum recoverable lean angle for 56 healthy older women and men. Maximum voluntary isometric and isokinetic strength was measured for ankle plantarflexion and dorsiflexion, knee flexion and extension, and hip flexion and extension. Discriminant analysis was used to determine the strength measures that best classified participants as members of the highest (n = 14) or lowest (n = 14) quartiles of maximum recoverable lean angle. Those variables were subsequently entered into a regression analysis to characterize the relationship between strength and maximum recoverable lean angle for the entire participant sample. RESULTS: Maximum isokinetic dorsiflexion strength at 90 degrees /s satisfied the criteria of the stepwise discriminant analysis, and correctly classified 82.1% of the participants in the highest or lowest quartiles of maximum recoverable lean angle. The multiple regression procedure, performed on all participants (n = 56) revealed a significant quadratic relationship between maximum isokinetic dorsiflexion strength at 90 degrees /s and maximum recoverable lean angle (R2 = 0.295; p <.001). CONCLUSIONS: Lower extremity strength makes a small, but significant contribution to maximum recoverable lean angle. However, because 70% of the shared variability remained unaccounted for, it is suggested that other performance factors, such as coordination, may be of greater importance to performance of this time-critical motor task. PMID- 16339333 TI - Adverse events associated with testosterone replacement in middle-aged and older men: a meta-analysis of randomized, placebo-controlled trials. AB - BACKGROUND: We performed a meta-analysis of randomized clinical trials to determine the risks of adverse events associated with testosterone replacement in older men. METHODS: The MEDLINE database was searched from 1966 to April 2004, using testosterone as the indexing term; limits included human, male, > or =45 years old, and randomized controlled trial. Of the 417 studies thus identified, 19 met the inclusion criteria: testosterone replacement for at least 90 days, men > or =45 years old with low or low-normal testosterone level, randomized controlled trial, and medically stable men. Odds ratios (ORs) were pooled using a random effects model, assuming heterogeneous results across studies, and were weighted for sample size. RESULTS: In the 19 studies that met eligibility criteria, 651 men were treated with testosterone and 433 with placebo. The combined rate of all prostate events was significantly greater in testosterone treated men than in placebo-treated men (OR = 1.78, 95% confidence interval [CI], 1.07-2.95). Rates of prostate cancer, prostate-specific antigen (PSA) >4 ng/ml, and prostate biopsies were numerically higher in the testosterone group than in the placebo group, although differences between the groups were not individually statistically significant. Testosterone-treated men were nearly four times as likely to have hematocrit >50% as placebo-treated men (OR = 3.69, 95% CI, 1.82 7.51). The frequency of cardiovascular events, sleep apnea or death was not significantly different between the two groups. CONCLUSIONS: Testosterone replacement in older men was associated with a significantly higher risk of detection of prostate events and of hematocrit >50% than was placebo; hematocrit increase was the most frequent adverse event associated with testosterone replacement. These data reaffirm the need to monitor hematocrit, PSA, and digital examination of the prostate during testosterone replacement in older men. PMID- 16339334 TI - Folate, but not homocysteine, predicts the risk of fracture in elderly persons. AB - BACKGROUND: Recent prospective studies reported that increased plasma homocysteine levels are an independent predictor of osteoporotic fracture in elderly persons. These studies, however, did not take into account folate and vitamin B12, which are the major nutritional determinants of homocysteinemia. METHODS: Incident osteoporotic fractures were assessed in 702 Italian participants aged 65-94 years with a mean follow-up of 4 years (1999/2000 2003/2004). A multivariable logistic regression model was used to study the relation of baseline plasma homocysteine, serum folate, and serum vitamin B12 with risk of fracture. RESULTS: After adjustment for sociodemographic and clinical confounders, the odds ratio (OR) for each increase of 1 standard deviation in log-transformed plasma homocysteine was 1.39 (95% confidence interval [CI], 1.01-1.91), but decreased to 1.22 (95% CI, 0.85-1.74) after further adjustment for serum folate and vitamin B12. The corresponding multivariable-adjusted OR for hyperhomocysteinemia (plasma total homocysteine [tHcy] > 15 micromoL) was 1.58 (95% CI, 0.71-3.53). Participants in the lowest serum folate quartile (< or =9.3 nmol/L) had an increased risk of fracture than did those in higher quartiles (multivariable-adjusted OR = 2.06; 95% CI. 1.02 4.18), but no dose-related protective effect for increasing serum folate levels was found (multivariable-adjusted OR = 0.84 for each increase of 1 standard deviation in log-transformed serum folate, 95% CI, 0.59-1.19). No independent association was found for serum vitamin B12. CONCLUSIONS: Low serum folate is responsible for the association between homocysteine and risk of osteoporotic fracture in elderly persons. PMID- 16339335 TI - The variable component of lateral body sway during walking in young and older humans. AB - INTRODUCTION: Because sideways falls are common in elderly persons, we devised a method to measure variable lateral movements of the thorax with respect to foot position during normal walking. METHODS: Movements of the ankles and shoulders were measured during walking a distance of 9 m. Two age groups were studied: young (n = 17, 6 male, mean age 27.3 years) and older (n = 21, 13 male, mean age 72.7 years) people. During walking, the path followed in the horizontal plane by the midpoint between the two shoulders was compared to the line connecting successive positions of the ankles during stance. Lateral deviations between these two paths were divided into a regular component (average of about 30 strides) and a variable component (the difference between the deviation during each stride and the average). Lateral sway was also observed while standing with eyes open for 1 minute. RESULTS: The older group had more lateral movement during walking in the variable component (p =.006) and a nonsignificant trend (p =.054) in the same direction in the regular component. Eight of the older participants had a value for the variable component greater than the 95% confidence limit for the young participants. Only two of the older participants had a standing sway outside the confidence limit for the young participants. The variable component was associated with variability in stride width. DISCUSSION: The variable component of lateral sway during walking provides good discrimination between age groups, as does variability in step width. It remains to be seen whether these variables are different in fallers and nonfallers. PMID- 16339336 TI - Strategies, time, and costs associated with the recruitment and enrollment of nursing home residents for a micronutrient supplementation clinical trial. AB - BACKGROUND: Concomitant with the substantial growth of the elderly population in the last decade, there has been a steady rise in the number of nursing home residents aged 65 years and older. Well designed, rigorously conducted clinical intervention trials provide an important source of data for evidence-based improvements in the medical care of nursing home residents. The information available on strategies for the recruitment and screening of participants for such studies in long-term care facilities, as well as the financial and time costs for carrying out these investigations, is limited. METHODS: This report describes our experience in recruiting 617 nursing home residents for a multisite, double-blind, randomized, placebo-controlled trial designed to determine the efficacy of a 1-year period of vitamin E supplementation in preventing respiratory tract infections. Comparisons of the projected staffing costs and actual costs incurred are presented, using a retrospective method for the determination of unit costs. RESULTS: Initially, 874 consents were obtained from 2815 potential participants, of which only 617 were enrolled. Each successful enrollment required an average of 15 hours of staff time at a combined personnel and supply cost of $515 per participant and a total study cost of $317,661. Several obstacles were encountered during the recruitment and enrollment process: resistance on the part of family or primary care provider; transfer out of facility; and changes in the medical condition of the patient, including death. DISCUSSION: The results of this report should prove useful to investigators developing budgets for nursing home-based clinical trials by providing a more accurate determination of the personnel needed and the costs associated with recruitment and enrollment of participants. PMID- 16339337 TI - The effect of the presence of others on caloric intake in homebound older adults. AB - BACKGROUND: Undernutrition in homebound older adults is a significant problem. The purpose of this study was to investigate the effect of the presence of others, both within the household and during meals, on caloric intake in homebound older adults. METHODS: In-depth interviews and three 24-hour dietary recalls were obtained from 50 older adults who were receiving home health services. Descriptive statistics were used to characterize participants, and hierarchical linear modeling was performed to evaluate predictors of caloric intake per meal. RESULTS: Participants' mean age was 77. Females composed 65% and African Americans composed 42% of the sample. Analyses are based on 553 meal observations. The majority (84%) of participants consumed all meals for each of the 3 days of data collection; however, they consumed an average of only 1305 calories per day. Hierarchical linear modeling analysis indicated that persons who had others present during meals consumed an average of 114.0 calories more per meal than those who ate alone (p =.009) and that women consumed 76.7 fewer calories per meal than did men (p =.045). The presence of others within the household had no effect on caloric intake. CONCLUSION: This research suggests that a simple and inexpensive way to increase caloric intake in homebound older adults is to make arrangements for family members or caregivers to eat with them. PMID- 16339338 TI - Moderate alcohol consumption is associated with reduced arterial stiffness in older adults: the Rotterdam study. AB - BACKGROUND: Light-to-moderate alcohol consumption has been associated with a lower risk of cardiovascular disease. The protective effect of alcohol could involve arterial properties as arterial stiffness and distensibility. METHODS: The relationship between alcohol and arterial stiffness was studied within the framework of the Rotterdam Study, a population-based study in individuals aged 55 and older. The present study included 3178 participants in the third examination phase. Arterial stiffness was measured by two different methods, i.e., the carotid-femoral pulse wave velocity and the DC of the common carotid artery. Categories of alcohol consumption were defined as follows: < or =3 glasses of alcohol per week, 4-10 glasses per week, 11-20 glasses per week, and > or =21 glasses per week. Linear regression analysis was used to investigate the association between alcohol consumption and measures of arterial stiffness. RESULTS: In multivariate-adjusted models, women drinking 4-10, 11-20, and > or =21 glasses of alcoholic beverage per week had a -0.07 (0.22 to -0.38), -0.18 (0.12 to -0.49), and 0.12 (0.19 to -0.43) m/s difference in mean pulse wave velocity compared to those drinking 0-3 glasses per week (reference group). Corresponding differences in the carotid DC were 0.68 (1.21 to 0.15), 0.28 (0.82 to -0.25), and 0.36 (0.91 to -0.18) 10(-3)/kPa. In men, the estimates were not statistically significant, although a similar trend was observed. CONCLUSIONS: Moderate alcohol consumption is associated with lower arterial stiffness in women independently of cardiovascular risk factors and atherosclerosis. PMID- 16339339 TI - Regional cerebral autoregulation during orthostatic stress: age-related differences. AB - BACKGROUND: We used transcranial Doppler ultrasonography to examine the cerebral blood flow response to orthostatic stress in the middle and posterior cerebral circulations and to determine the effects of healthy aging on regional cerebral blood flow regulation. METHODS: Continuous simultaneous middle (MCA) and posterior (PCA) cerebral artery blood flow velocities (BFV) and mean arterial pressure (MAP) were measured in response to standing from a sitting position in 13 younger (30 +/- 7 years) and 13 older (73 +/- 4 years) healthy participants. RESULTS: The older participants had a significantly larger decline in MAP (-31% +/- 3 in the older and -21% +/- 2 in the younger) and a smaller increase in heart rate (HR) (15 bpm +/- 1 in the older, 24 bpm +/- 2 in the younger) during the posture change. Despite a larger decline in MAP, the older participants had a decline very similar to that of the younger participants in BFVs in both vascular territories. This was associated with a significantly larger vasodilatory response in the MCA and PCA vascular territories of the older participants. There were no regional differences of the cerebrovascular resistance and BFV responses to orthostasis in the younger participants. However, in the older participants, there was a significantly larger BFV decline and a smaller vasodilatory response in the PCA as compared to the MCA territory. CONCLUSIONS: Healthy aging is associated with preserved cerebrovascular adaptation to orthostatic hypotension. However, in older persons, the PCA territory blood flow may be more vulnerable to reduced perfusion during orthostatic stress. PMID- 16339340 TI - Muscle size and strength are increased following walk training with restricted venous blood flow from the leg muscle, Kaatsu-walk training. AB - Previous studies have shown that low-intensity resistance training with restricted muscular venous blood flow (Kaatsu) causes muscle hypertrophy and strength gain. To investigate the effects of daily physical activity combined with Kaatsu, we examined the acute and chronic effects of walk training with and without Kaatsu on MRI-measured muscle size and maximum dynamic (one repetition maximum) and isometric strength, along with blood hormonal parameters. Nine men performed Kaatsu-walk training, and nine men performed walk training alone (control-walk). Training was conducted two times a day, 6 days/wk, for 3 wk using five sets of 2-min bouts (treadmill speed at 50 m/min), with a 1-min rest between bouts. Mean oxygen uptake during Kaatsu-walk and control-walk exercise was 19.5 (SD 3.6) and 17.2 % (SD 3.1) of treadmill-determined maximum oxygen uptake, respectively. Serum growth hormone was elevated (P < 0.01) after acute Kaatsu walk exercise but not in control-walk exercise. MRI-measured thigh muscle cross sectional area and muscle volume increased by 4-7%, and one repetition maximum and maximum isometric strength increased by 8-10% in the Kaatsu-walk group. There was no change in muscle size and dynamic and isometric strength in the control walk group. Indicators of muscle damage (creatine kinase and myoglobin) and resting anabolic hormones did not change in both groups. The results suggest that the combination of leg muscle blood flow restriction with slow-walk training induces muscle hypertrophy and strength gain, despite the minimal level of exercise intensity. Kaatsu-walk training may be a potentially useful method for promoting muscle hypertrophy, covering a wide range of the population, including the frail and elderly. PMID- 16339341 TI - Use of ultrasound to make noninvasive in vivo measurement of continuous changes in human muscle contractile length. AB - Continuous measurement of contractile length has been traditionally achieved using animal preparations in which the muscle and tendon are exposed. More modern methods, e.g., sonomicroscopy, are still invasive. There is a widely perceived need for a noninvasive, in vivo method of measuring continuous changes of human muscle contractile length. Ultrasonography has been used for several years to measure relatively static, discrete changes in tendon, aponeurosis, and muscle fascicle length. We have recently developed this technique to continuously track changes in muscle contractile length during quiet standing. Here, we present the tracking algorithm and use externally applied perturbations to establish the spatial and temporal resolution of the technique. Subjects maintained a low level of ankle torque while a pneumatic actuator applied rapid, square-pulse ankle rotations of defined magnitude and 0.2-s duration. Tracked changes in gastrocnemius and soleus contractile length follow the temporal profile of the perturbations and scale progressively (5-400 microm) with the size of the ankle rotation (0.03-0.7 degrees ). In a second experiment, we tracked a wire oscillating in water with known peak to peak amplitudes of 1.5 microm to 8 mm. The ultrasound tracking procedure had near 100% accuracy at all amplitudes for frequencies up to 3 Hz and showed attenuation at higher frequencies consistent with an effective sampling frequency of 12 Hz and sampling time of 80 ms. This noninvasive technique is sensitive, without averaging, to changes as small as 1 microm and is suitable for observing neuromotor activity in posture and locomotion. PMID- 16339342 TI - Brisk production of nitric oxide and associated formation of S-nitrosothiols in early hemorrhage. AB - The results of previous inhibitor studies suggest that there is some increase in nitric oxide (NO) production from constitutive NO synthase in early hemorrhage (H), but the magnitude of NO production early after H has not been previously assessed. It is generally believed that only modest production rates are possible from the constitutively expressed NO synthases. To study this, anesthetized male Sprague-Dawley rats were subjected to 90 min of isobaric (40 mmHg) H. During this period of time, the dynamics of accumulation of NO intermediates in the arterial blood was assessed using electron paramagnetic resonance spectroscopy, chemiluminescence, fluorescence imaging, and mass spectrometry. Electron paramagnetic resonance-detectable NO adducts were also measured with spin traps in blood plasma and red blood cells. H led to an increase in the concentration of hemoglobin-NO from 0.9 +/- 0.2 to 4.8 +/- 0.7 microM. This accumulation was attenuated by a nonselective inhibitor of NO synthase, NG-nitro-L-argininemethyl ester (L-NAME), but not by NG-nitro-D-argininemethyl ester (D-NAME) or 1400W. Administration of L-NAME (but not 1400W or D-NAME) during H produced a short-term increase in mean arterial pressure ( approximately 90%). In H, the level of N oxides in red blood cells increased sevenfold. S-nitrosylation of plasma proteins was revealed with "biotin switch" techniques. The results provide compelling evidence that there is brisk production of NO in early H. The results indicate that the initial compensatory response to H is more complicated than previously realized, and it involves an orchestrated balance between intense vasoconstrictor and vasodilatory components. PMID- 16339343 TI - Voluntary muscle activation is impaired by core temperature rather than local muscle temperature. AB - Fatigue during hyperthermia may be due in part to a failure of the central nervous system to fully activate the working muscles. We investigated the effects of passive hyperthermia on maximal plantar flexor isometric torque (maximal isometric voluntary contraction) and voluntary activation to determine the roles of local skin temperature, core temperature, and peripheral muscle temperature in fatigue. Nine healthy subjects were passively heated from 37.2 to 39.5 degrees C (core temperature) and then cooled back down to 37.9 degrees C using a liquid conditioning garment, with the right leg kept at a thermoneutral temperature throughout the protocol, whereas the left leg was allowed to heat and cool. Passive heating resulted in significant decreases in torque from [mean (SD)] 172 N x m (SD 39) to 160 N x m (SD 44) and in voluntary activation from 96% (SD 2) to 91% (SD 5) in the heated leg, and maximal isometric voluntary contraction decreased similarly from 178 N xm (SD 37) to 165 N x m (SD 38) and voluntary activation from 97% (SD 2) to 94% (SD 5) in the thermoneutral leg. The initiation of cooling, which produced a rapid decrease in skin temperature and cardiovascular strain [heart rate reserve decreased from 58% (SD 12) to 31% (SD 12)], did not immediately restore either torque or voluntary activation. However, when core temperature was lowered back to normal, torque and voluntary activation were restored to baseline values. It was concluded that an increase in core temperature is a factor responsible for reducing voluntary activation during brief voluntary isometric contractions and that temperature-induced changes in the contractile properties of muscle and local thermal afferent input from the skin do not contribute significantly to the decrement in torque. PMID- 16339344 TI - Precooling leg muscle improves intermittent sprint exercise performance in hot, humid conditions. AB - We used three techniques of precooling to test the hypothesis that heat strain would be alleviated, muscle temperature (Tmu) would be reduced, and as a result there would be delayed decrements in peak power output (PPO) during exercise in hot, humid conditions. Twelve male team-sport players completed four cycling intermittent sprint protocols (CISP). Each CISP consisted of twenty 2-min periods, each including 10 s of passive rest, 5 s of maximal sprint against a resistance of 7.5% body mass, and 105 s of active recovery. The CISP, preceded by 20 min of no cooling (Control), precooling via an ice vest (Vest), cold water immersion (Water), and ice packs covering the upper legs (Packs), was performed in hot, humid conditions (mean +/- SE; 33.7 +/- 0.3 degrees C, 51.6 +/- 2.2% relative humidity) in a randomized order. The rate of heat strain increase during the CISP was faster in Control than Water and Packs (P < 0.01), but it was similar to Vest. Packs and Water blunted the rise of Tmu until minute 16 and for the duration of the CISP (40 min), respectively (P < 0.01). Reductions in PPO occurred from minute 32 onward in Control, and an increase in PPO by approximately 4% due to Packs was observed (main effect; P < 0.05). The method of precooling determined the extent to which heat strain was reduced during intermittent sprint cycling, with leg precooling offering the greater ergogenic effect on PPO than either upper body or whole body cooling. PMID- 16339345 TI - Plasma arginine-vasopressin following experimental stroke: effect of osmotherapy. AB - Neurohumoral responses have been implicated in the pathogenesis of ischemia evoked cerebral edema. In a well-characterized animal model of ischemic stroke, the present study was undertaken to 1) study the profile of plasma arginine vasopressin (AVP), and 2) determine whether osmotherapy with mannitol and various concentrations of hypertonic saline (HS) solutions influence plasma AVP levels. Halothane-anesthetized adult male Wistar rats were subjected to 2 h of middle cerebral artery occlusion with the intraluminal filament technique. Plasma AVP levels (means +/- SD) were significantly elevated at 24 h (42 +/- 21 pg/ml), 48 h (50 +/- 28 pg/ml), and 72 h (110 +/- 47 pg/ml), and returned to baseline at 96 h (22 +/- 15 pg/ml) following middle cerebral artery occlusion compared with sham operated controls (14 +/- 7 pg/ml). Plasma AVP levels at 72 h were significantly attenuated with 7.5% HS (37 +/- 8 pg/ml; 360 +/- 11 osmol/l) compared with 0.9% saline (73 +/- 6; 292 +/- 6 osmol/l), 3% HS (66 +/- 8 pg/ml; 303 +/- 12 osmol/l), or mannitol (74 +/- 9 pg/ml; 313 +/- 14 osmol/l) treatment. HS (7.5%) significantly attenuated water content in the ipsilateral and contralateral hemispheres compared with surgical shams, 0.9% saline, 3% HS, and mannitol treatments. Peak plasma AVP levels were not associated with direct histopathological injury to the anterior hypothalamus. Attenuation of brain water content with 7.5% HS treatment coincides with attenuated serum AVP levels, and we speculate that this may represent one additional mechanism by which osmotherapy attenuates edema associated with ischemic stroke. PMID- 16339346 TI - Acute vibration increases alpha2C-adrenergic smooth muscle constriction and alters thermosensitivity of cutaneous arteries. AB - The vascular symptoms of hand-arm vibration syndrome, including cold-induced vasospasm, are in part mediated by increased sensitivity of cutaneous arteries to sympathetic stimulation. The goal of the present study was to use a rat tail model to analyze the effects of vibration on vascular function and alpha adrenoceptor (AR) responsiveness. Rats were exposed to a single period of vibration (4 h, 125 Hz, constant acceleration 49 m/s2 root mean square). The physical or biodynamic response of the tail demonstrated increased transmissibility or resonance at this frequency, similar to that observed during vibration of human fingers. Morphological analysis demonstrated that vibration did not appear to cause structural injury to vascular cells. In vitro analysis of vascular function demonstrated that constriction to the alpha1-AR agonist phenylephrine was similar in vibrated and control arteries. In contrast, constriction to the alpha2-AR agonist UK14304 was increased in vibrated compared with control arteries, both in endothelium-containing or endothelium-denuded arteries. The alpha2C-AR antagonist MK912 (3 x 10(-10) M) inhibited constriction to UK14304 in vibrated but not control arteries, reversing the vibration-induced increase in alpha2-AR activity. Moderate cooling (to 28 degrees C) increased constriction to the alpha2-AR agonist in control and vibrated arteries, but the magnitude of the amplification was less in vibrated compared with control arteries. Endothelium-dependent relaxation to acetylcholine was similar in control and vibrated arteries. Based on these results, we conclude that a single exposure to vibration caused a persistent increase in alpha2C-AR-mediated vasoconstriction, which may contribute to the pathogenesis of vibration-induced vascular disease. PMID- 16339347 TI - Effects of low-intensity resistance exercise with slow movement and tonic force generation on muscular function in young men. AB - We investigated the acute and long-term effects of low-intensity resistance exercise (knee extension) with slow movement and tonic force generation on muscular size and strength. This type of exercise was expected to enhance the intramuscular hypoxic environment that might be a factor for muscular hypertrophy. Twenty-four healthy young men without experience of regular exercise training were assigned into three groups (n = 8 for each) and performed the following resistance exercise regimens: low-intensity [ approximately 50% of one repetition maximum (1RM)] with slow movement and tonic force generation (3 s for eccentric and concentric actions, 1-s pause, and no relaxing phase; LST); high intensity ( approximately 80% 1RM) with normal speed (1 s for concentric and eccentric actions, 1 s for relaxing; HN); low-intensity with normal speed (same intensity as for LST and same speed as for HN; LN). In LST and HN, the mean repetition maximum was 8RM. In LN, both intensity and amount of work were matched with those for LST. Each exercise session consisting of three sets was performed three times a week for 12 wk. In LST and HN, exercise training caused significant (P < 0.05) increases in cross-sectional area determined with MRI and isometric strength (maximal voluntary contraction) of the knee extensors, whereas no significant changes were seen in LN. Electromyographic and near-infrared spectroscopic analyses showed that one bout of LST causes sustained muscular activity and the largest muscle deoxygenation among the three types of exercise. The results suggest that intramuscular oxygen environment is important for exercise-induced muscular hypertrophy. PMID- 16339348 TI - Direct vasoactive and vasoprotective properties of anthocyanin-rich extracts. AB - Reactive oxygen species (ROS) play a critical role in the impairment of nitric oxide-mediated vascular functions and overall pathogenesis associated with cardiovascular disease. Plant pigment anthocyanins are exceptionally potent oxygen radical scavengers that produce beneficial effects in diseases outside the cardiovascular system. We examined for the first time the potential coronary vasoactive and vasoprotective properties of three anthocyanin enhanced extracts prepared from chokeberry (Ck), bilberry (B), or elderberry (E). Coronary arterial rings were isolated from 64 pigs and incubated in sterile tissue culture media overnight for use in one of four separate in vitro isometric force recording studies. Ck and B, but not E, produced dose- and endothelium-dependent vasorelaxation. (%maximal relaxation at 5 mg total anthocyanins per liter: Ck = 68 +/- 11, B = 59 +/- 10). Coronary vascular tone, endothelium-dependent vasorelaxation to A23187, and vasorelaxation to DEA NONOate were not affected by exposure of rings to any extract at 0.05 mg total anthocyanins per liter for 5 or 30 min. Ck extract at 0.05 mg total anthocyanins per liter showed the greatest protection against loss of A23187 relaxation following exposure to ROS from pyrogallol (Ck, % maximal relaxation and -logED50 to A23187, respectively, means +/- SE: Ck alone, 93 +/- 5%, 7.91 +/- 0.1; pyrogallol alone, 76 +/- 7%, 7.46 +/- 0.06; pyrogallol + Ck, 98 +/- 1%, 7.82 +/- 0.06; control: 99 +/- 1%, 7.86 +/- 0.07; P < 0.05 control vs. pyrogallol alone). Neither the extracts nor pyrogallol affected responses to DEA NONOate. Thus anthocyanin-enhanced extracts produce endothelium-dependent relaxation in porcine coronary arteries. Extract concentrations too low to directly alter coronary vascular tone protect coronary arteries from ROS without altering vasorelaxation to endogenous or exogenous NO. These results suggest that such extracts could have significant beneficial effects in vascular disease. PMID- 16339349 TI - Basal and evoked levels of bioassayable growth hormone are altered by hindlimb unloading. AB - Bioassayable growth hormone (BGH) in rats is released in large quantities from the pituitary in response to the activation of large, proprioceptive afferent fibers from fast and mixed fiber-type hindlimb musculature. We hypothesized that hindlimb unloading (HU) of adult male rats would 1) reduce the basal levels of plasma BGH, and 2) abolish stimulus-induced BGH release. Rats were exposed to HU for 1, 4, or 8 wk. Plasma and pituitaries were collected under isoflurane anesthesia for hormone analyses. Additionally, at 4 and 8 wk, a subset of rats underwent an in situ electrical stimulation (Stim) of tibial nerve proprioceptive afferents. Basal plasma BGH levels were significantly reduced (-51 and -23%) after 1 and 8 wk of HU compared with ambulatory controls (Amb). Although Amb-Stim rats exhibited increased plasma BGH levels (88 and 143%) and decreased pituitary BGH levels (-27 and -22%) at 4 and 8 wk, respectively, stimulation in HU rats had the opposite effect, reducing plasma BGH (-25 and -33%) and increasing pituitary BGH levels (47 and 10%) relative to HU alone at 4 and 8 wk. The 22-kDa form of GH measured by immunoassay and the plasma corticosterone, T3, T4, and testosterone levels were unchanged by HU or Stim at all time points. These data suggest that BGH synthesis and release from the pituitary are sensitive both to chronically reduced neuromuscular loading and to acute changes in neuromuscular activation, independent of changes in other circulating hormones. Thus BGH may play a role in muscle, bone, and metabolic adaptations that occur in response to chronically unloaded states. PMID- 16339350 TI - Analysis of factors that influence rates of carbon monoxide uptake, distribution, and washout from blood and extravascular tissues using a multicompartment model. AB - To better understand factors that influence carbon monoxide (CO) washout rates, we utilized a multicompartment mathematical model to predict rates of CO uptake, distribution in vascular and extravascular (muscle vs. other soft tissue) compartments, and washout over a range of exposure and washout conditions with varied subject-specific parameters. We fitted this model to experimental data from 15 human subjects, for whom subject-specific parameters were known, multiple washout carboxyhemoglobin (COHb) levels were available, and CO exposure conditions were identical, to investigate the contributions of exposure conditions and individual variability to CO washout from blood. We found that CO washout from venous blood was biphasic and that postexposure times at which COHb samples were obtained significantly influenced the calculated CO half times (P < 0.0001). The first, more rapid, phase of CO washout from the blood reflected the loss of CO to the expired air and to a slow uptake by the muscle compartment, whereas the second, slower washout phase was attributable to CO flow from the muscle compartment back to the blood and removal from blood via the expired air. When the model was used to predict the effects of varying exposure conditions for these subjects, the CO exposure duration, concentration, peak COHb levels, and subject-specific parameters each influenced washout half times. Blood volume divided by ventilation correlated better with half-time predictions than did cardiac output, muscle mass, or ventilation, but it explained only approximately 50% of half-time variability. Thus exposure conditions, COHb sampling times, and individual parameters should be considered when estimating CO washout rates for poisoning victims. PMID- 16339351 TI - Exercise training in normobaric hypoxia in endurance runners. II. Improvement of mitochondrial properties in skeletal muscle. AB - This study investigates whether adaptations of mitochondrial function accompany the improvement of endurance performance capacity observed in well-trained athletes after an intermittent hypoxic training program. Fifteen endurance trained athletes performed two weekly training sessions on treadmill at the velocity associated with the second ventilatory threshold (VT2) with inspired O2 fraction = 14.5% [hypoxic group (Hyp), n = 8] or with inspired O2 fraction = 21% [normoxic group (Nor), n = 7], integrated into their usual training, for 6 wk. Before and after training, oxygen uptake (VO2) and speed at VT2, maximal VO2 (VO2 max), and time to exhaustion at velocity of VO2 max (minimal speed associated with VO2 max) were measured, and muscle biopsies of vastus lateralis were harvested. Muscle oxidative capacities and sensitivity of mitochondrial respiration to ADP (Km) were evaluated on permeabilized muscle fibers. Time to exhaustion, VO2 at VT2, and VO2 max were significantly improved in Hyp (+42, +8, and +5%, respectively) but not in Nor. No increase in muscle oxidative capacity was obtained with either training protocol. However, mitochondrial regulation shifted to a more oxidative profile in Hyp only as shown by the increased Km for ADP (Nor: before 476 +/- 63, after 524 +/- 62 microM, not significant; Hyp: before 441 +/- 59, after 694 +/- 51 microM, P < 0.05). Thus including hypoxia sessions into the usual training of athletes qualitatively ameliorates mitochondrial function by increasing the respiratory control by creatine, providing a tighter integration between ATP demand and supply. PMID- 16339352 TI - Inducibility of male-specific isoforms of cytochrome p450 by sex-dependent growth hormone profiles in hepatocyte cultures from male but not female rats. AB - Although in vivo expression levels of the male-specific hepatic isoforms of cytochrome P450 (P450) (CYP2C11, CYP2C13, CYP2A2, and CYP3A2) are determined by the episodic growth hormone profile secreted by male rats, these isoforms have been completely refractory to growth hormone regulation in hepatocyte culture. By using species-specific rat growth hormone, at subphysiologic in vivo concentrations administered in two daily episodic pulses, we successfully induced CYP2C11 and CYP2A2 to near normal concentrations. Whereas inductive levels of CYP2C13 were subnormal, CYP3A2 was unresponsive to all hormonal treatments, quickly declining to undetectable concentrations. In agreement with in vivo findings, we observed that induction levels of the isoforms were always greatest when the male hepatocytes were exposed to the masculine-like episodic growth hormone profile and least stimulated by the continuous feminine-like hormone profile. When administered alone, dexamethasone consistently increased isoform levels. However, when administered with growth hormone, the glucocorticoid was always antagonistic, suppressing growth hormone induction of CYP2C11, CYP2C13, and CYP2A2. Finally, the P450 isoforms were completely unresponsive to all treatments when the hepatocytes were derived from female rats, supporting earlier findings that expression levels of sexually dimorphic P450 isoforms are inherently irreversible between sexes. PMID- 16339353 TI - Expression of UDP-glucuronosyltransferase isoform mRNAs during inflammation and infection in mouse liver and kidney. AB - Inflammation or infection down-regulates the activity and expression of cytochrome P450 (P450) enzymes involved in hepatic drug clearance, possibly altering drug effectiveness and leading to toxicity. The regulation of UDP glucuronosyltransferases (UGTs) in inflammation and infection is less well characterized. To determine the response of hepatic and renal UGTs during inflammation and infection, mice were administered either saline or 1 mg/kg lipopolysaccharide (LPS) (16 h), or Citrobacter rodentium by oral gavage (6 days). Hepatic mRNA expression of UGT1A1, 1A9, and 2B5 was similarly down regulated after LPS exposure and C. rodentium infection, whereas UGT1A2 and 1A6 mRNAs were unchanged. Effects of C. rodentium infection did not require a functional Toll-like receptor 4. Conversely, renal UGT isoforms were relatively unaffected, except for UGT2B5 induction after LPS treatment. Regulation of UGTs during the inflammatory response exhibits similarities to and differences from regulation of P450s, and may be cytokine-mediated. PMID- 16339355 TI - Digital fluorescence imaging of organic cation transport in freshly isolated rat proximal tubules. AB - The secretion of cationic drugs and endogenous metabolites is a major function of the kidney. This is accomplished by organic cation transport systems, mainly located in the proximal tubules. Here, we describe a model for continuous measurement of organic cation (OC) transport. In this model, organic cation transport in individual freshly isolated rat proximal tubules is investigated by use of digital fluorescence imaging. To directly measure organic cation transport across the basolateral membrane, the fluorescent organic cation 4-(4 dimethylaminostyryl)-N-methylpyridinium (ASP+) is used with a customized perfusion chamber. ASP+ uptake in this model displayed the characteristics of organic cation transport. Over the tested range of 1 to 50 microM, it showed a concentration-dependent uptake across the basolateral membrane. In the presence of competitive inhibitors of OC transport such as N1-methylnicotinamide+, tetraethylammonium+, and choline+, a concentration-dependent and reversible inhibition of ASP+ uptake could be documented. In conclusion, continuous measurement of organic cation transport in freshly isolated rat proximal tubules by digital fluorescence imaging using ASP+ is a useful tool for investigation of drug transport and interactions and, furthermore, may be helpful for investigation of organic cation transport under pathophysiological conditions. PMID- 16339354 TI - Hepatic and renal cytochrome p450 gene regulation during citrobacter rodentium infection in wild-type and toll-like receptor 4 mutant mice. AB - Citrobacter rodentium is the rodent equivalent of human enteropathogenic Escherichia coli infection. This study investigated regulation of hepatic and renal cytochrome P450 (P450) mRNAs, hepatic P450 proteins, cytokines, and acute phase proteins during C. rodentium infection. Female C3H/HeOuJ (HeOu) and C3H/HeJ (HeJ) mice [which lack functional toll-like receptor 4 (TLR4)] were infected with C. rodentium by oral gavage and sacrificed 6 days later. Hepatic CYP4A10 and 4A14 mRNAs were decreased in HeOu mice (<4% of control). CYP3A11, 2C29, 4F14, and 4F15 mRNAs were reduced to 16 to 55% of control levels, whereas CYP2A5, 4F16, and 4F18 mRNAs were induced (180, 190, and 600% of control, respectively). The pattern of P450 regulation in HeJ mice was similar to that in HeOu mice for most P450s, with the exception of the TLR4 dependence of CYP4F15. Hepatic CYP2C, 3A, and 4A proteins in both groups were decreased, whereas CYP2E protein was not. Renal CYP4A10 and 4A14 mRNAs were significantly down-regulated in HeOu mice, whereas other P450s were unaffected. Most renal P450 mRNAs in infected HeJ mice were increased, notably CYP4A10, 4A14, 4F18, 2A5, and 3A13. Hepatic levels of interleukin (IL)-1beta, IL-6, and tumor necrosis factor alpha (TNFalpha) mRNAs were significantly increased in infected HeOu mice, whereas only TNFalpha mRNA was significantly increased in HeJ mice. Hepatic alpha1-acid glycoprotein was induced in both groups, whereas alpha-fibrinogen and angiotensinogen were unchanged. These data indicate that hepatic inflammation induced by C. rodentium infection is mainly TLR4-independent and suggest that hepatic P450 down regulation in this model may be cytokine-mediated. PMID- 16339357 TI - Insights on biology and evolution from microbial genome sequencing. AB - No field of research has embraced and applied genomic technology more than the field of microbiology. Comparative analysis of nearly 300 microbial species has demonstrated that the microbial genome is a dynamic entity shaped by multiple forces. Microbial genomics has provided a foundation for a broad range of applications, from understanding basic biological processes, host-pathogen interactions, and protein-protein interactions, to discovering DNA variations that can be used in genotyping or forensic analyses, the design of novel antimicrobial compounds and vaccines, and the engineering of microbes for industrial applications. Most recently, metagenomics approaches are allowing us to begin to probe complex microbial communities for the first time, and they hold great promise in helping to unravel the relationships between microbial species. PMID- 16339358 TI - Changing perspectives in yeast research nearly a decade after the genome sequence. AB - Research with budding yeast (Saccharomyces cerevisiae) has been transformed by the publication, nearly a decade ago, of the entire genome DNA sequence. The introduction of this first eukaryotic genomic sequence changed the yeast research environment significantly, not just because of dramatic progress in technical means but also because the sequence made accessible a new class of scientific questions. A central goal of yeast research remains the determination of the biological role of every sequence feature in the yeast genome. The most remarkable change has been the shift in perspective from focus on individual genes and functionalities to a more global view of how the cellular networks and systems interact and function together to produce the highly evolved organism we see today. PMID- 16339359 TI - Genomics of the fungal kingdom: insights into eukaryotic biology. AB - The last decade has witnessed a revolution in the genomics of the fungal kingdom. Since the sequencing of the first fungus in 1996, the number of available fungal genome sequences has increased by an order of magnitude. Over 40 complete fungal genomes have been publicly released with an equal number currently being sequenced--representing the widest sampling of genomes from any eukaryotic kingdom. Moreover, many of these sequenced species form clusters of related organisms designed to enable comparative studies. These data provide an unparalleled opportunity to study the biology and evolution of this medically, industrially, and environmentally important kingdom. In addition, fungi also serve as model organisms for all eukaryotes. The available fungal genomic resource, coupled with the experimental tractability of the fungi, is accelerating research into the fundamental aspects of eukaryotic biology. We provide here an overview of available fungal genomes and highlight some of the biological insights that have been derived through their analysis. We also discuss insights into the fundamental cellular biology shared between fungi and other eukaryotic organisms. PMID- 16339360 TI - The Arabidopsis genome: a foundation for plant research. AB - The sequence of the first plant genome was completed and published at the end of 2000. This spawned a series of large-scale projects aimed at discovering the functions of the 25,000+ genes identified in Arabidopsis thaliana (Arabidopsis). This review summarizes progress made in the past five years and speculates about future developments in Arabidopsis research and its implications for crop science. The provision of large populations of gene disruption lines to the research community has greatly accelerated the impact of genomics on many areas of plant science. The tools and community organization required for plant integrative and systems biology approaches are now ready to accomplish the next big step in plant biology--the integration of knowledge and modeling of biological processes. In the future, plant science will continue to be enriched by the alignment of high-quality basic research (generally conducted in Arabidopsis), with strategic objectives in crop plants. The sequence and analysis of an increasing number of crop plant genomes enhance this alignment and provide new insights into genome evolution and crop plant domestication. PMID- 16339361 TI - Grains of knowledge: genomics of model cereals. AB - The economic and scientific importance of the cereals has motivated a rich history of research into their genetics, development, and evolution. The nearly completed sequence of the rice genome is emblematic of a transition to high throughput genomics and computational biology that has also pervaded study of many other cereals. The relatively close (ca. <50 million years old) relationships among morphologically diverse cereals native to environments that sample much of global geographic diversity make the cereals particularly attractive for comparative studies of plant genome evolution. Extensive germplasm resources, largely a byproduct of their economic importance, together with growing collections of defined mutants, provide foundations for a host of post genomic studies to shed more light on the relationship between sequence and function in this important group. Using the rapidly growing capabilities of several informatics resources, genomic data from model cereals are likely to be leveraged tremendously in the study and improvement of a wide range of crop plants that sustain much of the world's population, including many which still lack primary genomic resources. PMID- 16339362 TI - Genomics in C. elegans: so many genes, such a little worm. AB - The Caenorhabditis elegans genome sequence is now complete, fully contiguous telomere to telomere and totaling 100,291,840 bp. The sequence has catalyzed the collection of systematic data sets and analyses, including a curated set of 19,735 protein-coding genes--with >90% directly supported by experimental evidence--and >1300 noncoding RNA genes. High-throughput efforts are under way to complete the gene sets, along with studies to characterize gene expression, function, and regulation on a genome-wide scale. The success of the worm project has had a profound effect on genome sequencing and on genomics more broadly. We now have a solid platform on which to build toward the lofty goal of a true molecular understanding of worm biology with all its implications including those for human health. PMID- 16339363 TI - Drosophila melanogaster: a case study of a model genomic sequence and its consequences. AB - The sequencing and annotation of the Drosophila melanogaster genome, first published in 2000 through collaboration between Celera Genomics and the Drosophila Genome Projects, has provided a number of important contributions to genome research. By demonstrating the utility of methods such as whole-genome shotgun sequencing and genome annotation by a community "jamboree," the Drosophila genome established the precedents for the current paradigm used by most genome projects. Subsequent releases of the initial genome sequence have been improved by the Berkeley Drosophila Genome Project and annotated by FlyBase, the Drosophila community database, providing one of the highest-quality genome sequences and annotations for any organism. We discuss the impact of the growing number of genome sequences now available in the genus on current Drosophila research, and some of the biological questions that these resources will enable to be solved in the future. PMID- 16339364 TI - Unraveling genomic regulatory networks in the simple chordate, Ciona intestinalis. AB - The draft genome of the primitive chordate, Ciona intestinalis, was published three years ago. Since then, significant progress has been made in utilizing Ciona's genomic and morphological simplicity to better understand conserved chordate developmental processes. Extensive annotation and sequencing of staged EST libraries make the Ciona genome one of the best annotated among those that are publicly available. The formation of the Ciona tadpole depends on simple, well-defined cellular lineages, and it is possible to trace the lineages of key chordate tissues such as the notochord and neural tube to the fertilized egg. Electroporation methods permit the targeted expression of regulatory genes and signaling molecules in defined cell lineages, as well as the rapid identification of regulatory DNAs underlying cell-specific gene expression. The recent sequencing of a second Ciona genome (C. savignyi) permits the use of simple alignment algorithms for the identification of conserved noncoding sequences, including microRNA genes and enhancers. Detailed expression profiles are now available for almost every gene that encodes a regulatory protein or cell signaling molecule. The combination of gene-expression profiles, comparative genome analysis, and gene-disruption assays should permit the determination of high-resolution genomic regulatory networks underlying the specification of basic chordate tissues such as the heart, blood, notochord, and neural tube. PMID- 16339365 TI - Fish genomics and biology. AB - The last common ancestor between fish and mammals dates back to the very origin of the vertebrate lineage and today, half of modern vertebrates are fish. It is thus not surprising that several fish species have played important roles in recent years to advance our understanding of vertebrate genome evolution, to inform us on the structure of human genes, and, somewhat more unexpectedly, to provide leads to understanding the function of genes involved in human diseases. Genome sequence comparisons between such distantly related organisms are highly informative due to the accumulation of neutral mutations in nonfunctional regions. Yet humans and fishes share many developmental pathways, organ systems, and physiological mechanisms, making conclusions relevant to human biology. The respective advantages of zebrafish, medaka, Tetraodon, or Takifugu have been well exploited so far with bioinformatics analyses and molecular biology techniques. However the full potential of fish genomics is about to be unleashed with the integration of more traditional disciplines such as biochemistry and physiology, with the study of additional species such as carp, trout, or tilapia and a broadening of its applications to environmental genomics or aquaculture. PMID- 16339366 TI - Xenomics. AB - Xenopus genomics, or Xenomics for short, is coming of age. Indeed, biological insight into processes such as growth factor signaling and patterning of the early embryo is now being gained by combining the value of Xenopus as a model organism for cell and developmental biology with genomic approaches. In this review I address these recent advances and explore future possibilities gained from combining this powerful experimental system with genomic approaches, as well as how our quest to understand basic biological principles will be greatly facilitated though the marriage of Xenopus and genomics. PMID- 16339367 TI - Chicken genome: current status and future opportunities. AB - The chicken genome sequence is important for several reasons. First, the chicken shared a common ancestor with mammals approximately 310 million years ago (Mya) at a phylogenetic distance not previously covered by other genome sequences. It therefore fills a gap in our knowledge and understanding of the evolution and conservation of genes, regulatory sequences, genomes, and karyotypes. The chicken is also a major source of protein in the world, with billions of birds used in meat and egg production each year. It is the first livestock species to be sequenced and so leads the way for others. The sequence and the 2.8 million genetic polymorphisms defined in a parallel project are expected to benefit agriculture and cast new light on animal domestication. Also, as the first bird to be sequenced, it is a model for the 9600 avian species thought to exist today. Many of the features of the chicken genome and its biology make it an ideal organism for studies in development and evolution, along with applications in agriculture and medicine. PMID- 16339368 TI - Advances in livestock genomics: opening the barn door. AB - Genome research in animals used in agriculture has progressed rapidly in recent years, moving from rudimentary genome maps to trait maps to gene discovery. These advances are the result of animal genome projects following closely in the footsteps of the Human Genome Project, which has opened the door to genome research in farm animals. In return, genome research in livestock species is contributing to our understanding of chromosome evolution and to informing the human genome. Enhancement of these contributions plus the much anticipated application of DNA-based tools to animal health and production can be expected as livestock genomics enters its sequencing era. PMID- 16339369 TI - The canine genome. AB - The dog has emerged as a premier species for the study of morphology, behavior, and disease. The recent availability of a high-quality draft sequence lifts the dog system to a new threshold. We provide a primer to use the dog genome by first focusing on its evolutionary history. We overview the relationship of dogs to wild canids and discuss their origin and domestication. Dogs clearly originated from a substantial number of gray wolves and dog breeds define distinct genetic units that can be divided into at least four hierarchical groupings. We review evidence showing that dogs have high levels of linkage disequilibrium. Consequently, given that dog breeds express specific phenotypic traits and vary in behavior and the incidence of genetic disease, genomic-wide scans for linkage disequilibrium may allow the discovery of genes influencing breed-specific characteristics. Finally, we review studies that have utilized the dog to understand the genetic underpinning of several traits, and we summarize genomic resources that can be used to advance such studies. We suggest that given these resources and the unique characteristics of breeds, that the dog is a uniquely valuable resource for studying the genetic basis of complex traits. PMID- 16339370 TI - Impact of genomics on research in the rat. AB - The need to translate genes to function has positioned the rat as an invaluable animal model for genomic research. The significant increase in genomic resources in recent years has had an immediate functional application in the rat. Many of the resources for translational research are already in place and are ready to be combined with the years of physiological knowledge accumulated in numerous rat models, which is the subject of this perspective. Based on the successes to date and the research projects under way to further enhance the infrastructure of the rat, we also project where research in the rat will be in the near future. The impact of the rat genome project has just started, but it is an exciting time with tremendous progress. PMID- 16339371 TI - The mouse genome. AB - The house mouse has been used as a privileged model organism since the early days of genetics, and the numerous experiments made with this small mammal have regularly contributed to enrich our knowledge of mammalian biology and pathology, ranging from embryonic development to metabolic disease, histocompatibility, immunology, behavior, and cancer. Over the past two decades, a number of large scale integrated and concerted projects have been undertaken that will probably open a new era in the genetics of the species. The sequencing of the genome, which will allow researchers to make comparisons with other mammals and identify regions conserved by evolution, is probably the most important project, but many other initiatives, such as the massive production of point or chromosomal mutations associated with comprehensive and standardized phenotyping of the mutant phenotypes, will help annotation of the approximately 25,000 genes packed in the mouse genome. In the same way, and as another consequence of the sequencing, the discovery of many single nucleotide polymorphisms and the development of new tools and resources, like the Collaborative Cross, will contribute to the development of modern quantitative genetics. It is clear that mouse genetics has changed dramatically over the last 10-15 years and its future looks promising. PMID- 16339372 TI - Genomics of the future: identification of quantitative trait loci in the mouse. AB - Positional cloning of quantitative trait loci in rodents is a common approach to identify genes involved in complex phenotypes, including genes important to human disease. However, cloning the causative genes has proved to be more difficult than determining their positions. New tools such as genomic sequence, clone libraries, and new genomic-based methods offer new approaches to identify these genes. Here we review how these new tools and approaches will improve our ability to discover the genes important in complex traits. PMID- 16339373 TI - Comparing the human and chimpanzee genomes: searching for needles in a haystack. AB - The chimpanzee genome sequence is a long-awaited milestone, providing opportunities to explore primate evolution and genetic contributions to human physiology and disease. Humans and chimpanzees shared a common ancestor approximately 5-7 million years ago (Mya). The difference between the two genomes is actually not approximately 1%, but approximately 4%--comprising approximately 35 million single nucleotide differences and approximately 90 Mb of insertions and deletions. The challenge is to identify the many evolutionarily, physiologically, and biomedically important differences scattered throughout these genomes while integrating these data with emerging knowledge about the corresponding "phenomes" and the relevant environmental influences. It is logical to tackle the genetic aspects via both genome-wide analyses and candidate gene studies. Genome-wide surveys could eliminate the majority of genomic sequence differences from consideration, while simultaneously identifying potential targets of opportunity. Meanwhile, candidate gene approaches can be based on such genomic surveys, on genes that may contribute to known differences in phenotypes or disease incidence/severity, or on mutations in the human population that impact unique aspects of the human condition. These two approaches will intersect at many levels and should be considered complementary. We also cite some known genetic differences between humans and great apes, realizing that these likely represent only the tip of the iceberg. PMID- 16339374 TI - Structure and function of the human genome. AB - The human genome project has had an impact on both biological research and its political organization; this review focuses primarily on the scientific novelty that has emerged from the project but also touches on its political dimensions. The project has generated both anticipated and novel information; in the later category are the description of the unusual distribution of genes, the prevalence of non-protein-coding genes, and the extraordinary evolutionary conservation of some regions of the genome. The applications of the sequence data are just starting to be felt in basic, rather than therapeutic, biomedical research and in the vibrant human origins and variation debates. The political impact of the project is in the unprecedented extent to which directed funding programs have emerged as drivers of basic research and the organization of the multidisciplinary groups that are needed to utilize the human DNA sequence. PMID- 16339375 TI - Emerging technologies in DNA sequencing. AB - Demand for DNA sequence information has never been greater, yet current Sanger technology is too costly, time consuming, and labor intensive to meet this ongoing demand. Applications span numerous research interests, including sequence variation studies, comparative genomics and evolution, forensics, and diagnostic and applied therapeutics. Several emerging technologies show promise of delivering next-generation solutions for fast and affordable genome sequencing. In this review article, the DNA polymerase-dependent strategies of Sanger sequencing, single nucleotide addition, and cyclic reversible termination are discussed to highlight recent advances and potential challenges these technologies face in their development for ultrafast DNA sequencing. PMID- 16339376 TI - Genome annotation past, present, and future: how to define an ORF at each locus. AB - Driven by competition, automation, and technology, the genomics community has far exceeded its ambition to sequence the human genome by 2005. By analyzing mammalian genomes, we have shed light on the history of our DNA sequence, determined that alternatively spliced RNAs and retroposed pseudogenes are incredibly abundant, and glimpsed the apparently huge number of non-coding RNAs that play significant roles in gene regulation. Ultimately, genome science is likely to provide comprehensive catalogs of these elements. However, the methods we have been using for most of the last 10 years will not yield even one complete open reading frame (ORF) for every gene--the first plateau on the long climb toward a comprehensive catalog. These strategies--sequencing randomly selected cDNA clones, aligning protein sequences identified in other organisms, sequencing more genomes, and manual curation--will have to be supplemented by large-scale amplification and sequencing of specific predicted mRNAs. The steady improvements in gene prediction that have occurred over the last 10 years have increased the efficacy of this approach and decreased its cost. In this Perspective, I review the state of gene prediction roughly 10 years ago, summarize the progress that has been made since, argue that the primary ORF identification methods we have relied on so far are inadequate, and recommend a path toward completing the Catalog of Protein Coding Genes, Version 1.0. PMID- 16339377 TI - Hotspots of mutation and breakage in dog and human chromosomes. AB - Sequencing of the dog genome allows an investigation of the location-dependent evolutionary processes that occurred since the common ancestor of primates and carnivores, approximately 95 million years ago. We investigated variations in G+C nucleotide fraction and synonymous nucleotide substitution rates (Ks) across dog and human genomes. Our results show that dog genes located either in subtelomeric and pericentromeric regions, or in short synteny blocks, possess significantly elevated G+C fraction and Ks values. Human subtelomeric, but not pericentromeric, genes also exhibit these elevations. We then examined 1.048 Gb of human sequence that is likely not to have been located near a primate telomere at any time since the common ancestor of dog and human. We observed that regions of highest G+C or Ks ("hotspots"; median sizes of 0.5 or 1.3 Mb, respectively) within this sequence were preferentially segregated to dog subtelomeres and pericentromeres during the rearrangements that eventually gave rise to the extant canine karyotype. Our data cannot be accounted for solely on the basis of gradually elevating G+C fractions in subtelomeric regions as a consequence of biased gene conversion. Rather, we propose that high G+C sequences are found preferentially within dog subtelomeres as a direct consequence of chromosomal fission occurring more frequently within regions elevated in G+C. PMID- 16339378 TI - Short interspersed elements (SINEs) are a major source of canine genomic diversity. AB - SINEs are retrotransposons that have enjoyed remarkable reproductive success during the course of mammalian evolution, and have played a major role in shaping mammalian genomes. Previously, an analysis of survey-sequence data from an individual dog (a poodle) indicated that canine genomes harbor a high frequency of alleles that differ only by the absence or presence of a SINEC_Cf repeat. Comparison of this survey-sequence data with a draft genome sequence of a distinct dog (a boxer) has confirmed this prediction, and revealed the chromosomal coordinates for >10,000 loci that are bimorphic for SINEC_Cf insertions. Analysis of SINE insertion sites from the genomes of nine additional dogs indicates that 3%-5% are absent from either the poodle or boxer genome sequences--suggesting that an additional 10,000 bimorphic loci could be readily identified in the general dog population. We describe a methodology that can be used to identify these loci, and could be adapted to exploit these bimorphic loci for genotyping purposes. Approximately half of all annotated canine genes contain SINEC_Cf repeats, and these elements are occasionally transcribed. When transcribed in the antisense orientation, they provide splice acceptor sites that can result in incorporation of novel exons. The high frequency of bimorphic SINE insertions in the dog population is predicted to provide numerous examples of allele-specific transcription patterns that will be valuable for the study of differential gene expression among multiple dog breeds. PMID- 16339379 TI - Selective sweep mapping of genes with large phenotypic effects. AB - Many domestic dog breeds have originated through fixation of discrete mutations by intense artificial selection. As a result of this process, markers in the proximity of genes influencing breed-defining traits will have reduced variation (a selective sweep) and will show divergence in allele frequency. Consequently, low-resolution genomic scans can potentially be used to identify regions containing genes that have a major influence on breed-defining traits. We model the process of breed formation and show that the probability of two or three adjacent marker loci showing a spurious signal of selection within at least one breed (i.e., Type I error or false-positive rate) is low if highly variable and moderately spaced markers are utilized. We also use simulations with selection to demonstrate that even a moderately spaced set of highly polymorphic markers (e.g., one every 0.8 cM) has high power to detect regions targeted by strong artificial selection in dogs. Further, we show that a gene responsible for black coat color in the Large Munsterlander has a 40-Mb region surrounding the gene that is very low in heterozygosity for microsatellite markers. Similarly, we survey 302 microsatellite markers in the Dachshund and find three linked monomorphic microsatellite markers all within a 10-Mb region on chromosome 3. This region contains the FGFR3 gene, which is responsible for achondroplasia in humans, but not in dogs. Consequently, our results suggest that the causative mutation is a gene or regulatory region closely linked to FGFR3. PMID- 16339380 TI - Interaction between the X chromosome and an autosome regulates size sexual dimorphism in Portuguese Water Dogs. AB - Size sexual dimorphism occurs in almost all mammals. In Portuguese Water Dogs, much of the difference in skeletal size between females and males is due to the interaction between a Quantitative Trait Locus (QTL) on the X-chromosome and a QTL linked to Insulin-like Growth Factor 1 (IGF-1) on the CFA 15 autosome. In females, the haplotype of CFA 15 resulting in small size is dominant. In males, the haplotype for large size is dominant. Females, homozygous at the CHM marker on the X chromosome and homozygous for the large size CFA 15 haplotype are, on average, as large as large males. However, all females that are heterozygous at the CHM marker are small, regardless of their CFA 15 genotype. This interaction suggests a genetic mechanism that in turn leads to a scenario for the evolution of size sexual dimorphism consistent with a proposal of Lande that sexual dimorphism can evolve because females secondarily become smaller than males as a consequence of natural selection for optimal size. Our results also can explain Rensch's Rule, which states that size is often positively correlated with the level of size sexual dimorphism. PMID- 16339381 TI - Genetics of canid skeletal variation: size and shape of the pelvis. AB - The mammalian skeleton presents an ideal system in which to study the genetic architecture of a set of related polygenic traits and the skeleton of the domestic dog (Canis familiaris) is arguably the best system in which to address the relationship between genes and anatomy. We have analyzed the genetic basis for skeletal variation in a population of >450 Portuguese Water Dogs. At this stage of this ongoing project, we have identified >40 putative quantitative trait loci (QTLs) for heritable skeletal phenotypes located on 22 different chromosomes, including the "X." A striking aspect of these is the regulation of suites of traits representing bones located in different parts of the skeleton but related by function. Here we illustrate this by describing genetic variation in postcranial morphology. Two suites of traits are involved. One regulates the size of the pelvis relative to dimensions of the limb bones. The other regulates the shape of the pelvis. Both are examples of trade-offs that may be prototypical of different breeds. For the size of the pelvis relative to limb bones, we describe four QTLs located on autosome CFA 12, 30, 31, and X. For pelvic shape we describe QTLs on autosome CFA 2, 3, 22, and 36. The relation of these polygenic systems to musculoskeletal function is discussed. PMID- 16339382 TI - Construction of a 2-Mb resolution BAC microarray for CGH analysis of canine tumors. AB - Recognition of the domestic dog as a model for the comparative study of human genetic traits has led to major advances in canine genomics. The pathophysiological similarities shared between many human and dog diseases extend to a range of cancers. Human tumors frequently display recurrent chromosome aberrations, many of which are hallmarks of particular tumor subtypes. Using a range of molecular cytogenetic techniques we have generated evidence indicating that this is also true of canine tumors. Detailed knowledge of these genomic abnormalities has the potential to aid diagnosis, prognosis, and the selection of appropriate therapy in both species. We recently improved the efficiency and resolution of canine cancer cytogenetics studies by developing a small-scale genomic microarray comprising a panel of canine BAC clones representing subgenomic regions of particular interest. We have now extended these studies to generate a comprehensive canine comparative genomic hybridization (CGH) array that comprises 1158 canine BAC clones ordered throughout the genome with an average interval of 2 Mb. Most of the clones (84.3%) have been assigned to a precise cytogenetic location by fluorescence in situ hybridization (FISH), and 98.5% are also directly anchored within the current canine genome assembly, permitting direct translation from cytogenetic aberration to DNA sequence. We are now using this resource routinely for high-throughput array CGH and single-locus probe analysis of a range of canine cancers. Here we provide examples of the varied applications of this resource to tumor cytogenetics, in combination with other molecular cytogenetic techniques. PMID- 16339383 TI - Vasopressin induces depolarization and state-dependent firing patterns in rat thalamic paraventricular nucleus neurons in vitro. AB - The thalamic midline paraventricular nucleus (PVT) is prominently innervated by vasopressin-immunoreactive neurons from the suprachiasmatic nucleus (SCN), site of the brain's biological clock. Using patch-clamp recordings in slice preparations taken from Wistar rats during the subjective day, we examined 90 PVT neurons for responses to bath-applied AVP (0.5-2 microM; 1-3 min). In current clamp at resting membrane potentials (-65 +/- 1 mV), PVT neurons displayed low threshold spikes (LTSs) and burst firing patterns. In 50% of cells tested, AVP induced a slowly rising, prolonged membrane depolarization and tonic firing, returning to burst firing upon recovery. AVP modulated hyperpolarization activated LTSs by decreasing the time to the initial sodium spike at the onset of LTS, also increasing the duration of the afterdepolarization. Responses were blockable with a V(1a) receptor antagonist (Manning compound). Under voltage clamp, AVP induced a TTX-resistant, slowly rising, and prolonged (approximately 15 min) inward current (<40 pA). Current-voltage relationship (I-V) analyses of the AVP responses revealed a decrease in membrane conductance to 73.1 +/- 6.2% of control, with net AVP current reversing at -106 +/- 4 mV, and decreased inward rectification at negative potentials. These observations are consistent with an AVP-induced closure of an inwardly rectifying potassium conductance. On the basis of these in vitro observations, we suggest that the SCN vasopressinergic innervation of PVT is excitatory in nature, possibly releasing AVP with circadian rhythmicity and contributing to state-dependent firing patterns in PVT neurons over the sleep-wake cycle. PMID- 16339384 TI - Altered circadian rhythm reentrainment to light phase shifts in rats with low levels of brain angiotensinogen. AB - In this study, we aimed to investigate the adaptation of blood pressure (BP), heart rate (HR), and locomotor activity (LA) circadian rhythms to light cycle shift in transgenic rats with a deficit in brain angiotensin [TGR(ASrAOGEN)]. BP, HR, and LA were measured by telemetry. After baseline recordings (bLD), the light cycle was inverted by prolonging the light by 12 h and thereafter the dark period by 12 h, resulting in inverted dark-light (DL) or light-dark (LD) cycles. Toward that end, a 24-h dark was maintained for 14 days (free-running conditions). When light cycle was changed from bLD to DL, the acrophases (peak time of curve fitting) of BP, HR, and LA shifted to the new dark period in both SD and TGR(ASrAOGEN) rats. However, the readjustment of the BP and HR acrophases in TGR(ASrAOGEN) rats occurred significantly slower than SD rats. The LA acrophases changed similarly in both strains. When light cycle was changed from DL to LD by prolonging the dark period by 12 h, the reentrainment of BP and LA occurred faster than the previous shift in both strains. The readjustment of the BP and HR acrophases in TGR(ASrAOGEN) rats occurred significantly slower than SD rats. In free-running conditions, the circadian rhythms of the investigated parameters adapted in TGR(ASrAOGEN) and SD rats in a similar manner. These results demonstrate that the brain RAS plays an important role in mediating the effects of light cycle shifts on the circadian variation of BP and HR. The adaptive behavior of cardiovascular circadian rhythms depends on the initial direction of light-dark changes. PMID- 16339385 TI - Novel quantitative phenotypes of exercise training in mouse models. AB - Regular physical exercise has beneficial effects in many human disease states, including cardiovascular diseases, cancer, and depression. Exercise training of genetically modified mouse models may provide insight into the molecular mechanisms that underlie the beneficial effects of exercise. Presently, there is relatively little understanding of the normal physiology of mouse exercise. In this paper, we describe a novel computerized voluntary wheel-running system capable of recording and analyzing individual wheel rotations. Using this system, we demonstrate that C57BL/6 mice run considerable distances during the night in short bouts and at a preferred speed: the cruising speed. We find that the vast majority of running occurs around this cruising speed, which is close to the maximum speed at which the animal can run but is significantly higher than the average speeds recorded by simple digital odometers. We describe how these parameters vary with exercise training and demonstrate marked sex differences in the patterns of voluntary exercise. The results of this study have important implications for the design and interpretation of both voluntary and forced exercise experiments in mouse models. The novel parameters described provide more physiological quantitative measures of voluntary exercise activity and training and will extend the physiological utility of exercise training as a phenotyping tool in genetic mouse models. PMID- 16339386 TI - Modulation of single-nephron GFR in the db/db mouse model of type 2 diabetes mellitus. AB - Hyperfiltration has been implicated in the progression toward diabetic nephropathy in type 2 diabetes mellitus (DM2). This study focuses for the first time on the in vivo modulation of single-nephron GFR (SNGFR) in the classic B6.Cg m(+/+)Lepr(db)/J (db/db) mouse model of DM2. To obtain stable preparations, it was necessary to use a sustaining infusion of 3.3 ml.100 g body wt(-1) x h(-1), or higher. SNGFR (measured both proximally and distally) was greater in db/db vs. heterozygote (db/m) mice (P < 0.05) but not vs. the wild-type (WT) mice. The tubuloglomerular feedback (TGF) responses, determined as free-flow proximal vs. distal SNGFR differences, were significant in db/db mice (11.6 +/- 0.8 vs. 9.3 +/ 1.0 nl/min, P < 0.01), in db/m mice (8.0 +/- 0.8 vs. 7.2 +/- 0.6 nl/min, P < 0.02), and WT mice (9.9 +/- 0.6 vs. 8.9 +/- 0.7 nl/min, P < 0.05). After increasing the sustaining infusion in the db/db mice, to offset glycosuric urine losses, the SNGFR increased significantly, and the TGF response was abolished. In these volume-replete db/db mice, absolute fluid reabsorption measured both at the late proximal and distal tubular sites were significantly increased vs. db/m mice infused at 3.3 ml.100 g body wt(-1) x h(-1). After infusion of the neuronal nitric oxide synthase (nNOS) inhibitor S-methylthiocitrulline, SNGFR fell in both db/db and db/m mice. These studies show that SNGFR is elevated in this mouse model of DM2, is suppressed by nNOS inhibition, and is modulated by TGF influences that are altered by the diabetic state and responsive to changes in extracellular fluid volume. PMID- 16339387 TI - Sepsis and inflammatory insults downregulate IGFBP-5, but not IGFBP-4, in skeletal muscle via a TNF-dependent mechanism. AB - The purpose of the present study was to determine whether catabolic stimuli that induce muscle atrophy alter the muscle mRNA abundance of insulin-like growth factor binding protein (IGFBP)-4 and -5, and if so determine the physiological mechanism for such a change. Catabolic insults produced by endotoxin (LPS) and sepsis decreased IGFBP-5 mRNA time- and dose-dependently in gastrocnemius muscle. This reduction did not result from muscle disuse because hindlimb immobilization increased IGFBP-5. Continuous infusion of a nonlethal dose of tumor necrosis factor-alpha (TNF-alpha) decreased IGFBP-5 mRNA 70%, whereas pretreatment of septic rats with a neutralizing TNF binding protein completely prevented the reduction in muscle IGFBP-5. The addition of LPS or TNF-alpha to cultured C(2)C(12) myoblasts also decreased IGFBP-5 expression. Although exogenously administered growth hormone (GH) increased IGFBP-5 mRNA 2-fold in muscle from control rats, muscle from septic animals was GH resistant and no such elevation was detected. In contrast, exogenous administration of IGF-I as part of a binary complex composed of IGF-I/IGFBP-3 produced comparable increases in IGFBP-5 mRNA in both control and septic muscle. Concomitant determinations of IGF-I mRNA content revealed a positive linear relationship between IGF-I and IGFBP-5 mRNA in the same muscle in response to LPS, sepsis, TNF-alpha, and GH treatment. Although dexamethasone decreased muscle IGFBP-5, pretreatment of rats with the glucocorticoid receptor antagonist RU486 did not prevent the sepsis-induced decrease in IGFBP-5 mRNA. In contrast, muscle IGFBP-4 mRNA abundance was not significantly altered by LPS, sepsis, or hindlimb immobilization. In summary, these data demonstrate that various inflammatory insults decrease muscle IGFBP-5 mRNA, without altering IGFBP-4, by a TNF-dependent glucocorticoid-independent mechanism. Finally, IGF-I appears to be a dominant positive regulator of IGFBP-5 gene expression in muscle under both normal and catabolic conditions. PMID- 16339388 TI - Different metabolic responses to central and peripheral injection of enterostatin. AB - Enterostatin, a pentapeptide cleaved from procolipase, suppresses fat intake after peripheral and central administration. Chronic treatment of rats with enterostatin decreases body weight and body fat. The effect was greater than could be accounted by the reduction in food intake alone. Hence, we have investigated the effect of enterostatin on energy metabolism. Male Sprague-Dawley rats adapted to a high-fat diet were implanted with lateral cerebral ventricular or amygdala cannulas. The metabolic effects were determined by indirect calorimetry. After habituation to the test cages, fasted rats were injected with either saline vehicle or enterostatin given either intraperitoneally (100 nmol) or intracerebroventricularly (1 nmol) or into specific brain regions [amygdala (0.01 nmol) or paraventricular nucleus (PVN) (0.1 nmol)]. Respiratory quotient (RQ) and energy expenditure were monitored over 2 h. Intraperitoneal enterostatin reduced RQ (saline: 0.81 +/- 0.02 vs. enterostatin: 0.76 +/- 0.01) and increased energy expenditure by 44%. Intracerebroventricular enterostatin increased the energy expenditure without any effects on RQ, whereas PVN enterostatin increased metabolic rate, while preventing the increase in RQ observed in the control animals. In contrast, neither RQ nor energy expenditure was altered after enterostatin was injected into the amygdala. Enterostatin activated AMP-activated protein kinase in primary cultures of human myocytes in a dose- and time dependent manner and increased the rate of fatty acid beta-oxidation. These findings suggest that enterostatin regulates energy expenditure and substrate partitioning through both peripheral and central effects. PMID- 16339389 TI - Inactivation of the pure antiestrogen fulvestrant and other synthetic estrogen molecules by UDP-glucuronosyltransferase 1A enzymes expressed in breast tissue. AB - Fulvestrant (Faslodex) is administered by intramuscular injection and is converted into ketone, sulfate, sulfone and glucuronide metabolites. Glucuronidation, catalyzed by 18 members of the UDP-glucuronosyltransferase (UGT) enzyme family, plays a major role in the elimination of natural estrogens. The present study was aimed at identifying and characterizing human UGT enzymes involved in the glucuronidation of this antiestrogen as well as other synthetic estrogen derivatives with aliphatic chains on the E2 molecule. In contrast to E2, which is conjugated by UGT1A1, -1A3, -1A8, -1A10, and -2B7, fulvestrant is glucuronidated by UGT1A1, -1A3, -1A4, and -1A8. The four UGT1A-fulvestrant conjugating enzymes glucuronidate this substrate at position 3, whereas only UGT1A8 also produces fulvestrant-17-glucuronide. For E2, only UGT1A3 and UGT2B7 are capable to conjugate at 17-hydroxyposition. These observations indicate that addition of an aliphatic chain to the E2 molecule modifies the specificity of the UGT enzymes toward the C18 molecules. To further investigate the specificity of these enzymes, a series of E2 derivatives with aliphatic or phenyl chains at position 2, 7alpha, and 11beta was also tested for its conjugation with human UGT enzymes. It was observed that, in addition to UGT1A3, UGT1A1 and UGT1A8 also played important roles for the glucuronidation of these compounds. This suggests that the basic structure of E2 is one of the major determinants for the glucuronidation catalyzed by this group of enzymes. Considering the high level of UGT1A3 and -1A4 expression in the gastrointestinal tract and mammary gland, our results suggest that fulvestrant can be inactivated both in intestine and in its target tissue. PMID- 16339390 TI - Vascular adhesion protein-1 plays an important role in postischemic inflammation and neuropathology in diabetic, estrogen-treated ovariectomized female rats subjected to transient forebrain ischemia. AB - Endothelial vascular adhesion protein-1 (VAP-1) facilitates leukocyte adhesion and infiltration. This relates partly to the function of VAP-1 as a semicarbazide sensitive amine oxidase (SSAO). We examined the effects of VAP-1/SSAO inhibition [via LJP-1207 (N'-(2-phenyl-allyl)-hydrazine hydrochloride)] on pial venular leukocyte adhesion and infiltration (at 2-10 h of reperfusion) and neuropathology (at 72 h of reperfusion) after transient forebrain ischemia (TFI). A model associated with increased postischemic inflammation was used-i.e., diabetic ovariectomized (OVX) female rats given chronic estrogen replacement therapy (ERT). We compared rats treated, either at the onset or at 6 h of reperfusion, with saline or LJP-1207. Additional rats, rendered neutropenic 24 h before TFI, were studied. In saline-treated controls, intravascular accumulation of adherent leukocytes gradually increased, reaching 15 to 20% of the venular area, at which point neutrophil infiltration commenced (at approximately 6 h). In the rats given LJP-1207 at the onset of reperfusion, limited neutrophil adhesion ( approximately 5% maximum) and no infiltration were observed. These results generally paralleled those in neutropenic rats. In rats treated at 6 h of reperfusion, the pattern of neutrophil adhesion was similar to that of the saline-treated group up to 6 h, but further infiltration was essentially prevented. Neurologic outcomes and histopathology were similar to one another in the LJP-1207-treated and neutropenic groups and significantly improved over those in saline-treated controls. Thus, VAP-1-mediated post-TFI leukocyte adhesion/infiltration in diabetic OVX females given chronic ERT contributes substantially to neuropathology. One implication is that specifically preventing leukocyte infiltration provides a substantial measure of neuroprotection. This could explain the finding of LJP-1207 having at least a 6-h therapeutic window in this model. PMID- 16339391 TI - Tibolone metabolism in human liver is catalyzed by 3alpha/3beta-hydroxysteroid dehydrogenase activities of the four isoforms of the aldo-keto reductase (AKR)1C subfamily. AB - Tibolone [[7alpha,17alpha]-17-hydroxy-7-methyl-19-norpregn-5(10)-en-20-yn-3-one] is used to treat climacteric symptoms and prevent osteoporosis. It exerts tissue selective effects via site-specific metabolism into 3alpha- and 3beta hydroxymetabolites and a Delta4-isomer. Recombinant human cytosolic aldo-keto reductases 1C1 and 1C2 (AKR1C1 and AKR1C2) produce 3beta-hydroxytibolone, and the liver-specific AKR1C4 produces predominantly 3alpha-hydroxytibolone. These observations may account for the appearance of 3beta-hydroxytibolone in target tissues and 3alpha-hydroxytibolone in the circulation. Using liver autopsy samples (which express AKR1C1-AKR1C4), tibolone was reduced via 3alpha- and 3beta hydroxysteroid dehydrogenase (HSD) activity. 3beta-Hydroxytibolone was exclusively formed in the cytosol and was inhibited by the AKR1C2-specific inhibitor 5beta-cholanic acid-3alpha, 7alpha-diol. The cytosolic formation of 3alpha-hydroxytibolone was inhibited by an AKR1C4-selective inhibitor, phenolphthalein. The ratio of these stereoisomers was 4:1 in favor of 3beta hydroxytibolone. In HepG2 cell cytosol and intact cells (which do not express AKR1C4), tibolone was exclusively reduced to 3beta-hydroxytibolone and was blocked by the AKR1C1-AKR1C3 inhibitor flufenamic acid. In primary hepatocytes (which express AKR1C1-AKR1C4), time-dependent reduction of tibolone into 3beta- and 3alpha-hydroxytibolone was observed again in a 4:1 ratio. 3beta-HSD activity was inhibited by both 5beta-cholanic acid-3alpha,7alpha-diol and flufenamic acid, implicating a role for AKR1C2 and AKR1C1. By contrast, the formation of 3alpha hydroxytibolone was exclusively inhibited by phenolphthalein implicating AKR1C4 in this reaction. 3beta- and 3alpha-Hydroxytibolone were rapidly metabolized into polar metabolites (>85%). The formation of minor amounts of tibolone was also observed followed by AKR1C-catalyzed epimerization. The low hepatic formation of 3alpha-hydroxytibolone suggests that AKR1C4 is not the primary source of this metabolite and instead it maybe formed by an intestinal or enterobacterial 3alpha HSD. PMID- 16339392 TI - Induction of renal 20-hydroxyeicosatetraenoic acid by clofibrate attenuates high fat diet-induced hypertension in rats. AB - This study compared renal hemodynamics, the expression of CYP4A isoforms [the enzymes for 20-hydroxyeicosatetraenoic acid (20-HETE) production], and tubular sodium transporters in male rats fed a high-fat (HF) or control diet for 10 weeks. We also studied the effect of treatment with clofibrate, a CYP4A inducer, on sodium retention and renal function and on CYP4A expression in HF rats. HF rats had higher blood pressure (BP), renal plasma flow, and glomerular filtration rate (GFR), but no significant change in renal vascular resistance. Reverse transcription-polymerase chain reaction analysis showed that CYP4A1 and CYP4A8 expression was significantly decreased in the renal cortex of HF rats. Western blot analysis showed up-regulation of expression of the alpha-subunit of the epithelial sodium channel (alpha-ENaC), the beta-subunit of the epithelial sodium channel (beta-ENaC), sodium/hydrogen exchanger (NHE)-3, and the renal outer medulla K(+) channel (ROMK) in HF rats, whereas expression of the gamma-subunit of the epithelial sodium channel and the alpha1-subunit of Na(+)-K(+)-ATPase remained unchanged. Thus, HF treatment caused the reduction of renal CYP4A1 and CYP4A8 expression, whereas the increases in alpha-ENaC, beta-ENaC, NHE-3, and ROMK expression in renal tubules may have contributed sodium retention and hypertension in HF rats. Furthermore, clofibrate treatment (240 mg/kg/day) caused the decrease of BP and GFR and the attenuation of cumulative sodium balance in HF rats. The attenuation of sodium retention by clofibrate treatment is linked to decreased expression of NHE-3 in renal cortex. Clofibrate induction of CYP4A expression occurred in proximal tubules and in the thick ascending limb of the loop of Henle but not in renal microvessels. This induction correlated with the expression of peroxisome proliferator-activated receptor (PPARalpha) in renal tubules. Therefore, these results suggest that the effects of clofibrate on sodium retention and blood pressure regulation in HF rats may be due to the induction of renal tubular 20-HETE production through the PPARalpha pathway. PMID- 16339393 TI - Modeling drug- and system-related changes in body temperature: application to clomethiazole-induced hypothermia, long-lasting tolerance development, and circadian rhythm in rats. AB - The aim of the present investigation was to develop a pharmacokinetic pharmacodynamic model for the characterization of clomethiazole (CMZ)-induced hypothermia and the rapid development of long-lasting tolerance in rats while taking into account circadian rhythm in baseline and the influence of handling. CMZ-induced hypothermia and tolerance was measured using body temperature telemetry in male Sprague-Dawley rats, which were given s.c. bolus injections of 0, 15, 150, 300, and 600 micromol kg(-1) and 24-h s.c. continuous infusions of 0, 20, and 40 micromol kg(-1) h(-1) using osmotic pumps. The duration of tolerance was studied by repeated injections of 300 micromol kg(-1) at 3- to 32-day intervals. Plasma exposure to CMZ was obtained in satellite groups of catheterized rats. Fitted population concentration-time profiles served as input for the pharmacodynamic analysis. The asymmetric circadian rhythm in baseline body temperature was successfully described by a novel negative feedback model incorporating external light-dark conditions. An empirical function characterized the transient increase in temperature upon handling of the animal. A feedback model for temperature regulation and tolerance development allowed estimation of CMZ potency at 30 +/- 1 microM. The delay in onset of tolerance was estimated via a series of four transit compartments at 7.6 +/- 2 h. The long-lasting tolerance was assumed to be caused by inactivation of a mediator with an estimated turnover time of 46 +/- 3 days. This multicomponent turnover model was able to quantify the CMZ-induced hypothermia, circadian rhythm in baseline, and rapid onset of a long-lasting tolerance to CMZ in rats. PMID- 16339394 TI - Peripherally mediated antinociception of the mu-opioid receptor agonist 2 [(4,5alpha-epoxy-3-hydroxy-14beta-methoxy-17-methylmorphinan-6beta yl)amino]acetic acid (HS-731) after subcutaneous and oral administration in rats with carrageenan-induced hindpaw inflammation. AB - Opioids induce analgesia by activating opioid receptors not only within the central nervous system but also on peripheral sensory neurons. This study investigated peripherally mediated antinociception produced by the mu-opioid receptor agonist 2-[(4,5alpha-epoxy-3-hydroxy-14beta-methoxy-17-methylmorphinan 6beta-yl)amino]acetic acid (HS-731) after s.c. and oral administration in rats with carrageenan-induced hindpaw inflammation. Antinociceptive effects after s.c. administration were assessed 3 h after intraplantar carrageenan injection and compared with those of centrally acting mu-opioid agonists 14-methoxymetopon and morphine. Opioid agonists caused dose-dependent increases in inflamed paw withdrawal latencies to mechanical and thermal stimulation. The time course of action was different, in that HS-731 (20 microg/kg s.c.) produced significant long-lasting effects up to 4 h after administration, whereas 14-methoxymetopon (20 microg/kg) and morphine (2 mg/kg) reached their peak of action at 10 to 30 min, and their effect declined rapidly thereafter. Subcutaneous administration of the peripherally selective opioid antagonist naloxone methiodide inhibited antinociception elicited by HS-731 (20 microg/kg s.c.), whereas it was ineffective against 14-methoxymetopon (20 microg/kg s.c.). Moreover, the antinociception produced by 100 microg/kg s.c. HS-731 was dose-dependently reversed by s.c. naloxone methiodide. This indicates that HS-731 preferentially activates peripheral opioid receptors, whereas 14-methoxymetopon mediates analgesia via central mechanisms. Orally administered HS-731 significantly reduced hyperalgesia in the inflamed paw induced by carrageenan, which was reversible by s.c. administered naloxone methiodide. These results show that systemic (s.c. and oral) treatment with the mu-opioid agonist HS-731 produces potent and long-lasting antinociception through peripheral mechanisms in rats with carrageenan-induced hindpaw inflammation. PMID- 16339395 TI - Comparative cue generalization profiles of L-838, 417, SL651498, zolpidem, CL218,872, ocinaplon, bretazenil, zopiclone, and various benzodiazepines in chlordiazepoxide and zolpidem drug discrimination. AB - The zolpidem discriminative cue is mediated by GABA(A)-alpha1 receptors, whereas the chlordiazepoxide cue may be mediated via non-alpha1 GABA(A) receptors because compounds with selective affinity for GABA(A)-alpha1 receptors fully generalize to the former cue. We predicted that L-838,417 [7-tert-butyl-3-(2,5 difluorophenyl)-6-(2-methyl-2H-1,2,4-triazol-3-ylmethoxy)-1,2,4-triazolo[4,3 b]pyridazine], a partial agonist at non-alpha1 GABA(A) receptors and an antagonist at GABA(A)-alpha1 receptors, would generalize to the chlordiazepoxide but not the zolpidem-discriminative cue. SL651498 [6-fluoro-9-methyl-2-phenyl-4 (pyrrolidin-1-yl-carbonyl)-2,9-dihydro-1H-pyridol[3,4-b]indol-1-one] is a full agonist at GABA(A)-alpha2 receptors, with lower efficacy at GABA(A)-alpha3 receptors and least efficacy at GABA(A)-alpha1 and GABA(A)-alpha5 receptors. Because SL651498 has efficacy at GABA(A)-alpha1 receptors, we anticipated that it would generalize to both discriminative cues. Rats were trained to discriminate either zolpidem (3 mg/kg) or chlordiazepoxide (5 mg/kg) from vehicle using a two lever operant procedure. The generalization profiles of L-838,417 and SL651498 were compared with nonselective full agonists, GABA(A)-alpha1-selective ligands zolpidem and CL218,872 [3-methyl-6-[3-(trifluoromethyl)phenyl]-1,2,4-triazolo[4,3 b]pyridazine], the nonselective partial agonist bretazenil, and the novel anxioselective drug ocinaplon. A nonselective partial agonist was included because L-838,417 and SL651498 are partial agonists at some GABA(A) receptors, and this property may influence their generalization profiles. All nonselective full agonists and ocinaplon fully generalized to both cues. CL218,872 and zolpidem generalized to zolpidem only, whereas L-838,417 fully generalized to chlordiazepoxide only. SL651498 fully generalized to chlordiazepoxide and occasioned significant zolpidem-appropriate responding. Bretazenil was similar to SL651498. In conclusion, at this training dose, the chlordiazepoxide discriminative stimulus is mediated primarily via non-alpha1 GABA(A) receptors and the generalization profiles of the ligands tested seem to correspond with their in vitro profiles at GABA(A) receptor subtypes. PMID- 16339396 TI - Identification of a gene on chromosome 12q22 uniquely overexpressed in chronic lymphocytic leukemia. AB - The pathogenesis of chronic lymphocytic leukemia (CLL) is unknown but may involve aberrant activation of signaling pathways. Somatic hypermutations in rearranged immunoglobulin heavy-chain (IgVH) genes allow a division of CLL patients into 2 categories: mutated IgVH genes are associated with an indolent disease, whereas unmutated IgVH genes define an aggressive form. Using differential display to compare gene expression in CLL cells with and without IgVH hypermutations, we identified a novel gene, CLL up-regulated gene 1 (CLLU1), that was highly up regulated in CLL cells without IgVH hypermutations. CLLU1 mapped to chromosome 12q22, within a cluster of genes that are active in germinal center B cells. However, appreciable levels of CLLU1 were detectable only in CLL cells and not in a panel of normal tissue extracts or in any other tested hematologic malignancy. High expression of CLLU1 in CLL samples occurred irrespective of trisomy 12 or large chromosomal rearrangements. CLLU1 encodes 6 mRNAs with no sequence homology to any known gene, and most transcripts appear to be noncoding. Two transcripts, however, potentially encode a peptide with remarkable structural similarity to human interleukin 4. These data, in particular the unique and restricted expression pattern, suggest that CLLU1 is the first disease-specific gene identified in CLL. PMID- 16339397 TI - Lack of allosterically controlled intramolecular transfer of nitric oxide from the heme to cysteine in the beta subunit of hemoglobin. AB - The SNO-Hb hypothesis holds that heme-bound nitric oxide (NO) present in the beta subunits of T-state hemoglobin (Hb) will be transferred to the beta-93 cysteine upon conversion to R-state Hb, thereby forming SNO-Hb. A deficiency in the ability of Hb to facilitate this intramolecular transfer has recently been purported to play a role in pulmonary hypertension and sickle cell disease. We prepared deoxygenated Hb samples with small amounts of heme-bound NO and then oxygenated the samples. Electron paramagnetic resonance (EPR) spectroscopy was used to (1) determine the concentration of iron nitrosyl Hb (Fe-NO Hb), (2) show that the NO is evenly distributed among alpha and beta subunits, and (3) show that the Hb undergoes a change in its quaternary state (T to R) upon oxygenation. We did not observe a decrease in the concentration of Fe-NO Hb on oxygenation, which is inconsistent with the prediction of the SNO-Hb hypothesis. PMID- 16339398 TI - Chronic hepcidin induction causes hyposideremia and alters the pattern of cellular iron accumulation in hemochromatotic mice. AB - We report the generation of a tetracycline-regulated (Tet ON) transgenic mouse model for acute and chronic expression of the iron regulatory peptide hepcidin in the liver. We demonstrate that short-term and long-term tetracycline-dependent activation of hepcidin in adult mice leads to hypoferremia and iron-limited erythropoiesis, respectively. This clearly establishes the key role of hepcidin in regulating the extracellular iron concentration. We previously demonstrated that, when expressed early in fetal development, constitutive transgenic hepcidin expression prevented iron accumulation in an Hfe-/- mouse model of hemochromatosis. We now explore the effect of chronic hepcidin expression in adult Hfe-/- mice that have already developed liver iron overload. We demonstrate that induction of chronic hepcidin expression in 2-month-old Hfe-/- mice alters their pattern of cellular iron accumulation, leading to increased iron in tissue macrophages and duodenal cells but less iron in hepatocytes. These hepcidin induced changes in the pattern of cellular iron accumulation are associated with decreased expression of the iron exporter ferroportin in macrophages but no detectable alteration of ferroportin expression in the hepatocytes. We speculate that this change in iron homeostasis could offer a therapeutic advantage by protecting against damage to parenchymal cells. PMID- 16339399 TI - Flow cytometric detection of circulating myeloma cells before transplantation in patients with multiple myeloma: a simple risk stratification system. AB - Detection of circulating myeloma cells (CMCs) by flow cytometry in patients with multiple myeloma (MM) indicates active disease. We hypothesized that detection of CMCs at the time of stem-cell collection prior to autologous stem-cell transplantation (ASCT) identifies patients at high risk of rapid progression. A cohort of patients undergoing ASCT was identified. CMCs were determined by gating on CD38+/CD45- cells using flow cytometry. The impact of CMCs on overall survival (OS) and time to progression (TTP) was evaluated in univariate and multivariate analyses. Of 246 patients undergoing ASCT, 95 had CMCs. Complete response (CR) rates after transplantation were 32% and 36% for patients with and without CMCs, respectively (P = .50). OSs were 33.2 and 58.6 months (P = .01) whereas TTPs were 14.1 and 22 months, respectively (P = .001). On multivariate analysis, CMCs remained independent of cytogenetics and disease status at time of transplantation (P = .03). CMCs and cytogenetics were combined in a new scoring system. Patients with neither, one, or both parameters had a median OS of 55, 48, and 21.5 months and a median TTP of 22, 15.4, and 6.5 months, respectively. CMCs at the time of ASCT is an independent prognostic factor and in combination with cytogenetics provides a powerful scoring system that stratifies patients and guides management. PMID- 16339400 TI - Vaccination of human subjects expands both specific and bystander memory T cells but antibody production remains vaccine specific. AB - Human subjects maintain long-term immunologic memory against infective organisms but the mechanism is unclear. CD4+ T-helper memory (Thmem) cells are pivotal in controlling humoral and cellular responses, therefore their longevity and response to vaccination are critical for maintenance of protective immunity. To probe the dynamics of the Thmem-cell response to antigenic challenge, we investigated subjects following a booster injection with tetanus toxoid (TT). Expansion of TT-specific Thmem cells and cytokine production showed complex kinetics. Strikingly, parallel expansion and cytokine production occurred in pre existing Thmem cells specific for 2 other common antigens: purified protein derivative of tuberculin and Candida albicans. Bystander expansion occurred in Thmem but not in Thnaive cells. Antibody production against TT peaked approximately 2 weeks after vaccination and gradually declined. However, pre existing antibody against the other antigens did not change. It appears that although all Thmem cells are readily stimulated to expand, antibody responses are controlled by antigen availability. These findings relate to the maintenance of memory and have consequences for assessments of specific T-cell responses to vaccination. PMID- 16339401 TI - Toward the identification of a tolerogenic signature in IDO-competent dendritic cells. AB - Although much is known about the transcriptional profiles of dendritic cells (DCs) during maturation, the molecular switches critical for the induction of a tolerogenic program in DC subsets are still obscure. We examined the gene expression profiles of murine splenic CD8+ DCs rendered highly tolerogenic by interferon-gamma (IFN-gamma), which activates the enzyme indoleamine 2,3 dioxygenase (IDO, encoded by Indo) and thus initiates the immunosuppressive pathway of tryptophan catabolism. By examining the expression of a series of relevant genes in IDO+ compared with IDO- DCs, we found consistent and selective association of the IDO-competent phenotype with down-modulation of the Tyrobp gene, encoding the signaling adapter DAP12, which typically associates with activating receptors. Down-modulation of Tyrobp involved IFN consensus sequence binding protein (ICSBP), a transcription factor also known as IRF-8. In murine and human monocyte-derived DCs, silencing DAP12 expression imparted IDO functional competence to IDO- cells, whereas silencing IRF-8 in IDO+ counterparts abolished IDO expression and function. Thus, IRF-8 is required in tolerogenic DCs for the positive regulation of Indo and the negative regulation of Tyrobp. Overall, these studies reveal the occurrence of a simple and evolutionarily conserved code in the control of tolerance by an ancestral metabolic enzyme. PMID- 16339402 TI - Placental dermatan sulfate: isolation, anticoagulant activity, and association with heparin cofactor II. AB - Pregnancy is associated with hemostatic challenges that may lead to thrombosis. Heparin cofactor II (HCII) is a glycosaminoglycan-dependent thrombin inhibitor present in both maternal and fetal plasma. HCII activity increases during pregnancy, and HCII levels are significantly decreased in women with severe pre eclampsia. Dermatan sulfate (DS) specifically activates HCII and is abundant in the placenta, but the locations of DS and HCII in the placenta have not been determined. We present evidence that DS is the major anticoagulant glycosaminoglycan in the human placenta at term. DS isolated from human placenta contains disaccharides implicated in activation of HCII and has anticoagulant activity similar to that of mucosal DS. Immunohistochemical studies revealed that DS is associated with fetal blood vessels and stromal regions of placental villi but is notably absent from the syncytiotrophoblast cells in contact with the maternal circulation. HCII colocalizes with DS in the walls of fetal blood vessels and is also present in syncytiotrophoblast cells. Our data suggest that DS is in a position to activate HCII in the fetal blood vessels or in the stroma of placental villi after injury to the syncytiotrophoblast layer and thereby inhibit fibrin generation in the placenta. PMID- 16339403 TI - Lack of F8 mRNA: a novel mechanism leading to hemophilia A. AB - Hemophilia A (HA) is caused by partial or total deficiency of F8 protein activity. In a small group, about 1.8% of patients with HA, no mutation is found in the F8 gene. Among this group, we report here on one patient with severe HA in whom no mRNA of the F8 gene was detected. Using 2 common polymorphisms in F8 exon 14, we were able to show that the same allele shared by the patient, his mother, and his sister was not detected by reverse transcription-polymerase chain reaction (RT-PCR) from total blood mRNA. Skewed X-chromosome inactivation in both the mother and the sister was excluded by studying the methylation profile of the androgen receptor gene (HUMARA locus). These findings strongly suggest that the cause of HA in this patient is either absence or rapid degradation of the F8 mRNA, which points to a novel mechanism leading to HA. PMID- 16339404 TI - Long-term risk of cardiovascular disease after treatment for aggressive non Hodgkin lymphoma. AB - Cardiovascular disease frequently occurs after lymphoma therapy, but it is common in the general population too. Therefore, risk estimation requires comparison to population-based rates. We calculated risk by standardized incidence ratios (SIRs) and absolute excess risks (AERs) per 10,000 person-years based on general population rates (Continuous Morbidity Registry Nijmegen) in 476 (Dutch and Belgian) patients with aggressive non-Hodgkin lymphoma (NHL) treated with at least 6 cycles of doxorubicin-based chemotherapy in 4 European Organization for Research on Treatment of Cancer (EORTC) trials (1980-1999). Cumulative incidence of cardiovascular disease, estimated in a competing risk model, was 12% at 5 years and 22% at 10 years (median follow-up, 8.4 years). Risk of chronic heart failure appeared markedly increased (SIR, 5.4; 95% CI, 4.1-6.9) with an AER of 208 excess cases per 10 000 person-years, whereas risk of coronary artery disease matched the general population (SIR, 1.2; 95% CI, 0.8-1.8; AER, 8 per 10 000 person-years). Risk of stroke was raised (SIR, 1.8; 95% CI, 1.1-2.4; AER, 15 per 10 000 person-years), especially after additional radiotherapy (> 40 Gy). Preexisting hypertension, NHL at young age, and salvage treatment increased risk of all cardiovascular events; the effect of radiotherapy was dose dependent. In conclusion, patients are at long-term high risk of chronic heart failure after NHL treatment and need therefore life-long monitoring. In contrast, risk of coronary artery disease appeared more age dependent than treatment related. PMID- 16339405 TI - Evidence for incorporation of bone marrow-derived endothelial cells into perfused blood vessels in tumors. AB - Recent studies have demonstrated that the cellular contribution of the bone marrow to tumor neovascularization is highly complex. In this context, the extent to which bone marrow-derived cells incorporate as bona fide endothelial (nonhematopoietic) cells into perfused tumor vessels, or any new vessels formed postnatally (vasculogenesis), is unclear. To this end, we developed models to characterize local vessel-derived and bone marrow-derived endothelial cells (BMD ECs). Then, we characterized the BMD-ECs based on a set of endothelial markers and morphology. Finally, we quantified their contribution to perfused blood vessels in tumors using transplanted as well as spontaneous primary and metastatic tumor models. We demonstrate that BMD-ECs incorporate in perfused tumor vessels, and that this contribution varies with organ site and mouse strain. PMID- 16339406 TI - In vivo disruption of tolerogenic cross-presentation mechanisms uncovers an effective T-cell activation by B-cell lymphomas leading to antitumor immunity. AB - Bone marrow-derived antigen-presenting cells (APCs) play a central role in the induction of tolerance to tumor antigens expressed by B-cell lymphomas. Here we show that in vivo disruption of this APC-mediated tolerogenic mechanism unveils an intrinsic ability of malignant B cells to efficiently present tumor antigens to antigen-specific CD4+ T cells, resulting in a strong antitumor effect. This intrinsic antigen-presenting ability of malignant B cells is, however, overridden by tolerogenic bone marrow-derived APCs, leading instead to T-cell unresponsiveness and lack of antitumor effect. These results highlight the concept that therapeutic strategies aimed at enhancing the antigen-presenting function of B-cell lymphomas might not succeed unless the tolerogenic mechanisms mediated by bone marrow-derived APCs are disrupted in the first place. PMID- 16339407 TI - Analysis of HSC activity and compensatory Hox gene expression profile in Hoxb cluster mutant fetal liver cells. AB - Overexpression of Hoxb4 in bone marrow cells promotes expansion of hematopoietic stem cell (HSC) populations in vivo and in vitro, indicating that this homeoprotein can activate the genetic program that determines self-renewal. However, this function cannot be solely attributed to Hoxb4 because Hoxb4(-/-) mice are viable and have an apparently normal HSC number. Quantitative polymerase chain reaction analysis showed that Hoxb4(-/-) c-Kit+ fetal liver cells expressed moderately higher levels of several Hoxb cluster genes than control cells, raising the possibility that normal HSC activity in Hoxb4(-/-) mice is due to a compensatory up-regulation of other Hoxb genes. In this study, we investigated the competitive repopulation potential of HSCs lacking Hoxb4 alone, or in conjunction with 8 other Hoxb genes. Our results show that Hoxb4(-/-) and Hoxb1 b9 (-/-) fetal liver cells retain full competitive repopulation potential and the ability to regenerate all myeloid and lymphoid lineages. Quantitative Hox gene expression profiling in purified c-Kit+ Hoxb1-b9(-/-) fetal liver cells revealed an interaction between the Hoxa, b, and c clusters with variation in expression levels of Hoxa4,-a11, and -c4.Together, these studies show a complex network of genetic interactions between several Hox genes in primitive hematopoietic cells and demonstrate that HSCs lacking up to 30% of the active Hox genes remain fully competent. PMID- 16339408 TI - Expression of concern: Bombardier et al., "Comparison of upper gastrointestinal toxicity of rofecoxib and naproxen in patients with rheumatoid arthritis," N Engl J Med 2000;343:1520-8. PMID- 16339409 TI - Asymmetric hydrogenation of unfunctionalized, purely alkyl-substituted olefins. AB - Asymmetric hydrogenation of olefins is one of the most useful reactions for the synthesis of optically active compounds, especially in industry. However, the application range of the catalysts developed so far is limited to alkenes with a coordinating functional group or an aryl substituent next to the double bond. We have found a class of chiral iridium catalysts that give high enantioselectivity in the hydrogenation of unfunctionalized, trialkyl-substituted olefins. Because these catalysts do not require the presence of any particular functional group or aryl substituent in the substrate, they considerably broaden the scope of asymmetric hydrogenation. PMID- 16339410 TI - The distance to the Perseus spiral arm in the Milky Way. AB - We have measured the distance to the massive star-forming region W3OH in the Perseus spiral arm of the Milky Way to be 1.95 +/- 0.04 kiloparsecs (5.86 x10(16) km). This distance was determined by triangulation, with Earth's orbit as one segment of a triangle, using the Very Long Baseline Array. This resolves the long standing problem that there is a discrepancy of a factor of 2 between different techniques used to determine distances. The reason for the discrepancy is that this portion of the Perseus arm has anomalous motions. The orientation of the anomalous motion agrees with spiral density-wave theory, but the magnitude of the motion is somewhat larger than most models predict. PMID- 16339411 TI - Herpesviral protein networks and their interaction with the human proteome. AB - The comprehensive yeast two-hybrid analysis of intraviral protein interactions in two members of the herpesvirus family, Kaposi sarcoma-associated herpesvirus (KSHV) and varicella-zoster virus (VZV), revealed 123 and 173 interactions, respectively. Viral protein interaction networks resemble single, highly coupled modules, whereas cellular networks are organized in separate functional submodules. Predicted and experimentally verified interactions between KSHV and human proteins were used to connect the viral interactome into a prototypical human interactome and to simulate infection. The analysis of the combined system showed that the viral network adopts cellular network features and that protein networks of herpesviruses and possibly other intracellular pathogens have distinguishing topologies. PMID- 16339412 TI - Battle for the brains? PMID- 16339413 TI - Space science. NASA starts squeezing to fit missions into tight budget. PMID- 16339414 TI - Stem cells. Landmark paper has an image problem. PMID- 16339415 TI - Intellectual property. Cambridge University reins in faculty patents. PMID- 16339416 TI - Space science. Europe trumpets successes on Mars and Titan. PMID- 16339417 TI - Biomedical policy. U.K. doubles stem cell funding. PMID- 16339418 TI - European research. ERC moves forward despite budget impasse. PMID- 16339419 TI - NIH training grants. Universities may have to pay more in support of graduate training. PMID- 16339420 TI - NIH career awards. Young scientists get a helping hand. PMID- 16339421 TI - Indian Ocean tsunami. Girding for the next killer wave. PMID- 16339422 TI - Indian Ocean tsunami. A dead spot for the tsunami network? PMID- 16339423 TI - Indian Ocean tsunami. In the wake: looking for keys to posttraumatic stress. PMID- 16339424 TI - Infectious diseases. Will a preemptive strike against malaria pay off? PMID- 16339425 TI - Infectious diseases. Cracks in the first line of defense. PMID- 16339426 TI - Nanotechnology. Calls rise for more research on toxicology of nanomaterials. PMID- 16339427 TI - Energy. For nuclear fusion, could two lasers be better than one? PMID- 16339428 TI - Support for the Human Cancer Genome Project. PMID- 16339429 TI - Attribution of disaster losses. PMID- 16339430 TI - Bilateral action for right whales. PMID- 16339431 TI - Comment on "How science survived: medieval manuscripts' 'demography' and classic texts' extinction". AB - Exciting though it may seem, the mathematical model developed by Cisne (Reports, 25 February 2005, p. 1305) to analyze the transmission of texts and manuscripts from Antiquity and the Middle Ages does not hold up to scrutiny. It seriously underestimates the losses, thus leading to conclusions that are unwarranted. PMID- 16339432 TI - Agriculture. Losing the links between livestock and land. PMID- 16339433 TI - Materials science. Metallurgy in the age of silicon. PMID- 16339434 TI - Neuroscience. Emotion and reason in making decisions. PMID- 16339435 TI - Atmospheric science. Land use and climate change. PMID- 16339436 TI - Neuroscience. Synaptic membranes bend to the will of a neurotoxin. PMID- 16339437 TI - Restoration of degraded tropical forest landscapes. AB - The current scale of deforestation in tropical regions and the large areas of degraded lands now present underscore the urgent need for interventions to restore biodiversity, ecological functioning, and the supply of goods and ecological services previously used by poor rural communities. Traditional timber plantations have supplied some goods but have made only minor contributions to fulfilling most of these other objectives. New approaches to reforestation are now emerging, with potential for both overcoming forest degradation and addressing rural poverty. PMID- 16339438 TI - Increase in activity during calorie restriction requires Sirt1. AB - Sir2 (silent information regulator 2) is a nicotinamide adenine dinucleotide dependent deacetylase required for longevity due to calorie restriction in yeast and Drosophila. In mammals, calorie restriction induces a complex pattern of physiological and behavioral changes. Here we report that the mammalian Sir2 ortholog, Sirt1, is required for the induction of a phenotype by calorie restriction in mice. PMID- 16339439 TI - A systems model of signaling identifies a molecular basis set for cytokine induced apoptosis. AB - Signal transduction pathways control cellular responses to stimuli, but it is unclear how molecular information is processed as a network. We constructed a systems model of 7980 intracellular signaling events that directly links measurements to 1440 response outputs associated with apoptosis. The model accurately predicted multiple time-dependent apoptotic responses induced by a combination of the death-inducing cytokine tumor necrosis factor with the prosurvival factors epidermal growth factor and insulin. By capturing the role of unsuspected autocrine circuits activated by transforming growth factor-alpha and interleukin-1alpha, the model revealed new molecular mechanisms connecting signaling to apoptosis. The model derived two groupings of intracellular signals that constitute fundamental dimensions (molecular "basis axes") within the apoptotic signaling network. Projection along these axes captures the entire measured apoptotic network, suggesting that cell survival is determined by signaling through this canonical basis set. PMID- 16339440 TI - Rapid chiral assembly of rigid DNA building blocks for molecular nanofabrication. AB - Practical components for three-dimensional molecular nanofabrication must be simple to produce, stereopure, rigid, and adaptable. We report a family of DNA tetrahedra, less than 10 nanometers on a side, that can self-assemble in seconds with near-quantitative yield of one diastereomer. They can be connected by programmable DNA linkers. Their triangulated architecture confers structural stability; by compressing a DNA tetrahedron with an atomic force microscope, we have measured the axial compressibility of DNA and observed the buckling of the double helix under high loads. PMID- 16339441 TI - The chemistry of deformation: how solutes soften pure metals. AB - Solutes have been added to strengthen elemental metals, generating usable materials for millennia; in the 1960s, solutes were found to also soften metals. Despite the empirical correlation between the "electron number" of the solute and the change in strength of the material to which it is added, the mechanism responsible for softening is poorly understood. Using state-of-the-art quantum mechanical methods, we studied the direct interaction of transition-metal solutes with dislocations in molybdenum. The interaction increases dramatically with increasing electron number and strongly influences the mechanisms responsible for plasticity in these materials. Our quantitative model explains solution softening of metals by using changes in energy and stress scales of plasticity from solutes. PMID- 16339442 TI - Rapid glacial erosion at 1.8 Ma revealed by 4He/3He thermochronometry. AB - Alpine glaciation and river incision control the topography of mountain ranges, but their relative contributions have been debated for years. Apatite 4He/3He thermochronometry tightly constrains the timing and rate of glacial erosion within one of the largest valleys in the southern Coast Mountains of British Columbia, Canada. Five proximate samples require accelerated denudation of the Klinaklini Valley initiating 1.8 +/- 0.2 million years ago (Ma). At least 2 kilometers of overlying rock were removed from the valley at >/=5 millimeters per year, indicating that glacial valley deepening proceeded >/=6 times as fast as erosion rates before approximately 1.8 Ma. This intense erosion may be related to a global transition to enhanced climate instability approximately 1.9 Ma. PMID- 16339443 TI - The importance of land-cover change in simulating future climates. AB - Adding the effects of changes in land cover to the A2 and B1 transient climate simulations described in the Special Report on Emissions Scenarios (SRES) by the Intergovernmental Panel on Climate Change leads to significantly different regional climates in 2100 as compared with climates resulting from atmospheric SRES forcings alone. Agricultural expansion in the A2 scenario results in significant additional warming over the Amazon and cooling of the upper air column and nearby oceans. These and other influences on the Hadley and monsoon circulations affect extratropical climates. Agricultural expansion in the mid latitudes produces cooling and decreases in the mean daily temperature range over many areas. The A2 scenario results in more significant change, often of opposite sign, than does the B1 scenario. PMID- 16339444 TI - Equivalent effects of snake PLA2 neurotoxins and lysophospholipid-fatty acid mixtures. AB - Snake presynaptic phospholipase A2 neurotoxins (SPANs) paralyze the neuromuscular junction (NMJ). Upon intoxication, the NMJ enlarges and has a reduced content of synaptic vesicles, and primary neuronal cultures show synaptic swelling with surface exposure of the lumenal domain of the synaptic vesicle protein synaptotagmin I. Concomitantly, these neurotoxins induce exocytosis of neurotransmitters. We found that an equimolar mixture of lysophospholipids and fatty acids closely mimics all of the biological effects of SPANs. These results draw attention to the possible role of local lipid changes in synaptic vesicle release and provide new tools for the study of exocytosis. PMID- 16339445 TI - Neural systems responding to degrees of uncertainty in human decision-making. AB - Much is known about how people make decisions under varying levels of probability (risk). Less is known about the neural basis of decision-making when probabilities are uncertain because of missing information (ambiguity). In decision theory, ambiguity about probabilities should not affect choices. Using functional brain imaging, we show that the level of ambiguity in choices correlates positively with activation in the amygdala and orbitofrontal cortex, and negatively with a striatal system. Moreover, striatal activity correlates positively with expected reward. Neurological subjects with orbitofrontal lesions were insensitive to the level of ambiguity and risk in behavioral choices. These data suggest a general neural circuit responding to degrees of uncertainty, contrary to decision theory. PMID- 16339446 TI - A conserved checkpoint monitors meiotic chromosome synapsis in Caenorhabditis elegans. AB - We report the discovery of a checkpoint that monitors synapsis between homologous chromosomes to ensure accurate meiotic segregation. Oocytes containing unsynapsed chromosomes selectively undergo apoptosis even if a germline DNA damage checkpoint is inactivated. This culling mechanism is specifically activated by unsynapsed pairing centers, cis-acting chromosome sites that are also required to promote synapsis in Caenorhabditis elegans. Apoptosis due to synaptic failure also requires the C. elegans homolog of PCH2, a budding yeast pachytene checkpoint gene, which suggests that this surveillance mechanism is widely conserved. PMID- 16339447 TI - Snapshot of activated G proteins at the membrane: the Galphaq-GRK2-Gbetagamma complex. AB - G protein-coupled receptor kinase 2 (GRK2) plays a key role in the desensitization of G protein-coupled receptor signaling by phosphorylating activated heptahelical receptors and by sequestering heterotrimeric G proteins. We report the atomic structure of GRK2 in complex with Galphaq and Gbetagamma, in which the activated Galpha subunit of Gq is fully dissociated from Gbetagamma and dramatically reoriented from its position in the inactive Galphabetagamma heterotrimer. Galphaq forms an effector-like interaction with the GRK2 regulator of G protein signaling (RGS) homology domain that is distinct from and does not overlap with that used to bind RGS proteins such as RGS4. PMID- 16339448 TI - High seminal platelet-activating factor acetylhydrolase activity in men with spinal cord injury. AB - Spinal cord injury (SCI) causes male infertility, with low sperm motility the major long-term cause. It has been suggested in previous studies that some seminal components may be responsible for the pathological asthenozoospermia. It is hypothesized that platelet-activating factor (PAF) acetylhydrolase (PAFah), which originates in the epididymis and other accessory sexual glands, may be a causative factor. This enzyme catalyzes PAF to acetate and biologically inactive lyso-PAF. PAF is well recognized to be an important phospholipid mediator that stimulates sperm motility and enhances sperm capacitation and fertilization. The present study was designed to analyze differences in PAFah activity in semen of men with SCI and age-matched healthy men. PAFah assay reagent kits were used to measure enzymatic activity by monitoring the production rates of 4-nitrophenol on a spectrophotometer during a given interval. The results showed that subjects with SCI had a higher concentration of PAFah than men in the control group (P < .001). A statistically significant negative correlation was found between enzymatic activity and sperm motility (r(2) = 0.8449; P < .001). Further studies will determine whether seminal vesicle dysfunction in men with SCI leads to abnormal PAFah activity, resulting in low sperm motility. PMID- 16339449 TI - Role of oxidative stress in the pathophysiological mechanism of erectile dysfunction. PMID- 16339450 TI - Irradiated mouse testes efficiently support spermatogenesis derived from donor germ cells of mice and rats. AB - Testicular cell transplantation has been widely used to investigate the biology of spermatogonial stem cells, production of transgenic animals, and restoration of fertility in rodent models. One critical step in successful transplantation is the preparation of the recipient testes. Busulfan has been widely used, but irradiation has been often suggested as an alternative. There have only been limited reports of the use of irradiated animals as transplant recipients for studying differentiation of transplanted cells, and there has been no direct comparison of irradiation and busulfan as preparation methods. Mouse testes treated with local fractionated irradiation (1.5 + 12 Gy) were compared with busulfan-treated testes as recipients using mouse-to-mouse and rat-to-mouse germ cell transplantation. The fractionated irradiation schedule resulted in depletion of endogenous spermatogenesis similar to that produced by busulfan doses of 50-55 mg/kg. When immature mouse or rat testicular germ cells were transplanted into the irradiated testes, donor cells derived from either rat or mouse spermatogonial stem cells were able to form colonies of differentiated spermatogenic cells 10-13 weeks after transplantation with similar efficiencies as in busulfan-treated testes. Locally irradiated testes could be considered as an alternative to busulfan treatment for animal recipients of germ cell transplants that cannot endure the systemic toxicity of busulfan. PMID- 16339451 TI - Dissecting the protective effect of the seminal plasma spermadhesin PSP-I/PSP-II on boar sperm functionality. AB - To dissect the protective activity of PSP-I/PSP-II, the effect of the isolated subunits PSP-I and PSP-II and their affinity-purified tryptic peptide and glycan fractions on the viability, mitochondrial activity, and motility of highly diluted boar spermatozoa was investigated. High dilution exerted a negative effect on control spermatozoa. Incubation of spermatozoa with PSP-I/PSP-II or with its PSP-II subunit had a protective effect on sperm functionality, high mitochondrial membrane potential, and sperm motility. These effects were less pronounced when spermatozoa were incubated with the PSP-I subunit. It was noteworthy that motility was abolished by incubation of spermatozoa with isolated PSP-I. Trypsin-degraded PSP-I/PSP-II, PSP-I, and PSP-II reproduced the effects of the native proteins. Incubating spermatozoa with the glycan-depleted tryptic peptide fraction of PSP-I/PSP-II for 5 hours preserved a higher percentage of viable spermatozoa than when sperm was incubated for the same time with the native heterodimer, trypsin-digested PSP-I/PSP-II, the glycan fraction or without added proteins. However, sperm motility decreased as the concentration of added peptide fraction increased. On the other hand, spermatozoa incubated with the glycan fraction showed lower values than spermatozoa incubated with the peptide fraction. We concluded that the subunits of the PSP-I/PSP-II heterodimeric spermadhesin exert different activities on sperm functions. The finding that the beneficial effect of the native PSP-I/PSP-II on the functionality of highly diluted boar spermatozoa is largely preserved in its isolated PSP-II subunit and does not appear to require the glycan moiety points to a peptide moiety as a potential sperm function-preserving additive of highly diluted boar spermatozoa. PMID- 16339452 TI - Psychobiological correlates of delayed ejaculation in male patients with sexual dysfunctions. AB - The pathogenesis of delayed ejaculation (DE) is rather unknown, though the contribution of various psychological, marital, hormonal, and neurological factors has been advocated. In this study we systematically investigated the relative relevance of the aforementioned factors in 1,632 men who were seeking medical help for sexual dysfunction. The severity of DE was classified according to Kaplan criteria. Mild and moderate forms of DE (MMDE) recognized different risk factors than the most severe ones (anejaculation or severe DE [ASDE]). ASDE was essentially coupled with the presence of neurological diseases or with the use of serotoninergic drugs. Serotoninergic drugs also significantly increase (by at least 10-fold) the risk for MMDE, which, however, was also coupled with other relational factors (eg, partner's impaired climax, patient's hypoactive sexual desire [HSD]) or intrapsychic factors (eg, stress at work). At multiple regression analysis, some organic pathological conditions (such as psychiatric disorders and hypogonadism) were also associated with MMDE. In particular, hypogonadism retained significance for DE even after adjustment for HSD (adjusted odds ratio = 2.08 [1.11-3.89]; P < .05), suggesting other effects of testosterone deficiency on the ejaculatory reflex besides reduced libido. In conclusion, the present study demonstrates that multiple psychobiological determinants are associated with DE, a still obscure condition that substantially impairs psychosexual equilibrium of the couple. PMID- 16339453 TI - Role of metastin in the release of gonadotropin-releasing hormone from the hypothalamus of the male rat. AB - Recent genetic analysis has suggested that the expression of the orphan receptor GPR54 is essential for the onset of puberty in both rodents and humans. Indirect evidence has suggested that this action is via gonadotropin-releasing hormone induction of luteinizing hormone release. The experiments described here were intended to provide direct evidence that metastin, the naturally occurring ligand for GPR54, was capable of stimulating GnRH secretion by examining GnRH release from an immortalized hypothalamic cell line (GT1-7) and from male rat hypothalamic explants. GT1-7 cells were treated for 2(1/2) hours and overnight with the biologically active fragment of metastin, metastin(45-54), in amounts ranging from 0.1 nM to 1 muM. Hypothalamic fragments were obtained from infantile male rats and exposed to progressively increasing concentrations of metastin(45 54) (0.1 nM to 1 muM) for 1-hour periods. In both experiments, GnRH release was measured by radioimmunoassay (RIA). The release of metastin from hypothalami obtained from infantile and adult male rats was also determined. Explants were incubated for 6 hours, and the release of metastin into the media was determined by RIA. The results support the hypothesis that metastin stimulates GnRH secretion from the hypothalamus. The data indicate that an increase in the secretion of metastin, rather than the appearance of the receptor, is required for puberty onset. The results also suggest that metastin influences the GnRH secreting neurons indirectly via an interneuron rather than acting directly on the GnRH-secreting neurons. PMID- 16339454 TI - Impact of body mass index values on sperm quantity and quality. AB - Body mass index (BMI) has been demonstrated to affect female fertility; however, little information is available on the impact of BMI on male fertility or semen parameters. Therefore, the study objective was to determine the relationship between BMI and semen parameters, including sperm chromatin integrity. We analyzed data on semen samples from 520 men who were grouped based upon calculated BMI values (normal, 20-24 kg/m(2); overweight, 25-30 kg/m(2); obese, >30 kg/m(2)). The data collected included patient height and weight, semen volume, sperm concentration, percent sperm motility, percent sperm morphology (normal forms), and sperm chromatin integrity (DNA fragmentation index [DFI]). Data were analyzed by regression analysis and analysis of variance (ANOVA) with Tukey's test for multiple pairwise comparisons. The overall BMI mean (+/-SEM) was 27.5 (+/-0.49) kg/m(2). Linear regression revealed a significant (P < .05) and negative relationship between BMI and the total number of normal-motile sperm cells. ANOVA revealed a significant difference (P < .05) in the total number of normal-motile sperm cells among the different BMI groups. The number of normal motile sperm cells per BMI group was as follows: normal, 18.6 x 10(6); overweight, 3.6 x 10(6); and obese, (0.7) x 10(6). All multiple pairwise comparisons were found to be significantly (P < .05) different. The overall DFI mean (+/-SEM) was 24.7 (+/-2.57). Linear regression revealed a significant (P < .05) and positive relation between BMI and DFI. Men presenting with a BMI greater than 25 kg/m(2) have fewer chromatin-intact normal-motile sperm cells per ejaculate. Therefore, to ensure maximum fertility potential, patients may be advised to reduce body weight. PMID- 16339455 TI - Hyperactivated motility in rhesus macaque (Macaca mulatta) spermatozoa. AB - Macaque spermatozoa can be capacitated according to a defined protocol and exhibit hyperactivated motility similar to that described in other species. The aim of this study was to create a method for defining hyperactivation that could be routinely used in the laboratory alongside our existing sperm motility analysis protocol. Percoll-separated macaque spermatozoa were incubated for 2 hours (37 degrees C; 5% CO(2) in air) at a concentration of 20 x 10(6)/mL in bicarbonate (36 mmol)-buffered Biggers, Whitten and Whittingham medium (BWW) containing 30 mg/mL bovine serum albumin (BSA), followed by an additional 30 minutes with (capacitated) or without (incubated) caffeine (1 mmol) and dibutyryladenosine 3',5'-cyclic monophosphate (dbcAMP; 1.2 mmol). One hundred and fifty progressive and hyperactivated tracks were selected from each of three monkeys. Thresholds for hyperactivation were based on the 10th (amplitude of lateral head displacement, ALH) and 90th (linearity, LIN) percentiles of the hyperactivated kinematic data set and were LIN less than or equal to 69% and ALH greater than or equal to 7.5 microM; a threshold of greater than or equal to 130 microM/s was also included for curvilinear velocity (VCL). These thresholds were 91% effective at identifying hyperactivated tracks. Capacitation of macaque spermatozoa, by the addition of caffeine and dbcAMP, resulted in a significant increase in ALH, VCL, and beat cross frequency and a significant decrease in total and progressive motility, straight line velocity, straightness, and LIN when compared to incubated spermatozoa, suggesting the expression of hyperactivated motility. Utilizing the above thresholds, hyperactivation was expressed by 5% +/- 0.8% of the incubated sperm population vs 53 +/- 3.7% of the capacitated sperm population (P < .0001). Hyperactivation was not observed when dbcAMP and caffeine were added separately and was significantly (P < .005) reduced by the addition of H-89. The results of this paper demonstrate that hyperactivation can be reliably estimated for rhesus macaque spermatozoa. PMID- 16339456 TI - Factors affecting the increased prevalence of erectile dysfunction in Greek hypertensive compared with normotensive subjects. AB - Arterial hypertension is considered a risk factor for erectile dysfunction. The aim of the study was to evaluate the prevalence of erectile dysfunction in hypertensive compared with normotensive individuals of similar demographic characteristics in Greece. Furthermore, the effect of age, hypertension severity, hypertension duration, and antihypertension medication on erectile function of these subjects was investigated. The study population consisted of 634 consecutive young and middle-aged men (31-65 years) that visited our outpatient clinic. From them, 358 patients had arterial hypertension and 276 were normotensive. Erectile dysfunction was evaluated with the International Index for Erectile Function questionnaire. Erectile dysfunction was found in 35.2% of patients with essential hypertension compared with 14.1% of normotensive subjects (chi(2) = 35.92, P < .001). Patients with essential hypertension had more severe erectile dysfunction than their normotensive counterparts (chi(2) = 17.1, P < .001). Hypertension duration, hypertension severity, antihypertension medication, and age were positively correlated with erectile dysfunction. The prevalence of erectile dysfunction is higher in patients with essential hypertension compared with normotensive subjects of similar demographic characteristics. Erectile dysfunction is related to age in both groups, whereas duration and severity of hypertension as well as antihypertension drugs affect erectile function of hypertensive patients. Erectile dysfunction affects patient quality of life, underlining the need for vigorous research of this condition and appropriate management. PMID- 16339457 TI - The time man has cometh to brain: tick...death....tick...death. PMID- 16339459 TI - Intracerebral hemorrhage associated with oral anticoagulant therapy: current practices and unresolved questions. AB - BACKGROUND AND PURPOSE: Life-threatening intracranial hemorrhage, predominantly intracerebral hemorrhage (ICH), is the most serious complication of oral anticoagulant therapy (OAT), with mortality in excess of 50%. Early intervention focuses on rapid correction of coagulopathy in order to prevent continued bleeding. SUMMARY OF REVIEW: This article reviews the epidemiology of OAT associated ICH (OAT-ICH), and current treatment options, with the aim of providing a framework for future studies of unresolved questions. A number of acute treatments are available, but all have a significant risk of inducing thrombosis and other side effects, and vary in their rapidity of effect: vitamin K (very slow response time), fresh frozen plasma (slow response time, large volume of fluid required, transfusion-related acute lung injury), prothrombin complex concentrates, and recombinant activated factor VII. Current practice is to administer a combination of vitamin K and either fresh frozen plasma or prothrombin complex concentrates; the occasional use of recombinant activated factor VII has been reported. No prospective study has addressed the efficacy of, or outcomes from, the use of these practices. CONCLUSIONS: Current management of OAT-ICH is varied and not based on evidence from randomized controlled trials. Well-designed clinical trials are essential if we are to identify the effective acute treatments for OAT-ICH that are urgently needed. PMID- 16339458 TI - Long-term outcome after stroke: evaluating health-related quality of life using utility measurements. AB - BACKGROUND AND PURPOSE: As stroke mortality rates decline, individuals are increasingly likely to live with their residual impairments and disabilities. Therefore, the quality of poststroke life is 1 of the pivotal topics that have to be considered beneath the functional outcome. However, data on health-related quality of life (HRQoL) have been infrequently used in stroke trials. The purpose of this study was to examine the long-term outcome (4 years after stroke) of HRQoL and to identify the determinants of HRQoL in stroke survivors. METHODS: Seventy-seven patients were included who were admitted to the Department of Neurology, Philipps-University Marburg, after experiencing an ischemic stroke, a transient ischemic attack, or a hemorrhagic stroke. All patients were examined by a physician, and assessment was performed using a standardized questionnaire. HRQoL was assessed using the German version of the EuroQoL Index (EQ-5D) and the Health Utility Index 2 and 3 (HUI2/3). RESULTS: Four years after stroke, besides physical functioning, neuropsychological sequelae such as depression and cognitive impairment contributed to a reduced HRQoL. In addition, the incidence of incontinence proved to be an important factor for HRQoL. Explained variances in regression analysis models were high (R2=0.802 for HUI and 0.633 for EQ-5D- visual analogue scale) and were based on a few important determinants, including physical state, depression, cognitive impairment, and incontinence. CONCLUSIONS: Our results underscore the importance of nonmotor symptoms on HRQoL in patients with stroke. PMID- 16339460 TI - Telemedicine-guided carotid and transcranial ultrasound: a pilot feasibility study. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) and carotid duplex (CD) provide rapid and safe screening for stroke patients but are highly operator dependent. We explored the feasibility of telemedicine (TM)-guided TCD/CD administered by a health care provider inexperienced with ultrasound. METHODS: Dual video screens transmitted real-time TCD/CD images and sound to a neurosonographer. TM TCD/CD characteristics were compared with an in-person (IP) examination independently obtained on the same patient. We compared carotid stenosis, thrombolysis in brain ischemia (TIBI) flow grades, and the time spent on testing. RESULTS: We examined 8 subjects with a median age of 51 (31 to 63 range). IP and TM successfully examined 100% of internal carotid and middle cerebral arteries, 50% versus 44% of anterior cerebral artery, and 100% versus 88% of the basilar arteries, respectively. The median time in minutes IP versus TM was 15 (range 10 to 35) and 30 (15 to 50) for CD (P=0.07) and 18 (15 to 30) and 45 (30 to 55) for TCD (P=0.002), respectively. TM correctly identified all normal CD/TCD examinations in 7 subjects. In 1 patient, TM identified carotid occlusion but misread TIBI flow grades in both middle cerebral arteries. CONCLUSIONS: Our pilot study showed the feasibility of TCD/CD by an inexperienced health professional guided by a sonographer via TM. Tests were completed within times comparable to outpatient setting in a vascular laboratory. PMID- 16339461 TI - Matrix metalloproteinase 2 is associated with stable and matrix metalloproteinases 8 and 9 with vulnerable carotid atherosclerotic lesions: a study in human endarterectomy specimen pointing to a role for different extracellular matrix metalloproteinase inducer glycosylation forms. AB - BACKGROUND AND PURPOSE: We studied matrix metalloproteinases (MMP) 2, 8, and 9 and extracellular matrix metalloproteinase inducer (EMMPRIN) levels in relation to carotid atherosclerotic plaque characteristics. METHODS: Carotid atherosclerotic plaques (n=150) were stained and analyzed for the presence of collagen, smooth muscle cell (SMC), and macrophages. Adjacent segments were used to isolate total protein to assess MMP-2 and MMP-9 activities and gelatin breakdown, MMP-8 activity, and EMMPRIN levels. RESULTS: Macrophage-rich lesions revealed higher MMP-8 and MMP-9 activities, whereas SMC-rich lesions showed higher MMP-2 activity. The levels of less glycosylated EMMPRIN-45kD were higher in SMC-rich lesions and lower in macrophage-rich plaques. EMMPRIN-45kD was associated with MMP-2 levels, whereas EMMPRIN-58kD was related to MMP-9 levels. CONCLUSIONS: MMP-2, MMP-8, and MMP-9 activities differed among carotid plaque phenotypes. Different EMMPRIN glycosylation forms are associated with either MMP 2 or MMP-9 activity, which suggests that EMMPRIN glycosylation may play a role in MMP regulation and plaque destabilization. PMID- 16339462 TI - Validation of automatically classified magnetic resonance images for carotid plaque compositional analysis. AB - BACKGROUND AND PURPOSE: MRI may be used for noninvasive assessment of atherosclerotic lesions; however, MRI evaluation of plaque composition requires validation against an accepted reference standard, such as the American Heart Association (AHA) lesion grade, defined by histopathological examination. METHODS: Forty-eight carotid endarterectomy specimen cross-sections had AHA lesion grade determined histopathologically and were concurrently imaged using combinations of 8 MRI contrast weightings in vitro. A maximum likelihood classification algorithm generated MRI "maps" of plaque components, and an AHA lesion grade was assigned correspondingly. Additional analyses compared classification accuracy obtained with a commonly used set of magnetic resonance contrast weightings [proton density (PDw), T1 (T1w), and partial T2 (T2w)] to accuracy obtained with the combination of PDw, T1w, and diffusion-weighted (Dw) contrast. RESULTS: For the 8-contrast combination, the sensitivities for fibrous tissue, necrotic core, calcification, and hemorrhage detection were 83%, 67%, 86%, and 77%, respectively. The corresponding specificities were 81%, 78%, 99%, and 97%. Good agreement (79%) between magnetic resonance and histopathology for AHA classification was achieved. For the PDw, T1w, and Dw combination, the overall classification accuracy was insignificantly different at 78%, whereas the overall classification accuracy using PDw, T1w, and partial T2w contrast weightings was significantly lower at 67%. CONCLUSIONS: This study provides proof of-principle that the composition of atherosclerotic plaques determined by automated classification of high-resolution ex vivo MRI accurately reflects lesion composition defined by histopathological examination. PMID- 16339463 TI - Infratentorial abnormalities in vascular dementia. AB - BACKGROUND AND PURPOSE: Infratentorial abnormalities may cause cognitive deficits, but current research criteria for vascular dementia (VaD) do not consider them. Our purposes were to determine the prevalence of infratentorial abnormalities in VaD, their relation with supratentorial abnormalities, and whether they are relevant to cognition. METHODS: We examined 182 patients (120 men, mean age=73 years, SD=8) with probable VaD at inclusion into a multicenter clinical trial. MRI scans were evaluated for infratentorial vascular abnormalities, midbrain atrophy, cerebellar atrophy, basilar artery diameter and tortuosity, and supratentorial abnormalities. Cognitive testing included the mini mental state examination (MMSE) and the vascular dementia assessment scale (VaDAS cog). RESULTS: One hundred forty-one (77.5%) patients had infratentorial abnormalities: 119 (65.4%) had focal infratentorial vascular lesions, 65 (35.7%) had diffuse pontine vascular abnormalities hyperintense on T2-weighted images, 20 (11.0%) had midbrain atrophy, and 16 (8.8%) had cerebellar atrophy. Significant correlations were found between number of infratentorial vascular lesions and basilar artery diameter (rs=0.26; P<0.0001), infratentorial and basal ganglia (including thalamus) vascular abnormalities (rs=0.30; P<0.0001), as well as between midbrain atrophy and global supratentorial atrophy (rs=0.27; P<0.0001). Infratentorial vascular abnormalities and cerebellar atrophy were not significantly associated with cognitive impairment. Patients with midbrain atrophy performed worse on cognitive tests than those without midbrain atrophy. After correction for sex, age, education, supratentorial abnormalities, and center, midbrain atrophy remained significantly associated with lower MMSE scores (P<0.05). CONCLUSIONS: Infratentorial abnormalities often occur in patients with VaD, but only midbrain atrophy was found to be relevant to cognition. PMID- 16339464 TI - Cilostazol attenuates gray and white matter damage in a rodent model of focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: To evaluate whether delayed treatment with the antiplatelet agent cilostazol reduces the volume of infarction in the gray and white matter in a rodent model of permanent focal cerebral ischemia and to explore the mechanism of the neuroprotective effect in vivo. METHODS: Cilostazol (30 or 50 mg/kg) or vehicle was administered by gavage 30 minutes and 4 hours after the induction of cerebral ischemia by permanent occlusion of the left middle cerebral artery (MCA). Animals were euthanized 24 hours after MCA occlusion, and the volume of gray matter damage was evaluated by quantitative histopathology. Axonal damage was determined with amyloid precursor protein immunohistochemistry. Dynamic susceptibility contrast MRI was used to assess regional cerebral blood volume (CBV) and cerebral blood flow (CBF). RESULTS: Treatment with the higher dose of cilostazol (50 mg/kg) significantly reduced the volume of gray matter damage and axonal damage in the cerebral hemisphere by 45.0% (P<0.02) and 42.4% (P<0.002), respectively, compared with the control group. Relative CBV in the peri-infarct area after MCA occlusion was significantly increased in the cilostazol-treated group (50 mg/kg) compared with the control group (P<0.05). Relative CBF tended to be higher in the cilostazol treated group compared with the control group. CONCLUSIONS: Treatment with cilostazol significantly reduced the gray and white matter damage associated with permanent focal ischemia. Cilostazol improved CBV and CBF in the peri-infarct area. The major action of cilostazol is to increase perfusion in the ischemic penumbra. PMID- 16339465 TI - Carotid intima-media thickening indicates a higher vascular risk across a wide age range: prospective data from the Carotid Atherosclerosis Progression Study (CAPS). AB - BACKGROUND AND PURPOSE: Carotid intima-media thickness (IMT) is an independent predictor of vascular events in age groups >45 years. However, there is little information about the predictive value of IMT in younger individuals. METHODS: In the Carotid Atherosclerosis Progression Study (CAPS; n=5056; age range 19 to 90 years; mean age 50.1 years), common carotid artery (CCA) IMT, bifurcation IMT, internal carotid artery IMT and vascular risk factors were evaluated at baseline. The incidence of stroke, myocardial infarction (MI), and death was determined prospectively. Data for younger (<50 years; n=2436) and older subjects (> or =50 years; n=2620) were analyzed separately using Cox proportional hazard regression models. RESULTS: During a mean follow-up period of 4.2 years, there were 228 cases of MI, 107 strokes, and 50 deaths. IMT at all carotid segments was highly predictive of all end points (eg, hazard rate ratios [HRRs] per 1 SD CCA-IMT increase were 1.43 [95% CI: 1.35 to 1.51] for MI, 1.47 [1.35 to 1.60] for stroke, and 1.45 [1.38 to 1.52] for MI, stroke or death; all P<0.0001). Even after adjustment for age, sex, and vascular risk factors, the predictive value of CCA IMT and bifurcation IMT remained significant for MI and the combined end point. For the latter, the HRRs were considerably higher in the younger than in the older age group (eg, HRR per 0.1 mm CCA-IMT was 1.34 [1.16 to 1.55] vis-a-vis 1.10 [1.05 to 1.15]; P=0.011 for age-IMT interaction). CONCLUSIONS: Carotid IMT independently predicts future vascular events. Its predictive value is at least as high in younger subjects as in older subjects. PMID- 16339466 TI - Hyperperfusion syndrome after stenting for intracranial vertebral stenosis. AB - BACKGROUND AND PURPOSE: Hyperperfusion syndrome is a rare but well-described complication after endarterectomy or stenting in the carotid circulation. SUMMARY OF CASE: A 66-year-old man who had vertebrobasilar insufficiency refractory to medical treatment because of an intracranial right side vertebral stenosis was referred to our institution for endovascular treatment. Stenting was performed, and after 24 hours, he became extremely agitated, and this was followed by a period of apathy without focal neurological deficits. MRI showed bilateral thalamic hemorrhage. CONCLUSIONS: To our knowledge, this is the first report of hyperperfusion syndrome with hemorrhagic presentation after intracranial vertebral artery stenting. PMID- 16339467 TI - Time is brain--quantified. AB - BACKGROUND AND PURPOSE: The phrase "time is brain" emphasizes that human nervous tissue is rapidly lost as stroke progresses and emergent evaluation and therapy are required. Recent advances in quantitative neurostereology and stroke neuroimaging permit calculation of just how much brain is lost per unit time in acute ischemic stroke. METHODS: Systematic literature-review identified consensus estimates of number of neurons, synapses, and myelinated fibers in the human forebrain; volume of large vessel, supratentorial ischemic stroke; and interval from onset to completion of large vessel, supratentorial ischemic stroke. RESULTS: The typical final volume of large vessel, supratentorial ischemic stroke is 54 mL (varied in sensitivity analysis from 19 to 100 mL). The average duration of nonlacunar stroke evolution is 10 hours (range 6 to 18 hours), and the average number of neurons in the human forebrain is 22 billion. In patients experiencing a typical large vessel acute ischemic stroke, 120 million neurons, 830 billion synapses, and 714 km (447 miles) of myelinated fibers are lost each hour. In each minute, 1.9 million neurons, 14 billion synapses, and 12 km (7.5 miles) of myelinated fibers are destroyed. Compared with the normal rate of neuron loss in brain aging, the ischemic brain ages 3.6 years each hour without treatment. Altering single input variables in sensitivity analyses modestly affected the estimated point values but not order of magnitude. CONCLUSIONS: Quantitative estimates of the pace of neural circuitry loss in human ischemic stroke emphasize the time urgency of stroke care. The typical patient loses 1.9 million neurons each minute in which stroke is untreated. PMID- 16339468 TI - Midlife pulse pressure and incidence of dementia: the Honolulu-Asia Aging Study. AB - BACKGROUND AND PURPOSE: Previous studies have shown that midlife systolic blood pressure (SBP) predicts late-life cognitive decline and incident dementia. This study explores whether this association is attributable to the pulsatile, ie, pulse pressure (PP), or the nonpulsatile component of blood pressure (BP). METHODS: Data are from the Honolulu-Asia Aging Study, a community-based study of Japanese American men. Midlife BP was measured in 1971 to 1974 and dementia assessment was conducted in late-life. The 2505 men who were dementia free in 1991 and had complete follow-up data were re-examined for incident dementia in 1994 to 1996 and 1997 to 1999. Their age ranged from 71 to 93 years. Survival analysis with age as the time scale was performed to estimate the risk (hazard ratio [HR] and 95% CI) for incident dementia associated with mid- and late-life tertiles of PP and mean arterial BP, as well as SBP and diastolic BP categories. RESULTS: Over a mean of 5.1 years of follow-up, 189 cases (7.5%) of incident Alzheimer disease or vascular dementia were identified. After adjustment for cerebrovascular risk factors, dementia was significantly associated with SBP (HR 1.77; 95% CI, 1.10 to 2.84, for SBP > or =140 mm Hg compared with SBP <120 mm Hg), but not with PP tertiles. Limiting the analysis to those never treated with antihypertensives, high levels of all 4 BP components were significantly associated with dementia. In models with 2 BP components, only SBP remained significant in both the total sample and the never-treated subgroup (HR 2.29; 95% CI, 1.23 to 4.25, for SBP > or =140 mm Hg in total sample), whereas PP was not significantly associated with the risk for dementia. CONCLUSIONS: Midlife PP is not independently associated with dementia incidence. Midlife SBP is the strongest BP component predicting incident dementia. PMID- 16339469 TI - Task-specific training with trunk restraint on arm recovery in stroke: randomized control trial. AB - BACKGROUND AND PURPOSE: Task-specific training improves functional outcomes after stroke. However, gains may be accompanied by increases in movements compensating for motor impairments. We hypothesized that restriction of compensatory trunk movements may encourage recovery of premorbid movement patterns leading to better functional outcomes. The goal was to determine whether task-specific training with trunk-restraint (TR) produces greater improvements in arm impairment and function than training without TR in patients with chronic hemiparesis. METHODS: Double-blind randomized control trial of a therapist-supervised home program (3 times per week, 5 weeks) in 30 patients with chronic hemiparesis stratified by arm impairment level (Fugl-Meyer) was performed. Intervention group (TR group) received progressive object-related reach-to-grasp training with prevention of trunk movements. Control group (C) practiced tasks without TR. Main outcome measures were upper limb impairment (Fugl-Meyer Arm Section) and function (TEMPA) and movement kinematics (trunk displacement, elbow extension; Optotrak, 10 trials) of a reach-to-grasp movement. Evaluations were repeated before, immediately after, and 1 month postintervention by blind evaluators. RESULTS: TR training led to greater improvements in impairment and function compared with C. Improvements were accompanied by increased active joint range and were greater in initially more severe patients. In these patients, TR decreased trunk movement and increased elbow extension, whereas C had opposite effects (increased compensatory movements). In TR, changes in arm function were correlated with changes in arm and trunk kinematics. CONCLUSIONS: Treatment should be tailored to arm impairment severity with particular attention to controlling excessive trunk movements if the goal is to improve arm movement quality and function. PMID- 16339470 TI - Predictors of long-term mortality in patients with ischemic stroke referred for transesophageal echocardiography. AB - BACKGROUND AND PURPOSE: Findings on transesophageal echocardiography (TEE) after ischemic stroke predict recurrent embolic events and prompt therapy; however, the additive predictive power of TEE findings on long-term mortality is unknown. Our goal was to study the impact of TEE findings on all cause mortality in ischemic stroke patients referred for TEE. METHODS: We reviewed 245 consecutive patients who underwent TEE for ischemic stroke of undetermined origin (2000 to 2003). Long term survival was assessed using the Social Security Death Index. RESULTS: In a mean follow-up period of 3.0 (1.4 to 4.8) years, death occurred in 19.2% of patients. TEE findings included patent foramen ovale (18.8%), left atrium/left ventricle thrombus (2.4%), spontaneous echo contrast (3.7%), atrial septal aneurysm (3.3%), valve vegetation/mass/tumor (7.8%), complex aortic atheroma ([CAA]; 14.7%), and the composite of any cardiac source of embolus (39.2%). A total atherosclerotic burden (TAB) score was also recorded. On Cox hazard regression analysis, measures of aortic atherosclerosis (CAA [hazard ratio (HR), 2.7; 95% CI, 1.4 to 5.3] or TAB score [HR, 1.4; 95% CI, 1.2 to 1.6]) were independent predictors of death, whereas other TEE findings were not. CONCLUSIONS: In patients with ischemic stroke of undetermined origin referred for TEE, measures of aortic atherosclerosis, including CAA, represent the only TEE findings that predict long-term mortality after all other clinical factors are considered. Further study is needed to determine whether treatments for CAA effect long-term survival in patients with ischemic stroke. PMID- 16339472 TI - Does hyperglycemia contribute to secondary injury in subarachnoid hemorrhage? PMID- 16339474 TI - Genetics of vascular cognitive impairment: the opportunity and the challenges. AB - BACKGROUND AND PURPOSE: This review considers the current state of knowledge of genetic factors underlying vascular cognitive impairment (VCI). SUMMARY OF REVIEW: We argue here that genes conferring susceptibility to VCI must be of 2 nonmutually exclusive classes: (1) genes that confer susceptibility to cerebrovascular disease, and (2) genes that determine brain tissue responses to cerebrovascular disease (ie, render parenchymal tissue more or less susceptible to injury or able to repair itself after injury). Although some progress has been made in identifying genes of the first class, little has been done to explore genes of the second class. Evidence for the existence of such genes is presented. We discuss the advantages and disadvantages of different forms of cerebrovascular disease for studying these genes, and different study designs that might be used. CONCLUSIONS: The most critical challenge for genetic studies of VCI is to identify quantifiable phenotypes that can be reliably and effectively determined in large samples of subjects. PMID- 16339473 TI - Mechanical thrombolysis in ischemic stroke attributable to basilar artery occlusion as first-line treatment. AB - BACKGROUND AND PURPOSE: To report results of mechanical disruption or retrieval of thrombus as first-line treatment in patients with stroke attributable to occlusion of the basilar artery, in particular regarding efficiency and safety. METHODS: In 12 consecutive patients with acute stroke attributable to basilar occlusion, mechanical disruption or thrombus retrieval using various loop-shaped tools was tried before eventually starting local intra-arterial thrombolysis with recombinant tissue plasminogen activator (r-tPA). Main inclusion criteria were: National Institutes of Health Stroke Scale score >8 or Glasgow Coma Scale score <12; onset or worsening of symptoms <8 hours; no hemorrhages or large hypodensities on computed tomography scan; and occlusion of the basilar artery matching clinical symptoms. Efficiency included recanalization, procedure time, and r-tPA dose; safety was defined as rate of procedure-related complications. Outcome was evaluated at 3 months. RESULTS: Mechanical recanalization was successful in 6 patients. A single brain infarction, possibly attributable to distal embolization, occurred. Three patients had good outcomes. In 5 of 6 remaining patients, the artery was recanalized using r-tPA. A single asymptomatic hemorrhage occurred; 3 patients had good outcomes. Procedure time and r-tPA were significantly less in patients with successful mechanical thrombolysis (43.33 minutes and 13.33 mg versus 112.33 minutes and 55.83 mg, respectively). CONCLUSIONS: Mechanical recanalization was effective in half of the patients and at least as safe as local intra-arterial thrombolysis. It allowed to save r-tPA and time. Although the low success rate remains a limit, the excellent and quick anatomical recanalization obtained after successful procedures makes this approach promising. PMID- 16339475 TI - Prolonged low-dose intravenous thrombolysis in a stroke patient with distal basilar thrombus. AB - BACKGROUND AND PURPOSE: Patients with high-grade basilar artery stenosis secondary to thromboembolism are at high risk of developing subsequent vessel occlusion. Optimal medical management of this condition is unclear. SUMMARY OF CASE: We present a patient with a small subacute brain stem infarction and filiform distal basilar residual lumen attributable to arterioarterial or cardiogenic embolism. Beginning 3 days after symptom onset, low-dose intravenous thrombolysis with 0.125 mg/kg recombinant tissue plasminogen activator was continuously infused for 48 hours. Follow-up magnetic resonance angiography revealed complete resolution of the embolus. No further cerebral ischemic episodes occurred during 3-month follow-up, and the basilar artery remained patent. CONCLUSIONS: Our observation suggests a potential for prolonged low-dose intravenous thrombolysis in basilar artery embolism, but further data are needed to judge the effectiveness and risk of this intervention. PMID- 16339476 TI - Dietary intake of calcium in relation to mortality from cardiovascular disease: the JACC Study. AB - BACKGROUND AND PURPOSE: No prospective studies have examined the association between calcium intake and the risk of cardiovascular disease in Japanese populations with a low mean calcium intake. METHODS: Between 1988 and 1990, 110,792 Japanese subjects (46,465 men and 64,327 women) 40 to 79 years of age without a history of stroke, coronary heart disease, or cancer, completed a lifestyle questionnaire including food intake frequency under the Japan Collaborative Cohort (JACC) Study for Evaluation of Cancer Risk Sponsored by Monbusho. By the end of 1999, after 515,029 person years of follow-up, 566 deaths from stroke (101 subarachnoid hemorrhages, 140 intraparenchymal hemorrhages, and 273 ischemic strokes) and 234 deaths from coronary heart disease had been documented. RESULTS: The intake of total calcium tended to be inversely associated with mortality from total stroke but not from coronary heart disease or total cardiovascular disease for men and women. The inverse association with dairy calcium intake was apparent for total stroke, both hemorrhagic and ischemic. The multivariate relative risk for men with highest versus lowest quintiles of dairy calcium intake was 0.53 (95% CI, 0.34 to 0.81) for total stroke, 0.46 (0.23 to 0.91) for hemorrhagic stroke, and 0.53 (0.29 to 0.99) for ischemic stroke; corresponding relative risks for women were 0.57 (0.38 to 0.86), 0.51 (0.28 to 0.94), and 0.50 (0.27 to 0.95). CONCLUSIONS: Dietary calcium intake from dairy products was associated with reduced mortality from stroke for Japanese men and women. PMID- 16339477 TI - Trends in ethnic disparities in stroke incidence in Auckland, New Zealand, during 1981 to 2003. AB - BACKGROUND AND PURPOSE: Although geographical variations in stroke rates are well documented, limited data exist on temporal trends in ethnic-specific stroke incidence. METHODS: We assessed trends in ethnic-specific stroke rates using standard diagnostic criteria and community-wide surveillance procedures in Auckland, New Zealand (NZ) in 1981 to 1982, 1991 to 1992, and 2002 to 2003. Indirect and direct methods were used to adjust first-ever (incident) and total (attack) rates for changes in the structure of the population and reported with 95% CIs. Ethnicity was self-defined and categorized as "NZ/European," "Maori," "Pacific peoples," and "Asian and other." RESULTS: Stroke attack (19%; 95% CI, 11% to 26%) and incidence rates (19%; 95% CI, 12% to 24%) declined significantly in NZ/Europeans from 1981 to 1982 to 2002 to 2003. These rates remained high or increased in other ethnic groups, particularly for Pacific peoples in whom stroke attack rates increased by 66% (95% CI; 11% to 225%) over the periods. Some favorable downward trends in vascular risk factors, such as cigarette smoking, were counterbalanced by increasing age, body mass index, and diabetes in certain ethnic groups. CONCLUSIONS: Divergent trends in ethnic-specific stroke incidence and attack rates, and of associated risk factors, have occurred in Auckland over recent decades. The findings provide mixed views as to the future burden of stroke in populations undergoing similar lifestyle and structural changes. PMID- 16339480 TI - Potential consequences for recruitment, power, and external validity of requirements for additional risk factors for eligibility in randomized controlled trials in secondary prevention of stroke. AB - BACKGROUND AND PURPOSE: Eligibility criteria determine the external validity (generalizability) of the results of randomized controlled trials. To increase the number of outcome events, and hence statistical power, some recent stroke prevention trials have required additional vascular risk factors for eligibility. METHODS: To assess the merits of additional eligibility criteria in stroke prevention trials, we analyzed data from 3 trials and 1 hospital-referred series of patients with a transient ischemic attack or minor ischemic stroke. Patients were stratified according to 2 sets of additional risk factors similar to those used in recent trials (MATCH, SPORTIF and PRoFESS); risk of stroke, myocardial infarction, or vascular death was calculated in relation to the number of risk factors. RESULTS: Although the observed risk during follow-up did increase with the number of risk factors present (P<0.01 for both sets), the risks in patients with > or =1 risk factors were not substantially greater than those in all patients. Consequently, although the proportions of patients with no risk factors in the 4 cohorts differed substantially between the 2 sets of eligibility criteria (21% to 28% versus 56% to 73%), in neither case could their exclusion be justified on statistical grounds. CONCLUSIONS: The degree of patient selection introduced by use of additional vascular risk factors as eligibility criteria for trials can differ substantially between apparently similar sets of risk factors. Given that the potential for additional eligibility criteria to undermine generalizability and prolong recruitment outweighs any benefits in terms of statistical power, the exclusion of patients with no risk factors is difficult to justify. PMID- 16339479 TI - Lipid testing and lipid-lowering therapy in hospitalized ischemic stroke and transient ischemic attack patients: results from a statewide stroke registry. AB - BACKGROUND AND PURPOSE: Recent recommendations call for in-hospital initiation of lipid-lowering therapy (LLT) for most ischemic stroke (IS) and transient ischemic attack (TIA) survivors; however, little is known about actual use. This study describes use of and predictors for in-hospital lipid testing and LLT using data from a statewide stroke registry. METHODS: In 2002, the registry ascertained cases from a stratified sample of 16 hospitals. This study includes only IS and TIA cases discharged alive. RESULTS: In 1907 study subjects, 30.2% (27.2% to 33.5%) were on LLT at admission. In 1399 subjects not on LLT at admission, 37.2% (30.2% to 44.9%) underwent lipid testing, and 12.9% (7.2% to 22.1%) received LLT at discharge. Use of testing and LLT varied widely between hospitals (P<0.001). In-hospital lipid testing was positively associated with large teaching hospitals (P=0.029), and neurologist or neurosurgeon (P=0.004); and negatively associated with increasing age (P=0.002), being female (P=0.020), a previous medical history of atrial fibrillation (P=0.002), nonambulatory status (P=0.005), and poor prognosis (P<0.001). LLT at discharge was positively associated with a previous medical history of dyslipidemia (P<0.001), lipid testing (P=0.004), and elevated low-density lipoprotein levels (P<0.001). Among subjects who were not on LLT at admission but who had Adult Treatment Panel III-based indications for use of LLT, only 31.2% (20.5% to 44.5%) received LLT at discharge. CONCLUSIONS: Many hospitalized acute IS and TIA patients with indications for LLT are untreated at discharge. Efforts to close treatment gaps in lipid evaluation and treatment require sustained quality improvement efforts and should pay particular attention to high-risk patients. PMID- 16339481 TI - Hyperglycemia after SAH: predictors, associated complications, and impact on outcome. AB - BACKGROUND AND PURPOSE: Hyperglycemia is common after subarachnoid hemorrhage (SAH). The extent to which prolonged hyperglycemia contributes to in-hospital complications and poor outcome after SAH is unknown. METHODS: We studied an inception cohort of 281 SAH patients with an initial serum glucose level obtained within 3 days of SAH onset and who had at least 7 daily glucose measurements between SAH days 0 and 10. We defined mean glucose burden (GB) as the average peak daily glucose level >5.8 mmol/L (105 mg/dL). Hospital complications were recorded prospectively, and 3-month outcome was assessed with the modified Rankin scale. RESULTS: The median GB was 1.8 mmol/L (33 mg/dL). Predictors of high-GB included age > or =54 years, Hunt and Hess grade III-V, poor Acute Physiology and Chronic Health Evaluation (APACHE)-2 physiological subscores, and a history of diabetes mellitus (all P< or =0.001). In a multivariate analysis, GB was associated with increased intensive care unit length of stay (P=0.003) and the following complications: congestive heart failure, respiratory failure, pneumonia, and brain stem compression from herniation (all P<0.05). After adjusting for Hunt-Hess grade, aneurysm size, and age, GB was an independent predictor of death (odds ratio, 1.10 per mmol/L; 95% CI, 1.01 to 1.21; P=0.027) and death or severe disability (modified Rankin scale score of 4 to 6; odds ratio, 1.17 per mmol/L; 95% CI 1.07 to 1.28, P<0.001). CONCLUSIONS: Hyperglycemia after SAH is associated with serious hospital complications, increased intensive care unit length of stay, and an increased risk of death or severe disability. PMID- 16339482 TI - Evidence of structural remodeling in the dyssynchronous failing heart. AB - Ventricular remodeling of both geometry and fiber structure is a prominent feature of several cardiac pathologies. Advances in MRI and analytical methods now make it possible to measure changes of cardiac geometry, fiber, and sheet orientation at high spatial resolution. In this report, we use diffusion tensor imaging to measure the geometry, fiber, and sheet architecture of eight normal and five dyssynchronous failing canine hearts, which were explanted and fixed in an unloaded state. We apply novel computational methods to identify statistically significant changes of cardiac anatomic structure in the failing and control heart populations. The results demonstrate significant regional differences in geometric remodeling in the dyssynchronous failing heart versus control. Ventricular chamber dilatation and reduction in wall thickness in septal and some posterior and anterior regions are observed. Primary fiber orientation showed no significant change. However, this result coupled with the local wall thinning in the septum implies an altered transmural fiber gradient. Further, we observe that orientation of laminar sheets become more vertical in the early-activated septum, with no significant change of sheet orientation in the late-activated lateral wall. Measured changes in both fiber gradient and sheet structure will affect both the heterogeneity of passive myocardial properties as well as electrical activation of the ventricles. PMID- 16339483 TI - Type I collagen limits VEGFR-2 signaling by a SHP2 protein-tyrosine phosphatase dependent mechanism 1. AB - During angiogenesis, a combined action between newly secreted extracellular matrix proteins and the repertoire of integrins expressed by endothelial cells contributes in the regulation of their biological functions. Extracellular matrix engaged integrins influence tyrosine kinase receptors, thus promoting a regulatory cross-talk between adhesive and soluble stimuli. For instance, vitronectin has been reported to positively regulate VEGFR-2. Here, we show that collagen I downregulates VEGF-A-mediated VEGFR-2 activation. This activity requires the tyrosine phosphatase SHP2, which is recruited to the activated VEGFR 2 when cells are plated on collagen I, but not on vitronectin. Constitutive expression of SHP2(C459S) mutant inhibits the negative role of collagen I on VEGFR-2 phosphorylation. VEGFR-2 undergoes internalisation, which is associated with dynamin II phosphorylation. Expression of SHP2(C459S) impairs receptor internalisation suggesting that SHP2-dependent dephosphorylation regulates this process. These findings demonstrate that collagen I in provisional extracellular matrix surrounding nascent capillaries triggers a signaling pathway that negatively regulates angiogenesis. PMID- 16339484 TI - Cardiac myocyte apoptosis is associated with increased DNA damage and decreased survival in murine models of obesity. AB - Disruption of leptin signaling is associated with obesity, heart failure, and cardiac hypertrophy, but the role of leptin in cardiac myocyte apoptosis is unknown. We tested the hypothesis that apoptosis increases in leptin-deficient ob/ob and leptin-resistant db/db mice and is associated with aging and left ventricular hypertrophy, increased DNA damage, and decreased survival. We studied young (2- to 3-month-old) and old (12- to 14-month-old) ob/ob and db/db mice and wild-type (WT) controls (n=2 to 4 per group). As expected, ventricular wall thickness and heart weights were similar among young ob/ob, db/db, and WT mice, but higher in old ob/ob and db/db versus old WT. Young ob/ob and db/db showed markedly elevated apoptosis by TUNEL staining and caspase 3 levels compared with WT. Differences in apoptosis were further accentuated with age. Leptin treatment significantly reduced apoptosis in ob/ob mice both in intact hearts and isolated myocytes. Tissue triglycerides were increased in ob/ob hearts, returning to WT levels after leptin repletion. Furthermore, the DNA damage marker, 8oxoG (8-oxo 7,8-dihydroguanidine), was increased, whereas the DNA repair marker, MYH glycosylase, was decreased in old ob/ob and db/db compared with old WT mice. Both ob/ob and db/db mice had decreased survival compared with WT mice. We conclude that leptin-deficient and leptin-resistant mice demonstrate increased apoptosis, DNA damage, and mortality compared with WT mice, suggesting that normal leptin signaling is necessary to prevent excess age-associated DNA damage and premature mortality. These data offer novel insights into potential mechanisms of myocardial dysfunction and early mortality in obesity. PMID- 16339485 TI - Arrhythmogenic mutation-linked defects in ryanodine receptor autoregulation reveal a novel mechanism of Ca2+ release channel dysfunction. AB - Arrhythmogenic cardiac ryanodine receptor (RyR2) mutations are associated with stress-induced malignant tachycardia, frequently leading to sudden cardiac death (SCD). The causative mechanisms of RyR2 Ca2+ release dysregulation are complex and remain controversial. We investigated the functional impact of clinically severe RyR2 mutations occurring in the central domain, and the C-terminal I domain, a key locus of RyR2 autoregulation, on interdomain interactions and Ca2+ release in living cells. Using high-resolution confocal microscopy and fluorescence resonance energy transfer (FRET) analysis of interaction between fusion proteins corresponding to amino- (N-) and carboxyl- (C-) terminal RyR2 domains, we determined that in resting cells, RyR2 interdomain interaction remained unaltered after introduction of SCD-linked mutations and normal Ca2+ regulation was maintained. In contrast, after channel activation, the abnormal Ca2+ release via mutant RyR2 was intrinsically linked to altered interdomain interaction that was equivalent with all mutations and exhibited threshold characteristics (caffeine >2.5 mmol/L; Ca2+ >150 nmol/L). Noise analysis revealed that I domain mutations introduced a distinct pattern of conformational instability in Ca2+ handling and interdomain interaction after channel activation that was absent in signals obtained from the central domain mutation. I domain linked channel instability also occurred in intact RyR2 expressed in CHO cells and in HL-1 cardiomyocytes. These new insights highlight a critical role for mutation-linked defects in channel autoregulation, and may contribute to a molecular explanation for the augmented Ca2+ release following RyR2 channel activation. Our findings also suggest that the mutational locus may be an important mechanistic determinant of Ca2+ release channel dysfunction in arrhythmia and SCD. PMID- 16339486 TI - Raised plasma soluble P-selectin in peripheral arterial occlusive disease enhances leukocyte adhesion. AB - Raised levels of soluble P-selectin (sP-selectin) have been reported in the plasma of patients with vascular diseases; however, the functional importance of this ligand remains unclear. In this study we have examined a potential role for plasma sP-selectin in regulating neutrophil adhesion in patients with peripheral arterial occlusive disease (PAOD). Patients with PAOD had significantly higher levels of sP-selectin (mean+/-SD: 73.3+/-13.0 versus 16.7+/-6.4 ng/mL) and enhanced whole blood leukocyte adhesion to platelets under shear. To examine whether the raised sP-selectin levels can directly influence leukocyte adhesion, isolated neutrophils were incubated with plasma from PAOD patients before and after immunodepletion of sP-selectin. Neutrophil adhesion to fibrinogen increased 2-fold following incubation with PAOD plasma, which was abrogated on sP-selectin immunodepletion. We subsequently demonstrated that recombinant sP-selectin dose dependently (75 to 250 ng/mL) increased leukocyte adhesion to fibrinogen and platelet monolayers. This increase was PSGL-1 and Src kinase-dependent and correlated with an increase in sP-selectin-mediated Mac-1 activation. sP-selectin stimulated neutrophil adhesion to platelet monolayers was inversely correlated with shear, such that at low shear (50 s(-1)) a 92.7%+/-15.7 increase in adhesion was observed decreasing to 38.5%+/-11.9 at 150 s(-1) and 10.1%+/-7.4 at 300 s( 1). These studies suggest a potentially important role for sP-selectin in modulating neutrophil adhesion in patients with PAOD, particularly at sites of low shear, where it raises the possibility that raised plasma sP-selectin levels may enhance leukocyte recruitment to vascular injury and promote disease progression. PMID- 16339487 TI - High-density lipoprotein promotes endothelial cell migration and reendothelialization via scavenger receptor-B type I. AB - Vascular disease risk is inversely related to circulating levels of high-density lipoprotein (HDL) cholesterol. However, the mechanisms by which HDL provides vascular protection are unclear. The disruption of endothelial monolayer integrity is an important contributing factor in multiple vascular disorders, and vascular lesion severity is tempered by enhanced endothelial repair. Here, we show that HDL stimulates endothelial cell migration in vitro in a nitric oxide independent manner via scavenger receptor B type I (SR-BI)-mediated activation of Rac GTPase. This process does not require HDL cargo molecules, and it is dependent on the activation of Src kinases, phosphatidylinositol 3-kinase, and p44/42 mitogen-activated protein kinases. Rapid initial stimulation of lamellipodia formation by HDL via SR-BI, Src kinases, and Rac is also demonstrable. Paralleling the in vitro findings, carotid artery reendothelialization after perivascular electric injury is blunted in apolipoprotein A-I(-/-) mice, and reconstitution of apolipoprotein A-I expression rescues normal reendothelialization. Furthermore, reendothelialization is impaired in SR-BI(-/-) mice. Thus, HDL stimulates endothelial cell migration via SR-BI-initiated signaling, and these mechanisms promote endothelial monolayer integrity in vivo. PMID- 16339488 TI - Differential regulation of hyaluronic acid synthase isoforms in human saphenous vein smooth muscle cells: possible implications for vein graft stenosis. AB - Autologous saphenous vein bypass grafts (SVG) are frequently compromised by neointimal thickening and subsequent atherosclerosis eventually leading to graft failure. Hyaluronic acid (HA) generated by smooth muscle cells (SMC) is thought to augment the progression of atherosclerosis. The aim of the present study was (1) to investigate HA accumulation in native and explanted arterialized SVG, (2) to identify factors that regulate HA synthase (HAS) expression and HA synthesis, and (3) to study the function of the HAS2 isoform. In native SVG, expression of all 3 HAS isoforms was detected by RT-PCR. Histochemistry revealed that native and arterialized human saphenous vein segments were characterized by marked deposition of HA in association with SMC. Interestingly, in contrast to native SVG, cyclooxygenase (COX)-2 expression by SMC and macrophages was detected only in arterialized SVG. In vitro in human venous SMC HAS isoforms were found to be differentially regulated. HAS2, HAS1, and HA synthesis were strongly induced by vasodilatory prostaglandins via Gs-coupled prostaglandin receptors. In addition, thrombin induced HAS2 via activation of PAR1 and interleukin 1beta was the only factor that induced HAS3. By small interfering RNA against HAS2, it was shown that HAS2 mediated HA synthesis is critically involved in cell cycle progression through G1/S phase and SMC proliferation. In conclusion, the present study shows that HA-rich extracellular matrix is maintained after arterialization of vein grafts and might contribute to graft failure because of its proproliferative function in venous SMC. Furthermore, COX-2-dependent prostaglandins may play a key role in the regulation of HA synthesis in arterialized vein grafts. PMID- 16339489 TI - In vivo effects of C-reactive protein (CRP)-infusion into humans. PMID- 16339490 TI - EDHF redux: EETs, TRPV4, and Ca2+ sparks. PMID- 16339491 TI - Potassium channels and proliferation of vascular smooth muscle cells. PMID- 16339492 TI - The fire from within: the biggest Ca2+ channel erupts and dribbles. PMID- 16339493 TI - Endothelial cell IL-8, a new target for adiponectin: implications in vascular protection. PMID- 16339494 TI - Engineering myocardial tissue. AB - To create an artificial heart is one of the most ambitious dreams of the young field of tissue engineering, a dream that, when publicly announced in 1999 (LIFE initiative around M. Sefton), provoked as much compassion as scepticism in the scientific and lay press. Today, it is fair to state that the field is still far away from having built the "bioartificial heart." Nevertheless, substantial progress has been made over the past 10 years, and a realistic perspective exists to create 3-dimensional heart muscle equivalents that may not only serve as experimental models but could also be useful for cardiac regeneration. PMID- 16339498 TI - Conjugated linoleic acid impairs endothelial function. AB - OBJECTIVE: To determine the effect of dietary supplementation with conjugated linoleic acid (CLA) on body mass index (BMI), body fat distribution, endothelial function, and markers of cardiovascular risk. METHODS AND RESULTS: Forty healthy volunteers with BMI >27 kg/m2 were randomized to receive a CLA isomeric mixture or olive oil in a 12-week double-blind study. Subcutaneous body fat and abdominal/hepatic fat content were assessed using skin-fold thicknesses and computed tomography scanning, respectively. Endothelial function was assessed by brachial artery flow-mediated dilatation (FMD). Plasma isoprostanes were measured as an index of oxidative stress. CLA supplementation did not result in a significant change in BMI index or total body fat. There was a significant decrease in limb (-7.8 mm, P<0.001), but not torso skin-fold thicknesses or abdominal or liver fat content. Brachial artery FMD declined (-1.3%, P=0.013), and plasma F2-isoprostanes increased (+91 pg/mL, P=0.042). CONCLUSIONS: A CLA isomeric mixture had at most modest effects on adiposity and worsened endothelial function. On the basis of these results, the use of the isomeric mixture of CLA as an aid to weight loss cannot be recommended. PMID- 16339495 TI - Rho GTPases, statins, and nitric oxide. AB - The lipid-lowering drugs, 3-hydroxy-3-methylgulutaryl-coenzyme A (HMG-CoA) reductase inhibitors or statins, are used in the prevention and treatment of cardiovascular diseases. Recent experimental and clinical studies suggest that statins may exert vascular protective effects beyond cholesterol reduction. For example, statins improve endothelial function by cholesterol-dependent and independent mechanisms. The cholesterol-independent or "pleiotropic" effects of statins include the upregulation and activation of endothelial NO synthase (eNOS). Because statins inhibit an early step in the cholesterol biosynthetic pathway, they also inhibit the synthesis of isoprenoids such as farnesylpyrophosphate and geranylgeranylpyrophosphate, which are important posttranslational lipid attachments for intracellular signaling molecules such as the Rho GTPases. Indeed, decrease in Rho GTPase responses as a consequence of statin treatment increases the production and bioavailability of endothelium derived NO. The mechanism involves, in part, Rho/Rho-kinase (ROCK)-mediated changes in the actin cytoskeleton, which leads to decreases in eNOS mRNA stability. The regulation of eNOS by Rho GTPases, therefore, may be an important mechanism underlying the cardiovascular protective effect of statins. PMID- 16339499 TI - Platelet inhibition by insulin is absent in type 2 diabetes mellitus. AB - OBJECTIVE: ADP-induced P2y12 signaling is crucial for formation and stabilization of an arterial thrombus. We demonstrated recently in platelets from healthy subjects that insulin interferes with Ca2+ increases induced by ADP-P2y1 contact through blockade of the G-protein Gi, and thereby with P2y12-mediated suppression of cAMP. METHODS AND RESULTS: Here we show in patients with type 2 diabetes mellitus (DM2) that platelets have lost responsiveness to insulin leading to increased adhesion, aggregation, and procoagulant activity on contact with collagen. Using Ser473 phosphorylation of protein kinase B as output for insulin signaling, a 2-fold increase is found in insulin-stimulated normal platelets, but in DM platelets there is no significant response. In addition, DM2 platelets show increased P2y12-mediated suppression of cAMP and decreased P2y12 inhibition by the receptor antagonist AR-C69931MX. CONCLUSIONS: The loss of responsiveness to insulin together with increased signaling through P2y12 might explain the hyperactivity of platelets in patients with DM2. PMID- 16339500 TI - Phenotypic modulation of intima and media smooth muscle cells in fatal cases of coronary artery lesion. AB - OBJECTIVE: Characterize the phenotypic features of media and intima coronary artery smooth muscle cells (SMCs) in mildly stenotic plaques, erosions, stable plaques, and in-stent restenosis. METHODS AND RESULTS: Expression of alpha-smooth muscle actin (alpha-SMA), smooth muscle myosin heavy chains (SMMHCs), and smoothelin was investigated by immunohistochemistry followed by morphometric quantification. The cross-sectional area and the expression of cytoskeletal proteins in the media were lower in restenotic lesions and, to a lesser extent, in stable plaques compared with mildly stenotic plaques and erosions. An important expression of alpha-SMA was detected in the intima of the different lesions; moreover, alpha-SMA staining was significantly larger in erosions compared with all other conditions. In the same location, a striking decrease of SMMHCs and a disappearance of smoothelin were observed in all situations. CONCLUSIONS: Medial atrophy is prevalent in restenotic lesions and stable plaques compared with mildly stenotic plaques and erosions. Intimal SMCs of all situations exhibit a phenotypic profile, suggesting that they have modulated into myofibroblasts (MFs). The high accumulation of alpha-SMA-positive MFs in erosions compared with stable plaques correlates with the higher appearance of thrombotic complications in this situation. PMID- 16339501 TI - Smoking, metalloproteinases, and vascular disease. AB - Smoking causes up to 11% of total global cardiovascular deaths. Smoking has numerous effects that may promote atherosclerosis through vascular inflammation and oxidative stress, but the pathogenesis of smoking-related cardiovascular disease remains incompletely understood. The matrix metalloproteinases, a family of endopeptidases that can degrade extracellular matrix components in both physiological and pathophysiological states, play an important role in smoking associated chronic obstructive pulmonary disease, the second leading cause of smoking attributable mortality. Emerging evidence indicates that the matrix metalloproteinases may also contribute to smoking-related vascular disease. Here we discuss the potential relationship between smoking, matrix metalloproteinases, and acceleration of vascular disease. PMID- 16339503 TI - Development of ELISAs measuring the extent of TAFI activation. AB - OBJECTIVE: To date, quantitation of TAFI antigen levels has been mainly focused on "total" antigen levels and has been shown to yield ambiguous results because of the existence of different isoforms and various degrees of activation. Our objective was to develop assays that allow measuring the extent of TAFI activation. METHODS AND RESULTS: A variety of enzyme-linked immunosorbent assays (ELISAs) were evaluated for their preferential reactivity toward TAFI before and after activation, and toward the recombinantly expressed activation peptide. Three ELISAs with distinct reactivities were selected: recognizing either exclusively nonactivated TAFI, the released activation peptide, or exclusively TAFIa (activated TAFI). Evaluation of TAFI activation during clot lysis revealed that decreases of TAFI levels are associated with increases of the released activation peptide and TAFIa levels. In addition, antigenic measurement of TAFIa parallels activity measured by chromogenic assay. Analyzing plasma samples revealed that subjects with hyperlipidemia had significantly higher plasma levels of both the activation peptide (109.2 versus 95.5; P<0.001) and TAFIa (112.1 versus 103.3; P=0.03), and not of TAFI antigen (92.5 versus 87.9; P=0.07) (results in % of plasma pooled from normolipidemic subjects). CONCLUSIONS: ELISAs that allow to measure the extent of TAFI activation were developed. These ELISAs constitute more sensitive markers in studies on the relationship between TAFI and cardiovascular diseases. PMID- 16339502 TI - Role of the estrogen and progestin in hormonal replacement therapy on apolipoprotein A-I kinetics in postmenopausal women. AB - OBJECTIVE: Plasma high-density lipoprotein (HDL) cholesterol levels are inversely correlated with the risk of developing coronary heart disease. Hormonal replacement therapy (HRT) affects plasma HDL cholesterol levels, with estrogen increasing HDL cholesterol levels and progestins blunting this effect. This study was designed to assess the mechanism responsible for these effects. MATERIALS AND METHODS: HDL apolipoprotein A-I (apoA-I) kinetics were studied in 8 healthy postmenopausal women participating in a double-blind, randomized, crossover study comprising 3 phases: placebo, conjugated equine estrogen (CEE) (0.625 mg/d), and CEE plus medroxyprogesterone acetate (MPA) (2.5 mg/d). Compared with placebo, treatment with CEE resulted in an increase in apoA-I pool size (+20%, P<0.01) because of a significant increase in apoA-I production rate (+47%, P<0.05) and no significant changes in apoA-I fractional catabolic rate. Compared with the CEE alone phase, treatment with the CEE plus MPA resulted in an 8% (P<0.02) reduction in apoA-I pool size and a significant reduction in apoA-I production rate (-13%, P<0.04), without changes in apoA-I fractional catabolic rate. CONCLUSIONS: Postmenopausal estrogen replacement increases apoA-I levels and production rate. When progestin is added to estrogen, it opposes these effects by reducing the production of apoA-I. PMID- 16339504 TI - Neurophysiology of perceptual and motor aspects of interception. AB - The interception of moving targets is a complex activity that involves a dynamic interplay of several perceptual and motor processes and therefore involves a rich interaction among several brain areas. Although the behavioral aspects of interception have been studied for the past three decades, it is only during the past decade that neural studies have been focused on this problem. In addition to the interception itself, several neural studies have explored, within that context, the underlying mechanisms concerning perceptual aspects of moving stimuli, such as optic flow and apparent motion. In this review, we discuss the wealth of knowledge that has accumulated on this topic with an emphasis on the results of neural studies in behaving monkeys. PMID- 16339505 TI - Organization and expression of reward in the rodent orbitofrontal cortex. Focus on "orbitofrontal ensemble activity monitors licking and distinguishes among natural rewards". PMID- 16339506 TI - Is spreading depression bad for you? Focus on "repetitive normoxic spreading depression-like events result in cell damage in juvenile hippocampal slice cultures". PMID- 16339507 TI - Immediate-early gene-encoded protein Arc is associated with synaptic delivery of GluR4-containing AMPA receptors during in vitro classical conditioning. AB - The immediate-early gene Arc is rapidly expressed in response to neuronal activity and is thought to be involved in mechanisms of synaptic plasticity. The function of Arc in these processes remains unknown. The present study demonstrates that during an in vitro neural correlate of eyeblink classical conditioning, there is a rapid and transient increase in levels of Arc protein that require activation of N-methyl-d-aspartate receptors. In the early phase of conditioning during conditioned response (CR) acquisition, there is significantly greater colocalization of Arc protein and GluR4-containing AMPA receptors at synaptic sites, however, colocalization of Arc and GluR4 was not observed after later stages of conditioning during CR expression. There was also significantly enhanced coimmunoprecipitation of Arc with GluR4 subunits and actin early in conditioning but not of Arc with NR1 subunits, and these associations declined to control levels in later stages of conditioning. These data suggest a role for Arc protein in the synaptic delivery of GluR4-containing AMPA receptors by interactions with cytoskeletal protein complexes during the acquisition phase of in vitro classical conditioning. PMID- 16339508 TI - Temporal evolution of 2-dimensional direction signals used to guide eye movements. AB - The smooth pursuit system must integrate many local motion measurements into a coherent estimate of target velocity. Several laboratories have studied this integration process using eye movements elicited by targets, such as tilted bars, containing conflicts between local motion signals measured along contours [one dimensional (1D)] and those measured at the bar's endpoints, or terminators [two dimensional (2D)]. The general finding is that 1D signals dominate early responses, whereas later components of the behavior are determined by 2D signals. We studied the dynamics of the integration process in macaque monkeys by systematically varying the relative proportions of 1D and 2D signals and the retinal eccentricities at which they appeared. Predictably, longer bars produced greater and longer-lasting contour-induced deviations. The evolution of the 2D response occurred over a period of 50-400 ms, depending on the relative proportions of 1D and 2D signals. As contours were displaced from the fovea the deviation decreased but much less so for early (1st 40 ms) than for late (subsequent 40 ms) pursuit initiation. These bottom-up effects could be overcome to a limited extent by the top-down influence of predictability. Finally, we observed that when animals were free to track any part of the bar, they spontaneously made short-latency saccades to the terminators on most trials, especially when the bars were tilted. This suggests an increased saliency of moving terminators, particularly when discrepancies exist among local motion signals. PMID- 16339509 TI - How deletions in a model could help explain deletions in the laboratory. PMID- 16339510 TI - Signal regulatory proteins in the immune system. AB - Signal regulatory proteins (SIRPs) constitute a family of transmembrane glycoproteins with extracellular Ig-like domains. Several SIRP family members have thus far been identified on myeloid and other cells in man, mouse, rat, and cattle. In the present study, we provide a description of the SIRP multigene family, including a number of previously undescribed SIRP genes, based on the complete genome sequences of various mammalian and bird species. We discuss this information in the context of the known immunological properties of the individual SIRP family members. Our analysis reveals SIRPs as a diverse multigene family of immune receptors, which includes inhibitory SIRPalpha, activating SIRPbeta, nonsignaling SIRPgamma, and soluble SIRPdelta members. For each species, there appears to be a single inhibitory SIRPalpha member that, upon interaction with the "self" ligand CD47, controls "homeostatic" innate immune effector functions, such as host cell phagocytosis. The activating SIRPbeta proteins show considerable variability in structure and number across species and do not bind CD47. Thus the SIRP family is a rapidly evolving gene family with important roles in immune regulation. PMID- 16339511 TI - A nomenclature for signal regulatory protein family members. PMID- 16339512 TI - Cutting edge: lectin-like transcript 1 is a ligand for the CD161 receptor. AB - Human NK cells and subsets of T cells or NKT cells express the orphan C-type lectin receptor CD161 (NKR-P1A) of unknown function. In contrast to rodents that possess several NKR-P1 genes coding for either activating or inhibitory receptors, the nature of signals delivered by the single human NKR-P1A receptor is still to be clarified. In this article, we show that the lectin-like transcript 1 (LLT1) molecule is a ligand for the CD161 receptor. Engagement of CD161 on NK cells with LLT1 expressed on target cells inhibited NK cell-mediated cytotoxicity and IFN-gamma secretion. Conversely, LLT1/CD161 interaction in the presence of a TCR signal enhanced IFN-gamma production by T cells. These findings identify a novel ligand/receptor pair that differentially regulate NK and T cell functions. PMID- 16339513 TI - Cutting edge: lectin-like transcript-1 is a ligand for the inhibitory human NKR P1A receptor. AB - Increasingly, roles are emerging for C-type lectin receptors in immune regulation. One receptor whose function has remained largely enigmatic is human NKR-P1A (CD161), present on NK cells and subsets of T cells. In this study, we demonstrate that the lectin-like transcript-1 (LLT1) is a physiologic ligand for NKR-P1A. LLT1-containing liposomes bind to NKR-P1A+ cells, and binding is inhibited by anti-NKR-P1A mAb. Additionally, LLT1 activates NFAT-GFP reporter cells expressing a CD3zeta-NKR-P1A chimeric receptor; reciprocally, reporter cells with a CD3zeta-LLT1 chimeric receptor are stimulated by NKR-P1A. Moreover, LLT1 on target cells can inhibit NK cytotoxicity via interactions with NKR-P1A. PMID- 16339514 TI - Cutting edge: FADD is not required for antigen receptor-mediated NF-kappaB activation. AB - Recently, it has been demonstrated that stimulated T cells bearing defects in caspase-8 fail to promote nuclear shuttling of NF-kappaB complexes. Such cells display strikingly similar proliferative and survival defects as T cells lacking Fas-associated death domain protein (FADD) function. We characterized NF-kappaB signaling in T cells bearing a dominant-negative FADD transgene (FADDdd). Whereas FADDdd T cells displayed proliferative defects following activation, these were not a consequence of aberrant NF-kappaB signaling, as measured by IKK/IkappaB phosphorylation and IkappaB degradation. There were no appreciable defects in nuclear translocation of p65/Rel using ImageStream, a flow-based imaging cytometer. Pretreatment with benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, a potent caspase inhibitor, also failed to impede canonical NF-kappaB signaling. Secretion of IL-2 and up-regulation of various activation markers occurred normally. Thus, FADD does not play an essential role in NF-kappaB activation, suggesting an alternative route by which this adaptor promotes the clonal expansion of T cells. PMID- 16339515 TI - Cutting edge: Foxj1 protects against autoimmunity and inhibits thymocyte egress. AB - Previous studies suggest that the forkhead transcription factor Foxj1 inhibits spontaneous autoimmunity in part by antagonizing NF-kappaB activation. To test this hypothesis, we ectopically expressed Foxj1 in the T cells of lupus-prone MRL/lpr mice by backcrossing a CD2-Foxj1 transgene against the MRL/lpr background. Strikingly, CD2-Foxj1-MRL/lpr animals showed a significant reduction in lymphadenopathy, pathogenic autoantibodies, and end-organ disease-but surprisingly, reversion of autoimmunity was not attributable to modulation of NF kappaB. Instead, CD2-Foxj1 transgenic mice exhibited a peripheral T cell lymphopenia, associated with an accumulation of mature single-positive thymocytes. Transgenic thymocytes demonstrated unimpaired lymphoid organ entry in adoptive transfer studies but demonstrated impaired thymic exodus in response to CCL19, apparently independent of CCR7, S1P1, and NF-kappaB. These findings confirm the importance of Foxj1 in the regulation of T cell tolerance but furthermore suggest a novel and specific role for Foxj1 in regulating thymic egress. PMID- 16339516 TI - The scaffolding protein CG-NAP/AKAP450 is a critical integrating component of the LFA-1-induced signaling complex in migratory T cells. AB - T cell migration represents a complex highly coordinated process involving participation of surface receptor/ligand interactions, cytoskeletal rearrangements, and phosphorylation-dependent signaling cascades. Members of the A-kinase anchoring protein (AKAP) family of giant scaffolding proteins can assemble and compartmentalize multiple signaling and structural molecules thereby providing a platform for their targeted positioning and efficient interactions. We characterize here the expression, intracellular distribution, and functional role of the scaffolding protein CG-NAP (centrosome and Golgi localized protein kinase N-associated protein)/AKAP450 in the process of active T cell motility induced via LFA-1 integrins. This protein is predominantly localized at the centrosome and Golgi complex. T cell locomotion triggered by LFA-1 ligation induces redistribution of CG-NAP/AKAP450 along microtubules in trailing cell extensions. Using an original in situ immunoprecipitation approach, we show that CG-NAP/AKAP450 is physically associated with LFA-1 in the multimolecular signaling complex also including tubulin and the protein kinase C beta and delta isoenzymes. CG-NAP/AKAP450 recruitment to this complex was specific for the T cells migrating on LFA-1 ligands, but not on the beta(1) integrin ligand fibronectin. Using the GFP-tagged C-terminal CG-NAP/AKAP450 construct, we demonstrate that expression of the intact CG-NAP/AKAP450 and its recruitment to the LFA-1-associated multimolecular complex is critically important for polarization and migration of T cells induced by this integrin. PMID- 16339517 TI - Silencing human NKG2D, DAP10, and DAP12 reduces cytotoxicity of activated CD8+ T cells and NK cells. AB - Human CD8+ T cells activated and expanded by TCR cross-linking and high-dose IL-2 acquire potent cytolytic ability against tumors and are a promising approach for immunotherapy of malignant diseases. We have recently reported that in vitro killing by these activated cells, which share phenotypic and functional characteristics with NK cells, is mediated principally by NKG2D. NKG2D is a surface receptor that is expressed by all NK cells and transmits an activating signal via the DAP10 adaptor molecule. Using stable RNA interference induced by lentiviral transduction, we show that NKG2D is required for cytolysis of tumor cells, including autologous tumor cells from patients with ovarian cancer. We also demonstrated that NKG2D is required for in vivo antitumor activity. Furthermore, both activated and expanded CD8+ T cells and NK cells use DAP10. In addition, direct killing was partially dependent on the DAP12 signaling pathway. This requirement by activated and expanded CD8+ T cells for DAP12, and hence stimulus from a putative DAP12-partnered activating surface receptor, persisted when assayed by anti-NKG2D Ab-mediated redirected cytolysis. These studies demonstrated the importance of NKG2D, DAP10, and DAP12 in human effector cell function. PMID- 16339519 TI - Identification of T cell-restricted genes, and signatures for different T cell responses, using a comprehensive collection of microarray datasets. AB - We used a comprehensive collection of Affymetrix microarray datasets to ascertain which genes or molecules distinguish the known major subsets of human T cells. Our strategy allowed us to identify the genes expressed in most T cell subsets: TCR alphabeta+ and gammadelta+, three effector subsets (Th1, Th2, and T follicular helper cells), T central memory, T effector memory, activated T cells, and others. Our genechip dataset also allowed for identification of genes preferentially or exclusively expressed by T cells, compared with numerous non-T cell leukocyte subsets profiled. Cross-comparisons between microarray datasets revealed important features of certain subsets. For instance, blood gammadelta T cells expressed no unique gene transcripts, but did differ from alphabeta T cells in numerous genes that were down-regulated. Hierarchical clustering of all the genes differentially expressed between T cell subsets enabled the identification of precise signatures. Moreover, the different T cell subsets could be distinguished at the level of gene expression by a smaller subset of predictor genes, most of which have not previously been associated directly with any of the individual subsets. T cell activation had the greatest influence on gene regulation, whereas central and effector memory T cells displayed surprisingly similar gene expression profiles. Knowledge of the patterns of gene expression that underlie fundamental T cell activities, such as activation, various effector functions, and immunological memory, provide the basis for a better understanding of T cells and their role in immune defense. PMID- 16339518 TI - CD80 cytoplasmic domain controls localization of CD28, CTLA-4, and protein kinase Ctheta in the immunological synapse. AB - The binding of costimulatory ligand CD80 to CD28 or CTLA-4 on T cells plays an important role in the regulation of the T cell response. We have examined the role of the cytoplasmic domain of CD80 in murine T cell costimulation and its organization in the immunological synapse (IS). Removal of CD80 cytoplasmic tail decreased its effectiveness in costimulating T cell proliferative response and early IL-2 production in response to agonist MHC-peptide complexes. Immunofluorescent study showed a decreased tailless CD80 accumulation in the IS of naive T cells. The two forms of CD80 accumulated differently at the IS; the tailless CD80 was colocalized with the TCR whereas the full-length CD80 was segregated from the TCR. In addition, we showed that CD80, CD28, and protein kinase Ctheta colocalized in the presence or absence of the CD80 cytoplasmic tail. Thus, the cytoplasmic tail of CD80 regulates its spatial localization at the IS and that of its receptors and T cell signaling molecules such as protein kinase Ctheta, and thereby facilitates full T cell activation. PMID- 16339520 TI - Ligation of CD28 by its natural ligand CD86 in the absence of TCR stimulation induces lipid raft polarization in human CD4 T cells. AB - Stimulation of resting CD4 T cells with anti-CD3/CD28-coated beads leads to rapid polarization of lipid rafts (LRs). It has been postulated that a major role of costimulation is to facilitate LR aggregation. CD86 is up-regulated or expressed aberrantly on immune cells in a wide array of autoimmune and infectious diseases. Using an Ig fusion with the extracellular domain of CD86 (CD86Ig) bound to a magnetic bead or K562 cells expressing CD86, we demonstrated that ligation of CD28 by its natural ligand, but not by Ab, induced polarization of LRs at the cell-bead interface of fresh human CD4 T cells in the absence of TCR ligation. This correlated with activation of Vav-1, increase of the intracellular calcium concentration, and nuclear translocation of NF-kappaB p65, but did not result in T cell proliferation or cytokine production. These studies show, for the first time, that LR polarization can occur in the absence of TCR triggering, driven solely by the CD28/CD86 interaction. This result has implications for mechanisms of T cell activation. Abnormalities in this process may alter T and B cell tolerance and susceptibility to infection. PMID- 16339521 TI - Targeting LFA-1 and cd154 suppresses the in vivo activation and development of cytolytic (cd4-Independent) CD8+ T cells. AB - Short-term immunotherapy targeting both LFA-1 and CD40/CD154 costimulation produces synergistic effects such that long-term allograft survival is achieved in the majority of recipients. This immunotherapeutic strategy has been reported to induce the development of CD4+ regulatory T cells. In the current study, the mechanisms by which this immunotherapeutic strategy prevents CD8+ T cell dependent hepatocyte rejection in CD4 knockout mice were examined. Combined blockade of LFA-1 and CD40/CD154 costimulation did not influence the overall number or composition of inflammatory cells infiltrating the liver where transplanted hepatocytes engraft. Expression of T cell activation markers CD43, CD69, and adhesion molecule CD103 by liver-infiltrating cells was suppressed in treated mice with long-term hepatocellular allograft survival compared to liver infiltrating cells of untreated rejector mice. Short-term immunotherapy with anti LFA-1 and anti-CD154 mAb also abrogated the in vivo development of alloreactive CD8+ cytotoxic T cell effectors. Treated mice with long-term hepatocyte allograft survival did not reject hepatocellular allografts despite adoptive transfer of naive CD8+ T cells. Unexpectedly, treated mice with long-term hepatocellular allograft survival demonstrated prominent donor-reactive delayed-type hypersensitivity responses, which were increased in comparison to untreated hepatocyte rejectors. Collectively, these findings support the conclusion that short-term immunotherapy with anti-LFA-1 and anti-CD154 mAbs induces long-term survival of hepatocellular allografts by interfering with CD8+ T cell activation and development of CTL effector function. In addition, these recipients with long term hepatocellular allograft acceptance show evidence of immunoregulation which is not due to immune deletion or ignorance and is associated with early development of a novel CD8+CD25high cell population in the liver. PMID- 16339522 TI - IL-21 induces differentiation of human naive and memory B cells into antibody secreting plasma cells. AB - IL-21 is a type I cytokine that influences the function of T cells, NK cells, and B cells. In this study, we report that IL-21 plays a major role in stimulating the differentiation of human B cells. When human B cells were stimulated through the BCR, IL-21 induced minimal proliferation, IgD down-modulation, and small numbers of plasma cells. In contrast, after CD40 engagement, IL-21 induced extensive proliferation, class switch recombination (CSR), and plasma cell differentiation. Upon cross-linking both BCR and CD40, IL-21 induced the largest numbers of plasma cells. IL-21 drove both postswitch memory cells as well as poorly responsive naive cord blood B cells to differentiate into plasma cells. The effect of IL-21 was more potent than the combination of IL-2 and IL-10, especially when responsiveness of cord blood B cells was examined. IL-21 costimulation potently induced the expression of both B lymphocyte-induced maturation protein-1 (BLIMP-1) and activation-induced cytidine deaminase as well as the production of large amounts of IgG from B cells. Despite the induction of activation-induced cytidine deaminase and CSR, IL-21 did not induce somatic hypermutation. Finally, IL-2 enhanced the effects of IL-21, whereas IL-4 inhibited IL-21-induced plasma cell differentiation. Taken together, our data show that IL-21 plays a central role in CSR and plasma cell differentiation during T cell-dependent B cell responses. PMID- 16339523 TI - Increased expression of genes linked to FcepsilonRI Signaling and to cytokine and chemokine production in Lyn-deficient mast cells. AB - Cross-linking the high-affinity IgE receptor, FcepsilonRI, on mast cells activates signaling pathways leading to the release of preformed inflammatory mediators and the production of cytokines and chemokines associated with allergic disorders. Bone marrow-derived mast cells (BMMCs) from Lyn-deficient (Lyn-/-) mice are hyperresponsive to FcepsilonRI cross-linking with multivalent Ag. Previous studies linked the hyperresponsive phenotype in part to increased Fyn kinase activity and reduced SHIP phosphatase activity in the Lyn-/- BMMCs in comparison with wild-type (WT) cells. In this study, we compared gene expression profiles between resting and Ag-activated WT and Lyn-/- BMMCs to identify other factors that may contribute to the hyperresponsiveness of the Lyn-/- cells. Among genes implicated in the positive regulation of FcepsilonRI signaling, mRNA for the tyrosine kinase, Fyn, and for several proteins contributing to calcium regulation are more up-regulated following Ag stimulation in Lyn-/- BMMCs than in WT BMMCs. Conversely, mRNA for the low-affinity IgG receptor (FcgammaRIIB), implicated in negative regulation of FcepsilonRI-mediated signaling, is more down regulated in Ag-stimulated Lyn-/- BMMCs than in WT BMMCs. Genes coding for proinflammatory cytokines and chemokines (IL-4, IL-6, IL-13, CSF, CCL1, CCL3, CCL5, CCL7, CCL9, and MIP1beta) are all more highly expressed in Ag-stimulated Lyn-/- mast cells than in WT cells. These microarray data identify Lyn as a negative regulator in Ag-stimulated BMMCs of the expression of genes linked to FcepsilonRI signaling and also to the response pathways that lead to allergy and asthma. PMID- 16339524 TI - CD18 is required for optimal development and function of CD4+CD25+ T regulatory cells. AB - CD4+CD25+ T regulatory (Treg) cells inhibit immunopathology and autoimmune disease in vivo. CD4+CD25+ Treg cells' capacity to inhibit conventional T cells in vitro is dependent upon cell-cell contact; however, the cell surface molecules mediating this cell:cell contact have not yet been identified. LFA-1 (CD11a/CD18) is an adhesion molecule that plays an established role in T cell-mediated cell contact and in T cell activation. Although expressed at high levels on murine CD4+CD25+ Treg cells, the role of LFA-1 in these cells has not been defined previously. We hypothesized that LFA-1 may play a role in murine CD4+CD25+ Treg function. To evaluate this, we analyzed LFA-1-deficient (CD18-/-) CD4+CD25+ T cells. We show that CD18-/- mice demonstrate a propensity to autoimmunity. Absence of CD18 led to diminished CD4+CD25+ T cell numbers and affected both thymic and peripheral development of these cells. LFA-1-deficient CD4+CD25+ T cells were deficient in mediating suppression in vitro and in mediating protection from colitis induced by the transfer of CD4+CD25- T cells into lymphopenic hosts. Therefore, we define a crucial role for CD18 in optimal CD4+CD25+ Treg development and function. PMID- 16339525 TI - Cooperation of invariant NKT cells and CD4+CD25+ T regulatory cells in the prevention of autoimmune myasthenia. AB - CD1d-restricted NKT cells and CD4+CD25+ regulatory T (Treg) cells are thymus derived subsets of regulatory T cells that have an important role in the maintenance of self-tolerance. Whether NKT cells and Treg cells cooperate functionally in the regulation of autoimmunity is not known. We have explored this possibility in experimental autoimmune myasthenia gravis (EAMG), an animal model of human myasthenia gravis, induced by immunization of C57BL/6 mice with the autoantigen acetylcholine receptor. We have demonstrated that activation of NKT cells by a synthetic glycolipid agonist of NKT cells, alpha galactosylceramide (alpha-GalCer), inhibits the development of EAMG. alpha-GalCer administration in EAMG mice increased the size of the Treg cell compartment, and augmented the expression of foxp3 and the potency of CD4+CD25+ cells to inhibit proliferation of autoreactive T cells. Furthermore, alpha-GalCer promoted NKT cells to transcribe the IL-2 gene and produce IL-2 protein. Depletion of CD25+ cells or neutralization of IL-2 reduced the therapeutic effect of alpha-GalCer in this model. Thus, alpha-GalCer-activated NKT cells can induce expansion of CD4+CD25+ Treg cells, which in turn mediate the therapeutic effects of alpha GalCer in EAMG. Induced cooperation of NKT cells and Treg cells may serve as a superior strategy to treat autoimmune disease. PMID- 16339526 TI - CD4+ T cell responses elicited by different subsets of human skin migratory dendritic cells. AB - Skin dendritic cells (DC) are professional APC critical for initiation and control of adaptive immunity. In the present work we have analyzed the CD4+ T cell stimulatory function of different subsets of DC that migrate spontaneously from human skin explants, including CD1a+CD14- Langerhans' cells (LC), CD1a-CD14- dermal DC (DDC), and CD1a-CD14+ LC precursors. Skin migratory DC consisted of APC at different stages of maturation-activation that produced IL-10, TGF-beta1, IL 23p19, and IL-12p40, but did not release IL-12p70 even after exposure to DC1 driving stimuli. LC and DDC migrated as mature/activated APC able to stimulate allogeneic naive CD4+ T cells and to induce memory Th1 cells in the absence of IL 12p70. The potent CD4+ T cell stimulatory function of LC and DDC correlated with their high levels of expression of MHC class II, adhesion, and costimulatory molecules. The Th1-biasing function of LC and DDC depended on their ability to produce IL-23. By contrast, CD1a-CD14+ LC precursors migrated as immature semimature APC and were weak stimulators of allogeneic naive CD4+ T cells. However, and opposite of a potential tolerogenic role of immature DC, the T cell allostimulatory and Th1-biasing function of CD14+ LC precursors increased significantly by augmenting their cell number, prolonging the time of interaction with responding T cells, or addition of recombinant human IL-23 in MLC. The data presented in this study provide insight into the function of the complex network of skin-resident DC that migrate out of the epidermis and dermis after cutaneous immunizations, pathogen infections, or allograft transplantation. PMID- 16339527 TI - Retinoid x receptor agonists increase bcl2a1 expression and decrease apoptosis of naive T lymphocytes. AB - Vitamin A affects many aspects of T lymphocyte development and function. The vitamin A metabolites all-trans- and 9-cis-retinoic acid regulate gene expression by binding to the retinoic acid receptor (RAR), while 9-cis-retinoic acid also binds to the retinoid X receptor (RXR). Naive DO11.10 T lymphocytes expressed mRNA and protein for RAR-alpha, RXR-alpha, and RXR-beta. DNA microarray analysis was used to identify RXR-responsive genes in naive DO11.10 T lymphocytes treated with the RXR agonist AGN194204. A total of 128 genes was differentially expressed, including 16 (15%) involved in cell growth or apoptosis. Among these was Bcl2a1, an antiapoptotic Bcl2 family member. Quantitative real-time PCR analysis confirmed this finding and demonstrated that Bcl2a1 mRNA expression was significantly greater in nonapoptotic than in apoptotic T lymphocytes. The RXR agonist 9-cis-retinoic acid also increased Bcl2a1 expression, although all-trans retinoic acid and ligands for other RXR partner receptors did not. Treatment with AGN194204 and 9-cis-retinoic acid significantly decreased apoptosis measured by annexin V staining but did not affect expression of Bcl2 and Bcl-xL. Bcl2a1 promoter activity was examined using a luciferase promoter construct. Both AGN194204 and 9-cis-retinoic acid significantly increased luciferase activity. In summary, these data demonstrate that RXR agonists increase Bcl2a1 promoter activity and increase expression of Bcl2a1 in naive T lymphocytes but do not affect Bcl2 and Bcl-xL expression in naive T lymphocytes. Thus, this effect on Bcl2a1 expression may account for the decreased apoptosis seen in naive T lymphocytes treated with RXR agonists. PMID- 16339528 TI - Identification of specific proteins and peptides in Mycobacterium leprae suitable for the selective diagnosis of leprosy. AB - Diagnosis of leprosy is a major obstacle to disease control and has been compromised in the past due to the lack of specific reagents. We have used comparative genome analysis to identify genes that are specific to Mycobacterium leprae and tested both recombinant proteins and synthetic peptides from a subset of these for immunological reactivity. Four unique recombinant proteins (ML0008, ML0126, ML1057, and ML2567) and a panel of 58 peptides (15 and 9 mer) were tested for IFN-gamma responses in PBMC from leprosy patients and contacts, tuberculosis patients, and endemic and nonendemic controls. The responses to the four recombinant proteins gave higher levels of IFN-gamma production, but less specificity, than the peptides. Thirty-five peptides showed IFN-gamma responses only in the paucibacillary leprosy and household contact groups, with no responses in the tuberculosis or endemic control groups. High frequencies of IFN gamma-producing CD4+ and CD8+ T cells specific for the 15- and 9-mer peptides were observed in the blood of a paucibacillary leprosy patient. 9-mer peptides preferentially activated CD8+ T cells, while the 15-mer peptides were efficient in inducing responses in both the CD4+ and CD8+ T cell subsets. Four of the six 9 mer peptides tested showed promising specificity, indicating that CD8+ T cell epitopes may also have diagnostic potential. Those peptides that provide specific responses in leprosy patients from an endemic setting could potentially be developed into a rapid diagnostic test for the early detection of M. leprae infection and epidemiological surveys of the incidence of leprosy, of which little is known. PMID- 16339529 TI - Long-term MHC class II presentation of the EBV lytic protein BHRF1 by EBV latently infected b cells following capture of BHRF1 antigen. AB - Although T lymphocytes are considered essential for the control of EBV infection, it remains uncertain how this control occurs. We previously reported unexpected killing of EBV-transformed B-lymphoblastoid cells (LCLs) that did not express BHRF1 by CD4+ T cells specific for BHRF1, an EBV lytic cycle protein. Using LCLs transformed with an EBV mutant, in which the BHRF1 gene was deleted, we showed that killing of latently infected cells through the recognition of a protein produced during the lytic cycle is due to transfer of BHRF1 from lytically infected to latently infected cells, which occurs in culture. Accordingly, LCLs efficiently presented exogenous BHRF1 protein. Furthermore, we present evidence for persistence of captured BHRF1 Ag for several days. Due to this long-term persistence, repeated loading of suboptimal amounts of BHRF1 led to accumulation of BHRF1 Ags in LCLs and, ultimately, to their optimal recognition by BHRF1 specific CD4+ T cells. These results unveil an MHC class II-dependent pathway that could be important for the control of EBV latent infection through recognition of lytic cycle Ags. PMID- 16339530 TI - Protein tyrosine phosphatase alpha regulates Fyn activity and Cbp/PAG phosphorylation in thymocyte lipid rafts. AB - A role for the receptor protein tyrosine phosphatase alpha (PTPalpha) in immune cell function and regulation of Src family kinases was investigated using thymocytes from PTPalpha-deficient mice. PTPalpha-null thymocytes develop normally, but unstimulated PTPalpha-/- cells exhibit increased tyrosine phosphorylation of specific proteins, increased Fyn activity, and hyperphosphorylation of Cbp/PAG that promotes its association with C-terminal Src kinase. Elevated Fyn activity in the absence of PTPalpha is due to enhanced phosphorylation of Fyn tyrosines 528 and 417. Some PTPalpha is localized in lipid rafts of thymocytes, and raft-associated Fyn is specifically activated in PTPalpha-/- cells. PTPalpha is not a Cbp/PAG phosphatase, because it is not required for Cbp/PAG dephosphorylation in unstimulated or anti-CD3-stimulated thymocytes. Together, our results indicate that PTPalpha, likely located in lipid rafts, regulates the activity of raft Fyn. In the absence of PTPalpha this population of Fyn is activated and phosphorylates Cbp/PAG to enhance association with C-terminal Src kinase. Although TCR-mediated tyrosine phosphorylation was apparently unaffected by the absence of PTPalpha, the long-term proliferative response of PTPalpha-/- thymocytes was reduced. These findings indicate that PTPalpha is a component of the complex Src family tyrosine kinase regulatory network in thymocytes and is required to suppress Fyn activity in unstimulated cells in a manner that is not compensated for by the major T cell PTP and SFK regulator, CD45. PMID- 16339531 TI - Quantifying the contribution of defective ribosomal products to antigen production: a model-based computational analysis. AB - Antigenic peptides (epitopes) presented on the cell surface by MHC class I molecules derive from proteolytic degradation of endogenous proteins. Some recent studies have proposed that the majority of epitopes stem from so-called defective ribosomal products (DRiPs), i.e., freshly synthesized proteins that are unable to adopt the native conformation and thus undergo immediate degradation. However, a reliable computational analysis of the data underlying this hypothesis was lacking so far. Therefore, we have applied kinetic modeling to derive from existing kinetic data (Princiotta et al. 2003, Immunity 18, 343-354) the rates of the major processes involved in the cellular protein turnover and MHC class I mediated Ag presentation. From our modeling approach, we conclude that in these experiments 1) the relative share of DRiPs in the total protein synthesis amounted to approximately 10% thus being much lower than reported so far, 2) DRiPs may become the decisive source of epitopes within an early phase after onset of the synthesis of a long-lived (e.g., virus derived) protein, and 3) inhibition of protein synthesis by the translation inhibitor cycloheximide appears to be paralleled with an instantaneous decrease of protein degradation down to approximately 1/3 of the normal value. PMID- 16339532 TI - The role of low molecular weight thiols in T lymphocyte proliferation and IL-2 secretion. AB - Glutathione (GSH) is an abundant intracellular tripeptide that has been implicated as an important regulator of T cell proliferation. The effect of pharmacological regulators of GSH and other thiols on murine T cell signaling, proliferation, and intracellular thiol levels was examined. l-Buthionine-S,R sulfoximine (BSO), an inhibitor of GSH synthesis, markedly reduced GSH levels and blocked T cell proliferation without significant effect on cell viability. N acetylcysteine markedly enhanced T cell proliferation without affecting GSH levels. Cotreatment of T cells with N-acetylcysteine and BSO failed to restore GSH levels, but completely restored the proliferative response. Both 2-ME and l cysteine also reversed the BSO inhibition of T cell proliferation. Intracellular l-cysteine levels were reduced with BSO treatment and restored with cotreatment with NAC or l-cysteine. However, 2-ME completely reversed the BSO inhibition of proliferation without increasing intracellular cysteine levels. Therefore, neither GSH nor cysteine is singularly critical in limiting T cell proliferation. Reducing equivalents from free thiols were required because oxidation of the thiol moiety completely abolished the effect. Furthermore, BSO did not change the expression of surface activation markers, but effectively blocked IL-2 and IL-6 secretion. Importantly, exogenous IL-2 completely overcame BSO-induced block of T cell proliferation. These results demonstrate that T cell proliferation is regulated by thiol-sensitive pathway involving IL-2. PMID- 16339533 TI - The sphingosine 1-phosphate receptor agonist FTY720 differentially affects the sequestration of CD4+/CD25+ T-regulatory cells and enhances their functional activity. AB - The sphingosine 1-phosphate (S1P) receptor agonist FTY720 is well known for its immunomodulatory activity, sequestering lymphocytes from blood and spleen into secondary lymphoid organs and thereby preventing their migration to sites of inflammation. Because inflammation is critically dependent on a balance between Ag-specific Th/effector cells and T-regulatory cells, we investigated the effect of FTY720 on T-regulatory cell trafficking and functional activity. An increased number of CD4+/CD25+ T cells was found in blood and spleens of FTY720-treated mice, and transfer of these cells resulted in a significantly more pronounced accumulation in spleens but not lymph nodes after treatment, suggesting that this compound differentially affects the homing properties of T-regulatory cells compared with other T cell subsets. Indeed, CD4+/CD25+ T cells express lower levels of S1P1 and S1P4 receptors and demonstrate a reduced chemotactic response to S1P. Moreover, analysis of the functional response of FTY720-treated CD4+/CD25+ T cells revealed an increased suppressive activity in an in vitro Ag specific proliferation assay. This correlated with enhanced function in vivo, with T-regulatory cells obtained from FTY720-treated mice being able to suppress OVA-induced airway inflammation. Thus, FTY720 differentially affects the sequestration of T-regulatory cells and importantly, increases the functional activity of T-regulatory cells, suggesting that it may have disease-modifying potential in inflammatory disorders. PMID- 16339534 TI - Stabilized beta-catenin extends thymocyte survival by up-regulating Bcl-xL. AB - CD4+CD8+ double-positive (DP) thymocytes, which are extremely sensitive to apoptosis, specifically up-regulate Bcl-xL to extend their lifespan. Deletion of the Bcl-xL gene leads to premature apoptosis of the thymocytes. In this study, we show that stabilization of beta-catenin, a critical coactivator for T cell factor (TCF), enhances DP thymocyte survival via up-regulating Bcl-xL. Spontaneous or glucocorticoid-induced thymocyte apoptosis was associated with reduced levels of beta-catenin and Bcl-xL. Transgenic expression of a stabilized beta-catenin protected DP thymocytes from both spontaneous and glucocorticoid-induced apoptosis, resulting in significantly increased thymic cellularity. Compared with the wild-type mice, both protein and transcript levels of Bcl-xL were significantly increased in thymocytes of beta-catenin transgenic mice. In addition, TCF-1 as well as beta-catenin were able to stimulate transcriptional activity of the reporter driven by a Bcl-xL promoter. beta-Catenin/TCF is thus able to act as a signal to up-regulate Bcl-xL levels in DP thymocytes, resulting in their enhanced survival. PMID- 16339535 TI - The inhibitory receptor IRp60 (CD300a) is expressed and functional on human mast cells. AB - Mast cell-mediated responses are likely to be regulated by the cross talk between activatory and inhibitory signals. We have screened human cord blood mast cells for recently characterized inhibitory receptors expressed on NK cells. We found that IRp60, an Ig superfamily member, is expressed on human mast cells. On NK cells, IRp60 cross-linking leads to the inhibition of cytotoxic activity vs target cells in vitro. IRp60 is constitutively expressed on mast cells but is down-regulated in vitro by the eosinophil proteins major basic protein and eosinophil-derived neurotoxin. An immune complex-mediated cross-linking of IRp60 led to inhibition of IgE-induced degranulation and stem cell factor-mediated survival via a mechanism involving tyrosine phosphorylation, phosphatase recruitment, and termination of cellular calcium influx. To evaluate the role of IRp60 in regulation of allergic responses in vivo, a murine model of allergic peritonitis was used in which the murine homolog of IRp60, LMIR1, was neutralized in BALB/c mice by mAbs. This neutralization led to a significantly augmented release of inflammatory mediators and eosinophilic infiltration. These data demonstrate a novel pathway for the regulation of human mast cell function and allergic responses, indicating IRp60 as a candidate target for future treatment of allergic and mast cell-associated diseases. PMID- 16339536 TI - The cytotoxicity receptor CRACC (CS-1) recruits EAT-2 and activates the PI3K and phospholipase Cgamma signaling pathways in human NK cells. AB - The CD2-like receptor-activating cytotoxic cell (CRACC) is a cell surface receptor of the CD2 family that triggers NK cell-mediated cytotoxicity through an undefined signaling pathway. CRACC contains cytoplasmic tyrosine-based motifs, immunoreceptor tyrosine-based switch motifs, which resemble those found in the NK cell receptor 2B4. In 2B4, these motifs recruit the adaptor signaling lymphocytic activation molecule-associated protein (SAP), which initiates a signaling cascade mediating cytotoxicity. However, CRACC does not recruit SAP. In this study, we demonstrate that, upon activation, CRACC associates with a homolog of SAP, Ewing's sarcoma's/FLI1-activated transcript 2 (EAT-2), in human NK cells. We show that association of EAT-2 induces the phosphorylation of CRACC and that this process is partially reduced by a pharmacological inhibitor of Src kinases. We identify PLCgamma1, PLCgamma2, and PI3K as the major signaling mediators downstream of CRACC/EAT-2 implicated in NK cell-mediated cytotoxicity. Moreover, EAT-2 also associates with 2B4 predominantly in resting NK cells, whereas SAP preferentially binds 2B4 upon activation. These results outline a new signaling pathway that triggers CRACC-mediated cytotoxicity and modulates 2B4-mediated activation. PMID- 16339537 TI - IL-2 regulates perforin and granzyme gene expression in CD8+ T cells independently of its effects on survival and proliferation. AB - Perforin and the serine protease granzymes are key effectors of CD8+ T cell granule-mediated cytotoxicity, but the requirements for their expression remain largely undefined. We show in this study that IL-2 increased the expression of perforin and granzyme A, B, and C mRNA; intracellular granzyme B protein levels; and cytolytic function in a dose-dependent manner during primary activation of murine CD8+ T cells in vitro. Two approaches showed that these responses were not a consequence of the effects of IL-2 on cell survival and proliferation. First, IL-2 enhancement of perforin and granzyme expression was equivalent in CD8+ T cells from wild-type and bcl-2 transgenic mice, although only the latter cells survived in low concentrations or the absence of added IL-2. This property of bcl 2 transgenic T cells also allowed the demonstration that induction of granzyme A, B, and C mRNA and granzyme B protein required exogenous IL-2, whereas induction of perforin and IFN-gamma expression did not. Second, analysis of perforin and granzyme mRNA levels in cells separated according to division number using the dye CFSE showed that the effects of IL-2 were unrelated to division number. Together, these findings indicate that IL-2 can directly regulate perforin and granzyme gene expression in CD8+ T cells independently of its effects on cell survival and proliferation. PMID- 16339538 TI - Complement component C3d-antigen complexes can either augment or inhibit B lymphocyte activation and humoral immunity in mice depending on the degree of CD21/CD19 complex engagement. AB - C3d can function as a molecular adjuvant by binding CD21 and thereby enhancing B cell activation and humoral immune responses. However, recent studies suggest both positive and negative roles for C3d and the CD19/CD21 signaling complex in regulating humoral immunity. To address whether signaling through the CD19/CD21 complex can negatively regulate B cell function when engaged by physiological ligands, diphtheria toxin (DT)-C3d fusion protein and C3dg-streptavidin (SA) complexes were used to assess the role of CD21 during BCR-induced activation and in vivo immune responses. Immunization of mice with DT-C3d3 significantly reduced DT-specific Ab responses independently of CD21 expression or signaling. By contrast, SA-C3dg tetramers dramatically enhanced anti-SA responses when used at low doses, whereas 10-fold higher doses did not augment immune responses, except in CD21/35-deficient mice. Likewise, SA-C3dg (1 microg/ml) dramatically enhanced BCR-induced intracellular calcium concentration ([Ca2+]i) responses in vitro, but had no effect or inhibited [Ca2+]i responses when used at 10- to 50-fold higher concentrations. SA-C3dg enhancement of BCR-induced [Ca2+]i responses required CD21 and CD19 expression and resulted in significantly enhanced CD19 and Lyn phosphorylation, with enhanced Lyn/CD19 associations. BCR-induced CD22 phosphorylation and Src homology 2 domain-containing protein tyrosine phosphatase 1/CD22 associations were also reduced, suggesting abrogation of negative regulatory signaling. By contrast, CD19/CD21 ligation using higher concentrations of SA-C3dg significantly inhibited BCR-induced [Ca2+]i responses and inhibited CD19, Lyn, CD22, and Syk phosphorylation. Therefore, C3d may enhance or inhibit Ag-specific humoral immune responses through both CD21-dependent and -independent mechanisms depending on the concentration and nature of the Ag-C3d complexes. PMID- 16339539 TI - Fas ligand induces cell-autonomous IL-23 production in dendritic cells, a mechanism for Fas ligand-induced IL-17 production. AB - Fas ligand (FasL) has the potential to induce inflammation accompanied by massive neutrophil infiltration. We previously reported that FasL rapidly induces the production of various inflammatory cytokines including IL-1beta and IL-17. In this study, we investigated the mechanism of the FasL-induced IL-17 production. We found that the culture supernatant of mouse resident peritoneal exudate cells (PEC) cocultured with FasL-expressing tumor (FFL) cells induced IL-17 production in freshly isolated resident PEC. Anti-IL-1beta Ab strongly inhibited the IL-17 inducing activity. However, rIL-1beta by itself induced only weak IL-17 production. Intriguingly, anti-IL-12 Ab but not an IL-15-neutralizing agent, IL15R-Fc, strongly inhibited the FasL-induced IL-17-inducing activity. IL-23, which shares the p40 subunit with IL-12, but not IL-12 itself, induced IL-17 production synergistically with IL-1beta in resident PEC. FasL induced the production of IL-23 in PEC in vivo and in vitro, and IL-17 production following the i.p. injection of FFL cells was severely impaired in p40-/- mice, indicating that IL-23 plays an important role in the FasL-induced IL-17 production. FFL also induced the production of IL-23 in bone marrow- or PEC-derived dendritic cells (DCs). Finally, FasL induced only weak p40 production in a mixture of p40-/- and Fas-/- DC, indicating that FasL induces IL-23 production in DC mainly in a cell autonomous manner. PMID- 16339540 TI - Disparate regulation and function of the class A scavenger receptors SR-AI/II and MARCO. AB - The macrophage class A scavenger receptors, macrophage receptor with a collagenous structure (MARCO) and type I/II class A scavenger receptor (SR AI/II), share structural features and roles in host defense, but little is known about their regulation and signaling properties. Ligation of MARCO on mouse thioglycollate-elicited peritoneal macrophages (PEMs) with immobilized mAb costimulated IL-12 production, in contrast to previously reported inhibition by SR-AI/II. PEMs from MARCO-deficient mice exhibited 2.7 times lower IL-12 production in responses to stimulation with LPS and IFN-gamma and lack of significant IL-12 production on stimulation with LPS alone. Conversely, SR-AI/II deficient PEMs produced 2.4 and 1.8 times more IL-12 than wild-type PEMs in response to LPS or LPS and IFN-gamma, respectively. Corresponding differences in regulation of SR-A and MARCO expression were also observed. Th1 adjuvants (LPS, a CpG motif-containing oligodeoxynucleotide (CpG-ODN), IL-12, and GM-CSF) increased, whereas Th2-polarizing factors (IL-4, M-CSF, and non-CpG ODN) decreased expression of MARCO on J774 macrophage-like cells. Expression of SR-A was regulated in the opposite manner to MARCO or not affected. Whereas MARCO was involved in opsonin-independent phagocytosis in CpG-ODN-pretreated but not in IL 4-pretreated J774 cells, anti-SR-A Abs inhibited particle uptake in untreated and IL-4-pretreated but not in CpG-ODN-pretreated cells. SR-A and MARCO are regulated differently and mediate distinct negative and positive effects on IL-12 production in macrophages. These differences may contribute to sustained Th1 or Th2 polarization of ongoing immune responses. PMID- 16339541 TI - CD43 functions as a ligand for E-Selectin on activated T cells. AB - E-selectin, an inducible cell adhesion molecule expressed on endothelial cells, mediates the rolling on endothelium of leukocytes expressing E-selectin ligands, such as neutrophils and activated T cells. Although previous studies using mice lacking P-selectin glycoprotein ligand-1 (PSGL-1) have indicated that PSGL-1 on Th1 cells functions as an E-selectin ligand, the molecular nature of E-selectin ligands other than PSGL-1 remains unknown. In this study, we show that a 130-kDa glycoprotein was precipitated by an E-selectin-IgG chimera from mouse Th1 cells. This protein was cleaved by O-sialoglycoprotein endopeptidase and required sialic acid for E-selectin binding. The mAb 1B11, which recognizes the 130-kDa glycoform of CD43, recognized the 130-kDa band in the E-selectin-IgG precipitate. In addition, immunoprecipitation of the E-selectin-IgG precipitate with 1B11 depleted the 130-kDa protein, further confirming its identity as CD43. CD43 was also precipitated with E-selectin-IgG from cultured human T cells. E-selectin dependent cell rolling on CD43 was observed under flow conditions using a CD43 IgG chimera generated in Chinese hamster ovary cells expressing alpha-1,3 fucosyltransferase VII and a core 2 beta-1,6-N-acetylglucosaminyltransferase. These results suggest that CD43, when modified by a specific set of glycosyltranferases, can function as an E-selectin ligand and therefore potentially mediate activated T cell migration into inflamed sites. PMID- 16339542 TI - Human CD4+ T cells express TLR5 and its ligand flagellin enhances the suppressive capacity and expression of FOXP3 in CD4+CD25+ T regulatory cells. AB - Germline encoded pattern recognition receptors, such as TLRs, provide a critical link between the innate and adaptive immune systems. There is also evidence to suggest that pathogen-associated molecular patterns may have the capacity to modulate immune responses via direct effects on CD4+ T cells. Given the key role of both CD4+CD25+ T regulatory (Treg) cells and the TLR5 ligand flagellin in regulating mucosal immune responses, we investigated whether TLR5 may directly influence T cell function. We found that both human CD4+CD25+ Treg and CD4+CD25- T cells express TLR5 at levels comparable to those on monocytes and dendritic cells. Costimulation of effector T cells with anti-CD3 and flagellin resulted in enhanced proliferation and production of IL-2, at levels equivalent to those achieved by costimulation with CD28. In contrast, costimulation with flagellin did not break the hyporesponsiveness of CD4+CD25+ Treg cells, but rather, potently increased their suppressive capacity and enhanced expression of FOXP3. These observations suggest that, in addition to their APC-mediated indirect effects, TLR ligands have the capacity to directly regulate T cell responses and modulate the suppressive activity of Treg cells. PMID- 16339543 TI - Elevated T regulatory cells in long-term stable transplant tolerance in rhesus macaques induced by anti-CD3 immunotoxin and deoxyspergualin. AB - Regulatory T cells (Tregs) are implicated in immune tolerance and are variably dependent on IL-10 for in vivo function. Brief peritransplant treatment of multiple nonhuman primates (NHP) with anti-CD3 immunotoxin and deoxyspergualin has induced stable (5-10 years) rejection-free tolerance to MHC-mismatched allografts, which associated with sustained elevations in serum IL-10. In this study, we demonstrate that resting and activated PBMC from long-term tolerant NHP recipients are biased to secrete high levels of IL-10, compared with normal NHP PBMC. Although IL-10-producing CD4+ Tregs (type 1 regulatory cells (TR1)/IL-10 Tregs) were undetectable (<0.5%) in normal rhesus monkeys, 7.5 +/- 1.7% of circulating CD4+ T cells of tolerant rhesus recipients expressed IL-10. In addition to this >15-fold increase in Tr1/IL-10 Tregs, the tolerant monkeys exhibited a nearly 3-fold increase in CD4+CD25+ Tregs, 8.1 +/- 3.0% of CD4 T cells vs 2.8 +/- 1.4% in normal cohorts (p < 0.02). The frequency of CD4+CD25+IL 10+ cells was elevated 5-fold in tolerant vs normal NHP (1.8 +/- 0.9% vs 0.4 +/- 0.2%). Rhesus CD4+CD25+ Tregs exhibited a memory phenotype, and expressed high levels of Foxp3 and CTLA-4 compared with CD4+CD25- T cells. Also, NHP CD4+CD25+ Tregs proliferated poorly after activation and suppressed proliferation of CD4+CD25- effector T cells, exhibiting regulatory properties similar to rodent and human CD4+CD25+ Tregs. Of note, depletion of CD4+CD25+ Tregs restored indirect pathway antidonor responses in tolerant NHP. Our study demonstrates an expanded presence of Treg populations in tolerant NHP recipients, suggesting that these adaptations may be involved in maintenance of stable tolerance. PMID- 16339544 TI - Erythromycin differentially inhibits lipopolysaccharide- or poly(I:C)-induced but not peptidoglycan-induced activation of human monocyte-derived dendritic cells. AB - Erythromycin (EM) has attracted attention because of its anti-inflammatory effect. Because dendritic cells (DCs) are the most potent APCs involved in numerous pathologic processes including innate immunity, we examined effects of EM on the activation of human DCs by pathogen-derived stimuli. Monocyte-derived DCs were pretreated with EM and subsequently stimulated with peptidoglycan, polyriboinosinic:polyribocytidylic acid (poly(I:C)), or LPS. The activation of DCs was assessed by surface molecule expression and cytokine production. To reveal the signaling pathways affected by EM, TLR expression, NF-kappaB, IFN regulatory factor-3, and AP-1 activation were examined. EM inhibited costimulatory molecule expression and cytokine production that was induced by poly(I:C) and LPS but not by peptidoglycan. EM pretreatment down- and up regulated mRNA levels of TLR3 and TLR2, respectively, but did not affect that of TLR4. EM suppressed IFN regulatory factor-3 activation and IFN-beta production but not AP-1 activation induced by poly(I:C) and LPS. The inhibitory effect of EM on NF-kappaB activation was observed only in poly(I:C)-stimulated DCs. EM selectively suppressed activation of DCs induced by LPS and poly(I:C) in different ways, suggesting that the immuno-modulating effects of EM depend on the nature of pathogens. These results might explain why EM prevents the virus induced exacerbation in the chronic inflammatory respiratory diseases and give us the clue to design new drugs to treat these diseases. PMID- 16339545 TI - Disruption of lipid rafts stimulates phospholipase d activity in human lymphocytes: implication in the regulation of immune function. AB - Recent evidence suggests that phospholipase D (PLD) can be regulated through its association/dissociation to lipid rafts. We show here that modifying lipid rafts either by cholesterol depletion using methyl-beta-cyclodextrin and filipin or by conversion of sphingomyelin to ceramide with exogenous bacterial sphingomyelinase (bSMase) markedly activated the PLD of human PBMC. bSMase was the most potent PLD activator, giving maximal 6- to 7-fold increase in PLD activity. Triton X-100 treated lysates prepared from control PBMC and from bSMase-treated cells were fractionated by centrifugation on sucrose density gradient. We observed that bSMase treatment of the cells induced a larger ceramide increase in raft than in nonraft membranes and displaced both the Src kinase Lck and PLD1 out of the raft fractions. In addition, the three raft-modifying agents markedly inhibited the lymphoproliferative response to mitogenic lectin. To examine further the potential role of PLD activation in the control of lymphocyte responses, we transiently overexpressed either of the PLD1 and PLD2 isoforms in Jurkat cells and analyzed the phorbol ester plus ionomycin-induced expression of IL-2 mRNA, which is one of the early responses of lymphocyte to activation. We observed a 43% decrease of IL-2 mRNA level in Jurkat cells overexpressing PLD1 as compared with mock- or PLD2-transfected cells, which indicates that elevated PLD1, but not PLD2, activity impairs lymphocyte activation. Altogether, the present results support the hypothesis that PLD1 is activated by exclusion from lipid rafts and that this activation conveys antiproliferative signals in lymphoid cells. PMID- 16339546 TI - Identification of immunodominant epitopes of alpha-gliadin in HLA-DQ8 transgenic mice following oral immunization. AB - Celiac disease, triggered by wheat gliadin and related prolamins from barley and rye, is characterized by a strong association with HLA-DQ2 and HLA-DQ8 genes. Gliadin is a mixture of many proteins that makes difficult the identification of major immunodominant epitopes. To address this issue, we expressed in Escherichia coli a recombinant alpha-gliadin (r-alpha-gliadin) showing the most conserved sequence among the fraction of alpha-gliadins. HLA-DQ8 mice, on a gluten-free diet, were intragastrically immunized with a chymotryptic digest of r-alpha gliadin along with cholera toxin as adjuvant. Spleen and mesenteric lymph node T cell responses were analyzed for in vitro proliferative assay using a panel of synthetic peptides encompassing the entire sequence of r-alpha-gliadin. Two immunodominant epitopes corresponding to peptide p13 (aa 120-139) and p23 (aa 220 239) were identified. The response was restricted to DQ and mediated by CD4+ T cells. In vitro tissue transglutaminase deamidation of both peptides did not increase the response; furthermore, tissue transglutaminase catalyzed extensive deamidation in vitro along the entire r-alpha-gliadin molecule, but failed to elicit new immunogenic determinants. Surprisingly, the analysis of the cytokine profile showed that both deamidated and native peptides induced preferentially IFN-gamma secretion, despite the use of cholera toxin, a mucosal adjuvant that normally induces a Th2 response to bystander Ags. Taken together, these data suggest that, in this model of gluten hypersensitivity, deamidation is not a prerequisite for the initiation of gluten responses. PMID- 16339547 TI - A role for TLRs in the regulation of immune cell migration by first trimester trophoblast cells. AB - Normal pregnancy is characterized by the presence of innate immune cells at the maternal-fetal interface. Originally, it was postulated that the presence of these leukocytes was due to an immune response toward paternal Ags expressed by the invading trophoblasts. Instead, we and others postulate that these innate immune cells are necessary for successful implantation and pregnancy. However, elevated leukocyte infiltration may be an underlying cause of pregnancy complications, such as preterm labor or preeclampsia. Furthermore, such conditions have been attributed to an intrauterine infection. Therefore, we hypothesize that first trimester trophoblast cells, upon recognition of microbes through TLRs, may coordinate an immune response by recruiting cells of the innate immune system to the maternal-fetal interface. In this study, we have demonstrated that human first trimester trophoblast cells constitutively secrete the chemokines growth-related oncogene, growth-related oncogene alpha, IL-8, and MCP-1 and are able to recruit monocytes and NK cells, and to a lesser degree, neutrophils. Following the ligation of TLR-3 by the viral ligand, poly(I:C), or TLR-4 by bacterial LPS, trophoblast secretion of chemokines is significantly increased and this in turn results in elevated monocyte and neutrophil chemotaxis. In addition, TLR-3 stimulation also induces trophoblast cells to secrete RANTES. These results suggest a novel mechanism by which first trimester trophoblast cells may differentially modulate the maternal immune system during normal pregnancy and in the presence of an intrauterine infection. Such altered trophoblast cell responses might contribute to the pathogenesis of certain pregnancy complications. PMID- 16339548 TI - Somatic hypermutation and junctional diversification at Ig heavy chain loci in the nurse shark. AB - We estimate there are approximately 15 IgM H chain loci in the nurse shark genome and have characterized one locus. It consists of one V, two D, and one J germline gene segments, and the constant (C) region can be distinguished from all of the others by a unique combination of restriction endonuclease sites in Cmu2. On the basis of these Cmu2 markers, 22 cDNA clones were selected from an epigonal organ cDNA library from the same individual; their C region sequences proved to be the same up to the polyadenylation site. With the identification of the corresponding germline gene segments, CDR3 from shark H chain rearrangements could be analyzed precisely, for the first time. Considerable diversity was generated by trimming and N addition at the three junctions and by varied recombination patterns of the two D gene segments. The cDNA sequences originated from independent rearrangements events, and most carried both single and contiguous substitutions. The 53 point mutations occurred with a bias for transition changes (53%), whereas the 78 tandem substitutions, mostly 2-4 bp long, do not (36%). The nature of the substitution patterns is the same as for mutants from six loci of two nurse shark L chain isotypes, showing that somatic hypermutation events are very similar at both H and L chain genes in this early vertebrate. The cis-regulatory elements targeting somatic hypermutation must have already existed in the ancestral Ig gene, before H and L chain divergence. PMID- 16339549 TI - Specific IgE determination to epitope peptides of omega-5 gliadin and high molecular weight glutenin subunit is a useful tool for diagnosis of wheat dependent exercise-induced anaphylaxis. AB - Wheat omega-5 gliadin and a high m.w. glutenin subunit (HMW-glutenin) have been reported as major allergens in wheat-dependent exercise-induced anaphylaxis. A simultaneous detection of specific IgE to epitope sequences of both proteins is considered to be a reliable method for diagnosis of wheat-dependent exercise induced anaphylaxis. However, the IgE-binding epitope of HMW-glutenin remains unknown. The aim of this study was to determine the IgE-binding epitopes of HMW glutenin to establish a useful system of identifying patients with wheat dependent exercise-induced anaphylaxis. For determination of IgE-binding epitopes of HMW-glutenin overlapping peptides were synthesized and reactivities of IgE Abs in the sera of patients to those peptides were analyzed. Three IgE-binding epitopes, QQPGQ, QQPGQGQQ, and QQSGQGQ, were identified within primary sequence of HMW-glutenin. Epitope peptides, which include IgE-binding sequences of omega-5 gliadin and a HMW-glutenin, were synthesized and peptide-specific IgE Abs were measured by CAP-System fluorescent enzyme immunoassay. Twenty-nine of 30 patients with wheat-dependent exercise-induced anaphylaxis had specific IgE Abs to these epitope peptides. None of the 25 sera from healthy subjects reacted to both epitope peptides. Twenty-five patients with atopic dermatitis who had specific IgE to wheat and/or gluten had very low or nonexistent levels of epitope peptide specific IgE Abs. These results indicated that measurement of IgE levels specific to epitope peptides of omega-5 gliadin and HMW-glutenin is useful as an in vitro diagnostic method for the assessment of patients with wheat-dependent exercise induced anaphylaxis. PMID- 16339550 TI - T cell activation-induced CrkII binding to the Zap70 protein tyrosine kinase is mediated by Lck-dependent phosphorylation of Zap70 tyrosine 315. AB - The Zap70 protein tyrosine kinase controls TCR-linked signal transduction pathways and is critical for T cell development and responsiveness. Following engagement of TCR, the Zap70 undergoes phosphorylation on multiple tyrosine residues that are implicated in the regulation of its catalytic activity and interaction with signaling effector molecules downstream of the TCR. We have shown previously that the CT10 regulator of kinase II (CrkII) adapter protein interacts with tyrosine-phosphorylated Zap70 in TCR-engaged T cells, and now extend these studies to show that Tyr315 in the Zap70 interdomain B region is the site of interaction with CrkII. A point mutation of Tyr315 (Y315F) eliminated the CrkII-Zap70 interaction capacity. Phosphorylation of Tyr315 and Zap70 association with CrkII were both dependent upon the Lck protein tyrosine kinase. Previous studies demonstrated the Tyr315 is the Vav-Src homology 2 (SH2) binding site, and that replacement of Tyr315 by Phe impaired the function of Zap70 in TCR signaling. However, fluorescence polarization-based binding studies revealed that the CrkII-SH2 and the Vav-SH2 bind a phosphorylated Tyr315-Zap70-derived peptide with affinities of a similar order of magnitude (Kd of 2.5 and 1.02 microM, respectively). The results suggest therefore that the biological functions attributed to the association of Zap70 with Vav following T cell activation may equally reflect the association of Zap70 with CrkII, and further support a regulatory role for CrkII in the TCR-linked signal transduction pathway. PMID- 16339551 TI - Sequence transfers between variable regions in a mouse antibody transgene can occur by gene conversion. AB - Different vertebrate species show widely differing usage of somatic hyperconversion (SHC) as a mechanism for diversifying expressed Ab V genes. The basis for the differing levels of SHC in different species is not known. Although no clear evidence for SHC has been found in normal mouse B cells, transgenic mice carrying high-copy numbers of a gene construct designed to optimize detection of SHC have previously been shown to exhibit sequence transfers that resemble gene conversion events. However, these transgene sequence transfers could reflect multistep or reciprocal DNA recombination events rather than gene conversions. We now find in low-copy number transgenic mice that transgene sequence transfers can exhibit the unidirectional sequence information movement that is a hallmark of gene conversion. This indicates that gene conversion between V region sequences can occur in mouse B cells; we propose that the lack of efficient SHC contributions to Ab diversification in normal mice may be due, at least in part, to the particular pattern of V gene recombinational accessibility that occurs in differentiating mouse B cells. PMID- 16339552 TI - A role for phosphatidylinositol 3-kinase in TCR-stimulated ERK activation leading to paxillin phosphorylation and CTL degranulation. AB - PI3K is an important regulator of a number of cellular processes. We examined the contribution of PI3K to mouse CTL signaling, leading to degranulation. We show that TCR-triggered, but not phorbol ester and calcium ionophore-induced, CTL degranulation is dependent on PI3K activity. Although PI3K activity is required for optimal LFA-1-mediated adhesion and cell spreading, this most likely does not account for its full contribution to degranulation. We demonstrate that PI3K is required for TCR-stimulated ERK activation in CTL, which we have shown previously to be required for CTL degranulation. We thus define a pathway through which PI3K most likely regulates degranulation and in which ERK appears to be a key signaling molecule. Furthermore, we identified the cytoskeletal adaptor paxillin as a target of ERK downstream of TCR stimulation. Consistent with a role in degranulation, we demonstrate that paxillin is localized to the microtubule organizing center in resting cells and upon target cell binding is recruited to the contact point with the target cell. These studies demonstrate that PI3K regulates ERK activity leading to CTL degranulation, and identify paxillin as a target of ERK downstream of the TCR. That paxillin is independently phosphorylated by both tyrosine kinase(s) and ERK downstream of the TCR and localized both at the microtubule organizing center and at the target cell contact point suggests an important role for paxillin in CTL-mediated killing. PMID- 16339553 TI - Intergenic transcription is not required in Th2 cells to maintain histone acetylation and transcriptional permissiveness at the Il4-Il13 locus. AB - Noncoding RNA transcripts mapping to intergenic regions of the Il4-Il13 locus have been detected in Th2 cells harboring transcriptionally permissive Il4 and Il13 genes but not in Th1 cells where these genes are repressed. This correlation has given rise to the idea that intergenic transcription may be involved in maintaining the "open" chromatin structure of the Il4-Il13 locus in Th2 cells. We present evidence from real-time RT-PCR, nuclear run on, chromatin immunoprecipitation and 5,6-dichlorobenzimidazole 1-beta-D-ribofuranoside mediated transcriptional inhibition analyses that argue against this hypothesis. Instead, our results are consistent with an alternative role for intergenic transcription in the maintenance of transcriptional silence in Th1-primed cells. PMID- 16339554 TI - Functional interplay between intrinsic B and T cell defects leads to amplification of autoimmune disease in New Zealand black chromosome 1 congenic mice. AB - Genetic loci on New Zealand Black (NZB) chromosome 1 play an important role in the development of lupus-like autoimmune disease. We have shown previously that C57BL/6 mice with an introgressed NZB chromosome 1 interval extending from approximately 35 to 106 cM have significantly more severe autoimmunity than mice with a shorter interval extending from approximately 82 to 106 cM. Comparison of the cellular phenotype in these mice revealed that both mouse strains had evidence of increased T cell activation; however, activation was more pronounced in mice with the longer interval. Mice with the longer interval also had increased B cell activation, leading us to hypothesize that there were at least two independent lupus susceptibility loci on chromosome 1. In this study, we have used mixed hemopoietic radiation chimeras to demonstrate that autoimmunity in these mice arises from intrinsic B and T cell functional defects. We further show that a T cell defect, localized to the shorter interval, leads to spontaneous activation of T cells specific for nucleosome histone components. Despite activation of self-reactive T cells in mixed chimeric mice, only chromosome 1 congenic B cells produce anti-nuclear Abs and undergo class switching, indicating impaired B cell tolerance mechanisms. In mice with the longer chromosome 1 interval, an additional susceptibility locus exacerbates autoimmune disease by producing a positive feedback loop between T and B cell activation. Thus, T and B cell defects act in concert to produce and amplify the autoimmune phenotype. PMID- 16339555 TI - Mice deficient in LRG-47 display enhanced susceptibility to Trypanosoma cruzi infection associated with defective hemopoiesis and intracellular control of parasite growth. AB - IFN-gamma is known to be required for host control of intracellular Trypanosoma cruzi infection in mice, although the basis of its protective function is poorly understood. LRG-47 is an IFN-inducible p47GTPase that has been shown to regulate host resistance to intracellular pathogens. To investigate the possible role of LRG-47 in IFN-gamma-dependent control of T. cruzi infection, LRG-47 knockout (KO) and wild-type (WT) mice were infected with the Y strain of this parasite, and host responses were analyzed. When assayed on day 12 after parasite inoculation, LRG-47 KO mice, in contrast to IFN-gamma KO mice, controlled early parasitemia almost as effectively as WT animals. However, the infected LRG-47 KO mice displayed a rebound in parasite growth on day 15, and all succumbed to the infection by day 19. Additional analysis indicated that LRG-47-deficient mice exhibit unimpaired proinflammatory responses throughout the infection. Instead, reactivated disease in the KO animals was associated with severe splenic and thymic atrophy, anemia, and thrombocytopenia not observed in their WT counterparts. In addition, in vitro studies revealed that IFN-gamma-stimulated LRG-47 KO macrophages display defective intracellular killing of amastigotes despite normal expression of TNF and NO synthetase type 2 and that both NO synthetase type 2 and LRG-47 are required for optimum IFN-gamma-dependent restriction of parasite growth. Together, these data establish that LRG-47 can influence pathogen control by simultaneously regulating macrophage-microbicidal activity and hemopoietic function. PMID- 16339556 TI - Defective microarchitecture of the spleen marginal zone and impaired response to a thymus-independent type 2 antigen in mice lacking scavenger receptors MARCO and SR-A. AB - The macrophage scavenger receptor macrophage receptor with a collagenous structure (MARCO) is expressed in mice by the marginal zone macrophages of the spleen and by macrophages of the medullary cords of lymph nodes, as well as the peritoneal macrophages. MARCO is a relative of scavenger receptor A (SR-A), the more widely expressed prototypic member of the scavenger receptor family. In the present study, we found that genetic ablation of MARCO leads to changes in the organization of the splenic marginal zone, and causes a significant reduction in the size of the resident peritoneal macrophage population, possibly due to changes in adhesion and migration capacity. In mice lacking both MARCO and SR-A these effects are even more apparent. During ontogeny, the appearance and organization of the MARCO-expressing cells in the spleen precedes the appearance of other receptors on macrophages in the marginal zone, such as SIGNR1 and Siglec 1. In the absence of MARCO, a clear delay in the organization of the marginal zone was observed. Similar findings were seen when the reappearance of the various subsets from precursors was studied after depleting macrophages from the adult spleen by a liposome treatment. When challenged with a pneumococcal polysaccharide vaccine, a T-independent type 2 Ag for which an intact marginal zone is crucial, the knockout mice exhibited a clearly impaired response. These findings suggest that both MARCO and SR-A, in addition to being important scavenger receptors, could be involved in the positioning and differentiation of macrophages, possibly through interaction with endogenous ligands. PMID- 16339557 TI - Activated macrophages infected with Legionella inhibit T cells by means of MyD88 dependent production of prostaglandins. AB - To understand how macrophages (Mphi) activated with IFN-gamma modulate the adaptive immune response to intracellular pathogens, the interaction of IFN-gamma treated bone marrow-derived murine Mphi (BMphi) with Legionella pneumophila was investigated. Although Legionella was able to evade phagosome lysosome fusion initially, and was capable of de novo protein synthesis within IFN-gamma-treated BMphi, intracellular growth of Legionella was restricted. It was determined that activated BMphi infected with Legionella suppressed IFN-gamma production by Ag specific CD4 and CD8 T cells. A factor sufficient for suppression of T cell responses was present in culture supernatants isolated from activated BMphi following Legionella infection. Signaling pathways requiring MyD88 and TLR2 were important for production of a factor produced by IFN-gamma-treated BMphi that interfered with effector T cell functions. Cyclooxygenase-2-dependent production of PGs by IFN-gamma-treated BMphi infected with Legionella was required for inhibition of effector T cell responses. From these data we conclude that activated Mphi can down-modulate Ag-specific T cell responses after they encounter bacterial pathogens through production of PGs, which may be important in preventing unnecessary immune-mediated damage to host tissues. PMID- 16339558 TI - A requirement for the Vgamma1+ subset of peripheral gammadelta T cells in the control of the systemic growth of Toxoplasma gondii and infection-induced pathology. AB - gammadelta T cells are a diverse population of T cells that are widely distributed and are a common feature of pathogen-induced immune responses. It is not clear, however, whether different populations of gammadelta T cells have specific functions, and what factors determine the functional properties of individual populations. A murine model of peroral Toxoplasma gondii infection was used to determine the contribution Vgamma1+ intestinal intraepithelial lymphocytes (IELs) vs systemic Vgamma1+ T cells make to the acute and chronic stages of the host immune response, and whether the macrophage cytocidal activity of Vgamma1+ T cells described in bacterial infections is seen in other, unrelated infectious disease models. In response to oral infection with virulent type 1 or avirulent type II strains of T. gondii, TCR-delta-/- mice rapidly developed severe ileitis. In contrast, in mice deficient in Vgamma1+ T cells and IELs and wild-type mice, inflammation was delayed in onset and less severe. The protective effect of (Vgamma1-) IELs to Toxoplasma infection was unrelated to their cytolytic and cytokine (Th1)-producing capabilities. Systemic Vgamma1+ T cells were shown to play an essential role in limiting parasite growth and inflammation in peripheral tissues and, in particular, in the CNS, that was associated with their ability to efficiently kill parasite-elicited and infected macrophages. These findings suggest that macrophage cytocidal activity of Vgamma1+ T cells may be a universal feature of pathogen-induced immune responses and that microenvironmental factors influence the involvement and function of gammadelta T cells in the host response to infection. PMID- 16339559 TI - CD11b+/Gr-1+ immature myeloid cells mediate suppression of T cells in mice bearing tumors of IL-1beta-secreting cells. AB - Tumor cells secreting IL-1beta are invasive and metastatic, more than the parental line or control mock-transfected cells, and concomitantly induce in mice general immune suppression of T cell responses. Suppression strongly correlates with accumulation in the peripheral blood and spleen of CD11b+/Gr-1+ immature myeloid cells and hematological alterations, such as splenomegaly, leukocytosis, and anemia. Resection of large tumors of IL-1beta-secreting cells restored immune reactivity and hematological alterations within 7-10 days. Treatment of tumor bearing mice with the physiological inhibitor of IL-1, the IL-1R antagonist, reduced tumor growth and attenuated the hematological alterations. Depletion of CD11b+/Gr-1+ immature myeloid cells from splenocytes of tumor-bearing mice abrogated suppression. Despite tumor-mediated suppression, resection of large tumors of IL-1beta-secreting cells, followed by a challenge with the wild-type parental cells, induced resistance in mice; protection was not observed in mice bearing tumors of mock-transfected fibrosarcoma cells. Altogether, we show in this study that tumor-derived IL-1beta, in addition to its proinflammatory effects on tumor invasiveness, induces in the host hematological alterations and tumor-mediated suppression. Furthermore, the antitumor effectiveness of the IL-1R antagonist was also shown to encompass restoration of hematological alterations, in addition to its favorable effects on tumor invasiveness and angiogenesis that have previously been described by us. PMID- 16339560 TI - Yersinia outer protein P suppresses TGF-beta-activated kinase-1 activity to impair innate immune signaling in Yersinia enterocolitica-infected cells. AB - Pathogenic Yersinia spp. use a panel of virulence proteins that antagonize signal transduction processes in infected cells to undermine host defense mechanisms. One of these proteins, Yersinia enterocolitica outer protein P (YopP), down regulates the NF-kappaB and MAPK signaling pathways, which suppresses the proinflammatory host immune response. In this study, we explored the mechanism by which YopP succeeds to simultaneously disrupt several of these key signaling pathways of innate immunity. Our data show that YopP operates upstream of its characterized eukaryotic binding partner IkappaB kinase-beta to shut down the NF kappaB signaling cascade. Accordingly, YopP efficiently impaired the activities of TGF-beta-activated kinase-1 (TAK1) in infected cells. TAK1 is an important activator of the IkappaB kinase complex in the TLR signaling cascade. The repression of TAK1 activities correlated with reduced activation of NF-kappaB- as well as AP-1-dependent reporter gene expression in Yersinia-infected murine macrophages. This suggests that the impairment of the TAK1 enzymatic activities by Yersinia critically contributes to down-regulate activation of NF-kappaB and of MAPK members in infected host cells. The inhibition of TAK1 potentially results from the blockade of signaling events that control TAK1 induction. This process could involve the attenuation of ubiquitination of the upstream signal transmitter TNFR-associated factor-6. Together, these results indicate that, by silencing the TAK1 signaling complex, Yersinia counteracts the induction of several conserved signaling pathways of innate immunity, which aids the bacterium in subverting the host immune response. PMID- 16339561 TI - Cytomegalovirus-specific CD4+ T cells in healthy carriers are continuously driven to replicative exhaustion. AB - Repeated antigenic encounter drives proliferation and differentiation of memory T cell pools. An important question is whether certain specific T cells may be driven eventually to exhaustion in elderly individuals since the human life expectancy is increasing. We found that CMV-specific CD4+ T cells were significantly expanded in healthy young and old carriers compared with purified protein derivative-, varicella zoster virus-, EBV-, and HSV-specific populations. These CMV-specific CD4+ T cells exhibited a late differentiated phenotype since they were largely CD27 and CD28 negative and had shorter telomeres. Interestingly, in elderly CMV-seropositive subjects, CD4+ T cells of different specificities were significantly more differentiated than the same cells in CMV seronegative individuals. This suggested the involvement of bystander-secreted, differentiation-inducing factors during CMV infection. One candidate was IFN alpha, which induced loss of costimulatory receptors and inhibited telomerase in activated CD4+ T cells and was secreted at high levels by CMV-stimulated plasmacytoid dendritic cells (PDC). The CMV-specific CD4+ T cells in elderly subjects had severely restricted replicative capacity. This is the first description of a human memory T cell population that is susceptible to being lost through end-stage differentiation due to the combined effects of lifelong virus reactivation in the presence of bystander differentiation-inducing factors. PMID- 16339562 TI - E2 of hepatitis C virus inhibits apoptosis. AB - Hepatitis C virus (HCV) is the major causative agent of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma, and can be involved in very long chronic infections up to 30 years or more. Therefore, it has been speculated that HCV possesses mechanisms capable of modulating host defense systems such as innate and adaptive immunity. To investigate this virus-host interaction, we generated HCV replicons containing various HCV structural proteins and then analyzed the sensitivity of replicon-containing cells to the apoptosis-inducing agent, TRAIL. TRAIL-induced apoptosis was monitored by cleavage of procaspase-3 and procaspase 9 as well as that of their substrate poly(ADP-ribose) polymerase. TRAIL-induced apoptosis was inhibited in cells expressing HCV E2. Moreover, expression of HCV E2 enhanced the colony forming efficiency of replicon-containing cells by 25 fold. Blockage of apoptosis by E2 seems to be related to inhibition of TRAIL induced cytochrome c release from the mitochondria. Based on these results, we propose that E2 augments persistent HCV infection by blocking host-induced apoptosis of infected cells. PMID- 16339563 TI - Pure lipopolysaccharide or synthetic lipid A induces activation of p21Ras in primary macrophages through a pathway dependent on Src family kinases and PI3K. AB - Recognition of bacterial LPS by macrophages plays a critical role in host defense against infection by Gram-negative bacteria. However, when not tightly regulated, the macrophage's response to LPS can induce severe disease and septic shock. Although LPS triggers the activation of multiple signaling pathways in macrophages, it was unclear whether these include activation of the p21Ras GTPases. We report that p21Ras is rapidly and transiently activated in murine primary macrophages stimulated with an ultra-pure preparation of LPS or with synthetic lipid A. The molecular basis of this activation was investigated using a pharmacological approach. LPS-induced activation of p21Ras was inhibited in the presence of PP2, LY294002, or wortmannin, suggesting that it depends on the activity of one or more members of the Src kinase family and the subsequent activation of PI3K. In that pharmacological inhibitors of PI3K inhibited LPS induced activation of p21Ras, but not activation of ERK, we concluded that LPS induced activation of ERK occurs through a pathway that is not dependent on the activation of p21Ras. PMID- 16339564 TI - Helicobacter pylori-Induced invasion and angiogenesis of gastric cells is mediated by cyclooxygenase-2 induction through TLR2/TLR9 and promoter regulation. AB - Cyclooxygenase-2 (COX-2) plays a crucial role in Helicobacter pylori-associated gastric cancer. In this study, we report that H. pylori-induced COX-2 expression enhances the cancer cell invasion and angiogenesis via TLR2 and TLR9, which can be attenuated by the specific COX-2 inhibitor NS398 or celecoxib. The cAMP response element (CRE) and AP1 sites, but not kappaB on the COX-2 promoter, are involved in MAPKs-regulated COX-2 expression. Differential bindings of the CREB 1, ATF-2, c-jun to the CRE site, and the c-fos, c-jun, ATF-2 to the AP1 site are demonstrated by DNA affinity protein-binding, supershift, and chromatin immunoprecipitation assays. Activations of these transcription factors were attenuated by different MAPKs inhibitors. The mutants of TLR2, TLR9, or MAPKs inhibited H. pylori-induced COX-2 promoter, CRE, and AP-1 activities. MAPKs inhibitors attenuated the H. pylori-induced COX-2 mRNA and protein expressions. These results indicate that H. pylori acts through TLR2 and TLR9 to activate MAPKs, especially p38, and their downstream transcription factors (CREB-1, ATF-2, c-jun, and c-fos), resulting in the activations of CRE and AP-1 on the COX-2 promoter. These intracellular networks drive the COX-2-dependent PGE2 release and contribute to cell invasion and angiogenesis. PMID- 16339565 TI - Allergen-induced airway hyperresponsiveness mediated by cyclooxygenase inhibition is not dependent on 5-lipoxygenase or IL-5, but is IL-13 dependent. AB - Cyclooxygenase (COX) inhibition during allergic sensitization and allergen airway challenge results in augmented allergic inflammation. We hypothesized that this increase in allergic inflammation was dependent on increased generation of leukotrienes that results from COX inhibition, as leukotrienes are important proinflammatory mediators of allergic disease. To test this hypothesis, we allergically sensitized and challenged mice deficient in 5-lipoxygenase (5-LO). We found that 5-LO knockout mice that were treated with a COX inhibitor during allergic sensitization and challenge had significantly increased airway hyperresponsiveness (AHR) (p < 0.01) and airway eosinophilia (p < 0.01) compared with 5-LO knockout mice that were treated with vehicle. The proinflammatory cytokines have also been hypothesized to be critical regulators of airway inflammation and AHR. We found that the increase in airway eosinophilia seen with COX inhibition is dependent on IL-5, whereas the increase in AHR is not dependent on this cytokine. In contrast, the COX inhibition-mediated increase in AHR is dependent on IL-13, but airway eosinophilia is not. These results elucidate the pathways by which COX inhibition exerts a critical effect of the pulmonary allergen-induced inflammatory response and confirm that COX products are important regulators of allergic inflammation. PMID- 16339567 TI - CD8 T cell-mediated lung damage in response to the extracellular pathogen pneumocystis is dependent on MHC class I expression by radiation-resistant lung cells. AB - Pneumocystis, a fungal, extracellular pathogen causes a life-threatening pneumonia in patients with severe immunodeficiencies. In the absence of CD4 T cells, Pneumocystis infection results in vigorous CD8 T cell influx into the alveolar and interstitial spaces of the lung. This response results in lung damage characterized by low pO2 and albumin leakage into the bronchoalveolar lavage fluid similar to other CD8 T cell-mediated interstitial lung diseases. How this extracellular pathogen elicits a CD8 T cell response is not clear, and it was the aim of our study to determine the Ag specificity of the recruited CD8 T cells and to determine whether MHC class I (MHC I) expression was necessary to initiate lung damage. Using an adoptive T cell-transfer model with either polyclonal wild-type CD8 T cells or transgenic influenza virus-specific CD8 T cells we found that CD8 T cell recruitment is Ag-specific and requires the continuous presence of the Pneumocystis pathogen. Bone marrow chimera experiments using Rag-1 and beta2-microglobulin-deficient mice as hosts demonstrated a requirement for MHC I expression on nonbone marrow-derived cells of the lung. This suggests either direct processing of Pneumocystis Ags by nonbone marrow derived cells of the lung or the induction of lung damage triggered by a lung specific autoantigen. Using perforin-, Fas-, and IFN-gamma-deficient animals, we showed that these molecules are not directly involved in the CD8-mediated lung damage. However, CD8 T cell-mediated lung damage is Ag-specific is induced by a MHC I-expressing nonbone marrow-derived cell in the lung and is dependent on the continued presence of live Pneumocystis. PMID- 16339566 TI - Adenosine augments IL-10 production by macrophages through an A2B receptor mediated posttranscriptional mechanism. AB - Adenosine receptor ligands have anti-inflammatory effects and modulate immune responses by up-regulating IL-10 production by immunostimulated macrophages. The adenosine receptor family comprises G protein-coupled heptahelical transmembrane receptors classified into four types: A1, A2A, A2B, and A3. Our understanding of the signaling mechanisms leading to enhanced IL-10 production following adenosine receptor occupancy on macrophages is limited. In this study, we demonstrate that adenosine receptor occupancy increases IL-10 production by LPS-stimulated macrophages without affecting IL-10 promoter activity and IL-10 mRNA levels, indicating a posttranscriptional mechanism. Transfection experiments with reporter constructs containing sequences corresponding to the AU-rich 3' untranslated region (UTR) of IL-10 mRNA confirmed that adenosine receptor activation acts by relieving the translational repressive effect of the IL-10 3' UTR. By contrast, adenosine receptor activation failed to liberate the translational arrest conferred by the 3'-UTR of TNF-alpha mRNA. The IL-10 3'-UTR formed specific complexes with proteins present in cytoplasmic extracts of RAW 264.7 cells. Adenosine enhanced binding of proteins to a region of the IL-10 3' UTR containing the GUAUUUAUU nonamer. The stimulatory effect of adenosine on IL 10 production was mediated through the A(2B) receptor, because the order of potency of selective agonists was 5'-N-ethylcarboxamidoadenosine (NECA) > N6-(3 iodobenzyl)-adenosine-5'-N-methyluronamide (IB-MECA) > 2-chloro-N6 cyclopentyladenosine (CCPA) = 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethyl carboxamidoadenosine (CGS-21680). Also, the selective A2B antagonist, alloxazine, prevented the effect of adenosine. Collectively, these studies identify a novel pathway in which activation of a G protein-coupled receptor augments translation of an anti-inflammatory gene. PMID- 16339568 TI - IL-18 induces monocyte chemotactic protein-1 production in macrophages through the phosphatidylinositol 3-kinase/Akt and MEK/ERK1/2 pathways. AB - Macrophages are activated during an inflammatory response and produce multiple inflammatory cytokines. IL-18 is one of the most important innate cytokines produced from macrophages in the early stages of the inflammatory immune response. Monocyte chemoattractant protein (MCP-1) is expressed in many inflammatory diseases such as multiple sclerosis and rheumatoid arthritis, and its expression is correlated with the severity of the disease. Both IL-18 and MCP 1 have been shown to be involved in inflammatory immune responses. However, it has been unclear whether IL-18 is involved in the induction of MCP-1. This investigation was initiated to determine whether IL-18 can induce MCP-1 production, and if so, by which signal transduction pathways. We found that IL-18 induced the production of MCP-1 in macrophages, which was IL-12-independent and was not mediated by autocrine cytokines such as IFN-gamma or TNF-alpha. We then examined signal transduction pathways involved in IL-18-induced MCP-1 production. We found that IL-18 did not activate the IkappaB kinase/NF-kappaB pathway, evidenced by no degradation of IkappaBalpha and no translocation of NF-kappaB p65 to the nucleus in IL-18-stimulated macrophages. Instead, IL-18 activated the PI3K/Akt and MEK/ERK1/2 pathways. Inhibition of either of these pathways attenuated MCP-1 production in macrophages, and inhibition of both signaling pathways resulted in the complete inhibition of MCP-1 production. On the basis of these observations, we conclude that IL-18 induces MCP-1 production through the PI3K/Akt and MEK/ERK1/2 pathways in macrophages. PMID- 16339569 TI - Differential regulation of osteoblast activity by Th cell subsets mediated by parathyroid hormone and IFN-gamma. AB - Bone loss is a typical pathological feature of chronic inflammatory bone diseases including rheumatoid arthritis, in which CD4 effector T cells play critical roles. We found that activated mouse Th2 and not Th1 cells produced the parathyroid hormone (PTH). Unlike in the parathyroid cells, PTH expression in Th2 cells was not regulated by the fluctuation of calcium level, but rather it required the full activation of the T cells. Although PTH was expressed in immature Th2 cells, and its receptor was transiently expressed during Th1 and Th2 cell differentiation, PTH did not significantly affect the outcome of the differentiation. In primary osteoblasts cultured in Th2 cell condition medium, the alkaline phosphatase (ALP) activity was maintained at a basal level. However, antagonizing PTH in the condition medium resulted in a significant reduction of the ALP activity. These results demonstrated an important role of the Th2 cell derived PTH in maintaining the bone-forming activity of the osteoblasts under inflammatory conditions. In osteoblasts cultured in the Th1 cell condition medium, the ALP activity was significantly suppressed. Neutralizing IFN-gamma alleviated the suppression. Conversely, treatment of osteoblasts with IFN-gamma suppressed the ALP activity. Unlike ALP, expression of the major bone matrix proteins by the osteoblasts was only minimally affected by either Th1 or Th2 cytokine environment. In addition, the Th2 cytokine environment also regulated to expression of receptor activator of NF-kappaB ligand and osteoprotegerin through both PTH-dependent and -independent mechanisms. Our study therefore identified new regulatory events in bone remodeling under inflammatory conditions. PMID- 16339571 TI - TNFalpha-induced long pentraxin PTX3 expression in human lung epithelial cells via JNK. AB - Long pentraxin 3 (PTX3), an acute-phase protein, is a newly clarified mediator for innate immunity and inflammation. As a soluble pattern recognition receptor, it has a nonredundant role in antifungal infection. Overexpression of PTX3 worsens acute lung injury. The lung epithelium is a critical factor in defense against pulmonary pathogens; it is also involved in acute inflammatory responses related to tissue injury. However, very little is known about how PTX3 is regulated in the lung epithelium. In this study, we found that i.v. injection of LPS induced PTX3 expression in rat lung alveolar epithelium. Using human lung cell lines and primary epithelial cells, we found that PTX3 expression was significantly up-regulated by TNF-alpha in a time- and dose-dependent manner, but not by LPS. Pretreatment with either actinomycin D or cycloheximide abolished TNF alpha-induced PTX3 expression, indicating the requirement for both transcriptional and translational regulation. The TNF-alpha-induced PTX3 expression was blocked by SP600125, a JNK-specific inhibitor, but not by the inhibitors against NF-kappaB, ERKs, or p38 MAPK. Knockdown of either JNK1 or JNK2 with small interfering RNA also significantly reduced the regulated PTX3 expression. Thus, lung epithelial cells appear to be a major local source for PTX3 production, which could be induced in vivo from these cells by LPS or other inflammatory stimuli, and may be an important mediator for host defense and tissue damage. The importance of the JNK pathway for the regulated PTX3 expression may be a potential target for its regulation in the lung. PMID- 16339570 TI - Inflammation-induced chondrocyte hypertrophy is driven by receptor for advanced glycation end products. AB - The multiligand receptor for advanced glycation end products (RAGE) mediates certain chronic vascular and neurologic degenerative diseases accompanied by low grade inflammation. RAGE ligands include S100/calgranulins, a class of low molecular-mass, calcium-binding polypeptides, several of which are chondrocyte expressed. Here, we tested the hypothesis that S100A11 and RAGE signaling modulate osteoarthritis (OA) pathogenesis by regulating a shift in chondrocyte differentiation to hypertrophy. We analyzed human cartilages and cultured human articular chondrocytes, and used recombinant human S100A11, soluble RAGE, and previously characterized RAGE-specific blocking Abs. Normal human knee cartilages demonstrated constitutive RAGE and S100A11 expression, and RAGE and S100A11 expression were up-regulated in OA cartilages studied by immunohistochemistry. CXCL8 and TNF-alpha induced S100A11 expression and release in cultured chondrocytes. Moreover, S100A11 induced cell size increase and expression of type X collagen consistent with chondrocyte hypertrophy in vitro. CXCL8-induced, IL-8 induced, and TNF-alpha-induced but not retinoic acid-induced chondrocyte hypertrophy were suppressed by treatment with soluble RAGE or RAGE-specific blocking Abs. Last, via transfection of dominant-negative RAGE and dominant negative MAPK kinase 3, we demonstrated that S100A11-induced chondrocyte type X collagen expression was dependent on RAGE-mediated p38 MAPK pathway activation. We conclude that up-regulated chondrocyte expression of the RAGE ligand S100A11 in OA cartilage, and RAGE signaling through the p38 MAPK pathway, promote inflammation-associated chondrocyte hypertrophy. RAGE signaling thereby has the potential to contribute to the progression of OA. PMID- 16339572 TI - Complement-dependent apoptosis and inflammatory gene changes in murine lupus cerebritis. AB - The role of complement activation in the brains of MRL/lpr lupus mice was determined using the potent C3 convertase inhibitor, CR1-related y (Crry), administered both as an overexpressing Crry transgene and as Crry-Ig. Prominent deposition of complement proteins C3 and C9 in brains of MRL/lpr mice was indicative of complement activation and was significantly reduced by Crry. Apoptosis was determined in brain using different independent measures of apoptosis, including TUNEL staining, DNA laddering, and caspase-3 activity, all of which were markedly increased in lupus mice and could be blocked by inhibiting complement with Crry. Complement activation releases inflammatory mediators that can induce apoptosis. The mRNA for potentially proinflammatory proteins such as TNFR1, inducible NO synthase, and ICAM-1 were up-regulated in brains of lupus mice. Crry prevented the increased expression of these inflammatory molecules, indicating that the changes were complement dependent. Furthermore, microarray analysis revealed complement-dependent up-regulation of glutamate receptor (AMPA GluR) expression in lupus brains, which was also validated for AMPA-GluR1 mRNA and protein. Our results clearly demonstrate that apoptosis is a prominent feature in lupus brains. Complement activation products either directly and/or indirectly through TNFR1, ICAM-1, inducible NO synthase, and AMPA-GluR, all of which were altered in MRL/lpr mouse brains, have the potential to induce such apoptosis. These findings present the exciting possibility that complement inhibition is a therapeutic option for lupus cerebritis. PMID- 16339573 TI - Topical superantigen exposure induces epidermal accumulation of CD8+ T cells, a mixed Th1/Th2-type dermatitis and vigorous production of IgE antibodies in the murine model of atopic dermatitis. AB - Patients with atopic dermatitis (AD) have repeated cutaneous exposure to both environmental allergens and superantigen-producing strains of Staphylococcus aureus. We used a murine model of AD to investigate the role of staphylococcal enterotoxin B (SEB) in the modulation of allergen-induced skin inflammation. Mice were topically exposed to SEB, OVA, a combination of OVA and SEB (OVA/SEB), or PBS. Topical SEB and OVA/SEB exposure induced epidermal accumulation of CD8+ T cells and TCRVbeta8+ cells in contrast to OVA application, which induced a mainly dermal infiltration of CD4+ cells. SEB and OVA/SEB exposure elicited a mixed Th1/Th2-associated cytokine and chemokine expression profile within the skin. Restimulation of lymph node cells from OVA- and OVA/SEB-exposed mice with OVA elicited strong production of IL-13 protein, whereas substantial amounts of IFN gamma protein were detected after SEB stimulation of cells derived from SEB- or OVA/SEB-exposed mice. Topical SEB treatment elicited vigorous production of SEB specific IgE and IgG2a Abs and significantly increased the production of OVA specific IgE and IgG2a Abs. The present study shows that topical exposure to SEB provokes epidermal accumulation of CD8+ T cells, a mixed Th2/Th1 type dermatitis and vigorous production of specific IgE and IgG2a Abs, which can be related to the chronic phase of atopic skin inflammation. PMID- 16339574 TI - Aberrant expression of the autoantigen heterogeneous nuclear ribonucleoprotein-A2 (RA33) and spontaneous formation of rheumatoid arthritis-associated anti-RA33 autoantibodies in TNF-alpha transgenic mice. AB - Human TNF-alpha transgenic (hTNFtg) mice develop erosive arthritis closely resembling rheumatoid arthritis (RA). To investigate mechanisms leading to pathological autoimmune reactions in RA, we examined hTNFtg animals for the presence of RA-associated autoantibodies including Abs to citrullinated epitopes (anti-cyclic citrullinated peptide), heterogeneous nuclear ribonucleoprotein (hnRNP)-A2 (anti-RA33), and heat shock proteins (hsp) (anti-hsp). Although IgM anti-hsp Abs were detected in 40% of hTNFtg and control mice, IgG anti-hsp Abs were rarely seen, and anti-cyclic citrullinated peptide Abs were not seen at all. In contrast, >50% of hTNFtg mice showed IgG anti-RA33 autoantibodies, which became detectable shortly after the onset of arthritis. These Abs were predominantly directed to a short epitope, which was identical with an epitope previously described in MRL/lpr mice. Incidence of anti-RA33 was significantly decreased in mice treated with the osteoclast inhibitor osteoprotegerin and also in c-fos-deficient mice lacking osteoclasts. Pronounced expression of hnRNP-A2 and a smaller splice variant was seen in joints of hTNFtg mice, whereas expression was low in control animals. Although the closely related hnRNP-A1 was also overexpressed, autoantibodies to this protein were infrequently detected. Because expression of hnRNP-A2 in thymus, spleen, brain, and lung was similar in hTNFtg and control mice, aberrant expression appeared to be restricted to the inflamed joint. Finally, immunization of hTNFtg mice with recombinant hnRNP-A2 or a peptide harboring the major B cell epitope aggravated arthritis. These findings suggest that overproduction of TNF-alpha leads to aberrant expression of hnRNP-A2 in the rheumatoid joint and subsequently to autoimmune reactions, which may enhance the inflammatory and destructive process. PMID- 16339575 TI - Mcl-1 is essential for the survival of synovial fibroblasts in rheumatoid arthritis. AB - Mcl-1 is a Bcl-2-family, antiapoptotic molecule that is critical for the survival of T and B lymphocytes and macrophages; however, its role in nonhemopoietic cells remains to be fully elucidated. The current study focuses on the role of Mcl-1 in rheumatoid arthritis (RA). Mcl-1 was strongly expressed in the synovial lining and was increased in the sublining fibroblasts of patients with RA, compared with control synovial tissue. The expression of Mcl-1 in sublining fibroblasts correlated with the degree of inflammation and TNF-alpha, and IL-1beta treatment of cultured synovial fibroblasts resulted in the increased expression of Mcl-1 at the mRNA and protein levels. Mcl-1 was critical for the survival of RA synovial fibroblasts, because the forced reduction of Mcl-1 using a Mcl-1 antisense expressing adenoviral vector induced apoptotic cell death, which was mediated through Bax, Bak, and Bim. These observations document a critical role for Mcl-1 in protecting against apoptosis in RA and suggest that Mc1-1 is a potential therapeutic target in this disease. PMID- 16339576 TI - Saliva from Lutzomyia longipalpis induces CC chemokine ligand 2/monocyte chemoattractant protein-1 expression and macrophage recruitment. AB - Saliva of bloodfeeding arthropods has been incriminated in facilitating the establishment of parasite in their host. We report on the leukocyte chemoattractive effect of salivary gland homogenate (SGH) from Lutzomyia longipalpis on saliva-induced inflammation in an air pouch model. SGH (0.5 pair/animal) was inoculated in the air pouch formed in the back of BALB/c or C57BL/6 mice. L. longipalpis SGH induced a significant influx of macrophages in BALB/c but not in C57BL/6 mice. SGH-induced cell recruitment reached a peak at 12 h after inoculation and was higher than that induced by the LPS control. This differential cell recruitment in BALB/c mice was directly correlated to an increase in CCL2/MCP-1 expression in the air pouch lining tissue. In fact, treatment with bindarit, an inhibitor of CCL2/MCP-1 synthesis, and also with a specific anti-MCP-1 mAb resulted in drastic reduction of macrophage recruitment and inhibition of CCL2/MCP-1 expression in the lining tissue. CCL2/MCP-1 production was also seen in vitro when J774 murine macrophages were exposed to L. longipalpis SGH. The SGH effect was abrogated by preincubation with serum containing anti-SGH IgG Abs as well as in mice previously sensitized with L. longipalpis bites. Interestingly, the combination of SGH with Leishmania chagasi induced an increased recruitment of neutrophils and macrophages when compared with L. chagasi alone. Taken together these results suggest that SGH not only induces the recruitment of a greater number of macrophages by enhancing CCL2/MCP 1 production but also synergizes with L. chagasi to recruit more inflammatory cells to the site of inoculation. PMID- 16339577 TI - Reduced allergenic potency of VR9-1, a mutant of the major shrimp allergen Pen a 1 (tropomyosin). AB - The major shrimp allergen, tropomyosin, is an excellent model allergen for studying the influence of mutations within the primary structure on the allergenic potency of an allergen; Pen a 1 allows systematic evaluation and comparison of Ab-binding epitopes, because amino acid sequences of both allergenic and nonallergenic tropomyosins are known. Individually recognized IgE Ab-binding epitopes, amino acid positions, and substitutions critical for IgE Ab binding were identified by combinatorial substitution analysis, and 12 positions deemed critical were mutated in the eight major epitopes. The mutant VR9-1 was characterized with regard to allergenic potency by mediator release assays using sera from shrimp-allergic subjects and sera from BALB/c, C57BL/6J, C3H/HeJ, and CBA/J mice sensitized with shrimp extract using alum, cholera toxin, and Bordetella pertussis, as adjuvants. The secondary structure of VR9-1 was not altered; however, the allergenic potency was reduced by 90-98% measuring allergen specific mediator release from humanized rat basophilic leukemia (RBL) cells, RBL 30/25. Reduced mediator release of RBL-2H3 cells sensitized with sera from mice that were immunized with shrimp extract indicated that mice produced IgE Abs to Pen a 1 and to the same epitopes as humans did. In conclusion, data obtained by mapping sequential epitopes were used to generate a Pen a 1 mutant with significantly reduced allergenic potency. Epitopes that are relevant for human IgE Ab binding are also major binding sites for murine IgE Abs. These results indicate that the murine model might be used to optimize the Pen a 1 mutant for future therapeutic use. PMID- 16339579 TI - Differential expression of alternatively spliced transcripts of HLA-G in human preimplantation embryos and inner cell masses. AB - It has been reported that preimplantation human embryos secrete HLA-G, and the levels may be predictive of their ability to implant. However, it is not known which of the membrane-bound (HLA-G 1-4) and soluble (HLA-G 5-6) alternatively spliced forms are present, nor the developmental stage at which they appear. Therefore, we have investigated HLA-G mRNA isoform expression on single embryos at the two-, four-, six-, and eight-cell, morula, and blastocyst stages. The percentage of embryos expressing each HLA-G isoform mRNA increased with developmental stage, but contrary to expectation, HLA-G5 mRNA was not detected in single two- to eight-cell embryos and was only expressed by 20% of morulae and blastocysts. Similarly, soluble HLA-G6 mRNA was not detected until the blastocyst stage and then in only one-third of embryos. In contrast, labeling with MEM G/9 Ab (specific for HLA-G1 and -G5) was observed in 15 of 20 two- to eight-cell embryos and 5 of 5 blastocysts. This disparity between mRNA and protein may be due to HLA-G protein remaining from maternal oocyte stores produced before embryonic genome activation and brings into question the measurement of soluble HLA-G for clinical evaluation of embryo quality. Although HLA-G is expressed in the preimplantation embryo, later it is primarily expressed in the invasive trophoblast of the placenta rather than the fetus. Therefore, we have investigated whether down-regulation of HLA-G first occurs in the inner cell mass (precursor fetal cells) of the blastocyst and, in support of this concept, have shown the absence HLA-G1 and -G5 protein and mRNA. PMID- 16339578 TI - CD8+ T cell-mediated HLA-A*0201-restricted cytotoxicity to transaldolase peptide 168-176 in patients with multiple sclerosis. AB - Transaldolase (TAL) is expressed at selectively high levels in oligodendrocytes and targeted by autoreactive T cells of patients with multiple sclerosis (MS). Among 14 TAL peptides with predicted HLA-A2 binding, TAL 168-176 (LLFSFAQAV, TALpep) exhibited high affinity for HLA-A2. Prevalence of HLA-A2-restricted CD8+ T cells specific for TALpep was increased in PBMC of HLA-A2+ MS patients, as compared with HLA-A2- MS patients, HLA-A2+ other neurological disease patients, and HLA-A2+ healthy donors. HLA-A*0201/TALpep tetramers detected increased frequency of TAL-specific CD8+ T cells, and precursor frequency of TAL-specific IFN-gamma-producing T cells was increased in each of seven HLA-A2+ MS patients tested. Stimulation by TALpep or rTAL of PBMC from HLA-A2+ MS patients elicited killing of TALpep-pulsed HLA-A2-transfected HmyA2.1 lymphoma cells, but not HLA A3-transfected control HmyA3.1 targets. Without peptide pulsing of targets, HLA A2-transfected, but not control MO3.13 oligodendroglial cells, expressing high levels of endogenous TAL, were also killed by CD8+ CTL of MS patients, indicating recognition of endogenously processed TAL. TCR Vbeta repertoire analysis revealed use of the TCR Vbeta14 gene by T cell lines (TCL) of MS patients generated via stimulation by TAL- or TALpep-pulsed APCs. All TAL-specific TCL-binding HLA A*0201/TALpep tetramers expressed TCR Vbeta14 on the cell surface. Moreover, Ab to TCR Vbeta14 abrogated cytotoxicity by HLA-A2-restricted TAL-specific TCL. Therefore, TAL-specific CTL may serve as a novel target for therapeutic intervention in patients with MS. PMID- 16339580 TI - Cloning the arterial IgA antibody response during acute Kawasaki disease. AB - Kawasaki disease (KD) is the most common acquired cardiac disease in children in developed nations. The etiology of KD is unknown but likely to be a ubiquitous microbial agent. Previously, we showed that oligoclonal IgA plasma cells infiltrate coronary arteries and other inflamed tissues in acute KD. We demonstrated that a synthetic Ab made using an alpha H chain sequence prevalent in acute KD arterial tissue detected Ag in acute KD coronary arteries, lung, and other inflamed tissues and that Ag localized to cytoplasmic inclusion bodies in the acute KD ciliated bronchial epithelium. In this study, we synthesized a panel of mAbs from alpha and kappa chain sequences present in the KD arterial wall and tested the Abs for binding to acute KD tissues. We report that all of the synthetic mAbs that bind to acute KD tissues detect Ag in cytoplasmic inclusion bodies in the acute KD ciliated bronchial epithelium. Abs made from alpha sequences that were prevalent in KD arterial tissue show stronger binding to acute KD tissues than Abs made from less prevalent sequences. These findings highlight the likely importance of the inclusion bodies in the etiopathogenesis of acute KD, confirm that the IgA Ab response in acute KD is Ag driven, and demonstrate the usefulness of cloning the Ab response in diseased tissues to identify disease-relevant Ags. PMID- 16339581 TI - Global natural regulatory T cell depletion in active systemic lupus erythematosus. AB - The immune defect that could account for the multisystemic involvement that characterizes systemic lupus erythematosus (SLE) remains unknown. We hypothesized that iterative disease flares correspond to a recurrent defect in the peripheral immune suppression exerted by naturally occurring T regulatory cells (Tregs). Surprisingly, Tregs isolated from lupus patients show the same phenotypic and functional characteristics as corresponding cells found in healthy controls. A decrease in the proportion of circulating Tregs among other CD4+ T cells is nevertheless evidenced in active patients when this group is compared with healthy controls (0.57 +/- 0.24%, n = 45 vs 1.29 +/- 0.38%, n = 82, p < 0.0001) or with inactive patients (1.22 +/- 0.67%, n = 62, p < 0.0001). In contrast, the proportion of Tregs in other systemic autoimmune diseases such as primary Sjogren syndrome and inflammatory myopathy does not significantly differ from controls' values (1.15 +/- 0.46%, n = 21, p = 0.09 and 1.16 +/- 0.44%, n = 16, p = 0.43, respectively). Lupus Tregs do not accumulate in either the lymph nodes or the diseased kidneys and are not killed by a circulating soluble factor, but demonstrate in vitro a heightened sensitivity to Fas-induced apoptosis. Finally, we show that the extent of Treg depletion correlates with the clinical severity of the flare. SLE flares are therefore associated with a global Treg depletion and not with a phenomenon of tissue redistribution. In summary, we suggest that the physiopathology of SLE could be tied to a defect in the homeostatic control of the Treg subpopulation. PMID- 16339582 TI - CXCL10 DNA vaccination prevents spontaneous diabetes through enhanced beta cell proliferation in NOD mice. AB - CXCL10, a chemokine for Th1 cells, is involved in the pathogenesis of various Th1 dominant autoimmune diseases. Type 1 diabetes is considered to be a Th1-dominant autoimmune disease, and a suppressive effect of CXCL10 neutralization on diabetes development has been reported in a cyclophosphamide-induced accelerated diabetes model through induction of beta cell proliferation. However, intervention in a diabetes model might bring about opposite effects, depending on the timing, amount, or method of treatment. In the present study, we examined the effect of CXCL10 neutralization in a "spontaneous diabetes" model of NOD mice, using CXCL10 DNA vaccination (pCAGGS-CXCL10). pCAGGS-CXCL10 treatment in young NOD mice induced the production of anti-CXCL10 Ab in vivo and suppressed the incidence of spontaneous diabetes, although this treatment did not inhibit insulitis or alter the immunological response. pCAGGS-CXCL10 treatment enhanced the proliferation of pancreatic beta cells, resulting in an increase of beta cell mass in this spontaneous diabetes model as well. Therefore, CXCL10 neutralization is suggested to be useful for maintaining beta cell mass at any stage of autoimmune diabetes. PMID- 16339583 TI - Immunization with short peptides from the 60-kDa Ro antigen recapitulates the serological and pathological findings as well as the salivary gland dysfunction of Sjogren's syndrome. AB - Sjogren's syndrome is a poorly understood autoimmune inflammatory illness that affects the salivary and lacrimal glands as well as other organ systems. We undertook the present study to determine whether mice immunized with short peptides from the 60-kDa Ro (or SSA) Ag, which is a common target of the autoimmunity of Sjogren's syndrome, develop an illness similar to Sjogren's syndrome. BALB/c mice were immunized with one of two short peptides from 60-kDa Ro that are know to induce epitope spreading. The animals were analyzed for the presence of anti-Ro and anti-La (or SSB) in the sera by immunoblot and ELISA. Salivary glands were collected and examined by histology after H&E staining. Salivary lymphocytes were purified and studied for cell surface makers by fluorescence-activated cell sorting. Timed stimulated salivary flow was measured. As reported previously, BALB/c mice immunized with 60-kDa Ro peptides developed an immune response directed against the entire Ro/La ribonucleoprotein particle that was similar to that found in humans with lupus or Sjogren's syndrome. Functional studies showed a statistical decrease in salivary flow in immunized mice compared with controls. Furthermore, there were lymphocytic infiltrates in the salivary glands of immunized animals that were not present in controls. The infiltrates consisted of both CD4- and CD8+ T lymphocytes as well as B lymphocytes. BALB/c mice immunized with 60-kDa Ro peptides develop anti-Ro, salivary gland lymphocyte infiltrates, and salivary dysfunction that is highly reminiscent of human Sjogren's syndrome. PMID- 16339584 TI - Functional and phenotypic characterization of CD57+CD4+ T cells and their association with HIV-1-induced T cell dysfunction. AB - HIV-1 replication is associated with reduced or absent HIV-1-specific CD4+ T cell proliferation and skewing of HIV-1-specific CD4+ T cells toward an IFN-gamma producing, CCR7- phenotype. The CCR7- T cell population is heterogeneous and can be subdivided based on the expression of CD57. Although CD57 expression on CD8+ T cells is associated with proliferation incompetence and replicative senescence, less is known about the function of CD57-expressing CD4+ T cells. In this study, the frequency, phenotype, and function of CD57+CD4+ T cells were evaluated in 25 HIV-1-infected subjects and 10 seronegative controls. CD57+CD4+ T cells were found to be proliferation incompetent, even after strong mitogen stimulation. Percentages of CD4+ T cells that expressed CD57 were significantly higher in untreated HIV-1-infected subjects than in HIV-1-seronegative donors, and CD57 expression did not normalize in subjects receiving at least 6 mo of effective antiretroviral therapy. CD57 was predominately expressed on the CCR7- fraction of the CD4+ T cell compartment and accounted for the majority of cells in the CCR7 CD45RA+ population from untreated HIV-1-infected subjects. HIV-1-specific CD4+ T cells producing only IFN-gamma had the highest expression of CD57, whereas few cells producing IL-2 alone expressed CD57. These findings further define a novel population of proliferation-incompetent CD4+ T cells that are generated in the presence of chronic Ag exposure. A better understanding of the generation and persistence of CD57+ T cells in HIV-1 infection could provide important insights into the immunopathogenesis of this disease. PMID- 16339585 TI - Expression and activity of NOX5 in the circulating malignant B cells of hairy cell leukemia. AB - Hairy cells (HCs) are mature malignant B cells that contain a number of constitutively active signaling molecules including GTP-bound Rac1, protein kinase C, and Src family kinases. Because Rac1 is a component of the reactive oxidant species (ROS)-generating NADPH oxidase system, we investigated the role of this GTPase in ROS production in HCs. In this study, we show that ROS production in HCs involves a flavin-containing oxidase dependent on Ca2+, but not on GTP-Rac1 or protein kinase C. This suggests the involvement of the nonphagocytic NADPH oxidase NOX5, an enzyme found in lymphoid tissues, but not in circulating lymphocytes. By using RT-PCR and Southern and Western blotting and by measuring superoxide anion production in membrane fractions in the absence of cytosolic components, we demonstrate for the first time that HCs (but not circulating normal B cells or some other lymphoid cell types) express NOX5. We also demonstrate that inhibition of NADPH oxidase in HCs results in a selective increase in the activity of Src homology region 2 domain-containing phosphatase 1 (SHP-1). Furthermore, SHP-1 in HCs coimmunoprecipitates with tyrosine phosphorylated CD22 and localizes in the same cellular compartment as NOX5. This allows the inactivation of SHP-1 by NOX5-generated ROS and contributes to the maintenance of the constitutive activation of HCs. PMID- 16339586 TI - Immunodominance of poxviral-specific CTL in a human trial of recombinant-modified vaccinia Ankara. AB - Many recombinant poxviral vaccines are currently in clinical trials for cancer and infectious diseases. However, these agents have failed to generate T cell responses specific for recombinant gene products at levels comparable with T cell responses associated with natural viral infections. The recent identification of vaccinia-encoded CTL epitopes, including a new epitope described in this study, allows the simultaneous comparison of CTL responses specific for poxviral and recombinant epitopes. We performed detailed kinetic analyses of CTL responses in HLA-A*0201 patients receiving repeated injections of recombinant modified vaccinia Ankara encoding a string of melanoma tumor Ag epitopes. The vaccine driven CTL hierarchy was dominated by modified vaccinia Ankara epitope-specific responses, even in patients who had not received previous smallpox vaccination. The only recombinant epitope that was able to impact on the CTL hierarchy was the melan-A26-35 analog epitope, whereas responses specific for the weaker affinity epitope NY-ESO-1(157-165) failed to be expanded above the level detected in prevaccination samples. Our results demonstrate that immunodominant vaccinia specific CTL responses limit the effectiveness of poxviruses in recombinant vaccination strategies and that more powerful priming strategies are required to overcome immunodominance of poxvirus-specific T cell responses. PMID- 16339588 TI - Commentary: prescient visions of public health from Cornaro to Breslow. PMID- 16339590 TI - Public health aspects of weight control.Commentary: What's past is prologue. PMID- 16339591 TI - Commentary: the importance of addressing the rise of overweight and obesity- progress or lack of action during the last fifty years? PMID- 16339592 TI - The epidemiology of overweight and obesity: Commentary: counterpoint to Campos et al. PMID- 16339593 TI - Commentary: the epidemic of obesity--what's in a name? PMID- 16339594 TI - Commentary: current perspectives on obesity and health: black and white, or shades of grey? PMID- 16339595 TI - Commentary: obesity claims and controversies. PMID- 16339596 TI - Commentary: obesity--public health crisis, moral panic or a human rights issue? PMID- 16339597 TI - Commentary: there is a public health crisis--its not fat on the body but fat in the mind and the fat of profits. PMID- 16339598 TI - Commentary: understanding the epidemiology of overweight and obesity--a real global public health concern. PMID- 16339599 TI - The epidemiology of overweight and obesity: public health crisis or moral panic? PMID- 16339600 TI - What aspects of body fat are particularly hazardous and how do we measure them? PMID- 16339601 TI - Commentary: Obesity and mortality--light at the end but still a long tunnel. PMID- 16339602 TI - Commentary: obesity is not a newly recognized public health problem--a commentary of Breslow's 1952 paper on 'public health aspects of weight control'. PMID- 16339604 TI - Public health aspects of weight control. 1952. PMID- 16339605 TI - Commentary: 'culture', cultural explanations and causality. PMID- 16339606 TI - An open-label randomized trial comparing itraconazole oral solution with fluconazole oral solution for primary prophylaxis of fungal infections in patients with haematological malignancy and profound neutropenia. AB - OBJECTIVES: This trial studied the efficacy and safety of itraconazole and fluconazole in the prevention of invasive fungal infections in neutropenic patients with haematological malignancies. PATIENTS AND METHODS: An 8 week, open label, randomized, parallel-group, multicentre trial comparing itraconazole oral solution (2.5 mg/kg twice daily; N=248) with fluconazole oral solution or capsules (400 mg daily; N=246) in 494 patients with anticipated profound neutropenia (i.e. neutrophil count expected to be <500 cells/mm3 for at least 10 days) from tertiary care centres. RESULTS: Invasive fungal infections were reported for 4 out of 248 patients (1.6%) in the itraconazole group and 5 out of 246 patients (2.0%) in the fluconazole group. Invasive Aspergillus infections were proven for 2 out of 248 patients (0.8%) in the itraconazole group and 3 out of 246 patients (1.2%) in the fluconazole group. For both the ITT and profoundly neutropenic populations, no differences were detected between treatment groups in proven or suspected invasive fungal infections or other endpoints. The mortality rates owing to proven invasive fungal infections were 2 out of 248 patients (0.8%) for the itraconazole group and 3 out of 246 patients (1.2%) for the fluconazole group. There was also no difference between treatment groups in the number of patients who recovered from neutropenia or in the duration of neutropenia. More discontinuation of drug intake owing to nausea and more hypokalaemia occurred in the itraconazole group, other adverse events and the total number of adverse events were similar in both groups. CONCLUSIONS: In this study there were no differences in the efficacy and safety of itraconazole and fluconazole prophylaxis in neutropenic patients with haematological malignancies. PMID- 16339607 TI - Macrolide inactivation gene cluster mphA-mrx-mphR adjacent to a class 1 integron in Aeromonas hydrophila isolated from a diarrhoeic pig in Oklahoma. AB - OBJECTIVES: To characterize a multidrug-resistant Aeromonas hydrophila isolate (CVM861) that possesses a high-level macrolide inactivation gene cluster (mphA mrx-mphR), previously only reported in Escherichia coli. METHODS: PCR fragment length mapping, gene sequencing and Southern blotting were used to map the mphA mrx-mphR gene cluster and flanking elements in CVM861. Conjugation experiments were done to determine whether the multidrug resistance genetic element was mobile. RESULTS: The mphA-mrx-mphR gene cluster mapped downstream of a class 1 integron and upstream of an aph(3') gene, and was present on a Tn21-like element. The gene order determined by sequencing was intI1-dhfrXII-orfF-aadA2-qacDeltaE sul1-orf5Delta178-tnpA-mphR-mrx-mphA. Horizontal transmission of high-level macrolide resistance from CVM861 to E. coli 47011 was inconsistent; however, a composite plasmid possessing the mphA gene cluster was transferred at a conjugation frequency of 2.02 x 10(-5) per recipient. CONCLUSIONS: An mphA-mrx mphR gene cluster was present downstream of the In2 integron located on a Tn21 like transposon in an A. hydrophila isolate. Whether this recombination event resulted in the truncation of the orf5 sequence is unknown. The presence of other resistance genes downstream of the mphA-mrx-mphR gene cluster suggests that multiple recombination events have occurred on this genetic element. This is the first known report of the mphA-mrx-mphR gene cluster carried by A. hydrophila and the first known isolation of this cluster in the United States. PMID- 16339608 TI - Should the CATIE study be a wake-up call? PMID- 16339610 TI - Datapoints: trends in ADHD and stimulant use among adults: 1995-2002. PMID- 16339611 TI - Personal accounts: my experiences as a psychiatric patient in the 1960s. PMID- 16339612 TI - Disparities in depression treatment for Latinos and site of care. AB - OBJECTIVE: This study examined the impact of patient characteristics and source of care on differences between whites and Latinos in use and quality of depression treatment in managed primary care settings. METHODS: Data were examined for 1,175 patients (398 Latinos and 777 whites) in 46 managed primary care practices who screened positive for probable depressive disorder. Patient baseline assessments were used to compile sociodemographic and clinical characteristics and to derive variables for receipt of any depression care and depression care that met minimum guidelines (antidepressant use or specialty counseling) in the past six months. Clinics were classified by the percentage of their patient population that consisted of Latinos to determine whether patients in highly Latino clinics reported lower rates of care. Predictors of use and quality of depression care were examined by using logistic regression. RESULTS: Rates of receipt of any depression care and guideline-level depression care were low, and Latinos were less than half as likely as whites to receive such care, even after the analyses controlled for independent predictors (that is, younger age, higher educational level, current unemployment, more comorbid medical illness, and a diagnosis of a depressive or anxiety disorder). The likelihood of receiving any care or care that met guidelines did not significantly vary according to whether clinics served a low, moderate, or high percentage of Latinos. CONCLUSIONS: Disparities in depression care for Latinos were not attributable to sociodemographic and clinical characteristics, and they were not attributable to receiving care in clinics that served ethnically similar or dissimilar clientele. These findings suggest that other patient or provider factors may be responsible. PMID- 16339613 TI - Predictors of adequacy of depression management in the primary care setting. AB - OBJECTIVES: Depression is most commonly treated in the primary care setting. However, most studies have shown that depression care in this setting is often inadequate. This study examined the adequacy of antidepressant treatment and overall depression management by primary care physicians and identified patient characteristics related to inadequate care. METHODS: Adequacy of depression care among patients with depression who presented to a primary care office was evaluated by using physicians' self-reports and medication prescription data. Adequacy of depression care was measured in two ways: adequacy of the current medication trial was measured by using the Antidepressant Treatment and History Form (ATHF), and adequacy of overall management by the physician was measured by using an algorithm developed for this study. The association was examined between patient characteristics and adequacy of the medication trial or depression management. RESULTS: Data were gathered for 389 patients with depression. Overall, 71 percent of patients had adequate ATHF scores, and 75 percent were judged to receive adequate depression management. No significant differences in adequacy were seen on the basis of race, age, or gender. When depressive symptoms were almost absent or extremely severe, 91 percent of patients were adequately managed; in contrast, when symptoms were mild, mild to moderate, or moderate 69 percent of patients were adequately managed. Additionally, specific medications, such as sertraline and fluoxetine, were associated with a lower likelihood of an adequate ATHF score. CONCLUSIONS: A majority of these primary care patients were adequately treated for depression, with no detectable disparity related to race or age. However, mild to moderate depressive symptoms (as opposed to remitted or severe symptoms) and specific medications were associated with a lower rate of adequacy. These findings have implications for ways that primary care physicians could be trained to more adequately manage the spectrum of severity of depression. PMID- 16339614 TI - Use of health care services among persons who screen positive for bipolar disorder. AB - OBJECTIVE: This study examined patterns of diagnosis, consultation, and treatment of persons who screened positive for bipolar disorder. METHODS: An impact survey was mailed to a representative subset of 3,059 individuals from a large U.S. population-based study that utilized the Mood Disorder Questionnaire (MDQ). RESULTS: Respondents who screened positive on the MDQ (reported the presence of seven of 13 symptoms of bipolar disorder, the co-occurrence of at least two symptoms, and moderate or severe symptom-related impairment) (N=1,167) had consulted a health care provider more often in the previous year than those who screened negative (reported six or fewer symptoms regardless of symptom co occurrence or impairment) (N=1,283). Psychiatrists and primary care physicians failed to detect or misdiagnosed bipolar disorder among 53 percent and 78 percent of patients, respectively, who screened positive for bipolar disorder. The most commonly used psychotropic medications during the previous 12 months among those who screened positive were antidepressants alone (32 percent), followed by lithium and anticonvulsant mood stabilizers (20 percent), antidepressants in combination with other psychotropics (19 percent), hypnotics (19 percent), and antipsychotics (9 percent). In the preceding 12 months, respondents who screened positive on the MDQ had greater use of psychiatric hospitals, emergency departments, and urgent care centers and also had more outpatient visits to primary care physicians, psychiatrists, and alcohol treatment centers than those who screened negative. CONCLUSIONS: The results of this study suggest that bipolar disorder is an underdiagnosed and often inappropriately treated illness associated with significant use of health care resources. PMID- 16339615 TI - Suicidal ideation and pharmacotherapy among STEP-BD patients. AB - OBJECTIVE: Little is known about the effects of lithium on suicidal ideation or about the possible antisuicidal effects of divalproex, second-generation antipsychotics, or antidepressants among persons with bipolar disorder. METHODS: Using a cross-sectional design, the authors examined patterns of psychotropic drug use relative to suicidal ideation among 1,000 patients with bipolar disorder in the National Institute of Mental Health's Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD). RESULTS: The presence of suicidal ideation was similar between patients who were taking any lithium and those who were not (22.2 percent and 25.8 percent, respectively) and between those who were taking any divalproex and those who were not (20.3 percent and 21.5 percent). Suicidal ideation was significantly more prevalent among patients who were taking a second-generation antipsychotic than those who were not (26 percent and 17 percent) and those who were taking an antidepressant and those who were not (25 percent and 14 percent). After other variables had been controlled for, lithium prescriptions were significantly more common among patients who had suicidal ideation. CONCLUSIONS: Among patients with bipolar disorder who have suicidal ideation, antidepressants and second-generation antipsychotics appear to be prescribed by community practitioners more often than other medications, with lithium reserved for those with more severe illness characteristics. PMID- 16339616 TI - Baseline predictors of serious adverse events at one year among patients with bipolar disorder in STEP-BD. AB - OBJECTIVE: Little is known about serious adverse events that occur in the context of clinical care for bipolar disorder. This study examined predictors of serious adverse events in a large cohort of patients with bipolar disorder. METHODS: Types and frequency of serious adverse events were tabulated on the basis of data from the first 1,000 participants in the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD). STEP-BD follows a large cohort of patients longitudinally. Treatments are provided as deemed most appropriate by the treating psychiatrist. Logistic regression was used to identify variables present at study entry that were associated with the occurrence of a serious adverse event within one year. RESULTS: A total of 161 serious adverse events were reported among 118 participants. The most frequent serious adverse event was hospitalization for suicidal ideation (44 participants, or 27 percent). A greater number of previous psychiatric medications was predictive of the occurrence of a psychiatric serious adverse event among participants who had less than one year of study follow-up data. A history of psychosis, current substance abuse, lower household income, and a fully syndromic baseline mood state were associated with the occurrence of a psychiatric serious adverse event among participants who had one full year of follow-up data. CONCLUSIONS: These data identify clinical and demographic factors that warrant closer clinical follow-up, because these factors may be predictive of poor outcomes in the following year, even in the context of expert care. PMID- 16339618 TI - Racial differences in syndromal and subsyndromal depression in an older urban population. AB - OBJECTIVE: The authors explored racial differences in the prevalence of depression and its associated factors among older persons. METHODS: Using 1990 census data for Brooklyn, New York, the authors attempted to interview all cognitively intact persons age 55 years and older in randomly selected block groups. The sample was weighted by ethnicity and gender. The authors adapted George's Social Antecedent Model of Depression to allow examination of 20 independent variables and the nominal dependent variable consisting of three levels of depression. The data were analyzed with SUDAAN. RESULTS: Syndromal depression was found among 8 percent of blacks and 10 percent of whites. Subsyndromal depression was found among 13 percent of blacks and 28 percent of whites. No racial differences were found in rates of syndromal depression, but significant racial differences were found in rates of subsyndromal depression and of any type of depression. Nonlinear effects on both types of depression were found, and higher levels of stress had a greater impact on whites than on blacks. The racial difference in subsyndromal depression was explained by its lower prevalence among French-speaking African Caribbeans. Many racial differences were found in the variables associated with syndromal and subsyndromal depression. CONCLUSIONS: Race had an independent effect on the rate of subsyndromal depression and an interactive effect with stress on the rate of both syndromal and subsyndromal depression. For each racial group, different elements may play a role in the etiology, maintenance, and relief of depression. The findings underscore the importance of recognizing within-group and between-group racial differences in depression. PMID- 16339617 TI - Racial differences in the treatment of veterans with bipolar disorder. AB - OBJECTIVES: The authors examined whether African Americans, compared with whites, received guideline-concordant care for bipolar I disorder. METHODS: A retrospective analysis was conducted of data for patients who received a diagnosis of bipolar I disorder in fiscal year 2001 and received care in facilities in the Department of Veterans Affairs (VA) mid-Atlantic region. Indicators of guideline-concordant care were based on prescription data and data on utilization of inpatient and outpatient services from VA databases. RESULTS: A total of 2,316 patients with a diagnosis of bipolar I disorder were identified. Their mean age was 52 years; 9.4 percent (N=218) were women, and 13.1 percent (N=303) were African American. Overall, mood stabilizers were prescribed for 74.6 percent (N=1,728) of the patients; 67.1 percent (N=1,554) had an outpatient mental health visit within 90 days after the index diagnosis, and 54.3 percent (N=1,258) had an outpatient visit within 30 days after discharge from a psychiatric hospitalization. Multivariate logistic regression analyses with adjustment for sociodemographic and facility factors revealed that African Americans were less likely than whites to have an outpatient follow-up visit within 90 days after the index diagnosis. Race was not associated with receipt of mood stabilizers or use of outpatient services after a hospital discharge. CONCLUSIONS: Although a majority of patients received guideline-concordant care for bipolar disorder, potential gaps in continuity of outpatient care may exist for African-American patients. PMID- 16339619 TI - The Cornell Service Index as a measure of health service use. AB - OBJECTIVE: This article describes the development, administration, and reliability of the Cornell Services Index (CSI), a new instrument that measures health service use. The CSI was developed to create a standardized measure of the quantity and characteristics (for example, site and provider) of services used by adults. Descriptive data are provided to illustrate the application of the CSI in a community sample of adults who were newly admitted to outpatient mental health clinics. These data provide information about the pathways to care. METHODS: The interrater and test-retest reliability of the CSI were evaluated by using a sample of 40 adults who were seeking mental health treatment. Descriptive data on service use in a sample of 1,279 adults seeking care in outpatient mental health clinics was provided to demonstrate the application of the CSI. RESULTS: The CSI is a portable, easy to use, and brief assessment of service use. It has good interrater and test-retest reliability among adults without cognitive impairment. In the three months before seeking care, 31 percent of the adults interviewed had made a mental health visit, 36 percent had been hospitalized, and more than half (59 percent) had made a medical visit. Twenty-three percent of adults had sought care from a hospital's emergency department. CONCLUSIONS: The CSI is a reliable method to assess health service use for adults. The measure can extend assessment of use beyond the traditional mental health service use questions and provide a snapshot of service use patterns across types, providers, and sites of service among adults who seek mental health care. PMID- 16339620 TI - Performance measures for early psychosis treatment services. AB - OBJECTIVE: This study examined the feasibility of identifying performance measures for early psychosis treatment services and obtaining consensus for these measures. The requirements of the study were that the processes used to identify measures and gain consensus should be comprehensive, be reproducible, and reflect the perspective of multiple stakeholders in Canada. METHODS: The study was conducted in two stages. First a literature review was performed to gather articles published from 1995 to July 2002, and experts were consulted to determine performance measures. Second, a consensus-building technique, the Delphi process, was used with nominated participants from seven groups of stakeholders. Twenty stakeholders participated in three rounds of questionnaires. The degree of consensus achieved by the Delphi process was assessed by calculating the semi-interquartile range for each measure. RESULTS: Seventy-three performance measures were identified from the literature review and consultation with experts. The Delphi method reduced the list to 24 measures rated as essential. This approach proved to be both feasible and cost-effective. CONCLUSIONS: Despite the diversity in the backgrounds of the stakeholder groups, the Delphi technique was effective in moving participants' ratings toward consensus through successive questionnaire rounds. The resulting measures reflected the interests of all stakeholders. PMID- 16339621 TI - Commentary: selecting performance measures by consensus: an appropriate extension of the delphi method? PMID- 16339622 TI - The clinical strategies implementation scale to measure implementation of treatment in mental health services. AB - OBJECTIVE: The authors describe the development of the Clinical Strategies Implementation Scale (CSI), an instrument designed to help providers measure the extent to which evidence-based strategies have been implemented in the treatment of persons with schizophrenia spectrum disorders. METHODS: Nine ordinal scales were devised to measure key aspects of treatment strategies that have been associated with clinical and social recovery from schizophrenia: goal- and problem-oriented assessment, medication strategies, assertive case management, mental health education, caregiver-based problem solving, living skills training, psychological strategies for residual problems, crisis prevention and intervention, and booster sessions. A study of interrater reliability was conducted with 15 trained raters from participating centers in Athens, Auckland, Bonn, Budapest, Gothenburg, and Tokyo who assessed 54 cases. Each treatment strategy was weighted according to its effect size in clinical trials. Correlation analyses were conducted to explore associations between the total CSI score and ratings of clinical, social, and caregiver outcomes each year over four years of continued treatment of 51 patients. RESULTS: Interrater reliability ranged from .93 to .99. Four annual total CSI ratings were significantly correlated with impairment, disability, functioning, work activity, and an index of recovery. Most correlations were stronger in years 3 and 4 than in years 1 and 2. CONCLUSIONS: Reliable and valid assessment of the implementation of evidence based strategies in clinical practice is feasible. The quality of integrated program implementation may be associated with improved clinical and social recovery from schizophrenic disorders. PMID- 16339623 TI - Commentary: fidelity measurement and the CSI. PMID- 16339624 TI - Commentary: fidelity measurement, the CSI, and the current evidence base. PMID- 16339625 TI - A comparison of service use and costs among adults with ADHD and adults with other chronic diseases. AB - OBJECTIVE: This study compared health service use and costs among adults with attention-deficit hyperactivity disorder (ADHD) and adults with depression, diabetes, or seasonal allergic rhinitis (seasonal allergy). METHODS: Pharmacy and medical claims data from January 1, 1999, to December 31, 2001, were obtained from a large U.S. managed care plan. Patients were divided into one of four mutually exclusive cohorts (ADHD, depression, diabetes, or seasonal allergy) on the basis of their diagnosis or prescriptions. Age, gender, comorbid conditions, and type of dominant provider were compared for the four groups. Thereafter, general linear models with post-hoc multiple comparison tests were used to compare health service use and costs by site of service. RESULTS: The study population included 143,561 patients. The four mutually exclusive groups included 58,017 with depression, 45,479 with diabetes, 33,272 with a seasonal allergy, and 6,793 had ADHD. Comorbid mental health conditions occurred with significantly greater frequency among patients with ADHD and among those with depression than among patients in the other groups. Patients with ADHD and those with depression received care from a mental health provider significantly more often than those with diabetes or a seasonal allergy. Patients with ADHD had fewer average outpatient, inpatient, and emergency department visits than patients with depression or diabetes, and office visit frequency for patients with ADHD was significantly different from that of patients with a seasonal allergy. Annual total costs were lowest among patients with a seasonal allergy ($2,743), which were not significantly different than the costs for patients with ADHD ($3,020). CONCLUSIONS: Adult ADHD poses an economic burden that is less than that of depression or diabetes but greater than that of a seasonal allergy. PMID- 16339626 TI - Disparities in mental health treatment in U.S. racial and ethnic minority groups: implications for psychiatrists. PMID- 16339627 TI - Commentary: APA's Efforts to Eliminate Disparities. PMID- 16339628 TI - Assessment of police calls for suicidal behavior in a concentrated urban setting. AB - As a result of deinstitutionalization over the past half-century, police have become frontline mental health care workers. This study assessed five-year patterns of police calls for suicidal behavior in Toronto, Canada. Police responded to an average of 1,422 calls for suicidal behavior per year, 15 percent of which involved completed suicides (24 percent of male callers and 8 percent of female callers). Calls for suicidal behavior increased by 4 percent among males and 17 percent among females over the study period. The rate of completed suicides decreased by 22 percent among males and 31 percent among females. Compared with women, men were more likely to die from physical (as opposed to chemical) methods (22 percent and 43 percent, respectively). The study results highlight the importance of understanding changes in patterns and types of suicidal behavior to police training and preparedness. PMID- 16339629 TI - A review of medication administration errors reported in a large psychiatric hospital in the United kingdom. AB - A retrospective analysis of reports of medication administration errors over a period of three and a half years was carried out in a UK psychiatric hospital. A total of 112 errors and "near misses" were studied. The reporting rate increased over time. Psychotropic, intramuscular, and as-needed medications were overrepresented in the error reports. Fifteen percent of the errors had the potential to cause moderate or severe harm to patients. The two most common factors cited by nurses as contributing to error causation were a busy, noisy environment and personal factors, such as feeling tired or unsupported. Physicians were cited as having contributed to some errors. PMID- 16339630 TI - Screening for infectious diseases among patients in a state psychiatric hospital. AB - This cross-sectional retrospective study examined the frequency of positive tests for infectious diseases among 655 patients who were admitted to a state psychiatric hospital from 1997 to 1999. Not all patients who were admitted to the hospital were tested for each disease. Rates of positive tuberculin (20.2 percent), hepatitis B (23.9 percent), and hepatitis C (21.5 percent) tests were all significantly greater than estimates for the U.S. general population. A minority of patients were tested for HIV, and 29.0 percent were positive. The rate of hepatitis A was similar to that in the general population (33.2 percent). Independent risk factors for markers of disease included age, immigrant status, homelessness, and history of substance use. These results confirm the importance of screening for infectious diseases among individuals with severe and persistent mental illness. PMID- 16339631 TI - A simulated workplace experience for nonmedicated adults with and without ADHD. AB - Eighteen nonmedicated adults with attention-deficit hyperactivity disorder (ADHD) and 18 who did not have ADHD were evaluated in a full-day simulated workplace experience. It was hypothesized that adults with ADHD would evidence greater impairments on simulated tasks, off-task behavior, and self-reported ADHD symptoms than those without ADHD. Participants were compared on self-reported ADHD symptoms, objective observations, and performance on written tasks. Significant differences were noted in reading comprehension and math fluency as well as observer-rated and self-reported behavior, but not attention. The results of this study suggest that ADHD among adults is associated with significant deficits in performance of workplace tasks, internal experiences, and external observations of core symptoms of ADHD. PMID- 16339632 TI - Past and current views on the use of seclusion and restraint in treatment. PMID- 16339633 TI - Recovering the momentum for federal support of multisite demonstrations. PMID- 16339634 TI - An emergency department's response to a patient's threat to kill the president. PMID- 16339636 TI - Safe and minimal use of seclusion and restraint. PMID- 16339637 TI - Oxcarbazepine and risk of falls among psychiatric inpatients. PMID- 16339638 TI - Family participation in the treatment of persons with serious mental illness. PMID- 16339639 TI - Meteorologic factors in emergency evaluation, admission, and discharge. PMID- 16339640 TI - IOM report focuses on the "quality chasm" in mental health and substance abuse treatment. PMID- 16339641 TI - Murine embryonic stem cells secrete cytokines/growth modulators that enhance cell survival/anti-apoptosis and stimulate colony formation of murine hematopoietic progenitor cells. AB - Stromal cell-derived factor (SDF)-1/CXCL12, released by murine embryonic stem (ES) cells, enhances survival, chemotaxis, and hematopoietic differentiation of murine ES cells. Conditioned medium (CM) from murine ES cells growing in the presence of leukemia inhibitory factor (LIF) was generated while the ES cells were in an undifferentiated Oct-4 expressing state. ES cell-CM enhanced survival of normal murine bone marrow myeloid progenitors (CFU-GM) subjected to delayed growth factor addition in vitro and decreased apoptosis of murine bone marrow c kit(+)lin- cells. ES CM contained interleukin (IL)-1alpha, IL-10, IL-11, macrophage-colony stimulating factor (CSF), oncostatin M, stem cell factor, vascular endothelial growth factor, as well as a number of chemokines and other proteins, some of which are known to enhance survival/anti-apoptosis of progenitors. Irradiation of ES cells enhanced release of some proteins and decreased release of others. IL-6, FGF-9, and TNF-alpha, not detected prior to irradiation was found after ES cells were irradiated. ES cell CM also stimulated CFU-GM colony formation. Thus, undifferentiated murine ES cells growing in the presence of LIF produce/release a number of biologically active interleukins, CSFs, chemokines, and other growth modulatory proteins, results which may be of physiological and/or practical significance. PMID- 16339642 TI - Local irradiation not only induces homing of human mesenchymal stem cells at exposed sites but promotes their widespread engraftment to multiple organs: a study of their quantitative distribution after irradiation damage. AB - Mesenchymal stem cells (MSCs) have been shown to migrate to various tissues. There is little information on the fate and potential therapeutic efficacy of the reinfusion of MSCs following total body irradiation (TBI). We addressed this question using human MSC (hMSCs) infused to nonobese diabetic/ severe combined immunodeficient (NOD/SCID) mice submitted to TBI. Further, we tested the impact of additional local irradiation (ALI) superimposed to TBI, as a model of accidental irradiation. NOD/SCID mice were transplanted with hM-SCs. Group 1 was not irradiated before receiving hMSC infusion. Group 2 received only TBI at a dose of 3.5 Gy, group 3 received local irradiation to the abdomen at a dose of 4.5 Gy in addition to TBI, and group 4 received local irradiation to the leg at 26.5 Gy in addition to TBI. Fifteen days after irradiation, quantitative and spatial distribution of the hMSCs were studied. Histological analysis of mouse tissues confirmed the presence of radio-induced lesions in the irradiated fields. Following their infusion into nonirradiated animals, hMSCs homed at a very low level to various tissues (lung, bone marrow, and muscles) and no significant engraftment was found in other organs. TBI induced an increase of engraftment levels of hMSCs in the brain, heart, bone marrow, and muscles. Abdominal irradiation (AI) as compared with leg irradiation (LI) increased hMSC engraftment in the exposed area (the gut, liver, and spleen). Hind LI as compared with AI increased hMSC engraftment in the exposed area (skin, quadriceps, and muscles). An increase of hMSC engraftment in organs outside the fields of the ALI was also observed. Conversely, following LI, hMSC engraftment was increased in the brain as compared with AI. This study shows that engraftment of hMSCs in NOD/ SCID mice with significantly increased in response to tissue injuries following TBI with or without ALI. ALI induced an increase of the level of engraftment at sites outside the local irradiation field, thus suggesting a distant (abscopal) effect of radiation damage. This work supports the use of MSCs to repair damaged normal tissues following accidental irradiation and possibly in patients submitted to radiotherapy. PMID- 16339643 TI - Early response of endogenous adult neural progenitor cells to acute spinal cord injury in mice. AB - Adult neural progenitor cells (NPCs) are an attractive source for functional replacement in neurodegenerative diseases and traumatic injury to the central nervous system (CNS). It has been shown that transplantation of neural stem cells or NPCs into the lesioned region partially restores CNS function. However, the capacity of endogenous NPCs in replacement of neuronal cell loss and functional recovery of spinal cord injury (SCI) is apparently poor. Furthermore, the temporal and spatial response of endogenous adult NPCs to SCI remains largely undefined. To this end, we have analyzed the early organization, distribution, and potential function of NPCs in response to SCI, using nestin enhancer (promoter) controlled LacZ reporter transgenic mice. We showed that there was an increase of NPC proliferation, migration, and neurogenesis in adult spinal cord after traumatic compression SCI. The proliferation of NPCs detected by 5 bromodeoxyuridine incorporation and LacZ staining was restricted to the ependymal zone (EZ) of the central canal. During acute SCI, NPCs in the EZ of the central canal migrated vigorously toward the dorsal direction, where the compression lesion is generated. The optimal NPC migration occurred in the adjacent region close to the epicenter. More significantly, there was an increased de novo neurogenesis from NPCs 24 hours after SCI. The enhanced proliferation, migration, and neurogenesis of (from) endogenous NPCs in the adult spinal cord in response to SCI suggest a potential role for NPCs in attempting to restore SCI-mediated neuronal dysfunction. PMID- 16339644 TI - In vitro-derived "neural stem cells" function as neural progenitors without the capacity for self-renewal. AB - Hematopoietic stem cells have been defined by their ability to self-renew and successfully reconstitute hematopoiesis throughout the life of a transplant recipient. Neural stem cells (NSCs) are believed to exist in the regenerating regions of the brain in adult mice: the subependymal zone (SEZ) of the lateral ventricles (LVs) and the hippocampal dentate gyrus. Cells from the SEZ can be cultured to generate neurospheres or multipotent astrocytic stem cells (MASCs), both of which demonstrate the stem cell qualities of multipotency and self renewal in vitro. Whether neurospheres and MASCs possess the true stem cell quality of functional self-renewal in vivo is unknown. The definitive tests for this unique capability are long-term engraftment and serial transplantation. Both neurospheres and MASCs transplanted into the LVs of C57BL/6 mice resulted in short-term engraftment into the recipient brain, with donor-derived migratory neuroblasts visible in the rostral migratory stream and olfactory bulb after transplantation. To test in vivo expansion/self-renewal of the transplanted cells, we attempted to reisolate donor-derived neurospheres and MASCs. Even when rigorous drug selection was used to select for rare events, no donor-derived neurospheres or MASCs could be reisolated. Furthermore, donor-derived migratory neuroblasts were not observed in the rostral migratory stream (RMS) for more than 1 month after transplantation, indicating a transient rather than long-term engraftment. Therefore, in vitro-derived neurospheres and MASCs do not function as NSCs with long-term, self-renewal capabilities in vivo but instead represent short-term neural progenitor cells as defined by an in vivo functional assay. PMID- 16339645 TI - Thalassemia: the continued challenge. AB - This overview describes the history of transfusion therapy and consequent iron overload in thalassemia. It emphasizes the importance of measurement of hepatic iron and reviews the history of chelation therapy. It briefly describes the discoveries of the genetic basis of thalassemia and the application of that knowledge in prenatal diagnosis. The review goes on to emphasize pharmaceutical efforts to induce fetal hemoglobin synthesis in thalassemic red cells and ends with a discussion of oral iron chelators, stem cell transplant, and the status of gene therapy. PMID- 16339646 TI - Keynote address: The challenge of thalassemia for the developing countries. AB - The thalassemias occur at some of their highest frequencies in the developing countries, particularly those of Asia. In many countries, facilities for the control of these conditions are extremely limited. Although a great deal can be done to help the situation by developing further North-South and South-South partnerships for disseminating better practice, the major problem for the future lies in the unwillingness of governments and international health agencies to accept that the thalassemias present a health burden comparable to that of other major diseases in the developing countries. However, preliminary analyses suggest that, at least in the case of Asia, this is not true. Further work of this nature, together with more detailed frequency and economic data, are required to provide solid evidence for the health burden posed by thalassemia in the developing world. Unless this is done, the large populations of patients with thalassemia in these countries will continue to be neglected. PMID- 16339647 TI - Changing patterns of thalassemia worldwide. AB - Thalassemia is a growing global public health problem with an estimated 900,000 births of clinically significant thalassemia disorders expected to occur in the next 20 years. This growth will occur in disorders previously uncommon in many parts of the world. In particular, hemoglobin (Hb) E-beta-thalassemia and Hb H disease account for much of the projected increases in thalassemia. Worldwide, Hb E-beta-thalassemia is one of the most frequent hemoglobinopathies. The incidence of Hb E approaches 60% of the populations in many regions of Southeast Asia. In coastal regions of North America, its prevalence is rapidly growing. The severity of Hb E-beta-thalassemia ranges from a complete lack of symptoms to transfusion dependence. alpha-Thalassemia diseases, often considered benign, are now recognized to be more severe than originally reported. Hb H, Hb H-Constant Spring (CS), and homozygous alpha-thalassemia affect at least a million people worldwide. California considers Hb H disease a public health problem and has initiated a neonatal screening program for Hb H and particularly Hb H-CS. Homozygous alpha-thalassemia, usually fatal, is also being more commonly detected. Several regions have initiated universal prenatal screening programs to address homozygous alpha-thalassemia. In summary, the prognosis for thalassemia disorders is improving, but prenatal diagnosis and neonatal screenings are needed. Comprehensive services that address language and social barriers as well as access to Hb F-enhancing agents and transfusions are needed. PMID- 16339648 TI - Alpha-thalassemia: Hb H disease and Hb Barts hydrops fetalis. AB - alpha-Thalassemia mutations are one of the most common mutations of man, and they cause Hb H disease and Hb Barts hydrops fetalis. Hb H disease is not necessarily a benign disorder as has been generally thought. Furthermore, in southern China and in Southeast Asia, there are 2-3 times more fetuses afflicted with the invariably fatal Hb Barts hydrops fetalis than with the beta-thalassemia major or intermedia. These findings underscore the public health importance of these hereditary disorders, and they call for better education, diagnosis, treatment, prevention, and research for these diseases. PMID- 16339649 TI - Hemoglobin E-beta-thalassemia: Progress report from the International Study Group. AB - A long-term observational study of Hb E-beta-thalassemia in Sri Lanka is beginning to define some of the genetic and environmental factors that are responsible for its remarkable phenotypic variability. In this population there is a very small difference between the steady-state hemoglobin levels between the mild and severe phenotypes, and it has been possible to stop transfusion in many of those who have been on long-term treatment of this kind. These preliminary observations, made over the last 7 years, provide directions for future research into this increasingly important disease. PMID- 16339650 TI - Survival and complications in thalassemia. AB - The life expectancy of patients with thalassemia major has significantly increased in recent years, as reported by several groups in different countries. However, complications are still frequent and affect the patients' quality of life. In a recent study from the United Kingdom, it was found that 50% of the patients had died before age 35. At that age, 65% of the patients from an Italian long-term study were still alive. Heart disease is responsible for more than half of the deaths. The prevalence of complications in Italian patients born after 1970 includes heart failure in 7%, hypogonadism in 55%, hypothyroidism in 11%, and diabetes in 6%. Similar data were reported in patients from the United States. In the Italian study, lower ferritin levels were associated with a lower probability of experiencing heart failure and with prolonged survival. Osteoporosis and osteopenia are common and affect virtually all patients. Hepatitis C virus antibodies are present in 85% of multitransfused Italian patients, 23% of patients in the United Kingdom, 35% in the United States, 34% in France, and 21% in India. Hepatocellular carcinoma can complicate the course of hepatitis. A survey of Italian centers has identified 23 such cases in patients with a thalassemia syndrome. In conclusion, rates of survival and complication free survival continue to improve, due to better treatment strategies. New complications are appearing in long-term survivors. Iron overload of the heart remains the main cause of morbidity and mortality. PMID- 16339651 TI - Role of intergenic human gamma-delta-globin sequences in human hemoglobin switching and reactivation of fetal hemoglobin in adult erythroid cells. AB - The details of the molecular events regulating normal human hemoglobin switching and reactivation of fetal hemoglobin in adult hematopoietic cells are unclear. The potential role of sequences between the human gamma- and delta-globin genes (intergenic gamma-delta sequences) in this process has been in question until the recent finding that two patients homozygous for the Corfu deletion, involving the loss of 7.2 kb of the intergenic gamma-delta region upstream of the delta gene, have 88% and 90% fetal hemoglobin, only mild anemia, and no transfusion requirements. These results provide the first strong evidence in humans that the gamma-delta intergenic sequences alone have a role in the reactivation of fetal hemoglobin in adult-type cells, and perhaps are involved in normal hemoglobin switching as well. The polypyrimidine (PYR) complex is a hematopoietic cell specific and stage-specific chromatin remodeling complex that binds upstream of the human delta-globin gene within the Corfu deletion. Deletion of the PYR binding site has been shown to delay human gamma-to-beta globin switching. The PYR complex is present in adult human hematopoietic cells and absent in fetal embryonic cells: properties of a globin-switching complex. Taken together, the data from patients with the Corfu deletion and the PYR complex results suggest that intergenic gamma-delta sequences are involved in human gamma-to-beta globin switching and reactivation of fetal hemoglobin in adult cells. PMID- 16339652 TI - Isolation and characterization of hematopoietic transcription factor complexes by in vivo biotinylation tagging and mass spectrometry. AB - We have described the application of a simple biotinylation tagging approach for the direct purification of tagged transcription factor complexes, based on the use of artificial short peptide tags that are specifically and efficiently biotinylated by the bacterial BirA biotin ligase, which is co-expressed in cells with the tagged factor. We used this approach to initially characterize complexes formed by the hematopoietic transcription factor GATA-1 in erythroid cells. GATA 1 is essential for the erythroid differentiation, its functions encompassing upregulation of erythroid genes, repression of alternative transcription programs, and suppression of cell proliferation. However, it was not clear how all of these GATA-1 functions are mediated. Our work describes, for the first time, distinct GATA-1 interactions with the essential hematopoietic factor Gfi 1b, the repressive MeCP1 complex, and the chromatin remodeling ACF/WCRF complex, in addition to the known GATA-1/FOG-1 and GATA-1/TAL-1 complexes. We also provide evidence that distinct GATA-1 complexes are associated with specific GATA-1 functions in erythroid differentiation, for example, GATA-1/Gfi-1b with the suppression of cell proliferation and GATA-1/FOG-1/MeCP1 with the repression of other hematopoietic transcription programs. We next applied the biotinylation tag to Ldb-1, a known partner of GATA-1, and characterized a number of novel interaction partners that are essential in erythroid development, in particular, Eto-2, Lmo4, and CdK9. Last, we are in the process of applying the same technology to characterize the factors that are bound to the suppressed gamma globin promoter in vivo. PMID- 16339653 TI - Germline epimutation: A basis for epigenetic disease in humans. AB - Epigenetic modifications of DNA produce reversible and clonally heritable alterations in transcription state. Errors in the elaborate apparatus of epigenetic silencing possessed by higher eukaryotes can lead to "epimutation," abnormal silencing of a gene. It was supposed that an epimutation in the germline would produce a phenotype equivalent to that resulting from an inactivating germline mutation in the same gene. In testing this hypothesis individuals were identified in whom one allele of the gene encoding the DNA mismatch repair protein MLH1 is epigenetically silenced throughout the soma (implying a germline event). These individuals fit the clinical criteria for hereditary nonpolyposis colorectal cancer, which is usually produced by germline mutation of MLH1. None of the affected individuals have any genetic abnormality that would explain the presence of the epimutation. Thus, an epimutation can phenocopy a genetic disease; this innate epigenetic defect is not necessarily the result of anything other than chance. Epigenetic phenomena tend to be stochastic, reversible, and mosaic; the occurrence and inheritance of epimutations are likely to have rules completely different from those of Mendelian genetics. The application of this principle to the thalassemias is discussed. PMID- 16339655 TI - Understanding alpha-globin gene regulation: Aiming to improve the management of thalassemia. AB - Over the past 50 years, many advances in our understanding of the general principles controlling gene expression during hematopoiesis have come from studying the synthesis of hemoglobin. Discovering how the alpha- and beta-globin genes are normally regulated and documenting the effects of inherited mutations that cause thalassemia have played a major role in establishing our current understanding of how genes are switched on or off in hematopoietic cells. Previously, nearly all mutations causing thalassemia have been found in or around the globin loci, but rare inherited and acquired trans-acting mutations are being found more often. Such mutations have demonstrated new mechanisms underlying human genetic disease. Furthermore, they are revealing new pathways in the regulation of globin gene expression that, in turn, may open up new avenues for improving the management of patients with common types of thalassemia. PMID- 16339654 TI - Progress toward the genetic treatment of the beta-thalassemias. AB - The beta-thalassemias are congenital anemias that are caused by mutations that reduce or abolish expression of the beta-globin gene. They can be cured by allogeneic hematopoietic stem cell (HSC) transplantation, but this therapeutic option is not available to most patients. The transfer of a regulated beta-globin gene in autologous HSCs is a highly attractive alternative treatment. This strategy, which is simple in principle, raises major challenges in terms of controlling expression of the globin transgene, which ideally should be erythroid specific, differentiation- and stage-restricted, elevated, position independent, and sustained over time. Using lentiviral vectors, May et al. demonstrated in 2000 that an optimized combination of proximal and distal transcriptional control elements permits lineage-specific and elevated beta-globin expression, resulting in therapeutic hemoglobin production and correction of anemia in beta-thalassemic mice. Several groups have by now replicated and extended these findings to various mouse models of severe hemoglobinopathies, thus fueling enthusiasm for a potential treatment of beta-thalassemia based on globin gene transfer. Current investigation focuses on safety issues and the need for improved vector production methodologies. The safe implementation of stem cell-based gene therapy requires the prevention of the formation of replication-competent viral genomes and minimization of the risk of insertional oncogenesis. Importantly, globin vectors, in which transcriptional activity is highly restricted, have a lesser risk of activating oncogenes in hematopoietic progenitors than non-tissue specific vectors, by virtue of their late-stage erythroid specificity. As such, they provide a general paradigm for improving vector safety in stem cell-based gene therapy. PMID- 16339656 TI - Role of alpha-hemoglobin-stabilizing protein in normal erythropoiesis and beta thalassemia. AB - Hemoglobin (Hb) synthesis is coordinated by homeostatic mechanisms to limit the accumulation of free alpha or beta subunits, which are cytotoxic. Alpha hemoglobin-stabilizing protein (AHSP) is an abundant erythroid protein that specifically binds free alphaHb, stabilizes its structure, and limits its ability to participate in chemical reactions that generate reactive oxygen species. Gene ablation studies in mice demonstrate that AHSP is required for normal erythropoiesis. AHSP-null erythrocytes are short-lived, contain Hb precipitates, and exhibit signs of oxidative damage. Loss of AHSP exacerbates beta-thalassemia in mice, indicating that altered AHSP expression or function could modify thalassemia phenotypes in humans, a topic that is beginning to be explored in clinical studies. We used biochemical, spectroscopic, and crystallographic methods to examine how AHSP stabilizes alphaHb. AHSP binds the G and H helices of alphaHb on a surface that largely overlaps with the alpha1-beta1 interface of HbA. This result explains previous findings that betaHb can competitively displace AHSP from alphaHb to form HbA tetramer. Remarkably, binding of AHSP to oxygenated alphaHb induces dramatic conformational changes and converts the heme bound iron to an oxidized hemichrome state in which all six coordinate positions are occupied. This structure limits the reactivity of heme iron, providing a mechanism by which AHSP stabilizes alphaHb. These findings suggest a biochemical pathway through which AHSP might participate in normal Hb synthesis and modulate the severity of thalassemias. Moreover, understanding how AHSP stabilizes alphaHb provides a theoretical basis for new strategies to inhibit the damaging effects of free alphaHb that accumulates in beta-thalassemia. PMID- 16339657 TI - Role of iron in inducing oxidative stress in thalassemia: Can it be prevented by inhibition of absorption and by antioxidants? AB - The pathophysiology of thalassemia is, to a certain extent, associated with the generation of labile iron in the pathological red blood cell (RBC). The appearance of such forms of iron at the inner and outer cell surfaces exposes the cell to conditions whereby the labile metal promotes the formation of reactive oxygen species (ROS) leading to cumulative cell damage. Another source of iron accumulation results from increased absorption due to decreased expression of hepcidin. The presence of labile plasma iron (LPI) was carried out using fluorescent probes in the FACS. RNA expression of hepcidin was measured in two models of thalassemic mice. Hepcidin expression was also measured in human hepatoma HepG2 cells following incubation with thalassemic sera. LPI was identified and could be quantitatively measured and correlated with other parameters of iron overload. Hepcidin expression was downregulated in the livers of thalassemic mice, in major more than in intermedia. Thalassemic sera down regulated hepcidin expression in HepG2 liver cells. A possible way to decrease iron absorption could be by modulating hepcidin expression pharmacologically, by gene therapy or by its administration. Treatment with combination of antioxidants such as N-acetylcysteine for proteins and vitamin E for lipids in addition to iron chelators could neutralize the deleterious effects of ROS and monitored by quantitation of LPI. PMID- 16339658 TI - Objectives and mechanism of iron chelation therapy. AB - Prevention of cardiac mortality is the most important beneficial effect of iron chelation therapy. Unfortunately, compliance with the rigorous requirements of daily subcutaneous deferoxamine (DFO) infusions is still a serious limiting factor in treatment success. The development of orally effective iron chelators such as deferiprone and ICL670 is intended to improve compliance. Although total iron excretion with deferiprone is somewhat less than with DFO, deferiprone may have a better cardioprotective effect than DFO due to deferiprone's ability to penetrate cell membranes. Recent clinical studies indicate that oral ICL670 treatment is well tolerated and is as effective as parenteral DFO used at the standard dose of 40 mg/kg of body weight/day. Thus, for the patient with transfusional iron overload in whom results of DFO treatment are unsatisfactory, several orally effective agents are now available to avoid serious organ damage. Finally, combined chelation treatment is emerging as a reasonable alternative to chelator monotherapy. Combining a weak chelator that has a better ability to penetrate cells with a stronger chelator that penetrates cells poorly but has a more efficient urinary excretion may result in improved therapeutic effect through iron shuttling between the two compounds. The efficacy of combined chelation treatment is additive and offers an increased likelihood of success in patients previously failing DFO or deferiprone monotherapy. PMID- 16339659 TI - Ferritin as an iron concentrator and chelator target. AB - Ferritin has a broad role to play in new strategies for managing Cooley's anemia and the thalassemias. Serum ferritin iron content is relegated to reporting on tissue iron concentrations. Recently, a new property of ferritin was discovered: gated pores, which are highly conserved in ferritins of humans down to bacteria, and control iron flow to chelators. The pore gates can be selectively opened to increase chelator access by mutation, temperature, and physiological concentrations of urea. In another recent observation, the iron in ferritin from seeds such as soybeans has been shown to be readily available to tissues with high demand for iron, such as red blood cells, but slower to be mobilized in other tissues, compared to nonheme iron salts, presumably through a controlled iron gating mechanism. Because the iron pore gating property of ferritin is more thoroughly investigated, and the knowledge that much of the iron to be chelated in the thalassemias is from a solid iron mineral inside the ferritin protein nanocage, a new role of ferritin in regulating cellular iron homeostasis is established. Two new areas, based on recent knowledge of the molecular properties of ferritin, are (1) exploration of food ferritin as a potentially safer form of dietary nonheme iron, and (2) development of chelators targeted to ferritin protein pores that control chelator access. PMID- 16339660 TI - The design of orally active iron chelators. AB - It is now generally accepted that it is not possible to design iron(III) selective hexadentate chelators with high oral bioavailability. In order to achieve suitable levels of oral activity for the treatment of systemic iron overload either tridentate or bidentate molecules need to be investigated. There are a number of such molecules in clinical practice, including hydroxypyridin-4 ones, desferrithiocin analogues and bis-hydroxyphenyltriazoles. The underlying chemistry of each group is described, together with an indication of the distribution properties, redox cycling activity, and iron scavenging activity. PMID- 16339661 TI - Recent insights into interactions of deferoxamine with cellular and plasma iron pools: Implications for clinical use. AB - Despite the availability of deferoxamine (DFO) for more than three decades, its rates of interaction with cellular iron pools in different tissues, and the effects of its pharmacokinetics on the interaction with plasma iron pools, remain incompletely understood. The positive charge of DFO, together with the negative resting potential in vertebrate cells, favors cellular uptake, whereas the low lipophilicity and high molecular weight counter this effect. The findings presented suggest a facilitated uptake of DFO into hepatocytes, being several hundred-fold faster than into red cells. Antibodies that selectively recognize ferrioxamine (FO) show that initial hepatocellular iron chelation is cytosolic, but later transposes to lysosomal and ultimately canalicular compartments. Strong FO staining is visible in myocytes within 4-8 h after commencing a subcutaneous DFO infusion, indicating effective chelation of myocyte iron. A methodology was developed to study the interaction of DFO and its metabolites with plasma iron pools by stabilizing DFO with aluminum ions, thereby preventing iron shuttling from non-transferrin-bound iron (NTBI) onto DFO after plasma collection. DFO removes only about a third of NTBI rapidly, and NTBI is rarely cleared completely. Increasing DFO dosing does not increase NTBI removal, but instead leads to a greater rebound in NTBI on cessation of intravenous infusion. Thus, intermittent infusions of high-dose DFO are less desirable than continuous infusions at low doses, particularly in high-risk patients. Here the benefits of continuous DFO on heart function occur before changes in T2*-visible storage iron, consistent with early removal of a toxic labile iron pool within myocytes. PMID- 16339662 TI - Deferiprone: New insight. AB - Recent results from independent studies suggest that deferiprone is more cardioprotective than deferoxamine. Patients on long-term treatment with deferiprone have a better myocardial magnetic resonance imaging pattern and less chance to develop a new cardiac disease or worsen an existing one. Most of these observations are retrospective and require confirmation from randomized controlled trials. Other new observations regard the effects of combining the two chelators. Most results indicate an additional effect on iron excretion and a significant reduction of the time required to mitigate severe iron overload and to reverse clinical heart disease. Again, these data require confirmation, as they were mostly obtained on individual cases or small groups of patients treated with a wide range of combinations of the two chelators, but the univocity of results is impressive. After many years of controversy, deferiprone is emerging as a useful oral iron chelator that enhances the chances for the patient to have optimal treatment. Well-designed and -conducted studies will help in answering the questions still open. PMID- 16339663 TI - Combined therapy with deferoxamine and deferiprone. AB - Therapy with either deferiprone (DFP) or deferoxamine (DFO) is inadequate in achieving negative iron balance in many patients with thalassemia. There are mounting theoretical, experimental, and clinical evidences of increased efficacy when therapy includes both chelating agents. DFP and DFO chelate excess iron in different ways without affecting each other's metabolism. When both chelators are administered simultaneously, they interact either in an additive or synergistic manner, probably through "shuttling" iron from DFP to DFO. Iron-balance studies have shown that the use of both agents on the same day can induce negative iron balance in all patients. Long-term combined therapy with DFO with DFP results in considerable reduction of both ferritin levels and liver iron concentration as well as significant improvement in cardiac siderosis and function. This therapeutic regimen is well tolerated and safe, even though it may be related to a small increase in the incidence of agranulocytosis compared with DFP monotherapy. Apart from using both agents simultaneously, sequential administration of DFP and DFO has also shown promising results. Combining the available iron chelators offers many therapeutic options that can be tailored to each patient individually. It is an exciting advance in treating hemosiderosis in thalassemic patients. PMID- 16339664 TI - Evaluation of ICL670, a once-daily oral iron chelator in a phase III clinical trial of beta-thalassemia patients with transfusional iron overload. AB - Osteoporosis and osteopenia are frequent complications of thalassemia major (TM) and intermedia (TI). Osteoporosis was found in 23/25 patients with TI and in 115/239 patients with TM. In TM, no association was found with specific polymorphisms in candidate genes (vitamin D receptor, estrogen receptor, calcitonin receptor, and collagen type 1 alpha 1). Osteoporosis in TM female was strongly associated with primary amenorrhea (P < .0001), while in male patients with TM hypogonadism was not significantly related to BMD (P = .0001). Low BMD was also associated with cardiomiopathy (P = .01), diabetes mellitus (P = .0001), chronic hepatitis (P = .0029), and increased ALT (P = .01). PMID- 16339665 TI - Unrelated bone marrow transplantation for beta-thalassemia patients: The experience of the Italian Bone Marrow Transplant Group. AB - Bone marrow transplantation (BMT) remains the only potentially curative treatment for patients with thalassemia major. However, most candidates for BMT do not have a suitable family donor. In order to evaluate whether BMT from an HLA-matched unrelated volunteer donor can offer a probability of cure comparable to that obtained when the donor is a compatible sibling, we carried out a study involving 68 thalassemia patients transplanted in six Italian BMT Centers. Thirty-three males and 35 females (age range, 2-37 years; median age, 15) were transplanted from unrelated volunteer donors, all selected using high-resolution molecular typing of both HLA class I and II loci. Fourteen patients were classified in risk class 1; 16 in risk class 2; and 38 in risk class III of the Pesaro classification system. Nine patients (13%) had either primary or secondary graft failure. Fourteen patients (20%) died from transplant-related causes. Grade II-IV acute graft-versus-host disease (GVHD) developed in 24 cases (40%), and chronic GVHD in 10 cases (18%). Overall survival (OS) in the cohort of 68 patients was 79.3% (CI 67-88%), whereas the Kaplan-Meier estimates of disease-free survival (DFS) with transfusion independence was 65.8% (CI 54-77%). In the group of 30 thalassemic patients in risk classes 1 and 2, the probability of OS and DFS were 96.7% (CI 90-100%) and 80.0% (CI 65-94%), respectively, whereas in the 38 patients in class 3 OS was 65.2% (CI 49-80%) and DFS was 54.5% (CI 38-70%). These data show that when donor selection is based on stringent compatibility criteria, the results of unrelated transplantation in thalassemia patients are comparable to those obtained when the donor is a compatible sibling. PMID- 16339666 TI - Bone marrow transplantation in adults with thalassemia: Treatment and long-term follow-up. AB - Current regular blood transfusion programs and chelation treatment have considerably improved survival of patients with thalassemia, which resulted in a larger proportion of adult patients. However, disease- and treatment-related complications in these patients progress over time, causing severe morbidity and shortened life expectancy. Stem cell transplantation still remains the only cure currently available for patients with thalassemia. This study updates transplant outcomes in 107 adult patients with median age of 22 years (range, 17-35 years) who received bone marrow transplantation (BMT) from human leukocyte antigen (HLA) identical related donors between 1988 and 1996 (group A) and describes the results of BMT in 15 adult patients with median age of 21 years (range, 17-31 years) who were treated with a new treatment protocol (Protocol 26) between 1997 and 2003 (group B). The probability of survival, event-free survival, nonrejection mortality, and rejection for group A patients were 66%, 62%, 37%, and 4%, respectively, with a median follow-up of 12 years (range, 8.3-16.2 years). Group B patients treated with the new protocol had some improvement in thalassemia-free survival (67%) and lower transplant-related mortality (27%) than that of previous protocols. However, transplant-related mortality in these high risk patients remains elevated. Current myeloablative BMT in adult patients is characterized by higher transplant-related toxicity due to an advanced phase of disease. Although this new approach to transplant adult patients with a reduced dose intensity-conditioning regimen has improved thalassemia-free survival, transplant-related mortality in these high-risk patients remains elevated. PMID- 16339667 TI - Sibling donor cord blood transplantation for thalassemia major: Experience of the Sibling Donor Cord Blood Program. AB - The Sibling Donor Cord Blood (SDCB) Program was initiated in 1998 as a resource to collect, characterize, and release cord blood units (CBUs) from families affected by malignant and nonmalignant disorders for transplantation. Families in the United States were recruited by telephone after referrals by community and academic physicians. Collection kits were mailed to prospective participants and family members were instructed about CBU procurement from community hospitals and shipping to a central laboratory. Data about the infant's delivery and CBU harvest, CBU processing, prethaw characteristics, sterility, and human leukocyte antigen (HLA) typing were collected. Standard outcome data were collected after CBU release for transplantation. Descriptive analyses of CBU collections, processing, release, and transplantation outcomes were performed. Currently, 1617 CBU collections have been processed from families with thalassemia (6%), sickle cell disease (28%), malignant disorders (49%), and other rare hematological disorders (17%). Thirty-two of 96 donor-recipient pairs with thalassemia major were HLA identical and 14 have received cord blood transplantation, either alone or in combination with bone marrow or peripheral blood progenitor cells (N = 4) from the same donor. Eleven of the 14 survive free of thalassemia after transplantation. These preliminary results confirm the feasibility and utility of remote-site sibling donor cord blood collection and subsequent transplantation for hematological disorders, with a very high rate of usage from a cord blood bank dedicated to performing these unique collections. It was concluded that cord blood transplantation from sibling donors represents a suitable alternative to bone marrow transplantation. PMID- 16339668 TI - Stem cell transplantation with S-59 photochemically treated T-cell add-backs to establish allochimerism in murine thalassemia. AB - Hematopoietic cell transplantation (HCT) from HLA-identical sibling donors has curative potential for beta-thalassemia. The probability of surviving free of thalassemia under these conditions is approximately 85%. The application of this therapy is limited because many patients lack an HLA-identical sibling donor. HLA mismatched stem cell transplantation for thalassemia is severely restricted by graft rejection and the risks for graft-versus-host disease (GVHD). Thus, the development of a novel method that facilitates immunological tolerance and improves the safety of HLA-mismatched HCT would greatly expand the opportunity of HCT for thalassemia patients. We hypothesized that removal of T cells from the donor hematopoietic stem cell preparation and subsequent add-back after photochemical treatment with S-59, a psoralen, would promote and stabilize the engraftment and significantly reduce the risk of GVHD. This was tested in a MHC mismatched HCT model of murine thalassemia. S-59-treated T cells were infused simultaneously with bone marrow-derived stem cells into mice with a heterozygous deletion of one beta-globin alleles that had been conditioned with a sublethal dose of total body irradiation. Mice that received treated T cells showed increased engraftment compared to those that did not receive T cells. T-cell treatment improved survival without GVHD compared to recipients that received untreated T cells. We conclude that photochemical treatment of T cells facilitates engraftment and minimizes GVHD in allo-HCT for murine thalassemia, and sets the stage for further development of such protocols for the treatment of patients with thalassemia. PMID- 16339669 TI - Preimplantation genetics: Improving access to stem cell therapy. AB - There has been progress in the application of stem cell transplantation for treatment of an increasing number of severe congenital and acquired bone marrow disorders, currently restricted by the availability of human leukocyte antigen (HLA)-matched related donors. Preimplantation HLA typing has recently been introduced to improve the access to stem cell therapy for inherited bone marrow failures. Preimplantation genetic diagnosis (PGD) provides an option not only for avoiding an affected pregnancy with thalassemia and other inherited disorders but also for preselection of the HLA-compatible donors for affected siblings. Multiple short tandem repeat markers throughout the HLA region are applied for this purpose, allowing 100% accuracy of HLA typing, through picking up possible recombination in the HLA region, as well as the copy number of chromosome 6, which affect accuracy of preimplantation HLA typing. Present experience of preimplantation HLA typing includes preimplantation HLA typing in 180 cycles, 122 of which were done as part of PGD for Fanconi anemia, thalassemia, Wiscott Aldrich syndrome, hyper-immunoglobulin M syndrome, hypohidrotic ectodermal dysplasia with immune deficiency, and X-linked adrenoleukodystrophy, and 58 for the sole purpose of HLA typing for leukemias and for aplastic and Diamond Blackfan anemia. The applied method resulted in the accurate preselection and transfer of 100% HLA-matched embryos, yielding already three dozen clinical pregnancies and the birth of two dozen HLA-matched children to the siblings requiring stem cell transplantation. Successful therapy with HLA-matched stem cells, obtained from these PGD children, has been achieved already for Diamond Blackfan anemia hypohidrotic ectodermal dysplasia with immune deficiency and thalassemia. PMID- 16339670 TI - Pharmacological induction of fetal hemoglobin: Why haven't we been more successful in thalassemia? AB - The first studies of the pharmacological induction of fetal hemoglobin were conducted in patients with sickle cell disease and thalassemia. Although hydroxyurea was approved by the FDA for the treatment of sickle cell disease in 1996, no similar pharmacological agent(s) has been approved for the treatment of patients with thalassemic disorders. The small-scale studies of the induction of fetal hemoglobin in thalassemia have been generally disappointing. The aim of this report is to provide a critical analysis of the factors that may be responsible for our failure to develop an effective fetal hemoglobin induction therapy for patients with thalassemia. We also describe several areas for future investigation that may be critically important for the development of an effective therapy for thalassemia. PMID- 16339671 TI - Successful correction of the human Cooley's anemia beta-thalassemia major phenotype using a lentiviral vector flanked by the chicken hypersensitive site 4 chromatin insulator. AB - beta-Thalassemias are the most common single-gene disorders and are potentially amenable to gene therapy. While retroviral vectors carrying the human beta-globin cassette were notoriously unstable and expressed poorly, considerable progress has now been made using lentiviral vectors (LVs), which stably transmit the beta globin expression cassette. Mouse studies using LVs have shown correction of the beta-thalassemia-intermedia phenotype and a partial, variable correction of the mouse beta-thalassemia major phenotype, despite the use of beta-globin hypersensitive sites that are known to result in position-independent effects. Our group used the alpha-globin-hypersensitive site in self-inactivating (SIN) LVs with long-term expression in secondary mice that resisted methylation associated proviral silencing. However, these vectors also suffered from chromatin position effects. We therefore flanked a SIN-lentiviral vector carrying the human beta-globin expression cassette with a chromatin insulator and studied expression in bone marrow from four patients with transfusion-dependent human thalassemia major. We demonstrated normal levels of human beta-globin expression in erythroid cells produced in in vitro cultures for unilineage erythroid differentiation. There was restoration of effective erythropoiesis and reversal of the abnormally elevated apoptosis that characterizes beta-thalassemia. The gene-corrected human beta-thalassemia progenitor cells were transplanted into immune-deficient mice, where they underwent normal erythroid differentiation, expressed normal levels of human beta-globin, and displayed normal effective erythropoiesis 3-4 months after xenotransplantation. Variability of beta-globin expression in erythroid colonies derived in vitro or from xenograft bone marrow was similar to that seen in normal control subjects. Results show genetic correction of primitive human progenitor cells and normalization of the human thalassemia major phenotype. PMID- 16339672 TI - Single and combination drug therapy for fetal hemoglobin augmentation in hemoglobin E-beta 0-thalassemia: Considerations for treatment. AB - Patients with hemoglobin E (Hb E)-beta 0-thalassemia, one of the most common hemoglobinopathies worldwide, could benefit from drugs that increase fetal and total hemoglobin levels and thereby decrease the need for transfusions. The long term clinical outcome of such therapy, its hematologic effects, and which patients are likely to benefit from treatment are unknown. Consequently, the use of such drugs for Hb E-beta 0-thalassemia is limited, and countries where resources for safe and regular transfusion are scarce cannot benefit from them. In a multicenter trial of 42 patients treated with hydroxyurea for two years, almost half the patients demonstrated a significant increase in steady-state hemoglobin level. Drug toxicity was minimal. Combined treatment of hydroxyurea with erythropoietin benefited selected patients, but the addition of sodium phenyl butyrate was ineffective. After 5 years of follow-up, a subset of patients remained off transfusions. Hydroxyurea should be considered for a subset of Hb E beta 0-thalassemia patients. PMID- 16339673 TI - Induction of fetal globin in beta-thalassemia: Cellular obstacles and molecular progress. AB - Accelerated apoptosis of erythroid progenitors in beta-thalassemia is a significant barrier to definitive therapy because the beneficial effects of fetal globin-inducing agents on globin chain balance may not be inducible in cells in which programmed cell death is established early. Accordingly, our objectives have been to identify methods to decrease cellular apoptosis and to identify orally tolerable fetal globin gene inducers. A pilot clinical trial was conducted to determine whether combined use of a fetal globin gene inducer (butyrate) and rhu-erythropoietin (EPO), the hematopoietic growth factor that prolongs erythroid cell survival and stimulates erythroid proliferation, would produce additive hematologic responses in any thalassemia subjects. Butyrate and EPO were administered in 10 patients. Novel fetal globin gene inducers that also stimulate erythroid proliferation were evaluated for pharmacokinetic profiles. Patients with beta+-thalassemia had relatively low levels of endogenous EPO (<145 mU/mL) and had additive responses to administered EPO and butyrate. Patients with at least one beta 0-globin mutation had higher baseline HbF levels (>60%) and EPO levels (>160 mU/mL), and three-fourths of these subjects responded to the fetal globin gene inducer alone. A few select fetal globin-inducing short-chain fatty acid derivatives that stimulated cell proliferation also had favorable pharmacokinetics. These studies identify a significant subset of thalassemia patients who appear to require exogenous EPO to respond optimally to any HbF inducer, as well as new therapeutic candidates that act on both cellular and molecular pathologies of beta-thalassemia. Both approaches now offer excellent potential for tolerable, definitive treatment of beta-thalassemia. PMID- 16339674 TI - Caring for adults with thalassemia in a pediatric world. AB - Improved technology and medical advances have increased the life span for patients with thalassemia. Therefore, serious consideration must now be given to adult issues such as fertility, employment opportunities, medical insurance, and long-term coping with chronic illness. Since thalassemia is a childhood illness, most adults are seen in pediatric hospitals-often, in centers with a specialty for thalassemia. Compared to a decade ago, many more patients in thalassemia centers are adolescents or older. Unfortunately, pediatric hospitals are not fully equipped to meet the changing and complex needs of adults. Emergency room care, hospital admissions, decentralized care, comprehensive care, and psychosocial issues are current challenges that must be addressed. In this study, six adult patients were asked to keep track of their care for one month to further examine self-care for thalassemia, a high-maintenance disease. From a qualitative perspective, the issues and challenges that adults face are examined and solutions for improved care are discussed. PMID- 16339675 TI - Quality of life in thalassemia. AB - Morbidity and mortality related to thalassemia have been reduced significantly with modern medical treatment, and quality of life (QOL) should now be considered an important index of effective health care. An assessment of QOL differs from other forms of medical assessment in that it focuses on the individuals' own views of their well-being and assesses other aspects of life, giving a more holistic view of well-being. There is very little published work on evaluation of QOL in thalassemia. A suitable tool should be reproducible, sensitive to the major features of the condition that affect patients' lives, and applicable in the range of different cultural, age, and social settings. Such an instrument would be valuable in evaluating new forms of treatment and in comparing health outcomes between different clinics. Two instruments have been assessed, one derived from the WHOQOL-100 questionnaire, and one designed specifically for thalassemia, which assesses psychosocial and clinical burden, as they affect adult patients, parents, and children. Further studies are required to develop and assess such tools for use in thalassemia. Another approach is to seek patients' own views of their routine treatment and the extent to which medical treatment affects QOL. Results from patient questionnaires in the United Kingdom and Cyprus are consistent in finding problems with organization of transfusions, insufficient options with chelation therapy, and poor communication. Practical measures could be taken to address these issues. PMID- 16339676 TI - Thalassemia and learning: Neurocognitive functioning in children. AB - As more effective management and even cure of thalassemia become possible, attention is beginning to be directed to the potential neurologic and resulting neurocognitive effects of this illness on adults and children. Recent studies indicate that for adults with beta-thalassemia major and intermedia, and for children with sickle beta-thalassemia (Sbeta-thalassemia), there is a substantial risk for silent brain infarcts that may be associated with neurocognitive impairment similar to that reported for children with sickle cell anemia. Here the available literature in this area is reviewed and the limited outcomes are compared with those available from large, multicenter longitudinal studies of sickle cell anemia. On the basis of these comparisons, it is recommend that children with thalassemia be screened for specific neuropsychological impairments and that they be provided early intervention and special education access as available under the Individuals with Disabilities Education Act (IDEA) or the 504 Regulations of the Rehabilitation Act of 1973. PMID- 16339677 TI - Treatment of hepatitis C virus infection in thalassemia. AB - Treatment of hepatitis C virus (HCV) in the general population has improved over the last decade. Patients treated with peginterferon alfa (PegIFN) and ribavirin (RBV) combination therapy demonstrate overall 50-55% sustained viral response (SVR) with rates as high as 80% in patients with genotypes 2 and 3. Because RBV induces hemolysis and subsequently increases blood transfusion requirements, combination therapy has been considered contraindicated for hemoglobinopathies. This report reviews the response to interferon alfa and RBV (IFN/RBV) and PegIFN/RBV combination therapies in patients treated in the Northern California Comprehensive Thalassemia Center. A total of six thalassemia major patients were treated with IFN/RBV (n = 5; age: 4-38 years) or with PegIFN/RBV (n = 1; age: 26 years). Quantitative HCV RNA polymerase chain reaction and liver iron level assessment were completed. Transfusion volumes were obtained from patients' medical records. On IFN/RBV combination, four of five patients demonstrated SVR. The one patient on PegIFN/RBV showed end-treatment viral response after 6 months of therapy (genotype 3), but subsequently relapsed. Liver iron pretreatment level ranged from 0.2 to 22 mg/g dry weight, with a mean +/- SD of 7.9 +/- 7.7. Transfusion requirement increased by a median of 43.5% (range: 32-137%). Five of the six patients had liver iron measurements within 1 year following completion of treatment, with quantitative liver iron increasing in two patients by 2.5 mg/g dry weight, decreasing in two patients by 3 and 14 mg/g dry weight, and remaining unchanged in one patient. All patients were able to complete combination therapy, although dose reductions were required. Patients with thalassemia and high iron overload can obtain SVR after combination therapy with rates similar to those in the general population and without significant complications. Although transfusion requirements increased in most patients, iron burden was not necessarily increased. PMID- 16339678 TI - Fertility and pregnancy in thalassemia major. AB - Over the last 15 years, 22 women with thalassemia major have completed 29 pregnancies at the Royal Hospital in London. The major pre-pregnancy issues, medications, and pregnancy care are reviewed. Experience suggests that, with proper care and guidance, pregnancies among women with thalassemia major are practical and can have successful outcomes. PMID- 16339679 TI - A phase I/II clinical trial of beta-globin gene therapy for beta-thalassemia. AB - Recent success in the long-term correction of mouse models of human beta thalassemia and sickle cell anemia by lentiviral vectors and evidence of high gene transfer and expression in transduced human hematopoietic cells have led to a first clinical trial of gene therapy for the disease. A LentiGlobin vector containing a beta-globin gene (beta(A-T87Q)) that produces a hemoglobin (Hbbeta(A T87Q)) that can be distinguished from normal hemoglobin will be used. The LentiGlobin vector is self-inactivating and contains large elements of the beta globin locus control region as well as chromatin insulators and other features that should prevent untoward events. The study will be done in Paris with Eliane Gluckman as the principal investigator and Philippe Leboulch as scientific director. PMID- 16339680 TI - Coagulation and splenectomy: an overview. AB - Venous thromboembolic events, such as pulmonary embolism, deep venous thrombosis, and portal vein thrombosis, have been observed in adult thalassemia patients, mainly in beta-thalassemia intermedia. The clinical findings are consistent with the observation of several alterations that indicate a state of activation of the hemostatic mechanisms in thalassemias. These alterations have usually been related to high platelet counts due to splenectomy and/or liver dysfunction. In a retrospective study of a large cohort of adults with thalassemia, we found a larger prevalence of venous thromboembolic events in transfusion-independent patients with thalassemia intermedia (29%) than in regularly transfused patients with thalassemia major (2%); moreover, the higher prevalence occurred particularly in splenectomized thalassemia intermedia patients. More recently, a multicenter study involving 56 tertiary referral centers in 7 countries was planned to assess the magnitude of thrombotic risk in thalassemia patients. The total number of patients who had thrombotic events was 146 (1.65%) out of 8860, with a prevalence of 0.9% in thalassemia major and 4% in thalassemia intermedia. The highest prevalence was confirmed in splenectomized patients. The observation that thrombotic events are more frequent in beta-thalassemia patients who are not receiving regular transfusions (thalassemia intermedia or thalassemia major patients in less developed countries with limited transfusion resources) or in thalassemic patients who have undergone splenectomy strongly supports the procoagulant activity of circulating damaged red blood cells. PMID- 16339681 TI - Transfusion safety: Where are we today? AB - Over the past decades, the incidence of clinically significant transfusion transmitted diseases has been dramatically reduced. These reductions have occurred because of a multifocal approach to the collection, processing, and release of blood and blood components. Research has focused on characterizing specific pathogens and their infectious patterns, especially the early viremic phase. Donor motivation and characteristics have been investigated, and restrictive eligibility criteria have been established. Regulatory oversight has been strengthened in the United States and hemovigilance systems established in many countries of the European Union, Canada, and Japan to identify new and emerging infectious and noninfectious transfusion risks. Such systems are required because many emerging pathogens will elude the stringent and sensitive donor testing already in place which, unfortunately, requires advanced technologies. Despite the remarkable progress, the processes of delivering a transfusion to a patient provide additional opportunity for risk, but have been less- well studied and the solutions have been less-well defined. Specific areas of concern include the collection and correct identification of patient samples, selection of modified or unique products, blood administration process, and establishment of specific indications for transfusion. In this chapter, we review the status of transfusion safety in the United States and discuss some emerging infectious and noninfectious risks of transfusion that will become future areas for basic and translational research. The disparities that exist in blood safety in developing countries' red cell alloimmunization and autoimmunity will not be covered. PMID- 16339682 TI - Pulmonary hypertension in beta-thalassemia. AB - Cardiac involvement represents the leading cause of mortality in both forms of beta-thalassemia, namely, thalassemia major (TM) and thalassemia intermedia (TI), and pulmonary hypertension (PHT) is part of the cardiopulmonary complications of the disease. PHT was initially documented in a small group of TI patients with right heart failure. In a subsequent study of a large 110-patient series, aged 32.5 +/- 11.4 years, age-related PHT was encountered in nearly 60% of cases, having caused right heart failure in six of them; interestingly, all patients had preserved left ventricular systolic function. Conflicted evidence, however, existed with respect to the development of PHT in heterogeneously treated and young TM populations. To resolve this discrepancy, a recent study compared cardiac disease between two large aged-matched groups of TM (n = 131) and TI (n = 74) patients, both treated uniformly in the currently accepted manner (regular transfusion and chelation therapy in TM, absence of any particular treatment in TI); well-treated TM patients, in contrast to TI patients, did not develop PHT, while systolic left ventricular dysfunction was present only in TM cases. PHT in beta-thalassemia results from a rather complex pathophysiology, in which chronic tissue hypoxia seems to hold a key role. Although both forms of the disease share a common molecular background, the diverse severity of the genetic defect and of the resulting clinical phenotype require a different therapeutic approach. Regular lifelong therapy in TM patients eliminates chronic hypoxia, thereby preventing PHT, whereas the absence of systematic treatment in TI leads to a cascade of reactions that compensate for chronic anemia, but at the same time allow the development of PHT. PMID- 16339683 TI - Monitoring long-term efficacy of iron chelation treatment with biomagnetic liver susceptometry. AB - In patients with thalassemia, the assessment of liver iron concentration (LIC) can be used to initiate chelation treatment with desferrioxamine (DFO), deferiprone (DFP), or novel chelators (deferasirox); to adjust chelation dose according to the actual blood transfusion rate; and to monitor chelation efficacy. The results from measurements by SQUID biomagnetic liver susceptometry in the LIC range 17-11,500 microg/g of liver in about 1000 patients were used to derive nonstandard parameters, which may be useful in the treatment monitoring of patients with thalassemia. From these measurements, including liver volumes, the documented chelation dose rates, and the blood transfusion rates, the chelator index (equivalent Therapeutical Index), the total body iron elimination rate, and the molar efficacy were calculated. Chelator indices (CIs) ranged from 0.1 to 11.7 mmol/d/g of Fe for DFO, with a threshold of CI greater than 1.2 mmol/d/g of Fe indicating DFO toxicity. For DFP, CI ranged from 0.1 to 23.2 mmol/d/g of Fe. In long-term studies (2 and 4 years), mean molar efficacies of DFO and DFP were found to be quite stable with 17.6 +/- 4.8% and 4.9 +/- 1.4%, respectively. Currently, specific chelation dose is based upon body weight. Because liver iron measurements by biosusceptometry are now regularly available in Europe and America, as well as quantitative MRI worldwide, these methods may be used to adjust chelation treatment regimens to body iron stores. PMID- 16339684 TI - Methods for noninvasive measurement of tissue iron in Cooley's anemia. AB - To examine the relationship between myocardial storage iron and body iron burden, as assessed by hepatic storage iron measurements, we studied 22 patients with transfusion-dependent thalassemia syndromes, all being treated with subcutaneous deferoxamine, and 6 healthy subjects. Study participants were examined with a Philips 1.5-T Intera scanner using three multiecho spin echo sequences with electrocardiographic triggering and respiratory navigator gating. Myocardial and hepatic storage iron concentrations were determined using a new magnetic resonance method that estimates total tissue iron stores by separately measuring the two principal forms of storage iron, ferritin and hemosiderin. In a subset of 10 patients with beta-thalassemia major, the hepatic storage iron concentration had been monitored repeatedly for 12-14 years by chemical analysis of tissue obtained by liver biopsy and by magnetic susceptometry. In this subset, we examine the relationship between hepatic iron concentration over time and our current magnetic resonance estimates of myocardial iron stores. No significant relationship was found between simultaneous estimates of myocardial and hepatic storage iron concentrations. By contrast, in the subset of 10 patients with beta thalassemia major, the correlation between the 5-year average of hepatic iron concentration and the current myocardial storage iron was significant (R = .67, P = .03). In these patients, myocardial storage iron concentrations seem to reflect the control of body iron over a period of years. Magnetic resonance methods promise to provide more effective monitoring of iron deposition in vulnerable tissues, including the liver, heart, and endocrine organs, and could contribute to the development of iron-chelating regimens that more effectively prevent iron toxicity. PMID- 16339685 TI - T2* magnetic resonance and myocardial iron in thalassemia. AB - Magnetic resonance T2* values of the myocardium are directly related to tissue iron levels. Minor effects from myocardial oxygenation and fibrosis are overwhelmed by the highly dominant iron effect in clinically relevant levels of myocardial iron overload. Myocardial T2* values less than 20 ms indicate iron overload, and this is considered severe when T2* is less than 10 ms. Decreasing myocardial T2* levels are associated with systolic and diastolic ventricular dysfunction. Most recorded cases of heart failure in thalassemia to date have occurred in patients with very low T2* values (in the severe range). Exceptions to this have occurred in patients with other causes of heart failure such as concomitant congenital heart disease. In patients presenting with heart failure who undergo aggressive chelation with continuous intravenous deferoxamine, longitudinal studies show that myocardial T2* increases, and this is accompanied by increases in ejection fraction and relief of heart failure. In cross-sectional studies, the myocardial T2* and ejection fraction of patients on deferiprone was superior to that of patients on deferoxamine. Randomized controlled prospective trials comparing these two drugs for their action in clearing myocardial iron, as measured by myocardial T2*, are under way and should report in 2005/2006. These trials will clarify the role of different chelators in the management of myocardial iron overload and may be valuable in reducing the toll of death in thalassemia from heart failure. PMID- 16339686 TI - Measurement and mapping of liver iron concentrations using magnetic resonance imaging. AB - Measurement of liver iron concentration (LIC) is an important clinical procedure in the management of transfusional iron overload with iron chelation. LIC gives an indication of over- or underchelation. Although chemical assay of needle biopsy samples from the liver has been considered the "gold standard" of LIC measurement, needle biopsy sampling errors can be surprisingly large owing to the natural spatial variation of LIC throughout the liver and the small size of biopsy specimens. A magnetic resonance imaging technique has now been developed that enables safe noninvasive measurement and imaging of LIC with a known accuracy and precision. Measurements of LIC can be made over the range of LIC encountered in clinical practice. The technique is based on the measurement and imaging of proton transverse relaxation rates (R2) within the liver. The R2 imaging technique can be implemented on most clinical 1.5-T MRI instruments, making it readily available to the clinical community. PMID- 16339689 TI - Rescued mice with Hb E transgene-developed red cell changes similar to human beta thalassemia/HbE disease. AB - A novel C57BL/6 transgenic murine model of HbE has been developed, and the heterotetrameric ((m)alpha2(h)beta(E)2) hemoglobin shows significant complementation of mild thalassemia phenotype in double heterozygous (beta(m+)beta(m-), beta(hE)) and homozygous knockout (beta(m-)beta(m-), beta(hE)) mice with 100% heterotetrameric hemoglobin. Lethal homozygous beta-thalassemic mice rescued by HbE transgenes mimic beta-thalassemia/HbE phenotype in human. Although anemia was not pronounced, other hematologic parameters were abnormally similar to beta-knockout mice. Flow cytometric study revealed a highly oxidative status in the red cells, but there were no marked changes in PS red cells and RBC vesicles. RBC life span and half-time of rescued red cells were shortened, indicating a rapid RBC destruction. PMID- 16339687 TI - Physiology and pathophysiology of iron cardiomyopathy in thalassemia. AB - Iron cardiomyopathy remains the leading cause of death in patients with thalassemia major. Magnetic resonance imaging (MRI) is ideally suited for monitoring thalassemia patients because it can detect cardiac and liver iron burdens as well as accurately measure left ventricular dimensions and function. However, patients with thalassemia have unique physiology that alters their normative data. In this article, we review the physiology and pathophysiology of thalassemic heart disease as well as the use of MRI to monitor it. Despite regular transfusions, thalassemia major patients have larger ventricular volumes, higher cardiac outputs, and lower total vascular resistances than published data for healthy control subjects; these hemodynamic findings are consistent with chronic anemia. Cardiac iron overload increases the relative risk of further dilation, arrhythmias, and decreased systolic function. However, many patients are asymptomatic despite heavy cardiac burdens. We explore possible mechanisms behind cardiac iron-function relationships and relate these mechanisms to clinical observations. PMID- 16339690 TI - Exploring the role of hepcidin, an antimicrobial and iron regulatory peptide, in increased iron absorption in beta-thalassemia. AB - To develop new treatments for beta-thalassemia, it is essential to identify the genes involved in the relevant pathophysiological processes. Iron metabolism in thalassemia mice being investigated, focusing on the expression of a gene called hepcidin (Hamp), which is expressed in the liver and whose product (Hamp) is secreted into the bloodstream. In mice, iron overload leads to overexpression of Hamp, while Hamp-knockout mice suffer from hemochromatosis. The aim of this study is to investigate Hamp in the mouse model of beta-thalassemia and to address the potential gene transfer of Hamp to prevent abnormal iron absorption. PMID- 16339691 TI - Murine and math models for the level of stable mixed chimerism to cure beta thalassemia by nonmyeloablative bone marrow transplantation. AB - Stable mixed chimeric stem cell transplantation in hemoglobinopathies exploits shorter erythroid survival in hemolytic anemias, providing normal donor red blood cells with a competitive survival advantage. This study examined the level of stable mixed chimerism necessary for complete hematological cure of the thalassemic phenotype, using a nonmyeloablative busulfan chemotherapeutic preparation. Thalassemic mice transplanted from congenic wild-type donors developed partial mixed chimerism. Hematologic cure required >80% donor red blood cells and only >13% donor white blood cells. Murine and human transplant results were compared with a math model for survival advantage of donor peripheral blood cells produced by steady-state chimeric marrow. PMID- 16339692 TI - Non-transferrin-bound iron during blood transfusion cycles in beta-thalassemia major. AB - Serum non-transferrin-bound iron (NTBI) levels assessed at arbitrary time points during transfusion cycles may not be representative if NTBI is undergoing significant changes during transfusion cycles. In 15 patients with beta thalassemia major (age: 21 +/- 6 years, liver iron concentration: 2200 +/- 1200 microg/g-liver), NTBI and other hematologic parameters (transferrin saturation, transferrin receptor) were measured weekly. The largest variation of NTBI levels between individual patients was observed at midcycle. For long-term monitoring of NTBI levels, a particular time point relative to the last blood transfusion should be selected for blood drawing. PMID- 16339693 TI - Genetic analysis of candidate modifier polymorphisms in Hb E-beta 0-thalassemia patients. AB - Hemoglobin E (Hb E)-beta-thalassemia patients display a range of clinical severities, from nearly asymptomatic to transfusion-dependent thalassemia major. Given this clinical heterogeneity, additional genetic factors modifying disease severity remain to be discovered. Association studies are being conducted to elucidate the role of genetic polymorphisms as disease severity modifiers in Hb E beta-thalassemia patients. Using strict scoring criteria, 1060 Hb E-beta thalassemia patients were categorized into mild, moderate, and severe groups. Taking a candidate gene approach, we found no statistically significant differences between the mild and severe patients groups in allelic or genotypic frequencies for single nucleotide polymorphisms within five genes known to influence globin gene expression and erythropoiesis. PMID- 16339694 TI - Measuring chromosome breaks in patients with thalassemia. AB - Iron-mediated oxidative stress plays an important role in the pathophysiology of thalassemia. Oxidative stress can cause lesions in DNA, including double-strand breaks. DNA damage, which is a cause of cancer (although not the only one), is recognized as deleterious. Unlike cancer, DNA damage can be assayed easily and relatively inexpensively in humans. In this study, a sensitive micronucleus assay was used to measure the frequency of chromosomal breaks in patients with alpha- and beta-thalassemia. The micronucleus test is based on the observation that a secondary nucleus (micronucleus) is formed around a chromosomal fragment, outside the main nucleus of a dividing cell. Micronuclei are readily apparent in red blood cells (RBCs), which otherwise lack DNA. We combined an immunomagnetic separation technique with single-laser flow cytometry to isolate and analyze reticulocytes in peripheral blood for the presence of micronuclei before these cells are removed by the spleen. Blood samples were obtained from patients with thalassemia and healthy volunteers. After immunomagnetic enrichment of CD71 positive reticulocytes, the cells were stained for micronuclei using the DNA dye 7-aminoactinomycin D (7-AAD) and evaluated by flow cytometry. Our findings indicate that higher levels of micronuclei frequencies are present in thalassemic RBCs. PMID- 16339695 TI - Longitudinal study of survival and causes of death in patients with thalassemia major in Greece. AB - Iron-induced organ degeneration is the main factor of mortality in patients with thalassemia major. Since chelation therapy is at a turning point, from the laborious parenteral route to the use of new promising oral agents, we investigated the current status of survival of these patients to present reliable data that will be useful in future comparative studies. Survival probabilities were estimated by the Kaplan-Meier method, and results were compared by the log rank test in a total of 647 thalassemic patients (pts) (52% males) born between 1/1/58 and 1/2/04. Terminal follow-up was 1/12/04. All transfusion-dependent pts monitored in our center, or in frequent contact if they had moved elsewhere, were strictly selected, excluding all rarely transfused or intermediate cases. Pts born before 1/1/75 were classified in group A (n = 366), while pts born later were included in group B (n = 281). According to the last 5 years' mean serum ferritin level, pts were divided into three hemosiderosis groups: (1) mild (<2000 microg/L) 49%, (2) moderate (2000-4000 microg/L) 28%, and (3) severe (>4000 microg/L) 23%. Of the 647 pts, 115 died (mean age: 22.6 +/- 6.2 years), most frequently by heart failure (71.3%) followed by sepsis (7.8%). Life expectancy in the entire population was up to 59% at 46 years. Survival was higher for pts born after 1975 than those before (P < .001). Statistically significantly different survival probabilities were found between groups with mild, moderate, or severe hemosiderosis (P < .001). Effective management with improved chelation therapy could lead to better results. PMID- 16339696 TI - Osteoporosis in beta-thalassemia: Clinical and genetic aspects. AB - Osteoporosis and osteopenia are frequent complications of thalassemia major (TM) and intermedia (TI). Osteoporosis was found in 23/25 patients with TI and in 115/239 patients with TM. In TM, no association was found with specific polymorphisms in candidate genes (vitamin D receptor, estrogen receptor, calcitonin receptor, and collagen type 1 alpha 1). Osteoporosis in female patients with TM was strongly associated with primary amenorrhea (P < .0001), while in male patients with TM, hypogonadism was not significantly related to bone mineral density (BMD) (P = .0001). Low BMD was also associated with cardiomiopathy (P = .01), diabetes mellitus (P = .0001), chronic hepatitis (P = .0029), and increased ALT (P = .01). PMID- 16339697 TI - Quality of life in patients with thalassemia intermedia compared to thalassemia major. AB - The impact of thalassemia major and thalassemia intermedia and their associated complications on quality of life (QOL) is largely unknown. Determining the degree of health impairment as perceived by the patient is essential information needed to recommend suitable therapy. The objective of this study was to evaluate QOL in transfusion-independent patients with thalassemia (non-Tx) compared with that in transfused patients (Tx) and to identify the factors that affect QOL in thalassemia. A convenient sample of 48 thalassemia patients (29 Tx and 19 non-Tx) with mean age of 14.6 years (SD = 7.5 years) were selected during a comprehensive visit to complete a Dartmouth Primary Care Cooperative Information Chart System (COOP) questionnaire. Patients rated QOL from excellent (1) to poor (5) on five dimensions of health status. Scores of 4 or 5 represent major limitations. These results were augmented by a brief medical history and chart review. Forty-one percent of Tx patients and 47% of non-Tx patients reported severe impairments in 1-6 and 1-2 domains, respectively. The most commonly reported affected domains were feelings such as anxiety, depression, and concern of overall health status or indications of recent deterioration in health. In contrast with previous beliefs, transfusion-independent thalassemia patients also suffer serious impairment in QOL. Presented data suggest that all patients with thalassemia undergo QOL assessment so that interventions focused on affected domains can be implemented. PMID- 16339698 TI - Low bone mineral density in adolescents with beta-thalassemia. AB - The pervasiveness of low bone mass (LBM) in beta-thalassemia (Thal) patients (pts) is escalating as the average life expectancy of these pts increases. Adolescence is a period of substantial bone accrual, which is crucial for future bone strength. Studies of LBM are prevalent among adults with Thal. However, limited information exists about bone accrual and LBM in adolescents with the disease. Thirty-one pts with beta-Thal (26 Thal major [TM], 5 Thal intermedia [TI]), aged 9-20 years (mean: 15.3 years), 14 males and 17 females, underwent measurement of spinal bone mineral density (BMD) by DEXA (Lunar, Prodigy). Height, weight, body mass index, and Tanner stage were assessed at the time of the BMD measurement. A total of 16.1% of the patients had normal bone mass (Z > or = -1), 22.6% had reduced bone mass (Z = -1 to -2), and 61.3% had low bone mass (Z < or = -2). BMD Z correlated with height and weight Z scores. Some 53.9% of subjects had normal gonadal function and 46.1% had induced puberty with gonadal steroids. BMD Z significantly worsened with age (P < .0001). However, there was no difference in the LBM prevalence between subjects with normal versus those with induced puberty: BMD Z was -2 or less in 71.4% of subjects with normal puberty versus 66.7% in those with induced puberty. Our results indicate a high prevalence of LBM among adolescents with Thal regardless of adequate transfusion and chelation regimens. Bone accrual was found to be suboptimal in adolescents with normal or induced puberty. Thus, calcium and vitamin D supplementation with antiresorptive therapies should be evaluated in the adolescent Thal pt with close monitoring of growth and sexual development. PMID- 16339699 TI - Does heterozygous beta-thalassemia confer a protection against coronary artery disease? AB - Six hundred and thirty-eight patients who presented with clinical symptoms and/or electrocardiographic findings suggestive of stable angina pectoris were studied; they were also investigated by coronary arteriography. Hemoglobin electrophoresis was performed on all patients to detect the presence of the beta-thalassemia trait. Results were analyzed by logistic regression analysis to determine whether the latter confers any protective effect against advanced coronary artery disease (aCAD; defined as the presence of atheromas in coronary arteries, resulting in stenosis at least 70%). The role of the currently accepted risk factors (smoking, hypertension, hypercholesterolemia, and diabetes) in developing aCAD were reconfirmed, while at the same time it was found that beta-thalassemia heterozygosity is associated with a reduced risk against aCAD (odds ratio 0.39, 95% confidence interval 0.16-0.98). The lipoprotein and blood rheology profile of these individuals may be the underlying causes of this protective effect. PMID- 16339700 TI - Therapeutic approaches to pulmonary hypertension in hemoglobinopathies: Efficacy and safety of sildenafil in the treatment of severe pulmonary hypertension in patients with hemoglobinopathy. AB - New approaches to the treatment of pulmonary arterial hypertension (PH) have increased symptomatic relief and prolonged survival. PH is a common sequela of the hemoglobinopathies, but the use of standard oral treatment options is limited because of toxicity and poor effectiveness. Sildenafil citrate is a selective and potent inhibitor of cGMP-specific phosphodiesterase-5 (PDE5), which promotes selective smooth muscle relaxation in lung vasculature and has been used successfully in the treatment of PH. Hemoglobinopathic patients suffering from severe PH who were treated with sildenafil citrate (50 mg b.i.d.) for periods ranging from 4 to 48 months showed a significant decrease in pulmonary pressure and improvement in exercise capacity and functional class. No significant adverse events were reported. These data, described in a small group of patients, indicate that sildenafil citrate is effective in the treatment of PH in hemoglobinopathies and is well tolerated long-term at a daily dose of 100 mg. PMID- 16339701 TI - Utility of Holter electrocardiogram in iron-overloaded hemoglobinopathies. AB - Cardiac arrhythmias are among the leading causes of morbidity and mortality of transfusion-dependent, iron-overloaded beta-thalassemia patients. Routine screening with Holter electrocardiogram has been recommended; however, infrequent electrocardiographic changes limit its clinical usefulness. The purpose of this study was to determine the diagnostic yield of Holter electrocardiogram monitoring and its correlation with patient symptoms and disease status. A retrospective analysis was performed on 27 transfusion-dependent thalassemia patients who underwent cardiac questionnaire and Holter screening yearly, in addition to echocardiogram and quantitative iron-level determination. Four patients had clinically significant arrhythmias detected on Holter screening, while 2 patients developed severe cardiac complications secondary to arrhythmias within 1 year of follow-up of normal Holter screening. Early detection of cardiac events among transfusion-dependent thalassemia patients with Holter electrocardiography is not clinically effective. Other screening modalities, including the transtelephonic event recorder, should be evaluated in arrhythmia surveillance. PMID- 16339702 TI - Hemolysis-associated pulmonary hypertension in thalassemia. AB - Accumulating evidence supports the existence of a condition involving hemolysis associated pulmonary hypertension (PHT). Hemolysis-induced release of cell-free hemoglobin and red blood cell arginase, resulting in impaired nitric oxide bioavailability, endothelial dysfunction, and PHT, has been reported in sickle cell disease. Since thalassemia is also a condition of chronic hemolysis, these patients are at risk. The data demonstrate that hemolysis-induced dysregulation of arginine metabolism and PHT also occurs in thalassemia. Erythrocyte release of arginase during hemolysis contributes to the development of PHT. Therapies that maximize arginine and nitric oxide bioavailability may benefit patients with thalassemia. PMID- 16339703 TI - A simple model to assess and improve adherence to iron chelation therapy with deferoxamine in patients with thalassemia. AB - Adherence to deferoxamine (DFO) is vital for the long-term survival of patients with thalassemia; however, currently no measure exists to quantify adherence directly. In this study, 90 patients with thalassemia major underwent liver iron concentration (LIC) assessment by SQUID biosusceptometer, were asked to rate their adherence to DFO using a Numerical Likert Scale (NLS), and were educated about complications of iron overload. Of 38% (n = 28) of patients who rated themselves as very compliant, 19 had elevated LIC related to inadequate dosing of DFO and nine reported nonadherence in the past. Adherence improved after counseling and LIC decreased by 25% (7-60%) in eight previously noncompliant patients who returned for subsequent LIC over 15 months. In conclusion, the NLS seems to be a simple but reliable tool to assess patients' adherence to DFO. Education and frequent noninvasive LIC assessments can improve adherence and iron burden. Elevated LIC does not necessarily reflect concurrent noncompliance; however, it can be an indication of nonadherence in the past. PMID- 16339704 TI - Preclinical and clinical development of deferitrin, a novel, orally available iron chelator. AB - Deferitrin, a novel, orally available iron chelator, is in the early stage of clinical development for the treatment of chronic iron overload due to transfusional therapy. Preclinical pharmacology studies demonstrate iron excretion largely by the fecal route. Initial clinical studies have shown deferitrin to be well absorbed. Further clinical studies are ongoing to determine the efficiency and safety of deferitrin. PMID- 16339705 TI - Allosensitization in patients receiving multiple blood transfusions. AB - Extended antigen (C, E, K) matching decreased the incidence of alloantibody (alloAB) and autoantibody (autoAB) formation, in addition to eliminating transfusion reactions in the multiply transfused sickle cell disease patients. AlloAB formation possibly transforms the immune system into a hyperactive state leading to further and earlier alloAB and autoAB formation. However, additional CEK matching results in marked overuse of Rh-negative packed red blood cell (pRBC) units, 30 minutes' extra time of a skilled technologist, and 153 dollars extra CEK reagent cost per unit to find CEK-matched pRBCs for every transfusion for these multiply transfused patients. PMID- 16339706 TI - Outcomes of preimplantation genetic diagnosis therapy in treatment of beta thalassemia: A retrospective analysis. AB - Thalassemia is one of the most common single-gene disorders that can be cured by hematopoietic stem cell transplantation (HCT) from a human leukocyte antigen (HLA)-identical sibling donor. In families that have an affected child, preimplantation genetic diagnosis (PGD) can be used to select an unaffected, HLA identical embryo. In brief, this procedure requires in vitro fertilization, oocyte retrieval, fertilization, and blastomere biopsy for identification of unaffected HLA-identical embryos. After delivery, umbilical cord blood from the sibling donor is collected for HCT. The objective of this study was to determine the outcomes of families using PGD therapy for cure of beta-thalassemia and to review the limitations of PGD therapy. Families affected with beta-thalassemia who attempted PGD therapy were retrospectively identified and reviewed for indication, attempted cycles, successful pregnancy, and transplantation outcomes. Eight identified families affected by thalassemia underwent PGD. The diagnosis of their affected children included six cases of beta-thalassemia major and two cases of transfusion-dependent hemoglobin E-beta-thalassemia patients. A total of 14 cycles of PGD were attempted, ranging from one to four attempts per family. Following successful identification of HLA-identical cells, two pregnancies occurred, of which one resulted in engraftment of a beta-thalassemia child. PGD therapy offers the possibility of recruiting a suitable donor for HCT, yet is limited by financial cost due to labor-intensive techniques, low probability of obtaining an HLA-matched unaffected embryo, variable implantation capacity, and significant emotional impact. Improvements in PGD therapy's efficacy and cost will make this a more viable option for affected families. PMID- 16339707 TI - Separation of normoblasts from whole blood by ultracentrifugation on arabino galactan discontinuous gradients: A tool for prenatal diagnosis. AB - Normoblasts can be separated at high resolution from whole blood by ultracentrifugation on discontinuous gradients of arabino-galactane after the red blood cells are removed by sedimentation. With this method, isolation of fetal normoblasts from maternal blood could provide a tool for rapid prenatal diagnosis. The availability of this noninvasive technique could avoid the cost and the risks of miscarriage that are associated with the current invasive procedures. PMID- 16339708 TI - Prenatal diagnosis of hemoglobinopathies in Ontario, Canada. AB - In 1989, the Province of Ontario established a molecular diagnostic laboratory for carrier detection and prenatal diagnosis of hemoglobinopathies. Over the past 15 years, the laboratory has provided prenatal diagnosis for 672 pregnancies at risk for severe hemoglobinopathies: 276 (41%) for homozygous beta-thalassemia or hemoglobin (Hb) E/beta-thalassemia, 211 (31%) for homozygous alpha 0-thalassemia (Hb Bart's hydrops fetalis), and/or Hb H disease, and 185 (28%) for various sickling disorders (Hb SS, Hb SC, Hb S/beta-thalassemia). Despite the availability of services for carrier screening, genetic counseling, and prenatal diagnosis, there has been only a modest reduction in the overall incidence of hemoglobinopathies in Ontario. PMID- 16339709 TI - The splenectomy controversy. PMID- 16339710 TI - Diagnosis and management of alpha thalassemia disorders. PMID- 16339711 TI - Management of the "difficult" iron-overloaded patient. PMID- 16339712 TI - Genotypes and phenotypes of thalassemia: A discussion. PMID- 16339713 TI - Nutrition: role in health and therapy. PMID- 16339714 TI - Oligomerization of G-protein-coupled receptors: lessons from the yeast Saccharomyces cerevisiae. PMID- 16339715 TI - Pde1 phosphodiesterase modulates cyclic AMP levels through a protein kinase A mediated negative feedback loop in Cryptococcus neoformans. AB - The virulence of the human pathogenic fungus Cryptococcus neoformans is regulated by a cyclic AMP (cAMP)-dependent protein kinase A (PKA) signaling cascade that promotes mating and the production of melanin and capsule. In this study, genes encoding homologs of the Saccharomyces cerevisiae low- and high-affinity phosphodiesterases, PDE1 and PDE2, respectively, were deleted in serotype A strains of C. neoformans. The resulting mutants exhibited moderately elevated levels of melanin and capsule production relative to the wild type. Epistasis experiments indicate that Pde1 functions downstream of the Galpha subunit Gpa1, which initiates cAMP-dependent signaling in response to an extracellular signal. Previous work has shown that the PKA catalytic subunit Pka1 governs cAMP levels via a negative feedback loop. Here we show that a pde1Delta pka1Delta mutant strain exhibits cAMP levels that are dramatically increased ( approximately 15 fold) relative to those in a pka1Delta single mutant strain and that a site directed mutation in a consensus PKA phosphorylation site reduces Pde1 function. These data provide evidence that fluctuations in cAMP levels are modulated by both Pka1-dependent regulation of Pde1 and another target that comprise a robust negative feedback loop to tightly constrain intracellular cAMP levels. PMID- 16339716 TI - A fungus-specific ras homolog contributes to the hyphal growth and virulence of Aspergillus fumigatus. AB - The Ras family of GTPase proteins has been shown to control morphogenesis in many organisms, including several species of pathogenic fungi. In a previous study, we identified a gene encoding a fungus-specific Ras subfamily homolog, rasB, in Aspergillus fumigatus. Here we report that deletion of A. fumigatus rasB caused decreased germination and growth rates on solid media but had no effect on total biomass accumulation after 24 h of growth in liquid culture. The DeltarasB mutant had an irregular hyphal morphology characterized by increased branching. Expression of rasBDelta113-135, a mutant transgene lacking the conserved rasB internal amino acid insertion, did not complement the deletion phenotype of delayed growth and germination rates and abnormal hyphal morphology. Virulence of the rasB deletion strain was diminished; mice infected with this strain exhibited approximately 65% survival compared to approximately 10% with wild-type and reconstituted strains. These data support the hypothesis that rasB homologs, which are highly conserved among fungi that undergo hyphal growth, control signaling modules important to the directional growth of fungal hyphae. PMID- 16339717 TI - Differential subcellular localization of members of the Toxoplasma gondii small heat shock protein family. AB - The results of this study describe the identification and characterization of the Toxoplasma gondii alpha-crystallin/small heat shock protein (sHsp) family. By database (www.toxodb.org) search, five parasite sHsps (Hsp20, Hsp21, Hsp28, Hsp29, and the previously characterized Hsp30/Bag1) were identified. As expected, they share the homologous alpha-crystallin domain, which is the key characteristic of sHsps. However, the N-terminal segment of each protein contains unique characteristics in size and sequence. Most T. gondii sHsps are constitutively expressed in tachyzoites and fully differentiated bradyzoites, with the exception of Hsp30/Bag1. Interestingly, by subcellular localization we observed that T. gondii sHsps are located in different compartments. Hsp20 is located at the apical end of the cell, Hsp28 is located inside the mitochondrion, Hsp29 showed a membrane-associated labeling, and Hsp21 appeared throughout the cytosol of the parasites. These particular differences in the immunostaining patterns suggest that their targets and functions might be different. PMID- 16339718 TI - Envoy, a PAS/LOV domain protein of Hypocrea jecorina (Anamorph Trichoderma reesei), modulates cellulase gene transcription in response to light. AB - Envoy, a PAS/LOV domain protein with similarity to the Neurospora light regulator Vivid, which has been cloned due to its lack of expression in a cellulase negative mutant, links cellulase induction by cellulose to light signaling in Hypocrea jecorina. Despite their similarity, env1 could not compensate for the lack of vvd function. Besides the effect of light on sporulation, we observed a reduced growth rate in constant light. An env1(PAS-) mutant of H. jecorina grows significantly slower in the presence of light but remains unaffected in darkness compared to the wild-type strain QM9414. env1 rapidly responds to a light pulse, with this response being different upon growth on glucose or glycerol, and it encodes a regulator essential for H. jecorina light tolerance. The induction of cellulase transcription in H. jecorina by cellulose is enhanced by light in the wild-type strain QM9414 compared to that in constant darkness, whereas a delayed induction in light and only a transient up-regulation in constant darkness of cbh1 was observed in the env1(PAS-) mutant. However, light does not lead to cellulase expression in the absence of an inducer. We conclude that Envoy connects the light response to carbon source signaling and thus that light must be considered an additional external factor influencing gene expression analysis in this fungus. PMID- 16339719 TI - Regulation and recovery of functions of Saccharomyces cerevisiae chaperone BiP/Kar2p after thermal insult. AB - We described earlier a novel mode of regulation of Hsp104, a cytosolic chaperone directly involved in the refolding of heat-denatured proteins, and designated it delayed upregulation, or DUR. When Saccharomyces cerevisiae cells grown at the physiological temperature of 24 degrees C, preconditioned at 37 degrees C, and treated briefly at 50 degrees C were shifted back to 24 degrees C, Hsp104 expression was strongly induced after 2.5 h of recovery and returned back to normal after 5 h. Here we show that the endoplasmic reticulum (ER) chaperones BiP/Kar2p and Lhs1p and the mitochondrial chaperone Hsp78 were also upregulated at the physiological temperature during recovery from thermal insult. The heat shock element (HSE) in the KAR2 promoter was found to be sufficient to drive DUR. The unfolded protein element could also evoke DUR, albeit weakly, in the absence of a functional HSE. BiP/Kar2p functions in ER translocation and assists protein folding. Here we found that the synthesis of new BiP/Kar2p molecules was negligible for more than an hour after the shift of the cells from 50 degrees C to 24 degrees C. Concomitantly, ER translocation was blocked, suggesting that preexisting BiP/Kar2p molecules or other necessary proteins were not functioning. Translocation resumed concomitantly with enhanced synthesis of BiP/Kar2p after 3 h of recovery, after which ER exit and protein secretion also resumed. For a unicellular organism like S. cerevisiae, conformational repair of denatured proteins is the sole survival strategy. Chaperones that refold proteins in the cytosol, ER, and mitochondria of S. cerevisiae appear to be subject to DUR to ensure survival after thermal insults. PMID- 16339720 TI - The polybasic juxtamembrane region of Sso1p is required for SNARE function in vivo. AB - Exocytosis in Saccharomyces cerevisiae requires the specific interaction between the plasma membrane t-SNARE complex (Sso1/2p;Sec9p)and a vesicular v-SNARE (Snc1/2p). While SNARE proteins drive membrane fusion, many aspects of SNARE assembly and regulation are ill defined. Plasma membrane syntaxin homologs (including Sso1p) contain a highly charged juxtamembrane region between the transmembrane helix and the "SNARE domain" or core complex domain. We examined this region in vitro and in vivo by targeted sequence modification, including insertions and replacements. These modified Sso1 proteins were expressed as the sole copy of Sso in S. cerevisiae and examined for viability. We found that mutant Sso1 proteins with insertions or duplications show limited function, whereas replacement of as few as three amino acids preceding the transmembrane domain resulted in a nonfunctional SNARE in vivo. Viability is also maintained when two proline residues are inserted in the juxtamembrane of Sso1p, suggesting that helical continuity between the transmembrane domain and the core coiled-coil domain is not absolutely required. Analysis of these mutations in vitro utilizing a reconstituted fusion assay illustrates that the mutant Sso1 proteins are only moderately impaired in fusion. These results suggest that the sequence of the juxtamembrane region of Sso1p is vital for function in vivo, independent of the ability of these proteins to direct membrane fusion. PMID- 16339722 TI - An Ustilago maydis septin is required for filamentous growth in culture and for full symptom development on maize. AB - During maize infection, the fungal pathogen Ustilago maydis undergoes a dimorphic transition from budding, yeast-like cells to a filamentous dikaryon that proliferates in the host. This transition is regulated by mating and environmental signals. Septation is likely to be important in the growth of the infectious dikaryon because of the need to maintain specific cellular compartments during dikaryotic growth. Recently, we found that the transcript level for a septin gene was influenced by the conserved cyclic AMP (cAMP)/protein kinase A signaling pathway that participates in regulating dimorphism in U. maydis. In this study, we describe the detailed analysis of the function of this septin gene, designated sep3, in the growth, development, and pathogenesis of U. maydis. We show that sep3 is required for normal cellular morphology and the division of budding haploid cells. The gene is also required for lipid-induced filamentous growth in culture but not during the formation of mating filaments on agar medium or in planta. Strains with a deletion in sep3 have a reduction in symptom development in maize, with filamentous cells in planta displaying morphological defects. In addition, sep3 influences the differentiation of hyphae into teliospores and the germination of these teliospores to produce the meiotic haploid progeny that complete the disease life cycle. Finally, the deletion of sep3 was found to influence the multiple budding phenotype of a mutant with a defect in the regulatory subunit of protein kinase A. This result is consistent with a link between sep3 and the control of morphogenesis by cAMP signaling. Overall, this study highlights the importance of regulating septation and changes in morphology during phytopathogenesis. PMID- 16339721 TI - Serial analysis of gene expression reveals conserved links between protein kinase A, ribosome biogenesis, and phosphate metabolism in Ustilago maydis. AB - The switch from budding to filamentous growth is a key aspect of invasive growth and virulence for the fungal phytopathogen Ustilago maydis. The cyclic AMP (cAMP) signaling pathway regulates dimorphism in U. maydis, as demonstrated by the phenotypes of mutants with defects in protein kinase A (PKA). Specifically, a mutant lacking the regulatory subunit of PKA encoded by the ubc1 gene displays a multiple-budded phenotype and fails to incite disease symptoms, although proliferation does occur in the plant host. A mutant with a defect in a catalytic subunit of PKA, encoded by adr1, has a constitutively filamentous phenotype and is nonpathogenic. We employed serial analysis of gene expression to examine the transcriptomes of a wild-type strain and the ubc1 and adr1 mutants to further define the role of PKA in U. maydis. The mutants displayed changes in the transcript levels for genes encoding ribosomal proteins, genes regulated by the b mating-type proteins, and genes for metabolic functions. Importantly, the ubc1 mutant displayed elevated transcript levels for genes involved in phosphate acquisition and storage, thus revealing a connection between cAMP and phosphate metabolism. Further experimentation indicated a phosphate storage defect and elevated acid phosphatase activity for the ubc1 mutant. Elevated phosphate levels in culture media also enhanced the filamentous growth of wild-type cells in response to lipids, a finding consistent with PKA regulation of morphogenesis in U. maydis. Overall, these findings extend our understanding of cAMP signaling in U. maydis and reveal a link between phosphate metabolism and morphogenesis. PMID- 16339723 TI - MYST family histone acetyltransferases in the protozoan parasite Toxoplasma gondii. AB - The restructuring of chromatin precedes tightly regulated events such as DNA transcription, replication, and repair. One type of chromatin remodeling involves the covalent modification of nucleosomes by histone acetyltransferase (HAT) complexes. The observation that apicidin exerts antiprotozoal activity by targeting a histone deacetyltransferase has prompted our search for more components of the histone modifying machinery in parasitic protozoa. We have previously identified GNAT family HATs in the opportunistic pathogen Toxoplasma gondii and now describe the first MYST (named for members MOZ, Ybf2/Sas3, Sas2, and Tip60) family HATs in apicomplexa (TgMYST-A and -B). The TgMYST-A genomic locus is singular and generates a approximately 3.5-kb transcript that can encode two proteins of 411 or 471 amino acids. TgMYST-B mRNA is approximately 7.0 kb and encodes a second MYST homologue. In addition to the canonical MYST HAT catalytic domain, both TgMYST-A and -B possess an atypical C2HC zinc finger and a chromodomain. Recombinant TgMYST-A exhibits a predilection to acetylate histone H4 in vitro at lysines 5, 8, 12, and 16. Antibody generated to TgMYST-A reveals that both the long and short (predominant) versions are present in the nucleus and are also plentiful in the cytoplasm. Moreover, both TgMYST-A forms are far more abundant in rapidly replicating parasites (tachyzoites) than encysted parasites (bradyzoites). A bioinformatics survey of the Toxoplasma genome reveals numerous homologues known to operate in native MYST complexes. The characterization of TgMYST HATs represents another important step toward understanding the regulation of gene expression in pathogenic protozoa and provides evolutionary insight into how these processes operate in eukaryotic cells in general. PMID- 16339724 TI - Gene set coregulated by the Saccharomyces cerevisiae nonsense-mediated mRNA decay pathway. AB - The nonsense-mediated mRNA decay (NMD) pathway has historically been thought of as an RNA surveillance system that degrades mRNAs with premature translation termination codons, but the NMD pathway of Saccharomyces cerevisiae has a second role regulating the decay of some wild-type mRNAs. In S. cerevisiae, a significant number of wild-type mRNAs are affected when NMD is inactivated. These mRNAs are either wild-type NMD substrates or mRNAs whose abundance increases as an indirect consequence of NMD. A current challenge is to sort the mRNAs that accumulate when NMD is inactivated into direct and indirect targets. We have developed a bioinformatics-based approach to address this challenge. Our approach involves using existing genomic and function databases to identify transcription factors whose mRNAs are elevated in NMD-deficient cells and the genes that they regulate. Using this strategy, we have investigated a coregulated set of genes. We have shown that NMD regulates accumulation of ADR1 and GAL4 mRNAs, which encode transcription activators, and that Adr1 is probably a transcription activator of ATS1. This regulation is physiologically significant because overexpression of ADR1 causes a respiratory defect that mimics the defect seen in strains with an inactive NMD pathway. This strategy is significant because it allows us to classify the genes regulated by NMD into functionally related sets, an important step toward understanding the role NMD plays in the normal functioning of yeast cells. PMID- 16339725 TI - Formation of an energized inner membrane in mitochondria with a gamma-deficient F1-ATPase. AB - Eukaryotic cells require mitochondrial compartments for viability. However, the budding yeast Saccharomyces cerevisiae is able to survive when mitochondrial DNA suffers substantial deletions or is completely absent, so long as a sufficient mitochondrial inner membrane potential is generated. In the absence of functional mitochondrial DNA, and consequently a functional electron transport chain and F(1)F(o)-ATPase, the essential electrical potential is maintained by the electrogenic exchange of ATP(4-) for ADP(3-) through the adenine nucleotide translocator. An essential aspect of this electrogenic process is the conversion of ATP(4-) to ADP(3-) in the mitochondrial matrix, and the nuclear-encoded subunits of F(1)-ATPase are hypothesized to be required for this process in vivo. Deletion of ATP3, the structural gene for the gamma subunit of the F(1)-ATPase, causes yeast to quantitatively lose mitochondrial DNA and grow extremely slowly, presumably by interfering with the generation of an energized inner membrane. A spontaneous suppressor of this slow-growth phenotype was found to convert a conserved glycine to serine in the beta subunit of F(1)-ATPase (atp2-227). This mutation allowed substantial ATP hydrolysis by the F(1)-ATPase even in the absence of the gamma subunit, enabling yeast to generate a twofold greater inner membrane potential in response to ATP compared to mitochondria isolated from yeast lacking the gamma subunit and containing wild-type beta subunits. Analysis of the suppressing mutation by blue native polyacrylamide gel electrophoresis also revealed that the alpha(3)beta(3) heterohexamer can form in the absence of the gamma subunit. PMID- 16339726 TI - Enzymes of the heme biosynthetic pathway in the nonphotosynthetic alga Polytomella sp. AB - Heme biosynthesis involves a number of enzymatic steps which in eukaryotes take place in different cell compartments. Enzyme compartmentalization differs between photosynthetic and nonphotosynthetic eukaryotes. Here we investigated the structures and subcellular localizations of three enzymes involved in the heme pathway in Polytomella sp., a colorless alga evolutionarily related to the green alga Chlamydomonas reinhardtii. Functional complementation of Escherichia coli mutant strains was used to isolate cDNAs encoding three heme biosynthetic enzymes, glutamate-1-semialdehyde aminotransferase, protoporphyrinogen IX oxidase, and ferrochelatase. All three proteins show highest similarity to their counterparts in photosynthetic organisms, including C. reinhardtii. All three proteins have N-terminal extensions suggestive of intracellular targeting, and immunoblot studies indicate their enrichment in a dense cell fraction that is enriched in amyloplasts. These results suggest that even though the plastids of Polytomella sp. are not photosynthetically active, they are the major site of heme biosynthesis. The presence of a gene for glutamate-1-semialdehyde aminotransferase suggests that Polytomella sp. uses the five-carbon pathway for synthesis of the heme precursor 5-aminolevulinic acid. PMID- 16339727 TI - Sequencing of random Euplotes crassus macronuclear genes supports a high frequency of +1 translational frameshifting. AB - Programmed translational frameshifts have been identified in genes from a broad range of organisms, but typically only a very few genes in a given organism require a frameshift for expression. In contrast, a recent analysis of gene sequences available in GenBank from ciliates in the genus Euplotes indicated that >5% required one or more +1 translational frameshifts to produce their predicted protein products. However, this sample of genes was nonrandom, biased, and derived from multiple Euplotes species. To test whether there truly is an abundance of frameshift genes in Euplotes, and to more accurately assess their frequency, we sequenced a random sample of 25 cloned genes/macronuclear DNA molecules from Euplotes crassus. Three new candidate +1 frameshift genes were identified in the sample that encode a membrane occupation and recognition nexus (MORN) repeat protein, a C(2)H(2)-type zinc finger protein, and a Ser/Thr protein kinase. Reverse transcription-PCR analyses indicate that all three genes are expressed in vegetatively proliferating cells and that the mRNAs retain the requirement of a frameshift. Although the sample of sequenced genes is relatively small, the results indicate that the frequency of genes requiring frameshifts in E. crassus is between 3.7% and 31.7% (at a 95% confidence interval). The current and past data also indicate that frameshift sites are found predominantly in genes that likely encode nonabundant proteins in the cell. PMID- 16339728 TI - RNA-binding domain proteins in Kinetoplastids: a comparative analysis. AB - RNA-binding proteins are important in many aspects of RNA processing, function, and destruction. One class of such proteins contains the RNA recognition motif (RRM), which consists of about 90 amino acid residues, including the canonical RNP1 octapeptide: (K/R)G(F/Y)(G/A)FVX(F/Y). We used a variety of homology searches to classify all of the RRM proteins of the three kinetoplastids Trypanosoma brucei, Trypanosoma cruzi, and Leishmania major. All three organisms have similar sets of RRM-containing protein orthologues, suggesting common posttranscriptional processing and regulatory pathways. Of the 75 RRM proteins identified in T. brucei, only 13 had clear homologues in other eukaryotes, although 8 more could be given putative functional assignments. A comparison with the 18 RRM proteins of the obligate intracellular parasite Encephalitozoon cuniculi revealed just 3 RRM proteins which appear to be conserved at the primary sequence level throughout eukaryotic evolution: poly(A) binding protein, the rRNA processing protein MRD1, and the nuclear cap binding protein. PMID- 16339729 TI - Transcriptome analysis of Paracoccidioides brasiliensis cells undergoing mycelium to-yeast transition. AB - Paracoccidioides brasiliensis is a thermodimorphic fungus associated with paracoccidioidomycosis (PCM), a systemic mycosis prevalent in South America. In humans, infection starts by inhalation of fungal propagules, which reach the pulmonary epithelium and transform into the yeast parasitic form. Thus, the mycelium-to-yeast transition is of particular interest because conversion to yeast is essential for infection. We have used a P. brasiliensis biochip carrying sequences of 4,692 genes from this fungus to monitor gene expression at several time points of the mycelium-to-yeast morphological shift (from 5 to 120 h). The results revealed a total of 2,583 genes that displayed statistically significant modulation in at least one experimental time point. Among the identified gene homologues, some encoded enzymes involved in amino acid catabolism, signal transduction, protein synthesis, cell wall metabolism, genome structure, oxidative stress response, growth control, and development. The expression pattern of 20 genes was independently verified by real-time reverse transcription PCR, revealing a high degree of correlation between the data obtained with the two methodologies. One gene, encoding 4-hydroxyl-phenyl pyruvate dioxygenase (4 HPPD), was highly overexpressed during the mycelium-to-yeast differentiation, and the use of NTBC [2-(2-nitro-4-trifluoromethylbenzoyl)-cyclohexane-1,3-dione], a specific inhibitor of 4-HPPD activity, as well as that of NTBC derivatives, was able to inhibit growth and differentiation of the pathogenic yeast phase of the fungus in vitro. These data set the stage for further studies involving NTBC and its derivatives as new chemotherapeutic agents against PCM and confirm the potential of array-based approaches to identify new targets for the development of alternative treatments against pathogenic microorganisms. PMID- 16339730 TI - Nd6p, a novel protein with RCC1-like domains involved in exocytosis in Paramecium tetraurelia. AB - In Paramecium tetraurelia, the regulated secretory pathway of dense core granules called trichocysts can be altered by mutation and genetically studied. Seventeen nondischarge (ND) genes controlling exocytosis have already been identified by a genetic approach. The site of action of the studied mutations is one of the three compartments, the cytosol, trichocyst, or plasma membrane. The only ND genes cloned to date correspond to mutants affected in the cytosol or in the trichocyst compartment. In this work, we investigated a representative of the third compartment, the plasma membrane, by cloning the ND6 gene. This gene encodes a 1,925-amino-acid protein containing two domains homologous to the regulator of chromosome condensation 1 (RCC1). In parallel, 10 new alleles of the ND6 gene were isolated. Nine of the 12 available mutations mapped in the RCC1-like domains, showing their importance for the Nd6 protein (Nd6p) function. The RCC1 protein is well known for its guanine exchange factor activity towards the small GTPase Ran but also for its involvement in membrane fusion during nuclear envelope assembly. Other proteins with RCC1-like domains are also involved in intracellular membrane fusion, but none has been described yet as involved in exocytosis. The case of Nd6p is thus the first report of such a protein with a documented role in exocytosis. PMID- 16339731 TI - Genetic and molecular analysis of phytochromes from the filamentous fungus Neurospora crassa. AB - Phytochromes (Phys) comprise a superfamily of red-/far-red-light-sensing proteins. Whereas higher-plant Phys that control numerous growth and developmental processes have been well described, the biochemical characteristics and functions of the microbial forms are largely unknown. Here, we describe analyses of the expression, regulation, and activities of two Phys in the filamentous fungus Neurospora crassa. In addition to containing the signature N terminal domain predicted to covalently associate with a bilin chromophore, PHY-1 and PHY-2 contain C-terminal histidine kinase and response regulator motifs, implying that they function as hybrid two-component sensor kinases activated by light. A bacterially expressed N-terminal fragment of PHY-2 covalently bound either biliverdin or phycocyanobilin in vitro, with the resulting holoprotein displaying red-/far-red-light photochromic absorption spectra and a photocycle in vitro. cDNA analysis of phy-1 and phy-2 revealed two splice isoforms for each gene. The levels of the phy transcripts are not regulated by light, but the abundance of the phy-1 mRNAs is under the control of the circadian clock. Phosphorylated and unphosphorylated forms of PHY-1 were detected; both species were found exclusively in the cytoplasm, with their relative abundances unaffected by light. Strains containing deletions of phy-1 and phy-2, either singly or in tandem, were not compromised in any known photoresponses in Neurospora, leaving their function(s) unclear. PMID- 16339732 TI - The plasmodial surface anion channel is functionally conserved in divergent malaria parasites. AB - The plasmodial surface anion channel (PSAC), a novel ion channel induced on human erythrocytes infected with Plasmodium falciparum, mediates increased permeability to nutrients and presumably supports intracellular parasite growth. Isotope flux studies indicate that other malaria parasites also increase the permeability of their host erythrocytes, but the precise mechanisms are unknown. Channels similar to PSAC or alternative mechanisms, such as the upregulation of endogenous host transporters, might fulfill parasite nutrient demands. Here we evaluated these possibilities with rhesus monkey erythrocytes infected with Plasmodium knowlesi, a parasite phylogenetically distant from P. falciparum. Tracer flux and osmotic fragility studies revealed dramatically increased permeabilities paralleling changes seen after P. falciparum infection. Patch-clamp of P. knowlesi-infected rhesus erythrocytes revealed an anion channel with striking similarities to PSAC: its conductance, voltage-dependent gating, pharmacology, selectivity, and copy number per infected cell were nearly identical. Our findings implicate a family of unusual anion channels highly conserved on erythrocytes infected with various malaria parasites. Together with PSAC's exposed location on the host cell surface and its central role in transport changes after infection, this conservation supports development of antimalarial drugs against the PSAC family. PMID- 16339733 TI - Identification and characterization of Cor33p, a novel protein implicated in tolerance towards oxidative stress in Candida albicans. AB - We applied two-dimensional gel electrophoresis to identify downstream effectors of CPH1 and EFG1 under hypha-inducing conditions in Candida albicans. Among the proteins that were expressed in wild-type cells but were strongly downregulated in a cph1Delta/efg1Delta double mutant in alpha-minimal essential medium at 37 degrees C, we could identify not-yet-characterized proteins, including Cor33-1p and Cor33-2p. The two proteins are almost identical (97% identity) and represent products of allelic isoforms of the same gene. Cor33p is highly similar to Cip1p from Candida sp. but lacks any significant homology to proteins from Saccharomyces cerevisiae. Strikingly, both proteins share homology with phenylcoumaran benzylic ether reductases and isoflavone reductases from plants. For other hypha-inducing media, like yeast-peptone-dextrose (YPD) plus serum at 37 degrees C, we could not detect any transcription for COR33 in wild-type cells, indicating that Cor33p is not hypha specific. In contrast, we found a strong induction for COR33 when cells were treated with 5 mM hydrogen peroxide. However, under oxidative conditions, transcription of COR33 was not dependent on EFG1, indicating that other regulatory factors are involved. In fact, upregulation depends on CAP1 at least, as transcript levels were clearly reduced in a Deltacap1 mutant strain under oxidative conditions. Unlike in wild-type cells, transcription of COR33 in a tsa1Delta mutant can be induced by treatment with 0.1 mM hydrogen peroxide. This suggests a functional link between COR33 and thiol specific antioxidant-like proteins that are important in the oxidative-stress response in yeasts. Concordantly, cor33Delta deletion mutants show retarded growth on YPD plates supplemented with hydrogen peroxide, indicating that COR33 in general is implicated in conferring tolerance toward oxidative stress on Candida albicans. PMID- 16339734 TI - Alternative sigma factors and their roles in bacterial virulence. AB - Sigma factors provide promoter recognition specificity to RNA polymerase holoenzyme, contribute to DNA strand separation, and then dissociate from the core enzyme following transcription initiation. As the regulon of a single sigma factor can be composed of hundreds of genes, sigma factors can provide effective mechanisms for simultaneously regulating expression of large numbers of prokaryotic genes. One newly emerging field is identification of the specific roles of alternative sigma factors in regulating expression of virulence genes and virulence-associated genes in bacterial pathogens. Virulence genes encode proteins whose functions are essential for the bacterium to effectively establish an infection in a host organism. In contrast, virulence-associated genes can contribute to bacterial survival in the environment and therefore may enhance the capacity of the bacterium to spread to new individuals or to survive passage through a host organism. As alternative sigma factors have been shown to regulate expression of both virulence and virulence-associated genes, these proteins can contribute both directly and indirectly to bacterial virulence. Sigma factors are classified into two structurally unrelated families, the sigma70 and the sigma54 families. The sigma70 family includes primary sigma factors (e.g., Bacillus subtilis sigma(A)) as well as related alternative sigma factors; sigma54 forms a distinct subfamily of sigma factors referred to as sigma(N) in almost all species for which these proteins have been characterized to date. We present several examples of alternative sigma factors that have been shown to contribute to virulence in at least one organism. For each sigma factor, when applicable, examples are drawn from multiple species. PMID- 16339735 TI - Molecules into cells: specifying spatial architecture. AB - A living cell is not an aggregate of molecules but an organized pattern, structured in space and in time. This article addresses some conceptual issues in the genesis of spatial architecture, including how molecules find their proper location in cell space, the origins of supramolecular order, the role of the genes, cell morphology, the continuity of cells, and the inheritance of order. The discussion is framed around a hierarchy of physiological processes that bridge the gap between nanometer-sized molecules and cells three to six orders of magnitude larger. Stepping stones include molecular self-organization, directional physiology, spatial markers, gradients, fields, and physical forces. The knowledge at hand leads to an unconventional interpretation of biological order. I have come to think of cells as self-organized systems composed of genetically specified elements plus heritable structures. The smallest self that can be fairly said to organize itself is the whole cell. If structure, form, and function are ever to be computed from data at a lower level, the starting point will be not the genome, but a spatially organized system of molecules. This conclusion invites us to reconsider our understanding of what genes do, what organisms are, and how living systems could have arisen on the early Earth. PMID- 16339736 TI - Ascospore formation in the yeast Saccharomyces cerevisiae. AB - Sporulation of the baker's yeast Saccharomyces cerevisiae is a response to nutrient depletion that allows a single diploid cell to give rise to four stress resistant haploid spores. The formation of these spores requires a coordinated reorganization of cellular architecture. The construction of the spores can be broadly divided into two phases. The first is the generation of new membrane compartments within the cell cytoplasm that ultimately give rise to the spore plasma membranes. Proper assembly and growth of these membranes require modification of aspects of the constitutive secretory pathway and cytoskeleton by sporulation-specific functions. In the second phase, each immature spore becomes surrounded by a multilaminar spore wall that provides resistance to environmental stresses. This review focuses on our current understanding of the cellular rearrangements and the genes required in each of these phases to give rise to a wild-type spore. PMID- 16339738 TI - Comparative genomic analyses of the bacterial phosphotransferase system. AB - We report analyses of 202 fully sequenced genomes for homologues of known protein constituents of the bacterial phosphoenolpyruvate-dependent phosphotransferase system (PTS). These included 174 bacterial, 19 archaeal, and 9 eukaryotic genomes. Homologues of PTS proteins were not identified in archaea or eukaryotes, showing that the horizontal transfer of genes encoding PTS proteins has not occurred between the three domains of life. Of the 174 bacterial genomes (136 bacterial species) analyzed, 30 diverse species have no PTS homologues, and 29 species have cytoplasmic PTS phosphoryl transfer protein homologues but lack recognizable PTS permeases. These soluble homologues presumably function in regulation. The remaining 77 species possess all PTS proteins required for the transport and phosphorylation of at least one sugar via the PTS. Up to 3.2% of the genes in a bacterium encode PTS proteins. These homologues were analyzed for family association, range of protein types, domain organization, and organismal distribution. Different strains of a single bacterial species often possess strikingly different complements of PTS proteins. Types of PTS protein domain fusions were analyzed, showing that certain types of domain fusions are common, while others are rare or prohibited. Select PTS proteins were analyzed from different phylogenetic standpoints, showing that PTS protein phylogeny often differs from organismal phylogeny. The results document the frequent gain and loss of PTS protein-encoding genes and suggest that the lateral transfer of these genes within the bacterial domain has played an important role in bacterial evolution. Our studies provide insight into the development of complex multicomponent enzyme systems and lead to predictions regarding the types of protein-protein interactions that promote efficient PTS-mediated phosphoryl transfer. PMID- 16339737 TI - Bacterial cell wall synthesis: new insights from localization studies. AB - In order to maintain shape and withstand intracellular pressure, most bacteria are surrounded by a cell wall that consists mainly of the cross-linked polymer peptidoglycan (PG). The importance of PG for the maintenance of bacterial cell shape is underscored by the fact that, for various bacteria, several mutations affecting PG synthesis are associated with cell shape defects. In recent years, the application of fluorescence microscopy to the field of PG synthesis has led to an enormous increase in data on the relationship between cell wall synthesis and bacterial cell shape. First, a novel staining method enabled the visualization of PG precursor incorporation in live cells. Second, penicillin binding proteins (PBPs), which mediate the final stages of PG synthesis, have been localized in various model organisms by means of immunofluorescence microscopy or green fluorescent protein fusions. In this review, we integrate the knowledge on the last stages of PG synthesis obtained in previous studies with the new data available on localization of PG synthesis and PBPs, in both rod shaped and coccoid cells. We discuss a model in which, at least for a subset of PBPs, the presence of substrate is a major factor in determining PBP localization. PMID- 16339741 TI - Physiological proteomics: cells, organs, biological fluids, and biomarkers. AB - Proteomic research is accelerating rapidly because of marked advances in protein labeling techniques, mass spectrometry (MS), and bioinformatics. Two-dimensional difference gel electrophoresis (2D-DIGE) is being used effectively in conjunction with liquid chromatography tandem MS (LC-MS/MS) and/or matrix-assisted laser desorption/ionization time-of-flight MS (MALDI-ToF MS) and database search software to quantify relative changes in the levels of proteins in two samples. It is now possible in a single study to identify and quantify large numbers of proteins and their posttranslational modifications in different biological samples. Comparisons can be made between groups of animals in different physiological states or in response to experimental treatment. Differences between normal individuals and those in disease states can form the foundation for elucidation of causative factors of disease and the identification of biomarkers for the diseased state. This symposium includes original research that compares the erythrocyte plasma membrane proteome in the normal and the sickle cell state, evaluates the anterior pituitary gland proteome in the ovariectomized rat in response to estrogen, and assesses proteomic methodology employed to identify potentially useful biomarkers in human cells and fluids for clinical medicine. It is directed not only to investigators working in these fields but also to a diverse group of scientists working in the biological and biomedical fields to stimulate cross-disciplinary awareness, interest, and collaboration. PMID- 16339742 TI - The proteomics of sickle cell disease: profiling of erythrocyte membrane proteins by 2D-DIGE and tandem mass spectrometry. AB - Quantitative changes in the red blood cell membrane proteome in sickle cell disease were analyzed using the two-dimensional fluorescence difference gel electrophoresis 2D-DIGE technique. From over 500 analyzed two-dimensional gel spots, we found 49 protein gel spots whose content in sickle cell membranes were changed by at least 2.5-fold as compared to control cells. In 38 cases we observed an increase and in 11 cases a decrease in content in the sickle cell membranes. The proteins of interest were identified by in-gel tryptic digestion followed by liquid chromatography in line with tandem mass spectrometry. From 38 analyzed gel spots, we identified 44 protein forms representing different modifications of 22 original protein sequences. The majority of the identified proteins fall into small groups of related proteins of the following five categories: actin accessory proteins--four proteins, components of lipid rafts- two proteins, scavengers of oxygen radicals--two proteins, protein repair participants--six proteins, and protein turnover components--three proteins. The number of proteins whose content in sickle RBC membrane is decreased is noticeably smaller, and most are either components of lipid rafts or actin accessory proteins. Elevated content of protein repair participants as well as oxygen radical scavengers may reflect the increased oxidative stress observed in sickle cells. PMID- 16339740 TI - The 2-hydroxycarboxylate transporter family: physiology, structure, and mechanism. AB - The 2-hydroxycarboxylate transporter family is a family of secondary transporters found exclusively in the bacterial kingdom. They function in the metabolism of the di- and tricarboxylates malate and citrate, mostly in fermentative pathways involving decarboxylation of malate or oxaloacetate. These pathways are found in the class Bacillales of the low-CG gram-positive bacteria and in the gamma subdivision of the Proteobacteria. The pathways have evolved into a remarkable diversity in terms of the combinations of enzymes and transporters that built the pathways and of energy conservation mechanisms. The transporter family includes H+ and Na+ symporters and precursor/product exchangers. The proteins consist of a bundle of 11 transmembrane helices formed from two homologous domains containing five transmembrane segments each, plus one additional segment at the N terminus. The two domains have opposite orientations in the membrane and contain a pore loop or reentrant loop structure between the fourth and fifth transmembrane segments. The two pore-loops enter the membrane from opposite sides and are believed to be part of the translocation site. The binding site is located asymmetrically in the membrane, close to the interface of membrane and cytoplasm. The binding site in the translocation pore is believed to be alternatively exposed to the internal and external media. The proposed structure of the 2HCT transporters is different from any known structure of a membrane protein and represents a new structural class of secondary transporters. PMID- 16339743 TI - Proteomics of the anterior pituitary gland as a model for studying the physiology of a heterogeneous organ. AB - The anterior pituitary gland (AP) secretes six established hormones that collectively control hundreds of biological and behavioral functions. Because of advances in mass spectrometry (MS), protein labeling, and bioinformatics, it is now possible to characterize, compare, and quantify the AP hormones together with large numbers of nonhormonal AP proteins. For example, by using high-performance liquid chromatography in line with tandem MS we characterized 145 proteins in sub cellular fractions of the AP of young adult male Golden Syrian hamsters and 115 proteins in subcellular fractions of the AP of young adult male mice. These included hormones, proteins involved in hormone synthesis and release, and housekeeping proteins. We also used difference gel electrophoresis in conjunction with MS and peptide mass fingerprinting to quantify the effects of estrogen on the AP-soluble protein fraction in rats. Ovariectomized rats were administered 50 microg of estradiol valerate subcutaneously and studied 48 hrs later, before the onset of the anticipated surges of gonadotropins in blood. Following DeCyder image analysis, we identified by MS and peptide mass fingerprinting 26 protein spots that were upregulated and 19 protein spots that were downregulated. Estrogen increased levels of acidic isoforms of growth hormone and prolactin, several proteins involved in protein synthesis, folding and secretion, and several metabolic enzymes. Most of the downregulated proteins are involved in RNA or DNA interactions. We followed up on the results with RT-PCR and immunohistochemical techniques to demonstrate that one protein identified by MS in hamster AP, fertility protein SP22, is synthesized in the AP and localized primarily in somatotropes and thyrotropes. These experiments demonstrate the efficacy of our proteomics approach to characterize AP proteins and quantify changes in them. The approaches used to study the AP could serve as a model to investigate other heterogeneous organs. PMID- 16339744 TI - Estrogen regulation of the rat anterior pituitary gland proteome. AB - Estrogen is known to affect the regulation of all six of the established anterior pituitary gland (AP) hormones, but little is known of the specifics of its regulation of the AP hormones, their isoforms, and nonhormonal AP proteins. We used difference gel electrophoresis in conjunction with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and peptide mass fingerprinting to quantify the effects of estrogen on the AP-soluble protein fraction in rats. Two-month-old rats were ovariectomized and used at 6 months of age. They were injected subcutaneously with sesame oil vehicle or 50 mug estradiol valerate in vehicle and studied 48 hrs later, approximately 3 hrs before the time of the anticipated onset of the estrogen-induced surges of gonadotropins in blood. The APs were pooled, and the soluble protein fraction was examined in replicate analyses. After DeCyder software analysis, we identified 26 protein spots that had a 1.5-fold or greater average increase in the experimental group relative to the controls. Nineteen showed a 1.5-fold or greater decrease. Estrogen increased levels of the more acidic isoforms of growth hormone and prolactin and of proteins involved in protein synthesis, folding, and secretion (e.g., eukaryotic translation elongation factor 2, ERp57, ERp29, Hsc70-ps1, calreticulin, coatomer delta subunit, and secretogranin II) and of some metabolic enzymes (e.g., arginosuccinate synthetase, enolase 1, creatine kinase B, phosphoglycerate mutase, malate dehydrogenase, pyruvate kinase, and aldolase A). The majority of the downregulated proteins were involved in RNA or DNA interactions (e.g., five heterogeneous nuclear ribonucleoproteins, DEAD-box proteins 17 and 48, ssDNA binding protein PUR-alpha, PTB-associated splicing factor, and Pigpen protein), but isovaleryl coenzyme A dehydrogenase, mitochondrial aldehyde dehydrogenase, stathmin 1, vinculin, radixin, and secretogranin III were also reduced. Our results indicate that estrogen acts in vivo within 48 hrs to modulate levels of a significant number of AP proteins. PMID- 16339745 TI - Proteomics as a tool for clinically relevant biomarker discovery and validation. AB - The excitement associated with clinical applications of proteomics was initially focused on its potential to serve as a vehicle for both biomarker discovery and drug discovery and routine clinical sample analysis. Some approaches were thought to be able to "identify" mass spectral characteristics that distinguished between control and disease samples, and thereafter it was believed that the same tool could be employed to screen samples in a high-throughput clinical setting. However, this has been difficult to achieve, and the early promise is yet to be fully realized. While we see an important place for mass spectrometry in drug and biomarker discovery, we believe that alternative strategies will prove more fruitful for routine analysis. Here we discuss the power and versatility of 2D gels and mass spectrometry in the discovery phase of biomarker work but argue that it is better to rely on immunochemical methods for high-throughput validation and routine assay applications. PMID- 16339746 TI - Ovarian aging and menopause: current theories, hypotheses, and research models. AB - Aging of the reproductive system has been studied in numerous vertebrate species. Although there are wide variations in reproductive strategies and hormone cycle components, many of the fundamental changes that occur during aging are similar. Evolutionary hypotheses attempt to explain why menopause occurs, whereas cellular hypotheses attempt to explain how it occurs. It is commonly believed that a disruption in the hypothalamic-pituitary-gonadal axis is responsible for the onset of menopause. Data exist to demonstrate that the first signs of menopause occur at the level of the brain or the ovary. Thus, finding an appropriate and representative animal model is especially important for the advancement of menopause research. In primates, there is a gradual decline in the function of the hypothalamic-pituitary-gonadal (HPG) axis ultimately resulting in irregularities in menstrual cycles and increasingly sporadic incidence of ovulation. Rodents also exhibit a progressive deterioration in HPG axis function; however, they also experience a period of constant estrus accompanied by intermittent ovulations, reduced progesterone levels, and elevated circulating estradiol levels. It is remarkable to observe that females of other classes also demonstrate deterioration in HPG axis function and ovarian failure. Comparisons of aging in various taxa provide insight into fundamental biological mechanisms of aging that could underlie reproductive decline. PMID- 16339747 TI - Molecular aspects of arterial smooth muscle contraction: focus on Rho. AB - The vascular smooth muscle cell is a highly specialized cell whose primary function is contraction and relaxation. It expresses a variety of contractile proteins, ion channels, and signalling molecules that regulate contraction. Upon contraction, vascular smooth muscle cells shorten, thereby decreasing the diameter of a blood vessel to regulate the blood flow and pressure. Contractile activity in vascular smooth muscle cells is initiated by a Ca(2+)-calmodulin interaction to stimulate phosphorylation of the light chain of myosin. Ca(2+) sensitization of the contractile proteins is signaled by the RhoA/Rho-kinase pathway to inhibit the dephosphorylation of the light chain by myosin phosphatase, thereby maintaining force. Removal of Ca(2+) from the cytosol and stimulation of myoson phosphatase initiate the relaxation of vascular smooth muscle. PMID- 16339739 TI - Coronavirus pathogenesis and the emerging pathogen severe acute respiratory syndrome coronavirus. AB - Coronaviruses are a family of enveloped, single-stranded, positive-strand RNA viruses classified within the Nidovirales order. This coronavirus family consists of pathogens of many animal species and of humans, including the recently isolated severe acute respiratory syndrome coronavirus (SARS-CoV). This review is divided into two main parts; the first concerns the animal coronaviruses and their pathogenesis, with an emphasis on the functions of individual viral genes, and the second discusses the newly described human emerging pathogen, SARS-CoV. The coronavirus part covers (i) a description of a group of coronaviruses and the diseases they cause, including the prototype coronavirus, murine hepatitis virus, which is one of the recognized animal models for multiple sclerosis, as well as viruses of veterinary importance that infect the pig, chicken, and cat and a summary of the human viruses; (ii) a short summary of the replication cycle of coronaviruses in cell culture; (iii) the development and application of reverse genetics systems; and (iv) the roles of individual coronavirus proteins in replication and pathogenesis. The SARS-CoV part covers the pathogenesis of SARS, the developing animal models for infection, and the progress in vaccine development and antiviral therapies. The data gathered on the animal coronaviruses continue to be helpful in understanding SARS-CoV. PMID- 16339748 TI - Long-term prolactin exposure differentially stimulated the transcellular and solvent drag-induced calcium transport in the duodenum of ovariectomized rats. AB - Prolactin, having been shown to stimulate transcellular active and solvent drag induced calcium transport in the duodenum of female rats, was postulated to improve duodenal calcium transport in estrogen-deficient rats. The aim of the present study was, therefore, to demonstrate the effects of long-term prolactin exposure produced by anterior pituitary (AP) transplantation on the duodenal calcium transport in young (9-week-old) and adult (22-week-old) ovariectomized rats. We found that ovariectomy did not alter the transcellular active duodenal calcium transport in young and adult rats fed normal calcium diet (1.0% w/w Ca) but decreased the solvent drag-induced duodenal calcium transport from 75.50 +/- 10.12 to 55.75 +/- 4.77 nmol.hr(-1).cm(-2) (P < 0.05) only in adult rats. Long term prolactin exposure stimulated the transcellular active calcium transport in young and adult AP-grafted ovariectomized rats fed with normal calcium diet by more than 2-fold from 7.56 +/- 0.79 to 16.54 +/- 2.05 (P < 0.001) and 9.78 +/- 0.72 to 15.99 +/- 1.75 (P < 0.001) nmol.hr(-1).cm(-2), respectively. However, only the solvent drag-induced duodenal calcium transport in young rats was enhanced by prolactin from 95.51 +/- 10.64 to 163.20 +/- 18.03 nmol.hr(-1).cm(-2) (P < 0.001) whereas that in adult rats still showed a decreased flux from 75.50 +/- 10.12 to 47.77 +/- 5.42 nmol.hr(-1).cm(-2) (P < 0.05). Because oral calcium supplement has been widely used to improve calcium balance in estrogen-deficient animals, the effect of a high-calcium diet (2.0% w/w Ca) was also investigated. The results showed that stimulatory action of long-term prolactin on the transcellular active duodenal calcium transport in both young and adult rats was diminished after being fed a high-calcium diet. The same diet also abolished prolactin-enhanced solvent drag-induced duodenal calcium transport in young and further decreased that in adult AP-grafted ovariectomized rats. We concluded that the solvent drag-induced duodenal calcium transport in adult rats was decreased after ovariectomy. Long-term prolactin exposure stimulated the transcellular active duodenal calcium transport in both young and adult rats whereas enhancing the solvent drag-induced duodenal calcium transport only in young rats. Effects of prolactin were abolished by a high-calcium diet. PMID- 16339750 TI - Gender modulates the expression of calcium-regulating proteins in pediatric atrial myocardium. AB - A differential expression of sarcoplasmic reticulum calcium-ATPase (SERCA2a) and phospholamban (PLB) characterizes the remodeling process in heart failure and atrial arrhythmias in adult patients. Gender is known to modulate the course and prognosis of different forms of heart disease. We hypothesized that gender plays a role in molecular changes of myocardial calcium regulating components already in childhood. Moreover, we studied the influence of volume overloaded (VO) on SERCA2a and PLB in pediatric patients. Quantitative reverse transcription polymerase chain reaction was used to measure mRNA expression of SERCA2a and PLB in atrial myocardium from 30 pediatric patients (12 girls, 18 boys). Eighteen patients had VO right atria, and 12 patients had not-overloaded atria (NO). Protein expression was studied by Western blot. In the entire population, SERCA2a and PLB expression was not different between girls and boys. If hemodynamic overload was taken into account, SERCA2a mRNA expression was significantly reduced in the VO group compared with the NO group (P = 0.021). The VO versus NO difference was restricted to boys, which corresponds to a highly significant interaction of gender versus VO status (P = 0.002). The PLB to SERCA2a protein ratio was significantly lower in girls (P = 0.028). The decrease in SERCA2a mRNA expression in VO atrial myocardium and the PLB to SERCA2a ratio of protein expression was modulated by gender in this pediatric population. To our knowledge, this study is the first to show the impact of gender on the differential expression of calcium-regulating components in pediatric cardiac patients. PMID- 16339749 TI - Possibility of autocrine beta-adrenergic signaling in C2C12 myotubes. AB - Levodopa reportedly inhibits insulin action in skeletal muscle. Here we show that C2C12 myotubes produce levodopa and that insulin-stimulated glucose transport is enhanced when endogenous levodopa is depleted. Exogenous levodopa prevented the stimulation of glucose transport by insulin (P < 0.05) and increased cAMP concentrations (P < 0.05). The decrease in insulin-stimulated glucose transport caused by levodopa was attenuated by propranolol (a beta-adrenergic antagonist) and prevented by NSD-1015 (NSD), an inhibitor of DOPA decarboxylase (DDC; converts levodopa to dopamine). Propranolol and NSD both prevented levodopa related increases in [cAMP]. However, the effects of levodopa were unlikely to be dependent on the conversion of levodopa to catecholamines because we could detect neither DDC in myotubes nor catecholamines in media after incubation of myotubes with levodopa. The data suggest the possibility of novel autocrine beta adrenergic action in C2C12 myotubes in which levodopa, produced by myotubes, could have hormone-like effects that impinge on glucose metabolism. PMID- 16339751 TI - Tissue distribution of fetal liver cells following in utero transplantation in mice. AB - Transplantation of hepatic stem cells in utero has been advanced as a potential clinical approach to a variety of diseases, including deficiencies of coagulation factors. Although syngeneic transplantation has met with some success, consideration needs to be given to the potential for transplanted cells to colonize nontarget tissues. Liver cells were harvested from Rosa26 embyros at embryonic age 12.5 days postconception (pc) and transplanted into the peritoneal cavity of syngeneic recipients in utero. Tissues were harvested from tissue recipients at various time points ranging from 1 to 328 days pc, and tissues were stained for beta-galactosidase to identify the existence of cells derived from Rosa26 donors. Beta-galactosidase-positive cells were found in the lung, liver, and brain as early as 20 days pc and through 328 days pc. Positive cells in these tissues existed as islands of cells that were morphologically similar to hepatocytes. In the spleen, individual beta-galactosidase-positive cells of both leukocytic and erythrocytic lineages were present, and suggest that hematopoietic cells were transferred to recipients along with hepatocytes. The lack of an inflammatory response to the beta-galactosidase-positive cells suggests that the donor cells were immunologically tolerated. In summary, the possibility that cells administered in utero may inadvertently colonize nontarget tissues suggests that clinical application of this method will need to be approached with diligence. PMID- 16339752 TI - Distinct host-dependent pathogenic mechanisms leading to a similar clinical anemia after infection with lymphocytic choriomeningitis virus. AB - The Docile strain of lymphocytic choriomeningitis virus (LCMV) induces anemia in a number of inbred strains of mice, including C3HeB/FeJ and CBA/Ht animals. A difference in the kinetics of anemia and in compensatory reticulocytosis suggested that impaired erythropoiesis was the major pathogenic mechanism involved in CBA/Ht mice, but not in C3HeB/FeJ mice. In both mouse strains an antierythrocyte autoantibody production that depended on the presence of functional CD4+ T lymphocytes was observed. Although depletion of T helper lymphocytes prevented anemia in C3HeB/FeJ mice, this treatment largely failed to inhibit the development of the disease in CBA/Ht animals. This observation indicated that the antierythrocyte autoimmune response induced by the infection was at least partly responsible for the anemia of C3HeB/FeJ mice, but not of CBA/Ht mice. Erythrophagocytosis was enhanced in both mouse strains after LCMV infection, but did not appear to be a major cause of anemia. These data clearly indicate that similar disease profiles induced by the same virus in two different host strains can be the result of distinctly different mechanisms. PMID- 16339753 TI - The HIV-1 Tat protein enhances megakaryocytic commitment of K562 cells by facilitating CREB transcription factor coactivation by CBP. AB - The human immunodeficiency virus type 1 (HIV-1) Tat protein regulates transcription factor functions and alters cellular gene expression. Because hematopoietic progenitor cell (HPC) differentiation requires activation of lineage-specific transcription factors, Tat may affect hematopoiesis in HIV-1 infected micro-environments. We have monitored the molecular effects of Tat on megakaryocytic differentiation in the HPC line, K562. Flow cytometry analysis of CD61 indicated that phorbol myristate acetate (PMA) (16 nM) stimulated megakaryocytic commitment of K562 cells was increased (3- to 4-fold) following exposure to Tat (1-100 ng/ml). Activation of the megakaryocytic transcription factor cAMP regulatory element binding protein (CREB) and its coactivation by the CREB binding protein (CBP) was subsequently monitored. CREB phosphorylation and DNA binding were measured by Western immunodetection and electrophoretic mobility shift assays (EMSA), respectively. Within 2 hrs after stimulation, Tat increased both CREB phosphorylation and DNA binding by 7- to 10-fold. Transient cotransfection with CREB reporter and CBP expression plasmids demonstrated that Tat treatment increases (3- to 4-fold) both PMA-stimulated and CBP-mediated transcription via the cAMP regulatory element. Histone acetyl transferase (HAT) activity was increased (8- to 10-fold) in Tat-stimulated cells, which suggested increased chromosomal accessibility of transcription factors. Two-hybrid cotransfection assays using reporter plasmid containing the GAL4 DNA-binding domain and expression plasmid coding for the GAL4-CBP fusion protein, showed that Tat increases (2-fold) CBP-mediated coactivation of CREB. Both reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis showed that Tat treatment increases CBP gene expression (7- to 9-fold) and protein levels (5- to 7-fold) within 6-12 hrs after stimulation. Our findings indicated that Tat treatment increases both CREB function and CREB coactivation by CBP, which may facilitate megakaryocytic commitment of K562 cells. Induction of this molecular signaling by HIV-1 Tat protein may have relevance in understanding the HIV-induced hematologic manifestations and possibly in regulation of viral infectivity parameters in progenitor cell reservoirs. PMID- 16339754 TI - Global role of the immune system in identifying cancer initiation and limiting disease progression. PMID- 16339755 TI - Fine needle aspiration is a feasible and accurate technique in the diagnosis of lymphoma. PMID- 16339756 TI - TP53 mutation and microsatellite instability status for the prediction of survival in adjuvant-treated colon cancer patients. PMID- 16339757 TI - Mass spectrometric approach for screening modifications of total serum N-glycome in human diseases: application to cirrhosis. AB - Congenital and acquired modifications of glycosylation in diseases are a rapidly growing field that demonstrates the importance of glycosylation in human biology. Unfortunately, in clinical biochemistry, very few tests are available to explore oligosaccharide metabolism on a large scale. Such an assay needs to be of high throughput, rapid, and preferentially noninvasive. In the present study, we describe a method to analyze qualitative variations of N-glycosylation of human serum proteins. The method is based on direct release of N-linked oligosaccharides from patient serum samples, a single-step purification, and a matrix-assisted laser desorption ionization time of flight mass spectrometric analysis. A complementary structural study of the released oligosaccharides was achieved by enzymatic digestions, linkage analysis, and electrospray ionization ion trap mass spectrometry (ESI-IT-MS) of the permethylated N-glycome. A total of 26 oligosaccharide structures were individualized, their presence in human serum being the result of the combination of the biosynthesis and catabolic pathways. Application of the protocol to the serum of patients with cirrhosis demonstrates the ability of this assay to identify acquired modifications of glycosylation. Furthermore, we have analyzed the N-glycans and showed the increase in bisecting N-acetylglucosamine residue, core fucosylation, and the presence of an important population of neutral oligosaccharides. The study of total serum N-glycome modifications is a preliminary for the discovery of new noninvasive diagnostic or prognostic biomarkers resulting from the variations of the N-glycan metabolism during diseases. PMID- 16339758 TI - The differential engagement of arrestin surface charges by the various functional forms of the receptor. AB - G-protein-coupled receptor signaling is terminated by arrestin proteins that preferentially bind to the activated phosphorylated form of the receptor. Arrestins also bind active unphosphorylated and inactive phosphorylated receptors. Binding to the non-preferred forms of the receptor is important for visual arrestin translocation in rod photoreceptors and the regulation of receptor signaling and trafficking by non-visual arrestins. Given the importance of arrestin interactions with the various functional forms of the receptor, we performed an extensive analysis of the receptor-binding surface of arrestin using site-directed mutagenesis. The data indicated that a large number of surface charges are important for arrestin interaction with all forms of the receptor. Arrestin elements involved in receptor binding are differentially engaged by the various functional forms of the receptor, each requiring a unique subset of arrestin residues in a specific spatial configuration. We identified several additional phosphate-binding elements in the N-domain and demonstrated for the first time that the active receptor preferentially engages the arrestin C-domain. We also found that the interdomain contact surface is important for arrestin interaction with the non-preferred forms of the receptor and that residues in this region play a role in arrestin transition into its high affinity receptor binding state. PMID- 16339759 TI - Activation of the RNA-dependent protein kinase PKR promoter in the absence of interferon is dependent upon Sp proteins. AB - The protein kinase regulated by RNA (PKR) is interferon (IFN)-inducible and plays important roles in many cellular processes, including virus multiplication, cell growth, and apoptosis. The TATA-less PKR promoter possesses a novel 15-bp DNA element (kinase conserved sequence (KCS)) unique to the human and mouse PKR genes that is conserved in sequence and position. We found that Sp1 and Sp3 of the Sp family of transcription factors bind at the KCS element. Their involvement was analyzed in the activation of basal and IFN-inducible PKR promoter activity. Both the small and large isoforms of Sp3 co-purified with KCS protein binding activity (KBP) by using nuclear extracts from HeLa cells not treated with IFN. Two forms of the KCS-binding protein complex were demonstrated by electrophoretic mobility shift assay analysis; one contained Sp1 and the other Sp3. In mouse cells null for all Sp3 isoforms, PKR expression was reduced to approximately 50% that of wild-type cells in the absence of IFN. The IFN-inducible expression of PKR, however, was Sp3-independent but STAT1- and JAK1-dependent. Overexpression of Sp1 in human U cells resulted in increased PKR promoter activity. In Drosophila SL2 cells lacking Sp proteins, both Sp1 and Sp3 large but not small isoforms activated PKR promoter expression, with the Sp1-mediated activation dominant. Mutational analysis of the PKR promoter region indicated a cooperative interaction between two different Sp sites, one of which is within the KCS element. These results establish that, in the absence of IFN treatment, activation of PKR basal expression is mediated by Sp1 and Sp3 proteins in a cooperative manner. PMID- 16339760 TI - Interaction of Huntingtin-associated protein-1 with kinesin light chain: implications in intracellular trafficking in neurons. AB - Huntingtin-associated protein-1 (HAP1) was initially identified as an interacting partner of huntingtin, the Huntington disease protein. Unlike huntingtin that is ubiquitously expressed throughout the brain and body, HAP1 is enriched in neurons, suggesting that its dysfunction could contribute to Huntington disease neuropathology. Growing evidence has demonstrated that HAP1 and huntingtin are anterogradely transported in axons and that the abnormal interaction between mutant huntingtin and HAP1 may impair axonal transport. However, the exact role of HAP1 in anterograde transport remains unclear. Here we report that HAP1 interacts with kinesin light chain, a subunit of the kinesin motor complex that drives anterograde transport along microtubules in neuronal processes. The interaction of HAP1 with kinesin light chain is demonstrated via a yeast two hybrid assay, glutathione S-transferase pull down, and coimmunoprecipitation. Furthermore, HAP1 is colocalized with kinesin in growth cones of neuronal cells. We also demonstrated that knocking down HAP1 via small interfering RNA suppresses neurite outgrowth of PC12 cells. Analysis of live neuronal cells with fluorescence microscopy and fluorescence recovery after photobleaching demonstrates that suppressing the expression of HAP1 or deleting the HAP1 gene inhibits the kinesin-dependent transport of amyloid precursor protein vesicles. These studies provide a molecular basis for the participation of HAP1 in anterograde transport in neuronal cells. PMID- 16339761 TI - New insights into the evolutionary links relating to the 3-deoxy-D-arabino heptulosonate 7-phosphate synthase subfamilies. AB - Bacterial 3-deoxy-d-arabino-heptulosonate 7-phosphate synthases (DAHPSs) have been divided into either of two classes (Class I/Class II) or subfamilies (AroAI(alpha)/AroAI(beta)). Our investigation into the biochemical properties of the unique bifunctional DAHPS from Bacillus subtilis provides new insight into the evolutionary link among DAHPS subfamilies. In the present study, the DAHPS (aroA) and chorismate mutase (aroQ) activities of B. subtilis DAHPS are separated by domain truncation. Detailed enzymatic studies with the full-length wild-type protein and the truncated domains led to our hypothesis that the aroQ domain was fused to the N terminus of aroA in B. subtilis during evolution for the purpose of feedback regulation and not for the creation of a bona fide bifunctional enzyme. In addition, examination of aroA and aroQ fusion proteins from Porphyromonas gingivalis, in which the aroQ domain is fused to the C terminus of aroA, further supports the hypothesis. These results, along with sequence structure analysis of the DAHPS families suggest that "feedback regulation" may indeed be the evolutionary link between the two classes/subfamilies. It is likely that DAHPSs evolved from a primitive unregulated member of the AroAI(beta) subfamily. During evolution, some members of the AroAI(beta) subfamily remained unregulated, whereas other members acquired an extra domain for feedback regulation. The AroAI(alpha) subfamilies, however, evolved in a more complex manner to acquire insertions/extensions in the (beta/alpha)(8) barrel to function as regulatory elements. PMID- 16339762 TI - Glutamine binding opens the ammonia channel and activates glucosamine-6P synthase. AB - Glucosamine-6P synthase catalyzes the synthesis of glucosamine-6P from fructose 6P and glutamine and uses a channel to transfer ammonia from its glutaminase to its synthase active site. X-ray structures of glucosamine-6P synthase have been determined at 2.05 Angstroms resolution in the presence of fructose-6P and at 2.35 Angstroms resolution in the presence of fructose-6P and 6-diazo-5-oxo-L norleucine, a glutamine affinity analog that covalently modifies the N-terminal catalytic cysteine, therefore mimicking the gamma-glutamyl-thioester intermediate formed during hydrolysis of glutamine. The fixation of the glutamine analog activates the enzyme through several major structural changes: 1) the closure of a loop to shield the glutaminase site accompanied by significant domain hinging, 2) the activation of catalytic residues involved in glutamine hydrolysis, i.e. the alpha-amino group of Cys-1 and Asn-98 that is positioned to form the oxyanion hole, and 3) a 75 degrees rotation of the Trp-74 indole group that opens the ammonia channel. PMID- 16339763 TI - A highly conserved amino-terminal region of sonic hedgehog is required for the formation of its freely diffusible multimeric form. AB - Although members of the Hedgehog (Hh) family were initially described as morphogens, many of these early conclusions were based on experiments that used non-physiologically relevant forms of Hh. Native Hh is modified by cholesterol (HhNp) and palmitate. These hydrophobic modifications are responsible for the ability of Hh to associate with cellular membranes, a property that initially appeared inconsistent with its ability to act far from its site of synthesis. Although it is now clear that Hh family members are capable of acting directly in long-range signaling, the form of Hh capable of this activity remains controversial. We have previously provided evidence for a freely diffusible multimeric form of Sonic Hedgehog (Shh) termed s-ShhNp, which is capable of accumulating in a gradient fashion through a morphogenic field. Here, we provide further evidence that s-ShhNp is the physiologically relevant form of Shh. We show that the biological activity of freely diffusible ShhNp resides in its multimeric form and that this multimeric form is exceedingly stable, even to high concentrations of salt and detergent. Furthermore, we now validate the Shh-Shh interactions previously observed in the crystal structure of human Shh, showing that a highly conserved amino-terminal domain of Shh is important for the formation of s-ShhNp. We also conclusively show that palmitoylation is required for s-ShhNp formation. Thus, our results identify both protein-protein and protein-lipid interactions that are required for s-ShhNp formation, and provide the first structural analyses supporting the existence of Shh multimers. PMID- 16339764 TI - p53 Mediates the accelerated onset of senescence of endothelial progenitor cells in diabetes. AB - Adverse metabolic factors, including oxidized small and dense low density lipoprotein (ox-dmLDL) can contribute to the reduced number and the impaired functions of circulating endothelial progenitors (EPC) in diabetic patients. To elucidate the molecular mechanisms involved, EPC from normal donors were cultured in the presence of ox-dmLDL. Under these experimental conditions EPC undergo to senescent-like growth arrest. This effect is associated with Akt activation, p21 expression, p53 accumulation, and retinoblastoma protein dephosphorylation and with a reduced protective effect against oxidative damage. Moreover, depletion of endogenous p53 expression by small interfering RNA demonstrates that the integrity of this pathway is essential for senescence to occur. Activation of the Akt/p53/p21 signaling pathway and accelerated onset of senescence are also detectable in EPC from diabetic patients. Finally, diabetic EPC depleted of endogenous p53 do not undergo to senescence-growth arrest and acquire the ability to form tube-like structures in vitro. These observations identify the activation of the p53 signaling pathway as a crucial event that can contribute to the impaired neovascularization in diabetes. PMID- 16339765 TI - Activation of Toll-like receptor 2 on microglia promotes cell uptake of Alzheimer disease-associated amyloid beta peptide. AB - The human G-protein-coupled formyl peptide receptor-like 1 (FPRL1) and its mouse homologue mFPR2 mediate the chemotactic activity of a variety of polypeptides associated with inflammation and bacterial infection, including the 42-amino acid form of amyloid beta peptide (Abeta42), a pathogenic factor in Alzheimer disease. Because mFPR2 was inducible in mouse microglial cells by proinflammatory stimulants, such as bacterial lipopolysaccharide, a ligand for the Toll-like receptor 4 (TLR4), we investigated the role of TLR2 in the regulation of mFPR2. We found that a TLR2 agonist, peptidoglycan (PGN) derived from Gram-positive bacterium Staphylococcus aureus, induced considerable mFpr2 mRNA expression in a mouse microglial cell line and primary microglial cells. This was associated with a markedly increased chemotaxis of the cells in response to mFPR2 agonist peptides. In addition, activation of TLR2 markedly enhanced mFPR2-mediated uptake of Abeta42 by microglia. Studies of the mechanistic basis showed that PGN activates MAPK and IkappaBalpha, and the effect of PGN on induction of mFPR2 was dependent on signaling pathways via ERK1/2 and p38 MAPKs. The use of TLR2 on microglial cells by PGN was supported by the fact that N9 cells transfected with short interfering RNA targeting mouse TLR2 failed to show increased expression of functional mFPR2 after stimulation with PGN. Our results demonstrated a potentially important role for TLR2 in microglial cells of promoting cell responses to chemoattractants produced in lesions of inflammatory and neurodegenerative diseases in the brain. PMID- 16339766 TI - The cysteine-rich secretory protein domain of Tpx-1 is related to ion channel toxins and regulates ryanodine receptor Ca2+ signaling. AB - The cysteine-rich secretory proteins (Crisp) are predominantly found in the mammalian male reproductive tract as well as in the venom of reptiles. Crisps are two domain proteins with a structurally similar yet evolutionary diverse N terminal domain and a characteristic cysteine-rich C-terminal domain, which we refer to as the Crisp domain. We presented the NMR solution structure of the Crisp domain of mouse Tpx-1, and we showed that it contains two subdomains, one of which has a similar fold to the ion channel regulators BgK and ShK. Furthermore, we have demonstrated for the first time that the ion channel regulatory activity of Crisp proteins is attributed to the Crisp domain. Specifically, the Tpx-1 Crisp domain inhibited cardiac ryanodine receptor (RyR) 2 with an IC(50) between 0.5 and 1.0 microM and activated the skeletal RyR1 with an AC(50) between 1 and 10 microM when added to the cytoplasmic domain of the receptor. This activity was nonvoltage-dependent and weakly voltage-dependent, respectively. Furthermore, the Tpx-1 Crisp domain activated both RyR forms at negative bilayer potentials and showed no effect at positive bilayer potentials when added to the luminal domain of the receptor. These data show that the Tpx-1 Crisp domain on its own can regulate ion channel activity and provide compelling evidence for a role for Tpx-1 in the regulation of Ca(2+) fluxes observed during sperm capacitation. PMID- 16339767 TI - Use of a dummy (pacifier) during sleep and risk of sudden infant death syndrome (SIDS): population based case-control study. AB - OBJECTIVES: To examine the association between use of a dummy (pacifier) during sleep and the risk of sudden infant death syndrome (SIDS) in relation to other risk factors. DESIGN: Population based case-control study. SETTING: Eleven counties in California. PARTICIPANTS: Mothers or carers of 185 infants whose deaths were attributed to SIDS and 312 randomly selected controls matched for race or ethnicity and age. MAIN OUTCOME MEASURE: Use of a dummy during sleep determined through interviews. RESULTS: The adjusted odds ratio for SIDS associated with using a dummy during the last sleep was 0.08 (95% confidence interval 0.03 to 0.21). Use was associated with a reduction in risk in every category of sociodemographic characteristics and risk factors examined. The reduced risk associated with use seemed to be greater with adverse sleep conditions (such as sleeping prone or on side and sleeping with a mother who smoked), although the observed interactions were not significant. In addition, use of a dummy may reduce the impact of other risk factors for SIDS, especially those related to adverse sleep environment. For example, infants who did not use a dummy and slept prone or on their sides (v on their back) had an increased risk of SIDS (2.61, 1.56 to 4.38). In infants who used dummies, there was no increased risk associated with sleeping position (0.66, 0.12 to 3.59). While cosleeping with a mother who smoked was also associated with increased risk of SIDS among infants who did not use a dummy (4.5, 1.3 to 15.1), there was no such association among those who did (1.1, 0.1 to 13.4). CONCLUSIONS: Use of a dummy seems to reduce the risk of SIDS and possibly reduces the influence of known risk factors in the sleep environment. PMID- 16339768 TI - Inhibitory mode of N-phenyl-4-pyrazolo[1,5-b] pyridazin-3-ylpyrimidin-2-amine series derivatives against GSK-3: molecular docking and 3D-QSAR analyses. AB - Glycogen synthase kinase 3 (GSK-3) inhibition is an important research topic because of its wide range of associated health implications. The interaction mode of a series of N-phenyl-4-pyrazolo[1,5-b]pyridazin-3-ylpyrimidin-2-amine compounds with human GSK-3 has been studied using molecular docking and 3D-QSAR approaches. In the 3D-QSAR studies, the molecular alignment and conformation determination are so important that they affect the success of a model. Flexible docking (AutoDock3.0.5) was used for the determination of 'active' conformation and molecular alignment. Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were used to develop 3D-QSAR models of 80 N-phenyl-4-pyrazolo[1,5-b]pyridazin-3-ylpyrimidin-2-amine compounds. The r(2) values were 0.870 and 0.861 for CoMFA and CoMSIA models, respectively. The predictive ability of these models was validated by 10 compounds of the test set. Mapping these models back to the topology of the active site of GSK-3 led to a better understanding of the vital N-phenyl-4 pyrazolo[1,5-b]pyridazin-3-ylpyrimidin-2-amines-GSK-3 interactions. The results demonstrate that combination of ligand-based and receptor-based modeling is a powerful approach to build 3D-QSAR models. The interaction mode from this study may be helpful for the design of a novel inhibitor and guide the selection of candidate sites for further experimental studies on site-directed mutagenesis. PMID- 16339769 TI - Floral nectar production and nectary anatomy and ultrastructure of Echinacea purpurea (Asteraceae). AB - BACKGROUND AND AIMS: In spite of the impressive species diversity in the Asteraceae and their widespread appeal to many generalist pollinators, floral nectary ultrastructure in the family has rarely been investigated. To redress this, a study using Echinacea purpurea, a plant of horticultural and nutraceutical value, was undertaken. Nectar secretion of disc florets was compared with floral nectary ultrastructure taking into account nectar's potential impact upon the reproductive success of this outcrossing species. METHODS: Micropipette collections of nectar in conjunction with refractometry were used to determine the volume and nectar-sugar quantities of disc florets throughout their phenology, from commencement of its production to cessation of secretion. Light, scanning-electron and transmission-electron microscopy were utilized to examine morphology, anatomy and ultrastructure of nectaries of the disc florets. KEY RESULTS: Florets were protandrous with nectar being secreted from anthesis until the third day of the pistillate phase. Nectar production per floret peaked on the first day of stigma receptivity, making the two innermost whorls of open florets most attractive to foraging visitors. Modified stomata were situated along the apical rim of the collar-like nectary, which surrounds the style base and sits on top of the inferior ovary. The floral nectary was supplied by phloem only, and both sieve elements and companion cells were found adjacent to the epidermis; the latter participated in the origin of some of the precursor cells that yielded these specialized cells of phloem. Companion cells possessed wall ingrowths (transfer cells). Lobed nuclei were a key feature of secretory parenchyma cells. CONCLUSIONS: The abundance of mitochondria suggests an eccrine mechanism of secretion, although dictyosomal vesicles may contribute to a granulocrine process. Phloem sap evidently is the main contributor of nectar carbohydrates. From the sieve elements and companion cells, an apoplastic route via intercellular spaces and cell walls, leading to the pores of modified stomata, is available. A symplastic pathway, via plasmodesmata connecting sieve elements to companion, parenchyma and epidermal cells, is also feasible. Uncollected nectar was reabsorbed, and the direct innervation of the nectary by sieve tubes potentially serves a second important route for nectar-sugar reclamation. Microchannels in the outer cuticle may facilitate both secretion and reabsorption. PMID- 16339771 TI - Why do general practitioners prescribe antibiotics for acute infective conjunctivitis in children? Qualitative interviews with GPs and a questionnaire survey of parents and teachers. AB - BACKGROUND: Acute infective conjunctivitis in children is a common presentation in primary care. Treatment is usually with antibiotics and prescribing may be affected by non-clinical factors. AIMS: To investigate the non-clinical determinants of the management of acute infective conjunctivitis in children. DESIGN: Qualitative interviews with GPs and a questionnaire survey of parents of children with acute infective conjunctivitis and teachers. SETTING: GPs in Sheffield and Berkshire and parents of children with acute infective conjunctivitis and schools in Oxfordshire. METHODS: Semi-structured telephone interviews of 39 GPs. Questionnaire survey of 326 parents of children enrolled into a trial of acute infective conjunctivitis treatment. Questionnaire survey of 223 nurseries and primary schools in Oxfordshire. RESULTS: All three groups agreed that acute infective conjunctivitis was a mild condition. Parents were certain about the benefits of antibiotic treatment and sought early consultations with their GP in a desire to get their child back to school. GPs sometimes collude with a parent's request to prescribe to enable school attendance. Despite this 54.2% (95%CI 48.5-59.8%) children missed a mean of 1.85 days from school and 28.6% of parents (95%CI 23.5-33.7%) missed a mean of 1.5 days off work. CONCLUSION: Social factors, including the need for children to attend day care or school and parents to go to work, contribute to the decision to prescribe antibiotics for children with acute infective conjunctivitis. Understanding these issues and changing school policies in line with national guidance may reduce pressure on GPs to prescribe for this condition. PMID- 16339770 TI - Nitrogen regulation of root branching. AB - BACKGROUND: Many plant species can modify their root architecture to enable them to forage for heterogeneously distributed nutrients in the soil. The foraging response normally involves increased proliferation of lateral roots within nutrient-rich soil patches, but much remains to be understood about the signalling mechanisms that enable roots to sense variations in the external concentrations of different mineral nutrients and to modify their patterns of growth and development accordingly. SCOPE: In this review we consider different aspects of the way in which the nitrogen supply can modify root branching, focusing on Arabidopsis thaliana. Our current understanding of the mechanism of nitrate stimulation of lateral root growth and the role of the ANR1 gene are summarized. In addition, evidence supporting the possible role of auxin in regulating the systemic inhibition of early lateral root development by high rates of nitrate supply is presented. Finally, we examine recent evidence that an amino acid, L-glutamate, can act as an external signal to elicit complex changes in root growth and development. CONCLUSIONS: It is clear that plants have evolved sophisticated pathways for sensing and responding to changes in different components of the external nitrogen supply as well as their own internal nitrogen status. We speculate on the possibility that the effects elicited by external L glutamate represent a novel form of foraging response that could potentially enhance a plant's ability to compete with its neighbours and micro-organisms for localized sources of organic nitrogen. PMID- 16339772 TI - Two approaches to school health promotion--a focus on health-related behaviours and general competencies. An ecological study of 25 Swedish municipalities. AB - The school is an important potential health-promoting setting for children and adolescents. Two main perspectives on school health promotion have been identified, one addressing health-related behaviours, the other stressing the development of general competencies. From a policy perspective, it is important to establish whether these two approaches are complementary or competing. This question was analysed by examining school administrations in 25 municipalities in Stockholm County, Sweden. An attempt was made to relate level of activity in line with each approach to general adolescent health outcomes. Data related to the two approaches were collected through the administration of a questionnaire. Outcome variables, measured as fraction of students qualified for upper secondary school, fraction of students with high alcohol intake, and fraction of 15-18 year olds suspected of crime, were measured on the basis of registry data. To control for structural patterns, socio-economic, demographic and system characteristics were gathered from public statistics. Spearman rank correlation coefficients were computed. Clusters of determinants assignable to each of the two approaches- promotion of health-related behaviours and supporting the development of student competencies--were identified. The two approaches were found to be negatively correlated. Thus, the perspective focusing on promotion of student health-related behaviours and the one emphasizing general development of student competencies seem to be competing rather than complementary. If this result is confirmed by other studies, it will have important policy implications. PMID- 16339773 TI - Participation rates in worksite-based intervention studies: health promotion context as a crucial quality criterion. AB - Recently, a set of specific quality evaluation criteria for health promotion research has been proposed in this journal. One of the quality criteria identified is the 'health promotion context'. With this paper we would like to contribute to the dialogue by specifying the importance of this criterion on the basis of our on experience with worksite-based obesity prevention interventions. We advocate the reporting of participation rates among approached worksites in publications on worksite intervention effects. Such information will help to draw conclusions on the practical relevance of the shown effectiveness of the intervention. Health promotion practice is advised to adopt and disseminate evidence-based interventions, accompanied by a diffusion study with a minimal research burden for participants. PMID- 16339774 TI - Healthy settings: challenges to generating evidence of effectiveness. AB - This paper starts by briefly reviewing the history, theory and practice of the settings approach to promoting public health--highlighting its ecological perspective, its understanding of settings as dynamic open systems and its primary focus on whole system organization development and change. It goes on to outline perceived benefits and consider why, almost 20 years after the Ottawa Charter advocated the approach, there remains a relatively poorly developed evidence base of effectiveness. Identifying three key challenges--relating to the construction of the evidence base for health promotion, the diversity of conceptual understandings and real-life practice and the complexity of evaluating ecological whole system approaches--it suggests that these have resulted in an ongoing tendency to evaluate only discrete projects in settings, thus failing to capture the 'added value' of whole system working. It concludes by exploring the potential value of theory-based evaluation and identifying key issues that will need to be addressed in moving forward--funding evaluation within and across settings; ensuring links between evidence, policy and practice; and clarifying and articulating the theories that underpin the settings approach generically and inform the approach as applied within particular settings. PMID- 16339775 TI - Theory-based survey analysis of well-being in secondary schools in Finland. AB - Frameworks concerning school health and well-being, like the Health Promoting School and the Coordinated School Health Programme, have been available for some time. However, the indicators of well-being studies and the comprehensive frameworks have not met. The main aim of the present study was to test the use of the theoretically based School Well-being Profile to clarify the aspects of well being in schools and to test if the Profile could be used to identify differences in well-being between classes in school. The data were collected in connection with the Koulumiete Project in Finland; 1346 pupils and 69 classes from grades seven to nine were included in the present data. In the school conditions category, the physical conditions that most needed improvement were ventilation, toilet facilities and temperature. Time pressure was also reported widely. Concerning social relationships, teachers' interest in how pupils are doing and teachers' fair treatment were the aspects calling for attention. In the means for self-fulfillment category pupils' participation in school development was a key area for reform. The most prevalent psychosomatic symptoms in the health status category were headache and feeling tired or weak. The School Well-being Profile seemed to work well when evaluating well-being differences between classes. By utilizing the results locally, the school personnel can act to develop their own school. The Profile could be used as a tool when planning and evaluating well being and mental health promotion interventions in schools. PMID- 16339776 TI - Variant progesterone receptor mRNAs are co-expressed with the wild-type progesterone receptor mRNA in human endometrium during all phases of the menstrual cycle. AB - Progesterone receptor (PR) variant mRNAs in human endometrium could encode proteins with the potential to alter progesterone action in states of normal and abnormal endometrial development. We have assessed the expression levels of mRNA for the wild-type PR and splice variants of PR mRNA lacking exon 4 (del-4 PR), exon 6 (del-6 PR), exons 4 and 6 (del-4&6 PR), and part of exon 4 (del-p4 PR) or part of exon 6 (del-p6 PR) in the human endometrium throughout menstrual cycle development. Eighty-eight endometrial specimens (47 proliferative, 41 secretory) were collected from patients undergoing hysterectomy for benign gynaecologic causes. Measurements by RT-PCR indicated that mRNAs for wild-type PR, and splice variants del-4 PR, del-6 PR, del-4&6 PR, del-p6 PR, and a novel del-p4 PR were detected in all endometrial specimens throughout the menstrual cycle. Higher levels of wild-type PR and all PR variant mRNAs were found in the early and mid proliferative endometrial phases than in secretory endometrium. The relative expression of mRNA for all PR variants compared to wild-type PR mRNA, however, did not change through all stages of endometrial development. We, therefore, found no evidence of differential co-expression of the PR variants compared with wild-type PR during normal menstrual development. Future studies will determine if the expression profile of PR variant mRNAs will be different in the endometrium of patients with infertility, recurrent pregnancy loss, or endometrial adenocarcinoma. PMID- 16339777 TI - Transforming growth factor beta1 regulates angiotensin II type I receptor gene expression in the extravillous trophoblast cell line SGHPL-4. AB - The angiotensin II type 1 (AT1) receptor, transforming growth factor beta1 (TGFbeta1) and Oncostatin M (OSM) control key pathways that may be important during placentation. Although interactions between them exist in other tissues, trophoblast cells have not been investigated. Extravillous trophoblast cells, SGHPL-4, were stimulated with 10 ng/ml TGFbeta1 +/- 100 ng/ml OSM for 24 h. Real time PCR showed that AT1 expression increased 2.76-fold [95% confidence interval (CI) = 1-6.74, P = 0.05] in response to TGFbeta1 and 4.21-fold (95% CI = 1.33 11.76, P = 0.03) with TGFbeta1 + OSM. Luciferase reporter gene constructs containing three haplotypes of the 59 flanking region of the AT1 receptor gene were transfected into SGHPL-4 and HepG2 cells and stimulated with 0.1, 1 and 10 ng/ml TGFbeta1 and 50 ng/ml OSM. Responses were dose and cell dependent. Luciferase activity increased in HepG2 cells in response to TGFbeta1 alone or together with OSM (P < 0.001); transcriptional activation differed between AT1 receptor gene haplotypes. In SGHPL-4 cells, luciferase activity was reduced on exposure to low concentrations of TGFbeta1 or high concentrations of TGFbeta1 combined with OSM (P = 0.003); the response was unaffected by haplotype. Interaction between AT1 and TGFbeta1 is a novel observation in trophoblast and suggests new avenues for the study of placentation. PMID- 16339778 TI - Role of protein kinase G in barrier-protective effects of cGMP in human pulmonary artery endothelial cells. AB - Increases in endothelial cGMP prevent oxidant-mediated endothelial barrier dysfunction, but the downstream mechanisms remain unclear. To determine the role of cGMP-dependent protein kinase (PKG)(I), human pulmonary artery endothelial cells (HPAEC) lacking PKG(I) expression were infected with a recombinant adenovirus encoding PKG(Ibeta) (Ad.PKG) and compared with uninfected and control infected (Ad.betagal) HPAEC. Transendothelial electrical resistance (TER), an index of permeability, was measured after H(2)O(2) (250 microM) exposure with or without pretreatment with 8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphate (CPT-cGMP). HPAEC infected with Ad.PKG, but not Ad.betagal, expressed PKG(I) protein and demonstrated Ser(239) and Ser(157) phosphorylation of vasodilator-stimulated phosphoprotein after treatment with CPT-cGMP. Adenoviral infection decreased basal permeability equally in Ad.PKG- and Ad.betagal-infected HPAEC compared with uninfected cells. Treatment with CPT-cGMP (100 microM) caused a PKG(I)-independent decrease in permeability (8.2 +/- 0.6%). In all three groups, H(2)O(2) (250 microM) caused a similar approximately 35% increase in permeability associated with increased actin stress fiber formation, intercellular gaps, loss of membrane VE-cadherin, and increased intracellular Ca(2+) concentration ([Ca(2+)](i)). In uninfected and Ad.betagal-infected HPAEC, pretreatment with CPT-cGMP (100 microM) partially blocked the increased permeability induced by H(2)O(2). In Ad.PKG-infected HPAEC, CPT-cGMP (50 microM) prevented the H(2)O(2)-induced TER decrease, cytoskeletal rearrangement, and loss of junctional VE-cadherin. CPT-cGMP attenuated the peak [Ca(2+)](i) caused by H(2)O(2) similarly (23%) in Ad.betagal- and Ad.PKG-infected HPAEC, indicating a PKG(I)-independent effect. These data suggest that cGMP decreased HPAEC basal permeability by a PKG(I)-independent process, whereas the ability of cGMP to prevent H(2)O(2)-induced barrier dysfunction was predominantly mediated by PKG(I) through a Ca(2+)-independent mechanism. PMID- 16339779 TI - CXCR3 surface expression in human airway epithelial cells: cell cycle dependence and effect on cell proliferation. AB - We recently demonstrated that human bronchial epithelial cells (HBEC) constitutively express the CXC chemokine receptor CXCR3, which when activated, induces directed cell migration. The present study in HBEC examined the relative expression of the CXCR3 splice variants CXCR3-A and -B, cell cycle dependence of CXCR3 expression, and the effects of the CXCR3 ligand, the interferon-gamma inducible CXC chemokine I-TAC/CXCL11, on DNA synthesis and cell proliferation. Both CXCR3-A and -B mRNA, assessed by real-time RT-PCR, were expressed in normal HBEC (NHBEC) and the HBEC line 16-HBE. However, CXCR3-B mRNA was 39- and 6-fold greater than CXCR3-A mRNA in NHBEC and 16-HBE, respectively. Although most HBEC (>80%) assessed by flow cytometry and immunofluorescence microscopy contained intracellular CXCR3, only a minority (<40%) expressed it on the cell surface. In this latter subset of cells, most (>75%) were in the S + G(2)/M phases of the cell cycle. Stimulation of CXCR3 with I-TAC enhanced thymidine incorporation and cell proliferation and increased p38 and ERK1/2 phosphorylation. These data indicate that 1) human airway epithelial cells primarily express CXCR3-B mRNA, 2) surface expression of CXCR3 is largely confined to the S + G(2)/M phases of the cell cycle, and 3) activation of CXCR3 induces DNA synthesis, cell proliferation, and activation of MAPK pathways. We speculate that activation of CXCR3 exerts a mitogenic effect in HBEC, which may be important during airway mucosal injury in obstructive airway diseases such as asthma and chronic obstructive pulmonary disease. PMID- 16339780 TI - Effect of adenosine A2A receptor activation in murine models of respiratory disorders. AB - Activation of the adenosine A(2A) receptor has been postulated as a possible treatment for lung inflammatory diseases such as asthma and chronic obstructive pulmonary disease (COPD). In this report, we have studied the anti-inflammatory properties of the reference A(2A) agonist CGS-21680, given intranasally at doses of 10 and 100 microg/kg, in a variety of murine models of asthma and COPD. After an acute ovalbumin challenge of sensitized mice, prophylactic administration of CGS-21680 inhibited the bronchoalveolar lavage fluid inflammatory cell influx but not the airway hyperreactivity to aerosolized methacholine. After repeated ovalbumin challenges, CGS-21680 given therapeutically inhibited the bronchoalveolar lavage fluid inflammatory cell influx but had no effect on the allergen-induced bronchoconstriction, the airway hyperreactivity, or the bronchoalveolar lavage fluid mucin levels. As a comparator, budesonide given intranasally at doses of 0.1-1 mg/kg fully inhibited all the parameters measured in the latter model. In a lipopolysaccharide-driven model, CGS-21680 had no effect on the bronchoalveolar lavage fluid inflammatory cell influx or TNF-alpha, keratinocyte chemoattractant, and macrophage inflammatory protein-2 levels, but potently inhibited neutrophil activation, as measured by bronchoalveolar lavage fluid elastase levels. With the use of a cigarette smoke model of lung inflammation, CGS-21680 did not significantly inhibit bronchoalveolar lavage fluid neutrophil infiltration but reversed the cigarette smoke-induced decrease in macrophage number. Together, these results suggest that activation of the A(2A) receptor would have a beneficial effect by inhibiting inflammatory cell influx and downregulating inflammatory cell activation in asthma and COPD, respectively. PMID- 16339781 TI - Neutrophil transendothelial migration in vitro to Streptococcus pneumoniae is pneumolysin dependent. AB - The recruitment of polymorphonuclear leukocytes (PMN) from the vascular space to the alveolar air space is an early event in host defense against pneumococcal pneumonia. Pneumolysin is a virulence factor for Streptococcus pneumoniae, but a specific role for pneumolysin in neutrophil-endothelial cell interactions has not been investigated. Using a Transwell system, we studied in vitro migration of PMNs across a monolayer of human pulmonary microvascular endothelial cells in response to wild-type S. pneumoniae (D39) and a pneumolysin-deficient mutant (plnA(-)) incubated on the abluminal surface of the monolayer. S. pneumoniae induction of PMN migration was dose dependent and elicited by > or =10(5) D39. Mutants lacking pneumolysin had dramatically reduced potency for eliciting PMN migration compared with the parent strain (5 x 10(6) plnA(-) elicits 18.6% PMN migration vs. 55.5% for 5 x 10(6) D39). The disparity between D39 and plnA(-) persisted in ethanol-fixed bacteria, consistent with the properties of pneumolysin. Neither conditioned medium from D39 nor purified pneumolysin elicited PMN migration to the same extent as the intact D39, suggesting that the role of pneumolysin in eliciting PMN migration requires a more complex interaction between the organism, the endothelium, and the PMN. Both D39 and plnA(-) adhered to, and translocated across, the endothelium in the abluminal to luminal direction and elicited similar levels of IL-8 production. Neither strain elicited upregulation of the endothelial adhesion molecules ICAM-1, VCAM-1, or E selectin, and they did not cause translocation of NF-kappaB to the nucleus. These findings demonstrate a novel role for pneumolysin in pneumococcus-induced PMN recruitment across the pulmonary endothelium. PMID- 16339782 TI - BMP signaling controls PASMC KV channel expression in vitro and in vivo. AB - Bone morphogenetic proteins (BMPs) have been implicated in the pathogenesis of familial pulmonary arterial hypertension. The type 2 receptor (BMPR2) is required for recognition of all BMPs. Transgenic mice with a smooth muscle cell-targeted mutation in this receptor (SM22-tet-BMPR2(delx4+)) developed increased pulmonary artery pressure, associated with a modest increase in arterial muscularization, after 8 wk of transgene activation (West J, Fagan K, Steudel W, Fouty B, Lane K, Harral J, Hoedt-Miller M, Tada Y, Ozimek J, Tuder R, and Rodman DM. Circ Res 94: 1109-1114, 2004). In the present study, we show that these transgenic mice developed increased right ventricular pressures after only 1 wk of transgene activation, without significant remodeling of the vasculature. We then tested the hypothesis that the increased pulmonary artery pressure due to loss of BMPR2 signaling was mediated by reduced K(V) channel expression. There was decreased expression of K(V)1.1, K(V)1.5, and K(V)4.3 mRNA isolated from whole lung. Western blot confirmed decreased K(V)1.5 protein in these lungs. Human pulmonary artery smooth muscle cells (PASMC) treated with recombinant BMP2 had increased K(V)1.5 protein and macroscopic K(V) current density, which was blocked by anti K(V)1.5 antibody. In vivo, nifedipine, a selective L-type Ca(2+) channel blocker, reduced RV systolic pressure in these dominant-negative BMPR2 mice to levels seen in control animals. This suggests that activation of L-type Ca(2+) channels caused by reduced K(V)1.5 mediates increased pulmonary artery pressure in these animals. These studies suggest that BMP regulates K(V) channel expression and that loss of this signaling pathway in PASMC through a mutation in BMPR2 is sufficient to cause pulmonary artery vasoconstriction. PMID- 16339783 TI - Imaging photosynthesis in wounded leaves of Arabidopsis thaliana. AB - Chlorophyll fluorescence imaging provides a non-invasive and non-destructive means with which to measure photosynthesis. This technique has been used, in combination with 14CO2 feeding, to study the spatial and temporal changes in source-sink relationships which occur in mechanically wounded leaves of Arabidopsis thaliana. Twenty-four hours after wounding, cells proximal to the wound margin showed a rapid induction of PhiII upon illumination (a measure of the efficiency of photosystem II photochemistry) whilst cells more distal to the wound margin exhibited a much slower induction of PhiII and a large, transient increase in NPQ (a measure of the rate constant for non-photochemical energy dissipation within the light-harvesting antenna). These results are indicative of an increase in sink strength in the vicinity of the wound and this was confirmed by the retention of 14C photosynthate in this region. It has been hypothesized that wound-induced cell wall (apoplastic) invertase (cwINV) activity plays a central role in generating localized increases in sink strength in stressed plant tissue and that hexose sugars generated by the sucrolytic activity of cwINV may act as a signal regulating gene expression. Enzyme activity measurements, quantitative RT-PCR, and T-DNA insertional mutagenesis have been used to determine that expression of AtcwINV1 is responsible for all induced cwINV activity in mechanically wounded leaves. Whilst inactivation of this gene abolished wound-induced cwINV activity, it did not affect localized alterations in source-sink relationships of wounded leaves or wound-regulated gene expression. The signals that may regulate source-sink relationships and signalling in wounded leaves are discussed. PMID- 16339784 TI - Role of abscisic acid (ABA) and Arabidopsis thaliana ABA-insensitive loci in low water potential-induced ABA and proline accumulation. AB - The mechanisms by which plants respond to reduced water availability (low water potential) include both ABA-dependent and ABA-independent processes. Pro accumulation and osmotic adjustment are two important traits for which the mechanisms of regulation by low water potential, and the involvement of ABA, is not well understood. The ABA-deficient mutant, aba2-1, was used to investigate the regulatory role of ABA in low water potential-induced Pro accumulation and osmotic adjustment in seedlings of Arabidopsis thaliana. Low water potential induced Pro accumulation required wild-type levels of ABA, as well as a change in ABA sensitivity or ABA-independent events. Osmotic adjustment, in contrast, occurred independently of ABA accumulation in aba2-1. Quantification of low water potential-induced ABA and Pro accumulation in five ABA-insensitive mutants, abi1 1, abi2-1, abi3, abi4, and abi5, revealed that abi4 had increased Pro accumulation at low water potential, but a reduced response to exogenous ABA. Both of these responses were modified by sucrose treatment, indicating that ABI4 has a role in connecting ABA and sugar in regulating Pro accumulation. Of the other abi mutants, only abi1 had reduced Pro accumulation in response to low water potential and ABA application. It was also observed that abi1-1 and abi2-1 had increased ABA accumulation. The involvement of these loci in feedback regulation of ABA accumulation may occur through an effect on ABA catabolism or conjugation. These data provide new information on the function of ABA in seedlings exposed to low water potential and define new roles for three of the well-studied abi loci. PMID- 16339785 TI - Polymorphism and modulation of cell wall esterase enzyme activities in the chicory root during the growing season. AB - Pectins are major components of the primary plant cell wall. They can be both methylesterified and acetylesterified and de-esterification occurs by specific esterases. Proteins extracted by NaCl treatment from root cell walls of two chicory varieties (Cichorium intybus L. cv. Nausica and Arancha) sampled in an experimental field every 2 weeks between July 2002 and January 2003 were analysed by isoelectrofocalization, semi-denaturing SDS-PAGE, and quantitative assays for their esterase activity. Zymograms showed that chicory root pectin methylesterases belong to a multigene family. The isoelectric points of the pectin methylesterase isoforms ranged from pI 3.8 to pI 9.0. Concerning acetylesterases, only acidic isoforms between pI 4.1 and pI 5.2 were observed, but a large polymorphism of this class of enzymes could be identified in one variety. The results indicate that the root pectin methylesterase activity of the Nausica variety was correlated with ambient temperature, while no significant effect of temperature could be detected on any acetylesterase isoform. PMID- 16339786 TI - Prescribing practices of doctors attending to under fives in a children's outpatient clinic in Owerri, Nigeria. AB - Prescribing practices of doctors in different clinical settings have been documented but there is a dearth of information on prescribing practices with regards to children especially the under fives. This study set to describe the prescribing practices of doctors attending to under fives in a children's outpatient clinic. The information obtained is expected to aid in designing appropriate interventions. Between January and April 2004 the age, sex and drugs prescribed for under fives seen at the children's outpatient clinic of Federal medical centre, Owerri in South eastern Nigeria were extracted at the end of each day's consultations from their cards and entered into a spread sheet. Patients who had no prescriptions were excluded. A total of 2471 medications were prescribed for 790 patients who met the criteria for inclusion in the study. Antimalarials, Analgesics, Antibiotics, Vitamin C, Antihistamines and Multivitamin preparations were the commonest drugs prescribed. The prescription rate per patient was 3.13. While three different antimalarials were prescribed a total of twelve different antibiotics were. Prescription rate for injections was 1.9 per cent. Only 13.3 per cent of the patients had all their drugs prescribed in generic names. The others had at least two drugs prescribed in brand names. The difference in cost between same drugs prescribed in brand names as against in generic names were between 41.7 per cent and 60 per cent. All the antimalarials and analgesics prescribed were in the Nigerian National essential drug list while only 16.7 per cent of antibiotics prescribed were not. This study has documented significant flaws in the prescribing practices of these doctors, particularly the low rate of prescription in generic names, high rate of antibiotics prescription, inappropriate prescription of multivitamin preparations and Vitamin C and a relatively high rate of poly pharmacy. Suggested interventions include developing and circulating easy to use treatment guidelines for diseases commonly seen in our centre and a regular audit of the application of these guidelines. Continuing medical education of doctors on rational drug use and evidence based medicine should also be instituted. PMID- 16339787 TI - Nitric oxide and reactive oxygen species production causes progressive damage in rats after cessation of silica inhalation. AB - Our laboratory has previously reported results from a rat silica inhalation study which determined that, even after silica exposure ended, pulmonary inflammation and damage progressed with subsequent fibrosis development. In the present study, the relationship between silica exposure, nitric oxide (NO) and reactive oxygen species (ROS) production, and the resultant pulmonary damage is investigated in this model. Rats were exposed to silica (15 mg/m3, 6 h/day) for either 20, 40, or 60 days. A portion of the rats from each exposure were sacrificed at 0 days postexposure, while another portion was maintained without further exposure for 36 days to examine recovery or progression. The major findings of this study are: (1) silica-exposed rat lungs were in a state of oxidative stress, the severity of which increased during the postexposure period, (2) silica-exposed rats had significant increase in lung NO production which increased in magnitude during the postexposure period, and (3) the presence of silica particle(s) in an alveolar macrophage (AM) was highly associated with inducible nitric oxide synthase (iNOS) protein. These data indicate that, even after silica exposure has ended, and despite declining silica lung burden, silica-induced pulmonary NO and ROS production increases, thus producing a more severe oxidative stress. A quantitative association between silica and expression of iNOS protein in AMs was also determined, which adds to our previous observation that iNOS and NO-mediated damage are associated anatomically with silica-induced pathological lesions. Future studies will be needed to determine whether the progressive oxidative stress, and iNOS activation and NO production, is a direct result of silica lung burden or a consequence of silica-induced biochemical mediators. PMID- 16339788 TI - Assessing the potential to induce respiratory hypersensitivity. AB - Acute and repeat dose inhalation studies have been an important part of the safety assessment of drugs, chemicals, and other products throughout the world for many years. It is known that damage to the respiratory tract can be triggered either by nonspecific irritation or by specific immune-mediated pathogenesis, and it is acknowledged that traditional inhalation studies are not designed to address fully the impact of the latter. It is also recognized that different types of immune-mediated responses can be triggered by different classes of compounds and that some immune reactions in the lung are life threatening. As such, it is important to understand as fully as possible the basis for the immune mediated damage to the lung in order to characterize adequately the risks of individual chemicals or proteins. It is against this background that a review of the methods used to assess the potential for immune-mediated respiratory hypersensitivity was conducted. The primary objectives of this review are to discuss appropriate methods for identifying and characterizing respiratory hypersensitivity hazards and risks; and to identify key data gaps and related research needs with respect to respiratory hypersensitivity testing. The following working definition of respiratory hypersensitivity was formulated: a hypersensitivity response in the respiratory tract precipitated by a specific immune response, mediated by multiple mechanisms, including IgE antibody. Because of the importance played by various classes of compounds, the subsequent sections of this review will consider protein-specific, chemical-specific, and drug specific aspects of respiratory hypersensitivity. PMID- 16339789 TI - Effect of PCB 126 on hepatic metabolism of thyroxine and perturbations in the hypothalamic-pituitary-thyroid axis in the rat. AB - The objective of this research was to examine the time- and dose- dependent disturbances in the hypothalamic-pituitary-thyroid (HPT) axis of adult male rats administered a potent coplanar (non-ortho) PCB, 3,3',4,4',5-pentachlorobiphenyl (PCB 126). Adult male Sprague-Dawley rats were administered a single oral bolus dose of 0, 7.5, 75, or 275 microg PCB 126/kg bw dissolved in corn oil. The rats were sacrificed periodically over 22 days. The 7.5-microg/kg dose induced hepatic ethoxyresorufin-O-deethylation EROD activity, but no changes were observed in hepatic uridine diphosphate glucuronyl transferases (UDPGTs) activity or serum TSH, T4, or fT4 concentrations. The two highest doses caused a modest decline in weight gain, induced hepatic EROD and UDPGT activities, increased serum TSH concentrations, and decreased serum T4 and fT4 concentrations. The amount of thyroxine glucuronide formed daily (pM/mg protein) increased linearly with the area-under-the-concentration-curve (AUCC) for PCB 126 in liver (microg/kg/day) and then slowed at the 275-microg/kg PCB 126 dose. Perturbations in the HPT axis were nonlinear with respect to PCB 126 dosing. As expected, an inverse relationship between the AUCC for serum T4 (microg/dl/day) and the AUCC for serum TSH (ng/dl/day) was observed; however, the relationship was highly nonlinear. These data support a mode of action for PCB 126 involving induction of hepatic UDPGTs by the aryl hydrocarbon receptor AhR. However, the dose-response characteristics of the HPT axis are nonlinear and complex, requiring sophisticated tools, such as PBPK models, to characterize dose response. PMID- 16339790 TI - Changes in the effect of propofol in response to altered plasma protein binding during normothermic cardiopulmonary bypass. AB - BACKGROUND: During normothermic cardiopulmonary bypass (CPB), the effect on propofol pharmacokinetics of changes in its binding to plasma proteins is consistent with the predictions of the well-stirred model of hepatic elimination for nonrestrictively cleared drug. However, whether changes in binding lead to clinically significant changes in the drug effect remains unclear. The purpose of this study was to assess changes in the drug effect of propofol in response to altered plasma binding using quantitative EEG measurements. METHODS: Thirty patients undergoing cardiac surgery were assigned randomly to receive propofol infusions at 4 (Group P-4) or 6 (Group P-6) mg kg(-1) h(-1) during surgery. The concentration of propofol in blood samples, collected from the radial artery at predetermined intervals, was determined by HPLC. The unbound fraction of drug in plasma was estimated using equilibrium dialysis. Bispectral index (BIS) and burst suppression ratio (BSR) were measured at the time blood samples were collected. RESULTS: The total concentration of propofol in blood was unchanged during CPB relative to the pre-CPB value in both groups. However, the fraction of unbound propofol in blood increased by 2-fold during CPB. While BIS values were unchanged during CPB in Group P-4, there was a slight, but significant, decrease in Group P 6. In both groups, BSR significantly increased during CPB. BIS values showed a weak correlation with the concentration of unbound propofol (r(2)=0.19, P<0.001). BSR showed a moderate correlation with the concentration of unbound propofol (r(2)=0.56, P<0.001). CONCLUSIONS: The anaesthetic effect of propofol significantly increased during CPB without any alteration in the total drug concentration. The enhanced efficacy may be caused by a reduction in plasma binding of the drug. PMID- 16339791 TI - Intra-articular injection of warmed lidocaine improves intraoperative anaesthetic and postoperative analgesic conditions. AB - BACKGROUND: Although local anaesthesia for knee arthroscopy is a well-documented procedure, arthroscopy under local anaesthesia is often interrupted because of intolerable discomfort and pain. Warming local anaesthetic solutions may increase its anaesthetic effect. We tested whether intra-articular injection of warmed lidocaine solution could improve intraoperative anaesthetic and postoperative analgesic conditions. METHODS: Patients in the warmed group received 20 ml warmed (40 degrees C) lidocaine 1% intra-articularly 20 min before surgery. The patients in the control group received 20 ml room-temperature (25 degrees C) lidocaine 1% intra-articularly 20 min before surgery. During surgery, the patients reported pain on a visual analogue scale (VAS). RESULTS: The median VAS pain score was 1.5 (range, 0.0-3.0) in the warmed lidocaine group and 5.0 (4.0-8.0) in the control group (P<0.001). The median intra- and postoperative analgesic requirements in the control group were significantly greater than that in the warmed group. CONCLUSION: Warmed lidocaine injected intra-articularly provides improved intraoperative anaesthetic and postoperative analgesic conditions for patients undergoing knee arthroscopy. PMID- 16339792 TI - Effects of osteoprotegerin administration on osteoclast differentiation and trabecular bone structure in osteoprotegerin-deficient mice. AB - Osteoprotegerin (OPG)-deficient mice exhibit severe bone loss including the destruction of growth plate cartilage. Using OPG-deficient mice, we attempted to clarify the differentiation and ultrastructure of osteoclasts located on the destroyed growth plate cartilage and trabecular bone matrix in long bones. In (-/ ) homozygous OPG knockout mice, adjacent to the growth plate cartilage, the formation of bone trabeculae without a calcified cartilaginous core resulted in an irregular chondrocyte distribution in the growth plate cartilage. At the metaphyseal ossification center, TRAP-positive osteoclasts showed unusual localization on both type-II collagen-positive cartilage and type-I collagen positive bone matrix. Osteoclasts located on cartilage matrix lacked a typical ruffled border structure, but formed resorption lacunae. During growth plate cartilage destruction, osteoclasts formed ruffled border structures on bone matrix deposited on the remaining cartilage surfaces. These findings suggest that, in OPG (-/-) mice, osteoclast structure differs, depending on the matrix of either cartilage or bone. Then, we examined the effects of OPG administration on the internal trabecular bone structure and osteoclast differentiation in OPG (-/ ) mice. OPG administration to OPG (-/-) mice significantly inhibited trabecular bone loss and maintained the internal trabecular bone structure, but did not reduce the osteoclast number on bone trabeculae. For most osteoclasts, OPG administration caused disappearance or reduction of the ruffled border, but induced neither necrotic nor apoptotic damages. These results suggest that OPG administration is an effective means of maintaining the internal structure and volume of trabecular bone in metabolic bone diseases by inhibition of osteoclastic bone resorption. PMID- 16339793 TI - Grey matter heterotopia: what EEG-fMRI can tell us about epileptogenicity of neuronal migration disorders. AB - Grey matter heterotopia are commonly associated with refractory epilepsy. Depth electrodes recordings have shown that epileptiform activity can be generated within these lesions, and also at a distance in the neocortex. Heterotopia seem to be part of a more complex circuitry involving also the surrounding and distant cerebral cortex. Blood oxygenation level-dependent (BOLD) changes to interictal spikes using continuous EEG and functional MRI (EEG-fMRI) can help to understand non-invasively the mechanisms of epileptogenicity in these patients. We studied 14 patients with epilepsy and heterotopia using simultaneous recording of EEG fMRI. EEG was continuously acquired from inside the scanner during 2 h sessions. Epileptic spikes were visually identified in the filtered EEG and each type of spike determined one EEG-fMRI study. We looked at positive (activation) and negative (deactivation) changes in the BOLD signal. Eleven patients had nodular heterotopia and three band heterotopia. Four patients had more than one type of spikes, with a total of 26 EEG-fMRI studies. We excluded three with less than three spikes, and therefore a total of 23 studies (12 with nodular and 11 with band heterotopia) were analysed. Nodular heterotopia: Activation was present in nine studies, with involvement of the heterotopia or surrounding cortex in six, three of which had concomitant distant activation. Deactivation was also observed in nine studies, with involvement of the heterotopia and surrounding cortex in four, three of which had concomitant distant deactivation. Band heterotopia: Activation was present in all 11 studies, and always involved the heterotopia and surrounding cortex, 9 of which had concomitant distant activation. Deactivation was also observed in all 11 studies, with involvement of both the heterotopia and surrounding cortex, in addition to distant deactivation in 5 studies. EEG-fMRI studies reveal, non-invasively, metabolic responses in the heterotopia despite the fact that spikes are generated in the neocortex. The responses, activation or deactivation, had different correlation with the lesion and surrounding or distant cortex, activation reflecting intense neuronal activity, or excitation, and deactivation a possible distant (extra-lesional) inhibition. EEG-fMRI may become a useful tool to understand the epileptogenicity of such malformations. PMID- 16339794 TI - Interictal diffusion MRI in partial epilepsies explored with intracerebral electrodes. AB - Patients with refractory partial seizures may benefit from epilepsy surgery. However, invasive investigations are often needed to define the precise location and limits of the epileptogenic zone (EZ). In this study, we asked whether diffusion tensor imaging (DTI) might provide a non-invasive alternative to locate the EZ or at least provide insights to help place intracerebral electrodes for stereo-electroencephalography (SEEG). Whole brain DTI and voxel-based analysis (SPM99) was used to assess diffusion properties objectively in 16 epilepsy patients investigated with SEEG. Epilepsy was symptomatic in two patients and cryptogenic in the 14 remaining patients. The suspected onset of seizures was temporal in 10 patients, frontal in 2 and occipital in 4. Individual maps of apparent diffusion coefficient (ADC) and fractional anisotropy (FA) were calculated and compared to a database of 40 healthy volunteers. Thirteen of 16 patients exhibited diffusion abnormalities. ADC abnormalities were better correlated with SEEG data than FA abnormalities which were usually located at a distance or in the white matter. A significant increase in ADC (P < 0.01) was found in 11 patients and was located in the regions explored with depth electrodes in 7 of them. Surgery outcome was available in 3 of these 7 patients (2 were seizure free and 1 not). DTI specificity was better in extratemporal lobe epilepsy (83%) than in temporal lobe epilepsy (20%). When abnormalities concurred with the SEEG data, the concordance was optimal between the localization of the diffusion abnormalities and the irritative zone defined by SEEG. These encouraging, preliminary results, suggest that DTI examinations may provide accurate spatial data on the location and extent of the epileptogenic network in extratemporal lobe epilepsies. PMID- 16339795 TI - Neurotrophic factor synergy is required for neuronal survival and disinhibited axon regeneration after CNS injury. AB - The therapeutic effects of individual neurotrophic factors (NTF) have proved disappointing in clinical trials for neuronal repair and axon regeneration. Here, we demonstrate NTF synergistic neuronal responses after a combination of basic fibroblast growth factor, neurotrophin-3 and brain derived growth factor delivered to the somata of retinal ganglion cells promoted greater survival and axon growth than did the sum of the effects of each NTF alone. Triple and not single NTF treatments potentiated regulated intramembraneous proteolysis of p75(NTR), and ectodomain shedding of Nogo receptor, correlated with a 30% decrease in activation of Rho-A, a key signalling molecule in the axon growth inhibitory cascade. Thus, combinatorial NTF administration synergistically enhanced neuronal survival, disinhibited axon growth and promoted axon regeneration through the hostile CNS environment without the intervention of scar tissue at the lesion site. PMID- 16339796 TI - Global brain atrophy after unilateral parietal lesion and its prevention by erythropoietin. AB - In humans, neurotrauma is suspected to cause brain atrophy and accelerate slowly progressive neurodegenerative disorders, such as Alzheimer's disease or schizophrenia. However, a direct link between brain injury and subsequent delayed global neurodegeneration has remained elusive. Here we show that juvenile (4-week old) mice that are given a discrete unilateral lesion of the parietal cortex, develop to adulthood without obvious clinical symptoms. However, when monitored 3 and 9 months after lesioning, using high-resolution three-dimensional MRI and behavioural testing, the same mice display global neurodegenerative changes. Surprisingly, erythropoietin, a haematopoietic growth factor with potent neuroprotective activity, prevents behavioural abnormalities, cognitive dysfunction and brain atrophy when given for 2 weeks after acute brain injury. This demonstrates that a localized brain lesion is a primary cause of delayed global neurodegeneration that can be efficiently counteracted by neuroprotection. PMID- 16339797 TI - Intelligence and brain size in 100 postmortem brains: sex, lateralization and age factors. AB - The neural basis of variation in human intelligence is not well delineated. Numerous studies relating measures of brain size such as brain weight, head circumference, CT or MRI brain volume to different intelligence test measures, with variously defined samples of subjects have yielded inconsistent findings with correlations from approximately 0 to 0.6, with most correlations approximately 0.3 or 0.4. The study of intelligence in relation to postmortem cerebral volume is not available to date. We report the results of such a study on 100 cases (58 women and 42 men) having prospectively obtained Full Scale Wechsler Adult Intelligence Scale scores. Ability correlated with cerebral volume, but the relationship depended on the realm of intelligence studied, as well as the sex and hemispheric functional lateralization of the subject. General verbal ability was positively correlated with cerebral volume and each hemisphere's volume in women and in right-handed men accounting for 36% of the variation in verbal intelligence. There was no evidence of such a relationship in non-right-handed men, indicating that at least for verbal intelligence, functional asymmetry may be a relevant factor in structure-function relationships in men, but not in women. In women, general visuospatial ability was also positively correlated with cerebral volume, but less strongly, accounting for approximately 10% of the variance. In men, there was a non-significant trend of a negative correlation between visuospatial ability and cerebral volume, suggesting that the neural substrate of visuospatial ability may differ between the sexes. Analyses of additional research subjects used as test cases provided support for our regression models. In men, visuospatial ability and cerebral volume were strongly linked via the factor of chronological age, suggesting that the well documented decline in visuospatial intelligence with age is related, at least in right-handed men, to the decrease in cerebral volume with age. We found that cerebral volume decreased only minimally with age in women. This leaves unknown the neural substrate underlying the visuospatial decline with age in women. Body height was found to account for 1-4% of the variation in cerebral volume within each sex, leaving the basis of the well-documented sex difference in cerebral volume unaccounted for. With finer testing instruments of specific cognitive abilities and measures of their associated brain regions, it is likely that stronger structure-function relationships will be observed. Our results point to the need for responsibility in the consideration of the possible use of brain images as intelligence tests. PMID- 16339798 TI - Brain activation and hypothalamic functional connectivity during human non-rapid eye movement sleep: an EEG/fMRI study. AB - Regional differences in sleep EEG dynamics indicate that sleep-related brain activity involves local brain processes with sleep stage specific activity patterns of neuronal populations. Macroscopically, it is not fully understood which cerebral brain regions are involved in the successive discontinuation of wakefulness. We simultaneously used EEG and functional MRI on 9 subjects (6 female: mean = 24.1 years, 3 male: mean = 26.0 years) and analyzed local blood oxygenation level dependent signal changes linked to the transition from wakefulness to different non-rapid eye movement (NREM) sleep stages (according to Rechtschaffen and Kales) of the first sleep cycles after 36 h of total sleep deprivation. Several brain regions throughout the cortex, the limbic lobe, the thalamus, the caudate nucleus, as well as midbrain structures, such as the mammillary body/hypothalamus, showed reduced activity during NREM sleep across all sleep stages. Additionally, we found deactivation patterns specific to NREM sleep stages compared with wakefulness suggesting that a synchronized sleeping state can be established only if these regions interact in a well-balanced way. Sleep stage 2, which is usually linked to the loss of self-conscious awareness, is associated with signal decreases comprising thalamic and hypothalamic regions, the cingulate cortex, the right insula and adjacent regions of the temporal lobe, the inferior parietal lobule and the inferior/middle frontal gyri. The hypothalamic region known to be of particular importance in the regulation of the sleep-wake cycle shows specific temporally correlated network activity with the cortex while the system is in the sleeping state, but not during wakefulness. We describe a specific pattern of decreased brain activity during sleep and suggest that this pattern must be synchronized for establishing and maintaining sleep. PMID- 16339799 TI - The photorespiratory Arabidopsis shm1 mutant is deficient in SHM1. AB - Mitochondrial serine hydroxymethyltransferase (SHMT), combined with glycine decarboxylase, catalyzes an essential sequence of the photorespiratory C2 cycle, namely, the conversion of two molecules of glycine into one molecule each of CO2, NH4+, and serine. The Arabidopsis (Arabidopsis thaliana) mutant shm (now designated shm1-1) is defective in mitochondrial SHMT activity and displays a lethal photorespiratory phenotype when grown at ambient CO2, but is virtually unaffected at elevated CO2. The Arabidopsis genome harbors seven putative SHM genes, two of which (SHM1 and SHM2) feature predicted mitochondrial targeting signals. We have mapped shm1-1 to the position of the SHM1 gene (At4g37930). The mutation is due to a G --> A transition at the 5' splice site of intron 6 of SHM1, causing aberrant splicing and a premature termination of translation. A T DNA insertion allele of SHM1, shm1-2, and the F1 progeny of a genetic cross between shm1-1 and shm1-2 displayed the same conditional lethal phenotype as shm1 1. Expression of wild-type SHM1 under the control of either the cauliflower mosaic virus 35S or the SHM1 promoter in shm1-1 abrogated the photorespiratory phenotype of the shm mutant, whereas overexpression of SHM2 or expression of SHM1 under the control of the SHM2 promoter did not rescue the mutant phenotype. Promoter-beta-glucuronidase analyses revealed that SHM1 is predominantly expressed in leaves, whereas SHM2 is mainly transcribed in the shoot apical meristem and roots. Our findings establish SHM1 as the defective gene in the Arabidopsis shm1-1 mutant. PMID- 16339800 TI - ABA-hypersensitive germination3 encodes a protein phosphatase 2C (AtPP2CA) that strongly regulates abscisic acid signaling during germination among Arabidopsis protein phosphatase 2Cs. AB - The phytohormone abscisic acid (ABA) regulates physiologically important developmental processes and stress responses. Previously, we reported on Arabidopsis (Arabidopsis thaliana) L. Heynh. ahg mutants, which are hypersensitive to ABA during germination and early growth. Among them, ABA hypersensitive germination3 (ahg3) showed the strongest ABA hypersensitivity. In this study, we found that the AHG3 gene is identical to AtPP2CA, which encodes a protein phosphatase 2C (PP2C). Although AtPP2CA has been reported to be involved in the ABA response on the basis of results obtained by reverse-genetics approaches, its physiological relevance in the ABA response has not been clarified yet. We demonstrate in vitro and in vivo that the ahg3-1 missense mutation causes the loss of PP2C activity, providing concrete confirmation that this PP2C functions as a negative regulator in ABA signaling. Furthermore, we compared the effects of disruption mutations of eight structurally related PP2C genes of Arabidopsis, including ABI1, ABI2, HAB1, and HAB2, and found that the disruptant mutant of AHG3/AtPP2CA had the strongest ABA hypersensitivity during germination, but it did not display any significant phenotypes in adult plants. Northern-blot analysis clearly showed that AHG3/AtPP2CA is the most active among those PP2C genes in seeds. These results suggest that AHG3/AtPP2CA plays a major role among PP2Cs in the ABA response in seeds and that the functions of those PP2Cs overlap, but their unique tissue- or development-specific expression confers distinct and indispensable physiological functions in the ABA response. PMID- 16339803 TI - Putative Arabidopsis arp2/3 complex controls leaf cell morphogenesis. PMID- 16339801 TI - Heterotrimeric G proteins facilitate Arabidopsis resistance to necrotrophic pathogens and are involved in jasmonate signaling. AB - Heterotrimeric G proteins have been previously linked to plant defense; however a role for the Gbetagamma dimer in defense signaling has not been described to date. Using available Arabidopsis (Arabidopsis thaliana) mutants lacking functional Galpha or Gbeta subunits, we show that defense against the necrotrophic pathogens Alternaria brassicicola and Fusarium oxysporum is impaired in Gbeta-deficient mutants while Galpha-deficient mutants show slightly increased resistance compared to wild-type Columbia ecotype plants. In contrast, responses to virulent (DC3000) and avirulent (JL1065) strains of Pseudomonas syringae appear to be independent of heterotrimeric G proteins. The induction of a number of defense-related genes in Gbeta-deficient mutants were severely reduced in response to A. brassicicola infection. In addition, Gbeta-deficient mutants exhibit decreased sensitivity to a number of methyl jasmonate-induced responses such as induction of the plant defensin gene PDF1.2, inhibition of root elongation, seed germination, and growth of plants in sublethal concentrations of methyl jasmonate. In all cases, the behavior of the Galpha-deficient mutants is coherent with the classic heterotrimeric mechanism of action, indicating that jasmonic acid signaling is influenced by the Gbetagamma functional subunit but not by Galpha. We hypothesize that Gbetagamma acts as a direct or indirect enhancer of the jasmonate signaling pathway in plants. PMID- 16339804 TI - Nuclear actin-related proteins as epigenetic regulators of development. PMID- 16339805 TI - Use of two-color fluorescence-tagged transgenes to study interphase chromosomes in living plants. AB - Sixteen distinct sites distributed on all five Arabidopsis (Arabidopsis thaliana) chromosomes have been tagged using different fluorescent proteins and one of two different bacterial operator-repressor systems: (1) a yellow fluorescent protein Tet repressor fusion protein bound to tet operator sequences, or (2) a green or red fluorescent protein-Lac repressor fusion protein bound to lac operator sequences. Individual homozygous lines and progeny of intercrosses between lines have been used to study various aspects of interphase chromosome organization in root cells of living, untreated seedlings. Features reported here include distances between transgene alleles, distances between transgene inserts on different chromosomes, distances between transgene inserts on the same chromatin fiber, alignment of homologous chromosomes, and chromatin movement. The overall findings are consistent with a random and largely static arrangement of interphase chromosomes in nuclei of root cells. These transgenic lines provide tools for in-depth analyses of interphase chromosome organization, expression, and dynamics in living plants. PMID- 16339806 TI - Genome analysis and functional characterization of the E2 and RING-type E3 ligase ubiquitination enzymes of Arabidopsis. AB - Attachment of ubiquitin to substrate proteins is catalyzed by the three enzymes E1, E2 (ubiquitin conjugating [UBC]), and E3 (ubiquitin ligase). Forty-one functional proteins with a UBC domain and active-site cysteine are predicted in the Arabidopsis (Arabidopsis thaliana) genome, which includes four that are predicted or shown to function with ubiquitin-like proteins. Only nine were previously characterized biochemically as ubiquitin E2s. We obtained soluble protein for 22 of the 28 uncharacterized UBCs after expression in Escherichia coli and demonstrated that 16 function as ubiquitin E2s. Twelve, plus three previously characterized ubiquitin E2s, were also tested for the ability to catalyze ubiquitination in vitro in the presence of one of 65 really interesting new gene (RING) E3 ligases. UBC22, UBC19-20, and UBC1-6 had variable levels of E3 independent activity. Six UBCs were inactive with all RINGs tested. Closely related UBC8, 10, 11, and 28 were active with the largest number of RING E3s and with all RING types. Expression analysis was performed to determine whether E2s or E3s were expressed in specific organs or under specific environmental conditions. Closely related E2s show unique patterns of expression and most express ubiquitously. Some RING E3s are also ubiquitously expressed; however, others show organ-specific expression. Of all the organs tested, RING mRNAs are most abundant in floral organs. This study demonstrates that E2 diversity includes examples with broad and narrow specificity toward RINGs, and that most ubiquitin E2s are broadly expressed with each having a unique spatial and developmental pattern of expression. PMID- 16339807 TI - Structure and architecture of the maize genome. AB - Maize (Zea mays or corn) plays many varied and important roles in society. It is not only an important experimental model plant, but also a major livestock feed crop and a significant source of industrial products such as sweeteners and ethanol. In this study we report the systematic analysis of contiguous sequences of the maize genome. We selected 100 random regions averaging 144 kb in size, representing about 0.6% of the genome, and generated a high-quality dataset for sequence analysis. This sampling contains 330 annotated genes, 91% of which are supported by expressed sequence tag data from maize and other cereal species. Genes averaged 4 kb in size with five exons, although the largest was over 59 kb with 31 exons. Gene density varied over a wide range from 0.5 to 10.7 genes per 100 kb and genes did not appear to cluster significantly. The total repetitive element content we observed (66%) was slightly higher than previous whole-genome estimates (58%-63%) and consisted almost exclusively of retroelements. The vast majority of genes can be aligned to at least one sequence read derived from gene enrichment procedures, but only about 30% are fully covered. Our results indicate that much of the increase in genome size of maize relative to rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana) is attributable to an increase in number of both repetitive elements and genes. PMID- 16339808 TI - Major impact of admission glycaemia on 30 day and one year mortality in non diabetic patients admitted for myocardial infarction: results from the nationwide French USIC 2000 study. AB - OBJECTIVE: To analyse the short and long term prognostic significance of admission glycaemia in a large registry of non-diabetic patients with acute myocardial infarction. METHODS: Assessment of short and long term prognostic significance of admission blood glucose in a consecutive population of 1604 non diabetic patients admitted to intensive care units in France in November 2000 for a recent (5 h) decreases oxidative stress and augments K(+) currents, but not when excess ANG II is present. ANP has no effect on ventricular K(+) currents when the RAS is not activated, as in control males, in diabetic males treated with ACE inhibitor and in diabetic females. In conclusion, the modulation of K(+) currents and oxidative stress is significantly different in A and V cells in diabetic rat hearts. The evidence suggests that this is largely due to inhibition of RAS activation and/or action by ANP in A cells. These results may underlie chamber-specific arrhythmogenic mechanisms. PMID- 16339826 TI - Cardiac vagal modulation of heart rate during prolonged submaximal exercise in animals with healed myocardial infarctions: effects of training. AB - The present study investigated the effects of long-duration exercise on heart rate variability [as a marker of cardiac vagal tone (VT)]. Heart rate variability (time series analysis) was measured in mongrel dogs (n = 24) with healed myocardial infarctions during 1 h of submaximal exercise (treadmill running at 6.4 km/h at 10% grade). Long-duration exercise provoked a significant (ANOVA, all P < 0.01, means +/- SD) increase in heart rate (1st min, 165.3 +/- 15.6 vs. last min, 197.5 +/- 21.5 beats/min) and significant reductions in high frequency (0.24 to 1.04 Hz) power (VT: 1st min, 3.7 +/- 1.5 vs. last min, 1.0 +/- 0.9 ln ms(2)), R-R interval range (1st min, 107.9 +/- 38.3 vs. last min, 28.8 +/- 13.2 ms), and R-R interval SD (1st min, 24.3 +/- 7.7 vs. last min 6.3 +/- 1.7 ms). Because endurance exercise training can increase cardiac vagal regulation, the studies were repeated after either a 10-wk exercise training (n = 9) or a 10-wk sedentary period (n = 7). After training was completed, long-duration exercise elicited smaller increases in heart rate (pretraining: 1st min, 156.0 +/- 13.8 vs. last min, 189.6 +/- 21.9 beats/min; and posttraining: 1st min, 149.8 +/- 14.6 vs. last min, 172.7 +/- 8.8 beats/min) and smaller reductions in heart rate variability (e.g., VT, pretraining: 1st min, 4.2 +/- 1.7 vs. last min, 0.9 +/- 1.1 ln ms(2); and posttraining: 1st min, 4.8 +/- 1.1 vs. last min, 2.0 +/- 0.6 ln ms(2)). The response to long-duration exercise did not change in the sedentary animals. Thus the heart rate increase that accompanies long-duration exercise results, at least in part, from reductions in cardiac vagal regulation. Furthermore, exercise training attenuated these exercise-induced reductions in heart rate variability, suggesting maintenance of a higher cardiac vagal activity during exercise in the trained state. PMID- 16339827 TI - DNA sequence variation in the promoter region of the VEGF gene impacts VEGF gene expression and maximal oxygen consumption. AB - In its role as an endothelial cell proliferation and migration factor, vascular endothelial growth factor (VEGF) can affect peripheral circulation and therefore impact maximal oxygen consumption (Vo2 max). Because of the role of VEGF, and because variation in the VEGF gene has the ability to alter VEGF gene expression and VEGF protein level, we hypothesized that VEGF gene polymorphisms are related to VEGF gene expression in human myoblasts and Vo2 max before and after aerobic exercise training. We analyzed the effects of the VEGF -2578/-1154/-634 promoter region haplotype on VEGF gene expression by using a luciferase reporter assay in cultured human myoblasts and found that the AAG and CGC haplotypes resulted in significantly higher hypoxia-stimulated VEGF gene expression than the AGG and CGG haplotypes. Consistent with these results, we found that individuals with at least one copy of the AAG or CGC haplotype had higher Vo2 max before and after aerobic exercise training than did subjects with only the AGG and/or CGG haplotype. In conclusion, we found that VEGF -2578/-1154/-634 haplotype impacts VEGF gene expression in human myoblasts and is associated with Vo2 max. These results have potential implications for aerobic exercise training and may prove relevant in the study of pathological conditions that can be affected by angiogenesis, such as coronary artery disease and peripheral artery disease. PMID- 16339828 TI - Vasopressin V1 receptors contribute to hemodynamic and sympathoinhibitory responses evoked by stimulation of adenosine A2a receptors in NTS. AB - Activation of adenosine A2a receptors in the nucleus of the solitary tract (NTS) decreases mean arterial pressure (MAP), heart rate (HR), and renal sympathetic nerve activity (RSNA), whereas increases in preganglionic adrenal sympathetic nerve activity (pre-ASNA) occur, a pattern similar to that observed during hypotensive hemorrhage. Central vasopressin V1 receptors may contribute to posthemorrhagic hypotension and bradycardia. Both V1 and A2a receptors are densely expressed in the NTS, and both of these receptors are involved in cardiovascular control; thus they may interact. The responses elicited by NTS A2a receptors are mediated mostly via nonglutamatergic mechanisms, possibly via release of vasopressin. Therefore, we investigated whether blockade of NTS V1 receptors alters the autonomic response patterns evoked by stimulation of NTS A2a receptors (CGS-21680, 20 pmol/50 nl) in alpha-chloralose-urethane anesthetized male Sprague-Dawley rats. In addition, we compared the regional sympathetic responses to microinjections of vasopressin (0.1-100 ng/50 nl) into the NTS. Blockade of V1 receptors reversed the normal decreases in MAP into increases ( 95.6 +/- 28.3 vs. 51.4 +/- 15.7 integralDelta%), virtually abolished the decreases in HR (-258.3 +/- 54.0 vs. 18.9 +/- 57.8 integralDeltabeats/min) and RSNA (-239.3 +/- 47.4 vs. 15.9 +/- 36.1 integralDelta%), and did not affect the increases in pre-ASNA (279.7 +/- 48.3 vs. 233.1 +/- 54.1 integralDelta%) evoked by A2a receptor stimulation. The responses partially returned toward normal values approximately 90 min after the blockade. Microinjections of vasopressin into the NTS evoked dose-dependent decreases in HR and RSNA and variable MAP and pre-ASNA responses with a tendency toward increases. We conclude that the decreases in MAP, HR, and RSNA in response to NTS A2a receptor stimulation may be mediated via release of vasopressin from neural terminals in the NTS. The differential effects of NTS V1 and A2a receptors on RSNA versus pre-ASNA support the hypothesis that these receptor subtypes are differentially located/expressed on NTS neurons/neural terminals controlling different sympathetic outputs. PMID- 16339829 TI - Characterization of the role of gamma2 R531G mutation in AMP-activated protein kinase in cardiac hypertrophy and Wolff-Parkinson-White syndrome. AB - AMP-activated protein kinase (AMPK) is the downstream component of a protein kinase cascade that plays a key role in the regulation of energy metabolism. In humans, mutations in the gamma2-subunit of AMPK cause cardiac hypertrophy associated with Wolff-Parkinson-White syndrome, characterized by ventricular preexcitation. The effect of these mutations on AMPK activity and in development of the disease is enigmatic. Here we report that transgenic mice with cardiac specific expression of gamma2 harboring a mutation of arginine residue 531 to glycine (RG-TG) develop a striking cardiac phenotype by 4 wk of age, including hypertrophy, impaired contractile function, electrical conduction abnormalities, and marked glycogen accumulation. At this stage, AMPK activity isolated from hearts of RG-TG mice was almost completely abolished but could be restored after phosphorylation by an upstream AMPK kinase. At 1 wk of age, there was no detectable evidence of a cardiac phenotype, and AMPK activity in RG-TG hearts was similar to that in nontransgenic, control mice. We propose that mutations in gamma2 lead to suppression of total cardiac AMPK activity secondary to increased glycogen accumulation. The subsequent decrease in AMPK activity provides a mechanism that may explain the development of cardiac hypertrophy in this model. PMID- 16339830 TI - Effects of chronic activation of peroxisome proliferator-activated receptor-alpha or high-fat feeding in a rat infarct model of heart failure. AB - Intracardiac accumulation of lipid and related intermediates (e.g., ceramide) is associated with cardiac dysfunction and may contribute to the progression of heart failure (HF). Overexpression of nuclear receptor peroxisome proliferator activated receptor-alpha (PPARalpha) increases intramyocellular ceramide and left ventricular (LV) dysfunction. We tested the hypothesis that activation of fatty acid metabolism with fat feeding or a PPARalpha agonist increases myocardial triglyceride and/or ceramide and exacerbates LV dysfunction in HF. Rats with infarct-induced HF (n = 38) or sham-operated rats (n = 10) were either untreated (INF, n = 10), fed a high-fat diet (45% kcal fat, INF + Fat, n = 15), or fed the PPARalpha agonist fenofibrate (150 mg.kg(-1).day(-1), INF + Feno, n = 13) for 12 wk. LV ejection fraction was significantly reduced with HF (49 +/- 6%) compared with sham operated (86 +/- 2%) with no significant differences in ejection fraction (or other functional or hemodynamic measures) among the three infarcted groups. Treatment with the PPARalpha agonist resulted in LV hypertrophy (24% increase in LV/body mass ratio) and induced mRNAs encoding for PPARalpha regulated genes, as well as protein expression and activity of medium chain acyl CoA dehydrogenase (compared with INF and INF + Fat groups). Myocardial ceramide content was elevated in the INF group compared with sham-operated rats, with no further change in the INF + Fat or INF + Feno groups. Myocardial triglyceride was unaffected by infarction but increased in the INF + Fat group. In conclusion, LV dysfunction and dilation are not worsened despite upregulation of the fatty acid metabolic pathway and LV hypertrophy or accumulation of myocardial triglyceride in the rat infarct model of HF. PMID- 16339831 TI - Acidic extracellular pH-activated outwardly rectifying chloride current in mammalian cardiac myocytes. AB - Extracellular acidic pH was found to induce an outwardly rectifying Cl- current (I(Cl,acid)) in mouse ventricular cells, with a half-maximal activation at pH 5.9. The current showed the permeability sequence for anions to be SCN- > Br- > I > Cl- > F- > aspartate, while it exhibited a time-dependent activation at large positive potentials. Similar currents were also observed in mouse atrial cells and in atrial and ventricular cells from guinea pig. Some Cl- channel blockers (DIDS, niflumic acid, and glibenclamide) inhibited ICl,acid, whereas tamoxifen had little effect on it. Unlike volume-regulated Cl- current (ICl,vol) and CFTR Cl- current (ICl,CFTR), ICl,acid was independent of the presence of intracellular ATP. Activation of ICl,acid appeared to be also independent of intracellular Ca2+ and G protein. ICl,acid and ICl,vol could develop in an additive fashion in acidic hypotonic solutions. Isoprenaline-induced ICl,CFTR was inhibited by acidification in a pH-dependent manner in guinea pig ventricular cells. Our results support the view that ICl,acid and ICl,vol stem from two distinct populations of anion channels and that the ICl,acid channels are present in cardiac cells. ICl,acid may play a role in the control of action potential duration or cell volume under pathological conditions, such as ischemia-related cardiac acidosis. PMID- 16339832 TI - Thromboxane A2-induced arrhythmias in the anesthetized rabbit. AB - Experiments were conducted in the anesthetized rabbit to investigate mechanisms for arrhythmias that occur after left atrial injection of the thromboxane A(2) (TxA(2)) mimetic U-46619. Arrhythmias were primarily of ventricular origin, dose dependent in frequency, and TxA(2) receptor mediated. The response was receptor specific since arrhythmias were absent after pretreatment with a specific TxA(2) receptor antagonist (SQ-29548) and did not occur in response to another prostaglandin, PGF(2alpha). Alterations in coronary blood flow were unlikely the cause of these arrhythmias because coronary blood flow (as measured with fluorescent microspheres) was unchanged after U-46619, and there were no observable changes in the ECG-ST segment. In addition, arrhythmias did not occur after administration of another vasoconstrictor (phenylephrine). The potential involvement of autonomic cardiac efferent nerves in these arrhythmias was also investigated because TxA(2) has been shown to stimulate peripheral nerves. Pretreatment of animals with the beta-adrenergic receptor antagonist propranolol did not reduce the frequency of these arrhythmias. Pretreatment with atropine or bilateral vagotomy resulted in an increased frequency of arrhythmias, suggesting that parasympathetic nerves may actually inhibit the arrhythmogenic activity of TxA(2). These experiments demonstrate that left atrial injection of U-46619 elicits arrhythmias via a mechanism independent of a significant reduction in coronary blood flow or activation of the autonomic nervous system. It is possible that TxA(2) may have a direct effect on the electrical activity of the heart in vivo, which provides significant implications for cardiac events where TxA(2) is increased, e.g., after myocardial ischemia or administration of cyclooxygenase-2 inhibitors. PMID- 16339833 TI - Cerebrovascular response to normal pregnancy: a longitudinal study. AB - We used a longitudinal study design (gestational weeks 8, 15, 22, 29, and 36 and 12 wk postpartum ) to investigate the effect of normal pregnancy on cerebral autoregulation and pressor response. Blood flow velocities in the right internal carotid artery, end-tidal CO2, and mean arterial pressure (MAP) were simultaneously and continuously recorded in 16 healthy pregnant women during standardized hyperventilation and handgrip. Blood flow velocities were recorded using Doppler ultrasound sampled beat by beat using the ECG signal. The results demonstrate that the vasoconstrictor response to hyperventilation is unchanged during pregnancy. During standardized handgrip, MAP showed a statistically significant increase during pregnancy that did not affect cerebral blood flow. A statistically significant reduction in the MAP response to handgrip was seen in week 36. In conclusion, pregnancy has no impact on cerebral autoregulation. There is an impact on the pressor response resulting in a blunted reaction at week 36, probably caused by a fall in the baroreflex set point. PMID- 16339834 TI - Duration of no-load state affects opening angle of porcine coronary arteries. AB - The zero-stress state of a blood vessel has been extensively studied because it is the reference state for which all calculations of intramural stress and strain must be based. It has also been found to reflect nonuniformity in growth and remodeling in response to chemical or physical changes. The zero-stress state can be characterized by an opening angle, defined as the angle subtended by two radii connecting the midpoint of the inner wall. All prior studies documented the zero stress state or opening angle with no regard to duration of the no-load state. Our hypotheses were that, given the viscoelastic properties of blood vessels, the zero-stress state may have "memory" of prior circumferential and axial loading, i.e., duration of the no-load state influences opening angle. To test these hypotheses, we considered ring pairs of porcine coronary arteries to examine the effect of duration in the no-load state after circumferential distension. Our results show a significant reduction in opening angle as duration of the no-load state increases, i.e., vessels that are reduced to the zero-stress state directly from the loaded state attain much larger opening angles at 30 min after the radial cut than rings that are in the no-load state for various durations. To examine the effect of axial loading, we found similar reductions in opening angle with duration in the no-load from the in situ state, albeit the effect was significantly smaller than that of circumferential loading. Hence, we found that the zero-stress state has memory of both circumferential and axial loading. These results are important for understanding viscoelastic properties of coronary arteries, interpretation of the enormous data on the opening angle and strain in the literature, and standardization of future measurements on the zero-stress state. PMID- 16339835 TI - Nitric oxide-cGMP-protein kinase G signaling pathway induces anoxic preconditioning through activation of ATP-sensitive K+ channels in rat hearts. AB - Nitric oxide (NO) plays an important role in anoxic preconditioning to protect the heart against ischemia-reperfusion injuries. The present work was performed to study better the NO-cGMP-protein kinase G (PKG) signaling pathway in the activation of both sarcolemmal and mitochondrial ATP-sensitive K+ (KATP) channels during anoxic preconditioning (APC) and final influence on reducing anoxia reperfusion (A/R)-induced cardiac damage in rat hearts. The upstream regulating elements controlling NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection were investigated. The involvement of both inducible and endothelial NO synthases (iNOS and eNOS) in the progression of this signaling pathway was followed. Final cellular outcomes of ischemia-induced injury after different preconditioning in the form of lactate dehydrogenase release, DNA strand breaks, and malondialdehyde formation as indexes of cell injury and lipid peroxidation, respectively, were investigated. The lactate dehydrogenase and malondialdehyde values decreased in the groups that underwent preconditioning periods with specific mitochondrial KATP channels opener diazoxide (100 microM), nonspecific mitochondrial KATP channels opener pinacidil (50 microM), S-nitroso-N acetylpenicillamine (SNAP, 300 microM), or beta-phenyl-1,N2-etheno-8 bromoguanosine-3',5'-cyclicmonophosphorothioate, Sp-isomer (10 microM) before the A/R period. Preconditioning with SNAP significantly reduced the DNA damage. The effect was blocked by glibenclamide (50 microM), 5-hydroxydecanoate (100 microM), NG-nitro-L-arginine methyl ester (200 microM), and beta-phenyl-1,N2-etheno-8 bromoguanosine-3',5'-cyclic monophosphorothioate, Rp-isomer (1 microM). The results suggest iNOS, rather than eNOS, as the major contributing NO synthase during APC treatment. Moreover, the PKG shows priority over NO as the upstream regulator of NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection during APC treatment. PMID- 16339836 TI - Synergistic targeting with bone marrow-derived cells and PDGF improves diabetic vascular function. AB - Diabetes mellitus is associated with an increased risk of vascular disease, with significant alterations in systemic endothelial progenitor cells (EPCs) and peripheral vascular function. To identify the contribution of the different vascular compartments in the diabetic impairment of vascularization, we employed streptozotocin- and control-treated 3-mo-old C57Bl/6 mice in an isogeneic pinnal cardiac allograft model, revealing a significant delay in vascularization of wild type cardiac tissue transplanted into diabetic mice. To investigate the basis of this impairment, the function of diabetic bone marrow cells was tested by transplantation of bone marrow cells isolated from diabetic and control mice into intact, unirradiated 18-mo-old C57Bl/6 mice, which have impaired function of both EPCs and peripheral endothelial cells. Importantly, cells derived from control, but not diabetic, bone marrow integrated into transplanted cardiac allografts. To assess the contribution of diabetic changes in the local vasculature, diabetic mice were treated with pinnal injections of platelet-derived growth factor (PDGF) AB, which promotes cardiac angiogenesis in wild-type mice. However, whereas PDGF AB enhanced allograft function in control mice, the activity of the cardiac transplants in the PDGF-AB-treated diabetic mice was significantly decreased. To decipher the potential interactions between systemic bone marrow-derived cells and local vascular pathways, diabetic mice were transplanted with wild-type bone marrow cells with or without PDGF-AB pinnal pretreatment, resulting in improved allograft function and donor cell recruitment only in the combination treatment arm. Overall, these studies show that the diabetic impairment in cardiac angiogenesis can be reversed by targeting the synergism between local trophic pathways and systemic cell function. PMID- 16339837 TI - Activation of Nrf2/ARE pathway protects endothelial cells from oxidant injury and inhibits inflammatory gene expression. AB - The antioxidant response element (ARE) is a transcriptional control element that mediates expression of a set of antioxidant proteins. NF-E2-related factor 2 (Nrf2) is a transcription factor that activates ARE-containing genes. In endothelial cells, the ARE-mediated genes are upregulated by atheroprotective laminar flow through a Nrf2-dependent mechanism. We tested the hypothesis that activation of ARE-regulated genes via adenovirus-mediated expression of Nrf2 may suppress redox-sensitive inflammatory gene expression. Expression of Nrf2 in human aortic endothelial cells (HAECs) resulted in a marked increase in ARE driven transcriptional activity and protected HAECs from H2O2-mediated cytotoxicity. Nrf2 suppressed TNF-alpha-induced monocyte chemoattractant protein (MCP)-1 and VCAM-1 mRNA and protein expression in a dose-dependent manner and inhibited TNF-alpha-induced monocytic U937 cell adhesion to HAECs. Nrf2 also inhibited IL-1beta-induced MCP-1 gene expression in human mesangial cells. Expression of Nrf2 inhibited TNF-alpha-induced activation of p38 MAP kinase. Furthermore, expression of a constitutively active form of MKK6 (an upstream kinase for p38 MAP kinase) partially reversed Nrf2-mediated inhibition of VCAM-1 expression, suggesting that p38 MAP kinase, at least in part, mediates Nrf2's anti-inflammatory action. In contrast, Nrf2 did not inhibit TNF-alpha-induced NF kappaB activation. These data identify the Nrf2/ARE pathway as an endogenous atheroprotective system for antioxidant protection and suppression of redox sensitive inflammatory genes, suggesting that targeting the Nrf2/ARE pathway may represent a novel therapeutic approach for the treatment of inflammatory diseases such as atherosclerosis. PMID- 16339838 TI - Effects of chronic portal hypertension on agonist-induced actin polymerization in small mesenteric arteries. AB - The ability of arterial smooth muscle to respond to vasoconstrictor stimuli is reduced in chronic portal hypertension (PHT). Additional evidence supports the existence of a postreceptor defect in vascular smooth muscle excitation contraction coupling. However, the nature of this defect is unclear. Recent studies have shown that vasoconstrictor stimuli induce actin polymerization in smooth muscle and that the associated increase in F-actin is necessary for force development. In the present study we have tested the hypothesis that impaired actin polymerization contributes to reduced vasoconstrictor function in small mesenteric arteries derived from rats with chronic prehepatic PHT. In vitro studies were conducted on small mesenteric artery vessel rings isolated from normal and PHT rats. Isometric tension responses to incremental concentrations of phenylephrine were significantly reduced in PHT arteries. The ability to polymerize actin in portal hypertensive mesenteric arteries stimulated by phenylephrine was attenuated compared with control. Inhibition of cAMP-dependent protein kinase (PKA) restored agonist-induced actin polymerization of arteries from PHT rats to normal levels. Depolymerization of actin in arteries from normal rats reduced maximal contractile force but not myosin phosphorylation, suggesting a key role for the dynamic regulation of actin polymerization in the maintenance of vascular smooth muscle contraction. We conclude that reductions in agonist induced maximal force development of PHT vascular smooth muscle is due, in part, to impaired actin polymerization, and prolonged PKA activation may underlie these changes. PMID- 16339839 TI - The PPAR-alpha activator fenofibrate fails to provide myocardial protection in ischemia and reperfusion in pigs. AB - Rodent studies suggest that peroxisome proliferator-activated receptor-alpha (PPAR-alpha) activation reduces myocardial ischemia-reperfusion (I/R) injury and infarct size; however, effects of PPAR-alpha activation in large animal models of myocardial I/R are unknown. We determined whether chronic treatment with the PPAR alpha activator fenofibrate affects myocardial I/R injury in pigs. Domestic farm pigs were assigned to treatment with fenofibrate 50 mg.kg(-1).day(-1) orally or no drug treatment, and either a low-fat (4% by weight) or a high-fat (20% by weight) diet. After 4 wk, 66 pigs underwent 90 min low-flow regional myocardial ischemia and 120 min reperfusion under anesthetized open-chest conditions, resulting in myocardial stunning. The high-fat group received an infusion of triglyceride emulsion and heparin during this terminal experiment to maintain elevated arterial free fatty acid (FFA) levels. An additional 21 pigs underwent 60 min no-flow ischemia and 180 min reperfusion, resulting in myocardial infarction. Plasma concentration of fenofibric acid was similar to the EC50 for activation of PPAR-alpha in vitro and to maximal concentrations achieved in clinical use. Myocardial expression of PPAR-alpha mRNA was prominent but unaffected by fenofibrate treatment. Fenofibrate increased expression of carnitine palmitoyltransferase (CPT)-I mRNA in liver and decreased arterial FFA and lactate concentrations (each P < 0.01). However, fenofibrate did not affect myocardial CPT-I expression, substrate uptake, lipid accumulation, or contractile function during low-flow I/R in either the low- or high-fat group, nor did it affect myocardial infarct size. Despite expression of PPAR-alpha in porcine myocardium and effects of fenofibrate on systemic metabolism, treatment with this PPAR-alpha activator does not alter myocardial metabolic or contractile responses to I/R in pigs. PMID- 16339840 TI - ANG II induces apoptosis of human vascular smooth muscle via extrinsic pathway involving inhibition of Akt phosphorylation and increased FasL expression. AB - In addition to well-documented vascular growth-promoting effects, ANG II exerts proapoptotic effects that are poorly understood. IGF-1 is a potent survival factor for human vascular smooth muscle cells (hVSMC), and its antiapoptotic effects are mediated via the IGF-1 receptor (IGF-1R) through a signaling pathway involving phosphatidylinositol 3-kinase and Akt. We hypothesized that there would be cross talk between ANG II proapoptotic effects and IGF-1 survival effects in hVSMC. To investigate ANG II-induced apoptosis and the potential involvement of IGF-1, we exposed quiescent and nonquiescent hVSMC to ANG II. ANG II induced apoptosis only in nonquiescent cells but stimulated hypertrophy in quiescent cells. ANG II-induced apoptosis was characterized by marked inhibition of Akt phosphorylation and stimulation of membrane Fas ligand (FasL) expression, caspase 8 activation, and a reduction in soluble FasL expression. Adenovirally mediated overexpression of Akt rescued hVSMC from ANG II-induced apoptosis. IGF-1R activation increased Akt phosphorylation and soluble FasL expression, and these effects were completely blocked by coincubating hVSMC with ANG II. In conclusion, ANG II-induced apoptosis of hVSMC is characterized by marked inhibition of Akt phosphorylation and stimulation of an extrinsic cell death signaling pathway via upregulation of membrane FasL expression, caspase-8 activation, and a reduction in soluble FasL expression. Furthermore, ANG II antagonizes the antiapoptotic effect of IGF-1 by blocking its ability to increase Akt phosphorylation and soluble FasL. These findings provide novel insights into ANG II-induced apoptotic signaling and have significant implication for understanding ANG II-induced remodeling in hypertension and atherosclerosis. PMID- 16339841 TI - Bioenergetic protection of failing atrial and ventricular myocardium by vasopeptidase inhibitor omapatrilat. AB - Deficient bioenergetic signaling contributes to myocardial dysfunction and electrical instability in both atrial and ventricular cardiac chambers. Yet, approaches capable to prevent metabolic distress are only partially established. Here, in a canine model of tachycardia-induced congestive heart failure, we compared atrial and ventricular bioenergetics and tested the efficacy of metabolic rescue with the vasopeptidase inhibitor omapatrilat. Despite intrinsic differences in energy metabolism, failing atria and ventricles demonstrated profound bioenergetic deficiency with reduced ATP and creatine phosphate levels and compromised adenylate kinase and creatine kinase catalysis. Depressed phosphotransfer enzyme activities correlated with reduced tissue ATP levels, whereas creatine phosphate inversely related with atrial and ventricular load. Chronic treatment with omapatrilat maintained myocardial ATP, the high-energy currency, and protected adenylate and creatine kinase phosphotransfer capacity. Omapatrilat-induced bioenergetic protection was associated with maintained atrial and ventricular structural integrity, albeit without full recovery of the creatine phosphate pool. Thus therapy with omapatrilat demonstrates the benefit in protecting phosphotransfer enzyme activities and in preventing impairment of atrial and ventricular bioenergetics in heart failure. PMID- 16339842 TI - Electrical remodeling of cardiac myocytes from mice with heart failure due to the overexpression of tumor necrosis factor-alpha. AB - Mice that overexpress the inflammatory cytokine tumor necrosis factor-alpha in the heart (TNF mice) develop heart failure characterized by atrial and ventricular dilatation, decreased ejection fraction, atrial and ventricular arrhythmias, and increased mortality (males > females). Abnormalities in Ca2+ handling, prolonged action potential duration (APD), calcium alternans, and reentrant atrial and ventricular arrhythmias were previously observed with the use of optical mapping of perfused hearts from TNF mice. We therefore tested whether altered voltage-gated outward K+ and/or inward Ca2+ currents contribute to the altered action potential characteristics and the increased vulnerability to arrhythmias. Whole cell voltage-clamp recordings of K+ currents from left ventricular myocytes of TNF mice revealed an approximately 50% decrease in the rapidly activating, rapidly inactivating transient outward K+ current Ito and in the rapidly activating, slowly inactivating delayed rectifier current IK,slow1, an approximately 25% decrease in the rapidly activating, slowly inactivating delayed rectifier current IK,slow2, and no significant change in the steady-state current Iss compared with controls. Peak amplitudes and inactivation kinetics of the L-type Ca2+ current ICa,L were not altered. Western blot analyses revealed a reduction in the proteins underlying Kv4.2, Kv4.3, and Kv1.5. Thus decreased K+ channel expression is largely responsible for the prolonged APD in the TNF mice and may, along with abnormalities in Ca2+ handling, contribute to arrhythmias. PMID- 16339843 TI - Molecular characterization of methicillin-resistant Staphylococcus aureus bloodstream isolates from Croatia. AB - OBJECTIVES: The objectives of this study were (i) to investigate the genetic background of methicillin-resistant Staphylococcus aureus (MRSA) bloodstream isolates from Croatia and (ii) to monitor the prevalence of Panton-Valentine leucocidin (PVL) and toxic shock syndrome toxin-1 (TSST-1) among these isolates. METHODS: Eighty-two hospital-acquired MRSA bloodstream isolates, collected in 2001 and 2002 in Croatia, were characterized by PFGE, staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). The presence of genes encoding PVL and TSST-1 was investigated by real-time PCR. RESULTS: All strains were multiresistant and were distributed among 16 different similarity groups as determined by PFGE. Two of the groups, groups H and K, harboured the majority of the MRSA strains with 52 and 12%, respectively. The predominant SCCmec type found among the isolates was type I (89%). Eleven per cent of the strains harboured a modified SCCmec type III, which contained, in contrast to the regular type III, an additional dcs region. One strain harboured a novel SCCmec type, containing the ccrC gene in combination with the mecI gene, the dcs region, the locus between pI258 and Tn554 (locus E) and the locus between Tn554 and orfX (locus F). MLST showed the presence of ST111-MRSA-I and ST247-MRSA I among Croatian MRSA isolates. All isolates were negative for both PVL and TSST 1. CONCLUSIONS: These results indicate the emergence of ST111-MRSA-I and ST247 MRSA-I in Croatia among MRSA bloodstream isolates. The virulence factors PVL and TSST-1 were not present among these isolates. PMID- 16339844 TI - Effect of triclosan on the development of bacterial biofilms by urinary tract pathogens on urinary catheters. AB - OBJECTIVES: To examine (i) the effect of triclosan on the formation of catheter biofilms by urinary tract pathogens and (ii) the diffusion of triclosan through the retention balloons of urinary catheters. METHODS: Models of the catheterized bladder were infected with eight different urinary tract pathogens and the effect of triclosan on biofilm formation was assessed by determining the numbers of viable cells colonizing the catheters and by scanning electron microscopy. HPLC was used to determine the triclosan concentration in urine draining from models that had been fitted with triclosan-inflated silicone catheters. RESULTS: When catheters were inflated with triclosan (10 g/L) the formation of catheter biofilm by Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Proteus mirabilis was prevented. The numbers of Enterococcus faecalis and Providencia stuartii cells colonizing catheters were also significantly reduced (P<0.05). Serratia marcescens, Morganella morganii and Pseudomonas aeruginosa, however, were able to produce extensive catheter biofilms in the presence of triclosan. Only P. mirabilis produced alkaline urine and encrusted the catheters. Concentrations of 0.02-0.16 mg/L of the biocide were detected in urine draining from the model over the 48 h experimental period. CONCLUSIONS: Triclosan diffused through silicone catheter balloons and produced urinary concentrations that prevented catheter encrustation by P. mirabilis and biofilm formation by several other common pathogens of the catheterized urinary tract. It had little effect on urease-producing P. aeruginosa, S. marcescens or M. morganii but these species did not produce alkaline urine or crystalline biofilms. PMID- 16339845 TI - Evidence for a multivalent interaction of symmetrical, N-linked, lidocaine dimers with voltage-gated Na+ channels. AB - The interaction of symmetrical lidocaine dimers with voltage-gated Na+ channels (VGSCs) was examined using a FLIPR membrane potential assay and voltage-clamp. The dimers, in which the tertiary amines of the lidocaine moieties are linked by an alkylene chain (two to six methylene units), inhibited VGSC activator-evoked depolarization of cells heterologously-expressing rat (r) Na(v)1.2a, human (h) Na(v)1.5, and rNa(v)1.8, with potencies 10- to 100-fold higher than lidocaine (compound 1). The rank order of potency (C4 (compound 4) > C3 (compound 3) > or = C2 (compound 2) = C5 (compound 5) = C6 (compound 6) >> compound 1) was similar at each VGSC. Compound 4 exhibited strong use-dependent inhibition of hNa(v)1.5 with pIC50 values < 4.5 and 6.0 for tonic and phasic block, respectively. Coincubation with local anesthetics but not tetrodotoxin attenuated compound 4-mediated inhibition of hNa(v)1.5. These data suggest that the compound 4 binding site(s) is identical, or allosterically coupled, to the local anesthetic receptor. The dissociation rate of the dimers from hNa(v)1.5 was dependent upon the linker length, with a rank order of compound 1 > compound 5 = compound 6 > compound 2 >> compound 3. The observation that both the potency and dissociation rate of the dimers was dependent upon linker length is consistent with a multivalent interaction at VGSCs. hNa(v)1.5 VGSCs did not recover from inhibition by compound 4. However, "chase" with free local anesthetic site inhibitors increased the rate of dissociation of compound 4. Together, these data support the hypothesis that compound 4 simultaneously occupies two binding sites on VGSCs, both of which can be bound by known local anesthetic site inhibitors. PMID- 16339846 TI - Novel features of G protein-coupled receptor kinase 4. AB - The defining characteristic of G protein-coupled receptor homologous desensitization is that the receptor must be occupied by an agonist or in an activated conformation that mimics an agonist-induced state. In most instances, the mechanistic basis for this characteristic is the high selectivity of G protein-coupled receptor kinases for the activated receptor. In this issue, Rankin et al. (p. 759) demonstrate that under some conditions, at least, the G protein-coupled receptor kinase GRK4 does not display a preference for the agonist-occupied D1 dopamine receptor. Coexpression of GRK4 and the D1 receptor in a heterologous system induces phosphorylation of the receptor in the absence of agonist, causing constitutive desensitization and internalization of the receptor. Lacking the normal rapid feedback mechanisms associated with homologous desensitization, a system incorporating constitutively active GRK4 will be prone to dysregulation, perhaps explaining the generally low expression of GRK4. Indeed, considerable evidence suggests that just such dysregulation resulting from mutationally activated GRK4 contributes to the heritable component of human essential hypertension (Physiol Genomics 19:223-246, 2004). PMID- 16339847 TI - The ubiquitin-specific protease Usp4 regulates the cell surface level of the A2A receptor. AB - Many membrane proteins incur a folding problem during biosynthesis; only a fraction thereof is exported from the endoplasmic reticulum (ER), because quality control is stringent. This is also true for G protein-coupled receptors. Here, we identify the deubiquitinating enzyme Usp4 as an interaction partner of the A2a adenosine receptor, a Gs-coupled receptor. Usp4 binds to the carboxyl terminus of the A2A receptor and allows for its accumulation as deubiquinated protein. This relaxes ER quality control and enhances cell surface expression of functionally active receptor. The effect of Usp4 on the A2A receptor was specific because 1) it was not seen in C-terminally truncated versions of the receptor; 2) it was not mimicked by Usp14, another member of the ubiquitin-specific protease family; and 3) it was not seen with the metabotropic glutamate receptor-5, another G protein coupled receptor with a high propensity for intracellular retention. These observations show that deubiquinating enzymes can regulate quality control in the ER. PMID- 16339848 TI - Advances in genetics: what are the benefits for patients? PMID- 16339849 TI - Evidence for susceptibility determinant(s) to psoriasis vulgaris in or near PTPN22 in German patients. AB - INTRODUCTION: Variant R620W of protein tyrosine phosphatase non-receptor type 22 (PTPN22) has consistently been reported as a susceptibility factor for several autoimmune diseases. We investigated its role in susceptibility to psoriasis, the relevance of possibly other disease-causing variants, and interdependency of the major risk factor for psoriasis at PSORS1. METHODS: R620W was tested in a case control study initially with 375 German patients and then with an enlarged sample of an additional 418 patients. Analyses were extended to linkage disequilibrium (LD) based haplotypes. Potential interaction between risk haplotypes of PTPN22 and the PSORS1 associated risk allele was tested by regression analysis. PTPN22 coding sequence was determined in 20 patients carrying the risk haplotype. Association and regression analysis were also performed in the extended case control study. RESULTS: R620W was not associated in either case-control study, while significant association (corrected for multiple testing) with one haplotype (C-4) of the LD block encompassing PTPN22 as well with another haplotype (B-3) within an adjacent telomeric LD block was detected. No evidence for interaction between risk haplotype C-4 and the PSORS1 associated risk allele was found. Sequencing excluded other coding variants within PTPN22 as a basis for association findings. Analysis of the extended study group confirmed association for haplotypes B-3 and C-4 and independence of risk haplotypes C-4 and PSORS1. DISCUSSION: We exclude a major role of *620W in German psoriasis patients but suggest that other susceptibility determinant(s) within non-coding regions of PTPN22 or its proximity might exist acting independently of the major PSORS1 risk factor. PMID- 16339850 TI - Arabidopsis CONSTANS-LIKE3 is a positive regulator of red light signaling and root growth. AB - CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) is an E3 ubiquitin ligase that represses photomorphogenesis in the dark. Therefore, proteins interacting with COP1 could be important regulators of light-dependent development. Here, we identify CONSTANS-LIKE3 (COL3) as a novel interaction partner of COP1. A green fluorescent protein-COL3 fusion protein colocalizes with COP1 to nuclear speckles when transiently expressed in plant cells. This localization requires the B-box domains in COL3, indicating a novel function of this domain. A loss-of-function col3 mutant has longer hypocotyls in red light and in short days. Unlike constans, the col3 mutant flowers early and shows a reduced number of lateral branches in short days. The mutant also exhibits reduced formation of lateral roots. The col3 mutation partially suppresses the cop1 and deetiolated1 (det1) mutations in the dark, suggesting that COL3 acts downstream of both of these repressors. However, the col3 mutation exerts opposing effects on cop1 and det1 in terms of lateral roots and anthocyanin accumulation, suggesting that COL3 also has activities that are independent of COP1 and DET1. In conclusion, we have identified COL3 as a positive regulator of photomorphogenesis that acts downstream of COP1 but can promote lateral root development independently of COP1 and also function as a daylength-sensitive regulator of shoot branching. PMID- 16339851 TI - Chloroplast biogenesis of photosystem II cores involves a series of assembly controlled steps that regulate translation. AB - The biogenesis of photosystem II, one of the major photosynthetic protein complexes, involves a cascade of assembly-governed regulation of translation of its major chloroplast-encoded subunits. In Chlamydomonas reinhardtii, the presence of the reaction center subunit D2 is required for the expression of the other reaction center subunit D1, while the presence of D1 is required for the expression of the core antenna subunit apoCP47. Using chimeric genes expressed in the chloroplast, we demonstrate that the decreased synthesis of D1 or apoCP47 in the absence of protein assembly is due to a genuine downregulation of translation. This regulation is mediated by the 5' untranslated region of the corresponding mRNA and originates from negative feedback exerted by the unassembled D1 or apoCP47 polypeptide. However, autoregulation of translation of subunit D1 is not implicated in the recovery from photoinhibition, which involves an increased translation of psbA mRNA in response to the degradation of photodamaged D1. De novo synthesis and repair of photosystem II complexes are independently controlled. PMID- 16339852 TI - The serine/arginine-rich protein family in rice plays important roles in constitutive and alternative splicing of pre-mRNA. AB - Ser/Arg-rich (SR) proteins play important roles in the constitutive and alternative splicing of pre-mRNA. We isolated 20 rice (Oryza sativa) genes encoding SR proteins, of which six contain plant-specific characteristics. To determine whether SR proteins modulate splicing efficiency and alternative splicing of pre-mRNA in rice, we used transient assays in rice protoplasts by cotransformation of SR protein genes with the rice Waxy(b) (Wx(b))-beta glucuronidase fusion gene. The results showed that plant-specific RSp29 and RSZp23, an SR protein homologous to human 9G8, enhanced splicing and altered the alternative 5' splice sites of Wx(b) intron 1. The resulting splicing pattern was unique to each SR protein; RSp29 stimulated splicing at the distal site, and RSZp23 enhanced splicing at the proximal site. Results of domain-swapping experiments between plant-specific RSp29 and SCL26, which is a homolog of human SC35, showed the importance of RNA recognition motif 1 and the Arg/Ser-rich (RS) domain for the enhancement of splicing efficiencies. Overexpression of plant specific RSZ36 and SRp33b, a homolog of human ASF/SF2, in transgenic rice changed the alternative splicing patterns of their own pre-mRNAs and those of other SR proteins. These results show that SR proteins play important roles in constitutive and alternative splicing of rice pre-mRNA. PMID- 16339853 TI - A new role for the Arabidopsis AP2 transcription factor, LEAFY PETIOLE, in gibberellin-induced germination is revealed by the misexpression of a homologous gene, SOB2/DRN-LIKE. AB - Gibberellic acid (GA) promotes germination, stem/hypocotyl elongation, and leaf expansion during seedling development. Using activation-tagging mutagenesis, we identified a mutation, sob2-D (for suppressor of phytochromeB-4 [phyB-4]#2 dominant), which suppresses the long-hypocotyl phenotype of a phyB missense allele, phyB-4. This mutant phenotype is caused by the overexpression of an APETALA2 transcription factor, SOB2, also called DRN-like. SOB2/DRN-like transcript is not detectable in wild-type seedling or adult tissues via RT-PCR analysis, suggesting that SOB2/DRN-like may not be involved in seedling development under normal conditions. Adult sob2-D phyB-4 plants have curled leaves and club-like siliques, resembling plants that overexpress a closely related gene, LEAFY PETIOLE (LEP). Hypocotyls of a LEP-null allele, lep-1, are shorter in the light and dark, suggesting LEP involvement in seedling development. This aberrant hypocotyl phenotype is due at least in part to a delay in germination. In addition, lep-1 is less responsive to GA and more sensitive to the GA biosynthesis inhibitor paclobutrazol, indicating that LEP is a positive regulator of GA-induced germination. RT-PCR shows that LEP transcript accumulates in wild-type seeds during imbibition and germination, and the transcript levels of REPRESSOR OF ga1-3-LIKE2 (RGL2), a negative regulator of GA signaling during germination, is unaffected in lep-1. These results suggest LEP is a positive regulator of GA-induced germination acting independently of RGL2. An alternative model places LEP downstream of RGL2 in the GA-signaling cascade. PMID- 16339854 TI - Golgi-mediated vacuolar sorting of the endoplasmic reticulum chaperone BiP may play an active role in quality control within the secretory pathway. AB - Quality control in the endoplasmic reticulum (ER) prevents the arrival of incorrectly or incompletely folded proteins at their final destinations and targets permanently misfolded proteins for degradation. Such proteins have a high affinity for the ER chaperone BiP and are finally degraded via retrograde translocation from the ER lumen back to the cytosol. This ER-associated protein degradation (ERAD) is currently thought to constitute the main disposal route, but there is growing evidence for a vacuolar role in quality control. We show that BiP is transported to the vacuole in a wortmannin-sensitive manner in tobacco (Nicotiana tabacum) and that it could play an active role in this second disposal route. ER export of BiP occurs via COPII-dependent transport to the Golgi apparatus, where it competes with other HDEL receptor ligands. When HDEL mediated retrieval from the Golgi fails, BiP is transported to the lytic vacuole via multivesicular bodies, which represent the plant prevacuolar compartment. We also demonstrate that a subset of BiP-ligand complexes is destined to the vacuole and differs from those likely to be disposed of via the ERAD pathway. Vacuolar disposal could act in addition to ERAD to maximize the efficiency of quality control in the secretory pathway. PMID- 16339856 TI - Cross-kingdom hormonal signaling: an insight from thyroid hormone functions in marine larvae. AB - Thyroid hormones (THs) are small, lipophilic signaling molecules built from tyrosine and iodine. TH action is well characterized in vertebrates, where these molecules play a fundamental role as regulators of development, metabolism, growth and differentiation. Increasing evidence suggests that THs also function in a variety of invertebrate species. Two alternative sources of hormone for animals are exogenous (from food items) and endogenous synthesis. We propose that exogenous THs can convey environmental information as well as regulate metabolism, revealing new communication avenues between organisms from different kingdoms. While such modes of cross-kingdom communication have been previously considered for fatty acid-based signaling and steroid hormones in plant-animal interactions, this is the first attempt to explore such a mode of action for TH signaling. We suggest that exogenous sources of TH (from food) may have been ancestral, while the ability to synthesize TH endogenously may have evolved independently in a variety of metazoans, resulting in a diversity of signaling pathways and, possibly, morphological structures involved in TH-signaling. PMID- 16339855 TI - The membrane-anchored BOTRYTIS-INDUCED KINASE1 plays distinct roles in Arabidopsis resistance to necrotrophic and biotrophic pathogens. AB - Plant resistance to disease is controlled by the combination of defense response pathways that are activated depending on the nature of the pathogen. We identified the Arabidopsis thaliana BOTRYTIS-INDUCED KINASE1 (BIK1) gene that is transcriptionally regulated by Botrytis cinerea infection. Inactivation of BIK1 causes severe susceptibility to necrotrophic fungal pathogens but enhances resistance to a virulent strain of the bacterial pathogen Pseudomonas syringae pv tomato. The response to an avirulent bacterial strain is unchanged, limiting the role of BIK1 to basal defense rather than race-specific resistance. The jasmonate and ethylene-regulated defense response, generally associated with resistance to necrotrophic fungi, is attenuated in the bik1 mutant based on the expression of the plant defensin PDF1.2 gene. bik1 mutants show altered root growth, producing more and longer root hairs, demonstrating that BIK1 is also required for normal plant growth and development. Whereas the pathogen responses of bik1 are mostly dependent on salicylic acid (SA) levels, the nondefense responses are independent of SA. BIK1 is membrane-localized, suggesting possible involvement in early stages of the recognition or transduction of pathogen response. Our data suggest that BIK1 modulates the signaling of cellular factors required for defense responses to pathogen infection and normal root hair growth, linking defense response regulation with that of growth and development. PMID- 16339857 TI - Transepithelial transport of fluorescent p-glycoprotein and MRP2 substrates by insect Malpighian tubules: confocal microscopic analysis of secreted fluid droplets. AB - Transport of fluorescent substrates of p-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (MRP2) by insect Malpighian tubules was examined using confocal laser scanning microscopy (CLSM). Isolated tubules of the cricket Teleogryllus commodus accumulated the MRP2 substrate Texas Red in the cells and lumen at concentrations up to 20 and 40 times, respectively, those in the bathing medium. Quantitative CLSM analysis of fluorochrome transport in some cricket tubules and all Drosophila tubules was not practical because of interfering effects of concretions in the cells and lumen. Samples of fluid secreted by tubules set up in Ramsay assays were therefore collected in hollow rectangle glass capillaries. Transepithelial dye flux was calculated as the product of fluid secretion rate (measured in the Ramsay assay) and dye concentration (measured by CLSM of the fluid samples). Dose-response curves for transport and the ratio of dye concentration in the secreted fluid to that in the bathing medium (S/M) were determined for Texas Red as well as for P-gp substrates (rhodamine 123, daunorubicin), the organic anion fluorescein and the organic cation quinacrine. Transepithelial transport of Texas Red was reduced by the MRP2 inhibitors MK571 and probenecid. Transport of daunorubicin was reduced by the P gp inhibitors verapamil and quinacrine and also by the organic cation tetraethylammonium. The results indicate the presence of P-gp-like and MRP2-like transporters in the Malpighian tubules of both species. PMID- 16339858 TI - Prediction of kinetics and kinematics of running animals using an analytical approximation to the planar spring-mass system. AB - The spring-mass model is often used to describe bouncing gaits. Although at first inspection the mechanical system appears simple, the solution to the motion cannot be derived easily. An analytical solution would provide a fast and intuitive method to determine the kinetic and kinematics of the centre of mass of terrestrial animals during over-ground steady state locomotion. Here, an analytical approximation using sine wave simplifications for the motion is presented. The analytical solution was almost indistinguishable from the numerical solution across initial leg angles of 17.5-30 degrees ; percentage differences between the analytical solution and the numerical solution were less than 1% for total mechanical energy, centre of mass position, total limb compression and centre of mass velocity and less than 2% different for resultant limb force and vertical acceleration of the centre of mass. The solution matched the relationship between stance time and speed collected from a trotting racehorse and accurately characterised previously published biological data. This study has shown that a simple analytical solution can predict the kinetics and kinematics of a spring-mass system over the range of biologically relevant sweep angles and horizontal velocities, and could be used to further understanding of limb deployment and gait selection. Using this analytical solution not only the force profile but also the changes in mechanical energy can be calculated from easily observed morphological and kinematic data. PMID- 16339859 TI - Chromatic interaction between egg pigmentation and skin chromatophores in the nuptial coloration of female two-spotted gobies. AB - In two-spotted gobies (Gobiusculus flavescens Fabricius 1779), females develop an orange belly as they approach sexual maturity. Bright belly coloration is preferred by males and has been suggested to act as a female ornament. This coloration is unusual in that it originates partly from pigmentation of the abdominal skin but also from strongly pigmented gonads directly visible through the skin. In addition, females have been observed to temporarily become more colourful during courtship and competition. To understand how gonad and skin pigmentation interact in this nuptial coloration, the potential for colour modification via regulation of skin chromatophores was investigated. Noradrenaline caused aggregation of chromatophore pigment and was used to experimentally reduce the contribution of skin chromatophores to the nuptial coloration. Chromatophore pigment aggregation caused bellies to become less colourful and abdominal skin biopsies to become less colourful and more transparent. There was a strong positive relationship between belly coloration and the coloration of the underlying gonads. This shows that belly coloration honestly reflects egg pigmentation, mainly because the transparency of the abdominal skin allows other fish to see the gonads directly. Interestingly, when noradrenaline caused pigment to aggregate and thereby increased the transparency of the skin, the relationship between belly and gonad coloration weakened. We conclude that female G. flavescens have a potential to use skin chromatophores to rapidly alter their nuptial coloration, thereby affecting the efficacy with which information about gonad coloration is conveyed. PMID- 16339860 TI - Echolocating bats can use acoustic landmarks for spatial orientation. AB - We investigated the echolocating bat's use of an acoustic landmark for orientation in a complex environment with no visual information. Three bats of the species Eptesicus fuscus were trained to fly through a hole in a mist net to receive a food reward on the other side. In all experiments, the vocal behavior of the bats was recorded simultaneously using a high-speed video recording system, allowing for a 3D reconstruction of the flight path. We ran three types of experiments, with different spatial relations between the landmark and net hole. In the first experiment, the bat's behavior was studied in test trials with the landmark placed 10 cm to the left of the net opening; between test trials, the positions of the net opening and landmark were moved, but the spatial relationship between the two remained fixed. With the landmark adjacent to the net opening, the bats quickly found the hole. In the second experiment, bats were tested in control trials in which the landmark was moved independently of the hole, breaking the established spatial relationship between the two. In control trials the bats repeatedly crashed into the net next to the landmark, and inspected the area around it. In the final experiment, the landmark was removed altogether from the set-up. Here the bats spent more time per trial searching for the net opening with an increased number of inspections as well as crashes into the net. However, over the course of a test day without the landmark, bats reduced the time spent per trial and focused inspections and crashes around the hole. The behavioral data show for the first time that the echolocating bat can learn to rely on an acoustic landmark to guide spatial orientation. PMID- 16339861 TI - Increase in intracellular pH induces phosphorylation of axonemal proteins for activation of flagellar motility in starfish sperm. AB - Increased intracellular pH ([pH]i) activates dynein in sea urchin and mammalian sperm and induces activation of flagellar motility. It is thought that cAMP dependent protein phosphorylation is associated with motility activation through increasing [pH]i, but little attention has been given to the cAMP-independent phosphorylation also induced by the [pH]i increase. The present study demonstrates that the increase in [pH]i in starfish sperm induces the phosphorylation of axonemal proteins and activation of flagellar motility independently of cAMP. Flagellar motility of intact sperm was activated when the [pH]i was raised by addition of NH4Cl. Histidine, which is known to activate motility of starfish sperm, also raised the [pH]i during the motility activation. In addition, motility of demembranated sperm flagella was activated in a pH dependent manner without cAMP. These results indicate that in starfish sperm it is the increase in [pH]i that induces activation of flagellar motility. Moreover, phosphorylation of axonemal proteins (of molecular mass 25, 32 and 45 kDa) was observed during the pH-dependent and cAMP-independent motility activation of demembranated sperm. This suggests that the increase in [pH]i regulates flagellar motility via cAMP-independent phosphorylation of axonemal proteins. PMID- 16339862 TI - An experimental study of social attraction and spacing between the sexes in sheep. AB - Most ungulates are gregarious species and outside the mating season are typically observed in single-sex groups. However little is known about the mechanisms underlying social segregation between sexes. We investigated the effect of conspecific attraction on individual spacing between unrestrained merino sheep Ovis aries and confined conspecifics. We considered differences between males and females and whether attractiveness of the confined conspecifics depends on their sex. A series of binary choice experiments was conducted in a large outdoor arena, located in pastures. One or two stimulus animals were placed in small individual cages (1.5 m x 1 m) on opposite sides of the arena. Sheep were tested with one fixed peer of the same or opposite sex vs an empty cage, and with two fixed peers of either the same sex as themselves, or one male and one female. Sheep in a control condition were exposed to two empty cages. In all of the test conditions, confined sheep were highly attractive. Males were more attracted by single stimulus peers of the same than the opposite sex, whereas females did not display such a preference. Sheep confronted with two restrained conspecifics tended to remain between the stimuli. This also occurred when the stimuli were of opposite sex, although the males tended to be located nearer the same-sex peer. Our findings can explain the strong aggregative behaviour of merino sheep, but also the social segregation previously observed in a mixed-sex group through higher attraction for same-sex than opposite-sex peers in males. PMID- 16339863 TI - Acute thermal stress and various heavy metals induce tissue-specific pro- or anti apoptotic events via the p38-MAPK signal transduction pathway in Mytilus galloprovincialis (Lam.). AB - We investigated the effects of various heavy metals such as copper, zinc and cadmium, as well as acute thermal stress, on the signalling mechanisms involved in the protection and/or apoptosis of Mytilus galloprovincialis mantle and gill tissues. The results of our studies revealed that mantle and gill tissues differentially respond to the stressful stimuli examined. In the mantle tissue, 1 micromol l(-1) Cu2+ and 50 micromol l(-1) Zn2+ induced a transient p38-MAPK activation, whereas 1 micromol l(-1) Cd2+ induced a biphasic profile of the kinase phosphorylation with maximal values at 15 and 120 min of treatment, respectively. Furthermore, 1 micromol l(-1) SB203580 abolished the Cu2+-induced kinase phosphorylation. In gills, both Cu2+ and Zn2+ induced a considerably higher p38-MAPK activation, which remained elevated for at least 60 min, whereas Cd2+ induced a maximal kinase activation within 60 min of treatment. Hypothermia (4 degrees C) induced a moderate kinase phosphorylation (maximised at 30 min), whereas hyperthermia (30 degrees C) induced a rapid (within 15 min) p38-MAPK phosphorylation that remained considerably above basal levels for at least 2 h. Our studies on the synergistic effect of hyperthermia and Cu2+ revealed that these two stressful stimuli are additive in the mantle tissue, inducing an almost double p38-MAPK activation. Further studies on the involvement of the p38-MAPK signalling pathway in tissue-specific pro- or anti-apoptotic events revealed that identical stressful stimuli possibly lead to apoptotic death via the caspase-3 activation in the mantle tissue and to anti-apoptotic events possibly via the induction of Hsp70 overexpression in the gill tissue. PMID- 16339864 TI - Plant senescence cues entry into diapause in the gall fly Eurosta solidaginis: resulting metabolic depression is critical for water conservation. AB - Mechanisms and possible cues for seasonal increases in desiccation resistance in larvae of the goldenrod gall fly Eurosta solidaginis, were examined before and after natural and premature plant senescence, or after being removed from their gall and placed in either 100, 95 or 75% relative humidity (RH). Rates of water loss were 8.6-fold lower, averaging 0.7+/-0.2 microg mm(-2) h(-1), in larvae from senescent gall tissue and after all RH treatments than in control larvae from pre senescent plants. Enhanced desiccation resistance occurred quickly, within 3 days of removal from their gall. Contrary to most previous reports, a large majority of the increased desiccation resistance (approximately 85%) was due to reduced respiratory transpiration with the remainder being the result of a lowered cuticular permeability. Rates of cuticular water loss were reduced by the presence of a vapor pressure gradient between the larval hemolymph and environmental water vapor and were probably due to increases in cuticular lipids and/or production of the cryoprotectant glycerol. Metabolic rate was reduced by over fourfold, averaging 0.07+/-0.01 microl CO2 g(-1) h(-1), in larvae from senescent gall tissue and all RH treatments compared to larvae from pre-senescent plants. The magnitude of the reduction in metabolic rates indicated that these larvae had entered diapause. In addition, larvae entered diapause in response to removal from, or degeneration of, the gall tissue they feed, on rather than seasonal changes in temperature or photoperiod. The low metabolic rates of the diapausing larvae probably allowed them to dramatically reduce their respiratory transpiration and total rate of water loss compared with non-diapausing controls. Thus, diapause, with its associated lowered metabolic rate, may be essential for conserving water in overwintering temperate insects, which may be dormant for six or more months of the year. PMID- 16339865 TI - The absence of mitochondrial DNA diversity among common laboratory inbred mouse strains. AB - Mitochondrial DNA (mtDNA), which exhibits a maternal inheritance and a high rate of evolution, has been widely used as a genetic marker when analyzing maternal lineage and inferring phylogenetic relationships among species. In this study, mtDNA variations among four classical (BALB/c, C3H, C57BL/6J and DBA/2) and three Chinese (TA2, 615 and T739) inbred strains of laboratory mice were analyzed by PCR-RFLP (polymerase chain reaction coupled with restriction fragment length polymorphism) and PCR-SSCP (polymerase chain reaction coupled with single stranded conformational polymorphism) techniques. PCR-RFLP analyses on 46 restriction sites revealed no variations in mtDNA D-loop (displacement loop), tRNA(Met+Glu+Ile) and ND3 (NADH dehydrogenase subunit 3) gene fragments in these strains. Furthermore, PCR-SSCP analyses demonstrated no variations in D-loop 5' and 3' end fragments in them. In view of enormous polymorphisms in mtDNA among mice and dramatic differences in nuclear genomes of these seven strains, our findings were surprising. However, in light of the maternal inheritance of mtDNA, the results indicate that the three Chinese strains, including TA2, T739 and 615, and the four classical strains, share a common maternal lineage. PMID- 16339866 TI - The insecticide pymetrozine selectively affects chordotonal mechanoreceptors. AB - Pymetrozine is a neuroactive insecticide but its site of action in the nervous system is unknown. Based on previous studies of symptoms in the locust, the feedback loop controlling the femur-tibia joint of the middle leg was chosen to examine possible targets of the insecticide. The femoral chordotonal organ, which monitors joint position and movement, turned out to be the primary site of pymetrozine action, while interneurons, motoneurons and central motor control circuitry in general did not noticeably respond to the insecticide. The chordotonal organs associated with the wing hinge stretch receptor and the tegula were influenced by pymetrozine in the same way as the femoral chordotonal organ, indicating that the insecticide affects chordotonal sensillae in general. Pymetrozine at concentrations down to 10(-8) mol l(-1) resulted in the loss of stimulus-related responses and either elicited (temporary) tonic discharges or eliminated spike activity altogether. Remarkably, pymetrozine affected the chordotonal organs in an all-or-none fashion, in agreement with previous independent studies. Other examined sense organs did not respond to pymetrozine, namely campaniform sensillae on the tibia and the subcosta vein, hair sensillae of the tegula (type I sensillae), and the wing hinge stretch receptor (type II sensillae). PMID- 16339867 TI - Comparative overwintering physiology of Alaska and Indiana populations of the beetle Cucujus clavipes (Fabricius): roles of antifreeze proteins, polyols, dehydration and diapause. AB - The beetle Cucujus clavipes is found in North America over a broad latitudinal range from North Carolina (latitude approximately 35 degrees N) to near tree line in the Brooks Range in Alaska (latitude, approximately 67 degrees 30' N). The cold adaptations of populations from northern Indiana (approximately 41 degrees 45' N) and Alaska were compared and, as expected, the supercooling points (the temperatures at which they froze) of these freeze-avoiding insects were significantly lower in Alaska insects. Both populations produce glycerol, but the concentrations in Alaska larvae were much higher than in Indiana insects (approximately 2.2 and 0.5 mol l(-1), respectively). In addition, both populations produce antifreeze proteins. Interestingly, in the autumn both populations have the same approximate level of hemolymph thermal hysteresis, indicative of antifreeze protein activity, suggesting that they synthesize similar amounts of antifreeze protein. A major difference is that the Alaska larvae undergo extreme dehydration in winter wherein water content decreases from 63-65% body water (1.70-1.85 g H2O g(-1) dry mass) in summer to 28-40% body water (0.40-0.68 g H2O g(-1) dry mass) in winter. These 2.5-4.6-fold reductions in body water greatly increase the concentrations of antifreeze in the Alaska insects. Glycerol concentrations would increase to 7-10 mol l(-1) while thermal hysteresis increased to nearly 13 degrees C (the highest ever measured in any organism) in concentrated hemolymph. By contrast, Indiana larvae do not desiccate in winter. The Alaska population also undergoes a diapause while insects from Indiana do not. The result of these, and likely additional, adaptations is that while the mean winter supercooling points of Indiana larvae were approximately -23 degrees C, those of Alaska larvae were -35 to -42 degrees C, and at certain times Alaska C. clavipes did not freeze when cooled to -80 degrees C. PMID- 16339868 TI - Locomotor function of the dorsal fin in rainbow trout: kinematic patterns and hydrodynamic forces. AB - In this study, we examine the kinematics and hydrodynamics of the soft dorsal fin in a representative basal teleost, the rainbow trout (Oncorhynchus mykiss), during steady rectilinear locomotion at 0.5-2.0 body lengths (L) s(-1) and during maneuvering. During steady swimming, dorsal fin height and sweep amplitude decrease with increasing speed. The dorsal fin wake, as viewed within a horizontal plane, consists of paired vortices on each side of the body (0.5 L s( 1)) or nearly linearly arrayed vortex centers above the body (1.0 L s(-1)) with central jet flows directed predominately laterally (lateral:thrust force ratio = 5-6). At 2.0 L s(-1), the dorsal fin is no longer recruited to add momentum to the wake. This pattern of decreasing involvement of the trout dorsal fin in thrust production with increasing speed contrasts with the results of our previous study of the soft dorsal fin of sunfish (Lepomis), which is hydrodynamically inactive at low speed and sheds a propulsive vortex wake at higher speed. Yawing maneuvers by trout involve unilateral production of a single vortex ring by the dorsal fin with a strong jet flow oriented almost directly laterally. During steady swimming, interception by the tail of the dorsal fin's vortical wake and the adipose fin's non-vortical (drag) wake is hypothesized as a mechanism for enhancing tail thrust. This study provides the first experimental evidence that the plesiomorphic soft dorsal fin of ray-finned fishes acts as an ancillary force generator during axial locomotion. We suggest that the distinction often made between median and paired fin (MPF) propulsion and body and caudal fin (BCF) propulsion in fishes obscures the important role of multiple propulsors acting in a coordinated fashion. Using a combination of anterior median fin oscillation and axial undulation, without continuous paired fin excursions, trout employ an ;M-BCF' gait during steady swimming. The primarily lateral orientation of dorsal fin force in trout induces corresponding roll and yaw moments, which must be countered by forces from the caudal, anal and paired fins. Locomotion in trout therefore involves the simultaneous active use of multiple fins, presumably to maintain body stability in the face of environmental perturbations. PMID- 16339869 TI - Insect gas exchange patterns: a phylogenetic perspective. AB - Most investigations of insect gas exchange patterns and the hypotheses proposed to account for their evolution have been based either on small-scale, manipulative experiments, or comparisons of a few closely related species. Despite their potential utility, no explicit, phylogeny-based, broad-scale comparative studies of the evolution of gas exchange in insects have been undertaken. This may be due partly to the preponderance of information for the endopterygotes, and its scarcity for the apterygotes and exopterygotes. Here we undertake such a broad-scale study. Information on gas exchange patterns for the large majority of insects examined to date (eight orders, 99 species) is compiled, and new information on 19 exemplar species from a further ten orders, not previously represented in the literature (Archaeognatha, Zygentoma, Ephemeroptera, Odonata, Mantodea, Mantophasmatodea, Phasmatodea, Dermaptera, Neuroptera, Trichoptera), is provided. These data are then used in a formal, phylogeny-based parsimony analysis of the evolution of gas exchange patterns at the order level. Cyclic gas exchange is likely to be the ancestral gas exchange pattern at rest (recognizing that active individuals typically show continuous gas exchange), and discontinuous gas exchange probably originated independently a minimum of five times in the Insecta. PMID- 16339870 TI - Ontogeny of bone strain: the zygomatic arch in pigs. AB - At the time of weaning, infant animals have little experience with hard food, and thus their skulls are not likely to be epigenetically adapted for the loads imposed by mastication. We examined bone strain in the zygomatic arch of 4-week old weanling piglets. Functional strains in piglets differed from those previously reported for older pigs in that the squamosal bone was not bent in the horizontal plane and the principal tensile strain on the zygomatic bone did not correspond to the direction of masseter muscle pull. Strain patterns were more variable in piglets than in older pigs. In older pigs, masticatory strains can be reproduced by stimulating the masseter muscles. When the piglet masseter was stimulated, strain patterns were more similar to those of older pigs, but shear strain magnitudes were the largest yet recorded from mammalian skull bones, up to 4000 muepsilon. To put these findings in the context of skeletal adaptation, 45 dry skulls, including some animals from the strain study, were measured. Reduced major axis regressions indicated that the infant arch was rounder in cross section and straighter than that of older animals. With growth, the arch became dorsoventrally higher, while mediolateral thickness decreased in the squamosal bone. Overall, these changes should make strain more predictable, explaining the lower variability in older animals. Other factors likely to be important in causing unique strain regimes in piglets include (1) unfamiliarity with hard food, (2) greater importance of muscles other than the same-side masseter and (3) greater proximity of molariform teeth to the arch. Collectively, these data indicate that the skeleton is not pre-adapted for specific functional loads. PMID- 16339871 TI - Flight behaviour during foraging of the social wasp Vespula vulgaris (Hymenoptera: Vespidae) and four mimetic hoverflies (Diptera: Syrphidae) Sericomyia silentis, Myathropa florea, Helophilus sp. and Syrphus sp. AB - Many hoverfly species show specific or non specific morphological resemblance to wasps (Vespula sp.) and it has been suggested that they also show similar flight behaviour. In this study we therefore compared the flight behaviour of wasps with that of four mimetic hoverflies, Sericomyia silentis, Myathropa florea, Helophilus sp. and Syrphus sp., by filming insects while they were foraging on an artificial array of flowers. Films were analysed to determine the routes taken, time spent hovering and flight speed. Of the four flies, only the non specific mimic, Syrphus, showed similar flight behaviour to the wasps; it flew more slowly, and with more roundabout routes than the other flies, hesitating before landing. These results suggest that in hoverflies there is little reason to expect strict correlation between morphological and behavioural mimicry; insects may acquire the similarities to their model more-or-less independently. PMID- 16339872 TI - Muscle fiber-type variation in lizards (Squamata) and phylogenetic reconstruction of hypothesized ancestral states. AB - Previously, we found that phrynosomatid lizards, a diverse group common in the southwestern USA, vary markedly in fiber-type composition of the iliofibularis (a hindlimb muscle important in locomotion). Phrynosomatidae comprises three subclades: the closely related sand and horned lizards, and their relatives the Sceloporus group. The variation in muscle fiber-type composition for 11 phrynosomatid species is attributable mainly to differences between the sand- and horned-lizard subclades. Here, we expand the phrynosomatid database with three additional species and compare these results with data collected for 10 outgroup (distantly related) species. Our goal was to determine if the patterns found in Phrynosomatidae hold across a broader phylogenetic range of the extant lizards and to elucidate the evolution of muscle fiber-type composition and related traits. To allow for meaningful comparisons, data were collected from species that are primarily terrestrial and relatively small in size (3.5-65 g body mass). Results indicate that the fiber-type variation observed within the Phrynosomatidae almost spans the range of variation observed in our sample of 24 species from eight families. However, one species of Acanthodactylus (Lacertidae) had a consistent region of large tonic fibers (that did not stain darkly for either succinic dehydrogenase or myosin ATPase activity), a fiber-type only occasionally seen in the other 23 species examined. Many species have a large proportion of either fast-twitch glycolytic (FG; e.g. sand lizards and Aspidoscelis) or fast-twitch oxidative-glycolytic (FOG) fibers (e.g. horned lizards), with the slow-oxidative proportion occupying only 1-17% of the iliofibularis. Importantly, the negative relationship between FG and FOG composition observed in Phrynosomatidae appears to be a characteristic of lizards in general, and could lead to functional trade-offs in aspects of locomotor performance, as has previously been reported for Lacertidae. Reconstruction of ancestral trait values by use of phylogenetically based statistical methods indicates especially large changes in fiber-type composition during the evolution of horned lizards. PMID- 16339873 TI - K+-independent initiation of motility in chum salmon sperm treated with an organic alcohol, glycerol. AB - Sperm of salmonid fishes are quiescent in the presence of millimolar concentrations of extracellular K+, but motility initiation occurs when sperm are suspended in K+-free medium. In this study, glycerol (CH2OHCHOHCH2OH) treatment of intact sperm in the presence of K+ induced the initiation of motility even though a large amount of K+ was present. Another organic alcohol, erythritol (CH2OH(CHOH)2CH2OH), had a similar effect, but ethylene glycol (CH2OHCH2OH) did not initiate sperm motility. Furthermore, this glycerol-treated sperm showed motility without subsequent addition of ATP and cAMP. CCCP, an uncoupler of the mitochondrial electron-transport chain involved in ATP synthesis, suppressed motility of glycerol-treated sperm, suggesting that ATP synthesis is required for dynein to slide microtubules in glycerol-treated sperm. The amount of intracellular cAMP ([cAMP]i) in glycerol-treated sperm did not increase on motility activation, but a protein kinase A (PKA) inhibitor, H-89, inhibited glycerol-treated sperm motility. In addition, phosphorylation of protein associated with motility initiation also occurred in glycerol-treated sperm, suggesting that the glycerol treatment induces activation of PKA without an increase in [cAMP]i. Taken together, it can be concluded that organic alcohol, glycerol and erythritol induce phosphorylation for motility initiation, bypassing the increase in [cAMP]i as a result of a decrease in extracellular K+ concentration. PMID- 16339874 TI - An X-Ray diffraction study on mouse cardiac cross-bridge function in vivo: effects of adrenergic {beta}-stimulation. AB - To investigate how beta-stimulation affects the contractility of cardiac muscle, x-ray diffraction from cardiac muscle in the left ventricular free wall of a mouse heart was recorded in vivo. To our knowledge, this is the first x-ray diffraction study on a heart in a living body. After the R wave in electrocardiograms, the ratio of the intensities of the equatorial (1,0) and (1,1) reflections decreased for approximately 50 ms from a diastolic value of 2.1 to a minimum of 0.8, and then recovered. The spacing of the (1,0) lattice planes increased for approximately 90 ms from a diastolic value of 37.2 nm to a maximum of 39.1 nm, and then returned to the diastolic level, corresponding to approximately 10% stretch of sarcomere. Stimulation of beta-adrenergic receptor by dobutamine (20 microg/kg/min) accelerated both the decrease in the intensity ratio, which reached a smaller systolic value, and the increase in the lattice spacing. However, the intensity ratio and spacing at the end-diastole were unchanged. The recovery of the lattice spacing during relaxation was also accelerated. The mass transfer to the thin filaments at systole in a beta stimulated heart was close to the peak value in twitch of frog skeletal muscle at 4 degrees C, showing that the majority of cross-bridges have been recruited with few in reserve. PMID- 16339875 TI - Rigidity sensing at the leading edge through alphavbeta3 integrins and RPTPalpha. AB - Cells require optimal substrate stiffness for normal function and differentiation. The mechanisms for sensing matrix rigidity and durotaxis, however, are not clear. Here we showed that control, Shp2-/-, integrin beta1-/-, and talin1-/- cell lines all spread to a threefold greater area on fibronectin (FN)-coated rigid polyacrylamide surfaces than soft. In contrast, RPTPalpha-/- cells spread to the same area irrespective of rigidity on FN surfaces but spread 3x greater on rigid collagen IV-coated surfaces than soft. RPTPalpha and alphavbeta3 integrins were shown previously to be colocalized at leading edges and antibodies to alphavbeta3 blocked FN rigidity sensing. When FN beads were held with a rigid laser trap at the leading edge, stronger bonds to the cytoskeleton formed than when held with a soft trap; whereas back from the leading edge and in RPTPalpha-/- cells, weaker bonds were formed with both rigid and soft laser traps. From the rigidity of the trap, we calculate that a force of 10 pN generated in 1 s is sufficient to activate the rigidity response. We suggest that RPTPalpha and alphavbeta3 at the leading edge are critical elements for sensing FN matrix rigidity possibly through SFK activation at the edge and downstream signaling. PMID- 16339876 TI - Structure of core domain of fibril-forming PHF/Tau fragments. AB - Short peptide sequences within the microtubule binding domain of the protein Tau are proposed to be core nucleation sites for formation of amyloid fibrils displaying the paired helical filament (PHF) morphology characteristic of neurofibrillary tangles. To study the structure of these proposed nucleation sites, we analyzed the x-ray diffraction patterns from the assemblies formed by a variety of PHF/tau-related peptide constructs containing the motifs VQIINK (PHF6*) in the second repeat and VQIVYK (PHF6) in the third repeat of tau. Peptides included: tripeptide acetyl-VYK-amide (AcVYK), tetrapeptide acetyl-IVYK amide (AcPHF4), hexapeptide acetyl-VQIVYK-amide (AcPHF6), and acetyl GKVQIINKLDLSNVQKDNIKHGSVQIVYKPVDLSKVT-amide (AcTR4). All diffraction patterns showed reflections at spacings of 4.7 A, 3.8 A, and approximately 8-10 A, which are characteristic of an orthogonal unit cell of beta-sheets having dimensions a=9.4 A, b=6.6 A, and c=approximately 8-10 A (where a, b, and c are the lattice constants in the H-bonding, chain, and intersheet directions). The sharp 4.7 A reflections indicate that the beta-crystallites are likely to be elongated along the H-bonding direction and in a cross-beta conformation. The assembly of the AcTR4 peptide, which contains both the PHF6 and PHF6* motifs, consisted of twisted sheets, as indicated by a unique fanning of the diffuse equatorial scattering and meridional accentuation of the (210) reflection at 3.8 A spacing. The diffraction data for AcVYK, AcPHF4, and AcPHF6 all were consistent with approximately 50 A-wide tubular assemblies having double-walls, where beta strands constitute the walls. In this structure, the peptides are H-bonded together in the fiber direction, and the intersheet direction is radial. The positive-charged lysine residues face the aqueous medium, and tyrosine-tyrosine aromatic interactions stabilize the intersheet (double-wall) layers. This particular contact, which may be involved in PHF fibril formation, is proposed here as a possible aromatic target for anti-tauopathy drugs. PMID- 16339877 TI - Evaluating tilt angles of membrane-associated helices: comparison of computational and NMR techniques. AB - A computational method to calculate the orientation of membrane-associated alpha helices with respect to a lipid bilayer has been developed. It is based on a previously derived implicit membrane representation, which was parameterized using the structures of 46 alpha-helical membrane proteins. The method is validated by comparison with an independent data set of six transmembrane and nine antimicrobial peptides of known structure and orientation. The minimum energy orientations of the transmembrane helices were found to be in good agreement with tilt and rotation angles known from solid-state NMR experiments. Analysis of the free-energy landscape found two types of minima for transmembrane peptides: i), Surface-bound configurations with the helix long axis parallel to the membrane, and ii), inserted configurations with the helix spanning the membrane in a perpendicular orientation. In all cases the inserted configuration also contained the global energy minimum. Repeating the calculations with a set of solution NMR structures showed that the membrane model correctly distinguishes native transmembrane from nonnative conformers. All antimicrobial peptides investigated were found to orient parallel and bind to the membrane surface, in agreement with experimental data. In all cases insertion into the membrane entailed a significant free-energy penalty. An analysis of the contributions of the individual residue types confirmed that hydrophobic residues are the main driving force behind membrane protein insertion, whereas polar, charged, and aromatic residues were found to be important for the correct orientation of the helix inside the membrane. PMID- 16339878 TI - Modeling the VPAC2-activated cAMP/PKA signaling pathway: from receptor to circadian clock gene induction. AB - Increasing evidence suggests an important role for VPAC2-activated signal transduction pathways in maintaining a synchronized biological clock in the suprachiasmatic nucleus (SCN). Activation of the VPAC2 signaling pathway induces per1 gene expression in the SCN and phase-shifts the circadian clock. Mice without the VPAC2 receptor lack an overt, coherent circadian rhythm in clock gene expression, SCN neuron firing rate, and locomotor behavior. Using a systems approach, we have developed a kinetic model integrating VPAC2 signaling mediated by the cyclic AMP (cAMP)/protein kinase A (PKA) pathway and leading to induced circadian clock gene expression. We fit the model to experimental data from the literature for cAMP accumulation, PKA activation, cAMP-response element binding protein phosphorylation, and per1 induction. By linking the VPAC2 model to a published circadian clock model, we also simulated clock phase shifts induced by vasoactive intestinal polypeptide (VIP) and matched experimental data for the VIP response. The simulated phase response curve resembled the hamster response to a related neuropeptide, GRP1-27, and light. Simulations using pulses of VIP revealed that the system response is extraordinarily robust to input signal duration, a result with physiologically relevant consequences. Lastly, simulations using varied receptor levels matched literature experimental data from animals overexpressing VPAC2 receptors. PMID- 16339879 TI - Flexural rigidity of individual microtubules measured by a buckling force with optical traps. AB - We used direct buckling force measurements with optical traps to determine the flexural rigidity of individual microtubules bound to polystyrene beads. To optimize the accuracy of the measurement, we used two optical traps and antibody coated beads to manipulate each microtubule. We then applied a new analytical model assuming nonaxial buckling. Paclitaxel-stabilized microtubules were polymerized from purified tubulin, and the average microtubule rigidity was calculated as 2.0 x 10(-24) Nm2 using this novel microtubule buckling system. This value was not dependent on microtubule length. We also measured the rigidity of paclitaxel-free microtubules, and obtained the value of 7.9 x 10(-24) Nm2, which is nearly four times that measured for paclitaxel-stabilized microtubules. PMID- 16339880 TI - Neuronal SNAREs do not trigger fusion between synthetic membranes but do promote PEG-mediated membrane fusion. AB - At low surface concentrations that permit formation of impermeable membranes, neuronal soluble N-ethyl maleimide sensitive factor attachment protein receptor (SNARE) proteins form a stable, parallel, trans complex when vesicles are brought into contact by a low concentration of poly(ethylene glycol) (PEG). Surprisingly, formation of a stable SNARE complex does not trigger fusion under these conditions. However, neuronal SNAREs do promote fusion at low protein/lipid ratios when triggered by higher concentrations of PEG. Promotion of PEG-triggered fusion required phosphatidylserine and depended only on the surface concentration of SNAREs and not on the formation of a trans SNARE complex. These results were obtained at protein surface concentrations reported for synaptobrevin in synaptic vesicles and with an optimally fusogenic lipid composition. At a much higher protein/lipid ratio, vesicles joined by SNARE complex slowly mixed lipids at 37 degrees C in the absence of PEG, in agreement with earlier reports. However, vesicles containing syntaxin at a high protein/lipid ratio (>or=1:250) lost membrane integrity. We conclude that the neuronal SNARE complex promotes fusion by joining membranes and that the individual proteins syntaxin and synaptobrevin disrupt membranes so as to favor formation of a stalk complex and to promote conversion of the stalk to a fusion pore. These effects are similar to the effects of viral fusion peptides and transmembrane domains, but they are not sufficient by themselves to produce fusion in our in vitro system at surface concentrations documented to occur in synaptic vesicles. Thus, it is likely that proteins or factors other than the SNARE complex must trigger fusion in vivo. PMID- 16339881 TI - A quantitative analysis of cardiac myocyte relaxation: a simulation study. AB - The determinants of relaxation in cardiac muscle are poorly understood, yet compromised relaxation accompanies various pathologies and impaired pump function. In this study, we develop a model of active contraction to elucidate the relative importance of the [Ca2+]i transient magnitude, the unbinding of Ca2+ from troponin C (TnC), and the length-dependence of tension and Ca2+ sensitivity on relaxation. Using the framework proposed by one of our researchers, we extensively reviewed experimental literature, to quantitatively characterize the binding of Ca2+ to TnC, the kinetics of tropomyosin, the availability of binding sites, and the kinetics of crossbridge binding after perturbations in sarcomere length. Model parameters were determined from multiple experimental results and modalities (skinned and intact preparations) and model results were validated against data from length step, caged Ca2+, isometric twitches, and the half-time to relaxation with increasing sarcomere length experiments. A factorial analysis found that the [Ca2+]i transient and the unbinding of Ca2+ from TnC were the primary determinants of relaxation, with a fivefold greater effect than that of length-dependent maximum tension and twice the effect of tension-dependent binding of Ca2+ to TnC and length-dependent Ca2+ sensitivity. The affects of the [Ca2+]i transient and the unbinding rate of Ca2+ from TnC were tightly coupled with the effect of increasing either factor, depending on the reference [Ca2+]i transient and unbinding rate. PMID- 16339883 TI - The kinetics of analyte capture on nanoscale sensors. AB - This article presents a number of kinetic analyses related to binding processes relevant to capture of target analyte species in nanoscale cantilever-type devices designed to detect small concentrations of biomolecules. The overall analyte capture efficiency is a crucial measure of the ultimate sensitivity of such devices, and a detailed kinetic analysis tells us how rapidly such measurements may be made. We have analyzed the capture kinetics under a variety of conditions, including the possibility of so-called surface-enhanced ligand capture. One of the modalities studied requires ligand capture through a cross linking mechanism, and it was found that this mode may provide a robust and sensitive approach to biomolecular detection. For the two modalities studied, we find that detection of specific biomolecules down to concentration levels of 1 nM or less appear to be quite feasible for the device configurations studied. PMID- 16339882 TI - Bistability in apoptosis: roles of bax, bcl-2, and mitochondrial permeability transition pores. AB - We propose a mathematical model for mitochondria-dependent apoptosis, in which kinetic cooperativity in formation of the apoptosome is a key element ensuring bistability. We examine the role of Bax and Bcl-2 synthesis and degradation rates, as well as the number of mitochondrial permeability transition pores (MPTPs), on the cell response to apoptotic stimuli. Our analysis suggests that cooperative apoptosome formation is a mechanism for inducing bistability, much more robust than that induced by other mechanisms, such as inhibition of caspase 3 by the inhibitor of apoptosis (IAP). Simulations predict a pathological state in which cells will exhibit a monostable cell survival if Bax degradation rate is above a threshold value, or if Bax expression rate is below a threshold value. Otherwise, cell death or survival occur depending on initial caspase-3 levels. We show that high expression rates of Bcl-2 can counteract the effects of Bax. Our simulations also demonstrate a monostable (pathological) apoptotic response if the number of MPTPs exceeds a threshold value. This study supports our contention, based on mathematical modeling, that cooperativity in apoptosome formation is critically important for determining the healthy responses to apoptotic stimuli, and helps define the roles of Bax, Bcl-2, and MPTP vis-a-vis apoptosome formation. PMID- 16339884 TI - Temperature dependence of the volumetric parameters of drug binding to poly[d(A T)].Poly[d(A-T)] and Poly(dA).Poly(dT). AB - We report the temperature and salt dependence of the volume change (DeltaVb) associated with the binding of ethidium bromide and netropsin with poly(dA).poly(dT) and poly[d(A-T)].poly[d(A-T)]. The DeltaV(b) of binding of ethidium with poly(dA).poly(dT) was much more negative at temperatures approximately 70 degrees C than at 25 degrees C, whereas the difference is much smaller in the case of binding with poly[d(A-T)].poly[d(A-T)]. We also determined the volume change of DNA-drug interaction by comparing the volume change of melting of DNA duplex and DNA-drug complex. The DNA-drug complexes display helix coil transition temperatures (Tm several degrees above those of the unbound polymers, e.g., the Tm of the netropsin complex with poly(dA)poly(dT) is 106 degrees C. The results for the binding of ethidium with poly[d(A-T)].poly[d(A-T)] were accurately described by scaled particle theory. However, this analysis did not yield results consistent with our data for ethidium binding with poly(dA).poly(dT). We hypothesize that heat-induced changes in conformation and hydration of this polymer are responsible for this behavior. The volumetric properties of poly(dA).poly(dT) become similar to those of poly[d(A-T)].poly[d(A T)] at higher temperatures. PMID- 16339885 TI - Redox regulation of CLIC1 by cysteine residues associated with the putative channel pore. AB - Chloride intracellular channels (CLICs) are putative pore-forming glutathione-S transferase homologs that are thought to insert into cell membranes directly from the cytosol. We incorporated soluble, recombinant human CLIC1 into planar lipid bilayers to investigate the associated ion channels, and noted that channel assembly (unlike membrane insertion) required a specific lipid mixture. The channels formed by reduced CLIC1 were similar to those previously recorded from cells and "tip-dip" bilayers, and specific anti-CLIC1 antibodies inhibited them. However, the amplitudes of the filtered single-channel currents were strictly regulated by the redox potential on the "extracellular" (or "luminal") side of the membrane, with minimal currents under strongly oxidizing conditions. We carried out covalent functional modification and site-directed mutagenesis of this controversial ion channel to test the idea that cysteine 24 is a critical redox-sensitive residue located on the extracellular (or luminal) side of membrane CLIC1 subunits, in a cysteine-proline motif close to the putative channel pore. Our findings support a simple structural hypothesis to explain how CLIC1 oligomers form pores in membranes, and suggest that native channels may be regulated by a novel mechanism involving the formation and reduction of intersubunit disulphide bonds. PMID- 16339886 TI - Temperature dependence and thermodynamics of Klenow polymerase binding to primed template DNA. AB - DNA binding of Klenow polymerase has been characterized with respect to temperature to delineate the thermodynamic driving forces involved in the interaction of this polymerase with primed-template DNA. The temperature dependence of the binding affinity exhibits distinct curvature, with tightest binding at 25-30 degrees C. Nonlinear temperature dependence indicates Klenow binds different primed-template constructs with large heat capacity (DeltaCp) values (-870 to -1220 cal/mole K) and thus exhibits large temperature dependent changes in enthalpy and entropy. Binding is entropy driven at lower temperatures and enthalpy driven at physiological temperatures. Large negative DeltaCp values have been proposed to be a 'signature' of site-specific DNA binding, but type I DNA polymerases do not exhibit significant DNA sequence specificity. We suggest that the binding of Klenow to a specific DNA structure, the primed-template junction, results in a correlated thermodynamic profile that mirrors what is commonly seen for DNA sequence-specific binding proteins. Klenow joins a small number of other DNA-sequence independent DNA binding proteins which exhibit unexpectedly large negative DeltaCp values. Spectroscopic measurements show small conformational rearrangements of both the DNA and Klenow upon binding, and small angle x-ray scattering shows a global induced fit conformational compaction of the protein upon binding. Calculations from both crystal structure and solution structural data indicate that Klenow DNA binding is an exception to the often observed correlation between DeltaCp and changes in accessible surface area. In the case of Klenow, surface area burial can account for only about half of the DeltaCp of binding. PMID- 16339887 TI - Structural characterization and pH-induced conformational transition of full length KcsA. AB - The bacterial K+ channel KcsA from Streptomyces lividans was analyzed by neutron and x-ray small-angle solution scattering. The C-terminally truncated version of KcsA, amenable to crystallographic studies, was compared with the full-length channel. Analyzing the scattering data in terms of radius of gyration reveals differences between both KcsA species of up to 13.2 A. Equally, the real-space distance distribution identifies a 40 to 50 A extension of full-length KcsA compared to its C-terminally truncated counterpart. We show that the x-ray and neutron scattering data are amenable for molecular shape reconstruction of full length KcsA. The molecular envelopes calculated display an hourglass-shaped structure within the C-terminal intracellular domain. The C-terminus extends the membrane spanning region of KcsA by 54-70 A, with a central constriction 10-30 A wide. Solution scattering techniques were further employed to characterize the KcsA channel under acidic conditions favoring its open conformation. The full length KcsA at pH 5.0 shows the characteristics of a dumbbell-shaped macromolecular structure, originating from dimerization of the tetrameric K+ channel. Since C-terminally truncated KcsA measured under the same low pH conditions remains tetrameric, oligomerization of full-length KcsA seems to proceed via structurally changed C-terminal domains. The determined maximum dimensions of the newly formed complex increase by 50-60%. Shape reconstruction of the pseudooctameric complex indicates the pH-induced conformational reorganization of the intracellular C-terminal domain. PMID- 16339888 TI - Measuring limits of telomere movement on nuclear envelope. AB - The dynamic behavior of the decondensed chromatin can be monitored by real-time fluorescence confocal microscopy. It can be observed that different chromosomal sites enjoy different degrees of freedom during a certain period, exploring larger or smaller portions of nuclear volume. Here we measure the accessible surface for two chromosomal sites (yeast telomeres Tel3R and Tel6R) that both exhibit strong preferential association with the nuclear membrane in galactose containing media, but differ significantly in gene activity. Telomere Tel6R, which harbors an inducible gene with high levels of transcription, explores a much larger surface than the telomere Tel3R, which is adjacent to inactive chromatin. Thus, our results distinguish two perinuclear movements characteristic of different transcriptional state, allowing for a better understanding of the correlation between activity of genes and chromatin dynamics. PMID- 16339889 TI - Interaction of zwitterionic penicillins with the OmpF channel facilitates their translocation. AB - To study translocation of beta-lactam antibiotics of different size and charge across the outer bacterial membrane, we combine an analysis of ion currents through single trimeric outer membrane protein F (OmpF) porins in planar lipid bilayers with molecular dynamics simulations. Because the size of penicillin molecules is close to the size of the narrowest part of the OmpF pore, penicillins occlude the pore during their translocation. Favorably interacting penicillins cause time-resolvable transient blockages of the small-ion current through the channel and thereby provide information about their dynamics within the pore. Analyzing these random fluctuations, we find that ampicillin and amoxicillin have a relatively high affinity for OmpF. In contrast, no or only a weak interaction is detected for carbenicillin, azlocillin, and piperacillin. Molecular dynamics simulations suggest a possible pathway of these drugs through the OmpF channel and rationalize our experimental findings. For zwitterionic ampicillin and amoxicillin, we identify a region of binding sites near the narrowest part of the channel pore. Interactions with these sites partially compensate for the entropic cost of drug confinement by the channel. Whereas azlocillin and piperacillin are clearly too big to pass through the channel constriction, dianionic carbenicillin does not find an efficient binding region in the constriction zone. Carbenicillin's favorable interactions are limited to the extracellular vestibule. These observations confirm our earlier suggestion that a set of high-affinity sites at the narrowest part of the OmpF channel improves a drug's ability to cross the membrane via the pore. PMID- 16339890 TI - Solid-state NMR analysis of the PGLa peptide orientation in DMPC bilayers: structural fidelity of 2H-labels versus high sensitivity of 19F-NMR. AB - The structure and alignment of the amphipathic alpha-helical antimicrobial peptide PGLa in a lipid membrane is determined with high accuracy by solid-state 2H-NMR. Orientational constraints are derived from a series of eight alanine 3,3,3-d3-labeled peptides, in which either a native alanine is nonperturbingly labeled (4x), or a glycine (2x) or isoleucine (2x) is selectively replaced. The concentration dependent realignment of the alpha-helix from the surface-bound "S state" to a tilted "T-state" by 30 degrees is precisely calculated using the quadrupole splittings of the four nonperturbing labels as constraints. The remaining, potentially perturbing alanine-3,3,3-d3 labels show only minor deviations from the unperturbed peptide structure and help to single out the unique solution. Comparison with previous 19F-NMR constraints from 4-CF3 phenylglycine labels shows that the structure and orientation of the PGLa peptide is not much disturbed even by these bulky nonnatural side chains, which contain CF3 groups that offer a 20-fold better NMR sensitivity than CD3 groups. PMID- 16339891 TI - A FRET-based calcium biosensor with fast signal kinetics and high fluorescence change. AB - Genetically encoded calcium biosensors have become valuable tools in cell biology and neuroscience, but some aspects such as signal strength and response kinetics still need improvement. Here we report the generation of a FRET-based calcium biosensor employing troponin C as calcium-binding moiety that is fast, is stable in imaging experiments, and shows a significantly enhanced fluorescence change. These improvements were achieved by engineering magnesium and calcium-binding properties within the C-terminal lobe of troponin C and by the incorporation of circularly permuted variants of the green fluorescent protein. This sensor named TN-XL shows a maximum fractional fluorescence change of 400% in its emission ratio and linear response properties over an expanded calcium regime. When imaged in vivo at presynaptic motoneuron terminals of transgenic fruit flies, TN-XL exhibits highly reproducible fluorescence signals with the fastest rise and decay times of all calcium biosensors known so far. PMID- 16339892 TI - Activation of naive B lymphocytes via CD81, a pathogenetic mechanism for hepatitis C virus-associated B lymphocyte disorders. AB - Infection with hepatitis C virus (HCV), a leading cause of chronic liver diseases, can associate with B lymphocyte proliferative disorders, such as mixed cryoglobulinemia and non-Hodgkin lymphoma. The major envelope protein of HCV (HCV E2) binds, with high affinity CD81, a tetraspanin expressed on several cell types. Here, we show that engagement of CD81 on human B cells by a combination of HCV-E2 and an anti-CD81 mAb triggers the JNK pathway and leads to the preferential proliferation of the naive (CD27-) B cell subset. In parallel, we have found that B lymphocytes from the great majority of chronic hepatitis C patients are activated and that naive cells display a higher level of activation markers than memory (CD27+) B lymphocytes. Moreover, eradication of HCV infection by IFN therapy is associated with normalization of the activation-markers expression. We propose that CD81-mediated activation of B cells in vitro recapitulates the effects of HCV binding to B cell CD81 in vivo and that polyclonal proliferation of naive B lymphocytes is a key initiating factor for the development of the HCV-associated B lymphocyte disorders. PMID- 16339893 TI - Solution structure of a multifunctional DNA- and protein-binding motif of human Werner syndrome protein. AB - Werner syndrome (WS) is an autosomal recessive disease that results in premature aging. Mutations in the WS gene (WRN) result in a loss of expression of the WRN protein and predispose WS patients to accelerated aging. As a helicase and a nuclease, WRN is unique among the five human RecQ helicase family members and is capable of multiple functions involved in DNA replication, repair, recombination, and telomere maintenance. A 144-residue fragment of WRN was previously determined to be a multifunctional DNA- and protein-binding domain (DPBD) that interacts with structure-specific DNA and a variety of DNA-processing proteins. In addition, DPBD functions as a nucleolar targeting sequence of WRN. The solution structure of the DPBD, the first of a WRN fragment, has been solved by NMR. DPBD consists of a winged helix-like motif and an unstructured C-terminal region of approximately 20 aa. The putative DNA-binding surface of DPBD has been identified by using known structural and biochemical data. Based on the structural data and on the biochemical data, we suggest a surface on the DPBD for interacting with other proteins. In this structural model, a single winged helix domain binds to both DNA and other proteins. Furthermore, we propose that DPBD functions as a regulatory domain to regulate the enzymatic activity of WRN and to direct cellular localization of WRN through protein-protein interaction. PMID- 16339894 TI - Neurons of the cerebral cortex exhibit precise interspike timing in correspondence to behavior. AB - We show that times of spikes can be very precise. In the cerebral cortex, where each nerve cell is affected by thousands of others, it is the common belief that the exact time of a spike is random up to an averaged firing rate over tens of milliseconds. In a brain slice, precise time relations of several neurons have been observed. It remained unclear whether this phenomenon can also be observed in brains of behaving animals. Here we show, in behaving monkeys, that time intervals between spikes, measured in correspondence to a specific behavior, may be controlled to within the milliseconds range. PMID- 16339895 TI - A high-resolution whole-genome cattle-human comparative map reveals details of mammalian chromosome evolution. AB - Approximately 3,000 cattle bacterial artificial chromosome (BAC)-end sequences were added to the Illinois-Texas 5,000-rad RH (RH, radiation hybrid) map. The BAC end sequences selected for mapping are approximately 1 Mbp apart on the human chromosomes as determined by blastn analysis. The map has 3,484 ordered markers, of which 3,204 are anchored in the human genome. Two hundred-and-one homologous synteny blocks (HSBs) were identified, of which 27 are previously undiscovered, 79 are extended, 26 were formed by previously unrecognized breakpoints in 18 previously defined HSBs, and 23 are the result of fusions. The comparative coverage relative to the human genome is approximately 91%, or 97% of the theoretical maximum. The positions of 64% of all cattle centromeres and telomeres were reassigned relative to their positions on the previous map, thus facilitating a more detailed comparative analysis of centromere and telomere evolution. As an example of the utility of the high-resolution map, 22 cattle BAC fingerprint contigs were directly anchored to cattle chromosome 19 [Bos taurus, (BTA) 19]. The order of markers on the cattle RH and fingerprint maps of BTA19 and the sequence-based map of human chromosome 17 [Homo sapiens, (HSA) 17] were found to be highly consistent, with only two minor ordering discrepancies between the RH map and fingerprint contigs. The high-resolution Illinois-Texas 5,000-rad RH and comparative maps will facilitate identification of candidate genes for economically important traits, the phylogenomic analysis of mammalian chromosomes, proofing of the BAC fingerprint map and, ultimately, aid the assembly of cattle whole-genome sequence. PMID- 16339896 TI - Solvent and mutation effects on the nucleation of amyloid beta-protein folding. AB - Experimental evidence suggests that the folding and aggregation of the amyloid beta-protein (Abeta) into oligomers is a key pathogenetic event in Alzheimer's disease. Inhibiting the pathologic folding and oligomerization of Abeta could be effective in the prevention and treatment of Alzheimer's disease. Here, using all atom molecular dynamics simulations in explicit solvent, we probe the initial stages of folding of a decapeptide segment of Abeta, Abeta(21-30), shown experimentally to nucleate the folding process. In addition, we examine the folding of a homologous decapeptide containing an amino acid substitution linked to hereditary cerebral hemorrhage with amyloidosis-Dutch type, [Gln-22]Abeta(21 30). We find that: (i) when the decapeptide is in water, hydrophobic interactions and transient salt bridges between Lys-28 and either Glu-22 or Asp-23 are important in the formation of a loop in the Val-24-Lys-28 region of the wild-type decapeptide; (ii) in the presence of salt ions, salt bridges play a more prominent role in the stabilization of the loop; (iii) in water with a reduced density, the decapeptide forms a helix, indicating the sensitivity of folding to different aqueous environments; and (iv) the "Dutch" peptide in water, in contrast to the wild-type peptide, fails to form a long-lived Val-24-Lys-28 loop, suggesting that loop stability is a critical factor in determining whether Abeta folds into pathologic structures. PMID- 16339897 TI - Autoreactive CD8 T cells associated with beta cell destruction in type 1 diabetes. AB - Type 1 diabetes is a T cell-mediated autoimmune disease, and insulin is an important target of the autoimmune response associated with beta cell destruction. The mechanism of destruction is still unknown. Here, we provide evidence for CD8 T cell autoreactivity associated with recurrent autoimmunity and loss of beta cell function in type 1 diabetic islet transplant recipients. We first identified an insulin B chain peptide (insB10-18) with extraordinary binding affinity to HLA-A2(*0201) that is expressed by the majority of type 1 diabetes patients. We next demonstrated that this peptide is naturally processed by both constitutive and immuno proteasomes and translocated to the endoplasmic reticulum by the peptide transporter TAP1 to allow binding to HLA-A2 in the endoplasmic reticulum and cell surface presentation. Peripheral blood mononuclear cells from a healthy donor were primed in vitro with this peptide, and CD8 T cells were isolated that specifically recognize target cells expressing the insulin B chain peptide. HLA-A2(insB10-18) tetramer staining revealed a strong association between detection of autoreactive CD8 T cells and recurrent autoimmunity after islet transplantation and graft failure in type 1 diabetic patients. We demonstrate that CD8 T cell autoreactivity is associated with beta cell destruction in type 1 diabetes in humans. PMID- 16339898 TI - A selective metabotropic glutamate receptor 7 agonist: activation of receptor signaling via an allosteric site modulates stress parameters in vivo. AB - Metabotropic glutamate receptor (mGluR) subtypes (mGluR1 to mGluR8) act as important pre- and postsynaptic regulators of neurotransmission in the CNS. These receptors consist of two domains, an extracellular region containing the orthosteric agonist site and a transmembrane heptahelical domain involved in G protein activation and recognition of several recently synthesized pharmacological modulators. The presynaptic receptor mGluR7 shows the highest evolutionary conservation within the family, but no selective pharmacological tool was known. Here we characterize an mGluR7-selective agonist, N,N' dibenzhydrylethane-1,2-diamine dihydrochloride (AMN082), which directly activates receptor signaling via an allosteric site in the transmembrane domain. At transfected mammalian cells expressing mGluR7, AMN082 potently inhibits cAMP accumulation and stimulates GTPgammaS binding (EC50-values, 64-290 nM) with agonist efficacies comparable with those of L-2-amino-4-phosphonobutyrate (L-AP4) and superior to those of L-glutamate. AMN082 (< or = 10 microM) failed to show appreciable activating or inhibitory effects at other mGluR subtypes and selected ionotropic GluRs. Chimeric receptor studies position the binding site of AMN082 in the transmembrane region of mGluR7, and we demonstrate that this allosteric agonist has little, if any, effect on the potency of orthosteric ligands. Here we provide evidence for full agonist activity mediated by the heptahelical domain of family 3 G protein-coupled receptors (which have mGluR-like structure) that may lead to drug development opportunities. Further, AMN082 is orally active, penetrates the blood-brain barrier, and elevates the plasma stress hormones corticosterone and corticotropin in an mGluR7-dependent fashion. Therefore, AMN082 is a valuable tool for unraveling the role of mGluR7 in stress-related CNS disorders. PMID- 16339899 TI - Bordetella filamentous hemagglutinin plays a critical role in immunomodulation, suggesting a mechanism for host specificity. AB - Bordetella pertussis, the causative agent of the acute childhood respiratory disease whooping cough, is a human-adapted variant of Bordetella bronchiseptica, which displays a broad host range and typically causes chronic, asymptomatic infections. These pathogens express a similar but not identical surface-exposed and secreted protein called filamentous hemagglutinin (FHA) that has been proposed to function as both a primary adhesin and an immunomodulator. To test the hypothesis that FHA plays an important role in determining host specificity and/or the propensity to cause acute versus chronic disease, we constructed a B. bronchiseptica strain expressing FHA from B. pertussis (FHA(Bp)) and compared it with wild-type B. bronchiseptica in several natural-host infection models. FHA(Bp) was able to substitute for FHA from B. bronchiseptica (FHA(Bb)) with regard to its ability to mediate adherence to several epithelial and macrophage like cell lines in vitro, but it was unable to substitute for FHA(Bb) in vivo. Specifically, FHA(Bb), but not FHA(Bp), allowed B. bronchiseptica to colonize the lower respiratory tracts of rats, to modulate the inflammatory response in the lungs of immunocompetent mice, resulting in decreased lung damage and increased bacterial persistence, to induce a robust anti-Bordetella antibody response in these immunocompetent mice, and to overcome innate immunity and cause a lethal infection in immunodeficient mice. These results indicate a critical role for FHA in B. bronchiseptica-mediated immunomodulation, and they suggest a role for FHA in host specificity. PMID- 16339900 TI - The spread of attention across modalities and space in a multisensory object. AB - Attending to a stimulus is known to enhance the neural responses to that stimulus. Recent experiments on visual attention have shown that this modulation can have object-based characteristics, such that, when certain parts of a visual object are attended, other parts automatically also receive enhanced processing. Here, we investigated whether visual attention can modulate neural responses to other components of a multisensory object defined by synchronous, but spatially disparate, auditory and visual stimuli. The audiovisual integration of such multisensory stimuli typically leads to mislocalization of the sound toward the visual stimulus (ventriloquism illusion). Using event-related potentials and functional MRI, we found that the brain's response to task-irrelevant sounds occurring synchronously with a visual stimulus from a different location was larger when that accompanying visual stimulus was attended versus unattended. The event-related potential effect consisted of sustained, frontally distributed, brain activity that emerged relatively late in processing, an effect resembling attention-related enhancements seen at earlier latencies during intramodal auditory attention. Moreover, the functional MRI data confirmed that the effect included specific enhancement of activity in auditory cortex. These findings indicate that attention to one sensory modality can spread to encompass simultaneous signals from another modality, even when they are task-irrelevant and from a different location. This cross-modal attentional spread appears to reflect an object-based, late selection process wherein spatially discrepant auditory stimulation is grouped with synchronous attended visual input into a multisensory object, resulting in the auditory information being pulled into the attentional spotlight and bestowed with enhanced processing. PMID- 16339901 TI - An antiviral role for the RNA interference machinery in Caenorhabditis elegans. AB - RNA interference (RNAi) is a sequence-specific gene-silencing mechanism triggered by exogenous dsRNA. In plants an RNAi-like mechanism defends against viruses, but the hypothesis that animals possess a similar natural antiviral mechanism related to RNAi remains relatively untested. To test whether genes needed for RNAi defend animal cells against virus infection, we infected wild-type and RNAi-defective cells of the nematode C. elegans with vesicular stomatitis virus engineered to encode a GFP fusion protein. We show that upon infection, cells lacking components of the RNAi apparatus produce more GFP and infective particles than wild-type cells. Furthermore, we show that mutant cells with enhanced RNAi produce less GFP. Our observation that multiple genes required for RNAi are also required for resistance to vesicular stomatitis virus suggests that the RNAi machinery functions in resistance to viruses in nature. PMID- 16339902 TI - Mapping Dmef2-binding regulatory modules by using a ChIP-enriched in silico targets approach. AB - Mapping the regulatory modules to which transcription factors bind in vivo is a key step toward understanding of global gene expression programs. We have developed a chromatin immunoprecipitation (ChIP)-chip strategy for identifying factor-specific regulatory regions acting in vivo. This method, called the ChIP enriched in silico targets (ChEST) approach, combines immunoprecipitation of cross-linked protein-DNA complexes (X-ChIP) with in silico prediction of targets and generation of computed DNA microarrays. We report the use of ChEST in Drosophila to identify several previously unknown targets of myocyte enhancer factor 2 (MEF2), a key regulator of myogenic differentiation. Our approach was validated by demonstrating that the identified sequences act as enhancers in vivo and are able to drive reporter gene expression specifically in MEF2-positive muscle cells. Presented here, the ChEST strategy was originally designed to identify regulatory modules in Drosophila, but it can be adapted for any sequenced and annotated genome. PMID- 16339903 TI - Slow growth rates of Amazonian trees: consequences for carbon cycling. AB - Quantifying age structure and tree growth rate of Amazonian forests is essential for understanding their role in the carbon cycle. Here, we use radiocarbon dating and direct measurement of diameter increment to document unexpectedly slow growth rates for trees from three locations spanning the Brazilian Amazon basin. Central Amazon trees, averaging only approximately 1 mm/year diameter increment, grow half as fast as those from areas with more seasonal rainfall to the east and west. Slow growth rates mean that trees can attain great ages; across our sites we estimate 17-50% of trees with diameter >10 cm have ages exceeding 300 years. Whereas a few emergent trees that make up a large portion of the biomass grow faster, small trees that are more abundant grow slowly and attain ages of hundreds of years. The mean age of carbon in living trees (60-110 years) is within the range of or slightly longer than the mean residence time calculated from C inventory divided by annual C allocation to wood growth (40-100 years). Faster C turnover is observed in stands with overall higher rates of diameter increment and a larger fraction of the biomass in large, fast-growing trees. As a consequence, forests can recover biomass relatively quickly after disturbance, whereas recovering species composition may take many centuries. Carbon cycle models that apply a single turnover time for carbon in forest biomass do not account for variations in life strategy and therefore may overestimate the carbon sequestration potential of Amazon forests. PMID- 16339904 TI - A papain-like enzyme at work: native and acyl-enzyme intermediate structures in phytochelatin synthesis. AB - Phytochelatin synthase (PCS) is a key enzyme for heavy-metal detoxification in plants. PCS catalyzes the production of glutathione (GSH)-derived peptides (called phytochelatins or PCs) that bind heavy-metal ions before vacuolar sequestration. The enzyme can also hydrolyze GSH and GS-conjugated xenobiotics. In the cyanobacterium Nostoc, the enzyme (NsPCS) contains only the catalytic domain of the eukaryotic synthase and can act as a GSH hydrolase and weakly as a peptide ligase. The crystal structure of NsPCS in its native form solved at a 2.0 A resolution shows that NsPCS is a dimer that belongs to the papain superfamily of cysteine proteases, with a conserved catalytic machinery. Moreover, the structure of the protein solved as a complex with GSH at a 1.4-A resolution reveals a gamma-glutamyl cysteine acyl-enzyme intermediate stabilized in a cavity of the protein adjacent to a second putative GSH binding site. GSH hydrolase and PCS activities of the enzyme are discussed in the light of both structures. PMID- 16339905 TI - Interaction of nephrocystin-4 and RPGRIP1 is disrupted by nephronophthisis or Leber congenital amaurosis-associated mutations. AB - RPGR-interacting protein 1 (RPGRIP1) is a key component of cone and rod photoreceptor cells, where it interacts with RPGR (retinitis pigmentosa GTPase regulator). Mutations in RPGRIP1 lead to autosomal recessive congenital blindness [Leber congenital amaurosis (LCA)]. Most LCA-associated missense mutations in RPGRIP1 are located in a segment that encodes two C2 domains. Based on the C2 domain of novel protein kinase C epsilon (PKC epsilon), we built a 3D-homology model for the C-terminal C2 domain of RPGRIP1. This model revealed a potential Ca2+-binding site that was predicted to be disrupted by a missense mutation in RPGRIP1, which was previously identified in an LCA patient. Through yeast two hybrid screening of a retinal cDNA library, we found this C2 domain to specifically bind to nephrocystin-4, encoded by NPHP4. Mutations in NPHP4 are associated with nephronophthisis and a combination of nephronophthisis and retinitis pigmentosa called Senior-Loken syndrome (SLSN). We show that RPGRIP1 and nephrocystin-4 interact strongly in vitro and in vivo, and that they colocalize in the retina, matching the panretinal localization pattern of specific RPGRIP1 isoforms. Their interaction is disrupted by either mutations in RPGRIP1, found in patients with LCA, or by mutations in NPHP4, found in patients with nephronophthisis or SLSN. Thus, we provide evidence for the involvement of this disrupted interaction in the retinal dystrophy of both SLSN and LCA patients. PMID- 16339906 TI - Optical detection of rate-determining ion-modulated conformational changes of the ether-a-go-go K+ channel voltage sensor. AB - In voltage-dependent ether-a-go-go (eag) K+ channels, the process of activation is modulated by Mg2+ and other divalent cations, which bind to a site in the voltage sensor and slow channel opening. Previous analysis of eag ionic and gating currents indicated that Mg2+ has a much larger effect on ionic than gating current kinetics. From this, we hypothesized that ion binding modulates voltage sensor conformational changes that are poorly represented in gating current recordings. We have now tested this proposal by using a combined electrophysiological and optical approach. We find that a fluorescent probe attached near S4 in the voltage sensor reports on two phases of the activation process. One component of the optical signal corresponds to the main charge moving conformational changes of the voltage sensor. This is the phase of activation that is well represented in gating current recordings. Another component of the optical signal reflects voltage sensor conformational changes that occur at more hyperpolarized potentials. These transitions, which are rate determining for activation and highly modulated by Mg2+, have not been detected in gating current recordings. Our results demonstrate that the eag voltage sensor undergoes conformational changes that have gone undetected in electrical measurements. These transitions account for the time course of eag activation in the presence and absence of extracellular Mg2+. PMID- 16339907 TI - Bacillus thuringiensis subsp. israelensis Cyt1Aa synergizes Cry11Aa toxin by functioning as a membrane-bound receptor. AB - Bacillus thuringiensis subsp. israelensis produces crystal proteins, Cry (4Aa, 4Ba, 10Aa, and 11Aa) and Cyt (1Aa and 2Ba) proteins, toxic to mosquito vectors of human diseases. Cyt1Aa overcomes insect resistance to Cry11Aa and Cry4 toxins and synergizes the toxicity of these toxins. However, the molecular mechanism of synergism remains unsolved. Here, we provide evidence that Cyt1Aa functions as a receptor of Cry11Aa. Sequential-binding analysis of Cyt1Aa and Cry11Aa revealed that Cyt1Aa binding to Aedes aegypti brush border membrane vesicles enhanced the binding of biotinylated-Cry11Aa. The Cyt1Aa- and Cry11Aa-binding epitopes were mapped by means of the yeast two-hybrid system, peptide arrays, and heterologous competition assays with synthetic peptides. Two exposed regions in Cyt1Aa, loop beta6-alphaE and part of beta7, bind Cry11Aa. On the other side, Cry11Aa binds Cyt1Aa proteins by means of domain II-loop alpha8 and beta-4, which are also involved in midgut receptor interaction. Characterization of single-point mutations in Cry11Aa and Cyt1Aa revealed key Cry11Aa (S259 and E266) and Cyt1Aa (K198, E204 and K225) residues involved in the interaction of both proteins and in synergism. Additionally, a Cyt1Aa loop beta6-alphaE mutant (K198A) with enhanced synergism to Cry11Aa was isolated. Data provided here strongly indicates that Cyt1Aa synergizes or suppresses resistance to Cry11Aa toxin by functioning as a membrane-bound receptor. Bacillus thuringiensis subsp. israelensis is a highly effective pathogenic bacterium because it produces a toxin and also its functional receptor, promoting toxin binding to the target membrane and causing toxicity. PMID- 16339908 TI - The tethered motor domain of a kinesin-microtubule complex catalyzes reversible synthesis of bound ATP. AB - Although the steps for the forward reaction of ATP hydrolysis by the motor protein kinesin have been studied extensively, the rates for the reverse reactions and thus the energy changes at each step are not as well defined. Oxygen isotopic exchange between water and P(i) was used to evaluate the reverse rates. The fraction of the kinesin x ADP x P(i) complex that reverts to ATP before release of P(i) during net hydrolysis was approximately 0 and approximately 2.6% in the absence and presence of microtubules (MTs), respectively. The rate of synthesis of bound ATP from free P(i) and the MT x kinesin x ADP complex was approximately 1.7 M(-1) x s(-1) (K0.5 ADP = 70 microM) with monomeric kinesin in the absence of net hydrolysis. Synthesis of bound ATP from the ADP of the tethered head of a dimer-MT complex was 20-fold faster than for the monomer-MT complex. This MT-activated ATP synthesis at the tethered head is in marked contrast to the lack of MT stimulation of ADP release from the same site. The more rapid ATP synthesis with dimers suggests that the tethered head binds behind the strongly attached head, because this positions the neck linker of the tethered head toward the plus end of the MT and would thus facilitate its docking on synthesis of ATP. The observed rate of ATP synthesis also puts limits on the overall energetics that suggest that a significant fraction of the free energy of ATP hydrolysis is available to drive the docking of the neck linker on binding of ATP. PMID- 16339910 TI - Shoot circumnutation and winding movements require gravisensing cells. AB - Circumnutation and winding in plants are universal growth movements that allow plants to survive despite their sessile nature. However, the detailed molecular mechanisms controlling these phenomena remain unclear. We previously found that a gravitropic mutant of Japanese morning glory (Pharbitis nil or Ipomoea nil), Shidare-asagao (weeping), is defective not only in circumnutation but also in the winding response. This phenotype is similar to that of the Arabidopsis SCARECROW (SCR) mutant. We therefore investigated whether morning glory SCR (PnSCR) is involved in the weeping phenotype. We found that one amino acid was inserted into the highly conserved VHIID motif in weeping-type PnSCR; this mutation caused abnormal endodermal differentiation. We introduced either the mutant or WT PnSCR into Arabidopsis scr mutants for complementation tests. PnSCR of the WT, but not of weeping, rescued the shoot gravitropism and circumnutation of scr. These results show that both the abnormal gravitropism and the circumnutation defect in weeping are attributable to a loss of PnSCR function. Thus, our data show that gravisensing endodermal cells are indispensable for shoot circumnutation and the winding response and that PnSCR is responsible for the abnormal phenotypes of weeping. PMID- 16339911 TI - A novel, selective, and orally available antagonist for CC chemokine receptor 3. AB - CC chemokine ligand 11 (CCL11/eotaxin) and other CC chemokine receptor 3 (CCR3) ligands (CCL24/eotaxin-2, CCL26/eotaxin-3, CCL13/monocyte chemotactic protein-4, etc.) play important roles in the chemotaxis and activation of eosinophils and other CCR3-expressing cells (basophils, mast cells, and CD4(+) T helper 2 cells) in allergic inflammation incidents, including asthma and rhinitis. A newly synthesized compound, N-{(3R)-1-[(6-fluoro-2-naphthyl)methyl]pyrrolidin-3-yl}-2 {1-[(5-hydroxy-3-methylpyridin-2-yl)carbonyl]piperidin-4-ylidene}-acetamide hemifumarate (YM-355179), inhibited the binding of CCL11 and CCL5/regulated on activation normal T cell expressed and secreted to CCR3-expressing B300-19 cells with IC(50) values of 7.6 and 24 nM, respectively. In contrast, YM-355179 did not affect the binding of CCL5 to CCR1 or CCR5. In functional assays, YM-355179 inhibited CCL11-induced, intracellular Ca(2+) influx, chemotaxis, and eosinophil degranulation with IC(50) values of 8.0, 24, and 29 nM, respectively. YM-355179 did not, however, affect any CC chemokine receptor (CCR1, CCR2, CCR4, or CCR5) mediated Ca(2+) influx signals. Furthermore, oral administration of YM-355179 (1 mg/kg) inhibited CCL11-induced shape change of whole blood eosinophils in cynomolgus monkeys. Intravenous injection of YM-355179 (1 mg/kg) also inhibited eosinophil infiltration into airways of cynomolgus monkeys after segmental bronchoprovocation with CCL11. These results indicate that YM-355179 is a novel, selective, and orally available CCR3 antagonist with therapeutic potential for treating eosinophil-related allergic inflammatory diseases. PMID- 16339909 TI - Neutralizing antibody responses drive the evolution of human immunodeficiency virus type 1 envelope during recent HIV infection. AB - HIV type 1 (HIV-1) can rapidly escape from neutralizing antibody responses. The genetic basis of this escape in vivo is poorly understood. We compared the pattern of evolution of the HIV-1 env gene between individuals with recent HIV infection whose virus exhibited either a low or a high rate of escape from neutralizing antibody responses. We demonstrate that the rate of viral escape at a phenotypic level is highly variable among individuals, and is strongly correlated with the rate of amino acid substitutions. We show that dramatic escape from neutralizing antibodies can occur in the relative absence of changes in glycosylation or insertions and deletions ("indels") in the envelope; conversely, changes in glycosylation and indels occur even in the absence of neutralizing antibody responses. Comparison of our data with the predictions of a mathematical model support a mechanism in which escape from neutralizing antibodies occurs via many amino acid substitutions, with low cross neutralization between closely related viral strains. Our results suggest that autologous neutralizing antibody responses may play a pivotal role in the diversification of HIV-1 envelope during the early stages of infection. PMID- 16339912 TI - Developmental expression of aryl, estrogen, and hydroxysteroid sulfotransferases in pre- and postnatal human liver. AB - Aryl- (SULT1A1), estrogen- (SULT1E1), and hydroxysteroid- (SULT2A1) sulfotransferases (SULTs) are active determinants of xenobiotic detoxication and hormone metabolism in the adult human liver. To investigate the role of these conjugating enzymes in the developing human liver, the ontogeny of immunoreactive SULT1A1, SULT1E1, and SULT2A1 expression was characterized in a series of 235 pre and postnatal human liver cytosols ranging in age from early gestation to a postnatal age of 18 years. Interindividual variability in expression levels was apparent for all three SULTs in pre- and postnatal liver samples. Expression of the three SULTs displayed distinctly different developmental profiles. Semiquantitative Western blot analyses indicated that SULT1A1 and SULT2A1 immunoreactive protein levels were readily detectable in the majority of developmental human liver cytosols throughout the prenatal period. Whereas SULT1A1 expression did not differ significantly among the various developmental stages, SULT2A1 expression increased during the third trimester of gestation and continued to increase during postnatal life. By contrast, SULT1E1, a cardinal estrogen-inactivating enzyme, achieved the highest levels of expression during the earliest periods of gestation in prenatal male livers, indicating a requisite role for estrogen inactivation in the developing male. The present analysis suggests that divergent regulatory mechanisms are responsible for the differential patterns of hepatic SULT1A1, SULT1E1, and SULT2A1 immunoreactive protein levels that occur during pre- and postnatal human development, and implicates a major role for sulfotransferase expression in the developing fetus. PMID- 16339913 TI - Pharmacological characterization of (E)-N-(4-fluorobut-2-enyl)-2beta-carbomethoxy 3beta-(4'-tolyl)nortropane (LBT-999) as a highly promising fluorinated ligand for the dopamine transporter. AB - In the aim to develop an efficient fluorinated probe for positron emission tomography (PET) exploration of the dopamine transporter (DAT), we studied several in vitro and in vivo characteristics of the phenyltropane derivative (E) N-(4-fluorobut-2-enyl)-2beta-carbomethoxy-3beta-(4'-tolyl)nortropane (LBT-999). In vitro on rat striatal membrane, [(3)H]LBT-999 bound to a single site with a K(d) of 9 nM, B(max) of 17 pmol/mg protein, and a very high selectivity for the DAT [IC(50) for 1-{2-[bis-(4-fluorophenyl)-methoxy]ethyl}-4-(3 phenylpropyl)piperazine (GBR 12909) and (E)-N-(3-iodoprop-2-enyl)-2beta carbomethoxy-3beta-(4'-methylphenyl)nortropane (PE2I): 2.4 and 18 nM, respectively; IC(50) for paroxetine, citalopram, N,N-dimethyl-2-(2-amino-4 methylphenyl thio)benzylamine, nisoxetine, and desipramine >1 muM]. In vitro on post-mortem human brain sections, LBT-999 bound with high intensity to the caudate-putamen, weakly to the thalamus, and not in the neocortex and cerebellum. This binding was totally abolished in the presence of PE2I. Ex vivo cerebral biodistribution of [(11)C]LBT-999 in rats showed striatum/cerebellum radioactivity ratios of 18 and 25 at 30 and 60 min postinjection, respectively. This accumulation was strongly prevented by preinjection of GBR 12909, whereas paroxetine and nisoxetine had no effect. An in vivo kinetic PET study in three baboons showed a fast and very high uptake in the striatum, with a plateau at 30 min postinjection and a maximal putamen/cerebellum ratio of 30. Taken together, these findings demonstrate that LBT-999 is a highly promising agent for in vivo exploration of the DAT. This probe is currently labeled with (18)F for further characterizations. PMID- 16339914 TI - Adenosine inhibits tumor necrosis factor-alpha release from mouse peritoneal macrophages via A2A and A2B but not the A3 adenosine receptor. AB - Adenosine is elaborated in injured tissues where it suppresses inflammatory responses of essentially all immune cells, including production of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha). Most of the anti-inflammatory actions of adenosine have been attributed to signaling through the A(2A) adenosine receptor (A(2A)AR). Previously, however, it has been shown that the A(3)AR agonist N(6)-(3-iodobenzyl)adenosine-5'-N-methylcarboxamide (IB-MECA) potently inhibited TNF-alpha release from macrophages obtained from A(2A)AR "knockout" (A(2A)KO) mice, suggesting that the A(3)AR may also regulate cytokine expression. Here, we confirmed that the A(2A)AR is the primary AR subtype that suppresses TNF-alpha release from thioglycollate-elicited mouse peritoneal macrophages induced by both Toll-like receptor-dependent (TLR) and TLR independent stimuli, but we determined that the A(2B)AR rather than the A(3)AR mediates the non-A(2A)AR actions of adenosine since 1) the ability of IB-MECA to inhibit TNF-alpha release was not altered in macrophages isolated from A(3)KO mice, and 2) the A(2B)AR antagonist 1,3-dipropyl-8-[4-[((4 cyanophenyl)carbamoylmethyl)oxy]phenyl]xanthine (MRS 1754) blocked the ability of the nonselective AR agonist adenosine-5'-N-ethylcarboxamide (NECA) to inhibit TNF alpha release from macrophages isolated from A(2A)KO mice. Although A(2B)ARs seem capable of inhibiting TNF-alpha release, the A(2A)AR plays a dominant suppressive role since MRS 1754 did not block the ability of NECA to inhibit TNF-alpha release from macrophages isolated from wild-type (WT) mice. Furthermore, the potency and efficacy of adenosine to inhibit TNF-alpha release from WT macrophages were not influenced by blocking A(2B)ARs with MRS 1754. The data indicate that adenosine suppresses TNF-alpha release from macrophages primarily via A(2A)ARs, although the A(2B)AR seems to play an underlying inhibitory role that may contribute to the anti-inflammatory actions of adenosine under select circumstances. PMID- 16339915 TI - Noninvasive mapping of reactive oxygen species by in vivo electron spin resonance spectroscopy in indomethacin-induced gastric ulcers in rats. AB - Reactive oxygen species (ROS) are thought to be involved in the gastric ulcer formation induced by indomethacin, a typical nonsteroidal anti-inflammatory drug. However, the location and the time course of ROS generation remain unknown. To assess the sites of ROS generation, we applied the noninvasive measurement of ROS to indomethacin-treated rats. By giving orally a membrane-permeable or impermeable probe, the spectra were collected as a function of time by in vivo 300-MHz electron spin resonance (ESR) spectroscopy. The ESR signal-decay rates of membrane-permeable probes, hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) and 3-methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine-1-oxyl, in the gastric mucosal region were significantly enhanced 1 h after indomethacin treatment, and they both caused the protection of ulcer formation; however, membrane-impermeable probes, carboxy- and trimethylammonium-TEMPO, which did not exhibit the enhanced signal decay, had no effect on ulcer formation. The enhanced signal decay in the gastric mucosa was suppressed by coadministration of the antioxidants tiron or dimethylthiourea with the nitroxyl probe. The results suggest that the enhanced signal-decay rates in the gastric ulcers observed by in vivo ESR are associated with protective effects. The enhanced signal decay caused by ROS generation in stomach, contributing to the ulcer formation induced by indomethacin, is also suggested to occur at the gastric mucus layer or the interface or the intracellular compartment of epithelial cells. Overall, these results show the potentials of noninvasive assessment of ROS production and the sites of damage by in vivo ESR using nitroxyl probes directed to specific subcellular regions. PMID- 16339916 TI - Functional human corneal endothelial cell sheets harvested from temperature responsive culture surfaces. AB - This study reports a new method for fabricating bioengineered human corneal endothelial cell sheets suitable for ocular surgery and repair. We have initially cultured human corneal endothelial cells on type IV collagen-coated dishes and, after several passages, expanded cells were then seeded onto novel temperature responsive culture dishes. Four weeks after reaching confluence, these cultured endothelial cells were harvested as intact monolayer cell sheets by simple temperature reduction without enzymatic treatment. Scanning electron microscopy indicated that these cells were primarily hexagonal with numerous microvilli and cilia, similar to the native corneal endothelium. The Na+, K+-ATPase pump sites were located at the cell borders as in vivo. Moreover, cell densities and numbers of pump sites were identical to those of in vivo human corneal endothelium under optimized conditions. A 3H-ouabain binding analysis demonstrated a linear proportionality for cell pump density between confluent cell densities of 575 cells/mm2 and 3070 cells/mm2. We also confirmed Na+, K+-ATPase activity in the sheets in vitro. Xenograft transplantation results showed that the fabricated sheets retain their function of maintaining proper stromal hydration in vivo. We have established a regimen to culture and proliferate human corneal endothelial cells and fabricate endothelial sheets ex vivo morphologically and functionally similar to the native corneal endothelium. Our results support the value of harvested cell sheets for clinical applications in ocular reconstructive surgery in patients with ocular endothelial decompensation. PMID- 16339917 TI - Serotonin transporter polymorphisms in familial and idiopathic pulmonary arterial hypertension. AB - RATIONALE: Serotonin is a pulmonary vasoconstrictor and smooth muscle cell mitogen. The serotonin transporter (SERT) is abundant in pulmonary vascular smooth muscle. Compared with the short (S) allele, the long (L) SERT promoter allele is associated with increased SERT transcription and more severe pulmonary hypertension in a cohort of patients with chronic obstructive pulmonary disease, and was more prevalent in a cohort with idiopathic pulmonary arterial hypertension (IPAH), compared with control subjects. OBJECTIVE: We hypothesized that the SERT L allele would associate with an earlier age at diagnosis and/or shorter survival interval in pulmonary arterial hypertension (PAH) than the S allele. METHODS: SERT promoters from 166 familial PAH (FPAH), 83 IPAH, and 125 control subjects were sequenced. One hundred twenty-seven of the patients with FPAH had a known mutation in bone morphogenetic protein receptor 2 (BMPR2). RESULTS: The mean age at diagnosis was 35.8 yr in patients with FPAH and 41.1 yr in patients with IPAH (p = 0.02). There were no significant differences in distribution of the LL, LS, or SS genotypes in IPAH, FPAH, or unaffected BMPR2 mutation carriers. In FPAH, the LL genotype was associated with an earlier age at diagnosis (p < 0.02). CONCLUSIONS: In patients with IPAH, these SERT genotypes do not correlate with age at diagnosis or survival interval. In patients with FPAH, the LL genotype correlates with an earlier age at diagnosis than SL or SS, although survival among the groups was similar. The correlation of the SERT promoter polymorphism with age at diagnosis in FPAH suggests a possible relationship between the SERT and BMPR2. PMID- 16339918 TI - Progression of airway dysplasia and C-reactive protein in smokers at high risk of lung cancer. AB - RATIONALE: Chronic inflammation has been implicated in the development of airway dysplasia and lung cancer. It is unclear whether circulating biomarkers of inflammation could be used to predict progression of airway dysplasia. OBJECTIVE: We determined whether circulating levels of C-reactive protein (CRP) or other inflammatory biomarkers could predict progression of bronchial dysplasia in smokers over 6 mo. METHODS: The plasma levels of CRP, interleukins 6 and 8, and monocyte chemoattractant protein 1 were measured at baseline in 65 ex- and current smokers who had at least one site of bronchial dysplasia > 1.2 mm in size. Additional bronchial biopsies were taken after 6 mo from the same sites where dysplastic lesions were discovered at baseline. Progressive dysplastic lesions were defined as worsening of the dysplastic lesion by at least two grades or development of new dysplastic lesions. RESULTS: Half of the participants developed progressive dysplastic lesions after 6 mo. The baseline CRP levels in these participants were 64% higher than those without progressive disease (p = 0.027). Only one of eight (13%) participants with CRP < or = 0.5 mg/L developed progressive disease, whereas 31 of 57 (54%) participants with CRP > 0.5 mg/L developed progressive disease (p = 0.011). The odds of developing progressive disease were 9.6-fold higher in the latter than in the former group. CONCLUSION: Plasma CRP, in concert with lung function and pack-years of smoking, appears to have excellent predictive powers in identifying participants with bronchial dyplastic lesions whose lesions progress to more advanced stages of dysplasia. PMID- 16339919 TI - Regulatory T cells are expanded in blood and disease sites in patients with tuberculosis. AB - RATIONALE: T-cell responses during tuberculosis (TB) help contain Mycobacterium tuberculosis in vivo but also cause collateral damage to host tissues. Immune regulatory mechanisms may limit this immunopathology, and suppressed cellular immune responses in patients with TB suggest the presence of regulatory activity. CD4+CD25(high) regulatory T cells mediate suppressed cellular immunity in several chronic infections but have not been described in TB. OBJECTIVE: To determine whether regulatory T cells are increased in patients with TB and whether they suppress cellular immune responses. METHODS: We compared the frequency of circulating regulatory T cells in 27 untreated patients with TB and 23 healthy control subjects using two specific markers: cell-surface CD25 expression and FoxP3 mRNA expression in peripheral blood mononuclear cells. MEASUREMENTS AND MAIN RESULTS: We detected a threefold increase in the frequency of CD4 + CD25(high) T cells (p < 0.001) and a 2.2-fold increase in FoxP3 expression (p = 0.006) in patients with TB, and there was a positive correlation between these markers (r = 0.58, p < 0.001). Increased expression of interleukin-10 and transforming growth factor-beta1 mRNA was also detected in patients with TB but did not correlate with regulatory T-cell markers. Ex vivo depletion of CD4 + CD25(high) cells from peripheral blood mononuclear cells resulted in increased numbers of M. tuberculosis antigen-specific IFN-gamma-producing T cells in seven of eight patients with TB (p = 0.005). Finally, FoxP3 expression was increased 2.3-fold in patients with extrapulmonary TB compared with patients with purely pulmonary TB (p = 0.01) and was amplified 2.6-fold at disease sites relative to blood (p = 0.043). CONCLUSIONS: Regulatory T cells are expanded in patients with TB and may contribute to suppression of Th1-type immune responses. PMID- 16339920 TI - The inflammatory role of platelets in cystic fibrosis. AB - Platelets are an important, albeit generally underappreciated, component of the inflammatory cascade. Platelets are known to contribute to inflammation in atherosclerosis, stroke, and asthma. They produce a large number of proinflammatory lipid mediators and cytokines, and play a vital role in recruitment of leukocytes into inflamed tissue. We review the role of platelets in inflammation, how they assist in the recruitment of leukocytes into lung tissue in asthma, and evidence of their dysfunction in cystic fibrosis (CF). Platelet dysfunction in CF could contribute to pulmonary inflammation and tissue destruction. We hypothesize that platelet activation is important in CF lung disease and suggest research avenues that might help elucidate the role of activated platelets in CF. PMID- 16339921 TI - Titin and diaphragm dysfunction in chronic obstructive pulmonary disease. AB - RATIONALE: Recently, we have shown that Ca2+-activated force generation in diaphragm single fibers is impaired in patients with mild to moderate chronic obstructive pulmonary disease (COPD). For optimal active-force generation, the passive elasticity provided by titin is indispensable. OBJECTIVES: In the present study, we determined the passive-tension-length relations of single fibers of patients with mild to moderate COPD, hypothesizing that passive-elastic properties of diaphragm fibers are compromised. METHODS: Passive-tension-length relations were determined in diaphragm fibers from patients with and without COPD (predicted mean FEV1, 76 and 102%, respectively). In diaphragm homogenates titin expression was studied at the protein level by gel electrophoresis and at the transcript level by using a novel titin exon microarray. RESULTS: Diaphragm fibers from patients with COPD generate less passive tension on stretch. Titin content in the diaphragm did not differ between patients with and without COPD. However, titin exon transcript studies revealed up-regulation of seven exons, which code for spring elements in the elastic segment rich in proline, glutamate, valine, and lysine. Immunofluorescence analysis indicated elevated protein expression of the up-regulated splice variant in the COPD diaphragm. Simulation studies on titin molecules including the amino acids encoded by the seven up regulated exons predicted reduced passive-tension generation on molecule stretch. CONCLUSIONS: Passive-tension generation of diaphragm single fibers is reduced in patients with COPD. Our results suggest that alternative splicing of the titin gene, resulting in increased length of the elastic segment rich in proline, glutamate, valine, and lysine, is involved. Interestingly, these changes occur already in patients with mild to moderate COPD. PMID- 16339922 TI - Do inhaled carbon nanoparticles translocate directly into the circulation in humans? AB - RATIONALE: Increased exposure to particulate air pollution (PM(10)) is a risk factor for death and hospitalization with cardiovascular disease. It has been suggested that the nanoparticulate component of PM(10) is capable of translocating into the circulation with the potential for direct effects on the vasculature. OBJECTIVE: The study's aim was to determine the extent to which inhaled technetium-99m ((99m)Tc)-labeled carbon nanoparticles (Technegas) were able to access the systemic circulation. METHODS AND MAIN RESULTS: Ten healthy volunteers inhaled Technegas and blood samples were taken sequentially over the following 6 h. Technegas particles were 4-20 nm in diameter and aggregated to a median particle diameter of approximately 100 nm. Radioactivity was immediately detected in blood, with levels increasing over 60 min. Thin-layer chromatography of whole blood identified a species that moved with the solvent front, corresponding to unbound (99m)Tc-pertechnetate, which was excreted in urine. There was no evidence of particle-bound (99m)Tc at the origin. gamma Camera images demonstrated high levels of Technegas retention (95.6 +/- 1.7% at 6 h) in the lungs, with no accumulation of radioactivity detected over the liver or spleen. CONCLUSIONS: The majority of (99m)Tc-labeled carbon nanoparticles remain within the lung up to 6 h after inhalation. In contrast to previous published studies, thin-layer chromatography did not support the hypothesis that inhaled Technegas carbon nanoparticles pass directly from the lungs into the systemic circulation. PMID- 16339923 TI - Hexosamines, insulin resistance, and the complications of diabetes: current status. AB - The hexosamine biosynthesis pathway (HBP) is a relatively minor branch of glycolysis. Fructose 6-phosphate is converted to glucosamine 6-phosphate, catalyzed by the first and rate-limiting enzyme glutamine:fructose-6-phosphate amidotransferase (GFAT). The major end product is UDP-N-acetylglucosamine (UDP GlcNAc). Along with other amino sugars generated by HBP, it provides essential building blocks for glycosyl side chains, of proteins and lipids. UDP-GlcNAc regulates flux through HBP by regulating GFAT activity and is the obligatory substrate of O-GlcNAc transferase. The latter is a cytosolic and nuclear enzyme that catalyzes a reversible, posttranslational protein modification, transferring GlcNAc in O-linkage (O-GlcNAc) to specific serine/threonine residues of proteins. The metabolic effects of increased flux through HBP are thought to be mediated by increasing O-GlcNAcylation. Several investigators proposed that HBP functions as a cellular nutrient sensor and plays a role in the development of insulin resistance and the vascular complications of diabetes. Increased flux through HBP is required and sufficient for some of the metabolic effects of sustained, increased glucose flux, which promotes the complications of diabetes, e.g., diminished expression of sarcoplasmic reticulum Ca(2+)-ATPase in cardiomyocytes and induction of TGF-beta and plasminogen activator inhibitor-1 in vascular smooth muscle cells, mesangial cells, and aortic endothelial cells. The mechanism was consistent with enhanced O-GlcNAcylation of certain transcription factors. The role of HBP in the development of insulin resistance has been controversial. There are numerous papers showing a correlation between increased flux through HBP and insulin resistance; however, the causal relationship has not been established. More recent experiments in mice overexpressing GFAT in muscle and adipose tissue or exclusively in fat cells suggest that the latter develop in vivo insulin resistance via cross talk between fat cells and muscle. Although the relationship between HBP and insulin resistance may be quite complex, it clearly deserves further study in concert with its role in the complications of diabetes. PMID- 16339924 TI - Aging attenuates both the regularity and joint synchrony of LH and testosterone secretion in normal men: analyses via a model of graded GnRH receptor blockade. AB - Testosterone (T) secretion declines in the aging male, albeit for unknown reasons. From an ensemble perspective, repeated incremental signaling among gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), and T is required to maintain physiological androgen availability. Pattern-regularity statistics, such as univariate approximate entropy (ApEn) and bivariate cross ApEn, provide specific and sensitive model-free measurement of altered multi pathway control. The present study exploits partial muting of one pathway (GnRH drive) to appraise adaptive regulation of LH and T secretion in young and aging individuals. Analyses comprised 100 paired 18-h LH and T concentration time series obtained in 25 healthy men ages 20-72 yr each administered placebo and three graded doses of a specific GnRH-receptor antagonist. Graded blockade of GnRH drive increased the individual regularity of LH and T secretion and the synchrony of LH-T feedforward and T-LH feedback in the cohort as a whole (P<0.001 for each). However, age markedly attenuated ganirelix-induced enhancement of univariate T orderliness and bivariate LH-T feedback and T-LH feedback synchrony (P 99 % replaced by deoxyuridine (dU). The only organisms known to have similar DNA are the Bacillus subtilis-specific bacteriophages PBS1 and PBS2. N-terminal amino acid sequences of four major structural proteins did not show any similarity to (viral) protein sequences in databases, indicating that close relatives of phiR1-37 have not yet been characterized. Genes for two of the structural proteins, p24 and p46, were identified from the partially sequenced phiR1-37 genome. PMID- 16339955 TI - Mutational analysis of the C-terminal domain of the Rhodobacter sphaeroides response regulator PrrA. AB - The Rhodobacter sphaeroides response regulator PrrA directly activates transcription of genes necessary for energy conservation at low O2 tensions and under anaerobic conditions. It is proposed that PrrA homologues contain a C terminal DNA-binding domain (PrrA-CTD) that lacks significant amino acid sequence similarity to those found in other response regulators. To test this hypothesis, single amino acid substitutions were created at 12 residues in the PrrA-CTD. These mutant PrrA proteins were purified and tested for the ability to be phosphorylated by the low-molecular-mass phosphate donor acetyl phosphate, to activate transcription and to bind promoter DNA. Each mutant PrrA protein accepted phosphate from 32P-labelled acetyl phosphate. At micromolar concentrations of acetyl phosphate-treated wild-type PrrA, a single 20 bp region in the PrrA-dependent cycA P2 promoter was protected from DNase I digestion. Of the mutant PrrA proteins tested, only acetyl phosphate-treated PrrA-N168A and PrrA-I177A protected cycA P2 from DNase I digestion at similar protein concentrations compared to wild-type PrrA. The use of in vitro transcription assays with the PrrA-dependent cycA P2 and puc promoters showed that acetyl phosphate-treated PrrA-N168A produced transcript levels similar to that of wild type PrrA at comparable protein concentrations. Using concentrations of acetyl phosphate-treated PrrA that are saturating for the wild-type protein, PrrA-H170A and PrrA-I177A produced <45 % as much transcript as wild-type PrrA. Under identical conditions, the remaining mutant PrrA proteins produced little or no detectable transcripts from either promoter in vitro. Explanations are presented for why these amino acid side chains in the PrrA-CTD are important for its ability to activate transcription. PMID- 16339956 TI - Characterization and spontaneous mutation of a novel gene, polE, involved in pellicle formation in Acetobacter tropicalis SKU1100. AB - Acetobacter tropicalis SKU1100 produces a pellicle polysaccharide, consisting of galactose, glucose and rhamnose, which attaches to the cell surface. This strain forms two types of colony on agar plates: a rough-surfaced colony (R strain) and a mucoid smooth-surfaced colony (S strain). The R strain forms a pellicle, allowing it to float on the medium surface in static culture, while the S strain does not. The pellicle is an assemblage of cells which are tightly associated with capsular polysaccharides (CPS) on the cell surface. In this study, a gene required for pellicle formation by the R strain was investigated by transposon mutagenesis using Tn10. The resulting mutant, designated Pel-, has a smooth surfaced colony and a defect in pellicle formation, as for the S strain. The mutant produced polysaccharide which was instead secreted into the culture medium as extracellular polysaccharide (EPS). An ORF was identified at the Tn10 insertion site, designated polE, upstream of which polABCD genes were also found. The deduced amino acid sequences of polABCD showed a high level of homology to those of rfbBACD which are involved in dTDP-rhamnose synthesis, whereas polE had a relatively low level of homology to glycosyltransferase. In this study a polB (rfbA) disruptant was also prepared, which lacked both CPS and EPS production. A plasmid harbouring the polE or polB genes could restore pellicle formation in the Pel(-) mutant and S strains, and in the DeltapolB mutant, respectively. Thus both polE and polB are evidently involved in pellicle formation, most likely by anchoring polysaccharide to the cell surface and through the production of dTDP rhamnose, respectively. The Pel- and DeltapolB mutants were unable to grow in static culture and became more sensitive to acetic acid due to the loss of pellicle formation. Additionally, this study identified the mutation sites of several S strains which were spontaneously isolated from the original culture and found them to be concentrated in a sequence of 7 C residues in the coding sequence of polE, with the deletion or addition of a single C nucleotide. PMID- 16339957 TI - Medium- to large-sized xylo-oligosaccharides are responsible for xylanase induction in Prevotella bryantii B14. AB - Experiments were done to define the nature of the xylan-derived induction signal for xylanase activity, and evaluate which xylanase genes among the three known ones (xynA, xynB and xynC) are induced by the presence of xylan in Prevotella bryantii B(1)4. During the later stages of exponential growth on glucose, addition of 0.05 % water-soluble xylan (WS-X) stimulated xylanase formation within 30 min. Xylose, xylobiose, xylotriose, xylotetraose, xylopentaose, arabinose and glucuronic acid all failed to induce the xylanase activity. An acid ethanol-soluble fraction of WS-X (approximate degree of polymerization 30) enhanced the activity significantly, whereas the acid-ethanol-insoluble fraction had no effect, unless first digested by the cloned P. bryantii XynC xylanase. These results indicate that medium- to large-sized xylo-oligosaccharides are responsible for induction. The transcription of all three known xylanase genes from P. bryantii was upregulated coordinately by addition of WS-X. There have been relatively few investigations into the regulation of xylanase activity in bacteria, and it appears to be unique that medium- to large-sized xylo oligosaccharides are responsible for induction. PMID- 16339958 TI - Analysis of differential protein expression during growth states of Ferroplasma strains and insights into electron transport for iron oxidation. AB - To investigate the metabolic biochemistry of iron-oxidizing extreme acidophiles, a proteomic analysis of chemomixotrophic and chemo-organotrophic growth, as well as protein expression in the absence of organic carbon, was carried out in Ferroplasma species. Electron transport chain inhibitor studies, spectrophotometric analysis and proteomic results suggest that oxidation of ferrous iron may be mediated by the blue copper-haem protein sulfocyanin and the derived electron passes to a cbb3 terminal electron acceptor. Despite previous suggestions of a putative carbon dioxide fixation pathway, no up-regulation of proteins typically associated with carbon dioxide fixation was evident during incubation in the absence of organic carbon. Although a lack of known carbon dioxide fixation proteins does not constitute proof, the results suggest that these strains are not autotrophic. Proteins putatively involved in central metabolic pathways, a probable sugar permease and flavoproteins were up-regulated during chemo-organotrophic growth in comparison to the protein complement during chemomixotrophic growth. These results reflect a higher energy demand to be derived from the organic carbon during chemo-organotrophic growth. Proteins with suggested function as central metabolic enzymes were expressed at higher levels during chemomixotrophic growth by Ferroplasma acidiphilum Y(T) compared to 'Ferroplasma acidarmanus' Fer1. This study addresses some of the biochemical and bioenergetic questions fundamental for survival of these organisms in extreme acid-leaching environments. PMID- 16339959 TI - bph genes of the thermophilic PCB degrader, Bacillus sp. JF8: characterization of the divergent ring-hydroxylating dioxygenase and hydrolase genes upstream of the Mn-dependent BphC. AB - Bacillus sp. JF8 is a thermophilic polychlorinated biphenyl (PCB) degrader, which utilizes biphenyl and naphthalene. A thermostable, Mn-dependent 2,3 dihydroxybiphenyl 1,2-dioxygenase, BphC_JF8, has been characterized previously. Upstream of bphC are five ORFs exhibiting low homology with, and a different gene order from, previously characterized bph genes. From the 5' to 3' direction the genes are: a putative regulatory gene (bphR), a hydrolase (bphD), the large and small subunits of a ring-hydroxylating dioxygenase (bphA1A2), and a cis-diol dehydrogenase (bphB). Hybridization studies indicate that the genes are located on a plasmid. Ring-hydroxylating activity of recombinant BphA1A2_JF8 towards biphenyl, PCB, naphthalene and benzene was observed in Escherichia coli cells, with complementation of non-specific ferredoxin and ferredoxin reductase by host cell proteins. PCB degradation by recombinant BphA1A2_JF8 showed that the congener specificity of the recombinant enzyme was similar to Bacillus sp. JF8. BphD_JF8, with an optimum temperature of 85 degrees C, exhibited a narrow substrate preference for 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid. The Arrhenius plot of BphD_JF8 was biphasic, with two characteristic energies of activation and a break point at 47 degrees C. PMID- 16339960 TI - The use of mammalian cortical kidney slices for the study of tubule secretion: a pioneering step toward understanding organic anion transport. PMID- 16339961 TI - Regulation of ROMK (Kir1.1) channels: new mechanisms and aspects. AB - This brief review attempts to provide an overview regarding recent developments in the regulation of ROMK channels. Studies performed in ROMK null mice suggest that ROMK cannot only form hometetramers such as the small-conductance (30-pS) K channels but also construct heterotetramers such as the 70-pS K channel in the thick ascending limb (TAL). The expression of ROMK channels in the plasma membrane is regulated by protein tyrosine kinase (PTK), serum and glucorticoid induced kinase (SGK), and with-no-lysine-kinase 4. PTK is involved in mediating the effect of low K intake on ROMK channel activity. Increases in superoxide anions induced by low dietary K intake are responsible for the stimulation of PTK expression and tyrosine phosphorylation of ROMK channels. Finally, a recent study indicated that ROMK channels can be monoubiquitinated and monoubiquitination regulates the surface expression of ROMK channels. PMID- 16339962 TI - Tight junction biology and kidney dysfunction. AB - The epithelial tight junction (TJ) has three major functions. As a "gate," it serves as a regulatory barrier separating and maintaining biological fluid compartments of different composition. As a "fence," it generates and maintains the apicobasal polarity of cells that form the confluent epithelium. Finally, the TJ proteins form a trafficking and signaling platform that regulates cell growth, proliferation, differentiation, and dedifferentiation. Six examples are selected that illustrate the emerging link between TJ dysfunction and kidney disease. First, the glomerular slit diaphragm (GSD) is evolved, in part, from the TJ and, on maturation, exhibits all three functions of the TJ. GSD dysfunction leads to proteinuria and, in some instances, podocyte dedifferentiation and proliferation. Second, accumulating evidence supports epithelial-mesenchymal transformation (EMT) as a major player in renal fibrosis, the final common pathway that leads to end-stage renal failure. EMT is characterized by a loss of cell-cell contact and apicobasal polarity, which are hallmarks of TJ dysfunction. Third, in autosomal dominant polycystic kidney disease, mutations of the polycystins may disrupt their known interactions with the apical junction complex, of which the TJ is a major component. This can lead to disturbances in epithelial polarity regulation with consequent abnormal tubulogenesis and cyst formation. Fourth, evidence for epithelial barrier and polarity dysregulation in the pathogenesis of ischemic acute renal failure will be summarized. Fifth, the association between mutations of paracellin-1, the first TJ channel identified, and clinical disorders of magnesium and calcium wasting and bovine renal fibrosis will be used to highlight an integral TJ protein that can serve multiple TJ functions. Finally, the role of WNK4 protein kinase in shunting chloride across the TJ of the distal nephron will be addressed. PMID- 16339963 TI - Evidence that inhibition of tubular cell apoptosis protects against renal damage and development of fibrosis following ureteric obstruction. AB - Ureteric obstruction is frequently encountered in primary care urology and can lead to damage to the ipsilateral kidney. Relief of all types of obstruction generally leads to the normalization of any deterioration in renal function noted at diagnosis. However, some evidence from animal models suggests that obstruction can cause progressive deleterious effects on renal function and blood pressure control, especially in the presence of preexisting pathologies such as essential hypertension. The last 10 years have seen a proliferation of studies in rodents wherein complete unilateral ureteric obstruction has been used as a model of renal fibrosis. However, the relevance of the findings to human obstructive uropathy has, in many cases, not been the primary aim. In this review, we outline the major events linking damage to the renal parenchyma and cell death to the evolution of fibrosis following obstruction. Special focus is given to the role of apoptosis as a major cause of cell death during and post-complete ureteric obstruction. Several interventions that reduce tubular apoptosis are discussed in terms of their ability to prevent subsequent progression to end-organ damage and fibrosis. PMID- 16339964 TI - Protein kinase B/Akt at a glance. PMID- 16339965 TI - CAR might provide a survival signal for myocardial cells. PMID- 16339966 TI - Host and parasite-derived IKK activities direct distinct temporal phases of NF kappaB activation and target gene expression following Toxoplasma gondii infection. AB - Activation of NF-kappaB by the intracellular pathogen Toxoplasma gondii is associated with the localization of phosphorylated IkappaB alpha to the parasitophorous vacuole membrane (PVM). This is mediated by a parasite-derived IkappaB kinase (TgIKK) activity and is independent of host IKK function. In the present study, we examined the roles of host IKK and parasite-derived TgIKK on the temporal modulation of NF-kappaB activation. Despite the presence of TgIKK activity at the PVM, nuclear translocation of NF-kappaB and subsequent gene expression exhibited a requirement for the host IKK complex. A detailed kinetic analysis of NF-kappaB activation revealed a biphasic, hierarchical and temporally regulated response. We propose a novel paradigm for the modulation of NF-kappaB dependent gene expression by T. gondii that involves both the host IKK complex and TgIKK activity at different phases of infection. Thus, T. gondii effectively alters gene expression in a temporal dimension by exploiting the NF-kappaB signaling machinery and subsequently rewiring the activation circuits of the infected host cell. PMID- 16339967 TI - LEM2 is a novel MAN1-related inner nuclear membrane protein associated with A type lamins. AB - The LEM (lamina-associated polypeptide-emerin-MAN1) domain is a motif shared by a group of lamin-interacting proteins in the inner nuclear membrane (INM) and in the nucleoplasm. The LEM domain mediates binding to a DNA-crosslinking protein, barrier-to-autointegration factor (BAF). We describe a novel, ubiquitously expressed LEM domain protein, LEM2, which is structurally related to MAN1. LEM2 contains an N-terminal LEM motif, two predicted transmembrane domains and a MAN1 Src1p C-terminal (MSC) domain highly homologous to MAN1, but lacks the MAN1 specific C-terminal RNA-recognition motif. Immunofluorescence microscopy of digitonin-treated cells and subcellular fractionation identified LEM2 as a lamina associated protein residing in the INM. LEM2 binds to the lamin C tail in vitro. Targeting of LEM2 to the nuclear envelope requires A-type lamins and is mediated by the N-terminal and transmembrane domains. Highly overexpressed LEM2 accumulates in patches at the nuclear envelope and forms membrane bridges between nuclei of adjacent cells. LEM2 structures recruit A-type lamins, emerin, MAN1 and BAF, whereas lamin B and lamin B receptor are excluded. Our data identify LEM2 as a novel A-type-lamin-associated INM protein involved in nuclear structure organization. PMID- 16339968 TI - BMP2 and FGF2 cooperate to induce neural-crest-like fates from fetal and adult CNS stem cells. AB - CNS stem cells are best characterized by their ability to self-renew and to generate multiple differentiated derivatives, but the effect of mitogenic signals, such as fibroblast growth factor 2 (FGF2), on the positional identity of these cells is not well understood. Here, we report that bone morphogenetic protein 2 (BMP2) induces telencephalic CNS stem cells to fates characteristic of neural crest and choroid plexus mesenchyme, a cell type of undetermined lineage in rodents. This induction occurs both in dissociated cell culture and cortical explants of embryonic day 14.5 (E14.5) embryos, but only when cells have been exposed to FGF2. Neither EGF nor IGF1 can substitute for FGF2. An early step in this response is activation of beta-catenin, a mediator of Wnt activity. The CNS stem cells first undergo an epithelial-to-mesenchymal transition and subsequently differentiate to smooth-muscle and non-CNS glia cells. Similar responses are seen with stem cells from E14.5 cortex, E18.5 cortex and adult subventricular zone, but with a progressive shift toward gliogenesis that is characteristic of normal development. These data indicate that FGF2 confers competence for dorsalization independently of its mitogenic action. This rapid and efficient induction of dorsal fates may allow identification of positional identity effectors that are co-regulated by FGF2 and BMP2. PMID- 16339969 TI - Efficient suppression of FGF-2-induced ERK activation by the cooperative interaction among mammalian Sprouty isoforms. AB - Strict regulation of the receptor tyrosine kinase (RTK)/extracellular signal regulated kinase (ERK) pathway is essential for maintaining balanced growth in multi-cellular organisms. Several negative regulators of the pathway have been identified which include Sprouty proteins. Mammalian cells express four Sprouty isoforms (Sprouty1-4) in an ERK-dependent manner. In this study, we have examined the molecular mechanisms by which Sprouty proteins elicit their inhibitory effects on the RTK/ERK pathway, with special focus on the co-operation among Sprouty isoforms. The four mammalian Sprouty isoforms interact with each other, most probably to form hetero- as well as homo-oligomers through their C-terminal domains. Sprouty1 specifically interacts with Grb2, whereas Sprouty4 interacts with Sos1. Although any of the Sprouty isoforms by itself inhibits the fibroblast growth factor-2 (FGF-2)-induced activation of the ERK pathway significantly, hetero-oligomers show a more pronounced inhibitory activity. The hetero-oligomer formed between Sprouty1 and Sprouty4 exhibits the most potent inhibitory effect on ERK activation through its highly effective ability to suppress the association of Grb2-Sos1 complex with FRS2. The cooperative interactions observed among Sprouty isoforms could represent an advanced system that functions to regulate strictly the activation state of the RTK/ERK pathway in mammalian cells. PMID- 16339970 TI - Assembly of additional heterochromatin distinct from centromere-kinetochore chromatin is required for de novo formation of human artificial chromosome. AB - Alpha-satellite (alphoid) DNA is necessary for de novo formation of human artificial chromosomes (HACs) in human cultured cells. To investigate the relationship among centromeric, transcriptionally permissive and non-permissive chromatin assemblies on de novo HAC formation, we constructed bacterial artificial chromosome (BAC)-based linear HAC vectors whose left vector arms are occupied by beta geo coding genes with or without a functional promoter in addition to a common marker gene on the right arm. Although HACs were successfully generated from the vectors with promoter-less constructs on the left arm in HT1080 cells, we failed to generate a stable HAC from the vectors with a functional promoter on the left arm. Despite this failure in HAC formation, centromere components (CENP-A, CENP-B and CENP-C) assembled at the integration sites correlating with a transcriptionally active state of both marker genes on the vector arms. However, on the stable HAC, chromatin immunoprecipitation analysis showed that HP1alpha and trimethyl histone H3-K9 were enriched at the non-transcribing left vector arm. A transcriptionally active state on both vector arms is not compatible with heterochromatin formation on the introduced BAC DNA, suggesting that epigenetic assembly of heterochromatin is distinct from centromere chromatin assembly and is required for the establishment of a stable artificial chromosome. PMID- 16339971 TI - Incautious reasoning as a pathogenetic factor for the development of psychotic symptoms in schizophrenia. AB - Previous studies indicate that schizophrenia patients draw decisions more hastily than controls. The aim of the present study was to obtain convergent evidence with a new paradigm, designed after the Who Wants to Be a Millionaire television game show. Thirty-two schizophrenia patients and 38 healthy subjects were administered 20 knowledge questions, along with 4 response alternatives. Participants were required to provide probability estimates for each alternative. Whenever a subject was confident that one of the alternatives was correct or was wrong, the subject was asked to indicate this via a decision or rejection rating. Thus, probability estimates and decisions were independently assessed, allowing determination of the point at which probability estimates translate into decisions. Patients and controls gave comparable probability estimates for all alternatives. However, patients committed more erroneous responses, owing to their making decisions in the face of low subjective probability ratings and rejecting alternatives despite rather high probability ratings. The results provide further evidence for the claim that schizophrenia patients make strong judgments based on little information. We propose that a lowered threshold for accepting alternatives provides a parsimonious explanation for the data-gathering bias reported in the literature. PMID- 16339972 TI - Kurt Snyder's personal experience with schizophrenia. PMID- 16339974 TI - Ministers provide updates on avian influenza. PMID- 16339978 TI - Footrot and interdigital dermatitis in sheep: farmers' practices, opinions and attitudes. AB - In 1999, a study was initiated to improve the treatment and control of footrot and interdigital dermatitis in sheep flocks in England and Wales. In November 2000, a retrospective cross-sectional study was conducted in which 392 sheep farmers were asked to estimate the prevalence of footrot and interdigital dermatitis in their flock in the previous 12 months, whether they considered these diseases to be a problem, how they treated and controlled them and their opinion on the success of the treatment and control measures that they used; 209 of them provided usable responses. The farmers tended to be more concerned as the prevalence of the diseases increased; 91 per cent of the farmers with a prevalence of footrot of less than 5 per cent considered it a small or very small problem on their farm, but 51 per cent of the farmers with a prevalence of 5 per cent or more also considered it to be a small or very small problem. Approximately 60 per cent of the farmers who used parenteral antibiotics considered that they were good or excellent at treating footrot, and this treatment was associated with a prevalence of less than 5 per cent. A similar proportion of farmers also considered topical foot sprays and footbathing to be good or excellent for controlling footrot or interdigital dermatitis, but these treatments were not associated with a lower prevalence of footrot or interdigital dermatitis. Of the 29 farmers who used a footrot vaccine, 20 (69 per cent) considered it good to excellent and this was associated with a prevalence of footrot of less than 5 per cent in their flock; however, vaccination was not associated with lower levels of footrot across the whole sample. Farmers spent approximately 34 minutes per sheep per year treating and controlling footrot; 31 per cent were prepared to spend more money and more time to manage footrot, 27 per cent were prepared to spend more time, 19 per cent were prepared to spend more money and 23 per cent were not prepared to do either. The farmers who were willing to invest more money or more money and time had a higher prevalence of footrot. PMID- 16339979 TI - Target-controlled infusion of propofol in dogs--evaluation of four targets for induction of anaesthesia. AB - Four groups of 20 dogs were anaesthetised by means of target-controlled infusions of propofol designed to achieve 2.5 microg/ml, 3.0 microg/ml, 3.5 microg/ml or 4.0 microg/ml of propofol in blood. The dogs' pulse rate and respiratory rate were recorded before premedication and induction, immediately after endotracheal intubation and three and five minutes later (times 0, 3 and 5, respectively), and their arterial blood pressure was recorded oscillometrically just before induction and at times 0, 3 and 5. The targets of 2.5, 3.0, 3.5 and 4.0 microg/ml resulted in the successful induction of anaesthesia in 13 (65 per cent), 16 (80 per cent), 20 (100 per cent) and 20 (100 per cent) of the dogs, respectively. The incidence of postinduction apnoea was 0 (0 per cent), one (5 per cent), two (10 per cent) and eight (40 per cent) at time 5 for groups 2.5, 3.0, 3.5 and 4.0 mug/ml, respectively, and its incidence at time 5 was significantly higher in the 4.0 microg/ml group (P<0.05) than in the other groups. In all the groups there was a significant (P<0.05) decrease in blood pressure between just before induction and the later measurements. Although there were no statistically significant differences between the groups in terms of inducing anaesthesia at a specific target, a target of 3.5 microg/ml appears to ensure a successful induction of anaesthesia without a significant increase in the incidence of apnoea. PMID- 16339980 TI - Protection of pregnant mice against placental and splenic infection by three strains of Chlamydophila abortus with a live 1B vaccine. AB - The efficacy of a live 1B vaccine against three strains of Chlamydophila abortus, AB16, LLG and POS, was assessed in pregnant mice in terms of the reduction in the levels of infection recorded in their placentas, fetuses and spleens. The vaccine was more effective against the AB16 strain than against the LLG and POS strains, suggesting that there are antigenic differences between the three strains. PMID- 16339981 TI - Lawsonia intracellularis infection in a 12-month-old colt in Belgium. PMID- 16339982 TI - Novel type of haemorrhagic hepatosis in guinea fowl. PMID- 16339983 TI - Outbreak of salmonellosis in calves in a dairy herd caused by monophasic Salmonella serovar 9,12:l,v:-. PMID- 16339984 TI - Link between feed bins and BSE cases born after July 1996. PMID- 16339985 TI - Re-emerging risk of hydatid disease in Wales. PMID- 16339986 TI - Quality of life. PMID- 16339987 TI - Animal and human health. PMID- 16339988 TI - Non-therapeutic docking of dogs' tails. PMID- 16339989 TI - Non-therapeutic docking of dogs' tails. PMID- 16339990 TI - Naturally occurring antibodies in rabbit antisera to Pseudomonas aeruginosa. PMID- 16339991 TI - Cytological versus histopathological diagnosis in canine osteosarcoma. PMID- 16339992 TI - Microwave-assisted protein preparation and enzymatic digestion in proteomics. AB - The combinations of gel electrophoresis or LC and mass spectrometry are two popular approaches for large scale protein identification. However, the throughput of both approaches is limited by the speed of the protein digestion process. Present research into fast protein enzymatic digestion has been focused mainly on known proteins, and it is unclear whether these results can be extrapolated to complex protein mixtures. In this study microwave technology was used to develop a fast protein preparation and enzymatic digestion method for protein mixtures. The protein mixtures in solution or in gel were prepared and digested by microwave-assisted protein enzymatic digestion, which rapidly produces peptide fragments. The peptide fragments were further analyzed by capillary LC and ESI-ion trap-MS or MALDI-TOF-MS. The technique was optimized using bovine serum albumin and then applied to human urinary proteins and yeast lysate. The method enabled preparation and digestion of protein mixtures in solution (human urinary proteins) or in gel (yeast lysate) in 6 or 25 min, respectively. Equivalent (in-solution) or better (in-gel) digestion efficiency was obtained using microwave-assisted protein enzymatic digestion compared with the standard overnight digestion method. This new application of microwave technology to protein mixture preparation and enzymatic digestion will hasten the application of proteomic techniques to biological and clinical research. PMID- 16339993 TI - An efficient method for the derivation of mouse embryonic stem cells. AB - Mouse embryonic stem cells (mESCs) represent a unique tool for many researchers; however, the process of ESC derivation is often very inefficient and requires high specialization, training, and expertise. To circumvent these limitations, we aimed to develop a simple and efficient protocol based on the use of commercially available products. Here, we present an optimized protocol that we successfully applied to derive ESCs from several knockout mouse strains (Wnt-1, Wnt-5a, Lrp6, and parkin) with 50%-75% efficiency. The methodology is based on the use of mouse embryonic fibroblast feeders, knockout serum replacement (SR), and minimal handling of the blastocyst. In this protocol, all centrifugation steps (as well as the use of trypsin inhibitor) were avoided and replaced by an ESC medium containing fetal calf serum (FCS) after the trypsinizations. We define the potential advantages and disadvantages of using SR and FCS in individual steps of the protocol. We also characterize the ESCs for the expression of ESC markers by immunohistochemistry, Western blot, and a stem cell focused microarray. In summary, we provide a simplified and improved protocol to derive mESCs that can be useful for laboratories aiming to isolate transgenic mESCs for the first time. PMID- 16339994 TI - Embryonic stem-derived versus somatic neural stem cells: a comparative analysis of their developmental potential and molecular phenotype. AB - Reliable procedures to induce neural commitment of totipotent undifferentiated embryonic stem (ES) cells have provided new tools for investigating the molecular mechanisms underlying cell fate choices. We extensively characterized the developmental potential of ES-induced neural cells obtained using an adaptation of the multistep induction protocol. We provided evidence that ES-derived neural proliferating cells are endowed with stem cell properties such as extensive self renewal capacity and single-cell multipotency. In differentiating conditions, cells matured exclusively into neurons, astrocytes, and oligodendrocytes. All these features have been previously described in only somatic neural stem cells (NSCs). Therefore, we consider it more appropriate to rename our cells ES-derived NSCs. These similarities between the two NSC populations induced us to carefully compare their proliferation ability and differentiation potential. Although they were very similar in overall behavior, we scored specific differences. For instance, ES-derived NSCs proliferated at higher rate and consistently generated a higher number of neurons compared with somatic NSCs. To further investigate their relationships, we carried out a molecular analysis comparing their transcriptional profiles during proliferation. We observed a large fraction of shared expressed transcripts, including genes previously described to be critical in defining somatic NSC traits. Among the genes differently expressed, candidate genes possibly responsible for divergences between the two cell types were selected and further investigated. In particular, we showed that an enhanced MAPK (mitogen-activated protein kinase) signaling is acting in ES-induced NSCs, probably triggered by insulin-like growth factor-II. This may contribute to the high proliferation rate exhibited by these cells in culture. PMID- 16339995 TI - In vitro identification of a cord blood CD133+CD34-Lin+ cell subset that gives rise to myeloid dendritic precursors. PMID- 16339996 TI - In vitro modeling of paraxial and lateral mesoderm differentiation reveals early reversibility. AB - Endothelial cells (ECs) are thought to be derived mainly from the vascular endothelial growth factor receptor 2 (VEGFR-2)+ lateral mesoderm during early embryogenesis. In this study, we specified several pathways for EC differentiation using a murine embryonic stem (ES) cell differentiation culture system that is a model for cellular processes during early embryogenesis. Based on the results of in vitro fate analysis, we show that, in the main pathway, committed ECs are differentiated through the VEGFR-2+ platelet-derived growth factor receptor alpha (PDGFR-alpha)- single-positive (VSP) population that is derived from the VEGFR-2+ PDGFR-alpha+ double-positive (DP) population. This major differentiation course was also confirmed using DNA microarray analysis. In addition to this main pathway, however, ECs also can be generated from the VEGFR 2- PDGFR-alpha+ single-positive (PSP) population, which represents the paraxial mesodermal lineage and is also derived from the DP population. Our results strongly suggest that, even after differentiation from the common progenitor DP population into the VSP and PSP populations, these two populations continue spontaneous switching of their surface phenotype, which results in switching of their eventual fates. The rate of this interlineage conversion between VSP and PSP is unexpectedly high. Because of this potential to undergo fate switch, we conclude that ECs can be generated via multiple pathways in in vitro ES cell differentiation. PMID- 16339997 TI - Efficient lentiviral transduction and improved engraftment of human bone marrow mesenchymal cells. AB - Human bone marrow (BM) mesenchymal stem/progenitor cells are potentially attractive targets for ex vivo gene therapy. The potential of lentiviral vectors for transducing BM mesenchymal cells was examined using a self-inactivating vector that expressed the green fluorescent protein (GFP) from an internal cytomegalovirus (CMV) promoter. This vector was compared with oncoretroviral vectors expressing GFP from the CMV promoter or a modified long-terminal repeat that had been optimized for long-term expression in stem cells. The percentage of GFP-positive cells was consistently higher following lentiviral versus oncoretroviral transduction, consistent with increased GFP mRNA levels and increased gene transfer efficiency measured by polymerase chain reaction and Southern blot analysis. In vitro GFP and FVIII expression lasted for several months post-transduction, although expression slowly declined. The transduced cells retained their stem/progenitor cell properties since they were still capable of differentiating along adipogenic and osteogenic lineages in vitro while maintaining high GFP and FVIII expression levels. Implantation of lentivirally transduced human BM mesenchymal cells using collagen scaffolds into immunodeficient mice resulted in efficient engraftment of gene-engineered cells and long-term transgene expression in vivo. These biocompatible BM mesenchymal implants represent a reversible, safe, and versatile protein delivery approach because they can be retrieved in the event of an unexpected adverse reaction or when expression of the protein of interest is no longer required. In conclusion, efficient gene delivery with lentiviral vectors in conjunction with the use of bioengineered reversible scaffolds improves the therapeutic prospects of this novel approach for gene therapy, protein delivery, or tissue engineering. PMID- 16339998 TI - Cholinergic receptor and cyclic stretch-mediated inflammatory gene expression in intact ASM. AB - We tested the hypothesis that cholinergic stimulation and cyclic stretch regulate inflammatory gene expression in intact airway smooth muscle by measuring mRNA expression in bovine tracheal smooth muscle using limited microarray analysis and RT-PCR. Carbachol (1 microM) induced significant increases in the expression of cyclooxygenase (COX)-1, COX-2, IL-8, and plasminogen activator, urokinase type (PLAU) to levels ranging from 1.3- to 3.1-fold of control. Sinusoidal length oscillation at an amplitude of 10% muscle length and a frequency of 1 Hz induced significant increases in the expression of CCL-2, COX-2, IL-1 beta, and IL-6 to levels ranging from 12- to 206-fold of control. Decreasing the oscillatory amplitude by 50% did not significantly change inflammatory gene expression. In contrast, decreasing the oscillatory frequency by 50% significantly attenuated inflammatory gene expression by 76-93%. Nifedipine (1 microM) had an insignificant effect on carbachol-induced gene expression, but significantly inhibited sinusoidal length oscillation-induced inflammatory gene expression by 40-78%. Correlation analysis revealed two groups of genes with differential responses to sinusoidal length oscillation. The highly responsive group included COX-2, IL-6, and IL-8, which exhibited 45- to 364-fold increases in gene expression in response to sinusoidal length oscillation. The moderately responsive group included CCL2 and PLAU, which exhibited 13- to 19-fold increases in gene expression in response to sinusoidal oscillation. These findings suggest that cyclic stretch regulates inflammatory gene expression in intact airway smooth muscle in an amplitude- and frequency-dependent manner by modulating the activity of L-type voltage-gated calcium channels. PMID- 16339999 TI - Syk activation in dendritic cells is essential for airway hyperresponsiveness and inflammation. AB - We evaluated the role of Syk, using an inhibitor, on allergen-induced airway hyperresponsiveness (AHR) and airway inflammation in a system shown to be B cell- and mast cell-independent. Sensitization of BALB/c mice with ovalbumin (OVA) and alum after three consecutive OVA challenges resulted in AHR to inhaled methacholine and airway inflammation. The Syk inhibitor R406 (30 mg/kg, administered orally, twice daily) prevented the development of AHR, increases in eosinophils and lymphocytes and IL-13 levels in bronchoalveolar lavage (BAL) fluid, and goblet cell metaplasia when administered after sensitization and before challenge with OVA. Levels of IL-4, IL-5, and IFN-gamma in BAL fluid and allergen-specific antibody levels in serum were not affected by treatment. Because many of these responses may be influenced by dendritic cell function, we investigated the effect of R406 on bone marrow-derived dendritic cell (BMDC) function. Co-culture of BMDC with immune complexes of OVA and IgG anti-OVA together with OVA-sensitized spleen mononuclear cells resulted in increases in IL 13 production. IL-13 production was inhibited if the BMDCs were pretreated with the Syk inhibitor. Intratracheal transfer of immune complex-pulsed BMDCs (but not nonpulsed BMDCs) to naive mice before airway allergen challenge induced the development of AHR and increases in BAL eosinophils and lymphocytes. All of these responses were inhibited if the transferred BMDCs were pretreated with R406. These results demonstrate that Syk inhibition prevents allergen-induced AHR and airway inflammation after systemic sensitization and challenge, at least in part through alteration of DC function. PMID- 16340000 TI - Apoptosis of airway epithelial cells: human serum sensitive induction by the cathelicidin LL-37. AB - LL-37 is a human cationic host defense peptide that is present in the specific granules of neutrophils, produced by epithelial cells from a variety of tissues, and is upregulated during inflammation, infection, and injury. It has been proposed to have a variety of antimicrobial functions, including both direct antimicrobial activity and immunomodulatory functions. Using the TUNEL assay it was demonstrated that LL-37 induced apoptosis in vitro in the A549 human lung and 16 HBE4o- human airway epithelial cell lines, and in vivo in the murine airway. Peptide-induced apoptosis in vitro involved the activation of caspase pathways and was substantially inhibited by an inhibitor of caspase 3. Apoptosis was also inhibited by human serum, but not fetal bovine serum. Similarly, human but not fetal bovine serum inhibited the cellular internalization of LL-37 and the production of IL-8 in response to LL-37 treatment of epithelial cells. The protective effects of human serum were also observed with high-density lipoproteins but not by the core peptide apolipoprotein A1, providing one possible mechanism of human serum inhibition of apoptosis. We propose that LL-37 induced apoptosis of epithelial cells at low serum tissue sites may have a protective role against bacterial infection. PMID- 16340001 TI - Effects of iron status on transpulmonary transport and tissue distribution of Mn and Fe. AB - Manganese transport into the blood can result from inhaling metal-containing particles. Intestinal manganese and iron absorption is mediated by divalent metal transporter 1 (DMT1) and is upregulated in iron deficiency. Since iron status alters absorption of Fe and Mn in the gut, we tested the hypothesis that iron status may alter pulmonary transport of these metals. DMT1 expression in the lungs was evaluated to explore its role in metal transport. The pharmacokinetics of intratracheally instilled 54Mn or 59Fe in repeatedly bled or iron oxide exposed rats were compared with controls. Iron oxide exposure caused a reduction in pulmonary transport of 54Mn and 59Fe, and decreased uptake in other major organs. Low iron status from repeated bleeding also reduced pulmonary transport of iron but not of manganese. However, uptake of manganese in the brain and of iron in the spleen increased in bled rats. DMT1 transcripts were detected in airway epithelium, alveolar macrophages, and bronchial-associated lymphoid tissue in all rats. Focal increases were seen in particle-containing macrophages and adjacent epithelial cells, but no change was observed in bled rats. Although lung DMT1 expression did not correlate with iron status, differences in pharmacokinetics of instilled metals suggest that their potential toxicity can be modified by iron status. PMID- 16340002 TI - beta2-Adrenoceptor agonist modulates endothelin-1 receptors in human isolated bronchi. AB - Chronic exposure of human isolated bronchi to beta(2)-adrenergic agonists, especially fenoterol, potentiates smooth muscle contraction in response to endothelin-1 (ET-1), a peptide implicated in chronic inflammatory airway diseases. Our objective was to determine whether ET-1 receptors ETA and ETB are involved in fenoterol enhancement. Twenty-two human bronchi were sensitized to ET 1 by prolonged incubation with 0.1 microM fenoterol (15 h, 21 degrees C). Removing the epithelium after fenoterol incubation limited the maximal contraction (0.10+/-0.36 g without epithelium versus 1.18+/-0.22 with, n=8, P=0.04). After 15 h incubation, 14 and 8 paired rings were fixed, respectively, for immunolabeling of bronchial ETA and ETB receptors, and to determine the mRNA expression levels using real-time quantitative reverse transcription polymerase chain reaction. ETA and ETB receptor mRNA expressions were 1.27- +/- 0.14-fold (not significant) and 2.24- +/- 0.28-fold (P<0.01) higher, respectively, in fenoterol-treated bronchi than in paired controls. Fenoterol incubation significantly increased epithelial ETA and ETB receptor labeling intensity scores (P=0.001 and P=0.002, respectively, versus controls), and enhanced the diffuse localization of ETA receptors on the epithelial cells (P=0.002 versus controls), but did not change the ETB-receptor immunolabeling intensity on airway smooth muscle. We conclude that fenoterol-induced sensitization of human isolated bronchi involves epithelial ETA and ETB receptors, which suggests perturbation of the epithelial regulation of airway smooth muscle contraction in response to ET 1. PMID- 16340003 TI - Mechanisms of Chlamydophila pneumoniae-mediated GM-CSF release in human bronchial epithelial cells. AB - Chlamydophila pneumoniae is an important respiratory pathogen. In this study we characterized C. pneumoniae strain TW183-mediated activation of human small airway epithelial cells (SAEC) and the bronchial epithelial cell line BEAS-2B and demonstrated time-dependent secretion of granulocyte macrophage colony stimulating factor (GM-CSF) upon stimulation. TW183 activated p38 mitogen activated protein kinase (MAPK) in epithelial cells. Kinase inhibition by SB202190 blocked Chlamydia-mediated GM-CSF release on mRNA and protein levels. In addition, the chemical inhibitor as well as dominant-negative mutants of p38 MAPK isoforms p38alpha, beta2, and gamma inhibited C. pneumoniae-related NF-kappaB activation. In contrast, blocking of MAPK ERK, c-Jun kinase/JNK, or PI-3 Kinase showed no effect on Chlamydia-related epithelial cell GM-CSF release. Ultraviolet inactivated pathogens as compared with viable bacteria induced a smaller GM-CSF release, suggesting that viable Chlamydiae were only partly required for a full effect. Presence of an antichlamydial outer membrane protein-A (OmpA) antibody reduced and addition of recombinant heat-shock protein 60 from C. pneumoniae (cHsp60, GroEL-1)-enhanced GM-CSF release, suggesting a role of these proteins in epithelial cell activation. Our data demonstrate that C. pneumoniae triggers an early proinflammatory signaling cascade involving p38 MAPK-dependent NF-kappaB activation, resulting in subsequent GM-CSF release. C. pneumoniae-induced epithelial cytokine liberation may contribute significantly to inflammatory airway diseases like chronic obstructive pulmonary disease (COPD) or bronchial asthma. PMID- 16340004 TI - New approaches to the analysis of palindromic sequences from the human genome: evolution and polymorphism of an intronic site at the NF1 locus. AB - The nature of any long palindrome that might exist in the human genome is obscured by the instability of such sequences once cloned in Escherichia coli. We describe and validate a practical alternative to the analysis of naturally occurring palindromes based upon cloning and propagation in Saccharomyces cerevisiae. With this approach we have investigated an intronic sequence in the human Neurofibromatosis 1 (NF1) locus that is represented by multiple conflicting versions in GenBank. We find that the site is highly polymorphic, exhibiting different degrees of palindromy in different individuals. A side-by-side comparison of the same plasmids in E.coli versus. S.cerevisiae demonstrated that the more palindromic alleles were inevitably corrupted upon cloning in E.coli, but could be propagated intact in yeast. The high quality sequence obtained from the yeast-based approach provides insight into the various mechanisms that destabilize a palindrome in E.coli, yeast and humans, into the diversification of a highly polymorphic site within the NF1 locus during primate evolution, and into the association between palindromy and chromosomal translocation. PMID- 16340005 TI - Rapid and quantitative detection of homologous and non-homologous recombination events using three oligonucleotide MLPA. AB - Embryonic stem (ES) cell technology allows modification of the mouse germline from large deletions and insertions to single nucleotide substitutions by homologous recombination. Identification of these rare events demands an accurate and fast detection method. Current methods for detection rely on Southern blotting and/or conventional PCR. Both the techniques have major drawbacks, Southern blotting is time-consuming and PCR can generate false positives. As an alternative, we here demonstrate a novel approach of Multiplex Ligation-dependent Probe Amplification (MLPA) as a quick, quantitative and reliable method for the detection of homologous, non-homologous and incomplete recombination events in ES cell clones. We have adapted MLPA to detect homologous recombinants in ES cell clones targeted with two different constructs: one introduces a single nucleotide change in the PCNA gene and the other allows for a conditional inactivation of the wild-type PCNA allele. By using MLPA probes consisting of three oligonucleotides we were able to simultaneously detect and quantify both wild type and mutant alleles. PMID- 16340006 TI - Time-resolved fluorescence of 2-aminopurine as a probe of base flipping in M.HhaI DNA complexes. AB - DNA base flipping is an important mechanism in molecular enzymology, but its study is limited by the lack of an accessible and reliable diagnostic technique. A series of crystalline complexes of a DNA methyltransferase, M.HhaI, and its cognate DNA, in which a fluorescent nucleobase analogue, 2-aminopurine (AP), occupies defined positions with respect the target flipped base, have been prepared and their structures determined at higher than 2 A resolution. From time resolved fluorescence measurements of these single crystals, we have established that the fluorescence decay function of AP shows a pronounced, characteristic response to base flipping: the loss of the very short (approximately 100 ps) decay component and the large increase in the amplitude of the long (approximately 10 ns) component. When AP is positioned at sites other than the target site, this response is not seen. Most significantly, we have shown that the same clear response is apparent when M.HhaI complexes with DNA in solution, giving an unambiguous signal of base flipping. Analysis of the AP fluorescence decay function reveals conformational heterogeneity in the DNA-enzyme complexes that cannot be discerned from the present X-ray structures. PMID- 16340007 TI - The leucine rich region of DNA-PKcs contributes to its innate DNA affinity. AB - DNA-PK is a protein complex that consists of a DNA-binding, regulatory subunit [Ku] and a larger approximately 465 kDa catalytic subunit [DNA-PKcs], a serine/threonine protein kinase. The kinase activity of DNA-PKcs resides between residues 3745 and 4013, a PI3 kinase domain. Another recognized domain within this large protein is a leucine zipper (LZ) motif or perhaps more appropriately designated a leucine rich region (LRR) that spans residues 1503-1602. Whereas, DNA-PK's kinase activity has been shown to be absolutely indispensable for its function in non-homologous end joining (NHEJ), little is known about the functional relevance of the LRR. Here we show that DNA-PKcs with point mutations in the LRR can only partially reverse the radiosensitive phenotype and V(D)J recombination deficits of DNA-PKcs deficient cells. Disruption of the LRR motif affects the ability to purify DNA-PKcs via its binding to DNA-cellulose, but does not affect its interaction with Ku or its catalytic activity. These data suggest that the LRR region of DNA-PKcs may contribute to its intrinsic DNA affinity, and moreover, that intrinsic DNA binding is important for optimal function of DNA PKcs in repairing double strand breaks in living cells. PMID- 16340008 TI - Coupling DNA-binding and ATP hydrolysis in Escherichia coli RecQ: role of a highly conserved aromatic-rich sequence. AB - RecQ enzymes are broadly conserved Superfamily-2 (SF-2) DNA helicases that play critical roles in DNA metabolism. RecQ proteins use the energy of ATP hydrolysis to drive DNA unwinding; however, the mechanisms by which RecQ links ATPase activity to DNA-binding/unwinding are unknown. In many Superfamily-1 (SF-1) DNA helicases, helicase sequence motif III links these activities by binding both single-stranded (ss) DNA and ATP. However, the ssDNA-binding aromatic-rich element in motif III present in these enzymes is missing from SF-2 helicases, raising the question of how these enzymes link ATP hydrolysis to DNA binding/unwinding. We show that Escherichia coli RecQ contains a conserved aromatic-rich loop in its helicase domain between motifs II and III. Although placement of the RecQ aromatic-rich loop is topologically distinct relative to the SF-1 enzymes, both loops map to similar tertiary structural positions. We examined the functions of the E.coli RecQ aromatic-rich loop using RecQ variants with single amino acid substitutions within the segment. Our results indicate that the aromatic-rich loop in RecQ is critical for coupling ATPase and DNA binding/unwinding activities. Our studies also suggest that RecQ's aromatic-rich loop might couple ATP hydrolysis to DNA-binding in a mechanistically distinct manner from SF-1 helicases. PMID- 16340009 TI - Role of the N- and C-terminal extensions on the activity of mammalian mitochondrial translational initiation factor 3. AB - Mammalian mitochondrial translational initiation factor 3 (IF3(mt)) promotes initiation complex formation on mitochondrial 55S ribosomes in the presence of IF2(mt), fMet-tRNA and poly(A,U,G). The mature form of IF3(mt) is predicted to be 247 residues. Alignment of IF3(mt) with bacterial IF3 indicates that it has a central region with 20-30% identity to the bacterial factors. Both the N- and C termini of IF3(mt) have extensions of approximately 30 residues compared with bacterial IF3. To examine the role of the extensions on IF3(mt), deletion constructs were prepared in which the N-terminal extension, the C-terminal extension or both extensions were deleted. These truncated derivatives were slightly more active in promoting initiation complex formation than the mature form of IF3(mt). Mitochondrial 28S subunits have the ability to bind fMet-tRNA in the absence of mRNA. IF3(mt) promotes the dissociation of the fMet-tRNA bound in the absence of mRNA. This activity of IF3(mt) requires the C-terminal extension of this factor. Mitochondrial 28S subunits also bind mRNA independently of fMet tRNA or added initiation factors. IF3(mt) has no effect on the formation of these complexes and cannot dissociate them once formed. These observations have lead to a new model for the function of IF3(mt) in mitochondrial translational initiation. PMID- 16340010 TI - Priority setting in the REACH system. AB - Due to the large number of chemicals for which toxicological and ecotoxicological information is lacking, priority setting for data acquisition is a major concern in chemicals regulation. In the current European system, two administrative priority-setting criteria are used, namely novelty (i.e., time of market introduction) and production volume. In the proposed Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) system, the novelty criterion is no longer used, and production volume will be the main priority-setting criterion for testing requirements, supplemented in some cases with hazard indications obtained from QSAR modelling. This system for priority setting has severe weaknesses. In this paper we propose that a multicriteria system should be developed that includes at least three additional criteria: chemical properties, results from initial testing in a tiered system, and voluntary testing for which efficient incentives can be created. Toxicological and decision-theoretical research is needed to design testing systems with validated priority-setting mechanisms. PMID- 16340011 TI - Insulin antagonizes ischemia-induced Thr172 phosphorylation of AMP-activated protein kinase alpha-subunits in heart via hierarchical phosphorylation of Ser485/491. AB - Previous studies showed that insulin antagonizes AMP-activated protein kinase activation by ischemia and that protein kinase B might be implicated. Here we investigated whether the direct phosphorylation of AMP-activated protein kinase by protein kinase B might participate in this effect. Protein kinase B phosphorylated recombinant bacterially expressed AMP-activated protein kinase heterotrimers at Ser(485) of the alpha1-subunits. In perfused rat hearts, phosphorylation of the alpha1/alpha2 AMP-activated protein kinase subunits on Ser(485)/Ser(491) was increased by insulin and insulin pretreatment decreased the phosphorylation of the alpha-subunits at Thr(172) in a subsequent ischemic episode. It is proposed that the effect of insulin to antagonize AMP-activated protein kinase activation involves a hierarchical mechanism whereby Ser(485)/Ser(491) phosphorylation by protein kinase B reduces subsequent phosphorylation of Thr(172) by LKB1 and the resulting activation of AMP-activated protein kinase. PMID- 16340014 TI - The Niemann-Pick C1 protein in recycling endosomes of presynaptic nerve terminals. AB - Niemann-Pick type C (NPC) disease is a fatal, neurodegenerative disorder caused in 95% of cases by loss of function of NPC1, a ubiquitous endosomal transmembrane protein. A biochemical hallmark of NPC deficiency is cholesterol accumulation in the endocytic pathway. Although cholesterol trafficking defects are observed in all cell types, neurons are the most vulnerable to NPC1 deficiency, suggesting a specialized function for NPC1 in neurons. We investigated the subcellular localization of NPC1 in neurons to gain insight into the mechanism of action of NPC1 in neuronal metabolism. We show that NPC1 is abundant in axons of sympathetic neurons and is present in recycling endosomes in presynaptic nerve terminals. NPC1 deficiency causes morphological and biochemical changes in the presynaptic nerve terminal. Synaptic vesicles from Npc1(-/-) mice have normal cholesterol content but altered protein composition. We propose that NPC1 plays a previously unrecognized role in the presynaptic nerve terminal and that NPC1 deficiency at this site might contribute to the progressive neurological impairment in NPC disease. PMID- 16340015 TI - Crystal structure of a metal ion-bound IS200 transposase. AB - IS200 transposases, present in many bacteria and Archaea, appear to be distinct from other groups of transposases. To provide a structural basis for understanding the action of IS200 transposases, we have determined the crystal structure of the SSO1474 protein from Sulfolobus solfataricus, a member of the IS200 family, in both Mn(2+)-bound and Mn(2+)-free forms. Its monomer fold is distinct from other classes of structurally characterized transposases. Two monomers form a tight dimer by exchanging the C-terminal alpha-helix and by merging the two central beta-sheets into a large beta-sheet. Glu(55), His(62), and four water molecules provide the direct coordination sphere of the catalytically essential metal ion in the Mn(2+)-bound structure. His(16), Asp(59), and His(60) also play important roles in maintaining the metal binding site. The catalytic site is formed at the interface between monomers. The candidate nucleophile in the transposition mechanism, strictly conserved Tyr(121) coming from the other monomer, is turned away from the active site, suggesting that a conformational change is likely to occur during the catalytic cycle. PMID- 16340016 TI - Phosphate-binding tag, a new tool to visualize phosphorylated proteins. AB - We introduce two methods for the visualization of phosphorylated proteins using alkoxide-bridged dinuclear metal (i.e. Zn(2+) or Mn(2+)) complexes as novel phosphate-binding tag (Phos-tag) molecules. Both Zn(2+)- and Mn(2+)-Phos-tag molecules preferentially capture phosphomonoester dianions bound to Ser, Thr, and Tyr residues. One method is based on an ECL system using biotin-pendant Zn(2+) Phos-tag and horseradish peroxidase-conjugated streptavidin. We demonstrate the electroblotting analyses of protein phosphorylation status by the phosphate selective ECL signals. Another method is based on the mobility shift of phosphorylated proteins in SDS-PAGE with polyacrylamide-bound Mn(2+)-Phos-tag. Phosphorylated proteins in the gel are visualized as slower migration bands compared with corresponding dephosphorylated proteins. We demonstrate the kinase and phosphatase assays by phosphate affinity electrophoresis (Mn(2+)-Phos-tag SDS PAGE). PMID- 16340017 TI - Underreported, underacknowledged: child abuse can no longer be ignored. PMID- 16340018 TI - A community-based study of the prevalence of constipation in young children and the role of dietary fibre. AB - OBJECTIVES: To study the prevalence of constipation in young children, and to explore its association with dietary fibre intake. DESIGN: Cross-sectional community-based study. SETTING: Three kindergartens randomly selected from the eastern district of Hong Kong. PARTICIPANTS: Between October 2003 and January 2004, parents of children aged 3 to 5 years completed a specially designed questionnaire and a 3-day dietary record form, which were used to collect information on bowel function and dietary intake. Children with constipation were identified based on Rome criteria. Children with normal bowel habits served as a comparison group. MAIN OUTCOME MEASURES: Dietary intake of energy, protein, and dietary fibre. RESULTS: Of 778 children recruited, 561 complete sets of data were successfully obtained from the participating kindergartens. One hundred and sixty six (29.6%) children were found to be constipated. The incidence of a family history of constipation was significantly higher in the constipated group (14%) than in the non-constipated group (7%) [P=0.013]. Mean dietary fibre consumption was 4.1 g/d (standard deviation, 2.3 g/d) in all children corresponding to 45.5% (standard deviation, 24.9%) of the daily recommendation. Constipated children (mean, 40.7%; standard deviation, 20.5%) had a significantly lower dietary fibre intake of the daily recommendation than the non-constipated group (mean, 47.5%; standard deviation, 26.2%) [P=0.017]. CONCLUSION: Up to 30% of preschool children in the eastern district had constipation. A family history of constipation was related to its occurrence in the studied children. Dietary fibre intake was insufficient in all children and even lower in those who were constipated. PMID- 16340019 TI - Epidemiological study of diabetic retinopathy in a primary care setting in Hong Kong. AB - OBJECTIVES: To estimate the prevalence and risk factors of diabetic retinopathy in type 2 diabetic patients, and to investigate the difference in retinopathy progression in patients with normal fundi or established retinopathy at baseline and the risk factors implicated in the progression. DESIGN: Retrospective community-based study. SETTING: Ten primary care clinics in Hong Kong. PATIENTS: Type 2 diabetic patients; subsidiary analysis included subjects with more than one screening event. MAIN OUTCOME MEASURES: Patient demographics, baseline prevalence, and risk factors of diabetic retinopathy; progression of retinopathy in patients with normal fundi and established retinopathy at baseline, and the associated risk factors. RESULTS: A total of 6165 patients were recruited from January 1998 to May 2004. Primary analysis included 4423 patients with good quality retinal photographs. The mean age of the patients was 60.36 years (standard deviation, 10.80 years; range, 28-94 years), the mean duration of diabetes was 4.71 years (standard deviation, 4.67 years; range, 0.1-40.6 years), and the mean level of glycated haemoglobin was 7.47% (standard deviation, 1.44%). The prevalence of retinopathy at baseline was 28.4%. Subsidiary analysis showed progression to sight-threatening retinopathy was more common in the group with baseline retinopathy than that without (7.9% vs 0.7%), and occurred at a faster rate (mean, 1.5 [range, 0.5-3.0] vs 2.0 [1.0-4.2] years). Logistic regression revealed that the level of glycated haemoglobin was positively associated with both the onset (P<0.001) and progression of retinopathy (P=0.03). CONCLUSION: Optimal glycaemic control is important for reducing sight-threatening retinopathy. Close observation is required for patients with established retinopathy as progression occurs more rapidly. PMID- 16340020 TI - Fractures associated with non-accidental injury--an orthopaedic perspective in a local regional hospital. AB - OBJECTIVES: To identify the potential risk factors for fractures due to non accidental injury in children, and to alert clinicians and health care workers to the diagnosis of child abuse. DESIGN: Retrospective review. SETTING: Regional hospital, Hong Kong. PATIENTS: Children who were admitted to the hospital with a clinical diagnosis of child abuse with associated fracture(s) between January 1996 and April 2004. MAIN OUTCOME MEASURES: Demographic data, site of fractures, and investigations performed. RESULTS: Of the 377 children presented with non accidental injuries, 29 (15 male, 14 female) had bone fractures. The mean age of the 29 children at the time of injury was 5 years and 5 months. Of the nine records showing pregnancy, seven were unplanned. Approximately 75% of the families were living in public housing estates, and 28% were receiving social security subsidy. Over half (52%) of the abused children were aged 3 years or less. A total of 78 fractures were documented with a mean of 2.7 fractures per child. The most common sites of fracture were the forearm (29%), followed by the ribs (24%). Most long bone fractures occurred in those aged 3 years or less. Fractures were detected by skeletal survey and bone scan for 90% of the children. CONCLUSION: A number of potential risk factors were identified in children with fractures associated with non-accidental injury. They included age younger than 3 years, lower socio-economic status, presentation with long bone fracture, and unplanned pregnancy. Bone scan and skeletal survey are mutually complementary, and both should be performed in cases of suspected child abuse. Subsequent management requires cooperation of multi-disciplinary health care professionals. PMID- 16340021 TI - Temozolomide in the treatment of recurrent malignant glioma in Chinese patients. AB - OBJECTIVE: To determine the anti-tumour efficacy and safety profile of temozolomide in local Chinese patients with recurrent malignant glioma. DESIGN. Open-label trial. SETTING: University teaching hospital, Hong Kong. PATIENTS: Twenty-two patients had been enrolled in the study since 2001. Patients had to show unequivocal evidence of tumour recurrence or progression on gadolinium enhanced magnetic resonance imaging after failing conventional radiotherapy and surgery for initial disease. Histology reviewed by a neuropathologist was required to show anaplastic glioma (anaplastic astrocytoma, anaplastic oligodendroglioma, or mixed anaplastic oligoastrocytoma) or glioblastoma multiforme. INTERVENTIONS: Patients were treated with temozolomide (200 mg/m(2) per day for the first 5 days of a 28-day cycle for four cycles) and monitored clinically every month and radiologically (gadolinium magnetic resonance imaging) at 6 months. MAIN OUTCOME MEASURES: Six-month progression-free survival and objective response rate. RESULTS: Twenty-two patients with recurrent malignant glioma were recruited between January 2001 and July 2004. Progression-free survival at 6 months was 54.5%. The mean progression-free survival for all patients was 7.2 months. The objective response rate, determined by gadolinium magnetic resonance imaging, was 9% for patients demonstrating a complete or partial response and a further 45% for patients demonstrating stable disease. Temozolomide was well tolerated orally with minimal adverse events. CONCLUSION. Preliminary results showed that temozolomide had an acceptable safety profile and anti-tumour activity in recurrent malignant glioma in local Chinese population. The results were comparable with those of western studies. PMID- 16340022 TI - Cerebrospinal fluid to serum glucose ratio in non-hypoglycorrhachic neurological conditions. AB - OBJECTIVE: To explore the relevance of cerebrospinal fluid to serum glucose ratio in non-hypoglycorrhachic conditions. DESIGN: Retrospective observational study. SETTING: Neurology ward, university teaching hospital, Hong Kong. PATIENTS: Adult patients with conditions unrelated to hypoglycorrhachia who underwent lumbar puncture. MAIN OUTCOME MEASURES: Cerebrospinal fluid and simultaneous serum glucose concentrations, and their ratio to each other. RESULTS: Between September 1998 and August 2003, 170 cerebrospinal fluid and serum glucose samples were collected from 138 patients. Mean cerebrospinal fluid to serum glucose ratio was 0.61 (standard deviation, 0.142; range, 0.21-1.00). With the exception of cerebrospinal fluid protein level, laboratory parameters were similar among different diseases. The glucose ratio was lower than 0.6 in 43% and lower than 0.5 in 19% of samples. Cases with a low glucose ratio appeared to have higher serum glucose concentrations (significant among groups with different glucose ratios, P<0.001). The mean glucose ratio (0.65) was also significantly higher in patients with serum glucose concentration of lower than 7.8 mmol/L compared with those with serum glucose concentration between 7.8 and 11.1 mmol/L (mean, 0.46), or higher than 11.1 mmol/L (mean, 0.46) [P<0.001]. There was a strong negative correlation between the glucose ratio and serum glucose concentration (r= -0.704, P<0.001). CONCLUSION: A lowered cerebrospinal fluid to serum glucose ratio is often seen in the absence of an appropriate disorder, especially when simultaneous serum glucose concentration is elevated. This may be explained by the saturation kinetics of glucose transportation in hyperglycaemia, and the time lag for cerebrospinal fluid and glucose to equilibrate when the blood level fluctuates. PMID- 16340023 TI - An active surveillance study of vancomycin-resistant Enterococcus in Queen Elizabeth Hospital, Hong Kong. AB - OBJECTIVE: To assess the rate of faecal vancomycin-resistant Enterococcus colonisation in high-risk patients in a regional hospital. DESIGN: Prospective observational surveillance study. SETTING: Queen Elizabeth Hospital, Hong Kong. PATIENTS: From September 2001 to December 2002, stool samples from patients in the intensive care unit and patients in whom Clostridium difficile testing was requested were used for study using a broth enrichment method. MAIN OUTCOME MEASURES: Number of faecal vancomycin-resistant Enterococcus colonisation. RESULTS: A total of 2414 cultures from 1792 patients were tested for vancomycin resistant Enterococcus using a broth enrichment method. Only one (0.06%) patient was found to harbour a vancomycin-resistant Enterococcus faecalis in the gastro intestinal tract. Surveillance cultures from contacts of the case revealed another six with vancomycin-resistant Enterococcus faecalis. Vancomycin-resistant Enterococcus faecalis was also later reported from a clinical specimen (catheterized urine) of another patient. They were all epidemiologically linked to the index case. Mean inhibitory concentrations of vancomycin and teicoplanin were determined to be higher than 256 and 0.5 microgram/mL, respectively by E test for all the vancomycin-resistant Enterococcus isolates. Polymerase chain reaction analysis confirmed the presence of vanB genes and the result was in line with the phenotype. Pulsed-field gel electrophoresis confirmed a monoclonal vancomycin-resistant Enterococcus outbreak. Strict infection control measures recommended by the Centers for Disease Control and Prevention were followed and the outbreak was successfully controlled. CONCLUSION: Vancomycin-resistant Enterococcus colonisation is rare, but present among high-risk patients in our hospital. A routine surveillance programme should be implemented that will enable early case detection and prompt initiation of infection control measures to prevent the emergence of an endemic situation. PMID- 16340024 TI - Barriers to participation in a phase II cardiac rehabilitation programme. AB - OBJECTIVES: To identify barriers to participation in a phase II cardiac rehabilitation programme and measures that may enhance participation. DESIGN: Prospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Cardiac patients recruited for a phase I cardiac rehabilitation programme from July 2002 to January 2003. MAIN OUTCOME MEASURES: Reasons for not participating in a phase II cardiac rehabilitation programme. RESULTS: Of the 193 patients recruited for a phase I cardiac rehabilitation programme, 152 (79%) patients, with a mean age of 70.3 years (standard deviation, 11.9 years), did not proceed to phase II programme. Eleven (7%) deaths occurred before commencement of phase II and 74 (49%) patients were considered physically unfit. Reasons for the latter included fractures, pain, or degenerative changes in the lower limbs (24%), and co morbidities such as cerebrovascular accident (19%), chronic renal failure (11%), congestive heart failure (9%), and unstable angina (8%). Phase II rehabilitation was postponed until after completion of scheduled cardiac interventions in 13% of patients. Failure of physicians to arrange the pre-phase II exercise stress test as per protocol was reported in 7% of patients. Other reasons were reported: work or time conflicts (16%), non-compliance with cardiac treatment (5%), financial constraints (4%), self-exercise (3%), fear after exercise stress testing (3%), and patients returning to their original cardiologists for treatment (3%). CONCLUSIONS: A significant (79%) proportion of patients did not proceed to a phase II cardiac rehabilitation programme for a variety of reasons. These included physical unfitness, work or time conflicts, and need to attend scheduled cardiac interventions. Further studies are required to determine how to overcome obstacles to cardiac rehabilitation. PMID- 16340025 TI - Parkinson's disease: aetiology, diagnosis, and management. AB - OBJECTIVE: To review the aetiology, diagnosis, and management of Parkinson's disease, with a local perspective. DATA SOURCES: Medline from 1966 onwards, and all major neurological journals and movement disorder journals were searched for evidence on the aetiology, diagnosis, and management of Parkinson's disease. STUDY SELECTION: Key words for the literature search were "Parkinson's disease" and "Chinese" or "Hong Kong". DATA EXTRACTION: All relevant articles in English were reviewed. DATA SYNTHESIS: The number of promising genes for familial Parkinson's disease is still expanding rapidly and there has been a wealth of studies on susceptibility genes for Parkinson's disease. Potential treatment choices include the use of agents thought to be neuroprotective, symptomatic treatment with drugs or surgery, and non-pharmacological treatments. Pharmacological treatment using a dopa-sparing strategy and continuous dopaminergic stimulation is now gaining support to address the issue of long-term motor complications. Surgical treatment with deep brain stimulation is safe and effective for refractory cases and has been increasingly utilised locally. CONCLUSIONS: Medical therapy remains the mainstay of treatment and newer agents and treatment approaches are emerging, which will hopefully address the issue of neuroprotection and provide symptomatic treatment with fewer motor complications. PMID- 16340026 TI - Bone loss associated with long-term use of depot medroxyprogesterone acetate. AB - On 17 November 2004, the United States Food and Drug Administration issued a black box warning on the long-term use of depot medroxyprogesterone acetate stating that bone loss might be irreversible with use of more than 2 years. Despite the seriousness of such a safety warning, the Food and Drug Administration provided no clinical recommendations. Various professional bodies have made different recommendations on the management of women prescribing such long-term injections but there is no consensus on the best practice. Thus individual institutions need to revise service protocols. The Health Services Subcommittee of the Family Planning Association of Hong Kong has reviewed the scientific evidence from international and local studies and made its recommendations in a meeting held on 1 March 2005. This article aimed to share our opinion with other medical professionals in Hong Kong. PMID- 16340027 TI - Treatment of epiretinal membrane: an update. AB - Epiretinal membrane is a common surgical disease of the elderly. The aetiology, clinical presentation, and diagnosis are briefly described. Management, principally vitrectomy and membrane removal, is then elaborated. Recent advances such as internal limiting membrane peeling are further discussed. The safety and efficacy of intra-ocular dyes, including indocyanine green and trypan blue, as well as the new technique of 'double staining', are evaluated. PMID- 16340028 TI - Recurrent lower limb embolism from thoracic aortic mural thrombus: a rare presentation of occult malignancy. AB - Initial presentation of a malignant disease as recurrent attacks of lower limb ischaemia due to emboli from a mural thrombus in the descending thoracic aorta is extremely rare. A diagnosis of malignancy may thus easily be overlooked. Recent advances in imaging technology have made the diagnosis of thoracic aortic mural thrombi much easier. Occult malignancy should always be suspected in the absence of biochemical evidence of hypercoagulability. We report on a patient with underlying malignant disease who presented with lower limb ischaemia that was relieved by axillobifemoral bypass. PMID- 16340029 TI - A case of tetramine poisoning: a lethal rodenticide. AB - We report a fatal case of suicide presenting with convulsions and subsequently multi-organ failure. Rodenticide poisoning was not suspected until the next day when tetramine was detected in the patient's blood, urine, and food residues. Tetramine is a potent rodenticide with a rapid action and high mortality. The poison has been banned worldwide but is still readily available in Mainland China. Outbreaks of poisoning are reported frequently and doctors should be alert for this lethal toxin. PMID- 16340030 TI - Tetramine poisoning. AB - We report on two patients who presented with status epilepticus due to ingestion of rat poison containing tetramine. Both had eaten the same meal, subsequently presumed to be the source of the poison. Physical examination and investigation were unremarkable and diagnosis was based on patient history. Seizures were ultimately controlled with ketamine, after unsuccessful attempt of benzodiazepine and sodium thiopentone. One week after poisoning, both patients underwent one session of high-volume haemofiltration followed by charcoal haemoperfusion to eliminate the toxin from the body. Plasma tetramine levels then decreased from 0.95 microgram/mL to 0.35 microgram/mL and from 0.53 microgram/mL to 0.40 microgram/mL, respectively. PMID- 16340031 TI - Disseminated cutaneous infection with Mycobacterium chelonae mimicking panniculitis in a patient with dermatomyositis. AB - We report a case of disseminated cutaneous Mycobacterium chelonae infection. A patient with dermatomyositis associated with malignancy presented with features of panniculitis. This was later confirmed to be cutaneous Mycobacterium chelonae infection. Disseminated cutaneous Mycobacterium chelonae infection and panniculitis are reviewed. PMID- 16340032 TI - Acute anticholinergic poisoning in children. AB - We report two cases of unintentional poisoning with anticholinergic agents. The first patient, a 7-year-old girl, was prescribed four different medications by a general practitioner for treatment of abdominal colic and diarrhoea. All drugs had anticholinergic properties. The second patient, a 16-month-old boy, ingested his mother's cyproheptadine tablets. Both children presented with central and peripheral symptoms and signs compatible with acute anticholinergic syndrome. They recovered spontaneously following intravenous fluid replacement and close observation. Gastric lavage was also performed on the boy. Poisoning with cholinergic antagonists in children is a potentially serious hazard in Hong Kong. It may be avoided by careful prescribing on the part of general practitioners and safe storage of all medicinal products in the home environment. PMID- 16340034 TI - Doctors as fiduciaries: a legal construct of the patient-physician relationship. PMID- 16340033 TI - Magnetic resonance venogram of intravenous leiomyomatosis. PMID- 16340035 TI - Neuro-hormonal control of food intake: basic mechanisms and clinical implications. AB - Obesity is one of the most common metabolic diseases and the greatest threats of the health because of possibility of numerous complications. In order to design effective drugs or apply the helpful surgical procedure it is essential to understand physiology of appetite control and pathophysiology of obesity. According to the first law of thermodynamics, the energy input in the form of food, equals energy expenditure through exercise, basal metabolism, thermogenesis and fat biosynthesis. The control of body weight actually concerns the control of adipose tissue with the key role of hypothalamus, possessing several neuronal centers such as that in lateral hypothalamic nuclei considered to be "hunger" center and in ventromedial nuclei serving as the "satiety" center. In addition, paraventricular and arcuate hypothalamic nuclei (ARC) are the sites where multiple hormones, released from the gut and adipose tissue, converge to regulate food intake and energy expenditure. There are two distinct types of neurons in ARC that are important in control of food intake; (1) preopiomelanocortin (POMC) neurons activated by an orexigenic hormones and releasing alpha-melanocyte stimulating hormone (alpha-MSH) in satiety center and (2) neurons activated by orexigenic peptides such as ghrelin that release the substances including neuropeptide Y (NPY) and Agouti-Related Peptide (AgRP) in hunger center. ARC integrates neural (mostly vagal) and humoral inputs such as enteropeptides including orexigenic (ghrelin and orexins) and an orexigenic peptides (cholecystokinin, polypeptide YY, glucagon-like peptide-1, oxyntomodulin, leptin and others) that exert a physiological role in regulating appetite and satiety. The peripherally (gut, adipose tissue) and centrally expressed modulators of appetitive behavior act through specific receptors in the afferent (mostly vagal) nerves and hypothalamic neurons implicated in adiposity signaling and regulation of food intake. PMID- 16340036 TI - Influence of vagal nerve stimulation on food intake and body weight--results of experimental studies. AB - The paper reviews recent advances in vagal nerve stimulation for the control of food intake and body weight. The vagal nerves are the predominant pathway in the "brain-gut axis" responsible for short term regulation of food intake. Stimulation of afferent vagal traffic attenuates food intake by vagal projections to nucleus tractus solitarius, arcuate nucleus and its convergence's to thalamic center of satiety. A few studies have been published in this field so far. All of them are consistent and show significant decrease in body mass during vagal stimulation. Due to promising results of experimental studies, clinical trials are expected in the near future. PMID- 16340037 TI - The progress in bariatric surgery. AB - Morbid obesity, caused by fat tissue accumulation, is a serial multi-factorial chronic disease, with rapidly increasing prevalence in most countries in the world including Poland. Conservative treatment of morbid obesity is almost always unsatisfactory and that is why several surgical methods have been developed. There are four kind of methods: malabsorbtive procedures; restrictive procedures; malabsorbtive/restrictive procedures and experimental procedures. The development of bariatric surgery goes back to 1952 and since that time it has been evolving dynamically. All the surgical methods have benefits and disadvantages. Presently the introduction of minimally invasive surgical techniques seems to be very safe, efficient and cost-effective in treatment for morbid obesity. New methods are also being evaluated, such as gastric myo-electrical stimulation. Bariatric surgery will still be developing until we understand all the factors responsible for it is origin. PMID- 16340038 TI - Metabolic and antioxidative changes in liver steatosis induced by high-fat, low carbohydrate diet in rabbits. AB - Significantly decreased amounts of carbohydrates and increase of fat in diet (Atkins' diet, Jan Kwasniewski's diet) may interfere with liver metabolism because main source of liver energy are carbohydrates. Large amount of fatty acid in diet may induce overproduction of reactive oxygen species in mitochondria because of increased oxidation and may cause steatosis. The influence of high fat, low-carbohydrate diet (contained 14 g% protein, 25 g% fat, 8 g% carbohydrate) and compared to usual habitual diet (13 g% protein, 3 g% fat, 28 g% carbohydrate) as a control on liver function was examined in adult male rabbits. Fodder and water was ad libitum. The experiment lasted 24 weeks. At the beginning and every six weeks rabbits were weighed and blood was taken. Plasma glucose and cholesterol concentration, activity of glutamate dehydrogenase, alanine and aspartate in plasma and liver did not change. Concentrations of triacylglycerols (TG) in plasma were lower in the study group. Activity of aldolase increased in plasma and in liver in the study group while activity of sorbite dehydrogenase decreased in plasma at the end of the experiment. Malondialdehyde (MDA) concentration increased in aorta with no changes in liver and erythrocytes. Activity of glutathione peroxidase increased in erythrocytes and liver while total SOD increased only in the liver. High-fat, low-carbohydrate diet despite the lack of growth of the body mass, modifies significantly the homeostasis of carbohydrates and antioxidants in liver and enhanced production of TG in this organ, resulting in its steatosis. PMID- 16340039 TI - Sleep study in patients with overweight and obesity. AB - Obstructive sleep apnea syndrome (OSAS) is a disorder characterized by repetitive collapse of the pharyngeal airway during sleep, which leads to oxygen desaturation, sleep fragmentation and daytime somnolence. Obesity is one of the most important risk factor for the development of OSAS. The exact mechanisms responsible for the relationship between obesity and OSAS are still unclear. The fat deposits in the pharynx region as well as the reduction in the lung volume have been considered as factors that might be responsible for the increase of the upper airway collapsibility. The aim of our study was to evaluate the correlation between the Body Mass Index (BMI) and sleep study parameters in overweight and obese patients suffering from breathing disturbances during sleep. We studied a group of 106 consecutive obese or overweight patients with a primary complaint of snoring or other breathing disturbances during sleep. In all cases, BMI and sleep studies (PolyMESAM) were examined. We evaluated relationship between the BMI and sleep study parameters such as Respiratory Disturbance Index (RDI), Apnea Index (AI), Desaturation Index (DI) and Average of Lowest Saturation (LSAT). The results showed the lack of significant statistical correlations between BMI and all the sleep parameters studied in the overweight patients and the statistical positive correlation between the BMI and RDI in the obese cases. We conclude that BMI determination may be considered as a simple, yet important predictor, of the OSAS in the group of obese patients. PMID- 16340040 TI - The nasal airway evaluation in morbid obesity. AB - The aim of the present study was to evaluate objectively the nasal patency in the obese patients. A total of 18 morbidly obese patients were recruited for the study. All of them were surgically treated because of morbid obesity using Bilo Pancreatic Diversion (BPD) or Laparoscopic Gastric Banding (LGB) methods. The patients were free of nasal abnormalities, such as septum deviation, polyps, nasal concha hypertrophy and paranasal sinus diseases. This group comprised 10 men and 8 women aged from 17 to 54. The mean Body Mass Index (BMI) was 51.6 kg/m(2), ranged from 34.7 to 61.8 kg/m(2). In all of the patients the nasal patency was examined by active anterior rhinomanometry according International Standardization Rhinomanometric Committee using air pressure 75, 100 and 150 dPa. The results were compared to the healthy control group. The correlation between BMI and nasal airflow pressure was also examined. We found that inspiration values for 75, 100 and 150 dPa as well as the expiration values for 100 and 150 dPa in standard method and expiration values for 100 and 150 dPa in Broms method using anterior rhinomanometry in morbidly obese patients were statistically significant higher in comparison with the healthy controls. No statistical significant correlation between BMI of obese patients and the airflow pressure values was found. We conclude that in the morbid obesity the nasal patency is reduced as compared to the healthy controls. PMID- 16340041 TI - Helicobacter pylori in the oral cavity and its implications for gastric infection, periodontal health, immunology and dyspepsia. AB - Helicobacter pylori (H. pylori) is an important gastrointestinal pathogen associated with gastritis as well as gastric or duodenal ulcers and gastric cancer. The oral cavity has been considered as a potential reservoir for the gastric infection and reinfection. The objective of our studies was to evaluate the influence of oral H. pylori for the stomach infection and the release of gut hormones affecting food intake such as ghrelin and gastric secretion such as gastrin. Additionally, the contribution of H. pylori in the periodontal disease has been examined. H. pylori infection in stomach was assessed by (13)C- Urease Breath Test and presence of the bacteria in oral cavity by culture. The periodontal status was measured by pockets depth with the periodontal probe. We estimated the serum level of IgG anti-H. pylori, anti-VacA, anti-CagA, ghrelin, gastrin, TNF-alpha and IL-8 in blood and the level of IgA anti-H. pylori in saliva. The presence of H. pylori in oral cavity was detected in 54.1% of examined individuals, whereas the H. pylori gastric infection in tested group was found in 51% cases. However, the correlation analysis between those two groups of patients involving together about 100 subjects showed that within the group of patients with positive gastric H. pylori infection only 45.1% did not show the presence of H. pylori in saliva and 43.1% showed no H. pylori in supragingival plaque. In line of these findings patients who did not have gastric H. pylori infection, 53.2% showed presence of H. pylori in saliva and 42.9% in supragingival plaques. Serum level of ghrelin and gastrin in subjects with oral H. pylori inoculation but without gastric H. pylori infection were not significantly different from those without the presence of this germ in oral cavity. In contrast, gastric H. pylori infection resulted in significant reduction in serum ghrelin levels and significant elevation of gastrin as compared to those who were gastric H. pylori negative. We concluded that oral H. pylori alone does not seem to serve as bacterium sanctuary for gastric H. pylori infection and, unlike gastric infection, it fails to affect serum levels of hormones stimulating appetitive behaviour such as ghrelin and gastric acid secretion such as gastrin. PMID- 16340043 TI - Effect of extremely low frequency of electromagnetic fields on cell proliferation, antioxidative enzyme activities and lipid peroxidation in 3T3-L1 preadipocytes--an in vitro study. AB - The exposure to extremely low frequency electromagnetic field (ELF-MF, frequencies less than 200-300 Hz) can alter the transcription and translation of genes, influence the cell proliferation rate and affect enzyme activities. Moreover, the hypothesis that ELF-MF increases free oxygen metabolites generation has been proposed. Since recent in vivo studies suggest that electric and magnetic fields are able to affect adipose cells metabolism. The aim of the study was to examine the effects of ELF-MF (frequency of basic impulse 180-195 Hz, induction 120 microT) on cell proliferation, antioxidative enzyme activities and malondialdehyde (MDA) concentration in 3T3-L1 preadipocyte cell culture. We found that ELF-MF application lasting 36 minutes daily failed to influence cell count after 24h and 48 h of incubation. After 24 h, in the ELF-MF treated group, manganese- and copper-zinc-containing superoxide dismutase (MnSOD and Cu/ZnSOD) isoenzymes media activities were decreased, catalase activity was increased, whereas there were no significant differences in glutathione peroxidase (GSH-Px) and glutathione reductase (GSSG-Rd) activities in comparison to the control. After 48 h of incubation, all enzyme activities were reduced, except for GSSG-Rd, in which no changes were noticed. MDA concentration at 24 h after incubation with the exposure to ELF-MF was significantly higher in comparison to the control, without ELF-MF. After 48 h of incubation, MDA levels were significantly lower in both groups with no differences between the groups without and with ELF-MF. We conclude that ELF-MF influences antioxidative enzyme activities and increases lipid peroxidation in 3T3-L1 preadipocyte cultures. PMID- 16340042 TI - Influence of melatonin on cell proliferation, antioxidative enzyme activities and lipid peroxidation in 3T3-L1 preadipocytes--an in vitro study. AB - Melatonin, acting via MT1, MT2 and MT3 membrane receptors, influences central and peripheral regulatory mechanisms of energy homeostasis in mammals. In peripheral tissues, it evokes the pro-proliferative effect in a number of normal cells. Moreover, this hormone inhibits lipolysis in subcutaneous adipocytes in vitro and reduces free oxygen metabolites-induced damage acting directly, as a free radical scavenger, and indirectly, by stimulation of antioxidative enzyme activities. The aim of the study was to examine the effects of melatonin on cell proliferation, antioxidative enzyme activities and malondialdehyde (MDA) concentration in 3T3-L1 preadipocyte cell culture. We found that melatonin (10(-3) and 10(-6) M/L) stimulated cell proliferation in dose- and time-depending manner, and this effect was inhibited by a relatively selective MT2 receptor antagonist - luzindole (10( 4) M/L). Melatonin, increased activities of manganese containing and copper-zinc containing superoxide dismutase (MnSOD and Cu/ZnSOD) isoenzymes, catalase, glutathione reductase and glutathione peroxidase after 24 h of incubation. In contrast, after 48 h of incubation, activities of all studied enzymes were lower than in the control group. There were no changes in MDA concentrations after 24 h of incubation, whereas, in melatonin-treated media, after 48 h of the experiment, MDA level was significantly decreased. Our results demonstrate that melatonin, acting via MT2 receptors, stimulates proliferation of 3T3-L1 preadipocytes and this action could be due to the enhancement in antioxidative enzyme activities and attenuation of lipid peroxidation by this indole. PMID- 16340044 TI - Bariatric surgery in Poland from 1993 to 2003. AB - Morbid obesity affects about 300, 000 patients in Poland. The number and type of bariatric procedures performed between 1993 and 2003 has been based on data collected from Polish surgeons active in this field. During the years 1993 - 2003, 1285 primary bariatric procedures were performed in total. Amongst these 79.1% were, commonly done as restrictive ones, including vertical banded gastroplasty-76.3%, laparoscopic adjustable gastric banding-17.7% and silastic ring vertical gastroplasty-5.5% and at last non-adjustable gastric band with 0.5%. Less popular are malabsorptive procedures - 20.9%, with Roux-en-Y gastric bypass - 79.1% and biliopancreatic diversion 20.9%. During last year 2003: malabsorptive procedures represented one third of all primary procedures and the laparoscopic approach was utilised in 18 % of operations. 61 revisions were reported. There are only three centres with experience in treating more than 100 patients (one of them treated over 600). Two centres have experience in carrying out bariatric surgery for longer than five years. During 1993 - 1997 216 operations were performed. Between 1998 - 2002: 724 procedures, and during last year 345. Patients were treated during last year in ten surgical departments. The number of morbidly obese patients treated surgically in Poland is increasing, but it is still inadequate to meet the growing demand. PMID- 16340045 TI - Biliopancreatic diversion in Poland. AB - Biliopancreatic diversion (BPD) is a bariatric operation of proved efficacy especially in patients with metabolic comorbidities. The aim was to assess the results of BPD in relation to weight loss and coexisting lipid and glucose metabolism disturbances in the Polish population. Between October 2001 and December 2003 57 morbidly obese patients (24 male and 33 female) underwent BPD. Median age was 46 years (range: 17-58) and median body mass index (BMI) was 50.2 kg/m(2) (range: 40.1-73 kg/m(2)). Type 2 diabetes mellitus was observed in 11 patients (19.3%), hypercholesterolaemia in 38 patients (66.7%) and hypertriglyceridaemia in 37 patients (64.9%). Median value of BMI decreased at three months to 40.0 kg/m(2), at six months to 36.7 kg/m(2), and after a year to 32.3 kg/m2. Diabetes was completely resolved in all patients. After six months, triglycerides and cholesterol levels were higher than normal only in one patient. Early specific complications that manifested as seroma and nosocomial pneumonia occurred in 5.3% and 3.5% patients, respectively. The following specific late complications were observed: ulceration of stomach stump (3.4%), anaemia (14.0%), hypoalbuminaemia (8.8%), deterioration of haemorrhoids (15.8). All of above complications were treated conservatively expect two haemorrhoidectomies. One patient died due to myocardial infarction eight months after BPD. Five cases of incisional hernias (8.8%) were found. BPD is an effective bariatric procedure also in the Polish population, resulting not only in weight loss but also in the improvement of lipid and glucose metabolism. PMID- 16340046 TI - Health-related quality of life after the surgical treatment of obesity. AB - The aim of the study is to answer the question: what is the range and character of changes within health-related quality of life (HRQL) in obese patients who have undergone surgical treatment (VBG - vertical banded gastroplasty or RYGBP - Roux-en-Y gastric bypass) in the period of three and six months after operation? Three groups of patients participated in the study: G I (n=33; BMI 49.90 +/- 6.74 kg/m(2)) - tested prior to the operation, G II (n=17; BMI 34.26 +/- 5.28 kg/m(2)) - tested six months after the operation, and G III (n=11) - examined prior to the operation (BMI 45.72 +/- 5.27 kg/m(2)) as well as three months after the operation (BMI 36.10 +/- 4.60 kg/m(2)). The Nottingham Health Profile (NHP) questionnaire was used as an indicator of HRQL. Surgical treatment of obesity resulted in significant positive changes in relation to the basic dimensions of NHP: energy, pain, physical mobility and in the influence of the current health state on selected spheres of life: paid employment, jobs around the house, social life, sex life, interests and hobbies, holidays. Significant changes in HRQL resulting from bariatric surgery occur in a relatively short period of time and are related not only to physical but also to social functioning. PMID- 16340047 TI - Results of revisional operation following vertical banded gastroplasty performed due to morbid obesity--comparison between restoration of vertical banded gastroplasty and conversion of gastric bypass up to three years. AB - The aim of this study was to analyse incidence and efficacy of revisional surgery for failed vertical banded gastroplasty among 458 patients who underwent primary surgery between 1993 and 2003. Staple line disruption was diagnosed in 29 patients and was an indication for restoration of gastroplasty in 10 cases and a conversion to Roux-en-Y gastric bypass in 19 patients. In two cases of outlet stenosis the band was exchanged to enlarge the collar. In two cases of psychological intolerance of restriction the band was removed because of refusion by patients the conversion to Roux-en-Y gastric bypass. A substantial weight reduction without statistical differences between restoration and conversion group was recognized. In two patients (20%) after restoration and three patients (15.8%) after conversion we observed weight regain (p=0.57). In cases with removed band weight regained up to its value recorded before surgery. In patients with exchanged band weight was under control. No serious complications were observed. We could conclude that patients with weight regain after vertical banded gastroplasty should be offered conversion to Roux-en-Y gastric bypass. When malabsorption is refused, restoration of vertical banded gastroplasty could be also performed. Both of procedures are technically difficult but safe. PMID- 16340048 TI - Novel therapies need full outcome assessments. PMID- 16340049 TI - Evaluation of an Internet delivered pediatric diagnosis support system (ISABEL) in a tertiary care center in India. AB - BACKGROUND: Young graduates manning the emergency rooms in public hospitals often need guidance in diagnosing critically ill patients due to their limited clinical experience. Textbooks, manuals and several websites are of limited assistance, as they do not generate patient-specific advice. ISABEL diagnostic tool, an Internet delivered pediatric diagnosis support system that provides such information has not been evaluated in developing countries. AIM: To study the sensitivity of the ISABEL diagnostic tool. MATERIAL AND METHOD: Records of patients admitted in the pediatric intensive care unit in a metropolitan hospital in India during January 2000-July 2002 were retrieved. Resident medical officers wrote key clinical and laboratory findings on the basis of admission notes and results of investigations carried out within 30 min of admission. The list of diagnoses generated by the diagnostic tool at the ISABEL site after submission of these terms was entered in a performa. The presence of final diagnosis in the list generated by the ISABEL was the outcome measure studied. RESULTS: Records of 200 subjects (boys 111, girls 89, aged 28 days-12 years) were analyzed. Congenital heart disease, respiratory tract infections, meningitis, tetanus and septicemia were the most frequently encountered diagnoses. The diagnostic tool missed 27 diagnoses (such as septicemia, tuberculosis and seizures) in 39 subjects providing a sensitivity of 80.5%. CONCLUSION: Even without any training offered to the users, ISABEL provided a reasonable sensitivity of 80.5%. The tool holds promise of being useful in the developing countries. PMID- 16340050 TI - Effect of oil massage on growth and neurobehavior in very low birth weight preterm neonates. AB - OBJECTIVE: To study the effect of oil massage on growth and neurobehavior in preterm babies less than 1500 g. DESIGN: Randomized Controlled Trial. SETTING: Tertiary level neonatal unit of a teaching hospital. SUBJECTS: Neonates with birth weight <1500 grams, gestation >37 weeks, receiving enteral feeds of at least 100 mL/kg/day and less than 10 days of age. OUTCOME VARIABLES: Primary Weight gain 28 days after enrolment. Secondary-Neonatal neurobehavior, change in other anthropometry and serum triglycerides. METHODS: Eligible neonates were randomized to one of the three groups (a) massage with oil (b) massage without oil and (c) no massage. Weight, length, head circumference and triceps skin fold thickness were measured in the three groups at regular intervals. Serum triglyceride levels were measured at enrolment and at completion. Neurobehavior using Brazeltons Neonatal Behavior Assessment Scale (NBAS) was assessed at enrolment and after 10 days of intervention. RESULTS: Weight gain in the oil massage group (365.8 +/- 165.2g) was higher compared to the only massage group (290.0 +/- 150.2g) and no massage group (285.0 +/- 170.4g). This difference and the difference in other anthropometric parameters was not statistically significant. Serum triglycerides and neonatal neurobehavior were comparable in the three groups. CONCLUSION: Oil application may have a potential to improve weight gain among preterm very low birth weight neonates. PMID- 16340051 TI - Diagnosis and management of malaria in children: recommendations and IAP plan of action. PMID- 16340052 TI - Current strategies in the treatment of childhood Hodgkins disease. AB - Dramatic changes in the treatment of childhood Hodgkins disease have taken place during the past three decades. Contemporary combined modality treatment regimens produce durable disease-free survival in 90 to 100%of patients with early disease and in 70 to 85%of patients with advanced disease. Studies using chemotherapy alone also report high survival rates, and current studies are few to highlight the superiority of chemo-radiotherapy vs. chemotherapy alone. After the prodigious improvement achieved in response and survival rates, current strategies aim at reducing late effects of therapy, reserving more aggressive treatment modalities for patients with high risk features. PMID- 16340053 TI - Foreign body asphyxiation in children. AB - We aimed to investigate the frequency and epidemiological features of deaths due to foreign body asphyxiation (FBA) in childhood, over 1990-2003. Of the victims, 14 (63.6%) were male and 8 (36.4%) females. The mean age of the victims was 2.2 +/- 0.6 years. There were 20 (90.9%) children between 1 and 3 years, and two other cases at 2/12 and 5 years of ages. All aspirations had occurred at home. Eight (36.4%) of the victims were dead on arrival, 11 (50%) on intervention, and 3 (13.6%) after complications. Food material was the most commonly aspirated foreign body in 81.8% of the cases, nuts being the most common (50%). Food asphyxiation remains a common problem particularly in children between 1 and 3 years of age in our region. These fatal accidents can be prevented by parental education and early recognition and management of the situation. PMID- 16340054 TI - Neurodevelopmental, functional and growth status of term low birth weight infants at eighteen months. AB - This study was done to evaluate the neurodevelopmental, functional and growth status of term infants weighing 2000 g or less at 18 months, and to analyze major medical and social factors associated with an adverse neurodevelopmental and/or functional outcome. All infants were assessed for growth, audio-visual, neurological impairment, and motor and mental development using Indian modification of Bayley Scales of infant development. A detailed history was also taken. Term infants with birth weight of >2500 g without any antenatal or neonatal complications served as controls. Fifty low birth weight (LBW) term infants and 30 controls were evaluated. The mean mental development Quotient for LBW infants [91.51(16.97)] was significantly lower than that of Controls [102.02(8.4)]; the mean motor development Quotient however was comparable. The LBW infants were significantly lagging in terms of weight, length and head circumference at assessment. Neonatal complications were associated with an abnormal motor outcome while lower Socio-economic status and maternal education were related to adverse mental status. We concluded that Term LBW infants are at a significant disadvantage in terms of growth and mental scores at 18 months. PMID- 16340055 TI - Antioxidant status in children with homozygous thalassemia. AB - The status of enzymatic and non-enzymatic anti-oxidants was evaluated in 41 patients with transfusion dependent beta-thalassemia. An additional 20 age matched children, with non-hemolytic anemia, served as controls. Fresh blood samples, obtained in the morning, were processed immediately. Plasma was stored at -80 degrees C. Levels of vitamins A and E were assayed simultaneously by HPLC. RBC vitamin A was not measurable in 29 (70.7%) thalassemics and in all the controls. Plasma vitamin A levels were lower in thalassemics than in controls (p<0.05). Vitamin E in RBCs was not measurable in 13 (31. 7%) cases. The mean level of RBC vitamin E was 3 times lower in thalassemics. Similarly, SOD enzyme activity in thalassemics, was at least 1.5 lower in comparison to the activity documented in controls (p<0. 05). The observations indicate that thalassemics have enhanced oxidative stress. Administration of selective antioxidants and a balanced diet may preclude oxidative damage. PMID- 16340056 TI - Extensive lichen sclerosus et atrophicus. PMID- 16340057 TI - Forme fruste choledochal cyst. AB - Two girls presenting with features of pancreatitis were diagnosed to have minimal dilatation of extra hepatic biliary duct (EHBD) associated with pancreatico biliary maljunction (PBMJ). A high degree of suspicion is required to diagnose this condition that has been termed Forme fruste choledochal cyst (FFCC). Both did well with pancreatico-biliary disconnection procedure and reconstruction of biliary channel using enteric conduit. PMID- 16340058 TI - Neonatal lower extremity gangrene. AB - Four neonates suffering from bilateral lower limb gangrene were referred to us for further management. Two neonates had no contributory etiology. All four received appropriate treatment thus avoiding mortality but morbidity could not be avoided. All four neonates recovered uneventfully. PMID- 16340059 TI - Oral alendronate in osteogenesis imperfecta. AB - We report a case of Osteogenesis Imperfecta (OI) in an eight-year-old boy who was admitted with complaints of recurrent long bone fractures. With oral alendronate treatment significant increment occurred in the bone mineral density and the number of fractures decreased. The usage of oral bisphosphonates is inexpensive and easy to administer in selected cases of OI. This case report supports the usage of oral alendronate treatment as an alternative treatment in OI. PMID- 16340060 TI - Nitazoxanide. PMID- 16340061 TI - Testicular involvement in blast crisis of chronic myeloid leukemia. PMID- 16340062 TI - Bilateral sternocleidomastoid tumor in an infant. PMID- 16340063 TI - Celiac crisis. PMID- 16340064 TI - Solitary intestinal leiomyoma. PMID- 16340065 TI - Celiac disease in Chennai. PMID- 16340066 TI - Wilson's disease presenting as depressive disorder. PMID- 16340067 TI - A comparative radiological assessment of five European biosphere systems in the context of potential contamination of well water from the hypothetical disposal of radioactive waste. AB - In the framework of the BioMoSA project for the development of biosphere assessment models for radioactive waste disposal the Reference Biosphere Methodology developed in the IAEA programme BIOMASS was applied to five locations, situated in different European countries. Specific biosphere models were applied to assess the hypothetical contamination of a range of agricultural and environmental pathways and the dose to individuals, following contamination of well water. The results of these site-specific models developed by the different BioMoSA partners, and the individual normalised dose to the exposure groups were compared against each other. Ingestion of drinking water, fruit and vegetables were found to be among the most important pathways for almost all radionuclides. Stochastic calculations revealed that consumption habits, transfer factors, irrigation rates and distribution coefficients (Kd(s)) were the most important parameters that influence the end results. Variations in the confidence intervals were found to be higher for sorbing elements (e.g. (36)Cl, (237)Np, (99)Tc, (238)U, (129)I) than for mobile elements (e.g. (226)Ra, (79)Se, (135)Cs, (231)Pa, (239)Pu). The influence of daughter products, for which the distribution into the biosphere was calculated individually, was also shown to be important. This paper gives a brief overview of the deterministic and stochastic modelling results and the parameter sensitivity. A screening methodology was introduced to identify the most important pathways, simplify a generic biosphere tool and refine the existing models. PMID- 16340068 TI - Age-at-exposure effects on risk estimates for non-cancer mortality in the Japanese atomic bomb survivors. AB - Statistically significant increases in non-cancer disease mortality with radiation dose have been observed among survivors of the atomic bombings of Hiroshima and Nagasaki. The increasing trends arise particularly for diseases of the circulatory, digestive, and respiratory systems. Rates for survivors exposed to a dose of 1 Sv are elevated by about 10%, a smaller relative increase than that for cancer. The aetiology of this increased risk is not yet understood. Neither animal nor human studies have found clear evidence for excess non-cancer mortality at the lower range of doses received by A-bomb survivors. In this paper, we examine the age and time patterns of excess risks in the A-bomb survivors. The results suggest that the excess relative risk of non-cancer disease mortality might be highest for exposure at ages 30-49 years, and that those exposed at ages 0-29 years might have a very low excess relative risk compared with those exposed at older ages. The differences in excess relative risk for different age-at-exposure groups imply that the dose response relationships for non-cancer disease mortality need to be modelled with adjustment for age-at-exposure. PMID- 16340069 TI - A respiratory model for uranium aluminide based on occupational data. AB - As part of an epidemiological study, doses from intake of radionuclides were estimated for workers employed during a 52-year period at the Rocketdyne/Atomics International facility in California. The facility was involved in a variety of research programmes, including nuclear fuel fabrication, spent nuclear fuel decladding, and reactor operation and disassembly. Most of the documented intakes involved inhalation of enriched uranium (U), fission products, or plutonium (Pu). Highest doses were estimated for a group of workers exposed to airborne uranium aluminide (UAl(x)) during the fabrication of reactor fuel plates. Much of the exposure to UAl(x) occurred early in the fuel fabrication programme, before it was recognised that intake and lung retention were being underestimated from urinary data due to an unexpected delayed dissolution of the inhaled material. In workers who had been removed from exposure, the rate of urinary excretion of U increased for a few months, peaked, and then declined at a rate consistent with moderately soluble material. This pattern differs markedly from the monotonically decreasing absorption rates represented by the default absorption types in the Human Respiratory Tract Model (HRTM) of the International Commission on Radiological Protection (ICRP). This paper summarises the findings on the behaviour of UAl(x) in these workers and describes material-specific parameter values of the HRTM based on this information. PMID- 16340070 TI - Action levels for automatic gamma-measurements based on probabilistic radionuclide transport calculations. AB - In the event of a nuclear or radiological emergency resulting in an atmospheric release of radioactive materials, stationary gamma-measurements, for example obtained from distributed, automatic monitoring stations, may provide a first assessment of exposures resulting from airborne and deposited activity. Decisions on the introduction of countermeasures for the protection of the public can be based on such off-site gamma measurements. A methodology is presented for calculation of gamma-radiation action levels for the introduction of specific countermeasures, based on probabilistic modelling of the dispersion of radionuclides and the radiation exposure. The methodology is applied to a nuclear accident situation with long-range atmospheric dispersion of radionuclides, and action levels of dose rate measured by a network of monitoring stations are estimated for sheltering and foodstuff restrictions. It is concluded that the methodology is applicable to all emergency countermeasures following a nuclear accident but measurable quantities other than ambient dose equivalent rate are needed for decisions on the introduction of foodstuff countermeasures. PMID- 16340071 TI - Terrestrial gamma radiation dose study to determine the baseline for environmental radiological health practices in Melaka state, Malaysia. AB - Environmental terrestrial gamma radiation dose rates were measured throughout Melaka, Malaysia, over a period of two years, with the objective of establishing baseline data on the background radiation level. Results obtained are shown in tabular, graphic and cartographic form. The values of terrestrial gamma radiation dose rate vary significantly over different soil types and for different underlying geological characteristics present in the study area. The values ranged from 54 +/- 5 to 378 +/- 38 nGy h(-1). The highest terrestrial gamma dose rates were measured over soil types of granitic origin and in areas with underlying geological characteristics of an acid intrusive (undifferentiated) type. An isodose map of terrestrial gamma dose rate in Melaka was drawn by using the GIS application 'Arc View'. This was based on data collected using a NaI(Tl) scintillation detector survey meter. The measurements were taken at 542 locations. Three small 'hot spots' were found where the dose rates were more than 350 nGy h(-1). The mean dose rates in the main population areas in the mukims (parishes) of Bukit Katil, Sungai Udang, Batu Berendam, Bukit Baru and Bandar Melaka were 154 +/- 15, 161 +/- 16, 160 +/- 16, 175 +/- 18 and 176 +/- 18 nGy h( 1), respectively. The population-weighted mean dose rate throughout Melaka state is 172 +/- 17 nGy h(-1). This is lower than the geographical mean dose rate of 183 +/- 54 nGy h(-1). The lower value arises from the fact that most of the population lives in the central area of the state where the lithology is dominated by sedimentary rocks consisting of shale, mudstone, phyllite, slate, hornfels, sandstone and schist of Devonian origin which have lower associated dose rates. The mean annual effective dose to the population from outdoor terrestrial gamma radiation was estimated to be 0.21 mSv. This value is higher than the world average of 0.07 mSv. PMID- 16340072 TI - Recent results from a study of thorium lung burdens and health effects among miners in China. AB - The purpose of this study was to obtain more information about health effects among the dust-exposed male miners of Bayun Obo Rare-Earth and Iron Mine, China. From 2,390 male miners from the seven dust-generating workshops of the mine, 136 dust-exposed miners were randomly selected for study. Of these, 64 men were from the high-dust-generating workshop and 72 from the lower-dust-generating workshops; the latter group was used as an internal control. Physical measurements and medical examinations were carried out on each of these 136 men. The average measured thorium lung burden for the high-dust-exposure miners was significantly greater than that for the group of lower-exposure miners, and the incidence of severe breathlessness and pneumoconiosis of stage 0(+) was also significantly raised in the high-exposure group relative to the low-exposure group. An epidemiological study of lung cancer mortality among all the miners and staff of this mine was also carried out. This showed significantly raised levels of lung cancer mortality in both exposed miners and unexposed workers when compared with the Chinese population, and the level in exposed miners was significantly higher than that in unexposed men. The general high rate of lung cancer mortality in the workers of the mine is attributed to high levels of cigarette smoking, and the raised rate in the exposed miners relative to the unexposed workers to inhalation of silica- and thorium-bearing dusts and thoron progeny. PMID- 16340073 TI - Investigation of occupational radiation exposures to NORM at an Irish peat-fired power station and potential use of peat fly ash by the construction industry. AB - Annually, approximately 15% of Ireland's electricity requirement is provided through the combustion of 3 x 10(6) tonnes of peat. While the literature on coal fired power generation is quite abundant, studies on the peat-fired power generation industry from a radiological point of view are scarce. A study of the largest Irish peat-fired power plant was initiated to review the potential occupational radiation exposures arising from the occurrence of naturally occurring radioactive material (NORM) at different stages of the industrial process and to investigate any radiological health consequences that may arise should peat fly ash be used as a component of building materials. Ambient gamma dose rate measurements, radon measurements, quantification of the occupational exposure from inhalation of airborne particles (personal air sampling) and gamma spectrometry analysis of peat, peat ash and effluent samples from the ash ponds were undertaken. The results indicate that the radiation dose received by any worker involved in the processing of the peat and the handling of the ash resulting from peat combustion does not exceed 150 microSv per annum. Regulatory control of the peat-fired power generation is therefore unnecessary according to the Irish legislation with regards to NORM. The potential use of peat fly ash as a by-product in the building industry was also found to have a negligible radiological impact for construction workers and for members of the public. PMID- 16340074 TI - The development of the UK radon programme. AB - The natural radioactive gas, radon, is responsible for the largest component of the radiation dose received by the average UK citizen. The risks of exposure to radon have been demonstrated and quantified in epidemiological studies of those exposed at work and in the home. In the UK, measures are in place to identify and help control the exposures in those houses where levels are highest, to limit levels in new buildings and to control exposures in the workplace. This paper outlines the development of the programme, with special reference to the identification and remediation of homes with high radon levels. PMID- 16340075 TI - Review of the radiation exposure of the UK population. AB - Since 1974 the National Radiological Protection Board (now the Radiation Protection Division of the Health Protection Agency) has produced reviews of the levels of exposure to ionising radiation in the UK, from sources of natural and artificial origin. The latest review (Watson et al 2005 Ionising Radiation Exposure of the UK Population: 2005 Review HPA-RPD-001 (Chilton: HPA-RPD)) in the series gives estimates of annual doses based predominantly on data collected for the years 2001-2003. The overall average annual dose is rounded to 2.7 mSv, and the average annual dose from natural radiation is found to be 2.2 mSv. The overall average annual dose is slightly increased over that found in the previous, 1999, review. This increase is mainly due to a larger contribution from medical irradiation. PMID- 16340076 TI - A case for nuclear generated electricity. PMID- 16340078 TI - IAEA conference on the safety of radioactive waste disposal. PMID- 16340077 TI - The Chernobyl Forum completed. PMID- 16340080 TI - Tau-induced neurodegeneration: a clue to its mechanism. PMID- 16340081 TI - Using proteomics and network analysis to elucidate the consequences of synaptic protein oxidation in a PS1 + AbetaPP mouse model of Alzheimer's disease. AB - Increasing evidence suggests that oxidative injury is involved in the pathogenesis of many age-related neurodegenerative disorders, including Alzheimer's disease (AD). Identifying the protein targets of oxidative stress is critical to determine which proteins may be responsible for the neuronal impairments and subsequent cell death that occurs in AD. In this study, we have applied a high-throughput shotgun proteomic approach to identify the targets of protein carbonylation in both aged and PS1 + AbetaPP transgenic mice. However, because of the inherent difficulties associated with proteomic database searching algorithms, several newly developed bioinformatic tools were implemented to ascertain a probability-based discernment between correct protein assignments and false identifications to improve the accuracy of protein identification. Assigning a probability to each identified peptide/protein allows one to objectively monitor the expression and relative abundance of particular proteins from diverse samples, including tissue from transgenic mice of mixed genetic backgrounds. This robust bioinformatic approach also permits the comparison of proteomic data generated by different laboratories since it is instrument- and database-independent. Applying these statistical models to our initial studies, we detected a total of 117 oxidatively modified (carbonylated) proteins, 59 of which were specifically associated with PS1 + AbetaPP mice. Pathways and network component analyses suggest that there are three major protein networks that could be potentially altered in PS1 + AbetaPP mice as a result of oxidative modifications. These pathways are 1) iNOS-integrin signaling pathway, 2) CRE/CBP transcription regulation and 3) rab-lyst vesicular trafficking. We believe the results of these studies will help establish an initial AD database of oxidatively modified proteins and provide a foundation for the design of future hypothesis driven research in the areas of aging and neurodegeneration. PMID- 16340082 TI - Plasma levels of insulin and amyloid beta 42 are correlated in patients with amnestic Mild Cognitive Impairment. AB - Epidemiological and experimental data suggest that type 2 diabetes (DM2) and sporadic late-onset Alzheimer's disease (AD) share a common mechanism, that is able to produce accumulation of insulin and amyloid beta 42 (Abeta42), the major pathogenic events respectively of the two conditions. In 71 non diabetic patients with amnestic mild cognitive impairment we found a significant linear correlation between fasting plasma levels of insulin and Abeta42 (R = +0.25, P < 0.05). The levels of both peptides were elevated in comparison to 48 age-matched cognitively normal controls. The correlation of insulin and Abeta42 plasma levels suggests a pathogenic link between DM2 and sporadic AD. PMID- 16340083 TI - Insulin and insulin-like growth factor expression and function deteriorate with progression of Alzheimer's disease: link to brain reductions in acetylcholine. AB - Reduced glucose utilization and energy metabolism occur early in the course of Alzheimer's disease (AD) and correlate with impaired cognition. Glucose utilization and energy metabolism are regulated by insulin and insulin-like growth factor I (IGF-I), and correspondingly, studies have shown that cognitive impairment may be improved by glucose or insulin administration. Recently, we demonstrated significantly reduced levels of insulin and IGF-I polypeptide genes and their corresponding receptors in advanced AD relative to aged control brains. The abnormalities in gene expression were accompanied by impaired survival signaling downstream through PI3 kinase-Akt. The present work characterizes the abnormalities in insulin and IGF gene expression and receptor binding in brains with different Braak stage severities of AD. Realtime quantitative RT-PCR analysis of frontal lobe tissue demonstrated that increasing AD Braak Stage was associated with progressively reduced levels of mRNA corresponding to insulin, IGF-I, and IGF-II polypeptides and their receptors, tau, which is regulated by insulin and IGF-I, and the Hu D neuronal RNA binding protein. In contrast, progressively increased levels of amyloid beta protein precursor (AbetaPP), glial fibrillary acidic protein, and the IBA1/AIF1 microglial mRNA transcripts were detected with increasing AD Braak Stage. Impairments in growth factor and growth factor receptor expression and function were associated with increasing AD Braak stage dependent reductions in insulin, IGF-I, and IGF-II receptor binding, ATP levels, and choline acetyltransferase (ChAT) expression. Further studies demonstrated that: 1) ChAT expression increases with insulin or IGF-I stimulation; 2) ChAT is expressed in insulin and IGF-I receptor-positive cortical neurons; and 3) ChAT co-localization in insulin or IGF-I receptor-positive neurons is reduced in AD. Together, these data provide further evidence that AD represents a neuro-endocrine disorder that resembles a unique form of diabetes mellitus (? Type 3) and progresses with severity of neurodegeneration. PMID- 16340084 TI - Plasma Abeta42 correlates positively with increased body fat in healthy individuals. AB - Obesity and overweight, well known risk factors for cardiovascular disease and type 2 diabetes, are now associated with Alzheimer's disease (AD). It remains to be determined if obesity and overweight contribute to the risk of developing AD through modulating levels of amyloid-beta (Abeta), a key molecule in AD pathogenesis. Thus, we investigated whether there were any associations between plasma Abeta levels and body mass index (BMI) or fat mass (FM) in a group of 18 healthy adults. A statistically significant correlation was found between BMI, FM, and plasma levels of Abeta42 (BMI r = 0.602, P = 0.008; FM r = 0.547, P = 0.019), the longer, more pathogenic form of Abeta, but not with plasma levels of the shorter, less pathogenic Abeta40. Although not significant, positive correlations between plasma levels of Abeta42 and levels of insulin and the inflammatory marker C-reactive protein (CRP), along with an inverse trend between plasma Abeta42 levels and levels of high density lipoprotein (HDL) were answered. These results suggest that proteins implicated in inflammation, cardiovascular disease and type 2 diabetes, which in turn are risk factors for AD, may contribute to the associations between BMI/FM and plasma Abeta42 levels. Longitudinal studies involving larger cohorts are required to determine if elevated body fat may predispose individuals to AD through increasing Abeta42 levels throughout early to late adulthood. PMID- 16340085 TI - Apple juice concentrate prevents oxidative damage and impaired maze performance in aged mice. AB - Oxidative stress contributes to age-related cognitive decline. In some instances, consumption of fruits and vegetables rich in antioxidant can provide superior protection than supplementation with purified antioxidants. Our prior studies have shown that supplementation with apple juice concentrate (AJC) alleviates oxidative damage and cognitive decline in a transgenic murine model compromised in endogenous antioxidant potential when challenged with a vitamin-deficient, oxidative stress-promoting diet. Herein, we demonstrate that AJC, administered in drinking water, is neuroprotective in normal, aged mice. Normal mice aged either 9-10 months or 2-2.5 years were maintained for 1 month on a complete diet or a diet lacking folate and vitamin E and containing iron as a pro-oxidant, after which oxidative damage was assayed by thiobarbituric acid-reactive substances and cognitive decline as assayed by performance in a standard Y-maze. Mice 9-12 months of age were unaffected by the deficient diet, while older mice demonstrated statistically-increased oxidative damage and poorer performance in a Y maze test. Supplementation with AJC prevented these neurodegenerative effects. These data are consistent with normal aged individuals being susceptible to neurodegeneration following dietary compromise such as folate deficiency, and a hastened onset of neurodegeneration in those individuals harboring a genetic risk factor such as ApoE deficiency. These findings also support the efficacy of antioxidant supplementation, including consumption of antioxidant-rich foods such as apples, in preventing the decline in cognitive performance that accompanies normal aging. PMID- 16340086 TI - Exacerbation of Lewy bodies dementia due to memantine. AB - INTRODUCTION: Lewy body dementia (DLB) is common but frequently misdiagnosed as Alzheimer's disease plus delirium or parkinsonism. DRUGS USED IN THIS DISORDER CAN CAUSE EXACERBATIONS: neuroleptic medication is relatively contraindicated because some patients show severe neuroleptic sensitivity, antiparkinsonian medication has the potential to exacerbate psychotic symptoms, and even cholinesterase inhibitors, while relatively safe, have provoked adverse responses in some DLB patients. There are few data available about the use of memantine in DLB. CASE PRESENTATION: A 74-year-old man was diagnosed with Alzheimer disease and parkinsonism. After memantine was started he developed severe fluctuations in awareness, visual hallucinations, agitation, and worsened parkinsonism. When he was evaluated thoroughly, the diagnosis was revised to Lewy body dementia, leading to changes in treatment that were associated with dramatic improvement in the patient's mental status. CONCLUSIONS: In our patient, motor and cognitive symptoms worsened with memantine treatment; these resolved after memantine was discontinued. PMID- 16340087 TI - APOE2 and consanguinity: a risky combination for Alzheimer's disease. AB - OBJECTIVE: To investigate the association between APOE genotypes and Alzheimer's disease (AD) in elderly Indian subjects. The study also aims at the identification of consanguinity as disease risk factor for AD. METHODS: A total of 100 Indian patients (26 consanguineous, 74 non-consanguineous), meeting criteria for probable or definite AD and 36 cognitively healthy, elderly unrelated control subjects (spouses), were included in the study. The APOE genotyping and statistical analyses (SPSS 7.5) was performed to determine the odds of AD according to APOE genotype. RESULTS: The analysis revealed increased prevalence of E4 allele in patients as compared with controls. The difference in prevalence of E3 and E4 alleles was found to be statistically significant between consanguineous patients and controls as well as non-consanguineous patients and controls. Compared to individuals with the APOE3/3, the odds of having AD were significantly increased among those with one or more copies of the E4 allele, even after adjusting with age and sex. An interesting outcome of the study is the higher prevalence of the E2 allele in consanguineous AD patients (in contrast to previously reported studies). The risk for AD was higher in consanguineous individuals with E2/4 genotype (1.62 fold), as compared to non-consanguineous individuals. DISCUSSION: The Results support that the APOE4 allele plays a role as a risk factor for AD and suggests that the APOE2 allele may not play a protective role in the development of AD in Indians, especially individuals with family history of consanguinity. The study hypothesises that the consanguinity modifies the disease risk associated with E2 allele, although this allele is considered protective. The less number of subjects from a broader population limits the study. The wide-ranging population and ethnicity-based studies in the most inbred groups of the world may provide comprehensive insight into this conclusion. PMID- 16340088 TI - Appearance of amyloid beta-like substances and delayed-type apoptosis in rat hippocampus CA1 region through aging and oxidative stress. AB - To elucidate whether oxidative stress induces cognitive deficit, and whether nerve cells in the hippocampus, which modulates learning and memory functions in the brain, are damaged by oxidative stress and during aging, the influence of hyperoxia as oxidative stress on either the cognitive function of rats or the oxidative damage of nerve cells was investigated. Young rats showed better learning ability than both old rats and vitamin E-deficient young rats. Vitamin E supplemented young rats showed similar ability to young control rats. After they learned the location of the platform in the Morris water maze test, the young rats and vitamin E-supplemented young rats were subjected to oxidative stress for 48 h, and the old rats and vitamin E-deficient young rats were kept in normal atmosphere. The memory function of the old rats and vitamin E-deficient young rats declined even when they were not subjected to oxidative stress for 48 h. In contrast, the young rats maintained their memory function for 4 days after the oxidative stress. However, their learning abilities suddenly declined toward that of the normal old rats after 5 days. At this point, nerve cell loss and apoptosis were observed in the hippocampal CA 1 region of young rats. Vitamin E supplementation in the young rats prevented either memory deficit or the induction of delayed-type apoptosis. The old rats and vitamin E-deficient young rats kept in normal atmosphere for 48 h also showed apoptosis in the hippocampus. Also, 10 days after oxidative stress, amyloid beta-like substances appeared in the CA-1 region of control young rats; these substances were also observed in the CA-1 region of the old rats and vitamin E- deficient young rats. These results suggest that reactive oxygen species (ROS) generated by oxidative stress induced amyloid beta-like substances and delayed-type apoptosis in the rat hippocampus, resulting in cognitive deficit. Since amyloid beta in Alzheimer's disease characterized by cognitive deficit induces neuronal cell death, it is reasonable to consider that amyloid beta deposition in the brain may be associated with memory dysfunction. The results of this study imply that age-related hippocampal neuronal damage is prevented by vitamin E supplementation due to the antioxidant effect of vitamin E. PMID- 16340089 TI - Preliminary evidence of the short-term effectiveness of alternative treatments for low back pain. AB - In recent years, there has been increased popularity for use of alternative treatment modalities for low back pain with limited investigation into whether they are effective in returning the worker to normal functionality. The objective of this study was to investigate the short-term impact on the restoration of the functional ability of common treatment modalities including physical therapy, acupuncture, chiropractic treatment, and massage therapy. A within subjects design was used that compared a baseline assessment to three follow-up assessments: immediately following treatment, two days and seven days post treatment. Individuals suffering from low back pain were recruited from local clinics that specialize in one of the four treatment modalities. Twenty-four individuals who were suffering from low back pain and had started one of the treatment modalities were recruited into the study. Three-dimensional motions (ROM, velocity, and acceleration) during a standardized functional capacity protocol were measured prior to and during three follow-up periods. The results show the effectiveness of the treatments was dependent upon time and were predominantly in the non-sagittal motion planes (velocities and accelerations). Individual subjective pain levels also indicate all the treatment modalities were effective after the 1-week follow-up. Findings suggest those modalities primarily focused on the back musculature (acupuncture, massage, physical therapy) were more effective with respect to functional capacity restoration. The current results provide a quantitative evaluation, based upon trunk dynamics, of the effectiveness of these four treatment modalities. PMID- 16340090 TI - Integra artificial skin in use for full-thickness burn surgery: benefits or harms on patient outcome. AB - Full thickness burns usually lead to a great loss of skin and physiological instability because human skin is not only the biggest organ of our body it is the most important protection barrier as well. As natural skin has only limited options to recover, there is a great need for the development of synthetic skin in order to save patients with these high total body surface area full-thickness burns. Many artificial skin substitutes have been developed for this purpose so far. The benefits and harms of one substitute--Integra artificial skin--are discussed in a critical literature review. PMID- 16340091 TI - A fluoroscopy-based surgical navigation system for high tibial osteotomy. AB - High tibial osteotomy is a widely accepted treatment for unicompartmental osteoarthritis of the knee and other lower extremity deformities, particularly in young and active patients. However, it is generally recognized as a technically demanding procedure. The lack of intraoperative control of the mechanical axis of the affected limb often results in postoperative malalignments, which is one of the main reasons for poor long-term results. Moreover, inaccurate osteotomies, such as insufficient or excessive bone cut, or incorrect orientation of the chisel or saw blade, have been observed. A computer assisted intraoperative planning and navigation system is therefore proposed in order to address these technical problems. During operation, fluoroscopic images are acquired and anatomical landmarks are digitized; a patient-specific coordinate system is established accordingly. After the three-dimensional measurement of the deformity and interactive planning of the osteotomy plane, the deformity is corrected under navigational guidance. The proposed system has been successfully introduced into the clinical practice of surgery after encouraging laboratory evaluations, with results affirming that it is safe and accurate. PMID- 16340092 TI - GammaKnife surgery: safety and the identity of users. AB - In this study we investigated safety-related usability issues of an advanced medical technology, a radiosurgery system. We were interested in which criteria are important for users when a system's usability and safety is to be improved. The data collection was based on interviews and observations at three different sites where the Leksell GammaKnife is used. The analysis was qualitative. The main finding was that the user's identity or professional background has a significant impact both on how he or she views his or her role in the clinical setting, and on how he or she defines what improvements are necessary and general safety issues. In fact, the opinion even of users experienced in safety-related problems was highly influenced by how they related to the technology and its development. None of the users actually considered Leksell GammaKnife as lacking in safety, instead, their assessment was directed towards potential future system improvements. Our findings suggest that the importance of user identity or professional background cannot be neglected during the development of advanced technology. They also suggest that the user feedback should always be related to user background and identity in order to understand how important different issues are for particular users. PMID- 16340093 TI - Usefulness of quantitative ultrasound in evaluating structural and mechanical properties of bone: comparison of ultrasound, dual-energy X-ray absorptiometry, micro-computed tomography, and mechanical testing of human phalanges in vitro. AB - INTRODUCTION: Ultrasound studies evaluating bone tissue generally concentrate on two parameters--velocity and attenuation. This study aimed to determine whether ultrasound signal analysis techniques could provide additional information on the structural and mechanical characteristics of bone. MATERIALS AND METHODS: In vitro measurements were made on 26 left index fingers from human cadavers. Ultrasound measurements at the distal metaphysis and epiphysis; dual-energy X-ray absorptiometry of the whole phalanx; micro-computed tomography at the distal quarter of the phalanx (that is, the distal epiphysis and metaphysis), and mechanical three-point bending tests were performed. Univariate and multivariate linear regression techniques were used to analyze the results. RESULTS: The ultrasound parameters, speed of sound and ultrasound peak amplitude correlated significantly with the three micro-computed tomography measures used to describe the characteristics of mineralized material (r=0.69-0.79, p<0.05). Low frequency ultrasound correlated significantly with micro-computed tomography parameters describing inter-trabecular or marrow spaces (r=0.68-0.78, p<0.05). Comparison of ultrasound parameters with geometric characteristics showed that while speed of sound and ultrasound peak amplitude were related to the cortical area, moment of inertia, and mechanical load (r=0.57-0.83, p< 0.05), the amplitude of the fastest part of the ultrasound signal and full width at 80% maximum of the low frequency peak were related to the relative area of the medullary canal (r=0.40-0.43, p<0.05). DISCUSSION: Quantitative ultrasound may provide information on structural, material and mechanical characteristics of bone to the same extent and even better than DXA Bone Mineral Density. These results have been obtained by a complete and exhaustive use of QUS technology in situ but under clinical conditions. The ultrasound parameters, correctly used and combined, seem to be effective tools for investigating bone tissue. PMID- 16340094 TI - PKI-based secure mobile access to electronic health services and data. AB - Recent research works examine the potential employment of public-key cryptography schemes in e-health environments. In such systems, where a Public Key Infrastructure (PKI) is established beforehand, Attribute Certificates (ACs) and public key enabled protocols like TLS, can provide the appropriate mechanisms to effectively support authentication, authorization and confidentiality services. In other words, mutual trust and secure communications between all the stakeholders, namely physicians, patients and e-health service providers, can be successfully established and maintained. Furthermore, as the recently introduced mobile devices with access to computer-based patient record systems are expanding, the need of physicians and nurses to interact increasingly with such systems arises. Considering public key infrastructure requirements for mobile online health networks, this paper discusses the potential use of Attribute Certificates (ACs) in an anticipated trust model. Typical trust interactions among doctors, patients and e-health providers are presented, indicating that resourceful security mechanisms and trust control can be obtained and implemented. The application of attribute certificates to support medical mobile service provision along with the utilization of the de-facto TLS protocol to offer competent confidentiality and authorization services is also presented and evaluated through experimentation, using both the 802.11 WLAN and General Packet Radio Service (GPRS) networks. PMID- 16340095 TI - Rehabilitation and neurologic repair in Parkinson's disease. PMID- 16340096 TI - Parkinson's Disease Research Education and Clinical Centers (PADRECC): background and overview. AB - Parkinson's disease (PD) afflicts more than 1,000,000 people in the United States and over 50,000 veterans obtain their medical care for PD within the Veterans Health Care System. In an effort to improve care for this growing population of veterans suffering from PD, the Veteran's Health Administration established 6 Parkinson's disease Research, Education, and Clinical Centers (PADRECC) based on merit. These 6 centers offer state of the art diagnosis and treatment of PD and other movement disorders. The PADRECC also provide education for both the professional community and patients not only at the 6 sites, but also throughout the VA system through the development of a national consortium. Improving veterans' health care through research is also a priority for the PADRECC. All 6 PADRECC are participating in the largest surgical trial for the treatment of PD ever performed. Heath service researchers have already identified quality of care indicators that are now being used to evaluate care for veterans with PD. Basic researchers at the PADRECC are studying the cause of PD and are developing new therapies including stem cells. The development of the PADRECC has created an important infrastructure and attracted expertise into the VA system. They have already made great improvements in caring for veterans with PD and promises to push the field further with their research efforts. PMID- 16340097 TI - Efficacy of a multidisciplinary treatment program on one-year outcomes of individuals with Parkinson's disease. AB - The efficacy of the multidisciplinary treatment approach to the management of Parkinson's disease (PD) was examined at a regional Veteran's Administration Parkinson's Disease Research, Education and Clinical Center (PADRECC). The records of 43 consecutive individuals with PD were examined. The Unified Parkinson's Disease Rating Scale (UPDRS) was employed to assess disease progression. Changes between initial and one-year follow-up UPDRS motor functioning (Part III) scores were compared to expected disease progression from prior research. In this cohort, thirty patients (69.8%) had improved, 2 were unchanged (4.7%) and 11 patients (25.6%) had worsened at the mean 12.2-month follow-up period. The range of multidisciplinary interventions included neurology (100%), physiatrist (93%), and psychology (41.9%) visits, medication changes (60.5%), rehabilitation therapy (62.8%), functional diagnostic testing (16.3%), support group (9.3%), home exercise programs (86%), and disease and wellness education (83.7%). Statistical analyses of the individual components of the program did not demonstrate significant differences between improvers and non improvers. Clinical implications and study limitations are discussed. PMID- 16340098 TI - Falling risk factors in Parkinson's disease. AB - OBJECTIVE: To identify falling risk factors that are potentially modifiable among individuals who have idiopathic Parkinson's disease. DESIGN: A between group comparison of 19 fallers and 21 nonfallers who have Parkinson's disease, across an array of variables that have been identified as falling risk factors among the elderly and among those who have Parkinson's disease. RESULTS: Several variables were demonstrated significantly to distinguish fallers: disease duration and severity; dyskinesias associated with the use of dopaminergic agents; freezing; postural instability; depression; fear of falling; impaired fine motor control and motor planning in the feet; decreased proximal strength and muscular endurance in the legs; and a higher level of disability. CONCLUSIONS: Several of these variables can be viewed a potentially modifiable during a future intervention trial that aims to reduce falls in those who have Parkinson's disease using multidimensional risk factor modification. PMID- 16340099 TI - Gait and step training to reduce falls in Parkinson's disease. AB - INTRODUCTION: Frequent falls and risk of injury are evident in individuals with Parkinson's disease (PD) as the disease progresses. There have been no reports of any interventions that reduce the incidence of falls in idiopathic PD. PURPOSE: Assess the benefit of gait and step perturbation training in individuals with PD. DESIGN: Randomized, controlled trial. SETTING: Outpatient research, education and clinical center in a tertiary care Veterans Affairs Medical Center. OUTCOME MEASURES: Gait parameters, 5-step test, report of falls. SUBJECTS: Eighteen men with idiopathic PD in stage 2 or 3 of the Hoehn and Yahr staging. METHODS: Subjects were randomly assigned to a trained or control group. They were asked about any falls 2 weeks prior to and after an 8 week period. Gait speed, cadence, and step length were tested on an instrumented walkway. Subjects were timed while stepping onto and back down from an 8.8 cm step for 5 consecutive steps. Gait training consisted of walking on a treadmill at a speed greater than over ground walking speed while walking in 4 directions and while supported in a harness for safety. Step training consisted of suddenly turning the treadmill on and off while the subject stood in the safety harness facing either forwards, backwards, or sideways. Training occurred 1 hour per day, three times per week for 8 weeks. A two-factor (time and group) analysis of variance with repeated measures was used to compare the groups. RESULTS: Substantial reduction occurred in falls in the trained group, but not in the control group. Gait speed increased in the trained group from 1.28+/-0.33 meters/sec to 1.45+/-0.37 meters/sec, but not in the control group (from 1.26 to 1.27 m/s). The cadence increased for both groups: from 112.8 to 120.3 steps/min for the trained group and 117.7 to 124.3 steps/min for the control group. Stride lengths increased for the trained group, but not the control group. The 5-step test speed increased in the trained group from 0.40+/-0.08 steps/sec to 0.51+/-0.12 steps/sec, and in the control group (0.36+/ 0.11 steps/sec to 0.42+/-0.11 steps/sec). CONCLUSION: Gait and step perturbation training resulted in a reduction in falls and improvements in gait and dynamic balance. This is a promising approach to reduce falls for patients with PD. PMID- 16340100 TI - A brief neuropsychological protocol for assessing patients with Parkinson's disease. AB - Mild to severe cognitive impairment and depression are common among patients with Parkinson's disease (PD) and play a significant role in disability. Effective treatment planning requires a holistic assessment of cognitive and emotional difficulties. A brief neuropsychological protocol was developed to examine the pattern of cognitive and emotional functioning of patients with idiopathic PD. Data was retrospectively analyzed for the first 61 male veterans who were evaluated at an interdisciplinary movement disorders clinic. The most frequently observed cognitive impairments were in the areas of auditory learning, delayed recall memory, and visual motor functioning that required alternating tracking and sequencing. Executive dysfunction was marked by the inability of 60% of the sample to complete the Wisconsin Card Sorting Test, a measure of non-verbal reasoning with a cognitive flexibility component. Clinical interviews revealed that 28% of the sample met DSM-IV Criterion A for a Major Depressive Episode. The findings suggested that this brief neuropsychological protocol established cognitive and emotional profiles of patients with PD, consistent with other samples of patients with Parkinson's Disease. Clinical uses of the brief neuropsychological protocol are detailed. Selection of an alternative executive function measure with a lower "floor" that minimizes patient fatigue and frustration is recommended. PMID- 16340101 TI - Speech treatment for Parkinson's disease. AB - Researchers estimate that 89% of people with Parkinson's disease (PD) have a speech or voice disorder including disorders of laryngeal, respiratory, and articulatory function. Despite the high incidence of speech and voice impairment, studies suggest that only 3-4% of people with PD receive speech treatment. The authors review the literature on the characteristics and features of speech and voice disorders in people with PD, the types of treatment techniques available, including medical, surgical, and behavioral therapies, and provide recommendations for the current efficacy of treatment interventions and directions of future research. PMID- 16340102 TI - Deep brain stimulation for the treatment of Parkinson's disease: overview and impact on gait and mobility. AB - Abnormality in gait is a cardinal feature of Parkinson's disease. Walking is characterized by relatively preserved sequencing of trunk and limb movements, but diminished velocity, shortened stride length, increased base, and diminished double stance support time. The principle problem producing the gait abnormalities is dopamine deficiency, which is hypothesized to disrupt pallido thalamic modulation of cortical motor regions that automatically regulate walking. Deep brain stimulation currently is directed at either the globus pallidum internus (GPi) or subthalamic nucleus (STN) and improves many of the abnormal characteristics of parkinsonian gait with efficacy similar to dopamine replacement. The optimal target for stimulation remains uncertain and is currently being addressed in a large VA cooperative study. Our studies show that unilateral stimulation of GPi or STN improves gait to a similar extent. Functional and quantitative gait analyses confirm sustained improvement in gait dynamics with bilateral stimulation for periods for more than several years. Parkinsonian gait is also improved with rehabilitation training, primarily using external visual or auditory cues. The combination of deep brain stimulation, pharmacotherapy, and rehabilitation training may result in more effective comprehensive approaches to the reduced mobility associated with Parkinson's disease. PMID- 16340103 TI - Cell replacement efforts to repair neuronal injury: a potential paradigm for the treatment of Parkinson's disease. AB - Much has been learned from recent clinical trials exploring cell transplantation as a means to treat Parkinson's disease. Additionally, much information is being gathered in the science arena on the method of cultivation and expansion of neural stem/progenitor cells as well as catheter and cell delivery methodology. Cell replacement remains a potential promising treatment option for Parkinson's disease, however several obstacles must be overcome in order to achieve successful outcomes in future clinical trials. Hurdles include but are not limited to the identification of a reliable method of cultivation and expansion of neural stem/progenitor cells, the optimization of methods for cell delivery and the optimization of location or locations for transplantation. PMID- 16340104 TI - Work rehabilitation in Hong Kong: a recent trend. PMID- 16340105 TI - Changes in psychosocial and work-related characteristics among Clubhouse members: a preliminary report. AB - The aim of this study was to explore the changes in work-related abilities of members after attending the Clubhouse program for 12 weeks. The extent to which the changes would contribute to members' performance on simulated work tasks was also investigated. A total of 39 people with mental illnesses, primarily schizophrenia, attending Phoenix Clubhouse participated in the study. Seventeen were newly recruited members to the Clubhouse. The other 22, who served as the control group, were existing members who had been attending the Clubhouse for more than three months. All the participants were assessed by the McCarron-dial System (Observational Emotional Inventory-Revised, McCarron Assessment of Neuromuscular Development), Cognistat, Chinese Work Personality Profile, WHO Disability Assessment Schedule and WHOQOL-Bref-HK at the baseline and 12 weeks after attending the Clubhouse program. They were required to complete a set of simulated work tasks after 12 weeks. New members showed significant improvement in emotional-coping abilities (impulsive-frustration and depression-withdrawal) and work personality (task orientation, social skills, and team work) after attending the Clubhouse program for 12 weeks. The increase in emotional-coping skills (depression-withdrawal) was found to contribute to better performance on typing tasks, whereas emotional coping (anxiety) contributed to better performance on cleansing tasks. Three months' participation in the Clubhouse program appears to have positive effects on emotional-coping abilities and work personality. The emotional-coping abilities were found to predict good performance in simulated clerical and maintenance jobs. Recommendations and suggestions for further research are discussed. PMID- 16340106 TI - Outcome evaluation of work hardening program for manual workers with work-related back injury. AB - OBJECTIVE: The purpose of this study was to investigate the effectiveness of a 12 week work hardening program designed for back injured workers. STUDY DESIGN: In this study, 32 subjects were recruited. Pre- and post-assessment results were used to measure the program effectiveness. The intensity of the work hardening program was based on the overloading training principle. Subjects were contacted by phone three months after the program completion for their work status. RESULT: The findings of this study suggested that there was a significant difference in the subjects' physical demand characteristic level before and after the work hardening program. Seventy-five percent of the subjects who completed the work hardening program were able to resume employment. CONCLUSION: The rate of returning to work for back injured workers in this study was comparable to that of other studies. Thus, this study suggests that the overloading principle should be used in designing a work hardening program to improve clients' physical function. PMID- 16340107 TI - Validation of the Chinese version of the Oswestry Disability Index. AB - This study aimed to collect evidence on the structural and substantive validity, and test-retest reliability of the Chinese version of the Oswestry Disability Index (CODI). Seventy-nine patients suffering from chronic low back pain were assessed with the CODI. The results of explorative factor analysis primarily suggested a single-factor structure with nine out of 10 items (factor loading = 0.66-0.79). The sex life item was found to load on a different factor. The Cronbach's alpha of all 10 items was 0.81 (p<0.05). When the sex life item was removed from the analysis, the alpha value was increased to 0.89 (p<0.05). The test-retest reliability was estimated based on 56 participants who completed two administrations of CODI in 48 hours. The intraclass correlation coefficient (ICC) computed for the total CODI scores was 0.86 (95% C.I. = 0.81-0.91). The reliability estimated for the item scores using Kappa statistics ranged from a high of k=0.80 for the sitting item to a low of k=0.49 for the traveling item. Kappa statistics were not available for three items. The Chinese version of the Oswestry Disability Index demonstrated satisfactory validity and test-retest reliability, and so could be considered as an appropriate instrument for assessing chronic back pain-related disability in Chinese patients in Hong Kong. Further research should address the cross-cultural and measurement issues in regard to sex life in order to further improve the test content of the instrument. PMID- 16340108 TI - A comparison of self-perceived physical and psycho-social worker profiles of people with direct work injury, chronic low back pain, and cumulative trauma. AB - This study investigated the psycho-social factors among injured workers and the influence of their psycho-social profile on the readiness of return to work. Sixty-four subjects with injuries on the upper limbs or lower limbs, low back pain, or cumulative trauma disorder were recruited. The workers' profiles (self perceived physical health, mental health, and work readiness) together with the impact and types of work injury on the workers' perceived wellness were measured. The regression model of work readiness was constructed to develop a theoretical background for predicting work readiness based on different factors. The results showed that injured workers with chronic injury such as low back pain or cumulative trauma had poorer self-perception of physical health and psycho social/mental health compared to those with direct trauma. The regression analysis further affirmed that self-perceived pain and physical functioning were significant factors influencing the readiness for returning to work. Workers with low back pain were found to have lower motivation for returning to work. Other factors such as the non-verbal intelligence of the injured workers, their anxiety level, and the support they received from family members were found to have some indirect impact on their ability to return to work, but this was not statistically proven. PMID- 16340109 TI - Barriers to returning to work for people with spinal cord injuries: a focus group study. AB - The rate of employment for people with spinal cord injuries is relatively low, especially among Asians. To help individuals with spinal cord injuries return to work or retain employment, rehabilitation professionals must understand the barriers to employment and intervene to reduce or remove them. Hence, a focus group study was administered to 16 people with spinal cord injuries to explore return to work barriers. Various ideas, beliefs, responses, and values at different stages of work resettlement were elicited from both employed and unemployed participants, and then summarized. According to the results, several modifications of existing forms of rehabilitation intervention are necessary, as are further research directions. PMID- 16340110 TI - Prediction of vocational outcome of people with brain injury after rehabilitation: a discriminant analysis. AB - Due to advancements in emergency evacuation procedures and neurosurgical techniques, more patients with brain injury (BI) can survive cerebral injury and regain a satisfactory degree of recovery through rehabilitation. Their improvement in physical and cognitive functions can facilitate their reintegration into the community, especially their return to work (RTW). Different factors were reported to affect this rate of RTW in previous studies. This retrospective study thus aimed to investigate the role of demographic data, the severity of injury, residual physical deficits, activities of daily living, and cognitive function using a stepwise discriminant function analysis (DFA). Data sets for 79 persons with BI were retrieved so as to identify predictors of their vocational outcomes upon discharge and during a follow-up period. The stepwise DFA revealed that the discharge score on the Disability Rating Scale (DRS), the attention-subtest of the Neurobehavioral Cognitive Status Examination (NCSE) or Cognistat, and pre-morbid occupation were significant predictors that correctly classified 65.8% of the subjects. PMID- 16340111 TI - Content validation on the Work Performance Rating Scale for sheltered workshop workers. AB - The purpose of this study was to revise the content of the Work Performance Rating Scale (WPRS) as a valid assessment tool for sheltered workshop workers. A focus group of 11 sheltered workshop workers was recruited to collect their opinions about the relevance of the WPRS from the service users' perspectives. A panel of 23 frontline practitioners who had experience in using various work assessment instruments provided at their work setting was recruited to review the content of the WPRS. A 14-item amended version of the WPRS with five new items added was proposed based on the opinions of service users, frontline practitioners, and the concept of the Minnesota Theory of Work Adjustment (MTWA). An expert panel of 12 professionals at the senior level from various vocational rehabilitation settings was recruited to comment on the content validity of the 14-item amended version of the WPRS. The expert panel agreed that the amended version of the WPRS was related to the concept of job satisfactoriness which could be used to assess the work performance of sheltered workshop workers. Thus, the use of the MTWA for improving the content validity of the WPRS to assess the employability of sheltered workshop workers was supported by the findings of the study. PMID- 16340112 TI - Development and pilot trial of a web-based job placement information network. AB - The purpose of this project was to develop and pilot a web-based job placement information network aiming at enhancing the work trial and job placement opportunities of people with disabilities (PWD). Efficient uses of information technology in vocational rehabilitation were suggested to help improve PWD employment opportunities and thus enable them to contribute as responsible citizens to the society. In this preliminary study, a web-based employer network was so developed to explore Hong Kong employers' needs and intentions in employing PWD. The results indicated that Hong Kong employers generally agreed to arrange work trials for PWD whose work abilities match job requirements. They also expressed that they would offer permanent job placements to those PWD who showed satisfactory performance in work trials. The present study evidenced that using an information network could expedite communications between employers and job placement services, and thus job placement service outcomes. It is hoped that a job placement databank could thus be developed through accumulating responses from potential employers. PMID- 16340113 TI - Factors affecting vocational outcomes of people with chronic illness participating in a supported competitive open employment program in Hong Kong. AB - OBJECTIVES: This study aimed to analyze the ability of the Patient Retraining and Vocational Resettlement (PRAVR) program to enhance the vocational outcomes of individuals with chronic illness, and to study the socio-demographic factors associated with successful vocational outcome. STUDY DESIGN: A retrospective study of 548 individuals with various types of chronic illness who enrolled in the program between 1995 and 2003. Their socio-demographic data and their employment outcome after a six-month job skills retraining and job settlement service were collected for analysis. RESULTS: The program was found to enhance the vocational outcomes of patients who completed the program. Logistic regression identified significant factors predicting successful vocational outcomes. For the male patients, the chances of employment were higher if the onset of illness had occurred at least 10 years before (odd ratios = 0.326). For the female patients, the chances of employment were higher if they had been unemployed for less than 1 year (odd ratio = 3.8). CONCLUSIONS: The PRAVR program is able to enhance the vocational outcomes of people with chronic illness in Hong Kong. The factors which were found to relate to successful employment were unique to the local situation. Further studies should explore these factors in a more in depth manner. PMID- 16340114 TI - The working alliance as a model for interdisciplinary collaboration. AB - Rehabilitation and health care researchers are increasingly called upon to examine contemporary disability issues from an interdisciplinary perspective. Therefore, the purpose of this article is to (a) borrow from the counseling literature to introduce the working alliance, a model developed by Bordin [1] to facilitate positive therapeutic relationships; and (b) to discuss its application to interdisciplinary research. The authors, who represent the fields of nursing, rehabilitation, and health communication, describe characteristics of the working alliance and draw upon their experiences to illustrate the key components of effective working alliances (bonds, goals, and tasks). We also introduce strategies for resolving disagreements and conclude the article with a discussion regarding the multiple rewards that can be acquired from interdisciplinary research. PMID- 16340116 TI - Bone quality: summary of NIH/ASBMR meeting. PMID- 16340117 TI - Longitudinal micro-CT scans to evaluate bone architecture. PMID- 16340118 TI - Sacrificial bonds in the interfibrillar matrix of bone. PMID- 16340119 TI - Collagen glycation and its role in fracture properties of bone. PMID- 16340120 TI - Structural heterogeneity in bone: good or bad? PMID- 16340121 TI - Summary--Measuring "bone quality". AB - The idea of bone quality is well-established in the literature and represents a real conundrum in the treatment of osteoporosis. On the one hand, there are measurements for patients that predict fracture risk for the population as a whole, but between individual patients, one will fracture but another will not, despite the fact that all of the technical measurements we use to predict fracture risk are the same. There are, of course, many aspects of bone mechanical properties that cannot yet be measured in patients. The session began with a discussion of what bone quality is, then the speakers presented work on novel aspects of bone properties that could help explain why fracture prediction in vivo is inexact. PMID- 16340122 TI - Generation and function of osteocyte dendritic processes. PMID- 16340123 TI - DMP1 is essential for osteocyte formation and function. PMID- 16340124 TI - PTH and osteocytes. PMID- 16340125 TI - New observations on bone fragility with glucocorticoid treatment. Results from an in vivo animal model. PMID- 16340126 TI - Summary--Osteocytes and mechanotransduction. PMID- 16340127 TI - NFATc1 directly induces the human beta3 integrin gene in osteoclast differentiation. PMID- 16340128 TI - Wdr5, a novel WD repeat protein, regulates osteoblast and chondrocyte differentiation in vivo. PMID- 16340129 TI - Transcriptional mechanism of COMP gene expression and chondrogenesis. PMID- 16340130 TI - Bone tissue material properties are altered during osteoporosis. PMID- 16340131 TI - Does exercise during growth influence osteoporotic fracture risk later in life? PMID- 16340132 TI - Inhibition of NFAT increases osteoblast differentiation by increasing Fra-2 expression. PMID- 16340133 TI - Genetically altered mice for bone research. PMID- 16340134 TI - Viewing problems in bone biology from the perspective of lineage identification. PMID- 16340135 TI - Summary--Tutorial: Genetically modified animal models to study bone and cartilage. PMID- 16340136 TI - Selective androgen receptor modulators--prospects for emerging therapy in osteoporosis? PMID- 16340137 TI - Treatment of postmenopausal osteoporotic women with parathyroid hormone 1-84 for 18 months increases cancellous bone formation and improves cancellous architecture: a study of iliac crest biopsies using histomorphometry and micro computed tomography. PMID- 16340138 TI - Summary--Novel therapies for osteoporosis. PMID- 16340139 TI - Periosteal stem cells are essential for bone revitalization and repair. PMID- 16340140 TI - Distinct osteogenic activity of BMPs and their orthopaedic applications. PMID- 16340141 TI - Summary--Cell therapies for orthopedic applications. PMID- 16340143 TI - Proprioception, gait kinematics, and rate of loading during walking: are they related? AB - The cyclic nature of walking can lead to repetitive stress and associated complications due to the rate of loading (ROL) experienced by the body at the initial contact of the foot with the ground. An individual's gait kinematics at initial contact has been suggested to give rise to the ROL, and a repetitive, high ROL may lead to several disorders, including osteoarthritis. Additionally, proprioception, the feedback signaling of limb position and movement, may play a role in how the foot strikes the ground and thus, the ROL. Our goal was to explore the relationship between proprioception, gait kinematics and ROL. Thirty eight women were recruited for gait analysis, and the gait characteristics 50 ms prior to and at initial contact were examined. Two proprioception tests, joint angle reproduction and threshold to detect passive motion were used to examine the subject's proprioceptive acuity. Our results indicate that individuals with a larger knee angle (i.e., greater extension) 50 ms prior to initial contact (IC) experience a higher ROL during gait and have poorer proprioceptive scores. However, it remains unclear whether poor proprioception causes a high ROL or if a high ROL damages the mechanoreceptors involved in proprioception, but the apparent relationship is significant and warrants further investigation. PMID- 16340144 TI - Clinical practice guidelines proposed by the Hellenic Foundation of Osteoporosis for the management of osteoporosis based on DXA results. AB - In recent years guidelines for the testing and treatment of osteoporotic patients have been published by recognised organisations, including the World Health Organisation (WHO), the National Osteoporosis Foundation (NOF) and the International Osteoporosis Foundation (IOF). Dual Energy X-ray Absorptiometry (DXA) has been considered the technique of choice because of its excellent precision and ability to predict osteoporotic fractures. Last December, based on the Appraisal of the Guidelines for Research and Evaluation (AGREE), the Hellenic Foundation of Osteoporosis, in collaboration with other scientific societies, provided guidelines for the use of DXA for the diagnosis, monitoring and treatment of osteoporosis and Quality Assurance (QA) of these systems. According to these guidelines, the adequacy of the present number of DXA units in Greece was assessed. There are 367 DXA units in Greece, and almost 50% are located in the capital city, Athens, where 34.1% of the population lives. The distribution of DXA devices per resident in the Greek provinces (except Attica) is between 4.2 units/100,000 heads (Ionian Islands) and 1.6 units/100,000 heads (Sterea Hellas). These guidelines have resulted in a suggestive yearly repeat of the measurements, to ensure the precision of the method, but mainly for reasons of compliance. Finally, these guidelines are viewed as a work in progress and will be updated periodically in response to advances in this field. PMID- 16340146 TI - Tilt-table testing as a predictor of + GZ tolerance. AB - BACKGROUND: G-tolerance assessment is an essential element of both military pilot and pilot candidate evaluation. AIMS: Attempt at prediction of individual relaxed + Gz tolerance on the basis of head-up tilt (HUT) testing. SETTINGS AND DESIGN: In two stages, 20 healthy men at the mean age 21.5 years took part in this study. The first stage, a 45 min, HUT test was performed using the Westminster protocol. During the second stage each underwent a centrifuge evaluation in response to gradual onset rate profiles. METHODS AND MATERIAL: In each subject, heart rate (HR) and blood pressure (BP) before and at 2, 15 and 45 min of the tilt-test were recorded. The gravity-load centrifuge (GOR) studies were carried out [following the standard GOR programme, at 0.1 G%divide;s rate of gravity load increase until the gravity load tolerance limit (loss of peripheral vision) was reached]. STATISTICAL ANALYSIS USED: Relationships between variables were explored using Kendall's tau-B correlation coefficient. The critical P-level was one-tailed 0.05. RESULTS: In four of 20 subjects (20%), vasovagal syncope occurred during the tilt test. G-level tolerance of this group (of +Gz accelerations) lay in the range from +4 to +8.1 Gz, (+5.72 ? 0.86 Gz average) and was comparable to the group without syncope. Loss of consciousness did not occur in any subjects during the centrifuge test. No statistically significant correlation was observed between HR and BP during tilt test and tolerance to +Gz accelerations. CONCLUSIONS: The result of tilt testing, carried out according to the Westminster protocol, was not useful in predicting individual tolerance to +Gz gravity loads. PMID- 16340147 TI - Accurate blood pressure recording: is it difficult? AB - BACKGROUND: Blood pressure (BP) measurement is a routine procedure but errors are frequently committed during BP recording. AIMS AND SETTINGS: The aim of the study was to look at the prevalent practices in the institute regarding BP recording. The study was conducted in the Medicine Department at Government Medical College, Chandigarh, a teaching institute for MBBS students. METHODS: A prospective, observational study was performed amongst the 80 doctors in a tertiary care hospital. All of them were observed by a single observer during the act of BP recording. The observer was well versed with the guidelines issued by British Hypertension Society (BHS) and the deviations from the standard set of guidelines issued by BHS were noted. The errors were defined as deviations from these guidelines. STATISTICAL METHODS: The results were recorded as percentage of doctors committing these errors. RESULTS: In our study, 90% used mercury type sphygmomanometer. Zero error of the apparatus, hand dominance was not noted by any one. Every one used the standard BP cuff for recording BP. 70% of them did not let the patient rest before recording BP. 80% did not remove the clothing from the arm. None of them recorded BP in both arms. In out patient setting, 80% recorded blood pressure in sitting position and 14% in supine position. In all the patients where BP was recorded in sitting position BP apparatus was below the level of heart and 20% did not have their arm supported. 60% did not use palpatory method for noticing systolic BP and 70% did not raise pressure 30-40 mm Hg above the systolic level before checking the BP by auscultation. 80% lowered the BP at a rate of more than 2 mm/s and 60% rounded off the BP to nearest 5-10 mm Hg. 70% recorded BP only once and 90% of the rest re inflated the cuff without completely deflating and allowing rest before a second reading was obtained. CONCLUSION: The practice of recording BP in our hospital varies from the standard guidelines issued by the BHS. PMID- 16340148 TI - Rapid assessment survey of earthquake affected Bhuj block of Kachchh District, Gujarat, India. AB - RESEARCH QUESTIONS: How much human loss would have caused by the earthquake in Bhuj block? What is the environmental sanitation status? OBJECTIVES: (1) To assess human loss and injuries after the earthquake in Bhuj block.(2) To study the status of some relief activities.(3) To study the environmental sanitation status of the earthquake affected Bhuj block. STUDY DESIGN: Cross-sectional study. SETTINGS: Bhuj block. PARTICIPANTS: All villages excluding Bhuj city of Bhuj block. STATISTICAL ANALYSIS: Proportions, chi-square test, chi-square for trend. RESULTS: Survey was done in 144 villages; there were total 541 deaths with death rate of 3.18 per 1000 population. Death rate was significantly associated with distance of village from epicenter (chi-square for trend significant, P < 0.001). Among victims, majority were children 171 (45.4%) and women 107 (28.4%). Relief work was significantly associated with accessibility of village (P < 0.001) and few interior pockets were deprived of help. Total 56 (38.9%) villages were entirely dependent on water tanker for water supply and in 61 (42.4%) villages drinking water was used without chlorination. In 142 (98.6%) villages open-air defecation was practiced. Diseases such as URTIs, diarrheal diseases, fever and conjunctivitis were commonly observed in the field area. PMID- 16340149 TI - Cesarean scar endometriosis--report of two cases. AB - Endometriosis is presence of functioning endometrial tissue outside the uterine cavity. Endometriosis can sometimes occur in a previous surgical scar. Scar endometriosis is rare and difficult to diagnose. It mostly follows obstetrical and gynecological surgeries. This condition is often confused with other surgical conditions. We are reporting two cases of scar endometriosis following cesarean section, which were misdiagnosed as stitch granuloma initially. Medical treatment was not helpful. Both the patients required wide surgical excision of the lesion. The pathogenesis, diagnosis and treatment of this condition are being discussed. PMID- 16340150 TI - Sarcoma-like mural nodules in ovarian mucinous cystadenomas--a report of two cases. AB - Sarcoma-like mural nodule is a very rare occurrence in a mucinous tumour of the ovary. Two such nodules having morphologically benign features with osteoclastic giant cells, in the wall of mucinous cystadenoma are described. In addition, these nodules exhibited reactive vascular proliferations. The sarcoma-like nodules are associated with a favourable outcome and must be distinguished from other malignant nodules composed of sarcoma or anaplastic carcinoma that may also occur in the wall of mucinous ovarian tumours. PMID- 16340151 TI - Bilateral hemorrhagic cerebellar infarct in primary antiphospholipid syndrome. PMID- 16340152 TI - Myxoid adrenal cortical carcinoma--a rare variant of adrenocortical carcinoma. PMID- 16340153 TI - Variable neurotransmission within intrinsic cardiac ganglia. PMID- 16340154 TI - Activation of spinal anti-analgesic system following electroacupuncture stimulation in rats. AB - We evaluated the interaction between electroacupuncture (EA)-induced antinociception and an endogenous anti-analgesic system. EA was applied to the ST 36 acupoint for 45 min in male Sprague-Dawley rats, and pain thresholds were assessed by the hind-paw pressure test. EA produced a marked increase in pain thresholds and its antinociceptive action was completely reversed by naloxone (5 mg/kg). The analgesic effects of subcutaneous morphine (7 mg/kg) following EA stimulation were significantly attenuated. The attenuation of morphine analgesia was inversely proportional to the time intervals between EA termination and morphine injection, and the effect was not observed 120 min after EA stimulation. The analgesic effects of i.t. morphine (10 microg), but not i.c.v. morphine (25 microg), following EA were also attenuated. On the other hand, systemic morphine (7 mg/kg)-induced hyperthermia was not affected by EA. Moreover, i.c.v. morphine, but not i.t. morphine, produced hyperthermia. The i.c.v. morphine-induced hyperthermia was not affected by EA, similar to i.c.v. morphine analgesia. These results suggest that the attenuation of morphine analgesia following EA, that is, the activation of an endogenous anti-analgesic system, is closely related to the activation of an analgesic system by EA and that the spinal cord plays a critical role in the activation of the endogenous anti-analgesic systems. PMID- 16340155 TI - The protective effect of H2-receptor activation against the duration of myocardial hypoxia/reoxygenation-induced ventricular fibrillation in sensitized guinea-pig hearts. AB - Patients with high serum immunoglobulin E levels were reported to be protected against sudden death during acute myocardial infarction. The protection mechanism might be attributed to the facilitation of histamine release from sensitized mast cells; however, this remains to be clarified. In this study, we examined the influence of sensitization on ventricular fibrillation (VF) induced by myocardial hypoxia/reoxygenation (H/R). Guinea pigs were actively sensitized by subcutaneous injection of ovalbumin in Bordetella pertussis vaccine. Hearts isolated from non sensitized and sensitized guinea pigs were subjected to 30-min hypoxia / 30-min reoxygenation using a Langendorff apparatus. The amount of histamine released in the sensitized guinea-pig hearts was elevated, and the duration of VF was found to be reduced. The treatment with a histamine H2-receptor antagonist inhibited the reduction of VF duration. Treatment of the non-sensitized hearts with the histamine H2-receptor agonist resulted in the decrease of VF duration to the same level as that in the sensitized hearts. In conclusion, these results suggest that the risk of sudden death during myocardial H/R may be attenuated in the sensitized hearts and that histamine H2-receptor activation due to the released histamine may be involved in the protective effect. PMID- 16340156 TI - Elucidating the inhibitory mechanisms of magnolol on rat smooth muscle cell proliferation. AB - The pathological mechanism of percutaneous transluminal coronary angioplasty induced restenosis has been attributed to outgrowth of vascular smooth muscle cells. Pretreatment with antioxidants has been shown to reduce restenosis. Magnolol, an active compound of Magnolia officinalis, has exhibited approximately 1,000 times more potent antioxidant effects than alpha-tocopherol. In this study, we demonstrate, using cytometric analysis, an approximate 61% reduction of smooth muscle cells progressing to the S-phase by 0.05 mg/ml of magnolol. A BrdU incorporation assay also showed a significant reduction (73%) of DNA synthesis using 0.05 mg/ml of magnolol. The protein level of the proliferating cell nuclear antigen was suppressed by approximately 48% using 0.05 mg/ml of magnolol. This was in agreement with the promoter activity of nuclear factor-kappa B, which was also attenuated by 0.05 mg/ml of magnolol. Since receptor interacting protein and caspase-3 protein expression levels were both increased by magnolol in a dose dependent manner, the apoptotic pathway may mediate the inhibition of cell growth. Our finding that malondialdehyde formation was significantly inhibited by 0.05 mg/ml of magnolol further supported the antioxidant effect of magnolol. These studies suggest that magnolol might be a potential pharmacological reagent in preventing balloon injury-induced restenosis. PMID- 16340157 TI - Lithium: potential therapeutics against acute brain injuries and chronic neurodegenerative diseases. AB - In addition to the well-documented mood-stabilizing effects of lithium in manic depressive illness patients, recent in vitro and in vivo studies in rodents and humans have increasingly implicated that lithium can be used in the treatment of acute brain injuries (e.g., ischemia) and chronic neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, tauopathies, and Huntington's disease). Consistent with this novel view, substantial evidences suggest that depressive illness is not a mere neurochemical disease, but is linked to gray matter atrophy due to the reduced number/size of neurons and glia in brain. Importantly, neurogenesis, that is, birth/maturation of functional new neurons, continues to occur throughout the lifetime in human adult brains (e.g., hippocampus); the neurogenesis is impaired by multiple not-fully defined factors (e.g., aging, chronic stress-induced increase of glucocorticoids, and excitotoxicity), accounting for brain atrophy in patients with depressive illness and neurodegenerative diseases. Chronic treatment of lithium, in agreement with the delayed-onset of mood-stabilizing effects of lithium, up-regulates cell survival molecules (e.g., Bcl-2, cyclic AMP-responsive element binding protein, brain-derived neurotrophic factor, Grp78, Hsp70, and beta-catenin), while down regulating pro-apoptotic activities (e.g., excitotoxicity, p53, Bax, caspase, cytochrome c release, beta-amyloid peptide production, and tau hyperphosphorylation), thus preventing or even reversing neuronal cell death and neurogenesis retardation. PMID- 16340158 TI - Hepatocyte-targeted MR contrast agents: contrast enhanced detection of liver cancer in diffusely damaged liver. AB - The performance of hepatocyte-targeted magnetic resonance (MR) contrast agents in the detection of liver tumor was tested in rats with hepatitis. Hepatocyte targeted MR contrast agents (paramagnetic hepatobiliary complex [manganese-DPDP] and superparamagnetic iron oxide coated with arabinogalactan [SPIO-AG]) were injected into normal rats and rats with carbon tetrachloride-induced hepatitis. Before and after injection of either contrast agent, ex vivo relaxometry (0.94T) or in vivo MR imaging (1.0T) were performed. The obtained liver and tumor T1 and T2 relaxation times, liver and tumor signal-to-noise ratios (SNR), and tumor liver contrast-to-noise ratios (CNR) of control rats and rats with hepatitis were compared. Both relaxometry and MR imaging showed that MnDPDP and SPIO-AG selectively enhanced liver tissue in controls and in rats with hepatitis to the same degree, and little tumor enhancement was seen in either group. As a result, no significant difference between control rats and rats with hepatitis was observed in the postcontrast tumor-liver CNR. For a MnDPDP-enhanced CNR with spin echo (SE) of 310/15, the results were -10.4+/-3.6 in control rats vs. -11.5+/-1.4 in rats with hepatitis; for a SPIO-AG-enhanced CNR with SE 2000/45 and 2000/90, respectively, the results were 30.7+/-9.2 and 18.7+/-4.7 in control rats vs. 31.9+/-7.1 and 17.7+/-2.4 in rats with hepatitis. These results indicate that hepatocyte-targeted contrast agents effectively enhance liver tissue and enhance liver-tumor image contrast despite hepatocellular dysfunction. PMID- 16340159 TI - MR findings of nine cases of palatal tumor. AB - PURPOSE: To assess the magnetic resonance imaging (MRI) findings of pathologically confirmed palatal tumors. METHODS: Nine cases of palatal tumor were studied. Clinical data, MRI findings, and pathological diagnoses were evaluated. RESULTS: Five cases were tumors of the hard palate and four of the soft palate. Signal intensity on T1-weighted images varied, and hyperintensity was observed on T2-weighted images. Adenoid cystic carcinoma and diffuse large B cell lymphoma showed homogenous signal intensity. Other tumors showed heterogeneous signal intensities. On dynamic contrast analysis, malignant pleomorphic adenoma, adenoid cystic carcinoma, diffuse large B cell lymphoma, and peripheral T cell lymphoma showed early enhancement. On post-contrast T1-weighted images, hard palate pleomorphic adenoma, malignant pleomorphic adenoma, adenoid cystic carcinoma, diffuse large B cell lymphoma, and peripheral T cell lymphoma showed strong enhancement. Although the borders of the tumors were classified as clear in 6 cases treated surgically, macroscopic and microscopic borders of the tumors were unclear. Adenoid cystic carcinoma and hard palate diffuse large B cell lymphoma invaded the maxillary bone. CONCLUSION: Magnetic resonance findings of palatal tumor varied in different histologies. Even with a small palpable portion, malignant tumors could directly infiltrate surrounding structures, which demonstrated well on MRI. PMID- 16340160 TI - MT+/V5 activation without conscious motion perception: a high-field fMRI study. AB - While activity of MT+/V5 neurons is believed to be necessary for the conscious perception of visual motion, whether neural activity in MT+/V5 is a sufficient condition for the conscious perception of motion in vision still remains unanswered. A high-field (3.0 Tesla) functional magnetic resonance imaging (fMRI) study was designed and performed to answer this specific question. Eleven healthy subjects viewed a checkerboard pattern stimulus reversed in contrast at 0.2, 12, 30, and 60 Hz while being probed for activation in MT+/V5. At 0.2 Hz, all viewers perceived pattern-reversal which was stationary in position. However, at 12, 30, and 60 Hz, many subjects perceived apparent motion (e.g., vertical and horizontal flows) in the square pattern. At 12 and 30 Hz reversals, MT+/V5 was activated in all subjects (11/11). Nevertheless, three out of eleven (3/11) subjects denied motion perception in these conditions. At 60 Hz reversal, as many as seven out of eleven (7/11) subjects failed to see motion but activation in MT+/V5 was found in the majority (5/7) of the subjects. The results demonstrated that significant MT+/V5 activation occurs without accompanying subjective awareness of seeing motion in an apparent-motion stimulus, indicating that neural activity in MT+/V5 does not represent a sufficient condition for conscious perception of motion in vision. PMID- 16340162 TI - The passage to human MR microscopy: a progress report from Niigata on April 2005. PMID- 16340161 TI - An FMRI study to investigate auditory attention: a model of the cocktail party phenomenon. AB - In human life, discrimination of a target voice from other voices or sounds is indispensable, and inability for such discrimination results in sensory aphasia. To investigate the neuronal basis of the attentional system for human voices, we evaluated brain activity during listening comprehension tasks using functional magnetic resonance imaging (fMRI) at 3T. Diotic listening comprehension tasks, in which a narration was superimposed by another given by the same speaker (SV experiment) or by a different speaker (DV experiment), were presented to normal volunteers. The story indicated in the baseline task blocks, in which only one narration was presented, was intensively followed during the superimposed task blocks. In each experiment, 6 task blocks, 3 blocks for each condition, and 7 rest blocks were alternatively repeated, and the contrast of the superimposed condition to the baseline condition in each session was obtained. In the DV experiment, compared with the control condition, activation in Wernicke's area (BA22) was increased. In the SV experiment, activation in the frontal association cortex (BA6, BA9/ 46, BA32, BA13/47) was additionally increased. These results suggested that difficulty in phonological processing to discriminate human voices calls for further semantic, syntactic, and prosodic processing, as well as augmented selective attention. PMID- 16340163 TI - Alternate biplanar MR navigation for microwave ablation of liver tumors. AB - Real-time MR (magnetic resonance) images in two perpendicular planes, both of which included the path of the needle, were utilized for MR-guided microwave ablation of liver tumors. The two image planes were automatically and alternately switched by new MR scanner control software installed on an external PC. This technique is possible only with MRI (magnetic resonance imaging) units with multiplanar and multisection capabilities. Reformatted images in the corresponding two planes were also constructed from preoperative three dimensional volume data. These four images (two real-time and two reformatted) were continuously visible to the surgeons. These images enabled the needle position in the three-dimensional space to be accurately and clearly recognized, in contrast to the difficulty encountered with two-dimensional MR images in a single image plane. This technique was also applied to MR temperature mapping during microwave ablation, as it allowed monitoring of the spread of the heat in a three-dimensional space. This type of computer-integrated image navigation was demonstrated to be feasible for MR-guided microwave ablation of liver tumors. PMID- 16340164 TI - Parallel imaging of head with a dedicated multi-coil on a 0.4T open MRI. AB - BACKGROUND: Parallel imaging is widely used for cylindrical magnetic resonance imaging (MRI); however, few studies apply parallel imaging to open MRI. We previously developed a parallel method called "RAPID" (rapid acquisition through a parallel imaging design) for imaging the heart on a 0.7T open MRI apparatus, and we have now developed a RAPID head coil and shading correction algorithm for imaging the brain with a 0.4T open MRI apparatus. Images acquired with RAPID were compared with those acquired using a conventional quadrature-detection (QD) head coil. MATERIALS AND METHODS: The images were acquired using a dedicated 4-channel RF receiving coil consisting of a solenoid coil and surface coils. For MRI of the brain, we developed 2 methods to acquire the necessary calibration data: a pre scan method that acquires the calibration data before the main scans and a self calibration method that acquires the calibration data and imaging data simultaneously. We also modified the algorithm for calculating the shading distribution so that it only uses acquired image data and then corrects the shading. RESULTS: RAPID was applied for T1-weighted, T2-weighted, fluid attenuation inversion recovery (FLAIR), time-of-flight (TOF), and diffusion weighted echo-planar (DW-EPI) imaging. The RAPID images had no visible unfolded artifacts or motion artifacts. Images with the same contrast as that with a conventional QD coil were acquired using the RAPID coil and shading correction. CONCLUSION: These preliminary results show that RAPID can be applied to imaging of the head using a 0.4T open MRI apparatus. PMID- 16340165 TI - Cerebellar atrophy attributed to cerebellitis in two patients. AB - In a review of magnetic resonance (MR) images from patients with spinocerebellar degeneration (SCD), we found 2 rare patients considered to be in late-stage cerebellitis who showed isolated cerebellar atrophy. The patients were negative for the spinocerebellar ataxia (SCA) genes and had no symptoms of hypothyroidism, history of malignant tumors, or history of alcohol and drug (phenytoin) abuse, which may cause cerebellar atrophy. MR images demonstrated generalized atrophy of the cerebellum, excluding the brainstem or cerebrum. In these cases, moreover, slightly high intensities were noted in the affected cerebellar cortices on fluid attenuated inversion recovery (FLAIR) images. The distribution of widening of the folia and cortical high intensities on FLAIR images might be important clues with which to diagnose late-stage cerebellitis. PMID- 16340166 TI - Role of noradrenergic receptors in the bed nucleus of the stria terminalis in regulating pulsatile luteinizing hormone secretion in female rats. AB - The bed nucleus of the stria terminalis (BNST) is one of the brain areas densely innervated by noradrenergic neurons originating in the brain stem. The present study aims to determine the role of noradrenergic receptors in the BNST in regulating pulsatile luteinizing hormone (LH) secretion in female rats. Ovariectomized (OVX) or estrogen-primed OVX (OVX+E2) rats received three 1-h interval injections of 0.05 micromol of noradrenaline (NA), phenylephrine (alpha1 adrenergic receptor agonist), clonidine (alpha2-agonist), or isoproterenol (beta agonist) into the BNST. Injection of NA or alpha1-adrenergic agonist into the BNST strongly suppressed pulsatile LH secretion in OVX+E2 rats with a significant (P < 0.05) decrease in the mean LH level for 3 h and LH pulse frequency, but alpha2-and beta-agonists did not affect any of the LH pulse parameters. In OVX animals, alpha1- and alpha2-adrenergic agonists caused a significant change in LH pulse frequency and amplitude, respectively, though the effect was not as apparent as the NA- or alpha1-agonist-induced changes in OVX+E2 animals. These results indicate that NA inputs to the BNST suppress pulsatile LH secretion via alpha1-adrenergic receptors and that estrogen enhances this suppression. PMID- 16340167 TI - Genetically modified dendritic cells for therapeutic immunity. AB - Dendritic cells are professional antigen presenting cells, which show an extraordinary capacity to initiate primary immune responses by stimulating T cells. This established function of dendritic cells has attracted much attention in efforts to develop useful vaccines for the treatment of cancer and infectious diseases. Designing effective strategies to generate clinical dendritic cell based vaccine protocols remains a challenging field of research. The successful realization of immunotherapy utilizing dendritic cells will depend on modifications of these protocols to optimize the natural stimulatory properties of dendritic cells, such as genetic modification of dendritic cells. This review focuses on dendritic cell gene modifications for enhancing the multiple effector functions of dendritic cells, including viral and non-viral gene transfer into dendritic cells, and a variety of transferred genes, such as those encoding antigens, co-stimulatory molecules, cytokines, and chemokines. PMID- 16340168 TI - Appearance of abnormal electrocorticogram patterns during umbilical cord compression in sheep fetus. AB - A frequent cause of fetal acidemia, which sometimes results in hypoxic-ischemic encephalopathy, is umbilical cord compression associated with uterine contraction. Using a sheep model of fetal acidemia, we examined the changes in electrocorticogram (ECoG), carotid artery blood flow, arterial blood pressure and fetal heart rate during cord compression. A characteristic burst of ECoG spikes emerged during cord compression at fetal arterial pH 7.18 even before the pH went down to severe fetal acidemia (less than 7.10). The administration of a neuromuscular blocking agent to the fetus did not abolish the appearance of the spikes. These results suggest that cord compression may cause abnormal brain excitement even in the absence of severe fetal acidemia and that this abnormal excitement can lead to fetal brain dysfunction, if cord compression is repeated or prolonged. PMID- 16340169 TI - Suicide ideation after the 1999 earthquake in Marmara, Turkey. AB - This study aims to investigate the psychological effects of the earthquake. We investigated the psychological conditions of 3,609 students survived from the Marmara Earthquake, which occurred on 17 August 1999. The Beck Depression Inventory (BDI) was employed to assess the levels of depression and other psycho pathological states. The BDI was classified as mild depression if the score was < or = 13, moderate depression (14-24) and serious depression (> or = 25). Depression level was estimated as mild in 71.5% of the students, and serious depression in 9.6% of the students. The prevalence of suicidal tendency/thought was 16.7% in this study. The prospect of suicidal thought was 1.76-time (95% Confidence interval [CI]: 1.40-2.22) higher in the students who were injured or whose relatives were injured seriously enough to require medical treatment. Suicidal thought was higher by 1.57 times (95% CI: 1.28-1.92) in students who lost their relatives and by 1.35 times (95% CI: 1.13-1.63) in those who saw extensive damage or destruction occurred in their home or property. According to logistic regression analyses, the gender influenced the thought of suicide; suicide thought was 0.71 (95% CI: 0.60-0.85) time lower in females than males. The present study indicates that injury to the self or to the loved ones, damage to home or property, or the loss of family members as a result of the earthquake enhances the suicidal tendencies. PMID- 16340170 TI - A nonsense polymorphism (Y319X) of the solute carrier family 6 member 18 (SLC6A18) gene is not associated with hypertension and blood pressure in Japanese. AB - We investigated the possible association of solute carrier family 6 member 18 (SLC6A18) with hypertension and blood pressure in Japanese, since the homologous murine XT2 gene was recently reported to be associated with hypertension. The entire coding region of SLC6A18 was sequenced in 30 unrelated Japanese subjects. The deleterious effects of the observed nonsynonymous single nucleotide polymorphisms (SNPs) on the phenotype were predicted using bioinformatics software. We tested the associations of one deleterious SNP (Y319X) with blood pressure and hypertension in a general population of 1,004 subjects in one area of Japan. Both quantitative and qualitative analyses adjusting for age and body mass index (BMI) as covariates were undertaken. Four synonymous (P7P, T32T, G37G and V387V), three missense (S12C, I32T and L478P) and one nonsense (Y319X: g1230757 C > G) polymorphisms were found. One of the synonymous polymorphisms was novel (V387V) by reference to the dbSNP database. The Y319X genotype distribution of CC:CG:GG in this population showed frequencies of 0.382, 0.461 and 0.156, respectively, which followed Hardy-Weinberg equilibrium. The nonsense polymorphism had odds ratios of 0.83 (confidence interval [CI] = 0.59-1.15, p = 0.26) in males and 0.96 (CI = 0.72-1.29, p = 0.80) in females with hypertension or current medication for hypertension. For the quantitative analysis, we excluded the current medication subgroup. The nonsense allele was not a significant predictor for systolic or diastolic blood pressure. This is the first report showing that a single polymorphism in SLC6A18 is not associated with hypertension or blood pressure in Japanese. PMID- 16340171 TI - Increased serum malondialdehyde levels in chronic stage of ischemic stroke. AB - Serum lipoperoxidation products such as malondialdehyde (MDA) reflect oxidative stress. There are contradictory results addressing the levels of lipoperoxidation products in chronic phase of ischemic stroke. In the present study, we aimed to determine the serum MDA levels in stroke patients 6 months after the cerebrovascular accident. We also compared serum MDA levels in two major groups of patients with ischemic stroke resulting from small vessel and large vessel diseases, respectively. Serum MDA levels of thirty-eight patients who had ischemic stroke (19 with atherothrombotic ischemic stroke and 19 with lacunar infarction) and 30 healthy volunteers were measured. While there was no significant difference in serum MDA levels between the chronic ischemic stroke subgroups (p = 0.795), the serum MDA levels of patients with atherothrombotic ischemic stroke (p < 0.001) or with lacunar infarction (p < 0.001) were significantly higher compared to the control group. We also demonstrated that serum MDA levels of the patients with and those without hypertension (p = 0.846), diabetes mellitus (p = 0.891), or dyslipidemia (p = 0.38) were not significantly different. In conclusion, serum MDA levels were elevated in chronic stroke patients with small or large vessel diseases. To the best of our knowledge, this is the first report showing that serum MDA levels of these two groups are not significantly different. Furthermore, serum MDA levels do not differ solely by the existence or nonexistence of hypertension, diabetes mellitus or hyperlipidemia. PMID- 16340172 TI - Atopy in patients with bronchiectasis: more than coincidence. AB - Bronchiectasis is common in developing countries, but its precise underlying mechanism can be detected in only about 40% of the cases. The studies reporting the frequency of atopy and its relation to radiological findings and lung function in bronchiectasis are limited in number, and the results are controversial. The present study was designed to investigate the relationship between atopy and bronchiectasis by means of high resolution computed tomography (HRCT) and pulmonary function tests. Skin prick test, HRCT and pulmonary function tests, including spirometric values of forced expiratory volume in one second (FEV1), FEV1/FVC (forced vital capacity) ratio were performed in 121 bronchiectatic patients of unknown etiology and in 68 healthy controls. Atopy and HRCT scores for the severity of atopy and extent of bronchiectasis respectively were determined for each patient. The rate of atopy (48.8% vs 11.8%) and mean atopy score (14.3 +/- 10.1 mm vs 5.5 +/- 2.1 mm) were significantly higher in patients with bronchiectasis than those in controls. Atopic patients had significantly worse spirometric values and more extended bronchiectasis than non atopics. There is a significant correlation between atopy and HRCT scores (r = 0.54, p < 0.001), indicating that the more severe atopy is the more extended bronchiectasis. In conclusion, we suggest that the rate of atopy is higher in bronchiectatic patients than that in healthy controls. Bronchiectatic patients with atopy have lower spirometric values and higher HRCT scores. Atopy might be considered as a deteriorating and/or a causative or contributing factor for development of bronchiectasis. PMID- 16340173 TI - Lack of an association between a newly identified promoter polymorphism (-1702G > A) of the leukotriene C4 synthase gene and aspirin-intolerant asthma in a Korean population. AB - Aspirin-intolerant asthma (AIA) is a distinct clinical syndrome that refers to the development of bronchoconstriction in asthmatic individuals following the ingestion of aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs). It is widely recognized that increased cysteinyl leukotriene (cysLT) biosynthesis is associated with the development and progression of AIA. Leukotriene C4 synthase (LTC4S) is the terminal enzyme in cysLT production and is a strong candidate gene in the pathogenesis of aspirin-intolerant asthma (AIA). In this paper, we report a new single nucleotide polymorphism (SNP) of the LTC4S promoter, -1702G>A, in AIA patients and evaluate its genetic role in the association with the LTC4S-444 A>C polymorphism. We enrolled 110 AIA patients, 125 aspirin-tolerant asthma (ATA) patients, and 125 normal controls. SNP genotyping of the LTC4S-1702G>A and 444A>C polymorphisms was performed using SNP-IT assays. Haplotype analyses were performed using Haploview version 2.05, which is based on an estimation maximization (EM) algorithm. There were no significant differences in the allele or genotype frequencies of the LTC4S-1702G>A and -444A>C polymorphisms among the three groups (p > 0.05), with no significant differences in the observed haplotype frequencies (p > 0.05). Moreover, no significant associations were found between the genotype of each SNP in AIA patients with the clinical characteristics, including a forced expiratory volume in one second (FEV1) %, a provocation concentration of methacholine to induce more than 20% decrease of FEV1 (PC20) to methacholine, and serum total IgE levels (p > 0.05). These results indicate that there is no association between these two promoter polymorphisms of LTC4S and the phenotype of AIA in a Korean population. PMID- 16340174 TI - A simple quantitative assay for urinary adenosine using column-switching high performance liquid chromatography. AB - Adenosine is a physiologically active molecule produced locally in many sites of the body to regulate various cell functions. Measurement of levels of the factor in organs and biological fluids provides clues to its role and we reported an accurate quantitative high-performance liquid chromatography method for urinary adenosine requiring no preliminary sample preparation, other than filtration. Analyses were performed isocratically with a reversed-phase and a molecular exclusion columns connected by a column switch. Each sample was analyzed automatically in 35 min. Linearity could be verified up to 1,000 micromol/L (r = 0.999) and recovery of adenosine was 94.6-98.0%. The coefficients of variation (CV) were established to be 0.56-1.32%, intra-assay, and 1.61-4.67%, inter-assay. Based on analyses of healthy individuals at different ages, we are here able to provide age-related values, infants (1.51 +/- 0.71 micromol/mmol creatinine) and children (1.06 +/- 0.36 and 0.83 +/- 0.27 micromol/mmol creatinine; aged 1-5 and 6-10 years), excreting significantly higher amounts of adenosine than adults (0.44 +/- 0.08 micromol/mmol creatinine). We also measured urinary adenosine from patients suffering from metabolic disease or severe respiratory failure and found that unfavorable pathophysiologic conditions are associated with appreciable elevation of adenosine. PMID- 16340175 TI - Magnetic resonance imaging of morphological and functional changes of the uterus induced by sacral surface electrical stimulation. AB - The purpose of this study is to examine the morphological and kinematical changes of the uterus induced by electrical stimulation applied to the skin just above the second and fourth posterior sacral foramens (sacral surface electrical stimulation [ssES]) in 26 healthy subjects. Out of them, eight subjects who had severe pain subjectively during every menstruation received ssES just in menstruation. Morphological and functional changes of the uterus were examined by using T2-weighted magnetic resonance (MR) imaging and T1-weighted MR cinematography, respectively. Cyclic electrical stimulation for 15 min with 5 sec ON and 5 sec OFF was applied just before MR scanning. A decrease in thickness of the muscular layer of the uterus was observed in every subject after ssES for 15 min and was significant as compared with the thickness before ssES. Periodic uterine movement during menstruation was observed in the subjects with severe menstrual pain in MR cine and the power spectrum analysis of the movement showed a marked decrease in peak power and frequency after ssES treatment. We conclude that ssES causes a reduction of static muscle tension of the uterus in all menstrual cycle periods and suppression of uterine peristalsis during menstruation in the subjects with severe menstrual pain. Possible neural mechanisms for these static and dynamic effects of ssES on the uterus at spinal level are discussed. PMID- 16340176 TI - Increased plasma levels of growth hormone, insulin-like growth factor (IGF)-I and IGF-binding protein 3 in pregnant rats with exercise. AB - Growth hormone (GH) and insulin-like growth factor-I (IGF-I) are closely related molecules. Insulin-like growth factor-binding protein-3 (IGFBP-3) is a main molecule that binds IGF-I. GH, IGF-I and IGFBP-3 have important roles in growth and development. In this study, we investigated the effects of exercise during pregnancy on maternal plasma levels of GH, IGF-I and IGFBP-3 and on fetal development. We also recorded the weights of placenta, lengths of umbilical cord, fetal body weights, fetal heights, and weights of fetal tissues. Pregnant Wistar Albino rats were divided into two groups: exercise and control groups (n = 7 for each). A treadmill exercise was performed as 20 m/min for 20 min/day, once per day for 19 days in exercise group. Blood samples were collected from pregnant rats on 0, 7th, 14th and 20th days of gestation (D) under anesthesia with intracardiac puncture, and maternal plasma levels of GH, IGF-I and IGFBP-3 were determined. Fetuses were taken with cesarean section on D20, and various parameters for fetal growth were measured. Plasma GH and IGF-I levels were elevated in exercising pregnant rats on D14 and D20, respectively, when compared to controls, and IGFBP-3 levels were increased on D14 and D20. Among the growth parameters examined, only fetal body weights and weights of fetal liver were significantly decreased in the exercise group (p < 0.01 and p < 0.05, respectively). These results indicate that maternal exercise significantly increases plasma levels of GH, IGF-I and IGFBP-3 in the late period of pregnancy but causes adverse effects on fetal growth. PMID- 16340177 TI - New body mass index criteria of central obesity for male Japanese. AB - In recent years, intra-abdominal visceral fat leads to obesity-related complications. A simple indicator that reflects the mass of visceral fat is also needed to enable practical screening of patients. The present study was designed to establish new body mass index (BMI) criteria of central obesity for male Japanese. The subjects were 516 men aged from 19 to 80 years old who were examined at the physical examination center in the regular health check conducted by their company. Correlations between visceral fat area (VFA) or subcutaneous fat area (SFA) and BMI in the subjects were investigated. Receiver Operating Characteristics (ROC) curve was used to find out the optimal cut-off values of BMI to predict central obesity. We compared the percentile ranks corresponding to VFA of 100 cm2, BMI of 25 kg/m2 and new BMI criteria to check to see whether the present BMI criteria classify correctly Japanese men as central obesity. Further evidence for the effectiveness of BMI for VFA is needed. The correlation coefficient between VFA or SFA and BMI was 0.59 or 0.67, respectively. At the cut off for BMI that maximized sensitivity and specificity for predicting central obesity was 24 kg/m2. Moreover, the percentile value corresponding to VFA of 100 cm2, BMI of 24 kg/m2, and BMI of 25 kg/m2 was the 53, 50 and 61 percentile, respectively. It is necessary to lower a cut-off point for central obesity from BMI of 25 kg/m2 to 24 kg/m2. PMID- 16340178 TI - Prevalence and clinical characteristics of migraine in university students in Turkey. AB - Migraine is a significant health problem due to its frequency and accompanying morbidity, which includes disability and loss of performance. We aimed to estimate the prevalence of migraine and assess the disability and preferences of treatment among university students in Afyon, a Turkish city. This is the first cross-sectional prevalence study of migraine conducted on university students in Turkey. The study has been carried out in two stages. The first stage aimed to identify the students with migraine by using a standardized International Headache Society (IHS) questionnaire. In this questionnaire, the students were asked about medical consultations and medicines used during attacks. The standard questionnaire was applied to 1,029 students, and migraine was found in 128 students (12.4%). Of these headache sufferers, 104 students were using non prescribed drugs during headache attacks, while the remaining 24 students were using the drugs on advice of physicians. At the second stage, we evaluated the impact of disease on daily life in these students by the Migraine Disability Assessment Scale (MIDAS) questionnaire, showing that the degrees of disability are minimal in 11 students (8.6%), mild in 30 students (23.4%), moderate in 34 students (26.6%), and severe in 53 students (41.4%). In conclusion, migraine attacks are associated with a considerable degree of handicap in activities of daily living. However, many university students with migraine do not consult a physician and continue to treat their headaches with simple analgesics. Global educational programs are required for patients to recognize the importance of effective migraine treatment. PMID- 16340179 TI - Concerning the article by T. Curstedt and J. Johansson: new synthetic surfactants - basic science. PMID- 16340180 TI - Surfactant therapy: present and future perspectives. Proceedings of the 21st International Workshop on Surfactant Replacement. June 1-4, 2006. Oslo, Norway. PMID- 16340181 TI - Urinary S-100B concentrations in term and preterm infants at risk for brain damage. AB - BACKGROUND: Elevated serum concentrations of S-100B, a 21-kDa protein expressed in astroglial cells, has been used to assess cerebral damage after head trauma, infection, ischemia, and perinatal asphyxia. OBJECTIVE: As S-100B is eliminated by the kidneys, we investigated the feasibility of measuring S-100B in urine of newborns with severe perinatal asphyxia, and in very low birth weight (VLBW) preterm infants at risk for neurodevelopmental impairment. METHODS: We first analyzed urine samples of 8 term or near-term newborns without major medical problems, followed by urine samples of 2 term newborns with severe birth asphyxia, and finally urine samples of 8 VLBW (gestational age 24-28 weeks) infants collected every 4 h for up to 10 days. RESULTS: Urinary S-100B concentrations in 8 term or near-term newborns without major medical problems were consistently <1 microg/l. In 2 term newborns with severe asphyxia (Apgar 0/0/0 and 0/2/4) who subsequently had widespread cerebral damage on magnetic resonance imaging, peak urinary S-100B concentrations on the first day of life were 28.1 and 28.4 microg/l, respectively. In 5/8 VLBW infants, urinary S-100B was> microg/l in samples obtained on the first day of life (range 1.2-44.9 microg/l, median 6.8 microg/l). Peak S-100B in urine samples collected during the first 12 h of life were negatively related to gestational age (R(s)=-0.882, p=0.009). Three of the 8 preterm infants had peak urinary concentrations>0 microg/l but neither ultrasound signs of brain damage nor neurodevelopmental delay at 1 year corrected age. CONCLUSIONS: The determination of urinary S-100B concentrations might be helpful in term infants with severe asphyxia, while high urinary S-100B concentrations in preterm infants are to be attributed to immaturity. PMID- 16340182 TI - On-line filtration therapies: emerging horizons. PMID- 16340183 TI - The frequency and determinants of calcification in intracranial arteries in Chinese patients who underwent computed tomography examinations. AB - BACKGROUND: Intracranial artery calcification is common but the prevalence and determinants are not well established. We aim to describe the prevalence and location of calcification in intracranial arteries according to brain multi detector-row computed tomography (MDCT) images, and to investigate its correlation with potential risk factors. METHODS: We studied consecutive men and women referred for brain CT in December 2004. All patients received a questionnaire regarding their medical history related to atherosclerosis, including traditional risk factors of atherosclerosis, serum chemistry values and inflammatory markers. All CT examinations were done with a 16-slice MDCT and the severity of intracranial artery calcification was categorized. RESULTS: Four hundred and ninety patients aged 1.4-101 years (62.92+/-19.04; mean+/-SD) were included in our study. There were 340 patients (69.4%) who had intracranial artery calcification. The highest prevalence of intracranial artery calcification was seen in the internal carotid artery (60%), followed by vertebral artery (20%), middle cerebral artery (5%) and basilar artery (5%). Patients with calcification were significantly older than those without calcification (p<0.001). A significantly higher prevalence of calcification was present among patients with hypertension (p<0.001), diabetes (p<0.001), renal failure (p<0.05), atrial fibrillation (p<0.05), smoking (p<0.05), hyperlipidemia (p<0.001), ischemic heart disease (p<0.05) and ischemic stroke (p<0.001). Mean values of serum phosphate, serum urea and CRP level were also significantly higher in patients with intracranial artery calcification (p<0.05, respectively), and there was a trend that patients with intracranial calcification had a higher white blood cell count (p=0.070). Stepwise multiple logistic regression showed age (RR=2.795 per 10 years), a history of ischemic stroke (RR=3.915), and white blood cell count (RR=1.107) to be independently associated with intracranial artery calcification. CONCLUSIONS: Calcification of the intracranial arteries is associated with age, history of ischemic stroke and white blood cell count. Further prospective studies to investigate the clinical significance of intracranial artery calcification are needed. PMID- 16340184 TI - Dipyridamole decreases protease-activated receptor and annexin-v binding on platelets of post stroke patients with aspirin nonresponsiveness. AB - BACKGROUND: Although controversial, the phenomenon of aspirin resistance (AR) has been correlated in some small studies with poor clinical outcomes in patients with coronary artery disease. Even less is known regarding the role of AR in the post stroke population. The reason for and the underlying mechanism of AR is unknown. We hypothesized that excessive formation of thrombin on the platelet surface may contribute to this phenomenon and assessed how dipyridamole affects multiple platelet and thrombin generation biomarkers in AR patients after ischemic stroke. METHODS: Whole blood samples from 20 post stroke AR patients were pretreated with dipyridamole, simulating the therapeutic range, and then incubated for 45 min at 37 degrees C. Platelet characteristics were assessed by aggregometry, cartridge-based analyzer, and receptor expression by flow cytometry. Markers of thrombin generation were measured in the autologous plasma by ELISA. RESULTS: Pretreatment of blood with dipyridamole resulted in 22-26% diminished expression of intact PAR-1 receptor (p=0.021 and p=0.024) and 28-31% decrease of annexin V binding (p=0.031 and p=0.02) after incubation with 2 microg/ml and 4 microg/ml of dipyridamole, respectively. Platelet aggregation and thrombin generation markers were not affected in vitro by dipyridamole. CONCLUSIONS: Dipyridamole may be capable of overcoming increased prothrombinase complex formation and be in part able to compensate for AR in patients with moderate carotid stenosis. This phenomenon may explain the clinical advantages of Aggrenox, known to reduce ischemic events in post stroke patients as proven in clinical trials, though an additional antithrombotic benefit beyond the platelet inhibition by aspirin alone. PMID- 16340185 TI - Effect of the Glycine Antagonist Gavestinel on cerebral infarcts in acute stroke patients, a randomized placebo-controlled trial: The GAIN MRI Substudy. AB - BACKGROUND AND PURPOSE: Gavestinel, GV150526, is a selective antagonist at the glycine site of the N-methyl-D-aspartate receptor. The safety and efficacy of GV150526 were studied in two phase III randomized placebo-controlled clinical trials of acute ischemic stroke patients within 6 h from onset [The Glycine Antagonist in Neuroprotection (GAIN) International and GAIN Americas Trials]. A planned MRI substudy within these trials investigated the effect of gavestinel on infarct volume. METHODS: Patients enrolled in the GAIN trials at designated MRI substudy sites were eligible if they had a pretreatment acute cortical lesion on diffusion-weighted MRI of at least 1.5 cm diameter or 5 cm(3). Final lesion assessment was performed on T(2)-weighted MRI at month 3. Blinded image analysis was performed centrally. The primary hypothesis was that gavestinel would attenuate lesion growth from baseline relative to placebo. RESULTS: A total of 106 patients were eligible, 75 (34 gavestinel, 41 placebo) of whom had month 3 scans (primary analysis population). No effects of gavestinel on infarct volume were observed in the primary or other analyses. However, significant associations of lesion volume to clinical severity and outcomes were observed. Ischemic lesion volume decrease was predictive of substantial clinical improvement. CONCLUSION: Consistent with the clinical outcomes in the GAIN trials, no effects of gavestinel on ischemic infarction was observed. Concordance of results of the clinical outcome trials with those of this infarct volume substudy as well the associations of infarct volume to clinical outcomes further support the potential role of infarct volume as a marker of outcome in dose finding and proof of principle acute stroke trials. PMID- 16340186 TI - Correlation of coronary and cerebral atherosclerosis: difference between extracranial and intracranial arteries. AB - BACKGROUND: A difference with regard to the correlation with coronary atherosclerosis (CAS) between extracranial carotid atherosclerosis (ECAS) and intracranial cerebral atherosclerosis (ICAS) has been assumed but not proven clearly by direct comparison within the same population. METHODS: A consecutive series of 246 patients undergoing coronary artery bypass graft surgery were reviewed. The severity of CAS was estimated as a CAS score based on coronary angiography. The presence of ECAS and ICAS was screened by transcranial Doppler and carotid duplex sonography, and confirmed by magnetic resonance angiography. RESULTS: The CAS scores in patients with ECAS were observed to be higher than those in patients without ECAS (10.62+/- 4.80 vs. 9.45+/- 4.25; p=0.054 on the Mann-Whitney U test). The difference in CAS scores was smaller between patients with and without ICAS (10.41+/- 4.44 vs. 9.66+/- 4.49; p=0.201). Similar patterns were observed on comparing the correlation of ECAS and ICAS with a quartile of the CAS score. An advanced CAS, which was generated by collapsing the quartiles of the CAS score into 75th percentile or less and more than the 75th percentile, was significantly associated with ECAS, but not with ICAS. These associations remained unchanged after adjustments had been made for age, sex, hypertension, diabetes mellitus, hyperlipidemia, smoking and a history of stroke or transient ischemic attack. CONCLUSIONS: This study suggests that the correlation of CAS with ECAS is stronger than that of CAS with ICAS, and this difference is independent of the classic risk factors for atherosclerosis. PMID- 16340187 TI - Diazepam to improve acute stroke outcome: results of the early GABA-Ergic activation study in stroke trial. a randomized double-blind placebo-controlled trial. AB - BACKGROUND: We tested whether diazepam, a GABA-ergic drug that also inhibits brain nitric monoxide formation, improves acute stroke prognosis. METHODS: 880 patients, randomized within 12 h of acute stroke, received diazepam 10 mg or placebo by rectiole, as soon as possible, followed by 10-mg tablets twice daily for 3 days. Primary outcome was independence (Rankin score <3) at 3 months; secondary outcome was complete recovery (Barthel index >or=95 or Rankin score or =1.5 g/24 h), mesangial proliferation and elevated serum creatinine (> or =2 mg/dl) were statistically independent risk factors for renal prognosis (RR = 1.90, 2.72, 2.84, respectively). Conversely, strict blood pressure control had a favorable effect on renal prognosis. CONCLUSION: The clinico-pathological features and outcomes of IgANMHT patients were different from those of PMHT patients. The renal survival of IgANMHT patients was poor, which was determined by many factors. Early control of proteinuria, early monitoring and strictly controlling blood pressure may contribute to the renal survival. PMID- 16340216 TI - Hyperlipemia and oxidation of LDL induce vascular smooth muscle cell growth: an effect mediated by the HLH factor Id3. AB - Hyperlipemia and oxidized LDL (ox-LDL) are important independent cardiovascular risk factors. Ox-LDL has been shown to stimulate vascular smooth muscle cell (VSMC) proliferation. However, the effects of hyperlipemia and the molecular mechanisms mediating hyperlipemia and ox-LDL effects on VSMC growth are poorly understood. The helix-loop-helix (HLH) transcription factor, Id3, is a redox sensitive gene expressed in VSMC in response to mitogen stimulation and vascular injury. Accordingly, we hypothesize that Id3 is an important mediator of ox-LDL and hyperlipemia-induced VSMC growth. Aortas harvested from hyperlipemic pigs demonstrated significantly more Id3 than normolipemic controls. Primary VSMC were stimulated with ox-LDL, native LDL, sera from hyperlipemic pigs, or normolipemic pigs. VSMC exposed to hyperlipemic sera demonstrated increased Id3 expression, VSMC growth and S-phase entry and decreased p21cip1 expression and transcription. Cells stimulated with ox-LDL demonstrated similar findings of increased growth and Id3 expression and decreased p21cip1 expression. Moreover, the effects of ox LDL on growth were abolished in cells devoid of the Id3 gene. Results provide evidence that the HLH factor Id3 mediates the mitogenic effect of hyperlipemic sera and ox-LDL in VSMC via inhibition of p21cip1 expression, subsequently increasing DNA synthesis and proliferation. PMID- 16340219 TI - A centenary of auscultatory blood pressure measurement: a tribute to Nikolai Korotkoff. AB - In 1905, Dr. Nikolai Korotkoff (1874-1920), a Russian surgeon, discovered a simple and precise technique to measure arterial pressure. He was born on 26th February 1874 in the central Russian city of Kursk. Korotkoff graduated from the Medical Faculty of Moscow University in 1898, but he worked later in the Surgical Clinic at the Imperial Military Medical Academy in St. Petersburg (Russia). Korotkoff served as a military surgeon during the Russo-Japanese War (1904-1905) and his major efforts were to find reliable clinical signs that could predict whether limb flow would be viable after vascular surgery of traumatic aneurysm. He found that after complete compression, the aneurysm of the arm (i.e. distal pulse on a. brachialis) disappeared with Riva-Rocci cuff and then gradually decreasing the pressure, a series of sounds could be heard by stethoscope under the artery distal to the compression. Korotkoff described four distinct phases of sounds: first sound, then compression murmurs, second tone, and disappearance of sounds. Korotkoff was also able to demonstrate the same auscultatory finding in healthy persons. He failed to notice only the muffled second sound, which was demonstrated a little later. These classical observations are now well known as the five different phases of Korotkoff sounds. In November 1905, during a conference of the Imperial Military Medical Academy, he reported his discovery in a short presentation entitled 'On the issue of the methods for measuring blood pressure'. In 1939, the Joint Committee of the American Heart Association and the Cardiac Society of Great Britain and Ireland recognized officially and accepted worldwide Korotkoff's method for blood pressure determining. PMID- 16340221 TI - Common pitfalls in the conduct of clinical research. AB - Recently, paradigm shift from expert opinion towards evidence-based medicine has occurred encouraging physicians to base their treatment decisions on the best available research evidence. In the hierarchy of evidence randomized clinical trials (level 1 evidence) are considered of the highest quality (least biased) while non-randomized studies represent lower levels (levels 2-4). Several pitfalls in the design and conduct of clinical research include: lack of randomization, lack of concealment, lack of blinding, and errors in hypothesis testing (type I and II errors). A basic understanding of these principles of research will empower both investigators and readers when applying the results of research to clinical practice. PMID- 16340222 TI - A journey across the wall of varicose veins: what physicians do not often see with the naked eye. AB - OBJECTIVE: To examine varicose veins (VVs) from inside out in order to help surgeons and general practitioners better understand the pathogenesis of the disease and improve their management. MATERIAL AND METHODS: A comprehensive examination of the wall of VVs was performed using transmission electron microscopy. The ultrastructural morphology of the collagen, elastin and smooth muscle content of the wall was analyzed in a sample of 10 patients (4 male and 6 female) and 10 matched controls aged between 37 and 50 years. RESULTS: Analysis of the tunica media revealed that the smooth muscle cells were significantly separated from each other by a marked increase in amorphous and fibrous tissue in which many of the collagen and elastin fibers lost their normal structural arrangement. The cells contained a large number of membrane-bound intracellular vesicles and cytoplasmic vacuoles. The collagen fibers were smaller and thinner than what is commonly seen in normal veins, and they were widely separated from each other. A light electron-lucent center was observed in the middle of the fibers. Similar changes were also seen in the intima and were associated with irregular plaque-like intimal thickening. CONCLUSION: Our study revealed a significant separation among smooth muscle cells in the wall of VVs, and the presence of an abnormal amorphous extracellular matrix and intracytoplasmic vacuoles could reflect 'unusual' possible secretory and phagocytic roles of smooth muscle cells. This could provide an important explanation for the abnormal contractile function of these cells in VVs. PMID- 16340223 TI - Caries prevalence and treatment needs of 12-year-old children in the Islamic Republic of Iran. AB - OBJECTIVE: To assess the prevalence of dental caries and treatment needs of 12 year-old children living in Teheran and a rural area surrounding Esfahan. SUBJECTS AND METHODS: A total of 1,102 pupils were examined clinically for caries according to WHO criteria. One calibrated dentist examined all children. RESULTS: Of the children studied, 63.8% had sound permanent dentition, 9.9% had filled teeth and 26.3% needed treatment. The mean DMFT value found was 0.77 (DFS: 1.19). The D component was 58.7%, the M component 3.4% and the F component 37.9%. The levels of caries experience varied according to the child's residence. The teeth of children living in the rural area (mean DMFT: 0.38) were significantly less affected than those of children living in Teheran (mean DMFT: 1.1). In the capital city of Teheran, considerable differences related to social status were observed. The lowest DMFT values (0.74) were found in southern Teheran where the poorest people live. The highest caries experience was found in the middle-class social stratum. CONCLUSION: The results indicate that caries prevalence among 12 year-old children in the Islamic Republic of Iran is as low as in the developed countries of central Europe. PMID- 16340224 TI - Clinical effect of recalibration of the roche cardiac troponin T assay. AB - OBJECTIVE: To assess the effect of recalibration of the Roche 3rd-generation cTnT assay with recombinant human cardiac troponin T (cTnT) standards on clinical decision limits. MATERIALS AND METHODS: Serum samples from 77 patients (66 +/- 16 years) admitted to the coronary care unit were assayed using the 2nd- and 3rd generation cTnT assays. RESULTS: There was excellent agreement (r = 0.99 Spearman, 95% CI 0.99-1.0; p or =1 cm) predicted both overall and disease-free survival (p < 0.050). Histology (serous vs. others) (p = 0.023) also influenced overall survival. Overall 5-year survival was 38% when serum hCGbeta was below 3.5 pmol/l, while it was 18% when the level was higher (p = 0.052). The corresponding disease-free 5-year survival was 38 and 20%, respectively (p = 0.014). Patients with CA 125 values above 1,017 kU/l had an overall 5-year survival of 39% as compared with 14% for those with lower values (p = 0.009), while the disease-free survival was 37 and 23%, respectively (p = 0.096). Serum TATI was not a prognostic marker. Serum concentrations of hCGbeta and CA 125 correlated significantly with stage (p = 0.049 and p = 0.050, respectively). In multivariate Cox proportional hazards regression analysis, only hCGbeta, stage and histology emerged as independent prognostic factors. CONCLUSIONS: Clearly elevated serum concentrations of hCGbeta and CA 125 predict survival in fallopian tube carcinoma, but in multivariate analyses, only hCGbeta is a prognostic factor independent of stage and histology. PMID- 16340250 TI - Characterization of integrin and anchorage dependence in mammary epithelial cells following c-erbB2-induced epithelial-mesenchymal transition. AB - Signalling from the proto-oncogene c-erbB2 in mammary epithelial cells has earlier been shown to result in epithelial-mesenchymal transition (EMT) giving rise to fibroblast-like cells, and acquisition of anchorage-independent growth (AIG) usually determined by growth capacity in soft agar. In this study, we have analysed AIG associated with c-erbB2-induced EMT in a human mammary epithelial cell line. Intriguingly, cells capable of growth in soft agar were shown to be dependent on the function of beta(1) integrin extracellular matrix receptors for growth in collagen. We therefore tested the hypothesis that apparent AIG was due to deposition of extracellular matrix in the agar. Although the fibroblastic cells had strongly upregulated expression of the fibronectin receptor subunit integrin alpha(5) andabundant fibronectin fibrils, these properties did not have a positive correlation with AIG. Furthermore, antibody blocking of integrin alpha(5) and beta(1) failed to inhibit AIG. These results indicate that the anchorage-independent cells are not dependent on connection to extracellular matrix, but instead may be subject to a growth-inhibitory effect from the collagen in the absence of integrin signalling. This notion was supported by the finding that integrin blocking of the fibroblastic cells in fibrin was without effect on proliferation. PMID- 16340252 TI - Quo Vadis--post graduate medical education in Nepal? PMID- 16340253 TI - Study of patients presenting with symptoms of peripheral neuropathy and thickened greater auricular nerve. AB - OBJECTIVE: To analyze symptoms and make a clinical diagnosis of leprosy in patients presenting with symptoms of peripheral neuropathy and found to have thickened greater auricular nerve. DESIGN: Cross-sectional study. SETTING: Kathmandu Medical College Teaching Hospital, Sinamangal, Kathmandu. Patients attending the medical out-patient department of this hospital were taken in the study. MATERIALS AND METHODS: 40 patients presenting with symptoms of peripheral neuropathy and are found to have thickened greater auricular nerve were included. RESULTS: Thickened greater auricular nerve and sensory symptoms showed male predominance (97.5%). Mean age of involvement was 28.65 years. The symptom most frequently complained of was chest pain (75%), followed by pins and needle sensation 67.5%, burning of the upper extremities, nape of the neck and chest 57.5%, palpitation 45%, disturbed sleep mostly said to be due to burning 35%, sweating 20%, dizziness 17.5%, shortness of breath 7.5%, and numbness of the limbs in 2.5%. None of the patients had somatic neuropathy. Autonomic neuropathy was present in 42.5 %. CONCLUSIONS: 1. In endemic areas patients with thickened peripheral nerve and sensory symptoms should be diagnosed clinically as primary neuritic leprosy. 2. In absence of objective loss of somatic sensation autonomic neuropathy may be the only early indicator of neuritis. 3. Close follow up of these patients is necessary. PMID- 16340254 TI - Comparison of Pap test among high and non-high risk female. AB - A prospective study of pap smear in 100 high risk and equal number of non high risk female among total 1022 female Gynecological patients within a period of two and half months {Beginning of Sept. to middle of Nov. 1995} in Maternity Hospital, Thapathali is presented. There were 9 cases positive for dyskaryosis among high risk and 3 cases among the comparison group. All positive cases were at the age of 35 years and above. In 9 positive cases, 5 cases were in CIN I (55.55%) while 4 were in CIN II (44.44%). Similarly out of 3 positive cases in comparison group 1 was in CIN I category (33.33%) and 2 cases were in CIN II (66.66%). Relation of positive cases with low social class revealed 80% CIN I and 50% CIN II among high risk group where as 66.6% CIN II in comparison group. Analysis of risk factor in development of various grades Dyskaryosis revealed 60% of CIN I had high parity while 50% had CIN II. There are about 40% of CIN I and 75% CIN II among child birth less than 19yrs, 60% smoker had CIN I where as 100%. Smoker had CIN II. 80% of CIN I gave history of excessive vaginal discharge where as 50% of CIN II had excessive vaginal discharge. 40% of CIN I was having injection Depo provera. Cases were also analyzed according to risk factor. Out of 9 positive cases among high-risk females 5 positive had parity more than 4 and 4 cases had less than 4. 5 positive cases were among less than 19 years of first childbirth, 4 among more than 19 years. 7 positive cases were smoker and 2 positive cases were non-smoker. 6 positive cases gave history of abnormal vaginal discharge and 3 positive cases had no abnormal vaginal discharge. Out of 9 positive cases 2 had history of injection Depo provera continuously for more than 5 years where as 7 were non users. PMID- 16340255 TI - Epilepsy in children: an epidemiological study at Kathmandu Medical College Teaching Hospital Kathmandu. AB - OBJECTIVES: To analyze the relative frequencies of various epileptic seizures and to study the age at onset of different seizure types in Nepalese children. DESIGN: Prospective study. SETTING: Hospital outpatient based in Kathmandu, Nepal, between November 2001 to October 2002. PARTICIPANTS: 50 children diagnosed as epilepsy excluding neonatal and febrile seizures. MAIN OUTCOME MEASURE: Diagnosis and classification of cases according to the International Classification of Epilepsy of the International League Against Epilepsy [ILAE] and number of patients in each category with various ages at first seizure. RESULT: Generalized seizures (78%) were 3.54 times commoner than partial seizures (22%). Most frequent seizure types were generalized tonic clonic (36%), tonic (16%), complex partial (14%), atonic (12%) and absence (10%). Generalized clonic, simple partial and partial with secondary generalization, each had less than 5% frequencies. In 40% cases the first seizure occurred when aged between 2-5 years. In partial seizures the peak age at onset was observed below 6 years while primary generalized seizure was more frequently seen in age group 2-10 years. CONCLUSION: More paediatric patients with primary generalized seizures (78%) were observed than with partial seizures (22%). In this age group, the most frequent seizure type was generalized tonic clonic (36%) with the peak frequency of age at onset of seizures in 2-5 years. PMID- 16340256 TI - A profile of menstrual disorders in a private set up. AB - Menstruation and its disorders are still considered unholy and impure and are not yet recognised as significant reproductive health morbidity. Therefore a prospective study was carried out at a private clinic for a period of three months where total number of patients coming with current or past menstrual problems are 525. This number did not include pregnant women or those on any hormonal medications or having dysfunctional uterine bleeding. This study aimed to find out the incidence of Menstrual Morbidity and their mode of presentation. It has also tried to find out these women's age, parity, age of menarche and number, following discriminating traditional rituals during their 1st and regular menses, their family planning status and the districts from where they came to Kathmandu for their treatment. In this study, menstrual morbidity was found to be 43.75%. Approximately 60% of women having menstrual complaints had absolutely normal menstrual cycle; whereas 13% of them had irregular, 17% of them had prolonged and 6% had short menstrual cycle. A significant number (46%) of women although suffering from menstrual problems presented with other symptoms like vaginal discharge, pain lower abdomen, subfertility, urinary problems, abdominal lumps and for cuT check-ups. 3% of the women who presented with vague, non specific complaints asking for a general check up had one or the other menstrual problem. Although approximately 69% of these women were from the age group of 20 39 years, 4% of them were adolescents and 27% above 40 years. It was observed that although approximately 78% of these women were primi- and multiparous ladies, 22% were unmarried and nulliparous suffering from various menstrual morbidities. More than 55% of these women had their menarche at the age of 12-14 years. It was not surprising to note that more than 90% of women had to follow the traditional unhealthy and unsociable rituals during their first menstruation. More than 75% of them had to follow the discriminating traditional rituals which consider a menstruating woman untouchable for 5 days of every month throughout their active reproductive lives. 20% of these women were using non-hormonal contraceptive methods, out of which more than 50% had undergone permanent sterilisation. It was a matter of pride to note that this clinic was providing the health care services to the adolescents and women of 13 zones and more than 52 districts of the country. PMID- 16340257 TI - Incidence of ophthalmoscopic fundus changes in hypertensive patients. AB - A prospective, hospital based, clinical study was conducted in hypertensive patients referred to the eye OPD, KMCTH, Sinamangal during a period of three months to find out the incidence of fundus changes. A total of 302 hypertensive patients were included in the study and their fundus changes were evaluated by direct ophthalmoscopy. The age of the patients ranged from 30-70 years and the duration of hypertension from 1-25 years. The blood pressure was not controlled in 218 patients (72.18%). More female patients (56.22%) were hypertensive than male (43.78%). But the fundus changes were less in female patients. Caste-wise hypertension was more common in Brahmins (38.41%) and fundus changes were also comparatively more. The fundus changes were found in 192 patients (63.57%) and the most common findings were hypertensive retinopathy. GrI + GrII combined together (52.31%). The other common fundus finding was BRVO in 11 patients (3.64%). A routine ophthalmoscopic fundus examination to detect the retinal changes in hypertensive patients is recommended. PMID- 16340258 TI - Study of pterion in skulls of Awadh area--in and around Lucknow. AB - Two hundred and three skulls of known sex (100 male and 103 female) were studied to find out the variations of Pterion. Sphenoparietal variety of Pterion was seen predominantly along with frontotemporal and stellate varieties. PMID- 16340259 TI - Road traffic accident (RTA) as major killer: a report on medico-legal autopsies in Bharatpur hospital. AB - Road traffic accidents are the major cause of medico-legal deaths. In present study out of 229 autopsy cases in one year, 110 deaths were due to road traffic accident (R.T.A.) which shows the magnitude of the problem in Chitwan and adjacent districts in Nepal. PMID- 16340260 TI - A typical case of pellagra. PMID- 16340261 TI - A case report of Wilson's disease. AB - Wilson's disease (hepatolenticuler degeneration), an inborn error of copper metabolism, is an autosomal recessive disorder characterized by degenerative changes in brain, liver disease and Kayser Fleisher (KF) rings in the cornea. It is due to a defect of p-type ATPase which is probably required for normal extrusion of copper from cells. In this case report, we present a seven and half year old male who presented with complaints of slurring of speech, drooling of saliva, intentional tremor and dark pigmentation over face and trunk for last 9 months. On examination KF ring was present, spleen was palpable and intentional tremor was present. Laboratory investigations confirmed the diagnosis. PMID- 16340262 TI - Malignant teratoid medulloepithelioma in eye. AB - Malignant teratoid medulloepithelioma is an extremely rare tumor occurring in children younger than 5 years of age, arising from ciliary body epithelium or iris but few arise from optic nerve and retina. This report concerns a 5 years old boy who presented with pain, redness and protrusion of right eye. Histopathologically, the tumor was composed of epithelial and sarcomatoid component. The pseudostratified primitive appearing epithelial cells were arranged mainly in diffuse pattern, nests cords and tubules. At places, pseudo rosette and true rosette were seen. Mitoses were frequent consisting of 7-10/ HPF. The sarcomatoid component consisting of spindle shaped cells arranged in interlacing bundle were also seen. Mitoses counted 5-7/HPF. Massive areas of necrosis and hemorrhage along with calcification, focal area of mature cartilage were present. Vascular and optic nerve invasions were seen. This case of malignant teratoid medulloepithelioma is the second case diagnosed in TU Teaching Hospital within the period of 10 years and reported because of its rarity. The differentiations from other tumors of the orbit such as small cell tumor were discussed. PMID- 16340263 TI - An incidence of a complete separation of the lower lobe of right lung from the rest of upper part, as observed in a cadaver in the anatomy dissection hall. PMID- 16340264 TI - Interpleural catheter technique for perioperative pain management. AB - Management of pain in surgical patient is very crucial. It is more so in thoracic and upper abdominal surgery. Lots of technique and drugs have been used to control postoperative pain including thoracic epidural analgesia. We describe a case in whom Intraoperative and Postoperative pain was managed by injecting 0.5% bupivacaine 20 ml in the interpleural space through the catheter, followed by continuous infusion of 0.1% bupivacaine 10 ml/ hour for 24 hours. The whole perioperative and postoperative period was uneventful. The first series of patients in whom this technique was used was described by Murphy in 1983, (1) who used it in patients with multiple fractured ribs and in postoperative patients after gallbladder and kidney surgery. One year later, Reiestad and Kvalheim published their results of continuous intercostal nerve block for postoperative pain relief and presented their modification of the technique, which is now termed interpleural analgesia. PMID- 16340265 TI - Neurocycticercosis--a review. PMID- 16340266 TI - Role of Nepal medical council in MBBS curriculum. AB - The development of the Bachelor of Medicine and Bachelor of Surgery (MBBS) course in Nepal is recounted. From the stage of having a single medical college to the stage of having ten is documented. The role that the Nepal Medical Council (NMC) has played in the development of medical education in Nepal has been adequately described. Comparison of the MBBS course that the three universities are running the MBBS is done and suggestions are made for possible future directions. PMID- 16340267 TI - Atopic eczema. PMID- 16340268 TI - Muscle relaxant in 21st century. PMID- 16340269 TI - Jaw tracking devices--historical review of methods development. Part I. AB - Chewing or mastication is one of the main functions of the stomatognathic system. The use of devices for quantitatively measuring mandibular motion has recently become more common in scientific and clinical use. Often, the goal has been to provide an objective basis for diagnosing musculoskeletal disorders of the jaws, to monitor the progress of active treatment methods or to evaluate prosthodontic treatment functional results. To better understand differences between various systems to record mandibular motion a review of recording methods presented over the years was made. To give fundamental description for development of existing methods review was divided in three parts. Part I includes analyses of methods using mechanical devices, photographic methods and roentgenographic methods, describing not only technologies by themselves, but also analyzing essential limitations, possible direction of the functional improvement and, specially, their scientific and clinical significance. PMID- 16340270 TI - Jaw tracking devices--historical review of methods development. Part II. AB - Chewing or mastication is one of the main functions of the stomatognathic system. The use of devices for quantitatively measuring mandibular motion has recently become more common in scientific and clinical use. Often, the goal has been to provide an objective basis for diagnosing musculoskeletal disorders of the jaws, to monitor the progress of active treatment methods or to evaluate prosthodontic treatment functional results. To better understand differences between various systems to record mandibular motion a review of recording methods presented over the years was made. To give fundamental description for development of existing methods review was divided in three parts. Part II includes analyses of electronic and telemetric methods, magnetometry and opto-electronic methods, describing not only technologies by themselves, but also analyzing essential limitations, possible direction of the functional improvement and, specially, their scientific and clinical significance. PMID- 16340271 TI - The masticatory system--an overview. AB - Masticatory muscle physiology has been evaluated mostly from electromyographic recordings. However, electromyography coupled with jaw-tracking devices has provided much more information of the correlation between jaw movements and muscle activity. Knowledge of how the mandible moves during mastication has greatly influenced procedures in clinical dentistry. The aim of this overview is to give basic description of the classical studies of the physiology, function and neural control principles of the mastication. Mastication is the action of breaking down of food, preparatory to deglutition. This breaking-down action is highly organized complex of neuromuscular and digestive activities. The duration and forces developed in the power stroke vary within and between individuals and for the type of the food being chewed. It has been suggested that the observation of masticatory movements may be of diagnostic value for assessing disorders of the stomatognathic system, but there is not clear evidence to show significant differences. The action of masticatory muscles during chewing varies between subjects in amplitude, onset timing, and duration of the chewing cycle. Since tooth guidance has an enormous influence on muscle activity during chewing and swallowing, it is advisable to make restorations compatible with the functional movement patterns of the patient rather than expect the patterns of the mastication to adapt to the new made restorations. PMID- 16340272 TI - Oral parafunctions and positive clinical examination findings. AB - STATEMENT OF PROBLEM: Oral parafunctional activity can be fatigued and painful masticatory muscles and/or pain in the temporomandibular joints. There is controversial discussion in the literature as to what role parafunctional activity plays in the multi-causal pathologic process. PURPOSE: The purpose of this study was to determine any association between the level of parafunctional habits versus the level of mandibular dysfunction and to test the hypothesis that TMD/bruxer patients have significantly increased muscle tension and joint pain. MATERIAL AND METHODS: There were 307 subjects (140 males und 167 females) selected for this investigation. 299 subjects were examined regarding any relationships between clenching and the incidence of muscle sensitivity. The age of subjects ranged from 20 to 54 years old, with a mean age of 35.4. 114 subjects had at least one sign of temporomandibular disorders (tenderness/pain on palpation of the joints or muscles, TMJ sounds, pain or deviation during maximum mouth opening (active/passive). RESULTS: 81 subjects admitted to clenching, while 218 said they did not. Among the "non-clenchers", 68.8% had no sensitive muscles, 31.2% indicated sensitive masticatory muscles. Those who clenched their teeth were distributed as follows: 53.1% were diagnosed with bilateral masticatory muscle sensitivity, 46.9% showed no such indications. The two groups were not homogeneously divided, with regard to pain/discomfort (p = 0.001; Fisher's precision test). CONCLUSION: This study found a statistically solid relationship between the incidence of "clenching" and muscle palpation findings, as well as between sensitivity in the mandibular joints from lateral and to cranial and dorsal with positive muscle palpation findings. The agreement between sensitivity of the masticatory musculature and the mandibular joint demonstrates that intensive clenching can predominantly lead to pathologic phenomena in the muscles or joints. PMID- 16340273 TI - Testing of the questionnaire on dental care satisfaction in a sample of adult patients visiting dental clinics at Faculty of Odontology, Kaunas University of Medicine. A pilot study. AB - A pilot study was conducted with the aim to test and develop a questionnaire which covers dental patients attendance, satisfaction with dental care, oral health self-evaluation, attitudes and knowledge about oral health, before applying it to a broad population. A total of 53 adult subjects (mean age 40.1) visiting Kaunas University of Medicine dental clinics, filled in the questionnaire presented by the researchers. Statement choices to define dental care satisfaction were selected from previously used instruments for similar studies in other countries. RESULTS: Major difficulties in answering questions were related to using the Likert scale response alternatives. All statements concerning dental care satisfaction with the mean Likert score value below 2.0 were decided to be indicative for the importance of characteristics. According to factor analysis and distribution of the mean scores of dental care satisfaction characteristics it was decided to include no more than 8 questions in each dimension to the final version of questionnaire. In case of equal loadings of several statements in the factor analysis, the statement with lower mean score was used. CONCLUSIONS: The use of professional terminology in questionnaire survey should be tested before applying to a broad population. The final instrument for measuring dental care satisfaction should be formed on the basis of statistical evaluation of relative importance and comparative load of different questions/statements to be included in the questionnaire. PMID- 16340274 TI - Secretory function of neutrophilic leukocytes of the patients with periodontal diseases. AB - Aim of study was to examine periodontal status among 20 44 year old patients and to study the secretory function of peripheral venous blood neutrophilic leukocytes (NL) exposed to various antigens and alkaline phosphatase (AP) activity in gingival crevicular fluid (GCF) in patients suffering from gingivitis and periodontitis. Clinically were determined Russell's periodontal index (PI). Secretory function of NL affected by opsonized zymosane, non-opsonized E. coli was examined in 77 patients with gingivitis and periodontitis, and in 35 donors, free of internal diseases, by means of beta-glucuronidase (beta-GD), lysozyme (LZ). NL secreted higher levels of beta-GD in incubation medium in patients with periodontitis (p < or = 0.001) subject to degree of periodontal lesion. NL affected by various antigens secreted higher levels of LZ into non-cellular matrix in patients with gingivitis and periodontitis comparing to control environment in analogous groups. Data obtained from this study suggest that in patients with periodontitis response of NL to bacterial stimuli is specific and subject to the degree of periodontal lesion. Our study showed a significant difference of AP activity in GCF subject to pocket depth and degree of periodontal lesion. Once NL are exposed to corpuscules prone to phagocytosis, an increase in secretion of beta-GD and LZ can be explained by overall increase in secretion of NL lysosomic enzymes, thus disclosing the mechanism of inflammatory periodontal tissue damage. PMID- 16340275 TI - 5 year experience with lower lip cancer. AB - Retrospective study of 189 cases of lower lip cancer treated from 1996-2000 is done. There were 69% males and 31% females. Median age was 66.8 years. 84.4% of patients were with tumors stage I-II. Surgical treatment was performed in 83.6% of patients. In all operated cases was squamous cell carcinoma as verrucous tumor in 17.4%, as exophytic in 46% and as ulcerative in 36.6%. There were different methods of local excision, primary reconstruction and neck dissection depending from stage. In the patient group with clinically negative neck at the first attendance (170 patients) delayed cervical metastases developed in 6 patients (3.5%). Recurrence at the primary site developed in 11.3% of patients and was associated with large tumor size and low cancer differentiation. Survival rate at 5-year follow-up was 95% for patients with I stage, 89.7% for II stage and 37% for III and IV stage patients or mean for all group 83.7%. Diagnosis and treatment of actinic cheilitis also is discussed. PMID- 16340276 TI - Assessment of antibacterial activity of some topical otological solutions. AB - OBJECTIVES: Otological solutions have long been used in the treatment of the bacterial and fungal infections of the ear. We investigated antibacterial activity of some otic solutions against the most common bacteria isolated from discharging ears. STUDY DESIGN: Three solutions were used (Castellani's, Burrow's, and 2% salicyl alcohol) for 20 fresh isolates of each of the following organisms: Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, Escherichia coli, and Enterobacter spp.. The activity of each solution was determined by the size of the zone of inhibition of bacterial growth. RESULTS: The Castellani's solution showed significantly larger average inhibition zones than the other solutions did (p<0.001). CONCLUSION: The Castellani's solution may be considered a good option against bacterial or mixed infections (bacterial and fungal) of the ear. PMID- 16340277 TI - [The relationship between skin test results and serum eosinophilic cationic protein, nasal eosinophilia, and total IgE values in patients with allergic rhinitis]. AB - OBJECTIVES: We investigated the relationship between positive skin test results and serum eosinophilic cationic protein (ECP), nasal eosinophilia, and total IgE levels in patients with allergic rhinitis. PATIENTS AND METHODS: The study included 36 patients with allergic rhinitis (26 females, 10 males; mean age 32.8 years; range 17 to 60 years) and with a positive skin test result. Serum ECP, nasal eosinophilia, and serum total IgE levels were assessed and compared with a control group of 18 healthy volunteers (15 females, 3 males; mean age 30.3 years; range 20 to 49 years) and correlation was sought with skin test results. RESULTS: Twenty-five patients were sensitive to only seasonal, four patients to only perennial, and seven patients to both seasonal and perennial allergens. Twelve patients were sensitive to one allergen, seven patients to two, and 17 patients to three or more allergens. Compared to controls, the mean serum ECP, nasal eosinophilia, and serum total IgE values were significantly higher in the patient group (p<0.05). These parameters did not show significant relationship with the type and number of allergens (p>0.05). CONCLUSION: Serum ECP, nasal eosinophilia, and serum total IgE levels increase in patients with allergic rhinitis. These parameters are not affected by sensitivity to seasonal or perennial allergens. PMID- 16340278 TI - [The prevalence of allergic rhinitis in the adult urban population of Denizli]. AB - OBJECTIVES: We investigated the prevalence of allergic rhinitis in the adult urban population of Denizli. STUDY DESIGN: A survey study was conducted to cover 1010 adults above 19 years of age, representing the urban population of Denizli district, Turkey. A questionnaire was designed to collect data on sociodemographic features, symptoms of allergic rhinitis, duration of symptoms, and any prior diagnosis of allergic rhinitis made by a physician. Interviews were made with randomly chosen adults from three socioeconomically different urban areas. RESULTS: Interviews were completed for 951 adults with ages ranging from 19 to 65 years (414 males, mean age 36.3+/-12 years; 537 females, mean age 35.2+/ 12 years). The frequency of self-reported allergic rhinitis was 10.8%, and 5.2% had allergic rhinitis diagnosed by a physician. Two or more symptoms of allergic rhinitis were found in 7.2%. The prevalence of allergic rhinitis was higher in females. There was no relationship between the frequency of allergic rhinitis and age. CONCLUSION: The prevalence of allergic rhinitis in the adult urban population of Denizli is between 5% to 10%, with a higher frequency in females. PMID- 16340279 TI - Radiofrequency for the treatment of vasomotor rhinitis. AB - OBJECTIVES: We assessed the efficacy of submucosal application of radiofrequency to the inferior turbinate for the treatment of vasomotor rhinitis. PATIENTS AND METHODS: Twenty patients with vasomotor rhinitis (9 males, 11 females; mean age 29.2 years; range 20 to 40 years) were treated with radiofrequency applied to the inferior turbinate. Symptoms such as nasal obstruction, sneezing, and watery nasal discharge were graded with the use of a visual analog scale (VAS) before, and on days 1, 3, 7, 30, 60, 90, and 180 after the treatment. RESULTS: The severity of symptoms began to decrease following the first week after the application. Maximum relief was achieved between 30 to 60 days after the intervention. The highest rate of improvement (85.4%) was reported in sneezing, followed by nasal obstruction (76.4%) and nasal discharge (67.7%). The mean VAS scores showed a significant improvement in all symptoms between 7 to 180 days after the procedure (p<0.05). The rate of patient satisfaction was 90% for the relief of nasal obstruction and sneezing, and 80% for nasal discharge. Complaints about vasomotor rhinitis increased up to a severity near the pretreatment level in eight patients on the 180th postoperative day and the procedure was repeated. CONCLUSION: These findings indicate that radiofrequency may be used as an alternative treatment option in patients with vasomotor rhinitis. PMID- 16340281 TI - [Pilomatrixoma of the head and neck in six cases]. AB - Pilomatrixomas are benign neoplasms of the hair follicle, presenting as a mass in the head and neck region. Those arising from the parotid region may pose a diagnostic challenge. We present six patients (age range 6 to 68 years) with pilomatrixoma in the head and neck region. The lesions were located in the preauricular region in two cases, and in the periorbital region, the forehead, the neck, and in the dorsum of the nose in the other patients, respectively. The main complaint was a hard, slow-growing mass in all the cases. All the patients were treated surgically. Histopathologic diagnosis was pilomatrixoma. No recurrences or surgery-associated complications were observed in the follow-up period. PMID- 16340280 TI - [The effect of combined use of vitamin C, vitamin E, and ibuprofen on flap viability: an experimental study]. AB - OBJECTIVES: Many studies have been carried out to investigate the individual effects of vitamin C, vitamin E, and ibuprofen on flap viability, with favorable results. This study aimed to determine the effect of combined use of these agents on flap viability. DESIGN AND METHODS: Sixty Wistar rats weighing 250-300 grams were divided into six groups, equal in number, to receive saline solution (group 1, control), vitamin C (group 2), vitamin E (group 3), vitamin C and E (group 4), ibuprofen (group 5), and vitamin C, vitamin E, and ibuprofen (group 6). Following ketamine anesthesia, a caudally based reverse McFarlane flap on the back of the rats, 3x10 cm in size, was elevated and sutured back. The agents were administered intraperitoneally once daily for seven days, after which viable flap areas were estimated. RESULTS: The percentages of the viable area of the flaps in groups 1, 2, 3, 4, 5, and 6 were 58.0%, 68.1%, 61.4%, 73.4%, 69.1%, and 80.5%, respectively. The use of vitamin C, vitamin E, and ibuprofen in combination resulted in a significantly greater flap viability compared to individual uses. CONCLUSION: Administering vitamin E and vitamin C together has a greater effect on flap viability than used alone; however, the benefit is the greatest with the addition of ibuprofen. PMID- 16340282 TI - Plasmacytoid myoepithelioma of the soft palate: a review of the literature and report of a case with immunohistochemical findings. AB - Myoepitheliomas of salivary glands are rare neoplasms, accounting for less than 1% of all salivary gland tumors. They are classified into four cell types: epithelioid, spindle, clear, and plasmacytoid. Among them, the plasmacytoid cell type appears to have a predilection for the oral cavity, especially the palate. We presented a 32-year-old man with plasmacytoid myoepithelioma arising in the soft palate, together with a brief discussion of the clinicopathologic features and differential diagnosis in the light of the literature. PMID- 16340283 TI - Radiologic findings of a congenital suprasternal dermoid cyst. AB - A congenital cystic mass was detected at the suprasternal notch of a seven-month old male infant. After radiologic examinations including ultrasonography, computed tomography, and magnetic resonance imaging, the cyst was excised and diagnosed as a dermoid cyst. Dermoid cysts of the head and neck are rare lesions, but a midline location is characteristic for these congenital masses. To our knowledge, only two reports have been published, which were similar to our case in localization. Dermoid cysts should be included in the differential diagnosis of midline cysts. PMID- 16340284 TI - Auditory neuropathy in children: diagnostic criteria and audiological test results. AB - OBJECTIVES: In this study, we evaluated pediatric patients with auditory neuropathy with regard to diagnostic criteria and audiological test results. PATIENTS AND METHODS: Hearing assessment was made in five children with auditory neuropathy (3 boys, 2 girls; age range 7 months to 6 years). The patients were tested with the use of acoustic immitance measures, transient evoked otoacoustic emissions (TEOAE), behavioral audiometry, and auditory brainstem responses (ABR). RESULTS: Transient otoacoustic emissions were recorded in all the patients in contrast to the lack of auditory evoked brainstem responses (i.e. there were no identifiable waves in all recordings). Another common feature was the absence of correlation between ABR, TEOAE, and behavioral test results. CONCLUSION: Of the above-mentioned measurements, otoacoustic emissions and the auditory brainstem responses, when used together, offer insight into preneural as well as neural function in the auditory system and thus, may form the necessary combination for the evaluation of hearing in children. PMID- 16340285 TI - [Expression of Ki-67 and p53 in laryngeal squamous cell carcinomas]. AB - OBJECTIVES: The aim of this study was to investigate the expression of p53 tumor suppressor gene and Ki-67 antigen in laryngeal squamous cell carcinoma and its relation to conventional prognostic indicators. PATIENTS AND METHODS: p53 and Ki 67 expressions were evaluated in surgical specimens of 37 patients (34 males, 3 females; mean age 57.2 years; range 35 to 75 years) with squamous cell carcinoma of the larynx. Immunohistochemical studies were performed in routinely fixed and paraffin-embedded tissue sections using the immunoperoxidase technique and monoclonal antibodies. RESULTS: The tumors were well-, moderately, and poorly differentiated in 5, 24, and 8 patients, respectively. Ten patients had lymph node metastasis, nine patients had vascular invasion, and 15 patients had neural invasion. Ki-67 and p53 nuclear staining was detected in 31 (83.8%) and 15 (40.5%) tumors, respectively. The incidence of Ki-67 expression was higher in patients with lymph node metastasis, but this did not exhibit significance. Neither p53 nor Ki-67 immunoexpression was in significant association with conventional clinicopathological parameters (p>0.05). CONCLUSION: These data indicate that p53 and Ki-67 are not correlated with conventional prognostic parameters in squamous cell carcinoma of the larynx. PMID- 16340286 TI - The effectiveness of unilateral tonsillectomy in chronic adenotonsillar hypertrophy. AB - OBJECTIVES: This study sought to determine whether unilateral tonsillectomy was effective in the treatment of chronic adenotonsillar hypertrophy. PATIENTS AND METHODS: A total of 197 patients (96 girls, 101 boys) with chronic adenotonsillar hypertrophy were prospectively included. The patients were randomly assigned to either bilateral tonsillectomy (n=113; mean age 10.1 years; range 4 to 18 years) or unilateral tonsillectomy (n=84; mean age 4.5 years; range 3 to 8 years) performed with or without adenoidectomy. All the patients were followed-up for at least a year. RESULTS: The two groups did not differ significantly with respect to preoperative symptoms, the size of tonsils, and the presence of adenoid hypertrophy. At the end of the follow-up, no significant differences were found with respect to relief of snoring, mouth breathing, and upper airways obstruction between the two groups. A greater number of patients achieved complete improvement in snoring in the bilateral tonsillectomy group (p<0.05). With unilateral tonsillectomies, three patients required tonsillectomy to the other side, two had recurrent tonsillitis, and one had severe compensatory hypertrophy with persistent difficulty in inspiration and mouth breathing. CONCLUSION: Unilateral tonsillectomy seems to be as effective as bilateral tonsillectomy in the treatment of chronic adenotonsillar hypertrophy. PMID- 16340287 TI - [Parental attitudes at the beginning of child's stuttering]. AB - OBJECTIVES: Stuttering is a communication disorder that frequently starts at the age of three or four years. Its appearance and progression may impose psychological effects on children and their families. We evaluated parental attitudes in the initial period of stuttering. PATIENTS AND METHODS: Twenty-two children or adolescents (16 males, 6 females; mean age 8.8+/-4.4 years; range 4 to 18 years) were assessed. A semi-structured clinical interview was administered to all the patients to determine whether stuttering was triggered by stressors and to examine the attitudes of parents and the course of their applying for treatment. RESULTS: After the onset of stuttering symptoms, the most frequent parental attitude was found as punishment and warning to their child (54.5%). Before stuttering appeared, the most frequent triggering event was of being excessively scared of anything (68.2%). The mean age of onset of stuttering was 3.4+/-1.1 years (range 2 to 7 years). CONCLUSION: The events that trigger stuttering also continue in the attitudes and thoughts of parents concerning stuttering. After the diagnosis, the parental attitude plays a significant role in the effectiveness of treatment. In this respect, parents should be informed during their first application for treatment. PMID- 16340288 TI - [Long-term results of intracordal or neocordal autologous fat injection in patients with unilateral vocal cord paralysis and cordectomy]. AB - OBJECTIVES: We investigated the long-term results of autologous fat injections for unilateral vocal cord paralysis and laryngofissure and cordectomy defects. PATIENTS AND METHODS: We evaluated the efficacy of autologous fat injections in 21 patients (16 males, 5 females; mean age 57 years; range 24 to 72 years) with unilateral vocal cord paralysis (n=11) and cordectomy defects (n=10) in the light of computed acoustic analysis, phonatory functions, and videolaryngostroboscopic findings and the effect of injections on swallowing. The mean follow-up was 14 months (range 10 to 22 months). RESULTS: Voice quality improved in 14 patients. Glottic insufficiency recurred in three patients in the early period, and in two patients in the late period. Phonatory functions showed significant improvement in all parameters except for jitter and functional interference (p<0.05). Videolaryngostroboscopy showed significant improvement in glottic closure, but mucosal waves remained unchanged. Of 18 patients with swallowing disorders, aspiration symptoms improved in 15 patients. Patients with unilateral vocal cord paralysis had better results compared to those with cordectomy defects. CONCLUSION: Autologous fat injections may be considered effective in patients with cordectomy defects or unilateral vocal cord paralysis. PMID- 16340289 TI - [Laser assisted eustachian tuboplasty: a case report]. AB - Surgical treatment of eustachian tube dysfunction is still challenging. Placement of a transtympanic ventilation tube may not be successful and may require multiple applications. We presented a 47-year-old male patient with chronic recurrent otitis media with effusion and retraction due to eustachian tube dysfunction, in whom the number of previous transtympanic tube insertions amounted to 18. He underwent laser assisted eustachian tuboplasty in both ears with endoscopic guidance under general anesthesia. The mucosal, submucosal, and partially the cartilaginous tissues in the posterior pillow of the nasopharyngeal orifice of the eustachian tube were ablated by Nd:YAG and holmium:YAG laser. Following surgery, air conduction thresholds decreased from 37 dB to 22 dB, and from 38 dB to 33 dB for the right and left ears, respectively. The preoperative tympanogram curve, which was of type B for both ears was found as type C-1 on the right side, and type C-2 on the left. Limited improvement seen in the left ear was ascribed to insufficient ablation of the posterior pillow of the eustachian tube. No complications were encountered within a follow-up of 12 months. PMID- 16340290 TI - Computed tomography findings of labyrinthitis ossificans secondary to meningitis: a case report. AB - A 26-year-old man was admitted to our hospital with chronic left ear drainage. He had a history of meningitis when he was a child. Computed tomography (CT) of the temporal bone showed complete obliteration of the otic labyrinth by sclerotic tissue. Based on CT findings, the patient was diagnosed as labyrinthitis ossificans. Computed tomography is an appropriate method of examination for the identification of labyrinthitis ossificans and is of particular importance for the evaluation of patients before cochlear implantation. PMID- 16340291 TI - Rhinocerebral mucormycosis: a report of two cases. AB - Rhinocerebral mucormycosis is an aggressive fungal disease that involves the nose, paranasal sinuses, orbit and central nervous system. It may rapidly be fatal. This infection usually occurs secondary to immune suppression, diabetic ketoacidosis, and prolonged use of antibiotics, steroids, and cytotoxic drugs. Management of the condition consists of treatment of the underlying disease and surgical debridement combined with intravenous amphotericin B. This paper presents two patients with rhinocerebral mucormycosis, both suffering from diabetes. Complete recovery was achieved in one patient with surgical debridement of necrotic tissue combined with systemic antifungal treatment, whereas in the other, the disease was complicated with subarachnoid hemorrhage and the patient died on the third day of treatment. PMID- 16340292 TI - A case of intranasal schwannoma with bilateral nasal polyposis. AB - Schwannoma is a benign neoplasm originating from schwann cells of the peripheral nerve sheath, and its occurrence in the nasal cavity and paranasal sinuses is rare. We present a case of nasal schwannoma originating from the left nasal cavity, accompanied by bilateral nasal polyposis. A 66-year-old man presented with complaints of progressive left nasal obstruction, hyposmia, and headache. Anterior rhinoscopic and endoscopic examinations revealed a mass that almost completely filled the left nasal cavity. A polypoid mass was present in the right nasal cavity, as well. On computed tomography, the mass occupied the left ethmoidal cells, left maxillary sinus, left sphenoid sinus, and posterior area of the left nasal cavity. Endoscopic sinus surgery was performed under general anesthesia and the mass was completely removed via the nasopharynx because of its size. Endoscopic anterior ethmoidectomy was also performed in the right nasal cavity for nasal polyposis. Histopathological diagnosis of the left nasal mass was schwannoma. The patient was symptom-free with no recurrence after nine months. PMID- 16340293 TI - [Evaluation of predisposing factors and bacteriologic agents in pediatric rhinosinusitis]. AB - OBJECTIVES: This study was conducted to investigate the predisposing factors and bacteriologic agents of acute rhinosinusitis in children. PATIENTS AND METHODS: The study included 76 patients (47 girls, 29 boys; mean age 10.2 years; range 4 to 18 years) with acute rhinosinusitis. Following a detailed history, the patients were investigated by otolaryngologic examination including nasal endoscopy, allergy tests, Water's graphy, and aspiration cultures from the middle meatal region. RESULTS: The leading complaints were nasal obstruction (92.1%), rhinorrhea (89.4%), and headache (51.3%). Allergy tests were positive in nine of 23 patients (30.3%) with a history of allergy. Concerning smoking, 39 patients (51.3%) had a secondary, two patients (2.6%) had a primary history. Sixteen patients (21.1%) had septal deviation, and 25 patients (32.9%) had benign adenoid tissue obstructing choanal spaces. Water's graphies showed pathology in 69 patients (93.2%). Cultures were positive in 51 patients (67.1%), with Streptococcus pneumoniae in 23 (41.1%), Haemophilus influenzae in 17 (30.4%), and beta-hemolytic streptococci in eight patients (14.2%). Treatment was comprised of ampicillin-sulbactam in 31 (40.8%), loracarbef in 20 (26.3%), and amoxicillin clavulanate in 13 (17.1%) patients. After a year follow-up, recurrence was detected in 12 patients (15.8%). No significant effect of the size of adenoid tissue and allergy was found on the success of treatment (p>0.05). CONCLUSION: Smoking, choanal obstruction by the adenoid tissue, anatomical variations, and allergy were the most frequent predisposing factors, while S. pneumoniae and H. influenzae were the commonest pathogens. PMID- 16340294 TI - [The predictive value of assessing mastoid pneumatization in pre-dive examinations]. AB - OBJECTIVES: We investigated the value of assessing the degree of mastoid pneumatization in predicting middle ear barotrauma -the most common problem in sport SCUBA divers- in comparison with that of the conventional pre-dive examination method, the Valsalva maneuver. MATERIALS AND METHODS: Thirty-four volunteer sport SCUBA divers having normal pre-dive examination findings were included. The Valsalva test was performed in all the divers. Mastoid pneumatization for each ear was calculated on a Schuller's view radiography. Pneumatization of = or <30 cm2 was accepted as poor (prone to barotrauma) and >30 cm2 was accepted as good (not prone to barotrauma). During the observation period, the divers were monitored with regard to barotrauma symptoms and signs. RESULTS: The degree of mastoid pneumatization was = or <30 cm2 in 26 ears (38%) of 16 divers (47%). The Valsalva test was negative in eight ears (12%) of six divers (18%). During a total of 1001 dives, symptomatic middle ear barotrauma occurred in 28 dives (2.8%), i.e. in 21 ears (31%) of 16 divers (47%). Of these, the degree of mastoid pneumatization predicted barotrauma in 11 (69%) divers, whereas the Valsalva test was negative in only three (19%) (p<0.05), with sensitivity, specificity, positive and negative predictive values, and efficiency being 69% and 19%, 72% and 83%, 69% and 50%, 72% and 54%, and 71% and 53%, respectively. CONCLUSION: The value of assessing mastoid pneumatization in predicting middle ear barotrauma in sport SCUBA divers is higher than that of the Valsalva test. PMID- 16340295 TI - [The value of laryngeal electromyography in the prognosis of vocal cord paralysis]. AB - OBJECTIVES: We investigated the value of laryngeal electromyography (EMG) in monitoring patients with vocal cord paralysis. PATIENTS AND METHODS: Cricothyroid and thyroarytenoid muscles of 40 patients (10 females, 30 males; mean age 32 years; range 18 to 61 years) with vocal cord paralysis were monitored with laryngeal electromyography. The patients were divided into two groups according to time to presentation after symptom onset, i.e. within 1-3 months (group 1; n=14), and after at least 12 months (group 2; n=26). Electromyography was repeated every three months in group 1, and in the third and sixth months in group 2. RESULTS: Initial EMG examinations showed polyphasic reinnervation potentials and partial axonal degeneration in 21 laryngeal nerves in group 1. Recovery was expected in these patients. Repeat EMGs revealed normal findings in 17 laryngeal nerves, yielding a prognostic estimation of 80.9% (17/21). In group 2, none of the patients demonstrated polyphasic potentials or motor unit potentials suggesting reinnervation on admission. The earliest and latest recoveries were observed in the fourth and eleventh months (mean 6.4 months) in group 1, respectively. However, group 2 patients were followed-up for at least 18 months after the onset of their symptoms and none manifested clinical and electrophysiologic improvement. CONCLUSION: Our data suggest that EMG is a useful technique in the estimation of prognosis of patients presenting in the early period of vocal cord paralysis. PMID- 16340296 TI - [Evaluation of the protective effect of magnesium on amikacin ototoxicity by electrophysiologic tests in guinea pigs]. AB - OBJECTIVES: This experimental study was performed to evaluate the protective effect of magnesium on amikacin ototoxicity. STUDY DESIGN: Twenty-seven guinea pigs (54 ears) with normal auropalpebral reflexes and otomicroscopic examination, auditory brain stem responses (ABR), and transient evoked otoacoustic emissions (TEOAE) were randomly divided into four groups to receive amikacin once 15 mg/kg daily (group 1 and 3) and twice 7.5 mg/kg daily (group 2 and 4) for seven days. Groups 3 and 4 were administered oral MgCl an hour after amikacin use at a dose of 39 mmol/l and 19.5 mmol/l, respectively, for seven days. Otomicroscopic examination, ABR and TEOAE measurements were repeated on the third, fifth, and seventh days. RESULTS: Compared to group 1, decreases in TEOAE amplitudes and reproducibility and increases in ABR thresholds were significant in group 2 (p<0.05). However, in groups 3 and 4, TEOAE amplitudes remained unchanged, or even increased and the reproducibility of TEOAE responses and ABR thresholds showed no significant changes (p>0.05). CONCLUSION: Our data show that oral magnesium may play a protective role on amikacin ototoxicity. PMID- 16340297 TI - Simultaneous existence of papillary carcinoma in the thyroid gland and thyroglossal duct cyst in two patients. AB - Papillary carcinoma of the thyroglossal duct cyst is a rare occurrence. Two patients presenting with medial neck masses were diagnosed as having thyroglossal duct cysts by ultrasonography. The Sistrunk operation was performed. Histopathologic evaluation demonstrated papillary carcinoma in the surgical specimens of both patients. The thyroid glands were examined by ultrasonography, scintigraphy, and fine-needle aspiration biopsy. Biopsy showed papillary carcinoma, and total thyroidectomies were performed. Micropapillary carcinoma was detected in the resected thyroid glands. The patients were asymptomatic without complications after 24 and 32 months of follow-up, respectively. PMID- 16340298 TI - [Dermatofibroma accompanied by perforating dermatosis in the auricle: a case report]. AB - Although dermatofibroma is one of the most common soft tissue tumors, it is rarely seen in the face. Perforating dermatosis is characterized by papulonodules measuring 1-10 cm, with a central cup-shaped epidermal depression filled with a keratotic plug. An 82-year-old woman presented with a hyperkeratotic erythematous papular lesion, 0.8 cm in diameter, on the posterior side of the left auricle, showing no infiltration to the peripheral tissues. Total excisional biopsy was performed, which showed dermatofibroma accompanied by perforating dermatosis. No recurrence was detected during her follow-up. Our literature search did not yield any reported case of dermatofibroma accompanied by perforating dermatosis. PMID- 16340299 TI - [Three cases of mucosal malignant melanoma of the nose and paranasal sinuses]. AB - We evaluated the clinical course and treatment of three patients who were found to have mucosal malignant melanoma of the nose and paranasal sinuses. The presenting complaint was recurrent epistaxis in all the patients. One patient had been followed-up for nasal polyposis for a long time. In older patients with recurrent epistaxis, endoscopic examination of the nasal cavity and obtaining a biopsy from suspected regions are essential for early diagnosis of mucosal malignant melanomas. PMID- 16340300 TI - Vertigo and medico-legal problems. AB - Forensic medicine deals with the interactions of medical science with the law. All branches of medicine can play a role in dealing with medico-legal problems. Clinical and forensic evaluators need to be able to recognize variations of symptoms or syndromes that "don't read the textbooks". Atypical presentations of dizziness or vertigo have the potential to cause much confusion in diagnosis, treatment planning, and legal adjudication, if not correctly recognized. This article describes the forensic medicine in the legal system and forensic aspects of other medical specialities, gives an overview about medico-legal viewpoint of vertigo, and underlines the importance of objective clinical and forensic evaluation of the patient with vertigo. PMID- 16340301 TI - A survey of Enterobius vermicularis infection among children on western and southern coastal islands of the Republic of Korea. AB - To determine the status of Enterobius vermicularis infection among children living on western and southern coastal islands of the Republic of Korea, children (3-10 years) in 39 kindergartens and primary schools were examined using the cello-tape anal swab method, during July and August 2000. Of 1,661 children examined, 307 (18.5%) were found to be positive for E. vermicularis eggs. The highest infection rate (59.3%) was found in a kindergarten and a branch school of Shinyang primary school on Chujado, Jeju-do (Province). Remarkable differences in egg positive rates were observed in different localities. The egg positive rate for boys (21.3%) was significantly higher than that of girls (15.4%) (P = 0.02). However, positive rates were not significantly dependent on age. The results of the present study show that E. vermicularis infection is highly prevalent among pre-school and primary school children living on the western and southern coastal islands of the Republic of Korea, and indicate the need for efforts to control this infection. PMID- 16340302 TI - A seroepidemiological survey of Taenia solium cysticercosis in Nabo, Guangxi Zhuang Autonomous Region, China. AB - We have observed the seropositive rate of Taenia solium cysticercosis in residents at Nabo Village, Tiandong County, Guangxi Zhuang Autonomous Region, China by enzyme linked immunosorbent assay. The village had been found to be a relatively high endemic area of porcine cysticercosis among roaming pigs. Of 202 persons examined four males aged 15, 25, 35 and 41 year-old exhibited absorbance (abs) at 0.18, 0.20, 0.35 and 0.55, respectively. In addition, two females whose ages were 35 and 39 years revealed specific antibody levels of abs 0.26 and 0.41 in their sera. Overall positive rate among the people was 2.97%. All of these persons agreed that they had ingested the pork infected with T. solium metacestode (TsM), while history of proglottid discharge was not noticed from all of them. Three males and one female complained of intermittent headache. Our findings reinforced not only that the prevalence of cysticercosis might be related with roaming pigs infected with TsM but also that behavioral and environmental practices in local community constituted risk factors for transmission of the infection. PMID- 16340303 TI - A new culture system for in situ observation of the growth and development of Eucyclops serrulatus (Copepoda: Cyclopoida). AB - A practical and convenient method of rearing Eucyclops serrulatus in a microculture environment is described. A complete life cycle of E. serrulatus was maintained in a narrow space on a microscope slide glass on which a cover glass of 22 x 40 mm in size was mounted at a height of 0.8 mm. The culture medium was constituted by bottled mineral water boiled with grains of Glycine max (soybean). Chilomonas paramecium, a free-living protozoan organism, was provided as live food. Growth of nauplii hatched from eggs to the first stage of copepodite took an average of 7.7 days, and the growth of copepodite 1 to the egg-bearing adult female took an average of 20.1 days in the microculture cell with an average life time of 44.7 days. Continuous passage of copepods was successfully maintained as long as sufficient medium and food were provided. The microculture method enables an in situ microscopic observation on the growth and developmental process of helminth larvae experimentally infected to copepods as well as of copepod itself. Furthermore, it does not require anesthetization and, therefore, minimize the amount of stress exposed to copepods during the handling process. PMID- 16340304 TI - Expressed sequence tags analysis of Blattella germanica. AB - Four hundred and sixty five randomly selected clones from a cDNA library of Blattella germanica were partially sequenced and searched using BLAST as a means of analyzing the transcribed sequences of its genome. A total of 363 expressed sequence tags (ESTs) were generated from 465 clones after editing and trimming the vector and ambiguous sequences. About 42% (154/363) of these clones showed significant homology with other data base registered genes. These new B. germanica genes constituted a broad range of transcripts distributed among ribosomal proteins, energy metabolism, allergens, proteases, protease inhibitors, enzymes, translation, cell signaling pathways, and proteins of unknown function. Eighty clones were not well-matched by database searches, and these represent new B. germanica-specific ESTs. Some genes which drew our attention are discussed. The information obtained increases our understanding of the B. germanica genome. PMID- 16340305 TI - Partial characterization of a 29 kDa cysteine protease purified from Taenia solium metacestodes. AB - A 29 kDa cysteine protease of Taenia solium metacestodes was purified by Mono Q anion-exchanger and Superose 6 HR gel filtration chromatography. The enzyme was effectively inhibited by cysteine protease inhibitors, such as iodoacetic acid (IAA) and trans-epoxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) while inhibitors acting on serine- or metallo-proteases did not affect the enzyme activity. The purified enzyme degraded human immunoglobulin G (IgG), collagen and bovine serum albumin (BSA), but human IgG was more susceptible for proteolysis by the enzyme. To define the precise biological roles of the enzyme, more detailed biochemical and functional studies would be required. PMID- 16340306 TI - Two species of goby, Boleophthalmus pectinirostris and Scartelaos sp., as the new second intermediate hosts of heterophyid fluke in Korea. AB - A survey was performed in order to determine the infection status of the metacercariae of heterophyid fluke in two goby species, Boleophthalmus pectinirostris and Scartelaos sp., collected from Gangjin-gun, and Shinangun, Sooncheon-shi, Jeollanam-do, Republic of Korea. A total of three metacercariae of Heterophyopsis continua was found in only one B. pectinirostris (10.0%) from Gangjin-gun. Heterophyes nocens metacercariae were detected in 24 B. pectinirostris (96.0%) and 14 Scartelaos sp. (63.6%) from Shinan-gun. Heterophyopsis continua metacercariae were found in 11 B. pectinirostris (44.0%) and 21 Scartelaos sp. (95.5%) from Shinan-gun. Stictodora fuscata metacercariae were detected in 18 B. pectinirostris (72.0%) from Shinan-gun. No metacercariae were detected in 20 B. pectinirostris from Sooncheon-shi. From the above results, this study is the first to prove that B. pectinirostris and Scartelaos sp. serve as the second intermediate hosts of some heterophyid flukes in Korea. PMID- 16340307 TI - Glucocorticoids and progression of breast cancer. PMID- 16340308 TI - A plasmid-encoded VEGF siRNA reduces glioblastoma angiogenesis and its combination with interleukin-4 blocks tumor growth in a xenograft mouse model. AB - Angiogenesis is required for the development and biologic progression of glioblastoma multiform (GBM), which is the most malignant infiltrative astrocytoma. Vascular endothelial growth factor (VEGF) plays a predominant role in the increased vascularity and endothelial cell proliferation in GBMs driven by the expression of pro-angiogenic cytokines. In this study, we employed a vector encoded VEGF siRNA to impair VEGF secretion from U87 human glioblastoma cells. The direct intra-tumor injection of a siRNA-encoding plasmid complexed with linear polyethylenimine (PEI) efficiently reduced the vascularization of treated tumors in xenografts established in SCID mice by subcutaneous inoculation of U87 cells, but was not able to reduce tumor growth. We then sought to strengthen the in vivo action of our siRNA by coupling it to a well known direct antiangiogenic agent, mouse interleukin 4 (mIL4). We infected U87 cells with a retroviral vector coexpressing the VEGF siRNA and mIL4 and produced stable cell lines that we used for an in vivo experiment of subcutaneous injection in SCID mice. In this setting, the concomitant expression of mIL4 and siRNA totally abolished the growth of subcutaneous tumors. These results suggest that our retroviral vector might be employed as a potential tool in future antiangiogenic gene therapy trials for glioblastoma. PMID- 16340309 TI - E2F3a stimulates proliferation, p53-independent apoptosis and carcinogenesis in a transgenic mouse model. AB - Mutation or inactivation of the retinoblastoma (Rb) tumor suppressor occurs in most human tumors and results in the deregulation of several members of the E2F family of transcription factors. Among the E2F family, E2F3 has been implicated as a key regulator of cell proliferation and E2F3 gene amplification and overexpression is detected in some human tumors. To study the role of E2F3 in tumor development, we established a transgenic mouse model expressing E2F3a in a number of epithelial tissues via a keratin 5 (K5) promoter. Transgenic expression of E2F3a leads to hyperproliferation, hyperplasia and increased levels of p53 independent apoptosis in transgenic epidermis. Consistent with data from human cancers, the E2F3a transgene is found to have a weak oncogenic activity on its own and to significantly enhance the response to a skin carcinogenesis protocol. The phenotype of K5 E2F3a transgenic mice is distinct from similar transgenic mice expressing E2F1 or E2F4. In particular, E2F3a has a unique apoptotic activity and lacks the tumor suppressive property of E2F1 in this model system. PMID- 16340310 TI - Contrast-enhanced in vivo imaging of breast and prostate cancer cells by MRI. AB - The development of effective cancer therapies has been hampered, in part, by the inability to noninvasively follow tumor progression from the initial cancerous lesion through to metastasis. We have previously shown that superparamagnetic iron oxide particles can be used as magnetic resonance imaging contrast agents to label embryonic, mesenchymal and hematopoietic stem cells in vivo. Improving the capacity to non-invasively image cancer progression is an appealing method that could be useful for assessing the efficacy of anticancer therapies. We have established that human prostate (LNCaP, DU145, PC3), rodent prostate (TRAMPC1, YPEN-1), human breast (MDA-MB-231) and mouse mammary (Myc/VEGF) cancer cell lines were readily labeled by fluorescent superparamagnetic sub-micron particles of iron oxide (MPIOs). The MPIOs were essentially inert with respect to cell proliferation and tumor formation. Fluorescence stereomicroscopy and three dimensional magnetic resonance imaging (MRI) determined that subcutaneous, intramuscular or orthotopically implanted labeled cancer cells could be imaged, in vivo, despite in some cases being undetectable by manual palpation. The MPIO labeled cancer cells could also be imaged, in vivo, at least 6 weeks after implantation. The fluorescent MPIOs further allowed for the ex vivo identification of tumors cells from histological sections. This study demonstrates the feasibility of using fluorescent MPIOs in prostate and breast cancer cell lines as both a negative contrast agent for in vivo MRI as well as a fluorescent tumor marker for optical imaging in vivo and ex vivo. PMID- 16340311 TI - Anaphase topsy-turvy: Cdk1 a securin, separase a CKI. AB - Chromosome segregation in mitosis and meiosis is triggered by activation of a large protease, separase. While it has been known for some time that the anaphase inhibitor securin regulates separase activity recent work shows that this is only half the story. In vertebrates Cdk1-dependent inhibition of separase represents a second, securin-independent branch of anaphase regulation. Furthermore, an unanticipated ability of separase to inhibit Cdk1 suggests additional, nonproteolytic functions of separase. PMID- 16340313 TI - Making performance reports work. AB - Public performance reporting is an increasingly common health policy tool to support accountability and quality improvement but there are few formal evaluations of this trend. In this review, we suggest that performance reporting may be an effective way to support improvements in performance when it is directed towards groups of providers rather than individuals and that there is enough evidence to support the use and further development of public performance reporting. However, the true impact of performance reporting depends on the policy context around reporting including clarity of strategy, incentives, and support for performance improvement. PMID- 16340314 TI - Back to basics on report cards. AB - Making Performance Reports Work is a valuable and timely contribution to the literature, particularly at the macro level when First Ministers have made commitments in 2000, 2003 and 2004 to report to their respective populations on a series of indicators, including wait times. PMID- 16340315 TI - Extreme makeover: can we achieve rapid improvement in Canada's healthcare system? AB - Building upon some key discussion points in the Brown et al. paper, we explore the key elements driving performance measurement and quality improvement strategies in the Veterans Affairs healthcare system in the United States and the national primary-care trusts in England, both of which offer important insights into understanding the factors that affect rapid, large-scale change. In the context of these "extreme makeover" examples, our commentary discusses the currently evolving performance measurement culture in the Canadian primary healthcare reform setting. We specifically highlight the experiences in Saskatchewan, a province that has been acknowledged recently by CIHI as a leader in primary healthcare evaluation. Although Saskatchewan has attempted to overcome the methodological and conceptual challenges in evaluation that Brown et al. outline in their paper, a stable performance measurement culture has yet to emerge and systematically utilize performance measurement reports for purposes of facilitating change. Although there is a growing recognition that measures by themselves will not be able to spur improvement, it is yet to be seen to what extent these performance reports can speak compellingly to policymakers, primary healthcare providers and managers to serve as catalysts to a major leap forward in overall quality improvement. PMID- 16340316 TI - Why performance indicators have a place in health policy. AB - Despite the mixed results of Brown and colleagues' review of the evidence for the use of performance indicators in health policy, this paper argues that they have an important place. Healthcare organizations cannot rely on altruism alone to motivate improved performance. Berwick, supporting the use of performance indicators in healthcare, argues "threats to survival are necessary to build will for improvement." He argues that the job of managers is to create organizations where such threats are clearly perceived, but balanced against a culture of "safety" in which individuals can learn and improve the care provided. This step in the causal chain gets insufficient attention from the "KAB+" evaluation model that Brown and colleagues employ. The use of performance indicators in publicly funded healthcare systems goes beyond arming the "consumer" of healthcare with relevant information; it is a fundamental question of democratic accountability. Methods for evaluating the evidence base of public policy must take into account the contextual (economic, social and political) factors that support or impede the achievement of policy objectives. PMID- 16340317 TI - Target and tailor the data. AB - Brown et al. provide a comprehensive evaluation of the public reporting of healthcare performance data, with, however, some limitation on recommendations for its effectiveness. Experience gained over the past 20 years is predominantly in the US with some in the UK. The small amount of Australian experience is reported in this paper. Accountability appears to override quality improvement as the purpose of publishing. It is easier to recount negative effects than positive ones but the overriding finding is the relatively small impression public disclosure has had on healthcare delivery. In an effort to overcome this problem, the intended audience should be clarified and the information correspondingly tailored. For consumers, information should mainly be concerned with issues of process and patient satisfaction, and for the providers, predominantly outcome data. PMID- 16340318 TI - Making performance reports work: it's a matter of common sense. AB - In the paper Making Performance Reports Work by Brown, Bhimani and MacLeod, the authors based their assessment of existing pubic performance reports on the Knowledge, Attitudes and Behaviour (KAB) model, with the addition of performance or actual healthcare outcomes to assess overall impact on the delivery system (i.e., the KAB+performance model). In short, the authors logically conclude that public performance reports about the healthcare delivery system are not effective unless patients ultimately use them, and/or unless providers improve their performance because of them. PMID- 16340319 TI - Performance reporting: alignment with accreditation. AB - This paper reviews the strong relationship of performance measurement and reporting with the Canadian Council on Health Services Accreditation (CCHSA) accreditation program and consistency with the direction of ongoing development. The CCHSA authors discuss the issue of public reporting of the accreditation results, the impact of public reporting on accreditation, aligning accreditation with the public reporting requirements, the measurement framework, quality improvement results and reporting and the linkage of accreditation with accountability. As public reporting gains momentum, the accreditation program must align to enable this while protecting the fundamental integrity and value of the program. Participating in the accreditation program clearly demonstrates the organization's commitment to accountability. PMID- 16340320 TI - Public reporting makes accountability possible. AB - This paper reviews the meta-analysis of Brown, Bhimani and MacLeod and finds it an important contribution to the literature on performance reporting (PR) in healthcare. Of 1,053 papers identified in their search, only 62 met their criteria for inclusion in the review. The authors used a Knowledge, Attitudes and Behaviour (KAB) model for their analysis. Of the 63 papers reviewed, most related to individual consumers and groups of providers, fewer related to groups of consumers and individual providers. The principal findings were that there is a shortfall in the literature on PR to 2004 and limited evidence that PR is having any impact on KAB of consumers or providers. They appropriately conclude that there is a need for the development of a conceptual model and thereby the requirement that the goal of placing PR "within a context that focuses attention and that supports action" may be achieved. In general, the conclusions are defensible and, given the lack of impact of PR they have demonstrated, their work should stimulate response and action within healthcare systems. Finally, this paper concludes by proposing six characteristics of successful PR--clear definition of context and goals, objective and verifiable indicators, transparent and useable information, equitability in scope, engaging the continuum of care, and integrating measures of cost-utility. PMID- 16340321 TI - Determining the utility of public reporting--too early to judge. AB - This paper presents several hypotheses about why public reporting of performance information in healthcare has not had more impact. Abstracting information from paper-based systems is slow and expensive, impairing the ability to provide timely, accurate information about system performance. Alternatively, the incentives that drive the behaviour of participants in the system may obstruct meaningful change in response to performance information. The delivery of healthcare is a very complex enterprise. This makes creating a set of indicators that is both useful and readily understandable by the general public very difficult. Perhaps the industry should consider the development of composite indicators not unlike those used to report on the macroeconomic performance of regional economies. In this regard, the development of composite indicators for quality of care (using measures of evidence-based protocol adherence) and access (through wait-times measures) is suggested. In conclusion, the paper states that it is too early to judge the efficacy of public reporting of performance information in healthcare; much more development of performance reporting is required. PMID- 16340322 TI - About context and systems. AB - This commentary on the paper of Adalsteinn Brown et al. addresses the various tensions between global evidence, general theories and local change processes. Following up on this analysis, the paper also discusses the impact of public performance data on both consumer behaviour and provider behaviour. The four lessons presented by Brown et al. are put in the context of considering the public release of performance data as a social process and an integral part of the Canadian health system design and functioning. PMID- 16340323 TI - The importance of context and attention. PMID- 16340325 TI - Can you prevent ventilator-associated pneumonia? PMID- 16340327 TI - The latest in trauma care. PMID- 16340328 TI - The latest in cardiac care. PMID- 16340329 TI - The latest in respiratory care. PMID- 16340331 TI - Outmuscling rhabdomyolysis. PMID- 16340333 TI - Neurocysticercosis: evaluation with advanced magnetic resonance techniques and atypical forms. AB - Neurocysticercosis (NCC) is the most common helminthic infection of the central nervous system, but its diagnosis remains difficult. The purpose of this article is to perform a critical analysis of the literature and show our experience in the evaluation of NCC. We discuss the advanced MR technique applications such as diffusion and perfusion-weighted imaging, spectroscopy, cisternography with FLAIR, and supplemental O2 and 3D-CISS. The typical manifestations of NCC are described; emphasis is given to the unusual presentations. The atypical forms of neurocysticercosis were divided into: intraventricular, subarachnoid, spinal, orbital, and intraparenchymatous. Special attention was also given to reactivation of previously calcified lesions and neurocysticercosis associated with mesial temporal sclerosis. PMID- 16340334 TI - Imaging aspects of pyogenic infections of the central nervous system. AB - Although pyogenic infections of the central nervous system are not a frequent group of diseases, their morbidity and mortally are very high. For this reason they require prompt diagnosis and treatment to avoid several complications that can lead to an undesired outcome. In this article, we review the imaging findings of these infections according to the anatomic site, their complications, and their differential diagnosis. Special attention is given to the different techniques of magnetic resonance imaging like perfusion, spectroscopy, and diffusion, for each specific situation such as meningitis, abscess, ventriculitis, purulent extra axial collections, and vascular complications. PMID- 16340335 TI - Granulomatous diseases of the central nervous system. AB - Infectious diseases of the central nervous system (CNS), particularly those accompanied by the formation of granulomas, are a constant diagnostic challenge in some specific regions of the world, above all in developing countries. The pattern of image seen on CT or MR scan is the result of the inter-relations between the individual characteristics of the infectious agent and the capacity of each host to mount an appropriate inflammatory response to that specific type of aggression, inside one particular compartment of the CNS. Taking these parameters into account we will discuss the several patterns of image found in parasitic, bacterial, and fungal granulomatous infections. PMID- 16340336 TI - Viral diseases of the central nervous system. AB - Viral diseases of the central nervous system encompass a wide range of different processes, mainly inflammation affecting the brain (encephalitis), the meninges (meningitis), or a combined meningoencephalitis. The spinal cord can be affected as well (myelitis). Another group of viral-related disorders, sometimes without a clear pathophysiological mechanism disclosed, include post-viral illnesses. All of these groups of diseases are discussed in this article, with an emphasis on their imaging presentation, using magnetic resonance imaging. PMID- 16340337 TI - Subacute spongiform encephalopathies. AB - Subacute spongiform encephalopathies are rare fatal diseases that affect the central nervous system, which is thought to be caused by prions, characterized clinically by a rapid progressive dementing course, along with generalized myoclonus. The prototype of these conditions in humans is Creutzfeldt-Jakob Disease (CJD). Although the final diagnosis depends on neuropathological examination, the presence of periodic sharp wave complexes on EEG and of the neuron-specific enolase, tau protein, S-100, and of the 14-3-3 protein in the cerebrospinal fluid, make the diagnosis of probable CJD. However, as these criteria are not completely accurate and the early diagnosis is extremely difficult, much interest has been focused recently on imaging methods. With the advent of diffusion-weighted imaging, MRI has shown high sensitivity and specificity, therefore being considered a useful method for the early diagnosis of this entity. PMID- 16340339 TI - Pediatric mechanical circulatory support. AB - Extracorporeal membrane oxygenation (ECMO) is based on technology that has been used clinically for decades. Largely because of its widespread use in pediatric patients with respiratory failure, familiarity with this technology has established ECMO as the most commonly used form of circulatory support for children. However, clinical applications and technical aspects of ECMO support continue to evolve, ensuring the ongoing importance of this modality for pediatric circulatory support. Although traditionally lagging behind development for adult applications, the use of ventricular assist devices (VADs) is expanding in pediatrics. Of particular interest, a number of new VADs designed specifically for children are becoming available, while early development of some innovative devices for pediatric circulatory support is currently underway. PMID- 16340342 TI - Latest developments in the field of assisted circulation. AB - The 51st Annual Meeting of the American Society for Artificial Internal Organs was held in Washington, DC, June 9 to 11, 2005. The abstract submission deadline for this meeting occurred on January 14, 2005. To ensure that the attendees were completely up-to-date with developments in the field of mechanical circulatory support, I was asked to provide a brief summary of developments or milestones that occurred after the abstract submission deadline. The list of highlights included in this manuscript was selected from an informal poll of colleagues and industry representatives, as well as a review of press releases and government- and industry-sponsored Web sites. Any omissions or errors in this summary of the latest developments in the field of mechanical circulatory support are entirely my responsibility. I also want the reader to understand that I have no financial interest in any of the device manufacturing companies to which I will refer. PMID- 16340343 TI - Post hospital care of the permanent LVAD patient. AB - Adrian Banning, consultant cardiologist, and Peter Houghton, long-term left ventricular assist device (LVAD) wearer and psychotherapist, reflect on their 5 year relationship in helping Peter adjust to a life relying on an LVAD and ask what the experience teaches us about the use of LVADs as a long-term palliation for heart failure. The article notes the ups and downs of maintaining health for LVAD wearers, how they respond to common complaints like colds, infections, and so on, and what monitoring is vital both to check general health and pump performance. Some of the psychological problems of what the authors call "extra life" are also noted. The article is not solely based on the experience of the two presenters but also on their work with five other implantees, of whom three have died from unrelated problems (e.g., cancer and an accident). PMID- 16340347 TI - Technology in clinical practice. AB - Salt and water are the major components of extracellular fluid, and each patient on dialysis has a unique serum sodium activity that is defended to ensure homeostasis. Dialysate use that does not restore extracellular composition to its equilibrium value results in disequilibrium and affects dialysis discomfort, blood pressure, thirst, and weight gain. When dialysate sodium activity exceeds the dialyzable plasma sodium activity, sodium diffuses from the dialysate into the patient. This results in hypertonicity, cellular dehydration, plasma volume expansion, and hypertension. A dialysate sodium activity below the patient's dialyzable plasma sodium reverses this process. Neither of these situations is desirable. Rather, dialysis should restore homeostasis by reconstituting the extracellular fluid to its desired salt and water content. Because each patient has a unique serum sodium activity, this goal can only be realized by using individualized dialysate sodium prescriptions. Further, because a patient's dialyzable sodium is dynamic and determined by serum chemistry plus dialyzer membrane behavior, the dialysate sodium needed to achieve isonatemic dialysis can only be defined by measuring the dialyzable serum sodium activity during dialysis. Achieving balanced salt and water removal to preserve homeostasis and intracellular tonicity requires automated sensors and feed-back control of dialysate conductivity. PMID- 16340348 TI - MagScrew TAH: an update. AB - The MagScrew Total Artificial Heart (TAH) system is the result of a close collaboration among the Cleveland Clinic Foundation, Foster Miller Technologies, Wilson Greatbatch Ltd, and Whalen Biomedical Inc. The system components are the thoracic blood pumping unit with attached compliance chamber and refill port, implantable electronic control unit, implantable battery pack, transcutaneous energy transmission system, external battery pack, and a telemetry system for communication with the electronic control unit. System in vitro tests are underway for system characterization and durability demonstration, whereas in vivo tests were conducted to evaluate system performance and biocompatibility under physiologic conditions. The passively filling pump uses a left master alternate left and right ejection control mode and has a Starling law-like response to venous pressure. The in vitro tests documented excellent hydraulic pump performance with high device output of over 9 l/min at left atrial pressures below 12 mm Hg. Atrial balance was well maintained under all test conditions. The in vivo tests demonstrated good biocompatibility without use of anticoagulant therapy. Experimental durations have ranged between 0 and 92 days. Postexplant evaluation of tissue samples did not reveal any sign of thromboembolic events or tissue damage due to device operation. PMID- 16340349 TI - Flow visualization study of a pulsating respiratory assist catheter. AB - Our group is currently developing an intravenous respiratory assist device that uses a centrally located pulsatile balloon within a hollow fiber bundle to enhance gas exchange rate via active mixing mechanism. We tested the hypothesis that the non-symmetric inflation and deflation of the balloon lead to both nonuniform balloon-generated secondary flow and nonuniform gas exchange rate in the fiber bundle. The respiratory catheter was placed in a 1-in. internal diameter rigid test section of an in vitro flow loop (3 L/min deionized water). Particle image velocimetry (PIV), which was used to map the velocity vector field in the lateral cross-section, showed that the balloon pulsation generated a nonuniform fluid flow surrounding the respiratory assist catheter. PIV was also used to characterize the fiber bundle movement, which was induced by the balloon pulsation. Gas permeability coefficient of the device was evaluated by using both the fluid velocity and the relative velocity between the fluid and the fiber bundle. The highest difference in the gas permeability coefficient predicted by using the relative velocity was about 17% to 23% (angular direction), which was more uniform than the 49% to 59% variation predicted by using the fluid velocity. The movement of the fiber bundle was responsible for reducing the variation in the fluid velocity passing through the bundle and for minimizing the nonuniformity of the gas permeability coefficient of the respiratory assist catheter. PMID- 16340350 TI - Performance and reliability of the CPB/ECMO Initiative Forward Lines Casualty Management System. AB - The Cleveland Clinic Foundation CPB/ECMO Initiative Forward Casualty Management System is an economical, compact, transportable, disposable system designed to permit a rapid expansion of trauma management services requiring cardiopulmonary bypass (CPB) or extracorporeal membrane oxygenation (ECMO) pulmonary support. The system, composed of a rotary blood pump, a pump motor driver, and an electronic control console as the blood pumping subsystem, also includes commonly used compatible commercial oxygenators, venous reservoirs, and cannulae. In vitro durability testing accumulated over 100 hours without failure. In vivo reliability was tested in 10 calves under general anesthesia during 6 hours of CPB and ECMO under full heparinization at nominal operating conditions of 4-5 l/min and 2-4 l/min blood flow respectively, and mean arterial pressures between 65 and 100 mm Hg. A mean time to failure of 57 hours was reached during the animal series. Results of these test series demonstrated that this system has the capability to reliably operate during a 6-hour conventional CPB or ECMO procedure, while providing flexibility and ease of use for the operator. PMID- 16340351 TI - In vitro performance of the novel coronary sinus AutoRetroPerfusion Cannula. AB - Myocardial salvage through coronary sinus intervention has been documented. The AutoRetroPerfusion Cannula is a novel device that is able to perfuse the coronary bed retrogradely through the coronary sinus with arterial blood generated from a peripheral artery with no need for a pump. The cannula consists of a distal end that, once secured in the coronary sinus, opens an umbrella-like membrane to create pressure in the coronary sinus, and at the same time has small channels directed backwards to the right atrium to provide pressure relief. The cannula is introduced from the axillary vein under local anesthesia and the proximal end, which consists of a graft, is anastomosed to the axillary artery to start autoperfusion once the distal end is secured in the coronary sinus and the occluding membrane is open. The AutoRetroPerfusion Cannula was tested in the in vitro mock loop under 50-120 mm Hg of proximal pressure and 50, 100, and 150 ml/min of total flow in the cannula. We were able to achieve the nominal design point of 40-80 mm Hg of distal pressure and 50-150 ml/min of distal flow by adjusting the number, diameter, and length of the small backwards channels. PMID- 16340352 TI - Development of a novel intrafascicular nerve electrode. AB - Artificial organs could be controlled using autonomic neural signals, because they exhibit rapid responses to physical needs similar to those of natural organs. A nerve electrode must satisfy many requirements to measure autonomous neural signals such as a long lifetime, high signal-to-noise ratio, multichannel recording, simple installation into a nerve fascicle, and good manufacturing productivity. The purpose of our study is to propose and evaluate a novel nerve electrode that satisfies these conditions, which to date has not been developed. A novel intrafascicular nerve electrode was designed, fabricated, and evaluated on autonomic nerves. Conventional extrafascicular and intrafascicular nerve electrodes were fabricated and tested for comparison to our novel intrafascicular nerve electrode. The novel intrafascicular nerve electrode had a 3-week lifetime, whereas the conventional extrafascicular nerve electrode had a 2-week lifetime. The signal-to-noise ratio was improved from 1.6 to 2.0 compared with the conventional extrafascicular nerve electrode. The novel intrafascicular nerve electrode was easier to install into a nerve fascicle and had better manufacturing productivity than the conventional intrafascicular nerve electrode. We succeeded in demonstrating the feasibility of our novel intrafascicular nerve electrode. PMID- 16340353 TI - First experimental evaluation of cardiac apex rotation with an epicardial coriolis force sensor. AB - Cardiac apex rotation, quantified by sophisticated techniques (radiopaque markers and tagged magnetic resonance), has been shown to provide a sensitive index of left ventricle (LV) dynamics. The authors describe the first experimental assessment of cardiac apex rotation using a gyroscopic sensor based on Coriolis force, epicardially glued on the apex. Dynamics of apex rotation were evaluated in a sheep at baseline, after a positive inotropic drug infusion, and after impairment of cardiac function induced by coronary ligation. To evaluate the efficacy of the sensor to monitor cardiac function, results were compared to contractility variations expressed by the maximum value of the first derivative of LV pressure (LVdP/dtMAX). After inotropic drug infusion, a parallel increasing trend resulted for LVdP/dtMAX, for the maximum value of angular velocity measured by the sensor, and for apex rotation angle derived from velocity signal (+146%, +155%, and +11% from baseline, respectively), whereas a decreasing trend of all three parameters resulted after coronary ligation (-35%, -31%, and -65%). The twist pattern also was altered from baseline. These initial results suggest that the use of an implantable rotation sensor based on Coriolis force can be an efficient and effective tool to assess LV torsional deformation both in normal and failing hearts. PMID- 16340354 TI - Computational design and experimental testing of a novel axial flow LVAD. AB - Thousands of cardiac failure patients per year in the United States could benefit from long-term mechanical circulatory support as destination therapy. To provide an improvement over currently available devices, we have designed a fully implantable axial-flow ventricular assist device with a magnetically levitated impeller (LEV-VAD). In contrast to currently available devices, the LEV-VAD has an unobstructed blood flow path and no secondary flow regions, generating substantially less retrograde and stagnant flow. The pump design included the extensive use of conventional pump design equations and computational fluid dynamics (CFD) modeling for predicting pressure-flow curves, hydraulic efficiencies, scalar fluid stress levels, exposure times to such stress, and axial fluid forces exerted on the impeller for the suspension design. Flow performance testing was completed on a plastic prototype of the LEV-VAD for comparison with the CFD predictions. Animal fit trials were completed to determine optimum pump location and cannulae configuration for future acute and long-term animal implantations, providing additional insight into the LEV-VAD configuration and implantability. Per the CFD results, the LEV-VAD produces 6 l/min and 100 mm Hg at a rotational speed of approximately 6300 rpm for steady flow conditions. The pressure-flow performance predictions demonstrated the VAD's ability to deliver adequate flow over physiologic pressures for reasonable rotational speeds with best efficiency points ranging from 25% to 30%. The CFD numerical estimations generally agree within 10% of the experimental measurements over the entire range of rotational speeds tested. Animal fit trials revealed that the LEV-VAD's size and configuration were adequate, requiring no alterations to cannulae configurations for future animal testing. These acceptable performance results for LEV-VAD design support proceeding with manufacturing of a prototype for extensive mock loop and initial acute animal testing. PMID- 16340355 TI - Hemodynamic modes of ventricular assist with a rotary blood pump: continuous, pulsatile, and failure. AB - Pulsatile operation of rotary blood pumps (RBPs) has received interest due to potential concern with nonphysiological hemodynamics. This study aimed to gain insight to the effects of various RBP modes on the heart-device interaction. A Deltastream diagonal pump (Medos Medizintechnik GmbH) was inserted in a cardiovascular simulator with apical-to-ascending aorta cannulation. The pump was run in continuous mode with incrementally increasing rotating speed (0-5000 rpm). This was repeated for three heart rates (50-100-150 bpm) and three levels of left ventricular (LV) contractility. Subsequently, the Deltastream was run in pulsatile mode to elucidate the effect of (de)synchronization between heart and pump. LV volume and pressure, arterial pressure, flows, and energetic parameters were used to evaluate the interaction. Pump failure (0 rpm) resulted in aortic pressure drops (17-46 mm Hg) from baseline. In continuous mode, pump flow compensated by diminished aortic flow, thus yielding constant total flow. High continuous rotating speed resulted in acute hypertension (mean aortic pressure up to 178 mm Hg). In pulsatile mode, unmatched heart and pulsatile pump rates yielded unphysiologic pressure and flow patterns and LV unloading was found to be highly dependent on synchronization phase. Optimal unloading was achieved when the minimum rotating speed occurred at end-systole. We conclude that, in continuous mode, a perfusion benefit can only be achieved if the continuous pump flow exceeds the preimplant (baseline) cardiac output. Pulsatile mode of support results in complex pressure and volume variations and requires accurate triggering to achieve optimal unloading. PMID- 16340356 TI - Use of a covered stent modification to produce a transcatheter valve: laboratory and animal testing. AB - Stent-based transcatheter valves continue to require large sheaths inappropriate for deployment in children. Low-profile covered stent valves (CSVs) were constructed by removing triangular sections from two sides of partially expanded Palmaz P308 stents before covering the stents with 0.1 mm polytetrafluoroethylene. Valves were carefully crimped onto balloon catheters and deployed in a pulsatile flow loop. With fixed afterload and pump output, flow, degree of stenosis, effect on pulse pressure, and ease of deployment were determined for each valve. In vivo transcatheter feasibility studies were then performed by disabling the aortic valve of two 25-kg pigs, and deploying transcatheter CSVs into their descending aorta. All transcatheter valves deployed consistently via sheaths three French sizes larger than the recommended sheath for their balloon and none created significant obstruction. With the bicuspid and supravalvar CSVs, the flow was 64% and 79% (respectively) of a commercially available valve. Angiograms revealed excellent acute CSV function after deployment with only mild regurgitation and without significant obstruction to systolic flow. Although long-term testing is required, a modified CSV design may have utility in low-profile pediatric transcatheter valve replacement. PMID- 16340357 TI - Hybrid small-caliber vascular prosthesis for coronary artery bypass grafting: a preliminary study of plasmin-treated fibrin-coated vascular prosthesis. AB - The potential use of plasmin-treated fibrin-coated vascular prosthesis (PF-V) for coronary artery bypass grafting (CABG) in animal models was investigated. PF-V grafts, 3 mm in internal diameter, were studied on 5 sheep in off-pump CABG model and on 18 rabbits in abdominal aortic bypass grafting (AABG) model. Patency, blood flow, angiography, Indium-111 platelet scintigraphy, and histology of the graft were evaluated. In the sheep CABG model, the PF-V grafts were patent for a range of 12 to 22 days without postoperative antiplatelet therapy. Graft flows ranged 58 to 90 ml/min until the day before graft occlusion by thrombus. In rabbit AABG model, the fibrin coating of the PF-V grafts was completely absorbed and replaced with neofibrin net between 7 and 14 days after implantation. Platelet depositions on the graft between 7 and 14 days after implantation were significantly higher than those at other periods (p < 0.05). The small-caliber PF V graft in sheep CABG model had a good blood flow with high antithrombogenicity in acute phase, but occluded over 2 to 3 weeks without antiplatelet agents after implantation. The current problem of the PF-V graft was a thrombus formation at the time of the degradation of fibrin coating. Further improvements are needed. PMID- 16340358 TI - Development of a small implantable right ventricular assist device. AB - The purpose of this program is to design, develop, and clinically evaluate a new, implantable right ventricular assist device (RVAD) that can be used as a component of an implantable biventricular assist device for patients with severe biventricular heart failure. The initial phase of this program resulted in a prototype RVAD, named DexAide, a modified version of the CorAide left ventricular assist device. In vitro testing was performed in a stand-alone circuit and in a true RVAD mode to evaluate pump performance. Pump flow and power were measured under various afterload and pump speed conditions. The pump performance requirements of 2 to 6 l/min and a pressure rise of 20 to 60 mm Hg were successfully met with pump speeds between 1,800 and 3,200 rpm. The nominal design point of 4 l/min and 40 mm Hg pressure rise was achieved at 2,450 +/- 70 rpm with a power consumption of 3.0 +/- 0.2 W. The initial in vitro testing met the design criteria for the new DexAide RVAD. Initial in vivo testing is under way, which will be followed by preclinical readiness testing and a pilot clinical trial in this 5-year program. PMID- 16340359 TI - Use of aortic valvo-pumps placed in valve annulus for long-term left ventricular assist. AB - To provide better anatomical fit and physiologic adaptation, three aortic valvo pumps with different dimensions were developed. Each pump has a rotor with an impeller and drive magnets and a stator consisting of a motor coil with iron core and an outflow guide vane. The devices had outer diameters of 21 mm, 23 mm, and 25 mm, respectively, and weighted 27 g, 31 g, and 40 g, respectively. Laboratory testing demonstrated that the rotating speed for maintaining a diastolic pressure of 80 mm Hg at zero flow rate should be 17500 rpm, 15000 rpm, and 12500 rpm, respectively; the largest flow at these same speeds will be 5 l/min, 7 l/min, and 10 l/min, respectively, with approximately 50 mm Hg pressure head increase. Therefore, these three pumps may meet the hemodynamic requirements of patients with body weights of 40-60 kg, 60-80 kg, and 80-100 kg. The first in vivo trial exhibited that the 25 mm valvo-pump can be sewn onto the aortic valve annulus of a 80 kg body weight pig without harm to adjacent organ functions. The devices occupy no additional anatomic space and deliver the blood directly from ventricle to the aorta, thus producing less physiologic disturbance to the natural circulation. Neither connecting conduits nor bypass circuits are necessary, thus eliminating the most dangerous sites of thrombosis in traditional left ventricular assist devices. PMID- 16340360 TI - Initial in vivo evaluation of the DexAide right ventricular assist device. AB - Despite the increasing use of left ventricular assist devices for patients with end-stage congestive heart failure, no implantable, centrifugal right ventricular assist devices (RVADs) are available for those patients with significant right ventricular failure. The DexAide RVAD was developed to provide an implantable RVAD option to surgeons. The aim of this study was to evaluate pump performance in an acute in vivo model. The DexAide RVAD, developed as a modified CorAide left ventricular assist device, was implanted between the right ventricle and the pulmonary artery in four healthy calves. Pump speed was varied from 1800 rpm to 3600 rpm. RVAD performance was analyzed acutely at baseline and under conditions of low circulating volume, high contractility, high pulmonary arterial pressure, vasodilation, and low contractility. Pump flow was well maintained even under conditions of high pulmonary arterial pressure and vasodilation, with the exception of low circulating volume. Under all conditions, pulmonary arterial pressures were not affected by changing pump speed. The DexAide RVAD demonstrated acceptable hemodynamic characteristics for use as an implantable RVAD in the initial acute studies. Further studies are ongoing to examine the biocompatibility of the pump under chronic conditions. PMID- 16340361 TI - Optimal peritoneal dialysis catheter type and exit site location: an anthropometric analysis. AB - An anthropometric analysis of 200 adult patients was performed to provide better guidance in catheter selection and placement. Height, weight, various abdominal wall measurements, and gender effects were analyzed. Suitability of Tenckhoff catheters with straight and preformed bends in the intercuff segment was evaluated regarding ability to produce deep pelvic position of the catheter tip and ideal exit site location. Conflicts with belt line and with skin creases and folds were recorded. Results showed that abdominal wall measurements varied widely by height and weight. Swan neck catheters with a downwardly directed external limb and exit site were significantly better suited for females (62% versus 27%, p < 0.0001). Tenckhoff catheters with straight intercuff segments with a laterally directed tunnel tract and exit site were significantly better matched to males (78% versus 30%, p < 0.0001). Neither catheter was suitable in 25% of subjects, emphasizing the need for an extended catheter system capable of remotely locating the exit site to the upper abdomen or chest without compromising pelvic position of the catheter tip. Appropriate preoperative evaluation with selection of the best suited catheter should replace the substandard practice of using a pet catheter to fit all patients and rigidly placing the insertion incision at a set location irrespective of body habitus. PMID- 16340362 TI - Phosphate clearance for bleach reprocessed polysulfone hemodialyzers: effects of electrostatic interactions. AB - Hyperphosphatemia is a major problem for hemodialysis patients due to inadequate phosphate clearance during dialysis. The objective of this study was to understand the effects of bleach reprocessing on phosphate clearance. Data were obtained using Fresenius polysulfone dialyzers exposed to bleach in an in vitro dialysis. The phosphate clearance decreased slightly after a single reprocessing cycle, with more than a 10% reduction observed after 15 cycles. Streaming potential measurements showed that bleach reprocessing caused an increase in the negative charge on the polysulfone membrane, suggesting that the change in phosphate clearance was due to an increase in electrostatic repulsion. The changes in clearance and surface charge were consistent with theoretical calculations of the effects of electrostatic interactions on solute diffusion. These results provide the first evidence that bleach reprocessing can reduce phosphate clearance due to electrostatic interactions between the phosphate anion and negatively charged dialysis membranes. PMID- 16340363 TI - Predicting dialysate sodium composition in sorbent dialysis using single point and multiple-dilution conductivity measurement. AB - This research establishes the ability to predict the sodium composition in dialysate from a single conductivity measurement over the wide range of concentrations of chloride, bicarbonate, and acetate that occur during sorbent dialysis. The ranges explored in mEq/L were sodium 100-180, chloride 76-143, bicarbonate 16-31, and acetate 4-11. Through mathematical optimization using a pattern search method, a single point measurement technique was shown to predict the total sodium concentration within approximately +/- 4.2 mEq/L in solutions with varying relative concentrations of chloride, bicarbonate, and acetate. The data analysis showed that the total sodium concentration can be predicted within +/- 2.1 mEq/L in most cases. Another potential approach to determining sodium concentration, a multiple-dilution measurement method, was tested and is also described. It is based on the varying relationship of activity to concentration for each of the sodium-anion pairs. This technique has practical limitations because of interactions between the various ions in solution at normal concentrations of dialysis along with the complexities involved in creating high dilutions of dialysate for on-line assays during dialysis. PMID- 16340364 TI - Prolonged stability of stored vancomycin, gentamicin, and heparin for use in the antibiotic-lock technique. AB - The antibiotic-lock technique has been effective in salvaging tunneled catheters in hemodialysis patients with bacteremia. However, a practical concern exists with respect to the stability of the antibiotics and heparin in normal saline, when stored for a prolonged period. Vancomycin, gentamicin, and heparin were diluted in normal saline to a final concentration of 100 microg/ml of each antibiotic and 5000 units/ml heparin. Fresh samples, and samples refrigerated at 4 degrees C for 48 hours, 1 week, 2 weeks, 3 weeks, and 4 weeks, were assayed (in triplicate) for gentamicin and vancomycin concentration and bactericidal activity (Schlichter test) using methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa. An anti-Xa activity assay was used for monitoring heparin anticoagulant activity of the fresh samples and samples refrigerated for 2 and 4 weeks. Mean (+/- SD) anti-Xa activity for heparin/vancomycin solution was 7900 +/ 173 u/ml, and for heparin/gentamicin solution was 7467 +/- 751u/ml; both were stable over a 4-week storage period. Mean bactericidal titer for vancomycin was 1:121 +/- 11, and for gentamicin was 1:242 +/- 22; both were stable over a 4-week storage period. Mean vancomycin concentration was 97 +/- 4 microg/ml, and gentamicin concentration was 86 +/- 3 microg/ml; both were stable over a 4-week storage period. Vancomycin and gentamicin in a heparin/saline solution can be stored at 4 degrees C for up to 4 weeks without adversely affecting antibiotic concentration, bactericidal activity, or heparin anticoagulant activity. PMID- 16340365 TI - Executive cognitive dysfunction without stroke after long-term mechanical circulatory support. AB - Among patients who receive heart transplantation (HTx) after long-term mechanical circulatory support (MCS), some show executive cognitive dysfunction without a history of stroke. Fifty HTx patients (19 patients on MCS for longer than 3 months before HTx and 31 patients without MCS as control group) were enrolled in the study. All subjects were men aged between 20 and 59 years without a history of stroke. Patients with MCS were divided into two groups: the AH-Thr group (n = 11), in which thrombus was detected in the left ventricular assist device (LVAD) and quickly removed (mean 3.3 times); and the AH group (n = 8), in which there was no detectable thrombus in the LVAD. The Trail Making Test (TMT) and the Wisconsin Card Sorting Test (WCST) were administered. The AH-Thr group showed poorer cognitive performance both in the TMT part B, with longer completion time (p < 0.05 versus the other two groups), and in the WCST, with more perseverative errors (p < 0.001 versus the other two groups). These data indicate that patients in the AH-Thr group showed executive cognitive dysfunction in set-shifting ability, suggesting frontal lobe damage. The conditions that facilitate thrombus formation in the LVAD may induce executive cognitive dysfunction without stroke. PMID- 16340366 TI - Effect of artificial lung compliance on right ventricular load. AB - Artificial lungs may serve as transplant alternatives to patients with respiratory failure. To provide a pulmonary replacement, the device cannot significantly alter right ventricular afterload. We sought to characterize cardiac load using a prototype artificial lung as a pulmonary replacement. An artificial lung was implanted into 13 adult sheep, 7 using a noncompliant original prototype and 6 using a modified device with a compliance chamber. Inflow was from the pulmonary artery and outflow into the left atrium. Pulmonary impedance was determined and pulse wave reflections analyzed. Resistance was similar in the original prototype artificial lung and in the native pulmonary circulation (3.0 vs 3.2 Woods units, p = 0.5). First harmonic impedance was significantly increased in the original prototype artificial lung (0.4 vs 5.9 Woods units, p < 0.01). High-amplitude pulse wave reflections were identified and right ventricular function was altered. The addition of compliance to the artificial lung reduced impedance at the first harmonic (5.9 vs 1.9 Woods units, p < 0.01), decreased the amplitude of pulse wave reflections, and normalized right ventricular function. A low-resistance but noncompliant artificial lung increases pulmonary impedance and alters right ventricular function. Addition of a compliance chamber reduces pulse wave reflections and normalizes ventricular function. PMID- 16340368 TI - Location of flow limitation in liquid-filled rabbit lungs. AB - The effects of end-inspiratory lung volume (EILV) and expiratory flow rate (Q) on the location of flow limitation in liquid-filled lungs were investigated by measuring pressure along the airways and by radiographic imaging. The lungs of New Zealand white rabbits were filled with perfluorocarbon to the randomly selected EILV of 20, 30, or 40 ml/kg, and the volume was actively drained at one of three Q: 2.5, 5.0, or 7.5 ml/s. The minimum pressures recorded by a movable catheter at locations along the airways show that flow limitation occurred in the main bronchi and trachea, and was independent of EILV and Q. The minimum pressure at the trachea was -80 mm Hg compared with values that were more positive than 10 mm Hg at a location 3 cm distal to the carina for all EILV and Q combinations. This location was confirmed by the lung images. The airway diameters gradually decreased with time, until flow limitation occurred. In airways distal to the collapse, there was not a significant decrease in diameter. Based on these data, we conclude that flow limitation in liquid-filled lungs occurs in the trachea and main bronchi and its location is independent of EILV or Q. PMID- 16340367 TI - Evaluation of a pumping assist lung that uses a rotating fiber bundle. AB - A paracorporeal respiratory assist lung (PRAL) is being developed for supplemental gas exchange to allow the native lungs of acute lung failure patients to heal. The device consists of a rotating annular microporous hollow fiber membrane bundle. The rotation augments the gas exchange efficiency of the device at constant flow-rate thereby uncoupling gas exchange and flow rate. The rotating fibers also enable the PRAL to pump the blood without the need for an additional pump or arterial cannulation. Blood flow rates will be between 500 and 750 ml/min with CO(2) removal rates of 100-130 ml/min. A prototype was manufactured with an overall surface area of 0.25 m. When rotated at 1500 rpm, CO(2) removal increased by 133% and O(2) transfer increased by 157% during an in vitro bovine blood study. The pumping of the rotating fiber bundle was assessed in a glycerol/water solution. At 1500 rpm, the PRAL generated 750 ml/min against 52 mm Hg pressure. Hemolysis of the device was assessed using in vitro bovine blood from a slaughterhouse. Plasma free hemoglobin levels were similar regardless of whether the rotating fibers were present in the PRAL, indicating that a rotating fiber bundle can be used to increase gas exchange without causing blood trauma. PMID- 16340369 TI - In vitro fluid mechanical effects of thoracic artificial lung compliance. AB - This in vitro study sought to determine what compliance minimizes thoracic artificial lung impedance and pump power output. A pulsatile pump drove 3.0 cP glycerol through a circuit consisting of an MC3 Biolung preceded by a piston cylinder (PC, n = 5) chamber with a variable compliance or a polyurethane (n = 4) chamber with a fixed, yet pressure-dependent, compliance. Each chamber was tested at flow rates of 1.8, 3.0, and 5.0 l/min and heart rates of 60, 75, and 100 bpm. Compliances, C, from 0-20 ml/mm Hg were tested in the PC chamber. Instantaneous pump outlet flow and pressure were acquired for determination of device zeroth and first harmonic input impedance, Z(0) and Z(1), and pump steady and pulsatile output powers, P(s) and P(p). PC chamber results indicate that Z(0), Z(1), P(s), and Pp were minimized at C > 1, 5, 0.5, and 4 ml/mm Hg, respectively. This suggests that C should be 1 ml/mm Hg at minimum and ideally 5 ml/mm Hg. The polyurethane chamber was statistically similar to the PC chamber at C = 1 ml/mm Hg when comparing Z(0) and P(s), but was statistically inferior when comparing Z(1) and P(p). The polyurethane compliance chamber, therefore, should be redesigned with greater compliance. PMID- 16340370 TI - Expiratory flow limitation during gravitational drainage of perfluorocarbons from liquid-filled lungs. AB - Flow limitation during pressure-driven expiration in liquid-filled lungs was examined in intact, euthanized New Zealand white rabbits. The aim of this study was to further characterize expiratory flow limitation during gravitational drainage of perfluorocarbon liquids from the lungs, and to study the effect of perfluorocarbon type and negative mouth pressure on this phenomenon. Four different perfluorocarbons (PP4, perfluorodecalin, perfluoro-octyl-bromide, and FC-77) were used to examine the effects of density and kinematic viscosity on volume recovered and maximum expiratory flow. It was demonstrated that flow limitation occurs during gravitational drainage when the airway pressure is < or = -15 cm H(2)O, and that this critical value of pressure did not depend on mouth pressure or perfluorocarbon type. The perfluorocarbon properties affect the volume recovered, maximum expiratory flow, and the time to drain, with the most viscous perfluorocarbon (perfluorodecalin) taking the longest time to drain and resulting in lowest maximum expiratory flow. Perfluoro-octyl-bromide resulted in the highest recovered volume. The findings of this study are relevant to the selection of perfluorocarbons to reduce the occurrence of flow limitation and provide adequate minute ventilation during total liquid ventilation. PMID- 16340371 TI - Effect of blood temperature on the efficacy of systemic leucodepletion during cardiopulmonary bypass: a prospective randomized clinical study. AB - The authors examined the effect of blood temperature within the cardiopulmonary bypass (CPB) circuit on the efficacy of an arterial line filter. Eighty patients undergoing elective, primary coronary artery bypass grafting under CPB were prospectively randomized in two equal groups. Blood temperature was kept at 35 degrees C in the first group and reduced to 28 degrees C in the second group. Twenty patients in each group had an arterial line LG6 filter attached onto CPB circuit. The other 20 patients in each group (controls) had a non-leukocyte depleting filter. Blood samples (10 ml) were taken before CPB, at 5 minutes on CPB, at 30 minutes on CPB, 5 minutes after aortic clamp removal, and 6 hours postoperatively. Leucocytes were counted under light microscopy. Activated leucocytes were identified using nitroblue tetrazolium staining. Patients undergoing leucodepletion had significantly lower total and activated leukocyte counts than control patients in both groups (p < 0.05). Patients having a leukocyte-depleting filter at a CPB temperature of 35 degrees C had significantly lower total leukocyte counts (p < 0.05) than those having a leukocyte-depleting filter at a CPB temperature of 28 degrees C (p < 0.05). However, there were not statistically significant differences in the activated leukocyte counts between the two leucodepleted groups (p > 0.05). This study shows that warm blood temperature within the CPB circuit has a positive effect on the overall leucodepleting efficacy of the LG6 filter. Activated leucocytes, however, seem to be depleted at similar rates irrespective of the blood temperature in the CPB circuit. PMID- 16340372 TI - Influence of volume replacement with colloids versus crystalloids in neonates on venoarterial extracorporeal membrane oxygenation on fluid retention, fluid balance, and ECMO runtime. AB - In this retrospective study, we compared the effects of colloid versus crystalloid fluid replacement on the clinical signs of capillary leakage syndrome in 30 neonates with pulmonary hypertension due to meconium aspiration syndrome on venoarterial membrane oxygenation (VA-ECMO). Before 2000, 15 neonates received volume replacement with a pasteurized plasma protein solution (3.8% albumin); after 2000, 15 neonates received normal saline. Patient characteristics and pre ECMO values did not differ between the two groups. Total fluid balance was also equal. Diuretic use was significantly higher in the colloid group (p < 0.001). The chest wall soft-tissue index was significantly higher in the crystalloid group (p < 0.005), as were the ventilator settings at the end of the ECMO runtime (p < 0.05). Serum colloid osmotic pressure, albumin, urea nitrogen, and creatinine were significantly higher in the colloid group (p < 0.0001, < 0.0001, < 0.001, and < 0.05, respectively). Duration of VA-ECMO, of artificial ventilation after ECMO treatment, and the mortality rate did not differ between the two groups. We conclude that volume replacement with crystalloids in neonates on VA-ECMO aggravated the edema in a preexisting situation of capillary leakage syndrome, whereas volume replacement with colloids could impair the kidney function. PMID- 16340373 TI - Practice patterns in neonatal cardiopulmonary bypass. PMID- 16340374 TI - Freestyle stentless bioprosthesis for active aortic infectious endocarditis with undetermined causative organism. AB - A 52-year-old woman developed congestive heart failure and was diagnosed with aortic regurgitation with approximately 20 mm of vegetation located on the noncoronary cusp of the aortic valve. The patient had undergone tooth extraction 3 months earlier, and various antibiotics had been given in the treatment of refractory high fever for 1 month. The damage to the aortic valve extended to the fibrous trigone near the mitral valve, and aggressive debridement of necrotic and infected tissue was done. The excised defect was repaired and reinforced with a pericardial patch, and the Freestyle stentless bioprosthesis was implanted using the full-root technique. Although the causative organism was never isolated, recurrent endocarditis has not occurred during 3 years of follow-up. The Freestyle stentless bioprosthesis was useful in the management of active aortic infectious endocarditis with undetermined causative organism for the prevention of recurrent endocarditis in the early postoperative period. PMID- 16340375 TI - Heparin-induced thrombocytopenia complicating support by the Berlin Heart. AB - Since 1992, miniaturized pulsatile air-driven ventricular assist devices (VADs), "Berlin Heart," have been used at many institutions (36 cases in North America in 19 different institutions) for pediatric use. Heparin-induced thrombocytopenia (HIT II) is a significant complication rarely reported in the setting of adult VAD support; no similar report exists concerning pediatric VAD support. We report on a 13-month-old, 8.1 kg girl who required LVAD support for cardiogenic shock of unclear etiology. The patient had a history of multiple surgical repairs for correction of complex congenital heart disease consisting of a series of left heart obstructive lesions (Shone's complex). Despite aggressive ventilatory and inotropic support, the patient continued to deteriorate and subsequently required extracorporeal life support. After 7 days of conventional venoarterial extracorporeal membrane oxygenation, a 10 ml Berlin Heart VAD was implanted. After implantation, the patient developed persistent low-grade fever of unclear etiology, gradual thrombocytopenia, and deterioration of renal function. On postimplant day 10, the pump required replacement because of concerns about an inlet valve thrombus; the explanted device demonstrated a nearly occlusive clot not appreciable from external inspection. Simultaneously, HIT II was diagnosed as a result of hematology workup for persistent thrombocytopenia. We discuss the unique challenges posed by HIT II complicating pediatric VAD support and in relation to the heparin coating of the device. PMID- 16340376 TI - Right heart bypass for acute traumatic respiratory distress syndrome. PMID- 16340377 TI - Epidural hemorrhage complicating extracorporeal life support in a neonate with respiratory failure. AB - Whereas bleeding represents the most common complication of a patient on extracorporeal life support, intracranial hemorrhage represents the most common bleeding complication. We report the first known case in the English literature of an epidural hemorrhage complicating extracorporeal life support in a neonate with respiratory failure. Ultrasound findings closely mimic those of a posterior fossa hemorrhage. We speculate that the coagulopathy associated with the use of ECLS may have contributed to the formation of the epidural hemorrhage. PMID- 16340378 TI - Effect of levodopa on healthy volunteers' facial emotion perception: an FMRI study. AB - OBJECTIVE: A link between the brain dopaminergic (DA) system and emotional processing seems to be supported by the DA nature of neural systems surrounding emotional recognition, the occurrence of emotional deficits in medical disorders involving a DA dysfunction, and the effect of DA agonists or antagonists on emotional processing. The authors tested the influence of levodopa administration on emotional processing in a functional MRI (fMRI) study of 10 elderly volunteers. METHODS: A placebo-controlled, cross-over experimental design was used. Subjects received either levodopa (100 mg) or placebo in 2 fMRI sessions. Performance was evaluated with a passive facial emotion perception test. RESULTS: During the placebo situation, the region-of-interest (ROI) analysis showed that emotional processing activated the bilateral amygdala. In levodopa volunteers, this activation was missing. The statistical comparison between the 2 situations (emotional vs control condition) revealed a highly significant reduction in activation of the bilateral amygdala for the levodopa fMRI session (P corrected <0.0001 in the left and P = 0.002 in the right amygdala). CONCLUSION: These results suggest that administration of levodopa to healthy volunteers directly or indirectly impairs the amygdalar activation during the emotional perception task. The authors hypothesized that amygdala activation may conform to an inverted U shaped function in relation to changing dopamine levels. PMID- 16340379 TI - Correlation between plasma homovanillic acid levels and the response to atypical antipsychotics in male patients with schizophrenia. AB - OBJECTIVE: The authors investigated the effects of atypical antipsychotic drugs olanzapine, perospirone, and quetiapine-on plasma homovanillic acid (pHVA) in male patients with chronic schizophrenia. METHODS: In this prospective, open label study, the subjects were 30 inpatients who were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, criteria for schizophrenia. The authors switched patients from typical antipsychotic drugs to olanzapine, perospirone, or quetiapine. Each patient gave informed consent for the research. pHVA was assessed before and after switching medications. RESULTS: After the switch, the authors found a significant improvement in psychotic symptoms, nonsignificant improvement in extrapyramidal symptoms, and a nonsignificant reduction in pHVA. In addition, the baseline pHVA correlated positively with the score changes from baseline in the Brief Psychiatric Rating Scale (BPRS) total, positive, and negative symptoms in the group with a whole sample and in the olanzapine-treated group, and with the score changes in the BPRS total and positive symptoms in the quetiapine-treated group. CONCLUSION: Our findings indicated that the preswitching pHVA levels could be used to predict changes in the psychotic symptoms of male patients with chronic schizophrenia when switching to atypical antipsychotic drugs. PMID- 16340380 TI - Hormonal profile effects following dehydroepiandrosterone (DHEA) administration to schizophrenic patients. AB - Dehydroepiandrosterone (DHEA) is an important neurosteroid and has demonstrated efficacy in the improvement of mood and energy. The authors previously reported the efficacy of DHEA augmentation in the management of negative, depressive, and anxiety symptoms of schizophrenia. To characterize further the effects of DHEA administration and to describe any hormonal effects following DHEA augmentation of antipsychotic medication, several hormones were measured (TSH, prolactin, testosterone, insulin, and estradiol) in 27 chronic schizophrenic inpatients receiving DHEA or placebo augmentation for 6 weeks. No significant changes in hormonal blood measures were noted. At the study end point, an association was noted between DHEA-S and TSH levels (P < 0.05) as well as a change in DHEA-S and insulin levels (P < 0.001). A modest nonsignificant decrease in plasma testosterone was observed following DHEA treatment. In subjects receiving DHEA, change in testosterone levels was associated with improvement in anxiety (P < 0.05) and illness severity (P < 0.05). Findings in this preliminary study demonstrate minimal effects on hormonal profiles despite the theoretic possibility that DHEA may elevate various hormones (eg, testosterone) with potential long-term adverse effects. DHEA's clinical efficacy appears to be mediated by other mechanisms, including direct effects on membrane channel coupled receptors. PMID- 16340381 TI - Venlafaxine and bladder function. AB - BACKGROUND: Occasional case reports describe urinary incontinence in patients taking the selective serotonin and norepinephrine reuptake inhibitor antidepressant venlafaxine. OBJECTIVE: In this study the authors investigated the possible effect of venlafaxine on urinary function in a series of 9 patients with urinary retention resulting from spinal cord lesions. They primarily sought to understand whether the reported venlafaxine-induced urinary incontinence was a specific drug-induced effect and, if so, whether venlafaxine might be an effective treatment of urinary retention. METHODS: During a 1-week baseline period, patients measured postvoiding residual volume through a catheter and recorded the number of micturitions within 24 hours. At the end of the baseline period, venlafaxine 75 mg extended-release on a once-daily evening administration schedule was added to their therapy for 1 week. RESULTS: None of the patients reported severe/uncontrollable side effects while taking venlafaxine. Extended release venlafaxine (75 mg/day) significantly reduced the postvoiding residual volume and increased the micturition rate; the volume diminished on the first day of treatment and remained stable over the ensuing days. CONCLUSION: These findings suggest that venlafaxine could be useful to improve voiding in patients with spinal cord disease. PMID- 16340382 TI - Levodopa intake increases plasma levels of S-adenosylmethionine in treated patients with Parkinson disease. AB - Metabolism of levodopa via the enzyme catechol-O-methyltransferase requires S adenosylmethionine (SAM) as a methyl donor. SAM caused Parkinson disease (PD) like symptoms in rodents. Therefore, SAM could contribute to a decreased efficacy of levodopa in the long term. SAM levels were significantly reduced in levodopa treated PD patients, but they showed increased enzyme methionine adenosyl transferase (MAT) activity, which induces SAM synthesis from methionine (MET). This may result from a rebound increase of SAM production. The objective of the study was to demonstrate an effect of acute levodopa intake on SAM synthesis in the plasma of treated PD patients. The authors measured SAM, MET, and levodopa plasma concentrations in 13 levodopa-treated PD patients before and after application of 125 mg levodopa/benserazide. Plasma levels of SAM and levodopa significantly increased, but MET concentrations did not significantly decrease. The SAM increase after levodopa intake may exert both a certain antidepressant and cognitive function improving effect. This is often observed in untreated PD patients who receive levodopa for the first time, or in more advanced, fluctuating PD patients, when they turn from the OFF to the ON phase. Because SAM in higher dosages may also counteract the antiparkinsonian efficacy of levodopa according to animal trials, this SAM increase may hypothetically contribute to the onset of wearing-off phenomena and other clinical signs of limited efficacy of levodopa during long-term treatment with levodopa in PD patients. PMID- 16340383 TI - Efficacy of topiramate and valproate in chronic migraine. AB - OBJECTIVES: The aim of this study was to compare the efficacy of sodium valproate and topiramate in treating chronic migraine. METHODS: Forty-nine patients with chronic migraine were randomly assigned to 1 of 2 groups of treatment: 750 mg/day valproate or 75 mg/day topiramate. Efficacy variables were number of days with headache over a 30-day period and changes in Migraine Disability Assessment (MIDAS) scores at 3 months. RESULTS: At baseline the 2 groups had similar numbers of days with headache and mean MIDAS scores. At the end of the treatment period, a significant reduction in 30-day headache frequency with respect to baseline (P < 0.00001) and a significant reduction in MIDAS scores (P < 0.00001) were recorded in both groups. There were no significant differences in beneficial effects between the 2 drugs. DISCUSSION: Valproate and topiramate seem to be able to manage successfully chronic migraine without substantial differences in efficacy and tolerability. This affords clear practical advantages-in the event of failure of or intolerance for one treatment, the patient may be switched to the other. PMID- 16340384 TI - Efficacy of levetiracetam in Huntington disease. AB - OBJECTIVE: Levetiracetam (LEV) is a novel antiepileptic drug characterized by a wide spectrum of action; no pharmacologic interaction and poor adverse events are reported. In animal models, effects of LEV are observed in basal ganglia. The aim of this study was to evaluate the efficacy of LEV in reducing involuntary movements in subjects affected by Huntington disease (HD). METHODS: This was a single-center, short-term, open-label, controlled study. Patients had LEV as add on therapy for 6 months. In the first visit patients were rated according to the Unified Huntington Disease Rating Scale. Every 2 months they were submitted to all these tests. LEV was added at the dose of 500 mg twice daily for the first 2 months and then the dosage was increased until 1000 mg twice daily for the next 4 more months. The authors enrolled 22 patients: 15 were assigned to the LEV group and 7 were enrolled as control subjects. RESULTS: No serious adverse events were experienced by the treated patients. After 6 months of treatment patients on LEV showed a significant reduction of involuntary movements, with a slight improvement of functional capacity compared with the control group. CONCLUSION: Results of this short-term, prospective, controlled study indicates that in HD patients, LEV is effective in reducing involuntary movements, thus improving the quality of life. PMID- 16340385 TI - A novel brain-targeted antioxidant (AD4) attenuates haloperidol-induced abnormal movement in rats: implications for tardive dyskinesia. AB - BACKGROUND: Tardive dyskinesia (TD), characterized by abnormal movements, is the major late-onset chronic side effect of antipsychotic treatment found in about 30% of those patients. The association of oxidative stress and the release of free radicals is one of the hallmarks of dopaminergic malfunctions and is one of the leading theories suggested for the pathophysiology of TD. To this day, no brain-targeted antioxidant has been tested as a potential treatment of TD. In light of this assumption, the authors chose a novel, low-molecular weight thiol antioxidant, N-acetyl cysteine amide (AD4), that crosses the blood-brain barrier as a possible treatment of TD. OBJECTIVE: To examine the protective effects of the novel brain-penetrating antioxidant AD4 on TD experimental models. METHODS: The typical vacuous chewing movement occurs in rats following chronic haloperidol injections (1.5 mg/kg/day intraperitoneally for 21 days). This purposeless mouth opening in the vertical plane is similar to TD symptoms in humans. The authors tested rats treated with haloperidol without or with AD4 in the drinking water (1 g/kg orally). Thiobarbituric acid reactive substances and anticarbonyl antibodies were used to measure oxidation of membranes and proteins. RESULTS: Haloperidol increased the vacuous chewing movements to 66.5 +/- 7.6 movements/5 minutes compared with 16.4 +/- 2.4 movements/5 minutes in untreated rats (P < 0.01). Coadministration of haloperidol and AD4 decreased the vacuous chewing movements level to 42.1 +/- 6.7 movements/5 minutes (P < 0.05). Haloperidol also increased the level of lipid peroxidation and protein oxidation in the rat brain, whereas coadministration with AD4 preserved their normal levels. CONCLUSION: Haloperidol causes behavioral abnormalities associated with oxidative stress in rats, similar to TD. AD4, the brain-targeted potent antioxidant, reduces the cellular oxidation markers and improves the typical clinical behavior. Hence, AD4 is a potential new treatment of antipsychotic-induced TD. PMID- 16340386 TI - Dystonic rabbit syndrome from citalopram. AB - OBJECTIVE: Two cases are described of dystonic rabbit syndrome induced by citalopram. This syndrome is a movement disorder with a 5-Hz rhythmic vertical motion of the mouth and lips without involvement of the tongue. METHOD: The patients were interviewed and examined, and additional history was taken from the medical records. The Naranjo adverse drug reaction rating scale was applied. Relevant literature was reviewed. RESULTS: Two patients developed dystonic rabbit syndrome soon after starting escitalopram 10 mg/day or citalopram 5 mg/day. Neither patient had any past or current exposure to a dopamine-blocking drug or any history of movement disorder. [Es]citalopram discontinuation led to disappearance of the movement disorder. The Naranjo scale indicates high probability of dystonic rabbit syndrome from citalopram. CONCLUSION: Citalopram can rapidly induce dystonic rabbit syndrome. This effect suggests that for some patients citalopram has neuropsychiatric effects similar to those of a dopamine blocking antipsychotic drug. This might be of concern with patients who cannot communicate well (eg, young children; patients with dementia, developmental disabilities, or aphasia). PMID- 16340387 TI - A case of acute thallotoxicosis successfully treated with double-filtration plasmapheresis. AB - The authors report a 48-year-old Chinese woman who presented with acute peripheral neuritis with progressive alopecia. Laboratory examinations disclosed a high blood concentration of thallium (97 microg/L) versus a normal value (0.9 microg/L), and she was diagnosed as having acute thallotoxicosis. After her hospitalization, the cutantest of dimercaptopropansulfonate sodium was positive and the patient refused to take Prussian blue because it caused constipation. She rapidly entered remission after assistance via double-filtration plasmapheresis (DFPP), suggesting the potential efficacy of DFPP for thallotoxicosis. PMID- 16340388 TI - Lithium intoxication associated with intramuscular ziprasidone in schizoaffective patients. AB - Antipsychotic agents have been used for the treatment of mood disorders in schizophrenic and schizoaffective patients. It has also been suggested that combinations of lithium and antipsychotics may be more effective than either class alone in treatment of schizoaffective patients. Lithium is known to interact with a variety of medications, including conventional and atypical antipsychotics. Although these combinations are generally well tolerated, they may sometimes lead to various adverse side effects. The authors report two schizoaffective patients with manic psychotic state and psychomotor agitation treated with lithium. Both patients developed symptoms of lithium intoxication after intramuscular ziprasidone treatment. To the best of their knowledge, this is the first description of such an adverse effect of this new atypical neuroleptic drug. They assume that lithium intoxication in the two presented cases was associated with the fast increase of ziprasidone serum level after its parenteral application, possibly affecting lithium excretion and thereby leading to lithium intoxication. PMID- 16340390 TI - Effects of Parkinson disease on two putative nondeclarative learning tasks: probabilistic classification and gambling. AB - OBJECTIVE: To assess performance on two nondeclarative (implicit) memory tasks of Parkinson disease (PD) patients without dementia in the earlier or later stages of the disease (Hoehn and Yahr Scale scores of 1-2.5 or 3-4, respectively). BACKGROUND: Different subtypes of nondeclarative memory appear to depend on different components of frontostriatal circuitry. Performance on a probabilistic classification learning (PCL) task was impaired by striatal damage (eg, in PD or Huntington disease) but not by circumscribed frontal lobe damage. On the other hand, performance on the Iowa Gambling Task (IGT) was impaired by damage to the prefrontal cortex. METHOD AND RESULTS: On the PCL, the learning of the control (age- and education-matched) group (n = 19) and the early PD group (n = 16) was comparable with each other, and both groups showed better performance than the later PD group (n = 16). On the IGT, the control group learned better than both of the PD groups. The control and early PD groups were similar on measures from the Wisconsin Card Sorting Test, Stroop Test, Mini-Mental State Examination, and Beck Depression Inventory II. CONCLUSIONS: The PCL and IGT tasks appear to rely on different parts of the frontostriatal circuitry in patients with early PD. The current finding that IGT performance was impaired in early PD implies ventromedial prefrontal cortical dysfunction early in the disease. PMID- 16340389 TI - Bupropion may improve restless legs syndrome: a report of three cases. AB - Restless legs syndrome (RLS) is a common disorder for which agents that enhance dopaminergic activity, including dopamine agonists and levodopa, are the treatment of choice. However, long-term use of dopaminergic drugs can cause unwanted effects such as rebound, tolerance, and augmentation. Bupropion, an inhibitor of dopamine and noradrenalin reuptake, is an antidepressant that modulates dopaminergic systems. The authors report that a low dose of bupropion rapidly and completely ameliorated RLS symptoms in 3 depressed patients within a few days of the initiation of treatment. To their knowledge, this is the first report to show that bupropion may be an effective alternative for treating RLS. Consequently, bupropion may be useful for the treatment of patients with both depression and RLS. PMID- 16340391 TI - An investigation of moral judgement in frontotemporal dementia. AB - OBJECTIVE: To investigate the basis of disturbed moral judgment in patients with frontotemporal dementia (FTD). BACKGROUND: FTD is characterized by difficulty in modulating social behavior. Patients lack social propriety and may perform sociopathic acts. In addition, FTD patients often lack empathy for others. These findings suggest alterations in the nature of morality in patients with FTD. METHOD: We administered an inventory of moral knowledge and two moral dilemmas to 26 patients with the frontal variant of FTD, 26 patients with Alzheimer disease (AD), and 26 normal control subjects. The FTD patients met Consensus Criteria for FTD and had corroborative frontal abnormalities on functional neuroimaging. The FTD and AD patients were comparably impaired on dementia measures. RESULTS: All these groups showed the retention of knowledge for moral behavior and the ability to make "impersonal" moral judgments. In contrast, the FTD patients were impaired in their ability to make immediate, emotionally based moral judgments compared with the patients with AD and the normal control subjects. CONCLUSIONS: These findings are consistent with an attenuation of the automatic emotional identification with others that is part of the innate moral sense. Such a disturbance may result from neurodegenerative disease affecting the ventromedial frontal cortex. PMID- 16340392 TI - The ubiquity of memory retrieval deficits in patients with frontal-striatal dysfunction. AB - OBJECTIVE AND BACKGROUND: Previous studies have shown that patients with frontal striatal dysfunction demonstrate improved performance on tests of recognition memory relative to free recall memory, suggesting deficits in retrieval processes. Not all studies, however, have indicated that all patients with frontal-striatal dysfunction display this profile. In this study, we examined the ubiquity of this "retrieval deficit" profile in a relatively large sample of patients with Parkinson disease (PD) or Huntington disease (HD). METHODS: Participants included 150 patients with PD and 65 patients with HD. Patients were classified as demonstrating a retrieval deficit or not based on a comparison of their standardized performances on the Recognition Discriminability and Long Delay Free Recall indices from the California Verbal Learning Test. RESULTS: Results indicated that 1) a retrieval deficit was more prevalent in patients with HD than PD, 2) this group difference emerged only in patients with at least a mild level of global cognitive impairment, and 3) even when the profile did emerge more frequently in patients with HD, it was present in only 44% of the patients. CONCLUSIONS: These findings suggest that not all patient groups with frontal-striatal dysfunction display a retrieval deficit profile, but in groups that do (ie, patients with HD), it is more likely to appear in individuals with greater cognitive impairment. PMID- 16340393 TI - Bradyphrenia and bradykinesia both contribute to altered speech in schizophrenia: a quantitative acoustic study. AB - OBJECTIVE: To evaluate the relative contributions of motor and cognitive symptoms on speech output in persons with schizophrenia (SZ). BACKGROUND: Studies of speech production in SZ suggest that atypical prosody (eg, pause) is related to clinical symptoms manifest in flat affect and alogia. Others have suggested that a more general motor slowing, bradykinesia, leads to measurable speech changes. METHOD: Thirteen participants with SZ and age-matched control subjects were included for between-group and by-task comparisons. Two levels of task complexity were analyzed acoustically to determine distinct and overlapping features of speech pause. RESULTS: For the free-speech task, group differences were found on measures of average pause duration, pause variability, percent pause, and cumulative pause time. Conversely, for the rote-speech task, group differences were found only on measures of average pause duration and pause variability. CONCLUSIONS: In persons with SZ, differences in the average and variability of pause duration may be reflected in speech motor slowing, whereas more global measures (eg, percentage pause) may better reflect a paucity of thought and idea generation related to the cognitive-linguistic aspects of free speech. These findings corroborate and extend the paucity of thought hypothesis in SZ to include an influence of motor slowing on speech production. PMID- 16340394 TI - Performance in Luria's fist-edge-palm test according to educational level. AB - OBJECTIVE: To investigate the performance of a large population with heterogeneous educational background in a test apparently uninfluenced by formal education. METHODS: We administered the Luria's fist-edge-palm test to 966 elderly individuals (299 illiterates) while screening for dementia in a Brazilian population. Individuals with scores in the Mini-Mental State Examination below the 25th percentile (adjusted for each of 4 educational levels) were considered suspect for dementia and excluded. The influence of educational level, age, and gender on the occurrence of failures and on the number of demonstrations for a successful reproduction was investigated. RESULTS: The final population comprised 745 individuals (238 illiterates). Forty-eight individuals (25 illiterates) were unable to reproduce the sequence. Logistic regression showed that illiteracy was associated with failure, whereas gender and age were not. The proportion of individuals failing to reproduce the sequence was inversely related to the years of schooling. For those individuals who were able to reproduce the sequence, the number of demonstrations for successful reproduction was also inversely related to the years of schooling. CONCLUSION: Illiteracy and low educational level are associated with impaired performance in the fist-edge-palm test, which may be the result of the use of different strategies according to educational level. PMID- 16340395 TI - Nature and development of Huntington disease in a nursing home population: The Behavior Observation Scale Huntington (BOSH). AB - OBJECTIVE: The Behavior Observation Scale Huntington (BOSH) was developed to obtain a detailed description of the manifestation of Huntington disease (HD) in the final stages. BACKGROUND: The Unified Huntington's Disease Rating Scale (UHDRS), developed to assess Huntington patients' clinical capacities, does not differentiate adequately in later stages of the disease. A scale easy to administer by nursing personnel for progression of the disease in later stages was needed. METHOD: Two pilot questionnaires preceded the final version of the BOSH. Observers administered the final version twice independently on 91 patients in 4 nursing homes. RESULTS: The BOSH contains 32 items in 3 subscales: 1) activities of daily living (ADL), 2) social-cognitive functioning, and 3) mental rigidity and aggression. Internal and interrater reliabilities were between 0.83 and 0.95. Rigidity and aggression become more frequent as the disease progresses; later on, this behavior diminishes. Social-cognitive capabilities deteriorate more rapidly in later stages. CONCLUSIONS: This study sought to develop a scale for assessing behaviors and individual differences in later stages of HD. The findings support the use of the BOSH in identifying these behaviors and differences. PMID- 16340396 TI - The effects of repetitive transcranial magnetic stimulation (rTMS) on procedural memory and dysphoric mood in patients with major depressive disorder. AB - OBJECTIVE: To study the effects of depression and treatment with repetitive transcranial magnetic stimulation (rTMS) on sequence learning. BACKGROUND: Prefrontal dysfunction in depression may affect sequence learning and be amenable to normalization by rTMS. METHOD: The serial reaction time test (SRTT) was administered to 19 patients with major depressive disorder (MDD) and 20 nondepressed control participants. MDD patients were examined before and following treatment with rTMS to the left dorsolateral prefrontal cortex in daily sessions of 1600 stimuli at 10 Hz and at an intensity of 110% of the motor threshold. Treatment occurred over a 2-week interval of time. RESULTS: MDD and nondepressed groups differed significantly with respect to baseline response speed. Following treatment with rTMS, MDD participants demonstrated significantly improved mood, improved response speed, and improved procedural learning. CONCLUSIONS: Findings suggest that rTMS over a 2-week period improves performance on tasks of response speed and procedural memory in patients with MDD. These cognitive effects are greater in those patients who showed a significant antidepressant effect to rTMS intervention. PMID- 16340397 TI - Diabetes and ischaemic heart disease: essential role for metabolic therapies. PMID- 16340398 TI - Rationale for a metabolic approach in diabetic coronary patients. AB - The incidence of ischaemic heart disease and acute myocardial infarction are greater in people with diabetes than in nondiabetic individuals. Heart disease patients with diabetes have a higher incidence of mortality during and following an acute myocardial infarction and a high risk for progression to heart failure post-infarction. The greater occurrence of ischaemic heart disease is partially due to a poorer coronary artery disease risk factor profile in diabetic patients, and, importantly, due to diabetes-induced abnormalities in the myocardium, termed 'diabetic cardiomyopathy'. The main metabolic abnormalities in the diabetic myocardium are impaired carbohydrate metabolism, specifically reduced pyruvate oxidation in the mitochondria and a greater reliance on fatty acids and ketone bodies as fuels. The healthy heart takes up glucose and lactate and converts them to pyruvate; however, in the diabetic heart there is a reduced capacity to oxidize pyruvate, and thus less glucose and lactate uptake. The defective metabolism is due to high circulating free fatty acids and ketone body concentrations in the plasma, resulting in greater acetyl-Co-enzyme A/Co-enzyme A and reduced nicotinamide adenonine dinucleotide/nicotinamide adenonine dinucleotide+ ratios in the mitochondria, and the subsequent inhibition of pyruvate dehydrogenase. Pharmacological inhibition of fatty acid oxidation during ischaemia increases myocardial pyruvate oxidation and provides clinical benefit to patients with stable angina or ischaemic left ventricular dysfunction. Recent clinical trials with trimetazidine, an inhibitor of the fatty acid beta-oxidation enzyme long chain 3-ketoacylthiolase, showed improvement in cardiac function and exercise performance in diabetic patients with ischaemic heart disease, illustrating the effectiveness of this approach in diabetes. PMID- 16340399 TI - Metabolic therapy for the diabetic patients with ischaemic heart disease. AB - Diabetic patients with ischaemic heart disease have a greater amount of myocardial ischaemia, often silent, and an increased incidence of heart failure compared to nondiabetic patients. This is the result of altered myocardial metabolism and accelerated atherogenesis with involvement of peripheral coronary segments causing chronic hypoperfusion and diffuse hybernation. In patients with diabetes mellitus and myocardial ischaemia, the metabolic changes occurring as a consequence of the mismatch between blood supply and cardiac metabolic requirements are heightened by the diabetic metabolic changes. An important metabolic alteration of diabetes is the increase in free fatty acid concentrations and increased muscular and myocardial free fatty acid uptake and oxidation. This increased uptake and utilization of free fatty acid during stress and ischaemia is responsible for the increased susceptibility of the diabetic heart to myocardial ischaemia and to a greater decrease of myocardial performance for a given amount of ischaemia compared to nondiabetic hearts. Given the metabolic alterations of the diabetic heart at rest and during episodes of myocardial ischaemia, a therapeutic approach aimed at an improvement of cardiac metabolism through manipulations of the utilization of metabolic substrates should result in an improvement of myocardial ischaemia and of left ventricular function. Modulation of myocardial free fatty acid metabolism should be the key target for metabolic interventions in patients with coronary artery disease with and without diabetes. In diabetic patients, the effects of modulation of free fatty acid metabolism should be even greater than those observed in patients without diabetes. The inhibition of FFA oxidation with trimetazidine improves cardiac metabolism at rest, decreases cardiac ischaemia and therefore prevents the decline of left ventricular function due to chronic hypoperfusion and repetitive episodes of myocardial ischaemia. Because of its effect on cardiac metabolism at rest, its effects on myocardial ischaemia and left ventricular function trimetazidine should always be considered for the treatment of diabetic patients with ischaemic heart disease with or without left ventricular dysfunction. PMID- 16340400 TI - Meeting the challenge of chronic ischaemic heart disease with trimetazidine. AB - The risk of cardiovascular events in patients with diabetes is similar to that of patients with previous history of ischaemic heart disease and the prevalence of ischaemic heart disease complicating diabetic syndromes is growing rapidly. Management of ischaemic heart disease in diabetic patients remains a challenge. Therapeutic options include several antianginal agents, and mechanical removal of coronary obstructions by percutaneous transluminal coronary angioplasty, and aorto-coronary by-pass surgery. Unfortunately, all treatments are less effective in diabetic patients than in nondiabetic patients. Direct modulation of cardiac metabolic alterations associated with the diabetic syndrome appears as a promising alternative for the management of ischaemic heart disease in diabetic patients. Results obtained in anginal patients with the metabolic agent trimetazidine are consistent with this hypothesis. PMID- 16340402 TI - Epidemiology of the diabetic heart. AB - Diabetes mellitus is a worldwide epidemic. Cardiovascular disease remains the major cause of morbidity and mortality in people with diabetes. Studies have suggested that increased risk of cardiovascular disease is not restricted to type II or type I diabetes mellitus, but extends to prediabetic stages such as impaired fasting glucose, impaired glucose tolerance, metabolic syndrome, and obesity. Insulin resistance, impaired fasting glucose, impaired glucose tolerance, and diabetes mellitus form a continuous sequence of risk for cardiovascular disease. Therefore, cardiovascular disease mortality and morbidity within the diabetes epidemic grow into vast proportions. Evidence also exists that diabetic patients have a high prevalence of heart failure or impaired diastolic and systolic cardiac function subsequent to the combination of coronary artery disease, hypertension, and diabetic cardiomyopathy. In view of the proportions of this new epidemic, prevention of diabetes and its prediabetic states is likely to be the most effective strategy to prevent serious cardiovascular events. PMID- 16340403 TI - Molecular biology for today's practicing intensivist. PMID- 16340404 TI - Transcription. PMID- 16340405 TI - Translation. PMID- 16340406 TI - Posttranslational protein modifications. PMID- 16340407 TI - Signal transduction overview. PMID- 16340408 TI - Nuclear factor-kappaB. PMID- 16340409 TI - Mitogen-activated protein kinase (MAPK). PMID- 16340410 TI - Akt/protein kinase B. PMID- 16340411 TI - Hypoxia-inducible factor-1 (HIF-1). PMID- 16340412 TI - Flow cytometry. PMID- 16340413 TI - Polymerase chain reaction: a landmark in the history of gene technology. PMID- 16340414 TI - Enzyme-linked immunosorbent assays (ELISAs). PMID- 16340415 TI - X-ray crystallography and structural biology. PMID- 16340416 TI - RNAi. PMID- 16340417 TI - Proteomics. PMID- 16340418 TI - Microarrays. PMID- 16340419 TI - Human genotyping. PMID- 16340420 TI - Toll-like receptors. PMID- 16340421 TI - Interleukin-1beta. PMID- 16340422 TI - Interleukin-6. PMID- 16340423 TI - Interleukin-8. PMID- 16340424 TI - Biology of interleukin-10 and its regulatory roles in sepsis syndromes. PMID- 16340425 TI - High mobility group box 1 (HMGB1). PMID- 16340426 TI - Macrophage migration inhibitory factor. PMID- 16340427 TI - Transforming growth factor-beta in critical illness. PMID- 16340428 TI - Complement. PMID- 16340429 TI - Coagulation. PMID- 16340430 TI - Eicosanoids. PMID- 16340431 TI - Nitric oxide: a clinical primer. PMID- 16340432 TI - Carbon monoxide. PMID- 16340433 TI - Reactive oxygen species. PMID- 16340434 TI - Neutrophils in the pathogenesis of sepsis. PMID- 16340435 TI - Monocytes/macrophages and sepsis. PMID- 16340436 TI - T lymphocytes. PMID- 16340437 TI - B cells. PMID- 16340438 TI - Cellular and molecular biology for intensivists: a primer--endothelial cells. PMID- 16340439 TI - Epithelial cells. PMID- 16340440 TI - Platelets. PMID- 16340441 TI - Apoptosis. PMID- 16340442 TI - Mechanisms of cell necrosis. PMID- 16340443 TI - The role of gap junctions in health and disease. PMID- 16340444 TI - Metabolic failure. PMID- 16340446 TI - Prenatal ethanol exposure in rats decreases levels of complexin proteins in the frontal cortex. AB - BACKGROUND: Rodents that are prenatally exposed to ethanol have been shown to exhibit a wide range of cognitive deficits, including impairments in memory, attention and executive function. To determine a potential molecular substrate for cognitive dysfunction in adulthood, we measured regional levels of the presynaptic proteins complexin I and II in a rat model of prenatal ethanol exposure, as levels of these proteins are altered in cognitive-related synaptic plasticity. METHODS: Pregnant female rats received either a liquid ethanol diet (36% ethanol-derived calories) or a liquid control diet (maltose-dextrin isocalorically substituted for ethanol, matched in amount [g/kg body wt/day of gestation] to an ethanol-consuming partner), or were given ad libitum-fed access to standard laboratory chow and water. Levels of complexin I, II and the ubiquitous presynaptic marker synaptophysin were measured in the frontal cortex and hippocampus of adult male offspring, using ELISA. RESULTS: Prenatal exposure to ethanol did not alter levels of presynaptic proteins in the hippocampus or levels of synaptophysin in the prefrontal cortex. However, rats prenatally exposed to ethanol displayed significantly lower levels of both complexin I and II in the prefrontal cortex compared to control animals. CONCLUSIONS: These data indicate that prenatal exposure to ethanol is associated with a selective loss of complexin proteins in the frontal cortex. These proteins are known to be important for activity-dependent neurotransmission, and have previously been shown to mediate synaptic plasticity and cognition. These combined findings suggest that further study of complexin proteins as a substrate for cognitive impairment related to prenatal exposure to ethanol is warranted. PMID- 16340447 TI - A cross-generational comparison of alcohol challenges at about age 20 in 40 father-offspring pairs. AB - BACKGROUND: A low level of response (LR) to alcohol is one of several genetically influenced phenotypes associated with an elevated risk for heavy drinking and alcoholism. While most studies support the influence of genes for this characteristic, no data to date have addressed how LR established from alcohol challenges performs in similarly aged subjects across generations. METHODS: Between 1978 and 1988, 18-to-25-year-old non-alcohol-dependent Caucasian male drinkers participated in the San Diego Prospective Study alcohol challenges. The paradigms included self-reports of feelings of "High" and "Intoxication," as well as alcohol-related changes in body sway. In recent years, 40 18-to-29-year-old offspring of 25 of these original probands were tested using a similar protocol. RESULTS: Despite the passage of two decades between laboratory sessions across generations, for family history positive (FHP) subjects, significant positive correlations were observed for subjective feelings of intoxication and body sway after alcohol. Parent-offspring correlations were in the predicted direction for subjective feelings for family history negatives (FHNs), but were not significant. Across offspring, LR values were lower for FHPs overall, with significant differences at 60 or 90 min for five items. CONCLUSIONS: The similarities in LR across generations, while not proving heritability, are consistent with prior reports regarding genetic influences in the LR to alcohol. The significant correlations across generations and over two decades support the reliability of the alcohol challenge results. PMID- 16340448 TI - -238 G>A polymorphism of tumor necrosis factor alpha gene (TNFA) is associated with alcoholic liver cirrhosis in alcoholic Spanish men. AB - BACKGROUND: The tumor necrosis factor alpha gene (TNFA) has been recently associated to alcoholic steatohepatitis. We have analyzed the distribution of genotypes and alleles of two polymorphisms at positions -238 and -308 in the promoter region of the TNFA gene in a Spanish male population of alcoholics with and without alcoholic liver cirrhosis. METHODS: 149 male alcoholics (84 without alcoholic liver disease, and 65 with alcoholic liver cirrhosis) and 90 control subjects were included. Genotyping was done by polymerase chain reaction and digestion with restriction enzymes. RESULTS: No significant differences in the distribution of genotypes and alleles of the -308 TNFA gene polymorphism were observed between alcoholics and non-alcoholics, or between alcoholics with liver cirrhosis and those without liver disease. However, we found an association between the -238 TNFA polymorphism and alcoholic liver cirrhosis; the frequency of the heterozygous genotype being significantly higher in alcoholics with cirrhosis than in those without liver damage. CONCLUSION: The -238 TNFA-A allele is associated with a higher risk to develop alcoholic liver cirrhosis. This polymorphism could be considered as a genetic factors that confer predisposition to suffer liver cirrhosis in the alcoholic population of Castile and Leon. PMID- 16340449 TI - Chronic ethanol exposure stimulates endothelial cell nitric oxide production through PI-3 kinase-and hsp90-dependent mechanisms. AB - BACKGROUND: Chronic ethanol (EtOH) ingestion increases the incidence of the Acute Respiratory Distress Syndrome (ARDS), a severe form of acute lung injury characterized by endothelial and epithelial barrier dysfunction. The regulated production of nitric oxide (NO) by the endothelium plays a central role in normal vascular function, and alterations in NO production have been implicated in barrier dysfunction. Although previous reports examined the impact of acute EtOH stimulation on endothelial NO production, this study extends those observations to clarify mechanisms of chronic EtOH-mediated alterations in endothelial nitric oxide synthase (eNOS) expression and NO production. METHODS: Porcine pulmonary artery endothelial cells (PAEC) were treated with EtOH (0.04-0.16%, w/v) for 72 hr in sealed chambers to prevent evaporation. NO release and eNOS expression were determined to examine the effect of chronic EtOH stimulation on endothelial NO metabolism. RESULTS: While there was no change in the extent of phosphorylated eNOS at ser, chronic EtOH stimulation caused dose-dependent increases in NO production and increased eNOS expression, effects that were attenuated by the transcriptional inhibitor, alpha-amanitin (AA), and wortmannin, a specific phosphatidylinositol 3 kinase (PI3 K) inhibitor. EtOH stimulation also increased eNOS interaction with heat shock protein (hsp90), a molecular chaperone known to enhance eNOS activity. Geldanamycin, an hsp90 inhibitor, attenuated chronic EtOH mediated increases in NO production. CONCLUSIONS: These results indicate that chronic EtOH exposure increases endothelial NO production by increasing eNOS protein levels through PI3 K-dependent up regulation of eNOS gene transcription and by increasing interactions between eNOS and hsp90. These findings clarify mechanisms by which chronic EtOH stimulation modulates vascular endothelial function and suggest new targets for investigation and intervention in EtOH induced alterations in susceptibility to lung injury. PMID- 16340450 TI - Acute alcohol withdrawal is associated with c-Fos expression in the basal ganglia and associated circuitry: C57BL/6J and DBA/2J inbred mouse strain analyses. AB - BACKGROUND: The DBA/2J (D2) and C57BL/6J (B6) mouse strains are the most widely studied genetic models of severe and mild acute alcohol withdrawal, respectively. Previous studies have identified quantitative trait loci and genes involved in risk for acute ethanol withdrawal using mapping populations derived from the D2 and B6 strains, but the brain region(s) and circuit(s) by which these genes and their protein products influence ethanol physiological dependence and associated withdrawal remain to be elucidated. METHODS: B6 and D2 were administered a sedative-hypnotic dose of ethanol (4 g/kg) or saline (control) and returned to their home cages where they were left undisturbed for 7 hr, which has been shown in previous studies to correspond to peak acute ethanol withdrawal severity. The mice were then euthanized and assessed for their numbers of c-Fos immunoreactive neurons across 26 brain regions. The question addressed was whether or not ethanol-withdrawn D2 and B6 mice differed in c-Fos induction (neural activation) within circuitry that could explain the severe ethanol withdrawal of the D2 strain and the mild ethanol withdrawal in B6 strain mice. RESULTS: At peak acute ethanol-withdrawal ethanol-withdrawn D2 and B6 mice differed in neural activation within the basal ganglia, including the subthalamic nucleus and the two major output nuclei of the basal ganglia (the medial globus pallidus and the substantia nigra pars reticulata). Genotype-dependent c-Fos induction was also apparent in associated circuitry including the lateral septum, the ventral tegmental area, the nucleus accumbens core, the dorsolateral caudate putamen, the substantia nigra pars compacta, the cingulate and entorhinal cortices, and the ventral pallidum. D2 and B6 mice showed comparable neural activation in the bed nucleus of the stria terminalis, and the nucleus accumbens shell. CONCLUSIONS: The present studies are the first to use immediate early gene product expression to assess the pattern of neural activation associated with acute ethanol withdrawal. Our results point to the involvement of an extended basal ganglia circuit in genetically determined differences in acute ethanol withdrawal. Based on these data, we suggest that quantitative trait genes (QTGs) involved in acute ethanol withdrawal exert their effects on this phenotype via one or more of the brain regions and circuits identified. As more information becomes available that integrates neural circuit and QTG analyses, the precise mechanisms by which QTGs affect ethanol physiological dependence and associated withdrawal will become apparent. PMID- 16340451 TI - Hybrid C57BL/6J x FVB/NJ mice drink more alcohol than do C57BL/6J mice. AB - BACKGROUND: From several recent strain surveys (28 strains: Bachmanov et al., personal communication; 22 strains: Finn et al., unpublished), and from data in >100 other published studies of 24-hr two-bottle ethanol preference, it is known that male C57BL/6 (B6) mice self-administer about 10-14 g/kg/day and that female B6 mice self-administer about 12-18 g/kg/day. No strain has been found to consume more ethanol than B6. In one of our laboratories (Texas), we noted a markedly greater intake of ethanol in an F1 hybrid of B6 and FVB/NJ (FVB) mice. METHODS: To confirm and extend this finding, we repeated the study at another site (Portland) using concentrations up to 30% ethanol and also tested B6xFVB F1 mice in restricted access drinking procedures that produce high levels of alcohol intake. RESULTS: At both sites, we found that B6xFVB F1 mice self-administered high levels of ethanol during two-bottle preference tests (females averaging from 20 to 35 g/kg/day, males 7-25 g/kg/day, depending on concentration). F1 hybrids of both sexes drank significantly more 20% ethanol than both the B6 and FVB strains. Female F1 hybrids also drank more 30% ethanol. In the restricted access tests, ethanol consumption in the F1 hybrids was equivalent to that in B6 mice. CONCLUSIONS: These data show that this new genetic model has some significant advantages when compared to existing inbred strains, and could be used to explore the genetic basis of high ethanol drinking in mice. PMID- 16340452 TI - Aldehyde dehydrogenase 2 gene targeting mouse lacking enzyme activity shows high acetaldehyde level in blood, brain, and liver after ethanol gavages. AB - BACKGROUND: Previously, we created an aldehyde dehydrogenase 2 gene transgenic (Aldh2-/-) mouse as an aldehyde dehydrogenase (ALDH) 2 inactive human model and demonstrated low alcohol preference. In addition, after a free-choice drinking test, no difference in the acetaldehyde level was observed between the Aldh2-/- and wild type (Aldh2+/+) mice. The actual amounts of free-choice drinking were so low that it is uncertain whether these levels are pharmacologically and/or behaviorally relevant in either strain. To elucidate this uncertainty, we compared the ethanol and acetaldehyde concentration in the blood, brain, and liver between the Aldh2-/- and Aldh2+/+ mice after ethanol gavages at the same dose and time. METHOD: We measured differences in the ethanol and acetaldehyde levels between the Aldh2-/- and Aldh2+/+ mice by headspace gas chromatography mass spectrometry (GC-MS) after ethanol gavages at the same dose and time. RESULTS: Significantly higher blood acetaldehyde concentrations were found in the Aldh2-/- mice than in the Aldh2+/+ mice 1 hr after the administration of ethanol gavages at doses of 0.5, 1.0, 2.0, and 5.0 g/kg. The blood acetaldehyde concentrations in the two strains were 2.4 vs. 0.5, 17.8 vs. 1.9, 108.3 vs. 4.3, and 247.2 vs. 14.0 (microM), respectively. In contrast, no significant difference was observed in the blood ethanol concentrations between the Aldh2+/+ and Aldh2-/ mice. The aldehyde dehydrogenase 2 enzyme metabolized 94% of the acetaldehyde produced from the ethanol as calculated from the area under the curve (AUC) of acetaldehyde when ethanol was administered at a dose of 5.0 g/kg. CONCLUSIONS: These data indicate that mouse ALDH2 is a major enzyme for acetaldehyde metabolism, and the Aldh2-/- mice have significantly high acetaldehyde levels after ethanol gavages. PMID- 16340453 TI - Differences in the peripheral levels of beta-endorphin in response to alcohol and stress as a function of alcohol dependence and family history of alcoholism. AB - BACKGROUND: Evidence indicates that both genetic and environmental factors, such as stress, may play an important role for the development of alcoholism, while beta-endorphin may be implicated in the control of alcohol consumption. The objective of the present studies was to test the hypothesis that there are differences in the response of the pituitary beta-endorphin system to stress as a function of family history of alcoholism and alcohol dependence. METHODS: The response of the pituitary beta-endorphin to a placebo or an alcohol (0.50 g ethanol/kg) drink and to a stress task performed 30 min following ingestion of either the placebo or the alcohol drink was measured in social and heavy drinkers with [high risk (HR)] and without [low risk (LR)] a family history of alcoholism. Thus, each subject participated in 4 experimental sessions given on different days in a randomized order. Four groups of subjects were investigated: 1) low risk nonalcoholics (LRNA); 2) high risk nonalcoholics (HRNA), 3) low risk alcoholics (LRA); and 4) high risk alcoholics (HRA). Plasma beta-endorphin was estimated prior to and for 3.5 hr post-stress. Changes in the concentration of plasma beta-endorphin following ingestion of either the placebo or alcohol drink without performance of the stress task served as controls to compare the stress induced changes. RESULTS: Basal plasma beta-endorphin levels were higher in LRNA than LRA, HRNA and HRA participants, while basal plasma beta-endorphin levels were higher in LRA than those in HRNA and HRA participants. Furthermore, there was no significant difference in the plasma beta-endorphin levels between HRNA and HRA participants. Stress, induced a significant increase in plasma beta endorphin concentration in all four groups of participants. However, the stress induced increase in plasma beta-endorphin levels was more pronounced in LRNA than HRNA, LRA and HRA participants. Thus, alcohol dependence decreased the basal plasma beta-endorphin levels in LR only, as well as the stress induced increase in plasma beta-endorphin levels of participants without, but not of those with, a family history of alcoholism. Alcohol prior to stress attenuated the stress induced increase in plasma beta-endorphin levels of all four groups of participants. CONCLUSIONS: The present data indicates that there are differences in both, the basal plasma beta-endorphin levels as well as the response of the pituitary beta-endorphin to stress as a function of family history of alcoholism and alcohol dependence. Thus, in HR individuals a dysfunction in the activity of the pituitary beta-endorphin system predates the development of alcoholism, while in LR individuals it develops following alcohol dependence. Furthermore, alcohol dependence did not alter the alcohol-induced attenuation of beta-endorphin response to stress. PMID- 16340455 TI - Alcoholics anonymous careers: patterns of AA involvement five years after treatment entry. AB - BACKGROUND: Most formal treatment programs recommend Alcoholics Anonymous (AA) attendance during treatment and as a form of aftercare, but we know very little about treatment seekers' patterns of AA involvement over time and how these relate to abstinence. METHOD: This paper applies latent class growth curve modeling to longitudinal data from 349 dependent drinkers recruited when they were entering treatment and were re-interviewed at one or more follow-up interviews one, three and five years later, and who reported having attended AA at least once. RESULTS: Four classes of AA "careers" of meeting attendance emerged: The low AA group mainly just attended AA during the 12 months following treatment entry. The medium and high AA groups were characterized by stable attendance at the second and third follow-ups-at about 60 meetings a year for the medium group and over 200 meetings per year for the high group, followed by slight increases for the medium group and slight decreases for the high group by year five. The declining AA group doubled its meeting attendance postbaseline, to almost 200 meetings during the year following treatment entry, but by year five they were only attending about six meetings on average. Decreases in AA meetings did not necessarily signal disengagement from AA; at the five-year follow-up, a third of the low AA group and over half of the declining AA group said they felt like a member of AA. Activities other than meeting attendance, such as having a sponsor, otherwise paralleled the meeting careers, but social networks were similar by year five. Rates of abstinence by year five (for the past 30 days) were 43% for the low AA group, 73% for the medium group, 79% for the high group and 61% for the declining group. Rates of dependence symptoms and social consequences of drinking did not differ between the groups at year five. CONCLUSIONS: The prototypical AA careers derived empirically are consistent with anecdotal data about AA meetings: some never connect; some connect but briefly; and others maintain stable (and sometimes quite high) rates of AA attendance. However, contrary to AA lore, many who connect only for a while do well afterwards. PMID- 16340454 TI - Autosomal linkage analysis for the level of response to alcohol. AB - BACKGROUND: The level of response (LR) to alcohol is a genetically-influenced phenotype related to the alcoholism risk. Usually measured by evaluating psychological and physiological changes that follow the administration of alcohol, the heritability of LR is estimated to be between 0.4 and 0.6, and efforts are being made to find genes related to this phenotype. This paper presents data from a family-based genome with linkage analysis focusing on alcohol challenge determinants of LR. METHODS: The subjects were 18-to-29-year old sibling pairs with at least one parent who was alcohol-dependent and who had experience with alcohol but were not yet alcohol-dependent themselves. Both members of the sibling pairs were given oral alcohol challenges (0.75-0.90 ml/kg of ethanol for females and males, respectively), with LR established using the Subjective High Assessment Scale (SHAS) and changes in body sway (BS) repeatedly over a 3.5-hr. period. Blood samples from siblings and at least one parent were genotyped using 811 microsatellite markers, with results evaluated using several related variance component approaches as implemented in SOLAR for continuous traits. In addition, association was tested using single nucleotide polymorphisms (SNPs) within the KCNMA1, HTR7 and SLC18A2 genes that may relate to a finding on chromosome 10. RESULTS: Data were generated from 238 sib-pairs representing 365 individuals (41.6% were males) from 165 families. The most consistent results across methods and samples were observed for SHAS on chromosome 10 between 120 and 140 cM (with a maximum LOD score of 2.6 at 122 cM), and a second region of possible interest at 173 cM (LOD = 1.2). Statistical analysis with the KCNMA1, HTR7 and SLC18A2 genes, which lie in the support region of interest revealed no evidence for association after correction for multiple comparisons. CONCLUSIONS: These evaluations from the largest known alcohol challenge-based genetic study to date highlight the potential importance of genes on chromosome 10 as possible contributors to the low LR to alcohol as a risk factor for alcoholism. PMID- 16340456 TI - Applying clinical significance methodology to alcoholism treatment trials: determining recovery outcome status with individual- and population-based measures. AB - BACKGROUND: The current analysis applies clinical significance methodology to alcoholism treatment outcome research using data available from Project MATCH. Because of its high internal validity and its inclusion of multiple measures assessing multiple outcome dimensions, MATCH was considered an ideal study to explore the utility of this methodology. METHODS: Data reported here are from a total of 1,726 participants enrolled in either aftercare (n = 774) or outpatient (n = 952) arms of the study. First, a cutoff score was determined differentiating functional versus dysfunctional status on three outcome measures: percent days abstinent (PDA), mean drinks per drinking day (DDD) and negative consequences of alcohol use. Second, the reliable change in pre- to post-treatment scores on these three measures was calculated. RESULTS: The results reported herein support the importance of distinguishing between statistical and clinical significance of outcomes. During three months post-treatment, approximately one-half of the treated patients were "recovered" (i.e., both functional and reliably changed) with respect to both PDA (i.e., 51%) and negative consequences of drinking (i.e., 47%); however, only about one-third of individuals remained recovered throughout the full one-year follow-up period (i.e., 33% on PDA and 35% on negative consequences). These individual-based change outcomes compared similarly to a population-based indicator of heavy drinking. Alternatively, only about one quarter of participants were recovered using two distinct criteria for mean DDD (i.e., 23-29%), and even fewer participants remained recovered on mean DDD over the full one-year follow-up period (i.e., about 14-18%). CONCLUSIONS: Based on study limitations, more work is required to make clinical significance methodology practically useful to alcoholism treatment trials including more precise definitions of functional status and relative change as well as better interpretation of the inter-relationship between multiple measures assessing multiple outcome domains. PMID- 16340457 TI - The Alcohol Use Disorders Identification Test (AUDIT) as a screening tool for excessive drinking in primary care: reliability and validity of a French version. AB - BACKGROUND: Excessive drinking is a major problem in Western countries. AUDIT (Alcohol Use Disorders Identification Test) is a 10-item questionnaire developed as a transcultural screening tool to detect excessive alcohol consumption and dependence in primary health care settings. OBJECTIVES: The aim of the study is to validate a French version of the Alcohol Use Disorders Identification Test (AUDIT). METHODS: We conducted a validation cross-sectional study in three French speaking areas (Paris, Geneva and Lausanne). We examined psychometric properties of AUDIT as its internal consistency, and its capacity to correctly diagnose alcohol abuse or dependence as defined by DSM-IV and to detect hazardous drinking (defined as alcohol intake >30 g pure ethanol per day for men and >20 g of pure ethanol per day for women). We calculated sensitivity, specificity, positive and negative predictive values and Receiver Operator Characteristic curves. Finally, we compared the ability of AUDIT to accurately detect "alcohol abuse/dependence" with that of CAGE and MAST. RESULTS: 1207 patients presenting to outpatient clinics (Switzerland, n = 580) or general practitioners' (France, n = 627) successively completed CAGE, MAST and AUDIT self-administered questionnaires, and were independently interviewed by a trained addiction specialist. AUDIT showed a good capacity to discriminate dependent patients (with AUDIT > or =13 for males, sensitivity 70.1%, specificity 95.2%, PPV 85.7%, NPV 94.7% and for females sensitivity 94.7%, specificity 98.2%, PPV 100%, NPV 99.8%); and hazardous drinkers (with AUDIT > or =7, for males sensitivity 83.5%, specificity 79.9%, PPV 55.0%, NPV 82.7% and with AUDIT > or =6 for females, sensitivity 81.2%, specificity 93.7%, PPV 64.0%, NPV 72.0%). AUDIT gives better results than MAST and CAGE for detecting "Alcohol abuse/dependence" as showed on the comparative ROC curves. CONCLUSIONS: The AUDIT questionnaire remains a good screening instrument for French-speaking primary care. PMID- 16340458 TI - Cost-benefit analysis of a new alcohol biomarker, carbohydrate deficient transferrin, in a chronic illness primary care sample. AB - BACKGROUND: Carbohydrate Deficient Transferrin (CDT) is a new alcohol biomarker recently approved by the Food and Drug Administration for alcohol screening. Limited information is available on the economic benefits of alcohol biomarkers. Our objective was to conduct a cost-benefit analysis (CBA) of the CDT test in a primary care sample of patients being treated for diabetes and hypertension. METHODS: A decision tree was created using data from national surveys, published literature, and two brief intervention trials conducted in primary care settings. The decision tree was used to estimate the costs and benefits of CDT under different scenarios. RESULTS: For the base case, utilizing CDT in addition to patient self-report resulted in an increase from 28 to 53 problem drinking cases identified out of 70 cases screened. With increased detection and subsequent intervention, the average medical and legal costs were far lower in the CDT arm of the study. When these avoided costs were incorporated into the model, an overall savings of $212.30 per patient was realized with CDT testing. Monte Carlo analysis also indicated a trend toward cost savings, with a mean cost savings of approximately $353 and a range of $1,619 in savings to $450 in costs for 1,000 simulations of the decision tree model. CONCLUSION: This CBA suggests that the addition of routine CDT screening to patient self-report may provide positive net economic benefits in primary care settings. PMID- 16340459 TI - A drink is a drink? Variation in the amount of alcohol contained in beer, wine and spirits drinks in a US methodological sample. AB - BACKGROUND: Empirically based estimates of the mean alcohol content of beer, wine and spirits drinks from a national sample of US drinkers are not currently available. METHODS: A sample of 310 drinkers from the 2000 National Alcohol Survey were re-contacted to participate in a telephone survey with specific questions about the drinks they consume. Subjects were instructed to prepare their usual drink of each beverage at home and to measure each alcoholic beverage and other ingredients with a provided beaker. Information on the brand or type of each beverage was used to specify the percentage of alcohol. RESULTS: The weighted mean alcohol content of respondents' drinks was 0.67 ounces overall, 0.56 ounces for beer, 0.66 ounces for wine and 0.89 ounces for spirits. Spirits and wine drink contents were particularly variable with many high-alcohol drinks observed. CONCLUSIONS: While the 0.6-ounce of alcohol drink standard appears to be a reasonable single standard, it cannot capture the substantial variation evident in this sample and it underestimates average wine and spirits ethanol content. Direct measurement or beverage-specific mean ethanol content estimates would improve the precision of survey alcohol assessment. PMID- 16340460 TI - Alcohol consumption among low-income pregnant Latinas. AB - BACKGROUND: Due to changing cultural norms, Latinas of childbearing age residing in the U.S. may be at increasing risk of drinking harmful levels of alcohol during pregnancy, and may also be unaware of the risks for Fetal Alcohol Spectrum Disorders associated with this behavior. We assessed the prevalence of alcohol consumption in a sample of low-income pregnant Latinas and examined risk factors for alcohol use in the periconceptional period. METHODS: As part of a larger intervention trial, a cross-sectional in-home interview study was conducted among a sample of 100 pregnant low-income Latinas receiving services from the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) in San Diego County, California. RESULTS: Fifty-seven percent of respondents indicated they were either life-time abstainers or had not consumed any alcohol in the periconceptional period. Forty-three percent reported some alcohol use in the three months prior to recognition of the current pregnancy, and 20% reported at least one binge episode of four or more standard drinks during that time frame. Five percent reported drinking seven or more drinks per week, and 8% continued drinking alcohol after recognition of pregnancy. Significant predictors of any alcohol use in the periconceptional period included English language/higher level of acculturation, younger maternal age, lower parity, higher level of education, younger age at first drink, and having ever smoked. Women who were aware of alcohol warning messages and /or had more knowledge of the Fetal Alcohol Syndrome (FAS) were significantly more likely to have consumed alcohol in the periconceptional period. Frequency of periconceptional use of alcohol did not differ between women who planned or did not plan the pregnancy. CONCLUSION: The prevalence and pattern of early pregnancy alcohol consumption in this sample of Latinas is similar to patterns noted in other race/ethnic groups in the U.S. Level of knowledge about FAS and awareness of warning messages was not protective for early pregnancy alcohol consumption, suggesting that specific knowledge was insufficient to prevent exposure or that other factors reinforce maintenance of alcohol consumption in early pregnancy. Selective interventions in low-income Latinas are warranted, and should be focused on women of reproductive age who are binge or frequent drinkers and who are at risk of becoming pregnant. PMID- 16340461 TI - Effects of chronic alcohol consumption in a visual attention task and an auditory oddball task: an event-related potential study. AB - BACKGROUND: In alcohol-dependent individuals changes in brain functioning, as measured with Event Related Potentials (ERP) have been reported. METHODS: In the present study a visual attention and an auditory oddball task were used to investigate possible differences between light, moderate, and heavy social drinkers and excessive drinkers. It was hypothesized that with increasing alcohol intake an increasing number of ERP components elicited in the visual attention task and the auditory oddball task would show diminished amplitudes. RESULTS: No differences were found between light, moderate, and heavy social drinkers. A trend for a smaller P3 amplitude in the visual attention task was found when comparing the alcohol-dependent participants with the light social drinkers. It is argued that this difference might be an effect of alcohol dependence and/or a reflection of possible unknown or undetected family history of alcohol-related disturbances. CONCLUSIONS: In the current study, even at rather large amounts of regular alcohol intake, no evidence was found for any toxic effect of social alcohol use neither in a visual attention task nor in an auditory oddball task. PMID- 16340462 TI - Ethanol administration to cystic fibrosis knockout mice results in increased fatty acid ethyl ester production. AB - BACKGROUND: Fatty acid ethyl esters (FAEE) are nonoxidative ethanol metabolites shown to produce toxic effects in the liver and pancreas in vivo and in vitro. Because alcohol-induced chronic pancreatitis is associated with mutations in the gene responsible for cystic fibrosis (CFTR), we hypothesized that CFTR dysfunction leads to increased levels of these toxic nonoxidative ethanol metabolites following alcohol administration. METHODS: Cystic fibrosis (CF) and wild-type (WT) mice were injected intraperitoneally with 1, 2, or 3 g/kg of 50% ethanol. Mice were sacrificed and the liver and pancreas removed for FAEE analysis. RESULTS: The mean FAEE concentration (pmol/g) detected in the liver of cftr mice following injection with 2 g/kg of ethanol was significantly greater than the amount detected in WT (p < 0.005). A similar trend in FAEE concentration was seen in the pancreas, but the difference was not statistically different. In both the liver and pancreas, analysis of individual FAEE species demonstrated a selective increase in ethyl oleate. CONCLUSION: These data show an association between CFTR dysfunction and qualitative and quantitative changes in FAEE in liver and pancreas upon ethanol exposure. PMID- 16340463 TI - Effects of chronic exposure to ethanol on glutamate transporter EAAT3 expressed in Xenopus oocytes: evidence for protein kinase C involvement. AB - BACKGROUND: Glutamate transporters (excitatory amino acid transporters, EAAT) regulate extracellular concentrations of glutamate, a major excitatory neurotransmitter. We reported that acute ethanol exposure increases the activity of a major neuronal EAAT, EAAT3. This effect is consistent with the general inhibitory effect of acute alcohol toxicity in the central nervous system (CNS). However, chronic ethanol exposure has CNS presentations different from acute alcohol toxicity. We hypothesize that chronic ethanol exposure will affect the EAAT3 activity differently from acute ethanol exposure. METHODS: EAAT3 was expressed in Xenopus oocytes by injection of EAAT3 mRNA. Oocytes were incubated with diluted ethanol for 24-96 hr. Using two-electrode voltage clamp, membrane currents were recorded after the application of L-glutamate. Responses were quantified by integration of the current trace and reported as microCoulombs (microC). RESULTS: Ethanol (10-100 mM) reduced EAAT3 activity in a time-dependent and reversible manner. After a 96 hr-incubation, the activity was significantly decreased compared to the control values at any concentrations tested in this study. Kinetic study demonstrated that a 96 hr-exposure to 50 mM ethanol significantly decreased Vmax (3.6 +/- 0.3 for control versus 2.6 +/- 0.3 microC for ethanol, n = 20, p < 0.05) but had no effect on Km (57.6 +/- 12.8 for control versus 51.8 +/- 16.3 microM for ethanol, n = 20, p > 0.05) of EAAT3 for glutamate. When ethanol (50 mM for 96 hr)-treated oocytes were incubated with phorbol-12-myrisate-13-acetate (50 or 100 nM for 10 min), ethanol-induced decrease in EAAT3 activity was abolished. Preincubation of the oocytes with 100 microM chelerythrine significantly decreased EAAT3 activity (1.00 +/- 0.08 for control versus 0.51 +/- 0.09 microC for chelerythrine group, n = 18-20, p < 0.05). However, there was no statistical difference among the chelerythrine, ethanol, or chelerythrine plus ethanol groups. Likewise, staurosporine (2 microM for 1 hr) significantly decreased EAAT3 activity and there was no statistical difference among the staurosporine, ethanol, or staurosporine plus ethanol groups. CONCLUSIONS: Our results show that chronic ethanol exposure decreases EAAT3 activity at clinically relevant concentrations and that this effect may be protein kinase C-dependent. Such an effect could be a neuroadaptive mechanism to overcome the inhibitory effect of ethanol on the excitatory neurotransmission. PMID- 16340464 TI - Moderate fetal alcohol exposure impairs the neurogenic response to an enriched environment in adult mice. AB - BACKGROUND: Prenatal ethanol exposure results in a spectrum of cognitive and behavioral deficits and affects an estimated thirteen percent of children born in the United States. The basis of prenatal ethanol-induced impairment of brain function has been widely studied in animal models, where significant changes in the physiological and structural plasticity of hippocampal function have been documented. Here, we explored the possibility that exposure to moderate levels of alcohol in utero might also result in long-lasting impairment of adult hippocampal neurogenesis, a novel form of plasticity that occurs throughout adulthood. METHODS: Female mice were trained to voluntarily consume 10% EtOH throughout pregnancy using the two-bottle choice paradigm, which results in moderate blood alcohol levels of approximately 121 mg/dl, as previously described (Allan et al., 2003). Offspring were exposed to standard or enriched living conditions for 8-12 weeks post-weaning. BrdU was administered at 50 mg/kg for 12 consecutive days. Mice in each housing condition were sacrificed at either 24 hrs or four weeks following the final BrdU injection, and BrdU cells within the dentate gyrus were evaluated using immuno-histochemical methods. RESULTS: Neither fetal alcohol exposure (FAE) nor enriched environment affected the number of proliferating progenitors within the subgranular zone (SGZ) of the dentate gyrus. However, FAE severely impaired the neurogenic response to enriched environment. Control mice housed in enriched environment displayed a two-fold increase in hippocampal neurogenesis, whereas FAE mice responded to enriched environment with neither enhanced progenitor survival nor enhanced neuronal differentiation. CONCLUSIONS: These observations indicate that moderate FAE results in a long term, persistent defect in neurogenic responses to behavioral challenge. PMID- 16340465 TI - Ethanol exposure and withdrawal sensitizes the rat hippocampal CA1 pyramidal cell region to beta-amyloid (25-35)-induced cytotoxicity: NMDA receptor involvement. AB - BACKGROUND: Millions of Americans suffer from Alzheimer's Disease (AD), which is characterized by significant neurological impairment and an accumulation in brain tissue of senile plaques consisting of beta amyloid (Abeta) peptide. The hippocampus, a region primarily responsible for learning and memory, appears to be particularly susceptible to AD-related injury and chronic alcohol abuse. Although certain risk factors for AD are known, it is unclear if alcohol abuse or dependence may contribute to neuropathology in AD. Recent research suggests that low-to-moderate consumption of alcohol may protect against development of AD, while alcohol dependence may increase risk of developing AD. Therefore, the current studies aimed to investigate the effects of exposure to 50 or 100 mM ethanol (EtOH) and withdrawal on hippocampal injury induced by Abeta peptide treatment. METHODS: The present studies exposed organotypic hippocampal slice cultures to 50 or 100 mM ethanol (EtOH) for 10 days, after which the slices underwent ethanol withdrawal (EWD) in the presence of varying concentrations of Abeta 25-35 (0.1, 1, 10 microM), or 35-25 (200 microM), a negative control reverse sequence peptide. Cellular injury, as evidenced by uptake of propidium iodide (PI), was assessed for each subregion of the hippocampal complex (CA1, CA3, and dentate gyrus). RESULTS: Cellular injury in the CA1 pyramidal cell layer was significantly increased during withdrawal from exposure to 100 mM, but not 50 mM, EtOH. Exposure to Abeta in ethanol-naive cultures did not produce significant cytotoxicity. However, exposure to Abeta during EWD from 100 mM produced marked increases in CA1 pyramidal cell region cytotoxicity, effects reversed by cotreatment with a nontoxic concentration of the NMDA receptor channel blocker MK 801 (20 microM). CONCLUSIONS: These data suggest that withdrawal from exposure to a high concentration of EtOH produces marked cellular injury in the hippocampus, particularly the CA1 subregion. Further, this EtOH exposure and withdrawal regimen sensitizes the hippocampus to the toxic effects of Abeta treatment in a manner reflecting over activity of NMDA receptor function. PMID- 16340466 TI - Recovery of CD4+ T Cells in HIV patients with a stable virologic response to antiretroviral therapy is associated with polymorphisms of interleukin-6 and central major histocompatibility complex genes. AB - We investigated whether polymorphisms in genes associated with HIV disease progression and/or immune activation affect CD4+ T-cell recovery in HIV patients who began combination antiretroviral therapy (ART) with advanced immunodeficiency and achieved stable control of plasma viremia. Patients with CD4 T-cell counts <300 cells/microL (n = 33) and >400 cells/microL (n = 37) on ART were compared. A multiple case-control logistic regression associated carriage of BAT1(1,2) or interleukin (IL)6-174(2,2) with low CD4 T-cell counts (P = 0.012). BAT1*2 uniquely marks the central major histocompatibility complex region of a conserved haplotype (HLA-A1,B8,BAT1*2,TNFA-308*2,DR3,DQ2). There was no association between alleles carried at CCR5Delta32, CCR5 59029, CCR5 59353, CCR2+190 (V64I), SDF1 3'UTR, IL1A+4845, IL1B+3953, IL4-589, IL10-592, IL10-R1+536, IL10-R1+1112, IL12B 3'UTR, TNFA-308, or TNFA-1031 and CD4 T-cell counts. We suggest that immune activation and/or CD4 T-cell apoptosis in HIV patients on effective ART is influenced by genetic factors. PMID- 16340467 TI - CC chemokines induce neutrophils to chemotaxis, degranulation, and alpha-defensin release. AB - We have previously shown that a Taiwanese cohort of HIV-uninfected individuals was associated with the significantly elevated levels of serum beta-chemokines, macrophage inflammatory protein (MIP-1)-alpha and MIP-beta, and RANTES. In the present study, we report that the members of this cohort have significantly greater numbers of lower buoyant-density neutrophils in their blood, which leads to further investigation of the effects of beta-chemokines on neutrophils. By electron and confocal microscopic techniques and FACScan, the results demonstrated that MIP-1alpha, MIP-beta, and/or RANTES readily activated the cells to release a large quantity of alpha-defensins in vitro through the degranulation process, which was the cause of low-buoyant-density neutrophil production. The purified neutrophils underwent chemotaxis and increased phagocytic capability when beta-chemokines were present. Only when using all 3 neutralizing antibodies for CCR1, CCR3, and CCR5 could the chemotaxis of neutrophils be inhibited completely, suggesting that these receptors are involved in transducing activating signals. Because neutrophils are the most abundant white blood cells that can be activated simultaneously to release alpha-defensins and because these proteins are antiviral, including anti-HIV, our results support the hypothesis that in addition to beta-chemokines, the innate immunity of the cohort plays a role in inhibiting the transmission of HIV. PMID- 16340468 TI - Polymorphism of RANTES chemokine gene promoter is not associated with long-term nonprogressive HIV-1 infection of more than 16 years. AB - To examine whether polymorphisms of the RANTES chemokine gene promoter are associated with long-term nonprogressive HIV-1 infection in white Spanish subjects, we performed a cross-sectional genetic association case-control study. Two-hundred sixty-seven white Spaniards were studied: 58 were HIV-1-infected long term nonprogressors (LTNPs) of more than 16 years, 109 were HIV-1-infected usual progressors (UPs), and 100 were control subjects. Three RANTES single nucleotide polymorphisms (SNPs) at positions -28C>G, -109T>C, and -403G>A were assessed. The prevalence of the CCR5Delta 32 allele was also examined. Genotyping was performed using polymerase chain reaction and automatic sequencing analysis methods. Genotype and allele frequencies between the 3 groups were compared by the chi2 test and the Fisher exact test. The distribution of allelic variants of RANTES in controls, UPs, and LTNPs, respectively, was 3%, 2%, and 5% for -28G; 4%, 2%, and 2% for -109C; and 18%, 18%, and 18% for -403A (P = not significant). The differences were still nonsignificant when we exclusively analyzed individuals not carrying the CCR5Delta32 allele. We conclude that LTNP of more than 16 years is not associated with SNPs in the RANTES gene promoter in white Spanish HIV-1 infected subjects. PMID- 16340469 TI - Is moderate HIV viremia associated with a higher risk of clinical progression in HIV-infected people treated with highly active antiretroviral therapy: evidence from the Italian cohort of antiretroviral-naive patients study. AB - OBJECTIVE: To assess the risk of clinical progression (CP) according to the duration of time spent without complete viral load (VL) suppression compared with that associated with periods of stably suppressed viremia in HIV-infected people who started highly active antiretroviral therapy (HAART) when previously naive to antiretrovirals. DESIGN: A cohort study of patients having started HAART after enrollment in the Italian Cohort of Antiretroviral-Naive Patients (ICoNA) and being followed for at least 6 months. METHODS: Person-years spent in different categories according to the VL level and the change in VL from the most recent value before the initiation of HAART were calculated. A multivariable Poisson regression model, including potential confounders, was constructed. RESULTS: A total of 3023 patients were studied. The overall rate of CP was 13.4 per 1000 person-years. Evidence for a higher risk of CP was observed for people with a current VL >10,000 copies/mL. For each year longer spent on HAART with a VL >100,000 copies/mL, a 5-fold increased risk was observed (relative risk [RR] = 5.34, 95% confidence interval [CI]: 2.83 to 1.08; P = 0.0001). An increased risk of CP in patients with current suppression <1.5 log10 copies/mL (RR = 2.34, 95% CI: 1.16 to 4.74; P = 0.02) and in those with no suppression or a VL higher than their set point (RR = 2.39, 95% CI: 1.17 to 4.89; P = 0.02) was observed compared with those with suppression of >3 log10 copies/mL, although it was not significant. Longer duration on HAART with a VL suppressed below set point seemed to confer protection against CP. CONCLUSIONS: Virologic failure to antiretroviral drugs is common. The risk of CP may remain low despite a low but detectable level of HIV viremia. PMID- 16340470 TI - Clinical presentation and course of acute hepatitis C infection in HIV-infected patients. AB - Hepatitis C virus (HCV) has become a significant source of morbidity and mortality in HIV-infected patients. However, little is known about the clinical presentation and course of acute HCV infection in this population. This study reports the outcomes of acute HCV infection in 9 HIV-infected men. Sex with men was the only reported risk factor for HCV infection in 6 of the subjects. Clinical presentation of acute HCV ranged from incidentally discovered elevated transaminases to severe liver dysfunction requiring hospitalization. At the time of HCV diagnosis, 8 of 9 patients had CD4+ counts >250 cells/mm(3), and 6 had HIV viral loads of < or =5000 copies/mL. Eight patients were receiving antiretroviral therapy. Outcome of these acute HCV infections varied. Five patients experienced virologic clearance, 2 in whom virus cleared spontaneously and 3 who were treated with pegylated interferon and ribavirin. Four patients developed chronic infection, one of whom had a relapse during HCV treatment and 3 of whom were untreated. All 4 patients to whom HCV therapy was administered experienced significant anemia or neutropenia, necessitating dose reduction or support with growth factors. Prompt recognition of acute HCV infection may minimize antiretroviral treatment interruption and will allow early treatment, which may improve virologic clearance. Unexplained transaminase elevations in HIV-infected patients, including men who have sex with men, should trigger an evaluation for acute HCV infection. PMID- 16340471 TI - Assessment of drug-drug interactions between tenofovir disoproxil fumarate and the nonnucleoside reverse transcriptase inhibitors nevirapine and efavirenz in HIV-infected patients. AB - BACKGROUND: Tenofovir disoproxil fumarate (DF) has been studied in combination with efavirenz in healthy volunteers and no interaction was found. No data are available on the possible interaction of tenofovir DF with nevirapine and efavirenz in HIV-infected patients. In this study the combination of nevirapine 200 mg twice daily with tenofovir DF 300 mg once daily and nevirapine 400 mg once daily with tenofovir DF 300 mg once daily were compared with nevirapine twice daily or once daily without tenofovir DF in HIV-infected patients. Furthermore, the combination of efavirenz 600 mg and tenofovir DF 300 mg once daily was compared with use of efavirenz 600 mg once daily only. METHODS: Data were retrospectively collected from routine therapeutic drug monitoring plasma samples. Nevirapine, efavirenz, and tenofovir plasma levels and tenofovir concentration ratios were analyzed. The concentration ratio represents the measured plasma concentration compared with the time-adjusted average concentration, as measured in a reference population. Six different groups were studied: 200 mg nevirapine twice daily, 400 mg nevirapine once daily, 600 mg efavirenz once daily, all without tenofovir DF (groups 1, 2, and 3, respectively), and the same groups with the drugs combined with tenofovir 300 mg once daily (groups 4, 5, and 6, respectively). RESULTS: Plasma samples were evaluable for 272, 18, 126, 32, 94, and 118 patients in the groups 1-6, respectively. No differences were found in plasma levels for tenofovir, nevirapine, and efavirenz for either of the combinations studied. Addition of tenofovir DF to efavirenz or nevirapine in HIV-infected patients does not influence the plasma levels of nevirapine or efavirenz. Furthermore, nevirapine and efavirenz have no effect on tenofovir plasma levels or tenofovir concentration ratios. CONCLUSION: Efavirenz or nevirapine can be coadministered with tenofovir DF in HIV-infected patients without dose modifications. PMID- 16340472 TI - Clinician-delivered intervention during routine clinical care reduces unprotected sexual behavior among HIV-infected patients. AB - OBJECTIVE: To evaluate the effectiveness of a clinician-delivered intervention, implemented during routine clinical care, in reducing unprotected sexual behavior of HIV-infected patients. DESIGN: A prospective clinical trial comparing the impact of a clinician-delivered intervention arm vs. a standard-of-care control arm on unprotected sexual behavior of HIV-infected patients. SETTING: The 2 largest HIV clinics in Connecticut. PARTICIPANTS: A total of 497 HIV-infected patients, aged > or =18 years, receiving HIV clinical care. INTERVENTION: HIV clinical care providers conducted brief client-centered interventions at each clinical encounter that were designed to help HIV-infected patients reduce unprotected sexual behavior. MAIN OUTCOME MEASURES: Unprotected insertive and receptive vaginal and anal intercourse and unprotected insertive oral sex; unprotected insertive and receptive vaginal and anal intercourse only. RESULTS: HIV-infected patients who received the clinician-delivered intervention showed significantly reduced unprotected insertive and receptive vaginal and anal intercourse and insertive oral sex over a follow-up interval of 18 months (P < 0.05). These behaviors increased across the study interval for patients in the standard-of-care control arm (P < 0.01). For the measure of unprotected insertive and receptive vaginal and anal sex only, there was a trend toward a reduction in unprotected sex among intervention arm participants over time (P < 0.09), and a significant increase in unprotected sex in the standard-of-care control arm (P < 0.01). CONCLUSIONS: A clinician-delivered HIV prevention intervention targeting HIV-infected patients resulted in reductions in unprotected sex. Interventions of this kind should be integrated into routine HIV clinical care. PMID- 16340473 TI - Reasons for modification of generic highly active antiretroviral therapeutic regimens among patients in southern India. AB - OBJECTIVE: To describe reasons for modification and discontinuation of antiretroviral regimens in association with adverse events (AEs), treatment failure, and cost among patients in southern India. METHODS: Secular trends of patients initiating highly active antiretroviral therapy (HAART) between January 1996 and October 2004 at a tertiary HIV referral center in India were analyzed using a previously validated natural history database. RESULTS: All previously antiretroviral therapy-naive patients who initiated HAART (N = 1443) and had at least 1 follow-up visit were evaluated. The median CD4 count at the time of initiating HAART was 108 cells/microL. The most common first-line regimens were stavudine (d4T) plus lamivudine (3TC) plus nevirapine (NVP) (63%), zidovudine (AZT) plus 3TC plus NVP (19%), d4T plus 3TC plus efavirenz (EFV) (9%), and AZT plus 3TC plus EFV (4%). Twenty percent of patients modified their first-line regimen. The most common reason for modifying therapy was the development of an AE (64%), followed by cost (19%) and treatment failure (14%), with median times to modify therapy being 40, 151, and 406 days, respectively. Common AEs were itching and/or skin rash (66%), hepatotoxicity (27%), and anemia (23%). Nine percent of patients discontinued therapy entirely after a median duration of 124 days, primarily because of cost (64%). CONCLUSION: The most common reason for modifying therapy was the occurrence of AEs, whereas cost was the most common reason for discontinuing therapy. Despite increasing access to lower cost generic HAART in India, even less expensive and more tolerable first-line regimens and cost-effective treatment monitoring tools need to be introduced to achieve better treatment outcomes and access in resource-constrained settings. PMID- 16340474 TI - Utilization and access to antiretroviral genotypic resistance testing and results within the US Department of Veterans Affairs. AB - OBJECTIVE: We sought to characterize variation in the use of HIV genotypic resistance tests and how results were reported. METHODS: Clinicians and laboratory managers at all Veterans Affairs (VA) medical centers were asked to complete a survey in March 2003 regarding HIV resistance testing practices. RESULTS: Surveys from 131 of 150 sites were returned. Forty-eight percent of HIV clinicians indicated that US Department of Health and Human Services guidelines were the usual basis for ordering tests. Although between 12% and 31% of respondents indicated that they always, sometimes, seldom, or never ordered resistance tests in patients with acute or chronic HIV infection, >70% ordered tests in adherent patients with treatment failure. Among the 32 centers with >200 patients in care, 13 +/- 8 (mean +/- standard deviation) tests were performed per 100 patients in care during 2002. Forty-nine percent of clinicians said that tests were helpful, but only 33% expressed confidence in using test results. Only 40% of sites entered results in the VA electronic medical record. CONCLUSION: Ordering patterns for HIV resistance tests differed significantly among VA sites. A minority of clinicians indicated confidence in the use of test results. A consistent system to capture and present complete results was absent. PMID- 16340475 TI - Liver fibrosis in HIV-positive patients with hepatitis C virus: role of persistently normal alanine aminotransferase levels. AB - BACKGROUND: Liver fibrosis requiring treatment in HIV/hepatitis C virus (HCV) coinfected patients with persistently normal alanine aminotransferase (ALT) values (PNAL) is currently not well defined; in this study clinical and histologic features of PNAL were compared with those of subjects with elevated ALT (EAL). METHODS: A total of 326 liver biopsies of HIV/HCV-coinfected patients, performed from 1997-2003, were retrospectively identified. Subjects with at least 3 consecutive normal ALT determinations during a prebiopsy follow-up of 12 months were grouped as PNAL (24 patients) and compared with EAL subjects (302 patients). Liver biopsy was classified with the modified Ishak score. RESULTS: Age, HCV viral load, and genotype, CD4 T-cell count, and antiretroviral drugs did not show a statistical difference between the 2 groups. Statistical significance was found when comparing mean grading (1.4 +/- 1.8 vs. 7.2 +/- 2.6, P < 0.0001) and staging (1.4 +/- 1.79 vs. 2.5 +/- 1.7, P < 0.0003) between PNAL and EAL subjects. The proportion of PNAL patients fulfilling histologic criteria for anti-HCV treatment (25% with stage 2-6) was also significantly different from EAL subjects (69%; P = 0.0001). At multivariate analysis, only age, CD4 count (>500 vs. < or =500 cells/mL), and patient's group (EAL vs. PNAL) were found to be independently associated with a fibrosis score of > or =2. CONCLUSION: Liver fibrosis requiring treatment was found in 25% of HIV/HCV-coinfected subjects with PNAL values. PMID- 16340476 TI - Genotypic resistance and immunologic outcomes among HIV-1-infected women with viral failure. AB - OBJECTIVES: To describe the prevalence of specific protease inhibitor (PI) and nonnucleoside reverse transcriptase inhibitor (NNRTI) resistance mutations and the relationship between the presence of these mutations and immunologic outcomes following PI/NNRTI initiation among a cohort of HIV-1-infected women. METHODS: Viral genotypic resistance testing was done for 366 women enrolled in the Women's Interagency HIV Study at the visit immediately prior to 1st reported use of PI or NNRTI (baseline) and at the visit approximately 1 year after PI/NNRTI initiation. We modeled the changes in CD4+ T-cell counts and HIV RNA levels approximately 1 year after therapy initiation as a function of baseline and follow-up markers, type of antiretroviral therapy used, and resistance mutations. RESULTS: At baseline, 52% of women showed only nucleoside reverse transcriptase inhibitor (NRTI) mutations, 38% showed no mutations, and 10% showed PI or NNRTI mutations. Only 40% of women showed viral response (HIV-1 RNA < or = 80 copies/mL) 1 year after initiating a PI or NNRTI. Among those without a viral response, 54% developed PI or NNRTI mutations. NNRTI (among those with baseline NRTI mutations) and PI resistance mutations were associated with better CD4+ cell count changes (mean increase of 118 cells/mm3 and 64 cells/mm3, respectively, as compared with viral nonresponders with no PI or NNRTI mutations). CONCLUSIONS: In this population-based cohort, virologic failure with PI or NNRTI resistance was common. Viremia with these resistance mutations was associated with preserved CD4+ T-cell count responses, providing evidence of reduced virulence or viral fitness. PMID- 16340477 TI - High HIV prevalence among a high-risk subgroup of women attending sexually transmitted infection clinics in Pune, India. AB - OBJECTIVE: To investigate changes over a decade in prevalence and correlates of HIV among high-risk women attending sexually transmitted infection (STI) clinics in Pune, India, who deny a history of commercial sex work (CSW). DESIGN: Cross sectional. METHODS: From 1993 to 2002, 2376 women attending 3 STI clinics in Pune were offered HIV screening. Women who denied CSW were included (n = 1020). RESULTS: Of 1020 women, 21% were HIV infected. The annual HIV prevalence increased from 14% in 1993 to 29% in 2001-2002 (P < 0.001). The change in HIV prevalence over time was paralleled by changes in clinic visitor characteristics; in later periods, women were older, more often employed, less likely to be currently married, and more likely to report condom use. In multivariate analysis, factors independently associated with HIV were calendar period (adjusted odds ratio [AOR], 1.9 for 1997-1999 vs. 1993-1996; 95% CI, 1.2-3.0; AOR, 2.3 for 2000-2002 vs. 1993-1996; 95% CI, 1.5-3.6), lack of formal education (AOR, 2.0; 95% CI, 1.4-2.9), having been widowed (AOR, 3.1; 95% CI, 1.6-6.1), current employment (AOR, 1.8; 95% CI, 1.2-2.6), and genital ulcer disease on examination (AOR, 1.8; 95% CI, 1.2-2.7). CONCLUSIONS: Women attending STI clinics in India who deny a history of CSW represent a small, hidden subgroup, likely put at risk for HIV because of high-risk behavior of their male partners, generally their husbands. Educational and awareness efforts that have targeted other subgroups in India (men and CSWs) should also focus on these hard-to-reach women. Risk reduction in this subgroup of Indian women would also be expected to reduce perinatal infections in India. PMID- 16340478 TI - HIV-related risk behaviors in Cambodia and effects of mobility. AB - OBJECTIVES: To study HIV risk behaviors in different population groups, linkages to bridge populations, and to examine factors affecting such behaviors and links. METHODS: Ten population groups in 4 provinces were surveyed. Stratified random cluster sampling was used, and interviews were conducted to provide information on sociodemographic characteristics, mobility, and risk behaviors. The groups surveyed were female sex workers (FSWs), household men and women, youths in vocational training, and men with high-mobility occupations (fishermen, mototaxi drivers, police, military, casino workers, and deminers). The total number surveyed was 3848. RESULTS: The proportion reporting sex in the past year with FSWs differed sharply between male groups ranging from 20% to 51% in the high mobility groups and 5% to 10% in the other groups. Noncommercial sex varied less by group. Consistent condom protection (always used condoms in the past 3 months) with FSWs was high (>85% for most groups). However, condom use was significantly less with noncommercial partners, a high proportion of whom complained about a lack of condom availability. For the different male groups, travel away from home >1 month in the past year was a strong independent determinant of both sex with FSWs and noncommercial sex. Casual sex was more common in young unmarried men. Women in the general population did not report casual sex, but 41% of them were "worried about being infected by their husbands." CONCLUSIONS: The results suggest mobility is a strong determinant of casual sex. Although FSWs may still act as an important bridge for HIV transmission in Cambodia, noncommercial sex is becoming increasingly important due to the relatively low condom use in such relationships. PMID- 16340479 TI - Perinatal HIV counseling and rapid testing in Tijuana, Baja California, Mexico: seroprevalence and correlates of HIV infection. AB - OBJECTIVE: To evaluate the acceptance of counseling and rapid HIV testing and to determine the associated risk factors for HIV infection in pregnant women in Baja California, Mexico. METHODS: Pregnant women attending Tijuana General Hospital who consented to participate in the study had blood drawn for a rapid HIV test (Determine HIV-1/2; Abbott Diagnostics, North Chicago, IL). A confirmatory enzyme immunoassay and Western blot were performed and demographic and risk factor data were obtained. RESULTS: From March to November 2003, 1529 (92.5%) of 1653 women who sought prenatal care and 1068 (95.2%) of 1122 women in labor consented to participate. HIV seroprevalence was significantly higher among women screened during labor (12/1068, 1.12%) compared with those seeking prenatal care (5/1529, 0.33%). HIV-infected women were significantly more likely to use injection drugs (12% vs. 1%, P = 0.02), "other" drugs, including methamphetamine, marijuana, and cocaine (65% vs. 6%, P < 0.001), to have more sex partners (3.6 vs. 2.6, P = 0.0002), to not have received prenatal care (41% vs. 13%, P = 0.03), and to have a spouse/partner who used injection drugs (36% vs. 4%, P < 0.001) or "other" drugs (73% vs. 23%, P < 0.001). In multivariate regression analysis, use of methamphetamine (adjusted odds ratio, 17.8, 95% CI, 5.6-56) was independently associated with the risk of HIV infection. CONCLUSIONS: These findings indicate a wide acceptance of HIV counseling and testing and document a higher HIV seroprevalence among pregnant women delivering at Tijuana General Hospital than current established estimates in Mexico. PMID- 16340481 TI - Coinfection with HIV-1 and human T-Cell lymphotropic virus type II in intravenous drug users is associated with delayed progression to AIDS. AB - Human T-cell lymphotropic virus (HTLV) type II has spread among intravenous drug users (IDUs), many of whom are coinfected with HIV-1. We have investigated the rate of HTLV-II infection in 3574 Italian IDUs screened for HIV-1, HTLV-I, and HTLV-II from 1986 to the present. HTLV-II proviral load was determined by a real time polymerase chain reaction specifically designed for tax amplification. The frequency of HTLV-II infection was 6.7% among HIV-1-positive subjects and 1.1% among HIV-1-negative subjects (P < 0.0001). For examination of AIDS progression, a group of 437 HIV-1-monoinfected subjects and another group of 96 HIV-1/HTLV-II coinfected subjects were monitored. Enrollees were matched at entry by CD4 cell counts and followed for an average of 13 years. HIV-1/HTLV-II coinfection was associated with older age (P < 0.0001) and higher CD4 (P < 0.0001) and CD8 (P < 0.001) cell counts compared with monoinfected IDUs. The number of long-term nonprogressors for AIDS was significantly higher (P < 0.0001) among coinfected patients (13 [13.5%] of 96 patients) than HIV monoinfected patients (5 [1.1%] of 437 patients), showing that HTLV-II exerts a protective role. An increased incidence of liver disease and hepatitis C virus positivity among coinfected IDUs was observed. Five coinfected subjects undergoing antiretroviral therapy showed a significant (P < 0.05) increase in HTLV-II proviral load concomitant to a decrease in HIV-1 viremia, suggesting that the treatment is ineffective against HTLV-II infection. PMID- 16340482 TI - Correlates and trend of HIV prevalence among female sex workers attending sexually transmitted disease clinics in Pune, India (1993-2002). AB - In India, substantial efforts have been made to increase awareness about HIV/AIDS among female sex workers (FSWs). We assessed the impact of awareness regarding safe sex in a cohort of FSWs by studying trends in HIV prevalence, sexually transmitted diseases (STDs), and risk behaviors measured from 1993 to 2002 in Pune, India. A total of 1359 FSWs attending 3 STD clinics were screened for HIV infection, and data on demographics, sexual behaviors, and past and current STDs were obtained. The overall HIV prevalence among FSWs was 54%. Not being married (adjusted odds ratio [AOR] = 1.74, 95% confidence interval [CI]: 1.17 to 2.59), being widowed (AOR = 2.10, 95% CI: 1.16 to 3.80), inconsistent condom use (AOR = 1.60, 95% CI: 1.02 to 2.50), clinical presence of genital ulcer disease (GUD; AOR = 1.66, 95% CI: 1.07 to 2.56), and genital warts (AOR = 4.70, 95% CI: 1.57 to 14.08) were independently associated with HIV infection among FSWs. The prevalence of HIV remained stable over 10 years (46% in 1993 and 50% in 2002; P = 0.80). The prevalence of GUD decreased over time (P < 0.001), whereas that of observed genital discharge remained stable. Reported consistent condom use as well as the proportion of FSWs who refused sexual contact without condoms increased over time (P < 0.001). These data collectively suggest that safe sex interventions have had a positive impact on FSWs in Pune, India. PMID- 16340483 TI - Predictors of nonadherence to single-dose nevirapine therapy for the prevention of mother-to-child HIV transmission. AB - BACKGROUND: Adequate adherence is required for prevention of mother-to-child transmission of HIV (pMTCT) programs to be effective. We investigated predictors and extent of nonadherence to single-dose nevirapine. METHODS: Data on nevirapine intake and possible predictors were collected among 760 HIV-positive women with liveborn babies enrolled in a study in Lusaka, Zambia. RESULTS: Most (94%) women took nevirapine before delivery, and most (91%) newborns received it soon after delivery. Maternal nonadherence was associated with home births (odds ratio [OR]: 3.2; 95% confidence interval [CI]: 1.3 to 7.4), no high school education (OR: 2.4; 95% CI: 1.1 to 5.3), and low newborn birth weight (OR: 4.6; 95% CI: 1.3 to 20.1). Disclosure of HIV status and couples counseling was only associated with adherence among home births. Failure to administer nevirapine to the newborn was associated with birth at the tertiary hospital (OR: 7.2; 95% CI: 3.7 to 13.8), lower 5-minute Apgar scores (OR: 0.5; 95% CI: 0.4 to 0.7), and neonatal death (OR: 5.8; 95% CI: 2.0 to 16.3). CONCLUSIONS: Excellent adherence to single-dose nevirapine for pMTCT can be achieved. Nonadherence seems to be affected by place of birth and by poor health status of the newborn. Procedures to ensure that viable yet ill neonates receive nevirapine should be part of clinical protocols and training within pMTCT programs. PMID- 16340484 TI - The impact of sex partners' HIV status on HIV seroconversion in a prospective cohort of injection drug users. AB - OBJECTIVE AND DESIGN: The identification of individuals at highest risk of HIV infection is critical for targeting prevention strategies. We evaluated the HIV status of the sex partners of injection drug users (IDUs) and rates of subsequent HIV seroconversion among a prospective cohort study of IDUs. METHODS: We performed an analysis of the time to HIV infection among baseline HIV-negative IDUs enrolled in the Vancouver Injection Drug Users Study. IDUs were stratified based on whether or not they reported having an HIV-positive sex partner. Kaplan Meier methods were used to estimate cumulative HIV incidence rates, and Cox regression was used to determine adjusted relative hazards (RHs) for HIV seroconversion. RESULTS: Of 1013 initially HIV-negative IDUs, 4.8% had an HIV positive partner at baseline. After 18 months, the cumulative HIV incidence rate was significantly elevated among those who reported having an HIV-positive sex partner (23.4% vs. 8.1%; log-rank P < 0.001). In a Cox regression model adjusting for all variables that were associated with the time to HIV infection in univariate analyses, including drug use characteristics, having an HIV-positive sex partner (RH = 2.42 [95% confidence interval: 1.30 to 4.60]; P = 0.005) remained independently associated with time to HIV seroconversion. CONCLUSIONS: Having an HIV-positive sex partner was strongly and independently associated with seroconversion after adjustment for risk factors related to drug use. Our findings may aid public health workers in their efforts to identify IDUs who should be targeted with education and prevention efforts and indicate the need for ongoing development of prevention interventions for IDU sex partners who are HIV discordant. PMID- 16340485 TI - Interaction between atazanavir and fosamprenavir in the treatment of HIV-infected patients. PMID- 16340486 TI - A series of skin and soft tissue infections due to methicillin-resistant Staphylococcus aureus in HIV-infected patients. PMID- 16340487 TI - Patients experiencing early virologic failure on a protease inhibitor- or nonnucleoside reverse transcriptase inhibitor-based initial regimen containing a thymidine analogue and Lamivudine can be successfully treated with a quadruple nucleoside regimen. PMID- 16340488 TI - Association between malaria and CD4 cell count decline among persons with HIV. PMID- 16340489 TI - Carotid endarterectomy for transient monocular visual loss and other ocular ischemic conditions. PMID- 16340480 TI - Transmission of cell-free and cell-associated HIV-1 through breast-feeding. AB - BACKGROUND: Transmission through breast-feeding is an important cause of infant HIV-1 infections in developing countries; however, its mechanism remains largely unknown. We have explored the association between cell-free virus (CFV) and cell associated virus (CAV) levels in breast milk (BM), as reflected by viral RNA and proviral DNA, respectively, and the risk of infant HIV-1 infection after 6 weeks postpartum. METHODS: Sixty-one HIV-positive mothers who transmitted HIV-1 by BM were matched to 61 HIV-positive nontransmitting mothers based on their infant's age at sample collection. CFV and CAV were quantified in a single milk specimen per mother preceding the infant's first HIV-positive result. RESULTS: After adjusting for maternal CD4 cell counts and disease stage, each 10-fold increase in CFV or CAV load was associated with an almost 3-fold increase in BM transmission. Whereas CAV load was predictive of transmission before and after 9 months postpartum, CFV was a significant predictor of transmission occurring only after 9 months. Phylogenetic analyses of the C2 to C5 env region showed that 85% of infants (11 of 13 infants) harboring viruses that clustered with CFV in their mother's milk were infected after 9 months postpartum. CONCLUSION: A reduction in milk CAV and CFV loads might significantly decrease HIV-1 transmission by breast feeding. PMID- 16340490 TI - Ophthalmic manifestations in 18 patients with botulism diagnosed in Porto, Portugal between 1998 and 2003. AB - BACKGROUND: Botulism is a rare but potentially lethal disease in which ophthalmic signs and symptoms are among the very earliest manifestations. The aim of this study was to investigate the epidemiological and clinical features of botulism infected patients admitted to a general hospital in Porto, Portugal. METHODS: We performed a retrospective chart review of all botulism patients admitted to Sao Joao Hospital between January 1998 and January 2003. We excerpted data on epidemiology, ophthalmic and non-ophthalmic manifestations, and treatment. RESULTS: We identified 18 patients in nine registered outbreaks. In two patients (11%), ophthalmic manifestations preceded systemic manifestations; in six patients (33%), ophthalmic and systemic manifestations occurred simultaneously; in ten patients (56%), systemic manifestations occurred first. Ophthalmologists had examined only seven patients and made the correct diagnosis in five. The most common ocular symptoms were blurred near vision (100%), blurred distant vision (94%), and diplopia (44%). Accommodation impairment was documented in all seven patients examined by ophthalmologists. CONCLUSIONS: Ophthalmic manifestations were among the earliest and most prominent manifestations of botulism in this series, as in earlier reports. The diagnosis should be suspected when impaired accommodation and gastrointestinal symptoms occur together. PMID- 16340491 TI - Ocular ischemic syndrome after occlusion of both external carotid arteries. AB - The ocular ischemic syndrome (OIS) has been reported in association with high grade stenosis or occlusion of the common carotid artery (CCA) or internal carotid artery (ICA) but never with high-grade stenosis or occlusion of the external carotid artery (ECA) alone. We describe two patients who developed OIS with bilateral occlusion of the ECAs yet patent CCAs and ICAs. In one case, unilateral OIS followed consecutive bilateral carotid endarterectomies. In the other case, OIS developed spontaneously OU but was exacerbated in one eye after ipsilateral carotid endarterectomy (CE) in the setting of pre-existing contralateral ECA occlusion. In some individuals, the ECA is the primary source of arterial blood flow to the eye. Because of this fact, the endarterectomy surgeon must avoid causing ECA occlusion by meticulously removing not only the ICA plaque, but also the entire ECA plaque. PMID- 16340492 TI - Recognition of objects in non-canonical views: a functional MRI study. AB - BACKGROUND: The neural correlate of object recognition in non-canonical views is uncertain, but there is evidence for involvement of neural pathways, possibly separate from those used for object recognition in canonical views. METHODS: Boxcar functional MRI (fMRI) techniques were used to detect neural activity while eight normal subjects were instructed to identify digital photographs of objects in non-canonical and canonical orientations. RESULTS: The right angular gyrus, the left inferior temporal gyrus, and the right cerebellum showed significant fMRI activity during non-canonical as opposed to canonical viewing. CONCLUSIONS: Subjects recognizing objects in non-canonical orientations engage in a process separate from, or in addition to, the process used in recognizing objects in canonical orientations. PMID- 16340493 TI - Benign essential blepharospasm: risk factors with reference to hemifacial spasm. AB - BACKGROUND: To identify risk factors associated with benign essential blepharospasm (BEB) with reference to hemifacial spasm (HFS). Persons with BEB and HFS experience similar physical symptoms, yet the two disorders have different etiologies. METHODS: Patients with BEB (n = 159) or HFS (n = 91) were identified from two large neuro-ophthalmology clinics. Demographic, medical, behavioral, and psychological characteristics were obtained from chart review and a telephonic survey questionnaire. RESULTS: The average age of BEB and HFS was 66 years. Most patients in both groups were retired, white, and female. BEB patients were more than two times as likely to meet the diagnostic criteria for generalized anxiety disorder than HFS patients (odds ratio, 2.13; 95% confidence interval, 1.22-3.72). There was no difference between the two groups regarding demographics, smoking, a family history of dystonia, Parkinson disease, Bell palsy, Tourette disorder, obsessive compulsive symptoms, history of head trauma, alcohol use, or caffeine consumption. CONCLUSIONS: As compared to HFS, BEB was significantly more often associated with generalized anxiety disorder. Given the similarity of other clinical features of these two disorders, it is reasonable to conclude that anxiety is a cause not a consequence of BEB. Contrary to previous studies, BEB was not associated with obsessive-compulsive symptoms, head trauma, Parkinson disease, Bell palsy, Tourette disorder, or lack of smoking. PMID- 16340495 TI - Primary melanoma of the sphenoid sinus presenting with a third cranial nerve palsy. AB - An 83-year-old woman presented with right trigeminal paresthesias, upper lid ptosis, and diplopia and was found to have a right third cranial nerve palsy. MRI disclosed a right sphenocavernous mass with speckled high signal on pre-contrast T1 and correspondingly low signal on T2. Endoscopic sphenoidotomy revealed malignant melanoma. The patient underwent radiation therapy with complete and protracted resolution of the palsy. An extensive evaluation for a primary source was negative. The combination of high T1 signal and low T2 signal in a mass should suggest melanin. An exceedingly rare condition, this is the first English report of its presentation with a third cranial nerve palsy. PMID- 16340494 TI - Recurrent neuroretinitis in an adolescent with ulcerative colitis. AB - Neuroretinitis refers to an optic neuropathy in which optic disc edema is accompanied by peripapillary or macular hard exudates. Most cases involve a single episode and have no associated systemic abnormalities. In rare instances, neuroretinitis may be recurrent and lead to progressive visual loss. We describe a patient with recurrent neuroretinitis who concurrently developed ulcerative colitis, the first report of this association. PMID- 16340496 TI - Sphenoid sinus mucocele presenting as a third cranial nerve palsy. AB - A 90-year-old woman developed an acute left third cranial nerve palsy. Brain imaging revealed a left sphenoid sinus mucocele. Endoscopic marsupialization of the mucocele led to complete resolution of the third cranial nerve palsy. PMID- 16340497 TI - Erectile dysfunction drugs and non-arteritic anterior ischemic optic neuropathy: is there a cause and effect relationship? AB - The recent reports of non-arteritic anterior ischemic optic neuropathy (NAION) occurring shortly after ingestion of erectile dysfunction agents have raised the question of whether these agents have a cause-and-effect relationship to NAION. The nature of optic nerve head blood flow and the various factors that influence it, the systemic vascular effects of these agents, and the clinical features of NAION lead me to believe that these agents are contributory factors. Patients with the appropriate risk factors should, therefore, be warned of this possibility and advised to refrain from using these agents. PMID- 16340498 TI - Carotid endarterectomy for ophthalmic manifestations: is it ever indicated? AB - Four ophthalmic manifestations make up a substantial proportion of the indications for carotid endarterectomy (CE). They include transient monocular visual loss (TMVL), ocular ischemic syndrome (OIS), retinal artery occlusion (RAO), and asymptomatic Hollenhorst plaque. Critical review of the literature shows that the evidence to support the efficacy of CE in these four settings is tenuous. PMID- 16340499 TI - Carotid endarterectomy for ophthalmic manifestations: what do we do? PMID- 16340500 TI - The management of transient monocular visual loss. PMID- 16340501 TI - Cerebral perfusion imaging. AB - There are multiple imaging techniques available to assess cerebral perfusion, including positron emission tomography (PET), xenon computed tomography (XeCT), single photon emission computed tomography (SPECT), perfusion-weighted MRI (PWI), and perfusion computed tomography (PCT). Current interest has focused mainly on their use in the setting of acute brain ischemia. Perfusion imaging may be able to distinguish infarcted from salvageable ischemic tissue as a guide to treatment. Perfusion techniques may also be helpful in cases of chronic ischemia, post-subarachnoid hemorrhage vasospasm, trauma, and contemplated therapeutic carotid artery occlusion. PMID- 16340502 TI - The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), Fort Lauderdale, Florida, May 1-4, 2005. PMID- 16340503 TI - The 57th Annual Meeting of the American Academy of Neurology, Miami, Florida, April 9-16, 2005. PMID- 16340504 TI - 7th Meeting of the European Neuro-Ophthalmology Society (EUNOS), Moscow, Russia, June 26-29, 2005. PMID- 16340505 TI - Brown syndrome associated with morning glory syndrome. PMID- 16340506 TI - Teaching health departments: new variations on an old theme. PMID- 16340507 TI - Increasing smoking cessation at the population level: thinking outside the box. PMID- 16340508 TI - Academic health departments: from theory to practice. AB - In August of 2003, 23 institutions submitted proposals to build closer ties between state and local public health departments and schools of public health in response to a solicitation from the Association of Schools of Public Health supported by the Centers for Disease Control and Prevention. This article describes the strategies proposed to build collaboration between public health academia and practice. Qualitative analysis discerned five principal approaches: the development of comprehensive planning processes; reform of the way practica are planned and implemented; the identification and nurturing of boundary spanning individuals in academia and health agencies; the fostering of new approaches to joint research; and workforce development programs. Major themes that emerged included the importance of achieving a balance of power between academic and health department partners; the need to address cultural differences between institutions; a conviction that efforts at institutional change require both strong leadership and the cultivation of boundary spanners farther down the chain of command; and the idea that prospects for success may be improved if faculty and practitioners have tangible incentives to collaborate. PMID- 16340509 TI - Demonstrating excellence in practice-based teaching for public health. AB - Demonstrating Excellence in Practice-based Teaching for Public Health is a report intended to provide a resource for practice-based teaching of public health and includes a brief explanation of terms and practices, as well as suggestions on methodologies for implementation. No comparable resource currently exists that assists faculty and practice partners to recognize, implement, and promote practice-based teaching. This article summarizes findings from the report, including an explanation of practice-based teaching, its guiding principles, practical approaches, and recommendations on sustaining and advancing partnerships for professional public health education and training. PMID- 16340510 TI - Partnership for preparedness: a model of academic public health. AB - The New York City Department of Health and Mental Hygiene and the Columbia University Mailman School of Public Health's National Center for Disaster Preparedness undertook a collaborative project to establish a model academic health department. The goals were to increase student participation at the health department, increase faculty participation in health department activities, and facilitate health department faculty appointments at the school. As a result, 17 students were placed in full-time summer research projects designed by health department staff specifically for the project, 154 health department staff attended a series of six lectures presented by faculty, and five health department professionals applied for academic appointments at the school. The benefits of the efforts toward establishing an academic health department extend to all areas of public health practice, including those of preparedness. PMID- 16340511 TI - More practice, but still not perfect. PMID- 16340512 TI - Health departments and medical schools. PMID- 16340513 TI - Local health department and academic partnerships: education beyond the ivy walls. AB - The National Association of County and City Health Officials (NACCHO) is the national organization representing local health departments. NACCHO supports efforts that protect and improve the health of all people and all communities by promoting national policy, developing resources and programs, seeking health equity, and supporting effective local public health practice and systems. PMID- 16340514 TI - Reach, efficacy, and cost-effectiveness of free nicotine medication giveaway programs. AB - In an effort to increase access to evidence-based smoking cessation therapies, regional tobacco control programs in New York State implemented different interventions to make free nicotine patches and gum available to smokers wishing to quit. In one region, eligible smokers were sent a voucher redeemable at a local pharmacy for a 2-week supply of either nicotine patches or gum. In other regions, smokers received either a 1-week supply or a 2-week supply of nicotine patches sent to their home. In New York City, eligible smokers received a 6-week supply of nicotine patches and a follow-up phone call. All of the programs utilized the state's Smokers' Quitline to screen and register eligible smokers for the free medication. The reach of the different programs was evaluated by computing the proportion of eligible smokers within a given area enrolled in the program and tracking call volume to the Quitline before, during, and after the free giveaway promotions. Efficacy was evaluated by a telephone follow-up survey of program participants conducted 4 months after enrollment to measure use of the medications and smoking behavior. The quit rate of program participants was contrasted with the quit rate computed from an earlier follow-up survey of Quitline callers who were not provided nicotine replacement therapy (NRT). Free nicotine patches or gum was sent to 40,090 smokers representing about 2.9 percent of eligible heavy smokers (10+ cigarettes per day) in the state. In each time period and location where free NRT was offered, call volume to the Quitline increased dramatically. Quit rates varied in relationship to the supply of NRT sent to participants, but in all cases was higher than the quit rate observed among smokers not sent NRT (21%-35% vs 12%). The offer of free NRT appears to be a cost-effective method to induce large numbers of smokers to make a quit attempt. PMID- 16340515 TI - Financial incentives to promote smoking cessation: evidence from 11 quit and win contests. AB - In an effort footline to motivate smokers across New York State to stop smoking, tobacco control programs across the state held 'Quit and Win' incentive-based stop-smoking contests. These contests encouraged smokers to make a quit attempt by offering a chance to win cash prize (usually 1,000 dollars) for successfully stopping smoking for at least 1 month. Between 2001 and 2004, 11 different Quit and Win Contests involving 5,504 adult smokers were sponsored in different communities across New York State. Follow-up surveys were conducted 4 to 6 months after each contest ended to evaluate participants' success in quitting smoking. Expenditures for promoting contests varied from a high of 91,441 dollars to a low of 4,345 dollars, with a median of 25,928 dollars. An average of 0.55 percent of smokers was recruited to join contests across the 11 communities. Among smokers who enrolled in a contest, 9 out of 10 reported making a quit attempt, and between 53 percent and 72 percent reported quitting for the full month of the contest. At 4 to 6 months follow-up, self-reported quit rates (7-day point prevalence) among contestants ranged from 22 percent to 49 percent, with an average of 31 percent. Based on a statewide population survey, 8 of the 11 programs showed quit rates that were significantly higher (P < .001 by Wilcoxon rank-sum test) than the estimated quit rate of 21 percent seen among smokers making a quit attempt in the past year. This study shows that for a relatively modest investment of resources, thousands of smokers can be recruited to make a serious quit attempt, with many remaining smoke-free months later. PMID- 16340516 TI - Concurrent quit & win and nicotine replacement therapy voucher giveaway programs: participant characteristics and predictors of smoking abstinence. AB - This study compares the participant characteristics, program costs, and outcomes of a Quit & Win contest and a nicotine replacement therapy (NRT) voucher giveaway promotion. Both programs were conducted simultaneously so that smokers could enroll in either one program alone (n = 849 and 690, respectively) or both programs (Combination group; n = 230). A follow-up telephone survey of a random sample of participants was conducted 4 to 7 months after enrollment to evaluate smoking status. At enrollment, participants in the three groups were comparable on most smoking and demographic variables, although Quit & Win participants were, on average, younger than those who signed up to get the NRT voucher. Compared with the characteristics of smokers in the region, those who enrolled in the intervention programs were heavier smokers and had more years of formal education. At follow-up, the self-reported quit rates were similar across the three intervention groups, ranging between 25 percent and 30 percent. The only evidence for a higher quit rate among those in the Combination group was among younger smokers. On a simple estimated cost per quit basis, the Quit & Win (130 dollars) and NRT (179 dollars) voucher interventions appear roughly comparable. In all groups, abstinence rates were higher among lighter smokers (<21 cigarettes per day), participants who did not live with another smoker, and those who were married. Both the Quit & Win and NRT voucher giveaway programs were effective in recruiting smokers to make a quit attempt, although combining both interventions did not generally increase abstinence rates. PMID- 16340517 TI - Giving away free nicotine medications and a cigarette substitute (Better Quit) to promote calls to a quitline. AB - This article presents results from two population-based promotions for free cessation products used to induce smokers to call the New York State Smokers'. The first promotion was a press announcement urging smokers to call the quitline to get a voucher for a free 2-week supply of nicotine patches or gum (nicotine replacement therapy [NRT]). The second promotion involved comparing response to two newspaper advertisements for the quitline, one of which offered a free stop smoking guide and one that offered the guide plus a free stop smoking aide called Better Quit (BQ). Responses to each promotion were monitored by tracking the call volume to the quitline-before, during, and after each of the promotions. The NRT voucher promotion increased median call volume 25-fold compared to prepromotion levels, whereas the BQ newspaper advertisement increased median call volume 2 fold compared to a newspaper advertisement for the quitline, which did not offer BQ. A random sample of 732 smokers who received the free NRT voucher were followed up 4 to 6 months later to determine their use of NRT and smoking behavior. Embedded within this follow-up study was a randomized experiment in which half the smokers were mailed a quit kit with BQ included and the other half received just the quit kit. Seventy percent of follow-up survey respondents said that they had redeemed the NRT voucher and used the medication to try to quit smoking. Twenty-two percent reported they were no longer smoking, compared with the 12 percent among a comparison group of quitline callers who had not received the free NRT voucher (odds ratio = 1.77; 95% confidence interval: 1.17-2.68). There was no difference in quit rates between those that were sent the BQ cigarette substitute and those that did not get the BQ. This study shows that offering a free 2-week voucher for NRT is a cost-effective method to increase calls to a stop smoking quitline and may also increase the odds of quitting for those who get the free NRT. PMID- 16340518 TI - Smoking in pregnancy: patient and provider risk reduction behavior. AB - OBJECTIVE: To describe the range of risk reduction behaviors among women who continue to smoke after learning of their pregnancy, including reduced tobacco use, eventual cessation, and sustained abstinence as well as the patient-reported smoking cessation-promoting behaviors of prenatal care providers. METHODS: This research is part of a larger prospective, community-based study conducted to assess the association between maternal stress and birth outcomes and infant health and development. Over a 2-year period, from February 2000 to November 2001, women receiving prenatal care at a consortium of public health centers in Philadelphia, Pennsylvania, were consecutively recruited (n = 1,451) completing interviews at their initial prenatal visit and again 3 to 4 months following their delivery. Smoking rates during pregnancy were determined from responses given during the first postpartum interview, at 3 to 4 months postpartum. RESULTS: Of the 1,451 women interviewed at 3 to 4 months postpartum, 24.9 percent indicated smoking during their pregnancy. Of these antenatal smokers, 89.0 percent reported reducing their cigarette consumption during pregnancy. However, only 25.4 percent attained abstinence during their pregnancy. Among women who achieved abstinence during their pregnancy, 21.7 percent were still not smoking at the time of the postpartum interview. Antenatal smokers reported that prenatal care providers asked about their smoking (90.6%) and advised about quitting (76.5%). However, only 27.9 percent were given referrals to smoking cessation programs. CONCLUSION: While cessation was achieved by only a quarter of antenatal smokers, almost 90 percent reduced their cigarette consumption. Prenatal care providers identified and provided cessation advice to the majority of women who were smoking but they did not follow through with material assistance in the form of referrals to smoking cessation programs. PMID- 16340519 TI - Status of quality improvement activities to improve immunization practices and delivery: findings from the immunization quality improvement symposium, October 2003. AB - The Centers for Disease Control and Prevention convened a symposium on 22-23 October 2003 to bring together investigators and stakeholders working to apply the quality improvement (QI) approaches to immunization delivery in individual medical practices. The goal was to identify effective program components and further development of model programs. A call for projects was widely disseminated; of 61 submissions received, eight projects were selected. Three of the eight programs used the "train the trainer" approach, three used site specific training, one used a "practice collaborative" approach, and one employed the use of tracking and outreach workers to effect change. At the symposium, invited experts reviewed each program. Common program features that appeared effective included involvement of a variety of staff within the office environment, collection and review of site-specific performance measurements to identify gaps in delivery, periodic monitoring of performance measurement to revise interventions and maintain the improvements, and provision of formal continuing education credits. While research is needed on ways to promote and integrate QI into practices, it is likely that a variety of QI strategies will be shown to be effective, depending on the clinical settings. The field will benefit from standardized outcome measures, cost analysis, and evaluation, so comparisons can be made among different programs. PMID- 16340520 TI - The value of microgrants for community-based health promotion: two models for practice and policy. AB - In 2001, the Office of Disease Prevention and Health Promotion in the US Department of Health and Human Services announced its intention to (1) identify innovative ways to increase public awareness and focus on Healthy People 2010 objectives and (2) broaden the participation of community-based organizations, including agencies new to public health. The mechanism selected, microfinancing, was modeled after small venture loans for economic stimulus in developing countries. The Office of Disease Prevention and Health Promotion selected one state health department and one academic research organization from 80 applicants to test models of awarding "microgrants" of 2,010 dollars to community agencies. This article describes the two models, the types of agencies that were funded, the primary Healthy People 2010 objectives targeted, examples of how the monies were used and leveraged by grantees, and the implications of microgrants for public health practice and policy. PMID- 16340521 TI - Hurricane Isabel-related mortality--virginia, 2003. AB - BACKGROUND: Hurricane Isabel had a massive negative environmental, public health, and economic impact; Virginia bore the highest death toll (32) among nine states affected by this storm. A descriptive mortality analysis was conducted to identify modifiable risk factors and corresponding injury prevention measures that might mitigate future natural disaster-related morbidity and mortality in Virginia. METHODS: Information for the decedents, including demographic data, health status, and injury circumstances, was collected from the records of the Virginia Office of the Chief Medical Examiner and Office of Vital Records/Health Statistics. Criteria from the National Hurricane Center were used to classify deaths as direct or indirect. Storm assessments and emergency-response reports were also reviewed. RESULTS: A total of 32 deaths associated with Hurricane Isabel occurred in several densely populated localities in southeastern and central Virginia. The median age of decedents was 48 years (range: 7-85 years). A disproportionately higher mortality (21 [66%] of 32) occurred among persons older than 45 years (Virginia 2000 Census data). Twelve deaths were directly caused by environmental factors related to the storm (eg, seven drowning deaths and five traumatic head injuries from falling trees). Twenty deaths were indirectly associated with the storm and its effects: six fatal motor vehicle crashes, five related to clean-up operations, seven associated with power outages, and two stress-related (ie, myocardial infarction and suicide). The presence of alcohol or drugs was observed in 9 (28%) of 32 deaths. CONCLUSIONS: Classifying deaths as direct or indirect facilitates better target interventions on the basis of the identification of modifiable risk factors underlying hurricane-associated fatal injuries. Public education messages that reinforce avoidance of use of alcohol and drugs during natural disaster situations might reduce risk for injury. PMID- 16340522 TI - Meetings, meetings, and more meetings. AB - The Management Moment" is a regular column within the Journal of Public Health Management and Practice. Janet Porter, PhD, and Edward L. Baker, MSc, MD, MPH, are serving as The Management Moment Editors. Dr Porter is Associate Dean for Executive Education, The North Carolina Institute for Public Health, School of Public Health, at the University of North Carolina at Chapel Hill, and Dr Baker is Director of The North Carolina Institute for Public Health, School of Public Health, at the University of North Carolina at Chapel Hill. This column provides commentary and guidance on timely management issues commonly encountered in public health practice. PMID- 16340523 TI - Primary pars plana vitrectomy versus scleral buckle surgery for the treatment of pseudophakic retinal detachment: a randomized clinical trial. AB - OBJECTIVE: To compare the anatomical and functional outcome of scleral buckle (SB) surgery with that of pars plana vitrectomy (PPV) alone in the treatment of primary rhegmatogenous pseudophakic retinal detachment (RD). METHODS: In this prospective, randomized clinical trial, 150 eyes of 150 patients with pseudophakic RD and proliferative vitreoretinopathy (PVR) stage B or less were randomized to SB surgery (75 eyes) or primary PPV (75 eyes). SB surgery involved break localization, cryotherapy, placement of a circumferential 240 style 2.5-mm solid silicone band, combined with a local buckle when indicated, and transscleral drainage of subretinal fluid. PPV included extensive vitreous removal, perfluoro-n-octane injection or endodrainage of subretinal fluid to flatten the retina, cryopexy treatment of breaks, and fluid/air exchange with injection of 20% SF6. Postoperative follow-up was 1 year. Break diagnosis, operating time, intraoperative and postoperative complications, retinal reattachment rate for single as well as multiple surgeries, axial length changes, and best-corrected visual acuity at 1 year after surgery were the main outcome measures. RESULTS: The number of eyes that were diagnosed with additional breaks inter-operatively was higher in the PPV group (P=0.004, chi test). Mean operating time was significantly less (P=0.0001, t-test) in the PPV group. With a single surgery, the retina was reattached in 62 eyes (83%) in the SB surgery group and in 71 eyes (94%) in the PPV group (P=0.037, Fisher exact test). With subsequent surgeries, final anatomical reattachment was achieved in 71 cases in the SB surgery group and in 74 cases in the PPV group (P=0.37, Fisher exact test). Mean axial length change at 1 year was 0.95 mm in the SB surgery group and 0.1 mm in the PPV group (P=0.0001, t-test). Mean final best-corrected visual acuity (logMAR) was 0.40 in the SB surgery group and 0.33 in the PPV group (P=0.26, t test). CONCLUSIONS: Primary PPV offers potential advantages over SB surgery in the treatment of pseudophakic RD, including less operating time, accurate diagnosis of breaks, higher reattachment rate with a single surgery, and no postoperative axial length changes. Retinal reattachment rate with multiple surgeries and final visual acuity at 1 year were similar for SB surgery and PPV. PMID- 16340524 TI - 10-year follow-up of visual functions in patients who underwent scleral buckling. AB - PURPOSE: To study long-term visual functions in eyes that underwent scleral buckling for treatment of rhegmatogenous retinal detachment. METHODS: We retrospectively studied a nonconsecutive case series of 205 eyes with retinal detachment that underwent scleral buckling and were followed up for 10 years after surgery. Preoperatively, postoperatively, and 10 years after surgery, best corrected visual acuities and visual fields were recorded. RESULTS: During the first postoperative year and during the next 9 years, there were significant increases in visual acuity (P<0.0001 and P<0.0001, respectively). Between the operation and 1 month postoperatively, there was a significant increase in the area size of visual field measured by Goldmann perimetry (V-4 isopter, P=0.0001; I-4 isopter, P=0.0001). Between 1 month and 10 years after surgery, there was no significant change of the area size of each isopter (V-4, P=0.786; I-4, P=0.211). CONCLUSION: Improvement in visual acuity after scleral buckling is possible over a long period. There may be a limit to long-term improvement of visual field after scleral buckling. PMID- 16340525 TI - Triamcinolone-assisted internal limiting membrane peeling. AB - PURPOSE: To review our experience with triamcinolone-assisted pars plana vitrectomy for internal limiting membrane (ILM) peeling for various retinal diseases. METHODS: Twenty-one patients underwent surgery in which intraoperative triamcinolone acetonide (TA) was used. Indications for surgery included epiretinal membrane (3 patients), branch retinal vein occlusion associated with macular edema (2), traction retinal detachment (3), diabetic macular edema (4), vitreous hemorrhage with diabetic macular edema (4), macular hole (4), and cystoid macular edema (1). RESULTS: TA was useful in the removal of the ILM in all cases. There were no intraoperative complications or toxicity. The mean follow-up was 22 weeks (range, 9-30 weeks). Eleven patients improved by >or=2 Snellen lines, 1 lost >or=2 Snellen lines, and 9 were within 2 Snellen lines of preoperative vision at the last follow-up. CONCLUSIONS: The intraoperative use of TA improves visualization of ILMs associated with a variety of conditions. No intraoperative or postoperative complications were observed. TA-assisted removal of the ILM appears to be safe and cost effective. TA-assisted ILM peeling should be considered as an alternative to the use of intraoperative dyes. PMID- 16340526 TI - Managing severe endophthalmitis with the use of an endoscope. AB - BACKGROUND: To demonstrate the value of ophthalmic endoscopy in treating patients with severe vision-threatening endophthalmitis in whom visualization through the anterior ocular structures is compromised. METHOD: In this retrospective study of patients with endophthalmitis operated on with the use of an endoscope between 1997 and 2003, demographics, cause of endophthalmitis, preoperative and postoperative vision, comorbid factors, complications, intraoperative retinal appearance, etiology, and additional procedures required were recorded. RESULTS: Fifteen patients were included. A variety of causes were observed: endogenous endophthalmitis (n=3), penetrating trauma (n=2), post-cataract extraction (n=7), bleb infections (n=2), and post-retinal detachment surgery (n=1). Six-month follow-up is reported. Positive cultures were obtained in all but one case. Several patients had aggressive pathogenic organisms. Eight patients retained useful vision. Six of eight patients without retinal necrosis by endoscopic examination had improved vision, with final visual acuity varying between counting fingers and 20/20. Only 1 of 6 patients with retinal necrosis retained useful vision at 20/50. CONCLUSION: The ophthalmic endoscope aids in performing safe, diagnostic, and therapeutic vitrectomy in endophthalmitis. Prognosis is dependent on preoperative visual acuity and retinal appearance seen at the time of endoscopic surgery. PMID- 16340527 TI - Effect of subthreshold infrared laser treatment for drusen regression on macular autofluorescence in patients with age-related macular degeneration. AB - BACKGROUND: Subthreshold laser therapy has been shown to cause drusen reduction. Using this method, visible laser burns are not created in the retina; presumably, the energy is absorbed by the retinal pigment epithelium (RPE) and stimulates reabsorption of drusen material, sparing photoreceptors from destruction. We hypothesized that autofluorescence (AF) changes would be visible after such treatment and might be sensitive to quantify RPE changes. METHODS: Twenty eyes of 13 patients with non-exudative age-related macular degeneration were studied. Forty-eight subthreshold infrared diode laser spots were applied as treatment to cause drusen regression. Before, immediately after, and 3 months after treatment, color fundus photography, fluorescein angiography (FA), and autofluorescence (AF) imaging were performed. Treated lesions were identified on overlay images of all three imaging methods. RESULTS: : The averaged sensitivity of AF imaging compared with FA was 29.6+/-28.7% versus 10.2+/-12.2% (P=0.008) at the immediate posttreatment time point and 43.5+/-28.7% versus 33.8+/-26.5% (P=0.043) at the 3 month posttreatment time point, respectively. Reduction of drusen at 3 months correlated with the detection sensitivity of AF imaging at the immediate posttreatment time point (P=0.022). CONCLUSION: Immediately after subthreshold laser treatment, AF imaging was more sensitive to detect RPE changes than FA. This suggests that noninvasive AF imaging may allow prediction of the effect of subthreshold laser treatment and might be used to titrate treatment dose. PMID- 16340528 TI - Acute central serous chorioretinopathy and fundus autofluorescence. AB - OBJECTIVES: To describe fundus autofluorescence (FAF) in a series of patients with acute central serous chorioretinopathy (CSC). METHODS: Nine eyes of six patients with acute CSC were evaluated with fluorescein angiography (FA) and FAF imaging to evaluate the nature of the focal retinal pigment epithelial (RPE) leak evident with FA. RESULTS: All nine eyes in this series demonstrated hypo autofluorescence corresponding precisely to the site of the focal RPE leak seen on FA. CONCLUSIONS: In this group of patients, the acute focal RPE leaks seen with FA corresponded precisely to an area of hypo-autofluorescence imaged with FAF. This observation supports the concept that a mechanical defect or absence of the RPE accounts for the leakage from the inner choroid to the sub-neurosensory space in CSC. FAF is also a useful noninvasive diagnostic adjunct to identify the focal RPE leak in patients with acute CSC. PMID- 16340529 TI - Lidocaine gel anesthesia for intravitreal drug administration. AB - PURPOSE: To describe the use of topical gel anesthesia for intravitreal injection drug delivery. METHODS: The first group (n=12) of patients is a crossover group of patients who on separate occasions received intravitreal injections after both 2% lidocaine subconjunctival injection (SC) and 2% lidocaine topical gel anesthesia using 30-gauge needle. The second group (n=16) is a consecutive group of patients who received either SC (n=8) or gel (n=8) anesthesia before intravitreal injection of triamcinolone using 27.5-gauge needle. Descriptive and numerical pain analog scale was used to assess pain sensation. RESULTS: : There was no difference in pain score between gel and SC within the first group (P=0.67, paired, nonparametric test) and no difference in pain score comparing patients who had either SC or gel anesthesia before 27.5-gauge intravitreal injections (P=0.82, unpaired t-test) in the second group. However, there were significant differences in incidence of chemosis (P<0.001) and subconjunctival hemorrhage (P<0.001) after injection versus gel anesthesia in both groups. CONCLUSION: Anesthesia with lidocaine 2% gel provides satisfactory patient comfort for administration of intravitreal injection and causes less chemosis and hemorrhage than SC anesthesia. PMID- 16340530 TI - Autosomal dominant pattern dystrophy: identification of a novel splice site mutation in the peripherin/RDS gene. AB - PURPOSE: To describe the clinical features of and identify the mutation responsible for an autosomal dominant pattern dystrophy occurring in a three generation family. METHODS: Five affected family members underwent clinical examination and additional testing including intravenous fluorescein angiography where indicated. Mutation screening of the peripherin/RDS gene was performed. RESULTS: Visual acuity ranged from 20/20 to counting fingers. All patients who reported vision loss noted the onset after the age of 40 years. Predominantly perifoveal, discrete, retinal pigment epithelial changes were present in all patients. Two patients had extensive peripheral yellowish flecks, and one had an atrophic macular scar. Mutation screening of the complete peripherin/RDS coding sequence and exon/intron boundaries revealed a novel splice site mutation. CONCLUSION: A three-generation family with an autosomal dominant pattern dystrophy arising from a previously unreported splice site mutation in the RDS gene is described. PMID- 16340531 TI - Prevalence, clinical features, and causes of vision loss among patients with ocular toxocariasis. AB - BACKGROUND: To describe the prevalence, clinical features, and causes of vision loss among patients with ocular toxocariasis seen at a uveitis referral center. METHODS: A review was completed of the charts of patients with ocular toxocariasis who were examined between 1977 and 1996 at the Francis I. Proctor Foundation of the University of California at San Francisco. The prevalence of ocular toxocariasis among all uveitis patients seen at the center was determined. Demographic features, symptoms, and signs in all patients were evaluated. RESULTS: Ocular toxocariasis occurred in 22 (1.0%) of 2,185 uveitis patients. The mean patient age was 16.5 years. Inflammation was usually unilateral (90.9%). Toxocara uveitis presented as a granuloma in the peripheral retina in 50% of cases, as a granuloma in the macula in 25% of cases, and as a moderate to severe vitreous inflammation mimicking endophthalmitis in 25% cases. The primary causes of vision loss were vitritis (52.6%), cystoid macular edema (47.4%), and traction retinal detachment (36.8%). CONCLUSIONS: Ocular toxocariasis is an uncommon cause of uveitis that mainly affects younger patients. Inflammation is typically unilateral and presents as either a granuloma in the peripheral or posterior retina or a moderate to severe vitreous inflammation mimicking endophthalmitis. PMID- 16340532 TI - Short-term inter-visit variability of erg amplitudes in normal subjects and patients with retinitis pigmentosa. AB - PURPOSE: To evaluate the short-term test/retest variability in visually normal subjects and patients with retinitis pigmentosa (RP), and to assess the effect of stimulus intensity and baseline amplitude on electroretinogram (ERG) variability. METHODS: Eighteen patients with RP and nine visually normal subjects had a series of three unilateral ERGs, with an inter-visit interval of no less than 2 days and no more than 2 weeks. Responses to dark-adapted and both light-adapted single flash and 32 Hz flicker stimuli were recorded from a dilated eye over a range of stimulus intensities. B-wave amplitudes were compared to baseline amplitudes recorded at initial visit, and the resulting inter-visit percent difference was compared between stimulus intensities. Inter-visit variability was determined by one-way repeated measures analysis of variance using a 95% confidence interval to calculate threshold criteria for significant change. Analysis of variance followed by Bonferroni test for pairwise comparison was used to test for differences in inter-visit variability between two RP patient subgroups (higher versus lower baseline amplitudes) and visually normal subjects. The effect of stimulus intensity on amplitude reproducibility was also assessed. RESULTS: Threshold for significant increase or decrease in inter-visit ERG amplitudes at a 95% confidence level for patients with RP and visually normal subjects was often at or above 25% and not infrequently at or above 40% for certain stimuli and test conditions. While no statistical difference in inter-visit variability was demonstrated between visually normal subjects and patients with RP who were arbitrarily categorized as having relatively higher baseline amplitudes, there was a difference between each of these two groups and a smaller group of patients with RP categorized as having lower baseline amplitudes. Although the authors could not demonstrate that percent inter-visit differences varied with stimulus intensity in either controls or patients with RP, the 32 Hz flicker stimulus generally produced less amplitude variability than either dark- or light-adapted single flash stimuli in patients with RP. CONCLUSIONS: When using ERG amplitude for monitoring either the natural history of functional loss in retinal disease or as an outcome measure during a therapeutic trial, it becomes vital to define inter-visit variability of ERG amplitudes. These findings suggest that a percentage of patients with RP with appreciably lower baseline ERG amplitudes may manifest greater inter-visit ERG amplitude variability than patients with RP with higher baseline amplitudes or controls. Stimulus intensity had no clinically significant effect on inter-visit amplitude variability. PMID- 16340533 TI - Effective transscleral delivery of two retinal anti-angiogenic molecules: carboxyamido-triazole (CAI) and 2-methoxyestradiol (2ME2). AB - PURPOSE: To evaluate the human transscleral diffusion and intravitreal delivery of carboxyamido-triazole (CAI) and 2-Methoxyestradiol (2ME2). METHODS: The transscleral diffusion of two retinal antiangiogenic molecules, CAI and 2ME2, was measured in vitro to assess their potential transscleral delivery. Varying concentrations and different solvents of CAI and 2ME2 were placed in the upper compartment of a two-chamber acrylic perfusion apparatus, on the episcleral side of the sclera obtained from human donor eyes. Samples were taken from the lower compartment (uveal side) for up to 24 hours and measured by high performance liquid chromatography. RESULTS: All three solutions that contained CAI efficiently diffused through the sclera with permeability constants that ranged from 2.8 to 5.5 x 10 cm/s. The scleral permeability constant derived for 2ME2 was 9.96 x 10 cm/s. The permeability constants obtained for both CAI and 2ME2 are similar to each other as well as to permeability constants measured for other small molecules such as fluorescein and dexamethasone fluorescein. CONCLUSION: Both CAI and 2ME2 traverse the sclera efficiently. These data combined with the reported inhibition of posterior segment neovascularization observed with these two molecules demonstrates that CAI and 2ME2 are good candidate molecules to treat posterior segment neovascularization by local delivery. PMID- 16340534 TI - Histologic analysis of bioabsorbable scleral buckling implants: an experimental study on rabbits. AB - PURPOSE: To analyze histologically tissue reactions to bioabsorbable PLA96 in rabbit eyes. METHODS: Scleral buckling operations were carried out in 48 rabbits. Two materials were used: bioabsorbable PLA96 (polylactide 96/4; L/D molar ratio 96/4) and silicone sponge. One eye of each rabbit was operated on and the other eye served as a nonoperated control. After follow-up times of 1, 3, 5, and 12 months, the rabbits were killed and the eyes enucleated for histology. RESULTS: All rabbits recovered well. Histologically, tissue reactions were very localized; implant fragments were not seen within the sclera. The amounts of fibrous tissue and inflammatory cells (mainly macrophages) inside the implant area increased over time. One rabbit from the silicone group was killed 4 months postoperatively owing to refusal to eat. In the PLA96 group, acute or chronic infections occurred in four rabbits. The bioabsorbable implant was macroscopically easily detectable at 12 months postoperatively. CONCLUSIONS: The PLA96 material used for scleral buckling in rabbits showed good biocompatibility. The material did not undergo biodegradation during the follow-up period of 12 months. PLA96 implants were associated with thicker fibrous tissue encapsulation and more inflammatory cells compared with silicone sponge implants. PMID- 16340535 TI - Ultrastructural analysis of rabbit retina irradiated with a new 670-nm diode red laser at different powers. AB - PURPOSE: Diode lasers have compact dimensions, efficient electric-optical conversion, absence of major cooling requirements, long useful life, and minimal maintenance. We verified the structural and ultrastructural effects of diode red laser emissions at 670 nm on pigmented rabbit retina. METHODS: Transpupillary laser pulses were applied to rabbit retinas at variable powers with a spot size of 500 microm ( approximately 1,000 microm at the retina level) and duration of 200 milliseconds. Light and electron microscopic analyses were performed. RESULTS: Subthreshold non-visible lesions were produced using a power of 200 mW. Ophthalmoscopically visible lesions were produced using a power of 400 mW. More prominent lesions were obtained using a higher power (600 mW). Structural and peculiar ultrastructural changes correlated with the power used. In general, damage was incurred by the retinal pigment epithelium with extension into the superficial choroid posteriorly and into the outer retina anteriorly. The inner retina was involved at higher power levels. CONCLUSIONS: Ultrastructural analysis of rabbit retina treated with the 670-nm diode laser adds newer insights on tissue changes after thermal irradiation. The effects observed are consistent with those reported for the krypton red laser. The 670-nm diode laser is an economic small-size photocoagulator that appears to be a promising modality for transpupillary laser photocoagulation of the retina. PMID- 16340536 TI - Transpupillary thermotherapy: effect of wavelength on normal primate retina. AB - PURPOSE: To correlate changes in primate fundus after transpupillary thermotherapy (TTT) at two wavelengths. METHODS: Twelve primate eyes were treated with TTT using a wavelength of 635 nm (n=7) or 810 nm (n=5). Laser parameters were as follows: 635 nm (spot size, 1 mm; duration, 30-8 seconds; and fluence [power over time], 20-91.4 J/cm) and 810 nm (spot size, 2 mm; duration, 60 seconds; and fluence, 96-436 J/cm). Fundus photography, fluorescein and indocyanine green angiography, and enucleation were performed at time 0 or 2 weeks after TTT for histologic analysis. RESULTS: Threshold for fundus lesions (91.4 J/cm at 635 nm and 191 J/cm at 810 nm), acute and chronic retinal damage shown by histologic analysis (79.2 J/cm at 635 nm and 96 J/cm at 810 nm), and choroidal vessel occlusion (50 J/cm at 635 nm and 96 J/cm at 810 nm) were lower at 635 nm. Disorganization of the retina and retinal pigment epithelium was seen for both wavelengths at time 0 and 2 weeks after TTT. Occlusion of the choriocapillaris and choroidal stromal vessels was noted only in specimens obtained 2 weeks after TTT. CONCLUSIONS: TTT resulted in acute and delayed damage to the neurosensory retina that persisted at 2 weeks. The 635-nm wavelength demonstrated a lower threshold fluence for visible fundus lesions, retinal damage, and choroidal vascular occlusion than the 810-nm laser. PMID- 16340537 TI - Oral administration of lumiracoxib reduces choroidal neovascular membrane development in the rat laser-trauma model. AB - PURPOSE: To determine whether lumiracoxib, a highly selective cyclooxygenase-2 (COX-2) inhibitor that exhibits anti-inflammatory and antiangiogenic properties, can inhibit experimental choroidal neovascular membrane (CNVM) development induced by focal laser trauma in a well-characterized Brown Norway rat CNVM model. METHODS: Over a 35-day period, 24 rats received daily oral gavage dosages of 20 mg/kg lumiracoxib in a 0.5% (w/v) suspension of sodium carboxymethylcellulose (CMC), while a control group received the 0.5% CMC suspension only. After 7 days, eight laser photocoagulation sites were placed concentrically around the optic disk to induce CNVMs. Thirty-five days later, fundus photography and fluorescein angiography (FA) were performed and eyes were processed for histopathologic analysis. RESULTS: Masked FA grading of lesion sites revealed a small, but statistically significant difference (P<0.0001) in late stage staining intensity and leakage between the mean group scores of treated (1.4) and control (1.7) eyes. Histopathologic analysis demonstrated that the mean CNVM thickness +/- SD of 38 +/-19 microm (n=24 eyes, 175 photocoagulation sites) in the lumiracoxib-treated animals was reduced by 30% (P<0.001) compared to the CNVM mean thickness+/- SD of 54+/- 20 microm (n=24 eyes, 171 photocoagulation sites) in the control animals. CONCLUSION: Systemic administration of the selective COX-2 inhibitor lumiracoxib results in a partial but significant reduction in CNVM development in the rat laser-trauma model and thus may be clinically beneficial as a potential inhibitor of CNVM formation in exudative age-related macular degeneration. PMID- 16340538 TI - neovascular age-related macular degeneration: Natural History and Treatment Outcomes. AB - BACKGROUND: This review summarizes the data reported in peer-reviewed literature and presents current knowledge on differentiation, natural history, and therapeutic outcomes of neovascular age-related macular degeneration (AMD). METHODS: The MEDLINE database was searched to review natural history of neovascular AMD and therapeutic effects of available treatments. RESULTS: The search produced>7,000 articles. Research suggests that fluorescein angiographic characterization of location, composition, and size of neovascular lesions may be important in prognosis and should be considered for evaluation of treatment benefits in conjunction with evidence of recent disease progression for lesions not composed of predominantly classic choroidal neovascularization (CNV). Laser photocoagulation, photodynamic therapy with verteporfin, and administration of pegaptanib sodium reduce the risk of vision loss in selected cases of neovascular AMD, while submacular surgery can reduce the risk of severe visual acuity loss in selected cases of predominantly hemorrhagic CNV; further approaches are under investigation. CONCLUSION: Visual prognosis of neovascular AMD is variable according to lesion location, composition, and size. Often, lesions have a poor prognosis, resulting in rapid and progressive loss of visual acuity and contrast sensitivity. Such losses have a profound effect on patients' quality of life and ability to perform everyday tasks. Reducing the risk of further loss of visual acuity and contrast sensitivity might enable patients with neovascular AMD to maintain better functional abilities. PMID- 16340539 TI - Diagnostic & therapeutic challenges. PMID- 16340540 TI - Bilateral macular hemorrhage associated with autoimmune hemolytic anemia. PMID- 16340541 TI - Endogenous scedosporium apiospermum endophthalmitis. PMID- 16340542 TI - Cancer-associated retinopathy in a patient with hepatocellular carcinoma: case report and literature review. PMID- 16340543 TI - Disappearance of puncta after uveitis in an eye with fundus albipunctatus. PMID- 16340544 TI - Decreased venous tortuosity associated with resolution of macular edema after intravitreal injection of triamcinolone. PMID- 16340545 TI - Flap displacement during vitrectomy 24 months after laser in situ keratomileusis. PMID- 16340546 TI - Transient closure of a retinal capillary hemangioma with verteporfin photodynamic therapy. PMID- 16340547 TI - Dystrophic calcification of the fibrous capsule around a hydrogel explant 13 years after scleral buckling surgery: capsular calcification of a hydrogel explant. PMID- 16340548 TI - A convenient method to concentrate triamcinolone acetonide for intravitreal injection. PMID- 16340549 TI - Photoreceptor atrophy in acute posterior multifocal placoid pigment epitheliopathy demonstrated by optical coherence tomography. PMID- 16340550 TI - Optical coherence tomography in a case of apparent double macular holes. PMID- 16340551 TI - Spontaneous regression of degenerative retinoschisis associated with outer leaf retinal breaks. PMID- 16340552 TI - A new scleral depressor for premature infants. PMID- 16340554 TI - Correspondence. PMID- 16340556 TI - Correspondence. PMID- 16340558 TI - Correspondence. PMID- 16340559 TI - Laparoscopy-assisted total gastrectomy for early gastric cancer: comparison with conventional open total gastrectomy. AB - Laparoscopy-assisted distal gastrectomy has been applied to the treatment of early gastric cancer in Japan. So far, several studies about comparison between laparoscopy-assisted distal gastrectomy and conventional open distal gastrectomy were reported. However, there are few reports on the laparoscopy-assisted total gastrectomy, mainly because this procedure is performed relatively infrequently, and the procedure is more difficult than laparoscopy-assisted distal gastrectomy. This was a case-control study comparing between laparoscopy-assisted total gastrectomy group and open total gastrectomy group. From June 2001 to August 2004, laparoscopy-assisted total gastrectomy was performed in 20 patients. Reconstruction was performed by Roux-en-Y method or Roux-en-Y with jejunal pouch method through the mini-laparotomy. These cases were compared with 19 cases of open total gastrectomy, regarding operating time, blood loss, leukocyte count, C reactive protein, time to the first passage of gas, time to initiate oral intake, and postoperative hospital stay.Laparoscopy-assisted total gastrectomy was successful in 20 patients. The mean operating time was 280 minutes and blood loss was 227.5 mL. Leukocyte counts on days 1, 3, and 7 were significantly lower in laparoscopic surgery group than in open surgery group. The time to first flatus, time to initiate oral intake, and postoperative hospital stay was significantly shorter (P < 0.05) in the laparoscopic surgery group than in the open surgery group. This study demonstrated that laparoscopy-assisted total gastrectomy is suitable and feasible for early gastric cancer and has the advantage of a shorter recovery time compared with open total gastrectomy. PMID- 16340560 TI - Management of acute calculous cholecystitis in high-risk patients: percutaneous cholecystotomy followed by early laparoscopic cholecystectomy. AB - Emergency cholecystectomy for acute cholecystitis is associated with high morbidity and mortality rates in patients with significant comorbidities and high risk surgery. The aim of this study was to evaluate the effectiveness, possible advantages, and complications of percutaneous cholecystostomy (PC) followed by an early laparoscopic cholecystectomy (LC) in relation to conservative treatment followed by a delayed LC in high-surgical risk patients. Between 2002 and 2004, patients were randomly classified into 2 groups: the first group consisted of patients who had PC followed by an early LC (PCLC group, n = 31) and the second group consisted of patients who had conservative treatment followed by a delayed LC (DLC group, n = 30). The groups were statistically compared regarding their demographic, comorbidity, hospital stay, conversion, and complication rates. PC was technically successful in 31 patients with no attributable mortality or major complications. No difference had been found in regarding demographic, comorbidity, and complication rates. In PCLC group, all the patients experienced symptom relief within 24 hours, and early LC was attempted in 31 patients once their clinical condition was sufficiently stable, this was successfully accomplished in 29 (93.5%). In the DLC group, delayed LC was attempted in 30 patients, and this was successfully accomplished in 26 (86.6%). The hospital stay was shorter and cost was in the PCLC group was lower than in the DLC group. PC allows resolution of sepsis in patients at high surgical risk. Early LC could be safely performed once sepsis and acute infection resolved in these patients. PMID- 16340561 TI - Laparoscopy-assisted ileal pouch-anal anastomosis: surgical outcomes after 10 cases. AB - We analyzed outcomes of laparoscopy-assisted ileal pouch-anal anastomosis (LAIPA) from 10 patients (7 with ulcerative colitis and 3 with familial adenomatous polyposis) operated on between January 1998 and March 2004. Median operating time was 246 minutes. There were no intraoperative complications. There were no conversions. Postoperative complications occurred in 3 (30%) patients: 2 cases of wound infection and 1 case of a foreign body retrieved during pouch endoscopy. There was a 30% reoperation rate due to unsuspected duodenal perforation, a persistent postoperative pain, and 1 case of intestinal obstruction after ileostomy closure. There were no deaths. Median time to resumption of diet was 24 hours. Median hospital stay was 7 days. All ileostomies were closed 6 to 8 weeks after LAIPA. LAIPA is feasible and safe and should be selectively offered to nonobese patients. PMID- 16340562 TI - Left upper quadrant entry during gynecologic laparoscopy. AB - The use of the left upper quadrant technique in establishing a pneumoperitoneum during gynecologic laparoscopy was reviewed retrospectively in 267 patients who underwent gynecologic laparoscopy. The study population included all patients presenting to the University of Louisville Hospital outpatient surgery unit for laparoscopic gynecologic surgery from January 1994 to March 2002. Data sheets were prospectively compiled for each patient that included their demographics as well as the intraoperative insufflation technique used and the number of attempts necessary to achieve successful insufflation. All recognized complications associated with establishment of the pneumoperitoneum or insertion of the primary trocar were recorded. The database included 3314 patients of which 267 underwent insufflation via the left upper quadrant technique. Failure to achieve insufflation using this technique occurred in 4 patients (1.5%). There were no bowel or vessel injuries. Puncture of the left lobe of the liver occurred in 3 patients (1.12%). All injuries were managed without laparotomy, and the patients fully recovered without sequelae. The left upper quadrant entry technique is an effective means of establishing a pneumoperitoneum in patients undergoing gynecologic laparoscopic procedures. PMID- 16340563 TI - Impedance cardiography: noninvasive measurement of hemodynamics and thoracic fluid content during endoscopic thoracic sympathectomy. AB - Endoscopic thoracic sympathectomy (ETS) is a minimally invasive procedure for treating palmar hyperhidrosis (PH). Hemodynamic changes are associated with CO(2) insufflation during ETS. In the present study, we examined hemodynamic changes during general anesthesia using impedance cardiography (ICG) monitor. Seventeen adult patients (15 males) scheduled to undergo elective unilateral ETS for treatment of PH were enrolled in the study. Patients with cardiorespiratory diseases were excluded from the study. Their age and weight mean values were 26.5 +/- 5 years, 71.9 +/- 11.5 kg, respectively. Besides routine monitoring, impedance cardiography monitor was used to measure cardiac output (CO), cardiac index (CI), stroke volume (SV), thoracic fluid content (TFC), and systemic vascular resistance (SVR). Three phases were defined for data collection: A, prior to CO(2) insufflation; B during gas insufflation (at 10, 5, and 2 mm Hg intrathoracic pressures); and C, after gas deflation. Repeated-measures analysis of variance (ANOVA) was used for statistical analysis and post hoc Bonferroni test for multiple comparisons of the data obtained. For all comparisons, P < 0.05 was considered significant. Systemic vascular resistance significantly increased at stages B10 and 5 compared with stage A mean value (P < 0.05). CO, CI, and SV mean values decreased significantly at stage B compared with stage A mean values. The mean values of thoracic fluid content at stages A, B10, 5, 2, and C were 33 +/- 5, 30.6 +/- 3.5, 31 +/- 3.4, 31.6 +/- 3.3, and 32.5 +/- 6.8/kOmega, respectively with significant differences (P < 0.05). Significant reductions of cardiac parameters were reported in the present study, but they were of minimal clinical significance. Of interest was the significant reduction of thoracic fluid content during CO(2) insufflation, whether it correlates to the magnitude of compression, caused by CO(2) insufflation accompanied by high systemic vascular resistance or sympathectomy procedure, yet to be further studied. PMID- 16340564 TI - Esophageal Doppler (ODM II) improves intraoperative hemodynamic monitoring during laparoscopic surgery. AB - Minimally invasive laparoscopic surgery has been expanded to the elderly and high risk surgical patients with underlying cardiac and pulmonary disease. However, possible cardiovascular changes during CO2 pneumoperitoneum necessitate close intraoperative monitoring. In this prospective study, 55 patients (mean age 62.52 years, range 26-82) undergoing laparoscopic surgery were included. Patients were categorized into 3 groups of low (group A: 12 patients, mean age 55.5 years), moderate (group B: 22 patients, mean age 59.5 years), and high (group C: 21 patients, mean age 69.71 years) surgical risk according to ASA physical status classification. Similar anesthetic agents and anesthetic techniques were used in the above cases. An esophageal Doppler (ODM II, Abbott Laboratories) was used to measure aortic blood flow velocity and thereby estimating stroke volume (SVe) and cardiac output (COe) throughout anesthesia, in addition to traditional monitoring. After abdominal insufflation (peak intra-abdominal pressure: 13-15 mm Hg) COe values decreased from the initial value after induction of anesthesia by 22%, 20%, and 18% for groups A, B, and C, respectively (P < 0.05). The above values further deteriorated (25%, 28%, and 30% for groups A, B, and C, respectively) in the anti-Trendelenburg positioning of the patient. The peak aortic blood flow velocity (PV) followed the changes, thus indicating that heart muscle contractility is affected during the procedure. Stabilization of the above values was achieved after 20 minutes of CO(2) pneumoperitoneum and improvement was noted only after deflation of the abdomen. Heart rate and blood pressure essentially remained unchanged throughout the procedure, although the final values were increased compared with initial. Insufflation of the abdomen with CO(2) produces measurable effects on the cardiovascular system that require reappraisal of hemodynamic monitoring during anesthesia. ODM II offers a reliable, relatively noninvasive, cost-effective tool for intraoperative monitoring of the hemodynamic changes with a potential for future application for improvement of intraoperative hemodynamic status of patients. PMID- 16340565 TI - Videoscopic thyroidectomy with fine needle-type apparatuses: an approach that does not leave a scar on the neck or anterior chest. AB - A total videoscopic operation with constant CO(2) gas insufflation is ideal for performing a scarless thyroidectomy because the skin incisions at the surgical ports are made far from the neck in the axillar area. However, there are many difficulties associated with the performance of a videoscopic thyroidectomy when approaching pneumo-subcutaneously from distant ports, because the instruments are inserted tangentially toward the thyroid gland from a distant port and their range of manipulation is limited. The devices and surgical techniques used for a videoscopic thyroidectomy that does not leave a scar on the anterior chest or neck were developed. Fine needle-type cautery and forceps that could access directly to the thyroid gland from the neck were used to support the forceps. A CO(2) reservoir bag connected to the CO(2) insufflation tube contributed to the safer manipulation by the creation of a stable operation space during emitting smoke from coagulation device. A silicon ring stopper was useful to prevent dislodging of the ports. In conclusion, videoscopic neck surgery would be easier using these new devices and techniques and would produce better cosmetic results than video-assisted and conventional neck surgeries. PMID- 16340566 TI - A unique case of pancreatitis and retention cyst in esophageal heterotopic pancreas. AB - Heterotopic pancreas is normal pancreatic tissue located in an abnormal location. Heterotopic pancreas in the esophagus is a rare congenital anomaly. We present a unique case of pancreatitis and retention cyst in a heterotopic pancreas in the esophagus, outline the investigation and management of cystic lesions of the distal esophagus, and review the literature. PMID- 16340567 TI - A case of sporadic gastric carcinoid tumor treated successfully by laparoscopy assisted distal gastrectomy. AB - A 49-year-old man was referred to us for treatment of a gastric carcinoid tumor. Gastroscopy revealed a superficial elevated lesion with a central depression covered with nonspecific gastric mucosa located in the anterior wall of the gastric body. The lesion was diagnosed on biopsy as a gastric carcinoid tumor. Preoperative ultrasound and computed tomography examinations revealed a tumor confined to the gastric submucosa and without lymph node metastasis. Therefore, laparoscopy-assisted distal gastrectomy (LADG) was performed. Macroscopically, the resected specimen contained an elevated lesion measuring 0.9 x 0.8-cm with a central depression. The postoperative course was uneventful, and the patient remains free of recurrence 10 months after surgery. There are few cases of sporadic gastric carcinoid tumor successfully treated by LADG. LADG may be useful for treatment of patients with sporadic gastric carcinoid tumor and possible lymph node metastasis. PMID- 16340568 TI - Endoscopic transgastric drainage of pancreatic pseudocyst with the use of Nd:YAG laser. AB - Pancreatic pseudocysts have been successfully managed with endoscopic drainage recently. This report describes a case of endoscopic transgastric drainage using endoscopic ultrasonography (EUS) and an Nd:YAG laser. EUS was used to detect an optimal puncture site of the pseudocyst and to reduce the risk of bleeding and perforation. An Nd:YAG laser was used to minimize the risk of bleeding and to penetrate the thick wall of the pseudocyst. After transgastric cystgastrostomy was performed, an internal stent was placed between the pseudocyst and the stomach. There were no complications associated with endoscopic interventions. Complete resolution of the pseudocyst was observed. Endoscopic transgastric drainage of pancreatic pseudocysts is a recommended approach for selected patients with pancreatic pseudocysts that are uncomplicated and are located adjacent to the stomach. Safe and effective drainage can be achieved without hemorrhage and perforation with the use of EUS, an Nd:YAG laser, and a stent. Furthermore, the Nd:YAG laser facilitated passage through a markedly indurated pseudocyst wall and it seemed to be an effective instrument, especially for pseudocysts with a thick wall. PMID- 16340569 TI - Laparoscopic cholecystectomy for a double gallbladder of the duodenal type. AB - Double gallbladder is a rare congenital malformation and generally considered a duplication of 1 primordium. We encountered an extremely rare case of double gallbladder of the duodenal type that was considered a duplication of 2 primordia. We were able to diagnose the anomaly preoperatively by endoscopic retrograde cholangiopancreatography and spiral computed tomography after intravenous infusion cholangiography, and laparoscopic removal was successfully performed. To our knowledge, this is the first reported case of double gallbladder of the duodenal type that was diagnosed preoperatively and treated successfully by laparoscopic removal. PMID- 16340570 TI - Laparoscopic treatment of liver hemangioma. AB - Cavernous hemangioma is the most common benign liver tumor. When it becomes symptomatic, enucleation is considered the treatment of choice because of its lower morbidity compared with liver resection. Therefore, although there have been many series of laparoscopic liver resection, only a few cases of laparoscopic enucleation of liver hemangiomas have been reported. We report the case of a 36-year-old woman with a symptomatic 4-cm liver hemangioma of the left lobe who underwent laparoscopic enucleation, with complete relief of the symptoms at the 3-month follow-up. The operating time was 90 minutes. Neither liver mobilization nor ligament division was necessary. The dissection was carried out with minimal blood loss. The postoperative period was uneventful, and the patient was discharged on postoperative day 4. Laparoscopic enucleation is easy to perform in suitable lesions, and its advantage with respect to liver resection is the preservation of healthy parenchyma and liver ligaments. No outflow occlusion is needed, and the intraoperative bleeding is easily controlled. Technical aspects of laparoscopic enucleation are described and a review of the literature is also provided. PMID- 16340571 TI - Surgical clips: a nidus for foreign body reaction after hepatic resection. AB - This case report describes a patient who underwent segment IV hepatic resection and 7 months later developed an abdominal wall abscess. This was a foreign body reaction to the surgical clips. The patient required an exploratory laparotomy with debridement and excision of the inflammatory mass in the anterior abdominal wall. Although occurrence is rare, foreign body reactions to surgical clips have been reported, especially as a complication of laparoscopic surgery. PMID- 16340572 TI - Incidence and management of perineal hernia after laparoscopic proctectomy. AB - Perineal hernia (PH) is formed by the protrusion of intra-abdominal viscera through a defect in the pelvic floor. This is a rare complication after conventional abdominoperineal resection, pelvic exanteration, proctectomy, and other pelvic procedures. The purpose of the present paper is to report 4 cases of PH after laparoscopic abdominoperineal resection for rectal cancer and to review literature data about the incidence, predisposing factors, and treatment of this challenging problem. When added to other 3 cases previously reported in the Brazilian series of laparoscopic surgery, this group of 7 cases comprises a PH incidence of 3.5% after rectal resection procedures. Surgical treatment is indicated only in symptomatic patients with no signs of cancer recurrence. Proposed methods of surgical repair include abdominal, perineal, or combined approaches to the hernia in association with the use of autologous tissues or prosthetic meshes. Preventive measures are represented by closure of the pelvic peritoneum whenever possible, primary perineal suture and wound care to avoid infection. PMID- 16340573 TI - Laparoscopic resection of a torsioned appendix epiploica in a previously appendectomized patient. AB - Torsion of the appendices epiploicae is a rare condition that may present with acute abdominal pain and mimics appendicitis. We report a 20-year-old previously appendectomized man presenting with right lower abdominal quadrant pain. Abdominal ultrasonography showed a localized omental thickening in the right paracolic region. Contrast-enhanced computed tomography revealed well circumscribed fatty tissue adjacent to the cecum with heterogeneous hyperdense infiltration of the mesentery near the sigmoid colon. Diagnostic laparoscopy revealed 2-cm diameter torsioned and edematous fatty tissue floating on the omentum in the right lower quadrant. The torsioned mass was elevated, and a thick stalk was seen to be connecting the fatty tissue to the sigmoid colon. At this point, the torsioned fatty tissue was considered as a sigmoidal appendix epiploica that was elongated and neighboring on the previously operated-on region. The lesion was removed by laparoscopic means using 3 ports. Grossly, fat necrosis and internal bleeding were seen. Histopathologic analysis of the resected tissue demonstrated adipose tissue surrounded by fibrotic inflammatory changes with marked infiltration of numerous lymphocytes and histiocytes. In conclusion, torsion of appendices epiploicae should be included in the differential diagnosis of acute abdomen when evaluating patients with right lower quadrant pain and a history of appendectomy. Laparoscopic surgery provides definite diagnosis and prevents unnecessary open procedures for such lesions leading to peritoneal irritation. PMID- 16340574 TI - Laparoscopy-assisted surgical management of obscure gastrointestinal bleeding secondary to Meckel's diverticulum in a pediatric patient: case report and review of literature. AB - Despite advances in endoscopy and imaging, acute gastrointestinal (GI) bleeding of obscure origin in children presents a challenge to pediatric gastroenterologist. Bleeding Meckel's diverticulum (MD) commonly presents with acute episode of lower GI bleeding. A conventional diagnostic algorithm includes endoscopy, technetium 99m pertechnetate scintigraphy, angiography, and exploratory laparotomy. The advent of minimal access surgery prompts the use of laparoscopy for children with obscure GI bleeding. Laparoscopy assists in the diagnosis and can offer definitive treatment of patients with MD. Herein, we report a case of pediatric GI bleeding of obscure origin associated with MD that was successfully diagnosed and managed via laparoscopy. This article updates the current management for pediatric patients with obscure GI bleeding and the role of laparoscopy in the management of MD. PMID- 16340575 TI - Laparoscopic management of cecal injury from a misplaced percutaneous suprapubic cystostomy. AB - Iatrogenic bowel injury is a recognized complication of percutaneous suprapubic cystostomy. In the present report, we describe a case of misplacement of suprapubic catheter into cecum, in which laparoscopic technique was used successfully to identify and treat this condition. A 72-year-old woman with severe multiple sclerosis underwent her fourth change of suprapubic catheter 3 months after initial insertion. At the time of catheter change, the urologist performed a cystoscopy via the suprapubic tract and found feculent material in the presumed bladder. A diagnosis of colovesical fistula was made, and patient was referred to the acute surgical service. Cystogram via suprapubic catheter showed passage of contrast straight into colon in the region of cecum with no evidence of a fistula. Computed tomography of abdomen and pelvis confirmed the position of the catheter in the cecum. At laparoscopy, the cecum was seen to move when tension was applied to the suprapubic catheter, confirming the catheter tip and balloon in the lower pole of cecum. Laparoscopic transection of the cecum above the point of entry of the catheter was performed using an Endo GIA linear stapler. Under laparoscopic visualization, a new suprapubic catheter was inserted into the bladder. Intraoperative bleeding was minimal and the postoperative course was uneventful. PMID- 16340576 TI - Laparoscopic drainage of bilateral tuberculous psoas abscesses. AB - Psoas abscess secondary to Mycobacterium tuberculosis infection is rare in industrialized countries. Standard treatment options for psoas abscess of any etiology include percutaneous drainage under radiographic guidance and surgery, which is reserved for failure of conservative therapy. A case of bilateral tuberculous psoas abscesses is reported and a surgical method of drainage utilizing a totally extraperitoneal laparoscopic approach is described. PMID- 16340578 TI - A review of common practice in Dupuytren surgery. AB - Aspects of decision making, postoperative management, and complications routinely discussed with patients were investigated using a questionnaire completed by 141 hand surgeons of various grades who regularly manage Dupuytren disease. This account presents the results of this questionnaire study, and the literature is discussed in context. A brief description of operative technique in fasciectomy and dermofasciectomy is included. Thresholds for surgery did not differ greatly among the surgeons who completed the questionnaire. The training background and the number of operations performed by surgeons also did not generally influence surgical decision making. Postoperative care regimens were very similar. However, there was great variation in the complications routinely included in discussions with patients undergoing surgery for Dupuytren contracture. Recurrence and nerve damage were the only 2 mentioned by almost all surgeons. There was also diversity in what rates were quoted for complications, particularly recurrence (median 33%, range 0%-100%) and stiffness (median 10%, range 0%-100%). PMID- 16340579 TI - Simultaneous radial closing wedge and ulnar shortening osteotomy for distal radius malunion. AB - Closing wedge osteotomies are an attractive treatment option for distal radius malunion in patients with osteopenia. It does not require a separate-site bone graft, result-ing in decreased morbidity. However, this procedure has always been performed in combination with ulnar head resection to accommodate closure of corrective osteotomy and to address the issue of ulnocarpal abutment. Because ulnar shortening osteotomy is more physiological solution, it seemed advantageous to us to combine radial closing wedge osteotomy and ulnar shortening osteotomy. In this article, we present in detail our technique for treating patients with distal radius malunion. The patients are potentially osteopenic such as women aged over 45 years and are active at home, work, and/or recreation but are not involved in heavy physical work. Through volar approach an appropriate bone wedge is removed from the distal radius. A small volar T-plate is used to secure the osteotomized bone fragments. The aim of the radial osteotomy is to correct the dorsal angulation in the sagittal plane within a normal range of 1 degree to 2 degrees with reference to that of opposite wrist. Ulnar shortening osteotomy is performed by using transverse osteotomy and compression plating technique with an AO compression device. The ulnar variance is adjusted to neutral. Although the technique presented requires the surgeons to use careful plate and screw technique, early results have been encouraging, and patients with osteopenia can be treated successfully. Decreased grip strength which may be provoked by shorting of the forearm is acceptable. PMID- 16340580 TI - Distal radius fracture fixation with an intramedullary nail. AB - The purpose of this article is to describe a novel technique and implant system for fixation of unstable (Arbeitsgemeinschat fur Osteosynthesesfragen Types A and B) distal radius fractures. Currently the most common complications directly related to distal radius plate fixation includes tenosynovitis, tendon attrition, and rupture, often necessitating hardware removal. With the advent of this new device, a decrease in soft tissue complication is expected. The implant utilizes the principles of load sharing, subchondral screw divergence, and locked fixed angle fixation. It is inserted through a small skin incision at the radial styloid and does not further devascularize the fracture fragments. The limited surgical dissection and rigid fracture fixation allow for minimal postoperative immobilization and an early return of function. The authors believe that this system is a valuable addition to the arsenal of distal radius fracture treatment options and can quickly get patients back on the road of recovery. PMID- 16340581 TI - Cannulated variable pitch compression screw fixation of scaphoid fractures using a limited dorsal approach. AB - Scaphoid fractures are common in the young, active patient. Treatment is challenging because of the complex three-dimensional anatomy of the scaphoid and the tenuous blood supply. Traditionally, cast immobilization has been used for the management of non-displaced fractures with satisfactory outcomes reported in the literature. However, non-surgical treatment may result in a delayed union or nonunion particularly if the fracture is unstable, displaced, or involves the proximal pole. Recently there has been increased interest in the fixation of non displaced scaphoid fractures. The proposed advantages for operative treatment include avoiding the morbidity and inconvenience of prolonged cast immobilization and a lower incidence of delayed union or nonunion. A variety of surgical approaches for fixation of an acute scaphoid fracture have been described. The most common techniques include percutaneous fixation, arthroscopically assisted reduction and fixation, or open reduction and internal fixation via a volar approach. The senior author favors a limited dorsal approach with compression screw fixation of all proximal pole fractures as well as displaced and non displaced fractures of the waist region. The technique is simple permitting accurate screw placement in the central axis of the scaphoid, which is biomechanically advantageous and important for achieving union. PMID- 16340582 TI - A cosmetic technique for performing Basal joint arthroplasty. AB - Longitudinal scars perpendicular to the abductor skin creases of the wrist that result from basal joint arthroplasties can produce scarring that is distracting to both the patient and others. The author presents his technique for performing basal joint arthroplasty through a single transverse incision over the trapezium that blends with the normal creases of the wrist, producing a cosmetically acceptable scar. PMID- 16340583 TI - Optimization of flexor tenolysis using a suture. AB - In zone 2, complete release of all the peritendinous adhesions is almost impossible because of the complexity of the two tendons' three-dimensional arrangement under the pulleys. Resection of one slip of the flexor digitorum superficialis (FDS) changes this complex arrangement into a two parallel tendons pattern, which makes it much easier, particularly when using the suture technique. PMID- 16340587 TI - Psychometric properties of the mood and anxiety symptom questionnaire in patients with chronic pain. AB - OBJECTIVE: Previous studies have demonstrated that the Mood and Anxiety Symptom Questionnaire is able to discriminate between depressive and anxious symptoms. Factor analysis of the scale has revealed 3 factors: general distress, positive affect, and physiological hyper-arousal. However, the properties of the Mood and Anxiety Symptom Questionnaire have not been extensively examined in medical populations, in which the measurement of depressive and anxious symptoms is complicated by the presence of comorbid physical symptoms such as pain. Therefore, the purpose of the present study was to evaluate the psychometric properties of the Mood and Anxiety Symptom Questionnaire in patients with chronic pain. MATERIALS AND METHODS: A total of 230 persons with chronic musculoskeletal pain were recruited from a multidisciplinary pain clinic (n=110) and a large urban community (n=120). Participants completed the Mood and Anxiety Symptom Questionnaire and were interviewed for the presence of depressive disorders using the Structured Clinical Interview for the DSM-IV. Factor analyses, scale reliability analyses, and discriminative function analyses were performed on the data. RESULTS: Utilizing principal factor analysis and orthogonal rotation, the factor structure of the Mood and Anxiety Symptom Questionnaire in both chronic pain samples was similar to previously reported factor analyses. Three factors emerged: positive affect, general distress, and physiological hyper-arousal. Coefficients of congruence calculated from the factor structures of each subsample of persons with chronic pain suggested that the factors were highly similar (0.83-0.93). Given that the factor structures were comparable in both populations, a final analysis was conducted on the entire sample. Again, this analysis revealed a 3-factor solution. Examination of the factor loadings and item-to-total correlations revealed that some of the original scale items failed to load on or correlate with a specific factor, or correlated highly with 2 or more factors (thus lacking discriminatory power). A revised scale, eliminating items with low factors loadings and/or high cross loadings, was examined. Reliabilities for the subscales were high, and the subscales demonstrated the ability to predict clinical diagnoses of depression in a hypothesized fashion. DISCUSSION: The results support the utility of the Mood and Anxiety Symptom Questionnaire with samples of patients with chronic pain and provide further support for the tripartite model of anxiety and depression. PMID- 16340588 TI - Personality and pain-related beliefs/coping strategies: a prospective study. AB - OBJECTIVES: The ways in which people adjust to chronic pain has been partly attributed to pre-existing personality traits, but most evidence to date is from cross-sectional studies and mainly with arthritis groups. The present study examined the effects of 5 personality dimensions on measures of pain-related beliefs and catastrophizing assessed 9 months apart with a heterogeneous chronic pain sample. METHOD: One hundred forty-five patients with chronic pain presenting to a hospital pain center completed the NEO-Personality Inventory-Revised, 3 measures of pain related beliefs, the catastrophizing scale of the Coping Strategy Questionnaire, and a 0 to 10 pain scale. Nine months from the initial assessment, patients completed the same measures, apart from the NEO-Personality Inventory-Revised. RESULTS: Of the 5 personality dimensions studied, only neuroticism was related to the pain-related variables. Multiple hierarchical regression analyses revealed that neuroticism was a significant predictor of residualized change in pain self-efficacy beliefs and pain control appraisals over the time of the study, after controlling for initial values of both constructs. However, the effects were small, suggesting that other factors play a role in the determination of such beliefs, in addition to neuroticism. In contrast, neuroticism was not a significant predictor of residualized change in catastrophizing responses over the same period. The findings provide partial support for the hypothesis that pre-existing personality traits place some patients at risk for poor adjustment to chronic pain. PMID- 16340589 TI - Men with pelvic pain: perceived helpfulness of medical and self-management strategies. AB - OBJECTIVE: To assess the frequency of use of different treatments and pain management strategies and their perceived helpfulness in male patients with pelvic pain. METHODS: Approximately 1 month after a health maintenance organization visit for pelvic pain, 286 men (mean age 46.7 years) completed telephone interviews about their symptoms and treatments and pain management strategies used in the past year. Participants rated the helpfulness of each treatment and strategy used on 0 to 10 scales. RESULTS: Even though men with identified bacterial etiology were excluded from the study, antibiotic medication was the most commonly reported treatment (67% of patients) and rated as the second most helpful treatment [mean (SD)=6.3 (3.6)]. Opiates were rated as the most helpful treatment on average [mean (SD)=7.9 (2.1)], but were used by only 12% of patients. Substantial minorities of patients reported several behaviors as helpful, including urinating (reported as helpful by 26%), taking warm baths (23%), and drinking water (23%), although patterns of effects differed for men with versus without urinary symptoms. Activities most commonly reported as worsening symptoms were sitting (42%), walking/jogging (27%), and sexual activity (25%). DISCUSSION: Patients with male pelvic pain syndrome are commonly prescribed antibiotics, which they perceive as moderately helpful, despite the lack of scientific evidence of efficacy. Clinicians may find it useful to support patient use of safe, inexpensive, self-management approaches, especially warm baths, increased water intake, and avoidance of prolonged sitting. PMID- 16340590 TI - Clinically significant change in pain intensity ratings in persons with spinal cord injury or amputation. AB - OBJECTIVE: To examine the clinical significance of changes in chronic pain in 2 disability groups: spinal cord injury and lower limb amputation. METHODS: A reanalysis of 2 controlled clinical trials for pain in persons with disabilities. Eighty-two persons with spinal cord injuries and 34 persons with amputations provided pretreatment and posttreatment ratings of pain intensity on 0 to 10 numerical rating scales. After treatment, participants also rated the meaningfulness of any changes in pain they experienced. RESULTS: An average decrease of 1.80 points on the 0 to 10 numerical rating scales and percent decreases of about 36% corresponded to reports of a meaningful change in pain. There was no significant difference between samples, and also no differences due to gender or treatment condition. Age and pretreatment pain, however, were associated with the amount of pain decrease rated as meaningful. The change in pain intensity rated as meaningful was greater for older participants and participants with higher levels of pretreatment pain. Consistent with previous research, percent change scores were less biased by pretreatment pain than were absolute change scores. DISCUSSION: These findings are generally consistent with similar findings in other pain populations, and, in light of previous research, support an approximate 33% decrease in pain as a reasonable standard for meaningful change across chronic pain conditions. Percent change scores may be more useful for comparing the effects of pain treatments across samples or conditions because they are less biased by pretreatment pain level. PMID- 16340591 TI - Topically administered ketamine reduces capsaicin-evoked mechanical hyperalgesia. AB - BACKGROUND: The n-methyl-d-aspartate receptor antagonists such as ketamine relieve chronic pain but their oral and parenteral use is limited by the adverse effects. Experimental studies indicate that the peripheral n-methyl-d-aspartate receptors are involved in nociception. Recent clinical findings suggest that ketamine gel alleviates neuropathic pain, but no placebo-controlled randomized studies are available on the neurosensory effects of ketamine gel in experimental neurogenic pain. OBJECTIVES: The aim of this study was to assess the effects of topically applied ketamine using the intradermal capsaicin model in healthy volunteers. METHODS: Nine healthy subjects received ketamine and placebo gel on 3 occasions in a randomized, double-blind, and crossover manner. The concentration of ketamine was 50 mg/mL. One milliliter of gel was rubbed into the skin of both forearms 10 minutes before the intradermal injection of capsaicin (250 microg). Thereafter, the intensity and unpleasantness of spontaneous and evoked pain and dysesthesia was assessed up to 60 minutes using a 10-cm visual analog scale. Pain and dysesthesia were evoked using cotton gauze, a von Frey microfilament, and 38 degrees C, 42 degrees C, and 47 degrees C heat. Side effects were recorded, and individuals' subjective experiences were assessed with a standard questionnaire. RESULTS: Ketamine gel had no effect on immediate burning pain followed by the capsaicin injection. Both the intensity and unpleasantness of mechanical hyperalgesia was statistically significantly reduced by ketamine gel applied both on the left and right side. Neither tactile allodynia evoked by a brush nor thermal hyperalgesia were observed in any volunteer. No local or systemic side effects were observed. No patient reported any drug effects. DISCUSSION: A significant reduction of mechanical hyperalgesia was produced by topically and pre-emptively applied ketamine in healthy patients. We propose that the mechanism of action would be the reduction of central sensitization caused by the absorption of ketamine in circulation. PMID- 16340592 TI - Hypoalgesia in the referred pain areas after bilateral injections of hypertonic saline into the trapezius muscles of men and women: a potential experimental model of gender-specific differences. AB - OBJECTIVE: The study was to assess the local pain intensity, referred pain patterns, and changes in the pressure pain thresholds of the local and referred pain areas following bilateral injections of hypertonic saline into trapezius muscles and to delineate gender differences in pain profile and the possible modulation of pressure pain thresholds. METHODS: Fifteen healthy men and women each participated. Pain intensity and areas were assessed after bilateral injections. The pressure pain thresholds in the trapezius and posterolateral neck muscles (referred pain areas) were measured before and 7.5 and 15 minutes postinjection. RESULTS: The time to maximal pain intensity was significantly shorter after the second injection than after the first injection. Patients exhibited a bilaterally symmetrical distribution of pain areas that included the neck to shoulder angle and the posterolateral neck. Bilateral injections significantly elevated pressure pain thresholds in the right and left posterolateral neck muscles 7.5 and 15 minutes postinjection. In men but not women, the pressure pain thresholds measured in the posterolateral neck muscles 7.5 and 15 minutes after the second injection were higher than those after the first injection and also higher than those after the second injection in women, men tolerated more pressure than women at all time points. DISCUSSION: These findings indicate that both excitatory and inhibitory mechanisms modulate pain response characteristics in bilateral neck-shoulder pain conditions and suggest that there may be a more potent inhibitory control mechanism in men than women. This experimental model may be potentially used to detect gender differences in descending inhibition. PMID- 16340593 TI - Fear-avoidance beliefs, disability, and participation in workers and non-workers with acute low back pain. AB - OBJECTIVES: Fear-avoidance beliefs have been shown to be associated with disability and work status in patients with low back pain. Especially in acute low back pain, much research is needed to increase the knowledge concerning the role of fear-avoidance beliefs and its associated avoidance behavior in the influence on disability and the participation in daily and social life activities. The Fear-Avoidance Beliefs Questionnaire is developed to measure such beliefs and has become increasingly popular for use in primary care practice. The aim of the current study was: 1) to investigate the factor structure of the Fear Avoidance Beliefs Questionnaire in a population of patients with acute low back pain by means of confirmatory factor analysis; 2) to examine the relationship between fear-avoidance beliefs and perceived disability, as well as participation in daily and social life; and 3) to investigate whether perceived disability mediates the association between pain and participation and between fear avoidance beliefs and participation. METHODS: The study has a cross-sectional design and was performed on a sample of 615 patients with acute low back pain in primary care settings in The Netherlands. Patients completed a visual analogue scale for pain, the Roland Disability Questionnaire, the Fear-Avoidance Beliefs Questionnaire, a sociodemographic questionnaire, and 5 participation items. RESULTS: Confirmatory factor analysis corroborated the 2-factor structure of the Fear-Avoidance Beliefs Questionnaire as found by Waddell et al. Investigation of the association between fear-avoidance beliefs and perceived disability or participation was performed for workers and non-workers separately. The 2 subscales of the Fear-Avoidance Beliefs Questionnaire were significantly associated with perceived disability and participation, although there were differences between the samples. Perceived disability was significantly predicted by pain intensity and fear-avoidance beliefs. Perceived disability significantly predicted participation, and finally, the mediational role of perceived disability was supported. DISCUSSION: In the early stage of low back pain, the reduction of pain and fear-avoidance beliefs might increase the level of activity, which might foster increased participation in daily and social life activities. PMID- 16340594 TI - Electronic momentary assessment in chronic pain I: psychological pain responses as predictors of pain intensity. AB - OBJECTIVES AND METHODS: Electronic momentary assessment was employed to substantiate the relevance of psychological functioning in chronic pain. More than 7,100 electronic diaries from 80 patients with varying IASP classified types of chronic pain served to investigate to what extent fear-avoidance, cognitive and spousal solicitous and punishing pain responses explained fluctuations in pain intensity and whether patients with pre-chronic, recently chronic and persistently chronic pain differed in this regard. RESULTS: Psychological pain responses explained 40% of the total variance in pain intensity: almost 24% concerned pain variance that occurred between the CPD patients and 16% pertained to pain variance due to momentary differences within these patients. Separately tested fear-avoidance and cognitive responses each explained about 28% of the total pain variance, while spousal responses explained 9%. Catastrophizing emerged as the strongest pain predictor, followed by pain-related fear and bodily vigilance. Results did not differ with the duration of chronicity. DISCUSSION: Exaggerated negative interpretations of pain, and fear that movement will induce or increase pain strongly predicted CPD pain intensity. Spousal responses assessed only when the spouse was with the patient who at that moment was in actual pain-may more strongly affect immobility due to pain than pain intensity per se (see part II of the study). The findings substantiate the importance of catastrophizing, fear and vigilance identified primarily in low back pain and extend this to other forms of chronic pain. The compelling evidence of momentary within-patients differences underscores that these must be accounted for in chronic pain research and practice. PMID- 16340595 TI - Electronic momentary assessment in chronic pain II: pain and psychological pain responses as predictors of pain disability. AB - OBJECTIVES AND METHODS: More than 7,100 electronic diaries from 80 patients with chronic pain (mean: 89.3, range 30-115) entered multilevel analyses to establish the statistical prediction of disability by pain intensity and by psychological functioning (fear avoidance, cognitive, and spousal pain responses). We also tested the differences between pre-chronic, recently chronic, and persistently chronic pain in the prediction of disability (impaired physical and mental capacity, pain interference with activities, immobility due to pain). RESULTS: Pain intensity explained 8% to 19% of the disability variance. Beyond this psychological functioning explained 7% to 16%: particularly fear-avoidance and cognitive pain responses predicted chronic pain disorder disability; spousal responses predicted immobility better than other aspects of disability. Immobility due to actual pain occurred infrequently. When it did, however, it was better predicted by avoidance behavior in the patient and by spousal discouragement of movement than by actual pain intensity. The prediction of immobility due to pain by, respectively, avoidance behavior and catastrophizing was better in chronic pain (>6 months) and that of physical impairment by catastrophizing better in persistently chronic pain (>12 months) than in pain of shorter duration. DISCUSSION: The psychological prediction of chronic pain disorder disability was determined beyond that accounted for by pain intensity. Nonetheless, psychological functioning explained substantial variance in chronic pain disorder disability. The psychological prediction of immobility and physical impairment was stronger with longer pain duration. Patient characteristics and momentary states of disability-and in particular of immobility-should be carefully distinguished and accounted for in chronic pain disorder. PMID- 16340596 TI - Tunneled epidural catheter infections in noncancer pain: increased risk in patients with neuropathic pain/complex regional pain syndrome. AB - BACKGROUND AND OBJECTIVE: Tunneled epidural catheters are often used to control pain and facilitate rehabilitation in patients with regional pain syndromes. A cohort retrospective study design was undertaken to evaluate the risk of catheter related infection in patients who underwent a TEC placement to manage their chronic noncancer regional pain over a 5-year period. METHODS: There were 260 tunneled epidural catheters placed in 218 patients accounting for a total of 10,985 catheter-days. Of these, 230 catheters were placed in patients with neuropathic pain (90% of whom had complex regional pain syndrome) and 30 catheters in patients with somatic pain. RESULTS: There were 24 epidural space infections in symptomatic patients, 23 of whom were in the neuropathic pain group 22 had complex regional pain syndrome. Additionally, tunneled epidural catheters had to be discontinued in 34 patients because of superficial infection or suspicion of infection; 33 of these were in the neuropathic pain group. The differences in the infection rates were significantly higher in the neuropathic pain group compared to the somatic group. When analyzed to an infection index per 1,000 catheter-days, the rate of infection rate was 5.51 for the patients with neuropathic pain and 2.43 for the patients with somatic pain. The rates for deep and superficial infections were 2.26 and 3.25, respectively, per 1000 catheter days for the neuropathic pain group compared to 1.22 for both deep and superficial infections in the patients with somatic pain. There were 6 frank epidural abscesses upon contrast-enhanced magnetic resonance imaging examinations of the spine, 1 epidural phlegmon and 2 patients displayed mild or questionable epidural enhancement on magnetic resonance imaging, suggestive of epidural inflammation. All these magnetic resonance imaging abnormalities were detected in patients with complex regional pain syndrome. Two of the patients with epidural abscesses underwent surgical exploration and drainage of the epidural abscess, though no neurologic deficits were observed in any of the patients. CONCLUSIONS: The higher risk of tunneled epidural catheter infection observed in patients with neuropathic pain (particularly complex regional pain syndrome) warrants further study. PMID- 16340597 TI - The effect of small doses of botulinum toxin a on neck-shoulder myofascial pain syndrome: a double-blind, randomized, and controlled crossover trial. AB - OBJECTIVES: Myofascial pain syndrome is a common cause of muscular pain in the shoulder-neck region. Injections of large amounts of botulinum toxin A have been found to be beneficial for the alleviation of myofascial pain, but large doses of this toxin may cause paresis of the muscle and other adverse events. The aim of this work was to determine the effect of small doses (5 U) of botulinum toxin A (BTA) injected directly into the painful trigger points of the muscles, using a double-blind crossover technique. METHODS: On the basis of the empirical criteria proposed for diagnosis of myofascial pain syndrome, 31 patients suffering from myofascial pain in the neck-shoulder region were studied. The patients received either botulinum toxin A or physiological saline injections on 2 occasions 4 weeks apart. The total dose varied from 15 to 35 U of botulinum toxin A [28+/- 6 U (mean+/- SD)]. The follow-up measurements were carried out at 4 weeks after each treatment. Neck pain and result of treatment were assessed with questionnaires. The pressure pain threshold was determined using a dolorimeter. RESULTS: Neck pain values decreased from 4.3+/- 2.4 to 3.3+/- 2.0 after saline injections and from 4.1+/- 2.1 to 3.3+/- 2.2 after botulinum toxin A. The pressure pain threshold values increased from 5.2+/-1.6 to 5.9+/-1.5 and from 5.7+/-1.6 to 5.9+/-1.6 after injections with saline and botulinum toxin A, respectively. No statistically significant changes in the neck pain and pressure pain threshold values occurred between the botulinum toxin A and saline groups. After the first injections, the subjective result of treatment was significantly (P=0.008) in favor of botulinum toxin A, and after the second injections, the subjective result was better for saline, but the difference was not statistically significant (P=0.098). There was no significant difference in the prevalence of side effects between saline and botulinum toxin A. CONCLUSIONS: Our study shows that there was no difference between the effect of small doses of botulinum toxin A and those of physiological saline in the treatment of myofascial pain syndrome. PMID- 16340598 TI - The validity of the neuropathic pain scale for assessing diabetic neuropathic pain in a clinical trial. AB - OBJECTIVES: In controlled trials of analgesics for the treatment of neuropathic pain, the primary outcome variable is most often a measure of global pain intensity. However, because neuropathic pain is associated with a variety of pain sensations, the effects of analgesic treatments on different sensations could go undetected if specific pain qualities are not assessed. This study sought to evaluate the utility of assessing the multiple components of neuropathic pain in an analgesic clinical trial. METHODS: One hundred fifty-nine subjects with diabetes-related foot pain were randomly assigned to receive an active analgesic (controlled-release oxycodone) or matching placebo for 6 weeks. A multidimensional measure of neuropathic pain, the Neuropathic Pain Scale (NPS), was administered before, during, and after study treatment. RESULTS: Relative to placebo, the opioid analgesic produced statistically significantly greater decreases in global pain intensity, pain unpleasantness, and sharp, dull, and deep pain sensations. Responder analyses indicated a higher rate of responding to the opioid condition, relative to placebo, for intense, unpleasant, deep, and surface pain. The opioid analgesic did not significantly reduce hot, cold, itchy, or sensitive pain sensations compared with placebo in either analysis. CONCLUSIONS: These findings support the utility of the NPS for characterizing the multidimensional nature of the neuropathic pain experience and for detecting changes in neuropathic pain with treatment. PMID- 16340599 TI - Thermal and tactile sensory deficits and allodynia in a nerve-injured patient: a multimodal psychophysical and functional magnetic resonance imaging study. AB - OBJECTIVE: A case study was conducted to examine a patient with chronic neuropathic pain of the right foot following peripheral nerve injury and characterize associated sensory abnormalities. METHODS: Multimodal psychophysical examination of the patient's affected and nonaffected foot included thermal sensibility, dynamic touch, and directional sensibility. In addition, we used functional magnetic resonance imaging to study cortical representation of brush evoked allodynia. RESULTS: Detailed psychophysical examination revealed substantial deficits in warm, cool, and tactile perception on the injured foot. These findings indicated severe dysfunction of perceptual processes mediated by A beta, A delta, and C fibers. Despite reduced tactile perception, light touch evoked a deep burning pain in the foot. Functional magnetic resonance imaging during brushing of the patient's injured foot showed that tactile allodynia led to activation of several cortical regions including secondary somatosensory cortex, anterior and posterior insular cortex, and anterior cingulate cortex. Brushing of the patient's nonaffected foot led to fewer activated regions. DISCUSSION: The profound sensory disturbances suggest a possible deafferentation type of tactile allodynia mediated by changes within the central nervous system, such as a disruption of normal tactile or thermal inhibition of nociception. The functional magnetic resonance imaging data suggest that tactile allodynia is represented in similar brain regions as experimental pain. PMID- 16340600 TI - Basal blood flow in complex regional pain syndrome does not necessarily indicate vasoconstrictor nerve activity. AB - OBJECTIVES: To quantitatively investigate simultaneous skin blood flow and sweating in a patient with complex regional pain syndrome. METHOD: Using one probe, skin blood flow and sweating were measured on the affected and non affected sides at baseline and after activation of sympathetic system in a 47 year-old woman with complex regional pain syndrome type I of the left hand. RESULTS: Basal sweating, sympathetic sweat response, basal blood flow, sympathetic flow response, and attrition rate of blood flow (the ratio of sympathetic flow response to basal blood flow) of the affected side were greater than those on the non-affected side. CONCLUSIONS: Although sympathetic nerve activity (sympathetic flow response) appeared greater on one side, basal blood flow was also greater on that side. These contradictory results suggest that some factor or factors increase basal blood flow despite a higher sympathetic nerve tone on the side. Comparing basal blood flow on one side to the contralateral side does not necessarily indicate lesser vasoconstrictor nerve activity on that side. Advantages of simultaneous measurement of skin blood flow and sweating with a single probe may make the method useful. PMID- 16340601 TI - Plugging into Bush's Energy Policy. PMID- 16340602 TI - Validation of ICNIRP estimates of toxicity thresholds for NIR (785 nm) light in the retinas of pigmented rabbits. AB - The International Commission on Non-Ionizing Radiation Protection (ICNIRP) established eye-safe exposure limits for laser light based on estimates of the 50% exposure dose (ED50) for retinal damage in animals. The ED50 has not been determined for continuous-wave near-infrared (NIR) light, as may be used in instruments for spectroscopic analysis of the eye. The purpose of this study is to determine the ED50 of 785-nm light delivered by a Raman spectroscopy instrument on the retinas of pigmented rabbits, and to compare that ED50 to the ICNIRP estimates. Light of varying power was focused into the anterior chambers of rabbits for 4, 20, or 300 s. Estimates of spot size on the retina were determined by modifying the schematic rabbit eye with actual axial length and refractive error measurements. Masked examiners determined retinal burns from fundus photographs and fluorescein angiograms made immediately after exposure and again 48-72 h later. Thirty-five of 86 exposed eyes had retinal burns. In 12 of these the burn was not visible until 48-72 h. In 6 the burn was visible on color photographs, but not on fluorescein angiography. The ED50 retinal power density from probit analysis was 14,513 mW cm(-2), 10,369 mW cm(-2), and 5,237 mW cm(-2) for 4-s, 20-s, and 300-s exposures, respectively. Thus, the ED50 for retinal burns from 785-nm light is approximately 7 and 7.5 times the ICNIRP permitted limits for 4- and 20-s exposures, and 5.6 times the limit for 300-s exposures. This is consistent with the ICNIRP intention to set power limits several times less than ED50. PMID- 16340603 TI - Dietary iodine intake in residents of northwestern regions of ukraine contaminated by the chernobyl accident. AB - Daily iodine intake in Ukrainian subjects of northwestern regions was estimated in relation to the health effects on inhabitants after the Chernobyl accident. Total diets were collected from 106 locations for children and adult males by a duplicate portion study. Iodine was rapidly determined by inductively coupled plasma mass spectrometry after chemical separation. Iodine concentration on a dry basis for Ukrainians was 0.11 microg g(-1) and the daily iodine intake was in the range of 2.80-372 microg per person. The median, geometric mean, and standard deviation were 28.1, 32.7, and 2.51 microg, respectively. The yearly trend of the intake had almost no change. Regional differences would be expected to exist among the 10 areas of the Ukraine, but no clear differences appeared in the present findings. Daily iodine intake in Ukrainians was lower than the recommended dietary intake (RDI) allowance (150 microg), and its lack would be related to the high prevalence of goiter in the country. PMID- 16340604 TI - Radioactive materials in biosolids: dose modeling. AB - The Interagency Steering Committee on Radiation Standards (ISCORS) has recently completed a study of the occurrence within the United States of radioactive materials in sewage sludge and sewage incineration ash. One component of that effort was an examination of the possible transport of radioactivity from sludge into the local environment and the subsequent exposure of humans. A stochastic environmental pathway model was applied separately to seven hypothetical, generic sludge-release scenarios, leading to the creation of seven tables of Dose-to Source Ratios (DSR), which can be used in translating from specific activity in sludge into dose to an individual. These DSR values were then combined with the results of an ISCORS survey of sludge and ash at more than 300 publicly owned treatment works, to explore the potential for radiation exposure of sludge workers and members of the public. This paper provides a brief overview of the pathway modeling methodology employed in the exposure and dose assessments and discusses technical aspects of the results obtained. PMID- 16340605 TI - Long-term monitoring of radioactivity in surface air and deposition in New York State. AB - Gross-beta activities have been determined weekly for 22 y from filtered atmospheric aerosols at seven sites in New York State. The activities, ranging from 0.1 to 0.9 mBq m(-3), varied seasonally and were evaluated in terms of meteorological factors. Cosmogenic beryllium (7Be) concentrations were determined quarterly on the air filters and weekly in deposition collected at one site. Over 98% of the air filters contained observable activity concentrations of 7Be (mean of 3 mBq m(-3)) and 210Pb (mean of 1 mBq m(-3)). However, only 20% of deposition samples contained Be concentrations above analytical detection limits. Tritium (3H) concentrations were below detection limits in deposition samples at the background site, but were present on most samples collected near a H-processing facility. Measurements of 131I were conducted weekly on charcoal canisters, with only one site showing observable concentrations (mean of 1 mBq m(-3)), due to nearby incineration of dried municipal sludge containing patient waste from hospital treatments. PMID- 16340606 TI - Thyroid dose measurements for staff involved in modified barium swallow exams. AB - Following a successful claim for damages by a speech pathologist, who was diagnosed with thyroid cancer after a period of employment including involvement in modified barium swallow procedures, a detailed study on occupational dose was commissioned. In this study radiation doses were measured routinely over 8 clinical periods using TLD100H dosimeters for staff involved with modified barium swallow radiological studies at three hospitals. The regions monitored included the unshielded thyroid, waist under apron, and axilla. Average unshielded thyroid doses for the speech pathologists were measured as 0.017, 0.003, and nil mGy per procedure over the hospitals surveyed, with dose measurements also found for the unshielded thyroid of the radiologist and nurse. This compares to thyroid doses of 0.200 and 0.055 mGy per procedure typically for cardiologists and orthopedic surgeons involved in radiological work, respectively. The use of a thyroid collar will substantially reduce this dose by a factor of about 40. While the worst-case doses are very small, attention should be given to education for staff involved in radiological procedures as well as ensuring that sound procedure protocols are in place and used. PMID- 16340607 TI - Prediction of tritium level in agricultural plants after short term exposure to HTO vapor and its comparison with experimental results. AB - This paper describes a dynamic compartment model for evaluating the tritium level in agricultural plants after a short-term exposure to HTO vapor and its comparison with experimental results to test the predictive accuracy of the model. The model uses a time-dependent growth equation of a plant so that it can predict the contamination level of tritium depending on the stage of the growth of the plant, which is a major difference from some other compartment models using a constant crop yield. The model is able to calculate the time variable concentrations of the compartments representing the atmosphere, soil, and plants of four categories including leafy vegetables, root vegetables, grains, and tuber plants. Experimental results include the tissue free water tritium (TFWT) and the organically bound tritium (OBT) concentration of rice, soybean, cabbage, and radish exposed to HTO vapor for 1 h in the daytime at different growth stages. The model predictions showed that the model could simulate well not only the time dependent tritium concentration of the plants but also the effect of the growth stage of the plant at the exposure time. Comparison of the model predictions with the experimental results suggested that the model could predict reasonably well the observed TFWT and OBT concentrations of the plants considered. PMID- 16340608 TI - Calculation of dose coefficients for radionuclides produced in a spallation neutron source utilizing NUBASE and the evaluated nuclear structure data file databases. AB - Based on a mercury spallation neutron source target, the UNLV Transmutation Research Program has identified 72 radionuclides with a half-life greater than or equal to a minute as lacking an appropriate reference for a published dose coefficient according to existing radiation safety dose coefficient databases. A method was developed to compare the nuclear data presented in the ENSDF and NUBASE databases for these 72 radionuclides. Due to conflicting or lacking nuclear data in one or more of the databases, internal and external dose coefficient values have been calculated for only 14 radionuclides, which are not currently presented in Federal Guidance Reports Nos. 11, 12, and 13 or Publications 68 and 72 of the International Commission on Radiological Protection. Internal dose coefficient values are reported for inhalation and ingestion of 1 microm and 5 microm AMAD particulates along with the f1 values and absorption types for the adult worker. Internal dose coefficient values are also reported for inhalation and ingestion of 1 microm AMAD particulates as well as the f1 values and absorption types for members of the public. Additionally, external dose coefficient values for air submersion, exposure to contaminated ground surface, and exposure to soil contaminated to an infinite depth are also presented. PMID- 16340609 TI - A 244Cm irradiator for protracted exposure of cultured Mammalian cells with alpha particles. AB - A 244Cm alpha-particle irradiator was designed and constructed for radiobiological studies where protracted exposure at a low dose rate of cultured mammalian cells is required. It allows irradiation of a cell monolayer attached to the Mylar bottom of a specially designed Petri dish of 56 mm diameter (approximately 25 cm(2) area). The irradiator is based on a 20-mm-diameter stainless steel chamber containing a 148 kBq 244Cm source. The chamber, flushed with helium gas at a pressure kept slightly above the external pressure, is inserted into a cell incubator where temperature and CO2 concentration are controlled. Spectrometric and dosimetric characterization of the irradiator was carried out by means of an ion-implanted-silicon charged-particle detector, CR39 detectors, and Monte Carlo simulations with the TRIM code. Average LET of particles incident on the cells at the center of the Petri dish was evaluated to be 120 keV microm(-1) at 59 mm from the source, and the average dose rate was 5.69 x 10 Gy s(-1), with +12% and -8% variations at the center and the edge, respectively. The irradiator has been successfully tested and used for several experiments involving 16-d exposure of human fibroblasts monolayers. PMID- 16340610 TI - Quality assurance methods and procedures used to verify consistency in calculating dose coefficients. AB - The development of a spallation neutron source with a mercury target will lead to the production of rare radionuclides. The dose coefficients for many of these radionuclides have not yet been published. A collaboration of universities and national labs has taken on the task of calculating dose coefficients for the rare radionuclides using the software package DCAL. The working group developed a procedure for calculating dose coefficients and a quality assurance (QA) program to verify the calculations completed. The first portion of this QA program was to verify that each participating group could independently reproduce the dose coefficients for a known set of radionuclides. The second effort was to divide the group of rare radionuclides among the independent participants in a manner that assured that each radionuclide would be redundantly and independently calculated, and the results subsequently be submitted for publication in a separate manuscript. The final aspect of this program was to resolve any discrepancies arising among the participants as a group. The output of the various software programs for six QA radionuclides, 144Nd, 201Au, 50V, 61Co, 41Ar, and 38S were compared among all members of the working group. Initially, a few differences in outputs were identified. This exercise identified weaknesses in the procedure, which has since been revised. After the revisions, dose coefficients were calculated and compared to published dose coefficients with good agreement. The present efforts involve generating dose coefficients for the rare radionuclides anticipated to be produced from the spallation neutron source should a mercury target be employed. PMID- 16340615 TI - Facilitators and barriers to continuing healthcare after jail: a community integrated program. AB - A cooperative, community-oriented "public health model of correctional healthcare" was developed to address the needs of persons temporarily displaced into jail from the community, and to improve the health and safety of the community. It emphasizes 5 key elements: early detection, effective treatment, education, prevention, and continuity of care. In the program, physicians and case managers are "dually based"-they work both at the jail and at community healthcare centers. This, together with discharge planning, promotes continuity of care for inmates with serious and chronic medical conditions. This report characterizes the health status and healthcare in this group, and identifies facilitators and barriers to engagement in primary medical and mental health care after release from jail. PMID- 16340616 TI - Changes needed in the healthcare system: perspectives of lay health workers. AB - Lay health workers can play a crucial role in connecting the community-in-need to the healthcare system. This article provides insights into how lay health workers, selected from underserved communities, view the issues faced in accessing the healthcare system. A sample of lay health workers responded to a questionnaire used to identify barriers they, themselves, experienced as well as get their perspectives on obstacles faced by their clients. The results demonstrated that lay health workers perceived themselves less affected by barriers as compared with their clients in regard to their provider relationship, getting health information, and their own personal attitudes and beliefs about healthcare. In addition, focus group discussions yielded recommendations and potential solutions to reduce barriers and improve the healthcare system, which included improved access to the facilities, management, scope of services offered, and provider behaviors. These results may benefit the efforts of healthcare professionals and researchers by enhancing their knowledge of, and facility to utilize and deploy, community resources, and, in turn, will assist underserved populations to better negotiate the system and obtain the services they need the most. PMID- 16340617 TI - Comparative effectiveness of health centers as regular source of care: application of sentinel ACSC events as performance measures. AB - A 4-state (Alabama, California, Georgia, Pennsylvania) retrospective analysis of claims data from 1.6 million Medicaid beneficiaries to assess the performance of community health centers compared with other Medicaid providers (office-based and hospital-based practices) served as a regular source of care to Medicaid beneficiaries, each with at least one diagnosed ambulatory care-sensitive condition (ACSC). The health centers compared with the other Medicaid providers experienced one third fewer sentinel ACS events: 5.7 and 8.2 ACS admissions and 26.1 and 37.7 ACS emergency visits, respectively, per 100 persons. Controlling for case mix and other factors, the logistic regression results for sentinel events indicated that Medicaid beneficiaries who relied on health centers for primary care were significantly less likely to experience an ACS admission (OR = 0.89, P < .0001) or an ACS emergency visit (OR = 0.81, P < .0001) than the Medicaid beneficiaries who relied on other Medicaid providers. Sentinel ACS events can serve as efficient measures for assessing provider performance and comparing effectiveness of regular sources for primary care. PMID- 16340618 TI - Translating research to action: improving physician access in public insurance. AB - Policymakers continue to struggle with how to assure adequate access to physician services in public programs like Medicaid, State Children's Health Insurance Program, or other public coverage programs. In this article, we synthesize available research on this topic and provide a framework that policymakers may find useful in identifying and measuring barriers to care access, determining where and why problems exist, and identifying how to intervene. Using our experience constructing the framework, we also consider what observations can be drawn from this experience for those interested in the challenge of moving the insights from research to practice. PMID- 16340619 TI - Mental disorders and medical care utilization of VA ambulatory care patients: the veterans health study. AB - In this article, the authors examine patterns of medical services use among the Department of Veterans Affairs (VA) ambulatory care patients who screened positive for posttraumatic stress, depression, or alcohol-related disorders. On the basis of research linking mental disorders with increased use of medical services, the authors hypothesize that even after controlling for age and medical disease comorbidity, patients in VA ambulatory care who screen positive for targeted mental disorders would be more likely to use VA medical services and have higher rates of such use than those who did not screen positive. Baseline data were obtained from the Veterans Health Study, a longitudinal investigation of veterans' health. Four Boston-area VA ambulatory care facilities were used as study sites. A random sample of 2425 participants (mean age = 62) was drawn from male VA ambulatory care patients screened for eligibility during specified periods. Screening measures were Center for Epidemiological Studies-Depression Scale for depression, Posttraumatic Stress Disorder Checklist for posttraumatic stress disorder, and CAGE Questionnaire for alcohol-related disorders with endorsement of prior year consumption for alcohol-related disorders. Prior medical services use was assessed by self-report. Although unadjusted analyses of medical services use revealed clear effects of the screening presence of mental disorders on most outcomes, after adjusting for age and medical comorbidity, almost all these effects were reduced, and some previously nonsignificant results became significant. Findings suggest that healthcare policy and risk adjustment predicated upon the presumed relationship between mental disorders in the aggregate and medical services use should reconsider the important contributions of age, comorbid medical disorders, and specific mental disorder diagnoses. PMID- 16340620 TI - The illness burden of alcohol-related disorders among VA patients: the veterans health study. AB - Little is known about the illness burden associated with alcohol-related disorders (ie, problem drinking, alcohol abuse, and alcohol dependence) among patients in outpatient medical care. The objective of this study was to examine several aspects of illness burden-medical comorbidities, patterns of health services use, and functional status-among Veterans Health Administration (VA) ambulatory care patients with alcohol-related disorders. Male participants (N = 2425) were recruited at 1 of 4 Boston-area VA outpatient clinics. They completed self-report screening measures of current alcohol-related disorders (CAGE score > or =2 with past year alcohol consumption), health behaviors, medical comorbidities, and functional status (SF-36). A medical history interview, which assessed comorbid conditions and use of recent health services, was also administered. Screening criteria for current alcohol-related disorders were satisfied by 12%; however, only 40% of these reported ever receiving treatment specifically for alcohol-related disorders. Patients who screened positive for alcohol-related disorders reported significantly greater limitations in mental health function, longer hospitalizations for medical care in the prior year, and fewer outpatient medical visits in the previous 3 months. Findings suggest considerable illness burden associated with alcohol-related disorders among VA ambulatory care patients. Efforts to increase detection and treatment of alcohol related disorders may lessen the illness burden and cost of alcohol-related disorders. PMID- 16340621 TI - Posttraumatic stress disorder and health status: the veterans health study. AB - This article examines the association between self-reported prevalence of posttraumatic stress disorder (PTSD) and health status in a sample of 2425 male Department of Veterans Affairs (VA) ambulatory care patients who participated in the Veterans Health Study. Participants were recruited at 1 of 4 VA outpatient clinics in the Boston area. They completed self-report measures of PTSD (using the PTSD Checklist and measures of exposure to traumatic events), depression (using the Center for Epidemiologic Studies--Depression scale), and health status (using the Short-Form-36) and a medical history interview assessing 22 conditions and a history of psychiatric treatment. The screening prevalence of PTSD was 20.2% among all patients (24.3% among those exposed to traumatic events); another 15.5% met the criteria for depression but not PTSD. The health status of patients with either PTSD or depression was significantly worse than that of patients with neither disorder, even after controlling for age, education, and number of comorbid medical conditions. Patients with PTSD reported more medical conditions than did other patients. Patients with PTSD currently in mental health treatment had worse health status than did those who reported no treatment; the health status of patients who reported past mental health treatment was generally comparable to that of those with no treatment. The prevalence and comorbidity of PTSD among this sample of VA ambulatory care patients were higher than previously reported among samples of community-residing adults. The association of PTSD with health status was substantial, suggesting that the burden of PTSD is at least comparable to, and may be worse than, that of depression. Mental health treatment alleviated some of this burden. The potential impact of PTSD on health status should be more widely recognized. PMID- 16340622 TI - Validation of the primary care alcohol severity measure. AB - Alcohol-related disorders are common in primary care settings; many primary care physicians are ill-equipped to manage patients with alcohol-related disorders. The objective of this prospective cohort study was to develop and validate a patient-based measure, the Primary Care Alcohol Severity Measure, to determine which primary care patients with alcohol-related disorders would benefit from referral to alcohol treatment services. Four Boston-area Department of Veterans Affairs ambulatory care clinics were chosen as study sites. Two hundred seventy eight male patients, mean age 55.5 years, 89.9% Caucasian, 42.5% married, all with CAGE Questionnaire scores greater than or equal to 2 and drinking within past year, participated in the study. We developed a multidimensional, 30-item measure that contained 2 subscales that assessed 2 symptom clusters of alcohol related disorders: Physical and Behavioral. Each subscale's score was higher (more severe) for patients with a current Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised diagnosis of alcohol dependence or abuse: P < .01 for the physical subscale and P < .0001 for the behavioral subscale. Patients with more physical symptoms had poorer physical and mental health status, whereas patients with more behavioral symptoms had poorer mental health status. Scores on the 2 subscales, along with age and history of prior treatment, predicted the use of alcohol treatment services in the following year: c = 0.90 in logistic regression. The Primary Care Alcohol Severity Measure is a valid measure of alcohol severity in primary care patients and predicts the use of alcohol treatment services. It is relatively brief and easy to use, requiring only standard medical history items and patient reports of behavioral symptoms. It may be a useful tool to improve the quality of care for primary care patients with alcohol-related disorders. PMID- 16340623 TI - Africa's health worker shortage undermines global health goals. PMID- 16340625 TI - To biopsy or not to biopsy: that is no longer the question! PMID- 16340626 TI - Celiac disease: current concepts in diagnosis and treatment. AB - Celiac disease is a complex autoimmune enteropathy that affects the small bowel in genetically predisposed individuals. It is thought that celiac disease is the result of an inappropriate T cell-mediated immune response against ingested gluten protein. The characteristic lesion of the small intestinal mucosa includes loss of absorptive villi and infiltration of the lamina propria with inflammatory cells. The clinical presentation of celiac disease varies greatly depending on patient's age, duration and extent of the disease, and the presence of extraintestinal manifestations. Unfortunately, most patients with celiac disease have either silent or atypical presentations, thus escaping diagnosis for several years. Medical nutrition therapy with lifelong adherence to a strict gluten-free diet is the only accepted treatment of celiac disease. Individuals at risk for this entity should undergo appropriate serologic testing, but there is no evidence to support mass screening. PMID- 16340627 TI - Helicobacter pylori: what do we still need to know? AB - Helicobacter pylori is a common gastric infection that causes serious complications in a minority of individuals. A great deal is known about the disease, but many unanswered questions remain. Among these, perhaps the most enigmatic is that we do not know how the organism is transmitted. We are uncertain why the prevalence of the disease is falling within the developed population. There is still debate as to its relationship with gastroesophageal reflux disease. It is unclear why there are differences in the international incidence of gastric cancer. Similar uncertainties relate to the reasons why the complications of H. pylori infection have changed over time. In this article, I have hypothesized that a number of these unanswered questions may be related to a putative increase in gastric acid secretion that may have taken place during the past 200 years. To date, there is little confirmatory evidence for this, and it remains a fascinating area that merits more scientific research. PMID- 16340628 TI - Day-to-day variability in acid reflux patterns using the BRAVO pH monitoring system. AB - BACKGROUND & GOALS: The wireless pH monitoring system such as the BRAVO pH system is a significant advancement in the evaluation of patients with gastroesophageal reflux because of its potentially better tolerability and the ability to record data over a 48-hour period. The aim of our study was to evaluate safety, performance, tolerability, and day-to-day variability in acid reflux patterns using the BRAVO pH system. METHODS: A total of 90 consecutive patients (48 men and 42 women) with persistent reflux symptoms underwent BRAVO pH capsule placement from October 2002 to August 2003 at a tertiary care hospital. The BRAVO pH capsule was deployed 6 cm proximal to the squamocolumnar junction under endoscopic guidance. The pH recordings over 48 hours were obtained after uploading data to a computer from the pager-like device that recorded pH signals from the BRAVO pH capsule. RESULTS: Successful pH data over 48 hours was obtained in 90% of patients. Nearly two thirds of patients experienced a variety of symptoms ranging from a foreign body sensation to chest discomfort or pain. Four patients had severe chest pain, 3 of whom required endoscopic removal of the BRAVO pH capsule. In 74.4% of patients, number of reflux events as well as time (%) pH<4 correlated from the first 24-hour period to the second 24-hour period. However, in 28% of patients, no predictable pattern of (%) time pH<4 in the supine position was reproduced from one 24-hour period to the next 24-hour period. CONCLUSIONS: The BRAVO pH system appears a safe and effective method of recording esophageal acid exposure. It is an acceptable alternative for patients who are unwilling or unable to tolerate nasopharyngeal catheter-based pH studies, and it has a potential advantage of the 2-day recording period. PMID- 16340629 TI - Factors associated with failure of initial endoscopic hemoclip hemostasis for upper gastrointestinal bleeding. AB - BACKGROUND: Endoscopic hemoclip is widely used for the management of bleeding peptic ulcers. The major difficulty in clinical application of the hemoclip is deployment to the lesion during initial hemostasis. The aim of this study was to define factors associated with the failure of endoscopic hemoclip for initial hemostasis of upper GI bleeding. PATIENTS AND METHODS: From January to December 2003, we prospectively studied 77 randomized patients with clinical evidence of upper GI bleeding due to either active bleeding or a visible vessel identified by upper GI endoscopy in our emergency department. RESULTS: Among the 77 patients, 13 (16.9%) failed treatment (Group 1) and 64 (83.1%) were successfully (Group 2) treated by endoscopic hemoclip for lesions related to upper GI bleeding. There were no differences due to gender, blood pressure, initial heart rate, and hemoglobulin before or after endoscopic treatment, platelet count, serum creatinine, and albumin between groups. The mean age of Group 1 was higher than that of Group 2 (73.31+/-9.38 years vs. 65.41+/-16.45 years, respectively; P=0.083). Most patients who did not achieve initial hemostasis by endoscopic hemoclip had upper GI lesions over the gastric antrum and duodenal bulb. Among the 13 patients who failed to achieve endoscopic hemoclip initial hemostasis, four lesions were located over the posterior wall of the antrum, and four lesions over the lesser curvature side of the duodenal bulb. CONCLUSION: Endoscopic hemoclip is an effective hemostatic method for upper GI bleeding. Age, gastric antrum, and duodenal bulb lesions may be associated with the failure of initial hemostasis by endoscopic hemoclip. PMID- 16340630 TI - Follow-up care after a diagnosis of Helicobacter pylori infection in an Asian immigrant cohort. AB - GOAL: To study the rate at which Helicobacter pylori infection is treated in an immigrant cohort after diagnosis by esophagogastroduodenoscopy (EGD). BACKGROUND: Gastric cancer is the second leading cause of cancer death worldwide, and is especially prevalent in East Asia; immigrants from this part of the world remain at higher risk. Infection with H. pylori is a known risk factor for gastric cancer. There have been no studies of completion of H. pylori treatment in immigrant populations. STUDY: Prospective cohort study of East Asian immigrants diagnosed with H. pylori infection who underwent EGD in a gastric cancer screening protocol. Our primary outcome was self-report or chart evidence of completion of treatment of H. pylori. RESULTS: Sixty-eight of the 126 participants (54%) tested positive for H. pylori infection on EGD. Forty-nine (72%) were seen for a follow-up visit at one of the clinics involved in the study. According to clinic records, 39 of these 49 participants (57% of all H. pylori-positive participants) were prescribed treatment. Only 31 participants (46%) completed treatment. Of possible explanatory factors, only having a "regular doctor" was significantly associated with treatment completion (odds ratio=5.6; 95% confidence interval, 1.2-25.0). CONCLUSIONS: In a sample of Asian immigrants, the rate of treatment of H. pylori infection, a potentially modifiable risk factor, was lower than expected. Having a "regular doctor" appeared to increase the likelihood of receiving appropriate follow-up care. PMID- 16340631 TI - Thyroid disorders in Brazilian patients with celiac disease. AB - INTRODUCTION/AIM: Patients with celiac disease (CD) can develop a gluten related autoimmune disorder that affects not only the small intestine but other tissues as well. An increased prevalence of autoimmune diseases has been reported, particularly autoimmune thyroiditis. The aim of this study was to characterize thyroid disorders in patients with CD. PATIENTS/METHODS: Fifty-two patients with CD (43 female, 9 male; mean age, 41.1 years) were studied. Nine were on a gluten free diet (GFD). They were divided into four groups: Group 1, without thyroid involvement (n=30); Groups 2A-C, with thyroid involvement (n=22); Group 2A, subclinical hypothyroidism (n=11); Group 2B, clinical hypothyroidism (n=10); and Group 2C, other thyroid disorders (n=1). CD was confirmed by serologic and histologic criteria. Thyroid involvement was detected by measurement of thyroid stimulating hormone (TSH) and anti-thyroperoxidase antibodies (anti-TPO). RESULTS: Increased levels of TSH and/or anti-TPO levels were detected in Groups 2A (21.1%) and 2B (19.2%). The patients of Group 2B presented clinical symptoms of hypothyroidism before the diagnosis of CD, and 5 of these patients were receiving levothyroxine. One woman (Group 2C; 1.92%) had a medullary carcinoma. There was statistical significance between the age when thyroid disease was diagnosed (current age) and the age of CD diagnosis between Groups 1 and 2B. Patients with thyroid involvement presented associated diseases such as diabetes mellitus (2), Down's syndrome (2), ulcerative colitis (1), and dermatitis herpetiformis (2). CONCLUSIONS: Our findings demonstrated an increased prevalence of thyroid disorders (hypothyroidism, 19.2%; and subclinical hypothyroidism, 21.2%), and other associated diseases in celiac patients, even on a GFD, increasing with the age of the patients. Screening for associated diseases is recommended for patients with CD, independent of age at diagnosis or treatment duration. PMID- 16340632 TI - A survey on patient educational needs in irritable bowel syndrome and attitudes toward participation in clinical research. AB - BACKGROUND: The educational needs of patients with irritable bowel syndrome (IBS) are poorly understood and rarely studied. AIM: To determine the educational needs of IBS patients, regarding content, presentation format, and expectations from healthcare providers. METHODS: Fifteen functional GI clinic patients were asked open-ended questions to generate items for a questionnaire addressing the study aim. A total of 104 IBS patients received this questionnaire by mail (42 had declined to participate in a prior IBS study). To assess the frequency of endorsements and importance (on a scale of 1-3) of the items, an index was calculated (frequency of endorsements x mean rating per item, first priority scored 3, third priority scored 1). A higher index indicated greater endorsement based on frequency and rating of response. RESULTS: A total of 29 (28%) subjects (22 willing, 7 unwilling to participate previously in questionnaire research) completed the questionnaire (mean age, 42.6 years; SD, 14.2 years; 19 female, 10 male). The overall low response rate is likely related to the population studied; 40.4% of our study subjects have declined participation in prior research. The response rate of those who have previously agreed to participate was 36%. The typical response profile included: interest in learning disease management (index=1.4) and preference for information presented in person by an M.D. (2.4). Choice of presentation media included magazines (1.9), television (1.5), and Web sites (1.2). Doctors' qualities ranked high related to competency (0.8), allocation of sufficient time (0.7), and listening skills (0.4). Preferred incentives for research participation included a thank you note (0.4), summary of trial results (0.3), and monetary incentives (0.6). CONCLUSIONS: This qualitative study will provide pilot data for a national survey on the educational needs of IBS patients, for use in developing effective patient-centered, educational programs. PMID- 16340633 TI - Histologic study of colonic mucosa in patients with chronic diarrhea and normal colonoscopic findings. AB - BACKGROUND: There are controversies about the importance of biopsies of normal colon mucosa in the investigation of patients with chronic diarrhea. STUDY: Colonic and terminal ileum biopsies of 167 patients were reviewed. In 5 patients, used as controls, colonoscopy was done due to family history of colon cancer. RESULTS: The 5 patients without symptoms had no histologic abnormalities. The histologic findings in 162 patients with chronic diarrhea were as follows: 110 patients (67.9%) with normal histology, microscopic colitis not otherwise specified, and isolated small granulomas; 17 (10.5%) patients had findings of borderline diagnostic significance, including possible collagenous colitis, some features of lymphocytic colitis and melanosis coli; and 35 (21.6%) patients, with diagnostic significant histologic findings as collagenous colitis, lymphocytic colitis, minimal change microscopic colitis, eosinophilic colitis, pericrypt eosinophilic enterocolitis, intestinal spirochetosis, schistosomiasis, and Crohn's disease. Of the 52 patients with either borderline or significant diagnostic abnormalities, in 8 (15.4%) the diagnosis was done only with a proximal study (ascending, transverse, or descending colons). CONCLUSIONS: Histologic lesions of possible diagnostic value could exist in 32.1% of chronic diarrhea patients with normal colonoscopy, which can justify, in certain cases, mucosa biopsies, which might contribute for a more precise etiologic diagnosis; also, the distribution of these histologic changes has pointed out the importance of having all colon segments biopsied. PMID- 16340634 TI - Continuation of antibiotics is associated with failure of metronidazole for Clostridium difficile-associated diarrhea. AB - BACKGROUND: Metronidazole is first-line therapy for C. difficile-associated diarrhea primarily because of its low cost relative to vancomycin. Currently, it is unknown which patients will fail metronidazole therapy. Our goal was to prospectively evaluate risk factors for metronidazole failure. STUDY: Included patients had symptomatic C. difficile-associated diarrhea, either mild or severe. Once enrolled, detailed baseline data were gathered. All interviews were performed daily while the patient was in the hospital for up to 14 days. If discharged prior to 14 days, the patient received a follow-up phone call on day 5 and day 14. Enrolled patients were given a daily stool survey to complete. RESULTS: We enrolled 27 patients with C. difficile-associated diarrhea. All patients (10 of 10) who had their offending antibiotic(s) discontinued had symptomatic resolution of diarrhea by day 14 of metronidazole treatment. Conversely, 59% (10 of 17) of patients who remained on antibiotics during treatment had symptomatic resolution by day 14 (P=0.02). The risk ratio for treatment failure was 2.0 (95% confidence interval, 1.29-3.10) in patients who remained on antibiotics. In our treatment group, there would be one additional metronidazole treatment success for every 2.4 patients who discontinued antibiotics. CONCLUSION: Patients who remain on antibiotics while undergoing treatment of C. difficile-associated diarrhea have a high likelihood of treatment failure with metronidazole. PMID- 16340636 TI - Survey of primary physicians' knowledge of colorectal cancer screening. AB - OBJECTIVE: Colorectal cancer (CRC) is the most common of the gastrointestinal cancers in Israel. The low rate of patient compliance to a recent CRC screening program of Clalit Health Services, a major health management organization in Israel, prompted the present survey of primary care physicians' knowledge and practices regarding CRC screening. METHODS: A 23-item questionnaire, formulated according to the policy of the Israel Ministry of Health and the recommendations of the American Gastroenterological Association, was distributed to 150 primary physicians of Clalit Health Services. The relative ratio of correct to incorrect answers was calculated for every question and by professional group. RESULTS: The response rate was 89%. Total score (out of a maximum 23) was 20.68 for family physicians (experts in family medicine), 17.79 for experts in other fields, and 17.82 for general practitioners (average, 0.90, 0.77, and 0.77, respectively). The score for the family physicians was significantly higher than for the other two groups (P=0.0070). Clustering items by specific issues yielded significantly better scores for the family physicians in four areas: screening (P=0.0164), appropriate test for high-risk population in Israel (P=0.0012), definition of average-risk population (P=0.0012), and CRC symptoms (P=0.0108). A low level of knowledge on the definition of the high-risk population was noted in all three groups. CONCLUSIONS: Experts in family medicine in Israel have significantly greater knowledge of most issues of CRC than primary care experts in other fields and general practitioners, although all three groups lack knowledge on the definition of the high-risk population. Continuing physician education should focus on these areas. PMID- 16340635 TI - Treatment with infliximab is associated with increased markers of bone formation in patients with Crohn's disease. AB - OBJECTIVE: Osteoporosis is a common complication of Crohn's disease (CD). Glucocorticoid use and detrimental effects of inflammatory cytokines including tumor necrosis factor-alpha (TNF-alpha) can lead to osteoporosis. The aim of this study was to assess the ability of treatment with the TNF-alpha antagonist infliximab to increase bone formation as measured by surrogate markers of bone turnover in patients with active CD. METHODS: Sera from 38 prospectively enrolled CD patients were examined for levels of bone alkaline phosphatase (BAP), N telopeptide of type I collagen (NTX), immunoreactive parathyroid hormone (iPTH), calcium, and pro-inflammatory cytokines at baseline and 4 weeks following infliximab infusion. Crohn's Disease Activity Index (CDAI), Inflammatory Bowel Disease Questionnaire (IBDQ), and glucocorticoid dose also were collected. RESULTS: In this cohort, CDAI and IBDQ scores were significantly improved at week 4 (P<0.001). Infliximab therapy was associated with an increase in BAP, a marker of bone formation (P=0.010), whereas NTX, a marker of bone resorption, was not increased (P=0.801). Among 22 patients who were taking glucocorticoids, mean glucocorticoid dose decreased 36% (P<0.001; -7.9 mg). CONCLUSIONS: Treatment with infliximab was associated with increased markers of bone formation (BAP) without increasing bone resorption (NTX). This effect may be due to a beneficial effect of TNF-alpha blockade on bone turnover, a beneficial effect on CD activity resulting in decreased glucocorticoid dose, or both. Studies of longer duration are needed to assess the effect of infliximab on bone mineral density. PMID- 16340637 TI - Liver disease in patients with diabetes mellitus. AB - Diabetes mellitus is a growing health concern in our society. In addition to the well-known cardiovascular, renal, and ophthalmologic complications of diabetes, liver-related complications occur commonly and are often underrecognized. The ensuing article will review the relationship between diabetes mellitus and two common liver diseases: chronic hepatitis C and nonalcoholic fatty liver disease. The association with diabetes and cirrhosis, acute liver failure, hepatocellular carcinoma, and outcomes following orthotopic liver transplantation will also be discussed. PMID- 16340638 TI - Evaluation of intrapulmonary vascular dilatations with high-resolution computed thorax tomography in patients with hepatopulmonary syndrome. AB - GOALS: We aimed to determine the role of thorax high-resolution computed tomography (HRCT) in demonstrating the pulmonary vasodilatation in patients with hepatopulmonary syndrome (HPS). BACKGROUND: Traditionally, the presence of intrapulmonary vascular dilatations can be detected by using one of the three diagnostic modalities: contrast-enhanced echocardiography, technetium 99 m labeled macroaggregated albumin scan, and pulmonary angiography. STUDY: The study group included 10 patients with HPS (Group 1), 12 patients with normoxemic cirrhosis (Group 2), and 12 healthy controls (Group 3). All of the subjects underwent conventional and HRCT of thorax. The diameters of pulmonary trunk, main pulmonary arteries, and right lower lobe basal segmental arteries were measured. The ratios of right lower lobe basal segmental pulmonary artery to bronchus diameter were calculated. RESULTS: The mean diameters of the main pulmonary trunk, right and left main pulmonary arteries were not different between the groups. Mean diameters of right lower lobe basal segmental pulmonary arteries were significantly higher in Group 1 compared with Group 2 (P=0.01) and Group 3 (P=0.002). Mean right lower lobe basal segmental pulmonary artery to bronchus ratios were significantly higher in Group 1 compared with Group 2 (P=0.03) and Group 3 (P<0.001). Group 2 had significantly higher pulmonary artery to bronchus ratios than Group 3 (P<0.001). CONCLUSIONS: Thorax HRCT may be helpful in the diagnosis of HPS by demonstrating the dilated peripheral pulmonary vessels or increased pulmonary artery to bronchus ratios in patients with liver disease and hypoxemia. PMID- 16340639 TI - Bleeding colonic ulcer and eosinophilic colitis: a rare complication of nonsteroidal anti-inflammatory drugs. PMID- 16340640 TI - Neurofibroma in the colon. PMID- 16340641 TI - Octreotide for unresectable hepatocellular carcinoma: beyond the first sight. PMID- 16340642 TI - The general practitioner's management of patients with a new diagnosis of irritable bowel syndrome. PMID- 16340643 TI - Hemolytic anemia during 24 weeks of ribavirin and interferon-alpha2b combination therapy does not influence the cardiac function of patients with viral hepatitis C. PMID- 16340644 TI - Dysphagia in aging. PMID- 16340646 TI - Magnetic resonance imaging findings in substance abuse: alcohol and alcoholism and syndromes associated with alcohol abuse. AB - Alcohol abuse is common among the population and results in significant diseases that shorten life span. Ethanol may result in chronic brain changes such as atrophy but may also result in neurologic disease that may be acute or chronic and sometimes life threatening. Accompanying vitamin deficiencies may lead to Wernicke's encephalopathy and changes in serum osmosis may lead to several acute demyelinating disorders. In addition, pregnant women who consume alcohol place their babies at high risk for the fetal alcohol syndrome. In this article we review these disorders and emphasize their imaging features. PMID- 16340647 TI - Neuroimaging in drug and substance abuse part I: cocaine, cannabis, and ecstasy. AB - Multiple neurological complications of cocaine abuse have been described including both ischemic and hemorrhagic cerebrovascular events, atrophy in the case of chronic abuse, and an increase in incidence of congenital malformations in the setting of maternal use. The abuse of cannabis may cause a number of neurovascular changes that, in turn, may result in ischemic events, however, a direct connection between these has not been fully established. The use of MDMA (ecstasy), a popular recreational drug, has been related to ischemic and hemorrhagic cerebrovascular events, as well as atrophy. Neuroimaging studies are vital in the assessment of the extent of neurologic damage in these patients. PMID- 16340648 TI - Neuroimaging in drug and substance abuse part II: opioids and solvents. AB - The central nervous system is one of the primary targets for the detrimental effects of drugs of abuse. Diagnostic imaging, especially MRI, plays an important role in the detection of complications associated with drug abuse. We present the imaging findings associated with the abuse of opioids and other morphine derivatives, as well, as solvents. Of the morphine derivatives, heroin is the most commonly abused. Several CNS pathologic effects have been described in association with its abuse. These include neurovascular complications such as microvascular ischemic changes or ischemic stroke. A rare form of leukoencephalopathy has been described in those abusers who inhale heroin vapors. Other neurologic complications include atrophy and various infectious processes. Solvent inhalation is a common practice among adolescents and young adults secondary to its ease of access and low cost. The most important component of industrial solvents is toluene. Complications of toluene abuse may be either acute, showing no neuroradiological changes, or chronic, characterized by cerebral and cerebellar demyelination as well as atrophy. PMID- 16340649 TI - Functional magnetic resonance and spectroscopy in drug and substance abuse. AB - OBJECTIVES: Functional magnetic resonance imaging (fMRI) and proton magnetic resonance spectroscopy (1H-MRS) were utilized to evaluate functional and metabolic brain changes in drug abusers. METHODS: Functional neuroimaging studies in alcohol, cocaine, cigarette, and cannabis abusers were reviewed in the medical literature from 2000 to 2005. RESULTS: Brain activation with functional magnetic resonance imaging is different in drug abusers and may show more or less activation according to the task and gender. Lower concentrations of N acetylaspartate and higher creatine are usually seen with 1H-MRS in the brains of cigarette smokers, people who are cocaine dependent, and alcohol abusers. CONCLUSIONS: Neural substrates of drug addition, craving, drug interaction, individual abstinence, and drug withdrawal can be studied with functional magnetic resonance imaging and proton magnetic resonance spectroscopy. PMID- 16340650 TI - PET and SPECT in drug and substance abuse. AB - The use of imaging in the evaluation of the brain under the influence of drugs has evolved into a functional assessment of its processes. It has been shown that SPECT and PET imaging studies demonstrate selected areas that are affected by multiple drugs among substance abusers. This paper illustrates examples of different drugs and how their effect on the brain may be portrayed by imaging applications. The developing "road maps" that we have obtained possess multidimensional applications. They can be useful to confirm specific diseases, clarify diagnosis wherein the clinical picture is not observed, aid in the development of new treatment evaluations and also provide researchers a better insight in the detection and functionality of dementia and specific mental illnesses processes. PMID- 16340653 TI - Impact of nutrition on ageing and disease. AB - PURPOSE OF REVIEW: The globe is suffering a tsunami of chronic diseases, affecting especially the elderly and those with a dysfunctioning immune system. The fundamental principles of optimal health and optimal ageing are abstaining from smoking, modest alcohol consumption, regular physical exercise and a diet rich in fish and plants and low in condensed calories, sugar and dairy products. RECENT FINDINGS: Dietary supply and production of advanced glycation end products leads to the accumulation of these products in the tissues and is strongly associated with ageing of the vascular endothelium, nervous system, eyes and other vital organs. Telomeres, which are not involved in DNA repair, remain unrepaired and loose with time. A decline in innate and acquired immunity is seen with increasing age and maintenance of low basal immune activity (degree of inflammation) seems important for health and longevity: 'people who are predisposed to weak inflammatory activity may live longer'. SUMMARY: Supplementation with vitamins has little effect on ageing/prevention of chronic diseases, but antiinflammatory molecules like polyphenols are more effective, especially when combined with reduced intake of calorie-condensed foods. The effect of probiotics on ageing needs further exploration. The effects of caloric restriction, proven effective in other species to control aging and prolong lifespan, have not been fully explored in humans. PMID- 16340654 TI - Dietary cholesterol provided by eggs and plasma lipoproteins in healthy populations. AB - PURPOSE OF REVIEW: Extensive research has not clearly established a link between egg consumption and risk for coronary heart disease. The effects of egg intake on plasma lipids and low-density lipoprotein (LDL) atherogenicity in healthy populations need to be addressed. RECENT FINDINGS: The lack of connection between heart disease and egg intake could partially be explained by the fact that dietary cholesterol increases the concentrations of both circulating LDL and high density lipoprotein (HDL) cholesterol in those individuals who experience an increase in plasma cholesterol following egg consumption (hyperresponders). It is also important to note that 70% of the population experiences a mild increase or no alterations in plasma cholesterol concentrations when challenged with high amounts of dietary cholesterol (hyporesponders). Egg intake has been shown to promote the formation of large LDL, in addition to shifting individuals from the LDL pattern B to pattern A, which is less atherogenic. Eggs are also good sources of antioxidants known to protect the eye; therefore, increased plasma concentrations of lutein and zeaxanthin in individuals consuming eggs are also of interest, especially in those populations susceptible to developing macular degeneration and eye cataracts. SUMMARY: For these reasons, dietary recommendations aimed at restricting egg consumption should not be generalized to include all individuals. We need to acknowledge that diverse healthy populations experience no risk in developing coronary heart disease by increasing their intake of cholesterol but, in contrast, they may have multiple beneficial effects by the inclusion of eggs in their regular diet. PMID- 16340655 TI - Calcium, vitamin D and involutional osteoporosis. AB - PURPOSE OF REVIEW: Previous studies suggest that combined calcium and vitamin D supplementation decreases the risk of fractures in older people, particularly those living in care homes, but trials of vitamin D alone in fracture prevention have generated inconsistent results. This review examines the physiological functions of calcium and vitamin D, and the contrasting views of what constitutes an adequate dietary calcium intake and vitamin D sufficiency in adults, and highlights the results of recent large studies of calcium and vitamin D supplementation. RECENT FINDINGS: The RECORD study shows that calcium (1000 mg/day) and vitamin D (800 IU/day), either alone or in combination, are ineffective in the secondary prevention of osteoporotic fractures in older men and women living in the community. The Northern and Yorkshire Study also suggests that calcium (1000 mg/day) and vitamin D (800 IU/day) are of no benefit in the primary prevention of fractures in community-dwelling older women. Furthermore, the Wessex study demonstrated no reduction in fractures in older people living in the community treated with annual IM injections of vitamin D (300 000 IU). SUMMARY: The latest studies highlight that vitamin D, either alone or in combination with calcium supplementation, is ineffective in the primary or secondary prevention of fractures in community-dwelling older people. In contrast, calcium and vitamin D supplementation prevents fractures in institutionalized elderly people, who commonly have vitamin D deficiency and secondary hyperparathyroidism. PMID- 16340656 TI - Nutrition and cardiac cachexia. AB - PURPOSE OF REVIEW: Congestive heart failure is a leading cause of morbidity and mortality, especially in older persons. In advanced stages of the disease, congestive heart failure can be associated with serious complications such as cardiac cachexia (defined here as weight loss of more than 6% in 6 months). This review will discuss recent insights into the pathophysiology, anthropometric predictors and potential management of cardiac cachexia. RECENT FINDINGS: Cardiac cachexia and the associated progressive weight loss are sometimes overlooked by care providers. A delay in diagnosis often results in further loss of vital tissues, progressive weakness, fall-related injuries and potentially long-term care institutionalization and/or death. Emerging data suggest that congestive heart failure is a dynamic disorder of many organ systems, including the myocardial, neurohormonal, immune, vascular, gastrointestinal, renal and musculoskeletal systems. It is becoming more widely appreciated that it is the deterioration of this interactive multisystem complex that results in the systemic inflammation and progressive wasting and atrophy of muscle and other organ tissues, which is the hallmark of cardiac cachexia. SUMMARY: Cardiac cachexia in congestive heart failure patients may be associated with a low level of physical activity. A high systemic inflammatory state is another marker of cardiac cachexia. Prudent anti-inflammatory nutrition, dietary supplements and exercise can serve to ameliorate and/or potentially prevent progressive wasting. A better understanding of factors contributing to the development of cardiac cachexia will enable us to design preventive strategies and provide improved care for individuals with this debilitating condition. PMID- 16340657 TI - Bibliography. Current world literature. Ageing: biology and nutrition. PMID- 16340659 TI - Podocyte injury and targeting therapy: an update. AB - PURPOSE OF REVIEW: Podocyte injury is a central event in the development of glomerulosclerosis. This review highlights contributions from the past year to our understanding of mechanisms of podocyte injury and implications for potential treatment strategies of glomerular disease. RECENT FINDINGS: Rearrangement of the actin cytoskeleton, the backbone linking the slit diaphragm, apical domain and sole plate, serves as a common denominator during foot process effacement. Reports on the role of synaptopodin and CDK5 on actin dynamics as well as cathepsin L and B7.1 in subsequent cell migration have expanded our understanding of the podocyte response to injury. Mounting evidence supports an expanding role of the slit diaphragm in signal transduction to mediate downstream cellular responses, including prosurvival effects of the integral proteins nephrin and CD2AP. The discovery that TRPC6 localizes to the slit diaphragm and identification of specific mutations of the transport channel in kindreds of familial focal segmental glomerulosclerosis implicate a causal role for aberrant calcium signaling in podocyte injury. Disruption of the dystroglycan complex, which anchors the podocyte to the underlying basement membrane, in states of foot process effacement may have implications for the recent finding of viable podocytes in the urine in glomerular disease. SUMMARY: The resurgence of research in podocyte biology over the past decade underscores the importance of this unique cell in preserving glomerular structure and function. A greater understanding of the complex signaling mechanisms governing podocyte biology in health and disease will ultimately lead to novel therapeutic avenues for treating disorders of the podocyte. PMID- 16340660 TI - The tissue renin-angiotensin system and intracellular signalling. AB - PURPOSE OF REVIEW: The renin-angiotensin system is not what it was, or for that matter not necessarily where we thought it should be. For example, there is a novel angiotensin I-metabolizing enzyme that generates angiotensin 1-7 rather than angiotensin II. Moreover, we are slowly realizing the importance of local rather than circulating angiotensin II. RECENT FINDINGS: Rather than concentrating on the systemic renin-angiotensin system, recent work has concentrated on elucidating the consequences of increasing angiotensin II production within specific organs, such as the heart and vasculature, as well as in the pancreas and in adipose tissue. Inhibition of angiotensin II production either using angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers not only reverses remodelling but also increases tissue insulin sensitivity. Targeting the renin-angiotensin system clinically delays the onset of type 2 diabetes, but the mechanisms involved are not clearly understood. Moreover, at least one other angiotensin-converting enzyme homologue (ACE2) plays a significant role in the regulation of heart and kidney function, and as it generates angiotensin 1-7 from angiotensin I, it is proposed to counteract the detrimental effects associated with the activation of the classic renin angiotensin system. SUMMARY: There is a need to re-evaluate the role(s) played by the molecular components of the "extended" local renin-angiotensin system and their role in vascular disease and type 2 diabetes. PMID- 16340661 TI - The natriuretic peptide system: kidney and cardiovascular effects. AB - PURPOSE OF REVIEW: The natriuretic peptide (NP) system is primarily an endocrine system that maintains fluid and pressure homeostasis by modulating cardiac and renal function. The physiologic functions of the NP system in healthy humans and in patients with cardiovascular disease are not fully understood. NP levels are elevated in patients with heart failure (HF) and other cardiac diseases; measurement of NPs may be used in the clinical setting to aid diagnosis and prognosis. In addition, synthetic NPs such as nesiritide are available for use in management of patients with acutely decompensated congestive HF. RECENT FINDINGS: Not only do NPs modulate volume and pressure homeostasis, but they also exert important anti-proliferative, anti-fibrotic effects in the heart. Thus, NPs may prove useful for prevention of remodeling after myocardial infarction and in advanced HF. BNP is emerging as an important biomarker in patients with HF and other cardiovascular diseases, such as pulmonary hypertension and atherosclerotic vascular disease. Elevated NP levels may serve as an early warning system to help to identify patients at high risk for cardiac events. Recombinant human ANP (carperitide) and BNP (nesiritide) are useful for management of acutely decompensated HF; these drugs are also being investigated for myocardial and renal protection in the setting of cardiac surgery and for prevention of cardiac remodeling. SUMMARY: The clinical application of NPs is expanding rapidly. Recent basic science and clinical research findings continue to improve our understanding of the NP system and guide use of ANP and BNP as biomarkers and as therapeutic agents. PMID- 16340662 TI - Cardioprotective mechanisms of the kallikrein-kinin system in diabetic cardiopathy. AB - PURPOSE OF REVIEW: Multiple pathogenic mechanisms contribute to the development of diabetic cardiopathy, including intramyocardial inflammation, cardiac fibrosis, abnormal intracellular Ca handling, microangiopathy and endothelial dysfunction. Moreover, the cardiac kallikrein-kinin system is thought to be altered under diabetic conditions and an improvement of this peptide system, e.g. by gene therapeutic approaches, has also been associated with an amelioration of the diabetic heart. In this review, we will discuss the hypothesis that the stimulation of the kallikrein-kinin system could be a promising target for the treatment of diabetic cardiopathy. RECENT FINDINGS: The kallikrein-kinin system has cardioprotective properties, which may be particularly important under diabetic conditions. For example, its potential for endothelium-dependent vasodilation, and for improvement of glucose transport and utilization, make bradykinin an important mediator for reducing the consequences of diabetes related oxidative stress on both the myocardium and vessels. SUMMARY: The different synergistic cardioprotective effects of the kallikrein-kinin system in the diabetic heart suggest that the stimulation of the kallikrein-kinin system might open new avenues for the treatment of diabetic cardiopathy. PMID- 16340664 TI - The renal medullary endothelin system in control of sodium and water excretion and systemic blood pressure. AB - PURPOSE OF REVIEW: Endothelin-1 is a multifunctional peptide that is produced by the kidney and may regulate a variety of renal functions. This review discusses recent developments in understanding the role of the medullary endothelin-1 system in regulating renal salt and water excretion and systemic blood pressure. RECENT FINDINGS: The renal medulla is the major site of endothelin-1 synthesis and receptor expression in the kidney. Endothelin-1 in vitro can inhibit sodium or water transport in the collecting duct and thick ascending limb through autocrine pathways. Endothelin-1 also can increase medullary blood flow. These effects of endothelin-1 are partially mediated by nitric oxide and cyclooxygenase metabolites which are produced by most medullary cells. Mice with collecting duct specific knockout of the endothelin-1 gene have impaired sodium excretion in response to sodium loading and have hypertension which worsens with high salt intake. The mice also have heightened sensitivity to vasopressin and decreased ability to excrete an acute water load. Mice with collecting duct-specific endothelin A receptor knockout have normal blood pressure and sodium excretion, but have reduced vasopressin responsiveness. Medullary endothelin-1 content is reduced in many forms of experimental hypertension. SUMMARY: Medullary endothelin 1 regulates renal sodium and water transport and medullary blood flow. In particular, the medullary collecting duct is important in this process, but the medullary endothelin system involves complex interactions, through autocrine and paracrine pathways, between most cell types in the region. Medullary endothelin-1 is fundamentally important in physiologic regulation of renal sodium and water excretion and maintenance of normal systemic blood pressure. PMID- 16340663 TI - Hormonal and cytokine effects of uric acid. AB - PURPOSE OF REVIEW: Current evidence supports the role of soluble uric acid as a true mediator of injury, exerting its effects through the induction of growth factors, cytokines, hormones and autacoids. In the present review, we summarize recent studies on the mechanisms involved in the uric acid deleterious effects. RECENT FINDINGS: Although uric acid is considered an antioxidant in plasma, recent clinical and epidemiological studies have found that hyperuricemia is associated with mortality and development of hypertension, cardiovascular and chronic renal diseases. Experimental studies suggest that uric acid induce its detrimental effects at the cellular level entering to vascular smooth muscle cells (VSMC) via an organic anion transport system, and followed by the activation of specific MAP kinases, nuclear transcription factors, with stimulation of COX-2, PDGF A and C chain, PDGF alpha receptor, and various inflammatory mediators, including C-reactive protein and monocyte chemoattractant protein-1. Physiologically, these effects translate into a rise of arterial pressure, VSMC hypertrophy, tubulointerstitial infiltration and glomerular hypertension in the setting of renal vasoconstriction. Uric acid also promotes endothelial dysfunction through inactivation of NO and arresting the proliferation of endothelial cells. Thus, arteriosclerosis induced by hyperuricemia may be a novel mechanism for the development of essential hypertension. SUMMARY: Soluble uric acid has important biologic roles. While it acts as an antioxidant, there is also evidence that uric acid has pro inflammatory and proliferative effects on VSMC, and causes dysfunction of endothelial cells. These cellular mechanisms may translate into why uric acid is associated with renal and cardiovascular disease. PMID- 16340665 TI - Systolic blood pressure as the trigger for the renal myogenic response: protective or autoregulatory? AB - PURPOSE OF REVIEW: The ability of the kidney to autoregulate renal blood flow and glomerular filtration rate has long been viewed as existing to prevent fluctuations in blood pressure from causing parallel fluctuations in renal function and distal delivery of filtrate. This review, however, points out that the primary consequence of the loss of this autoregulatory capacity is not a disturbance in volume regulation, but rather an increased susceptibility to hypertensive injury. Moreover, the kinetic requirements for renal protection indicate that current views of dynamic autoregulation cannot explain how the kidney is normally protected against acute elevations in systolic blood pressure. RECENT FINDINGS: Recent findings suggest that the kinetics of the myogenic mechanism of the afferent arteriole are uniquely suited to protect against acute elevations in the systolic blood pressure, in that this vessel not only senses this rapidly oscillating blood pressure component, but that its response is exclusively dependent on this signal. SUMMARY: These new findings are consistent with recent data indicating that it is the systolic blood pressure elevations that most closely correlate with target organ damage. The fact that the myogenic mechanism is also a necessary component of renal autoregulation may explain the strong linkage between autoregulatory impairment and increased susceptibility to hypertensive injury. PMID- 16340666 TI - Angiotensin II compartmentalization within the kidney: effects of salt diet and blood pressure alterations. AB - PURPOSE OF REVIEW: All components of the renin-angiotensin-aldosterone system are present within the kidney. Renin, renin receptor, angiotensinogen and angiotensin AT1 and AT2 receptor and aldosterone synthase messenger RNA and protein are present in close proximity to the renal vasculature and tubules. The interaction between the different components of the renin-angiotensin-aldosterone system determines the level of activity of this system and in turn may influence the regulation of blood pressure and renal sodium handling. RECENT FINDINGS: Angiotensin through the stimulation of its subtype AT2 receptor regulates sodium excretion, renin synthesis and secretion. Aldosterone synthase mRNA and protein are expressed in glomeruli, renal vasculature and tubules, and are regulated by angiotensin AT1 receptor, diabetes and salt. Although aldosterone is known to influence renal tubular channels with the subsequent enhancement of sodium reabsorption, it is not clear if the renally produced aldosterone also influences renal sodium handling or blood pressure regulation. In addition, angiotensin II influences kidney function and structure through the stimulation of renal inflammation. New data suggest that the renal AT1 receptor plays an important role in the determination of blood pressure levels, and this effect is unique and non-redundant in the actions of extrarenal AT1 receptors. SUMMARY: The finding of new functions and components of the renin-angiotensin-aldosterone system clearly adds new knowledge to our understanding of how angiotensin II influences the kidney and blood pressure. PMID- 16340667 TI - Effect of salt intake on progression of chronic kidney disease. AB - PURPOSE OF REVIEW: The attempt of this review is to bring into focus the potential role of dietary salt intake in progression of chronic kidney disease. RECENT FINDINGS: Ongoing work has elucidated a role for dietary salt intake in modulating intrarenal production of transforming growth factor-beta1. The mechanism is independent of angiotensin II and systemic blood pressure and involves activation of vascular endothelium by dietary salt intake with release of this growth factor. In this model, transforming growth factor-beta1 serves an autacoid function by stimulating nitric oxide production by the endothelium. In turn, endothelium-derived nitric oxide modulates production of this growth factor. The model further predicts that individuals who have lost the requisite endothelial cell flexibility to adapt to this environmental stress (a high salt diet) are potentially at increased risk of developing end-organ damage from excess salt intake. Animal and human studies are presented to support this working hypothesis. SUMMARY: Overproduction of transforming growth factor-beta1 permits excess biological activity of this important fibrogenic growth factor with subsequent development or acceleration of vascular and kidney damage. In patients with diseases whose pathogenesis is related to excess production of transforming growth factor-beta1, such as chronic allograft nephropathy and diabetic nephropathy, increased salt intake may hasten loss of function, particularly if nitric oxide production does not increase. The role that endothelial cell plasticity plays in altering vascular tone and renal function, especially in response to changes in dietary salt intake, should be examined further in chronic kidney disease. PMID- 16340668 TI - Analysis of cardiovascular disease and kidney outcomes in multidisciplinary chronic kidney disease clinics: complex disease requires complex care models. AB - PURPOSE OF REVIEW: Chronic kidney disease is recognized as being highly prevalent in the population, and associated with morbidity and mortality relative to the general population. The complexity of patients and the multiplicity of interventions required to maintain health has forced clinicians to develop different models of healthcare delivery. This publication reviews the current literature on specific interventions to reduce progression of chronic kidney disease and cardiovascular disease, and studies the examination of outcomes of patients exposed to different healthcare delivery models. Specifically we examine the rationale and outcomes of those seen in multidisciplinary clinics. RECENT FINDINGS: Current evidence supports the use of rennin-angiotensin system blockers, reduction of blood pressure and proteinuria and phosphate control. Additional less robust studies support the need for attention to anemia, hyperparathryoidism, and other more "kidney specific" risk factors. The attendance of identified chronic kidney disease patients at multidisciplinary clinics appears to improve survival once dialysis is started. Despite aggressive management, not all patients are able to meet clinical targets associated with improved outcomes. SUMMARY: The recognition of the complexity of chronic kidney disease care and the need to develop and test models of care in addition to the single interventions is a challenge for both researchers and clinicians. Current data support the use of multidisciplinary clinics in improving outcomes of referred patients. Future research will help to refine and define appropriate care models for this growing chronic kidney disease population. PMID- 16340669 TI - The vascular wall as a regulator of tissue oxygenation. AB - PURPOSE OF REVIEW: The development of the phosphorescence quenching oxygen measurement technique has allowed for a simultaneous measurement of intra and perivascular partial pressure oxygen along arteriolar vessels in vivo. Mapping the microvascular distribution and oxygen gradients across the vascular walls using this high-resolution technique reveals the existence of large radial gradients between the vasculature and the tissue, with concomitant longitudinal oxygen loss. Mass balance analysis along vessel segments indicates that the vascular wall has a high rate of oxygen consumption. This review presents the current status of in-vivo studies on the partitioning of oxygen between blood, the vascular wall and the surrounding tissue, thereby positioning an oxygen sink between blood and tissue regulating oxygen release. RECENT FINDINGS: Induced vasoactivity (vasoconstriction and vasodilation) has been shown to modulate oxygen consumption of the vascular wall and directly affect the portion of oxygen available to the tissue. Inhibition of the endothelial layer of the vessel wall resulted in a decrease in the oxygen gradient across the vessel. SUMMARY: The vascular wall is a sink for oxygen. The modulation of vessel wall oxygen consumption can substantially impact the amount of oxygen released to the tissue. PMID- 16340670 TI - Oxidative stress and nitric oxide in kidney function. AB - PURPOSE OF REVIEW: Nitric oxide is a potent, endogenous vasodilator that regulates systemic blood pressure and renal function, among other functions. The bioactivity of nitric oxide is reduced by superoxide, a major reactive oxygen species. Overproduction of superoxide and other related reactive oxygen species resulting in oxidative stress reduces the biological effects of nitric oxide. Though both of these highly reactive species have distinct roles in other pathways, their interaction is emerging as a major regulatory factor in normal and pathological renal function. The purpose of this review is to highlight the recent studies on oxidative stress and nitric oxide in the kidney, focusing on their interaction in normal and pathological conditions. RECENT FINDINGS: Studies have focused on pro-oxidant pathways and nitric oxide defense systems in normal and pathological conditions. The oxidant potential of uncoupled nitric oxide synthases is gaining interest as a pro-oxidant system. Both animal and clinical studies have attempted to identify strategies to intervene at various stages of the oxidant-nitric oxide pathways to improve function during renal failure. SUMMARY: Several new approaches and provocative findings have emerged over the last year. A regulatory role for nitric oxide in the control of the renal microcirculation and as a participant in tubule function is further described. New information of the cause and possible prevention of acute and chronic renal failure has also been produced in the last year. These advances demonstrate the value of research in the normal and pathological roles of oxidative stress and nitric oxide in the kidney. PMID- 16340671 TI - Bibliography. Current world literature. Hormones, autacoids, neurotransmitters and growth factors. PMID- 16340674 TI - Values expressed through intergenerational family food and nutrition management systems among African American women. AB - This grounded theory investigation aimed to understand intergenerational family roles and the food management strategies of African American women from a social ecological perspective. Thirty women from 10 low/moderate-income 3-generation urban families participated in interviews covering roles, health, nutrition, and food management strategies. Four dynamic family systems for managing food and nutrition emerged from qualitative data analysis. Participants expressed values of responsibility, social connections, caretaking, reward, and equal opportunity, and fulfilling responsibilities for family care, connections, and finances. These values and systems provide a basis for culturally appropriate, interpersonal level nutrition interventions among African American women that build on family structures, needs, and resources. PMID- 16340675 TI - Outcome expectations, barriers, and strategies for healthful eating: a perspective from adolescents from low-income families. AB - To better understand outcome expectations, barriers, and strategies for healthful eating, 5 structured focus groups were conducted with adolescents from low-income families (n = 48; 83% Black students; 10-14 years old). The focus group questions dealt with healthful eating in different settings: home, school, and other eating establishments. Results suggested that healthful eating is important to this population, but perceived barriers prevent the consistent consumption of healthful foods. Strategies suggested by participants to increase healthful eating (eg, parents and schools making healthy foods that look and taste good more available) should be considered when developing nutrition interventions for adolescents from low-income families. PMID- 16340676 TI - Public health perspectives on the family: an ecological approach to promoting health in the family and community. AB - The family, as a setting of practice, is increasingly recognized as critical to health promotion. A better understanding of the nature and process through which families take an active part in their own health can serve as the basis for designing and linking health interventions with public health programs. The integrating function of the family, viewed through an ecological context, makes it an effective entry point and central focus in health promotion. PMID- 16340677 TI - Changing the home nutrition environment: effects of a nutrition and media literacy pilot intervention. AB - The specific aim for this pilot study was to evaluate the effectiveness of a nutrition and media literacy intervention targeting elementary students and their parents. The purpose of the intervention was to increase child fruit and vegetables (FV) consumption and change the home nutrition environment (measured with FV availability and accessibility and parental social support). During the intervention, students learned about nutrition, the role media plays in shaping values concerning nutrition, and developed a media campaign for their parents. A quasi-experimental research design was used to evaluate the effectiveness of the intervention. The media intervention was effective in changing the home environment. PMID- 16340678 TI - A socioecological analysis of the determinants of national public health nutrition work force capacity: Australia as a case study. AB - This article uses a socioecological analytical approach to assess the capacity of the public health nutrition work force in Australia as a prelude to work force development strategy planning. It demonstrates how the socioecological model can be used to assess and inform the development of the infrastructure required for effective public health nutrition effort. An interpretive case study method was used involving triangular analysis of quantitative and qualitative data from multiple sources including semistructured interviews with advanced-level practitioners, literature review, a cross-sectional national work force survey, and position description audit and consensus development using a Delphi study. The findings of this analysis indicate that the Australian public health nutrition work force's capacity to effectively address priority nutrition issues is limited by determinants that can be categorized as relating to human resource infrastructure, organizational and policy environments, intelligence access and use, practice improvement and learning systems, and work force preparation. This socioecological analysis supports an intelligence-based focus for work force development effort in Australia and a conceptual framework for work force capacity assessment with potential applications in other countries. PMID- 16340679 TI - Quality tools to improve care and prevent errors. PMID- 16340680 TI - Nurses and the Joint Commission tracer methodology. PMID- 16340681 TI - Staff nurses' use of report card data for quality improvement: first steps. PMID- 16340682 TI - Integrating patient safety into practice. PMID- 16340683 TI - Implementation of patient centeredness to enhance patient safety. PMID- 16340684 TI - Guided Growth Intervention: from novice to expert through a mentoring program. PMID- 16340685 TI - Tobacco ban within a psychiatric hospital. PMID- 16340686 TI - Am I safe here?: improving patients' perceptions of safety in hospitals. AB - Little is known about the current status of hospitalized patients' perceptions of safety or what affects them. This study analyzes safety ratings data of more than 600,000 patients. After describing how patients rate their safety in US acute care hospitals, we delineate the greatest opportunities for improving patient experiences of safety in hospitals. In addition, we offer practical strategies for planning improvements in safety-related elements of care from the patient's perspective. PMID- 16340688 TI - How satisfied are parents supported by nurses with the NIDCAP model of care for their preterm infant? AB - The main purpose of implementing the Newborn Individualized Developmental Care and Assessment Program (NIDCAP) in our neonatal intensive care unit from the perspective of quality of care was to bring about an improvement in the satisfaction of parents. This was measured by means of the NICU-Parent Satisfaction Form and the Nurse Parent Support Tool. Parents were significantly more satisfied with care given according to NIDCAP principles than they were with the traditional care for their premature born babies. PMID- 16340689 TI - The Shift Coupon: an innovative method to monitor adverse events. AB - Adverse events are one method of measuring hospital quality; however, collecting these data is problematic. This article reports findings from a study conducted to collect data about the occurrence of adverse events in the hospital setting using an instrument called the Shift Coupon. The results demonstrated the viability of the Shift Coupon to collect adverse events data and identified the most commonly reported adverse events and nurse-reported causes of adverse events. PMID- 16340690 TI - Quality of life after participation in a rural phase II cardiac rehabilitation program. AB - The purpose of this study was to explore the effectiveness of a cardiac rehabilitation program (phase II) in improving participants' quality of life (QOL). Existing data collected previously by a not-for-profit rural community hospital using the SF-36 tool were analyzed. The secondary data consisted of precardiac and post-cardiac rehabilitation SF-36 scores for 121 phase II participants. All the 8 subscale post scores of the SF-36 were higher when compared to the pre-rehabilitation scores. In this non-controlled trial, results suggest that participation in rural cardiac rehabilitation programs may improve patients' perceptions of QOL and health. PMID- 16340691 TI - BCLS certification of the nursing staff: an evidence-based approach. AB - This article describes the application of the Iowa Model of Evidence-Based Practice to Promote Quality Care to creating standards of practice for Basic Cardiac Life Support certification of the nursing staff at a merged health center in Montreal, Quebec, Canada. Selected outcomes and adopted practice guidelines are presented. PMID- 16340692 TI - A model for quantifying quality initiatives in intensive care. AB - A balance should exist in health care organizations between the focus on cost containment and quality improvement efforts. This is particularly important in relation to critical care in that intensive care units (ICUs) absorb a significant proportion of the hospital's budget. The purpose of this article is to present a model that can be used to quantify the financial value to the institution of quality ICU outcomes achieved as a result of implementation of ventilator bundle initiatives. This one-bed ICU model illustrates the value of improving the care for one patient and cost, quantifying the outcomes that result through a focus on cost savings, revenue enhancement, and cost avoidance. PMID- 16340693 TI - Who is dying in our critical care units? A single center's experience. AB - Are deaths that occur in critical care expected or unexpected? The objective was to illustrate the incidence of deaths in adult critical care units. We also wanted to discover if our patient population fit the norm of those who die in these units in the United States. Are the patients with many comorbid conditions? Could they benefit from advance care planning? A prospective chart review was completed on all deaths that occurred in our 5 critical care units. Most deaths (76.6%) were expected. Patients had an average of 3.3 comorbidities. Greater public knowledge about advance care planning is needed and must include education about the full range of options in end-of-life care. PMID- 16340694 TI - Reducing health disparities by improving quality of care: lessons learned from culturally diverse women. AB - Disparities in health care for culturally diverse populations in the United States lead to poorer health outcomes. This article focuses on decreasing health disparities by improving the quality of care for culturally diverse women. Findings from 3 pilot studies are organized using Fongwa's Quality of Care Model, which demonstrate specific modifications suggested by culturally diverse women that can potentially improve health care services for these women. PMID- 16340695 TI - An evidence-based project to improve depression and alcohol use screening. AB - Using research to improve practice is a high priority. Research shows that routine screening helps identify adults who are at risk for various disorders. Depression and alcohol use screening tools can improve evaluation and treatment. Nurses aimed to improve the screening rates for depression and alcohol use from the existing 50%-80% to 100% with a 1-hour educational program on depression screening and alcohol use disorders screening for 2 clinic areas: primary care and home-based care. Post program evaluation revealed that depression screening and alcohol use disorders screening rates increased to 100%. PMID- 16340697 TI - Safety of qualitative fit-testing. PMID- 16340698 TI - Environmental tobacco smoke (ets) exposure in Florence hospitality venues before and after the smoking ban in Italy. PMID- 16340701 TI - Lung cancer risk assessment in relation with personal exposure to airborne particles in four French metropolitan areas. AB - OBJECTIVE: The objective of this study was to quantify the total number of lung cancer deaths attributable to chronic exposure to fine particles (pm2.5) among adult populations of four French urban populations (Grenoble, Rouen, Paris, and Strasbourg). METHODS: A geographic information system (GIS) was used to extrapolate personal average exposures to the total population of each metropolitan area across three sectors contrasted in terms of local sources of pollutants. We followed a health impact assessment approach by using the risk estimates in the CSP II study. RESULTS: The annual number of lung cancer cases attributable to pm2.5 chronic exposure ranged from 12 to 404 according to the city. Among these deaths, up to 60% occurred, in the Paris metropolitan area in the urban sector most exposed to traffic emissions. CONCLUSIONS: The health impact of chronic exposures to urban air pollution in metropolitan areas of developed countries warrants further efforts to abate sources of toxicants and to reduce exposure. Traffic emissions still contribute significantly to the total lung cancer burden in France. PMID- 16340703 TI - Lost productive time associated with excess weight in the U.S. workforce. AB - OBJECTIVE: The objective of this study was to examine health-related lost productive time (LPT) in overweight and obese workers. METHODS: Cross-sectional study using data from a national telephone survey of the U.S. workforce. Body mass index defined normal-weight, overweight, and obese workers. LPT in hours and dollars was compared among the three groups. RESULTS: Obese workers (42.3%) were significantly (P<0.0001) more likely to report LPT in the previous 2 weeks than normal-weight (36.4%) or overweight workers (34.7%). Health status mediated the relation between obesity and LPT. Obese workers cost an estimated $42.29 billion in LPT, an excess of $11.70 billion compared with normal-weight workers. Presenteeism accounted for 67.8% of the cost. Comparatively, overweight workers were not a significant source of excess LPT. CONCLUSIONS: Reducing excess weight in the workforce and improving the health of obese workers could positively impact U.S. workforce productivity. PMID- 16340702 TI - Secondary ion mass spectroscopy demonstrates retention of beryllium in chronic beryllium disease granulomas. AB - OBJECTIVE: We hypothesized that beryllium (Be) might persist in lung granulomas in patients with chronic beryllium disease (CBD). METHODS: A total of 33 Be exposed ceramics workers underwent transbronchial biopsy. They were classified based on histopathology and Be-lymphocyte proliferation test as CBD or other categories. Lung tissue sections were analyzed using secondary ion mass spectroscopy. RESULTS: Be was detected in the lungs of all Be-exposed groups. Be levels were increased within the granulomas of patients with CBD compared with the Be levels outside granulomas. Notably, Be was detectable in the lungs of CBD patients who had ceased exposure to Be an average of 9 years previously. CONCLUSIONS: Be was detected in the lungs of all Be-exposed subjects, with the highest levels of persistent Be inside CBD lung granulomas. Be antigen persistence may help explain the chronicity of this granulomatous disorder. PMID- 16340704 TI - Styrene and ischemic heart disease mortality among synthetic rubber industry workers. AB - OBJECTIVE: We examined the relation between styrene and ischemic heart disease (IHD) mortality among 16,579 men in the synthetic rubber industry. METHODS: Associations were measured using stratified and multivariable analysis. RESULTS: Compared with workers with no exposure to styrene, men in the highest quintile of average intensity of exposure (5.50+ parts per million [ppm]) and in the highest quintile of cumulative exposure (60.67+ ppm-years) had IHD rate ratios of 1.14 (95% confidence interval [CI]=0.96-1.35) and 1.07 (95% CI=0.90-1.27), respectively. Acute IHD was not associated with average intensity of exposure within the most recent 2 years or with other indices of exposure. Chronic IHD rates were elevated in subjects with the highest exposure; these associations were weak and imprecise, and evidence of a positive exposure-response relation was limited. CONCLUSION: This study does not indicate that exposure to styrene causes fatal IHD. PMID- 16340705 TI - Sun-protective behaviors of California farm workers. AB - OBJECTIVE: Farm workers are exposed to long hours of solar ultraviolet radiation (UVR), placing them at higher risk for skin cancer. The goal of this study was to evaluate farm workers' sun exposure behaviors. METHODS: Farm workers (n=326) in the San Joaquin Valley of California were interviewed about their sun protective behaviors; these behaviors also were observed directly and these data were used to assess whether participants adequately protected against UVR exposure. RESULTS: All participants were Latino men with a mean age of 33 years. Although rates of wearing any hat and long-sleeved shirts were high, rates of wearing wide brimmed hats and sunscreen were low. CONCLUSIONS: This study is important because there is limited research focused on farm workers' protective behaviors against UVR exposure. Interventions to improve sun protection behaviors of farm workers are needed. PMID- 16340706 TI - Effects of nitrogen dioxide on allergic airway responses in subjects with asthma. AB - OBJECTIVE: We sought to determine whether nitrogen dioxide (NO2) can enhance airway inflammation after allergen challenge in asthmatic subjects. METHODS: Fifteen house-dust-mite (HDM)-sensitive asthmatic subjects were exposed for 3 hours to filtered air or 0.4 ppm NO2, followed by inhalational challenge with HDM allergen. Markers of inflammation were measured in sputum at 6 hours and 26 hours after allergen challenge. RESULTS: After exposure to NO2, eosinophil concentration decreased significantly in the 6-hour postallergen sputum. No significant NO2-related difference was observed for other variables. CONCLUSIONS: Our results suggest that, in most asthmatic individuals, multi-hour exposure to a high ambient concentration of NO2 does not enhance the inflammatory response to subsequent inhaled allergen as assessed by cell distribution in induced sputum. Because the decrease in airway eosinophils has been reported in previous animal studies, future research should be directed toward the mechanism of this effect. PMID- 16340707 TI - Drinking water and dietary sources of nitrate and nitrite and risk of glioma. AB - OBJECTIVE: Dietary nitrite has been associated with increased glioma risk; however, drinking water nitrate has not been extensively evaluated. METHODS: We conducted a population-based case-control study of adult glioma in Nebraska. Water utility nitrate measurements were linked to residential water source histories. We computed average nitrate exposure over a 20-year period. A food frequency questionnaire was used to assess dietary nitrate and nitrite. RESULTS: Increasing quartiles of the average nitrate level in drinking water were not significantly associated with risk (adjusted odd ratios: 1.4, 1.2, 1.3). Risk was similar among those with both higher and lower intakes of vitamin C, an inhibitor of N-nitroso compound formation. Dietary nitrite intake was not associated with risk. CONCLUSIONS: Our study does not support a role for drinking water and dietary sources of nitrate and nitrite in risk of adult glioma. PMID- 16340708 TI - Twenty-four-hour light exposure and melatonin levels among shift workers. AB - OBJECTIVES: We sought to measure melatonin levels and 24-hour light intensity exposure in health care workers over a 7-day period in natural occupational and residential settings. METHODS: Five office workers and 17 nurses working either days or rotating night and day shifts wore a device to record light intensity exposure for one or two 7-day periods, completed a questionnaire, and provided three saliva samples for melatonin. RESULTS: Rotating shift workers had irregular light exposure patterns and abnormal melatonin levels compared with those working days. In addition to lower-than-normal melatonin levels during sleep periods, rotating shift workers exhibited higher-than-normal melatonin levels on arising and during work. Self-reported years of shift work were correlated with measured melatonin and light. CONCLUSIONS: Rotating shift work is supported as a surrogate for exposure to light-at-night and circadian disruption. PMID- 16340709 TI - Upper extremity mononeuropathy among engineers. AB - OBJECTIVES: The objectives of this study were to estimate the prevalence of mononeuropathy at the wrist among engineers who use computers and to identify associated risk factors. METHODS: This is a cross-sectional study of 202 engineers using questionnaires and electrophysiological nerve testing. The definition for median or ulnar mononeuropathy required the combination of distal upper extremity discomfort and abnormal distal motor latency. RESULTS: The prevalence of neuropathy at the wrist among engineers was 10.3% (right median), 3.4% (left median), 1.8% (right ulnar), and 2.9% (left ulnar). Logistic regression analysis identified three variables with positive associations (body mass index, hours of computer use, and antihypertensive medication) and three variables with negative associations (typing speed, driving hours, total break time). CONCLUSIONS: Mononeuropathies at the wrist occur among computer-using engineers and are related to a number of factors, including hours of computer use. PMID- 16340710 TI - Asthma, genes, and air pollution. AB - OBJECTIVE: The objective of this article is to evaluate genetic risks associated with the pulmonary response to air pollutants, including particulates and ozone. METHODS: A comprehensive review of articles related to the genetics of asthma with particular attention to air pollution was conducted through a search of the National Library of Medicine's PubMed database. RESULTS: Asthma, which affects over 15 million people in the United States, is characterized by inflammation leading to reversible airflow obstruction. Triggered by exposure to numerous occupational and environmental agents, asthma has long been considered to occur more frequently in families, with upwards of a 50% higher rate in the offspring of parents with asthma. Asthma genetic studies have used two major methods: mapping techniques that pinpoint gene loci and studies that identify genes and polymorphisms associated with various asthma mechanisms such as inflammatory mediators. The most consistently replicated chromosomal regions associated with asthma have been chromosomes 2q, 5q, 6p, 12 q, and 13q. Because the formation of reactive oxygen species is a major aspect of the inflammatory process of asthma, genetic aberrations associated with antioxidants such as glutathione S transferase (GST) may shed light on reasons why some people with asthma seem more at risk of exacerbations as a result of air pollution. People with a polymorphism at the GSTP 1 locus, which codes for GST, one of a family of pulmonary antioxidants, have higher rates of asthma. Children in Mexico City with the GSTM1 null genotype demonstrated significant ozone-related decrements in lung function. Animal studies support the key role of antioxidants in reducing the inflammatory response associated with exposure to diesel exhaust particles. CONCLUSIONS: Oxidative stress is a key mechanism underlying the toxic effects of exposure to some types of air pollution. Asthmatics with the null genotype for the antioxidant, GST, seem more at risk of the pulmonary effects of air pollution. PMID- 16340711 TI - Incidence of work-related asthma in members of a health maintenance organization. AB - OBJECTIVE: The objective of this study was to evaluate work-related asthma among health maintenance organization (HMO) members. Recent reports suggest that the incidence of work-related asthma may be much higher than Sentinel Event Notification Systems for Occupational Risks (SENSOR) data estimate. METHODS: Using the HMO's electronic medical record, we identified 1,747 persons with evidence of new or recurrent asthma. Interviews with 352 of them elicited information about workplace exposures, symptoms, and home environment. Industrial hygienists rated the potential asthmagenicity of the respondents' work environments. RESULTS: Based on the industrial hygienist ratings and self reported work-relatedness of asthma symptoms, we classified 33% of those interviewed as having potentially work-related asthma, suggesting an overall work related asthma incidence/recurrence rate of 28 cases per 10,000. CONCLUSIONS: The contribution of occupation to the occurrence of adult onset asthma may be much higher than typically suggested in the literature. PMID- 16340712 TI - A randomized clinical trial of continuous low-level heat therapy for acute muscular low back pain in the workplace. AB - OBJECTIVES: We sought to compare the therapeutic benefits of ThermaCare Heat Wrap combined with an education program to an education-only program on reducing pain and disability from acute work-related low back pain. METHODS: Forty-three eligible patients, aged 20 to 62 years who presented to an occupational injury clinic, were randomized into one of two intervention arms: 1) education regarding back therapy and pain management alone or 2) education regarding back therapy and pain management combined with three consecutive days of topical heat therapy (104 degrees F or 40 degrees C for 8 hours). The primary endpoints in this trial were measures of pain intensity and pain relief levels obtained approximately four times per day for the three consecutive working days of treatment, followed by measures of pain intensity and pain relief levels obtained in three follow-up visits at day 4 and 14 from treatment initiation. The secondary measures were overall impairment due to injury and disability caused by low back pain assessed at Intake, Visit 2 (day 4), 3 (day 7), and 4 (day 14). RESULTS AND CONCLUSION: A total of 18 individuals enrolled in the education-only group and 25 in the treatment group completed the intervention and all follow-up visits. The general linear model adjusting for age, sex, baseline pain intensity, and pain medication indicated that the topical heat therapy had significantly reduced pain intensity, increased pain relief, and improved disability scores during and after treatment. PMID- 16340713 TI - Evaluating pulmonary function change over time in the occupational setting. PMID- 16340716 TI - Radio-guided surgery of differentiated thyroid cancer using (131)I or 99mTc Sestamibi. AB - The classical therapeutic approach to patients with differentiated thyroid cancer (DTC) is based on total or near-total thyroidectomy, followed by (131)I treatment and thyroid-stimulating hormone suppressive therapy. This approach allows complete cure in many patients, especially when the tumour is diagnosed at an early stage; it also allows long-term survival in patients with locoregional recurrences or distant metastases if they can be treated with (131)I. In contrast, when metastatic DTC deposits lose their ability to trap (131)I (non functioning metastases), a worse prognosis is expected. Nevertheless, in patients with locoregional non-functioning recurrences, an early diagnosis and prompt surgical extirpation can lead to a favourable prognosis. In these cases, radical surgery is needed. This can be achieved with radio-guided surgery using a hand held gamma probe and a tumour-seeking radiotracer to detect, intraoperatively, the smallest metastatic lesions. In this paper, we discuss the two principal techniques proposed in the literature for radio-guided surgery of non-functioning DTC metastatic recurrences, the first using high doses of (131)I and the second using low doses of 99mTc-Sestamibi. PMID- 16340717 TI - Intra-individual comparison of sentinel lymph node scintigraphy on the day of injection and on the following day in breast cancer. AB - OBJECTIVES: To compare, intra-individually, the detection rates of sentinel node on lymphoscintigraphy performed on the day of injection (D0) and on the following day (D1) in breast carcinoma. We also compared 2-day and 1-day protocols in the two groups of patients. METHODS: The 2-day and 1-day protocols included 76 patients in group 1 and 23 patients in group 2. Patients from group 1 underwent lymphoscintigraphy twice--at 2 h (lymphoscintigraphy 1) and 18 h (lymphoscintigraphy 2) post-injection at four sites periareolar using 99mTc sulfur colloid. Patients from group 2 underwent lymphoscintigraphy only at 2 h post-injection. The detection rates and the number of sentinel nodes were compared in the two lymphoscintigraphy examinations for group 2. RESULTS: The detection rate on lymphoscintigraphy in group 1 was 92% at D0 and 96% at D1. The overall agreement between lymphoscintigraphy 1 and lymphoscintigraphy 2 was 69/76 (91%). In 2/76 women, the sentinel node disappeared at D1 on lymphoscintigraphy, but remained detectable during surgery, and in 5/76 women, the sentinel node appeared at D1 on lymphoscintigraphy. The mean number of sentinel nodes detected on lymphoscintigraphy was 2.05+/-0.14 at D0 and 1.76+/-0.11 at D1 (P=0.004) in group 2, the detection rate of the sentinel node was 20/23 (87%). CONCLUSION: Our study demonstrated that for patients undergoing the 2-day protocol for sentinel node procedure in early stage breast cancer, the optimal imaging time would be to perform lymphoscintigraphy 1 h after injection, with the possibility of imaging patients the following day in cases where lymphoscintigraphy was negative. PMID- 16340718 TI - Can [18F]fluorodeoxyglucose positron emission tomography imaging complement biopsy results from the iliac crest for the detection of bone marrow involvement in patients with malignant lymphoma? AB - OBJECTIVES: To assess the usefulness of [18F]fluorodeoxyglucose positron emission tomography in the detection of bone marrow involvement in malignant lymphoma, and its impact in clinical management. METHODS: One hundred and six consecutive patients with a confirmed diagnosis of lymphoma, referred for staging or restaging of Hodgkin's lymphoma (n=18) or non-Hodgkin's lymphoma (n=88), were reviewed retrospectively. A positron emission tomography scan and bone marrow biopsy of the iliac crest were performed in all patients. The assessment of bone marrow involvement by lymphoma was confirmed by histology and/or progression of bone marrow lesions in clinical follow-up. RESULTS: In 28 of 106 patients, bone marrow involvement was found. Positron emission tomography was more sensitive (86%) than bone marrow biopsy (57%). Positron emission tomography and bone marrow biopsy were concordant by positive correlation in 12 of 28 cases (43%) and by negative correlation in 77 of 78 cases (99%). Ten cases of non-Hodgkin's lymphoma and two cases of Hodgkin's lymphoma with positive positron emission tomography results and an initial negative bone marrow biopsy showed clinical progression of the bone marrow lesions and/or subsequent positive histology. These were considered as false-negative results for bone marrow biopsy. In seven of the 12 positive cases with negative bone marrow biopsy, positron emission tomography uptake distant from the site of the biopsy was seen. In four cases of follicular lymphoma, the bone marrow biopsy was positive and the positron emission tomography scan was normal. CONCLUSIONS: Positron emission tomography and bone marrow biopsy are complementary in assessing the presence of bone marrow involvement in patients with malignant lymphoma. In our series, positron emission tomography was more sensitive than bone marrow biopsy in Hodgkin's and non Hodgkin's lymphoma, except in follicular lymphoma. PMID- 16340719 TI - Impact of FDG PET for staging of Ewing sarcomas and primitive neuroectodermal tumours. AB - AIM: High-grade Ewing sarcomas and Primitive neuroectodermal tumours (PNET) make up the tumours of the Ewing family. Our purpose was to evaluate the value of [18F]fluorodeoxyglucose positron emission tomography (FDG PET) in patients with Ewing tumours. PATIENTS AND METHODS: Twenty-four patients who had PET because of a suspected Ewing tumour during a 5-year period were included in this retrospective study. The images of 33 whole-body FDG PET investigations performed in primary or secondary diagnostics were analysed visually and semi quantitatively by using standardized uptake values (SUVs). In 14 cases, PET was compared to bone scintigraphy regarding bone lesions. The final diagnosis was based on histology, imaging and follow-up. RESULTS: Histologically, the primary lesions were 10 Ewing sarcoma, 13 PNET and one osteomyelitis. The sensitivity and specificity of an examination-based analysis (presence of Ewing tumour and/or its metastases) were 96 and 78%, respectively. Altogether, 163 focal lesions were evaluated. Sensitivity and specificity regarding individual lesions were 73 and 78%. This lower sensitivity is mainly due to small lesions. In true-positive cases, the mean SUV was 4.54+/-2.79, and the SUVs in two false-positive cases were 4.66 and 1.60. True-positive and false-positive cases could not be differentiated definitively based on SUVs because of overlap and low values in true-positive lesions. In four cases, PET depicted 70 while bone scintigraphy depicted only eight bone metastases. CONCLUSION: An FDG PET investigation is a valuable method in the case of Ewing tumours. PET is superior to bone scintigraphy in the detection of bone metastases of Ewing tumours. For the depiction of small lesions, mainly represented by pulmonary metastases, PET is less sensitive than helical computed tomography. Determination of the role of whole-body FDG PET in diagnostic algorithm needs further investigation. PMID- 16340720 TI - 18F-FEAU as a radiotracer for herpes simplex virus thymidine kinase gene expression: in-vitro comparison with other PET tracers. AB - OBJECTIVE: The herpes simplex virus thymidine kinase (HSVtk) gene has frequently been applied as a reporter gene for monitoring transgene expression in animal models. In clinical gene therapy protocols, however, extremely low expression levels of the transferred gene are generally observed. Consequently, sensitive and selective radiotracers for imaging are required. This study describes the in vitro evaluation of 2'-[18F]fluoro-5-ethyl-1-beta-D-arabinofuranosyluracil (18F FEAU) as a candidate tracer for HSVtk imaging with positron emission tomography (PET). METHODS: In cellular accumulation experiments, the potential of 18F-FEAU as a PET tracer for HSVtk was compared to the known acyclic guanosine derivatives 9-[(3-[18F]fluoro-1-hydroxy-2-propoxy)methyl]guanine (18F-FHPG) and 9-[4 [18F]fluoro-3-(hydroxymethyl)butyl]guanine (18F-FHBG), and the thymidine derivatives 3'-deoxy-3'-[18F]fluorothymidine (18F-FLT), 2'-deoxy-2'-[18F]fluoro-5 methyl-1-beta-D-arabinofuranosyluracil (18F-FMAU) and 2'-deoxy-2'-[18F]fluoro-5 iodo-1-beta-D-arabinofuranosyluracil (18F-FIAU). For this purpose, C6 control cells and HSVtk-expressing C6tk cells were incubated with the different tracers for various periods of time and cellular uptake and initial uptake rates were analysed. The initial rate of tracer uptake was determined from the slope of the plot of tracer uptake versus incubation time. RESULTS: After 2 h of tracer incubation, the C6tk/C6 accumulation ratio was 1.6 for 18F-FLT, 2.4 for F-FMAU, 5.5 for 18F-FHPG, 10.3 for 18F-FIAU, 40.8 for 18F-FHBG and 84.5 for 18F-FEAU. The initial tracer uptake rate in C6tk cells was in the order FLT>FMAU>FEAU>FIAU>FHBG>FHPG, whereas the initial tracer uptake rate in C6 control cells was FLT>FMAU>FIAU>FEAU approximately = FHBG approximately = FHPG. The highest HSVtk specific uptake was observed for FEAU. CONCLUSION: This study indicates that the high uptake rate of FEAU together with its high selectivity make this tracer an excellent candidate as a PET tracer for HSVtk gene expression. PMID- 16340721 TI - Discordant localization of 2-[18F]-fluoro-2-deoxy-D-glucose in 6-[18F] fluorodopamine- and [(123)I]-metaiodobenzylguanidine-negative metastatic pheochromocytoma sites. AB - BACKGROUND: Although the majority of pheochromocytomas (PHEO) are benign, a subset is malignant. Computed tomography (CT) and magnetic resonance imaging (MRI) localize PHEO with high sensitivity but, because of limited specificity, [(131)I]- or [(123)I]-metaiodobenzylguanidine ([(131)I]- or [(123)I]-MIBG) is often used as a complementary agent. 6-[18F]-fluorodopamine ([18F]-DA) has been developed as a radiopharmaceutical for the targeting of noradrenergic pathways, and has been shown to result in a better detection rate of PHEO sites than MIBG; however, [18F]-DA has shown a lack of accumulation in some patients with metastatic PHEO. METHODS: Five patients with widespread metastatic PHEO who had CT and MRI evidence of metastatic disease (one man and four women; age range, 25 64 years), and who underwent imaging with [(123)I]-MIBG, [18F]-DA and 2-[18F] fluoro-2-deoxy-D-glucose ([18F]-FDG), were evaluated retrospectively. Tomographic imaging was performed and positron emission tomography (PET) images were inspected visually and quantitatively. RESULTS: All five patients had [(123)I] MIBG scans that grossly underestimated the extent of disease when compared with conventional CT and MRI. All lesions seen on [(123)I]-MIBG scans were detected on [18F]-DA scans, which also detected additional lesions. Nonetheless, [18F]-DA also failed to detect numerous lesions seen on CT and MRI. In all of these cases, [18F]-FDG PET showed lesions that were not detected on either [(123)I]-MIBG or [18F]-DA scans. CONCLUSIONS: When [(123)I]-MIBG or [18F]-DA fails to localize lesions seen on conventional imaging studies, [18F]-FDG may be recommended as an ancillary test for the diagnosis and localization of metastatic PHEO. This is particularly important in patients with aggressive PHEO. PMID- 16340722 TI - A region-of-interest template for three-dimensional stereotactic surface projection images: initial application to the analysis of Alzheimer's disease and mild cognitive impairment. AB - OBJECTIVE: To construct a region-of-interest (ROI) template for Z-score images of three-dimensional stereotactic surface projections (3-D SSP) and to assess whether the ROI template can be a useful tool for evaluation of brain perfusion abnormalities of neurological disorders. MATERIALS AND METHODS: We constructed the ROI template for Z-score images of 3-D SSP based on the standardized magnetic resonance imaging data of 10 healthy volunteers. We assigned a total of 26 ROIs to Z-score images and superimposed it on Z-score images constructed from the brain perfusion SPECT data of 15 patients with Alzheimer's disease and 10 patients with mild cognitive impairment (MCI) who developed Alzheimer's disease within the following 2 years. We then obtained the mean Z-scores of each ROI and examined them to determine whether the hypoperfusion typical of Alzheimer's disease had been demonstrated quantitatively. We also visually inspected the Z score image of each patient in both groups to determine whether the areas with the highest Z-scores were demonstrated within the ROIs of regions typical of Alzheimer's disease. RESULTS: In the patients with Alzheimer's disease, our ROI template quantitatively demonstrated hypoperfusion in regions typical of the disease and the Z-scores were very high. In the MCI patients, the mean Z-scores of the ROI in the posterior cingulated gyrus were the highest among all regions. Visual inspection of the Z-score images of each patient in both groups confirmed that the areas with the highest Z-scores were demonstrated within the ROIs in regions typical of Alzheimer's disease in all cases. CONCLUSION: Use of 3-D SSP methods and our ROI template enables automated quantitative evaluation of brain function images over the entire brain surface. In addition, the ROI template may facilitate visual interpretation of functional images of individual patients with neurological disorders. PMID- 16340723 TI - Simultaneous surface registration of ictal and interictal SPECT and magnetic resonance images for epilepsy studies. AB - BACKGROUND: Subtraction of ictal and interictal single photon emission computed tomography (SPECT) images is known to be successful in localizing the seizure focus in the pre-surgical evaluation of patients with partial epilepsy. A computer-aided methods for producing subtraction ictal SPECT co-registered to the magnetic resonance image (MRI) (the SISCOM method) is commonly used. The two registrations involved in SISCOM are (1) between the ictal-interictal SPECT images, which was shown to be the more critical, and (2) between the ictal image and MRI. OBJECTIVE: To improve the accuracy of ictal-interictal registration in SISCOM by registering all three images (ictal, interictal SPECT, MRI) simultaneously. METHODS: The registration problem is formulated as the minimization of a cost function between three surfaces. Then, to achieve a global minimum of this cost function, the Powell algorithm with randomly distributed initial configurations is used. This technique is tested by a realistic simulation study, a phantom study and a patient study. RESULTS: The results of the simulation study demonstrate that, in surface-based registration, the triple registration method results in a smaller ictal-interictal SPECT registration error than the pair-wise registration method (P<0.05) for a range of values of the cost-function parameter. However, the improved registration error is still larger than that obtained by the normalized mutual information method (P<0.001), which is a voxel-based registration algorithm. The phantom and patient studies reveal no observable difference between registration results. CONCLUSIONS: Although the improved accuracy of triple registration is slightly worse than voxel-based registration, it will soon be possible to apply the results of this study in research utilizing the triple-registration principle to improving voxel based results of ictal-interictal registration. PMID- 16340724 TI - QGS ejection fraction reproducibility in gated SPECT comparing pre-filtered and post-filtered reconstruction. AB - INTRODUCTION: The aim of this investigation was to compare the QGS determined functional parameters using pre-filtering to that using post-filtering in the gated myocardial perfusion single photon emission computed tomography (SPECT) reconstruction process. METHODOLOGY: A total of 25 patient files were examined, each with both a gated rest and gated stress study, and were reconstructed using two strategies. The first employed pre-filtering with a Butterworth low pass filter (order 4.0 and cut-off 0.21) and the second employed post-filtering with a Butterworth low pass filter (order 5.0 and cut-off 0.21). Following reconstruction and reorientation, gated short axis slices were evaluated with QGS software. RESULTS: The mean ejection fraction for the post-filtered data was 49.5% (95% CI, 45.8-53.1%) and for the pre-filtered data was 54.8% (95% CI, 51.4 58.1%). Excellent correlation was demonstrated between the pre- and post-filtered ejection fractions with a correlation coefficient of 0.964. The mean difference between matched pairs of pre- and post-filtered ejection fraction data was 5.3% (95% CI, 4.3-6.3%). The match pair t-test demonstrated a statistically significant difference between matched pairs (P<0.0001) and a statistically significant difference was shown between the means (P=0.005). CONCLUSION: The impact of performing pre-filtering on data in the reconstruction process is significant with a 5.3% increase in the calculated ejection fraction over post filtering. Clearly, this has the potential to undermine diagnostic and prognostic roles of functional parameters. PMID- 16340725 TI - Value of 99mTc-methoxyisobutylisonitrile (99mTc-MIBI) gated SPECT for the detection of silent myocardial ischemia in hemodialysis patients: clinical variables associated with abnormal test results. AB - BACKGROUND: Although coronary artery disease is a major cause of mortality and morbidity in patients undergoing hemodialysis, there is no accurate diagnostic strategy for these patients. AIM: To assess the value of 99mTc methoxyisobutylisonitrile (99mTc-MIBI) gated single-photon emission computed tomography for the detection of silent myocardial ischemia in patients undergoing hemodialysis and to evaluate the clinical variables associated with abnormal test results. METHODS: Thirty-one asymptomatic patients undergoing hemodialysis (20 men, 11 women), with a mean age of 45 years (range, 25-75 years), were included in the study. Serum electrolytes, creatinine, homocysteine and adhesion molecules were measured prior to dialysis. Ambulatory blood pressure, carotid intima-media thickness measurements, echocardiography and stress 99mTc-MIBI imaging were performed in all patients, whereas coronary angiography was performed only in patients with abnormal myocardial perfusion scintigraphy and/or echocardiography. RESULTS: Gated myocardial perfusion scintigraphy results were abnormal in nine patients (29%) and coronary angiography was abnormal in six patients. After a median follow-up of 20 months (range, 14-28 months), nine patients experienced a cardiac event. Seven of the nine patients (78%) with abnormal myocardial perfusion scintigraphy suffered a cardiac event, compared with only two of 22 patients (9%) with normal myocardial perfusion scintigraphy (P<0.0001). Patients with abnormal perfusion scintigraphy had higher serum C-reactive protein, homocysteine and adhesion molecule levels and the duration of hemodialysis was significantly longer. CONCLUSION: In asymptomatic hemodialysis patients, gated myocardial perfusion scintigraphy is a safe and non-invasive screening technique for the detection of coronary artery disease and for predicting future cardiac events. The presence of ischemia correlates significantly with markers of inflammation. The discordant results (abnormal myocardial perfusion scintigraphy/normal coronary angiography) can be attributed to angiographically unrecognized occult atherosclerotic changes and abnormal vasodilatation capacity of the coronary circulation. PMID- 16340726 TI - Characterization of 133Xe gas washout in pulmonary emphysema with dynamic 133Xe SPECT functional images. AB - PURPOSE: To characterize regional ventilation impairment of pulmonary emphysema using dynamic 133Xe single photon emission computed tomography (SPECT) functional images, compared with other forms of chronic obstructive pulmonary disease (COPD). METHODS: Dynamic 133Xe SPECT was performed in 34 patients with emphysema and 15 patients with other forms of COPD. Three-dimensional voxel-based functional images of the half-clearance time (T1/2) mainly reflecting the initial rapid washout of 133Xe gas from the large airways, and of the mean transit time (MTT) reflecting 133Xe gas washout from the entire lungs, including the small airways and alveoli, were created based on an area-over-height method. T1/2 and MTT values were compared with the regional extent of low attenuation areas (%LAA) on density-mask computed tomography images and the diffusing capacity of the lungs for carbon monoxide (DLCO). RESULTS: The MTT/T1/2 ratio in each lung in emphysema was significantly higher than that in other forms of COPD (1.60+/-0.74 vs. 1.21+/-0.26; P<0.01). In the selected unilateral lungs with similar T1/2 values, MTT values were also significantly higher in emphysema. MTT values in each lung showed a significantly closer correlation with the corresponding %LAA values compared with T1/2 values in emphysema (R=0.698, P<0.0001 vs. R=0.338, P<0.01; P<0.05); while only the T1/2 values showed a significant correlation in other forms of COPD (P<0.0001). In correlation with DLCO, MTT values showed a significantly closer correlation compared with T1/2 values in emphysema (R=0.909, P<0.0001 vs. R=0.555, P<0.001; P<0.05); while either value did not show a significant correlation in other forms of COPD. CONCLUSION: MTT values are more critically affected in emphysema compared with other forms of COPD without significant alveolar destruction, and MTT and T1/2 values appear to be differently correlated with the regional extent of LAA between these two disorders. Direct comparison of regional T1/2 and MTT values on functional images may contribute to the demarcation of lung pathology of these two disorders. PMID- 16340727 TI - In-vivo visualization of radiation-induced apoptosis using (125)I-annexin V. AB - BACKGROUND: As apoptosis occurs in tumors within a short time after irradiation, the detection of the frequency of apoptosis may be useful as an indicator of the effect of treatment. For the evaluation of apoptosis under these conditions, tissue extraction from patients is indispensable. AIM: To develop a noninvasive imaging technique to measure and monitor apoptosis in tumor cells caused by X irradiation using (125)I-radiolabeled annexin V. METHODS: The tumors used were human ependymoblastomas, which were transplanted into nude mice. The tumors were irradiated at 2, 5 or 10 Gy. (125)I-annexin V was administered intravenously 6 h after irradiation. In the 5 Gy irradiation group, the isotope was injected at various time intervals (3, 6 and 12 h) after irradiation. Three hours after the injection, the mice were sacrificed, the tumors were quickly removed and frozen sections were prepared at 6 and 40 microm thickness using a cryomicrotome. In autoradiographic imaging, the tumor-to-muscle ratios were compared in the respective irradiated groups. In addition, apoptosis detection by the in-situ end labeling (Klenow) assay was conducted on the same sections. The number of Klenow positive cells was counted in 100 x fields for each section. RESULTS: Both autoradiography and immunohistochemical staining showed a significantly higher frequency of apoptosis in the neoplasms in all irradiated groups than in the control group (P<0.05). Although immunohistochemical staining revealed a peak apoptosis frequency in the 5 Gy irradiated group, autoradiography revealed a peak in the group receiving a lower dose than 5 Gy. When the time from irradiation to annexin injection was varied, both imaging methods showed a peak apoptosis frequency in the group receiving the injection 6 h after irradiation. CONCLUSION: It is possible to predict the effect of treatment in cancer in a noninvasive manner by apoptosis imaging in vivo after radiotherapy. PMID- 16340728 TI - Review of monitoring free muscle flap transfers in reconstructive surgery: role of 99mTc sestamibi scintigraphy. AB - BACKGROUND: Free tissue transfer is a method of moving any tissue from a donor area of the body to a recipient site and re-attaching the arteries and veins to the blood vessels at the recipient site by microvascular surgery. Improved microsurgical techniques have resulted in a high percentage of successful free tissue transfers. Post-operative monitoring of tissue viability can detect early problems in free tissue transfer which may allow early intervention and salvage. Although many flap monitoring methods have been described, there is still no consensus on which of these techniques will become the standard accepted method for monitoring free muscle flaps. OBJECTIVE: In present study, we investigated the use of 99mTc sestamibi scintigraphy in determining free muscle flap viability and complications, and also in directing treatment. METHODS: Thirteen patients were examined prospectively during the post-operative period after free tissue transfer for foot defects. The cause of the defect was diabetic foot ulcer in 10 patients, dermatofibrosarcoma in one patient, squamous cell carcinoma in one patient and gunshot wound in one patient. Foot defect covering was carried out with a free latissimus dorsi muscle flap and skin graft (n=12) and a free gracilis muscle flap (n=1). All patients were examined with a monitoring system that consisted of visual inspection, hand-held Doppler ultrasonography and scintigraphic examinations. Scintigraphic imaging of all cases was performed routinely within the first 48 h post-operatively, and also on days 10 and 91 in two patients. RESULTS: There were four flap failures during the study. One of these patients had viable findings upon visual inspection and no evidence of vascular compromise on Doppler at the first examination. In the other patient, visual inspection of the flap showed that it was ischaemic in one region, but there was no vascular compromise on Doppler examination. Scintigraphic images of each of these patients showed a partial hypoperfused area in the flap region. Later, these two flaps showed positive clinical indications of hypoperfusion (colour of muscle and appearance of skin graft) and Doppler abnormalities. The remaining two patients had non-viable scintigraphic images as well as positive clinical indicators of hypoperfusion and evidence of vascular compromise on Doppler. Nine patients each had a viable flap. In these patients, all three examination tools demonstrated that the flaps were totally viable and there were no vascular complications. CONCLUSION: According to the results of this study, 99mTc sestamibi scintigraphy appears to be a feasible and promising method in the evaluation of free muscle flap viability and complications. On the other hand, to demonstrate any impact on management or patient outcome, further evaluation of 99mTc sestamibi imaging, including comparative studies with different established methods in a larger patient population, is highly recommended. PMID- 16340731 TI - Help your patient downsize with bariatric surgery. PMID- 16340733 TI - Hand hygiene compliance: step up, reach out. PMID- 16340734 TI - Preventing venous thromboembolism. PMID- 16340737 TI - Bone marrow-induced tolerance in the era of pancreas and islets transplantation. AB - Enormous progress has been made in the field of solid organ adaptation recently because of the improvement in immunosuppression. Although powerful immunosuppressive drugs decrease the rate of acute rejection significantly, the long-term functional graft survival and tolerance induction remains poor. Chronic rejection is the main cause of graft failure. An electronic search was performed for articles on chimerism, tolerance, and immunologic perspectives of islet and pancreas transplantation along with referrals to our experience. Infusion of donor bone marrow-derived cells to create a chimeric state continue to be tested in clinical protocols intended to induce specific immunologic tolerance. The proposed mechanisms of immunologic engagement and the emergence of a tolerant state through mixed chimerism include central depletion of alloreactive cells, induction of T-cell anergy, and generation of suppressor cells by interactions between donor and host cells. In this setting, depletion of recipient T cells by different strategies and subsequent repopulation by donor hematopoietic cells after donor bone marrow infusion are prerequisites for tolerance induction. Many efforts have aimed to establish mixed chimerism along with tolerance in solid organ transplantation including pancreas and islets to facilitate engraftment. A review of the more important advances in the field and the future prospects combined with our experience to induce tolerance in the clinic and the laboratory is presented in this article. PMID- 16340738 TI - Persistent normoglycemia in the streptozotocin-diabetic rat by syngenic transplantation of islets isolated from a single donor with Liberase. AB - OBJECTIVES: Enzymatic digestion of donor pancreas is a vital step in islet isolation process. Recently, human and large mammalian islet isolation has been improved by Liberase. However, there are no data to show the improvement of rat islet isolation with Liberase. We hypothesized that commercially available Liberase has variable activities from batch to batch and that a short culture period might improve the function of isolated islets in vivo. We therefore isolated islets with Liberase and cultured them for a short period before transplantation to reverse diabetes in a syngenic rat model. METHODS: Islets of high inbred Lewis rats were isolated with Liberase, purified by discontinuous density gradients, hand-picked, and cultured at 37 degrees C for 24 hours. The same batch of Liberase was used for all experiments in this study. Freshly isolated and cultured islets were implanted intraportally into rats rendered diabetic by streptozotocin. RESULTS: In this study, we were not able to reverse diabetes by syngenic transplantation of freshly isolated islets from 2 donors with Liberase in the rat model. Surprisingly, for the first time, we successfully achieved normoglycemia for more than 100 days in the diabetic rats by syngenic transplantation of the cultured islets isolated with Liberase. These normoglycemic diabetic rats showed normal glucose tolerance curves. Histologic examination of the livers of the islet recipients revealed intact islets, with numerous well-granulated insulin-containing cells and only few glucagon expressing cells. In islets of the recipients' pancreata, the remaining native islets consisted mainly of glucagon-expressing cells, with few insulin-expressing cells in the center. CONCLUSIONS: We conclude that Liberase isolation followed by a short culture period may be a good substitute for collagenase in rats. Islet culture before transplantation can promote the success of single-donor-single recipient islet transplantation to reverse diabetes in the rat model. PMID- 16340739 TI - Autoimmune pancreatitis and complement activation system. AB - OBJECTIVES: Autoimmune pancreatitis is characterized by increased serum level of IgG4, but its pathogenesis has not been fully elucidated. Because this disease is occasionally associated with decreased levels of complements, we sought to clarify which complement activation system was operating in its active state. METHODS: We measured serum levels of complements, mannose-binding lectin, and circulating immune complex in patients with autoimmune pancreatitis, patients with chronic pancreatitis, and healthy controls. RESULTS: We found high serum circulating immune complex values, which decreased significantly after corticosteroid therapy. In patients with autoimmune pancreatitis, elevated levels of circulating immune complex, as determined by C1q assay, were significantly associated with increased serum levels of IgG1 and decreased levels of C4, as well as with a tendency toward decreased levels of C3. There were no significant differences in the serum levels of mannose-binding lectin or in the frequency of a mutant allele of mannose-binding lectin between patients with autoimmune pancreatitis and those with chronic calcifying pancreatitis. Furthermore, corticosteroid therapy had no effect on the level of mannose-binding lectin. CONCLUSIONS: Autoimmune pancreatitis exhibits a high serum circulating immune complex values in its active state, which links to a complement activation system with a classic pathway rather than the mannose-binding lectin pathway or alternative pathways. PMID- 16340740 TI - Decreased total lymphocyte counts in pancreatic cancer: an index of adverse outcome. AB - OBJECTIVES: An impaired host immunity might concur in determining the dismal prognosis of patients with pancreatic cancer (PC). Our aim was to ascertain whether the immunophenotype pattern of blood lymphocytes in PC correlates with tumor stage, grade, or survival. METHODS: We studied 115 patients with PC, 44 with chronic pancreatitis (CP), 23 with tumors of the pancreatico-biliary tract, and 34 healthy controls (CS). Survival data were available for 77 patients with PC. Lymphocyte subsets were determined by fluorescent activated cell sorter (FACS) analysis. RESULTS: In patients with PC, total lymphocyte counts were lower than in CP or CS, and CD8 lymphocyte subset levels were higher with respect to CS. Lower circulating lymphocytes were found in advanced PC stages (IIB-IV; chi2 = 11.55, P < 0.05) compared with stages 0 to IIA. Cox regression analysis, made considering total lymphocyte counts and tumor stage as covariates, was found to be significant for both tumor stage (P < 0.001) and total lymphocyte counts (P < 0.05). CONCLUSIONS: The reduction of total lymphocytes in blood is the main immunologic change in advanced PC. The survival of these patients depends mainly on tumor stage, but it is also affected by the number of circulating lymphocytes, suggesting that the immune system plays an important role in pancreatic adenocarcinoma immunosurveillance and immunoediting. PMID- 16340741 TI - Differences in immune cells engaged in cell-mediated immunity after chemotherapy for far advanced pancreatic cancer. AB - OBJECTIVES: Little is known about the effects of chemotherapy on the immunity of cancer patient or the ideal timing for immunotherapy combined with chemotherapy. To address these questions, we evaluated the effect of gemcitabine and cisplatin combination chemotherapy on the immunity of pancreatic cancer patients. METHODS: Thirteen patients with far advanced pancreatic cancer were enrolled and 7 sex- and age-matched healthy volunteers were included as a control group. RESULTS: Compared with healthy controls, the amounts of peripheral blood mononuclear cells, dendritic cells (DCs), natural killer cells, CD4, and CD8 T cells were reduced. With this numerical suppression, NK cell cytotoxicity to K562 leukemia cells was also significantly impaired (7.7% +/- 4.9% versus 21.7% +/- 7.9% of DNA loss; P = 0.016). Serum concentrations of VEGF and interleukin-10 (IL-10) were higher than the control group (192.1 +/- 114.7 versus 50.8 +/- 39.5 pg/mL of vascular endothelial growth factor (VEGF) and 122 +/- 68.9 versus 111.4 +/- 37.4 pg/mL of IL-10). After 1 cycle of gemcitabine and cisplatin chemotherapy, the impaired immunity of patients with pancreatic cancer was restored. Specifically, the recovery of DCs occurred rapidly and exceeded the value of healthy controls. Levels of the immunosuppressive cytokines, IL-10 and VEGF, gradually decreased during chemotherapy. CONCLUSIONS: Systemic chemotherapy seems to be beneficial for restoring the impaired immunity of patients with pancreatic cancer, and one of the ideal times to collect DCs for immunotherapy is after completing each cycle of chemotherapy. PMID- 16340742 TI - Predicting survival after surgical resection for pancreatic ductal adenocarcinoma. AB - OBJECTIVES: We reviewed the pancreatectomies that were done for pancreatic ductal adenocarcinoma to evaluate patient survival and prognostic predictors. METHODS: A review was performed on 94 patients who underwent surgical resection for pancreatic ductal adenocarcinomas from 1995 to 2002. The perioperative factors were compared between the proximal and distal lesions by the chi2 test and t test. Possible predictors for survival were examined for by univariate and multivariate analysis. RESULTS: The 5-year survival was 16%. The proximal lesions had a smaller tumor size (3.0 +/- 0.11 vs. 3.9 +/- 0.33 cm, respectively; P = 0.03), a higher incidence of nodal involvement (60.6% vs. 34.8%, respectively; P = 0.031), and poorer histologic differentiation (25.4% vs. 13.0%, respectively; P = 0.01) compared with the distal lesions, and both types of lesions had similar rates of intraoperative transfusion, complete resection, and survival. The factors shown to have favorable independent prognostic significance were negative resection margins (hazard ratio [HR] = 0.23; 95% confidence interval [CI] = 0.12 0.42; P < 0.001), a tumor diameter less than 3 cm (HR = 0.46; 95% CI = 0.27-0.78; P = 0.004), well/moderate tumor differentiation (HR = 0.37; 95% CI = 0.19-0.72; P = 0.004), and adjuvant therapy (HR = 0.61; 95% CI = 0.37-0.99; P = 0.49). CONCLUSIONS: For the long-term survival of patients with pancreatic ductal adenocarcinoma, complete excision is the most important therapeutic option, and adjuvant therapy is a significant contributing factor. PMID- 16340743 TI - Gemcitabine and oxaliplatin in patients with pancreatic adenocarcinoma: clinical and pharmacokinetic data. AB - OBJECTIVES: This phase 2 study evaluated clinical efficacy, toxicity, and pharmacokinetics of combination gemcitabine (GEM) and oxaliplatin (OXA) in patients with advanced pancreatic adenocarcinoma. METHODS: Of 30 eligible patients, 20 had metastatic disease and 10 had nonmetastatic unresectable locally advanced disease. Gemcitabine 1000 mg/m2 as a 10 mg/m2/min intravenous infusion on day 1 and oxaliplatin 100 mg/m2 as a 2-hour intravenous infusion on day 2 were administered every 2 weeks. Pharmacokinetics were evaluated in 11 patients by administering the 2 drugs in opposing sequences GEM-OXA (GEM day 1, OXA day 2) and OXA-GEM (OXA day 1, GEM day 2). RESULTS: Of 30 patients evaluated, 9 had a partial response, 11 had disease stabilization, and 10 had disease progression. Median progression-free survival and overall survival were 5.5 and 9.5 months, respectively. The 1-year survival was 37% for all patients. This study revealed no significant pharmacokinetic interaction between the 2 drugs in the GEM-OXA or in the OXA-GEM sequence. CONCLUSIONS: The combination of GEM and OXA was well tolerated and showed a promising activity in patients with advanced pancreatic adenocarcinoma; no sequence-dependent pharmacokinetic interaction occurred when comparing the GEM-OXA versus the OXA-GEM sequence, with a 24-hour interval. PMID- 16340744 TI - A phase II study of weekly cisplatin and gemcitabine in patients with advanced pancreatic cancer: is this a strategy still worth pursuing? AB - OBJECTIVES: A phase 2 study to assess the activity of the cisplatin-gemcitabine combination in patients with advanced pancreatic cancer. METHODS: Chemotherapy naive patients with locally advanced/metastatic/relapsed adenocarcinoma of the pancreas received cisplatin 25 mg/m2 followed by gemcitabine 1000 mg/m2 intravenously on days 1, 8, and 15 of a 28-day cycle. Radiologic response was assessed after 3 cycles, and treatment continued for up to 6 cycles in the absence of disease progression. RESULTS: Thirty-six patients were enrolled, 35 patients were evaluable for toxicity. Hematological toxicity was significant but mostly asymptomatic with grade 3 to 4 (% of patients): leucopenia, 40%; neutropenia, 60%; thrombocytopenia, 60%. There were only 3 episodes of neutropenic sepsis and 2 significant bleeding episodes. Grade 3 to 4 nonhematological toxicities were uncommon but included constipation, infection without neutropenia, lethargy, and thromboembolic events. Of 32 evaluable patients, 62.8% achieved stable disease (SD) or better (SD, 53.4%; partial response, 9.4%). Twenty-nine patients were evaluable for clinical benefit response: 11 (31%) were clinical benefit responders, whereas 13 (36%) remained stable. With complete follow-up, the median time to disease progression was 5.75 months; median survival was 9.5 months, 6-month survival was 72.2%, and 1-year survival was 41.7%. CONCLUSIONS: The combination of gemcitabine and cisplatin is clearly an active regimen and may improve survival based on our 1-year and median survival findings and results from other institutions. However, only an adequately powered randomized controlled trial will assess any real survival benefit over single agent gemcitabine. PMID- 16340745 TI - Effects of depression on the survival of pancreatic adenocarcinoma. AB - OBJECTIVES: Depression frequently predates the diagnosis of pancreatic adenocarcinoma. In other malignancies, depression has been shown to adversely affect survival. The purpose of this study was to assess whether survival after resection for pancreatic cancer is shortened by the pretreatment presence of depression. METHODS: A database of all patients diagnosed with pancreatic cancer was retrospectively reviewed for depression, resection, and chemotherapy and/or radiation therapy. A total of 258 patients were studied; 21% had depression, 19% had surgical resection of the tumor, and 42% were treated with chemotherapy and/or radiation therapy. Survival data was analyzed using Cox proportional hazard regression and life table analysis. RESULTS: The median survival time for all depressed patients with pancreatic cancer was 5 months compared with 4 months for all nondepressed patients with pancreatic cancer (P < 0.9). There was no difference in stage, rate of surgical resection, rate of chemotherapy administration, or rate of radiation therapy use between depressed and nondepressed patients. CONCLUSION: Patients who had undergone surgical resection or chemotherapy and/or radiation therapy had longer survival times than those who did not. Depression, although common among patients with pancreatic cancer, does not affect survival. PMID- 16340746 TI - Distribution of lymphatic vessels in the neural plexuses surrounding the superior mesenteric artery. AB - OBJECTIVES: To investigate whether lymphatic vessels exist in the neural plexuses surrounding the superior mesenteric artery (SMA) and the ultrastructural relationship between neural plexuses and lymphatic vessels. METHODS: A total of 970 serial sections including the structure surrounding the SMA were obtained from 9 cadavers. They were subjected to conventional hematoxylin/eosin staining and immunostaining for the lymphatic marker D2-40. Epithelial membrane antigen and S100 were also immunostained to identify the perineurium and nerve bundles, respectively. RESULTS: Thin-walled, erythrocyte-free vessels staining with lymphatic markers (D2-40) were found in the neural plexuses surrounding the SMA along a full circumference. There seemed to be a distribution correlation between lymphatic vessels and neural plexuses. Lymphatic vessels were not identified within the nerve bundles. The plexuses contained no lymph nodes in any sections. CONCLUSIONS: To our knowledge we report the immunohistochemical visualization of lymphatic vessels in peri-SMA neural plexuses for the first time. Therefore, particular attention should be paid to the lymphatic vessels within neural plexuses as a possible route of invasion and the source of pancreatic cancer recurrence. PMID- 16340747 TI - Conservative management of acute pancreatitis: complications and outcome in a community-based hospital. AB - OBJECTIVES: Acute pancreatitis remains a potentially life-threatening disease. Despite national and international consensus conferences, the management of acute pancreatitis still differs among hospitals and countries. METHODS: This study analyzes the outcome of acute pancreatitis treated by a conservative strategy in a community-based hospital. The strategy followed a conservative interpretation of German consensus guidelines but did not determine medical decisions by a prospective protocol. RESULTS: The analysis included 145 patients treated for acute pancreatitis in the St. Josef Hospital Oberhausen from 1998 to 2002. The etiology was alcoholic in 54.5% and biliary in 26.2% of patients (other causes, 18.6%). Only 1 patient died of septic multiorgan failure (mortality, 1/145; 0.7%). Severe complications occurred in only 22 patients, including renal failure (n = 7), ileus (n = 6), respiratory or cardiac failure (n = 7), need for surgery (n = 4), and sepsis (n = 11). Further complications included pneumonia, delirium, choledocholithiasis/cholecystitis, diabetes mellitus, convulsions, gastric/duodenal ulcers, and pericardial effusion. Of the 145 patients, 75 subjects had at least 1 complication. Fine-needle puncture, mechanical ventilation, hemoperfusion, and hemodialysis were rarely necessary. Computed tomography (CT) was performed in only 69 of 145 patients (47.6%) and showed pancreatic necrosis in 27 of the 69 patients. Maximal C-reactive protein during the first 72 hours and CT findings proved useful in predicting the outcome in univariate analyses. By logistic regression, however, complication rates were associated with Ranson score, but not with CT findings, C-reactive protein, sex, age, etiology, or serum enzymes. CONCLUSIONS: A conservative management of acute pancreatitis in a community-based hospital results in a low rate of complications, mortality, and costs. Clinical assessment (eg, by Ranson score) is sufficient to predict the severity of pancreatitis in most patients. In this cohort, none of the patients were transferred to a specialized center over a 5 year period. Thus, almost all patients with acute pancreatitis can adequately and cost-effectively be cared for in community-based hospitals when following conservative guidelines. PMID- 16340748 TI - Local and systemic effects of hypertonic solution (NaCl 7.5%) in experimental acute pancreatitis. AB - OBJECTIVES: Severe acute pancreatitis (AP) is characterized by hemodynamic alterations and a systemic inflammatory response, leading to a high mortality rate. Treatment of hemorrhagic shock with hypertonic saline solutions significantly reduces mortality through an improvement in the hemodynamic conditions and possibly by an anti-inflammatory effect. Therefore, hypertonic solutions could be effective in AP. METHODS: Wistar rats were divided in 4 groups: group C, control, without AP; group NT, AP, without treatment; group NS, treatment with normal saline solution (NaCl 0.9%) 1 hour after AP; group HTS, treatment with hypertonic saline solution (NaCl 7.5%) 1 hour after AP. AP was induced by injection of 2.5% sodium taurocholate into the pancreatic duct. Mean arterial blood pressure (MAP) and heart rate were recorded at 0 and 2, 4, 24, and 48 hours after AP. After induction of AP, animals were killed at 2, 12, 24, and 48 hours for serum amylase, interleukin (IL)-6, and IL-10 analysis, pancreatic tissue culture and histologic analysis, oxidation and phosphorylation of liver mitochondria, pulmonary myeloperoxidase activity (MPO), and mortality study. RESULTS: In animals of groups NS and NT, a significant decrease of MAP was observed 48 hours after AP (NS: 91 +/- 3 mm Hg; NT: 89 +/- 3 mm Hg) compared with baseline (C: 105 +/- 2 mm Hg) and to HTS group (HTS: 102 +/- 2 mm Hg; P < 0.05). In animals of group NT, NS, and HTS, serum IL-6 and IL-10 levels were significantly higher at 2 hours after AP compared with the control group. However, IL-6 levels at 12 hours after AP and IL-10 levels at 2 and 12 hours after AP were significant lower in group HTS compared with NS and NT groups (P < 0.05). In group HTS, a decrease of pulmonary MPO activity and of pancreatic infection was observed 24 hours after AP compared with NT and NS groups (P < 0.05). A significant reduction on pancreatic acinar necrosis and mitochondrial dysfunction was observed after 48 hours of AP in animals of group HTS compared with groups NT and NS (P < 0.05). A significant reduction on mortality was observed in HTS (0/14) compared with NS (6/17; 35%) and NT (7/20; 35%). CONCLUSIONS: The administration of hypertonic saline solution in experimental AP attenuated hemodynamic alterations, decreased inflammatory cytokines, diminished systemic lesions and pancreatic acinar necrosis, prevented pancreatic infection, and reduced the mortality rate. PMID- 16340749 TI - Applicability of disseminated intravascular coagulation parameters in the assessment of the severity of acute pancreatitis. AB - OBJECTIVES: To evaluate the clinical applicability of the determination of disseminated intravascular coagulation (DIC) parameters in acute pancreatitis. METHODS: The subjects for this study were 139 consecutive patients with acute pancreatitis. DIC parameters were assessed at the initial observation of these patients. RESULTS: The levels of the DIC parameters at admission were significantly associated with the severity and the prognosis of acute pancreatitis. Antithrombin III (AT-III), fibrin/fibrinogen degradation products E, platelet count, D-dimer, and thrombin-AT-III complex at admission showed better area under the receiver operating characteristics curve values compared with C-reactive protein. An AT-III value of 69% at admission was the best cut-off value to predict fatal outcome (sensitivity, 81%; specificity, 86%). CONCLUSIONS: The aggravated coagulation parameters predict a fatal outcome in patients with acute pancreatitis. AT-III level (<69%) was the most accurate marker for poor outcome of acute pancreatitis at admission. PMID- 16340750 TI - Investigation of the mechanisms by which EB1089 abrogates apoptosis induced by 9 cis retinoic acid in pancreatic cancer cells. AB - OBJECTIVES: Previous research has shown that the retinoid 9-cis retinoic acid (RA) promotes apoptosis in pancreatic cancer cells. The vitamin D analog EB1089 does not. Furthermore, cotreatment of cells with 9-cis RA and EB1089 abrogates apoptosis. To explain this, we studied the regulation of proteins involved in apoptotic signaling pathways in pancreatic cancer cells. METHODS: The pancreatic adenocarcinoma cell line T3M4 was used. Cell proliferation was measured using the SRB protein dye assay. Induction of apoptosis was evaluated using an ELISA assay. Caspase activation was detected using a colorimetric assay based on cleavage of a caspase-associated substrate. Regulation of protein levels and posttranslational events were detected using immunoblotting. RESULTS: We confirm that EB1089 diminishes apoptosis induced by 9-cis RA in T3M4 cells. We extend the study to show that EB1089 abrogates increases, induced by 9-cis RA, in caspase activation, p27Kip1 protein levels, Bim and Bax protein levels and in Bax/Bcl2 ratio. In addition, the CDKI p21Waf1 and CAII, a differentiation marker for pancreatic cancer cells are also differentially regulated. CONCLUSIONS: These results suggest that the inhibitory effects of EB1089 on 9-cis RA-induced apoptosis lie upstream of caspase activation and could be associated with reduction of p27Kip1 protein levels. PMID- 16340751 TI - Epidermal growth factor promotes invasiveness of pancreatic cancer cells through NF-kappaB-mediated proteinase productions. AB - OBJECTIVES: Overexpression of epidermal growth factor (EGF) and EGF receptor has been associated with progression and invasive phenotype in pancreatic cancer. However, the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has been poorly understood. In this study, we determined the effect of EGF on the invasiveness and the related regulatory mechanism in two pancreatic cancer cell lines NOR-P1 and KP4. METHODS: Invasion assay was performed to examine the invasiveness of tumor cells induced by EGF, and the expression of matrix metalloproteinase (MMP)-9, MMP-2, and plasminogen activator (uPA) was determined by reverse transcription-polymerase chain reaction and Western blot. Gelatin zymography was used to detect the activities of MMP-9 and MMP-2, and the nuclear factor-kappaB (NF-kappaB) binding activity was determined by electrophoretic mobility shift assay (EMSA). RESULTS: EGF significantly increased the invasiveness of both cell lines but did not affect cell proliferation or adhesion. Increased invasiveness was associated with the induction of MMP-9 and uPA at both mRNA and protein levels. Furthermore, EGF stimulated NF-kappaB binding activity. Moreover, pretreatment of cells with NF-kappaB inhibitors, pyrrolidine dithiocarbamate or ibuprofen, markedly attenuated EGF-induced NF kappaB activation. Subsequently, the EGF-induced MMP-9 and uPA expression and MMP 9 activity, as well as cell invasiveness, were also inhibited by these NF-kappaB inhibitors. CONCLUSIONS: Our findings indicated that NF-kappaB-mediated MMP-9 and uPA induction was responsible for EGF-induced invasiveness in these pancreatic cancer cell lines and implicate that such anti-NF-kappaB therapy as the use of NF kappaB inhibitors may contribute to the reduction of invasiveness of pancreatic cancer. PMID- 16340752 TI - Effects of subtotal colectomy on bacterial translocation during experimental acute pancreatitis. AB - OBJECTIVES: The colon is considered a major source of bacteria causing infection of pancreatic necrosis in acute pancreatitis (AP). Subtotal colectomy before AP in rats reduces mortality, but its role in affecting small bowel flora, bacterial translocation, and infection of pancreatic necrosis is unknown. Our aim was to study these phenomena in rats with AP. METHODS: Fifty rats, allocated in 4 groups, underwent 2 laparotomies: group 1, sham laparotomy and saline biliopancreatic duct infusion; group 2, subtotal colectomy and saline infusion; group 3, sham laparotomy and AP (ductal infusion of glycodeoxycholic acid and intravenous cerulein); group 4, subtotal colectomy and AP. Seventy-two hours later, samples were collected for microbiological analysis. RESULTS: Subtotal colectomy caused small bowel bacterial overgrowth with gram-positive cocci (group 1 versus group 2, duodenum: P = 0.030, ileum: P = 0.029). Bacterial counts of gram-negative rods/anaerobes in the duodenum and ileum and pancreatic bacterial counts of rats with colectomy and AP were significantly higher than in rats with AP only (group 3 versus group 4, duodenum: P = 0.040, ileum: P = 0.029, pancreas: P = 0.017). Duodenal bacterial overgrowth and pancreatic infection correlate significantly (r = 0.45, P = 0.004). CONCLUSIONS: Subtotal colectomy induces small bowel bacterial overgrowth, which is associated with increased bacterial translocation to the pancreas. PMID- 16340753 TI - Histopathologic similarities of inflammatory pseudotumor to autoimmune pancreatitis: a morphologic and immunohistochemical study of 4 cases. PMID- 16340754 TI - A case of intraductal papillary mucinous tumor associated with pancreas divisum. PMID- 16340755 TI - Outcome of out-of-hospital cardiorespiratory arrest in children. AB - OBJECTIVE: To analyze the characteristics and outcome of out-of-hospital cardiorespiratory arrest in children in Spain. METHODS: Secondary analysis of data from a prospective, multicenter study analyzing cardiorespiratory arrest in children. Ninety-five children between 7 days and 16 years with cardiorespiratory arrest. Data were recorded according to the Utstein style. The outcome variables were the sustained return of spontaneous circulation (initial survival), and survival at 1 year (final survival). Neurologic and general performance outcome was assessed by the Pediatric Cerebral Performance Category (PCPC) scale and the Pediatric Overall Performance Category (POPC) scale. RESULTS: Initial survival was 47.3% and 1-year survival was 26.4%. Mortality was higher in children younger than 1 year. Survival of patients with respiratory arrest (82.1%) was significantly higher than survival of cardiac arrest victims (14.4%). Patients who were initially resuscitated by laypersons or paramedics had higher survival (53.6%) than those who were initially resuscitated by doctors and/or nurses (15.2%) (P < 0.01). Mortality was higher in the patients who presented slow rhythms (asystole, severe bradycardia) or pulseless electrical activity than in those presenting ventricular fibrillation (P = 0.001). Multivariate logistic regression revealed that the best indicator of mortality was duration of cardiopulmonary resuscitation longer than 20 minutes. After 1 year, most survivors had normal or mild disability. CONCLUSIONS: Mortality of out-of hospital cardiorespiratory arrest in children is high. When resuscitation is started soon by layperson or paramedics, survival is increased. Duration of resuscitation efforts is the best indicator of mortality. Most of survivors had good long-term neurologic outcome. PMID- 16340756 TI - Unscheduled revisits to a pediatric emergency department: risk factors for children with fever or infection-related complaints. AB - OBJECTIVE: Unscheduled revisits (URVs) may serve as markers of quality of care and may be costly both in financial terms as well as in limitations they place on primary care. We performed this study to examine the association between characteristics easily obtainable during an emergency department (ED) visit and URV to identify a subpopulation of children who may warrant interventions to decrease URV. METHODS: This is a case-control study of patients visiting an urban tertiary care pediatric ED for a fever or infectious disease-related complaint. Cases were defined as patients who had URVs that occurred within 72 hours of an initial ED visit. Control patients were selected by simple random sampling of an enumerated computerized list of all ED visits. Data on independent variables of interest were collected from a chart review and telephone interview with the patient's caregiver. Bivariate and multivariate analyses were performed to determine factors associated with URV. RESULTS: Seventy-five percent of cases and controls participated in the study. Logistic regression analysis revealed 3 factors independently associated with URV for fever or infectious disease-related complaints in children. These included presence of chronic disease (adjusted odds ratio 1.75, 95% confidence interval 1.01-3.03), Medicaid insurance (adjusted odds ratio 1.86, 95% confidence interval 1.04-3.32) and acute triage category (adjusted odds ratio 1.83, 95% confidence interval 1.08-3.10). CONCLUSIONS: These factors may be used to identify children in the ED at greater risk for URV and may point to a need for improved discharge instructions and enhanced communication with primary care and systems to arrange follow-up. Results of this work may also identify at-risk populations for future qualitative research or intervention studies on URV to EDs. PMID- 16340757 TI - Prescription writing errors in the pediatric emergency department. AB - OBJECTIVES: To determine the frequency, type, and severity of written prescription (RX) errors in a pediatric emergency department with attention to specialty and level of training of the residents who wrote the RXs. METHODS: Copies of RXs written by residents in a pediatric emergency department during a 6 month period were matched with pediatric emergency department records. Investigators evaluated individual RXs. Errors were noted and grouped into categories. Severity of errors was scaled, based on predetermined criteria (from previously published articles). The prescribing physician's specialty and level of training were documented. Discharge instructions were reviewed to determine if RX errors were later clarified. RESULTS: There were 358 RXs eligible for overall descriptive analysis. A total of 212 RXs (59%) contained 311 errors. Discharge instructions ameliorated 16% of the RXs containing an error. Minor omissions were the most common error (62%), followed by incomplete directions (23%), dose/directions errors (6%), and unclear quantity to dispense (5%). In analysis of resident specialty and level of training, 339 RXs, written by 47 residents, were included. RXs written by pediatric residents were less likely to have an error (48%) than those by emergency medicine (81%), family medicine (76%), or combined internal medicine/pediatrics residents (100%). First year residents' RXs were errant 59%, second year residents 42%, and third year residents 69% of the time. The majority of RXs with errors (77%) were categorized as insignificant. Nineteen percent of RXs were defined as a problem and were unlikely to be filled. Less than 5% of errors were significant, and none were serious or severe. CONCLUSIONS: RX errors are very common in the pediatric emergency department. Pediatric-specific experience was more influential than level of training on reducing the likelihood of error. PMID- 16340758 TI - Procalcitonin as a diagnostic aid in osteomyelitis and septic arthritis. AB - OBJECTIVES: Plasma procalcitonin (PCT) increases rapidly during bacterial infections but remains low in viral infections and other inflammatory processes. High plasma PCT typically occurs in children with bacterial meningitis, severe bacterial infections, particularly in cases of septic shock or bacteremia, and in renal parenchymal damage. The aim of this study was to test the usefulness of plasma PCT analysis in the diagnosis of osteomyelitis, septic arthritis, and other skeletal inflammatory diseases in pediatric patients admitted because of fever and limping. METHODS: White blood cell count, erythrocyte sedimentation rate, C-reactive protein, and PCT levels were measured in children admitted to the pediatric department with fever, limping, and suspected osteomyelitis or septic arthritis. PCT levels were measured by an immunochromatography assay, based on monoclonal and polyclonal antibodies against katacalcin. RESULTS: Forty four children were evaluated: 12 (27.3%) were diagnosed with osteomyelitis, 11 (25%) had septic arthritis, 5 children (11.4%) were diagnosed as a soft tissue infection, and transient synovitis or reactive arthritis was diagnosed in another 6 children (13.6%). Four children (9.1%) were diagnosed as having juvenile rheumatoid arthritis, and 6 (13.6%) with different diseases. PCT value was elevated in 7 patients (58.3%) with osteomyelitis, and only 3 children (27.2%) with the diagnosis of septic arthritis had a mildly elevated value. Among the children with other diagnosis, there were no positive PCT values (P < 0.001 between skeletal infection and all other diagnosis). CONCLUSIONS: In this study, PCT was found to be a useful marker in the diagnosis of osteomyelitis and not in septic arthritis. A larger group of patients needed to be studied to confirm our findings. PMID- 16340759 TI - Comparison of the epidemiology of human bites evaluated at three US pediatric emergency departments. AB - OBJECTIVES: (1) Using clearly defined methods, provide a current assessment of the epidemiology of human bites sustained by pediatric patients evaluated at a pediatric ED; (2) Compare the frequency, demography, locations, and management of these injuries from the results of the current study to the 2 prior ED studies on pediatric human bites; and (3) Determine if the frequency of these injuries varies by year, gender, or body location. METHODS: The current study involved a retrospective analysis of all visits to an urban, northeastern, United States pediatric ED from 1995 to 2001. Visits for human bites were identified by International Classification of Disease, Ninth Revision, Clinical Modification codes using 2 separate computerized billing databases. Data were extracted from 2 prior published studies for the comparative portion of the study. Frequency estimates and 95% confidence intervals were created using STATA7. RESULTS: For the current study, there were 115 visits for human bites, which comprised 0.04% [0.03-0.05] of total visits to this pediatric ED. Bite visits as a percentage of pediatric ED visits remained the same during the 7 years of the current study. For all 3 pediatric EDs, human bites consisted of much less than 1% of the overall patient volume. The majority (65% [55.8-73.9]) of the patients in the current study were male. When adjusted for the frequency of all patient visits by gender, there was no statistically significant difference in percentage of visits for human bites between males and females (0.05% [0.039-0.063] vs. 0.03% [0.023 0.044]) in the current study. Prior studies did not reveal a statistically significant predominance of males presenting for human bites. As a percentage of visits by age group, human bites were more frequent among adolescents in the current study. Altercations and child play were reported approximately equally as the circumstances contributing to the bites in the current study, although altercations were reported more often in older age groups. In all 3 studies, fewer than 14% of patients were admitted and over 80% presented within 12 or 24 hours after the bite. CONCLUSIONS: Pediatric ED visits for human bites are infrequent and from the current study data, the frequency of visits for these injuries appears to be currently stable. Altercations may not be the main circumstances surrounding all pediatric human bites, yet altercations and human bites might be jointly associated with adolescence. The current study results suggest that most pediatric ED human bite patients are male, although males may not necessarily sustain human bites more frequently than females. Visits for human bite injuries may increase in frequency with age, in comparison to visits for other medical conditions. PMID- 16340760 TI - Sledding-related injuries among children requiring emergency treatment. AB - OBJECTIVES: The aims of the study were to describe sledding-related injuries among children requiring emergency treatment and assess the use of safety measures before and after these injuries. METHODS: This is a study of a consecutive series of patients treated for sledding-related injuries in the emergency department of a large, urban, academic children's hospital during a 5 year period. Follow-up contact with patients' families was made by telephone or mail within 2 months after the injury. RESULTS: One hundred sixty-eight children were treated for sledding-related injuries during the study period. Children ranged in age from 2 to 18 years (mean +/- SD, 9.1 +/- 3.8 years). The most common mechanism of injury was a collision with a stationary object (45.2%). The area of the body most commonly injured was the head/neck (53.6%). Soft tissue injuries were the most common type of injury (43.4%). Children 6 years or younger were more likely to sustain a head/neck injury (P < 0.001; relative risk [RR], 2.60; 95% confidence interval [CI], 1.46-4.64) and lacerations (P < 0.001; RR, 2.45; 95% CI, 1.58-3.82) than older children. Children 7 to 18 years were more likely to sustain an extremity injury (P < 0.005; RR, 1.39; 95% CI, 1.17-1.65), a fracture or dislocation (P < 0.05; RR, 1.31; 95% CI, 1.10-1.55), and a fall off a sled (P < 0.05, RR, 1.30; 95% CI, 1.08-1.55) than younger children. Children 7 to 18 years were also more likely to be admitted to the hospital (P < 0.05, RR, 1.39; 95% CI, 1.20-1.62) than younger children. Helmet use occurred in only 3.0% of the cases. No other safety equipment was used. Sixty-five percent of injuries occurred when adult supervision was present. After the injury, helmet use was reported to be 25.3%, representing a statistically significant increase (P < 0.001). Other safety measures implemented after the injury include change in behavior on the sled (59.5%) and change in location of sledding activity (19.2%). Only 1.0% of parents reported increased adult supervision after the injury event. CONCLUSIONS: More than 50% of sledding-related injuries were to the head/neck region, but only 3% of patients were wearing a helmet at the time of injury. A program to increase helmet use while sledding could have a significant impact on reducing head injuries. Because most injuries occurred when sledders struck a stationary object in their path, checking the sledding location for the presence of obstacles is also an important preventive measure. PMID- 16340761 TI - Critical airway obstruction, superior vena cava syndrome, and spontaneous cardiac arrest in a child with acute leukemia. AB - We report the unusual presentation of a previously healthy girl with sudden cardiopulmonary arrest caused by acute lymphoblastic leukemia and mediastinal involvement leading to acute tracheal and airway obstruction. Despite active resuscitation and mechanical ventilation, she developed severe cerebral edema as a result of cerebral asphyxia. She also had facial edema caused by superior vena cava obstruction, a high peripheral white cell count with blast differentials, and renal failure. Because of severe asphyxia leading to brain death and renal failure, chemotherapy was withheld. Her white cell count spontaneously reverted to reference range without chemotherapy. This report serves to alert clinicians of the oncological emergency of "superior mediastinal syndrome" causing airway and superior vena cava obstruction leading to death in this potentially curable disease. PMID- 16340762 TI - Traumatic spinal epidural hematoma-unusual cause of torticollis in a child. AB - Traumatic spinal epidural hematoma is rare in children. The presentation could be nonspecific, leading to a delay in diagnosis. We present an infant who sustained an epidural hematoma after a "trivial" injury. The delay in diagnosis led to minor neurological deficit at 6-week follow-up. Irritability and torticollis after a neck trauma in an afebrile child should alert to the possibility of spinal cord compression. Early imaging and early decompression will minimize morbidity. PMID- 16340763 TI - Junctional rhythm in association with neuroborreliosis. PMID- 16340764 TI - Esophageal duplication cyst: an unusual cause of respiratory distress in infants. AB - Esophageal duplication cysts are rare congenital anomalies. These should be recognized as a cause of respiratory distress in infants and are usually associated with feeding difficulties. We report 2 such cases of esophageal duplication cyst in infants, causing acute respiratory distress. Excision was curative. PMID- 16340765 TI - Seizure in a child after an acute ingestion of levothyroxine. AB - BACKGROUND: Unintentional ingestion of thyroid hormone preparations is a common occurrence reported to poison control centers. These incidents rarely result in serious outcomes. We report a case of a tonic-clonic seizure occurring in a child after ingestion of a maximum of 3.6 mg of levothyroxine. CASE REPORT: A 3.5-year old boy ingested up to 41 tablets of 88 microg levothyroxine, which was not discovered until the following day. On the third day after the ingestion, he had a single 5- to 10-minute tonic-clonic seizure. Laboratory results were total thyroxine greater than 24.0 microg/dL; free thyroxine, 5.5 ng/dL; and thyroid stimulating hormone, 0.03 mU/L. On the following day, the patient had diarrhea and was hyperactive; heart rate was 144 beats per minute, and mild hyperreflexia was noted on neurological examination. Symptoms were resolved 3 days later. CONCLUSION: This is the second case report of a seizure occurring in a child after an unintentional ingestion of levothyroxine. The maximum amount possibly ingested is much lower than that previously reported. Pediatric ingestion of less than 5 mg of levothyroxine may result in serious outcomes. PMID- 16340766 TI - Seizures in a 20-month-old native of Minnesota: a case of neurocysticercosis. AB - A 20-month-old child, native of Minnesota, was diagnosed with neurocysticercosis. He had no history of international travel or pork consumption. This case and review of the literature emphasize the need to consider neurocysticercosis in any child with nonfebrile seizures in the United States because international travel and exposure to international travelers have become so common throughout the world. PMID- 16340767 TI - Croup-treatment update. PMID- 16340769 TI - Sprained my ankle: now my foot hurts. PMID- 16340770 TI - A Cadillac ride to the emergency department. PMID- 16340771 TI - ECGs in the ED. PMID- 16340775 TI - Brain death and organ damage: the modulating effects of nutrition. PMID- 16340776 TI - Good samaritan kidney donation. PMID- 16340777 TI - Analysis of historical outcomes of composite tissue allograft transplants in nonhuman primates. PMID- 16340778 TI - Diagnosis of hepatic veno-occlusive disease by plasminogen activator inhibitor-1 plasma antigen levels: a prospective analysis in 350 allogeneic hematopoietic stem cell recipients. AB - BACKGROUND: Veno-occlusive disease (VOD) is one of the most serious complications following allogeneic hematopoietic stem cell transplantation (HSCT) and is associated with a high mortality. We conducted a large trial in order to investigate the value of plasminogen activator inhibitor-1 (PAI-1) plasma antigen levels in VOD patients as PAI-1 has been described as a possible diagnostic marker of VOD. METHODS: In all, 350 stem cell recipients were included in our study. PAI-1 levels were analyzed prior to conditioning therapy and then weekly until eight weeks after HSCT. Transplantation-related complications (TRC) including VOD, microangiopathic hemolytic anemia (MAHA), and graft-versus-host disease (GVHD) were recorded weekly throughout the study. RESULTS: Maximum PAI-1 antigen levels were increased in all patients with VOD (n=15; mean 248 ng/ml; 95% CI 183-314 ng/ml). Maximum PAI-1 levels above 120 ng/ml showed a sensitivity of 100% and a specificity of 30.6% for VOD after HSCT. CONCLUSION: Our study underlines that maximum PAI-1 plasma antigen levels not exceeding 120 ng/ml have a strong negative predictive value in the diagnosis of VOD and thus represent a helpful non-invasive tool for exclusion of VOD after HSCT. PMID- 16340779 TI - New-onset gout after kidney transplantation: incidence, risk factors and implications. AB - BACKGROUND: Although cyclosporine use has been associated with an increased risk of new-onset gout after renal transplantation, the incidence and risk factors for new-onset gout have not been reported in the era of modern immunosuppression. METHODS: We conducted a retrospective cohort study of Medicare primary renal transplant patients reported in the United States Renal Data System (USRDS), using Medicare claims data to determine the incidence of new-onset gout. Cox regression analysis was used to calculate adjusted hazard ratios (AHR) for cyclosporine (including separate analysis of Neoral) compared directly with tacrolimus, for the risk of new-onset gout, adjusted for baseline demographic factors and posttransplant renal function. RESULTS: The cumulative incidence of new-onset gout was 7.6% at 3 years posttransplant. The following factors were independently associated with an increased risk of new-onset gout: use of Neoral (vs. tacrolimus, AHR 1.25, 95% CI 1.07-1.47) at discharge, recipient male sex (AHR 1.44, 95% CI 1.25-1.67), older age, higher body mass index, and more recent year of transplant. No other immunosuppressive medications were associated with new-onset gout. Diabetes was associated with a significantly lower risk of new onset gout. The development of new-onset gout was independently associated with decreased patient survival (AHR 1.26, 95% CI 1.08-1.47) as well as death-censored graft survival. CONCLUSIONS: Cyclosporine is an independent risk factor for new onset gout after transplantation. The incidence of new-onset gout appears to be increasing even while the use of cyclosporine is decreasing, and the development of new-onset gout was an independent predictor for death and graft loss in this population. PMID- 16340780 TI - The clinical impact of chronic transplant glomerulopathy in cyclosporine era. AB - BACKGROUND: The clinical impact of chronic transplant glomerulopathy (CTG) on the outcome of kidney allograft receiving calcineurin inhibitors (CNIs) remains uncertain. A retrospective study of renal transplant recipients at Ospedale Maggiore of Milan was undertaken to evaluate the clinical outcome of patients with CTG. METHODS: Among 666 biopsies taken at least 6 months after transplantation (Tx) in 498 transplant patients treated with CNIs, 28 cases (5.6%) of chronic transplant glomerulopathy (CTG) were identified and their clinical features at Tx, at follow-up and graft survival were compared with those of 56 controls transplanted in the same period and with kidney functioning 12 months after Tx. Clinical characteristics at biopsy and at 1 year after Tx were similar in the two groups. RESULTS: After diagnosis graft function deteriorated in 22 patients (78.5%), while it remained stable in 6. Graft loss developed in 92 % of patients with proteinuria >2.5 g/day and in 33 % of those with lower proteinuria (P<0.005). In cases with more severe CTG the rate of graft loss was higher, though not significantly. Graft survival at 10 years was 48% in patients with CTG and 88% in controls (P<0.0001). CONCLUSIONS: The incidence and clinical course of CTG do not seem to be modified by CNI-based immunosuppression. The evolution is unpredictable but the severity of glomerulopathy and proteinuria at follow-up are associated with progression to graft failure. Patients with CTG have a graft survival significantly worse than that of the general population of transplanted patients. PMID- 16340781 TI - In situ dye injection bile leakage test of the graft in living donor liver transplantation. AB - BACKGROUND: Bile leakage after living donor liver transplantation (LDLT) remains a serious problem, resulting in lower survival rates. The aim of this study is to clarify the benefits of in situ leakage testing of the cut surface of grafts in LDLT. METHODS: A total of 135 LDLTs were analyzed. The patients were divided into the following two groups according to the in situ dye injection leakage test of the cut surface: test group (n=40) and control group (n=40). The incidence of bile leakage and the risk factors were identified by analyzing the recipients, donors, and transplantation variables. RESULTS: Bile leakage occurred in 12.5% (10/80) of LDLTs. In the control group, there were nine cases of bile leakage (22.5%). On the other hand, there was only one case (2.5%) of bile leakage in the test group (P<0.05). The bile leakage case in the test group was resolved preservationally. However, 2 of the 9 (22.2%) bile leakage cases in the control group required surgery. CONCLUSION: Although there is biliary complication, especially bile leakage from the cut surface, as an inevitable consequence of LDLT, this study suggests that there is advantage in conducting bile leakage testing to minimize the incidence of bile leakage from the cut surface, which is associated with a high risk of graft failure. PMID- 16340782 TI - Rosiglitazone therapy of posttransplant diabetes mellitus. AB - BACKGROUND: The new onset of posttransplant diabetes mellitus (PTDM) is a common problem after solid organ transplantation. Because insulin resistance plays a significant role in the development of PTDM, we treated 40 consecutive patients with PTDM after liver and kidney transplantation with the insulin sensitizer rosiglitazone (ROSI). METHODS: Thirty-three of 40 patients with PTDM were initially stabilized with twice-daily NPH and regular insulin. All patients subsequently began ROSI 4 mg per day. Patients were followed for a mean of 26 weeks and insulin was adjusted using home blood glucose data and hemoglobin A1C (HBA1C). RESULTS: During 12 months of study, 32/107 liver transplant patients (30%) and 8/205 kidney transplant patients (4%) patients developed PTDM. After 3 4 months of insulin and ROSI therapy, insulin was able to be discontinued in 30/33 (91%) patients with PTDM. In all, 12/40 (30%) patients maintained normal HBA1C levels (5.6+/-0.8%) with ROSI monotherapy, whereas 25/40 (63%) required ROSI and a sulfonylurea to meet this goal. Three of 40 (7.5%) had persistent insulin dependence. 25/40 (63%) continued on 4 mg/day of ROSI, and 15/40 (37%) required an increase to 8 mg/day. Mild edema developed in 13% of patients; significant weight gain did not occur. CONCLUSIONS: Rosiglitazone is the first highly effective oral agent for PTDM. The majority of patients with PTDM may be safely treated with ROSI +/- a sulfonylurea. After the expected 3-6 week delay in the onset of ROSI action, most patients with PTDM will no longer require long term insulin therapy. PMID- 16340783 TI - The significance of a single episode of minimal acute rejection after lung transplantation. AB - BACKGROUND: Bronchiolitis obliterans syndrome (BOS) remains the leading obstacle to better long-term outcomes after lung transplantation. Acute rejection has been identified as the primary risk factor for BOS, but the impact of minimal acute rejection, especially a solitary episode, has usually been discounted as clinically insignificant. METHODS: We performed a retrospective cohort study of 259 adult lung transplant recipients to determine the risk of BOS associated with a single episode of A1 rejection, without recurrence or subsequent progression to a higher grade. The cohort was divided into 3 groups based on the severity of acute rejection (none, single episode of A1, and single episode of A2). We determined the risks of BOS stages 1, 2, 3, and death for each group using univariate and multivariate Cox regression analyses. RESULTS: A solitary episode of A1 rejection was a significant risk factor for BOS stages 1 and 2, but not stage 3 or death, in the univariate analysis. Multivariate Cox regression models confirmed that the risk of BOS attributable to a single episode of A1 rejection was independent of other potential risk factors, such as community acquired respiratory viral infections, number of HLA mismatches, and cytomegalovirus pneumonitis. Likewise, univariate and multivariate analyses demonstrated that a single episode of A2 rejection was a significant risk factor for all stages of BOS but not death. CONCLUSIONS: A single episode of minimal acute rejection without recurrence or subsequent progression to a higher grade is a significant predictor of BOS independent of other risk factors. PMID- 16340784 TI - Different model for end-stage liver disease score block distributions may have a variable ability for outcome prediction. AB - BACKGROUND: The model for end-stage liver disease (MELD) scoring system has become the prevailing criteria for organ allocation in liver transplantation. However, it is not clear if the predictive accuracy of MELD is equally homogeneous in different distribution of MELD score blocks. METHODS: We investigated 472 cirrhotic patients (mean MELD, 14.3+/-5.5), and compared the predictive accuracy of MELD and the corresponding Child-Turcotte-Pugh (CTP) scores in patients with low (<16), intermediate (10-20) and high (>14) MELD score range by using c-statistic for area under the receiver operating characteristic curve (AUC) at different time frames. RESULTS: The MELD scores well correlated with CTP scores at baseline (rho=0.492, P<0.001). Overall, MELD was significantly better than the CTP system to predict the risk of mortality. However, in stratified analysis there were no significant differences between MELD and CTP for the c-statistic in patients with low and intermediate range MELD scores at 3 , 6-, 9-, and 12-month (p values all > 0 1). Among patients with high MELD scores, MELD was consistently more accurate than the CTP system in predicting the mortality at 3- (AUC, 0.715 vs. 0.543, P=0.020), 6- (0.705 vs. 0.536, P=0.003), 9 (0.737 vs. 0.507, P<0.001) and 12-month (0.716 vs. 0.526, P<0.001), respectively. CONCLUSIONS: MELD has a better performance only in a subset of patients with higher MELD scores. The outcome in patients with lower range MELD scores cannot be reliably predicted solely with their MELD scores, and alternative prognostic markers should be used in conjunction to enhance the predictive accuracy. PMID- 16340785 TI - Clinical and genetic risk factors for posttransplant diabetes mellitus in adult renal transplant recipients treated with tacrolimus. AB - BACKGROUND: The present study investigated the incidence of posttransplant diabetes mellitus (PTDM) and calculated the risk of developing PTDM under a tacrolimus-based immunosuppression based on clinical characteristics, tacrolimus pharmacokinetics, and genetic polymorphisms related to tacrolimus pharmacokinetics or diabetes mellitus. METHODS: Seventy nondiabetic adult kidney recipients were studied. Patients with continuous high plasma glucose levels, over 6.5 mg/dl of hemoglobin A1c, or requiring insulin and/or oral antidiabetic agents for more than 3 months after transplantation 6 months postoperatively were diagnosed as having PTDM. Twelve genomic polymorphisms were assessed. RESULTS: Six months after transplantation, 10 recipients (14.3%) developed PTDM. Positive risk factors were age (P=0.019) and body mass index (P=0.038). There were no significant differences in acute rejection rate, total steroid doses, tacrolimus pharmacokinetics or its related to genetic polymorphisms between the two groups. The frequency of PTDM was significantly higher in patients with the vitamin D receptor (VDR) TaqI t allele than in those with the TT genotype (P=0.013). On multivariate analysis, age over 50 years (odds ratio 9.28, P=0.003) and the presence of the VDR TaqI t allele (odds ratio 7.05, P=0.048) were correlated with the development of PTDM. CONCLUSION: The incidence of PTDM was 14.3% in our cohort. Age over 50 years was a risk factor. The presence of the VDR TaqI t allele may also be a risk factor for PTDM, suggesting that genotyping of diabetes related polymorphisms is a possible method of predicting a patient's risk for developing PTDM and would be a valuable asset in selecting appropriate immunosuppressive regimens for individuals. PMID- 16340786 TI - Persistent, asymptomatic, microscopic hematuria in prospective kidney donors. AB - BACKGROUND: Asymptomatic, microscopic hematuria is seen in 8-21% of the general population, has a good prognosis, and is generally not an indication for kidney biopsy. But whether it should preclude kidney donation is unclear. METHODS: Of 512 consecutive prospective donors, 14 (2.7%) continued to have asymptomatic, microscopic hematuria over 1 month. If the medical history, physical examination, and computerized tomographic angiography were unremarkable, and if they still wished to donate, a kidney biopsy was performed. RESULTS: In two prospective donors, hematuria resolved after treatment for urinary tract infection; two others declined donation and were referred to their primary care provider. Kidney biopsy in the remaining 10 showed: two normal; four thin basement membrane nephropathy (TBMN); one nonhomogeneous basement membrane abnormalities; one IgA nephropathy, 5 of 16 glomeruli globally sclerotic; one in a patient with a family history of Schimke's Syndrome, 7 of 30 glomeruli globally sclerotic; and one TBMN and early hypertensive changes without systemic hypertension. Only 4 of the 10 who underwent kidney biopsy donated (two normal, two TBMN). CONCLUSIONS: Kidney abnormalities are common in young, otherwise healthy, prospective kidney donor candidates with persistent, asymptomatic, microscopic hematuria. A kidney biopsy is often abnormal and aids in the decision-making process. PMID- 16340787 TI - High body mass index and short- and long-term renal allograft survival in adults. AB - BACKGROUND: The effect of recipient obesity on kidney allograft survival remains enigmatic. The purpose of this study was to evaluate the effect of donor and recipient body mass index on graft survival. METHODS: Retrospective study of 193 consecutive, adult renal transplants, with at least six months follow-up (mean 24+/-14.1 months). Patients were divided into two groups based upon body mass index (BMI), [weight (kg)/height (m)]: normal (<30.0, n=137) and obese (> or =30.0, n=56). Endpoints were graft loss, defined as either total loss of graft function (return to dialysis) or patient death with a functional graft. Unadjusted and adjusted multivariate analysis techniques, including Kaplan-Meier and Cox proportional hazards regression were used. RESULTS.: Individuals with a BMI > or =30 were not more likely to experience graft loss (O.R. 0.93, 95% C.I. 0.50, 1.72). Rates of acute rejection were not increased in obese recipients. While mortality was not increased in the BMI > 30 group, morbidity, especially surgical, had an increased incidence. The ratio of recipient to donor BMI did not influence graft survival. CONCLUSION: Obese recipients (BMI > or =30.0) were not at increased risk for graft failure. Additionally, matching donor and recipient BMI's would not appear to substantially improve transplant outcome. Obese recipients do have increased posttransplant morbidity and risk all the known health consequences associated with obesity. Careful evaluation and clinical management of obese patients allows for successful kidney transplantation with results equivalent to normal BMI patients. PMID- 16340788 TI - Kidney transplantation into bladder augmentation or urinary diversion: long-term results. AB - BACKGROUND: We report on a single-institutional experience with renal transplantation in patients with severe lower urinary tract dysfunction (LUTD) who underwent bladder augmentation or urinary diversion, and assess the long-term results. METHODS: From September 1987 to January 2005, 255 patients (161 male and 94 female), 7 months to 39 years old of age (median age at time of transplantation 14 years), received 271 kidney transplants. Etiology of end-stage renal disease was LUTD in 83 cases. Among these patients, 24 had undergone bladder augmentation or urinary diversion. RESULTS: We identified two groups of patients surgically treated due to LUTD: group 1 included 16 patients (eight male, eight female) aged 4 to 39 years (median 19 years) with bladder augmentation, whereas in group 2, seven patients (five male, two female) 7 months to 31 years old (median 17 years) with incontinent urinary diversion were reported. In the first group, surgical complications after kidney transplantation included one urinary fistula, one ureteral stenosis. Three patients of second group developed recurrent urinary tract infection. Cumulative graft survival rates of all patients transplanted was 69.4% after 15 years, whereas in the two investigated groups, group 1 and group 2, was 80.7% and 55.5% respectively (P=NS.). CONCLUSIONS: Drainage of transplanted kidneys into an augmented bladder or urinary diversion is an appropriate management strategy when the native bladder is unsuitable. Kidney transplantation in patients with bladder augmentation or urinary diversion for LUTD let achieve similar results to those obtained in the general population with normal lower urinary tracts. PMID- 16340789 TI - Pretransplant pregnancy-associated plasma protein-a as a predictor of chronic allograft nephropathy and posttransplant cardiovascular events. AB - BACKGROUND: Cardiovascular disease and chronic allograft nephropathy (CAN) are two of the main complications observed in patients after renal transplantation. Both appear to be manifestations of the same process, in which inflammation plays a determinate role. Pregnancy-associated plasma protein A (PAPP-A) has been shown to be a marker of acute coronary syndrome and cardiovascular pathology. The objective of this study was to demonstrate whether or not serum concentration of pretransplant PAPP-A is a marker of CAN and a predictor of posttransplant cardiovascular events. METHODS: In all, 178 renal transplants (65% males; 53+/-12 years of age) followed up over the course of 49.3+/-33.6 months were used in this study. During the follow-up period, 19 patients developed CAN (diagnosed by renal biopsy) and 27 patients had a cardiovascular event. Previous to transplantation, the following were determined: ultrasensitive C-reactive protein (CRP) (nephelometry); interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) (immunofluorimetric automatized method), and ultrasensitive PAPP-A (ELISA). RESULTS: A positive correlation was found between PAPP-A and the inflammatory markers (PAPP-A vs. CRP, r=0.218; P=0.004; PAPP-A vs. IL-6, r=0.235; P<0.001; PAPP-A vs. TNF-alpha, r=0.372; P<0.001). The multiple regression analysis showed PAPP-A (relative risk [RR]: 6.4; 95% confidence interval [CI]:1.24-33.11; P=0.027) and CRP (RR: 6.05; 95% CI:1.21-29.74; P=0.028) to be predictors of posttransplant cardiovascular events and PAPP-A (RR: 4.27; 95% CI: 1.03-17.60; P=0.044) and TNF-alpha (RR: 5.6; 95% CI: 1.43-21.83; P=0.013) to be predictors of CAN. CONCLUSIONS: PAPP-A correlated with the inflammatory markers studied (CRP, IL-6 and TNF-alpha). Pretransplant serum concentration of PAPP-A is a predictor of posttransplant cardiovascular events and CAN. PMID- 16340790 TI - Composite tissue allotransplantation: development of a preclinical model in nonhuman primates. AB - BACKGROUND: Composite tissue allotransplantation (CTA) has been recently introduced as a potential treatment for tissue loss secondary to burns, injuries, or resections. However, the optimal strategies to prevent CTA rejection remain undefined. Presently, no CTA model exists to evaluate human-specific immunosuppressants or the relative immunogenicity of all CTA tissues. METHODS: We established a NHP CTA model utilizing a sensate osteomyocutaneous radial forearm flap that avoids functional impairment even in the case of graft loss. The model was evaluated in19 monkeys that underwent auto- or allotransplantation, with or without subtherapeutic immunosuppression to temporarily characterize rejection. RESULTS: Autografts showed no evidence of rejection. Nonimmunosuppressed allografts were rapidly rejected showing a perivenular T-cell infiltrate. This was associated with subsequent alloantibody formation and led to graft thrombosis without prominent dermal infiltration. Subtherapeutically immunosuppressed animals also developed alloantibody and rejected in a delayed fashion exhibiting a marked dermal lymphocytic infiltrate similar in magnitude and distribution to previously reported human cases. CONCLUSION: Our NHP model for CTA is well tolerated by NHPs, results in allosensitization, is responsive to immunosuppression, allows for the evaluation of CTA histology and can be used for the systematic preclinical evaluation of therapeutic maneuvers to improve allograft survival. PMID- 16340791 TI - Role of peroxynitrite anion in renal hypothermic preservation injury. AB - BACKGROUND: Peroxynitrite anions may play a role in normothermic renal ischemia and reperfusion. The purpose of this study was to determine if endogenous peroxynitrite anion is involved in renal preservation injury. METHODS: Experiments were conducted in isolated canine renal tubules and in a canine autotransplant model of hypothermic preservation injury. RESULTS: Isolated renal tubules demonstrated progressive loss of membrane transport function after reperfusion with increasing cold storage times in UW solution as assessed by tetraethylammonium cation transport (TEA). This transport defect was not altered by reperfusion in the presence of WW85, a peroxynitrite decomposition catalyst. Likewise, tubule LDH release was not altered by WW85. Renal tubules did not demonstrate any evidence of peroxynitrite formation after cold storage (0-120 h) or after subsequent reperfusion in vitro as measured by nitrotyrosine adduct formation. Addition of exogenous peroxynitrite (1 mM) directly to freshly isolated renal tubules produced strong nitrotyrosine signals but failed to alter membrane function (TEA uptake). Conversely, SIN-1, a peroxynitrite generator molecule, failed to produce a nitrotyrosine signal in extracted renal tubule proteins but significantly impaired transport function. Finally, function of cold stored canine autografts was not affected by the scavenging of peroxynitrite anions (WW85) before kidney harvest and immediately at reperfusion. Tissue biopsies from cold stored kidney autografts also failed to show evidence of peroxynitrite synthesis either after cold storage (72 h) or after kidney transplantation (60 min reperfusion). CONCLUSIONS: This study concludes that peroxynitrite anions are not formed and are not involved in renal preservation injury. PMID- 16340792 TI - Adenoviral bcl-2 transfer improves survival and early graft function after ischemia and reperfusion in rat liver transplantation. AB - BACKGROUND: Primary graft dysfunction due to ischemia and reperfusion injury represents a major problem in liver transplantation. The related cell stress may induce apoptosis, which can be suppressed by bcl-2. The purpose of the study was to investigate the effect of adenoviral bcl-2 gene transfer on early graft function and survival in rat liver transplantation. METHODS: An adenoviral construct that transfers bcl-2 under the control of a tetracycline inducible promoter was generated (advTetOn bcl-2) and used with a second adenovirus that transfers the repressor protein (advCMV Rep). Forty-eight hours before explantation, donor rats were treated with advTetOn bcl-2/ advCMV Rep (n=7) and doxycyclin, with the control adenoviral construct advCMV GFP (n=8) or with doxycyclin alone (n=8). Liver transplantation was performed following 16 hours of cold storage (UW). Bcl-2 expression and intrahepatic apoptosis was assessed. Bile flow was monitored 90 min posttransplantation. The endpoint for survival was 7 days. RESULTS: Bcl-2 was expressed in hepatocytes and sinusoidal lining cells. This was associated with a significant reduction of apoptotic sinusoidal lining cells and hepatocytes after 24 hours and 7 days. Bile production was significantly higher following bcl-2 pretreatment. Furthermore, bcl-2 transfer resulted in significantly improved survival (100% vs. 50% both control groups). CONCLUSIONS: Adenoviral bcl-2 transfer results in protein expression in hepatocytes and sinusoidal lining cells resulting in early graft function and survival enhancement after prolonged ischemia and reperfusion injury. The inhibition of apoptosis in the context of liver transplantation might be a reasonable approach in the treatment of graft dysfunction. PMID- 16340793 TI - Involvement of CCR5 signaling in macrophage recruitment to porcine islet xenografts. AB - BACKGROUND: Porcine antigen primed and CD4+ T-cell-activated macrophages are capable of both recognition and rejection of porcine xenografts. However, the specific signaling mechanisms involved remains to be addressed. The aim of this study was to examine the role of chemokine receptor and CD40 signaling in macrophage recruitment and graft destruction. METHODS: Macrophages were isolated from rejecting CCR2, CCR5, CD40 and control C57BL/6 mice that were recipients of neonatal porcine pancreatic cell cluster (NPCC) xenografts and were transferred to NPCC recipient NOD-SCID mice. RESULTS: Macrophages isolated from rejecting NPCC xenografts in CD40 and wildtype C57BL/6 mice demonstrated upregulated expression of macrophage activation markers as well as CCR5 and CCR2 genes, and caused pig islet xenograft destruction 8 days after transfer to NOD-SCID recipients. Graft infiltrating macrophages from rejecting CCR2 mice showed a similar activation phenotype and destroyed NPCC xenografts 10 days after transfer to NOD-SCID mice. Blockade of MCP-1 by anti-MCP-1 mAb did not prolong graft survival in CD4+ T cell reconstituted NPCC recipient NOD-SCID mice. By contrast, the graft infiltrating macrophages from rejecting CCR5 recipients showed impaired macrophage activation when compared to control C57BL/6 recipients, and transfer of these macrophages did not result in xenograft destruction in NOD-SCID recipients until day 16 after transfer. Analysis of graft infiltrating macrophages from these rejecting NOD-SCID mice showed an impaired activation phenotype. CONCLUSION: These results demonstrate that CCR5 is involved in both the activation and recruitment of macrophages to rejecting islet xenografts but other pathways are involved. PMID- 16340794 TI - Accumulation of T cells with potent regulatory properties and restricted Vbeta7 TCR rearrangements in tolerated allografts. AB - BACKGROUND: We have previously demonstrated that a short-course treatment with LF15-0195, a 15-deoxyspergualin analogue, induces donor-specific tolerance of cardiac allografts in rats and expansion of splenic CD4CD25 regulatory T cells. METHODS: To further characterize long-term tolerance in this model, we have analyzed the phenotype, regulatory properties and TCR-Vbeta usage of the T cells infiltrating the tolerated allografts. RESULTS: We demonstrate that the tolerated allografts express high levels of FoxP3 transcripts and contain a large number of CD4 T cells, half of which express CD25. Moreover, T cells from these tolerated allografts are very powerful at transferring tolerance to a subsequent allograft recipient, demonstrating the presence of potent regulatory T cells at the site of the graft. Interestingly, the T cells infiltrating the tolerated allografts systematically display restricted Vbeta7 TCR rearrangements. CONCLUSION: These results demonstrate in this model of tolerance, a specific accumulation of T cells with potent regulatory properties and exhibiting restricted Vbeta7-TCR rearrangements at the graft site. PMID- 16340795 TI - IL-4 deficiency prevents eosinophilic rejection and uncovers a role for neutrophils in the rejection of MHC class II disparate skin grafts. AB - BACKGROUND: Acute rejection of MHC class II-disparate bm12 skin grafts by C57BL/6 recipient mice is characterized by massive graft infiltration by eosinophils, together with increased intragraft amounts of IL-4 and IL-5 mRNA. IL-5 blockade prevents the intragraft eosinophil infiltration and prolongs the survival of skin allografts. As the differentiation of T cell precursors into Th2 cells is largely driven by IL-4, we investigated the role of IL-4 in MHC class II-disparate allograft rejection. METHODS: We performed skin grafts from MHC class II incompatible bm12 mice into wild-type C57BL/6 mice (IL-4) or C57BL/6 IL-4 deficient mice (IL-4). Graft survival, in vitro T cell reactivity, and histology were compared. RESULTS: We observed that 50% of IL-4 mice rapidly rejected their bm12 allograft, whereas the other 50% retained their graft 60 days after transplantation. Histological examination of bm12 allografts retained by IL-4 mice showed a normal appearance with no inflammatory infiltrate and no eosinophils. Among IL-4 mice that acutely rejected their bm12 skin graft, we observed a dense polymorphonuclear infiltrate. The depletion of neutrophils significantly prolonged bm12 graft survival. CONCLUSIONS: Eosinophil infiltrates, typical of MHC class II disparate acute skin graft rejection, are critically dependent on the availability of IL-4. IL-4 mice reject MHC class II disparate skin grafts by a pathway of rejection where neutrophils play a direct causal role. PMID- 16340796 TI - alpha1,3-Galactosyltransferase gene-knockout pig heart transplantation in baboons with survival approaching 6 months. AB - BACKGROUND: The recent generation of alpha1,3-galactosyltransferase gene-knockout (GalT-KO) pigs has allowed investigation of the survival of GalT-KO pig organs in nonhuman primates. METHODS: Heterotopic heart transplantation from GalT-KO pigs was carried out in baboons (n=8) using a human antihuman CD154 monoclonal antibody-based immunosuppressive regimen. RESULTS: In six of the eight cases, graft survival extended to between approximately 2 and 6 months. All grafts developed thrombotic microangiopathy (TM). In particular, the clinical course of one baboon in which the graft functioned for 179 days is summarized. This baboon received aspirin (40 mg on alternate days) from day 4 in addition to heparin, which may have been a factor in the delay of onset and progression of TM and in prolonged graft survival. Maintenance therapy with anti-CD154 mAb, mycophenolate mofetil, and methylprednisolone was associated with persistently low numbers of CD3CD4 and CD3CD8 cells. Despite persisting depletion of these cells, no infectious complications occurred. CONCLUSIONS: It remains to be established whether TM is related to a very low level of natural preformed or T-cell-induced antibody deposition on the graft, inducing endothelial activation and injury, or to molecular incompatibilities in the coagulation mechanisms between pig and baboon, or to both. However, function of a pig organ in a baboon for a period approaching six months, which has not been reported previously, lends encouragement that the barriers to xenotransplantation will eventually be overcome. PMID- 16340797 TI - Effects of long-term administration of high-dose recombinant human antithrombin in immunosuppressed primate recipients of porcine xenografts. AB - BACKGROUND: Fibrin deposition is central to the acute humoral rejection process occurring in the presence of consumptive coagulopathy when pig organs are transplanted into primates. METHODS: To assess whether strategies aimed at preventing fibrin formation may extend xenograft survival, we administered high daily doses of recombinant human antithrombin (rhAT) (500 U/kg twice daily) to obtain both anticoagulant and anti-inflammatory effects in immunosuppressed primate recipients of porcine kidneys. RESULTS: Some degree of consumptive coagulopathy developed in both rhAT-treated (n=3) and untreated (n=3) primates. No major differences in the coagulation parameters analyzed were observed between the 2 groups. Similarly, no difference in survival was seen between rhAT-treated (20.6+/-4 days; range: 15-23 days) and untreated animals (17.3+/-11.6 days; range: 7-30 days), although the rhAT-treated primates had a higher bleeding tendency. Despite the high daily dose of rhAT, considerable fibrin deposition was observed in the graft as early as 2 weeks after transplantation. CONCLUSIONS: These results suggest that a high daily dose of rhAT fails to influence survival or prevent fibrin formation and deposition in the graft in our pig-to-primate model. However, the potential role of rhAT administered in combination with heparins or other clotting inhibitor concentrates in this model remains to be determined. PMID- 16340798 TI - Detection of antibodies to HLA-DP in renal transplant recipients using single antigen beads. AB - With the development of single DP-antigen beads, antibodies to DP could clearly be segregated from other HLA antibodies. We studied 323 sera of patients from four different centers with functioning or rejected kidney grafts. DP antibodies were found in 5.1% of 138 patients with functioning grafts, and 19.5 % of 185 patients with rejected grafts (P<0.001)). 42.9% and 63.9% of the DP antibody positive patients had DR/DQ antibodies among the functioning and rejected group. Among patients who did not have Class I and DR/DQ antibodies, 13% of those who rejected a graft had DP antibodies, compared to 3.5% of patients with functioning grafts (P<0.05). DP 0301 had the highest specificity frequency in the rejected group. In conclusion, HLA-DP antibodies were detected at a higher frequency in patients who have rejected their grafts than those with functioning grafts. For regrafts, DP tissue typing is recommended to avoid presensitized DP alleles. PMID- 16340799 TI - Long-term kinetic of T-lymphocyte subsets in kidney-transplant recipients: influence of anti-T-cell antibodies and association with posttransplant malignancies. AB - A low CD4 lymphocyte count has been associated with skin and nonskin cancers in kidney transplant recipients (KTR) and might be a useful prediction tool. We analyzed the serial measurements of CD4 and CD8 lymphocytes to study the influence of immunosuppressants and look for a possible predictive value of lymphocyte count for the occurrence of cancer. In 250 successive KTR, blood lymphocytes were labeled for CD4 and CD8 count, prior transplantation, at month 3 and 6 and each year thereafter, throughout a 10-year observation period. The kinetics of lymphocyte subsets were compared in regard to treatments, cancer and confounding factors. ATG were responsible of an early and long lasting impairment in CD4 cells. Patients with a cancer and especially those with multiple tumors, presented with less CD4 cells throughout the observation period, but ATG were not directly associated to the occurrence of tumors. An early low CD4 count was of poor predictive value for the occurrence of cancer. PMID- 16340801 TI - Simplified rat model of intestinal transplantation. PMID- 16340800 TI - Prolonged allogeneic marrow engraftment following nonmyeloablative conditioning using 100 cGy total body irradiation and pentostatin before and pharmacological immunosuppression after transplantation. AB - In a canine model of dog leukocyte antigen (DLA)-identical nonmyeloablative marrow transplantation including postgrafting immunosuppression with cyclosporine (CSP) and mycophenolate mofetil (MMF), engraftment was only transient with 100 cGy total body irradiation (TBI) conditioning, indicating suboptimal pretransplant immunosuppression. In contrast, grafts after 200 cGy TBI were durable in 11/12 recipients. We hypothesized that addition of pentostatin before transplantation could, in part, substitute for 100 cGy TBI. Pharmacokinetic studies showed pentostatin significantly inhibited adenosine deaminase in canine lymphocytes. Eight dogs were conditioned with 6x4 mg/m pentostatin and 100 cGy TBI, whereas two dogs received 3x4 mg/m pentostatin plus 100 cGy TBI. All were given MMF/CSP posttransplant. All showed initial engraftment; four remained stable mixed chimeras for >32 weeks. The median duration of engraftment was 13 (range 9 to >39) weeks, which was significantly longer than in six historical controls conditioned with 100 cGy TBI alone (median 10, range 3-12 weeks) (P=0.01). PMID- 16340805 TI - [Guidelines for the rehabilitation of chronic obstructive pulmonary disease. French Language Society of Pneumology]. PMID- 16340802 TI - Gamma delta T cells and resolution of cytomegalovirus infection in an HIV/HCV coinfected patient after liver transplantation. PMID- 16340806 TI - [Question 1. Respiratory therapy for chronic obstructive pulmonary disease]. PMID- 16340807 TI - [Question 2. Indications for respiratory therapy for chronic obstructive pulmonary disease]. PMID- 16340808 TI - [title]. PMID- 16340809 TI - [Question 3-1. Evaluation of chronic obstructive pulmonary disease at rest]. PMID- 16340810 TI - [Question 3-2. Functional evaluation of respiratory muscles]. PMID- 16340811 TI - [Question 3-3. Evaluation of aerobic capacity]. PMID- 16340812 TI - [Question 3-4. Physiopathology of dyspnea]. PMID- 16340813 TI - [Question 3-5. Evaluation of dyspnea of effort]. PMID- 16340814 TI - [Question 3-6. Physical activity questionnaires - application to chronic obstructive pulmonary disease]. PMID- 16340815 TI - [Question 3-7. Six minute walking test in respiratory therapy]. PMID- 16340817 TI - [title]. PMID- 16340816 TI - [Question 3-8. The shuttle walk test]. PMID- 16340818 TI - [Question 4-1. Physical rehabilitation in the patient with chronic obstructive pulmonary disease]. PMID- 16340819 TI - [Question 4-2. Supplemental oxygen therapy and rehabilitation of the patient with chronic obstructive pulmonary disease]. PMID- 16340820 TI - [Question 4-3. The role of non-invasive measurement of ventilation during exercise, electrostimulation of the quadriceps, and the work of breathing in the rehabilitation of patients with chronic obstructive pulmonary disease]. PMID- 16340821 TI - [Question 4-4. Patient education in chronic obstructive pulmonary diseases]. PMID- 16340822 TI - [Question 4-5. Assessment and management of psychological issues in patients with chronic obstructive pulmonary diseases]. PMID- 16340823 TI - [Question 4-6. Smoking cessation and pulmonary rehabilitation]. PMID- 16340824 TI - [Question 4-7. Activities of daily living in patients with chronic obstructive pulmonary disease]. PMID- 16340825 TI - [Question 4-8. Occupational therapy in the patient with chronic obstructive pulmonary disease]. PMID- 16340826 TI - [Question 4-9. Nutritional management in chronic obstructive pulmonary disease: the role of anabolic agents]. PMID- 16340827 TI - [Question 5. Strategies in respiratory therapy]. PMID- 16340828 TI - [Question 6. Survival analysis and long-term evaluation of the efficacy of respiratory therapy for chronic obstructive pulmonary disease]. PMID- 16340829 TI - [A special issue of the Revue des Maladies Respiratoires dedicated to the courses of the Groupe d'Oncologie de Langue Francaise, in 2005]. PMID- 16340830 TI - [Radiological screening for lung cancer]. AB - Screening for lung cancer using thoracic CT scanning is a topical question. Pilot studies in Japan, the United States and France have demonstrated the high sensitivity of CT scanning for identifying pulmonary nodules. By contrast the specificity of the technique is poor requiring either repeat scanning after an interval or further investigations. Randomised studies will reveal if CT screening can have an impact on mortality from lung cancer. PMID- 16340831 TI - [Assisting smoking cessation]. AB - Smoking is the most important cause of preventable death in the world and the benefits of smoking cessation are clearly established regardless of the age of the smoker or the presence or absence of pathologies, whether caused by smoking or not. Even though this may appear obvious it is useful to recall that every medical consultation for any condition should include an assessment of smoking status, of the motivation to quit (which should be identified and reinforced) and of the readiness to quit. Assistance given can thus be adapted appropriately to the individual situation and should take into account validated recommendations for best practice. Advising and assisting smoking cessation requires a tailored approach which involves a number of elements centred on the addictive nature of smoking. The nature of dependence may differ and be of varying intensity in different individuals. The capacity to give up may also be very different. Obstructions to smoking cessation are often numerous and complex. Co morbidities and co-addictions can slow down the dynamics of change and their existence must be incorporated into an individualised approach. The transition of a smoker towards abstinence passes through numerous steps so that the addicted smoker is effectively a patient with a chronic disease. This explains many relapses and justifies the need to offer often specialised assistance over a prolonged period. Advising and assisting smokers to quit is a priority. It is thus necessary to organise the management of this process according to the specific skills of each health professional. This approach will address the global problem of tobacco while focusing on the specific problems of each smoker. PMID- 16340832 TI - [Epidemiology of tobacco smoking]. AB - INTRODUCTION: The aim of this review is to give an overview on tobacco consumption in France and to describe the mortality and the morbidity related to smoking. STATE OF ART: Following the increase of tobacco prices in 2002-2003, the prevalence of smoking was estimated to 30% in 2004 in France. Tobacco smoking leads to 66,000 deaths annually in France: 59,000 among men and 7 000 among women. Three groups of diseases are linked with smoking: cancers, cardio-vascular diseases and chronic lung diseases. A relation was also established with gravidic or osteoporotic complications. Passive smoking is also involved in numerous diseases such as lung cancer, coronary diseases and strokes. PERSPECTIVES: Smoking cessation conducts to a progressive decrease in morbidity risk. A functional improvement is obtained for patients suffering from chronic lung diseases. CONCLUSIONS: Effective smoking cessation treatments are available and should be systematically offered to smokers. PMID- 16340833 TI - [Thrombosis and lung cancer]. AB - Venous thromboembolism (TE) is a frequent occurrence in lung cancer patients and can be precipitated by venous stasis, the activation of clotting cascades by pro coagulant tumour factors and by vascular injury due to chemotherapy and central venous catheters. Subcutaneous low molecular weight heparin (LMWH) has replaced intravenous unfractionated heparin in the treatment of acute TE. The use of long term LMWH is recommended to prevent the recurrence of TE. Prophylactic treatment is indicated in patients undergoing surgery or hospitalised. Heparin appears to have distinct anti-neoplastic as well as anticoagulant effects, which benefit overall survival. PMID- 16340834 TI - [Bronchoscopic early detection of lung cancer]. AB - The centrally located lung cancer mainly corresponds to squamous cell carcinoma localized in the large airways. In this paper, tools that are used for the detection of this cancer at the earliest stages are reviewed. Fluorescence bronchoscopy is the latest major advance for the early detection of this cancer. PMID- 16340835 TI - [PET scanning in the initial assessment of lung cancer]. AB - 18FDG-PET scanning enables the imaging of metabolic activity giving an assessment of the local extent of thoracic malignancies as well as an indication of the presence of nodal or metastatic spread. This enables more accurate staging and has revolutionised the management of lung cancer. National and international guidelines describe the role of this technique. PMID- 16340836 TI - [Mesothelioma: advances in chemotherapy]. AB - Chemotherapy is often the only treatment possible for locally advanced or metastatic mesothelioma. This paper recalls which drugs might have therapeutic benefits in this condition and reviews recent studies of chemotherapy or targeted therapy. If the patient cannot be enrolled in a therapeutic trial the first line therapy in the absence of contraindications is a combination of cisplatin and pemetrexed, the latter having received a licence for this indication in September 2004. Among the alternatives reviewed are taxanes, liposomal anthracyclines, topoisomerase inhibitors, cisplatin derivatives, vinca alkaloids, and antimetabolites. Although the first three have show little or no benefit the vinca alkaloids (vinorelbine, vinflunine) and particularly the antimetabolites (gemcitabine, raltitrexed, pemetrexed) are very promising. Recent studies have looked most frequently at combinations of an anti-metabolite and a platinum salt, with data available from nearly 200 patients treated with gemcitabine. These studies have had fairly homogeneous results showing a one year survival of about 50%. Some preliminary data from studies of second line chemotherapy is also available. Finally studies of targeted therapies such as anti-EGFR, anti VEGF and anti PDGF are underway but have not as yet demonstrated major therapeutic benefit. PMID- 16340837 TI - [Lung cancer in women]. AB - BACKGROUND: Since the end of the 1980's, there has been a significant increase in the incidence of lung cancer in women in France. This is largely due to an increase in women smoking since the end of the nineteen-sixties, some 20 years later than occurred in North America. STATE OF THE ART: Since 1995 lung cancer has been the third most common malignant cause of death in French women having become the most common in The United States in 1987. The epidemiology of the disease in women is different, with tobacco smoking explaining only 70% of the incidence. Moreover, even if smoking is the primary cause of lung cancer, the reported duration and amount smoked is generally speaking lower than in men with equivalent disease. Other risk factors such as higher expression of the gastrin releasing peptide (GRP) receptor, hormonal factors, radon and passive smoking may also play a role. The distribution of histological types is also different in women, with a predominance of adenocarcinomas. Finally, prognosis appears to be better in women. PERSPECTIVES: Targeted therapies have introduced another gender distinction since women are more likely to respond to gefitinib and erlotinib than men. CONCLUSION: The massive increase in lung cancer in women is the most important epidemiological feature of recent years. Due to some differences in risk factors, histology distribution, prognosis and therapeutic response, specific studies devoted to female patients especially non-smokers are needed. PMID- 16340838 TI - [Lung cancer in elderly patients]. AB - Lung cancer occurring in elderly patients is an increasingly frequent clinical problem both because of an aging population and the general increase in incidence of the disease. The management of lung cancer in the elderly represents therefore a significant public health problem. The full range of therapeutic modalities may be used in these patients provided a comprehensive geriatric assessment is made, not limited to performance status assessment, but also taking into account comorbidities, which are frequent in elderly people, and functional capacities. Elderly patients are often managed in a sub-optimal fashion. With appropriate, equal treatment their prognosis is equivalent to that of younger patients. PMID- 16340839 TI - [Bronchioloalveolar carcinoma]. AB - Bronchioloalveolar carcinoma is one of the four histological subtypes of adenocarcinoma and its incidence is increasing. It grows in a lepidic fashion along the alveolar septa without invading alveolar walls. The strict histological definition requires complete surgical resection to exclude any evidence of invasion. This definition is thus only applicable in practice to isolated operable pulmonary nodules. For other types high-resolution lung CT-scan is necessary to evaluate pulmonary involvement because of the high frequency of multifocal disease at initial presentation and because of the presence of ground glass opacities, which can be one of the first manifestations of CBA on CT. Therapeutic management does not differ from non-small cell lung cancer. Solitary nodules are treated by surgical resection with a good prognosis whereas multifocal disease is rarely respectable. Diffuse and/or pneumonic forms are treated with systemic chemotherapy with a generally poor prognosis. However, the recent discovery of the particular sensitivity of this form of adenocarcinoma to EGFR (Epidermal Growth Factor Receptor) tyrosine kinase inhibitors offers new possibilities for management. PMID- 16340840 TI - [Prognostic factors in lung cancer]. AB - INTRODUCTION: Knowledge of prognostic factors is important for therapeutic decision making but above all, at a population level, for the interpretation of data in the literature and as a guide to clinical research. To be useful in practice they must be both independent and reproducible. Despite an impressive number of publications few descriptions achieve this standard in lung cancer. BACKGROUND: The TNM classification plays a predominant role in the choice of treatment, in prognosis and as an index of performance. Depending on the population studied (small cell or non-small cell cancer, operable or not) other independent factors, identified in large series, improve the prediction of prognosis: sex, age, loss of weight, LDH, leukocytosis, neutrophilia, haemoglobin, serum calcium, NSE, Cyfra 21-1. VIEWPOINT: The numerous biological factors described should be the object of further investigations but may lead to the assessment of new therapeutic strategies. CONCLUSION: Large multicentre studies are essential before a role may be attributed to an independent prognostic factor and before it may be used as a guide to the management of a patient or to improve our prediction of the progress of a patient or a group of patients. PMID- 16340841 TI - [Treatment of thoracic malignancy--results of meta-analyses]. AB - Meta-analyses dealing with the treatment of thoracic malignancy (non-small cell lung cancer, small cell lung cancer and mesothelioma) were reviewed including isolated ones, based on individual data from clinical trials, or conducted in the context of a systematic review of the literature Their results have changed clinical practice, notably by validating radiochemotherapy for patients with unresectable limited diseases, adjuvant chemotherapy for resectable non-small cell lung cancer and by identifying the most active drugs. However it is important to understand the limits of their methodology in order to avoid inappropriate interpretations. PMID- 16340842 TI - [Postoperative follow-up in non-small cell lung cancer]. AB - INTRODUCTION: Survival of patients after surgery for non-small cell lung cancer is significantly limited because of frequent fatal recurrences of the disease. Logically, follow-up should detect recurrences early, thus increasing chances of cure. STATE OF THE ART: Only non-randomised studies have been published. These suggest that thoracic recurrences are the most frequent isolated recurrences and the most frequently treated with curative intent; and that diagnosis of recurrences while patients are still asymptomatic might improve survival. Several guidelines have been published, with follow-up programs of varying intensity and with a recent tendency to reduce follow-up procedures to clinical assessment only (American Society of Clinical Oncology 2004). PERSPECTIVES: All guidelines agree that there is a need for randomised data. Only one randomised trial is ongoing, conducted by the Intergroupe Francophone de Cancerologie Thoracique (IFCT). This study compares a minimal follow-up with physical examination and chest X-ray alone to a more intensive follow-up program reflecting routine French practice which in addition includes thoracic CT scan and fibre optic bronchoscopy. CONCLUSIONS: As it is not yet possible to define the optimal follow-up after surgery for non-small cell lung cancer from existing data, the IFCT randomised study represents for pulmonologists, oncologists and thoracic surgeons a good opportunity to rationalise postoperative follow-up and to defend their practice from minimalist recommendations. PMID- 16340843 TI - [Pain management in bone metastasis of pulmonary origin: new interventional and metabolic techniques]. AB - Invasion of bone by a metastatic lesion is the most common cause of pain in cancer patients. Pain management in these patients is an important and difficult task. The pain is not always properly controlled by high doses of specific medication, radiation therapy or chemotherapy. When these therapies do not provide adequate pain relief, percutaneous vertebroplasty, cementoplasty, radiofrequency ablation and internal radiotherapy appear to be elegant and efficient complementary alternative pain control methods. PMID- 16340844 TI - [Lymph node clearance]. AB - The goal of mediastinal sampling or a radical lymph node resection after lung cancer resection is to correctly stage the illness to enable a more accurate assessment of prognosis. Metastasis to mediastinal nodes may be encountered even in small peripheral T1 tumours. Radical lymph node resection often uncovers metastases that would have been missed by mediastinal sampling alone. The morbidity associated with radical node resection is very low, and long term survival appears to be improved, compared with node sampling. At present, lymph node staging has little influence on the choice of adjuvant treatments. PMID- 16340845 TI - [Interventional bronchoscopy in the management of lung cancer]. AB - The techniques of interventional bronchoscopy have acquired an important place in the management of lung cancer. Techniques to relieve obstruction requiring the use of rigid bronchoscopy under general anaesthetic were the first to be introduced. Laser therapy, which was initially used for coagulation can be replaced by electrocautery. The development of endobronchial stents provides a natural complement to techniques aimed at clearing bronchial obstruction by helping to maintain patency in the long term. Other therapeutic techniques can be deployed in association with those already mentioned, including photodynamic therapy and brachytherapy using low doses of radiation applied locally. These have particular potential for the treatment of early disease. Questions remain as to the precise indications for the different techniques and of how to identify which approach is most appropriate for individual patients. PMID- 16340846 TI - [Pre-operative chemotherapy in non-small cell lung cancer]. AB - Pre-operative chemotherapy for non-small cell bronchial carcinoma (NSCLC) has the twin objectives of destruction of micrometastases and increased resectabilty of the primary tumour. The trials of Rosell and Roth showed weak benefits in favour of the combination. In the French trial overall survival was no different for the whole group but was improved in early stage disease. Preliminary results of the SWOG trial show a non-significant difference in 2 year survival of 6%. The EORTC 08941 trial showed no difference between surgery and radiotherapy following induction chemotherapy in non-resectable stage IIIAN2 NSCLC. The INT-0139 trial compared surgery following induction chemo-radiotherapy with chemo-radiotherapy alone. There was no difference between the two strategies but analysis of sub groups suggested that some groups might benefit from the triple combination. Two further trials await publication. The small number of patients in each trial suggests that a meta-analysis will be necessary to reach a definite conclusion. The combination of surgery and chemotherapy is becoming standard in stage II disease. Only the timing, pre- or post-operative, remains controversial. At present, of the original objectives, only the destruction of micro-metastases has been confirmed. PMID- 16340847 TI - [Adjuvant chemotherapy for non-small cell lung cancer--which agents for which patients?]. AB - A meta-analysis ten years ago showed a modest non-statistically significant benefit for cisplatin-based chemotherapy in the adjuvant setting following resection of non-small cell lung cancer. Since then, several trials have evaluated this strategy. The IALT study measured the impact of cisplatin-based therapy, in association with a range of other agents versus observation and concluded positively, showing a modest but statistically significant improvement in survival. By contrast, the Italian ALPI study using MVP was negative. In the past two years, three major studies have been presented with clear positive results. Two of them, based on a Navelbine-cisplatin combination demonstrated very positive results in stages II and IIIA. A US study was positive in stage IB using Taxol and Carboplatin, in contrast to the Navelbine-cisplatin trials. Finally, the use of UFT, an oral fluoropyrimidine in Japan has also shown benefit in several trials confirmed by a meta-analysis. Based on the present recent trials, Navelbine-cisplatin should be considered after resection of stages II and IIIA NSCLC to improve the prognosis of those patients. PMID- 16340848 TI - [Non-small cell bronchial carcinoma: optimisation of radiotherapy]. AB - INTRODUCTION: Radiotherapy is involved in several ways in the treatment of non small cell bronchial carcinoma (NSCBC). BACKGROUND: Three dimensional radiotherapy, with or without chemotherapy, is current practice, radiotherapy with respiratory gating and with intensity modulation are being evaluated Developments in imaging allow more precise definition of target volume, and stereotactic irradiation and proton therapy are little used at present but very promising. VIEWPOINT: The level of local control of NSCBC remains unsatisfactory. The results will certainly improve in association with advances in three areas: chemotherapy, better dose escalation and more precise targeting of tumour volume. CONCLUSION: Improvement in the management of NSCBC depends on these new treatment modalities that are still being evaluated and improved, with the aim of obtaining a longer survival and a better quality of life with reduced toxicity and side effects. PMID- 16340849 TI - [Chemotherapy for metastatic non-small cell lung cancer]. AB - Treatment of stage IV non-small cell lung cancer (NSCLC) is based upon chemotherapy, with several meta-analyses showing an improvement of survival compared to supportive care alone. For first-line chemotherapy, the development of new third generation drugs (vinorelbine, gemcitabine, paclitaxel, docetaxel) with a better therapeutic index has led to the definition of 4 to 6 cycles of a platinum-based dual agent regime as standard therapy for performance stage (PS) 0 1 patients, producing a 1-year survival in the region of 35%. The choice of agents is based upon the differences in the toxicity profile of platinum-based combinations and on patient co-morbidities. For frail patients (PS>2 and elderly patients), single-agent chemotherapy seems more appropriate than cisplatin-based combinations; carboplatin or non-platinum combinations might be more effective. Third generation cytotoxic agents have modestly improved survival for patients with stage IV NSCLC but should probably provide an increased number of long-term survivors. PMID- 16340850 TI - [Improved prognosis in non small cell lung cancer--myth or reality?]. AB - Some data suggest that survival in non small cell lung cancer (NSCLC) has improved in the last ten years. A comparison between large prospective therapeutic trials from 1979 to 2002 does show some progress but there are many biases, especially the Will Rogers effect, where stage migration due to new methods of diagnostic imaging and invasive staging procedures could improve actuarial survival in each stage. Prospective randomised trials comparing older and more recent treatments demonstrate some improvements, specifically perioperative chemotherapy in stages I-IIIA, chemo-radiation in unresectable stage III disease and new drugs in stage IV. However retrospective whole population studies fail to show a significant improvement in survival. These discrepancies could be explained by differences between whole populations and those selected for clinical trials. Moreover, the results of the therapeutic trials are not always applied in all unselected patients. In conclusion, NSCLC survival can be improved in a selected population. To reproduce this in the whole population all patients should be managed in accordance with the results of recent trials. PMID- 16340851 TI - [Reports from the annual meeting of the American Thoracic Society, San Diego, California, USA, 20-25 May 2005]. PMID- 16340895 TI - [Suspension of ART coverage: results of a survey in France]. AB - The objective of the study was to investigate the criteria of patient selection and ART cycle cancellation in public and private fertility centers in France. A specific questionnaire was sent to 105 fertility centers. 81 centers have answered. Age was used as the main selection criteria. Others factors to consider are the number of previous IVF attempts, ovarian response and FSH levels. Less than 10 percent of centers investigate antral follicle count. PMID- 16340896 TI - [Overview of cancellations between November 2004 and March 2005 at the Antoine Beclere hospital]. AB - We analyzed 229 cycles in our IVF program between November 2004 and April 2005. The cancellation rate was 25%, but ranged from 20% in women below 37 years, to 50% in women aged over 41. Between 38 and 40 years old, it was 45%. FSH levels are significantly raised in women cancelled. Most of cancellations occurred at day 8 (46%) or at day 10 (40%), mostly because of low number of growing follicles. The risk of cancellation was increased when women had several pejorative predictive factors. The predictive value of the whole follicular cohort homogeneity is now being studied. It should be an important predictive factor of ovarian response to next stimulation cycle. PMID- 16340897 TI - [Analysis of the cancellations and temporary suspensions: the FIVNAT data]. AB - This article reports the results of the FIVNAT database concerning the cancellation of cycles in IVF programs. 8,980 cycles have been cancelled (11,5%), mostly because of a poor ovarian response to stimulation (66%). The chances of success of a second cycle of IVF following a cycle cancelled for poor ovarian response halved by 2 (compared with patients cancelled for HSO or for personal reasons). PMID- 16340898 TI - [What is the validity limit of patient reimbursement? Is age a criterion for refusal? Yes....]. AB - Selection is probably necessary for women undergoing ART (Assisted Reproductive Therapy), since cost of technique is very important. Furthermore, the right to try to achieve a pregnancy is a debatable issue, since the delivery rates are very low for ageing women. Success rates decrease strongly for women over 40, and women must be informed of it. Women 40 years or older are significantly less likely to achieve pregnancy with IUI as compared to women under age 35. The older women are also significantly less likely to achieve pregnancy with in vitro fertilization (IVF and IVF-ICSI). Costs per cycle were similar, however, for both groups. Moreover, older women utilizing assisted reproductive techniques are four to five times less likely to achieve normal delivery than the younger group, since pregnancy loss rate is greater. This poor prognosis for success in older women adds significantly to the mean cost per pregnancy as compared to younger women. Now, public institutions advocate and impose an age limit upon women who which to undergo ART. Ovulation induction should not be administrated for women over 45 years old (AFSSAPS, 2003). One should recommend not to propose to women over 42 years old and to men over 59 years old (Guidelines, 2004). Moreover, ART techniques cannot be reimbursed for women over 43 years old (CCAM, 2005). PMID- 16340899 TI - [What is the validity limit of patient reimbursement? Is age a criterion for refusal? No...]. AB - The late 20th century trend to delay birth of the first child until the age at which female fecundity or reproductive capacity is lower has increased the incidence of age-related infertility. In the female, the number of oocytes decreases with age until the menopause. Oocyte quality also diminishes, due in part to increased aneuploidy. A significantly lower pregnancy rate per stimulation and delivery rate per retrieval is found in women aged 40 to 45 years when compared with women aged 25 to 39 years. However, acceptable pregnancy can be achieved in woman aged 40 to 43 years (4 to 8%). Furthermore, very few women aged over 40 ask for IVF procedure. PMID- 16340900 TI - [FSH is a limiting factor]. AB - FSH concentrations obtained early in the follicular phase is a good predictive factor of ovarian response for assisted reproductive treatment. Nevertheless, FSH levels should be correlated with estradiol levels, antral follicle count and patient age. A history of elevated basal FSH levels in patients under the age of 40 years predicts a lower oocyte yield in IVF cycles but does not translate to either lower pregnancy or implantation rates. In women with uncertain ovarian function, antral follicle count and serum antimullerian hormone/mullerian inhibiting substance appear to be more discriminatory markers of assisted reproductive technology outcome. PMID- 16340901 TI - [Is the concentration of plasma FSH a criterion for refusal?]. AB - FSH level is correlated with ovarian reserve. Oocyte quality is correlated with women age. Pregnancy rate decreases and miscarriage rate increases with woman age. In young women, high FSH level are not associated with a lower pregnancy rates. Thus, in young women, high basal FSH is not a contraindication to IVF treatment. Miscarriage rate is not correlated with FSH level. The outcome of IVF is higher in young patients with elevated FSH levels, than in patients older than 40 years of age with normal FSH levels. PMID- 16340902 TI - [Uterine factors limiting ART coverage]. AB - Transvaginal sonographic evaluation of endometrial texture and thickness may be an indicator of the likelihood of achieving pregnancy. Very few pregnancies occur when the endometrial thickness was less than 6 mm. When type C endometrium (multilayered endometrium consisting of prominent outer and midline hyperechogenic lines and inner hypoechogenic regions) greater than or equal to 6 mm thick was seen, the pregnancy rate per embryo transfer is goods. MR imaging is highly accurate in helping to diagnose adenomyosis which is a cause of repeated implantation failures in women undergoing IVF. Furthermore the MRI evaluation of junctional zone thickness (AJZ>7 mm and MJZ>10 mm) is the best negative predictive factor of implantation failure (97%). Thus, we advocate the measurement of junctional zone thickness with MRI in women with 2 consecutive implantation failure following IVF. PMID- 16340903 TI - [The spermiological limits to ART]. AB - Spermatic limitations impairing IVF results exist. Globozoospermia is a severe form of teratozoospermia characterized by round-headed sperm with an absence of acrosome. This morphologic abnormality should decrease fertilization failure. Sperm DNA fragmentation is known to compromise male fertility. Furthermore previous findings have suggested the implication of oxidative stress in the aetiology of this pathological condition. Other studies have indicated that abnormally low parameters of sperm DNA integrity and sperm membrane integrity correlate to reduced fertility due in part to implantation disorders. Sperm DNA damage should be efficiently treated with oral antioxidants. Moreover, testicular sperm for use in intracytoplasmic sperm injection (ICSI) should be injected without delay. PMID- 16340904 TI - [Oocyte and embryo quality: is their morphology a good criterion?]. AB - This article evaluate the predictive value for implantation and pregnancy rates in IVF of the morphology of oocyte, zygote and embryo. Oocyte morphology does not affect fertilization rate, embryo quality and implantation rate after IVF/ICSI. Multinucleation in normally fertilized embryos is associated with lower implantation and pregnancy rates in in vitro fertilization-embryo transfer cycles. Severe blastomere fragmentation (>25%) is associated with lower delivery rates (1%). Secretion of human leukocyte antigen-G (sHLA-G) by human embryos is associated with a higher in vitro fertilization pregnancy rate. PMID- 16340905 TI - [Your ART ticket is no longer valid after how many attempts?]. AB - The economic implications of the use of assisted reproductive techniques require consideration. Since March 2005, in France, only 6 intrauterine inseminations and 4 IVF attempts are free of charge for the patient. However, definition of "acceptable" pregnancy rate for a new attempt remains discussed (10%, 15%, 20%?). Furthermore, several criteria should be considered when discussing a new IVF attempt, including woman age, FSH levels, antral follicle count and infertility aetiology. For example, in patients with secondary infertility, reasonable pregnancy rates (15-20%) can be archived even after 6 or 7 failures. PMID- 16340906 TI - [Indications and results of oocyte donation in Spain]. AB - In Spain, oocyte donors are selected from anonymous, well-informed university students over 18 years of age, who give their informed consent in writing. Before being accepted as donors, the candidates' personal and family medical histories were taken and they were given a gynaecological examination, genital ultrasonography, and analysed for viral infections and caryotype. The donors received economic compensation of about 900 euros. Among candidates for oocyte donation, 75% abandoned or were rejected following medical examination. The pregnancy rate was higher after transfer of fresh embryos (synchronous donor) (56%) as compared with frozen-tawned embryo transfer (46%). Among Turner's syndrome patients, cardiovascular complications are potentially the most severe during pregnancy, such as the exacerbation of a pre-existing hypertension and the dissection of aortic aneurysms. Logistic factors such as the travel and time commitment involved were major reasons for non-satisfaction of donors. PMID- 16340907 TI - [Economic aspects of the selection of patients and temporary suspensions]. AB - Economic implications of the use of ART (assisted reproductive techniques) require consideration. Initial patient selection is performed in few infertility centers, according to several criteria: age, FSH levels, follicle antral count, previous IVF attempt. Once should recognize that patient selection is efficient in reducing cost of IVF. Nevertheless, drop-out occurring in the middle of a IVF cycle increase Baby cost. PMID- 16340908 TI - [What are the health scheme's responses to "out of the ordinary" requests in ART?]. AB - This communication reviews some ethical considerations concerning debated indications for ART (Assisted Reproductive Therapy). Several case reports are discussed concerning the risks of IVF for women presenting severe pathology, the ethical responsibilities of investigators, social control of IVF. PMID- 16340909 TI - [Yes and no: ethical principles of assisted reproduction]. AB - Functions, main features, and operating principles of Clinical Ethical Committee for Assisted Reproductive Therapy in Cochin Hospital are presented. The communication explains this ethical methodology. PMID- 16340910 TI - [When and why to say "no" to patients desiring ART? The "no" that brings relief]. PMID- 16340911 TI - [Physiological and pathological autoimmunity in neurological disease]. AB - INTRODUCTION: Over the last years, the functions of the immune system have radically been revised. It has been illustrated how the different responses of the innate defense and the adaptative system were intimately intricated. STATE OF KNOWLEDGE: Innate immunity is an elaborate strategy, switched on by an invariant receptor-based response that is able to develop a specific defense against some pathogens. Moreover, this innate immunity governs T- and B-cell-dependent adaptive immune response, mediated via dendritic cells whose maturation is controlled by immune specificity. The concepts of autoreactivity have also strongly progressed and the functions of the physiological autoimmune response have been highlighted. The notion of T- and B-cell self-antigens not only shapes the immune repertoire, but also the self-recognition process which is a tool for the control of the immune response itself. PERSPECTIVE: New concepts of the pathophysiology of autoimmune diseases have emerged from a better understanding of the immune response, balancing between an intrinsic deregulation of the immune system and system deficiency to mount an effective response against an initial injury. CONCLUSION: Of course, therapeutic strategies are challenged by these data. We should expect that control of several autoimmune processes will be achieved in a few years, e.g. in rheumatoid arthritis or Crohn's disease controlled with biotherapies. PMID- 16340912 TI - [Inflammation and ischaemic stroke: current status and future perspectives]. AB - INTRODUCTION: Inflammation may play an important role in acute ischemic stroke. Experimental and clinical data suggest that post-stroke inflammatory responses are complex cascade phenomena, which may have detrimental or beneficial effects on outcome. This review focuses on the current understanding of inflammation effector pathways in focal cerebral ischemia and on therapeutic perspectives. STATE OF ARTS/PERSPECTIVES: Somatic markers, such as admission body temperature or C-reactive protein, seem to be correlated with outcome, but data are heterogeous and contradictory, perhaps because their kinetics are complex during the first hours of a stroke. The most studied effectors of inflammation are cellular adhesion molecules--CAM (including integrins, selectins and members of the immunoglobulin superfamily), leucocytes, microglia, and blood brain barrier permeability. Evidence from animal data has lead to some clinical trials investigating anti adhesion molecules and hypothermia. However, current results remain disappointing. Recent experimental data also suggest a possible beneficial role of insulin and statins, which may be mediated by their effects on the inflammatory response. CONCLUSION: Inflammation is an important avenue of therapeutic research in acute stroke. A better understanding of the inflammatory pathophysiology may help to a better design of clinical trials. PMID- 16340913 TI - [Immunofixation compared to isoelectric focusing in the detection of oligoclonal bands in cerebrospinal fluid of multiple sclerosis patients]. AB - INTRODUCTION: Intrathecal immunoglobulins (Ig) synthesis, reflected by oligoclonal bands (OCBs) in cerebrospinal fluid (CSF) is observed in up to 90 percent of patients with clinically definite Multiple Sclerosis (MS). The gold standard laboratory test to establish the presence of OCBs in CSF of MS patients is isoelectric focusing (IEF). However, a quicker and less expensive method has been developed: immunofixation (IF). METHODS: The aim of this study was to compare these two methods carried out 74 CSF/serum pairs of MS, 103 CSF/serum pairs of subject controls and to determine their sensitivity and specificity. RESULTS: The agreement between results from IEF and IF was excellent (Kappa = 0.84). IEF sensitivity (78 percent) was not significantly different from that of IF (74 percent) (p = 0.3). Similarly, the specificity of IEF (93 percent) was not significantly different from that of IF (95 percent) (p = 0.2). CONCLUSION: IF is a semi automated method which is easier to perform than IEF and which appears to be as efficient as IEF. PMID- 16340914 TI - [Devic's neuromyelitis optica in Morocco. A study of 9-cases]. AB - INTRODUCTION: Optic neuromyelitis or Devic's syndrome associates optic neuritis and myelitis. It can have a monophasic or relapsing course. The limits of this entity remain controversial. METHODS: We analyzed the results of a retrospective series of Moroccan patients with Devic's syndrome defined on the basis of Wingerchuk's criteria. RESULTS: Six women and three men were retained for study; average age was 39 years. Three patients presented with a monophasic form and six with relapsing disease. CSF analysis was abnormal in five. Brain MRI was normal in eight. Signal abnormalities extended over more than three vertebrae in eight patients. Infections were associated with neurological features in half of the patients. DISCUSSION: Our results are in agreement with those of large series in the literature regarding the clinical, imaging, and laboratory findings and disease course. We also noted that infections were more frequently associated with Devic's syndrome in our patients. CONCLUSION: Our study reports the results of the first North African series of Devic's syndrome patients. We suggest that this syndrome is particular in our context due to the high frequency and diversity of associated infections. PMID- 16340915 TI - [Interferon beta1-a IM once weekly does not modify general psychological function in the first months after therapy in patients with relapsing-remitting multiple sclerosis]. AB - INTRODUCTION: Psychological troubles are common in multiple sclerosis but their underlying etiology is still controversial. METHODS: The objective of this open, non comparative, multicenter study was to assess changes in global psychological functioning in new multiple sclerosis patients during the first 3 months of treatment with intramuscular interferon beta-1a once weekly (Avonex). This functioning was rated every 4 weeks with the GAF (Global Assessment Functioning) scale. Depression measured on MADRS (Montgomery & Asberg Depression Rating Scale), clinical global impression (CGI) on patients'psychological status and clinical as well as biological tolerance were also assessed every 4 weeks. RESULTS: Five hundred and ninety-nine patients (71.4 percent women), aged 39.4 years were included. No clinically significant difference in mean GAF score between baseline and the end of the first 3 months of interferon beta-1a IM treatment (main evaluation outcome) was found. Similar results were obtained on MADRS scale. CONCLUSION: No clinically significant alteration of global psychological functioning, including symptoms of depression, was observed during the first 3 months of treatment with interferon beta-1a IM. PMID- 16340916 TI - [Validity of the five-word screening test for Alzheimer's disease in a population based study]. AB - INTRODUCTION: In general medicine lack of time impairs screening for Alzheimer's disease (AD). The five word test (FWT) enables rapid assessment of verbal episodic memory in accordance with Grober and Buschke neuropsychological concept. The main steps of the FWT are: induce specific semantic processing, control of encoding to avoid attention deficits, free and cued recall. Cued recall helps to distinguish a recall impairment from storage impairment which is evocative of AD. OBJECTIVE: Evaluate FWT total score (sum of free and cued recalls), FWT total weighed score which give a higher coefficient for free recalls than cued recalls and present the ability of these two scores for AD screening. METHOD: Evaluation performed with 4116 subjects (of whom 73 MA) aged from 65 years and more, randomly selected in two French towns for the "Three Cities" Study, a population based cohort. RESULTS: The total score was more specific than sensitive with a maximal sensitivity (Se) at 63 percent with specificity (Sp) at 91.1 percent. The total weighed score significantly increased Se (83.6 percent) with control of specificity (84.9 percent) and positive predictive value (9.1 percent). CONCLUSION: The FWT allows quick screening of patients for whom further neuropsychological evaluation is needed to diagnose AD. The ability of is simple test to screen for AD is improved by a simple weighting procedure: the total weighted score. PMID- 16340917 TI - [Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) in patients with solid organ transplantation: a clinical, neurophysiological and neuropathological study of 4 cases]. AB - INTRODUCTION: Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) rarely develops in patients with solid organ transplantation. PATIENTS AND METHOD: We describe the clinical, biological, electrophysiological and neuropathological features of 4 patients with solid organ transplantation who developed CIDP. Two patients had liver transplantation, one had kidney transplantation and one had lung transplantation. RESULTS: All 4 patients developed in the months following transplantation a syndrome that fulfilled criteria for definite CIDP. All patients had immunosuppressive therapy, with ciclosporin + prednisolone in 2 cases, tacrolimus in 1 case and azathioprine + prednisolone + ciclosporin in one case. One patient had chronic HCV and HBV infection. Treatment with intravenous immune globulin (IVIG) and/or a change in immunosuppressive therapy improved the neuropathy in all cases. CONCLUSION: CIDP is a rare and potentially treatable condition that should be considered in all patients with solid organ transplantation who develop a rapidly disabling sensorimotor polyneuropathy. PMID- 16340918 TI - [Severe visual loss due to retinal artery occlusion associated with Susac's syndrome: a case report]. AB - INTRODUCTION: Susac's syndrome consists of the clinical triad of cerebral microangiopathy, branch retinal artery occlusions, and hearing loss. The pathogenesis of the disease remains unknown. The severity of retinal vasculopathy remains variable. CASE REPORT: A 57-year-old Caucasian woman presented in 1999 for the diagnostic and therapeutic management of a severe and complete form of the disease. Fluorescein angiography and cerebral MRI were contributive in this case and confirmed the diagnosis. A severe unilateral visual loss occurred one month after the initiation of systemic corticosteroids. Funduscopy disclosed a temporal retinal artery occlusion with major ischemia. Therefore, cyclophosphamide and anticoagulants were added to corticosteroids and total visual recovery was achieved within 2 weeks without any relapse after a follow-up period of 54 months. CONCLUSION: Therapeutic management of Susac's syndrome is still controversial. Major immunosuppressive regimens are mandatory in the face of severe visual loss associated with central or branch retinal artery occlusions. The final prognosis of the disease seems favorable in the absence of prolonged and symptomatic retinal ischemia. PMID- 16340919 TI - [Aseptic meningoencephalitis in primary Gougerot-Sjogren's syndrome]. AB - INTRODUCTION: Central nervous system involvement is rarely observed in primary Sjogren's syndrome (PSS) and aseptic meningoencephalitis occurs exceptionally. OBSERVATION: We report the case of a 25-year-old woman who presented with weakness of the four limbs, vision loss and concentration deficit. Neurological examination revealed a mental confusion, cerebellar involvement and spastic tetra paresis. Cerebrospinal fluid analysis revealed elevation of cell counts (162/mm3) and protein content (3 g/l). MRI showed increased T2 signals in the white matter. No evidence for an infectious etiology, especially tuberculosis, was obtained in this patient. PSS was diagnosed on the basis of ocular dryness, lachrymal hyposecretion, aspects of labial gland biopsy witch revealed destruction of the ducts with periductal lymphocytic infiltration and positive anti-SSB antibodies. Oral corticosteroids (1 mg/kg/day) allowed an improvement of the neurological manifestations. CONCLUSION: Aseptic meningoencephalitis can be the first manifestation of primary Sjogren's syndrome. Infectious causes, particularly tuberculosis which is frequent in our context must be ruled out. PMID- 16340920 TI - [Autonomic nervous system involvement in chronic inflammatory demyelinating polyneuropathy]. AB - INTRODUCTION: Involvement of the autonomic nervous system during Chronic Inflammatory Demyelinating Polyneuropathy (CIDP) is rare. Localised hyperhydrosis and Claude Bernard Horner (CBH) syndrome have never been reported in patients with CIDP. CASE REPORT: We report the case of a 65-year-old right handed man who presented with hyperhydrosis localised to the right hemithorax and hemiface and a left CBH syndrome. After an extensive workup, the patient was found to have CIDP as the only cause of autonomic nervous system involvement. The symptoms resolved slowly after three courses of intravenous immunoglobulins. CONCLUSION: Signs of autonomic nervous system involvement can be observed in CIDP as in Guillain-Barre syndrome. This case report shows that immunomodulatory treatment can be effective against dysautonomia in CIDP. PMID- 16340923 TI - [Electrophysiological signs of muscle channelopathies by causal mutation]. AB - Nondystrophic myotonias and familial periodic paralyses are disorders of muscle excitability caused by mutations in genes coding for voltage-gated ion channels. Electromyographic (EMG) diagnosis of these diseases is based on needle searching of myotonic discharges and on several exercise tests, performed by surface recording of compound muscle action potential (CMAP) before and after voluntary contractions of various durations. After a short exercise test (10 seconds contraction), three abnormal changes of CMAP have been defined in muscle channelopathies: 1) appearance of repetitive discharges (post-exercise myotonic potentials, PEMP), 2) transient or long-lasting amplitude reduction, 3) long lasting amplitude increase. These abnormalities become more marked or decline with exercise repetition (repeated brief exercise test), depending on the causal mutation. After long exercise test (5 minutes contraction), three other abnormalities of CMAP amplitude have been described: 1) immediate increase, 2) immediate decrease, 3) progressive or delayed decrease. Combining the responses to the different tests has disclosed five main EMG types of muscle channelopathies (I to V), that correlated with different pathophysiological mechanisms and with subgroups of mutations. This classification may guide molecular diagnosis in clinical practice. PMID- 16340924 TI - [Cramp-fasciculation syndrome]. AB - The cramp-fasciculation syndrome is a rare clinical entity in comparison with the frequency of cramps and isolated fasciculations in the general population. It is recognized as a benign syndrome without weakness and atrophy, however a few reports suggest that it may precede the occurrence of a motor neuron disease. Most often, the cramp-fasciculation syndrome is idiopathic and may be a component of a hyperexcitable peripheral nerve syndrome including other activities such as myokymia and neuromyotonia where antibodies to voltage-gated potassium channels (VGKCs) appear to be one of the effector mechanisms. The most complete form of this hyperexcitable peripheral nerve syndrome is Isaacs' syndrome. The central nervous system is also concerned with anti-VGKC antibodies found in Morvan's disease and limbic encephalitis which is often a paraneoplastic condition. These findings extend the spectrum of the anti-VGKC syndrome that may be associated with other auto-immune diseases, chiefly myasthenia gravis with thymoma. Carbamazepine and phenytoin cause reduction of the clinical and electrophysiological signs of the nerve hyperexcitability, and plasmapheresis and (or) immunosuppressors are useful when an auto-immune origin is considered. PMID- 16340925 TI - [Neuromuscular abnormalities in critical illness]. AB - Critical illness neuromuscular abnormalities (CINMA) are found in 25 percent of ITU patients who recover consciousness and are characterized by a bilateral and symmetric weakness that involves the four limbs but spares the facial muscles. Electrophysiological testing shows an axonal sensory motor polyneuropathy and/or myopathy. The main risk factors of CINMA are prolonged durations of multiple organ failure and mechanical ventilation, use of corticosteroids and hyperglycaemia. CINMA contribute also to increase the duration of mechanical ventilation, this effect being mediated by diaphragm weakness. The median duration of limb weakness is 21 days, although it can exceed several months in some patients. Few preventive measures have been assessed. Whether the benefit of strict blood glucose control in ITU patients recovering from heart surgery on CINMA incidence can be extended to medical ICU patients needs to be determined. PMID- 16340926 TI - [News on atopic dermatitis: new nomenclature, genetics and natural evolution]. PMID- 16340927 TI - [The influence of Avene thermal spring water on intra-cellular keratinocyte mobilisation]. PMID- 16340928 TI - [The hydrotherapy centre in Avene-les-bains. A controlled study in atopic dermatitis]. PMID- 16340929 TI - [The efficacy of Avene thermal spring water in light to moderate atopic dermatitis]. PMID- 16340930 TI - [Atopic dermatitis: the impact of hydrotherapy on quality of life]. PMID- 16340932 TI - Anemia: a modifiable risk factor for heart disease. Introduction. PMID- 16340933 TI - Piecing together the evidence on anemia: the link between chronic kidney disease and cardiovascular disease. AB - Chronic kidney disease (CKD) is now accepted as an independent cardiovascular disease (CVD)-risk state, regardless of its underlying cause. Anemia is a common feature of CKD, particularly in end-stage renal disease. Anemia is also independently associated with poorer outcomes in a wide variety of CVD states, including congestive heart failure and coronary artery disease. Anemia appears to act as an independent mortality multiplier when hemoglobin levels drop below 12 g/dL. With the independent and profound contribution of both CKD and anemia to cardiovascular mortality and morbidity, understanding the pathophysiologic links among these disease states is important. In addition, it is hoped that treatments currently under active investigation and geared specifically to attenuate the cardiovascular risk associated with anemia and CKD, such as erythropoietin therapy, will improve outcomes. This article reviews the evidence for an association among CKD, anemia, and CVD. PMID- 16340934 TI - Pathogenesis of anemia in cardiorenal disease. AB - The link between chronic heart failure (CHF) and chronic kidney disease (CKD) is well known. Approximately 50% of patients with CHF have some renal dysfunction, and 25% of patients with CKD and serum creatinine levels ranging from 1.5 mg/dL to 6 mg/dL have CHF. The association of CHF with CKD is strong and may contribute to its long-term progression. Anemia is also common in patients with CKD and contributes to increased morbidity and mortality. More recently, anemia has been found to be frequently present in patients with CHF, and its presence is associated with worse long-term CHF outcomes. Thus, anemia, CHF, and CKD may be independently related to one another, and this relationship may have important implications for their management. PMID- 16340935 TI - Arterial homeostasis, inflammation, and erythropoietic growth factors. AB - A neurohumoral link between kidneys and the heart has been established, particularly in the context of hypertension and cardiomyopathy. Beyond this neuro endocrine pathway, another connecting system theoretically recruits growth factors that are selectively produced by the kidneys and have the ability to promote a distant reaction at the level of bone marrow. This reaction differentiates and circulates vascular progenitor cells capable of repairing the injured cardiovascular system. Reducing injuries (prevention) stabilizes disease processes by reducing tissue damage and destruction but the gradual degradation of the body's natural repair mechanisms eventually allows progressive reactivation of disease processes. In this light, a focus on tissue repair rather than injury prevention may hold the key to controlling chronic heart diseases. This article examines the medical therapies, including recombinant human erythropoietin, that have been shown to improve the function and survival of endothelial progenitor cells and promote the healing of damaged tissue. PMID- 16340936 TI - Myocardial disease, anemia, and erythrocyte-stimulating proteins in chronic kidney disease. AB - The combination of heart failure and chronic kidney disease (CKD) has received comparatively little attention in terms of clinical research versus investigations of each state individually. It has been known for over a decade that anemia, a cardinal feature of CKD, is associated with higher cardiovascular event rates in late-stage and end-stage renal disease. Although the biological mechanisms linking anemia, renal failure, and heart failure are incompletely understood, more prevalent anemia is consistent in patients with more severe heart failure and is associated with higher mortality rates. Impaired erythropoietin production and resistance to erythropoietin are major contributors to anemia in patients with heart failure. By targeting hemoglobin levels in anemic patients with CKD, through the use of recombinant erythropoietin (epoetin) therapy, it has been hoped that anemia, CKD, and heart failure outcomes can be improved. Darbepoetin alfa was engineered to contain more N-linked carbohydrate chains than erythropoietin, and has an approximately 3 times longer serum half life. Several clinical trials have addressed the hypothesis that darbepoetin alfa can effectively treat renal anemia at dose frequencies of once per week, or less often, with positive outcomes. PMID- 16340937 TI - Uncertainty in the treatment of anemia in chronic kidney disease. AB - The practice of medicine requires that the assimilation of today's best available data be directed toward individual patient-care decisions. In the absence of definitive data, surrogate measures are adopted to anticipate how therapeutic strategies would influence clinical outcomes and prognosis. Unfortunately, randomized controlled clinical trials (RCTs) have not always found these estimates of outcome to be reliable. Anemia is a clear marker of adverse prognosis, which can be modified by erythropoietic stimulating proteins (ESP). At present, there is sufficient uncertainty regarding the risks and benefits of ESP treatment in patients with chronic kidney disease and anemia to warrant major RCTs. This article reviews the rationale and design features for these trials. PMID- 16340938 TI - Measles--United States, 2004. AB - Measles is a highly infectious, acute viral illness that can cause severe pneumonia, diarrhea, encephalitis, and death. During 2004, a total of 37 cases (incidence: <1 case per million population) was reported to CDC by local and state health departments, the lowest number of measles cases ever reported in 1 year in the United States and a decrease of 16% from the previous low of 44 cases in 2002. This report describes the epidemiology of measles in the United States in 2004, documenting the absence of endemic measles and the continued risk for internationally imported measles cases that can result in indigenous transmission. PMID- 16340939 TI - Late relapse of Plasmodium ovale malaria--Philadelphia, Pennsylvania, November 2004. AB - Approximately 1,300 cases of malaria are reported each year in the United States; nearly all of these cases occur in travelers, many of whom fail to receive or adhere to prescribed chemoprophylaxis or do not follow recommendations for prevention of mosquito bites. Malaria can persist if not treated or if treated incorrectly (e.g., with an ineffective drug or an incorrect dosage of an effective drug). Early treatment is required to avoid severe illness or death. Although malaria typically becomes clinically apparent within 1 month of infection, cases can occur years after the last presumed exposure. In November 2004, CDC received a report of a late relapse of malaria in a Nigerian man aged 23 years in Philadelphia, Pennsylvania. His malaria was determined to have been caused by Plasmodium ovale, one of the four species of Plasmodium parasite that are transmitted by mosquitoes and cause malaria. The patient had been treated for malaria in Nigeria on multiple occasions, most recently 6 years before onset of his illness in the United States. This report describes the Philadelphia case, which underscores the importance of taking a detailed travel and immigration history when evaluating unexplained fever and considering malaria in the differential diagnosis. PMID- 16340940 TI - Outbreak of cutaneous Bacillus cereus infections among cadets in a university military program--Georgia, August 2004. AB - Although Bacillus cereus is known mainly as an agent of food poisoning, other infections caused by this organism have been documented in immunocompromised patients, including sepsis, meningitis, pneumonia, and wound infections. Certain populations are at increased risk for B. cereus infection, including cancer patients, neonates, intravenous drug users, and patients with a history of trauma, surgery, or catheterization. Primary cutaneous disease attributed to B. cereus in immunocompetent persons or in non-health-care settings rarely has been reported. This report is the first to document such an outbreak. On August 24, 2004, a local health department in Georgia received a call from a university health center describing 90 cadets with nonpruritic, impetigo-like lesions on their scalps; B. cereus was the common organism among the three patients whose lesions were cultured. The cases occurred during the freshman military orientation week that preceded the start of the fall term. The Georgia Division of Public Health (GDPH) conducted an investigation to determine the source of the infections, identify associated risk factors, and implement control measures. This report summarizes the results of the outbreak investigation, which identified receiving a short haircut at the start of orientation week, sharing sunscreen during the week, and membership in Company B as strongly associated with having scalp lesions. Recommendations to the university included changing the type of haircut required, increasing time allowed for showering, and issuing individual sunscreen. The results of this investigation underscore the need for military programs to incorporate good hygiene and infection-control measures into school orientation events. PMID- 16340941 TI - Recommended antimicrobial agents for the treatment and postexposure prophylaxis of pertussis: 2005 CDC Guidelines. AB - The recommendations in this report were developed to broaden the spectrum of antimicrobial agents that are available for treatment and postexposure prophylaxis of pertussis. They include updated information on macrolide agents other than erythromycin (azithromycin and clarithromycin) and their dosing schedule by age group. PMID- 16340942 TI - Dietary intake in relation to self-reported constipation among Japanese women aged 18-20 years. AB - OBJECTIVE: Information on modifiable lifestyle factors associated with constipation is limited, especially among non-Western populations. We examined associations between dietary intake and self-reported constipation in young Japanese women. DESIGN: Cross-sectional study. SUBJECTS: A total of 1,705 female Japanese dietetic students aged 18-20 years and free of current disease and current dietary counseling. METHODS: Dietary intake was estimated over a 1-month period with a validated, self-administered, diet history questionnaire, and lifestyle variables including self-reported constipation were assessed by a second questionnaire designed for this survey. RESULTS: A total of 436 women (26%) reported themselves to be 'constipated'. A multivariate odds ratio (OR) for women in the highest quartile of rice intake was 0.47 (95% confidence interval (CI): 0.33, 0.68) compared with the lowest. Additionally, women in the highest category of coffee intake had a multivariate OR of 0.67 (0.47, 0.94) compared with women in the lowest. Conversely, women in the highest quartile of confectionery intake had a multivariate OR of 1.54 (1.12, 2.13) compared with women in the lowest. Moreover, a multivariate OR for constipation for women in the highest quartile of Japanese and Chinese tea intake was 1.49 (1.09, 2.05) compared with women in the lowest. Neither total dietary fiber intake nor other lifestyle factors examined were associated with constipation. CONCLUSIONS: The consumption of rice and coffee was inversely associated with and that of confectioneries and Japanese and Chinese tea was positively associated with a prevalence of self-reported constipation. PMID- 16340943 TI - Newborn thyrotropin screening confirms iodine deficiency in Latvia. AB - OBJECTIVE AND SETTING: The national neonatal thyrotropin (TSH) screening data for 2000-2002 were analysed to assess the extent and severity of iodine deficiency in Latvia. RESULTS: Elevated newborn TSH (>5 mIU/l) decreased from 16.5 to 10.4% and 8.4% in 2000, 2001 and 2002, respectively. For each year and in all regions, the prevalence of elevated newborn TSH was above the cut-off level indicative for a public health problem. A positive correlation (r=0.82, P<0.05) existed between elevated newborn TSH and low urinary iodine excretion among a nationwide sample of school children in 2000. CONCLUSION: Latvia is iodine deficient, putting each new generation at risk of preventable brain damage. PMID- 16340944 TI - Effects of different dietary protein intakes on body composition and vascular reactivity. AB - OBJECTIVE: To assess the effects of a diet rich in protein of animal origin in comparison to one with a protein intake of about 15% of the total daily calories on body composition and arterial function. DESIGN: Randomized prospective study with parallel groups. Body weight (BW), blood pressure (BP), main parameters of carbohydrate and lipid metabolism, body mass composition by bioelectrical impedance analysis, forearm blood flow at rest and in the postischaemic phase by strain gauge plethysmography and flow-mediated dilation of the brachial artery by echography were measured at baseline and after 6 months of the dietary intervention. SUBJECTS: In total, 15 clinically healthy male volunteers, regularly performing a mixed training three times weekly for 90 min. INTERVENTION: The participants were randomly prescribed a diet with high (1.9 g/kg BW) or normal (1.3 g/kg BW) protein content. STATISTICAL ANALYSIS: Differences between means were evaluated by the t-tests for paired or unpaired data and by one way analysis of variance. The strength of correlation between variables was investigated by bivariate Pearson correlation. RESULTS: Serum cholesterol significantly decreased with both diets in comparison to baseline values, whereas BW was slightly but significantly reduced only by the high protein (HP) diet. No change was detected in BP and the other metabolic parameters. Body mass composition was not significantly modified by either diet. On the other hand, postischaemic flow-mediated dilation of the brachial artery was enhanced by the sole normal protein (NP) diet, whereas no change in the forearm blood flow, both at rest and in the postischaemic phase, was detected. CONCLUSIONS: These preliminary results indicate that HP diet was found to be not useful in increasing the muscle mass in comparison to a NP intake. In contrast to this, the latter diet seems to enhance the endothelial function of the arterial vessels with a more pronounced dilatation of the lumen in response to the increase in blood flow. PMID- 16340945 TI - Changes in lifestyle and total homocysteine in relation to MTHFR(C677T) genotype: the Inter99 study. AB - BACKGROUND: Reduction in total homocysteine (tHcy) may be clinically relevant in the prevention of cardiovascular disease (CVD) in the general population. OBJECTIVE: To examine the effects of changes in various lifestyle habits and lifestyle related biological CVD risk markers on changes in tHcy in relation to MTHFR(C677T) genotype. DESIGN: A 1 year follow-up study. SETTING: Copenhagen County, Denmark. SUBJECTS: Statistical analyses were based on a population-based sample of 915 men and women aged 30-60 years assessed to be at increased CVD risk at baseline and therefore offered lifestyle intervention and re-examination after one year. RESULTS: None of the studied lifestyle changes-- smoking, physical activity, dietary habits, and coffee, tea, and alcohol consumption-- was significantly associated with changes in tHcy, either overall, or in any of the MTHFR genotype subgroups. In addition, changes in tHcy did not differ between participants randomized to low- and high-intensity lifestyle intervention. However, the MTHFR TT genotype was associated with a significant decrease in tHcy compared with the CC/CT genotype in which an increase was observed. In addition, changes in tHcy were associated with changes in several of the biological CVD risk markers: weight, total cholesterol, HDL cholesterol, LDL cholesterol and systolic blood pressure. CONCLUSIONS: Our results indicate that tHcy may not be reduced by lifestyle changes; additionally, they suggest that tHcy may be related to biological CVD risk markers through a lifestyle independent pathway. PMID- 16340946 TI - Experience of 6 years with home enteral nutrition in an area of Spain. AB - OBJECTIVE: The wide spread use of long-term enteral nutrition and the substantive costs dictate a need to study outcome, clinical and epidemiological characteristics of these patients. The aim of our study was to analyze incidence and characteristics of a cohort of patients on HEN during 6 years, after our previous pilot study of 3 years. DESIGN: Prospective observational study. SETTING: Tertiary care. SUBJECTS: Between January 1999 and December 2004, all adult patients living in Valladolid West area who were discharged from the hospital on HEN were prospectively studied and followed up. INTERVENTIONS: Information for each patient was prospectively recorded by the dietitian of the team, and include age, sex, body mass index, tricipital skinfold, midarm circumference, underlying disease, exitus, dates of initiation and discontinuation of HEN, nutrient formula, mode of administration and complications of HEN. During HEN, physicians supervised the home patients and the patients themselves or their close relative, were asked to contact our nutrition team if any problem occurred. Finally the yearly incidence of HEN was calculated each year on the basis of the estimated population in our area of recruitment, assuming almost all HEN patients were reported. RESULTS: In our previous study, incidence of HEN in 1999 was patients 15 per 100,000 inhabitants, 21.3 in 2000 and decreased to 9.52 in 2001. In the new 3 years, the incidence remained in the mean levels of 1999 and 2000, the data were 17.1 per 100,000 inhabitants 26.5 in 2003 and 25.6 in 2004. The mean age of all patients was 56.4+/-17 years. The distribution of patients by diseases was; (43.8%) had a head and neck cancer, (26.8%) had human immunodeficiency virus infection, (9.6%) had a neurological disorders affecting swallowing (cerebrovacular accident and/or dementia), (4.7%) had diseases in digestive tract (fistulae, pancreatic disease, inflammatory bowel disease), (2.5%) had tumors in different locations with anorexia, (5%) had head trauma, and (7.6%) had one of several miscellaneous diseases inducing dysphagia or anorexia. HEN was administered via oral in 258 patients (70.6%) (group I), via a NGT in 95 patients (26.1%), a PEG in 9 patients (2.5%), and a jejunostomy in 3 patients (0.8%) (group II=107 patients). During the course of HEN, 12 patients had diarrhea (3.3%) and 8 (2.2%) constipation, and 4 vomiting (1.1%) that did not require cessation of HEN. No lung aspiration was detected. Hypernatremia (sodium >145 meq/l) appeared only in six cases (1.64%) and 12 cases of hyperglycaemia (3.28%). Ten patients (2.8%) reported a least one problem with the tube. The mean duration of HEN was 148.2+/-104 days. After the follow-up, 19 of the 365 patients (5.2%) had died, 346 (94.8%) were alive. In multivariant analysis, an independent factor associated with death was age (hazard ratio: 1.22; 95% CI: 1.06-1.39), adjusted by sex, route and diagnosis. CONCLUSIONS: HEN has a high incidence in our area and it is a valid and safe technique for nutrition support. PMID- 16340947 TI - Family-based behavioural treatment of obesity: acceptability and effectiveness in the UK. AB - OBJECTIVE: To assess the acceptability and impact of family-based behavioural treatment (FBBT) for childhood obesity in a clinical setting in the UK. DESIGN: Pre- and post-treatment assessment for four consecutive treatment groups. SETTING: Treatment groups took place at Great Ormond Street Hospital, London. PATIENTS: Participants were 33 families with obese (BMI >or=98th centile for age and sex) children aged 8-13 years. INTERVENTION: FBBT was delivered over 12 sessions. MAIN OUTCOME MEASURES: Overweight (percentage BMI), self-esteem, mood and eating attitudes were assessed before and after treatment; overweight was re assessed at 3-month follow-up for those who completed treatment. RESULTS: The FBBT programme materials translated easily to the UK setting and the programme was well-liked by participants. Twenty-seven out of 33 families (82%) completed the sessions. Children lost 8.4% BMI over the time of treatment, and this was maintained at 3-month follow-up. Self-esteem and depression improved significantly and there was no change in food pre-occupation, anorexia or bulimia on the ChEAT. CONCLUSIONS: These results establish that FBBT is feasible and acceptable in a clinical setting in Britain. They indicate that significant loss of overweight can be achieved using the programme without adverse psychological consequences. PMID- 16340948 TI - Alterations in serum levels of trace elements in tuberculosis and HIV infections. AB - OBJECTIVE: To evaluate serum concentrations of trace elements in tuberculosis (TB) patients with or with out human immunodeficiency virus (HIV) coinfection before and after anti-TB chemotherapy. SUBJECTS: A total of 155 TB patients, 74 of which were coinfected with HIV, and 31 healthy controls from Gondar, Ethiopia. METHODS: Serum levels of copper, zinc, selenium and iron were determined using an inductively coupled plasma mass spectrometer from all subjects at baseline and from 44 TB patients (22 with HIV coinfection) at the end of an intensive phase of anti-TB chemotherapy. RESULTS: Compared with the control group, the concentrations of iron, zinc and selenium were significantly lower (P<0.05) while that of copper and copper/zinc ratio was significantly higher (P<0.05) in the serum of TB patients. TB patients with HIV coinfection had significantly lower serum zinc and selenium concentrations and significantly higher copper/zinc ratio compared to that in TB patients without HIV coinfection (P<0.05). The serum concentration of zinc had significantly increased at the end of intensive phase of anti-TB chemotherapy in patients without HIV coinfection (P<0.05). An increase in serum selenium level was observed in TB patients with or without HIV coinfection after therapy. On the contrary, serum copper concentration and copper/zinc ratio declined significantly after anti-TB chemotherapy irrespective of HIV serostatus (P<0.05). CONCLUSIONS: The results indicate that TB patients have altered profile of trace elements in their sera. This warrants the need for further investigations so that strategies for trace elements supplementation can be planned in addition to their potential as diagnostic parameters in monitoring responses to anti-TB chemotherapy. PMID- 16340949 TI - Oligofructose promotes satiety in healthy human: a pilot study. AB - OBJECTIVE: The administration of a fermentable dietary fibre (oligofructose) in rats increases satietogenic gut peptides and lowered spontaneous energy intake. The aim of the study was to assess the relevance of those effects of oligofructose on satiety and energy intake in humans. DESIGN: Single-blinded, crossover, placebo-controlled design, pilot study. SUBJECTS: Volunteers included five men and five women aged 21-39 years, BMI ranging from 18.5 to 27.4 kg/m(2), were randomly assigned as described below. INTERVENTIONS: Subjects were included in two 2-week experimental phases during which they received either fibre (oligofructose (OFS)) or placebo (dextrine maltose (DM)); a 2-week washout period was included between crossover phases. In total, 8 g OFS or 8 g DM were ingested twice daily (16 g/day in total). Energy intake, hunger, satiety, fullness and prospective food consumption were assessed with analogue scales at the end of each experimental phase. RESULTS: During breakfast, OFS significantly increases the satiety (P=0.04) without any difference on other sensations as compared to DM treatment periods. After lunch, no significant differences are observed between treatment period. At dinner, OFS significantly increases satiety (P=0.04), reduces hunger (P=0.04) and prospective food consumption (P=0.05). The energy intake at breakfast and lunch are significantly lower (P=0.01, 0.03, respectively) after OFS treatment than after DM treatment. Total energy intake per day is 5% lower during OFS than in DM period. CONCLUSION: Oligofructose treatment increases satiety following breakfast and dinner, reduces hunger and prospective food consumption following dinner. This pilot study presents a rationale to propose oligofructose supplements in the management of food intake in overweight and obese patients. PMID- 16340950 TI - Effects of administration of iron, iodine and simultaneous iron-plus-iodine on the thyroid hormone profile in iron-deficient adolescent Iranian girls. AB - OBJECTIVE: To investigate whether iron supplementation can improve thyroid hormone function in iron-deficient adolescent girls. DESIGN: A double-blind randomized intervention study. SETTING: The study was performed from 2002 through 2003 in the Islamic Republic of Iran. SUBJECTS: 103 iron-deficient non-anaemic girls who fulfilled all inclusion criteria were included, and 94 subjects successfully completed the study. INTERVENTIONS: Patients were randomly assigned to one of four groups and treated with a single oral dose of 190 mg iodine plus 300 mg ferrous sulphate 5 times/week (n=24), 300 mg ferrous sulphate 5 times/week (n=23), a single oral dose of 190 mg iodine (n=25), or a placebo (n=22) for 12 weeks. RESULTS: All groups were comparable at baseline. After the intervention, there was a significant increase in ferritin and transferrin saturation in the iron+iodine group (17.6 vs 8.7 microg/dl, and 18.8 vs 7.2%, respectively, P<0.001 for both) and in the iron group (P<0.001 for both). Urinary iodine doubled in the iron+iodine group and in the iodine group (P<0.001 for both). Thyroid indices tT4, tT3 and T3RU increased and reverse RT3 decreased in the iron+iodine group (10 vs 8.9 microg/dl, P< 0.001; 143 vs 138 microg/dl, P<0.05; 32.3 vs 28.4%, P<0.001 and 24.8 vs 44.2 ng/dl, P<0.001, respectively) and in the iron group. These two groups did not differ for any of the four indices, but both differed significantly from the iodine and placebo groups. CONCLUSIONS: Our results indicate that improvement of iron status was accompanied by an improvement in some indices of thyroid hormones. SPONSORSHIP: This study was supported by the Dean of Research Affairs of the Tehran University of Medical Sciences. PMID- 16340951 TI - Within- and between-individual variability in estimated energy expenditure and habitual physical activity among young adults. AB - OBJECTIVE: To examine the within- and between-individual variability in the estimated total energy expenditure (TEE) and habitual physical activity (PA) in free-living young adults. We were specifically interested in the day-to-day variation. DESIGN: Cross-sectional. SETTING: University. SUBJECTS: A total of 277 (125 males, 152 females) participants aged 18-24 years. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Subjects recorded their daily PA with the Bouchard 3-day diary. Descriptive statistics for TEE, moderate-to-vigorous physical activity (MVPA), inactivity (IA), and activity energy expenditure (AEE) were calculated and presented as a frequency distribution. Within- and between-individual variability were tested by RMANOVA and also presented as the coefficient of variation (CV). RESULTS: Mean values for TEE, MVPA, AEE, and IA were comparable between males and females; however, considerable between-individual variability existed. Only MVPA showed a significant difference between sexes (P<0.05), due to slightly greater vigorous PA in males compared to females (8.0 and 6.8 kcal/kg/day, respectively; P=0.12). Results from the RMANOVA revealed no significant day-to-day variability in any TEE or PA variable and the CVs were not significantly different between males and females. TEE and IA displayed mean CVs of approximately 12% while mean CVs for AEE and MVPA approximated 32 and 84%, respectively. CONCLUSIONS: Results from this study suggest (1) a limited day-to-day variability in TEE and (2) a large degree of between-individual variation for TEE and PA exist. This study highlights the importance of understanding the variability associated with TEE, which is important for PA measurement and proper PA and nutritional recommendations. SPONSORSHIP: None. PMID- 16340953 TI - Effects of probiotics on the severity of experimental acute pancreatitis. AB - OBJECTIVE: This study was designed to evaluate the effects of probiotics on the severity of experimental acute pancreatitis. DESIGN: Experimental study. SETTING: Experiments were done in a laboratory at Haydarpasa Numune Teaching and Research Hospital. SUBJECTS: A total of 50 Wistar rats were randomly divided into five groups. INTERVENTIONS: Group 1 was control group. Group 2 received an intraperitoneal injection of a 20% solution in 0.15 mol/l NaCL. Group 3 was injected NaCL and fed with probiotics. Acute pancreatitis was induced in rats by intrperitoneal injection of L-Arginine in groups 4 and 5. The rats in group 5 were treated with probiotics. The pancreas was removed for histologic examination. Evaluation of the pathologic changes was done by a new combined histopathologic grading scale. RESULTS: The mean scores of fibrosis, acinar cell loss, oedema, parenchymal necrosis, mononuclear cells infiltration, polymorphonuclear leucocytes infiltration, ductal damage and atypical reactive regeneration in group 5 were significantly lower than group 4. CONCLUSIONS: We demonstrated that enteral feedings with added probiotics can reduce the severity of acute pancreatitis. SPONSORSHIP: None. PMID- 16340952 TI - Seasonal variation in food intake, physical activity, and body weight in a predominantly overweight population. AB - OBJECTIVE: To describe seasonal variation in food intake, physical activity, and body weight in a predominantly overweight population. DESIGN: A longitudinal observational study. SETTING: Most of the study participants were recruited from a health maintenance organization (HMO) in central Massachusetts, USA. Additional individuals of Hispanic descent were recruited from outside of the HMO population to increase the ethnic diversity of this sample. SUBJECTS: Data from 593 participants, aged 20-70, were used for this investigation. Each participant was followed quarterly (five sampling points: baseline and four consecutive quarters) for 1-year period. Body weight measurements and three 24-h dietary and physical activity recalls were obtained on randomly selected days (including 2 weekdays and 1 weekend day) per quarter. Sinusoidal regression models were used to estimate peak-to-trough amplitude and phase of the peaks. RESULTS: Daily caloric intake was higher by 86 kcal/day during the fall compared to the spring. Percentage of calories from carbohydrate, fat and saturated fat showed slight seasonal variation, with a peak in the spring for carbohydrate and in the fall for total fat and saturated fat intake. The lowest physical activity level was observed in the winter and the highest in the spring. Body weight varied by about 1/2 kg throughout the year, with a peak in the winter (P<0.001 winter versus summer). Greater seasonal variation was observed in subjects who were male, middle aged, nonwhite, and less educated. CONCLUSIONS: Although there is seasonal variation in diet, physical activity and body weight, the magnitude of the change is generally small in this population. SPONSORSHIP: US National Heart, Lung and Blood Institute. PMID- 16340954 TI - Dietary patterns and associated lifestyles in preconception, pregnancy and postpartum. AB - OBJECTIVE: To identify dietary patterns in women who are planning immediate pregnancy in preconception, weeks 6, 10, 26 and 38 of pregnancy, and 6 months postpartum, and to describe how particular lifestyles, the body mass index (BMI) and sociodemographic factors are associated to these patterns. DESIGN: Longitudinal study throughout the reproductive cycle of food consumption carried out in a Spanish Mediterranean city. SETTING: Faculty of Medicine and Health Sciences, Rovira i Virgili University. SUBJECTS: In total, 80 healthy female volunteers who were planning immediate pregnancy. INTERVENTIONS: A seven consecutive-day dietary record was used to evaluate the dietary intake. Exploratory factor analysis was used to identify the main dietary patterns in each of the periods. Fitted multiple linear regression models were used to study the associations between the lifestyle and sociodemographic variables, and each dietary pattern. RESULTS: The 'sweetened beverages and sugars' pattern was identified from preconception to 6 months postpartum and the 'vegetables and meat' pattern to the end of pregnancy. The 'sweetened beverages and sugars' pattern is positively associated with smoking and negatively associated with physical activity before conception and in the first trimester of pregnancy. The 'vegetables and meat' pattern is negatively associated with the BMI during the preconception period and positively associated with age in weeks 10 and 38 of pregnancy. It is shown that the patterns do not change significantly throughout the period studied. CONCLUSIONS: We have identified two stable dietary patterns from preconception to postpartum. The 'sweetened beverages and sugars' pattern is associated with habits of risk for the health of the pregnant woman and her offspring. SPONSORSHIP: 'Comision Interministerial de Ciencia y Tecnologia' (CICYT: ALI89-0388) and 'Instituto de Salud Carlos III', RCMN (C03/08), Madrid, Spain'. PMID- 16340955 TI - Bone morphogenetic protein signalling and vertebrate nervous system development. AB - Transforming growth factor-beta (TGFbeta) signalling, particularly signalling from the bone morphogenetic protein (BMP) members of this protein family, is crucial for the development of both the central and peripheral nervous systems in vertebrates. Experimental embryology and genetics performed in a range of organisms are providing insights into how BMPs establish the neural tissue and control the types and numbers of neurons formed. These studies also highlight the interactions between different developmental signals that are necessary to form a functional nervous system. The challenges ahead will be to uncover functions of TGFbeta signalling in later stages of CNS development, as well as to determine possible associations with neurological diseases. PMID- 16340956 TI - Self-motion-induced eye movements: effects on visual acuity and navigation. AB - Self-motion disturbs the stability of retinal images by inducing optic flow. Objects of interest need to be fixated or tracked, yet these eye movements can infringe on the experienced retinal flow that is important for visual navigation. Separating the components of optic flow caused by an eye movement from those due to self-motion, as well as using optic flow for visual navigation while simultaneously maintaining visual acuity on near targets, represent key challenges for the visual system. Here we summarize recent advances in our understanding of how the visuomotor and vestibulomotor systems function and interact, given the complex task of compensating for instabilities of retinal images, which typically vary as a function of retinal location and differ for each eye. PMID- 16340957 TI - International perspectives on engaging the public in neuroethics. AB - With an ever-increasing understanding of the brain mechanisms associated with core human attributes and values, there is an increasing public interest in the results of neuroscience research and the ways in which that new knowledge will be used. Here, we present perspectives on engaging the public on these issues on an international scale, the role of the media, and prospects for the new field of neuroethics as both a focus and a driver of these efforts. PMID- 16340958 TI - Vasoconstrictive neurovascular coupling during focal ischemic depolarizations. AB - Ischemic depolarizing events, such as repetitive spontaneous periinfarct spreading depolarizations (PIDs), expand the infarct size after experimental middle cerebral artery (MCA) occlusion. This worsening may result from increased metabolic demand, exacerbating the mismatch between cerebral blood flow (CBF) and metabolism. Here, we present data showing that anoxic depolarization (AD) and PIDs caused vasoconstriction and abruptly reduced CBF in the ischemic cortex in a distal MCA occlusion model in mice. This reduction in CBF during AD increased the area of cortex with 20% or less residual CBF by 140%. With each subsequent PID, this area expanded by an additional 19%. Drugs that are known to inhibit cortical spreading depression (CSD), such as N-methyl-D-aspartate receptor antagonists MK 801 and 7-chlorokynurenic acid, and sigma-1 receptor agonists dextromethorphan and carbetapentane, did not reduce the frequency of PIDs, but did diminish the severity of episodic hypoperfusions, and prevented the expansion of severely hypoperfused cortex, thus improving CBF during 90 mins of acute focal ischemia. In contrast, AMPA receptor antagonist NBQX, which does not inhibit CSD, did not impact the deterioration in CBF. When measured 24 h after distal MCA occlusion, infarct size was reduced by MK-801, but not by NBQX. Our results suggest that AD and PIDs expand the CBF deficit, and by so doing negatively impact lesion development in ischemic mouse brain. Mitigating the vasoconstrictive neurovascular coupling during intense ischemic depolarizations may provide a novel hemodynamic mechanism of neuroprotection by inhibitors of CSD. PMID- 16340959 TI - Evidence that stem cells reside in the adult Drosophila midgut epithelium. AB - Adult stem cells maintain organ systems throughout the course of life and facilitate repair after injury or disease. A fundamental property of stem and progenitor cell division is the capacity to retain a proliferative state or generate differentiated daughter cells; however, little is currently known about signals that regulate the balance between these processes. Here, we characterize a proliferating cellular compartment in the adult Drosophila midgut. Using genetic mosaic analysis we demonstrate that differentiated cells in the epithelium arise from a common lineage. Furthermore, we show that reduction of Notch signalling leads to an increase in the number of midgut progenitor cells, whereas activation of the Notch pathway leads to a decrease in proliferation. Thus, the midgut progenitor's default state is proliferation, which is inhibited through the Notch signalling pathway. The ability to identify, manipulate and genetically trace cell lineages in the midgut should lead to the discovery of additional genes that regulate stem and progenitor cell biology in the gastrointestinal tract. PMID- 16340960 TI - The adult Drosophila posterior midgut is maintained by pluripotent stem cells. AB - Vertebrate and invertebrate digestive systems show extensive similarities in their development, cellular makeup and genetic control. The Drosophila midgut is typical: enterocytes make up the majority of the intestinal epithelial monolayer, but are interspersed with hormone-producing enteroendocrine cells. Human (and mouse) intestinal cells are continuously replenished by stem cells, the misregulation of which may underlie some common digestive diseases and cancer. In contrast, stem cells have not been described in the intestines of flies, and Drosophila intestinal cells have been thought to be relatively stable. Here we use lineage labelling to show that adult Drosophila posterior midgut cells are continuously replenished by a distinctive population of intestinal stem cells (ISCs). As in vertebrates, ISCs are multipotent, and Notch signalling is required to produce an appropriate fraction of enteroendocrine cells. Notch is also required for the differentiation of ISC daughter cells, a role that has not been addressed in vertebrates. Unlike previously characterized stem cells, which reside in niches containing a specific partner stromal cell, ISCs adjoin only the basement membrane, differentiated enterocytes and their most recent daughters. The identification of Drosophila intestinal stem cells with striking similarities to their vertebrate counterparts will facilitate the genetic analysis of normal and abnormal intestinal function. PMID- 16340961 TI - Structural mechanism of plant aquaporin gating. AB - Plants counteract fluctuations in water supply by regulating all aquaporins in the cell plasma membrane. Channel closure results either from the dephosphorylation of two conserved serine residues under conditions of drought stress, or from the protonation of a conserved histidine residue following a drop in cytoplasmic pH due to anoxia during flooding. Here we report the X-ray structure of the spinach plasma membrane aquaporin SoPIP2;1 in its closed conformation at 2.1 A resolution and in its open conformation at 3.9 A resolution, and molecular dynamics simulations of the initial events governing gating. In the closed conformation loop D caps the channel from the cytoplasm and thereby occludes the pore. In the open conformation loop D is displaced up to 16 A and this movement opens a hydrophobic gate blocking the channel entrance from the cytoplasm. These results reveal a molecular gating mechanism which appears conserved throughout all plant plasma membrane aquaporins. PMID- 16340962 TI - Planetary science: are there active glaciers on Mars? AB - Head et al. interpret spectacular images from the Mars Express high-resolution stereo camera as evidence of geologically recent rock glaciers in Tharsis and of a piedmont ('hourglass') glacier at the base of a 3-km-high massif east of Hellas. They attribute growth of the low-latitude glaciers to snowfall during periods of increased spin-axis obliquity. The age of the hourglass glacier, considered to be inactive and slowly shrinking beneath a debris cover in the absence of modern snowfall, is estimated to be more than 40 Myr. Although we agree that the maximum glacier extent was climatically controlled, we find evidence in the images to support local augmentation of accumulation from snowfall through a mechanism that does not require climate change on Mars. PMID- 16340966 TI - Launching a business. PMID- 16340967 TI - Unapproved tests on a chip. PMID- 16340968 TI - Enough, already. PMID- 16340970 TI - UK animal labs still under siege. PMID- 16340971 TI - TV show gives research lobbyist a rat's-eye view of laboratory life. PMID- 16340972 TI - Animal-rights militancy exported to US and Europe. PMID- 16340973 TI - TV tests call into question cloner's stem-cell success. PMID- 16340975 TI - Avian flu centre put under threat of closure. PMID- 16340976 TI - Prospect of stricter timekeeping alarms US biologists. PMID- 16340977 TI - Climate talks edge towards twin-track future. PMID- 16340978 TI - Cowrie study strikes a blow for traditional taxonomy. PMID- 16340979 TI - German hostage was saving Iraq digs. PMID- 16340980 TI - Chairman explains Europe's research council. Interviewed by Alison Abbott. PMID- 16340983 TI - Neutron science: back on track? PMID- 16340982 TI - Nonsense mutations: running the red light. PMID- 16340984 TI - Fetal genetic testing: Screen test. PMID- 16340985 TI - Internet star shoots for a rocket revolution. PMID- 16340988 TI - Flu virus will not be sent in the regular US mail. PMID- 16340989 TI - Turkish science needs more than membership of the EU. PMID- 16340990 TI - Supplementary data need to be kept in public repositories. PMID- 16340995 TI - The message of the quantum. PMID- 16340996 TI - Genomics: the dog has its day. PMID- 16340997 TI - Water: ins and outs of ice nucleation. PMID- 16340999 TI - Quantum information: remember that photon. PMID- 16340998 TI - Cell biology: relays at the membrane. PMID- 16341000 TI - Cancer biology: emissaries set up new sites. PMID- 16341002 TI - Obituary: Alastair Cameron (1925-2005). PMID- 16341003 TI - Biomechanics: no force limit on greyhound sprint speed. AB - Maximum running speed is constrained by the speed at which the limbs can be swung forwards and backwards, and by the force they can withstand while in contact with the ground. Humans sprinting around banked bends change the duration of foot contact to spread the time over which the load is applied, thereby keeping the force on their legs constant. We show here that, on entering a tight bend, greyhounds do not change their foot-contact timings, and so have to withstand a 65% increase in limb forces. This supports the idea that greyhounds power locomotion by torque about the hips, so--just as in cycling humans--the muscles that provide the power are mechanically divorced from the structures that support weight. PMID- 16341005 TI - Astrophysics: is a doomsday catastrophe likely? AB - The risk of a doomsday scenario in which high-energy physics experiments trigger the destruction of the Earth has been estimated to be minuscule. But this may give a false sense of security: the fact that the Earth has survived for so long does not necessarily mean that such disasters are unlikely, because observers are, by definition, in places that have avoided destruction. Here we derive a new upper bound of one per billion years (99.9% confidence level) for the exogenous terminal-catastrophe rate that is free of such selection bias, using calculations based on the relatively late formation time of Earth. PMID- 16341008 TI - Measurement-induced entanglement for excitation stored in remote atomic ensembles. AB - A critical requirement for diverse applications in quantum information science is the capability to disseminate quantum resources over complex quantum networks. For example, the coherent distribution of entangled quantum states together with quantum memory (for storing the states) can enable scalable architectures for quantum computation, communication and metrology. Here we report observations of entanglement between two atomic ensembles located in distinct, spatially separated set-ups. Quantum interference in the detection of a photon emitted by one of the samples projects the otherwise independent ensembles into an entangled state with one joint excitation stored remotely in 10(5) atoms at each site. After a programmable delay, we confirm entanglement by mapping the state of the atoms to optical fields and measuring mutual coherences and photon statistics for these fields. We thereby determine a quantitative lower bound for the entanglement of the joint state of the ensembles. Our observations represent significant progress in the ability to distribute and store entangled quantum states. PMID- 16341007 TI - VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre metastatic niche. AB - The cellular and molecular mechanisms by which a tumour cell undergoes metastasis to a predetermined location are largely unknown. Here we demonstrate that bone marrow-derived haematopoietic progenitor cells that express vascular endothelial growth factor receptor 1 (VEGFR1; also known as Flt1) home to tumour-specific pre metastatic sites and form cellular clusters before the arrival of tumour cells. Preventing VEGFR1 function using antibodies or by the removal of VEGFR1(+) cells from the bone marrow of wild-type mice abrogates the formation of these pre metastatic clusters and prevents tumour metastasis, whereas reconstitution with selected Id3 (inhibitor of differentiation 3)-competent VEGFR1+ cells establishes cluster formation and tumour metastasis in Id3 knockout mice. We also show that VEGFR1+ cells express VLA-4 (also known as integrin alpha4beta1), and that tumour specific growth factors upregulate fibronectin--a VLA-4 ligand--in resident fibroblasts, providing a permissive niche for incoming tumour cells. Conditioned media obtained from distinct tumour types with unique patterns of metastatic spread redirected fibronectin expression and cluster formation, thereby transforming the metastatic profile. These findings demonstrate a requirement for VEGFR1+ haematopoietic progenitors in the regulation of metastasis, and suggest that expression patterns of fibronectin and VEGFR1+VLA-4+ clusters dictate organ specific tumour spread. PMID- 16341009 TI - Storage and retrieval of single photons transmitted between remote quantum memories. AB - An elementary quantum network operation involves storing a qubit state in an atomic quantum memory node, and then retrieving and transporting the information through a single photon excitation to a remote quantum memory node for further storage or analysis. Implementations of quantum network operations are thus conditioned on the ability to realize matter-to-light and/or light-to-matter quantum state mappings. Here we report the generation, transmission, storage and retrieval of single quanta using two remote atomic ensembles. A single photon is generated from a cold atomic ensemble at one site , and is directed to another site through 100 metres of optical fibre. The photon is then converted into a single collective atomic excitation using a dark-state polariton approach. After a programmable storage time, the atomic excitation is converted back into a single photon. This is demonstrated experimentally, for a storage time of 0.5 microseconds, by measurement of an anti-correlation parameter. Storage times exceeding ten microseconds are observed by intensity cross-correlation measurements. This storage period is two orders of magnitude longer than the time required to achieve conversion between photonic and atomic quanta. The controlled transfer of single quanta between remote quantum memories constitutes an important step towards distributed quantum networks. PMID- 16341010 TI - Electromagnetically induced transparency with tunable single-photon pulses. AB - Techniques to facilitate controlled interactions between single photons and atoms are now being actively explored. These techniques are important for the practical realization of quantum networks, in which multiple memory nodes that utilize atoms for generation, storage and processing of quantum states are connected by single-photon transmission in optical fibres. One promising avenue for the realization of quantum networks involves the manipulation of quantum pulses of light in optically dense atomic ensembles using electromagnetically induced transparency (EIT, refs 8, 9). EIT is a coherent control technique that is widely used for controlling the propagation of classical, multi-photon light pulses in applications such as efficient nonlinear optics. Here we demonstrate the use of EIT for the controllable generation, transmission and storage of single photons with tunable frequency, timing and bandwidth. We study the interaction of single photons produced in a 'source' ensemble of 87Rb atoms at room temperature with another 'target' ensemble. This allows us to simultaneously probe the spectral and quantum statistical properties of narrow-bandwidth single-photon pulses, revealing that their quantum nature is preserved under EIT propagation and storage. We measure the time delay associated with the reduced group velocity of the single-photon pulses and report observations of their storage and retrieval. PMID- 16341006 TI - Genome sequence, comparative analysis and haplotype structure of the domestic dog. AB - Here we report a high-quality draft genome sequence of the domestic dog (Canis familiaris), together with a dense map of single nucleotide polymorphisms (SNPs) across breeds. The dog is of particular interest because it provides important evolutionary information and because existing breeds show great phenotypic diversity for morphological, physiological and behavioural traits. We use sequence comparison with the primate and rodent lineages to shed light on the structure and evolution of genomes and genes. Notably, the majority of the most highly conserved non-coding sequences in mammalian genomes are clustered near a small subset of genes with important roles in development. Analysis of SNPs reveals long-range haplotypes across the entire dog genome, and defines the nature of genetic diversity within and across breeds. The current SNP map now makes it possible for genome-wide association studies to identify genes responsible for diseases and traits, with important consequences for human and companion animal health. PMID- 16341011 TI - A lithospheric instability origin for Columbia River flood basalts and Wallowa Mountains uplift in northeast Oregon. AB - Flood basalts appear to form during the initiation of hotspot magmatism. The Columbia River basalts (CRB) represent the largest volume of flood basalts associated with the Yellowstone hotspot, yet their source appears to be in the vicinity of the Wallowa Mountains, about 500 km north of the projected hotspot track. These mountains are composed of a large granitic pluton intruded into a region of oceanic lithosphere affinity. The elevation of the interface between Columbia River basalts and other geological formations indicates that mild pre eruptive subsidence took place in the Wallowa Mountains, followed by syn-eruptive uplift of several hundred metres and a long-term uplift of about 2 km. The mapped surface uplift mimics regional topography, with the Wallowa Mountains in the centre of a 'bull's eye' pattern of valleys and low-elevation mountains. Here we present the seismic velocity structure of the mantle underlying this region and erosion-corrected elevation maps of lava flows, and show that an area of reduced mantle melt content coincides with the 200-km-wide topographic uplift. We conclude that convective downwelling and detachment of a compositionally dense plutonic root can explain the timing and magnitude of Columbia River basalt magmatism, as well as the surface uplift and existence of the observed melt depleted mantle. PMID- 16341012 TI - Determinants of woody cover in African savannas. AB - Savannas are globally important ecosystems of great significance to human economies. In these biomes, which are characterized by the co-dominance of trees and grasses, woody cover is a chief determinant of ecosystem properties. The availability of resources (water, nutrients) and disturbance regimes (fire, herbivory) are thought to be important in regulating woody cover, but perceptions differ on which of these are the primary drivers of savanna structure. Here we show, using data from 854 sites across Africa, that maximum woody cover in savannas receiving a mean annual precipitation (MAP) of less than approximately 650 mm is constrained by, and increases linearly with, MAP. These arid and semi arid savannas may be considered 'stable' systems in which water constrains woody cover and permits grasses to coexist, while fire, herbivory and soil properties interact to reduce woody cover below the MAP-controlled upper bound. Above a MAP of approximately 650 mm, savannas are 'unstable' systems in which MAP is sufficient for woody canopy closure, and disturbances (fire, herbivory) are required for the coexistence of trees and grass. These results provide insights into the nature of African savannas and suggest that future changes in precipitation may considerably affect their distribution and dynamics. PMID- 16341013 TI - Morphine reward in dopamine-deficient mice. AB - Dopamine has been widely implicated as a mediator of many of the behavioural responses to drugs of abuse. To test the hypothesis that dopamine is an essential mediator of various opiate-induced responses, we administered morphine to mice unable to synthesize dopamine. We found that dopamine-deficient mice are unable to mount a normal locomotor response to morphine, but a small dopamine independent increase in locomotion remains. Dopamine-deficient mice have a rightward shift in the dose-response curve to morphine on the tail-flick test (a pain sensitivity assay), suggesting either a decreased sensitivity to the analgesic effects of morphine and/or basal hyperalgesia. In contrast, dopamine deficient mice display a robust conditioned place preference for morphine when given either caffeine or l-dihydroxyphenylalanine (a dopamine precursor that restores dopamine throughout the brain) during the testing phases. Together, these data demonstrate that dopamine is a crucial component of morphine-induced locomotion, dopamine may contribute to morphine analgesia, but that dopamine is not required for morphine-induced reward as measured by conditioned place preference. PMID- 16341014 TI - BMP inhibition-driven regulation of six-3 underlies induction of newt lens regeneration. AB - Lens regeneration in adult newts is a classic example of how cells can faithfully regenerate a complete organ through the process of transdifferentiation. After lens removal, the pigment epithelial cells of the dorsal, but not the ventral, iris dedifferentiate and then differentiate to form a new lens. Understanding how this process is regulated might provide clues about why lens regeneration does not occur in higher vertebrates. The genes six-3 and pax-6 are known to induce ectopic lenses during embryogenesis. Here we tested these genes, as well as members of the bone morphogenetic protein (BMP) pathway that regulate establishment of the dorsal-ventral axis in embryos, for their ability to induce lens regeneration. We show that the lens can be regenerated from the ventral iris when the BMP pathway is inhibited and when the iris is transfected with six-3 and treated with retinoic acid. In intact irises, six-3 is expressed at higher levels in the ventral than in the dorsal iris. During regeneration, however, only expression in the dorsal iris is significantly increased. Such an increase is seen in ventral irises only when they are induced to transdifferentiate by six-3 and retinoic acid or by BMP inhibitors. These data suggest that lens regeneration can be achieved in noncompetent adult tissues and that this regeneration occurs through a gene regulatory mechanism that is more complex than the dorsal expression of lens regeneration-specific genes. PMID- 16341015 TI - A colonization factor links Vibrio cholerae environmental survival and human infection. AB - Many bacteria that cause diseases must be able to survive inside and outside the host. Attachment to and colonization of abiotic or biotic surfaces is a common mechanism by which various microorganisms enhance their ability to survive in diverse environments. Vibrio cholerae is a Gram-negative aquatic bacillus that is often found in the environment attached to the chitinous exoskeletons of zooplankton. It has been suggested that attachment to zooplankton enhances environmental survival of Vibrio spp., probably by providing both an abundant source of carbon and nitrogen and protection from numerous environmental challenges. On ingestion by humans, some serogroups of V. cholerae cause the diarrhoeal disease cholera. The pathophysiology of cholera is a result of the effects of cholera toxin on intestinal epithelial cells. For sufficient quantities of cholera toxin to reach the intestinal epithelium and to produce clinical symptoms, colonization of the small bowel must occur. Because most V. cholerae do not colonize humans, but all probably require strategies for survival in the environment, we considered that colonization factors selected for in the environment may be the same as those required for intestinal colonization of humans. In support of this hypothesis, here we have identified a single protein required for efficient intestinal colonization that mediates attachment to both zooplankton and human epithelial cells by binding to a sugar present on both surfaces. PMID- 16341016 TI - Casein kinase 1 gamma couples Wnt receptor activation to cytoplasmic signal transduction. AB - Signalling by Wnt proteins (Wingless in Drosophila) has diverse roles during embryonic development and in adults, and is implicated in human diseases, including cancer. LDL-receptor-related proteins 5 and 6 (LRP5 and LRP6; Arrow in Drosophila) are key receptors required for transmission of Wnt/beta-catenin signalling in metazoa. Although the role of these receptors in Wnt signalling is well established, their coupling with the cytoplasmic signalling apparatus remains poorly defined. Using a protein modification screen for regulators of LRP6, we describe the identification of Xenopus Casein kinase 1 gamma (CK1gamma), a membrane-bound member of the CK1 family. Gain-of-function and loss-of-function experiments show that CK1gamma is both necessary and sufficient to transduce LRP6 signalling in vertebrates and Drosophila cells. In Xenopus embryos, CK1gamma is required during anterio-posterior patterning to promote posteriorizing Wnt/beta catenin signalling. CK1gamma is associated with LRP6, which has multiple, modular CK1 phosphorylation sites. Wnt treatment induces the rapid CK1gamma-mediated phosphorylation of these sites within LRP6, which, in turn, promotes the recruitment of the scaffold protein Axin. Our results reveal an evolutionarily conserved mechanism that couples Wnt receptor activation to the cytoplasmic signal transduction apparatus. PMID- 16341017 TI - A dual-kinase mechanism for Wnt co-receptor phosphorylation and activation. AB - Signalling by the Wnt family of secreted lipoproteins has essential functions in development and disease. The canonical Wnt/beta-catenin pathway requires a single span transmembrane receptor, low-density lipoprotein (LDL)-receptor-related protein 6 (LRP6), whose phosphorylation at multiple PPPSP motifs is induced upon stimulation by Wnt and is critical for signal transduction. The kinase responsible for LRP6 phosphorylation has not been identified. Here we provide biochemical and genetic evidence for a 'dual-kinase' mechanism for LRP6 phosphorylation and activation. Glycogen synthase kinase 3 (GSK3), which is known for its inhibitory role in Wnt signalling through the promotion of beta-catenin phosphorylation and degradation, mediates the phosphorylation and activation of LRP6. We show that Wnt induces sequential phosphorylation of LRP6 by GSK3 and casein kinase 1, and this dual phosphorylation promotes the engagement of LRP6 with the scaffolding protein Axin. We show further that a membrane-associated form of GSK3, in contrast with cytosolic GSK3, stimulates Wnt signalling and Xenopus axis duplication. Our results identify two key kinases mediating Wnt co receptor activation, reveal an unexpected and intricate logic of Wnt/beta-catenin signalling, and illustrate GSK3 as a genuine switch that dictates both on and off states of a pivotal regulatory pathway. PMID- 16341018 TI - The importance of sequence diversity in the aggregation and evolution of proteins. AB - Incorrect folding of proteins, leading to aggregation and amyloid formation, is associated with a group of highly debilitating medical conditions including Alzheimer's disease and late-onset diabetes. The issue of how unwanted protein association is normally avoided in a living system is particularly significant in the context of the evolution of multidomain proteins, which account for over 70% of all eukaryotic proteins, where the effective local protein concentration in the vicinity of each domain is very high. Here we describe the aggregation kinetics of multidomain protein constructs of immunoglobulin domains and the ability of different homologous domains to aggregate together. We show that aggregation of these proteins is a specific process and that the efficiency of coaggregation between different domains decreases markedly with decreasing sequence identity. Thus, whereas immunoglobulin domains with more than about 70% identity are highly prone to coaggregation, those with less than 30-40% sequence identity do not detectably interact. A bioinformatics analysis of consecutive homologous domains in large multidomain proteins shows that such domains almost exclusively have sequence identities of less than 40%, in other words below the level at which coaggregation is likely to be efficient. We propose that such low sequence identities could have a crucial and general role in safeguarding proteins against misfolding and aggregation. PMID- 16341020 TI - TGFbeta pathobiology in the eye. AB - Transforming growth factor beta (TGFbeta), a multifunctional growth factor, is one of the most important ligands involved in the regulation of cell behavior in ocular tissues in physiological or pathological processes of development or tissue repair, although various other growth factors are also involved. Increased activity of this ligand may induce unfavorable inflammatory responses and tissue fibrosis. In mammals, three isoforms of TGFbeta, that is, beta1, beta2, and beta3, are known. Although all three TGFbeta isoforms and their receptors are present in ocular tissues, lack of TGFbeta2, but not TGFbeta1 or TGFbeta3, perturbs embryonic morphogenesis of the eyes in mice. Smads2/3 are key signaling molecules downstream of cell surface receptors for TGFbeta or activin. Upon TGF binding to the respective TGF receptor, Smads2/3 are phosphorylated by the receptor kinase at the C-terminus, form a complex with Smad4 and translocate to the nucleus for activation of TGFbeta gene targets. Moreover, mitogen-activated protein kinase, c-Jun N-terminal kinase, and p38 modulate Smad signals directly via Smad linker phosphorylation or indirectly via pathway crosstalk. Smad signals may therefore be a critical threrapeutic target in the treatment of ocular disorders related to fibrosis as in other systemic fibrotic diseases. The present paper reviews recent progress concerning the roles of TGFbeta signaling in the pathology of the eye. PMID- 16341021 TI - Prenatal nicotine exposure alters the responses to subsequent nicotine administration and withdrawal in adolescence: Serotonin receptors and cell signaling. AB - Offspring of women who smoke during pregnancy are themselves more likely to take up smoking in adolescence, effects that are associated with a high rate of depression and increased sensitivity to withdrawal symptoms. To evaluate the biological basis for this relationship, we assessed effects on serotonin (5 hydroxytryptamine, 5HT) receptors and 5HT-mediated cellular responses in rats exposed to nicotine throughout prenatal development and then given nicotine in adolescence (postnatal days PN30-47.5), using regimens that reproduce plasma nicotine levels found in smokers. Evaluations were then made during the period of adolescent nicotine treatment and for up to one month after the end of treatment. Prenatal nicotine exposure, which elicits damage to 5HT projections in the cerebral cortex and striatum, produced sex-selective changes in the expression of 5HT(1A) and 5HT2 receptors, along with induction of adenylyl cyclase (AC), leading to sensitization of heterologous inputs operating through this signaling pathway. Superimposed on these effects, the AC response to 5HT was shifted toward inhibition. By itself, adolescent nicotine administration, which damages the same pathways, produced similar effects on receptors and the 5HT-mediated response, but a smaller overall induction of AC. Animals exposed to prenatal nicotine showed a reduced response to nicotine administered in adolescence, results in keeping with earlier findings of persistent desensitization. Our results indicate that prenatal nicotine exposure alters parameters of 5HT synaptic communication lasting into adolescence and changes the response to nicotine administration and withdrawal in adolescence, actions which may contribute to a subpopulation especially vulnerable to nicotine dependence. PMID- 16341022 TI - Lobar distribution of lesion volumes in late-life depression: the Biomedical Informatics Research Network (BIRN). AB - White matter hyperintense lesions on T2-weighted images are associated with late life depression. Little work has been carried out examining differences in lesion location between elderly individuals with and without depression. In contrast to previous studies examining total brain white matter lesion volume, this study examined lobar differences in white matter lesion volumes derived from brain magnetic resonance imaging. This study examined 49 subjects with a DSM-IV diagnosis of major depression and 50 comparison subjects without depression. All participants were age 60 years or older. White matter lesion volumes were measured in each hemisphere using a semiautomated segmentation process and localized to lobar regions using a lobar atlas created for this sample using the imaging tools provided by the Biomedical Informatics Research Network (BIRN). The lobar lesion volumes were compared against depression status. After controlling for age and hypertension, subjects with depression exhibited significantly greater total white matter lesion volume in both hemispheres and in both frontal lobes than did control subjects. Although a similar trend was observed in the parietal lobes, the difference did not reach a level of statistical significance. Models of the temporal and occipital lobes were not statistically significant. Older individuals with depression have greater white matter disease than healthy controls, predominantly in the frontal lobes. These changes are thought to disrupt neural circuits involved in mood regulation, thus increasing the risk of developing depression. PMID- 16341023 TI - Visuospatial memory deficits emerging during nicotine withdrawal in adolescents with prenatal exposure to active maternal smoking. AB - Active maternal smoking during pregnancy elevates the risk of cognitive deficits and tobacco smoking among offspring. Preclinical work has shown that combined prenatal and adolescent exposure to nicotine produces more pronounced hippocampal changes and greater deficits in cholinergic activity upon nicotine withdrawal than does prenatal or adolescent exposure to nicotine alone. Few prior studies have examined the potential modifying effects of gestational exposure to active maternal smoking on cognitive or brain functional response to tobacco smoking or nicotine withdrawal in adolescents. We examined visuospatial and verbal memory in 35 adolescent tobacco smokers with prenatal exposure to active maternal smoking and 26 adolescent tobacco smokers with no prenatal exposure to maternal smoking who were similar in age, educational attainment, general intelligence, and baseline plasma cotinine. Subjects were studied during ad libitum smoking and after 24 h of abstinence from smoking. A subset of subjects from each group also underwent functional magnetic resonance imaging while performing a visuospatial encoding and recognition task. Adolescent tobacco smokers with prenatal exposure experienced greater nicotine withdrawal-related deficits in immediate and delayed visuospatial memory relative to adolescent smokers with no prenatal exposure. Among adolescent smokers with prenatal exposure, nicotine withdrawal was associated with increased activation of left parahippocampal gyrus during early recognition testing of visuospatial stimuli and increased activation of bilateral hippocampus during delayed recognition testing of visuospatial stimuli. These findings extend prior preclinical work and suggest that, in human adolescent tobacco smokers, prenatal exposure to active maternal smoking is associated with alterations in medial temporal lobe function and concomitant deficits in visuospatial memory. PMID- 16341026 TI - Hood versus mask nebulization in infants with evolving bronchopulmonary dysplasia in the neonatal intensive care unit. AB - OBJECTIVE: To compare infants' discomfort, nursing-time and caregiver preference, and assess the clinical efficiency (as a secondary outcome) of hood versus facemask nebulization in infants with evolving bronchopulmonary dysplasia (BPD) in the neonatal intensive care unit. STUDY DESIGN: A prospective, open, randomized, controlled crossover clinical trial. In total, 10 infants with BPD who were on inhaled beta-agonist bronchodilators and corticosteroids were randomly assigned to receive their nebulized treatments either by a facemask, or by a hood for 2-3 days, and then crossover to receive the same treatments with the other technique for another 2-3 days. Infants' discomfort, nursing-time, caregiver preference and clinical efficiency were compared. RESULTS: At baseline there was no significant clinical difference between the groups. Nurse-time required for administering the hood nebulization (mean+/-s.e.m.: 1.9+/-0.1 min) was significantly shorter than the time for mask nebulization (12.0+/-0.6 min, P<0.0001). Infants' discomfort score was significantly lower (0.1+/-0.04) for hood versus mask nebulization (2.5+/-0.2, P<0.0001). Nurses and parents unequivocally preferred the hood treatment. During both mask and hood nebulization therapies (2-3 days) clinical efficiency was comparable. While both methods caused an immediate (20 min post) clinical improvement, the immediate respiratory assessment change score was significantly greater for the hood versus the mask nebulization (0.62+/-0.27 versus 0.13+/-0.14, P<0.05). CONCLUSIONS: Nebulization of aerosolized medications in infants with evolving BPD by hood was less time-consuming for caregivers and was much better tolerated by the infants while being at least as effective as the conventional facemask nebulization. PMID- 16341025 TI - The anxiogenic drug yohimbine reinstates palatable food seeking in a rat relapse model: a role of CRF1 receptors. AB - The major problem in treating excessive eating is high rates of relapse to maladaptive eating habits during diet treatments; this relapse is often induced by stress or anxiety states. Preclinical studies have not explored this clinical problem. Here, we adapted a reinstatement model (commonly used to study relapse to abused drugs) to examine the role of stress and anxiety in relapse to palatable food seeking during dieting. Rats were placed on restricted diet (75 80% of daily standard food) and for 12 intermittent training days (9 h/day, every other day) lever-pressed for palatable food pellets (25% fat, 48% carbohydrate) under a fixed ratio 1 (20-s timeout) reinforcement schedule. Subsequently, the rats were given 10 daily extinction sessions during which lever presses were not reinforced, and were then injected with yohimbine (an alpha-2 adrenoceptor antagonist that induces stress and anxiety in humans and non-humans) or given a single food pellet to assess reinstatement of food seeking. The rats rapidly learned to lever press for the palatable pellets and across the training days the ratio of timeout nonreinforced lever presses to reinforced lever presses progressively increased more than three-fold, suggesting the development of compulsive eating behavior. After extinction, yohimbine injections and pellet priming reliably reinstated food seeking. The corticotropin-releasing factor1 (CRF1) receptor antagonist antalarmin attenuated the reinstatement induced by yohimbine, but not pellet priming. Antalarmin also reversed yohimbine's anxiogenic effects in the social interaction test. These data suggest that CRF is involved in stress-induced relapse to palatable food seeking, and that CRF1 antagonists should be considered for the treatment of maladaptive eating habits. PMID- 16341024 TI - Activation of group II metabotropic glutamate receptors in the nucleus accumbens shell attenuates context-induced relapse to heroin seeking. AB - Using a rat relapse model, we previously reported that re-exposing rats to a drug associated context, following extinction of operant responding in a different context, reinstates heroin seeking. In an initial pharmacological characterization, we found that the mGluR2/3 agonist LY379268, which acts centrally to reduce evoked glutamate release, attenuates context-induced reinstatement of heroin seeking when injected systemically or into the ventral tegmental area, the cell body region of the mesolimbic dopamine system. Here, we tested whether injections of LY379268 into the nucleus accumbens (NAc), a terminal region of the mesolimbic dopamine system, would also attenuate context induced reinstatement of heroin seeking. Rats were trained to self-administer heroin; drug infusions were paired with a discrete tone-light cue. Subsequently, lever pressing was extinguished in the presence of the discrete cue in a context that differed from the drug self-administration context in terms of visual, auditory, tactile, and circadian cues. After extinction of responding, LY379268 was injected to different groups of rats into the NAc core or shell or into the caudate-putamen, a terminal region of the nigrastriatal dopamine system. Injections of LY379268 into the NAc shell (0.3 or 1.0 microg) dose-dependently attenuated context-induced reinstatement of heroin seeking. Injections of 1.0 microg of LY379268 into the NAc core had no effect, while a higher dose (3.0 microg) decreased this reinstatement. Injections of LY379268 (3.0 microg) 1.5 mm dorsal from the NAc core into the caudate-putamen were ineffective. Results suggest an important role of glutamate transmission in the NAc shell in context induced reinstatement of heroin seeking. PMID- 16341027 TI - Sustained reductions in neonatal nosocomial infection rates following a comprehensive infection control intervention. AB - OBJECTIVE: Nosocomial infections (NI) are a frequent and important cause of morbidity and mortality in newborn infants who receive intensive care. We sought to determine if comprehensive infection control (CIC) measures decrease rates in a large neonatal intensive care nursery. METHODS: Single center interventional study. The CIC intervention consisted of increasing nursing and physician education and awareness of infection rates, establishing common improvement goals, training in hand and environment care, and implementing a specialty nursing team for central venous and arterial catheter care. Demographic and microbiology information for all infants admitted to the NICU from January 1, 1999 to December 31, 2000 established baseline data. The intervention period was during January and February 2001. The postintervention period was March 1, 2001 to February 29, 2004. The main outcome measure was the rate of blood, cerebrospinal and/or urinary tract bacterial infections per 1000 hospital days. RESULTS: Baseline infection rate was 8.5 per 1000 hospital days. The NI rate fell 26% (P=0.002) from baseline in the first year and 29% (P<0.001) in the second and third years after the CIC intervention. The reduction in total NI was due mostly to a 46% fall in coagulase-negative Staphylococcus infection rate (P<0.001); however, rates of all other organisms also fell by 21% (P=0.05). CONCLUSIONS: CIC measures can reduce bacterial and fungal NI rates. This effect has been sustained for 3 years following the intervention. PMID- 16341028 TI - Injection of skeletal muscle-derived cells into the penis improves erectile function. AB - We investigated the effect of intrapenile injection of muscle-derived cells (MDC) on the erectile function in rats with bilateral cavernous nerve injury. Rat MDC were harvested and transduced with a retrovirus expressing the lacZ gene. Hanks' balanced salt solution (HBSS) (20 microl) or MDC (1 x 10(6) cells/side) were injected in each corpora cavernosa immediately before bilateral cavernous nerve transection. Intracavernous pressures (ICP) were measured 2 or 4 weeks after surgery with electrical stimulation of the pelvic nerves. Mean maximal ICP of sham group was significantly lower than that of control group both at 2 and 4 weeks after surgery. When MDC were injected into the penis, ICP improved over the sham-injected group at both 2 and 4 weeks after surgery. Percent area of PGP 9.5 staining was significantly greater in MDC-injected penis than in sham-injected at 2 and 4 weeks. Penile MDC injection can facilitate recovery of injured penile innervation and improve erectile function. PMID- 16341029 TI - A comparison between portable ultrasound (MIDUS) and nocturnal RigiScan when confirming the diagnosis of vascular organic erectile disorder. AB - In recent years, the use of RigiScan and ultrasound to assess erectile dysfunction has fallen from favour. However, in a small minority of specialist cases, where a vascular, neurogenic or psychogenic aetiology requires confirmation, there remains a need for further investigation. To establish if in a preliminary assessment the use of nocturnal RigiScan or male impotence diagnostic ultrasound system (MIDUS) represents best practice as a diagnostic investigation in patients with a history suggestive of vascular organic erectile disorder. Men attending both urological and psychosexual therapy clinics with erectile dysfunction were assessed using a generic assessment schedule. Patients with a history suggestive of vascular erectile disorder were offered the opportunity of dual investigation of their condition. After screening using a provocative RigiScan using visual stimuli that gleaned inconclusive results, patients were offered the chance to enter a study with both nocturnal RigiScan and MIDUS investigation. These were confined for the purposes of this study to RigiScan events, peak systolic flow velocity (PSV) and end-diastolic flow velocity (EDV) from ultrasound examination where an abnormal EDV is defined as in excess of 4.5 cm/s and a normal PSV is variously defined as being greater than 35 cm/s. In all, 38/43 (88%, 95% CI: 76-95%) of men had a nocturnal event exceeding 3 min on the RigiScan investigation. This compares with 17/43 (40%, 95% CI: 26 54%) of men with a normal EDV blood flow of less than 4.5 cm/s (P<0.017) and 32/43 (74%, CI: 60-85%) of men with a normal PSV flow greater than 35 cm/s (NS). Rigiscan and ultrasonography of the cavernosal vessels are of equal usefulness in suspected arterial penile disease although where veno-occlusive disease is suspected, ultrasonography is more specific. PMID- 16341030 TI - Early work on apoptosis, an interview with Sten Orrenius. PMID- 16341031 TI - The p53 family inhibitor DeltaNp73 interferes with multiple developmental programs. PMID- 16341032 TI - Detection of JAK2 V617F as a first intention diagnostic test for erythrocytosis. PMID- 16341033 TI - Aberrant subcellular expression of nucleophosmin and NPM-MLF1 fusion protein in acute myeloid leukaemia carrying t(3;5): a comparison with NPMc+ AML. PMID- 16341034 TI - Chronic myeloproliferative disorders: a tyrosine kinase tale. AB - Chronic myeloproliferative diseases (CMPDs) are characterized by the abnormal proliferation and survival of one or more myeloid cell types. The archetype of this class of hematological diseases is chronic myeloid leukemia (CML), characterized by the presence of the Philadelphia (Ph) chromosome, the result of t(9;22)(q34;q11), and the associated BCR-ABL1 oncogene. Some of the Ph-negative myeloproliferative diseases are characterized by other chromosomal translocations involving a variety of tyrosine kinase genes, including ABL1, ABL2, PDGFRA, PDGFRB, FGFR1, and JAK2. The majority of Ph-negative CMPDs, however, such as chronic eosinophilic leukemia, polycythemia vera, essential thrombocythemia, and idiopathic myelofibrosis are not characterized by the presence of recurrent chromosomal abnormalities. Recent studies have identified the FIP1L1-PDGFRA fusion gene, generated due to a small cryptic deletion on chromosome 4q12, and the activating V617F mutation in JAK2 in a significant fraction of Ph-negative CMPDs. These results show that abnormalities in tyrosine kinase genes are central to the molecular pathogenesis of CMPDs. Genome-wide screenings to identify novel tyrosine kinase abnormalities in CMPDs may contribute to further improvement of the diagnosis and the treatment of these diseases. PMID- 16341035 TI - Loss of the NPM1 gene in myeloid disorders with chromosome 5 rearrangements. AB - The assignment with chromosome banding techniques of the breakpoints of the recurrent translocation t(3;5) which leads to NPM1/MLF1 gene fusion in myeloid malignancies has not been unequivocal. In order to assess whether this is due to uncertainty in interpretation of the observed banding pattern or whether it reflects true genomic heterogeneity, we decided to analyze the breakpoint positions using fluorescence in situ (FISH) techniques in eight patients with myeloid malignancies and rearrangements of chromosomes 3 and 5. In three patients, colocalization of the NPM1 and MLF1 spanning BACs was demonstrated and NPM1/MLF1 fusion shown by PCR in one while in the remaining cases breakpoints were located outside the NPM1 and MLF1 loci. Interestingly, loss of a copy of the NPM1 gene was found in three of these latter patients. This findings suggest that haploinsufficiency of NPM1 may play a role in subtypes of myelodysplasias and leukemias. PMID- 16341036 TI - A novel t(3;17)(p25;q21) variant translocation of acute promyelocytic leukemia with rearrangement of the RARA locus. PMID- 16341037 TI - Immunomodulatory effects of Toll-like receptor-7 activation on chronic lymphocytic leukemia cells. AB - Weak immunogenicity of chronic lymphocytic leukemia (CLL) cells may contribute to disease progression and inhibit effective immunotherapy. Accordingly, agents that enhance the immunogenicity of CLL cells may be useful in immunotherapeutic approaches to this disease. Since Toll-like receptors (TLRs) are major regulators of innate immunity and initiation of adaptive immunity, we studied the effects of viral pathogen associated molecular pattern agonists (that are recognized by TLRs) on the costimulatory phenotype and function of CLL cells. CLL cells (especially those with high endogenous expression of CD38) responded to TLR7 activating imidazoquinolines and guanosine analogs by increasing costimulatory molecule expression, producing inflammatory cytokines, and becoming more sensitive to killing by cytotoxic effectors. Additional activation of protein kinase C pathways increased the ability to stimulate T-cell proliferation, blocked phosphorylation of the transcription factor, signal transducer and activator of transcription (STAT)3, and resulted in the acquisition of a dendritic cell surface phenotype by TLR7-activated CLL cells. Normal B cells also responded to TLR7 activation by increasing costimulatory molecule expression and cytokine production. These findings suggest a potential role for TLR7 agonists in CLL immunotherapy. PMID- 16341038 TI - Mutational screen reveals a novel JAK2 mutation, L611S, in a child with acute lymphoblastic leukemia. PMID- 16341039 TI - Genome-wide approach to identify risk factors for therapy-related myeloid leukemia. AB - Using a target gene approach, only a few host genetic risk factors for treatment related myeloid leukemia (t-ML) have been defined. Gene expression microarrays allow for a more genome-wide approach to assess possible genetic risk factors for t-ML. We assessed gene expression profiles (n=12 625 probe sets) in diagnostic acute lymphoblastic leukemic cells from 228 children treated on protocols that included leukemogenic agents such as etoposide, 13 of whom developed t-ML. Expression of 68 probes, corresponding to 63 genes, was significantly related to risk of t-ML. Hierarchical clustering of these probe sets clustered patients into three groups with 94, 122 and 12 patients, respectively; 12 of the 13 patients who went on to develop t-ML were overrepresented in the latter group (P<0.0001). A permutation test indicated a low likelihood that these probe sets and clusters were obtained by chance (P<0.001). Distinguishing genes included transcription related oncogenes (v-Myb, Pax-5), cyclins (CCNG1, CCNG2 and CCND1) and histone HIST1H4C. Common transcription factor recognition elements among similarly up- or downregulated genes included several involved in hematopoietic differentiation or leukemogenesis (Maz, PU.1, ARNT). This approach has identified several genes whose expression distinguishes patients at risk of t-ML, and suggests targets for assessing germline predisposition to leukemogenesis. PMID- 16341040 TI - Frequent elevation of Akt kinase phosphorylation in blood marrow and peripheral blood mononuclear cells from high-risk myelodysplastic syndrome patients. AB - The serine/threonine kinase Akt, a downstream effector of phosphatidylinositol 3 kinase (PI3K), is known to play an important role in antiapoptotic signaling and has been implicated in the aggressiveness of a number of different human cancers including acute myeloid leukemia (AML). The progression of myelodysplastic syndromes (MDSs) to AML is thought to be associated with abrogation of apoptotic control mechanisms. However, little is known about signal transduction pathways which may be involved in enhanced survival of MDS cells. In this report, we have performed immunocytochemical and flow cytometric analysis to evaluate the levels of activated Akt in bone marrow or peripheral blood mononuclear cells from patients diagnosed with MDS. We observed high levels of Ser473 phosphorylated Akt (p-Akt) staining in 90% of the cases (n=22) diagnosed as high-risk MDS, whereas mononuclear cells from normal bone marrow or low-risk MDS patients showed low or absent Ser473 p-Akt staining. Furthermore, all high-risk MDS patients also demonstrated high expression of the Class I PI3K p110delta catalytic subunit and a decreased expression of PTEN. Taken together, our results suggest that Akt activation might be one of the factors contributing to the decreased apoptosis rate observed in patients with high-risk MDS. PMID- 16341041 TI - Constitutive phosphoinositide 3-kinase activation in acute myeloid leukemia is not due to p110delta mutations. PMID- 16341042 TI - Imatinib mesylate is effective in children with chronic myelogenous leukemia in late chronic and advanced phase and in relapse after stem cell transplantation. AB - A multicentric phase 2 study was conducted to determine the efficiency and the tolerance of imatinib mesylate in children with chronic myelogenous leukemia (CML) in advanced phase of the disease, in relapse after stem cell transplantation, or in case of failure to an interferon alpha-based regimen. In all, 30 children from eight European countries were enrolled. In 18 children assessable for hematologic response, imatinib mesylate induced complete hematologic response in eight (80%) of the 10 patients included in chronic phase and in six (75%) of eight enrolled in advanced phase of the disease with acceptable toxicity. In 27 patients assessable for cytogenetic response, imatinib mesylate induced disappearance of Philadelphia chromosome-positive bone marrow cells in 12 (60%) of 20 children included in chronic phase and in two (29%) of seven included in advanced phase. A reduction of the bcr-abl/abl ratio to less than 10(-4) was achieved in 11 (50%) of the children included in chronic phase. Estimated 12-month overall survival rate was 95% (95% CI, 87-100%) for the patients included in chronic phase and 75% (95%CI, 45-100%) for those enrolled in advanced phase. Imatinib mesylate is well tolerated and molecular remission can be achieved in children with CML. PMID- 16341043 TI - Wilms' tumor gene 1 (WT1) expression in childhood acute lymphoblastic leukemia: a wide range of WT1 expression levels, its impact on prognosis and minimal residual disease monitoring. AB - Wilms' tumor gene 1 (WT1) is overexpressed in the majority (70-90%) of acute leukemias and has been identified as an independent adverse prognostic factor, a convenient minimal residual disease (MRD) marker and potential therapeutic target in acute leukemia. We examined WT1 expression patterns in childhood acute lymphoblastic leukemia (ALL), where its clinical implication remains unclear. Using a real-time quantitative PCR designed according to Europe Against Cancer Program recommendations, we evaluated WT1 expression in 125 consecutively enrolled patients with childhood ALL (106 BCP-ALL, 19 T-ALL) and compared it with physiologic WT1 expression in normal and regenerating bone marrow (BM). In childhood B-cell precursor (BCP)-ALL, we detected a wide range of WT1 levels (5 logs) with a median WT1 expression close to that of normal BM. WT1 expression in childhood T-ALL was significantly higher than in BCP-ALL (P<0.001). Patients with MLL-AF4 translocation showed high WT1 overexpression (P<0.01) compared to patients with other or no chromosomal aberrations. Older children (> or =10 years) expressed higher WT1 levels than children under 10 years of age (P<0.001), while there was no difference in WT1 expression in patients with peripheral blood leukocyte count (WBC) > or =50 x 10(9)/l and lower. Analysis of relapsed cases (14/125) indicated that an abnormal increase or decrease in WT1 expression was associated with a significantly increased risk of relapse (P=0.0006), and this prognostic impact of WT1 was independent of other main risk factors (P=0.0012). In summary, our study suggests that WT1 expression in childhood ALL is very variable and much lower than in AML or adult ALL. WT1, thus, will not be a useful marker for MRD detection in childhood ALL, however, it does represent a potential independent risk factor in childhood ALL. Interestingly, a proportion of childhood ALL patients express WT1 at levels below the normal physiological BM WT1 expression, and this reduced WT1 expression appears to be associated with a higher risk of relapse. PMID- 16341044 TI - Novel t(5;9)(q33;q22) fuses ITK to SYK in unspecified peripheral T-cell lymphoma. AB - Among peripheral T-cell lymphomas (PTCL), the heterogeneous category of unspecified PTCL represents the most common subtype. Nevertheless, recurrent chromosomal translocations are unknown in this aggressive type of lymphoma. Here we describe a novel t(5;9)(q33;q22) in unspecified PTCL. Molecular analyses delineated the breakpoints to ITK and SYK resulting in a previously undescribed expression of the Syk tyrosine kinase by Itk. ITK-SYK transcripts were detected in five of 30 (17%) unspecified PTCL, but not in cases of angioimmunoblastic T cell lymphoma (n=9) and anaplastic lymphoma kinase-negative anaplastic large-cell lymphoma (n=7). In all five translocation-positive cases, the breakpoints were identical fusing the N-terminal pleckstrin homology domain and proline-rich region of ITK to the tyrosine kinase domain of SYK. Three of the five t(5;9)(q33;q22)+ unspecified PTCL shared a very similar histological pattern with predominant involvement of lymphoid follicles and the same CD3+CD5+CD4+bcl 6+CD10+ immunophenotype. These results demonstrate the presence of a recurrent t(5;9)(q33;q22) in a subset of unspecified PTCL, which may represent a novel distinct subgroup of PTCL. PMID- 16341046 TI - Characterization of fusion partner genes in 114 patients with de novo acute myeloid leukemia and MLL rearrangement. AB - The fusion transcripts of MLL rearrangement [MLL(+)] in acute myeloid leukemia (AML) and their clinicohematologic correlation have not be well characterized in the previous studies. We used Southern blot analysis to screen MLL(+) in de novo AML. Reverse transcriptase-polymerase chain reaction was used to detect the common MLL fusion transcripts. cDNA panhandle PCR was used to identify infrequent or unknown MLL partner genes. MLL(+) was identified in 114 (98 adults) of 988 AML patients. MLL fusion transcripts comprised of 63 partial tandem duplication of MLL (MLL-PTD), 14 MLL-AF9, 9 MLL-AF10, 9 MLL-ELL, 8 MLL-AF6, 4 MLL-ENL and one each of MLL-AF1, MLL-AF4, MLL-MSF, MLL-LCX, MLL-LARG, MLL-SEPT6 and MLL-CBL. The frequency of MLL-PTD was 7.1% in adults and 0.9% in children (P<0.001). 11q23 abnormalities were detected in 64% of MLL/t11q23 and in none of MLL-PTD by conventional cytogenetics. There were no differences in remission rate, event free survival and overall survival between adult MLL-PTD and MLL/t11q23 groups. Adult patients had a significantly poorer outcome than children. The present study showed that cDNA panhandle PCR can identify all rare or novel MLL partner genes. MLL-PTD was rare in childhood AML. MLL(+) adults had a poor outcome with no difference in survival between MLL-PTD and MLL/t11q23 groups. PMID- 16341047 TI - Engraftment capacity of mesenchymal cells following hematopoietic stem cell transplantation in patients receiving reduced-intensity conditioning regimen. AB - The engraftment ability of mesenchymal cells was investigated in 26 patients receiving allogeneic transplantation from HLA-identical siblings with reduced intensity conditioning (RIC). The stem cell source was bone marrow (BM) in eight patients and G-CSF-mobilized peripheral blood hematopoietic cells in 18 cases. A total of 32 patients engrafted very quickly and the chimerism evaluation (both on myeloid and on lymphoid subsets) showed that they were full donor by day 60. At the time of the study they were in complete hematological remission and displayed a full donor hematopoiesis. Two patients showed early disease progression while one did not engraft. Forty-eight out-marrow samples harvested from the 26 patients generated a marrow stromal layer adequate for the chimerism evaluation. Monocyte-macrophage contamination of marrow stromal layers was always reduced below 2% by repeated trypsinizations and treatment with the leucyl-leucine (leu leu) methyl ester. The chimerism evaluation was performed by PCR analysis of STRs microsatellites and the amelogenin locus, by using capillary electrophoresis (CE) and by FISH analysis in case of the sex mismatch. In eight patients, a partial donor origin of stromal cells was shown (7-86% cells of donor). The source of hematopoietic cells was BM in three patients and mobilized peripheral blood in the other five. PMID- 16341048 TI - TCL1 shows a regulated expression pattern in chronic lymphocytic leukemia that correlates with molecular subtypes and proliferative state. AB - Expression of the human oncogene TCL1 in transgenic mice produces B-cell tumors that resemble chronic lymphocytic leukemia (CLL) suggesting its role in B-cell tumorigenesis. To clarify the expression pattern and regulation of TCL1 in CLL, we assessed 213 primary tumors by immunohistochemistry (IHC), flow-cytometry and/or Western blot, using a new monoclonal antibody. TCL1 protein was detectable in the majority of CLL (90% by IHC) but showed marked variations across cases with virtual absence in approximately 10% of tumors. Higher TCL1 levels correlated with markers of the 'pre-germinal center' CLL subtype including unmutated VH status (P=0.005), ZAP70 expression (P=0.007), and presence of chromosome 11q22-23 deletions (P=0.04). Intratumoral heterogeneity in TCL1 levels was also prominent and explained in part by markedly lower TCL1 expression in proliferating tumor cells. In vitro exposure of CLL cells to interleukin-4 (but not other growth factors) produced progressive and irreversible decrease in TCL1 protein levels in association with the onset of proliferation. TCL1 expression patterns in CLL are complex and highly dynamic and appear to reflect both the histogenetic subtypes of the disease and the growth parameters of individual tumors. The observed regulation pattern suggests that TCL1 may exert its effects predominantly in the unmutated/ZAP70-positive tumor subset. PMID- 16341049 TI - Alemtuzumab induces caspase-independent cell death in human chronic lymphocytic leukemia cells through a lipid raft-dependent mechanism. AB - Alemtuzumab is a humanized IgG1 kappa antibody directed against CD52, a glycosyl phosphatidylinositol linked cell-membrane protein of unknown function. Herein, we demonstrate that alemtuzumab promotes rapid death of chronic lymphocytic leukemia (CLL) cells in vitro, in a complement and accessory cell free system. Using minimal detergent solubilization of CLL membranes, we found that CD52 colocalizes with ganglioside GM-1, a marker of membrane rafts. Fluorescence microscopy revealed that upon crosslinking CD52 with alemtuzumab+anti-Fc IgG, large patches, and in many cases caps, enriched in CD52 and GM-1 formed upon the CLL cell plasma membrane. Depletion of membrane cholesterol or inhibition of actin polymerization significantly diminished the formation of alemtuzumab-induced caps and reduced alemtuzumab-mediated CLL cell death. We compared alemtuzumab-induced direct cytotoxicity, effector cell-mediated toxicity and complement-mediated cytotoxicity of CLL cells to normal T cells. The direct cytotoxicity and observed capping was significantly greater for CLL cells as compared to normal T cells. Cell-mediated and complement-mediated cytotoxicity did not significantly differ between the two cell types. In summary, our data support the hypothesis that alemtuzumab can initiate CLL cell death by crosslinking CD52-enriched lipid rafts. Furthermore, the differential direct cytotoxic effect suggests that CD52 directed antibodies could possibly be engineered to more specifically target CLL cells. PMID- 16341050 TI - Identification of circulating CD10 positive T cells in angioimmunoblastic T-cell lymphoma. AB - In most cases of lymphomas with blood dissemination, the careful cytological analysis of peripheral blood smears provides a rapid orientation to diagnosis, even if the final subtyping is achieved by histology and eventually other techniques. Here, we evaluated if the analysis of blood smears may suggest the blood dissemination of angioimmunoblastic T-cell lymphoma (AITL) and if CD10 expression on neoplastic T cells, as recently reported on AITL, may contribute to the diagnosis. In all, 11 lymph nodes and six peripheral blood samples from 12 patients with AITL were studied using four-colour flow cytometry associated to histological, cytological and molecular data. According to previous results, a fraction of T cells expressed CD10 in 10/11 lymph nodes. Interestingly, all blood smears showed atypical lymphoid cells and a fraction of T cells expressed CD10 with a mean percentage of 18.75% (range 5.00-47.00%), regardless of lymphocytosis level and of rate of CD10 T cells in corresponding lymph node. In contrast, in all control samples (100), none CD10-positive T cell was identified. This is to our knowledge the first description of circulating CD10 neoplastic T cells in AITL. Therefore, they ought to be explored in further studies when aggressive lymphoma, in particular with lymphopenia and circulating atypical cells, is suspected. PMID- 16341051 TI - Renin-angiotensin system polymorphisms interact with blood pressure in the determination of microalbuminuria in the general population. PMID- 16341052 TI - Evaluation of health-related quality of life among hypertensive patients in a rural area, PR China. PMID- 16341053 TI - Byproducts of oxidative protein damage and antioxidant enzyme activities in plasma of patients with different degrees of essential hypertension. AB - Despite evidence that essential hypertension (EH) is a state of increased oxidative stress, the data on oxidative protein modifications is lacking. Besides, the role of extracellular antioxidant enzymes in EH has not been systematically studied. Study was performed in 45 subjects with EH and 25 normotensive controls. Patients were divided into three groups according to the 2003 ESH/ESC guidelines (grade 1-3). Plasma protein reactive carbonyl derivatives (RCD) and SH-groups (as byproducts of oxidative protein damage) as well as antioxidant enzyme activities superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase were studied spectrophotometrically and correlated with blood pressure (BP). RCD levels were increased in EH patients compared to controls and correlated significantly with both systolic blood pressure (SBP) (r = 0.495, P<0.01) and diastolic blood pressure (DBP) (r = 0.534, P<0.01). Plasma SH-groups content was significantly lower in all patients with EH, with no correlation with BP. SOD and catalase activity in patients with grade 1 EH were similar to that of controls. Patients with grade 2 and 3 of EH had lower SOD and catalase activity. However, significant correlation with SBP and DBP was observed for catalase only (r = -0.331; P<0.05 and r = -0.365; P<0.05, respectively). EH patients exhibited higher plasma GPX activity compared to those in controls, and it correlated with SBP (r = 0.328; P<0.05). The results presented show that increased oxidative protein damage is present in all grades of EH. In mild hypertension extracellular antioxidant enzyme activities are not decreased, suggesting they are probably not critical in early EH, but could be important in moderate to severe EH. PMID- 16341054 TI - An investigation of transmission ratio distortion in the central region of the human MHC. AB - Transmission ratio distortion (TRD) describes a significant departure from expected Mendelian inheritance ratios that is fundamental to both the biology of reproduction and statistical genetics. The relatively high fetal wastage in humans, with consequent selection of alleles in utero, makes it likely that TRD is prevalent in the human genome. The central region of the human major histocompatibility complex (MHC) is a strong TRD candidate, as it houses a number of immune and regulatory genes that may be important in pregnancy outcome. We used a nonhaplotype-based method to select 13 tagging SNPs from three central MHC candidate regions, and analysed their transmission in 380 newborns and their parents (1138 individuals). A TRD of 54:46 was noted in favour of the common allele of a promoter SNP in the CLIC1 gene (P = 0.025), with a similar distortion using haplotypes across the same gene region (P = 0.016). We also found evidence that markers in the CLIC1 gene region may have been subject to recent selection (P < 0.001). The study illustrates the potential benefits of screening for TRD and highlights the difficulties encountered therein. PMID- 16341055 TI - Examination of seven candidate regions for multiple sclerosis: strong evidence of linkage to chromosome 1q44. AB - Multiple sclerosis (MS) is a debilitating neuroimmunological and neurodegenerative disease with a strong genetic component. Numerous studies have failed to consistently identify genes that confer disease susceptibility except for association with HLA-DR. Seven non-HLA regions (1q, 2q, 9q, 13q, 16q, 18p and 19q) identified in a recent genomic screen were investigated by genotyping approximately 20 single-nucleotide polymorphisms (SNPs) at approximately 1 Mb intervals. Non-parametric multipoint analyses identified a peak LOD* score of 2.99 for the 1q44 region and substantially narrowed the linkage peak to approximately 7 Mb. Ordered subset analyses (OSA) identified significant LOD score increases for 2q35 and 18p11 when ranking families by HLA-DR status and identified a significant LOD score increase in region 2q35 when ranking families by linkage to chromosome 1q44. 1q44 is particularly interesting because of linkage evidence for this region in studies of both rheumatoid arthritis and systemic lupus erythematosus. PMID- 16341056 TI - Genospheres: self-assembling nucleic acid-lipid nanoparticles suitable for targeted gene delivery. AB - We describe the assembly of a cationic lipid-nucleic acid nanoparticle from a liquid monophase containing water and a water miscible organic solvent where both lipid and DNA components are separately soluble prior to their combination. Upon removal of the organic solvent, stable and homogenously sized (70-100 nm) lipid nucleic acid nanoparticles (Genospheres) were formed. The low accessibility (<15%) of the nanoparticle-encapsulated DNA to a DNA intercalating dye indicated well-protected nucleic acids and high DNA incorporation efficiencies. It was demonstrated that Genospheres could be stably stored under a variety of conditions including a lyophilized state where no appreciable increase in particle size or DNA accessibility was observed following reconstitution. Finally, Genospheres were made target-specific by insertion of an antibody lipopolymer (anti-HER2 scFv (F5)-PEG-DSPE) conjugate into the particle. The target specificity (>100-fold) in HER2 overexpressing SK-BR-3 breast cancer cells was dependent on the degree of PEGylation, where the incorporation of high amounts of PEG-lipid on the particle surface (up to 5 mol%) had only a minor effect on the transfection activity of the targeted Genospheres. In summary, this work describes a novel, readily scalable method for preparing highly stable immunotargeted nucleic acid delivery vehicles capable of achieving a high degree of specific transfection activity. PMID- 16341057 TI - Ring-Sp1 decoy oligonucleotide effectively suppresses extracellular matrix gene expression and fibrosis of rat kidney induced by unilateral ureteral obstruction. AB - Tubulointerstitial fibrosis is the consequence of an injury characterized by the accumulation of excess collagen and other extracellular matrix components, resulting in the destruction of the normal kidney architecture and subsequent loss of function. A transcription factor Sp1, originally described as a ubiquitous transcription factor, is involved in the basal expression of extracellular matrix genes and may, therefore, be important in fibrotic processes. Here, we report on the design of a ring-Sp1 decoy oligonucleotide, containing the consensus Sp1 binding sequence in a single decoy molecule without an open end, to create a novel therapeutic strategy for fibrosis. The ring-Sp1 decoy oligonucleotide is highly resistant to degradation by nucleases or serum compared to the conventional phosphorothioated double-stranded Sp1 decoy oligonucleotide, and effectively suppressed the expression of transforming growth factor-beta1 and fibronectin, the binding of Sp1 to the promoter region of these genes, and proliferation in response to serum in normal rat kidney fibroblasts. Moreover, treatment with the ring-Sp1 decoy in vivo significantly attenuates extracellular matrix gene expression in the rat kidney in which a unilateral ureteral obstruction had been induced. These results suggest that the ring-Sp1 decoy oligonucleotide represents promising therapeutic alternative to the conventional treatment of fibrotic disorders. PMID- 16341058 TI - Inhibition of CD26 peptidase activity significantly improves engraftment of retrovirally transduced hematopoietic progenitors. AB - It has previously been shown that inhibition of CD26 (DPPIV/dipeptidylpeptidase IV) peptidase activity improves homing of hematopoietic stem cells (HSCs) to the bone marrow and increases engraftment efficiency. Here, we demonstrate that treatment of retrovirally transduced mouse bone marrow cells with the tri-peptide Diprotin A (Ile-Pro-Ile), a specific inhibitor of CD26, significantly enhances engraftment of retrovirally transduced HSCs. Treatment of transduced bone marrow cells with Diprotin A permitted long-term expression of a retrovirally encoded MHC class I gene on multiple hematopoietic cell lineages after transplantation of a suboptimal number of transduced cells. Secondary transfer experiments revealed that expression of the transduced MHC class I gene resulted from engraftment of transduced HSCs. Expression of the allogeneic MHC class I antigen on bone marrow derived cells following transplantation of Diprotin A-treated cells was sufficient to induce transplantation tolerance. Therefore, inhibition of CD26 activity significantly enhances engraftment of limited numbers of genetically modified HSCs, resulting in physiologically relevant levels of gene transfer. PMID- 16341059 TI - Prospects of RNA interference therapy for cancer. AB - RNA interference (RNAi) is a powerful gene-silencing process that holds great promise in the field of cancer therapy. The discovery of RNAi has generated enthusiasm within the scientific community, not only because it has been used to rapidly identify key molecules involved in many disease processes including cancer, but also because RNAi has the potential to be translated into a technology with major therapeutic applications. Our evolving understanding of the molecular pathways important for carcinogenesis has created opportunities for cancer therapy employing RNAi technology to target the key molecules within these pathways. Many gene products involved in carcinogenesis have already been explored as targets for RNAi intervention, and RNAi targeting of molecules crucial for tumor-host interactions and tumor resistance to chemo- or radiotherapy has also been investigated. In most of these studies, the silencing of critical gene products by RNAi technology has generated significant antiproliferative and/or proapoptotic effects in cell-culture systems or in preclinical animal models. Nevertheless, significant obstacles, such as in vivo delivery, incomplete suppression of target genes, nonspecific immune responses and the so-called off-target effects, need to be overcome before this technology can be successfully translated into the clinical arena. Significant progress has already been made in addressing some of these issues, and it is foreseen that early phase clinical trials will be initiated in the very near future. PMID- 16341060 TI - Enhanced transduction of mouse salivary glands with AAV5-based vectors. AB - We previously demonstrated that recombinant adeno-associated virus vectors based on serotype 2 (rAAV2) can direct transgene expression in salivary gland cells in vitro and in vivo. However, it is not known how other rAAV serotypes perform when infused into salivary glands. The capsids of serotypes 4 and 5 are distinct from rAAV2 and from each other, suggesting that they may direct binding and entry into different cell types. In the present study, we investigated the tropisms, transduction efficiencies, and antibody response to AAV vectors based on AAV serotypes 2, 4, and 5. Administration of rAAV2beta-galactosidase (betagal), rAAV4betagal, or rAAV5betagal to murine submandibular salivary glands by retrograde ductal instillation resulted in efficient transduction of salivary epithelial cells, with AAV4 and AAV5 producing 2.3 and 7.3 times more betagal activity compared with AAV2. Improved transduction with AAV5 was confirmed by QPCR of DNA extracted from glands and immunohistochemical staining for transgene expression. Like AAV2, AAV5 primarily transduced striated and intercalated ductal cells. AAV4 transduction was evident in striated, intercalated, and excretory ductal cells, as well as in convoluted granular tubules. In keeping with the encapsulated nature of the salivary gland, the majority of persistent viral genomes were found in the gland and not in other tissues. Neutralizing antibodies (NABs) found in the serum of virus-infused animals were serotype specific and there was no crossreactivity between serotypes. No NABs were detected in saliva but sialic acid conjugates present in saliva could neutralize AAV4 at low dilutions. Together our data suggest that because of differences in receptor binding and transduction pathways, other serotypes may have improved utility as gene transfer vectors in the salivary gland and these differences could be exploited in gene therapy applications. PMID- 16341061 TI - Innovation: Rescuing drug discovery: in vivo systems pathology and systems pharmacology. AB - The pharmaceutical industry is currently beleaguered by close scrutiny from the financial community, regulators and the general public. Productivity, in terms of new drug approvals, has generally been falling for almost a decade and the safety of a number of highly successful drugs has recently been brought into question. Here, we discuss whether taking an in vivo systems approach to drug discovery and development could be the paradigm shift that rescues the industry. PMID- 16341062 TI - Opinion: NK3 receptor antagonists: the next generation of antipsychotics? AB - Although current antipsychotic drugs are effective at treating the psychotic (positive) symptoms of schizophrenia, they have one or more serious side effects, including extrapyramidal symptoms, weight gain, cardiovascular liabilities and type II diabetes. However, recent data from clinical trials of selective neurokinin 3 (NK(3)) receptor antagonists in schizophrenia - osanetant and talnetant - have shown significant improvement in positive symptoms, with no major side-effects reported as yet. Here we discuss the preclinical and clinical evidence that indicates that NK(3) receptor antagonists might represent a new approach to the treatment of schizophrenia and possibly other neuropsychiatric disorders. PMID- 16341063 TI - Anti-inflammatory effects of statins: clinical evidence and basic mechanisms. AB - Chronic inflammation is a key feature of vascular disease states such as atherosclerosis. Multiple clinical studies have shown that a class of medications termed statins lower cardiovascular morbidity and mortality. Originally developed to lower serum cholesterol, increasing evidence suggests that these medications have potent anti-inflammatory effects that contribute to their beneficial effects in patients. Here, we discuss the clinical and experimental evidence underlying the anti-inflammatory effects of these agents. PMID- 16341064 TI - Exploiting the PI3K/AKT pathway for cancer drug discovery. AB - Evolving studies with several different targeted therapeutic agents are demonstrating that patients with genomic alterations of the target, including amplification, translocation and mutation, are more likely to respond to the therapy. Recent studies indicate that numerous components of the phosphatidylinositol-3-kinase (PI3K)/AKT pathway are targeted by amplification, mutation and translocation more frequently than any other pathway in cancer patients, with resultant activation of the pathway. This warrants exploiting the PI3K/AKT pathway for cancer drug discovery. PMID- 16341065 TI - Finding new tricks for old drugs: an efficient route for public-sector drug discovery. AB - With the annotation of the human genome approaching completion, public-sector researchers - spurred in part by various National Institutes of Health Roadmap Initiatives - have become increasingly engaged in drug discovery and development efforts. Although large and diverse chemical libraries of 'drug-like' compounds can be readily screened to yield chemically novel scaffolds, transforming these 'chemical probes' into drugs is a daunting endeavour. A more efficient approach involves screening libraries of approved and off-patent medications; both phenotypic- and molecular target-based screening of 'old drugs' can readily yield compounds that could be immediately used in clinical trials. Using case studies, we describe how this approach has rapidly identified candidate medications suitable for clinical trials in disorders such as progressive multifocal leukoencephalopathy and amyotrophic lateral sclerosis. This approach has also led to the discovery of the molecular targets responsible for serious drug side effects, thereby allowing efficient 'counter-screening' to avoid these side effects. PMID- 16341066 TI - NAAG peptidase inhibitors and their potential for diagnosis and therapy. AB - Modulation of N-acetyl-L-aspartyl-L-glutamate peptidase activity with small molecule inhibitors holds promise for a wide variety of diseases that involve glutamatergic transmission, and has implications for the diagnosis and therapy of cancer. This new class of compounds, of which at least one has entered clinical trials and proven to be well tolerated, has demonstrated efficacy in experimental models of pain, schizophrenia, amyotrophic lateral sclerosis, traumatic brain injury and, when appropriately functionalized, can image prostate cancer. Further investigation of these promising drug candidates will be needed to bring them to the marketplace. The recent publication of the X-ray crystal structure for the enzymatic target of these compounds should facilitate the development of other new agents with enhanced activity that could improve both the diagnosis and treatment of neurological disorders. PMID- 16341069 TI - The genesis and evolution of homeobox gene clusters. AB - Once called the 'Rosetta stone' of developmental biology, the homeobox continues to fascinate both evolutionary and developmental biologists. The birth of the homeotic, or Hox, gene cluster, and its subsequent evolution, has been crucial in mediating the major transitions in metazoan body plan. Comparative genomics studies indicate that the more recently discovered ParaHox and NK clusters were linked to the Hox cluster early in evolution, and that together they constituted a 'megacluster' of homeobox genes that conspicuously contributed to body-plan evolution. PMID- 16341070 TI - Modulating Hox gene functions during animal body patterning. AB - With their power to shape animal morphology, few genes have captured the imagination of biologists as the evolutionarily conserved members of the Hox clusters have done. Recent research has provided new insight into how Hox proteins cause morphological diversity at the organismal and evolutionary levels. Furthermore, an expanding collection of sequences that are directly regulated by Hox proteins provides information on the specificity of target-gene activation, which might allow the successful prediction of novel Hox-response genes. Finally, the recent discovery of microRNA genes within the Hox gene clusters indicates yet another level of control by Hox genes in development and evolution. PMID- 16341071 TI - Arthropod segmentation: beyond the Drosophila paradigm. AB - Most of our knowledge about the mechanisms of segmentation in arthropods comes from work on Drosophila melanogaster. In recent years it has become clear that this mechanism is far from universal, and different arthropod groups have distinct modes of segmentation that operate through divergent genetic mechanisms. We review recent data from a range of arthropods, identifying which features of the D. melanogaster segmentation cascade are present in the different groups, and discuss the evolutionary implications of their conserved and divergent aspects. A model is emerging, although slowly, for the way that arthropod segmentation mechanisms have evolved. PMID- 16341072 TI - The evolution of metazoan axial properties. AB - Renewed interest in the developmental basis of organismal complexity, and the emergence of new molecular tools, is improving our ability to study the evolution of metazoan body plans. The most substantial changes in body-plan organization occurred early in metazoan evolution; new model systems for studying basal metazoans are now being developed, and total-genome-sequencing initiatives are underway for at least three of the four most important taxa. The elucidation of how the gene networks that are involved in axial organization, germ-layer formation and cell differentiation are used differently during development is generating a more detailed understanding of the events that have led to the current diversity of multicellular life. PMID- 16341073 TI - Cilia and centrosomes: a unifying pathogenic concept for cystic kidney disease? AB - Cystic kidney diseases are among the most frequent lethal genetic diseases. Positional cloning of novel cystic kidney disease genes revealed that their products (cystoproteins) are expressed in sensory organelles called primary cilia, in basal bodies or in centrosomes. Primary cilia link mechanosensory, visual, osmotic, gustatory and other stimuli to mechanisms of cell-cycle control and epithelial cell polarity. The ciliary expression of cystoproteins explains why many other organs might be also affected in patients with cystic kidney disease. Protein-protein interactions among cystoproteins, and their strong evolutionary conservation, provide a basis for a multidisciplinary approach to unravelling the novel signalling mechanisms that are involved in this disease group. PMID- 16341074 TI - Timeline: Hermann Joseph Muller, evolutionist. AB - This essay is dedicated to the proposition that Hermann Joseph Muller, widely regarded as the greatest geneticist of the first half-century of the subject, was also one of the greatest evolutionists of this period. His Nobel Prize-winning work, which showed that radiation increases the mutation rate, is in every genetics textbook, and his prescient ideas have influenced almost every aspect of the discipline. Here I emphasize his less well-known contribution to the neo Darwinian theory of evolution. PMID- 16341075 TI - Science and society: protecting crop genetic diversity for food security: political, ethical and technical challenges. AB - Crop genetic diversity - which is crucial for feeding humanity, for the environment and for sustainable development - is being lost at an alarming rate. Given the enormous interdependence of countries and generations on this genetic diversity, this loss raises critical socio-economic, ethical and political questions. The recent ratification of a binding international treaty, and the development of powerful new technologies to conserve and use resources more effectively, have raised expectations that must now be fulfilled. PMID- 16341078 TI - Cleavage pattern and emerging asymmetry of the mouse embryo. AB - Early mammalian development is regulative - it is flexible and responsive to experimental intervention. This flexibility could be explained if embryogenesis were originally completely unbiased and disordered; order and determination of cells only arising later. Alternatively, regulative behaviour could be consistent with the embryo having some order or bias from the very beginning, with inflexibility and cell determination increasing steadily over time. Recent evidence supports the second view and indicates that the sequence and the orientations of cell divisions help to build the first asymmetries. PMID- 16341079 TI - Kinetochore capture and bi-orientation on the mitotic spindle. AB - Kinetochores are large protein complexes that are formed on chromosome regions known as centromeres. For high-fidelity chromosome segregation, kinetochores must be correctly captured on the mitotic spindle before anaphase onset. During prometaphase, kinetochores are initially captured by a single microtubule that extends from a spindle pole and are then transported poleward along the microtubule. Subsequently, microtubules that extend from the other spindle pole also interact with kinetochores and, eventually, each sister kinetochore attaches to microtubules that extend from opposite poles - this is known as bi orientation. Here we discuss the molecular mechanisms of these processes, by focusing on budding yeast and drawing comparisons with other organisms. PMID- 16341080 TI - Suffering in silence: the tolerance of DNA damage. AB - When cells that are actively replicating DNA encounter sites of base damage or strand breaks, replication might stall or arrest. In this situation, cells rely on DNA-damage-tolerance mechanisms to bypass the damage effectively. One of these mechanisms, known as translesion DNA synthesis, is supported by specialized DNA polymerases that are able to catalyse nucleotide incorporation opposite lesions that cannot be negotiated by high-fidelity replicative polymerases. A second category of tolerance mechanism involves alternative replication strategies that obviate the need to replicate directly across sites of template-strand damage. PMID- 16341081 TI - Arp2/3 and SCAR: plants move to the fore. AB - The actin-nucleating Arp2/3 complex is essential for life in yeast and animals, but not in plants, in which mutants of Arp2/3 complex components show relatively minor developmental abnormalities. Animal cells control the activity of the Arp2/3 complex through the suppressor of cyclic AMP receptor (SCAR) complex to achieve cell motility. Amazingly, plants have also retained the SCAR cell motility pathway, and now provide a unique model for the study of new aspects of SCAR function in the absence of cell motility. PMID- 16341082 TI - Circadian clocks - the fall and rise of physiology. AB - Circadian clocks control the daily life of most light-sensitive organisms - from cyanobacteria to humans. Molecular processes generate cellular rhythmicity, and cellular clocks in animals coordinate rhythms through interaction (known as coupling). This hierarchy of clocks generates a complex, approximately 24-hour temporal programme that is synchronized with the rotation of the Earth. The circadian system ensures anticipation and adaptation to daily environmental changes, and functions on different levels - from gene expression to behaviour. Circadian research is a remarkable example of interdisciplinarity, unravelling the complex mechanisms that underlie a ubiquitous biological programme. Insights from this research will help to optimize medical diagnostics and therapy, as well as adjust social and biological timing on the individual level. PMID- 16341083 TI - Oncogenic PI3K deregulates transcription and translation. AB - There have long been indications of a role for PI3K (phosphatidylinositol 3 kinase) in cancer pathogenesis. Experimental data document a requirement for deregulation of both transcription and translation in PI3K-mediated oncogenic transformation. The recent discoveries of cancer-specific mutations in PIK3CA, the gene that encodes the catalytic subunit p110alpha of PI3K, have heightened the interest in the oncogenic potential of this lipid kinase and have made p110alpha an ideal drug target. PMID- 16341084 TI - Statins and cancer prevention. AB - Randomized controlled trials for preventing cardiovascular disease indicated that statins had provocative and unexpected benefits for reducing colorectal cancer and melanoma. These findings have led to the intensive study of statins in cancer prevention, including recent, large population-based studies showing statin associated reductions in overall, colorectal and prostate cancer. Understanding the complex cellular effects (for example, on angiogenesis and inflammation) and the underlying molecular mechanisms of statins (for example, 3-hydroxy-3 methylglutaryl coenzyme-A (HMG-CoA) reductase-dependent processes that involve geranylgeranylation of Rho proteins, and HMG-CoA-independent processes that involve lymphocyte-function-associated antigen 1) will advance the development of molecularly targeted agents for preventing cancer. This understanding might also help the development of drugs for other ageing-related diseases with interrelated molecular pathways. PMID- 16341085 TI - A candidate gene approach to searching for low-penetrance breast and prostate cancer genes. AB - Most cases of breast and prostate cancer are not associated with mutations in known high-penetrance genes, indicating the involvement of multiple low penetrance risk alleles. Studies that have attempted to identify these genes have met with limited success. The National Cancer Institute Breast and Prostate Cancer Cohort Consortium--a pooled analysis of multiple large cohort studies with a total of more than 5,000 cases of breast cancer and 8,000 cases of prostate cancer--was therefore initiated. The goal of this consortium is to characterize variations in approximately 50 genes that mediate two pathways that are associated with these cancers--the steroid-hormone metabolism pathway and the insulin-like growth factor signalling pathway--and to associate these variations with cancer risk. PMID- 16341086 TI - Science and society: the Long Island Breast Cancer Study Project. AB - In the early 1990s, breast cancer advocates petitioned the United States Congress to investigate the high rates of breast cancer on Long Island in the state of New York. The resulting law led to the Long Island Breast Cancer Study Project (LIBCSP)--more than ten research projects designed to study the possible causes of this increased incidence of cancer. This project reported that there was no evidence that environmental exposures were responsible. Controversial from its start, the LIBCSP has had an important role in efforts to understand the reasons for the high rates of breast cancer in some regions of the United States. PMID- 16341087 TI - Ion-binding properties of the ClC chloride selectivity filter. AB - The ClC channels are members of a large protein family of chloride (Cl-) channels and secondary active Cl- transporters. Despite their diverse functions, the transmembrane architecture within the family is conserved. Here we present a crystallographic study on the ion-binding properties of the ClC selectivity filter in the close homolog from Escherichia coli (EcClC). The ClC selectivity filter contains three ion-binding sites that bridge the extra- and intracellular solutions. The sites bind Cl- ions with mM affinity. Despite their close proximity within the filter, the three sites can be occupied simultaneously. The ion-binding properties are found conserved from the bacterial transporter EcClC to the human Cl- channel ClC-1, suggesting a close functional link between ion permeation in the channels and active transport in the transporters. In resemblance to K+ channels, ions permeate the ClC channel in a single file, with mutual repulsion between the ions fostering rapid conduction. PMID- 16341088 TI - Intracellular calcium changes trigger connexin 32 hemichannel opening. AB - Connexin hemichannels have been proposed as a diffusion pathway for the release of extracellular messengers like ATP and others, based on connexin expression models and inhibition by gap junction blockers. Hemichannels are opened by various experimental stimuli, but the physiological intracellular triggers are currently not known. We investigated the hypothesis that an increase of cytoplasmic calcium concentration ([Ca2+]i) triggers hemichannel opening, making use of peptides that are identical to a short amino-acid sequence on the connexin subunit to specifically block hemichannels, but not gap junction channels. Our work performed on connexin 32 (Cx32)-expressing cells showed that an increase in [Ca2+]i triggers ATP release and dye uptake that is dependent on Cx32 expression, blocked by Cx32 (but not Cx43) mimetic peptides and a calmodulin antagonist, and critically dependent on [Ca2+]i elevation within a window situated around 500 nM. Our results indicate that [Ca2+]i elevation triggers hemichannel opening, and suggest that these channels are under physiological control. PMID- 16341089 TI - Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. AB - Mitochondria are indispensable for cell viability; however, major mitochondrial functions including citric acid cycle and oxidative phosphorylation are dispensable. Most known essential mitochondrial proteins are involved in preprotein import and assembly, while the only known essential biosynthetic process performed by mitochondria is the biogenesis of iron-sulfur clusters (ISC). The components of the mitochondrial ISC-assembly machinery are derived from the prokaryotic ISC-assembly machinery. We have identified an essential mitochondrial matrix protein, Isd11 (YER048w-a), that is found in eukaryotes only. Isd11 is required for biogenesis of cellular Fe/S proteins and thus is a novel subunit of the mitochondrial ISC-assembly machinery. It forms a complex with the cysteine desulfurase Nfs1 and is required for formation of an Fe/S cluster on the Isu scaffold proteins. We conclude that Isd11 is an indispensable eukaryotic component of the mitochondrial machinery for biogenesis of Fe/S proteins. PMID- 16341090 TI - The Nfs1 interacting protein Isd11 has an essential role in Fe/S cluster biogenesis in mitochondria. AB - Formation of iron/sulfur (Fe/S) clusters, protein translocation and protein folding are essential processes in the mitochondria of Saccharomyces cerevisiae. In a systematic approach to characterize essential proteins involved in these processes, we identified a novel essential protein of the mitochondrial matrix, which is highly conserved from yeast to human and which we termed Isd11. Depletion of Isd11 caused a strong reduction in the levels of the Fe/S proteins aconitase and the Rieske protein, and a massive decrease in the enzymatic activities of aconitase and succinate dehydrogenase. Incorporation of iron into the Fe/S protein Leu1 and formation of the Fe/S cluster containing holoform of the mitochondrial ferredoxin Yah1 were inhibited in the absence of Isd11. This strongly suggests that Isd11 is required for the assembly of Fe/S proteins. We show that Isd11 forms a stable complex with Nfs1, the cysteine desulfurase of the mitochondrial machinery for Fe/S cluster assembly. In the absence of Isd11, Nfs1 is prone to aggregation. We propose that Isd11 acts together with Nfs1 in an early step in the biogenesis of Fe/S proteins. PMID- 16341091 TI - Integron cassette insertion: a recombination process involving a folded single strand substrate. AB - Integrons play a major role in the dissemination of antibiotic resistance genes among Gram-negative pathogens. Integron gene cassettes form circular intermediates carrying a recombination site, attC, and insert into an integron platform at a second site, attI, in a reaction catalyzed by an integron-specific integrase IntI. The IntI1 integron integrase preferentially binds to the 'bottom strand' of single-stranded attC. We have addressed the insertion mechanism in vivo using a recombination assay exploiting plasmid conjugation to exclusively deliver either the top or bottom strand of different integrase recombination substrates. Recombination of a single-stranded attC site with an attI site was 1000-fold higher for one strand than for the other. Conversely, following conjugative transfer of either attI strand, recombination with attC is highly unfavorable. These results and those obtained using mutations within a putative attC stem-and-loop strongly support a novel integron cassette insertion model in which the single bottom attC strand adopts a folded structure generating a double strand recombination site. Thus, recombination would insert a single strand cassette, which must be subsequently processed. PMID- 16341092 TI - Different HECT domain ubiquitin ligases employ distinct mechanisms of polyubiquitin chain synthesis. AB - Individual ubiquitin (Ub)-protein ligases (E3s) cooperate with specific Ub conjugating enzymes (E2s) to modify cognate substrates with polyubiquitin chains. E3s belonging to the Really Interesting New Gene (RING) and Homologous to E6 Associated Protein (E6AP) C-Terminus (HECT) domain families utilize distinct molecular mechanisms. In particular, HECT E3s, but not RING E3s, form a thiol ester with Ub before transferring Ub to the substrate lysine. Here we report that different HECT domain E3s can employ distinct mechanisms of polyubiquitin chain synthesis. We show that E6AP builds up a K48-linked chain on its HECT cysteine residue, while KIAA10 builds up K48- and K29-linked chains as free entities. A small region near the N-terminus of the conserved HECT domain helps to bring about this functional distinction. Thus, a given HECT domain can specify both the linkage of a polyubiquitin chain and the mechanism of its assembly. PMID- 16341094 TI - Herceptin: HERalding a new era in breast cancer care but at what cost? PMID- 16341093 TI - Activation or suppression of NFkappaB by HPK1 determines sensitivity to activation-induced cell death. AB - Restimulation of the T-cell receptor (TCR) in activated T cells induces CD95 (Fas/Apo-1)-mediated activation-induced cell death (AICD). The TCR-proximal mechanisms leading to AICD are elusive. Here we characterize hematopoietic progenitor kinase 1 (HPK1) as a differentially regulated TCR-proximal signaling protein involved in AICD of primary T cells. We show that HPK1 is a functional component of the endogenous IkappaB kinase (IKK) complex and is crucial for TCR mediated NFkappaB activation. While full-length HPK1 enhances IKKbeta phosphorylation, siRNA-mediated knockdown of HPK1 blunts TCR-mediated NFkappaB activation and increases cell death. We also demonstrate proteolytic processing of HPK1 into HPK1-C, specifically in AICD-sensitive primary T cells. The cleavage product HPK1-C sequesters the inactive IKK complex and suppresses NFkappaB upon TCR restimulation by binding to IKKalpha and IKKbeta. T cells of HPK1-C transgenic mice are sensitized towards TCR-mediated AICD. Consequently, preventing HPK1-C generation in primary T cells by siRNA-mediated knockdown results in decreased AICD. Thus, these results show a novel mechanism of sensitization of T lymphocytes towards AICD by suppression of NFkappaB, and propose that HPK1 is a life/death switch in T lymphocytes. PMID- 16341095 TI - The evolving role of aromatase inhibitors in the treatment of early-stage breast cancer. PMID- 16341096 TI - Individualization of neoadjuvant therapy for breast cancer according to molecular tumor characteristics. PMID- 16341111 TI - Optimizing the sequencing of adjuvant chemotherapy and radiotherapy for early breast cancer. PMID- 16341112 TI - Is radical prostatectomy of benefit in men with localized prostate cancer? PMID- 16341113 TI - Can dose-dense chemotherapy improve outcome in patients with better-prognosis small-cell lung cancer? PMID- 16341114 TI - Do low-molecular-weight heparins improve the survival of cancer patients? PMID- 16341115 TI - Does radiotherapy to the head, neck or upper thorax for childhood cancer increase the risk of later thyroid cancer? PMID- 16341116 TI - Does lung cancer screening motivate smoking cessation? PMID- 16341117 TI - Evolving paradigms and perceptions of cancer. AB - The word cancer produces widely differing perceptions between the general public, and the scientific and medical communities. These different ideas lead to very diverse understandings of the disease. The paradigms affect both the focus and design of research and also impact upon patient care. The cultural perception is very pessimistic: a relentless, incurable, extremely painful disease, the treatment of which is conceived as difficult, with little chance of a simple cure. Within the medical and scientific communities, however, there are a number of quite different views of the disease. Both the orderly extension of disease described by Halsted, and the systemic nature of cancer even when it appears to be localized, are perceptions within the professional community. The promise of a 'magic bullet' is in sharp contrast to the incremental advances seen in clinical oncology. What is needed is a clear recognition of how these varying perceptions of cancer affect and limit communication among the cancer-related disciplines as well as between these disciplines and the public. Both professionals and the general public should consider cancer as a group of diseases for which cure is related to tumor type, stage and available treatment. PMID- 16341118 TI - Epidemiology informing clinical practice: from bills of mortality to population laboratories. AB - The earliest observations on population patterns of disease and how they might inform medical practice probably occurred during the 17th century, and they continue to the present day, with increasing relevance to nutritional and infectious diseases, and cancer and other chronic diseases. Chronic-disease methods grew out of infectious-disease epidemiology, in which both field and laboratory methods are used. In diseases where intermediate biology was not initially observable (particularly cancer), record-based and interview-based epidemiology revealed some key exposures (e.g. smoking and radiation). With measurable intermediates (e.g. blood lipids), cardiovascular epidemiology also yielded inferences on causal pathways. Important changes that are remaking the field of epidemiology and will ultimately influence all aspects of medical practice include the following: high-throughput genotyping, allowing genetic and gene-environment causes of disease to be identified; high-throughput proteomics, which should allow the development of early-detection methods; new tools for the measurement of exposures; and a molecular basis for disease taxonomy. These new methods will allow a much better understanding of both the etiology and the intermediate stages of disease; however, new methods do not obviate the necessity for good study design, especially the need to be clear on the difference between observation and experiment. The greatest opportunities to inform medical practice come from the application of new methods to large-scale human observational studies, which include genetics, environment, early-detection markers, molecular classification of outcome, and treatment data. Improved molecular classification of disease will allow smaller, focused clinical trials to be undertaken and, ultimately, the tailoring of treatment to the biological profile of patient and disease. PMID- 16341119 TI - Mechanisms of Disease: prediction and prevention of breast cancer--cellular and molecular interactions. AB - Breast cancer is the most prevalent female cancer in the world and its incidence is increasing, largely because of the Western lifestyle. There is a need, not only to predict women who will develop the disease, but also to apply drug and lifestyle measures in order to prevent the disease. Current risk prediction models are based on combinations of risk factors and have good predictive but low discriminatory power. New risk prediction methods might come from examination of single nucleotide polymorphisms in several genes or from an increased knowledge of the molecular and cellular biology of the breast, particularly with respect to aberrant gene expression and protein synthesis. These methods might also determine new targets for preventive agents and lifestyle change. Many potential preventive measures are available and some have been successful. New approaches are required, however, not only to prevent the disease but to devise methods for their assessment that do not require very large and expensive clinical trials. PMID- 16341120 TI - Successful pregnancies following fertility-preserving treatment for ovarian carcinoma. AB - BACKGROUND: A 30-year-old woman underwent a routine sonogram at 6 weeks of gestation during her first pregnancy and was found to have a 6 cm complex right ovarian mass, thought to be a dermoid cyst. The pregnancy progressed to term and the patient went on to have a normal vaginal delivery. At 3 months postpartum, she underwent a laparoscopic ovarian cystectomy. Preoperative serum cancer antigen 125 was within the normal range at 23.6 U/ml. INVESTIGATIONS: Pelvic ultrasound, evaluation of serum cancer antigen 125, abdominal and pelvic CT scans. DIAGNOSIS: Stage IA clear-cell epithelial carcinoma of the right ovary. MANAGEMENT: Laparoscopic ovarian cystectomy, dilation and curettage, laparotomy, right salpingo-oophorectomy, infracolic omentectomy, peritoneal biopsy, peritoneal washing, appendectomy, bilateral pelvic and para-aortic lymphadenectomy, adjuvant chemotherapy with paclitaxel and carboplatin. PMID- 16341122 TI - PS externalization: from corpse clearance to drug delivery. PMID- 16341121 TI - High Mobility Group A1 (HMGA1) proteins interact with p53 and inhibit its apoptotic activity. AB - HMGA gene overexpression and rearrangements are frequent in several tumours, but their oncogenic function is still unclear. Here we report of a physical and functional interaction between High Mobility Group A1 (HMGA1) protein and p53 oncosuppressor. We found that HMGA1 binds p53 in vitro and in vivo, and both proteins are present in the same complexes bound to the Bax gene promoter. HMGA1 interferes with the p53-mediated transcription of p53 effectors Bax and p21(waf1) while cooperates with p53 in the transcriptional activation of the p53 inhibitor mdm2. This transcriptional modulation is associated with a reduced p53-dependent apoptosis in cells expressing exogenous HMGA1 and p53, or in cells expressing endogenously the proteins and in which p53 was activated by UV-irradiation. Furthermore, antisense inhibition of HMGA1b expression dramatically increases the UV-induced p53-mediated apoptosis. These data define a new physical and functional interaction between HMGA1 and p53 that modulates transcription of p53 target genes and inhibits apoptosis. PMID- 16341123 TI - Recognition of apoptotic cells by macrophages activates the peroxisome proliferator-activated receptor-gamma and attenuates the oxidative burst. AB - It is appreciated that phagocytosis of apoptotic cells (AC) is an immunological relevant process that shapes the pro- versus anti-inflammatory macrophage phenotype. It was our intention to study the respiratory burst, a prototype marker of macrophage activation, under the impact of AC. Following incubation of RAW264.7 macrophages with AC, we noticed attenuated production of reactive oxygen species (ROS) in response to PMA treatment, and observed a correlation between attenuated ROS formation and suppression of protein kinase Calpha (PKCalpha) activation. EMSA analysis demonstrated an immediate activation of peroxisome proliferator-activated receptor-gamma (PPARgamma) following supplementation of AC to macrophages. In macrophages carrying a dominant-negative PPARgamma mutant, recognition of AC no longer suppressed PKCalpha activation, and the initial phase of ROS formation was largely restored. Interference with actin polymerization and transwell experiments suggest that recognition of AC by macrophages suffices to attenuate the early phase of ROS formation that is attributed to PPARgamma activation. PMID- 16341124 TI - Reactive oxygen species mediate crosstalk between NF-kappaB and JNK. AB - The activation of NF-kappaB inhibits apoptosis via a mechanism involving upregulation of various antiapoptotic genes, such as cellular FLICE-inhibitory protein (c-FLIP), Bcl-xL, A1/Bfl-1, and X chromosome-liked inhibitor of apoptosis (XIAP). In contrast, the activation of c-Jun N-terminal kinase (JNK) promotes apoptosis in a manner that is dependent on the cell type and the context of the stimulus. Recent studies have indicated that one of the antiapoptotic functions of NF-kappaB is to downregulate JNK activation. Further studies have also revealed that NF-kappaB inhibits JNK activation by suppressing accumulation of reactive oxygen species (ROS). In this review, we will focus on the signaling crosstalk between the NF-kappaB and JNK cascades via ROS. PMID- 16341125 TI - Small molecule inhibitors of Apaf-1-related caspase- 3/-9 activation that control mitochondrial-dependent apoptosis. AB - Apoptosis is a biological process relevant to human disease states that is strongly regulated through protein-protein complex formation. These complexes represent interesting points of chemical intervention for the development of molecules that could modulate cellular apoptosis. The apoptosome is a holoenzyme multiprotein complex formed by cytochrome c-activated Apaf-1 (apoptotic protease activating factor), dATP and procaspase-9 that link mitochondria disfunction with activation of the effector caspases and in turn is of interest for the development of apoptotic modulators. In the present study we describe the identification of compounds that inhibit the apoptosome-mediated activation of procaspase-9 from the screening of a diversity-oriented chemical library. The active compounds rescued from the library were chemically optimised to obtain molecules that bind to both recombinant and human endogenous Apaf-1 in a cytochrome c-noncompetitive mechanism that inhibits the recruitment of procaspase 9 by the apoptosome. These newly identified Apaf-1 ligands decrease the apoptotic phenotype in mitochondrial-mediated models of cellular apoptosis. PMID- 16341126 TI - Notch1 confers thymocytes a resistance to GC-induced apoptosis through Deltex1 by blocking the recruitment of p300 to the SRG3 promoter. AB - One notable phenotypic change during the differentiation of immature thymocytes into either mature CD4 or CD8 single-positive lineages is the acquisition of a resistance to glucocorticoid (GC)-induced apoptosis. We have previously reported that SRG3 is critical in determining the sensitivity for the GC-induced apoptosis in developing thymocytes. We report here that Notch signaling downregulates the transcriptional activation of SRG3 through N-box and/or E-box elements on its promoter. RBP-J represses SRG3 transcription through the N-box motif. On the other hand, Deltex1 competitively inhibits the binding of p300 to E2A/HEB protein bound to the E-box elements and represses the SRG3 promoter activity. Moreover, enforced expression of Deltex1 restored double-positive (DP) thymocyte survival from the GC-induced apoptosis. Our results suggest that Notch signaling confers differentiating DP thymocytes resistance to GCs by regulating the SRG3 expression through Deltex1, and that Deltex1 and SRG3 may play a significant role during DP thymocyte maturation. PMID- 16341127 TI - Tumor suppressor pp32 represses cell growth through inhibition of transcription by blocking acetylation and phosphorylation of histone H3 and initiating its proapoptotic activity. AB - pp32 belongs to a family of leucine-rich acidic nuclear proteins, which play important roles in many cellular processes including regulation of chromatin remodeling, transcription, RNA transport, transformation and apoptosis. pp32 is described as a new tumor suppressor. It is unknown as to how pp32 works in tumor suppression. We found that overexpression of pp32 in human Jurkat T cells inhibits cell growth, and silenced pp32 promotes growth. We first showed that hyperacetylation and hyperphosphorylation of histone H3 are required for T-cell activation. Phosphorylation of histone H3 precedes acetylation during T-cell activation. pp32 specifically binds to histone H3 and blocks its acetylation and phosphorylation. pp32 directly initiates caspase activity and also promotes granzyme A-mediated caspase-independent cell death. Taken together, pp32 plays a repressive role by inhibiting transcription and triggering apoptosis. PMID- 16341128 TI - Overexpression of RARgamma increases death of SH-SY5Y neuroblastoma cells in response to retinoic acid but not fenretinide. PMID- 16341129 TI - Functional observations in vitamin A deficiency: diagnosis and time course of recovery. AB - AIMS: To describe the effects of vitamin A deficiency (VAD) on retinal function and the subsequent recovery following treatment in three patients with systemic conditions (two with Crohn disease; one secondary to IgE syndrome). METHODS: Electrophysiological testing (including pattern electroretinogram, PERG; electroretinogram, ERG; visual-evoked potential) established the diagnosis of VAD. Repeat testing was carried out in two patients to monitor the time course of recovery following intramuscular vitamin A injection. The third patient had repeat recordings following 13 months of oral supplementation. RESULTS: All three patients initially displayed a characteristic absence of rod function associated with VAD. In addition, delayed and reduced amplitude cone ERGs, loss of short wavelength cone (S-cone) function and subnormal macular function were observed in two patients. Restoration of rod and generalised cone function was rapid in the two patients who received intramuscular injection, with normalisation of some electrophysiological responses after only 3 days. Normal S-cone amplitudes and cone latencies were reached within 12 days of vitamin A injection. Macular function returned to within normal limits by 12 days postinjection in one patient, but remained mildly subnormal in the second patient. Full recovery was present after 13 months oral supplementation in the third patient. CONCLUSIONS: Novel observations regarding dark-adapted cone function, S-cone function, and PERG are presented. The differences between the effects of VAD on rod and cone function, and their rate of recovery, may reflect differences in the visual cycle between the two photoreceptor classes. The importance of rapidly and accurately diagnosing VAD, a treatable condition, is noted. PMID- 16341131 TI - The surgical management of chronic hypotony due to uveitis. PMID- 16341132 TI - Bilateral dacroadenitis associated with acute Crohn's disease. PMID- 16341133 TI - Tractional retinal break and rhegmatogenous retinal detachment consequent to branch retinal vein occlusion. PMID- 16341134 TI - Serratia marcescens endophthalmitis secondary to pneumonia. PMID- 16341135 TI - Accuracy of GDx variable corneal compensation polarization measurements in normal human eyes: effect of accommodation, cycloplegia, focus, pupil size, and eye selection on reproducibility. AB - PURPOSE: To evaluate the reproducibility of variable corneal compensation (VCC) and the effect of accommodation, cycloplegia, eye selection, focus, and pupil size on this measurement of polarization. METHODS: Using a GDx scanning laser polarimeter, multiple measurements of the VCC were obtained from each eye of 33 healthy, young adults under differing conditions. Pupil size and refraction were independently measured with a pupillometer and an autorefractometer. The effects of eye, instrument focus, pupil diameter, cycloplegia, and accommodation were statistically assessed. RESULTS: The reproducibility of a single retardation measurement and its axis, as determined by the standard deviations (SD) of repeated measurements, is +/-1.58 nm and 2.10 degrees , respectively. There is a difference in retardation between right and left eyes, of 5.26+/-9 nm, P=0.002. Increasing pupil size increases retardation. Cycloplegia or defocusing decreases retardation, and pharmacologically induced accommodation has no effect on retardation. CONCLUSIONS: The retardation and its axis are highly reproducible measurements when the pupil is of physiologic size and the GDx is properly focused. There is a consistent difference in VCC retardation between the paired right and left eyes. This difference may reflect equipment-induced measurement artefact and/or an anatomic asymmetry between the paired eyes of the subjects studied. Clinicians should be cautious when comparing interocular VCC measurements between paired right and left eyes and using data pooled from both eyes for age-adjusted, normalized standards. PMID- 16341136 TI - Tear function and goblet cell density after pterygium excision. AB - PURPOSE: To evaluate the short-term effect of pterygium excision on tear function and conjunctival goblet cell density. METHODS: In all, 70 patients (70 eyes) with primary pterygia who underwent a bare-sclera procedure were enrolled in this study. Tear break-up time, Schirmer, and tear-ferning tests and conjunctival goblet cell density obtained by impression cytology were evaluated before and 1 month after surgery. RESULTS: At 1 month postoperation, the tear break-up time (11.49+/-3.76 s) was significantly prolonged when compared with the preoperation tear break-up time (9.74+/-3.43 s, P=0.002). Before surgery, only 17% of the patients showed normal crystallization in the tear-ferning test. This proportion was significantly increased to 90% 1 month after surgery (P<0.001). There is no significant difference in the Schirmer test value between pre- and postsurgery (P>0.05). The mean goblet cell density in conjunctival impression specimens before surgery was 41.82+/-18.29/10 fields, which was increased significantly 1 month after surgery (P<0.001) with a mean cell density of 50.67+/-18.71/10 fields. CONCLUSION: Tear function in patients with primary pterygium improves after pterygium excision, which indicates that pterygium has a close relationship with dry eye. PMID- 16341137 TI - An unusual case of proptosis. PMID- 16341138 TI - Methylprednisolone pulse therapy in patient with isolated superior oblique myositis. PMID- 16341139 TI - Dectin-1: a signalling non-TLR pattern-recognition receptor. AB - Dectin-1 is a natural killer (NK)-cell-receptor-like C-type lectin that is thought to be involved in innate immune responses to fungal pathogens. This transmembrane signalling receptor mediates various cellular functions, from fungal binding, uptake and killing, to inducing the production of cytokines and chemokines. These activities could influence the resultant immune response and can, in certain circumstances, lead to autoimmunity and disease. As I discuss here, understanding the molecular mechanisms behind these functions has revealed new concepts, including collaborative signalling with the Toll-like receptors (TLRs) and the use of spleen tyrosine kinase (SYK), that have implications for the role of other non-TLR pattern-recognition receptors in immunity. PMID- 16341140 TI - Papillomaviruses as targets for cancer gene therapy. AB - The human papillomaviruses (HPVs) are a diverse group of infectious agents, some of which are a causative agent of human cancers. Cervical cancer and oral cancer are closely associated with specific types of HPV, and the tumors grow only if there is continual expression of the viral E6 and E7 genes. Evidence from in vitro studies shows that when expression of these genes is inhibited by gene therapy approaches such as antisense RNA, ribozymes, or siRNA, the transformed phenotype of the cells is lost. Although it seems possible that clinical applications of this approach could help in the management of cervical and oral cancers there have been no clinical trials of gene therapy for HPV-associated cancers. Since the basic information is now available, a shift to translational research would be greatly welcomed. PMID- 16341141 TI - Selective replication of E1B55K-deleted adenoviruses depends on enhanced E1A expression in cancer cells. AB - E1B55K-deleted dl1520 could selectively replicate in cancer cells and has been used in clinical trials as an antitumor agent. The mechanism of virus selective replication in cancer cells, including a possible role of p53, is unclear. Studies with established cancer cell lines have demonstrated that some cancer cells are resistant to dl1520 replication, regardless of the p53 status. Hep3B cells supported the E1b-deleted adenoviruses to replicate, whereas Saos2 cells were resistant to viral replication. We applied p53-null Hep3B and Saos2 cells as models to clarify the replication ability of E1B55K-deleted adenoviruses with different expression levels of E1a. We show that lower E1A expression in Saos2 may be the reason for the poor replication in some cancer cells due to the fact that E1a promoter was less activated in Saos2 than in Hep3B. We also demonstrate that the E1B55K protein can increase E1A expression in Saos2 cells for efficient virus replication. In addition, the upstream regions of the E1a promoter have transcriptional activity in Hep3B cells but not in Saos2 cells. The viral E1B55K protein may activate cancer cellular factor(s) that targets the upstream regions of the E1a gene to increase its expression. This is the first study demonstrating that E1B55K protein affects the E1A production levels that is related to cancer selective replication. Our studies have suggested that increase of E1A expression from E1b-deleted adenoviruses may enhance killing cancer cells that otherwise are resistant to viral replication. PMID- 16341142 TI - Peritumoral injection of adenovirus vector expressing NK4 combined with gemcitabine treatment suppresses growth and metastasis of human pancreatic cancer cells implanted orthotopically in nude mice and prolongs survival. AB - NK4 or adenovirus vector expressing NK4 (Ad-NK4) can act bifunctionally as a hepatocyte growth factor antagonist and angiogenesis inhibitor and has potential value in cancer therapy. The aim of this study was to evaluate the therapeutic efficacy of Ad-NK4 in combination with gemcitabine (GEM) against pancreatic cancer. In vitro study showed a strong antiproliferative effect of GEM and a potent anti-invasive effect of Ad-NK4 against pancreatic cancer cells. In in vivo experiments, SUIT-2 human pancreatic cancer cells were implanted into the pancreas of nude mice. Mice were treated with Ad-NK4 by injection into the peritumoral region of the pancreas on day 5 after implantation followed by weekly i.p. injections of GEM. On day 28 after implantation, pancreatic tumor volume was significantly smaller than that in mice treated with Ad-LacZ, Ad-NK4 alone, or GEM alone. Furthermore, combination therapy completely suppressed peritoneal dissemination and liver metastases, leading to significantly increased survival. Histologic and immunohistochemical assays of primary tumors indicated that combination therapy prohibited both cell proliferation and angiogenesis, resulting in high levels of apoptosis. These results suggest that peritumoral injection of Ad-NK4 plus GEM is a potent combination therapy for pancreatic cancer. PMID- 16341143 TI - Th2-dominated antitumor immunity induced by DNA immunization with the genes coding for a basal core peptide PDTRP and GM-CSF. AB - Our previous study showed that DNA vaccination with a plasmid vector encoding a core peptide of mucin1 (PDTRP) provided modest protection against challenge with tumor cells that expressed mucin1 protein. We report here that a DNA vaccine comprising a modified PDTRP plasmid and GM-CSF coding sequence at the C-terminus induced better protection against tumor challenge. The increased protection was directly correlated with a stronger PDTRP-specific immune response induced by the GM-CSF fusion plasmid. The plasmid encoding GM-CSF and the target PDTRP antigen induced a greater PDTRP-specific Th proliferation, antibodies, and cytotoxicity. Interestingly, the modified plasmid vaccine predominantly enhanced the type 2 immune responses manifested by an increased IgG1 to IgG2a antibody ratio and a greater induction of GATA-3 and IL-4 mRNA than that of T-bet and IFN-gamma mRNA in spleen cells from vaccinated mice. In addition, protection against tumor challenge in vaccinated mice showed that there was no significant change in mice survival after in vivo CD8+CTL depletion, indicating that antitumor immunity augmented by plasmid encoding GM-CSF and target PDTRP gene vaccine was dominated by Th2 immune response. PMID- 16341144 TI - Routine immunohistochemical staining for high-molecular weight cytokeratin 34 beta and alpha-methylacyl CoA racemase (P504S) in postirradiation prostate biopsies. AB - A total of 43 cases of postirradiation prostate cores were assessed in an attempt to determine if routine use of alpha-methylacyl-CoA racemase (AMACR) in conjunction with high-molecular weight cytokeratin (HMWCK) would increase the recognition of carcinoma in postirradiation prostate biopsies. We concluded that in most cases the addition of AMACR in conjunction with HMWCK does not increase the recognition of prostatic adenocarcinoma, however it is supportive in nature. In one case the use of AMACR highlighted the extent of the adenocarcinoma which otherwise would have been designated as atypical small acinar proliferation (ASAP). Further evaluation is required to assess the significance of a diagnosis of atypical small acinar proliferation in postirradiation prostate biopsies. PMID- 16341145 TI - Expression of aspartyl beta-hydroxylase and its clinicopathological significance in hepatocellular carcinoma. AB - The human aspartyl beta-hydroxylase is a highly conserved enzyme that hydroxylates epidermal growth factor-like domains in transformation-associated proteins. The aspartyl beta-hydroxylase gene is upregulated in many human malignancies. The purpose of this study was to investigate the expression of aspartyl beta-hydroxylase in hepatocellular carcinoma. Aspartyl beta-hydroxylase mRNA levels were measured in 161 hepatocellular carcinomas and paired nontumorous liver tissues by conventional and real-time RT-PCR. Immunohistochemical staining of aspartyl beta-hydroxylase was performed using EnVision Plus system. The results showed that aspartyl beta-hydroxylase was overexpressed in 150 of 161 hepatocellular carcinomas (93%), including 45 of 48 unifocal small hepatocellular carcinomas (94%). Aspartyl beta-hydroxylase was highly expressed in hepatocellular carcinoma cells in contrast to its low level of expression in non neoplastic liver cells. The protein expression level of aspartyl beta-hydroxylase in the hepatocellular carcinoma was parallel with the mRNA expression level (r=0.6594, P<0.0001). A significantly higher tumor aspartyl beta-hydroxylase overexpression level was associated with the presence of intrahepatic metastasis and the progression of histological grades. In conclusion, aspartyl beta hydroxylase is overexpressed frequently in hepatocellular carcinoma, including early-stage small hepatocellular carcinoma, indicating that overexpression of aspartyl beta-hydroxylase plays a role in the development and progression of hepatocellular carcinoma. PMID- 16341146 TI - Phenotypic evaluation of the basal-like subtype of invasive breast carcinoma. AB - Microarray profiling of invasive breast carcinomas has identified five distinct subtypes of tumors (luminal A, luminal B, normal breast-like, HER2 overexpressing, and basal-like) that are associated with different clinical outcomes. The basal-like subtype is associated with poor clinical outcomes and is the subtype observed in BRCA1-related breast cancers. The aim of this study was to characterize the histologic and immunophenotypic properties of breast basal like carcinomas that were first positively identified using DNA microarray analysis. Detailed histologic review was performed on 56 tumors with known microarray profiles (23 basal-like, 23 luminal, and 12 HER2+). Immunohistochemistry for estrogen receptor (ER), HER2, EGFR, smooth muscle actin (SMA), p63, CD10, cytokeratin 5/6, cytokeratin 8/18, and vimentin was performed on 18 basal-like, 16 luminal, and 12 HER2+ tumors. The basal-like tumors were grade 3 ductal/NOS (21/23) or metaplastic (2/23) carcinomas that frequently showed geographic necrosis (17/23), a pushing border of invasion (14/23), and a stromal lymphocytic response (13/23). Most basal-like tumors showed immunoreactivity for vimentin (17/18), luminal cytokeratin 8/18 (15/18), EGFR (13/18), and cytokeratin 5/6 (11/18), while positivity for the myoepithelial markers SMA (4/18), p63 (4/18) and CD10 (2/18) was infrequent. All basal-like tumors tested were ER- and HER2-. Morphologic features significantly associated with the basal-like subtype included markedly elevated mitotic count (P<0.0001), geographic tumor necrosis (P=0.0003), pushing margin of invasion (P=0.0001), and stromal lymphocytic response (P=0.01). The most consistent immunophenotype seen in the basal-like tumors was negativity for ER and HER2, and positivity for vimentin, EGFR, cytokeratin 8/18, and cytokeratin 5/6. The infrequent expression of myoepithelial markers in basal-like carcinomas does not support a direct myoepithelial cell derivation of these tumors. These findings should further assist in the identification of basal-like carcinomas in clinical specimens, facilitating treatment and epidemiologic studies of this tumor subtype. PMID- 16341147 TI - Cell reservoirs in lymph nodes infected with HIV-1 subtype E differ from subtype B: identification by combined in situ polymerase chain reaction and immunohistochemistry. AB - In Thailand, the predominant HIV subtype is E, rather than subtype B as in North America and Europe. Subtype E has the ability to replicate in vitro in Langerhans cells. We hypothesized that this cell type might constitute a reservoir for the HIV virus in infected lymph nodes. We examined lymph nodes from 25 HIV-1 subtype E-infected patients to determine the immunophenotype of HIV-1-infected cells, their numbers and their distribution. The presence of HIV was detected either by in situ reverse transcriptase-polymerase chain reaction or immunoperoxidase. Cell identity was determined by double labelling using alkaline phosphatase-based immunohistochemistry. The majority of HIV-infected cells in the lymph nodes were Langerhans cells (CD1a+S100+) and Langerhans-related dendritic cells (p55+S100+). These cells were located in the paracortical areas of lymph nodes, with a few cells scattered at the edges of germinal centers, but were absent from germinal centers themselves, in contrast to the reported distribution of subtype B virus. In addition, multinucleated giant cells were significantly more common in HIV infected nodes (64%) compared to controls (4%) (P=0.00002). In conclusion, Langerhans histiocytes and related cells are reservoirs for HIV subtype E in lymph nodes. Disrupting the pathway of infection of Langerhans cells and related cells may be a viable strategy to interfere with transmission of HIV subtype E. PMID- 16341148 TI - Evidence for differential expression of Notch receptors and their ligands in melanocytic nevi and cutaneous malignant melanoma. AB - The Notch signaling has been implicated in the regulation of self-renewal of adult stem cells and differentiation of precursors along a specific cell lineage, in normal embryonic development and organogenesis. There is also evidence that signaling through Notch receptors regulate cell proliferation and cell survival in several types of cancer, with opposing results depending on tissue context. No data are available in the literature concerning modulation of the expression of Notch receptors, and their ligands, in human cutaneous malignant melanoma. Here, we have investigated, for the first time, the expression of Notch-1, Notch-2, Jagged-1, Jagged-2 and Delta-like 1 proteins, by immunohistochemistry, in a series of benign and malignant human melanocytic lesions: five common melanocytic nevi, five 'dysplastic nevi' and 20 melanomas (five in situ, five T1-T2, five T3 T4 and five metastatic melanomas). We found that the expression of Notch-1 and Notch-2, as well as Notch ligands, was upregulated in 'dysplastic nevi' and melanomas as compared with common melanocytic nevi. These results indicate that the activation of Notch may represent an early event in melanocytic tumor growth and upregulation of Notch signaling may sustain tumor progression. PMID- 16341149 TI - The Akt pathway in human breast cancer: a tissue-array-based analysis. AB - The Akt pathway, an important regulator of cell proliferation and survival, is deregulated in many cancers. The pathway has achieved considerable importance due to the development of kinase inhibitors that are able to successfully reduce tumor growth. This study was conducted to determine the status of the Akt pathway in human breast cancers and to study the relationship between the different component proteins. Expression levels of PTEN, phosphorylated forms of the constituent proteins (Akt, FKHR, mTOR, and S6) and cyclin D1 were evaluated by immunohistochemistry, on consecutive sections from a tissue microarray containing 145 invasive breast cancers and 140 pure ductal carcinomas in-situ. Aberrant expression was correlated statistically with tumor characteristics and disease outcome. The Akt pathway was found to be activated early in breast cancer, in the in-situ stage. In all, 33, 15, 32, and 60% of ductal carcinoma in-situ showed overexpression of Akt, FKHR, mTOR, and cyclin D1. PTEN loss did not correlate statistically with expression of AKT or any of the other proteins with the exception of S6, indicating that Akt activation was not a result of PTEN loss. Expression levels of PTEN and S6 were significantly different in in-situ and invasive cancers, indicating association with disease progression. Loss of PTEN was noted in 11% of in-situ as compared to 26% of invasive cancers, while S6 overexpression was seen in 47% in-situ and in 72% invasive cancers. High-grade carcinomas were associated with PTEN loss, while low-grade carcinomas with good prognostic features showed cyclin D1 overexpression and were associated with longer disease free survival. Additionally, cancers with mTOR overexpression showed a three times greater risk for disease recurrence. Overall, a large proportion of in-situ and invasive breast cancers overexpressed cyclinD1 and S6. Our results may have significant implications in the development and application of targeted therapy. PMID- 16341150 TI - Cardiac nonamyloidotic immunoglobulin deposition disease. AB - Cardiac nonamyloidotic immunoglobulin (Ig) deposition disease (CIDD) is a rare disorder characterized by Ig deposition in the myocardium associated with plasma cell dyscrasias. A retrospective review of cardiac biopsies performed at two different institutions identified eight patients with CIDD. All patients had plasma cell dyscrasias with monoclonal gammopathy. Three had IgG lambda, two had IgG kappa, one had IgD kappa and one each had free kappa and free lambda light chain. Four patients had concurrent amyloidosis involving other organs. One had amyloidosis of kidney alone, one had amyloidosis of kidney and abdominal fat pad and two others had amyloidosis of bone marrow vasculature. Three patients had dialysis-dependent renal insufficiency. None of the patients had symptoms of heart failure. Six patients had echocardiographically demonstrable concentric left ventricular hypertrophy with diastolic dysfunction. Two patients had significant cardiac arrhythmias requiring medical intervention. On endomyocardial biopsy, all eight had normal appearing myocardium on light microscopy with negative Congo Red and Thioflavin T stains. On immunofluorescent staining of the cardiac biopsies, all eight stained positive for interstitial Ig deposition. Electron microscopy (EM) confirmed the presence of granular deposits of Igs in the myocardium in five of the eight patients. EM studies were not available in one patient and two others had normal EM studies. In conclusion, CIDD should be considered in the spectrum of cardiovascular pathology in patients with plasma cell dyscrasias. They often, but not always, have left ventricular hypertrophy. These patients may be at risk for developing arrhythmias as well as diastolic dysfunction. Unless immunofluorescent and EM studies are performed routinely in biopsy material, this entity may be missed in the absence of amyloidosis. Concurrent amyloidosis in other organs sheds a unique perspective into the role of local microenvironment in the pathogenesis of systemic Ig deposition disease and amyloidosis. PMID- 16341151 TI - MALT1 and BCL10 aberrations in MALT lymphomas and their effect on the expression of BCL10 in the tumour cells. AB - Among the genetic abnormalities reported to occur in mucosa-associated lymphoid tissue (MALT) lymphomas, the three translocations t(11;18)(q21;q21), t(1;14)(p22;q32) and t(14;18)(q32;q21) are of particular interest because they appear to be specific for, or at least closely related to this type of B-cell non Hodgkin's lymphoma. These translocations affect the MALT1 (18q21) and BCL10 (1p22) genes. We retrieved 77 consecutive biopsies of MALT lymphomas (documented with frozen material) over a 10-year period and investigated these cases for the presence of these three translocations with fluorescence in situ hybridisation, along with the immunohistochemical analysis of the intracellular localisation of the BCL10 protein. The above-listed translocations occurred mutually exclusive and were detected in 10, 1 and 3% of the cases, respectively (the latter incidence being much lower than in the previously reported studies by one single group). These genetic rearrangements corresponded well with the aberrant subcellular localisation of the BCL10 protein as found by immunohistochemistry: t(11;18)(q21;q21) and (1;14)(p22;q32) were marked by a, respectively, moderate to strong nuclear BCL10 staining pattern while t(14;18)(q32;q21)-positive MALT lymphomas were characterised by a perinuclear BCL10 staining pattern. This study further supports the close interaction between the MALT1 and BCL10 proteins in the pathogenesis of MALT lymphomas and may indicate that BCL10 immunohistochemistry is a simple technique to identify those MALT lymphoma cases with an underlying genetic aberration. PMID- 16341152 TI - Current practice of diagnosis and reporting of prostatic intraepithelial neoplasia and glandular atypia among genitourinary pathologists. AB - The criteria for diagnosing prostatic intraepithelial neoplasia (PIN) and lesions suspicious for cancer are described in the literature. However, it is unknown how these are applied in practice by experts in genitourinary (GU) pathology. A questionnaire was sent to 93 GU pathologists in countries around the world with the purpose of surveying current practices. The response rate was 69% including 40 North American pathologists and 24 from other continents. For preneoplastic lesions, the term PIN was universally endorsed by the respondents. PIN was graded by 83%, usually as low/high-grade PIN (LGPIN/HGPIN) or as HGPIN only. Most respondents would usually not report lesions that may qualify for LGPIN. A majority (81%) did not specify architectural patterns of PIN. With both HGPIN and invasive cancer present, 69% would still mention HGPIN. Among the diagnostic criteria for HGPIN were any nucleoli visible (52%), or nucleoli seen in at least 10% of cells (33%). However, 56% would diagnose HGPIN in the absence of prominent nucleoli, most commonly based on prominent pleomorphism, marked hyperchromasia or mitotic figures. The number of cores involved with HGPIN was specified by 50%. Lesions suspicious for but not diagnostic of carcinoma were reported by 45% as atypia, atypical glands or suspicious for cancer and by 42% as atypical small acinar proliferation. The degree of suspicion was further defined by 41%. Our survey data may serve as a guideline to general pathologists on how to diagnose and report atypia and PIN in prostate biopsies. PMID- 16341153 TI - Interobserver agreement and reproducibility in classification of invasive breast carcinoma: an NCI breast cancer family registry study. AB - The United States National Cancer Institute Breast/Ovarian Cancer Family Registry is the largest international Registry of this type; over 37 724 individuals have been enrolled to date. One activity of this Registry is the semicentralized pathologic review of tumors from all probands. Given the semicentralized nature of the review, this study was undertaken to determine the reproducibility, source(s) of classification discrepancies and stratagems to circumvent discrepancies for histologic subtyping and grading of invasive breast cancer among the reviewing pathologists. A total of 13 pathologists reviewed 35 invasive breast cancers and classified them by primary and secondary histologic type, Nottingham grade and score. Lymph-vascular space invasion, circumscribed margins, syncytial growth and lymphocytic infiltrate were also evaluated. A training session using a separate set of slides was conducted prior to the study. General agreement, in terms of category-specific kappa's and percent agreement, and accuracy of classification relative to a reference standard were determined. Classification of histologic subtype was most consistent (and accurate) for mucinous carcinoma (kappa=1.0), followed by tubular (kappa=0.8) and lobular subtypes (kappa=0.8). Classification of medullary subtype was moderate (kappa=0.4), but additional evaluation of degree of lymphocytic infiltrate, syncytial growth and circumscribed margins identified most cases. Category specific kappa's were moderate to good for Nottingham grade (kappa=0.5-0.7), with the greatest agreement obtained in categorizing grade I (kappa=0.7), and grade III tumors (kappa=0.7). A flexible classification strategy that employs individual and combined criteria provides good interobserver agreement for invasive breast cancers with uniform, unambiguous histology and compensates for classification discrepancies in the more histologically ambiguous or heterogeneous cancers. PMID- 16341154 TI - CPD revisited. PMID- 16341155 TI - Glucose advice. PMID- 16341156 TI - NHS:private ratio. PMID- 16341157 TI - Crown distress. PMID- 16341158 TI - A word of thanks. PMID- 16341159 TI - Tennis ball treatment. PMID- 16341160 TI - Out-of-hours dental care. PMID- 16341162 TI - Periodontal therapy and pregnancy. PMID- 16341166 TI - Cranberries fight bacteria. PMID- 16341167 TI - Scottish children brush away tooth decay. PMID- 16341176 TI - One drug tackles two diseases. PMID- 16341177 TI - Oral medicine -- update for the dental practitioner. Disorders of orofacial sensation and movement. AB - This series provides an overview of current thinking in the more relevant areas of oral medicine for primary care practitioners, written by the authors while they were holding the Presidencies of the European Association for Oral Medicine and the British Society for Oral Medicine, respectively. A book containing additional material will be published. The series gives the detail necessary to assist the primary dental clinical team caring for patients with oral complaints that may be seen in general dental practice. Space precludes inclusion of illustrations of uncommon or rare disorders, or discussion of disorders affecting the hard tissues. Approaching the subject mainly by the symptomatic approach -- as it largely relates to the presenting complaint -- was considered to be a more helpful approach for GDPs rather than taking a diagnostic category approach. The clinical aspects of the relevant disorders are discussed, including a brief overview of the aetiology, detail on the clinical features and how the diagnosis is made. Guidance on management and when to refer is also provided, along with relevant websites which offer further detail. PMID- 16341178 TI - Content analysis of children's television advertising in relation to dental health. AB - This paper provides some disturbing facts and figures about the amount of television being watched by children. In addition, it reports on the volume and type of television advertising aimed at young people, both in the United Kingdom and other developed countries. In view of recent public and professional concern as to the possible adverse effects of food advertising on children's health, this study set out to examine what proportion of television advertisements, directed at children, promoted products potentially harmful to dental health. Forty-one hours of children's television programming broadcast on ITV1, the main UK commercial channel, were recorded on to videotape for subsequent analysis. Almost 1,000 adverts were analysed; each was timed and broadly categorised as relating to a food/drink product or non food/drink product. Advertisements for food and drink were further subdivided according to their sugar and/or acid content. We found that, on average, 24 adverts were shown per broadcast hour, which accounted for 15.8% of the total schedule time. 34.8% of adverts related to food/drink products, and 95.3% of these promoted products that were deemed potentially cariogenic or erosive. The most frequently promoted food/drink products included breakfast cereals with added sugar (26.3%), confectionery (23.7%) and non carbonated soft drinks (18.1%). It is very concerning that, despite recent specific codes of practice outlined by the Independent Television Commission for Children's Advertising, many food and drink products promoted during children's programming are potentially damaging to dental health. PMID- 16341179 TI - A review of prosthodontic management of fibrous ridges. AB - 'Fibrous' or 'flabby' alveolar ridges pose significant problems for the provision of stable and retentive dental prostheses for affected patients. In particular, problems arise during the act of impression taking, when forces cause the mobile denture bearing tissues to become distorted. The purpose of this paper is to review the impression techniques that can be used to optimise the treatment of edentulous patients with 'flabby' alveolar ridges. PMID- 16341186 TI - Teledentistry for screening new patient orthodontic referrals. Part 2: GDP perception of the referral system. AB - OBJECTIVE: To evaluate GDP opinion about a teledentistry system to screen new patient orthodontic referrals. STUDY DESIGN: Cross-sectional questionnaire. SAMPLE: Two hundred general dental practitioners (GDPs) were approached from Stockport, Rochdale, Oldham, Bury and Bolton in Greater Manchester, and High Peak in Derbyshire. METHOD: A questionnaire about a teledentistry system for new orthodontic patients was developed, following interviews with eight GDPs participating in a teledentistry trial. The questionnaire was posted to another 200 GDPs who were asked questions on issues such as the efficiency of a teledentistry system; the usefulness of a teledentistry system from the patients' point of view; their view of a teledentistry system; and any concerns they had relating to security, confidentiality and consent. RESULTS: Seventy one per cent of GDPs thought teledentistry for orthodontic referrals would be a good idea. At least 90% of responders agreed or neither agreed nor disagreed that patients would benefit from such a system. Over half of GDPs agreed or strongly agreed that there would be implications on their surgery time, expense and equipment security. CONCLUSIONS: GDPs generally supported a teledentistry system for new patient orthodontic referrals. Although perceived patient advantages were agreed, GDPs tended to be less sure about the impact on them in terms of set-up expenses, time in the surgery and appropriate remuneration. PMID- 16341187 TI - Poor patient awareness of the relationship between smoking and periodontal diseases. AB - OBJECTIVE: To investigate patients' knowledge of the effects of smoking on periodontal health. DESIGN: Patient answered questionnaires, which were anonymous. SETTING: Patients who attended GKT Dental Institute, King's College, London for dental treatment. SUBJECTS: One thousand patients attending Restorative Consultant Clinics and Primary Dental Care. RESULTS: Seventy-eight per cent of patients were aware that smoking had a negative impact on health. However, 52% of these patients who were aware could not state what the negative effects were on oral health. Seven per cent of patients stated that smoking affected the gums but did not state how it affected the gums. Only 6% of respondents knew specifically of the link between smoking and periodontal disease. The only statistically significant factors associated with increased awareness were smoking status, ie being a non-smoker and being registered with a general dental practitioner. Non-smokers and those registered with GDPs were more likely to be aware of the association between smoking and periodontal diseases. CONCLUSIONS: This study highlights patients' lack of awareness of the relationship between smoking and periodontal diseases, with only 6% of respondents knowing of the link between tobacco and periodontal diseases. PMID- 16341197 TI - 125 years of developments in dentistry, 1880-2005. Part 6: General and specialist practice. PMID- 16341202 TI - Facioscapulohumeral muscular dystrophy in mice overexpressing FRG1. AB - Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that is not due to a classical mutation within a protein coding gene. Instead, almost all FSHD patients carry deletions of an integral number of tandem 3.3-kilobase repeat units, termed D4Z4, located on chromosome 4q35 (ref. 3). D4Z4 contains a transcriptional silencer whose deletion leads to inappropriate overexpression in FSHD skeletal muscle of 4q35 genes located upstream of D4Z4 (ref. 4). To identify the gene responsible for FSHD pathogenesis, we generated transgenic mice selectively overexpressing in skeletal muscle the 4q35 genes FRG1, FRG2 or ANT1. We find that FRG1 transgenic mice develop a muscular dystrophy with features characteristic of the human disease; by contrast, FRG2 and ANT1 transgenic mice seem normal. FRG1 is a nuclear protein and several lines of evidence suggest it is involved in pre-messenger RNA splicing. We find that in muscle of FRG1 transgenic mice and FSHD patients, specific pre-mRNAs undergo aberrant alternative splicing. Collectively, our results suggest that FSHD results from inappropriate overexpression of FRG1 in skeletal muscle, which leads to abnormal alternative splicing of specific pre mRNAs. PMID- 16341203 TI - GABA regulates synaptic integration of newly generated neurons in the adult brain. AB - Adult neurogenesis, the birth and integration of new neurons from adult neural stem cells, is a striking form of structural plasticity and highlights the regenerative capacity of the adult mammalian brain. Accumulating evidence suggests that neuronal activity regulates adult neurogenesis and that new neurons contribute to specific brain functions. The mechanism that regulates the integration of newly generated neurons into the pre-existing functional circuitry in the adult brain is unknown. Here we show that newborn granule cells in the dentate gyrus of the adult hippocampus are tonically activated by ambient GABA (gamma-aminobutyric acid) before being sequentially innervated by GABA- and glutamate-mediated synaptic inputs. GABA, the major inhibitory neurotransmitter in the adult brain, initially exerts an excitatory action on newborn neurons owing to their high cytoplasmic chloride ion content. Conversion of GABA-induced depolarization (excitation) into hyperpolarization (inhibition) in newborn neurons leads to marked defects in their synapse formation and dendritic development in vivo. Our study identifies an essential role for GABA in the synaptic integration of newly generated neurons in the adult brain, and suggests an unexpected mechanism for activity-dependent regulation of adult neurogenesis, in which newborn neurons may sense neuronal network activity through tonic and phasic GABA activation. PMID- 16341204 TI - Antiviral treatment is more effective than smallpox vaccination upon lethal monkeypox virus infection. AB - There is concern that variola virus, the aetiological agent of smallpox, may be used as a biological weapon. For this reason several countries are now stockpiling (vaccinia virus-based) smallpox vaccine. Although the preventive use of smallpox vaccination has been well documented, little is known about its efficacy when used after exposure to the virus. Here we compare the effectiveness of (1) post-exposure smallpox vaccination and (2) antiviral treatment with either cidofovir (also called HPMPC or Vistide) or with a related acyclic nucleoside phosphonate analogue (HPMPO-DAPy) after lethal intratracheal infection of cynomolgus monkeys (Macaca fascicularis) with monkeypox virus (MPXV). MPXV causes a disease similar to human smallpox and this animal model can be used to measure differences in the protective efficacies of classical and new-generation candidate smallpox vaccines. We show that initiation of antiviral treatment 24 h after lethal intratracheal MPXV infection, using either of the antiviral agents and applying various systemic treatment regimens, resulted in significantly reduced mortality and reduced numbers of cutaneous monkeypox lesions. In contrast, when monkeys were vaccinated 24 h after MPXV infection, using a standard human dose of a currently recommended smallpox vaccine (Elstree-RIVM), no significant reduction in mortality was observed. When antiviral therapy was terminated 13 days after infection, all surviving animals had virus-specific serum antibodies and antiviral T lymphocytes. These data show that adequate preparedness for a biological threat involving smallpox should include the possibility of treating exposed individuals with antiviral compounds such as cidofovir or other selective anti-poxvirus drugs. PMID- 16341205 TI - Histone acetylation by Trrap-Tip60 modulates loading of repair proteins and repair of DNA double-strand breaks. AB - DNA is packaged into chromatin, a highly compacted DNA-protein complex; therefore, all cellular processes that use the DNA as a template, including DNA repair, require a high degree of coordination between the DNA-repair machinery and chromatin modification/remodelling, which regulates the accessibility of DNA in chromatin. Recent studies have implicated histone acetyltransferase (HAT) complexes and chromatin acetylation in DNA repair; however, the precise underlying mechanism remains poorly understood. Here, we show that the HAT cofactor Trrap and Tip60 HAT bind to the chromatin surrounding sites of DNA double-strand breaks (DSBs) in vivo. Trrap depletion impairs both DNA-damage induced histone H4 hyperacetylation and accumulation of repair molecules at sites of DSBs, resulting in defective homologous recombination (HR) repair, albeit with the presence of a functional ATM-dependent DNA-damage signalling cascade. Importantly, the impaired loading of repair proteins and the defect in DNA repair in Trrap-deficient cells can be counteracted by chromatin relaxation, indicating that the DNA-repair defect that was observed in the absence of Trrap is due to impeded chromatin accessibility at sites of DNA breaks. Thus, these data reveal that cells may use the same basic mechanism involving HAT complexes to regulate distinct cellular processes, such as transcription and DNA repair. PMID- 16341206 TI - Flotillin-1 defines a clathrin-independent endocytic pathway in mammalian cells. AB - Previous studies provide evidence for an endocytic mechanism in mammalian cells that is distinct from both clathrin-coated pits and caveolae, and is not inhibited by overexpression of GTPase-null dynamin mutants. This mechanism, however, has been defined largely in these negative terms. We applied a ferro fluid-based purification of endosomes to identify endosomal proteins. One of the proteins identified in this way was flotillin-1 (also called reggie-2). Here, we show that flotillin-1 resides in punctate structures within the plasma membrane and in a specific population of endocytic intermediates. These intermediates accumulate both glycosylphosphatidylinositol (GPI)-linked proteins and cholera toxin B subunit. Endocytosis in flotillin-1-containing intermediates is clathrin independent. Total internal reflection microscopy and immuno-electron microscopy revealed that flotillin-1-containing regions of the plasma membrane seem to bud into the cell, and are distinct from clathrin-coated pits and caveolin-1-positive caveolae. Flotillin-1 small interfering RNA (siRNA) inhibited both clathrin independent uptake of cholera toxin and endocytosis of a GPI-linked protein. We propose that flotillin-1 is one determinant of a clathrin-independent endocytic pathway in mammalian cells. PMID- 16341207 TI - The tumour-suppressor genes NF2/Merlin and Expanded act through Hippo signalling to regulate cell proliferation and apoptosis. AB - Merlin, the protein product of the Neurofibromatosis type-2 gene, acts as a tumour suppressor in mice and humans. Merlin is an adaptor protein with a FERM domain and it is thought to transduce a growth-regulatory signal. However, the pathway through which Merlin acts as a tumour suppressor is poorly understood. Merlin, and its function as a negative regulator of growth, is conserved in Drosophila, where it functions with Expanded, a related FERM domain protein. Here, we show that Drosophila Merlin and Expanded are components of the Hippo signalling pathway, an emerging tumour-suppressor pathway. We find that Merlin and Expanded, similar to other components of the Hippo pathway, are required for proliferation arrest and apoptosis in developing imaginal discs. Our genetic and biochemical data place Merlin and Expanded upstream of Hippo and identify a pathway through which they act as tumour-suppressor genes. PMID- 16341208 TI - Cdk5 phosphorylates and stabilizes p27kip1 contributing to actin organization and cortical neuronal migration. AB - p27(kip1), a cyclin-dependent kinase (CDK) inhibitor (CKI), generally suppresses CDK activity in proliferating cells. Although another role of p27 in cell migration has been recently suggested in vitro, the physiological importance of p27 in cell migration remains elusive, as p27-deficient mice have not shown any obvious migration-defect-related phenotypes. Here, we show that Cdk5, an unconventional neuronal CDK, phosphorylates and stabilizes p27 as an upstream regulator, maintaining the amount of p27 in post-mitotic neurons. In vivo RNA interference (RNAi) experiments showed that reduced amounts of p27 caused inhibition of cortical neuronal migration and decreased the amount of F-actin in the processes of migrating neurons. The Cdk5-p27 pathway activates an actin binding protein, cofilin, which is also shown to be involved in cortical neuronal migration in vivo. Our findings shed light on a previously unknown new relationship between CDK and CKI in G0-arrested cells that regulates cytoskeletal reorganization and neuronal migration during corticogenesis. PMID- 16341209 TI - Inactivation of the central thalamus delays self-timed saccades. AB - The central thalamus transmits corollary discharge signals for eye movement control, but its role in eye movement generation remains uncertain. Inactivation of the paralaminar part of the ventrolateral thalamus delayed the initiation of contraversive saccades, particularly during a new memory-guided saccade task that required self-triggering of the movement. The results suggest that signals through the thalamus regulate the timing of self-initiated saccades. PMID- 16341210 TI - Coherent spontaneous activity accounts for trial-to-trial variability in human evoked brain responses. AB - Trial-to-trial variability in the blood oxygen level-dependent (BOLD) response of functional magnetic resonance imaging has been shown to be relevant to human perception and behavior, but the sources of this variability remain unknown. We demonstrate that coherent spontaneous fluctuations in human brain activity account for a significant fraction of the variability in measured event-related BOLD responses and that spontaneous and task-related activity are linearly superimposed in the human brain. PMID- 16341211 TI - Axons and dendrites originate from neuroepithelial-like processes of retinal bipolar cells. AB - The cellular mechanisms underlying axogenesis and dendritogenesis are not completely understood. The axons and dendrites of retinal bipolar cells, which contact their synaptic partners within specific laminae in the inner and outer retina, provide a good system for exploring these issues. Using transgenic mice expressing enhanced green fluorescent protein (GFP) in a subset of bipolar cells, we determined that axonal and dendritic arbors of these interneurons develop directly from apical and basal processes attached to the outer and inner limiting membranes, respectively. Selective stabilization of processes contributed to stratification of axonal and dendritic arbors within the appropriate synaptic layer. This unusual mode of axogenesis and dendritogenesis from neuroepithelial like processes may act to preserve neighbor-neighbor relationships in synaptic wiring between the outer and inner retina. PMID- 16341212 TI - ROBO directs axon crossing of segmental boundaries by suppressing responsiveness to relocalized Netrin. AB - Networks in the CNS consist of neural modules that are connected in a repetitive array. Whereas individual modules contain guidance information along which axons track within the unit, these guidance cues hinder axon extension across module boundaries. We investigated how axons solve this 'boundary problem' by analyzing the longitudinal connections of neuromeres in Drosophila melanogaster. The initial trajectory of the longitudinal axons is guided by Netrin, which is localized on commissural axons by its receptor, Frazzled. The Netrin cue on the commissure of the next segment can act as a barrier to longitudinal axons, inhibiting their extension and misguiding them contralaterally along the commissure. We show that, before reaching the segmental boundary, the longitudinal axons' responsiveness to Netrin presented on the commissure is suppressed by Roundabout (ROBO), through counteracting Gq signaling. The absence of suppression causes the robo phenotype: longitudinal axons project toward the midline, as if running around a roundabout (rotary). PMID- 16341214 TI - Development of shared information in communication despite hippocampal amnesia. AB - This study investigated the ability of individuals with amnesia to acquire referential labels across a series of collaborative, dynamic interactions with a communication partner. Despite their inability to learn arbitrary relations in paired-associate learning, the amnesic patients showed remarkably robust collaborative learning across trials, at a rate equal to that of normal comparison subjects. The amnesic participants' learning resulted in their arriving at labels for a set of abstract shapes (tangrams), thus facilitating rapid and efficient communication. The labels generated and used by the amnesics during interactions with their partners became simpler across trials, with most labels stabilizing long before the end of training and then being used consistently throughout; moreover, they endured long after the task had ended. These findings have important implications for understanding the memory systems involved in semantic learning and in acquiring shared knowledge ('common ground') among communication partners, and the nature of hippocampal-dependent versus hippocampal-independent learning. PMID- 16341213 TI - CaM kinase II phosphorylation of slo Thr107 regulates activity and ethanol responses of BK channels. AB - High-conductance, Ca(2+)-activated and voltage-gated (BK) channels set neuronal firing. They are almost universally activated by alcohol, leading to reduced neuronal excitability and neuropeptide release and to motor intoxication. However, several BK channels are inhibited by alcohol, and most other voltage gated K(+) channels are refractory to drug action. BK channels are homotetramers (encoded by Slo1) that possess a unique transmembrane segment (S0), leading to a cytosolic S0-S1 loop. We identified Thr107 of bovine slo (bslo) in this loop as a critical residue that determines BK channel responses to alcohol. In addition, the activity of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) in the cell controlled channel activity and alcohol modulation. Incremental CaMKII mediated phosphorylation of Thr107 in the BK tetramer progressively increased channel activity and gradually switched the channel alcohol responses from robust activation to inhibition. Thus, CaMKII phosphorylation of slo Thr107 works as a 'molecular dimmer switch' that could mediate tolerance to alcohol, a form of neuronal plasticity. PMID- 16341215 TI - The claw paw mutation reveals a role for Lgi4 in peripheral nerve development. AB - Peripheral nerve development results from multiple cellular interactions between axons, Schwann cells and the surrounding mesenchymal tissue. The delayed axonal sorting and hypomyelination throughout the peripheral nervous system of claw paw (clp) mutant mice suggest that the clp gene product is critical for these interactions. Here we identify the clp mutation as a 225-bp insertion in the Lgi4 gene. Lgi4 encodes a secreted and glycosylated leucine-rich repeat protein and is expressed in Schwann cells. The clp mutation affects Lgi4 mRNA splicing, resulting in a mutant protein that is retained in the cell. Additionally, siRNA mediated downregulation of Lgi4 in wild-type neuron-Schwann cell cocultures inhibits myelination, whereas exogenous Lgi4 restores myelination in clp/clp cultures. Thus, the abnormalities observed in clp mice are attributable to the loss of Lgi4 function, and they identify Lgi4 as a new component of Schwann cell signaling pathway(s) that controls axon segregation and myelin formation. PMID- 16341216 TI - The thymus exports long-lived fully committed T cell precursors that can colonize primary lymphoid organs. AB - Thymic export of cells is believed to be restricted to mature T cells. Here we show that the thymus also exports fully committed T cell precursors that colonize primary lymphoid organs. These precursor cells exited the thymus before T cell receptor rearrangements and colonized lymphoid organs such as the thymus and the gut. Migration of the thymic T cell-committed precursors led to permanent colonization of the gut precursor compartment, improved the capacity of gut precursors to further differentiate into T cells and was sufficient for the generation of 'euthymic like' CD8alphaalpha(+) intraepithelial lymphocytes. These data demonstrate a new function for the thymus in peripheral seeding with T cell precursors that become long lived after thymus export. PMID- 16341217 TI - Intracellular localization of Toll-like receptor 9 prevents recognition of self DNA but facilitates access to viral DNA. AB - Toll-like receptors (TLRs) sense infection by detecting molecular structures of microbial origin. TLR3, TLR7 and TLR9 recognize nucleic acids and are localized to intracellular compartments where they normally respond to viral nucleic acids. The purpose for this intracellular localization, however, is not clear. Here we describe a chimeric TLR9 receptor that localized to the cell surface and responded normally to synthetic TLR9 ligands but not to viral nucleic acids. However, the 'relocated' chimeric TLR9 receptor was able to recognize self DNA, which does not stimulate wild-type TLR9. These data demonstrated that intracellular localization of TLR9 was not required for ligand recognition. Instead, localization of the nucleic acid-sensing TLRs is critical in discriminating between self and nonself nucleic acid. PMID- 16341218 TI - From brittle to ductile fracture of bone. AB - Toughness is crucial to the structural function of bone. Usually, the toughness of a material is not just determined by its composition, but by the ability of its microstructure to dissipate deformation energy without propagation of the crack. Polymers are often able to dissipate energy by viscoplastic flow or the formation of non-connected microcracks. In ceramics, well-known toughening mechanisms are based on crack ligament bridging and crack deflection. Interestingly, all these phenomena were identified in bone, which is a composite of a fibrous polymer (collagen) and ceramic nanoparticles (carbonated hydroxyapatite). Here, we use controlled crack-extension experiments to explain the influence of fibre orientation on steering the various toughening mechanisms. We find that the fracture energy changes by two orders of magnitude depending on the collagen orientation, and the angle between collagen and crack propagation direction is decisive in switching between different toughening mechanisms. PMID- 16341221 TI - Dosage compensation of the active X chromosome in mammals. AB - Monosomy of the X chromosome owing to divergence between the sex chromosomes leads to dosage compensation mechanisms to restore balanced expression between the X and the autosomes. In Drosophila melanogaster, upregulation of the male X leads to dosage compensation. It has been hypothesized that mammals likewise upregulate their active X chromosome. Together with X inactivation, this mechanism would maintain balanced expression between the X chromosome and autosomes and between the sexes. Here, we show that doubling of the global expression level of the X chromosome leads to dosage compensation in somatic tissues from several mammalian species. X-linked genes are highly expressed in brain tissues, consistent with a role in cognitive functions. Furthermore, the X chromosome is expressed but not upregulated in spermatids and secondary oocytes, preserving balanced expression of the genome in these haploid cells. Upon fertilization, upregulation of the active X must occur to achieve the observed dosage compensation in early embryos. PMID- 16341222 TI - Defective planar cell polarity in polycystic kidney disease. AB - Morphogenesis involves coordinated proliferation, differentiation and spatial distribution of cells. We show that lengthening of renal tubules is associated with mitotic orientation of cells along the tubule axis, demonstrating intrinsic planar cell polarization, and we demonstrate that mitotic orientations are significantly distorted in rodent polycystic kidney models. These results suggest that oriented cell division dictates the maintenance of constant tubule diameter during tubular lengthening and that defects in this process trigger renal tubular enlargement and cyst formation. PMID- 16341223 TI - Genetic analysis of cavefish reveals molecular convergence in the evolution of albinism. AB - The genetic basis of vertebrate morphological evolution has traditionally been very difficult to examine in naturally occurring populations. Here we describe the generation of a genome-wide linkage map to allow quantitative trait analysis of evolutionarily derived morphologies in the Mexican cave tetra, a species that has, in a series of independent caves, repeatedly evolved specialized characteristics adapted to a unique and well-studied ecological environment. We focused on the trait of albinism and discovered that it is linked to Oca2, a known pigmentation gene, in two cave populations. We found different deletions in Oca2 in each population and, using a cell-based assay, showed that both cause loss of function of the corresponding protein, OCA2. Thus, the two cave populations evolved albinism independently, through similar mutational events. PMID- 16341224 TI - Deletion of Peg10, an imprinted gene acquired from a retrotransposon, causes early embryonic lethality. AB - By comparing mammalian genomes, we and others have identified actively transcribed Ty3/gypsy retrotransposon-derived genes with highly conserved DNA sequences and insertion sites. To elucidate the functions of evolutionarily conserved retrotransposon-derived genes in mammalian development, we produced mice that lack one of these genes, Peg10 (paternally expressed 10), which is a paternally expressed imprinted gene on mouse proximal chromosome 6. The Peg10 knockout mice showed early embryonic lethality owing to defects in the placenta. This indicates that Peg10 is critical for mouse parthenogenetic development and provides the first direct evidence of an essential role of an evolutionarily conserved retrotransposon-derived gene in mammalian development. PMID- 16341225 TI - Crystal structure and functional analysis of Dcp2p from Schizosaccharomyces pombe. AB - Decapping is a key step in both general and nonsense-mediated 5' --> 3' mRNA decay pathways. Removal of the cap structure is catalyzed by the Dcp1-Dcp2 complex. The crystal structure of a C-terminally truncated Schizosaccharomyces pombe Dcp2p reveals two distinct domains: an all-helical N-terminal domain and a C-terminal domain that is a classic Nudix fold. The C-terminal domain of both Saccharomyces cerevisiae and S. pombe Dcp2p proteins is sufficient for decapping activity, although the N-terminal domain can affect the efficiency of Dcp2p function. The binding of Dcp2p to Dcp1p is mediated by a conserved surface on its N-terminal domain, and the N-terminal domain is required for Dcp1p to stimulate Dcp2p activity. The flexible nature of the N-terminal domain relative to the C terminal domain suggests that Dcp1p binding to Dcp2p may regulate Dcp2p activity through conformational changes of the two domains. PMID- 16341226 TI - Structure of an RNA polymerase II-RNA inhibitor complex elucidates transcription regulation by noncoding RNAs. AB - The noncoding RNA B2 and the RNA aptamer FC bind RNA polymerase (Pol) II and inhibit messenger RNA transcription initiation, but not elongation. We report the crystal structure of FC(*), the central part of FC RNA, bound to Pol II. FC(*) RNA forms a double stem-loop structure in the Pol II active center cleft. B2 RNA may bind similarly, as it competes with FC(*) RNA for Pol II interaction. Both RNA inhibitors apparently prevent the downstream DNA duplex and the template single strand from entering the cleft after DNA melting and thus interfere with open-complex formation. Elongation is not inhibited, as nucleic acids prebound in the cleft would exclude the RNA inhibitors. The structure also indicates that A form RNA could interact with Pol II similarly to a B-form DNA promoter, as suggested for the bacterial transcription inhibitor 6S RNA. PMID- 16341227 TI - An mRNA-rRNA base-pairing mechanism for translation initiation in eukaryotes. AB - Base-pairing of messenger RNA to ribosomal RNA is a mechanism of translation initiation in prokaryotes. Although analogous base-pairing has been suggested to affect the translation of various eukaryotic mRNAs, direct evidence has been lacking. To test such base-pairing, we developed a yeast system that uses ribosomes containing a mouse-yeast hybrid 18S rRNA. Using this system, we demonstrate that a 9-nucleotide element found in the mouse Gtx homeodomain mRNA facilitates translation initiation by base-pairing to 18S rRNA. Various point mutations in the Gtx element and in either the hybrid or wild-type yeast 18S rRNAs confirmed the requirement for an intact complementary match. The presence of the Gtx element in various mRNAs suggests that this element affects the translation of groups of mRNAs. We discuss the possibility that other mRNA elements affect translation by base-pairing to different sites in the 18S rRNA. PMID- 16341228 TI - The human SWI/SNF subunit Brm is a regulator of alternative splicing. AB - The SWI/SNF (mating-type switch/sucrose nonfermenting) complex involved in chromatin remodeling on promoters has also been detected on the coding region of genes. Here we show that SWI/SNF can function as a regulator of alternative splicing. We found that the catalytic subunit Brm favors inclusion of variant exons in the mRNA of several genes, including E-cadherin, BIM, cyclin D1 and CD44. Consistent with this, Brm associates with several components of the spliceosome and with Sam68, an ERK-activated enhancer of variant exon inclusion. Examination of the CD44 gene revealed that Brm induced accumulation of RNA polymerase II (RNAPII) with a modified CTD phosphorylation pattern on regions encoding variant exons. Altogether, our data suggest that on genes regulated by SWI/SNF, Brm contributes to the crosstalk between transcription and RNA processing by decreasing RNAPII elongation rate and facilitating recruitment of the splicing machinery to variant exons with suboptimal splice sites. PMID- 16341229 TI - Vincristine is an effective therapeutic approach for transplantation-associated thrombotic microangiopathy. PMID- 16341231 TI - Adenosine- and hypoxia-induced dilation of human coronary resistance arteries: evidence against the involvement of K(ATP) channels. AB - 1. The ATP-sensitive potassium (K(ATP)) channel may be an important mediator of metabolic dilation in the human coronary circulation. As adenosine and hypoxia are considered to be major mediators of metabolic dilation in the coronary circulation, we investigated the effect of glibenclamide (a K(ATP) channel blocker) on adenosine and hypoxic dilation of human coronary resistance arteries, with myogenic tone, in vitro. 2. Vessels were dissected from the atrial appendage from consenting patients and studied in vitro using a pressure arteriograph system. Segments of coronary resistance artery were pressurized to 60 mmHg and the vessels studied developed spontaneous myogenic tone. 3. The K(ATP) opener pinacidil (final conc. 5 x 10(-6) M) resulted in dilation, which was completely reversed by 5 x 10(-6) glibenclamide (84+/-14 vs -10+/-9%, pinacidil and pinacidil plus glibenclamide, respectively, P=0.009, n=5). 4. Adenosine (final conc. 10(-5) M) resulted in dilation, glibenclamide (5 x 10(-6) and 10(-5) M) was without effect (118+/-12 vs 104+/-16% adenosine and adenosine plus 10(-5) glibenclamide, respectively, n.s., n=4). 5. Hypoxia (8+/-3 mmHg O2) resulted in a dilation that reversed when normoxic conditions were restored (60+/-9 vs 3+/-11% hypoxia and post-hypoxia, respectively, P=0.014, n=3). The hypoxic dilation was not affected by glibenclamide (63+/-14 vs 55+/-6% hypoxia and hypoxia plus glibenclamide, respectively, n.s., n=4). In a further series of experiments, vessels were incubated with glibenclamide (5 x 10(-6)) prior to a hypoxic challenge; again, glibenclamide was without effect on the hypoxic dilation ( 0.008+/-2 vs 95+/-3% glibenclamide and glibenclamide plus hypoxia, respectively, P=0.0005, n=3). 6. These data demonstrate that glibenclamide is without effect on both adenosine and hypoxic dilation of human coronary resistance arteries with myogenic tone, from the right atrial appendage in vitro. Our findings suggest that the K(ATP) channel is unlikely to be a major mediator of metabolic dilation in these arteries. PMID- 16341232 TI - Altered responsiveness of the guinea-pig isolated ileum to smooth muscle stimulants and to electrical stimulation after in situ ischemia. AB - 1. We evaluated changes in contractility of the guinea-pig isolated ileum, using intact segments and myenteric plexus-longitudinal muscle (MPLM) preparations, after several times (5-160 min) of ischemia in situ. 2. Intestinal ischemia was produced by clamping the superior mesenteric artery. Ischemic and nonischemic segments, obtained from the same guinea-pig, were mounted in organ baths containing Krebs-bicarbonate (K-B) solution, maintained at 37 degrees C and gassed with 95% O2/5% CO2. The preparations were allowed to equilibrate for 60 min under continuous superfusion of warm K-B solution and then electrically stimulated at 40 V (0.3 Hz, 3.0 ms). Thereafter, complete noncumulative concentration-response curves were constructed for acetylcholine (ACh), histamine (HIS), potassium chloride (KCl), and barium chloride (BaCl2). Mean Emax (maximal response) values were calculated for each drug. 3. Our study shows that alterations of chemically and electrically evoked contractions are dependent on ischemic periods. It also demonstrates that contractile responses of ischemic tissues to neurogenic stimulation decreases earlier and to a significantly greater extent than the non-nerve mediated responses of the intestinal smooth muscle. Contractile responses to smooth muscle stimulants were all similarly affected by ischemia. Electron microscopy images indicated necrotic neuronal death. The decrease in reactivity of ischemic tissues to electrical stimulation was ameliorated by dexrazoxane, an antioxidant agent. 4. We consider the guinea pig isolated ileum as a useful model system to study the processes involved in neuronal ischemia, and we propose that the reduction in maximal responses to electrical stimulation is a useful parameter to study neuroprotection. PMID- 16341233 TI - Effects of the antifungal antibiotic clotrimazole on human cardiac repolarization potassium currents. AB - The antifungal antibiotic clotrimazole (CLT) shows therapeutic effects on cancer, sickle cell disease, malaria, etc. by inhibiting membrane intermediate conductance Ca2+ -activated K+ channels (IKCa). However, it is unclear whether this drug would affect human cardiac K+ currents. The present study was therefore designed to investigate the effects of CLT on transient outward K+ current (Ito1), and ultra-rapid delayed rectifier K+ current (IKur) in isolated human atrial myocytes, and cloned hERG channel current (IhERG) and recombinant human cardiac KCNQ1/KCNE1 channel current (IKs) expressed in HEK 293 cells. It was found that CLT inhibited Ito1 with an IC50 of 29.5 microM, accelerated Ito1 inactivation, and decreased recovery of Ito1 from inactivation. In addition, CLT inhibited human atrial I(Kur) in a concentration-dependent manner (IC50 = 7.6 microM). CLT substantially suppressed IhERG (IC50 = 3.6 microM), and negatively shifted the activation conductance of IhERG. Moreover, CLT inhibited IKs (IC50 = 15.1 microM), and positively shifted the activation conductance of the current. These results indicate that the antifungal antibiotic CLT substantially inhibits human cardiac repolarization K+ currents including Ito1, IKur, IhERG, and IKs. However, caution is recommended when correlating the observed in vitro effects on cardiac ion currents to the clinical relevance. PMID- 16341235 TI - Myelodysplastic syndromes (MDSs) and acute myelogenous leukemia (AML) comprise a closely linked continuum of malignant hematologic diseases. Introduction. PMID- 16341234 TI - Activation of ERK1/2 by extracellular nucleotides in macrophages is mediated by multiple P2 receptors independently of P2X7-associated pore or channel formation. AB - Macrophages express several P2X and P2Y nucleotide receptors and display the phenomenon of ATP-induced P2X7-dependent membrane permeabilization, which occurs through a poorly understood mechanism. Several P2 receptors are known to be coupled to the activation of mitogen-activated protein kinases (MAPKs) and Ca2+ signaling. Here, we use macrophages to investigate the phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) by nucleotides and the involvement of MAPKs and intracellular Ca2+ concentration in ATP-induced membrane permeabilization. Short-term (5 min) pre-exposure to oxidized ATP (oATP), a P2X7 antagonist that does not inhibit P2X7-associated inward currents or membrane permeabilization, inhibits the activation of ERK1/2 by ATP, ADP, the P2X7 agonist 2'-3'-O-(4-benzoylbenzoyl)-ATP (BzATP), but not by UTP and UDP. We conclude that macrophages display several P2Y receptors coupled to the ERK1/2 pathway and that oATP antagonizes the action of purine nucleotides, possibly binding to P2X7 and/or other purine-binding P2Y receptors. We also show that BzATP and ATP activate ERK1/2 by two different pathways since ERK1/2 activation by BzATP, but not by ATP, is blocked by the tryrosine kinase inhibitor, genistein, and the Src protein kinase inhibitor, tyrphostin. However, the activation of ERK1/2 by ATP is blocked by the protein kinase C (PKC) inhibitor, chelerythrine chloride. Under the same conditions, membrane permeabilization is not blocked by genistein, tyrphostin, or chelerythrine chloride, indicating that tyrosine kinase, Src protein kinase, and PKC are not required for pore opening. Membrane permeabilization is independent of ERK1/2 activation since chelerythrine, or short-term exposure to oATP or PD98059, efficiently block ERK1/2 activation without inhibiting membrane permeabilization. In addition, membrane permeabilization is not inhibited by SB203580 and SB202190, two inhibitors of p38 MAPK, nor by intracellular BAPTA, which blocks ATP-induced Ca2+ signals. These results suggest that multiple P2 receptors lead to ERK1/2 activation, that ligation of the same receptors by agonists with different affinities can lead to differential stimulation of separate pathways, and that MAPKs and intracellular Ca2+ fluxes are independent of P2X7-associated pore formation. PMID- 16341236 TI - Methylation inhibitor therapy in the treatment of myelodysplastic syndrome. AB - The class of DNA methyltransferase inhibitors is represented by azacitidine and decitabine. Azacitidine is approved for the treatment of patients in both low- and high-risk subtypes of myelodysplastic syndrome (MDS), and decitabine is currently under review by the FDA. Azacitidine phase III trial data, based upon the Cancer and Leukemia Group B (CALGB) study 9221, showed durable clinical and symptomatic improvement in bone marrow function, a reduction in the risk of leukemic transformation, and significant improvements in the quality of life of patients treated with azacitidine compared with supportive care alone. This study also provided data suggestive of improvement in survival in MDS patients. The experience with decitabine comprises a number of phase I/II studies and a phase III trial yet to be published. While there is a strong base of experience supporting the efficacy of DNA methyltransferase inhibitors in the treatment of MDS, a number of practical issues need to be explored further. These include the optimization of the timing and duration of treatment, and the prediction of response to therapy. Along with current experience, future studies will lead to the development of treatment algorithms, strategies for selecting patients (e.g. according to age, risk, classification, and cytogenetic profile), and the combination strategies, particularly with histone deacetylase inhibitors, in the management of MDS. PMID- 16341237 TI - Optimizing therapy with methylation inhibitors in myelodysplastic syndromes: dose, duration, and patient selection. AB - Azacitidine (Vidaza, Pharmion Corp., Boulder, CO, USA) and decitabine (Dacogentrade mark, SuperGen, Inc., Dublin, CA, USA, and MGI Pharma, Inc., Bloomington, MN, USA) are DNA methyltransferase (DNMT) inhibitors that have clinical activity in patients with myelodysplastic syndromes. Mechanism-based laboratory studies suggest that clinical optimization of therapy with DNMT inhibitors needs to include optimizing intracellular drug uptake and maximizing drug exposure over time while still using lower doses to avoid cytotoxicity. Clinical studies suggest that increased dose intensity and multiple cycles of administration substantially increase response rates. Other strategies for optimizing the efficacy of DNMT inhibitor therapy also include identification of patients that are best qualified for treatment, and defining in vivo mechanisms of patient responses. In the future, combination strategies to increase gene reactivation and to take advantage of increased expression of target genes may be critical for achieving optimal results. PMID- 16341238 TI - Combination therapy with DNA methyltransferase inhibitors in hematologic malignancies. AB - A variety of epigenetic changes contribute to transcriptional dysregulation in myelodysplastic syndromes (MDSs) and acute myeloid leukemia (AML). DNA methyltransferase (DNMT) inhibitors--azacitidine and decitabine--have significant activity in the treatment of MDS. Despite marked activity in myeloid malignancy, monotherapy with DNMT inhibitors is limited by low complete and partial response rates (7-20%) and median response durations of 15 months. As with classical cytotoxic therapy, the targeting of biologic pathways and mechanisms may best be accomplished using a combination of agents offering complementary mechanisms and synergistic pharmacodynamic interactions. The goal of this approach is to improve response rates, quality, and duration, and to minimize adverse events. There are a number of new therapies under development for the management of MDS and AML. This review article touches on some of the more promising combination regimens in various phases of investigation. The treatment of MDS and AML is undergoing rapid evolution. Cytogenetic complete remission and prolonged survival represent important goals. Incremental improvements in disease state and quality-of-life issues are also important for patients. Given the overall failure of cytotoxic chemotherapy in the achievement of cures in MDS and MDS-related AML, the application of less toxic, biologically directed agents may represent a more promising approach to treatment. Combination therapies with DNMT inhibitors using optimal dosing regimens to focus on methylation reversal with lower doses over longer periods of time, rather than direct cytotoxic effects, are beginning to suggest promising results in MDS and AML. PMID- 16341239 TI - Inhibitors of DNA methylation: beyond myelodysplastic syndromes. AB - DNA methyltransferase (DNMT) inhibitors, azacitidine (Vidaza, Pharmion, Boulder, CO, USA) and decitabine (Dacogen; SuperGen Inc, Dublin, CA, USA, and MGI Pharma Inc, Bloomington, MN, USA), have had a significant impact on the treatment paradigm of myelodysplastic syndromes (MDSs), previously managed mainly by supportive care and hematopoietic-stem-cell transplantation. The positive clinical experience seen in MDS to date coupled with the persistent challenges faced in the treatment of other hematologic malignancies has served as the impetus for further exploration of the therapeutic value of DNMT inhibitors beyond MDS. In that respect, the majority of data for these agents are in the setting of acute myelogenous leukemia (AML). Experience with these agents in patients with refractory anemia with excess blasts in transformation (reclassified by the World Health Organization as AML) was also reported in the clinical trials submitted to the FDA for approval of azacitidine for MDS. Some use has also been described in chronic myelogenous leukemia and acute lymphocytic leukemia. Further studies are needed to clarify the appropriate dose and the number and duration of cycles in the treatment of leukemias, and to identify ideal candidates for therapy, explore the role of DNMT inhibitors in combination with other agents, especially histone deacetylase inhibitors, delineate differences between the commercially available agents, and establish the long term safety of these agents. To this end, experience with DNMT inhibitors in hematologic malignancies other than MDS is reviewed in an effort to better understand the therapeutic potential of these agents and to define areas of future exploration in these settings. PMID- 16341240 TI - DNA methylation and gene silencing in cancer. AB - Epigenetic changes such as DNA methylation act to regulate gene expression in normal mammalian development. However, promoter hypermethylation also plays a major role in cancer through transcriptional silencing of critical growth regulators such as tumor suppressor genes. Other chromatin modifications, such as histone deacetylation and chromatin-binding proteins, affect local chromatin structure and, in concert with DNA methylation, regulate gene transcription. The DNA methylation inhibitors azacitidine and decitabine can induce functional re expression of aberrantly silenced genes in cancer, causing growth arrest and apoptosis in tumor cells. These agents, along with inhibitors of histone deacetylation, have shown clinical activity in the treatment of certain hematologic malignancies where gene hypermethylation occurs. This review examines alteration in DNA methylation in cancer, effects on gene expression, and implications for the use of hypomethylating agents in the treatment of cancer. PMID- 16341241 TI - Lipid microarrays identify key mediators of autoimmune brain inflammation. AB - Recent studies suggest that increased T-cell and autoantibody reactivity to lipids may be present in the autoimmune demyelinating disease multiple sclerosis. To perform large-scale multiplex analysis of antibody responses to lipids in multiple sclerosis, we developed microarrays composed of lipids present in the myelin sheath, including ganglioside, sulfatide, cerebroside, sphingomyelin and total brain lipid fractions. Lipid-array analysis showed lipid-specific antibodies against sulfatide, sphingomyelin and oxidized lipids in cerebrospinal fluid (CSF) derived from individuals with multiple sclerosis. Sulfatide-specific antibodies were also detected in SJL/J mice with acute experimental autoimmune encephalomyelitis (EAE). Immunization of mice with sulfatide plus myelin peptide resulted in a more severe disease course of EAE, and administration of sulfatide specific antibody exacerbated EAE. Thus, autoimmune responses to sulfatide and other lipids are present in individuals with multiple sclerosis and in EAE, and may contribute to the pathogenesis of autoimmune demyelination. PMID- 16341242 TI - Glycogen synthase kinase-3 is an in vivo regulator of hematopoietic stem cell repopulation. AB - The in vivo regulation of hematopoietic stem cell (HSC) function is poorly understood. Here, we show that hematopoietic repopulation can be augmented by administration of a glycogen synthase kinase-3 (GSK-3) inhibitor to recipient mice transplanted with mouse or human HSCs. GSK-3 inhibitor treatment improved neutrophil and megakaryocyte recovery, recipient survival and resulted in enhanced sustained long-term repopulation. The output of primitive Lin(-)c Kit(+)Sca-1(+) cells and progenitors from HSCs increased upon GSK-3 inhibitor treatment without altering secondary repopulating ability, suggesting that the HSC pool is maintained while overall hematopoietic reconstitution is increased. GSK-3 inhibitors were found to modulate gene targets of Wnt, Hedgehog and Notch pathways in cells comprising the primitive hematopoietic compartment without affecting mature cells. Our study establishes GSK-3 as a specific in vivo modulator of HSC activity, and suggests that administration of GSK-3 inhibitors may provide a clinical means to directly enhance the repopulating capacity of transplanted HSCs. PMID- 16341243 TI - Hypoxia-inducible factor determines sensitivity to inhibitors of mTOR in kidney cancer. AB - Inhibitors of the kinase mammalian target of rapamycin (mTOR) have shown sporadic activity in cancer trials, leading to confusion about the appropriate clinical setting for their use. Here we show that loss of the Von Hippel-Lindau tumor suppressor gene (VHL) sensitizes kidney cancer cells to the mTOR inhibitor CCI 779 in vitro and in mouse models. Growth arrest caused by CCI-779 correlates with a block in translation of mRNA encoding hypoxia-inducible factor (HIF1A), and is rescued by expression of a VHL-resistant HIF1A cDNA lacking the 5' untranslated region. VHL-deficient tumors show increased uptake of the positron emission tomography (PET) tracer fluorodeoxyglucose (FDG) in an mTOR-dependent manner. Our findings provide preclinical rationale for prospective, biomarker-driven clinical studies of mTOR inhibitors in kidney cancer and suggest that FDG-PET scans may have use as a pharmacodynamic marker in this setting. PMID- 16341244 TI - Comparison of the properties of the five soluble guanylyl cyclase subunits in Drosophila melanogaster. AB - The Drosophila melanogaster genome contains 5 genes that code for soluble guanylyl cyclase subunits. Two of these genes code for subunits, Gycalpha-99B and Gycbeta-100B, which form a conventional NO-sensitive guanylyl cyclase and the other three code for atypical subunits, Gyc-88E, Gyc-89Da and Gyc-89Db. The properties and distribution of Gyc-88E and Gyc-89Db have previously been described and here Gyc-89Da is described. Gyc-89Da only forms an active guanylyl cyclase when co-expressed with Gyc-88E. The three atypical subunits probably form two different heterodimers in vivo: Gyc-88E/89Da and Gyc-88E/89Db. Both of these heterodimers were slightly stimulated by NO donors and Gyc-88E/89Da showed a greater activation by Mn2+, with an increase in Vmax and a decrease in K(m), compared to Gyc-88E/89Db. Both Gyc-88E/89Da and Gyc-88E/89Db were expressed in neurons in both the peripheral and central nervous system. Although all three heterodimeric soluble guanylyl cyclases in D. melanogaster can be activated by NO and inhibited by ODQ, the atypical enzymes can be distinguished from the conventional soluble guanylyl cyclase by their sensitivity to the NO-independent activators YC-1 and BAY 41-2272, which will only activate the conventional enzyme. PMID- 16341245 TI - Interactions among three species of cereal aphids simultaneously infesting wheat. AB - Interactions among greenbug, Schizaphis graminum (Rondani), Russian wheat aphid, Diuraphis noxia (Mordvilko), and bird cherry-oat aphid Rhopalosiphum padi (L.) were examined on wheat plants (Triticum aestivum L., cultivar TAM 107). Nymphs were released on the plants as conspecific and heterospecific pairs of either first or fourth instars and evaluated for survival, developmental time, fecundity, intra-plant movement, and affinity to plant tissues. Survival from first instar to onset of reproduction averaged 90-100% across all pair combinations. Diuraphis noxia developed faster as conspecifics than in any heterospecific combination, and faster as conspecifics feeding on the same plant tissue than on different tissues. Fecundity of S. graminum was higher for conspecifics that developed on the same plant tissue than for those feeding separately. There was evidence of amensalism (one species was harmed while the other was unaffected) in that D. noxia experienced delayed development feeding in tandem with S. graminum, and reduced fecundity with both S. graminum and R. padi. Furthermore, S. graminum nymphs had reduced survival when their mothers matured on a same plant with R. padi. Both D. noxia and R. padi changed position on the plant more often when developing with S. graminum. Survival of second generation S. graminum nymphs was reduced when this species developed and reproduced in tandem with R. padi. Preferred feeding locations were S. graminum--primary leaf, D. noxia--tertiary leaf and R. padi--stem and these were not altered in any heterospecific combinations. Heterospecific aphids had no impact on fecundity or progeny survival in any species combination when fourth instars matured and reproduced on plants not previously exposed to aphid feeding, supporting the inference that systemic, aphid-induced changes in plant physiology mediated the effects observed when first instars developed and reproduced on the same plants. PMID- 16341246 TI - Role of neonicotinyl insecticides in Washington apple integrated pest management. Part I. Control of lepidopteran pests. AB - Three neonicotinyl insecticides, acetamiprid, thiacloprid and clothianidin, were evaluated for their impact on four species of lepidopteran pests of apple in Washington, the codling moth, Cydia pomonella (L.), the Pandemis leafroller, Pandemis pyrusana Kearfott, and the obliquebanded leafroller, Choristoneura rosaceana (Harris), and Lacanobia subjuncta (Grote & Robinson). None of the neonicotinyl insecticides demonstrated sufficient activity against P. pyrusana, C. rosaceana, or L. subjuncta to warrant field trials. Conversely, all had some activity against one or more stages of C. pomonella. Acetamiprid was highly toxic to larvae in laboratory bioassays, and had relatively long activity of field-aged residues (21 days). It also showed some toxicity to C. pomonella eggs (via topical exposure) and adults. Acetamiprid provided the highest level of fruit protection from C. pomonella attack in field trials conducted over five years in experimental orchards with extremely high codling moth pressure. Thiacloprid performed similarly in bioassays, but fruit protection in field trials was slightly lower than acetamiprid. Clothianidin showed moderate to high toxicity in bioassays, depending on the C. pomonella stage tested, but poor fruit protection from attack in field trials. None of the neonicotinyl insecticides were as toxic to larvae or effective in protecting fruit as the current standard organophosphate insecticide used for C. pomonella control, azinphosmethyl. However, both acetamiprid and thiacloprid should provide acceptable levels of C. pomonella control in commercial orchards where densities are much lower than in the experimental orchards used for our trials. The advantages and disadvantages of the neonicotinyl insecticides as replacements for the organophosphate insecticides and their role in a pest management system for Washington apple orchards are discussed. PMID- 16341247 TI - Oviposition deterrent and skin repellent activities of Solanum trilobatum leaf extract against the malarial vector Anopheles stephensi. AB - The leaf extract of Solanum trilobatum (Solanaceae) was tested under laboratory conditions for oviposition deterrent and skin repellent activities against the adult mosquito Anopheles stephensi. Concentrations of 0.01, 0.025, 0.05, 0.075 and 0.1% reduced egg laying by gravid females from 18 to 99% compared to ethanol treated controls. In skin repellent tests, concentrations of 0.001, 0.005, 0.01, 0.015, and 0.02 % provided 70 to 120 minutes protection against mosquito bites, whereas the ethanol control provided only 2.2 minutes of protection. Both oviposition deterrent and skin repellent activity were dose dependent. The results suggest that the leaf extract of S. trilobatum is an effective oviposition deterrent and skin repellent against An. stephensi. PMID- 16341248 TI - Role of neonicotinyl insecticides in Washington apple integrated pest management. Part II. Nontarget effects on integrated mite control. AB - The effect of neonicotinyl insecticides on integrated mite control in Washington apple was examined from 0 In a series of 20 field trials (54 treatments) designed primarily to look at efficacy against the codling moth, Cydia pomonella, nearly half of the treatments using four or more applications of acetamiprid had peak mite densities exceeding the economic threshold of 5 mites per leaf. Overall, acetamiprid treatments had 4.6-fold higher mite densities than the standard organophosphate insecticide treatment. Of the treatments with high mite populations, Panonychus ulmi, the European red mite, and Tetranychus urticae, the twospotted spider mite, were the dominant species in roughly equal numbers of cases. Only 11.1% of the thiacloprid treatments exceeded 5 mites per leaf; these experimental treatments included eight applications, whereas the current label restricts the number of applications at the rate for C. pomonella to two applications. One out of six clothianidin treatments caused a significantly higher mite density than the standard treatment; however, this material appeared to suppress predatory mites. Neonicotinyl insecticides did not eliminate predatory mites, but they inhibited their ability to respond normally to increasing prey populations. In field trials designed specifically to examine mite population densities where neonicotinyl insecticides were used, significantly higher levels of tetranychid mites occurred in one or more acetamiprid treatments (one, two or four applications) in five out of six trials. In the sixth trial (in a commercial orchard), only two acetamiprid applications were made, and mite populations were low in all treatments. While elevated mite densities were more likely to occur with four applications, in one case it occurred following a single application. The predominant tetranychid mite species (either P. ulmi or T. urticae) varied from trial to trial; however, there was no apparent bias regarding stimulation of the two species. Horticultural mineral oil was used with acetamiprid in some trials in an attempt to mitigate mite outbreaks. However, the addition of oil did not counteract the tendency of acetamiprid to increase tetranychid mite populations, and in one trial, had a negative effect on predatory mite densities. Seasonal tetranychid mite density was positively related to the total grams AI (or number of applications) of acetamiprid, thus reducing the number of applications per season should lower the probability of mite outbreaks. PMID- 16341249 TI - The Bmdsx transgene including trimmed introns is sex-specifically spliced in tissues of the silkworm, Bombyx mori. AB - Bmdsx is an orthologue of the sex-determining gene doublesex (dsx) and known to be sex-specifically expressed in various tissues of the silkworm, Bombyx mori. Its pre-mRNA is sex-specifically spliced and encodes female-specific or male specific polypeptides. The open reading frame of Bmdsx consists of 5 exons, of which exons 3 and 4 are female-specific and its pre-mRNA was known to undergo default processing to generate the female-type mRNA. Previous reports have shown that the mechanism of splicing of the doublesex gene is different in Drosophila melanogaster and Bombyx mori. However, intron 4 is so long that it is difficult to identify the intronic cis-element(s) required for male-specific splicing of Bmdsx pre-mRNA using Bmdsx minigenes whose introns are shortened in various manners. As a first step toward discovery of the cis-element, the Bmdsx mini gene, which consisted of exon 1 and 5 and internally shortened introns 2 to 4, was constructed, and transgenic silkworms expressing this construct were generated. Bmdsx pre-mRNA transcribed derived from transgene was sex-specifically spliced. This result shows that the mini gene contained the information necessary for the correct regulation of alternative splicing. PMID- 16341250 TI - The social environment affects mate guarding behavior in Japanese beetles, Popillia japonica. AB - The effect of the social environment on post-copulatory mate guarding duration in Japanese beetles, (Popillia japonica Newman), was examined in a laboratory experiment. The mate guarding durations of beetles in different sex ratios and densities were observed for 10 hours. Guarding duration was longer when females were larger, suggesting the presence of 'cryptic' male mate choice. Densities, but not sex ratio, affected the duration of guarding bouts, with males guarding for longer at higher densities. This result implies that males increase their guarding duration under conditions in which their female may be likely to be encountered by other males. The lack of a sex ratio effect on the duration of guarding bouts is consistent with other studies on this species that indicate males have difficulty distinguishing females from males. Consequently, because the sex ratio on food plants is typically male-biased, a paired male may react just to density, treating surrounding individuals as if they were mostly males. The total amount of time males spent guarding was lower at lower densities and at male-biased sex ratios; this suggests that after ceasing to guard one female, males were less able to find a subsequent mate under these conditions. PMID- 16341251 TI - Two differentially expressed ommochrome-binding protein-like genes (obp1 and obp2) in larval fat body of the European corn borer, Ostrinia nubilalis. AB - Ommochrome-binding proteins function in coloration and detoxification pathways by transporting tryptophan metabolites, and increase in hemolymph concentration prior to diapause. Two ommochrome-binding protein genes from the European corn borer Ostrinia nubilalis (Hubner) (Onobp1 and Onobp2; GenBank accession nos. AY819651 to AY819655 and AY862870) were isolated. Relatedness to OBP-encoding genes was suggested by peptide similarity, phylogenetic reconstruction, and expression data. 21 single nucleotide polymorphisms between obp1 and 23 polymorphisms between obp2 alleles were identified, and resultant genomic markers were inherited in a Mendelian fashion. RT-PCR showed fat body specific Onobp1 and Onobp2 transcription. The Onobp1 transcript was RT-PCR amplified from fat body of 5 instars, whereas Onobp2 was expressed in fat body of 4 and 5 instars, and peaked in 5 instar wandering and 1 week old diapausing larvae. Expression suggests gene duplicates are maintained by change in temporal expression. The significance of Onobp1 and 2 gene products to O. nubilalis diapause physiology requires additional investigation. PMID- 16341252 TI - Comparison of immunoreactivity to serotonin, FMRFamide and SCPb in the gut and visceral nervous system of larvae, pupae and adults of the yellow fever mosquito Aedes aegypti. AB - In all life stages, the gut of the mosquito is innervated by a small number (typically 4) of central neurons immunoreactive to serotonin (SI). The serotonergic system appears to pass through metamorphosis largely intact, despite extensive remodeling of the gut. Axons immunoreactive to antibodies raised against molluscan FMRFamide (RF-I) constitute peptidergic innervation that anatomically parallels the serotonergic system. In the larva, two clusters of 3 neurons project to the anterior regions of the gut, whereas in the pupa and adult, typically two large RF-I neurons located next to the esophagus send several processes posteriorly. In adults, these neurons branch throughout the diverticula and anterior stomach. In pupae, but not in larvae or adults, the gut RF-l system coexpresses reactivity to antibodies raised against a member of another peptide family, molluscan small cardioactive peptide b (SCP-I). SCP-I immunoreactivity is localized independently of RF-l immunoreactivity in the ganglia of all stages and in neurons that project along the gut of the adult. We did not find any colocalization of S-I and the peptide markers. Distinct populations of enteroendocrine cells populate different regions of the gut at different life stages. Changes in staining pattern suggest that these cells are replaced at metamorphosis along with the other gut cells during the extensive remodeling of the tract. Distributed in the gut epithelium are subpopulations that express either RF-I or SCP-I; a small fraction of these cells bind antibodies to both peptides. The stomachs of adult females are larger than those of males, and the numbers of SCP-I and RF-I enteroendocrine cells are proportionately greater in females. In all the life stages, the junctions between different regions of the gut are the focus of regulatory input. The larval cardiac valve possesses a ring of cells, the necklace cells, which appear to receive extensive synaptic inputs from both the serotonergic system and the peptidergic system. Another focus of control is the pyloric valve, which is encircled by axon-like processes. The immunoreactive pattern of this region differs across life stages, expressing SCP-I in larvae, S-I in pupae, and both SCP-I and RF-I in adults. PMID- 16341253 TI - Pectinmethylesterase from the rice weevil, Sitophilus oryzae: cDNA isolation and sequencing, genetic origin, and expression of the recombinant enzyme. AB - A cDNA clone encoding pectinmethylesterase of the rice weevil, Sitophilus oryzae (L.) has been isolated and sequenced. The cDNA clone was expressed in cultured insect cells and active pectinmethylesterase was purified from the culture medium, thus confirming that the cDNA encodes pectinmethylesterase. In situ hybridization indicated that the enzyme's transcript was present in the midgut. Weevils treated with tetracycline so that they lack genes of known symbiotic organisms still contained the pectinmethylesterase gene, indicating that the gene is encoded by the rice weevil genome. The rice weevil enzyme is most similar in sequence to bacterial pectinmethylesterases. Given this and the enzyme's apparently rather general absence from animal species, we suggest the possibility that this gene was transferred horizontally to an ancient weevil, possibly from a bacterial symbiont, and exists in Sitophilus species now as a result of that ancestral horizontal transfer. PMID- 16341254 TI - Functional and genetic analysis of coronavirus replicase-transcriptase proteins. AB - The coronavirus replicase-transcriptase complex is an assembly of viral and cellular proteins that mediate the synthesis of genome and subgenome-sized mRNAs in the virus-infected cell. Here, we report a genetic and functional analysis of 19 temperature-sensitive (ts) mutants of Murine hepatitis virus MHV-A59 that are unable to synthesize viral RNA when the infection is initiated and maintained at the non-permissive temperature. Both classical and biochemical complementation analysis leads us to predict that the majority of MHV-A59 ORF1a replicase gene products (non-structural proteins nsp1-nsp11) form a single complementation group (cistron1) while the replicase gene products encoded in ORF1b (non-structural proteins nsp12-nsp16) are able to function in trans and comprise at least three, and possibly five, further complementation groups (cistrons II-VI). Also, we have identified mutations in the non-structural proteins nsp 4, nsp5, nsp10, nsp12, nsp14, and nsp16 that are responsible for the ts phenotype of eight MHV-A59 mutants, which allows us to conclude that these proteins are essential for the assembly of a functional replicase-transcriptase complex. Finally, our analysis of viral RNA synthesis in ts mutant virus-infected cells allows us to discriminate three phenotypes with regard to the inability of specific mutants to synthesize viral RNA at the non-permissive temperature. Mutant LA ts6 appeared to be defective in continuing negative-strand synthesis, mutant Alb ts16 appeared to form negative strands but these were not utilized for positive-strand RNA synthesis, and mutant Alb ts22 was defective in the elongation of both positive- and negative-strand RNA. On the basis of these results, we propose a model that describes a pathway for viral RNA synthesis in MHV-A59-infected cells. Further biochemical analysis of these mutants should allow us to identify intermediates in this pathway and elucidate the precise function(s) of the viral replicase proteins involved. PMID- 16341256 TI - Genome science: a video tour of the Washington University Genome Sequencing Center for high school and undergraduate students. AB - Sequencing of the human genome has ushered in a new era of biology. The technologies developed to facilitate the sequencing of the human genome are now being applied to the sequencing of other genomes. In 2004, a partnership was formed between Washington University School of Medicine Genome Sequencing Center's Outreach Program and Washington University Department of Biology Science Outreach to create a video tour depicting the processes involved in large-scale sequencing. "Sequencing a Genome: Inside the Washington University Genome Sequencing Center" is a tour of the laboratory that follows the steps in the sequencing pipeline, interspersed with animated explanations of the scientific procedures used at the facility. Accompanying interviews with the staff illustrate different entry levels for a career in genome science. This video project serves as an example of how research and academic institutions can provide teachers and students with access and exposure to innovative technologies at the forefront of biomedical research. Initial feedback on the video from undergraduate students, high school teachers, and high school students provides suggestions for use of this video in a classroom setting to supplement present curricula. PMID- 16341257 TI - Teaching more by lecturing less. AB - We carried out an experiment to determine whether student learning gains in a large, traditionally taught, upper-division lecture course in developmental biology could be increased by partially changing to a more interactive classroom format. In two successive semesters, we presented the same course syllabus using different teaching styles: in fall 2003, the traditional lecture format; and in spring 2004, decreased lecturing and addition of student participation and cooperative problem solving during class time, including frequent in-class assessment of understanding. We used performance on pretests and posttests, and on homework problems to estimate and compare student learning gains between the two semesters. Our results indicated significantly higher learning gains and better conceptual understanding in the more interactive course. To assess reproducibility of these effects, we repeated the interactive course in spring 2005 with similar results. Our findings parallel results of similar teaching style comparisons made in other disciplines. On the basis of this evidence, we propose a general model for teaching large biology courses that incorporates interactive engagement and cooperative work in place of some lecturing, while retaining course content by demanding greater student responsibility for learning outside of class. PMID- 16341258 TI - The use of a knowledge survey as an indicator of student learning in an introductory biology course. AB - A knowledge survey (KS) is a series of content-based questions sequenced in order of presentation during a course. Students do not answer the questions; rather, they rank their confidence in their ability to answer each question. A 304 question KS was designed and implemented for a multisection, multi-instructor introductory biology course to determine whether this tool could be used to assess student learning. The KS was administered during the first 2 wk and the last 2 wk of the semester online via WebCT. Results were scored using one point for each "not confident" response (level 1), two points for each "possibly confident" response (level 2), and three points for each "confident" response (level 3). We found that scores increased significantly between the pre- and post KS, indicating that student confidence in their knowledge of the course material increased over the semester. However, the correlation between student confidence and final grades was negligible or low, and chi-square tests show that KS scores and matched exam questions were not significantly related. We conclude that under the conditions implemented in our study, the KS does not reliably measure student learning as measured by final grades or exam questions. PMID- 16341259 TI - Teaching systems biology: an active-learning approach. AB - With genomics well established in modern molecular biology, recent studies have sought to further the discipline by integrating complementary methodologies into a holistic depiction of the molecular mechanisms underpinning cell function. This genomic subdiscipline, loosely termed "systems biology," presents the biology educator with both opportunities and obstacles: The benefit of exposing students to this cutting-edge scientific methodology is manifest, yet how does one convey the breadth and advantage of systems biology while still engaging the student? Here, I describe an active-learning approach to the presentation of systems biology. In graduate classes at the University of Michigan, Ann Arbor, I divided students into small groups and asked each group to interpret a sample data set (e.g., microarray data, two-hybrid data, homology-search results) describing a hypothetical signaling pathway. Mimicking realistic experimental results, each data set revealed a portion of this pathway; however, students were only able to reconstruct the full pathway by integrating all data sets, thereby exemplifying the utility in a systems biology approach. Student response to this cooperative exercise was extremely positive. In total, this approach provides an effective introduction to systems biology appropriate for students at both the undergraduate and graduate levels. PMID- 16341260 TI - Teaching research integrity and bioethics to science undergraduates. AB - Undergraduate students in the Department of Biomedical Sciences at the University of South Alabama, Mobile, are required to take a course entitled "Issues in Biomedical Sciences," designed to increase students' awareness about bioethical questions and issues concerning research integrity. This paper describes the main features of this course and summarizes the results of a survey designed to evaluate the students' perceptions about the course. A summary of this study was presented at the 2002 Conference on Research Integrity in Potomac, MD, sponsored by the Office of Research Integrity of the National Institutes of Health. PMID- 16341261 TI - Information literacy in biology education: an example from an advanced cell biology course. AB - Information literacy skills are critically important for the undergraduate biology student. The ability to find, understand, evaluate, and use information, whether from the scientific literature or from Web resources, is essential for a good understanding of a topic and for the conduct of research. A project in which students receive information literacy instruction and then proceed to select, update, and write about a current research topic in an upper-level cell biology course is described. Students research the chosen topic using paper and electronic resources, generate a list of relevant articles, prepare abstracts based on papers read, and, finally, prepare a "state-of-the-art" paper on the topic. This approach, which extends over most of one semester, has resulted in a number of well-researched and well-written papers that incorporate some of the latest research in cell biology. The steps in this project have also led to students who are prepared to address future projects on new and complex topics. The project is part of an undergraduate course in cell biology, but parts of the assignments can be modified to fit a variety of subject areas and levels. PMID- 16341262 TI - Uterine sensitization-associated gene-1 (USAG-1), a novel BMP antagonist expressed in the kidney, accelerates tubular injury. AB - Dialysis dependency is one of the leading causes of morbidity and mortality in the world, and once end-stage renal disease develops, it cannot be reversed by currently available therapy. Although administration of large doses of bone morphogenetic protein-7 (BMP-7) has been shown to repair established renal injury and improve renal function, the pathophysiological role of endogenous BMP-7 and regulatory mechanism of its activities remain elusive. Here we show that the product of uterine sensitization-associated gene-1 (USAG1), a novel BMP antagonist abundantly expressed in the kidney, is the central negative regulator of BMP function in the kidney and that mice lacking USAG-1 (USAG1 mice) are resistant to renal injury. USAG1 mice exhibited prolonged survival and preserved renal function in acute and chronic renal injury models. Renal BMP signaling, assessed by phosphorylation of Smad proteins, was significantly enhanced in USAG1 mice with renal injury, indicating that the preservation of renal function is attributable to enhancement of endogenous BMP signaling. Furthermore, the administration of neutralizing antibody against BMP-7 abolished renoprotection in USAG1 mice, indicating that USAG-1 plays a critical role in the modulation of renoprotective action of BMP and that inhibition of USAG-1 is a promising means of development of novel treatment for renal diseases. PMID- 16341263 TI - Anti-Abeta42- and anti-Abeta40-specific mAbs attenuate amyloid deposition in an Alzheimer disease mouse model. AB - Accumulation and aggregation of amyloid beta peptide 1-42 (Abeta42) in the brain has been hypothesized as triggering a pathological cascade that causes Alzheimer disease (AD). To determine whether selective targeting of Abeta42 versus Abeta40 or total Abeta is an effective way to prevent or treat AD, we compared the effects of passive immunization with an anti-Abeta42 mAb, an anti-Abeta40 mAb, and multiple Abeta(1-16) mAbs. We established in vivo binding selectivity of the anti-Abeta42 and anti-Abeta40 mAbs using novel TgBRI-Abeta mice. We then conducted a prevention study in which the anti-Abeta mAbs were administered to young Tg2576 mice, which have no significant Abeta deposition, and therapeutic studies in which mAbs were administered to Tg2576 or CRND8 mice with modest levels of preexisting Abeta deposits. Anti-Abeta42, anti-Abeta40, and anti Abeta(1-16) mAbs attenuated plaque deposition in the prevention study. In contrast, anti-Abeta42 and anti-Abeta40 mAbs were less effective in attenuating Abeta deposition in the therapeutic studies and were not effective in clearing diffuse plaques following direct injection into the cortex. These data suggest that selective targeting of Abeta42 or Abeta40 may be an effective strategy to prevent amyloid deposition, but may have limited benefit in a therapeutic setting. PMID- 16341264 TI - Integration of flow-dependent endothelial phenotypes by Kruppel-like factor 2. AB - In the face of systemic risk factors, certain regions of the arterial vasculature remain relatively resistant to the development of atherosclerotic lesions. The biomechanically distinct environments in these arterial geometries exert a protective influence via certain key functions of the endothelial lining; however, the mechanisms underlying the coordinated regulation of specific mechano activated transcriptional programs leading to distinct endothelial functional phenotypes have remained elusive. Here, we show that the transcription factor Kruppel-like factor 2 (KLF2) is selectively induced in endothelial cells exposed to a biomechanical stimulus characteristic of atheroprotected regions of the human carotid and that this flow-mediated increase in expression occurs via a MEK5/ERK5/MEF2 signaling pathway. Overexpression and silencing of KLF2 in the context of flow, combined with findings from genome-wide analyses of gene expression, demonstrate that the induction of KLF2 results in the orchestrated regulation of endothelial transcriptional programs controlling inflammation, thrombosis/hemostasis, vascular tone, and blood vessel development. Our data also indicate that KLF2 expression globally modulates IL-1beta-mediated endothelial activation. KLF2 therefore serves as a mechano-activated transcription factor important in the integration of multiple endothelial functions associated with regions of the arterial vasculature that are relatively resistant to atherogenesis. PMID- 16341265 TI - CCR2 modulates inflammatory and metabolic effects of high-fat feeding. AB - The C-C motif chemokine receptor-2 (CCR2) regulates monocyte and macrophage recruitment and is necessary for macrophage-dependent inflammatory responses and the development of atherosclerosis. Although adipose tissue expression and circulating concentrations of CCL2 (also known as MCP1), a high-affinity ligand for CCR2, are elevated in obesity, the role of CCR2 in metabolic disorders, including insulin resistance, hepatic steatosis, and inflammation associated with obesity, has not been studied. To determine what role CCR2 plays in the development of metabolic phenotypes, we studied the effects of Ccr2 genotype on the development of obesity and its associated phenotypes. Genetic deficiency in Ccr2 reduced food intake and attenuated the development of obesity in mice fed a high-fat diet. In obese mice matched for adiposity, Ccr2 deficiency reduced macrophage content and the inflammatory profile of adipose tissue, increased adiponectin expression, ameliorated hepatic steatosis, and improved systemic glucose homeostasis and insulin sensitivity. In mice with established obesity, short-term treatment with a pharmacological antagonist of CCR2 lowered macrophage content of adipose tissue and improved insulin sensitivity without significantly altering body mass or improving hepatic steatosis. These data suggest that CCR2 influences the development of obesity and associated adipose tissue inflammation and systemic insulin resistance and plays a role in the maintenance of adipose tissue macrophages and insulin resistance once obesity and its metabolic consequences are established. PMID- 16341267 TI - Exploring the effect of xenon on biomembranes. AB - We report the initial findings of 100 ns molecular dynamics simulations of the role of cellular membranes in general anaesthesia. The effect of xenon on hydrated dipalmitoylphosphatidylcholine bilayers is described. The xenon atoms were found to prefer the interfacial and central regions of the bilayer. The presence of xenon was observed to lead to a small increase in the surface area, membrane thickness, and order of the acyl chains. PMID- 16341266 TI - Reversing the defective induction of IL-10-secreting regulatory T cells in glucocorticoid-resistant asthma patients. AB - We previously reported that human CD4+ Tregs secrete high levels of IL-10 when stimulated in the presence of dexamethasone and calcitriol (vitamin D3). We now show that following stimulation by allergen, IL-10-secreting Tregs inhibit cytokine secretion by allergen-specific Th2 cells in an IL-10-dependent manner. A proportion of patients with severe asthma fail to demonstrate clinical improvement upon glucocorticoid therapy, and their asthma is characterized as glucocorticoid resistant (SR, abbreviation derived from "steroid resistant"). Dexamethasone does not enhance secretion of IL-10 by their CD4+ T cells. Addition of vitamin D3 with dexamethasone to cultures of SR CD4+ T cells enhanced IL-10 synthesis to levels observed in cells from glucocorticoid-sensitive patients cultured with dexamethasone alone. Furthermore, pretreatment with IL-10 fully restored IL-10 synthesis in these cells in response to dexamethasone. Vitamin D3 significantly overcame the inhibition of glucocorticoid-receptor expression by dexamethasone while IL-10 upregulated glucocorticoid-receptor expression by CD4+ T cells, suggesting potential mechanisms whereby these treatments may overcome poor glucocorticoid responsiveness. We show here that administration of vitamin D3 to healthy individuals and SR asthmatic patients enhanced subsequent responsiveness to dexamethasone for induction of IL-10. This strongly suggests that vitamin D3 could potentially increase the therapeutic response to glucocorticoids in SR patients. PMID- 16341268 TI - Regulation of mitochondrial translation in yeast. AB - This review provides an overview of the current state of knowledge regarding the control of very unusual mechanism of mitochondrial gene expression and the structure of mitochondrial ribosomes, with emphasis on the potential of the yeast Saccharomyces cerevisiae as a model organism. PMID- 16341269 TI - Folding of spectrin's SH3 domain in the presence of spectrin repeats. AB - The multifunctional protein spectrin contains several different structural motifs, such as spectrin repeats and a SH3 domain. Both triple-helix spectrin repeats and the SH3 domain are believed to form independent structural entities. In alpha-spectrins the SH3 domain is localized to repeat 9, where it is positioned between helix B and helix C in the repeat unit. The presence of SH3 in repeat 9 decreases the thermal stability considerably of this repeat unit while another insert in helix C does not seem to affect the stability. Addition of one or two adjacent repeat units increases the thermal stability from ca 25 degrees C to 41 and 48 degrees C, respectively. Despite the differences in thermal stability, the folding properties of peptides comprising the SH3 domain only or together with one or more repeats are more or less the same. PMID- 16341270 TI - The synthesis, physicochemical properties and immunological activity of 5-amino-3 methylisoxazolo[5,4-d]4-pyrimidinone derivatives. AB - A series of 5-amino-3-methylisoxazole[5,4-d]4-pyrimidinone derivatives were obtained by reacting substituted 5-amino-3-methylisoxazol-4-carboxylic acid hydrazide with ethyl ortho-formate. The compounds were tested using the models of in vivo cellular and humoral immune response in mice and pokeweed mitogen-induced (PWM-induced) polyclonal antibody production in a culture of human peripheral blood mononuclear cells (PBMC). The compounds exhibited differential inhibitory activities in the described models, depending on the character and location of the substituted groups. We suggest that the compounds affect the early stages of the immune response. PMID- 16341271 TI - Modeling glycolipids: take one. AB - Molecular dynamics simulations of glycolipid bilayers consisting of 1,2-di-O palmitoyl-3-O-beta-D-glucosyl-sn-glycerol were performed using five different force field parameterizations. Comparing the results with experimental data revealed that only the all-atom model correctly reproduces both the phase behavior and the surface area per lipid. Only one of the united atom models studied reproduces the correct phase behavior. PMID- 16341272 TI - The role of mammalian DNA methyltransferases in the regulation of gene expression. AB - The term epigenetic modification denotes reversible traits of gene expression that do not include alterations to the DNA sequence. These epigenetic alterations are responsible for chromatin structure stability, genome integrity, modulation of tissue-specific gene expression, embryonic development, genomic imprinting and X-chromosome inactivation in females. Epigenetic changes include reversible DNA methylation and histone acetylation or methylation. The modification of mammalian genomic DNA includes the methylation at the 5-position of the cytosine (C) residue within cytosine-guanine dinucleotides (CpG), resulting in the formation of 5-methylcytosine (m5C). Regulatory DNA sequences in vertebrates often have little or no methylation. The methylation of mammalian genomic DNA is catalyzed by DNA methyltransferases (DNMTs), which play a special role in the initiation of chromatin remodeling and gene expression regulation. The mammalian DNMTs are DNMT1, DNMT3A and DNMT3B, which together with accessory proteins, like DNMT3L, are responsible for methylation pattern acquisition during gametogenesis, embryogenesis and somatic tissue development. Reversible epigenetic alterations lead to selective utilization of genome information through the activation or inactivation of transcription of functional genes during gametogenesis, embryogenesis and cell differentiation. Recently, several disparate isoforms of DNMT1 were identified in human somatic and female and male germ cells. Recent advances in the investigation of DNMT function in epigenetic DNA changes have formed the basis of the understanding of various disorder etiopathogeneses, and as a result, have facilitated and enabled new therapies with respect to these diseases. PMID- 16341273 TI - Isolation of a cDNA encoding the alpha-subunit of CAAX-prenyltransferases from Catharanthus roseus and the expression of the active recombinant protein farnesyltransferase. AB - Crfta/ggt_Ia (AF525030), a cDNA encoding the ?-subunit of the two types of CaaX prenyltransferase (CaaX-PTase), i.e. protein farnesyltransferase (PFT) and type I protein geranylgeranyltransferase, was cloned from Catharanthus roseus via a PCR strategy. Crfta/ggt_Ia is 1381-bp long and bears a 999-bp open reading frame encoding a protein of 332 residues (FTA) that shares 66% identity with its Lycopersicon esculentum orthologue. Southern blot analysis revealed that FTA is encoded by a single gene copy per haploid genome. Co-expression of Crfta/ggt_Ia and Crftb encoding the beta-subunit of PFT yielded purified active recombinant PFT. This enzyme is able to prenylate proteins from C. roseus, and could be used as a potent tool for prenylated protein identification. PMID- 16341274 TI - Molecular cloning and characterization of a peroxiredoxin from Phanerochaete chrysosporium. AB - Peroxiredoxins (Prxs) are a ubiquitous family of peroxidases widely distributed among prokaryotes and eukaryotes. Here, we report on the cloning and functional characterization of a cDNA designated PcPrx-1, encoding peroxiredoxin from the white-rot fungus Phanerochaete chrysosporium. The full-length PcPrx-1 cDNA (932 bp) contains an open reading frame of 200 amino acid residues with a molecular mass of 22.1 kDa. The deduced primary structure of PcPrx-1 polypeptide shows a high level of sequence identity to other recently identified 2-cys peroxiredoxins. The recombinant PcPrx-1 protein was expressed as a histidine fusion protein in Escherichia coli and purified with a Ni2+-column. The purified protein was shown to have a protective effect against plasmid DNA cleavage by reactive oxygen species. The PcPrx-1 protein displays the ability to remove H2O2 in a ferrithiocyanate system. The results of this study suggest that PcPrx-1 may play a protective role against oxidative stress in P. chrysosporium. PMID- 16341275 TI - Repeated administration of lead decreases brain 5-HT metabolism and produces memory deficits in rats. AB - Long-term exposure to low levels of lead (Pb2+) has been shown to produce learning and memory deficits in rodents and humans. These deficits are thought to be associated with altered brain monoamine neurotransmission. Increased brain 5 HT (5-hydroxytryptamine; serotonin) activity is thought to be a prerequisite for maintaining control over the cognitive information process, and is said to have a role in learning and memory. This study was designed to investigate the effects of Pb2+ administration on brain 5-HT metabolism and memory function in rats. Rats were injected daily for three weeks with Pb2+-acetate at a dose of 100 mg/kg body weight. The assessment of memory was done using the Radial arm maze (RAM) and Passive avoidance tests. The results showed spatial working memory (SWM) deficits as well as decreased brain 5-HT metabolism. Increased serotonin activity is considered to be an indication of improved cognitive performance. The results are discussed in the context of lead-induced decreases in 5-HT metabolism playing a role in the impairment of memory. PMID- 16341276 TI - The overproduction of nitric oxide associated with neutrophilic predominance is relevant to airway mycotic infections in asthmatics undergoing prolonged glucocorticoid treatment. AB - The complex relationship between the local inflammatory response and the spread of airway mycosis during prolonged glucocorticoid therapy in bronchial asthma patients remains unclear. We assessed the ability of airway leukocytes to produce nitric oxide (NO) in relation to differential inflammatory cell counts, levels of asthma severity, and coexisting airway mycotic infections. The study was carried out on leukocytes from the induced sputa (IS) of 14 patients with asthma complicated by mycotic airway infections undergoing prolonged glucocorticoid therapy (group FcA). Three groups of subjects without airway fungal infections were also studied: 18 glucocorticoid-treated asthmatics (group cA), 11 steroid free asthmatics (group A), and 13 healthy control subjects (group H). In group FcA, both the level of spontaneous production of NO and the percentages of neutrophils in the IS were significantly higher than in all the remaining groups. Additionally, a significant positive correlation was noticed between the NO levels and both the percentages of neutrophils in the IS and the symptom intensity scores. The results suggest a possible predominant role of neutrophils in the overproduction of NO related to asthma severity and coexisting fungal infections in glucocorticoid-treated patients. PMID- 16341277 TI - The cloning and characterization of Tetrahymena pyriformis translation elongation factor 1b alpha and gamma subunits. AB - The multi-subunit eukaryotic translation elongation factor 1 (eEF1) consists of two functionally distinct parts: G-protein eEF1A and guanine nucleotide exchange factor eEF1B. Here, we report on the cloning of cDNAs of both the alpha and gamma subunits of the eEF1B from the ciliated protozoan Tetrahymena pyriformis. The open reading frame of the eEF1Bgamma cDNA encodes a 399-amino acid long polypeptide with a calculated molecular mass of 45.2 kDa. The eEF1Balpha cDNA contains an open reading frame encoding a polypeptide of 228 amino acids. The calculated molecular mass of this protein is 25.2 kDa. The overall deduced amino acid sequences of eEF1Balpha and eEF1Bgamma show a considerable homology with the families of alpha and gamma proteins from other eukaryotic organisms. We demonstrated that eEF1Bgamma is an RNA-binding protein which is able to bind to different RNAs. PMID- 16341278 TI - The metabolic profiles of transgenic cucumber lines vary with different chromosomal locations of the transgene. AB - The metabolic profiles of five transgenic cucumber lines were compared taking into consideration their transgene integration sites. The plants analyzed were homozygous and contained transgenes integrated in a single locus on chromosomes I, II, III or IV. The transgenes were preferentially located in the euchromatic regions. Each of these locations possessed a specific metabolic profile. The number of altered compounds in the transgenic lines varied between 9 and 23 of the 47 metabolites identified. These alterations seem to be specific for each independent transgene integration. However, some changes are common: a decrease in the levels of phenylalanine, aspartate, ethanolamine and pipecolate, and an increase in the level of benzoic acid. The observed effects of transgene introduction are discussed in this paper. PMID- 16341279 TI - Liposomes: an overview of manufacturing techniques. AB - During the last few decades liposomes have attracted great interest as ideal models for biological membranes as well as efficient carriers for drugs, diagnostics, vaccines, nutrients and other bioactive agents. The extensive and ever increasing literature covering the field of liposomology written by researchers with diverse backgrounds is an indication of increasing interest in this field. Many techniques and methodologies have evolved for the manufacture of liposomes, on small and large scales, since their introduction to the scientific community around 40 years ago. This article intends to provide an overview of the advantages and disadvantages of liposome preparation methods in general with particular emphasis on the heating method, developed in our laboratory, as a model technique for fast production of the lipid vesicles. PMID- 16341280 TI - The increase in mitochondrial DNA copy number in the tissues of gamma-irradiated mice. AB - Changes in the number of mitochondrial DNA (mtDNA) copies in the brain and spleen tissues of gamma-irradiated (3 Gy) mice were studied by comparative analysis of the long-extension PCR products of mtDNA (15.9 kb) and a fragment of the cluster nuclear beta-globin gene (8.7 kb) amplified simultaneously in one and the same test-tube within total DNA. The analysis showed that, compared to the nuclear beta-globin gene, an increase in mtDNA copy number (polyploidization) took place in the brain and spleen cells of mice exposed to gamma-radiation. This data led to the suggestion that the major mechanism for maintenance of the mitochondrial genome, which is constantly damaged by endogenous ROS and easily affected by ionizing radiation or other exogenous factors, is the induction of synthesis of new mtDNA copies on intact or little affected mtDNA templates because the repair systems in the mitochondria function at a low level of efficiency. PMID- 16341281 TI - Multimodal effects of available drugs for ischemic stroke. AB - Recent research demonstrates the complexity and variety of pathophysiologic processes underlying cerebral ischemia. This review will focus on the multimodal effects of the various drugs currently used for stroke prevention. The most recent clinical studies of each will also be reviewed, and the implications for clinical use and future research discussed. An evolving approach for stroke prevention will include a combination of antiplatelets, increased use of statins, ACE inhibitors and possibly vitamin therapy. PMID- 16341282 TI - Atrial fibrillation. AB - The treatment of atrial fibrillation still remains challenging in everyday practice. Even after the introduction of catheter ablation, the decision making about the type of therapy became more complex. The recently published guidelines of the American Heart Association, the American College of Cardiology and the European Society of Cardiology clearly show therapeutic approaches for different types of atrial fibrillation. The fear of thromboembolism still forces the physician to restore sinus rhythm and perform a perfect anticoagulation. Based on large studies, we are able to decide for each patient individually whether oral anticoagulation or aspirin is required. Future studies in a large cohort of patients are still necessary to clarify the value of interventional therapy in conjunction with antiarrhythmic drug therapy. These will provide patients with safe therapy and a high quality of life despite presenting paroxysmal or persistent atrial fibrillation. PMID- 16341283 TI - Cardioprotective role of the VIP signaling system. AB - Vasoactive intestinal peptide (VIP) is a 28-amino acid peptide that belongs to a family of structurally related peptide hormones including pituitary adenylate cyclase-activating peptide (PACAP). These hormones are widely distributed in the nervous system, where they act as neurotransmitters. Their biological effects are mediated by specific receptors, VPAC1 and VPAC2, which have comparable affinity for VIP and PACAP, and PAC1, which binds VIP with 1,000-fold lower affinity than PACAP. Both peptides are involved in autonomic regulation of the cardiovascular system, where they exert positive inotropic and chronotropic effects, and cause coronary vasodilatation. Additionally, PACAP inhibits proliferation of cardiac fibroblasts. Several cardiovascular diseases, such as myocardial fibrosis, heart failure, cardiomyopathy and pulmonary hypertension, have been found to be associated with changes in myocardial VIP concentration or with alteration of affinity, density and physiological responsiveness of VIP/PACAP receptors. Application of the peptides or their agonists has beneficial effect in hypertension, heart failure and myocardial fibrosis. Taken together, VIP and PACAP have beneficial effects in various pathological conditions. PMID- 16341284 TI - Recent developments in cardiovascular disease in diabetes mellitus: 41st annual meeting of the European Association for the Study of Diabetes. AB - September 10-15, 2005, the European Association for the Study of Diabetes held its 41st Annual Meeting in Athens, Greece. This meeting was devoted to new developments in understanding, diagnosis and management of diabetes mellitus and related disorders. Each day's program included plenary sessions, symposia, oral presentations, poster discussions, lectures, current issues and supportive care. This article highlights recent developments in pathophysiology, new therapeutic possibilities and preventive aspects of cardiovascular diseases in diabetes mellitus. PMID- 16341285 TI - New strategies in the treatment of hypercholesterolemia. PMID- 16341286 TI - Hot topics in cardiology: a report from the 2005 meeting of the European Society of Cardiology. PMID- 16341287 TI - Drug-induced weight gain. AB - Drug-induced weight gain is a serious side effect of many commonly used drugs leading to noncompliance with therapy and to exacerbation of comorbid conditions related to obesity. Improved glycemic control achieved by insulin, insulin secretagogues or thiazolidinedione therapy is generally accompanied by weight gain. It is a problematic side effect of therapy due to the known deleterious effect of weight gain on glucose control, increased blood pressure and worsening lipid profile. Weight gain may be lessened or prevented by adherence to diet and exercise or combination therapy with metformin. Weight gain is also common in psychotropic therapy. The atypical antipsychotic drugs (clozapine, olanzepine, risperidone and quetiapine) are known to cause marked weight gain. Antidepressants such as amitriptyline, mirtazapine and some serotonin reuptake inhibitors (SSRIs) also may promote appreciable weight gain that cannot be explained solely by improvement in depressive symptoms. The same phenomenon is observed with mood stabilizers such as lithium, valproic acid and carbamazepine. Antiepileptic drugs (AEDs) that promote weight gain include valproate, carbamazepine and gabapentin. Lamotrigine is an AED that is weight-neutral, while topiramate and zonisamide may induce weight loss. PMID- 16341288 TI - Exenatide. AB - Exenatide is the first in a new class of compounds that exhibit activity similar to the naturally occurring hormone glucagon-like peptide-1 (GLP-1). Released from cells in the gut in response to food, GLP-1 binds to pancreatic beta-cell receptors to stimulate the release of insulin. Exenatide mirrors many of the effects of GLP-1, improving glycemic control through a combination of mechanisms, which include glucose-dependent stimulation of insulin secretion, suppression of glucagon secretion, slowing of gastric emptying, reduced appetite and enhanced beta-cell function. As stimulation of insulin secretion occurs only in the presence of elevated blood glucose concentrations, the risk of hypoglycemia should be greatly reduced with exenatide. In addition to positive therapeutic effects on fasting and postprandial glucose levels, exenatide treatment is associated with significant, dose-dependent reductions in glycated hemoglobin (HbA1c) from baseline and progressive reductions in body weight. Exenatide is generally well tolerated; nausea is the most commonly reported side effect, but it can be significantly reduced when a target dose of exenatide is achieved in patients with gradual dose titration. Exenatide may enable patients with type 2 diabetes to achieve glycemic control while reducing or eliminating the risk of hypoglycemia and weight gain. These would represent significant therapeutic gains. PMID- 16341289 TI - Muraglitazar: an agent for the treatment of type 2 diabetes and associated dyslipidemia. AB - Many studies indicate that postprandial metabolic abnormalities, such as hyperglycemia and dyslipidemia, which are exaggerated and prolonged in type 2 diabetes, are important risk factors for cardiovascular disease. Different pharmacotherapies have been developed to specifically target these risk factors associated with type 2 diabetes. The peroxisome proliferator-activated receptor (PPAR) agonists, which are potent insulin sensitizers, have been the focus of much research during the past decade. Since their development, PPAR agonists have emerged as an important target for the treatment of insulin resistance and dyslipidemia. The more recent development of agonists that selectively target both the alpha and gamma PPARs has provided a potential treatment of global risk in patients with the metabolic syndrome or type 2 diabetes. Muraglitazar is a non thiazolidinedione, oxybenzylglycine dual PPARalpha/gamma agonist that is in advanced clinical development for the treatment of type 2 diabetes and its associated dyslipidemia. This article summarizes the available clinical data on the efficacy and safety of muraglitazar in patients with type 2 diabetes. PMID- 16341290 TI - Zonisamide: review of its use in epilepsy therapy. AB - Zonisamide is an antiepileptic drug used as adjunctive therapy for refractory partial seizures in adults. Because of the multiple mechanisms of action, it shows a broad spectrum of anticonvulsant activity and has been effective in several types of seizures, including partial and generalized seizures, tonic clonic seizures and absence seizures in patients unresponsive to other anticonvulsants. Myoclonic epilepsy, Lennox-Gastaut syndrome and infantile spasms have also been treated effectively with zonisamide. Recent clinical studies have demonstrated additional potential for therapeutic use in neuropathic pain, bipolar disorder, migraine, obesity, eating disorders and Parkinson's disease. Despite adverse events, zonisamide is relatively safe and well tolerated in patients, and shows low discontinuation rate. It has a good pharmacokinetic profile and a low drug interaction potential. Zonisamide is considered as a drug that effectively reduces the frequency of partial seizures. PMID- 16341291 TI - Drug-induced catatonia. AB - Catatonia is a heterogeneous syndrome that varies in etiology, presentation, course and sequelae. Initially conceptualized as a subtype of schizophrenia, catatonia is now recognized to occur not only with other psychiatric conditions but also with medical conditions and drug-induced and toxic states. While drug induced catatonia is now a recognized entity, most studies club it with catatonia due to general medical conditions or organic catatonia, thus precluding any meaningful interpretation of such cases. The literature on drug-induced catatonia mostly draws from scattered case reports. This article attempts to review the available literature in this realm and integrate the information in an attempt to explore the epidemiology, etiology, mechanism and treatment of drug-induced catatonia. PMID- 16341292 TI - Anti-inflammatory properties of drugs acting on the renin-angiotensin system. AB - It is now well established that the activation of the renin-angiotensin system (RAS) is involved in the onset and progression of cardiovascular and renal diseases, and that its main effector, angiotensin II (Ang II) has major pro inflammatory activity that induces the expression of cytokines, chemokines, adhesion molecules, growth factors and reactive oxygen species. By means of these actions, Ang II induces vascular inflammation, endothelial dysfunction and fibrosis, and participates in destabilizing atherosclerotic plaque and establishing chronic kidney diseases. Blocking the RAS by inhibiting Ang II generation or blocking angiotensin receptors reduces the morbidity and mortality associated with cardiovascular and renal disease beyond the levels due to the lowering of blood pressure, and these benefits are at least partially due to the reduction/prevention of both local and systemic inflammatory processes. The aim of this review is to describe the role of the RAS (and particularly Ang II) in initiating and maintaining these processes, and to summarize experimental and clinical evidence supporting the role of drugs acting on the RAS in preventing or modulating inflammation. PMID- 16341293 TI - Urologic manifestations of genetic diseases. AB - Many genetic disorders have genitourinary manifestations in childhood. In order to care for these children, the physician should be aware of such potential manifestations. A succinct overview of the pediatric genetic disorders that have urologic manifestations is presented to assist in the evaluation of patients, counseling of parents and treatment of urologic manifestations of childhood genetic diseases. PMID- 16341294 TI - Regional variation in self-reported heart disease prevalence in Canada. AB - BACKGROUND: Cardiovascular disease is the leading cause of death in Canada. OBJECTIVE: To provide an analysis of the self-reported prevalence of heart disease and three specific cardiac conditions--myocardial infarction (MI), angina and congestive heart failure (CHF)--in subgroups of the Canadian population. METHODS: Data from the Public Use Microdata File from Statistics Canada's 2000/2001 Canadian Community Health Survey (CCHS) were used to estimate the crude self-reported prevalence of heart disease, MI, angina and CHF in Canada. The data are reported by age and sex groups, as well as by province or territory and health region. RESULTS: Based on the 2000/2001 CCHS data, it was estimated that among Canadians 12 years of age and older, 5.0% (n=1,286,000) have heart disease, 2.1% (n=537,000) have had a heart attack, 1.9% (n=483,000) have angina and 1.0% (n=264,000) have CHF. Marked variation in the prevalence of heart disease and the other specific cardiac conditions exists across age and sex groups, and across geographical regions. The prevalence of heart disease is low among those younger than 50 years; thereafter, the prevalence of heart disease increases and is more common among men than among women. By 70 years of age, at least one in four men and one in five women report having heart disease. Large differences in the burden of heart disease were observed across provinces, territories and health regions. Comparison of the highest and lowest prevalence rates among provinces and territories revealed a 1.9-fold difference for heart disease, a 2.8-fold difference for MI, a 2.3-fold difference for angina and a 3.3-fold difference for CHF. CONCLUSIONS: Large regional differences in the prevalence of heart disease and other specific cardiac conditions were observed across Canada. These data may assist health system planners to identify those regions and population subgroups most affected by heart disease, and to support the development of heart disease prevention and treatment programs. PMID- 16341295 TI - General commentary on access to cardiovascular care in Canada: universal access, but when? Treating the right patient at the right time. AB - In 2004, the Canadian Cardiovascular Society formed an Access to Care Working Group with a mandate to use the best science and information available to establish reasonable triage categories and safe wait times for common cardiovascular services and procedures through a series of commentaries. The present commentary is the first in the series and lays out issues regarding timely access to care that are common to all cardiovascular services and procedures. The commentary briefly describes the 'right' to timely access, wait lists as a health care system management tool, and the role of the physician as patient advocate and gatekeeper. It also provides advice to funders, administrators and providers who must monitor and manage wait times to improve access to cardiovascular care in Canada and restore the confidence of Canadians in their publicly funded health care system. PMID- 16341296 TI - Supervised exercise training improves aerobic capacity and muscle strength in older women with heart failure. AB - BACKGROUND: The effect that supervised or unsupervised exercise training has on aerobic capacity (peak oxygen consumption [VO2peak]), muscle strength and quality of life in older women with heart failure remains unknown. OBJECTIVE: To examine the effect of six months (three months supervised followed by three months unsupervised) of aerobic training (AT) or combined aerobic and strength training (CAST) on VO2peak, muscle strength and quality of life in older women with heart failure. METHODS: Twenty older women (mean age +/- SD, 72+/-8 years) with clinically stable heart failure were randomly assigned to AT (n=10) or CAST (n=10). Supervised AT was performed two days per week at 60% to 70% heart rate reserve, whereas unsupervised training was performed two days per week at a rate of perceived exertion of 12 to 14 on the Borg scale. The CAST group also performed one to two sets of low-to-moderate intensity strength training two days per week. RESULTS: Supervised AT or CAST resulted in an increase in VO2peak (12%; P<0.05) and leg press strength (13%; P<0.05) that returned to baseline after unsupervised training. Vertical row strength was greater (+23%; P<0.05) after supervised CAST and remained unchanged after supervised or unsupervised AT. Supervised or unsupervised exercise training was not associated with a significant change in quality of life. CONCLUSIONS: Supervised AT or CAST are effective modes of exercise to improve VO2peak and muscle strength in older women with heart failure. However, the improvements in VO2peak and muscle strength are not maintained with unsupervised exercise training. PMID- 16341297 TI - Outcome of percutaneous hybrid coronary revascularization: bare metal stents jeopardize the benefit of sirolimus-eluting stents in the real world. AB - BACKGROUND: In an effort to contain procedural costs while limiting the risk of in-stent restenosis, hybrid percutaneous revascularization (ie, stenting with at least one sirolimus-eluting stent [SES] and at least one bare metal stent [BMS] in the same patient) is felt to be a cost-effective alternative to exclusive SES use. OBJECTIVE: To describe the outcome of hybrid percutaneous revascularization for the treatment of patients with multiple coronary artery lesions. METHODS AND RESULTS: Fifty-six patients (42 men; mean age [+/- SEM] 64+/-2) underwent hybrid stenting (average of 1.2 SES/patient and 1.3 BMS/patient). SES were used to treat lesions at higher restenotic potential, including longer lesions, smaller target vessels and bifurcation lesions (mean stent length [+/- SEM] was 21.1+/-1.2 mm for SES and 16.0+/-0.6 mm for BMS; stent diameter mean [+/- SEM] was 2.9+/-0.0 mm for SES and 3.1+/-0.1 mm for BMS; bifurcation lesions were 43% for SES and 7% for BMS; all P<0.01). At nine months of clinical follow-up, no death or myocardial infarction was reported. Twenty-one patients underwent clinically driven repeat coronary angiography at a mean (+/- SEM) of 8+/-1 of months (range two to 12 months) follow-up. Target lesion revascularization procedures were recorded in six patients (11%) for nine lesions (6%). Of these lesions, seven were categorized after blinded analysis as due to in-BMS restenosis and two to in-SES restenosis (P=0.01); three patients (5.4%) underwent reangioplasty for de novo lesions. There was one case of acute in-SES thrombosis. SES showed significantly less neointimal hyperplasia (late lumen loss was 0.4+/-0.1 mm for SES and 1.3+/ 0.1 mm for BMS; loss index was 0.15+/-0.05 for SES and 0.48+/-0.05 for BMS; all P<0.001). CONCLUSIONS: The use of SES resulted in less neointimal hyperplasia even when used to treat lesions at higher risk for restenosis based on angiographic characteristics. BMS implantation significantly limits this beneficial effect, compromising the outcome of hybrid percutaneous coronary revascularization. PMID- 16341298 TI - The incidence and prognosis of Brugada electrocardiographic pattern in the Manitoba Follow-Up Study. AB - BACKGROUND: There is a paucity of long-term follow-up data on individuals with asymptomatic Brugada electrocardiographic (ECG) pattern. OBJECTIVES: To investigate the incidence and prognosis of spontaneous Brugada ECG pattern in a prospective cohort. METHODS: The Manitoba Follow-up Study is Canada's longest running study of cardiovascular disease. Since 1948, a cohort of 3983 healthy aircrew recruits has been followed with routine medical examination, including ECG. Over a 55-year follow-up period, clinical and ECG assessments were performed every three to five years, with yearly contact to monitor vital status. The mean age of the cohort at entry and the average age of the 1375 survivors in 2003 were 31 and 83 years of age, respectively. Brugada ECG pattern was defined as ST segment elevation in at least one of leads V1 to V3 with a J wave amplitude of at least 2 mm, negative T waves, generally coved ST-T configuration, in the absence of alternative explanations. Serial ECGs of 273 subjects (6.9% of the cohort) with complete right bundle branch block at any time during follow-up were reviewed. Follow-up records pertaining to clinical course were also reviewed. RESULTS: All ECGs (in total 5665) from this cohort were reviewed. Four men had intermittent Brugada ECG pattern (lifetime incidence one per 1000): three men (all 80 years of age or older) were well on last follow-up and one had died of Alzheimer's disease. None of these men had syncope or ventricular arrhythmias documented during follow-up. CONCLUSIONS: The longevity of asymptomatic individuals in this cohort was not affected by spontaneous Brugada ECG pattern. PMID- 16341299 TI - Multiple episodes of ischemic preconditioning are not associated with loss of benefit: preliminary clinical experience. AB - BACKGROUND: As a clinical analogue of ischemic preconditioning (IP), preinfarction angina (PA) shares a well-documented protective effect in the setting of acute myocardial infarction (AMI) by reducing infarct size, preserving left ventricular function and improving prognosis. In the experimental setting, multiple cycles of IP may induce the loss of this protection. OBJECTIVE: To evaluate the effect of repeated cycles of PA on clinical outcomes in patients exhibiting a first AMI. METHODS: Seventy-four consecutive patients with AMI, in whom PA was the surrogate of experimental IP, were studied prospectively. All patients had poor or no collaterals. The patients were divided into three groups: group 1 (n=32) comprised patients without PA (control subjects); groups 2 (n=24) and 3 (n=18) comprised patients reporting one to four and more than four episodes of new-onset PA, respectively (preconditioned groups). Both of the preconditioned groups were compared with the control subjects with regard to creatine kinase-MB release, corrected Q-T interval (QTc) at discharge and major in-hospital complications. RESULTS: Compared with the control subjects, groups 2 and 3 exhibited reduced creatine kinase-MB release (75+/-26 IU/L and 85+/-22 IU/L versus 172+/-13 IU/L, P=0.004 and P=0.024, respectively), lower discharge QTc values (418+/-15 ms and 422+/-19 ms versus 443+/-38 ms, P=0.004 and P=0.031, respectively), and a reduced incidence of postinfarction angina (25% and 11% versus 44%, P<0.05), arrhythmias (0% and 0% versus 22%, P<0.05) and pulmonary edema (4% and 0% versus 28%, P<0.05). CONCLUSIONS: Regardless of the number of recurrences, IP seems to be a powerful intervention to reduce infarct size, limit QTc at discharge and improve the outcome in patients with AMI. PMID- 16341300 TI - Evolution of percutaneous endovascular interventional cardiovascular specialists: convergence of cardiac surgery and interventional cardiology. PMID- 16341301 TI - Economic implications of treatment guidelines for congestive heart failure. AB - Congestive heart failure (CHF) is the most common cause of cardiovascular hospital admission. A significant proportion of the costs of CHF is due to hospitalizations. The present study evaluated the economic impact of a modest increase in the use of angiotensin-converting enzyme (ACE) inhibitors, beta blockers, spironolactone and digoxin on CHF hospitalizations. Patients with CHF were identified through the Canadian Institute for Health Information (CIHI) database. The efficacy of ACE inhibitors, beta-blockers, spironolactone and digoxin in the first year of treatment were retrieved from the Survival and Ventricular Enlargement (SAVE) trial, a meta-analysis, the Randomized Aldactone Evaluation Study (RALES) and the Digitalis Investigation Group (DIG) trial, respectively. Cost of CHF hospitalization was based on the National List of Provincial Costs. Costs of drug treatment were based on the 2002 Alberta Health and Wellness Drug Benefit list. Physician visits for drug titration were also included in the model. A total of 85,679 patients with CHF were identified with a total of 106,130 hospital discharges. A 10% increase in use of ACE inhibitors, beta-blockers, spironolactone and digoxin would incur in a total cost due to avoidable hospital admissions of 0.4 million dollars, 1.3 million dollars, 3.7 million dollars and 1.2 million dollars, respectively. Similarly, the costs of drug treatment would be 2.2 million dollars, 1.3 million dollars, 0.3 million dollars and 0.5 million dollars, respectively. An increase in the use of the above medications would save 6.6 million dollars due to avoidable hospital admissions. The total cost of drug treatment was 4.3 million dollars, giving a net savings of 2.3 million dollars in the first year. The implementation of evidence-based therapy for CHF treatment is not only clinically efficacious, but also economically attractive. PMID- 16341302 TI - Natural course of left atrial thrombi after bioprosthetic mitral valve replacement in a patient with heparin-induced thrombocytopenia. AB - Left atrial (LA) clots involving prosthetic mitral valves are unusual complications of heparin-induced thrombocytopenia (HIT), and little is known about their natural history. A 73-year-old woman underwent bioprosthetic mitral valve replacement, which was complicated by atrial fibrillation and LA thrombus due to HIT. She was treated medically and followed up for 18 months. The present case provides unique insights into the natural history of HIT-related LA thrombus involving a bioprosthetic valve. It highlights the importance of having a high index of suspicion for this condition and avoiding the use of warfarin early in the course of the condition. PMID- 16341303 TI - Abiotrophia endocarditis: case report and review of the literature. AB - Abiotrophia is a separate genus of gram-positive cocci formerly classified as nutritionally variant streptococci. These organisms cause 1% to 2% of all infective endocarditis. Unlike Enterococcus or viridans group streptococci, these organisms do not usually grow on conventional blood agar plates. Therefore, Abiotrophia endocarditis is often falsely lumped into the category of culture negative endocarditis. Adequate diagnosis and treatment of this entity requires a high index of suspicion for the organisms, even if cultures are negative. Nutritional supplementation of the growth media and/or newer 16S ribosomal RNA gene sequencing may be necessary to identify Abiotrophia as the causative organism. Specific antimicrobial therapy and continued surveillance are needed to prevent the significant morbidity and mortality associated with this infection. PMID- 16341304 TI - The illicit sale of prescribed opioid medications in Edmonton. PMID- 16341305 TI - The big picture. PMID- 16341306 TI - Evidence of a relationship between adult attachment variables and appraisals of chronic pain. AB - OBJECTIVE: Although threat and challenge appraisals of pain have been linked to both the acute and laboratory-induced pain experience, these appraisals have not yet been explored in relation to chronic pain. In addition, although attachment theory has been separately linked to the chronic pain experience and to responses to perceived threat, it has not been explored in the context of threat and challenge appraisals of chronic pain. The present paper addresses these two main goals. METHODS: A sample of 141 participants reporting noncancer pain longer than two months in duration completed a battery of assessments before commencing treatment in one of two multidisciplinary rehabilitation programs. RESULTS: Analysis of the pain appraisal inventory produced two factors (threat and challenge), replicating earlier findings. Reliability coefficients for the two extracted scales were 0.87 and 0.86, respectively. Links between the two pain appraisal scales and adult attachment variables proved to be significant. In particular, comfort with closeness was strongly related to challenge appraisal, while anxiety over relationships was strongly associated with threat appraisal. As expected, both appraisals and attachment variables were also related to catastrophizing and emotional distress. CONCLUSIONS: Results support use of the pain appraisal inventory as an evaluation tool for those with chronic pain. Findings further suggest a substantial association between adult attachment and pain appraisal, supporting the need for more in-depth analysis and consideration of implications for treatment. PMID- 16341307 TI - Effects of flotation-restricted environmental stimulation technique on stress related muscle pain: what makes the difference in therapy--attention-placebo or the relaxation response? AB - INTRODUCTION: The purpose of the present study was to examine the potential effects of attention-placebo on flotation tank therapy. Flotation-restricted environmental stimulation technique is a method whereby an individual lies in a floating tank and all stimuli are reduced to a minimum. METHODS: Thirty-two patients were diagnosed as having stress-related muscular pain. In addition, 16 of the participants had received the diagnosis of burnout depression. The patients were treated with flotation-restricted environmental stimulation technique for six weeks. One-half of the patients were also given special attention for 12 weeks (high attention), while the remainder received attention for only six weeks (normal attention). RESULTS: The participants exhibited lowered blood pressure, reduced pain, anxiety, depression, stress and negative affectivity, as well as increased optimism, energy and positive affectivity. The results were largely unaffected by the degree of attention-placebo or diagnosis. CONCLUSION: It was concluded that flotation therapy is an effective, noninvasive method for treating stress-related pain, and that the method is not more affected by placebo than by other methods currently used in pain treatment. The treatment of both burnout depression and pain related to muscle tension constitutes a major challenge for the patient as well as the care provider, an area in which great gains can be made if the treatment is effective. Flotation therapy may constitute an integral part of such treatment. PMID- 16341308 TI - Fear of pain and fear of falling among younger and older adults with musculoskeletal pain conditions. AB - BACKGROUND: The fear-avoidance model of pain accounts for the debilitating consequences of strong fear as a component of reactions to painful conditions (eg, movements become anxiety-provoking because they cause pain). The model, which is supported in the literature, posits that fear of pain interferes with recovery because it leads to avoidance of beneficial activity. Despite the high prevalence of pain among the elderly, investigations of fear of pain in this population are scarce. A related construct, fear of falling, has been studied among elderly (but not younger) adults as an age-specific concern. OBJECTIVES: To examine the relationship between fear of pain and fear of falling. Specifically, it is hypothesized that fear of pain and fear of falling are distinct constructs. Moreover, the authors investigated whether fear of falling becomes more relevant with increasing age in a sample of seniors and younger adults with musculoskeletal conditions. METHODS: A convenience sample of younger and older adult physiotherapy outpatients (n=226) receiving treatment for musculoskeletal conditions were recruited for the study. Fear of pain and fear of falling were assessed using self-report measures. RESULTS: There were no age differences with respect to fear of pain and fear of falling. Regression analyses showed that fear of pain measures contribute substantial unique variance to the prediction of each other; however, they only make minor unique contributions to the prediction of fear of falling. Similar results were obtained through confirmatory analyses using structural modelling techniques. CONCLUSIONS: The analyses supported the distinctiveness of fear of pain and fear of falling. Moreover, among physiotherapy outpatients with musculoskeletal pain conditions, fear of falling and fear of pain are distinct constructs that appear to represent the concerns of both seniors and younger pain patients. PMID- 16341309 TI - Styles of pain coping predict cardiovascular function following a cold pressor test. AB - BACKGROUND: While many studies have examined the impact of pain-related coping in adjustment to chronic pain, relatively few have evaluated the physiological consequences of differences in pain-coping styles. OBJECTIVE: The association between strategies for coping with pain and cardiovascular functioning following exposure to painfully cold water was evaluated. METHODS: Impedance cardiography was used to assess cardiovascular functioning in 50 healthy subjects before and after a cold pressor test. RESULTS: The self-reported use of active pain-coping strategies was associated with reductions in mean arterial pressure and cardiac contractility from pre- to post-cold pressor. In contrast, higher levels of pain catastrophizing were related to increases in cardiac contractility following acute cold pain. Importantly, coping variables predicted cardiovascular responses, whereas characteristics of the noxious stimulus (pain tolerance time and pain ratings) did not. CONCLUSIONS: These findings highlight the importance of coping styles in shaping physiological responses to pain and suggest that interventions targeting increases in adaptive coping and decreases in pain catastrophizing may reduce pain's adverse impact. PMID- 16341310 TI - The role of gemcitabine in the treatment of cholangiocarcinoma and gallbladder cancer: a systematic review. AB - BACKGROUND: Cholangiocarcinoma and gallbladder cancer are difficult to treat curatively. The treatment of choice is surgery, dependent on detection at a resectable stage. No chemotherapy or radiotherapy options have shown substantial activity. Gemcitabine has demonstrated response in similar cancers. Considering the lack of treatment options for cholangiocarcinoma and gallbladder cancer, a systematic review of the evidence on gemcitabine use for these indications was performed. OBJECTIVE: To perform a systematic review to evaluate the role of gemcitabine in the treatment of cholangiocarcinoma and gallbladder cancer. METHODS: The MEDLINE database was searched (1996 to March 2005) using the medical subject headings 'gemcitabine' and 'gallbladder neoplasms' with results limited to English only. Proceedings from the 1998 to 2004 meetings of the American Society of Clinical Oncology, including the 2004 Gastrointestinal Cancers Symposium, were searched for relevant abstracts. The Canadian Medical Association infobase and the National Guidelines Clearinghouse were also searched for practice guideline reports. Reports were selected and reviewed by two reviewers, and the reference lists from those were searched for additional trials. RESULTS: A total of 13 single-arm phase II trial reports were obtained. CONCLUSIONS: In appropriate patients with gallbladder cancer or cholangiocarcinoma, surgery offers the best chance for survival and should remain the first treatment of choice. For patients not considered candidates for surgery, but willing and able to tolerate chemotherapy alone or in combination with a fluoropyrimidine (such as 5-fluorouracil or capecitabine), gemcitabine appears to be a reasonable alternative to best supportive care, although this conclusion has not been confirmed with a randomized controlled trial. PMID- 16341311 TI - The anti-Saccharomyces cerevisiae antibody assay in a province-wide practice: accurate in identifying cases of Crohn's disease and predicting inflammatory disease. AB - OBJECTIVE: To determine the utility of the anti-Saccharomyces cerevisiae antibody (ASCA) ELISA test developed in Manitoba in 2001 in a population-wide sample referred from physicians across Manitoba in their investigation of patients with gastrointestinal symptoms. METHODS: Patients whose serum was referred for ASCA testing in 2001 and 2002 were eligible for the present study. ELISA was performed by a technologist, blind to patient diagnoses. A single investigator contacted physicians to facilitate chart review. Data collected included demographics, final diagnoses and tests used to substantiate the final diagnosis. RESULTS: Of 482 subjects identified, 410 charts were available for review and 29 of those were unavailable for follow-up or had incomplete charts. The present study population included Crohn's disease (CD, n=114), ulcerative colitis (n=74), indeterminate colitis (n=31), celiac disease (n=9), irritable bowel syndrome (n=75), other diagnoses (n=33) and no disease (n=45). ASCA had a sensitivity of 37% (95% CI 27.8 to 46.8) and specificity of 97% (95% CI 93.8 to 98.6) for diagnosing CD and an odds ratio for a diagnosis of CD of 18.4 (95% CI 8.2 to 41.3). The 47 ASCA-positive patients included the following diagnoses: CD=39, ulcerative colitis=3, indeterminate colitis=1, celiac disease=3 and no disease=1. The likelihood of having an inflammatory disease if ASCA is positive was nearly 40-fold. CONCLUSION: A positive ASCA test using this assay nearly clinches a diagnosis of some form of inflammatory intestinal disease, which is highly likely to be CD. In symptomatic patients, a positive ASCA test should encourage the clinician to pursue further investigations. PMID- 16341312 TI - Intrahepatic cholangiocarcinoma presenting as the Budd-Chiari syndrome: a case report and literature review. AB - Intrahepatic cholangiocarcinoma, an increasingly recognized primary tumour of the liver, is associated with a very poor prognosis. A patient with this tumour who presented with Budd-Chiari syndrome (the first to the authors' knowledge in Western literature and only the third patient overall) secondary to extensive thrombosis in his inferior vena cava extending from the right atrium down to his iliac vessels is described. Neither curative nor palliative intervention was deemed to be an option in this patient, who deteriorated rapidly while on anti coagulants. Postmortem examination confirmed the radiological findings, and histological analysis revealed characteristic appearances of this tumour within the biliary tree and invasion into the inferior vena cava. Furthermore, biliary dysplasia, which can be a precursor to this cancer, was also noted within some of the bile ducts. PMID- 16341313 TI - Acute coinfection with hepatitis B and hepatitis C viruses. AB - Acute coinfection of hepatitis B virus (HBV) and hepatitis C virus (HCV) is rare. The few previously reported cases all describe acute HBV followed by acute HCV, leading to HBV clearance but chronic HCV. This is the first reported case of acute concurrent infection and spontaneous clearance of both HBV and HCV. PMID- 16341314 TI - Well differentiated intrahepatic cholangiocarcinoma in the setting of biliary papillomatosis: a case report and review of the literature. AB - A 64-year-old man presented with long-standing, vague, epigastric abdominal pain. History, physical examination and laboratory studies were noncontributory. However, serial computed tomography scans revealed a rapidly progressive mass in segment 2 of the liver. Surprisingly, surgical pathology revealed a well differentiated intrahepatic cholangiocarcinoma associated with biliary papillomatosis (BP). BP is a rare, benign and potentially fatal disease of the intra- and extrahepatic bile ducts. It is typified by numerous multicentric papillary fronds arising from biliary columnar epithelium. Most patients present with symptoms of jaundice and cholangitis. Although a benign disease, a review of the literature demonstrated that BP often recurs after surgical resection, carries a poor prognosis and has a moderately high malignant transformation rate. Treatment options for BP include surgical resection, transplant, ablation, stenting and/or bypass. PMID- 16341315 TI - Jejunogastric intussusception: an unusual cause of hematemesis. AB - Jejunogastric intussusception is an uncommon but potentially life-threatening complication of a previous gastrojejunal anastomosis. Although jejunogastric intussusception was first described in 1914, fewer than 200 cases have been reported in the English literature thus far. Awareness of this rare complication would help in early diagnosis and appropriate management. Described here is a case report of a patient who presented with hematemesis due to an acute jejunogastric intussusception associated with gangrene of the intussuscepted jejunum. PMID- 16341316 TI - Disclosure please! PMID- 16341317 TI - Medicine and chiropractic -- towards a better relationship. PMID- 16341319 TI - IMA mercy missions. PMID- 16341318 TI - National Injury Mortality Surveillance System -- a call for city-level injury prevention. PMID- 16341321 TI - [C Louis Leipoldt view on sexuality]. PMID- 16341320 TI - Status epilepticus -- a plea for phenobarbitone. PMID- 16341322 TI - Are health professionals aware of women's fear of rape? PMID- 16341323 TI - South Africans do sex... differently. PMID- 16341324 TI - Rest 'Boks of face World Cup defeat -- Noakes. PMID- 16341325 TI - Alleviating suffering -- one patient at a time. PMID- 16341326 TI - Blood service adjusts after 'racist' claims. PMID- 16341327 TI - The Draft Charter of the Private and Public Health Sectors of the Republic of South Africa: health for all, or profits for few? PMID- 16341328 TI - Swallowing a thermometer bulb -- is this clinical mishap dangerous? PMID- 16341329 TI - Did Adolf Hitler have syphilis? AB - The evidence that Adolf Hitler might have suffered from incapacitating syphilis is reviewed. Rumors that he acquired syphilis from a prostitute at the age of 20 years, with possible re-infection during World War I, can no longer be verified. Evidence is that he was sexually rather inactive throughout his life. Suggestions that Hitler's cardiac lesion and complaints such as transitory blindness, tremor of his left arm and leg, recurring abdominal pain and a skin lesion of the leg were of syphilitic aetiology cannot be supported. Hitler's progressive mental and physical deterioration after 1942, his growing paranoia, fits of rage, grandiosity and symptoms of possible dementia would fit in neurosyphilis. There are, however, also other explanations for his terminal syndrome, and evidence that repeated clinical examinations did not show the characteristic signs of dementia paralytica or tabes dorsalis, swings the balance of probability away from tertiary syphilis. PMID- 16341330 TI - Mercury and thermometers. PMID- 16341331 TI - Follow-up of a suspected excess of brain tumours among Namibian children. PMID- 16341332 TI - Sexually transmitted infections -- factors associated with quality of care among private general practitioners. AB - OBJECTIVES: To study the factors associated with quality of sexually transmitted infection (STI) care among private general practitioners in Gauteng. METHODS: We analysed 1 194 records of patients attending 26 randomly selected GP practices in the first 3 months of 2000 and 2002, for 3 STI syndromes, namely urethral discharge, pelvic inflammatory disease and genital ulcers. We assessed adherence to nationally accepted STI treatment guidelines and analysed the influence of patient and practice-level variables on effectiveness of STI drug regimens and trends over time. RESULTS: After controlling for syndrome mix, district and time period, appropriate drug treatment for STIs was significantly associated with the client having medical aid (p < 0.001), recent graduation as a medical practitioner (p < 0.001) and male GP gender (p = 0.007). Between 2000 and 2002, STI care improved for clients with medical aids but for not cash clients. CONCLUSIONS: There was variation in the quality of prescribing for STIs among GPs and positive trends in this prescribing. There is a need for interventions that address the financial incentives that may hamper quality of STI care for cash clients. PMID- 16341333 TI - A view of paediatric outcomes research. PMID- 16341334 TI - Audit of the outcome of pregnancy in diabetic women in Soweto, South Africa, 1992 - 2002. AB - OBJECTIVE: To audit the outcome of pregnancy among diabetic black women in Soweto following the introduction of a specialised service. DESIGN AND METHODS: An analysis of patients' records was conducted for the period 1992 - 2002. Maternal characteristics, aspects of treatment and its complications, mode of delivery and perinatal outcome are reported. The incidence of postpartum glucose intolerance in women with gestational diabetes was assessed. Major outcomes from this study are compared with those of an earlier study conducted from 1983 to 1992. RESULTS: A total of 733 diabetic pregnancies was managed over 11 years: 348 women had gestational diabetes, 172 had type 1 diabetes and 213 had type 2 diabetes. Insulin was used in all but 23 women. Mean (+/- standard deviation (SD)) capillary blood glucose levels varied between 5.7 (+/- 0.72) and 6.2 (+/- 0.95) mmol/l in the 3 groups. Severe maternal hypoglycaemia was most common in the type 1 diabetes group (14%). Mean (+/- SD) neonatal weight varied between 2 922.3 g #x0028;+/- 753) and 3 073.3 g (+/- 727). Caesarean section rates exceeded 60% in all 3 groups. Overall, 13.1% of women had hypertension, 31.3% of whom had proteinuric hypertension. There were no cases of progressive pre-eclampsia. The combined perinatal mortality (PMN) rate was 3.7%, compared with 15.6% in the 'control' group. Major congenital malformations occurred in 11 pregnancies (1.5%). There were 2 maternal deaths. Postpartum impaired glucose tolerance/diabetes occurred in 25.9% of women with gestational diabetes. Combining the 2 studies (1983 - 1992 and 1992 - 2002), 1 087 pregnancies were managed over 20 years using a similar protocol. There were no significant differences in major outcomes between the 2 studies. The overall PMN of 4.5% is significantly lower than that of the combined 'control' group (19.9%). Postpartum glucose intolerance was documented in 28.5% of women. CONCLUSIONS: The institution of a specialised service for pregnant diabetic women from Soweto for a sustained period of 20 years proved feasible and was of significant benefit. The interventions, centred around a diabetes nurse educator, were relatively simple and easy to implement. Late presentation/referral of patients remains a problem. Persistence of glucose intolerance postpartum in women with gestational diabetes was common and provides an opportunity for preventive measures to be introduced. PMID- 16341335 TI - A radiological study of the rheumatoid hand in black South Africans. AB - OBJECTIVE: To determine wrist and hand involvement in black South African patients with rheumatoid arthritis. METHODS: Larsen scoring of the wrist and hand was done in 75 patients. The mean finger score was 9.67 (range 0 - 100) on the left hand and 10.3 (range 0 - 100) on the right. Scores for the wrists were 2.5 (range 0 - 5) for the left and 2.7 (range 0 - 5) for the right. CONCLUSION: Finger and thumb involvement were considerably less in the South African black population than in other series consisting mainly of white patients. PMID- 16341336 TI - Barriers to initiating insulin therapy in patients with type 2 diabetes mellitus in public-sector primary health care centres in Cape Town. AB - BACKGROUND: The majority of patients with type 2 diabetes mellitus in Cape Town who attend primary care community health centres (CHCs) have unsatisfactory glycaemic control. Insulin is rarely prescribed despite its being indicated for type 2 diabetic patients with inadequate metabolic control on maximum oral glucose-lowering agent (OGLA) therapy. OBJECTIVE: The study examined barriers to initiating insulin therapy in poorly controlled type 2 diabetes patients on maximum OGLAs in CHCs in the Cape Town metropolis. METHODS: Five focus group discussions and 10 in-depth semistructured individual interviews were conducted with 46 medical officers working at the CHCs. The discussions and interviews were transcribed and common themes were identified and categorised. RESULTS: Doctor, patient, and system barriers to initiating insulin therapy were identified. Doctors' barriers include lack of knowledge, lack of experience with and use of guidelines related to insulin therapy, language barriers between doctor and patients, and fear of hypoglycaemia. Patient barriers were mistaken beliefs about insulin, non-compliance, lack of understanding of diabetes, use of traditional herbs, fear of injections, and poor socioeconomic conditions. System barriers were inadequate time, lack of continuity of care and financial constraints. CONCLUSION: Suggestions for overcoming barriers include further education of doctors on insulin initiation and the use of standardised guidelines. In addition, a patient-centred approach with better communication between doctors and patients, which may be achieved by reorganising aspects of the health system, may improve patient knowledge, address mistaken beliefs, improve compliance and help overcome barriers. Further research is needed to investigate these recommendations and assess patients' and nurses' perceptions on initiating insulin therapy. PMID- 16341337 TI - Community-acquired methicillin-resistant Staphylococcus aureus in Taiwan. AB - Staphylococcus aureus is a major cause of infections in both hospitals and communities, and is exhibiting increasing resistance to methicillin (methicillin resistant S. aureus, MRSA) and related beta-lactams. MRSA is usually considered a nosocomial pathogen, but increasingly it is acquired in the community. In Taiwan, MRSA was colonized in a substantial proportion of healthy children and accounted for 25% to 75% of childhood community-acquired (CA) S. aureus infections. From the preliminary data, the isolates of sequence type (ST) 59 by multilocus sequence typing method appeared to be the major clone of CA-MRSA in northern Taiwan. Compared with those reported from the US and other countries, CA-MRSA isolates in Taiwan did not always harbor type IV staphylococcal cassette chromosome (SCCmec) and were resistant to multiple non-beta-lactam antibiotics, including clindamycin and macrolides. Molecular evidence suggested transmission of the community strain of MRSA into the hospital setting, and that the community strain had became a health care-associated pathogen. The treatment of putative CA S. aureus infection should be stratified according to the severity and the disease entity. PMID- 16341338 TI - Comparison of multiple genital tract infections with Chlamydia trachomatis in different strains of female mice. AB - We have previously shown that female outbred CF-1 mice are susceptible to prolonged genital tract infection with the oculogenital serovars (D-K) of Chlamydia trachomatis, and that partial homotypic and heterotypic protection against reinfection is induced. To understand the possible role of inherent T helper 1 (Th1)/Th2 polarity bias on both the course of infection and the level of acquired immunity induced by infection, 2 immunologically different and well characterized inbred strains of mice, BALB/c and C57BL/6, were studied in this model. Groups of mice were inoculated intravaginally with C. trachomatis serovar D (Ct D) and monitored by culture to determine the duration of initial infection. Two months later, mice were reinfected, and monitored along with age- and condition-matched control groups. Plasma and vaginal secretions were collected for serologic analysis and specific delayed-type hypersensitivity was assessed by footpad swelling. Initial infection in C57BL/6 mice was comparable in duration to outbred CF-1 mice (median duration 42 versus 43.5 days), while BALB/c mice had a shorter median duration of initial infection (12 days). All strains had significantly shorter durations of infection following reinfection. BALB/c mice shed 4-10 times more inclusion-forming units (IFU) than both C57BL/6 and CF-1 mice on sample days during the first week of infection and all strains shed less IFU during reinfection. C57BL/6 and BALB/c mice had significantly lower anti-Ct D immunoglobulin G titers in both plasma and vaginal secretions than CF-1 mice following resolution of infection; the frequency of immunoglobulin A seropositive vaginal secretions was less in both inbred strains, being significantly less in the case of C57BL/6 mice. Qualitative analysis of the antigen specificity and isotype composition revealed differences among the mouse strains. All 3 strains had detectable levels of specific footpad swelling on day 14 of infection, whereas only BALB/c mice showed a significant response at 70 days post-infection. Significant differences between 2 strains of mice that differ in Th1/Th2 polarity bias were observed in: 1) the duration of infection; 2) the level of bacterial shedding during infection; and 3) the quantitative and qualitative cellular and humoral responses made in response to female genital tract infection with a human oculogenital isolate of C. trachomatis. In addition, a similar and significant level of partial acquired immunity to reinfection was observed in both strains, suggesting that inherent Th1/Th2 polarity bias present upon initial infection does not prevent the development of a protective immune response within the genital tract during infection with an oculogenital isolate of C. trachomatis. PMID- 16341339 TI - Effect of static electric field treatment on multiple antibiotic-resistant pathogenic strains of Escherichia coli and Staphylococcus aureus. AB - This study evaluated the effect of a (4.5 kV/cm, 50 Hz) static electric field (SEF) on pathogenic strains of Escherichia coli and Staphylococcus aureus with multiple antibiotic resistance. The bacteria were grown overnight at 37 degrees C in a nutrient broth medium, then inoculated in 5 mL fresh nutrient broth medium and incubated for 2 h at 25 degrees C with continuous shaking at 190 rpm. 10 x colony-forming units/mL of these bacteria were subjected to a 4.5 kV/cm, 50 Hz, SEF for various time periods. The effects of 5 different SEF exposure times (30, 60, 90, 120 and 150 min) on the bacteria were evaluated by the plate count agar method. The growth percentages of SEF treatment groups were significantly less than that of the control group. Inactivation significantly increased with the duration of SEF exposure. The results indicate that growth inhibition by SEF in the Gram-negative bacteria, E. coli, was greater than that in the Gram-positive bacteria, S. aureus. This study has demonstrated the antimicrobial effects of SEF treatment on 2 important pathogens, suggesting its potential for application as a method for controlling microbial population growth within in a variety of environments. PMID- 16341340 TI - The receptor of an oyster juice-borne coliphage OJ367 in the outer membrane of Salmonella derby. AB - The objective of this study was to identify the receptor of OJ367, an oyster juice-borne bacteriophage, in Salmonella derby ATCC 6960. The crude receptor outer membrane (OM) fraction was prepared and examined from the total cell envelope (TCE) by differential extraction with N-(2-hydroxyethyl)piperazine-N'-2 ethanesulfonic acid (HEPES)-MgCl2 and then with Triton-HEPES-ethylenediamine tetra-acetic acid buffers. The OM proteins (Omps) were isolated by diethylaminoethyl column chromatography to screen for receptor activity. A 45-kDa protein belonging to a minor Omp species, with phage neutralization ability, was eluted in a homogeneous form. It was a non-peptidoglycan-associated protein which was digestible by trypsin. Lipopolysaccharide had no influence on its receptor activity when coexistent in the diethylaminoethyl column fractions. An S. derby mutant resistant to lysis by phage OJ367 was isolated. The mutant not only showed decreased receptor activity in vitro when its TCE was tested but had an altered Omp profile. This implied that the 45-kDa Omp is involved as a receptor in coliphage binding; however, this role is affected by the expression of other Omps. PMID- 16341341 TI - Mutagenicity study of butachlor and its metabolites using Salmonella typhimurium. AB - Butachlor is the most commonly used herbicide in Taiwan and many other countries. It has been reported to be an indirect mutagen and carcinogen in various in vitro assay systems. Previous investigation has also demonstrated that butachlor stimulates cell proliferation, transforms normal embryonic cells, and induces stomach tumors in Spraque-Dawley rats. However, the mechanism of butachlor carcinogenicity is still not clear. In order to clarify the toxicologic and carcinogenic properties of butachlor, we proposed a metabolic pathway, and synthesized the authentic metabolites by chemical methods. In addition, we tested the mutagenicity of butachlor and these metabolites on Salmonella typhimurium. The results indicate that butachlor might manifest its carcinogenicity via the mutagenicity of its metabolic products. Although the molecular mechanism of butachlor-induced cellular toxicity is still not clear, it is likely that the cellular transformation ability of butachlor is partly associated with its mutagenicity. PMID- 16341342 TI - Clinical features and factors of unfavorable outcomes for non-polio enterovirus infection of the central nervous system in northern Taiwan, 1994-2003. AB - This study investigated the clinical manifestations and outcomes of central nervous system (CNS) infection by enteroviruses. Cases with CNS involvement among all enterovirus-culture-positive cases from January 1995 to June 2003 were retrospectively reviewed. Among 1028 enterovirus-culture-positive cases, there were 333 cases involving the CNS. Of these, the ratio of male to female subjects was 1.78, and the mean (+/- standard deviation) age was 6.83 +/- 5.9 years; 21 were premature neonates, and 10 failed to thrive. Disease entities included 282 cases of aseptic meningitis (84.7%), 44 cases of encephalitis (13.2%), and 7 cases of encephalomyelitis/polio-like syndrome (2.1%). Of these cases, 97.9% (326/333) had fever with peak body temperature at 38.9 degrees C, 85% had headache and vomiting, 70% had meningeal signs, 64% had neck stiffness, 16.6% (55/333) had change of consciousness, 5.4% (18/333) had seizures and 5.2% (17/333) had myoclonic jerks. Mannitol was administered in 77.2% of patients (257/333), along with intravenous immunoglobulin in 6.6% (22/333). Twelve cases received ventilator support. One patient died of hand-foot-and-mouth disease, encephalitis plus cardiopulmonary failure, and 2 premature neonates died of hepatic failure, disseminated intravascular coagulation, sepsis-like syndrome and myocarditis. Eighteen had neurologic sequelae, including 7 with limb weakness, 5 with epilepsy, 2 with sixth cranial nerve palsy, 3 with cerebral palsy, 4 with psychomotor retardation, 2 with spasticity, and 1 with hearing loss. Factors associated with unfavorable outcomes (death or sequelae) included younger age (p=0.0003), higher peak white blood cell count (WBC) [p=0.0009] and skin rash (p=0.005). Younger age and higher peak WBC were poor prognostic factors of severe enterovirus CNS infection. Death was related to neonatal enterovirus infection and enterovirus 71 infection in young children. PMID- 16341343 TI - In vitro antimicrobial effect of cefazolin and cefotaxime combined with minocycline against Vibrio cholerae non-O1 non-O139. AB - The most common clinical manifestation of Vibrio cholerae non-O1 non-O139 is gastroenteritis. This vibrion may also cause bacteremia, soft tissue infection, and other extraintestinal invasive disease, especially in immunocompromised patients. This study evaluated the current status of antimicrobial resistance in clinical isolates of V. cholerae non-O1 non-O139 in Taiwan as part of the SMART (Surveillance from Multicenter Antimicrobial Resistance in Taiwan) program. Minimal inhibitory concentrations (MICs) of 9 antimicrobial agents were determined by the agar dilution method. All of the isolates were susceptible to minocycline (MIC at which 90% of the isolates were inhibited [MIC(90)], 0.12 microg/mL), cefotaxime (MIC(90), 0.06 microg/mL), lomefloxacin (MIC(90), 0.12 microg/mL), levofloxacin (MIC(90), 0.03 microg/mL), ciprofloxacin (MIC(90), 0.03 microg/mL), moxifloxacin (MIC(90), 0.06 microg/mL), sparfloxacin (MIC(90), 0.06 microg/mL), gatifloxacin (MIC(90), 0.03 microg/mL), and cefazolin (MIC(90), 8 microg/mL). We conducted time-kill studies to evaluate the inhibitory activities of either cefazolin or minocycline alone or in combination against V. cholerae non-O1 non-O139 (Vc2). We also evaluated the inhibitory activity of cefazolin or cefotaxime combined with minocycline. The individual MICs of cefazolin, cefotaxime, and minocycline were 4 microg/mL, 0.0075 microg/mL, and 0.12 microg/mL, respectively, when approximately 5 x 105 colony-forming units/mL of V. cholerae non-O1 non-O139 was incubated. Bacterial growth was inhibited initially but resumed later when cefazolin, cefotaxime, or minocycline was used alone. When cefazoline or cefotaxime was combined with minocycline, V. cholerae non-O1 non O139 was inhibited over 48 h and no regrowth was noted. We conclude that the combination of cefazolin or cefotaxime with minocycline has a synergistic inhibitory effect on V. cholerae non-O1 non-O139 in vitro. PMID- 16341344 TI - Microbiology and factors affecting mortality in necrotizing fasciitis. AB - Necrotizing fasciitis is a life-threatening soft-tissue infection primarily involving the superficial fascia. This study investigated the microbiologic characteristics and determinants of mortality of this disease. The medical records of 87 consecutive patients with a diagnosis of necrotizing fasciitis from 1999 to 2004 were retrospectively reviewed. A single pathogen was identified as the infectious agent in 59 patients (67.8%), multiple pathogens were identified in 17 patients (19.6%), and no organism was identified in 11 patients (12.6%). Klebsiella pneumoniae, identified in 17 patients, was the most commonly isolated species. The most common comorbidity was diabetes mellitus (41 patients; 53.2%). Multivariate logistic regression analysis showed that more than 1 comorbidity, thrombocytopenia, anemia, more than 24 h delay from onset of symptoms to surgery and age greater than 60 were independently associated with mortality. This study found that K. pneumoniae was the most common cause of necrotizing fasciitis. Early operative debridement was independently associated with lower mortality. PMID- 16341345 TI - Community-acquired anaerobic bacteremia in adults: one-year experience in a medical center. AB - A prospective observational study was conducted to evaluate the clinical characteristics and outcome of community-acquired anaerobic bacteremia. From June 1 2001 through May 31 2002, 52 patients with community-acquired anaerobic bacteremia were enrolled at the emergency department in a teaching hospital. There were 19 patients (34%) with polymicrobial bacteremia and Escherichia coli was the most common copathogen (n = 6). Of 62 anaerobic isolates, species of the Bacteroides fragilis group were the most common isolates (n = 28, 45%), followed by Clostridium spp. (n = 11, 18%). Among the 52 patients enrolled, up to 27% had underlying malignancy and the gastrointestinal tract accounted for 48% of the sources of infection. Clinical manifestations suggesting anaerobic infections were common and three-quarters (n = 39) of 52 patients received adequate empirical antimicrobial treatment. Documentation of anaerobic bacteremia seldom influenced antimicrobial treatment. The 30-day mortality was 25%. Although univariate analysis revealed that underlying malignancy (p=0.003), leukopenia (p=0.044) and absence of fever (p=0.047) were associated with mortality, only malignancy (p=0.007) was an independent risk factor in the multivariate analysis. PMID- 16341346 TI - Etanercept therapy in children with juvenile rheumatoid arthritis. AB - Etanercept is an effective inhibitor of tumor necrosis factor that has shown a beneficial effect in patients with juvenile rheumatoid arthritis (JRA) that did not respond to other disease-modifying drugs. Here we report 3 patients with JRA who were refractory to traditional therapy; 1 with systemic JRA and 2 with polyarticular JRA. They received etanercept 0.4 mg/kg (maximum 25 mg) subcutaneously, twice a week for 3 months. The symptoms of arthritis improved significantly except that the patient with systemic JRA had disease flare-up during etanercept therapy. Two patients had upper respiratory tract infection during etanercept therapy and 1 suffered from seizure attack. The 2 patients with polyarticular JRA had disease flare-up within 2 months after etanercept was discontinued. This is the first report of etanercept treatment in JRA patients in Taiwan. PMID- 16341347 TI - Treatment with etanercept for patients with juvenile rheumatoid arthritis in Taiwan--a preliminary report. AB - Tumor necrosis factor (TNF) is a major inflammatory cytokine involved in the pathogenesis of juvenile rheumatoid arthritis (JRA). Etanercept, approved in the United States and in Europe for use in patients with rheumatoid arthritis (RA) and JRA, is an effective inhibitor of TNF that has been shown to provide rapid and sustained improvement in both diseases. Here we report the preliminary results of etanercept use in 3 cases of JRA with poor response to traditional therapy including non-steroidal anti-inflammatory drugs and disease-modifying antirheumatic drugs. Two of the patients had polyarticular JRA and 1 had systemic JRA. Etanercept was administered at a dosage of 0.4 mg/kg (maximum 25 mg) subcutaneously twice a week. Clinical as well as inflammatory parameter improvement was noted after use of etanercept in all cases. The preliminary results of etanercept use in these 3 cases showed significant clinical benefit without obvious adverse effects. PMID- 16341348 TI - Taiwan experience with etanercept in juvenile rheumatoid arthritis. PMID- 16341349 TI - Guidelines for the use of antimicrobial agents in patients with febrile neutropenia in Taiwan. PMID- 16341350 TI - [Enrique Fanta Nunez]. PMID- 16341351 TI - [Is it possible to eliminate mother-offspring transmission of human immunodeficiency virus? Main aspects of the new Standard for Prophylaxis of Vertical Transmission]. PMID- 16341352 TI - [Acinetobacter baumannii pandrug-resistant: update in epidemiological and antimicrobial managing issues]. AB - In the last two decades Acinetobacter baumannii has emerged as a major relevant world nosocomial pathogen. A. baumannii can be a causal agent of diseases like pneumonia, bacteremia, meningitis, soft tissue and urinary tract infections, associating to high mortality. Several national and foreign communications reveal the isolation of A. baumannii resistant to almost all commercially available antimicrobial agents, drastically limiting the therapeutic options. To optimize the therapy of these infections, the need of development of new antibacterial agents is raised as is the revival of certain forgotten compounds, as the polymyxins. In order to check and evaluate the information on the management of multi-resistant A. baumannii infections, we performed a systematical review of the medical scientific literature, that included Medline and LILACS, identifying and categorizing the clinical relevancy of the sources gathered to the date of this investigation. Clinically relevant epidemiological aspects, microbiological information and clinical studies in patients with pandrug (AB-PDR) or multidrug resistant (AB-MDR) A. baumannii infections were reviewed. The response adapted to the management of AB-PDR infection is complex, its eradication needs adherence to suitable practices of infection control and the prudent and effective use of antimicrobial therapy. Potential options of therapy could be colistin, betalactam associations with sulbactam and tetracyclines, but there are no random and controlled studies in the matter. PMID- 16341353 TI - [Detection of HIV infection in pregnant women by rapid testing: a successful strategy to reduce its vertical transmission]. AB - A high percentage of Uruguayan pregnant women are not under medical control. As a consequence, vertically transmission of HIV infection reaches to 50%. The aim of this study was to know the prevalence of HIV infection in pregnant women who did not know their serological status and to decrease mother-to-child transmission. PATIENTS AND METHODS: from January 2002 to January 2004 the HIV rapid test was performed to every pregnant woman that assisted to a public Uruguayan hospital unaware of her condition. The proper prophylactic decisions were adopted according to gestational age. The newborn infants were classified according to CDC criteria. HIV infection prevalence in pregnant women and in their newborn infants was calculated. RESULTS: there were 34,338 obstetric consultations and 4,599 rapid tests were performed. Fifty-nine turned out positive in 58 women, 8 of them knew their serological status previously. The HIV infection prevalence was 1.1% (IC95% 0.8-1.4). Five cases were discharged: 1 false positive and 3 miscarriages and 1 abortion. Ten women dropped out in the follow up. Twelve women received TARV during pregnancy for over a week. Thirty-nine infants could be controlled: 33 seroreverted, 4 were exposed and 2 became infected. Mother-to child transmission was 5.1%. If all patients who dropped out the follow up were infected, the transmission rate should be of 20.4%; therefore the infection would have been prevented in 16 children. As many women and children were lost, other complementary actions as counseling and social worker interview should be adopted in order to improve the yield of rapid test screening strategy. PMID- 16341354 TI - [Human immunodeficiency virus (HIV) infection in pregnancy: antiretroviral treatment (ART) and mode of delivery]. AB - Antiretroviral therapy (ART) administered to pregnant women infected with HIV diminishes the rate of perinatal viral transmission. This is true for mono-, bi-, or tri-therapy (HAART), with the greatest effects being seen in the latter case. Nevertheless, when these therapies are employed, potential risks to the mother and fetus must be considered. These risks include hyperglycemia, lactic acidosis, mitochondrial toxicity, cutaneous rash, hepatitis, hypertension, and premature labor. Elective caesarean section reduces the perinatal transmission of HIV in patients with or without monotherapy, but has not shown a benefit in patients on tri-therapy (HAART). This article reviews the evidence for and against antiretroviral therapy and elective caesarean section in the setting of HIV in pregnancy and proposes treatment guidelines for these patients. PMID- 16341355 TI - [Vaccination in premature infants: an issue many times forgotten]. AB - Vaccine-preventable diseases may be serious and even life threatening in premature infants. Opportunities for immunization are frequently missed due to parental or healthcare personnel misconceptions on safety or immunogenicity of vaccines in these children. In addition, the impact of infectious diseases is frequently underestimated in preterms. There is no evidence of a higher incidence rate of vaccine associated adverse reactions in premature babies. For the majority of preterm infants vaccines provide protection against severe disease except for those extremely premature (less than 1500 grs) in whom the immune response may be less. Adherence to the EPI Vaccine Program is critical in this population with an immunization schedule similar to that of term infants, following chronological age. Premature follow-up programs should include additional vaccines not included in the EPI Program, such as conjugated pneumococcal vaccines and annual influenza inactivated vaccine. An additional strategy to protect preterms is the strict adherence to vaccination schedule among contacts including use of special vaccines in older children and adult contacts such as influenza and acellular pertussis vaccines. PMID- 16341356 TI - [New rotavirus vaccines: a reality at last]. AB - Rotavirus is the most common cause of severe acute diarrhea among children less than 3 years of age. This infection causes 25 million medical visits, 2 million hospitalizations and 440,000 deaths every year worldwide. The development of rotavirus vaccines has followed a long and difficult path seriously hampered by the abrupt withdrawal of Rotashield in 1999 due to its association with intestinal intussusception. After six years of intense research, the world celebrates the licensing of two new rotavirus vaccines, different in formulation and administration schemes albeit clearly safe and not associated with intussusception as demonstrated by large Phase III trials including more than 60,000 children. These vaccines, Rotarix from Glaxo SmithKline Biologicals and Rotateq from Merck Sharp & Dohme, have demonstrated to be highly efficacious against severe rotavirus gastroenteritis caused by the most prevalent serotypes. Prompt incorporation of these new vaccines into the immunization programs of countries, especially those in the poorest areas of the world where these vaccines are most needed, will require joint efforts between governments, vaccine manufacturers, international and non-governmental organizations and foundations. PMID- 16341357 TI - [Arcanobacterium haemolyticum]. PMID- 16341358 TI - [Disseminated fusariosis by Fusarium oxysporum in an adult patient with acute myeloid leukemia and severe febrile neutropenia]. AB - We report the first case of disseminated fusariosis in an adult patient in Chile, with a hematologic neoplasia and chemotherapy that evolved with febrile prolonged neutropenia, refractory, treated with wide spectrum antibiotics and that developed a systemic infection with sinusal, cutaneous and pulmonary compromise by Fusarium oxysporum. He was refractory to antifungal treatment with amphotericin B deoxycholate and caspofungin that were administered in a sequential schedule. The fatal outcome of this patient was associated to persistent neutropenia and infection caused by a filamentous fungus usually resistant to antifungal therapy. PMID- 16341359 TI - [Endocarditis caused by Streptococcus pneumoniae in children: case report and review]. AB - Endocarditis caused by Streptococcus pneumoniae in children is an infrequent disease, corresponding to 3-7% of all cases of endocarditis. However, it is highly relevant because of its severity with the possibility of producing valvular ring abscesses and destruction, and high mortality that reaches up to 61% if medical and surgical treatment are not started early in the course of the illness. Over 50% of cases are associated to other sites of infection such as meningitis, pneumonia, sinusitis or mastoiditis. We report a 10-months-old infant who was admitted with meningitis and endocarditis due to S. pneumoniae, who presented with severe heart failure and required aortic valve replacement. A review of the literature of endocarditis caused by S. pneumoniae in pediatrics is presented. PMID- 16341360 TI - [Microbes aboard your Magesty's ships]. PMID- 16341361 TI - [Penicillin G use in pneumococcal pneumonia treatment]. PMID- 16341363 TI - [75th anniversary of the Pontifical Catholic University of Chile School of Medicine]. AB - The Pontifical Catholic University of Chile founded its School of Medicine in 1930, preceded by the Schools of Medicine of the University of Chile (founded in 1833) and the University of Concepcion (founded in 1924). The founders of the new School were prominent members of the Catholic Church, advised by distinguished Chilean physicians and scientists, most of them already full Professors at the University of Chile. During the first decades, only a small number of full-time faculties could be recruited and the academic team included mostly part-time teachers, generally shared with the already established Medical Schools, as well as practising physicians with little or no previous teaching experience. Along 75 years of fruitful existence, this Medical School has evolved to become one of the leading organizations in graduate and post-graduate medical education in Latin America, with high standards in teaching methodology and a superb productivity in clinical and biomedical research. The history of this Medical School stands as an example to the currently nascent Medical Schools founded by private universities in Chile. The standards of high quality medical education should be maintained for the well-being of our population so that these new Medical Schools can become respected and accepted as peers by the entire community of Chilean Universities. PMID- 16341364 TI - [Long-term results of repair versus replacement for degenerative mitral valve regurgitation]. AB - BACKGROUND: Mitral valve repair is considered better than mitral valve replacement for degenerative mitral regurgitation. AIM: To evaluate late clinical results of mitral valve repair as compared to mitral valve replacement in patients with degenerative mitral regurgitation. PATIENTS AND METHODS: All patients subjected to open heart surgery for degenerative mitral regurgitation between 1990 and 2002 were assessed for surgical mortality, late cardiac and overall mortality, reoperation, readmission to hospital, functional capacity and anticoagulant therapy. Eighty eight patients (48 males) had mitral valve repair and 28 (19 males) had mitral valve replacement (23 with a mechanical prosthesis). Mean age was 59.9 +/- 14.8 (SD) and 61.3 +/- 14.6 years, respectively. Sixty three percent of patients with repair and 50% of those with valve replacement were in functional class III or IV before surgery. RESULTS: Operative mortality was 2.3% for mitral valve repair and 3.6% for mitral valve replacement (NS). Also, there was no statistical difference in the need of reoperation during the follow-up period between both procedures (2.3% and 0%, respectively). Ninety four percent of the replacement patients but only 26% of the repair patients were in anticoagulant therapy at the end of the follow-up period (p < 0.001). Ten years survival rates were 82 +/- 6% for mitral valve repair and 54 +/- 11% for replacement. The corresponding cardiac related survival rates were 89 +/- 6% and 79 +/- 10%. At the end of follow-up, all surviving patients were in functional class I or II. Ten years freedom from cardiac event rates (death, cardiac related rehospitalization and reoperation) were 90 +/- 3% for mitral valve repair and 84 +/- 6% for replacement. CONCLUSION: Repair of the mitral valve offers a better overall survival and a better chance of freedom from cardiac events as well as need for anticoagulation 10 years after surgery. PMID- 16341365 TI - [Prevalence of cardiovascular risk factors among patients suffering vascular events on admission and one year later]. AB - BACKGROUND: International studies show a low compliance with norms for the management of cardiovascular risk factors. AIM: To assess the prevalence of risk factors in patients admitted for a coronary or vascular event and to evaluate the proportion of patients that normalize these factors after one year of follow up. MATERIAL AND METHODS: Three hundred and fifty seven patients aged 64+/-13 years (264 males), admitted to a University Clinical Hospital for a coronary or vascular event were studied. They were educated about cardiovascular risk factors and followed by their treating physicians for a mean of 11.9+/-2 months. During this period, smoking habits, body mass index. blood pressure, serum lipid levels, blood glucose and the appearance of new cardiovascular events were registered. RESULTS: One year survival was 96% (all 13 deaths were of cardiac origin). Eighty seven percent of patients were free of major cardiovascular events. At discharge from hospital and at the end of follow up 49% and 44% had a total cholesterol over 200 mg/dl respectively, 9,6% and 20,8% had systolic pressure over 140 mmHg. There was no diastolic hypertension in these patients, 27% and 31% had a body mass index over 25 kg/m2 and 2% smoked (versus 32% before the event). CONCLUSIONS: After one year of follow up, the prevalence of risk factors in patients that had suffered a cardiovascular event, continues to be high. PMID- 16341366 TI - [No association between persistence of the parasite and electrocardiographic evolution in treated patients with Chagas disease]. AB - BACKGROUND: At the present time the assessment of results of treatment of Chagas disease is mainly parasitological. Anti Trypanosoma cruzi IgGs remain positive practically lifelong and electrocardiographic tracings are not usually used as criteria of improvement. AIM: To determine, in a long term follow up, if electrocardiographic evolution is associated with the persistence of the parasite in treated patients with chronic Chagas disease. MATERIAL AND METHODS: Thirty patients with chronic Chagas disease that participated in a randomized trial of treatment with itraconazole or allopurinol, were studied. Seven years after treatment, patients were classified in group I if they had a positive xenodiagnosis test, polymerase chain reaction and hybridization in blood or in group II if they had negative tests. A 12 lead electrocardiogram (EKG) was performed each year to all patients. RESULTS: Seventeen patients were classified in group I and 13 in group II. At baseline 10 patients in group I and 8 in group II had a normal EKG. Six years after treatment 13 patients in group I and 10 in group II had a normal tracing. Of those with a normal tracing at baseline, only one patient in each group presented alterations after six years. A regression of abnormal tracings was observed in four and three patients of groups I and II respectively. CONCLUSIONS: There is no association between the persistence of the parasite in treated patients with Chagas disease and the evolution of electrocardiographic tracings. PMID- 16341367 TI - [Early and long term results of mechanical aortic valve replacement at the Instituto Nacional del Torax in Chile]. AB - BACKGROUND: Mechanical aortic valve replacement (AVR) results have been published extensively in industrialized countries. AIM: To assess our immediate and late results in patients subjected to AVR. PATIENTS AND METHODS: We retrospectively studied 194 patients subjected to isolated AVR between 1995 and 2003. Mean age was 57 +/- 13 years and 119 (61%) were male. One hundred thirty nine (73%) were in functional class III-IV, 20 (10%) had a previous cardiac operation and 25 (13%) were operated as an emergency. Surgical indication was stenosis in 110 (58%), regurgitation in 49 (26%) and stenosis/regurgitation in 31 (16%). Etiology was bicuspid valve 56 (29%), degenerative lesions 55 (28%), rheumatic valve disease 38 (20%) and endocarditis 27 (14%). Medtronic Hall was the most common prosthesis used in 157 patients (81%). Mean cardiopulmonary bypass time 97 +/- 29 min and mean cross clamp time was 69 +/- 21 min. RESULTS: Operative mortality was 4.6% (3% in elective surgery, 16% in emergency surgery and 0% in reoperations). Follow-up was complete in 100% of cases, totalizing 636 patients-year. Survival was 91 +/- 2%, 80 +/- 4% and 73 +/- 6%, at 1, 5 and 7 years, respectively. Multivariate risk analysis identified renal failure and endocarditis as predictors of early and late mortality. During follow up, the linear incidence rate for hemorrhage was 3.29% /patients-year, thromboembolism 2.04% patients-year and endocarditis 1.1% patients-year. CONCLUSIONS: AVR has low overall and elective mortality. Midterm survival is good but linear event rates related to anticoagulant treatment are higher than those previously published in industrialized countries. Renal failure and endocarditis were risk factors for early and late death. PMID- 16341368 TI - [Effects of TeenSTAR, an abstinence only sexual education program, on adolescent sexual behavior]. AB - BACKGROUND: Urgent measures are required to stop the increase in the frequency of pregnancies and sexually transmitted diseases among teenagers. A means of facing this problem is promoting sexual abstinence among youngsters. There are studies that confirm the efficacy of this approach. AIM: To show the results of the application of a holistic sexuality program (TeenSTAR) among Chilean teenagers. SUBJECTS AND METHODS: Students attending basic or high school were divided into a control or study group. The control group (342 students) received the usual education on sexuality given by their schools and the study group (398 students) participated in twelve TeenSTAR sessions lasting 1.5 hours each, given by a trained professor. Assessment of achievements was made using an anonymous questionnaire answered at the start and end of the program. RESULTS: The rates of sexual initiation among control and study groups were 15 and 6.5%, respectively. Among sexually active students, 20% of those in the study group and 9% of those in the control group discontinued sexual activity. CONCLUSIONS: A higher proportion of students in the TeenSTAR program retarded their sexual initiation or discontinued sexual activity and found more reasons to maintain sexual abstinence than control students. PMID- 16341369 TI - [Viral Tax protein expression in salivary glands of patients infected with human t-cell lymphotropic virus type I and Sicca Syndrome]. AB - BACKGROUND: Human T-cell lymphotropic virus type I (HTLV-I) is a retrovirus that influences cellular metabolism modifying biological responses. This results in oncogenic, degenerative or inflammatory changes. The myelopathy associated to HTLV-I or tropical spastic paraparesia (HAM/TSP) is a mainly degenerative response to the virus infection. On the other hand, Sjogren syndrome has an inflammatory appearance. The immunohistochemical study of CD-4, CD-8 and CD45 lymphocytes, metalloproteinase MMP-9 and viral Tax protein in pathological samples of salivary glands may help to differentiate primary from viral Sicca syndrome. AIM: To perform an immunohistochemical study of salivary glands of patients with HAM/TSP and Sicca syndrome and control subjects. MATERIAL AND METHODS: Pathological samples of salivary glands from 53 patients with HAM/TSP and Sicca syndrome and 10 control subjects, were studied. Immunohistochemistry was performed using antibodies against CD-4, CD-8 and CD-45 lymphocytes, metalloproteinase MMP-9 and viral Tax protein. RESULTS: Only in patients with HAM/TSP and Sicca syndrome, the presence of Tax protein was observed in CD-4 and CD-8 lymphocytes and in glandular acini. CONCLUSIONS: Patients infected with HTLV I express Tax protein in salivary glands. This finding has diagnostic and pathogenic implications. PMID- 16341370 TI - [Influence of personality and learning styles in the choice of medical specialty]. AB - BACKGROUND: Several studies indicate that doctors who work in the same area of the medical profession tend to behave somehow similarly. Thus, it has been suggested that personality relates to the medical specialty choice. However, it is not known whether people self-select into the medical specialties according to their personality or the professional practice in a particular field influences their behavior. AIM: To explore the possible association between the graduate's personality features and learning styles and their chosen specialty. SUBJECTS AND METHODS: The psychological preferences and learning styles of 65 students of the 2001-graduating cohort of the Pontificia Universidad Catolica de Chile School of Medicine were evaluated with the Myers Briggs Type Indicator and the Kolb Learning Style Inventory, respectively. These variables were correlated with the information of their specialty choice or occupation two years after graduation. RESULTS: Graduates distributed unevenly in different areas of the medical profession. Surgical specialties concentrated a larger proportion of extraverted, intuitive and structured doctors, whereas in Pediatrics and Internal Medicine predominated intuitive and people-oriented MD's. Primary Care concentrated individuals with introverted, intuitive and flexible attitudes. Convergent learners (interested in problem-solving) preferred Surgery and Primary Care whereas Assimilator learners (abstract-reflexive) chose more frequently Internal Medicine, Pediatrics and Psychiatry. CONCLUSIONS: According to their personality and learning style, graduates tend to self-select into different medical specialties. This information may help medical graduates to guide their specialty choice process, and medical educators to develop learning experiences that take into account the individual differences of their residents. PMID- 16341371 TI - [Prevalence of antibiotic resistant Enterococcus spp in waste waters in the north of Chile]. AB - BACKGROUND: There is little information available in Chile on the distribution of Enterococcus spp in waste water and its implications in transmission of antibiotic resistance through the water cycle. Enterococcus spp are common in nosocomial infections and may spread antibiotic resistance through the food chain. AIM: To determine the presence of antibiotic resistant Enterococcus spp in the sewage of Antofagasta, Chile. MATERIAL AND METHODS: Samples of sewage from two sewage treatment plants and from the Public Hospital of Antofagasta collector were obtained. Enterococcus spp were isolated on m-Enterococcus agar containing ampicillin, vancomycin and streptomycin. The isolates were identified and subjected to biochemical typing (PhPlate). Minimal inhibitory concentration determination was performed by agar dilution technique. RESULTS: High counts of resistant Enterococcus spp were found on the streptomycin plates, lower on ampicillin and very low on vancomycin plates. A total of 63 Enterococcus spp strains were typed and the identification showed 5 different species; E faecalis (65%), E faecium (14%), E hirae (13%), E durans (6%) and E gallinarum (2%). The typing revealed a high diversity among the isolates. Two biochemical phenotypes were predominant, C1 (21 strains) and C6 (7 strains). Both were highly resistant to gentamycin and streptomycin; moderately resistant to ampicillin, chloramphenicol, tetracycline and ciprofloxacin, and with intermediate susceptibility to vancomycin. Both phenotypes were found in the sewage of the hospital collector and in the treatment plants. CONCLUSIONS: In the sewage of Antofagasta we found dominating phenotypes of multiresistant Enterococcus spp. Sewage could be an important way of transmission of these microorganisms. PMID- 16341372 TI - [Clinical and functional benefits of adding theophylline to a standard treatment with short acting bronchodilators in patients with COPD]. AB - BACKGROUND: Although theophylline is considered a third line bronchodilator drug for the treatment of chronic obstructive pulmonary disease (COPD), it is widely used in Chile, because it is administered orally and has a moderate cost. AIM: To evaluate if theophylline adds clinical and/or functional benefits when associated to standard recommended inhaled bronchodilator therapy. SUBJECTS AND METHODS: Thirty-eight stable COPD patients who accepted to participate in the study approved by the Ethics Committee of our institution were studied. Using a randomized double-blind placebo-controlled study, theophylline (250 mg) or placebo was administered twice a day for 15 days in addition to inhaled salbutamol and ipratropium bromide. Prior to and at the end of the study, patients underwent: a) a spirometry to evaluate changes in dynamic pulmonary hyperinflation using slow vital capacity (SVC) and inspiratory capacity (IC), b) the 6 min walking distance (6 MWD); and c) measurement of maximal inspiratory and expiratory pressures. Dyspnea and quality of life (QoL) were evaluated using appropriate questionnaires. RESULTS: Compared to placebo, patients on theophylline showed significant increases in SVC (p=0.014), IC (p=0.002), and 6 MWD (p=0.005). They also experienced an improvement in dyspnea (p=0.042) and QoL (p=0.011). All patients improved at least one of these parameters with 53% of the patients showing an improvement in 3 or more. CONCLUSIONS: Our results indicate that adding theophylline to standard treatment with inhaled bronchodilators provides additional benefits in stable COPD patients by reducing dynamic pulmonary hyperinflation, improving exercise tolerance, dyspnea and QoL. PMID- 16341373 TI - [Sirolimus for rescue of recurrent acute rejection and diabetes mellitus after liver transplantation: report of one case]. AB - Sirolimus (SRL) is a new immunosuppressive drug approved for renal transplantation, but is being used increasingly in orthotopic liver transplantation (OLT). Compared with the calcineurin inhibitors, SRL has different mechanisms of action and side effects profile. Thus, this drug offers significant potential advantages over other immunosuppressive agents. SRL does not cause glucose intolerance, hypertension or renal failure, but it may cause dyslipidemia, hepatic artery thrombosis, thrombocytopenia, anemia, leukopenia, oral mucosa ulcers, edema, arthralgias and wound complications. SRL inhibits the signal of interleukin 2 at a post-receptor level, inhibiting lymphocyte proliferation and fibroblast proliferation. It also has antineoplastic and antifungal effects. We report a 10 years old girl who underwent OLT, experiencing a biopsy-proven recurrent acute rejection (AR) in spite of using three immunosuppressive agents (tacrolimus, mofetil micofenolate and steroids). She developed diabetes mellitus as a consequence of the immunosuppressive therapy. She was rescued with SRL, not experiencing AR again. Mofetil micofenolate, steroids and insulin could be discontinued and tacrolimus doses were reduced, without experiencing severe complications. SRL is a new and safe immunosuppressive agent for rescue in patients with OLT and recurrent AR. PMID- 16341374 TI - [Malignant neuroleptic syndrome and polyserositis associated to clozapine use: report of one case]. AB - Malignant neuroleptic syndrome is a complication of antipsychotic medication use. Clozapine use is also associated with polyserositis and eosinophilia. We report a 17 years old female treated with clozapine, valproic acid, lithium carbonate and lorazepam that consulted in the emergency room for confusion, lethargy, catatonia, rigidity, myalgya and fever. Complete blood count showed eosinophilia. An abdominal CAT scan showed ascites and pleural effusion. Clozapine was discontinued and bromocriptine was started. One week after admission, the patient remained febrile and liver enzymes were elevated. Valproic acid was discontinued. Inflammatory parameters stated to subside and the patient was discharged afebrile days after admission. PMID- 16341375 TI - [Seventy five years of the Medical School of the Pontifical Catholic University of Chile]. AB - Aiming to join academic excellence and an ethical and Christian approach to medical profession, the Medical School of the Pontifical Catholic University of Chile initiated its activities in 1930. Since then, the associated Health Care Network has incorporated all the technological breakthroughs in medicine and developed all the specialties. Undergraduate teaching is oriented to promote creativity and innovation. There is also a special concern about humanity of Medicine, throught the Program of Humanistic Medical Studies and the Bioethics Center. Post graduate education is also an important activity of the School, through specialty training, Master and Doctorate programs. Researchers have also obtained important grants and generated a great number of publications in high impact journals. Our University is defined as "complex", meaning that we must take important challenges, be creative and lead knowledge generation. We must also improve ourselves to serve in the best possible way our students and the Country. Paraphrasing the words of our founder, Monsignor Carlos Casanueva, we must train physicians that will serve our community not only with science but also with humanity. PMID- 16341376 TI - [An update on esophageal perforation]. AB - Esophageal perforation is a complicated clinical entity that demands a high level of diagnostic and therapeutic skills. The management alternatives vary from conservative treatment to esophagectomy, including primary suture and esophageal exclusion. This paper is a review of the literature and personal experience with this condition, focusing on etiology, clinical presentation, diagnostic workout, treatment, complications and mortality. Finally, we propose a management algorithm. PMID- 16341377 TI - [An update on the treatment of psychological and behavioral symptoms associated to dementia]. AB - Behavioral disturbances in dementias are relevant because they are very common, they worsen patients' medical and cognitive condition, increase the caregiver burden and accelerate the admission to nursing homes. The different behavioral disturbances in dementias can appear isolated, but in most cases, patients present a combination of disturbances producing one of two main syndromes: the psychotic or the affective syndrome. Proper handling of these behavioral disturbances requires a correct characterization of the syndrome, discarding medical or environmental causes and selecting the drug with the better effectiveness and safety profile for each patient. In spite of the important number of psychotropics available, there is not enough scientific evidence about their real effectiveness and security in patients with Alzheimer's disease. This article reviews recent advances in the treatment of most common and disruptive behavioral disturbances. PMID- 16341378 TI - [Reflections on evidence-based medicine]. AB - This is a review of the basic concepts of evidence, medicine, evidence-based medicine and its effects. Evidence-based medicine is a contribution for statistical design and management, that allows the gathering of scientific information, but does not completely exclude bias. Its methodology proceeds with order and caution, generating a highly probable mass of knowledge. The quality of the information obtained from diverse sources such as meta-analysis to expert opinions is classified in different levels according to the "strength" of the evidence in which it is based. The best contribution of evidence-based medicine is probably for diseases of incidence and does not replace clinical experience and patients-physician relationship. Basically we treat ill people and not illnesses. Without clinical experience, medical practice can be tyrannized by evidence-based medicine that can become inapplicable or inappropriate. Without evidence-based medicine, medical practice will not be updated, limiting our professional skills. PMID- 16341379 TI - [Cervantes, Don Quixote and medicine]. PMID- 16341380 TI - [Clinical and molecular features of cardial gastric cancer associated to Epstein Barr virus]. AB - BACKGROUND: Mortality caused by cardial gastric cancer in Chile, is increasing. Previously we demonstrated an association between Epstein Barr virus and this specific location of gastric cancer. AIM: To perform a clinical and molecular characterization of cardial gastric cancer associated to Epstein Barr virus. MATERIAL AND METHODS: Epstein Barr virus was identified in 93 cardial gastric tumors, by in situ hybridization. Clinical and pathological features, survival and expression of p53 and c-erbB2 were compared between tumors with or without the presence of the virus. RESULTS: Twenty two (23.6%) tumors expressed Epstein Barr virus. No difference in sex or age of patients with tumors positive or negative for the virus was observed. Epstein Barr positive tumors had a tendency to have a higher frequency of Bormann III endoscopic appearance and a lower frequency of p53 accumulation (p=0.06). Five years survival was 67% and 42% of tumors positive and negative for the presence of the virus, respectively (p=0.57). CONCLUSIONS: Our results, although not significant, show a tendency towards unique characteristics of cardial gastric tumors associated to Epstein Barr. PMID- 16341381 TI - [Comparison of cardiac output measurement in critical patients using transesophageal Doppler or thermodilution: a preliminary report]. AB - BACKGROUND: Cardiac output can be measured non invasively by transesophageal Doppler. This is an alternative to measure it by thermodilution with a catheter in the pulmonary artery. AIM: To compare both methods of cardiac output measurement. MATERIAL AND METHODS: Simultaneous measurement of cardiac output by transesophageal Doppler and thermodilution with a catheter in the pulmonary artery in four male critical patients, aged 60+/-12 years, hospitalized in a University Hospital. The Bland and Altman method to compare the concordance between two measurements, was used. RESULTS: Forty measurements were performed. The results of both methods had a correlation coefficient of 0.98. According to the Bland and Altman method, the difference between both methods was -0.5 L with a precision of 0.52 L/min (95% confidence interval -1.51 to 0.52 L/min). Considering that a change between two sequential measurements is considered significant when the difference is more than 15%, both measurements agreed in 83% of cases, that there was a change in cardiac output. CONCLUSIONS: Transesophageal Doppler is a promising non invasive technique to measure cardiac output in critical care patients. It becomes a valid alternative to the thermodilution technique. This preliminary experience must be confirmed in a larger series. PMID- 16341382 TI - [Screening of the delta-F508 mutation and analysis of two Single Nucleotide Polymorphism of the CFTR gene, in a sample of the general population of Valparaiso, Chile]. AB - BACKGROUND: The Cystic Fibrosis (CF) carrier rate in Chile was estimated to be 1/40. CF is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. Delta F508 mutation is the most common in CF patients in Chile and worldwide. Delta F508 has linkage disequilibrium with two Single Nucleotide Polymorphisms (SNP), often used to define the haplotypic frameworks of CF mutations. AIM: To know the frequency of the delta F508 mutation and to establish the SNPs, M470V and T854T, haplotypic frequency, in a Valparaiso general population sample. SUBJECTS AND METHODS: Fifty subjects were studied. Genetic material was obtained from blood samples, amplified by PCR and analyzed by restriction fragment length polymorphism. RESULTS: Two of the 100 chromosomes analyzed, carried the delta F508 mutation. Therefore, the observed frequency carrier rate (0.02) was higher than the expected (0.01). Both carrier chromosomes had the same SNPs haplotypic framework (1-2). In normal chromosomes, the haplotype 2-1 was the most common. DISCUSSION: These results suggest that the chromosomes that bear delta F508 mutation have most likely a Mediterranean European origin, since this haplotypic framework has been reported in that region. We suggest that CF could be more common in Valparaiso than it was previously. PMID- 16341383 TI - [Primary sclerosing cholangitis: a twelve-year experience]. AB - BACKGROUND: Primary sclerosing cholangitis (PSC) is a chronic cholestatic disorder characterized by progressive inflammation and fibrosis of the biliary tract, evolving to cirrhosis. It is commonly associated with inflammatory bowel disease (IBD). AIM: To communicate the clinical characteristics of patient with PSC seen in two reference centers. PATIENTS AND METHODS: Review of medical records of patients with PSC confirmed by liver biopsies. The clinical picture, laboratory abnormalities, imaging studies and treatment were recorded. RESULTS: Thirty three patients (aged 16 to 73 years, 64% female) were identified. They corresponded to 1.7% of liver biopsies done between 1991-2003. Clinical presentation was a cholestatic picture in 40%, right upper abdominal pain in 36%, a dysenteric syndrome in 9% and varied symptoms in 15%. Laboratory tests showed cholestasis in 94% and positive anti ANCA, SMA, ANA and AMA antibodies in 28, 18, 15 and 9% of cases, respectively. Endoscopic retrograde cholangiopancreatography and magnetic resonance cholangiography were diagnostic in 43 and 58% of patients, respectively. There was an association with ulcerative colitis in 12% of cases. Liver biopsies showed grade I PSC in 76% and grade II-III in 6% of patients. It also showed a concomitant chronic hepatitis and primary biliary cirrhosis in 12 and 6% of cases, respectively. Treatment consisted on ursodeoxycholic acid (UDCA) in 45%, UDCA plus 5-aminosalicylic acid derivatives in 12% and UDCA plus immunosuppresors in 12% of patients. Two patients had to be transplanted. CONCLUSIONS: PSC is an uncommon cause of chronic liver disease. It is suspected in cholestatic patients and confirmed with a liver biopsy. It can be associated with other autoimmune hepatic and extrahepatic diseases. PMID- 16341385 TI - [Diagnostic profiles at a Center for Memory Disturbances]. AB - BACKGROUND: The Memory Disturbances Center is a multidisciplinary team devoted to the study and treatment of patients with memory problems. AIM: To report the clinical features of patients consulting to this center. MATERIAL AND METHODS: Retrospective review of the clinical records of the first 100 patients consulting in the center. RESULTS: The age of consulting patients ranged from 17 to 93 years and 57 were female. The most common diagnoses were Alzheimer disease, depressive pseudo dementia, mild cognitive impairment, vascular dementia and adult attention deficit disorder. A formal neuropsychological assessment was performed in 60% and 20% entered a cognitive stimulation program. CONCLUSIONS: Although Alzheimer disease is the most common cause of memory disorders in the adult, concealed depression and attention deficit disorders are also common. A multidisciplinary approach allows a better diagnosis and treatment. PMID- 16341384 TI - [Hemolytic uremic syndrome: the experience of a pediatric center]. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) is one of the main causes of acute renal failure in the Chilean pediatric population. AIM: To report the features of patients with HUS, admitted to the pediatric ward of a clinical hospital. MATERIAL AND METHODS: Retrospective review of medical records of patients admitted with the diagnosis of HUS between 1995 and 2002. RESULTS: During the period, 58 patients were admitted with the diagnosis of HUS but only 43 (age range 1 month to 6 years, 22 females) had complete medical records for review. Ninety five percent presented with prodromic diarrhea, mainly dysenteric. Antibiotics were administered to 70%, in the previous days. Acute renal replacement, mainly peritoneal dialysis, was required in 40%. The clinical signs and laboratory parameters that correlated better with the indication for dialysis were anuria, hypertension, initial and permanently high serum creatinine and blood urea nitrogen. Four patients with blood urea nitrogen over 100 mg/dl but without anuria or hyperkalemia, were treated conservatively, and experienced an uneventful course (permissive azotemia). Hospital stay was almost 3 times greater in dialyzed than in non dialyzed children. No deaths related to HUS were reported in the study period. In an average follow up of 54 months, 11.6% of the patients developed chronic renal failure of diverse magnitude. CONCLUSIONS: Despite the fact that our study group behaved clinically similar to published HUS patients in other series, no mortality was observed in a retrospective analysis of patients with this disease. PMID- 16341386 TI - [Cardiovascular risk and metabolic profile in obese children and adolescents with low insulin sensitivity]. AB - BACKGROUND: Infantile obesity is associated with metabolic disturbances (hyperinsulinism, impaired glucose, dyslipidemia) that determine a higher risk of type 2 diabetes, high blood pressure and atherosclerotic vascular disease in adulthood. Insulin resistance is a central mechanism of complications of obesity and is associated to body fat mass. AIM: To investigate insulin sensitivity and its association with anthropometric and metabolic variables in obese children. PATIENTS AND METHODS: We studied 314 children aged 6 to 15 years. Of these, 235 had a body mass index (BMI) over the 95 percentile for age and sex (classified as obese) and 79 had a normal body mass index; 161 were pre-pubescent and 153 were pubescent. Weight, height, percentage of total body fat (% TBF) using the sum of 4 skinfolds, blood pressure (BP), waist circumference (WC), basal glucose (Glu) and insulin (Ins) were measured. Insulin sensitivity (IS) was calculated with the homeostasis model assessment (HOMA) and QUICKI index. Serum lipids, postprandial Glu and Ins were measured only in obese children. RESULTS: BMI, WC and % TBF had an inverse and significant correlation with basal IS (p <0.001). Obese children had higher BP, basal Ins, and HOMA and a lower QUICKI index, compared to normal weight children. A low basal IS was present in 58% of obese children. Obese children with low IS had higher WC, % TBF, triglycerides, total/HDL-cholesterol ratio, basal and 2-h post glucose Gli and Ins and lower HDL-cholesterol than obese children with normal IS. In prepubescent children, the risk of a low IS was 2.43 times higher with a TBF over 33%. In pubescent children, it was 2.92 times higher with a TBF over 37%. CONCLUSIONS: Low IS in prepubescent and pubescent obese children is associated with central obesity and a higher cardiovascular. PMID- 16341387 TI - [Training and research in forensic medicine: present situation and future challenges for medical schools in Chile]. AB - BACKGROUND: Lawyers need some medical knowledge and physicians must know about forensics. AIM: To explore training and research programs in forensic medicine in Chilean universities. MATERIAL AND METHODS: Deans of all Medicine Faculties in Chile were contacted by e-mail and invited to answer a questionnaire containing 21 questions. A survey of Chilean publications on forensic medicine was performed in Medline, Lilacs and SciELO databases. RESULTS: Fourteen deans answered the questionnaire. In all the responding faculties, forensic medicine is an obligatory course, generally during the fifth year and mostly combining theory with practice. In seven faculties, forensic medicine concepts are included in other courses. Forensics is taught in only two of 10 dental schools, two of 17 nursing schools, one of nine midwives schools and one of nine medical technology schools. It is not taught in phonoaudiology, kinesiology and nutrition schools. There are 74 physicians that teach the specialty but only 10 are certified by the National Board of Medical Specialty Certification (CONACEM). Treatment of most topics on forensics is insufficient. Thanatology is the strongest topic and forensic dentistry is the weakest. There are 52 publications in the area, mostly on "medical law". CONCLUSIONS: Forensic medicine is taught in medical schools mostly as thanatology. The knowledge of forensics among medical students is limited and must be improved. PMID- 16341388 TI - [Acquired von Willebrand disease as an unusual manifestation of primary hypothyroidism: report of two cases]. AB - Patients with hypothyroidism may have a minor increase in their bleeding tendency, causing easy bruising and menorrhagia. There is a positive correlation between factor VIII coagulant activity and thyroxin serum levels. Thus, patients with hypothyroidism have an acquired coagulation defect that is reversible with thyroxin supplementation. We report two sisters, aged 13 and 11 years, who met the criteria for von Willebrand's disease at the age of eight. Both sisters had a primary hypothyroidism due to Hashimoto's thyroiditis, diagnosed three years later. Thyroid hormone replacement normalized von Willebrand factor, factor VIII, and the bleeding diathesis disappeared. Acquired von Willebrand's disease is an unusual manifestation of hypothyroidism. However the possibility of hypothyroidism should be considered in patients presenting with von Willebrand disease. PMID- 16341389 TI - [Addition of an arterio-venous shunt during veno-arterial extracorporeal life support in a patient with Hantavirus pulmonary syndrome]. AB - A subgroup of patients infected with the Hantavirus develops a pulmonary syndrome (HPS) characterized by severe acute respiratory failure and myocardial depression, that has a high mortality rate. Extracorporeal life support (ECLS) could be a valuable therapeutic tool in such patients. We report a 24 years old male with HPS that was successfully managed when an arterio-venous shunt was added to a conventional veno-arterial ECLS technique. Precise criteria have been developed to predict which patients should be considered for this treatment. PMID- 16341390 TI - [HDL cholesterol: a new target in the treatment of lipid disorders and atherosclerosis?]. AB - Low plasma HDL cholesterol levels are an independent risk factor for atherosclerotic cardiovascular disease. During the past years, the study of HDL has been stimulated by the discovery of novel genes and proteins, which have provided new insights into the molecular and cellular mechanisms involved in HDL metabolism. In addition, recent clinical studies focused on treating low HDL as primary objective in high cardiovascular risk patients have been very encouraging. Furthermore, new drugs for raising HDL cholesterol are under development. This article reviews the recent progress in the HDL field and its important implications for the pathophysiology and treatment of atherosclerosis. In the near future, we expect that new drugs with specific and beneficial effects on HDL metabolism and the associated cardiovascular risk should improve the current approach to this disease with high burden on health systems and society. PMID- 16341391 TI - [Globalization of higher education in Europe. The precursor of changes in Chilean medical education]. AB - At the present time, higher education is a central process shaping the society of knowledge. The mutual strength of society and education influence worldwide changes. Globalization, that mainly embraces politics and commerce, is also promoting changes in professional training, modifying programs, structures and the organization of superior education. The European example is attractive and has a great projection. It will cause a profound impact in culture, development and life of Europeans. Universities in Latin America, in an attempt to achieve the same objectives than their European counterparts, are experiencing premonitory changes, that will have the same effects as in Europe. In the medical area, several changes will occur and we will have to provide initiatives to facilitate these changes in a timely manner. The revision of the model in its inception, is convenient to assimilate changes in our culture. PMID- 16341392 TI - [Bioethical issues involved in the prescription of levonorgestrel]. AB - The use of levonorgestrel for emergency post coital contraception after rape, has raised strong and recurring discussions during 2004 and 2005 in Chile. The debate has been centered in its presumed post fertilization or anti implantation effect, that some consider an abortive action. There are no scientific evidences supporting this effect, with divergences about the ontological status of the embryo. Therefore, the use of levonorgestrel implies bioethical decisions that, in a democratic and pluralistic society, should be solved considering individual and collective responsibilities, conditions of equity and the informed autonomy of the affected women. Their moral values and their capacity to assume the consequences of an assault on their dignity, honor and self-esteem, in addition to physical and mental injuries, should also be considered. PMID- 16341393 TI - [Molecular diagnosis of Cryptosporidium spp]. PMID- 16341394 TI - [Assessment of clinical trial quality and its impact on meta-analyses]. AB - OBJECTIVE: To evaluate whether different quality assessment tools applied to a group of clinical trials could be correlated, and what would be their impact on meta-analysis results. METHODS: Thirty-eight randomized controlled clinical trials were analyzed. These had been selected for a systematic review of the therapeutic efficacy of alpha interferon for treating chronic hepatitis B. The following tools were utilized: Maastricht (M), Delphi (D), Jadad (J) and the Cochrane Collaboration (CC) method (gold standard). The Spearman correlation coefficient was used to compare the results from the three methods. The Kappa test was used to assess the concordance between the reviewers in applying the tools, and the weighted Kappa test was applied to compare the quality ranking determined by the tools. The outcomes assessed in the meta-analyses were clearance of HBV-DNA and HBeAg. RESULTS: The studies presented regular to low quality. The concordance between reviewers varied according to the instrument utilized: D=0.12; J=0.29; M=0.33; and CC=0.53. The correlation was moderate and homogeneous (D/J=0.51; D/M=0.53; and J/M=0.52). The meta-analysis result relating to HBV-DNA ranged from RR=0.71 (95% CI: 0.66-0.77) to RR=0.67 (95% CI: 0.58 0.79). For HBeAg, the results ranged from RR=0.85 (95% CI: 0.80-0.90) to RR=0.85 ( 95% CI: 0.77-0.93). These results depended on the quality of the studies included. CONCLUSIONS: The quality assessment tools presented good correlation. In systematic reviews with the same direction of effect, the quality assessment may not significantly change the results. The Cochrane Collaboration method was the most reproducible method and easiest to apply. PMID- 16341395 TI - [Public care for breast cancer women in the state of Rio de Janeiro, Brazil]. AB - OBJECTIVE: New policies on oncology care implemented in the Brazilian Unified Health System (SUS) in November 1999 established substantial changes to cancer treatment in affiliate units. The objective was to describe the profile of these cancer services and their users after the implementation of new policies. METHODS: A descriptive study was carried out on breast cancer care in health services covered by SUS in the state of Rio de Janeiro, from 1999 to 2002. Data was collected in the healthcare units from outpatient registries and from the patients' high-complexity cancer procedure authorizations and their medical records. A random sample of 310 medical records of breast cancer women treated in 15 cancer health units was analyzed. Data analysis was performed using the percentile distribution of data according to categories of interest and the Chi square test to assess the association between variables. RESULTS: The study results show the predominance of breast cancer care in highly complex units (81.3%); public units (73.5%); and in the capital of the state (78.1%). There was also seen an imbalance among health units regarding patient attendance, where only one of them served 70% of women sampled. The use of interventions varied between patients with and without health insurance in lower complexity health services; non-insured patients were associated to lower use of interventions. It was also found underutilization of recommended interventions as well as utilization of contraindicated interventions. The characterization of population studied showed that 43.9% of the women were given a cancer diagnosis with no perspective of cure and 68.4% lived in cities having SUS-affiliated services. CONCLUSIONS: These results indicate relevant differences among health units and point out to the need of practical recommendations to the Brazilian national policy for cancer control. PMID- 16341396 TI - [Assessment of completeness of death reporting in Brazilian states for the year 2000]. AB - OBJECTIVE: To estimate and evaluate the completeness of adult death reporting for all Brazilian states. METHODS: Death statistics from the Brazilian Institute of Geography and Statistics (IBGE) and the Brazilian Ministry of Health for the period between 1999 and 2001 were compared by sex. Three techniques were used to evaluate the extent of death underreporting. The final estimate followed previously set criteria, resulting in four evaluation categories. RESULTS: For the first time, the Brazilian Institute of Geography and Statistics reported fewer deaths than the Ministry of Health. Death reporting showed to be at least "satisfactory" in all states in the South, Southeast, Mid-West and part of the Northeast region. The remaining states, from the Northeastern state of Piaui toward the North, were classified as "average" except for the state of Roraima. Fuller death reporting was seen among males. CONCLUSIONS: Increased death reporting was seen in all regions, particularly for those in North and Northeast regions. If this trend will continue, all Brazilian states will likely exceed 80% reporting by the year 2010. PMID- 16341397 TI - [Estimation of the dog and cat population in the State of Sao Paulo]. AB - OBJECTIVE: To estimate the total dog and cat population with owners in order to enable better planning of the control actions against diseases involving these animals. METHODS: The study was carried out in the non-metropolitan regions of the State of Sao Paulo, Brazil, from May to December 2002. Forty-one municipalities and 100 census tracts were surveyed. These were selected by probabilistic stratified cluster sampling in two stages. The strata were formed by grouping the municipalities according to their populations and living conditions. The Pasteur Sao Paulo Technique was used to obtain data on the canine population. This technique was developed to estimate and classify dogs according to their degree of dependence and restriction. RESULTS: Almost 53% of the 20,958 households visited owned a dog, and the average was 1.6 dogs per household. A total of 4,624 cats were found, concentrated in 12.6% of the households. The results indicate a dog/inhabitant ratio of 1:4.0 and a cat/inhabitant ratio of 1:16.4. CONCLUSIONS: The animal/inhabitant ratios were much higher than expected. Incorporation of these ratios into evaluations of the vaccination campaign against rabies has revealed more realistic coverage patterns, thus leading to renewed discussion of the vaccination targets for municipalities. An association was observed between the size of the municipality or its inhabitant's living conditions and the level of restrictions on dogs. PMID- 16341398 TI - [Frequent return as a novel strategy for tuberculosis treatment adherence]. AB - OBJECTIVE: Treatment dropout is one of the major obstacles to the control of tuberculosis. The aim of the present study was to evaluate the frequent return strategy instituted at the Tuberculosis Control Program, as well as the program as a whole, identifying and quantifying potential risk factors for dropout. METHODS: The present study was conducted in Ubatuba, Southeastern Brazil. All patients (n=244) notified by the Tuberculosis Control Program in the two years preceding and the two years following the introduction of the frequent return strategy in 1999 were included. We carried out a descriptive analysis of the data and analyzed treatment results and program evaluation indicators. We performed univariate and multivariate analysis in order to identify potential risk factors associated with dropout. We also analyzed the frequent return strategy using chi squared tests. RESULTS: The strategy reduced the risk of treatment dropout from 12.3% to 4.9%. Risk factors associated with dropout were lack of schooling (OR=3.01; p=0.051), unemployment (OR=3.36; p=0.046), and alcohol dependence (OR=3.56; p=0.014). CONCLUSIONS: The frequent return strategy reduced the risk of treatment dropout, although results did not reach statistical significance due to the small number of dropouts. This strategy may be an alternative for the supervised treatment for all patients recommended by the World Health Organization. PMID- 16341399 TI - [Validation of Diabetes Mellitus Knowledge (DKN-A) and Attitude (ATT-19) Questionnaires]. AB - OBJECTIVE: To present the cross-cultural adaptation of the Diabetes Knowledge Scale and Attitudes Questionnaires targeted to evaluate Brazilian Diabetes Mellitus patients. METHODS: These questionnaires underwent the following steps: presentation, translation, back translation, semantic and idiomatic assessments, cultural and conceptual similarities and a pilot test. They were administered in two opportunities a month apart in a sample of 61 Diabetes Mellitus type-2 patients from a university hospital. The study design included a test-retest reliability of the answers, which were analyzed and estimated by means of the Kappa coefficient. RESULTS: The findings suggested an adequacy of the instruments to the Portuguese language and Brazilian cultural identity. The Kappa coefficient in the reliability analysis showed levels of concordance from moderate to high (0.44 to 0.69) for most questions. CONCLUSIONS: The questionnaires proved to be of very easy understanding for the studied subjects and reliable and valid for use in the evaluation of diabetes educational programs in the reality of Brazil. PMID- 16341400 TI - [Validity of the Portuguese version of Clinical Dementia Rating]. AB - OBJECTIVE: To evaluate the validity of the Portuguese version of the Clinical Dementia Rating for classifying the cognitive function among the elderly. METHODS: The Mini Mental State Examination was utilized as a screening method for cognitive deficit among a cohort of 424 elderly. All those who scored <26 points (108 elderly) and 48 elderly with scores > or =26 were included in the study. The 156 subjects selected were submitted to clinical evaluation and neuropsychological tests for the diagnosis of dementia. Afterwards, both cases and non-cases were classified, according to the Portuguese version of the Clinical Dementia Rating, in the categories normal, borderline, mild, moderate and severe dementia. RESULTS: Among the 156 subjects selected, 122 were non cases, 62 (51%) were classified as normal (CDR=0) and 60 (49%) as borderline (CDR=0.5). Among the 34 cases of dementia, 17 (50%) were classified as mild dementia (CDR=1), eight (23%) as moderate (CDR=2) and six (18%) as severe dementia (CDR=3). Only three (9%) of the cases were considered borderline cases by the Clinical Dementia Rating. Its sensibility was 91.2% and the specificity was 100%. The positive predictive value was 100% and the negative predictive value was 97.6%. The Mini Mental State Examination scores declined significantly according to the degree of dementia. CONCLUSIONS: The Portuguese version of the Clinical Dementia Rating is a valid instrument for classifying the dementia status of the elderly. Almost half the cases considered normal by the diagnostic criteria of the Mini Mental State Examination were borderline cases according to the Clinical Dementia Rating and might correspond to cases of mild cognitive impairment, with an increased risk of conversion to dementia cases. PMID- 16341401 TI - [Validation of geriatric depression scale in a general outpatient clinic]. AB - OBJECTIVE: The Geriatric Depression Scale for screening depressive symptoms in the elderly has not been assessed in elderly outpatients who seek primary health care in Brazil. The objective was to determine the validity of the Short Scale for Major Depressive Episode or Dysthymia (GDS-15) in elderly outpatients. METHODS: The scale was applied in 302 subjects with 65 years and older and then examined by an independent geriatrician, blinded to the results. Major depression and dysthymia were diagnosed using the Diagnostic and Statistical Manual of Mental Disorders-IV criteria. Sensitivity and specificity were calculated at several cutoff values and a Receiver Operating Characteristic curve was plotted. RESULTS: The best equilibrium was at the cutoff value of 5/6 showing 81% sensitivity and 71% specificity; the area under the Receiver Operating Characteristic curve was 0.85 (95% CI: 0.79-0.91). CONCLUSIONS: The GDS-15 can be used for screening depressive symptoms in Brazilian elderly outpatients. The previously suggested cutoff value of 5/6 is adequate. PMID- 16341402 TI - [Drug use by the elderly in Southern Brazil]. AB - OBJECTIVE: To describe self-reported drug use by the elderly, evaluating the occurrence of polypharmacy and the influence of sociodemographic and health conditions on the use of medications. METHODS: A cross-sectional study comprising a sample of 215 elderly was carried out in Porto Alegre, Southern Brazil, in 2001 and 2002. Data on the use of medications were collected using questionnaires filled out during home visits. Drugs were classified according to the Anatomical Therapeutical-Chemical Classification System. RESULTS: Of all respondents, 141 (66%) were women, 117 (54%) aged between 60 and 70 years old, 157 (73%) were white, 115 (53%) lived with a partner, and 145 (67%) had some schooling. The prevalence of the use of medications by the elderly was 91% (n=195). In the week previous to the interview, 697 drugs were used, an average of 3.2 drugs per person (SD=2.5). A total of 187 (87%) subjects had attended at least one medical visit in the last year and 71 (33%) confirmed the use of over-the-counter drugs. Polypharmacy was evidenced in 57 (27%) cases. CONCLUSIONS: There is a pattern of increased drug use by the elderly aged 60 years or more who lives in the study community and slight differences were determined by their health conditions and socioeconomic factors. PMID- 16341403 TI - [Association between primary dental care and municipal socioeconomic indicators]. AB - OBJECTIVE: To test associations between primary dental care indicators and municipal socioeconomic and dental service provision indicators. METHODS: An ecological study was carried out in the 293 municipalities of the State of Santa Catarina, Brazil, between 2000 and 2003. The primary dental care indicators utilized were: (1) coverage; (2) the ratio between preventive dental procedures and the size of the population aged zero to 14 years; and (3) the ratio between the numbers of extractions of permanent teeth and individual dental procedures within primary dental care. The variables investigated were: number of dentists per 1,000 inhabitants; number of dentists within the public health service per 1,000 inhabitants; fluoridation of the water supply; child development index; human development index; and the size of the population in the municipality. Analyses were performed using the Kruskall-Wallis and Chi-square tests. The Spearman test was used to evaluate correlations between the variables. RESULTS: The coverage was 21.8%, the ratio of preventive dental procedures among the population aged zero to 14 years was 0.37 and the proportion of tooth extractions in relation to the total number of individual dental procedures was 11.9 %. Lower rates of tooth extraction were associated with higher numbers of dentists within the public health service (p<0.01). Higher rates of tooth extraction were associated with lower human development indices for the municipalities (p<0.01). CONCLUSIONS: Greater coverage was associated with higher numbers of dentists within the public health system. Municipalities with worse socioeconomic conditions were associated with greater tooth extraction rates. Oral health policies must target municipalities that present worse socioeconomic indicators. PMID- 16341404 TI - [Prevalence of trachoma in preschool and schoolchildren in the city of Sao Paulo]. AB - OBJECTIVE: To assess the prevalence of trachoma among preschool and school children of public schools to give new focus to control programs. METHODS: An epidemiological survey was carried out in Sao Paulo City in 1999. Children between four and 14 years old were selected by a cluster sampling where school shift was the sampling unit. External eye examination was conducted to detect trachoma. RESULTS: A total of 27,091 children were examined and 597 cases of trachoma were found (2.2%; 95% CI: 1.86-2.55). The prevalence ranged from 0.4% to 4.2% in 10 city areas. The trachoma detection rate in the household contacts examined was 8.7%. Follicular trachoma was found in 99% of the cases and intense trachoma in 1.0%. It was observed that 21.8% of the cases were asymptomatic. CONCLUSIONS: Though the trachoma prevalence was low, the occurrence of severe cases points out to the likelihood of cicatricial trachoma cases in the future if they are not adequately treated and controlled. The great difference in the prevalences in different city areas indicates the need for strengthening epidemiological surveillance activities. PMID- 16341405 TI - [Epidemiology of sickle cell disease hospital admissions in Brazil]. AB - OBJECTIVE: Sickle cell disease is a hereditary disease, which affects mainly the black population. The aim of the present study was to analyze hospital admissions due to acute events resulting from sickle cell disease, at the epidemiological and clinical levels. METHODS: The study population included 9,349 patients with sickle cell disease admitted to hospitals in Bahia, Rio de Janeiro, and Sao Paulo, between 2000 and 2002. The national hospital database of the Brazilian Healthcare System was used. Response variables were death and mean duration of hospital admission. Covariables included sex, age, type of admission, and hospital legal status. Proportions were compared using Chi-square or Fischer tests; for continuous variables, the Mann-Whitney or Kruskall-Wallis tests were used. RESULTS: Median age ranged between 11.0 and 12.0 years, and 70% of admissions were of patients below age 20 years. Length of stay varied with age and type of admission. Emergency rooms were the most frequent form of admission (65.6 to 90.8%). Mortality was higher among adults. Median age of death was low (26.5 to 31.5 years). CONCLUSIONS: Our results confirm the high morbidity among youngsters and show a predominance of death among young adults. PMID- 16341406 TI - HIV infection and AIDS in a small municipality in Southeast Brazil. AB - OBJECTIVE: Studies on the aspects of HIV infection in small Brazilian municipalities are invaluable to appropriately design control strategies, better allocate resources, and improve health care services. The objective of the study was to assess the clinical and epidemiological aspects of HIV infection in a small municipality. METHODS: A descriptive study was carried out in Miracema, a small municipality in the northwestern area of the state of Rio de Janeiro, Brazil, between July 1999 and December 2003. All HIV-infected adult patients followed up at the local HIV/AIDS Program were included. Clinical and epidemiologic characteristics were prospectively assessed through standardized questionnaires. RESULTS: A total of 65 adult patients who attended the local HIV/AIDS Program were analyzed. Most (34) were women (male to female ratio: 0.9). An absolute predominance of patients who were born in Miracema or neighboring municipalities (94%), lived in Miracema (90.7%), were single (70.8%), attributed the acquisition of HIV infection to unprotected heterosexual intercourse (72.3%) and had a past history of snorting cocaine (27.7) was found Central nervous system disorders (including five cases of cryptococcal meningitis) and acute pulmonary pneumocystosis-like respiratory failure were major causes of morbidity. Most patients (56.9%) were at presented in advanced stages of HIV infection. CONCLUSIONS: The predominance of patients on advanced stages of HIV infection suggest the existence of a large pool of undiagnosed cases in the community. A major feature of the cohort was an inverted male to female ratio. Further investigations over a broader geographic area are urgently needed for better understanding the clinical and epidemiological characteristics of HIV infection in small Brazilian municipalities and rural areas. PMID- 16341407 TI - [Internalized oppression and high-risk sexual practices among homosexual and bisexual males, Mexico]. AB - OBJECTIVE: To assess the relationship between engaging in high-risk sexual practices for HIV infection and the internalized oppression of homo and bisexual males due to the prejudice and discrimination they face. METHODS: An observational cross-sectional study was carried out between September and November 2001. A questionnaire was applied to 318 males who sought services and institutions in Mexico City. The studied variables were: internalized homophobia, perception of stigma, concealing sexual orientation, and sexual practices in the six months prior to the survey. The relationship (odds ratios) between forms of internalized oppression with high-risk sexual practices was estimated using logistic regression models adjusted for confounders. RESULTS: Of all respondents, 256 reported having had at least one sexual relationship in the last six months, 50 reported having none, and 12 did not answer this question. More than 30% had not used condoms when they engaged in insertive or receptive anal sex. Internalized homophobia was associated with higher risk of drug consumption before or during sexual encounters, unprotected insertive/receptive anal sex, and swallowing semen from other men. CONCLUSIONS: Engaging in high-risk sexual practices is still frequent among homo and bisexual males. Of the three forms of internalized oppression studied, homophobia was associated with higher high-risk sexual practices. This information may be incorporated into the design of programs for promoting condom use among homo and bisexual males. PMID- 16341408 TI - [Relationship between low birthweight and air pollution in the city of Sao Paulo, Brazil]. AB - OBJECTIVE: Air pollution has been investigated as a potential determinant for low birthweight. The aim of the present study was to study the effect of air pollution on birthweight. METHODS: We analyzed all deliveries by mothers living in the municipality of Sao Paulo, Southeastern Brazil, between 1998 and 2000. We estimated the prevalence of low birthweight according to newborn, mother, and delivery characteristics. Only births occurring in the most central districts of the city were analyzed, totaling 311.735 events. For the evaluation of the effects of air pollution, we excluded preterm and multiple deliveries. Pollutants analyzed were ozone (O3), sulfur dioxide (SO2), nitrogen dioxide (NO2), suspended particles (PM10), and carbon monoxide (CO). The effect of maternal exposure to air pollution on birthweight was evaluated using linear and logistic regression. RESULTS: A total of 4.6% of newborns weighed less than 2,500 g at birth. Maternal exposure to CO, PM10, and NO2 during the first trimester of pregnancy was significantly associated with decreased birthweight. CONCLUSIONS: Our results reinforce the notion that maternal exposure to air pollution during the first trimester of pregnancy may contribute to lesser weight gain in the fetus. PMID- 16341409 TI - Pesticides and respiratory symptoms among farmers. AB - OBJECTIVE: Despite the intensive use of pesticides in agriculture there are few studies assessing the risk of respiratory conditions from this exposure. The study aimed at quantifying the prevalence of respiratory symptoms among farmers and evaluating its relationship with occupational use of pesticides and the prevalence of respiratory symptoms. METHODS: A cross-sectional study was conducted among 1,379 farmers from two municipalities of Southern Brazil in 1996. Frequency and type of chemical exposure and pesticide poisoning were recorded for both sexes. All subjects aged 15 years or older with at least 15 weekly hours of agricultural activity were interviewed. An adapted questionnaire developed by the American Thoracic Society was used for the assessment of respiratory symptoms. Multivariate logistic regression analysis was carried out. RESULTS: More than half (55%) of interviewees were male. The prevalence of asthma symptoms was 12% and chronic respiratory disease symptoms was 22%. Higher odds ratios for both asthma (OR=1.51; 95% CI: 1.07-2.14) and chronic respiratory disease (OR=1.34; 95% CI 1.00-1.81) symptoms were found in women. Logistic regression analysis identified associations between many forms of exposure to pesticides and increased respiratory symptoms. Occurrence of pesticide poisoning was associated with higher prevalence of asthma symptoms (OR=1.54; 95% CI: 1.04-2.58) and chronic respiratory disease symptoms (OR=1.57; 95% CI: 1.08-2.28). CONCLUSIONS: In spite of causality limitations, the study results provide evidence that farming exposure to pesticides is associated with higher prevalence of respiratory symptoms, especially when the exposure is above two days per month. PMID- 16341410 TI - [Sensitivity, specificity and predictive value of hearing complaints]. AB - This study assessed the sensitivity, specificity and predictive value of hearing complaints in outpatients attending an university ear, nose and throat clinic. A cross-sectional study was carried out in Canoas, Southern Brazil, from 1999 to 2002. Seven-hundred and ninety-five subjects who underwent the audiometric test were analyzed. The results showed that the complaint of hearing loss presented 80.9% sensitivity, 69.6% specificity and 86.5% positive predictive value and 60.4% negative predictive value. Despite its subjective character, the audiometric test can be considered a reliable diagnostic tool and should be used due to its low cost compared to the new technologies. PMID- 16341411 TI - [Dengue: actions to fight vectors]. PMID- 16341412 TI - [Death causes according to skin color and gender in the state of Sao Paulo]. PMID- 16341413 TI - [Biosecurity]. PMID- 16341414 TI - [Food quality: program catches businessmen's attention and advises handlers]. PMID- 16341415 TI - Diet and feeding of fish from Grande River, located below the Volta Grande Reservoir, MG-SP. AB - We compare the classic model of feeding of tropical fish by means of six bimonthly samplings using gillnets of varying mesh sizes that were inspected every twelve hours throughout a forty-eight hour period. The stomachs of the fish caught were classified in three categories according to quantity of food found. The amount of fat in the visceral cavity with respect to the energetic reserve deposition was also studied. The relative frequencies of the different categories of stomach repletion and fat deposition were examined for patterns of feeding seasonality. The stomachs considered full were examined to record diet composition. To assess the relative importance of the different food resources, we applied Feeding Importance Degree (FID), which is a useful index when difficulties exist in determining a common basis for volume, number, or weight of a given food item in different species, a common problem when dealing with fish species having different feeding habits. The fish species whose stomach contents were analyzed using the FID index were Serrasalmus spilopleura (Characidae), L. prolixa (Loricaridae), Schizodon nasutus (Anostomidae), and Pimelodus maculatus (Pimelodidae). Our findings indicate some contrasting elements, in dietary composition in relation to the classic model for tropical rivers. These factors include the importance of aquatic macrophytes, the lack of piscivorous species, and a lesser presence of allochthonous vegetation in the diet of the species studied. PMID- 16341416 TI - Reproductive seasonality of fishes from a lotic stretch of the Grande River, high Parana river basin, Brazil. AB - We searched for correlations between water level, rainfall, temperature, and reproductive activity of fishes in a lotic stretch of the Grande River below the Volta Grande Dam. Gillnets were used to catch the fish on six field journeys made bimonthly between April 1998 and February 1999. Fish gonads were classified according to maturation grade and the amount of mature or spent gonads was correlated with rainfall and air temperature. Most fish species showed seasonal reproduction (between October and February). The relative frequency of fully mature and spent gonads significantly correlated with rainfall (Spearman r = 0.94, p = 0.005) and air temperature (r = 0.84, p = 0.036), showing the importance of these factors in regulating the reproductive cycle of fishes in this location. Nevertheless, dam operation resulted in minimal water level variation that did not significantly correlate with reproductive activity (r = 0.43, p = 0.396). PMID- 16341417 TI - Small mammals of Chapada dos Veadeiros National Park (Cerrado of Central Brazil): ecologic, karyologic, and taxonomic considerations. AB - This work is based on a survey of small mammals carried out in the Chapada dos Veadeiros National Park, a natural reserve located in the mountains of the Planalto Central Goiano in the Cerrado of Central Brazil. The 227 specimens collected represented six marsupial and 13 rodent species. Taxonomic, karyologic, and ecologic considerations are present and discussed in the present work. Our data reflected the faunal heterogeneity with respect to both elevation and vegetation because only eight of the 19 species were collected at both high and low elevations. The composition of the small mammal fauna of the park is influenced by predominance of forest formations at low elevations and cerrado with rupestrian areas at high elevations. Presence of endemic species and one undescribed demonstrated that the cerrado has an endemic fauna and a little known diversity of small mammals. PMID- 16341418 TI - Fruit consumption and seed dispersal of Dimorphandra mollis Benth. (Leguminosae) by the lowland tapir in the cerrado of Central Brazil. AB - Fruit phenology observations and consumption of Dimorphandra mollis (Leguminosae) were analyzed during seven months in an area of cerrado stricto sensu. We analysed 81 fecal samples collected at six different places of lowland tapirs (Tapirus terrestris) in central Brazilian cerrado. In addition, from the feces of five tapirs at the Brasilia Zoo to which fruit had been offered, seeds were collected and used in germination tests. The results suggest that the tapir is an important fruit consumer and a potential seed disperser of D. mollis. In the field, however, fruit consumption was found to be very low, probably because of both fruit palatability and the low density of frugivores, especially tapirs. The possibility that the original dispersal agents of D. mollis seeds belonged to the South American Pleistocene megafauna is discussed. PMID- 16341419 TI - Effect of temperature on the development and viability of Gryon gallardoi (Brethes) (Hymenoptera: Scelionidae) parasitizing Spartocera dentiventris (Berg) (Hemiptera: Coreidae) eggs. AB - The development and viability of Gryon gallardoi (Brethes) (Hym.: Scelionidae) in Spartocera dentiventris (Berg) (Hem.: Coreidae) eggs were studied under four temperatures: 15, 20, 25, and 30 +/- 1 degree C, with a 12-h photophase. No parasitoid developed at 15 degrees C. Otherwise, viability reached 98.8% without varying significantly over the temperature range tested. The duration of development for males and females was inversely proportional to the temperature increase, varying respectively from 46.2 +/- 0.13 and 47.1 +/- 0.11 days (20 degrees C) to 13.3 +/- 0.07 and 13.4 +/- 0.06 days (30 degrees C). Males developed faster than females. The values estimated for the lowest thermic thresholds of development and the thermic constants were 15.5 degrees C and 185.19 DD for males and 15.6 degrees C and 192.31 DD for females, respectively. Given the average weather conditions in Porto Alegre, RS (30 degrees 01' S and 51 degrees 13' W), Brazil, G. gallardoi could annually produce 8.54 and 8.07 generations of males and females, respectively. The low rates of parasitism observed in the field during the first generation of its host may be due to the small number of G. gallardoi generations in this period. PMID- 16341420 TI - Effects of fragmentation on Thamnophilus stictocephalus (Aves, Thamnophilidae) in semideciduous forest of Alter-do-Chao, Para. AB - Effects of fragmentation on biodiversity have received much attention in recent decades, as fragmentation can greatly reduce viable areas for living organisms. We studied its effect on Thamnophilus stictocephalus (Thamnophilidae), an understory bird, in semideciduous forest fragments in Alter-do-Chao, Santarem, Para. We tested whether the density of Thamnophilus stictocephalus was a function of fragment size and shape, density of vegetation, or arthropod biomass. Density of Thamnophilus was positively related to fragment size, but not to the other factors analyzed. Arthropod biomass was positively related to fragment size. The density of T. stictocephalus in fragments was significantly higher than it was in continuous forest. Fragmentation processes had a pronounced effect on the relative density of T. stictocephalus. PMID- 16341421 TI - Ant and termite mound coinhabitants in the wetlands of Santo Antonio da Patrulha, Rio Grande do Sul, Brazil. AB - This paper reports on ant and termite species inhabiting the mounds (murundus) found in three wetland sites in Santo Antonio da Patrulha. Ants and termites were found in 100% of the mounds of two sites and in 20% of those in the third site. Colonies of Camponotus fastigatus were found inhabiting all the mounds, while colonies of Brachymyrmex sp., Linepithema sp., Pheidole sp., and/or Solenopsis sp. were collected in less than 30% of the mounds. In the mounds of the three sites, colonies of Anoplotermes sp. and/or Aparatermes sp. termites were found together with the ant colonies. Another cohabiting termite species, Cortaritermes sp., was found only in the mounds of one site. The results suggest that C. fastigatus is the species building the mounds, with the other species, whether ants or termites, being the inquilines. PMID- 16341422 TI - Spatial synchrony of a highly endemic fish assemblage (Segredo Reservoir, Iguacu River, Parana State, Brazil). AB - In this study, patterns of spatial synchrony in population fluctuations (cross correlation) of an endemic fish assemblage of a Neotropical reservoir (Segredo Reservoir, Iguacu River, Parana State, Brazil) were reported. First, the level of population synchrony for 20 species was estimated. Second, population synchrony was correlated, using the Mantel test, with geographical distances among sites (n = 11) and also environmental synchrony (temperature). Nine species presented significant correlations between spatial synchrony and geographic distances (Astyanax sp. b, Astyanax sp. c, Pimelodus sp., Hoplias malabaricus, Crenicichla iguassuensis, Hypostomus derbyi, Hypostomus myersi, Rhamdia branneri, and R. voulezi). Considering the ecology of the species and the significant relationship between population and environmental synchronies, it seems that environmental stochasticity is the most plausible hypothesis in explaining the observed synchrony patterns. PMID- 16341423 TI - Flood pulse influence and anthropic impact on the chemical composition and energy content of Oryza glumaepatula in an Amazonian lake. AB - The aim of this research was to study the flood pulse influence and the anthropic impact caused by bauxite tailings on the chemical composition of O. glumaepatula in Batata lake (PA, Brazil). Sampling was carried out in stands of O. glumaepatula in the low-water, filling, high-water, and drawdown periods in impacted and natural areas of Batata lake. During the low-water and drawdown periods the stands of O. glumaepatula were exposed, and in the filling and high water periods the water depth was respectively 1.4 and 3.8 m. The collected material was dried at 70 degrees C, ground, and concentrations of total phosphorus, total nitrogen, organic carbon, and energy content were determined. The results indicate that the biomass increase, caused by the rise in water level, has a dilution effect on nitrogen and phosphorus concentrations in O. glumaepatula. The energy contents did not present significant differences in any of the studied periods. The results suggest that from the low water to filling period, nitrogen becomes more limiting to O. glumaepatula in the impacted area, whereas phosphorus becomes more limiting in the natural area. The population of O. glumaepatula contributes to the recovery of the impacted area of Batata lake as the detritus from this species accumulates over the sediment. This accumulation impedes future re-suspension of the bauxite tailings and increases the organic matter and nutrient concentrations in the impacted sediment. PMID- 16341424 TI - The annual reproductive cycle of the snail Megalobulimus abbreviatus (Bequaert, 1948) (Gastropoda, Pulmonata). AB - Morphological changes in the sexual organs of the pulmonates were observed throughout a year and correlated with reproductive-cycle periods. Reproductive organ weights of the snail Megalobulimus abbreviatus were recorded seasonally and gonad sections were analyzed morphologically. The weights were used to obtain the organosomatic index. Mean oocytic diameter and oocytic maturation index were based on gonad sections. It was concluded that M. abbreviatus is an iteroparous snail whose annual reproductive cycle is characterized by mating and egg laying throughout spring and early summer, and also by reproductive system preparation, occurring over the remainder of the summer until the end of winter, for a new breeding season. PMID- 16341425 TI - Colony strength and queen replacement in Melipona marginata (Apidae: Meliponini). AB - Physogastric queens of Melipona marginata were removed from their colonies in order to verify the acceptance of a new queen by workers. Colony strength was evaluated according to queen oviposition rate and comb diameters. Replacement was observed seven times. Its occurrence and speed related positively to colony strength, independently of queen's age. In weak colonies, queen replacement was observed only once, following colony population increase that occurred after introduction of combs from another colony. Worker oviposition after queen removal was observed three times: in a strong colony with virgin queens and males, and in two of the weak colonies. In the first two or three days of new queen oviposition, during which most of the eggs were eaten by the queen, worker oviposition preceded almost all provisioning and oviposition processes (POPs). After this period, worker oviposition decreased until it reached around 25% of the POPs. Daily oviposition rate of young queens decreased or was even interrupted by hatching of their first brood. PMID- 16341426 TI - Eye pigments of the blood-sucking insect, Triatoma infestans Klug (Hemiptera, Reduviidae). AB - The pigmentation of black (wild) and red (mutant) eyes of Triatoma infestans was studied spectrophotometrically and compared with red-eyed (wild) and white-eyed (mutant) forms of Drosophila melanogaster. The spectral absorption profiles of the black and red eye pigments of T. infestans were similar to each other and to that of the wild-type eyes of D. melanogaster. The similarity to the wild form of D. melanogaster indicated that both eye forms of T. infestans contained ommochromes of the xanthommatin type, a finding confirmed by ascending paper chromatography. Pteridines, melanins, and ommins were not detected as eye pigments in T. infestans. The eye color difference in T. infestans was assumed to be a function of the xanthommatin concentration, with a smaller content of ommochrome in red eyes, although this probably did not affect the insect's visual acuity. These data support other findings regarding the similarities between black- and red-eyed specimens of T. infestans for other characteristics. PMID- 16341427 TI - Anatomy and ontogeny of Pterodon emarginatus (Fabaceae: Faboideae) seed. AB - The aim of this study was to describe the anatomy and ontogeny of Pterodon emarginatus seed using the usual techniques. The ovules are campilotropous, crassinucelate, and bitegmic. The following processes occur during integument development: anticlinal divisions and phenolic compound accumulations in the exotesta, whose cells become palisade; predominantly periclinal divisions and cell expansion in the mesotesta, where the rapheal bundle differentiates; differentiation of the hourglass-cell layer adjacent to the palisade; fusion of outer and inner integuments, which remain individualized structures only at the micropylar end; and intense pectin impregnation in the mesotesta thicker walls with lignification restricted to the xylem. At the hilar pole, the Faboideae seed characteristic structure develops, with double palisade layer, subhilar parenchyma, and tracheid bar. The younger nucellus shows thicker pectic cell walls and is consumed during seed formation. The endosperm is nuclear and, after cellularization, shows peripheral cells with dense lipid content; the seeds are albuminous. The axial embryo shows fleshy cotyledons, which accumulate lipid and protein reserves; starch is rare. Although the seed structure is characteristic of the Fabaceae, the inner integument coalesces into the outer integument without being reabsorbed. PMID- 16341428 TI - Relation between small-mammal species composition and anthropic variables in the Brazilian Atlantic Forest. AB - Anthropic activities are frequently related in many ways to forest fragmentation and alteration of natural communities. In this study, we correlate the presence of hunting, tourism activity, agriculture/pasturing, and the distance of the study sites to the nearest human residences with the species composition of small Atlantic forest mammals. To do this, we utilize a multiple regression analysis of similarity matrices. The presence of both agriculture/pasturing and human residences near the study sites proved to be determinant factors in species composition of small mammals of the studied areas. Working with socioeconomic variables related directly with the study site could be a reliable and a direct way to predict the influence of human presence and entailed activity on small mammal communities. PMID- 16341429 TI - Distribution and density of bacteria in subtropical flooded rice growing areas in Brazil. AB - Since microorganisms are important components in the structure and function of water systems, the present paper reports work done to identify groups making up bacterial communities present in rice irrigation water. To this end, water samples were collected in five rice-growing areas of Rio Grande do Sul State, Brazil. The identification of the main bacterial groups found in flooded areas was carried out based on colony morphology, taking into account shape, elevation, and edges. Internal and external cell structure was characterized as cocci or rod, gram-positive or gram-negative, and spore forming or not. The results obtained regarding density and bacterial distribution in the water samples from rice growing areas of RS showed a higher abundance of morphotypes of bacterial colonies in the irrigation channels (F(1,12) = 8.74; p < 0.05) when compared to the culture plots in the five production regions, and in the final culture phase (F(1,12) = 8.86; p < 0.05). In the five rice production areas the occurrence of gram-positive and spore-forming rods was prevalent. Rods showed more abundance (F(1,26) = 15.12; p < 0.05) in the early culture phase, with a significant prevalence of gram-positive rods (F(1,26) = 25.99; p < 0.05) in all rice growing regions under investigation. The group of spore-forming bacteria was significantly more abundant (F(1,30) = 14.06; p < 0.05) when compared with the non-spore-forming ones in all investigated regions. Data showed that the irrigated rice crop affects the density and morphotype colonies in the bacteria found in the water used to flood rice-growing plots. PMID- 16341430 TI - Parasitism of Cichlasoma paranaense Kullander, 1983 and Gymnotus carapo Linnaeus, 1814 by Clinostomum complanatum (Rudolphi, 1814) metacercariae in the Taquari river. AB - The infection, known as yellow spot disease, produced by metacercariae of Clinostomum spp. was studied in fishes of the Taquari river, located in Jataizinho, Parana State, Brazil. A total of 1,582 specimens, belonging to 36 species, were collected between March 1999 and April 2001. Yellow spot disease was observed only in Gymnotus carapo Linnaeus, 1814 (Gymnotiformes, Gymnotidae) and Cichlasoma paranaense Kullander, 1983 (Perciformes, Cichlidae). This parasitism was generated by metacercariae of Clinostomum complanatum (Rudolphi, 1814) (Digenea, Clinostomidae). Among the 88 specimens of G. carapo examined, 7 (prevalence = 8%) had cysts of the parasite. Four of them were captured in July 1999 and three in October 1999. In the other months, no specimen found was infected. Using relative condition factor (Kn) analysis, it was determined that both infected and non-infected specimens had a total weight equal to the theoretically expected value for each total length (Kn = 1.0). Among 56 individuals of Cichlasoma paranaense, 6 (prevalence = 10.7%) had between 1 and 27 metacercariae of C. complanatum (mean intensity of infection = 9.3 +/- 9.6). In March 1999 and April of both 2000 and 2001, the specimens examined were not infected. The infected fish had a total weight higher than the expected value (Kn > 1.0), while the non-infected fish had a weight equal to the expected value (Kn = 1.0). PMID- 16341431 TI - Taxonomy, ecology, and geographical distribution of the species of the genus Thermocyclops Kiefer, 1927 (Copepoda, Cyclopoida) in Sao Paulo State, Brazil, with description of a new species. AB - The taxonomy and ecology of the cyclopoid copepod genus Thermocyclops were studied. Samples were collected in 207 water bodies located in the 22 hydrographic basins of Sao Paulo State, Brazil, including large reservoirs, small and shallow lakes, and ponds and rivers. The genus Thermocyclops inhabits mainly water bodies within a limnetic region. Four species were found, of which one is new: Thermocyclops iguapensis, which occurred in the reservoirs of the Ribeira do Iguape and Paraiba do Sul basins. The description of the new species and the geographical distribution of all four species in Sao Paulo State are presented. Thermocyclops decipiens was the most frequent species, occurring in 71% of the water bodies within a limnetic region. This species is characteristic of eutrophic environments where it can occur in great abundance, whereas Thermocyclops minutus is characteristic in oligotrophic systems. Thermocyclops inversus and Thermocyclops iguapensis n. sp. were not common but can occur together with Thermocyclops decipiens. PMID- 16341432 TI - Community ecology of metazoan parasites of the anchovy Anchoa tricolor (Osteichthyes: Engraulidae) from the coastal zone of the State of Rio de Janeiro, Brazil. AB - Between October 2001 and March 2002, 103 specimens of A. tricolor from Angra dos Reis (23 degrees 01' S, 44 degrees 19' W), in the coastal zone of the State of Rio de Janeiro, Brazil, were analyzed in order to study their metazoan parasite infracommunities. Ten species of metazoan parasites were collected: 4 digeneans, 1 cestode, 1 acantocephalan, 2 nematodes, 1 copepod, and 1 hirudinean; 77.7% of the fishes were parasitized by one or more metazoan, with a mean of 3.5 +/- 6.2 parasite/fish. Digenean was the most dominant with 4 species that accounted for 53.2% of the total parasites collected; Ergasilus sp. was the most abundant species. Abundance and prevalence of Parahemiurus merus (Linton, 1910) were positively correlated with the total length of host. Relationships between total body length of fish and both total parasite abundance and mean parasite species richness were observed. Mean parasite diversity of species was correlated to host's total length, with significant differences found between male and female fishes. Two pairs of larval species showed significant positive association and covariation. The metazoan parasite infracommunities of A. tricolor presented dominance of larval endoparasites; correlation of parasite abundance, diversity, and species richness with host total length; and low number of parasite interspecific relationships. The parasite community of A. tricolor showed some similarities with the parasite community of another South American Atlantic engraulid. PMID- 16341433 TI - Genetics of Euglossini bees (Hymenoptera) in fragments of the Atlantic Forest in the region of Vicosa, MG. AB - With uncontrolled deforestation, forest fragments remain, which in most cases are in different stages of regeneration and present isolated populations. In the present study we analyzed the genetic patterns of Eulaema nigrita populations in seven Atlantic Forest fragments of different sizes and successional stages in the region of Vicosa, MG. This was done by RAPD molecular markers. We observed that the area of the fragments had no effect on the genetic variability of E. nigrita in the direction predicted by meta-population models. Medium-sized well-preserved woods presented the lowest variability, whereas large and small woods were statistically identical. The evidence supports the notion that rural areas present greater dispersal among fragments, implying greater similarity between the populations of fragments located in rural areas when compared to fragments in urban areas. PMID- 16341434 TI - Seed cleaning of Strychnos ramentifera (Loganiaceae) by ants in Maraca Island, Brazilian Amazonia. PMID- 16341435 TI - Determination of Duffy phenotype of red blood cells in Dasypus novemcinctus and Cabassous sp. PMID- 16341436 TI - General optimal euclidean Sobolev and Gagliardo-Nirenberg inequalities. AB - We prove general optimal euclidean Sobolev and Gagliardo-Nirenberg inequalities by using mass transportation and convex analysis results. Explicit extremals and the computation of some optimal constants are also provided. In particular we extend the optimal Gagliardo-Nirenberg inequality proved by Del Pino and Dolbeault 2003 and the optimal inequalities proved by Cordero-Erausquin et al. 2004. PMID- 16341437 TI - Complementary Lagrangians in infinite dimensional symplectic Hilbert spaces. AB - We prove that any countable family of Lagrangian subspaces of a symplectic Hilbert space admits a common complementary Lagrangian. The proof of this puzzling result, which is not totally elementary also in the finite dimensional case, is obtained as an application of the spectral theorem for unbounded self adjoint operators. PMID- 16341438 TI - On Gymnodactylus amarali Barbour, 1925, with the description of a new species (Sauria, Gekkonidae). AB - Gymnodactylus amarali Barbour, 1925, was previously considered to be a subspecies of G. geckoides, with a wide distribution in the Brasilian cerrados. Examination of a specimen from Alto Parnaiba, Maranhao, near the type locality (Engenheiro Dodt, Piaui), indicates that it is a proper species, apparently limited to the upper Parnaiba basin. The form previously identified as G. geckoides amarali is described as a new species, G. carvalhoi, type locality Ipueiras, State of Tocantins, thus diagnosed: color pattern plain or, more often, with moderately contrasted ocelli; dorsal tubercles in 13-16 poorly organized longitudinal rows (mode 14, 72%); 31-49 tubercles in a paramedian row; 17-22 transverse rows of ventral scales; 13-18 infradigital lamellae on toe IV; tail longest in the genus. The new species is statistically compared to parapatric G. geckoides, widespread in the caatingas. Although only one meristic character (number of tubercle rows) is by itself diagnostic, the species are easily told apart. It is thought on provisional evidence that they are better considered for the time being as full species, not subspecies. A brief consideration is made of the speciation model that seems suitable, to wit, parapatric. PMID- 16341439 TI - Constraints to the implementation of effective environmental management in coastal areas of developing countries. AB - This paper addresses the limitations the scarcity of reliable scientific information poses to the implementation of effective and sustainable coastal management programmes in developing countries. Alternatives to the current monitoring approaches are suggested as to improve information level on the state of the environment and to decrease data gap on past conditions. The paper aims at encouraging the redesign of monitoring practices in developing countries as to be ground on the best actual scientific knowledge. PMID- 16341441 TI - Evolutionary change--patterns and processes. AB - The present review considered: (a) the factors that conditioned the early transition from non-life to life; (b) genome structure and complexity in prokaryotes, eukaryotes, and organelles; (c) comparative human chromosome genomics; and (d) the Brazilian contribution to some of these studies. Understanding the dialectical conflict between freedom and organization is fundamental to give meaning to the patterns and processes of organic evolution. PMID- 16341442 TI - The specificity of interactions between proteins and sulfated polysaccharides. AB - Sulfated polysaccharides are capable of binding with proteins at several levels of specificity. As highly acidic macromolecules, they can bind non-specifically to any basic patch on a protein surface at low ionic strength, and such interactions are not likely to be physiologically significant. On the other hand, several systems have been identified in which very specific substructures of sulfated polysaccharides confer high affinity for particular proteins; the best known example of this is the pentasaccharide in heparin with high affinity for antithrombin, but other examples may be taken from the study of marine invertebrates: the importance of the fine structure of dermatan sulfate (DS) to its interaction with heparin cofactor II (HCII), and the involvement of sea urchin egg-jelly fucans in species specific fertilization. A third, intermediate, kind of specific interaction is described for the cell-surface glycosaminoglycan heparan sulfate (HS), in which patterns of sulfate substitution can show differential affinities for cytokines, growth factors, and morphogens at cell surfaces and in the intracellular matrix. This complex interplay of proteins and glycans is capable of influencing the diffusion of such proteins through tissue, as well as modulating cellular responses to them. PMID- 16341443 TI - Haematophagous arthropod saliva and host defense system: a tale of tear and blood. AB - The saliva from blood-feeding arthropod vectors is enriched with molecules that display diverse functions that mediate a successful blood meal. They function not only as weapons against host's haemostatic, inflammatory and immune responses but also as important tools to pathogen establishment. Parasites, virus and bacteria taking advantage of vectors' armament have adapted to facilitate their entry in the host. Today, many salivary molecules have been identified and characterized as new targets to the development of future vaccines. Here we focus on current information on vector's saliva and the molecules responsible to modify host's hemostasis and immune response, also regarding their role in disease transmission. PMID- 16341445 TI - Geochemical modeling of gold precipitation conditions in the Bloco do Butia Mine, Lavras do Sul/Brazil. AB - A geochemical modeling of gold deposition was performed using the EQ3/EQ6 software package using conditions inferred from geological, petrographic, geochemical and fluid inclusion data from the Bloco do Butia gold mine, Lavras do Sul, RS. Gold in the mine occurs only in the pyrite structure (invisible gold). The pyrite occurs associated with white mica (phengite) in the zone of phyllic alteration. The process of gold deposition showed to be related to temperature and pH decrease. The pH decrease was fundamental to gold deposition by destabilization of sulfur species [Au(HS)2- , HAu(HS)2(0) and Au(HS)0] dissolved in the aqueous solution, being Au(HS)0 the main gold transporting complex. The addition of KCl is hard to accept as cause of gold precipitation because no Cl- was detected in phengite. However, the geochemical mass balance calculation resulted in the gain of some potassium in the zone of phyllic alteration. The precipitation of pyrite (+/- auriferous) may have been strongly influenced by iron availability resulting from dissolution of ferrous chlorites by the fluids responsible for phengite deposition. The low salinity in quartz grain fluid inclusions from the propylitized wall rock also indicates the little importance of chlorine as gold transporting agent. Sulfur, and not chlorine, compounds must have dominated the gold transporting complexes in the Bloco do Butia gold area. PMID- 16341444 TI - An overview of chagasic cardiomyopathy: pathogenic importance of oxidative stress. AB - There is growing evidence to suggest that chagasic myocardia are exposed to sustained oxidative stress-induced injuries that may contribute to disease progression. Pathogen invasion- and replication-mediated cellular injuries and immune-mediated cytotoxic reactions are the common source of reactive oxygen species (ROS) in infectious etiologies. However, our understanding of the source and role of oxidative stress in chagasic cardiomyopathy (CCM) remains incomplete. In this review, we discuss the evidence for increased oxidative stress in chagasic disease, with emphasis on mitochondrial abnormalities, electron transport chain dysfunction and its role in sustaining oxidative stress in myocardium. We discuss the literature reporting the consequences of sustained oxidative stress in CCM pathogenesis. PMID- 16341446 TI - Chemical and mineralogical characterization of elbaites from the Alto Quixaba pegmatite, Serido province, NE Brazil. AB - The Alto Quixaba pegmatite, Serido region, northeastern Brazil, is a 60 degrees/80 degrees SW-trending subvertical dike discordantly intruded into biotite schists of the Upper Neoproterozoic Serido Formation. It has three distinct mineralogical and textural zones, besides a replacement body that cuts the pegmatite at its central portion and in which occur, among other gem minerals, colored elbaites. Elbaites usually occur as prismatic crystals, elongate according to the c-axis, with rounded faces and striations parallel to this axis. Optically, crystals are uniaxial negative with strong pleochroism; refractive index extraordinary axis = 1.619-1.622 and ordinary axis = 1.639 1.643, birefringence between 0.019 and 0.021, average relative density of 3.07, and the following unit cell parameters: ao = 15.845 A, co = 7.085 A and V = 1540.476 A. There is alkali deficiency in the X site of 12-17%. The elbaites are relatively enriched in MnO (1.69 to 2.87%) and ZnO (up to 2.98%). PMID- 16341447 TI - On the retrieval of significant wave heights from spaceborne Synthetic Aperture Radar using the Max-Planck Institut algorithm. AB - Synthetic Aperture Radar (SAR) onboard satellites is the only source of directional wave spectra with continuous and global coverage. Millions of SAR Wave Mode (SWM) imagettes have been acquired since the launch in the early 1990's of the first European Remote Sensing Satellite ERS-1 and its successors ERS-2 and ENVISAT, which has opened up many possibilities specially for wave data assimilation purposes. The main aim of data assimilation is to improve the forecasting introducing available observations into the modeling procedures in order to minimize the differences between model estimates and measurements. However there are limitations in the retrieval of the directional spectrum from SAR images due to nonlinearities in the mapping mechanism. The Max-Planck Institut (MPI) scheme, the first proposed and most widely used algorithm to retrieve directional wave spectra from SAR images, is employed to compare significant wave heights retrieved from ERS-1 SAR against buoy measurements and against the WAM wave model. It is shown that for periods shorter than 12 seconds the WAM model performs better than the MPI, despite the fact that the model is used as first guess to the MPI method, that is the retrieval is deteriorating the first guess. For periods longer than 12 seconds, the part of the spectrum that is directly measured by SAR, the performance of the MPI scheme is at least as good as the WAM model. PMID- 16341460 TI - The retroverted uterus: ignored to date but core to prolapse. AB - The retroverted uterus has been largely ignored in urogynaecological research to date. The prevalence of the retroverted uterus is 79% more common in the urogynaecological patient population (34%) than in the general gynaecological population (19%). Its diagnosis requires the use of (a) transvaginal ultrasound with (b) an empty bladder. Recent data demonstrate that the prevalence of grade 2 4 uterine prolapse for a retroverted uterus is 4.5 times that for an anteverted uterus. Alternatively, 69% grade 2-4 uterine prolapse involves the retroverted uterus. The retroverted uterus, when diagnosed by transvaginal ultrasound (bladder empty), is far more common in urogynecology patients due to their higher incidence of prolapse. PMID- 16341461 TI - Increased expression of matrix metalloproteinase 2 in uterosacral ligaments is associated with pelvic organ prolapse. AB - The uterosacral ligaments are an important part of the pelvic support system and connective tissue alterations are thought to contribute to the development of pelvic organ prolapse (POP). The objective of this study was to compare the expression of matrix metalloproteinases (MMPs) 1 and 2 in these ligaments in women with and without POP. We analyzed the tissue samples obtained from left and/or right uterosacral ligaments of 17 women with POP and 18 controls by immunohistochemistry. There was no difference in MMP-1 expression between women with POP and those without. In contrast, the MMP-2 expression was significantly related to the presence of POP (p=0.004) rather than to age or parity. There was no difference in MMP-1 and MMP-2 expression between left and right uterosacral ligaments in women with POP compared to controls. Our findings strongly indicate that increased MMP-2 expression in uterosacral ligaments is associated with POP. PMID- 16341462 TI - A histidine kinase sensor protein gene is necessary for induction of low pH tolerance in Sinorhizobium sp. strain BL3. AB - The aim of this investigation was to identify and isolate genes involved in acid tolerance from Sinorhizobium sp. strain BL3. It was hypothesized that acid tolerance of strain BL3 could be enhanced by high level expression of certain genes involved in acid tolerance, following insertion of these genes in a multiple copy plasmid. A cosmid clone library of BL3 was introduced into BL3, and the transconjugant colonies were selected at low pH. A single cosmid containing genes for acid tolerance was isolated from 40 different colonies. By transposon insertion mutagenesis, subcloning and DNA sequencing, a gene involved in acid tolerance, actX, was identified in a 4.4-kb fragment of this cosmid. The actX mutant of BL3 showed increased acid sensitivity and was complemented by the 4.4 kb subcloned fragment. Phaseolus lathyroides seedlings inoculated with recombinant strains containing multiple copies of actX showed increased symbiotic performance at low pH. By constructing an actX::gus fusion, it was shown that actX was induced at low pH. actX encodes a putative histidine kinase sensor protein of a two-component regulatory system. The method of gene identification used in this study for isolation of actX may be applied for the isolation of other genes involved in tolerance to adverse environmental factors. PMID- 16341463 TI - Oxidation of arsenite by Thiomonas strains and characterization of Thiomonas arsenivorans sp. nov. AB - A novel bacterium, strain b6(T) (T=type strain), was isolated from a disused mine site by growth using arsenite [As(III)] as energy source in a simple mineral medium. Cells of strain b6(T) were rod-shaped, Gram-negative, non-sporulating and motile. Optimum growth occurred at temperatures between 20 and 30 degrees C, and at pH between 4.0 and 7.5. Strain b6(T) grew chemoautotrophically on As(III), sulphur and thiosulphate, and also heterotrophically on yeast extract and a variety of defined organic compounds. Several other Thiomonas strains, including the type species Thiomonas (Tm.) intermedia, were able to oxidize As(III), though only strain b6(T) and strain NO115 could grow using As(III) as sole energy source in the absence of any organic compound. The G+C content of the DNA of strain b6(T) was 65.1 mol %. Comparative small subunit (SSU) ribosomal RNA (rRNA) analysis indicated that strain b6(T) belongs to the genus Thiomonas in the beta subdivision of the Proteobacteria. It was closely related to an unnamed Thiomonas strain (NO115) isolated from a Norwegian mining site, though sequence identities between strain b6(T) and characterized Thiomonas species were less than 95%. DNA DNA hybridization between strain b6(T) and the type species of the genus Tm. intermedia showed less than 50% homology. On the basis of phylogenetic and phenotypic characteristics, strain b6(T) (DSM 16361(T), LMG 22795(T)) is proposed as the type strain of the new species Thiomonas arsenivorans, sp. nov. PMID- 16341464 TI - Cryptococcus silvicola nov. sp. from nature reserves of Russia and Portugal. AB - Nitrate-positive strains of a filobasidiaceous anamorphic yeast related to Cryptococcus cylindricus were isolated from forest litter in a Russian nature reserve and from a lichen in Portuguese one. Mycocinotyping and rDNA sequence analysis revealed that the strains represent a novel species, for which the name Cryptococcus silvicola (type strain VKM Y-2939=CBS 10099) is proposed. PMID- 16341467 TI - Primate-specific endogenous cis-antisense transcription in the human 5q31 protocadherin gene cluster. AB - Protocadherins (PCDH), localized to synaptic junctions, contribute to the formation of neuronal networks during brain development; thus, it is speculated that protocadherins may play a role in evolution of neuronal complexity. While protocadherin genes are highly conserved in vertebrates, EST evidence from the locus suggests apparently species-specific cis-antisense transcripts. Novel cis antisense transcripts, which partially overlap the PCDHalpha12 variable exon, PCDHbeta3 single-exon gene, and PCDHpsi5 unprocessed pseudogene in the human 5q31 PCDHalpha/beta/gamma gene cluster and which are coexpressed with sense-strand transcripts in fetal and adult brain, were identified computationally and validated by gene-specific strand-specific reverse transcriptase PCR (SSRTPCR) and sequencing. Absence of antisense transcripts arising from equivalent genomic locations in mouse indicates that the antisense transcripts originated in the primates after the primate-rodent divergence. Furthermore, not all expected orthologues of human sense and antisense PCDH transcripts were detected in rhesus macaque brain, implying that protocadherin expression patterns differ between primate species. RT followed by quantitative real-time PCR (QPCR) analysis of the three genes in the brain of all three species, and of the PCDHbeta15 gene paralogous to PCDHpsi5 in human and rhesus, revealed that the presence of antisense transcripts was significantly associated with lower sense expression levels across all orthologues. This inverse relationship, along with the pattern of sense and antisense coexpression in the brain, is consistent with a regulatory role for the primate-specific PCDH cis-antisense transcripts, which may represent recent evolutionary inventions modulating the activity of this conserved gene cluster. PMID- 16341468 TI - A comparative categorization of protein function encoded in bacterial or archeal genomic islands. AB - Genomes of prokaryotes harbor genomic islands (GIs), which are frequently acquired via horizontal gene transfer (HGT). Here I present an analysis of GIs with respect to gene-encoded functions. GIs were identified by statistical analysis of codon usage and clustering. Genes classified as putatively alien (pA) or putatively native (pN) were categorized according to the COG database. Among pA and pN genes, the distribution of COG functions and classes were studied for different groupings of prokaryotes. Groups were formed according to taxonomical relation or habitats. In all groups, genes related to class L (replication, recombination, and repair) were statistically significantly overrepresented in GIs. GIs of bacteria and archaea showed a distinct pattern of preferences. In archeal GIs, genes belonging to COG class M (cell wall/membrane/envelope biogenesis) or Q (secondary metabolites biosynthesis, transport, and catabolism) were more frequent. In bacterial GIs, genes of classes U (intracellular trafficking, secretion, and vesicular transport), N (cell motility), and V (defense mechanisms) were predominant. Underrepresentation was strongest for genes belonging to class J (translation, ribosomal structure, and biogenesis). Among single COG functions overrepresented in GIs were transferases and transporters. In both superkingdoms, HGT enhances genomic content by meeting demands that are independent of the studied habitats. These findings are in agreement with the complexity theory, which predicts the preferential import of operational genes. However, only specific subsets of operational genes were enriched in GIs. Modification of the cell envelope, cell motility, secretion, and protection of cellular DNA are major issues in HGT. PMID- 16341473 TI - Lumping in pharmacokinetics. AB - Pharmacokinetic (PK) models simplify biological complexity by dividing the body into interconnected compartments. The time course of the chemical's amount (or concentration) in each compartment is then expressed as a system of ordinary differential equations. The complexity of the resulting system of equations can rapidly increase if a precise description of the organism is needed. However, difficulties arise when the PK model contains more variables and parameters than comfortable for mathematical and computational treatment. To overcome such difficulties, mathematical lumping methods are new and powerful tools. Such methods aim at reducing a differential system by aggregating several variables into one. Typically, the lumped model is still a differential equation system, whose variables are interpretable in terms of variables of the original system. In practice, the reduced model is usually required to satisfy some constraints. For example, it may be necessary to keep state variables of interest for prediction unlumped. To accommodate such constraints, constrained lumping methods have are also available. After presenting the theory, we study, here, through practical examples, the potential of such methods in toxico/pharmacokinetics. As a tutorial, we first simplify a 2-compartment pharmacokinetic model by symbolic lumping. We then explore the reduction of a 6-compartment physiologically based pharmacokinetic model for 1,3-butadiene with numerical constrained lumping. The lumping methods presented here can be easily automated, and are applicable to first-order ordinary differential equation systems. PMID- 16341474 TI - Optimal blood sampling time windows for parameter estimation using a population approach: design of a phase II clinical trial. AB - The objective of this paper is to determine optimal blood sampling time windows for the estimation of pharmacokinetic (PK) parameters by a population approach within the clinical constraints. A population PK model was developed to describe a reference phase II PK dataset. Using this model and the parameter estimates, D optimal sampling times were determined by optimising the determinant of the population Fisher information matrix (PFIM) using PFIM_ _M 1.2 and the modified Fedorov exchange algorithm. Optimal sampling time windows were then determined by allowing the D-optimal windows design to result in a specified level of efficiency when compared to the fixed-times D-optimal design. The best results were obtained when K(a) and IIV on K(a) were fixed. Windows were determined using this approach assuming 90% level of efficiency and uniform sample distribution. Four optimal sampling time windows were determined as follow: at trough between 22 h and new drug administration; between 2 and 4 h after dose for all patients; and for 1/3 of the patients only 2 sampling time windows between 4 and 10 h after dose, equal to [4 h-5 h 05] and [9 h 10-10 h]. This work permitted the determination of an optimal design, with suitable sampling time windows which was then evaluated by simulations. The sampling time windows will be used to define the sampling schedule in a prospective phase II study. PMID- 16341475 TI - Ileosigmoid knotting situs inversus and splenic infarct. PMID- 16341477 TI - Open vs. closed hemorrhoidectomy. PMID- 16341478 TI - Vaginal reconstruction with vertically oriented rectus abdominus myocutaneous (VRAM) flap following APR for locally advanced rectal cancer. PMID- 16341480 TI - Serum levels of interleukin-12 in Graves' disease and their dynamic changes after surgery. AB - PURPOSE: To investigate the role of interleukin-12 (IL-12) in Graves' disease, we measured the pre- and postoperative levels of serum IL-12 in patients undergoing surgery for Graves' disease. METHODS: The subjects of this study were 73 patients with Graves' disease, admitted for surgical treatment after taking antithyroid drugs for various durations. We collected blood from 11 of these patients, 1, 3, and 6 months postoperatively, to measure the serum IL-12 levels using a Human IL 12 +p40 Immunoassay Kit. RESULTS: The preoperative levels of serum IL-12 were higher in patients with Graves' disease than in healthy controls. Based on the preoperative data, there was a significant relationship between the levels of serum IL-12 and free T3. An analysis of the postoperative time course of these 11 patients showed that the levels of serum IL-12 decreased gradually from 1 month to 6 months, postoperatively. There was also a significant correlation between the levels of serum IL-12 and soluble IL-2R, and a significant negative correlation between the levels of serum IL-12 and thyroid-stimulating hormone receptor antibody. CONCLUSION: Measurement of the levels of serum IL-12 may be a valuable immunological marker in the time course of treatment for Graves' disease. PMID- 16341479 TI - Microsatellite instability in gastrointestinal tract cancers: a brief update. AB - Microsatellite instability (MSI) was initially reported in colorectal cancer and, particularly, in hereditary nonpolyposis colorectal cancer (HNPCC). Since mutations in the genes functioning in DNA mismatch repair (MMR) were found in HNPCC kindred, this phenotype has been connected to a deficiency in MMR. The MSI(+) phenotype is associated with various human malignancies. As MSI(+) tumors appear to form a unique clinicopathological and molecular entity that is clearly distinct from that of classical colorectal tumors, which are accompanied by chromosomal instability (CIN), an exclusive pathway of tumorigenesis has been proposed in colorectal cancer. However, this scheme, comprising two mutually exclusive pathways, is now being reexamined, in light of a series of evidence accumulating in the literature, which relates to (a) distinction between high level MSI (MSI-H) and low-level MSI (MSI-L), (b) heterogeneity in MSI-H, particularly in the sporadic and hereditary settings, (c) molecular mechanisms underlying the MSI(+) phenotypes, and (d) relationships between the MSI(+) and CIN phenotypes. Several molecular mechanisms may underlie repeat instability in eukaryotic cells. The relationship between MSI and defective MMR may be more complicated than has been suspected. The role of MMR deficiency in tumorigenesis in the digestive tract appears to be diverse and is not simple, even in the colorectum. PMID- 16341481 TI - Expired gas analysis during exercise testing pre-pneumonectomy. AB - PURPOSE: Expired gas analysis has enabled the successful prediction of postoperative complications in patients undergoing thoracic esophagectomy. We conducted this study to determine whether preoperative expired gas analysis during exercise testing can help identify patients at high risk of postoperative complications after pneumonectomy. METHODS: We measured the vital capacity, percent vital capacity, forced expiratory volume in 1.0 s, percent forced expiratory volume in 1.0 s, maximum oxygen uptake per minute, anaerobic threshold, arterial partial pressure of oxygen, and arterial partial pressure of carbon dioxide in 27 patients scheduled to undergo pneumonectomy. Group A consisted of 18 patients without postoperative cardiopulmonary complications and group B consisted of 9 patients with postoperative cardiopulmonary complications. We compared preoperative cardiopulmonary data between these two groups. RESULTS: Postoperative cardiopulmonary complications developed in 9 of the 27 patients (33.3%), 3 (11%) of whom died. The maximum oxygen uptake and the anaerobic threshold were significantly higher in group A than in group B (P < 0.05), whereas spirometric pulmonary function testing and arterial blood gas analysis showed no intergroup differences. CONCLUSION: Expired gas analysis during exercise testing can help identify patients at high risk of postoperative cardiopulmonary complications after pneumonectomy. PMID- 16341482 TI - Interleukin-8, cyclo-oxygenase-2, and trefoil factor family 1 gene expression and their association with Helicobacter pylori infection in the remnant stomach. AB - PURPOSE: The risk factors for secondary stomach carcinogenesis after distal gastrectomy have not been evaluated in detail. METHODS: Using gastrointestinal endoscopy, we examined 112 patients who had undergone gastrectomy. Biopsy specimens were taken from the stoma and the upper corpus mucosa in the remnant stomach to examine the associations among Helicobacter pylori (H.pylori) infection, bile reflux, and the expressions of interleukin-8 (IL-8), cyclo oxygenase-2 (COX-2), and trefoil factor family 1 (TFF1) genes in the stomach mucosa. RESULTS: The IL-8 levels in the corpus mucosa were significantly higher in the H.pylori-positive patients than in the H.pylori-negative patients (P = 0.015). The IL-8 levels were significantly higher in the stomal mucosa than in the corpus mucosa in the H.pylori-positive patients (P = 0.047). The COX-2 levels in the corpus mucosa tended to be higher in the H.pylori-positive patients, but these levels were not significantly different in the stoma mucosa. The COX-2 levels in the corpus were significantly higher after Billroth II (BII) anastomosis than after Billroth I (BI) anastomosis (P = 0.041). TFF1 expression in the stoma was higher in the H.pylori-positive patients than in the H.pylori negative patients, but the difference was not significant. CONCLUSIONS: Both H.pylori infection and bile reflux increased IL-8 levels after BI anastomosis. Furthermore, COX-2 levels were higher after BII than after BI anastomosis. These indicators will become useful not only as biomarkers to predict the degree of inflammation in the stomach mucosa, but also as surrogate biomarkers to predict the risk of secondary stomach carcinogenesis in the remnant stomach mucosa. PMID- 16341483 TI - Association between recurrence of hepatocellular carcinoma and alpha-fetoprotein messenger RNA levels in peripheral blood. AB - PURPOSE: Intra- and extrahepatic recurrence is common, even after curative resection for hepatocellular carcinoma (HCC), suggesting preoperative or intraoperative tumor cell dissemination. Reverse transcription - polymerase chain reaction (RT-PCR) for alpha-fetoprotein (AFP) is used to detect circulating liver cancer cells. We previously developed a quantitative method that allows estimation of the AFP mRNA level by real-time PCR. In the present study, we used this method to measure the AFP mRNA level before and after resection of HCC, then correlated the findings with various clinicopathological characteristics and prognosis. METHODS: We prospectively examined peripheral blood samples from 38 patients with HCC, and bone marrow aspirate from 25 of these patients. As a control, we examined bone marrow from 20 patients with benign diseases. The follow-up period ranged from 32 to 66 months. Real-time RT-PCR was used to detect AFP mRNA levels in the samples. RESULTS: AFP was expressed in 9 (23.7%) of the 38 peripheral blood samples. The detection of AFP mRNA was significantly correlated with extrahepatic metastasis after primary surgery, and a shorter disease-free survival time (P = 0.0245 each). Bone marrow samples were defined as positive if they expressed AFP mRNA at levels higher than the maximum expressed level in the controls, because only 1 (5%) of the 20 control bone marrow samples had low AFP mRNA expression. Using this cutoff level, 12 (48%) of the 25 patients with HCC had positivity for AFP mRNA. The results of bone marrow RT-PCR did not correlate with the clinocopathological characteristics of prognosis. CONCLUSIONS: Using real-time PCR to measure the AFP mRNA level in blood, but not bone marrow, could be useful for predicting postoperative tumor recurrence. PMID- 16341484 TI - Long-term safety of autotransfusion during hepatectomy for hepatocellular carcinoma. AB - PURPOSE: To evaluate the long-term safety of autotransfusion (AT) in hepatectomy for hepatocellular carcinoma (HCC). METHODS: Between 1988 and 1989, 46 patients with HCC underwent hepatectomy with AT (group 1). For a comparison, we matched 50 patients with HCC who underwent hepatectomy, and received homologous but not autologous blood (group 2). The 10-year cumulative survival curves and cancer free curves of the two groups were examined, and the pattern of recurrence was compared. RESULTS: Group 1 had a significantly higher cumulative 10-year survival rate than group 2, at 20% vs 8%, respectively (P < 0.05). Among the patients who underwent curative resection, those in group 1 had significantly better cumulative survival and cancer-free survival rates than those in group 2, at 27% vs 11% (P < 0.05) and 13% vs 0% (P < 0.05), respectively. Among the patients with stage I-II HCC, those in group 1 had significantly better cumulative survival and cancer-free survival rates than those in group 2, at 30% vs 5% (P < 0.01) and 20% vs 5% (P < 0.05), respectively. However, the rates were similar among patients with stage III-IV disease in both groups. The pattern of recurrence in the two groups was similar. CONCLUSION: Autotransfusion promoted survival in patients undergoing hepatectomy for stage I or II HCC. PMID- 16341485 TI - Tension-free hernioplasty versus conventional hernioplasty for inguinal hernia repair. AB - PURPOSE: To show the effectiveness of tension-free hernioplasty for inguinal hernia repair. METHODS: We studied 106 patients who underwent inguinal hernia repair, as conventional hernioplasty in 52 and as tension-free hernioplasty in 54. We analyzed the operation time, postoperative complications, pain, time to resume daily activities, and frequency of recurrence in the short and long term. RESULTS: The average age of the patients was 46.2 years. The operation time was significantly shorter in the tension-free group than in the conventional group, at 33 +/- 11.1 versus 49 +/- 8.8 min, respectively (P < 0.05). The overall complication rate was 10%, being 1.5% in the tension-free group and 13% in the conventional group (P = 0.4). The visual-analogue pain scores after surgery were lower in the tension-free group than in the conventional group (P = 0.01). Patients in the tension-free group returned to their normal activities sooner than those in the conventional group (P < 0.05). CONCLUSIONS: Tension-free hernioplasty resulted in less pain and allowed patients to return to their daily activities sooner than conventional hernioplasty. PMID- 16341486 TI - Seprafilm does not aggravate intraperitoneal septic conditions or evoke systemic inflammatory response. AB - PURPOSE: To determine whether Seprafilm aggravates the systemic inflammatory response and adversely affects the outcome of postoperative intraperitoneal septic conditions. METHODS: We retrospectively analyzed the relationship between the intraperitoneal placement of Seprafilm and the rate of intraperitoneal septic complications in 278 consecutive patients. Experimentally, 40 rats were subjected to laparotomy with or without the intraperitoneal placement of Seprafilm. Bacterial peritonitis was induced by cecal ligation and puncture (CLP) and the serum cytokine levels were measured. RESULTS: Seprafilm did not increase the rate of septic complications or aggravate inflammatory responses in patients with or without postoperative intraperitoneal inflammatory complications. Experimentally, there was no significant difference between the serum inflammatory cytokine levels after CLP with or without Seprafilm. CONCLUSIONS: Seprafilm did not adversely affect postoperative inflammatory response or clinical outcomes, even in patients with intraperitoneal septic complications. PMID- 16341487 TI - Decrease in the incidence of Buerger's disease recurrence in northern Thailand. AB - PURPOSE: Buerger's disease is a peripheral arterial occlusive disease (PAOD), which occurs mainly in young male smokers. It is common in Asia, the Middle East, and Eastern European countries, but rare in North America and Western European countries. We investigated the changes in the prevalence and recurrence of Buerger's disease at a referral hospital in northern Thailand. METHODS: We conducted a retrospective study of patients with Buerger's disease admitted to Chiang Mai University Hospital between January 1988 and December 2002. Buerger's disease was diagnosed according to Shionoya's clinical criteria. RESULTS: We evaluated 78 patients with Buerger's disease, with a collective total of 108 admissions. Forty-one patients were admitted for initial treatment, and 37 patients had a history of treatment and were admitted for worsening of Buerger's disease. Between 1988 and 1995, 23 patients were admitted with newly diagnosed Buerger's disease, 52 were admitted because of Buerger's disease exacerbation, and 372 were admitted for a PAOD; whereas between 1996 and 2002, the respective numbers were 18, 15, and 632 (2.9 +/- 1.9/year vs 2.6 +/- 1.3/year, P = 0.25; 6.5 +/- 1.1/year vs 2.1 +/- 1.1/year, P = 0.001; 54.1 +/- 6.7/year vs 81.6 +/- 5.2/year, P < 0.0001). Of the 78 patients with Buerger's disease, 76 were men and 2 were women, with a mean age at onset of 34.1 +/- 5 years. The predominant symptom on admission was burning pain in the feet and hands in 75%, digital gangrene in 74%, and a digital ulcer in 43.5%. CONCLUSIONS: The incidence of recurrence of Buerger's disease seems to be decreasing in our institution, whereas the incidence of this disease and its recurrence both seem to be decreasing in Western countries and Japan. PMID- 16341488 TI - Polidocanol at different concentrations for pleurodesis in rats. AB - PURPOSE: We previously found that 0.5% polidocanol was more effective than tetracycline for pleurodesis in rats. Thus, we conducted the present study to evaluate the efficacy of different concentrations of polidocanol for pleurodesis in rats. METHODS: We divided 54 albino Wistar rats into six groups. Groups 1, 2, and 3 were given isotonic saline, 35 mg/kg tetracycline, and 0.6 mg of diluted polidocanol, respectively, being the daily recommended dose for humans. Groups 4, 5, and 6 were given 0.5%, 1%, and 2% polidocanol, respectively. All solutions were given intrapleurally in a volume of 0.5 ml. We examined the rats for macroscopic pleural adhesions and compared the mean values of macroscopic scoring among the six groups. RESULTS: The rats given polidocanol and tetracycline had significantly more adhesions than the control group, and polidocanol at concentrations of 0.5%, 1%, and 2% was more effective for pleurodesis than tetracycline. The diluted polidocanol was not more effective than tetracycline. There was no difference between the effects of the 0.5% and 1% concentrations, but the 2% polidocanol group had significantly more adhesions than the other groups. CONCLUSIONS: Polidocanol at concentrations of 0.5%, 1%, and 2% was a more effective sclerosing agent than tetracycline for pleurodesis. While 2% polidocanol was the most efficient sclerosing agent, the daily maximum recommended dose of polidocanol for humans was not more effective than tetracycline. PMID- 16341489 TI - Cervicomediastinal thymic cyst: report of a case. AB - Congenital thymic cysts are rare. Consequently, they are often misdiagnosed and not included in the preoperative differential diagnosis of neck masses. We report the case of a 7-year-old boy with a large cervicomediastinal thymic cyst to increase the awareness of this unusual entity. We discuss the clinical features, presentation, and pathogenesis of thymic cysts. PMID- 16341490 TI - Aneurysmal bone cyst of the rib: report of a case. AB - Aneurysmal bone cysts (ABC) are rare, especially in the ribs. This lesion is considered to be a benign process, which is usually found in children and young adults. This case, however, concerns a 71-year-old patient, the oldest patient hitherto reported in Japan, and also as old as in any other published case report worldwide. Although positron emission tomography (PET) demonstrated an uptake at the lesion, two biopsies could not reveal a precise preoperative diagnosis. The clinical manifestations, etiology, pathology, and current treatment are briefly discussed. Due to difficulties in the preoperative diagnosis, and to avoid recurrence, a total excision is the recommended treatment for ABC. PMID- 16341491 TI - Total anomalous pulmonary venous connection with a ductus arteriosus aneurysm in a neonate: report of a case. AB - We performed successful emergency surgery on a 1-day-old infant to repair a total anomalous pulmonary venous connection (TAPVC) and a ductus arteriosus aneurysm (DAA) diagnosed by two-dimensional echocardiography. We anastomosed the common pulmonary venous chamber to the left atrium and ligated the pulmonary end of the DAA. Echocardiography and cardiac catheterization done 3 months later showed that the DAA had regressed. To the best of our knowledge, this is the first report describing the successful repair of a supracardiac TAPVC with a DAA. PMID- 16341492 TI - Surgical treatment of a superior sulcus tumor in a patient with idiopathic thrombocytopenic purpura: report of a case. AB - We herein report a resection of a superior sulcus tumor in a patient with idiopathic thrombocytopenic purpura. A resection of the left upper lobe of the lung, left subclavian artery, and left first to third ribs, as well as a reconstruction of the left subclavian artery, were performed. Postoperative hemorrhaging was controlled due to preoperative high-dose intravenous immunoglobulin therapy and a platelet transfusion both during and following surgery. The resected tumor was diagnosed to be a pulmonary pleomorphic carcinoma, which was pathologically determined to be T3N0M0-Stage 2B. The patient remained in good condition for 20 months following the surgery; however, he eventually died due to bone metastases. PMID- 16341493 TI - Spontaneous regression of hepatocellular carcinoma: report of a case. AB - A spontaneous regression of hepatocellular carcinoma is an extremely rare phenomenon. A 69-year-old Japanese man with hepatitis C virus-related chronic hepatitis presented with a liver tumor. We diagnosed the tumor to be hepatocellular carcinoma in the course of spontaneous regression, by imaging studies and changes in the tumor markers. Because the possible presence of viable cancer cells could not be ruled out, we recommended surgery. He refused all treatments at first, but finally agreed to undergo surgery about 10 months after presentation. A hepatectomy was performed. Histologically, no viable tumor cells were found. In our case, the vascularity of the tumor according to the imaging findings was followed up during the clinical course. The patient is now doing well and without any evidence of recurrence at 37 months after surgery. PMID- 16341494 TI - Improved quality of life and unchanged magnetic resonance brain imaging after living donor liver transplantation for late-onset ornithine transcarbamylase deficiency: report of a case. AB - We report the case of a 7-year-old girl with ornithine transcarbamylase deficiency whose quality of life (QOL) improved greatly after a living donor liver transplantation (LDLT). Ornithine transcarbamylase deficiency had been diagnosed when she was 2 years old and she finally underwent LDLT, with her father as the donor, when she was 7 years old. The patient had suffered episodes of hyperammonemic encephalopathy ranging from lethargy to coma, treated by hemodialysis twice before LDLT, and her intelligence quotient was borderline for her age. Preoperative magnetic resonance imaging (MRI) showed an atrophic area in the subcortical white matter of the frontal lobe. After LDLT, the patient suffered acute rejection with hyperamylasemia, but not hyperammonemia. Postoperative MRI and quantitative MR spectroscopy showed no changes in the subcortical lesion. She has been followed up carefully for 16 months and has had no further complications or any sign of hyperammonemia. PMID- 16341495 TI - Intraoperative endoscopic resection of left ventricular tumors. AB - Two cases involving patients who underwent a successful endoscopic resection of a left ventricular tumor are presented herein. One was an 82-year-old woman with a left ventricular papillary fibroelastoma, who underwent previous coronary artery bypass grafting. In an attempt to make the procedure less invasive, we used an endoscope. With a full sternotomy, cardiopulmonary bypass, and cardioplegic protection, the endoscope was inserted into the left ventricular cavity through the mitral valve. The other patient was a 63-year-old man with left ventricular papillary fibroelastoma, in whom we performed an endoscopic transaortic resection. The endoscope provided an excellent view, and the tumors were easily extracted in both cases without any complications. PMID- 16341496 TI - Evaluation of the predictors of choledocholithiasis: comparative analysis of clinical, biochemical, radiological, radionuclear, and intraoperative parameters. PMID- 16341497 TI - Childhood predictors of externalizing and internalizing problems in adolescence. A prospective follow-up study from age 8 to 16. AB - The aim of this study was to assess the childhood predictors of externalizing and internalizing symptoms in adolescence in an epidemiological sample. Behavior ratings were obtained from 609 children at two time-points, accounting for 71% of the target sample. At age 8, children were evaluated with parental and teacher Rutter scales, and with the Child Depression Inventory (CDI), and at age 16 with the Child Behavior Checklist. Evaluations by all informants had a unique contribution to later outcome. In multivariate analysis, among boys, parental reports of hyperactivity independently predicted externalizing problems and teacher reports of hyperactivity independently predicted internalizing problems. Teacher reports of conduct problems independently predicted externalizing problems among both boys and girls. Furthermore, parent reports of emotional problems independently predicted internalizing problems among both boys and girls. Children's own reports of internalized distress measured with CDI predicted a high level of internalizing problems among girls. Perceived need of treatment was the strongest predictor for outcome among girls. Change in family structure (e. g., divorce or remarriage) during follow-up independently predicted externalizing and internalizing problems among boys. The study supports the findings from earlier studies showing that the stability of behavior problems from childhood to adolescence is substantial. This implies a need for early recognition and initiation of treatment efforts. PMID- 16341498 TI - Chronic fatigue syndrome of childhood. Comparative study with emotional disorders. AB - OBJECTIVE: To examine clinical specificity in chronic fatigue syndrome (CFS) of childhood, by comparing clinical features in childhood CFS and in emotional disorders (ED). METHOD SAMPLE: 28 children with CFS; 27 with ED. MEASURES: History of disorder; K-SADS psychiatric interviews; self-esteem and physical symptoms questionnaires; premorbid history, behavioural and personality assessments. RESULTS: There were high levels of comorbid emotional disorders in children with CFS, and the two groups were comparable on self-esteem, but CFS children endorsed more fatigue and other somatic symptoms. The two groups were comparable on age at illness onset, but parents of children with CSF reported more biological illness precipitants, more pre-morbid recurrent medical problems and infections. The CFS group had fewer pre-morbid psychological problems and less psychiatric comorbidity than the ED group. CONCLUSION: There is considerable clinical overlap between CFS and ED of childhood, but there are also differences in clinical presentation between these disorders. PMID- 16341499 TI - Childhood onset neuropsychiatric disorders in adult eating disorder patients. A pilot study. AB - BACKGROUND: Autism spectrum disorders (ASD) have been suggested to be overrepresented in anorexia nervosa. This study aimed to explore the comorbidity of ASD and other childhood onset neuropsychiatric disorders (COND) [attention deficit/hyperactivity disorder (AD/HD) and tic disorders] in a group of severe eating disorder (ED) patients. METHOD: Thirty female ED patients from a specialist hospital clinic were examined on measures tapping into COND and personality disorders. RESULTS: In our group of longstanding ED, 53% had at least one COND diagnosis; 23% had ASD, 17% had AD/HD, and 27% had a tic disorder. CONCLUSIONS: These preliminary data suggest that COND may be common in patients with severe ED and should be kept in mind when treating these patients. PMID- 16341500 TI - Mental and somatic health in a non-clinical sample 10 years after a diagnosis of encopresis. AB - The aim of this study was to assess the relation between the diagnosis of encopresis at 8 and 10 years of age, and mental and somatic health 10 years later. The importance of type of encopresis (primary or secondary) at 8 years was also studied. Subjects were a non-clinical encopretic sample (N=73) and control subjects (N=75) [2]. Seven assessment variables from conscription surveys provided information about mental and somatic health status at 18 years of age. Former encopretics (n=66) did not differ significantly from the controls (n=67) at 18 years of age, although there were consistent, small negative differences. The boys who at 10 years of age had still been encopretic did not differ significantly at 18 years of age from the boys who at 10 years had recovered from encopresis, and the signs indicating the small differences varied. For former primary and secondary encopretic boys, there were two significant differences, the men in the secondary group being more often exempted from conscription than the primary group and the control cases. The results indicate that boys with non clinical encopresis show only small, if any, mental and somatic disturbances at the beginning of adulthood. Comprehensive investigations of encopretic patients are recommended as important clinical problems, in addition to encopresis, might be present. PMID- 16341501 TI - School factors and the emergence of depressive symptoms among young Norwegian adolescents. AB - The aims of this study were to examine correlates of self-reported depressive symptoms within the school area, and possible predictors of change in depressive symptom levels over a 1-year period in a large representative sample of 12 to 15 year-old adolescents in central Norway. The school variables were: school stress, class wellbeing, teacher support, and grades. Possible confounding factors were age, SES, ethnicity, and parental divorce. We also examined the strengths of these relationships. In the cross-sectional multivariate analyses of the whole sample, at time-point one (T1), all four school factors were independently associated with depressive symptom levels at small to moderate levels. In the longitudinal multivariate analyses of the whole sample, self-reported depressive symptom levels at time-point two (T2) were predicted by depressive symptom levels at T1, gender and teacher support. For girls, three of the school variables significantly predicted depressive symptom levels at T2, while this was not the case for boys. Prevention in schools might be of importance. PMID- 16341502 TI - Life events and suicidality in adolescents with schizophrenia. AB - OBJECTIVE: Suicide is the leading cause of premature death in patients with schizophrenia. Studies have shown a weaker association between suicidal behavior and stressful life events in schizophrenic than in nonschizophrenic subjects. The aim of the present study was to further investigate the complicated relationship of suicide attempts and life events in adolescent schizophrenic patients. METHODS: Forty adolescents with a diagnosis of schizophrenia, including 20 who had attempted suicide and 20 who had not, were compared with 20 age-matched subjects with no psychiatric history. The instruments used for the assessment were the Life Events Checklist, the Suicidal Risk Scale, the Sexual Abuse in Childhood Questionnaire, and the Beck Depression Inventory. RESULTS: Control subjects reported fewer life events in general, and fewer negative events, events of sexual abuse, and events associated with impaired family functioning than the schizophrenic patients. Within the schizophrenic group, the suicidal patients reported fewer life events than the nonsuicidal patients, but there was no difference between the groups in the number of negative or sexual-abuse events. However, the proportion of negative life events out of total life events was higher in the suicidal group, and their perceived impact was stronger. Levels of depression and suicidality were higher in the suicidal schizophrenic patients than in the nonsuicidal patients. CONCLUSIONS: In adolescent patients with schizophrenia, suicidal behavior is associated less with the number of life events and more with their perception of the events as negative and the impact of these events on the individual. PMID- 16341503 TI - The test-retest reliability and screening efficiency of DISC Predictive Scales version 4.32 (DPS-4.32) with Chinese children/youths. AB - INTRODUCTION: The DISC Predictive Scales-4.32 (DPS-4.32) were short-forms of the Diagnostic Interview Schedule for Children-IV (DISC-IV). METHOD: The psychometric properties of DPS-4.32 were tested on two Chinese samples: a clinic sample of 83 children/youths, and a community sample of 541 youths. RESULTS: Both Parent and Youth DPS-4.32 exhibited good-to-excellent test-retest reliability. Their screening efficiency was examined respectively against DSM-IV diagnoses of the full-length Parent and Youth DISC-IV. Results indicated large AUCs (Area under Receiver-Operating Characteristic Curve), as well as high sensitivity, specificity, and negative predictive values, supporting the capability of DPS 4.32 to differentiate cases from noncases. CONCLUSION: DPS-4.32 held promise as efficient short-forms of DISC-IV, screening DSM-IV diagnoses. They were also applicable to the Chinese population, demonstrating their cross-cultural applicability. PMID- 16341504 TI - Can we--and should we--have a Europsychiatry for Children and Adolescents? The work of the UEMS Section and Board for Child and Adolescent Psychiatry/Psychotherapy. AB - The Union of European Medical Specialists (UEMS) paves the way for harmonisation of training and free movement of medical doctors within the European Union. For more than 10 years, Child and Adolescent Psychiatry has been a distinct specialty at this European level--separate from Adult Psychiatry and Pediatrics. The article gives detailed information on the background of the section/board of Child and Adolescent Psychiatry/Psychotherapy (CAPP), training issues including the Training Log Book and the recent situation as well as future perspectives of Continuous Professional Development (CPD) in CAPP, all of which influence the corner stones of CAPP and its delineation from other medical and non-medical organisations. Child and Adolescent Psychiatry as the application of trained specialist medical practice to mental illnesses and psychological disorders in children and young people up to the age of about 18 years reflects more and more the growing research advances of the last years within the field with more progress to come. On the other hand, shortcomings of patient provision in Europe still have to be resolved and CAPP may help to do so. PMID- 16341507 TI - Sharing instrumentation globally. PMID- 16341505 TI - Attention and behavioural problems of Finnish adolescents may be related to the family environment. AB - BACKGROUND: Adolescents' attention and behavioural problems in relation to the family environment were studied in the Northern Finland 1986 Birth Cohort. METHOD: Fifteen-year-old adolescents (N=6888) completed the Youth Self-Report (YSR) questionnaire and their parents (N=6643) completed the Strengths and Weaknesses in ADHD and Normal Behaviours (SWAN) questionnaire. The ratings were compared in relation to gender and family characteristics. RESULTS: Girls scored higher than boys on the YSR attention problems subscale (means 4.61 vs. 3.41), the rule-breaking behaviour subscale (means 4.25 vs. 3.69) and the aggressive behaviour subscale (means 7.18 vs. 5.63). Parents reported more SWAN attention problems in their sons than in their daughters. Living in an other than intact family increased YSR and SWAN attention problems and YSR behavioural problems in both genders. Adolescents living in very large families (11-19 children) had least attention and behavioural problems. CONCLUSIONS: Attention and behavioural problems seem to be common among adolescents in Finland. For both genders, living in other than intact families increases especially behavioural problems. Additionally, a very large family seems to be a protective factor against those problems. PMID- 16341508 TI - Time-resolved chemiluminescent analysis of hydralazine in pharmaceuticals. AB - A new analytical method is proposed for determination of hydralazine (HZ) in pharmaceuticals--measurement of the chemiluminescence (CL) emitted after reaction with phosphoric-acidified KMnO4. The novelty of this method is the recording of the whole CL-time profile. Such a recording is possible by use of a CL-detector operating in tandem which enables the reactants to be mixed in the measurement cell only and, therefore, the CL is reaction monitored from beginning. At the precise time the pump is stopped signal recording is triggered and so CL evolution is recorded completely. The optimum chemical conditions for the determination were 0.8 mol L(-1) formaldehyde, 0.3 mmol L(-1) KMnO4, 4.0 mol L( 1) H3PO4, and a total flow of 0.37 mL s(-1). Two calibration graphs were plotted, CL intensity and area under the profile curve against HZ concentration. Exhaustive statistical analysis provided very interesting results, for example, accordance with Clayton's theory, detection limit below 0.2 microg mL(-1), and linear calibration ranges from 0.2 to 5.0 microg mL(-1). This method was successfully applied to the determination of HZ in pharmaceuticals. Because they are usually formulated in association with diuretics and beta-blockers, the method was used for analysis of HZ in pharmaceuticals that contained either HZ only or HZ with other hypotensive substances. Obtained and nominal content were approximately the same and experimental Student t values indicated there were no significant differences between the values. PMID- 16341509 TI - Electrochemical properties and square-wave voltammetric determination of pravastatin. AB - The electrochemical reduction and adsorptive voltammetric behaviour of pravastatin have been studied by means of cyclic and square-wave voltammetry at a hanging mercury-drop electrode in electrolytes of different pH. Within the entire pH range (2.0-9.0) in Britton-Robinson buffer, pravastatin gave rise to a single voltammetric peak in the potential interval from -1.22 to -1.44 V, depending on pravastatin concentration. It was found that the reduction of pravastatin proceeds via a relatively stable intermediate, which is transformed to the final electroinactive product by a coupled chemical reaction or can be re-oxidized back to pravastatin. The rate of chemical transformation is controlled by the proton concentration. The electrode mechanism has the properties of a surface redox reaction. A sensitive analytical method for trace analysis of pravastatin based on the adsorptive stripping technique has been developed. The calibration plot was linear in the range 8x10(-8)-5x10(-7) mol L(-1). Application of the square wave voltammetric method to determination of pravastatin in a pharmaceutical dosage form, without sample pretreatment, resulted in acceptable deviation from the stated concentration. PMID- 16341510 TI - Ultraviolet photolysis of urine for suppression of color quenching prior to liquid scintillation counting of tritium. AB - In order to reduce the color quenching in the measurement of tritium in urine by liquid scintillation counting , UV irradiation was applied to decompose the organic substances in the sample. Urine was decolorized under UV irradiation in the presence of hydrogen peroxide. As a result, color quenching was considerably suppressed and higher counting efficiency of tritium was obtained. This UV treatment made it possible to increase the urine content in the sample from 2 to 40% (v/v) without significant decrease of counting efficiency. Either higher sensitivity or shorter analysis time was achieved in the tritium measurement by the augmentation of urine content. When the measurement time was 30 min, the detection limit of tritium defined as 3s was 0.03 Bq/ml. At the expense of some sensitivity (set at a detection limit of 0.3 Bq/ml), the measurement time was shortened to 0.5 min. These results will make a great improvement to routine tritium monitoring as well as to emergency monitoring in mass tritium exposure. PMID- 16341511 TI - [Iliac ureteral fistulas. Diagnosis and management of an increasing problem]. AB - Iliac-ureteral fistulas (IUF) are a rare but potential life threatening event and an important cause of gross hematuria. We report on three cases of IUF. In all cases, prior chronic ureteral stenting, extended pelvic surgery or pelvic irradiation had been performed. Diagnosis was confirmed with angiography in one case, in the others a CT scan revealed the IUF. Treatment included surgical exploration with local reconstruction, extra-anatomical bypass and nephrectomy with arterial patch repair. The increasing incidence of IUF is a consequence of an increasing number of advanced and extended pelvic operations, radiation therapy and long-term ureteral stenting. Diagnosis should be made by provocative angiography or CT. Treatment options vary depending on the site and morphology of the local situation, but morbidity and mortality is still high due to delayed adequate diagnosis and treatment. A conclusive algorithm should be followed for the successful management of IUF. PMID- 16341512 TI - [Management of erectile dysfunction after radical prostatectomy. Urologists' assessment vs patient survey responses]. AB - INTRODUCTION: The aim of this national study was to evaluate ED management after RPX (without any postoperative adjuvant therapy or tumor relapse) from the patient's view compared to the urologist's view. MATERIAL AND METHODS: In May 2003 we queried 1063 urologists and 801 patients following radical prostatectomy without adjuvant therapy. They were asked about preserved potency without erectile aid, existing wish for ED therapy, recommended or tested erectile aid (oral, transurethral, intracorporal, vacuum constriction device[VCD], penile implant) as well as the long-term use. Return rate: patients 80.1%, urologists 26.7%. RESULTS: According to the urologists' view 9.1% of their affected patients were potent postoperatively without a device, but according to the polled patients only 4.7%. The wish to be treated for erectile dysfunction existed in the urologists' opinion in 46.1% of their patients, while they considered that 44.8% had no wish for treatment. On the other hand, 59.3% of the patients would like to be treated and only 28.5% did not want any kind of treatment. Regarding the long-term use of therapy for ED, the urologists thought that 26.1% of their patients did not receive therapy for the problem, and 69.7% of the patients stated they received no long-term therapy. Only 30.3% of the patients confirmed long-term therapy, while the urologists thought that 73.9% of the patients used an erectile aid. Definite therapy in the urologists' opinion involved: oral medication in 38.4%, MUSE in 3.6%, (SKAT) in 37.3%, VCD in 20.4%, and a prosthesis in 0.3%. Indeed 19.8% of the patients used oral medication, 1.7% MUSE, 26.7% SKAT, 50.9% VCD, and 0.9% penile implant. Considering the satisfaction of patients, urologists thought that 46.2% of the patients were satisfied with their treatment of ED, but only 28.9% of the patients were actually satisfied themselves. CONCLUSIONS: The comparison of patients' and urologists' views shows a clearly different description of the ED situation after RPX. The proportion of patients with a wish for treatment and the proportion of dissatisfied patients are much higher from the patients' view. This demonstrates an undertreatment of ED patients after RPX, which should also be taken into account under the current changes in the German health care system. PMID- 16341513 TI - Identification of interleukin-2 for imaging atherosclerotic inflammation. PMID- 16341514 TI - Primer on molecular imaging technology. AB - A wide range of technologies is available for in vivo, ex vivo, and in vitro molecular and cellular imaging. This article focuses on three key in vivo imaging system instrumentation technologies used in the molecular imaging research described in this special issue of Eur J Nucl Med Mol Imaging: positron emission tomography, single-photon emission computed tomography, and bioluminescence imaging. For each modality, the basics of how it works, important performance parameters, and the state-of-the-art instrumentation are described. Comparisons and integration of multiple modalities are also discussed. The principles discussed in this article apply to both human and small animal imaging. PMID- 16341515 TI - [Adverse drugs reactions: diagnosis and assessment]. AB - Adverse drug reactions (ADR) occur in about 5% of all pharmacologically treated patients. Between 2% and 20% of all hospital admissions are caused by ADR, and approximately 10% of all hospitalized patients experience ADR during their hospital stay. Several thousand patients die due to ADR in Germany each year. ADR associated drugs come predominantly from the class of non-steroidal antiinflammatory drugs, anticoagulants, acetylsalicylic acid and cardiovascular drugs. Most ADR cases present as gastrointestinal bleeding and adverse cardiovascular effects. Apart from this, one or more drugs are withdrawn from the market each year because of unwanted but mostly rare side effects. In recent years the most prominent cases were rofecoxib, cisapride and cerivastatin. Physicians in Germany are obliged to report ADR. A substantial proportion of ADR, however, is not reported because it is deemed to be either too well known or the association between the drug and the adverse effect is too doubtful. In some cases, histopathological findings are needed to determine the diagnosis of ADR. Accordingly, physicians should inform the pathologist whether an ADR is suspected and which drugs may be responsible. PMID- 16341516 TI - [Regressive changes after short-term neoadjuvant antihormonal therapy in prostatic carcinoma: the value of Gleason grading]. AB - Although neoadjuvant, antihormonal therapy does not lead to an improvement in the outcome of prostatic carcinoma it is still used in the short-term in a subset of patients. Here we report the regressive changes due to this short-term treatment and analyse the impact on Gleason grading. The most frequent regressive changes in 82 tumors treated short-term were determined and quantified. The results were compared to a matched control group and also to the preoperative needle biopsies.A steep increase in regressive changes was observed within the first 4 weeks. After this point, changes increased only mildly. Within the first 2 weeks of treatment no significant changes compared to control tissue were present. Compared to the preoperative needle biopsies, pretreated tumors showed a significant upgrading. After 2 weeks of neoadjuvant antihormonal therapy, regressive changes are so great, that Gleason grading can no longer be recommended. PMID- 16341517 TI - [Fluorosis--a forgotten entity. Case report of a woman with coxarthrosis and newly diagnosed fluorosis]. AB - The clinical manifestation of fluorosis has become rare over the past years. Although the use of fluoride medication in osteoporosis therapy remains controversial, past study results have led to a reduction in fluoride prescriptions. Several studies have shown minor biomechanical properties of newly built woven bone compared to original bone. Despite new prescription protocols, fluoride therapy should not be disregarded in the anamnesis of osteoporosis patients. In addition to conventional diagnostics in fluorosis, new techniques such as microanalysis and micro-CT-analysis show a diagnostic benefit. In this case, the edx-microanalysis results show an F concentration of over 1.0 wt% in bone. The ratio of bone to tissue volume, evaluated by micro-CT, is clearly elevated at 46% BV/TV. The histopathological preparation of the femoral head has made the possible effects of fluoride medication on bone visible and quantifiable. A direct causal relationship between coxarthrosis and fluoride medication, found both in our patient as well as in the literature, has not been demonstrated. In order to better understand the broad effects of fluoride medication in combination with coxarthrosis more studies are needed. PMID- 16341518 TI - [Intramuscular nodular fasciitis--a clinicopathological study with emphasis on myogenic giant cells]. AB - Nodular fasciitis is a frequently occurring pseudosarcomatous fibrous proliferation which usually affects the subcutaneous tissue. In the present study, we investigated tissue specimens from 26 patients with intramuscular nodular fasciitis. The median age of patients was 31 years, with the tumors occurring mainly in the extremities (11 patients) and the trunk (11 patients). These had an average size of 2.6 cm. Males predominated (male:female 9:4). Histological examination revealed a proliferation of myofibroblasts (positive for smooth muscle-actin) embedded in a myxoid or fibrous stroma. In 25/26 cases, myogenic giant cells were observed (positive for desmin, negative for smooth muscle actin and CD 68). Less frequently (11/26 patients), osteoclastic giant cells were found (positive for CD 68, negative for desmin and smooth-muscle actin). In intramuscular nodular fasciitis, myogenic giant cells are encountered more often than previously described. They are relevant for the differential diagnosis of sarcoma and fibromatoses, since they can be misinterpreted as neoplastic giant cells. However, the uniformity of the myofibroblastic proliferate and the myogenic phenotype of the giant cells without proliferative activity (no mitoses, negativity for Ki-67) lead to a correct diagnosis. PMID- 16341519 TI - Real-time PCR for detection of Brucella spp. DNA in human serum samples. AB - Presented here are the results of an evaluation of an in-house real-time PCR assay for the rapid and specific diagnosis of human brucellosis. The assay was based on direct amplification from serum samples of a 169-bp portion of bcsp31, a gene found in all Brucella species and biovars. Species specificity and selectivity of this real-time PCR assay were evaluated using genomic DNA from 15 Brucella strains and 42 non-Brucella strains, and the results were 100%. Among 17 culture-proven brucellosis patients, sera from 11 gave a positive amplification signal, corresponding to a sensitivity of 64.7%. In contrast, negative results were obtained for all sera from 60 control patients, corresponding to a specificity of 100%. The results indicate this test is well adapted for definite confirmation of brucellosis cases, when Brucella cultures remain sterile and serological tests demonstrate the presence of cross-reacting antibodies against Brucella sp. and Yersinia enterocolitica O:9 antigens. PMID- 16341520 TI - New strains of community-acquired methicillin-resistant Staphylococcus aureus with Panton-Valentine leukocidin causing an outbreak of severe soft tissue infection in a football team. PMID- 16341521 TI - Substitution of heat from exercise and digestion by ducks diving for mussels at varying depths and temperatures. AB - Diving birds can lose significant body heat to cold water, but costs can be reduced if heat from exercising muscles or the heat increment of feeding (HIF) can substitute for thermogenesis. Potential for substitution depends jointly on the rate of heat loss, the rate of heat produced by exercise, and the level of HIF. To explore these interactions, we measured oxygen consumption by lesser scaup ducks (Aythya affinis) diving to depths of 1.2 and 2 m at thermoneutral (23 degrees C) and sub-thermoneutral (18 and 8 degrees C) temperatures. Birds dove while fasted and when feeding on blue mussels (Mytilus edulis). Substitution occurred if HIF or costs of diving above resting metabolic rate (RMR) were lower at 18 or 8 degrees C than at 23 degrees C, indicating reduction in the thermoregulatory part of RMR. For fasted scaup diving to 1.2 m, substitution from exercise heat was not apparent at either 18 or 8 degrees C. At 2 m depth, dive costs above RMR were reduced by 5% at 18 degrees C and by 40% at 8 degrees C, indicating substitution. At 1.2 m depth (with voluntary intake of only 14-17% of maintenance requirements), HIF did not differ between temperatures, indicating no substitution. However, at 2 m (intake 13-25% of maintenance), substitution from HIF was 23% of metabolizable energy intake at 18 degrees C and 22% at 8 degrees C. These results show that even with low HIF due to low intake rates, substitution from HIF can add to substitution from the heat of exercise. PMID- 16341523 TI - Safe cooling limits from exercise-induced hyperthermia. AB - We evaluated the cooling rate of hyperthermic subjects, as measured by three estimates of deep core temperatures (esophageal, rectal and aural canal temperatures), during immersion in a range of water temperatures. The objective of the study was to compare the three indices of core temperature and define safe cooling limits when using rectal temperature to avoid the development of hypothermia. On 4 separate days, seven subjects (four males, three females) exercised for 45.4+/-4.1 min at 65% V(O2)max at an ambient temperature of 39 degrees C, RH: 36.5%, until rectal temperature (T (re)) increased to 40.0 degrees C (39.5 degrees C for two subjects). Following exercise, the subjects were immersed in a circulated water bath controlled at 2, 8, 14 and 20 degrees C until T (re) returned to 37.5 degrees C. When T (re) reached normothermia during the cooling period (37.5+/-0.05 degrees C), both esophageal (T (es)) (35.6+/-1.3 degrees C) and aural canal (T (ac)) (35.9+/-0.9 degrees C) temperatures were approaching or reaching hypothermia, particularly during immersion in 2 degrees C water (T (es)=34.5+/-1.2 degrees C). On the basis of the heat loss data, the heat gained during the exercise was fully eliminated after 5.4+/-1.5, 7.9+/-2.9, 10.4+/-3.8 and 13.1+/-2.8 min of immersion in 2, 8, 14 and 20 degrees C water, respectively, with the coldest water showing a significantly faster cooling rate. During the immersion in 2 degrees C water, a decrease of only 1.5 degrees C in T (re) resulted in the elimination of 100% of the heat gained during exercise without causing hypothermia. This study would therefore support cooling the core temperature of hyperthermic subjects to a rectal temperature between 37.8 degrees C (during immersion in water >10 degrees C) and 38.6 degrees C (during immersion in water <10 degrees C) to eliminate the heat gained during exercise without causing hypothermia. PMID- 16341522 TI - Renal and endocrine changes in rats with inherited stress-induced arterial hypertension (ISIAH). AB - Hypertensive inbred rats (ISIAH; inherited stress-induced arterial hypertension) present with baseline hypertension (>170 mmHg in adult rats), but attain substantially higher values upon mild emotional stress. We aimed to characterize key parameters related to hypertension in ISIAH. Kidneys, adrenals, and systemic endocrine parameters were studied in ISIAH of different ages and compared to normotensive Wistar albino Glaxo (WAG) rats. Native organs were obtained for Western and PCR analysis. Perfusion-fixed organs were prepared for histopathology and quantitative histochemistry. Plasma renin and adrenal hormones were measured. Renal morphology was unaltered in ISIAH. The hypothalamic-pituitary adrenocortical (HPA) axis was constitutively upregulated with enlarged adrenal cortices and enhanced plasma corticosterone levels. Plasma renin activity was not different between groups, whereas aldosterone levels were in part reduced. Juxtaglomerular NO synthase type 1, cyclooxygenase type 2, and renin expression were significantly reduced, whereas tubular gene products related to sodium transport (bumetanide-sensitive Na, K, 2Cl cotransporter type 2; thiazide sensitive Na, Cl cotransporter; epithelial Na channel-alpha; 11beta hydroxysteroid dehydrogenase type 2) were increased. These data suggest enhanced volume conservation by the kidney. Our data define ISIAH as an attractive model for the renal components determining salt and water homeostasis in hypertension. The specific condition of a basally stimulated HPA axis is highlighted, including the option to study effects superimposed by emotional stress. PMID- 16341524 TI - Probing the limits of tool competence: experiments with two non-tool-using species (Cercopithecus aethiops and Saguinus oedipus). AB - Non-human animals vary in their ability to make and use tools. The goal of the present study was to further explore what, if anything, differs between tool users and non-tool-users, and whether these differences lie in the conceptual or motor domain. We tested two species that typically do not use tools-cotton top tamarins (Saguinus oedipus) and vervet monkeys (Cercopithecus aethiops)-on problems that mirrored those designed for prolific tool users such as chimpanzees. We trained subjects on a task in which they could choose one of two canes to obtain an out-of-reach food reward. After training, subjects received several variations on the original task, each designed to examine a specific conceptual aspect of the pulling problem previously studied in other tool-using species. Both species recognized that effective pulling tools must be made of rigid materials. Subsequent conditions revealed significant species differences, with vervets outperforming tamarins across many conditions. Vervets, but not tamarins, had some recognition of the relationship between a tool's orientation and the position of the food reward, the relationship between a tool's trajectory and the substance that it moves on, and that tools must be connected in order to work properly. These results provide further evidence that tool-use may derive from domain-general, rather than domain-specific cognitive capacities that evolved for tool use per se. PMID- 16341525 TI - Intrinsic joint kinematic planning. II: hand-path predictions based on a Listing's plane constraint. AB - This study was aimed at examining the assumption that three-dimensional (3D) hand movements follow specific paths that are dictated by the operation of a Listing's law constraint at the intrinsic joint level of the arm. A kinematic model was used to simulate hand paths during 3D point-to-point movements. The model was based on the assumption that the shoulder obeys a 2D Listing's constraint and that rotations are about fixed single-axes. The elbow rotations were assumed to relate linearly to those of the shoulder. Both joints were assumed to rotate without reversals, and to start and end rotating simultaneously with zero initial and final velocities. Model predictions were compared to experimental observations made on four right-handed individuals that moved toward virtual objects in "extended arm", "radial", and "frontal plane" movement types. The results showed that the model was partially successful in accounting for the observed behavior. Best hand-path predictions were obtained for extended arm movements followed by radial ones. Frontal plane movements resulted in the largest discrepancies between the predicted and the observed paths. During such movements, the upper arm rotation vectors did not obey Listing's law and this may explain the observed discrepancies. For other movement types, small deviations from the predicted paths were observed which could be explained by the fact that single-axis rotations were not followed even though the rotation vectors remained within Listing's plane. Dynamic factors associated with movement execution, which were not taken into account in our purely kinematic approach, could also explain some of these small discrepancies. In conclusion, a kinematic model based on Listing's law can describe an intrinsic joint strategy for the control of arm orientation during pointing and reaching movements, but only in conditions in which the movements closely obey the Listing's plane assumption. PMID- 16341526 TI - Intrinsic joint kinematic planning. I: reassessing the Listing's law constraint in the control of three-dimensional arm movements. AB - This study tested the validity of the assumption that intrinsic kinematic constraints, such as Listing's law, can account for the geometric features of three-dimensional arm movements. In principle, if the arm joints follow a Listing's constraint, the hand paths may be predicted. Four individuals performed 'extended arm', 'radial', 'frontal plane', and 'random mixed' movements to visual targets to test Listing's law assumption. Three-dimensional rotation vectors of the upper arm and forearm were calculated from three-dimensional marker data. Data fitting techniques were used to test Donders' and Listing's laws. The coefficient values obtained from fitting rotation vectors to the surfaces described by a second-order equation were analyzed. The results showed that the coefficients that represent curvature and twist of the surfaces were often not significantly different from zero, particularly not during randomly mixed and extended arm movements. These coefficients for forearm rotations were larger compared to those for the upper arm segment rotations. The mean thickness of the rotation surfaces ranged between approximately 1.7 degrees and 4.7 degrees for the rotation vectors of the upper arm segment and approximately 2.6 degrees and 7.5 degrees for those of the forearm. During frontal plane movements, forearm rotations showed large twist scores while upper arm segment rotations showed large curvatures, although the thickness of the surfaces remained low. The curvatures, but not the thicknesses of the surfaces, were larger for large versus small amplitude radial movements. In conclusion, when examining the surfaces obtained for the different movement types, the rotation vectors may lie within manifolds that are anywhere between curved or twisted manifolds. However, a two dimensional thick surface may roughly represent a global arm constraint. Our findings suggest that Listing's law is implemented for some types of arm movement, such as pointing to targets with the extended arm and during radial reaching movements. PMID- 16341527 TI - Effects of baclofen on the angular vestibulo-ocular reflex. AB - The purpose of this study was to determine the effect of baclofen, a GABA(B) agonist on the angular vestibulo-ocular reflex (aVOR). Model studies have shown that the aVOR comprises a "direct" pathway, which determines its high frequency gain g (1), and an indirect "velocity storage" pathway, which determines its low frequency characteristics. Velocity storage can be characterized by an integrator with a dominant time constant, T (VOR), and a gain g (0) that couples afferent information from the semicircular canals to the integrator. Baclofen preferentially shortens the velocity storage time constant in monkeys, but its effect on T (VOR), g (0), and g (1) in humans is unknown. Six subjects were tested after administration of a placebo or of 10, 20, or 30 mg of baclofen in a double-blind design. The aVOR was elicited in darkness with steps of rotation at 138 degrees /s, and g (1), g (0), and T (VOR) were determined from model fits of the slow phase velocity of the per- and post-rotatory nystagmus. Baclofen significantly reduced both T (VOR) and g (0) at dosages of 20 and 30 mg, but had no effect on g (1). Small reductions in g (0) were associated with large reductions in vestibular output. Thus, baclofen does not affect the direct aVOR pathway in humans, but controls the low frequency aVOR in two ways: it limits the input to velocity storage and modulates its time constant. We speculate that pre synaptic GABA(B) terminals in the vestibular nuclei are responsible for the control of the afferent input to velocity storage through g (0), while the post synaptic GABA(B) terminals are responsible for altering the duration of activity that reflects the time constant. The lack of effect of baclofen on the aVOR gain suggests that only GABA(A) receptors are utilized in the direct aVOR pathway. PMID- 16341528 TI - Imaging findings in primary intracranial atypical teratoid/rhabdoid tumors. AB - BACKGROUND: Intracranial atypical teratoid/rhabdoid tumors (AT/RT) are rare and extremely aggressive neoplasms seen primarily in childhood. Imaging features are often considered non-specific. However, correct diagnosis of AT/RT is important because these tumors have a markedly different clinical prognosis and require more aggressive therapy. OBJECTIVE: To determine the imaging features of AT/RT. MATERIALS AND METHODS: We retrospectively analyzed imaging findings in 11 patients with primary intracranial AT/RT presenting over a period of 5 years. CT (n=11), MR (n=7), clinical (n=11) and pathological (n=11) features were evaluated. FISH analysis showing monosomy of chromosome 22 (absence of bcr 22q11 locus) was available for three patients. Immunohistochemical staining for INI-1 (BAF47) was performed on all tumors. RESULTS: There were 11 patients, 6 boys and 5 girls. The age of presentation varied from 1 month to 15 years (average age 3 years 8 months). Six tumors were located in the posterior fossa and five in the supratentorial compartment. The tumors showed a hyperdense solid component (64%) that showed moderate to marked enhancement with contrast medium. On MR imaging, the predominant signal pattern was isointensity on T1-weighted images (57%) and T2 shortening with heterogeneity on T2-weighted images (86%). All tumors were large in size (average 4.2 x 3.7 cm), and there was a tendency for calcification (36%), hemorrhage (46%), necrosis (46%) and perifocal edema (100%). There was also a high tendency for subarachnoid dissemination, with five patients (46%) demonstrating brain and/or spinal metastasis. At follow-up (n=7), six patients showed local recurrence. At the time of recurrence, all these patients showed extensive leptomeningeal spread of the disease in both intracranial and intraspinal compartments. CONCLUSION: There are no specific imaging features for intracranial AT/RT. But a high tendency toward large size, a hyperdense solid component on CT scan with calcification, hemorrhage, necrosis and subarachnoid spread suggest that this tumor should be considered in the differential diagnosis of large pediatric intracranial tumors. PMID- 16341529 TI - MRI of the olfactory bulbs and sulci in human fetuses. AB - BACKGROUND: There is limited knowledge of the MRI pattern of the development of fetal olfactory bulbs and sulci. OBJECTIVE: To describe the MRI appearance of olfactory bulbs and sulci in normal in vivo fetuses according to gestational age. MATERIALS AND METHODS: Olfactory bulbs and sulci were retrospectively assessed on brain MRI examinations of 88 normal fetuses between 24 and 39 weeks gestational age. Two reference centres were involved in the study and both used routine protocols that included axial and coronal T2- and T1-weighted sequences at 1.5 T. The results were compared both with the commonly used neuropathological data in the literature and with personal neuropathological data. Pearson's chi-squared test or Fisher's exact test were performed. One case of olfactory agenesis associated with CHARGE syndrome was identified. RESULTS: T2-weighted coronal sequences were the most sensitive for detecting olfactory bulbs and sulci. Olfactory sulci were significantly better detected from 30 weeks onwards (90.9 100%; P<0.001). MRI showed a posteroanterior development of these sulci. Olfactory bulbs were better detected from 30 to 34 weeks (80-90.9%; P<0.002). Comparison with neuropathological data confirmed the posteroanterior development of the sulci and showed an important delay in detection of the olfactory structures (bulbs and sulci). No difference was observed between the two centres involved. CONCLUSIONS: To date, fetal MRI can depict olfactory sulci from 30 weeks gestational age onwards and olfactory bulbs from 30 to 34 weeks gestational age. This preliminary reference standard is useful to assess the normality of the olfactory system and to diagnose olfactory agenesis. PMID- 16341530 TI - Hydration of a glycoprotein: relative water affinity of peptide and glycan moieties. AB - Glycosylation, the most prevalent post-translational modification of proteins, affects a number of physical properties including the interactions with the surrounding aqueous solvent. Such glycan-water interactions have been discussed with respect to the increased solubility generally observed for glycoproteins, but experimental support of this correlation remains sparse. We have applied a two-channel calorimetric method to measure the free energy and enthalpy of hydration at 25 degrees C for the glycoprotein phytase (Phy) and a deglycosylated form (dgPhy) of the same protein. Comparisons of results for Phy and dgPhy show that the polypeptide moiety has a higher affinity for water than the glycans. In fact, at moderate hydration levels (approximately 0.3 g water/g macromolecule) the water uptake appears to be entirely governed by adsorption to the peptide groups. We conclude that strengthened interaction with the solvent is unlikely to be the mechanism underlying the increased solubility and lowered propensity of aggregation often reported to result from the glycosylation of proteins. PMID- 16341531 TI - The in vitro antitumor effect and in vivo tumor-specificity distribution of human mouse chimeric antibody against transferrin receptor. AB - Transferrin receptor (TfR/CD71) deserves attention as a selective target for cancer therapy due to its higher expression in tumors versus normal tissues. Also, it has been shown the mouse-derived monoclonal antibody against TfR can significantly inhibit the proliferation of tumor cells. Through constructing the chimeric antibody against TfR, the antigenicity of antibody can be weakened, and most importantly, the antitumor effect of antibody can be strengthened by the introduction of the human Fc fragment. In previous studies, we successfully constructed the human-mouse chimeric antibody against TfR (D2C) and demonstrated that its Fab fragment could specially recognize the TfR on the surface of target cells. In this study, through labeling the chimeric antibody D2C with 125I, we calculated the affinity constant (Ka) of 9.34-9.62x10(9) l/mol for this antibody according to the Scatchard drawing method. Moreover, in vivo studies in nude mice bearing human liver cancer (SMMC-7721) xenografts have shown that the radioactivity distribution ratio of 131I-D2C on T/NT was 2-14:1 or 3-21:1 on the seventh day after intraperitoneal or intratumoral injection of 131I-labeled D2C (131I-D2C). These evidences indicated that the in vivo distribution of D2C display the characteristics of certain tumor-specificity localization. In vitro studies, D2C can induce the apoptosis of K562 through the mitochondria death pathway and arrest the cell at G1 phase, as determined by cell cycle analysis. Using the human tumor cells (K562, CEM, and SMMC-7721) expressing TfR as target cells, and normal human PBMC as effector cells, Fc fragment of D2C can perform both the antibody-dependent cell-mediated cytotoxicity and the complement dependent cytotoxicity. Together, it was demonstrated that the D2C display a tumor-specificity distribution, and has a strong antitumor effect. Thus, it has the potential therapeutic significance. PMID- 16341532 TI - Preferential extravasation and accumulation of liposomal vincristine in tumor comparing to normal tissue enhances antitumor activity. AB - To quantitatively evaluate the extravasation, accumulation and selectivity to tumor tissues of liposomal vincristine (LV), dorsal skin-fold window chambers on athymic mice with or without LX-1, a human small cell lung cancer, xenograft implants and fluorescent intravital microscopy imaging were used. In vitro studies show that minimal loss of fluorescence marker DiI from liposomes occurs after 4 days of inoculation in murine plasma, and the release profiles of DiI-LV and LV were essentially the same with approximately 40% of the encapsulated vincristine sulfate (VCR) released after 26 h. Significantly faster extravasation of DiI-LV from tumor vessels was shown compared to non-tumor tissue after single dose i.v. administration. The relative interstitial amounts at 60 min (RIA(60)) for tumor and non-tumor tissues were 0.837+/-0.314 and 0.012+/-0.091, respectively (P=0.01). DiI-LV accumulation was significantly higher in tumor than in normal tissue, which continued beyond 48 h. Both DiI-LV and LV showed significant antitumor effects in window chambers and in flank tumors, compared with controls and VLS alone. The preferential extravasation of DiI-LV from tumor vasculature as well as its differential retention in tumor tissue provides the basis for the enhancement in antitumor activity of LV over VCR. PMID- 16341533 TI - Sexual activity, sexual dysfunction and associated help-seeking behaviours in middle-aged and older adults in Spain: a population survey. AB - To study sexual activity, the prevalence of sexual dysfunction and related help seeking behaviours among mature adults in Spain, a telephone survey was conducted in Spain in 2001-2002. This was completed by 750 men and 750 women aged 40-80 years. Eighty-eight percent of men and 66% of women had engaged in sexual intercourse during the 12 months preceding the interview. Early ejaculation (31%) and lack of sexual interest (17%) were the most common male sexual problems. A lack of sexual interest (36%) and an inability to reach orgasm (28%) were the most common female sexual problems. Approximately 80% of men and women with a sexual problem had not sought help from a health professional. Many men and women in Spain report continued sexual interest and activity into middle age and beyond. Although a number of sexual problems are highly prevalent, few people seek medical help. PMID- 16341534 TI - Revision of prolapsed feeding gastrostomy with a modified Janeway 'gastric tube'. AB - After creation of open or percutaneous endoscopic gastrostomy, gastric prolapse and leak of gastric contents may cause serious skin rash and infection which are often difficult to treat. We present four patients in whom these problems were solved with gastrostomy revision by a modified Janeway 'gastric tube' technique. The patients were aged 7 months and 7, 10 and 16 years at the time of the revision. The underlying conditions were hypoxic encephalopathy with epilepsy, infantile spasm and epilepsy with arthrogryposis, dystonic tetraplegy, and total colon aganglionosis. All patients had gastrostomy prolapse with peristomal skin rash and cellulitis. Prior to modified Janeway revision, the four patients had undergone a total of 16 failed attempts to cure the prolapse. At the operation, the previous gastrostomy was detached and closed. A longitudinal gastric tube of 6 cm was created along the greater curvature with a GIA stapler and brought through the abdominal wall leaving 3-5 cm of free intra-abdominal gastric tube. A balloon catheter was left for 6 weeks, and replaced with a long Mickey tube according to patient's or caretaker's preference. There were no surgical complications. Hospitalisation after revision was median 6 (range 4-11) days. Six weeks after the revision, prolapse, leak and peristomal infections were cured in all patients, and feeding through the gastrostomy presented no problems. One patient underwent minor excision of excess stomal mucosa. Two patients opted for Mickey tube, two for a feeding catheter. A median of 9 (6-16) months after the revision, all patients have a functioning gastrostomy without prolapse or leak. Modified Janeway 'gastric tube' revision is feasible and, within short to medium term follow-up, controls efficiently gastrostomy prolapse and leak. PMID- 16341535 TI - Diagnostic laparoscopy-assisted cholangiography in infants with prolonged jaundice. AB - Cholangiography is often crucial for establishing the definitive cause of neonatal jaundice. We present our protocol for using laparoscopy-assisted cholangiography in infants with prolonged jaundice and discuss its benefits. Firstly, a 5 mm supra-umbilical trocar is introduced to create a port for a 0 degrees laparoscope. A 5 mm trocar is then inserted through a right subcostal incision to allow the liver and gallbladder to be visualized. If the gallbladder is of good size, the fundus is exteriorized through the right subcostal trocar site and a catheter is inserted into the gallbladder for cholangiography. If the gallbladder is atretic, the fundus is not exteriorized and a laparotomy is performed for open intraoperative cholangiography because the lumen of an atretic gallbladder is usually not fully patent and cholangiography through its exteriorized fundus often fails. We reviewed 18 jaundiced infants thought to have biliary atresia (BA) who had laparoscopy-assisted cholangiography. At laparoscopy, four patients had good sized gallbladders and minimal to mild liver fibrosis. They underwent cholangiography via the exteriorized fundus, and BA in two cases and biliary hypoplasia in two cases were identified. The remaining 14 had atretic gallbladders and varying degrees of liver fibrosis. Cholangiography via the exteriorized fundus was performed in one patient, but failed and converted to open cholangiography. Open intraoperative cholangiography identified BA in all 14 cases. All BA cases progressed to Kasai portoenterostomy directly after diagnosis. Laparoscopy is used to determine the type of cholangiography to be performed based on the appearance of the gallbladder and this simple, accurate, and safe protocol allows the anatomical structure of the biliary tree to be obtained accurately with minimal surgical intervention. PMID- 16341536 TI - Serial tightening of Prolene mesh in the repair of a large ventral hernia. AB - A 9-month-old female baby was brought to our hospital with a large ventral hernia which had developed after conservative treatment of an exomphalos. A hitherto undescribed technique involving serial tightening of a Prolene mesh was utilised to close the ventral hernia. We were able to achieve a good result within a short period of 2 weeks, without resorting to ventilation. We propose this procedure as an alternative to other existing techniques in similar situations. PMID- 16341537 TI - The dimensional accuracy for preparation of the femoral cavity in HIP arthroplasty. A comparison between manual- and robot-assisted implantation of hip endoprosthesis stems in cadaver femurs. AB - INTRODUCTION: The aim of the study was to determine the precision of the preparation of the femoral cavity in cementless hip arthroplasty. We compared the bone-prosthesis interface after manual- and robot-assisted implantation of the stems. MATERIAL AND METHODS: After plastination the specimens were cut to slices of 2.5 mm and documented by microradiography. The interface between prosthesis and bone was measured digitally with a specially designed software. RESULTS: The manually implanted prostheses showed an average full-contact area of 60% and an average area of gaps of 40% with an average height of 0.8 mm. The robot-assisted implanted stems had a significantly higher bone-prosthesis interface area of 93% and gap areas of less than 0.2 mm depth. Destructions of the spongeous bone were seen with the manually implanted stems but not after robot-assisted implantations. There was one stem fracture during the manual preparation of a stem. Microradiographic examination showed microfractures in the region of the lesser trochanter in two femora after robot-assisted implantation CONCLUSION: The robot-assisted technique highly increased the fitting area at the bone-prosthesis interface. The main reason for the fractures might be the absence of a robot done marker for the ideal implantation height with the applied type of prosthesis. PMID- 16341538 TI - Arthroscopic reconstruction of the acromioclavicular joint disruption: surgical technique and preliminary results. AB - INTRODUCTION: Numerous operative procedures have been described for the reconstruction of acromioclavicular joint separation; however, the arthroscopic reconstruction has been rarely reported. Therefore, our objective was to propose a new technique of arthroscopic acromioclavicular joint surgery and to evaluate the preliminary results. MATERIALS AND METHODS: Thirteen patients with a mean follow-up of 18 months underwent the arthroscopic acromioclavicular joint reconstruction using suture anchors and small titanium plate. The average age was 40.2 years (range 23-54 years). The shoulders were evaluated using Constant score and radiographs. The indications for surgery included acromioclavicular joint dislocation Rockwood type IV-V. RESULTS: Twelve patients returned to their work without pain within 3 months after operation. The average Constant score at last follow-up was 95. Postoperative radiographs confirmed anatomic reduction in ten patients, residual subluxation in two patients and redislocation of the joint in one patient. One patient had radiographic evidence of coracoclavicular ossification. All patients but one were satisfied with results and cosmetic appearance. CONCLUSION: Considering its less morbidity, excellent cosmesis, no need of hardware removal, and minimal complications from breakage or migration of metal implants, this new technique offers an attractive alternative in acromioclavicular joint stabilization. PMID- 16341539 TI - Disseminated peritoneal leiomyomatosis coexisting with leiomyoma of the uterine body. AB - The authors present an extremely rare case of disseminated peritoneal leiomyomatosis (DPL) coexisting with leiomyoma of the uterine body in a 32-year old woman. The patient had undergone a caesarean section 12 years ago and used oral contraceptives since then. Before the present surgery leiomyoma of the left corner of the uterine body was diagnosed. DPL was found on the uterine serous membrane, Douglas's cavity, vesicouterine recess, the great omentum and abdominal peritoneum. There were no DPL nodules on the surface of the liver, kidneys and bowels. Saving operation was performed and only leiomyoma and the great omentum were resected. Immunohistochemical analysis using smooth muscle actin and HHF-35 antibodies showed the same reactivity of leiomyoma and DPL cells and proved the intra-operative diagnosis. Our case corroborates that DPL may occur in women of reproductive age and in patients who use oral contraceptives over a long span of time. That both leiomyoma and DPL are hormonally dependent could also be proved. PMID- 16341540 TI - One-year outcomes after photodynamic therapy in patients with age-related macular degeneration with poor baseline visual acuity. AB - BACKGROUND: Photodynamic therapy with verteporfin (PDT) has been demonstrated in randomized controlled trials to be a safe and effective therapy for choroidal neovascular membranes (CNV) secondary to age-related macular degeneration (AMD). Limited information is available on the prognosis with PDT for patients who fell outside the inclusion criteria for the clinical trials, however. The purpose of this study is to describe the clinical course of patients with CNV lesions in AMD treated with PDT, with baseline visual acuities sufficiently poor to warrant exclusion from previous randomized trials. METHODS: Retrospective case series. Ten consecutive patients with CNV secondary to exudative AMD treated with PDT with baseline visual acuity less than 34 ETDRS letters were followed for 1 year. The main outcome was median change in visual acuity on the ETDRS chart. RESULTS: The median change in acuity over 12 months was + 13 letters. All patients lost <3 lines of ETDRS acuity, and eight of ten patients (80%) gained at least one line of vision, over 12 months. CONCLUSIONS: In our series, patients with low visual acuity at baseline appeared to respond to PDT on both visual acuity and fluorescein angiographic measurements. PDT treatment may be considered for selected patients with these baseline characteristics. PMID- 16341541 TI - Effect of additive preoperative latanoprost treatment on the outcome of filtration surgery. AB - BACKGROUND: Long-term glaucoma medication has been suspected to be a risk factor for bleb failure following trabeculectomy. It was the goal of our study to investigate whether additive preoperative treatment with latanoprost has an effect on the outcome of subsequent trabeculectomy. METHODS: We retrospectively analysed the outcome of trabeculectomy in 32 eyes of 16 patients who had received bilateral trabeculectomy within the Mainz-II study. This study had been designed to examine the effect of 6 months' use of topical latanoprost therapy on iris darkening. The first eye of each patient was operated upon without preoperative latanoprost therapy; the other eye was operated on after 6 months' treatment with latanoprost in addition to existing medical glaucoma treatment. We analysed the outcome of these patients and compared success rates and average intra-ocular pressures (IOPs) between the latanoprost-treated and the untreated partner eyes. Success was defined as an average postoperative IOP of 18 mmHg or less. RESULTS: We obtained IOP values from 13 of 16 patients, with a mean follow-up period of 6.8 years. Three patients were lost to follow-up. In both groups eight eyes (61.5%) had an average postoperative IOP (average of all IOPs measured in the follow-up period) of 18 mmHg or less. The mean average postoperative IOPs in the control group and in the latanoprost group were 15.2 mmHg and 15.8 mmHg, respectively. Intra-individual comparison revealed that one pair of eyes had equal IOPs, while six eyes in each group showed better postoperative IOP control than the respective partner eye. However, mean delta IOPs (maximum preoperative IOP minus average postoperative IOP) were 17.8 mmHg and 14.2 mmHg for the control group and latanoprost group, respectively. CONCLUSION: In this small group 6 months of additive preoperative treatment with latanoprost did not have a statistically significant effect on the success rate of trabeculectomy or on the postoperative IOP level following trabeculectomy. However, trabeculectomy in eyes preoperatively receiving latanoprost for 6 months might lead to a slightly smaller delta IOP than in eyes naive to prostaglandins. PMID- 16341542 TI - Julius Bernstein (1839-1917): pioneer neurobiologist and biophysicist. AB - Julius Bernstein belonged to the Berlin school of "organic physicists" who played a prominent role in creating modern physiology and biophysics during the second half of the nineteenth century. He trained under du Bois-Reymond in Berlin, worked with von Helmholtz in Heidelberg, and finally became Professor of Physiology at the University of Halle. Nowadays his name is primarily associated with two discoveries: (1) The first accurate description of the action potential in 1868. He developed a new instrument, a differential rheotome (= current slicer) that allowed him to resolve the exact time course of electrical activity in nerve and muscle and to measure its conduction velocity. (2) His 'Membrane Theory of Electrical Potentials' in biological cells and tissues. This theory, published by Bernstein in 1902, provided the first plausible physico-chemical model of bioelectric events; its fundamental concepts remain valid to this day. Bernstein pursued an intense and long-range program of research in which he achieved a new level of precision and refinement by formulating quantitative theories supported by exact measurements. The innovative design and application of his electromechanical instruments were milestones in the development of biomedical engineering techniques. His seminal work prepared the ground for hypotheses and experiments on the conduction of the nervous impulse and ultimately the transmission of information in the nervous system. Shortly after his retirement, Bernstein (1912) summarized his electrophysiological work and extended his theoretical concepts in a book Elektrobiologie that became a classic in its field. The Bernstein Centers for Computational Neuroscience recently established at several universities in Germany were named to honor the person and his work. PMID- 16341543 TI - Do cortical maps depend on the timing of sensory input? Experimental evidence and computational model. AB - Fast adaptations in the functional organization of primary sensory cortex are generally assumed to result from changes of network connectivity. However, the effects of intrinsic neuronal excitability alterations due to the activation of neighboring cortical representational zones, which might as well account for the changes of cortical representative maps, have been paid little attention to. In a recent experiment (Braun et al. 2000b) we showed by neuromagnetic source imaging that random or fixed sequence stimulation of three digits of both hands led to stimulation-timing-induced changes in primary somatosensory (SI) cortical maps. The distance between the cortical representation of thumb and middle finger became significantly shorter during the fixed sequence stimulation. The analysis on the time course of the cortical map changes revealed that these reorganizations occurred within minutes and were fully reversible. The previously reported results were interpreted as the involvement of a superordinate center responsible for detecting and activating the appropriate maps. Here we present an alternative parsimonious explanation that is supported by a computational model. Based on the experimental evidence, we developed a simple model that took intrinsic neuronal excitability together with subthreshold activation into account and assumed partial cortical overlap of the representational zones of neighboring digits. Furthermore, in the model the neuronal excitability decayed slowly with respect to the stimulation frequency. The observed cortical map changes in the experiment could be reproduced by the two-layer feed-forward computational network. Our model thus suggests that the dynamic shifts of cortical maps can be explained by the state and time course of intrinsic neuronal excitability and subthreshold activation, without involving changes in network connectivity. PMID- 16341545 TI - Feature integration across perception and action: event files affect response choice. AB - Five experiments investigated the spontaneous integration of stimulus and response features. Participants performed simple, prepared responses (R1) to the mere presence of Go signals (S1) before carrying out another, freely chosen response (R2) to another stimulus (S2), the main question being whether the likelihood of repeating a response depends on whether or not the stimulus, or some of its features, are repeated. Indeed, participants were more likely to repeat the previous response if stimulus form or color was repeated than if it was alternated. The same was true for stimulus location, but only if location was made task-relevant, whether by defining the response set in terms of location, by requiring the report of S2 location, or by having S1 to be selected against a distractor. These findings suggest that task-relevant stimulus and response features are spontaneously integrated into independent, local event files, each linking one stimulus to one response feature. Upon reactivation of one member of the binary link activation is spread to the other, thereby increasing the likelihood to repeat a response if one or more stimulus features are repeated. These findings support the idea that both perceptual events and action plans are cognitively represented in terms of their features, and that feature-integration processes cross borders between perception and action. PMID- 16341544 TI - Nitric oxide scavenging by barley hemoglobin is facilitated by a monodehydroascorbate reductase-mediated ascorbate reduction of methemoglobin. AB - NADH-dependent NO scavenging in barley extracts is linked to hemoglobin (Hb) expression and is inhibited by SH-reagents. Barley Hb has a single cysteine residue. To determine whether this cysteine was critical for NO scavenging, barley Hb and a mutated version, in which the single Cys(79) was replaced by Ser, were over-expressed in Escherichia coli and purified to near homogeneity. The purified proteins exhibited very low NO-scavenging activity (12-14 nmol min(-1) mg(-1) protein) in the presence of NADH or NADPH. This activity was insensitive to SH-reagents. Addition of an extract from barley roots to either of the purified proteins resulted in high NADH-dependent NO turnover in a reaction that was sensitive to SH-reagents. A protein was purified from barley roots and identified by mass-spectrometry analysis as a cytosolic monodehydroascorbate reductase. It efficiently supported NADH-dependent NO scavenging in the presence of either native or mutated barley Hb. Ascorbate strongly facilitated the rate of metHb reduction. The K (m) for Hb was 0.3 microM, for ascorbate 0.6 mM and for NADH 4 microM. The reaction in the presence of monodehydroascorbate reductase was sensitive to SH-reagents with either form of the Hb. We conclude that metHb reduction and NO turnover do not involve direct participation of the Cys(79) residue of barley Hb. NO scavenging is facilitated by monodehydroascorbate reductase mediating a coupled reaction involving ferric Hb reduction in the presence of ascorbate and NADH. PMID- 16341546 TI - Spreading the sparing: against a limited-capacity account of the attentional blink. AB - The identification of the second of two targets presented in close succession is often impaired--a phenomenon referred to as the attentional blink. Extending earlier work (Di Lollo, Kawahara, Ghorashi, and Enns, in Psychological Research 69:191-200, 2005), the present study shows that increasing the number of targets in the stream can lead to remarkable improvements as long as there are no intervening distractors. In addition, items may even recover from an already induced blink whenever they are preceded by another target. It is shown that limited memory resources contribute to overall performance, but independent of the attentional blink. The findings argue against a limited-capacity account of the blink and suggest a strong role for attentional control processes that may be overzealously applied. PMID- 16341547 TI - Thyroid and pituitary gland development from hatching through metamorphosis of a teleost flatfish, the Atlantic halibut. AB - Fish larval development, not least the spectacular process of flatfish metamorphosis, appears to be under complex endocrine control, many aspects of which are still not fully elucidated. In order to obtain data on the functional development of two major endocrine glands, the pituitary and the thyroid, during flatfish metamorphosis, histology, immunohistochemistry and in situ hybridization techniques were applied on larvae of the Atlantic halibut (Hippoglossus hippoglossus), a large, marine flatfish species, from hatching through metamorphosis. The material was obtained from a commercial hatchery. Larval age is defined as day-degrees (D degrees =accumulated daily temperature from hatching). Sporadic thyroid follicles are first detected in larvae at 142 D degrees (27 days post-hatch), prior to the completion of yolk sack absorption. Both the number and activity of the follicles increase markedly after yolk sack absorption and continue to do so during subsequent development. The larval triiodothyronine (T(3)) and thyroxine (T(4)) content increases, subsequent to yolk absorption, and coincides with the proliferation of thyroid follicles. A second increase of both T(3) and T(4) occurs around the start of metamorphosis and the T(3) content further increases at the metamorphic climax. Overall, the T(3) content is lower than T(4). The pituitary gland can first be distinguished as a separate organ at the yolk sack stage. During subsequent development, the gland becomes more elongated and differentiates into neurohypophysis (NH), pars distalis (PD) and pars intermedia (PI). The first sporadic endocrine pituitary cells are observed at the yolk sack stage, somatotrophs (growth hormone producing cells) and somatolactotrophs (somatolactin producing cells) are first observed at 121 D degrees (23 days post-hatch), and lactotrophs (prolactin producing cells) at 134 D degrees (25 days post-hatch). Scarce thyrotrophs are evident after detection of the first thyroid follicles (142 D degrees ), but coincident with a phase in which follicle number and activity increase (260 D degrees ). The somatotrophs are clustered in the medium ventral region of the PD, lactotrophs in the anterior part of the PD and somatolactotrophs are scattered in the mid and posterior region of the pituitary. At around 600 D degrees , coinciding with the start of metamorphosis, somatolactotrophs are restricted to the interdigitating tissue of the NH. During larval development, the pituitary endocrine cells become more numerous. The present data on thyroid development support the notion that thyroid hormones may play a significant role in Atlantic halibut metamorphosis. The time of appearance and the subsequent proliferation of pituitary somatotrophs, lactotrophs, somatolactotrophs and thyrotrophs indicate at which stages of larval development and metamorphosis these endocrine cells may start to play active regulatory roles. PMID- 16341548 TI - Catfish CC chemokines: genomic clustering, duplications, and expression after bacterial infection with Edwardsiella ictaluri. AB - Chemokines are a family of structurally related chemotactic cytokines that regulate the migration of leukocytes, under both physiological and inflammatory conditions. CC chemokines represent the largest subfamily of chemokines with 28 genes in mammals. Sequence conservation of chemokines between teleost fish and higher vertebrates is low and duplication and divergence may have occurred at a significantly faster rate than in other genes. One feature of CC chemokine genes known to be conserved is genomic clustering. CC chemokines are highly clustered within the genomes of human, mouse, and chicken. To exploit knowledge from comparative genome analysis between catfish and higher vertebrates, here we mapped to bacterial artificial chromosome (BAC) clones 26 previously identified catfish (Ictalurus sp.) chemokine cDNAs. Through a combination of hybridization and fluorescent fingerprinting, 18 fingerprinted contigs were assembled from BACs containing catfish CC chemokine genes. The catfish CC chemokine genes were found to be not only highly clustered in the catfish genome, but also extensively duplicated at various levels. Comparisons of the syntenic relationships of CC chemokines may help to explain the modes of duplication and divergence that resulted in the present repertoire of vertebrate CC chemokines. Here we have also analyzed the expression of the transcripts of the 26 catfish CC chemokines in head kidney and spleen in response to bacterial infection of Edwardsiella ictaluri, an economically devastating catfish pathogen. Such information should pinpoint research efforts on the CC chemokines most likely involved in inflammatory responses. PMID- 16341549 TI - Towards compendia of negative genetic association studies: an example for Alzheimer disease. AB - Most genetic sequence variants that contribute to variability in complex human traits will have small effects that are not readily detectable with population samples typically used in genetic association studies. A potentially valuable tool in the gene discovery process is meta-analysis of the accumulated published data, but in order to be valid these require a sample of studies representative of the true genetic effect and thus hypothetically should include some positive and an abundance of negative reports. A survey of the literature on association studies for Alzheimer disease (AD) from January 2004-April 2005, identified 138 studies, 86 of which reported positive findings other than for apolipoprotein E (APOE), strongly indicative of publication bias. We report here an analysis of 62 genetic markers, tested for association with AD risk as well as for possible effects upon quantitative indices of AD severity (mini-mental state examination scores, age-at-onset, and cerebrospinal fluid (CSF) beta-amyloid (Abeta) and CSF tau proteins). Within this set, only modest signals were present that, with the exception of APOE are easily lost when corrections for multiple hypotheses are applied. In isolation, results are thus broadly negative. Genes studied encompass both novel candidates as well as several recently claimed to be associated with AD (e.g. urokinase plasminogen activator (PLAU) and acetyl-coenzyme A acetyltransferase 1 (ACAT1)). By reporting these data we hope to encourage the publication of gene compendia to guide further studies and aid future meta analyses aimed at resolving the involvement of genes in complex human traits. PMID- 16341550 TI - Aberrant splicing in MLH1 and MSH2 due to exonic and intronic variants. AB - Single base substitutions in DNA mismatch repair genes which are predicted to lead either to missense or silent mutations, or to intronic variants outside the highly conserved splicing region are often found in hereditary nonpolyposis colorectal cancer (HNPCC) families. In order to use the variants for predictive testing in persons at risk, their pathogenicity has to be evaluated. There is growing evidence that some substitutions have a detrimental influence on splicing. We examined 19 unclassified variants (UVs) detected in MSH2 or MLH1 genes in patients suspected of HNPCC for expression at RNA level. We demonstrate that 10 of the 19 UVs analyzed affect splicing. For example, the substitution MLH1,c.2103G > C in the last position of exon 18 does not result in a missense mutation as theoretically predicted (p.Gln701His), but leads to a complete loss of exon 18. The substitution MLH1,c.1038G > C (predicted effect p.Gln346His) leads to complete inactivation of the mutant allele by skipping of exons 10 and 11, and by activation of a cryptic intronic splice site. Similarly, the intronic variant MLH1,c.306+2dupT results in loss of exon 3 and a frameshift mutation due to a new splice donor site 5 bp upstream. Furthermore, we confirmed complete exon skipping for the mutations MLH1,c.1731G > A and MLH1,c.677G > A. Partial exon skipping was demonstrated for the mutations MSH2,c.1275A > G, MLH1,c.588+5G > A, MLH1,c.790+4A > G and MLH1,c.1984A > C. In contrast, five missense mutations (MSH2,c.4G > A, MSH2,c.2123T > A, MLH1,c.464T > G, MLH1,c.875T > C and MLH1,c.2210A > T) were found in similar proportions in the mRNA as in the genomic DNA. We conclude that the mRNA examination should precede functional tests at protein level. PMID- 16341551 TI - Expression and localisation of alphav integrins in human odontoblasts. AB - Integrin alphabeta heterodimers mediate adhesion to the extracellular matrix and at cell-cell contacts and initiate intracellular signalling cascades in response to a variety of inductive factors. Apart from the expression of alphavbeta3 that we have previously reported, little is known about the expression of integrins in odontoblasts. Here, we investigated the expression of alphav-binding beta integrin subunits in healthy human dental pulp in vivo and in odontoblasts differentiated in vitro. Reverse transcription/polymerase chain reaction analysis revealed the expression of alphav, beta1, beta5 and beta8 integrin mRNA, but not beta6, in whole pulp cells. Flow cytometry showed that the alphav and beta1 subunits were the most intensely expressed. Immunohistochemistry demonstrated that the beta1 subunit was localised in newly differentiated odontoblasts in the root and in mature odontoblasts in the crown, including their intradentinal cell processes. The alphav chain was predominantly expressed by mature odontoblasts and alphavbeta5 was only observed in mature odontoblasts. In vitro differentiated odontoblasts expressed genes for alphav, beta1 and beta5, but not for beta6 and beta8. A comparison of integrin profiles between cultured pulp cells and in vitro differentiated odontoblasts revealed that odontoblast maturation was characterised by a significant increase in the expression of alphav and beta1 subunits and alphavbeta5 integrin. The beta8 subunit was detected in nerve cells only. Histological analysis of teeth from alphav knockout mice showed no obvious structural modification in the odontoblast layer. Thus, human mature odontoblasts express alphavbeta3, alphavbeta5 and perhaps alphavbeta1 integrins, with the possible presence of alpha-beta1 pairs. The roles that these molecules play in the exchange of information throughout the odontoblast layer remain to be determined. PMID- 16341552 TI - Females make tough neighbors: sex-specific competitive effects in seedlings of a dioecious grass. AB - If males and females of a species differ in their effect on intraspecific competition then this can have significant ecological and evolutionary consequences because it can lead to size and mortality disparities between the sexes, and thus cause biased population sex ratios. If the degree of sexual dimorphism of competitive effect varies across environments then this variation can generate sex ratio variation within and between populations. In a California population of Distichlis spicata, a dioecious grass species exhibiting extreme within-population sex ratio variation (spatial segregation of the sexes), I evaluated the intraspecific competitive effects of male and female D. spicata seedlings in three soil types. The sex of seedlings was determined using a RAPD PCR marker co-segregating with female phenotype. Distichlis spicata seedlings, regardless of sex, were six times larger when grown with male versus female conspecific seedlings in soil from microsites where the majority of D. spicata plants are female, and this sexual dimorphism of competitive effect was weaker or did not occur in other soil types. This study suggests that it is not just the higher costs of female versus male reproduction itself that cause spatial segregation of the sexes in D. spicata, but that differences in competitive abilities between the sexes--which occur as early as the seedling stage--can generate sex ratio variation. PMID- 16341553 TI - Density-dependent responses of fawn cohort body mass in two contrasting roe deer populations. AB - We investigated the influence of population density on juvenile body mass in two contrasting roe deer populations, in Sweden (Bogesund) and France (Chize), in which density was monitored for >or=15 years. We investigated the effect of population density and climatic conditions on cohort performance. We predicted that: (1) body mass of growing fawns should be sensitive to environmental changes, showing marked between-year variation (i.e., cohort effects), (2) fawns in the less productive (weakly seasonal, weakly predictable summer weather) habitat of Chize should show stronger density-dependent responses due to more severe food competition during summer than fawns in the more productive (markedly seasonal, moderately predictable summer weather) habitat of Bogesund, and (3) fawns at Bogesund should be heavier both in absolute terms and relative to their size than their conspecifics in Chize due to a higher degree of fat accumulation in northern environments. In both study sites we found marked cohort variation and clear effects of density, with body mass varying by as much as 29% over years. While neither summer nor winter climate influenced fawn body mass at Bogesund, fawns tended to be lighter after summers with high temperatures at Chize. In addition, fawns were heavier after acorn mast years experienced in utero at Bogesund. As expected, the strength of the density-dependent response of fawn body mass was greater at Chize than at Bogesund. For a given density, male fawns were consistently heavier than females in both sites. Lastly, both sexes at Bogesund had higher absolute body mass and were larger for a given body size than in Chize. Our results clearly demonstrate that absolute density is a poor predictor of roe deer performance and supports the view that habitat quality has an overwhelming importance for determining fawn body mass in roe deer populations. PMID- 16341554 TI - Purification of nattokinase by reverse micelles extraction from fermentation broth: effect of temperature and phase volume ratio. AB - Nattokinase is a novel fibrinolytic enzyme that is considered to be a promising agent for thrombosis therapy. In this study, reverse micelles extraction was applied to purify and concentrate nattokinase from fermentation broth. The effects of temperature and phase volume ratio used for the forward and backward extraction on the extraction process were examined. The optimal temperature for forward and backward extraction were 25 degrees C and 35 degrees C respectively. Nattokinase became more thermosensitive during reverse micelles extraction. And it could be enriched in the stripping phase eight times during backward extraction. It was found that nattokinase could be purified by AOT reverse micelles with up to 80% activity recovery and with a purification factor of 3.9. PMID- 16341555 TI - Penetrating spinal injury with wooden fragments causing cauda equina syndrome: case report and literature review. AB - STUDY DESIGN: Case report. OBJECTIVE: To report an unusual case of cauda equina syndrome following penetrating injury to the lumbar spine by wooden fragments and to stress the importance of early magnetic resonance imaging (MRI) in similar cases. SUMMARY OF BACKGROUND DATA: A 22-year-old girl accidentally landed on wooden bannister and sustained a laceration to her back. She complained of back pain but had fully intact neurological function. The laceration in her back was explored and four large wooden pieces were removed. However 72 h later, she developed cauda equina syndrome. MRI demonstrated the presence of a foreign body between second and third lumbar spinal levels following which she underwent emergency decompressive laminectomy and the removal of the multiple wooden fragments that had penetrated the dura. RESULTS: Post-operatively motor function in her lower limbs returned to normal but she continued to require a catheter for incontinence. At review 6 months later, she was mobilising independently but the incontinence remained unchanged. CONCLUSION: There are no reported cases in the literature of wooden fragments penetrating the dura from the back with or without the progression to cauda equina syndrome. The need for a high degree of suspicion and an early MRI scan to localise any embedded wooden fragments that may be separate from the site of laceration is emphasized even if initial neurology is intact. PMID- 16341556 TI - Distribution patterns of transforaminal injections in the cervical spine evaluated by multi-slice computed tomography. AB - Transforaminal injections are sometimes used for the diagnosis and treatment of painful conditions in the lumbar and to a lesser degree in the cervical spine. The technique is most often used when investigating/treating radiculopathy caused by degenerative disease. But how selective are the nerve root blocks? What possible structures other than the intended nerve root are affected from such injections? This study was undertaken in order to try to answer these questions, as no study focusing on the possible spread from the transforaminal selective nerve root blocks in the cervical spine has been performed earlier. In three groups of patients, each group including three patients, we injected three different volumes (0.6, 1.1 and 1.7 ml) with a transforaminal technique in the cervical spine. In all the injections, a small amount of contrast media was added. The spread of the injections were then investigated using multi-slice computed tomography with reconstructions. The imaging revealed a possible effect on other nerve roots than the intended ones when a larger volume was used for the root blocks. The spread was related to the injected volume as well as to local anatomy (size of foraminal area). In this study, only 0.6-ml injections could be accepted for being selective enough for diagnostic investigations. PMID- 16341566 TI - Development of a total colonoscopy rat model with endoscopic submucosal injection of the cecal wall. AB - BACKGROUND: Experimental models of colorectal tumor require either laparotomy for induction or anastomosis following resection. The long murine cecum avoids the need for an anastomosis, making the cecum the preferred site for induction. This study aimed to evaluate total colonoscopy with submucosal injection of cecal wall (TCWI) in rats in terms of failure rate (FR), complication rate (CR), and reproducibility (R). METHODS: A bolus of bowel prep was given. Anesthesia was injected intraperitoneally. A video fiberscope (5.9 mm outer diameter, 180/90 degrees up/down bending, 100/100 degrees right/left bending, 103 cm working length, 120 degrees view field, and 2.0 mm channel) allowed for irrigation and suction. Saline 1 ml was injected in the cecal wall through a 4-mm-long, 23-gauge needle placed on a 3-mm wire, resulting in a blister. FR was a failure to reach and inject the cecum. Rats were allowed to recover. CR was measured at necropsy. R was assessed by comparing TCWI time, FR, and CR for three investigators. Sample size of 120 (type I error, 0.05; power, 80%) was based on a pilot study. Data are presented as median (range). RESULTS: A total of two of 122 rats (1.6%) died after prep or anesthesia. Bowel prep resulted in 99.1% evacuation of solid feces. A total of 120 male Sprague-Dawley retired breeders weighing 592 g (range, 349 780) underwent TCWI. Scope depth was 28 cm (range, 20-36). Irrigating fluid was 290 ml (range, 100-600). TCWI time was 7 min (range, 4-28). FR was 4%. In three failed cases, the scope reached the ascending colon. CR was 2%. There were two perforations in the ascending colon. All three operators had similar TCWI time (p = 0.673), FR (p > 0.1), and CR (p > 0.1). A total of 98.3% of rats survived to planned sacrifice. At 48-h necropsy, the injection site was macroscopically identified in 118 rats. CONCLUSIONS: A safe and reproducible TCWI rat model has been achieved, which may provide a valuable tool in the future for studies of solid colorectal tumors. PMID- 16341567 TI - Analysis of the SAGES Outcomes Initiative groin hernia database. AB - BACKGROUND: In 1999, the Society of American Gastrointestinal Endoscopic Surgeons (SAGES) introduced the SAGES Outcomes Initiative as a way for its members to track their own outcomes. It contains perioperative and postoperative data on nearly 20,000 operations. This report provides a descriptive analysis of the groin hernia database. METHODS: The SAGES Outcomes Initiative database was accessed for all groin hernia cases from September 1999 to February 2005. The data from the preoperative, intraoperative, and postoperative entries were summarized. These data are purely descriptive and no statistical analysis was done. RESULTS: The hernia registry contains 1,607 entries, with 1,070 follow-up entries. Males comprised 85% of patients, 63% were employed, 62% had at least one comorbidity, with 84% ASA class I or II. Primary, unilateral hernia accounted for 86% of cases, whereas 14% were recurrent, 11% bilateral, 6% incarcerated, and 3% required emergency repair. The operating surgeon was the attending surgeon in 83% of cases. Anesthetic techniques were general anesthesia in 74% of cases, regional in 7%, and local in 34%, with only 16% of cases local only. Most patients had symptomatic hernias and symptoms were improved in more than 95% of patients. Most repairs were open, although 45% were endoscopic. The most frequently cited postoperative event was significant bruising (6%), with more than 99% of complications being class I or II. More than 95% of patients were able to return to work by the first postoperative visit. Patients who underwent endoscopic repair were reported to have fewer days of narcotic use than patients undergoing open repairs (0 vs 3). CONCLUSIONS: First analysis of the SAGES Outcomes Initiative groin hernia database demonstrates that (a) this is one of the largest prospective; voluntary hernia registries; (b) missing data are infrequent; and (c) the data are similar to published data from national, mandatory registries and randomized trials. Although the SAGES Outcomes Initiative is a voluntary registry, initially designed for surgeon self-assessment, and it therefore has the potential for methodological concerns inherent to voluntary registries, the findings from this first analysis are encouraging. Efforts are ongoing to simplify data entry (PDA), refine data parameters, increase surgeon participation, and determine the role of data audit and thereby the potential for clinical research. PMID- 16341568 TI - Outcome of laparoscopic Nissen fundoplication in patients with body mass index >or=35. AB - BACKGROUND: To date, few studies have examined the effect of morbid obesity on the outcome of laparoscopic antireflux surgery and results have been conflicting. The aim of this work was to study the outcome of laparoscopic Nissen fundoplication (LNF) in patients with body mass index (BMI) >or= 35. METHODS: We prospectively followed 70 patients (15 men, 55 women) with a proven diagnosis of gastroesophageal reflux disease (GERD) and a mean BMI of 38.4 +/- 0.5 (range, 35 51) who underwent LNF. All patients underwent 24-h pH study, esophageal manometry, upper gastrointestinal (GI) endoscopy, and GERD symptom score before and 6 months after LNF. Surgical outcomes were compared to those of 70 sequential nonobese patients (BMI < 30) who also underwent LNF. RESULTS: LNF was completed laparoscopically in 69 of 70 patients in the morbidly obese (MO) group and in all 70 patients in the normal-weight (NW) group. The mean operative time for the MO group was not significantly longer than that for the NW group (55.9 +/- 2.3 min vs 50.0 +/- 2.1 min), but the mean length of stay was significantly longer (3.17 +/- 0.2 days vs 2.2 +/- 0.1 days, p < 0.0001) in the MO group. There was one postoperative complication (a transhiatal herniation of the stomach) in the morbidly obese group. In both patient groups, LNF resulted in a significant increase in lower esophageal sphincter (LES) pressures. This was associated with a significant decrease in percent acid reflux in 24-h testing and a significant improvement in GERD symptom score in both groups, although patients in the MO group had a significantly higher mean reflux symptom score after surgery than did those in the NW group. After a mean follow-up of 41.6 +/- 2.9 months, one patient in the MO group required reoperation and one proton pump inhibitor therapy (PRN PPI), as required. CONCLUSIONS: Morbid obesity does not adversely affect the outcome of LNF. The conversion rate is low when performed by an experienced surgeon. PMID- 16341569 TI - Early U.S. outcomes of laparoscopic gastric bypass versus laparoscopic adjustable silicone gastric banding for morbid obesity. AB - Laparoscopic gastric bypass (LGBP) is the gold standard operation for long-term weight control in the United States. Laparoscopic adjustable silicone gastric banding (LASGB) is the preferred operative method for morbid obesity worldwide. Limited data are available comparing the two procedure in the United States. This study compares weight loss, complications, and early outcome of comorbidity resolution in patients who underwent LGBP versus LASGB. A review of prospectively collected data was performed on 392 patients undergoing primary LGBP (n = 232) and LASGB (n = 160) procedures between February 2001 and July 2004. Differences in percentage excess weight lost (%EWL) at 3, 6, 12, 18, and 24 months postop, improvement or resolution of comorbidities, and complications across procedure types were evaluated. Mean initial body mass index between groups was not significantly different (LGBP 47.2 vs LASGB 47.1, p < 0.53). There were significant differences in age, gender, and self-reported sweet-eating behavior between operative groups. There was a significantly greater %EWL in patients who underwent LGBP compared to patients of the LASGB groups 3, 6, 12, and 18 months after surgery. There were no significant differences in resolution or improvement of comorbidities between the groups. Although LGBP patients experienced more complications compared to LASGB patients (5.6 vs 4.3%, respectively; p < 0.56), this did not reach statistical significance. Early after surgery, LGBP patients lose more weight than LASGB patients but have similar improvements in comorbidities. Further follow-up is needed to determine the relative long-term efficacy of these procedures. PMID- 16341570 TI - Laparoscopic cholecystectomy using a newly developed laparoscope manipulator for 10 patients with cholelithiasis. AB - BACKGROUND: Laparoscopic surgery has continued to gain popularity in almost all fields of abdominal surgery, and robotic systems have been introduced in general surgery. Naviot is a new remote-controlled laparoscope manipulator system controlled by the operator's hand. This study assessed its introduction into clinical practice. METHODS: A group of 10 consecutive patients with cholelithiasis underwent laparoscopic cholecystectomy assisted by the Naviot system (Naviot group). Another group of 41 patients who underwent laparoscopic cholecystectomy with a conventional human camera holder (human camera group) were selected for a comparison of their operative results with those of the Naviot group. RESULTS: The operative time of 89.3 +/- 27.1 min for the Naviot group was significantly longer than that of 74.8 +/- 28.1 min for the human camera group (p < 0.05). However, when the setup time for the Naviot system was excluded, the operative time was not significantly different from that for the human camera group. Other operative results showed no significant difference between the two groups. CONCLUSIONS: The authors believe that the new Naviot system is feasible for clinical use, and that it enables surgeons to perform solo gastrointestinal surgery. PMID- 16341571 TI - Fully automated system for three-dimensional bronchial morphology analysis using volumetric multidetector computed tomography of the chest. AB - Recent advancements in computed tomography (CT) have enabled quantitative assessment of severity and progression of large airway damage in chronic pulmonary disease. The advent of fast multidetector computed tomography scanning has allowed the acquisition of rapid, low-dose 3D volumetric pulmonary scans that depict the bronchial tree in great detail. Volumetric CT allows quantitative indices of bronchial airway morphology to be calculated, including airway diameters, wall thicknesses, wall area, airway segment lengths, airway taper indices, and airway branching patterns. However, the complexity and size of the bronchial tree render manual measurement methods impractical and inaccurate. We have developed an integrated software package utilizing a new measurement algorithm termed mirror-image Gaussian fit that enables the user to perform automated bronchial segmentation, measurement, and database archiving of the bronchial morphology in high resolution and volumetric CT scans and also allows 3D localization, visualization, and registration. PMID- 16341572 TI - Oleic acid as optimizer of the skin delivery of 5-aminolevulinic acid in photodynamic therapy. AB - PURPOSE: In photodynamic therapy (PDT), topically applied aminolevulinic acid (5 ALA) is converted to protoporphyrin IX (PpIX), which upon light excitation induces tumor destruction. To optimize 5-ALA-PDT via improving the highly hydrophilic 5-ALA limited penetration into the skin, we propose the use of the known skin penetration enhancer, oleic acid (OA). METHODS: In vitro skin penetration and retention of 5-ALA (1% w/w) were measured in the presence or absence of OA (2.5, 5.0, and 10.0% w/w) in propylene glycol (PG) using porcine ear skin as the membrane. In vivo accumulation of PpIX, 4 h after application, was determined fluorometrically in healthy mice skin by chemical extraction of skin samples. In vivo PpIX fluorescence kinetics was also investigated by noninvasive techniques using an optical fiber probe, for 30 min up to 24 h after topical application of 1.0% 5-ALA + 10.0% OA in PG on hairless mice skins. RESULTS: The flux and in vitro retention of 5-ALA in viable epidermis increased in the presence of 10.0% (w/w) OA. The amounts of PpIX, evaluated both by chemical tissue extractions and in vivo measurements by an optical fiber probe, increased after applying 5-ALA formulations containing 5.0 or 10.0% OA. Moreover, in vivo kinetic studies showed an increase in skin PpIX accumulation when formulations containing 10% OA were used; PpIX accumulation was also maintained longer compared to controls. CONCLUSIONS: Both in vitro and in vivo results show the OA potential as an optimizer of 5-ALA skin delivery. PMID- 16341573 TI - Dense gas processing of micron-sized drug formulations incorporating hydroxypropylated and methylated beta-cyclodextrin. AB - PURPOSE: Because of their importance in pharmaceutical applications, hydroxypropyl-beta-cyclodextrin and methyl-beta-cyclodextrin have been selected to study the formation of micronized complexes incorporating active pharmaceutical ingredients (APIs) and cyclodextrins (CDs) by dense gas (DG) processing. METHODS: A single-step DG technique was used as an alternative to conventional methods for the manufacturing of API/CD complexes. The DG technology is highly attractive in the pharmaceutical industry because of its potential to generate micronized particles with controlled particle size distributions at moderate operating conditions. The effect of the aerosol solvent extraction system (ASES) processing on the dissolution performance of naproxen (NPX) was examined. RESULTS: The CDs were produced as microspheres smaller than 3 microm. The coprecipitation of each CD with NPX resulted in the production of microparticles with enhanced dissolution rates. CONCLUSIONS: The ASES was operated under mild conditions and generated micron-sized spherical particles that could be of particular interest in formulations for pulmonary delivery. Particular advantages of the technique are that (1) nontoxic solvents are used, and (2) it is suitable for the processing of thermally labile compounds. The proposed process can create opportunities to improve current administration routes and exploit novel delivery systems for drug formulations incorporating CDs. PMID- 16341574 TI - Smad3 specific inhibitor, naringenin, decreases the expression of extracellular matrix induced by TGF-beta1 in cultured rat hepatic stellate cells. AB - PURPOSE: During the process of liver fibrogenesis, transforming growth factor beta (TGF-beta) plays an essential role in modulating extracellular matrix (ECM) gene expression, and a growing body of evidence suggests that this is a Smad3 dependent process in the activated hepatic stellate cells (HSCs). Naringenin showed a significantly protective effect on experimental rat liver fibrosis, in our efforts to elucidate its antifibrosis molecular mechanisms and to find a novel target based on Smad3 signaling for challenging fibrosis diseases. METHODS: In this study, reverse transcription-polymerase chain reaction and Western blot assays were used to investigate the inhibitory effect of naringenin on ECM formation induced by TGF-beta1 in the HSC-T6 cells. RESULTS: Naringenin reduced not only the accumulation of ECM, including collagen Ialpha1 (Col Ialpha1), fibronectin (FN), and plasminogen activator inhibitor-1 (PAI-1), but also the production of Smad3 induced by TGF-beta1 in both mRNA and protein levels in a dose-dependent manner. Moreover, naringenin selectively inhibited the transcription of Smad3, but not other Smads involved in TGF-beta1 signaling pathways. CONCLUSION: Our data demonstrate that naringenin can exert antifibrogenic effects by directly or indirectly down-regulating Smad3 protein expression and phosphorylation through TGF-beta signaling. PMID- 16341575 TI - Modularity and intersection of "what", "where" and "how" processing of visual stimuli: a new method of FMRI localization. AB - Research on the modularity of perceptual and cognitive processes has often pointed to a ventral-dorsal distinction in cortical pathways that depend upon the nature of the stimuli and the task. However, it is not clear whether the dorsal, occipital-parietal stream specializes in locating visual objects (i.e., a "where" stream), or taking action toward objects (i.e., a "how" stream), although there is some consensus for a ventral, occipital-temporal "what" stream that specializes in the identification of visual objects. It is also not clear to what extent word and picture processing are modular along these streams, as functional imaging maps to date have not addressed the modularity question directly. Here we present two types of functional imaging maps that directly show modularity and intersection of processing function for word and picture stimuli in tasks that require decisions about "what is", "where is", or "how do you interact with" a stimulus (N=6 participants). Our results reveal a middle dorsal "how" stream with some modular regions of activation that are distinct from activation during "where" processing, and that words and pictures involve several modular regions of activation along these streams. PMID- 16341576 TI - Source localization of early stages of face processing. AB - Recent studies using ERPs in face recognition revealed that face processing starts around 100 ms after stimulus onset, 70 ms earlier than suggested before. While the neural sources of the N 170 component have repeatedly been found to be localized in the gyrus fusiformis and the inferior occipital cortex, sources have not yet been investigated for the P100 component during face processing. Therefore, we measured the ERPs elicited by faces and control stimuli in 72 subjects in order to localize the neural sources of both the P100 and the N 170 component. We observed significantly higher P100 and N 170 amplitudes to faces compared to control stimuli. LORETA source localization revealed significantly higher brain activity in the left and right gyrus fusiformis for the N 170 component, with additional regions of increased brain activation in a parieto temporal-occipital network. For the P100, faces activated the left and right gyrus fusiformis significantly stronger than control stimuli. This study reveals that the first step of face processing (about 100 ms after stimulus presentation) is localized in the gyrus fusiformis. The second step of face processing around 170 ms involves the gyrus fusiformis, with additional activation in a more distributed network, including the occipital cortex. PMID- 16341577 TI - Linear inverse solutions: simulations from a realistic head model in MEG. AB - Distributed linear solutions are widely used in source localization to solve the ill-posed EEG/MEG inverse problem. In the classical approach based on dipole sources, these methods estimate the current densities at a great number of brain sites, typically at the nodes of a 3-D grid which discretizes the chosen solution space. The estimated current density distributions are displayed as brain electromagnetic tomography (BET) images. We have tested well known minimum norm solutions (MN, WMN, LORETA) and other linear inverse solutions [WROP, sLORETA, interference uniform, gain uniform, weight vector normalized (WVN), and a new solution named SLF (Standardized Lead Field)], using a MEG configuration (BTi Magnes 2500 WH with 148 axial magnetometers) and a realistic head model using BEM (Boundary Element Method). The solutions were compared in a noise-free condition and in the presence of noise using the classical dipole localization errors (DLE) together with a new figure of merit that we called max gain uniformity, which measures the capability of an inverse linear solution to show spots of activity with similar amplitudes on the brain electromagnetic tomographies when multiple dipole sources with similar moments are simultaneously active. Whereas some solutions (sLORETA, interference uniform and SLF) were capable of zero dipole localization errors in the noise-free case, none of them reached 100% of correct dipole localizations in the presence of a high level of Gaussian noise. The SLF solution, which has the advantage to be independent from any regularization parameter, presented the best results with the lowest max gain uniformities, with almost 100% of correct dipole localizations with 10% of noise and more than 90% of correct localizations with 30% of noise added to the data. Nevertheless, no solution was able to combine at the same time a correct localization of single sources and the capability to visualize multiple sources with comparable amplitudes on the brain electromagnetic tomographies. PMID- 16341578 TI - Integrated MEG and fMRI model: synthesis and analysis. AB - An integrated model for magnetoencephalography (MEG) and functional Magnetic Resonance Imaging (fMRI) is proposed. In the model, the neural activity is related to the Post Synaptic Potentials (PSPs) which is common link between MEG and fMRI. Each PSP is modeled by the direction and strength of its current flow which are treated as random variables. The overall neural activity in each voxel is used for equivalent current dipole in MEG and as input of extended Balloon model in fMRI. The proposed model shows the possibility of detecting activation by fMRI in a voxel while the voxel is silent for MEG and vice versa. Parameters of the model can illustrate situations like closed field due to non-pyramidal cells, canceling effect of inhibitory PSP on excitatory PSP, and effect of synchronicity. In addition, the model shows that the crosstalk from neural activities of the adjacent voxels in fMRI may result in the detection of activations in these voxels that contain no neural activities. The proposed model is instrumental in evaluating and comparing different analysis methods of MEG and fMRI. It is also useful in characterizing the upcoming combined methods for simultaneous analysis of MEG and fMRI. PMID- 16341579 TI - Distribution of spatial complexity of EEG in idiopathic generalized epilepsy and its change after chronic valproate therapy. AB - The objective of this study was to investigate the global and regional spatial synchrony of the EEG background activity, and to assess the effect of chronic valproate therapy on spatial synchrony. 15 idiopathic generalized epilepsy (IGE) patients were examined and compared to 16 normal controls. Resting EEG with 19 channels was investigated before and during chronic administration of valproate (VPA). Omega, a single-valued measure of spatial covariance complexity, was calculated to assess the degree of spatial synchrony of EEG. Furthermore, a new parameter was defined to characterize the distribution of spatial synchrony (Antero-Posterior Complexity Ratio, APCR). Global Omega complexity was significantly lower in IGE compared to controls, while regional complexity showed significant differences only in the anterior region: the IGE group showed lower complexity. APCR was significantly lower in IGE. VPA therapy (1) lowered the global complexity, (2) increased regional complexity in the anterior region, but decreased it in the posterior region, and (3) increased APCR. In IGE lower complexity, i.e. enhanced spatial synchrony, was found, especially in the anterior cortical area. VPA modified the distribution of spatial synchrony in IGE patients towards that of normal controls, although the effect is not identical with full normalization of cortical bioelectric activity. Whether the observed change of spatial synchrony distribution may reflect the normalizing effect of valproate on the brain state is worth further investigation. PMID- 16341583 TI - The phenomenon of "pre-ischaemic conditioning" in the brain only partly involves the NMDA receptor: a magnetic resonance study. AB - We have investigated in more detail our previous observations on a form of ischaemic pre-conditioning "metabolic adaptation", i.e.--that sequential metabolic insults (hypoxia followed 40 min later by combined hypoxia + hypoglycaemia, or vice versa) are less injurious (monitored by increased [Ca2+]i and decreased PCr) than the immediate combined insult. We have now observed that the "adaptation" occurs between 10 and 20 min. Pre-treatment of the tissues with 10 microM-MK801 showed that it had no effect on the increase in [Ca2+]i caused by the sequential insult and only partially blocked the increase observed by exposure to the immediate combined insult. Exposure to both the delayed and immediate combined insults with low extracellular Ca2+ resulted in a two-fold increase in [Ca2+]i, similar to the increase observed with normal extracellular Ca2+ in the presence of MK801. The results are discussed in terms of the possible origins of the increases in [Ca2+]i. PMID- 16341582 TI - The role of glial adenosine receptors in neural resilience and the neurobiology of mood disorders. AB - Adenosine receptors were classified into A1- and A2-receptors in the laboratory of Bernd Hamprecht more than 25 years ago. Adenosine receptors are instrumental to the neurotrophic effects of glia cells. Both microglia and astrocytes release after stimulation via adenosine receptors factors that are important for neuronal survival and growth. Neuronal resilience is now considered as of pivotal importance in the neurobiology of mood disorders and their treatment. Both sleep deprivation and electroconvulsive therapy, two effective therapeutic measures in mood disorders, are associated with an increase of adenosine and upregulation of adenosine A1-receptors in the brain. Parameters closely related to adenosine receptor activation such as cerebral metabolic rate and delta power in the sleep EEG provide indirect evidence that adenosinergic signaling may be associated with the therapeutic response to these measures. Thus, neurotrophic effects evoked by adenosine receptors might be important in the mechanism of action of ECT and perhaps also sleep deprivation. PMID- 16341584 TI - Ependymal cell differentiation and GLUT1 expression is a synchronous process in the ventricular wall. AB - Ependymal cells appear to be totally differentiated during the first 3 weeks in the mouse brain. Early during postnatal development ependymal cells differentiate and undergo metabolic activation, which is accompanied by increased glucose uptake. We propose that ependymal cells induce an overexpression of the glucose transporter, GLUT1, during the first 2 weeks after delivery in order to maintain the early metabolic activation. During the first postnatal day, GLUT1 is strongly induced in the upper region of the third ventricle and in the ventral area of the rostral cerebral aqueduct. During the next 4 days, GLUT1 is expressed in all differentiated ependymal cells of the third ventricle and in hypothalamic tanycytes. At the end of the first week, ependymal cell differentiation and GLUT1 overexpression is concentrated in the latero-ventral area of the aqueduct. We propose that ependymal cell differentiation and GLUT1 overexpression is a synchronous process in the ventricular wall. PMID- 16341585 TI - Expression of multidrug resistance proteins (Mrps) in astrocytes of the mouse brain: a single cell RT-PCR study. AB - Multidrug resistance proteins (Mrps) are ATP-driven export pumps which mediate the export of organic anions such as glutathione conjugates and glucuronides from eukaryotic cells. Within the central nervous system astrocytes have important functions in metabolism and detoxification. In such processes Mrps play essential roles. To identify the Mrp repertoire of mouse brain and of astrocytes in particular, the expression of six mouse Mrps was investigated by reverse transcription polymerase-chain reaction (RT-PCR). Using mouse brain mRNA as source, amplification products were obtained for Mrp1, Mrp3, Mrp4, Mrp5 and Mrp6. In contrast, mRNA of Mrp2 could not be detected in mouse brain. To investigate whether individual astrocytes express different Mrps in brain, single-cell RT PCRs were performed from the cytosol harvested from single astrocytes in acutely isolated brain slices from cortex and cerebellum of TgN(GFAP-EGFP) mice. In these mice astrocytes can readily be identified by glial fibrillary acidic protein promoter-controlled green fluorescent protein expression. Investigation of individual cortical astrocytes and Bergmann glial cells revealed that these cells express Mrp1, Mrp4 and Mrp5 and that individual astrocytes can contain mRNA of one, two or three of these Mrps simultaneously. PMID- 16341586 TI - Mitochondrial aconitase is a transglutaminase 2 substrate: transglutamination is a probable mechanism contributing to high-molecular-weight aggregates of aconitase and loss of aconitase activity in Huntington disease brain. AB - Transglutaminase activity was found to be present in highly purified non synaptosomal rat brain mitochondria. A 78-kDa protein in these organelles was shown to be a transglutaminase 2 substrate, and incubation of a non-synaptosomal mitochondrial lysate with transglutaminase 2 yielded high-Mr proteins. The 78-kDa protein was identified as mitochondrial aconitase by MALDI-TOF analysis. Aconitase activity was decreased in a dose-dependent manner when non-synaptosomal rat brain mitochondria were incubated with transglutaminase 2. Transglutaminase activity is increased about 2-fold in the mitochondrial fraction of HD caudate. Moreover, Western blotting of the mitochondrial fraction revealed that most of the mitochondrial aconitase in HD caudate is present as high-Mr aggregates. Aconitase activity was previously shown to be decreased in Huntington disease (HD) caudate (a region severely damaged by the disease). The present findings suggest that an increase of transglutaminase activity in HD caudate may contribute to mitochondrial dysfunction by incorporating aconitase into inactive polymers. PMID- 16341587 TI - Translocation of ethanolamine phosphoglyceride is required for initiation of apoptotic death in OLN-93 oligodendroglial cells. AB - The possible interplay between extracellular signal-regulated protein kinase (ERK) activation and ethanolamine phosphoglycerides (PG) membrane bilayer translocation following oxidative stress (OS) (0.5 mM H2O2/0.05 mM Fe2+), was examined in oligodendroglia, OLN93, cells with altered plasma membrane PG composition. Cells supplemented with 50 microM docosahexaenoic acid (DHA, 22:6n3) to increase the number of potential double bond targets for OS in ethanolamine-PG (EPG) were compared to cells with diminished content of EPG, attained by the addition of 0.5 mM N,N-dimethylethanolamine (dEa). After 30 min OS, EPG translocation accompanied by sustained ERK activation and nuclear translocation culminating in apoptosis was found in DHA-supplemented cells in contrast to no EPG translocation, a brief ERK activation, but no nuclear translocation, and no cell death in DHA/dEa-supplemented cells. DHA/dEa-supplemented cells pretreated with the protein-tyrosine phosphatases inhibitor Na3VO4 followed by OS, although expressing a sustained ERK activation and nuclear translocation, failed to show apoptosis and lacked EPG translocation. In DHA-supplemented cells U0126, a MEK inhibitor, prevented ERK activation and EPG translocation and protected from cell death. These findings most likely indicate that ERK activation is an indispensable component for the signaling cascades leading to EPG translocation but only activation of the latter is leading to OS-induced apoptotic cell death. PMID- 16341588 TI - Contribution of extracellular glutamine as an anaplerotic substrate to neuronal metabolism: a re-evaluation by multinuclear NMR spectroscopy in primary cultured neurons. AB - Multinuclear NMR spectroscopy is used to investigate the effect of glutamine on neuronal glucose metabolism. Primary neurons were incubated with [1-(13C)]glucose in the absence or presence of glutamine (2 mM) and/or NH4Cl (5 mM). After ammonia treatment, the concentrations of high-energy phosphates decreased up to 84% of control, which was aggravated in glutamine-containing medium (up to 42% of control). These effects could not be attributed to changes in mitochondrial glucose oxidation. Withdrawal of glutamine decreased amino acid concentrations, e.g. of glutamate to 53%, but also considerably lessened the 13C enrichment in [4 (13C)]glutamate to 8.3% of control, and decreased the 13C-enrichment in acetyl CoA entering the Krebs cycle (P < 0.001). Thus, although glutamine is potent in replenishing neuronal glutamate stores, glutamate formation is mainly attributed to its de novo synthesis from glucose. Furthermore, mitochondrial glucose metabolism strongly depends on the supply of carbons from glutamine, indicating that exogenous glutamine is a well-suited substrate to replenish neuronal Krebs cycle intermediates. PMID- 16341589 TI - Carnosine protects the brain of rats and Mongolian gerbils against ischemic injury: after-stroke-effect. AB - Carnosine, a specific constituent of excitable tissues of vertebrates, exhibits a significant antioxidant protecting effect on the brain damaged by ischemic reperfusion injury when it was administered to the animals before ischemic episode. In this study, the therapeutic effect of carnosine was estimated on animals when this drug was administered intraperitoneally (100 mg/kg body weight) after ischemic episode induced by experimental global brain ischemia. Treatment of the animals with carnosine after ischemic episode under long-term (7-14 days) reperfusion demonstrated its pronounced protective effect on neurological symptoms and animal mortality. Carnosine also prevented higher lipid peroxidation of brain membrane structures and increased a resistance of neuronal membranes to the in vitro induced oxidation. Measurements of malonyl dialdehyde (MDA) in brain homogenates showed its increase in the after brain stroke animals and decreased MDA level in the after brain stroke animals treated with carnosine. We concluded that carnosine compensates deficit in antioxidant defense system of brain damaged by ischemic injury. The data presented demonstrate that carnosine is effective in protecting the brain in the post-ischemic period. PMID- 16341590 TI - Identification of a mouse synaptic glycoprotein gene in cultured neurons. AB - Neuronal differentiation and aging are known to involve many genes, which may also be differentially expressed during these developmental processes. From primary cultured cerebral cortical neurons, we have previously identified various differentially expressed gene transcripts from cultured cortical neurons using the technique of arbitrarily primed PCR (RAP-PCR). Among these transcripts, clone 0-2 was found to have high homology to rat and human synaptic glycoprotein. By in silico analysis using an EST database and the FACTURA software, the full-length sequence of 0-2 was assembled and the clone was named as mouse synaptic glycoprotein homolog 2 (mSC2). DNA sequencing revealed transcript size of mSC2 being smaller than the human and rat homologs. RT-PCR indicated that mSC2 was expressed differentially at various culture days. The mSC2 gene was located in various tissues with higher expression in brain, lung, and liver. Functions of mSC2 in neurons and other tissues remain elusive and will require more investigation. PMID- 16341591 TI - Compartmentation of lactate originating from glycogen and glucose in cultured astrocytes. AB - Brain glycogen metabolism was investigated by employing isofagomine, an inhibitor of glycogen phosphorylase. Cultured cerebellar and neocortical astrocytes were incubated in medium containing [U-(13C)]glucose in the absence or presence of isofagomine and the amounts and percent labeling of intra- and extracellular metabolites were determined by mass spectrometry (MS). The percent labeling in glycogen was markedly decreased in the presence of isofagomine. Surprisingly, the percent labeling of intracellular lactate was also decreased demonstrating the importance of glycogen turnover. The decrease was limited to the percent labeling in the intracellular pool of lactate, which was considerably lower compared to that observed in the medium in which it was close to 100%. These findings indicate compartmentation of lactate derived from glycogenolysis and that derived from glycolysis. Inhibiting glycogen degradation had no effect on the percent labeling in citrate. However, the percent labeling of extracellular glutamine was slightly decreased in neocortical astrocytes exposed to isofagomine, indicating an importance of glycogen turnover in the synthesis of releasable glutamine. In conclusion, the results demonstrate that glycogen in cultured astrocytes is continuously synthesized and degraded. Moreover, it was found that lactate originating from glycogen is compartmentalized from that derived from glucose, which lends further support to a compartmentalized metabolism in astrocytes. PMID- 16341592 TI - Hippocampal cannabinoid-1 receptor upregulation upon endothelin-B receptor deficiency: a neuroprotective substitution effect? AB - Endothelin (ETB)-receptors mediate anti-apoptotic actions. Lack of functional ETB receptors leads to increased neuronal apoptosis in the hippocampus. The increased apoptosis must be compensated by other mechanisms, however, as ETB-deficient rats display normal overall brain morphology. To illuminate on brain plasticity in ETB receptor deficiency, we studied the expression and function of another neuroprotective system, the cannabinoid CB1-receptors, in ETB-deficient hippocampus. We show that CB1 expression in hippocampus increases postnatally in all rats but that the increase in CB1-receptor expression is significantly higher in ETB-deficient compared to wildtype littermates. Neuronal apoptosis decreases during brain maturation but remains on a significantly higher level in the ETB deficient compared to wildtype dentate. When investigating survival of hippocampal neurons in culture, we found significant protection against hypoxia induced cell death with CB1-analogs (noladin, (9-tetrahydrocannabinol) only in ETB-deficient neurons. We suggest that CB1-receptor upregulation in the ETB mutant hippocampus reflects an attempt to compensate for the lack of ETB receptors. PMID- 16341593 TI - Astrocytes protect neurons from ammonia toxicity. AB - Ammonia is a neurotoxin that is implicated in the CNS dysfunction associated with hepatic encephalopathy, urea cycle disorders, Reye's syndrome and other neurological conditions. While in vivo studies suggest that astrocytes are the principal target of ammonia toxicity, recent in vitro investigations suggest that neurons may also be directly affected by ammonia. To further examine the issue of neural cell sensitivity to ammonia, pure rat cortical neuronal cultures, as well as co-cultures of neurons and astrocytes, were exposed to 5 mM NH4Cl for 48 h. Cultures were examined for morphological changes by light microscopy, measures of cell death, free radical production and changes in the mitochondrial inner membrane potential. Ammonia caused extensive degenerative changes in pure cultured neurons, while such neuronal changes were minor in the co-cultures. Similarly, processes of pure cultured neurons displayed a significant loss of the mitochondrial inner membrane potential, as compared to neurons in co-cultures. Cell death (LDH release) in ammonia-treated neuronal cultures was twice as great as untreated controls, while in co-cultures ammonia did not significantly increase cell death. Free radical production at 3 min was increased (69%, P<0.05) in pure neuronal cultures but not in co-cultures. The neuroprotective effects observed in co-cultures may have been mediated by the astrocyte's ability to scavenge free radicals, by their detoxification of ammonia and/or by their neurotrophic actions. The neuroprotective action of astrocytes may explain the failure to detect significant pathological changes in neurons in ammonia toxicity in vivo. PMID- 16341594 TI - p42(IP4)/centaurin alpha1, a brain-specific PtdIns(3,4,5)P3/Ins(1,3,4,5)P4 binding protein: membrane trafficking induced by epidermal growth factor is inhibited by stimulation of phospholipase C-coupled thrombin receptor. AB - The brain-specific 42-kDa protein, p42(IP4), contains a N-terminal zinc finger (ZF) motif and a tandem of two pleckstrin homology (PH) domains. p42(IP4) binds in vitro the second messengers phosphatidylinositol(3,4,5)trisphosphate (PtdIns(3,4,5)P3) and inositol(1,3,4,5)tetrakisphosphate (Ins(1,3,4,5)P4). We observed by confocal microscopy in live HEK 293 cells the GFP-p42(IP4), a chimera of human p42(IP4) and green fluorescence protein (GFP). There, we studied the influence of thrombin, which raises Ins(1,3,4,5)P4, on membrane translocation of GFP-p42(IP4), induced by epidermal growth factor (EGF). Thrombin in the presence of LiCl inhibited the EGF-induced membrane recruitment of GFP-p42(IP4). In the absence of LiCl, thrombin weakened the EGF-mediated membrane recruitment of GFP p42(IP4). Furthermore, the participation of p42(IP4) protein domains on the EGF mediated membrane translocation was analyzed. We used several p42(IP4) variants, in which one of the domains was deleted. Alternatively, single p42(IP4) domain GFP fusion proteins were generated. Only the p42(IP4) variant lacking the ZF domain showed a very weak membrane translocation in response to EGF stimulation, but all the other p42(IP4) variants did not translocate. Thus, we conclude that the combination of both PH domains with ZF is required for membrane translocation of p42(IP4). PMID- 16341595 TI - Dual effect of pyruvate in isolated nerve terminals: generation of reactive oxygen species and protection of aconitase. AB - Generation of reactive oxygen species (ROS) in synaptosomes was investigated in the presence of different substrates. When pyruvate was used as a substrate an increased rate of hydrogen peroxide formation was detected by the Amplex Red fluorescent assay, but aconitase, which is known to be a highly sensitive enzyme to ROS was not inhibited. In contrast, pyruvate exerted a partial protection on aconitase against a time-dependent inactivation that occurred when synaptosomes were incubated in the absence of substrates. Disruption of synaptosomal membranes with Triton X-100 prevented the protective effect of pyruvate. It is suggested that citrate and/or isocitrate formed in the metabolism of pyruvate could be responsible for a partial protection of aconitase. Therefore while pyruvate could have a prooxidant effect it could also exert a protective effect on the aconitase. PMID- 16341596 TI - Carnosine and homocarnosine, the forgotten, enigmatic peptides of the brain. AB - Carnosine (beta-alanyl-histidine) and homocarnosine (gamma-aminobutyryl histidine) are major constituents of excitable tissues, brain and skeletal muscles, but their physiological functions are yet unknown. Using primary cell culture systems, synthesis and uptake of carnosine exclusively by glial cells could be demonstrated. Uptake of carnosine was found to be mediated by a high affinity, energy-dependent dipeptide transport system, subsequently identified as the peptide transporter PepT2. With the synthesis of beta-Ala-Lys-Nepsilon-AMCA as a fluorescent reporter molecule, accumulation of this dipeptide derivative could be monitored under viable conditions not only in astroglia cells but also in folliculostellate cells of the anterior pituitary and in gonadal resident macrophages. This reporter dipeptide provided a most valuable tool to identify an intrapituitary communication system by tracing folliculostellate cells in acute slice preparation. Moreover, this substance could also be used to prepare pituitary cell cultures enriched with or depleted of folliculostellate cells that are needed for further studies. PMID- 16341597 TI - You can't go home again: transcriptionally driven alteration of cell signaling by NGF. AB - Here we review findings indicating that neurotrophins such as NGF promote changes in gene transcription that in turn influence the ways that cells subsequently respond to trophic factors. As a result, initial responses of "naive" cells to NGF and other trophic agents differ from those of cells with prior NGF exposure. We discuss specific examples based on reports in the literature as well as on data derived from a serial analysis of gene expression (SAGE) study of NGF promoted transcriptional changes in PC12 pheochromocytoma cells. PMID- 16341599 TI - The utility of cumulative meta-analysis: application to programs for reducing sexual violence. AB - Recent advances in meta-analytic techniques provide a useful framework for interpreting the findings of individual studies. Simple formula are presented for determining whether the results of a single study are statistically different from the cumulative average of previous studies, and for calculating the new cumulative average. When applied to two controversial social policies (treatment of sexual offenders; rape prevention programs for college women), cumulative meta analyses suggested patterns that were not identified by the authors of the individual studies nor by narrative reviews of these content areas. PMID- 16341600 TI - Predictors of treatment attrition as indicators for program improvement not offender shortcomings: a study of sex offender treatment attrition. AB - This study classified potential attrition predictors under the domains of risk, need and responsivity (D. Andrews & J. Bonta, 2003). Non-sexual criminogenic needs (e.g. aggression, rule violating behaviors) and responsivity factors (e.g. lack of motivation and denial) were the two main clusters of predictors that correctly classified 95.3% of program completers and non-completers using discriminant function analysis in a sample of high-risk male sexual offenders treated in an accredited inpatient sex offender treatment program. Rapists were more aggressive than other types of sex offenders and were more likely to drop out of treatment. Some studies of predictors of treatment attrition have used offender problem behaviors or psychopathologies to predict attrition and then use the information to exclude offenders from treatment. Others have argued, and we concur, that results of attrition research should not be used to develop an "attrition profile" to exclude offenders from treatment. Predictors of attrition should be seen as markers for program improvement, rather than shortcomings of the offender. Suggestions for program improvements to reduce the rate of attrition, based on results of research, are presented. PMID- 16341601 TI - Youth with problematic sexualized behaviors in the child welfare system: a one year longitudinal study. AB - This study assessed continuity of problematic sexualized behaviors (PSB) over a 1 year period. Ninety-seven 10-12-year-olds in either foster boarding homes or a residential treatment center participated at Time 1. Twelve months later, 78 youth were available for a second data collection assessment. At both data collection phases, researchers interviewed foster parents or primary therapists about the youths' sexual behavior. Findings revealed significant continuity in PSB over time, with children who at Time 1 exhibited PSB significantly more likely to exhibit PSB at Time 2. The reverse was also true in that the absence of PSB at Time 1 was associated with the absence of PSB at Time 2. In addition, a subset of specific PSB behaviors was noted to be most stable, although this varied across the groups. Youth with PSB exhibited several patterns of persistence in specific behaviors over time, including continuity, a mix of continuity and change, and complete discontinuity. The persistence of PSB over time was most true for the children living in a residential treatment center, the more disturbed group studied. We conclude that the persistence of PSB is more likely when the child has other problematic behaviors. PMID- 16341602 TI - Relapse prevention with intellectually disabled sexual offenders. AB - The adaptation of relapse prevention theory to sexual offending (W. D. Pithers, J. K. Marques, C. C. Gibat, & G. A. Marlatt, 1983) has represented an important movement in cognitive-behavioural treatment for sexual offenders. However, this model of relapse prevention has been criticised for its limited view and oversimplification of the relapse prevention process (R. K. Hanson, 2000; T. Ward & S. M. Hudson, 1996). As a result, T. Ward and S. M. Hudson (2000a) have developed a multiple pathway model of the relapse prevention process based on self-regulation theory. Although this model continues to be empirically validated on sexual offenders (J. A. Bickley & A. R. Beech, 2002; T. Ward, S. M. Hudson, & J. C. McCormick, 1999), there has been no empirical research regarding the application of this theory to intellectually disabled sexual offenders. This paper discusses whether the characteristics of offenders in each of the relapse offence pathways, as described by T. Ward and S. M. Hudson (2000a), may be similar to the characteristics of intellectually disabled sexual offenders. From a review of the literature, it appears that the intellectually disabled sexual offender may be most likely to offend via the approach-automatic pathway or the avoidant-passive pathway. The potential treatment implications of the self regulation model for intellectually disabled sexual offenders is discussed, as well as the need for empirical evaluation with regards to the application of this model to the intellectually disabled sexual offender population. PMID- 16341603 TI - Identifying schemas in child molesters, rapists, and violent offenders. AB - One focus of research concerning offending behavior has been the concept of the cognitive distortion, although the importance of the need for research into the underlying cognitive structures in offenders has been highlighted. This study examined schemas in child molesters, rapists, and violent offenders, predicting content differences in the offence-related schemas between these groups. In accordance with previous research, the prevalence of a "suspiciousness/hostility to women" schema in rapists was predicted. Twelve rapists, twelve child molesters, and twelve violent offenders incarcerated in a male maximum-security prison completed "Life Maps" (autobiographies) and the My Life questionnaire (R. E. Mann & C. R. Hollin, 2001), both designed to indicate the presence of schemas. A content analysis template containing nine schemas was constructed and applied to the Life Map data. Results showed a significant difference in the prevalence of schema type between the three groups. There was a prevalence of a "hostility/distrust of women" schema in the rapists. Analysis of the My Life questionnaire revealed a difference between violent offenders and rapists only. This study highlights the need for further research into schemas with sexual and violent offenders. PMID- 16341604 TI - The strength of sexual arousal as a function of the age of the sex offender: comparisons among pedophiles, hebephiles, and teleiophiles. AB - Previous research has shown that sexual arousability in human males declines from its peak in early adolescence until old age. This study compared the rates of decline in three groups of males: those most attracted sexually to prepubescent children (pedophiles), those most attracted to pubescent children (hebephiles), and those most attracted to physically mature persons (teleiophiles). The participants were 2,028 patients referred to Toronto's Centre for Addiction and Mental Health from 1995 to 2004 for evaluation of criminal or otherwise disturbing sexual behavior, but not for erectile or ejaculatory problems. All underwent phallometric assessment for erotic age-preference. This is a psychophysiological technique in which an individual's penile blood volume is monitored while he is presented with a standardized set of laboratory stimuli depicting male and female children, pubescents, and adults. The experimental measure of sexual arousability was the average of the participant's three greatest penile responses to any stimulus category, expressed in cubic cm of blood volume increase. The results showed that sexual arousability was an inverse function of age, and that there were no differences between the pedophiles, hebephiles, and teleiophiles in the rate at which arousability declined. PMID- 16341605 TI - [Cost of illness studies as a basis for decision making]. AB - Definitions and methods for cost of illness studies are explained: costs, direct costs, indirect costs, intangible costs, the human capital method and the willingness to pay approach. Devaluing the long-term health benefits from prevention by constant discount rates is questioned. The development and the state of the art of cost of illness studies at a national level in Germany are discussed. As an example the newly developed cost of illness study of the Federal Statistical Office with direct and indirect costs of circulatory diseases is presented. Reasons for the benefit of cost of illness studies for decision making are given in the context of costs of illness and aging. PMID- 16341606 TI - [Health economics of chronic infectious diseases: the example of hepatitis C]. AB - Based on the German Hepatitis C Model (GEHMO) we developed a Hepatitis C Policy Model and applied it to the heterogeneous German hepatitis C population within the German health care context. We used Markov cohort simulation to predict absolute clinical and economic outcomes for a 20-year time horizon. For the cost effectiveness analysis, a lifelong time horizon was used. Four different strategies were compared: (1) no antiviral treatment, (2) interferon monotherapy, (3) combination therapy with interferon plus ribavirin, and (4) combination therapy with pegylated interferon plus ribavirin. Based on our model, antiviral therapy with pegylated interferon and ribavirin could prevent about 17,000 cases of cirrhosis, 580 liver transplants, and 7,600 HCV-related deaths and is expected to save about 53,000 life years at total costs of 1.3 billion Euros within the next 20 years. Pegylated interferon plus ribavirin was the most effective treatment with an incremental cost-utility ratio of 23,000 Euros per quality adjusted life year saved. PMID- 16341607 TI - [Aspects of economic evaluation in health care]. AB - Health economists use models to estimate comparative costs and usage of interventions in health care. However the concepts and methods used have inherent weaknesses, especially in the determination of relevant and exact effect sizes. Health economic methodologies do not replace sociopolitical decision making, but they can play an important role in rational decision making about necessary changes to our social and health system if the instruments themselves are neither over- nor underestimated. PMID- 16341608 TI - Streptomyces daliensis sp. nov. from soil. AB - A novel actinomycete strain YIM 31724(T) was isolated from a soil sample collected from Dali, Yunnan Province, People's Republic of China. The strain is characterized by white to yellow white aerial mycelia, spiral spore chains and smooth spore surface. The cell wall of strain YIM 31724(T) contained LL diaminopimelic acid (A(2)pm) and traces of meso-A(2)pm. Whole-cell hydrolysates contained mainly glucose and small amounts of galactose and xylose. The menaquinones were MK-9(H(6)) (31%) and MK-9(H(8)) (69%). Phosphatidylethanolamine was the diagnostic phospholipid. The DNA G+C content of strain YIM 31724(T) was 67.2 mol%. Phylogenetic analysis indicated that the strain belongs to the genus Streptomyces, with highest similarity to Streptomyces rimosus subsp. rimosus JCM 4667(T) (rRNA gene sequence similarity value of 98.9%) and Streptomyces erumpens DSM 40941(T) (rRNA gene sequence similarity value of 98.7%). Based on its phenotypic and genotypic characteristics, including low DNA-DNA hybridization results, strain YIM 31724(T)(= CCTCC AA204020(T) = KCTC 19076(T)) is proposed as the type strain of a novel species, Streptomyces daliensis sp. nov. PMID- 16341609 TI - Replicated linkage for eye color on 15q using comparative ratings of sibling pairs. AB - : The aim of the study was to perform a genetic linkage analysis for eye color, for comparative data. Similarity in eye color of mono- and dizygotic twins was rated by the twins' mother, their father and/or the twins themselves. For 4,748 twin pairs the similarity in eye color was available on a three point scale ("not at all alike"-"somewhat alike"-"completely alike"), absolute eye color on individuals was not assessed. The probability that twins were alike for eye color was calculated as a weighted average of the different responses of all respondents on several different time points. The mean probability of being alike for eye color was 0.98 for MZ twins (2,167 pairs), whereas the mean probability for DZ twins was 0.46 (2,537 pairs), suggesting very high heritability for eye color. For 294 DZ twin pairs genome-wide marker data were available. The probability of being alike for eye color was regressed on the average amount of IBD sharing. We found a peak LOD-score of 2.9 at chromosome 15q, overlapping with the region recently implicated for absolute ratings of eye color in Australian twins [Zhu, G., Evans, D. M., Duffy, D. L., Montgomery, G. W., Medland, S. E., Gillespie, N. A., Ewen, K. R., Jewell, M., Liew, Y. W., Hayward, N. K., Sturm, R. A., Trent, J. M., and Martin, N. G. (2004). Twin Res. 7:197-210] and containing the OCA2 gene, which is the major candidate gene for eye color [Sturm, R. A. Teasdale, R. D, and Box, N. F. (2001). Gene 277:49-62]. Our results demonstrate that comparative measures on relatives can be used in genetic linkage analysis. PMID- 16341610 TI - Genome-wide scan of IQ finds significant linkage to a quantitative trait locus on 2q. AB - A genome-wide linkage scan of 795 microsatellite markers (761 autosomal, 34 X chromosome) was performed on Multidimensional Aptitude Battery subtests and verbal, performance and full scale scores, the WAIS-R Digit Symbol subtest, and two word-recognition tests (Schonell Graded Word Reading Test, Cambridge Contextual Reading Test) highly predictive of IQ. The sample included 361 families comprising 2-5 siblings who ranged in age from 15.7 to 22.2 years; genotype, but not phenotype, data were available for 81% of parents. A variance components analysis which controlled for age and sex effects showed significant linkage for the Cambridge reading test and performance IQ to the same region on chromosome 2, with respective LOD scores of 4.15 and 3.68. Suggestive linkage (LOD score>2.2) for various measures was further supported on chromosomes 6, 7, 11, 14, 21 and 22. Where location of linkage peaks converged for IQ subtests within the same scale, the overall scale score provided increased evidence for linkage to that region over any individual subtest. Association studies of candidate genes, particularly those involved in neural transmission and development, will be directed to genes located under the linkage peaks identified in this study. PMID- 16341611 TI - Thyroid endocrine disruption in stonerollers and cricket frogs from perchlorate contaminated streams in east-central Texas. AB - In October 2001 and March 2002, a field survey of central stonerollers (Campostoma anomalum) from perchlorate-contaminated streams in central Texas was conducted to assess thyroid endocrine disruption. A survey of adult male and female cricket frogs (Acris crepitans) was performed at the same site between 2001 and 2003. Perchlorate is an oxidizer primarily used in solid-fuel rockets, and many sites that processed or used perchlorate are now contaminated. Histological analysis revealed that the fish from contaminated sites had increased thyroid follicular hyperplasia, hypertrophy, and colloid depletion. Multivariate analysis was generally found to be more powerful than univariate analysis. Seasonal differences existed in the degree of thyroidal perturbation were discovered, and fish were generally less sensitive to thyroidal perturbations in March compared to October. Thyroidal histological indicators were also correlated to levels of perchlorate in the fish, water, and periphyton. Periphyton was frequently most strongly correlated to thyroidal indices, suggesting that exposure through the food chain may be of import. In addition, one of the presumed reference sites turned out to be contaminated with perchlorate, and this was reflected by thyroidal biomarkers before perchlorate was detected in the stream water or biota. There was no evidence of colloid depletion or hyperplasia in frogs from any of the sites, although frogs from two sites with greatest mean water perchlorate concentrations exhibited significantly greater follicle cell hypertrophy. Furthermore, there was a significant positive correlation between follicle cell height and mean water perchlorate concentrations for frogs collected from all sites. This is the first known published account of perchlorate-induced thyroid disruption in fish under field situations, only the second known published account for amphibians, and also points out the value of biomarkers for contaminant biomonitoring. PMID- 16341612 TI - What does the WHOQOL-Bref measure? Measurement overlap between quality of life and depressive symptomatology in chronic somatoform pain disorder. AB - INTRODUCTION: Quality of life has become an important outcome criterion for psychiatric interventions. Especially in chronic disorders with no complete recovery, the improvement of quality of life is an important treatment goal. Nevertheless, there are methodological problems in assessing quality of life. There is a possible measurement overlap between quality of life and psychopathology, especially depression, which may invalidate research results. This study addresses the quality of life of patients with chronic somatoform pain and its relation to depressive symptoms. METHOD: One hundred out-patients with somatoform pain disorder at the Behavioural Medicine Pain Clinic in the Department of Psychiatry of the Medical University of Vienna were diagnosed using the SCID for Diagnostic and Statistical Manual of Mental Disorder (DSM-IV). The patients filled out the World Health Organisation Quality of Life Assessment-Bref (WHOQOL-Bref) and the Beck Depression Inventory (BDI). Pain intensity (average pain, maximum pain, minimal pain) and disability (work, leisure and family life) were assessed using visual analogue scales. RESULTS: Quality of life in somatoform pain disorder was reduced compared to the norm population, especially in the physical and psychological domains. There were highly significant negative correlations between, on the one hand, depressive symptomatology (BDI) and, on the other hand, the physical quality of life domain (r=-0.655, p<0.01), the psychological domain (r=-0.735, p<0.01), the social domain (r=-0.511, p<0.01) and the environmental domain (r=-0.561, p<0.01). In all domains of the WHOQOL-Bref and in the global score, significant differences between the group of patients with severe or very severe depressive symptoms and the group with no or only mild depressive symptoms were found. DISCUSSION: While the WHOQOL-Bref showed a poor quality of life of patients with chronic somatoform pain disorder in general and especially in the physical and in the psychological domains, the high correlation of physical and psychological quality of life scores with depressive symptomatology points to a measurement overlap. It is suggested that assessment of subjective quality of life should always be checked for the influence of depressive symptomatology on the quality of life score. PMID- 16341613 TI - Eating disorders. Prevalence and risk profile among secondary school students. AB - BACKGROUND: Eating disorders (EDs) are an important public health problem in developed countries. Despite the amount of epidemiological studies and causal theories, there is a great disparity of estimates and many questions remain still unclear. The aim of this study was to estimate the prevalence of the population at risk of developing EDs and describe the risk profiles among adolescents and young people. METHODS: A transversal design using a two-stage cluster sample of secondary education students aged between 12 and 18 years was used. The survey consisted of a socio-demographic questionnaire along with the Eating Attitudes Test (EAT)-40 index and was carried out between May and June 1999. RESULTS: Results showed that 7.8% of the secondary school population had a high risk of developing eating disorders (EAT-40 higher than 30). Females presented a higher percentage [12.3%; 95% confidence limits (CL) 10.4-14.3%] than males (3.2%; 95% CL 2.1-4.2%) at the highest risk levels. A decreasing risk gradient was observed with age. CONCLUSIONS: The estimates show an important difference between the risk in females and in males and suggests that the risk is incremented above all in the last few years of infancy and the first years of adolescence, when these boys and girls are more vulnerable to socio-cultural influences and do not yet have defined image and "behaviour models". PMID- 16341614 TI - North Birmingham assertive outreach evaluation. Patient characteristics and clinical outcomes. AB - BACKGROUND: Given the adoption of assertive outreach teams into UK mental health policy, it is important to assess whether gains identified in efficacy studies can be replicated in ordinary clinical settings. AIM: The aim of the study was to assess patient characteristics and clinical outcomes in routine assertive outreach services in the UK. METHODS: Patients (N=250), newly taken onto five assertive outreach teams, were followed up over 2 years. Baseline characteristics and outcome measures were compared. RESULTS: Most patients had an International Statistical Classification of Diseases and Related Health Problems, 10th Revision, diagnosis of schizophrenia, schizotypal or delusional disorder and long standing involvement with psychiatric services. Around a half had a history of substance abuse and violence. At follow-up, the majority had ongoing input from the teams and there was a significant decrease in the use of in-patient care. There was no improvement in symptoms, risk behaviours or social functioning. CONCLUSION: Patients on assertive outreach teams remain in contact with services and spend less time in hospital yet show little change with respect to clinical outcomes. PMID- 16341615 TI - Predicting internalizing problems in Moroccan immigrant adolescents in The Netherlands. AB - BACKGROUND: With the increasing number of immigrants worldwide, it is essential to have insight into the factors associated with internalizing problems in immigrant youth. However, little research on this subject has been conducted. The aim of the current study is to contribute to the knowledge in this field. METHODS: Data were obtained from the general population of 11- to 18-year-old Moroccan immigrant adolescents in The Netherlands. Using the Child Behavior Checklist (CBCL), Youth Self-Report (YSR), and Teacher's Report Form (TRF), 415 parent, 376 self-, and 238 teacher reports were available for analysis. RESULTS: The data showed relations between internalizing problems and several child (externalizing and chronic health problems), proximal family (paternal and maternal support and parent-child conflict), contextual family (conflicts between parents about parenting and total number of life-events), school/peer (being bored), and migration variables (adolescent's perceived discrimination). Moreover, a modest relation was found between internalizing problems and parental psychopathology. Few associations occurred with the global family factor (e.g., family educational level). Several relations between the predictors and YSR internalizing proved to be gender-specific. CONCLUSIONS: Our results suggested that the child, school/peer, and proximal family factors are the most important in predicting the development of internalizing problems in Moroccan immigrant adolescents. The impact of the migration factor was small. PMID- 16341616 TI - Alcohol dependence, excessive drinking and deliberate self-harm: trends and patterns in Oxford, 1989-2002. AB - BACKGROUND: Problems relating to alcohol use are very common among deliberate self-harm (DSH) patients, and alcohol abuse increases the risk of both DSH and suicide. In the UK, per capita consumption of alcohol has risen by 50% since 1970. The proportion of women (but not men) drinking in excess of government recommended limits has also increased. We investigate trends, by gender and age group, in alcohol problems and usage among DSH patients. METHOD: Data collected by the Oxford Monitoring System for Attempted Suicide were used to examine trends in alcohol disorders and alcohol consumption shortly before, or at, the time of self-harm by patients aged 15 years or over between 1989 and 2002. RESULTS: Data were available on 10,414 patients who were involved in 17,511 episodes of DSH. The annual numbers of both male and female DSH patients rose progressively over the study period. Although rates of alcohol disorders and consumption remained higher in males than females, substantial increases were seen in females of all ages in rates of alcohol problems, excessive drinking and consumption of alcohol within 6 h of DSH and as part of the act of DSH. Rates for males largely remained unchanged. CONCLUSIONS: There has been a significant increase in excessive drinking and consumption of alcohol around the time of DSH by females but not males. These changes may relate to increases in the affordability and availability of alcohol and to social changes in drinking patterns. They have implications for services for DSH patients and may have an impact on future patterns of suicidal behaviour. PMID- 16341617 TI - Is terror gender-blind? Gender differences in reaction to terror events. AB - OBJECTIVE: This study examines gender differences in posttraumatic vulnerability in the face of the terror attacks that occurred during the Al-Aqsa Intifada. In addition, the contribution of level of exposure, sense of safety, self-efficacy, and coping strategies is assessed. METHOD: Participants were 250 men and 262 women, who constitute a representative sample of Israel's adult population. Data were collected via a structured questionnaire consisting of 51 items that were drawn from several questionnaires widely used in the study of trauma. RESULTS: The findings indicate that women endorsed posttraumatic and depressive symptoms more than men and that, generally, their odds of developing posttraumatic stress symptoms are six times higher than those of men. Results also revealed that women's sense of safety and self-efficacy are lower than men's and that there are gender differences in coping strategies in the face of terror. CONCLUSIONS: Gender differences in vulnerability to terror may be attributable to a number of factors, among these are women's higher sense of threat and lower self-efficacy, as well as their tendency to use less effective coping strategies than men. Level of exposure to terror was ruled out as a possible explanation for the gender differences in vulnerability. PMID- 16341618 TI - Assessing the construct validity of three indicators of psychological distress in relation to perceived health and physical illness. AB - OBJECTIVES: The aim of this study is to compare three indicators of psychological distress (PD) on the strength of their association with subjective (or perceived) health and to analyse to what extent these associations will change after adjusting for physical illness measures and other possible confounding variables. METHODS: Data were used from a community-based sample of adults (N=9,428). Psychological distress was measured using three different instruments: the Negative Affect Scale of Bradburn, a nervousness scale, and a self-reported depressive complaints. Physical illness was measured by seven specific chronic conditions, a co-morbidity index of 17 conditions and two disability measures. Subjective health was assessed by a single question. Ordinary least square and logistic regression as well as structural equation modelling were used to analyse the data. RESULTS: The relation between subjective health and PD is strongest in case nervousness and this, or negative affect, are used as indicators of PD. The measure of depressive complaints is less strongly, but still substantially, related to subjective health. After correction for physical illness variables, the change in strength of the association is slightest for depressive complaints and highest for nervousness. Only small differences between negative affect and nervousness were established. These measures, which were more contaminated by physical ill health than depressive complaints, have the strongest association with subjective health both before as well as after correction for physical illness components. CONCLUSION: Negative affect and nervousness are reliable and valid indicators of PD, which can be used to predict subjective health. However, for this purpose, a correction for the confounding effects of physical illness variables will be necessary. The depressive complaints measure is not only less predictive of subjective health but also less contaminated by physical illness variables, making it a better indicator of PD if correction for physical illness variables is not possible. PMID- 16341619 TI - Physical and mental health of Afghan, Iranian and Somali asylum seekers and refugees living in the Netherlands. AB - CONTEXT: Worldwide, the number of refugees and asylum seekers is estimated to be about 11.5 million plus a much larger number of former refugees who have obtained a residence permit in a new country. Although asylum seekers have been coming to the Netherlands since the 1980s, very few epidemiological studies have focused on this group of inhabitants or on the refugees who have resettled in this country. OBJECTIVES: The objectives of this study were to estimate the prevalence rates of physical and mental health problems and to identify the risk factors for these complaints. DESIGN, SETTING, AND PARTICIPANTS: A population-based study was conducted in the Netherlands from June 2003 to April 2004 among adult refugees and asylum seekers from Afghanistan, Iran and Somalia. Asylum seekers were living in 14 randomly selected reception centres, and random samples of refugees were obtained from the population registers of three municipalities (Arnhem, Leiden and Zaanstad). A total of 178 refugees and 232 asylum seekers participated (response rates of 59 and 89%, respectively). MAIN OUTCOME MEASURES: General health and physical health were measured with the Short-Form 36 and a list of 19 chronic conditions, respectively; symptoms of post-traumatic stress disorder (PTSD), depression and anxiety, were measured with the Harvard Trauma Questionnaire and the Hopkins Symptoms Checklist-25. RESULTS: More asylum seekers (59.1%) than refugees (42.0%) considered their health to be poor (P=0.001). In both groups, approximately half of the respondents suffered from more than one chronic condition. More asylum seekers than refugees had symptoms of PTSD (28.1 and 10.6%, respectively; P=0.000) and depression/anxiety (68.1 and 39.4, respectively; P=0.000). Respondents from Afghanistan and, in particular, from Iran had a higher risk for PTSD and depression/anxiety. Female gender was associated with chronic conditions, PTSD and depression/anxiety, and higher age was associated with poor general health and chronic conditions. A greater number of traumatic events was associated with all health outcomes, and more post migration stress and less social support were associated with PTSD and depression/anxiety symptoms. CONCLUSIONS: Both physical and mental health problems are highly prevalent among refugees and asylum seekers in the Netherlands. Although higher prevalence rates for most health outcomes were found among asylum seekers, both the specific health services for asylum seekers and the general health services in the municipalities should be aware of these problems. PMID- 16341620 TI - Secondary traumatization among wives of Israeli POWs: the role of POWs' distress. AB - BACKGROUND: The aim of the study is to examine secondary traumatization of wives of former prisoners of war (POWs) as manifested in posttraumatic stress disorder (PTSD) symptoms, additional psychiatric symptoms, and marital adjustment. In addition, it assessed the role of several contributors to the wives' secondary traumatization: the husband's PTSD, the level of his verbal and physical aggression, and the wife's level of self-disclosure. METHODS: The study compared three groups of Israeli wives: wives of POWs with PTSD (N=18), wives of POWs without PTSD, (N=64), and a control group of wives of veterans without PTSD (N=72). RESULTS: The highest level of distress in all measures was endorsed by the wives of POWs with PTSD. Moreover, in addition to husband's PTSD and captivity, both the man's aggression and the wife's self-disclosure played a role in the wife's level of distress. CONCLUSIONS: The findings show that the husbands' PTSD was more strongly associated with the wives' secondary traumatization than their captivity. PMID- 16341621 TI - Massive prolapse of the urethral mucosa following periurethral injection of calcium hydroxylapatite for stress urinary incontinence. AB - The injection of bulking agents into the urethral submucosa is designed to create artificial urethral cushions that can improve urethral coaptation and hence restore continence. Ideally, a urethral bulking agent should be non-immunogenic and biocompatible, leading to minimal inflammatory and fibrotic response. The authors present a case report of a granulomatous reaction leading to urethral prolapse, 3 months after the transurethral injection of calcium hydroxylapatite. To our knowledge, this is the first granulomatous reaction described after calcium hydroxylapatite injection. PMID- 16341623 TI - Determinants of health-related quality of life in women with vertebral fractures. AB - Health-related quality of life (HRQL) is an important consideration in the management of patients with vertebral fractures. The purpose of this study was to examine patient-related factors that contribute to HRQL after vertebral fractures, including co-morbidities, medications, fracture history, family disease history, demographics, exercise, education and living environment. A total of 1,129 post-menopausal women (mean age 67.2, SD 11.9 years) was studied from the Canadian Database of Osteoporosis and Osteopenia (CANDOO). HRQL was measured using the mini-osteoporosis quality of life questionnaire (mini-OQLQ). Separate multivariable linear regression analyses [parameter estimates and corresponding 95% confidence intervals (CI)] were performed for each of the five mini-OQLQ domains: symptoms, physical functioning, emotional functioning, activities of daily living and leisure domains. A strong positive association was found between HRQL and post-secondary education, a family history of osteoporosis, working and thiazide therapy. Exercise improved HRQL; however, several hours a week were required to be meaningful. Living in long-term care had the most marked negative effect on HRQL. Smoking, past surgery of the hip or spine, sedatives, anticonvulsants, atherosclerotic disease and hypertension were also associated with a substantially decreased HRQL across several domains. Calcium channel-blockers, chemotherapy, corticosteroids, diabetes, migraines, the number of non-vertebral fractures and falls had a negative impact on selected domains. We demonstrated that several modifiable factors influence HRQL in patients with vertebral fractures, and physicians should be aware of these and other markers of reduced HRQL to enhance patient care. PMID- 16341624 TI - Laparoscopic or open preperitoneal repair in the management of recurrent groin hernias. AB - Laparoscopic repair is considered by many to be the operation of choice for a recurrent hernia. The aim of this study was to compare long-term outcome of laparoscopic and open preperitoneal repair of recurrent groin hernias. All patients operated on by one surgeon for recurrent groin hernia between January 1994 and December 2001 were reviewed. Forty-five percent had their data collected prospectively, while in 55% the data was collected retrospectively. Over the study period 128 patients underwent repair of a recurrent groin hernia of whom 99 had either a laparoscopic or open preperitoneal repair. The mean age was 60 years (range 15-88), 93 were men while 6 were women. Forty-five had their hernia repaired laparoscopically while 54 had an open preperitoneal repair. Patients have been followed for a median of 5 years (range 2-9 years). There has been no recurrence in either group of patients. Two patients in the laparoscopic group and four in the open group suffered from chronic groin pain. One patient in either group developed a Hydrocele that was dealt with surgically. Laparoscopic and open preperitoneal repair of a recurrent groin hernia are associated with similar long-term outcomes. PMID- 16341622 TI - The role of collagen in bone strength. AB - Bone is a complex tissue of which the principal function is to resist mechanical forces and fractures. Bone strength depends not only on the quantity of bone tissue but also on the quality, which is characterized by the geometry and the shape of bones, the microarchitecture of the trabecular bones, the turnover, the mineral, and the collagen. Different determinants of bone quality are interrelated, especially the mineral and collagen, and analysis of their specific roles in bone strength is difficult. This review describes the interactions of type I collagen with the mineral and the contribution of the orientations of the collagen fibers when the bone is submitted to mechanical forces. Different processes of maturation of collagen occur in bone, which can result either from enzymatic or nonenzymatic processes. The enzymatic process involves activation of lysyl oxidase, which leads to the formation of immature and mature crosslinks that stabilize the collagen fibrils. Two type of nonenzymatic process are described in type I collagen: the formation of advanced glycation end products due to the accumulation of reducible sugars in bone tissue, and the process of racemization and isomerization in the telopeptide of the collagen. These modifications of collagen are age-related and may impair the mechanical properties of bone. To illustrate the role of the crosslinking process of collagen in bone strength, clinical disorders associated with bone collagen abnormalities and bone fragility, such as osteogenesis imperfecta and osteoporosis, are described. PMID- 16341626 TI - Is herniography useful? AB - Herniography has been used for 25 years in the diagnosis of occult herniation but has not gained widespread acceptance in the UK, despite studies confirming its high sensitivity and specificity for occult hernias and an excellent record of safety and patient acceptability. The traditional approach in the UK to suspected occult groin herniation has been surgical exploration. This study examined the use of herniography in a single district general hospital to assess its impact in limiting unnecessary groin explorations and allowing discharge of patients without hernias. The case notes of 90 successive patients referred for herniography by the department of general surgery in a single UK district general hospital over an 18-month period were reviewed. Eighty-seven completed examinations were analysed in which 23 hernias were diagnosed in 20 patients. Thirteen patients have undergone hernia repair with resolution of symptoms. There were no false positive examinations, although two inguinal hernias were incorrectly diagnosed radiologically as femoral hernias; there were two false negative examinations where additional hernias were found at laparoscopic repair. There were no reported complications. Twenty-four patients were discharged directly from the surgical clinic after a negative herniogram. Thirty patients were referred to other specialities. No patient had undergone groin exploration after a negative herniogram. Herniography is a useful tool in assessing obscure groin pain and potential occult herniation. It can reliably rule out the presence of a hernia and avoid the need for surgical exploration. Many patients with a negative herniogram can be reassured and discharged, whilst others may be referred on to other specialities safe in the knowledge that an occult hernia has been excluded. PMID- 16341627 TI - Physiological repair of inguinal hernia: a new technique (study of 860 patients). AB - The author has developed a new operation technique based on the physiological principle that provides dynamic posterior wall for inguinal hernia repair. Results of the first series of 400 patients were published in 2001 (ANZ J Surg 71:241-244, 2001). Now the author has described the results of the second series of 860 patients having 920 hernias with follow-up for more than 7 years. An undetached strip of the external oblique aponeurosis (EOA) is sutured to the inguinal ligament below and the muscle arch above, behind the cord, to form a new posterior wall. External oblique muscle gives additional strength to the weakened muscle arch to keep this strip physiologically dynamic. In this prospective study, 920 inguinal hernia repairs were performed between August 1990 and December 2003 in 860 patients. Follow-up was done for 7 years. The main outcome measure was early and late morbidities and especially recurrence in a long-term follow-up. Mean patient age was 50.5 years (range 18-90). A total of 851 (98.95%) patients were operated under local or regional anesthesia; 838 (97.4%) patients were ambulatory with limited movements in 6 h and free movements in 18-24 h; 792 (92%) patients had a hospital stay of one night and 840 (97.6%) patients returned to normal activities within 1-2 weeks. Hematoma formation requiring drainage was observed in one patient, while seven patients had wound edema during the postoperative period which subsided on its own. Follow-up was completed in 623 patients (72.5 %) by clinical examination or questionnaire. The median follow-up period was 7.8 years (range 1-12 years). There was no recurrence of hernia or postoperative neuralgia. This operation is simple to perform, does not require foreign body like a mesh or complicated dissection of the inguinal floor as in Bassini/Shouldice. It has shown excellent results with virtually zero recurrence rates. PMID- 16341628 TI - Xanthogranuloma of the sellar region. AB - The incidence of diagnosed xanthogranuloma of the sellar region is very low [1, 2, 5, 6]. We report about two cases 1) in a 57-year-old female and 2) in a 5-year old boy. In both cases radiographic findings revealed an inhomogeneous, contrast enhancing sellar lesion. Histopathology showed the typical features of a xanthogranuloma of the sellar region with cholesterol clefts, lympho plasmacellular infiltrates, marked hemosiderin deposits, multinucleated foreign body giant cells around cholesterol clefts, accumulation of macrophages and only small epithelial cell clusters [6]. As xanthogranuloma of the sellar region are rarely diagnosed we want to draw attention to this rather unusual diagnosis. PMID- 16341629 TI - Acute spontaneous subdural haematoma after transsphenoidal surgery. AB - Only one case of acute spontaneous subdural haematoma (ASSDH) has been reported following transsphenoidal surgery, whereas, another case of chronic SDH has been reported. We present a patient with pituitary apoplexy, who was treated by transsphenoidal surgery. 8 days postoperative he developed an ASSDH requiring immediate surgical evacuation. This case documents the third occurrence of SDH as a complication of transsphenoidal surgery and is the first report of ASSDH that required immediate surgical evacuation. PMID- 16341630 TI - Intramedullary clear cell meningioma. AB - Intramedullary clear cell meningioma (CCM), which is more aggressive than other meningioma variants, is extremely rare. To date, only one case of such a spinal tumour has been documented. We report the first case of an intramedullary CCM originating in the thoracic region of the spinal cord. PMID- 16341631 TI - Lesions identified on T2*-weighted gradient echo images in two patients with suspected diffuse axonal injury that resolved in less than ten days. AB - T2*-weighted gradient echo (GE) imaging is useful for detection of intracranial hemorrhage in the patients with diffusion axonal injury (DAI). However, the temporal changes in the DAI-related lesions on T2*-weighted GE images are not clear. We report two very rare cases with DAI in which lesions identified on T2* weighted GE images resolved in less than ten days. PMID- 16341632 TI - Predictive factors for arm pain, neck pain, neck specific disability and health after anterior cervical decompression and fusion. AB - BACKGROUND: Predictive factors for a low arm and neck pain, and good health after anterior cervical decompression and fusion (ACDF) with a cervical carbon fibre intervertebral fusion cage (CIFC) are still lacking. METHOD: A prospective consecutive study to investigate which preoperative factors that could predict a good outcome with regard to arm pain, neck pain, Neck Disability Index (NDI) and general health three years after ACDF with CIFC was conducted. Thirty-four patients were included before surgery. Measurements took place the day before, six months, one year and three years after ACDF. FINDINGS: In multivariate analysis, to be a non-smoker before surgery was the most important factor for a low postoperative arm pain, a low pain frequency was the most important factor for low postoperative neck pain, normal rating on Distress and Risk Assessment Method (DRAM) was the most important factor for high function on NDI and a low initial pain intensity was the most important factor for good postoperative health. For all outcome variables a normal rating on DRAM was an important factor for a good outcome. CONCLUSIONS: Non-smoking, a low pain level and normal rating on DRAM were the best preoperative predictors of a good outcome in ACDF. Inclusion criteria for surgery should be based on a bio psychosocial model and DRAM seems to be useful for including the traditional inclusion criteria. PMID- 16341633 TI - Anterior cervical fusion with polyetheretherketone (PEEK) cages in the treatment of degenerative disc disease. Preliminary observations in 36 consecutive cases with a minimum 12-month follow-up. AB - STUDY DESIGN: Retrospective analysis of 36 cases of degenerative disc disease treated by interbody fusion with polyetheretherketone (PEEK) cages. OBJECTIVE: To determine the safety and efficiency of PEEK cages for anterior cervical fusion (ACF). SUMMARY OF BACKGROUND DATA: ACF with autologous bone has been reported since over 50 years ago. The recent development of cages housing materials inducing osteogenesis simplifies the technique of interbody fusion. The main purposes of bone substitutes for ACF are immediate biomechanical support, osteo integration of the graft, and elimination of local side effects at the donor site. This report shows our results using PEEK cages. MATERIALS AND METHODS: During an 18-month period, 36 consecutive patients had cervical fusions at 43 levels between C3 and C7. All operations involved one or two disc spaces for degenerative disc disease. We implanted all disc spaces with PEEK cages (Stryker Corporation, Kalamazoo, MI) containing granulated coralline hydroxylapatite (Pro Osteon 200, Interpore Cross International, Irvine, CA) or deantigenated pig bone in a gel solution (Gen-Os, Tecnoss, Torino, Italy). RESULTS: About 97% of patients had a good to excellent outcome; the result in one myelopathic patient was fair. The cervical fusion rate was 16.7% at 3 months, 61.1% at 6 months, and 100% at one year. CONCLUSIONS: PEEK cages appear to be safe and efficient for ACF. In order to confirm our preliminary impressions studies on larger series with long term follow-up are warranted. PMID- 16341634 TI - Enhanced visualization methods for computed radiography images. AB - This paper focuses on the application of two image enhancement techniques for the picture archiving and communications systems imaging diagnostic workstation applied to computed radiography (CR) and digital radiography images. The first method is a contrast enhancement technique based on a class of nonlinear intensity transformations described by analytic transfer functions derived from Hurter and Driffield characteristic curves. The second method employs antialiasing techniques preventing the formation of Moire patterns on subsampled CR images containing antiscatter grid lines, designed to achieve a good balance between artifact suppression and resolution degradation. These techniques are likely to become standard features for all high-end medical imaging workstations in the near future, and thus, we are suggesting that more powerful descriptions of these types of processing should be included in the Digital Imaging and Communications in Medicine standard. PMID- 16341635 TI - Impact of PACS on dictation turnaround time and productivity. AB - This study was conducted to measure the impact of PACS on dictation turnaround time and productivity. The radiology information system (RIS) database was interrogated to calculate the time interval between image production and dictation for every exam performed during three 90-day periods (the 3 months preceding PACS implementation, the 3 months immediately following PACS deployment, and a 3-month period 1 year after PACS implementation). Data were obtained for three exam types: chest radiographs, abdominal CT, and spine MRI. The mean dictation turnaround times obtained during the different pre- and post PACS periods were compared using analysis of variance (ANOVA). Productivity was also determined for each period and for each exam type, and was expressed as the number of studies interpreted per full-time equivalent (FTE) radiologist. In the immediate post-PACS period, dictation turnaround time decreased 20% (p < 0.001) for radiography, but increased 13% (ns) for CT and 28% (p < 0.001) for MRI. One year after PACS was implemented, dictation turnaround time decreased 45% (p < 0.001) for radiography and 36% (p < 0.001) for MRI. For CT, 1 year post-PACS, turnaround times returned to pre-PACS levels. Productivity in the immediate post PACS period increased 3% and 38% for radiography and CT, respectively, whereas a 6% decrease was observed for MRI. One year after implementation, productivity increased 27%, 98%, and 19% in radiography, CT, and MRI, respectively. PACS benefits, namely, shortened dictation turnaround time and increased productivity, are evident 1 year after PACS implementation. In the immediate post-PACS period, results vary with the different imaging modalities. PMID- 16341636 TI - Impact of non-Legionella bacteria on the uptake and intracellular replication of Legionella pneumophila in Acanthamoeba castellanii and Naegleria lovaniensis. AB - In aquatic environments, Legionella pneumophila survives, in association with other bacteria, within biofilms by multiplying in free-living amoebae. The precise mechanisms underlying several aspects of the uptake and intracellular replication of L. pneumophila in amoebae, especially in the presence of other bacteria, remain unknown. In the present study, we examined the competitive effect of selected non-Legionella bacteria (Escherichia coli, Aeromonas hydrophila, Flavobacterium breve, and Pseudomonas aeruginosa) on the uptake of L. pneumophila serogroup 1 by the amoebae Acanthamoeba castellanii and Naegleria lovaniensis. We also investigated their possible influence on the intracellular replication of L. pneumophila in both amoeba species. Our results showed that the non-Legionella bacteria did not compete with L. pneumophila for uptake, suggesting that the amoeba hosts took in L. pneumophila through a specific and presumably highly efficient uptake mechanism. Living and heat-inactivated P. aeruginosa best supported the replication of L. pneumophila in N. lovaniensis and A. castellanii, respectively, whereas for both amoeba species, E. coli yielded the lowest number of replicated L. pneumophila. Furthermore, microscopic examination showed that 100% of the A. castellanii and only 2% of the N. lovaniensis population were infected with L. pneumophila at the end of the experiment. This study clearly shows the influence of some non-Legionella bacteria on the intracellular replication of L. pneumophila in A. castellanii and N. lovaniensis. It also demonstrates the different abilities of the two tested amoeba species to serve as a proper host for the replication and distribution of the human pathogen in man-made aquatic environments such as cooling towers, shower heads, and air conditioning systems with potential serious consequences for human health. PMID- 16341637 TI - Molecular characterization of an oil-degrading cyanobacterial consortium. AB - Recent studies have shown that the cyanobacterium Microcoleus chthonoplastes forms a consortium with heterotrophic bacteria present within the cyanobacterial sheath. These studies also show that this consortium is able to grow in the presence of crude oil, degrading aliphatic heterocyclic organo-sulfur compounds as well as alkylated monocyclic and polycyclic aromatic hydrocarbons. In this work, we characterize this oil-degrading consortium through the analysis of the 16S rRNA gene sequences. We performed the study in cultures of Microcoleus grown in mineral medium and in cultures of the cyanobacterium grown in mineral medium supplemented with crude oil. The results indicate that most of the clones found in the polluted culture correspond to well-known oil-degrading and nitrogen fixing microorganisms, and belong to different phylogenetic groups, such as the Alpha, Beta, and Gamma subclasses of Proteobacteria, and the Cytophaga/Flavobacteria/Bacteroides group. The control is dominated by one predominant organism (88% of the clones) closely affiliated to Pseudoxanthomonas mexicana (similarity of 99.8%). The presence of organisms closely related to well known nitrogen fixers such as Rhizobium and Agrobacterium suggests that at least some of the cyanobacteria-associated heterotrophic bacteria are responsible for nitrogen fixation and degradation of hydrocarbon compounds inside the polysaccharidic sheath, whereas Microcoleus provides a habitat and a source of oxygen and organic matter. PMID- 16341638 TI - Sediment resuspension effects on the benthic microbial loop in experimental microcosms. AB - Sediment resuspension induced by anthropogenic disturbance is becoming a major threat of marine coastal ecosystems worldwide. The effects of sediment resuspension on the pelagic domain and on macro- and meiobenthos are well documented in the literature, whereas the effects on the benthic microbial components are nearly neglected. We have investigated the effects of sediment resuspension at two different disturbance levels on benthic bacterial abundance, biomass, and activities and on heterotrophic nanobenthos abundance in experimental microcosms. The results of our experiments pointed out that, independently from the amount of involved energy, sediment resuspension determined a general decrease of all benthic microbial components. The main effects consisted of a decreased abundance of the metabolically active bacterial fraction and of heterotrophic nanobenthos abundance. However, the amount of energy involved in sediment resuspension had differential effects on the structure and functioning of the benthic microbial loop, but only in the short term (i.e., within 36 h). Sediment resuspension had a stimulatory effect on activities of surviving bacterial cells, which, in turn, resulted in increased sediment organic C turnover rates. We hypothesize that such an effect, enhancing nutrient availability, might have relevant consequences on the trophic state of coastal marine ecosystem. PMID- 16341639 TI - Bacterial community succession in natural river biofilm assemblages. AB - Temporal bacterial community changes in river biofilms were studied using 16S rRNA gene-based polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) followed by sequence analysis. Naturally occurring biofilms were sampled in 2001 during an undisturbed 7-month low-water period in the River Garonne (SW France). During the sampling period epilithic biomass exhibited a particular pattern: two 3-month periods of accumulation that resulted in two peaks in summer and fall, each at about 25 g ash-free dry mass per square meter. Bacterial community DGGE profiles differed between the summer and fall biomass peaks and shared only 30% common operational taxonomic units (OTUs), suggesting the influence of seasonal factors on these communities. During the second biomass accrual phase, bacterial richness and the appearance of new OTUs fitted a conceptual model of bacterial biofilm succession. During succession, five OTUs (corresponding to Dechloromonas sp., Nitrospira sp., and three different Spirosoma spp.) exhibited particular patterns and were present only during clearly defined successional stages, suggesting differences in life-history strategies for epilithic bacteria. Co-inertia analysis of DGGE banding patterns and physical-chemical data showed a significant relationship between community structure and environmental conditions suggesting that bacterial communities were mainly influenced by seasonal changes (temperature, light) and hydrodynamic stability. Within the periods of stability, analysis of environmental variables and community patterns showed the dominant influence of time and maturation on bacterial community structure. Thus, succession in these naturally occurring epilithic biofilm assemblages appears to occur through a combination of allogenic (seasonal) and autogenic changes. PMID- 16341640 TI - Effects of viruses and predators on prokaryotic community composition. AB - Dialysis bags were used to examine the impact of predation and viral lysis on prokaryotic community composition (PCC) over a 5-day experiment in the oligomesotrophic Lake Pavin (France). The impact of the different predator communities (protists and metazoans) of prokaryotes was estimated by water fractionation (<5 microm: treatment filtered on 5 microm, without ciliates and metazoans; UNF: unfiltered treatment with all planktonic communities). Enrichments of natural viruses (<1.2 microm: with a natural virus concentration; <1.2 mum V and VV: with enrichment leading to a double or triple concentration of viruses, respectively) were used to indirectly assess the control of virioplankton. Viral activity was estimated from the frequency of visibly infected cells (FVIC). PCC was determined by fluorescence in situ hybridization (FISH) and terminal restriction fragment length polymorphism (T-RFLP). In this study, PCC was affected by the eukaryote communities (especially flagellates), and viruses to a lesser extent. Cyanobacteria declined significantly during the experiment and were highly correlated with the FVIC. In addition, the 503-bp terminal restriction fragment (T-RF) disappeared in treatments with virus enrichments, suggesting possible viral-associated mortality processes, whereas the 506-bp T-RF was not affected in these treatments. On one hand, these results suggest a control of the PCC: first, by viral lysis of some dominant phylotypes and second, by interspecific competition between resistant strains for the uptake of substrates released by this lysis. The increase of Archaea may suggest that these cells benefit such resources. On the other hand, the disappearance and the stable proportion of some dominant phylotypes suggested a selection pressure due to the predatory activity on prokaryotes. In conclusion, prokaryotic abundance appears to be mainly controlled by flagellate protists, which also affected PCC, whereas viruses seemed to be essentially responsible for profound changes in PCC via direct and indirect actions. PMID- 16341641 TI - Isolation and characterization of polymeric galloyl-ester-degrading bacteria from a tannery discharge place. AB - The culturable bacteria colonizing the rhizosphere of plants growing in the area of discharge of a tannery effluent were characterized. Relative proportions of aerobic, denitrifying, and sulfate-reducing bacteria were determined in the rhizosphere of Typha latifolia, Canna indica, and Phragmites australis. Aerobic bacteria were observed to be the most abundant group in the rhizosphere, and plant type did not seem to influence the abundance of the bacterial types analyzed. To isolate bacteria able to degrade polyphenols used in the tannery industry, enrichments were conducted under different conditions. Bacterial cultures were enriched with individual polyphenols (tannins Tara, Quebracho, or Mimosa) or with an undefined mixture of tannins present in the tannery effluent as carbon source. Cultures enriched with the effluent or Tara tannin were able to degrade tannic acid. Six bacterial isolates purified from these mixed cultures were able to use tannic acid as a sole carbon source in axenic culture. On the basis of 16S ribosomal DNA sequence analysis, these isolates were closely related to organisms belonging to the taxa Serratia, Stenotrophomonas maltophilia, Klebsiella oxytoca, Herbaspirillum chlorophenolicum, and Pseudomonas putida. PMID- 16341642 TI - Contribution of sediment respiration to summer CO2 emission from low productive boreal and subarctic lakes. AB - We measured sediment production of carbon dioxide (CO(2)) and methane (CH(4)) and the net flux of CO(2) across the surfaces of 15 boreal and subarctic lakes of different humic contents. Sediment respiration measurements were made in situ under ambient light conditions. The flux of CO(2) between sediment and water varied between an uptake of 53 and an efflux of 182 mg C m(-2) day(-1) from the sediments. The mean respiration rate for sediments in contact with the upper mixed layer (SedR) was positively correlated to dissolved organic carbon (DOC) concentration in the water (r(2) = 0.61). The net flux of CO(2) across the lake surface [net ecosystem exchange (NEE)] was also closely correlated to DOC concentration in the upper mixed layer (r(2) = 0.73). The respiration in the water column was generally 10-fold higher per unit lake area compared to sediment respiration. Lakes with DOC concentrations <5.6 mg L(-1) had net consumption of CO(2) in the sediments, which we ascribe to benthic primary production. Only lakes with very low DOC concentrations were net autotrophic (<2.6 mg L(-1)) due to the dominance of dissolved allochthonous organic carbon in the water as an energy source for aquatic organisms. In addition to previous findings of allochthonous organic matter as an important driver of heterotrophic metabolism in the water column of lakes, this study suggests that sediment metabolism is also highly dependent on allochthonous carbon sources. PMID- 16341669 TI - Mice overexpressing hepatic Abcb11 rapidly develop cholesterol gallstones. AB - Cholelithiasis is a polygenic disease, although the genes responsible for gallstone formation have not yet been clearly identified. QTL analysis has identified the Lith 1 loci on mouse Chromosome 2, and the hepatic bile salt transporter Abcb11 maps to the Lith 1 locus. We have used recently developed TTR Abcb11 transgenic mice that overexpress Abcb11 to determine the effects of Abcb11 overexpression on cholesterol gallstone formation. TTR-Abcb11 and FVB/NJ strain control mice were fed a lithogenic or chow diet and cholesterol crystal and gallstone formation were measured. Biliary lipids in gallbladder bile and gene expression of canalicular lipid transporters were also analyzed. TTR-Abcb11 mice fed a lithogenic diet had an increased rate of cholesterol crystal and gallstone formation. This was associated with an increase in both the hydrophobic bile salt and cholesterol content of gallbladder bile. Expression of Abcb4, Abcg5, and Abcg8 did not change before gallstone formation. These data indicate that hepatic overexpression of Abcb11 increases the rate of cholesterol gallstone formation. This is likely because of increases in bile salt hydrophobicity but not because of alterations of other biliary lipid transporters. These findings strongly support Abcb11 as a Lith 1 gene. PMID- 16341670 TI - Genetic alterations at the Bpag1 locus in dt mice and their impact on transcript expression. AB - The dystonin/Bpag1 gene encodes several tissue-specific alternatively spliced transcripts that encode cytoskeletal binding proteins. These various isoforms are necessary for maintaining the structural integrity of epithelial, neural, and muscle tissues. Mutations in the dystonin/Bpag1 gene cause dystonia musculorum (dt), a hereditary neuropathy of the mouse characterized by the progressive degeneration of sensory neurons. Several dt mutant alleles exist, most of which have arisen through spontaneous mutations. In this article we demonstrate that the dt locus encodes 107 exons spanning 400 kb. The high frequency of occurrence of spontaneous dt mutants may therefore be a result of the large size of the gene. Analysis of genomic DNA from several dt spontaneous mutant alleles, dt(24J), dt(27J), dt(Alb), and dt(Frk), shows a deletion of the central portion of the gene in dt(Alb) but no large rearrangements or deletions in the other alleles. These other alleles likely have small deletions or rearrangements, or point mutations. To determine the impact of the known and unknown mutations on transcript levels, RT-PCR was performed to detect various coding regions of the dystonin/Bpag1 transcripts in brain and muscle from multiple dt alleles: dt(Tg4), dt(Alb), dt(24J), dt(27J), and dt(Frk). With the exception of dt(Frk), reduced transcript levels were observed for all alleles tested. Such alterations likely result in reduced or absent dystonin/Bpag1 protein levels. Thus, distinct genetic defects lead to a common outcome of reduced transcript expression causing the same phenotype in multiple dt alleles. PMID- 16341671 TI - Mapping the dominant wound healing and soft tissue regeneration QTL in MRL x CAST. AB - We have used a mouse ear punch model and the QTL (quantitative trait loci) mapping technique to identify genes that are responsible for soft tissue regeneration. In the early studies, we have identified several QTL and have shown that the inheritance of ear healing was additive in one cross (MRL x SJL), and recessive in another cross (DBA x 129). Because CAST mice are genetically distinct and have a different genetic background, CAST would facilitate the identification of common and novel QTL when crossed with common inbred lines. We made a cross between super healer MRL and poor healer CAST and collected ear punch phenotype and marker genotype data from F(2). Ear punch healing exhibited a dominant mode of inheritance in this cross. There were three main QTL on Chromosomes 4, 9, and 17, and two suggestive QTL on Chromosomes 1 (new) and 7. Taken together, these QTL accounted for about 29% of total F2 variance of MRL x CAST. Compared with another study using the same cross, we found a totally different set of QTL. Two QTL interactions were identified by a full QTL model: Chromosomes 4 x 17 and 9 x 17; the latter reached to a statistical level at p < 0.05. These interactions explained about 4% of the F2 phenotypic variance. We conclude that soft tissue regeneration is controlled by multiple genes and locus vs. locus interactions. PMID- 16341672 TI - A catalog of nonsynonymous polymorphism on mouse chromosome 16. AB - Numerous phenotypic traits differ among inbred mice, and the genetic diversity of inbred strains has been exploited in studies of quantitative trait loci (QTL). Sequencing the mouse genome has resulted in improved tools for the study of QTL, but a comprehensive catalog of sequence variants between strains would be of great value in identifying and testing potentially causative alleles. A/J DNA was included in the Celera shotgun sequence of the mouse genome and C57BL/6 DNA was sequenced by an international consortium. We have resequenced A/J and B6 DNA to cover nearly all of the protein-coding portions of mouse Chromosome 16, revealing that there are 106 nonsynonymous substitutions in 74 of the 779 genes on the chromosome. The pattern of substitution is more similar to the spectrum of benign polymorphism in the human population than it is to human disease-causing mutations. In mouse, polymorphic variants tend to be associated with one another on large haplotypes; this pattern also holds true for nonsynonymous polymorphism. However, sufficient fragmentation of haplotypes is present to suggest that only a very-high-resolution haplotype map will enable effective inference of alleles in additional strains. PMID- 16341673 TI - Validation of mRNA/EST-based gene predictions in human Xp11.4 revealed differences to the organization of the orthologous mouse locus. AB - Careful manual annotation of the human reference sequence provides a solid basis for the identification of disease-associated genes. Toward this end, we focused on a medically relevant 2.6-Mb region of the human chromosome Xp11.4 between markers DXS9851 and DXS9751 and identified 16 transcription units according to the Vertebrate Genome Annotation (Vega) rules. In order to validate these annotations, we performed a comprehensive RT-PCR expression analysis and a human mouse comparison. This revealed, despite the high overall genomic conservation of the region, remarkable differences of the gene content between human and mouse. Whereas 12 of 16 annotations were confirmed by RT-PCR in human tissues, for only seven genes mouse orthologs could be identified and found to be expressed. This indicates that a comprehensive and experimentally supported annotation effort of the human genome simultaneously highlights regions with striking differences in gene organization to other species and may indicate evolutionary events specific to the human lineage demanding further functional analyses. PMID- 16341675 TI - Linkage analysis and gene expression profile of pancreatic acinar atrophy in the German Shepherd Dog. AB - Pancreatic acinar atrophy (PAA) is a degenerative disease of the exocrine pancreas and is the most common cause of exocrine pancreatic insufficiency in the German Shepherd Dog. Analyses of inheritance have shown that a single gene segregating in an autosomal recessive fashion is causative for PAA. To date the gene and causative mutation have not been determined. To identify a region of interest and/or candidate genes, we conducted linkage and gene expression studies. Analysis of 384 microsatellite markers resulted in a maximum two-point LOD score of 2.5 for FH2107 on CFA03. We used an oligonucleotide array to generate gene expression profiles for normal and affected pancreata. It revealed 244 genes with greater than two-fold difference in expression levels. Five genes of interest were further assessed by TaqMan quantitative real-time RT-PCR that confirmed trends observed using the microarray. One gene, gp25L, located on CFA03, was found to be downregulated by more than 500-fold in affected pancreata and was further investigated as a candidate gene. Sequence data did not reveal a mutation in the coding sequence that segregates with PAA. PMID- 16341676 TI - Prion-like Doppel gene (PRND) in the goat: genomic structure, cDNA, and polymorphisms. AB - The genomic structure of the caprine Doppel gene (PRND) was determined using the ovine sequence as a scaffold to generate PCR fragments that were aligned with a cDNA sequence obtained from testicular mRNA. The caprine gene contains two exons, 89 and >2291 bp long, separated by a 1689-bp intron. Two mRNA isoforms of 3.2 and 4.8 kb were identified in the testis, as well as the exact transcription start site by fluorescently labeled oligonucleotide extension (FLOE). Like in sheep and cattle, the open reading frame (ORF) (537 bp) lies within exon 2 and is very much conserved in sheep (99.3%) and cattle (97%). The intronic sequence is also highly conserved (95.3%) compared with sheep, with the only exception of a 47-bp insertion. The PRND ORF was sequenced in 47 healthy and 17 TSE-affected goats of the Italian Ionica breed. Seven nucleotide positions showed variation: T28C, C65T, A151G, G286A, C385G, T451C, and T528C. Five were commonly represented polymorphisms: T28C, T451C, and T528C are silent mutations at codons L10, L151, and I176, respectively, while A151G and C385G determine a T51A and L129V amino acid change, respectively. The two remaining variants, C65T and G286A, were rare, leading to the amino acid substitutions S22F and E96K, respectively. None of the polymorphisms was significantly relatable to the TSE status, and the same result was obtained by the analysis of the combined haplotypes at the five major polymorphic sites, namely, T28C, C65T, A151G, G286A, and C385G. PMID- 16341677 TI - [Modern pharmacotherapy of age-related macular degeneration]. AB - Age-related macular degeneration (AMD) is now the most common cause for blind registration in all developed countries. Epidemiologic data indicate that there are 4.5 millions affected in Germany with constant increase in incidence and prevalence with subsequent considerable health economic implications. Late manifestations of the disease result in the inability to read and to perform daily tasks. Therefore, there is an urgent need for efficacious prophylactic and therapeutic measures to prevent irreversible loss of central vision. Based on a better understanding of the underlying molecular mechanisms new therapeutic approaches have been brought forward and expand previous approaches such as thermal laser surgery or photodynamic therapy. Repeated intravitreal injection of anti-VEGF (vascular endothelial growth factor) agents as well as corticosteroids have a beneficial effect on growth and permeability of neovascular membranes. The risk for progression from early to late stages of AMD can be reduced with certain antioxidative preparations (AREDS medication) in presence of defined funduscopic signs. Early diagnosis is key for all currently available interventions since a beneficial effect can only be achieved in early stages of the disease process. PMID- 16341678 TI - [Endomicroscopy--technology with future]. AB - Endomicroscopy becomes possible due to the integration of a miniaturized confocal microscope in the distal tip of a conventional endoscope. Endomicroscopy enables subsurface analysis of the gut mucosa and in vivo histology during ongoing endoscopy in full resolution by point scanning laser fluorescence analysis. Cellular, vascular and connective structures can be seen in detail. Graduation of cellular changes with endomicroscopy allows an immediate in-vivo diagnosis of different gastrointestinal diseases. The diagnostic spectrum of confocal endomicroscopy is currently expanding from screening and surveillance for colorectal cancer towards Barrett's esophagus, Helicobacter pylori associated gastritis and early gastric cancer. The new detailed images seen with confocal laser endomicroscopy are unequivocally the beginning of a new era where this optical development will allow a unique look on cellular structures and functions at and below the surface of the gut. PMID- 16341679 TI - [Paraplegia after acute thoracic pain]. AB - Severe neurological complications such as spinal cord ischemia and paraplegia can occur with acute aortic dissection in 3%. This report describes the case of a 67 year old patient with delayed onset of paraplegia 8 h after acute chest pain. Contrast enhanced computed tomography documented Stanford type B dissection confined to a short segment of the aorta. Furthermore, magnetic resonance imaging revealed intraspinal intraaxial hematoma of the myelon, which can explain the neurological complication. This case shows that even in the scenario of acute aortic dissection other mechanisms for paraplegia may be operational than dissection itself. Paraplegia in this case results from intramyelon bleeding preceding aortic dissection. PMID- 16341674 TI - Transcriptome analysis of human gastric cancer. AB - To elucidate the genetic events associated with gastric cancer, 124,704 cDNA clones were collected from 37 human gastric cDNA libraries, including 20 full length enriched cDNA libraries of gastric cancer cell lines and tissues from Korean patients. An analysis of the collected ESTs revealed that 97,930 high quality ESTs coalesced into 13,001 clusters, of which 11,135 clusters (85.6%) were annotated to known ESTs. The analysis of the full-length cDNAs also revealed that 4862 clusters (51.7%) contained at least one putative full-length cDNA clone with an initiation codon, with the average length of the 5' UTR of 140 bp. A large number appear to have a diverse transcription start site (TSS). An examination of the TSS of some genes, such as TEGT and GAPD, using 5' RACE revealed that the predicted TSSs are actually found in human gastric cancer cells and that several TSSs differ depending on the specific gastric cell line. Furthermore, of the human gastric ESTs, 766 genes (9.5%) were present as putative alternatively spliced variants. Confirmation of the predicted spliced isoforms using RT-PCR showed that the predicted isoforms exist in gastric cancer cells and some isoforms coexist in gastric cell lines. These results provide potentially useful information for elucidating the molecular mechanisms associated with gastric oncogenesis. PMID- 16341680 TI - [Interdisciplinary treatment of carotid cavernous fistulas via the superior ophthalmic vein]. AB - BACKGROUND: The endovascular occlusion of symptomatic carotid cavernous fistulae (CCF) via the transfemoral approach is safe and effective. Due to anatomical variations or after unsuccessful transarterial therapy, a direct surgical approach to the superior ophthalmic vein (SOV) may be necessary. CASE REPORTS: In two patients with acute ophthalmologic symptoms coil occlusion of the CCF was performed after palpebral incision and cannulation of the SOV. RESULTS: In both patients preparation of the SOV was performed successfully and without complications. After coil embolization of the CCF both patients had complete resolution of symptoms within several weeks. During a follow-up of 12 months there were no recurrences, but both patients exhibited moderate blepharoptosis. CONCLUSION: Embolization of CCF via a surgically created approach is an effective procedure in selected cases when standard interventional treatment is not possible. PMID- 16341681 TI - Antibiotic management of ventilator-associated pneumonia due to antibiotic resistant gram-positive bacterial infection. AB - Gram-positive cocci, in particular Staphylococcus aureus, account for as much as one-third of all cases of hospital-acquired pneumonia, and treatment has become increasingly complex as the proportion of resistant isolates has increased. Methicillin-resistant S. aureus is of particular concern because this pathogen is now associated with hospital-acquired, ventilator-associated, community-acquired, and healthcare-associated pneumonia. Antibiotic therapy for ventilator-associated pneumonia is challenging because it can be caused by multiple pathogens, which can be resistant to multiple drugs. This article reviews the epidemiology of ventilator-associated pneumonia and describes options for antibiotic treatment. Particular attention is paid to pneumonia due to methicillin-resistant S. aureus. Studies suggest that vancomycin, the traditional treatment for ventilator associated pneumonia, may not be the best option for this type of pneumonia and that other antibiotics, such as linezolid and clindamycin, might be better choices. New antibiotics with activity against methicillin-resistant S. aureus are under investigation and may soon become available for clinical use. Studies are needed to define the optimal choice of antibiotic for pneumonias caused by this organism, and these choices will need to be balanced with the need to minimize the emergence of resistance. PMID- 16341682 TI - An additive-dominance model to determine chromosomal effects in chromosome substitution lines and other gemplasms. AB - When using chromosome substitution (CS) lines in a crop breeding improvement program, one needs to separate the effects of the substituted chromosome from the remaining chromosomes. This cannot be done with the traditional additive dominance (AD) model where CS lines, recurrent parent, and their hybrids are used. In this study, we develop a new genetic model and software, called a modified AD model with genotype x environment interactions, which can predict additive and dominance genetic effects attributed to a substituted alien chromosome in a CS line as well as the overall genetic effects of the non substituted chromosomes. In addition, this model will predict the additive and dominance effects of the same chromosome of interest (i.e. chromosome 25 of cotton in this study) in an inbred line, as well as the effects of the remaining chromosomes in the inbred line. The model requires a CS line, its recurrent parent and their F(1) and/or F(2) hybrids between the substitution lines and several inbred lines. Monte Carlo simulation results showed that genetic variance components were estimated with no or slight bias when we considered this modified AD model as random. The correlation coefficient between predicted effects and true effects due to the chromosomes of interest varied from zero to greater than 0.90 and it was positively relative to the difference between the CS line and the recurrent line. To illustrate the use of this new genetic model, an upland cotton, Gossypium hirsusum L, CS line (CS-B25), TM-1 (the recurrent parent), five elite cultivars, and the F(2) hybrids from test-crossing these two lines with the five elite cultivars were grown in two environments in Mississippi. Agronomic and fiber data were collected and analyzed. The results showed that the CS line, CS B25, which has chromosome 25 from line 3 to 79, Gossypium barbadense substituted into TM-1, had positive genetic associations with several fiber traits. We also determined that Chromosome 25 from FiberMax 966 had significantly positive associations with fiber length and strength; whereas, chromosome 25 from TM-1 and SureGrow 747 had detectable negative genetic effects on fiber strength. The new model will be useful to determine effects of the chromosomes of interest in various inbred lines in any diploid or amphidiploid crop for which CS lines are available. PMID- 16341683 TI - Development and QTL assessment of Triticum aestivum-Aegilops tauschii introgression lines. AB - A set of 84 bread wheat lines, each containing a single homozygous introgression of the Aegilops tauschii genome was produced in the 'Chinese Spring' background via backcrossing of the D-genome chromosome substitution lines 'Chinese Spring'/Sears's 'Synthetic 6x' with the recurrent parent and subsequent selfing. The development of the lines was accompanied by microsatellite marker assisted selection. With the exception of three telomeric regions at chromosomes 1DL, 4DL and 7DS, and a region of less than 24 cM on the chromosome arm 3DL, the genome of Ae. tauschii is fully represented in these lines. The newly developed lines were used for the discovery of morphological and agronomical quantitative trait loci (QTLs) from the wild species. Fifty-two introgression lines were grown in the field and evaluated for six traits including flowering time, plant height, ear length, spikelet number, fertility and grain weight per ear. Seventeen significant QTLs were detected, Ae. tauschii contributed favourable alleles at nine loci influencing five traits. The whole set of 84 homozygous lines provides a tool for further testing the effects and stability of the detected QTLs and for the evaluation of new traits. PMID- 16341684 TI - Characteristics, development and mapping of Gossypium hirsutum derived EST-SSRs in allotetraploid cotton. AB - In order to construct a saturated genetic map and facilitate marker-assisted selection (MAS) breeding, it is necessary to enhance the current reservoir of known molecular markers in Gossypium. Microsatellites or simple sequence repeats (SSRs) occur in expressed sequence tags (EST) in plants. Many ESTs are publicly available now and represent a good tool in developing EST-SSRs. From 13,505 ESTs developed from our two cotton fiber/ovule cDNA libraries constructed for Upland cotton, 966 (7.15%) contained one or more SSRs and from them, 489 EST-SSR primer pairs were developed. Among the EST-SSRs, 59.1% are trinucleotides, followed by dinucleotides (30%), tetranucleotides (6.4%), pentanucleotides (1.8%), and hexanucleotides (2.7%). AT/TA (18.4%) is the most frequent repeat, followed by CTT/GAA (5.3%), AG/TC (5.1%), AGA/TCT (4.9%), AGT/TCA (4.5%), and AAG/TTC (4.5%). One hundred and thirty EST-SSR loci were produced from 114 informative EST-SSR primer pairs, which generated polymorphism between our two mapping parents. Of these, 123 were integrated on our allotetraploid cotton genetic map, based on the cross [(TM-1xHai7124)TM-1]. EST-SSR markers were distributed over 20 chromosomes and 6 linkage groups in the map. These EST-SSR markers can be used in genetic mapping, identification of quantitative trait loci (QTLs), and comparative genomics studies of cotton. PMID- 16341686 TI - Regulation of insulin signalling, glucose uptake and metabolism in rat skeletal muscle cells upon prolonged exposure to resistin. AB - AIMS/HYPOTHESIS: Debate exists regarding the role of resistin in the pathophysiology of insulin resistance. The aim of this study was to directly assess the effects of resistin (0-24 h) on basal and insulin-stimulated glucose uptake and metabolism in skeletal muscle cells and to investigate the mechanisms responsible for the effects of resistin. METHODS: We used L6 rat skeletal muscle cells and examined [(3)H]2-deoxyglucose uptake, GLUT4 translocation and GLUT protein content. We assessed glucose metabolism by measuring the incorporation of D-[U-(14)C]glucose into glycogen, (14)CO(2) and lactate production, as well as the phosphorylation level and total protein content of insulin signalling proteins, including insulin receptor beta-subunit (IRbeta), insulin receptor substrate (IRS), Akt and glycogen synthase kinase-3beta (GSK-3beta). RESULTS: Treatment of L6 rat skeletal muscle cells with recombinant resistin (50 nmol/l, 0 24 h) reduced levels of basal and insulin-stimulated 2-deoxyglucose uptake and decreased insulin-stimulated GLUT4myc content at the cell surface, with no alteration in the production of GLUT4 or GLUT1. Resistin also decreased glycogen synthesis and GSK-3beta phosphorylation. Insulin-stimulated oxidation of glucose via the Krebs cycle was reduced by resistin, whereas lactate production was unaltered. Although insulin receptor protein level and phosphorylation were unaltered by resistin, production of IRS-1, but not IRS-2, was downregulated and a decreased tyrosine phosphorylation of IRS-1 was detected. Reduced phosphorylation of Akt on T308 and S473 was observed, while total Akt and Akt1, but not Akt2 or Akt3, production was decreased. CONCLUSIONS/INTERPRETATION: Our data show that resistin regulates the function of IRS-1 and Akt1 and decreases GLUT4 translocation and glucose uptake in response to insulin. Selective decreases in insulin-stimulated glucose metabolism via oxidation and conversion to glycogen were also induced by resistin. These observations highlight the potential role of resistin in the pathophysiology of type 2 diabetes in obesity. PMID- 16341685 TI - Prediction, progression and prevention of diabetic nephropathy. The Minkowski Lecture 2005. AB - Diabetic nephropathy is a major problem for patients and health care systems. The costs of treatment remain high. To confront the ongoing challenge, we need to identify individuals at high risk for initiation and progression of this devastating complication. Risk factors include genetic markers; constitutional factors such as low birthweight; haemodynamic factors, including activation of the RAS system and hypertension; metabolic factors such as glycaemia; and additional factors such as urinary AER and smoking. Modifiable risk factors should be treated aggressively. Potential new markers of risk include indices of increased inflammation, changes in coagulation, endothelial dysfunction, growth factors and cytokines. Application of such markers may in time improve risk assessment and allow new treatment targets to be identified. Interventions that aim to achieve strict glycaemic control and blockade of the renin-angiotensin system have been shown to be effective in clinical trials and are feasible in clinical practice. The 'natural history' of diabetic nephropathy can be transformed if these strategies of intensive screening and care are applied, leading both to a lower incidence of diabetic nephropathy and to an improved outcome, with survival exceeding 20 years from onset of overt proteinuria. PMID- 16341687 TI - Activator protein-1 in epiretinal membranes of patients with proliferative diabetic retinopathy. PMID- 16341688 TI - The impact of ethnicity on glucose regulation and the metabolic syndrome following gestational diabetes. AB - AIMS/HYPOTHESIS: We assessed the impact of ethnic origin on metabolism in women following gestational diabetes mellitus (GDM). MATERIALS AND METHODS: Glucose regulation and other features of the metabolic syndrome were studied at 20.0 (18.2-22.1) months (geometric mean [95% CI]) post-partum in women with previous GDM (185 European, 103 Asian-Indian, 80 African-Caribbean). They were compared with the same features in 482 normal control subjects who had normal glucose regulation during and following pregnancy. RESULTS: Impaired glucose regulation or diabetes by WHO criteria were present in 37% of women with previous GDM (diabetes in 17%), especially in those of African-Caribbean and Asian-Indian origin (50 and 44%, respectively vs 28% in European, p=0.009). BMI, waist circumference, diastolic blood pressure, fasting triglyceride and insulin levels, and insulin resistance by homeostatic model assessment (HOMA), were increased following GDM (p<0.001 for all, vs control subjects). Where glucose regulation was normal following GDM, basal insulin secretion (by HOMA) was high (p<0.001 vs control subjects). Irrespective of glucose regulation in pregnancy, Asian-Indian origin was associated with high triglyceride and low HDL cholesterol levels, and African-Caribbean with increased waist circumference, blood pressure, and insulin levels, together with insulin resistance and low triglyceride concentrations. Nonetheless, the GDM-associated features were consistent within each ethnic group. The metabolic syndrome by International Diabetes Federation criteria was present in 37% of women with previous GDM, especially in non-Europeans (Asian Indian 49%, African-Caribbean 43%, European 28%, p=0.001), and in 10% of controls. CONCLUSIONS/INTERPRETATION: Following GDM, abnormal glucose regulation and the metabolic syndrome are common, especially in non-European women, indicating a need for diabetes and cardiovascular disease prevention strategies. PMID- 16341689 TI - Do HbA1c levels and the self-monitoring of blood glucose levels adequately reflect glycaemic control during pregnancy in women with type 1 diabetes mellitus? AB - AIMS/HYPOTHESIS: Pregnancies of women with type 1 diabetes mellitus are associated with maternal and perinatal complications. These complication rates remain elevated despite achievement of the treatment goals described in the widely used guidelines of the American Diabetes Association (i.e. HbA(1)c level 7.0%. Self-monitored glucose levels and those measured with CGMS were compared between patients with four or five, six to nine and ten or more SMBG determinations daily. RESULTS: In patients with HbA(1)c levels 6.0%. In women with HbA(1)c levels 6.0-7.0% and >7.0%, these levels did not differ. The detection rate of hyper- and hypoglycaemic episodes was significantly higher in patients with ten or more SMBG determinations daily than in patients with fewer than ten. CONCLUSIONS/INTERPRETATION: Treatment of diabetes in pregnant women should be aimed at achieving HbA(1)c levels within the normal range, i.e. 3 months) were recruited. A 100 mm visual analogue scale (VAS) for perceived pain, the Roland Morris (RM) Questionnaire for low back function, and the SF-36 for overall health status served as primary outcome measures. Subjects were randomly allocated to receive either FD or ATEP. The FD intervention consisted of the application of flexion and traction applied to specific regions in the low back, with the aid of a specially designed manipulation table. The ATEP intervention included stabilizing and flexibility exercises, the use of modalities, and cardiovascular training. A total of 235 subjects met the inclusion/exclusion criteria and signed the informed consent. Of these, 123 were randomly allocated to FD and 112 to the ATEP. Study patients perceived significantly less pain and better function after intervention, regardless of which group they were allocated to (P<0.01). Subjects randomly allocated to the flexion-distraction group had significantly greater relief from pain than those allocated to the exercise program (P=0.01). Subgroup analysis indicated that subjects categorized as chronic, with moderate to severe symptoms, improved most with the flexion-distraction protocol. Subjects categorized with recurrent pain and moderate to severe symptoms improved most with the exercise program. Patients with radiculopathy did significantly better with FD. There were no significant differences between groups on the Roland Morris and SF-36 outcome measures. Overall, flexion-distraction provided more pain relief than active exercise; however, these results varied based on stratification of patients with and without radiculopathy and with and without recurrent symptoms. The subgroup analysis provides a possible explanation for contrasting results among randomized clinical trials of chronic low back pain treatments and these results also provide guidance for future work in the treatment of chronic low back pain. PMID- 16341713 TI - Neonatal intestinal obstruction simulating meconium ileus in infants with long segment intestinal aganglionosis: radiographic findings that prompt the need for rectal biopsy. AB - BACKGROUND: The initial clinical presentation and radiographic finding of microcolon in children with long-segment intestinal aganglionosis involving the entire colon, ileum and sometimes the jejunum can mimic meconium ileus. This makes the diagnosis difficult for the radiologist and surgeon. OBJECTIVE: To document and describe the clinical and radiographic findings in children with long-segment intestinal aganglionosis who are initially thought to have meconium ileus. MATERIALS AND METHODS: We reviewed the cases of six neonates with long segment intestinal aganglionosis presenting as meconium ileus at our institutions between 1978 and 2002. We examined the clinical presentation and the radiographic, surgical, and pathologic findings. In addition, 17 cases from the literature were identified and are included in the discussion. RESULTS: A total of 23 cases were reviewed. Right lower quadrant intraluminal calcifications were noted on abdominal radiographs in all six neonates of our series and were described in 13 of the 17 neonates reported in the literature. Similarly, a microcolon was present in five of the six neonates of our series and in 14 of 16 historical neonates (one not reported). CONCLUSION: In a neonate with small-bowel obstruction and a microcolon, the presence of right lower quadrant intraluminal calcifications should raise the suspicion of long-segment intestinal aganglionosis even if the operative findings are typical of meconium ileus and a biopsy should be performed. PMID- 16341714 TI - Estimating the timing of diet shifts using stable isotopes. AB - Stable isotope analysis has become an important tool in studies of trophic food webs and animal feeding patterns. When animals undergo rapid dietary shifts due to migration, metamorphosis, or other reasons, the isotopic composition of their tissues begins changing to reflect that of their diet. This can occur both as a result of growth and metabolic turnover of existing tissue. Tissues vary in their rate of isotopic change, with high turnover tissues such as liver changing rapidly, while relatively low turnover tissues such as bone change more slowly. A model is outlined that uses the varying isotopic changes in multiple tissues as a chemical clock to estimate the time elapsed since a diet shift, and the magnitude of the isotopic shift in the tissues at the new equilibrium. This model was tested using published results from controlled feeding experiments on a bird and a mammal. For the model to be effective, the tissues utilized must be sufficiently different in their turnover rates. The model did a reasonable job of estimating elapsed time and equilibrial isotopic changes, except when the time since the diet shift was less than a small fraction of the half-life of the slowest turnover tissue or greater than 5-10 half-lives of the slowest turnover tissue. Sensitivity analyses independently corroborated that model estimates became unstable at extremely short and long sample times due to the effect of random measurement error. Subject to some limitations, the model may be useful for studying the movement and behavior of animals changing isotopic environments, such as anadromous fish, migratory birds, animals undergoing metamorphosis, or animals changing diets because of shifts in food abundance or competitive interactions. PMID- 16341715 TI - Sexual reproduction, clonal diversity and genetic differentiation in patchily distributed populations of the temperate forest herb Paris quadrifolia (Trilliaceae). AB - Clonal plant species have been shown to adopt different strategies to persist in heterogeneous environments by changing relative investments in sexual reproduction and clonal propagation. As a result, clonal diversity and genetic variation may be different along environmental gradients. We examined the regional and local population structure of the clonal rhizomatous forest herb Paris quadrifolia in a complex of forest fragments in Voeren (Belgium). Relationships between population size (the number of shoots), shoot density (the number of shoots per m2) and local growth conditions were investigated for 47 populations. Clonal diversity and genetic variation within and among 19 populations were investigated using amplified fragment length polymorphism markers. To assess the importance of sexual reproduction, seed set, seed weight and germination success were determined in 18 populations. As predicted, local growth conditions largely affected population distribution, size and density of P. quadrifolia. Populations occurring in moist and relatively productive sites contained significantly more shoots. Here, shoots were also much more sparsely distributed compared to populations occurring in dry and relatively unproductive sites, where shoots showed a strongly aggregated distribution pattern. Clonal diversity was relatively high, compared with other clonal species (G/N ratio=0.43 and Simpson's D=0.81). Clonal diversity significantly (P<0.01) decreased with increasing shoot density while molecular genetic variation was significantly (P<0.01) affected by population size and local environmental conditions. Lack of recruitment and out-competition of less-adapted genotypes may explain the decreased genetic variation in dry sites. Analysis of molecular variance revealed significant genetic variation among populations (PhiST=0.42, P<0.001), whereas pairwise genetic distances were not correlated to geographic distances, suggesting that gene flow among populations is limited. Finally, the number of generative shoots, the number of seeds per fruit and seed weight were significantly and positively related to population size and local growth conditions. We conclude that under stressful conditions populations of clonal forest plant species can slowly evolve into remnant populations characterized by low levels of genetic variation and limited sexual reproduction. Conservation of suitable habitat conditions is therefore a prerequisite for effective long-term conservation of clonal forest plant species. PMID- 16341716 TI - QSAR for anti-malarial activity of 2-aziridinyl and 2,3-bis(aziridinyl)-1,4 naphthoquinonyl sulfonate and acylate derivatives. AB - Quantitative structure-activity antimalarial relationships have been studied for 63 analogues of 2-aziridinyl and 2,3-bis(aziridinyl)-1,4-naphthoquinonyl sulfonate and acylate derivatives by means of multiple linear regression (MLR) and artificial neural networks (ANN). The antimalarial activity [ log(IC50x10(6))] of the compounds studied were well correlated with descriptors encoding the chemical structure. Using the pertinent descriptors revealed by a stepwise procedure in the multiple linear regression technique, a correlation coefficient of 0.9394 (s=0.2121) for the training set was obtained for the ANN model in a [3-5-1] configuration. The results show that the antimalarial activity of 2-aziridinyl and 2,3-bis(aziridinyl)-1,4-naphthoquinonyl sulfonate and acylate derivatives is strongly dependent on hydrophobic character, hydrogen-bond acceptors and also steric factors of the substituents. PMID- 16341717 TI - Molecular docking study on the "back door" hypothesis for product clearance in acetylcholinesterase. AB - Acetylcholinesterase (AChE) is one of the fastest enzymes known, even though the active site is buried inside the protein at the end of a 20-A deep narrow gorge. Among the great variety of crystal structures of this enzyme, both in the absence and presence of various ligands and proteins, the structure of a complex of AChE with the pseudo-irreversible inhibitor Mf268 is of particular interest, as it assists in the proposal of a back door for product clearance from the active site. Binding of Mf268 to AChE results in the carbamoylation of Ser200 and liberation of an eseroline-fragment as the leaving group. The crystal structure of the AChE-Mf268 complex, however, proves that eseroline has escaped from the enzyme, despite the fact that the Ser-bound inhibitor fragment blocks the gorge entrance. The existence of alternative routes other than through the gorge for product clearance has been postulated but is still controversially discussed in the literature, as an experimental proof for such a back door is still missing. We have used Monte Carlo-based molecular docking methods in order to examine possible alternative pathways that could allow eseroline to be released from the protein after being cleaved from the substrate by Ser200. Based on our results, a short channel at the bottom of the gorge seems to be the most probable back-door site, which begins at amino acid Trp84 and ends at the enzyme surface in a cavity close to amino acid Glu445. [Figure: see text]. PMID- 16341718 TI - Ion pair aggregates and reactions; experiment and theory. AB - Two methods have been developed for determining the aggregation equilibrium constants of lithium enolates based on the change in UV-vis spectrum with concentration and the effect of aggregation on proton-transfer equilibria. Dimers and tetramers are common. Substitution alpha to the carbonyl group generally reduces aggregation. Kinetic studies show that S(N)2 alkylations generally occur with the monomers, even in the presence of large amounts of aggregate. The qualitative chemistry is well modeled by ab initio computations at the HF level with modest basis sets; in these studies solvation is modeled by a combination of coordination of lithium cation with an ether oxygen and the electrostatic interaction of the resulting dipoles and quadrupoles with the solvent dielectric continuum. PMID- 16341719 TI - The ethylene/metal(0) and ethylene/metal(I) redox system: model ab initio calculations. AB - Ab initio calculations (coupled cluster with single and double excitations; CCSD) have been used to investigate the model redox systems ethylene:M(0) (M = Li, Na, K, Rb, Cs) and ethylene:M(I) (M = Be, Mg, Ca, Sr, Ba, Zn, Cd, Hg). Within C2v symmetry, the ground (2A1) states correspond to the charge distribution given in the title. The lowest (2B2) excited states correspond, somewhat counter intuitively, to the ethylene*-/=M(II)ion pair. These trends can be rationalized on the basis of simple electrostatic and configuration-mixing arguments that lead to two simple equations for predicting the electron-transfer energies for oxidation or reduction of the ethylene. The electron-transfer energies to the 2B2 ion pairs are dominated by the electrostatic ion-pairing energies. PMID- 16341721 TI - A possible explanation of the low-level brightness-contrast illusions in the light of an extended classical receptive field model of retinal ganglion cells. AB - The low-level brightness-contrast illusions constitute a special class within visual illusions. Speculations exist that these illusions may be processed through the filtering action of the retinal ganglion cells without necessitating much intervention from higher order processes of visual perception. Concept of the classical receptive field of the ganglion cell, derived from early physiological studies, prompted the idea that a Difference of Gaussian (DoG) model might explain the low-level illusions. In spite of its many successes, the DoG model fails to explain some of these illusions. It has been shown in this paper that it is possible to simulate those illusions with a model that takes into cognizance the role of the extended classical receptive field. PMID- 16341720 TI - [A comparison of the gene expression patterns of human chondrocytes and chondrogen differentiated mesenchymal stem cells for tissue engineering]. AB - BACKGROUND: Tissue engineering is a promising method for the generation of chondrogenic grafts for reconstructive surgery. In cultured chondrocytes, the dedifferentiation of cells seems unavoidable for multiplication. METHODS: In this study, we investigated the expression of distinct markers during the dedifferentiation of human chondrocytes (HC) harvested during septoplasty and human mesenchymal stem cells (hMSC) from cartilage biopsies in cell culture using the microarray technique. RESULTS: The genes for collagen 1alpha1, 2alpha1, 3alpha1, 4alpha1, 11alpha1, biglycan, fibromodulin and lumican were activated during the dedifferentiation of the HCs, collagen 9alpha2, 9alpha3, 10alpha1 and chondroadherin were inactivated. During chondrogenic differentiation of hMSCs, the genes for collagen 3alpha1, 9alpha2, 9alpha3, 10alpha1, 11alpha1 were activated, collagen 4alpha1 and fibromodulin inactivated and the genes for Col 1alpha1, biglycan und chondroadherin constantly expressed. CONCLUSION: The genetic profile for the investigated markers in human chondrocytes generated from hMSCs resembles the profile in differentiated chondrocytes. Collagen 2alpha1, 9alpha2, 9alpha3, 10alpha1 could represent markers for the differentiation of chondrocytes, Col 1alpha1, 3alpha1 und 4alpha1, biglycan, fibromodulin and lumican markers for the dedifferentiation into a more fibroblastoid cell type. PMID- 16341722 TI - Multifractality of decomposed EEG during imaginary and real visual-motor tracking. AB - We test the possible multifractal properties of dominant EEG frequency components, when a subject tracks a path on a map, either only by eyes (imaginary movement - IM) or by visual-motor tracking of discretely moving spot in regular (RM) and Brownian time-step (BM) (real tracking of moving spot). We check the hypotheses that the fractal properties of filtered EEG (1) change with respect to the law of spot movement; (2) differ among filtered EEG components and scalp sites; (3) differ among real and imaginary tracking. Sixteen right-handed subjects begin to perform IM, next--real spot tracking (RM and BM) following a moving spot on streets of a citymap displayed on a computer screen, by push forward/backward a joystick. Multichannel long-lasting EEG is band-pass filtered for theta, alpha, beta and gamma oscillations. The Wavelet-Transform-Modulus Maxima-Method is applied to reveal multifractality [local fractal dimensions Dmax(h)] among task conditions, frequency bands and sites. Non-parametric statistical estimation of the fractal measures h (Dmax) is finally applied. Multifractality is established for all experimental conditions, EEG components and sites as follows among filtered components - anticorrelation (h(Dmax) < 0.5) in beta and gamma, and long-range correlation (h(Dmax) > 0.5) for theta and alpha oscillations; among tasks--for RM and BM, h (Dmax) differ significantly whereas IM resembles mostly RM; among sites--no significant difference for local fractal properties is established. The results suggest that for both imaginary and real visual-motor tracking a line, multifractal scaling, specific for lower and higher EEG oscillations, is a very stable intrinsic one for the activity of large brain areas. The external events (task conditions) insert weak effect on the scaling. PMID- 16341723 TI - Purification and characterization of an enantioselective arylacetonitrilase from Pseudomonas putida. AB - The highly enantioselective arylacetonitrilase of Pseudomonas putida was purified to homogeneity using a combination of (NH4)2SO4 fractionation and different chromatographic techniques. The enzyme has a molecular weight of 412 kDa and consisted of approximately nine to ten identical subunits (43 kDa). The purified enzyme exhibited a pH optimum of 7.0 and temperature optimum of 40 degrees C. The nitrilase was highly susceptible to thiol-specific reagents and metal ions and also required a reducing environment for its activity. These reflected the presence of a catalytically essential thiol group for enzyme activity which is in accordance with the proposed mechanism for nitrilase-catalyzed reaction. The enzyme was highly specific for arylacetonitriles with phenylacetonitrile and its derivatives being the most preferred substrates. Higher specificity constant (kcat/K(m)) values for phenylacetonitrile compared to mandelonitrile also revealed the same. Faster reaction rate achieved with this nitrilase for mandelonitrile hydrolysis was possibly due to the low activation energy required by the protein. Incorporation of low concentration (<5%) of organic solvent increased the enzyme activity by increasing the availability of the substrate. Higher stability of the enzyme at slightly alkaline pH and ambient temperature provides an excellent opportunity to establish a dynamic kinetic resolution process for the production of (R)-(-)-mandelic acid from readily available mandelonitrile. PMID- 16341724 TI - In vitro germination and transient GFP expression of American chestnut (Castanea dentata) pollen. AB - The development of the male reproductive structures of American chestnut (Castanea dentata) is described to advance our understanding of its reproductive behavior. This information has been vital in the development of a strategy to collect pollen grains from male catkins suitable for in vitro germination and transformation experiments. Cutting male catkins into small segments and rolling them over a culture plate resulted in evenly dispersed and large amounts of pollen with minimal unwanted accessory floral parts. To optimize pollen viability, the effect of various storage conditions on in vitro germination was examined. Our results showed that initial storage at 4 degrees C for 2 weeks significantly increased percent germination as compared to freshly collected pollen and those stored directly at -20 degrees C or -80 degrees C. This also means that for long-term storage of American chestnut pollen, the catkins should first be kept at 4 degrees C for a couple of weeks and then at -80 degrees C. The use of pollen grains with high viability is necessary for the transformation of American chestnut pollen. To optimize pollen transformation via particle bombardment, the effects of target distance, target pressure, and pollen developmental stage were examined. Statistical analysis showed that bombardment of ungerminated pollen at 1,100 psi resulted in the highest percent transient GFP expression (4.1%). PMID- 16341726 TI - MicroTom--a high-throughput model transformation system for functional genomics. AB - We have developed a high-throughput Agrobacterium-mediated transformation model system using both nptII and the 5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens strain CP4 (cp4) based selections in MicroTom, a miniature rapid-cycling cherry tomato variety. With the NPTII selection system, transformation frequency calculated as independent transgenic events per inoculated explant ranged from 24 to 80% with an average of 56%, in industrial production scale transformation experiments. For CP4, with glyphosate selection, the average transformation frequency was 57%. Stable transformation frequency was positively correlated with transient expression (R=0.85), and variable with the genes of interest. DNA integration and germline transformation were confirmed by biological assay, Southern Blot analysis, and R(1) phenotype segregation. Transgene expression was observed in leaf, root, stem, flower, and fruit tissues of the transgenic plants. Ninety-five percent of transgenic events coexpressed two introduced genes based on beta-glucuronidase (GUS) and neonmycin phosphotransferase II (NPTII) expression. Seventy-five percent of transgenic events contained one to two copies of the introduced uidA (GUS) gene based on Southern analysis. Transgenic plants from the cotyledon explants to the transgenic plants transferred to soil were produced within about 2-3 months depending on the genes of interest. The utility of this MicroTom model transformation system for functional genomic studies, such as identification of genes related to important agricultural traits and gene function, is discussed. PMID- 16341725 TI - Targeted overexpression of the Escherichia coli MinC protein in higher plants results in abnormal chloroplasts. AB - Higher plant chloroplast division involves some of the same types of proteins that are required in prokaryotic cell division. These include two of the three Min proteins, MinD and MinE, encoded by the min operon in bacteria. Noticeably absent from annotated sequences from higher plants is a MinC homologue. A higher plant functional MinC homologue that would interfere with FtsZ polymerization, has yet to be identified. We sought to determine whether expression of the bacterial MinC in higher plants could affect chloroplast division. The Escherichia coli minC (EcMinC) gene was isolated and inserted behind the Arabidopsis thaliana RbcS transit peptide sequence for chloroplast targeting. This TP-EcMinC gene driven by the CaMV 35S(2) constitutive promoter was then transformed into tobacco (Nicotiana tabacum L.). Abnormally large chloroplasts were observed in the transgenic plants suggesting that overexpression of the E. coli MinC perturbed higher plant chloroplast division. PMID- 16341727 TI - Biophysics of T5, IRA phages, Escherichia coli outer membrane protein FhuA and T5 FhuA interaction. AB - In spite of the similarities in a structural organization of T5 and IRA phages their thermal and hydrodynamical peculiarities are completely different. One of the significant differences is observed in temperature value at which thermally induced DNA ejection starts. If in the case of physiological conditions this difference equals to 30 degrees capital ES, Cyrillic, then it decreases as ionic strength of the solvent decreases. Also, from our experimental results follows that in the opening of phage tail channel for T5 phage (at pH7) significant role play electrostatic forces. In spite of that both of these phages grow on the same Escherichia coli strain, we have shown that these phages need different receptors to penetrate into the bacterial cell precisely FhuA serves as receptor only for T5 phage. The higher FhuA concentration in T5 phage suspension is, the more intensive DNA ejection in environment is. The minimal FhuA/T5 ratio, which is 300/1, correspondingly, necessary for effective DNA ejection from the phage head was experimentally determined. For the first time the ejection of T5 phage DNA induced by FhuA was observed in an incessant regime. The deconvolution of calorimetric curve of FhuA's denaturation has been shown that in a chosen condition there are four thermodynamically independent domains in the structure of FhuA. PMID- 16341728 TI - Prospective study of the long term change of the oral flora after radiation therapy. AB - OBJECTIVES: The aim of this prospective study was to evaluate the long term change in oral pathogens following radiation therapy. METHODS: Twenty-two patients with planned radiation therapy (>30 Gy) of head and neck squamous cell carcinoma were included. Before radiation therapy, after 3, 6, and 12 months samples from the deepest periodontal pocket were drawn. Five major periodontal pathogens were studied using DNA probes (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteriodes forsythus, Treponema denticola). Stimulated saliva was sampled for the microbiologic study of caries pathogens (streptococci and lactobacilli). Plaque index and decayed, missing, filled tooth surfaces (DMF-S) were recorded. RESULTS: A normalisation of the caries bacteria is not found correlating to a significant increase in the number of affected teeth (DMF-S) from 80.7 to 88.5 after 12 months. The plaque index remained unchanged. The incidence of periodontal pathogens did not significantly change during the follow up. CONCLUSION: In contrast to radiation caries there seems to be no microbiological evidence for "radiation periodontitis". Despite of the intensive oral hygiene no reduction of the high number of caries pathogens is found, which leads to a high risk of tooth decay even 12 months after radiation. PMID- 16341729 TI - Antimicrobial activity on glass materials subject to disinfectant xerogel coating. AB - The antimicrobial compound dodecyl-di(aminoethyl)-glycine was immobilized in a silicon oxide xerogel matrix and used for glass surface coating. Coated glasses were tested for surface antimicrobial activity. The utilization of tetraethoxysilane (TEOS) as a silicon oxide polymer precursor, using the dip coating process, allowed for the generation of transparent thin films over glass surfaces. Different concentrations of the antimicrobial compound were used to generate the coatings. The presence of dodecyl-di(aminoethyl)-glycine on coated and uncoated slides was analyzed by FT-IR spectra. Coated glass slides were exposed to suspensions of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus for 24 h. Surface contamination was evaluated by the microbial plate count technique. When antimicrobial-coated glasses were compared with antimicrobial-free coated glasses, the former showed greater than 99% reduction of colony-forming units (cfu) for E. coli and P. aeruginosa, when 1% of antimicrobial was present in the coating solution. The same percentage of reduction for S. aureus was achieved when 1.5% of the antimicrobial was present in the coating solution. In a direct inhibition test on agar plates, no inhibitory zone was observed, indicating that the antimicrobial did not diffuse into the media. PMID- 16341731 TI - Comment on evaluation of neonates with esophageal atresia using chest CT scan. PMID- 16341732 TI - [Fibrocartilaginous embolism of the spinal cord]. PMID- 16341730 TI - [Levobupivacaine for regional anesthesia. A systematic review]. AB - Levobupivacaine [S(-)bupivacaine], the levorotatory S-enantiomer of racemic bupivacaine, is commercially available in the U.S. and in most European countries. We performed a systematic review (MEDLINE database) and identified 88 articles on the clinical application of levobupivacaine in more than 3,000 patients. The use of levobupivacaine is described for epidural, caudal, and spinal anesthesia, for peripheral nerve blocks, for ophthalmic and dental anesthesia, for different pediatric indications and for intravenous regional anesthesia. In these regional techniques, levobupivacaine was used for all common indications in a wide range of clinical settings. Epidural levobupivacaine was combined with fentanyl, morphine, sufentanil, epinephrine, and clonidine, spinal levobupivacaine was combined with sufentanil, fentanyl, and epinephrine. In most studies, levobupivacaine was compared to bupivacaine and/or ropivacaine. PMID- 16341733 TI - [Implementation of a competency-based graduate medical education program in a neurology department]. AB - BACKGROUND: Competency-based programs have long been the basis of graduate medical education in British and North America. Such formally defined program structures are lacking in German graduate medical education. METHODS: The methods and assessment tools used in a curriculum-based program of graduate education in medical specialties are presented. The feasibility of the program was tested in day-to-day use in a pilot study conducted within a neurology department. Experience of it during a 1-year period after implementation has also been collected. RESULTS: It was found that implementation of the program is feasible providing that both the trainers and the students are well motivated, that adequate resources are provided, and that a formal training system is provided for the trainers. This is confirmed by the 1-year experience of the program in routine clinical practice. CONCLUSIONS: The competency-based program presented, which is compatible with many quality management systems, offers management and assessment tools that will make for more satisfactory postgraduate medical education. PMID- 16341735 TI - [Marginal case of a practice fee for psychiatric emergency service]. AB - Since 1 January 2004, the German Act for the Modernisation of Health Insurance (GMG) has regulated collection of a so-called practice fee-a charge to patients for their first outpatient health care in any given quarter. We report the case of a slightly inebriated patient who, against his will, was taken to a psychiatric outpatient department by local authorities, where he had to pay the quarterly charge of 10 EUR. Considering that he was forced and therefore had to pay the fee against his will, the present regulation is being discussed with respect to legal and moral implications in psychiatric emergency cases. PMID- 16341734 TI - [Neurophysiological and neuroimaging studies for restless legs syndrome and periodic leg movement disorder]. AB - The restless legs syndrome (RLS) is a clinical diagnosis based on the four essential criteria defined by the International Restless Legs Syndrome Study Group (IRLSSG). An idiopathic form can be separated from a symptomatic form. Neurophysiological studies have investigated the pathophysiology of the idiopathic RLS or have been used to exclude a symptomatic cause, in particular polyneuropathy. So far cortical excitability changes, corticomotor, somatosensory and auditory pathways, spinal cord excitability, B-wave rhythm and cycling alternating pattern, as well as reflex mechanisms have been investigated by electroencephalography, evoked potentials, Bereitschaftspotentials, nerve conduction and thermal threshold measurements, electromyography, transcranial Doppler sonography, measurements of the spinal flexor reflex as well as neuroimaging techniques. The etiology of the RLS cannot be revealed by these methods, neurophysiological studies in RLS are, however, useful for a better understanding of the pathophysiology and for exclusion of a polyneuropathy or other symptomatic causes. In addition to neurophysiological investigations, small fiber neuropathy, which seems to be a more common finding in RLS patients than expected to date, may need biopsy for confirmation. This review will focus on investigations of the different systems involved with diverse neurophysiological methods. PMID- 16341736 TI - [Plasma exchange therapy for steroid-unresponsive multiple sclerosis relapses: clinical experience with 16 patients]. AB - Patients with severe multiple sclerosis (MS) relapses which do not respond sufficiently to corticosteroids can undergo escalating immunotherapy with plasma exchange. We review the course of 14 apheresis cycles in 13 adult patients and three pediatric cases from our center between 2004 and 2005. Nine cases were due to optic neuritis, five had experienced clinically isolated syndromes, and two suffered from Devic's disease. Of the adult patients, 71% had good or very good outcome. The mean time point of improvement was after the third plasmapheresis session, and early initiation of plasma exchange therapy (within 1 month after begin of relapse) was associated with better outcome. In pediatric MS, two of three patients showed clear improvement. These data argue for a very good therapeutic effect of plasma exchange if performed early and with adequate indication. PMID- 16341737 TI - [Friedrich Christoph Pelizaeus--neurologist and balneology specialist]. PMID- 16341738 TI - The Danish effect on Swedish alcohol costs. An analysis based on hospitalization data from southern Sweden. AB - This study investigated: (a) the cost and change in hospitalizations related to alcohol misuse for the healthcare sector and (b) the effect of distance to the border on alcohol-related hospitalization costs. The first objective was analyzed using descriptive statistics and the second using ordinary least squares regression on aggregated municipality data. The total cost decreased marginally during the study period while the number of patient-cases decreased substantially, presenting evidence of a substitution towards outpatient care. The increase in average treatment cost and the almost constant total cost provide evidence for a societal increase in the burden of alcohol-related diseases. We found a negative effect for distance to Denmark on alcohol-related hospitalization cost for the year 2003. The effect was smaller for 1998, suggesting that the increase in private import quotas during the study period has affected individuals' consumption level and/or consumption pattern. We also found indications that the increase in import quotas lead to a higher cost increase for heavy consumers than for low consumers. PMID- 16341741 TI - Pharmaceutical policy and the pharmacy profession. AB - In this article, the authors look at the relationship between pharmaceutical policy and the pharmacy profession with focus on pharmacy practice and pharmacists in the health care sector. Pharmaceutical policy encompasses three major policy inputs: public health policy, health care policy and industrial policy. In order to analyse and understand pharmaceutical policy, it is important to know how policymakers view pharmacy and pharmacists. The authors look at the issues that arise when policy regulates pharmacy as a business, and what this means for the profession. The perspective of pharmacy as a health care profession, as well as what it means when we view pharmaceutical policy in the context of the health sector labour market, is discussed. The authors also discuss how factors external to the profession are affecting its purpose and realm of practice, including the current trend in managerialism, and how the division of labour with other health professionals such as physicians and pharmacy assistants is affecting the pharmacy profession's position in the labour market. Next the authors look at ways in which the pharmacy profession has affected policy. Pharmacists have been instrumental in developing new and expanding roles for the profession, sometimes inspired by external events, but often as a result of their own prerogative. The pharmacy profession is encouraged to take a leading role in forming and contributing to policy, in this way making visible its contribution to society in general and public health in particular. If not, the profession will forever be reacting to policy and will remain at the mercy of policymakers and other strong actors in society. PMID- 16341742 TI - Adherence of pharmacoeconomic studies to national guidelines in the Netherlands. AB - OBJECTIVE: This study examines the adherence of Dutch pharmacoeconomic studies to the national guidelines of conducting a pharmacoeconomic evaluation. METHODS: Dutch guidelines for pharmacoeconomic research were issued in 1999. All Dutch pharmacoeconomic studies that were published in English during 2000-2002 were selected for our review. Two reviewers examined each study for relevance and compared each study with the nine methodological guidelines selected. RESULTS: It was found that 29 studies satisfied the inclusion criteria. The societal perspective was taken in 13 out of the 29 studies (45%), an adequate time period of analysis was chosen in 21 (72%), effectiveness was explicitly differentiated from efficacy in 17 (59%), an incremental analysis was performed in 23 (79%), costs, benefits and health gains were discounted in 24 (83%), effectiveness was expressed in LYGs or QALYs in 16 (55%), reference prices were used in 8 (28%), subgroup analysis was presented in 13 (45%) and sensitivity analysis was included in 26 (90%). CONCLUSIONS: In this review we found that the adherence of studies to some of the Dutch guidelines for pharmacoeconomic studies is fair. However, major improvements are required with respect to the adoption of the societal perspective, presentation of adequate subgroup analyses and application of reference prices. PMID- 16341743 TI - Using imipenem and cilastatin during continuous renal replacement therapy. AB - AIM: To identify and review studies which have sought to define the pharmacokinetics of imipenem and cilastatin in patients receiving continuous renal replacement therapy (CRRT). METHOD: Literature was primarily identified using Pharmline, Embase and Medline databases using the search terms "imipenem," "haemofiltration," "haemodialysis" and "pharmacokinetics." Papers that discussed only intermittent haemodialysis were excluded. RESULTS: Seven papers were identified which described the pharmacokinetics of imipenem in patients receiving CRRT. Four different modes of CRRT were used. The methods of sampling, dosages used and assumptions made during pharmacokinetic calculations varied widely between the studies. Total body clearance of imipenem during CRRT in patients suffering from acute renal failure was found to range between 89 and 149 ml/min. Total body clearance of cilastatin ranged between 9 and 32 ml/min. Total body clearance of both imipenem and cilastatin was reduced in patients with chronic renal failure. Total body clearance of cilastatin was also reduced by impaired liver function. Dose recommendations made ranged between 500 mg 6-hourly and 500 mg 12-hourly. CONCLUSIONS: The heterogeneity of the studies identified prevents them being analysed as a single group. For meaningful dosage recommendations to be made, further studies are required using larger populations and with more detail regarding liver dysfunction and duration of renal failure. PMID- 16341744 TI - Complex pharmaceutical care intervention in pulmonary care: part A. The process and pharmacists' professional satisfaction. AB - OBJECTIVE: In the IPMP study (Interventions on the principle of Pulmonary Medication Profiles), tailored pharmaceutical care interventions were provided to pulmonary patients selected because of drug use that deviates from Dutch guidelines. The aims were to solve drug-related problems and to improve patients' drug use. This article describes the pharmaceutical care process tailored to the individual problems of patients in the intervention arm of a randomized controlled trial and defines the package of care. METHODS: After a preliminary selection of the patients with the help of the algorithmic IPMP computer instrument, instructed Dutch community pharmacists had structured consultations with patients (aged 13-70 years) in the intervention arm to identify behaviour and specific problems with their medication. Based on this identification process, a tailored intervention was constructed that could comprise one or more of six pharmaceutical care modules. Modules were clustered in sets describing the complete programme of care provided to one patient. If necessary, pharmacists consulted the patients' physicians to improve the prescribed therapy. After the interventions, medication changes were evaluated with the patients. The prescribed medication and the refill rate were monitored in the pharmacy computer during 1 year. All activities and results were extensively monitored and documented. MAIN OUTCOME MEASURE: Process description, i.e. number of provided pharmaceutical care modules and medication changes. Pharmacists' satisfaction. RESULTS: Tailored interventions were provided to 199 patients at risk of sub optimal drug therapy. In all 813 pharmaceutical care modules were performed and documented, and clustered in four different programmes. In addition to education and motivation to adhere to prescribed medication for all 199 patients, a medication change was suggested in 124 cases. Patients and physicians agreed upon a change in 94 cases. Device change was agreed upon in 58 of 64 cases, often simultaneously with medication change. Pharmacists consulted physicians concerning 100 patients. Pharmacists reported satisfaction with the pharmaceutical care approach. CONCLUSION: Because of the extensive documentation, interventions could be described completely. Pharmacists observed a better drug use after educating patients or by solving their drug-related problems. In collaboration with physicians drug treatment could be improved. PMID- 16341745 TI - Complex pharmaceutical care intervention in pulmonary care: part B. Patient opinion and process survey. AB - OBJECTIVE: The IPMP study (Interventions on the principle of Pulmonary Medication Profiles) investigates and describes the results of complex pharmaceutical care interventions provided to selected pulmonary patients to improve their drug use. This paper describes the patients' opinions about the care provided and the results of the intervention. METHOD: Questionnaires investigating patients' opinions about provided pharmaceutical care were sent to 185 patients participating in the IPMP study after the intervention by their pharmacists had been finished. One year after the start of the intervention, patients were invited to a final consultation by their pharmacists to evaluate their drug use and their symptoms (n = 138). At this point in time pharmacists investigated the knowledge of the patients about the medication and their inhaler technique again. MAIN OUTCOME MEASURE: The influence of the intervention on patients' symptoms. Change in drug-related problems, knowledge and skills concerning pulmonary medication after intervention. Satisfaction of the patients with the provided pharmaceutical care. RESULTS: In total 141 out of 185 patients completed the questionnaire. Patients were satisfied with the intervention by their pharmacists and considered it important. The majority (67%) reported that they had learned more about their medication or the inhaler technique, resulting in significantly improved coping behaviour with their pulmonary medication compared with patients who valued the intervention as a nice conversation with their pharmacist only. Patients with improved ability to cope reported statistically significantly fewer symptoms compared with patients reporting no change in behaviour (chi-square test, P < 0.05). In the final consultation of 138 patients, pharmacists observed increased knowledge and skills and decreased drug-related problems. The patients concerned were pleased with the change in treatment and were more satisfied with their current medication as compared with their earlier reports. CONCLUSION: Patients can be influenced effectively by the tailored intervention of pharmacists resulting in improved ability to cope with pulmonary medication and in fewer reported adverse effects and symptoms. Patients attributed these results to the intervention of the pharmacists. PMID- 16341746 TI - The difficulty of opening medicine containers in old age: a population-based study. AB - OBJECTIVES: To investigate elderly people's ability to open medicine containers, and how this ability correlates to some common disorders that may cause functional or cognitive impairment. METHODS: Cross-sectional study of older people age 81 years and older, from the second follow-up (1994-1996) of the Kungsholmen project, a population-based study of very old people in an urban area of Stockholm, Sweden. Six hundred and four persons (mean age 86.7 years) were tested for their ability to open three types of medicine containers. The disorders studied were rheumatoid arthritis, stroke, Parkinson's disease, cognitive impairment (measured by mini-mental state examination, MMSE) and impaired vision. RESULTS: We found that 14% were unable to open a screw cap bottle, 32% a bottle with a snap lid, and 10% a blister pack. Female gender, higher age, living in an institution, Parkinson's disease, rheumatoid arthritis, cognitive impairment and impaired vision were all associated with a decreased ability to open the containers. Less than half of the elderly people who were unable to open one or more of the containers received help with their medication. Among those living in their own homes only 27% received help. CONCLUSION: Older peoples' ability to open medicine containers is impaired by several conditions affecting physical and cognitive functioning. Many elderly people who are unable to open medicine containers do not receive help with their medication, particularly those living in their own homes. PMID- 16341747 TI - When do children convert from liquid antiretroviral to solid formulations? AB - BACKGROUND: Liquid formulations are usually regarded as the gold standard in paediatric formulation; but sometimes, liquid formulations have stability and taste problems as well as being inconvenient for travelling. Therefore, for the management of long-term illness, some older children, parents and clinicians would prefer to use solid formulations. However, there is a lack of studies to investigate the age at which children are converted from liquid to solid formulations. OBJECTIVES: (1) To investigate the age range at which children convert from liquid antiretroviral drug formulations to solid formulations, the formulations are abacavir, didanosine, lamivudine, stavudine, and zidovudine. (2) To calculate how long children stay on each of five UK liquid formulations (retention time) and factors affecting the retention times of the above liquid formulations. METHOD: This was a retrospective medical records survey at Great Ormond Street Hospital for Children, London, United Kingdom. Patients' treatment details were entered into SPSS for Windows v. 11.0 and the retention times for the above liquid formulations were calculated i.e., from initiation of the liquid treatment to conversion to solid preparation. The retention times of different preparations were then compared using Cox regression analysis. RESULTS: A total of 92 patients are included in the analysis. The overall average age at conversion was 7.3 years (95% CI 6.3-8.2). Patients on stavudine were more likely to switch to the corresponding solid dose form than the other four medicines (P < 0.001); more than 50% of patients on stavudine switched to solid formulation after nine months of treatment, however, less than 25% of patients on other formulations switched during the same period. CONCLUSION: Children taking antiretroviral liquid preparations change to solid dose forms at approximately seven years of age. However, for stavudine, children are more likely to take the solid form at an earlier age. PMID- 16341748 TI - The use of lamotrigine and other antiepileptic drugs in paediatric patients at a Malaysian hospital. AB - OBJECTIVE: (1) To determine the effect of lamotrigine add-on therapy on the seizure frequency and cost in paediatric patients. (2) To determine the prescribing pattern of other antiepileptic drugs (AEDs). METHOD: A retrospective study of medical records was carried out from October 2000 to June 2001 at the paediatric clinic, Hospital Pulau Pinang. MAIN OUTCOME MEASURE: Seizure frequency, cost of drug and types of AED prescribed. RESULTS: A total of 209 medical records were retrieved during the study period. Lamotrigine (LTG) was prescribed in 29 patients as add-on therapy. In 18 patients, there was a significant reduction in seizure frequency after the addition of LTG. Approximately 70% experienced a reduction in seizure frequency of more than 50%. Side effects of LTG were considered mild and manageable. However, drug cost after the addition of LTG increased by 103%. In the remaining 180 patients, the most common AED prescribed was sodium valproate (VPA). Only 15% of the patients received combination therapy. Mean monthly cost of monotherapy was found to be RM 24.4 while monthly cost of combination therapy was RM 45.4 (1 Euro-RM 5.00). CONCLUSION: The majority of paediatric patients in the study are on AED monotherapy and only a small percentage was prescribed lamotrigine. The use of lamotrigine is associated with better seizure control but with an increase in drug cost. PMID- 16341749 TI - Development and face validity of explicit indicators of appropriateness of long term prescribing. AB - OBJECTIVES: To develop a set of explicit and operationalisable indicators of appropriate prescribing and assess their face validity using clinical pharmacists practising in secondary and primary care. METHOD: Appropriateness indicators were derived from the literature, applied to data in the hospital clinical records of all newly prescribed long-term drugs for 50 randomly selected patients, further refined and then applied to another 25 randomly selected patients. A pre-piloted postal questionnaire was sent to 200 hospitals and primary care pharmacists, asking them to assess the indicators as to their importance for the assessment of appropriateness of long-term prescribing initiated in hospitals. RESULTS: Fourteen indicators were developed and piloted. Of the 16 original indicators, 5 were discarded, as they were unable to be operationalised, and 2 were subdivided to reflect the routinely available data. Eighty-six pharmacists with individual patient-focussed clinical duties took part in the assessment of the face validity (response rate 43%). Eleven indicators achieved a median importance rating of 1 (very important), and three indicators a median importance rating of 2 on a 5 point scale. The three most important indicators overall were "indication included in discharge summary", "questionable high-risk therapeutic combination" and "hazardous drug-drug combination". CONCLUSION: It was possible to develop and operationalise 14 indicators of the appropriateness of long-term prescribing commenced in hospital practice, all of which were considered to have face validity by an expert panel of clinical pharmacists. The development of these explicit indicators highlighted the incompleteness of the patient's record. Further work is needed to assess their validity and reliability, before their use in research or audit can be recommended. PMID- 16341750 TI - SOFAST-HMQC experiments for recording two-dimensional heteronuclear correlation spectra of proteins within a few seconds. AB - Fast multidimensional NMR with a time resolution of a few seconds provides a new tool for high throughput screening and site-resolved real-time studies of kinetic molecular processes by NMR. Recently we have demonstrated the feasibility to record protein 1H-15N correlation spectra in a few seconds of acquisition time using a new SOFAST-HMQC experiment (Schanda and Brutscher (2005) J. Am. Chem. Soc. 127, 8014). Here, we investigate in detail the performance of SOFAST-HMQC to record 1H-15N and 1H-13C correlation spectra of proteins of different size and at different magnetic field strengths. Compared to standard 1H-15N correlation experiments SOFAST-HMQC provides a significant gain in sensitivity, especially for fast repetition rates. Guidelines are provided on how to set up SOFAST-HMQC experiments for a given protein sample. In addition, an alternative pulse scheme, IPAP-SOFAST-HMQC is presented that allows application on NMR spectrometers equipped with cryogenic probes, and fast measurement of one-bond 1H-13C and 1H 15N scalar and residual dipolar coupling constants. PMID- 16341751 TI - Chemical shifts provide fold populations and register of beta hairpins and beta sheets. AB - A detailed analysis of peptide backbone amide (H(N)) and H alpha chemical shifts reveals a consistent pattern for beta hairpins and three-stranded beta sheets. The H alpha's at non-hydrogen-bonded strand positions are inwardly directed and shifted downfield approximately 1 ppm due largely to an anisotropy contribution from the cross-strand amide function. The secondary structure associated H alpha shift deviations for the H-bonded strand positions are also positive but much smaller (0.1-0.3 ppm) and the turn residues display negative H alpha chemical shift deviations (CSDs). The pattern of (H(N)) shift deviations is an even better indicator of both hairpin formation and register, with the cross-strand H-bonded sites shifted downfield (also by approximately 1 ppm) and with diagnostic values for the first turn residue and the first strand position following the turn. These empirical observations, initially made for [2:2]/[2:4]-type-I' and -II' hairpins, are rationalized and can be extended to the analysis of other turns, hairpin classes ([3:5], [4:4]/[4:6]), and three-stranded peptide beta-sheet models. The H alpha's at non-hydrogen-bonded sites and (H(N))'s in the intervening H-bonded sites provide the largest and most dependable measures of hairpin structuring and can be used for melting studies; however the intrinsic temperature dependence of (H(N)) shifts deviations needs to reflect the extent of solvent sequestration in the folded state. Several observations made in the course of this study provide insights into beta-sheet folding mechanisms: (1) The magnitude of the (H(N)) shifts suggests that the cross-strand H-bonds in peptide hairpins are as short as those in protein beta sheets. (2) Even L-Pro-Gly turns, which are frequently used in unfolded controls for hairpin peptides, can support hairpin populations in aqueous fluoroalcohol media. (3) The good correlation between hairpin population estimates from cross-strand H-bonded (H(N)) shift deviations, H alpha shift deviations, and structuring shifts at the turn locus implies that hairpin folding transitions approximate two-state behavior. PMID- 16341752 TI - Effects of protein-pheromone complexation on correlated chemical shift modulations. AB - Major urinary protein (MUP) is a pheromone-carrying protein of the lipocalin family. Previous studies by isothermal titration calorimetry (ITC) show that the affinity of MUP for the pheromone 2-methoxy-3-isobutylpyrazine (IBMP) is mainly driven by enthalpy, with a small unfavourable entropic contribution. Entropic terms can be attributed in part to changes in internal motions of the protein upon binding. Slow internal motions can lead to correlated or anti-correlated modulations of the isotropic chemical shifts of carbonyl C' and amide N nuclei. Correlated chemical shift modulations (CSM/CSM) in MUP have been determined by measuring differences of the transverse relaxation rates of zero- and double quantum coherences ZQC{C'N} and DQC{C'N}, and by accounting for the effects of correlated fluctuations of dipole-dipole couplings (DD/DD) and chemical shift anisotropies (CSA/CSA). The latter can be predicted from tensor parameters of C' and N nuclei that have been determined in earlier work. The effects of complexation on slow time-scale protein dynamics can be determined by comparing the temperature dependence of the relaxation rates of APO-MUP (i.e., without ligand) and HOLO-MUP (i.e., with IBMP as a ligand). PMID- 16341753 TI - Extended flip-back schemes for sensitivity enhancement in multidimensional HSQC type out-and-back experiments. AB - In many NMR experiments, only polarisation of a limited sub-set of all protons is converted into observable coherence. As recently shown by the "longitudinal" TROSY implementation (Pervushin et al. (2002) J. Am. Chem. Soc., 124, 12898 12902) and SOFAST-HMQC (Schanda and Brutscher (2005) J. Am. Chem. Soc., 127, 8014 8015), recovery of unused polarisation can be used indirectly and unspecifically to cool the proton lattice and, thus, accelerate re-equilibration for the selected proton subset. Here we illustrate transfer of this principle to HSQC based multi-dimensional out-and-back experiments that exploit only polarisation of 15N-bound protons. The presented modifications to the pulse sequences can be implemented broadly and easily, extending standard flip-back of water polarisation to a much larger pool of protons that may comprise all non-15N-bound protons. The underlying orthogonal separation of H(N) polarisation (selected by the main transfer path) from unused H(u) polarisation (flipped-back on the recovery path) is thereby achieved through positive or negative selection by J coupling, or using band-selective pulses. In practice, H(u) polarisation recovery degrades mostly through cumulative pulse imperfections and transverse relaxation; we present, however, strategies to substantially minimise such losses particularly during interim proton decoupling. Depending on the protein's relaxation properties and the extended flip-back scheme employed, we recovered up to 60% H(u) equilibrium polarisation. The concomitant cooling of the proton lattice afforded substantial gains of more than 40%, relative to the water-only flip-back version, in the fast pulsing regime with re-equilibration delays tau much shorter than optimal (tau(opt) = 1.25 x T1(H(N))). These would be typically employed if resolution requirements dominate the total measurement time. Contrarily, if sensitivity is limiting and optimal interscan delays tau(opt) can be set (optimal pulsing regime), the best of the presented flip-back schemes may still afford up to ca. 10% absolute sensitivity enhancement. PMID- 16341755 TI - NMR assignment of HI1506, a novel two-domain protein from Haemophilus influenzae. PMID- 16341754 TI - CASA: an efficient automated assignment of protein mainchain NMR data using an ordered tree search algorithm. AB - Rapid analysis of protein structure, interaction, and dynamics requires fast and automated assignments of 3D protein backbone triple-resonance NMR spectra. We introduce a new depth-first ordered tree search method of automated assignment, CASA, which uses hand-edited peak-pick lists of a flexible number of triple resonance experiments. The computer program was tested on 13 artificially simulated peak lists for proteins up to 723 residues, as well as on the experimental data for four proteins. Under reasonable tolerances, it generated assignments that correspond to the ones reported in the literature within a few minutes of CPU time. The program was also tested on the proteins analyzed by other methods, with both simulated and experimental peaklists, and it could generate good assignments in all relevant cases. The robustness was further tested under various situations. PMID- 16341756 TI - 1H, 13C and 15N backbone and side chain resonance assignments of Haloferax volcanii DHFR1. PMID- 16341757 TI - Resonance assignments of the two N-terminal RNA recognition motifs (RRM) of the human heterogeneous nuclear ribonucleoprotein F (HnRNP F). PMID- 16341758 TI - Backbone resonance assignments for the Fv fragment of catalytic antibody 6D9 complexed with a transition state analogue. PMID- 16341762 TI - A LINE-1 element insertion in the 5'-noncoding region of caprine growth hormone receptor gene. PMID- 16341763 TI - Allozyme variability and phylogenetic relationships in honey bee (Hymenoptera: Apidae: Apis mellifera) populations from Greece and Cyprus. AB - Ten gene enzymic systems (alpha-GPDH, AO, MDH, ADH, LAP, SOD, ALP, ACPH, ME, and EST), corresponding to 12 genetic loci, were assayed from five Greek populations representing three subspecies of Apis mellifera, A. m. cecropia (Pthiotida, Kythira), A. m. macedonica (Macedonia), and the "Aegean race" of A. mellifera, which is supposed to be very similar to A. m. adami (Ikaria, Kasos), as well as a population from Cypus (A. m. cypria). ADH( *)-1, ADH( *)-2, and LAP( *) electrophoretic patterns discriminate the Cyprus population from the Greek populations. MDH( *)-1, EST( *)-3, SOD( *), ALP( *), and ME( *) loci were found to be polymorphic in almost all populations. The observed heterozygosity was found to range from 0.066 to 0.251. Allele frequencies of all loci were used to estimate Nei's genetic distance, which was found to range between 0.011 and 0.413 among the populations studied. UPGMA and neighbor-joining phylogenetic trees obtained by genetic distance matrix methods, as well as a Wagner tree based on the discrete character parsimony method, support the hypothesis that the most distant population is that from Cyprus. Our allozymic data support A. m. cypria as a distinct subspecies, but there was no allozymic support for the distinction of the other subspecies existing in Greece. PMID- 16341764 TI - Analysis of genetic variation in Agerolese cattle breed. PMID- 16341765 TI - Structure and polymorphism analysis of transforming growth factor beta receptor 1 (TGFBR1) in pigs. AB - Many quantitative trait loci (QTL) for growth and reproductive traits have been detected on the porcine chromosome region 1qter (SSC1qter), making it one of the most important genomic regions for pig breeding. SSC1q corresponds to human chromosome 9, on which lies transforming growth factor beta receptor 1 (TGFBR1). We cloned the porcine TGFBR1 cDNA and gene (as a candidate for QTL) and analyzed the gene structure and polymorphism. Porcine TGFBR1 consists of 9 exons and 8 introns. Intron 2 is alternatively spliced at the acceptor site, resulting in two kinds of mRNA, with putative open reading frames of 1500 and 1512 bp in length. The shorter one encodes 499 amino acid residues. The amino acid sequence has 96.2 and 97.2% sequence similarity to those of human and bovine TGFBR1, respectively. The sequence similarity between porcine and murine TGFBR1 is 95.6%. We detected three single-nucleotide substitutions in exons 1, 2, and 7. Those in exons 1 and 7 are nonsynonymous substitutions resulting in Pro8Ser and Ile413Val substitutions, respectively. PMID- 16341766 TI - Leukemia inhibitory factor in follicular fluid is not related to the number and quality of embryos as well as implantation and pregnancy rates. PMID- 16341767 TI - Phylogeny and molecular evolution of Tetraogallus in China. AB - In total, 535 nucleotides in the mtDNA cytochrome b have been sequenced in Tetraogallus. The 53 variable sites define 16 haplotypes in 53 Tetraogallus himalayensis samples, 1 haplotype in 2 T. altaicus samples, and 6 haplotypes in 19 T. tibetanus samples. The lowest genetic distance is between T. himalayensis and T. altaicus (0.011). The divergent time is 0.69 Ma BP between T. himalayensis and T. altaicus, 4.06 Ma BP between T. himalayensis and T. tibetanus, and 4.19 Ma BP between T. altaicus and T. tibetanus. The evidence of this work showed that T. altaicus should fall under the dark-bellied group of Tetraogallus. The dark bellied and white-bellied speciation first occurred along with the uplift of the Qinghai-Tibet Plateau. A maximum glacier (0.80-0.60 Ma BP) led to the formation of T. altaicus. PMID- 16341768 TI - Genetic heterogeneity in Trypanosoma cruzi strains from naturally infected triatomine vectors in northeastern Brazil: epidemiological implications. AB - Eighteen Trypanosoma cruzi strains isolated from naturally infected triatomines were studied genetically. The majority of the strains were from Triatoma brasiliensis, the principal vector of Chagas disease in the northeast of Brazil. Multilocus enzyme electrophoresis (MLEE) and randomly amplified polymorphic DNA (RAPD) analyses were used to investigate the genotypic diversity and the spread of the T. cruzi genotypes in different environments. MLEE clearly distinguished two distinct isoenzyme profiles, and RAPD analysis revealed 10 different genotypes circulating in rural areas. The strains could be typed as isoenzyme variants of the T. cruzi principal zymodeme Z1 (T. cruzi I). An effective program of epidemiological vigilance is required to prevent the spread of T. cruzi I strains into human dwellings. PMID- 16341769 TI - Identification of a single nucleotide polymorphism in porcine testis cytochrome P450-c17 (CYP17) and its effect on steroidogenesis. AB - Raising uncastrated male pigs could have significant economic benefits for pig production. Uncastrated male pigs can accumulate high levels of 16-androstene steroids, however, resulting in boar taint, which is highly objectionable to consumers. Cytochrome P450-c17 (CYP17) interacts with cytochrome b5 in the biosynthesis of the 16-androstene steroids and the sex steroids from pregnenolone. Amino acid substitutions in CYP17 could therefore affect the ability of this enzyme to catalyze the reactions leading to the production of androstenone and the sex steroids. In this study, we established a sensitive and flexible single-stranded conformational polymorphism technique capable of detecting a single nucleotide polymorphism. We then used this method to identify a substitution from T to A at nucleotide 1317 of CYP17, which caused a change in the amino acid sequence from Leu(439) to His(439). This mutation, however, did not alter the enzyme activity of CYP17 in the biosynthesis of androstenone or sex steroids. Other polymorphisms previously suggested for CYP17, which are vital for the functional interaction of CYP17 with CYB5 in human, were not observed. This study suggests that the synthesis of androstenone in pig testis is not directly affected by any polymorphisms in the coding region of the porcine CYP17 gene. PMID- 16341770 TI - Polymorphisms of DNA repair genes: ERCC1 G19007A and ERCC2/XPD C22541A in a northeastern Chinese population. AB - DNA repair systems are responsible for maintaining the integrity of the genome and have a critical role in protecting against mutations that can lead to cancer. DNA repair gene products of ERCC1 and ERCC2/XPD are involved in the nucleotide excision repair pathway. The allele frequencies of the polymorphisms ERCC1 G19007A and ERCC2/XPD C22541A were examined in a northeastern Chinese population. The allele frequencies were 0.21 (A) and 0.79 (G) for ERCC1 G19007A and 0.49 (A) and 0.51 (C) for ERCC2/XPD C22541A. Comparison with average frequencies from previously reported Caucasian studies demonstrated that the A-allele frequency of ERCC1 G19007A was much lower in the northeastern Chinese population, indicating a remarkable ethnic difference (chi((1)) (2) = 160.09, p < 0.001), and that allele frequencies of ERCC2/XPD C22541A showed marginal racial differences (chi((1)) (2) = 4.36, p = 0.04). We have previously reported that both homozygote carriers of the A-allele as well as homozygous carriers of a high-risk haplotype (which includes the AA genotype in ERCC1 G19007A) were at increased risk of basal cell carcinoma, breast cancer, and lung cancer among Caucasians. The low A-allele frequency of ERCC1 G19007A in the Chinese population may suggest that the genetic contribution to cancer risk differs substantially between ethnic groups. PMID- 16341771 TI - Thermolabile variant, PHOX-S, of prophenol oxidase in Drosophila melanogaster. AB - The Phox(S) strain of Drosophila melanogaster is an electrophoretically slow variant found in a wild population at Victoria, Australia. Prophenol oxidase isoform A(1) from PHOX-S was purified and characterized biochemically and genetically. The purified fraction of A(1) from PHOX-S showed a homodimer with a molecular weight of the subunit of approximately 77 kDa. The Phox(S) strain was temperature sensitive in vivo in culture, and the purified protein was thermolabile in vitro. By the deletion mapping method, the Phox(S) locus was cytologically estimated to be at the location 55-A on the right arm of the second chromosome and 79.6 genetically. These data show that PHOX-S is an electrophoretic variant of MOX and that PHOX-S is the first thermolabile protein found in invertebrate prophenol oxidase. PMID- 16341772 TI - Kinetic evidence for the uniport mechanism hypothesis in the mitochondrial tricarboxylate transport system. AB - The kinetics of the transport of citrate by the tricarboxylate transport system located in the inner mitochondrial membrane was studied in proteoliposomes containing the purified carrier protein, in order to verify the previously hypothesized mechanism of uniport (J. Bioenerg. Biomembr. 35, 133-140, 2003) and achieve some information on the kinetic properties of the carrier transport system. For this purpose, a mathematical model has been elaborated and the experimental data were analyzed according to it. The results indicate that the data actually fit with the uniport model, and hence it is confirmed that the carrier has a single binding site for its substrates and can oscillate between the inside and outside form, in both the free and substrate-bound states. The rearrangement of the free form is slower than the bound form in both directions. The dissociation constants for the internal substrate are at least one order of magnitude higher than the one for external citrate. As a consequence of these last two points, the rate of citrate transport by the carrier is much higher when it operates in exchange with another substrate than when it operates in net uniport. PMID- 16341773 TI - ATP synthase of chloroplast thylakoid membranes: a more in depth characterization of its ATPase activity. AB - In contrast to everted mitochondrial inner membrane vesicles and eubacterial plasma membrane vesicles, the ATPase activity of chloroplast ATP synthase in thylakoid membranes is extremely low. Several treatments of thylakoids that unmask ATPase activity are known. Illumination of thylakoids that contain reduced ATP synthase (reduced thylakoids) promotes the hydrolysis of ATP in the dark. Incubation of thylakoids with trypsin can also elicit higher rates of ATPase activity. In this paper the properties of the ATPase activity of the ATP synthase in thylakoids treated with trypsin are compared with those of the ATPase activity in reduced thylakoids. The trypsin-treated membranes have significant ATPase activity in the presence of Ca2+, whereas the Ca2+-ATPase activity of reduced thylakoids is very low. The Mg2+-ATPase activity of the trypsinized thylakoids was only partially inhibited by the uncouplers, at concentrations that fully inhibit the ATPase activity of reduced membranes. Incubation of reduced thylakoids with ADP in Tris buffer prior to assay abolishes Mg2+-ATPase activity. The Mg2+-ATPase activity of trypsin-treated thylakoids was unaffected by incubation with ADP. Trypsin-treated membranes can make ATP at rates that are 75 80% of those of untreated thylakoids. The Mg2+-ATPase activity of trypsin-treated thylakoids is coupled to inward proton translocation and 10 mM sulfite stimulates both proton uptake and ATP hydrolysis. It is concluded that cleavage of the gamma subunit of the ATP synthase by trypsin prevents inhibition of ATPase activity by the epsilon subunit, but only partially overcomes inhibition by Mg2+ and ADP during assay. PMID- 16341774 TI - Two critical factors affecting the release of mitochondrial cytochrome C as revealed by studies using N,N'-dicyclohexylcarbodiimide as an atypical inducer of permeability transition. AB - N,N'-dicyclohexylcarbodiimide (DCCD) was earlier reported to have stimulatory effects on mitochondrial respiration and to induce mitochondrial swelling, when it was added to mitochondrial suspensions. These data seem to imply that DCCD caused the mitochondrial permeability transition (PT), but this possibility had never been investigated. In the present study, effects of DCCD on the mitochondrial structure and function were studied in detail. DCCD was found to induce mitochondrial PT in a cyclosporine A-insensitive manner. Electron microscopic analysis also supported the induction of the mitochondrial PT by DCCD. However, different from many other PT inducers, DCCD failed to cause massive release of mitochondrial cytochrome c. To understand the relationship between the induction of mitochondrial PT and the release of mitochondrial cytochrome c, we compared the actions of DCCD on mitochondrial structure and function with those of Ca2+, known as an ordinary PT inducer. As a result, two parameters considered to be critical for controlling the release of mitochondrial cytochrome c on the induction of PT were mitochondrial volume and the velocity of mitochondrial oxygen consumption. PMID- 16341775 TI - ANT2 isoform required for cancer cell glycolysis. AB - The three adenine nucleotide translocator (ANT1 to ANT3) isoforms, differentially expressed in human cells, play a crucial role in cell bioenergetics by catalyzing ADP and ATP exchange across the mitochondrial inner membrane. In contrast to differentiated tissue cells, transformed cells, and their rho(0) derivatives, i.e. cells deprived of mitochondrial DNA, sustain a high rate of glycolysis. We compared the expression pattern of ANT isoforms in several transformed human cell lines at different stages of the cell cycle. The level of ANT2 expression and glycolytic ATP production in these cell lines were in keeping with their metabolic background and their state of differentiation. The sensitivity of the mitochondrial inner membrane potential (Deltapsi) to several inhibitors of glycolysis and oxidative phosphorylation confirmed this relationship. We propose a new model for ATP uptake in cancer cells implicating the ANT2 isoform, in conjunction with hexokinase II and the beta subunit of mitochondrial ATP synthase, in the Deltapsi maintenance and in the aggressiveness of cancer cells. PMID- 16341776 TI - Identification of a conserved calmodulin-binding motif in the sequence of F0F1 ATPsynthase inhibitor protein. AB - The natural inhibitor proteins IF1 regulate mitochondrial F0F1 ATPsynthase in a wide range of species. We characterized the interaction of CaM with purified bovine IF1, two bovine IF1 synthetic peptides, as well as two homologous proteins from yeast, namely IF1 and STF1. Fluorometric analyses showed that bovine and yeast inhibitors bind CaM with a 1:1 stoichiometry in the pH range between 5 and 8 and that CaM-IF1 interaction is Ca2+-dependent. Bovine and yeast IF1 have intermediate binding affinity for CaM, while the Kd (dissociation constant) of the STF1-CaM interaction is slightly higher. Binding studies of CaM with bovine IF1 synthetic peptides allowed us to identify bovine IF1 sequence 33-42 as the putative CaM-binding region. Sequence alignment revealed that this region contains a hydrophobic motif for CaM binding, highly conserved in both yeast IF1 and STF1 sequences. In addition, the same region in bovine IF1 has an IQ motif for CaM binding, conserved as an IQ-like motif in yeast IF1 but not in STF1. Based on the pH and Ca2+ dependence of IF1 interaction with CaM, we suggest that the complex can be formed outside mitochondria, where CaM could regulate IF1 trafficking or additional IF1 roles not yet clarified. PMID- 16341777 TI - ATP accelerates respiration of mitochondria from rat lung and suppresses their release of hydrogen peroxide. AB - Lung mitochondria were isolated by differential centrifugation from pentobarbital anesthetized male rats. One to three millimolar Mg2+-ATP increased the consumption of oxygen of lung mitochondria oxidizing 10 mM succinate > fourfold (P < 0.01) whereas ATP increased the respiration of liver mitochondria by < 35%. ATP also hyperpolarized partially uncoupled lung mitochondria in the presence of the mitochondria-specific antagonist, oligomycin. However, only 20% of the ATPase activity in the lung mitochondria was blocked by oligomycin compared to a blockade of 91% for liver mitochondria. We investigated the effect of reducing the non-mitochondrial ATPase activity in the lung preparation. A purer suspension of lung mitochondria from a Percoll gradient was inhibited 95% by oligomycin. The volume fraction identified as mitochondria by electron microscopy in this suspension (73.6+/- 3.5%) did not differ from that for liver mitochondria (69.1+/ 4.9%). ATP reduced the mean area of the mitochondrial profiles in this Percoll fraction by 15% (P <0.01) and increased its state 3 respiration with succinate as substrate by 1.5-fold (P < 0.01) with no change in the state 4 respiration measured after carboxyatractyloside. Hence, ATP increased the respiratory control ratio (state 3/state 4, P <0.01). In contrast, state 3 respiration with the complex 1-selective substrates, glutamate and malate, did not change with addition of ATP. The acceleration of respiration by ATP was accompanied by decreased production of H2O2. Thus ATP-dependent processes that increase respiration appear to improve lung mitochondrial function while minimizing the release of reactive oxygen species. PMID- 16341778 TI - Sphingolipids influence the sensitivity of lipid bilayers to fungicide, syringomycin E. AB - Sphingolipids with long chain bases hydroxylated at the C4 position are a requisite for the yeast, Saccharomyces cerevisia, to be sensitive to the ion channel forming antifungal agent, syringomycin E (SRE). A mutant S. cerevisiae strain, Deltasyr2, having sphingolipids with a sphingoid base devoid of C4 hydroxylation, is resistant to SRE. To explore the mechanism of this resistance, we investigated the channel forming activity of SRE in lipid bilayers of varying composition. We found that the addition of sphingolipid-rich fraction from Deltasyr2 to the membrane-forming solution (DOPS/DOPE/ergosterol) resulted in lipid bilayers with lower sensitivity to SRE compared with those containing sphingolipid fraction from wild-type S. cerevisiae. Other conditions being equal, the rate of increase of bilayer conductance was about 40 times slower, and the number of SRE channels was about 40 times less, with membranes containing Deltasyr2 versus wild-type sphingolipids. Deltasyr2 sphingolipids altered neither SRE single channel conductance nor the gating charge but the ability of SRE channels to open synchronously was diminished. The results suggest that the resistance of the Deltasyr2 mutant to SRE may be partly due to the ability of sphingolipids without the C4 hydroxyl group to decrease the channel forming activity of SRE. PMID- 16341780 TI - Annealed silver-island films for applications in metal-enhanced fluorescence: interpretation in terms of radiating plasmons. AB - The effects of thermally annealed silver island films have been studied with regard to their potential applicability in applications of metal-enhanced fluorescence, an emerging tool in nano-biotechnology. Silver island films were thermally annealed between 75 and 250 degrees C for several hours. As a function of both time and annealing temperature, the surface plasmon band at approximately 420 nm both diminished and was blue shifted. These changes in plasmon resonance have been characterized using both absorption measurements, as well as topographically using Atomic Force Microscopy. Subsequently, the net changes in plasmon absorption are interpreted as the silver island films becoming spherical and growing in height, as well as an increased spacing between the particles. Interestingly, when the annealed surfaces are coated with a fluorescein-labeled protein, significant enhancements in fluorescence are observed, scaling with annealing temperature and time. These observations strongly support our recent hypothesis that the extent of metal-enhanced fluorescence is due to the ability of surface plasmons to radiate coupled fluorophore fluorescence. Given that the extinction spectrum of the silvered films is comprised of both an absorption and scattering component, and that these components are proportional to the diameter cubed and to the sixth power, respectively, then larger structures are expected to have a greater scattering contribution to their extinction spectrum and, therefore, more efficiently radiate coupled fluorophore emission. Subsequently, we have been able to correlate our increases in fluorescence emission with an increased particle size, providing strong experiment evidence for our recently reported metal-enhanced fluorescence, facilitated by radiating plasmons hypothesis. PMID- 16341781 TI - The sensitive determination of nucleic acids using fluorescence enhancement of Eu3+-benzoylacetone-cetyltrimethylammonium bromide-nucleic acid system. AB - A new quantitative method for micro amounts of nucleic acids in aqueous solution is proposed using Eu3+-benzoylacetone (BA) complex as fluorescent probe in the presence of cetyltrimethyl-ammonium bromide (CTMAB). Under the optimum condition, the ratio of the fluorescence intensities with and without nucleic acids is proportional to the concentration of nucleic acid in the range of 1.0x10(-9) to 5.0x10(-6) g/mL for herring sperm DNA (hsDNA), 3.0x10(-9) to 1.0x10(-6) g/mL for calf thymus DNA(ctDNA) and 8.0x10(-9) to 1.0x10(-6) g/mL for yeast RNA (yRNA), and their detection limits are 0.33, 0.21 and 0.99 ng/mL, respectively. Actual sample (DNA of Arabidopsis thaliana) was determined satisfactorily. In addition, the interaction mechanism is also investigated. PMID- 16341782 TI - Double labeling and simultaneous detection of B- and T cells using fluorescent nano-crystal (q-dots) in paraffin-embedded tissues. AB - A double immunohistochemical technique for the simultaneous detection of T- and B cells in paraffin-embedded mice tissues have been developed. This procedure is based on using fluorescent nano-crystals (q-dots). The benefit of using q-dots evolves from their unique fluorescence characteristics advantages: such as broad excitation spectrum, narrow emission band and high photo-bleaching threshold compare to organic fluorophores. T cells antigens (CD3) were stained using antibody-coated q-dots with max emission at 655 nm (GalphaRb-QD655). B cells antigens (CD45R/B220) were stained using streptavidin-coated q-dots with max emission at 585 nm (SA-QD585). The simultaneous detection of T- and B cells was demonstrated in paraffin-embedded lymph node using standard fluorescence microscope. PMID- 16341783 TI - Enhancement of europium luminescence in tetracycline-europium complexes in the presence of urea hydrogen peroxide. AB - An increase in the europium emission band was observed, for the first time, with addition of urea hydrogen peroxide to the tetracycline-europium (Tc-Eu)solution. We have observed that the wavelength, the band width and the area of 5D0-->7F2 europium transition change with the urea hydrogen peroxide concentration. We claim that the tetracycline-europium complexes can be used as probes of urea hydrogen peroxide concentration. PMID- 16341784 TI - Studies on interaction of norfloxacin, Cu2+, and DNA by spectral methods. AB - The interactions of norfloxacin (NFA), DNA, and Cu2+ are studied by fluorescence and UV-spectra method. According to the experimental results, it can be concluded that NFA can form a steady binary complex with Cu2+. There is a linear relationship between the Fluorescence intensity of the norfloxacin-Cu2+-DNA system and the concentration of DNA. And when the concentration of the NFA is 1.95x10(-5) mol L-1, they possess a good linearity in the concentration of DNA ranged from 4.7x10(-6) to 2.8x10(-5) mol L-1. It is a good method due to the high sensitivity and selectivity. PMID- 16341785 TI - Fluorimetric determination of histamine in fish using micellar media and fluorescamine as labelling reagent. AB - An analytical method based on the use of fluorescamine to produce a fluorescent derivative with histamine and combined with micellar-enhanced fluorescence detection of the formed complex is developed for the sensitive and rapid determination of histamine in fishes. The fluorescence properties of the obtained complex in water and micellar solutions of sodium dodecyl sulfate (SDS), cetyltrimethylammonium chloride (CTAC) and brij-700 are reported. Physicochemical variables influencing the sensitivity of the method (pH, micellar, fluorescamine and NaCl relative concentrations) have been optimized. The stability of the formed complex, as shown by kinetic study, depends on the pH of the solution. Linear calibration curves allowing an effective histamine determination were established with large linear dynamic range (LDR), and low limits of detection (LOD) between 0.5 and 33 ng mL-1, according to the solvent. Application to the analysis of fish samples (sardines) yielded satisfactory results. The method seems to be suitable for environmental fish quality control. PMID- 16341786 TI - Ground and excited states proton transfer reactions of 1,8-diaminonaphthalene in perchloric acid solutions. AB - The proton-transfer reaction of 1,8-diaminonaphthalene (1,8-DAN) in acidic medium was studied by means of fluorescence and picosecond spectroscopic techniques. It has been found that there are three different forms of 1,8-DAN in the ground state, but only two different forms in the excited state. The absorption of the mono-cation form of 1,8-DAN is found to be a mixture of the neutral form and the di-cation form. However, the emission is found to be the same as the neutral form, due to the fast dissociation of the mono-cation form once it is excited. The fluorescence of the mono-cation form of 1,8-DAN shows a small shift under different excitation wavelengths. The di-cation form only fluoresces if no free water cluster is available as a proton acceptor. The reaction in the excited state is shown to be a diabatic quenching reaction. With the help of quantum yields and fluorescence lifetime measurements these results are interpreted in terms of a new photochemical scheme. All dissociation and quenching rate constants, pKa and kq, have been determined. PMID- 16341787 TI - Laser-induced fluorescence imaging and spectroscopy of GFP transgenic plants. AB - Green fluorescent protein (GFP) and other fluorescent protein bioreporters can be used to monitor transgenes in plants. GFP is a valuable marker for transgene presence and expression, but remote sensing instrumentation for stand-off detection has lagged behind fluorescent protein marker biotechnology. However, both biology and photonics are needed for the monitoring technology to be fully realized. In this paper, we describe laser-induced fluorescence imaging and laser induced fluorescence spectroscopy of GFP-transgenic plants in ambient light towards the application of remote sensing of transgenic plants producing GFP. PMID- 16341788 TI - Improved fluorescent proteins for single-molecule research in molecular tracking and co-localization. AB - Three promising variants of autofluorescent proteins have been analyzed photophysically for their proposed use in single-molecule microscopy studies in living cells to compare their superiority to other fluorescent proteins previously reported regarding the number of photons emitted. The first variant under investigation the F46L mutant of eYFP has a 10% greater photon emission rate and >50% slower photobleaching rate on average than the standard eYFP fluorophore. The monomeric red fluorescent protein (mRFP) has a fivefold lower photon emission rate, likely due to the monomeric content, and also a tenfold faster photobleaching rate than the DsRed fluorescent protein. In contrast, the previously reported eqfp611 has a 50% lower emission rate yet photobleaches more than a factor 2 slowly. We conclude that the F46L YFP and the eqfp611 are superior new options for single molecule imaging and tracking studies in living cells. Studies were also performed on the effects of forced quenching of multiple fluorescent proteins in sub-micrometer regions that would show the effects of dimerization at low concentration levels of fluorescent proteins and also indicate corrections to stoichiometry patterns with fluorescent proteins previously in print. We also introduce properties at the single molecule level of new FRET pairs with combinations of fluorescent proteins and artificial fluorophores. PMID- 16341789 TI - A new fluorescent assay for enalapril maleate. AB - A new spectrofluorimetric method for the enalapril maleate monitoring was studied. Enalapril maleate was found to be highly photolabile. This drug was evaluated according to photodegradation assay at pH 2.5 and 6. Enalapril maleate was exposed to UVA-UVB radiations. Under these specific conditions was found as degradation product, the diketopiperazine. The modification of the fluorescent properties of enalapril maleate in solution after exposure UV-radiation and the degradation mechanisms were studied. The photodegradation was followed by the developed spectrofluorimetric assay. PMID- 16341790 TI - The fluorescence bioassay platforms on quantum dots nanoparticles. AB - In this paper, we present the optical properties and the platforms on fluorescent quantum dots for biological labeling, biomedical engineering and biosensor in molecular imaging. Quantum dots possess several properties that make them very attractive for fluorescent tagging: broad excitation spectrum, narrow emission spectrum, precise tunability of their emission peak, longer fluorescence lifetime than organic fluorophores and negligible photobleaching. We describe how to take such advantages of quantum dots to develop the technology and employ it to build assay platforms. Finally, ultrasensitivity, multicolor, and multiplexing of the technology of semiconductor quantum dots open up promising and interesting possibilities for bioassay platform. PMID- 16341791 TI - Determination of catechin in aqueous solution by chemiluminescence method. AB - A method to determine catechin in aqueous solution by measuring chemiluminescence intensities using a stopped flow system has been studied. The lucigenin-hydrogen peroxide chemiluminescence reaction was chosen for the determination of catechin. Fe(II) ion was added to the chemiluminescence system to increase the sensitivity. The chemiluminescence intensity from the lucigenin system was increased by the addition of catechin. Effects of flow rates of reagent and sample and concentrations of lucigenin, hydrogen peroxide, Fe(II) ion and KOH were investigated. The calibration curve for catechin was linear over the range from 1.0x10(-6) to 1.0x10(-3) M and the detection limit was 3.0x10(-7) M under the optimal experimental conditions. PMID- 16341792 TI - Fluorescence dynamics of DNA condensed by the molecular crowding agent poly(ethylene glycol). AB - Condensation of extended DNA into compact structures is encountered in a variety of situations, both natural and artificial. While condensation of DNA has been routinely carried out by the use of multivalent cations, cationic lipids, detergents, and polyvalent cationic polymers, the use of molecular crowding agents in condensing DNA is rather striking. In this work, we have studied the dynamics of plasmid DNA condensed in the presence of a molecular crowding agent, polyethylene glycol (PEG). Steady-state and time-resolved fluorescence of the recently established condensation-indicating DNA binder, YOYO-1 [G. Krishnamoorthy, G. Duportail, and Y. Mely (2002), Biochemistry 41, 15277-15287] was used in inferring the dynamic aspects of DNA condensates. It is shown that DNA condensed by PEG is more flexible and less compact when compared to DNA condensed by binding agents such as polyethyleneimine. The relevance of such differences in dynamics toward functional aspects of condensed DNA is discussed. PMID- 16341793 TI - Fluorescent aggregates of 1-(p-butyloxyphenyl)-4-(p-cyanophenyl)buta-1E,3E-diene: temperature sensing and photoimaging applications. AB - The use of fluorescent solid films of a 1-(p-butyloxyphenyl)-4-(p cyanophenyl)buta-1E, 3E-diene as an efficient temperature sensor is described. The fluorescence changes in these films are proposed to occur due to a reversible temperature-dependent variation in its monomer-aggregate ratio. Photoisomerization of the butadiene chromophore in solid films was observed to occur only at elevated temperatures (>120 degrees C), making the material also useful for reversible photochemical generation of fluorescence patterns with nondestructive readout properties. PMID- 16341794 TI - Nonenzymatic direct assay of hydrogen peroxide at neutral pH using the Eu3Tc fluorescent probe. AB - A detailed study is presented on the use of an easily accessible probe (the europium-tetracycline 3:1 complex; referred to as Eu3Tc) for determination of hydrogen peroxide (HP). Eu3Tc undergoes a 15-fold increase in luminescence intensity on exposure to an excess of HP. Data are given on the time dependence of the reaction, on the pH dependence of the absorption and emission spectra of both the probe and its complex with HP, and on the effect of stoichiometry between Eu3+ and Tc on selectivity and signal change. HP can be quantified in aqueous solution of pH 6.9 over a 2-400 microM concentration range with a limit of detection of 960 nM. The assay is validated using standard additions, and mean recoveries are found to be between 97.0 and 101.8%. Species that interfere in concentrations below 1 mM include phosphate, copper(II), fluoride and citrate. The addition of detergents causes the response curves towards HP to shift to higher HP concentrations. The method is critically assessed with respect to other common optical methods for determination of HP. PMID- 16341796 TI - The Anthracen-9-ylmethyloxy unit: an underperforming motif within the fluorescent PET (photoinduced electron transfer) sensing framework. AB - Compound, which was designed to act as a fluorescent sensor for calcium according to the PET (Photoinduced Electron Transfer) principle, shows a relatively small Ca2+-induced fluorescence enhancement factor (FE) of 1.8 whereas its close relative is known to display a far higher FE value of 16. Though designed as fluorescent PET sensors for solvent polarity, compounds and also show negligible fluorescence enhancement as their environments are made progressively less polar even though their relatives and show limiting FE values of 53 and 3, respectively. Indeed, and are useful since they are fluorescent sensors for solvent polarity without being affected by Bronsted acidity. The poor sensory performance of and relative to their cousins is attributed to the presence of an oxygen proximal to the 9-position of an anthracene unit, which opens up a CT (charge transfer) channel. Normal PET sensing service is resumed when the offending oxygen is deleted. PMID- 16341795 TI - Electroporative adjustment of pH in living yeast cells: ratiometric fluorescence pH imaging. AB - A number of vital cell functions including modulation of signaling pathways and regulation of the cellular transport critically depends on the cytoplasmic pH. Many pathological cellular changes are related to the abnormal cytosolic pH as well. Reliable and well-calibrated methods for quantification of the cytosolic pH are therefore of high importance. The pH calibration is particularly difficult in walled cells since standard methods fail. In this report we evaluated the new electroporative calibration method of the cytosolic pH in yeasts by the fluorescence microscopy. The calibration was done on living cells using pyranine as a ratiometric pH-sensitive probe. The probe was electroporatively delivered to the cytosol. We have shown that unlike the measurements in suspension the fluorescence microscopy reveals cell subpopulations with different sensitivity to the pH calibration. While the majority of the cells were well calibrated, there was found subpopulation of uncalibrated cell as well as singular cells exhibiting anomalous pH calibration due to the staining of acidic organelles. Resolution of cell subpopulations helps to achieve better pH calibration compared to the calibration in cuvette on a cell suspension. PMID- 16341797 TI - Enhanced fluorescence from periodic arrays of silver nanoparticles. AB - Electron beam lithography was used to fabricate silver nanoparticle arrays and study the effects of geometrical properties of particles on metal-enhanced fluorescence. Nanoparticle size, shape, interparticle spacing, and nominal thickness were varied in a combinatorial pattern for investigation of the particle plasmon resonance effect on enhancement of fluorescence from three different fluorophores; Fluorescein, Cy3, and Cy5. A specific geometric property for optimal enhancement from each fluorophore was determined. For interparticle spacings greater or equal to 270 nm, the enhancement matched what is expected for a single-particle model. For those particles smaller than 210 nm, the enhancement was lower than for the larger spacing in the range studied. Triangular-shaped particles gave similar enhancement to those of square-shaped particles. This combinatorial pattern by e-beam lithography was found to be useful for studying how individual parameters enhance the fluorescence that are important for rational design of enhanced fluorescence sensors. PMID- 16341799 TI - On the quantitative treatment of donor-donor energy migration in regularly aggregated proteins. AB - An algorithm is presented that quantitatively accounts for donor-donor energy migration (DDEM) among fluorophore-labeled proteins forming regular aggregates. The DDEM algorithm is based on Monte Carlo and Brownian dynamics simulations and applies to calculation of fluorescence depolarisation data, such as the fluorescence anisotropy. Thereby local orientations, as well as reorienting motions of the fluorescent group are considered in the absence and presence of DDEM and among, in principle, infinitely many proteins as they form regular aggregates. Here we apply the algorithm for calculating and illustrating the DDEM and the time-resolved fluorescence anisotropy under static as well as dynamic conditions within helical, linear and circular aggregate structures. A principal approach of the DDEM algorithm for analysing protein aggregates is also outlined. PMID- 16341798 TI - Membrane organization and dynamics of the G-protein-coupled serotonin1A receptor monitored using fluorescence-based approaches. AB - The G-protein-coupled receptor (GPCR) superfamily represents one of the largest classes of molecules involved in signal transduction across the plasma membrane. Fluorescence-based approaches have provided valuable insights into GPCR functions such as receptor-receptor and receptor-ligand interactions, real-time assessment of signal transduction, receptor dynamics on the plasma membrane, and intracellular trafficking of receptors. This has largely been possible with the use of fluorescent probes such as the green fluorescent protein (GFP) from the jellyfish Aequoria victoria and its variants. We discuss the potential of fluorescence-based approaches in providing novel information on the membrane organization and dynamics of the G-protein-coupled serotonin1A receptor tagged to the enhanced yellow fluorescent protein (EYFP). PMID- 16341800 TI - Polar plot representation for frequency-domain analysis of fluorescence lifetimes. AB - We present applications of polar plots for analyzing fluorescence lifetime data acquired in the frequency domain. This graphical, analytical method is especially useful for rapid FLIM measurements. The usual method for sorting out and determining the underlying lifetime components from a complex fluorescence signal is to carry out the measurement at multiple frequencies. When it is not possible to measure at more than one frequency, such as rapid lifetime imaging, specific features of the polar plot analysis yield valuable information, and provide a diagnostic visualization of the participating fluorescent species underlying a complex lifetime distributions. Data are presented where this polar plot presentation is useful to derive valuable, unique information about the underlying component distributions. We also discuss artifacts of photolysis and how this method can also be applied to samples where each fluorescence species shows a continuous distribution of lifetimes. Polar plots of frequency-domain data are commonly used for analysis of dielectric relaxation experiments (Cole Cole plots), which have proved to be exceptionally useful in that field for decades. We compare this analytical tool that is well developed and extensively used in dielectric relaxation and chemical kinetics to fluorescence measurements. PMID- 16341802 TI - BAP1 and breast cancer risk. AB - BAP1 whose protein interacts with BRCA1 was analysed in a series of 47 French familial breast cancer cases negatively tested for BRCA1/2 mutations. The lack of detection of deleterious mutations suggests that BAP1 is not a high risk breast cancer predisposing gene. However, a common identified variant, rs123602, may be tested in sporadic cases as candidate for moderate risk. PMID- 16341803 TI - Patient satisfaction of BRCA1/2 genetic testing by women at high risk for breast cancer participating in a prevention trial. AB - With the increasing availability of cancer risk counseling and genetic testing, we need to determine the most effective way to provide complex and sensitive information to patients. This study was designed to determine the satisfaction of results delivery in women who participated in a breast cancer prevention trial and chose to undergo free and confidential BRCA1/2 genetic testing. Self-selected women at high-risk for breast cancer who were eligible to participate in a phase II chemoprevention trial, were offered free and confidential pre-test counseling and BRCA1/2 full sequencing. Subjects were not randomized but rather had the option of in person or telephone results disclosure. Those subjects with an identified germline alteration were required to follow-up with an in person consultation; this was optional for those with a negative result. A satisfaction survey was mailed to subjects after receiving their results. Ninety-seven percent (116/119) of the eligible subjects underwent genetic testing. Ninety-one percent (105/116) of those women tested responded to the follow-up survey. Twenty-four of the 26 women with an identified germline alteration responded. Nearly all of the responders were satisfied with the counseling and testing process. All of the respondents felt they made a wise decision in having the testing and would recommend that other women in a similar situation undergo genetic testing. We found that the majority of women at high risk for breast cancer participating in a prevention trial will choose to undergo anonymous and free BRCA1/2 genetic testing, be informed of the results, and are accepting of receiving results initially by phone. PMID- 16341805 TI - Mutational analysis of hMsh6 in Israeli HNPCC and HNPCC-like families. AB - Germline mutations in DNA mismatch repair (DNA-MMR) genes, mainly hMlh1 and hMsh2, underlie Hereditary Non-Polyposis Colorectal Cancer (HNPCC). Germline hMSH6 gene mutations have been reported in a small subset of HNPCC families. In the present study, ethnically diverse individuals with HNPCC and HNPCC-like features were genotyped for hMsh6 germline mutations using exon-specific PCR, DGGE, and DNA sequencing. The study encompassed 92 individuals representing 88 unrelated families who were previously analyzed for Msh2 and Mlh1 mutations: Jewish Ashkenazim (n = 44), non-Ashkenazim (n = 27), Israeli Moslem-Arab (n = 15), Druze (n=3), and Cypriot non-Jews (n = 3). Of the study population, 71 had colon cancer (CRC), mean age at diagnosis was 50.9+/-13.2 years (range 16-73 years), 5 had endometrial cancer (two with concurrent CRC), (mean 43.6+/-3.26 years, range 38-45 years), and unaffected individuals (n = 18) were first degree relatives within HNPCC families and were genotyped at a mean age of 48.3+/-11.7 years (range 30-69 years). Of the 92 individuals analyzed, none showed a truncating hMsh6 mutation, and 6 (6.6%) harbored one of three germline missense mutations: a previously reported one (V878A), and two novel mutations (V509A, S227I). The pathogenic significance of these three missense mutations is yet unclear. In addition, 5 polymorphisms were detected, 2 of which were novel. We conclude that the rate of pathogenic hMsh6 mutations in HNPCC families of Jewish and Mediterranean origin is low, and that mutations in other genes probably account for the phenotype in these families. PMID- 16341804 TI - MLH1 and MSH2 mutations in Colombian families with hereditary nonpolyposis colorectal cancer (Lynch syndrome)--description of four novel mutations. AB - This study searched for mutations in the MLH1 and MSH2 genes in 23 unrelated Colombian families with suspected hereditary nonpolyposis colorectal cancer (HNPCC). The families were grouped according to the fulfillment of the Amsterdam II criteria or the Bethesda guidelines. We screened all probands by single-strand conformational polymorphism (SSCP) and direct DNA sequencing. Eleven families fulfilled the Amsterdam criteria II and 12 families the Bethesda guidelines. Germline mutations were detected in 11 families, which corresponds to a mutation detection rate of 48%. When only families fulfilling the Amsterdam II criteria were analyzed, the mutation detection rate rose to 82%. Only 8% of the mutation detection rate was found in families following the Bethesda guidelines. Three mutations were shared by two different families, which corresponds to a total of eight different mutations, seven of them found in the MLH1 gene and one in the MSH2 gene. We have identified four mutations that have not been previously reported to the International Collaborative Group of HNPCC. Three of these are pathogenic, a single base substitution (C > T) at codon 640, exon 17, a G deletion at codon 619, exon 16 and in the MLH1 gene and a two-nucleotide deletion (TG) at codon 184, exon 3 in the MSH2. Also, an unclassified variant, a substitution (C > G) at the codon 141, exon 5 of the MLH1, was detected. PMID- 16341806 TI - Long term follow-up of HNPCC gene mutation carriers: compliance with screening and satisfaction with counseling and screening procedures. AB - Hereditary non polyposis colorectal cancer (HNPCC) is a hereditary predisposition to colorectal and endometrial cancer, caused by mutations of the mismatch repair (MMR) genes MSH2, MLH1 and MSH6. Regular colonoscopy reduces the incidence of colorectal cancer in mutation carriers dramatically. The aim of this study was to evaluate the use of colonoscopy by proven HNPCC mutation carriers. We also evaluated the satisfaction with the counseling and screening procedures at the long term. A questionnaire survey was performed among 94 proven MMR gene mutation carriers. Data were analyzed using univariate and multivariate analysis. The average time of follow-up was 3,5 years (range 0.5-8.5 years). The response rate was 74%. The proportion of unaffected mutation carriers under colonoscopic screening increased from 31 to 88% upon genetic testing, and for gynecological screening from 17 to 69%. However, more than half of the responders experienced colonoscopy as unpleasant or painful. About 97% felt well informed during counseling, and 88% felt sufficiently supported. Ten percent of the responders reported a high cancer worry that was significantly (P = 0.007) associated with a high perceived cancer risk. Six responders (9%) regretted being tested. Remarkably, of 4 of these 6 a close relative died recently of cancer. Problems with obtaining a disability or life insurance or mortgage were experienced by 4 out 10 healthy carriers opting for these services. In conclusion, genetic testing for HNPCC considerably improves compliance for screening, which will result in a reduction of HNPCC-related cancer morbidity and mortality in mutation carriers. Most HNPCC gene mutation carriers cope well with their cancer susceptibility on the long term. PMID- 16341807 TI - Survival of patients with ovarian cancer due to a mismatch repair defect. AB - PURPOSE: Hereditary non-polyposis colorectal cancer (HNPCC, Lynch syndrome) is characterized by the development of cancer of the colorectum, endometrium and other cancers. Cancer of the ovaries (OC) has frequently been reported in HNPCC. Colorectal cancer associated with HNPCC has a better survival chance compared to sporadic colorectal cancer. It is yet unknown whether patients with OC from HNPCC families (OC-HNPCC) also have a better survival. Therefore, the aim of the study was to compare the survival between patients with OC-HNPCC and a control group. METHODS: A total of 26 patients with OC were identified from the Dutch HNPCC Registry. A control group (52 cases) matched for age, stage and year of diagnosis was derived from the population-based Eindhoven Cancer Registry. Data on treatment were collected for all patients. Kaplan-Meier analysis was used to calculate the crude survival. RESULTS: The mean age at diagnosis of OC-HNPCC was significantly lower than the age of sporadic OC (49.5 vs 60.9 years). Compared to sporadic OC, OC-HNPCC was diagnosed at an earlier stage. The survival rate was not significantly different between patients with OC-HNPCC and the controls with sporadic OC. The cumulative 5-year-survival rates were 64.2 and 58.1% respectively. CONCLUSION: On the basis of our findings, we recommend to treat OC HNPCC similar to sporadic OC. PMID- 16341808 TI - Behavioral and economic impact of a familial history of cancers. AB - BACKGROUND: Misunderstanding of cancer screening recommendations or messages and confidence in the predictive value of positive familial history of disease may converge to stimulate an over-utilization of screening tests in oncology by patients who perceive themselves to be at high risk. METHODS: A survey looking for predictors of the uptake of five cancer screening tests (mammography, colonoscopy, Fecal Occult Blood Test, upper digestive tract endoscopy and chest X ray) was carried out on 4000 healthy adults (mean age 46.4 years). FINDINGS: Based on the results of a multivariate analysis, the survey enlightens the existing relationships between familial history and increasing uptake of medical cancer screening tests, with OR ranging from 1.3 (IC 1.0-1.6) for chest X-ray to 3.0 (IC 2.1-4.1) for colonoscopy. In France (60 million inhabitants), a conservative assessment of the annual net number of unhelpful screening tests attributable to positive family history of related cancer with chest X-ray and Upper digestive tract endoscopy lead to a figure of 7000 and 7800 tests respectively corresponding to a total annual cost of more than Euro 7, million. INTERPRETATION: Clearer messages about hereditary risks and transparency about the efficacy of screening tests are required in order to decrease over utilization of screening tests and their related costs. PMID- 16341809 TI - Linkage of a pedigree drawing program and database to a program for determining BRCA mutation carrier probability. AB - An export module was developed for transferring primary relational pedigree data directly from a widely used pedigree program and relational database to the BRCAPRO program for calculation of BRCA gene mutation probability estimation. PMID- 16341810 TI - Lack of germ-line mutations at the specific BRCA1-IRIS coding sequence in 114 Spanish high-risk breast/ovarian families. AB - A new BRCA1 locus product called BRCA1-IRIS has been identified recently. High risk breast/ovarian families have not been screened for germ-line mutations at the specific BRCA1-IRIS coding sequence, as it was considered merely as part of BRCA1 intron 11. Here we report the first comprehensive screening of germ-line mutations in a cohort of 116 index cases from high-risk breast/ovarian families in which no germ-line mutation was identified in BRCA1 or BRCA2. We did not find germ-line mutations at the specific BRCA1-IRIS coding sequence in any sample. The only heterozygous patter identified by DGGE was caused by a C to A substitution in the non-coding 3' sequence, 123 bases downstream of the BRCA1-IRIS stop codon (IVS11+268C/A). The data indicates that it is probably a neutral change not associated with cancer risk. Our analysis suggests that the role of germ-line mutations at the specific BRCA1-IRIS sequence in breast cancer susceptibility, if any, is marginal and do not explain a significant fraction of high-risk breast/ovarian families, at least in the population analyzed. PMID- 16341811 TI - Neurofibromatosis 2 (NF2) and malignant mesothelioma in a man with a constitutional NF2 missense mutation. AB - Neurofibromatosis 2 (NF2) is caused by inactivating mutations of the NF2 tumor suppressor gene. Somatic NF2 mutations also occur in a high proportion of human primary malignant mesotheliomas. We report an elderly man with NF2, malignant mesothelioma, and a constitutional NF2 missense mutation. The long latent period for mesothelioma in this patient (61 years) raises the possibility that the type of mutant NF2 allele could affect mesothelioma tumorigenesis or progression. PMID- 16341812 TI - Syndrome of early onset colon cancers, hematologic malignancies & features of neurofibromatosis in HNPCC families with homozygous mismatch repair gene mutations. AB - Hereditary nonpolyposis colon cancer (HNPCC) is the most common hereditary colon cancer syndrome. It is characterized by multiple colon as well as extracolonic cancers such as endometrial, ovarian and urinary tract cancers. In addition, it is well known that some cases of HNPCC can present with unique tumor spectrums such as sebaceous tumors, which is often referred to as the 'Muir-Torre' syndrome. In recent years there have been a few reports of families presenting with early onset of colon tumors along with cafe-au-lait spots and/or hematologic malignancies often associated with homozygous mutations involving one of the mismatch repair genes. In this article we have performed a comprehensive review of the entire medical literature to identify all cases with similar presentations reported in the literature and have summarized the clinical features and genetic test results of the same. The available data clearly highlight such presentations as a distinct clinical entity characterized by early onset of gastrointestinal tumors, hematologic malignancies as well as features of neurofibromatosis (easily remembered by the acronym ;CoLoN'; Colon tumors or/and Leukemia/Lymphoma or/and Neurofibromatosis features). Furthermore, there has also been some evidence that the neurofibromatosis type-1 gene is a mutational target of the mismatch repair deficiency that is seen in families with HNPCC, and that mlh1 deficiency can accelerate the development of leukemia in neurofibromatosis (Nf1) heterozygous mice. Recognition of this syndrome has significant importance in terms of earlier detection of cancers, cancer screening recommendations as well as genetic counseling offered to such families. PMID- 16341816 TI - The meandering mesenteric artery. PMID- 16341817 TI - [Secondarily accelerated foreign bodies as a source of danger from airbag deployment]. AB - BACKGROUND: The use of airbag systems in motor vehicle construction has led to a significant reduction in both the frequency and intensity of injuries during traffic accidents. However, recent reports have shown numerous cases in which it is assumed that the airbag was the major cause of serious injury. CASE REPORT: In a collision with a wild boar, both airbags in the automobile deployed. The driver suffered facial abrasion and belt contusion to the left shoulder. The passenger held a glass bottle in her hands during the impact that, secondarily accelerated through the airbag, hit both eyes and the bridge of her nose. On clinical admission, the patient showed fractures of the nasal bridge, of both medial orbital walls on both sides and a massive periocular haematoma which required direct examination under narcosis. Both eyes showed an orbital contusion, a large area of corneal abrasion, traumatic mydriasis with sphincter tears such as bilateral 90% hyphema. During surveillance, a progressive reduction in visual acuity occurred. After posterior chamber vitrectomy with epiretinal peeling and posterior chamber lens implantation in the left eye, visual acuity was 0.05 in the left eye and 0.01 in the right. CONCLUSIONS: A considerable danger emanates from objects which are situated between head and airbag in the moment of airbag deployment. This requires public education and, in addition, airbag systems which are adapted to the drivers height and weight, as well as to objects in its immediate vicinity. PMID- 16341818 TI - [Objectivity of therapeutic results following skull base surgery using virtual model analysis]. AB - BACKGROUND: Virtual model analysis of patient head tracking allows for objectivity and the monitoring of therapeutic results of pathologies in the skull base region. The introduction of these models in clinical routine has been impaired by the extended time needed for the preparation of radiological data. METHODS: Quality control analysis was carried out for seven cases with different pathological findings in the skull base region in patients who had undergone virtual model analysis. RESULTS: Preparation time of radiological data for the process of segmentation required, under optimal conditions, a minimum of 30 min. Virtual model analysis enables spatial visualization of regions of interest and adjacent anatomical structures. This improves case-specific pathoanatomical understanding as well as preoperative planning of surgical strategies. CONCLUSIONS: Virtual model analysis improves the physician's spatial comprehension of localized pathological findings at the dysmorphic interface of bone and soft tissue across the skull base. Therefore, it seems to be an adequate tool for quality control analysis of therapeutic results after extended skull base surgery. PMID- 16341819 TI - Molecular evidence demonstrating the basidiomycetous fungus Cryptococcus curvatus is the dominant microbial eukaryote in sediment at the Kuroshima Knoll methane seep. AB - The Kuroshima Knoll, located in the southern Ryukyu Arc, is known to actively bubble with gas containing methane and hydrogen sulfide from numerous fissures in the large carbonate pavement. Although ecological studies regarding macrobenthos and bacteria from Kuroshima Knoll have been intensively conducted, the community structure and ecological importance of microbial eukaryotes (protists) have not yet been investigated. In the present study, we directly extracted DNA from sediment of the Kuroshima Knoll at a depth of 640 m and constructed genetic libraries of PCR-amplified eukaryotic small-subunit ribosomal DNA (SSU rDNA). Although the SSU rDNA sequences of several types of benthic foraminifers were retrieved from the surface of the sediment, all other sequences (just below the sediment surface to approximately 9 cm below sediment surface) were derived from the basidiomycetous yeast Cryptococcus curvatus. Furthermore, sequences of the internal transcribed spacer of rDNA (ITS-rDNA) retrieved from the same sediment were identical to that of C. curvatus originating from terrestrial habitats. The diversity of microbial eukaryotes in the Kuroshima Knoll sediment seems to be extremely low and significantly different from that of other marine environments previously reported. PMID- 16341820 TI - A highly thermostable ferritin from the hyperthermophilic archaeal anaerobe Pyrococcus furiosus. AB - A ferritin from the obligate anaerobe and hyperthermophilic archaeon Pyrococcus furiosus (optimal growth at 100 degrees C) has been cloned and overproduced in Escherichia coli to one-fourth of total cell-free extract protein, and has been purified in one step to homogeneity. The ferritin (PfFtn) is structurally similar to known bacterial and eukaryal ferritins; it is a 24-mer of 20 kDa subunits, which add up to a total Mr 480 kDa. The protein belongs to the non-heme type of ferritins. The 24-mer contains approximately 17 Fe (as isolated), 2,700 Fe (fully loaded), or <1 Fe (apoprotein). Fe-loaded protein exhibits an EPR spectrum characteristic for superparamagnetic core formation. At 25 degrees C V(max) = 25 micromole core Fe(3+) formed per min per mg protein when measured at 315 nm, and the K(0.5) = 5 mM Fe(II). At 0.3 mM Fe(II) activity increases 100-fold from 25 to 85 degrees C. The wild-type ferritin is detected in P. furiosus grown on starch. PfFtn is extremely thermostable; its activity has a half-life of 48 h at 100 degrees C and 85 min at 120 degrees C. No apparent melting temperature was found up to 120 degrees C. The extreme thermostability of PfFtn has potential value for biotechnological applications. PMID- 16341821 TI - Transfer of siRNA against XIAP induces apoptosis and reduces tumor cells growth potential in human breast cancer in vitro and in vivo. AB - BACKGROUND: Gene targeting using short interfering RNA(siRNA) has become a common strategy to explore gene function because of its prominent efficacy and specificity. It is proven that the application of siRNA technology to gene therapy is effective. In this study, we constructed a siRNA expression plasmid against gene X-linked inhibitor of apoptosis (XIAP), and then used breast cancer cells MCF-7 to assess its functions. MATERIALS AND METHODS: XIAP siRNA plasmid was constructed using an U6pro vector contained U6 promoter, After the plasmid had been transfected into MCF-7 cells and effected on the cell cycle, the expression change of XIAP was examined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The apoptosis of the transfected cells was analyzed by flow cytometry, and TUNEL method. The in vitro cellular growth activities were assayed by MTT incorporation. Twenty-four nude mice were randomly divided into 3 equal groups and were inoculated with electroinjection of blank plasmid, scrambled nucleotide control (control siRNA), or siRNA against XIAP subcutaneously respectively, then the appearance and size of tumors were observed. Four weeks later the mice were killed and the volumes of tumor were calculated so as to evaluate the therapeutic effects of siRNA against XIAP. RESULTS: The successful construction of siRNA against XIAP plasmid was identified with sequencing. After the siRNA expression vector was transfected into the MCF-7 cells, the expression of XIAP gene was inhibited significantly (by 90%). The cellular growth activities in the MCF-7 cells transfected with siRNA against XIAP plasmid decreased obviously. The siRNA against XIAP plasmid knocked down XIAP expression in MCF-7 cells obviously, arrested the cell cycle in G1 phase, inhibited cell proliferation significantly, and promoted cell apoptosis in a tendency. TUNEL assay and flow cytometry showed that the classic apoptosis characters of the MCF-7 cells transfected with siRNA against XIAP plasmid manifested an apoptosis rate of 77.2%, significantly higher than those in the control siRNA group and in the blank plasmid group (both p < 0.01). The growth speed and formation rate of xenograft tumor in mice transfected with siRNA against XIAP transfected mice slowed down significantly. By HE staining, a lot of necrotic tissues could be observed in the siRNA against XIAP transfected group, however, there was no similar inhibitive effect in the control siRNA or blank plasmid group. CONCLUSION: This study represents that MCF-7 transfected cells with siRNA against XIAP remarkably suppress tumor growth and induces apoptosis, both in vitro and in vivo. This novel modality may be a promising tool for cancer therapy. PMID- 16341822 TI - Stimulus over-selectivity in rats. AB - The present study explored whether a similar phenomenon to stimulus over selectivity occurred in rats, in the hope of establishing a non-human model for the autism. Rats were serially presented with two-15 seconds, two-element compound stimuli prior to the delivery of food, in an appetitive classical conditioning procedure. Each compound stimulus consisted of two lights. Once the rats had acquired a conditioned response (CR) to the stimuli, they were presented with each of the component elements separately in extinction. The rats demonstrated greater conditioning to components of the compound presented just prior to reinforcement than to the components of the temporally distant compound. However, there was a smaller difference between CRs to the components of the compound presented just prior to reinforcement (i.e. less overshadowing) than between the components of the temporally distant compound. It is suggested that rats demonstrated a form of stimulus over-selectivity, resulting in greater overshadowing of one cue by another. Such results may form the basis of a viable non-human model of this symptom of autistic spectrum disorder. PMID- 16341823 TI - Crossed retroperitoneal approach to the internal iliac artery: a preliminary anatomical study. AB - The internal iliac arteries (IIA), and especially the distal segment, course very deep in the pelvis and are generally difficult to access surgically. The recent development of simple and reliable methods to investigate proximal ischemia of the lower extremities has led to discovery of more candidates for elective revascularization of the IAA. The classic approaches to the IAA, i.e., the transperitoneal and homolateral retroperitoneal routs have certain disadvantages. We present a new crossed retroperitoneal approach to the IAA and the results of such method in six cadavers. In all six cases, the entire IAA could be exposed without any particular problems. This novel approach to the IAA seems to be both simple and reproducible. PMID- 16341824 TI - An assessment on spiral CT scan of the superior and inferior genial spinal foramina and canals. AB - To characterise the superior as well as the inferior genial spinal foramen and their bony canal using a large CT data sample. The study included 555 mandibular spirals CT scans, taken from patients for pre-operative implant planning. Basic observation by three observers included the number of canals, their respective anatomical location and morphological variations. Furthermore, linear measurements were performed to describe the foramina and canal characteristics. On spiral CT scans, the superior and inferior genial spinal foramina were detected in 448 CT scans (81%). In 29% of the mandibular CTs, two or more midline canals were noted. 47% of the foramina were located in a position inferior to the genial spines; the others were located in a superior position. The mean (SD) lingual and buccal diameters of the canal structures were 0.73 (0.27) and 0.54 (0.29) mm, respectively with an average length of 3.94 (1.29) mm. The canal was located on average 7.40 (5.31) mm from the mandibular base at the lingual entrance and 7.96 (3.47) mm at the buccal canal end point. The average canal extended towards the buccal bone for an average 37% of the bone width. The results demonstrated the presence of both superior and inferior genial spinal foramina, with 29% of the individuals having double foramina. About 53% of the foramina was located superior to the genial spines. Considering its neurovascular content, these foramina should be carefully evaluated during pre-operative planning. PMID- 16341825 TI - Assessment of four midcarpal radiologic determinations. AB - Several radiologic measurement methods have been described for determining static carpal alignment of the wrist. These include the scapholunate, radiolunate, and capitolunate angles. The triangulation method is an alternative radiologic measurement which we believe is easier to use and more reproducible and reliable than the above mentioned methods. The purpose of this study is to assess the intraobserver reproducibility and interobserver reliability of the triangulation method, scapholunate, radiolunate, and capitolunate angles. Twenty orthopaedic residents and staff at varying levels of training made four radiologic measurements including the scapholunate, radiolunate and capitolunate angles as well as the triangulation method on five different lateral, digitized radiographs of the wrist and forearm in neutral radioulnar deviation. Thirty days after the initial measurements, the participants repeated the four radiologic measurements using the same radiographs. The triangulation method had the best intra-and interobserver agreement of the four methods tested. This agreement was significantly better than the capitolunate and radiolunate angles. The scapholunate angle had the next best intraobserver reproducibility and interobserver reliability. The triangulation method has the best overall observer agreement when compared to the scapholunate, radiolunate, and capitolunate angles in determining static midcarpal alignment. No comment can be made on the validity of the measurements since there is no radiographic gold standard in determining static carpal alignment. PMID- 16341826 TI - Burnout in the general population. Results from the Finnish Health 2000 Study. AB - BACKGROUND: Burnout is a chronic stress syndrome which develops gradually as a consequence of prolonged stress situation. Socio-demographic factors related to job-related burnout have not been studied in the whole population. We investigated the relative differences in the level of burnout between groups based on various socio-demographic factors in the population-based Finnish sample. METHODS: The nationally representative sample comprised 3,424 employees aged 30-64 years. Burnout was assessed with the Maslach Burnout Inventory-General Survey. The socio-demographic factors of interest were gender, age, education, type of employment, work experience, socio-economic status (SES), working time, and marital status. RESULTS: Only small differences in burnout were found between the different population groups. As a three-dimensional syndrome, burnout was associated with age. In contrast to what has been consistently reported so far, mostly among human service work and in non-representative studies, burnout seemed to increase somewhat with age. Among women, burnout was also related to education, SES, and work experience, and among men, to marital status. CONCLUSIONS: Burnout can evolve in all kinds of vocational groups. It seems that age does not generally protect against burnout. A low education level and low social status carry a possible risk of burnout for women, and being single, divorced, or widowed carry a possible risk of burnout for men. PMID- 16341828 TI - Use of mental health services in a developing country. Results from the Nigerian survey of mental health and well-being. AB - BACKGROUND: Evidence from developed industrialized countries suggests poor uptake of mental health services. No data exist in developing resource-constrained countries about met and unmet need for mental health service in the community. METHOD: A four-stage stratified probability sample of households was studied in the Yoruba-speaking part of Nigeria (population, approximately 25 million people or 22% of the Nigerian national population). Face-to-face interviews were conducted with persons 18 years old and above (n=4,984) using the World Mental Health version of the Composite International Diagnostic Interview. We determined the proportions of respondents with 12-month Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) anxiety, mood, or substance use disorder who had received any mental health treatment and the correlates of treatment receipt. RESULTS: Only 9.0% of those with any 12-month DSM-IV disorder had received treatment. While 11% of those with a mood disorder had received some treatment, none of those with substance use disorders had used a mental health service. Most treatments were received from general medical settings, with only about 1% of those with DSM-IV disorders receiving specialist mental health service. Surprisingly, complementary or alternative health providers were also consulted by only about 4% of those with mental disorders, although a much higher proportion of 57% of those with no DSM-IV disorders but who nevertheless received mental health treatment did so from such providers. Irrespective of the disorders or the sector where treatment was received, virtually no treatment was adjudged minimally adequate. CONCLUSION: There is a striking level of unmet need for mental health service in the community in this developing country setting. While inadequacy of the formal public health sector may be partly responsible for this observation, there is the likelihood that receipt of treatment for mental health problems may also be hampered by the public's poor knowledge of the nature of the disorders and by stigma. PMID- 16341827 TI - Understanding suicide in Australian farmers. AB - OBJECTIVE: Male farmers in Australia have an elevated risk of suicide. The aims of this study were to investigate the rate of mental health problems amongst farmers compared with non-farmer rural residents and to investigate what additional factors might contribute to an increased risk of suicide amongst farmers. METHOD: This study used a combination of quantitative and qualitative approaches. First, using self-report questionnaire data, we compared rates of mental health problems (a common correlate of suicide) and a number of personality measures between farmers (n=371) and non-farming rural residents (n=380). In addition, semi-structured interviews with farmers (n=32) were used to gain a richer understanding of how the context of farming and mental health interact. RESULTS: Five key findings emerged from the study. First, in the quantitative study, we found no support for the proposition that farmers experience higher rates of mental health problems than do non-farmer rural residents, but we identified potentially important personality differences between farmers and non-farmers, with levels of conscientiousness being significantly higher amongst farmers and levels of neuroticism being significantly lower. A strong association between maleness and farming was also found. In the qualitative study, participants indicated that farming is an environment in which individuals experienced a range of stressors but have limited capacity to acknowledge or express these. In addition, there appeared to be significant attitudinal barriers to seeking help for those who may have mental health problems, particularly male farmers. CONCLUSION: The elevated rate of suicide amongst farmers does not seem to be simply explained by an elevated rate of mental health problems. Individual personality, gender and community attitudes that limit a person's ability to acknowledge or express mental health problems and seek help for these may be significant risk factors for suicide in farmers. PMID- 16341829 TI - The prevalence of nursing staff stress on adult acute psychiatric in-patient wards. A systematic review. AB - BACKGROUND: Concerns about recent changes in acute in-patient mental health care environments have led to fears about staff stress and poor morale in acute in patient mental health care staff. AIM: To review the prevalence of low staff morale, stress, burnout, job satisfaction and psychological well-being amongst staff working in in-patient psychiatric wards. METHOD: Systematic review. RESULTS: Of 34 mental health studies identified, 13 were specific to acute in patient settings, and 21 were specific to other non-specified ward-based samples. Most studies did not find very high levels of staff burnout and poor morale but were mostly small, of poor quality and provided incomplete or non-standardised prevalence data. CONCLUSIONS: The prevalence of indicators of low morale on acute in-patient mental health wards has been poorly researched and remains unclear. Multi-site, prospective epidemiological studies using validated measures of stress together with personal and organizational variables influencing staff stress in acute in-patient wards are required. PMID- 16341830 TI - Poly-Ig tandems from I-band titin share extended domain arrangements irrespective of the distinct features of their modular constituents. AB - The cellular function of the giant protein titin in striated muscle is a major focus of scientific attention. Particularly, its role in passive mechanics has been extensively investigated. In strong contrast, the structural details of this filament are very poorly understood. To date, only a handful of atomic models from single domain components have become available and data on poly-constructs are limited to scarce SAXS analyses. In this study, we examine the molecular parameters of poly-Ig tandems from I-band titin relevant to muscle elasticity. We revisit conservation patterns in domain and linker sequences of I-band modules and interpret these in the light of available atomic structures of Ig domains from muscle proteins. The emphasis is placed on features expected to affect inter domain arrangements. We examine the overall conformation of a 6Ig fragment, I65 I70, from the skeletal I-band of soleus titin using SAXS and electron microscopy approaches. The possible effect of highly conserved glutamate groups at the linkers as well as the ionic strength of the medium on the overall molecular parameters of this sample is investigated. Our findings indicate that poly-Ig tandems from I-band titin tend to adopt extended arrangements with low or moderate intrinsic flexibility, independently of the specific features of linkers or component Ig domains across constitutively- and differentially-expressed tandems. Linkers do not appear to operate as free hinges so that lateral association of Ig domains must occur infrequently in samples in solution, even that inter-domain sequences of 4-5 residues length would well accommodate such geometry. It can be expected that this principle is generally applicable to all Ig-tandems from I-band titin. PMID- 16341831 TI - Patchiness and spatial distribution of laccase genes of ectomycorrhizal, saprotrophic, and unknown basidiomycetes in the upper horizons of a mixed forest cambisol. AB - Decomposition of plant litter by the soil microbial community is an important process of controlling nutrient cycling and soil humus formation. Fungal laccases are key players in litter-associated polyphenol degradation, but little is known about the diversity and spatial distribution of fungal species with laccase genes in soils. Diversity of basidiomycete laccase genes was assessed in a cambisolic forest soil, and the spatial distribution of the sequences was mapped in a 100 m(2) plot by using polymerase chain reaction (PCR) on soil DNA extracts. Diversity of laccase sequences was higher in the organic horizon and decreased with the depth. A total of 167 different sequences sharing 44-96% oligonucleotide similarity was found in 13 soil cores harvested in the 100-m(2) plot. Dissimilarity in laccase sequence content was 67% between adjacent cores; 45.5%, 35.5% and 19% of laccase sequences were attributed to ectomycorrhizal, unknown and saprotrophic basidiomycetes, respectively. Most dominant sequences were attributed to the extramatrical hyphae of known ectomycorrhizal taxa (e.g., Russulaceae) and restricted to small patches (<0.77 m(2)) in a specific soil horizon. Soil fungi with laccase genes occupied different niches and showed strikingly variable distribution patterns. The distribution of laccase sequences, and corresponding fungi, likely reflected a part of the oxidative potential in soils. PMID- 16341832 TI - [Clinical interdisciplinary gastroenterology]. PMID- 16341833 TI - First performance evaluation of a dual-source CT (DSCT) system. AB - We present a performance evaluation of a recently introduced dual-source computed tomography (DSCT) system equipped with two X-ray tubes and two corresponding detectors, mounted onto the rotating gantry with an angular offset of 90 degrees . We introduce the system concept and derive its consequences and potential benefits for electrocardiograph [corrected] (ECG)-controlled cardiac CT and for general radiology applications. We evaluate both temporal and spatial resolution by means of phantom scans. We present first patient scans to illustrate the performance of DSCT for ECG-gated cardiac imaging, and we demonstrate first results using a dual-energy acquisition mode. Using ECG-gated single-segment reconstruction, the DSCT system provides 83 ms temporal resolution independent of the patient's heart rate for coronary CT angiography (CTA) and evaluation of basic functional parameters. With dual-segment reconstruction, the mean temporal resolution is 60 ms (minimum temporal resolution 42 ms) for advanced functional evaluation. The z-flying focal spot technique implemented in the evaluated DSCT system allows 0.4 mm cylinders to be resolved at all heart rates. First clinical experience shows a considerably increased robustness for the imaging of patients with high heart rates. As a potential application of the dual-energy acquisition mode, the automatic separation of bones and iodine-filled vessels is demonstrated. PMID- 16341834 TI - [Seasonal fluctuations and influence of nutrition on macular pigment density]. AB - PURPOSE: We have previously reported on measuring macular pigment density (MPD) with a scanning laser ophthalmoscope (HRA, Heidelberg Engineering, Heidelberg, Germany). This study war undertaken to evaluate the variation of MPD over a period of 1 year in healthy subjects. METHOD: We used autofluorescence images recorded with a HRA to evaluate MPD with a 2 degrees circle centered on the fovea. Healthy subjects were included in the study and MPD measurements were repeated every 2 months over a period of 1 year. RESULTS: We included a total of 30 healthy subjects aged 19-34 years (mean: 23+/-2 years). Mean MPD at time point 1 was 0.215+/-0.056 density units (DU), at time point 2 0.235+/-0.051 DU, at time point 3 0.218+/-0.055 DU, at time point 4 0.228+/-0.057 DU, at time point 5 0.225+/-0.053 DU, and at time point 6 0.203+/-0.050 DU. The statistical analysis revealed no significant variation of MPD over the follow-up period of 1 year. CONCLUSION: This study demonstrates that MPD shows no variation over a period of 1 year in healthy subjects. PMID- 16341835 TI - Torularhodin and torulene are the major contributors to the carotenoid pool of marine Rhodosporidium babjevae (Golubev). AB - A carotenoid-producing yeast strain, isolated from the sub-arctic, marine copepod Calanus finmarchicus, was identified as Rhodosporidium babjevae (Golubev) according to morphological and biochemical characteristics and phylogenetic inference from the small-subunit ribosomal RNA gene sequence. The total carotenoids content varied with cultivation conditions in the range 66-117 microg per g dry weight. The carotenoid pool, here determined for the first time, was dominated by torularhodin and torulene, which collectively constituted 75-91% of total carotenoids under various regimes of growth. Beta-carotene varied in the range 5-23%. A high-peptone/low-yeast extract (weight ratio 38:1) marine growth medium favoured the production of torularhodin, the carotenoid at highest oxidation level, with an average of 63% of total carotenoids. In standard yeast medium (YM; ratio 1.7:1), torularhodin averaged 44%, with increased proportions of the carotenes, torulene and beta-carotene. The anticipated metabolic precursor gamma-carotene (beta,psi-carotene) constituted a minor fraction (60 s. An application of mobile, membrane-permeant pH buffers accelerated the collapse of the alkaline-pH gradients but, even at 30 mM, was unable to abolish them. Modeling of the pH(i) dynamics showed that both the relatively weak effect of the weak acid/base on the peak size of the pH gradient and the accelerated collapse of the pH gradient could be due to the time taken for equilibration of the weak acid and base across the cell. We propose that appropriate weak acid/base mixes may provide a simple method for studying the role of local pH(i) signals without perturbing steady state pH(i). Furthermore, an extrapolation of our in vitro data to longer and thinner neuronal structures found in the mammalian nervous system suggests that dendritic and axonal pH(i) are likely to be dominated by local pH(i)-regulating mechanisms rather than simply following the soma pH(i). PMID- 16341878 TI - Cloning and analysis of a cDNA encoding a putative serine protease comprising two trypsin-like domains of Trichinella spiralis. AB - The cDNA encoding a putative serine protease, TsSerP, was cloned by degenerative polymerase chain reaction and screening of the cDNA library from Trichinella spiralis adult-newborn larvae stage. Sequence analysis revealed the presence of two trypsin-like serine protease domains flanking a hydrophilic domain, with the catalytic triad residue histidine in the alpha domain substituted by an arginine residue. Southern blots indicated that this was a single copy gene in the parasite genome. Northern blots demonstrated a single 2.3-kb transcript during the muscle larvae and adult stages of T. spiralis. The recombinant protein from the TsSerP beta domain (betaSerP) was produced but not recognised by T. spiralis infected swine serum. An anti-betaSerP polyclonal serum detected a 69-kDa polypeptide in the soluble antigens of T. spiralis muscle larvae. Immunolocalisation analysis located TsSerP on the inner layer of the cuticle and oesophagus of the parasite, suggesting a potential role in its moulting and/or digestive functions. PMID- 16341879 TI - Assessment of a liposomal formulation of ivermectin in rabbit after a single subcutaneous administration. AB - Ivermectin is a member of the macrocyclic lactone family widely used in livestock, pets, and humans as a potent parasiticide. Slight differences in formulation may change the plasma kinetics and efficacy of these compounds. The aim of the study is to evaluate the ability of a liposomal formulation of ivermectin to generate an efficient exposure of the animal to the drug. Ten rabbits were subcutaneously administered with 0.3 mg kg(-1) of ivermectin using Ivomec (n=5) or a liposomal formulation (n=5). The areas under serum concentration-time curve were similar after both treatments, indicating the same bioavailability for the two formulations. However, the liposomal formulation gave a higher C(max) value (33.33 ng ml(-1)) compared with Ivomec (20.82 ng ml(-1)) and a significantly faster absorption as indicated by the T(max) of 0.23 days compared with 1.13 days for the Ivomec formulation. The use of liposomal formulation shows promise as this system improves the efficacy of ivermectin and related drugs. PMID- 16341880 TI - In vitro evaluation of trypanocidal activity in plants used in Argentine traditional medicine. AB - Thirty-two organic and aqueous extracts, belonging to 12 Argentine medicinal plants were tested for their in vitro trypanocidal activity on epimastigote forms of Trypanosoma cruzi. Among the selected species, the organic extracts of Ambrosia scabra, Ambrosia tenuifolia, Baccharis spicata, Eupatorium buniifolium, Lippia integrifolia, Mulinum spinosum and Satureja parvifolia, and the aqueous extracts of E. buniifolium, L. integrifolia, M. spinosum and S. parvifolia showed trypanocidal activity with a percentage of growth inhibition higher than 70% at a concentration of 100 microg/ml. PMID- 16341882 TI - The reproductive success of the parasitic bat fly Basilia nana (Diptera: Nycteribiidae) is affected by the low roost fidelity of its host, the Bechstein's bat (Myotis bechsteinii). AB - We studied the reproductive ecology of the bat fly Basilia nana on free-ranging colonial female and solitary male Bechstein's bats (Myotis bechsteinii) during one reproductive season. The reproduction of B. nana took place from April to September, and the production of puparia in bat roosts was high. The metamorphosis of the flies took a minimum of 30 days, and at least 86% of the puparia metamorphosed successfully. However, only about 30% of flies from puparia deposited in female roosts and 57% of flies from puparia deposited in male roosts emerged in the presence of Bechstein's bats and were thus able to survive. The significantly higher emergence success of bat flies in male roosts was caused by the higher roost fidelity of the solitary males compared with the social females. Our results indicate that bats can control the reproductive success of bat flies by switching and selecting roosts. PMID- 16341881 TI - Cloning and characterization of the Cu/Zn superoxide dismutase of Trichinella pseudospiralis. AB - Copper/zinc (Cu/Zn) superoxide dismutase (SOD) activity was identified for the first time in both crude somatic extracts (CE) and excretory/secretory (E/S) products of Trichinella pseudospiralis. It was the dominant SOD in infective stage larvae. Native polyacrylamide gel electrophoresis of CE and E/S products yielded a prominent band, which was cyanide-sensitive and was partly inhibited by hydrogen peroxide in SOD assay. Cytosolic Cu/Zn SOD was cloned. The 471-bp full length cDNA sequence contained an open reading frame of 157 amino acids. The gene contained three introns. Quantitative reverse transcription-polymerase chain reaction indicated that the expression of cytosolic Cu/Zn SOD was substantially higher in infective-stage larvae than in adult worms. Cluster analysis showed that the sequence of the Cu/Zn SOD of T. pseudospiralis, an adenophorean nematode, is related to those of Brugia pahangi, Acanthocheilonema viteae, Onchocerca volvulus, and Haemonchus contortus (all belonging to the sercenentean group). PMID- 16341883 TI - Could the homologous sequence of anti-inflammatory pentapeptide (MLIF) produced by Entamoeba histolytica in the N protein of rabies virus affect the inflammatory process? AB - Amebiasis and rabies are public health problems, and they have in common a poor inflammatory effect in the target organs that they affect. In the GenBank, it was found that the anti-inflammatory peptide monocyte locomotion inhibitory factor (MLIF) produced by Entamoeba histolytica homologates 80%, with a fragment of the N protein of the rabies virus. We speculated if the N protein could contribute to the scant inflammatory reaction produced by rabies virus in central nervous system. The N protein was obtained and studied in vitro and in vivo. The N protein, as MLIF, inhibited the respiratory burst in human mononuclear phagocytes (43%, p<0.05), but in contrast to MLIF, it increased chemotaxis and it did not significantly inhibit delayed hypersensitivity skin reaction to 1-chloro-2-4 dinitrobenzene in guinea pigs. Therefore, the full peptide sequence has to be present or it has to be cleaved-free from the large recombinant N protein molecule (55 kDa) to become active. PMID- 16341886 TI - Changes in delta 13C stable isotopes in multiple tissues of insect predators fed isotopically distinct prey. AB - Traditionally, researchers have used measurements of carbon stable isotopes to infer the composition of consumers' diets. However, since consumer's tissues may process carbon isotopes differently, particularly following a diet shift, it is possible to use measurements of carbon isotopes in multiple tissues to determine not only the composition of an individual's diet, but also the temporal dynamics thereof. This study examined how stable isotopes of carbon (13C/12C, expressed as delta 13C) changed in different adult tissues of two predacious beetles, Harmonia axyridis and Coccinella septempunctata (Coleoptera: Coccinellidae). In the laboratory, we switched ladybeetles from a C3-based diet (soybean aphids, Aphis glycines) to a C4-based one (corn leaf aphids, Rhopalosiphum maidis). The delta 13C of metabolically active tissues such as the body fat and reproductive organs changed rapidly (< or =5 days) following the diet shift. Tissues expected to be more metabolically inert, such as wings, changed more slowly over the same period. Although these general patterns were largely similar between males and females, females had more rapid changes in delta 13C in fat and reproductive tissues. However, females showed a significant depletion in delta 13C after 10 days, while males' delta 13C continued to increase. Given the results of this experiment, it is now possible to distinguish between ladybeetles eating a mixed diet (beetles with multiple tissues at similar, intermediate, equilibrial delta 13C signatures) from those that have shifted diets (beetles with different tissues at distinctly different delta 13C values). Thus, this approach can be used broadly to infer not only what constitutes the diet of a consumer, but also the temporal history of dietary intake. PMID- 16341884 TI - Recruitment of RecA homologs Dmc1p and Rad51p to the double-strand break repair site initiated by meiosis-specific endonuclease VDE (PI-SceI). AB - During meiosis, VDE (PI-SceI), a homing endonuclease in Saccharomyces cerevisiae, introduces a double-strand break (DSB) at its recognition sequence and induces homologous recombinational repair, called homing. Meiosis-specific RecA homolog Dmc1p, as well as mitotic RecA homolog Rad51p, acts in the process of meiotic recombination, being required for strand invasion and exchange. In this study, recruitment of Dmc1p and Rad51p to the VDE-induced DSB repair site is investigated by chromatin immunoprecipitation assay. It is revealed that Dmc1p and Rad51p are loaded to the repair site in an independent manner. Association of Rad51p requires other DSB repair proteins of Rad52p, Rad55p, and Rad57p, while loading of Dmc1p is facilitated by the different protein, Sae3p. Absence of Tid1p, which can bind both RecA homologs, appears specifically to cause an abnormal distribution of Dmc1p. Lack of Hop2, Mnd1p, and Sae1p does not impair recruitment of both RecA homologs. These findings reveal the discrete functions of each strand invasion protein in VDE-initiated homing, confirm the similarity between VDE-initiated homing and Spo11p-initiated meiotic recombination, and demonstrate the availability of VDE-initiated homing for the study of meiotic recombination. PMID- 16341885 TI - Barley necrotic locus nec1 encodes the cyclic nucleotide-gated ion channel 4 homologous to the Arabidopsis HLM1. AB - Barley homolog of the Arabidopsis necrotic (disease lesion mimic) mutant HLM1 that encodes the cyclic nucleotide-gated ion channel 4 was cloned. Barley gene was mapped genetically to the known necrotic locus nec1 and subsequent sequence analysis identified mutations in five available nec1 alleles confirming barley homolog of Arabidopsis HLM1 as the NEC1 gene. Two fast neutron (FN) induced mutants had extensive deletions in the gene, while two previously described nec1 alleles had either a STOP codon in exon 1 or a MITE insertion in intron 2 which caused alternative splicing, frame shift and production of a predicted non functional protein. The MITE insertion was consistent with the reported spontaneous origin of the nec1 Parkland allele. The third FN mutant had a point mutation in the coding sequence which resulted in an amino acid change in the conserved predicted cyclic nucleotide-gated ion channel pore region. The expression of two pathogenesis-related genes, HvPR-1a and beta-1,3-glucanase, was elevated in two FN necrotic lines. Ten other members of the barley cyclic nucleotide-gated ion channel gene family were identified and their position on barley linkage map is reported. PMID- 16341887 TI - Cladocerans versus copepods: the cause of contrasting top-down controls on freshwater and marine phytoplankton. AB - Top-down control of phytoplankton by crustacean mesozooplankton is a cornerstone of freshwater ecology. Apparently, trophic cascades are more frequently reported from freshwater than from marine plankton. We argue that this difference is real and mainly caused by biological differences at the zooplankton-phytoplankton link: cladocerans (particularly Daphnia) in the lakes and copepods in the sea. We derive these conclusions from recent literature and a number of own, similarly designed mesocosm experiments conducted in a lake, a brackish water and a marine site. In all experiments, phytoplankton were exposed to gradients of experimentally manipulated densities of zooplankton, including freshwater copepods and cladocerans, and marine copepods and appendicularians. The suggested reasons for the difference between lake and marine trophic cascades are: (1) Both copepods and cladocerans suppress only part of the phytoplankton size spectrum: cladocerans the small and copepods the large phytoplankton. (2) If not controlled by grazing, small phytoplankton may increase their biomass faster than large phytoplankton. (3) Copepods additionally release small phytoplankton from grazing pressure by intermediate consumers (protozoa) and competitors (predation on appendicularian eggs), while cladocerans do not release large phytoplankton from grazing pressure by any functional group. (4) Cladocerans sequester more of the limiting nutrient than copepods, leaving fewer nutrients available for compensatory growth of ungrazed phytoplankton. PMID- 16341888 TI - Carotenoids modulate the trade-off between egg production and resistance to oxidative stress in zebra finches. AB - The allocation of resources to reproduction and survival is a central question of studies of life history evolution. Usually, increased allocation to current reproduction is paid in terms of reduced future reproduction and/or decreased survival. However, the proximal mechanisms underlying the cost of reproduction are poorly understood. Recently, it has been shown that increased susceptibility to oxidative stress might be one of such proximate links between reproduction and self-maintenance. Organisms possess a range of antioxidant defenses, including endogenously produced molecules (e.g., enzymes) and compounds ingested with food (e.g., carotenoids). If reproductive effort increases the production of reactive oxygen species, the availability of antioxidant defenses may partly or fully counteract the free-radical damages. One could, therefore, expect that the trade off between reproduction and oxidative stress is modulated by the availability of antioxidant defenses. We tested this hypothesis in zebra finches. We manipulated reproductive effort by either allowing or preventing pairs to breed. Within each breeding or non-breeding group, the availability of antioxidant compounds was manipulated by supplementing or not supplementing the drinking water with carotenoids. We found that although birds in the breeding and non-breeding groups did not differ in their resistance to oxidative stress (the breakdown of red blood cells submitted to a controlled free-radical attack), one aspect of breeding effort (i.e., the number of eggs laid by birds in both breeding and non breeding groups) was negatively correlated with resistance to oxidative stress only in birds that did not benefit from a carotenoid-supplemented diet. This result therefore suggests that carotenoid availability can modulate the trade-off between reproduction and resistance to oxidative stress. PMID- 16341889 TI - Spatial synchrony of prairie ducks: roles of wetland abundance, distance, and agricultural cover. AB - Populations exhibit spatial synchrony when their numbers rise and fall in concert at several sites over their distribution. I examined the relationship between synchrony, abundance of wetlands (ponds), distance, and agricultural cover using count data of ten duck species counted in 23 aerial survey strata on the mid continental prairies of North America. Expansion of agriculture may have resulted in increased synchrony of duck populations through increased foraging efficiency of nomadic predators and/or if the homogenization of nesting habitat has removed habitat features that allow differential local responses to large-scale population drivers such as precipitation. As a measure of synchrony, I calculated all pair-wise cross-correlation coefficients based on population growth rates (rt) at each survey stratum, and then regressed these correlation coefficients against measures of cross-correlation of pond (wetland) counts, distance between strata, and mean percent area of strata seeded to row crops. Synchrony for most species was most strongly related to synchrony of wetland availability among sites, and decreased with distance between sites. Synchrony of ducks that nest over water showed little effect of agricultural cover, whereas the effect of agricultural cover on synchrony of upland nesting ducks differed by species. Mobile large-bodied species showed evidence of increased synchrony due to agricultural cover, whereas smaller-bodied, more philopatric species showed evidence of decreased synchrony due to agricultural cover. PMID- 16341890 TI - Variation in predation pressure as a mechanism underlying differences in numerical abundance between populations of the poeciliid fish Heterandria formosa. AB - We explored whether a variation in predation and habitat complexity between conspecific populations can drive qualitatively different numerical dynamics in those populations. We considered two disjunct populations of the least killifish, Heterandria formosa, that exhibit long-term differences in density, top fish predator species, and dominant aquatic vegetation. Monthly censuses over a 3-year period found that in the higher density population, changes in H. formosa density exhibited a strong negative autocorrelation structure: increases (decreases) at one census tended to be followed by decreases (increases) at the next one. However, no such correlation was present in the lower density population. Monthly census data also revealed that predators, especially Lepomis sp., were considerably more abundant at the site with lower H. formosa densities. Experimental studies showed that the predation by Lepomis gulosus occurred at a much higher rate than predation by two other fish and two dragonfly species, although L. gulosus and L. punctatus had similar predation rates when the amount of vegetative cover was high. The most effective predator, L. gulosus, did not discriminate among life stages (males, females, and juveniles) of H. formosa. Increased predation rates by L. gulosus could keep H. formosa low in one population, thereby eliminating strong negative density-dependent regulation. In support of this, changes in H. formosa density were positively correlated with changes in vegetative cover for the population with a history of lower density, but not for the population with a history of higher density. Our results are consistent with the hypothesis that the observed differences among natural populations in numerical abundance and dynamics are caused in part by the differences in habitat complexity and the predator community. PMID- 16341891 TI - Nutritional differences and leaf acclimation of climbing plants and the associated vegetation in different types of an Andean montane rainforest. AB - Climbing plants are known to play an important role in tropical forest systems, but key features for their distribution are only partly understood. Investigation was carried out to find if climbers differ from self-supporting vegetation in their adjustment of leaf parameters over a wide variety of light regimes in different forest types along an altitudinal gradient. Relative photon flux density (PFDrel) was assessed above 75 pairs of strictly linked climbers and supporting vegetation on seven plots between 2,020 and 2,700 m a.s.l. along a mountain range in South-Ecuador up to the Paramo vegetation. Leaf samples from both growth forms were analyzed for leaf area (LA), specific leaf mass (LMA), mass and area-based carbon and nitrogen concentration (C, Carea, N, and Narea) and concentrations of P, K, Ca, Mg, Mn and Al. Leaf size of climbers was independent of general light condition, whereas the leaf size of the self supporting vegetation increased in shade. LMA increased as expected with altitude and irradiance for both growth forms, but climbers generally built smaller leaves with lower LMA. N, P, and K concentrations were higher in the leaves of climbers than in their supporters. Relationships of LMA and Narea to the light conditions were more pronounced within the climbers than within their supporters. Slope for the regression between climber's Narea and LMA was twice as steep as for the supporter leaves. Al accumulators were only found within the self-supporting vegetation. The investigated traits indicate improved adjustment towards light supply within climbers compared to self-supporting vegetation. Thus climbing plants seem to have a higher potential trade off in resource-use efficiency regarding irradiance and nutrients. PMID- 16341892 TI - Induced production of extrafloral nectar in intact lima bean plants in response to volatiles from spider mite-infested conspecific plants as a possible indirect defense against spider mites. AB - We found that intact lima bean plants increased the secretion of extrafloral nectar (EFN) after exposure to Tetranychus urticae-induced plant volatiles. Predatory mites, Phytoseiulus persimilis, dispersed more slowly from an exposed intact plant than from a control plant (plant exposed to volatiles from intact conspecific). The predators also dispersed more slowly from those plants that were provided with extra EFN than from untreated plants. We further show that EFN was a potential alternative food source for P. persimilis. From these results, we concluded that increased EFN was involved in the slow dispersal of P. persimilis from the plants exposed to herbivore-induced plant volatiles. Our data suggest that the increase of EFN in an HIPV-exposed intact plant could be an induced indirect defense against spider mites. PMID- 16341893 TI - Persistence of invading gypsy moth populations in the United States. AB - Exotic invasive species are a mounting threat to native biodiversity, and their effects are gaining more public attention as each new species is detected. Equally important are the dynamics of exotic invasives that are previously well established. While the literature reports many examples of the ability of a newly arrived exotic invader to persist prior to detection and population growth, we focused on the persistence dynamics of an established invader, the European gypsy moth (Lymantria dispar) in the United States. The spread of gypsy moth is largely thought to be the result of the growth and coalescence of isolated colonies in a transition zone ahead of the generally infested area. One important question is thus the ability of these isolated colonies to persist when subject to Allee effects and inimical stochastic events. We analyzed the US gypsy moth survey data and identified isolated colonies of gypsy moth using the local indicator of spatial autocorrelation. We then determined region-specific probabilities of colony persistence given the population abundance in the previous year and its relationship to a suite of ecological factors. We observed that colonies in Wisconsin, US, were significantly more likely to persist in the following year than in other geographic regions of the transition zone, and in all regions, the abundance of preferred host tree species and land use category did not appear to influence persistence. We propose that differences in region-specific rates of persistence may be attributed to Allee effects that are differentially expressed in space, and that the inclusion of geographically varying Allee effects into colony-invasion models may provide an improved paradigm for addressing the establishment and spread of gypsy moth and other invasive exotic species. PMID- 16341894 TI - Increased effect of harsh climate in red deer with a poor set of teeth. AB - Teeth are vital for mammal performance and especially in ungulates relying on mechanical decomposition of plant material for effective microbial digestion and energy uptake. The main focus of the role of teeth in ungulate life histories has been on tooth wear, while no one has addressed to what extent deviation from the natural set of teeth (maldentition) causes variation in individual fitness components. Based on mandibles from 41,066 individual red deer (Cervus elaphus L.) collected from 1969 to 2001, we tested whether maldentition had an effect on individual body condition and whether this effect depended on environmental harshness. Females with maldentition (0.6% of the population) were in a poorer condition than individuals without tooth anomalies and the effect increased during unfavorable climatic conditions. The effect of maldentition in males was less clear. This study indicates that a well-functioning set of teeth is essential for mammal performance, and that selection pressure against (dental) anomalies is more pronounced when climate is unfavorable. PMID- 16341895 TI - Organic and mineral fertilization, respectively, increase and decrease the development of external mycelium of arbuscular mycorrhizal fungi in a long-term field experiment. AB - Effects of long-term mineral fertilization and manuring on the biomass of arbuscular mycorrhizal fungi (AMF) were studied in a field experiment. Mineral fertilization reduced the growth of AMF, as estimated using both measurements of hyphal length and the signature fatty acid 16:1omega5, whereas manuring alone increased the growth of AMF. The results of AMF root colonization followed the same pattern as AMF hyphal length in soil samples, but not AMF spore densities, which increased with increasing mineral and organic fertilization. AMF spore counts and concentration of 16:1omega5 in soil did not correlate positively, suggesting that a significant portion of spores found in soil samples was dead. AMF hyphal length was not correlated with whole cell fatty acid (WCFA) 18:2omega6,9 levels, a biomarker of saprotrophic fungi, indicating that visual measurements of the AMF mycelium were not distorted by erroneous involvement of hyphae of saprotrophs. Our observations indicate that the measurement of WCFAs in soil is a useful research tool for providing information in the characterization of soil microflora. PMID- 16341896 TI - Resonance Raman fingerprinting of multiheme cytochromes from the cytochrome c3 family. AB - Resonance Raman (RR) spectroscopy was used to investigate conformational characteristics of the hemes of several ferricytochromes of the cytochrome c3 family, electron transfer proteins isolated from the periplasm and membranes of sulfate-reducing bacteria. Our analysis concentrated on the low-frequency region of the RR spectra, a fingerprint region that includes vibrations for heme-protein C-S bonds [nu(C(a)S)]. It has been proposed that these bonds are directly involved in the electron transfer process. The three groups of tetraheme cytochrome c3 analyzed, namely Type I cytochrome c (3) (TpIc (3)s), Type II cytochrome c (3) (TpIIc (3)s) and Desulfomicrobium cytochromes c3, display different frequency separations for the two nu(C(a)S) lines that are similar among members of each group. These spectral differences correlate with differences in protein structure observed among the three groups of cytochromes c3. Two larger cytochromes of the cytochrome c3 family display RR spectral characteristics for the nu(C(a)S) lines that are closer to TpIIc3 than to TpIc3. Two other multiheme cytochromes from Desulfovibrio that do not belong to the cytochrome c3 family display nu(C(a)S) lines with reverse relative areas in comparison with the latter family. This RR study shows that the small differences in protein structure observed among these cytochrome c3 correlate to differences on the heme-protein bonds, which are likely to have an impact upon the protein function, making RR spectroscopy a sensitive and useful tool for characterizing these cytochromes. PMID- 16341898 TI - The trans labilization of cis-[PtCl2(13CH3NH2)2] by glutathione can be monitored at physiological pH by [1H,13C] HSQC NMR. AB - In order to monitor the trans labilization of cisplatin at physiological pH we have prepared the complex cis-[PtCl(2)((13)CH(3)NH(2))(2)] and studied its interactions with excess glutathione in aqueous solution at neutral pH by two dimensional [1H,13C] heteronuclear single-quantum correlation (HSQC) NMR spectroscopy. [1H,13C] HSQC spectroscopy is a good method for following the release of (13)CH(3)NH(2) but is not so good for characterizing the Pt species in solution. In the reaction of cisplatin with glutathione, Pt-S bonds are formed and Pt-NH(3) bonds are broken. The best technique for following the formation of Pt-S bonds of cisplatin is by UV spectroscopy. [1H,13C] HSQC spectroscopy is the best method for following the breaking of the Pt-N bonds. [1H,15N] HSQC spectroscopy is the best method for characterizing the different species in solution. However, the intensity of the peaks in the (15)NH(3)-Pt-S region, in [1H,15N] HSQC, reflects a balance between the formation of Pt-S bonds, which increases the signal intensity, and the trans labilization, which decreases the signal intensity. [1H,15N] HSQC spectroscopy and [1H,13C] HSQC spectroscopy are complementary techniques that should be used in conjunction in order to obtain the most accurate information on the interaction of platinum complexes with sulfur-containing ligands. PMID- 16341897 TI - Structural evidence for a proton transfer pathway coupled with haem reduction of cytochrome c" from Methylophilus methylotrophus. AB - The crystal structures of the oxidized and reduced forms of cytochrome c" from Methylophilus methylotrophus were solved from X-ray synchrotron data to atomic resolution. The overall fold of the molecule in the two redox states is very similar and is comparable to that of the oxygen-binding protein from the purple phototrophic bacterium Rhodobacter sphaeroides. However, significant modifications occur near the haem group, in particular the detachment from axial binding of His95 observed upon reduction as well as the adoption of different conformations of some protonatable residues that form a possible proton path from the haem pocket to the protein surface. These changes are associated with the previously well characterized redox-Bohr behaviour of this protein. Furthermore they provide a model for one of the presently proposed mechanisms of proton translocation in the much more complex protein cytochrome c oxidase. PMID- 16341900 TI - pH-dependent redox and CO binding properties of chelated protoheme-L-histidine and protoheme-glycyl-L-histidine complexes. AB - The pH dependence of redox properties, spectroscopic features and CO binding kinetics for the chelated protohemin-6(7)-L-histidine methyl ester (heme-H) and the chelated protohemin-6(7)-glycyl-L-histidine methyl ester (heme-GH) systems has been investigated between pH 2.0 and 12.0. The two heme systems appear to be modulated by four protonating groups, tentatively identified as coordinated H(2)O, one of heme's propionates, N(epsilon) of the coordinating imidazole, and the carboxylate of the histidine residue upon hydrolysis of the methyl ester group (in acid medium). The pK (a) values are different for the two hemes, thus reflecting structural differences. In particular, the different strain at the Fe N(epsilon) bond, related to the different length of the coordinating arm, results in a dramatic alteration of the bond strength, which is much smaller in heme-H than in heme-GH. It leads to a variation in the variation of the pKa for the protonation of the N(epsilon) of the axial imidazole as well as in the proton linked behavior of the other protonating groups, envisaging a cross-talk communication mechanism among different groups of the heme, which can be operative and relevant also in the presence of the protein matrix. PMID- 16341899 TI - Recombination of the GFP gene to the BFP gene using a man-made site-selective DNA cutter. AB - By using the recently developed man-made DNA cutter [a combination of Ce(IV)/EDTA and two DNA additives], green fluorescent protein (GFP) was converted to closely related blue fluorescent protein (BFP). The phosphodiester linkages at T196-A200 in the sense strand of GFP were hydrolyzed by the cutter, and the A1-T196 fragment in the product was selectively connected with the downstream fragment (C197-A720) of BFP by T4 DNA ligase. This recombination changed three codons in the GFP gene (TGC at 196-198, TAT at 199-201, and ACC at 502-504) to TCT, CAT, and ATC in BFP, and accordingly three amino acids in GFP (Cys65, Tyr66, and Thr167) were altered to Ser65, His66, and Ile167. The recombinant gene was successfully expressed in Escherichia coli and emitted blue fluorescence, confirming the absence of undesired side reactions (mutation, deletion, insertion, depurination, etc.) in the DNA manipulation. PMID- 16341901 TI - Support-vector-machine classification of linear functional motifs in proteins. AB - Our algorithm predicts short linear functional motifs in proteins using only sequence information. Statistical models for short linear functional motifs in proteins are built using the database of short sequence fragments taken from proteins in the current release of the Swiss-Prot database. Those segments are confirmed by experiments to have single-residue post-translational modification. The sensitivities of the classification for various types of short linear motifs are in the range of 70%. The query protein sequence is dissected into short overlapping fragments. All segments are represented as vectors. Each vector is then classified by a machine learning algorithm (Support Vector Machine) as potentially modifiable or not. The resulting list of plausible post-translational sites in the query protein is returned to the user. We also present a study of the human protein kinase C family as a biological application of our method. PMID- 16341902 TI - Instruction-induced feature binding. AB - In order to test whether or not instructions specifying the stimulus-response (S R) mappings for a new task suffice to create bindings between specified stimulus and response features, we developed a dual task paradigm of the ABBA type in which participants saw new S-R instructions for the A-task in the beginning of each trial. Immediately after the A-task instructions, participants had to perform a logically independent B-task. The imperative stimulus for the A-task was presented after the B-task had been executed. The present data show that the instructed S-R mappings influence performance on the embedded B-task, even when they (1) have never been practiced, and (2) are irrelevant with respect to the B task. These results imply that instructions can induce bindings between S- and R features without prior execution of the task at hand. PMID- 16341904 TI - Morbidity and mortality of hospitalized patients with diffuse idiopathic skeletal hyperostosis. AB - Diffuse idiopathic skeletal hyperostosis (DISH) has been associated with various metabolic disorders considered to be cardiovascular risk factors such as obesity, diabetes mellitus, hyperinsulinemia, and hyperlipidemia. To evaluate morbidity and mortality of hospitalized patients with DISH admitted to the department of medicine. One hundred patients from a cohort of 1020 consecutive patients, aged 45 years and more, admitted to the department of medicine were diagnosed as suffering from DISH. Another group of 100 patients, age- and gender matched, admitted without DISH, served as controls. Clinical and demographic characteristics, diagnoses on admission, previous chronic diseases, chronic medical therapy, laboratory tests, and the rates of in-hospital mortality and readmissions within 1 month of discharge were collected from the hospital database, for the two groups. Uncompensated or paroxysmal atrial fibrillation was more often encountered on admission in patients with DISH (p = 0.038). Patients with DISH were more likely to suffer from elevated body mass index, arterial hypertension, diabetes mellitus, and previous cerebral vascular accidents, although the differences did not reach statistical significance. However, significantly more patients had an electrocardiographic evidence of left ventricular hypertrophy (p = 0.03). The mortality rate was similar between the two groups. The lack of significant associations for cardiovascular risk factors such as diabetes mellitus, hypertension, and high BMI should be interpreted cautiously considering the characteristics of the control group. Identification of comorbid conditions and proper therapeutic interventions may prove useful in reducing the morbidity and mortality associated with this disorder. PMID- 16341903 TI - Antineutrophil cytoplasmic antibodies in Bulgarian patients with rheumatoid arthritis: characterization and clinical associations. AB - OBJECTIVE: The aim of this study was to study the prevalence, subspecificities, and immunoglobulin (Ig)G subclass distribution of antineutrophil cytoplasmic antibodies (ANCA) in 90 Bulgarian patients with rheumatoid arthritis (RA) and to investigate the clinical associations of ANCA in these patients. METHODS: The ANCA were detected by indirect immunofluorescence, while antigen specificities were determined by enzyme-linked immunosorbent assay (ELISA) directed against myeloperoxidase (MPO), proteinase 3 (PR3), bactericidal/permeability-increasing protein (BPI), lactoferrin (LF), leukocyte elastase (LE), and cathepsin G (CG). The IgG subclass reactivity of antibodies to BPI and LF was measured. RESULTS: Antineutrophil cytoplasmic antibodies were found in 18 RA patients. Only a P-ANCA fluorescence pattern was seen. Six sera reacted to BPI, five to LF, one to MPO, one to PR3, and one to CG by ELISA testing. Immunoglobulin-G1 was the predominant subclass for LF-ANCA, whereas IgG1/3 contributed mainly to BPI-ANCA. Compared to P-ANCA-negative RA patients, the P-ANCA-positive patients exhibited significantly higher inflammatory activity, as estimated by disease activity score, C-reactive protein, erythrocyte sedimentation rate, and higher levels of IgM rheumatoid factor. CONCLUSION: Twenty percent of Bulgarian patients with RA have P-ANCA in their sera. These antibodies are directed against variable antigen specificities, while ANCA positivity in RA reflects disease and inflammatory activity. PMID- 16341905 TI - 4-methylumbelliferone, a hyaluronan synthase suppressor, enhances the anticancer activity of gemcitabine in human pancreatic cancer cells. AB - Hyaluronan (HA) is a ubiquitous, major component of the pericellular matrix and is necessary for various physiological processes. It plays a very important role in biological barriers. We previously reported that 4-methylumbelliferone (MU) inhibits HA synthesis and pericellular HA matrix formation in cultured human skin fibroblasts, Streptococcus equi FM100, and B16F10 melanoma cells. We hypothesized that MU-mediated inhibition of HA synthesis and pericellular HA matrix formation would increase the efficacy of anticancer drugs. We have already demonstrated in vitro, using a sandwich binding protein assay and a particle exclusion assay, that MU inhibits HA synthesis and formation of the pericellular HA matrix, respectively, in human KP1-NL pancreatic cancer cells. AlamarBlue assay revealed that the anticancer effect of gemcitabine in KP1-NL cells was increased by pretreatment with MU. In vivo simultaneous administration of MU and gemcitabine to tumor-bearing mice with severe combined immunodeficiency disease (SCID) decreased the size of the primary and metastatic tumors more than did gemcitabine alone. These data strongly suggest that a combination of MU and gemcitabine is effective against human pancreatic cancer cells. MU may have potential as a chemosensitizer and may provide us with a new anticancer strategy. PMID- 16341906 TI - Introduction to the special issue: human linkage studies for behavioral traits. AB - In the post Genome era, the aim of behavior genetics has shifted from estimating the relative contributions of genes and environmental factors to (co-)variation in human complex traits, to localization of genes and identification of functional genetic variants. This special issue reflects this transition and presents fifteen papers that report on genome-wide linkage scans for complex traits in humans and on methodological tools and innovations. Six papers focus on cognition and report overlapping linkage peaks on chromosomes 6p and 14p. Papers on addictive behavior, i.e. smoking and alcohol dependence and its endophenotypes, find moderate LOD scores on chromosomes 6p, 5q, 4p and 7q, respectively. Three papers concentrate on emotionality, depression and loneliness and examine chromosomes 2q and 12q. The papers in this issue represent a summary of the first large scale linkage enterprises of human behavioral traits. PMID- 16341907 TI - Linkage analyses of IQ in the collaborative study on the genetics of alcoholism (COGA) sample. AB - Intelligence, as measured by standardized psychological tests, has been shown to be highly heritable, though identifying specific genes influencing general intelligence has proven difficult. We conducted genome-wide linkage analyses to identify chromosomal regions containing genes influencing intelligence, as measured by WAIS full-scale IQ (FSIQ), performance IQ (PIQ) and verbal IQ (VIQ). Non-parametric multipoint linkage analyses were conducted with Merlin-regress software, using a sample of 1,111 genotyped and phenotyped individuals from 201 families, ascertained as part of the Collaborative Study on the Genetics of Alcoholism (COGA). The strongest evidence of linkage was obtained for FSIQ on chromosome 6 (LOD=3.28, 12 cM) near the marker D6S1006. This region was also implicated with suggestive linkage in a recently published genome screen of IQ in Australian and Dutch twin pairs, and it has been implicated in linkage studies of developmental dyslexia. Our findings provide further support that chromosome 6p contains gene(s) affecting intelligence. PMID- 16341908 TI - Cognitive traits link to human chromosomal regions. AB - : Human cognition in normal and disease states is both environmentally and genetically mediated. Except for measures of language-specific abilities, however, few cognitive measures have been associated with specific genes or chromosomal regions. We performed genome-wide linkage analysis of five neuropsychological tests in the Collaborative Study on Genetics of Alcoholism sample. The sample included 1579 individuals (53% female, 76% White Non-Hispanic) in 217 families. There were 390 markers with mean inter-marker distance of 9.6 cM. Performance on the Digit Symbol Substitution Test, a component of the Wechsler Adult Intelligence Scale-R, showed significant linkage to 14q11.2 and suggestive linkage to 14q 24.2. This test of sustained visual attention also involves visual-motor coordination and executive functions. Performance on the WAIS-R Digit Span Test of immediate memory and mental flexibility showed suggestive linkage to 11q 25. Although the validity of these results beyond populations with a susceptibility for alcohol dependence is unclear, these results are among the first linkage results for non-language components of cognition. PMID- 16341909 TI - Endophenotypes successfully lead to gene identification: results from the collaborative study on the genetics of alcoholism. AB - The use of endophenotypes has been proposed as a strategy to aid gene identification efforts for complex phenotypes [Gottesman, I. I., and Shields J. (1972). Schizophrenia and Genetics: A Twin Study Vantage Point. London: Academic]. As part of the Collaborative Study of the Genetics of Alcoholism (COGA) project, we have analyzed electrophysiological endophenotypes, in addition to clinical diagnoses, as part of our effort to identify genes involved in the predisposition to alcohol dependence. In this paper we summarize published results from linkage and association analyses of two chromosomal regions in which the use of endophenotypes has successfully led to the identification of genes associated with alcohol dependence [GABRA2 (Edenberg et al., (2004). Am. J. Hum. Genet. 74:705-714) and CHRM2 (Wang et al., (2004). Hum. Mol. Genet. 13:1903 1911)]. Our experience in the COGA project has been that the analysis of endophenotypes provides several advantages over diagnostic phenotypes, including the strength and localization of the linkage signal. Our results provide an illustration of the successful use of endophenotypes to identify genes involved in the predisposition to a complex psychiatric phenotype, a strategy originally proposed by Gottesman and Shields in 1972. PMID- 16341910 TI - Correlates of early postpartum depressive symptoms. AB - BACKGROUND: Postpartum depressive symptoms negatively affect the quality of life and daily functioning of mothers and infants. Little research has examined the impact of situational factors such as physical symptom burden and function on early postpartum depressive symptoms. OBJECTIVES: To explore the association between situational factors and early postpartum depressive symptoms. To predict correlates of early postpartum depressive symptoms. METHODS: Cross sectional telephone survey of 720 mothers between 2 and 6 weeks postpartum. Mothers reported on demographic factors, physical and psychological symptoms, daily function, infant behaviors, social support, and skills in managing infant and household. The association between these factors and postpartum depressive symptoms were investigated with bivariate and multivariable analyses. RESULTS: Nearly, 39% of patients screened positive for depressive symptoms. Bivariate analyses showed symptomatic patients were more likely to be nonwhite, have lower incomes, less education, a past history of depression, have higher physical symptom burdens, more physical functional limitations, more infant colic, receive less social support, and have lower-self-efficacy scores as compared with patients without symptoms of depression. In a multivariable model predicting depressive symptoms, nonwhite race (odds ratio (OR) of 1.96, 95% confidence interval (CI) of 1.38, 2.78), more physical symptoms (OR 1.16, 95% CI 1.01, 1.32), infant colic (OR 1.79, 95% CI 1.29, 2.50) lack of social support (OR 0.90, 95% CI 0.86, 0.95), and lower self-efficacy scores (OR 0.90, 95% CI 0.86, 0.94) were associated with depressive symptoms. CONCLUSIONS: Nonwhite race, physical symptom burden, infant colic, lack of social support, and lower self-efficacy scores are associated with early postpartum depressive symptoms. Further research is needed to investigate whether providing social support and teaching skills to enhance self-efficacy will reduce the incidence of early postpartum symptoms of depression. PMID- 16341911 TI - Adapting smoking relapse-prevention materials for pregnant and postpartum women: formative research. AB - OBJECTIVE: To decrease smoking relapse among pregnant and postpartum women by adapting existing, validated relapse-prevention materials to meet the unique needs of pregnant and postpartum women. METHODS: A series of semi-structured interviews and learner verification activities were conducted with pregnant abstinent, postpartum abstinent, and postpartum relapsed women. Results were used to create new relapse-prevention materials, specific to the needs of pregnant and postpartum women, which are currently being used in a randomized clinical trial. RESULTS: Findings are consistent with the recurrent themes in the literature regarding smoking cessation among pregnant and postpartum women and revealed exceptional needs for coping and stress reduction strategies related to remaining abstinent postpartum. Conflict levels were also high in areas of identity, social support, and reasons for quitting. CONCLUSION: By interviewing women about their cessation related needs, the current study was able to produce smoking relapse prevention materials specific to this population. Having pregnant and postpartum women review the modified program materials before starting the clinical trial enhanced the quality, dependability, and validity of the materials. We await the results of the clinical trial to determine if this intervention is indeed more efficacious than previous attempts to intervene with this population. PMID- 16341912 TI - Conceptual versus perceptual priming in incomplete picture identification. AB - This paper examined conceptual versus perceptual priming in identification of incomplete pictures by using a short-term priming paradigm, in which information that may be useful in identifying a fragmented target is presented just prior to the target's presentation. The target was a picture that slowly and continuously became complete and the participants were required to press a key as soon as they knew what it was. Each target was preceded by a visual prime. The nature of this prime varied from very conceptual (e.g., the name of the picture's category) to very perceptual (e.g., a similar-shaped pictorial prime from a different category). Primes also included those that combined perceptual and conceptual information (e.g., names or images of the target picture). Across three experiments, conceptual primes were effective while the purely perceptual primes were not. Accordingly, we conclude that pictures in this type of task are identified primarily by conceptual processing, with perceptual processing contributing relatively little. PMID- 16341913 TI - The effects of articulatory suppression on word recognition in Serbian. AB - The relatedness of phonological coding to the articulatory mechanisms in visual word recognition vary in different writing systems. While articulatory suppression (i.e., continuous verbalising during a visual word processing task) has a detrimental effect on the processing of Japanese words printed in regular syllabic Khana script, it has no such effect on the processing of irregular alphabetic English words. Besner (1990) proposed an experiment in the Serbian language, written in Cyrillic and Roman regular but alphabetic scripts, to disentangle the importance of script regularity vs. the syllabic-alphabetic dimension for the effects observed. Articulatory suppression had an equally detrimental effect in a lexical decision task for both alphabetically regular and distorted (by a mixture of the two alphabets) Serbian words, but comparisons of articulatory suppression effect size obtained in Serbian to those obtained in English and Japanese suggest "alphabeticity-syllabicity" to be the more critical dimension in determining the relatedness of phonological coding and articulatory activity. PMID- 16341914 TI - Uh and um revisited: are they interjections for signaling delay? AB - Clark and Fox Tree (2002) have presented empirical evidence, based primarily on the London-Lund corpus (LL; Svartvik & Quirk, 1980), that the fillers uh and um are conventional English words that signal a speaker's intention to initiate a minor and a major delay, respectively. We present here empirical analyses of uh and um and of silent pauses (delays) immediately following them in six media interviews of Hillary Clinton. Our evidence indicates that uh and um cannot serve as signals of upcoming delay, let alone signal it differentially: In most cases, both uh and um were not followed by a silent pause, that is, there was no delay at all; the silent pauses that did occur after um were too short to be counted as major delays; finally, the distributions of durations of silent pauses after uh and um were almost entirely overlapping and could therefore not have served as reliable predictors for a listener. The discrepancies between Clark and Fox Tree's findings and ours are largely a consequence of the fact that their LL analyses reflect the perceptions of professional coders, whereas our data were analyzed by means of acoustic measurements with the PRAAT software (www.praat.org). A comparison of our findings with those of O'Connell, Kowal, and Ageneau (2005) did not corroborate the hypothesis of Clark and Fox Tree that uh and um are interjections: Fillers occurred typically in initial, interjections in medial positions; fillers did not constitute an integral turn by themselves, whereas interjections did; fillers never initiated cited speech, whereas interjections did; and fillers did not signal emotion, whereas interjections did. Clark and Fox Tree's analyses were embedded within a theory of ideal delivery that we find inappropriate for the explication of these phenomena. PMID- 16341915 TI - The dependency structure of coordinate phrases: a corpus approach. AB - Hudson (1990) proposes that each conjunct in a coordinate phrase forms dependency relations with heads or dependents outside the coordinate phrase (the "multi head" view). This proposal is tested through corpus analysis of Wall Street Journal text. For right-branching constituents (such as direct-object NPs), a short-long preference for conjunct ordering is observed; this is predicted by the multi-head view, under the assumption that structures resulting in shorter dependencies are preferred. A short-long preference is also observed for left branching constituents (such as subject NPs), which is less obviously accommodated by the multi-head view but not incompatible with it. The repetition of determiners was also examined (the dog and cat versus the dog and the cat), and a stronger preference was found for repetition with singular count nouns as opposed to mass or plural nouns; this accords well with the multi-head view, under the reasoning that single-determiner constructions require crossing dependencies with count nouns but not with plural or mass nouns. PMID- 16341916 TI - Do Chinese dyslexic children have difficulties learning English as a second language? AB - The aim of the present study was to examine whether Chinese dyslexic children had difficulties learning English as a second language given the distinctive characteristics of the two scripts. Twenty-five Chinese primary school children with developmental dyslexia and 25 normally achieving children were tested on a number of English vocabulary, reading, and phonological processing tasks. It was found that the Dyslexia group performed significantly worse than the Control group in nearly all the English measures. The findings suggest that Chinese dyslexic children also encounter difficulties in learning English as a second language, and they are generally weak in phonological processing both in Chinese and English. However, phonological skills were found to correlate significantly with English reading but not with Chinese reading in the dyslexic children. It is evident that there are both common and specific causes to reading difficulties in Chinese and English. PMID- 16341917 TI - Mechanobiology in the third dimension. AB - Cells are mechanically coupled to their extracellular environments, which play critical roles in both communicating the state of the mechanical environment to the cell as well as in mediating cellular response to a variety of stimuli. Along with the molecular composition and mechanical properties of the extracellular matrix (ECM), recent work has demonstrated the importance of dimensionality in cell-ECM associations for controlling the sensitive communication between cells and the ECM. Matrix forces are generally transmitted to cells differently when the cells are on two-dimensional (2D) vs. within three-dimensional (3D) matrices, and cells in 3D environments may experience mechanical signaling that is unique vis-a-vis cells in 2D environments, such as the recently described 3D-matrix adhesion assemblies. This review examines how the dimensionality of the extracellular environment can affect in vitro cell mechanobiology, focusing on collagen and fibrin systems. PMID- 16341918 TI - Macromolecular transport in the arterial wall: alternative models for estimating barriers. AB - Early atherosclerosis, or atherogenesis, is characterized by the abnormal accumulation of plasma-borne macromolecules (e.g., LDL) in the arterial intima. The change of barrier characteristics of tissue in the arterial wall requires evaluation of macromolecular transport across the endothelial cell layer (ECL) and internal elastic lamina (IEL), the luminal and abluminal boundaries of the arterial intima, respectively. In this study, alternative mathematical models are derived from dynamic mass balances to describe macromolecular transport across the arterial wall. One model considers each medial layer as a spatially lumped compartment, whereas another model consists of a spatially lumped intima and spatially distributed media. Model simulations of a tracer concentration distribution in the arterial wall are compared with concentration distributions of horseradish peroxidase (HRP) after i.v. injection in mice. For each model, optimal parameter values are obtained that yield model outputs matching the data well for two different HRP circulation times. The model parameter estimates show that the ECL is the major barrier for macromolecular transport across the normal arterial wall. Sensitivity analysis indicates that the parameter estimates of the transport coefficients of the ECL and IEL are well determined. Optimal circulation times are determined and expected to yield improved precision of parameter estimates in future experiments to reflect disease progression. PMID- 16341919 TI - Post-vagotomy mechanical characteristics and structure of the thoracic aortic wall. AB - This study assessed the long-term effect of vagotomy on the structure and passive mechanical characteristics of the thoracic aorta under a wide range of stresses in vitro. Eight healthy Landrace pigs underwent bilateral vagotomy distal to the origin of the recurrent laryngeal nerve, and 10 pigs were sham-operated. Three months post-surgery, the aorta was excised and specimens from the ascending aorta, arch, and descending thoracic aorta were subjected to histomorphometrical evaluation and uniaxial tensile-testing until failure. Elastic modulus-stress data were plotted and submitted to regression analysis. Structural remodeling after vagotomy was characterized as vascular growth in the ascending aorta and arch, and as thinning in the descending thoracic aorta. In the aortic segments of vagotomized animals, the area density of elastin and collagen was increased, but smooth muscle density was decreased. Similar differences in regression parameters and failure strength between groups were found in all aortic segments, indicating that the vessel wall was stiffer and stronger in vagotomized animals. In the clinical setting, disease states or drugs blocking the regulatory role of the vagi nerves on the aortic wall may have undesirable consequences on the mechanical performance of the thoracic aorta, and therefore on hemodynamic homeostasis. PMID- 16341920 TI - Large-scale 3-D geometric reconstruction of the porcine coronary arterial vasculature based on detailed anatomical data. AB - The temporal and spatial distribution of coronary blood flow, pressure, and volume are determined by the branching pattern and three-dimensional (3-D) geometry of the coronary vasculature, and by the mechanics of heart wall and vascular tone. Consequently, a realistic simulation of coronary blood flow requires, as a first step, an accurate representation of the coronary vasculature in a 3-D model of the beating heart. In the present study, a large-scale stochastic reconstruction of the asymmetric coronary arterial trees (right coronary artery, RCA; left anterior descending, LAD; and left circumflex, LCx) of the porcine heart has been carried out to set the stage for future hemodynamic analysis. The model spans the entire coronary arterial tree down to the capillary vessels. The 3-D tree structure was reconstructed initially in rectangular slab geometry by means of global geometrical optimization using parallel simulated annealing (SA) algorithm. The SA optimization was subject to constraints prescribed by previously measured morphometric features of the coronary arterial trees. Subsequently, the reconstructed trees were mapped onto a prolate spheroid geometry of the heart. The transformed geometry was determined through least squares minimization of the related changes in both segments lengths and their angular characteristics. Vessel diameters were assigned based on a novel representation of diameter asymmetry along bifurcations. The reconstructed RCA, LAD and LCx arterial trees show qualitative resemblance to native coronary networks, and their morphological statistics are consistent with the measured data. The present model constitutes the first most extensive reconstruction of the entire coronary arterial system which will serve as a geometric foundation for future studies of flow in an anatomically accurate 3-D coronary vascular model. PMID- 16341921 TI - Shear stress regulates HUVEC hydraulic conductivity by occludin phosphorylation. AB - Human umbilical vein endothelial cells (HUVECs) display hydraulic conductivity (L(P)) responses to shear stress that differ markedly from the responses of bovine aortic endothelial cells (BAECs). In HUVECs, 5, 10, and 20 dyn cm(-2) steady shear stress transiently increased L(P) with a return to preshear baseline after a 2-h exposure to shear stress. Pure oscillatory shear stress of 0 +/- 20 dyn cm(-2) (mean+/-amplitude) had no effect on L(P), whereas superposition of oscillatory shear stress on steady shear stress suppressed the effect induced by steady shear stress alone. Shear reversal (amplitude greater than mean) was not necessary for the inhibitory influence of oscillatory shear stress. The transient increase of L(P) by steady shear stress was not affected by incubation with BAPTA AM (10 microM), suggesting calcium independence of the shear response. Decreasing nitric oxide (NO) concentration with L-NMMA (100 microM), a nitric oxide synthase (NOS) inhibitor, did not inhibit the HUVEC L(P) response to shear stress. At the protein level, 10 dyn cm(-2) shear stress did not affect the total content of occludin, but it did elevate the phosphorylation level transiently. The positive correlation between occludin phosphorylation and hydraulic conductivity parallels observations in BAECs and suggests that occludin phosphorylation may be a general mediator of shear-L(P) responses in diverse endothelial cell types. PMID- 16341922 TI - Oxidative stress produced with cell migration increases synthetic phenotype of vascular smooth muscle cells. AB - Phenotypic modulation of vascular smooth muscle cells (VSMC) and reactive oxygen species (ROS) is important in vascular pathogenesis. Understanding how these factors relate to cell migration can improve design of therapeutic interventions to control vascular disease. We compared the proliferation, protein content and migration of cultured aortic VSMC from wild type (WT) versus transgenic mice (Tgp22phox), in which overexpression of p22phox was targeted to VSMC. Also, we compared H2O2 generation and expression of specific phenotypic markers of non migrating with migrating WT versus Tgp22phox VSMC in an in vitro wound scratch model. Enhanced H2O2 production in Tgp22phox versus WT VSMC (p < 0.005) significantly correlated with increased protein content, proliferation, and migration. VSMC migrating across the wound edge produced more H2O2 than non migrating VSMC (p < 0.05). The expression of synthetic phenotypic markers, tropomyosin 4 and myosin heavy chain embryonic (SMemb), was enhanced significantly, while the expression of contractile marker, smooth muscle alpha actin, was reduced significantly in migrating versus non-migrating cells, and also in Tgp22phox versus WT (p < 0.005) VSMC. These results are consistent with increased production of ROS accelerating the switch from the contractile to the synthetic phenotype, characterized by increases in proliferation, migration, and expression of TM4 and SMemb and decreased alpha-actin. PMID- 16341923 TI - A LabVIEW model incorporating an open-loop arterial impedance and a closed-loop circulatory system. AB - While numerous computer models exist for the circulatory system, many are limited in scope, contain unwanted features or incorporate complex components specific to unique experimental situations. Our purpose was to develop a basic, yet multifaceted, computer model of the left heart and systemic circulation in LabVIEW having universal appeal without sacrificing crucial physiologic features. The program we developed employs Windkessel-type impedance models in several open loop configurations and a closed-loop model coupling a lumped impedance and ventricular pressure source. The open-loop impedance models demonstrate afterload effects on arbitrary aortic pressure/flow inputs. The closed-loop model catalogs the major circulatory waveforms with changes in afterload, preload, and left heart properties. Our model provides an avenue for expanding the use of the ventricular equations through closed-loop coupling that includes a basic coronary circuit. Tested values used for the afterload components and the effects of afterload parameter changes on various waveforms are consistent with published data. We conclude that this model offers the ability to alter several circulatory factors and digitally catalog the most salient features of the pressure/flow waveforms employing a user-friendly platform. These features make the model a useful instructional tool for students as well as a simple experimental tool for cardiovascular research. PMID- 16341924 TI - Lyapunov exponents of laser Doppler flowmetry signals in healthy and type 1 diabetic subjects. AB - The skin of diabetic subjects presents abnormalities in capillary blood flow and its regulation, often leading to the generation of plantar ulcers. In order to gain insight into this pathology for type 1 diabetic patients, Lyapunov exponents (LEs) of signals reflecting microvascular perfusion--laser Doppler flowmetry (LDF) signals--are calculated. The algorithm to compute LEs is first validated on simulated data and LDF surrogates. Then, LDF signals recorded at rest and during the application of local and progressive pressure of 11.1 Pa/s are processed. The exponents appear in pairs and are different for healthy and type 1 diabetic subjects at rest; P = 0.0556 for the 7th, 8th, and 9th LEs. Furthermore, progressive pressure has also a distinct effect on LEs. The difference is more pronounced for diabetic patients, for whom P = 0.0625 for the four LEs of highest absolute value. Because these differences arise from abnormalities in microvascular blood flow, they may help to explain the high prevalence of type 1 diabetic patients developing foot ulcers. PMID- 16341925 TI - Estimation of time-varying coherence function using time-varying transfer functions. AB - We introduce a new method to estimate reliable time-varying coherence functions (TVCF) for causal systems. The technique is based on our previously developed method to estimate time-varying transfer functions (TVTF), known as the time varying optimal parameter search algorithm [Zou, R., H. Wang, and K. H. Chon. A robust time-varying identification algorithm using basis functions. Ann. Biomed. Eng. 31: 840-853, 2003]. The TVCF is estimated by the multiplication of two TVTFs. The two TVTFs are obtained using signal x as the input and signal y as the output to produce the first TVTF, and signal y as the input and signal x as the output to produce the second TVTF. Demonstration of the feasibility and efficacy of the proposed approach is provided with both simulation examples and application to renal blood flow and pressure data. The proposed approach provides higher time-frequency resolution TVCF than afforded by the short time Fourier transform based TVCF. PMID- 16341926 TI - Improving hollow fiber dialyzer efficiency with a recirculating dialysate system II: comparison against two-chamber dialysis systems. AB - The theoretical basis of the nonregenerated recirculating dialysate system (RDS) was derived in Part I of this work [M. Prado, L. M. Roa, A. Palma, and J. A. Milan, Ann. Biomed. Eng. (2005)]. This system pursues the maximization of the clearance of hollow fiber dialyzers whose performance is controlled by diffusion, as occurred in standard hemodialysis. In this second part we perform a comparison by digital simulation of the RDS against three well-known two-chamber dialysis systems. As a major outcome, the efficiency of the RDS increased by a factor of five-eight with respect to the efficiency of a single dialyzer operating with a number of transfer units equal to 0.1, that is when the diffusive mass-transfer of the dialyzer is exhausted. Present low-flux dialyzers do not take advantage of the full potential of this technique, but the functional domain where high-flux and high-area dialyzers operate could be more suitable to exploit this technique. We conclude that RDS can be a competitive efficient technique for optimizing the dialysis efficiency. PMID- 16341927 TI - Long-term mathematical model involving renal sympathetic nerve activity, arterial pressure, and sodium excretion. AB - This paper presents a physiological long-term model of the cardiovascular system. It integrates the previous models developed by Guyton, Uttamsingh and Coleman. Additionally it introduces mechanisms of direct effects of the renal sympathetic nerve activity (rsna) on tubular sodium reabsorption and renin secretion in accordance with experimental data from literature. The resulting mathematical model constitutes the first long-term model of the cardiovascular system accounting for the effects of rsna on kidney functions in such detail. The objective of developing such a model is to observe the consequences of long-term rsna increase and impairment of rsna inhibition under volume loading. This model provides an understanding of the rsna-related mechanisms, which cause mean arterial pressure increase in hypertension and total sodium amount increase (sodium retention) in congestive heart failure, nephrotic syndrome and cirrhosis. PMID- 16341928 TI - Tissue elasticity estimation with optical coherence elastography: toward mechanical characterization of in vivo soft tissue. AB - High-resolution imaging provides a significant means for accurate material modulus estimation and mechanical characterization. Within the realm of in vivo soft tissue characterization, particularly on small biological length scales such as arterial atherosclerotic plaques, optical coherence tomography (OCT) offers a desirable imaging modality with higher spatial resolution and contrast of tissue as compared with intravascular ultrasound (IVUS). Based on recent advances in OCT imaging and elastography, we present a fully integrated system for tissue elasticity reconstruction, and assess the benefits of OCT on the distribution results of four representative tissue block models. We demonstrate accuracy, with displacement residuals on the order of 10(-6) mm (more than 3 orders of magnitude less than average calculated displacements), and high-resolution estimates, with the ability to resolve inclusions of 0.15 mm diameter. PMID- 16341929 TI - A predictive model of therapeutic monoclonal antibody dynamics and regulation by the neonatal Fc receptor (FcRn). AB - We constructed a novel physiologically-based pharmacokinetic (PBPK) model for predicting interactions between the neonatal Fc receptor (FcRn) and anti carcinoembryonic antigen (CEA) monoclonal antibodies (mAbs) with varying affinity for FcRn. Our new model, an integration and extension of several previously published models, includes aspects of mAb-FcRn dynamics within intracellular compartments not represented in previous PBPK models. We added mechanistic structure that details internalization of class G immunoglobulins by endothelial cells, subsequent FcRn binding, recycling into plasma of FcRn-bound IgG and degradation of free endosomal IgG. Degradation in liver is explicitly represented along with the FcRn submodel in skin and muscle. A variable tumor mass submodel is also included, used to estimate the growth of an avascular, necrotic tumor core, providing a more realistic picture of mAb uptake by tumor. We fitted the new multiscale model to published anti-CEA mAb biodistribution data, i.e. concentration-time profiles in tumor and various healthy tissues in mice, providing new estimates of mAb-FcRn related kinetic parameters. The model was further validated by successful prediction of F(ab')2 mAb fragment biodistribution, providing additional evidence of its potential value in optimizing intact mAb and mAb fragment dosing for clinical imaging and immunotherapy applications. PMID- 16341930 TI - A general framework for characterizing studies of brain interface technology. AB - The development of brain interface (BI) technology continues to attract researchers with a wide range of backgrounds and expertise. Though the BI community is committed to accurate and objective evaluation of methods, systems, and technology, the very diversity of the methods and terminology used in the field hinders understanding and impairs technology cross-fertilization and cross group validation of findings. Underlying this dilemma is a lack of common perspective and language. As seen in our previous works in this area, our approach to remedy this problem is to propose language in the form of taxonomy and functional models. Our intent is to document and validate our best thinking in this area and publish a perspective that will stimulate discussion. We encourage others to do the same with the belief that focused discussion on language issues will accelerate the inherently slow natural evolution of language selection and thus alleviate related problems. In this work, we propose a theoretical framework for describing BI-technology-related studies. The proposed framework is based on the theoretical concepts and terminology from classical science, assistive technology development, human-computer interaction, and previous BI-related works. Using a representative set of studies from the literature, the proposed BI study framework was shown to be complete and appropriate perspective for thoroughly characterizing a BI study. We have also demonstrated that this BI study framework is useful for (1) objectively reviewing existing BI study designs and results, (2) comparing designs and results of multiple BI studies, (3) designing new studies or objectively reporting BI study results, and (4) facilitating intra- and inter-group communication and the education of new researchers. As such, it forms a sound and appropriate basis for community discussion. PMID- 16341931 TI - Expression of MARCKS effector domain mutants alters phospholipase D activity and cytoskeletal morphology of SK-N-MC neuroblastoma cells. AB - Stable overexpression of myristoylated alanine-rich C-kinase substrate (MARCKS) is known to enhance phorbol ester stimulation of phospholipase D (PLD) activity and protein kinase Calpha (PKCalpha) levels in SK-N-MC neuroblastoma cells. In contrast, expression of MARCKS mutants (S152A or S156A) lacking key PKC phosphorylation sites within the central basic effector domain (ED) had no significant effect on PLD activity or PKCalpha levels relative to vector control cells. Like control cells, those expressing wild type MARCKS were elongated and possessed longitudinally oriented stress fibers, although these cells were more prone to detach from the substratum and undergo cell death upon phorbol ester treatment. However, cells expressing MARCKS ED mutants were irregularly shaped and stress fibers were either shorter or less abundant, and cell adhesion and viability were not affected. These results suggest that intact phosphorylation sites within the MARCKS ED are required for PLD activation and influence both membrane-cytoskeletal organization and cell viability. PMID- 16341932 TI - Is the platelet serotonin transporter different in venous vs. arterial blood? AB - The binding of labelled paroxetine to the serotonin transporter (SERT) of platelet membranes has been studied in both venous and mixed venous/arterial blood of the rat. In addition, we studied the inhibition of paroxetine binding to SERT by quipazine and N-methyl-quipazine (NMQ). The results indicate differences in affinity for the two test drugs, quipazine and NMQ, in venous vs. mixed venous/arterial blood. This suggests different post-translational modifications of SERT in platelets of arterial vs. venous blood. PMID- 16341933 TI - PPAR-alpha expression inversely correlates with inflammatory cytokines IL-1beta and TNF-alpha in aging rats. AB - Dehydroepiandrosterone (DHEAS) was given the name "fountain of youth" in reference to its beneficial properties in memory, cognition and aging. Cultured cell studies showed that DHEAS may mediate its action by counteracting aging associated inflammation via PPAR-alpha activation. In the present study, we demonstrated an age-dependent increase in IL-1beta and TNF-alpha expression in the brain and the spleen of aging rats, while PPAR-alpha expression was decreased in the spleen of 18 month-old rats. Oral treatment with DHEAS increased PPAR alpha mRNA in 3 month-old rats and decreased PPAR-alpha protein expression in 18 month-old rats in the spleen. In contrast, DHEAS did not alter cytokine expression in spleen and brain of the three age groups. These findings underline a differential role for DHEAS in PPAR-alpha expression that is age-dependent, and also, that beneficial effects of DHEAS on cognitive function are unlikely mediated by a decrease in cytokine expression. PMID- 16341934 TI - Effects of monoamine oxidase inhibitors on methamphetamine-induced stereotypy in mice and rats. AB - In male ICR mice, a single intraperitoneal administration of methamphetamine (METH) (10 mg/kg) induced stereotyped behavior such as continuous sniffing, circling, and nail biting, reaching a plateau level 20 min after the injection. Subcutaneous pretreatment with clorgyline, a monoamine oxidase (MAO)-A inhibitor, at a dose of 0.1 mg/kg 2 h prior to the drug challenge significantly decreased the initial (first 20 min) intensity of stereotypies and increased the latency to onset. The effect was not observed with either higher doses of clorgyline (1 and 10 mg/kg) or l-deprenyl, a MAO-B inhibitor, at doses of 0.1-10 mg/kg. In male Wistar rats, the inhibitory effect of clorgyline on METH-induced stereotypy was not observed. Pretreatment of the mice with clorgyline (0.1 mg/kg) had no effect on apparent serotonin and dopamine turnover in the striatum, although the higher doses of clorgyline (1 and 10 mg/kg) significantly decreased the turnover. These results suggest that a low dose of clorgyline tends to increase the latency and decrease the intensity of stereotypies induced by METH in a dopamine metabolism independent manner in mice. PMID- 16341935 TI - Allyl chloride-induced time dependent changes of lipid peroxidation in rat nerve tissue. AB - To accurately know the time-dependent changes of the lipid peroxidation and antioxidative status for elucidating the mechanism of neuropathy induced by allyl chloride (AC), the malondialdehyde (MDA), anti-reactive oxygen species (anti ROS), glutathione (GSH), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) were investigated in cerebrum, spinal cord and sciatic nerve of rats after 0, 3, 6, 9, 12 weeks of AC administration. AC was administrated to Wistar rats by gavage at a single dosage of 200 mg/kg/per dose (three times per week). Rats were sacrificed after 0, 3, 6, 9, 12 weeks of treatment, and cerebrum, spinal cord, sciatic nerves were dissected, homogenized and used for the determination of lipid peroxidation and antioxidative status. The results showed that MDA in cerebrum (112.4%) and sciatic nerve (113.1%) significantly increased (P<0.05) on third week of AC treatment and at gait score of 2, and further changes of MDA were observed after 6, 9, 12 weeks and at gait score of 3, 4. While a decrease (P<0.05) in the activities of GSH, CAT, GPx and SOD after 6, 9, 12 weeks intoxication and at gait score of 2, 3, 4 were observed in cerebrum, spinal cord and sciatic nerve. Anti-ROS activities also decreased in all three nerve tissues after 3, 6, 9, 12 weeks intoxication and at gait score of 2, 3, 4. Thus, AC intoxication was associated with elevation of lipid peroxidation and reduction of antioxidative status, and the time-dependent changes of these indexes in Wistar rats nerve tissues occurred. Sciatic nerve was the main target tissue and MDA was most sensitive among all indexes. The changes of lipid peroxidation and antioxidative status might be related to the degradation of nerve fiber and served as one of mechanisms of toxic neuropathy induced by AC. PMID- 16341936 TI - Evidence for a role of second pathophysiological stress in prevention of delayed neuronal death in the hippocampal CA1 region. AB - In ischemic tolerance experiment, when we applied 5-min ischemia 2 days before 30 min ischemia, we achieved a remarkable (95.8%) survival of CA1 neurons. However, when we applied 5-min ischemia itself, without following lethal ischemia, we found out 45.8% degeneration of neurons in the CA1. This means that salvage of 40% CA1 neurons from postischemic degeneration was initiated by the second pathophysiological stress. These findings encouraged us to hypothesize that the second pathophysiological stress used 48 h after lethal ischemia can be efficient in prevention of delayed neuronal death. Our results demonstrate that whereas 8 min of lethal ischemia destroys 49.9% of CAI neurons, 10 min of ischemia destroys 71.6% of CA1 neurons, three different techniques of the second pathophysiological stress are able to protect against both: CA1 damage as well as spatial learning/memory dysfunction. Bolus of norepinephrine (3.1 micromol/kg i.p.) used two days after 8 min ischemia saved 94.2%, 6 min ischemia applied 2 days after 10 min ischemia rescued 89.9%, and an injection of 3-nitropropionic acid (20 mg/kg i.p.) applied two days after 10 min ischemia protected 77.5% of CA1 neurons. Thus, the second pathophysiological stress, if applied at a suitable time after lethal ischemia, represents a significant therapeutic window to opportunity for salvaging neurons in the hippocampal CA1 region against delayed neuronal death. PMID- 16341937 TI - NMDA receptors mediate consolidation of contextual memory in the hippocampus after context preexposure. AB - Male Wistar rats received bilateral infusions of vehicle (VEH) or aminophosphonopentanoic acid (AP5), an N-metil-D-aspartate (NMDA) receptor antagonist, into the dorsal hippocampus immediately after inhibitory avoidance (IA) training. Intrahippocampal infusion of AP5 blocked 24 h IA retention. In the second experiment, animals were preexposed to the IA training context 24 h prior to training and received an infusion of either VEH or AP5 immediately after the preexposure trial and a second infusion of VEH or AP5 immediately after IA training. AP5 did not affect retention in animals preexposed to the IA box and given VEH after preexposure, but blocked retention when given after both preexposure and training. AP5 impaired retention in rats preexposed to an environment distinct from the IA box. These results suggest that NMDA receptors in the dorsal hippocampus mediate the formation of a contextual representation of the task environment. PMID- 16341938 TI - Tuberin--a new molecular target in Alzheimer's disease? AB - Tuberous sclerosis complex (TSC) is a common genetic disorder in which affected individuals develop mental retardation, developmental brain defects and seizures. The TSC gene products, hamartin and tuberin, form a complex, of which tuberin is assumed to be the functional component being involved in a wide variety of different cellular processes. Here we report that tuberin protein levels are decreased in the frontal cortex of patients with Alzheimer's disease. In addition, tuberin levels are also decreased in Down syndrome brain samples positive for beta-amyloid plaques and neurofibrillary tangles. Analysis of NeuN revealed that this regulation is not a consequence of differences in the amount of postmitotic neurons. This first connection of tuberin to another common disease beside TSC stimulates new approaches to investigate the molecular development and to establish new therapeutic strategies. PMID- 16341939 TI - Effects of mitochondrial toxins on the brain amino acid concentrations. AB - In the pathogenesis of Parkinson's disease and Huntington's disease excitotoxicity may play an important role. The common toxin model for Parkinson's disease is MPTP, while for Huntington's disease it is 3-NP. These toxins inhibit the mitochondrial respiratory chain, resulting in an energy deficit. In the central nervous system, the amino acids act as neurotransmitters and neuromodulators. The energy deficit caused by these neurotoxins may alter the concentrations of amino acids. Thus, it can be claimed that the aminoacidergic neurotransmission can be changed by neurotoxins. To test this hypothesis we studied the amino acid concentrations in different brain regions following MPTP or 3-NP administration. The two toxins were found to produce similar changes. We detected marked decreases in most of the amino acid concentrations in the striatum and in the cortex, while the levels in the cerebellum increased significantly. The decreased amino acid levels can be explained by the reduced levels of ATP produced by these neurotoxins. In the cerebellum, where there is no detectable ATP loss, the elevated amino acid levels may reflect a compensation of the altered neurotransmission. PMID- 16341940 TI - Impact of venlafaxine on gene expression profile in lymphocytes of the elderly with major depression--evolution of antidepressants and the role of the "neuro immune" system. AB - Antidepressive drugs offer considerable symptomatic relief in mood disorders and, although commonly discovered by screening with single biological targets, most interact with multiple receptors and signaling pathways. Antidepressants require a treatment regimen of several weeks before clinical efficacy is achieved in patient populations. While the biochemical mechanisms underlying the delayed temporal profile remain unclear, molecular adaptations over time are likely involved. The selective serotonin and noradrenaline reuptake inhibitor, venlafaxine, offers a dual antidepressive action. Its pharmacological behavior, however, is unknown at the genetic level, and it is difficult to monitor in human brain samples. Because the hypothalamic-pituitary-adrenal axis is often severely disrupted in mood disorders, lymphocytes may serve as models of neuropsychiatric conditions. As such, we examined the role of venlafaxine on the gene expression profile of human lymphocytes. DNA microarray was used to measure the expression patterns of multiple genes in human lymphocytes from depressed patients treated with this mood stabilizer. In this self-controlled study, RNAs of control and treated samples were purified, converted into cDNA and labeled with either Cy3 or Cy5, mixed and hybridized to DNA microarrays containing human oligonucleotides corresponding to more than 8,000 genes. Genes that were differentially regulated in response to treatment were selected for follow up on the basis on novelty, gene identity, and level of over-expression/repression, and selected transcripts were profiled by real-time PCR (data have been normalized to beta-actin). Using software analysis of the microarray data, a number of transcripts were differentially expressed between control and treated samples, of which only 57 were found to significantly vary with the "P" value of 0.05 or lower as a result of exposure to venlafaxine. Of these, 31 genes were more highly expressed and 26 transcripts were found to be significantly less abundant. Most selected genes were verified with QRT-PCR to alter. As such, independent verification using QRT PCR demonstrated the reliability of the method. Genes implicated in ionic homeostasis were differentially expressed, as were genes associated with cell survival, neural plasticity, signal transduction, and metabolism. Understanding how gene expression is altered over a clinically relevant time course of administration of venlafaxine may provide insight into the development of antidepressant efficacy as well as the underlying pathology of mood disorders. These changes in lymphocytes are thought to occur in the brain, and a "neuro immune system" is proposed by this study. PMID- 16341941 TI - In vitro, melatonin treatment decreases nitric oxide levels in murine splenocytes cultured with the venezuelan equine encephalomyelitis virus. AB - The purpose of this work was to determine the effect of melatonin on the nitric oxide levels in murine splenocytes cultured with the Venezuelan equine encephalomyelitis virus. After incubation, nitric oxide levels were measured by the diazotization assay. Those cultures with the Venezuelan equine encephalomyelitis virus increased nitric oxide levels. Splenocytes infected and treated with 100 and 150 microg/ml of melatonin, decreased significantly the nitric oxide levels when compared to infected and non-treated splenocytes. These findings show that splenocytes infected with the Venezuelan equine encephalomyelitis virus generate important amounts of nitric oxide and suggest that melatonin protects the mice infected with the Venezuelan equine encephalomyelitis virus by a mechanism involving the decreasing of nitric oxide concentrations in tissue. PMID- 16341942 TI - Glutamate metabolizing enzymes in prefrontal cortex of Alzheimer's disease patients. AB - Amounts of glutamate metabolizing enzymes such as glutamate dehydrogenase (GDH), glutamine synthetase (GS), GS-like protein (GSLP), and phosphate-activated glutaminase (PAG) were compared in prefrontal cortex of control subjects and patients with Alzheimer disease (AD). The target proteins were quantified by ECL Western immunoblotting in extracts from brain tissue prepared by two different techniques separating enzymes preferentially associated with cytoplasm (GDH I and II isoenzymes, GS, and partially GSLP) and membrane (GDH III, PAG, and partially GSLP) fractions. Amounts of all listed enzymes were found significantly increased in the patient group compared with controls. Some links between the measured values were observed in the control, but not in the AD patient group. The results may suggest for the pathological interruption of regulatory relations between distinct enzymes of glutamate metabolism in brain of AD patients. PMID- 16341943 TI - Brain oxygenation and metabolism during repetitive apnea with resuscitation of 21% and 100% oxygen in newborn piglets. AB - The oxygen distribution in the microcirculation of the piglet's brain and striatal extracellular dopamine were determined during repetitive apnea and resuscitation with 21% or 100% oxygen. Pre-apnea cortical oxygen was 49.5+/-10.4 mm Hg and during each apnea decreased to 8+/-0.9 mm Hg. After ten apneic episodes followed by resuscitation with 21% or 100% oxygen, 7.48+/-1.6% or 2.6+/-0.5% of the tissue volume was below 10 mm Hg, respectively. Extracellular dopamine increased progressively with an increase in the number of apneas with resuscitation of 21% oxygen and at the end of ten apneic episodes it was up to 59,500+/-11,320% of control. There was no increase in extracellular dopamine during apnea resuscitated with 100% oxygen. Repetitive apnea caused progressive increase in fraction of hypoxic brain tissue in newborn. The magnitude of the increase is dependent on whether the animals were resuscitated with room air or 100% oxygen. PMID- 16341945 TI - Our genes. PMID- 16341944 TI - The wide distribution of endornaviruses, large double-stranded RNA replicons with plasmid-like properties. AB - The International Committee on Taxonomy of Viruses (ICTV) recently accepted Endornavirus as a new genus of plant dsRNA virus. We have determined the partial nucleotide sequences of the RNA-dependent RNA polymerase regions from the large dsRNAs (about 14 kbp) isolated from barley (Hordeum vulgare), kidney bean (Phaseolus vulgaris), melon (Cucumis melo), bottle gourd (Lagenaria siceraria), Malabar spinach (Basella alba), seagrass (Zostera marina), and the fungus Helicobasidium mompa. Phylogenetic analyses of these seven dsRNAs indicate that these dsRNAs are new members of the genus Endornavirus that are widely distributed over the plant and fungal kingdoms. PMID- 16341946 TI - Gastroesophageal reflux disease: clinical features. AB - Gastroesophageal reflux disease (GERD) is a chronic disease affecting up to 40% of people in the Western world. Risk factors associated with GERD include age and lifestyle habits, although the clinically relevant contribution of many of these factors is unclear. In GERD, refluxed gastric acid damages the oesophageal mucosa, generally when the pH falls below 4. GERD patients present a variety of symptoms, most commonly heartburn and regurgitation. Oesophageal complications associated with GERD include erosions, ulcers, peptic strictures, and Barrett's oesophagus which is implicated in the development of oesophageal adenocarcinoma. Diagnosis of GERD is problematic due to the range of symptoms which may be presented to the physician and symptom severity is frequently unrelated to disease severity. While endoscopic monitoring may be used to assess the presence and severity of GERD, a lack of visible damage does not necessarily indicate an absence of GERD. Techniques used to diagnose GERD include addition of an acid solution into the oesophagus in order to replicate symptoms (Bernstein test) or 24-hour intra-oesophageal pH monitoring. Proton pump inhibitors are effective in the treatment of GERD, acting to reduce the acidity of the gastric juice and hence reduce oesophageal damage and symptoms associated with GERD. Symptoms most indicative of GERD are those associated with erosive oesophagitis, including heartburn and acid regurgitation. Less common GERD-associated symptoms include chest pain, a range of ear, nose and throat conditions, and asthma. In contrast to perceptions of the disease as 'merely' heartburn, the impact on patients' quality of life can be profound. Increasing awareness of GERD by health care professionals has led to improved diagnosis and a greater appreciation of the need for maintenance therapy. PMID- 16341947 TI - Clinical pharmacy at the university hospital from Sao Paulo, Brazil. AB - Clinical Pharmacy includes all services performed by pharmacists practicing in hospitals, community pharmacies, nursing homes, home-based care services, clinics and any other setting where medicines are prescribed and used. Clinical Pharmacy activities may influence the correct use of medicines at three different levels: before, during and after the prescription is written. At the same time, pharmaceutical care philosophy is coming to Brazil. This paper comments on the development of the Clinical Pharmacy concept in Brazil, mainly in its practice and experience in the University Hospital of Sao Paulo. PMID- 16341948 TI - Side effects of interferon-alpha therapy. AB - AIM: Interferon-alpha (IFN-alpha) has been extensively explored for its efficacy in various disease conditions and is currently used as a standard treatment in several of these. Its use is accompanied by a wide variety of possible side effects. These side-effects may hamper reaching and maintaining the dose needed for maximal therapeutic effect while their occurrence can outweigh clinical benefit of IFN-alpha treatment. This review addresses the toxicity profile of IFN alpha, the presumed pathophysiology of the different side effects and the strategies to handle these. METHODS: Computerized searches were used and cross references of articles and books were checked. RESULTS: Adverse effects due to IFN-alpha have been described in almost every organ system. Many side-effects are clearly dose-dependent. Taken together, occurrence of flu-like symptoms, hematological toxicity, elevated transaminases, nausea, fatigue, and psychiatric sequelae are the most frequently encountered. Although insight in the mechanisms accounting for IFN-alpha-related toxicities has improved in recent years, much remains to be elucidated. Guidelines on the management of these untoward sequelae are mostly based on clinical experience, while many side-effects can only be adequately handled by dose adjustment or cessation of treatment. CONCLUSION: Further research on the mechanisms underlying both therapeutic effects and adverse events is warranted. Hopefully, this will lead to better identification of those patients who are likely to benefit from treatment without experiencing severe toxicities. PMID- 16341949 TI - Treatment of gastroesophageal reflux disease. AB - AIM OF THE REVIEW AND METHODS: This review brings together information on the treatment of gastroesophageal reflux disease. Published manuscripts were identified from Medline. The articles were then screened for relevance prior to inclusion in the review. RESULTS AND CONCLUSION: Up to 40% of people in Western countries are estimated to regularly experience heartburn, the most characteristic symptom of gastroesophageal reflux disease (GERD). Treatment options available for GERD range from over-the-counter (OTC) antacids to proton pump inhibitors (PPIs) and anti-reflux surgery. Many patients self-medicate with OTC medications such as antacids and low-dose histamine H2-receptor antagonists (H2RA) to relieve episodic or food-related symptoms of GERD, and may not seek medical advice unless symptoms persist or worsen. However, GERD is a chronic disease that frequently affects health-related quality of life and, if not properly managed, the complications of GERD may include erosive oesophagitis (EO), Barrett's oesophagus and adenocarcinoma. Adequate control of acid secretion is key to the successful treatment of the condition. OTC medications provide effective symptom relief to about one quarter of patients suffering from GERD. H2RAs can also provide effective symptomatic relief, particularly in patients with milder GERD, but become less-effective over time. PPIs are the agents of choice for the suppression of gastric acid production and have become the mainstay of therapy for acid-related diseases. PPIs produce significantly faster and more complete symptomatic relief, significantly faster and more complete healing of erosive GERD compared with H2RAs and are also significantly more effective at preventing relapse of EO. There are a number of existing guidelines for the treatment of GERD. Recommendation for initial therapy consist of general measures, such as lifestyle advice in combination with antacids and/or alginates. When general measures fail, the next step is empirical therapy. Two options exist for the expirical treatment of GERD, so-called "step-up" or "step-down" therapy. There are no clear data to support either treatment strategy as a universal approach to all patients; consequently, physicians must always choose the course best suited to the individual patient. PMID- 16341950 TI - Survey of surgical antimicrobial prophylaxis in czech republic. AB - OBJECTIVE: To characterize the pattern of surgical antimicrobial prophylaxis in the Czech Republic. METHOD: Cross sectional survey with a self-administered postal questionnaire. Data collected included use of antimicrobial prophylaxis, surgical site infection rate, pathogens causing surgical site infection and demographics of the institution. Descriptive and multivariate analyses were performed. SETTING: Hospital, surgical departments in the Czech Republic. MAIN OUTCOME MEASURE: Prevalence of surgical antimicrobial use, factors associated with use, the profile of antimicrobial use, timing, route, dosage regimen and duration of initiated prophylaxis. RESULT: The response rate was 55.5%. Surgical antimicrobial prophylaxis was used in 97.5% of departments, and 85% departments justified prophylaxis based on guideline. The timing of the first dosage was within 2 h of operation in 95.0% of departments and 36.7% of all departments administered more than 2 doses of SAP in operations that lasted less than 4 h of all respondents. The three most common prophylactic antimicrobial agent used were cefazolin, co-amoxiclav and cefuroxime amongst the 26 single antimicrobial agents and 16 antimicrobial combinations. Penicillins and enzyme inhibitor was the most frequent class used. Surgical antimicrobial prophylaxis was administered intravenously in 82.5% of all cases. The regimen used varied markedly in dose and duration prescribed. The surgical site infection rate occurred. 1-5% in 71.7% of departments. Most departments identified the causative pathogen at all times. Staphylococcus aureus was the most frequent pathogen of surgical site infection and was detected in 90.8% of all departments. There was significant association between Pseudomonas aeruginosa with cefuroxime use and Bacteriodes fragilis with co amoxiclav use. CONCLUSION: This survey has identified several areas for improvement in surgical antimicrobial prophylaxis in the Czech Republic. Particular areas of concern include route of administration, duration and timing of first dosage of SAP, and the inappropriate use of broad-spectrum antimicrobials. PMID- 16341951 TI - Are essential medicines in Malaysia accessible, affordable and available? AB - OBJECTIVE: To assess the pharmaceutical sector to know whether people have access to essential medicines. SETTING: The study was conducted in 20 public health clinics, five public district drug stores and 20 private retail pharmacies selected randomly in five different areas randomly selected (four states and a federal territory). METHOD: The methodology used was adopted from the World Health Organization study protocol. The degree of attainment of the strategic pharmaceutical objectives of improved access is measured by a list of tested indicators. Access is measured in terms of the availability and affordability of essential medicines, especially to the poor and in the public sector. The first survey in the public health clinics and public district drug stores gathered information about current availability of essential medicines, prevalence of stock-outs and affordability of treatment (except drug stores). The second survey assessed affordability of treatment in public health clinics and private retail pharmacies. MAIN OUTCOME MEASURE: Availability, stock-out duration, percent of medicines dispensed, accessibility and affordability of key medicines. RESULTS: The average availability of key medicines in the public health clinics for the country was 95.4%. The average stock-out duration of key medicines was 6.5 days. However, average availability of key medicines in the public district drug stores was 89.2%; with an average stock-out duration of 32.4 days. Medicines prescribed were 100% dispensed to the patients. Average affordability for public health clinics was 1.5 weeks salary and for the private pharmacies, 3.7 weeks salary. CONCLUSIONS: The present pharmaceutical situation in the context of essential medicines list implementation reflected that the majority of the population in Malaysia had access to affordable essential medicines. If medicines need to be obtained from the private sector, they are hardly affordable. Although the average availability of essential medicines in Malaysia was high being more than 95.0%, in certain areas in Sabah availability was less than 80.0% and still a problem. PMID- 16341952 TI - Flemish community pharmacists' motivation and views related to continuing education. AB - OBJECTIVE: IPSA (Institute for Permanent Study for Pharmacists) wanted to assess community pharmacists' opinion on CE-related issues in order to develop more tailored CE programs. METHOD: A survey for self-administration was sent to 1691 community pharmacists. The method of stratified sampling was used to include pharmacists who do not take up CE courses as well. MAIN OUTCOME MEASURES: (1) Preferences for course formats, topics of interest and opinion on different CE providers; (2) Facilitators and barriers for participation in CE courses; (3) Opinion on rewarding and participation, obligation and willingness to pay. RESULTS: A response rate of 62.8% was obtained after three mailings. Lectures remain the most favorite course format. Topics related to pharmaceutical care are the most wanted. The strongest facilitators for attending CE courses are gathering practical knowledge and keeping scientific knowledge up to standard. Social contact with colleagues and receiving a syllabus are perceived as moderately motivating factors. Most frequently mentioned barriers were lack of time, family constraints, distance to the classes and uninteresting subjects. Around 2/3rd of questioned pharmacists are willing to invest more time in CE when participation would be rewarded. When it comes to opinion on mandatory CE, disagreement exists between pharmacists who take up CE courses and those who don't, the latter being less in favor. CONCLUSION: This survey revealed community pharmacists' opinions, wishes and needs concerning CE. This should enable CE providers to develop more tailored CE programs. PMID- 16341953 TI - A qualitative study of the contribution of pharmacists to heart failure management in Scotland. AB - STUDY OBJECTIVES: (1) To identify the medication management needs of chronic heart failure (CHF) patients and their caregivers; (2) To examine the perceived support for medication management available to these people from health professionals; (3) To identify the actual and potential perceived contribution of pharmacists to medication management. SETTING: A mixed urban/ rural region in the west of Scotland. DESIGN: Semi-structured qualitative research interviews. PARTICIPANTS: A total of 50 people with CHF (NYHA Class II and III) due to left ventricular systolic dysfunction (33 males; mean age 67 years, 17 females; mean age 68 years) and 30 nominated caregivers recruited from the outpatient departments of two hospitals in the West of Scotland. Sampling was purposive to include patients from a range of CHF severity, ages and sexes. MAIN RESULTS: Managing medications was a responsibility shared by both the patients with CHF and caregivers. Treatment regimens were reported to be difficult to comply with. Health professionals were seen to provide little support for medication management. Pharmacists were viewed as being a good and accessible source of practical assistance who were also knowledgeable about the individual's heart health history. Participants reported valuing advice from pharmacists about the side effects of medications and for their assistance in reducing the complex logistics of medication management and in having medications delivered. CONCLUSIONS: Patients with CHF and caregivers voiced a willingness to try to manage their medication regimen accurately but had a limited capacity to do so. Pharmacists were viewed as providing valuable support to patients with CHF and their caregivers, in terms of medication management. The extended role of pharmacists in medication management of CHF should be encouraged. PMID- 16341954 TI - Pharmacoeconomic evaluation of linezolid versus teicoplanin in bacteremia by Gram positive microorganisms. AB - OBJECTIVE: The objective of this study was to compare the efficiency of linezolid versus teicoplanin in the treatment of bacteremia produced by Gram-positive microorganisms through a pharmacoeconomic analysis based on clinical results obtained from a previous clinical trial. METHODS: We applied an analysis of cost effectiveness elaborated through a pharmacoeconomic model. We defined each unit of effectiveness as 'each successfully cured of infections with bacteremia.' We used the program Pharma-Decision (version Hospital 1.1) that allows to build interactive pharmacoeconomic models. Effectiveness data of both antibiotics were obtained from a published clinical trial, while resources consumed were obtained from the same source and from a consensus provided by a local expert panel. Only direct costs were included in the analysis without taking into consideration indirect costs. The perspective chosen was hospital assistance and the time horizon was set to 28 days. All costs are expressed in Euros. RESULTS: Linezolid demonstrated a better clinical outcome with less associated costs compared to teicoplanin (88.5 versus 56.7% of cured patients and 5,557.04 versus 6,327.43 per treated patient, respectively), thus resulting in a lower cost effectiveness ratio for linezolid versus teicoplanin (6,279.1 versus 11,159.5 per cured patient with a 95% CI of 5,960.2-6,510.4 and 10,865.2-12,647.3, respectively) which results in a the dominant position for linezolid. The sensitivity analysis showed that linezolid was always the most efficient option even when modifying the value of variables with higher uncertainty. CONCLUSIONS: Linezolid is a more efficient option than teicoplanin because it presents higher rate of effectiveness with lower consumption of resources, thus being a dominant alternative in the treatment of Gram-positive infection with bacteremia. PMID- 16341955 TI - Usefulness of the USP Medication Counselling Behavior Guidelines. AB - OBJECTIVE: To assess the perceived usefulness of the United States Pharmacopeia (USP) Medication Counselling Behavior Guidelines among Finnish community pharmacists. The Guidelines were systematically disseminated to pharmacists through a 4-year project (TIPPA 2000-2003). METHOD: A postal questionnaire was sent to a random sample (n = 734) of Finnish community pharmacists (response rate 51%). RESULTS: More than one quarter (27%) of the respondents was aware of the Guidelines. The Guidelines were made known to the respondents mostly via a book on patient counselling skills (41% of those knowing the Guidelines), continuing education (37%), and in-house training (28%). Over 60% of those who knew about the Guidelines regarded them as a good or very good instrument for learning the principles of patient counselling. The Guidelines were considered less useful as an instrument for the self-evaluation of performance in different settings. CONCLUSION: The results of this study indicate that even though the USP Guidelines have been promoted strongly through basic and continuing education during the TIPPA Project, more than half of the respondents were not aware of the Guidelines. The Guidelines were considered more useful in learning the principles of patient counselling than learning self-evaluation of performance. PMID- 16341956 TI - Differences between drug utilisation estimates based on pharmacy sales and on purchases by the resident population. PMID- 16341957 TI - Secondary prevention in coronary heart disease: goal achievement and drug prescribing. AB - OBJECTIVE: To provide an overview of the degree of treatment goal achievement and drug prescribing in patients with coronary heart disease (CHD) in clinical practice. METHOD: Patients undergoing planned or acute percutaneous coronary intervention (PCI) were included. Patients' medical records and the hospital's database on clinical chemistry analyses were studied retrospectively. MAIN OUTCOME MEASURE: Goal achievement and drug prescription relative to published therapeutic guidelines. RESULTS: A total of 200 patients aged 66 +/- 11 years were included. Blood pressure <140/90 mmHg was achieved in 36% of patients 3 months after PCI and total cholesterol <5 mmol/l was achieved in 29% at the time of PCI. Three months after PCI the patients were on 1.4 (range 0-4) drugs with antihypertensive effect (including diuretics). Seventy eight percent were on a statin, 92% were on low-dose aspirin, and 23% had a diuretic in their drug regimen. Five out of 16 patients with diabetes mellitus included in the study were prescribed an angiotensin converting enzyme-inhibitor (ACE-I). CONCLUSION: There is probably a potential for increased degree of goal achievement and for improved drug prescribing in patients with CHD undergoing PCI at our hospital. PMID- 16341958 TI - Fungal chemoprophylaxis with fluconazole in preterm infants. AB - INTRODUCTION: To describe the establishment of a fungal chemoprophylaxis protocol in very low birth weight infants (VLBW). To asses safety, tolerability, and effectiveness of fluconazole administration to these patients. METHOD: According to published scientific evidence (search on MEDLINE 1994-2001) inclusion criteria for the protocol were defined: gestational age <28 weeks and birth weight <1500 g. Fluconazole was given intravenously by infusion. A pharmacotherapeutic follow up of patients included on the protocol was performed for 1 year. RESULTS: Sixteen patients were included on the protocol, two of them died because of causes not related to the drugs given. Significant drug interactions were not observed. Fluconazole side effects were not reported either. None of the patients who finished the chemoprophylaxis showed signs or symptoms of fungal infection. CONCLUSION: Fluconazole chemoprophylaxis in this small number of patients has shown an excellent safety and tolerability profile. The lack of fungal infection points out the possibility of using fluconazole to reduce morbidity and mortality in VLBW. PMID- 16341959 TI - Drug consumption during the 2004 Olympics: the special Olympic Pharmacy. PMID- 16341963 TI - A HIPAA-compliant architecture for securing clinical images. AB - The Health Insurance Portability and Accountability Act (HIPAA, instituted April 2003) Security Standards mandate health institutions to protect health information against unauthorized use or disclosure. One approach to addressing this mandate is by utilizing user access control and generating audit trails of the various authorized as well as unauthorized user access of health data. Although most current clinical image systems [e.g., picture archiving and communication system (PACS)] have components that generate log files for application debugging purposes, there is a lack of methodology to obtain and synthesize the pertinent data from the large volumes of log data generated by these multiple components within a PACS. We have designed a HIPAA-compliant architecture specifically for tracking and auditing the image workflow of clinical imaging systems such as PACS. As an initial first step, we developed HIPAA-compliant auditing system (H-CAS) based on parts of this HIPAA-compliant architecture. H-CAS was implemented within a test-bed PACS simulator located in the Image Processing and Informatics lab at the University of Southern California. Evaluation scenarios were developed where different user types performed legal and illegal access of PACS image data within each of the different components in the PACS simulator. Results were based on whether the scenarios of unauthorized access were correctly identified and documented as well as on normal operational activity. Integration and implementation pitfalls were also noted and included. PMID- 16341964 TI - [Surgical site infections: risk factors and methods of prevention.]. AB - Surgical site infections are the most important problems of surgery. They increase morbidity, mortality, hospital stay and hospital costs. Infection occurring within 30 days after the operation if no implant is left in place or within 1 year if implant is in place is described as surgical site infection. These infections are classified into two categories; incisional or organ/space specific infections. Incisional infections are further divided as superficial or deep incisional infections. Development of surgical site infections is mainly dependent on type of micro-organism and patients' factors related to host resistance. The endogenous flora of the patients' skin, mucous membranes, or hollow viscera is the main source of microorganisms responsible for surgical site infections. Development of such infections can be influenced via a lot of risk factors such as nutritional status, diabetes, smoking, altered immune response, and length of preoperative stay, skin antisepsis, preoperative shaving, duration of operation, antimicrobial prophylaxis, operating room ventilation, inadequate sterilization, the presence of foreign material in the surgical site, and surgical techniques. With the improvement of some of these factors, risk of developing surgical site infections may be decreased. The aim of this study was to discuss the risk factors and prevention measures of surgical site infections in the lightening of current literature. PMID- 16341965 TI - [Does sepsis impair the healing of colonic anastomosis in splenectomized rats?]. AB - BACKGROUND: To investigate the effect of splenectomy on the healing of colonic anastomoses under normal and septic conditions. METHODS: Forty Wistar rats were assigned into six groups: group 1: sham, group 2: colonic anastomose, group 3: splenectomy, group 4: colonic anastomose plus sepsis, group 5: colonic anastomose plus splenectomy, group 6: colonic anastomose plus splenectomy plus sepsis. The rats underwent a standardized left colonic resection and primary anastomosis and/or splenectomy. Sepsis was produced by cecal ligation and puncture. Wound healing was evaluated by bursting pressure and hydroxiproline estimates. RESULTS: Bursting pressures were as follows: group 1: 173 +/- 14 mmHg, group 2: 186 +/- 7 mmHg, group 3: 168 +/- 6 mmHg, group 4: 113 +/- 14 mmHg, group 5: 167 +/- 10 mmHg, and group 6: 183 +/- 3 mmHg. Hidroksiprolin contents were: group 1: 3.5 +/- 0.2 microg/mg, group 2: 3.2 +/- 0.3 microg/mg, group 3: 3.4 +/- 0.2 microg/mg, group 4: 2.3 +/- 0.2 microg/mg, group 5: 3.0 +/- 0.2 microg/mg, grup 6 3.2 +/- 0.1 microg/mg. Statistical significance was found between group 4 and the other groups (p<0.05). CONCLUSION: Sepsis impairs the healing of colonic anastomoses. However, sepsis does not impair the intestinal wound healing in splenectomized rats. PMID- 16341966 TI - The diagnosis of cardiac dysfunction in critically III trauma patients with blunt chest trauma and presumed myocardial contusion: the critical nature of end diastolic volume. AB - BACKGROUND: To evaluate prospectively the effectiveness of monitoring end diastolic volume (EDV) vs pulmonary artery occlusion pressure (PAO) as an estimate of cardiac preload in hemodynamically unstable critically ill 23 myocardial contusion patients with blunt chest trauma admitted to a university trauma center.. METHODS: Patients were resuscitated (ATLS guidelines) with intubation and volume replacement for altered consciousness, hypoxemia or hemodynamic instability. Volume resuscitation and pulmonary artery catheterization were started to measure PAO, EDV, and oxygen utilization parameters. Myocardial performance was determined in both static (0., 24., 48., 72. hours) and dynamic condition after 500cc fluid bolus. RESULTS: A moderate injury severity (GCS 9, TS 11, ISS 34) with MVA mechanism (91%), an average ICU stay of 14 days and a 39% mortality were revealed. Correlation of PAO, EDV with CO, CI demonstrated that EDV is more reliable than PAO However, factoring heart rate into CO determination was more reliable than single preload estimates of EDV PAO-HR, HR-EDV, and HR-PAO. EDV-PAO-HR were correlated more strongly with cardiac output from 24 to 96 hours. A higher AV02 decrease was associated with improved survival, and a nearly significant decrease in EDV. Dynamic comparison found no change in cardiac performance with a small volume challenge. CONCLUSION: Simultaneous consideration of PAO, EDV, HR allowed the most accurate determination of cardiac output. PMID- 16341967 TI - [Approach to the lower gastrointestinal tract bleeding in patients with normal colonoscopic findings.]. AB - BACKGROUND: Lower gastrointestinal bleeding is a commun clinical entity. Although colonoscopic examination is the first choice for diagnosis, it may not be enough to reveal the cause of bleeding in all subjects. METHODS: Eight patients who had massive lower gastrointestinal bleeding having normal colonoscopic findings were retrospectively evaluated at the Department of Trauma and Emergency Surgery, Istanbul University, Faculty of Medicine RESULTS: There were 5 male and 3 female patients with mean age of 51 (28 to 82). Patients received a mean of 13 U (range 2 to 23) with transfused erythrocyte concentrates. Four patients had found to be normal during angiographic, scintigraphy or enterocylytic examinations. Angiography was diagnostic in 4 patients, and identified bleeding from ileocolic pseudoaneurisms (n= 2) branches of jejunal artery (n=2). Embolization procedure were performed in these 4 patients and 3 of them developed intestinal necrosis and underwent surgery. One did not require further treatment. Two of the patients who underwent surgery expired due to sepsis. Patients were hospitalized for a mean of 17 days (range 5 to 37). CONCLUSION: Despite employment of all diagnostic procedures, the cause of bleeding were not detected in half of patients who had normal colonoscopic findings. If angiographic treatment is necessary, superselective arterial embolization should be performed. PMID- 16341968 TI - [The clinical approach to the isolated traumatic hemothorax.]. AB - BACKGROUND: The aim of this study was to evaluate the systematic approach to the isolated traumatic hemothorax. MATERIAL AND METHODS: The treatment modalities of 103 cases with isolated hemothorax was analyzed retrospectively between 1995 and 2003. RESULTS: We observed 103 isolated hemothorax cases. Eighty eight of them were male and 15 were female. The mean age was 39.4 years. The hemothorax was left sided at 41 cases, right sided at 60 cases and bilateral at 2 cases. Fifty three (51,4%) penetrating, forty nine (47,5%) blunt, and one (0,9%) iatrogenic traumas with resultant hemothoraxes were investigated. We performed tube thoracostomy in ninety nine cases. One case required an urgent thoracotomy. We operated twenty one cases and thoracoscopy was performed for six of them. The conservative approach was sufficient for eighty two patients. Mortality was seen in only one case (0,9%), because of a multiorgan failure. The mean rate of the hospital stay was 8,7 days. During operation, we explored for major pulmonary and systemic artery injury. CONCLUSIONS: The decision of exploration or conservative treatment must be made according to the patient stabilization status after the thoracostomy tube insertion. Thoracoscopic evacuation of blood clots from hemothoraxes must be attempted during the early posttraumatic stages, especially in cases refractory to classical drainage methods. PMID- 16341969 TI - [Pediatric head injuries: a retrospective analysis of 280 patients.]. AB - BACKGROUND: To assess etiological factors, clinical features, radiological findings and recovery rates in pediatric head injuries. METHODS: Patients (n =280) with head injuries (age range: 0 - 16 years) hospitalized in Trakya University Department of Neurosurgery between January 1995 and 2004 were analyzed statistically. RESULTS: According to Glasgow Coma Scale (GCS) the patients had minor (GCS: 13- 15 ; 70.1% ), moderate (GCS: 9- 12; 17,1% ), or severe (GCS: 3 to 8; 6,8% ). head injuries The most common etiological factor was fall from a height (34,3%); and the most frequently associated injury was extra-spinal skeletal injury (12,9%). Fifty-one patients (18,2%) underwent neurosurgical operation. 87.5% of them recovered completely, while 12,5% showed partial recovery or died, as graded by Glasgow Outcome Scale (GOS). There was a moderately strong correlation between initial GCS and GOS (r=0,53, p=0,01). CONCLUSIONS: Nearly half of the pediatric head injuries were caused by falls with good prognoses. In the school age, motor vehicle accident (MVA) was the most frequent trauma type. MVA was the most serious type of trauma as demonstrated by its low GCS and GOS scores. Polytraumas, subdural hematomas, cerebral contusions, subarachnoid or intracerebral hemorrhages, cerebral edemas, diffuse axonal injuries, and any cranial lesion which required surgery were found to be related with poor prognosis. PMID- 16341970 TI - An evaluation of civilian aviation accidents in Turkey from 1955 to 2004. AB - BACKGROUND: Documentation and analysis of aviation accidents is very important to improve aviation safety and post-accident survival. In this study we aimed to evaluate the civilian aviation accidents between 1955 and 2003 in Turkey to identify the risk factors for fatal and non-fatal injuries. METHODS: Records of Civilian Aviation Office of The Turkish Ministry of Transportation were retrospectively analyzed. RESULTS: A total of 562 aviation accidents occurred within the boundaries of Turkey in the 48-year study period. There was at least one casualty in 237 of these accidents. There were 27 major accidents with more than 5 casualties. There were more major accidents with survivors than without survivors. Major accidents occurred more frequently within the near vicinity of an airport. There are significantly more accidents with survivors in the accidents occurring near the airports. The mean number of accidents with more than 50 casualties is 0.167 (8/48) per year. CONCLUSION: Accidents occurring near the airports have a more chance of having survivors. Emergency disaster preparedness plans should be on hand and drills with no more than 50 injured should be regularly exercised in all airports. PMID- 16341971 TI - [Crush injuries of the hand.]. AB - BACKGROUND: To carry out a pilot study on the epidemiology of the crush injuries of the hand in our country and to obtain valuable data that will be used for construction of a protective program in the future. METHODS: 124 patients with crush hand injuries were observed retrospectively based on medical records in terms of age, sex, field of work, parts of body affected by the injury, mechanism and underlying causes of the injury RESULTS: Most of the patients were in 0-18 year- age group and inserting the hand into the working machine was the leading cause of injuries. Eighty seven male (70,16% ) and 37 (29,84 %) female patients were included in the study. Most of the patients (44,1%) were working in wood industry at the time of the accident. Interestingly only one hand of all patients were injured. Most commonly injured finger was the middle finger. Most commonly encountered pathology was the tendon laceration in injured fingers. CONCLUSION: Most of the cases were due to lack of attention and education and could be prevented by simple precautions and by wide-spread work education programs leading to a marked decrease in health expenses and loss of man power. PMID- 16341972 TI - [The results of surgical management of ankle fractures.]. AB - BACKGROUND: The results of surgical treatment of ankle fractures and influential factors were evaluated. METHODS: The study ( 1999-2003) included 40 patients (28 men, 12 women) with a mean age of 37.7 years (15-83) who underwent surgical treatment for ankle fractures. The mean follow-up period was 29.6 months (5-66) According to Lauge-Hansen classification, the mechanism of occurence was supination-external rotation in 21 (52.5%), pronation-external rotation in 8 (20%) and supination-adduction in 5 (12.5%) patients, while 6 patients (15 %) were not assessed. Fractures were also classified (Danis-Weber ) as Type A (n= 8; 20%) , B (n = 22; 55%) and C (n =10; 25%) , including 35 (87.5%) bimalleolar fractures. Roberts criteria were used for the final evaluation of the patients. RESULTS: Satisfactory union was obtained in all fractures. According to objective criteria, the results were good in 33 (82.5%) , moderate in 3 (7.5%) and poor in 4 (10%) patients, while corresponding subjective results were obtained in 34 (85%), 3 (7.5%) and 3 (7.5%) patients respectively. CONCLUSIONS: Improved outcomes can be obtained by anatomical reduction and restoration of the ankle joint, and the tilt of talus, fibular shortening and the dysfunction of the syndesmotic joint can be prevented. In our opinion our method of surgical treatment influenced better outcomes obtained. PMID- 16341973 TI - Critical care management of severe head injury in children. AB - BACKGROUND: Our aim was to analyze prognostic factors and their association with outcome among children with severe head injury. METHODS: We conducted a retrospective study among children (n=55) with severe head injury [Glasgow Coma Score (GCS) ?8] who were admitted to our Neurosurgical Intensive Care Unit (ICU) from January 1996 to September 2003. The patients were immediately evaluated with cranial computed tomography (CT) for the severity of head injury as well as for the causes of secondary insults such as hypoxia and hypotension, metabolic and hematological alterations. Outcome analysis was assessed according to Glasgow Outcome Scale Score (GOS) six months after the injury. RESULTS: A poor result occurred in 31 patients (57%) while 24 patients (43%) had favourable results. Multivariate analysis showed significant independent prognostic effect for admission mean systolic blood pressure, presence of hypoxia, multiple trauma, admission GCS score and multiple intracranial lesions (p<0.05). Admission WBC counts and serum glucose levels were not correlated with GOS. CONCLUSION: This study describes clinicoradiologic findings and prognostic factors regarding severe head injury in pediatric patients. The goals of managements of pediatric patients with severe traumatic head injury include normalizing intracranial pressure, optimizing arterial blood gases and systemic blood pressure, and prevention of factors that exacerbate secondary brain injury. PMID- 16341974 TI - Can a pin-tract infection cause an acute generalized soft tissue infection and a compartment syndrome? AB - A patient who developed soft tissue infection and osteomyelitis secondary to pin tract infection after skeletal traction was evaluated. Tibial traction was performed on a patient who had exposed to a femoral pertrochanteric fracture after falling from a tree in a rural public hospital. On the first postoperative day shortly after development of soft tissue swelling, redness, and tenderness in the affected leg, compartment syndrome was noted with subsequent removal of the pin at the same health center. After arrival of the case in our center surgical decompression with an open faciatomy and proper antibiotherapy were instituted. Simultaneously hyperbaric oxygen was administered. After eradication of soft tissue infection we treated the fracture with a Richards compression screw-plate device. The patient was discharged with complete cure. This case presented how seriously a simple pin-tract infection can cause a grave clinical entity resulting in potential loss of an extremity. PMID- 16341975 TI - Intestinal perforation in Wegener's granulomatosis: a case report. AB - Wegener's granulomatosis is a disease characterized by a necrotizing vasculitis and granulomatous inflammation. The localized form involves the upper and/or lower respiratory tracts while in the common generalized form there is a widespread necrotizing vasculitis and renal involvement. Although gastrointestinal involvement which has been detected at necropsy in 24% of the cases is an uncommon finding, it might cause severe complications. We report a patient with clinical Wegener's granulomatosis who subsequently developed gastrointestinal perforation. Gastrointestinal perforation was treated with surgical resection and the patient survived under the treatment of cyclophosphamide and prednisolone with no further gastrointestinal complications. The present case indicates that the gastrointestinal complications might be considered in natural history of Wegener's granulomatosis. PMID- 16341976 TI - Penetrating injury of cranium: a case report. AB - A 38 year-old male patient treated for paranoid schizophrenia for five years was found on a chain saw table at his workplace with a great parasagittal, linear active bleeding wound from left occiput to medial portion of left orbita. He was unconscious.with a Glascow coma score of 5 points as 1-3-1. Cranial radiographies revealed a bone defect from left occipital region to left medial border of orbita. CT scan showed also a great linear tissue damage involving left lateral ventricle, and an intracerebral hematoma located mainly at left frontoparietal region. An emergent left frontoparietal craniotomy was performed. Four centimetres laterally midline, there was a linear, vertical tissue wound. Hemostasis was achieved at first and intracerebral haematoma evacuated. At one week postoperatively, his eyes started to react to verbal commands. At 7 months postoperatively he was attempting to cooperate with eye movements and writing. He was right hemiplegic, aphasic and on the right side deep tendon reflexes were hyperactive. His Karnofsky score was almost 40 points. Cranial injuries due to chain saw accidents are very rare. Early surgical procedures (incl. decompression) combined with aggressive antibiotherapy seem to have a great survival benefits. However best long- term results show that this type of injuries have a great rate of mortality and morbidity despite all surgical and medical treatment procedures. PMID- 16341977 TI - [Gallbladder torsion as a rare cause of acute abdomen; a case report and review of literature.]. AB - Torsion of the gallbladder is a rare cause of acute surgical abdomen. Less than 300 patients have been reported so far. On the other hand incidence seems to be higher than expected especially in elderly patients. This condition must be suspected in an elderly patient who represents signs and symptoms of toxemia and cholecystitis . Diagnosis is usually established during laparotomy. Torsion occurs at the base of the gallbladder around cystic duct and artery. Treatment is emergent cholecystectomy. PMID- 16341978 TI - [Current concepts in neuraxial anaesthesia for labour and delivery]. AB - Epidural analgesia for labour has been associated with an increased rate of motor blockade, and instrumental and Caesarean deliveries. In recent years, these risks were significantly reduced with modern concepts of epidural analgesia, including the use of lower doses of local anaesthetics in combination with opioids. With combinations of 0.0625-0.125 % of bupivacaine plus sufentanil or fentanyl, the incidence of maternal motor blockade approximates 10 % and most parturients are nowadays able to ambulate during labour. Methods of epidural drug administration consist of intermittent boluses, patient-controlled epidural analgesia (PCEA) or continuous infusions. While intermittent top-ups and PCEA do not differ in the amount of local anaesthetics used, continuous infusions have been associated with increases in drug consumption and motor blockade in addition to a higher workload (e. g. frequent adjustments of infusion rates). They therefore do not appear to confer significant benefits during labour analgesia. The most common type of anaesthesia for Caesarean delivery is spinal anaesthesia due to its simplicity, cost-effectiveness and speed of onset. It is suitable for cases of an urgent or emergent Caesarean delivery. General anaesthesia still leads to a higher maternal mortality and should be reserved for absolute emergencies and cases where neuraxial blockade is contraindicated. PMID- 16341979 TI - [The STEMMAT-project as part of health initiative BayernAktiv: adult stem cells from umbilical cord and cord blood as alternative to embryonic stem cell research]. AB - Adult stem cells from umbilical cord and cord blood are an interesting alternative to embryonic stem cells because such research is commonly recognized as ethical undisputed and many aspects are still insufficiently investigated. In the context of the STEMMAT research project (STEM = Stem Cell and MAT = Material) different aspects of stem cells from umbilical cord and cord blood are investigated, to improve basic science understanding and potentially leading someday to a clinical application. PMID- 16341980 TI - [MR imaging of the pelvis in characterization of adnexal masses]. AB - MRI has proved beneficial in characterizing ovarian masses that are indeterminate by sonographic criteria. MRI can distinguish between benign and malignant ovarian tumors with high sensitivity and specificity and is capable of characterizing many adnexal masses. MRI should be therefore included in the preoperative diagnostic of adnexal masses. PMID- 16341981 TI - [Suspicion for intraabdominal adhesions -- is open laparoscopy the gold standard?]. AB - Laparoscopy is used for most surgical procedures in gynaecology. In general complications are rare. However, one of the most critical steps is the initial laparoscopic entry into the peritoneal cavity. According to the literature serious complications occur in approximately 1-2/1 000 cases. Whereas major vascular injuries are mainly recognised immediately, delayed recognition of bowel injuries is frequent. Complication rates of different entry procedures used in gynaecological laparoscopy are similar even in high risk patients (intraperitoneal adhesions, obesity). Utilising an open - instead a closed - entry (either by Veress needle or first trocar) technique or alternativ entry positions are suggested by some authors. This review presents data available in the literature and highlights that open laparoscopy is no gold standard. PMID- 16341982 TI - Developing an oncology portfolio of anticancer drugs: the experience of one pharmaceutical company. AB - For a company or institute that wishes to develop new anti-cancer agents, it is necessary to establish a portfolio of agents to reduce the risk of failure. Success rates for developing new agents are low and therefore different biological effect areas should be explored to ensure that at least one agent targeting cancer or a specific histological sub-type of cancer is effectively developed. This paper describes how one pharmaceutical company has developed a range of different agents with the ultimate aim of developing as many of these as is technically feasible into useful new medicines for the treatment of cancer. PMID- 16341983 TI - [Diagnosis and therapy of breast cancer--meeting highlights ASCO 2005]. PMID- 16341984 TI - The congruity between urine and cervical swab cultures in pregnant women with versus without amniotic fluid infection. AB - INTRODUCTION: We observed the congruity of bacteria found in urine and cervix of pregnant women with amniotic fluid infection (AFI) compared to healthy controls. PATIENTS AND METHODS: Over three years, we prospectively analysed urine and cervical swabs cultures in 120 pregnant women in gestational week 16-20. The patient population was divided in two groups: group I patients had clinical symptoms of AFI. The rest of the patients were designated as healthy controls (group II). Congruity between findings in both groups was observed. In patients with bacterial growth, antibiotic treatment was initiated as recommended and once ended - culture probes were repeated. RESULTS: The rates of nonsignificant and significant bacteriuria were doubled in group I. Fifty-five percent (n = 33) of patients in group I had identical bacteria cultured both from the urine sample and cervical swab, in contrast to only 13.3 % in group II. Congruity was most pronounced for Klebsiella species and E. coli, the later being single most dominant isolate in regards to both cultures. After antimicrobial treatment, microbial eradication occurred in 15 patients (45.5 %). The observed incidences of abortions and preterm deliveries were significantly lower in patients with microbial eradication versus patients with microbial persistence. CONCLUSION: Patients with clinical symptoms of AFI have high risk for bacteriuria identical to bacterial culture from cervical swab. Antimicrobial treatment was effective only partially where indicated. Screening for eradication is recommended and consensus on the most appropriate therapy is needed. PMID- 16341985 TI - [MR-guided vacuum assisted breast biopsy]. AB - OBJECTIVE: This prospective study was undertaken to determine the feasibility of MR-guided vacuum assisted breast biopsy with the Vacora-vacuum-biopsy system for histological evaluation of suspicious lesions in MR-mammography. MATERIAL AND METHODS: During 3 months MR-guided vacuum assisted breast biopsy was indicated in 12 patients with suspicious lesions in MR-mammography. RESULTS: MR-guided vacuum assisted breast biopsy with the Vacora-vacuum-biopsy system could be performed in 9 of 12 patients. In 2 patients the lesions could not be identified at the time of the intervention. In one patient the intervention could not be performed due to obesity. Histopathology revealed benign lesions in 8 patients and malignancy in one patient. In one of the cases with benign histology, the biopsy specimen was not representative for the lesion. CONCLUSIONS: MR-guided vacuum assisted breast biopsy with Vacora-vacuum-biopsy is technical feasible can be performed with a low complication rate. PMID- 16341986 TI - [Breast cancer -- plastic surgical strategies for the treatment of tumour infiltration of the thoracic wall and brachial plexus]. AB - Extensive locoregional recurrency or metastatic involvement of brachial plexopathy is a condition that is often associated with advanced systemic breast cancer. In the past the role of surgeon was restricted due to a scepticism as to whether any benefit will be provided for the patient. In the recent 25 years however safe and refined plastic surgical approaches have been developed that provide more options to treat even complex recurrent disease. The strategy of plastic surgery in an interdisciplinary approach of gynecology, oncology and radiotherapy as well as thoracic surgery is outlined. PMID- 16341988 TI - Stroke acute management and recovery. PMID- 16341990 TI - Acute stroke treatment in the community: improving our performance and expanding our options. AB - Three-quarters of all strokes are managed outside of academic medical centers, making optimization of stroke care in the community setting a central part of minimizing the disability from stroke. In addition, the care of stroke patients crosses multiple different components of the health care system and requires a multifaceted and coordinated approach. This article reviews the current status of stroke care across the care continuum and reviews methods found to be effective in educating the public and organizing regional delivery of hyperacute stroke care. Successful community-based stroke care models and current national initiatives in improving hospital management are discussed. Local implementation of these strategies provides an exciting opportunity to improve outcomes after stroke. PMID- 16341991 TI - Neuroimaging advances and the transformation of acute stroke care. AB - Neuroimaging techniques have transformed the delivery of stroke care. Multimodal computed tomography and magnetic resonance imaging rapidly illustrate the vascular and parenchymal correlates in acute ischemic and hemorrhagic stroke. Optimal selection of thrombolytic candidates and the characterization of ischemic sequelae may be achieved with imaging. The nature and topography of intracerebral hemorrhage may also be defined. Increasing use of multimodal imaging in acute stroke has expanded our current understanding of stroke pathophysiology and streamlined the care of stroke patients from the hyperacute to chronic phases. The integration of neuroimaging techniques in research studies has elucidated pitfalls in the translation of novel therapy into clinical practice. This review explores the impact of neuroimaging advances in stroke and emphasizes the critical role of these modalities in the care of patients with ischemic and hemorrhagic events. PMID- 16341992 TI - Transient ischemic attack: a dangerous Harbinger and an opportunity to intervene. AB - Transient ischemic attack (TIA) and ischemic stroke are both characterized by sudden onset of neurological symptoms due to focal cerebral ischemia, but they are distinguished by the duration of neurological symptoms, with TIA traditionally defined by resolution of symptoms within 24 hours and stroke reserved for symptoms of longer duration. Because TIA and ischemic stroke share etiologies, it is not surprising that the recommended evaluations and secondary prophylaxis are identical. However, recent studies suggest that optimal management of TIA and stroke may differ more than previously recognized. The short-term risk of ischemic stroke after TIA is very high, which may be because rapid recovery from neurological symptoms is indicative of reversal of ischemia and tissue still at risk. Regardless of whether residual symptoms or infarction are present, rapid recovery appears to predict a greater short-term risk of subsequent ischemic stroke and is more likely to reflect a distinct, unstable pathophysiology. Therefore, it may be more useful to characterize acute ischemic cerebrovascular syndromes on the basis of the extent of rapid recovery (i.e., on the inferred reversal of ischemia) than on the completeness of recovery at 24 hours. Patients with substantial rapid recovery may be those for whom acute intervention is most warranted. PMID- 16341993 TI - Patent foramen ovale and stroke risk: incidental finding or causal culprit? AB - Between 17 and 27% of the population has a patent foramen ovale (PFO) based on autopsy studies. Given these percentages, a PFO could be considered a variant of normal cardiac anatomy and not an anomaly. As this anatomy is so prevalent in the normal population, it is likely that there are other factors involved in patients who have a PFO and a stroke. This review will evaluate the existing literature on PFO and stroke to determine what constitutes an increased risk of embolic events in patients with PFO. Given the paucity of existing literature, it is clear from this review that there is a vital need for randomized clinical trials to answer important questions, including what constitutes high-risk anatomy and optimal treatment for patients for the prevention of recurrent stroke. PMID- 16341994 TI - Medical management of carotid artery disease. AB - It is estimated that more than 700,000 strokes occur in the United States annually, and stroke remains the third leading cause of death. As many as 20% of strokes are due to carotid artery disease. Approaches to therapy include surgery and stenting in appropriate settings, but medical therapy is a critical aspect of management of carotid artery disease. Best medical therapy for carotid stenosis has evolved in recent years and clinicians can now employ newer antiplatelet agents, aggressive use of statins, and targeted blood pressure reduction. Application of this multimodal cocktail holds promise for reducing vascular events in patients with carotid artery disease and may reduce the need for revascularization procedures. PMID- 16341995 TI - Arm function after stroke: from physiology to recovery. AB - There are varying degrees of spontaneous improvement in arm paresis over the first 6 months after stroke. The degree of improvement at 6 months is best predicted by the motor deficit at 1 month despite standard rehabilitative interventions in the ensuing 5 months. Animal studies indicate that the loss of fine motor control, especially individuation of the digits, is due to interruption of monosynaptic corticomotoneuronal connections. Spasticity occurs because of loss of cortical modulatory control on descending brain stem pathways and spinal segmental circuits but is not a major cause of motor dysfunction. Quantitative studies of reaching movements in patients suggest that arm paresis consists of higher-order motor planning and sensorimotor integration deficits that cannot be attributed to weakness or presence of synergies. Cortical stimulation experiments in animals and functional imaging studies in humans indicate that motor learning and recovery after stroke share common brain reorganization mechanisms. Rehabilitation techniques enhance learning-related changes after stroke and contribute to recovery. Future research will benefit from using quantitative methods to characterize the motor impairment after stroke and by applying concepts in motor learning to devise more physiologically based rehabilitation techniques. PMID- 16341996 TI - Depression and stroke: cause or consequence? AB - Depression after stroke is common. Although different opinions exist about the definition, diagnosis, and measurement of outcomes related to depression after stroke, there is little debate about the prevalence of depression symptoms and their impact on stroke survivors and their families. Depression after stroke has long been recognized as a common condition with many negative effects in the poststroke period, but more recently depression has also been identified as an independent stroke risk factor. Given that there are at least 500,000 new ischemic strokes yearly in the United States, a conservative estimate is that 150,000 U.S. stroke survivors develop poststroke depression each year. Because effective treatments exist but are likely underutilized for depression, this is an important example of an evidence-practice gap to which increased efforts to improve care should be made. Such efforts would likely improve not only patient symptoms but may also decrease stroke risk, influence stroke functional recovery, decrease mortality, and reduce poststroke health care utilization. This article provides an overview of depression diagnosis in stroke, reviews the epidemiology of poststroke depression and its associated morbidity and mortality, and reviews existing evidence on the treatment and prevention of poststroke depression. PMID- 16341997 TI - Pediatric stroke: what do we know and what do we need to know? AB - Stroke is a heterogeneous disorder and an important cause of mortality and chronic morbidity in children. Estimates of international incidence rates for childhood stroke have varied widely. Arterial ischemic stroke is reported to be more common than hemorrhagic stroke in children. The clinical presentation of stroke in children differs according to the child's age and stroke type and location. Several risk factors for ischemic and hemorrhagic stroke in children have been reported and include cardiac disorders, blood disorders, vasculopathies, viral infections, and arteriovenous malformations. Current treatment recommendations for stroke in children are based on small nonrandomized trials, adult stroke studies, case series, or consensus or individual expert opinion. Over half of children with stroke will develop lifelong cognitive or motor disability, and up to a third will have a recurrent stroke. International studies have provided important information on stroke in children, but major gaps in our knowledge of the disorder still exist. Currently, there is a need for prospective cohort studies in diverse populations, which utilize a consensus pediatric stroke classification system and a standard evaluation of risk factors and outcome, so that treatment and prevention strategies can be developed. PMID- 16341998 TI - Perinatal arterial stroke: understanding mechanisms and outcomes. AB - Arterial ischemic infarction occurring around the time of birth is an increasingly recognized cause of neurological disability in children. The rate of arterial infarction in neonates is as high as the annual incidence of large vessel ischemic stroke in adults. Factors contributing to this increased risk of stroke among neonates include complications that occur before, during, and after delivery. Maternal conditions that have been associated with perinatal stroke in the fetus include prothrombotic disorders, cocaine abuse, and placental complications such as chorioamnionitis and placental vasculopathy. In many cases, the placenta is suspected to be the underlying embolic source for perinatal stroke, although data on placental pathology is often lacking. During the delivery process, an infant may develop a cervical arterial dissection that leads to stroke. Several conditions in the neonatal period predispose to perinatal stroke including prothrombotic disorders, congenital heart disease, meningitis, and systemic infection. Perinatal stroke may present with neonatal seizures during the first weeks of life or may be asymptomatic until months later when the infant is first noted to have pathological handedness. The outcome of perinatal stroke is variable and depends on severity, anatomic localization, and other factors not yet well characterized. As many as 50% of infants with documented stroke recognized in the newborn period do not develop a hemiparesis. The incidence, clinical presentation, pathogenesis, risk factors, and outcome of this increasingly recognized disorder are reviewed. PMID- 16341999 TI - Subarachnoid hemorrhage and inflammation: bench to bedside and back. AB - Subarachnoid hemorrhage (SAH) is a devastating and complicated disease. The development of therapeutic interventions has been hampered by a poor understanding of the three components of the disease pathology in SAH: aneurysm rupture, cerebral edema, and vasospasm. The role of inflammation in the pathology of subarachnoid hemorrhage will be reviewed. The events leading up to aneurysm rupture are heralded by degradation of the endothelial cell layer integrity and inflammatory cell infiltration into the wall of the aneurysm. This is associated with release of active agents that can digest the basement membrane and may cause rupture. After rupture, cytokine release by mononuclear leukocytes is associated with early edema. Vasospasm is a complicated process that includes arterial wall thickening and vasoconstriction. Evidence supports the role of inflammation in free radical formation and in perturbations in nitric oxide and endothelin-1 levels that are important mediators of the vasoconstriction in vasospasm. Targeting the inflammatory mediators associated with the three prominent events in SAH is a promising strategy for reducing the mortality and morbidity in these patients. More study is needed to determine which specific effectors in the inflammatory cascade may serve as targets for intervention. PMID- 16342000 TI - Treatment and prevention of primary intracerebral hemorrhage. AB - Intracerebral hemorrhage (ICH), which constitutes 10 to 15% of all strokes and affects approximately 65,000 people each year in the United States, has the highest mortality rate of all stroke subtypes. Hypertension, cerebral amyloid angiopathy, and anticoagulation underlie the majority of cases of ICH. Warfarin not only increases the risk but also increases the severity of ICH by causing hematoma expansion. With the advent of gradient-echo magnetic resonance imaging, patients with underlying cerebral amyloid angiopathy or hypertensive vasculopathy can be identified, and measures can be taken to prevent ICH. Initiating an antihypertensive regimen in a patient with nonlobar microbleeds suggestive of hypertensive vasculopathy, and withholding warfarin in patients with lobar microbleeds suggestive of cerebral amyloid angiopathy, are emerging prevention strategies. Although a treatment for cerebral amyloid angiopathy does not exist, agents targeting beta-amyloid metabolism and bioactivity are promising candidates. Strategies for preventing warfarin-associated hemorrhage include strict monitoring of anticoagulation levels and using agents such as direct thrombin inhibitors. The future of ICH management lies in therapies targeted at the pathophysiological steps in ICH. Potential treatments include glutamate receptor antagonists for preventing glutamate excitotoxicity, matrix metalloproteinase and thrombin inhibitors for preventing perihematomal edema, and recombinant activated factor VII for preventing hematomal expansion. PMID- 16342001 TI - Inhibition of P-glycoprotein function by several antidepressants may not contribute to clinical efficacy. AB - INTRODUCTION: In many depressive patients the negative feedback mechanism of the HPA (hypothalamic-pituitary-adrenocortical) axis is impaired. It has been suggested that antidepressants inhibit membrane glucocorticoid transporters like P-Glycoprotein (Pgp) and hence enhance the intracellular glucocorticoid concentration, leading to an increased glucocorticoid-receptor mediated gene transcription and therefore to normalization of the function of the HPA axis. The aim of this study is to investigate inhibition of Pgp by several different antidepressants. METHODS: We characterized the inhibitory potencies of the antidepressants in two in vitro assays by using calcein-AM as Pgp substrate. The two different cell-systems expressing Pgp were: 1. PBCEC (porcine brain capillary endothelial cells) as model for the blood-brain-barrier, and 2. A human lymphocytic leukaemia cell line CEM and the multi-drug-resistant (MDR) cell line VLB-100, expressing Pgp as model for the human protein. RESULTS: All of the antidepressants tested inhibit the transport of calcein-AM by Pgp in the micromolecular range. DISCUSSION: Because this inhibition is only seen at concentrations above therapeutically relevant plasma levels, their effect my not play a role for the mechanism of action of the antidepressants tested. PMID- 16342002 TI - Psychological effects of (S)-ketamine and N,N-dimethyltryptamine (DMT): a double blind, cross-over study in healthy volunteers. AB - INTRODUCTION: Pharmacological challenges with hallucinogens are used as models for psychosis in experimental research. The state induced by glutamate antagonists such as phencyclidine (PCP) is often considered as a more appropriate model of psychosis than the state induced by serotonergic hallucinogens such as lysergic acid diethylamide (LSD), psilocybin and N,N-dimethyltryptamine (DMT). However, so far, the psychological profiles of the two types of hallucinogenic drugs have never been studied directly in an experimental within-subject design. METHODS: Fifteen healthy volunteers were included in a double-blind, cross-over study with two doses of the serotonin 5-HT2A agonist DMT and the glutamate N methyl-D-aspartate (NMDA) antagonist (S)-ketamine. RESULTS: Data are reported for nine subjects who completed both experimental days with both doses of the two drugs. The intensity of global psychological effects was similar for DMT and (S) ketamine. However, phenomena resembling positive symptoms of schizophrenia, particularly positive formal thought disorder and inappropriate affect, were stronger after DMT. Phenomena resembling negative symptoms of schizophrenia, attention deficits, body perception disturbances and catatonia-like motor phenomena were stronger after (S)-ketamine. DISCUSSION: The present study suggests that the NMDA antagonist model of psychosis is not overall superior to the serotonin 5-HT2A agonist model. Rather, the two classes of drugs tend to model different aspects or types of schizophrenia. The NMDA antagonist state may be an appropriate model for psychoses with prominent negative and possibly also catatonic features, while the 5-HT2A agonist state may be a better model for psychoses of the paranoid type. PMID- 16342003 TI - Enhanced ROS-generation in lymphocytes from Alzheimer's patients. AB - INTRODUCTION: Reactive oxygen species (ROS) have been implicated in neurodegeneration and seem to be involved in the physiology and pathophysiology of several diseases, including normal aging and Alzheimer's disease (AD). Enhanced ROS production in aging or AD is not restricted to the brain, but can also been seen in several peripheral tissues. The objective of the present study was to evaluate whether the mechanisms involved in the generation of oxidative stress in normal senescence and Alzheimer's disease are identical or not. METHODS: We analysed intracellular basal levels of ROS in lymphocytes from AD patients and healthy young and aged not-demented subjects as well as ROS levels following stimulation with d-ribose and staurosporine in all three groups. ROS levels were measured by flow cytometry using the intracellular fluorescence dye dihydrorhodamine123 (DHR123). RESULTS: Our study shows that AD lymphocytes have increased basal levels of ROS, low susceptibility to ROS stimulation by 2-deoxy- D-ribose (dRib) and an increased response to staurosporine when compared with age matched controls. DISCUSSION: The data suggest that the defect(s) responsible for enhanced ROS production in AD may involve different or additional biological pathways than those involved in enhanced ROS generation during aging. PMID- 16342004 TI - Prolonged confusional state following electroconvulsive therapy--diagnostic clues from serial electroencephalography. AB - INTRODUCTION: Cognitive impairment occasionally occurs after electroconvulsive therapy (ECT) and usually resolves within a few days. Any prolonged cognitive alterations or confusional states may have additional causes and require extensive diagnostic effort. Since cognitive dysfunction can also be caused by ictal states, electroencephalography (EEG) is an essential tool for these conditions. METHODS: We report on a female patient with pharmacotherapy resistant major depression who had been treated by a series of ECT and subsequently developed severe confusion and fluctuating amnesia. RESULTS: Laboratory and neuroimaging examinations were normal, however, EEG revealed a severe intermittent slowing with rhythmic high amplitude delta-/theta-activity and sporadic bitemporal sharp waves. Oral application of 1 mg lorazepam led to a sudden improvement of EEG abnormalities. Consequently, non-convulsive status epilepticus (NCSE) was suspected and the patient was regularly treated with lorazepam, accordingly. Clinically the confusional and amnesic symptoms declined, whereas serial EEG recordings showed a further improvement and normalization of brain electric activity. CONCLUSION: Routine EEG is an indispensable tool in patients with sudden deterioration of cognitive functions and unclear neuropsychiatric symptomatology. A testing dose of lorazepam can help to classify EEG abnormalities in terms of ictal EEG patterns. PMID- 16342005 TI - Effects of lithium on saccadic eye movements in healthy subjects in a ten-day double-blind placebo-controlled cross-over pilot study. AB - INTRODUCTION: Although previous studies have shown that lithium modifies eye movements or psychomotor speed, no studies have ever explored the predictive saccades or memory guided saccades during lithium administration. We took the objective to determine the influence of lithium in pseudo-random, predictive or memory-guided saccades in healthy subjects with a view to detect reduced psychomotor speed, inability to anticipate incoming events, or working memory deficits. METHODS: A ten day lithium-placebo randomized double-blind cross-over pilot study was carried out with 12 healthy male volunteers. The cognitive assessment included pseudo-random, predictive and memory guided saccades before and after lithium and placebo periods. A biological assay substantiated the lithium effect on TSH and thyroid hormones. RESULTS: There was no change in pseudo-random or memory guided saccades when comparing lithium or placebo administration. However the ratio of anticipated saccades decreased under the lithium sequence while it remained stable under placebo. Also, subjects having lithium serum levels of > 0.5 meq/l had longer latencies in anticipated saccades. CONCLUSION: The findings do not support a major effect of lithium on alertness or on working memory, although the dosage and duration of lithium was sufficient to modify TSH blood level. Nevertheless, lithium treatment was associated with decreased anticipation in predictive saccades, suggesting this could reflect a reduced ability to anticipate quick motor movements and could be related to the well-known effect of lithium as an anti-impulsive medication. PMID- 16342006 TI - Galactorrhea during treatment with trimipramine. A case report. AB - Trimipramine, a tricyclic antidepressant, faced renewed attention in the last decade for its well-established antidepressant as well as its sleep-promoting properties. While the antidepressant mode of action of trimipramine is still unclear, its antihistaminic and anticholinergic potency is far better known, leading to side effects such as somnolence or dry mouth. Elevated prolactin serum levels, caused by antidopaminergic action, have been repeatedly observed. However, to our knowledge no case of galactorrhea while under treatment with trimipramine had been previously reported. A 31-year-old female inpatient, admitted to our hospital after an attempted suicide, was treated initially with paroxetine 20 mg/d. Because of persisting insomnia, after 5 weeks we gave a supplementary medication, trimipramine, with a successive increase in dose up to 150 mg/d. After another 9 weeks the patient reported a strong tension in both breasts together with galactorrhea. The prolactin serum level was 21.8 ng/ml. After reduction of the trimipramine dose to 50 mg/d, the symptoms disappeared within a few days. We discuss our case with special regard to the combination of trimipramine with a selective serotonin reuptake inhibitor (SSRI). This commonly used comedication could have led to an aggravation of the problem because (1) most SSRIs stimulate prolactin secretion and (2) trimipramine is metabolized by the cytochrome P450 2D6, which in turn is inhibited by all SSRIs, thus potentially leading to elevated trimipramine plasma levels. The problem can probably be kept to a minimum by giving lower doses of trimipramine (up to 50 mg/d) if co-administered with an SSRI. PMID- 16342007 TI - Exacerbation of migraine attacks during treatment with the selective serotonin reuptake inhibitor sertraline. A case report. AB - Abnormal signal transmission in central serotonergic pathways is supposed to play an important role in the pathogenesis of migraine and major depression. We report on a patient, who was treated during an episode of depression with the selective serotonin reuptake inhibitor (SSRI) sertraline and developed frequent migraine attacks under this therapeutical regime. Single migraine attacks were treated successfully with triptanes. Although SSRIs may be beneficial for migraine prophylaxis at long term administration, this case suggests that acute administration of SSRIs in migraineurs may include the risk of worsening migraine. PMID- 16342008 TI - Guillain-Barre syndrome after septicemia following clozapine-induced agranulocytosis. A case report. AB - We report the case of a patient with schizophrenia, who experienced agranulocytosis during clozapine treatment, followed by bronchopulmonal infection and Guillain-Barre syndrome. The case was recorded within the German surveillance project "drug safety in psychiatry" (AMSP). PMID- 16342009 TI - Electroconvulsive monotherapy in the treatment of acute confusion psychosis. A case report. AB - This report focuses on the successful treatment of a most acute case of confusion psychosis according to the concept of Karl Leonhard. The 18-year-old patient was hospitalized three times before the current episode and his case has been characterized as pharmacologically treatment-resistant psychosis since he failed to respond to any psychopharmacological therapy including sufficient clozapine medication. In the patient's history, typical and atypical antipsychotic as well as mood-stabilizing and sedating pharmacological treatments have been conducted. However, only adverse effects could be observed. When receiving electroconvulsive monotherapy (ECT), the patient showed a marked reduction of symptoms while experiencing no adverse effects. The implications of this finding are discussed with regard to Leonhard's diagnostic system. PMID- 16342010 TI - [Physical (in)activity in childhood and adolescence]. AB - In the last two decades in Germany and other countries physical activity of children and adolescents declined while sedentary lifestyle increased. Physical inactivity in childhood and youth is related with coronary heart disease risk factors and higher prevalence of obesity. International recommendations suggest daily 60 minutes or more moderate to vigorous physical activity and less than 2 hours sedentary activity in leisure. PMID- 16342011 TI - [A common and insidious side-effect: allergic contact dermatitis caused by bufexamac used in the treatment of dermatitis. Results from the Information Network of Departments of Dermatology (IDVK)]. AB - BACKGROUND AND OBJECTIVE: Bufexamac is a non-steroidal, anti-inflammatory drug used in the topical treatment of atopic dermatitis, stasis dermatitis and perianal eczema. The substance is known to cause severe allergic contact dermatitis (ACD) as an adverse effect (AE), which may be indistinguishable from the eczema which is to be treated. Hence the diagnosis of this AE is often considerably delayed. In order to estimate the quantitative importance of ACD to bufexamac, data of the Information Network of (German) Departments of Dermatology (IVDK) from July 1999 to December 2004 were analysed. PATIENTS AND METHODS: During the study period, 39,392 unselected patients from 40 German departments of the IVDK were patch tested with bufexamac (5 % pet). The results of the reading after 72 hours were analysed. The dichotomized patch test result was further assessed for possible risk factors from the patients' history and clinical diagnosis by Poisson regression analysis. RESULTS: In 560 of 39,392 patients contact allergy to bufexamac was diagnosed, i. e. 1.4 % (95 % confidence interval: 1.3 - 1.5), standardized for sex and age. The Poisson regression analysis revealed a significantly increased risk associated with the following factors: multiple sensitization, perianal eczema, underlying atopic dermatitis, leg dermatitis, female gender and residence in areas of Germany other than Eastern Germany. The latter observation can be explained by low prescription rates in Eastern Germany. CONCLUSION: Bufexamac is an important allergen. Extrapolating the frequency of 1.4 % in our data to the whole German population by the CE-DUR approach yields an estimate of about 6000 cases per year. In view of the high frequency of sensitization, the pitfalls in diagnosis, the severity of the course of disease and the lack of efficacy of this drug, the risk to benefit ratio is obviously critical. PMID- 16342012 TI - [Forgoing life-sustaining measures in patients at the end of life in the German speaking part of Switzerland: results of a death certificate study]. AB - BACKGROUND AND OBJECTIVE: There have not been any comprehensive data from German speaking countries on the medical practice of withholding or withdrawing treatment at the end of life. This study from the German-speaking part of Switzerland provides the first in-depth analysis in this field. This study is based on data from this region and is a contribution to a large empirical research project on medical end-of-life decisions in 6 European countries (EURELD). METHODS: Continuous random samples (n = 4991) were taken from all deaths in the German-speaking part of Switzerland that had been reported to the Swiss Federal Office of Statistics between June and October 2001. Doctors who had been attending the deceased person were asked to complete mailed questionnaires, their replies being kept strictly anonymous. RESULTS: The response rate was 67 %. Medications were withheld or withdrawn in 48 % of all treatments forgone: among these, antibiotics accounted for 17 %. Other potentially life-sustaining medical measures forgone included artificial hydration (12 %), surgery (7 %), artificial feeding (6 %), chemotherapy (6 %), diagnostic tests (4 %), hospital admissions (3 %), renal dialysis (2 %), blood product infusions (2 %), intubation (2 %), ventilation (2 %), resuscitation (2 %), and radiotherapy (1 %). 43 % of all treatments were forgone in patients who died in hospital, 42 % in nursing homes, and 15 % at home. In almost three-quarters (73 %) of the treatments forgone, a primary-care doctor had ordered the treatment to be withheld or withdrawn. On average, forgoing treatment led to a life-shortening effect of more than one month in 8 % of all cases. The proportion was higher for renal dialysis (25 %), blood products infusion (18 %), and diagnostic tests (16 %). CONCLUSIONS: Forgoing life-sustaining medical treatment comprises a wide range of decisions taken in many different clinical settings. In most cases the likely lifespan is only slightly shortened. PMID- 16342013 TI - [Primary diagnosis of an interrupted aortic arch in a 65-year old woman with hypertension]. AB - HISTORY AND PHYSICAL EXAMINATION: A 65-year-old woman presented in a hypertensive crisis and with angina pectoris. She had a history of hypertension for several years and medication included five different antihypertensive drugs. On physical examination a faint systolic murmur was heard. Weak femoral pulses were felt, but not the arterial pulses distal to the groin. INVESTIGATIONS: An attempt to perform coronary arteriography failed because it was impossible to pass the catheter across the aortic arch. MR-angiography and cardiac catheterization via the brachial artery confirmed the diagnosis of a complete interruption of the aortic arch distal to the left subclavian artery and showed distinct collateral circulation predominantly via the internal mammary arteries. DIAGNOSIS AND THERAPY: Because of the age of the patient we assumed that the pathogenesis of this interruption of the aortic arch probably was progression and finally occlusion of an aortic coarctation. It was not possible to distinguish the findings from a true congenital atresia, because there was no histological examination. The patient rejected surgery and conservative therapy with frequent monitoring seemed justified, considering the good blood pressure adjustment and the extensive collateral vascularization. CONCLUSION: Patients with complete interruption of the aortic arch very rarely do reach late adult age without previous surgical intervention. Indeed, such a situation is only conceivable when there is a good collateral blood supply and no concomitant shunt defects. There is an indication for operation, but as there are insufficient follow-up data this should be weighed up carefully in elderly patients whose blood pressure is well controlled. PMID- 16342015 TI - [Management of erectile dysfunction in the cardiac patient]. PMID- 16342016 TI - [Oxidative stress and cardiovascular diseases]. AB - A number of diseases like hypercholesterolemia and atherosclerosis, hypertension, congestive heart failure, diabetes, ischemia-reperfusion, neurodegenerative diseases as well as acute and chronic inflammatory diseases are characterized by an increased steady-state concentration of reactive oxygen species (ROS). On a biomolecular level an enhanced oxidative stress causes damage of proteins, lipids and nucleic acids. Both the experimental and therapeutic efficiency of different antioxidative compounds (like various antioxidative enzymes) , drugs, metabolites and vitamins for the maintenance of an appropriate intracellular redox potential underline the importance of an excessive ROS-formation for these diseases. Control of excessive ROS-formation can be obtained by angiotensin converting enzyme (ACE-) inhibitors, by AT (1)-receptor blockers, by statins and other lipid lowering compounds, by improved expression of antioxidative enzymes (superoxide dismutase, catalase etc.), by compounds such as probucol, certain vitamins, pyruvate, by lipid apheresis and by physical exercise training, which displays surprising efficacy. PMID- 16342017 TI - [Caution in foregoing patient education]. AB - A key part of the individual patient rights is the renunciation of explicit information on a planned surgical procedure. However, jurisdiction has assigned strict requirements for the validity of such a denial. To ensure legal validity of the denial, the surgeon has to obtain informed consent signed by the patient stating that the patient rejected detailed medical information, but did understand the nature of the planned procedure. The initiative to receive just limited medical information must be taken by the patient himself, not the surgeon. Thus, the patient may choose to receive either detailed or limited information on the surgical procedure. Important to know: A limited time schedule of the surgeon is not a valid justification for a limited informed consent. PMID- 16342018 TI - [What therapy options are reliable in smoke inhalation?]. PMID- 16342019 TI - [Migraine: effective therapy with acupuncture? Comment: needles in migraine: not where, but why! On the contribution in DMW 24/2005]. PMID- 16342021 TI - [Scientific internet research: advanced possibilities provided by new search engines]. PMID- 16342022 TI - [Using content analysis in cross-cultural psychosomatic research]. AB - The Gottschalk-Gleser-analysis, a content-analytical technique, measures anxious and aggressive affects. For decades this technique was primarily and widely implemented in psychosomatic research. Today, this technique is offered in numerous languages. Since this technique is time consuming in application and analysis, computer software has been developed for English (Psychiatric Content Analysis and Diagnosis, PCAD 2000) as well as for German contents (Dresdner Angstworterbuch, DAW). This study compares the anxiety scale of both versions in order to evaluate their comparability in cross-cultural psychosomatic studies. The English and German version of 96 different abstracts of the German Journal for Psychotherapy, Psychosomatic Medicine and Medicine Psychology from 2003 and 2004 with a total of 31 000 words were compared. The scores of the Gottschalk Gleser anxiety scales of both computer programs correlates between 0.21 and 0.59 (p < 0.05, only exception: shame anxiety). The means of the scores calculated by the PCAD and DAW significantly differ for all scales. Therefore, these two content analytical methods DAW and PCAD provide only limited use for cross cultural comparisons in psychosomatic research. PMID- 16342023 TI - [The Relationship Patterns Questionnaire (RPQ): a validation using a clinical sample]. AB - The Relationship Patterns Questionnaire RPQ was designed to assess the relationship patterns concerning the individual's own behavior as well as the anticipated behavior of an important reference person. In this study, the questionnaire is examined psychometrically by using a sample of psychosomatic inpatients (N = 160). The underlying circumplex structure could be confirmed by means of factorial analyses as well as by the correlations between IIP and RPQ. There were no significant correlations between physical complaints (GBB) and relationship patterns; however, a less affectionate relation was associated with greater psychosocial strains (SCL-90-R) both on the self and on the object level. The pre-post analysis revealed a tendency to a greater ability to assert oneself as well as to be open opposite the reference person, but the effect size was small. Possibilities and limits of the present RPQ version are discussed and conclusions are drawn concerning the further development. PMID- 16342024 TI - [The validity of the depersonalisation-derealisation scale of the narcissism- inventory]. AB - The German Narcissism-Inventory contains a 6-item scale for the assessment of depersonalization (DP) and derealization (DR). The validity of this scale (NI DRP) was examined in comparison to the German version of the Dissociative Experiences Scale (FDS) and the Cambridge Depersonalization Scale (CDS). The sample consists of 144 psychotherapy patients, of whom showed on the basis of a structured clinical interview n = 51 none, n = 45 mild, n = 28 moderate and n = 20 severe DP-DR. The areas under the curve and the rate of misclassification did not differ for NI-DRP and the scales of the FDS. The CDS showed the lowest rate of misclassifications. PMID- 16342025 TI - [Patient and therapist satisfaction in outpatient psychotherapy]. AB - Increasing attention is devoted to the patient perspective in mental health services research. Consequently, patient satisfaction has taken a prominent role and is frequently applied as an indicator of treatment quality and even outcome. However, the suitability of patient satisfaction for assessing quality of mental health services is doubtful, and there is a lack of longitudinal data especially for outpatient psychotherapy. By using data of the TRANS-OP study, we examine the course of patient satisfaction in outpatient psychotherapy over time and its predictors such as symptom change and duration of treatment. Furthermore, the relation between treatment satisfaction ratings of patients and therapists will be scrutinized. This study was conducted by the Center for Psychotherapy Research in Stuttgart (now Heidelberg) in cooperation with the Deutsche Krankenversicherung. N = 714 participants who applied for reimbursement of outpatient psychotherapy were consecutively recruited and asked to provide data at five (therapists two) measurement points during a two-year period. In line with previous evidence, high rates of patient satisfaction have been found, and also therapists appraised favorably the quality of their treatments. Course of patient satisfaction was found to be markedly stable over time. While some variables related to initial patient satisfaction, its course could hardly be predicted. Treatment outcome and session number moderately affected satisfaction at treatment termination. Concordance between patients' and therapists' satisfaction ratings was poor. Implications of these findings for research and practice are discussed. PMID- 16342026 TI - [Brief case report. Duloxetine in Cotard syndrome]. PMID- 16342028 TI - [Publication references. On post-traumatic stress disorder]. PMID- 16342029 TI - [Tabea Apfel: are psychiatric patients referred too quickly for pensions?]. PMID- 16342030 TI - Communal use of integumental wounds in honey bee (Apis mellifera) pupae multiply infested by the ectoparasitic mite Varroa destructor. AB - The ectoparasitic bee mite, Varroa destructor, is highly adapted to its natural and adopted honey bee hosts, Apis cerana and Apis mellifera. Adult females perforate the integument of bee pupae in such a way that they and their progeny can feed. We examined the wounds that founder females made, and usually found one, and rarely up to three, integumental wounds on pupae of A. mellifera multiply infested by V. destructor. The punctures were mainly on the 2nd abdominal sternite of the host. These perforations are used repeatedly as feeding sites by these hemolymph-sucking mites and by their progeny. The diameter of the wounds increased during pupal development. In brood cells containing 4-5 invading female mites and their progeny, healing of the wound is delayed, normally occurring just before the imaginal moult of the bee pupa. These wounds are subject to microbial infections, and they are relevant to the evolution of behavioral traits in these parasitic mites and their relations to host bees. PMID- 16342031 TI - Morphology and protein patterns of honey bee drone accessory glands. AB - We used light and transmission electron microscopy to examine the morphology of the accessory glands of immature and mature adult males of Apis mellifera L. We also made an electrophoretic analysis of the protein content of the mature gland. The glands of the immature male actively secrete a mucous substance that can be seen in the lumen of the gland of the mature male. This secretion stains with mercury bromophenol blue and with periodic acid-Schiff reaction, which stain glyconjugates. The protein content was higher in the lumen secretion than in the gland wall extracts. The electrophoresis patterns of the wall extracts were different from those of the secretion found in the gland lumen. PMID- 16342033 TI - HIV TAT variants differentially influence the production of glucocerebrosidase in Sf9 cells. AB - Gaucher disease, the most common lysosomal storage disorder, is currently treated with enzyme replacement therapy. This approach, however, is ineffective in altering the progression of neurodegeneration in type 2 and type 3 patients due to the difficulty of transferring the recombinant enzyme across the blood-brain barrier. Human immunodeficiency virus type 1 trans-activating transcriptional activator protein (HIV TAT) contains a protein transduction domain that can be added to a fusion protein partner to allow for transport of the partner across membranes. Consequently, we examined the creation, production, and secretion of fusion constructs containing glucocerebrosidase and either wild-type TAT or modified TAT in Sf9 cells. All three constructs exhibited successful expression, with wild-type TAT chimeras showing lower levels of expression than modified TAT chimeras. PMID- 16342032 TI - Genetic and non-genetic effects on productive and reproductive traits of cows in dual-purpose herds in southeastern Mexico. AB - Data on productive and reproductive performance of cows in dual-purpose herds were analyzed to determine the effect of some environmental and genetic factors on saleable milk yield (SMY), lactation length (LL), daily saleable milk per lactation (DMYL), calving interval (CI), and daily saleable milk per calving interval (MYCI) in dual-purpose herds in Yucatan, Mexico. Repeatabilities (re) for these traits were also estimated. Data were obtained from monthly visits to 162 herds from 1996 to 2000. The fixed factors studied were: region (central, eastern and southern), parity number (1 to 6), calving year (1996 to 2000) and calving season (dry, rainy and windy and rainy), genetic group (low- (< 0.50%), medium- (50%) and high- (> 50%) grade cows with European genes). About 2700 to 7700 cows were evaluated for each trait. All factors had significant effects (P < 0.05) on the traits except for region on CI and calving season on DMYL. The overall means for SMY, LL, DMYL, CI, and MYCI were: 1322.3 +/- 80.5 kg, 224.8 +/- 1.3 days, 5.8 +/- 0.1 kg, 555.1 +/- 16.5 days, and 3.0 +/- 0.1 kg, respectively. The re values for SMY, LL, DMYL, CI, and MYCI traits were: 0.19 +/- 0.03, 0.08 +/ 0.04, 0.16 +/- 0.04, 0.00 +/- 0.08, and 0.08 +/- 0.07, respectively. First parity cows had lower SMY, shorter LL, longer CI, and lower MYCI means than cows with more than one parity. Medium grade cows produced more SMY, DMYL and MYCI and had shorter CI than low- and high-grade cows. Therefore, under Yucatan conditions medium-grade cows should be exploited, and more attention should be given to first parity cows in order to improve the productivity in the herd. The relatively high re estimates for SMY and DMYL can be used to calculate most probable producing abilities, in order to identify which cows should be culled. PMID- 16342034 TI - Genetic characterization of European-Zebu composite bovine using RFLP markers. AB - A population of 370 European-Zebu composite beef heifers, consisting of six different breed compositions (A-F), were characterized genetically, using RFLP markers of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) genes. Our objectives were to genetically characterize this population and to determine the structure and the genetic variability of this hybrid herd. The genotypes were determined through PCR, followed by digestion with restriction endonucleases. The PCR-RFLP analysis made it possible to identify the LHR and FSHR genotypes, as well as to characterize the degree of heterozygosis, which was high for all of the breed compositions, for both loci, except for two combinations for LHR (B and C). The observed heterozygosity (Ho) was lower than the expected heterozygosity (He) for compositions C (for LHR) and A and D (for FSHR); however, for the population as a whole, Ho was above He (with a mean of 57 versus 46%, respectively), reflecting the elevated genetic variability in this population and also the informative value of the RFLP markers, which could be useful for population genetic characterization studies. The analysis of the degree of genetic structure of this population, estimated by the Nei's statistic, for both loci, indicated an elevated total genetic diversity (HT = 47%), with most of this variability being due to intrapopulational diversity (HS = 46%), with a low degree of genetic differentiation among the six breed compositions (GST = 1.2%). The estimates generated by the Wright's F statistic indicated a non-endogamic population, with excess heterozygotes (FIT = 0.22), which was also observed at the intrapopulational level (FIS = -0.23). The results gave evidence that the genetic selection applied to this European-Zebu composite population did not affect the expected high genetic variability for this type of crossbreeding, which makes it possible to use these animals to obtain economically valuable productive and reproductive traits. PMID- 16342035 TI - An integrated model for cellular analysis. AB - We present the MOlecular NETwork (MONET) ontology as a model to integrate data from different networks that govern cell function. To achieve this, different existing ontologies were analyzed and an integrated ontology was built in a way to make it possible to share and reuse knowledge, support interoperability between systems, and also allow the formulation of hypotheses through inferences. By studying the cell as an entity of a myriad of elements and networks of interactions, we aim to offer a means to understand the large-scale characteristics responsible for the behavior of the cell and to enable new biological insights. PMID- 16342036 TI - Recent advances in gene expression data clustering: a case study with comparative results. AB - Several advanced techniques have been proposed for data clustering and many of them have been applied to gene expression data, with partial success. The high dimensionality and the multitude of admissible perspectives for data analysis of gene expression require additional computational resources, such as hierarchical structures and dynamic allocation of resources. We present an immune-inspired hierarchical clustering device, called hierarchical artificial immune network (HaiNet), especially devoted to the analysis of gene expression data. This technique was applied to a newly generated data set, involving maize plants exposed to different aluminum concentrations. The performance of the algorithm was compared with that of a self-organizing map, which is commonly adopted to deal with gene expression data sets. More consistent and informative results were obtained with HaiNet. PMID- 16342037 TI - A multi-neighbor-joining approach for phylogenetic tree reconstruction and visualization. AB - The computationally challenging problem of reconstructing the phylogeny of a set of contemporary data, such as DNA sequences or morphological attributes, was treated by an extended version of the neighbor-joining (NJ) algorithm. The original NJ algorithm provides a single-tree topology, after a cascade of greedy pairing decisions that tries to simultaneously optimize the minimum evolution and the least squares criteria. Given that some sub-trees are more stable than others, and that the minimum evolution tree may not be achieved by the original NJ algorithm, we propose a multi-neighbor-joining (MNJ) algorithm capable of performing multiple pairing decisions at each level of the tree reconstruction, keeping various partial solutions along the recursive execution of the NJ algorithm. The main advantages of the new reconstruction procedure are: 1) as is the case for the original NJ algorithm, the MNJ algorithm is still a low-cost reconstruction method; 2) a further investigation of the alternative topologies may reveal stable and unstable sub-trees; 3) the chance of achieving the minimum evolution tree is greater; 4) tree topologies with very similar performances will be simultaneously presented at the output. When there are multiple unrooted tree topologies to be compared, a visualization tool is also proposed, using a radial layout to uniformly distribute the branches with the help of well-known metaheuristics used in computer science. PMID- 16342038 TI - A framework based on Web service orchestration for bioinformatics workflow management. AB - Bioinformatics activities are growing all over the world, with proliferation of data and tools. This brings new challenges: how to understand and organize these resources and how to provide interoperability among tools to achieve a given goal. We defined and implemented a framework to help meet some of these challenges. Four issues were considered: the use of Web services as a basic unit, the notion of a Semantic Web to improve interoperability at the syntactic and semantic levels, and the use of scientific workflows to coordinate services to be executed, including their interdependencies and service orchestration. PMID- 16342039 TI - Reconfigurable systems for sequence alignment and for general dynamic programming. AB - Reconfigurable systolic arrays can be adapted to efficiently resolve a wide spectrum of computational problems; parallelism is naturally explored in systolic arrays and reconfigurability allows for redefinition of the interconnections and operations even during run time (dynamically). We present a reconfigurable systolic architecture that can be applied for the efficient treatment of several dynamic programming methods for resolving well-known problems, such as global and local sequence alignment, approximate string matching and longest common subsequence. The dynamicity of the reconfigurability was found to be useful for practical applications in the construction of sequence alignments. A VHDL (VHSIC hardware description language) version of this new architecture was implemented on an APEX FPGA (Field programmable gate array). It would be several magnitudes faster than the software algorithm alternatives. PMID- 16342040 TI - Mutual information content of homologous DNA sequences. AB - The necessary information to reproduce and keep an organism is codified in acid nucleic molecules. Deepening the knowledge about how the information is stored in these bio-sequences can lead to more efficient methods of comparing genomic sequences. In the present study, we analyzed the quantity of information contained in a DNA sequence that can be useful to identify sequences homologous to it. To reach it, we used signal processing techniques, specially spectral analysis and information theory. PMID- 16342041 TI - An I/O device driver for bioinformatics tools: the case for BLAST. AB - There are many bioinformatics tools that deal with input/output (I/O) issues by using filing systems from the most common operating systems, such as Linux or MS Windows. However, as data volumes increase, there is a need for more efficient disk access, ad hoc memory management and specific page-replacement policies. We propose a device driver that can be used by multiple applications. It keeps the application code unchanged, providing a non-intrusive and flexible strategy for I/O calls that may be adopted in a straightforward manner. With our approach, database developers can define their own I/O management strategies. We used our device driver to manage Basic Local Alignment Search Tool (BLAST) I/O calls. Based on preliminary experimental results with National Center for Biotechnology Information (NCBI) BLAST, this approach can provide database management systems like data management features, which may be used for BLAST and many other computational biology applications. PMID- 16342042 TI - An agent-based system for re-annotation of genomes. AB - Genome annotation projects can produce incorrect results if they are based on obsolete data or inappropriate models. We have developed an automatic re annotation system that uses agents to perform repetitive tasks and reports the results to the user. These tasks involve BLAST searches on biological databases (GenBank) and the use of detection tools (Genemark and Glimmer) to identify new open reading frames. Several agents execute these tools and combine their results to produce a list of open reading frames that is sent back to the user. Our goal was to reduce the manual work, executing most tasks automatically by computational tools. A prototype was implemented and validated using Mycoplasma pneumoniae and Haemophilus influenzae original annotated genomes. The results reported by the system identify most of new features present in the re-annotated versions of these genomes. PMID- 16342043 TI - Reconstruction of phylogenetic trees using the ant colony optimization paradigm. AB - We developed a new approach for the reconstruction of phylogenetic trees using ant colony optimization metaheuristics. A tree is constructed using a fully connected graph and the problem is approached similarly to the well-known traveling salesman problem. This methodology was used to develop an algorithm for constructing a phylogenetic tree using a pheromone matrix. Two data sets were tested with the algorithm: complete mitochondrial genomes from mammals and DNA sequences of the p53 gene from several eutherians. This new methodology was found to be superior to other well-known softwares, at least for this data set. These results are very promising and suggest more efforts for further developments. PMID- 16342044 TI - A distributed computation of Interpro Pfam, PROSITE and ProDom for protein annotation. AB - Interpro is a widely used tool for protein annotation in genome sequencing projects, demanding a large amount of computation and representing a huge time consuming step. We present a strategy to execute programs using databases Pfam, PROSITE and ProDom of Interpro in a distributed environment using a Java-based messaging system. We developed a two-layer scheduling architecture of the distributed infrastructure. Then, we made experiments and analyzed the results. Our distributed system gave much better results than Interpro Pfam, PROSITE and ProDom running in a centralized platform. This approach seems to be appropriate and promising for highly demanding computational tools used for biological applications. PMID- 16342045 TI - Gene selection based on multi-class support vector machines and genetic algorithms. AB - Microarrays are a new technology that allows biologists to better understand the interactions between diverse pathologic state at the gene level. However, the amount of data generated by these tools becomes problematic, even though data are supposed to be automatically analyzed (e.g., for diagnostic purposes). The issue becomes more complex when the expression data involve multiple states. We present a novel approach to the gene selection problem in multi-class gene expression based cancer classification, which combines support vector machines and genetic algorithms. This new method is able to select small subsets and still improve the classification accuracy. PMID- 16342046 TI - Support vector machines for novel class detection in Bioinformatics. AB - Novelty detection techniques might be a promising way of dealing with high dimensional classification problems in Bioinformatics. We present preliminary results of the use of a one-class support vector machine approach to detect novel classes in two Bioinformatics databases. The results are compatible with theory and inspire further investigation. PMID- 16342047 TI - [Factors predicting outcome in renal transplantation]. AB - Organ shortage and necessity of good allocation to improve graft outcomes are well documented by epidemiological data. The authors analyze the main factors influencing long-term graft outcome concerning donor, peritransplant period and recipient. Donor age, marginal donors and new knowledges on donor brain death are the most relevant donor related factors. Cold ischemia is still a relevant peritransplant factor. Its influence on dentritic cell activation has been recently discovered. Regarding histocompatibility the six antigen match condition undoubtedly is associated to a better outcome. Most relevant recipient factors are: Preemptive transplant from living and deceased donors, whenever possible. With improvements in long-term kidney graft survival,the recurrence of original kidney disease is becoming an important cause of late graft failure. Recently several biological markers are emerging as graft outcome predictors. Recent findings indicate that certain cytokines or special lymphocyte subsets (alloreactive T memory cells) in the recipient could be powerful markers in the next future. PMID- 16342048 TI - [Maintenance and monitoring of water treatment system]. AB - Water treatment systems must be submitted to maintenance, disinfections and monitoring periodically. The aim of this review is to analyze how these processes must complement each other in order to preserve the efficiency of the system and optimize the dialysis fluid quality. The correct working of the preparatory process (pre-treatment) and the final phase of depuration (reverse osmosis) of the system need a periodic preventive maintenance and the regular substitution of worn or exhausted components (i.e. the salt of softeners' brine tank, cartridge filters, activated carbon of carbon tanks) by a competent and trained staff. The membranes of reverse osmosis and the water distribution system, including dialysis machine connections, should be submitted to dis-infections at least monthly. For this purpose it is possible to use chemical and physical agents according to manufacturer' recommendations. Each dialysis unit should predispose a monitoring program designed to check the effectiveness of technical working, maintenance and disinfections and the achievement of chemical and microbiological standards taken as a reference. Generally, the correct composition of purified water is monitored by continuous measuring of conductivity, controlling bacteriological cultures and endotoxin levels (monthly) and checking water contaminants (every 6-12 months). During pre-treatment, water hardness (after softeners) and total chlorine (after chlorine tank) should be checked periodically. Recently the Italian Society of Nephrology has developed clinical guidelines for water and dialysis solutions aimed at suggesting rational procedures for production and monitoring of dialysis fluids. It is hopeful that the application of these guidelines will lead to a positive cultural change and to an improvement in dialysis fluid quality. PMID- 16342049 TI - [HIV-1 and renal cells: pathogenesis of HIV-associated nephropathy]. AB - Kidney is one of the organs, as haematopoietic tissue and central nervous system, representing a reservoir of HIV-1, where the virus can exert a direct pathogenic activity. HIV-associated nephropathy (HIVAN) is the prominent illness among the numerous renal injuries occurring in HIV-1 infection. It is characterized by heavy proteinuria and rapid progression to end stage renal disease. Histopathologically, HIVAN is a collapsing form of focal segmental glomerulosclerosis with podocyte hyperplasia and dedifferentiation, associated with severe tubulopathy which is characterized by tubular apoptosis, microcysts, and interstitial fibrosis. Several clinical and experimental data point to a direct role of HIV-1 in kidney damage. In renal biopsies of HIVAN cases viral transcripts have been found in glomerular and tubular epithelial cells. Transgenic mice expressing replication-defective HIV proviral constructs develop a renal disease similar to HIVAN both from the histopathological and clinical point of view. In vitro studies using cultures of human renal cells have shown that HIV-1 can induce in glomerular and tubular cells distinct pathogenic effects, which mimic the pathological features of HIVAN in vivo. A large body of evidence suggests that an abnormal response of podocytes to HIV-1 infection and/or HIV-1 proteins is the key event in HIVAN pathogenesis. Since the highly active antiretroviral therapy has demonstrated scant beneficial effects on the development of HIVAN, the elucidation of the pathogenic mechanisms activated by HIV-1 in the renal cells might allow designing specific therapeutic strategies. PMID- 16342050 TI - [Protein metabolism in renal failure]. AB - Patients with chronic renal failure are often characterized by clinical and laboratory signs of protein and calories malnutrition, which is associated with a significant increase in the total morbidity and mortality risk. The loss of muscle mass is often multifactorial. If the patient is consuming a low protein diet the risk of protein calories malnutrition is increased. Among the various factors that can affect protein metabolism the role of calories and protein intake as well the influences of concomitant conditions as increased leptin levels and uncontrolled metabolic acidosis are evaluated. Hemodialysis treatment induces a transient increase in proteolysis which is associated with a significant loss of amino acid in the dialysis fluid. Subclinical inflammation contributes to malnutrition increasing both muscle proteolysis and anorexia. Albumin levels are often reduced in dialysis patients, this decline is associated with an increase in morbidity and mortality. Various factors as protein intake, acidosis and inflammation can affect albumin levels via a reduction in hepatic albumin synthesis. PMID- 16342051 TI - [Hyperhomocysteinemia and progression of renal disease]. AB - The relationship between hyperhomocysteinemia and cardiovascular damage is well known, whereas the role of this alteration in renal disease progression has been scarcely studied. Experimental studies demonstrated that exposure to high levels of homocysteinemia causes glomerular and interstitial damage which is remarcably proportional to the serum concentration of this aminoacid. Until now the renal effects of hyperhomocysteinemia in man has been investigated only in observational studies. The Hoorn study, a prospective study in a Dutch population, showed that the plasma homocysteine is a strong predictor of proteinuria in diabetic and non diabetic subjects. Findings in this study were recently confirmed in a cohort study in 7500 Japanese because plasma homocysteine predicted the onset of renal failure in this population. NO-dependent endothelial dysfunction triggered by homocysteine via reduction of the activity of the enzyme that metabolizes Asimmetric Dymethilarginine (dmethylarginine dymethilaminohydrolase) is a likely mechanism whereby hyperomocysteinemia causes cardiovascular and renal damage as well. PMID- 16342052 TI - [Historical Archives of Italian Nephrology. Gabriele Monasterio (1903-1972) and the School of Pisa]. AB - Gabriele Monasterio (1903-1972), a clinician with numerous medical interests, contributed remarkably to the development of the Italian nephrology of his time. His main scientific contributions were: - Investigation of the causes of normoglycaemic glycosuria. Of this condition he collected, from 1939 on, a large series of patients whom he also investigated - for the first time in the world - with renal biopsy and - thanks to a collaboration with Jean Oliver - the microdissection of the nephron. - Introduction, in the early 1960s, of a low protein diet for the patients with chronic renal failure. This, thanks to the stud-ies published by Monasterio co-workers Quirino Maggiore and Sergio Giovannetti, was largely used in Italy and abroad. - Classification of the nephropathies. This was described in the monograph Le Nefropatie Mediche (1954 and 1970), which had a large impact and today represents a classic of the Italian nephrological literature. Monasterio served remarkably the Italian Society of Nephrology: in 1957 as a member of the founding committee; from 1959 to 1962 and from 1964 to 1968 as President; in 1958, 1962, and 1964 as organizer of the National Congress; from 1962 to his death as the Editor-in-Chief of the official journal of the Society, Minerva Nefrologica. Finally, Monasterio was also a member of the first council of the International Society of Nephrology. PMID- 16342053 TI - [From the Mailing List SIN: professional risks for the nephrologists: clinical risk and risk management]. AB - An extensive debate on bacteraemias due to aquatic bacteria has recently developed in the Mailing List SIN. This topic was analyzed in two previous issues where the epidemics observed in two Dialysis Centers have been presented and discussed. The two experiences had in common the difficulty in identifying the source of contamination in industrial products commercialized as ''sterile''. In one case, the source of the epidemic could not be identified, in the other one it was discovered only incidentally thanks to a notice of the supplier company of the contaminated material. This underlines the importance of defining specific operating protocols in case of hospital epidemics and stresses the issues related to the professional risk and the potential legal implications. PMID- 16342054 TI - [Refractory hypertention in a female patient with renal failure]. AB - We report one sixty-seven years-old female who presented with hypertension refractory to antihypertensive drugs. She had an elevated BP for approximately 15 years. In the last 8-10 months her hypertension had become difficult to control. Her BP ranged between 180/100 mmHg and 220/1220 mmHg on atenolol 100 mg once daily, methyldopa 500 mg three times daily, furosemide 25 mg twice daily, doxazosine 4 mg twice daily. When she was referred to our unit serum creatinine was 2.3 mg/dL and she had a mild proteinuria (70 mg/dL) without microematuria. Ultrasonography showed a left kidney size in the low-normal range (LD 11 cm) and a small right kidney (LD 9 cm). Renal angiography showed a severe, ostial stenosis of the left renal artery and a total thrombosis of the right renal artery with a blood supply to the right kidney provided by collateral channels. An ACE-I was added to the therapy but a sharp increase in serum creatinina (up to 6.4 mg/dL) prompted us to withdraw the drug. She underwent a renal angioplasty on the left side and a Palmaz stent was placed. The control angiography showed a good anatomical result. Three months after the manoeuvre the patient was again referred to our unit with headache, nausea vomiting and hyper-tension refractory to amlodipine 10 mg/day, doxazosine 4 mg twice a a day, atenolol 50 mg/day, furosemide 50 mg/day. A doppler ultrasonography and a magnetic resonance angiogram showed no restenosis on the treated artery. An ACE-I was again administered and BP on this drug was 145/90 mmHg after one month and 130/85 after three months. Headache, nausea and vomiting disappeared. Serum creatinina kept unchanged (2.2 mg/dL). Comment. In this case the benefit of angioplasty on blood pressure control was indirect. Apparently the manoeuvre showed no effect on blood pressure, but the angioplasty allowed us to use of an ACE-Inhibitor, without any negative effect on renal function, and thus to adequately control blood pressure. PMID- 16342055 TI - Velocity-selective RF pulses in MRI. AB - A family of velocity-selective pulses consisting of a series of RF hard pulses followed by bipolar gradients was designed. The succession of required pulses was deduced using a k-space approach within a small tip-angle approximation. Fourier transform of the desired velocity excitation determined the flip-angle series, and the corresponding position in the generalized k-space identified the bipolar gradient first moments. Spins from any velocity class can be selected. To illustrate this approach we designed and experimentally tested a velocity-slice selection that is analogous to standard spatial-slice selection but involves excitation of spins moving at a chosen velocity (velocity-slice center) and within a given interval (velocity-slice thickness). The assumed approximation does not limit the design to small angles, because velocity selection still holds for angles up to 90 degrees. Velocity slices were experimentally selected, centered on velocities ranging from -1 m s(-1) to 1 m s(-1) with a velocity-slice thickness of 0.4 m s(-1). The experimental velocity-slice profile was assessed and the flow was quantified. PMID- 16342056 TI - Presynaptic GABA(B) receptors on glutamatergic terminals of CA1 pyramidal cells decrease in efficacy after partial hippocampal kindling. AB - We tested the hypothesis that presynaptic GABA(B) receptors on glutamatergic terminals (GABA(B) heterosynaptic receptors) decreased in efficacy after partial hippocampal kindling. Rats were implanted with chronically indwelling electrodes and 15 hippocampal afterdischarges were evoked by high-frequency electrical stimulation of hippocampal CA1. Control rats were implanted with electrodes but not given high-frequency stimulations. One to 21 days after the last afterdischarge, excitatory postsynaptic potentials (EPSPs) were recorded in CA1 of hippocampal slices in vitro, following stimulation of the stratum radiatum. Field EPSPs (fEPSPs) were recorded in CA1 stratum radiatum and intracellular EPSPs (iEPSPs) were recorded from CA1 pyramidal cells. GABA(B) receptor agonist +/- baclofen (10 microM) in the bath suppressed the fEPSPs significantly more in control than kindled rats, at 1 or 21 days after kindling. Similarly, baclofen (10 microM) suppressed iEPSPs more in the control than the kindled group of neurons recorded at 1 day after kindling. Suppression of the fEPSPs by 1 microM N(6)-cyclopentyladenosine, which acted on presynaptic A1 receptors, was not different between kindled and control rats. Activation of the GABA(B) heteroreceptors by a conditioning burst stimulation of CA3 afferents suppressed the iEPSPs evoked by a test pulse. The suppression of the iEPSPs at 250-500 ms condition-test interval was larger in control than kindled groups of neurons. It was concluded that the efficacy of presynaptic GABA(B) receptors on the glutamatergic terminals was reduced after partial hippocampal kindling. The reduction in heterosynaptic presynaptic GABA(B) receptor efficacy will increase glutamate release and seizure susceptibility, particularly during repeated neural activity. PMID- 16342057 TI - Differences in the allosteric modulation by phenytoin of the binding properties of the sigma1 ligands [3H](+)-pentazocine and [3H]NE-100. AB - The present study evaluated the effects of phenytoin (DPH) on the binding to synaptosomal fraction membranes from guinea pig brain of the prototypic sigma1 (sigma1) receptor agonist [3H](+)-pentazocine and the putative sigma1 antagonist [3H]NE-100. Equilibrium and binding kinetics studies were done. The order of affinity of 12 sigma1 ligands for binding sites labeled with [3H](+)-pentazocine correlated well with their order of affinity for sites labeled with [3H]NE-100, suggesting that both radioligands label the same receptor. Phenytoin increased the binding of [3H](+)-pentazocine, enhancing its affinity (K(D) value) for sigma1 receptors and decreasing its dissociation rate from these receptors. The maximal number of receptors (B(max) value) labeled with [3H](+)-pentazocine was not changed. In contrast, phenytoin decreased the specific binding and maximal number of receptors labeled with [3H]NE-100, and increased its dissociation rate from sigma1 receptors. The affinity of this radioligand for sigma1 receptors was not modified. In conclusion, phenytoin behaved as a positive allosteric modulator on the binding of [3H](+)-pentazocine, whereas it negatively modulated the binding of [3H]NE-100. These results add evidence in favor of the use of phenytoin in vitro to distinguish between agonists and antagonists of sigma1 receptors. PMID- 16342058 TI - Reversible changes in hippocampal CA1 synapses associated with water maze training in rats. AB - Long-term memories seem to require protein synthesis to be established. This process can be related with synaptogenesis resulting in changes in the form or even in the number or proportion of synaptic contacts. Results from behavioral studies assessing quantitative changes associated with different learning tasks are controversial. The aim of our work was to assess whether the number of CA1 hippocampal synaptic contacts can be modified after training in different tasks in the Morris water maze (MWM). We found transient changes in the synaptic density of the symmetric synapses associated with place learning. A reduced synaptic density of the symmetric synapses in the stratum radiatum of CA1 was found at 48 h posttraining, returning to control levels 72 h posttraining. The same effect was observed 1 h after training in a nonspatial task. Synaptic changes found in the CA1 shortly after water maze training suggest a possible participation of the hippocampus in the acquisition of nonspatial tasks together with a role in the short-term consolidation of spatial memory. As no changes were found in the total number of synapses counted, it is likely that subtle changes in synaptic efficacy than new synapse generation may be sufficient to support the acquisition and maintenance of new memories. PMID- 16342059 TI - Lesion of the lateral entorhinal cortex amplifies odor-induced expression of c fos, junB, and zif 268 mRNA in rat brain. AB - Paradoxical facilitation of olfactory learning following entorhinal cortex (EC) lesion has been described, which may result from widespread functional alterations taking place within the olfactory system. To test this hypothesis, expression of the immediate early genes c-fos, junB, and zif 268 was studied in response to an olfactory stimulation in several brain areas in control and in EC lesioned rats. Olfactory stimulation in control rats induced the expression of the three genes in the granular/mitral and glomerular layers of the olfactory bulb, as well as c-fos and junB expression in the piriform cortex. However EC lesion was devoid of effects in nonstimulated animals; it significantly amplified the odor-induced expression of the three genes in these areas, as well as in the amygdala, hippocampus, and parietal-temporal cortices. The data suggest that EC lesion modifies the neural processing of odor by suppressing an inhibitory influence on brain areas connected to this cortex. PMID- 16342060 TI - A robust ratio metric method for analysis of Zap-70 expression in chronic lymphocytic leukemia (CLL). AB - Since Zap-70 expression in chronic lymphocytic leukemia (CLL) cells correlates with a lack of somatic mutation of the immunoglobulin variable heavy chain (IgVH) genes, it has been proposed as a surrogate marker for disease prognosis. However, published studies of Zap-70 expression have used different commercial antibodies and analytic strategies. This study was undertaken to determine if any strategy was broadly applicable in a clinical flow cytometry laboratory. Expression of Zap 70 was determined in 37 CLL patients using four different commercial antibodies. T, NK, and CLL cells were identified by immunophenotyping along with Zap-70 expression. Data was analyzed in terms of both percent of CLL cells expressing Zap-70 and the ratio of Zap-70 expression in CLL cells compared to that in T + NK cells. Three Zap-70 antibodies showed wide ranges of Zap-70 expression as a percentage of tumor cells, while a fourth gave consistently elevated results. Comparing the percent Zap-70 expression with any two antibodies gave poor correlations (r(2) = 0.45-0.63). Our results indicated that the previous analytical strategies were not reproducible. A ratio metric is proposed, which gave better correlations (r(2) as high as 0.95) and would allow separation of CLL patients with elevated or decreased Zap-70 expression. PMID- 16342061 TI - An overview on the development of a bio-artificial pancreas as a treatment of insulin-dependent diabetes mellitus. AB - This paper presents the concept and most of the research undertaken all over the world for the development of a bio-artificial pancreas (BAP) device over the last 30 years. The devices studied, meant to mimic the insulin secretion of the natural organ, were diverse and have been reviewed. Allogeneic or xenogeneic cells or cell clusters have been separated from the host's immune system by synthetic biocompatible semipermeable membranes to prevent the need, of the host, for immune-suppressing regimens. The biocompatible polymer used as a barrier and its intrinsic characteristics, the cell immobilization or suspension media, the existence or not of co-immobilized molecules or cells, the number of devices used and the implantation site, were addressed. PMID- 16342062 TI - Flow-cytometric analysis of reactive oxygen species in peripheral blood mononuclear cells of patients with thyroid dysfunction. AB - BACKGROUND: Thyroid hormones are major regulators of energy metabolism and increased levels of the hormones (hyperthyroidism) results in an increase in the metabolic rate. Thyroid dysfunction causing alteration in hormone secretion leads to perturbations in the metabolic status. The hypermetabolic state may cause increased generation of reactive oxygen species (ROS), leading to oxidative stress in these patients. This study was carried out to verify our proposition by measuring the ROS in the terminally differentiated cells like the peripheral blood mononuclear cells of the patients. METHODS: Flow-cytometric analysis of the ROS was carried out using 2',7' dichlorofluorescein diacetate in the isolated peripheral blood mononuclear cells of the subjects. RESULTS: ROS generation was found to be 3-folds higher in hyperthyroids as compared with euthyroids and hypothyroids and this was not found to be gender specific. CONCLUSIONS: Hyperthyroidism results in ROS generation in patients, which can be detected flow cytometrically in the peripheral blood mononuclear cells. Hence, this could complement the other thyroid function tests facilitating the diagnosis and design of appropriate therapy. PMID- 16342063 TI - Wilms tumour experience in a South African centre. AB - BACKGROUND: In Africa, Wilms tumour frequently presents with advanced disease. This study reports our results over 25 years using the National Wilms Tumour Study Group (NWTSG) approach of primary surgery followed by chemotherapy. PROCEDURE: A retrospective analysis was performed on all patients diagnosed with unilateral Wilms tumour between January 1979 and December 2003. RESULTS: The records of 188 children with newly diagnosed Wilms tumour were examined. Of these 57 (30.3%) were stage I, 33 (17.6%) were stage II, 60 (31.9%) were stage III and 38 (20.2%) were stage IV. Twenty-four patients (12.8%) had unfavourable histology (UFH). The estimated 5-year overall survival (OS) for the whole group was 80.5% (84.8% for favourable histology (FH) and 51.6% for UFH). Among those with FH, estimated 5-year OS was 94.4% for stage I, 96.2% for stage II, 84.9% for stage III and 54.2% for stage IV. There was no difference in OS between those FH stage III tumours that were operable and those deemed inoperable. Intra-operative spillage was uncommon (8%), and did not increase local relapse rate. CONCLUSIONS: National Wilms Tumour Study Group protocols employed in an African setting with highly competent and experienced surgical care, produced results for non metastatic FH Wilms tumour comparable to those of the NWTSG. PMID- 16342064 TI - Impact of evidence-based interdisciplinary guidelines on testis cancer management. AB - BACKGROUND: In 1999, interdisciplinary evidence-based guidelines were elaborated for treatment of germ cell tumors in Germany. The aims of the current study were to analyze failures in diagnosis and therapy and to demonstrate the influence of guidelines on individual therapeutic approaches and clinical outcome. Therefore, patient collectives treated before the introduction of guidelines (Group A, 1990 1999, n = 234) and those thereafter (Group B, 2000-2002, n = 84) were compared for recurrence and survival. METHODS: In both groups, medical and/or surgical treatment and clinical outcome were evaluated for therapeutic mistakes and violations of guidelines. These were analyzed for their clinical consequences. RESULTS: There was no significant difference between groups concerning median age of patients or clinical stage before therapy. Altogether, 27.8% and 8.3% of all patients in Group A and B, respectively, displayed therapeutic mistakes (P < 0.005); 63% of these patients in Group A and 100% of these patients in Group B received an overtreatment. In Group A, 19/234 (8.1%) patients relapsed and 53% of these patients had been treated insufficiently (P < 0.005). Advanced disease caused the death of 3/234 patients in this study. As of this writing, only 3 of 84 (3.6%) patients in Group B have relapsed, and no patient has died because of tumor or consecutive treatment. CONCLUSIONS: The integration of interdisciplinary evidence-based guidelines for treatment of testicular germ cell tumors has led to significant reduction of both overtreatment and treatment failure and/or relapse that were due to inappropriate primary therapy. Evidence-based guidelines should serve as internal quality controls in all institutions treating patients with testicular germ cell tumors. PMID- 16342066 TI - Oral mucositis in patients undergoing radiation treatment for head and neck carcinoma. AB - BACKGROUND: The current study was conducted to characterize the risks and clinical consequences of oral mucositis (OM) in patients with head and neck carcinoma (HNC) who are receiving radiation therapy. METHODS: Data regarding 450 HNC patients who had received radiation therapy were collected via chart review from 154 U.S. medical and radiation oncologists. Information obtained included patient characteristics, treatments received, highest recorded grade of OM during radiation therapy (none, mild, moderate, or severe), and outcomes potentially associated with mucosal injury. RESULTS: The mean age (+/- standard deviation [SD]) of the study subjects was 61.3 years (12.3 yrs); the majority of patients (80%) were men. Primary tumor locations included the oropharynx (26.4%), larynx (26.4%), oral cavity including the lip (24.4%), hypopharynx (13.6%), and nasopharynx (9.1%). The majority of tumors were new and were classified as AJCC Stages III or IV. The majority of patients (83%) received standard radiation therapy; the mean (+/- SD) cumulative dose was 6285 centigrays (cGy) (+/- 1158 cGy). Approximately 33% of the patients received concomitant chemotherapy. The majority of patients (83%) developed OM; 29% developed severe OM. Patients with severe OM were more likely to have nasopharyngeal or oropharyngeal tumors (adjusted odds ratio [OR] of 10.1 [95% confidence interval (95% CI), 2.1-49.9] and 6.9 [95% CI, 2.4-19.7], respectively), and to have received cumulative radiation doses > 5000 cGy (OR of 10.4; 95% CI, 2.9-37.1) and concomitant chemotherapy (OR of 3.3; 95% CI, 1.4-8.0). Patients with OM had more unplanned breaks in radiation therapy (OR of 3.8; 95% CI, 1.7-8.5) and hospital admissions (OR of 3.5; 95% CI, 1.3-9.5). CONCLUSIONS: HNC patients with nasopharyngeal or oropharyngeal tumors, and those who receive cumulative radiation doses > 5000 cGy or concomitant chemotherapy, are more likely to develop OM. Patients with OM are at a higher risk of unplanned breaks in radiation therapy and hospitalization. PMID- 16342065 TI - The combination of gemcitabine and carboplatin as first-line treatment in patients with advanced urothelial carcinoma. A Phase II study of the Hellenic Cooperative Oncology Group. AB - BACKGROUND: The toxicity of platinum-based combinations represents a common problem for patients with advanced urothelial carcinoma. The authors previously reported encouraging efficacy for the combination of carboplatin and gemcitabine in patients considered to be unfit for cisplatin-based treatment. The objective of the current multicenter Phase II study was to evaluate the safety and efficacy of the combination of gemcitabine and carboplatin as first-line treatment in unselected patients with advanced urothelial carcinoma. METHODS: Patients with previously untreated, bidimensionally measurable, inoperable or metastatic urothelial carcinoma were treated with carboplatin, area under the concentration curve of 5 (Day 1) and gemcitabine at a dose of 1000 mg/m(2) (Days 1 and 8), every 21 days for a total of 6 cycles. RESULTS: Sixty patients (49 men and 11 women, with a median age of 69 yrs) were enrolled in the current study. Intent-to treat analysis demonstrated an objective response rate (ORR) of 38.4% (95% confidence interval [95% CI], 26-51.8%) (11.7% complete responses and 26.7% partial responses). The median time to disease progression was 7.6 months (95% CI, 4.5-10.7 mos) and the median overall survival was 16.3 months (95% CI, 12 20.6 mos). The median survival was comparable to that reported for the combination of methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC) according to the Memorial Sloan-Kettering Cancer Center prognostic model for patients with similar baseline prognostic features. Grade 3 or 4 toxicity (according to the National Cancer Institute Common Toxicity Criteria [version 2.0]) included anemia (18%), thrombocytopenia (23%), and neutropenia (52%), with 7 episodes of febrile neutropenia (11%) reported. Nonhematologic toxicity was rare. One toxic death occurred during the study. CONCLUSIONS: The combination of gemcitabine and carboplatin appears to have considerable activity as the first line treatment of unselected patients with advanced urothelial carcinoma with manageable toxicity, and deserves further evaluation in this setting. PMID- 16342067 TI - Association between glutathione S-transferase pi polymorphisms and survival in patients with advanced nonsmall cell lung carcinoma. AB - BACKGROUND: Glutathione S-transferase (GST) pi (GSTP1) is a detoxification enzyme with substrate specificity for both exogenous carcinogens and chemotherapy agents. Genetic polymorphisms of GSTP1 exon 5 (Ile105Val) and exon 6 (Ala114Val) appear to reduce this enzyme's activity. Previously, the authors reported that the exon 6 variant was associated with an increased risk of lung carcinoma, particularly among men, younger patients, and ever smokers. In this study, the authors hypothesized that variant GSTP1 genotype would result in reduced inactivation of chemotherapy agents and improved survival in patients with advanced-stage nonsmall cell lung carcinoma (NSCLC), a population that is likely to receive platinum-based chemotherapy. METHODS: Patients with Stage III and IV NSCLC who were enrolled in a molecular epidemiology study were identified, and a polymerase chain reaction-restriction fragment length polymorphism assay was used to genotype GSTP1 exons 5 and 6 in 424 patients and 425 patients, respectively. RESULTS: Patients who had the exon 6 variant genotype (Ala/Val or Val/Val) had significantly better survival compared with patients who had the wild type genotype (Ala/Ala; P = 0.037), with median survival of 16.1 months and 11.4 months, respectively. Multivariate analysis revealed a reduced adjusted hazard ratio (HR) of death associated with the exon 6 variant genotype of 0.75 (95% confidence interval [95% CI], 0.54-1.05). This protective association was observed in younger patients (younger than age 62 yrs; HR, 0.59; 95% CI, 0.57 0.97) and in males (HR, 0.64; 95% CI, 0.41-0.99). GSTP1 exon 5 genotype was not associated with survival. CONCLUSIONS: GSTP1 exon 6 variant genotypes may be associated with improved survival among patients with Stage III and IV NSCLC. PMID- 16342068 TI - Advantage of dichromats over trichromats in discrimination of color-camouflaged stimuli in nonhuman primates. AB - Due to a middle- to long-wavelength-sensitive (M/LWS) cone opsin polymorphism, there is considerable phenotypic variation in the color vision of New World monkeys. Many females have trichromatic vision, whereas some females and all males have dichromatic vision. The selective pressures that maintain this polymorphism are unclear. In the present study we compared the performance of dichromats and trichromats in a discrimination task. We examined tri- and dichromatic individuals of two species: brown capuchin monkeys (Cebus apella) and long-tailed macaques (Macaca fascicularis). We also examined one protanomalous chimpanzee (Pan troglodytes). The subjects' task was to discriminate a circular pattern from other patterns in which textural elements differed in orientation and thickness from the background. After they were trained with stimuli of a single color, the subjects were presented with color-camouflaged stimuli with a green/red mosaic overlaid onto the pattern. The dichromatic monkeys and the protanomalous chimpanzee selected the correct stimulus under camouflaged conditions at rates significantly above chance levels, while the trichromats did not. These findings demonstrate that dichromatic nonhuman primates possess a superior visual ability to discriminate color-camouflaged stimuli, and that such an ability may confer selective advantages with respect to the detection of cryptic foods and/or predators. PMID- 16342069 TI - Color-vision polymorphism in wild capuchins (Cebus capucinus) and spider monkeys (Ateles geoffroyi) in Costa Rica. AB - New World monkeys are unique in exhibiting a color-vision polymorphism due to an allelic variation of the red-green visual pigment gene. This makes these monkeys excellent subjects for studying the adaptive evolution of the visual system from both molecular and ecological viewpoints. However, the allele frequencies of the pigments within a natural population have not been well investigated. As a first step toward understanding the relationship between vision and behavior, we conducted color-vision typing by analyzing fecal DNA from two wild groups of white-faced capuchin monkeys (Cebus capucinus) and one group of black-handed spider monkeys (Ateles geoffroyi) inhabiting Santa Rosa National Park of Costa Rica. All color-typed monkeys were individually identified. In C. capucinus and A. geoffroyi we found three and two pigment types, respectively, and the spectral mechanism that created one of the two Ateles pigments was found to be novel. In one Cebus group and the Ateles group, all alleles were present, whereas in the other Cebus group only two alleles were found, with one allele predominating. This was likely due to the effect of close inbreeding, indicating that wild populations can exhibit a variety of allele compositions. This result also suggests that the color-vision polymorphism can be easily distorted by natural factors, such as inbreeding, skewing the population structure. PMID- 16342070 TI - Demonstration of a genotype-phenotype correlation in the polymorphic color vision of a non-callitrichine New World monkey, capuchin (Cebus apella). AB - Color-vision polymorphism in New World monkeys occurs because of an allelic polymorphism of the single-copy red-green middle-to-long-wavelength-sensitive (M/LWS) opsin gene on the X chromosome. Because color-vision types can readily be estimated from allelic types of the M/LWS opsin gene, this polymorphic system offers researchers an excellent opportunity to study the association between vision and behavior. As a prerequisite for such studies, genetically determined color-vision types must be concordant with phenotypes determined directly by behavioral criteria (e.g., by a color discrimination test). However, such correlations between genotypes and phenotypes have been studied only for callitrichine species. Using genetic, electrophysiological, and behavioral approaches, we evaluated the color vision of brown capuchin monkeys (Cebus apella), a representative non-callitrichine model animal for physiology and behavior. Two allelic M/LWS opsins-P545 and P530-were identified in the studied captive population. Females had one or both of the alleles, and males had either one. The retinal sensitivity in P530 dichromats was short-wave shifted relative to that in P545 dichromats, whereas that in P530/P545 trichromats was between the two groups. In a discrimination task using Ishihara pseudo-isochromatic plates, P530/P545 trichromats were successful in discriminating stimuli that P530 and P545 dichromats were unable to discriminate. In a food-search task, P530/P545 trichromats were able to locate red targets among green distracters as quickly as among white distracters, whereas both types of dichromats took longer. These results demonstrate the mutual consistency between genotypes and phenotypes of color vision, and provide a solid genetic basis on which the ecology and evolution of color vision can be investigated. PMID- 16342071 TI - Recent advances in color vision research. AB - The remarkable variation in color vision both among and within primate species is receiving increasing attention from geneticists, psychophysicists, physiologists, and behavioral ecologists. It is known that color vision ability affects foraging behavior. Color vision is also likely to have implications for predation avoidance, social behavior, mate choice, and group dynamics, and should also influence the choice of stimuli for cognitive experiments. Therefore, understanding the color vision of a study species is important and of particular significance to scientists studying species with polymorphic color vision (most platyrrhines and some strepsirrhines). The papers in this issue were inspired by a symposium held during the 20th Congress of the International Primatological Society at Turin, Italy, in August 2004. The aim of the symposium was to bring together research from a range of disciplines, using recent methodological advances in molecular, modeling, and experimental techniques, to help elucidate the evolution, ecological importance, and distribution of color vision genotypes and phenotypes. The symposium achieved its aim, and as with most research in expanding disciplines, there are surprises and many questions still to be answered. Further advances will be made using a combination of different approaches involving analyses at the level of molecu1es, types of cell and neural networks, detailed and long-term field work, modeling, and carefully controlled experimentation. PMID- 16342072 TI - Sugar concentration of fruits and their detection via color in the Central American spider monkey (Ateles geoffroyi). AB - Although most arguments explaining the predominance of polymorphic color vision in platyrrhine monkeys are linked to the advantage of trichromacy over dichromacy for foraging for ripe fruits, little information exists on the relationship between nutritional reward and performance in fruit detection with different types of color vision. The principal reward of most fruits is sugar, and thus it seems logical to investigate whether fruit coloration provides a long-distance sensory cue to primates that correlates with sugar content. Here we test the hypothesis that fruit detection performance via trichromatic color vision phenotypes provides better information regarding sugar concentration than dichromatic phenotypes (i.e., is a color vision phenotype with sufficient red green (RG) differentiation necessary to "reveal" the concentration of major sugars in fruits?). Accordingly, we studied the fruit foraging behavior of Ateles geoffroyi by measuring both the reflectance spectra and the concentrations of major sugars in the consumed fruits. We modeled detection performance with different color phenotypes. Our results provide some support for the hypothesis. The yellow-blue (YB) color signal, which is the only one available to dichromats, was not significantly related to sugar concentration. The RG color vision signal, which is present only in trichromats, was significantly correlated with sugar content, but only when the latter was defined by glucose. There was in fact a consistent negative relationship between fruit detection performance and sucrose concentration, although this was not significant for the 430 nm and 550 nm phenotypes. The regular trichromatic phenotypes (430 nm, 533 nm, and 565 nm) showed higher correlations between fruit performance and glucose concentration than the other two trichromatic phenotypes. Our study documents a trichromatic foraging advantage in terms of fruit quality, and suggests that trichromatic color vision is advantageous over dichromatic color vision for detecting sugar rich fruits. PMID- 16342073 TI - Influence of stimuli size on color discrimination in capuchin monkeys. AB - Large-field trichromacy is a general feature of protanope and deuteranope humans, provided that the stimuli size extends to an 8 degrees visual angle. In this study we compared the performance of five male and three female tufted capuchin monkeys (Cebus apella) in discriminating pairs of Munsell color papers. Human subjects were also studied in two-choice discrimination tests, using the same stimuli and apparatus employed for the monkeys. The results show that although the dichromatic humans showed improved discrimination with larger versions of the stimuli, the dichromatic monkeys exhibited the same performance for both stimuli sizes. Thus, Cebus apella apparently do not present large-field trichromacy-at least for the conditions in the present experiments. PMID- 16342074 TI - Color vision pigment frequencies in wild tamarins (Saguinus spp.). AB - The adaptive importance of polymorphic color vision found in many New World and some prosimian primates has been discussed for many years. Polymorphism is probably maintained in part through a heterozygote advantage for trichromatic females, as such individuals are observed to have greater foraging success when selecting ripe fruits against a background of forest leaves. However, recent work also suggests there are some situations in which dichromatic individuals may have an advantage, and that variation in color vision among individuals possessing different alleles may also be significant. Alleles that confer a selective advantage to individuals are expected to occur at a higher frequency in populations than those that do not. Therefore, analyzing the frequencies of color vision alleles in wild populations can add to our understanding of the selective advantages of some color vision phenotypes over others. With this aim, we used molecular techniques to determine the frequencies of color vision alleles in 12 wild tamarin groups representing three species of the genus Saguinus. Our results show that allele frequencies are not equal, possibly reflecting different selective regimes operating on different color vision phenotypes. PMID- 16342075 TI - Color vision in marmosets and tamarins: behavioral evidence. AB - Here we demonstrate differences in the relative performance of 15 callitrichids tested in a series of color visual discrimination experiments. Munsell color chips were chosen as stimuli based on their use in earlier experiments with human dichromats. We show behavioral evidence for the existence of four distinct kinds of color-vision phenotypes, each of which has slightly different color discrimination abilities. The different phenotypes may offer different advantages. The data are in accordance with the existence of a visual polymorphism in callitrichids. PMID- 16342076 TI - Comparative use of color vision for frugivory by sympatric species of platyrrhines. AB - Ateles spp. and Alouatta spp. are often sympatric, and although they are mainly frugivorous and folivorous, respectively, they consume some of the same fruit species. However, they differ in terms of color vision, which is thought to be important for fruit detection. Alouatta spp. have routine trichromatic color vision, while Ateles spp. presents the classic polymorphism of platyrrhines: heterozygous females have trichromatic color vision, and males and homozygous females have dichromatic vision. Given these perceptual differences, one might expect Alouatta spp. to consume more reddish fruits than Ateles spp., since trichromats have an advantage for detecting fruits of that hue. Furthermore, since Ateles spp. have up to six different color vision phenotypes, as do most other platyrrhines, they might be expected to include fruits with a wider variety of hues in their diet than Alouatta spp. To test these hypotheses we studied the fruit foraging behavior of sympatric Alouatta palliata and Ateles geoffroyi in Costa Rica, and modeled the detectability of fruit via the various color vision phenotypes in these primates. We found little similarity in fruit diet between these two species (Morisita = 0.086). Furthermore, despite its polymorphism, A. geoffroyi consumed more reddish fruits than A. palliata, which consumed more greenish fruits. Our modeling results suggest that most fruit species included in the diet of A. geoffroyi can be discriminated by most color vision phenotypes present in the population. These findings show that the effect of polymorphism in platyrrhines on fruit detection may not be a disadvantage for frugivory. We suggest that routine trichromacy may be advantageous for other foraging tasks, such as feeding on young leaves. PMID- 16342077 TI - Cranial neural crest cell migration in cockatiel Nymphicus hollandicus (Aves: Psittaciformes). AB - Parrots have developed unique jaw muscles in their evolutionary history. The M. pseudomasseter, which completely covers the lateral side of the jugal bar, is regarded as a jaw muscle unique to parrots. In a previous study, I presented a hypothesis on the relevance of modifications in the regulation of cranial neural crest cell (NCC) development to the generation of this novel jaw muscle based on histological analyses (Tokita [2004] J Morphol 259:69-81). In the present study, I investigated distribution and migration patterns of cranial neural crest cells (NCCs) through parrot embryogenesis with immunohistochemical techniques to further understand the role of cranial NCCs in the evolution of the M. pseudomasseter, and to provide new information on the relative plasticity in cranial NCC migration at early stages of avian development. The basic nature of cranial NCC development was mostly conserved between chick and parrot. In both, cranial NCCs migrated from the dorsal tip of the neural tube in a ventral direction. Three major populations were identified in their cranial NCCs. Migration pathways of these cells were almost identical between chick and parrot. The principal difference was seen in the relative timing of cranial NCC migration. In the parrot, cranial NCC migration into the first pharyngeal arch was more advanced than in the chick at early stages of development. Such a temporal shift in cranial NCC migration might influence architectural patterning of parrot jaw muscles that generates new muscle like M. pseudomasseter. PMID- 16342078 TI - Scaling of body frontal area and body width in birds. AB - By analyzing a homogenous dataset we show, in contradiction to a previous study, that the scaling of body frontal area (S(b)) with body mass (m(b)) does not differ between passerine and nonpasserine birds. It is likely that comparison of data collected from live passerines with data collected from frozen nonpasserines had led to the incorrect conclusion that the scaling of S(b) varied between the taxa. We suggest that body dimensions collected from frozen specimens, or specimens stored in alcohol, are not applicable to live birds, and that both the current equations presented in the literature for predicting S(b) from m(b) may lead to inaccurate estimates. Using data from preserved specimens, we found that S(b) scales isometrically with m(b) (S(b) proportional, variant m(b) (0.66)), and therefore we found no evidence for larger birds being more streamlined than smaller birds. S(b) scales with negative allometry against wingspan (b), however, and b scales with positive allometry against m(b), so larger birds have smaller S(b) relative to b. In addition, it appears that dorsoventral flattening of the body is a general characteristic of bird's bodies but that it is more pronounced in larger birds, suggesting perhaps a function in terms of increased lift during forward flight. It appears that bird's bodies obey the surface-to-area geometric scaling law, but bird body shape may vary in relation to aerodynamic function. We suggest that a large-scale study, simultaneously measuring S(b) and m(b) in live passerines and nonpasserines, is required to improve the predictive power of S(b) upon m(b) scaling equations, which play a key role in the estimation of mechanical power consumption in flight in birds. Furthermore, the relations between bird body shape and axial skeleton dimensions, with reference to aerodynamic adaptation, warrant further investigation. PMID- 16342079 TI - Guidelines for improving the reproducibility of quantitative multiparameter immunofluorescence measurements by laser scanning cytometry on fixed cell suspensions from human solid tumors. AB - BACKGROUND: Laser scanning Cytometry (LSC) is a versatile technology that makes it possible to perform multiple measurements on individual cells and correlate them cell by cell with other cellular features. It would be highly desirable to be able to perform reproducible, quantitative, correlated cell-based immunofluorescence studies on individual cells from human solid tumors. However, such studies can be challenging because of the presence of large numbers of cell aggregates and other confounding factors. Techniques have been developed to deal with cell aggregates in data sets collected by LSC. Experience has also been gained in addressing other key technical and methodological issues that can affect the reproducibility of such cell-based immunofluorescence measurements. METHODS AND RESULTS: We describe practical aspects of cell sample collection, cell fixation and staining, protocols for performing multiparameter immunofluorescence measurements by LSC, use of controls and reference samples, and approaches to data analysis that we have found useful in improving the accuracy and reproducibility of LSC data obtained in human tumor samples. We provide examples of the potential advantages of LSC in examining quantitative aspects of cell-based analysis. Improvements in the quality of cell-based multiparameter immunofluorescence measurements make it possible to extract useful information from relatively small numbers of cells. This, in turn, permits the performance of multiple multicolor panels on each tumor sample. With links among the different panels that are provided by overlapping measurements, it is possible to develop increasingly more extensive profiles of intracellular expression of multiple proteins in clinical samples of human solid tumors. Examples of such linked panels of measurements are provided. CONCLUSIONS: Advances in methodology can improve cell-based multiparameter immunofluorescence measurements on cell suspensions from human solid tumors by LSC for use in prognostic and predictive clinical applications. PMID- 16342080 TI - Electronic structures of 3d-metal mononitrides. AB - Bond distances, vibrational frequencies, electron affinities, ionization potentials, and dissociation energies of the title molecules in neutral, positively, and negatively charged ions were studied by use of density functional methods B3LYP, BLYP, BHLYP, BPW91, and B3PW91. The calculated results are compared with experiments and previous theoretical studies. It was found that the calculated properties are highly dependent on the functionals employed, in particular for the dissociation energy and vibrational frequency. For neutral species, pure density functional methods BLYP and BPW91 have relatively good performance in reproducing the experimental bond distance and vibrational frequency. For cations, hybrid exchange functional methods B3LYP and B3PW91 are good in predicting the dissociation energy. For both neutral and charged species, BHLYP tends to give smaller dissociation energy. PMID- 16342081 TI - Activation barriers for DNA alkylation by carcinogenic methane diazonium ions. AB - Methylation reactions of the DNA bases with the methane diazonium ion, which is the reactive intermediate formed from several carcinogenic methylating agents, were examined. The SN2 transition states of the methylation reactions at N7, N3, and O6 of guanine; N7, N3, and N1 of adenine; N3 and O2 of cytosine; and O2 and O4 of thymine were calculated using the B3LYP density functional method. Solvation effects were examined using the conductor-like polarizable continuum method and the combined discrete/SCRF method. The transition states for reactions at guanine N3, adenine N7, and adenine N1 are influenced by steric interactions between the methane diazonium ion and exocyclic amino groups. Both in the gas phase and in aqueous solution, the methylation reactions at N atoms have transition states that are looser, and generally occur earlier along the reaction pathways than reactions at O atoms. The forming bonds in the transition states in water are 0.03 to 0.13 A shorter than those observed in the gas phase, and the activation energies are 13 to 35 kcal/mol higher. The combined discrete/SCRF solvation energy calculations using base-water complexes with three water molecules yield base solvation energies that are larger than those obtained from the CPCM continuum method, especially for cytosine. Reactivities calculated using barriers obtained with the discrete/SCRF method are consistent with the experimentally observed high reactivity at N7 of guanine. PMID- 16342083 TI - Preliminary results of integrated therapy for patients with knee osteoarthritis. AB - OBJECTIVE: To investigate the effects of integrated therapy on the functional status of patients with knee osteoarthritis (OA). METHODS: A total of 140 subjects with bilateral knee OA (Altman grade II) were randomized sequentially into 4 groups (groups I-IV). Group I received isokinetic exercises; group II received isokinetic exercise and pulse ultrasound for periarticular soft tissue pain; group III received isokinetic exercise, pulse ultrasound, and intraarticular hyaluronan therapy; and group IV acted as the control group. The therapeutic effects of the interventions were evaluated by changes in Lequesne's index, knee range of motion, peak muscle torques of knee flexion and extension, and ambulation speed after 8 weeks of treatment and at followup 1 year later. In addition, changes in visual analog scale pain and rates of attrition in each group were also recorded. RESULTS: Patients in groups I-III exhibited increased muscle peak torques and significantly reduced pain and disability after treatment and at followup. Groups II and III showed significant improvements in range of motion and ambulation speed after treatment. Group III also showed the greatest increase in walking speed and decrease in disability after treatment and at followup. Both group II and group III had significant gains in muscular strength after treatment and at followup; group III showed the greatest gains. CONCLUSION: An integrated therapy deals with the extra- and intraarticular progressive pathologic changes, and kinesiologic management of OA is suggested for the management of knee OA. PMID- 16342084 TI - A disease-specific self-help program compared with a generalized chronic disease self-help program for arthritis patients. AB - OBJECTIVE: Both the Arthritis Self-Management Program (ASMP) and the generic Chronic Disease Self-Management Program (CDSMP) have been shown to be successful in improving conditions in patients with arthritis. This study compared the relative effectiveness of the 2 programs for individuals with arthritis. METHODS: Patients whose primary disease was arthritis were randomized to the ASMP (n = 239) or to the CDSMP (n = 116). Analyses of covariance were used to compare the outcome measures for the 2 programs at 4 months and 1 year. Measures included quality of life outcomes (self reported, health distress, disability, activity limitation, global health, pain, and fatigue), health behaviors (practice of mental stress management, stretching and strength exercise, aerobic exercise), self efficacy, and health care utilization (physician visits and hospitalizations). RESULTS: Both programs showed positive results. The disease specific ASMP appeared to have advantages over the more generic CDSMP for patients with arthritis at 4 months. These advantages had lessened slightly by 1 year. CONCLUSION: The disease-specific ASMP should be considered first where there are sufficient resources and participants. However, both programs had positive effects, and the CDSMP should be considered a viable alternative. PMID- 16342085 TI - Longitudinal study of negative workplace events among employed rheumatoid arthritis patients and healthy controls. AB - OBJECTIVE: Most studies of employment in patients with rheumatoid arthritis (RA) have focused on job loss. Less is known about workplace events in patients who continued to work. The goal of this longitudinal study was to compare the incidence of negative workplace events between employed patients with RA and healthy controls. METHODS: Participants completed the work domains of the Psychiatric Epidemiology Research Interview Life Events Scale and the Inventory of Small Life Events Scale measuring major and minor workplace events. Events were compared between groups according to psychosocial, clinical, and job characteristics. RESULTS: A total of 122 patients with RA and 122 healthy controls were enrolled with similar demographic and occupational characteristics. There were no differences in percentages of patients and controls who had at least 1 major (35% versus 31%) or 1 minor (48% versus 55%) negative event. For patients with RA, negative events were associated with having more pain, more fatigue, more social stress, and less job autonomy (P < or = 0.05). For controls, in addition to social stress, negative events were associated with job characteristics, functional status, and social support (P < or = 0.05). CONCLUSION: A comparable number of patients with RA and controls had negative workplace events. In addition to the well-known contributions of job autonomy and pain, social stress and fatigue also were found to be important variables related to negative events in patients with RA. These potentially modifiable variables have not been fully evaluated with respect to long-term employment in these patients. PMID- 16342086 TI - Frequency of microangiopathic hemolytic anemia in patients with systemic lupus erythematosus exacerbation: Distinction from thrombotic thrombocytopenic purpura, prognosis, and outcome. PMID- 16342088 TI - Translating research into practice: can we get there from here? PMID- 16342089 TI - Comparison of the COX-inhibiting nitric oxide donator AZD3582 and rofecoxib in treating the signs and symptoms of Osteoarthritis of the knee. AB - OBJECTIVE: To compare the efficacy, safety, and tolerability of AZD3582 with that of rofecoxib, naproxen, and placebo in patients with osteoarthritis (OA) of the knee, and to define the dosage of AZD3582 (125 mg, 375 mg, and 750 mg twice a day) that is noninferior in efficacy to rofecoxib. METHODS: A double-blind study of 672 patients with OA of the knee was conducted. Patients who experienced increased pain on withdrawal of analgesia were randomized to receive AZD3582 125 mg, 375 mg, or 750 mg twice a day; rofecoxib 25 mg once a day; naproxen 500 mg twice a day; or placebo for 6 weeks. Efficacy, tolerability, and safety were monitored throughout the study. The primary variable was the change in Western Ontario and McMaster Universities Osteoarthritis Index pain subscale from baseline to the mean of weeks 4 and 6, comparing AZD3582 with placebo for superiority and with rofecoxib for noninferiority using a predefined margin of 10 mm. RESULTS: For the primary variable, AZD3582 375 mg and 750 mg were superior to placebo (least squares mean difference [95% confidence interval] -12 mm [-18, 6], P < 0.001 and -13 mm [-19, -7], P < 0.001, respectively) and were noninferior to rofecoxib (-2 mm [-8, 4], P < 0.001 and -3 mm [-9, 3], P < 0.001, respectively). AZD3582 125 mg was not significantly different from placebo for the primary variable. CONCLUSION: AZD3582 375 mg and 750 mg twice a day were superior to placebo and as effective as rofecoxib 25 mg/day in treating the signs and symptoms of OA of the knee. AZD3582 125 mg twice a day was not statistically different from placebo. PMID- 16342090 TI - Diagnostic value of single-photon-emission computed tomography in severe central nervous system involvement of systemic lupus erythematosus: a case-control study. AB - OBJECTIVE: To evaluate the diagnostic value of single-photon-emission computed tomography (SPECT) in severe central nervous system (CNS) involvement of systemic lupus erythematosus (SLE). METHODS: Forty-three patients with SLE, including 22 with CNS-SLE and 21 with non-CNS-SLE, underwent SPECT and magnetic resonance imaging (MRI) examinations. SPECT was repeated 1-2 months after treatment in patients with abnormal findings. RESULTS: SPECT and MRI abnormalities were detected in 20 (90.9%) and 10 (45.5%) of the 22 patients with CNS-SLE, respectively (P < 0.01). For 4 patients with cerebral infarction or hemorrhage, SPECT was equally as sensitive as MRI (100%). For the patients with CNS-SLE with diffuse presentations, SPECT was more sensitive than MRI in revealing abnormalities (16 [88.9%] of 18 patients versus 6 [33.3%] of 18 patients; P < 0.01). In 19 (95.0%) patients, the abnormal SPECT finding manifested as moderate to severe perfusion defect (15 in frontal lobe, 11 in parietal lobe, 11 in basal ganglia, 3 in temporal lobe, and 17 in multiple regions). Although mild perfusion defect was also detected in 4 (19.0%) of the patients with non-CNS-SLE, it only involved a single region and spared the frontal and parietal lobes. Repeated SPECT after treatment showed that perfusion defect had improved significantly or even disappeared in 11 (84.6%) of 13 patients with diffuse CNS-SLE with abnormal findings before treatment. CONCLUSION: Moderate to severe perfusion defect in SPECT involving multiple regions, especially in the frontal and parietal lobes and basal ganglia, in patients with lupus suggests CNS involvement. SPECT is more sensitive than MRI in revealing damage in diffuse CNS-SLE, and is useful in followup, especially for monitoring disease severity and guiding treatment. PMID- 16342091 TI - Incidence and prevalence of ankylosing spondylitis in Northern Norway. AB - OBJECTIVE: To determine the incidence and prevalence of ankylosing spondylitis (AS) over a prolonged period in the 2 northernmost counties of Norway, where HLA B27 has a high prevalence in the population. METHODS: We conducted a cohort study of all patients registered with a diagnosis of AS between 1960 and 1993 at the University Hospital of Northern Norway, which is the sole rheumatology department serving these counties. We registered demographics, year of disease onset (clinical disease), and year of diagnosis (radiograph confirmation) for all patients. The date of onset of clinical disease in patients with AS was used in the calculation of incidence rates. Annual incidence and point/period prevalence rates were expressed per 100,000 adults. Primary AS was defined as AS in the absence of psoriasis or inflammatory bowel disease (IBD). RESULTS: A total of 534 patients (75.1% male, mean age at clinical diagnosis 24.2 years, 93.0% HLA-B27 positive) had a confirmed diagnosis of AS (by the modified New York criteria). Median time from disease onset to radiologic confirmation was 8.0 years. Annual incidence of primary AS (n = 417) was 7.26, while estimated point prevalence rose from 0.036% in 1970 to 0.10% in 1980 and to 0.21% in 1990 with a period prevalence of 0.26%. AS was secondary to psoriasis or IBD in 117 patients (18.1%), with a diagnostic delay similar to that in primary AS. Annual incidence (14.1) and period prevalence in 1982-1993 (0.41%) were significantly higher in the town of Tromso than in the surrounding rural region (5.21 and 0.22%, respectively). Mortality in patients with AS was low. CONCLUSION: The incidence of AS was relatively stable in the northern part of Norway over a 34-year period. Incidence and prevalence are higher than reported in similar studies from Finland and Minnesota, possibly due to a higher population prevalence of HLA-B27. PMID- 16342092 TI - Antinuclear antibody titer and antibodies to extractable nuclear antigens. PMID- 16342093 TI - Outcome of patients with active ankylosing spondylitis after two years of therapy with etanercept: clinical and magnetic resonance imaging data. AB - OBJECTIVE: To examine the long-term outcome of patients with active ankylosing spondylitis (AS) clinically and by magnetic resonance imaging (MRI) after continuous treatment with the tumor necrosis factor (TNF) receptor fusion protein etanercept over 2 years. METHODS: Overall, 26 patients with active AS were treated with etanercept 25 mg twice daily subcutaneously, twice weekly with no concomitant disease-modifying antirheumatic drugs (DMARDs) or steroids. The clinical response was assessed by standardized parameters. Inflammatory spinal lesions were quantified by the ASspiMRI-a rating gadolinium-enhanced (T1-weighted gadolinium diethylenetriaminepentaacetic acid) and STIR MRI sequences. The primary outcome was a Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) improvement > or =50% after 2 years of etanercept therapy compared with the baseline value of the study. RESULTS: Overall, 21 (70%) of 30 patients completed year 2. In the intent-to-treat analysis, 54% of the patients showed a 50% improvement according to the BASDAI and a 40% improvement according to the Assessment in Ankylosing Spondylitis (ASAS) criteria. In the completer analysis, 9 (43%) of 21 patients were in partial remission according to ASAS criteria. Mean +/- SD BASDAI scores, which were elevated at baseline (6.3 +/- 1.6), remained low: 2.7 +/- 2.4 after 2 years compared with 2.6 +/- 2.2 at week 54. In accordance, all other clinical parameters showed sustained improvement during year 2. The majority of patients had no disease activity flares. MRI evaluation showed a 75% improvement of active spinal lesions, but minor spinal inflammation was still present in 64% of the patients after 2 years. There were 2 serious adverse events leading to discontinuation of etanercept. CONCLUSION: The clinical efficacy and safety of etanercept in patients with active AS without simultaneous administration of DMARDs or steroids over 2 years of continuous treatment is confirmed. Spinal inflammation as depicted by MRI decreased significantly, but a few patients still had some spinal inflammation even after long-term anti-TNF therapy. PMID- 16342094 TI - Measures matter: racial disparities in the provision of total knee replacement. PMID- 16342095 TI - Pattern of infliximab utilization in rheumatoid arthritis patients at an academic medical center. AB - OBJECTIVE: To investigate the pattern of use of infliximab with an emphasis on treatment escalation and the durability of infliximab use in the management of rheumatoid arthritis (RA) in an academic setting. METHODS: We conducted a retrospective review of pharmacy and medical records of 183 patients with RA who received at least 1 infliximab infusion at the infusion centers of the Brigham and Women's Hospital. Treatment escalation was defined as an increase in the dosage of infliximab to >3 mg/kg and/or a decrease in the dosing interval to <7 weeks between infusions. RESULTS: A total of 183 patients with RA received infliximab infusions for a mean +/- SD duration of 58.2 +/- 56.6 weeks. Infliximab was discontinued in 48% of the patients during the first year of therapy and in 67% of the patients overall. A total of 126 patients had a treatment escalation, including 25 patients with a dose increase, 35 patients with a decrease in the interval, and 66 patients with both. Infliximab treatment was associated with a decrease in corticosteroid and methotrexate doses. Patients who had a treatment escalation were more likely to continue infliximab infusions compared with patients without a treatment escalation (odds ratio 2.0, 95% confidence interval 1.0-4.1). CONCLUSION: The use of infliximab may be an effective treatment for RA; however, a substantial number of patients will discontinue its use. Treatment escalation is commonly used in the management of RA with infliximab and is associated with longer duration of infliximab use. PMID- 16342096 TI - Effect of physical activity on functional status among older middle-age adults with arthritis. AB - OBJECTIVE: To determine the effect of leisure time and work-related physical activity on changes in physical functioning among 3,554 nationally representative survey respondents, ages 53-63 years in 1994, with arthritis and joint symptoms, interviewed in the Health and Retirement Study (HRS). METHODS: In 1992-1994, light and vigorous exercise items were empirically categorized into recommended, insufficient, and inactive leisure time physical activity levels using data from the HRS. Leisure and work-related physical activity levels in 1994 were used to predict 1996 functional decline or improvement, controlling for baseline functional difficulties, health status, sociodemographic characteristics, and behavioral risk factors. RESULTS: Whereas 29.7% of respondents reported functional declines in 1996, 38.6% of those with baseline difficulties in 1994 reported improvement. Compared with inactive respondents, recommended and insufficient leisure time physical activity were equally protective against functional decline (odds ratio [OR] 0.59 and 0.62, respectively; P < 0.0001). Higher levels of physical activity were also modestly associated with functional improvement among respondents with baseline functional difficulties (OR 1.47, P = 0.05 and OR 1.45, P = 0.01, respectively). Work-related physical activity was not a significant predictor of decline or improvement. CONCLUSION: Given the high prevalence of arthritis, even modest increases in rates of lifestyle physical activity among older adults could make a substantial contribution to disability free life expectancy. PMID- 16342097 TI - Activity limitation in rheumatoid arthritis correlates with reduced grip force regardless of sex: the Swedish TIRA project. AB - OBJECTIVE: To evaluate activity limitations 3 years after diagnosis of early rheumatoid arthritis (RA) in relation to grip force and sex. METHODS: A total of 217 patients, 153 women and 64 men, with recent-onset RA were included. Activity limitations were reported using the Health Assessment Questionnaire (HAQ) and the Evaluation of Daily Activities Questionnaire (EDAQ). The relationships between activity limitations versus grip force (measured by the Grippit), walking speed, functional impairment, grip ability, pain, plasma C-reactive protein, the 28 joint disease activity score and its components, the physician's global assessment of disease activity, and sex were analyzed by partial least squares (PLS). RESULTS: Women had significantly lower grip force and more activity limitations (HAQ and EDAQ) than men. The PLS analyses demonstrated that grip force was the strongest regressor of activity limitation, closely followed by walking speed. However, within subgroups based on grip force (group 1 = grip force <114 N, group 2 = 116-206 N, group 3 = 214-321 N, group 4 = grip force >328 N) and including sexes, women and men had corresponding degrees of activity limitation as reported by the HAQ and EDAQ. CONCLUSION: Our results indicate that the more pronounced activity limitations seen in women with RA, as compared with men, may be explained by lower grip force rather than sex. PMID- 16342098 TI - Pain as a mediator of sleep problems in arthritis and other chronic conditions. AB - OBJECTIVE: To examine the associations between arthritis and insomnia symptoms and unrefreshing sleep, as well as the role of pain as a mediator of these relationships. METHODS: Analyses were conducted on the cross-sectional, nationally representative, weighted sample of adults > or =18 years of age (n = 118,336) in the 2000/2001 Canadian Community Health Survey. Four logistic regression models were estimated for each sleep problem (model 1: arthritis only; model 2: model 1 + sociodemographic characteristics, lifestyle factors, and other chronic conditions; model 3: model 2 + mental health [stress, depression]; and model 4: model 3 + pain). Mediation by pain was quantified by the percentage change in the effect of arthritis on a particular sleep problem by comparing models 3 and 4. RESULTS: The prevalence of insomnia symptoms and unrefreshing sleep in persons with arthritis was 24.8% and 11.9%, respectively. These estimates are twice as high as those for persons without arthritis. In multivariate regression analyses, the addition of pain decreased the effect of arthritis by 53% (insomnia symptoms) and 64% (unrefreshing sleep). The effect of arthritis was still statistically significant in these models, suggesting that pain is a partial mediator of these relationships. CONCLUSION: Insomnia symptoms and unrefreshing sleep affect a considerable proportion of individuals with arthritis. Pain mediates a substantial amount of the relationship between arthritis and sleep problems. Better pain management could significantly improve sleep in individuals with arthritis. PMID- 16342099 TI - Use of self-management behaviors to cope with rheumatoid arthritis stressors. AB - OBJECTIVE: To examine self-management behaviors used to cope with 5 rheumatoid arthritis (RA)-related stressors (pain, fatigue, physical limitations, joint changes, and symptom unpredictability) and the relationship between use of self management behaviors and ability to perform life activities. METHODS: Data were from telephone interviews of 511 persons with RA. Participants were presented with lists of self-management behaviors for each stressor and asked to indicate which they had used in the past year. Performance of life activities was assessed concurrently (baseline) and 1 year later (followup). The number of activities affected (difficulty performing or unable to perform) and the number that individuals were completely unable to perform were calculated. Multiple regression analyses were used to examine the association of perceived stressor impact and arthritis education with strategy use and the association of self management behaviors with performance of life activities. RESULTS: Four self management strategies were identified: accommodation, active remediation, social, and perseverance. Greater perceived impact of stressors was associated with use of more self-management strategies in total for each stressor (beta = 0.43-0.73, all P < 0.0001) and with use of accommodation (beta = 0.26-0.29, P < 0.0001), social (beta = 0.23-0.43, P < 0.0001), and perseverance (beta = 0.29-0.46, P < 0.0001) strategies. Greater use of accommodation strategies at baseline was associated with an increase in the number of activities affected at followup (beta = 0.11, P < 0.05); greater use of perseverance strategies was associated with a decrease in activities unable to perform (beta = -0.13, P < 0.01). CONCLUSION: Self-management behaviors are commonly used and may help prevent loss of the ability to perform life activities. PMID- 16342100 TI - Assessment of inflammatory response in a study of uveitis associated with juvenile idiopathic arthritis: comment on the article by Smith et al. PMID- 16342101 TI - Positive association between serum 25-hydroxyvitamin D level and bone density in osteoarthritis. AB - OBJECTIVE: To describe the association between serum 25-hydroxyvitamin D (25[OH]D) level and bone mineral density (BMD) in persons with primary knee osteoarthritis (OA). METHODS: We conducted a population-based survey of the Framingham Study. A total of 228 subjects with primary radiographic knee OA were identified. For vitamin D status, 25(OH)D levels < or =15 ng/ml were classified as vitamin D deficient, 25(OH)D levels 16-32 ng/ml were classified as hypovitaminosis D, and 25(OH)D levels >32 ng/ml were classified as vitamin D replete. We compared average BMD between categories of 25(OH)D levels in subjects with OA using a linear regression model while adjusting for sex, age, body mass index (BMI), knee pain, physical activity, cohort, and disease severity. RESULTS: Mean age was 74.4 years and 36% were men. Of 228 individuals, 15% were vitamin D deficient, 51% had hypovitaminosis D, and 34% were vitamin D replete. Compared with subjects with vitamin D deficiency, those with hypovitaminosis D had a 7.3% higher BMD (adjusted percent difference; P = 0.02) and vitamin D replete subjects had an 8.5% higher BMD (adjusted percent difference; P = 0.02; test for trend across categories: P = 0.04). CONCLUSION: We observed a significant positive association between serum 25(OH)D and BMD in individuals with primary knee OA, independent of sex, age, BMI, knee pain, physical activity, and disease severity. Given the high prevalence of low 25(OH)D status in persons with knee OA and the positive association between 25(OH)D and BMD, vitamin D supplementation may enhance BMD in individuals with OA. PMID- 16342102 TI - Effects of supervised cardiovascular training program on exercise tolerance, aerobic capacity, and quality of life in patients with systemic lupus erythematosus. AB - OBJECTIVE: To determine if supervised cardiovascular training improves exercise tolerance, aerobic capacity, depression, functional capacity, and quality of life in patients with systemic lupus erythematosus (SLE). METHODS: Sixty women with SLE (ages 18-55 years) were evaluated using Short Form 36, visual analog scale for pain, scale for fatigue, Beck Depression Inventory, and Health Assessment Questionnaire (HAQ), and participated in a training protocol of incremental load on a treadmill with computed gas metabolic analysis. Maximum oxygen consumption (VO(2max)) and anaerobic threshold VO(2) were calculated with a SensorMedics Vmax29C analyzer (Sensor Medics, Yorba Linda, CA), and heart rate was measured by electrocardiogram. Patients were divided into 2 groups: a training group (41 patients) that participated in the supervised cardiovascular training program and a control group (19 patients) that did not participate in the program. All variables were analyzed at baseline and after 12 weeks for both groups. The training program occurred in the morning for 60 minutes, 3 times a week for 12 weeks. Statistical analysis included Wilcoxon's rank sum test, Mann-Whitney U test, chi-square test, and Fisher's exact test. P values <0.05 were considered to be statistically significant. RESULTS: The 2 groups were homogeneous and comparable at baseline. The training group showed a significant improvement of aerobic capacity measured by anaerobic threshold VO(2) (14.67 +/- 3.03 versus 17.08 +/- 3.35 ml/kg/minute, P < 0.001). Comparison of the training group and control group after 12 weeks showed a significant difference relating to VO(2max) (24.31 +/- 4.61 versus 21.21 +/- 3.88 ml/kg/minute, P = 0.01) and anaerobic threshold VO(2) (17.08 +/- 3.35 versus 13.66 +/- 2.82 ml/kg/minute, P < 0.0001). After cardiovascular training, we found a significant improvement of Beck inventory score (8.37 +/- 12.79 versus 2.90 +/- 3.00, P < 0.001) and HAQ score (0.14 +/- 0.21 versus 0.06 +/- 0.19, P < 0.01) in the training group. CONCLUSION: This study showed significant improvement in exercise tolerance, aerobic capacity, quality of life, and depression after a supervised cardiovascular training program in patients with SLE. PMID- 16342103 TI - Clinical responsiveness of self-report functional assessment measures for children with juvenile idiopathic arthritis undergoing intraarticular corticosteroid injections. AB - OBJECTIVE: The Childhood Health Assessment Questionnaire (CHAQ), Juvenile Arthritis Functional Assessment Report (JAFAR), and Juvenile Arthritis Functional Status Index (JASI) are widely used functional measures for juvenile idiopathic arthritis (JIA) that differ in content, format, and completion time. We compared the responsiveness and child-parent agreement of the JAFAR, CHAQ, and JASI in a prospective, multicenter study. METHODS: Children and adolescents from 5 rheumatology centers were enrolled. Subjects were about to undergo therapy (intraarticular corticosteroid injections [IAS] and methotrexate or hip surgery (MTX/hip]) expected to produce a functional improvement. All subjects were studied before the intervention and at 6 weeks and 6 months posttreatment. At each study visit, the 3 measures were administered in randomized, balanced order to both parents and children. RESULTS: A total of 92 subjects (mean age 12.8 years) were enrolled in the study, 74 of which were in the IAS group. The responsiveness of all 3 measures was moderate to strong. The standardized response mean at 6 weeks for the IAS group on the JAFAR, CHAQ, and JASI was 0.41 (95% confidence interval [95% CI] 0.18, 0.64), 0.70 (95% CI 0.47, 0.93), and 0.36 (95% CI 0.13, 0.59), respectively. The CHAQ was somewhat more responsive to change at 6 weeks (IAS group: relative efficiency 0.34 [JAFAR], 0.27 [JASI]), but less responsive at 6 months (MTX/hip group: relative efficiency 5.1 [JAFAR], 3.9 [JASI]). All 3 questionnaires showed acceptable parent-child agreement, and overall, there were few differences between the 3 questionnaires. CONCLUSION: The functional outcome measures currently used for JIA are all adequately responsive for use in trials or in the clinic setting. The choice of which measure to use should therefore be based on the time available for completion, the intended clinical/research use, and the depth of content required. PMID- 16342104 TI - Electronically monitored adherence to medications by newly diagnosed patients with juvenile rheumatoid arthritis. AB - OBJECTIVE: To describe patterns of adherence to nonsteroidal antiinflammatory drugs (NSAIDs) in newly diagnosed patients with juvenile rheumatoid arthritis (JRA), and to examine demographic and disease-related variables as potential predictors of adherence. METHODS: Adherence to NSAIDs was monitored in 48 children with JRA (mean age 8.6 years) over 28 consecutive days using an electronic monitoring device. Measures of disease activity (active joint counts, morning stiffness) and demographics (age, sex, ethnicity, socioeconomic status) were also obtained. RESULTS: Using an 80% adherence cut point, 25 (52%) patients were classified as adherent and 23 (48%) as nonadherent. There was considerable variability across patients, with full adherence (taking all doses on time) ranging from 0 to 100% of the monitored days. Logistic regression showed that active joint count and socioeconomic status were the only significant predictors. Both were positively related to adherence. The model correctly classified 70.5% of patients as either adherent or nonadherent (Cox and Snell R(2) = 0.295, P = 0.0005). CONCLUSION: Children newly diagnosed with JRA are more likely to adhere to an NSAID regimen if they have a greater number of active joints or their families have higher socioeconomic status. The former finding suggests that children's adherence is symptom-driven, while the latter suggests that families of lower socioeconomic status deserve special attention to address adherence issues. PMID- 16342105 TI - Impact of shoulder, elbow, and knee joint involvement on assessment of rheumatoid arthritis using the American College of Rheumatology Core Data Set. AB - OBJECTIVE: To determine the most sensitive scoring method for assessment of rheumatoid arthritis (RA) disease activity using the American College of Rheumatology Core Data Set. METHODS: The subjects were 4,530 patients with RA (mean age 57.9 years, mean disease duration 12.7 years) who participated in a large observational cohort study of RA patients. The 68 joints assessed were classified into 15 joint areas, and each joint variable was categorized based on the presence or absence of swelling or pain in these areas. Multiple linear regression and analysis of variance were used to evaluate the significance of effects of these 15 joint areas on variables for assessment of RA disease activity such as patient's assessment of pain on a visual analog scale (VAS), patient's and physician's global assessment of disease activity on a VAS, HAQ (Health Assessment Questionnaire), and Japanese HAQ. RESULTS: Although the 3 most frequently affected joints were the wrist, metacarpophalangeal joints, and proximal interphalangeal joints, the 5 joints with the largest contributions to all of the variables assessed for disease activity were the shoulder, elbow, and knee joints, followed by the wrist and ankle joints. The combination of shoulder, elbow, and knee joints accounted for approximately 70% of the contribution to all the variables, while addition of the wrist and ankle joints increased this value to approximately 90%. CONCLUSION: Scoring for assessment of RA disease activity would be more sensitive if separate joints such as the shoulder, elbow, knee, wrist, and ankle joints were weighted differently. PMID- 16342106 TI - Course and prognosis of knee complaints in general practice. AB - OBJECTIVE: Patients frequently present with knee complaints in general practice. Information about the course and prognosis of knee complaints is needed to inform patients and facilitate decisions on referral and treatment. The objective of the study was to assess the course of knee complaints and to identify predictors of outcome in patients visiting their general practitioner with a new episode of knee complaints. METHODS: Data were collected by means of self-administered questionnaires. After 3 and 12 months of followup, the following outcomes were assessed: perceived recovery, change in pain, and change in physical functioning. As potential predictors of outcome, several sociodemographic variables, characteristics of the symptom, baseline scores of the outcome measures, and intra- and extra-individual variables were analyzed using multiple regression analyses. RESULTS: We included 251 patients with a new episode of knee complaints presented in general practice. Only 25% reported recovery after 3 months, increasing to 44% after 12 months. A history of knee complaints, a longer duration of the current episode of knee complaints, other coexisting musculoskeletal complaints, and a higher level of distress were associated with a worse prognosis. In the linear regression models, 41-53% of the variance in pain reduction and improvement in functioning could be explained by the predictors. The area under the receiver operating characteristic curves, estimating the predictive accuracy of the Cox regression models concerning perceived recovery, was 0.77 after 3 months and 0.72 after 12 months. CONCLUSION: Many patients did not recover after 12 months. Distress was found to be strongly associated with less pain reduction and less improvement in functioning. PMID- 16342107 TI - Calcium pyrophosphate deposition disease mimicking polymyalgia rheumatica: a prospective followup study of predictive factors for this condition in patients presenting with polymyalgia symptoms. AB - OBJECTIVE: To assess the characteristics of calcium pyrophosphate deposition disease (CPDD) with proximal involvement mimicking polymyalgia rheumatica (PMR), and to identify the best predictive factors for the presence of a clinical pattern of CPDD in patients presenting with polymyalgia symptoms. METHODS: Patients diagnosed with either PMR or CPDD at the Rheumatology Division of Hospital Meixoeiro (Vigo, Spain) over a 7-year period (1997-2003) were prospectively followed for at least 12 months. RESULTS: The study group comprised 118 patients with PMR features and 112 patients with CPDD. Eighty-two of the 118 patients with PMR manifestations were diagnosed as having pure PMR, and 36 met the diagnostic criteria for both PMR and CPDD. Patients with CPDD mimicking PMR were older (P = 0.02) and had peripheral arthritis more frequently (P = 0.004) than those with pure PMR. Radiologic osteoarthritic changes in the hands and knees, including more advanced radiologic grade of knee osteoarthritis, and tendinous calcifications were more frequent in patients with PMR/CPDD (P < 0.001). The best predictive factors for the occurrence of this atypical pattern of CPDD in a patient presenting with PMR features were the age at diagnosis and the presence of tibiofemoral osteoarthritis, tendinous calcifications, and ankle arthritis. CONCLUSION: Involvement of proximal joints may be the clinical presentation of CPDD. CPDD should be included in the spectrum of diseases mimicking PMR. The presence of tibiofemoral osteoarthritis, tendinous calcifications, and ankle arthritis are clues that may alert the clinician to the presence of CPDD in an elderly patient presenting with PMR manifestations. PMID- 16342108 TI - Preventing work disability among employees with rheumatoid arthritis: what medical professionals can learn from the patients' perspective. AB - OBJECTIVE: To compare the perspectives of employees with rheumatoid arthritis (RA) with those of medical professionals regarding what persons with RA need to prevent work disability. METHODS: Concept mapping was conducted in a group session with 21 employees and by mail with 17 medical professionals. Each group was asked to formulate statements on what enables employees with RA to retain their jobs. Group members scored all statements for importance and clustered them into themes. Results were statistically aggregated at the group level. RESULTS: The concept mapping with employees yielded 59 statements, which were clustered into 7 themes. The 4 most important themes were employer support; understanding and acceptance of illness by employees themselves; suitable working conditions; and support from colleagues, health professionals, and the patient's organization. The concept mapping with medical professionals yielded 65 statements, which were clustered into 8 themes. The 6 most important themes were well-informed professionals who cooperate effectively; employees' coping capacities and commitment to work; financial regulations at the workplace; adequate social security provisions, medication, and therapy; a positive attitude on the part of employers and colleagues; and suitable working conditions. CONCLUSION: Factors that enable continued employment lie at different levels, including the psychosocial, practical, organizational, and social policy levels. Health professionals appear to underestimate factors that are important from the patient's perspective, especially support from employers. In discussing work with patients, health professionals need to address themes that are important from the patient's perspective. PMID- 16342109 TI - Analyzing reliability of change in depression among persons with rheumatoid arthritis. AB - OBJECTIVE: To examine several methods of determining reliability of change constructs in depressive symptoms in patients with rheumatoid arthritis (RA) and to demonstrate the strengths, weaknesses, and uses of each method. METHODS: Data were analyzed from a cohort of 54 persons with RA who participated in a combined behavioral/pharmacologic intervention of 15 months duration. These longitudinal data were used to examine 3 methodologies for assessing the reliability of change for various measures of depression. The specific methodologies involved the calculations of reliable change, sensitivity to change, and reliability of the change score. RESULTS: The analyses demonstrated differences in reliability of change performance across the various depression measures, which suggest that no single measure of depression for persons with RA should be considered superior in all contexts. CONCLUSION: The findings highlight the value of utilizing reliability of change constructs when examining changes in depressive symptoms over time. PMID- 16342110 TI - Racial disparity in knee arthroplasty utilization in the veterans health administration. PMID- 16342112 TI - Laser scanning cytometer-based assays for measuring host cell attachment and invasion by the human pathogen Toxoplasma gondii. AB - BACKGROUND: Toxoplasma gondii is among the most common protozoan parasites of humans. Both attachment to and invasion of host cells by T. gondii are necessary for infection, yet little is known about the molecular mechanisms underlying these processes. T. gondii's etiological importance and its role as a model organism for studying invasion in related parasites necessitate a means to quantitatively assay host cell attachment and invasion. METHODS: We present here Laser Scanning Cytometer (LSC)-based assays of T. gondii invasion and attachment. The invasion assay involves automated counting of invaded and non-invaded parasites, differentially labeled with distinct fluorochromes. The attachment assay compares the relative binding of differentially labeled parasites. The assays were evaluated using treatments known to decrease invasion or attachment. RESULTS: The LSC-based assays are robust and reproducible, remove operator bias, and significantly increase the sample size that can be feasibly counted compared to other currently available microscope-based methods. In the first application of the new assays, we have shown that parasites attach to fixed and unfixed host cells using different mechanisms. CONCLUSIONS: The LSC-based assays represent useful new methods for quantitatively measuring attachment and invasion by T. gondii, and can be readily adapted to study similar processes in other host pathogen systems. PMID- 16342113 TI - Rho kinases regulate endothelial invasion and migration during valvuloseptal endocardial cushion tissue formation. AB - Rho-associated kinase (ROCK) is a downstream effector of small Rho-GTPases, and phosphorylates several substrates to regulate cell functions, including actin cytoskeletal reorganization and cellular motility. Endothelial-mesenchymal transformation (EMT) is a critical event in the formation of valves and septa during cardiogenesis. It has been reported that ROCK plays an important role in the regulation of endocardial cell differentiation and migration during mouse cardiogenesis (Zhao and Rivkees [2004] Dev. Biol. 275:183-191). Immunohistochemistry showed that, during chick cardiogenesis, ROCK1 and -2 were expressed in the transforming and migrating endothelial/mesenchymal cells in the outflow tract (OT) and atrioventricular (AV) canal regions from which valvuloseptal endocardial cushion tissue would later develop. Treatment with Y27632, a specific ROCK inhibitor, of cultured AV explants or AV endothelial monolayers of stage 14-minus heart (preactivated stage for EMT) on three dimensional collagen gel perturbed the seeding of mesenchymal cells into the gel lattice. In these experiments, Y27632 did not suppress the expression of an early transformation marker, smooth muscle alpha-actin. Moreover, Y27632 inhibited the mesenchymal invasion in stage 14-18 AV explants, in which endothelial cells had committed to undergo EMT. ML-9, a myosin light chain kinase inhibitor, also inhibited the mesenchymal invasion in cultured AV explants. These results suggest that ROCKs have a critical role in the mesenchymal cell invasion/migration that occurs at the late onset of EMT. PMID- 16342114 TI - Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure. AB - BACKGROUND: We characterize the application of multiphoton microscopy to the observation of the extracellular matrix of fresh unstained vessels. METHOD: Combined two-photon-excited fluorescence (2PEF) and second harmonic generation (SHG) imaging of large arteries reveals the architecture of elastin and collagen fibers in the vessel wall with remarkable specificity. RESULTS: We present elastin/collagen imaging in unstained rat vessels at both micrometer and whole vessel scales, and we characterize the optical properties of rat carotid artery and aorta walls. We apply this method to evidence deleterious effects of residual doses of a pesticide on the vessel wall. CONCLUSION: This study illustrates the potential of 2PEF/SHG microscopy for pharmacological studies in unlabeled arteries. PMID- 16342115 TI - Staining and quantification of poly-3-hydroxybutyrate in Saccharomyces cerevisiae and Cupriavidus necator cell populations using automated flow cytometry. AB - BACKGROUND: Poly [(R)-3-hydroxybutyric acid] (PHB) is a prokaryote storage material for carbon and energy that accumulates in cells under unbalanced growth conditions. Because this class of biopolymers has plastic-like properties, it has attracted considerable interest for biomedical applications and as a biodegradable commodity plastic. Current flow cytometric techniques to quantify intracellular PHB are based on Nile red. Here, an improved cytometric technique for cellular PHB quantification utilizing BODIPY 493/503 staining was developed. This technique was then automated using an automated flow cytometry system. MATERIALS: Using flow cytometry, the fluorescence of Saccharomyces cerevisiae and Cupriavidus necator with varying PHB content after staining with BODIPY 493/503 and Nile red was compared, and automated staining techniques were developed for both cultures. RESULTS: BODIPY 493/503 staining had less background staining, higher sensitivity and specificity to PHB, and higher saturation values than did Nile red staining. The developed automated staining procedure was capable of analyzing the PHB content of a bioreactor sample every 25 min and measured the average PHB content with accuracy comparable to offline GC analysis. CONCLUSION: BODIPY 493/503 produced an overall better staining for PHB than did Nile red. When combined with the automated system, this technique provides a new method for the online monitoring and control of bioreactors. PMID- 16342116 TI - Simultaneous quantitative measurement and automated analysis of mitochondrial morphology, mass, potential, and motility in living human skin fibroblasts. AB - BACKGROUND: Understanding the interdependence of mitochondrial and cellular functioning in health and disease requires detailed knowledge about the coupling between mitochondrial structure, motility, and function. Currently, no rapid approach is available for simultaneous quantification of these parameters in single living cells. METHODS: Human skin fibroblasts were pulse-loaded with the mitochondria-selective fluorescent cation rhodamine 123. Next, mitochondria were visualized using video-rate (30 Hz) confocal microscopy and real-time image averaging. To highlight the mitochondria, the acquired images were binarized using a novel image processing strategy. RESULTS: Our approach enabled rapid and simultaneous quantification of mitochondrial morphology, mass, potential, and motility. It was found that acute inhibition of mitochondrial complex I (NADH:ubiquinone oxidoreductase) by means of rotenone transiently reduced mitochondrial branching, area, and potential. In contrast, mitochondrial motility was permanently reduced. CONCLUSIONS: We present and validate a novel approach for rapid, unbiased, and simultaneous quantification of multiple mitochondrial parameters in living cells. Because this method is automated, large numbers of cells can be analyzed in a short period of time. PMID- 16342117 TI - Origin of the ultimobranchial body cyst: T/ebp/Nkx2.1 expression is required for development and fusion of the ultimobranchial body to the thyroid. AB - The ultimobranchial body (UBB) is an outpocketing of the fourth pharyngeal pouch that fuses with the thyroid diverticulum, giving rise to calcitonin-producing C cells. In this study, we demonstrate that the UBB is composed of two types of cells: one expressing T/ebp/Nkx2.1 and the other expressing p63. The former cell type, accounting for a majority of the UBB, requires T/ebp/Nkx2.1 for their survival. In contrast, the p63-positive cells, even in the absence of T/ebp/Nkx2.1 expression, can proliferate and give rise to a vesicular structure that is lined by a monolayer of p63-negative cells, surrounded by a cluster and/or single layer of p63-positive cells, displaying the basal/stem cell phenotype. T/ebp/Nkx2.1 haploinsufficiency causes abnormal fusion of the UBB with the thyroid diverticulum, which stays as a cluster of C-cells around the vesicular structure, similar to the one observed in mice null for T/ebp/Nkx2.1 expression. These results demonstrate that T/ebp/Nkx2.1 plays a role in the survival of UBB cells, their dissemination into the thyroid diverticulum, and the formation of UBB-derived vesicular structure. PMID- 16342118 TI - Erk1/2 signaling is required for Tgf-beta 2-induced suture closure. AB - Transforming growth factors beta (Tgf-betas) act by means of Smad signaling pathways and may also interact with the mitogen-activated protein kinase pathway. The hypothesis was tested that Erk1/2 signaling is required for Tgf-beta2-induced suture closure, by culturing embryonic mouse calvariae in the presence of Tgf beta2 with or without Erk1/2 inhibitor PD98059 (PD). Suture widths were measured daily, and microdissected sutures and bones were homogenized and protein analyzed by Western blots. Tgf-beta2 induced narrowing of the sutures after 72 hr, an effect inhibited by treatment with PD. Erk1/2 and Egf but not Smad2/3 protein expression was up-regulated by Tgf-beta2 calvarial tissues at 72 hr. PD inhibited endogenous and Tgf-beta2-stimulated Erk1/2 protein as well as Tgf-beta2 stimulated Egf, but increased Smad2/3 protein expression. In tissues harvested 0, 15, and 30 min after exposure to Tgf-beta2, Erk1/2 phosphorylation was up regulated after 15 min, an effect abrogated by the simultaneous addition of PD. In summary, Tgf-beta2 stimulated Erk1/2 phosphorylation and induced Egf and Erk1/2 expression, associated with suture closure after 72 hr. Blocking Erk1/2 activity with PD inhibited these effects but increased Smad2/3 expression. We postulate that Tgf-beta2 regulates suture closure directly by means of phosphorylation of Erk1/2 and indirectly by up-regulating Erk1/2, a substrate for Fgf receptor signaling required for Fgf induction of premature suture obliteration. PMID- 16342120 TI - Forebrain-specific knockout of B-raf kinase leads to deficits in hippocampal long term potentiation, learning, and memory. AB - Raf kinases are downstream effectors of Ras and upstream activators of the MEK ERK cascade. Ras and MEK-ERK signaling play roles in learning and memory (L&M) and neural plasticity, but the roles of Raf kinases in L&M and plasticity are unclear. Among Raf isoforms, B-raf is preferentially expressed in the brain. To determine whether B-raf has a role in synaptic plasticity and L&M, we used the Cre-LoxP gene targeting system to derive forebrain excitatory neuron B-raf knockout mice. This conditional knockout resulted in deficits in ERK activation and hippocampal long-term potentiation (LTP) and impairments in hippocampus dependent L&M, including spatial learning and contextual discrimination. Despite the widespread expression of B-raf, this mutation did not disrupt other forms of L&M, such as cued fear conditioning and conditioned taste aversion. Our findings demonstrate that B-raf plays a role in hippocampal ERK activation, synaptic plasticity, and L&M. PMID- 16342121 TI - Disease progression of human SOD1 (G93A) transgenic ALS model rats. AB - The recent development of a rat model of amyotrophic lateral sclerosis (ALS) in which the rats harbor a mutated human SOD1 (G93A) gene has greatly expanded the range of potential experiments, because the rats' large size permits biochemical analyses and therapeutic trials, such as the intrathecal injection of new drugs and stem cell transplantation. The precise nature of this disease model remains unclear. We described three disease phenotypes: the forelimb-, hindlimb-, and general-types. We also established a simple, non-invasive, and objective evaluation system using the body weight, inclined plane test, cage activity, automated motion analysis system (SCANET), and righting reflex. Moreover, we created a novel scale, the Motor score, which can be used with any phenotype and does not require special apparatuses. With these methods, we uniformly and quantitatively assessed the onset, progression, and disease duration, and clearly presented the variable clinical course of this model; disease progression after the onset was more aggressive in the forelimb-type than in the hindlimb-type. More importantly, the disease stages defined by our evaluation system correlated well with the loss of spinal motor neurons. In particular, the onset of muscle weakness coincided with the loss of approximately 50% of spinal motor neurons. This study should provide a valuable tool for future experiments to test potential ALS therapies. PMID- 16342122 TI - Inhibition of presynaptic activity by zinc released from mossy fiber terminals during tetanic stimulation. AB - Zinc exists in high densities in the giant boutons of hippocampal mossy fibers. On the basis of the evidence that zinc decreases extracellular glutamate concentration in the hippocampus, the presynaptic action of zinc released from mossy fibers during high-frequency (tetanic) stimulation was examined using hippocampal slices. The increase in zinc-specific fluorescent signals was observed in both extracellular and intracellular compartments in the mossy fiber terminals during the delivery of tetanic stimuli (100 Hz, 1 sec) to the dentate granule cell layer, suggesting that zinc released from mossy fibers is immediately retaken up by mossy fibers. When mossy fiber terminals were preferentially double-stained with zinc and calcium indicators and tetanic stimuli (100 Hz, 1 sec) were delivered to the dentate granule cell layer, the increase in calcium orange signal during the stimulation was enhanced in mossy fiber terminals by addition of CaEDTA, a membrane-impermeable zinc chelator, and was suppressed by addition of zinc. The decrease in FM4-64 signal (vesicular exocytosis) during tetanic stimulation (10 Hz, 180 sec), which induced mossy fiber long-term potentiation, was also enhanced in mossy fiber terminals by addition of CaEDTA and was suppressed by addition of zinc. The present study demonstrates that zinc released from mossy fibers may be a negative-feedback factor against presynaptic activity during tetanic stimulation. PMID- 16342123 TI - Applications of transition-metal catalysts to textile and wood-pulp bleaching. AB - From an economic perspective, textile and paper bleaching are amongst the most important oxidation processes. The removal of unwanted chromophores, be it stains on cloths or residual lignin in wood pulp, consumes more than 60 % of the world production of hydrogen peroxide. However, existing technologies have their limitations. At ambient temperature, hydrogen peroxide gives little stain bleaching and is used inefficiently. Hence the high product dosages and washing temperatures required limit its application to predominantly European markets, to the exclusion of the majority of the world's population. In paper manufacture, the use of chlorine-based oxidants results in the formation of chlorinated waste products, which show poor biodegradability. On the other hand, hydrogen peroxide requires higher temperatures, longer reaction times and is more expensive. Transition-metal catalysts offer an alternative. This review discusses the main classes of known bleach catalysts and their possible modes of action. PMID- 16342124 TI - Crown ether-gramicidin hybrid ion channels: dehydration-assisted ion selectivity. PMID- 16342125 TI - Mechanistic insight into the activity of tyrosinase from variable-temperature studies in an aqueous/organic solvent. AB - The activity of mushroom tyrosinase towards a representative series of phenolic and diphenolic substrates structurally related to tyrosine has been investigated in a mixed solvent of 34.4% methanol-glycerol (7:1, v/v) and 65.6% (v/v) aqueous 50 mM Hepes buffer at pH 6.8 at various temperatures. The kinetic activation parameters controlling the enzymatic reactions and the thermodynamic parameters associated with the process of substrate binding to the enzyme active species have been deduced from the temperature variation of the kcat and KM parameters. The activation free energy is dominated by the enthalpic term, the value of which lies in the relatively narrow range of 61+/-9 kJ mol(-1) irrespective of substrate or reaction type (monophenolase or diphenolase). The activation entropies are small and generally negative and contribute no more than 10% to the activation free energy. The substrate binding parameters are characterized by large and negative enthalpy and entropy contributions, which are typically dictated by polar protein-substrate interactions. The substrate 4 hydroxyphenylpropionic acid exhibits a strikingly anomalous temperature dependence of the enzymatic oxidation rate, with deltaH(double dagger) approximately = 150 kJ mol(-1) and deltaS(double dagger) approximately = 280 J K( 1) mol(-1), due to the fact that it can competitively bind to the enzyme through the phenol group, like the other substrates, or the carboxylate group, like carboxylic acid inhibitors. A kinetic model that takes into account the dual substrate/inhibitor nature of this compound enables rationalization of this anomalous behavior. PMID- 16342126 TI - In situ deposition and patterning of single-walled carbon nanotubes by laminar flow and controlled flocculation in microfluidic channels. PMID- 16342127 TI - The chemical synthesis of bioactive glycosylphosphatidylinositols from Trypanosoma cruzi containing an unsaturated fatty acid in the lipid. PMID- 16342129 TI - The discovery of novel metal-induced reactions of nitrones: not only electrophiles and reagents for [3+2] cycloadditions. PMID- 16342132 TI - Adolf von Baeyer: winner of the Nobel Prize for Chemistry 1905. PMID- 16342133 TI - Mechanism-based design of a ROMP catalyst for sequence-selective copolymerization. PMID- 16342134 TI - Total synthesis of (+)-rugulosin and (+)-2,2'-epi-cytoskyrin A through cascade reactions. PMID- 16342135 TI - Spontaneous oscillation and mechanically induced calcium waves in chondrocytes. AB - The characteristics of spontaneous calcium (Ca(2+)) oscillation and mechanically induced Ca(2+) waves in articular chondrocytes were studied. In some, but not all, chondrocytes in sliced cartilage and primary cultures, we observed spontaneous oscillation of intracellular Ca(2+) that never spread to adjacent cells. In contrast, a mechanical stimulus to a single cell by touching with a glass rod induced an increase of intracellular Ca(2+) that spread to neighboring cells in a wave-like manner, even though there was no physical contact between the cells. This indicated the release of some paracrine factor from the mechanically stimulated cells. Application of ultrasonic vibration also induced an oscillation of intracellular Ca(2+). The application of a uridine 5' triphosphate (UTP), UTP, induced a transient increase in intracellular Ca(2+) and the release of adenosine 5'-triphosphate (ATP) in cultured chondrocytes. A P2 receptor antagonist (suramin) and blockers of Cl(-) channels, niflumic acid and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), reduced the UTP-induced ATP release. The results indicated that Cl(-) channels were involved in the extracellular release of ATP following mechanical or P2Y receptor stimulation. Thus, ATP stimulation of P2Y receptors elicits an increase in intracellular Ca(2+), triggering further release of ATP from adjacent cells, thereby expanding the Ca(2+) wave in chondrocytes. PMID- 16342136 TI - The effect of 3-(5-nitro-2-thienyl)-9-chloro-5-morpholin-4-yl[1,2,4]triazolo[4,3 c]quinazoline on cell growth, cell cycle, induction of DNA fragmentation, and activity of caspase 3 in murine leukemia L1210 cells and fibroblast NIH-3T3 cells. AB - Quinazolines are multitarget agents, which have broad spectrum of biological activity, and some of them are now in cancer clinical testing. 3-(5-nitro-2 thienyl)-9-chloro-5-morpholin-4-yl[1,2,4]triazolo[4,3-c]quinazoline is a new synthetically prepared derivative, which in our previous study showed cytotoxic effects on cancer cell lines HeLa and B16. Quinazoline, at micromolar concentrations, induced morphological changes and necrosis of B16 cells, and at nanomolar concentrations it produced changes of F-actin cytoskeleton. It did not cause changes in the cell cycle, did not induce apoptotic cell death in B16 cells, did not have a mutagenic effect, and did not even behave as a typical intercalating agent. Little significant reduction of tumor volume in intramuscular transplanted B16 cells was observed. The aim of the present study was to examine the cytotoxic effect of 3-(5-nitro-2-thienyl)-9-chloro-5-morpholin 4-yl[1,2,4]triazolo[4,3-c]quinazoline on murine leukemia L1210 cells and fibroblast NIH-3T3 cells. Induction of cell morphology and cell cycle changes, induction of apoptosis and caspase 3 activity were studied. Quinazoline acted cytotoxically on both cell lines. The sensitivity of leukemia L1210 cells to the quinazoline was higher than that of fibroblast NIH-3T3. The IC(100) was 12 microM for L1210 cells and 24 microM for NIH-3T3 cells. No effect of quinazoline on the cell cycle profile of L1210 and NIH-3T3 was detected, however, quinazoline induced an increase of the sub-G(0) cell fraction, apoptotic DNA fragmentation, and apoptotic morphological changes at a concentration of 12 microM. This quinazoline concentration induced caspase 3 activity. Our results demonstrated that induction of apoptotic cell death via activation of caspase 3 contributed to the cytotoxic effects of 3-(5-nitro-2-thienyl)-9-chloro-5-morpholin-4 yl[1,2,4]triazolo[4,3-c]quinazoline in murine leukemia L1210 cells. PMID- 16342137 TI - Identification and characterization of a "biomarker of toxicity" from the proteome of the paralytic shellfish toxin-producing dinoflagellate Alexandrium tamarense (Dinophyceae). AB - The objective of this study was to identify and characterize a "biomarker of toxicity" from the proteome of Alexandrium tamarense, a paralytic shellfish toxin (PST)-producing dinoflagellate. A combination of 2-DE and MS approaches was employed to identify proteins of interest in the vegetative cells of several strains of A. tamarense with different toxin compositions and from different geographical locations. The electrophoretic analysis of the total water-soluble proteins from these toxic strains by 2-DE showed that several abundant proteins, namely AT-T1, AT-T2 and AT-T3, differing slightly in apparent Mr and pIs, were consistently present in all toxic strains of A. tamarense. Further analysis by MALDI-TOF MS and N-terminal amino acid sequencing revealed that they are isoforms of the same protein. Even more intriguing is that these proteins in A. tamarense have similar amino acid sequences and are closely related to a "biomarker of toxicity" previously reported in A. minutum. Unambiguous and highly species specific identification was later achieved by comparing the PMFs of proteins in these two species. An initial attempt to characterize these proteins by generation of murine polyclonal antibodies against the AT-T1 protein was successful. Western blot analysis using the murine AT-T1-polycolonal antibodies identified all the toxic strains of A. tamarense and A. minutum, but not the nontoxic strain of A. tamarense. These results indicate that these protein characteristics for toxic strains are species-specific and that they are stable properties of the tested algae which are clearly distinguishable irrespective of geographical location and toxin composition. To our knowledge, this is the first study to demonstrate the use of polyclonal antibodies against marker proteins purified from 2-DE gels to distinguish different strains and species of the PST producing dinoflagellate Alexandrium. It provides the basis for the production of monoclonal antibody probes against the "biomarkers of toxicity" for those dinoflagellates whose genome is incompletely characterized. Potentially, immunoassays could be developed to detect the presence of toxic algae in routine monitoring programs as well as to predict bloom development and movement. PMID- 16342138 TI - Quantitative proteome analysis of CD95 (Fas/Apo-1)-induced apoptosis by stable isotope labeling with amino acids in cell culture, 2-DE and MALDI-MS. AB - Proteome analysis of Jurkat T cells induced to undergo apoptosis by CD95 (Fas/Apo 1) treatment was performed to identify modified proteins. We used stable isotope labeling with amino acids in cell culture (SILAC) using leucine to identify proteins of apoptotic and control Jurkat T cells by 2-DE and MALDI-MS. Out of 224 spots analyzed, we quantified 213 spots with 3.5 leucine-containing peptide pairs on average; 28 proteins with a relative abundance of higher than 1.5 were found. Five new modified proteins including calcyclin binding protein, cytosolic acyl coenzyme A thioester hydrolase, heterogeneous ribonucleoprotein M, replication factor C 37-kDa subunit, and tropomyosin 4 chain were identified as being modified in response to apoptosis. In comparison to differential proteome analysis via silver-stained 2-D gels and PMF of total Jurkat T cell lysates, 15 additional apoptosis-modified proteins were identified though 8 proteins were not found. The described approach using SILAC instead of silver staining for relative quantification was simpler to perform regarding the number of required 2-D gels, that cumbersome gel comparisons were avoided, and more apoptosis-modified proteins were identified, but with a higher demand on data interpretation of the mass spectra obtained. PMID- 16342139 TI - Proteomic analysis of malignant lymphocyte membrane microparticles using double ionization coverage optimization. AB - Shed membrane microparticles (MPs) are microvesicles generated from the plasma membrane when cells are submitted to stress conditions. Although MPs reflect the cell state (at least in vitro), little is known on their protein composition. We describe the first set of experiments aiming to characterize the MP proteome. Two ways of triggering MP formation from a T-lymphocytic cell line were analyzed using a 1-D gel approach coupled with LC-MS/MS and the results were compared with those obtained from a classic membrane preparation. In total, 390 proteins were identified in MPs, among which 34% were localized to the plasma membrane. The MPs revealed a broad representation of plasma membrane proteins including 17 hematopoietic clusters of differentiation. This approach was successfully applied to one human chronic B-cell lymphoid malignancy. In all, 413 proteins were identified, including 117 membrane proteins, many of them being pathology associated. The sequence coverage in identified proteins was improved combining both nano-LC-MS/MS and MALDI-MS data. The suppression effect, observed on very complex peptide mixtures, was remediated by chromatographic fractionation. MPs may represent a new tool for studying plasma membrane proteins, displaying the advantages of reproducibility, minimal organelle contamination, and being potentially applicable to most cell types. PMID- 16342140 TI - Differential protein expression in the metal-reducing bacterium Geobacter sulfurreducens strain PCA grown with fumarate or ferric citrate. AB - Geobacter sulfurreducens, generally considered to be a strict anaerobe, is a predominant microbe in subsurface environments, where it utilizes available metals as electron acceptors. To better understand the metabolic processes involved in the metal-reduction capability of this microbe, the proteins expressed by cells grown anaerobically with either fumarate or ferric citrate as electron acceptor were compared. Proteins were separated by 2-DE under denaturing or nondenaturing conditions, and proteins varying in abundance with a high level of statistical significance (p<0.0001) were identified by peptide mass analysis. Denaturing 2-DE revealed significant differences in the relative abundance of the membrane proteins OmpA and peptidoglycan-associated lipoprotein, several metabolic enzymes, and, in addition, superoxide dismutase and rubredoxin oxidoreductase. Nondenaturing 2-DE revealed elevated catalase in cells grown with ferric citrate. These results suggest that, in addition to adjustments in membrane transport and specific metabolic pathways in response to these two different electron acceptors, distinct differences exist in the oxidative environment within the cell when fumarate or soluble ferric citrate is provided as electron acceptor. Although an anaerobe, G. sulfurreducens appears to have alternate mechanisms for dealing with reactive oxygen species in response to increased intracellular soluble iron. PMID- 16342141 TI - A robust meta-classification strategy for cancer detection from MS data. AB - We propose a novel method for phenotype identification involving a stringent noise analysis and filtering procedure followed by combining the results of several machine learning tools to produce a robust predictor. We illustrate our method on SELDI-TOF MS prostate cancer data (http://home.ccr.cancer.gov/ncifdaproteomics/ppatterns.asp). Our method identified 11 proteomic biomarkers and gave significantly improved predictions over previous analyses with these data. We were able to distinguish cancer from non-cancer cases with a sensitivity of 90.31% and a specificity of 98.81%. The proposed method can be generalized to multi-phenotype prediction and other types of data (e.g., microarray data). PMID- 16342142 TI - The proteome of mouse mucosal mast cell homologues: the role of transforming growth factor beta1. AB - Mast cells migrate to the mucosal epithelium during intestinal nematode infections in mice, where they express abundant mucosal mast cell-specific proteases, mouse mast cell protease-1 and -2 (MCPT1 and MCPT2). Expression of these proteases is strictly controlled by transforming growth factor-beta1 (TGF beta1) in the epithelium. In vitro homologues of mucosal mast cells are generated by culturing bone marrow-derived mast cells (BMMC) in the presence of TGF-beta1. We examined the proteome of BMMC cultured either in the presence of TGF-beta1 (n = 5) or of a neutralising anti-TGF-beta1 antibody (n = 5). Cell extracts were examined by 2-DE, and changes in expression levels of protein spots were determined by densitometry. Spots of interest were identified by tryptic peptide mapping. In addition to the up-regulation of MCPT1 and MCPT2, which accounted for approximately 40% of all soluble protein in the TGF-beta1 treated cells, MCPT7 was modestly up-regulated by TGF-beta1, and calnexin was up-regulated fivefold. A 7.6-fold down-regulation of galectin-1 was verified by Western blotting and FACS analysis. Galectin-1 is located on the cell surface where it mediates cellular adhesion to basement membranes. Regulation of its expression by TGF-beta1 may be of relevance to mast cell adhesion within the epithelium. PMID- 16342143 TI - Pedigree analysis of an elite rice hybrid using proteomic approach. AB - The definition of dominance or epistasis is generally on the basis of a descriptive characterization for these crops in the field, such as yield per hectare and the weight of grain. Since these trait examinations lack molecular information, how to precisely predict the phenotypic changes in filial generation is still a problem in heterosis studies. For rice, the genetic information caused by hybridization can be archived through analyzing of proteomes of rice seeds. Differential analysis of proteomes was introduced for the rice seeds of three cultivars, 9311, PA64S and LYP9, an elite rice hybrid from cross between 9311 and PA64S. In the three rice endosperms, the expression profiles of proteins were similar with the stained spots of 47 +/- 1, 46 +/- 0.6 and 44 +/- 0.6, for 9311, PA64S and LYP9, respectively; however, the number of proteins expressed in the rice embryos was significantly increased with the stained spots of 395.3 +/- 12.9, 350 +/- 9.2, and 389.3 +/- 16.4, for 9311, PA64S and LYP9, respectively. Importantly, the image comparisons and protein identifications have revealed in significantly different embryo protein spots among the three rice cultivars. By carefully analyzing these different 2-DE spots, many of them from the three embryos were shown to display a mirrored relationships between parents and the first filial generation. Furthermore, all of stained spots in LYP9 embryo were found on the 2-DEs from its parents, indicating that there was a genetic linkage. These results suggest that proteomic approach is able to serve pedigree analysis and functional prediction for new rice breeds. PMID- 16342144 TI - Microwave-assisted specific chemical digestion for rapid protein identification. AB - We have developed a rapid microwave-assisted protein digestion technique based on classic acid hydrolysis reaction with 2% formic acid solution. In this mild chemical environment, proteins are hydrolyzed to peptides, which can be directly analyzed by MALDI-MS or ESI-MS without prior sample purification. Dilute formic acid cleaves proteins specifically at the C-terminal of aspartyl (Asp) residues within 10 min of exposure to microwave irradiation. By adjusting the irradiation time, we found that the extent of protein fragmentation can be controlled, as shown by the single fragmentation of myoglobin at the C-terminal of any of the Asp residues. The efficacy and simplicity of this technique for protein identification are demonstrated by the peptide mass maps of in-gel digested myoglobin and BSA, as well as proteins isolated from Escherichia coli K12 cells. PMID- 16342145 TI - Functional characterization of the C-terminal domain of mouse capping enzyme. AB - Mouse capping enzyme (Mce1) consists of two functional domains: the amino terminal triphosphatase domain and the carboxyl-terminal guanylyltransferase (GTase) domain. The bifunctional Mce1 gene encodes 597 a.a. with a molecular weight approximately 68 kDa. Mce1 cDNA is located on chromosome 4A4 approximately 4A5 and is composed of 17 exons. To functionally characterize the C-terminus of Mce1, we generated four truncated proteins with 12, 30, 37, or 60 a.a. deletions from the C-terminus of either the wild type (Mce1) or the isolated GTase domain (211-597), respectively. Plasmid shuffling experiment with Saccharomyces cerevisiae GTase subunit gene CEG1 null mutant demonstrated that deletion mutants 211-567 and 211-585 were able to support cell viability in the presence of 5 fluoroorotic acid, whereas 211-537 and 211-560 were not. Consistent with the yeast genetic study, both 211-567 and 211-585 had significant GTase activity in vitro, while 211-537 and 211-560 that were only detected in the insoluble fraction in the bacterial expression system, were completely inactive. Overall, both in vivo and in vitro studies indicate that the functional domain of Mce1 is between a.a. 211 and 567, and the heptapeptide sequence between 561 and 567 may play an important role in the enzyme activity. PMID- 16342146 TI - Risk of second cancer among women with breast cancer. AB - A large number of women survive a diagnosis of breast cancer. Knowledge of their risk of developing a new primary cancer is important not only in relation to potential side effects of their cancer treatment, but also in relation to the possibility of shared etiology with other types of cancer. A cohort of 525,527 women with primary breast cancer was identified from 13 population-based cancer registries in Europe, Canada, Australia and Singapore, and followed for second primary cancers within the period 1943-2000. We used cancer incidence rates of first primary cancer for the calculation of standardized incidence ratios (SIRs) of second primary cancer. Risk of second primary breast cancer after various types of nonbreast cancer was also computed. For all second cancer sites combined, except contralateral breast cancer, we found a SIR of 1.25 (95% CI = 1.24-1.26) on the basis of 31,399 observed cases after first primary breast cancer. The overall risk increased with increasing time since breast cancer diagnosis and decreased by increasing age at breast cancer diagnosis. There were significant excesses of many different cancer sites; among these the excess was larger than 150 cases for stomach (SIR = 1.35), colorectal (SIR = 1.22), lung (SIR = 1.24), soft tissue sarcoma (SIR = 2.25), melanoma (SIR = 1.29), non melanoma skin (SIR = 1.58), endometrium (SIR = 1.52), ovary (SIR = 1.48), kidney (SIR = 1.27), thyroid gland (SIR = 1.62) and leukaemia (SIR = 1.52). The excess of cancer after a breast cancer diagnosis is likely to be explained by treatment for breast cancer and by shared genetic or environmental risk factors, although the general excess of cancer suggests that there may be additional explanations such as increased surveillance and general cancer susceptibility. PMID- 16342147 TI - Temporal diffusion spectroscopy: theory and implementation in restricted systems using oscillating gradients. AB - The theory of temporal diffusion spectra is reviewed. In contrast to q-space spectroscopy, which measures the displacement spectrum of spins in a spatial domain, the spectral density of the velocity correlation function (VCF) in the temporal domain is considered. It is demonstrated that casting diffusion in this domain may facilitate measurements of microscopic geometry and the decomposition of the diffusion signal into components due to disperse flow and restricted diffusion. An oscillating gradient (OG) method of diffusion spectroscopy was developed and implemented. Microscopic pore sizes, surface-to-volume ratios (S/Vs), and diffusion path tortuosities were extracted from model systems using this method. Cases are discussed in which this type of experiment may allow the characterization of pore geometry when spatial domain experiments fail. OGs may be combined with imaging sequences to map complex patterns of diffusion and flow. Moreover, scalar apparent diffusion coefficient (ADC) measurements in complex biological systems may be subtly dependent on specific pulse sequence parameters. Thus, scalar ADC measurements using gradient pulses with different frequency spectra may give different results. Conversely, the frequency dependence of motion-sensitizing gradient pulses may be exploited to deduce the origin of ADC changes. PMID- 16342148 TI - Gradient echo acquisition for superparamagnetic particles with positive contrast (GRASP): sequence characterization in membrane and glass superparamagnetic iron oxide phantoms at 1.5T and 3T. AB - Iron oxides are used for cell trafficking and identification of macrophages in plaque using MRI. Due to the negative contrast, differentiation between signal loss caused by iron and native low signal in tissue may be problematic. It is, therefore, preferable to achieve positive contrast. The purpose of this study was to test the efficacy of a new MRI sequence GRASP (GRe Acquisition for Superparamagnetic Particles) to generate a positive signal in phantoms containing iron. Membrane phantoms were constructed containing Ferumoxide at 7 concentrations. Standard GRE sequences were modified with user controlled z gradient rephasing (+/- 100%). CNR values were determined as a function of echo time (TE) and % rephasing at 1.5T and 3T. T(2)* values were determined using multiple double-echo GRE. The GRASP sequence generated positive signal enhancement in phantoms containing iron. For all rephasing values or = 3 in 4 patients; 31%; toxicity was graded according to the National Cancer Institute Common Toxicity Criteria [version 2.0]). Therapy was discontinued in one patient because of diarrhea not responding to dose adjustments. CONCLUSIONS: Single-agent lonafarnib appears to have clinical activity in a small proportion of patients with CML refractory to imatinib. PMID- 16342166 TI - Phase II study of epirubicin, cisplatin, and capecitabine for advanced biliary tract adenocarcinoma. AB - BACKGROUND: Advanced biliary tract carcinomas (BTCs) are associated with a very poor prognosis. New therapeutic strategies therefore are needed to improve efficacy and survival, and the current study was designed with a new, effective drug combination. METHODS: Patients with recurrent or metastatic BTC received a combination of epirubicin at a dose of 50 mg/m(2), cisplatin at a dose of 60 mg/m(2) on Day 1, and capecitabine at a dose of 1000 mg/m(2) twice daily for 2 weeks. Treatment was repeated every 3 weeks. RESULTS: A total of 43 patients (22 with extrahepatic cholangiocarcinoma, 15 with intrahepatic cholangiocarcinoma, and 6 with gallbladder carcinoma) were treated. The median age was 53 years (range, 36-69 yrs) and 5 patients had a Zubrod performance status of 2. Seventeen patients achieved a partial response (40%; 95% confidence interval [95% CI], 21 49%) and 10 had stable disease. With a follow-up duration of 18 months, the median survival time was 8 months (95% CI, 6-10 mos). In total, 187 chemotherapy cycles were delivered, with a median of 5 cycles per patient (range, 1-9 cycles). Toxicity was mainly myelosuppression and mucositis, but no patients died of toxicity. CONCLUSIONS: This combination chemotherapy with epirubicin, cisplatin, and capecitabine offered promising antitumor activity in patients with advanced BTC. PMID- 16342167 TI - Clinician to frontline soldier: a look at the roles and challenges of army clinical psychologists in Iraq. AB - Psychologists have a long and respected history of service during wartime that dates back to World War I. From the early days of scientific personnel selection, clinical psychologists have made a number of wartime contributions. Today, Army clinical psychologists serving in Iraq are providing behavioral health services as both clinicians and prevention specialists in order to meet the psychological and emotional needs of service members abroad. They are also performing as Army officers and soldiers contributing to the success of military operations as a whole. The authors, two clinical psychologists and Army officers currently deployed to Iraq, provide a small glimpse into the clinical and personal opportunities and challenges faced by wartime Army clinical psychologists. PMID- 16342168 TI - Construction of the perceived parental acculturation behaviors scale. AB - This article reports preliminary development of the Perceived Parental Acculturation Behaviors Scale (PPABS) based on a sample of 44 college students whose parents immigrated to America from Arab countries. The PPABS proposes two independent scales, whose contents characterize the respondents' perceptions of how much their parents evidence (a) openness to the American culture and (b) preservation of Arab culture. PMID- 16342169 TI - Immunocytochemical analysis of D-serine distribution in the mammalian brain reveals novel anatomical compartmentalizations in glia and neurons. AB - D-Serine is a co-agonist at the NMDA receptor glycine-binding site. Early studies have emphasized a glial localization for D-serine. However the nature of the glial cells has not been fully resolved, because previous D-serine antibodies needed glutaraldehyde-fixation, precluding co-localization with fixation sensitive antigens. We have raised a new D-serine antibody optimized for formaldehyde-fixation. Light and electron microscopic observations indicated that D-serine was concentrated into vesicle-like compartments in astrocytes and radial glial cells, rather than being distributed uniformly in the cytoplasm. In aged animals, patches of cortex and hippocampus were devoid of immunolabeling for D serine, suggesting that impaired glial modulation of forebrain glutamatergic signaling might occur. Dual immunofluorescence labeling for glutamate and D serine revealed D-serine in a subset of glutamatergic neurons, particularly in brainstem regions and in the olfactory bulbs. Microglia also contain D-serine. We suggest that some D-serine may be derived from the periphery. Collectively, our data suggest that the cellular compartmentation and distribution of D-serine may be more complex and extensive than previously thought and may have significant implications for our understanding of the role of D-serine in disease states including hypoxia and schizophrenia. PMID- 16342170 TI - 17beta-estradiol (betaE2) protects human retinal Muller cell against oxidative stress in vitro: evaluation of its effects on gene expression by cDNA microarray. AB - 17beta-estradiol (betaE(2)) is an effective neuroprotectant against hydrogen peroxide (H(2)O(2))-induced retinal neuronal cell death and light-induced photoreceptor degeneration. Muller cells are the principal macroglia responsible for supporting retinal neuronal survival, information processing and removing metabolic waste. However, the role of betaE(2) on human Muller cells is unclear. In this study, the effects of betaE(2) on human Muller cell survival and gene expression were examined. Our data revealed that betaE(2) is able to increase human Muller cell viability after exposure to H(2)O(2) through inhibition of apoptosis. Microarray analysis revealed significant changes in the expression of 69 genes (total of 21,324 genes screened) in cultured human Muller cells 6 h after betaE(2) treatment. Four of the betaE(2)-responsive genes [thrombospondin 1 (TSP1), mitogen-activated protein kinase kinase kinase 3 (MAP3K3), large conductance calcium-activated potassium channel beta2 subunit (KCNMB2), and SRY (sex-determining region Y)-box 11 (SOX11)] were validated by both real-time qRT PCR and semi-quantitative RT-PCR. Interestingly, exposure of human Muller cells to betaE(2) increased pigment epithelium-derived factor (PEDF) gene expression as measured by both RT-PCR and real time qRT-PCR. Our data demonstrate, for the first time, that betaE(2) protects cultured human Muller cells against H(2)O(2) induced cell death through the inhibition of apoptosis. This protective effect may operate through regulation of genes, such as TSP1, MAP3K3, SOX11, TSP1, and PEDF, and may in turn exert an important role in protecting retinal neurons. PMID- 16342171 TI - Macrophage/microglia activation factor expression is restricted to lesion associated microglial cells after brain trauma. AB - After traumatic brain lesion, microglial cells are rapidly activated, migrate toward the sites of injury, and cause secondary damage that accounts for most of the loss of brain function. In the present study, we have characterized a new macrophage/microglia activation factor (MAF). Using the monocytic cell line U937, we were able to demonstrate that MAF is upregulated after TPA-induced differentiation into macrophages. We have generated a specific antibody against MAF. In BV-2 microglial cells, MAF is partially co-localized with IB4, a classical microglial marker. In addition, we have analyzed the in vivo expression patterns of MAF after entorhinal cortex lesion. We were able to show a substantial upregulation of MAF on selected CD11b(+) and IB4(+) macrophages/microglial cells in the deafferented hippocampus and in the perilesional region, while no MAF expression was detectable on the contralateral side. Confocal microscopy revealed a lysosome-like expression pattern in BV-2 cells, as well as in ECL-associated macrophages/microglial cells in vivo. Furthermore, we were able to demonstrate that U937 cells with downregulated MAF converted slower and to a significantly reduced extent to the macrophageal phenotype after TPA treatment. In addition, MAF downregulation in BV-2 microglial cells substantially reduced the phagocytotic uptake of dextran beads. Our data indicate that MAF is expressed in selected macrophages/microglial cells around the lesion and in the degenerating hippocampus after ECL. Furthermore, MAF expression in monocytic cells seems to play a functional role in the differentiation to a phagocytosing phenotype and may be, at least partially, required for phagocytotic activity, specifically in lesioned tissue after brain trauma. PMID- 16342172 TI - EVI1 is consistently expressed as principal transcript in common and rare recurrent 3q26 rearrangements. AB - In contrast to the well-documented involvement of EVI1 in various 3q26 aberrations, the transcriptional status of EVI1 in rare recurrent or sporadic 3q26 chromosomal defects has remained largely unexplored. Moreover, in a recent report, the association between 3q26 alterations in myeloid proliferations and ectopic EVI1 expression was questioned. Therefore, we performed a detailed physical mapping of 3q26 breakpoints using a 1.3-Mb tiling path BAC contig covering the EVI1 locus and a carefully designed quantification of both EVI1 and MDS/EVI1 transcripts in 30 hematological malignancies displaying 3q26 aberrations. Cases included well-known rare, recurring chromosomal aberrations such as t(3;17)(q26;q22), t(2;3)(p21-22;q26), and t(3;6)(q26;q25), as well as 10 new sporadic cases. Extensive 3q26 breakpoint mapping allowed unequivocal and sensitive FISH detection of EVI1 rearrangements on both metaphases and interphase nuclei. Real-time quantitative PCR analyses indicated that typically both MDS1/EVI1 and EVI1, but not MDS1, were expressed in these malignancies, with EVI1 the primary transcript. In conclusion, we have demonstrated EVI1 involvement in numerous novel sporadic and recurrent 3q26 rearrangements. Our results underscore the feasibility of FISH as an adjunct to PCR for the identification of EVI1 deranged leukemias and identified EVI1 as the principal transcript expressed in these malignancies. PMID- 16342173 TI - Vascular hemichorea/hemiballismus and topiramate. PMID- 16342174 TI - Summary of contributions to GAW Group 5: linkage mapping methods, Problem 2. AB - Here I summarize the contributions to Group 5 of Genetic Analysis Workshop 14, held in Noordwijkerhout in The Netherlands. The theme of this group was linkage mapping methods applied to the simulated data (Problem 2). A variety of approaches were taken, and a number of questions were examined. In addition to testing for linkage in regions harboring known disease or modifying loci, or testing for linkage across the genome, the contributions addressed such issues as whether power/significance could be improved by making use of subphenotypes in addition to the primary disease phenotype, by modeling interactions between loci, or by using meta-analytic approaches to combine results from different populations. Most contributions were successful in identifying known disease loci D1 and D2, and for those contributions that examined the relevant regions, in identifying disease loci D3 and D4. The power to detect modifying loci D5 and D6 appeared to be lower. Some gain in power/significance was found from making use of subphenotypes and meta-analytic approaches. PMID- 16342175 TI - Haplotypes and haplotype-tagging single-nucleotide polymorphism: presentation Group 8 of Genetic Analysis Workshop 14. AB - Moderately dense maps of single-nucleotide polymorphism (SNP) markers across the human genome for both the simulated data set and data from the Collaborative Study of the Genetics of Alcoholism were available at Genetic Analysis Workshop 14 for the first time. This allowed examination of various novel and existing methods for haplotype analyses. Three contributors applied Mantel statistics in different ways for both linkage and association analysis by using the shared length between two haplotypes at a marker locus as a measure of genetic similarity. The results indicate that haplotype-sharing based on Mantel statistics can be a powerful approach and needs further methodological evaluation. Four contributors investigated haplotype-tagging SNP (htSNP) selection procedures, two contributors examined the use of multilocus haplotypes compared to single loci in association tests, and two contributors compared the accuracy of various methods for reconstructing haplotypes and estimating haplotype frequencies for both pedigree data and data from unrelated individuals. For all three different tasks, software packages and procedures gave similar results in regions of high linkage disequilibrium (LD). However, they were not as consistent in regions of moderate to low LD. One coalescence-based approach for estimating haplotype frequencies, coupled with a Markov chain Monte Carlo technique, outperformed the other haplotype frequency estimation methods in regions of low LD. In conclusion, regardless of the task, results were similar in chromosomal regions of high LD. However, based on the differing results observed here, methodological improvements are required for chromosomal regions of low to moderate LD. PMID- 16342176 TI - Summary of contributions to GAW Group 12: multivariate methods. AB - Here we summarize the contributions to Group 12 of Genetic Analysis Workshop (GAW) 14, held in Noordwijkerhout, The Netherlands. The theme of this group, multivariate methods, covered a broad range of statistical applications. Most of the contributors considered Problem 1 of the GAW. However, one paper considered the bivariate analysis of two binary phenotypes generated by the simulated data in Problem 2. Some contributors focused on statistical issues involved in considering multiple variables, and others on extensions to the variance components methodology for analysis of quantitative traits. Applications to the Collaborative Study on the Genetics of Alcoholism data identified a single nucleotide polymorphism (SNP) on chromosome 4 associated with the ttth1-ttth4 phenotypes, and replicated previous findings of linkage on chromosome 4 for alcohol consumption, using microsatellite and SNP data. PMID- 16342177 TI - Association mapping: methodologies, strategies, and issues. AB - Recent advances in molecular genetic technology allow for detailed characterization of genetic variation and easy cost-efficient accumulation of such data, even for large human samples. One such advance that presents incredible opportunities for identifying associations between genetic polymorphisms and disease-related phenotypes is the ability to quickly type a large number of single-nucleotide polymorphisms (SNPs). Contributors to Group 10 of Genetic Analysis Workshop 14 explored the potential of SNP genotypes for the association mapping of disease-related genes in family-based studies. Using both real data involving alcoholism susceptibility, made available by the Collaborative Study on the Genetics of Alcoholism (COGA), and simulated data involving personality-disorder susceptibility, group members investigated specific methodological issues involved in association mapping, such as multiple testing, single SNPs vs. combinations and haplotypes, and the effect of linkage disequilibrium on SNP-based linkage; evaluated existing methodologies for association mapping using SNPs, short-tandem repeats (STRs), or a combination of the two; and introduced new or modified association-mapping methods, including a gamma random effects (GRE) model and the quantitative trait linkage disequilibrium (QTLD) test. These papers are unified by the application of association-based methods to analyze SNPs, microsatellite markers, or both, to identify chromosomal regions harboring genes that contribute to quantitative endophenotype variation, and thus to disease risk. Their diversity attests to the breadth and flexibility of association-mapping approaches to the genetics of complex disease. PMID- 16342178 TI - Approaches to detecting gene x gene interaction in Genetic Analysis Workshop 14 pedigrees. AB - Whether driven by the general lack of success in finding single-gene contributions to complex disease, by increased knowledge about the potential involvement of specific biological interactions in complex disease, or by recent dramatic increases in computational power, a large number of approaches to detect locus x locus interactions were recently proposed and implemented. The six Genetic Analysis Workshop 14 (GAW14) papers summarized here each applied either existing or refined approaches with the goal of detecting gene x gene, or locus x locus, interactions in the GAW14 data. Five of six papers analyzed the simulated data; the other analyzed the Collaborative Study on the Genetics of Alcoholism data. The analytic strategies implemented for detecting interactions included multifactor dimensionality reduction, conditional linkage analysis, nonparametric linkage correlation, two-locus parametric linkage analysis, and a joint test of linkage and association. Overall, most of the groups found limited success in consistently detecting all of the simulated interactions due, in large part, to the nature of the generating model. PMID- 16342179 TI - Data mining. AB - Group 14 used data-mining strategies to evaluate a number of issues, including appropriate diagnosis, haplotype estimation, genetic linkage and association studies, and type I error. Methods ranged from exploratory analyses, to machine learning strategies (neural networks, supervised learning, and tree-based methods), to false discovery rate control of type I errors. The general motivations were to find the "story" in the data and to summarize information from a multitude of measures. Several methods illustrated strategies for better trait definition, using summarization of related traits. In the few studies that sought to identify genes for alcoholism, there was little agreement among the different strategies, likely reflecting the complexities of the disease. Nevertheless, Group 14 found that these methods offered strategies to gain a better understanding of the complex pathways by which disease develops. PMID- 16342180 TI - Genotyping errors, pedigree errors, and missing data. AB - Our group studied the effects of genotyping errors, pedigree errors, and missing data on a wide range of techniques, with a focus on the role of single-nucleotide polymorphisms (SNPs). Half of our group used simulated data, and half of our group used data from the Collaborative Study on the Genetics of Alcoholism (COGA). The simulated data had no missing genotypes and no genotyping errors, so our group, as a whole, removed data and introduced artificial errors to study the robustness of various techniques. Our teams showed that genotyping errors are less detectable and may have a greater impact on SNPs than on microsatellites, but recently developed methods that account for genotyping errors help reduce false positives, and the assumptions of these methods appear to be supported by observations from repeated genotyping. The ability to detect linkage disequilibrium (LD) was also substantially reduced by missing data; this in turn could affect tagging SNPs chosen to generate haplotypes. In the COGA sample, genotyping measurements were repeated in three ways. First, full-genome screens were performed on three sets of markers: 328 microsatellites, 11,560 SNPs from the Affymetrix GeneChip Mapping 10 K Array marker set, and 4,720 SNPs from the Illumina Linkage III panel. Second, the entire Affymetrix marker set was typed on the same 184 individuals by two different laboratories. Finally, the Affymetrix and Illumina marker panels had 94 SNPs in common. Our teams showed that both SNPs and microsatellites can be readily used to identify pedigree errors, and that SNPs have fewer genotyping errors and a low inconsistency rate. However, a fairly high rate of no-calls, especially for the Affymetrix platform, suggests that the inconsistency rate may be higher than observed. PMID- 16342182 TI - Linkage mapping methods applied to the COGA data set: presentation Group 4 of Genetic Analysis Workshop 14. AB - Presentation Group 4 participants analyzed the Collaborative Study on the Genetics of Alcoholism data provided for Genetic Analysis Workshop 14. This group examined various aspects of linkage analysis and related issues. Seven papers included linkage analyses, while the eighth calculated identity-by-descent (IBD) probabilities. Six papers analyzed linkage to an alcoholism phenotype: ALDX1 (four papers), ALDX2 (one paper), or a combination both (one paper). Methods used included Bayesian variable selection coupled with Haseman-Elston regression, recursive partitioning to identify phenotype and covariate groupings that interact with evidence for linkage, nonparametric linkage regression modeling, affected sib-pair linkage analysis with discordant sib-pair controls, simulation based homozygosity mapping in a single pedigree, and application of a propensity score to collapse covariates in a general conditional logistic model. Alcoholism linkage was found with > or =2 of these approaches on chromosomes 2, 4, 6, 7, 9, 14, and 21. The remaining linkage paper compared the utility of several single nucleotide polymorphism (SNP) and microsatellite marker maps for Monte Carlo Markov chain combined oligogenic segregation and linkage analysis, and analyzed one of the electrophysiological endophenotypes, ttth1, on chromosome 7. Linkage was found with all marker sets. The last paper compared the multipoint IBD information content of several SNP sets and the microsatellite set, and found that while all SNP sets examined contained more information than the microsatellite set, most of the information contained in the SNP sets was captured by a subset of the SNP markers with approximately 1-cM marker spacing. From these papers, we highlight three points: a 1-cM SNP map seems to capture most of the linkage information, so denser maps do not appear necessary; careful and appropriate use of covariates can aid linkage analysis; and sources of increased gene-sharing between relatives should be accounted for in analyses. PMID- 16342183 TI - Dissection of heterogeneous phenotypes for quantitative trait mapping. AB - We discuss analyses of Genetic Analysis Workshop 14 data from the Collaborative Study on the Genetics of Alcoholism (COGA) as well as from a simulated complex disease, Kofendrerd personality disorder (KPD), with both genetic and phenotypic heterogeneity. Both data sets included numerous related phenotypes in addition to disease definitions. All analyses either chose from the given selection of phenotypes or defined new ones, including traits that may not have been related to alcoholism or KPD. Some contributors evaluated the genetic components of the trait. Many investigated genome-wide linkage and/or association, using microsatellites and/or single-nucleotide polymorphism (SNP) chip data. Here we will focus on methodological issues that the investigators faced. Their results depended on phenotype selection, whether continuous or discrete, the covariates included, and ethnicity of the study population. For SNP chip data, members of our group detected no difference in results for Affymetrix or Illumina chips, although higher marker density for association studies appeared to be advantageous. Overall, there were some observations that different chromosomal segments, i.e., physical locations on the p-arm, q-arm, or middle segment, might lead to possible differences in type I error rates. This finding and others highlight the importance of empirical determination of P-values to determine significance. PMID- 16342184 TI - Fine mapping by linkage and association in nuclear family and case-control designs. AB - This report summarizes the Genetic Analysis Workshop 14 contributions related to fine-mapping strategies, in which examining smaller regions by association with single-nucleotide polymorphisms (SNPs) can yield savings in genotyping and multiple-testing penalties. The aim of the analyses conducted in Group 7 contributions was to localize disease susceptibility loci from either the simulated or the Collaborative Study on the Genetics of Alcoholism (COGA) data within identified regions of linkage. Among the 10 contributions, most groups analyzed the simulated data, one group analyzed the COGA data only, and one group analyzed both data sets. The research questions included evaluation of new methods of analysis, as well as comparisons among alternative methods, analytic strategies, and study designs. Methods of interest included an algorithm for SNP marker ordering, a locally weighted transmission disequilibrium test statistic, a likelihood-ratio test statistic for family-based association in nuclear families, a robust test statistic for case-control association studies, and Bayesian spatial modeling methods for haplotype clustering and association. Evaluations included comparisons among confidence intervals for loci detected via linkage, effects of multiple testing adjustments and trade-offs between type I error and power, comparisons among haplotype-based (multilocus) and genotype-based (multilocus and single-locus) association analyses, and design of fine-mapping and replication studies. While several promising new approaches were identified, further development and evaluation of methods for multiple testing, regression modeling of association with multiple markers and haplotypes, and combined treatment of linkage and association data are necessary if we are to identify many of the genes that contribute to complex traits. PMID- 16342185 TI - Assessment and implications of linkage disequilibrium in genome-wide single nucleotide polymorphism and microsatellite panels. AB - Linkage disequilibrium (LD) between markers is more likely to exist in dense genome-wide single-nucleotide polymorphism (SNP) panels than in microsatellite panels. As part of Genetic Analysis Workshop 14 (GAW14), the extent of LD in the Illumina linkage panel III and the Affymetrix Genechip 10 K mapping array was assessed, using data from the Collaborative Study on the Genetics of Alcoholism (COGA). The impact of LD on linkage results was examined in COGA and simulated data, and characteristics of SNPs were assessed for their ability to detect population substructure and predict haplotypes. The authors of the papers summarized here observed greater LD in the Affymetrix than in the Illumina panel, possibly due to increased marker density in the Affymetrix panel, and found greater LD on chromosome X than on the autosomes. Simulation analyses suggest that intermarker LD can cause an upward bias in linkage statistics; however, the impact of LD on linkage analysis depends on the proportion of ungenotyped founders and the extent of LD. No large effect of LD on linkage peaks was observed in COGA analyses. In addition, the papers summarized here found that SNPs with high minor allele frequencies were the most informative compared with microsatellites for the detection of population substructure, and that SNPs in higher LD, and small numbers of SNPs, were the most reliable for haplotype prediction. As ease of genotyping continues to increase, study design and SNP selection for linkage and association studies (including genome-wide association studies) will be improved with consideration of LD in the particular populations studied. PMID- 16342186 TI - Comparison of single-nucleotide polymorphisms and microsatellite markers for linkage analysis in the COGA and simulated data sets for Genetic Analysis Workshop 14: Presentation Groups 1, 2, and 3. AB - The papers in presentation groups 1-3 of Genetic Analysis Workshop 14 (GAW14) compared microsatellite (MS) markers and single-nucleotide polymorphism (SNP) markers for a variety of factors, using multiple methods in both data sets provided to GAW participants. Group 1 focused on data provided from the Collaborative Study on the Genetics of Alcoholism (COGA). Group 2 focused on data simulated for the workshop. Group 3 contained analyses of both data sets. Issues examined included: information content, signal strength, localization of the signal, use of haplotype blocks, population structure, power, type I error, control of type I error, the effect of linkage disequilibrium, and computational challenges. There were several broad resulting observations. 1) Information content was higher for dense SNP marker panels than for MS panels, and dense SNP markers sets appeared to provide slightly higher linkage scores and slightly higher power to detect linkage than MS markers. 2) Dense SNP panels also gave higher type I errors, suggesting that increased test thresholds may be needed to maintain the correct error rate. 3) Dense SNP panels provided better trait localization, but only in the COGA data, in which the MS markers were relatively loosely spaced. 4) The strength of linkage signals did not vary with the density of SNP panels, once the marker density was approximately 1 SNP/cM. 5) Analyses with SNPs were computationally challenging, and identified areas where improvements in analysis tools will be necessary to make analysis practical for widespread use. PMID- 16342187 TI - Case-control analyses: Geneopardy! AB - The case-control analyses group focused on a diverse set of questions, including study design, power, and analysis of case-control studies. Papers on study design examined various case-selection strategies, as well as ways to combine information from family- and population-based case-control data. The power of various approaches to case-control analyses was compared in several papers, while two papers chose to apply specific methods to analyzing the data. Two groups examined haplotype-related issues, including resolving un-phased haplotypes and comparing haplotype- and single-nucleotide polymorphism-based methods for analyzing data. Finally, one group examined the optimal number of markers for the genomic control approach to population stratification. PMID- 16342188 TI - Alcoholism and related traits: a summary of Group 13 contributions. AB - Ten groups set out to study the genetics of alcoholism, using various measures of alcohol dependence such as Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria, and related endophenotypes such as the electrophysiological evaluation of event-related potentials. The groups used both genome-wide microsatellite and single-nucleotide polymorphism (SNP) genotyping data in families selected from the Collaborative Study on the Genetics of Alcoholism. The majority of investigators studied alcohol-related phenotypes and chose linkage rather than association analysis. The analysis of SNP data presented several challenges, including marker linkage disequilibrium issues and computational limitations. Many groups pursued novel techniques, both in dealing with the SNP data and the definition of phenotypes. While there was a limited amount of concordance among linkage findings, it was very instructive to see so many new strategies at work. Generally the SNP genotype data seemed to yield more information for multipoint linkage analysis than the microsatellite data, a finding that will benefit the genetic analysis of complex disease in the future. A novel linkage peak was detected using the SNP markers. PMID- 16342189 TI - Heterogeneity: GAW Group 15. AB - This paper summarizes the contributions to Group 15 of Genetic Analysis Workshop (GAW) 14, which focused on methods for dealing with heterogeneity in linkage and association analysis. A variety of methods were employed, ranging from manipulation of the phenotype and/or identification of endophenotypes prior to analysis, to statistical methods allowing for heterogeneity in the analysis of simple dichotomous phenotypes as provided with the data. Overall, it was difficult to draw broad conclusions from these applications. Groups that focused on the simulated data had fairly consistent success in mapping the major genes (but not the two minor genes). However, this appeared to be the case regardless of whether heterogeneity was explicitly modeled, either at the phenotypic or statistical level, with relatively crude statistical methods applied to the simulated simple dichotomous trait Kofendrerd personality disorder also enjoying considerable success. PMID- 16342190 TI - Parent-of-origin, imprinting, mitochondrial, and X-linked effects in traits related to alcohol dependence: presentation Group 18 of Genetic Analysis Workshop 14. AB - The participants of Presentation Group 18 of Genetic Analysis Workshop 14 analyzed the Collaborative Study on the Genetics of Alcoholism data set to investigate sex-specific effects for phenotypes related to alcohol dependence. In particular, the participants looked at imprinting (which is also known as parent of-origin effect), differences between recombination fractions for the two sexes, and mitochondrial and X-chromosomal effects. Five of the seven groups employed newly developed or existing methods that take imprinting into account when testing for linkage, or test for imprinting itself. Single-marker and multipoint analyses were performed for microsatellite as well as single-nucleotide polymorphism markers, and several groups used a sex-specific genetic map in addition to a sex-averaged map. Evidence for paternal imprinting (i.e., maternal expression) was consistently obtained by at least two groups at genetic regions on chromosomes 10, 12, and 21 that possibly harbor genes responsible for alcoholism. Evidence for maternal imprinting (which is equivalent to paternal expression) was consistently found at a locus on chromosome 11. Two groups applied extensions of variance components analysis that model a mitochondrial or X-chromosomal effect to latent class variables and electrophysiological traits employed in the diagnosis of alcoholism. The analysis, without using genetic markers, revealed mitochondrial or X-chromosomal effects for several of these traits. Accounting for sex-specific environmental variances appeared to be crucial for the identification of an X-chromosomal factor. In linkage analysis using marker data, modeling a mitochondrial variance component increased the linkage signals obtained for autosomal loci. PMID- 16342191 TI - Heparin-induced thrombocytopenia type II: a rare but significant complication of plasma exchange. AB - Type II heparin-induced thrombocytopenia (HIT) is a rare but well-recognised and potentially life-threatening complication of unfractionated heparin therapy, and has been reported in association with heparin locks for central venous lines. We report a case of type II HIT complicated by iliofemoral deep venous thrombosis and pulmonary embolism in a 43-year-old woman in the course of plasma exchange for myasthenia gravis. A Gamcath central venous line had been inserted femorally due to poor peripheral venous access, and this was locked with heparin 5000 U/ml between procedures. Twelve days after initial heparin exposure, she presented with new-onset thrombocytopenia, a painfully swollen right leg, and pleuritic pain. Deep venous thrombosis and pulmonary embolism were confirmed radiologically, and serology for heparin/PF4 antibodies was unequivocally positive. The line was removed, and she was successfully managed with intravenous lepirudin, switching to warfarin on platelet recovery. This case demonstrates that Type II HIT can occur in association with heparin line locks in the course of plasmapheresis, despite previous reports of successful use of plasma exchange to treat Type II HIT. PMID- 16342192 TI - The acute phase reactant, fibrinogen, as a guide to plasma exchange therapy for acute Guillain-Barre syndrome. AB - The Guillian Barre syndrome is an acute inflammatory disorder for which plasma exchange is effective treatment. Up to 10% relapse after plasma exchange suggesting that treatment sometimes finishes before disease activity has resolved. We studied whether plasma fibrinogen, an inflammatory marker, might be used to determine when to discontinue plasma exchange in patients with acute Guillain-Barre syndrome. We conducted a post-hoc analysis of apheresis database and hospital records of patients treated with plasma exchange for acute Guillain Barre syndrome during 1999-2004. Data were analyzed from 28 patients who underwent a total of 29 courses of plasma exchange for acute Guillain-Barre syndrome. The mean (+/-SD) plasma fibrinogen concentration was 422.5 (+/-96.4) mg/dl at the time of presentation and, in 17 of the 29, it was above 400 mg/dl (reference range 200-400). Twenty of the 21 patients whose fibrinogen fell by more than 30% from baseline by the time of the final plasma exchange treatment had neurological improvement. There was improvement in only 3 of the 8 instances where fibrinogen decreased by less than 30% by the end of plasma exchange therapy. A > or =30% decrease in fibrinogen by the conclusion of plasma exchange was significantly associated with sustained neurological improvement (P = 0.0025). The plasma fibrinogen level appears to reflect disease activity in acute Guillain-Barre syndrome. A <30% fall in fibrinogen level despite plasma exchange may indicate the need to continue plasma exchange to maximize the benefit of treatment or minimize the risk of relapse. Therapeutic plasma exchange need not be extended when plasma fibrinogen remains > or =30% below its level at presentation by the time of the final planned plasma exchange procedure. PMID- 16342193 TI - Safety and efficacy of peripheral blood progenitor cell mobilization and collection in patients with advanced coronary heart disease. AB - Information on the safety of mobilization and collection of peripheral blood progenitor cells (PBPC) in patients with advanced coronary heart disease (CHD) is limited. We report herein our early experience with patients participating in a Phase I trial of injection of autologous CD 34(+) cells into threatened, ischemic myocardium for neovascularization and symptom relief in patients with chronic refractory myocardial ischemia. All patients had advanced inoperable CHD despite the best medical therapy. Granulocyte colony stimulating factor (G-CSF, 5 microg/kg/day) was administered subcutaneously for 5 days for mobilization of CD34(+) cells into the peripheral blood. PBPCs were collected in the outpatient apheresis suite on day 5. Nine patients from our institution were evaluable. Adverse effects of mobilization included: increase in frequency and/or intensity of angina in 8 patients (88.8%); bone pain in 7 patients (77.7%); headaches in 4 patients (44.4%); 2 patients (22%) were hospitalized. Collection phase toxicities included: tingling in 5 patients (55.5%) and angina in 3 patients (33%). All procedures were completed without new myocardial infarction, congestive heart failure, or death. The median peripheral blood CD34(+) cell count on day 5 of G CSF was 21 cells/microl (range 10-40 cells/microl). A median of 1.65 x 10(6) CD34(+) cells/kg (range: 0.13-3.0 x 10(6)/kg) were harvested. We conclude that mobilization and collection of PBPC in patients with advanced CHD can be safely performed as an outpatient procedure. Apheresis professionals should be aware of the intensity and frequency of angina in this patient population. PMID- 16342194 TI - Effects on apoptosis and reactive oxygen species formation by Jurkat cells exposed to 50 Hz electromagnetic fields. AB - The effect of exposure to 50 Hz electromagnetic field on a human T-leukaemia cell line (Jurkat) was investigated by evaluating the reactive oxygen species (ROS) production and apoptosis, both spontaneous and induced by a specific anti Fas/CD95 monoclonal antibody (anti-Fas). Our results suggest that 1 h intermittent (5 min field on/10 min field off) exposure does not affect ROS formation, while a slight but statistically significant decrease of both spontaneous and anti-Fas-induced apoptosis was observed. PMID- 16342195 TI - Effect of extremely low frequency electromagnetic fields (ELF-EMF) on Kaposi's sarcoma-associated herpes virus in BCBL-1 cells. AB - Association between extremely low frequency electromagnetic fields (ELF-EMF) and human cancers is controversial, and few studies have been conducted on their influence on oncogenic viruses. We studied the effects of 1 mT, 50 Hz sine waves, applied for 24-72 h, on Kaposi's sarcoma (KS)-associated herpesvirus (KSHV or HHV 8) in BCBL-1, a latently infected primary effusion lymphoma (PEL) cell line. ELF EMF exposure did not affect the growth and viability of BCBL-1 cells, either stimulated or not with TPA. The total amount of KSHV DNA detected in ELF-EMF exposed cultures not stimulated with TPA did not differ from that of the unexposed controls (P = ns). However, in the presence of TPA stimulation, total KSHV DNA content was found higher in ELF-EMF exposed than in control BCBL-1 cultures (P = .024) at 72 h exposure, but not earlier. Viral DNA increase significantly correlated with increased mean fluorescence intensity/cell for the lytic antigen gp K8.1A/B (P < .01), but not with percentage of gp K8.1A/B positive cells or of cells containing virions. Viral progeny produced under ELF EMF exposure consisted mainly of defective viral particles. PMID- 16342196 TI - A small temperature rise may contribute towards the apparent induction by microwaves of heat-shock gene expression in the nematode Caenorhabditis Elegans. AB - We have previously reported that low intensity microwave exposure (0.75-1.0 GHz CW at 0.5 W; SAR 4-40 mW/kg) can induce an apparently non-thermal heat-shock response in Caenorhabditis elegans worms carrying hsp16-1::reporter genes. Using matched copper TEM cells for both sham and exposed groups, we can detect only modest reporter induction in the latter exposed group (15-20% after 2.5 h at 26 degrees C, rising to approximately 50% after 20 h). Traceable calibration of our copper TEM cell by the National Physical Laboratory (NPL) reveals significant power loss within the cell (8.5% at 1.0 GHz), accompanied by slight heating of exposed samples (approximately 0.3 degrees C at 1.0 W). Thus, exposed samples are in fact slightly warmer (by < or =0.2 degrees C at 0.5 W) than sham controls. Following NPL recommendations, our TEM cell design was modified with the aim of reducing both power loss and consequent heating. In the modified silver-plated cell, power loss is only 1.5% at 1.0 GHz, and sample warming is reduced to approximately 0.15 degrees C at 1.0 W (i.e., < or =0.1 degrees C at 0.5 W). Under sham:sham conditions, there is no difference in reporter expression between the modified silver-plated TEM cell and an unmodified copper cell. However, worms exposed to microwaves (1.0 GHz and 0.5 W) in the silver-plated cell also show no detectable induction of reporter expression relative to sham controls in the copper cell. Thus, the 20% "microwave induction" observed using two copper cells may be caused by a small temperature difference between sham and exposed conditions. In worms incubated for 2.5 h at 26.0, 26.2, and 27.0 degrees C with no microwave field, there is a consistent and significant increase in reporter expression between 26.0 and 26.2 degrees C (by approximately 20% in each of the six independent runs), but paradoxically expression levels at 27.0 degrees C are similar to those seen at 26.0 degrees C. This surprising result is in line with other evidence pointing towards complex regulation of hsp16-1 gene expression across the sub-heat-shock range of 25-27.5 degrees C in C. elegans. We conclude that our original interpretation of a non-thermal effect of microwaves cannot be sustained; at least part of the explanation appears to be thermal. PMID- 16342197 TI - In vitro effects of GSM modulated radiofrequency fields on human immune cells. AB - Despite the important role of the immune system in defending the body against infections and cancer, only few investigations on possible effects of radiofrequency (RF) radiation on function of human immune cells have been undertaken. Aim of the present investigation was therefore to assess whether GSM modulated RF fields have adverse effects on the functional competence of human immune cells. Within the frame of the multidisciplinary project "Biological effects of high frequency electromagnetic fields (EMF)" sponsored by the National Occupation Hazard Insurance Association (AUVA) in vitro investigations were carried out on human blood cells. Exposure was performed at GSM Basic 1950 MHz, an SAR of 1 mW/g in an intermittent mode (5 min "ON", 10 min "OFF") and a maximum Delta T of 0.06 degrees C for the duration of 8 h. The following immune parameters were evaluated: (1) the intracellular production of interleukin-2 (IL 2) and interferon (INF) gamma in lymphocytes, and IL-1 and tumor necrosis factor (TNF)-alpha in monocytes were evaluated with monoclonal antibodies. (2) The activity of immune-relevant genes (IL 1-alpha and beta, IL-2, IL-2-receptor, IL 4, macrophage colony stimulating factor (MCSF)-receptor, TNF-alpha, TNF-alpha receptor) and housekeeping genes was analyzed with real time PCR. (3) The cytotoxicity of lymphokine activated killer cells (LAK cells) against a tumor cell line was determined in a flow cytometric test. For each parameter, blood samples of at least 15 donors were evaluated. No statistically significant effects of exposure were found and there is no indication that emissions from mobile phones are associated with adverse effects on the human immune system. PMID- 16342198 TI - Effect of short-wave (6-22 MHz) magnetic fields on sleep quality and melatonin cycle in humans: the Schwarzenburg shut-down study. AB - This paper describes the results of a unique "natural experiment" of the operation and cessation of a broadcast transmitter with its short-wave electromagnetic fields (6-22 MHz) on sleep quality and melatonin cycle in a general human population sample. In 1998, 54 volunteers (21 men, 33 women) were followed for 1 week each before and after shut-down of the short-wave radio transmitter at Schwarzenburg (Switzerland). Salivary melatonin was sampled five times a day and total daily excretion and acrophase were estimated using complex cosinor analysis. Sleep quality was recorded daily using a visual analogue scale. Before shut down, self-rated sleep quality was reduced by 3.9 units (95% CI: 1.7 6.0) per mA/m increase in magnetic field exposure. The corresponding decrease in melatonin excretion was 10% (95% CI: -32 to 20%). After shutdown, sleep quality improved by 1.7 units (95% CI: 0.1-3.4) per mA/m decrease in magnetic field exposure. Melatonin excretion increased by 15% (95% CI: -3 to 36%) compared to baseline values suggesting a rebound effect. Stratified analyses showed an exposure effect on melatonin excretion in poor sleepers (26% increase; 95% CI: 8 47%) but not in good sleepers. Change in sleep quality and melatonin excretion was related to the extent of magnetic field reduction after the transmitter's shut down in poor but not good sleepers. However, blinding of exposure was not possible in this observational study and this may have affected the outcome measurements in a direct or indirect (psychological) way. PMID- 16342199 TI - Gene expression profiles in Ciona intestinalis stigmatal cells: insight into formation of the ascidian branchial fissures. AB - Gill slits, a series of openings in the pharyngeal epithelium, are characteristic features of the hemichordate and chordate body plans. In ascidians, these openings, called stigmata, are formed in the branchial sac during juvenile development. Multiple whole-mount in situ hybridization analyses based on approximately 1,500 genes expressed in Ciona intestinalis juveniles, identified 28 genes expressed predominantly in the stigmatal cells. Expression patterns of these stigmatal genes were classified into four different categories. On the basis of these findings, we have been able to show that the peripheral region of a stigma consists of at least three different regions. The expression of a Dlk1 like gene was detected in nonciliated cells during the stigma perforation and division and was maintained in the basal region of the elliptical stigma. Expression of meichroacidin, tektin A1, and tektin B1 orthologs during the differentiation of the ciliated stigmatal cells suggests that some of the molecular mechanisms involved in sperm differentiation might be recruited for the stigma development, or vice versa. Components of the cilia such as alpha-tubulin and rootletin were also expressed in the stigmatal cells. These genes might facilitate further analyses regarding the evolution of the branchial fissures and the development of the ascidian stigmata. PMID- 16342200 TI - Reduction in diabetes-induced craniofacial defects by maternal immune stimulation. AB - BACKGROUND: Maternal diabetes can induce a number of developmental abnormalities in laboratory animals and humans, including facial deformities and defects in neural tube closure. The incidence of birth defects in newborns of diabetic women is approximately 3-5 times higher than among non-diabetics. In mice, non-specific activation of the maternal immune system can reduce fetal abnormalities caused by diverse etiologies, including diabetes induced neural tube defects. This study was conducted to determine whether non-specific maternal immune stimulation could reduce diabetes-induced craniofacial defects as well. METHODS: Maternal immune function was stimulated before streptozocin (STZ) treatment by maternal footpad injection with Freund's complete adjuvant (FCA), maternal intraperitoneal (i.p.) injection with granulocyte-macrophage colony-stimulating factor (GM-CSF), or maternal i.p. injection with interferon-gamma (IFNgamma). Streptozocin (200 mg/kg i.p.) was used to induce hyperglycemia (26-35 mmol blood glucose) in female ICR mice before breeding. Fetuses from 12-18 litters per treatment group, were collected at Day 17 of gestation. RESULTS: Craniofacial defects were observed in fetuses from all hyperglycemic groups. The incidence of defects was significantly decreased in fetuses from dams immune stimulated with IFNgamma or GM-CSF. The most common defects were reduced maxillary and mandibular lengths. Both were prevented by maternal stimulation with GM-CSF. CONCLUSION: Maternal immune stimulation reduced the incidence of diabetic craniofacial embryopathy. The mechanisms for these protective effects are unknown but may involve maternal or fetal production of cytokines or growth factors that protect the fetus from the dysregulatory effects of hyperglycemia. PMID- 16342201 TI - Complementary Gli activity mediates early patterning of the mouse visual system. AB - The Sonic hedgehog (Shh) signaling pathway plays a key role in the development of the vertebrate central nervous system, including the eye. This pathway is mediated by the Gli transcription factors (Gli1, Gli2, and Gli3) that differentially activate and repress the expression of specific downstream target genes. In this study, we investigated the roles of the three vertebrate Glis in mediating midline Shh signaling in early ocular development. We examined the ocular phenotypes of Shh and Gli combination mutant mouse embryos and monitored proximodistal and dorsoventral patterning by the expression of specific eye development regulatory genes using in situ hybridization. We show that midline Shh signaling relieves the repressor activity of Gli3 adjacent to the midline and then promotes eye pattern formation through the nonredundant activities of all three Gli proteins. Gli3, in particular, is required to specify the dorsal optic stalk and to define the boundary between the optic stalk and the optic cup. PMID- 16342202 TI - Two generation reproduction study of ethylbenzene by inhalation in Crl-CD rats. AB - BACKGROUND: This study was conducted to evaluate the potential adverse effects of ethylbenzene (EB) on reproductive capability from whole-body inhalation exposure of F0 and F1 parental animals. METHODS: Four groups of Crl:CD(SD)IGS BR rats (30/sex/group for F0 and 25/sex/group for F1) were exposed to 0, 25, 100, and 500 ppm EB for 6 hr/day for at least 70 consecutive days before mating. Inhalation exposure for the F0 and F1 females continued throughout mating, gestation through gestation day (GD) 20, and lactation days (LD) 5-21. On LD 1-4, females received EB in corn oil via oral gavage at dose levels of 26, 90, and 342 mg/kg/day (divided into three equal doses, approximately 2 hr apart), as calculated from a physiologically-based pharmacokinetic (PBPK) model to provide similar maternal blood area-under-concentration (AUC) as provided by inhalation. Pups were weaned on postnatal day (PND) 21 and exposure of the F1 generation started on PND 22. Estimates of internal exposure were determined by measuring EB concentrations in blood collected from F1 dams (4/group) and their culled pups 1 hr after the last gavage dose on PND 4. On PND 22, blood was collected from these same F1 dams and their weanlings for EB analysis 1 hr after a 6-hr inhalation exposure. The remainder of the F2 generation was not directly exposed. RESULTS: EB exposure did not affect survival or clinical observations. Male rats in the 500 ppm group in both generations gained weight more slowly than the controls. There were no indications of adverse effects on reproductive performance in either generation. Male and female mating and fertility indices, pre-coital intervals, spermatogenic endpoints, ovarian follicle counts, reproductive organ weights, lengths of estrous cycle and gestation, live litter size, pup weights, developmental landmarks, and postnatal survival were unaffected. No adverse exposure-related macroscopic pathology was noted at any level. CONCLUSIONS: Increased liver weights were found in the animals exposed to 500 ppm. F1 maternal whole blood EB concentrations of 0.49, 3.51, or 18.28 mg/L were found 1 hr after administration of a composite oral dose of 26, 90, or 342 mg/kg/day, respectively, but no detectable EB was found in blood samples of their F2 PND 4 culled pups. F1 maternal mean whole blood EB levels 1 hr after a 6-hr inhalation exposure on postpartum day (PPD) 22 was 0.11 mg/L (25 ppm), 0.56 mg/L (100 ppm), and 11 mg/L (500 ppm). For the offspring exposed with their dams on PND 22, F2 pup blood EB concentrations ranged from 0.017-0.039 mg/L (25 ppm), 0.165-0.465 mg/L (100 ppm), and 8.82-15.74 mg/L (500 ppm). Because decreased weight gain in the 500 ppm males was transient and no histopathological changes were associated with the increased liver weights in the 500 ppm male and female groups, these changes were not considered adverse. Therefore, for parental systemic toxicity, 100 ppm was considered a NOEL and 500 ppm a NOAEL in this study. The 500 ppm exposure concentration was considered a NOAEL for F0 and F1 reproductive toxicity and offspring developmental endpoints. PMID- 16342203 TI - Molecular characterization and expression of maternally expressed gene 3 (Meg3/Gtl2) RNA in the mouse inner ear. AB - The pathways responsible for sound perception in the cochlea involve the coordinated and regulated expression of hundreds of genes. By using microarray analysis, we identified several transcripts enriched in the inner ear, including the maternally expressed gene 3 (Meg3/Gtl2), an imprinted noncoding RNA. Real time PCR analysis demonstrated that Meg3/Gtl2 was highly expressed in the cochlea, brain, and eye. Molecular studies revealed the presence of several Meg3/Gtl2 RNA splice variants in the mouse cochlea, brain, and eye. In situ hybridizations showed intense Meg3/Gtl2 RNA staining in the nuclei of type I spiral ganglion cells and in cerebellum near the dorsal vestibular region of the cochlea. In embryonic mouse head sections, Meg3/Gtl2 RNA expression was observed in the otocyst, brain, eye, cartilage, connective tissue, and muscle. Meg3/Gtl2 RNA expression increased in the developing otocyst and localized to the spiral ganglion, stria vascularis, Reissner's membrane, and greater epithelial ridge (GER) in the cochlear duct. RT-PCR analysis performed on cell lines derived from the organ of Corti, representing neural, supporting, and hair cells, showed significantly elevated levels of Meg3/Gtl2 expression in differentiated neural cells. We propose that Meg3/Gtl2 RNA functions as a noncoding regulatory RNA in the inner ear and that it plays a role in pattern specification and differentiation of cells during otocyst development, as well as in the maintenance of a number of terminally differentiated cochlear cell types. PMID- 16342204 TI - Embryonic inner ear cells use migratory mechanisms to establish cell patterns in vitro. AB - The hair cells of the sensory epithelium in the inner ear are among the most precisely organized cells in vertebrates. The mechanisms that lead to this orderly arrangement are only beginning to be understood. It has been suggested that hair cells use migratory mechanisms to help achieve their final position in the organ of Corti. The small size and complex organization of the intact inner ear have made it difficult to monitor changes in hair cell location over time in vivo. In the present study, an established in vitro assay of dissociated, embryonic inner ear cells was used to monitor how hair cells reorganize over time. The hair cell specific marker myosin-VI demonstrated that hair cell precursors from both cochlear and vestibular regions reorganized into specific patterns between 3-24 hr in vitro. In contrast to the unlabeled cells, the myosin VI-positive cells extended processes while establishing the hair cell patterning within an aggregate. These studies support the hypothesis that hair cell precursors actively migrate to help achieve final patterning within the inner ear sensory epithelium. PMID- 16342205 TI - Alternative splicing in protein associated with Myc (Pam) influences its binding to c-Myc. AB - We recently identified Pam (for protein associated with c-Myc), as a binding partner for the tuberous sclerosis complex (TSC) protein tuberin in brain. The highly conserved Pam homologs in Drosophila and C. elegans are neuron-specific proteins that regulate synaptic growth. The Pam gene contains 83 exons and encodes a 4,641-amino-acid polypeptide with a predicted molecular weight of approximately 510 kDa. In a previous study, we demonstrated that Pam is expressed as two forms, approximately 450 kDa in rat embryonic and a approximately 350 kDa in rat adult brain. Here we have extended that work to show the approximately 450 kDa form is expressed in rat embryonic kidney, heart, and lung and in rat cell lines, and the approximately 350 kDa form is expressed in adult rat tissues as well as in human and mouse brain and human and mouse cell lines. To understand the size difference, we investigated alternative splicing of Pam in brain and detected six isoforms in the Myc-binding region resulting from splicing of exon 53, and three new exons, 52A, 56, and 56A. We also demonstrate that the presence of exon 52A in Pam significantly enhances binding to Myc, suggesting functional importance of this alternative splicing. The presence of Pam in many cellular compartments, its spliced variants, as well as its multiple binding partners, including tuberin, make it a complex, yet intriguing protein in the nervous system. PMID- 16342206 TI - Temporally specific proliferation events are induced in the hippocampus following acute focal injury. AB - In models of global brain injury, such as stroke or epilepsy, a large increase in neurogenesis occurs in the dentate gyrus (DG) days after the damage is induced. In contrast, more focal damage in the DG produces an increase in neurogenesis within 24 hr. To determine how cell proliferation and differentiation differs in the DG after acute injury in the DG, focal electrolytic lesions were made and mitotic activity was assessed at early (1 day) and late (5 day) time points. At the early time point, bromodeoxyuridine (BrdU)-positive cells were diffusely spread throughout the extent of the hippocampus that was ipsilateral to the lesion. No significant increase in the subgranular zone (SGZ) of the DG was observed. When BrdU was administered at the later time point, the number of BrdU+ cells in the SGZ of the DG was significantly increased. This fourfold increase in BrdU+ cells also resulted in a significant increase in neurogenesis, as measured 6 weeks following BrdU administration. This increase in neurogenesis was not observed when BrdU was administered at the early time point. These results indicate that focal injury in the DG activates two temporally specific proliferation events and that enhanced neurogenesis is observed only following a latent period after the lesion is made. PMID- 16342207 TI - In vivo delivery of fluoresceinated dextrans to the murine growth plate: imaging of three vascular routes by multiphoton microscopy. AB - Bone elongation by endochondral ossification occurs through the differentiation cascade of chondrocytes of cartilaginous growth plates. Molecules from the systemic vasculature reach the growth plate from three different directions: epiphyseal, metaphyseal, and a ring vessel and plexus associated with the perichondrium. This study is an analysis of the real-time dynamics of entrance of fluoresceinated tracers of different molecular weights into the growth plate from the systemic vasculature and tests the hypothesis that molecular weight is a key variable in the determination of both the directionality and the extent of tracer movement into the growth plate. Multiphoton microscopy was used for direct in vivo imaging of the murine proximal tibial growth plate in anesthetized 4- to 5 week-old transgenic mice with green fluorescent protein linked to the collagen II promoter. Mice were given an intracardiac injection of either fluorescein (332.3 Da) or fluoresceinated dextrans of 3, 10, 40, 70 kDa, singly or sequentially. For each tracer, directionality and rate of arrival, together with extent of movement within the growth plate, were imaged in real time. For small molecules (up to 10 kDa), vascular access from all three directions was observed and entrance was equally permissive from the metaphyseal and the epiphyseal sides. Within our detection limit (a few percent of vascular concentration), 40 kDa and larger dextrans did not enter. These results have implications both for understanding systemic and paracrine regulation of growth plate chondrocytic differentiation, as well as variables associated with effective drug delivery to growth plate chondrocytes. PMID- 16342209 TI - Developmental changes of nitric oxide synthase expression in the rat hypothalamoneurohypophyseal system. AB - The present study investigated the immunohistochemical localization of neuronal nitric oxide synthase (nNOS) in the hypothalamoneurohypophyseal system (HNS) of the developing rats on postnatal day 1 (PN1), 7 (PN7), 14 (PN14), 21 (PN21), and the adult rats. The nNOS-positive neurons were not discernable in the supraoptic nucleus (SON), the paraventricular nucleus (PVN), and the median eminence (ME) at PN1 and PN7. A few neurons positive for nNOS were first detected at PN14. At PN21, the nNOS-positive cells in SON and PVN rapidly increased in number. The pattern of nNOS expression at this stage approached that of the adult. Moreover, the increase of nNOS expression in the SON and PVN during the postnatal period was accompanied by the maturation of arginine vasopressin (AVP) and oxytocin (OT) neurons as indicated by the number and size of OT or AVP neurons in the SON and PVN. The patterns of AVP versus OT expression also reached that of the adult by the end of the third postnatal week. The time course of the change in nNOS expression coincided with the maturation of AVP and OT neurons in the HNS and suggested that NO synthesized by conversion of NOS is involved in the modulation of activity of neurons in the SON and PVN of the HNS. PMID- 16342208 TI - Molecular mimicry: sensitization of Lewis rats with Campylobacter jejuni lipopolysaccharides induces formation of antibody toward GD3 ganglioside. AB - Recently we have reported cases of demyelinating inflammatory neuropathy showing elevated titers of anti-GD3 antibodies, which occurs rarely in Guillain-Barre syndrome. To examine the correlation between the anti-GD3 antibody titer and Campylobacter jejuni infection, we sensitized female Lewis rats with lipopolysaccharides (LPSs) from serotype HS19 of C. jejuni and examined changes in nerve conduction velocity and nerve conduction block (P/D ratio). After 16 weeks of sensitization, animals revealed decreases of nerve conduction velocity and conduction block (P/D ratio) and high titer of anti-GD3 antibodies. These anti-GD3 antibodies also blocked transmission in neuromuscular junctions of spinal cord-muscle cells cocultures. The GD3 epitope was verified to be located on the Schwann cell surface and nodes of Ranvier in rat sciatic nerve. To determine the target epitope for GD3 antibodies in causing nerve dysfunction, the LPS fraction containing the GD3 epitope was purified from the total LPS by using an anti-GD3 monoclonal antibody-immobilized affinity column. Subsequently, chemical analysis of the oligosaccharide portion was performed and confirmed the presence of a GD3-like epitope as having the following tetrasaccharide structure: NeuAcalpha2-8NeuAc2-3Galbeta1-4Hep. Our data thus support the possibility of a contribution of GD3 mimicry as a potential pathogenic mechanism of peripheral nerve dysfunction. PMID- 16342210 TI - Postcranial myology of the California newt, Taricha torosa. AB - Salamanders are generally agreed to represent the primitive tetrapod body plan, as well as a postural analog for early tetrapods. Dissection and description of the muscles of the forelimb, trunk, and hindlimb of the California newt, Taricha torosa, were undertaken to provide baseline data on the locomotor structures in this species. Hypaxial trunk muscles are similar to those of other vertebrates. As in other salamanders, limb muscles show a simple parallel-fibered architecture and often span multiple joints. Several differences in limb musculature were also noted. The extensor iliotibialis muscle possesses a single head in T. torosa, rather than the two heads common in larger salamander species. The ischioflexorius muscle, while divided into proximal and distal sections, is not distinct from the puboischiotibialis in its proximal portion. The femorofibularis is enlarged in this species; it is suggested that the femorofibularis and ischioflexorius muscles may be functionally analogous systems. Forelimb and hindlimb musculature show similar morphological patterns, particularly in distal limb segments, which may provide insight into the primitive arrangement of tetrapod limb muscles. PMID- 16342211 TI - Effects of 12 days of artificial rearing on morphology of hypoglossal motoneurons innervating tongue retrusors in rat. AB - The purpose of this study was to examine the influence of reduced tongue activity by artificial rearing on the morphology of motoneurons innervating the extrinsic tongue retrusors. Artificially reared rat pups were fed via gastric cannula from postnatal day 3 to postnatal day 14. Artificially reared animals and dam-reared controls had cholera toxin (subunit B) conjugate of horseradish peroxidase injected into the styloglossus to label motoneurons innervating hyoglossus and styloglossus on postnatal day 13 and postnatal day 59. Following perfusion on postnatal days 14 and 60, serial transverse sections treated with tetramethyl benzidine and counterstained neutral red were used to analyze motoneuron morphology. The shorter diameter of hyoglossus motoneurons increased with age for the dam-reared but not the artificially reared group. There was a tendency for a similar pattern for styloglossus motoneurons across the two rearing groups. The changes in form factor reflected the changes in shorter diameter for both motoneuron pools. Therefore, reducing suckling activity during normal postnatal development leads to diminished motoneuron somal growth in rats. This may also be the case in premature infants necessarily fed artificially. PMID- 16342212 TI - Microstructure and innervation of the mystacial vibrissal follicle-sinus complex in bearded seals, Erignathus barbatus (Pinnipedia: Phocidae). AB - Vibrissal follicle-sinus complexes (F-SCs) are sensory receptors of the mammalian integument system. They are best developed within Pinnipedia. The objective of this study was to investigate the F-SCs of bearded seals (Erignathus barbatus) for benthic foraging adaptations. Bearded seals possessed approximately 244 mystacial F-SCs. In this species, F-SCs consisted of an outer dermal capsule (DC) surrounding a blood sinus system [upper cavernous sinus (UCS), ring sinus (RS), and lower cavernous sinus (LCS)] and concentric rings of epidermal tissue. The UCS comprised up to 62% of the F-SC length and may function as thermal protection for mechanoreceptors. A large asymmetrical ringwulst was located in the RS. A deep vibrissal nerve penetrated the DC at its base and terminated on mechanoreceptors in the epidermal tissues of the LCS and RS. The mean number of myelinated axons per F-SC was 1,314 (range, 811-1,650) and was among the highest number of axons per F-SC reported to date. An estimated mean number of 320,616 myelinated axons innervate the entire mystacial vibrissal array. Merkel-Neurite complexes (MNCs) and small simple laminated corpuscles were found in the region of the LCS. Myelinated axons also terminated on MNCs and lanceolate endings apical to the ringwulst. The number of F-SCs, their geometry in the mystacial region, the number of myelinated axons per F-SC, and the distribution of mechanoreceptors support the premise that pinniped vibrissae are sensitive active touch receptor systems, and that structural differences in bearded seals, relative to other phocids, may be adaptations for benthic foraging. PMID- 16342213 TI - Expression of caveolin-1 in the rat temporomandibular joint. AB - This immunocytochemical study revealed the expression of caveolin-1, a major protein of caveolae, in the rat temporomandibular joint. In the synovial lining layer, immunoreactive products for caveolin-1 were detected on the cell membrane of the fibroblast-like type B cells, as confirmed by immunocytochemistry for heat shock protein 25. The cells in the articular disk, the articular layer, and zone of proliferation of the mandibular condyle also showed intense immunoreactions for caveolin-1. PMID- 16342214 TI - Specializations of the granular prefrontal cortex of primates: implications for cognitive processing. AB - The biological underpinnings of human intelligence remain enigmatic. There remains the greatest confusion and controversy regarding mechanisms that enable humans to conceptualize, plan, and prioritize, and why they are set apart from other animals in their cognitive abilities. Here we demonstrate that the basic neuronal building block of the cerebral cortex, the pyramidal cell, is characterized by marked differences in structure among primate species. Moreover, comparison of the complexity of neuron structure with the size of the cortical area/region in which the cells are located revealed that trends in the granular prefrontal cortex (gPFC) were dramatically different to those in visual cortex. More specifically, pyramidal cells in the gPFC of humans had a disproportionately high number of spines. As neuron structure determines both its biophysical properties and connectivity, differences in the complexity in dendritic structure observed here endow neurons with different computational abilities. Furthermore, cortical circuits composed of neurons with distinguishable morphologies will likely be characterized by different functional capabilities. We propose that 1. circuitry in V1, V2, and gPFC within any given species differs in its functional capabilities and 2. there are dramatic differences in the functional capabilities of gPFC circuitry in different species, which are central to the different cognitive styles of primates. In particular, the highly branched, spinous neurons in the human gPFC may be a key component of human intelligence. PMID- 16342215 TI - Ossification of the mouse metatarsal: differentiation and proliferation in the presence/absence of a defined growth plate. AB - There is significant diversity in growth plate behavior among sites within an individual skeleton and between skeletons of different species. This variation within wild-type animals is an underutilized resource for studying skeletal development. One bone that potentially exhibits the most diverse behavior is the metatarsal. While one end forms a growth plate with an epiphyseal secondary center of ossification as in other long bones, the opposite end undergoes direct ossification in a manner more similar to short bones. Although descriptions of human metatarsal/metacarpal ossification are available, a detailed comparative analysis has yet to be conducted in an animal model amenable to biomolecular analysis. Here we report an analysis of proximal and distal ossification in an age series of mouse metatarsals. Safranin O staining was used for qualitative and quantitative histology, and chondrocyte differentiation and proliferation were analyzed using immunohistochemistry for type X collagen and proliferative cell nuclear antigen expression. We establish that, as in the human, both growth plate formation and direct ossification occur in the mouse metatarsal, with chondrocyte populations showing distinct differentiation patterns at opposite ends of the bone. In addition, growth plate formation is characterized by a peak of proliferation in reserve zone chondrocytes that distinguishes it from both established growth plates and direct ossification. Our analysis demonstrates that the mouse metatarsal is a productive model for investigating natural variation in ossification that can further understanding of vertebrate skeletal development and evolution. PMID- 16342216 TI - Sirolimus PK trial: a pharmacokinetic study of the sirolimus-eluting Bx velocity stent in patients with de novo coronary lesions. AB - This study was conducted to assess the systemic drug release and distribution of sirolimus-eluting stents. Early results with sirolimus-eluting stents have demonstrated a favorable outcome for reducing restenosis post coronary intervention. However, the clinical systemic pharmacokinetics of sirolimus released from these stents has not been investigated. Sirolimus-eluting stents (150-178 mcg/18 mm stent) were implanted in 19 patients with coronary artery disease using standard techniques. Blood samples were obtained at multiple times to determine the kinetics of sirolimus release and elimination. Non-compartmental analysis showed that the maximum blood concentration of sirolimus occurred between 3 and 4 hr after implantation, with a peak concentration of 0.57 +/- 0.12 ng/mL (mean +/- SD) and 1.05 +/- 0.39 ng/mL in patients receiving one or two stents, respectively. Terminal-phase elimination half-life was independent of the number of stents and averaged at 213 hr, a value longer than that seen in patients following oral dosing. The apparent clearance was 1.46 +/- 0.45 L/hr with an apparent volume of distribution in the terminal phase of 407 +/- 111 L (data for both stent doses pooled). Minimal measurable blood levels were detectable at 7 days. Peak whole blood level following sirolimus stent implantation in humans is proportional to the number of stents implanted. The prolonged terminal half-life may reflect kinetics of blood clearance combined with continued drug elution and secondary local tissue release. PMID- 16342217 TI - Aortic counterpulsation: a review of the hemodynamic effects and indications for use. AB - In the ever evolving field of cardiovascular medicine and coronary intervention, the intra-aortic balloon pump (IABP) is a mature technology, which still plays an important role. The balloon pump invasively supports patient hemodynamics by augmenting diastolic perfusion and increasing diastolic blood pressure, thereby increasing coronary perfusion and reducing afterload. Its efficacy has been demonstrated in a multitude of clinical situations, including acute coronary syndromes, high-risk coronary interventions, cardiogenic shock, and cardiovascular surgery. The potential complications of aortic counterpulsation are serious, although much lower than once feared. With proper patient selection, insertion technique, and management, the IABP is a powerful tool to assist in the treatment of patients with cardiovascular disease. This article will review the hemodynamics, indications, and complications associated with its use. PMID- 16342218 TI - Fine size control of platinum on carbon nanotubes: from single atoms to clusters. PMID- 16342219 TI - "Gating" the pores of a metal oxide based capsule: after initial cation uptake subsequent cations are found hydrated and supramolecularly fixed above the pores. PMID- 16342220 TI - Trimethylamine as a gas-phase promoter: highly efficient epoxidation of propylene over supported gold catalysts. PMID- 16342221 TI - The total synthesis of the fungal metabolite diversonol. PMID- 16342222 TI - An energetic measure of aromaticity and antiaromaticity based on the Pauling Wheland resonance energies. AB - Various criteria based on geometric, energetic, magnetic, and electronic properties are employed to delineate aromatic and antiaromatic systems. The recently proposed block-localized wave function (BLW) method evaluates the original Pauling-Wheland adiabatic resonance energy (ARE), defined as the energy difference between the real conjugated system and the corresponding virtual most stable resonance structure. The BLW-derived ARE of benzene is 57.5 kcal mol(-1) with the 6-311+G** basis set. Kistiakowsky's historical experimental evaluation of the stabilization energy of benzene (36 kcal mol(-1)), based on heats of hydrogenation, seriously underestimates this quantity due to the neglect of the partially counterbalancing hyperconjugative stabilization of cyclohexene, employed as the reference olefin (three times) in Kistiakowsky's evaluation. Based instead on the bond-separation-energy reaction involving ethene, which has no hyperconjugation, as well as methane and ethane, the experimental resonance energy of benzene is found to be 65.0 kcal mol(-1). We derived the "extra cyclic resonance energy" (ECRE) to characterize and measure the extra stabilization (aromaticity) of conjugated rings. ECRE is the difference between the AREs of a fully cyclically conjugated compound and an appropriate model with corresponding, but interrupted (acyclic) conjugation. Based on 1,3,5-hexatriene, which also has three double bonds, the ECRE of benzene is 36.7 kcal mol(-1), whereas based on 1,3,5,7-octatetraene, which has three diene conjugations, the ECRE of benzene is 25.7 kcal mol(-1). Computations on a series of aromatic, nonaromatic, and antiaromatic five-membered rings validate the BLW-computed resonance energies (ARE). ECRE data on the five-membered rings (derived from comparisons with acyclic models) correlate well with nucleus-independent chemical shift (NICS) and other quantitative aromaticity criteria. The ARE of cyclobutadiene is almost the same as butadiene but is 10.5 kcal mol(-1) less than 1,3,5-hexatriene, which also has two diene conjugations. The instability and high reactivity of cyclobutadiene thus mainly result from the sigma-frame strain and the pi-pi Pauli repulsion. PMID- 16342223 TI - A new efficient route to chiral 1,3-disubstituted ferrocenes: application to the syntheses of (R(p))- and (S(p))-17alpha-[(3'-formylferrocenyl)ethynyl]estradiol. AB - Starting from (2S,4S)-2-ferrocenyl-4-(methoxymethyl)-1,3-dioxane (4), use of the stereogenic ortho-directing menthyl para-tolyl sulfoxide group, which occupies the 2' position in the ferrocenyl ring and redirects subsequent lithiation to the 3' position, allowed the synthesis of optically pure (S(p))-1-formyl-3 iodoferrocene (8), that was characterized by single-crystal X-ray diffraction. Combination of this method with a protection-deprotection strategy, using trimethylsilyl as a temporary blocking group, yielded (R(p))-1-formyl-3 iodoferrocene (13). Separate Sonogashira coupling of each of the enantiomeric iodoformylferrocenes 8 and 13 with 17alpha-ethynyl-estradiol produced (R(p)) 17alpha-[(3'-formylferrocenyl)ethynyl]estradiol (14) and (S(p))-17alpha-[(3' formylferrocenyl)ethynyl]estradiol (15), respectively. PMID- 16342224 TI - Chemical waves. AB - In our paper we try to describe the basic concepts of chemical waves and spatial pattern formation in a simple way. We pay particular attention to self organisation phenomena in extended excitable systems. These result in the appearance of travelling waves, spiral waves, target patterns, Turing structures or more complicated structures called scroll waves, which are three-dimensional systems. We describe the most famous oscillating reaction, the Belousov Zhabotinsky (BZ) reaction, in greater detail. This is because it is of great interest in both physical chemistry and in studies on the evolution and sustenance of self-organising biological systems. PMID- 16342226 TI - Mechanism and exo-regioselectivity of organolanthanide-mediated intramolecular hydroamination/cyclization of 1,3-disubstituted aminoallenes: a computational study. AB - The complete catalytic reaction course for the organolanthanide-assisted intramolecular hydroamination/cyclization (IHC) of 4,5-heptadien-1-ylamine by a prototypical [(eta(5)-Me5C5)2LuCH(SiMe3)2] precatalyst has been critically scrutinized by employing a reliable DFT method. A computationally verified mechanistic scenario for the IHC of 1,3-disubstituted aminoallene substrates has been proposed that is consistent with the empirical rate law determined by experiment and accounts for crucial experimental observations. It involves kinetically rapid substrate association and dissociation equilibria, facile and reversible intramolecular allenic C=C insertion into the Ln-N bond, and turnover limiting protonation of the azacycle's tether functionality, with the amine amidoallene-Ln adduct complex representing the catalyst's resting state. This mechanistic scenario bears resemblance to the mechanism that has been recently proposed in a computational exploration of aminodiene IHC. The unique features of the IHC of the two substrate classes are discussed. Furthermore, the thermodynamic and kinetic factors that control the regio- and stereoselectivity of aminoallene IHC have been elucidated. These achievements have provided a deeper insight into the catalytic structure-reactivity relationships in organolanthanide-assisted cyclohydroamination of unsaturated C-C functionalities. PMID- 16342225 TI - Both spacer length and parity influence the thermal and light-induced properties of iron(II) alpha,omega-bis(tetrazole-1-yl)alkane coordination polymers. AB - A new series of [mu-tris-{1,n-bis(tetrazol-1-yl)alkane-N4,N4'}iron(II)] bis(perchlorate) spin-crossover coordination polymers ([Fe(nditz)3](ClO4)2]; n = 4-9) has been synthesised and characterised. The ditetrazole bridging ligands provide octahedral symmetry at the iron(II) centres while allowing the distance between iron(II) centres to be varied. These polymers have therefore been investigated to determine the effects of spacer length on their thermal and light induced spin-transition behaviour. An increase in the number of carbon atoms in the spacer (n) raises the thermal spin-crossover temperature, while decreasing the stability of the light-induced metastable state generated through the light induced excited spin state trapping (LIESST) effect by irradiating the sample at 530 nm. Remarkably, however, the parity of the spacer also has an effect, enabling the series of complexes to be divided into two sub-series depending on whether the bridging ligand possesses an even or an odd number of carbon atoms. An explanation at the molecular level using the single configurational coordinate (SCC) model is presented. PMID- 16342227 TI - A systematic review of the serotonergic effects of mirtazapine in humans: implications for its dual action status. AB - A systematic review of published work concerning mirtazapine was undertaken to assess possible evidence of serotonergic effects or serotonin toxicity (ST) in humans, because drug toxicity and interaction data from human over-doses is an useful source of information about the nature and potency of drug effects. There is a paucity of evidence for mirtazapine having effects on any indicator of serotonin elevation, which leads to an emphasis on ST as an important line of evidence. Mirtazapine is compared with its analogue mianserin, and other serotonergic drugs. Although mirtazapine is referred to as a dual-action 'noradrenergic and specific serotonergic drug' (NaSSA) little evidence to support that idea exists, except from initial microdialysis studies in animals showing small effects; those have not subsequently been replicated or substantiated by independent researchers. Also, new data indicate its affinity for Alpha 2 adrenoceptors is not different to mianserin. It appears to exhibit no serotonergic symptoms or toxicity in over-dose by itself, nor is there evidence that it precipitates ST in combination with monoamine oxidase inhibitors, as would be expected if it raises intra-synaptic serotonin levels. Mirtazapine has no demonstrable serotonergic effects in humans and there is insufficient evidence to designate it as a dual-action drug. PMID- 16342228 TI - In memoriam: Dr Terence Ivan Quickenden (1939-2005). PMID- 16342229 TI - The visualization of natural luminescence of living cotton hairs. AB - Super-weak luminescence of individual living cotton cells was investigated. This phenomenon was observed under an optical microscope using a replica-reprint technique in conjunction with living cotton cells. Gelatin solution was used for visualization of luminescence in a polymer film deposited on the cotton cells. No luminescence was generated in dead and very young plant cells (1 day after flowering) but maximal luminescence was detected from apical parts of cotton hairs in the early growth stage (4-6 days after flowering). Emitted luminescence had a cone-like form for different varieties of cotton plants. This indicates a focusing of radiation connected with morphological and structural features of the apical part of cotton cells at early stages of their evolution. The electromagnetic nature of cotton cell luminescence has been shown by experiments using photomultiplier (PEM) sensing of ultraviolet radiation. Insertion of a cotton seed-bud in darkness on a PEM window increased the dark current by 6-8%. Radiation flux from developing cotton hair is actually more intense, but through the PEM window only a small part of the radiation was detected. PMID- 16342231 TI - Biochemical profile in patients with anxious depression under the treatment with serotonergic antidepressants with different mechanisms of action. AB - The pharmacodynamics of serotonergic antidepressants that differentially influence serotonin reuptake transporters is poorly investigated. The aim of this study was to compare the biochemical profiles in patients with anxious depression under the treatment with tianeptine, a serotonin reuptake enhancer, and sertraline, a selective serotonin reuptake inhibitor. Platelet monoamine oxidase (MAO) and serum amine oxidase (AO) activities, concentration of middle-mass endotoxic molecules (MMEM) and parameters that characterize the functional properties of serum albumin were investigated in 43 patients with anxious depression (ICD-10: F 32.1 and F 33.1). It was established that, in comparison with healthy controls, patients with anxious depression were characterized by the significant increase in MAO activity (by 95%), MMEM concentration (by 86%), and a significant decrease in AO activity (by 43%) and also in functional albumin activity. The results of the study show that both tianeptine and sertraline are equally effective in the treatment of anxious depression. The present biochemical investigation, however, suggests that the underlying biochemical changes are more complete following tianeptine treatment. PMID- 16342232 TI - Expression of the NKG2D ligand UL16 binding protein-1 (ULBP-1) on dendritic cells. AB - Innate and adaptive immunity have not evolved separately. In this regard, the NKG2D molecule first identified on NK cells and classified as an activating NK cell receptor is also an important receptor for CD8(+) T cells. Functional analyses of human NKG2D and its ligands, i.e. UL16 binding proteins (ULBP) and MHC class I chain-related (MIC), have so far focused on immune cell-target cell situations because of the expression of NKG2D ligands on infected, stressed or transformed cells. Here, however, we address a possible function of NKG2D/ULBP-1 during the initiation of T cell responses. ULBP-1 can be detected on mature dendritic cells both in situ in the T cell areas of lymph nodes as well as in vitro after artificial maturation. FCM analysis further demonstrated that although NKG2D is expressed to some degree on all analyzed T cell subsets from peripheral blood, in vitro stimulation of T cells results in up-regulation of NKG2D on proliferating T cells. Using the sentinel lymph nodes of primary melanoma as a model for induction of defined T cell responses in vivo, we were able to demonstrate the expression of NKG2D on melanoma-associated antigen specific T cells. Thus, our results suggest a role for NGK2D-ULBP-1 in the induction or reactivation of T cell responses. PMID- 16342233 TI - Complex regulation of CCR9 at multiple discrete stages of T cell development. AB - We have conducted a comprehensive assessment of CCR9 expression and function at the important milestone stages of murine thymocyte development. We reveal an unusually complex regulatory pattern, in which CCR9 influences T cell development at several widely dispersed stages. We find that CCR9 is not expressed within the thymus until the double-negative (DN)3 stage, although it appears to contribute to T cell precursor development prior to residence in the thymus. CCR9 expression is influenced by pre-T cell receptor signals, and is dramatically up-regulated in a population that appears to be transitional between the DN4 and double-positive stages. In the periphery, functional CCR9 is expressed by all naive CD8 T cells, but not by naive CD4 T cells. To our knowledge, this latter finding is the first difference observed in homing receptor expression between naive lymphocyte populations. This suggests that naive CD8 T cells might have access to lymphoid microenvironments from which naive CD4 T cells are excluded. PMID- 16342234 TI - Induction of experimental autoimmune encephalomyelitis in transgenic mice expressing ovalbumin in oligodendrocytes. AB - We have used the 5' flanking sequence of the myelin basic protein gene known to include the core promoter and a strong oligodendrocyte (ODC)-specific enhancer to target expression of the well-studied model antigen ovalbumin (OVA) to ODC in transgenic mice. OVA protein was detected in a tissue- and cell-specific manner in these "ODC-OVA" mice. Without immunization, CD4 T cells and B cells remained ignorant of the neo-self antigen expressed in the central nervous system (CNS), as indicated by unimpaired development and lack of activation of OVA/IA(b) specific TCR transgenic T cells in these mice, and the ability to mount normal OVA-specific recall and antibody responses. Upon immunization with OVA in complete Freund's adjuvant, about half of the transgenic mice developed neurological symptoms characteristic of experimental autoimmune encephalomyelitis (EAE). Mononuclear infiltrates in the brain and spinal cord contained both macrophages and T cells, similar to classical models of EAE induced by immunization with CNS antigens in adjuvant. The wealth of immunological reagents available to study and manipulate the OVA-specific response should make this new model useful for the investigation of components and mechanisms involved in CNS specific autoimmunity. PMID- 16342235 TI - Preferential production of the IL-12(p40)/IL-23(p19) heterodimer by dendritic cells from human newborns. AB - Human newborns present impaired T helper type 1 cell responses, associated with a defect in the synthesis of IL-12 by dendritic cells (DC). IL-23 is a heterodimeric cytokine structurally related to IL-12, implicated in protective and autoimmune responses. We recently showed that upon activation neonatal T cells up-regulate a functional IL-23 receptor and that this cytokine polarizes the differentiation of naive T cells. We therefore investigated the capacity of neonatal DC to secrete IL-23. Lipopolysaccharide (LPS) stimulation induced the transcription of IL-23(p19) mRNA in both adult and neonatal DC, in sharp contrast to the repressed IL-12(p35) gene expression observed in neonatal cells. In comparison to adult DC, neonatal DC produced similar levels of IL-23 protein, in reponse to Toll-like receptor (TLR)-2- and TLR-3 ligands, and higher levels in response to TLR-4- or TLR-8 ligands. The same profile was observed in neonatal mononuclear cells. The supernatant of LPS-stimulated DC induced the secretion of IL-17 by polyclonally activated neonatal CD8(+) T cells, confirming the IL-23 bioactivity. Altogether, these observations strongly suggest that IL-23 could play a role in the immune system of human newborns. In particular, a functional IL-23/IL-17 axis might compensate a suboptimal IL-12/IFN-gamma pathway in early life. PMID- 16342236 TI - Inflammatory protein profile during systemic high dose interleukin-2 administration. AB - Systemic interleukin-2 (IL-2) administration induces an assortment of downstream effects whose biological and therapeutic significance remains unexplored mostly because of the methodological inability to globally address their complexity. Protein array analysis of sera from patients with renal cell carcinoma obtained prior and during high-dose IL-2 therapy had previously revealed extensive alterations in expression of the soluble factors analyzed, whose discovery was limited by the number of capture antibodies selected for protein detection. Here, we expanded the analysis to SELDI-TOF-MS and quantitative protein analysis (nephelometry). All cytokines/chemokines detected by protein arrays were below the SELDI detection limit, while novel IL-2-specific changes in expression of acute-phase reactants and high-density lipoprotein metabolites could be identified. Serum amyloid protein A (SAA) and C-reactive protein expression were consistently up-regulated after four doses of IL-2, while other proteins were down-regulated. These findings were confirmed by SELDI immunoaffinity capture and nephelometry. Immunoaffinity capture revealed different, otherwise undetectable, isoforms of SAA. A linear correlation between peak area by SELDI and protein concentration by nephelometry was observed. Overall distinct yet complementary information was obtained using different platforms, which may better illustrate complex phenomena such as the systemic response to biological response modifiers. PMID- 16342237 TI - Multidimensional proteomics of human serum using parallel chromatography of native constituents and microplate technology. AB - A versatile, multidimensional, and non-denaturing proteome separation procedure using microplate technology is presented, yielding a digitized image of proteome composition. In the first dimension, the sample under study is separated into 96 fractions by size exclusion chromatography (SEC). In the second dimension, the fractions of the first dimension are transferred by the liquid-handling device CyBi-Well (CyBio AG, Jena, Germany) to 96 parallel anion exchange chromatography columns. In this way the proteins are conserved in their native states and are distributed in 2400 liquid fractions with high recovery rates and sufficient reproducibility. The resulting fractions are subjected to protein quantitation and identification. Spectrophotometrical and immunological methods and enzyme activity measurements are used for quantitation. To identify proteins, the fractions are subjected to MALDI-MS, and their tryptic digests to both MALDI- and LC-ESI-MS/MS. All preparation steps except the first are applied in parallel to sets of multiples of 96 samples. The procedure may be refined by adding more separation steps and may be adapted to various protein amounts and to various proteomes. Moreover, the method offers the opportunity to investigate functional protein complexes. The method was applied to separate the normal human serum proteome. Within 255 fractions exhibiting the highest protein concentrations, 742 proteins were identified by LC-ESI-MS/MS peptide sequence tags. PMID- 16342238 TI - Detection of gender differences in rat lens proteins using 2-D-DIGE. AB - The glass-like transparency of the human eye lens is achieved by the tight packing of abundant crystallin proteins. However, the precise role of the accessory non-crystallin proteins is not well understood. We have carried out 2 DE mapping of these proteins in rat lens. This showed the presence of the high molecular weight filamentous structural proteins spectrin, filensin, tubulin, vimentin, actin and phakinin as well as several forms of potential crystallin oligomers comprised of alphaA, betaB1, betaA1 and betaA4 chains. Other proteins that were present include, heat shock protein 71, WD repeat protein 1, and several enzymes including alpha-enolase, pyruvate kinase, transketolase and aldose reductase. 2-D-DIGE analysis revealed several expression differences between the lens proteomes of male and female rats. Female rat lenses contained lower levels of aldose reductase, increased proteolyic fragments of the structural proteins filensin, vimentin and phakinin and higher levels of potential alphaA, betaB1 and betaA1 crystallin oligomers. Taken together these findings suggest that there are potential differences in oxidative stress regulation between male and female rat lenses, which may have implications on susceptibility to cataract formation. Future studies aimed at elucidating pre cataractic changes in the non-crystallin proteins described here may facilitate identification of novel markers involved in cataractogenesis. PMID- 16342239 TI - Phosphopeptide detection using automated online IMAC-capillary LC-ESI-MS/MS. AB - IMAC has become a commonly used technique in phosphoprotein analysis because of its affinity for phosphopeptides. However, the commonly used strategy combining offline IMAC enrichment with desalting procedures prior to MS/MS makes this method laborious. Here we report the development of a robust and automatic IMAC capillary RP HPLC-ESI MS/MS technology platform, by which all procedures needed in phosphopeptide analysis including IMAC enrichment, RP HPLC separation and nanospray MS/MS can be done automatically controlled by the MassLynx program. The platform was optimized by analyzing standard phosphopeptide, and was then applied to the identification of phosphorylation sites of recombinant human telomeric repeat binding factor 1 treated with kinase in vitro, and two phosphorylation sites are defined. PMID- 16342240 TI - Rapid protein identification using monolithic enzymatic microreactor and LC-ESI MS/MS. AB - A monolithic enzymatic microreactor was prepared in a fused-silica capillary by in situ polymerization of acrylamide, glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) in the presence of a binary porogenic mixture of dodecanol and cyclohexanol, followed by ammonia solution treatment, glutaraldehyde activation and trypsin modification. The choice of acrylamide as co-monomer was found useful to improve the efficiency of trypsin modification, thus, to increase the enzyme activity. The optimized microreactor offered very low back pressure, enabling the fast digestion of proteins flowing through the reactor. The performance of the monolithic microreactor was demonstrated with the digestion of cytochrome c at high flow rate. The digests were then characterized by CE and HPLC-MS/MS with the sequence coverage of 57.7%. The digestion efficiency was found over 230 times as high as that of the conventional method. In addition, for the first time, protein digestion carried out in a mixture of water and ACN was compared with the conventional aqueous reaction using MS/MS detection, and the former solution was found more compatible and more efficient for protein digestion. PMID- 16342241 TI - Proteome analysis of human lung squamous carcinoma. AB - Few lung cancer-specific molecular markers have been established in regard of "early-stage" diagnosis and prognosis. In this study the proteome analysis of human lung squamous carcinoma (hLSC) was carried out using two strategies to explore the carcinogenic mechanisms and identify its molecular markers more directly and comprehensively. Comparative proteome analysis on 20 hLSC tissues and paired normal bronchial epithelial tissues revealed 76 differential proteins, among which 68 proteins were identified by PMF. The identified proteins fell into three categories: oncoproteins, cell cycle regulators and signaling molecules. To validate the identified differential proteins, the expressions levels of three differential proteins mdm2, c-jun and EGFR were determined by immunohistochemical staining and immunoblots. The results verified proteome analysis results. Serological proteome analysis (SERPA) of ten hLSC tissues was performed to identify the tumor-associated antigens. The results revealed 36 +/- 8 differential proteins reactive with patients' autologous sera, of which 14 proteins were identified. Six of the 14 proteins, alpha enolase, pre-B cell enhancing factor precursor, triosephosphate isomerase, phosphoglycerate mutase 1, fructose-bisphosphate aldolase A, and guanine nucleotide-binding protein beta subunit-like protein, were also up-regulated in hLSCs in the comparative proteomic study, which suggests potential application of these 6 hLSC-associated antigens in diagnosis and therapy of hLSC. PMID- 16342242 TI - Proteome analysis of hepatocellular carcinoma by laser capture microdissection. AB - Hepatocellular carcinoma (HCC) is one of the most frequent visceral neoplasia worldwide and is a multifactorial and multistage pathogenesis that finally leads to the deregulation of cell homeostasis. Laser capture microdissection (LCM) may allow a more ready identification of differences in protein expression in selected cell types or areas of tissue, and microscopic regions as small as 3-5 microm in diameter can be sampled. Here we applied the LCM to the proteomic study of hepatitis B-related HCC and surrounding non-tumor tissues. Proteome alterations were observed using 2-DE and ESI-MS/MS, and alterations in the proteome were examined. Twenty protein spots were selected, of which 11 proteins were significantly altered in the HCC compared with the surrounding non-tumor tissues. Of the proteins that were selected, peroxiredoxin 2, apolipoprotein A-I precursor, 3-hydroxyacyl-CoA dehydrogenase type II, and 14.5-kDa translational inhibitor protein appear to be novel candidates as useful hepatitis B-related HCC markers. This study indicates that LCM is a useful technological method in the proteomic study of cancer tissue. The proteins revealed in this experiment can be used in the future for studies pertaining to hepatocarcinogenesis, or as diagnostic markers and therapeutic targets for HCC associated with hepatitis B virus infection. PMID- 16342243 TI - Use of detergents to increase selectivity of immunoprecipitation of tyrosine phosphorylated peptides prior to identification by MALDI quadrupole-TOF MS. AB - Identification of tyrosine phosphorylation by MS is challenging due to its low abundance in biological samples. Therefore, specific enrichment of tyrosine phosphorylated peptides prior to their analysis is highly desirable. The application of immunopurification of phosphotyrosine (pY) peptides using pY antibodies has been greatly limited by poor selectivity. In the present study, we have shown that the selectivity of pY peptide immunopurification can be dramatically improved by adding detergents to immunoprecipitation buffers. Optimum selectivity and sensitivity were achieved using an immunoprecipitation buffer containing n-octyl glucoside with a concentration above its critical micelle concentration (0.7%). The optimized method was used to identify in vivo tyrosine phosphorylation on proteins isolated from cell extract by anti-pY protein immunoprecipitation. After immunopurification, non-pY-containing peptides from protein digests were readily removed and pY peptides became the dominant peaks in MALDI quadrupole-TOF mass spectra. In addition, the signal intensities from pY-containing peptides were enhanced significantly after enrichment, allowing characterization of tyrosine phosphorylation sites with greater sensitivity. PMID- 16342244 TI - Proteomics-based generation and characterization of monoclonal antibodies against human liver mitochondrial proteins. AB - Monoclonal antibodies (mAbs) have the potential to be a very powerful tool in proteomics research to determine protein expression, quantification, localization and modification, as well as protein-protein interactions, especially when combined with microarray technology. Thus, a large amount of well-characterized and highly qualified antibodies are needed in proteomics. Purified antigen, which is not always available, has proven to be one of the rate-limiting steps in mAb large-scale generation. Here we describe our strategies to establish a murine hybridoma cell bank for human liver mitochondria using unknown native proteins as the immunogens. The antibody-recognized mitochondrial proteins were identified by MS following immunoprecipitation (IP), and by screening of human liver cDNA expression library. We found that the established antibodies reacted specifically with a number of important enzymes in mitochondria. The subcellular localization of these antigens in mitochondria was further confirmed by immunohistocytochemistry. A panel of antibodies was also tested for their ability to capture and deplete the targeting proteins and complexes from the total mitochondrial proteins. We believe these well-characterized antibodies would be useful in various applications for Human Liver Proteome Project (HLPP) when the scale of this hybridoma cell bank is enlarged significantly in the near future. PMID- 16342247 TI - Breast carcinoma during pregnancy. International recommendations from an expert meeting. AB - BACKGROUND: Breast carcinoma during pregnancy (BCP) is a difficult clinical situation, as it appears to put the health of the mother in conflict with that of the fetus. METHODS: An international expert meeting was conducted to form guidelines on how to diagnose and treat women with BCP. RESULTS: The goal for treatment of the pregnant woman with breast carcinoma is the same as that of the nonpregnant breast carcinoma patient: local control of disease and prevention of systemic metastases. However, certain treatment modalities need to be modified because of the potential for adverse effects on the fetus. There is evidence to support the safety of anthracycline-based chemotherapy during the second and third trimesters of pregnancy (Oxford Level of Evidence [LOE] 2b). Because of the lack of evidence, the expert opinion was not to recommend the routine use of newer cytotoxic drugs like the taxanes during pregnancy (LOE 5). CONCLUSION: The recommendations provided should help to reach informed decision making by the patient. The ongoing prospective collection of data on BCP, such as that at the University of Texas M.D. Anderson Cancer Center (UTMDACC) and that of the German Breast Group/Breast International Group (GBG/BIG), is necessary to further our knowledge regarding the treatment of this unique group of breast carcinoma patients. PMID- 16342248 TI - Detection of stool DNA mutations before and after treatment of colorectal neoplasia. AB - BACKGROUND: Whether stool DNA abnormalities arise solely from colorectal neoplastic lesions or are due to more pervasive field effects is not known. In the current study, the authors conducted a prospective multicenter study to evaluate the performance of stool-based DNA testing in a large cohort and to examine whether the findings before treatment persist after surgical resection and/or adjuvant therapy. METHODS: Patients with newly diagnosed colorectal carcinoma or advanced adenomas (AA) provided stool samples before therapy, 1-3 months after surgical resection, and 6-9 months postresection. Stool samples were analyzed using the multi-target DNA assay panel (MTAP) consisting of 23 markers: 21 mutations in the p53, K-ras, and APC genes, a microsatellite instability marker (BAT-26), and the DNA integrity assay (DIA), a marker of loss of apoptosis. RESULTS: Overall, 49 of 91 individuals (54%) tested positive with the MTAP test. The sensitivity of the MTAP test was 63% for invasive tumors compared with 26% for AA. Individuals whose lesions had a more advanced TNM stage or were located distal to the splenic flexure were significantly more likely to have a positive MTAP test. Of the 79 samples collected at 1-3 months after surgical resection of the neoplasm, 14 (18%) had a positive MTAP result, 12 of which were positive for DIA only. Of those collected at 6-9 months of follow-up, 5 of 72 (7%) tested positive on the MTAP panel. CONCLUSIONS: Although many samples collected 1-3 months after surgical resection of the colorectal neoplasm tested positive on the MTAP, most were negative by 6-9 months, indicating that stool DNA abnormalities disappear after treatment of the neoplastic lesions. Surgery and chemoradiation appear to induce transient DIA abnormalities that may be independent of the presence of neoplasia. PMID- 16342249 TI - Phase I studies of imatinib mesylate combined with cisplatin and irinotecan in patients with small cell lung carcinoma. AB - BACKGROUND: Small cell lung carcinoma (SCLC) cell lines commonly express KIT and its ligand, stem cell factor, suggesting an autocrine loop promoting cell growth. Imatinib inhibits KIT kinase activity. SCLC cells treated with imatinib in vitro undergo cell cycle arrest. Imatinib reduces resistance to irinotecan in vitro. Common metabolic pathways suggest there may be drug interactions between imatinib and irinotecan or cisplatin. In the current study, the authors investigated the feasibility of combining these drugs in the treatment of patients with SCLC. METHODS: Two Phase I studies were conducted independently at two institutions. Patients with extensive-disease SCLC underwent therapy with cisplatin, irinotecan, and imatinib using two similar regimens. In one study, immunohistochemical analysis of the expression of potential imatinib targets was performed on pretreatment biopsy specimens, and blood specimens were collected and analyzed for imatinib, irinotecan, and cisplatin pharmacokinetic parameters. RESULTS: Nine patients were enrolled and were evaluable for toxicity. A high incidence of neutropenia, diarrhea, and thrombosis was observed that precluded dose escalation. Six patients were evaluable for response after four cycles; five patients experienced a partial response and the other patient had developed progressive disease. Four of six tumor specimens tested expressed platelet derived growth factor receptor-alpha and two expressed KIT. Irinotecan clearance was found to be significantly decreased by imatinib (P < 0.04). No significant alteration in the disposition of cisplatin was observed. CONCLUSIONS: The maximum tolerated dose for this combination with granulocyte-colony-stimulating factor support was identified as imatinib at a dose of 300 mg/day with irinotecan (at a dose of 65 mg/m(2)) and cisplatin (at a dose of 30 mg/m(2)) given on Days 1 and 8, every 21 days. The decreased irinotecan clearance may explain the high incidence of diarrhea and neutropenia noted in the current study. PMID- 16342250 TI - Combination of paclitaxel and carboplatin as second-line therapy for patients with metastatic melanoma. AB - BACKGROUND: Patients with metastatic melanoma (MM) have very few therapy options. Based on reports of responses to paclitaxel and carboplatin (PC), 31 patients with MM were treated with PC. METHODS: Data regarding patients treated with PC were abstracted from medical records. Clinical outcomes as determined by the treating oncologist were used for this analysis. Response determination was retrospectively confirmed using Response Evaluation Criteria in Solid Tumors (RECIST). RESULTS: Thirty-one patients with MM were treated with PC. Patients had a median of 2 previous therapies, with the majority (29; 94%) having failed prior temozolomide (TMZ) or dacarbazine (DTIC) therapy. The most commonly used regimen was weekly paclitaxel (at a dose of 100 mg/m(2)) and carboplatin (area under the curve 2) administered on Days 1, 8, and 15 of a 28-day cycle. An objective partial response was noted in 8 patients (26%) with an additional 6 patients (19%) having stable disease; therefore, a clinical benefit was noted in 45% of those patients treated. The median time to disease progression for the entire group was 3 months (range, 0-7 mos), with a median overall survival of 7.8 months (range, 1-14 mos). The clinical benefit derived by the 14 patients, which lasted for a median of 5.7 months (range, 2.5-7.3 mos), was considered to be clinically significant. At the time of last follow-up, eight patients continued to receive PC therapy. CONCLUSIONS: The PC combination appears to have definite and clinically meaningful activity when used as second-line therapy after TMZ or DTIC. Further evaluation of this regimen, alone or as a 'backbone' for other agents, needs to be considered. PMID- 16342251 TI - Initial response to glucocorticoids. AB - BACKGROUND: Known prognostic variables in patients with primary central nervous system lymphomas (PCNSL) include age, Karnofsky performance status, involvement of deep regions of the brain, intensity of blood-brain barrier disruption, and treatment with radiation and chemotherapy. PCNSL often responds transiently to glucocorticoids administered to control neurologic symptoms before radiation or chemotherapy. This retrospective chart review was designed to estimate the prognostic significance of a clinical or radiologic response to initial glucocorticoid therapy. METHODS: By using data from The Johns Hopkins Cancer Registry from January 1980 to June 2001, a total of 76 human immunodeficiency virus (HIV)-negative adults with newly diagnosed PCNSL were identified. Nineteen patients with uninformative medical records were excluded from the study. RESULTS: The median survival of the remaining 57 patients was 11.8 months. The median survival for the 48 patients who had clinical response to initial steroid therapy was 17.9 months, and for nonresponders, it was 5.5 months (P = 0.05). The 16 patients with documented radiologic response had a median survival of 117.0 months compared with 5.5 months for nonresponders (P = 0.001). After adjusting for known prognostic factors (age and treatment), significant reduction in risk of death was noted in patients who had either clinical (hazard ratio [HR] = 0.40; 95% confidence interval [CI], 0.16-0.99}) or radiologic response (HR = 0.14; 95% CI, 0.04-0.46) to glucocorticoids given before radiation or chemotherapy. CONCLUSION: This analysis suggests that initial response to treatment with glucocorticoids may be an important prognostic factor in patients with PCNSL. PMID- 16342252 TI - Identification of relevant prognostic histopathologic features in 69 intracranial ependymomas, excluding myxopapillary ependymomas and subependymomas. AB - BACKGROUND: The results of attempts to identify histopathologic parameters that contribute to the clinical outcome of patients with ependymomas have been controversial. This may be due to the relative rareness of ependymomas. Furthermore, in many investigations, myxopapillary ependymomas and subependymomas were included and may have confounded results, because those tumors should be considered clinicopathologic entities distinct from the other ependymomas. METHODS: In this retrospective study, the influence of the histologic subtype of ependymoma and of individual histologic features on the outcome of 69 patients with ependymomas was investigated. Myxopapillary ependymomas, subependymomas, and ependymomas with spinal localizations were excluded from the analysis. The ependymomas were subdivided into cellular, papillary, clear cell, and tanycytic subtypes. The study extended over a period of 30 years. RESULTS: No differences in clinical outcome between the four histologic subtypes of ependymomas were revealed. Neither tumor localization (either infratentorial or supratentorial), patient age, nor gender affected survival. The survival of patients who underwent complete tumor resection differed significantly from that of patients who underwent partial resection. In univariate analysis, the features of nuclear atypia, the mitotic index, and the MIB-1 labeling index (LI) significantly influenced survival. With regard to survival, the presence of microcysts, blood vessel density, and the feature of vascular hyalinization demonstrated a trend but did not reach significance. In multivariate analysis, only the mitotic index and the MIB-1 LI were identified as factors with independent prognostic significance (P = 0.027 and P = 0.023, respectively). Both proliferation indices were correlated strongly with each other. CONCLUSIONS: The results of the univariate analysis indicated that, for patients with intracranial ependymoma, nuclear atypia, the mitotic index, and the MIB-1 LI significantly influenced survival. In the multivariate analysis, the mitotic index and the MIB-1 LI were the only features that had independent prognostic significance. Because both showed strong correlations, only one of them should be included in a grading scheme for intracranial ependymomas. PMID- 16342253 TI - Changing patterns of gallbladder carcinoma in New Mexico. AB - BACKGROUND: American Indians in the U.S. have a high incidence of gallbladder carcinoma (GBC). Furthermore, American Indians in New Mexico (NM) have the highest incidence rate of GBC in the U.S. The epidemiology of GBC in NM has not been studied in the past 3 decades. METHODS: By using the NM Tumor Registry (NMTR) and the Surveillance, Epidemiology, and End Results (SEER) database, age specific incidence rates, average annual age-adjusted incidence rates, and incidence rate time trends of GBC were compared among the three major ethnic groups in NM: American Indians, Hispanics, and non-Hispanic whites, for the period 1973-2001. RESULTS: A sharp increase in GBC incidence occurred with advanced age and started earlier in American Indians (at age 55) than Hispanics (at age 60) than non-Hispanic whites (at age 65). GBC occurred more frequently in females than in males in all ethnic groups. In females, the incidence of GBC was the highest in American Indians (14.5 per 100,000) followed by Hispanics (6.8 per 100,000) and non-Hispanic whites (1.4 per 100,000). Similarly in males, American Indians had the highest incidence rate of GBC (7.8 per 100,000), followed by Hispanics (2.0 per 100,000), and non-Hispanic whites (1.0 per 100,000). The time trend analysis revealed that there has been a disproportionate decline in the incidence of GBC in the three ethnic groups, with the greatest drop in American Indians and Hispanics followed by non-Hispanic whites. Despite the decline, American Indians continued to have the highest incidence rate of GBC in NM. CONCLUSION: The incidence of GBC has declined disproportionately in the three major ethnic groups, with the greatest decrease in American Indians and Hispanics followed by non-Hispanic whites. PMID- 16342254 TI - Local perspective of the impact of the HIPAA privacy rule on research. AB - BACKGROUND: The operational and economic impact of the Health Insurance Portability and Accountability Act (HIPAA) of 1996 was evaluated. The setting was a natural experiment which involved a single-site, clinical research study that was initiated before the enactment of HIPAA and subsequently modified to be compliant with the new policy. METHODS: A formative assessment was conducted of the recruitment process to a clinical trial evaluating the efficacy of an educational strategy to inform Veterans about the National Cancer Institute/Department of Veterans Affairs cosponsored Selenium and Vitamin E Cancer Prevention Trial (SELECT). Personnel time and costs were determined based on weekly accrual for study periods before and after the implementation of HIPAA. Root cause analysis was used to assess the recruitment protocol and to identify areas for improvement. RESULTS: The implementation of HIPAA resulted in a 72.9% decrease in patient accrual (7.0 patients/wk vs. 1.9 patients/wk, P < 0.001), and a threefold increase in mean personnel time spent recruiting (4.1 hrs/patient vs. 14.1 hrs/patient, P < 0.001) and mean recruitment costs (49 US dollars/patient vs. 169 US dollars/patient, P < 0.001). Upon review of the modified HIPAA compliant protocol, revisions in the recruitment procedure were adopted. The revised protocol improved weekly accrual by 73% (1.9 patients/wk vs. 7.1 patients/wk, P < 0.001) and resulted in improvements in personnel time (5.4 hrs/patient) and recruitment costs (65 US dollars/patient). CONCLUSION: Enactment of HIPAA initially placed a considerable burden on research time and costs. Establishing HIPAA-compliant recruitment policies can overcome some of these obstacles, although recruitment costs and time are likely to be greater than those observed before HIPAA. PMID- 16342255 TI - Globus pallidus deep brain stimulation in dystonia. AB - Globus pallidus deep brain stimulation (GPi-DBS) is a useful alternative in the treatment of dystonia. Patients selected for GPi-DBS were prospectively rated with the Unified Dystonia Rating Scale (UDRS). Also, "blinded" videotape assessments were performed. Eleven patients were identified. Compared with pre DBS scores, there were improvements in mean total UDRS score (15.3%) and in the following subscores: neck (18.18%), trunk (32.9%), arm (17.9%), and leg (19.9%). One patient developed a skin infection and erosion requiring surgical debridement. GPi-DBS is a safe and effective treatment for generalized dystonia in patients who remained impaired, despite optimal medical therapy. PMID- 16342256 TI - Progressive supranuclear palsy: phenotypic sex differences in a clinical cohort. AB - We examined sex-based differences in phenotypic expression among a consecutive clinical series of 121 individuals diagnosed with probable progressive supranuclear palsy (PSP). For both men (44%) and women (56%), the age at symptomatic onset (66.2 and 68.5 years, respectively) and disease duration (4.6 and 4.3 years, respectively) were similar. The overwhelming majority of sex-based comparisons showed no significant difference on a variety of demographic, historical, and clinical characteristics, as well as measures of disease progression. The only differences observed were that men had significantly worse tremor as measured by the Unified Parkinson's Disease Rating Scale tremor subscore (0.9 for men and 0.3 for women, P<0.01) and men had a significantly higher mean body mass index (BMI; 28.2 for men and 25.1 for women, P=0.01), although these differences were not significant after Bonferroni correction. In general, the disease phenotype was similar between men and women, suggesting that sex may have little or no influence on the development, expression, or progression of the PSP phenotype. PMID- 16342257 TI - Function of snake mobbing in spectral tarsiers. AB - Numerous species are known for their tendency to approach and confront their predators as a group. This behavior is known as mobbing. Snakes seem to be one of the more consistent recipients of this type of predator-directed behavior. This paper explores individual differences (sex and age) in the mobbing behavior of the spectral tarsier toward live and model snakes. This study was conducted at Tangkoko Nature Reserve (Sulawesi, Indonesia) during 2003-2004. During this research, 11 natural mobbing events and 31 artificially induced mobbing events were observed. The mean number of individuals at a mobbing was 5.7. The duration of mobbing events was strongly correlated with the number of assembled mobbers. Adults were more likely than other age classes to participate in mobbings. Males were more likely than females to participate in mobbings. Mobbing groups often contained more than one adult male, despite the fact that no spectral tarsier group contains more than one adult male. No difference in body size between extragroup males and resident males was observed, refuting the "attract the mightier" hypothesis. The number of mobbers did not affect whether the tarsier or the snake retreated first, countering the "move-on" hypothesis. The "perception advertisement" hypothesis was tentatively supported, in that live snakes were rarely seen in the area following mobbing calls, in comparison to when tarsiers either ignored the snake or alarm call. PMID- 16342258 TI - Molecular characterization of a pre-Columbian mummy and in situ coprolite. AB - The history of Homo sapiens dispersal around the world and inherent interpopulation contacts and conflicts has given rise to several transitions in his relationships with the natural world, with the final result of changes in the patterns of infectious disease (McMichael [2001] Ecosystem Health 7:107-115). Of particular interest, in this context, is the contact between Amerindians and Europeans that started at the end of the 15th century, and the resulting exchange of microbes. We successfully recovered ancient DNA from a pre-Columbian mummy from Cuzco (Peru), radiocarbon-dated to 980-1170 AD, for which consistent mtDNA amplifications and sequences were obtained. The analysis of mtDNA revealed that the mummy's haplogroup was characteristic of Native American populations. We also investigated a sample of feces directly isolated from the intestines of the mummy, using a polymerase chain reaction system designed to detect the broadest spectrum of bacterial DNAs. The analysis of results, following a criterion of "paleoecological consistency" (Rollo and Marota [1998] Philos. Trans. R. Soc. Lond. [Biol.] 354: 111-119), demonstrated that some vestiges of the original microbial flora of the feces were preserved. In particular, we were able to identify the DNA of Haemophylus parainfluenzae, thus suggesting that this recently recognized pathogen was present in precontact Native Americans. PMID- 16342259 TI - Is human longevity a consequence of cultural change or modern biology? AB - Increased longevity, expressed as the number of individuals surviving to older adulthood, represents a key way that Upper Paleolithic Europeans differ from earlier European (Neandertal) populations. Here, we address whether longevity increased as a result of cultural/adaptive change in Upper Paleolithic Europe, or whether it was introduced to Europe as a part of modern human biology. We compare the ratio of older to younger adults (OY ratio) in an early modern human sample associated with the Middle Paleolithic from Western Asia with OY ratios of European Upper Paleolithic moderns and penecontemporary Neandertals from the same region. We also compare these Neandertals to European Neandertals. The difference between the OY ratios of modern humans of the Middle and Upper Paleolithic is large and significant, but there is no significant difference between the Neandertals and early modern humans of Western Asia. Longevity for the West Asian Neandertals is significantly more common than for the European Neandertals. We conclude that the increase in adult survivorship associated with the Upper Paleolithic is not a biological attribute of modern humans, but reflects important cultural adaptations promoting the demographic and material representations of modernity. PMID- 16342260 TI - Age-related cortical bone loss in women from a 3rd-4th century AD population from England. AB - Age-dependent cortical bone loss in adult females from a skeletal assemblage from 3rd-4th century AD England was studied using metacarpal radiogrammetry. Results showed reduced peak cortical bone thickness compared with modern subjects, and the magnitude of cortical bone loss in older females compared with their younger counterparts was greater than that documented for a modern reference population. An elevated prevalence of fractures classically associated with osteoporosis was also observed in the over-50-year cohort. The severity of osteoporosis in this group is difficult to explain in terms of extraneous factors relating to 3rd-4th century lifestyles. Given the important genetic component in osteoporosis, the results may indicate some inherent susceptibility in this particular population to the disease, and ways in which this possibility might be further explored are suggested. PMID- 16342261 TI - Transmission of migration propensity increases genetic divergence between populations. AB - The advent of molecular genetics has brought invaluable information, which is now routinely used by anthropologists in their attempt to reconstruct our demographic past. Since mitochondrial DNA loci are much more similar between populations than are Y-chromosome loci, it is suggested that women had a much higher migration rate than men throughout history. Based on an examination of intergenerational migration patterns in three large demographic databases, we bring this inference into question. In some early Canadian settlements (St. Lawrence Valley and Saguenay), and in the former Krummhorn region of northwest Germany, men whose fathers were migrants were more likely to migrate, while the migration probability of women was largely independent of that of their mothers. As a result, men's movements were less effective in preventing genetic differentiation between populations than women's movements. In order to account for the impact of transmission, we propose a slight modification of Wright's island model. We also address the relevance of this model at the regional scale, and we discuss the supporting historical and anthropological literature. We conclude that the widespread patrilocal rules of postmarital residence have generated both a higher female migration rate and a patrilineal dependency in the propensity to migrate. PMID- 16342262 TI - Alteration of protein subcellular location and domain formation by alternative translational initiation. AB - Alternative translation is an important cellular mechanism contributing to the generation of proteins and the diversity of protein functions. Instead of studying individual cases, we systematically analyzed the alteration of protein subcellular location and domain formation by alternative translational initiation in eukaryotes. The results revealed that 85.7% of alternative translation events generated biological diversity, attributed to different subcellular localizations and distinct domain contents in alternative isoforms. Analysis of isoelectric point values revealed that most N-terminal truncated isoforms significantly lowered their isoelectric point values targeted at different subcellular localizations, whereas they had conserved domain contents the same as the full length isoforms. Furthermore, Fisher's exact test indicated that the two ways targeting at different cellular compartments and changing domain contents-were negatively associated. The N-term truncated isoforms should have only one way to diversify their functions distinct from the full-length ones. The peculiar consequence of subcellular relocation as well as change of domain contents reflected the very high level of biological complexity as alternative usage of initiation codons. PMID- 16342263 TI - Steady state of gradient echo sequences with radiofrequency phase cycling: analytical solution, contrast enhancement with partial spoiling. AB - Spoiled gradient echo sequences can only reach a homogeneous steady state if sufficiently strong crusher gradients are used in combination with RF phase cycling (RF spoiling). However, the signal depends quite sensitively on the chosen phase increment phi and-lacking analytical solutions-numerical simulations must be used to study the transient and steady-state magnetization. For the steady state an exact analytical solution is derived, which holds for arbitrary sequence and tissue parameters. Besides a considerably improved computation performance, the analytical approach enables a better understanding of the complicated dependence on phi. For short repetition times (TR) the regime of small phi turns out to be particularly interesting: It is shown that the typical phi(c), where RF spoiling starts to become effective, is essentially inversely proportional to T(2). This tissue dependence implies that contrasts can be considerably larger with partial spoiling (phi approximately phi(c)) than with conventional RF spoiling (phi >> phi(c)). As an example, the uptake of contrast agents in tissues is investigated. For typical parameters a considerably improved contrast enhancement can be obtained, both theoretically and experimentally. PMID- 16342264 TI - Mutations in alpha-helical solvent-exposed sites of eglin c have long-range effects: evidence from molecular dynamics simulations. AB - Eglin c is a small protease inhibitor whose structural and thermodynamic properties have been well studied. Previous thermodynamic measurements on mutants at solvent-accessible positions in the protein's helix have shown the unexpected result that the data could be best fit by the inclusion of residue- and position specific parameters to the model. To explore the origins of this surprising result, long molecular dynamics simulations in explicit solvent have been performed. These simulations indicate specific long-range interactions between the solvent-exposed residues in the eglin c alpha-helix and binding loop, an unexpected observation for such a small protein. The residues involved in the interaction are on opposite sides of the protein, about 25 A apart. Simulations of alanine substitutions at the solvent-exposed helix positions, arginine 22, glutamic acid 23, threonine 26, and leucine 27, show both small and large perturbations of eglin c dynamics. Two mutations exhibit large impacts on the long-range helix-loop interactions. Previous stability measurements (Yi et al., Biochemistry 2003;42:7594-7603) had indicated that an alanine substitution at position 27 was less stabilizing than at other solvent-exposed positions in the helix. The L27A mutation effects observed in these simulations suggest that the position-dependent loss of stability measured in wet bench experiments is derived from changes in dynamics that involve long-range interactions; thus, these simulations support the hypothesis that solvent-exposed positions in helices are not always equivalent. PMID- 16342265 TI - Pressure makes diamonds. PMID- 16342266 TI - Suggesting reviewers. PMID- 16342267 TI - Orthogonal tensor invariants and the analysis of diffusion tensor magnetic resonance images. AB - This paper outlines the mathematical development and application of two analytically orthogonal tensor invariants sets. Diffusion tensors can be mathematically decomposed into shape and orientation information, determined by the eigenvalues and eigenvectors, respectively. The developments herein orthogonally decompose the tensor shape using a set of three orthogonal invariants that characterize the magnitude of isotropy, the magnitude of anisotropy, and the mode of anisotropy. The mode of anisotropy is useful for resolving whether a region of anisotropy is linear anisotropic, orthotropic, or planar anisotropic. Both tensor trace and fractional anisotropy are members of an orthogonal invariant set, but they do not belong to the same set. It is proven that tensor trace and fractional anisotropy are not mutually orthogonal measures of the diffusive process. The results are applied to the analysis and visualization of diffusion tensor magnetic resonance images of the brain in a healthy volunteer. The theoretical developments provide a method for generating scalar maps of the diffusion tensor data, including novel fractional anisotropy maps that are color encoded for the mode of anisotropy and directionally encoded colormaps of only linearly anisotropic structures, rather than of high fractional anisotropy structures. PMID- 16342268 TI - Crystal structure of thioredoxin peroxidase from aerobic hyperthermophilic archaeon Aeropyrum pernix K1. PMID- 16342269 TI - Carotid stenting followed by coronary bypass: is drive-by coronary revascularization next? PMID- 16342270 TI - Staged carotid artery stenting and coronary artery bypass graft surgery: initial results from a single center. AB - The objective of this study was to assess the clinical course of patients undergoing planned percutaneous carotid stenting followed by staged coronary artery bypass grafting (CABG). Coexisting carotid and coronary atherosclerotic disease is relatively common. A combined or staged surgical approach has a composite stroke, myocardial infarction, or death rate of > 10%. We performed a retrospective search of our single-institution database to identify all patients scheduled to undergo staged carotid stenting followed by CABG. Twenty-three such patients (17 males, 6 females) were identified, with 3/23 (13%) requiring bilateral carotid stenting. Most carotid lesions were asymptomatic (18/26; 69.2%) and severe (mean stenosis, 82.9% 6+/- 8.6%). Stents were successfully placed in 26/26 carotid arteries (100%). One stent procedure (1/26; 3.8%) resulted in a minor stroke, but full recovery occurred within 1 week. There were no other peri stenting complications. Three patents (3/23; 13%), none of whom suffered an adverse event at carotid stenting, elected not to undergo CABG. The mean interval from last carotid stent to CABG was 69.6 6 +/- 39.6 days (range, 8-157 days). Antiplatelet therapy was ceased > 3 days prior to CABG in 10/20 patients (50%), but continued until surgery in the remainder. There were no peri-CABG bleeding or neurological complications, but one myocardial infarction occurred (1/20; 5%). Therefore, of the 20 patients who underwent planned carotid stenting followed by CABG, our overall rate of death, stroke, or myocardial infarction was 10%. However, our rate of death, persistent stroke or myocardial infarction was 5%. Planned carotid stenting followed by staged CABG is a viable method of treatment for patients with coexistent carotid and coronary atherosclerosis. PMID- 16342271 TI - Three dimensional CT angiography for patients with congenital heart disease: scanning protocol for pediatric patients. AB - The objective of our study was to determine the contrast attenuation level that yields high quality cardiac three-dimensional (3-D) images and to predict the contrast injection rate (IR), from body weight, to reach this attenuation level. Enhanced electron beam computerized tomography (EBCT) with 3-D reconstruction is useful in delineating cardiac anatomy in complex congenital heart disease (CHD). The current experience of using electron beam angiography (EBA) in pediatric CHD is limited. Well-defined contrast injection protocols, specifically the contrast IR, have not been standardized when compared to those for adults. Establishing the contrast IR is essential in obtaining high quality 3-D images. We retrospectively analyzed the studies of 115 pediatric patients with CHD. EBA images were divided into group 1 with good quality 3-D images and group 2 with poor quality. The mean of measured enhancement level, expressed in Hounsfield units (HU), and contrast IR were analyzed in both groups. Spearman correlation was used to examine the relationship between weight and IR. The IR was predicted from weight using simple linear regression analysis. The mean level of enhancement was 344 +/- 91 and 174 +/- 31 HU for group 1 and group 2, respectively. Group 1 consisted of 103 patients (90%) and the IR strongly correlated with weight (rho = 0.861, P < 0.01). The IR was estimated from the linear regression equation IR = 0.59 + 0.056 x weight. Necessary contrast enhancement level for quality 3-D reconstruction should be greater than 250 HU, and the IR can be estimated from patient's weight. PMID- 16342272 TI - Computational reconstruction of the human skeletal muscle secretome. AB - In multicellular organisms, secreted proteins play pivotal regulatory roles in intercellular communication. Proteins secreted by skeletal muscle can act locally on muscle cells through autocrine/paracrine loops and on surrounding tissues such as muscle blood vessels, or they can be released into the blood stream, thus producing systemic effects. By a computational approach, we have screened 6255 products of genes expressed in normal human skeletal muscle. Putatively secreted proteins were identified by sequential steps of sieving, through prediction of signal peptide, recognition of transmembrane regions, and analysis of protein annotation. The resulting putative skeletal muscle secretome consists of 319 proteins, including 78 still uncharacterized proteins. This is the first human skeletal muscle secretome produced by computational analysis. Knowledge of proteins secreted by skeletal muscle could stimulate development of novel treatments for different diseases, including muscle atrophy and dystrophy. In addition, better knowledge of the secretion process in skeletal muscle can be useful for future gene therapy approaches. PMID- 16342273 TI - Using fMRI to dissociate sensory encoding from cognitive evaluation of heat pain intensity. AB - Neuroimaging studies of painful stimuli in humans have identified a network of brain regions that is more extensive than identified previously in electrophysiological and anatomical studies of nociceptive pathways. This extensive network has been described as a pain matrix of brain regions that mediate the many interrelated aspects of conscious processing of nociceptive input such as perception, evaluation, affective response, and emotional memory. We used functional magnetic resonance imaging in healthy human subjects to distinguish brain regions required for pain sensory encoding from those required for cognitive evaluation of pain intensity. The results suggest that conscious cognitive evaluation of pain intensity in the absence of any sensory stimulation activates a network that includes bilateral anterior insular cortex/frontal operculum, dorsal lateral prefrontal cortex, bilateral medial prefrontal cortex/anterior cingulate cortex, right superior parietal cortex, inferior parietal lobule, orbital prefrontal cortex, and left occipital cortex. Increased activity common to both encoding and evaluation was observed in bilateral anterior insula/frontal operculum and medial prefrontal cortex/anterior cingulate cortex. We hypothesize that these two regions play a crucial role in bridging the encoding of pain sensation and the cognitive processing of sensory input. PMID- 16342274 TI - Evaluating the quality of NMR structures by local density of protons. AB - Evaluating the quality of experimentally determined protein structural models is an essential step toward identifying potential errors and guiding further structural refinement. Herein, we report the use of proton local density as a sensitive measure to assess the quality of nuclear magnetic resonance (NMR) structures. Using 256 high-resolution crystal structures with protons added and optimized, we show that the local density of different proton types display distinct distributions. These distributions can be characterized by statistical moments and are used to establish local density Z-scores for evaluating both global and local packing for individual protons. Analysis of 546 crystal structures at various resolutions shows that the local density Z-scores increase as the structural resolution decreases and correlate well with the ClashScore (Word et al. J Mol Biol 1999;285(4):1711-1733) generated by all atom contact analysis. Local density Z-scores for NMR structures exhibit a significantly wider range of values than for X-ray structures and demonstrate a combination of potentially problematic inflation and compression. Water-refined NMR structures show improved packing quality. Our analysis of a high-quality structural ensemble of ubiquitin refined against order parameters shows proton density distributions that correlate nearly perfectly with our standards derived from crystal structures, further validating our approach. We present an automated analysis and visualization tool for proton packing to evaluate the quality of NMR structures. PMID- 16342275 TI - Highly constrained backprojection for time-resolved MRI. AB - Recent work in k-t BLAST and undersampled projection angiography has emphasized the value of using training data sets obtained during the acquisition of a series of images. These techniques have used iterative algorithms guided by the training set information to reconstruct time frames sampled at well below the Nyquist limit. We present here a simple non-iterative unfiltered backprojection algorithm that incorporates the idea of a composite image consisting of portions or all of the acquired data to constrain the backprojection process. This significantly reduces streak artifacts and increases the overall SNR, permitting decreased numbers of projections to be used when acquiring each image in the image time series. For undersampled 2D projection imaging applications, such as cine phase contrast (PC) angiography, our results suggest that the angular undersampling factor, relative to Nyquist requirements, can be increased from the present factor of 4 to about 100 while increasing SNR per individual time frame. Results are presented for a contrast-enhanced PR HYPR TRICKS acquisition in a volunteer using an angular undersampling factor of 75 and a TRICKS temporal undersampling factor of 3 for an overall undersampling factor of 225. PMID- 16342276 TI - Folding and amyloid-fibril formation for a series of human stefins' chimeras: any correlation? AB - To study the influence of whole secondary structure elements to the process of folding and amyloid-fibril formation, chimeras of stefins have been prepared. GdnHCl denaturation curves and folding rates (chevron plots) have been analyzed based on a two-state mechanism. The order of stability is: stefin A > aAbbbb > bAbbbb > stefin B = aBaaaa > bBaaaa, where the make up of chimeric proteins is designated by small letters representing the source of individual strands (a for stefin A, b for stefin B) and a capital letter representing the source of the helix (A for stefin A and B for stefin B). Only the fast folding reactions were included in the analysis and it has been found that stefin B folds the fastest (657 s(-1)). Similarly, fast folders are the chimeric proteins aBaaaa and bBaaaa, both of which contain the alpha-helix of stefin B. Unfolding rates correlate very well with protein stability, with the slowest rate for the most stable protein, stefin A. Amyloid-fibril growth was measured for each protein by monitoring thioflavin T fluorescence and was visualized using electron microscopy. The propensity to form amyloid-fibrils is in the order: stefin B > bAbbbb > aAbbbb > bBaaaa > aBaaaa > stefin A. This order does not correlate with stability, or with the folding or unfolding rates. Instead, the propensity to fibrillize is related to selected parts of structure, such as the beta-sheet of stefin B, and can be predicted reasonably well by calculating the beta-strand propensity of the denatured states. PMID- 16342277 TI - Oxygen enhances lethal effect of high-intensity, ultrashort electrical pulses. AB - The study explored the effect of ambient oxygen on mammalian cell survival after exposure to 10 ns duration, high voltage electrical pulses (nsEP, 80-90 or 120 130 kV/cm; 200-400 pulses per exposure). Cell samples were equilibrated with pure nitrogen, atmospheric air, or pure oxygen prior to the nsEP treatment and were returned to the incubator (air + 5% CO2) shortly after the exposure. The experiments established that survival of hypoxic Jurkat and U937 cells exceeded that of air-equilibrated controls about twofold (P < .01). Conversely, saturation of the medium with oxygen prior to exposure decreased Jurkat cell survival about 1.5 times, P < .01. Attenuation of the cytotoxic effect under hypoxic conditions resembled a well-known effect of oxygen on cell killing by sparsely ionizing radiations and may be indicative of the similarity of underlying cell damage mechanisms. PMID- 16342278 TI - Trichloroethylene and dichloroethylene: a critical review of teratogenicity. AB - Trichloroethylene (TCE) and dichloroethylene (DCE) are high-volume industrial chemicals frequently found as contaminants in public drinking water supplies. The developmental toxicity of both chemicals has been evaluated in laboratory and epidemiologic studies. It has been suggested that TCE and DCE are specific cardiac teratogens and that drinking water contaminated with them increases the risk of congenital heart defects in exposed human populations. In contrast, other laboratory and epidemiologic studies do not find an increase in developmental effects, either in general or specifically affecting the heart. This laboratory and epidemiologic base was reviewed to evaluate the strengths and weaknesses of the conflicting published reports. We conclude that the weight of experimental and epidemiologic evidence does not support the hypothesis that TCE or DCE is a selective developmental toxicant in general or a cardiac teratogen specifically. PMID- 16342279 TI - Genotypic interaction between DRD4 and DAT1 loci is a high risk factor for attention-deficit/hyperactivity disorder in Chilean families. AB - Attention-deficit/hyperactivity disorder, ADHD [MIM 126452], is a common, highly heritable neurobiological disorder of childhood onset, characterized by hyperactivity, impulsiveness, and/or inattentiveness. As part of an ongoing study of ADHD, we carried out a family-based discordant sib-pair analysis to detect possible associations between dopamine receptor D4 (DRD4) and dopamine transporter 1 (DAT1) polymorphisms and ADHD in Chilean families. Both loci individually classified as homozygotes or heterozygotes for the DRD4 7-repeat and DAT1 10-repeat alleles, did not exhibit genotype frequency differences between affected children and their healthy siblings (Fisher's exact test P > 0.25 in both cases). However, the simultaneous presence of both DRD4 7-repeat heterozygosity and DAT1 10 allele homozygosity were significantly higher (34.6%) in cases (26), compared with their unaffected siblings (25) (4%; Fisher's exact test P = 0.0096; odds-ratio, OR = 12.71). Increased density of dopamine transporter in ADHD brains, along with abundance of 7-repeat D4 receptors in prefrontal cortex, which is impaired in ADHD patients, make the observed gene gene interaction worthy of further incisive studies. PMID- 16342280 TI - Human QKI, a new candidate gene for schizophrenia involved in myelination. AB - We have previously shown that chromosome 6q25-6q27 includes a susceptibility locus for schizophrenia in a large pedigree from northern Sweden. In this study, we fine-mapped a 10.7 Mb region, included in this locus, using 42 microsatellites or SNP markers. We found a 0.5 Mb haplotype, likely to be inherited identical by decent, within the large family that is shared among the majority of the patients (69%). A gamete competition test of this haplotype in 176 unrelated nuclear families from the same geographical area as the large family showed association to schizophrenia (empirical P-value 0.041). The only gene located in the region, the quaking homolog, KH domain RNA binding (mouse) (QKI), was investigated in human brain autopsies from 55 cases and 55 controls using a high-resolution mRNA expression analysis. Relative mRNA expression levels of two QKI splice variants were clearly downregulated in schizophrenic patients (P-value 0.0004 and 0.03, respectively). The function of QKI has not been studied in humans, but the mouse homolog is involved in neural development and myelination. In conclusion, we present evidence from three unrelated sample-sets that propose the involvement of the QKI gene in schizophrenia. The two family based studies suggest that there may be functional variants of the QKI gene that increase the susceptibility of schizophrenia in northern Sweden, whereas the case-control study suggest that splicing of the gene may be disturbed in schizophrenic patients from other geographical origins. Taken together, we propose QKI as a possible target for functional studies related to the role of myelination in schizophrenia. PMID- 16342281 TI - Genetic liability to schizophrenia or bipolar disorder and its relationship to brain structure. AB - Bipolar disorder and schizophrenia are highly heritable conditions that are associated with structural brain abnormalities. Although brain abnormalities are found in the well relatives of people with schizophrenia, the extent to which genetic liability relates to brain structure in either disorder is still unclear. This study sought to ascertain the effects of genetic liability to schizophrenia and bipolar disorder on white and grey matter volume in patients with these diagnoses and their well relatives. Seventy-one patients and 72 unaffected relatives were recruited for the study. Patients included those with schizophrenia from families affected by schizophrenia alone, those with bipolar disorder from families affected by bipolar disorder alone and those with bipolar disorder from families affected by both bipolar disorder and schizophrenia. Samples of unaffected relatives of each patient group were also recruited. Subjects underwent an MRI scan of the brain, which was analysed using optimised voxel-based morphometry (VBM). Grey and white matter volume was then related to a continuous measure of genetic liability based on a threshold-liability model. Genetic liability to schizophrenia was associated with decreased grey matter volume in dorso- (DLPFC) and ventrolateral prefrontal (VLPFC) cortices. The relationship remained after diagnostic status had been taken into account. Complementary white matter changes were also demonstrated. No relationship was demonstrated between a genetic liability to bipolar disorder and either white or grey matter volume. Genes that raise the likelihood of developing schizophrenia may exert their effects by diminishing grey matter volume in the DLPFC and VLPFC and their associated white matter connections. Genes for bipolar illness might have subtle effects on brain structure, which may need particularly large samples to detect. PMID- 16342282 TI - Association study of a functional promoter polymorphism in the XBP1 gene and schizophrenia. AB - A functional promoter polymorphism (-116C/G) of the X-box binding protein 1 gene (XBP1) gene was reported to be associated with schizophrenia in Asian subjects. In a replication attempt, three European case-control samples comprising 2,182 German, Polish, and Swedish subjects, were genotyped for the XBP1 -116C/G polymorphism. Allele and genotype frequencies were compared between schizophrenic patients and control subjects. There were no significant case-control differences in any of the three samples, although in a meta-analysis with previous results comprising 3,612 subjects there was a borderline association between the -116G containing genotypes and schizophrenia. We conclude that the functional XBP1 gene polymorphism is not of major importance to schizophrenia in the European populations investigated. It cannot be excluded, however, that the XBP1 polymorphism is involved in schizophrenia in other populations or adds minor susceptibility to the disorder. PMID- 16342283 TI - Maternal age at the birth of the first child as an epistatic factor in polygenic disorders. AB - The identification of the genes for complex, polygenic disorders has proven difficult. This is due to the small effect size of each gene and genetic heterogeneity. An additional important factor could be the presence of unidentified epistatic factors. In the broad definition of epistasis, the effect of one unit is not predicable unless the value of another unit is known and one of the units may not be a gene. We have previously identified maternal age as an epistatic factor for the effect of the LEP gene on the age of onset of menarche. We report here the effect of maternal age and the age of the mother at the birth of her first child (maternal age 1st) as epistatic factors for the interaction of the dopamine D1 gene (DRD1) with obsessive-compulsive behaviors and with stuttering. The epistatic effects of maternal age 1st were stronger than maternal age. This type of epistatic factor may be generalizable to many other gene-trait interactions. PMID- 16342284 TI - Bilateral mirror writing movements (mirror dystonia) in a patient with writer's cramp: functional correlates. AB - A recent prospective analysis on writer's cramp showed that up to 44.6% of patients in a series of 65 presented mirror dystonia, defined as involuntary movements of the resting hand, abnormal posture, tremor, and jerks occurring while writing with the opposite hand. A clinical case is presented, with videotape evidence of right-handed writer's cramp, with mirror movements elicited while writing using either hand. Functional magnetic resonance imaging studies are compared both to those of a normal patient and to those from a patient with writer's cramp but lacking mirror dystonia. Widespread bilateral activation of cortical motor areas contralateral to the mirror movements in patients with writer's cramp and mirror movements suggests, that bilateral activation of the primary motor cortex may account for the appearance of these mirror movements. Further studies need to be conducted to determine whether mirror movements in dystonic patients appear as a result of loss of intra- and/or interhemispheric cortical inhibition or are simply a consequence of the sustained effort these patients need to exert while writing using a dystonic hand. PMID- 16342285 TI - Risk management with suicidal patients. AB - The patient who is at-risk for suicide is complex and is difficult to evaluate and treat effectively. Should suicidal behavior occur, the clinician faces the potential wrath of bereaved survivors and their externalized blame exercised through a malpractice suit. The clinician's duty of care to a patient is to act affirmatively to protect a patient from violent acts against self. A finding of malpractice is established if the court finds that this duty was breached, through an act of omission or commission relative to the standard of care, and that this breach was proximately related to the patient's suicidal behavior. This article discusses the standard of care and factors that determine liability in a suicide death of a patient. An extensive list of recommendations for competent caregiving for the at-risk patient and risk management guidelines are then presented. PMID- 16342286 TI - Suicidality in clinical practice: anxieties and answers. AB - This issue of the Journal of Clinical Psychology: In Session is devoted to suicidality in clinical practice, with coverage of a broad range of topics. After summarizing some of the problems and recent advances in clinical suicidology, I provide a brief overview of the articles. Included are the more broadly defined issues of risk management and psychotherapy with suicidal individuals, along with more specific topics such as risk assessment, no-suicide contracts, and the risk of suicide with the use of selective serotonin reuptake inhibitors (SSRIs) by children and adolescents. Most of the articles feature case vignettes and provide practitioners with clinical recommendations, and all of the articles have clear and important clinical implications. PMID- 16342287 TI - Mindfulness-based cognitive therapy for prevention of recurrence of suicidal behavior. AB - Once suicidal thoughts have emerged as a feature of depression they are likely to be reactivated as part of a suicidal mode of mind whenever sad mood reappears. This article reviews the methods and the usefulness of mindfulness-based cognitive therapy (MBCT) as a treatment for the prevention of the reactivation of the suicidal mode. MBCT integrates mindfulness meditation practices and cognitive therapy techniques. It teaches participants to develop moment-by-moment awareness, approaching ongoing experience with an attitude of nonjudgment and acceptance. Participants are increasingly able to see their thoughts as mental events rather than facts (metacognitive awareness). A case example illustrates how mindfulness skills develop with MBCT and how they relate to the cognitive processes that fuel suicidal crises. An ongoing controlled trial will provide further evidence, but pilot work suggests that MBCT is a promising intervention for those who have experienced suicidal ideation in the past. PMID- 16342288 TI - Advances in the assessment of suicide risk. AB - This article reviews and integrates empirically grounded advances in the assessment of suicidality. The practices discussed are consistent with existing standards of care, practice guidelines, and applicable research. The authors differentiate between risk assessment and prediction and then emphasize the important role of time in risk assessment. We present and illustrate a continuum of suicidality for risk assessment and offer practical recommendations for clinical decision making and treatment. PMID- 16342289 TI - Clinical applications of the interpersonal-psychological theory of attempted and completed suicide. AB - This article reviews the interpersonal-psychological theory of attempted and completed suicide and describes its applications in suicide risk assessment, crisis intervention, and skills-based psychotherapies. Three components are necessary, but not sufficient, for an individual to die by suicide: (1) the acquired capability to enact lethal self-injury, (2) a sense that one is a burden on others, and (3) the sense that one does not belong to a valued social group. We suggest that therapeutic interventions should focus on ascertaining the presence of these components and work to amend the cognitive distortions, negative interpersonal response styles, and ineffective coping behaviors that serve to maintain suicidal urges. PMID- 16342290 TI - Psychoanalytic perspectives on the treatment of an acute suicidal crisis. AB - The psychoanalytic understanding of suicide has greatly expanded in the past 30 years, and the older therapeutic approach has been modified. Although the turning of murder on the self inferred by Freud 90 years ago remains a cornerstone, current practice concentrates on the identification of a precipitating event, the escalation of intolerable painful affect that the patient cannot moderate, the self-perception of helplessness, the evolution of hopelessness as a secondary affect, increasing fantasies of suicide as a means of escape, and ego regression (self-deconstruction), leading progressively to suicide. Recent clinical research, advances in the study of cognition, and neurobiological studies have contributed to the reorientation and enrichment of the psychoanalytic perspective on suicide. The treatment of a university student who had fallen into a suicide crisis is discussed from the psychoanalytic point of view, and evidence for the efficacy of this treatment is reviewed. PMID- 16342291 TI - Do SSRIs cause suicide in children? The evidence is underwhelming. AB - After regulatory agencies in the United Kingdom and United States recommended severe restrictions on antidepressant use in children, many lessons were learned, although one was not that these drugs cause suicide. We learned that pharmaceutical companies selectively released data that reflected positively on their products and that combining suppressed and published data suggested that most of these medications had questionable efficacy. We also learned that the studies lacked uniformity both in which age groups constituted children and which behavior was considered suicidal. Several recent, large nonindustry studies indicated that rates of suicide and suicidal behavior were actually reduced in children who used antidepressants, despite piteous anecdotal tales in the popular press purporting that selective serotonin reuptake inhibitors (SSRIs) caused children to kill themselves. Patients in pharmaceutical trials probably do not represent typical patients in routine clinical practice. Emerging implications are that suicidal behavior-if it does occur-is most likely soon after starting antidepressant use and that prescribers must be both vigilant in educating patients and families about warning signs and available to manage worrisome behavior. PMID- 16342292 TI - Psychosocial treatments of suicidal behaviors: a practice-friendly review. AB - Worldwide, almost a million people die by suicide each year. Intentional, nonfatal, self-inflicted injury, including both suicide attempts and acts without suicide intent, also has very high prevalence. This article provides a practice friendly review of controlled studies of psychosocial treatments aiming to prevent suicide, attempted suicide, and nonsuicidal self-inflicted injuries. Despite relatively small sample sizes for a low-base-rate outcome such as self inflicted injury, several psychotherapies have been found effective, including cognitive therapy, dialectical behavior therapy, problem-solving therapy, and interpersonal psychotherapy, as well as outreach interventions, such as sending caring letters. The clinical implications of the review are discussed with the goal of translating the science to service-particularly the importance of outreach and treatment of non-compliance, the assessment and management of suicide risk, and competency in effective psychotherapies. These are critical steps for clinical psychology and psychotherapists to take in their role in suicide prevention. PMID- 16342293 TI - The case against no-suicide contracts: the commitment to treatment statement as a practice alternative. AB - This article reviews the literature on the use of "no-suicide contracts" in clinical practice, including conceptual discussions, patient and clinician surveys, and a few empirical studies on clinical utility. Our primary conclusion is that no-suicide contracts suffer from a broad range of conceptual, practical, and empirical problems. Most significantly, they have no empirical support for their effectiveness in the clinical environment. The authors provide and illustrate the commitment to treatment statement as a practice alternative to the no-suicide contract. PMID- 16342294 TI - Associations of demographic and lifestyle characteristics with prostate-specific antigen (PSA) concentration and rate of PSA increase. AB - BACKGROUND: The objective of this study was to examine whether demographic and lifestyle characteristics are associated with prostate-specific antigen (PSA) levels and the rate of PSA increase (PSA velocity). METHODS: Data for this study came from 3341 participants in the placebo arm of the Prostate Cancer Prevention Trial who, based on biopsies at the end of the study, were free of prostate carcinoma. Linear regression was used to assess associations of age, race, smoking, body mass index (BMI), physical activity, diet, and supplement use with PSA concentration during the second year of the trial, and linear mixed models were used to assess associations of these factors with PSA velocity (the percentage increase in PSA per year) during 6 years of the trial. RESULTS: Between the group of men ages 50-59 years and the group of men age 70 years and older, mean PSA increased by 0.22 ng/mL, and PSA velocity decreased by 1.2 percentage points (both P < 0.001). The PSA level among men who had a BMI > or = 35 kg/cm(2) was 0.20 ng/mL lower than the PSA level among men who had a BMI < 25 kd/cm(2) (P < 0.001), but BMI was not associated with PSA velocity. PSA velocity was 1.2 percentage points higher in African-American men compared with white men (P = 0.043). Low energy intake and the use of high-dose calcium supplements were associated with significantly lower PSA velocity (both P = 0.05). Weight gain also was associated with lower PSA velocity. CONCLUSIONS: Differences in PSA concentration associated with demographic and lifestyle characteristics were small and were not likely to bias the interpretation of a single PSA test. Age, race, energy intake, calcium supplement use, and weight change were associated with substantial differences in PSA velocity, and the clinical interpretation of PSA velocity may be biased by these factors. PMID- 16342295 TI - Predictors of enrollment in lung cancer clinical trials. AB - BACKGROUND: Clinical trials may offer patients innovative therapeutic options with potentially better outcomes, which are particularly relevant for patients afflicted with lung carcinoma, because current therapies provide only modest survival benefits. Only approximately 5% of patients with newly diagnosed cancer participate in clinical trials nationwide, and African-American (AA) patients are particularly under-represented. METHODS: To determine predictors of clinical trials enrollment, the authors reviewed the medical records of 427 patients with lung carcinoma (175 AA patients and 252 non-AA patients) who were eligible for clinical trials between 1994 and 1998 at the Karmanos Cancer Institute in Detroit, Michigan. Logistic regression analysis was used to assess the association of patient demographic characteristics and clinical trial enrollment. RESULTS: Ninety-one patients (21%) were enrolled onto a lung cancer clinical trial during the period of the current study. Enrollment was associated significantly with race (P < 0.001), gender (P = 0.048), age (P = 0.005), and insurance type (P = 0.024). After multivariable adjustment, only race and gender remained significant predictors of enrollment. AA patients were less likely to enroll than non-AA patients (odds ratio [OR], 0.485; 95% confidence interval [95% CI], 0.243-0.966), and men were more likely than women to enroll (OR, 1.812; 95% CI, 1.033-3.178). CONCLUSIONS: The current results suggest disparities by race and gender in the enrollment of patients onto lung cancer clinical trials and support the need to improve educational and outreach endeavors that would make clinical trials available to a wider range of eligible patients. PMID- 16342296 TI - Prognostic impact of cytogenetic aberrations in patients with multiple myeloma or monoclonal gammopathy of unknown significance. AB - Chromosomal aberrations are the most important prognostic factors in haematological malignancies. Detection of certain genetic changes leads to risk adapted strategies in leukaemia therapy. In multiple myeloma the importance of genetic alterations and their prognostic impact is of growing interest. Several therapeutic approaches seem to be uneffective for patients harbouring certain chromosomal abnormalities. Although the yield of metaphases due to a low proliferation rate is considerably lower in plasma cell dyscrasias, a number of chromosomal changes with prognostic implications have been identified in the past years, particularly due to the introduction of new techniques. This article gives a short survey of the most important genetic alterations and their prognostic influence on the outcome of patients with plasma cell malignancies known to date. PMID- 16342297 TI - Ionizing radiation and leukaemia: more questions than answers. AB - The mechanisms underlying the unequivocal association between ionizing radiation and the development of leukaemia remain unknown. Recent progress in defining sub cellular events has contributed to our understanding of the production of genetic lesions in irradiated cells but the importance of tissue effects in response to radiation damage has attracted much less attention. Thus, genetic lesions induced by radiation are considered to result from the deposition of energy in the cell nucleus and the initiating lesion for radiation-induced transformation has been similarly attributed to direct DNA damage. Recently, however, there have been many reports of radiation effects, characteristically associated with the consequences of energy deposition in the cell nucleus, arising in non-irradiated cells as a consequence of communication with irradiated cells. These, so-called, non-targeted radiation effects pose major challenges to current views of the mechanisms of radiation-induced DNA damage and the mechanisms underlying radiogenic malignancies. Considered together with data obtained from laboratory model systems, a rather complex picture of radiation leukaemogenesis is emerging in which, additional to any damage induced directly in target stem cells, the haemopoietic microenvironment can be a source of damaging signals and cellular interactions make important genotype-dependent contributions to determining overall outcome after radiation exposures. PMID- 16342298 TI - Evidence of abortive plasma cell differentiation in Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma. AB - Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (cHL) show genotypic features of germinal centre-derived B-cells in most cases. Nevertheless, these cells typically lack expression of B-cell antigens. Previous studies have suggested that plasma cell differentiation may occur in HRS cells and that this may account for the down-regulation of B-cell antigens. However, these results are controversial. We have addressed this question using immunohistochemistry and a panel of antibodies directed against antigens which are differentially expressed during terminal B-cell differentiation. Pax-5, a transcription factor required for B-lineage commitment, and IRF4/Mum1, which is physiologically expressed in germinal centre cells and plasma cells, were consistently detectable in HRS cells. Bcl-6, a transcription factor expressed in germinal centre B-cells, was present in HRS cells of approximately 25% of cHL cases. Expression of the B-lymphocyte-induced maturation protein-1 (Blimp-1), a key regulator of plasma cell differentiation, was observed in HRS cells of 23% of cHL cases. In these cases, Blimp-1 expression was restricted to a small proportion of HRS cells. HRS cells were consistently negative for the plasma cell marker CD138. These results suggest that plasma cell differentiation may be initiated in a small subset of HRS cells but remains abortive. Thus, terminal differentiation is unlikely to explain the lack of B-cell antigen expression in HRS cells. PMID- 16342299 TI - Pharmacoepidemiology of antihypertensive drugs in primary care setting of Bahrain between 1998 and 2000. AB - PURPOSE: To compare pattern of antihypertensive drug utilization in 1998 with 2000 following the: (a) publication of 1999 World Health Organization/International Society of Hypertension (WHO/ISH) guidelines for drug management of hypertension; and (b) introducing new antihypertensives to the essential drug list, in primary care, Bahrain. METHODS: Retrospective prescription-based survey carried out in seven out of 20 primary care health centers in Bahrain. A total of 9272 patients comprising 6543 with uncomplicated hypertension and 2729 with diabetic hypertension were studied. RESULTS: Between 1998 and 2000, the prescription rate of beta-blockers and methyldopa significantly declined (p < 0.0001) while the rate of angiotensin converting enzyme (ACE) inhibitors and calcium channel blockers (CCBs) increased (p < 0.0001) in uncomplicated hypertension. However, in diabetic hypertension there was a non-significant decline in utilization of beta-blockers, CCBs, methyldopa, and diuretics whereas a significant increase in prescribing of ACE inhibitors (p < 0.0001). Inclusion of Perindopril into the essential drug list resulted in an overall increase in utilization of ACE inhibitors: use of captopril and enalapril declined while lisinopril and perindopril increased. Substitution of immediate release (IR) indapamide by sustained-release (SR) formulation did not change the overall utilization of diuretics; however, intra-class changes were evident with a significant decline in use of thiazide diuretics and concomitant increase in use of SR indapamide by 2000. CONCLUSIONS: The antihypertensive prescribing pattern is influenced by WHO/ISH guidelines as well as by introduction of new antihypertensives to primary care essential drug list in primary care. The choice of a drug is also influenced by presence of co-morbidity with diabetes mellitus. PMID- 16342300 TI - Photochemically enhanced transduction of polymer-complexed adenovirus targeted to the epidermal growth factor receptor. AB - BACKGROUND: The development of methods for specific delivery of genes into target tissues is an important issue for the further progress of gene therapy. Biological and physical targeting techniques may be combined to redirect gene therapy vectors to specific cells and enhance gene transfer. METHODS: The polymer poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) was conjugated with avidin or poly(ethylene glycol) (PEG) and complexed with adenovirus serotype 5 (Ad5). Targeting of polymer-coated Ad5 to the epidermal growth factor receptor (EGFR) was accomplished by the binding of biotin-EGF to pDMAEMA-avidin. A photochemical treatment procedure using photosensitizer and light was applied to increase transduction with EGFR-targeted viral complexes. RESULTS: pDMAEMA-avidin efficiently enhanced transduction through unspecific viral uptake into cells, while pDMAEMA-PEG provided charge shielding of the complexes and increased the specificity to EGFR when biotin-EGF ligands were used. Transduction of PEG containing, EGFR-targeted viral complexes was inhibited by 66% in coxsackie and adenovirus receptor (CAR)-deficient RD cells and by 47% in CAR-expressing DU 145 cells in receptor antibody experiments. The photochemical treatment had a substantial effect on transduction, enhancing the percentage of reporter gene positive cells from 20% to 75% of the total viable RD cell population and from 10% to 70% in DU 145 cells. CONCLUSION: Photochemical treatment of cells infected with targeted viral vectors exhibiting a neutral surface charge is a potent method for enhancing transgene expression. PMID- 16342302 TI - Effect of intervention through a pharmaceutical care program on patient adherence with prescribed once-daily atorvastatin. AB - AIMS: Correct execution of prescribed dosing regimen(s) is essential for patients to benefit from lipid lowering treatments. The objective of this study was to estimate the effect of a pharmaceutical care program on the adherence of once daily atorvastatin treatment in patients with elevated cholesterol levels. METHODS: In both linguistic regions of Belgium, two districts were randomized between usual care and a supportive intervention program. Eligible patients included hyperlipemic subjects taking atorvastatin for at least 3 months. 'Adherence' was defined as the proportion of days during which the electronic device record showed that the patient had taken the daily dose. 'Persistence' quantifies how long the treatment is executed. RESULTS: A total of 392 patients from 35 pharmacies were included in the intent-to-treat (ITT) analysis of the data (194 patients received intervention and 198 in the control group). The intervention resulted in a 6.5% increase in post-baseline adherence (p < 0.001) mainly driven by a 13% increase in persistence at 1 year (p = 0.002). CONCLUSION: A supportive pharmaceutical care program consisting of review by patient and pharmacist of each patient's electronically compiled dosing history, plus educational reminders, improves patient adherence to the once-daily atorvastatin dosing regimen, mainly by extending persistence. PMID- 16342303 TI - Directed evolution of Bacillus subtilis lipase A by use of enantiomeric phosphonate inhibitors: crystal structures and phage display selection. AB - Phage display can be used as a protein-engineering tool for the selection of proteins with desirable binding properties from a library of mutants. Here we describe the application of this method for the directed evolution of Bacillus subtilis lipase A, an enzyme that has important properties for the preparation of the pharmaceutically relevant chiral compound 1,2-O-isopropylidene-sn-glycerol (IPG). PCR mutagenesis with spiked oligonucleotides was employed for saturation mutagenesis of a stretch of amino acids near the active site. After expression of these mutants on bacteriophages, dual selection with (S)-(+)- and (R)-(-)-IPG stereoisomers covalently coupled to enantiomeric phosphonate suicide inhibitors (SIRAN Sc and Rc inhibitors, respectively) was used for the isolation of variants with inverted enantioselectivity. The mutants were further characterised by determination of their Michaelis-Menten parameters. The 3D structures of the Sc and Rc inhibitor-lipase complexes were determined and provided structural insight into the mechanism of enantioselectivity of the enzyme. In conclusion, we have used phage display as a fast and reproducible method for the selection of Bacillus lipase A mutant enzymes with inverted enantioselectivity. PMID- 16342304 TI - Optimization of xenon biosensors for detection of protein interactions. AB - Hyperpolarized 129Xe NMR spectroscopy can detect the presence of specific low concentration biomolecular analytes by means of a xenon biosensor that consists of a water-soluble, targeted cryptophane-A cage that encapsulates the xenon. In this work, we use the prototypical biotinylated xenon biosensor to determine the relationship between the molecular composition of the xenon biosensor and the characteristics of protein-bound resonances. The effects of diastereomer overlap, dipole-dipole coupling, chemical-shift anisotropy, xenon exchange, and biosensor conformational exchange on the protein-bound biosensor signal were assessed. It was found that an optimal protein-bound biosensor signal can be obtained by minimizing the number of biosensor diastereomers and using a flexible linker of appropriate length. Both the line width and sensitivity of chemical shift to protein binding of the xenon biosensor were found to be inversely proportional to linker length. PMID- 16342305 TI - Unsaturated fatty alcohol derivatives as a source of substituted allylzirconocene. PMID- 16342306 TI - "Perovskitization"-assisted electrochemical reduction of solid TiO2 in molten CaCl2. PMID- 16342307 TI - Co(II) chemistry of 2,6-bis(2-pyridylcarbonyl)pyridine: an icosanuclear Co cluster exhibiting superparamagnetic relaxation. PMID- 16342308 TI - Catalytic enantioselective cross-Mannich reaction of aldehydes. PMID- 16342309 TI - The Yandulov/Schrock cycle and the nitrogenase reaction: pathways of nitrogen fixation studied by density functional theory. PMID- 16342310 TI - Hydrophobically directed organic synthesis. PMID- 16342311 TI - Mechanistic studies on the conversion of arylamines into arylnitro compounds by aminopyrrolnitrin oxygenase: identification of intermediates and kinetic studies. PMID- 16342312 TI - Synthesis and conformational study of water-soluble, rigid, rodlike oligopiperidines. PMID- 16342313 TI - Gold-catalyzed cyclization of enynes. PMID- 16342314 TI - Transition-metal doped zinc oxide nanowires. PMID- 16342315 TI - Molecular design of neutral intramolecular complexes bearing two silicon atoms anchored by a carbonyl oxygen atom: N,N'-bis(silylmethyl)propylene ureas. AB - B3LYP and MP2 computations have been performed on a variety of Si,Si'-substituted N,N'-bis(silylmethyl)propylene ureas. According to electron-density atoms-in molecules (AIM) and electron localization function (ELF) quantum-topological analyses, a transition from the unstable non-chelate forms of these compounds to mono- and bis-chelate forms results in the successive interaction of one and two tetracoordinate silicon atoms with the carbonyl oxygen and the formation, respectively, of one and two covalent, polar Si...O bonds. This previously unknown X-Si<--O-->Si-X type of bonding in isomers possessing an anchor structure may be classified as a five-center, six-electron (5c-6e) bond. The factors that favor the existence of Si,Si'-substituted N,N'-bis(silylmethyl)propylene ureas exclusively in the form of stable, bridged complexes (the size of equatorial ligands and the electronegativity of axial substituents at the silicon atom, change in the donor capability of the carbonyl group, and effect of the polar solvent) are discussed. PMID- 16342316 TI - Synthesis and some properties of binuclear ruthenocenes bridged by oligoynes: formation of bis(cyclopentadienylidene)cumulene diruthenium complexes in the two electron oxidation. AB - The monoynes [Rc*C[triple bond]CRc*] and [Rc'C[triple bond]CRc'] were obtained in improved yields using [Mo(CO)6]/2-FC6H5OH as a catalyst in the alkyne metathesis of [Rc*C[triple bond]CMe] and [Rc'C[triple bond]CMe], respectively (Rc = ruthenocenyl, Rc* = 1'',2'',3'',4'',5''-pentamethylruthenocenyl, and Rc' = 2',3',4',5'-tetramethylruthenocenyl groups). The diynes [Rc*(C[triple bond]C)2Rc*] and [Rc'(C[triple bond]C)2Rc'] were synthesized by the oxidative coupling of the corresponding terminal ethynes in good yields. The triyne [Rc*(C[triple bond]C)3Rc*] and the tetrayne [Rc*(C[triple bond]C)4Rc*] were prepared by the hetero- and homocoupling of [Rc*C[triple bond]CC[triple bond]CH], which was obtained from the reaction of [Rc*C[triple bond]CCHO] with Li[N2CSiMe3], respectively. Although the oxidation waves did not always exhibit a clear two-electron oxidation process, the oxidation potentials shifted to a lower potential with an increase in the number of methyl substituents on the ruthenocenyl ring, and shifted to a higher potential with the increase in the number of C[triple bond]C units; this result is in contrast to that found in the [Rc(CH=CH)(n)Rc] series. The chemical oxidation of [Rc'C[triple bond]CRc'] yielded a stable two-electron-oxidized species, the structure of which was confirmed by X-ray crystallography to be [Ru2(mu2-eta(6):eta(6) C5Me4C=CC5Me4)(eta-C5H5)2](BF4)2. Changing the substituents (Rc, Rc*, and Rc') had no effect on the chemical oxidation, but in the case of the Rc' series the Me substituent increased the stability of the two-electron-oxidized species in solution. The diyne [Rc*(C[triple bond]C)2Rc*] and the triyne [Rc*(C[triple bond]C)3Rc*] also gave a similar but unstable two-electron-oxidized species. In acetone or acetonitrile, the two-electron-oxidized species of [Rc*C[triple bond]CRc*] and [Rc*(C[triple bond]C)2Rc*] gradually formed the corresponding bis(fulvene)-type complexes. This implies that the two-electron-oxidized species of [Rc*(C[triple bond]C)(n)Rc*] are destabilized with the increasing n. PMID- 16342317 TI - Polar self-assembly: steric effects leading to polar mixed-ligand coordination cages. AB - We present a highly unusual example of self-assembly, specifically a polar, mixed ligand cage which forms in preference to symmetrical homo-ligand products, and which suggests that steric effects can be exploited to obtain novel non-uniform polyhedral cages. In particular, reaction between the bulky tripodal triphosphine 2,4,6-tris(diphenylphosphino)triazine, L1, the non-bulky tripodal trinitrile 2,4,6-tris(cyanomethyl)trimethylbenzene, L2 and silver hexafluoroantimonate, AgSbF6, in a 3:1:4 ratio gives the mixed-ligand aggregate [Ag4(L1)3(L2)(SbF6)]3+, 1-SbF6, instantly as the only product in quantitative yield. The X-ray crystal structure of complex 1-SbF6 is consistent with the suspected solution-state structure. The cage derives from trigonal-pyramidal geometry, the basal vertices of which are defined by three bulky triphosphines, L1, and the apical vertex by the non-bulky trinitrile, L2. There is apical elongation amounting to 19% in comparison to the ideal uniform tetrahedron. The cage also encapsulates an SbF6 anion. 19F NMR spectra in solution for the analogous PF6 complex [Ag4(L1)3(L2)(PF6)]3+, 1-PF6, confirm that one anion is also encapsulated in solution. The synthesis of the analogous CF3SO3(-) complex, [Ag4(L1)3(L2)(OTf)]3+, 1-OTf, in solution is also described, although 1-PF6 and 1 OTf could not be isolated due to slow decomposition in solution. The selective formation of these mixed-ligand cages is discussed in terms of ligand-ligand and ligand-included anion steric repulsions, which we propose prevent the formation of the competing hypothetical homo-ligand tetrahedral structure [Ag4(L1)4(SbF6)]3+, and thus favour the mixed ligand cage. "Cage cone angles" for L1 and L2 are estimated at 115 degrees and 101 degrees, respectively. Variable temperature 31P NMR spectroscopy shows that complex 1-SbF6 and the related previously reported partial tetrahedral complex [Ag4(L1)3(anion)]3+ undergo dynamic twisting processes in solution between enantiomeric C3-symmetry conformations. PMID- 16342318 TI - Protein function microarrays based on self-immobilizing and self-labeling fusion proteins. AB - Protein microarrays are an attractive approach for the high-throughput analysis of protein function, but their impact on proteomics has been limited by the technical difficulties associated with their generation. Here we demonstrate that fusion proteins of O6-alkylguanine-DNA alkyltransferase (AGT) can be used for the simple and reliable generation of protein microarrays for the analysis of protein function. Important features of the approach are the selectivity of the covalent immobilization; this allows for direct immobilization of proteins out of cell extracts, and the option both to label and to immobilize AGT fusion proteins, which allows for direct screening for protein-protein interactions between different AGT fusion proteins. In addition to the identification of protein protein interactions, AGT-based protein microarrays can be used for the characterization of small molecule-protein interactions or post-translational modifications. The potential of the approach was demonstrated by investigating the post-translational modification of acyl carrier protein (ACP) from E. coli by different phosphopantetheine transferases (PPTases), yielding insights into the role of selected ACP amino acids in the ACP-PPTase interaction. PMID- 16342319 TI - A novel diagnostic strategy for trisomy 21 using short tandem repeats. AB - Molecular technique with STRs can rapidly diagnose aneuploidy. In order to improve its fidelity, we developed a novel STR-based strategy for fast diagnosis of trisomy 21 and constructed a multimarker diagnostic system according to it. The system is based on nine STRs, of which two were previously known and seven were newly identified from the genomic sequence of the long arm of chromosome 21. They were confirmed to be highly polymorphic in the Chinese population by PCR amplification and gel electrophoresis. The combination of nine STR markers, when applied to DNA from 102 Chinese individuals with normal karyotype, did not yield any false-positives, and clearly revealed three different alleles in DNA from 15 out of 18 trisomy 21 patients. The results show that our new strategy can provide an alternative molecular technique for the rapid detection of aneuploidy. PMID- 16342320 TI - Development of a capillary electrophoresis method for the determination of soybean proteins in soybean-rice gluten-free dietary products. AB - CE has been applied for the first time to the simultaneous separation of soybean and rice proteins. Treated and untreated capillaries with different effective lengths as well as separation media at different pHs were tested. For that purpose, samples and standard solutions were prepared in 25:75 ACN-water media containing 0.3% v/v acetic acid. The use of an untreated capillary of 50 cm effective length together with an 80 mM borate buffer (pH 8.5) modified with 20% v/v ACN and UV detection at 254 nm were the conditions working the best. These conditions enabled the determination of soybean proteins in gluten-free dietary commercial products elaborated with soybean protein and/or soybean flour and rice flour using the standard additions calibration method. The method was linear up to 26 mg/mL of soybean proteins, the precision (expressed as RSD) was always better than 6%, and recoveries obtained for soybean proteins when spiking commercial products were very close to 100%. PMID- 16342321 TI - Effect of monosaccharide composition, glycosidic linkage position and anomericity on the electrophoretic mobility of labeled oligosaccharides. AB - Fluorophore-assisted carbohydrate electrophoresis (FACE) is useful for separation and characterization of oligosaccharides from various sources and for comparing several samples at once. While characterizing fungal surface glycans by FACE we observed that samples and standards of the same mass did not comigrate as expected. Subsequent experiments showed that the samples did not contain contaminating sugars. Therefore, our observation suggested that glycan electrophoretic mobility is affected by factors in addition to molecular mass. This work assesses the contribution of monosaccharide composition, linkage position, and linkage anomericity to glycan mobility. Commercially available (and synthesized when available) bioses of known composition were derivatized with a charged fluorophore, and electrophoretic mobilities compared in a slab gel format. The results indicate that all three parameters mentioned above affect observed migration. Further, no migration patterns emerged to suggest a set of rules for assigning band identity based on mobility alone. These results emphasize the importance of including known, matched, standards to facilitate interpretation of FACE data. PMID- 16342322 TI - Ion-pair association and acid-base equilibria in nonaqueous capillary electrophoresis of weakly basic compounds. AB - CE in nonaqueous media was used to study the migrating behavior of two weakly basic s-triazine pesticides and one of their metabolites. The target pesticides were selected to be representative for each of the two main groups: propazine and deethylatrazine for the chloro-s-triazines group and ametryn for the methylthio-s triazines group. To elucidate the phenomena involved, systematic studies were carried out in the different organic media studied. Absolute mobilities were determined in 50% v/v methanol (MeOH)/ACN by extrapolation of the effective mobilities to zero ionic strength in the presence of different concentrations of perchloric acid. Conductivity measurements performed in MeOH and 50 and 20% v/v methanol/ACN permitted the evaluation of the associations of the components of the BGE. The effects of ionic strength on the actual mobilities of the compounds were determined in the presence of perchloric acid and SDS in different organic media. Two different ion-pair equilibria were considered: one due to the presence of perchlorate anions present in the BGE and second that from the added dodecyl sulfate anions. Bearing in mind that these weakly basic compounds can exhibit ion pair and acid-base equilibria, the acid-base and ion-pair parasite reaction coefficients were determined. Finally, the effects of ionic strength, ion-pair interactions and acid-base properties on the effective electrophoretic mobilities of the analytes are discussed. PMID- 16342323 TI - Capillary electrophoresis versus differential scanning calorimetry for the analysis of free enzyme versus enzyme-ligand complexes: in the search of the ligand-free status of cholinesterases. AB - Cholinesterases (ChEs) are highly efficient biocatalysts whose active site is buried in a deep, narrow gorge. The talent of CE to discover inhibitors in the gorge of highly purified preparations has fairly altered the meaning of a ChE ligand-free status. To attempt at a description of this one, we investigated the stability of Bungarus fasciatus acetylcholinesterase (AChE), alone or complexed with different inhibitors. Determination of mid-transition temperature for thermal denaturation, using differential scanning calorimetry (DSC) and CE, provided conflicting results. Discrepancies strongly question the reality of a ligand-free AChE state. DSC allowed estimation of the stability of AChE-ligands complexes, and to rank the stabilizing effect of different inhibitors. CE acted as a detector of hidden ligands, provided that they were charged, reversibly bound, and thus dissociable upon action of electric fields. Then, CE allowed quantification of the stability of ligand-free AChE. CE and DSC providing each fractional and nonredundant information, cautious attention must be paid for actual estimation of the conformational stability of ChEs. Because inhibitors used in purification of ChEs by affinity chromatography are charged, CE remains a leading method to estimate enzyme stability and detect the presence of bound hidden ligands. PMID- 16342324 TI - An inexpensive microslab gel DNA electrophoresis system with real-time fluorescence detection. AB - In this paper, we describe the construction of a simple yet powerful gel electrophoresis apparatus that can be used to perform size-selective separations of DNA fragments in virtually any laboratory. This system employs a microslab gel format with a novel gel casting technique that eliminates the need for delicate combs to define sample loading wells. The compact size of the microslab gel format allows rapid separations to be performed at low voltages using submicroliter sample volumes. Real time fluorescence detection of the migrating DNA fragments is accomplished using an inexpensive digital microscope that directly connects to any PC with a USB interface. The microscope is readily adaptable for this application by replacing its white light source with a blue light-emitting diode (LED) and adding an appropriate emission filter. Both polyacrylamide and agarose gels can be used as separation matrices. Separation performance was characterized using standard dsDNA ladders, and correct sizing of a 191 bp PCR product was achieved in 15 min. The low cost and simplicity of this system makes it ideally suited for use in a variety of laboratory and educational settings. PMID- 16342325 TI - Serum and mucosal antibody responses to pneumococcal protein antigens in children: relationships with carriage status. AB - Streptococcus pneumoniae causes significant morbidity and mortality especially in children. Some pneumococcal protein antigens can protect mice against infection. Little information is available concerning the nature of naturally acquired protective immunity to pneumococci in humans induced by these antigens. This study investigates the relationships between systemic and local antibody production and carriage in children. Children undergoing adenoidectomy (n=112, ages 2-12 years) were studied. Nasopharyngeal swabs were collected for pneumococcal culture. Serum and saliva were assayed for antibodies to several pneumococcal proteins: choline binding protein A (CbpA), pneumolysin (Ply), pneumococcal surface adhesin A (PsaA) and pneumococcal surface protein A (PspA). Adenoidal mononuclear cells (MNC) were cultured with pneumococcal culture supernatants or recombinant proteins. Cell culture supernatants were analyzed for antigen-specific antibodies. Carriage rates fell with age and serum levels of anti-CbpA, Ply and PspA rose. Anti-CbpA and -Ply serum and salivary IgG antibody levels were higher in children who were culture negative than those who were colonized. Antigen stimulation increased respective antigen-specific IgG production by adenoidal MNC and these responses were greater in those who were colonized than in culture-negative children. Antibodies to CbpA and Ply may protect children aged 2 years and older against pneumococcal colonization. Adenoids may be important local induction and effector sites for both mucosal and systemic antibody production to pneumococcal proteins in children. PMID- 16342326 TI - CD69 down-modulation and inhibition of thymic egress by short- and long-term selective chemical agonism of sphingosine 1-phosphate receptors. AB - Thymic development requires proliferation, selection, maturation and release of mature single-positive CD4 and CD8 T cells into the periphery. In mice, non selective sphingosine-1 phosphate (S1P) receptor agonists, active on four of the five known S1P receptors, alter thymocyte phenotype and egress. Here, we show that down-modulation of CD69 occurs acutely and transiently at a discrete and late stage of medullary development after a single-dose administration of S1P(1) receptor-selective agonist, which induces long-term tonic receptor activation in the absence of receptor degradation. In addition, agonist acutely inhibited egress of mature thymocytes into peripheral lymphoid organs, suggesting that both the phenotype and migration of medullary thymocytes are regulated simultaneously and coordinately by agonism of S1P(1) alone. Long-term dosing shifted the early/late medullary thymocyte ratio with an expansion of the late medullary compartment, as mature CD69(-) thymocytes were retained within the thymus. Therefore, chemical agonism of S1P(1) accelerates medullary phenotypic maturation and inhibits egress, leading to the expansion and accumulation of the recent thymocyte emigrant population in the medulla. However, chemical agonism fails to replicate the S1P(1)-null CD69(hi) late medullary phenotype, suggesting that agonism and gene deletion operate by distinct mechanisms, and that functional receptor antagonism may not be required for lymphocyte sequestration. PMID- 16342327 TI - The immunomodulator ginsan induces resistance to experimental sepsis by inhibiting Toll-like receptor-mediated inflammatory signals. AB - Ginsan, a polysaccharide extracted from Panax ginseng, has multiple immunomodulatory effects. In this study, we show that pretreatment of ginsan (25 mug/kg) protected mice from lethality induced by Staphylococcus aureus challenge. This survival benefit was associated with enhanced bacterial clearance from circulation, spleen and kidney. The phagocytic activity of macrophages treated with ginsan was significantly enhanced against S. aureus. However, the production of proinflammatory cytokines, such as TNF-alpha, IL-1beta, IL-6, IFN-gamma, IL 12, and IL-18, was markedly down-regulated in ginsan-treated mice compared with those of control-infected mice. The expression of Toll-like receptor (TLR) 2 and the adaptor molecule MyD88, which was greatly increased in septic macrophages, was significantly reduced by ginsan treatment in vitro. Similarly, the expression of phospho-JNK1/2, phospho-p38 MAPK, and NF-kappaB was decreased in the same culture system. These results illustrate that the antiseptic activity of ginsan can be attributed to enhanced bacterial clearance, and reduced proinflammatory cytokines via the TLR signaling pathway. PMID- 16342328 TI - The chicken Ig light chain 3'-enhancer is essential for gene expression and regulates gene conversion via the transcription factor E2A. AB - Expression of the rearranged chicken immunoglobulin light chain (IgL) gene is regulated by a V gene promoter, a matrix attachment region (MAR) in the J-C intron and an enhancer downstream of the Ig constant region. Using knockout analysis, we demonstrate that the 3'-enhancer is not only required for gene activation but is also essential for the maintenance of gene expression. Deletion of the MAR on the other hand increases IgL transcription, indicating that the MAR acts as negative regulator. We demonstrate that Id1 and Id3, dominant-negative regulators of basic-region helix-loop-helix (bHLH) transcription factors, are able to reduce chicken IgL 3'-enhancer activity in transient assays and strongly reduce the rate of gene conversion (GC) in DT40 clone 18 cells. Conversely, overexpression of avian E47, a bHLH transcription factor, leads to a dramatic increase in GC rates independent of IgL or activation-induced cytidine deaminase RNA levels. Thus, E47 is the first transcription factor to activate GC without an apparent increase in transcription. PMID- 16342331 TI - Solution, solid phase and computational structures of apicidin and its backbone reduced analogs. AB - The recently isolated broad-spectrum antiparasitic apicidin (1) is one of the few naturally occurring cyclic tetrapeptides (CTP). Depending on the solvent, the backbone of 1 exhibits two gamma-turns (in CH(2)Cl(2)) or a beta-turn (in DMSO), differing solely in the rotation of the plane of one of the amide bonds. In the X ray crystal structure, the peptidic C==Os and NHs are on opposite sides of the backbone plane, giving rise to infinite stacks of cyclotetrapeptides connected by three intermolecular hydrogen bonds between the backbones. Conformational searches (Amber force field) on a truncated model system of 1 confirm all three backbone conformations to be low-energy states. The previously synthesized analogs of 1 containing a reduced amide bond exhibit the same backbone conformation as 1 in DMSO, which is confirmed further by the X-ray crystal structure of a model system of the desoxy analogs of 1. This similarity helps in explaining why the desoxy analogs retain some of the antiprotozoal activities of apicidin. The backbone-reduction approach designed to facilitate the cyclization step of the acyclic precursors of the CTPs seems to retain the conformational preferences of the parent peptide backbone. PMID- 16342332 TI - Site-specific synthesis of Amadori-modified peptides on solid phase. AB - Glycation of peptides and proteins is a slow chemical reaction of reducing sugars modifying the amino groups. The first intermediates of this nonenzymatic glycosylation are the Amadori products that can undergo further chemical reactions, finally leading to advanced glycation end products (AGEs). The formation of AGEs was not only linked to aging of tissues and organs in general but also to several diseases such as diabetes mellitus and Alzheimer's disease. Because of the importance of these modifications and their potential use as diagnostic markers, a global postsynthetic approach on solid phase was developed. The peptides were synthesized by Fmoc/(t)Bu-chemistry, with the lysine residue to be modified being protected with the very acid-labile methyltrityl group. Incubation of the peptides with D-glucose in DMF at elevated temperatures resulted in product yields of 35%. Neighboring residues with bulky protecting groups reduced the yields only slightly. The major by-products were the unmodified peptide and an oxidation product. Whereas the unmodified peptide eluted before the glycated peptide, all other by-products eluted later in RP HPLC, allowing simple purification. PMID- 16342333 TI - Influence of vascular filling and perfusion on BOLD contrast during reactive hyperemia in human skeletal muscle. AB - Mechanisms generating BOLD contrast are complex and depend on parameters that are prone to large variations, in particular in skeletal muscle. Here, we simultaneously measured perfusion by ASL, and BOLD response in the calf muscle of 6 healthy volunteers during post-ischemic reactive hyperemia. We tested whether the relation between the two was altered for varying degrees of leg vascular replenishment induced by prior positioning of the leg at different heights relative to the heart. We found that the BOLD response depended on perfusion, but also on the degree of repletion of leg blood vessels. We conclude that simultaneous determination of perfusion by ASL is important to identify the mechanisms underlying BOLD contrast in the skeletal muscle. PMID- 16342334 TI - The hydroperoxy radical as a hydrogen bond acceptor. HOO-HCl complexes--Ab initio study. AB - Protonacceptor properties of the HOO radical were investigated previously by means of ab initio as well as topological Atoms in Molecules (AIM) and Electron Localization Function (ELF) methods. It was pointed out that in the radical there are three nonequivalent positions most susceptible to protonation, and on this basis three structures of possible hydrogen bonded complexes were proposed. Results reported in the present article concern all possible 1:1 complexes formed by HCl and HOO molecules, and fully confirm suppositions given on the basis of the above-mentioned investigations. There are three various complexes referring to the local minima, and the transition structure predicted by topological methods has been found as well. The cyclic structure appeared to be the most stable one, which confirms conclusions given in the experimental article. Apart from structure optimization, harmonic as well as anharmonic spectra of the complexes have been simulated. Anharmonicity of H-Cl stretching vibration was of special interest, as the frequency of this vibration characterizes the Cl-H...O hydrogen bond in these complexes. To obtain values of these frequencies the one dimensional Hamiltonian has been diagonalized numerically. The potential for this Hamiltonian has been taken from a set of single-point scanning of the part of the Potential Energy Surface (PES) connected with this vibration. The potential calculated on the MP2 level leads to the result close to the experimental value, whereas the B3LYP method is inappropriate for the purpose of PES investigation of these complexes. PMID- 16342335 TI - Electron group functions for the analysis of the electronic structures of molecules. AB - The electronic structure of a vast majority of molecular systems can be understood in terms of electron groups and their wave functions. They serve as a natural basis for bringing intuitive chemical and physical concepts into quantum chemical calculations. This article considers the general electron group functions formalism as well as its simple geminal version. We try to characterize the wave function with the group structure and its capabilities in actual calculations. For this purpose we implement a variational method based on the wave function in the form of an antisymmetrized product of strongly orthogonal group functions and perform a series of electronic structure calculations for small molecules and model systems. The most important point studied is the relation between the choice of electron groups and the results obtained. We consider energetic characteristics as well as optimal geometry parameters. In view of practical importance, the structure of variationally optimized local one electron states is considered in detail as well as intuitive characteristics of chemical bonds. PMID- 16342336 TI - Bayesian decision procedures for dose-escalation based on evidence of undesirable events and therapeutic benefit. AB - In this paper, Bayesian decision procedures are developed for dose-escalation studies based on bivariate observations of undesirable events and signs of therapeutic benefit. The methods generalize earlier approaches taking into account only the undesirable outcomes. Logistic regression models are used to model the two responses, which are both assumed to take a binary form. A prior distribution for the unknown model parameters is suggested and an optional safety constraint can be included. Gain functions to be maximized are formulated in terms of accurate estimation of the limits of a 'therapeutic window' or optimal treatment of the next cohort of subjects, although the approach could be applied to achieve any of a wide variety of objectives. The designs introduced are illustrated through simulation and retrospective implementation to a completed dose-escalation study. PMID- 16342338 TI - Cardiovascular diseases in Croatia and other transitional countries: comparative study of publications, clinical interventions, and burden of disease. AB - AIM: To determine the number of publications on cardiovascular diseases in the MEDLINE database, the rate of medical doctors and clinical interventions in cardiology, and health and socioeconomic indicators for Croatia, and to compare them with those for Slovenia, Hungary, the Czech Republic, and Austria. METHODS: PubMed was used in search for publications on cardiovascular diseases published in 1991-2004. Rates per million population and proportions of publications on cardiovascular diseases in the MEDLINE database were calculated. Gross domestic product (GDP) per capita was used as a socioeconomic indicator, whereas human resources in medicine were presented as the rate of medical doctors per million population. Standardized death rates from cardiovascular diseases and ischemic heart disease were used as indicators of cardiovascular health. Clinical interventions in cardiology, such as coronary angiograms, percutaneous transluminal coronary angioplasties (PTCA), and coronary bypass surgeries (CABG) were expressed per million population per year. RESULTS: Croatia had the lowest GDP per capita among the analyzed countries. The standardized death rate from cardiovascular diseases in Croatia was 91.7 per 100,000 population aged 0-64 in 2001, which was higher than that in Slovenia and Austria (P<0.001), similar to that in the Czech Republic, and lower than that in Hungary (P<0.001). Cardiovascular scientific output in Croatia was the lowest among investigated countries, ie, 1.1 per million population in 2003 (P<0.001). Despite a significantly lower number of medical doctors in comparison with Hungary and the Czech Republic (P<0.001), Croatia experienced a similar increment in the amount of clinical interventions in cardiology. CONCLUSION: In contrast to high cardiovascular mortality rates, cardiovascular scientific production in Croatia was significantly lower than in other investigated countries. A positive trend in cardiovascular medicine was recorded in clinical practice, but has yet to be followed by scientific production. PMID- 16342339 TI - Web quality control for lectures: Supercourse and Amazon.com. AB - Peer review has been at the corner stone of quality control of the biomedical journals in the past 300 years. With the emergency of the Internet, new models of quality control and peer review are emerging. However, such models are poorly investigated. We would argue that the popular system of quality control used in Amazon.com offers a way to ensure continuous quality improvement in the area of research communications on the Internet. Such system is providing an interesting alternative to the traditional peer review approaches used in the biomedical journals and challenges the traditional paradigms of scientific publishing. This idea is being explored in the context of Supercourse, a library of 2,350 prevention lectures, shared for free by faculty members from over 150 countries. Supercourse is successfully utilizing quality control approaches that are similar to Amazon.com model. Clearly, the existing approaches and emerging alternatives for quality control in scientific communications needs to be assessed scientifically. Rapid explosion of internet technologies could be leveraged to produce better, more cost effective systems for quality control in the biomedical publications and across all sciences. PMID- 16342340 TI - Myocardial protective effect of warm blood, tepid blood, and cold crystalloid cardioplegia in coronary artery bypass grafting surgery. AB - AIM: To compare the myocardial effects of cardioplegia by warm blood, tepid blood, and cold crystalloid during coronary artery bypass grafting (CABG). METHODS: Patients undergoing CABG surgery at Kaunas University Hospital between 2000 and 2004 were randomized into three groups (n=156), receiving a different method of cardioplegia. Intermittent antegrade warm blood cardioplegia was used in 51 patients, tepid blood cardioplegia in 50 patients, and cold crystalloid cardioplegia in 55 patients. Mitochondrial function, myocardial ultrastructure, troponin T, and hemodynamic and clinical data were analyzed after surgery. RESULTS: All cardioplegic methods similarly affected structural and functional properties of mitochondria and coupling of oxidative phosphorylation, and all lowered the capacity of mitochondria to synthesize ATP. Ultrastructure of myocytes showed slight to moderate injury in the cold crystalloid cardioplegia group. The concentration of troponin T was significantly lower in the warm blood cardioplegia group than in the tepid blood cardioplegia and cold crystalloid cardioplegia groups at 12 hours (0.8+/-0.1 ng/mL, 1.9+/-0.2 ng/mL, and 2.8+/-0.3 ng/mL, respectively; P<0.001) and 24 hours after surgery (1.0+/-0.1 ng/mL, 2.2+/ 0.3 ng/mL, and 2.5+/-0.3 ng/mL, respectively; P<0.001). Echocardiographic examination after surgery revealed that the changes in the left ventricle diastolic function were similar in all groups, and that systolic function did not change. The warm blood cardioplegia group showed shorter duration of intubation and hospitalization. There were no differences in the need of catecholamine administration, incidence of complications, and duration of stay in the intensive care unit. CONCLUSIONS: Intermittent antegrade warm blood cardioplegia provides better myocardial protection during CABG surgery, as assessed by the lower release of troponin T, lower fluid balance, shorter duration of tracheal intubation and hospital stay. PMID- 16342341 TI - Decreased nitric oxide in women with essential hypertension in prehypertensive phase. AB - AIM: To determine the concentrations of nitric oxide (NO) in plasma of women with essential hypertension in prehypertensive phase, its effect on blood pressure, and correlation with other vasoactive substances that regulate systemic and renal vascular tonus. METHODS: The study performed at the Department of Nephrology, Hospital Center in Skopje, Macedonia, included 26 women with essential hypertension in prehypertensive phase and 11 normotensive women as healthy controls. Vasodilating factors NO and 6-keto-prostaglandin F1 alpha (6-keto PGF1alpha) were determined in plasma. Thromboxane B2 (TXB2) as a vasoconstricting factor and electrolytes Na+, K+, and Ca2+ were determined in urine. Blood pressure was monitored over 24 hours. Systolic, diastolic, mean blood pressure were presented as average 24-hour values. RESULTS: The concentrations of NO and 6 keto-PGF1alpha were significantly lower in women with essential hypertension in prehypertensive phase than in their normotensive controls (NO: median 22, range 11-35 vs median 37.5, range 11-66; 6-keto-PGF1alpha: 64.8+/-14.35 vs 98.21+/ 43.45 micromol/L; P<0.001). The index of vascular reactivity (TXB2/6-keto PGF1alpha ratio) was higher in women in prehypertensive phase than in normotensive women (1.3 vs 0.8, P<0.001). Urinary calcium to creatinine ratio was significantly lower in the prehypertensive group (0.06+/-0.03 vs 0.24+/-0.13, P<0.001). No direct correlations were found between NO, TXB2, and 6-keto PGF1alpha, or between NO and electrolytes in the urine. Low NO and urinary Ca2+ were significant indicators of increased blood pressure (P=0.013 and P=0.024, respectively; backward stepwise multiple regression analysis). CONCLUSIONS: NO and 6-keto-PGF1alpha were significantly lower in women in prehypertensive phase of essential hypertension. Lower NO correlated with increased systolic blood pressure, but not with on natriuresis and calciuresis. These findings, together with the higher vascular reactivity index, indicate that endothelial dysfunction precedes the establishment of essential hypertension. PMID- 16342342 TI - Triggering of ventricular ectopic beats by emotional, physical, and meteorologic stress: role of age, sex, medications, and chronic risk factors. AB - AIM: To investigate the association of a single ventricular ectopic beat with physical, emotional, or meteorologic stress and the role of age, sex, antiarrhythmic and other medicament therapy and participant characteristics. METHODS: The study included 457 participants who were consecutively assigned to undergo continuous 24-hour Holter monitoring and who completed a structured questionnaire about their physical activity and mental stress. Multiple regression analysis of data on 11 meteorologic parameters, participants' baseline characteristics, and medications they used was performed for subgroups according to sex and age, with 65 years as age limit. RESULTS: Wind speed, low relative humidity, increasing relative humidity, and emotional stress were independent predictors of ventricular ectopic beat, whereas warm front passage showed a protective effect in all participant subgroups (P<0.05 in all cases). Physical activity was an independent predictor of ventricular ectopic beat in women (P=0.02) and cold front passage in men P=0.003). The circadian variation in frequency of ventricular ectopic beats persisted in all subgroups after adjustments for external triggers (P<0.001 in all cases). Among chronic risk factors, familial background and previous myocardial infarction were independent predictors of ventricular ectopic beat in men and younger (P<0.05 in all cases). Nitrates (P<0.03 in all cases) and angiotensin converting enzyme inhibitors (P<0.02 in all cases) exerted protective effect in younger participants as well as did beta-blockers in the elderly (P<0.003 in all cases). CONCLUSION: In addition to existence of an endogenous, external triggering-independent circadian pattern, physical, emotional, and meteorologic stress may act as triggers of ventricular ectopic beat in a manner that differs from triggering more complex arrhythmias. Familial background seems to have a significant impact on arrhythmogenesis in men. PMID- 16342343 TI - In-hospital cardiac arrest and resuscitation outcomes: rationale for sudden cardiac death approach. AB - AIM: To assess the frequency of cardiac arrest and outcomes and predictors of survival after cardiopulmonary resuscitation in hospitalized patients. METHODS: We prospectively analyzed the data on all patients who experienced cardiac arrest while hospitalized at the Split University Hospital between January and December 2003. Data were collected on patients' demographic characteristics, etiology and presentation of cardiac arrest, time, site, methods, and outcomes of cardiopulmonary resuscitation. RESULTS: Out of 120 cases of cardiac arrest among 32,861 hospitalized patients, 76.7% were witnessed. Ninety-six (80.0%) patients with cardiac arrest underwent resuscitation, and 22.5% of them were discharged alive. The survival rate was 20.0% at the Department of Internal Medicine, 29.2% in the Coronary Care Unit, and only 7.1% in other departments (P=0.058, chi2 test). Out of 92 patients with witnessed cardiac arrest, 28.3% survived to discharge, whereas only one of 28 patient with unwitnessed cardiac arrest survived to discharge (P=0.004, Fisher's exact test). More patients with cardiac arrest due to ventricular fibrillation or pulseless ventricular tachycardia survived than patients with asystole and pulseless electrical activity (47.6% vs 10.7%, respectively, P<0.001, Fisher's exact test). None of the patients with unclassified cardiac arrest survived until discharge. Cardiac arrest survivors were significantly younger (60.8+/-12.9 vs 71.1+/-11.7 years, P<0.001, Student t test). Sex had no influence on survival. There were no significant circadian or hospital shift differences in the frequency rate of cardiac arrest, but the rate of successful resuscitation was lower during the night shift. CONCLUSION: The rate of successful resuscitation was higher in the coronary care unit, during the day and in younger witnessed cardiac arrest patients with ventricular fibrillation or pulseless ventricular tachycardia. PMID- 16342344 TI - Early detection of left ventricular diastolic dysfunction in hypertensive heart disease by color Doppler myocardial imaging. AB - AIM: To determine if Color Doppler myocardial imaging could provide evidence of diastolic dysfunction in patients with hypertension whose pulse-wave Doppler parameters were normal. METHOD: The study included 33 patients (mean age 48+/-7.3 years) and a control group of 13 sex- and age-matched healthy individuals. Patients were divided into two groups according to mean blood pressure (BP) values during 24-hour blood pressure monitoring while under antihypertensive therapy: those with uncontrolled hypertension (n=22) and those with controlled hypertension (n=11). All study participants underwent complete standard echocardiography (2D, M-mode, pulsed and continuous Doppler) and a Color Doppler myocardial imaging study. RESULTS: Conventional Doppler parameters indicated relaxation disturbances in patients with uncontrolled hypertension, but were within a normal range in patients with controlled hypertension at baseline and follow-up. Parameters of global diastolic function measured by Color Doppler myocardial imaging revealed that E'/A', the ratio between E'-wave (early filling phase) and A'-wave (late diastolic wave due to atrial contraction), was <1 in 57% of segments at baseline in patients with uncontrolled hypertension, and did not significantly change at follow-up. In patients with controlled hypertension, E'/A'<1 was noted in 4.7% of segments at baseline and in 28.6% of segments at follow-up. CONCLUSION: Regional diastolic dysfunction measured by Color Doppler myocardial imaging was the first sign of myocardial dysfunction due to arterial hypertension, while the parameters of global diastolic dysfunction measured by conventional Doppler and Color Doppler myocardial imaging were still normal. Furthermore, in patients with uncontrolled hypertension with manifested global diastolic dysfunction, there was a change in late diastolic parameters. Our results point to a potentially important role of Color Doppler myocardial imaging in diagnosing hypertensive heart disease as well as in follow-up of treatment. PMID- 16342345 TI - Ventricular pacing vs dual chamber pacing in patients with persistent atrial fibrillation after atrioventricular node ablation: open randomized study. AB - AIM: To compare ventricular rate responsive (VVIR) pacing with dual chamber rate responsive (DDDR) pacing and antiarrhythmic drugs for the treatment of patients with persistent atrial fibrillation after atrioventricular node ablation. METHODS: One hundred two patients with persistent atrial fibrillation eligible for the atrioventricular node ablation were randomly assigned to the therapy with either VVIR pacemaker (n=52) or DDDR pacemaker and antiarrhythmic drugs (n=50). After ablation, patients in both pacing groups were assigned to take anticoagulant therapy. The primary end point was stroke or death from cardiovascular causes. RESULTS: After a mean follow-up of 26.6+/-9.5 months, there was no difference in the stroke or death rates between patients with VVIR pacing (6 or 5.2% per year) and those with DDDR pacing and antiarrhythmic drugs (6 or 5.9% per year, P=0.930). The observed rates of death from all causes, hospitalization for heart failure, and myocardial ischemia were similar in the two patient groups. There was a significantly lower rate of development of permanent atrial fibrillation in patients with DDDR pacing and antiarrhythmic drugs, with a reduction in absolute risk by 56% and relative risk by 64% (32% vs 88%; P<0.001). CONCLUSION: With respect to cardiovascular death and morbidity, VVIR pacing is not inferior to DDDR pacing and antiarrhythmic drugs for the treatment of patients with persistent atrial fibrillation after atrioventricular node ablation and may be considered as an appropriate therapy for such patients. PMID- 16342346 TI - Impact of atrioventricular node ablation and pacing therapy on clinical course in patients with permanent atrial fibrillation and unstable ventricular tachycardia induced by rapid ventricular response: follow-up study. AB - AIM: To evaluate prospectively the impact of atrioventricular (AV) node ablation and consequent pacing therapy on clinical course in patients with permanent atrial fibrillation and unstable ventricular tachycardia induced by rapid ventricular response. METHODS: One hundred four patients with permanent atrial fibrillation and uncontrolled ventricular rate resistant to drug therapy underwent radiofrequency catheter ablation of the AV node and permanent pacemaker implantation. At baseline examination, 14 of them had unstable ventricular tachycardia induced by rapid ventricular response of atrial fibrillation (ventricular tachycardia group). The remaining 90 patients did not have this type of ventricular tachycardia (control group). After the ablation, all patients were followed-up without antiarrhythmic agents. The primary end point was sudden cardiac death. RESULTS: Before the ablation, patients in ventricular tachycardia group had lower left ventricular ejection fraction (P<0.013), and higher ventricular rate at rest and during daily activities (P<0.001). During the follow up of 20+/-8 months (mean+/-standard deviation), the mortality rate of sudden cardiac death at two years was similar among the two groups (7% vs 5%, P=0.703). The observed cardiac and all-cause mortality were significantly higher in ventricular tachycardia group (21% vs 3.6%, P=0.014; 28.5% vs 4.4%, P=0.038; respectively) due to increased heart failure-related mortality (P=0.013). CONCLUSION: In patients with permanent atrial fibrillation and ventricular tachycardia induced by rapid ventricular response, AV node ablation and pacing therapy have beneficial effect in the elimination of this arrhythmia. It seems that these patients do not need a cardioverter defibrillator therapy. PMID- 16342347 TI - Cardiovascular risk factors and diseases strongly predict hemodialysis treatment outcome in maintenance hemodialysis patients. AB - AIM: To identify independent predictors of hemodialysis treatment outcome among major cardiovascular risk factors and pre-existent cardiovascular diseases in maintenance hemodialysis patients with the end-stage renal disease. METHODS: A total of 144 prevalent patients on maintenance hemodialysis at Rijeka University Hospital between 1998 and 2003 were included in the prospective clinical study. Pre-existent cardiovascular risk factors and diseases were identified, as well as their relation to hemodialysis treatment outcome. Primary outcome measure was death, and secondary outcome measure was the length of time from the beginning of patient's hemodialysis treatment to the end of follow-up, ie, end of the study or patient's death. The independent variables on hemodialysis treatment outcome were identified with the multiple linear regressions. RESULTS: Cardiovascular diseases were the major cause of death in 40 (60.6%) patients. Acute myocardial infarction in 15 (22.7%) patients was the major single cause of death. Among risk factors, hyperglycemia (P<0.001), low delivered dialysis dose (P<0.001), use of semi synthetic dialysis membrane (P<0.001), and anemia (P=0.041) were independent predictors of hemodialysis treatment outcome. Hypertensive heart disease (P<0.001), ischemic heart disease (P<0.001), and dilated cardiomyopathy (P=0.016) were independent predictors of the hemodialysis treatment outcome. CONCLUSIONS: Cardiovascular diseases were the leading cause of death in hemodialysis patients. There was also high prevalence of cardiovascular risk factors and pre-existent cardiovascular diseases. Several of them were independent predictors of the hemodialysis treatment outcome. PMID- 16342348 TI - Transient left ventricular apical ballooning mimicking acute coronary syndrome in four patients from central Europe. AB - The syndrome of "apical ballooning" consists of an acute onset of transient extensive akinesia of the apical portion of the left ventricle, without significant stenosis on the coronary angiogram. The syndrome is accompanied by chest symptoms, electocardiographic changes, and limited release of cardiac markers disproportionate to the extent of akinesia. So far, the vast majority of cases with this syndrome have been reported among Japanese population and only a few cases among Caucasian population. We describe "apical ballooning" in four Caucasian patients, three women and one man, who presented at a tertiary referral center over a period of eight months. Their age ranged between 64 and 84 years. Three of them presented with chest symptoms. All four patients had electrocardiographic changes and increased concentration of troponin T. One patient developed hemodynamic instability, but none died or showed recurrence of symptoms during the follow-up of 1-8 months. In all patients, a preceding triggering factor was identified, such as emotional or physical stress. In all patients left ventriculography showed extensive akinesia of the apex of the left ventricle ("apical ballooning") in the absence of a significant coronary artery stenosis. Left ventricular systolic function recovered completely within three days to three weeks. Emotional or physical stress or other preceding triggering factors might play a key role in this cardiomyopathy, but the precise etiology remains unknown. Despite severe initial presentation, conservative medical management leads to good long term outcome. PMID- 16342349 TI - Levosimendan as a new strategy during off-pump coronary artery bypass grafting: double-blind randomized placebo-controlled trial. AB - AIM: To determine if levosimendan could improve cardiac performance during off pump coronary artery bypass grafting in patients with normal preoperative left ventricular function. METHODS: Twenty-four patients included in this double-blind randomized controlled trial received either placebo (n=12) or 12 microg/kg loading dose of levosimendan (n=12) during a period of 15 minutes before the surgery. The heart rate, cardiac index, stroke volume index, and left ventricular ejection fraction were measured before and 10 and 60 minutes after the drug administration. RESULTS: The baseline measurements of hemodynamic parameters did not differ between the groups. Heart rate was not significantly different between the placebo and levosimendan group. Compared with placebo group, cardiac index and left ventricular ejection fraction were significantly higher 10 and 60 minutes (p=0.018 for all) after administration of levosimendan. After 60 minutes, the cardiac index increased from the baseline value of 2.18 to 2.84 L/min/m2. Left ventricular ejection fraction increased by 14% after 10 minutes, and by 10% after 60 minutes. Stroke volume index was significantly higher at 10 minutes (p=0.018), but not at 60 minutes (p=0.063) after the administration of levosimendan. CONCLUSION: Administration of 12 microg/kg of levosimendan enhances the left ventricular performance and offers a promising therapeutic option during off-pump coronary artery bypass grafting in patients with good preoperative left ventricular function. PMID- 16342350 TI - Comparison of on-demand vs planned relaparotomy for treatment of severe intra abdominal infections. AB - AIM: To compare morbidity and mortality of patients with severe intra-abdominal infections after two types of surgical treatment, on-demand ("wait and see") relaparotomy and modified planned relaparotomy. METHODS: We prospectively analyzed the outcomes of 65 patients with severe peritonitis surgically treated in two Croatian hospitals. In one hospital, 34 patients were treated on-demand, and in another hospital 31 patients were treated by planned relaparotomy. We compared severe postoperative complications, mortality, and length of hospital stay in the two groups of patients. RESULTS: Severity of patient's disease, as measured from preoperative group-average Acute Physiology and Chronic Health Evaluation (APACHE) II scores, was comparable in both on-demand and planned relaparotomy groups. The mortality rate was higher in patients operated on-demand (59% vs 29%, P=0.024). In nonadjusted model, the relative risk of dying was 2.5 fold higher for patients treated by on-demand operation in comparison with planned relaparatomy (P=0.030). However, after the adjustment of the survival data for individual patient's sex and APACHE II scores, the difference in the relative risk became non-significant (P=0.178). The patients who died had higher APACHE II scores (26.1+/-8.9 vs 19.7+/-5.9, P=0.009). Relative risk of dying per 5-point increase in APACHE II score was 1.24 (95% confidence interval, 1.01-1.51; P=0.039), irrespective of the surgical technique. CONCLUSIONS: Patients with severe peritonitis treated with planned relaparotomy seemed to have lower mortality. However, the relative risk of dying was not statistically different between the on-demand and planned relaparotomy groups after adjustment for preoperative APACHE II scores. The severity of disease rather than surgical approach plays more important role in survival of these patients. PMID- 16342352 TI - Trends in acute myocardial infarction mortality and morbidity from 1979 to 2001 in the City of Zagreb, Croatia. AB - AIM: To define trends in age-adjusted acute myocardial infarction mortality and morbidity in women and men in the city of Zagreb, Croatia, in the period 1979 2001. METHODS: Acute myocardial infarctions occurring in both men and women at ages between 25 and 74 years were analyzed by using 1979-2001 data from the Acute Myocardial Infarction Population-based Register for the City of Zagreb, Croatia. The patients with myocardial infarction were identified retrospectively from the hospital discharge reports and death certificates in Zagreb hospitals and the Croatian Statistical Bureau. RESULTS: Over the period of 23 years, a total of 25,359 cases of acute myocardial infarction were registered in the city of Zagreb (18,345 men and 7,014 women). The age-adjusted attack rate for men was the lowest in 1981 (170 per 100,000 population) and the highest in 1993 (274 per 100,000 population), with a decreasing trend toward 2001. The rate for women was between 49 per 100,000 population in 1979 and 86 per 100,000 population in 1993, with a decreasing trend afterwards. Between 1993 and 2001, the rate for men decreased by 68.6% and for women by 62.8% (P=0.370). The age-adjusted mortality rate showed more pronounced changes and was much higher for men then for women. The rates per 100,000 population among men varied between 80 and 140, and among women between 20 and 40. Between 1993 and 2001, the age-adjusted myocardial infarction mortality rates decreased by 56.3% and 52.5% in men and women, respectively (P=0.670). The age-adjusted prehospital mortality rate for men was much higher than the hospital mortality rate and had an obvious decreasing trend since 1993. Both rates were much lower for women. Twenty-eight-day case-fatality rate for men ranged from 38.5% to 49.1%, and from 39.0% to 64.0% for women. It did not change much in men during the observed period. Women younger than 45 years had a significantly greater risk of a fatal heart attack, whereas the risk was greater for men in the older age groups (55-74 years). CONCLUSION: The age-adjusted attack rates, mortality rates, prehospital death rates, and case-fatality rates showed a definite decline since the early 1990s for both men and women, but the decline was greater for men. PMID- 16342351 TI - Evaluation of single intensive care unit performance by simplified acute physiology score II system. AB - AIM: To evaluate effectiveness and quality of care in a single intensive care unit (ICU) by the Simplified Acute Physiology Score II (SAPS II). METHODS: A prospective study included 395 patients from the ICU at Rijeka University Hospital, Croatia. The sum of the SAPS II points was used for calculating predicted mortality for each patient. The observed death rate was compared with predicted mortality calculated by SAPS II system. The ability of the SAPS II prognostic system to predict probability of hospital mortality was assessed with discrimination (receiver operating characteristic [ROC] curve) and calibration (Hosmer-Lemeshow test) measures. RESULTS: The SAPS II score on the first ICU day was low (median, 20; range, 3-83). SAPS II system showed a good ability to separate the patients predicted to live from those predicted to die, as shown by an area under the ROC curve of 0.827. The calibration curve demonstrated under prediction of the actual death rate (Hosmer-Lemeshow goodness-of-fit test, C=22.961; df=8; P=0.003). The observed mortality was higher than predicted (observed-to-predicted ratio was 1.6). CONCLUSIONS: SAPS II system is a useful tool for the assessment of ICU performance. This system demonstrated a good ability of discrimination, but an under-prediction of the actual mortality rate in our ICU. PMID- 16342353 TI - Myocardial infarction and religion: hospital-based case-control study in Tirana, Albania. AB - AIM: To assess the association between religion, Muslim vs Christian, and myocardial infarction in the population of Tirana, capital of Albania. METHOD: A hospital-based case-control study included all patients with acute non-fatal myocardial infarction hospitalized in Tirana University Hospital Center between October 1999 and June 2001. There were 146 women with the mean age (+/-standard deviation) of 62+/-9 years and 331 men aged 59+/-10 years. Control group consisted of 616 hospital-based patients recruited from University Departments of Orthopedics and Ear, Nose, and Throat. The control group included 227 women aged 60+/-10 years, and 389 men aged 60+/-9 years. A structured questionnaire was administered to all participants to collect self-reported data on socio demographic factors, life-style and behavioral factors, diet, and pre-existing conditions. All participants had their height and weight measured. RESULTS: Among patients with myocardial infarction, 98 women (67%) and 244 men (74%) identified themselves as Muslims, as compared to 164 women (72%) and 265 men (68%) in the control group. There was no evidence of association between myocardial infarction and religion in women, whereas Muslim men had a higher risk of a first non-fatal myocardial infarction than Christians (age-adjusted odds ratio [OR], 1.32; 95% confidence interval [CI], 0.95-1.82). Upon adjustment for covariates, there was no evidence of a significant association between religion and myocardial infarction neither in men (OR,1.20; 95% CI, 0.85-1.70), nor in women (OR, 0.97; 95% CI, 0.59-1.60). CONCLUSION: The occurrence of myocardial infarction among Muslims and Christians in Tirana was similar, suggesting that cardiovascular morbidity is not affected by the religious affiliation of Albanian adults. PMID- 16342354 TI - Drug therapy of cardiovascular risk factors: guidelines versus reality in primary health care service. AB - AIM: To determine the monthly costs of prescription drugs for the treatment of hypertension, hypercholesterolemia, and diabetes mellitus and their share in the total prescription drugs expenditures allowed by the Croatian Institute for Health Insurance. METHODOLOGY: Hypertension, hypercholesterolemia, and diabetes mellitus were diagnosed in 4,916 patients in three general practitioner (GP) offices in the Zagreb area in January 2005. The monthly cost of medications prescribed by the GPs for the treatment of these three diseases was obtained by summarizing the prices of all antihypertensive, lipid-lowering, and antidiabetic drugs prescribed in one month. The monthly medication costs for each disease separately and all three diseases together were compared with the total prescription drug expenditures approved by the Croatian Institute for Health Insurance. RESULTS: Hypertension was diagnosed in 1,112 (22.6%), hypercholesterolemia in 324 (6.5%), and diabetes mellitus in 359 (7.3%) patients. Monthly cost of all medications prescribed for these three diseases accounted for 81.2% of the total amount approved for medications by the Croatian Institute for Health Insurance. Antihypertensive drugs accounted for 51.1%, antidiabetic drugs for 11.7% for, and lipid-lowering treatment for 18.4%. The medications were prescribed most often to patients older than 65 years (93.6%). CONCLUSION: GPs are strongly obligated by the annual contract with the Croatian Institute for Health Insurance to keep the medication expenses within the approved amount. However, the approved amount is spent mostly on the drug treatment of hypertension, hypercholesterolemia, and diabetes mellitus as the main cardiovascular risk factors. PMID- 16342355 TI - Morbidity of native, immigrant, and returned refugee populations in family medicine practice in Croatia after 1991-1995 war. AB - AIM: To examine the differences in the morbidity among 325 native inhabitants, 231 immigrants, and 145 returned refugees in Komarevo, Croatia. METHODS: The data on patients older than 45 years and their chronic diseases classified according to the 10th revision of International Classification of Diseases and Related Health Problems were collected from the medical files in a family practice in Komarevo. The patients were grouped according to their status (natives, immigrants, and returned refugees) and age (45-64 and > or =65 years). The differences in diagnoses between the groups of the same age were analyzed by descriptive statistics, Kruskal-Wallis test, and chi2 test. RESULTS: In the 45-64 age group, hypertension was diagnosed in 67 out of 183 (36.6%) natives, 22 out of 108 (20.4%) immigrants, and 12 out of 50 (24%) returned refugees (chi2(2)=9.48; P=0.008). In the same age group, ischemic heart disease was found in 21 out of 183 (11.4%) natives, 3 out of 108 (2.8%) immigrants, and 3 out of 50 (6.4%) returned refugees (chi2(2)=7.34, P=0.025). In those aged > or =65, intervertebral disc disorders, dorsalgia, and dorsopathy were found in 61 out of 169 (36.1%) natives, 26 out of 123 (21.1%) immigrants, and 15 out of 90 (15.8%) returned refugees (chi2(2)=15.44, P<0.001). Ischemic heart disease was found in 20 out of 169 (11.8%) natives, 4 out of 123 (3.3%) immigrants, and 16 out of 90 (16.8%) returned refugees (chi2(2)=11.40, P=0.003). Five (56%) and six (67%) out of nine groups of diagnoses had the lowest prevalence in immigrants aged 45-64 and > or =65, respectively. CONCLUSION: Native inhabitants had the highest prevalence of observed chronic diseases, whereas the immigrant adult population had the lowest. Such differences should be kept in mind in the approach to war-affected populations. PMID- 16342356 TI - Wildlife forensic: step forward towards nature protection. PMID- 16342357 TI - Expression of p53 and apoptosis in discoid lupus erythematosus. PMID- 16342358 TI - Cardiac troponin determination in amniotic fluid. PMID- 16342359 TI - Statistical analysis for occupational exposure measurements. PMID- 16342360 TI - After all, that is "science"--the Fifth International Congress on Peer Review and Biomedical Publication, Chicago 2005. PMID- 16342361 TI - Brains in movement. PMID- 16342362 TI - Working for peace through health--ethical values and principles. PMID- 16342363 TI - Five-year experience of Malawi college of medicine with "learning by living" program. PMID- 16342364 TI - Splitting headaches. FTC judge's ruling could have far-ranging impact. PMID- 16342365 TI - [Breast implants and breast hypoplasia or breast augmentation by breast implants]. PMID- 16342366 TI - Re: Iohom G, Fitzgerald D, Cunningham AJ. Principles of pharmacogenetics- implications for the anaesthetist. Br J Anaesth 2004; 93: 440-50. PMID- 16342367 TI - Growing old with HIV. PMID- 16342368 TI - Cerebral palsy: recent insights into movement deviation. PMID- 16342369 TI - State health policy making determinants, theory, and methods: a synthesis. AB - The American states exhibit considerable differences in health policy and market characteristics. Not only do they display substantial variation in spending, but they also display substantial variation in the strategies chosen to control costs, improve access, and ensure quality care. This article synthesizes studies that use 50-state statistical techniques to model policy adoption in the health sector. The purpose is to assess the strengths and weaknesses of this literature, to place it in the context of comparative state policy research generally, and to identify factors that best predict 17 health policy outcomes at the state level. A database was assembled containing 245 equations abstracted from 63 studies published between 1975 and 2002. Some predictors (such as income, aged population, public opinion, and nursing home beds) were studied much more frequently than others (e.g., education, divided government, federal Medicaid mandates, other states' adoptions). Results show that 43 of the 87 policy making determinants examined consistently predict two or more state-level outcomes, including four that predict five outcomes (non-white, urban, income, unemployment), two that predict six (tax capacity/effort, hospital beds), and two that predict seven (nursing home beds, liberal public opinion). Gaps are shown to exist in our understanding of the policy making effects of political system and intergovernmental characteristics. PMID- 16342371 TI - Senate passes amendment to OK HIV Medicaid coverage. PMID- 16342372 TI - Testing. SFAF supports home HIV kits. PMID- 16342370 TI - Mossy fibers are the primary source of afferent input to ectopic granule cells that are born after pilocarpine-induced seizures. AB - Granule cell (GC) neurogenesis increases following seizures, and some newborn GCs develop in abnormal locations within the hilus. These ectopic GCs (EGCs) display robust spontaneous and evoked excitatory activity. However, the pattern of afferent input they receive has not been fully defined. This study used electron microscopic immunolabeling to quantitatively evaluate mossy fiber (MF) input to EGCs since MFs densely innervate the hilus normally and undergo sprouting in many animal models of epilepsy. EGC dendrites were examined in tissue from epileptic rats that had initially been treated with pilocarpine to induce status epilepticus and subsequently had spontaneous seizures. MF terminals were labeled with a zinc transporter-3 antibody, and calbindin immunoreactivity was used to label hilar EGCs and GC layer GCs. The pattern of input provided by sprouted MF terminals to EGC dendrites was then compared to the pattern of MF input to GC dendrites in the inner molecular layer (IML), where most sprouted fibers are thought to project. Analysis of EGC dendrites demonstrated that MF terminals represented their predominant source of afferent input: they comprised 63% of all terminals and, on average, occupied 40% and 29% of the dendritic surface in the dorsal and ventral dentate gyrus, respectively, forming frequent synapses. These measures of connectivity were significantly greater than comparable values for MF innervation of GC dendrites located in the IML of the same tissue sections. Thus, EGCs develop a pattern of synaptic connections that could help explain their previously identified predisposition to discharge in epileptiform bursts and suggest that they play an important role in the generation of seizure activity in the dentate gyrus. PMID- 16342373 TI - Exposure. Spit of HIV-positive man is considered 'deadly weapon'. PMID- 16342374 TI - Exposure. Man gets 21 years for exposing others to HIV. PMID- 16342376 TI - Medical marijuana. First government office opens to provide marijuana to patients. PMID- 16342375 TI - HIV-positive transplant patients live shorter lives. PMID- 16342377 TI - Correlation of central venous and arterial blood gas measurements in mechanically ventilated trauma patients. AB - HYPOTHESIS: Central venous blood gas (VBG) measurements of pH, PCO2, and base excess can be substituted for the same values obtained from an arterial blood gas (ABG) analysis in mechanically ventilated trauma patients, obviating the need for arterial puncture. DESIGN AND SETTING: Prospective comparison of 99 sets of VBGs and ABGs at a level 1 academic trauma center. PATIENTS: A consecutive sample of 25 trauma patients admitted to the intensive care unit who required mechanical ventilation and had both central venous and arterial catheters. MAIN OUTCOME MEASURES: Pearson correlations and Bland-Altman limits of agreement (LOAs) for pH, PCO2, and base excess values from each set of VBGs and ABGs. RESULTS: When VBG and ABG values were compared, pH had R = 0.92, P<.001, and 95% LOAs of -0.09 to 0.03; PCO2, R = 0.88, P<.001, and 95% LOAs of -2.2 to 10.9; and base excess, R = 0.96, P<.001, and 95% LOAs of -2.2 to 1.8. A receiver operating characteristic curve showed that a central venous PCO2 of 50 mm Hg had 100% sensitivity and 84% specificity for determining significant hypercarbia (arterial PCO2 > 50 mm Hg). CONCLUSIONS: Central venous and arterial PCO2, pH, and base excess values correlate well, but their LOAs represent clinically significant ranges that could affect management. Although VBGs cannot be substituted for ABGs in mechanically ventilated trauma patients during the initial phases of resuscitation, clinically reliable conclusions can be reached with VBG analysis. PMID- 16342379 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342378 TI - Evidence for the monophyletic evolution of benzylisoquinoline alkaloid biosynthesis in angiosperms. AB - Benzylisoquinoline alkaloids (BIAs) consist of more than 2500 diverse structures largely restricted to the order Ranunculales and the eumagnoliids. However, BIAs also occur in the Rutaceae, Lauraceae, Cornaceae and Nelumbonaceae, and sporadically throughout the order Piperales. Several of these alkaloids function in the defense of plants against herbivores and pathogens--thus the capacity for BIA biosynthesis is expected to play an important role in the reproductive fitness of certain plants. Biochemical and molecular phylogenetic approaches were used to investigate the evolution of BIA biosynthesis in basal angiosperms. The occurrence of (S)-norcoclaurine synthase (NCS; EC 4.2.1.78) activity in 90 diverse plant species was compared to the distribution of BIAs superimposed onto a molecular phylogeny. These results support the monophyletic origin of BIA biosynthesis prior to the emergence of the eudicots. Phylogenetic analysis of NCS, berberine bridge enzyme and several O-methyltransferases suggest a latent molecular fingerprint for BIA biosynthesis in angiosperms not known to accumulate such alkaloids. The limited occurrence of BIAs outside the Ranunculales and eumagnoliids suggests the requirement for a highly specialized, yet evolutionarily unstable cellular platform to accommodate or reactivate the pathway in divergent taxa. The molecular cloning and functional characterization of NCS from opium poppy (Papaver somniferum L.) is also reported. Pathogenesis- related (PR)10 and Bet v 1 major allergen proteins share homology with NCS, but recombinant polypeptides were devoid of NCS activity. PMID- 16342380 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342381 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342382 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342383 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342384 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342385 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342386 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342387 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342388 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342389 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342390 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342391 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342392 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342393 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342394 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342395 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342396 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342397 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342398 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342399 TI - Comments on Point:Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16342401 TI - Abstracts of the American Society of Hematology 47th annual meeting, December 10 13, 2005, Atlanta, Georgia, USA. PMID- 16342400 TI - Abstracts of the American Society of Hematology 47th annual meeting, December 10 13, 2005, Atlanta, Georgia, USA. PMID- 16342402 TI - Abstracts of the HUPO (Human Proteome Organisation) 4th Annual World Congress, August 29-September 1, 2005, Munich, Germany. PMID- 16342403 TI - [Ruggero Balli (1877-1958).Scientist, man, humanist, academician]. PMID- 16342404 TI - [Patient information. Ulcerous colitis]. PMID- 16342405 TI - Enhanced proliferation, attachment and osteopontin expression by porcine periodontal cells exposed to Emdogain. AB - Emdogain (EMD) is an enamel matrix derivative extracted from developing porcine teeth with demonstrated periodontal regenerative potential. EMD has been shown to influence a number of properties of periodontal ligament cells including proliferation, cell attachment and matrix synthesis. To date, the effect of EMD on the epithelial cell rests of Malassez (ERM) is unknown. In this study, periodontal ligament fibroblasts, ERM, alveolar bone cells and gingival fibroblasts were obtained from porcine periodontal ligament, alveolar bone and gingiva. This study investigated, in vitro, the effect of EMD at three concentrations on proliferation, cell attachment and expression of mRNA for two mineralised tissue-related proteins (osteopontin and bone sialoprotein). As for other periodontal cells, the ERM proliferative response was enhanced by EMD. Attachment assays revealed a highly significant increase for ERM and gingival fibroblasts after EMD treatment at all concentrations. This study has also shown that EMD stimulated expression of osteopontin mRNA by ERM and alveolar bone cells. The results from this study provide evidence that EMD enhanced cellular events related with proliferation, attachment and osteopontin mRNA expression by porcine periodontal cells, in a manner consistent with its role in periodontal regenerative therapy. PMID- 16342406 TI - Adoptive breastfeeding. PMID- 16342407 TI - Why do women stop breastfeeding? A closer look at 'not enough milk' among Israeli women in the Negev Region. AB - The aim of this study was to describe the breastfeeding experience of a sample of Israeli women. A random telephone survey of women, aged between 25 and 42, who had children under the age of 18 and were resident in the Negev Region of Israel (N=302) was conducted. Women had an average of three children each (range 1-11) and most breasted for less than three months. The most common reason given for not breastfeeding or breastfeeding less than three months was 'not enough milk' (90/205; 44%). A four-factor solution was extracted from a factor analysis of the reasons that women gave for stopping breastfeeding before three months or for not initiating breastfeeding. The four factors were: personal concerns (body image, tired, return to work); need help (wants husband to help, child unwell, didn't like breastfeeding), uncomfortable (nipple/breast pain, didn't like breastfeeding) and not confident (not enough milk). Women most frequently report that they stopped breastfeeding because they have insufficient milk, yet the lack of any consistent sociodemographic correlation indicates that this may be a universal way of expressing lack of confidence in breastfeeding. PMID- 16342408 TI - Turner's syndrome and breastfeeding. PMID- 16342409 TI - CK2 as a positive regulator of Wnt signalling and tumourigenesis. AB - CK2 is upregulated in rapidly dividing cells including most human tumours. Transgenic overexpression of CK2 in lymphoid or mammary lineages predisposes to transformation. Multiple signalling and oncogene pathways could be regulated by CK2 in this process. Our studies suggest that phosphorylation of critical oncogenes by CK2, as well as by other serine-threonine kinases, regulates their stability via susceptibility to the proteasomal degradation system. Beta-catenin is a transcriptional co-factor in the Wnt signalling pathway that is regulated in this fashion. Inactivating mutations in the adenomatosis polyposis coli (APC) gene, which encodes a carrier protein for beta-catenin, or stabilizing mutations in beta-catenin itself, frequently occur in human tumours. CK2 and the monomeric serine-threonine kinase GSK3 have opposing actions on beta-catenin: GSK-3 phosphorylation of the N-terminus of beta-catenin promotes degradation; while phosphorylation by CK2 in the armadillo repeat protein interaction domain protects it. Beta-catenin is overexpressed in mammary tumours occurring in mice transgenic for CK2 or a dominant negative form of GSK3, and also in mammary tumours arising following treatment with the environmental carcinogen DMBA. Experiments are underway to determine whether expression of both CK2 and kinase inactive GSK3 further accelerates tumorigenesis. Inhibitors of GSK3 under development for treatment of diabetes could promote tumours, while CK2 inhibitors should be useful agents for treatment of cancer. PMID- 16342410 TI - Downregulation of CK2 induces apoptosis in cancer cells--a potential approach to cancer therapy. AB - We have previously documented that naked antisense CK2alpha ODN can potently induce apoptosis in cancer cells in culture and in mouse xenograft human prostate cancer. The effects of the antisense CK2alpha are related to downregulation of CK2alpha message and rapid loss of the CK2 from the nuclear compartment. Here we demonstrate that downregulation of CK2 elicited by diverse methods leads to inhibition of cell growth and induction of apoptosis. The various approaches to downregulation of CK2 employed were transfection with kinase-inactive plasmid, use of CK2alpha siRNA, use of inhibitors of CK2 activity, and use of antisense CK2alpha ODN packaged in sub-50 nm nanocapsules made from tenascin. In all cases, the downregulation of CK2 is associated with loss in cell survival. We have also described preliminary observations on an approach to targeting CK2 in cancer cells. For this, sub-50 nm tenascin-based nanocapsules bearing the antisense CK2alpha ODN were employed to test that the antisense is delivered to the cancer cells in vivo. The results provide the first preliminary evidence that such an approach may be feasible for targeting CK2 in cancer cells. Together, our results suggest that CK2 is potentially a highly plausible target for cancer therapy. PMID- 16342411 TI - B23 is a downstream target of polyamine-modulated CK2. AB - Our previous studies have shown that the overexpression of ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine biosynthesis, increases the enzymatic activity of the polyamine-responsive enzyme casein kinase 2 (CK2). Because CK2 is known to preferentially associate with the nuclear matrix in response to other trophic stimuli, we investigated the effects of ODC overexpression on CK2 localisation and on the CK2-mediated phosphorylation of a known CK2 substrate, the nucleolar phosphoprotein B23. Immunofluorescence analysis of CK2 and B23 in primary keratinocytes revealed that ODC overexpression resulted in the colocalisation of CK2 with B23 at the nucleolar borders. ODC overexpression also increased CK2 kinase activity 2-fold at the nuclear matrix, a response which could be abrogated by treatment of K6/ODC transgenic keratinocytes with the ODC inhibitor alpha-difluoromethylornithine (DFMO). Levels of B23 protein were also elevated in ODC-overexpressing cells compared to normal cells or transgenic cells treated with DFMO. This increase in protein level was neither due to an increase in steady-state mRNA levels, nor was it due to increased stability of B23 protein. Phosphorylation of B23 was also increased in ODC overexpressing cells, and this increased phosphorylation could be blocked by treatment of the cells with the CK2 kinase inhibitors apigenin or 5,6-dichloro-1 beta-D-ribofuranosylbenzimidazole (DRB). These data suggest that B23 may be a downstream effector of polyamines via phosphorylation by the protein kinase CK2. PMID- 16342412 TI - A role for CK2alpha/beta in Xenopus early embryonic development. AB - CK2 is expressed widely in early embryonic development in several animal models, however its developmental role is unclear. One of the substrates of CK2 that is important in embryonic development is beta-catenin, the transcriptional co activator of the canonical Wnt signaling pathway. This pathway has been implicated in diverse aspects of embryonic development, including one of the earliest events in embryonic development, the establishment of the dorso-ventral embryonic axis. In Xenopus laevis, dorso-ventral axis formation is dependent upon stabilization of beta-catenin in the future dorsal side of the embryo. Since CK2 phosphorylation of beta-catenin stabilizes it, we hypothesized that CK2 might be critical to upregulation of beta-catenin in Xenopus embryos and to the process of axis establishment. Our results demonstrate that CK2 is required for dorsal axis formation and is for normal upregulation of Wnt signaling genes and targets. Thus, CK2 is a regulator of endogenous axis formation in vertebrates. PMID- 16342413 TI - Drosophila CK2 phosphorylates Deadpan, a member of the HES family of basic-helix loop-helix (bHLH) repressors. AB - In Drosophila, protein kinase CK2 regulates a diverse array of developmental processes. One of these is cell-fate specification (neurogenesis) wherein CK2 regulates basic-helix-loop-helix (bHLH) repressors encoded by the Enhancer of Split Complex (E(spl)C). Specifically, CK2 phosphorylates and activates repressor functions of E(spl)M8 during eye development. In this study we describe the interaction of CK2 with an E(spl)-related bHLH repressor, Deadpan (Dpn). Unlike E(spl)-repressors which are expressed in cells destined for a non-neural cell fate, Dpn is expressed in the neuronal cells and is thought to control the activity of proneural genes. Dpn also regulates sex-determination by repressing sxl, the primary gene involved in sex differentiation. We demonstrate that Dpn is weakly phosphorylated by monomeric CK2alpha, whereas it is robustly phosphorylated by the embryo-holoenzyme, suggesting a positive role for CK2beta. The weak phosphorylation by CK2alpha is markedly stimulated by the activator polylysine to levels comparable to those with the holoenzyme. In addition, pull down assays indicate a direct interaction between Dpn and CK2. This is the first demonstration that Dpn is a partner and target of CK2, and raises the possibility that its repressor functions might also be regulated by phosphorylation. PMID- 16342414 TI - A global view of CK2 function and regulation. AB - The wealth of biochemical, molecular, genetic, genomic, and bioinformatic resources available in S. cerevisiae make it an excellent system to explore the global role of CK2 in a model organism. Traditional biochemical and genetic studies have revealed that CK2 is required for cell viability, cell cycle progression, cell polarity, ion homeostasis, and other functions, and have identified a number of potential physiological substrates of the enzyme. Data mining of available bioinformatic resources indicates that (1) there are likely to be hundreds of CK2 targets in this organism, (2) the majority of predicted CK2 substrates are involved in various aspects of global gene expression, (3) CK2 is present in several nuclear protein complexes predicted to have a role in chromatin structure and remodeling, transcription, or RNA metabolism, and (4) CK2 is localized predominantly in the nucleus. These bioinformatic results suggest that the observed phenotypic consequences of CK2 depletion may lie downstream of primary defects in chromatin organization and/or global gene expression. Further progress in defining the physiological role of CK2 will almost certainly require a better understanding of the mechanism of regulation of the enzyme. Beginning with the crystal structure of the human CK2 holoenzyme, we present a molecular model of filamentous CK2 that is consistent with earlier proposals that filamentous CK2 represents an inactive form of the enzyme. The potential role of filamentous CK2 in regulation in vivo is discussed. PMID- 16342415 TI - Modulation of death receptor-mediated apoptosis by CK2. AB - Protein kinase CK2 has long been known to be involved in cell growth and proliferation. Recent work has also implicated its role in the suppression of apoptosis. We originally documented that removal of survival or growth stimuli resulted in rapid loss of CK2 from the nuclear matrix and chromatin which preceded induction of apoptosis. Further, we demonstrated that overexpression of CK2 in cells promotes suppression of drug-mediated apoptosis. In the present work, we have extended these observations to demonstrate that CK2 can influence apoptosis mediated via the death receptors. Overexpression of CK2 resulted in suppression of apoptosis mediated by TNF-alpha, TRAIL, and Fas-L in cells responsive to these ligands, whereas downregulation of CK2 resulted in augmentation of apoptosis mediated by these ligands. To our knowledge, this is the first report to show that receptor-mediated apoptosis can be modulated by changes in CK2 in prostate cancer cells. Based on our previous observations together with the evidence presented here, we propose that CK2 has an impact on the process of apoptosis mediated by diverse type of mechanisms thus playing a global role in regulation of apoptotic activity in cells. PMID- 16342416 TI - Exposure pathways and health effects associated with chemical and radiological toxicity of natural uranium: a review. AB - Natural uranium exposure derives from the mining, milling, and processing of uranium ore, as well as from ingestion of groundwater that is naturally contaminated with uranium. Ingestion and inhalation are the primary routes of entry into the body. Absorption of uranium from the lungs or digestive track is typically low but can vary depending on compound specific solubility. From the blood, two-thirds of the uranium is excreted in urine over the first 24 hours and up to 80% to 90% of uranium deposited in the bone leaves the body within 1.5 years. The primary health outcomes of concern documented with respect to uranium are renal, developmental, reproductive, diminished bone growth, and DNA damage. The reported health effects derive from experimental animal studies and human epidemiology. The Lowest Observed Adverse Effect Level (LOAEL) derived from animal studies is 50 microg/m3 for inhalation and 60 ug/kg body weight/day for ingestion. The current respiratory standard of the Occupational Safety and Health Administration (OSHA), 50 microg/m3, affords no margin of safety. Considering the safety factors for species and individual variation, the ingestion LOAEL corresponds to the daily consumption set by the World Health Organization Drinking Water Standard at 2 microg/L. Based on economic considerations, the United States Environmental Protection Agency maximum contaminant level is 30 microg/L. Further research is needed, with particular attention on the impact of uranium on indigenous populations, on routes of exposure in communities near uranium sites, on the combined exposures present at many uranium sites, on human developmental defects, and on health effects at or below established exposure standards. PMID- 16342417 TI - Evolution of function and color in GFP-like proteins. PMID- 16342418 TI - Reassembled GFP: detecting protein-protein interactions and protein expression patterns. PMID- 16342419 TI - The development of empathy-related responding. PMID- 16342420 TI - Sexual exposure to blood and behavioural risks among STI clinic patients in Cape Town, South Africa. AB - BACKGROUND: Exposure to blood during sexual intercourse is potentially important and yet understudied in HIV transmission. METHODS: The study included 415 men and 127 women receiving diagnostic and treatment services at a sexually transmitted infections (STI) clinic in Cape Town, South Africa, who completed anonymous sexual health behaviour surveys. RESULTS: More than 30% of both men and women reported engaging in sexual intercourse involving genital bleeding in the previous 3 months. Individuals who had engaged in sexual intercourse involving blood had significantly greater numbers of sex partners in the previous 3 months (OR = 1.6, 95%, CI = 1.1-2.3), engaged in higher rates of unprotected vaginal intercourse (OR = 1.2, 95%, CI = 1.1-1.2), were significantly more likely to have exchanged sex for money or materials (OR = 3.1, 95%, CI = 1.7-5.5) and were significantly more likely to have experienced condom breaks during intercourse (OR = 1.7, 95%, CI = 1.2-2.4). CONCLUSIONS: These findings suggest that exposure to blood during vaginal intercourse is prevalent among STI clinic patients in Cape Town and may be an important contributing factor to the rapid spread of HIV in South Africa. PMID- 16342421 TI - Condom use among brothel-based sex workers and clients in Bali, Indonesia. AB - BACKGROUND: The objective of this paper is to describe the situation among Indonesian sex workers and clients in Bali, Indonesia, through a series of studies conducted during the 1990s. METHODS: Five surveys of sex workers and clients were conducted from 1992 through to 1999. RESULTS: The proportion of female sex workers who have heard about AIDS and condom use has increased tremendously in this area. In 1992-93, 51% of women reported ever hearing of AIDS and this had increased to 97% by 1999 (P < 0.01). Reported condom use with clients increased from 19% in 1992-93 to 78% in 1999 (P < 0.01). However, condom use is still far from 100% and perceived susceptibility towards HIV infection among female sex workers and their male clients remains low. Ineffective preventive strategies such as medication use remain common. HIV infection was very low in this population, although the prevalence of other sexually transmitted infections such as gonorrhea and chlamydia was very high. CONCLUSIONS: Educational interventions have been conducted with the sex workers, but the main source of education for clients has been through the media. Future programs need to develop more focused efforts to reach clients. These interventions may include worksite interventions, mass media campaigns, youth programs or other models. PMID- 16342422 TI - Hepatitis A and B infection and vaccination in a cohort of homosexual men in Sydney. AB - OBJECTIVES: To determine the prevalence and incidence of hepatitis A (HAV) and B (HBV) infection and vaccination in HIV-negative homosexual men in Sydney, and associated risk factors. METHODS: An open prospective cohort study was conducted among a community-based sample of HIV-negative homosexual men in Sydney in 2001 02. Participants underwent a face-to-face interview, regarding demographics, sexual behavioural risk factors and sexually transmitted infections, and blood samples were collected. They were followed annually. RESULTS: Nine hundred and three men completed a baseline interview by the end of 2002. Among them, 68% were seropositive to hepatitis A. The seroprevalence of prior hepatitis B infection was 19%, and 53% had serological evidence of HBV vaccination. Younger men were much more likely to be seronegative, with 48% and 46% of <25-year-olds being seronegative to HAV and HBV respectively. In multivariate analysis HAV and HBV infection were associated with increasing age, greater number of lifetime sex partners and HBV infection was also associated with previous sexually transmitted infections. HAV vaccination was associated with increasing age, greater number of lifetime sex partners, overseas travel in the last year and self-reported anogenital warts. HBV vaccination was associated with higher occupational status, greater lifetime number of sex partners and previous sexually transmitted infections. CONCLUSION: Substantial proportions of gay community-attached young homosexual men are still at risk of HAV and HBV infection. This study points to a need for vaccination strategies which ensure high levels of hepatitis A and B immunity in young sexually active gay men. PMID- 16342423 TI - PKC alpha-dependent regulation of the IGF1 receptor in adult and embryonic rat cardiomyocytes. AB - In both, the adult rat ventricular cardiomyocytes and the embryonic rat heart cell line, H9c2, acute exposure to IGF1 resulted in activation of the IGF1 receptor's internal tyrosine kinase, and this was completely blocked by the PKC alpha inhibitor, Go6976. In addition, RNA interference using siRNA mediated gene silencing of PKC alpha-inhibited IGF1 receptor activity and blocked PKC alpha expression in H9c2 cells. Biochemical experiments demonstrate that PKC alpha is associated with the IGFlR (beta subunit) only after acute IGF1 exposure, and this may suggest that there is a direct interaction and possibly a PKC alpha phosphorylation site within the internal IGF1 receptor domain. The downstream effects of blocking PKC alpha activity by exposure to Go6976 include inhibition of IGF1-stimuated PI3 kinase activity and reduced IGF1-stimulated c-fos expression in the adult cardiomyocytes. Previously, the laboratory has reported that IGF1 activates PKC alpha in adult rat cardiomyocytes, and that PKC alpha activity is required for IGF1-dependent Erk/Erk2 activity and protein synthesis. Here, it is shown that IGF1-dependent protein synthesis is completely blocked by PD98059, indicating that the Raf-Mek-Erk cascade is required for IGF1's anabolic activity. Pretreatment with LY294002, a specific inhibitor of PI3 kinase, blocked IGF1-stimulated Erk1/Erk2 activity; therefore, PI3 kinase may also be required for IGF1-dependent protein synthesis. In H9c2 cells, coincubation with PMA lead to an increase in the rate of the IGF1 receptor activation, and this may further implicate a role for PKC in regulating the IGF1R. In conclusion, PKC alpha plays an essential role in the IGF1-signaling cascade, including the regulation of key signaling proteins involved in cell signaling and gene expression, and this may primarily be due to PKC alpha directly regulating the IGF1R. PMID- 16342425 TI - Swine in biomedical research: creating the building blocks of animal models. AB - The opportunities for utilizing swine biomedical models are immense, particularly in models that address lifestyle issues (nutrition, stress, alcohol, drugs of abuse, etc.). However, in order to fully capitalize upon the promise, there needs to be a more general recognition of these cofactors, such as nutrition, as key modulators of phenotype via genomic, epigenetic, and postgenomic mechanisms. Furthermore, increased interactions between nutrition scientists and clinical and fundamental researchers in other disciplines, including developmental biology, immunology, neuroscience, oncology, and cardiovascular and gastrointestinal physiology, are required. Closing discussions focused on the need for future conferences at more frequent intervals to support interactions between the various disciplines. This was especially critical because of the global distribution of investigators. PMID- 16342426 TI - Clinical tip: revision first metatarsophalangeal joint arthrodesis for sagittal plane malunion with an opening wedge osteotomy using a small fragment block plate. PMID- 16342424 TI - Angiotensin IV enhances phosphorylation of 4EBP1 by multiple signaling events in lung endothelial cells. AB - Angiotensin IV (Ang IV)-stimulated cell proliferation is regulated through activation of multiple signaling modules in lung endothelial cells (EC). Because eukaryotic intitiation factor 4E (eIF4E) binding protein 1 (4EBP1) plays a critical role in the RNA translation and the regulation of cell growth, we examined whether Ang IV modulates expression and/or phosphorylation of eIF4E and 4EBP1 as well as the role of multiple signaling events associated with 4EBP1 phosphorylation in EC. Ang IV stimulation increased phosphorylation but not expression of eIF4E and 4EBP1 proteins. Ang IV stimulation selectively phosphorylated Thr46 > Thr70 > Ser65 but not Thr37 residues in 4EBP1. Pretreatment of cells with PD-98059 and rapamycin, inhibitors of mitogen activated protein kinase (ERK1/2) and mammalian target for rapamycin (mTOR), respectively, partially blocked Ang IV-mediated phosphorylation of 4EBP1. In contrast, overexpression of p70 ribosomal S6 kinase (p70S6K) and protein kinase B (Akt) enhanced phosphorylation of 4EBP1 and eIF4E binding affinity to the cap region of mRNA. These results support critical roles of multiple signaling and phosphorylation of 4EBP1 by Ang IV in translation process and protein synthesis. PMID- 16342427 TI - Practical ethics. What would you do? AB - Your hospital receives a $1 million donation from a tobacco company executive. The money will be used to make a much-needed technology upgrade that will enable the hospital to report quality data, as well as improve the overall quality of care, a high priority in your community. The donation is from a private individual, but antismoking activists in the community are protesting the gift. The situation is generating a lot of bad publicity for the hospital and is beginning to overshadow recent positive news about improved care. Do you keep the money? PMID- 16342428 TI - Lesson learned: Texas hospitals braced for Hurricane Rita. PMID- 16342429 TI - Abuse of electroshock found in Turkish mental hospitals. PMID- 16342430 TI - Turkey's disabled. PMID- 16342431 TI - Bringing the two sides together. PMID- 16342432 TI - Writing that matters. PMID- 16342433 TI - Practical use of credit cards for ODs: does convenience outweigh the risks? PMID- 16342434 TI - Valuing the optometric practice. PMID- 16342435 TI - Atrial natriuretic peptide mRNA regulation by p38-MAPK in the perfused amphibian heart. PMID- 16342436 TI - I.B.M. to put genetic data of workers off limits. PMID- 16342437 TI - Guantanamo tour focuses on medical ethics. PMID- 16342438 TI - The problem with an almost-perfect genetic world. PMID- 16342439 TI - Hello, I'm your sister. Our father is donor 150. PMID- 16342440 TI - Dire wounds, a new face, and glimpse in a mirror. PMID- 16342441 TI - Push to loosen abortion laws in Latin America. PMID- 16342442 TI - South Korea's cloning crisis. PMID- 16342443 TI - A pioneering transplant, and now an ethical storm. PMID- 16342444 TI - She died the same way she lived: planning well in advance. PMID- 16342445 TI - Custody and abuse cases swirl around a troubled girl on life support. PMID- 16342446 TI - Ethical concerns on face transplants grow; American scientists raise medical and psychological issues. PMID- 16342447 TI - Found on the web, with DNA: a boy's father. PMID- 16342448 TI - Is this a solution? It may mollify some critics, but it's a stem cell shell game. PMID- 16342449 TI - Lab animal violations decried: activists urge NIH to sanction university for repeat offense. PMID- 16342450 TI - And now, acoustic ecology. PMID- 16342451 TI - What determines health? PMID- 16342452 TI - [Observation on the 78th meeting of the Japanese Society for Hansen's Disease]. PMID- 16342453 TI - Let's move on! PMID- 16342454 TI - A case of glimmering gloom. PMID- 16342455 TI - The mother of us all. PMID- 16342456 TI - Glory days. What price glory? PMID- 16342457 TI - Lessons from the ICU. PMID- 16342458 TI - Stinky people. Respecting your limitations & other people's predicaments. PMID- 16342459 TI - Full circle. PMID- 16342460 TI - "He keeps falling out" in the bathroom. PMID- 16342461 TI - Making hamburger of sacred cows: an interview with Dr. Bryan Bledsoe. Interview by Raphael M Barishansky. PMID- 16342462 TI - Chronology of a catastrophe. PMID- 16342463 TI - Five good reasons for better EMS documentation. PMID- 16342464 TI - How to take "no" for an answer. AB - Approximately 20% of all patients who are assessed by Council Bluffs Fire EMS providers refuse transport due to absence of any injury, minor injuries or illness that did not require medical attention. Using instruction sheets for this population has resulted in increased satisfaction with our EMS services. There are two important things to remember when educating personnel on the use of instruction sheets. First, the sheet does not replace the need for EMS providers to discuss their assessment findings with the patient. It is used to enhance the discussion and give the patient a written resource to refer to later. The patient hears the information from the provider, sees it on the information sheet and can review it and/or show it to a family member. Second, there should not be an increase in the number of refusals EMS personnel obtain. Providers should continue to encourage the patient to be transported to the hospital for further medical care and evaluation. Patient information sheets should never take the place of credible, well-written documentation on the signed refusal of care report. We have found that medics use the information sheets as resources during their documentation and communication with the patients. The medic documents in the refusal form that the information sheet was reviewed with the patient and the patient signs the refusal, acknowledging receipt of the form. Using standardized instruction sheets can improve both customer service and the image of prehospital care, as well as encourage providers to document and educate their patients who refuse transport. PMID- 16342465 TI - Postcards from the cutting edge. PMID- 16342466 TI - Uncompleted suicide attempts. PMID- 16342467 TI - Alternative remedies in the mainstream. PMID- 16342468 TI - Stress-control tools that start with your heart. PMID- 16342469 TI - Review of atmospheric water-soluble ionic species in Asia during 1998-2001. AB - Anthropogenic acid precursor emissions in Asia are increasing at a significant rate and this is expected to continue for the next several years. This paper comprises of studies that included sulfate, nitrate and ammonium around Asian countries (China, Korea, Japan and Taiwan) in recent years. The paper also provides the water-soluble ionic fine and coarse particulate concentrations in these Asian countries in recent years. Fine and coarse particulates in Taiwan (Kaohsiung and Talchung) and China (Nanjing and Shanghai) were found to have higher sulfate (average 18.68 and 15.75 microg/m3) and nitrate (average 12.77 and 7.71 microg/m3) concentrations than any other Asian country. The average sulfate concentrations for Japan and Korea were 7.31 and 5.84 microg/m3, respectively. The average nitrate concentrations for Japan and Korea were 6.73 and 4.65 microg/m3, respectively during the years 1998-2001. The different sample collection devices, analysis methods and pollutant concentrations are discussed in this study. The purpose of this study is to arrange the atmospheric water soluble ionic species (SO4(2-) and NO3) investigations in the Asian region. The data obtained here can also help to understand the sources, concentrations, phase distribution and health impact of atmospheric water-soluble ionic species (SO4(2 ) and NO3-) in other Asian regions. PMID- 16342470 TI - NDELA and nickel modulation of triazine disposition in skin. AB - Cutting fluids can become contaminated with metals (e.g., nickel, Ni) and nitrosamines (e.g., N-nitrosodiethanolamine, NDELA) and there is concern that these classes of contaminants can modulate dermal disposition and ultimately the toxicity of cutting fluid additives, such as irritant biocides (e.g., triazine). Biocides are added to these formulations to prevent bacterial degradation of commercial cutting fluids. The purpose of this study was to assess the dermal absorption and skin deposition of 14C-triazine when topically applied to porcine skin in an in vitro flow-through diffusion cell system as aqueous soluble oil (mineral oil, MO) or aqueous synthetic (polyethylene glycol, PEG) mixtures. 14C Triazine mixtures were formulated with NDELA and/or Ni, or with a combination of three additional cutting fluid additives; namely, 5% linear alkylbenzene sulfonate (LAS), 5% triethanolamine (TEA) and 5% sulfurized ricinoleic acid. Neither Ni nor NDELA was absorbed during these 8-h studies. However, 14C-triazine absorption ranged from 2.72 to 3.29% dose in MO and 2.29-2.88% dose in PEG with significantly greater triazine absorption in MO than PEG when all additives and contaminates were present. The difference between these two diluents was most pronounced when NDELA and/or Ni were present in cutting fluids. These contaminants also enhanced triazine deposition on the skin surface and skin tissues especially with PEG-based mixtures. In essence, the dermal disposition of irritant biocides could be dependent on whether the worker is exposed to a soluble oil or synthetic fluid when these contaminants are present. Workers should therefore not only be concerned about dermatotoxicity of these contaminants, but also the modulated dermal disposition of cutting fluid additives when these contaminants are present in cutting fluid formulations. PMID- 16342471 TI - Effect of water-soluble metal working fluid aerosols on respiratory system after 13 weeks of repeated inhalation exposure in F344 rats. AB - Three groups of male F344 rats were exposed to a water-soluble metal working fluid (MWF) aerosol at concentrations of 20, 60 or 180 mg/m3 for 6 h/day, five days a week, for 13 weeks in inhalation chambers. The aerosol particles were normally distributed and the mass median aerodynamic diameter was 1.56 microm. Despite the absence of clinical findings or significant changes in body weight during the 13-week exposure period, the numbers of white blood cells and lung weights were significantly higher at the end of the 13-week exposure period. Exposure to 20 mg/m3 of the MWF aerosol was found to have an effect on the respiratory system, including an accumulation of foamy macrophages in the bronchoalveolar lavage (BAL) fluid and thickening of the alveolar walls in the histopathology. The level of histamine and number of polymorphonuclear (PMN) cells were also higher in the BAL fluid from the rats exposed to 60 mg/m3 of the MWF aerosol, while the respiratory inflammation was most pronounced in the rats exposed to 180 mg/m3 of the MWF aerosol, including the accumulation of PMNs and foamy macrophages in the BAL cells, lung weight increase and thickening of the alveolar walls. Immunoglobulin IgG2a level was also lower in the sera from the rats exposed to 180 mg/m3 of the MWF aerosol. Therefore, even though no clinical symptoms were observed in the rats exposed to the high MWF concentration, respiratory inflammation was still induced by a relatively low concentration of the MWF, while the immune system was affected by the high MWF concentration. PMID- 16342472 TI - Inhaled vanadium pentoxide decrease gamma-tubulin of mouse testes at different exposure times. AB - Vanadium is an important environmental and industrial pollutant whose concentrations have increased in the last decades. Due to its status as reproductive toxicant and a microtubule damaging agent, the present study investigated by immunohistochemistry the effect of the inhalation of vanadium pentoxide on gamma-tubulin within somatic and testicular germ cells. Male mice inhaled vanadium pentoxide (V2O5) (0.02 M) 1 h/twice a week for 12 weeks. Our results demonstrated that vanadium accumulates in the testes starting with the initial inhalation (24 h), and this pattern remained until the last week of treatment. In general, vanadium was capable of significantly decreasing the percentage of gamma-tubulin in all analyzed testicular cells (Sertoli, Leydig and germ cells) starting with the first week of treatment. For all cell types studied, regression analysis revealed a negative and significant relationship between the percentage of immunopositive cells to gamma-tubulin and exposure time, showing a time dependent response in all cases. Our findings suggest that alterations on this protein might imply changes in microtubule-involved function such as cell division, which in the testes might lead to damage in the spermatogenesis, leading probably to infertility. PMID- 16342473 TI - Mobile phone-induced myocardial oxidative stress: protection by a novel antioxidant agent caffeic acid phenethyl ester. AB - Electromagnetic radiation (EMR) or radiofrequency fields of cellular mobile phones may affect biological systems by increasing free radicals, which appear mainly to enhance lipid peroxidation, and by changing the antioxidant defense systems of human tissues, thus leading to oxidative stress. Mobile phones are used in close proximity to the heart, therefore 900 MHz EMR emitting mobile phones may be absorbed by the heart. Caffeic acid phenethyl ester (CAPE), one of the major components of honeybee propolis, was recently found to be a potent free radical scavenger and antioxidant, and is used in folk medicine. The aim of this study was to examine 900 MHz mobile phone-induced oxidative stress that promotes production of reactive oxygen species (ROS) and the role of CAPE on myocardial tissue against possible oxidative damage in rats. Thirty rats were used in the study. Animals were randomly grouped as follows: sham-operated control group (N: 10) and experimental groups: (a) group II: 900 MHz EMR exposed group (N: 10); and (b) group III: 900 MHz EMR exposed+CAPE-treated group (N: 10). A 900 MHz EMR radiation was applied to groups II and III 30 min/day, for 10 days using an experimental exposure device. Malondialdehyde (MDA, an index of lipid peroxidation), and nitric oxide (NO, a marker of oxidative stress) were used as markers of oxidative stress-induced heart impairment. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities were studied to evaluate the changes of antioxidant status. In the EMR exposed group, while tissue MDA and NO levels increased, SOD, CAT and GSH-Px activities were reduced. CAPE treatment in group III reversed these effects. In this study, the increased levels of MDA and NO and the decreased levels of myocardial SOD, CAT and GSH-Px activities demonstrate the role of oxidative mechanisms in 900 MHz mobile phone induced heart tissue damage, and CAPE, via its free radical scavenging and antioxidant properties, ameliorates oxidative heart injury. These results show that CAPE exhibits a protective effect on mobile phone-induced and free radical mediated oxidative heart impairment in rats. PMID- 16342474 TI - Effect of 2,4-dichlorophenoxyacetic acid on the activities of some metabolic enzymes for generating pyridine nucleotide pool of cells from mouse liver. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D), which is a plant auxin analogue, is lethal to broad leaved weeds within days at high dosages and is considered as having low toxicity to mammals. Some studies have reported that exposure to this compound may cause damage to organs such as liver. The aim of this study was to investigate the effects of 2,4-D in mouse liver on chromosomes as well as hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) which are required for the generation of the pyridine nucleotide pool. The experiments were carried out with a 2,4-D group, an ethanol control for 2,4-D, and saline group for ethanol control group on three generations of mice. Only female parents were given 2,4-D during the gestation period, lactation period and for 33 days following the lactation period. In females of the first cross, 2,4-D caused a significant increase in the activity of LDH, and ethanol alone caused a significant increase in the activities of HK and LDH. In the male offspring of the first cross maternal, 2,4-D caused a significant increase in the activity of LDH, and ethanol alone caused a significant decrease in the activity of 6PGD. In the female offspring of the first cross maternal, ethanol caused a significant increase in the activities of G6PD and MDH. In the female offsprings of the third cross maternal, 2,4-D caused a significant increase in the activity of MDH. No gross morphological changes were observed in internal organs, such as liver, kidney and spleen of the affected animals. Also, a chromosomal study from bone marrow cells indicated no anomalies in chromosomal sets and structures. As a result, 2,4-D had an effect on the first cross maternal and their offsprings. The compound did not affect the parameters studied except MDH enzyme activity in the second and third generation of mice. PMID- 16342475 TI - Preventive effect of vitamin E on iron-induced oxidative damage in rabbit. AB - Although iron (Fe), plays an important role in different oxidative steps during the metabolism of the human body, it can cause free radical damage. Iron ions seem to play a major role in initiation and promotion reactions of intracellular lipid peroxidation. The aim of this study was to investigate if vitamin E has a protective effect on oxidative changes in erythrocytes induced by Fe treatment. Thirty male New Zealand white rabbits weighing 1400 +/- 50 g were used in the study. The animals were divided into three groups. The first group (n:10) was given 500 mg/kg iron-dextran through intraperitoneal (ip) injection. The second group was given 500 mg/kg iron-dextran+100 mg/kg vitamin E(ip). The third group constituted the control group and received a saline solution injection. The activities of erythrocyte antioxidant enzymes; Superoxide Dismutase (SOD), Glutatione peroxidase (GSH-Px), Catalase (CAT) and Malondialdehyde (MDA) level, an indicator of lipid peroxidation, were determined. Erythrocyte SOD, GSH-Px and CAT activities were decreased and MDA level was increased in iron-dextran treated animals compared to the control group (P < 0.05). The activities of the three antioxidant enzymes were increased and MDA level was decreased in iron-dextran and vitamin E treated group compared to the control group (P < 0.05). Our data indicate that lipid peroxidation occurs after iron overload in the blood. In the light of our findings, vitamin E administration can prevent the toxic oxidative effects induced by iron-dependent free radical damage in erythrocytes. PMID- 16342476 TI - A novel neural network for variational inequalities with linear and nonlinear constraints. AB - Variational inequality is a uniform approach for many important optimization and equilibrium problems. Based on the sufficient and necessary conditions of the solution, this paper presents a novel neural network model for solving variational inequalities with linear and nonlinear constraints. Three sufficient conditions are provided to ensure that the proposed network with an asymmetric mapping is stable in the sense of Lyapunov and converges to an exact solution of the original problem. Meanwhile, the proposed network with a gradient mapping is also proved to be stable in the sense of Lyapunov and to have a finite-time convergence under some mild condition by using a new energy function. Compared with the existing neural networks, the new model can be applied to solve some nonmonotone problems, has no adjustable parameter, and has lower complexity. Thus, the structure of the proposed network is very simple. Since the proposed network can be used to solve a broad class of optimization problems, it has great application potential. The validity and transient behavior of the proposed neural network are demonstrated by several numerical examples. PMID- 16342477 TI - Convergence analysis of a deterministic discrete time system of Oja's PCA learning algorithm. AB - The convergence of Oja's principal component analysis (PCA) learning algorithms is a difficult topic for direct study and analysis. Traditionally, the convergence of these algorithms is indirectly analyzed via certain deterministic continuous time (DCT) systems. Such a method will require the learning rate to converge to zero, which is not a reasonable requirement to impose in many practical applications. Recently, deterministic discrete time (DDT) systems have been proposed instead to indirectly interpret the dynamics of the learning algorithms. Unlike DCT systems, DDT systems allow learning rates to be constant (which can be a nonzero). This paper will provide some important results relating to the convergence of a DDT system of Oja's PCA learning algorithm. It has the following contributions: 1) A number of invariant sets are obtained, based on which we can show that any trajectory starting from a point in the invariant set will remain in the set forever. Thus, the nondivergence of the trajectories is guaranteed. 2) The convergence of the DDT system is analyzed rigorously. It is proven, in the paper, that almost all trajectories of the system starting from points in an invariant set will converge exponentially to the unit eigenvector associated with the largest eigenvalue of the correlation matrix. In addition, exponential convergence rate are obtained, providing useful guidelines for the selection of fast convergence learning rate. 3) Since the trajectories may diverge, the careful choice of initial vectors is an important issue. This paper suggests to use the domain of unit hyper sphere as initial vectors to guarantee convergence. 4) Simulation results will be furnished to illustrate the theoretical results achieved. PMID- 16342478 TI - Exponential stability of impulsive high-order Hopfield-type neural networks with time-varying delays. AB - This paper considers the problems of global exponential stability and exponential convergence rate for impulsive high-order Hopfield-type neural networks with time varying delays. By using the method of Lyapunov functions, some sufficient conditions for ensuring global exponential stability of these networks are derived, and the estimated exponential convergence rate is also obtained. As an illustration, an numerical example is worked out using the results obtained. PMID- 16342479 TI - Existence and global exponential stability of almost periodic solution for cellular neural networks with variable coefficients and time-varying delays. AB - In this paper, we study cellular neural networks with almost periodic variable coefficients and time-varying delays. By using the existence theorem of almost periodic solution for general functional differential equations, introducing many real parameters and applying the Lyapunov functional method and the technique of Young inequality, we obtain some sufficient conditions to ensure the existence, uniqueness, and global exponential stability of almost periodic solution. The results obtained in this paper are new, useful, and extend and improve the existing ones in previous literature. PMID- 16342480 TI - Extension neural network-type 2 and its applications. AB - A supervised learning pattern classifier, called the extension neural network (ENN), has been described in a recent paper. In this sequel, the unsupervised learning pattern clustering sibling called the extension neural network type 2 (ENN-2) is proposed. This new neural network uses an extension distance (ED) to measure the similarity between data and the cluster center. It does not require an initial guess of the cluster center coordinates, nor of the initial number of clusters. The clustering process is controlled by a distanced parameter and by a novel extension distance. It shows the same capability as human memory systems to keep stability and plasticity characteristics at the same time, and it can produce meaningful weights after learning. Moreover, the structure of the proposed ENN-2 is simpler and the learning time is shorter than traditional neural networks. Experimental results from five different examples, including three benchmark data sets and two practical applications, verify the effectiveness and applicability of the proposed work. PMID- 16342481 TI - PRSOM: a new visualization method by hybridizing multidimensional scaling and self-organizing map. AB - Self-organizing map (SOM) is an approach of nonlinear dimension reduction and can be used for visualization. It only preserves topological structures of input data on the projected output space. The interneuron distances of SOM are not preserved from input space into output space such that the visualization of SOM can be degraded. Visualization-induced SOM (ViSOM) has been proposed to overcome this problem. However, ViSOM is derived from heuristic and no cost function is assigned to it. In this paper, a probabilistic regularized SOM (PRSOM) is proposed to give a better visualization effect. It is associated with a cost function and gives a principled rule for weight-updating. The advantages of both multidimensional scaling (MDS) and SOM are incorporated in PRSOM. Like MDS, The interneuron distances of PRSOM in input space resemble those in output space, which are predefined before training. Instead of the hard assignment by ViSOM, the soft assignment by PRSOM can be further utilized to enhance the visualization effect. Experimental results demonstrate the effectiveness of the proposed PRSOM method compared with other dimension reduction methods. PMID- 16342482 TI - Finite-element neural networks for solving differential equations. AB - The solution of partial differential equations (PDE) arises in a wide variety of engineering problems. Solutions to most practical problems use numerical analysis techniques such as finite-element or finite-difference methods. The drawbacks of these approaches include computational costs associated with the modeling of complex geometries. This paper proposes a finite-element neural network (FENN) obtained by embedding a finite-element model in a neural network architecture that enables fast and accurate solution of the forward problem. Results of applying the FENN to several simple electromagnetic forward and inverse problems are presented. Initial results indicate that the FENN performance as a forward model is comparable to that of the conventional finite-element method (FEM). The FENN can also be used in an iterative approach to solve inverse problems associated with the PDE. Results showing the ability of the FENN to solve the inverse problem given the measured signal are also presented. The parallel nature of the FENN also makes it an attractive solution for parallel implementation in hardware and software. PMID- 16342483 TI - NDRAM: nonlinear dynamic recurrent associative memory for learning bipolar and nonbipolar correlated patterns. AB - This paper presents a new unsupervised attractor neural network, which, contrary to optimal linear associative memory models, is able to develop nonbipolar attractors as well as bipolar attractors. Moreover, the model is able to develop less spurious attractors and has a better recall performance under random noise than any other Hopfield type neural network. Those performances are obtained by a simple Hebbian/anti-Hebbian online learning rule that directly incorporates feedback from a specific nonlinear transmission rule. Several computer simulations show the model's distinguishing properties. PMID- 16342484 TI - The time dimension for scene analysis. AB - A fundamental issue in neural computation is the binding problem, which refers to how sensory elements in a scene organize into perceived objects, or percepts. The issue of binding is hotly debated in recent years in neuroscience and related communities. Much of the debate, however, gives little attention to computational considerations. This review intends to elucidate the computational issues that bear directly on the binding issue. The review starts with two problems considered by Rosenblatt to be the most challenging to the development of perceptron theory more than 40 years ago, and argues that the main challenge is the figure-ground separation problem, which is intrinsically related to the binding problem. The theme of the review is that the time dimension is essential for systematically attacking Rosenblatt's challenge. The temporal correlation theory as well as its special form--oscillatory correlation theory-is discussed as an adequate representation theory to address the binding problem. Recent advances in understanding oscillatory dynamics are reviewed, and these advances have overcome key computational obstacles for the development of the oscillatory correlation theory. We survey a variety of studies that address the scene analysis problem. The results of these studies have substantially advanced the capability of neural networks for figure-ground separation. A number of issues regarding oscillatory correlation are considered and clarified. Finally, the time dimension is argued to be necessary for versatile computing. PMID- 16342485 TI - The self-trapping attractor neural network--part II: properties of a sparsely connected model storing multiple memories. AB - In a previous paper, the self-trapping network (STN) was introduced as more biologically realistic than attractor neural networks (ANNs) based on the Ising model. This paper extends the previous analysis of a one-dimensional (1-D) STN storing a single memory to a model that stores multiple memories and that possesses generalized sparse connnectivity. The energy, Lyapunov function, and partition function derived for the 1-D model are generalized to the case of an attractor network with only near-neighbor synapses, coupled to a system that computes memory overlaps. Simulations reveal that 1) the STN dramatically reduces intra-ANN connectivity without severly affecting the size of basins of attraction, with fast self-trapping able to sustain attractors even in the absence of intra-ANN synapses; 2) the basins of attraction can be controlled by a single free parameter, providing natural attention-like effects; 3) the same parameter determines the memory capacity of the network, and the latter is much less dependent than a standard ANN on the noise level of the system; 4) the STN serves as a useful memory for some correlated memory patterns for which the standard ANN totally fails; 5) the STN can store a large number of sparse patterns; and 6) a Monte Carlo procedure, a competitive neural network, and binary neurons with thresholds can be used to induce self-trapping. PMID- 16342486 TI - Global exponential periodicity of a class of recurrent neural networks with oscillating parameters and time-varying delays. AB - In this paper, we present the analytical results on the global exponential periodicity of a class of recurrent neural networks with oscillating parameters and time-varying delays. Sufficient conditions are derived for ascertaining the existence, uniqueness and global exponential periodicity of the oscillatory solution of such recurrent neural networks by using the comparison principle and mixed monotone operator method. The periodicity results extend or improve existing stability results for the class of recurrent neural networks with and without time delays. PMID- 16342487 TI - Global exponential stability and global convergence in finite time of delayed neural networks with infinite gain. AB - This paper introduces a general class of neural networks with arbitrary constant delays in the neuron interconnections, and neuron activations belonging to the set of discontinuous monotone increasing and (possibly) unbounded functions. The discontinuities in the activations are an ideal model of the situation where the gain of the neuron amplifiers is very high and tends to infinity, while the delay accounts for the finite switching speed of the neuron amplifiers, or the finite signal propagation speed. It is known that the delay in combination with high gain nonlinearities is a particularly harmful source of potential instability. The goal of this paper is to single out a subclass of the considered discontinuous neural networks for which stability is instead insensitive to the presence of a delay. More precisely, conditions are given under which there is a unique equilibrium point of the neural network, which is globally exponentially stable for the states, with a known convergence rate. The conditions are easily testable and independent of the delay. Moreover, global convergence in finite time of the state and output is investigated. In doing so, new interesting dynamical phenomena are highlighted with respect to the case without delay, which make the study of convergence in finite time significantly more difficult. The obtained results extend previous work on global stability of delayed neural networks with Lipschitz continuous neuron activations, and neural networks with discontinuous neuron activations but without delays. PMID- 16342488 TI - Designing asymmetric Hopfield-type associative memory with higher order hamming stability. AB - The problem of optimal asymmetric Hopfield-type associative memory (HAM) design based on perceptron-type learning algorithms is considered. It is found that most of the existing methods considered the design problem as either 1) finding optimal hyperplanes according to normal distance from the prototype vectors to the hyperplane surface or 2) obtaining weight matrix W = [w(ij)] by solving a constraint optimization problem. In this paper, we show that since the state space of the HAM consists of only bipolar patterns, i.e., V = (v1, v2, . . ., vN)T E {-1, +1}N, the basins of attraction around each prototype (training) vector should be expanded by using Hamming distance measure. For this reason, in this paper, the design problem is considered from a different point of view. Our idea is to systematically increase the size of the training set according to the desired basin of attraction around each prototype vector. We name this concept the higher order Hamming stability and show that conventional minimum-overlap algorithm can be modified to incorporate this concept. Experimental results show that the recall capability as well as the number of spurious memories are all improved by using the proposed method. Moreover, it is well known that setting all self-connections wiiVi to zero has the effect of reducing the number of spurious memories in state space. From the experimental results, we find that the basin width around each prototype vector can be enlarged by allowing nonzero diagonal elements on learning of the weight matrix W. If the magnitude of w(ii) is small for all i, then the condition w(ii) = OVi can be relaxed without seriously affecting the number of spurious memories in the state space. Therefore, the method proposed in this paper can be used to increase the basin width around each prototype vector with the cost of slightly increasing the number of spurious memories in the state space. PMID- 16342489 TI - Design and analysis of a general recurrent neural network model for time-varying matrix inversion. AB - Following the idea of using first-order time derivatives, this paper presents a general recurrent neural network (RNN) model for online inversion of time-varying matrices. Different kinds of activation functions are investigated to guarantee the global exponential convergence of the neural model to the exact inverse of a given time-varying matrix. The robustness of the proposed neural model is also studied with respect to different activation functions and various implementation errors. Simulation results, including the application to kinematic control of redundant manipulators, substantiate the theoretical analysis and demonstrate the efficacy of the neural model on time-varying matrix inversion, especially when using a power-sigmoid activation function. PMID- 16342490 TI - Output feedback control of a class of discrete MIMO nonlinear systems with triangular form inputs. AB - In this paper, adaptive neural network (NN) control is investigated for a class of discrete-time multi-input-multi-output (MIMO) nonlinear systems with triangular form inputs. Each subsystem of the MIMO system is in strict feedback form. First, through two phases of coordinate transformation, the MIMO system is transformed into input-output representation with the triangular form input structure unchanged. By using high-order neural networks (HONNs) as the emulators of the desired controls, effective output feedback adaptive control is developed using backstepping. The closed-loop system is proved to be semiglobally uniformly ultimate bounded (SGUUB) by using Lyapunov method. The output tracking errors are guaranteed to converge into a compact set whose size is adjustable, and all the other signals in the closed-loop system are proved to be bounded. Simulation results show the effectiveness of the proposed control scheme. PMID- 16342491 TI - Speeding up the learning of robot kinematics through function decomposition. AB - The main drawback of using neural networks or other example-based learning procedures to approximate the inverse kinematics (IK) of robot arms is the high number of training samples (i.e., robot movements) required to attain an acceptable precision. We propose here a trick, valid for most industrial robots, that greatly reduces the number of movements needed to learn or relearn the IK to a given accuracy. This trick consists in expressing the IK as a composition of learnable functions, each having half the dimensionality of the original mapping. Off-line and on-line training schemes to learn these component functions are also proposed. Experimental results obtained by using nearest neighbors and parameterized self-organizing map, with and without the decomposition, show that the time savings granted by the proposed scheme grow polynomially with the precision required. PMID- 16342492 TI - Connectionist-based Dempster-Shafer evidential reasoning for data fusion. AB - Dempster-Shafer evidence theory (DSET) is a popular paradigm for dealing with uncertainty and imprecision. Its corresponding evidential reasoning framework is theoretically attractive. However, there are outstanding issues that hinder its use in real-life applications. Two prominent issues in this regard are 1) the issue of basic probability assignments (masses) and 2) the issue of dependence among information sources. This paper attempts to deal with these issues by utilizing neural networks in the context of pattern classification application. First, a multilayer perceptron neural network with the mean squared error as a cost function is implemented to calculate, for each information source, posteriori probabilities for all classes. Second, an evidence structure construction scheme is developed for transferring the estimated posteriori probabilities to a set of masses along with the corresponding focal elements, from a Bayesian decision point of view. Third, a network realization of the Dempster-Shafer evidential reasoning is designed and analyzed, and it is further extended to a DSET-based neural network, referred to as DSETNN, to manipulate the evidence structures. In order to tackle the issue of dependence between sources, DSETNN is tuned for optimal performance through a supervised learning process. To demonstrate the effectiveness of the proposed approach, we apply it to three benchmark pattern classification problems. Experiments reveal that the DSETNN out performs DSET and provide encouraging results in terms of classification accuracy and the speed of learning convergence. PMID- 16342493 TI - Neuron selection for RBF neural network classifier based on data structure preserving criterion. AB - The central problem in training a radial basis function neural network is the selection of hidden layer neurons. In this paper, we propose to select hidden layer neurons based on data structure preserving criterion. Data structure denotes relative location of samples in the high-dimensional space. By preserving the data structure of samples including those that are close to separation boundaries between different classes, the neuron subset selected retains the separation margin underlying the full set of hidden layer neurons. As a direct result, the network obtained tends to generalize well. PMID- 16342494 TI - SMO-based pruning methods for sparse least squares support vector machines. AB - Solutions of least squares support vector machines (LS-SVMs) are typically nonsparse. The sparseness is imposed by subsequently omitting data that introduce the smallest training errors and retraining the remaining data. Iterative retraining requires more intensive computations than training a single nonsparse LS-SVM. In this paper, we propose a new pruning algorithm for sparse LS-SVMs: the sequential minimal optimization (SMO) method is introduced into pruning process; in addition, instead of determining the pruning points by errors, we omit the data points that will introduce minimum changes to a dual objective function. This new criterion is computationally efficient. The effectiveness of the proposed method in terms of computational cost and classification accuracy is demonstrated by numerical experiments. PMID- 16342495 TI - Classifiability-based omnivariate decision trees. AB - Top-down induction of decision trees is a simple and powerful method of pattern classification. In a decision tree, each node partitions the available patterns into two or more sets. New nodes are created to handle each of the resulting partitions and the process continues. A node is considered terminal if it satisfies some stopping criteria (for example, purity, i.e., all patterns at the node are from a single class). Decision trees may be univariate, linear multivariate, or nonlinear multivariate depending on whether a single attribute, a linear function of all the attributes, or a nonlinear function of all the attributes is used for the partitioning at each node of the decision tree. Though nonlinear multivariate decision trees are the most powerful, they are more susceptible to the risks of overfitting. In this paper, we propose to perform model selection at each decision node to build omnivariate decision trees. The model selection is done using a novel classifiability measure that captures the possible sources of misclassification with relative ease and is able to accurately reflect the complexity of the subproblem at each node. The proposed approach is fast and does not suffer from as high a computational burden as that incurred by typical model selection algorithms. Empirical results over 26 data sets indicate that our approach is faster and achieves better classification accuracy compared to statistical model select algorithms. PMID- 16342496 TI - Posterior probability support vector machines for unbalanced data. AB - This paper proposes a complete framework of posterior probability support vector machines (PPSVMs) for weighted training samples using modified concepts of risks, linear separability, margin, and optimal hyperplane. Within this framework, a new optimization problem for unbalanced classification problems is formulated and a new concept of support vectors established. Furthermore, a soft PPSVM with an interpretable parameter v is obtained which is similar to the v-SVM developed by Scholkopf et al., and an empirical method for determining the posterior probability is proposed as a new approach to determine v. The main advantage of an PPSVM classifier lies in that fact that it is closer to the Bayes optimal without knowing the distributions. To validate the proposed method, two synthetic classification examples are used to illustrate the logical correctness of PPSVMs and their relationship to regular SVMs and Bayesian methods. Several other classification experiments are conducted to demonstrate that the performance of PPSVMs is better than regular SVMs in some cases. Compared with fuzzy support vector machines (FSVMs), the proposed PPSVM is a natural and an analytical extension of regular SVMs based on the statistical learning theory. PMID- 16342497 TI - Perceptual adaptive insensitivity for support vector machine image coding. AB - Support vector machine (SVM) learning has been recently proposed for image compression in the frequency domain using a constant epsilon-insensitivity zone by Robinson and Kecman. However, according to the statistical properties of natural images and the properties of human perception, a constant insensitivity makes sense in the spatial domain but it is certainly not a good option in a frequency domain. In fact, in their approach, they made a fixed low-pass assumption as the number of discrete cosine transform (DCT) coefficients to be used in the training was limited. This paper extends the work of Robinson and Kecman by proposing the use of adaptive insensitivity SVMs [2] for image coding using an appropriate distortion criterion [3], [4] based on a simple visual cortex model. Training the SVM by using an accurate perception model avoids any a priori assumption and improves the rate-distortion performance of the original approach. PMID- 16342498 TI - Using sensor habituation in mobile robots to reduce oscillatory movements in narrow corridors. AB - Habituation is a form of nonassociative learning observed in a variety of species of animals. Arguably, it is the simplest form of learning. Nonetheless, the ability to habituate to certain stimuli implies plastic neural systems and adaptive behaviors. This paper describes how computational models of habituation can be applied to real robots. In particular, we discuss the problem of the oscillatory movements observed when a Khepera robot navigates through narrow hallways using a biologically inspired neurocontroller. Results show that habituation to the proximity of the walls can lead to smoother navigation. Habituation to sensory stimulation to the sides of the robot does not interfere with the robot's ability to turn at dead ends and to avoid obstacles outside the hallway. This paper shows that simple biological mechanisms of learning can be adapted to achieve better performance in real mobile robots. PMID- 16342499 TI - A novel neural network for the synthesis of antennas and microwave devices. AB - A novel artificial neural network (SYNTHESIS-ANN) is presented, which has been designed for computationally intensive problems and applied to the optimization of antennas and microwave devices. The antenna example presented is optimized with respect to voltage standing-wave ratio, bandwidth, and frequency of operation. A simple microstrip transmission line problem is used to further describe the ANN effectiveness, in which microstrip line width is optimized with respect to line impedance. The ANNs exploit a unique number representation of input and output data in conjunction with a more standard neural network architecture. An ANN consisting of a heteroassociative memory provided a very efficient method of computing necessary geometrical values for the antenna when used in conjunction with a new randomization process. The number representation used provides significant insight into this new method of fault-tolerant computing. Further work is needed to evaluate the potential of this new paradigm. PMID- 16342500 TI - Neural-network-Based adaptive hybrid-reflectance model for 3-D surface reconstruction. AB - This paper proposes a novel neural-network-based adaptive hybrid-reflectance three-dimensional (3-D) surface reconstruction model. The neural network automatically combines the diffuse and specular components into a hybrid model. The proposed model considers the characteristics of each point and the variant albedo to prevent the reconstructed surface from being distorted. The neural network inputs are the pixel values of the two-dimensional images to be reconstructed. The normal vectors of the surface can then be obtained from the output of the neural network after supervised learning, where the illuminant direction does not have to be known in advance. Finally, the obtained normal vectors are applied to enforce integrability when reconstructing 3-D objects. Facial images and images of other general objects were used to test the proposed approach. The experimental results demonstrate that the proposed neural-network based adaptive hybrid-reflectance model can be successfully applied to objects generally, and perform 3-D surface reconstruction better than some existing approaches. PMID- 16342501 TI - Using adaline neural network for performance improvement of smart antennas in TDD wireless communications. AB - In time-division-duplex (TDD) mode wireless communications, downlink beamforming performance of a smart antenna system at the base station can be degraded due to variation of spatial signature vectors corresponding to mobile users especially in fast fading scenarios. To mitigate this, downlink beams must be controlled by properly adjusting their weight vectors in response to changing propagation dynamics. This can be achieved by modeling the spatial signature vectors in the uplink period and then predicting them to be used as beamforming weight vectors for the new mobile position in the downlink transmission period. We show that ADAptive LInear NEuron (ADALINE) network modeling based prediction of spatial signatures provides certain level of performance improvement compared to conventional beamforming method that employs spatial signature obtained in previous uplink interval. We compare the performance of ADALINE with autoregressive (AR) modeling based predictions under varying channel propagation (mobile speed, multipath angle spread, and number of multipaths), and filter order/delay conditions. ADALINE modeling outperforms AR modeling in terms of downlink SNR improvement and relative error improvement especially under high mobile speeds, i.e., V = 100 km/h. PMID- 16342502 TI - Spike timing dependent plasticity (STDP) can ameliorate process variations in neuromorphic VLSI. AB - A transient-detecting very large scale integration (VLSI) pixel is described, suitable for use in a visual-processing, depth-recovery algorithm based upon spike timing. A small array of pixels is coupled to an adaptive system, based upon spike timing dependent plasticity (STDP), that aims to reduce the effect of VLSI process variations on the algorithm's performance. Results from 0.35 microm CMOS temporal differentiating pixels and STDP circuits show that the system is capable of adapting to substantially reduce the effects of process variations without interrupting the algorithm's natural processes. The concept is generic to all spike timing driven processing algorithms in a VLSI. PMID- 16342503 TI - A 3-d surface reconstruction approach based on postnonlinear ICA model. AB - Photometric stereo technique deals with the reconstruction of three-dimensional (3-D) shape of an object by using several images of the same surface taken from the same viewpoint but under illuminations from different directions. In this paper, we propose a new photometric stereo scheme based on a new reflectance model and the postnonlinear (PNL) independent components analysis (ICA) method. The proposed nonlinear reflectance model consists of diffuse components and specular components for modeling the surface reflectance of a stereo object in an image. Unlike the previous approaches, these two components are not separated and processed individually in the proposed model. An unsupervised learning adaptation algorithm is developed to estimate the reflectance model based on image intensities. In this algorithm, the PNL ICA method is used to obtain the surface normal on each point of an image. Then, the 3-D surface model is reconstructed based on the estimated surface normal on each point of image by using the enforcing integrability method. Two experiments are performed to assess the performance of the proposed approach. We test our algorithm on synthetically generated images for the reconstruction of surface of objects and on a number of real images captured from the Yale Face Database B. These testing images contain variability due to illumination and varying albedo in each point of surface of human faces. All the experimental results are compared to those of the existing photometric stereo approaches tested on the same images. The results clearly indicate the superiority of the proposed nonlinear reflectance model over the conventional Lambertian model and the other linear hybrid reflectance model. PMID- 16342504 TI - Fast modular network implementation for support vector machines. AB - Support vector machines (SVMs) have been extensively used. However, it is known that SVMs face difficulty in solving large complex problems due to the intensive computation involved in their training algorithms, which are at least quadratic with respect to the number of training examples. This paper proposes a new, simple, and efficient network architecture which consists of several SVMs each trained on a small subregion of the whole data sampling space and the same number of simple neural quantizer modules which inhibit the outputs of all the remote SVMs and only allow a single local SVM to fire (produce actual output) at any time. In principle, this region-computing based modular network method can significantly reduce the learning time of SVM algorithms without sacrificing much generalization performance. The experiments on a few real large complex benchmark problems demonstrate that our method can be significantly faster than single SVMs without losing much generalization performance. PMID- 16342505 TI - Simultaneous perturbation learning rule for recurrent neural networks and its FPGA implementation. AB - Recurrent neural networks have interesting properties and can handle dynamic information processing unlike ordinary feedforward neural networks. However, they are generally difficult to use because there is no convenient learning scheme. In this paper, a recursive learning scheme for recurrent neural networks using the simultaneous perturbation method is described. The detailed procedure of the scheme for recurrent neural networks is explained. Unlike ordinary correlation learning, this method is applicable to analog learning and the learning of oscillatory solutions of recurrent neural networks. Moreover, as a typical example of recurrent neural networks, we consider the hardware implementation of Hopfield neural networks using a field-programmable gate array (FPGA). The details of the implementation are described. Two examples of a Hopfield neural network system for analog and oscillatory targets are shown. These results show that the learning scheme proposed here is feasible. PMID- 16342506 TI - A CMOS image sensor with programmable pixel-level analog processing. AB - A prototype of a 34 x 34 pixel image sensor, implementing real-time analog image processing, is presented. Edge detection, motion detection, image amplification, and dynamic-range boosting are executed at pixel level by means of a highly interconnected pixel architecture based on the absolute value of the difference among neighbor pixels. The analog operations are performed over a kernel of 3 x 3 pixels. The square pixel, consisting of 30 transistors, has a pitch of 35 microm with a fill-factor of 20%. The chip was fabricated in a 0.35 microm CMOS technology, and its power consumption is 6 mW with 3.3 V power supply. The device was fully characterized and achieves a dynamic range of 50 dB with a light power density of 150 nW/mm2 and a frame rate of 30 frame/s. The measured fixed pattern noise corresponds to 1.1% of the saturation level. The sensor's dynamic range can be extended up to 96 dB using the double-sampling technique. PMID- 16342507 TI - A new architecture for digital stochastic pulse-mode neurons based on the voting circuit. AB - This paper presents a new kind of architecture for artificial digital neurons based on the voting circuit, which may be considered an improved version of those presented in literature. Stochastic pulse modulation has been used, where the values of the neuron's inputs are coded in terms of bit probabilities. The resulting activation function closely resembles the logistic sigmoid, with a transition slope that can be selected at the architectural level with no additional hardware requirements. The proposed neuron architecture has been simulated in software. Simulation results confirm that the neuron features a sigmoid transfer characteristic similar to that of conventional voting circuits. The resource occupation of the neuron, as obtained from implementation on reconfigurable platforms, has been estimated to be significantly lower than previous implementations. The theoretical analysis of the neuron's behavior is also presented. PMID- 16342508 TI - Global exponential convergence of Cohen-Grossberg neural networks with time delays. AB - In this paper, we derive a general sufficient condition ensuring global exponential convergence of Cohen-Grossberg neural networks with time delays by constructing a novel Lyapunov functional and smartly estimating its derivative. The proposed condition is related to the convex combinations of the column-sum and the row-sum of the connection matrices and also relaxes the constraints on the network coefficients. Therefore, the proposed condition generalizes some previous results in the literature. PMID- 16342509 TI - Formulation and integration of learning differential equations on the stiefel manifold. AB - This letter aims at illustrating the relevance of numerical integration of learning differential equations on differential manifolds. In particular, the task of learning with orthonormality constraints is dealt with, which is naturally formulated as an optimization task with the compact Stiefel manifold as neural parameter space. Intrinsic properties of the derived learning algorithms, such as stability and constraints preservation, are illustrated through experiments on minor and independent component analysis (ICA). PMID- 16342510 TI - Deriving sufficient conditions for global asymptotic stability of delayed neural networks via nonsmooth analysis--II. AB - Following our recent approach of nonsmooth analysis, we report a new set of sufficient conditions and its implications for the global asymptotic stability of delayed cellular neural networks (DCNN). The new conditions not only unify a string of previous stability results, but also yield strict improvement over them by allowing the symmetric part of the feedback matrix positive definite, hence enlarging the application domain of DCNNs. Advantages of the new results over existing ones are illustrated with examples. We also compare our results with those related results obtained via LMI approach. PMID- 16342511 TI - Fast neural network ensemble learning via negative-correlation data correction. AB - This letter proposes a new negative correlation (NC) learning method that is both easy to implement and has the advantages that: 1) it requires much lesser communication overhead than the standard NC method and 2) it is applicable to ensembles of heterogenous networks. PMID- 16342512 TI - Exponential epsilon-regulation for multi-input nonlinear systems using neural networks. AB - This paper considers the problem of robust exponential epsilon-regulation for a class of multi-input nonlinear systems with uncertainties. The uncertainties appear not only in the feedback channel but also in the control channel. Under some mild assumptions, an adaptive neural network control scheme is developed such that all the signals of the closed-loop system are semiglobally uniformly ultimately bounded and, under the control scheme with initial data starting in some compact set, the states of the closed-loop system is guaranteed to exponentially converge to an arbitrarily specified epsilon-neighborhood about the origin. The important contributions of the present work are that a new exponential uniformly ultimately bounded performance is proposed and that the design parameters and initial condition set can be determined easily. The development generalizes and improves earlier results for the single-input case. PMID- 16342513 TI - A remark on "scalar equations for synchronous Boolean networks with biological applications" by C. Farrow, J. Heidel, J. Maloney, and J. Rogers. AB - The problem of finding all cycles in the exponentially growing state space of synchronous Boolean networks was studied in the paper by C. Farrow, J. Heidel, J. Maloney, and J. R. Scalar, "Equations for synchronous Boolean networks with biological applications," IEEE trans. Neural Networks, vol. 15, no. 2, pp. 348 354 Mar. 2004. No efficient algorithm was given to solve the problem. We show that even the determination of the number of fixed points (cycles of length 1) for monotone Boolean networks and the determination of the existence of fixed points for general Boolean networks are both strong NP-complete. PMID- 16342514 TI - [Haemostatic ointment efficacy in the treatment and prevention of epistaxis: a multi-centric study in 100 patients]. AB - OBJECTIVE: To evaluate the protective effects of an haemostatic ointment, at rhinosinusal mucosa level, on the development of bleeding in patients affected by epistaxis or with a positive clinical history for epistaxis in previous 7 days. PATIENTS AND METHODS: 100 patients affected by epistaxis or with a positive clinical history for epistaxis in previous 7 days. Patients underwent 15 days of treatment with an haemostatic ointment 2 times/day. RESULTS: The haemostatic ointment significantly reduced the percentage of patients affected by epistaxis and the number and severity of bleeding episodes (p < 0.001). 51% of patients experienced nasal obstruction during treatment, which persisted at the end of therapy only in 11% of them (p < 0.001). Nasal burning, nasal itching and rinorrhea involved less than 40% of patients in the first days of treatment and no more than 10% at the end of therapy (p < 0.001). CONCLUSIONS: The use of the haemostatic ointment in the treatment and in the prevention of not severe epistaxis acquire, in our opinion, a strong rationale. PMID- 16342515 TI - [Psoriasis and mud bath therapy: clinical-experimental study]. AB - OBJECTIVE: The psoriasis is chronic disease characterized from an acceleration of the kinetic of the cells of epidermis. To front of the empirical evidence of the benefits of the thermal therapy in the psoriasis, the experimentals-clinics studies is insufficient. The aim of research it has been that of quantify the benefits of the mud-bath therapy with mineral water in the psoriasis. PATIENT AND METHODS: The study has been channel on a champion of 30 subjects of which 19 of male sex and 11 of female sex with middle equal age to 56 years +/- 5.3 affected from psoriasis. The subjects of the examined champion have been divided to random in 2 groups: A and B. The group A has been treated with drugs used for psoriasis for 12 days; the B group has been treated, always for 12 consecutive days, with mud-bath therapy (FBT) with mineral water obtained from the mineral sources (chlorinate-sulphureous-bicarbonate) of the Spa of Stabia in Castellammare (NA). To the beginning and at the end of the advised treatments has been valued the prurient symptomatology and the PASI (Psoriasis and Severity Index). RESULTS: The data highlight an significant (P < 0.05) reduction is of the prurient symptomatology and of the PASI in both the groups considered. CONCLUSIONS: The results of this first step of investigations seems to highlight that the FBT treatment, to the same way of the drugs anti-psoriasis, results useful in the ameliorate the quality of life of these patients. PMID- 16342516 TI - Analysis of the chaotic component of the sinusal R-R intervals as a tool for detecting a silent cardiac dysautonomia in type 2 diabetes mellitus. AB - This study explores the heart rate (HR) variability (V) in order to detect whether the chaotic component of the sinusal R-R intervals (SRRI) can be interpreted as an early indicator of a silent cardiac neurovegetative dysautonomia in apparently uncomplicated Type 2 diabetic patients (DP). The SRRI were provided by the 24-h Holter ECG of 10 Type 2 DP (5 M and 5 F, mean age = 41 +/- 5 years). Control data were obtained by the 24-h Holter ECG of 10 clinically healthy subjects (CHS, 5 M and 5 F, mean age = 38 +/- 6 years). The chaotic component of HRV was investigated via the correlation dimension (CD) analysis (A) of the SRRI, performed per each hour of the ECG recording. The hourly-qualified series of SRRI, HR and CD index (I) were, in turn, analyzed via methods of conventional statistics and chronobiology, the latter ones for assessing the circadian rhythm (CR). The CDI CR was found to peak during the night in CHS, and to be unphysiologically rotated to the diurnal hours of the day in Type 2 DP. The diurnal inversion of the CDI CR in Type 2 DP suggests that the chaotic component of HRV shows an abnormal rhythnic pattern over the day-night period. Considering that the investigated Type 2 DP were lacking of documentable signs of cardiac neuropathy, it is hypothesized that the diurnal phase of shift CDI CR might be a potential indicator of a silent autonomic cardiac dysfunction in Type 2 DP. Such a hypothesis waits for further confirmations. PMID- 16342517 TI - [Nicotine addiction and current therapy of smoking cessation]. AB - Nicotine is defined as substance which provokes addiction because it creates both physiological and biochemical modifications in the nervous system stimulating the activity of dopaminergic neurons releasing dopamine in the areas of the brain that control pleasure. In this paper, after a short overview of neurobiological and cellular mechanisms involved in the pathway of nicotine addiction, the main therapies, used in order to provide support to smokers who decide to reduce their cigarette consumption or to quit smoking, are examined. These therapies can be enclosed in the following categories: nicotine replacement therapy (NRT), non nicotine pharmacological therapy (NNPT), psychological-behavioural therapies (PBT), alternative therapies (AT). In this work the advantages and disadvantages of various therapies are analysed, assessing the criteria found in literature. Results from randomised and controlled clinical studies which examine some of these therapies, alone or in association, also related to relapse time are reported. In conclusion, results of this analysis confirm that, as well as therapies and their treatment time, psychological support and personal motivation are indispensable for successful smoking cessation. PMID- 16342518 TI - [New CT and MR imaging techniques in non-traumatic neurological emergencies]. AB - New CT and MR imaging techniques used for non-traumatic neurologic emergencies (represented mostly by ischemic stroke) fulfil the exigency to know quickly and with high accuracy the presence of abnormalities in cerebral perfusion, with the final aim to practise immediately all the treatments needed to prevent the progression of the neurologic damage, by selecting those patients to undergo fibrinolysis, which is useless and not indicated in many occasions. The diagnosis of ischemia is only the first goal reached by these new diagostic tools, while it is nowdays possible and required to stratify the risk factors for the therapy and to accurate select those patients candidates to fibrolnilysis, in order to minimize the risck related to the inadequate treatment choice. PMID- 16342519 TI - [Hydrothorax in absence of ascites: an unusual complication of hepatic cirrhosis with portal hypertension]. AB - AIM: Authors, describing a clinical case of hepatic hydrothorax in absence of ascites, analyse the disease physiopathology and their therapeutic options. PATIENTS AND METHODS: Case report of a mixed aethiology (HCV and alcohol) hepatic cirrhosis, with pleural effusion, without ascites. CONCLUSIONS: Hepatic hydrothorax without ascites is an uncommon complication of cirrhosis with portal hypertension. Treatment could be pharmacological, with diuretics administration, or operating. A simple and cheap method is thoracentesis. If hydrothorax relapses, most effective method is transjugular intrahepatic portosystemic shunt. PMID- 16342520 TI - [Therapeutic advances in neoplastic hematology: target therapy anti-CD33]. AB - Antibodies capable to recognize antigen expressed on cancer cells represents the ideal approach for targeted anti neoplastic therapies. The CD33 antigen is present on 90% of acute myeloid leukemia blasts and is shared on normal hemopoietic cells only on the non stem dillerentiating fraction. Gemtuzumab Ozogamicin (GO) is an engineered humanized antibody anti-CD33 conjugated with a potent intercalating agent, named calicheamicin, which is release only at intracellular level (lower pH), following a selective binding to CD33-positive cells, thus representing a promising approach for target anti-leukemia therapy. GO was approved conditionally by the Federal Drug Administration in May 2000 as a single therapy for first recurrence of Acute Myeloid Leukemia (AML) in a subset of older patients. Since 2000, treatment trials and pilot studies have revealed potential expanded applications along with potential limitations. Phase II trials have confirmed the activity and the efficacy of GO as single agent in the treatment of relapsed AML. More recently, clinical trials on induction and post remission treatment of adult AML have shown efficacy of GO in combination chemotherapy. The strong and homogeneous CD33 expression in Acute Promyelocytic Leukemia (APL), have resulted in an effective treatment of this disease with GO used as salvage treatment, as well as innovative approach for molecular relapsed patients. However, the incidence of veno-occlusive disease, better defined as sinusoidal occlusive syndrome (SOS), must be taken into account as potential complication associated with the GO administration, especially in patients treated with ablative regimens. In conclusion, the extension of the approval in Italy to AML CD33+ in relapsed, regardless age limitation, along with the ongoing evaluation by the European EMEA, represent the basis for a large clinical application of GO in myeloid malignancies potentially extended to paediatric patients with AML and to ALL CD33+. PMID- 16342521 TI - What is your diagnosis? Type 1 Monteggia fracture. PMID- 16342522 TI - Dyslipoproteinemias and their clinical importance in several species of domestic animals. PMID- 16342523 TI - Evaluation of a behavioral assessment questionnaire for use in the characterization of behavioral problems of dogs relinquished to animal shelters. AB - OBJECTIVE: To evaluate a behavioral intake questionnaire in animal shelters for the presence of biased results and assess its use in the characterization of behavioral problems of dogs relinquished to shelters. DESIGN: Cross-sectional study. Animals-54 dogs being relinquished to a shelter and 784 dogs belonging to veterinary clients. Procedure-Owners who were relinquishing their dogs and agreed to complete the behavioral questionnaire were alternately assigned to 1 of 2 groups; participants were aware that information provided would be confidential or nonconfidential (i.e., likely used for adoption purposes). Data from confidential and nonconfidential information groups were compared, and the former were compared with data (collected via the questionnaire) regarding a population of client-owned dogs. RESULTS: Analyses revealed significant differences in 2 areas of reported problem behavior between the confidential and nonconfidential information groups: owner-directed aggression and stranger-directed fear. Compared with client-owned-group data, significantly more relinquished shelter dogs in the confidential information group were reported to have owner-directed aggression, stranger-directed aggression, dog-directed aggression or fear, stranger-directed fear, nonsocial fear, and separation-related behaviors. CONCLUSIONS AND CLINICAL RELEVANCE: Among persons relinquishing dogs to a shelter, those who believed questionnaire responses were confidential reported owner-directed aggression and fear of strangers in their pets more frequently than relinquishers who believed responses were nonconfidential. Confidentiality had no apparent effect on the reporting of other assessed behavioral problems. Results suggest that behavioral questionnaires may sometimes provide inaccurate information in a shelter setting, but the information may still be useful when evaluating behavior of relinquished dogs. PMID- 16342524 TI - Serum 17-alpha-hydroxyprogesterone and corticosterone concentrations in dogs with nonadrenal neoplasia and dogs with suspected hyperadrenocorticism. AB - OBJECTIVE: To assess serum 17-alpha-hydroxyprogesterone (17OHP) and corticosterone concentrations in dogs with nonadrenal neoplasia and dogs being screened for hyperadrenocorticism. DESIGN: Prospective study. ANIMALS: 16 clinically normal dogs, 35 dogs with nonadrenal neoplasia, and 127 dogs with suspected hyperadrenocorticism. PROCEDURE: ACTH stimulation tests were performed in all dogs. Baseline serum cortisol and corticosterone concentrations were measured in the healthy dogs; baseline serum cortisol concentration and ACTH stimulated cortisol, corticosterone, and 17OHP concentrations were measured in all dogs. Endogenous plasma ACTH concentration was also measured before administration of ACTH in dogs with neoplasia. RESULTS: In 35 dogs with neoplasia, 31.4% had high serum 17OHP concentration and 22.9% had high serum corticosterone concentration. Of the 127 dogs with suspected hyperadrenocorticism, 59 (46.5%) had high ACTH-stimulated cortisol concentrations; of those, 42 of 59 (71.2%) and 32 of 53 (60.4%) had high serum 17OHP and corticosterone concentrations, respectively. Of dogs with serum cortisol concentration within reference range after ACTH administration, 9 of 68 (13.2%) and 7 of 67 (10.4%) had high serum 17OHP and corticosterone concentrations, respectively. In the dogs with neoplasia and dogs suspected of having hyperadrenocorticism, post-ACTH serum hormone concentrations were significantly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: Serum concentrations of 17OHP or corticosterone after administration of ACTH may be high in dogs with nonadrenal neoplasia and no evidence of hyperadrenocorticism. Changes in serum 17OHP or corticosterone concentrations after administration of ACTH are proportionate with changes in cortisol concentration. PMID- 16342525 TI - Comparison between meloxicam and transdermally administered fentanyl for treatment of postoperative pain in dogs undergoing osteotomy of the tibia and fibula and placement of a uniplanar external distraction device. AB - OBJECTIVE: To compare the efficacy of meloxicam administered perioperatively with transdermal administration of fentanyl via a patch placed preoperatively in dogs undergoing orthopedic surgery. DESIGN: Prospective study. ANIMALS: 16 dogs. PROCEDURE: Unilateral or bilateral osteotomy of the tibia and fibula was surgically performed, and a uniplanar external distraction device was placed in each limb. Postoperative pain and lameness were assessed 24, 48, and 72 hours after administration of the first of 3 doses of meloxicam (0.2 mg/kg [0.09 mg/lb], IV, given preoperatively, followed by 0.1 mg/kg [0.045 mg/lb], IV, after 24 hours, and 0.1 mg/kg, PO, after 48 hours) or preoperative placement of a transdermal fentanyl patch (50 microg/h) left in place for 72 hours. RESULTS: No significant differences in total pain scores were detected between groups. Mean +/- SD lameness scores assessed at 24 and 72 hours were lower in dogs in the meloxicam group than dogs in the fentanyl group. Lameness scores decreased with time in a similar manner in both treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: Perioperative administration of meloxicam or preoperative placement of a transdermal fentanyl patch provided effective and similar postoperative analgesia in dogs undergoing orthopedic surgery. However, because of its anti inflammatory effects, treatment with meloxicam reduced the degree of lameness and resulted in rapid functional recovery of the limb. PMID- 16342526 TI - Analysis of the impact of trap-neuter-return programs on populations of feral cats. AB - OBJECTIVE: To evaluate 2 county trap-neuter-return (TNR) programs for feral cat population management via mathematical modeling. DESIGN: Theoretical population model. ANIMALS: Feral cats assessed from 1992 to 2003 in San Diego County, California (n = 14,452), and from 1998 to 2004 in Alachua County, Florida (11,822). PROCEDURE: Data were analyzed with a mathematical Ricker model to describe population dynamics of the feral cats and modifications to the dynamics that occurred as a result of the TNR programs. RESULTS: In both counties, results of analyses did not indicate a consistent reduction in per capita growth, the population multiplier, or the proportion of female cats that were pregnant. CONCLUSIONS AND CLINICAL RELEVANCE: Success of feral cat management programs that use TNR can be monitored with an easily collected set of data and statistical analyses facilitated by population modeling techniques. Results may be used to suggest possible future monitoring and modification of TNR programs, which could result in greater success controlling and reducing feral cat populations. PMID- 16342527 TI - Therapeutic percutaneous ultrasound-guided cholecystocentesis in three dogs with extrahepatic biliary obstruction and pancreatitis. AB - Three dogs were examined because of acute pancreatitis. In all 3, distension of the gallbladder was seen ultrasonographically, and extrahepatic biliary tract obstruction (EHBO) was diagnosed on the basis of ultrasonographic findings and serum biochemical abnormalities (i.e., high serum bilirubin and cholesterol concentrations and increased hepatic enzyme activities). In all 3 dogs, percutaneous ultrasound-guided cholecystocentesis (PUCC) was used to decompress the gallbladder, with cholecystocentesis performed multiple times in 1 dog. Serum bilirubin concentration was substantially decreased following the procedure in all 3 dogs. Two of the 3 dogs did not require surgery to resolve the obstruction. In the third dog, an exploratory laparotomy was performed because of concerns about development of abdominal effusion following the procedure. Bile staining of the mesenteric fat was seen during the laparotomy, but no defect in the gallbladder wall could be identified. In most dogs with EHBO secondary to pancreatitis, the obstruction resolves spontaneously as the acute pancreatitis improves so that surgery is not required. In those few dogs in which EHBO does not resolve or in which EHBO results in complications, therapeutic PUCC may be useful in relieving gallbladder distension. PMID- 16342528 TI - Characterization of the use of antiemetic agents in dogs with parvoviral enteritis treated at a veterinary teaching hospital: 77 cases (1997-2000). AB - OBJECTIVE: To characterize the use of antiemetic agents in dogs with canine parvovirus (CPV)-associated enteritis in a veterinary teaching hospital. DESIGN: Retrospective case series. ANIMALS: 77 dogs with CPV-associated enteritis. PROCEDURE: Medical records of 560 dogs with confirmed CPV-associated enteritis that were admitted to a veterinary teaching hospital were reviewed. Exclusion criteria included vaccination against CPV infection within the preceding 2 weeks, hospitalization for < 24 hours or removal from the hospital against advice, or an incomplete record. Signalment, duration of hospitalization, and daily antiemetic administrations were assessed; WBC counts and clinical findings were used to classify dogs as having systemic inflammatory response syndrome (SIRS). RESULTS: 77 dogs were included in the study; 55 (71%) received antiemetics (53 received metoclopramide at least once). Seventy-one dogs survived, and 6 dogs died (all 6 received antiemetics). Compared with dogs that did not receive antiemetics, duration of hospitalization was significantly longer for antiemetic-treated dogs. Daily values of rectal temperature and heart and respiratory rates did not predict administration of antiemetics or duration of hospitalization; however, compared with survivors, SIRS developed more frequently among nonsurvivors. Assessment of emetic events recorded hourly for 17 dogs indicated that antiemetic treatment did not control emesis. CONCLUSIONS AND CLINICAL RELEVANCE: Many dogs with CPV-associated enteritis had persistent vomiting despite antiemetic administration. The apparent difference in duration of hospitalization between antiemetic-treated dogs and other dogs may reflect a difference in disease severity between groups, although antiemetic-associated adverse events (e.g., signs of depression, hypotension, and immune modulation) may prolong hospitalization. PMID- 16342529 TI - Evaluation of survival rate and prognostic indicators for surgical treatment of left-to-right patent ductus arteriosus in dogs: 52 cases (1995-2003). AB - OBJECTIVE: To determine factors associated with long-term survival in dogs treated surgically for patent ductus arteriosus (PDA). DESIGN: Retrospective case series. Animals-52 dogs treated surgically for left-to-right shunting PDA. PROCEDURE: Data pertaining to age, breed, sex, body weight, clinical examination findings, type and duration of medical treatment, results of thoracic radiography and echocardiography, and surgical and postoperative complications were collected from records. Follow-up information was obtained from medical records or telephone interviews with owners or referring veterinarians. RESULTS: 22 dogs had mitral valve regurgitation. Mean weight and age were not significantly different between dogs with or without mitral valve regurgitation. Twenty-four (46.2%) dogs had clinical signs related to cardiac insufficiency. Left atrial dilatation was observed in 56.3% of dogs that were radiographed. Sonographic imaging was used to diagnose left atrial dilatation in 23 dogs and left ventricular dilatation in 25 dogs. The 1- and 2-year survival rates were 92% and 87%, respectively. Diagnosis of mitral valve regurgitation before surgery was not associated with the probability of survival. Age, weight, lethargy, preoperative treatment with angiotensin-converting enzyme inhibitors, and right atrial dilatation on radiographs at the time of surgery were negatively associated with probability of survival. CONCLUSIONS AND CLINICAL RELEVANCE: Surgical treatment of PDA was curative in young dogs without clinical signs of heart failure. Surgical correction of PDA should be recommended as early as possible after diagnosis, and mitral valve regurgitation is not a contraindication for surgery. PMID- 16342530 TI - Severe acute rhabdomyolysis associated with Streptococcus equi infection in four horses. AB - Four Quarter Horses (9 months to 7 years of age) with submandibular lymphadenopathy and firm muscles (palpation of which elicited signs of pain) were evaluated; in general, the horses had a stiff gait, and 3 horses became recumbent. Streptococcus equi was cultured from aspirates of lymph nodes or samples of purulent material collected from the auditory tube diverticula. Once the horses were recumbent, their condition deteriorated rapidly despite aggressive antimicrobial and antiinflammatory treatment, necessitating euthanasia within 24 to 48 hours. One horse did not become recumbent and recovered completely. Among the 4 horses, common clinicopathologic findings included neutrophilia, hyperfibrinogenemia, and high serum activities of creatine kinase and aspartate aminotransferase. Necropsies of the 3 euthanatized horses revealed large, pale areas most prominent in the semimembranosus, semitendinosus, sublumbar, and gluteal muscles that were characterized histologically by severe acute myonecrosis and macrophage infiltration of necrotic myofibers. Streptococcus equi was identified in sections of affected muscle by use of immunofluorescent stains for Lancefield group C carbohydrate and S. equi M protein. In the 4 horses of this report, acute severe rhabdomyolysis without clinical evidence of muscle atrophy or infarction was associated with S. equi infection; rhabdomyolysis was attributed to either an inflammatory cascade resembling streptococcal toxic shock or potentially direct toxic effects of S. equi within muscle tissue. PMID- 16342531 TI - Influence of NaCl on hydrogen production from glucose by anaerobic cultures. AB - In this study, batch experiments were conducted to investigate the inhibitory effects of NaCl on H2 production from glucose by anaerobic mixed cultures. Experimental results demonstrate that NaCl concentration had significant inhibitory effects on glucose degradation, cumulative H2 production, specific H2 production rate, H2 yield and the distribution of aqueous products. The specific H2 production rate had a negative correlation with NaCl concentrations. The NaCl concentrations, at which the specific H2 production rate and H2 yield were reduced by 50%, were estimated as 209 mM and 443 mM, respectively. PMID- 16342532 TI - Design strategy for a simultaneous nitrification/denitrification of a slaughterhouse wastewater in a Sequencing Batch Reactor: ASM2d modeling and verification. AB - Sequencing Batch Reactor (SBR) was used to treat slaughterhouse wastewater which contains average Chemical Oxygen Demand (COD) concentration of 5000 mg l(-1) and ammonium of 360 mgN l(-1). Nitrification/denitrification process was conducted for nitrogen removal. The influent wastewater as internal carbon source and sodium acetate as an external one was used for completing denitrification to achieve the simultaneous organic matter removal (95-96%) and nitrogen removal (95 97%). In addition, the dynamic SBR simulation model for biological nitrogen removal based on the Activated Sludge Model No. 2d (ASM2d) and GPS-X software is presented. The experimental study for the calibration and validation of the model was carried out using laboratory SBR. The study showed that the model provides a powerful tool to reduce the experimental expenditure and time to find the optimum strategy. PMID- 16342533 TI - Microbial community dynamics in manure composts based on 16S and 18S rDNA T-RFLP profiles. AB - Compost processing is assumed to be related to the microbial communities present. However, methods that will evaluate these relationships are not well understood. In this study, terminal restriction fragment length polymorphism (T-RFLP) analysis was used to evaluate the diversity of PCR-amplified bacterial 16S and fungal 18S rDNA communities from manure composts at different stages of composting (initial [day 0], thermophilic [day 24], and mature [day 104]). Results showed that the bacterial and fungal community profiles changed over the composting process, with bacterial communities showing a higher diversity compared with the fungal communities. During the thermophilic stage (day 24), the diversity of the bacterial communities increased, while the fungal communities decreased. As the compost reached maturity (day 104), a reverse pattern was observed between the diversity of bacterial and fungal communities. That is, the 18S rDNA T-RFLP-based diversity indices increased, while the 16S rDNA T-RFLP based diversity decreased. Differences in temperature profiles at different stages of composting impacted the chemical properties and the diversity of the microbial communities. The day 104 compost (mature) had lower water, organic matter and C contents and higher C and OM loss compared with the day 0 (initial) and day 24 (thermophilic) composts, which affected the diversity of the microbial communities. The results presented here demonstrated that distinctive community patterns from manure composts could be rapidly generated using T-RFLP analysis. The succession of peaks in combination of increasing and decreasing peak heights at different stage of composting indicates the high potential of T-RFLP technique to monitor the dynamics of microbial communities, and their variation qualitatively and quantitatively. PMID- 16342534 TI - Sustained and complete hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) degradation in zero-valent iron simulated barriers under different microbial conditions. AB - Flow-through columns packed with "aged" zero-valent iron (ZVI) between layers of soil and sand were constructed to mimic a one-dimensional permeable reactive iron barrier (PRB). The columns were continuously fed RDX (hexahydro-1,3,5-trinitro 1,3,5-triazine, ca. 18 mg l(-1)) for over one year. Two columns were bioaugmented with dissimilatory iron reducing bacteria (DIRB) Shewanella algae BrY or Geobacter metallireducens GS-15 to investigate their potential to enhance the reactivity of aged iron by reductive dissolution of passivating iron oxides or via production of biogenic reactive minerals. A third column was not bioaugmented to evaluate colonization by indigenous soil microorganisms. [14C]-RDX was completely removed in all columns at the start of the iron layer, and concentration profiles showed rapid and sustainable RDX removal over one year; however, a phylogenetic profile conducted after one year using DGGE analysis of recovered DNA did not detect S. algae BrY or G. metallireducens in their respective columns. Bacterial DNA was recovered from within the ZVI. Several unidentified 14C-labeled byproducts were present in the effluent of all columns. Dissolved 14C removal and the detection of dissolved inorganic 14C in these columns (but not in the sterile control) suggest microbial-mediated mineralization of RDX and sorption/precipitation of degradation products. Enhanced RDX mineralization in bioaugmented columns was temporary relative to the indigenously colonized column. However, shorter acclimation periods associated with bioaugmented PRBs may be desirable for rapid RDX mineralization, thereby preventing breakthrough of potentially undesirable byproducts. Overall, these results show that high RDX removal efficiency by ZVI-PRBs is achievable and sustainable and that the efficacy and start-up of ZVI-PRBs might be enhanced by bioaugmentation. PMID- 16342535 TI - Influence of bulking agents and microbial activator on thermophilic aerobic transformation of sewage sludge. AB - Bangkok, while improving the wastewater treatment in order to alleviate the river pollution, faces important amounts of sewage sludge. The sewage sludge contains organic matter, nitrogen and phosphorus available for plant growth. However, it may contain pathogenic microorganisms. To be used for agricultural purposes, these pathogens should be destroyed, which can be achieved with the thermophilic phase of composting. As the sewage sludge is dense and unable to compost alone (low C/N ratio), it should be mixed with an organic by-product. Two by-products available in large quantities in Thailand (wood chips and rice husk) have been tested for mixture with sewage sludge. As these products are not easy to decompose (presence of silica in rice husk and lignin/tannins in wood chips), the addition of a microbial activator for composting has been tested in controlled conditions (small quantities of organic mixtures, 55 degrees C, moisture maintained at 60-70% of water holding capacity). The monitoring of the decomposition has been made by measuring the carbon dioxide respiration, pH, organic matter and nitrogen contents and the evolution of enzymatic activities. When mixed with sewage sludge, wood chips and rice husk do not show significant differences concerning decomposition after 63 days. The use of an activator within the experimental conditions does not improve the decomposition of organic matter contained in the mixture of sewage sludge and rice husk or wood chips. PMID- 16342536 TI - Influence of the airflow rate on heat and mass transfers during sewage sludge and bulking agent composting. AB - The aim of this study was to characterize the gas flow during the composting, at a pilot scale, of a mixture of sludge and bulking agent, in order to model heat and mass transfers involved in the process. Thus, a closed 300-litre cylindrical pilot was fed with a mixture of wastewater treatment sludge and pine bark. Aeration was supplied from the bottom via an air blower and gases were collected at the top. Three experiments were led with constant gas flow rates and one with varying aeration rate. Temperatures within the pilot reactor were monitored all along the trials and their evolutions were discussed in term of heat transfers and parameters influencing the heat balance. Concurrently, Retention Time Distribution curves were obtained by injecting a pulse of methane in the entering airflow and by analysing the methane concentration in the exhaust gas, every two or three days during composting. The gas flow, within the composting medium, was characterized by a dispersion model, which is a deviation of the plug flow model. The dispersive effect of the flow was correlated to the evolution of the experimental temperature, and a convective dispersion model was used to describe the heat and mass transfers through the gas flow. These equations will be, in future work, coupled with heat production and mass degradation terms in order to model the global mass and heat balances of this composting process. Finally, axial dispersion coefficients of gases were determined and correlated with the airflow rate. PMID- 16342537 TI - Transport of Clostridium bifermentans DPH-1 through the laboratory column can be explained by two-region model for bioremediation. AB - Transport of Clostridium bifermentans DPH-1 was characterized from laboratory made column tests using fully saturated Toyoura sand. The conventional transport models were fitted to the column test results to investigate the applicability of prediction and assessment of bacterial transport in actual subsurface or ground water. Laboratory column tests confirmed that the transport characteristics of Clostridium bifermentans DPH-1 in activating tetrachloroethylene dechlorination could be described by mobile-immobile two-region model. The parameters of two region model i.e. peclet number, retardation factor, fraction rate of mobile water and stanton number were characterized by fitting results. These parameters were also justified by a verification experiment. Two-region model parameters suggested that bacterial injection into the ground at a large concentration is difficult for rehabilitation of widely dispersed contaminated ground water. PMID- 16342538 TI - Effects of hydrodynamic conditions on OH radical production at Ti/Pt anodes during electrochemical treatment. AB - The effects of hydrodynamic conditions on the production of hydroxyl radicals (*OH) during electrochemical treatment using a titanium coated by platinum anode and a stainless steel cathode are discussed in this paper. The sample used was ultra-pure water containing 200 mM of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). The amount of *OH for treatment was measured using electron spin resonance spectrometry coupled with DMPO spin trapping. Under constant hydrodynamic conditions, the production rate of the *OH spin adduct (DMPO-OH) increased with the terminal potential. This result was caused by the change in the electric current density at the anode, which was proportional to the rate of production of DMPO-OH. Increasing the linear velocity of water in an electrolytic cell promoted the production of DMPO-OH by two effects, namely, by the enhancement of ion transportation and the promotion of DMPO transfer from the bulk to the anode. The former effect emerged when the DMPO concentration near the anode was not insignificant in comparison with the DMPO concentration in the bulk, that is, when the overall rate-determining step was the electron transfer at the anode. The latter effect emerged when the DMPO concentration near the anode was much lower than the DMPO concentration in the bulk, that is, when the overall rate determining step was the diffusion of DMPO from the bulk to the anode. In addition, the latter effect was found to be proportional to the square root of the linear velocity of water in the electrolytic cell. PMID- 16342539 TI - [Drag-out in metal finishing industries]. AB - Currently, environmental regulations induce industrialists to implement source reduction techniques in order to comply with the prevention principle toward sustainable development. The project PIPSI (PIlotage Propre des Systemes Industriels/industrial system dean piloting) financing by Rhone-Alps Region is carried out with the aim to contribute to this objective. The study presented in this article concerns the pollution transfer in a metal finishing treatment line in order to minimise the environmental impact obtained notably with the pollution balance. Drag-out and draining phenomena have been particularly studied. Results obtained showed that a 10 seconds of draining reduced drag-out from 65 to 85% in terms of pieces design. Moreover, during the experiments, 5 drag-out levels were identified by medium values from 26 to 1700 ml m(-2). So that, either a piece can be associated to a level or knowing the piece drag-out level, it is possible to evaluate its medium drag-out value. Then the pollution balance will be obtained more easily. PMID- 16342540 TI - Selenate removal from sulfate containing aqueous solutions. AB - The performance of selenium remediation agents, mainly gamma alumina sorbents and zerovalent metals is greatly inhibited in the presence of large concentrations of sulfate found in selenium-rich agricultural drainage water. It is therefore imperative to remove sulfates from these contaminated waters, prior to their treatment for selenium removal. This paper presents data on enhanced selenium removal resulting from a two stage process involving the use of barium chloride in the first stage followed by a selenium remediation agent in the second. Barium chloride is known to stoichiometrically precipitate out sulfates. In addition to the sulfate removal from the contaminated solutions, a significant amount of selenate immobilization in the precipitated solids was achieved. When a loading of 15 g l(-1) of BaCl2, was employed, 100% selenate removal from an aqueous solution (containing 1 mg l(-1) selenate, 4 g l(-1) of sulfate and 2.5 g l(-1) of chloride) was achieved by the use of BaCl2, alone. Bimetallic NiFe particles, gamma alumina and activated carbon were employed to further remove selenium in the second stage. A solution containing 1 mg l(-1) and 4 g l(-1) of selenate and sulfate, respectively, when treated sequentially with 10 g l(-1) of BaCl2 and NiFe powder showed 100% selenium removal. PMID- 16342541 TI - [Nicotine patch--poor effect in ulcerative colitis. Not sufficiently efficient to justify therapeutic recommendation]. PMID- 16342542 TI - [The WHO global tobacco convention challenges Swedish physicians to "roll up the sleeves"]. PMID- 16342543 TI - [GHB, GBL and butanediol poisonings--a serious problem in Western Sweden]. AB - Acute poisoning with GHB (Gamma-hydroxybutyrate, Gamma-hydroxybutyric acid) has been an increasing medical and social problem during the last decade in Sweden, especially on the west coast. The number of poisonings decreased in the beginning of this millennium but has again increased during the last years. At the same time the number of seizures by the police has increased similarly as well as the number of drug-related deaths. During 2004 the number of GHB-abuse related deaths in western Sweden was seven, approximately the same figures as for heroin. Two other substances which are transformed to GHB in the human body, GBL (gamma butyrolactone) and 1,4-butanediol, have during the last years presented themselves among the same users as for GHB. Since GBL and butanediol are not classified as illicit drugs the possibilities for the police force to intervene and capture the drugs are severely restricted. Intoxication by GBL and butanediol has shown to be as dangerous as intoxication by GHB. Acute intoxications and abuse of these drugs is still a serious medical and social problem. A legal classification of GBL and butanediol as narcotics appears to be medically motivated. PMID- 16342544 TI - [Obstacles for recruiting organ donors in intensive care units can be overcomed. Identified obstacles are attitudes, ethics, resources and competence, as shown by an inquiry]. AB - The aim of this study was to identify obstacles for recruiting organ donors in the ICU. All specialists in neurosurgery and half of the specialists in anaesthesiology in Sweden were surveyed. Response rate was 67%. The following obstacles were identified: A neutral approach of the physicians when requesting organ donation of the families instead of a pro-donation approach; ethical problems concerning non-therapeutic ventilation and requesting donation of grieving families; varying competence in diagnosing brain infarction; lack of resources at the ICU. Suggestions for organizational and educational improvements are given. PMID- 16342545 TI - [Early outcome assessment following heart arrest is currently impossible. Clinical parameters are easy to use but afflicted with uncertainty]. AB - A critical review of a meta-analysis published in JAMA of early clinical neurological signs to detect "good" or "bad" outcome after cardiac arrest is presented. The review focuses on weaknesses of Glasgow Coma Scale, the Glasgow Pittsburgh Cerebral Performance Categories (CPC) and on individual differences in factors after arrest for study inclusion. It is suggested that the clinical assessments should be complemented with the assessment of neurochemical markers for brain damage. PMID- 16342546 TI - [Navigate correctly in PubMed. Strategies for retrieving clinically relevant information of sufficient quality]. PMID- 16342547 TI - [Tricuspid valve stenosis--an serious complication to Port-a-cath]. AB - Tricuspid valve stenosis and occlusion of superior vena cava are severe complications to Port-a-cath. In a child with SLE, symptoms started to develop about five to seven years after Port-a-cath insertion and cyclophosphamid injections. The patient developed hepatomegaly with abdominal and venous distension. Open heart surgery was necessary to remove the catheter. At operation it was found that the catheter was placed adjacent and through the tricuspid valve. The valve was severely stenosed with thrombus formation. The catheter and thrombus were removed, commissurotomy and bicuspidization of the valve and chordeal replacement performed to achieve an acceptable functional result. The superior vena cava was repaired with a pericardial patch. Retrospective analyses of the echocardiograms and chest x-rays show that the catheter was nearly related to the tricuspid valve and with the tip in the right ventricle. In such circumstances it is recommended with early withdrawal of the catheter, and in patients with immunological disease the indwelling time should be considered and limited. PMID- 16342548 TI - [Renewal is a general need]. PMID- 16342549 TI - [Pharaos' diseases surveyed with modern diagnostics]. PMID- 16342550 TI - [Directions by the authority for occupational environment will not allow unconditional substance abuse testing of school pupils]. PMID- 16342551 TI - [Adverse events are significantly more common than traffic accidents--and more expensive]. PMID- 16342554 TI - Passing the baton. PMID- 16342553 TI - I'm dreaming of a white paper... PMID- 16342552 TI - [Vitamin B12 and folate--an elucidation]. PMID- 16342555 TI - The power of partnerships. PMID- 16342556 TI - 100 not out. PMID- 16342557 TI - Ask the experts: person-centred care. PMID- 16342558 TI - 'Keeping well': a local approach to delivering health advice and information. PMID- 16342559 TI - Dementia services in rural areas: nursing implications. PMID- 16342560 TI - Understanding bowel problems in older people: Part 2. AB - Bowel problems can be devastating for patients and those who care for them. Accurate assessment is essential in determining the cause of symptoms and deciding treatment and management strategies. Treatment can be complex, but with care and patience improvement or cure is possible. Part 1 of this two-part article offered an overview of bowel problems in older people and an introduction to assessment. Part 2 discusses specific components of assessment, treatment and management. PMID- 16342561 TI - The wrong shoes: living with memory loss. PMID- 16342562 TI - Paid to be kind. PMID- 16342563 TI - It's OK to be old. PMID- 16342564 TI - Effect of arousal and retention delay on memory: a meta-analysis. AB - Two meta-analyses were conducted to examine the magnitude of the interaction between arousal and retention delay on various types of memory, e.g., verbal, visual, etc., as initially reported by Kleinsmith and Kaplan in 1963 and 1964. Results of the first meta-analysis (29 studies, N = 2,637) indicated that the interaction was robust (d = .779), with low arousal leading to better immediate memory than high arousal, whereas high arousal led to better delayed memory than low arousal. Incorporating additional studies, a subsequent meta-analysis (48 studies, N = 3,143) on the effect of arousal on memory, taking retention delay as a moderator, provided further support. At a 2-min. delay, low arousal led to better immediate memory than high arousal (d = -.459). This difference decreased at a 20-min. delay (d = .106), when directions of high and low arousal effects on memory appeared to reverse. At longer delays than 20 min., high arousal led to better delayed memory than low arousal (ds = .753, .219, and .472, for delays of 45-min., 1-day, and more than 1-day, respectively). PMID- 16342565 TI - Exploratory study of Machiavellianism and bases of social power in bankers. AB - This study focused on relationships between Machiavellianism in subordinates and perceived social power bases of supervisors in a sample of 168 bank officers in the western United States. Regression analyses showed no significant relationships between subordinates' Machiavellianism scores and four supervisory power bases subscales. The association between scores on Machiavellianism and coercive power reached p < .06. PMID- 16342566 TI - Associations of demographic information and philosophies of human nature. AB - A sample of 1117 students (53% women, 47% men; 50% 18-20 yr. old, 41% 21-23, 9% over 23 years) in classes on two university campuses completed a questionnaire comprised of the Philosophies of Human Nature Scale and demographic items. Factor analysis was conducted on the scale scores. Multiple linear regression was used to examine the strength of association between each factor and the 6 demographic traits. One-way analysis of variance comparing subjects' scale scores for each factor indicated significant association for demographic traits. The Scheffe test was used to identify differences among groups. Sex, age, employment status, year in school, and consistency in practicing faith orientation were associated with factors of the Philosophies of Human Nature Scale. PMID- 16342567 TI - Relations among measures of trait empathy, empathetic response, and willingness to get involved in customer-contact situations. AB - This study examined anticipated reactions to nonroutine occurrences in the context of emotionally laden customer-contact situations in retail stores. Correlations between measures of the dimensions of trait empathy, anticipated emotional responses to the situations, and self-rated willingness to be involved were examined. Anticipated Compassion mediated the relation of Empathetic Concern on Involvement Willingness in 3 of 4 imaginary compassion-evoking situations. No corresponding effect was observed for Anticipated Distress in the imaginary distress-evoking situations. PMID- 16342568 TI - A study of grouphate in a course on small group communication. AB - This study explored the relationship between grouphate and cohesion, consensus, relational satisfaction, affective learning, and cognitive learning. Participants were 83 undergraduate students enrolled in an introductory course on small group communication. Participants completed the Grouphate scale, the Classroom Cohesion scale, the Consensus scale, the Relational Satisfaction scale, three subscales of the Instructional Affect Assessment Instrument, and the Cognitive Learning Loss measure. Mean grouphate significantly increased during the semester, and negative correlations were found between scores for grouphate and cohesion (-.50), consensus (-.45), relational satisfaction (-.58), attitude toward the behaviors recommended in the course (-.23), the likelihood of developing an appreciation for the course content (-.33), and cognitive learning (-.32). Results may imply that students' grouphate is not associated with prosocial outcomes of the group work in this course. PMID- 16342569 TI - Sibship size, birth order, and personality. AB - In a sample of 273 American college students who were administered seven personality tests, only death obsession scores were consistently associated with sibship size and birth order (not optimism, pessimism, anxiety, a Taoist orientation, suicidal ideation, or obsessive-compulsive tendencies). PMID- 16342570 TI - Race of ratee and anonymity of rater: a study comparing students with practitioners as performance appraisers. AB - 123 students and 123 nonstudent supervisors viewed videotapes which displayed four supposed subordinate supervisors, two African Americans and two Caucasians, who individually described their respective performances during the past year. After being told either that the supposed subordinates would or that they would not have access to the performance rating, the subjects rated the performance of those subordinate supervisors. While anonymity of rater and race of rater had no evaluative effect on the performance ratings given by the nonstudent subjects, the student subjects gave higher ratings when they believed that their ratings would be made public. Also, the nonstudent subjects' ratings differed as a function of whether they worked closely with others of another race and as a function of the frequency with which they actually discussed performance evaluations with their own subordinates. PMID- 16342571 TI - Improvements in self-concept associated with reductions in negative mood in preadolescents enrolled in an after-school physical activity program. AB - Preadolescents enrolled in either a 3 times per week, 12-week exercise program (n = 50) or a no-exercise program control condition (n = 42) during their after school care, were assessed for indication of whether exercise-induced changes in self-concept and overall negative mood were negatively correlated. Significant improvements in ratings of self-concept and of negative mood were found over 12 wk. Inverse relationships were found between changes in physical self-concept with overall self-concept and negative mood (r = -.40 and -.36, respectively). Results supported the positive effect of exercise on the assessed psychological factors in preadolescents. Cognitive behavioral theory-based explanations of the findings were discussed. PMID- 16342572 TI - Stressful situations and coping strategies in relation to age. AB - Whether people cope differently with negative events at different ages was assessed by relation to age by type of situation perceived as potentially stressful and by type of coping strategy used, and also whether the kind of coping strategy used to reduce the potential stressor was related to type of stressor. To do this the factor structure of the Spanish version of the Ways of Coping Checklist of Lazarus and Folkman was examined in an Argentine sample of 153. The factor analysis of checklist scores identified five factors: Cognitive redefinition, Problem focus, Seeking social support, Wishful thinking, and Avoidance. For two groups, ages 20 to 24 and 40 to 45 years, analysis indicated a significant association of type of situation perceived as potentially stressful with age. The middle-age group (40-45 yr.) chose a way of coping in problem solution but the young adult group (20-24 yr.) seemed more often to elude problems. It appears that the type of stressor influences type of coping. PMID- 16342573 TI - Birth order and locus of control revisited: sex of siblings as a moderating factor. AB - Rotter's Locus of Control scale was administered to 64 first-born and 157 other undergraduate students, and data on the sex of participants' siblings were also obtained. The sex of participants' siblings was significantly correlated with locus of control among first-borns but not among other participants. PMID- 16342574 TI - A preliminary phenomenological study of being hypnotized and hypnotizing. AB - This paper presents phenomenological research conducted following Woodard's phenomenological and perceptual research methodology for understanding hypnotic experiencing. The research emphasizes examining the internal experiencings of individuals involved in hypnotic experiencing. Examples are presented of Individual Situated Structures and the General Structures from both a group of 8 participants who hypnotized their clients and another group of 17 individuals who volunteered to be hypnotized. The explicated themes identified in hypnotic experiencing (the hypnotic relationship, phenomenology of trance, use of imagination, problem with psychic energy, a gestalt of experiencing, and linear nonlinear experiencing) are discussed. The author discusses limitations of this study and suggestions for further work. PMID- 16342575 TI - Jehovah's Witnesses' child-rearing advice and potential psychologists' bias. AB - This study examined agreement with and potential bias toward child-rearing advice provided in publications of Jehovah's Witnesses. A sample of 750 APA member psychologists with Ph.D.s were randomly selected and assigned to one of three groups: (1) those informed about the source of the child-rearing advice, (2) those partially informed about the source of the advice, and (3) those uninformed about the source of the advice. Each group was provided identical materials except for identifying information about the specific source. Analyses show that psychologists report high agreement with the child-rearing advice provided by Jehovah's Witnesses through their publications, primarily The Watchtower and Awake! magazines. In general, however, psychologists expressed lower agreement with advice pertaining to early dating, premarital sex, and sports participation. Significantly lower participation was found in the group who were fully informed about the source of the advice. Implications of these findings are discussed and directions for research given. PMID- 16342576 TI - Graduate admissions criteria in psychology: an update. AB - Ratings of importance were reviewed for nine graduate admissions criteria for the 573 graduate degree programs listed in American Psychological Association's Graduate Study in Psychology (2003). Overall, the three criteria receiving the highest percentages of importance ratings were letters of recommendation, statement of goals and objectives, and grade point average. Importance ratings varied depending on the degree programs offered. Results are discussed in terms of the advice offered students interested in admission to graduate school and the importance of students tailoring their graduate admissions process to their specific degree program. PMID- 16342577 TI - Comorbidity and WISC-III profiles of Greek children with attention deficit hyperactivity disorder, learning disabilities, and language disorders. AB - The aim of the present study was to (a) examine the prevalence of ADHD and the comorbid difficulties in a sample of 114 children, 3.6 to 17.6 yr. of age (89 boys, 25 girls) referred to our Unit and (b) evaluate the discriminative ability of the WISC-III scores for children with Attention Deficit Hyperactivity Disorder (n = 22), Learning Disability (n = 50), and Language Disorder (n = 42). Analysis showed only 18% of 114 children had an Attention Deficit Hyperactivity Disorder diagnosis. Multivariate analysis of variance and stepwise discriminant function analysis were applied. Vocabulary and similarities were the best predictors for distinguishing between language disorders and the other two groups. Moreover, the Language Disorder group scored significantly lower on all the subtests while the Attention Deficit Hyperactivity Disorder and Learning Disability groups scored lower on coding and information, respectively. Children with Attention Deficit Hyperactivity Disorder and Learning Disability could not be accurately identified from the WISC-III test or their ACID profile. PMID- 16342578 TI - Job characteristics and burnout among aging professionals in information and communications technology. AB - The aim of this study was to examine the relationship between job characteristics and burnout, i.e., exhaustion, cynicism and lack of professional efficacy, in a sample of 115 (49- to 61-yr.-old) information and communications technology professionals. Questionnaire survey data were collected at two time points. In 1995 (Time 1), higher quantitative overload and lower job control were associated with higher exhaustion. Job control was negatively associated with lack of professional efficacy. In 2001 (Time 2), quantitative overload and information overload were positively associated with exhaustion, but with job control negatively. Use of new information was negatively associated with cynicism. In addition, job control and use of new information were negatively associated with lack of professional efficacy. Job characteristics at Time 1 were not significantly associated with burnout at Time 2 when job characteristics at Time 2 were controlled. PMID- 16342579 TI - Quantitative relation between conflict and response inhibition in the Flanker task. AB - The 2001 conflict monitoring hypothesis of Botvinick and colleagues posits that the amount of conflict raised by incongruent stimuli in a flanker task affects subsequent cognitive control, such as response inhibition. The present experiment yielded empirical evidence of the quantitative relation between conflict and response inhibition. Participants judged the direction of a target arrow flanked by distractor arrows presented above and below the target. The amount of conflict was manipulated by varying the distance between the target and the directional distractors. Analysis showed that response times were longer for incongruent trials than for congruent trials, and response times on incongruent trials were longer for the small distance than for the large distance conditions. In addition, the response times in congruent trials became longer as the amount of conflict in the preceding trial increased. These results are consistent with Botvinick, et al.'s hypothesis that the conflict-detection mechanism determines the amount of response inhibition depending on the amount of conflict. Responses on incongruent trials were faster and more accurate when the preceding trial was incongruent than when it was congruent, and the size of this response facilitation was not influenced by the amount of conflict. These results suggest that the conflict detection mechanism modulates the subsequent behaviors by two forms of control which are differently affected by the amount of conflict. PMID- 16342580 TI - An exploratory study of ethical values in nonprofit fundraising: survey of fundraising executives. AB - An initial study used the Ethical Position Quotient in an extension of this measure from for-profit organizations to nonprofit organizations. Responses from professional fundraising executives for nonprofit organizations were generally more idealistic than relativistic regarding ethical decision-making. Additionally, in a comparison of the executives' answers to those of M.B.A. students, the students were significantly more relativistic and less idealistic than the nonprofit executives. On Forsyth's Taxonomy of Ethical Perspectives, approximately 20% of the students were absolutists versus 66% of the fundraising executives. Conversely, 70% of the M.B.A. students were situationists compared to only a third of the executives. This disparity in responses between these groups provides a rich and interesting platform for research. PMID- 16342581 TI - Attribution and helping behavior: testing the attribution-affect-help judgment model in a Saudi sample. AB - A review of the literature on Weiner's attributional theory indicates that the theory has not been tested in Arabic cultures. The present study examined the reactions of Saudi men toward a friend with AIDS, using Weiner's attributional theory of social motivation. A sample of 298 Saudi men read two different vignettes about a friend with AIDS, in which the cause of AIDS was manipulated. Then the assignment of responsibility, reactions of anger and pity, and participants' willingness to provide help were measured. PMID- 16342582 TI - Emotional stability, positive and wishful thinking, and consequences of coping in a Spanish sample. AB - The relationship between scores on Emotional Stability and on two cognitive coping strategies-Positive Thinking and Wishful Thinking-and the Consequences of Coping scale were examined in a group of 99 Spanish undergraduates. Positive Thinking was associated with high Emotional Stability and positive consequences, whereas Wishful Thinking was associated with low Emotional Stability and negative consequences. PMID- 16342583 TI - Suicide among animals: clues from folklore that may prevent suicidal behaviour in human beings. AB - Knowing the most likely reasons for suicide might increase the chances to identify the early signs of suicide. Folkloric tales on suicide among animals are a possible source of such information, since people probably explain animal suicide using the same reasons they would apply to their kin. Modern naturalistic studies ave found little evidence of self-harming conduct among nonhuman species. Nevertheless, mythological accounts often report suicidal behaviour among animals. Claudius Aelian's De natura animalium, a classic in its genre, written in the 2nd century AD, reports 21 cases of suicide among animals. In Aelian's tales, the severing of social ties emerges as an important motive for suicide, together with incest and rage caused by adultery. Paying attention to the mechanisms leading to suicide described in ancient mythology may help us understand unusual and uncommon motives for suicide and the reasons people feel suicidal. PMID- 16342584 TI - Test-retest reliability of the relative work centrality measure. AB - The Meaning of Work International Research Team in 1987 developed the Relative Work Centrality measure, on which individuals divide 100 points among five major domains (work, leisure, community, religion, and family) in their lives, based on their relative centrality. The present study examined the test-retest reliability of this measure in a convenience sample of 96 Israeli full-time workers over a 4 wk. period. The sample consisted of 41.7% men and 58.3% women, of whom 48.9% were married. The participants had a mean age of 36.0 yr. (SD = 11.5) and a mean education of 14.9 yr. (SD = 2.5). The Spearman rank correlation coefficients for work, leisure, community, religion, and family at Time 1 and Time 2 were, respectively, .66, .76, .64, .76, and .82. These values indicate a fairly high test-retest reliability of the Relative Work Centrality measure and support the notion that this measure can also be applied to assess the relative centrality of each of the other four major life domains, in addition to work. PMID- 16342585 TI - Measuring direct and indirect aggression: behavior of is there a response bias? AB - Critics of self-report methods suggest that participants are likely to underreport their own negative behaviors given concerns about social desirability. The current study examined the problem of self-report bias by comparing individuals' estimates of their own and others' aggressive behavior. Undergraduate students (95 women, 50 men) completed the Richardson Conflict Response Questionnaire with regard to either their own behavior or that of another person of the same sex. As expected, participants reported significantly less aggression for themselves than for others. However, self and other ratings showed the same pattern of sex differences. These results suggest that, although individuals may underreport their own aggressive behaviors, this tendency may not affect relationships among variables. PMID- 16342586 TI - Are life satisfaction and self-esteem distinct constructs? A black South African perspective. AB - As part of a longitudinal project on Quality of Life, a study was undertaken to extend the applicability of the 5-item Satisfaction With Life Scale, developed in the USA, in South Africa. Data on basic sociodemographic characteristics, the scale, and the 10-item Rosenberg Self-esteem scale were available for 360 Black South Africans (151 men and 209 women), ages 21 to 83 years (M = 38.6 yr., SD = 10.3). Factor analysis applied to scale scores gave two factors, accounting for 71% of the variance. Factor I was loaded by 10 Self-esteem items and Factor II by four of the five Life Satisfaction items. Coefficient alpha was .77 for the Satisfaction With Life Scale and .97 for the Rosenberg Self-esteem Scale. Life Satisfaction was related to Self-esteem (r = .17, p < .01). It was concluded that Life Satisfaction and Self-esteem appear to be distinct, unitary constructs, but responses to Item 5 on the Satisfaction With Life Scale require cautious interpretation and may contribute to the weak r, although so may the collectivist culture of Black South Africans. PMID- 16342587 TI - Individualism and suicide. AB - In a sample of 19 nations, suicide rates were not associated with nations' estimates of work locus of control, job satisfaction, psychological well-being, physical well-being, or individualism. PMID- 16342588 TI - Psychosocial adaptation to spinal cord injury: a dimensional perspective. AB - From 313 individuals who sustained spinal cord injuries self-reported responses were examined on their psychosocial adaptation to disability and perceived quality of life. A multidimensional scaling analysis yielded a 2-dimensional structure of adaptation. The first dimension indicated adaptive versus nonadaptive reactions. The second dimension was suggestive of denial versus realization of the affect related to the medical impairment. PMID- 16342589 TI - The impact of the Samaritan centres on suicide in Scotland. AB - A study of Scottish suicide rates from 1958 to 1981 showed no evidence for a preventive effect from the growth in the number of Samaritan centres. PMID- 16342590 TI - Characteristics of true versus false allegations of sexual offences. AB - The goal of the study was to establish whether false allegations of sexual offences with an unknown perpetrator can be distinguished from accurate allegations. Case files of 27 true allegations of sexual offences with an unknown perpetrator were compared to those of 14 false allegations. The comparison was guided by a list of 43 criteria that are hypothesized to differentiate between true and false allegations of sexual assault. Analyses indicated the employed criteria differentiated true from false allegations to a certain extent. However, the discriminative strength of some criteria appeared to be stronger than that of others. Research is required to assess further the discriminative power of these criteria. PMID- 16342591 TI - Clinical utility and validation of the Couple's Communicative Evaluation Scale. AB - This study assessed the validity and clinical utility of a new test, the Couple's Communicative Evaluation Scale. With 24 couples from a variety of resources, e.g., churches, newspaper, and colleges, a discriminant analysis using the Dyadic Adjustment Scale, indicated that satisfied couples could be discriminated from issatisfied couples with 91-96% accuracy. Significant differences on the scale were found for means between 7 distressed and 16 nondistressed couples using the satisfaction/dissatisfaction cutoff score of 200 on the Dyadic Adjustment Scale and significant differences on the individual scales were found for means between 16 distressed and 31 nondistressed individuals using the satisfaction/dissatisfaction cutoff score of 100 on the Dyadic Adjustment Scale. Demographic variables, e.g., age, marriage length, were statistically significant. Scale scores were highly correlated with those on the Dyadic Adjustment Scale, indicating good validity. Using all 400 items, an alpha of .99 indicated good internal consistency for the verbal, nonverbal, and listening communication scores. PMID- 16342592 TI - Authoritarianism and sociopolitical attitudes in response to threats of terror. AB - The relationship between threat and authoritarianism in relation to support for democratic values and military aggression was examined. Participants completed Altemeyer's Right-Wing Authoritarianism Scale, read a scenario concerning 9/11/01 that was either threatening or nonthreatening, and completed the Democratic Values Scale and the Militarism Scale. Simple linear regressions indicate RWA scores contributed significant variation to Democratic Value Scale scores (43% of the variance) and Militarism Scale Scores (22% of the variance). There was a significant difference in the contribution of RWA scores between the threatening and nonthreatening conditions on the Democratic Values Scale (34% and 17%, respectively). These results suggest that in the presence of threat, there is increased activation of authoritarian responses. PMID- 16342593 TI - Exploratory behavior as a function of environmental novelty and complexity in male and female rats. AB - Laboratory rats show a positive response to low- and nonstressful novel events. The novel event may involve a number of aspects of the stimulus field. It is usually associated with a change in amount of environmental complexity. Most studies concerning novelty-related behavior involve the introduction of novel objects or the rearrangement of familiar objects. The present purpose was to measure exploratory behavior in response to environments of increased and decreased complexity. Both directions of environmental change are conditions of novelty. A two-way manipulation of increasing and decreasing the complexity of the environment was used. Rats of both sexes showed increased exploration to exposure to novelty, no matter which manipulation was applied; however, female and male rats behaved differently to the two types of novelty. Males responded more to novelty from the introduction of an unfamiliar object. The results indicate novel stimulation, whether of increasing or decreasing complexity, has reward properties. Perhaps the male-specific behavior directed toward unfamiliar objects may serve an adaptive function. PMID- 16342594 TI - Correlations among family environment, academic achievement, and academic attainment in a large sample of young Australian adults. AB - In a set of interrelated Australian and Spanish investigations in 2001-2004, relationships were examined among intellectual ability, family environment, and academic achievement. This follow-up study used longitudinal data from young Australian adults (4,397 men, 4,703 women; M age = 20.2 yr., SD = 0.4), to investigate relationships among earlier academic achievement, family influences, and academic attainment. From multiple regression and commonality analyses the findings indicated that earlier achievement and family influences were important as separate variables in explaining differences in later academic attainment (18.6% and 14.8%, respectively, for men; 14.5% and 11.5%, respectively, for women). In addition, earlier achievement and family influences reinforced each other to account for additional shared variance (9.1% for men, 6.3% for women) in young Australian adults' academic attainment. PMID- 16342595 TI - Dimensions of the Turkish version of the Psychological Empowerment Scale. AB - The purpose of the present study was to examine the dimensions of Spreitzer's Psychological Empowerment Scale for Turkish-speaking people. The scale was tested with a group of undergraduate students in Turkey (N = 214; M age = 22.6 yr., SD = 1.5, range = 19-27). Cronbach coefficient alpha was .84. By using maximum likelihood analysis with oblique rotation, three factors emerged and were labeled Influence, Meaning, and Competence. PMID- 16342596 TI - Smiling in photographs: childhood similarities between sexes become differences constant in adulthood. AB - Contemporary published photographs of 1639 children, 200 older teenagers, and 304 adults in North America were analyzed for the smiling pattern (full, partial, or none) exhibited by individuals by sex. For each of the three age groups and for both sexes, most individuals posed with a full smile. No statistically significant differences in smiling pattern were present between the sexes for photographs of children at preschool and grade-school age. By teenage years, and even more for the adults, there were statistically significant differences between the sexes with regard to a smiling facial expression. In those cases, more female than male subjects smiled fully, whereas more male than female subjects did not smile. Examination of photographs of well-known persons longitudinally through adulthood showed that individuals tended to be consistent in smiling pattern. There was no significant sex difference for this relative constancy of facial expression in posed photographs. PMID- 16342597 TI - A locus of control measure as a teaching demonstration. AB - 268 students (M age = 20.0 yr., SD = 4.4; 68% women), randomly chosen from psychology classes, completed Rotter's I-E scale and Pettijohn's 1992 locus of control scale for class demonstration. The two sets of scores were similar but examination of reliability and validity indicated the latter was best used as a demonstration. PMID- 16342598 TI - An examination of dimensions of Psychological Empowerment Scale for service employees. AB - In the present study the dimensions and factorial structure of the Psychological Empowerment Scale for a sample of 173 restaurant service employees from 20 restaurants in the USA were analyzed, using principal axis factor analysis. The analysis indicated a two-factor solution for the scale with Cronbach alpha of .91 and .89. These factors were labeled Attitude and Influence and accounted for approximately 63% of the common variance. This differed from the original four factor structure of Spreitzer and in two studies by Fulford and Enz in 1995 and by Hancer and George in 2003, a three-factor structure. Implications for research and recommendations are presented. PMID- 16342599 TI - Note on Meunier and Bodkins's (2005) "interpretation of MMPI-A Scale 5 with female patients". AB - Meunier and Bodkins discussed the unclear interpretation of Scale 5 of the MMPI A, investigated use of the scale with adolescent girls in a court-ordered residential treatment program and suggested that subclinical elevations on the scale may predict disruptive behavior in special samples. This note comments on statistics reported by Meunier and Bodkins and recommends additional analyses, including estimation of effect sizes for scales on which comparisons are statistically nonsignificant. PMID- 16342600 TI - Mental health issues in Arab society. PMID- 16342601 TI - Palestinian Psychiatric Association. PMID- 16342602 TI - Psychiatry and the Palestinian population. AB - Some key issues pertaining to Palestinian psychiatry are described. Bearing in mind the geographical location and history of the Palestinian population, the development of psychiatry needs to be seen in the context of the Arab world, on the one hand, and of Israel, on the other. In the Middle Ages, Arab culture and medicine were more developed than in Europe. Aspects of general and forensic psychiatry on the West Bank and Gaza are outlined. Issues pertaining to the death penalty, suicide and suicide bombers are also discussed. The biblically-described relationship between the Arab and Jewish peoples has its resonance today with regard to cohabitation and conflict. PMID- 16342603 TI - An update on mental health services in the West Bank. AB - The continuing development of the mental health services provided in the West Bank over the last fifty years is described. The hospital in Bethlehem was supplemented by the first community clinic in 1974, followed by clinics opening up in all 10 regions of the West Bank. The first Intifada brought actions from many international non-governmental organizations (NGOs). From 1993 the Palestine Authority's Ministry of Health has organized services, although the main addition initially was school counsellors from the Education Ministry. The second Intifada brought a further flurry of disorganized and well-intentioned interventions. A proposal for the reorganization of services from 2004 is in the course of implementation and should influence the training of personnel and structure of services. PMID- 16342604 TI - Child mental health problems in the Gaza Strip. AB - This study describes the mental health characteristics of 150 children of 6-13 years of age, who had been referred to different types of services in the Gaza Strip: a community mental health center, five primary health centers and a pediatric hospital. There was a high rate of somatising disorders among children referred to the mental health center (42%). Parent-reported rates of significant mental health problems were high for all groups, i.e., 70% in the mental health center group, 30% in the pediatric group, and 18% among children referred to primary health centers. PMID- 16342605 TI - Children living under a multi-traumatic environment: the Palestinian case. AB - This paper will focus on some of the most traumatic factors faced by the average Palestinian child during times of war. Unlike most research, which limits the Palestinian child's experience in war to military-related traumatic events, in this paper I will attempt to illustrate the Palestinian child's internal and external experiences, using empirical studies, my own clinical experience and an analytic understanding of historical and present events, in addition to a case study. This includes the different sources of stress and trauma that face Palestinian children and which ultimately create a multi-traumatic environment. I will also compare the situation of Palestinian children during the first and the second Intifadas. The analyses will include concepts such as trauma, witnessing, trans-generation, familial and social stress factors, and sense of loss of control. This paper could help clinicians as well as researchers to develop a more comprehensive conceptual model of understanding the emotional lives of Palestinian children. PMID- 16342607 TI - The impact of social values on the psychology of gender among Arab couples: a view from psychotherapy. AB - Three major psychosocial conditions have an effect upon the establishment of psychological problems within individuals and in the relationship of couples in the Arab family: (a) gender-dependent assessment of emotions, i.e., holding two different unequal yardsticks for two highly-genderized value systems; (b) imposing conflicting value demands on women; (c) glorifying and giving much respect to the notion of women's suffering in silence, a state described here as the "Mastoura" (tight lipped) woman, equivalent to the "learned helplessness" state. This article will also focus on the common forms of the manifestation of psychological or marital problems: sexual dysfunctions, somatic behavior and the usage of metaphoric language of psychosomatics. The article presents frequent complaints of couples who seek marriage therapy and the characteristics of each group. PMID- 16342606 TI - The psychological effects of Intifada Al Aqsa: acute stress disorder and distress in Palestinian-Israeli students. AB - The study assesses the effects of exposure to nationality-related and personal stressful events, threat appraisal and coping strategies on level of distress of Palestinian Israeli students. One hundred forty-eight Palestinian Israeli students filled out a battery of questionnaires that tapped their exposure to stressful life events, terrorism and political related violence, their primary and secondary appraisals, and coping strategies. Level of distress was evaluated by (1) acute stress disorder, and (2) psychiatric symptomatology. Results reveal relatively low exposure to terrorism-related traumatic events, yet considerable exposure (35.8%) to nationality-related stressful events during the last two years. Twenty-five percent of the students suffered from acute stress disorder, and their levels of psychiatric symptomatology exceeded norms for the general population. Primary appraisal processes and emotion-focused coping strategies made unique contribution to the respondents' level of (1) acute stress disorder and (2) psychiatric symptomatology. The implications of these findings are discussed. PMID- 16342608 TI - Mental health in Egypt. AB - The concepts and management of mental health in Egypt are presented from the Pharaonic era through the Islamic Renaissance until today. Papyri from the Pharaonic period show that Soma and Psyche were not differentiated and mental disorders were described as symptoms of the heart and uterus. Although theories of causation were of a mystical nature, mental disorders were treated on a somatic basis. In the Islamic era, mental patients were neither maltreated nor tortured as a consequence of the belief that they may be possessed by a good Moslem genie. In the 14th century mental disorders was one of the four departments in Cairo's Kalawoon Hospital, a precursor of the place of psychiatry in general hospitals that was accepted in Europe six centuries later. The mental health services in Egypt today are described, and transcultural studies carried out in Egypt of the prevalence and phenomenology of anxiety, schizophrenia, depression, suicide, conversion and obsessive compulsive disorders are reviewed. The psychiatric services for children are in their infancy. Since 1983 the common and semi-accepted use of hashish has been joined by abuse by heroin and other substances. PMID- 16342609 TI - Socio-political aspects of mental health practice with Arabs in the Israeli context. AB - Since the 1948 establishment of the Israeli state, an event described by Arab peoples as "Al-Nakbah" (catastrophe), the Arab minority in Israel has experienced oppression, trauma and social exclusion; they feel defeated, disempowered and poorer. There are huge gaps in quality of life between Arab and Jewish Israelis. Such social inequities, as well as other issues such as polygamy, have been identified as risk factors for psychological distress. This situation puts the Israeli Arab, like other post-colonial peoples, in an attitude of ambivalence towards modern mental health services. On the one hand, certain forms of intervention, particularly medicinal, may improve peoples' lives. On the other, mental health services, as part of the colonial process, continue to present limited cultural sensitivity towards Arab peoples. A cultural gap leading to mistrust is a given when a non-Arab mental health provider comes into contact with an Arab client. Religious beliefs, the importance of the family and the stigma attached to mental health problems have substantial influence on the Arab's perception and reaction toward mental health problems and their treatment. The expression of conflict and negative feelings are not well accepted within Arab culture. For this reason, mental illness is often denied and kept away from professional help or expressed as a physical illness. There is also a difficulty for a male being treated by a female and for the individual to ask for help outside his family or community. Arab Muslims also generally have a tendency to resign themselves to God's care and thus may neglect or deny symptoms. Another tendency is the preference for using traditional healers and folk medicine. Other problems in mental health work are the passive attitude of the patient and the degree of authority vested in the therapist. To facilitate bridging this cultural gap, the therapist's first task is that of educating him/her self about the religious, cultural and national background of the client. Cultural competence and self-reflection are key components to effective cultural practice. PMID- 16342610 TI - The importance of low serum levels of high-density lipoprotein cholesterol (HDL C) as a cardiovascular risk factor. AB - In order to discuss and establish a joint position on the treatment of low serum levels of high-density lipoprotein cholesterol (HDL-C), a group of experts involved in the care of people with dyslipidaemia and at risk of cardiovascular disease met in Miami, Florida, U.S., on 5th and 6th March 2005. The experts came from the Latin American countries Argentina, Brazil, Chile, Colombia, Ecuador, Guatemala, Mexico and Venezuela and had at least five years of experience in the care of patients with dyslipidaemia and low HDL-C. The main objective of the meeting was to discuss and propose a treatment for low serum HDL-C levels as a cardiovascular risk factor in patients and to create a group of useful recommendations in this regard, applicable to the daily clinical practice of physicians dealing with patients with dyslipidaemia and cardiovascular disease. This document describes the methodology developed to obtain these recommendations and presents the results of this academic meeting. PMID- 16342611 TI - Changes in serotonin metabolism in the rat brain on presentation of a habituated stimulus. AB - The levels of serotonin and the serotonin metabolite 5-hydroxyindoleacetic acid, monoamine oxidase activity, and the kinetic parameters of the oxidative deamination of serotonin were studied in various brain structures in rats after repeated presentation of a contextual stimulus. These experiments showed that presentation of the habituated stimulus was accompanied by increases in serotonin metabolism and the active transport of its metabolite in the amygdaloid complex, striatum, and midbrain, with no significant changes in the hippocampus or prefrontal cortex. Increases in monoamine oxidase activity in various brain areas resulted from different catalytic mechanisms: an increase in the rate of formation of the enzyme-substrate complex (a decrease in the Michaelis constant) in the amygdaloid complex, and faster conversion of the enzyme-substrate complex (increases in the maximum reaction rate) in the striatum and midbrain. It is concluded that activation of the presynaptic mechanism of serotonin transmission in the amygdaloid complex and striatum may be involved in the process of suppressing the biological significance of and attention to a repeatedly presented stimulus. PMID- 16342612 TI - Ischemic preconditioning of the rat brain as a method of endothelial protection from ischemic/repercussion injury. AB - The studies reported here addressed the endothelium-protecting action of local and remote ischemic preconditioning of the brain in rats. Cerebral ischemia lasting 30 min was reproduced by thermocoagulation of the vertebral arteries with simultaneous clamping of the carotid arteries, the procedure being followed by reperfusion via the carotid arteries for 120 min (controls). The early and late phases of ischemic preconditioning and remote preconditioning were reproduced. Brain blood flow was recorded using high-frequency Doppler ultrasonography. The early and late phases of local ischemic preconditioning and the late phase of remote ischemic preconditioning were found to have endothelium-protecting actions apparent as improvements in the recovery of brain blood flow in the post-ischemic period in preconditioned rats, with lower levels of endothelial desquamation and cerebral edema. Blockade of nitric oxide synthesis eliminated the protective effects of both phases of preconditioning. PMID- 16342613 TI - Acoustic aspects of the formation of speech in children in the third year of life. AB - This study covers the third year of life as part of a longitudinal investigation of the establishment of speech in Russian children performed on the basis of listener, phonetic, and instrumented acoustical analysis. The present report addresses the establishment of those additional acoustic and phonetic characteristics in children's speech which allow speech recognition. This is the first instrumented analysis in Russian children with statistical assessment of the dynamics of vowel formants in children's words, of the establishment of characteristics (stress, lack of stress), opposition (palatalization, lack of palatalization of consonants), and voice onset time for plosive consonants. The results showed that recognition of children's words by listeners with a high probability of success resulted from the formation of a system of acoustically stable properties in the children's speech which together provide informational adequacy for verbal communication. PMID- 16342614 TI - Hyperexcitability of neurons in field CAL1 evoked by transient episodes of hypoxia in hippocampal slices from rats of different ages. PMID- 16342615 TI - The effects of microinjection of the selective blocker of muscarinic M1 receptors pirenzepine into the neostriatum on the motor behavior of rats. AB - A discrimination conditioned active avoidance reflex (CAAR) model in a T maze was used in 18 rats to study the effects of bilateral microinjections of the selective muscarinic M1 receptor blocker pirenzepine into the neostriatum on the acquisition of the CAAR and behavior in an open field test. There was sharp degradation of learning of the CAAR and a significant improvement in motor activity both in the open field test and in the maze itself in rats given bilateral microinjections (pirenzepine, 0.004 mg) into the neostriatum as compared with intact controls. This suggests that changes in motor behavior (a sharp increase in locomotor activity) may be among the reasons for difficulty in learning the CAAR in rats after pirenzepine microinjections. Another reason for difficulty in learning the CAAR in these animals may be impairment of the perception of the conditioned signals (a flashing light) and poor differentiation. This is particularly indicated by the delay in the start chamber (double that seen in intact animals) on presentation of conditioned signals despite the high level of motor activity. These results and published data provide evidence for the complex nature of changes induced by blockade of muscarinic M1 receptors in the neostriatum. PMID- 16342616 TI - In vivo investigation of genome activity and synaptic plasticity of neurons in snails during learning. AB - The present report describes in vivo investigations of genome activity and its role in the mechanisms forming long-term synaptic plasticity in defensive behavior command neuron LPl1 during the acquisition of nociceptive sensitization by common snails. Transcription processes were recorded using SYTO 16, a specific fluorescent indicator of DNA activity, along with in vivo computer image analysis. Studies in control snails showed that application of nociceptive stimuli to the head led to biphasic changes in the bioelectrical responses of neurons to tactile and chemical stimulation--depression of responses in the short term stage (during the 1 h after training) and their facilitation during the long term stage of sensitization (more than 24 h). There were marked increases in fluorescence over the nucleus of the command neuron stained with SYTO 16 at 15-20 min from the start of training, this lasting 4-5 h. Acquisition of sensitization in the presence of the RNA synthesis inhibitor actinomycin D (20 microM) to the neuron led to the complete elimination of changes in fluorescence and synaptic facilitation in the responses of LPl1 to sensory stimulation in the long-term stage of sensitization but had no effect during the short-term stage of sensitization. Actinomycin D given 30 min after the end of acquisition of sensitization (1 h after the start) had no effect on the dynamics of fluorescence or synaptic facilitation. Thus, the acquisition of nociceptive sensitization is accompanied by a rapid (within 15-20 min) activation of the DNA of neuron LPl1 and subsequent (about 1 h) display of long-term synaptic facilitation. Induction of both processes was suppressed by the RNA synthesis inhibitor over a relatively short time period--1 h from the moment at which training started. PMID- 16342617 TI - Neuroeffector connections of giant multimodal neurons in the African snail Achatina fulica. AB - A new method of making preparations was used to analyse the neuroeffector connections of the paired giant neurons of the African snail Achatina fulica. These neurons were found to induce postsynaptic potentials in the muscles of the mantle, heart, the wall of the pulmonary cavity, and the muscular elements of the renal complex, the pericardium, the sexual apparatus, the walls of the cerebral arteries, the filaments of the columellar muscles, the wall of the abdomen, and the tentacle retractor muscles. Rhythmic neuron activity led to the development of marked facilitation and long-term potentiation of synaptic potentials. The possible significance of the multiple neuroeffector connections of giant neurons is discussed. PMID- 16342618 TI - The temperature sensitivity of the cholinergic responses of cortical neurons in the guinea pig brain. AB - Studies on slices of the parietal cortex of the guinea-pig brain showed that a change in temperature from 20 degrees C to 36 degrees C led to increases in responses to microintophoretic application of acetylcholine to individual nerve cells. The greatest changes occurred over two temperature ranges: 27-29 degrees C and 34-36 degrees C. Increases in the cholinergic response were accompanied by increases in the spontaneous activity of the neurons. PMID- 16342619 TI - Studies of damage to hippocampal neurons in inbred mouse lines in models of epilepsy using kainic acid and pilocarpine. AB - Identification of the mechanisms of damage to neurons is an important task in contemporary neuroscience and is of enormous importance in medicine. This report compares two models of neuron damage due to hyperexcitation induced by kainic acid and pilocarpine, using two lines of mice, C57BL/6J and FVB/NJ. Neuron damage was more marked in FVB mice, though lethality was greater in C57BL mice. The levels of convulsive activity were not significantly different. Kainic acid had greater tropism for the hippocampus than pilocarpine. Hsp-70 and Egr-1 expression was not significantly different in C57BL and FVB mice. Analysis of the isolated mitochondrial fraction showed that free radical production was different in these mouse lines; this may be one of the reasons for the differential resistance of neurons to hyperexcitation. PMID- 16342620 TI - Induction of NO synthase and glial acidic fibrillary protein in astrocytes in the temporal cortex of the rat with audiogenic epileptiform reactions. AB - The localizations of NADPH-diaphorase (NADPH-d), inducible NO synthase (iNOS), and glial acid fibrillary protein (GFAP) in astrocytes of the temporal cortex were studied in Krushinskii-Molodkina rats, which are genetically predisposed to audiogenic convulsive seizures. Convulsive reactions were induced in rats by three exposures to acoustic stimuli. Controls consisted of Wistar rats and Krushinskii-Molodkina rats not subjected to acoustic stimulation, these not developing convulsive reactions. The neocortex of animals with audiogenic convulsions consistently showed foci of brain tissue damage. Foci, of diameter 300-400 microm, were located in layers III-V and were groupings of NADPH-d positive astrocytes; these were seen in both hemispheres. Astrocytes in foci of damage expressed iNOS and had elevated GFAP levels. The numbers of GFAP immunopositive cells were increased by 25-37% in damage foci as compared with levels in controls and undamaged areas of the cortex. The induction of NO synthase and GFAP in astrocytes seen here indicates the involvement of glia in compensatory NO-dependent mechanisms formed in damage foci in response to audiogenic convulsive seizures. PMID- 16342621 TI - The cellular composition of the cerebral cortex of rat fetuses after fractionated low-dose irradiation. AB - Histological and morphometric studies were performed to address the characteristics of the morphogenesis of the sensorimotor cortex of the brain in rat fetuses subjected to fractionated gamma irradiation during the period from day 6 to day 18 of antenatal development at doses of 5, 25, 30, and 75 cGy. The results showed that fractionated irradiation at doses of 5-75 cGy had adverse effects on the processes of stem cell proliferation in the tissues of the developing cortex and also increased the intensity of cell destruction proportionally to the radiation dose. All cellular zones of the developing cortex showed increases in the absolute number of macroglial cells, which may be associated on the one hand with reactive increases in their production due to functional deficiencies and, on the other, with accelerated transformation of radial gliocytes into macroglial cells. Irradiation at doses of 5-75 cGy decreased the rate of migration of neuroblasts into the primary cortex, as shown by decreases in the numbers of cells in the neural differon in the neocortical rudiment and increases in their levels in the deep layers of the developing cortex. PMID- 16342622 TI - Immunocytochemical detection of astrocytes in brain slices in combination with Nissl staining. AB - The present study was performed to develop a simple and reliable method for the combined staining of specimens to allow the advantages of immunocytochemical detection of astrocytes and assessment of the functional state of neurons by the Nissl method to be assessed simultaneously. The protocol suggested for processing paraffin sections allows preservation of tissue structure at high quality and allows the selective identification of astrocytes with counterstaining of neurons by the Nissl method. The protocol can be used without modification for processing brain specimens from humans and various mammals--except mice and rabbits. PMID- 16342623 TI - The dynamics of the spatial synchronization of brain biopotentials in conditions of intense attention in the hypnotic state. AB - Twelve mentally healthy women aged 21-38 years were studied in the state of consciousness and hypnosis. The main study method was electroencephalography with assessment of the spatial synchronization of brain biopotentials (SSBP). Suggestion of high-intensity attention delivered to subjects in the hypnotic state was found to lead to significant reorganization of SSBP, with increases in SSBP between both occipital areas, the right temporal area, and other parts of the brain. The dynamics of brain SSBP in intense attention were opposite in the hypnotic and conscious states, which appears to result from the temporary exclusion in the hypnotic state of the functions of the frontal areas of the cortex responsible for conscious control and regulation of ongoing activity. PMID- 16342624 TI - The effects of systemic administration of selective antagonists of dopamine D1 and D2/D3 receptors on food-related and defensive (escape responses) conditioned paw-placing responses in cats. AB - Experiments were performed on cats to study the effects of systemic administration of antagonists of dopaminergic transmission on food-related and defensive (an escape response) operant conditioned reflexes acquired on the basis of the innate response of placing the forepaw on a support. Selective blockade of D1 receptors with SCH23390 (0.005-0.1 mg/kg) completely and selective blockade of D2/D3 receptors with raclopride (0.1-0.25 mg/kg) partially suppressed both reflexes. At these doses, both blockers had stronger actions on the defensive conditioned escape reflex than the food-related reflex: SCH23390 had significantly stronger inhibitory effects on both reflexes than raclopride. PMID- 16342625 TI - Dopaminergic modulation of spike activity in sensorimotor cortex neurons in a conditioned reflex. AB - Changes in the spike conditioned reflex reactions of sensorimotor cortex neurons during microiontophoretic application of agonists and antagonists of glutamatergic and GABAergic transmission and their modulation by dopamine were studied. A paradoxical reaction is described, consisting of facilitation of spike responses evoked by antagonists of ionotropic glutamate transmission, which was blocked by GABA. This is evidence for the active involvement of inhibition in organizing the excitatory responses of neurons in the conditioned reflex. Application of the metabotropic glutamate transmission antagonist MCPG was accompanied by sharp suppression of the baseline and evoked activity of cortical neurons, along with increases in the latency of spike responses and conditioned reflex movement. Dopamine was found to reverse the effect of blocking metabotropic glutamate receptors and to normalize neuron activity, which is evidence for the stabilizing role of dopamine in the functioning of neocortical neurons. PMID- 16342626 TI - Organization of frontohippocampal neuronal networks in cats in different types of directed behavior. AB - Food-related operant conditioned reflexes to light were developed in four cats on the basis of the "active choice" of reinforcement quality: short-latency pedal presses were reinforced with a mixture of meat and bread, while long-latency presses were reinforced with meat. Animals showed differences in their behavioral strategies: two preferred long-latency pedal presses (animals with "self control"), while the other two preferred short-latency pedal presses ("impulsive" animals). At the second stage of the study, animals of both groups were retrained to a short-delay (1 sec) conditioned operant food-related reflex in response to light with meat reinforcement. Chronically implanted Nichrome semimicroelectrodes were used to record multicellular activity in the frontal cortex and hippocampus (field CA3). The interaction of neighboring neurons within the frontal cortex and hippocampus (local neural networks) and neurons of the frontal cortex and hippocampus (distributed frontohippocampal and hippocampofrontal neural networks) were assessed by statistical cross-correlation analysis of spike trains with an analysis epoch of 100 msec. The frontal and frontohippocampal neural networks had different modes of functional organization in the simplified task for the animals of the two groups. However, intergroup differences in local networks of the hippocampus persisted in conditions of the simplified task lacking the requirement for the animals to select the quality of the reinforcement, indicating the likely genetic determinacy of these networks. PMID- 16342627 TI - ADN educators: shine your light! PMID- 16342628 TI - Keeping current: on persistently questioning our teaching practice. PMID- 16342629 TI - HIPAA and nursing education: how to teach in a paranoid health care environment. AB - Nursing faculty are affected by the paranoia that exists today as health care institutions individually interpret the Privacy Rule contained within the Health Insurance Portability and Accountability Act of 1996 (HIPAA). Because nursing students and faculty have been thrust into this environment, faculty must understand the Privacy Rule to ensure that both they and their students are in compliance. In this article, we provide a framework for faculty as they take a proactive leadership stance in relation to HIPAA. A multifaceted approach is necessary as faculty begin to navigate the challenges imposed by HIPAA. Strategies discussed include informing faculty and educating students, while developing a value system related to confidentiality issues and creating a safe environment for sharing information. PMID- 16342630 TI - An innovative model for nursing scholarship. AB - Ernest Boyer made a significant contribution to the manner in which scholarship is envisioned by faculty with his 1990 publication, Scholarship Reconsidered: Priorities of the Professoriate. The National League for Nursing Accreditation Commission (NLNAC) incorporated Boyer's scholarship principles into its 2002 accreditation criteria for nursing education programs. This article describes a model of scholarship developed by nursing faculty practicing in an undergraduate setting in a non-research-intensive institution. The model stems from their collegial work on defining scholarship in a way that is both professionally and personally satisfying. In addition, the model is congruent with the NLNAC's vision regarding nursing faculty scholarship. The model, called "Pillars of Scholarship," incorporates Boyer's four scholarship entities of discovery, application, integration, and teaching. PMID- 16342631 TI - Vulnerable bodies: competing discourses of intimate bodily care. AB - Nursing practices are subject to intersecting and competing discourses, which give them different, sometimes contradictory, meanings. In this article, using a case of intimate bodily care, we explore the workings of these discourses in a nursing student's account of her first experience of bathing an elderly woman. In particular we were interested in how different meanings of nursing "care" figure into the student's story. Through our analysis, we realized that a range of contradictions and tensions structured the student's experience. These contradictions were primarily produced by a collision between two discourses: the student's commonsense way of understanding the intimacy of such an act (liberal humanist discourse) and the impersonal approach characteristic of institutionalized care (scientific medical discourse). This collision caused distress and confusion, such that the student became almost as vulnerable as her patient. We argue that this type of detailed analysis of a nursing student's experience may increase our understanding of the discursive struggles that occur for students during their shifting between classrooms and clinical sites, between theory and practice, in the process of becoming professionals. Understanding these struggles and the relationships of power that produce both nursing student and patient as "vulnerable bodies" is important for nurse educators because they offer opportunities for intervention in the shaping of nursing professionals. PMID- 16342632 TI - Information literacy: developing lifelong skills through nursing education. AB - The amount and complexity of information nurses are expected to manage continues to increase exponentially. Support has grown for integrated curriculum approaches that include appropriate content on the use of a variety of information formats and instruction using resource-based and process methods. Such teaching-learning approaches demand a major shift in educational paradigms and encompass resource based learning, undergraduate research, service learning, inquiry learning, and problem-based learning. The implementation of an integrated curriculum promises advanced information skills, access, and use of available evidence to support clinical decision making and a foundation for lifetime learning. In this article, we argue that for information literacy to be enhanced, collaboration between teaching faculty and librarians must be fostered in meaningful ways. We report on the rationale of an integrated curriculum, changes to nursing education, and obstacles to the development and application of advanced information skills that exist within higher education and clinical settings. PMID- 16342633 TI - The effect of teaching method on long-term knowledge retention. AB - Choosing a teaching strategy that results in knowledge retention on the part of learners can be challenging for educators. Studies on problem-based learning (PBL) have supported its effectiveness, compared to other, more traditional strategies. The results of a previous study comparing the effect of lecture versus PBL on objective test scores indicated there was no significant difference in scores. To measure long-term knowledge retention, the same groups were evaluated 1 year after instruction. The posttest administered in the original study was repeated, and the scores from a comprehensive adult health examination and the endocrine subsection were analyzed. At an alpha level of 0.05, a statistically significant difference was found in the scores on two of the measures. The scores of the PBL group were significantly higher on the endocrine section of the examination and the repeat posttest. PMID- 16342634 TI - Incorporating research into associate degree nursing curricula. AB - All nurses are urged to be involved in research as participants and/or consumers. Such involvement may range from using research findings to participating in one or more of the research process steps to serving as principle investigator of a funded study. Clinicians at the bedside, including associate degree (AD) nurses, are well positioned to contribute to the body of nursing knowledge and enhance nursing care by identifying nursing care problems worthy of research, collecting data for nursing studies, and applying research findings in the care they provide. Minimal literature has addressed the preparation of AD students to assist in the research process. In this article, we describe how one AD program prepared students for their role as research participants. Content about the research process was presented in an easy-to-understand, clinically focused manner, and a brief research experience was provided to motivate and prepare these students for research involvement as nurses prepared at the AD level. The educational objectives were met, although time constraints limited students' opportunity to learn about research. PMID- 16342635 TI - Teaching undergraduate research from a process perspective. AB - Educators continue to struggle to find ways to teach nursing research to undergraduate students in a meaningful and engaging manner. The purpose of this article is to describe the development of an undergraduate nursing research course that incorporates a clinical practicum as a foundation for learning research concepts and the research process. By incorporating a clinical component into the research course, students more fully appreciate the connection between research and practice, more readily understand theoretical concepts discussed in class, and become more actively engaged in the learning process. The development of the collaborative clinical research project is described from a process perspective. PMID- 16342636 TI - Empowering nursing faculty and students for community service. AB - As the health care delivery system moves from acute care settings to community based services, nursing education must examine the experiences that increase students' abilities to function within a changing system. Currently, students receive community-based experiences that involve teaching health promotion concepts. However, students are not routinely prepared with specialized screening skills to use in the community. Using a train-the-trainer model, faculty and students were taught four specialized screening skills: Early and Periodic Screening, Diagnosis, and Treatment, Denver II Developmental Screening, vision screening, and hearing screening. Initially, faculty obtained certification as trainers for these specialized skills. Then, faculty educated students and provided enhanced, guided community experiences. Undergraduate, RN-to-BSN, and graduate family nurse practitioner students participated in this community-based experience. Project outcomes were evaluated based on the number of students and faculty who were certified and participated in these community activities. PMID- 16342637 TI - Interactive learning in an online nursing public policy course. PMID- 16342638 TI - Child development paper: theories and milestones in action. PMID- 16342639 TI - [Advantages and limits of nickel-titanium instruments for root canal preparation. A review of the current literature]. AB - In recent years, an important innovation in the field of dentistry has been the introduction of nickel-titanium alloy instruments. Superelasticity and shape memory are the main mechanical properties of this alloy which prompted its use in endodontics. Due to these characteristics, NiTi instruments have been demonstrated to preserve the original anatomy, the shape and position in space of the apical foramen. This literature review of mechanical properties of NiTi endodontic instruments aims to provide clinicians with a better understanding of their advantages and limits. PMID- 16342640 TI - [Atypical odontalgia]. AB - In spite of its first description by the English surgeon JOHN HUNTER more than 200 years ago, atypical odontalgia (AO), or phantom tooth pain, is not universally known among dentists. AO is a persistent neuropathic pain which may be initiated after deafferentiation of trigeminal nerve fibers following root canal treatment, apicectomy, or tooth extraction. In the absence of pathological clinical or radiological findings, the diagnosis is made by exclusion. After a thorough patient education about the condition, pharmacological and psychological pain management is required. Invasive and irreversible treatment attempts are contraindicated. PMID- 16342641 TI - [Therapeutic local anesthesia for the management of atypical odontalgia. A clinical case series]. AB - The management of patients diagnosed with atypical odontalgia (AO) is a challenging task. The aim of this longitudinal clinical case series was to document the effect of a 4% carticaine solution without epinephrine among ten patients with AO. Once or on two successive days, 1.7 ml of the local anesthetics was injected buccally in the area of the perceived pain. Pain intensity was recorded four times daily by each participant, beginning at least two weeks prior to the treatment and ending at least four weeks thereafter. The main outcome parameter was pain reduction (in percent) four weeks after treatment compared to the average pain intensity before the injection(s). In addition, study participants were asked to rate their satisfaction with the treatment. Four weeks after treatment, seven of the ten patients experienced significantly less pain than before. Among five individuals, there was a significant pain decrease (about 30% or more). Patient satisfaction was not significantly associated with the amount of pain reduction. Considering the limited treatment options and their side effects, injections with epinephrine-free carticaine seem to be a measure worth to be taken into consideration. Although results gained from uncontrolled case series have to be interpreted with caution, the findings of this observational study sound promising. PMID- 16342642 TI - [Milk fluoridation--where are we today?]. AB - This review deals with and discusses the history and objectives of milk fluoridation from its early beginnings in Winterthur/Switzerland up to current, WHO-sponsored controlled clinical trials with some 10000 children in several developing and emerging countries as well as in Great Britain. Today the fluoridation of milk in Switzerland does not represent an alternative to the fluoridation of salt and therefore has no practical meaning in this country. PMID- 16342643 TI - Dental fluorosis in salt fluoridation schemes. AB - The prevalences of fluoride-associated opacities (FOP) and nonfluoride-associated opacities (non-FOP) were assessed in two cantons of Switzerland with different salt fluoridation schemes. In the Canton of Zurich 305 schoolchildren from eight communities (age 9-10) were examined in 1995 and 900 children from one community (age 8-16) in 2002. In the Canton of Glarus 144 schoolchildren from 14 communities (age 9-10) were examined in 1996. In the Canton of Zurich 84% of the children used fluoridated domestic salt (250 ppm F) in the first five years of their life (questionnaire for parents 1995). In the Canton of Glarus, the corresponding value was 87%. In addition, about half of the bread samples in the Canton of Glarus showed an increased fluoride content (> 3 ppm F). Standardized photographs were taken of incisors dried with compressed air (15 seconds) and assessed using the Thylstrup Fejerskov index. The prevalence of FOP was 21% in eight communities of the Canton of Zurich (1995) and 22% in 14 communities of the Canton of Glarus (1996). No differences were found between these two locations, although fluoride intake was higher in the Canton of Glarus due to fluoridated salt used by bakeries. A recent examination in one community of the Canton of Zurich (2002) showed a lower prevalence of 11%. The prevalences of non-FOP in all locations were between 17% and 23%. The prevalences of FOP were at a low level in Switzerland compared to international studies using the same index. The most plausible explanation for this difference is the widespread use of low fluoride toothpastes (250 ppm F) by children under six. These toothpastes were introduced in 1986 and the consumption increased continuously until 1992. The low FOP prevalence cannot be considered a public health problem and is not perceived as such by the population. It is concluded that the Swiss fluoridation policy of the past two decades was approprate, with caries prevalence in permanent teeth of Swiss schoolchildren being among the lowest globally. PMID- 16342644 TI - [Structure and composition of the enamel in amelogenesis imperfecta]. AB - Amelogenesis imperfecta (AI) represents a broad spectrum of genetic diseases affecting the enamel formation. Suspected changes in the composition and structure of the enamel can hamper the establishment of an adequate adhesive bonding to the substrate, i.e. by influencing the conditioning. It was the purpose of this article to dwell on this problematic issue and to present a case with a hypomaturation type of AI. Using dual etch biopsies, the composition and structure of the affected enamel was studied and findings compared to healthy control teeth, which were extracted for orthodontic reasons. No differences in the calcium and phosphor content of the biopsies were noted. The Ca/P ratio was comparable as well. A sufficient etching pattern after phosphoric acid application was found. Clinical considerations and treatment concepts were discussed, which may also play a significant role when treating cases with other manifestations of enamel or dentin affections. PMID- 16342645 TI - [Injuries to permanent teeth. Part 2: Therapy of avulsion]. AB - The present paper describes the current treatment philosophy of dislocated or avulsed permanent teeth at our school. Splinting and medication (root canal treatment, systemic antibiotics, etc.) are targeted to minimize the initial inflammatory reaction and to reduce post-traumatic complications such as pulp necrosis and/ or external root resorption. PMID- 16342646 TI - Glycemic control in the intensive care unit: why we should wait for NICE-SUGAR. PMID- 16342647 TI - Avian H5N1 influenza--are we inching closer to a global pandemic? PMID- 16342648 TI - Influence of individual characteristics on outcome of glycemic control in intensive care unit patients with or without diabetes mellitus. AB - OBJECTIVE: To clarify the relationship of patient and critical illness characteristics (including any history of diabetes mellitus) to glycemic control with insulin and hospital mortality. PATIENTS AND METHODS: A case-control descriptive study was performed of patients admitted to a tertiary-care center multidisciplinary closed intensive care unit (ICU) at Mayo Clinic Hospital in Phoenix, Ariz, between January 1, 1999, and December 31, 2003, after implementation of a glycemic management protocol. Hospital mortality, the primary outcome, was examined in nondiabetic and diabetic ICU patients receiving insulin and in patients not requiring insulin (control group). RESULTS: Of 7285 patients, 2826 (39%) required insulin, 1083 of whom (15% of total) had a history of diabetes mellitus. The control group had a median (10th-90th percentile) glucose level of 118 mg/dL (range, 97-153 mg/dL) and a 5% mortality rate. The median glucose level was 134 mg/dL (range, 110-181 mg/dL) in nondiabetic patients and 170 mg/dL (121-238 mg/dL) in diabetic patients (P<.001), whereas mortality rates were 10% and 6%, respectively (P<.001). Compared with nondiabetic survivors, nondiabetic nonsurvivors had longer periods with glucose levels greater than 144 mg/dL. Diabetic nonsurvivors vs diabetic survivors had longer periods with glucose levels greater than 200 mg/dL. Poor glycemic control in nondiabetic patients was associated with increased insulin requirement and increased mortality. Critical illness characteristics that predicted poor glycemic control were advanced age, history of diabetes, cardiac surgery, postoperative complications, severity of illness, nosocomial infections, prolonged mechanical ventilation, or concurrent medications. CONCLUSIONS: Critical illness characteristics determined glycemic control and clinical outcome in ICU patients. Acute insulin resistance was associated with worse outcomes in nondiabetic patients. Although critical illness characteristics influenced glycemic control, future evaluation of the effect of insulin administration and optimal glycemic control in ICU patients is necessary. PMID- 16342650 TI - Weekly cyclophosphamide and alternate-day prednisone: an effective, convenient, and well-tolerated oral treatment for relapsed multiple myeloma after autologous stem cell transplantation. AB - OBJECTIVE: To assess the efficacy and tolerability of weekly oral cyclophosphamide in combination with alternate-day prednisone (CP) as salvage therapy for patients with relapsed multiple myeloma (MM) after autologous stem cell transplantation (ASCT). PATIENTS AND METHODS: We retrospectively reviewed the medical records of all patients identified in our clinical database as having received CP as treatment for relapsed MM after ASCT at Princess Margaret Hospital between July 1998 and May 2004. The CP regimen consisted of oral cyclophosphamide at 500 mg once weekly and oral prednisone at 100 mg on alternate days. RESULTS: A total of 66 patients received the CP regimen, with a median of 2.0 prior therapies (range, 1.0-5.0) from time of diagnosis to initiation of CP. The median time from relapse after ASCT to start of CP therapy was 1.5 months (range, 0.0 23.5 months). Because of nonsecretory disease in 7 patients, only 59 patients were evaluable for response. The median duration of CP treatment was estimated at 5.8 months (95% confidence interval [CI], 4.6-7.8 months). With a median follow up of 15.9 months (range, 1.4-67.2 months), 36 patients (61%) responded to treatment, 24 (41%) of whom had a partial response. The 1-year progression-free survival of all evaluable patients was estimated at 66% (95% CI, 54%-80%), with a median progression-free survival of 18.6 months (95% CI, 13.9-29.9 months). The median overall survival from time of initiation of CP was estimated at 28.6 months (95% CI, 22.1-not available months). CONCLUSION: Our data show that CP is an effective, well-tolerated, and convenient regimen as salvage therapy for MM after ASCT. PMID- 16342649 TI - The role of aspirin in the prevention of thrombotic complications of thalidomide and anthracycline-based chemotherapy for multiple myeloma. AB - OBJECTIVE: To study the efficacy of daily low-dose aspirin (81 mg orally) in decreasing the incidence of venous thromboembolic events (VTEs) in patients with multiple myeloma receiving pegylated doxorubicin, vincristine, and decreased frequency dexamethasone, plus thalidomide (DVd-T). PATIENTS AND METHODS: In this phase 2 clinical trial of DVd-T, conducted by the Cleveland Clinic Foundation from August 2001 to October 2003, 105 patients were enrolled. The first 35 patients experienced increased numbers of VTEs. von Willebrand levels and platelet aggregation to ristocetin before and after treatment with DVd-T increased significantly, suggesting a pathophysiology involving platelet endothelial interaction. Aspirin was added to the regimen, thus generating 3 patient groups: group 1 received aspirin from the start of DVd-T treatment before the study began (58 patients), group 2 received aspirin after the start of DVd-T treatment and after the study began (26 patients), and group 3 did not receive daily low-dose aspirin during the study (19 patients). Two patients being treated with warfarin for other indications were excluded from the study. The primary end point for this study was the incidence of VTE in the form of either deep venous thrombosis or pulmonary embolism. Secondary end points were the time to the first VTE, time to the composite end point of death or first VTE, and incidence of bleeding complications. RESULTS: After a median follow-up of 24 months, on an intent-to-treat basis, 26 posttreatment VTEs occurred after a median of 90 days, with 19% occurring in group 1, 15% in group 2, and 58% in group 3. Following multivariate time-to-event analysis, aspirin use continued to be associated with lower relative risk of VTE (hazard ratio, 0.22; confidence interval, 0.10-0.47; P<.001) and of the composite end point (hazard ratio, 0.28; confidence interval, 0.15-0.51; P<.001). CONCLUSION: Daily low-dose aspirin (81 mg orally) given to patients with newly diagnosed and relapsed/refractory multiple myeloma who were receiving DVd-T reduced the incidence of VTEs without an increase in bleeding complications. PMID- 16342651 TI - Risk factor implications of incidentally discovered uncomplicated bundle branch block. AB - OBJECTIVE: To evaluate the long-term outcome of a community-based patient population with incidentally discovered asymptomatic and uncomplicated bundle branch block (BBB). PATIENTS AND METHODS: A retrospective observational cohort study was undertaken of patients in Olmsted County, Minnesota, who were evaluated between 1975 and 1999 and were incidentally diagnosed as having BBB. We performed Kaplan-Meier analyses of all-cause mortality and development of first cardiac morbidity after the diagnosis of BBB, along with matched control group comparisons. RESULTS: A total of 723 patients with left BBB (LBBB) (58.1%) and right BBB (41.9%) met criteria. Mortality was higher in patients with BBB compared with controls (absolute difference of approximately 10% over 20 years; hazard ratio = 1.27; confidence interval, 1.02-1.58; P=.03) as was the development of first cardiac-related morbidity (hazard ratio = 1.32; confidence interval, 1.14-1.54; P<.001). Patients with BBB and without the risk factors of diabetes, hypertension, and/or hypercholesterolemia showed increased long-term mortality compared with matched controls (no BBB) also without risk factors (P=.02). However, comparable mortality was shown between patients with BBB who did not have these risk factors and matched control patients who had these risk factors. The risk of developing cardiac-related morbidity also was increased in the presence of BBB, particularly LBBB. CONCLUSIONS: Uncomplicated asymptomatic BBB (notably LBBB) with normal left ventricular ejection fraction is not benign. Our findings indicate that the presence of isolated BBB denotes a high-risk patient subgroup that has a compromised long-term outcome comparable to patients with conventional cardiovascular risk factors. PMID- 16342652 TI - Coronary heart disease in ethnically diverse women: risk perception and communication. AB - OBJECTIVES: To assess perceived vs calculated risk of coronary heart disease (CHD), preferred methods of communicating risk, and the effect of brief educational intervention to improve accurate perceptions of personal risk. SUBJECTS AND METHODS: Of 1858 women who underwent screening mammography between April and September 2003 at the Columbia University Medical Center in New York, NY, we assessed 125 women with no history of cardiovascular disease who participated in a risk factor screening and education program. Demographic variables were evaluated by interviewer-assisted standardized questionnaires. Absolute 10-year CHD risk was calculated using the Framingham global risk assessment. Perceived 10-year risk and preferred method of communicating risk were evaluated systematically. RESULTS: Among 110 research participants who were eligible for risk estimation, 59% had a 10-year risk of less than 10%. However, only half of those women accurately perceived their risk as low. After a brief educational intervention, the women's ability to correctly categorize their personal CHD risk improved significantly. Preferred methods to communicate risk varied by level of education and age. Older women (> or = 65 years) and those with a high school education or less were more likely to prefer simple methods of having CHD risk communicated compared with their counterparts. CONCLUSIONS: These data underscore the need to determine preferences for providing risk information and to test various formats for communicating CHD risk to improve awareness and management of CHD risk factors, especially among women of different age groups and education levels. PMID- 16342653 TI - Five-year prospective study of the incidence and prognosis of peripartum cardiomyopathy at a single institution. AB - OBJECTIVE: To determine the incidence and prognosis of peripartum cardiomyopathy (PPCM) in rural Haiti. PATIENTS AND METHODS: Prospectively identified patients with PPCM treated at the Hospital Albert Schweitzer (HAS), Deschapelles, Haiti, were included in this study. Patients who presented to HAS from February 1, 2000, to January 31, 2005, were identified through a search of the HAS PPCM Registry. Clinical and serial echocardiographic data were collected on these patients. RESULTS: The 5-year experience confirms the high incidence of PPCM in this area, approximately 1 case per 300 live births, which is severalfold the estimated incidence in the United States (estimated 1 case per 3000 to 4000 live births). In this population, the ratio of PPCM deaths for the 5-year period was 47.1 per 100,000 births compared with the US ratio of 0.62 per 100,000 births. The mortality rate was 15.3% (15 deaths of 98 patients), and the mean follow-up was 2.2 years (range, 1 month to 5 years). Five years after the initiation of the HAS PPCM Registry search, 26 (28%) of 92 patients with PPCM observed for at least 6 months had regained normal left ventricular function. The difference in left ventricular echocardiographic features at diagnosis between deceased patients and survivors was not statistically significant: mean end-diastolic dimension (6.2 vs 5.8 cm; P=.08), ejection fraction (22% vs 25%; P=.12), and fractional shortening (16% vs 15%; P=.46). Left ventricular echocardiographic features at diagnosis were unable to predict individually who would eventually recover, although a statistically significant difference occurred at diagnosis between the recovered group and nonrecovered group for mean ejection fraction (28% vs 23%; P<.001) and fractional shortening (17% vs 14%; P=.004). CONCLUSION: Peripartum cardiomyopathy occurs significantly more commonly in rural Haiti on a per capita basis than in the United States. Patients with PPCM have a higher mortality rate and a poorer return of normal ventricular function. PMID- 16342654 TI - Oxidative stress and hematologic and biochemical parameters in individuals with Down syndrome. AB - OBJECTIVE: To evaluate the levels of thiobarbituric acid reactive substances (TBARS), uric acid, and seric superoxide dismutase and catalase activities, as well as total serum iron, total iron-binding capacity (TIBC), erythrocyte osmotic fragility, and hemograms in people with Down syndrome. PARTICIPANTS AND METHODS: The study sampled (July to December 2003) 50 people with Down syndrome and 50 individuals without Down syndrome (control group) matched by age and sex. The levels of TBARS were measured by the TBARS method. Serum superoxide dismutase and catalase activities and uric acid levels were determined spectrophotometrically. Erythrocyte osmotic fragility was assessed by the percentage of hemolysis. Hemograms, total serum iron level, and TIBC were determined with automated systems. RESULTS: The results showed that levels of TBARS, uric acid, and seric superoxide dismutase and catalase activities were higher in the Down syndrome group compared with the control group. We also observed a slight increase in erythrocyte osmotic fragility in the Down syndrome group, but the total serum iron levels, TIBC, and hemograms for both groups were within the age-related reference values. CONCLUSION: This was the first time, to our knowledge, that increases in seric superoxide dismutase and catalase activities were observed in people with Down syndrome. Although other studies are necessary, our results add to the understanding of the mechanisms responsible for the increased oxidative stress observed in individuals with Down syndrome and may be useful in supporting future antioxidant therapies that will improve the lives of people with Down syndrome. PMID- 16342655 TI - Medical student distress: causes, consequences, and proposed solutions. AB - The goal of medical education is to graduate knowledgeable, skillful, and professional physicians. The medical school curriculum has been developed to accomplish these ambitions; however, some aspects of training may have unintended negative effects on medical students' mental and emotional health that can undermine these values. Studies suggest that mental health worsens after students begin medical school and remains poor throughout training. On a personal level, this distress can contribute to substance abuse, broken relationships, suicide, and attrition from the profession. On a professional level, studies suggest that student distress contributes to cynicism and subsequently may affect students' care of patients, relationship with faculty, and ultimately the culture of the medical profession. In this article, we review the manifestations and causes of student distress, its potential adverse personal and professional consequences, and proposed institutional approaches to decrease student distress. PMID- 16342656 TI - Aldosterone receptor antagonists in the medical management of chronic heart failure. AB - The benefits of aldosterone receptor antagonists (spironolactone and eplerenone) for patients with heart failure were shown in 2 recent randomized controlled trials. Some of the proposed mechanisms of action of aldosterone antagonists are (1) inhibition of myocardial and vascular remodeling, (2) blood pressure reduction, (3) decreased collagen deposition, (4) decreased myocardial stiffness, (5) prevention of hypokalemia and arrhythmia, (6) modulation of nitric oxide synthesis, and (7) immunomodulation. Like many hormone receptors, the aldosterone receptor can be either nuclear or membrane bound. Most of the activities of the aldosterone receptor are subserved by the nuclear receptors and often lead to alterations in gene transcription. Although these agents are well tolerated in carefully selected patient populations that meet the inclusion criteria of large clinical trials, their use in unselected elderly patients with heart failure and multiple comorbidities has been associated with a significant risk of hyperkalemia and renal failure. Although no convincing data exist to predict which individual patients will respond to aldosterone inhibition, patients with more severe heart failure and those with acute myocardial infarction with concomitant heart failure or left ventricular dysfunction are most likely to respond. Theoretically, aldosterone receptor antagonists may also be beneficial in patients with more mild to moderate systolic heart failure or even in those with diastolic heart failure, although direct evidence is still lacking. PMID- 16342657 TI - Tracheostomy in critically ill patients. AB - Tracheostomy is a common critical care procedure in patients with acute respiratory failure who require prolonged mechanical ventilatory support. Tracheostomy usually is considered if weaning from mechanical ventilation has been unsuccessful for 14 to 21 days. A recent clinical trial suggested that early tracheostomy may benefit patients who are not improving and who are expected to require prolonged respiratory support. In this study, early tracheostomy improved survival and shortened duration of mechanical ventilation. Minimally invasive bedside percutaneous tracheostomy was introduced recently as an alternative to the traditional surgical technique. In expert hands, the 2 techniques are equivalent in complications and safety; however, the bedside percutaneous approach may be more cost-effective. Tracheostomy should be considered early (within the first week of mechanical ventilation) in patients with a high likelihood of prolonged mechanical ventilation. Depending on local medical expertise and costs, either the percutaneous or the surgical technique may be used. PMID- 16342658 TI - Health care maintenance in female adolescents. AB - The assessment and care of female adolescents by primary care physicians can be facilitated with increased knowledge about this stage of development, the health care risks faced by these patients, and the resources available to aid in their care. With a focus on preventive health maintenance, this concise review addresses these areas as well as how to build relationships with female adolescent patients, conduct age-appropriate interviews and tests, and maintain patient confidentiality. PMID- 16342659 TI - 75-year-old man with progressive shortness of breath on exertion. PMID- 16342661 TI - Chronic lymphoid leukemias other than chronic lymphocytic leukemia: diagnosis and treatment. AB - The World Health Organization classification divides lymphoid malignancies into precursor B-cell and T-cell neoplasms as well as mature B-cell and T-cell neoplasms. Mature B-cell neoplasms comprise more than 85% of non-Hodgkin lymphomas worldwide and can be further subclassified according to the postulated cell of origin by using specific morphologic, immunophenotypic, and molecular characteristics. Similarly, the more uncommon mature T-cell neoplasms have been better characterized to include numerous distinct entities with widely varying natural histories. The distinction between lymphoma and leukemia is somewhat arbitrary and is based on variable involvement of the bone marrow, peripheral blood, and lymphatic system. In this article, we review the diagnostic and clinical features of mature B-cell and T-cell lymphoproliferative disorders that commonly have a leukemic presentation. PMID- 16342660 TI - Spurious hypocalcemia after gadodiamide administration. AB - Severe hypocalcemia may require prompt intervention to avoid life-threatening consequences. We report a case in which a 78-year-old man had a critically low serum calcium level measured with use of standard colorimetric assay after gadodiamide administration during magnetic resonance angiography. Reanalysis of the same serum specimen using absorption spectroscopy revealed normal calcium values, confirming the diagnosis of spurious hypocalcemia. The increasing use of gadolinium chelates during magnetic resonance imaging and anglography will lead to a marked increase in reports of critically low serum calcium values. Increasing physicians' awareness of gadodiamide-induced spurious hypocalcemia may prevent unnecessary and potentially inappropriate therapeutic interventions. PMID- 16342662 TI - [Gene and cell therapy approaches to cardiovascular and systemic diseases in modelic experiments on animals]. AB - The therapeutic potential of gene and cell therapy in cardiovascular and systemic disease such as myocardial ishemia, postangioplasty restenosis, hypertensia, arteriosclerosis, trombogenicity, posttransplantation complications is considered. PMID- 16342663 TI - [Role of membrane deformations in regulation of functions of cells]. AB - There is evidence that cells respond not only to biologically active substances, but also to external mechanical effects. This is especially important for cellular elements of vascular walls and blood. Mechanical treatment cause so called stretch-deformation of plasma membranes and forces generated by shift of plasma layers induce shear stress. Good experimental evidence exists that mechanical effects influence functional activity of vascular wall and blood cells. PMID- 16342664 TI - [Biological role of carnosine and its use in ophthalmology (mini-review)]. AB - Carnosine (beta-alanyl-L-histidine) exhibits various biological effects underlying its potential therapeutic use. In ophthalmology carnosine was effective in treatment of corneal metabolic diseases, senile cataract. It accelerated cornea reepithelization after photorephraction ceratoectomia. PMID- 16342665 TI - [Lysosomal cysteine proteinases at neoplastic transformation]. AB - This review summarizes the data on classification, properties, structure and biological functions of mammalian papain-like lysosomal cysteine proteinases (cathepsins). The most studied are cathepsins B, H, L, S and K. The peculiarities of lysosomal cysteine proteinase regulation, particularly regulation by specific endogenous inhibitors are discussed. Involvement of these enzymes in development of physiological and pathological processes are considered. Special attention is paid to the role of cysteine proteinases and their inhibitors in neoplastic transformation. The possibility of use cathepsin endogenous inhibitors as basis for design antitumorogenic therapeutic substances are discussed. PMID- 16342666 TI - [Optical biosensor study and molecular modeling of interactions between cytochrome P450cam and cytochrome b5]. AB - The formation of complexes of cytochrome P450cam (P450cam) with full-length cytochrome b5 (d-b5) and its tryptic water-soluble fragment (t-b5) was analyzed using a two-channel IAsys+ optical biosensor. It was found that t-b5 can form complexes with P450cam, while d-b5 does not interact with P450cam. The involvement of amine groups of P450cam in the complex formation was demonstrated. The temperature dependence of t-b5(im)/P450cam complex formation was measured. The association rate constant (k(on)) increased with temperature, while the dissociation rate constant (k(off)) practically remained unchanged. It was concluded that hydrophobic interactions play a key role in the complex formation, while electrostatic interactions are significant for complex stabilization. Based on temperature dependence the activation energy, enthalpy and entropy of complex formation were calculated. It was shown that the entropy component plays a key role in t-b5(im)/P450cam interaction. Computer modeling of P450cam/t-b5 and P450cam/d-b5 interactions was carried out. Using the method of molecular docking some hypotheses of protein-protein complexes were advanced and the best ones were selected based on geometric complementarity, calculated binding energy and probability of electron tunneling between proteins. The computer modeling has shown that only P450cam and t-b5 can form the stable complex. These results are in good agreement with the experimental data obtained with the optical biosensor. PMID- 16342667 TI - [The effect of Asian cobra venom on the content of rat heart and liver mitochondrial phospholipids and the protective effect of benzonal]. AB - Administration of a cobra venom to rats resulted in decrease of heart and liver mitochondrial cardiolipins (1.88 and 1.82 times), phosphatidylethanolamines (1.56 and 1.42 times) and phosphatidic acid (1.14 and 1.29 times), whereas the content of phosphadtidylcholins increased by 1.42 and 1.51-fold, respectively. The content of phosphadtidylinositol increased by 3.09 times in liver mitochondria and decreased by 1.28 times in heart mitochondria. Administration of benzonal not only normalized liver mitochondrial phosphadtidylserine content impaired after the venom administration but even exceeded it. Benzonal also minimized changes in mitochondrial phospholipid induced by the cobra venom, and also reduced liver and heart phosphatidic acid content by 1.36 and 1.7 times. PMID- 16342668 TI - [The influence of sodium nitroprusside, the possible donor of nitric oxide on activity ATPases and lipid peroxidation of erythrocytes of sick children]. AB - The effect of sodium nitroprusside, possible nitric oxide donor, on the activities of Ca2+, Mg(2+)- and Na+, K(+)-ATPases and lipid peroxidation was investigated in erythrocytes in vitro. Sodium nitroprusside caused 3-fold increase of mechanical and peroxidative erythrocyte hemolysis and also increase of bound malonic dialdehyde. Sodium nitroprusside-treated erythrocytes were also characterized by increased Na+, K(+)-ATPase activity. PMID- 16342669 TI - [Adenine nucleotide system in adult and old rat liver during immobilization stress]. AB - Adenine nucleotide and inorganic phosphate concentrations were studied in livers of adult (10-12 month) and old (22-25 month) Wistar male rats during immobilization stress. Study revealed that level of liver energy provision supply of old rats was lower than in adults. Stress caused reduction of adenine nucleotide concentration in the liver of both adult and old animals. PMID- 16342670 TI - [Noninvasive prenatal gender determination by means of PCR]. AB - Prenatal diagnostics of genetic diseases becomes more and more popular. Classic obstetric approach for diagnostics of numerous genetic diseases employs biopsy or amniotic liquid analyses. Good evidence now exists that polymerase chain reaction (PCR) is one of the most powerful tools of prenatal diagnostics. In contrast to ultrasound investigation PCR is absolutely safe for an embryo and is much more sensitive at early stage of gestation. PCR analysis can recognize male fetal DNA in mother blood and detect some gender-related genetic diseases. Using detection of Y-chromosome in peripheral blood we have analyzed a diagnostic value of some markers sites of Y-chromosome during gestation, type of blood sample (whole blood, plasma or serum) and varioations of the PCR-method (single-step PCR or nested PCR). Comparative analysis of DNA sequences using NCBI Blast we have found Y-chromosome sites (loci DYS14 and ZFY) suitable for PCR identification of male DNA. Blood plasma is the most optimal blood sample for PCR prenatal gender determination. Prenatal gender determination by PCR can be diagnosed at 4-6 weeks gestation. PMID- 16342671 TI - [Peculiarities of free radical and antioxidant statute in patients with systemic lupus erythematosus and rheumatoid arthritis]. AB - The state of lipid peroxidation and antioxidant defence was investigated in sera of 66 patients with rheumatoid arthritis (RA), 50 patients with systemic lupus erythematosus (SLE) and in 235 healthy donors using the method of chemiluminescence and EPR-spectroscopy. The results indicate involvement of free radical oxidation in pathogenesis of RA and SLE. Peroxide and antioxidant imbalance was more pronounced in RA than in SLE. PMID- 16342672 TI - [Total antioxidant capacity of biological fluids for chronic kidney insufficiency using coulometric titration data]. AB - Express coulometric method for estimation of blood and plasma antioxidant capacity based on the use of electrogenerated compounds of bromine is proposed. Using this method we found changes in total antioxidant capacity in patient with chronic kidney insufficiency during hemodialysis session and after it. PMID- 16342673 TI - [Elevated plasma homocysteine and glutathione level in patients with renal failure]. AB - The levels of homocysteine (Hcy) and glutatione (Glt) were determined in plasma of 219 patients with different stages of chronic kidney disease (CKD) (94 patients received chronic bicarbonate hemodialysis and 125 patients were at predialysis stage). The elevated Hcy level was detected in an early stage of CKD and reached maximal values and spreading in the hemodialysis patients. The levels of total and reduced Hcy (tHcy and rHcy, respectively) were also higher in CKD patients: the proportion of rHcy in total pool of plasma Hcy was significally higher than in healthy subjects. In the group of non-dialysed patients the reverse dependance between glomerular filtration rate (GFR) value and Hcy plasma concentration was found. We did not find any correlation between Glt and GFR in predialysis patients, but there was direct dependance between Glt and tHcy plasma levels in this group. Glt plasma levels in dialysis patients were reliably lower than in predialysis ones. There was a lack of some correlation between Glt and tHcy in the haemodialysis group. PMID- 16342674 TI - [Current status and prospect of metabonomics]. PMID- 16342675 TI - [N-myristoyltransferase: a novel antifungal target]. PMID- 16342676 TI - [Percutaneous penetration of ketoprofen and ketoprofen isopropyl ester through a tissue engineering skin reconstructed with HaCaT cells]. AB - AIM: To reconstruct of a tissue engineering skin in vitro for the study of the use of drug percutaneous penetration and metabolism. METHODS: Dermal fibroblasts were embedded in collagen type I. HaCaT cells were seeded on the top of the gel. The skin was generated through air-liquid interface culture. Effects of various culture media on tissues morphology were investigated. Sections of the cultured skin were stained with hematoxylin and eosin and examined under microscope. Permeation and metabolism of ketoprofen and its isopropyl ester through the cultured skin were investigated. RESULTS: HaCaT cells initially developed a multilayer epithelium at the air-liquid interface, but it showed a parakeratotic stratum corneum. Vitamin C enhanced cell proliferation obviously. Vitamin D3 promoted cell differentiation. And estradiol showed little effect on the tissue engineering skin. Ketoprofen isopropyl ester was hydrolyzed into ketoprofen when penetrated through the cultured skin, which resembled in the skin cell homogenates metabolism. CONCLUSION: Cultured at the air-liquid interface, HaCaT cells developed a parakeratotic mutilayer epithelium. Enzyme activity was reserved. This cultured skin could serve as an appropriate model for drug percutaneous metabolism and skin irritation. PMID- 16342677 TI - [In vitro and in vivo pharmaceutical behaviors of lycopene microcapsules]. AB - AIM: To evaluate in vitro release of lycopene microcapsules. Pharmacokinetic parameters of lycopene microcapsule and lycopene powder as reference were estimated after a single dose of oral administration to dogs. The relationship between in vitro dissolution and in vivo absorption was investigated. METHODS: The content of lycopene in the release medium was determined by UV spectroscopy method. Health hybrid male dogs were used as experiment subjects and lycopene powder used as standard to estimate the pharmacokinetics of lycopene microcapsules. HPLC method was used to assay the concentration of lycopene in dog plasma. Pharmacokinetics parameters were estimated by 3P87 program. The drug release percentage in stimulated intestinal fluid was compared with the absorption at a given time point. RESULTS: The release profiles of lycopene from microcapsule showed that the lycopene gelatin microcapsule exhibited enteric property. The pharmacokinetics parameters estimated after oral administration of lycopene powder and lycopene microcapsule in a single dose of 2.5 mg x kg(-1) body weight to dogs were 7.30 h, 15.06 h for T1/2alpha; 28.10 h, 46.76 h for T1/2beta; 22.32 h, 41.03 h for T(max); 1.67 microg x h x L(-1), 2.08 microg x h x L(-1) for AUC(0-infinity), respectively. The concentration-time curves could be fitted to a two-compartment model for both the lycopene powder and the lycopene microcapsule analyzed by 3P87 program. The relationship between in vitro dissolution and in vivo absorption was found to have good correlation (r = 0. 981 9) was found. CONCLUSION: It could be concluded that lycopene microcapsule was a sustained release dosage form. The result of release in vitro could be used to predict the absorption in vivo. PMID- 16342678 TI - [Identification of ligands for human LOX-1 through fluorescence polarization based high throughput screening]. AB - AIM: To develop a fluorescence polarization-based high throughput screening and identify ligands for human Lectin-like oxidized low-density lipoprotein receptor 1 (hLOX-1). METHODS: Sequential ultracentrifugation at 4 degrees C from normolipidemic fasting volunteers to obtain low density lipoprotein (LDL), which was modified by CuSO4 (5 micromol x L(-1)) at 37 degrees C for 24 h. The assay was based on the interaction between receptor and ligand, and hLOX-1 was labeled by FITC and bound to its specific ligand, oxLDL. Different reaction time and DMSO concentration were optimized to determine the stability and tolerance of fluorescence polarization (FP) assay. 3 200 compounds were screened in black 384 well microplate by FP-based competitive displacement assay, at excitation filter of 485 nm and emission filter of 530 nm. Z' was used to assess the assay quality. RESULTS: The FP-based HTS was formatted in a 384-well microplate with a Z' factor of 0. 75, and three active compounds for hLOX-1 were identified with IC50 below 40 micromol x L(-1) from total 3 200 compounds. CONCLUSION: The results indicated that the fluorescence polarization assay is stable, sensitive, reproducible and well suited for high throughput screening efforts. PMID- 16342679 TI - [Impact of cytochrome P450 CYP2C9 variant allele CYP2C9 * 3 on the pharmacokinetics of glibenclamide and lornoxicam in Chinese subjects]. AB - AIM: To investigate the impact of CYP2C9 * 3 on the pharmacokinetics of glibenclamide and lornoxicam. METHODS: CYP2C9 * 3 was measured in 83 non-related Chinese subjects by PCR-RFLP. The pharmacokinetics of lornoxicam and glibenclamide were investigated in 18 subjects (7 with CYP2C9 * 1/* 3 genotype and 11 with * 1/* 1 genotype). Glibenclamide and lornoxicam in plasma were determined by the sensitive liquid chromatography-tandem mass spectrometry, separately. RESULTS: After a single oral dose of 2.5 mg glibenclamide, C(max) was (70.0 +/- 11.5) microg x L(-1) in CYP2C9 * 1/ * 3 subjects and (51.9 +/- 12.3) microg x L(-1) in * 1/ *1 subjects. AUC(0-infinity) were (435 +/- 47) vs (287 +/- 95) microg x h x L(-1) (in * 1/ * 3 vs * 1/ *1 subjects), and CL/F were (96 +/- 9.3) vs (160 +/- 51) mL x min(-1), respectively. Statistic analysis results indicated that glibenclamide AUC(0-infinity) was significantly higher (1.5-fold) and subsequently CL/F was significantly lower (40%) in CYP2C9 * 1/ * 3 subjects than those in * 1/ * 1 subjects (P < 0.01). After a single oral dose of 8 mg lornoxicam, C(max) was (1.54 +/- 0.24) mg x L(-1) in CYP2C9 * 1/ * 3 subjects and (1.19 +/- 0.37) mg x L(-1) in * 1/ * 1 subjects. AUC(o-infinity were (14.9 +/- 2.2) vs (6.92 +/- 1.48) mg x h x L(-1) (in * 1/ *3 vs * 1/ * 1 subjects), and CL/F were (9.1 +/- 1.2) vs (20.1 +/- 4.6) mL x min(-1), respectively. Statistic analysis results indicated that lornoxicam AUC(0-infinity) was significantly higher (2. 2-fold) and subsequently CL/F was significantly lower (55% ) in CYP2C9 * 1/ * 3 subjects than those in * 1/ * 1 subjects (P < 0.001). CONCLUSION: CYP2C9 * 3 greatly affects both the pharmacokinetic profiles of glibenclamide and lornoxicam. The elimination of these drugs significantly decreased in subjects with CYP2C9 * 1/ * 3 genotype, especially lornoxicam. PMID- 16342680 TI - [Inhibition of the expression of prostate specific antigen by curcumin]. AB - AIM: To study the effect of curcumin on the expression of prostate specific antigen (PSA). METHODS: AXSYM system-chemical luciferase method was used to examine the content of PSA in prostate cancer cell lines, LNCap after treated with different doses of curcumin. pGL3-PSA luciferase expression vector, containing 640 bp DNA of PSA gene 5' promoter region was constructed and transfected into LNCap cell with lipofectin. Through detecting the activity of luciferase, the effect of curcumin on the promoter of PSA was studied. Western blotting was used to detect expression of androgen receptor (AR) in LNCap cell with different concentrations of curcumin. RESULTS: The expression of PSA was inhibited and activity of luciferase was reduced by curcumin. There was also significant difference in AR expression as shown by Western blotting experiment after treatment of different doses of curcumin. CONCLUSION: Through inhibiting AR expression, curcumin reduced the function of PSA promoter and inhibited PSA protein expression. PMID- 16342681 TI - [Emodin blocks voltage dependent potassium channels in rat proximal colon smooth muscle cells]. AB - AIM: To investigate the effect of emodin on the voltage dependent potassium (K(V)) currents in rat proximal colon smooth muscle cells. METHODS: Whole cell patch clamp technique was used to record potassium currents including fast transient outward current (I(KA)) and delayed rectifier current (I(Kdr)). Contamination of calcium-dependent potassium currents was minimized with CdCl2 in external solution and EGTA in pipette solution. RESULTS: Emodin (1-30 micromol x L(-1)) reversibly and dose-dependently reduced the amplitude of I(Kdr) with an K(d) value of (1.9 +/- 0.1) micromol x L(-1). I(KA) was also inhibited with 30 micromol x L(-1) emodin to a lesser extent. Although acceleration of the decay rate of the K(V) currents was observed, the block by emodin was not through open block mechanism because a steady state level of inhibition of I(Kdr) was achieved during the first pulse from holding potential -70 mV to + 50 mV after the cells were holding at -70 mV for a three minutes interval in the presence of emodin. Emodin (5 micromol x L(-1)) had no effect on the steady-state activation and inactivation kinetics of K(V) currents, but 30 micromol x L(-1) of emodin produced a positive shift of the voltage dependence of activation, and an increase in the steepness of activation gating as well as shifted the voltage dependence of inactivation to positive direction. CONCLUSION: Emodin, not through open block mechanism, markedly reduced the amplitude of I(KA) and I(Kdr) and modulated the gating properties of K(V) channels in a reversible and dose dependent manner. PMID- 16342682 TI - [Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cell]. AB - AIM: To investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line. METHODS: The HPDE3A cDNA was recombined with baculovirus, and then the recombinant was transfected into Tn cell line. The expression of HPDE3A in Tn cell line was detected and identified by the RT-PCR, SDS-PAGE and Western blotting. RESULTS: The recombinant HPDE3A protein was stably expressed in Tn cell line and detected by the distinct morphological changes of Tn cell, RT-PCR, SDS-PAGE and Western blotting using polyclonal antibody. The M(w) of the recombinant protein was about 120 kD. CONCLUSION: Recombinant HPDE3A can be expressed in Tn cell line using the baculovirus expression system, and thus provided the basic material for studying its bioactivity and application in screening for HPDE3A inhibitor. PMID- 16342683 TI - [Design and synthesis of quinoxaline derivatives and their antitumor activities]. AB - AIM: To design and synthesize novel quinoxaline derivatives as antitumor agents. METHODS: Using 4-chloro-2-nitroaniline as a starting compound, followed by substitution, reductive cyclization, oxidation, and chlorination, to give the key intermediate 2,7-dichloroquinoxaline (7), which reacted with different phenolic compounds to afford quinoxaline derivatives. RESULTS: The structures of the target molecules were characterized by elemental analysis, 1H NMR, MS, and IR. CONCLUSION: At concentration of 1 x 10(-4) mol x L(-1), some of the derivatives showed equal antitumor activities to XK469. PMID- 16342684 TI - [Prodrug structural modifications of cyclovirobuxine D and their biological activity]. AB - AIM: To search for compounds for the treatment of cardiovascular diseases through prodrug structural modifications of cyclovirobuxine D, a single efficient composition distilled from Box plant in China, which was used to treat angina and myocardial infarction. METHODS: According to prodrug design principle, a series of cyclovirobuxine D analogues were prepared, suc as succinate, phosphate and amino acid ester, and their biological activities were tested. RESULTS: Seven new compounds were obtained and confirmed with 1H NMR, MS, and element analysis. CONCLUSION: In pharmacology experiment, for treating arrhythmia induced by aconitine, succinate and amino acid ester of cyclovirobuxine D (I and VII) showed better activities than that of cyclovirobuxine D. The normal rhythm of the heart duration of I and VII were ( 11.53 +/- 7.62) min and (12.68 +/- 9.25) min, compared with 0.9% NaCl solution and cyclovirobuxine D, (2.36 +/- 1.68) min and (10.25 +/- 6.59) min (P < 0.01), respectively. Another pharmacology experiment, for treating arrhythmia induced by chloroform, the negative ratio of I and VII were 80% and 82%, compared with 0.9% NaCl solution and cyclovirobuxine D, 43% and 52% (P < 0.05), respectively. The difference between new compounds and cyclovirobuxine D was distinct. PMID- 16342685 TI - [Synthesis of a novel L-nucleoside, beta-L-D4A and its inhibition on the replication of hepatitis B virus in vitro]. AB - AIM: Nucleoside analogues have become the most promising candidates of anti-HBV drugs. In this study, beta-L-D4A was synthesized and explored its inhibitiory action against hepatitis B virus (HBV) in 2. 2. 15 cells derived from HepG2 cells transfected with HBV genome. METHODS: beta-L-D4A was stereo-controlled synthesized from D-glutamic acid, and the structure was identified by IR, 1H NMR and MS. 2. 2. 15 Cells were placed at a density of 5 x 10(4) per well in 12-well tissue culture plates, and treated with various concentrations of beta-L-D4A for 6 days. At the end, medium was processed to obtain virions by a polyethlene glycol precipitation method. At the same time, intracellular DNA was also extracted and digested with Hind III. Both of the above DNA were subjected to Southern blot, hybridized with a 32P-labeled HBV probe and autoradiographed. The intensity of the autoradiographic bands was quantitated by densitometric scans of computer and EC50 was calculated. 2. 2. 15 cells were also seeded in 24-well tissue culture plates, and cytotoxicity with different concentrations was examined by MTT method. IC50 was calculated. RESULTS: The synthesized compound structure conformed with beta-L-D4A; Autoradiographic bands showed similar for supernatant and intracellular HBV DNA. Episomal HBV DNA was inhibited in a dose dependent manner. EC50 0.2 micromol x L(-1). The experiment of cytotoxicity gained IC50 200 micromol x L(-10. CONCLUSION: beta-L-D4A has been synthesized successfully. beta-L-D4A possessed potent inhibitory effect on replication of HBV in vitro with low cytotoxicity, TI value was 1 000. It is expected to be developed clinically into a new anti-HBV drug. PMID- 16342686 TI - [Isolation and identification of two new epimer from the mother liquid of megestrol acetate]. AB - AIM: To study the impurity in the drug megestrol acetate. METHODS: Chromatography methods were used to separate the chemical constituents. Their structures were determined by NMR and MS spectral analysis. RESULTS: Two new epimers were isolated from the mother liquid of the drug megestrol acetate. CONCLUSION: These new epimers were identified as 17alpha-acetoxy-2beta,6alpha-dimethylprega-4-ene 3,20-dione (1) and 17alpha-acetoxy-2alpha,6alpha-dimethylprega-4-ene-3,20-dione (2). PMID- 16342687 TI - Biotransformation of 6beta-santonin by Phytolacca acinosa cell suspension cultures. AB - AIM: To obtain more valuable derivatives for the further structural modification of 6beta-santonin (1) via biotransformation by using cell suspension cultures of Phytolacca acinosa. METHODS: The substrate 1 was incubated with cell suspension cultures of P. acinosa, the products were obtained by chromatography, and identified on the basis of their physical and spectral data (HRMS, 1D NMR, 2D NMR, NOE and IR). RESULTS: After incubation with cell suspension cultures of P. acinosa, 1 was converted into five products. Among them, 3 is a new compound. CONCLUSION: 6beta-santonin could be selectively reduced and hydroxylated by the cell suspension cultures of P. acinosa, which would provide valuable intermediates for its further structural modification. PMID- 16342688 TI - [Analysis of ephedrine and its metabolites in rat urine by HPLC-ESI-ITMSn]. AB - AIM: To estabilish a rapid and sensitive LC-ESI-ITMSn method for the identification of ephedrine and its main metabolites in rat urine. METHODS: After optimizing the detection condition of LC-ESI-ITMSn chromatography and mass spectrometry by using a standard ephedrine, the ionization and cleavage rules of ephedrine in ESI-MS and ESI-MSn modes were summarized, and then serving as the basis for the metabolite analysis of ephedrine in rat urine. Rat urine samples of 0-48 h were collected after ig 10 mg x kg(-1) ephedrine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC-ESI-ITMSn. RESULTS: The structures of ephedrine metabolites were elucidated according to the changes of the molecular weights of the metabolites (deltaM) and their cleavage pattern in ESI-ITMSn. As a result, three phase I metabolites and the parent drug ephedrine were identified existing in rat urine, but no phase II metabolites were found. CONCLUSION: The LC-ESI-ITMSn method is rapid and highly sensitive and sepecific, it is suitable for the identification of ephedrine and its metabolites in rat urine. PMID- 16342689 TI - [Application of HPLC/ELSD fingerprint and constellation graphical clustering method applied to evaluate the quality of Qingkailing injection]. AB - AIM: To evaluate the quality of Qingkailing injections effectively, and to develop a credible method for the quality control of traditional Chinese medicine. METHODS: Fingerprints of 18 Qingkailing injection samples from various manufacturers were obtained by HPLC/ELSD, and quality evaluation was performed by constellation graphical clustering method. RESULTS: HPLC/ELSD and constellation graphical clustering method properly revealed the quality information apparently and accurately. CONCLUSION: It was an apparent, credible and efficient method for quality evaluation of Chinese medicines. PMID- 16342690 TI - [HPLC fingerprint for the Corydalis saxicola Bunting injection]. AB - AIM: To establish the fingerprint of Corydalis saxicola Bunting injection. METHODS: It was performed by HPLC Kromasil column was used with acetonitrile and H2O as mobile phase. The flow rate was 0.5 mL x min(-1). The detection wavelength was 254 nm. The mass spectrometry detection was performed on a trap equipped with an ESI interface and operated in positive-ionization mode. RESULTS: Eleven components of Corydalis saxicola Bunting were identified by LC-MS and 7 of them were quantified. CONCLUSION: The fingerprint chromatogram could represent the characteristics of Corydalis saxicola Bunting injection. Evaluation of resemblances and results of precision were satisfactory. This method could be used as quality and quantity control. PMID- 16342691 TI - [HPLC fingerprint of Fritillaria hupehensis]. AB - AIM: To establish a fingerprint analysis method of Fritillaria hupehensis. METHODS: fingerprint was performed by HPLC-ELSD. Hypersil ODS column was used; the mobile phase was composed of methanol (with 0.05% triethylamine) and water with gradient elution; flow rate was 1.0 mL x min(-1); recording time was 60 min; drift tube temperature was 75 degrees C; gas flow rate was 1.9 L x min(-1). RESULTS: HPLC fingerprint of Fritillaria hupehensis was obtained. CONCLUSION: A reliable method was provided for controlling the quality of Fritillaria hupehensis. PMID- 16342692 TI - [Chemical components in essential oils from tender branches and leaves of Rhododendron]. AB - AIM: To analyze and compare the compositions in essential oils from branches and leaves of Rhododendron simsii Planch. and Rhododendron naamkwanense Merr. METHODS: Essential oils were extracted by water distillation according to Chinese Pharmacopoeia and analyzed by capillary gas chromatography-mass spectrometry as well as chemometrics resolution method and authentic compounds. The relative contents of each component in the essential oils were obtained by normalization of peak areas. RESULTS: A total of 124 components were identified, of which 48 compounds were existed in both of the samples. Ninety four compounds accounted for 84.47% of the essential oil from Rhododendron simsii Planch. and seventy eight components accounted for 90.25% of the total essential oil from Rhododendron naamkwanense Merr. were identified. 72.76% and 88.07% of the components in Rhododendron simsii Planch and Rhododendron naamkwanense Merr., respectively, included oxygen element. They are mainly terpenol, acids and esters. 1-octen-3-ol (4.00%, 7.90%), 1,6-octadien-3-ol, 3,7-dimethyl-(12.60%, 3.48%), 9,12,15-octadecatrienoic acid, [Z, Z, Z]- (1.15%, 45.34%), phytol (15.21%, 8.56%), p-menth-1-en-8-ol (2.15%, 3.29%), and 9,12,15-octadecatrienoic acid, ethyl ester, [Z,Z,Z]- (9.16%, 8.01%) were their common main compounds, which accounted for 44. 27% and 76.58% of the total amount of the two essential oil samples, respectively. In addition, n-hexadecanoic acid (7.73%), 9,12 octadecadienoic acid (1.85%) and tetracosanoic acid, methyl ester (1.38%) were also the main compounds in essential oil from Rhododendron simsii Planch. CONCLUSION: Much higher reliability and accuracy were obtained with the help of chemometrics resolution method and authentic n-alkane standard solutions than those of using GC-MS alone. PMID- 16342694 TI - Mutual information-based degrees of coherence of partially polarized light with Gaussian fluctuations. AB - The spatiotemporal coherence properties of partially polarized light with Gaussian probability distributions are analyzed using the mutual information that is a standard measure of statistical dependence. This approach leads to intrinsic degrees of coherence that have powerful invariance properties and that provide new information in comparison with other recently introduced degrees of coherence. PMID- 16342693 TI - [The influence of absorption enhancer and preparation of paclitaxel on its intestinal mucous membrane permeation detected by enhancement by using electron spin resonance technique]. AB - AIM: To investigate the permeation mechanism of paclitaxel by enhancers and preparation factors. METHODS: The fluidity of mucous membrane and membrane protein conformation changes were determined by using electron spin resonance (ESR) when mucous membrane was treated by several enhancers. At the same time, the factors of penetration of lower dissolution drug across the intestinal mucous membrane were studied in three formulas inclusion complex, microemulsion and injection. RESULTS: Polyethylene glycol (PEG) 1500, hydroxypropyl-beta cyclodextrin (HP-beta-CD) and phospholipid as enhancers could reinforce the permeation of paclitaxle because of loosening of protein conformation in intestinal mucous membrane. Paclitaxel-HP-beta-CD inclusion complex and paclitaxel microemulsion as vehicle could significantly increased permeation kinetic rate of paclitaxel with fluid diffuse method. CONCLUSION: Characteristics of enhancing intestinal absorption of poor dissolution drug had been provided with enhancer the change of membrane fluid. PMID- 16342695 TI - Response of a SiC photodiode to extreme ultraviolet through visible radiation. AB - The responsivity of a type 6H-SiC photodiode in the 1.5-400 nm wavelength range was measured using synchrotron radiation. The responsivity was 0.20 A/W at 270 nm and was less than 0.10 A/W in the extreme ultraviolet (EUV) region. The responsivity was calculated using a proven optical model that accounted for the reflection and absorption of the incident radiation and the variation of the charge collection efficiency (CCE) with depth into the device. The CCE was determined from the responsivity measured in the 200-400 nm wavelength range. By use of this CCE and the effective pair creation energy (7.2 eV) determined from x ray absorption measurements, the EUV responsivity was accurately modeled with no free parameters. The measured visible-light sensitivity, although low compared with that of a silicon photodiode, was surprisingly high for this wide bandgap semiconductor. PMID- 16342696 TI - Brillouin gain analysis for fibers with different refractive indices. AB - We studied an anomalous result in the measured stimulated Brillouin scattering threshold for single-mode optical fibers with different refractive index profiles and effective areas. It was found that, in addition to the optical effective area, the refractive-index-dependent interaction between the optical and the acoustic fields must be considered in order to account for the differences in the stimulated Brillouin scattering threshold between fiber types. A detailed analytical formalism describing the acousto-optic interaction was derived to accommodate the influence of the index profile design on Brillouin scattering. PMID- 16342697 TI - Apodization of elliptical-core two-mode fiber acousto-optic filter based on acoustic polarization control. AB - We propose and demonstrate a novel apodization technique for an elliptical-core two-mode fiber acousto-optic filter by twisting the fiber and adjusting the acoustic polarization. The sidelobe suppression in the filter spectrum was improved from -9.7 to -15.5 dB. A theoretical analysis that includes acoustic birefringence explains the experimental results with good agreement. PMID- 16342698 TI - Strain and temperature sensitivity of a single-mode polymer optical fiber. AB - We have measured the optical phase sensitivity of fiber based on poly(methyl methacrylate) under near-single-mode conditions at 632.8 nm wavelength. The elongation sensitivity is 131 +/- 3 x 10(5) rad m(-1) and the temperature sensitivity is -212 +/- 26 rad m(-1) K(-1). These values are somewhat larger than those for silica fiber and are consistent with the values expected on the basis of the bulk polymer properties. PMID- 16342699 TI - Extended blue supercontinuum generation in cascaded holey fibers. AB - By combining multiple photonic crystal fibers with sequentially decreasing zero dispersion wavelengths we have produced a 1.2 W average-power white-light continuum, covering the visible-near-infrared spectrum from 0.44 to 1.89 microm (10 dB width), with an all-fiber picosecond ytterbium pump laser. Wavelengths as short as the ultraviolet (0.35 microm), and spectral power densities of more than 2 mW/nm in the blue spectral region, have been generated. The process is understood in terms of optimizing four-wave mixing phase matching to enhance short-wavelength generation. PMID- 16342700 TI - Single-frame multichannel blind deconvolution by nonnegative matrix factorization with sparseness constraints. AB - Single-frame multichannel blind deconvolution is formulated by applying a bank of Gabor filters to a blurred image. The key observation is that spatially oriented Gabor filters produce sparse images and that a multichannel version of the observed image can be represented as a product of an unknown nonnegative sparse mixing vector and an unknown nonnegative source image. Therefore a blind deconvolution problem is formulated as a nonnegative matrix factorization problem with a sparseness constraint. No a priori knowledge about the blurring kernel or the original image is required. The good experimental results demonstrate the viability of the proposed concept. PMID- 16342701 TI - Precise subnanometer control of the position of a macro object by light pressure. AB - For the first time to the authors' knowledge, efficient control of the position of a macro object by coherent light was demonstrated. The minimal controllable mechanical displacement induced by the light pressure was 9 pm. No dependence of light pressure on wavelength in a broad wavelength range (405-1560 nm) was observed, as predicted by Maxwell's theory. PMID- 16342702 TI - Combination of a continuous-wave optical parametric oscillator and a femtosecond frequency comb for optical frequency metrology. AB - We combined a tunable continuous-wave optical parametric oscillator and a femtosecond Ti:sapphire laser frequency comb to provide a phase-coherent bridge between the visible and the mid-infrared spectral ranges. As a first demonstration of this new technique we performed a direct frequency comparison between an iodine-stabilized Nd:YAG laser at 1064 nm and an infrared methane optical frequency standard at 3.39 microm. PMID- 16342703 TI - Two-point time-series measurements of minor-species concentrations in a turbulent nonpremixed flame. AB - We report a technique that is capable of making simultaneous two-point time series measurements of minor-species concentrations in turbulent flames. The experimental setup, which incorporates picosecond time-resolved laser-induced fluorescence, has a spatial resolution of less than 250 microm and a temporal resolution of less than 100 micros, which spatially and temporally resolve microscales in many turbulent flows. Two-point time-series data are given for a standard turbulent nonpremixed flame at Re= 10,000, including a discussion of potential implications. PMID- 16342704 TI - Dynamic speckle effect induced by an acousto-optic deflector for fast range sensing. AB - A technique for fast distance measurements based on continuous frequency measurements of the power modulation of spatially filtered scattered light is proposed. For what is to our knowledge the first time, it is shown that the technique works when laser beam scanning is performed with an acousto-optic deflector. The most impressive feature of the proposed technique is that it works at very high scanning speed, providing an extremely fast response time. Experimental verification of the technique is demonstrated at a scanning speed as as high as 130 m/s. The proposed method of range sensing allows the design of a distance sensor possessing a response time as fast as 80 ns. PMID- 16342705 TI - High-refractive-index measurement with an elastomeric grating coupler. AB - An elastomeric grating coupler fabricated by the replica molding technique is used to measure the modal indices of a silicon-on-insulator (SOI) planar waveguide structure. Because of the van der Waals interaction between the grating mold and the waveguide, the elastomeric stamp makes conformal contact with the waveguide surface, inducing a periodic index perturbation at the contact region. The phase of the incident light is changed to match the guided modes of the waveguide. The modal and bulk indices are obtained by measuring the coupling angles. This technique serves to measure the high refractive index with a precision better than 10(-3) and allows the elastomeric stamp to be removed without damaging the surface of the waveguide. PMID- 16342706 TI - Design for beam splitting components employing silicon-on-insulator rib waveguide structures. AB - We present a new design for beam splitting components employing a silicon-on insulator rib waveguide structures. In the new design, a high-index thin-film layer is deposited in the rib section to reduce the wave field dispersive tails in the slab section and accordingly render the mode field a confined spot. This in turn improves the beam splitting performance of some conventional waveguide components such as y branches and multimode interference couplers (MMICs), in terms of the excess loss, fiber coupling loss, and compactness of these components. For a 1 x 2 y-branch beam splitter, the excess loss can be as small as 0.43 dB in the new design, which is much lower than that for a conventional rib waveguide structure (which is 1.28 dB). For a 1 x 2 MMIC in our example, the new rib waveguide structure presents an excess loss of 0.064 dB for the TE mode and 0.046 dB for the TM mode, with negligible nonuniformity in dimensions of 30 microm x 1040 microm, whereas its counterpart (i.e., the one with the same dimensions but without a thin-film layer) presents an excess loss of approximately 0.86 dB for both modes. A conventional MMIC must have dimensions larger than 70 microm x 5650 microm to maintain almost the same low excess loss. PMID- 16342707 TI - Compensating intermodal dispersion in photonic crystal directional couplers. AB - Intermodal dispersion between the supermodes of a directional coupler may induce undesirable pulse breakup in a sufficiently large device. When this happens the device will no longer exchange power between its arms, and the extinction ratio is completely canceled. It is shown that, by carefully designing the coupling area of the directional coupler, one may compensate for intermodal dispersion. The compensating device should accomplish three basic requirements: inverse intermodal dispersion, balanced coupling of each supermode, and maximum power transfer while preserving the sign of the slope of the coupling coefficient with frequency for multiplexing-demultiplexing applications. This structure is designed and optimized with a genetic algorithm. PMID- 16342708 TI - Continuous-wave laser operation of Yb:LuVO4. AB - We report on the room-temperature spectroscopic properties and continuous-wave laser operation of a new Yb:LuVO4 crystal. The peak absorption cross section for the pi-polarization is 8.42 x 10(-20) cm2 at 985 nm, and the stimulated emission cross section at 1020 nm is 1.03 x 10(-20) cm2. An output power of 0.36 W at 1041 nm was obtained with a slope efficiency of 47% by use of Ti:sapphire laser pumping. With diode pumping the output power reached 1.05 W at the highest available incident pump power of 6.8 W. PMID- 16342709 TI - Ultrafast deflection of spatial solitons in AlxGa(1-x)As slab waveguides. AB - We demonstrate ultrafast all-optical deflection of spatial solitons in an AlxGa(1 x)As slab waveguide, using 190 fs, 1550 nm pulses to generate and deflect the spatial soliton. The steering beam is focused onto the top of the waveguide near the soliton pathway, and the soliton is steered by refractive-index changes induced by optical Kerr, or free-carrier (Drude), effects. Angular deflections up to 8 mrad are observed. PMID- 16342710 TI - Upper threshold for stability of multipole-mode solitons in nonlocal nonlinear media. AB - We address the stability of multipole-mode solitons in nonlocal Kerr-type nonlinear media. Such solitons comprise several out-of-phase peaks packed together by the forces acting between them. We discover that dipole-, triple-, and quadrupole-mode solitons can be made stable, whereas all higher-order soliton bound states are unstable. PMID- 16342711 TI - Incoherent blocker soliton interactions in Kerr waveguide arrays. AB - We have observed the incoherent interaction between a highly confined (blocker) soliton and wide, moving signal beams of a different wavelength in a one dimensional discrete Kerr medium. Digital switching of the blocker solitons to successive adjacent channels was measured with increasing signal power via both one and two cascaded interactions in an AlGaAs waveguide array, operations equivalent to a reconfigurable three-output router. PMID- 16342712 TI - Incoherent light-induced self-organization of molecules. AB - Although coherent light is usually required for the self-organization of regular spatial patterns from optical beams, we show that peculiar light-matter interaction can break this evidence. In the traditional method of recording laser induced periodic surface structures, a light intensity distribution is produced at the surface of a polymer film by an interference between two coherent optical beams. We report on the self-organization followed by propagation of a surface relief pattern. It is induced in a polymer film by using a low-power and small size coherent beam assisted by a high-power and large-size incoherent and unpolarized beam. We demonstrate that we can obtain large size and well-organized patterns starting from a dissipative interaction. Our experiments open new directions to improving optical processing systems. PMID- 16342713 TI - Fabrication of three-dimensional structures by three-photon polymerization. AB - We report the fabrication of three-dimensional structures of submicrometer resolution by three-photon polymerization. This resolution has been achieved by polymerizing ORMOCER, a UV photocurable organic-inorganic hybrid material, with an ultrafast laser irradiation at 1028 nm. To our knowledge, this is the first demonstration of three-photon polymerization, a process that may allow the fabrication of components of very high resolution. PMID- 16342714 TI - Linear-to-circular polarization transformation upon optical tunneling through an embedded low-index film. AB - Quarter-wave retardation can be achieved in optical tunneling through a low-index thin film that is surrounded by a medium of higher refractive index, for any index ratio N>2.414, over a range of incidence angles that is determined by N. A novel frustrated-total-internal-reflection circular polarization beam splitter (CPBS) is proposed that uses an adjustable air gap between two Ge prisms. This tunable CPBS operates over a broad (2-12 microm) IR spectral range by controlling the air-gap thickness to match one-tenth of the wavelength of light by use of a piezoelectric transducer. PMID- 16342715 TI - Terbium-based extreme ultraviolet multilayers. AB - We have fabricated periodic multilayers that comprise either Si/Tb or SiC/Tb bilayers, designed to operate as narrowband reflective coatings near 60 nm wavelength in the extreme ultraviolet (EUV). We find peak reflectance values in excess of 20% near normal incidence. The spectral bandpass of the best Si/Tb multilayer was measured to be 6.5 nm full width at half-maximum (FWHM), while SiC/Tb multilayers have a more broad response, of order 9.4 nm FWHM. Transmission electron microscopy analysis of Si/Tb multilayers reveals polycrystalline Tb layers, amorphous Si layers, and relatively large asymmetric amorphous interlayers. Thermal annealing experiments indicate excellent stability to 100 degrees C (1 h) for Si/Tb. These new multilayer coatings have the potential for use in normal incidence instrumentation in a region of the EUV where efficient narrowband multilayers have not been available until now. In particular, reflective Si/Tb multilayers can be used for solar physics applications where the coatings can be tuned to important emission lines such as O V near 63.0 nm and Mg X near 61.0 nm. PMID- 16342716 TI - Subfemtomole detection of small molecules with microsphere sensors. AB - We investigated the feasibility of using a silica microsphere sensor for detection of small molecules. Using the silica molecules (60 Da) at the sphere's surface as a model system, we measured the spectral shifts in the whispering gallery modes (WGMs) when the sphere size was decreased by a hydrofluoric acid (HF) solution. The results demonstrate that our sensor is capable of detecting a 4 pm (or 0.01 layer of silica) decrease in sphere radius, corresponding to a change of 0.4 fmol silica molecule. These results suggest that small molecules can be detected in trace quantities at the surface of an optical microsphere sensor. PMID- 16342717 TI - Disorder-immune confinement of light in photonic-crystal cavities. AB - We demonstrate by finite-difference time-domain simulations in 2D and 3D that optical cavities in realistic finite photonic crystals have lifetimes and modal volumes that are essentially insensitive to disorder (of various types, including surface disorder and randomized positions), even with unphysically large disorder. A lifetime Q = 10(8) is demonstrated in a 3D single-mode cavity with a half-wavelength mode diameter using only eight vertical periods of a disordered crystal. PMID- 16342718 TI - Thermal image encryption obtained with a SiO2 space-variant subwavelength grating supporting surface phonon-polaritons. AB - Space-variant partially polarized thermal emission is investigated. We show that by coupling surface phonon-polaritons to a propagating field, large anisotropy of the emissivity is obtained within a narrow spectral range. We experimentally demonstrate this effect by fabricating a space-variant subwavelength grating on a SiO2 substrate to encrypt an image in the polarization state of a thermal radiation field. PMID- 16342719 TI - Cut-wire pairs and plate pairs as magnetic atoms for optical metamaterials. AB - We study the optical properties of metamaterials made from cut-wire pairs or plate pairs. We obtain a more pronounced optical response for arrays of plate pairs, a geometry that also eliminates the undesirable polarization anisotropy of the cut-wire pairs. The measured optical spectra agree with simulations, revealing negative magnetic permeability in the range of telecommunications wavelengths. Thus nanoscopic plate pairs might serve as an alternative to the established split-ring resonator design. PMID- 16342720 TI - Optical anisotropy in single-walled carbon nanotubes. AB - Optical anistropy at optical communication wavelength was observed in films of vertically aligned single-walled carbon nanotubes (SWNTs). We report the control of both the polarization state and transmission of incoming light at 1550 nm by azimuthal and axial tilting of SWNT film about its aligned axis. The experiments reveal that the polarization state of light is susceptible to the azimuthal angle of the aligned direction of a SWNT having semiconductor characteristics and the intensity of the output beam after SWNT film shows cosine function dependence on the axial tilting angle. PMID- 16342721 TI - Azimuthal prism effect with partially blocked vortex-producing lenses. AB - We report a new effect in which the diffraction from a partially blocked angular phase pattern occupying half of the input plane produces a partial vortex output pattern that is rotated by 90 degrees compared with the input. The energy is sent into a different quadrant of the output plane from the input plane. The rotation direction depends on whether the angular phase pattern is clockwise or counterclockwise. When we combine clockwise and counterclockwise angular phase patterns on separate horizontal halves of the input plane, we create an interference effect in one half of the output plane, while the other half remains dark. PMID- 16342722 TI - Low-light-level photon switching by quantum interference. AB - We report an experimental demonstration of low-light-level photon switching by quantum interference in laser-cooled 87Rb atoms. A resonant probe pulse with an energy per unit area of one photon per lambda2/2pi propagates through the optically thick atoms. Its energy transmittance is greater than 63%, or a loss of less than e(-1), because of the effect of electromagnetically induced transparency. In the presence of a switching pulse with an energy per unit area of 1.4 photons per lambda2/2pi, the energy transmittance of the same probe pulse becomes less than 37%, or e(-1). This substantial reduction of probe transmittance caused by switching photons may lead to potential applications in single-photon-level nonlinear optics and manipulation of quantum information. PMID- 16342723 TI - Spontaneous emission in the near field of two-dimensional photonic crystals. AB - We show theoretically that photonic crystal membranes cause large variations in the spontaneous emission rate of dipole emitters, not only inside but also in the near field above the membranes. Our three-dimensional finite-difference time domain calculations reveal an inhibition of more than five times and an enhancement of more than ten times for the spontaneous emission rate of emitters with select dipole orientations and frequencies. Furthermore, we demonstrate theoretically the potential of a nanoscopic emitter attached to the end of a glass fiber tip as a local probe for mapping the large spatial variations of the photonic crystal local radiative density of states. This arrangement is promising for on-command modification of the coupling between an emitter and the photonic crystal in quantum optical experiments. PMID- 16342724 TI - Rabi oscillation damped by exciton leakage and Auger capture in quantum dots. AB - The decoherence of Rabi oscillation (RO) caused by biexciton, population leakage to the wetting layer (WL), and Auger capture in semiconductor quantum dots is theoretically analyzed with multilevel optical Bloch equations. The corresponding effects on the quality factor of RO are also discussed. We have found that the biexciton effect is relatively trifling, as the pulse duration is longer than 5 ps. The population leakage to the WL leads to a decrease of the RO average even though the damping rate is similar to that observed in the experiment. Auger capture in quantum dots results in RO damping that is consistent with the experimental data, which implies that Auger capture is an important decoherence process in quantum dots. PMID- 16342725 TI - Experimental observation of universality in depolarized light scattering. AB - Experimental results on light depolarization due to multimode scattering are reported. By means of polarization tomography, we characterize the depolarizing power and the polarization entropy of a broad class of optically scattering media and confirm the recently predicted universal behavior of these two quantities [Phys. Rev. Lett. 94, 090406 (2005)]. PMID- 16342726 TI - Velocity measurements by cavity ringdown spectroscopy. AB - We demonstrate velocity measurements of gas-phase particles by using cavity ringdown spectroscopy (CRDS). Velocity information is inferred from the Doppler shift contributions to the measured absorption line shape. Because in CRDS the laser beam propagates back and forth within the optical cavity, a measured absorption feature is both upshifted and downshifted; i.e., it is split by the velocity component parallel to the optical axis. The splitting of the absorption features allows direct velocity measurements to be made without requiring an external frequency reference. The CRDS velocity measurement approach is demonstrated for sputtered molybdenum atoms in a low-pressure (collisionless) environment. PMID- 16342727 TI - Broadband coherent anti-Stokes Raman scattering spectroscopy of nitrogen using a picosecond modeless dye laser. AB - Broadband picosecond coherent anti-Stokes Raman scattering (CARS) spectroscopy of nitrogen is demonstrated using 145-ps pump and probe beams and a 115-ps Stokes beam with a spectral bandwidth of 5 nm. This is, to our knowledge, the first demonstration of broadband CARS using subnanosecond lasers. The short temporal envelope of the laser pulses and the broadband spectral nature of the Stokes beam will enable nonresonant-background-free, single-shot, or time-dependent spectroscopy in high-pressure or hydrocarbon-rich environments. Successful correlation of room-temperature broadband picosecond N2 CARS with a theoretical spectrum is presented. PMID- 16342728 TI - High-energy (nanojoule) femtosecond pulse delivery with record dispersion higher order mode fiber. AB - Delivery of high peak-power femtosecond pulses with fibers is constrained by nonlinear distortions accumulated during pulse propagation. We address this problem with a novel, to our knowledge, fiber schematic, where the pulse propagates in a small Aeff (18 microm2) but highly dispersive (record value of approximately -900 ps/nm km) medium, enabled by transmission in the LP02 mode of a few-mode fiber. The novel fiber yields a low dispersion-to-nonlinear-length ratio (due to its large dispersion) despite its small Aeff, hence enabling mitigation of nonlinearities. This enables fiber delivery of distortion-free <150 fs, approximately 1 nJ, and 840 nm pulses--an order-of-magnitude improvement over single-mode fibers of similar Aeff. PMID- 16342729 TI - Reconfigurable generation of high-repetition-rate optical pulse sequences based on time-domain phase-only filtering. AB - We propose and demonstrate a fiber-based phase-only filtering technique for programmable optical pulse shaping, in which the filtering operation is implemented in the time domain by means of an electro-optical (EO) phase modulator. The technique has been applied for generating customized ultrahigh repetition-rate optical pulse sequences (>40 GHz) from single input pulses by driving the EO phase modulator with a periodic electronic waveform (RF tone). The generated output pulses are replicas of the input pulse and both the repetition rate and the envelope profile of the generated sequences can be controlled and tuned electronically using this approach. PMID- 16342730 TI - Generation of phase-locked and tunable continuous-wave radiation in the terahertz regime. AB - Broadly tunable phase-stable single-frequency terahertz radiation is generated with an optical heterodyne photomixer. The photomixer is excited by two near infrared CW diode lasers that are phase locked to the stabilized optical frequency comb of a femtosecond titanium:sapphire laser. The terahertz radiation emitted by the photomixer is downconverted into RF frequencies with a waveguide harmonic mixer and measurement-limited linewidths at the Hertz level are demonstrated. PMID- 16342731 TI - Yb3+-doped YVO4 crystal for efficient Kerr-lens mode locking in solid-state lasers. AB - We report the first demonstration, to our knowledge, of soft-aperture Kerr-lens mode locking in a diode-pumped femtosecond Yb3+:YVO4 laser. Near-transform limited pulses as short as 61 fs are generated around a center wavelength of 1050 nm with an output power of 54 mW and a pulse repetition frequency of 104.5 MHz. This is, to our knowledge, the shortest pulse generated directly from an Yb laser having a crystalline host material. The femtosecond operation has a mode-locking threshold at an absorbed pump power of 190 mW. The nonlinear refractive indexes of the Yb3+:YVO4 crystal have been measured to be 19 x 10(-16) cm2/W and 15 x 10( 16) cm2/W for the sigma and pi polarizations, respectively, at 1080 nm. PMID- 16342732 TI - Characterization of nonlinear phase shifts by use of the temporal transport-of intensity equation. AB - A temporal transport-of-intensity equation is derived to describe the evolution of the instantaneous power of the electric field in a dispersive medium. As its spatial counterpart, which can be used to measure wavefronts using the intensity measured after various amounts of diffraction, the temporal transport-of intensity equation allows the determination of temporal phase shifts using only intensity measurements in a direct noninterferometric way. The application to the characterization of nonlinear phase shifts induced by self-phase modulation and cross-phase modulation is used to determine the nonlinear coefficient of a highly nonlinear fiber. PMID- 16342733 TI - Evaluation by chemical parameters of the pollution state of the agricultural industrial settlement of the Fucino Plain. AB - In the present work are reported the results of a monitoring on a vast scale, carried out through evaluation of opportune chemical parameters, of the pollution state of the agricultural-industrial settlement of the Fucino Plain. The parameters took into consideration have been the presence of wide consumption pesticides and of ionic species as Cl-, NO2-, NO3-, NH4+, the quantification of the dissolved oxygen, of the temperature, of the conductivity, of pH and eH. Collected data are used for chemometric elaboration. The water systems examined, by means of drawing campaigns carried out at regular intervals in winter, spring, summer, and autumnal seasons, are represented by superficial waters constituting a network of irrigation canals fed by stratum and meteoric waters. In this work are reported the results relative to the drawing campaigns. PMID- 16342734 TI - Evaluation of chlorinated by-products in drinking waters of Central Friuli (Italy). AB - Drinkable water supplied by aqueducts undergoes preliminar potabilization which, in Italy, is mainly accomplished by chlorine addition. The bactericidal action involved in this process is always accompanied by chlorination and oxidation of organic species (mainly humic and fulvic acids) naturally present in treated waters, so that many disinfection by-products (DBPs) are formed, such as trihalomethanes (THMs) and halo-acetic acids (HAA), which can represent a chemical risk for public health. The aim of this study was the monitoring of DBPs in drinking water disinfected by chlorination, supplied by four different aqueducts of Central Friuli (Italy). DBP evaluations were performed in water samples consisting of both input and output of disinfection plants. The results of analytical determinations were worked out to provide the THM and HAA parameters for disinfected waters, while in feeding waters the following different conventional parameters were adopted: (i) trihalomethanes formation potential (THMFP), (ii) halo-acetic acids formation potential (HAAFP) and (iii) UV absorbance at 254 nm (UV254). The quite moderate content of chlorinated products found in all samples considered highlighted the excellent quality of potabilized waters available in Central Friuli. Moreover, our results confirmed that the majority of DBPs formed when chlorine is used for water disinfection consists of THMs, while chlorites and chlorates prevailed when potabilization is accomplished by using chlorine dioxide. Finally, simple UV254 monitoring turned out to be a profitable approach for the determination of chlorinated by-products only when THMs prevail among DBPs. PMID- 16342735 TI - Determination of polycyclic aromatic hydrocarbons (PAHs) and total fats in human milk. AB - A sensitive and reliable analytical method was developed for the simultaneous extraction of PAHs and total fats and their determination in the human milk. The method involved a liquid-liquid extraction of PAHs and fats, followed by the gravimetric determination of the latter. PAHs were separated from lipids by size exclusion chromatography eluting with methylene chloride and analysed by gaschromatography coupled with mass spectrometry (GC/MS). The recovery of analytes was in the range of 42-101% and agreed well with their boiling temperatures (R2=0.779). Precision of the method was found between 7.6 and 19%. Quantification and detection limits for individual PAHs ranged from 0.011 to 0.032 and from 0.006 to 0.022 microg/Kg milk (wet weight), respectively. Quantification limit for the total fat determination was 0.26 g/Kg milk (wet weight). This procedure, applied to milk samples of ten healthy, non-smoking, Italian primiparae, living in rural or low-traffic zones, allowed for the identification and quantitative determination of naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo(a)anthracene, benzo(b)fluoranthene and benzo(k)fluoranthene in the mean concentration range 0.114-6.95 microg/Kg milk (wet weight). The most volatile compounds, which were not investigated elsewhere, were found at much higher concentrations than those observed for the others. No relation was found between PAH and total fat concentrations. PMID- 16342736 TI - The protonation of polyacrylate in seawater. Analysis of concentration effects. AB - The protonation of polyacrylate (PAA, MW 2 kDa) was studied potentiometrically at 25 degrees C, in mixed electrolyte aqueous solution simulating the composition of seawater, in the salinity range 30 < or = S < or = 40. The salt composition of different solutions was varied in order to study its effect on apparent protonation constants. Results were analysed using two fairly different approaches: by simple regression analysis on a combination of concentration parameters and salinity, and by canonical correlation analysis. Unexpectedly we found that variations on protonation constants, due to the different relative component concentrations, are fairly low, revealing a sort of buffering capacity of seawater respect to protonation properties of polyacrylate (and likely for other HMW and LMW polycarboxylates). The intrinsic protonation constant of polyacrylate 2 kDa at 25 degrees C and 35 salinity is log K(H*) = log K(int) = 4.399 +/- 0.004. The use of different pH scales and standard seawater compositions is also discussed. PMID- 16342737 TI - Use of multivariate analysis of MIR spectra to study bread staling. AB - Different kinds of bread, stored at constant temperature and at controlled humidity conditions for a week since their manufacturing date, were analysed by Attenuated Total Reflectance-Fourier Transform InfraRed (ATR-FTIR) spectroscopy. The collected spectra were processed by Principal Component Analysis (PCA), in order to evaluate the changes occurring during bread ageing. For the sake of comparison, the 1060-950 cm spectral window has been also investigated by curve fitting methods. It was observed that the first PC increases monotonically with ageing of samples. Furthermore, the more influential variables on PC1 correspond to spectral regions where are located stretching and bending bands, which are mainly attributed to typical starch bonds vibrations. PMID- 16342738 TI - Distribution of heavy metal concentrations in surface sediments in Dubai Creeks, United Arab Emirates. AB - Dubai is developing rapidly and many developmental activities are concentrated around its Creek. The present study reports the lateral distribution of heavy metals and compares it with local historical record of heavy metal concentrations. For this purpose surface sediment samples were collected and analyzed for metal contents, total organic carbon content (TOC), mineralogy and grain size. The percentages of the different grain size fraction of the collected sediments were as follow 65% for sand size, 15% for silt size fraction, and the rest accounted for clay size fraction. The microscopic analyses indicate that the sediment composed mainly from carbonate and quartz with traces of rock fragments. Such mineral composition is not believed to be a potential source of heavy metal. The study found that the average recorded heavy metal concentrations in the collected sediment samples were 87, 96, 127, 38.5, and 279 ppm for Cr, Cu, Ni, Pb, and Zn, respectively. Those values were slightly higher than metal concentrations recoded in 2001 with 1.22 (Cr), 2.5 (Cu), 2.87 (Ni), 0.69 (Pb), and 2.1 (Zn) folds. However, in 2001 and 2003 the measured metal contents, along the creek, were lower than those of the average earth crust. Along the Creek most metals recorded the highest concentrations in the upper reach of the Creek. The distribution of the measured heavy metals was not affected significantly with the TOC values. The present study also documented obvious related point sources of pollution. PMID- 16342739 TI - Synthesis and efficiency of a chelating fiber for preconcentration and separation of trace Au(III) and Pd(IV) from solution samples. AB - A novel poly(acryl-benzoylamidrazone-acryl-benzoylhydrazine) chelating fiber was synthesized simply from nitrilon (polyacrylonitrile fiber). The structure of the chelating fiber was verified with the help of FT-IR spectrum. The parameters influencing the efficiency of the fiber for concentrating trace amounts of Au(III) and Pd(IV) were investigated in detail. Trace Au(III) and Pd(IV) were enriched and separated from real sample solutions and detected using inductively coupled plasma atomic emission spectrometry (ICP-AES) with satisfactory results. The experiments show that the method is rapid, precise, simple and convenient to use. PMID- 16342740 TI - Electrochemical determination of 6-mercaptopurine at silver microdisk electrodes. AB - The electrochemical behaviour of 6-mercaptopurine (6-MP) at a microdisk electrode is investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The results indicate that 6-MP can be strongly adsorbed on the surface of the static mercury drop electrode (SMDE) and reacts with Ag+ ions which are produced at positive potentials. 6-MP yields a well-defined cathodic stripping signal during the negative scan at about -0.812 V (vs. SCE) in pH 9.0 phosphate buffer solution. The electrode has hence been used for the determination of 6-MP by differential pulse voltammetry (DPV). The linear range is between 2.0x10(-7) and 5.0x10(-5) mol/l, with the calculated detection limit (S/N=3) of 8.0x10(-8) mol/l. The relative standard deviation is 3.0% for eight successive determinations of 4.0x10(-5) mol/l 6-MP. The determination of 6-MP in tablets has also been carried out and satisfactory results have been obtained. PMID- 16342741 TI - Preparation of poly(propyleneimine) dendrimer immobilized silica gel and its application as novel microcolumn packing for the on-line FI preconcentration and separation of platinum. AB - A G4.0 poly(propyleneimine) dendrimer immobilized silica gel (PPID-SG) was prepared with a divergent approach from gamma-aminopropyl silica gel (APSG) by repeating the reactions: 1) cyanoethylation of -NH2 and 2) reduction of -CN. Then it was successfully applied as the microcolumn packing for the on-line flow injection (FI) preconcentration and separation and flame atomic absorption spectrometry (FAAS) determination of Pt. A limit of detection (LOD) of 0.014 microg/mL was obtained when 0.5 microg/mL Pt in chloride acid of PH = 3 was preconcentrated with a sample flow rate of 4.6 mL/min for 60s and the relative standard deviation (RSD) was 2.65%. Coexisting metal ions with concentration of 5.0 mg/mL and anion ions of 20.0 mg/ml have no interference in the determination of Pt. The proposed method was successfully applied to the determination of Pt in nickel alloy and anode slime samples. PMID- 16342742 TI - Synthesis of Ta2O5 nanorods in the presence of poly L-lysine by a sol-gel process. PMID- 16342743 TI - Precolumn derivatization liquid chromatography with mass spectrometry assay for the determination of glucosamine in small volume human plasma. PMID- 16342744 TI - Optimized conditions for the solar photocatalytic degradation of bisphenol A in water using zinc oxide. PMID- 16342745 TI - Water hyacinth (Eichornia crassipes) as a natural tool for pollution control. PMID- 16342746 TI - [Networks of care, networks of health: did you say "networks"?]. PMID- 16342747 TI - [Networks, legal and practical aspects]. PMID- 16342748 TI - [Health networks, theory and practice]. PMID- 16342749 TI - [REDIAB, story of a network]. PMID- 16342750 TI - [Oncology in the Loire, a battle against cancer]. PMID- 16342751 TI - [Partial sequence homology of FtsZ in phylogenetics analysis of lactic acid bacteria]. AB - FtsZ is a structurally conserved protein, which is universal among the prokaryotes. It plays a key role in prokaryote cell division. A partial fragment of the ftsZ gene about 800bp in length was amplified and sequenced and a partial FtsZ protein phylogenetic tree for the lactic acid bacteria was constructed. By comparing the FtsZ phylogenetic tree with the 16S rDNA tree, it was shown that the two trees were similar in topology. Both trees revealed that Pediococcus spp. were closely related with L. casei group of Lactobacillus spp. , but less related with other lactic acid cocci such as Enterococcus and Streptococcus. The results also showed that the discriminative power of FtsZ was higher than that of 16S rDNA for either inter-species or inter-genus and could be a very useful tool in species identification of lactic acid bacteria. PMID- 16342752 TI - [Identification and phylogenetic study of pathogenic bacteria causing ulcer disease of cultured Turbot (Scophthalmus maximus)]. AB - An outbreak of ulcer disease of the cultured turbot Scophthalmus maximus L. occurred in an indoor farm in Haiyang, Shandong Province in the sweltering August of 2004 and caused significant mortality. At the beginning, infected turbots displayed sluggish swimming and anorexia. Several days later, the turbot eyes swelled, the fins and tail turned red, and the back gradually ulcerated. Dissection of the moribund fish showed that the ulcerative gills paled, the liver becamed bloodshot, the kidney and gallbladder swelled, the intestinal wall became filmy and bloodshot. The time course from appearance of disease signs to death lasted about a week. A dominant strain of bacteria, which was Gram-negative and short rod with single polar flagellum under electron microscope, was isolated from the diseased turbot and designated as H1. In artificial infection test, all fish of the experimental groups died in a week after intramuscularly injected with bacterial suspension, while the control group showed no signs in 10 d post challenge. The moribund experimental fish had similar gross signs as the natural infected fish. The bacteria re-isolated from the challenged fish also had the same characteristics as H1, which proved that the isolate H1 was the pathogenic bacteria that triggered this ulcer disease. Different methods were used to identify the pathogenic bacteria. The identification result by API 20NE and API 20E system indicated that H1 was Aeromonas hydrophila, with 99% reliability and 61.5% respectively. While traditional biochemical identification revealed that H1 exhibited relatedness to Vibrio harveyi. In order to confirm the different result, a 1424 bp sequence of H1's 16S rDNA was amplified and compared with other Vibrio spp. in GenBank, homology analysis and phylogenetic study showed that H1 has the highest similarity to V. harveyi, with 99% identity. According to morphological features, physiological and biological characteristics and 16S rDNA homology comparison of the bacteria, the pathogenic bacteria were V. harveyi. Drugs sensitivity test showed that the pathogenic bacteria were highly sensitive to nitrofurantoinum and ceftriaxone sodium etc. This is the first report that V. harveyi was found as the pathogenic bacteria of cultured turbot in China. The research suggests that V. harveyi should be regarded as an important pathogen of turbot and can causes ulcer disease under conditions of high temperatures. Therefore, it is necessary to prevent against ulcer disease in culture turbot in summer. PMID- 16342753 TI - [Bacterial 16S rDNA sequence analysis of Siberian tiger faecal flora]. AB - Bacterial 16S rDNA library of Siberian tiger was developed and 15 different clones were obtained using EcoR I and Hind III in restriction fragment length polymorphism analysis. DNA sequencing and similarity analysis showed that 10 clones matched corresponding Clostridium sequences, of which 6 sequences had over 99% similarity with Clostridium novyi type A, and 4 sequences had 97% similarity with Swine manure bacterium RT-18B, which identified as Peptostreptococcus spp. The other five 16S rDNA sequences had 94% - 95% similarity with Clostridium pascui, Clostridium tetani E88, Clostridium sp. 14505 Clostridium perfringens and Carnobacterium sp. R-7279 respectively. PMID- 16342754 TI - [Construction of an engineered Acidithiobacillus caldus with high-efficiency arsenic resistance]. AB - Using the recombinant technique in vitro, a new arsenic resistance plasmid pSDRA4 was constructed by subcloning the arsenic resistance genes from plasmid pUM3 into the wide-host-range IncQ plasmid pMMB24 with the hybrid trp-lac ( tac ) promoter, and followed by deleting the regulative gene of the promoter, the lacIQ gene. Then plasmid pSDRA4 was introduced from E. coli into extremely acidophilic obligately chemolithotrophic Acidithiobacillus caldus by conjugative transfer with a frequency of( 1.444 +/- 0.797) x 10(-4), and the engineered strain of Acidithiobacillus caldus (pSDRA4) for biomining was constructed. The successful transfer demonstrates the development of a conjugational system between strains of E. coli and A. caldus. The recombinant plasmid pSDRA4 is stable in A. caldus. Compared with wild type A. caldus, the level of the arsenic resistance of A. caldus (pSDRA4) is greatly raised from 10mmol/L to 45mmol/L. PMID- 16342755 TI - [Identification of genomic differences in avian pathogenic Escherichia coli using suppression subtractive hybridization analysis]. AB - To identify unique DNA fragments associated with avian pathogenic Escherichia coli strains, suppression subtractive hybridization (SSH) was used. The genome of nonpathogenic E. coli K-12 strain MG1655 was subtracted from the genome of avian highly pathogenic strain E037 (serotype O78) resulting in the identification of 17 specific fragments. And the genome of avian low pathogenic E. coli strain E526 (serotype O2) was subtracted from the genome of avian highly pathogenic strain E058 (serotype O2) resulting in the identification of 32 specific fragments. Sequence homology analysis was done and four types of fragments were identified: plasmid sequences, phage sequences, sequences with known function and sequences without any currently known function. And 12 specific fragments that were not found in E. coli K-12 were identified from two avian E. coli strains. The results suggested that there were some genetic differences between the highly pathogenic strains and low pathogenic or nonpathogenic strains. PMID- 16342756 TI - [Expression of recombinant Vp6 gene of porcine rotavirus A with non-antibiotic Lactobacillus vector in Escherichia coli]. AB - The antigenic determinants of Vp6 gene of porcine rotavirus A was amplified from infected MA 104 cell by the reverse transcription-polymerase chain reaction (RT PCR), the product of which was a 1194bp cDNA segment. Using T-A cloning technique, the PCR product was cloned into pGEM-T vector. Cloning plasmid pGEM-T Vp6 and the prokaryotic shuttle expression vector pW425t between E. coli and Lactobacillus, were digested by SacI and KpnI double enzymes, respectively. The purified Vp6 gene was subcloned into the expression vector pW425t. Thus, the recombinant pW425t-Vp6 was constructed, which then was transformed into the competence thyA gene-mutant E. coli X13. Treated lysates of bacterium were loaded directly onto SDS-PAGE, on which approximately 44.88 kD fusion protein was observed. The protein was further analyzed using Western blot, which indicated that the protein was reactive with the antibody of rotavirus A. The results lay foundation for further studies on the Lactobacillus subunit vaccine and DNA vaccine of Vp6 gene for prevention and control of porcine rotavirus. PMID- 16342757 TI - [Cloning of full-length genes of H5N1 subtype Avian influenza virus strain A/duck/Shandong/093/2004 and analysis of the sequences]. AB - The eight full-length genes, including the 5' and 3' ends of H5N1 subtype Avian influenza virus (A/duck/ Shandong/093/2004) were amplified by using the universal primers and H5 specific primers. The method used for the amplification of Avian influenza virus's full-length sequence was more easily and rapidly than that of rapid amplification of cDNA ends assay (RACE). The amplified segments were cloned into the T vector PCR 2.1, respectively. Three to five positive clones of each gene were sequenced and the same two sequencing results of the full-length genes were obtained. The phylogenetic analysis results showed that all the eight segments of the A/duck/Shandong/093/2004 were different from the A/Quail/Hongkong/G1/97 and A/Chicken/Beijing/1/94, but showed highly similarity (99% and above) to that of four H5N1 strains, which were isolated in 2002 in duck. It revealed that this strain was resulted from re-assortment of H5N1 rather than H9N2. The NA sequence of A/D/SD/04 was analyzed and the result demonstrated that there are 20 amino acids missing in 48 - 68 sites, however, there was no residue lost in NS gene in 263 to 277 sites. The motif of HA cleavage site is PQRERRRKKR/G, which is the characteristic of HPAIV. The 226 amino acid residue was Met (M), which can react with both Aalpha-2, 3Gal and SAalpha-2, 6Gal receptor. And the 627 residue of PB2 was Glutamic acid (E). The result mentioned above confirmed that H5N1 subtype AIV has multiple determinants in its virulence. A/D/SD/04 is the mid-strain evolving from HPAIV to a virulent strain of mammal. PMID- 16342758 TI - [High level expression of recombinant chicken interferon-gamma in insect cells]. AB - The recombinant transfer vector pFastBacl-ChIFN-y was constructed by plasmid pcDNA-ChIFN-gamma digested with EcoR I and Not I enzymes and cloned into pFastbacl. Then the transfer vector was transformed into E. coli competent cells DH10Bac which contained the bacmid with amini-attTn7 target site and the helper plasmid. The recombinant bacmid-ChIFN-gamma was generated by transposing themini Tn7 element located in pFastBacl-ChIFN-gamma to themini-attTn7 attachment site on the Bacmid. Subsequently the recombinant Bacmid-ChIFN-gamma was transfected into the Sf9 insect cells mediated by lipofectin to produce recombinant baculovirus and express recombinant ChIFN-gamma (rChIFN-gamma) products. The result showed that the rChIFN-gamma was successfully expressed in Sf9 cells infected with the recombinant virus by indirect immunofluorescence assay (IFA) at 5 days post transfection. The biological activity of rChIFN-gamma was identified by its inhibition to Vesicular stomatitis virus-induced cytotoxicity of chicken embryonic fibroblasts (CEF) in vitro. The results showed that the most efficient expression of rChIFN-gamma could be obtained at 96h post-infection with multiplicity of infection (MOI) equal to 1. It is interesting that the viruses such as Avian influenza virus H5N1 or Marek's disease virus (GA strain) could not grow in CEF pre-treated with rChIFN-gamma. Cell pathogenic efficient (CPE) in the CEF infected with H5N1 and GA strain is apparently inhibited by the rChIFN-gamma. However only difference between the HA titres of the supernatant of the pre treated cells is observed without any obvious inhibition effect in CEF infected with Newcastle disease virus (F48E8 strain). PMID- 16342759 TI - [Bioactivity of Laetiporus sulphureus var. sulphureus metabolites in liquid culture]. AB - The liquid culture of Laetiporus sulphureus var. sulphureus was lethal against fruit fly. It was found that extracellular metabolites were primary causation of the lethal effect against fruit fly, which was influenced by pH value. Isolation and analysis with ion-exchange resin column chromatography and HPLC demonstrated that oxalic acid was present in supernatant of Laetiporus sulphureus var. sulphureus, and it was one of the contributing factors to lethal effect against fruit fly and decrease of pH value of culture system. When cultured in airlift reactor, concentration of oxalic acid, quantity of mycelia and pH value was correlated with each other. Further analysis on elution revealed that a kind of oligidic pigment of amaranth in alkaline condition were also lethal against fruit fly. PMID- 16342760 TI - [Purification and properties of anitibiotic from Burkholderia cepacia CF-66]. AB - Burkholderia cepacia CF-66 isolated from compost showed antifungal activity against plant pathogens such as Rhizoctomia solani and some other fungi. CF66I produced by Burkholderia cepacia CF-66 was separated by gel chromatography using Sephadex-75pg and Sephacryl S-100 column. CF66I has very high degree of stability of antifungal activity under higher temperature and alkaline conditions. Some organic solvents could improve its activity at low concentration. The research for the structure of CF66I find that it is a compound with amide bonds and its main structural units is (CH2CH2O)n. PMID- 16342761 TI - [Purification and properties of manganese peroxidase from Trametes versicolor]. AB - Two manganese peroxidase (MnP) active fractions D1 and D2 were got from the extracellular culture of Trametes versicolor by using ammonium sulfate precipitation, DEAE-cellulose DE52 chromatography. MnP1 was purified to electrophoretic homogeneity from the D2 by Phenyl Sepharose 6 Fast Flow chromatography and MnP2 was purified to electrophoretic homogeneity from the D1 by Sephacryl S-200HR chromatography and Phenyl Sepharose 6 Fast Flow chromatography. The specific activities of two MnP isozymes are 579.1U/mg and 425.0U/mg; purification folds are 17.51 and 12.85 and the yields are 6.17% and 2.47%, respectively. MnP1 and MnP2 have approximate molecular masses of 46.3kD and 43.0kD respectively, as determined by SDS-PAGE. The isoenzymes differed in optimum temperature (60degreesC and 65degreesC) and optimum pH(5.8 and 6.2) for oxidation of DMP (2,6-dimethoxyphenol). MnP1 and MnP2 are stable below 45degreesC and ranging from pH4.0 to pH7.0. DMP is the best substrate, the Km values of MnP1 and MnP2 for DMP are 13.43micromol/L and 12.45micromol/L respectively. Catalysis doesn't occur in the complete absence of Mn. EDTA inhibites the activities of MnP1 and MnP2 at the higher concentration and DTT inhibites the enzyme activities completely. PMID- 16342762 TI - [Expression and characterization of FNRD in cyanobacterium Synechococcus sp. PCC 7002]. AB - The petHL genes under the control of Lac and Kan promoters were transformed into Synechococcus sp. PCC 7002, respectively. Both of the petHL genes are integrated into the cyanobacterium chromosomes, which is inferred from the results of Southern blot analysis. Western blot analysis results show that both petHL genes are expressed in the transformed cells, and Kan promoter is more effective than Lac promoter. The FNRD in vivo shows the same stability as that of FNR holoenzyme. Some FNRD molecules are probably acylated as judged by the result of Triton X-114 phase partition test. FNRD in vivo might act as a component in photosynthetic electron transport chain, which increases the photosynthetic oxygen evolution rate. PMID- 16342763 TI - [Cloning, expression and characterization of antibacterial peptide CM4 in Pichia pastoris]. AB - To explore a new approach of expression of ABP-CM4 in methylotrophic yeast P. pastoris. The gene of ABP-CM4 was cloned into the vector pPIC9. The Sac I linearized plasmid pPIC9-CM4 was transformed into P. pastoris GS115 by electroporation. By means of MM and MD plates and PCR, the recombinant P. pastoris strains (his- mut+) were obtained. Antibacterial assay, Tricine-SDS-PAGE and Polyarylamide Gel Electrophoresis demonstrated that ABP-CM4 was extracellularly expressed in P. pastoris with the induction of methanol. The recombinant ABP-CM4 has the antifungal activity on Aspergillus niger and Trichoderma viride. PMID- 16342764 TI - [Study on population diversity and antimicrobial activity of actinomycete from acidic soil in Xizang area]. AB - 156 actinomycetes were isolated from 10 soil samples collected in Tibet by the method of DDC(dispersion and differential centrifugation). The isolates were assigned to 32 different groups on the basis of their morphological characters and colors of substrate mycelium, aerial hyphae and pigment. The result of cell wall DAP on 65 representative strains selected from these groups was that 9 strains were different from Streptomyces. The analysis of ARDRA and 16S rDNA sequeces showed that the isolates belonged to the genus Streptomyces and five different rare actinomycete genera respectively. Antimicrobial activity of the representative strains was also tested, with the result that 38.5% isolates were positive. PMID- 16342765 TI - [Isolation and characterization of a HCH degradation Sphingomanas sp. stain BHC A]. AB - An aerobic bacterium was isolated successfully from a long-term contaminated upland field, which was named BHC-A. The bacterium can utilize Hexachlorocycloexane as a sole carbon source and decompose this substance rapidly and completely. According to its physiological & biochemical characters and the homology analysis of its 16S rDNA sequence, this strain was identified as Sphingomonas sp. Stain BHC-A canmineralize not only alpha, gamma and delta-HCH rapidly, but also 5 mg/L beta-HCH in 12h completely. However, Sphingomonas paucimobilis UT26, a gamma-HCH-decomposing bacterium, can only degrade 12.6% of 5 mg/L alpha-HCH, but not beta-HCH and delta-HCH, even for more than 24h in the medium. The seedling of cucumber was selected as the model plant subject to the inhibition of HCH to the growth of cucurbitaceous plants. The results shows that the degradation of HCH isomers by BHC-A in soil is even steady and excellent, that BHC-A can eliminate the contamination of the soil and the harm to the plants. PMID- 16342766 TI - [Comparison of the desulfurization activity among several bacteria and analysis of the conservation of their desulfurization genes]. AB - Several bacteria, Bacillus brevis R-6, Pseudomonas delafleldii R-8, Nocardia globerula R-9, Bacillus sphaericus R-16, Rhodococcus erythropolis LSSE8-1 and Gordonia nitida LSSEJ-1, which can convert dibenzothiophene into 2 hydroxybiphenyl and sulfate, were investigated. Desulfurization products were quantitively determined by HPLC. Result revealed that each of these bacteria desulfurize DBT at a different rate. In order to obtain more information, the fragments encoding desulfurizing enzymes were studied. Desulfurization genes of R 6 and R-8 were separately amplified via PCR with specific primers based on the related sequences of Rhodococcus sp. IGTS8. Both sequences areminimally 99% related to IGTS8 sequence. Afterwards, dsz operon of LSSEJ-1 and R-9 were amplified and cloned. Sequences are also highly conservative. Data shows that identity of dszA between R-9 and IGTS8 is 99.6%, and identity of dszA between LSSEJ-1 and IGTS8 is 99.9%; dszB sequence of R-9 and LSSEJ-1 is 99.6% similarity to their counterpart sequence from IGTS8;Identity of dszC between R-9 and IGTS8 is 99.9%, and identity of dszC between LSSEJ-1 and IGTS8 is 99.1% . It may be deduced that the origins of desulfurization genes from mesophilic bacteria are the same. PMID- 16342767 TI - [Analysis of Bifidobacteria spp. composition in human gut via temperature gradient gel electrophoresis (TGGE)]. AB - Temperature Gradient Gel Electrophoresis (TGGE) method combined with 16S rDNA clone library profiling was used to analyze Bifidobacteria spp. composition in human gut in this study. Bifidobacteria group-specific TGGE patterns of 10 healthy human individuals showed that the Bifidobacteria population in humans was host-specific. The genomic diversity and species composition of Bifidobacteria in different individual was dissimilar. Through sequencing and TGGE comigration analysis of the Bifidobacteria group-specific amplicons for one healthy boy, it was revealed that the TGGE bands of this individual represented species Bifidobacterium bifidum, B. infantis, B. longum, B. adolescentis, B. pseudocatenulatum and one potentially new species, respectively. B. pseudocatenulatum was the most common species of tested samples. While as control, only two species- B. pseudocatenulatum and B. longum were isolated using traditional culture method. Bifidobacteria group-specific PCR based TGGE method combined with 16S rDNA clone library profiling is a sensitive and effective approach to resolve the population structure of Bifidobacteria in microflora of human intestinal tract. PMID- 16342768 TI - [Stability of recombinant plasmids in attenuated Salmonella typhimurium and their effects on bacterial invasiveness and multiplication]. AB - Effects of plasmid type and insertion sequence on in vitro and in vivo multiplication and invasiveness of attenuated Salmonella typhimurium were examined following transformation of the bacteria with eukaryotic expression plasmids pcDNA3 and pCI with or without heterologous gene (Newcastle disease virus F gene). Exogenous plasmids had negative impacts on the replication or invasiveness of the attenuated S. typhimurium in LB broth and/or HeLa cell monolayers as well as on its survival in live chicks. The plasmid pCI had more significant effects than pcDNA3. Introduction of heterologous gene into the plasmids not only exhibited additional negative influence on the host strain but also on their own stability therein. All these results suggest that full consideration should be given to the types of plasmids and their stability within the host strain as well as to their effects on replication and invasiveness of attenuated bacteria as the DNA vaccine delivery vector for improved immune protection. PMID- 16342769 TI - [Construction and characterization of a live attenuated Shigella flexneri 2a vaccine strain, sf301 Delta virG and dsbA33G]. AB - Construction and characterization of a live attenuated Shigella flexneria 2a sf301 vaccine strain to prevent the endemic of shigellosis. Using Chinese majority epidemic Shigella flexneri 2a serotype sf301 as the target, p Delta virG, a deletion derivation of the virG gene in the SacB suicide vector pCVD442 and pDsbA33G, an mutant of a disulfide bond catalyst DsbA, replaced its 33 amiano acid Cystine by Glycerin in pCVD442, were used to generate a attenuated mutant strain sf301: Delta virG: DsbA33 G. Its virulence was evaluated by Sereny test, the invasive ability was detected by HeLa cell invasive assay, immunogenicity was detected by immunized Guinea pigs through inoculated guinea pigs' conjunctive sac. Sereny test was negative and HeLa invasive assay showed sf301: Delta virG: DsbA33 G retained partial invasive ability. In contrast to control group, sf301: Delta virG: DsbA33 G could induced significantly high antibody levels of IgA and IgG against sf301 LPS in animal's mucosal lavage fluids and sera in both primary immunization protocol and boosting protocol. The numbers of ASCs in local draining lymph nodes and spleens were significantly higher than control group. The immune response to sf301: Delta virG: DsbA33 G could provide completely protection from the challenge of wild type sf301. sf301: Delta virG: DsbA33 G is a safe and effective oral candidate vaccine to prevent the infection of Shigella strains. PMID- 16342770 TI - [Determination of fibronectin-binding region of FBPS of Streptococcus suis type 2]. AB - Fusion expression plasmid pFBPS, pFBPS (7 - 82), pFBPS (7 - 165) and pFBPS3 (87 - 320) of Streptococcus suis type strain HA9801 were generated by cloning different fragments of fbps amplified from pMD-T-FBPS by PCR into plasmid pET-32a(+). It has been confirmed that the recombinant proteins, FBPS, FBPS(87 -320), and FBPS(7 - 167), which are expressed by recombinant plasmid pFBPS, pFBPS(87 - 320) and pFBPS(7 - 165), respectively, bound human fibronectin by ligand affinity Western blot assay. The results indicate the primary fibronectin-binding domain of FBPS lies within 87 - 165 amino acid residues region. PMID- 16342771 TI - [Study on infection of different strains of Vibro cholerae O1 by El Tor CTXPhi]. AB - To study the horizontal transfer efficiencies of filamentous bacteriophage CTXPhi in different V. cholera O1 strains and the phage immunities of these strains. The infectious El Tor CTXPhi particles genetic marked by chloramphenicol resistance gene were used to infect four different V. cholerae O1 strains in vivo and in vitro. Selected the infected clones based on its character of chloramphenicol resistance and identified and judged the exist form of CTXPhi genome through Southern bolt and other hybridization methods. Calculated the infection rates of different strains and compared each other. Then we analyzed the mechanism of infection and phage immunity. The infection rate of classic strain 1119 with the genetic marked CTX(ET)Phi in vivo is much higher than that in vitro. In vitro experiment, the rate of 1119 is higher than other three El Tor strains. And in El Tor strains, the infection frequency of IEM101 that had no rstR gene is 100 to 1000 times higher than other two strains containing rstR. Classical biotype strain is more susceptible to CTX(ET)Phi particles than El Tor strains. Expression of TCP and the phage immunity mediated by rstR gene affect the horizontal transfection of CTXPhi in V. cholerae strains. PMID- 16342772 TI - [Cloning, expression and preparation of polyclonal antibody for IBDV non structure protein gene]. AB - Infectious bursal disease virus (IBDV) VP5 gene was amplified and cloned into an N-terminal GST-Tag fusion expression vector, pGEX-4T-2, which was controlled by T7 promoter. The sequencing result showed that the VP5 gene was composed of 438 base pairs, and coded 145 amino acids. High VP5 product was expressed in E. coli BL21 induced by IPTG, and the GST-VP5 fusion protein existed in inclusion. High titer anti-VP5 serum was also prepared in New Zealand rabbit immunized with purified fusion protein inclusion. These results gave a basis for further research for VP5 function in IBDV replication and pathogenicity, which also paved the way for developing VP5 gene deleted IBDV live vaccine. PMID- 16342773 TI - [Cloning and expression of outer membrane protein gene Gpd from Treponema pallidum and preliminary studies on its immunogenicity in rabbits]. AB - To construct the recombinant plasmid of Eukaryotic expression containing Gpd gene from Treponema Pallidum and study its immunogenicity in New Zealand White rabbits. Gpd gene was amplified from the genomic DNA of T. pallidum and cloned into appropriate site of pcDNA3. 1 ( + ) vector. After verified that the Gpd antigen gene could be expressed in HeLa cells by Western blot and immunocytochemistry, recombinant plasmids pcDNA3.1 ( + )-Gpd, control plasmid pcDNA3. 1 ( + ) or PBS buffer were administered in three groups of New Zeal and White rabbits. Booster immunizations were employed at 2-week interval for three times. ELISA was used for the quantitative detection of the specific antibody in the sera of rabbits. The proliferation response of spleen cells was detected by MTT assay. The results of the Western blot and immunocytochemistry showed that Gpd gene constructed in pcDNA3.1 ( + ) vector could express a fusion protein with a calculated molecular mass of 41kD in HeLa cells and react with positive blood serum from syphilis patients. The significant specific antibody IgG titers were observed and the highest titer was 1:1024 in rabbits after three times with pcDNA3.1 ( + )-Gpd. The proliferation response of spleen cells were significantly higher than that of rabbits injected with pcDNA3.1 ( + ) (p < 0.05). All above results establish a solid basis for future studying the biological activities of Gpd and benefit the development of the Syphilis DNA vaccine. PMID- 16342774 TI - [Isolation and identification of a photosynthetic bacteria producing coenzyme Q10]. AB - 13 isolates producing Coenzyme Q10 (CoQ10) of purple non sulfur photosynthetic bacteria were enriched out of pond sludge, one isolate named 2c was selected based on its high CoQ10 content and identified systematically. The gram-negative and short-rod shaped strain 2c is 0.6microm - 0.9microm x 1.2microm - 2.0 microm, has one long flagellum on one end of the cell and contains lamellar intracytoplasmic membrane(ICM) system parallel to cytoplasmic membrane. Cultures produce red pigments in the light. Live cells under phototrophic conditions contain bacteriochlorophyll a and carotenoids. 2c grows anaerobically in the light and aerobically in the dark. Optimal growth occurs at 30degreesC - 35degreesC and at pH7.0 -pH8.0. Various organic compounds are used as photosynthetic electron donors and carbon sources. Peptone and (NH4 )2SO4 are its better nitride source,yeast extracts stimulates its growth. A phylogenetic analysis based on 16S rDNA gene sequences reveales that strain 2c gathers a cluster with 3 strains of Rhodopseudomonas palustris whose accession number in GenBank are AY751758, DQ001155, DQ001158, respectively. 2c subcultures 15 generations stably at least. The results presented here demonstrated strain 2c is Rhodopseudomonas palustris. PMID- 16342775 TI - [Screening and identification of steroidal saponins-producing endophytes from Paris polyphylla var. chinensis Franch]. AB - Two endophytic strains SS01 and SS02 with the potential for producing steroidal saponins were isolated from the underground stems of Paris polyphylla var. chinensis Franch. The TLC comparison indicated that there are 3 sports with similar R(f) between the metabolites of SS01 and the saponins of Paris polyphylla var. chinensis Franch. And there are 2 sports with similar R(f) between the metabolites of SS02 and the saponins of Paris polyphylla var. chinensis The and that biochemical SS01 Franch. characteristics of morphology, physiological belonged showed to Enterobacteriaceae and SS02 belonged to Bacillus sp.. The 16S rDNA of SS01 and SS02 were PCR and sequenced. The accessions of GenBank are AY842143 and AY842144, respectively. The two 16S rDNA phylogenetic trees were constructed by comparing with the published 16S rDNA sequences of the relative bacteria species. In the first phylogenetic tree SS01 and Cedecea davisae DSM 4568 was the closest relative with 98.9% sequence similarity, and in the second phylogenetic tree SS02 and Paenibacillus daejeonensis was the closest relative with 97.7% sequence similarity. According to the phylogenetic analysis they were identified as Cedecea davisae SS01 and Paenibacillus daejeonensis SS02, respectively. PMID- 16342776 TI - [Generation of newcastle disease virus strain ZJI isolated from an outbreak in the goose using reverse genetics technique]. AB - The full-length cDNA clone, NDV3GM122, and the three helperplasmids pCI-NP, pCI-P and pCI-L of Newcastle disease virus strain ZJI isolated from an outbreak in the goose were cotransfected into BSR-T7/5 cell expressing T7 RNA polymerase. Meanwhile, the full-length cDNA clone NDV3GM122 and the three helperplasmids, pCIneoNP, pCIneoP and pCIneoL which were derived from NDV strain La Sota, were also cotransfected into the cell, respectively. Indiect immunofluorescence assay (IFA) was performed 48 to 96 hours post-transfection using NDV HN-specific monoclonal anbtibody (McAb) 6B1 and bright stainings were found in the transfectants, indicating that the full-length clone was functional and the HN protein was expressed. The transfected cell and the supernatant were mixed well and thereafter the mixture was inoculated into specific pathogen free (SPF) chicken eggs. The allanotoic fluid of the injected eggs gave a positive hemagglutinin( HA) titer ranging from 16 to 32 in the secondary passage and increased to 128 in the third passage, which was same to the level of parent wild type virus. The allantoic fluid containing the recovered NDV was analyzed in hemagglutination inhibition( HI) test by using McAb 6B1 and the specific inhibition was found. The typical morphology of the produced NDV was detected in the electronic microscope. The results mentioned above demonstrated that infectious NDV of strain ZJI was successfully generated, which laid good foundation for the further related research. PMID- 16342777 TI - [Agrobacterium tumefaciens-mediated transformation of the white-rot basidiomycete, phanerochaete chrysosporium]. AB - Agrobacterium tumefaciens has been used to transfer its T-DNA to filamentous fungus Phanerochaete chrysosporium. A DNA fragment containing recombinant hygromycin B phosphotransferase gene (rhyg(r)) which is controlled by a promoter from P. chrysosporium and the lignin peroxidase gene terminator was cut out from pCH6, and inserted into a plant binary plasmid pCAMB1300, resulted in the recombinant plasmid pCH6-1300. The plasmid pCH6-1300 was then transformed into Agrobacterium tumefaciens strain A208, which was used to infect conidia or protoplasts of P. chrysosporium. Sixteen hygromycin B-resistant P. chrysosporium colonies were obtained. The transformation frequency is about 8 - 9 transformants per 106 conidia or protoplasts, which had no significant difference with the values obtained from conidia or protoplasts. One of the transformants named AT18, whose colony morphology was different from the wild-type strain. The results from both PCR and Southern blot analysis of transformants demonstrated that the rHyg(r) gene was integrated into the host genome with single copy. This approach may not require any instrument and digestion of fungal cell walls. A . tumefaciens-facilitated transformation should be very useful for further studies of the molecular genetics of P. chrysosporium. PMID- 16342778 TI - [Expression, purification and antigen activity analysis of the N-terminal domain of lipoprotein LppQ of Mycoplasma mycoide subsp. mycoides SC]. AB - The gene sequence coding the N-terminal domain of LppQ was amplified from Mycoplasma mycoides subsp. mycoides SC (MmmSC) HVRI X strain by PCR using special primers and was cloned into the EcoR I /Sal I sites of pET32a vector to construct the expression recombinant plasmids. The recombinant plasmids were indentified by restriction digestion, PCR and sequence analysis. The gene was overexpressed in Escherichia coli BL21 (DE3) host cell and the soluble protein was purified with Ni-NTA His. Bind purification kits. The amount of recombinant protein reached 53.7% of the total mass of bacterial protein. The purity of recombinant protein reached to over 95 %. The antigen activity of the purified protein was examined with Western blot analysis. The purified protein reacted strongly with the standard positive serum and didn't react with the negative sera of contagious bovine pleuropneumonia(CBPP). PMID- 16342779 TI - [Expression of B-glucosidase gene of Sacchromycopsis fibuligera in Pichia pastoris]. AB - A beta-Glucosidase gene (BGL 1) was amplified with PCR from the total DNA of Sacchromycopsis fibuligera, and was linked with pGEM-T vector. After cut down by restriction enzyme from pGEM-T vector, BGL 1 was inserted into the expression vector pPIC9K of Pichia pastoris in reading frame with alpha-factor secreting signal peptide sequence to construct the recombinant plasmid pSHL9K. The recombinant plasmid pSHL9K was transformed into Pichia pastoris GS115 with electroporation. The recombinant Pichia pastoris strains which could efficiently secret recombinant beta-Glucosidase were selected. The optimum temperature of the recombinant beta-Glucosidase was 50degreesC, and the optimum pH was 5.4. The activity of beta-Glucosidase could reach to 47U/mL in the culture medium. PMID- 16342780 TI - [Accumulation of carotenoid in Colletotrichum gloeosporioides induced by blue light]. AB - Blue light induced carotenoid accumulation in filamentous fungus- Colletotrichum gloeosporioides. The pigment particles were observed in the fungal mycelium by microscope after blue light irradiation. The pigmentation was not found in the fungal mycelium either cultured under dark condition or cultured in red light. The accumulation of the carotenoid was regulated by blue light intensity. When C. gloeosporioides was incubated in blue light of 6.5micromol x m(-2) x s(-1) under 28degreesC, carotenoid content increased with the irradiation time and reached to a peak of 71.8microg/g FW at 5d. The treatment of dark pre-culture of C. gloeosporioides also had effect on the production of the pigment induced by blue light. PMID- 16342781 TI - [Microbial growth kinetics model of specific spoilage organisms and shelf life prediction for tilapia at fluctuating temperatures]. AB - It was studied on the growth kinetics model of specific spoilage organisms Pseudomonas spp. for cultured tilapia during aerobic storage at fluctuating temperatures from 0degreesC to 15degreesC and the applicability of the model in predicting the remaining shelf life. The mathematical model based on the effect of temperature on Pseudomonas spp. growth kinetics was developed by a Belehradek type equation. Bias and accuracy factors range from 0.906 to 0.942 and from 1.13 to 1.19, respectively, by comparing predicted value using Pseudomonas spp. growth kinetics model with observed value of Pseudomonas spp. growth for cultured tilapia stored at fluctuating temperatures under two kinds of fluctuating temperature designed . Relative errors by comparing remaining shelf life predicted based on growth model of Pseudomonas spp. with remaining shelf life experimentally determined by analyzing organoleptic, VBN and the number of Pseudomonas spp. on tilapia are 5.9 % and - 9.1%, respectively. It shows that the growth kinetics model of Pseudomonas spp. is valuable for rapid and realistic remaining shelf life prediction of cultured tilapia stored aerobically at fluctuating temperatures from 0- 15 degrees C. PMID- 16342782 TI - [Preliminary study on bacterial culture through near-natural pure culture technique]. AB - Based on the ecological features of mutual benefits of different microbes in natural environment, near-natural pure culture technique which was an improvement of conventional pure culture method had been put forward. The key point of this new method was to make hole petri dishes covered with millipore membranes. When these covered dishes being used to culture microorganisms, it could provide incubated microbes active materials from native environment, maintain free communicating with outside microbes, and improve their recovery and culturability of some bacteria. To some degree, this technique enhanced their strengths and overcome their weaknesses of mixed culture method and conventional pure culture method so as to increase microbial culturability and even possibly obtain some nonculturable microorganisms. PMID- 16342783 TI - [Load of calcium probe Fura -2/AM in Escherichia coli cells]. AB - The fluorescence characteristics of Fura-2/AM, a kind of calcium probe normally used in analysis of animal cells, were studied in detail in this work. The load of the probe into Escherichia coli cells was elucidated through fluorescence spectra. The combination of Fura-2 with calcium ions inside Escherichiacoli cells was observed, and the transmembrane behaviors of extracellular calcium to Escherichia coli cells were also investigated. The work was aimed at further researches on the calcium-induced horizontal gene transfer in Escherichia coli cells. PMID- 16342784 TI - [Molecular mechanism of the integration and lysis of mycobacteriophage]. AB - Tuberculosis remains one of the major threats to public health. China is one of the heavy TB burden countries. Novel drugs and vaccines are urgently needed to combat the increasingly multidrug resistant TB. Mycobacteriophage is one of the hot topic in TB novel drugs discovery and drug susceptibility test. Phages can multiply via two alternative mechanisms: the lytic cycle or the lysogenic cycle. The lytic cycle ends with the lysis and death of the host cell, whereas the host cell remains alive in the lysogenic cycle. Lysogenic mycobacteriophages were intensively studied to elucidate the integration and lysis mechanisms of mycobacteriophage. The integration of mycobacteriophage requires for attP of bacteriopahge genome, attB of Mycobacterium genome, integrase and integration host factor. Some lysogenic phage, eg. mycobacteriophage Ms6, employ lytic cycle, form new phage, lysis host by the cooperation of lysin and holin, and release phages. There is no reports as to the mycobacteriophage unique to China clinical or environmental isolates. Studies on the integration and lysis molecular mechanism of mycobacteriophage might facilitate future new anti-TB drugs development. PMID- 16342785 TI - [Newly progress in applications of Saccharomyces cerevisiae cell-surface engineering]. AB - Saccharomyces cerevisiae cell-surface engineering is a newly genetic expressing system for fusion heterologous gene. With its ability of post-translational modification, and the convenience to identify and screen the expressed products, it is undergoing a marvelous progress in its newly applications, which include using as whole-cell biocatalyst, antigen/antibody library, bioadsorbent, biosensor, combinatory protein library, immunoassays and affinity purification. It performs a more and more important role in exploring the function and application of protein molecules. PMID- 16342786 TI - [Wolbachia endosymbionts and their effects on the fitness of the arthropod hosts]. AB - Wolbachia are common and maternally inherited bacteria found in reproductive tissue of a wide range of arthropod species. A tremendous amount of progress on their manipulating reproduction of their host has been made over the past 30 years. Recent surveys have found that they could effect the fitness of their hosts. The recent advances on Wolbachia distribution, locality and their effects on the fitness of hosts are reviewed, and the significance and potential implications of the fields are discussed. PMID- 16342787 TI - Ultratraces analysis of organochlorine pesticides in drinking water by solid phase extraction coupled with large volume injection/gas chromatography/mass spectrometry. AB - This study describes an SPE coupled with large volume injection (LVI) analytical method for the analysis of organochlorine pesticides, BHC (alpha, beta, delta), aldrin, endosulfan (alpha, beta), endrin, dieldrin, and DDT, from aqueous samples. Determination was carried out by GC with MS. The LODs of organochlorine pesticides were determined at 10 ng/L concentration levels, and the results show that SPE-LVI-GC/MS has the potential to accurately determine organochlorine pesticides in water, as it avoids analyte classes in the various steps of a typical extraction procedure. PMID- 16342788 TI - Development and validation of an HPLC confirmatory method for the determination of tetracycline antibiotics residues in bovine muscle according to the European Union regulation 2002/657/EC. AB - A high-performance liquid chromatographic method with diode-array detection, at 351 nm, was developed and validated for the determination of five tetracyclines (TCs): minocycline, tetracycline, oxytetracycline, chlortetracycline, and doxycycline in bovine muscle. Samples were macerated with a buffer solution, centrifuged, and purified using Abselut Nexus SPE cartridges. The separation of the examined TCs was achieved on an Inertsil ODS-3 5 microm, 250 x 4 mm analytical column, at ambient temperature. A multistep gradient elution was followed using 0.05 M oxalic acid and CH3CN, at a flow rate of 1.65 mL/min. The procedure was validated according to the European Union regulation 2002/657/EC determining selectivity, stability, decision limit, detection capability, accuracy, and precision. The results of the validation process demonstrate that the method can be readily applied to European Union statutory veterinary drug residue surveillance programmes. Mean recoveries of TCs from bovine muscle samples spiked at three concentrations (100, 250, and 400 ng/g) were in the range of 98.7-103.3%. Method's LOQ values achieved were 40 microg/kg for MNC, CTC, and DC and 25 microg/kg for OTC and TC. The decision limits (CCalpha) were in the range of 104.7-109.8 microg/kg, while the detection capability (CCbeta) was in the range of 108.4-116.7 microg/kg for all compounds. PMID- 16342789 TI - Online trace enrichment to determine pyrethroids in river water by HPLC with column switching and photochemical induced fluorescence detection. AB - The potential of online trace enrichment on a highly apolar short column in LC was evaluated for the determination of pyrethroids in river water. Twelve millilitres of water samples, modified with 8 mL ACN (ACN/water 40:60, v/v), were passed through 50 x 4.6 mm ID first separation column packed with 5 microm Hypersil Elite C18. Pesticides were preconcentrated in this column while the matrix background was eluted to waste. Separation of pesticides was performed on a 3.5 microm symmetric C18 column (250 x 4.6 mm ID) with an ACN step gradient as mobile phase and fluorescence detection was used after postcolumn derivatization by using UV light. The use of photochemically induced fluorescence for detection improved sensitivity and selectivity. Quantification limits ranged from 0.05 to 0.1 microg/L and pesticide recoveries at two concentration levels (0.1 and 0.5 microg/L) were between 93.1 and 118.6%, with RSD between 2.5 and 7.5% (n = 3) in river water samples. No matrix effect was detected. PMID- 16342790 TI - HPLC isolation and NMR structure elucidation of the most prominent octabromo isomer in technical octabromo diphenyl ether. AB - The major octabromo isomer of technical octabromo diphenyl ether mixture (technical octaBDE) DE-79 was isolated by RP-HPLC. Three serially coupled columns (each 250 mm long) enabled a good separation of the target compound from other congeners using 100% ACN as eluent. Approximately 100 microg of the target compound was isolated with a purity of >90% and investigated by MS for confirmation of the degree of bromination. 1H-NMR and 2-D 1H-13C correlation NMR spectra unequivocally clarified that the octaBDE in question is 2,2',3,3',4,4',6,6'-octabromodiphenyl ether (BDE 197). Based on annual production rates of technical BDE products (1999/2001), approximately 380 tons of BDE 197 were annually produced which, on the long term, may enter the environment. Compared with other individual BDE congeners, BDE 197 has the seventh highest application rate. Reductive debromination of BDE 197 can lead to four hepta-, 15 hexa-, 23 penta-, and 28 tetra-BDE isomers, respectively. This variety includes all known major BDEs of environmental concern (BDE 47, 85, 99, 100, 153, 154, and 183). The identification of BDE 197 in technical octaBDE DE-79 strongly suggests that research on the environmental fate of BDEs should include this key-BDE congener. PMID- 16342791 TI - High performance liquid chromatographic separation of dipeptide and tripeptide enantiomers using a chiral crown ether stationary phase. AB - A chiral stationary phase (CSP) based on (-)-(18-crown-6)-2,3,11,12 tetracarboxylic acid was evaluated for the direct resolution of the enantiomers of dipeptides and tripeptides. The type and concentration of the acid and the methanol content were optimized with regard to retention time and resolution using Ala-Phe as model peptide. A mobile phase consisting of 10 mM sulfuric acid in 70% aqueous methanol was applied to the separation of a set of 16 structurally diverse dipeptides and tripeptides. Generally, the configuration of the amino acid at the N-terminus determined the enantiomer elution order. With a few exceptions the LL- and LD-enantiomers interacted stronger with the CSP compared to the corresponding DD- or DL-enantiomers. The experimental conditions also allowed the simultaneous separation of all four stereoisomers of Ala-Phe. Addition of ammonium sulfate generally reduced retention times and enantiomer resolution. Addition of triethylamine as modifier led to an overall increase of the retention times while the resolution did not show a general trend, increasing in the case of Ala-Ala but decreasing in the case of Ala-Phe. PMID- 16342792 TI - Chiral separation of 1,1'-bi-2-naphthol and its analogue on molecular imprinting monolithic columns by HPLC. AB - Two molecular imprinting polymer (MIP) monolithic columns with (S)-(-)-1,1'-bi-2 naphthol and (R)-(+)-5,5',6,6',7,7',8,8'-octahydro-1,1'-bi-2-naphthol as the templating molecules, respectively, have been prepared by in situ polymerization using 4-vinylpyridine and ethylene dimethacrylate as functional monomer and cross linker, respectively. The columns with good flow-through properties were obtained by changing the molar ratio of the functional monomer and the template molecule. The effects of mobile-phase composition on separation of enantiomers were systematically investigated. The results indicate that hydrophobic interaction in aqueous solution and hydrogen-bonding interaction in ACN between the enantiomers and polymers could play important roles in the retention and resolution. The effects of chromatographic conditions, such as flow rate, column temperature, sample loading, on the enantioseparation were also studied. Further, these two MIP columns show a cross-reactivity. PMID- 16342793 TI - Simultaneous densitometric determination of artemisinin, artemisinic acid and arteannuin-B in Artemisia annua using reversed-phase thin layer chromatography. AB - A rapid and simple RP-TLC method for simultaneous quantification of pharmacologically important sesquiterpene artemisinin (AM) together with its precursors arteannuin-B (AB) and artemisinic acid (AA) in the inflorescence part of Artemisia annua plant has been developed. The RP-TLC of sesquiterpenes was performed on RP-18 F254 S thin-layer chromatographic plates by developing in mobile phase, containing 0.2% TFA in water/ACN (35:65, v/v). The densitometric determination of AM, AB and AA was carried out after derivatization with anisaldehyde reagent at 426 nm in absorption-reflectance mode. PMID- 16342794 TI - Extraction and analysis of different Cannabis samples by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry. AB - A headspace solid-phase microextraction combined with GC-MS method was developed for the extraction and analysis of cannabinoids from Cannabis samples. Different commercially available fibres were evaluated; polydimethylsiloxane 100 microm was selected as the most efficient one. In order to enhance sensitivity and reduce analysis time, the sampling temperature was studied and it showed that extraction should be performed at a high temperature (150 degrees C). In relation with the high lipophilicity of cannabinoids, a relatively long desorption time (3 min) was necessary to ensure a total transfer from the fibre into the injection port of the gas chromatograph. The method was finally applied to the extraction of Swiss marijuana samples from different regions. Data treatment by principal component analysis and hierarchical cluster analysis allowed a discrimination of the different batches. PMID- 16342795 TI - Headspace single-drop microextraction with in-drop derivatization for aldehyde analysis. AB - A new technique, headspace single-drop microextraction (HS-SDME) with in-drop derivatization, was developed. Its feasibility was demonstrated by analysis of the model compounds, aldehydes in water. A hanging microliter drop of solvent containing the derivatization agent of O-2,3,4,5,6 (pentaflurobenzyl)hydroxylamine hydrochloride (PFBHA) was shown to be an excellent extraction, concentration, and derivatization medium for headspace analysis of aldehydes by GC-MS. Using the microdrop solvent with PFBHA, acetaldehyde, propanal, butanal, hexanal, and heptanal in water were headspace extracted and simultaneously derivatized. The formed oximes in the microdrop were analyzed by GC-MS. HS-SDME and in-drop derivatization parameters (extraction solvent, extraction temperature, extraction time, stirring rate microdrop volume, and the headspace volume) and the method validations (linearity, precision, detection limit, and recovery) were studied. Compared to liquid-liquid extraction and solid-phase microextraction, HS-SDME with in-drop derivatization is a simple, rapid, convenient, and inexpensive sample technique. PMID- 16342796 TI - High extraction efficiency fiber coated with calix[4] open-chain crown ether for solid-phase microextraction of polar aromatic and aliphatic compounds. AB - The calix[4] open-chain crown ether, 5,11,17,23-tetra-tert-butyl-25,27-di(2 allyloxyethoxyl)-26,28-dihydroxycalix[4]arene was synthesized and used for preparation of solid-phase microextraction (SPME) fibers of enhanced extraction efficiency. The new SPME coating made from calix[4] open-chain crown ether and hydroxyl-terminated silicone oil was developed with the aid of vinyltriethoxylsilane as bridge using sol-gel method and cross-linking technology. The efficiency of the novel fiber in the extraction of polar aromatic and aliphatic compounds, such as phenols, alcohols, and volatile fatty acids, was also investigated. Due to the introduction of the polar open-chain crown ether in calix[4]arene molecules, the calix[4] open-chain crown ether fiber showed much better selectivity and sensitivity to these polar compounds in comparison with calix[4]arene fiber. It also had superior extraction efficiency when compared to commercial poly(dimethylsiloxane)-divinylbenzene and polyacrylate fibers. Parts per billion to parts per trillion level detection limits were achieved for most of the analytes through SPME in conjunction with GC and flame ionization detector. The linear ranges were two to four orders of magnitude, and the RSD values were below 7% for all analytes. The novel fiber was applied to determine volatile alcohols and fatty acids in wine samples. The volatile-free wine prepared in this work was used to assure similar chemical environment for analytes in both calibration solutions and in real wine samples, thus compensating for possible matrix interferences. The established internal standard method using 4-methyl-2-pentanol as internal standard showed satisfactory accuracy and precision. PMID- 16342797 TI - Near-critical carbon dioxide extraction and liquid chromatography determination of UV filters in solid cosmetic samples: a green analytical procedure. AB - Near-critical carbon dioxide extraction of four UV filters used as sunscreens in lipsticks and makeup formulations is reported. Extraction parameters were optimized. Efficient recoveries were obtained after 15 min of dynamic extraction with a 80:20 CO2/ethanol mixture at 300 atm and 54 degrees C, using a 1.8 mL/min flow rate. Extracts were collected in ethanol, and appropriately diluted with ethanol and 1% acetic acid to obtain a 70:30 v/v ethanol/1% acetic acid solution. The four UV filters were determined by LC with gradient elution using ethanol/1% acetic acid as mobile phase. The accuracy of the analytical procedure was estimated by comparing the results with those obtained by methods based on classical extraction. The proposed method only requires the use of CO2, ethanol and acetic acid avoiding the use of more toxic organic solvents, thus it could be considered as both operator and environment friendly. PMID- 16342798 TI - Absolute enantioselective separation: optical activity ex machina. AB - The paper describes methodology of using three independent macroscopic factors affecting molecular orientation to accomplish separation of a racemic mixture without the presence of any other chiral compounds, i. e., absolute enantioselective separation (AES) which is an extension of a concept of applying these factors to absolute asymmetric synthesis. The three factors may be applied simultaneously or, if their effects can be retained, consecutively. The resulting three mutually orthogonal or near orthogonal directors constitute a true chiral influence and their scalar triple product is the measure of the chirality of the system. AES can be executed in a chromatography-like microfluidic process in the presence of an electric field. It may be carried out on a chemically modified flat surface, a monolithic polymer column made of a mesoporous material, each having imparted directional properties. Separation parameters were estimated for these media and possible implications for the natural homochirality are discussed. PMID- 16342799 TI - Enantiomeric composition studies in Lavandula species using supercritical fluids. AB - Characteristic aroma compounds in plants and essential oils of Lavandula from different varieties were examined. The study of the qualitative and quantitative composition of the major volatile components was faced by developing a method based on the use of supercritical fluid extraction-GC-MS (SFE-GC-MS). The optimization of a variety of parameters affecting SFE extraction enabled RSDs from three replicates lower than 2% to be achieved. Equally, recoveries of up to 59% were obtained by applying the proposed method. The use of multidimensional GC was necessary to enantiomerically resolve the target compounds. The obtained results showed enantiomeric purities >90% for all studied compounds in all varieties considered, proving the natural invariability of the enantiomeric composition of the compounds of interest. This information can be useful in authenticity studies as well as in selecting natural sources of enantiomerically pure compounds. PMID- 16342800 TI - Application of multiwalled carbon nanotubes as solid phase extraction sorbent for preconcentration of trace copper in water samples. AB - The adsorption behavior of multiwalled carbon nanotubes (MWNTs) toward copper has been investigated systemically, and a new method has been developed for the determination of trace copper in water samples based on preconcentration with a microcolumn packed with MWNTs prior to its determination by flame atomic absorption spectrometry. The optimum experimental parameters for preconcentration of copper, such as pH of the sample, sample flow rate and volume, elution solution and interfering ions, have been investigated. Copper can be quantitatively retained by MWNTs in the pH range 5-8, and then eluted completely with 0.5 M HNO3. The detection limit of this method for Cu was 0.42 ng/mL, and the RSD was 3.5% at the 10 ng/mL Cu level. The method was validated using a certified reference material, and has been successfully applied for the determination of trace copper in water samples. PMID- 16342801 TI - Optimization of a new resin, Amberlyst 36, as a solid-phase extractor and determination of copper(II) in drinking water and tea samples by flame atomic absorption spectrometry. AB - A new simple and reliable method has been developed to separate and preconcentrate trace copper ion in drinking water and tea samples for subsequent measurement by flame atomic absorption spectrometry (FAAS). The copper ions are adsorbed quantitatively during passage of aqueous solutions through Amberlyst 36 cation exchange resin. After the separation and preconcentration stage, the analyte was eluted with a potassium cyanide solution and determined by FAAS. Different factors including pH of sample solution, sample volume, amount of resin, flow rate of aqueous solution, volume and concentration of eluent, and matrix effects for preconcentration were examined. The analytical figures of merit for the determination of copper are as follows: analytical detection limit (3 sigma), 0.26 microg/L; precision (RSD), 3.1% for 100 microg/L; enrichment factor, 200 (using 1000 mL of sample solution and 5 mL of eluent); time of analysis, 3.5 h (for obtaining enrichment factor of 200); capacity of resin, 125 mg/g. The method was applied for copper determination by FAAS in tap water, commercial natural spring water, commercial treated drinking water, and commercial tea bag sample. The accuracy of the method is confirmed by analyzing tea leaves (GBW 07605). The results demonstrated good agreement with the certified values. PMID- 16342802 TI - Novel RPLC stationary phases for lipophilicity measurement: solvatochromic analysis of retention mechanisms for neutral and basic compounds. AB - An RPLC was developed to rapidly determine lipophilicity of neutral and basic compounds using three base deactivated RPLC stationary phases particularly designed for the analysis of basic compounds, namely, Supelcosil ABZ(+)Plus, Discovery RP Amide C16, and Zorbax Extend C18. The work consisted of three sets of experiments. In the first log kw values of neutral compounds were extrapolated using hydroorganic mobile phases at different compositions. Good correlation between log kw and log Poct indicated that the method was appropriate for these supports, without adding a silanol masking agent. In the second set of experiments, isocratic log k values of neutral and basic compounds were measured with three different mobile phases. The best estimation of lipophilicity was obtained for neutral and basic compounds when the secondary interactions were strongly reduced (i. e., when basic compounds were under their neutral form). In the third set of experiments, isocratic retention factors of basic compounds (in their neutral form) were measured with a high-pH mobile phase, on a chemically stable support (Zorbax Extend C18). Under these chromatographic conditions, correlation between the isocratic retention factors and log Poct (log D10.5) for basic compounds was similar to that for neutral compounds. PMID- 16342803 TI - A cyanopropyl reversed phase chromatography enables resolution of huperzine A and B on analytical and preparative scales. AB - Huperzine A (HupA) and huperzine B (HupB) are two medically important components of Huperzia serrata. It is difficult to obtain high yields of the separation from the plant using conventional liquid extraction and chromatography. However, this study has found that RP chromatography with cyanopropyl (CN) medium was able to separate these two analogs simultaneously from the plant extract with higher resolution. Comparison between a CN medium and a popular C18 medium demonstrated the superiority of the CN over the C18 in resolution for both analytical and preparative separation of HupA and HupB. A preparative process was developed for simultaneous purification of HupA and HupB from H. serrata. The yields on the basis of the mass of the herbal powder for HupA and HupB were 0.019 and 0.008% respectively, which were about 1.9 and 10 times of those reported in the literature. PMID- 16342804 TI - Determination of some polycyclic aromatic hydrocarbons in filter tar of Turkish cigarettes. AB - A simultaneous determination method for the analysis of benzo[a]pyrene (BaP) and dibenzo[a,h]anthracene (DahA) in the filter tar of Turkish cigarettes has been developed. The method involved (a) the extraction of BaP and DahA with n-hexane from ACN solution in which the cigarette filters were extracted, and then (b) purification of the n-hexane extracts by elution on an XAD-2 column using n hexane/dichloromethane (9:1, v/v) mixture. Separation and quantitative determination of BaP and DahA in the extracts were carried out by HPLC and fluorescence detection on a C18 RP column. The calculated recoveries for BaP and DahA were found in the range of 90-100% for each extraction and clean-up steps. Analysis of various filter tar of Turkish cigarettes showed that an average of 74.28 ng/filter of BaP and 5.24 ng/filter of DahA were present in Turkish cigarettes. PMID- 16342805 TI - Determination of pKa values by capillary zone electrophoresis with a dynamic coating procedure. AB - CZE allows to measure the acidic dissociation constant (pKa) of many drug substances. However, determining the EOF intensity may be time-consuming, especially at a low pH. In order to overcome this drawback, a dynamic coating procedure of the capillary was carried out to increase microEOF, and thus to reduce the analysis time. In addition, this coating procedure enhanced migration time stability. The effective mobilities of 15 compounds were measured at different pH, producing pK'a values dependent on BGE ionic strength. The latter values were corrected with the activity coefficient to obtain a "true" pKa value. The 15 investigated compounds were (i) five acids: namely, salicylic acid, benzoic acid, ketoprofen, phenobarbital, and phenol, (ii) four bases: lidocaine, propafenone, propranolol, and quinine, (iii), five ampholytes: sulfanilamide, sulfabenzamide, sulfadimethoxine, sulfadoxine, and sulfisoxazole, and (iv) one zwitterion: cetirizine. The range of determined pKa values was between 1.2 and 11.2, and close to the pKa values available from the literature. PMID- 16342806 TI - Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking. AB - CE conditions for monitoring the unsaturated disaccharides of hyaluronic acid (di HA) and chondroitin sulfate (di-CS) using an alkaline tetraborate buffer, electrokinetic sample injection, and UV absorption detection at 232 nm are reported. Separations were performed in an uncoated fused-silica capillary having reversed polarity and reversed electroosmosis generated with the addition of CTAB to the buffer. The influence of various separation parameters, including the concentration of CTAB, buffer pH, concentration of tetraborate, and applied voltage, on the resolution of the two disaccharides was investigated. Baseline separation was obtained with 25 mM tetraborate at pH 10.0 and having 0.05 mM CTAB. Chloride and phosphate in the sample are beneficial for the stacking of the disaccharides, with di-HA forming a much sharper peak than di-CS. Using samples prepared in 25 mM Tris-HCl (pH 7.5) and electrokinetic injection at the cathode at -10 kV for 40 s, linear relationships between the corrected peak area and the concentration of the disaccharides have been found in the ranges of 1.0-400.0 and 0.1-1.0 microg/mL (0.2-1.0 microg/mL for di-CS), with correlation coefficients being >0.9933 in all cases. The RSDs of detection times and corrected peak areas were between 1.13-1.24 and 1.57-2.13%, respectively. Applied to human serum samples that were prepared by ethanol precipitation and depolymerization of the two polysaccharides with chondroitinase ABC reveals comigration of endogenous compounds with di-HA and a sample-dependent detection time. The di-HA content in the serum sample can be estimated via subtraction of the blank peak that is obtained without enzymatic hydrolysis. PMID- 16342807 TI - Ionene-dynamically coated capillary for analysis of urinary and recombinant human erythropoietin by capillary electrophoresis and online electrospray ionization mass spectrometry. AB - In this article, a series of ionene polymers were synthesized and used to coat fused-silica capillaries for the separation of recombinant and urinary human erythropoietin (rhEPO and uEPO) standards by CE. The influence of the charge density of coatings on the separation of rhEPO and uEPO glycoforms was investigated. Then, we further studied the method for fast separation and detection of rhEPO and uEPO standards by CE-ESI-MS. The influence of several CE and MS operating parameters, such as the concentration of CE running buffer, applied external pressure, and the composition and flow rate of sheath liquid on CE-ESI-MS was studied. The results demonstrated that when the capillary was permanently coated with 6,6-ionene and the pH value of acetic acid-ammonium acetate running buffer was 4.80 and 5.50, respectively, a significantly reproducible separation was achieved for rhEPO and uEPO glycoforms. In the online CE-ESI-MS experiments, we not only achieved the online MS signal of uEPO, but also obtained baseline separation of three major rhEPO glycoforms successfully and reproducibly on the 6,6-ionene-coated capillaries. Furthermore, the standard mixture of rhEPO and uEPO was separated, and two incompletely resolved peaks that were identified to be rhEPO and uEPO by the unique MS "fingerprint" were obtained. Additionally, the molecular weight of rhEPO and uEPO were verified and compared to the results by MALDI-TOF-MS. It can be concluded that, in contrast to other indirect methods, the online CE-ESI-MS technique with the combination of the advantages of both CE and MS shows great potential for the separation and detection of rhEPO doping directly in competitive sports. PMID- 16342808 TI - Standard additio--a way of improving quantification of banned azo dyes in leather. AB - An analytical procedure based on supercritical-fluid extraction and microwave assisted extraction was applied on six different real leather samples for the determination of banned azo dyes. Determination of the dyes was performed indirectly by measuring their corresponding harmful aromatic amines, formed after reduction. A comparative study between external standard calibration and standard addition using both the dyes as well as the corresponding amines showed that the latter quantification method provided the highest accuracy. PMID- 16342809 TI - Surface derivatization of poly(p-phenylene terephthalamide) fiber designed for novel separation and extraction media. AB - The surface derivatization of poly(p-phenylene terephthalamide) fiber was studied. The obtained surface-derivatized filaments were packed into a fused silica capillary to evaluate its surface characteristics by using GC. As the stationary phase for GC the surface-derivatized fibers showed higher retention for alkanes and alkylbenzenes than that with the untreated Kevlar fiber. The improvements on the retention power and the peak shape were observed on the benzyl-modified fibrous stationary phase. The derivatized fibrous materials were also evaluated as the extraction medium in fiber-in-tube SPE, and the effect of the surface modification on the extraction power was compared to the parent fiber. The results indicated that the modified fiber possessed a higher extraction power than the untreated fiber. Based on the facts, the successful modification of the fiber surface was estimated. PMID- 16342811 TI - Quadrex corporation. PMID- 16342810 TI - Determination of nonsteroidal antiinflammatory drugs in water samples using liquid chromatography coupled with diode-array detector and mass spectrometry. AB - An analytical method for the determination of trace levels of six different nonsteroidal antiinflammatory drugs (NSAIDs) in water samples has been developed and validated. Environmentally relevant pharmaceuticals were chosen according to human consumption in Poland. Final analysis of the target compounds was performed by RP LC-diode-array detection-MS, whereas sample preparation included an SPE step. For this SPE step, a number of packing materials, such as LiChrolut RP-18, calixarene, Strata-X, BAKERBOND Narc-2, BAKERBOND Polar Plus, BAKERBOND styrene divinylbenzene-1, and Discovery DSC-18, were used, and their respective advantages and disadvantages in this study were discussed. The RP-18 phase was found to be the most retentive for all analytes. The detection limits for compounds in surface waters were varied from 0.005 for diflunisal to 0.095 microg/L for ibuprofen. The average recoveries of NSAIDs from the surface water samples ranged from 80 up to 103%. RSD value is relatively low (from 4% for fenoprofen up to 8% for ibuprofen). The performance of the method was tested with several environmental water samples. PMID- 16342812 TI - News you can use. PMID- 16342813 TI - Financing your dream. PMID- 16342814 TI - Interstitial cystitis. Current diagnostic and management strategies. PMID- 16342815 TI - Heart failure management. Prevention of pulmonary edema is essential. PMID- 16342816 TI - Heart failure in African Americans. New research demands attention to race. PMID- 16342817 TI - Assessing the athlete at heart. Cardiovascular screening in sports. PMID- 16342818 TI - In-office cardiac arrest. A systematic approach to quality care. PMID- 16342819 TI - Getting pumped. Continuous insulin infusion therapy. PMID- 16342820 TI - Only good press in 2005. Patients gained access, but NPs faced increased opposition. PMID- 16342821 TI - Annual legislative update. State initiatives in 2005. PMID- 16342822 TI - Slice at life. Teens who cut, burn and beat themselves to dull inner pain. PMID- 16342823 TI - Is the practice doctorate needed? PMID- 16342824 TI - AFP benefits from medical editing clerkship. PMID- 16342825 TI - Tramadol for acute pain: a review of the evidence. PMID- 16342826 TI - Encounter form for patients with acute knee injuries. PMID- 16342827 TI - Case report: expanding the differential diagnosis of intractable cough. PMID- 16342828 TI - Time of hope for the eventual elimination of meningococcal strains A, C, Y, and W 135 in the United States. PMID- 16342829 TI - Trial of labor after cesarean delivery: the making of a recommendation. PMID- 16342830 TI - Who will have health insurance in 2025? AB - If current trends continue, U.S. health insurance costs will consume the average household's annual income by 2025. As health care becomes unaffordable for most people in the United States, it will be necessary to implement innovative models to move the system in a more equitable and sustainable direction. PMID- 16342831 TI - Diagnosing the cause of chest pain. AB - Chest pain presents a diagnostic challenge in outpatient family medicine. Noncardiac causes are common, but it is important not to overlook serious conditions such as an acute coronary syndrome, pulmonary embolism, or pneumonia. In addition to a thorough history and physical examination, most patients should have a chest radiograph and an electrocardiogram. Patients with chest pain that is predictably exertional, with electrocardiogram abnormalities, or with cardiac risk factors should be evaluated further with measurement of troponin levels and cardiac stress testing. Risk of pulmonary embolism can be determined with a simple prediction rule, and a D-dimer assay can help determine whether further evaluation with helical computed tomography or venous ultrasound is needed. Fever, egophony, and dullness to percussion suggest pneumonia, which can be confirmed with chest radiograph. Although some patients with chest pain have heart failure, this is unlikely in the absence of dyspnea; a brain natriuretic peptide level measurement can clarify the diagnosis. Pain reproducible by palpation is more likely to be musculoskeletal than ischemic. Chest pain also may be associated with panic disorder, for which patients can be screened with a two item questionnaire. Clinical prediction rules can help clarify many of these diagnoses. PMID- 16342832 TI - Complications of body piercing. AB - The trend of body piercing at sites other than the earlobe has grown in popularity in the past decade. The tongue, lips, nose, eyebrows, nipples, navel, and genitals may be pierced. Complications of body piercing include local and systemic infections, poor cosmesis, and foreign body rejection. Swelling and tooth fracture are common problems after tongue piercing. Minor infections, allergic contact dermatitis, keloid formation, and traumatic tearing may occur after piercing of the earlobe. "High" ear piercing through the ear cartilage is associated with more serious infections and disfigurement. Fluoroquinolone antibiotics are advised for treatment of auricular perichondritis because of their antipseudomonal activity. Many complications from piercing are body-site specific or related to the piercing technique used. Navel, nipple, and genital piercings often have prolonged healing times. Family physicians should be prepared to address complications of body piercing and provide accurate information to patients. PMID- 16342833 TI - Information from your family doctor. Body piercing: what you should know. PMID- 16342834 TI - Tick-borne relapsing fever. AB - Tick-borne relapsing fever is characterized by recurring fevers separated by afebrile periods and is accompanied by nonspecific constitutional symptoms. It occurs after a patient has been bitten by a tick infected with a Borrelia spirochete. The diagnosis of tick-borne relapsing fever requires an accurate characterization of the fever and a thorough medical, social, and travel history of the patient. Findings on physical examination are variable; abdominal pain, vomiting, and altered sensorium are the most common symptoms. Laboratory confirmation of tick-borne relapsing fever is made by detection of spirochetes in thin or thick blood smears obtained during a febrile episode. Treatment with a tetracycline or macrolide antibiotic is effective, and antibiotic resistance is rare. Patients treated for tick-borne relapsing fever should be monitored closely for Jarisch-Herxheimer reactions. Fatalities from tick-borne relapsing fever are rare in treated patients, as are subsequent Jarisch-Herxheimer reactions. Persons in endemic regions should avoid rodent- and tick-infested areas and use insect repellents and protective clothing to prevent tick bites. PMID- 16342835 TI - Information from your family doctor. Tick-borne relapsing fever: what you should know. PMID- 16342836 TI - Prevention of meningococcal disease. AB - Invasive disease caused by Neisseria meningitidis has an average annual incidence of one case per 100,000 in the United States. The disease can be rapidly fatal or result in severe neurologic and vascular sequelae despite antibiotic therapy. Antibiotic chemoprophylaxis with rifampin, ciprofloxacin, or ceftriaxone is required for household and other close contacts. Although the majority of cases of meningococcal disease are sporadic, outbreaks can occur, and vaccination of the affected population often is necessary. Serogroup B accounts for the highest incidence of disease in young infants but is not contained in any vaccine licensed in the United States. Adolescents and young adults 15 to 24 years of age have a higher incidence of disease and a higher fatality rate than other populations. Because 70 to 80 percent of these infections in the United States are caused by meningococcal serogroups C, Y, and W-135, which are contained in the tetravalent meningococcal vaccines, they are potentially preventable. The U.S. Food and Drug Administration recently approved a meningococcal conjugate vaccine containing serogroups A, C, Y, and W-135. This T-cell-dependent vaccine induces bactericidal antibody production and promotes immunologic memory that should result in a longer duration of immunity. The Advisory Committee on Immunization Practices recommends that this vaccine be given to 11- and 12-year old adolescents, to adolescents entering high school, and to college freshmen living in dormitories. The vaccine also may be given to persons 11 to 55 years of age who belong to certain high-risk groups. PMID- 16342837 TI - Management of the difficult patient. AB - All physicians must care for some patients who are perceived as difficult because of behavioral or emotional aspects that affect their care. Difficulties may be traced to patient, physician, or health care system factors. Patient factors include psychiatric disorders, personality disorders, and subclinical behavior traits. Physician factors include overwork, poor communication skills, low level of experience, and discomfort with uncertainty. Health care system factors include productivity pressures, changes in health care financing, fragmentation of visits, and the availability of outside information sources that challenge the physician's authority. Patients should be assessed carefully for untreated psychopathology. Physicians should seek professional care or support from peers. Specific communication techniques and greater patient involvement in the process of care may enhance the relationship. PMID- 16342838 TI - Screening for testicular cancer: recommendation statement. PMID- 16342839 TI - Rash during pregnancy. PMID- 16342840 TI - Ezetimibe/Simvastatin (vytorin) for hypercholesterolemia. PMID- 16342841 TI - Sensitivity and specificity of urinary nitrite for UTIs. PMID- 16342842 TI - Culturally competent family medicine: transforming clinical practice and ourselves. PMID- 16342843 TI - Pharmacologic cardioversion for atrial fibrillation and flutter. PMID- 16342844 TI - Duration of therapy for women with uncomplicated UTI. PMID- 16342846 TI - Information from your family doctor. Tuberculosis: what you should know. PMID- 16342845 TI - Management of active tuberculosis. AB - Although the overall incidence of tuberculosis has been declining in the United States, it remains an important public health concern, particularly among immigrants, homeless persons, and persons infected with human immunodeficiency virus. Patients who present with symptoms of active tuberculosis (e.g., cough, weight loss, or malaise with known exposure to the disease) should be evaluated. Three induced sputum samples for acid-fast bacillus smear and culture should be obtained from patients with findings of tuberculosis or suspicion for active disease. If the patient has manifestations of extrapulmonary tuberculosis, smears and cultures should be obtained from these sites. Most patients with active tuberculosis should be treated initially with isoniazid, rifampin, pyrazinamide, and ethambutol for eight weeks, followed by 18 weeks of treatment with isoniazid and rifampin if needed. Repeat cultures should be performed after the initial eight-week treatment. PMID- 16342847 TI - Plantar fasciitis: evidence-based review of diagnosis and therapy. AB - Plantar fasciitis causes heel pain in active as well as sedentary adults of all ages. The condition is more likely to occur in persons who are obese or in those who are on their feet most of the day. A diagnosis of plantar fasciitis is based on the patient's history and physical findings. The accuracy of radiologic studies in diagnosing plantar heel pain is unknown. Most interventions used to manage plantar fasciitis have not been studied adequately; however, shoe inserts, stretching exercises, steroid injection, and custom-made night splints may be beneficial. Extracorporeal shock wave therapy may effectively treat runners with chronic heel pain but is ineffective in other patients. Limited evidence suggests that casting or surgery may be beneficial when conservative measures fail. PMID- 16342848 TI - Information from your family doctor. Plantar fasciitis: what you should know. PMID- 16342849 TI - St. John's wort. AB - St. John's wort has been used to treat a variety of conditions. Several brands are standardized for content of hypericin and hyperforin, which are among the most researched active components of St. John's wort. St. John's wort has been found to be superior to placebo and equivalent to standard antidepressants for the treatment of mild to moderate depression. Studies of St. John's wort for the treatment of major depression have had conflicting results. St. John's wort is generally well tolerated, although it may potentially reduce the effectiveness of several pharmaceutical drugs. PMID- 16342850 TI - Klinefelter syndrome. AB - To complement the 2005 Annual Clinical Focus on medical genomics, AFP is publishing a series of short reviews on genetic syndromes. This series was designed to increase awareness of these diseases so that family physicians can recognize and diagnose children with these disorders and understand the kind of care they might require in the future. This review discusses Klinefelter syndrome. PMID- 16342851 TI - Cross-cultural medicine. AB - Cultural competency is an essential skill for family physicians because of increasing ethnic diversity among patient populations. Culture, the shared beliefs and attitudes of a group, shapes ideas of what constitutes illness and acceptable treatment. A cross-cultural interview should elicit the patient's perception of the illness and any alternative therapies he or she is undergoing as well as facilitate a mutually acceptable treatment plan. Patients should understand instructions from their physicians and be able to repeat them in their own words. To protect the patient's confidentiality, it is best to avoid using the patient's family and friends as interpreters. Potential cultural conflicts between a physician and patient include differing attitudes towards time, personal space, eye contact, body language, and even what is important in life. Latino, Asian, and black healing traditions are rich and culturally meaningful but can affect management of chronic medical and psychiatric conditions. Efforts directed toward instituting more culturally relevant health care enrich the physician-patient relationship and improve patient rapport, adherence, and outcomes. PMID- 16342852 TI - Treatment of constipation in older adults. AB - Constipation is a common complaint in older adults. Although constipation is not a physiologic consequence of normal aging, decreased mobility and other comorbid medical conditions may contribute to its increased prevalence in older adults. Functional constipation is diagnosed when no secondary causes can be identified, such as a medical condition or a medicine with a side effect profile that includes constipation. Empiric treatment may be tried initially for patients with functional constipation. Management of chronic constipation includes keeping a stool diary to record the nature of the bowel movements, counseling on bowel training, increasing fluid and dietary fiber intake, and increasing physical activity. There are a variety of over-the-counter and prescription laxatives available for the treatment of constipation. Fiber and laxatives increase stool frequency and improve symptoms of constipation. If constipation is refractory to medical treatment, further diagnostic evaluation may be warranted to assess for colonic transit time and anorectal dysfunction. Alternative treatment methods such as biofeedback and surgery may be considered for these patients. PMID- 16342853 TI - Information from your family doctor. Constipation: what you should know. PMID- 16342854 TI - Screening for HIV: recommendation statement. PMID- 16342855 TI - Counseling or antidepressants for treating depression? PMID- 16342856 TI - Widespread pruritic papulopustules after returning from Thailand. PMID- 16342857 TI - Nonmedical ultrasonography during pregnancy. PMID- 16342858 TI - [Medical imaging: beyond the frontiers]. PMID- 16342859 TI - [Cardiac imaging]. PMID- 16342860 TI - [Abdominal imaging]. PMID- 16342861 TI - [Gynecologic imaging]. PMID- 16342862 TI - [Head and neck imaging]. PMID- 16342863 TI - [Musculoskeletal imaging]. PMID- 16342864 TI - [Breast imaging]. PMID- 16342865 TI - [Chest imaging]. PMID- 16342866 TI - [Genitourinary imaging]. PMID- 16342867 TI - [Computers in radiology]. PMID- 16342868 TI - [Neuroradiology]. PMID- 16342869 TI - [Oncologic imaging]. PMID- 16342870 TI - [Pediatric imaging]. PMID- 16342871 TI - [Radiation protection]. PMID- 16342872 TI - [US imaging]. PMID- 16342873 TI - [MRI: current trends]. PMID- 16342874 TI - [Sonography]. PMID- 16342875 TI - [CT 25 years after a Nobel Price ... Thank you Mr. Hounsfield!]. PMID- 16342876 TI - [Nuclear medicine: fusion imaging]. PMID- 16342877 TI - [Interventional radiology: proven outcomes, current trends]. PMID- 16342878 TI - [PACS: web based technology]. PMID- 16342879 TI - [Digital imaging: current trends]. PMID- 16342880 TI - [Aneurysmal bone cyst of the ethmoid bone]. AB - Aneurysmal bone cysts is a benign, slow growing expansile lesion usually found in long bones or vertebrae. Only 5 cases of ABC of the ethmoid bone have been reported in the literature. Plain radiographs are not specific. CT and MRI are very useful to depict the extent of the lesion and demonstrate the presence of blood-fluid levels, which is a characteristic finding of ABC. We present one atypical case of ABC of the ethmoid bone in a 15-year-old female characterised by its extensive involvement and its rare location. PMID- 16342881 TI - [Transverse fracture of the sacrum: value of imaging. A case report]. AB - Transverse fractures of the sacrum with major displacement are rare and often misdiagnosed. A case of transverse fracture of the sacrum with cauda equina injury is reported. The diagnosis was not recognized initially. Conventional radiographs of the pelvis failed to demonstrate the fracture. True lateral sacral views and CT scan with reconstructions allowed analysis of the different sagittal fracture lines to facilitate surgical planning. These examinations should be considered in all patients with history of high energy trauma and clinical signs indicating lumbosacral injury. PMID- 16342882 TI - [A rare cause of hematuria: rupture of a renal artery aneurysm. A case report]. AB - The authors report a case of ruptured renal artery aneurysm in a 78 year old patient presenting with macroscopic hematuria. The rare occurrence of this entity and the value of imaging for diagnosis are discussed. PMID- 16342883 TI - [What is your diagnosis? Methanol poisoning]. PMID- 16342884 TI - [What is your diagnosis? Lobar dysmorphism of the kidney]. PMID- 16342885 TI - [What is your diagnosis? Leiomyosarcoma of the renal capsule]. PMID- 16342886 TI - [What is your diagnosis? Schwannoma of the trigeminal nerve]. PMID- 16342887 TI - [What is your diagnosis? Initial intramedullary sarcoidosis]. PMID- 16342888 TI - Developmental horizons: legacies and prospects in child and adolescent development. PMID- 16342889 TI - Looking back, for a change: a story of directions in child and adolescent development. PMID- 16342890 TI - Understanding Latino children and adolescents in the mainstream: placing culture at the center of developmental models. PMID- 16342891 TI - Essential role of culture in developmental psychology. PMID- 16342892 TI - Hindu worldview in the development of selfways: the "Atman" as the real self. PMID- 16342893 TI - Developing selves are meaning-making selves: recouping the social in self development. PMID- 16342894 TI - Children, adolescents, and the media: the molding of minds, bodies, and deeds. PMID- 16342895 TI - Intersection of social institutions with civic development. PMID- 16342896 TI - Family socialization: new moves and next steps. PMID- 16342897 TI - Friendship and development: putting the most human relationship in its place. PMID- 16342898 TI - Genes and experience in the development of executive attention and effortful control. PMID- 16342899 TI - Associations of emotion-related regulation with language skills, emotion knowledge, and academic outcomes. PMID- 16342900 TI - With eyes to the future: a brief history of cognitive development. PMID- 16342901 TI - CT for acute stage of closed head injury. AB - Brain damage after head injury can be classified by its time course. Primary damage that includes acute subdural hematoma (SDH), acute epidural hematoma (EDH), and intraaxial lesions that include contusions, diffuse axonal injury (DAI), and intracranial hemorrhage (ICH), occurs at the moment of impact and is thought to be irreversible. Secondary damage that includes herniations, diffuse cerebral swelling, and secondary infarction and hemorrhage, evolves hours or days after injury as a consequence of systemic or intracranial complications. The duration and severity of secondary damage influence outcome. Head injury management is focused on preventing, detecting, and correcting such secondary damage. CT has been widely used for the neuromonitoring of head trauma. CT is the gold standard for the detection of intracranial abnormalities and is a safe method for survey. While MRI is more sensitive and accurate in diagnosing cerebral pathology, CT is considered the most critical imaging technique for the management of closed head-injured patients in the acute stage. In this article, we review the imaging findings and literature of various lesions of closed head injury in the acute stage. PMID- 16342902 TI - Pelvic irradiation-induced eosinophilia is correlated to prognosis of cervical cancer patients and transient elevation of serum interleukin 5 level. AB - PURPOSE: To determine the significance of eosinophilia and serum interleukin 5 (IL-5) levels induced by radiotherapy in patients with gynecological cancer. MATERIALS AND METHODS: First, we examined changes in the number of eosinophils before and during radiotherapy in 145 patients with cervical cancer who underwent external radiotherapy and brachytherapy. Eosinophilia was defined as > or =10% eosinophils or > or =500 eosinophils/mm3. Next, we determined serum IL-5 levels prospectively in 20 patients, comprising 18 cases of cervical cancer, one of uterine corpus cancer, and one of vaginal cancer. RESULTS: Eosinophilia was seen in 88 patients (61%). Patients with advanced disease stage IV showed 32% eosinophila, lower than those of stage I (63%), II (69%), and III (65%) disease patients (p=0.03). The 5-year cause-specific survival rate was 76% for patients with eosinophilia (+) and 53% for patients without eosinophilia (p=0.0003). Thus, eosinophilia correlates to stage classification and a good prognosis. Of the 20 patients studied prospectively, the six with elevated serum IL-5 level at one week (6/20=30%) all showed eosinophilia (100%), whereas only six of the other 14 patients had eosinophilia (43%, p=0.04). CONCLUSION: Radiotherapy induced transient elevation of serum IL-5 levels and is related with eosinophilia, which is correlated to prognosis. PMID- 16342903 TI - Human comparative study of zinc and copper excretion via urine after administration of magnetic resonance imaging contrast agents. AB - PURPOSE: To evaluate the in-vivo pharmacokinetics of magnetic resonance imaging (MRI) contrast agents, the excretion of zinc and copper via urine was studied for three gadolinium (Gd) chelate complexes. MATERIALS AND METHODS: Urine samples were taken before, three hours, and six hours after intravenous administration of Gd-DTPA-BMA, Gd-DTPA, and Gd-DOTA at 0.2 ml/kg to five patients each who underwent contrast-enhanced MRI. Five patients who had non-contrast MRI were evaluated as controls. Urine was assayed for quantitative analysis of zinc and copper using atomic absorption analysis. RESULTS: Gd-DTPA-BMA caused the highest increase in zinc excretion among the three agents, 1,795 +/- 1,273 microg at 3 hours and 985 +/- 434 microg at 3 to 6 hours. Gd-DOTA did not cause a significant increase in zinc excretion, 75 +/- 39 microg at 3 hours and 78+/-65 microg at 3 to 6 hours. Gd-DTPA caused a moderate increase in zinc excretion, 665 +/- 240 microg at 3 hours and 378 +/- 173 microg at 3 to 6 hours. Excretion of copper did not show a significant difference among the three agents. CONCLUSION: Gd-DOTA was found to be the most kinetically inert among the three agents tested. The difference in zinc excretion among the MR contrast agents is possibly related to in-vivo transmetallation of the Gd chelate complexes correlated with variable stability of the contrast agents. The large amount of excess ligands contained in some MR contrast agents was also considered to be responsible for the increase of urinary zinc excretion. PMID- 16342904 TI - Radiotherapy for pelvic recurrence after radical hysterectomy for cervical cancer. AB - PURPOSE: To evaluate outcomes in patients treated with irradiation for recurrent cervical cancer after hysterectomy. METHODS: This is a retrospective chart review of 36 patients initially treated with a radical hysterectomy and lymph node dissection who developed a pelvic recurrence and were subsequently treated with irradiation. None received chemotherapy. RESULTS: The overall 5- and 10-year survivals were 74% and 50%, respectively. Ten patients developed recurrent disease after irradiation. The sites of failure in these 10 patients were pelvis only in two, pelvis and distant metastases in five, and distant metastases only in three. Severe complications occurred in four patients. One patient developed a hip fracture, one developed a bowel obstruction requiring a colostomy, and two developed fistulae. CONCLUSION: Pelvic irradiation and brachytherapy resulted in a 74% overall 5-year survival. Distant metastases were the most common site of failure. Systemic chemotherapy should be considered as adjunctive therapy for these patients. PMID- 16342905 TI - Weekly paclitaxel and carboplatin with concurrent radiation therapy in locally advanced non-small cell lung cancer: phase I study. AB - PURPOSE: The optimal dose of weekly paclitaxel in combination with carboplatin and concurrent radiation therapy (CRT) in patients with locally advanced non small cell lung cancer (NSCLC) is not well defined in Japan. The purpose of this study was to define the maximum-tolerated dose (MTD) of paclitaxel in this combination. MATERIALS AND METHODS: Treatment consisted of weekly paclitaxel plus carboplatin fixed AUC 2 and CRT for a total 60 Gy at 2 Gy/fraction/day 5 times weekly for 6 weeks. The dose of paclitaxel was 25 mg/m2 in the first cohort and escalated by 5 mg/m2 per cohort. RESULTS AND CONCLUSION: A total of 18 patients were enrolled and analyzed. The dose limiting toxicity of this study was considered to be pulmonary toxicity, and the MTD of it was determined to be 45 mg/m2 of paclitaxel in patients with NSCLC. Further evaluation of this regimen in a phase II trial is underway. PMID- 16342906 TI - Method of duplicating film using the CR system: evaluation of detectability in a simulated nodule. AB - PURPOSE: Since film processors used for screen-film systems have been decreasing recently, it is becoming difficult to develop duplicating film (Dup film) used conventionally. The purpose of this study was to evaluate the usefulness of the method of duplicating film using a computed radiography (CR) system. MATERIALS AND METHODS: The process of duplicating film using CR is 1) to eliminate energy accumulated on the imaging plate (IP) using white light, 2) to accumulate energy on the whole surface, and 3) to place the original film in piles. 4) After an exposure of white light, duplicated films can be obtained by CR system. In order to evaluate the reproducibiliy of our system, duplicated films were read by experienced observers and receiver operating characteristic (ROC) analysis was carried out. Observers read 50 images with a simulated nodule and 50 images without a simulated nodule. RESULTS: The average Az values were 0.94 for the original films, 0.91 for films duplicated using Dup film, and 0.90 for films duplicated using the CR system. When the two-tailed paired-T test was performed for each result, there were no statistically significant differences at p<0.05. CONCLUSION: The detectability of a simulated nodule for films duplicated using the CR system did not differ from the detectability of films duplicated using Dup film. This method may be a reasonable substitute for the conventional duplication system. PMID- 16342907 TI - Monte carlo dose calculations in conventional thorax fields for 60Co photons. AB - PURPOSE: Our purpose was to apply the MCNP4C Monte Carlo (MC) code for dose calculations in the thorax region and compare the results with those of measurements and a conventional treatment planning system (TPS). MATERIALS AND METHODS: We modeled a Theratron 780E 60Co unit and benchmarked our modeling with percent depth doses (PDDs), beam profiles, and output factors measured in a water phantom. For PDDs and beam profiles, the differences between measurements and MC calculations were less than 1% and 2%, respectively. We used an anatomic thorax phantom for evaluation of a conventional TPS and our MC calculation results. RESULTS: In comparing the results of calculations and measurements for our thorax geometries, the errors of conventional and MC methods were 20% and 2%, respectively. For the anterior mediastinal field and large thorax field the accuracy of the conventional method was acceptable, but for small fields of lateral thorax irradiation, the error of the conventional method was as high as to 20%. In all MC calculations, discrepancy from the measurements was less than 2%. CONCLUSION: Our results showed that the MCNP4C MC code could be used in dose calculations in treatment planning for 60Co photon irradiation. In addition, the application of the MC method for dose calculations in radiotherapy with 60Co photons was recommended. PMID- 16342908 TI - Evaluation of optimal scan duration and end time in cerebral CT perfusion study. AB - PURPOSE: The purpose of this study was to evaluate the optimal end time of scanning and the influence of varying the number of source images adopted for calculation of the parameter values in computerized tomography (CT) perfusion. MATERIALS AND METHODS: Nineteen CT perfusion studies in 14 patients with cerebrovascular disease were retrospectively analyzed. CT perfusion scanning was performed using continuous scans of 1 sec/rotation x60 sec with 5-mm-thick sections. To determine the appropriate end time of scanning, the time-density curves (TDCs) of the anterior cerebral artery (ACA), superior sagittal sinus (SSS), and basal ganglia were analyzed. The functional maps of cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT) were retrospectively generated from various numbers (30, 35, 40, 45, 50, 55, and 60) of source images. Defining the values calculated from the number of source images equal to the end time of the first pass as standard values, the percentage difference between the standard values and the values generated from various numbers of sources were evaluated. RESULTS: The TDCs of SSS showed the latest end time of the first pass (mean, 38.6+/-5.2 sec; range, 32 to 48 sec). Therefore, the values calculated from the number of source images equal to the end time of the first pass of SSS were defined as standard values. Increase and decrease of the number of source images for calculation resulted in increase of the percentage difference in every parameter value. The percentage differences ranged up to 49.6% for CBF, 48.0% for CBV, and 20.0% for MTT. CONCLUSIONS: Scanning until the end time of the first pass of SSS is necessary and sufficient for reliable measurement. Variable scan time based on the TDC of the SSS may be of better utility than use of fixed scan time. Further, the radiation dose could be minimized in many cases by reducing the scan time. PMID- 16342909 TI - Quantitative tissue blood flow evaluation of pancreatic tumor: comparison between xenon CT technique and perfusion CT technique based on deconvolution analysis. AB - PURPOSE: There has been one report that tissue blood flow (TBF) quantification with xenon CT was effective in predicting the therapeutic response to an anticancer drug in pancreatic cancer. The purpose of this study was to evaluate the correlation between the TBF of pancreatic tumors calculated with xenon CT and those with perfusion CT, in order to evaluate whether perfusion CT could replace xenon CT. MATERIALS AND METHODS: Nine patients with pathologically proved pancreatic tumors who underwent both xenon CT and perfusion CT were included. RESULTS: Quantitative TBF of pancreatic tumors measured by perfusion CT ranged from 22.1 to 196.2 ml/min/100 g (mean+/-SD, 52.6+/-54.8 ml/min/100 g). In contrast, those obtained by xenon CT ranged from 10.3 to 173.6 ml/min/100 g (mean+/-SD, 47.4+/-49.4 ml/min/100 g). There was a good linear correlation between xenon CT and perfusion CT (y=0.8537x+2.48, R2=0.895: p<0.05). CONCLUSION: The TBF of pancreatic tumors measured by xenon CT and perfusion CT techniques showed a close linear correlation. We can expect that perfusion CT based on the deconvolution algorithm may replace xenon CT to predict the effect of pancreatic tumor treatment with anticancer drugs. PMID- 16342910 TI - Hepatic hemangioma presenting atypical radiologic findings: a case report. AB - A 69-year-old woman was referred to our hospital due to a liver tumor that was incidentally noted on ultrasound (US). US revealed a pedunculated mass of 5 cm in diameter, with a heterogeneous echo pattern. On arterial phase dynamic contrast enhanced computed tomography (CT), a tiny enhancing dot in the upper aspect of the mass was seen; whereas, the main portion of the lesion appeared as hypoattenuating. The tumor was of low intensity on T1-weighted magnetic resonance (MR) images, and showed slightly heterogeneous high intensity on T2-weighted MR images. The most characteristic feature of the tumor was its exophytic appearance. On post-gadolinium hepatic arterial dominant-phase MR images, the tumor showed nodular enhancement centrally, with progressive spread of enhancement on later images. Angiography showed dilatation of the right posterior inferior branch of the hepatic artery and C-shaped opacification. Since we could not rule out malignancy for these nonspecific radiologic findings, a partial resection of the liver was carried out, resulting in a pathological diagnosis of hepatic hemangioma. This hemangioma had marked hyalinization and fibrosis, causing a heterogeneous appearance on MR images. The tumor presented an exophytic appearance, which caused some diagnostic confusion. PMID- 16342911 TI - Cerebral hyperperfusion in a patient with eclampsia with perfusion-weighted magnetic resonance imaging. AB - The patient was a 25-year-old Japanese woman with postpartum eclampsia, in whom magnetic resonance imaging (MRI) was performed ten hours after convulsion. The patient improved within three days. Diffusion-weighted images showed a high apparent diffusion coefficient (ADC), and perfusion-weighted images obtained by the multi-slice flow-sensitive alternating inversion recovery (FAIR) method showed hyperperfusion, which was suggestive of vasogenic edema, on the parietal and occipital subcortical white matter, corresponding with transient hyperintensities on T2-weighted images. MR angiography (MRA) revealed no signs of vasospasm. These findings suggested hyperperfusion-induced vasogenic edema without cerebrovascular spastic change in the early stage of eclampsia. The FAIR method is considered a useful technique for perfusion-weighted MRI, especially useful to evaluate the cerebral perfusion of emergency patients such as those with eclampsia. PMID- 16342912 TI - Use of the wallstent for symptomatic relief of malignant inferior vena cava obstructions. AB - PURPOSE: To report our experience with the Wallstent for the relief of congestive symptoms in patients with malignant inferior vena cava (IVC) obstructions. MATERIALS AND METHODS: We treated five patients with malignant obstructions of the IVC with a total of seven self-expandable stents. All obstructions were due to extrinsic compression; the procedures were indicated to relieve congestive symptoms in patients unresponsive to other therapies or in whom other treatments were contraindicated. Patients were evaluated for decrease in ascites and edema, weight loss, and patency, where available. RESULTS: All procedures were straightforward and obstructions could be successfully canalized. Inferior venacavograms obtained immediately after the procedure showed complete revascularization and disappearance of collateral circulation. Follow-up was carried out until death in four patients and until discharge in one. Of the five patients, four showed clinical improvement and one had no improvement. There was no recurrence of congestive symptoms. Except for one patient with partial migration of a stent, there were no important complications related to the procedure. CONCLUSION: Wallstents could be placed without complications in the IVC and were effective to ameliorate congestive symptoms in patients with malignant IVC obstructions. PMID- 16342913 TI - Ray tracing analysis of overlapping objects in refraction contrast imaging. AB - We simulated refraction contrast imaging in overlapping objects using the ray tracing method. The easiest case, in which two columnar objects (blood vessels) with a density of 1.0 [g/cm3], run at right angles in air, was calculated. For absorption, we performed simulation using the Snell law adapted to the object's boundary. A pair of bright and dark spot results from the interference of refracted X-rays where the blood vessels crossed. This has the possibility of increasing the visibility of the image. PMID- 16342914 TI - Research opportunities and challenges in 2005. AB - OBJECTIVES: Biomedical informatics practice and research have become so broad that I will comment on only three areas. These are the electronic health record; prospective population studies; and interactive publications. METHODS: Review of the literature, Internet resources and material and reports generated by internal and external research projects. RESULTS: Development of an electronic health record continues to be problematic in the U.S. A number of large-scale prospective cohort studies of normal populations are now actively occurring or in planning stage in a number of countries; these present special research opportunities in biomedical informatics. Although there are engineering challenges to the implementation of interactive publications, they have the potential to enhance understanding, learning, and memory of scientific and health concepts. CONCLUSIONS: Three areas of bio-medical informatics challenge us and offer interesting and worthy research opportunities. These include the electronic health record, prospective cohort studies, and interactive publications. PMID- 16342915 TI - Marfan syndrome--a diagnostic challenge caused by phenotypic and genetic heterogeneity. AB - OBJECTIVES: Marfan syndrome (MFS) is an autosomal dominant inherited connective tissue disorder caused by mutations in the fibrillin-1 (FBN1) gene with variable clinical manifestations in the cardiovascular, musculoskeletal and ocular systems. METHODS: Data of moleculor genetic analysis and a catalogue of clinical manifestations including aortic elastic parameters were mined in order to (i) assess aortic abnormality before and during medical treatment, and to (ii) identify novel correlations between the genotype and phenotype of the disease using hierarchical cluster analysis and logistic regression analysis. A score measure describing the similarity between a patient's clinical symptoms and a characteristic phenotype class was introduced. RESULTS: A probabilistic model for monitoring the loss of aortic elasticity was built on merely aortic parameters of 34 patients with classic MFS and 43 control subjects showing a sensitivity of 82% and a specificity of 96%. The clinical phenotypes of 100 individuals with classical or suspected MFS were clustered yielding four different phenotypic expressions. The highest correlation was found between FBN1 missense mutations, which manifested as ectopia lentis, skeletal major and skin minor criteria, and two out of four clustered phenotypes. The probability of the presence of a missense mutation in both phenotype classes is approximately 70%. CONCLUSIONS: Monitoring of aortic elastic properties during medical treatment may serve as additional criterion to indicate elective surgical interventions. Genotype phenotype correlation may contribute to anticipate the clinical consequences of specific FBN1 mutations more comprehensively and may be helpful to identify MFS patients at risk at on early stage of disease. PMID- 16342916 TI - A terminological and ontological analysis of the NCI Thesaurus. AB - OBJECTIVE: The National Cancer Institute Thesaurus is described by its authors as "a biomedical vocabulary that provides consistent, unambiguous codes and definitions for concepts used in cancer research" and which "exhibits ontology like properties in its construction and use". We performed a qualitative analysis of the Thesaurus in order to assess its conformity with principles of good practice in terminology and ontology design. MATERIALS AND METHODS: We used both the on-line browsable version of the Thesaurus and its OWL-representation (version 04.08b, released on August 2, 2004), measuring each in light of the requirements put forward in relevant ISO terminology standards and in light of ontological principles advanced in the recent literature. RESULTS: We found many mistakes and inconsistencies with respect to the term-formation principles used, the underlying knowledge representation system, and missing or inappropriately assigned verbal and formal definitions. CONCLUSION: Version 04.08b of the NCI Thesaurus suffers from the same broad range of problems that have been observed in other biomedical terminologies. For its further development, we recommend the use of a more principled approach that allows the Thesaurus to be tested not just for internal consistency but also for its degree of correspondence to that part of reality which it is designed to represent. PMID- 16342917 TI - A signal processing pipeline for noninvasive imaging of ventricular preexcitation. AB - OBJECTIVES: Noninvasive imaging of the cardiac activation sequence in humans could guide interventional curative treatment of cardiac arrhythmias by catheter ablation. Highly automated signal processing tools are desirable for clinical acceptance. The developed signal processing pipeline reduces user interactions to a minimum, which eases the operation by the staff in the catheter laboratory and increases the reproducibility of the results. METHODS: A previously described R peak detector was modified for automatic detection of all possible targets (beats) using the information of all leads in the ECG map. A direct method was applied for signal classification. The algorithm was tuned for distinguishing beats with an adenosine induced AV-nodal block from baseline morphology in Wolff Parkinson-White (WPW) patients. Furthermore, an automatic identification of the QRS-interval borders was implemented. RESULTS: The software was tested with data from eight patients having overt ventricular preexcitation. The R-peak detector captured all QRS-complexes with no false positive detection. The automatic classification was verified by demonstrating adenosine-induced prolongation of ventricular activation with statistical significance (p <0.001) in all patients. This also demonstrates the performance of the automatic detection of QRS-interval borders. Furthermore, all ectopic or paced beats were automatically separated from sinus rhythm. Computed activation maps are shown for one patient localizing the accessory pathway with an accuracy of 1 cm. CONCLUSIONS: The implemented signal processing pipeline is a powerful tool for selecting target beats for noninvasive activation imaging in WPW patients. It robustly identifies and classifies beats. The small beat to beat variations in the automatic QRS-interval detection indicate accurate identification of the time window of interest. PMID- 16342918 TI - Gene methylation data--a new challenge for bioinformaticians? AB - OBJECTIVES: Changes in the status of DNA methylation, known as epigenetic alterations, are among the most common molecular alterations in human neoplasia. For the first time, we reported on the analysis of fecal DNA from patients with CRC to determine the feasibility, sensitivity and specificity of this approach. We want to present basic information about DNA methylation analysis in the context of bioinformatics, the study design and several statistical experiences with gene methylation data. Additionally we outline chances and new research questions in the field of DNA methylation. METHODS: We present current approaches to DNA methylation analysis based on one reference study. Its study design and the statistical analysis is reflected in the context of biomarker development. Finally we outline perspectives and research questions for statisticians and bioinformaticians. RESULTS: Identification of at least three genes as potential DNA methylation-based tumor marker genes (SFRP2, SFRP5, PGR). CONCLUSIONS: DNA methylation analysis is a rising topic in molecular genetics. Gene methylation will push the extension of biobanks to include new types of genetic data. Study design and statistical methods for the detection of methylation biomarkers must be improved. For the purpose of establishing methylation analysis as a new diagnostic/prognostic tool the adaptation of several approaches has become a challenging field of research activity. PMID- 16342919 TI - Electronically signed documents in health care--analysis and assessment of data formats and transformation. AB - OBJECTIVES: Our objectives were to analyze and assess data formats for their suitability for conclusive and secure long-term archiving and to develop a concept for legally secure transformation of electronically signed documents that are not available in data formats appropriate for long-term archiving. METHODS: On the basis of literature review and Internet searches we developed general evaluation criteria to assess data formats with regard to their suitability for conclusive and secure long-term archiving. The assessment of data formats refers to format specifications and available literature. For the analyses of the transformation of signed documents we analyzed legal requirements on the basis of laws and ordinances as well as technical requirements by means of literature reviews, Internet searches and technical specifications. RESULTS: The following evaluation criteria are suited for this kind of assessment of data formats: transparency and standardization, stability, presentation and security. According to our assessment the following data formats are most suitable for conclusive and secure long-term archiving: PDF for formatted and unstructured text documents, XML for markup languages, TIFF for images in general, DICOM for medical images and S/MIME for the storage of e-mail. To transform electronically signed documents we propose an elementary procedure and universal basic model in form of an XML schema definition that includes the necessary legal and technical information. CONCLUSIONS: If electronic documents are to replace paper-based documents in patient records, they have to conform to the criteria for secure long-term archiving. The analyzed data formats are to be extended by mechanisms to guarantee the long-term security of electronic signatures. To transform large quantities of documents in a legally secure way, our basic model has to be extended for automated procedures. PMID- 16342920 TI - Current and future use of ICT for patient care and management in German acute hospitals--a comparison of the nursing and the hospital managers' perspectives. AB - OBJECTIVES: This study aimed at gaining comprehensive information on the current status of patient care and management applications used in German acute hospitals. Since the degree of ICT coverage in hospitals depends on the attitude of the key decision makers we also wanted to capture their plans and priorities and herewith try to predict future use. METHODS: We therefore conducted a nation wide survey including all acute hospitals in Germany in which two questionnaires were mailed to each hospital, one to the nursing managers, the other to the hospital managers. RESULTS: Six hundred hospitals participated in the survey which corresponds to an overall response rate of 27.6%. Accounting (84%) was found to be the most prevalent management module. Rostering was implemented in every second hospital. For clinical applications laboratory systems ranked first (69%). Ordering systems were used in nearly every second hospital. Nineteen percent of the hospitals reported employing an electronic patient record, 7% a nursing documentation system. Ranked by their priorities ordering systems hold the first position and care planning the last position. According to their plans, hospital managers, not nursing managers, intend to introduce nursing documentation. In contrast, nursing managers favor ordering and rostering for the near future. CONCLUSIONS: There is still a preponderance of management-oriented systems in German hospitals, yet clinical applications, in particular those supporting communications, will gain ground. The future of documentation systems is unclear, unless they not only provide statistical data for the management but support the clinical process properly. PMID- 16342921 TI - MorphoSaurus--design and evaluation of an interlingua-based, cross-language document retrieval engine for the medical domain. AB - OBJECTIVES: We propose an interlingua-based indexing approach to account for the particular challenges that arise in the design and implementation of cross language document retrieval systems for the medical domain. METHODS: Documents, as well as queries, are mapped to a language-independent conceptual layer on which retrieval operations are performed. We contrast this approach with the direct translation of German queries to English ones which, subsequently, are matched against English documents. RESULTS: We evaluate both approaches, interlingua-based and direct translation, on a large medical document collection, the OHSUMED corpus. A substantial benefit for interlingua-based document retrieval using German queries on English texts is found, which amounts to 93% of the (monolingual) English baseline. CONCLUSIONS: Most state-of-the-art cross language information retrieval systems translate user queries to the language(s) of the target documents. In contra-distinction to this approach, translating both documents and user queries into a language-independent, concept-like representation format is more beneficial to enhance cross-language retrieval performance. PMID- 16342922 TI - Creative and innovative statistics in clinical research and development. AB - OBJECTIVES: The aim of this paper is to show that even in a highly regulated area such as clinical research and development in pharmaceutical industry, there are needs and ample opportunities for statisticians and other medical informatics professionals to further creatively develop and implement methods in order to support the collection, analysis and interpretation of clinical data. METHODS: The recently published "Critical Path" initiative of the US Food and Drug Administration discusses the decline in new drug submissions in the last decade and illustrates potential causes in the present clinical development process. Areas where statisticians can and have begun to look for new innovative ways to overcome these shortcomings are presented and examples of such novel approaches that have been developed by statistical methodologists in the pharmaceutical industry together with statisticians in academia are given. RESULTS: In Early Development, i.e., in the first studies in man with a new compound, a combination of Bayesian methods and modeling approaches is particularly promising to increase the efficiency of decision making whereas in later phases (IIb and III) a marriage of modeling and classical frequentist approaches together with novel adaptive designs is expected help to chose the right dose regimen and to perform the trials more efficiently in reduced time. CONCLUSIONS: The combination of known statistical methods and thinking and the development of new approaches are in line with the present paradigm of "learning and confirming" in regulated clinical development while increasing the efficiency of both. PMID- 16342923 TI - Building multivariable regression models with continuous covariates in clinical epidemiology--with an emphasis on fractional polynomials. AB - OBJECTIVES: In fitting regression models, data analysts must often choose a model based on several candidate predictor variables which may influence the outcome. Most analysts either assume a linear relationship for continuous predictors, or categorize them and postulate step functions. By contrast, we propose to model possible non-linearity in the relationship between the outcome and several continuous predictors by estimating smooth functions of the predictors. We aim to demonstrate that a structured approach based on fractional polynomials can give a broadly satisfactory practical solution to the problem of simultaneously identifying a subset of 'important' predictors and determining the functional relationship for continuous predictors. METHODS: We discuss the background, and motivate and describe the multivariable fractional polynomial (MFP) approach to model selection from data which include continuous and categorical predictors. We compare our results with those from other approaches in examples. We present a small simulation study to compare the functional form of the relationship obtained by fitting fractional polynomials and splines to a single predictor variable. RESULTS: We illustrate the advantages of the MFP approach over standard techniques of model construction in two real example datasets analyzed with logistic and Cox regression models, respectively. In the simulation study, fractional polynomial models had lower mean square error and more realistic behaviour than comparable spline models. CONCLUSIONS: In many practical situations, the MFP approach can satisfy the aim of finding models that fit the data well and also are simple, interpretable and potentially transportable to other settings. PMID- 16342924 TI - An alternative proposal for "Mixed randomization" by Schulz and Grimes. AB - OBJECTIVE: Randomization is an important part of clinical trials. Using permuted block randomization for forcing equal group sizes potentially harms the unpredictability of treatment assignments. This can allow bias to creep into a trial. As an alternative, Schulz and Grimes suggest a "Mixed randomization" scheme which introduces more complexity to realize randomization. The objective of our research was to work out a model for randomization which is easier to handle than "Mixed randomization", with an equal level of performance in unpredictability and balance. METHODS: We analyzed a "Mixed randomization" procedure regarding the degree of unpredictability and balancing power and compared performance using permuted-block randomization with very large block size in a worst case scenario. Our work was done by the application of Blackwell Hodges model for evaluation of the unpredictability of treatment assignments. RESULTS: Regarding unpredictability, performance of permuted-block randomization with block size b = 36 was very similar to that of "Mixed randomization". Regarding balancing power it was more favourable than "Mixed randomization". CONCLUSION: Results of Schulz and Grimes are very important as they emphasized that mildly unequal sample sizes of therapy groups don't cause problems. But the suggested scheme of "Mixed randomization" to a large extent adds complexity and we do not believe that this proposal is very feasible. Basically, we rather recommend the use of only one restricted randomization procedure in the best way. This can be permuted-block randomization with optimum choice of a large block size. PMID- 16342925 TI - A simulation study using validated prognostic factors to assess expected long term survival. AB - OBJECTIVES: In chronic myeloid leukemia, after promising results in major cytogenetic remission (MCR), long-term survival data on imatinib treatment is of particular interest, especially in relation to former standard treatment based on interferon-alpha. However, data is still unavailable and due to high remission rates, most patients randomized to interferon-alpha in a clinical trial crossed over to imatinib. Therefore, to assess the expected long-term survival advantage with imatinib, a simulation study based on prognostic factors validated for interferon-alpha treatment was performed. METHODS: In interferon-alpha-treated patients with intermediate-risk and low-risk according to the established New CML score, survival probabilities of patients with MCR were significantly higher than those of patients without MCR. Three samples with simulated survival data for imatinib-treated intermediate-risk patients were constituted by randomly drawing varying percentages of their survival times from interferon-alpha-treated intermediate-risk patients with MCR and the remaining data from intermediate-risk patients without MCR. The same procedure was applied to low-risk patients. RESULTS: The 10-year survival probabilities of inter-feron-alpha-treated intermediate-risk and low-risk patients were 0.22 and 0.37. In the simulated samples, when 80%, 65%, and 50% of survival times were as favorable as for interferon-alpha-treated patients with MCR, respectively, the corresponding survival probabilities were 0.43 and 0.57, 0.36 and 0.49, and 0.30 and 0.42. CONCLUSIONS: In all simulation samples, increments of survival probabilities by imatinib were predicted, although survival probabilities of patients with MCR were assumed to be lower than with interferon-alpha. Prognosticated survival advantage with imatinib is backed by increasing observation time of imatinib treated patients in real studies. PMID- 16342926 TI - Genetic epidemiology in Germany--from biobanking to genetic statistics. AB - OBJECTIVES: Genetic epidemiology investigates the role of genetic factors and their interaction with environmental factors (in a broad meaning) for the occurrence of diseases in human populations. Its aim is to undestand the influence of genetics on the development of diseases, their course and the clinical implications, with the final goal to improve prevention, diagnostics and therapy. METHODS: Originally genetic epidemiology was understood as a specialized discipline with the main focus on family-based studies. The extraordinary development of genetics in the last decades--with respect of the understanding of the meaning of genes for human health, as well as by the availability of cost effective high throughput methods in the lab, has opened enormous opportunities to study genetic factors. Now, genetic epidemiology and genetic statistics have a much broader application. In addition, access to large samples of patients or from the population is needed. This can be realized via biobanks. RESULTS: Large biobanks with 500,000 or more patients or participants from the general population are being established or planned in the UK, Japan or the US. However, in Germany only two smaller activities are ongoing, KORA-gen in the south and POPGEN in the north. Possibilities to reach larger numbers, based on existing cohorts or disease networks are discussed. Ethical boundary conditions have to be taken into account, which seem to improve due to the Opinion of the German National Ethics Council on Biobanks for Research. Furthermore, the activities of the German centers for Genetic Epidemiological Methods (GEMs) as research and support units for genetic statistics and epidemiological methodology are described. CONCLUSIONS: Genetic epidemiology is based strongly on interdisciplinary collaboration and includes basics of genetics, elements of molecular biology to identify genes, population genetics, clinical medicine, and methodological disciplines as epidemiology, biostatistics and bioinformatics. In Germany the situation for this type of patient-based research has recently improved due to the National Genome Research Network (NGFN). PMID- 16342927 TI - A decision-support system for real-time risk assessment of airborne spread of the foot-and-mouth disease virus. AB - OBJECTIVES: [corrected] The application of epidemic models during the first days following the confirmation of a virus outbreak should significantly contribute to minimize its costs. Here we describe the first version of a decision-support system for the calculation of the airborne spread of a virus and its application to foot-and-mouth disease (FMD). The goal is to provide geographical maps depicting infection risk for various animal species to support the national health authorities. METHODS: The major tool of the decision-support system is a specific epidemic (or atmospheric) model: A so-called Gaussian dispersion model to calculate 3-dimensional virus plumes. Additional tools providing input data and visualizing the output are: A veterinary data base of geo-referenced premises, a geographical information system (GIS), and, as an external part running at the National Weather Service, a numerical weather prediction (NWP) model. To demonstrate the features of the decision-support system a pilot study in Styria, Austria, has been performed simulating an artificial FMD outbreak. RESULTS: One result of this simulation experiment is the determination of neighboring premises at which animals are at risk to be infected. Particular attention has been turned to cattle, sheep and swine. Using actual hourly NWP data from April 25, 2003, and a source of ten swine excreting a virus, cattle have been estimated to be at risk downwind 1,000-12,000 m, sheep 200-1,300 m, and swines 70-330 m. CONCLUSIONS: A system for real-time risk assessment of the airborne spread of a virus, applied to FMD, was introduced. Due to the forcing of the Gaussian dispersion model with NWP data, it is designed to run in both analysis and forecast mode. The system was applied for the first time during the Austrian real-time exercise on FMD, instructed by the European Union, in November 2004. PMID- 16342928 TI - Knowledge for medicine and health care--laudation at the occasion of the honorary doctorate bestowed to Donald A. B. Lindberg by UMIT, University for Health Sciences, Medical Informatics and Technology in Innsbruck, Tyrol, Austria. AB - Dr. Donald A. B. Lindberg, Director of the U.S. National Library of Medicine, received an honorary doctorate from UMIT, the University for Health Sciences, Medical Informatics and Technology in Innsbruck, Tyrol. The celebration took place on September 28, 2004 at an academic event during a conference of the Austrian, German, and Swiss Societies of Medical Informatics, GMDS2004. Dr. Lindberg has been a pioneer in the field of computers in health care from the early 1960s onwards. In 1984 he became the Director of the National Library of Medicine in Bethesda, the world's largest fully computerized biomedical library. Dr. Lindberg has been involved in the early activities of the International Medical Informatics Association (IMIA), among others being the chair of the Organizing Committee for MEDINFO 86 in Washington D.C. He was elected the first president of the American Medical Informatics Association (AMIA), and served as an editor of Methods of Information in Medicine. PMID- 16342929 TI - Diversity of function in the isocitrate lyase enzyme superfamily: the Dianthus caryophyllus petal death protein cleaves alpha-keto and alpha-hydroxycarboxylic acids. AB - The work described in this paper was carried out to define the chemical function a new member of the isocitrate lyase enzyme family derived from the flowering plant Dianthus caryophyllus. This protein (Swiss-Prot entry Q05957) is synthesized in the senescent flower petals and is named the "petal death protein" or "PDP". On the basis of an analysis of the structural contexts of sequence markers common to the C-C bond lyases of the isocitrate lyase/phosphoenolpyruvate mutase superfamily, a substrate screen that employed a (2R)-malate core structure was designed. Accordingly, stereochemically defined C(2)- and C(3)-substituted malates were synthesized and tested as substrates for PDP-catalyzed cleavage of the C(2)-C(3) bond. The screen identified (2R)-ethyl, (3S)-methylmalate, and oxaloacetate [likely to bind as the hydrate, C(2)(OH)(2) gem-diol] as the most active substrates (for each, k(cat)/K(m) = 2 x 10(4) M(-)(1) s(-)(1)). In contrast to the stringent substrate specificities previously observed for the Escherichia coli isocitrate and 2-methylisocitrate lyases, the PDP tolerated hydrogen, methyl, and to a much lesser extent acetate substituents at the C(3) position (S configuration only) and hydoxyl, methyl, ethyl, propyl, and to a much lesser extent isobutyl substituents at C(2) (R configuration only). It is hypothesized that PDP functions in oxalate production in Ca(2+) sequestering and/or in carbon scavenging from alpha-hydroxycarboxylate catabolites during the biochemical transition accompanying petal senescence. PMID- 16342930 TI - Crystal structure of the petal death protein from carnation flower. AB - Expression of the PSR132 protein from Dianthus caryophyllus (carnation, clover pink) is induced in response to ethylene production associated with petal senescence, and thus the protein is named petal death protein (PDP). Recent work has established that despite the annotation of PDP in sequence databases as carboxyphosphoenolpyruvate mutase, the enzyme is actually a C-C bond cleaving lyase exhibiting a broad substrate profile. The crystal structure of PDP has been determined at 2.7 A resolution, revealing a dimer-of-dimers oligomeric association. Consistent with sequence homology, the overall alpha/beta barrel fold of PDP is the same as that of other isocitrate lyase/PEP mutase superfamily members, including a swapped eighth helix within a dimer. Moreover, Mg(2+) binds in the active site of PDP with a coordination pattern similar to that seen in other superfamily members. A compound, covalently bound to the catalytic residue, Cys144, was interpreted as a thiohemiacetal adduct resulting from the reaction of glutaraldehyde used to cross-link the crystals. The Cys144-carrying flexible loop that gates access to the active site is in the closed conformation. Models of bound substrates and comparison with the closed conformation of isocitrate lyase and 2-methylisocitrate lyase revealed the structural basis for the broad substrate profile of PDP. PMID- 16342931 TI - The active site cysteine of ubiquitin-conjugating enzymes has a significantly elevated pKa: functional implications. AB - Ubiquitin-conjugating enzymes (E2s or Ubcs) are essential components in the ubiquitination apparatus. These enzymes accept ubiquitin from an E1 enzyme and then, usually with the aid of an E3 enzyme, donate the ubiquitin to the target protein. The function of E2 relies critically on the chemistry of its active site cysteine residue since this residue must form a thioester bond with the carboxyl terminus of ubiquitin. Despite the plethora of structural information that is available, there has been a notable dearth of information regarding the chemical basis of E2 function. Toward filling this large void in our understanding of E2 function, we have examined the pK(a) of the active site cysteine using a combination of experimental and theoretical approaches. We find, remarkably, that the pK(a) of the active site cysteine residue is elevated by approximately 2 pH units above that of a free cysteine. We have identified residues that contribute to the increase in this pK(a). On the basis of experimental values obtained with three different E2 proteins, we believe this to be a general and important characteristic of E2 protein chemistry. Sequence comparison suggests that the electrostatic environment is maintained not through strict residue conservation but through different combinations of residues near the active site. We propose that the elevated pK(a) is a regulatory mechanism that prevents the highly exposed cysteine residue in free E2 from reacting promiscuously with electron deficient chemical moieties in the cell. PMID- 16342932 TI - Role of Q52 in catalysis of decarboxylation and transamination in dialkylglycine decarboxylase. AB - Dialkylglycine decarboxylase (DGD) is a pyridoxal phosphate dependent enzyme that catalyzes both decarboxylation and transamination in its normal catalytic cycle. DGD uses stereoelectronic effects to control its unusual reaction specificity. X ray crystallographic structures of DGD suggest that Q52 is important in maintaining the substrate carboxylate in a stereoelectronically activated position. Here, the X-ray structures of the Q52A mutant and the wild type (WT) DGD-PMP enzymes are presented, as is the analysis of steady-state and half reaction kinetics of three Q52 mutants (Q52A, Q52I, and Q52E). As expected if stereoelectronic effects are important to catalysis, the steady-state rate of decarboxylation for all three mutants has decreased significantly compared to that of WT. Q52A exhibits an approximately 85-fold decrease in k(cat) relative to that of WT. The rate of the decarboxylation half-reaction decreases approximately 10(5)-fold in Q52I and approximately 10(4)-fold in Q52E compared to that of WT. Transamination half-reaction kinetics show that Q52A and Q52I have greatly reduced rates compared to that of WT and are seriously impaired in pyridoxamine phosphate (PMP) binding, with K(PMP) at least 50-100-fold greater than that of WT. The larger effect on the rate of l-alanine transamination than of pyruvate transamination in these mutants suggests that the rate decrease is the result of selective destabilization of the PMP form of the enzyme in these mutants. Q52E exhibits near-WT rates for transamination of both pyruvate and l-alanine. Substrate binding has been greatly weakened in Q52E with apparent dissociation constants at least 100-fold greater than that of WT. The rate of decarboxylation in Q52E allows the energetic contribution of stereoelectronic effects, DeltaG(stereoelectronic), to be estimated to be -7.3 kcal/mol for DGD. PMID- 16342933 TI - Role of the N-terminal extension of the (betaalpha)8-barrel enzyme indole-3 glycerol phosphate synthase for its fold, stability, and catalytic activity. AB - Indole-3-glycerol phosphate synthase (IGPS) catalyzes the fifth step in the biosynthesis of tryptophan. It belongs to the large and versatile family of (betaalpha)(8)-barrel enzymes but has an unusual N-terminal extension of about 40 residues. Limited proteolysis with trypsin of IGPS from both Sulfolobus solfataricus (sIGPS) and Thermotoga maritima (tIGPS) removes about 25 N-terminal residues and one of the two extra helices contained therein. To assess the role of the extension, the N-terminally truncated variants sIGPSDelta(1-26) and tIGPSDelta(1-25) were produced recombinantly in Escherichia coli, purified, and characterized in comparison to the wild-type enzymes. Both sIGPSDelta(1-26) and tIGPSDelta(1-25) have unchanged oligomerization states and turnover numbers. In contrast, their Michaelis constants for the substrate 1-(o-carboxyphenylamino)-1 deoxyribulose 5-phosphate are increased, and their resistance toward unfolding induced by heat and guanidinium chloride is decreased. sIGPSDelta(1-26) was crystallized, and its X-ray structure was solved at 2.8 A resolution. The comparison with the known structure of sIGPS reveals small differences that account for its reduced substrate affinity and protein stability. The structure of the core of sIGPSDelta(1-26) is, however, unchanged compared to sIGPS, explaining its retained catalytic activity and consistent with the idea that it evolved from the same ancestor as the phosphoribosyl anthranilate isomerase and the alpha-subunit of tryptophan synthase. These (betaalpha)(8)-barrel enzymes catalyze the reactions preceding and following IGPS in tryptophan biosynthesis but lack an N-terminal extension. PMID- 16342934 TI - Forster resonance energy transfer measurements are consistent with a helical bundle model for lipid-free apolipoprotein A-I. AB - Apolipoprotein (apo) A-I mutants were constructed for FRET studies to distinguish between two possible lipid-free conformers, a globular helix bundle and an elongated helical hairpin. Mutants containing a single Trp at position 50 were prepared by replacing Trps at positions 8, 72, and 108 with Phe (W@50). Two mutants were constructed from W@50 by incorporating Cys at Arg83 (W@50R83C) or Arg173 (W@50R173C) for attachment of the fluorescent probe AEDANS. Secondary structure of the mutants is very similar to wild type (wt) apo A-I, and fluorescence emission indicates that W50 is protected from solvent. Thermal stabilities of the AEDANS-labeled mutants are also similar to wt. These results indicate that no discernible changes occur in structure or stability as a result of mutations or labeling. The FRET data from W@50 to AEDANS are well-represented by a single distance distribution function with a distance of approximately 22 A for W@50R83C and approximately 19 A for W@50R173C. These distances are consistent with theoretical values calculated from a helical bundle model but not from a helical hairpin. A probability distance distribution function yields significantly small half-width values of 5.6 and 3.7 A, respectively, suggesting low conformational dynamics in both mutants. Differential scanning calorimetry (DSC) was performed on wt and a C-terminal deletion mutant, Delta(187-243), to obtain information on domain architecture. Contrary to expectations, both proteins unfold cooperatively. The results are consistent with the presence of a single folded domain within residues 1-186. These results support the presence of a discrete globular bundle conformation for lipid-free apo A-I. PMID- 16342935 TI - Tetraplex DNA transitions within the human c-myc promoter detected by multivariate curve resolution of fluorescence resonance energy transfer. AB - The nuclease hypersensitive element (NHE) III(I) of the c-myc promoter regulates the expression of oncogene c-myc and hence is an important anti-cancer target. Paranemic secondary structure formation within the promoter has been implicated in mechanistic regulation models. Here, it is shown that two monomeric tetraplexes form within the c-myc promoter, which coexist in solution. The development and application of a new experimental approach for detection of conformation transitions in nucleic acids [which exploits the sensitivity of fluorescence resonance energy transfer (FRET) for theoretical spectral resolution by multivariate curve resolution-alternating least-squares (MCR-ALS) method] has been used for this study. The pK(a) for tetraplex transitions are centered around 5.9 +/- 0.2 (between two intercalation topologies) and 6.8 +/- 0.1 (tetraplex to random coil). The presence of two tetraplexes has been further confirmed by S1 nuclease digestion. Finally, it is established that MCR-ALS analysis of FRET at different temperatures, pH, and salt concentrations allows resolution of pure species. Results are discussed in the light of recent observations implicating paranemic DNA motifs within the c-myc NHE in regulation of the oncogene. This method has several advantages over other methods vis-a-vis, high sensitivity and linear detection over a wide concentration range and, particularly, potential applications in intracellular probing. PMID- 16342936 TI - Calcium- and membrane-induced changes in the structure and dynamics of three helical hairpins in annexin B12. AB - Annexins are a family of soluble proteins that can undergo reversible Ca(2+) dependent interaction with the interfacial region of phospholipid membranes. The helical hairpins on the convex face of the crystal structure of soluble annexins are proposed to mediate binding to membranes, but the mechanism is not defined. For this study, we used a site-directed spin labeling (SDSL) experimental approach to investigate Ca(2+) and membrane-induced structural and dynamic changes that occurred in the helical hairpins encompassing three of the four D and E helices of annexin B12. Electron paramagnetic resonance (EPR) parameters were analyzed for the soluble and Ca(2+)-dependent membrane-bound states of the following nitroxide scans of annexin B12: a continuous 24-residue scan of the D and E helices in the third repeat (residues 219-242) and short scans encompassing the D-E loop regions of the first repeat (residues 68-74) and the fourth repeat (300-305). EPR mobility and accessibility parameters of most sites were similar when the protein was in solution or in the membrane-bound state, and both sets of data were consistent with the crystal structure of the protein. However, membrane induced changes in mobility and accessibility were observed in all three loop regions, with the most dramatic changes noted at sites corresponding to the highly conserved serine and glycine residues in the loops. EPR accessibility parameters clearly established that nitroxide side chains placed at these sites made direct contact with the bilayer. EPR mobility parameters showed that these sites were very mobile in solution, but immobilized on the EPR time scale in the membrane-bound state. Since the headgroup regions of bilayer phospholipids are relatively mobile in the absence of annexins, Ca(2+)-dependent binding of annexin B12 appears to form a complex in which the mobility of the D-E loop region of the protein and the headgroup region of the phospholipid are highly constrained. Possible biological consequences of annexin-induced restriction of membrane mobility are discussed. PMID- 16342937 TI - Structural conservation of mouse and rat zona pellucida glycoproteins. Probing the native rat zona pellucida proteome by mass spectrometry. AB - The mammalian zona pellucida is an egg extracellular matrix to which sperm bind. Mouse zonae are composed of three glycoproteins (ZP1, ZP2, and ZP3), while rat zonae contain four (ZP1, ZP2, ZP3, and ZP4/ZPB). Mouse sperm bind to zonae comprised solely of mouse ZP2 and ZP3. In this report, we show that rat sperm also bind to these zonae, indicating that ZP2 and ZP3 contain a "minimum structure(s)" to which rodent sperm can bind, and ZP1 and ZP4/ZPB are dispensable in these two rodents. These data are consistent with our mass spectrometric analysis of the native rat zona pellucida proteome (defined as the fraction of the total rat proteome to which the zonae glycoproteins contribute) demonstrating that the rat zonae glycoproteins share a high degree of conservation of structural features with respect to their mouse counterparts. The primary sequences of the rat zonae proteins have been deduced from cDNA. Each zona protein undergoes extensive co- and post-translational modification prior to its secretion and incorporation into an extracellular zona matrix. Each has a predicted N-terminal signal peptide that is cleaved off once protein translation begins and an anchoring C-terminal transmembrane domain from which the mature protein is released. Mass spectrometric analysis with a limited amount of native material allowed determination of the mature N-termini of rat ZP1 and ZP3, both of which are characterized by cyclization of glutamine to pyroglutamate; the N terminus of ZP2 was identified by Edman degradation. The mature C-termini of ZP1 and ZP3 end two amino acids upstream of a conserved dibasic residue that is part of, but distinct from, the consensus furin cleavage sequence, while the C terminus of ZP2 was not determined. Each zona protein contains a "zona domain" with eight conserved cysteine residues that is thought to play a role in the polymerization of the zona proteins into matrix filaments. Partial disulfide bond assignment indicates that the intramolecular disulfide patterns in rat ZP1, ZP2, and ZP3 are identical to those of their corresponding mouse counterparts. Last, nearly all potential N-glycosylation sites are occupied in the rat zonae glycoproteins (three of three for ZP1, six or seven of seven for ZP2, and four or five of six for ZP3). In comparison, potential O-glycosylation sites are numerous (59-83 Ser/Thr residues), but only two regions were observed to carry O-glycans in rat ZP3. PMID- 16342938 TI - A bivalent dissectional analysis of the high-affinity interactions between Cdc42 and the Cdc42/Rac interactive binding domains of signaling kinases in Candida albicans. AB - The small GTPase Cdc42, a member of the highly conserved Rho family of intracellular GTPases, communicates with downstream signaling proteins via high affinity interactions with the consensus Cdc42/Rac interactive binding (CRIB) polypeptide sequence. Previous biochemical and structural studies show that the CRIB motif itself is insufficient for high-affinity binding to Cdc42 but requires the sequence segment C-terminal to the CRIB motif for enhanced affinity. In this study, we have investigated the high-affinity (K(d) in units of nanomolar) associations of two highly homologous extended CRIB domains (eCRIBs) from the PAK kinases, Cla4 and Cst20, with Cdc42 from Candida albicans. (1)H-(15)N NMR heteronuclear NOE data of the eCRIB polypeptides in complex with Candida Cdc42 (CaCdc42) indicate that both eCRIB peptides have approximately two binding loci for CaCdc42. When each of the two eCRIB peptides is dissected into two fragments, the N-terminal fragments containing the minimal CRIB motif (mCRIB), mCla4 and mCst20, have relatively high binding affinities with dissociation constants (K(d)) of 4.2 and 0.43 microM, respectively. On the other hand, the C-terminal fragments (cCRIB), cCla4 and cCst20, exhibit significantly lower affinities for their binding to CaCdc42. The cCla4 peptide binds to CaCdc42 with a sub millimolar K(d) of 275 microM, and the cCst20 peptide shows an even lower binding affinity (K(d) = 1160 microM). Cross-titration experiments with the cognate fragments show that the binding affinity of cCst20 is enhanced approximately 5.5 fold (K(d) = 207 microM) in the presence of saturating amounts of mCst20, and vice versa. No such effect is observed for the binding of cCla4 and mCla4. These results suggest that the Cdc42-CRIB system can be represented by a "dual recognition" model for protein-protein interactions [Kleanthous, C., et al. (1998) Mol. Microbiol. 28, 227-233], following much the same mechanisms of multivalent molecular interactions [Song, J., and Ni, F. (1998) Biochem. Cell Biol. 76, 177-188; Mammen, M., et al. (1998) Angew Chem., Int. Ed. 37, 2754 2794]. The bivalent modeling of linked peptide fragments shows that the binding of eCla4 follows a simple additivity/avidity model, while binding of eCst20 appears to have a more complex mechanism involving cooperative effects. The differential binding mechanisms between closely related eCRIB polypeptides and CaCdc42 provide a new molecular basis for understanding kinase activation and for the design of antifungal agents targeting the large protein interaction interfaces engaged by the fungal GTPase. PMID- 16342939 TI - Prevention of MKK6-dependent activation by binding to p38alpha MAP kinase. AB - Inhibition of p38alpha MAP kinase is a potential approach for the treatment of inflammatory disorders. MKK6-dependent phosphorylation on the activation loop of p38alpha increases its catalytic activity and affinity for ATP. An inhibitor, BIRB796, binds at a site used by the purine moiety of ATP and extends into a "selectivity pocket", which is not used by ATP. It displaces the Asp168-Phe169 Gly170 motif at the start of the activation loop, promoting a "DFG-out" conformation. Some other inhibitors bind only in the purine site, with p38alpha remaining in a "DFG-in" conformation. We now demonstrate that selectivity pocket compounds prevent MKK6-dependent activation of p38alpha in addition to inhibiting catalysis by activated p38alpha. Inhibitors using only the purine site do not prevent MKK6-dependent activation. We present kinetic analyses of seven inhibitors, whose crystal structures as complexes with p38alpha have been determined. This work includes four new crystal structures and a novel assay to measure K(d) for nonactivated p38alpha. Selectivity pocket compounds associate with p38alpha over 30-fold more slowly than purine site compounds, apparently due to low abundance of the DFG-out conformation. At concentrations that inhibit cellular production of an inflammatory cytokine, TNFalpha, selectivity pocket compounds decrease levels of phosphorylated p38alpha and beta. Stabilization of a DFG-out conformation appears to interfere with recognition of p38alpha as a substrate by MKK6. ATP competes less effectively for prevention of activation than for inhibition of catalysis. By binding to a different conformation of the enzyme, compounds that prevent activation offer an alternative approach to modulation of p38alpha. PMID- 16342940 TI - Disulfide structure of the leucine-rich repeat C-terminal cap and C-terminal stalk region of Nogo-66 receptor. AB - Nogo-66 receptor (NgR1) is a leucine-rich repeat (LRR) protein that forms part of a signaling complex modulating axon regeneration. Previous studies have shown that the entire LRR region of NgR1, including the C-terminal cap of the LRR, LRRCT, is needed for ligand binding, and that the adjacent C-terminal region (CT stalk) of the NgR1 contributes to interaction with its coreceptors. To provide structure-based information for these interactions, we analyzed the disulfide structure of full-length NgR1. Our analysis revealed a novel disulfide structure in the C-terminal region of the NgR1, wherein the two Cys residues, Cys-335 and Cys-336, in the CT stalk are disulfide-linked to Cys-266 and Cys-309 in the LRRCT region: Cys-266 is linked to Cys-335, and Cys-309 to Cys-336. The other two Cys residues, Cys-264 and Cys-287, in the LRRCT region are disulfide-linked to each other. The analysis also showed that Cys-419 and Cys-429, in the CT stalk region, are linked to each other by a disulfide bond. Although published crystal structures of a recombinant fragment of NgR1 had revealed a disulfide linkage between Cys-266 and Cys-309 in the LRRCT region and we verified its presence in the corresponding fragment, this is artificially caused by the truncation of the protein, since this linkage was not detected in intact NgR1 or a slightly larger fragment containing Cys-335 and Cys-336. A structural model of the LRRCT with extended residues 311-344 from the CT stalk region is proposed, and its function in coreceptor binding is discussed. PMID- 16342941 TI - An elementary reaction step of the proton pump is revealed by mutation of tryptophan-164 to phenylalanine in cytochrome c oxidase from Paracoccus denitrificans. AB - Cytochrome c oxidase couples reduction of dioxygen to water to translocation of protons over the inner mitochondrial or bacterial membrane. A likely proton acceptor for pumped protons is the Delta-propionate of heme a(3), which may receive the proton via water molecules from a conserved glutamic acid (E278 in subunit I of the Paracoccus denitrificans enzyme) and which receives a hydrogen bond from a conserved tryptophan, W164. Here, W164 was mutated to phenylalanine (W164F) to further explore the role of the heme a(3) Delta-propionate in proton translocation. FTIR spectroscopy showed changes in vibrations possibly attributable to heme propionates, and the midpoint redox potential of heme a(3) decreased by approximately 50 mV. The reaction of the oxidized W164F enzyme with hydrogen peroxide yielded substantial amounts of the intermediate F' even at high pH, which suggests that the mutation rearranges the local electric field in the binuclear center that controls the peroxide reaction. The steady-state proton translocation stoichiometry of the W164F enzyme dropped to approximately 0.5 H(+)/e(-) in cells and reconstituted proteoliposomes. Time-resolved electrometric measurements showed that when the fully reduced W164F enzyme reacted with O(2), the membrane potential generated in the fast phase of this reaction was far too small to account either for full proton pumping or uptake of a substrate proton from the inside of the proteoliposomes. Time-resolved optical spectroscopy showed that this fast electrometric phase occurred with kinetics corresponding to the transition from state A to P(R), whereas the subsequent transition to the F state was strongly delayed. This is due to a delay of reprotonation of E278 via the D pathway, which was confirmed by observation of a slowed rate of Cu(A) oxidation and which explains the small amplitude of the fast charge transfer phase. Surprisingly, the W164F mutation thus mimics a weak block of the D-pathway, which is interpreted as an effect on the side chain isomerization of E278. The fast charge translocation may be due to transfer of a proton from E278 to a "pump site" above the heme groups and is likely to occur also in wild-type enzyme, though not distinguished earlier due to the high-amplitude membrane potential formation during the P(R)--> F transition. PMID- 16342942 TI - Computational studies of the farnesyltransferase ternary complex part I: substrate binding. AB - Farnesyltransferase (FTase) catalyzes the transfer of farnesyl from farnesyl diphosphate (FPP) to cysteine residues at or near the C-terminus of protein acceptors with a CaaX motif (a, aliphatic; X, Met). Farnesylation is a critical modification to many switch proteins involved in the extracellular signal transduction pathway, which facilitates their fixation on the cell membrane where the extracellular signal is processed. The malfunction caused by mutations in these proteins often causes uncontrolled cell reproduction and leads to tumor formation. FTase inhibitors have been extensively studied as potential anticancer agents in recent years, several of which have advanced to different phases of clinical trials. However, the mechanism of this biologically important enzyme has not been firmly established. Understanding how FTase recruits the FPP substrate is the first and foremost step toward further mechanistic investigations and the design of effective FTase inhibitors. Molecular dynamic simulations were carried out on the ternary structure of FTase complexed with the FPP substrate and an acetyl-capped tetrapeptide (acetyl-CVIM), which revealed that the FPP substrate maintains an inactive conformation and the binding of the diphosphate group can be largely attributed to residues R291beta, K164alpha, K294beta, and H248beta. The FPP substrate assumes an extended conformation in the binding site with restricted rotation of the backbone dihedral angles; however, it does not have a well-defined conformation when unbound in solution. This is evident from multinanosecond MD simulations of the FPP substrate in a vacuum and solution. Our conclusion is further supported by theoretical J coupling calculations. Our results on the FPP binding are in good agreement with previous experimental kinetic studies on FTase mutants. The hypothetical conformational activation of the FPP substrate is currently under investigation. PMID- 16342943 TI - Structural analysis of silanediols as transition-state-analogue inhibitors of the benchmark metalloprotease thermolysin. AB - Dialkylsilanediols have been found to be an effective functional group for the design of active-site-directed protease inhibitors, including aspartic (HIV protease) and metallo (ACE and thermolysin) proteases. The use of silanediols is predicated on its resemblance to the hydrated carbonyl transition-state structure of amide hydrolysis. This concept has been tested by replacing the presumed tetrahedral carbon of a thermolysin substrate with a silanediol group, resulting in an inhibitor with an inhibition constant K(i) = 40 nM. The structure of the silanediol bound to the active site of thermolysin was found to have a conformation very similar to that of a corresponding phosphonamidate inhibitor (K(i) = 10 nM). In both cases, a single oxygen is within bonding distance to the active-site zinc ion, mimicking the presumed tetrahedral transition state. There are binding differences that appear to be related to the presence or absence of protons on the oxygens attached to the silicon or phosphorus. This is the first crystal structure of an organosilane bound to the active site of a protease. PMID- 16342944 TI - Structural rationale for low-nanomolar binding of transition state mimics to a family GH3 beta-D-glucan glucohydrolase from barley. AB - The interactions of a transition state mimic anilinomethyl glucoimidazole (AmGlcIm), with a K(i) constant of 0.6 x 10(-)(9) M and a Gibbs free energy value of -53.5 kJ/mol, with a family GH3 beta-d-glucan glucohydrolase from barley have been analyzed crystallographically and by ab initio quantum mechanical modeling. AmGlcIm binds 3 times more tightly to the beta-d-glucan glucohydrolase than a previously investigated phenyl glucoimidazole. In the enzyme-AmGlcIm complex, an additional residue, Tyr253, and a water molecule positioned between subsites -1 and +1 are recruited for binding. Analyses of the two binary complexes reveal the following. (i) An intricate network exists in which hydrogen bonds between the enzyme's catalytic pocket residues Lys206, His207, Tyr253, Asp285, and Glu491 and the glucoimidazoles are shorter by 0.15-0.53 A, compared with distances of hydrogen bonds in the Michaelis complex. (ii) The "glucose" moiety of the glucoimidazoles adopts a (4)E conformation that is vital for the low-nanomolar binding. (iii) The N1 atoms of the glucoimidazoles are positioned nearly optimally for in-line protonation by the Oepsilon1 atom of the catalytic acid/base Glu491. (iv) The enzyme derives binding energies from both glycone and aglycone components of the glucoimidazoles. (iv) The prevalent libration motion of the two domains of the enzyme could play a significant role during induced fit closure in the active site. (v) Modeling based on the structural data predicts that protons could be positioned on the N1 atoms of the glucoimidazoles, and the catalytic acid/base Glu491 could carry an overall negative charge. (vi) The enzyme-AmGlcIm complex reveals the likely structure of an early transition state during hydrolysis. Finally, the high-resolution structures enabled us to define minimal structures of oligosaccharides attached to Asn221, Asn498, and Asn600 N glycosylation sites. PMID- 16342945 TI - Determinants in tRNA for activation of arginyl-tRNA synthetase: evidence that tRNA flexibility is required for the induced-fit mechanism. AB - Arginyl-tRNA synthetase (ArgRS) catalyzes formation of arginyl-adenylate in a tRNA-dependent reaction. Previous studies have revealed that conformational changes occur upon tRNA binding. In this study, we analyzed the sequence and structural features of tRNA that are essential to activate the catalytic center of mammalian arginyl-tRNA synthetase. Here, tRNA variants with different activator potential are presented. The three regions that are crucial for activation of ArgRS are the terminal adenosine, the D-loop, and the anticodon stem-loop of tRNA. The Add-1 N-terminal domain of ArgRS, which has the very unique property among aminoacyl-tRNA synthetases to interact with the D-loop in the corner of the convex side of tRNA, has an essential role in anchoring tRNA and participating in tRNA-induced amino acid activation. The results suggest that locking the acceptor extremity, the anticodon loop, and the D-loop of tRNA on the catalytic, anticodon-binding, and Add-1 domains of ArgRS also requires some flexibility of the tRNA molecule, provided by G:U base pairs, to achieve the productive conformation of the active site of the enzyme by induced fit. PMID- 16342946 TI - Potential of mean force calculation for the proton and hydride transfer reactions catalyzed by medium-chain acyl-CoA dehydrogenase: effect of mutations on enzyme catalysis. AB - Potential of mean force calculations have been performed on the wild-type medium chain acyl-CoA dehydrogenase (MCAD) and two of its mutant forms. Initial simulation and analysis of the active site of the enzyme reveal that an arginine residue (Arg256), conserved in the substrate-binding domain of this group of enzymes, exists in two alternate conformations, only one of which makes the enzyme active. This active conformation was used in subsequent computations of the enzymatic reactions. It is known that the catalytic alpha,beta dehydrogenation of fatty acyl-CoAs consists of two C-H bond dissociation processes: a proton abstraction and a hydride transfer. Energy profiles of the two reaction steps in the wild-type MCAD demonstrate that the reaction proceeds by a stepwise mechanism with a transient species. The activation barriers of the two steps differ by only approximately 2 kcal/mol, indicating that both may contribute to the rate-limiting process. Thus this may be a stepwise dissociation mechanism whose relative barriers can be tuned by suitable alterations of the substrate and/or enzyme. Analysis of the structures along the reaction path reveals that Arg256 plays a key role in maintaining the reaction center hydrogen bonding network involving the thioester carbonyl group, which stabilizes transition states as well as the intervening transient species. Mutation of this arginine residue to glutamine increases the activation barrier of the hydride transfer reaction by approximately 5 kcal/mol, and the present simulations predict a substantial loss of catalytic activity for this mutant. Structural analysis of this mutant reveals that the orientation of the thioester moiety of the substrate has been changed significantly as compared to that in the wild-type enzyme. In contrast, simulation of the active site of the Thr168Ala mutant shows no significant change in the relative orientation of the substrate and the cofactor in the active site; as a result, this mutation has very little effect on the overall reaction barrier, and this is consistent with the experimental data. This study demonstrates that significant insights into the catalytic mechanism can be obtained from simulation studies, and the results can be used to design novel mechanistic probes for the enzyme. PMID- 16342947 TI - Experimental validation of the docking orientation of Cdc25 with its Cdk2-CycA protein substrate. AB - Cdc25 phosphatases are key activators of the eukaryotic cell cycle and compelling anticancer targets because their overexpression has been associated with numerous cancers. However, drug discovery targeting these phosphatases has been hampered by the lack of structural information about how Cdc25s interact with their native protein substrates, the cyclin-dependent kinases. Herein, we predict a docked orientation for Cdc25B with its Cdk2-pTpY-CycA protein substrate by a rigid-body docking method and refine the docked models with full-scale molecular dynamics simulations and minimization. We validate the stable ensemble structure experimentally by a variety of in vitro and in vivo techniques. Specifically, we compare our model with a crystal structure of the substrate-trapping mutant of Cdc25B. We identify and validate in vivo a novel hot-spot residue on Cdc25B (Arg492) that plays a central role in protein substrate recognition. We identify a hot-spot residue on the substrate Cdk2 (Asp206) and confirm its interaction with hot-spot residues on Cdc25 using hot-spot swapping and double mutant cycles to derive interaction energies. Our experimentally validated model is consistent with previous studies of Cdk2 and its interaction partners and initiates the opportunity for drug discovery of inhibitors that target the remote binding sites of this protein-protein interaction. PMID- 16342948 TI - A slow, tight-binding inhibitor of the zinc-dependent deacetylase LpxC of lipid A biosynthesis with antibiotic activity comparable to ciprofloxacin. AB - The zinc-dependent enzyme LpxC catalyzes the deacetylation of UDP-3-O-acyl GlcNAc, the first committed step of lipid A biosynthesis. Lipid A is an essential component of the outer membranes of most Gram-negative bacteria, including Escherichia coli, Salmonella enterica, and Pseudomonas aeruginosa, making LpxC an attractive target for antibiotic design. The inhibition of LpxC by a novel N aroyl-l-threonine hydroxamic acid (CHIR-090) from a recent patent application (International Patent WO 2004/062601 A2 to Chiron and the University of Washington) is reported here. CHIR-090 possesses remarkable antibiotic activity against both E. coli and P. aeruginosa, comparable to that of ciprofloxacin. The biological activity of CHIR-090 is explained by its inhibition of diverse LpxC orthologues at low nanomolar concentrations, including that of Aquifex aeolicus, for which structural information is available. The inhibition of A. aeolicus LpxC by CHIR-090 occurs in two steps. The first step is rapid and reversible, with a K(i) of 1.0-1.7 nM, depending upon the method of assay. The second step involves the conversion of the EI complex with a half-life of about a minute to a tightly bound form. The second step is functionally irreversible but does not result in the covalent modification of the enzyme, as judged by electrospray ionization mass spectrometry. CHIR-090 is the first example of a slow, tight-binding inhibitor for LpxC and may be the prototype for a new generation of LpxC inhibitors with therapeutic applicability. PMID- 16342949 TI - Sanguinarine blocks cytokinesis in bacteria by inhibiting FtsZ assembly and bundling. AB - Bacterial diseases are among the leading causes of human death. The development of antibiotic resistance greatly contributes to the high mortality rate, and thus, the discovery of antibacterial drugs with novel mechanisms of action is needed. In this study, we found that sanguinarine, a benzophenanthridine alkaloid, strongly induced filamentation in both Gram-positive and Gram-negative bacteria and prevented bacterial cell division by inhibiting cytokinesis. Sanguinarine did not perturb the membrane structure in Escherichia coli. However, it perturbed the cytokinetic Z-ring formation in E. coli. In addition, sanguinarine strongly reduced the frequency of the occurrence of Z rings/micrometer of Bacillus subtilis length but did not alter the number of nucleoids/micrometer of cell length. The results suggested that sanguinarine inhibited cytokinesis in B. subtilis by inhibiting Z-ring formation without affecting nucleoid segregation. Sanguinarine inhibited the assembly of purified FtsZ and reduced the bundling of FtsZ protofilaments in vitro. Further, the interaction of sanguinarine to FtsZ was investigated using size-exclusion chromatography, an extrinsic fluorescent probe 1-anilinonaphthalene-8-sulfonic acid, and tryptophan fluorescence of mutated FtsZ (Y371W). Sanguinarine was found to bind to FtsZ with a dissociation constant of 18-30 microM. The results together show that sanguinarine inhibits bacterial division by perturbing FtsZ assembly dynamics in the Z ring and provide evidence in support of the hypothesis that the assembly and bundling of FtsZ play a critical role in bacterial cytokinesis. The results suggest that sanguinarine may be used as a lead compound to develop FtsZ-targeted antibacterial agents. PMID- 16342950 TI - Characterization of autocatalytic conversion of precursor BACE1 by heteronuclear NMR spectroscopy. AB - Accumulation of the cytotoxic 40- to 42-residue beta-amyloid peptide represents the primary pathological process in Alzheimer's disease (AD). BACE1 (beta-site APP cleaving enzyme 1) is responsible for the initial required step in the neuronal amyloidogenic processing of beta-amyloid precursor protein and is a major drug target for the therapeutic intervention of AD. In the present study, BACE1 is initially synthesized as an immature precursor protein containing part of the pre domain and the entire pro domain, and undergoes autocatalytic conversion to yield the well-folded mature BACE1 enzyme. To understand the mechanism of the conversion and the role of the pro domain, we monitored the autocatalytic conversion of BACE1 by heteronuclear NMR spectroscopy and used chemical shift perturbations as a probe to study the structural changes accompanying the autocatalytic conversion. NMR data revealed local conformational changes from a partially disordered to a well-folded conformation associated with the conversion. The conformational changes are largely concentrated in the NH(2) terminal lobe. Conversely, the active site conformations are conserved during the autocatalytic conversion. The precursor and mature BACE1 proteins were further characterized for their ability to interact with a substrate-based transition state BACE1 peptide inhibitor. The precursor BACE1 rapidly adopted the bound conformation in the presence of the inhibitor, which is identical to the bound conformation of the mature protein. The interaction of the inhibitor with both the precursor BACE1 and the fully processed BACE1 is in slow exchange on the NMR time scale, indicating a tight binding interaction. Overall, the NMR data demonstrated that the pro domain does not hinder inhibitor binding and may assist in the proper folding of the protein. The fully processed BACE1 represents a high quality well-folded protein which is highly stable over a long period of time, and is suitable for evaluation of inhibitor binding by NMR for drug intervention. PMID- 16342951 TI - Structural dynamics of the alpha-neurotoxin-acetylcholine-binding protein complex: hydrodynamic and fluorescence anisotropy decay analyses. AB - The three-fingered alpha-neurotoxins have played a pivotal role in elucidating the structure and function of the muscle-type and neuronal alpha7 nicotinic acetylcholine receptors (nAChRs). To advance our understanding of the alpha neurotoxin-nAChR interaction, we examined the flexibility of alpha-neurotoxin bound to the acetylcholine-binding protein (AChBP), which shares structural similarity and sequence identities with the extracellular domain of nAChRs. Because the crystal structure of five alpha-cobratoxin molecules bound to AChBP shows the toxins projecting radially like propeller "blades" from the perimeter of the donut-shaped AChBP, the toxin molecules should increase the frictional resistance and thereby alter the hydrodynamic properties of the complex. alpha Bungarotoxin binding had little effect on the frictional coefficients of AChBP measured by analytical ultracentrifugation, suggesting that the bound toxins are flexible. To support this conclusion, we measured the anisotropy decay of four site-specifically labeled alpha-cobratoxins (conjugated at positions Lys(23), Lys(35), Lys(49), and Lys(69)) bound to AChBP and free in solution and compared their anisotropy decay properties with fluorescently labeled cysteine mutants of AChBP. The results indicated that the core of the toxin molecule is relatively flexible when bound to AChBP. When hydrodynamic and anisotropy decay analyses are taken together, they establish that only one face of the second loop of the alpha neurotoxin is immobilized significantly by its binding. The results indicate that bound alpha-neurotoxin is not rigidly oriented on the surface of AChBP but rather exhibits segmental motion by virtue of flexibility in its fingerlike structure. PMID- 16342953 TI - Conformational changes of yeast plasma membrane H(+)-ATPase during activation by glucose: role of threonine-912 in the carboxy-terminal tail. AB - Yeast Pma1 H(+)-ATPase, which belongs to the P-type family of cation-transporting ATPases, is activated up to 10-fold by growth on glucose, and indirect evidence has linked the activation to Ser/Thr phosphorylation within the C-terminal tail. We have now used limited trypsinolysis to map glucose-induced conformational changes throughout the 100 kDa ATPase. In the wild-type enzyme, trypsin cleaves first at Lys-28 and Arg-73 in the extended N-terminal segment (sites T1 and T2); subsequent cleavages occur at Arg-271 between the A domain and M3 (site T3) and at Lys-749 or Lys-754 in the M6-M7 cytoplasmic loop (site T4). Activation by glucose leads to a striking increase in trypsin sensitivity. At the C-terminal end of the protein, the Arg- and Lys-rich tail is shielded from trypsin in membranes from glucose-starved cells (GS) but becomes accessible in membranes from glucose-metabolizing cells (GM). In the presence of orthovanadate, Lys-174 at the boundary between M2 and the A domain also becomes open to cleavage in GM but not GS samples (site T5). Significantly, this global conformational change can be suppressed by mutations at Thr-912, a consensus phosphorylation site near the C-terminus. Substitution by Ala at position 912 leads to a GS-like (trypsin resistant) state, while substitution by Asp leads to a GM-like (trypsin sensitive) state. Thus, the present results help to dissect the intramolecular movements that result in glucose activation. PMID- 16342952 TI - Partitioning conformational intermediates between competing refolding and aggregation pathways: insights into transthyretin amyloid disease. AB - Amyloid diseases are caused by the aberrant assembly of a protein in the extracellular space. Folded proteins are not amyloidogenic; however, the native state is generally in equilibrium with a minor population of unfolded or partially folded aggregation-competent conformers outside of the cell. Understanding how the partially unfolded conformers kinetically partition between the competing refolding and aggregation pathways provides insight into how misfolding, which occurs continuously, becomes pathogenic. Towards this end, we have previously studied the amyloidogenicity of transthyretin (TTR), a human beta sheet-rich homotetrameric protein that must undergo rate-limiting tetramer dissociation and partial monomer unfolding to misassemble into amyloid and other aggregates. We demonstrate herein that TTR homotetramers reassemble by an unusual monomer-dimer-trimer-tetramer (MDRT) pathway. Therefore, the rate of every step in the reassembly pathway is dependent on the concentration of folded TTR monomer. Partitioning soluble TTR monomers between the reassembly pathway and the aggregation pathway should therefore depend on the relative concentrations of aggregates and assembly intermediates. Aggregate clearance is envisioned to play an important role in the partitioning of protein in vivo, where partitioning to the aggregation pathway becomes increasingly favorable under conditions where the concentration of aggregates is increased because aggregate clearance is slow relative to the rate of aggregation. This shift from efficient to inefficient aggregate clearance could occur with aging, offering an explanation for the age associated nature of these neurodegenerative diseases. PMID- 16342954 TI - ATPase mechanism of Eg5 in the absence of microtubules: insight into microtubule activation and allosteric inhibition by monastrol. AB - The ATPase mechanism of kinesin superfamily members in the absence of microtubules remains largely uncharacterized. We have adopted a strategy to purify monomeric human Eg5 (HsKSP/Kinesin-5) in the nucleotide-free state (apoEg5) in order to perform a detailed transient state kinetic analysis. We have used steady-state and presteady-state kinetics to define the minimal ATPase mechanism for apoEg5 in the absence and presence of the Eg5-specific inhibitor, monastrol. ATP and ADP binding both occur via a two-step process with the isomerization of the collision complex limiting each forward reaction. ATP hydrolysis and phosphate product release are rapid steps in the mechanism, and the observed rate of these steps is limited by the relatively slow isomerization of the Eg5-ATP collision complex. A conformational change coupled to ADP release is the rate-limiting step in the pathway. We propose that the microtubule amplifies and accelerates the structural transitions needed to form the ATP hydrolysis competent state and for rapid ADP release, thus stimulating ATP turnover and increasing enzymatic efficiency. Monastrol appears to bind weakly to the Eg5-ATP collision complex, but after tight ATP binding, the affinity for monastrol increases, thus inhibiting the conformational change required for ADP product release. Taken together, we hypothesize that loop L5 of Eg5 undergoes an "open" to "closed" structural transition that correlates with the rearrangements of the switch-1 and switch-2 regions at the active site during the ATPase cycle. PMID- 16342955 TI - Definable equilibrium states in the folding of human prion protein. AB - The role of conformational intermediates in the conversion of prion protein from its normal cellular form (PrP(C)) to the disease-associated "scrapie" form (PrP(Sc)) remains unknown. To look for such intermediates in equilibrium conditions, we have examined the unfolding transitions of PrP(C), primarily using the chemical denaturant guanidine hydrochloride (GuHCl). When the protein conformation is assessed by NMR, there is a gradual shift of NMR signals in the regions between residues 125-146 and 186-196. The denaturant dependence of these shifts shows that in aqueous solution the native and locally unfolded conformations are both significantly populated. Following this shift, there is the major unfolding transition to generate a substantially unfolded population. However, analysis of NMR chemical shift and intensity changes shows that there is persistent structure in the molecule well beyond this major cooperative unfolding transition. Residual structure within this state is extensive and encompasses the majority of the secondary structure elements found in the native state of the protein. PMID- 16342956 TI - Implications of surface charge and curvature for the binding orientation of Thermomyces lanuginosus lipase on negatively charged or zwitterionic phospholipid vesicles as studied by ESR spectroscopy. AB - The triglyceride lipase (EC 3.1.1.3) Thermomyces lanuginosus lipase (TLL) binds with high affinity to unilamellar phospholipid vesicles that serve as a diluent interface for both lipase and substrate, but it displays interfacial activation on only small and negatively charged such vesicles [Cajal, Y., et al. (2000) Biochemistry 39, 413-423]. The productive-mode binding orientation of TLL at the lipid-water interface of small unilamellar vesicles (SUV) consisting of 1 palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG) was previously determined using electron spin resonance (ESR) spectroscopy in combination with site-directed spin-labeling [Hedin, E. M. K., et al. (2002) Biochemistry 41, 14185-14196]. In our investigation, we have studied the interfacial orientation of TLL when bound to large unilamellar vesicles (LUV) consisting of POPG, and bound to SUV consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC). Eleven single-cysteine TLL mutants were spin-labeled as previously described, and studied upon membrane binding using the water soluble spin relaxation agent chromium(III) oxalate (Crox). Furthermore, dansyl-labeled vesicles revealed the intermolecular fluorescence quenching efficiency between each spin-label positioned on TLL, and the lipid membrane. ESR exposure and fluorescence quenching data show that TLL associates closer to the negatively charged PG surface than the zwitterionic PC surface, and binds to both POPG LUV and POPC SUV predominantly through the concave backside of TLL opposite the active site, as revealed by the contact residues K74C-SL, R209C-SL, and T192C-SL. This orientation is significantly different compared to that on the POPG SUV, and might explain the differences in activation of the lipase. Evidently, both the charge and accessibility (curvature) of the vesicle surface determine the TLL orientation at the phospholipid interface. PMID- 16342957 TI - Assembly of tubulin by classic myelin basic protein isoforms and regulation by post-translational modification. AB - Myelin basic protein (MBP), a highly cationic protein that maintains the structure of the myelin sheath, associates with tubulin in vivo. The in vitro assembly of tubulin by MBP was examined here using several assays. The unmodified C1 component of 18.5 kDa bovine MBP (bC1) assembled tubulin into microtubules in a dose-dependent manner via filamentous intermediates, and was able simultaneously to promote the formation of microtubule bundles. The critical tubulin concentration in the presence of bC1 was 0.69 +/- 0.05 microM. The effects of post-translational modifications (such as deamidation and phosphorylation) were assayed by comparing the bC1-bC6 components of 18.5 kDa bovine MBP; an increasing level of modification enhanced the ability of MBP to assemble tubulin. The effects of charge reduction via deimination were examined using recombinant murine isoforms emulating the unmodified C1 and deiminated C8 isoforms of 18.5 kDa MBP; both rmC1 and rmC8 exhibited a comparable ability to assemble tubulin. The effects of alternate exon recombination of the classic MBP variants were tested using the recombinant murine 21.5, 17.22, and 14 kDa isoforms. The isoforms containing regions derived from exon II of the classic MBP gene, 21.5 and 17.22 kDa MBP, showed no substantial difference in the extent of tubulin polymerization and bundling when compared to those of 18.5 kDa MBP. The 14 kDa isoform and two terminal deletion mutants of rmC1 were able to induce microtubule polymerization, but not bundling, to the same degree as the longer proteins. Finally, bC1 was shown to disrupt and aggregate planar sheets of crystalline tubulin stabilized by paclitaxel, establishing that these structures are not suitable substrates for the formation of MBP cocrystals. PMID- 16342958 TI - Shotgun lipidomics identifies cardiolipin depletion in diabetic myocardium linking altered substrate utilization with mitochondrial dysfunction. AB - Diabetic cardiomyopathy is characterized by excessive utilization of fatty acid substrate, diminished glucose transport, and mitochondrial dysfunction. However, the chemical mechanisms linking altered substrate utilization to mitochondrial dysfunction are unknown. Herein, we use shotgun lipidomics and multidimensional mass spectrometry to identify dramatic decreases in the critical mitochondrial inner membrane lipid, cardiolipin, in diabetic murine myocardium (from 7.2 +/- 0.3 nmol/mg of protein in control hearts to 3.1 +/- 0.1 nmol/mg of protein in diabetic myocardium; p < 0.001, n = 7). Moreover, the direct metabolic precursor of cardiolipin, phosphatidylglycerol, was also substantially depleted (2.5 +/- 0.2 nmol/mg of protein in control hearts vs 1.3 +/- 0.1 nmol/mg of protein in diabetic myocardium; p < 0.001, n = 7). Similarly, glycerol 3-phosphate, necessary for the penultimate step in phosphatidylglycerol production, decreased by 58% in diabetic myocardium (from 4.9 +/- 0.9 to 2.2 +/- 0.3 nmol/mg of protein; n = 4). Since Barth's syndrome (a disorder of cardiolipin metabolism) induces mitochondrial dysfunction and cardiomyopathy, and since decreases in cardiolipin content precipitate mitochondrial dysfunction, these results provide a unifying hypothesis linking altered substrate utilization and metabolic flux in diabetic myocardium with altered lipid metabolism, cardiolipin depletion, mitochondrial dysfunction, and resultant hemodynamic compromise. PMID- 16342959 TI - Guanidine derivatives rescue the Arg418Ala mutation of Tritrichomonas foetus IMP dehydrogenase. AB - IMP dehydrogenase (IMPDH) catalyzes the oxidation of inosine 5'-monophosphate (IMP) to xanthosine 5'-monophosphate (XMP) and the reduction of NAD(+). The reaction involves formation of an E-XMP covalent intermediate; hydrolysis of the E-XMP intermediate is rate-limiting and requires the enzyme to adopt a closed conformation. Arg418 appears to act as the base that activates water for the hydrolysis reaction [Guillen-Schlippe, Y. V., and Hedstrom, L. (2005) Biochemistry 44, 11700-11707]. Deprotonation of Arg418 also stabilizes the closed conformation. Here we show that guanidine derivatives rescue the activity of the Arg418Ala variant. Amines and imidazole do not rescue. The rescue reaction appears to be saturable, with the values of K(R) ranging from 40 to 400 mM. The value of k(rescue) for the best rescue agents approaches the value of k(cat) for the reaction of the wild-type enzyme. Guanidine derivatives also rescue the activity of the Arg418Ala/Tyr419Phe variant. Multiple-inhibitor experiments suggest that the guanidine derivatives do not restore the equilibrium between open and closed conformations. Therefore, rescue agents must accelerate the hydrolysis of the E-XMP intermediate. The rate of the rescue reaction increases with an increase in pH, consistent with the hypothesis that the reaction involves neutral guanidine. A solvent D(2)O isotope effect is observed at low concentrations of the rescue agent, consistent with rate-limiting transfer of a proton from water. The value of k(cat) (rescue)/K(R)(base) correlates with the pK(a) of the guanidine derivative (Bronsted coefficient beta approximately 1). These results suggest that proton transfer from water to guanidine is almost complete in the transition state. PMID- 16342960 TI - Independent heterologous fibrillation of insulin and its B-chain peptide. AB - Insulin is very prone to form amyloid fibrils under slightly destabilizing conditions, and the B-chain region plays a critical role in the fibrillation. We show here that the isolated B-chain peptide of bovine insulin also forms fibrils at both acidic and neutral pH. When a mixture of insulin and the B-chain peptide was incubated at either acidic or neutral pH, the formation of fibrils was clearly separated into two phases, with the faster phase corresponding to the formation of homogeneous fibrils from the B-chain and the slower phase corresponding to homogeneous fibrillation of insulin. To further investigate the interaction (or lack thereof) between the two polypeptides, we examined the effects of cross-seeding. The results indicate that seeds of B-chain fibrils accelerate the fibrillation of insulin at pH 1.6 and inhibit the fibrillation at pH 7.5, but seeds of insulin fibrils have little effect on the fibrillation of the B-chain. We conclude that at pH 7.5 simultaneous independent homologous fibrillation occurs, but at low pH, heterologous fibrillation takes place, and with B-chain seeding of insulin, a unique conformation of fibrils is formed. Our results demonstrate that in the co-aggregation of closely related peptides each peptide species may undergo concurrent homogeneous or heterologous polymerization and that fibrils of one species may or may not seed fibrillation of the other. The results demonstrate the significant "species" barrier in amyloid fibril formation between fibrillation induced by different fibrils. A model for the fibrillation of the heterogeneous system of insulin and B-chain insulin is proposed. PMID- 16342961 TI - Thermodynamics of RNA duplexes with tandem mismatches containing a uracil-uracil pair flanked by C.G/G.C or G.C/A.U closing base pairs. AB - The thermodynamics governing the denaturation of RNA duplexes containing 8 bp and a central tandem mismatch or 10 bp were evaluated using UV absorbance melting curves. Each of the eight tandem mismatches that were examined had one U-U pair adjacent to another noncanonical base pair. They were examined in two different RNA duplex environments, one with the tandem mismatch closed by G.C base pairs and the other with G.C and A.U closing base pairs. The free energy increments (Delta Gdegrees(loop)) of the 2 x 2 loops were positive, and showed relatively small differences between the two closing base pair environments. Assuming temperature-independent enthalpy changes for the transitions, (Delta Gdegrees(loop)) for the 2 x 2 loops varied from 0.9 to 1.9 kcal/mol in 1 M Na(+) at 37 degrees C. Most values were within 0.8 kcal/mol of previously estimated values; however, a few sequences differed by 1.2-2.0 kcal/mol. Single strands employed to form the RNA duplexes exhibited small noncooperative absorbance increases with temperature or transitions indicative of partial self complementary duplexes. One strand formed a partial self-complementary duplex that was more stable than the tandem mismatch duplexes it formed. Transitions of the RNA duplexes were analyzed using equations that included the coupled equilibrium of self-complementary duplex and non-self-complementary duplex denaturation. The average heat capacity change (DeltaC(p)) associated with the transitions of two RNA duplexes was estimated by plotting DeltaH degrees and DeltaS degrees evaluated at different strand concentrations as a function of T(m) and ln T(m), respectively. The average DeltaC(p) was 70 +/- 5 cal K(-)(1) (mol of base pairs)(-)(1). Consideration of this heat capacity change reduced the free energy of formation at 37 degrees C of the 10 bp control RNA duplexes by 0.3-0.6 kcal/mol, which may increase Delta Gdegrees(loop) values by similar amounts. PMID- 16342962 TI - Probing heme propionate involvement in transmembrane proton transfer coupled to electron transfer in dihemic quinol:fumarate reductase by 13C-labeling and FTIR difference spectroscopy. AB - Quinol:fumarate reductase (QFR) is the terminal enzyme of anaerobic fumarate respiration. This membrane protein complex couples the oxidation of menaquinol to menaquinone to the reduction of fumarate to succinate. Although the diheme containing QFR from Wolinella succinogenes is known to catalyze an electroneutral process, its three-dimensional structure at 2.2 A resolution and the structural and functional characterization of variant enzymes revealed locations of the active sites that indicated electrogenic catalysis. A solution to this apparent controversy was proposed with the so-called "E-pathway hypothesis". According to this, transmembrane electron transfer via the heme groups is strictly coupled to a parallel, compensatory transfer of protons via a transiently established pathway, which is inactive in the oxidized state of the enzyme. Proposed constituents of the E-pathway are the side chain of Glu C180 and the ring C propionate of the distal heme. Previous experimental evidence strongly supports such a role of the former constituent. Here, we investigate a possible heme propionate involvement in redox-coupled proton transfer by a combination of specific (13)C-heme propionate labeling and Fourier transform infrared (FTIR) difference spectroscopy. The labeling was achieved by creating a W. succinogenes mutant that was auxotrophic for the heme-precursor 5-aminolevulinate and by providing [1-(13)C]-5-aminolevulinate to the medium. FTIR difference spectroscopy revealed a variation on characteristic heme propionate vibrations in the mid infrared range upon redox changes of the distal heme. These results support a functional role of the distal heme ring C propionate in the context of the proposed E-pathway hypothesis of coupled transmembrane electron and proton transfer. PMID- 16342963 TI - Spectroscopic and biochemical characterization of heme binding to yeast Dap1p and mouse PGRMC1p. AB - Yeast damage-associated response protein (Dap1p) and mouse progesterone receptor membrane component-1 protein (mPGRMC1p) belong to a highly conserved class of putative membrane-associated progesterone binding proteins (MAPR), with Dap1p and inner zone antigen (IZA), the rat homologue of mPGRMC1p, recently being reported to bind heme. While primary structure analysis reveals similarities to the cytochrome b(5) motif, neither of the two axial histidines responsible for ligation to the heme is present in any of the MAPR proteins. In this paper, EPR, MCD, CD, UV-vis, and general biochemical methods have been used to characterize the nature of heme binding in both Dap1p and a His-tagged, membrane anchor truncated mPGRMC1p. As isolated, Dap1p is a tetramer which can be converted to a dimer upon addition of 150 mM salt. The heme is noncovalently attached, with a maximal, in vitro, heme loading of approximately 30%, for both proteins. CD and fluorescence spectroscopies indicate a well-ordered structure, suggesting the low level of heme loading is probably not due to improperly folded protein. EPR confirmed a five-coordinate, high-spin, ferric resting state for both proteins, indicating one axial amino acid ligand, in contrast to the six-coordinate, low spin, ferric state of cytochrome b(5). The MCD spectrum confirmed this conclusion for Dap1p and indicated the axial ligand is most likely a tyrosine and not a histidine, or a cysteine; however, an aspartic acid residue could not be conclusively ruled out. Potential axial ligands, which are conserved in all MAPRs, were mutated (Y78F, D118A, and Y138F) and purified to homogeneity. The Y78F and D118A mutants were found to bind heme; however, Y138F did not. This result is consistent with the MCD data and indicates that Tyr138 is most likely the axial ligand to the heme in Dap1p. PMID- 16342964 TI - The reaction of LipB, the octanoyl-[acyl carrier protein]:protein N octanoyltransferase of lipoic acid synthesis, proceeds through an acyl-enzyme intermediate. AB - The lipB gene of Escherichia coli encodes an enzyme (LipB) that transfers the octanoyl moiety of octanoyl-acyl carrier protein (octanoyl-ACP) to the lipoyl domains of the 2-oxo acid dehydrogenases and the H subunit of glycine cleavage enzyme. We report that the LipB reaction proceeds through an acyl-enzyme intermediate in which the octanoyl moiety forms a thioester bond with the thiol of residue C169. The intermediate was catalytically competent in that the octanoyl group of the purified octanoylated LipB was transferred either to an 87 residue lipoyl domain derived from E. coli pyruvate dehydrogenase or to ACP (in the reversal of the physiological reaction). The octanoylated LipB linkage was cleaved by thiol reagents and by neutral hydroxylamine, strongly suggesting a thioester bond. Separation and mass spectral analyses of the peptides of the unmodified and octanoylated proteins showed that each of the assigned peptides of the two proteins had identical masses, indicating that none of these peptides were octanoylated. However, the one major peptide that we failed to recover was that predicted to contain all three LipB cysteine residues. These three cysteine residues were therefore targeted for site-directed mutagenesis and only C169 was found to be essential for LipB function in vivo. The C169S protein had no detectable activity whereas the C169A protein retained trace activity. Surprisingly, both proteins lacking C169 formed an octanoyl-LipB species, although neither was catalytically competent. The octanoyl-LipB species formed by the C169S protein was resistant to neutral hydroxylamine treatment, consistent with formation of an ester linkage to the serine hydroxyl group. The octanoyl C169A LipB species was probably acylated at C147. LipB species that lacked all three cysteine residues also formed a catalytically incompetent octanoyl adduct, indicating the presence of a reactive side chain other than a cysteine thiol that lies adjacent to the active site. PMID- 16342965 TI - Kernel energy method: application to DNA. AB - The kernel energy method (KEM) has been used in three recent papers (1-3) to calculate the quantum mechanical ab inito molecular energy of peptides and the protein insulin. It was found to have good accuracy. The computational difficulty of representing a molecule increases only modestly with the number of atoms. The calculations are simplified by adopting the approximation that a full biological molecule can be represented by smaller "kernels" of atoms. In this paper, the accuracy of the KEM is tested in the application to DNA, whose basic kernels, chemical bonding, and overall molecular structure are quite different from peptides and proteins. The basic kernel in the case of peptides and proteins is an amino acid. The basic kernel in the case of DNA is a nucleotide consisting of a phosphate-sugar-base. The molecular energy is calculated for all three basic types of DNA, i.e., B, A, and Z configurations of DNA. The results give an accuracy that is comparable to that achieved with peptides and proteins. Thus, the KEM is found to be applicable to major types of biological molecules. PMID- 16342966 TI - Kinetic and chemical mechanisms of the fabG-encoded Streptococcus pneumoniae beta ketoacyl-ACP reductase. AB - Beta-ketoacyl-acyl carrier protein reductase (KACPR) catalyzes the NADPH dependent reduction of beta-ketoacyl-acyl carrier protein (AcAc-ACP) to generate (3S)-beta-hydroxyacyl-ACP during the chain-elongation reaction of bacterial fatty acid biosynthesis. We report the evaluation of the kinetic and chemical mechanisms of KACPR using acetoacetyl-CoA (AcAc-CoA) as a substrate. Initial velocity, product inhibition, and deuterium kinetic isotope effect studies were consistent with a random bi-bi rapid-equilibrium kinetic mechanism of KACPR with formation of an enzyme-NADP(+)-AcAc-CoA dead-end complex. Plots of log V/K(NADPH) and log V/K(AcAc)(-)(CoA) indicated the presence of a single basic group (pK = 5.0-5.8) and a single acidic group (pK = 8.0-8.8) involved in catalysis, while the plot of log V vs pH indicated that at high pH an unprotonated form of the ternary enzyme complex was able to undergo catalysis. Significant and identical primary deuterium kinetic isotope effects were observed for V (2.6 +/- 0.4), V/K(NADPH) (2.6 +/- 0.1), and V/K(AcAc)(-)(CoA) (2.6 +/- 0.1) at pH 7.6, but all three values attenuated to values of near unity (1.1 +/- 0.03 or 0.91 +/- 0.02) at pH 10. Similarly, the large alpha-secondary deuterium kinetic isotope effect of 1.15 +/- 0.02 observed for [4R-(2)H]NADPH on V/K(AcAc)(-)(CoA) at pH 7.6 was reduced to a value of unity (1.00 +/- 0.04) at high pH. The complete analysis of the pH profiles and the solvent, primary, secondary, and multiple deuterium isotope effects were most consistent with a chemical mechanism of KACPR that is stepwise, wherein the hydride-transfer step is followed by protonation of the enolate intermediate. Estimations of the intrinsic primary and secondary deuterium isotope effects ((D)k = 2.7, (alpha)(-D)k = 1.16) and the correspondingly negligible commitment factors suggest a nearly full expression of the intrinsic isotope effects on (D)V/K and (alpha)(-D)V/K, and are consistent with a late transition state for the hydride transfer step. Conversely, the estimated intrinsic solvent effect ((D)2(O)k) of 5.3 was poorly expressed in the experimentally derived parameters (D)2(O)V/K and (D)2(O)V (both = 1.2 +/- 0.1), in agreement with the estimation that the catalytic commitment factor for proton transfer to the enolate intermediate is large. Such detailed knowledge of the chemical mechanism of KAPCR may now help guide the rational design of, or inform screening assay-design strategies for, potent inhibitors of this and related enzymes of the short chain dehydrogenase enzyme class. PMID- 16342967 TI - The Saccharomyces cerevisiae linker histone Hho1p, with two globular domains, can simultaneously bind to two four-way junction DNA molecules. AB - Saccharomyces cerevisiae encodes a single linker histone, Hho1p, with two globular domains. This raised the possibility that Hho1p could bind to two nucleosome cores simultaneously. To evaluate this idea, we studied the ability of a four-way junction, immobilized on the surface of a magnetic bead, to pull down a radiolabeled four-way junction in the presence of different Hho1 proteins. Four way junctions are known to bind to H1, presumably due to structure similarities to the DNA at the nucleosomal entry/exit point. We found a significant increase in the ability of full-length Hho1p to pull down radiolabeled four-way junction DNA under ionic conditions where both globular domains could bind. The binding was structure specific, since the use of double-stranded DNA, or a mutant Hho1p in which the second DNA binding site of globular domain 1 was abolished, resulted in a significant decrease in bridged binding. Additionally, bridged binding required a covalent attachment between the two globular domains, since factor Xa protease treatment of the complex formed by a modified Hho1p that contained a factor Xa cleavage site between the two globular domains resulted in a significant release of radiolabeled four-way junction. These findings demonstrated that the two globular domains independently associated with two different four-way junction molecules in a manner that required amino acid residues implicated in structure-specific binding in the nucleosome. We discuss the implication of these findings on the chromatin structure of yeast and propose a model where a single Hho1 protein binds to two serially adjacent nucleosomes. PMID- 16342969 TI - Magnetic-field-induced assemblies of cobalt nanoparticles. AB - Under the influence of a 0.05 T magnetic field, 15-nm diameter cobalt nanoparticles covered with surfactants in a colloidal solution assemble into highly constrained linear chains along the direction of the magnetic field. The magnetic-field-induced (MFI) chains become floppy after removal of the field, folding into three-dimensional (3D) coiled structures upon gentle agitation. The 3D structures are broken into smaller units with vigorous agitation. The nanoparticles redisperse into the solvent upon ultrasonic agitation. Optical microscopy and transmission electron microscopy (TEM) are used to characterize the morphologies of the nanoparticle assemblies at various stages of this reversible process. The hysteresis loops and zero-field cooled/field cooled (ZFC/FC) curves reveal the interparticle coupling in the assemblies. MFI assembly provides a powerful tool to manipulate magnetic nanoparticles. PMID- 16342968 TI - Analysis of interactions with mitochondrial mRNA using mutant forms of yeast NAD(+)-specific isocitrate dehydrogenase. AB - Yeast NAD(+)-specific isocitrate dehydrogenase (IDH) is an allosterically regulated tricarboxylic acid cycle enzyme that has been shown to bind specifically and with high affinity to 5'-untranslated regions of yeast mitochondrial mRNAs. The absence of IDH has been shown to result in reduced expression of mitochondrial translation products, leading to the suggestion that this macromolecular interaction may contribute to regulating rates of translation. The interaction with mitochondrial mRNAs also produces a dramatic inhibition of IDH catalytic activity that is specifically alleviated by AMP, the primary allosteric activator of IDH. Using mutant forms of IDH with defined catalytic or regulatory kinetic defects, we found that residue changes altering ligand binding in the catalytic site reduce the inhibitory effect of a transcript from the mitochondrial COX2 mRNA. In contrast, residue changes altering binding of allosteric regulators do not prevent inhibition by the COX2 RNA transcript but do prevent alleviation of inhibition by AMP. Results obtained using surface plasmon resonance methods suggest that the mRNA transcript may bind at the active site of IDH. Also, the presence of AMP has little effect on overall affinity but renders the binding of mRNA ineffective in catalytic inhibition of IDH. Finally, by expressing mutant forms of IDH in vivo, we determined that detrimental effects on levels of mitochondrial translation products correlate with a substantial reduction in catalytic activity. However, concomitant loss of IDH and of citrate synthase eliminates these effects, suggesting that any role of IDH in mitochondrial translation is indirect. PMID- 16342970 TI - Submerged microcontact printing (SmuCP): an unconventional printing technique of thiols using high aspect ratio, elastomeric stamps. AB - A technique for microcontact printing of thiols in liquid media is presented. Elastomeric poly(dimethyl siloxane) stamps are used to pattern gold surfaces with thiol-based self-assembled monolayers. The liquid (water in this case) has been used as an incompressible support and, advantageously, also acts as a medium in which alkylthiol ink molecules are poorly miscible. Consequently, we have been able to produce patterned thiol monolayers using stamps with aspect ratios unsuitable for conventional microcontact printing (i.e., 15:1) and present evidence to suggest that it is possible to use stamps with aspect ratios of up to 100:1. PMID- 16342971 TI - The AFM measured force required to rupture the dithiolate linkage of thioctic acid to gold is less than the rupture force of a simple gold-alkyl thiolate bond. AB - We show with atomic force microscopy that thioctic acid, a spatially constrained system with two sulfur linkages to gold, is less stable to tensile stress than a thiolate with a single attachment to gold. The force required to remove the dithiolate-linked thioctic acid was 0.31+/-0.13 nN, whereas the force required to remove a simple thiolate from the gold substrate was 1.05+/-0.29 nN. These results suggest that SAMs of densely packed or polypodal thiols may be substantially less stable under tensile stress than previously recognized and that the additional thiolate linkages may not only fail to increase the overall strength of attachment but could actually reduce it. PMID- 16342972 TI - Parallel alignment of carbon nanotubes induced with inorganic molecules. AB - In this work, a new strategy is presented to form ordered multiwalled carbon nanotube (MWNT) arrays. The MWNTs are aligned horizontally and parallel in (3 aminopropyl)triethoxysilane (3-APTES) sol films on the surface of mica and glassy carbon (GC). 3-APTES is ready to form charged rodlike micelles, which play a key role in fabricating orderly MWNT arrays. Moreover, we prepare MWNT arrays on the surfaces of solid electrodes and detect the electrochemical response of the microarray electrodes for the Fe(CN)(6)3-/Fe(CN)6(4-) couple. PMID- 16342973 TI - Control of carbon nanotube-surface interactions: the role of grafted polymers. AB - We have investigated the interactions between single-walled carbon nanotubes, coated with polymer chains end-grafted to the tubes, and planar surfaces. By proper functionalization of the grafted polymers' free ends, we show how to obtain an attractive interaction that can be used to immobilize the tube at a desired distance from the surface. We demonstrate how the strength and distance of the minimal interaction can be controlled by the proper choice of polymer chain length, surface coverage, and type of functional end-group. PMID- 16342974 TI - Grain boundary scars on spherical crystals. AB - We present an experimental system suitable for producing spherical crystals and for observing the distribution of lattice defects (disclinations and dislocations) on a significant fraction (50%) of the sphere. The introduction of fluorescently labeled particles enables us to determine the location and orientation of grain boundary scars. We find that the total number of scars and the number of excess dislocations per scar agree with theoretical predictions and that the geometrical centers of the scars are roughly positioned at the vertices of an icosahedron. PMID- 16342975 TI - Gold nanoparticle-cytochrome C complexes: the effect of nanoparticle ligand charge on protein structure. AB - We report the effect of nanoparticle ligand charge on the structure of a covalently, site-specifically linked protein. Au nanoparticles with positive, negative, and neutral ligands were appended to a specific cysteine, C102, of Saccharomyces cerevisiae cytochrome c. Conjugates were purified by HPLC or gel electrophoresis. Circular dichroism spectroscopy shows that changing the nanoparticle ligand dramatically influences the attached cytochrome c structure. The protein retains its structure with neutral ligands but denatures in the presence of charged species. This is rationalized by the electrostatic interaction of amino acids in the local vicinity of C102 with the endgroups of the ligand. PMID- 16342976 TI - Protein diffusion coefficients determined by macroscopic-gradient Rayleigh interferometry and dynamic light scattering. AB - Dynamic light scattering (DLS) is extensively used for measuring macromolecule diffusion coefficients. Contrary to classical techniques based on macroscopic concentration gradients, DLS probes microscopic fluctuations in concentration. DLS accuracy and its concordance with macroscopic-gradient techniques remains an outstanding important issue. We measured lysozyme diffusion coefficients in aqueous salt using both DLS and Rayleigh interferometry, a highly accurate macroscopic-gradient technique. The precision of our results is unprecedented. We find that our DLS values were systematically 2% higher than interferometry values. We believe that our interferometric measurements have produced the most accurate diffusion data ever reported for a protein, providing a new standard for quality control of DLS measurements. Furthermore, by interferometry, we have determined the whole diffusion coefficient matrix required for rigorously describing lysozyme-salt coupled diffusion. For the first time, we experimentally demonstrate that DLS does not provide the protein diffusion coefficient but one eigenvalue of the diffusion coefficient matrix. PMID- 16342977 TI - A qualitative theory of wimples in wetting films. AB - It has long been known that hydrodynamic pressures in a thin draining liquid film can cause inversion of the curvature of a drop surface as it approaches another surface, creating a so-called dimple. However, it was recently found that a different shape, dubbed a wimple, can be formed if a fluid drop, which is already in the field of repulsive surface forces, is abruptly pushed toward the wall. The drop shape might include a central region in which the film remains thin, surrounded by a ring of greater film thickness bounded at the outer edge by a barrier rim. Here we present a qualitative theory of the wimple formation. It is shown that this is mainly driven by the film hydrodynamics, and a qualitative criterion for the wimple/dimple transition is derived. PMID- 16342978 TI - New method for facile synthesis of amphiphilic thiol-stabilized ruthenium nanoparticles and their redox-active ruthenium nanocomposite. AB - The one-phase reduction of RuCl3 with lithium triethylborohydride as a reductant in tetrahydrofuran in the presence of 1-octanethiol, 1-octadecanethiol, 1,1' binaphthalene-2,2'-dithiol, or oligoethyleneoxythiol gave organic solvent- and water-soluble thiol-stabilized ruthenium nanoparticles. The oligoethyleneoxythiol stabilized ruthenium nanoparticles were soluble in both water and organic solvents. The ruthenium nanoparticles were stable in the solid state and did not aggregate in solution. Transmission electron microscope images of the ruthenium nanoparticles reveal small dispersed particles with a narrow size distribution. The ligand-exchange reaction of octadecanethiol-stabilized ruthenium nanoparticles (2.0 nm) with phenothiazine-linked decanethiol afforded redox active phenothiazine-functionalized ruthenium nanoparticles (1.9 nm) that showed a reversible redox peak at +0.51 V (vs Ag/0.1 M AgNO3) in the cyclic voltammogram. PMID- 16342979 TI - Reversible flocculation of silica across the phase boundary of poly(vinyl caprolactam) in aqueous solution. AB - The colloid stability of silica dispersions in water in the presence of poly(vinyl caprolactam) (PVCAP) has been studied below and above the lower consolute temperature (LCT) of its solutions. The dispersion sediments slowly without PVCAP in the temperature range studied (26-40 degrees C) or with PVCAP below the LCT ( approximately 30 degrees C). In contrast, with PVCAP above the LCT, rapid flocculation occurs at acid pH, with re-dispersal on cooling. Reversible flocculation is also obtained above the LCT by cycling the pH from alkaline to acid and back. The flocculation observed above the LCT may also be regarded as heterocoagulation between the silica particles and the aggregates of the polymer. PMID- 16342980 TI - Colloid vibration potential and ion vibration potential in a dilute suspension of spherical colloidal particles. AB - A general electroacoustic theory is presented for the macroscopic electric field in a dilute suspension of spherical colloidal particles in an electrolyte solution, which consists of the colloid vibration potential (CVP) and the ion vibration potential (IVP), induced by an oscillating pressure gradient field due to an applied sound wave. This is a unified theory that unites previous theories for CVP and those for IVP. Approximate analytic expressions are derived for CVP and IVP. The obtained IVP expression agrees with Debye's formula that is corrected by taking into account the force acting on the electrolyte ions as a result of the pressure gradient in the sound wave. The obtained CVP expression is correct to the first order of the particle zeta potential and applicable for arbitrary kappaalpha, where kappa is the Debye-Huckel parameter and alpha is the particle radius. It is found that an Onsager relation holds between CVP and dynamic electrophoretic mobility. It is also shown that the CVP from particles with very small kappaalpha approaches IVP; that is, in the limit of very small kappaalpha a particle behaves like an ion. PMID- 16342981 TI - Micelle-to-vesicle transition of an iron-chelating microbial surfactant, marinobactin E. AB - Small-angle neutron scattering (SANS) and dynamic light scattering (DLS) techniques have been applied to study the self-assembly processes of a microbially produced siderophore, marinobactin E (ME). ME is one of a series of marinobactins A-E that facilitate Fe(III) acquisition by the source bacterium through coordination of Fe(III) by the marinobactin headgroup. ME is a six-amino acid peptide amphiphile appended by palmitic acid (C16), and differs only in the nature of the fatty acid moiety from the other marinobactins. Apo-ME (uncoordinated ME) assembles to form micelles with an average diameter of 4.0 nm. Upon coordination of one equivalent of Fe(III), the mean micellar diameter of Fe(III)-ME shrinks to approximately 2.8 nm. However, in the presence of excess Fe(III), Fe(III)-ME undergoes a micelle-to-vesicle transition (MVT). At a small excess of Fe(III) over Fe(III)-ME (i.e., <1.2 Fe(III)/ME), a fraction of the Fe(III)-ME micelles rearrange into approximately 200 nm diameter unilamellar vesicles. At even greater Fe(III)/ME ratios (e.g., 2-3) multilamellar aggregates begin to emerge, consistent with either multilamellar vesicles or lamellar stacks. The MVT exhibited by ME may represent a unique mechanism by which marine bacteria may detect and sequester iron required for growth. PMID- 16342982 TI - Geometric constraints at the surfactant headgroup: effect on lipase activity in cationic reverse micelles. AB - The primary objective of the present article is to understand how the geometric constraints at the surfactant head affect the lipase activity in the reverse micellar interface. To resolve this issue, surfactants were designed and synthesized, and activity was measured in /water/isooctane/n-hexanol reverse micellar systems at z ([alcohol]/[surfactant])=5.6, pH 6.0 (20 mM phosphate), 25 degrees C across a varying range of W0 ([water]/[surfactant]) using p nitrophenylalkanoates as the substrate. It was observed that lipase activity increases from surfactants to with the increment in surface area per molecule (Amin) because of the substitution by the bulky tert-butyl group at the polar head. However, the activity was found to be similar for despite an enhancement in the hydrophilic moieties at the interface. This unchanged lipase activity is presumably due to the comparable surface area of to originating from the rigidity at the surfactant head. Noticeably, the enzyme activity improved from with the simultaneous increment of both the hydroxyl group and the flexibility of the headgroup whereas that for increased exclusively with the flexibility of the headgroup. The common parameter in both groups of surfactants and is the flexibility of the headgroup, which possibly enhance Amin and consequently the lipase activity. Thus, the geometric constraints at the surfactant headgroup play a crucial role in modulating the lipase activity profile probably because of the variation in interfacial area. PMID- 16342983 TI - Shrink-wrap vesicles. AB - We describe a simple approach to the controlled removal of molecules from the membrane of large unilamellar vesicles made of fatty acids. Such vesicles shrink dramatically upon mixing with micelles composed of a mixture of fatty acid and a phospholipid (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)), as fatty acid molecules leave the vesicle membrane and accumulate within the mixed micelles. Vesicle shrinkage was confirmed by dynamic light scattering, fluorescence recovery after photobleaching of labeled vesicles, and fluorescence resonance energy transfer between lipid dyes incorporated into the vesicle membrane. Most of the encapsulated impermeable solute is retained during shrinkage, becoming concentrated by a factor of at least 50-fold in the final small vesicles. This unprecedented combination of vesicle shrinkage with retention of contents allows for the preparation of small vesicles containing high solute concentrations, and may find applications in liposomal drug delivery. PMID- 16342984 TI - Donor-acceptor-promoted gelation of polyaromatic compounds. AB - Low molecular mass organogels are nonconventional polymeric structures in which a minute amount of low molecular weight compound can reversibly gelify the whole solution without forming covalent bonds between the monomers. In this article, we demonstrate that certain electron acceptors (taking dinitrobenzoates as model compounds) that are incapable of gelifying the solvent on their own can assemble as much as a 15-16-fold larger amount of polyaromatic hydrocarbons (PAHs) and form two-component donor-acceptor organogels in different solvents. At the core of the long-range order stand donor-acceptor pairs. We assess our claims by detailed 1H NMR, spectrophotometry, fluorescence, and time-resolved fluorescence methods. The thermodynamics of the gelation process is described on the basis of temperature dependent 1H NMR studies. We believe that, in this case, 1H NMR provides direct quantification of the dissolved concentrations of the different species and therefore provides a direct way to measure the enthalpy, entropy, and free energy associated with gel formation. PMID- 16342985 TI - Structure and dynamics in solvent-polarity-induced aggregates from a c60 fullerene-based dyad. AB - A novel methanofullerene dyad based on a hydrophobic (acceptor C60 moiety) hydrophilic (bridge with benzene and ester functionalities)-hydrophobic (donor didodecyloxybenzene) network is designed and synthesized. Electronic absorption spectral features revealed the molecule to exhibit a strong tendency to self aggregate in binary solvent mixtures at room temperature, where the dielectric constant exceeds a critical value, approximately 30. The dynamic structure factors of these spherical aggregates revealed stretched exponential decay with sizes varying between 110 and 250 nm with an increasing concentration, estimated from the dynamic light scattering experiments. However, a loss of shape selectivity of these aggregates was noted at lower water volume fractions in the binary solvent mixtures. The water-extracted spherical clusters were identified to be fractals with a dimension of 1.85, leading to diffusion-limited cluster aggregation as the mechanistic route for clusterization. PMID- 16342986 TI - Langmuir-Blodgett-Kuhn and self-assembled films of asymmetrically substituted poly(paraphenylene). AB - Asymmetrically substituted poly(paraphenylene) (PhPPP) with hydrophilic and hydrophobic side chains was investigated. The polymer behavior at the air-water interface was studied on the basis of surface pressure-area (pi-A) isotherms and compression/expansion hysteresis measurements. PhPPP can form stable monolayers with an area per repeat unit of A=0.20+/-0.02 nm2 and a collapse pressure in the range of pi=25 mN/m. Then, Langmuir-Blodgett-Kuhn (LBK) films of PhPPP were prepared by horizontally and vertically transferring the Langmuir monolayers onto hydrophilic solid substrates at pi=12 mN/m. Cross-section analysis of the AFM tapping-mode topography images of a single transferred monolayer reveals a thickness of d0=0.9+/-0.1 nm. Taking into account the obtained monolayer thickness, curve-fitting calculations of angular scan data of LB monolayers measured using surface plasmon resonance (SPR) spectroscopy lead to a value for the refractive index of n=1.78+/-0.02 at lambda=632.8 nm. Next, the spontaneous formation of a PhPPP monolayer by adsorption from solution was studied ex situ by atomic force microscopy and UV-vis spectroscopy and in situ by using SPR spectroscopy. Stable self-assembled monolayers of PhPPP can be formed on hydrophilic surfaces with a thickness similar to that of the monolayer obtained using the LB method. The characterization results confirmed the amphiphilic character and the self-assembly properties of PhPPP, as well as the possibility of preparing homogeneous monolayer and multilayer films. PMID- 16342987 TI - Refractive index of thin, aqueous films between hydrophobic surfaces studied using evanescent wave atomic force microscopy. AB - We have studied the refractive index of a thin aqueous film between microscopic hydrophobic surfaces using evanescent wave atomic force microscopy (EW-AFM). An evanescent wave, generated at a solid-liquid interface, is scattered by AFM tips or glass particles attached to AFM cantilevers. The scattering of this wave is used to determine the refractive index as a function of separation between these surfaces. Measurements were performed on surfaces that were rendered hydrophobic with octadecyltrichlorosilane, which produces solid-water contact angles in excess of 90 degrees. For AFM tips, the average refractive index in the thin film was always equal to that of water when the film was thicker than approximately 100 nm. At smaller separations, the refractive index was always greater than or equal to that of water. This is inconsistent with the formation of air or vapor films and consistent with a small amount of organic material between the surfaces. For colloidal spheres (R approximately 10 microm), we were not able to detect changes in the refractive index of the thin film between the sphere and plate. PMID- 16342988 TI - Molecular dynamics study of the influence of surfactant structure on surfactant facilitated spreading of droplets on solid surfaces. AB - The spreading of a partially wetting aqueous drop in air on a hydrophobic surface can be facilitated by the adsorption of surfactants from the drop phase onto the air/aqueous and aqueous/hydrophobic solid interfaces of the drop. At the contact line at which these interfaces meet, conventional surfactants with a linear alkyl hydrophobic chain attached to a polar group adsorb onto the surfaces, forming monolayers which remain distinct as they merge at the contact juncture. The adsorption causes a decrease in the interfacial tensions and reduction in the contact angle but the angle remains above zero so the drop is still nonwetting. Trisiloxane surfactants with a T-shaped geometry in which the hydrophobic group is composed of a trisiloxane oligomer with a polar group attached at the center of the chain can give rise to a zero contact angle at the contact line and complete wetting (superspreading). Experimental evidence suggests the adsorption of the T-shaped molecule, in addition to significantly decreasing the tensions of the interfaces (relative to the conventional surfactants), promotes the formation of a precursor film consisting of a surfactant bilayer at the contact line which facilitates the spreading. The aim of this study is to use molecular dynamics to examine if the T-shaped structure can promote spreading by the formation of a bilayer and to contrast this case with that of the linear chain surfactant where complex assembly does not occur. The simulation models the solvent as a monatomic liquid, the substrate as a particle lattice, and the surfactants as united atom structures, with all interactions given by Lennard-Jones potentials. We start with a base case in which the solvent partially wets a substrate comprised of a lattice of particles. We demonstrate that adsorbed T-shaped surfactant monolayers can, when the interaction between the solvent and the hydrophile particles is strong enough, assemble into a bilayer, allowing the drop to extend to a thin planar film. In the case of the flexible linear chain surfactant, there is no interaction between the monolayers on the two interfaces in the case of a strong hydrophile-solvent interaction and less coordination for a weaker interaction. In either case, the monolayers remain distinct, as the surfactant only marginally improves wetting. PMID- 16342989 TI - Odd-even variations in the wettability of n-alkanethiolate monolayers on gold by water and hexadecane: a molecular dynamics simulation study. AB - Molecular dynamics (MD) simulations were performed to investigate odd-even chain length dependencies in the wetting properties of self-assembled monolayers (SAMs) of n-alkanethiols [CH3(CH2)n-1SH] on gold by water and hexadecane. Experimentally, the contact angle of hexadecane on the SAMs depends on whether n is odd or even, while contact angles for water show no odd-even dependence. Our MD simulations of this system included a microscopic droplet of either 256 water molecules or 60 hexadecane molecules localized on an n-alkanethiolate SAM on gold with either an even or odd chain length. Contact angles calculated for these nanoscopic droplets were consistent with experimentally observed macroscopic trends in wettability, namely, that hexadecane is sensitive to structural differences between odd- and even-chained SAMs while water is not. Structural properties for the SAMs (including features such as chain tilt, chain twist, and terminal methyl group tilt) were calculated during the MD simulations and used to generate IR spectra of these films that compared favorably with experimental spectra. MD simulations of SAMs in contact with slabs of water and hexadecane revealed that the effects of these solvents on the structure of the SAM was restricted to the chain terminus and had no effect on the inner structure of the SAM. The density profiles for water and hexadecane on the SAMs were different in that water displayed a significant depletion in its density at the liquid/SAM interface from its bulk value, while no such depletion occurred for hexadecane. This difference in contact may explain the lack of an odd-even variation in the wetting characteristics of water on these surfaces, because the water molecules are positioned further away from the surface and, therefore, are not sensitive to the structural differences in the average orientations for the terminal methyl groups in odd- and even-chained SAMs. In contrast, the differences in the wetting properties of hexadecane on the odd- and even-chained SAMs may reflect the closer proximity of these molecules to the SAM surface and a resulting greater sensitivity to the differences in the terminal methyl group orientations in the SAMs. SAM-solvent interaction energies were calculated during the MD simulations, yielding interaction energies that differed on the even- and odd-chained surfaces by approximately 10% for hexadecane and negligibly for water, in accord with estimates using experimental wetting results. PMID- 16342990 TI - Interfacial tension and electrocapillary measurements of the room temperature ionic liquid/aqueous interface. AB - The surface and aqueous interfacial tensions for a series of water-immiscible room-temperature ionic liquids (RTILs) have been measured. The RTILs used in this study were based on 1-alkyl-3-methylimidazolium cations (Cnmim, n=6, 8, 10, and 12) and bis(perfluoromethylsulfonyl)imide (BMSI) and bis(perfluoroethylsulfonyl)imide (BETI) anions. It was found that the surface tensions of the RTILs increased with an increasing cation chain length similar to the behavior of n-alkanes. Interfacial tensions of the RTILs with aqueous solutions, however, were found to decrease with the cation chain length, which has been attributed to the increased surface activity of the longer chain cations. We have also demonstrated the first use of electrocapillary measurements to study the polarizable RTIL/aqueous interfaces. From the electrocapillary data, the potential of zero charge (PZC) for these RTIL/aqueous interfaces was determined, as well as the relative surface excess charge and capacitance. The PZC was found to be dependent upon the structure of the anions and cations with PZC values ranging from -357 mV for C6mimBETI and -161 mV for C10mimBMSI. The electrocapillary results also show that the cations of the RTIL are becoming increasingly surface-active as the alkyl chain on the cation is lengthened, thereby modulating the interfacial potential. PMID- 16342991 TI - Inhibitive properties and surface morphology of a group of heterocyclic diazoles as inhibitors for acidic iron corrosion. AB - The heterocyclic diazoles 3-amino-1H-isoindole, indazole, imidazole, 4 bromoimidazole, 4-methylimidazole, pyrazole, 4-nitropyrazole, and 4-sulfopyrazole were investigated as corrosion inhibitors of iron in 1 M HCl using ac and dc techniques. The polarization curves showed a decrease in corrosion current for the inhibitor-containing solution. Impedance spectra demonstrate that the charge transfer resistance in the presence of these inhibitors was greater than in inhibitor-free solution, except for 4-nitropyrazole. The resistance increased with inhibitor concentration and with immersion time. The structural and electronic parameters of these diazoles were calculated using computational methodologies. The elemental composition and the speciation of the treated surfaces were investigated via XPS measurements, and morphological changes were monitored by vertical scanning interferometery. PMID- 16342992 TI - Odd and even model self-assembled monolayers: links between friction and structure. AB - The friction between an amorphous carbon tip and two n-alkane monolayers has been examined using classical molecular dynamics simulations. The two monolayers have the same packing density, but the chains comprising each monolayer differ in length by one -CH2- unit. The simulations show that the monolayers composed of C13 chains have higher friction than those composed of C14 chains when sliding in the direction of chain cant; the difference in friction becomes more pronounced as the load is increased. Examination of the contact forces between the chains and the tip, along with conformational differences between the two chain types, lends insight into the friction differences. PMID- 16342993 TI - Effects of hydraulic pressure on the stability and transition of wetting modes of superhydrophobic surfaces. AB - The underlying mechanisms of stability, metastability, or instability of the Cassie-Baxter and Wenzel wetting modes and their transitions on superhydrophobic surfaces decorated with periodic micropillars are quantitatively studied in this article. Hydraulic pressure, which may be generated by the water-air interfacial tension of water droplets or external factors such as raining impact, is shown to be a key to understanding these mechanisms. A detailed transition process driven by increasing hydraulic pressure is numerically simulated. The maximum sustainable or critical pressure of the Cassie-Baxter wetting state on a pillarlike microstructural surface is formulated for the first time in a simple, unified, and precise form. This analytic result reveals the fact that reducing the microstructural scales (e.g., the pillars' diameters and spacing) is probably the most efficient measure needed to enlarge the critical pressure significantly. We also introduce a dimensionless parameter, the pillar slenderness ratio, to characterize the stability of either the Cassie-Baxter or the Wenzel wetting state and show that the energy barrier for transitioning from the Cassie-Baxter to the Wenzel wetting mode is proportional to both the slenderness ratio and the area fraction. Thus, the Cassie-Baxter wetting mode may collapse under a hydraulic pressure lower than the critical one if the slenderness ratio is improperly small. This quantitative study explains fairly well some experimental observations of contact angles that can be modeled by neither Wenzel nor Cassie Baxter contact angles and eventually leads to our proposals for a mixed (or coexisting) wetting mode. PMID- 16342994 TI - Superhydrophobic CFx coating via in-line atmospheric RF plasma of He-CF4-H2. AB - Stable superhydrophobic coatings on various substrates are attained with an in line atmospheric rf plasma process using CF4, H2, and He. The coating layer is composed of CFx nanoparticulates and has an average roughness of approximately 10 nm. This roughness is much smaller than other surfaces reported for superhydrophobicity in the literature. The superhydrophobic coatings are produced on both metallic and insulating substrates without any need of separate microroughening or vacuum lines. PMID- 16342995 TI - Electroactuation of fluid using topographical wetting transitions. AB - The complex morphologies of liquids on topographically structured substrates are exploited for liquid actuation in open microchannels. The liquid is either confined in prefabricated grooves, thus forming elongated filaments, or gathers in macroscopic drops without invading the grooves, depending on conditions. Using the electrowetting effect, we can reversibly switch between these two states. The length of the filaments is sensitive to the ionic content of the liquid and can be described quantitatively with an electrical model considering the voltage drop along the groove. PMID- 16342996 TI - Effect of headgroup size on permeability of Newton black films. AB - Gas permeability and thin-film interferometry are used as a tool to elucidate the orientation of polymeric headgroups in free-standing foam films. Nonionic polyoxyethylene (EO) surfactants were used to stabilize the foam films, keeping the size of the hydrophobic part constant (C12) and varying the size of the hydrophilic (EO numbers) part. The effect of headgroup size on the gas permeability of Newton black foam films was studied. Thickness, contact angle, and surface tension were measured to understand the permeation mechanism. Increase of film thickness and surface tension was observed while increasing the headgroup size, but the contact angle remains small and constant. Upon increasing the headgroup size, the permeability decreases showing that the headgroups provide a resistance to permeation. For smaller headgroups, the permeability follows a linear dependence on the film thickness, whereas for larger headgroups, the permeability essentially deviates from linearity. We use the conventional "coil model" of the EO chains to explain the observed results providing a detailed picture of the orientation of this important molecule in a confined volume of foam films. PMID- 16342997 TI - High catalytic activity of platinum nanoparticles immobilized on spherical polyelectrolyte brushes. AB - We present a study on the catalytic activity of platinum nanoparticles immobilized on spherical polyelectrolyte brushes that act as carriers. The spherical polyelectrolyte brushes consist of a solid core of poly(styrene) onto which long chains of poly(2-methylpropenoyloxyethyl) trimethylammonium chloride are grafted. These positively charged chains form a dense layer of polyelectrolytes on the surface of the core particles ("spherical polyelectrolyte brush") that tightly binds divalent PtCl6-(2) ions. The reduction of these ions within the brush layer leads to nearly monodisperse nanoparticles of metallic platinum. The average size of the particles is approximately 2 nm. The composite particles exhibit excellent colloidal stability. The catalytic activity is investigated by photometrically monitoring the reduction of p-nitrophenol by an excess of NaBH4 in the presence of the nanoparticles. The kinetic data could be explained by the assumption of a pseudo-first-order reaction with regard to p nitrophenol. In all cases, a delay time t0 has been observed, after which the reactions start. This time is shorter when the catalyst has already been used. All data demonstrate that spherical polyelectrolyte brushes present an ideal carrier system for metallic nanoparticles. PMID- 16342998 TI - Wetting of structured hydrophobic surfaces by water droplets. AB - Super-hydrophobic surfaces may arise due to an interplay between the intrinsic, relatively high, contact angle of the more or less hydrophobic solid surface employed and the geometric features of the solid surface. In the present work, this relationship was investigated for a range of different surface geometries, making use of surface free energy minimization. As a rule, the free energy minima (and maxima) occur when the Laplace and Young conditions are simultaneously fulfilled. Special effort has been devoted to investigating the free energy barriers present between the Cassie-Baxter (heterogeneous wetting) and Wenzel (homogeneous wetting) modes. The predictions made on the basis of the model calculations compare favorably with experimental results presented in the literature. PMID- 16342999 TI - Small-angle neutron scattering studies of an adsorbed non-ionic surfactant (C12E24) on hydrophobised silica particles in water. AB - Small-angle neutron scattering (SANS) studies of aqueous dispersions of Stober silica particles (which have been hydrophobised by having 1-octadecanol grafted to their surface), carrying an adsorbed layer of the nonionic surfactant C12E24, in water, have been performed as a function of temperature. Using mixtures of D2O and H2O, the composition of the continuous phase was adjusted to have the same scattering length density as the silica particles. Hence, only the scattering from the 1-octadecanol and C12E24 layers was detected. The data have been analyzed using both a surface Guinier analysis and a two-layer structure model. It has been found that a step profile best describes the inner combined adsorbed layer (1-octadecanol grafted chains, plus the penetrating alkyl chains from the surfactant) and a semi-Gaussian profile the extended poly(ethylene oxide) outer layer. Both analyses demonstrated that the combined surface layer contracted with increasing temperature. PMID- 16343000 TI - Adsorption of non-ionic surfactants on hydrophobic silica particles and the stability of the corresponding aqueous dispersions. AB - The temperature stability of aqueous dispersions of hydrophobic monodisperse silica particles stabilized with nonionic surfactants has been investigated. Adsorption isotherms in conjunction with surface tension measurements showed that the surfactant formed a monolayer on the surface of the particles, where the adsorbed amount depended on the molecular weight of the ethylene oxide headgroup. The temperature stability of these dispersions has been measured by a standard turbidimetric technique and visual observations in terms of their critical flocculation temperature (CFT). Parameters controlling the CFT of the individual dispersions stabilized with a monolayer of surfactant include the thickness of the steric layer, the particle size, and the volume fraction of the particles. Calculations show that the van der Waals attraction between the particles with adsorbed polymer layers increases as the temperature of the dispersion increases, and this largely accounts for the observed CFT behavior. PMID- 16343001 TI - Effect of the carbon surface layer chemistry on benzene adsorption from the vapor phase and from dilute aqueous solutions. AB - We present a complex study of benzene adsorption on chemically modified commercial activated carbons. The porosity of studied carbons is almost the same, whereas the chemical composition and the acid-base properties of surface layers differ drastically from amphoteric (initial de-ashed carbon D43/1, Carbo-Tech, Essen, Germany) and acidic (carbon modified with concentrated HNO3 and fuming H2SO4) to strongly basic (carbon modified with gaseous NH3). Benzene adsorption isotherms measured from aqueous solution at three temperatures (298, 313, and 323 K) and at the neutral pH level are reported. They are supported by studies of water and benzene adsorption from the gaseous phase (volumetric and calorimetric data) and the data of benzene temperature-programmed desorption (TPD). Moreover, the data of the enthalpy of immersion in water and benzene are also presented. Obtained data of benzene adsorption from the gaseous phase are approximated by applying the method of Nguyen and Do (ND) and the Dubinin-Astakhov (DA) equation. The data of adsorption from solution are described by the hybrid DA-Freundlich (DA-F) model. We show that there are similarities in the mechanisms of benzene adsorption from the gaseous phase and from aqueous solutions and that the pore blocking effect is the main stage of the adsorption mechanism. This effect strongly depends on the polarity of the carbon surface. The larger the ratio of the enthalpy of carbon immersion in water to the enthalpy of immersion in benzene, the larger the reduction in adsorption from solution, compared to that in the gaseous phase, that is observed. PMID- 16343002 TI - Effects of metal coating on self-assembled monolayers on gold. 2. Copper on an oligo(phenylene-ethynylene) monolayer. AB - We report studies on the modifications induced by the evaporation of copper overlayers on a self-assembled monolayer (SAM) of the oligo(phenylene-ethynyl) dithiol, 1-thio-4-[4'-[(4'-thio)phenylethynyl]-1'-ethynyl]-benzene (TTPEB). These SAMs were characterized after deposition from a tetrahydrofuran solution on polycrystalline gold substrates and after copper evaporation and its subsequent removal by nitric acid. Monolayers were studied via cyclic voltammetry (CV), UV vis multiwavelength ellipsometry, external reflectance infrared (IR) spectroscopy, and ion scattering spectroscopy (ISS). The results obtained indicate that TTPEB SAMs display the same packing characteristics before and after copper evaporation and removal. However, as shown by IR spectroscopy, the monolayers undergo a reorganization process that involves an increase in tilt angle accompanied by rotation of aromatic rings that results in a decrease in the average molecular twist angle. ISS data suggest that copper diffuses through the monolayer after copper evaporation, a result that is significant for applications of this molecule in molecular electronic devices. PMID- 16343003 TI - Azobenzene-containing monolayer with photoswitchable wettability. AB - A compact monolayer containing azobenzene has been prepared on silicon substrates. The elaboration route consisted of covalent grafting of freshly synthesized azobenzene moieties onto an isocyanate-functionalized self-assembled monolayer (SAM). The highly packed and ordered isocyanate-functionalized SAM and the azobenzene-functionalized SAM were monitored and characterized by contact angle measurements and X-ray reflectivity (XR). Photoswitching of the wettability of the film induced by the reversible cis-trans isomerization of the azobenzene chromophores is experimentally shown from water and olive oil contact angle measurements. PMID- 16343004 TI - Role of exchangeable cations on geometrical and energetic surface heterogeneity of kaolinites. AB - Textural and energetic proprieties of kaolinite were studied by low-pressure argon adsorption at 77 K. The heterogeneity of four kaolinites (two low-defect and two high-defect samples) modified on their surface by cation exchange with Li+, Na+, or K+ was studied by DIS analysis of the derivative argon adsorption isotherms. The comparison between the derivative adsorption isotherms shows that the nature of the surface cation influences the adsorption phenomena on edge and basal faces. In the case of basal faces, two adsorption domains are observed: for the first one, argon adsorption is slightly sensitive to the nature of the surface cation; for the second one, argon adsorption energy depends on the nature of surface cation suggesting their presence on theoretically uncharged basal faces. This study also shows that the shape of elementary particles, as derived from basal and edge surface areas, changes with the nature of cation. This anomalous result is due to the decrease of edge surface area with increasing the size of the cation. This surface cation dependence can be accounted for the area occupied by the edge surface cations in the first argon monolayer. PMID- 16343005 TI - Electric-field-induced patterns in thin polymer films: weakly nonlinear and fully nonlinear evolution. AB - A thin polymer melt on a substrate can be unstable to an electric field normal to the interface, a phenomenon that can be harnessed as a patterning technique with a range of potential applications. Motivated by the variety of patterns observed in experiments for polymers under both unpatterned and patterned masks, we describe here, from theoretical and numerical analyses, how nonlinear effects govern the growth of the instability and determine the final patterns. In particular, we discuss the nonlinear growth in terms of interactions among different Fourier modes and show that the second- and third-order nonlinearities favor the growth of hexagonal patterns under a featureless mask, in agreement with experimental observations. Also, numerical simulations based on the fully nonlinear model validate the prediction of the weakly nonlinear analysis: hexagonal patterns do emerge under an unpatterned mask. Furthermore, in one dimensional simulations, we demonstrate the energetic evolution of this patterning process and reveal several "kinetically stable structures" along the path to the thermodynamically stable state. Two-dimensional simulations allow us to study the effects of both mask patterns and the initial film thickness. Generally, patterns on the mask guide the growth such that the pattern conforms to the geometric shapes. Interestingly, a small cylindrical protrusion at the center of the mask can produce exactly the same pattern as a large, flat, circular protrusion. The initial film thickness or the thickness ratio of the polymer layer to the air gap plays an important role in determining the final pattern formed. Finally, we demonstrate, by two simple examples, that the simulations can provide insights on "smart" mask designs for producing large areas of well-ordered patterns. PMID- 16343006 TI - Electrochemical synthesis of a polypyrrole thin film with supercritical carbon dioxide as a solvent. AB - A conductive polypyrrole (PPy) film was successfully synthesized in a homogeneous supercritical carbon dioxide (scCO2)/acetonitrile (AN) system. The occurrence of a homogeneous supercritical state was confirmed by observations of the phase behavior of the system through a high-pressure cell with a viewing window. The concentration of a supporting electrolyte, tetrabutylammonium hexafluorophosphate (TBAPF6), significantly changed the phase behavior of the scCO2/AN system. The polymerization rate of the film in that system decreased with further addition of CO2. This result suggested that the low viscosity of scCO2 did not play an important role in improving the growth rate of the PPy film. The low polymerization rate might have been due to the electron-transfer resistance arising from the low dielectric constant of scCO2/AN mixture. The roughness of the film prepared in the homogeneous scCO2/AN system was 1/10 that synthesized in AN itself as a solvent. The slow growth of film and the high diffusion rate of the monomer seemed to account for the smooth flat film formation. PMID- 16343007 TI - Tuning cell adhesion on gradient poly(2-hydroxyethyl methacrylate)-grafted surfaces. AB - A simple yet versatile method was developed to prepare a low-density polymerization initiator gradient, which was combined with surface-initiated atom transfer radical polymerization (ATRP) to produce a well-defined poly(2 hydroxyethyl methacrylate) (HEMA) gradient substrate. A smooth variation in film thickness was measured across the gradient, ranging from 20 A to over 80 A, but we observed a nonmonotonic variation in water contact angle. Fits of X-ray reflectivity profiles suggested that at the low graft density end, the polymer chain structure was in a "mushroom" regime, while the polymer chains at high graft density were in a "brush" regime. It was found that the "mushroom" region of the gradient could be made adhesive to cells by adsorbing adhesion proteins, and cell adhesion could be tuned by controlling the density of the polymer grafts. Fibroblasts were seeded on gradients precoated with fibronectin to test cellular responses to this novel substrate, but it was found that cell adhesion did not follow the expected trend; instead, saturated cell adhesion and spreading was found at the low grafting density region. PMID- 16343008 TI - Role of nonadsorbing polymers in bacterial aggregation. AB - Bacteria exhibit properties similar to those of nonbiological colloids and can display pairwise attractions when in close proximity. This interaction is governed by the surface chemistry of the cells. We seek to understand bacterial aggregation at the cellular level using Escherichia coli (E. coli) AB1157. Aggregation studies were carried out using 0.5 to 2.5 wt% E. coli AB1157 harvested in different growth phases with varying concentrations of a nonadsorbing polymer, sodium polystyrene sulfonate (SPS). The electrophoretic mobility of E. coli AB1157 in different growth phases was determined using phase amplitude light scattering. E. coli AB1157 was found to be negatively charged, and the cell surface properties changed in different growth phases. The electrokinetic results correlated well with the different concentrations of nonadsorbing polymer needed to induce depletion aggregation. This shows that a difference in aggregation properties is due to changes in the bacteria electrokinetic properties during their growth. PMID- 16343009 TI - Reducing nonspecific adhesion on cross-linked hydrogel platforms for real-time immunoassay in serum. AB - Biointerfaces that limit nonspecific adhesion of serum proteins have been developed by relying solely on cross-linked hydrogels. In addition to being characterized for adhesion of serum proteins, immunoassay sensitivity was also investigated through a sandwich assay for rhIL-1ra. Among the compositions developed, the optimal surface is comprised of pre-cross-linked carboxymethylcellulose (CMC) and polyethyleneimine (PEI) overlaid on a cross linked layer of poly(ethylene glycol) (PEG) and PEI and employs an anti-IgG Fc specific ligand for oriented antibody immobilization; viscoelastic modeling provides a thickness estimate of 5 nm for the hydrogel alone, rising to 33 nm after the deposition of antibodies. Alternate compositions employing a Protein A ligand and PEG at the exposed surface of the biointerface were disfavored due to an 8-fold increase in serum adhesion and retarded immobilization kinetics, respectively. Through the rapid deposition provided by hydrogels, construction of the entire biointerface, including receptor immobilization, can be completed in 1 h. Based on QCM-D measurements, estimated nonspecific serum adsorption using these compositions is as low as 1.1 ng/mm2. The immunoassay as developed requires 10 min, providing a detection limit of 500 ng/mL rhIL-1ra in 25% human serum using only 5 microg of the secondary antibody. PMID- 16343010 TI - Influence of PEG architecture on protein adsorption and conformation. AB - Poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG) copolymers with various grafting ratios were adsorbed to niobium pentoxide-coated silicon wafers and characterized before and after protein adsorption using X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Three proteins of different sizes, myoglobin (16 kD), albumin (67 kD), and fibrinogen (340 kD), were studied. XPS was used to quantify the amount of protein adsorbed to the bare and PEGylated surfaces. ToF-SIMS and principal component analysis (PCA) were used to study protein conformational changes on these surfaces. The smallest protein, myoglobin, generally adsorbed in higher numbers than the much larger fibrinogen. Protein adsorption was lowest on the surfaces with the highest PEG chain surface density and increased as the PEG layer density decreased. The highest adsorption was found on lysine-coated and bare niobium surfaces. ToF-SIMS and PCA data evaluation provided further information on the degree of protein denaturation, which, for a particular protein, were found to decrease with increasing PEG surface density and increase with decreasing protein size. PMID- 16343011 TI - Tailoring the porosity and morphology of gelatin-methacrylate polyHIPE scaffolds for tissue engineering applications. AB - Gelatin is a natural protein with many desirable properties for application as a biomaterial, including scaffolding for tissue engineering. In this work gelatin A with a molecular weight in the range 50-100 kg mol-1 was modified with methacrylic anhydride and processed into a concentrated oil-in-water emulsion. Polymerization of the continuous phase gave rise to a polyHIPE, a porous material possessing a highly interconnected, trabecular morphology. In the paper, we focused on the goal of obtaining matrixes characterized by suitable sizes of both voids and interconnects, to allow an in depth colonization from transplanted cells. In this respect, we investigated the role of the volume percentage of the dispersed phase and the effect of additives. It was established that high pore volumes (>or=90%) are to be preferred, because they allow the production of solid foams characterized by average void and interconnect diameters of approximately 20 and 10 microm, respectively. These values are still inadequate for the intended application of these scaffolds but represent a good starting point for further improvements. These were achieved through the use of additives, namely sodium chloride and dimethyl sulfoxide, which partially destabilized the precursor emulsion and allowed a solid foam to be obtained with void and interconnect diameters in the range of 30-150 microm and 10-50 microm, respectively. PMID- 16343012 TI - Colloid probe AFM investigation of the influence of cross-linking on the interaction behavior and nano-rheology of colloidal droplets. AB - The repulsive forces between a glass sphere and immobilized colloidal droplets of poly(dimethylsiloxane) (PDMS) (with various levels of internal cross-linking) have been determined in aqueous solution using colloid probe atomic force microscopy. On initial surface approach, droplet deformation is negligible and interaction forces resemble those expected for electrical double layer interaction of rigid spheres. Upon further approach, droplet flattening results in forces that deviate below rigid body electrical double layer interaction. The extent of droplet deformation has been determined in terms of the deviation from hard-sphere interaction. Droplet deformability is strongly dependent on the droplet cross-linking level and hence controlled by some combination of the bulk rheological and interfacial properties of the droplets. Droplet nano-rheology has been determined from the extent of force curve hysteresis. For liquidlike droplets, with low levels of cross-linking, no force curve hysteresis is observed and the elastic deformation may be described by a single spring constant, which is controlled by the interfacial properties. For highly cross-linked droplets, the extent of deformation is controlled by the droplet's bulk rheology rather than the interfacial properties. Upon retraction of the surfaces, force curve hysteresis is observed and is due to the viscoelastic response of the PDMS. The extent of hysteresis is dependent on the rate of approach/retraction and the loading force and has been theoretically analyzed to determine nano-rheological parameters that describe droplet relaxation processes. Elastic moduli and relaxation times of the PDMS droplets vary over several orders of magnitude as a function of cross-linking. PMID- 16343013 TI - Investigation of the core-shell interface in gold@silica nanoparticles: a silica imprinting approach. AB - The nature of the self-assembled core-shell interface in gold@silica nanoparticles synthesized via a 3-aminopropyltrimethoxysilane (APTMS) route is investigated using materials synthesis as a sensitive tool for elucidating interfacial composition and organization. Our approach involves condensation of the gold@silica nanoparticles within a silica framework for synthesis of a composite gold-silica material containing approximately 30 wt % gold. This material contains one of the highest gold loadings reported, but maintains gold core isolation as ascertained via a single surface plasmon resonance absorption band frequency corresponding to that of gold nanoparticles in dilute aqueous solution. The immobilized gold cores are subsequently etched using cyanide anion for the synthesis of templated porosity, which corresponds to the space that was occupied by the gold. Characterization of immobilized amines is performed using probe molecule binding experiments, which demonstrate a lack of accessible amines after gold removal. Solid-state 13C CPMAS NMR spectroscopy on these materials demonstrates that the amount of amine immobilization must be less than 10% of the expected yield, assuming that all of the APTMS becomes bound to the gold nanoparticle template. These results require a core-shell interface in the gold@silica nanoparticles that is predominantly occupied by inorganic silicate species, such as Si-O-Si and Si-OH, rather than primary amines. Such a result is likely a consequence of the weak interaction between primary amines and gold in aqueous solution. Our method for investigating the core-shell interface of gold@silica nanoparticles is generalizable for other interfacial structures and enables the synthesis of bulk imprinted silica using colloidal templates. PMID- 16343014 TI - Oxidative energy storage ability of a TiO2-Ni(OH)2 bilayer photocatalyst. AB - A TiO2-coated indium tin oxide electrode was further coated with Ni(OH)2 by electrodeposition to obtain a TiO2-Ni(OH)2 bilayer film. Upon irradiation of the bilayer film with UV light in a pH 10 buffer, the Ni(OH)2 layer was oxidized, and it turned from colorless to brown; oxidative energy was stored in the layer. The potential of the oxidative energy thus stored was about +0.7 V versus Ag|AgCl. The stored energy could be reversibly taken out of the film by chemical and electrochemical means. The photooxidized Ni(OH)2 was rereduced by alcohols, aldehydes, phenol, I-, H2O2, formate, and acetone. PMID- 16343015 TI - Porosity and mechanical properties of mesoporous thin films assessed by environmental ellipsometric porosimetry. AB - Mesoordered silica thin films with cubic structures were prepared by evaporation induced self-assembly (EISA) with two types of structuring agent (CTAB and block copolymer F127). A complete and accurate description of these films was obtained by combining 2D-SAXS analyses, variable angle spectroscopic ellipsometry, and a specially designed environmental ellipsometric porosimetry (EEP) experiment. The EEP analysis is rapid and cheap and operates at ambient pressure and temperature. This latter experiment was performed with water and produced a set of water adsorption-desorption isotherms. A modified Kelvin equation, coupled with a modelisation of pores contraction, enabled the determination of the structural parameters of films porous networks: ellipsoidal pore diameters, porous volume, and surface area. Young moduli of films in the direction perpendicular to the substrates were calculated from these parameters. PMID- 16343016 TI - Polyelectrolyte multilayers and degradable polymer layers as multicompartment films. AB - Polyelectrolyte multilayers are now a well established concept with numerous potential applications in particular as biomaterial coatings. To timely control the biological activity of cells in contact with a substrate, multicompartment films made of different polyelectrolyte multilayers deposited sequentially on the solid substrate constitute a promising new approach. In a first paper (Langmuir 2004, 20, 7298) we showed that such multicompartment films can be designed by alternating exponentially growing polyelectrolyte multilayers acting as reservoirs and linearly growing ones acting as barriers. In the present study, we first demonstrate however that these barriers composed of synthetic polyelectrolytes are not degraded despite the presence of phagocytic cells. We propose an alternative approach where exponentially growing poly(L lysine)/hyaluronic acid (PLL/HA) multilayers, used as reservoirs, are alternated with biodegradable polymer layers consisting in poly(lactic-co-glycolic acid) (PLGA) and acting as barriers for PLL chains that diffuse within the PLL/HA reservoirs. We first show that these PLGA layers can be deposited alternatively with PLL/HA multilayers leading to polyelectrolyte multilayer/hydrolyzable polymeric layer films and acting as a reservoirs/barriers system. Bone marrow cells seeded on these films ending by a PLL/HA reservoir rapidly degrade it and internalize the PLL chains confined in this reservoir. Then the cells degraded locally the PLGA barrier and internalize the PLL localized in a lower (PLL/HA) compartment after 5 days of seeding. By changing the thickness of the PLGA layer, we hope to be able to tune the time delay of degradation. Such mixed architectures made of polyelectrolyte multilayers and hydrolyzable polymeric layers could act as coatings allowing us to induce a time scheduled cascade of biological activities. We are currently working on the use of comparable films with compartments filled by proteins or peptides and in which the degradation of the barriers results from a hydrolysis over tunable time scales. PMID- 16343017 TI - Assessment of the 3D localization of metallic nanoparticles in Pd/SiO2 cogelled catalysts by electron tomography. AB - The purpose of this study is to analyze the localization of palladium nanoparticles within their silica support, in two heterogeneous catalysts synthesized by the sol-gel process, with different metal loadings. Electron tomography demonstrates that the palladium particles are localized deep inside the silica skeleton. The use of digital image analysis further shows that the dispersion of palladium is optimal in the sample with the lowest loading. The particles are regularly spaced in the middle of the silica skeleton, with a distance between them comparable to the diameter of the struts of silica. PMID- 16343018 TI - Amphiphilic templating of magnesium hydroxide. AB - The synthesis of lamellar mesostructured Mg(OH)2 was achieved through a surfactant templating route. Amphiphilic compounds with different anionic headgroups (phosphate, sulfate, sulfonate, and carboxylate) were used as surfactants. Control of d spacing was achieved through the use of different alkyl carboxylate amphiphiles. It is proposed that the interaction between the highly reactive oxygen atoms of the anionic surfactants and the highly electrophilic Mg atom leads to the formation of high charge density at the interface between the surfactant molecules and the inorganic precursor. This interaction is very strong and the existence of strong bonds between the headgroup molecules of the surfactant and the Mg atom locks the structure in a preferred orientation, i.e., lamellar mesostructure. The strong interaction thus precludes any phase transformation, and only the lamellar phase of Mg(OH)2 is obtained. Calcination of the surfactant by heating in oxygen flow leads to the collapse of the lamellar mesophase and results in the formation of nonporous MgO. PMID- 16343019 TI - Microporous nanocomposites of Pd and Au nanoparticles via hierarchical self assembly. AB - Microporous nanocomposites of Pd and Au nanoparticles were generated by utilizing electrostatic interaction between oppositely charged Au nanoparticles coated with carboxylate groups (Au-COO-) and spherical aggregates of Pd nanoparticles (Pd- NH3+) with a mean diameter of 80+/-20 nm stabilized and cross linked by octa(aminopropyl)silsesquioxane octahydrochloride (POSS-NH3+). Amide bonds were formed between the reactive ion couples that are well defined in the Pd-Au colloidal nanocomposites during a subsequent chemical reaction to generate more stable nanocomposites with improved chemical and physical properties. PMID- 16343020 TI - Simple, one-step synthesis of gold nanowires in aqueous solution. AB - A simple procedure to synthesize gold nanowires based on the reduction of hydrogen tetrachloroaurate by 2-mercaptosuccinic acid in aqueous solution is presented. This procedure requires no additional capping or reduction agent and produces wires with an apparent curly morphology several micrometers in length with diameters as thin as 15 nm. Some of the wires produced end in a ribbonlike structure, finally terminated by a flat triangular prism. Investigations by scanning electron microscopy, transmission electron microscopy (bright and dark field), scanning transmission electron microscopy, and atomic force microscopy as well as conductivity measurements indicate fully connected, polycrystalline gold objects. PMID- 16343021 TI - Density functional theory study of the geometry, energetics, and reconstruction process of Si111 surfaces. AB - We report the structures and energies from first principles density functional calculations of 12 different reconstructed (111) surfaces of silicon, including the 3x3 to 9x9 dimer-adatom-stacking fault (DAS) structures. These calculations used the Perdew-Burke-Ernzerhof generalized gradient approximation of density functional theory and Gaussian basis functions. We considered fully periodic slabs of various thicknesses. We find that the most stable surface is the DAS 7x7 structure, with a surface energy of 1.044 eV/1x1 cell (1310 dyn/cm). To analyze the origins of the stability of these systems and to predict energetics for more complex, less-ordered systems, we develop a model in which the surface energy is partitioned into contributions from seven different types of atom environments. This analysis is used to predict the surface energy of larger DAS structures (including their asymptotic behavior for very large unit cells) and to study the energetics of the sequential size change (SSC) model proposed by Shimada and Tochihara for the observed dynamical reconstruction of the Si(111) 1x1 structure. We obtain an energy barrier at the 2x2 cell size and confirm that the 7x7 regular stage of the SSC model (corresponding to the DAS 7x7 reconstruction) provides the highest energy reduction per unit cell with respect to the unreconstructed Si111 1x1 surface. PMID- 16343023 TI - Oral contraceptives and skin cancer: is there a link? AB - The skin expresses estrogen, progesterone, and androgen receptors. In the presence of steroid hormones, such as those contained in oral contraceptives, the skin likely responds to hormonal signals that control the cell cycle, apoptosis, DNA replication, and other cellular functions. Some estrogen-responsive pathways have the potential to promote tumor development, including the augmentation of epidermal growth factor signaling, the expression of proto-oncogenes, and inhibition of apoptosis. The question of whether oral contraceptives increase the risk for the development of skin cancer, particularly melanoma, is still an area of concern. This paper reviews the available evidence, the bulk of which suggests that while the skin responds to estrogens, progestins, and androgens, these responses do not significantly increase the risk of developing skin cancer when estrogen exposure is not excessive. PMID- 16343024 TI - Monitoring liver function during methotrexate therapy for psoriasis: are routine biopsies really necessary? AB - Methotrexate is a widely used and efficacious treatment for psoriasis. Current recommendations advise liver biopsy after each cumulative 1-1.5g dosage of methotrexate. Although studies of methotrexate hepatotoxicity in psoriasis are of variable quality, the evidence indicates that in patients with no risk factors for liver disease on long-term, low-dosage (<20mg), once-weekly methotrexate, liver biopsy is justified after an initial cumulative dosage of 4g. Advances in the non-invasive assessment of liver fibrogenesis may eventually further reduce the need for biopsy. PMID- 16343025 TI - Human papillomavirus: burden of illness and treatment cost considerations. AB - The monetary and personal costs to society of human papillomavirus (HPV) infection are enormous. In order to make a comparison of different treatment methods we reviewed the entire literature on HPV treatment from January 1966 to December 2003 using MEDLINE, with particular reference to published meta analyses, randomized controlled and comparative studies. Patient-applied therapies offer patients the possibility of convenient and, on the whole, pain free treatment. Podofilox (podophyllotoxin) and salicylic acid for genital and extragenital warts, respectively, have the additional advantage of being the most cost-effective treatments and, on this basis, they are to be commended as appropriate first-line agents. The second-line treatment of choice for common warts is cryotherapy. For recalcitrant common warts possible options include inosine pranobex with cryotherapy or electrosurgery, imiquimod with paring and occlusion, intralesional bleomycin, or diphencyprone. Alternative first-line and second-line treatments for genital warts would be either some form of surgical removal or imiquimod. The first option may be the cheapest but this has to be balanced against a degree of post-operative morbidity. Limited data from comparative studies do not show any clear difference in efficacy between cryotherapy, trichloroacetic acid, scissor excision, electrosurgery, and laser surgery in the treatment of genital warts, and the cost effectiveness of these therapies is probably similar to that of imiquimod. Cryotherapy and trichloroacetic acid are relatively expensive and inconvenient for patients and should be reserved as third-line treatments with certain exceptions, such as cryotherapy for meatal warts. The duration of treatment is significantly related to the number of warts present, the area covered by the warts, and the length of time the warts have been present. For recalcitrant anogenital warts third-line treatment options that show promise include surgery in combination with imiquimod or cidofovir cream. For squamous intraepithelial lesions that cannot easily be excised or physically ablated current treatment options include imiquimod and fluorouracil cream. The latter is an inexpensive option but causes the greatest morbidity. It is hoped that cidofovir may be added to this list if it becomes commercially available, and that protective and therapeutic HPV vaccines will transform the management of HPV in the future. PMID- 16343027 TI - The role of hyaluronic acid in wound healing: assessment of clinical evidence. AB - Hyaluronic acid (hyaluronan), a naturally occurring polymer within the skin, has been extensively studied since its discovery in 1934. It has been used in a wide range of medical fields as diverse as orthopedics and cosmetic surgery, but it is in tissue engineering that it has been primarily advanced for treatment. The breakdown products of this large macromolecule have a range of properties that lend it specifically to this setting and also to the field of wound healing. It is non-antigenic and may be manufactured in a number of forms, ranging from gels to sheets of solid material through to lightly woven meshes. Epidermal engraftment is superior to most of the available biotechnologies and, as such, the material shows great promise in both animal and clinical studies of tissue engineering. Ongoing work centers around the ability of the molecule to enhance angiogenesis and the conversion of chronic wounds into acute wounds. PMID- 16343026 TI - The psychosocial burden of psoriasis. AB - BACKGROUND: Skin diseases such as psoriasis can profoundly influence a patient's self-image, self-esteem, and sense of well-being. Psoriasis is a multifactorial inflammatory condition with a disease burden that extends beyond the physical symptoms experienced by patients. Psoriasis affects all aspects of quality of life, including physical, psychologic, social, sexual, and occupational elements. OBJECTIVE: The goal of this article was to review the published literature on the impact of psoriasis on quality of life. METHODS: Relevant studies were identified through a comprehensive search of MEDLINE, EMBASE, and the Derwent Drug File databases of English-language articles published between 1993 and 2005 using the terms psoriasis in combination with quality of life, cost, cost-benefit analysis, economic, employment, days lost, healthcare, hospitalization, managed care, outcomes research, occupation, payers, and psychosocial. The reference lists of identified articles were checked for additional studies that might have been missed in the original searches. RESULTS: Data suggest that social stigmatization, high stress levels, physical limitations, depression, employment problems and other psychosocial co-morbidities experienced by patients with psoriasis are not always proportional to, or predicted by, other measurements of disease severity such as body surface area involvement or plaque severity. CONCLUSION: It is essential to include measures of psychosocial morbidity when assessing psoriasis severity and treatment efficacy because of the substantial role that psychosocial burden plays in patient perception of disease severity, quality of life, and disease course. PMID- 16343028 TI - Comparison of azelaic acid and anthralin for the therapy of patchy alopecia areata: a pilot study. AB - BACKGROUND: Although topical azelaic acid has been previously used for the treatment of alopecia, no controlled trials of azelaic acid for this condition have been conducted to date. OBJECTIVE: The goal of this study was to determine the efficacy, tolerability, and safety of azelaic acid treatment in patients with patchy alopecia areata (AA) in comparison with anthralin (dithranol) treatment. SUBJECTS AND METHODS: This study included 31 subjects with patchy AA who did not receive any treatment for at least 1 month prior to the study. Demographic and clinical characteristics of these subjects were recorded at baseline. Subjects were randomized to apply either 20% azelaic acid (15 subjects) or 0.5% anthralin (16 subjects) for 12 consecutive weeks. In a subsequent 8-week follow-up period no cream was applied. Two independent investigators performed an efficacy evaluation with clinical examination using a terminal hair regrowth score (RGS) with a scale ranging from 0 (inadequate response) to 2 (complete response) at week 20. Partial response was accepted as score 1. RESULTS: Both groups were well matched for the relevant demographic and clinical indicators affecting treatment response at baseline. All subjects completed the trial. At week 20 the RGS was 1.27 +/- 0.9 in the azelaic acid group versus 1.37 +/- 0.8 in the anthralin group (p > 0.05). A complete response was observed in 53.3% of cases in the azelaic acid group (8 of 15) compared with 56.2% (9 of 16) in the anthralin group (p > 0.05). No serious adverse events were observed in either group during the study. CONCLUSION: The present pilot study showed that the use of azelaic acid gave similar results to anthralin with regard to hair regrowth, and that it can be an effective topical therapy for patchy AA. More extensive trials are necessary, however, to reach a definitive conclusion. PMID- 16343029 TI - Monilethrix: improvement with acitretin. AB - Monilethrix is a rare hereditary disorder that affects the hair and is characterized by shaft anomaly. There is no known treatment that successfully cures the condition. In this report we present a case of monilethrix in a 7-year old girl treated with oral acitretin. A very good clinical and cosmetic result was obtained while treatment was continued. However, clinical symptoms recurred after discontinuation of acitretin therapy. PMID- 16343031 TI - Scaly red plaques on dorsal part of hand. Patient notes morning stiffness and pain. PMID- 16343032 TI - Chronic obstructive pulmonary disease: assessing and treating psychological issues in patients with COPD. PMID- 16343033 TI - Does anemia matter? Anemia, morbidity, and mortality in older adults: need for greater recognition. AB - Anemia is common and under recognized in older adults and associated with increased morbidity and mortality. Estimates of prevalence of anemia in older adults vary considerably based on the setting, gender, age and definition used and likely to increase further based on aging trends. Rather than simply a consequence of aging, anemia is a marker of underlying disease, requiring investigation for an etiology. A cause is discernible in at least two-thirds of cases; management involves addressing the underlying disease process, replacement of deficient nutrients or the use of erythropoietic factors. PMID- 16343034 TI - Epilepsy in older adults. Common morbidities influence development, treatment strategies, and expected outcomes. AB - Given that that the average person has one chronic illness for each decade over age 50, one would expect that patients who develop seizures in late life would have associated medical and/or neurologic conditions. Cerebrovascular disease, hypertension, heart disease, diabetes mellitus, renal disease, and dementia all relate to epilepsy. Co-morbidities not only contribute to the causation and consequences of seizures, they also interfere with effective treatment and optimal functioning. Because seizures in older individuals can lead to serious consequences, safe and effective treatment is essential. Yet, antiepileptic drugs (AEDs) may cause adverse effects that may be worse in older patients when compared to younger patients. Multiple medications lead to a high probability that medically significant drug interactions may occur and must be monitored for in geriatric patients. PMID- 16343035 TI - Differential right and left ventricular diastolic tolerance to acute afterload and NCX gene expression in Wistar rats. AB - This study evaluated right ventricular (RV) and left ventricular (LV) diastolic tolerance to afterload and SERCA2a, phospholamban and sodium-calcium exchanger (NCX) gene expression in Wistar rats. Time constant tau and end diastolic pressure-dimension relation (EDPDR) were analyzed in response to progressive RV or LV afterload elevations, induced by beat-to-beat pulmonary trunk or aortic root constrictions, respectively. Afterload elevations decreased LV- tau, but increased RV-tau. Whereas LV- tau analyzed the major course of pressure fall, RV- tau only assessed the last fourth. Furthermore, RV afterload elevations progressively upward shifted RV EDPDR, whilst LV afterload elevations did not change LV-EDPDR. SERCA2a and phospholamban mRNA were similar in both ventricles. NCX-mRNA was almost 50 % lower in RV than in LV. Left ventricular afterload elevations, therefore, accelerated the pressure fall and did not induce diastolic dysfunction, indicating high LV diastolic tolerance to afterload. On the contrary, RV afterload elevations decelerated the late RV pressure fall and induced diastolic dysfunction, indicating small RV diastolic tolerance to afterload. These results support previous findings relating NCX with late Ca(2+) reuptake, late relaxation and diastolic dysfunction. PMID- 16343036 TI - Hydrogen peroxide metabolism in alveolar macrophages after exposure to hypoxia and heat. AB - High temperature can change the effects of intra- and intercellular regulators and therefore modify the cellular response to hypoxia. We investigated H(2)O(2) production by alveolar macrophages, isolated from adult male rats, which were incubated under conditions of oxygen deficiency and high temperature (experiment in vitro). The incubation of these cells for 2 hours at 10 % or 5 % oxygen led only to slight fluctuations in the H(2)O(2) level, while the rise of temperature from 37 degrees C up to 42 degrees C significantly increased its generation. Level of thiobarbituric acid-reactive substances (TBARS) underwent similar changes. Under these conditions the accumulation of H(2)O(2) was found to be caused mainly by its decreased cleavage rather than its enhanced production. This is indicated by decreased catalase and glutathione peroxidase activity together with a parallel absence of significant changes in superoxide dismutase (SOD) activity. Slight fluctuation of reduced glutathione level and the pronounced increase of glucose-6-phosphate dehydrogenase (G6PD) activity were detected. Strong (5 %) but not moderate (10 %) lack of oxygen led to a sharp increase in formation of cellular nitrite ions by alveolar macrophages. In general, our data showed that high temperature did not lead to any qualitative shifts of defined hypoxia-derived changes in oxidant/antioxidant balance in alveolar macrophages, but promoted sensitivity of cells to oxygen shortage. PMID- 16343037 TI - Protective effect of S-adenosylmethionine against galactosamine-induced injury of rat hepatocytes in primary culture. AB - The protective effect of S-adenosylmethionine (SAMe) on D-galactosamine (GalN) induced damage to rat hepatocytes was tested in primary cultures. SAMe at concentrations of 50 and 1000 mg/l significantly reduced lactate dehydrogenase release from cells injured by 40 mM GalN after 24 h of incubation. There were no significant changes in urea production after 24 h among tested groups, including control hepatocytes. Exposure of hepatocytes to GalN leads to 3.5-fold decrease in urea synthesis after 48 h in comparison with control cell cultures. Addition of the highest dose of SAMe (1000 mg/l) into the culture media attenuated this decrease by 180 %. None of the tested doses of SAMe (5, 25, 50 and 1000 mg/l) affected considerably the reduced activity of mitochondrial dehydrogenases. The content of reduced and oxidized glutathione in GalN-exposed cells was diminished to 1.5 % and 16 %, respectively, of the control values after 24 h. Using only the highest concentration SAMe increased significantly these contents. SAMe had no effect on dramatically decreased albumin synthesis. These findings indicate beneficial effect of SAMe, especially of the highest concentration, on GalN induced toxicity to rat hepatocytes in primary culture. This action of SAMe seems to be associated with reduction of plasma membrane damage and increased synthesis of glutathione. PMID- 16343038 TI - Effect of gene polymorphisms on lipoprotein levels in patients with dyslipidemia of metabolic syndrome. AB - Dyslipidemia in the metabolic syndrome (MS) is considered to be one of the most important risk factors for atherosclerosis. It is characterized by hypertriglyceridemia, low concentration of plasma HDL-cholesterol, predominance of small dense LDL particles and an increased concentration of plasma apolipoprotein B (apoB). The pathogenesis of this type of dyslipidemia is partially explained, but its genetic background is still unknown. To evaluate the influence of cholesterol ester transfer protein (CETP) TaqIB polymorphism, lipoprotein lipase (LPL) PvuII and HindIII polymorphisms, hepatic lipase (LIPC) G 250A polymorphism and apolipoprotein C-III (APOC3) SstI gene polymorphism on lipid levels in dyslipidemia of the metabolic syndrome, 150 patients with dyslipidemia of metabolic syndrome were included. 96 % of patients had type 2 diabetes. The patients did not take any lipid lowering treatment. The exclusion criterion was the presence of any disease that could affect lipid levels, such as thyroid disorder, liver disease, proteinuria or renal failure. Gene polymorphisms were determined using the polymerase chain reaction and restriction fragment length polymorphisms. The genotype subgroups of patients divided according to examined polymorphisms did not differ in plasma lipid levels with the exception of apoB. The apoB level was significantly higher in patients with S1S1 genotype of APOC3 SstI polymorphism when compared with S1S2 group (1.10+/-0.26 vs. 0.98+/ 0.21 g/l, p=0.02). Similarly, patients with H-H- genotype of LPL HindIII polymorphism had significantly higher mean apoB, compared with H+H- and H+H+ group (1.35+/-0.30 vs. 1.10+/-0.26 g/l, p=0.02). In the multiple stepwise linear regression analysis, apoB level seemed to be influenced by APOC3 SstI genotype, which explained 6 % of its variance. The present study has shown that the S1 allele of APOC3 SstI polymorphism and the H- allele of LPL HindIII polymorphism might have a small effect on apoB levels in the Central European Caucasian population with dyslipidemia of metabolic syndrome. PMID- 16343039 TI - Folate co-administration improves the effectiveness of fenofibrate to decrease the lipoprotein oxidation and endothelial dysfunction surrogates. AB - Fibrate therapy results in elevation of plasma total homocysteine (tHcy), which is known to induce oxidative stress and endothelial dysfunction. We aimed to establish whether fibrate-induced elevation of tHcy has also similar consequences and whether they may be prevented by folate co-administration. Eighteen subjects with hypercholesterolemia were included in an open, prospective, cross-over study. We compared intra-individually the effect of fenofibrate on tHcy, oxidative stress and endothelial dysfunction surrogates, in monotherapy and when combined with 10 mg of folate. These effects were also compared with fluvastatin monotherapy. Fenofibrate in monotherapy significantly decreased LDL cholesterol, increased the tHcy by 39.5 %, while oxidized LDL (oxLDL), malondialdehyde (MDA), von Willebrand factors (vWf) and thrombomodulin (TMD) remained unchanged. When fibrate was co-administered with folate, the tHcy remained on the initial post diet level, while both the total and oxLDL as well as MDA, vWf and TMD decreased. In contrast to fenofibrate monotherapy, fluvastatin (80 mg) had a similar effect as combined therapy with fenofibrate and folate, while tHcy remained uninfluenced. In conclusion, fenofibrate decreases the LDL cholesterol, but in contrast to fluvastatin, has no significant antioxidative and endothelium protective potential, probably due to a concomitant increase of tHcy. These effects may be improved by co-administration of folate. PMID- 16343040 TI - Changes in placental adipocytokine gene expression associated with gestational diabetes mellitus. AB - Leptin and adiponectin, two adipocytokines, may work together in regulating energy homeostasis and insulin action. Leptin gene expression has been investigated in term placental tissue complicated by gestational diabetes mellitus (GDM), but never in conjunction with all isoforms of the leptin receptor (LEPR A-D), or with adiponectin receptors (ADIPOR1 and 2). In this study we examined the association between changes in expression of these genes in placental tissue and GDM risk. We assessed placental gene expression of leptin, LEPR A-D and ADIPOR1 and 2 by real time PCR using mRNA from maternal and fetal biopsies. Tissues were collected from uncomplicated pregnancies (n=28) and those complicated by GDM (n=19). Gene expression was normalized to three endogenous housekeeping genes. Relative gene expression values were reported as fold change between groups. Adiponectin gene expression was out of the sensitive range of our assay. There were increases in leptin mRNA expression in GDM cases compared with controls for maternal-side (p=0.06), and fetal-side (p=0.09) placental biopsies. No significant changes were seen in GDM cases compared with controls in LEPR A-D or ADIPOR1 and 2. mRNA derived from maternal-side tissue was positively correlated with tissue from the fetal side for all genes studied (all p<0.01). Finally, we noted that absence or presence of GDM was a major factor in leptin mRNA expression after adjusting for maternal age, mode of delivery, parity and smoking status. In conclusion, increases in leptin mRNA expression in term placenta, but not that of its receptors, are associated with the diagnosis of GDM. Changes seen in the ligand, but not the receptor, of the leptin pathway in GDM-complicated pregnancies may also apply to the adiponectin pathway, as the ADIPOR1 and 2 mRNAs do not change with GDM diagnosis. PMID- 16343041 TI - Early and late allergic phase related cough response in sensitized guinea pigs with experimental allergic rhinitis. AB - Cough is a common and important symptom of asthma and allergic rhinitis. Previous experimental evidence has shown enhanced cough sensitivity during early phase of experimental allergic rhinitis in guinea pigs. We hypothesized that airway inflammation during the late phase response after repeated nasal antigen challenge may affect the afferent sensory nerve endings in the larynx and tracheobronchial tree and may also modulate cough response. In the present study we evaluated the cough sensitivity during a period of early and late allergic response in sensitized guinea pigs after repeated nasal antigen challenges. Forty five guinea pigs were sensitized with ovalbumin (OVA). Four weeks later 0.015 ml of 0.5 % OVA was intranasally instilled to develop a model of allergic rhinitis that was evaluated from the occurrence of typical clinical symptoms. Animals were repeatedly intranasally challenged either by OVA (experimental group) or by saline (controls) in 7-day intervals for nine weeks. Cough was elicited by inhalation of citric acid aerosols. Cough was evaluated at 1 or 3 h after the 6th nasal challenge and 17 or 24 h after the 9th nasal challenge. The cough reflex was significantly increased at 1 and 3 h after repeated nasal challenge in contrast to cough responses evoked at 17 and 24 h after repeated nasal challenge. In conclusion, enhanced cough sensitivity only corresponds to an early allergic response after repeated nasal challenges. PMID- 16343042 TI - Effect of thyroxine on antioxidant defense system in the liver of different aged rats. AB - The effects of altered thyroid state on the antioxidant defense system in the liver of differently aged rats were examined. Male rats aged 15, 45 and 75 days were treated with L-thyroxine, T(4) (40 microg/100 g body mass, s.c., one dose per day) for 14 days (finally aged 30, 60 and 90 days, respectively). The following antioxidant defense enzymes were measured: superoxide dismutases (both copper zinc, CuZn-SOD and manganese containing, Mn-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST), glutathione reductase (GR), as well as the content of low molecular mass antioxidant glutathione (GSH). The effect of T(4) on antioxidant defense system in the liver differs with respect to age. T(4) treatment decreased CAT and GST activities, as well as the content of GSH in animals aged 60 and 90 days. The same treatment elevated GR activity in rats at 30 days of age, this phenomenon was not observed in older animals. The different response of immature rats to thyroxine compared to older animals could be attributed to the differences in thyroxine metabolism and the developmental pattern. Direct effect of T(4) on mature rats can be considered as a part of its overall catabolic action. PMID- 16343043 TI - Hypoxic stress-enhanced expression and release of adrenomedullin (AM) and up regulated AM receptors, while glucose starvation reduced AM expression and release and down-regulated AM receptors in monkey renal cells. AB - The proliferative peptide adrenomedullin (AM) has a wide distribution in a variety of tissues and cells. The mechanism how the AM gene is regulated in cells is not yet known. The renal cortex, renal vascular smooth muscles, glomeruli and tubular epithelial cells are very sensitive to hypoxia. Renal hypoxia produces acute renal tubular necrosis and markedly induces AM expression in damaged cells. However, little information is available regarding the possible pathophysiological production and release of renal tubular AM. Regulation of membrane-bound AM receptors in renal cells has not yet been systematically studied. To elucidate the potential pathological role of human AM we examined the production and release of AM, as well as the characteristics of surface membrane AM receptors in cultured monkey renal tubular epithelial cells (RC) exposed to hypoxia, induced with endothelin-1, and subjected to glucose deprivation. Exposure of RC to hypoxia (1 % O(2), 5 % CO(2) in N(2)), and to phorbol 12 myristate 13-acetate (PMA) increased production and secretion of AM and increased specific [(125)I]AM binding on RC. Metabolic stress (1 % glucose in the cultivation medium) and preincubation of RC with rival peptide endothelin-1 significantly reduced immunoreactive-AM in a conditioned medium and whole cell surface membrane AM binding on RC. Altogether, our data suggest that the AM is involved in the adaptation of renal tubular cells to hypoxia. Increased expression of AM mRNA and regulation of AM receptors in metabolic stress may function as an important autocrine/paracrine regulator(s) of renal tubular epithelial cells. PMID- 16343044 TI - Adenosine potentiates stimulatory effects on granulocyte-macrophage hematopoietic progenitor cells in vitro of IL-3 and SCF, but not those of G-CSF, GM-CSF and IL 11. AB - The aim of the studies was to ascertain if adenosine is able to co-operate with selected hematopoietic growth factors and cytokines, namely with granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), stem cell factor (SCF), interleukin-3 (IL-3), and interleukin-11 (IL-11), in inducing the growth of colonies from hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) from normal bone marrow cells in vitro. Adenosine was found not to produce any colonies when present in the cultures as the only potential stimulator. All the tested cytokines and growth factors were observed to induce the growth of distinct numbers of GM-CFC colonies, with the exception of IL-11. When suboptimal concentrations of the evaluated cytokines and growth factors were tested in the cultures in which various concentrations of adenosine were concomitantly present, mutually potentiating effects were found in the case of IL-3 and SCF. These results confirm the role of adenosine in regulation of granulopoiesis and predict IL-3 and SCF as candidates for further in vivo studies of their combined administration with adenosine. PMID- 16343045 TI - Thermal, cardiac and adrenergic responses to repeated local cooling. AB - The aim of this study was to ascertain whether repeated local cooling induces the same or different adaptational responses as repeated whole body cooling. Repeated cooling of the legs (immersion into 12 degrees C water up to the knees for 30 min, 20 times during 4 weeks = local cold adaptation - LCA) attenuated the initial increase in heart rate and blood pressure currently observed in control subjects immersed in cold water up to the knees. After LCA the initial skin temperature decrease tended to be lower, indicating reduced vasoconstriction. Heart rate and systolic blood pressure appeared to be generally lower during rest and during the time course of cooling in LCA humans, when compared to controls. All these changes seem to indicate attenuation of the sympathetic tone. In contrast, the sustained skin temperature in different areas of the body (finger, palm, forearm, thigh, chest) appeared to be generally lower in LCA subjects than in controls (except for temperatures on the forehead). Plasma levels of catecholamines (measured 20 and 40 min after the onset of cooling) were also not influenced by local cold adaptation. Locally cold adapted subjects, when exposed to whole body cold water immersion test, showed no change in the threshold temperature for induction of cold thermogenesis. This indicates that the hypothermic type of cold adaptation, typically occurring after systemic cold adaptation, does not appear after local cold adaptation of the intensity used. It is concluded that in humans the cold adaptation due to repeated local cooling of legs induces different physiological changes than systemic cold adaptation. PMID- 16343046 TI - Perfusion pressure manipulation in porcine sepsis: effects on intestinal hemodynamics. AB - Limited information is available about selection of the threshold for arterial blood pressure in critically ill patients, particularly in sepsis when normal organ blood flow autoregulation may be altered. The present experimental study investigated whether increasing perfusion pressure using norepinephrine in normotensive hyperdynamic porcine bacteremia affects intestinal macro- and microcirculation. Nine pigs received continuous i.v. administration of Pseudomonas aeruginosa (PSAE) to develop hyperdynamic, normotensive (mean arterial pressure [MAP] 65 mm Hg) sepsis. Norepinephrine was used to achieve 10 15 % increase in MAP. Mesenteric arterial blood flow (Q(gut)), ileal mucosal microvascular perfusion (LDF(gut)) and ileal-end-tidal PCO(2) gap (PCO(2) gap) were measured before norepinephrine, after 60 min of norepinephrine infusion and 60 min after norepinephrine infusion had been discontinued. During a 12 h period of PSAE infusion all pigs developed hyperdynamic circulation with significantly decreased MAP. Although the mesenteric blood flow remained unchanged, infusion of PSAE resulted in a gradual fall of ileal microvascular perfusion, which was associated with progressively rising PCO(2) gap. Norepinephrine which induced a 10-15 % increase in perfusion pressure (i.e. titrated to attain near baseline values of MAP) affected neither Q(gut) nor the intestinal blood flow distribution (Q(gut)/CO). Similarly, norepinephrine did not change either LDF(gut) or PCO(2) gap. In this hyperdynamic, normotensive porcine bacteremia, norepinephrine induced increase in perfusion pressure exhibited neither beneficial nor deleterious effects on intestinal macrocirculatory blood flow and ileal mucosal microcirculation. The lack of changes suggests that the gut perfusion was within its autoregulatory range. PMID- 16343047 TI - Oxidized low-density lipoprotein (Oxidized LDL) and the risk of preeclampsia. AB - Oxidative stress plays an important role in the pathophysiology of preeclampsia. In a case-control study of 99 women with preeclampsia and 99 controls, we assessed maternal plasma oxidized low-density lipoprotein (oxidized LDL) in relation to preeclampsia risk. Logistic regression procedures were used to derive odds ratios (OR) and 95 % confidence intervals (CI). Plasma oxidized LDL was determined using enzyme immunoassay. Maternal plasma oxidized LDL was significantly positively correlated with lipids in both cases and controls. After adjusting for nulliparity, pre-pregnancy body mass index, physical inactivity, family history of chronic hypertension and plasma vitamin C concentrations, women who had elevated oxidized LDL concentrations ( > or = 50 U/l) experienced a 2.9 fold increased risk of preeclampsia when compared with women having lower oxidized LDL concentrations (95 % CI 1.4-5.9). The risk of preeclampsia was markedly increased in women who had both elevated oxidized LDL and elevated triglyceride concentrations (OR=8.9, 95 % CI 3.1-26.2). Women with both elevated oxidized LDL and low vitamin C concentrations experienced a 9.8-fold increased risk of preeclampsia (95 % CI 3.0-32.2). Our results confirm the role of oxidative stress in the pathogenesis of preeclampsia. Prospective studies are needed to determine if elevated oxidized LDL concentrations can predict the occurrence of preeclampsia. PMID- 16343048 TI - Acute pancreatitis: proteinase-activated receptor-2 as Dr. Jekyll and Mr. Hyde. AB - "Proteinase-activated" receptor-2 (PAR-2) is a G protein-coupled transmembrane receptor with seven transmembrane domains activated by trypsin. It has been shown in the pancreatic tissue that PAR-2 is involved in duct/acinary cells secretion, arterial tonus regulation and capillary liquid content turnover under physiological conditions. These above mentioned structures play an important role during the development of acute pancreatitis and are profoundly influenced by a high concentration of trypsin enzyme after its secretion into the interstitial tissue from the basolateral aspect of acinar cells. Among the other factors, it is the increase of interstitial trypsin concentration followed rapidly by PAR-2 action on pancreatic vascular smooth muscle cells that initiates ischemic changes in pancreatic parenchyma and that finally leads to necrosis of the pancreas. Consequent reperfusion perpetuates changes leading to the acute pancreatitis development. On the contrary, PAR-2 action on both exocrine and duct structures seems to play locally a protective role during acute pancreatitis development. Moreover, PAR-2 action is not confined to the pancreas but it contributes to the systemic vascular endothelium and immune cell activation that triggers the systemic inflammatory response syndrome (SIRS) contributing to an early high mortality rate in severe disease. PMID- 16343053 TI - The current and future dynamics of disease in plant communities. AB - Pathogens are powerful evolutionary forces shaping the structure and dynamics of both individual species and of the communities of which they are part, at a broad range of genetic, ecological, spatial, and temporal scales. At all these levels their impact varies from the subtle and little recognized through to the most obvious destruction. Today the direct role of pathogens in natural plant communities is better recognized than at previous times, although the nuances of their interactions and the cascade of ramifications that can flow through changing biotic and abiotic effects are only now gaining recognition. However, as human influence on pathogens increases--either directly through enhanced if accidental dispersal, or through anthropogenic impacts on climate--we may expect to see increasing evidence of pathogens affecting plant species, community structure, and ecosystem function. PMID- 16343054 TI - A quantitative lymphoscintigraphic evaluation of lymphatic function in the swollen hands of women with lymphoedema following breast cancer treatment. AB - In BCRL (breast cancer-related lymphoedema), arm swelling is unevenly distributed and some regions are partly or entirely spared. In particular, the hand may or not be swollen, but when involved functional impairment can be substantial. We have found previously that, when the ipsilateral hand is spared of swelling (in a limb with swelling proximal to the hand), the local lymph drainage rate constant (k) is at least as high as in the contralateral hand, contrary to the traditional 'stopcock' concept of reduced lymph drainage from the whole limb. In the light of this finding, we have investigated lymph drainage in the hands of eight women with BCRL and moderate-to-severe hand swelling, using gamma-camera quantitative lymphoscintigraphy. Images showed pronounced superficial activity in the ipsilateral swollen arms of most patients, indicating dermal backflow. k for 99mTc-labelled hIgG (human IgG) measured over 5 h in the subcutis of the ipsilateral swollen hand was 34+/-24% less than in the contralateral hand (P=0.013). Activity measured in the ipsilateral swollen forearm increased progressively, but there was very little increase in the contralateral forearm, indicating retention of 99mTc-labelled hIgG in the swollen forearm. It is concluded that lymphatic function in the swollen hand is impaired, and that there appears to be two populations of women with BCRL, i.e. spared-hand and swollen hand, irrespective of the cancer treatment received. PMID- 16343055 TI - Peroxisome-proliferator-activated receptor alpha agonists inhibit cyclo-oxygenase 2 and vascular endothelial growth factor transcriptional activation in human colorectal carcinoma cells via inhibition of activator protein-1. AB - Recent evidence indicates that PPAR (peroxisome-proliferator-activated receptor) alpha ligands possess anti-inflammatory and antitumoural properties owing to their inhibitory effects on the expression of genes that are involved in the inflammatory response. However, the precise molecular mechanisms underlying these effects are poorly understood. In the present study, we show that tumour promoter PMA-mediated induction of genes that are significantly associated with inflammation, tumour growth and metastasis, such as COX-2 (cyclo-oxygenase 2) and VEGF (vascular endothelial growth factor), is inhibited by PPARalpha ligands in the human colorectal carcinoma cell line SW620. PPARalpha activators LY-171883 and WY-14,643 were able to diminish transcriptional induction of COX-2 and VEGF by inhibiting AP-1 (activator protein-1)-mediated transcriptional activation induced by PMA or by c-Jun overexpression. The actions of these ligands on AP-1 activation and COX-2 and VEGF transcriptional induction were found to be dependent on PPARalpha expression. Our studies demonstrate the existence of a negative cross-talk between the PPARalpha- and AP-1-dependent signalling pathways in these cells. PPARalpha interfered with at least two steps within the pathway leading to AP-1 activation. First, PPARalpha activation impaired AP-1 binding to a consensus DNA sequence. Secondly, PPARalpha ligands inhibited c-Jun transactivating activity. Taken together, these findings provide new insight into the anti-inflammatory and anti-tumoural properties of PPARalpha activation, through the inhibition of the induction of AP-1-dependent genes that are involved in inflammation and tumour progression. PMID- 16343056 TI - Portal hypertension: from pathophysiology to clinical practice. AB - Portal hypertension (PHT) is responsible for the more severe and often lethal complications of cirrhosis such as bleeding oesophageal varices, ascites, renal dysfunction and hepatic encephalopathy. Because of the combined impact of these complications, PHT remains the most important cause of morbidity and mortality in patients with cirrhosis. Over the years, it has become clear that a decrease in portal pressure is not only protective against the risk of variceal (re)bleeding but is also associated with a lower long-term risk of developing complications and an improved long-term survival. A milestone in therapy was the introduction of non-selective beta-blockers for the prevention of bleeding and rebleeding of gastro-esophageal varices. However, in practice, less than half the patients under beta-blockade are protected from these risks, supporting the overall demand for innovation and expansion of our therapeutic armamentarium. Recent advances in the knowledge of the pathophysiology of cirrhotic PHT have directed future therapy towards the increased intrahepatic vascular resistance, which, in part, is determined by an increased hepatic vascular tone. This increased vasculogenic component provides the rationale for the potential use of therapies aimed at increasing intrahepatic vasorelaxing capacity via gene therapy, liver-selective nitric oxide donors and statines on the one hand, and at antagonizing excessive intrahepatic vasoconstrictor force through the use of endothelin antagonists, angiotensin blockers, alpha(1) adrenergic antagonists or combined alpha(1)- and non-selective beta-blockers or somatostatin analogues on the other. The focus of this review is to give an update on the pathophysiology of PHT in order to elucidate these potential novel strategies subsequently. PMID- 16343057 TI - Microarray technology in the study of obesity and non-alcoholic fatty liver disease. AB - The recent development of high-throughput gene expression technology permits simultaneous investigation of thousands of genes, providing a snapshot of the transcription state of diseased tissue. Microarray-based expression profiling is well suited to investigate the molecular basis of complex diseases such as obesity and chronic liver disease. With the help of microarray technology, functional genomics will surely advance our understanding of these diseases, and lead to more effective, targeted interventions that lack the toxicity of many conventional treatments. Despite their tremendous potential, microarray studies are subject to potential flaws in experimental design, experimental techniques, data analysis, and data interpretation. Besides the technical issues, the most important challenge is to develop integrative databases that combine gene expression data with the clinical data. Over the next few years, advances in technology and refinements in study design and data analysis will make clinically relevant translational research even more engaging and productive. PMID- 16343058 TI - Therapeutic implications of hepatitis B virus genotypes. AB - BACKGROUND/AIMS: Hepatitis B virus (HBV) is a global health problem. In addition to the implementation of universal hepatitis B vaccination, effective and individualized treatment of chronic hepatitis B to prevent progression into end stage liver diseases and hepatocellular carcinoma is still needed. HBV has been designated eight genotypes (A-H) based on genome sequence divergence. The epidemiology of HBV genotypes and their implications on the responses to antiviral therapy have become increasingly recognized in both Asian and Western countries. METHODS: Published data are thus reviewed. RESULTS: Each genotype has its distinct geographical and ethnic distribution. Genotypes A and D occur frequently in Africa, Europe, and India, while genotypes B and C are prevalent in Asia. Genotype E is restricted to West Africa, and genotype F is found in Central and South America. The distribution of genotypes G and H is less clear. Accumulating evidence indicates a better sustained response to conventional interferon in patients with genotype B than those with C, and in patients with genotype A than those with D. In contrast, conflicting results exist regarding the response to pegylated interferon. On the other hand, the therapeutic responses to nucleoside/nucleotide analogues are comparable among patients with different HBV genotypes. The impact of HBV subgenotypes, mixed genotype infections, and recombinants of different genotypes on the response to antiviral treatments awaits further examinations. CONCLUSION: Remarkable clinical and pathogenic differences do exist among HBV genotypes; however, researches on molecular and virologic mechanisms underlying the clinical phenotypes of different HBV genotypes are urgently needed. PMID- 16343059 TI - Liver allograft radiotherapy to treat rejection in children: efficacy in orthotopic liver transplantation and long-term safety. AB - BACKGROUND: We studied, retrospectively, the efficacy to control rejection and long-term safety of liver allograft radiotherapy (RT) performed in 14 children. Long-term safety data were collected with the prospect of possible use of RT in liver cell transplantation (LCT). METHODS: Immune suppression included cyclosporine, azathioprine and prednisone. In case of intractable rejection, low dose allograft RT was administered daily for 3 days, and short-term efficacy was evaluated by liver enzyme assays and histology. The long-term outcome was compared with that of 122 patients undergone transplantation and who had similar treatment, but no RT. RESULTS: Survival at 15 years was 71.4% vs 69.7% in the comparison group. In the RT group, rejection control was complete in six of 14 children and partial in two, all being alive and well 14-18 years later. Ten of 14 children had follow-up biopsy. Six children had normal histology and four had mild unspecific fibrosis. The long-term follow-up biopsy in the comparison group showed fibrosis in 42 of 85 children. The incidence of complications was similar in both groups. CONCLUSIONS: This series shows that, such a RT regimen appeared to be efficient and safe as a rescue treatment for acute rejection. Provided that further investigations in animal models show a certain benefit of low-dose irradiation around LCT, such a regimen could be proposed in human liver cell transplant programmes. PMID- 16343060 TI - Hepatitis C-associated autoimmunity in patients coinfected with HIV. AB - BACKGROUND: Hepatitis C virus (HCV) infection is associated with multiple extrahepatic manifestations. It is unclear to what extent extrahepatic manifestations occur in HIV/HCV coinfection. METHODS: We prospectively assessed cross-sectional frequencies of autoimmune manifestations in HIV/HCV-coinfected patients (n=98), HIV-mono-infected (n=45) and HCV-mono-infected patients (n=78). Diagnostic vasculitis scores, HCV and HIV loads, CD4 cell counts, thyroid-, cardiolipin-, non-organ-specific tissue antibodies (nuclear, smooth muscle, anti liver-kidney-microsome, neutrophil-cytoplasmic) and cryoglobulins were determined. RESULTS: Synergistic effects of HCV and HIV infection were observed with respect to the prevalence of antibodies against thyroglobulin (HCV infection 15.4%, HIV infection 8.8%, HIV/HCV coinfection 30.6%; P<0.001) and cardiolipin antibodies (HCV infection 9.0%, HIV infection 31%, HIV/HCV coinfection 46%; P<0.001). Cryoglobulinemia type III, was significantly associated with HCV infection (HCV, 25.6%; HIV/HCV, 20.4%) but not with HIV infection (4.4%, P<0.05). Rheumatoid factor was commonly detected in patients with HCV infection (48%), but occurred considerably less frequently in patients with HIV infection (4.4%) or HIV/HCV coinfection (9.5%, P<0.01). CONCLUSION: HIV coinfection appears to differentially modulate the frequency of HCV-related autoimmunity. However, autoimmunity is rarely accompanied by clinical manifestations. PMID- 16343061 TI - Association between human leukocytes antigen alleles and chronic hepatitis C virus infection in the Korean population. AB - BACKGROUND/AIM: Recent data have shown that the clinical outcome of hepatitis C virus (HCV) infection may be influenced by the host genetic factor. The aim of this study was to investigate whether particular human leukocytes antigen (HLA) molecules are associated with the susceptibility to HCV infection in the Korean population. METHODS: One hundred and thirty-seven patients with chronic HCV infection and 206 normal individuals were examined for HLA class I and II molecules. RESULTS: In class I antigens, the frequencies of HLA-A3 (relative risk (RR)=3.5, P<0.04), HLA-B35 (RR=2.0, P<0.03), and HLA-B46 (RR=2.5, P<0.02) significantly increased in chronic HCV carriers compared with the controls. The frequencies of DRB1*0803, DQB1*0601 and DQB1*0604 were significantly higher in chronic HCV carriers than in controls (RR=2.5, P<0.005; RR=1.8, P<0.05; RR=1.9, P<0.04, respectively). On the other hand, the frequencies of DRB1*0301, DQA1*0501 and DQB1*0201 were significantly lower in chronic HCV carriers than in normal controls (RR=0.2, P<0.03; RR=0.4, P<0.004; RR=0.5, P<0.02, respectively). The haplotype DRB1*0803-DQB1*0601 significantly increased (RR=2.5, P<0.02) while the DQA1*0501-DQB1*0201 significantly decreased (RR=0.2, P<0.03) in chronic HCV carriers compared with normal controls. In stratification analysis to investigate the interrelationships among the associated alleles, DRB1*0803 and DQB1*0601 were associated with HLA-B46, particularly in patients with chronic HCV carriers. CONCLUSIONS: These results suggest that particular HLA alleles may have an influence on chronic HCV infection as a host genetic factor in the Korean population. PMID- 16343062 TI - Low fibrosis progression of recurrent hepatitis C in apolipoprotein E epsilon4 carriers: relationship with the blood lipid profile. AB - BACKGROUND: The histological outcome of chronic hepatitis C is better among carriers of the apolipoprotein E (ApoE) epsilon4 allele, for reasons unknown. The orthotopic liver transplantation (OLT) setting allows to separate the role played by liver-derived ApoE (graft) from ApoE of different origin (recipient). PATIENTS AND METHODS: Forty-six OLT recipients with recurrent hepatitis C were studied. Grafts and recipients were genotyped for ApoE. In a follow-up extending up to 4 years, the serum triglycerides-to-cholesterol ratio (T/C ratio) was measured 1 year after OLT, whereas fibrosis progression was assessed yearly and expressed as fibrosis units/month (FU/mo). RESULTS: A T/C ratio < or =0.75 was observed in 13/15 cases in which both donor and recipient were epsilon4 carriers, 10/19 cases in which epsilon4 alleles were of exclusive recipient's origin and 5/12 cases in which epsilon4 alleles were of exclusive donor's origin or absent (P<0.02). One year after OLT, a fibrosis progression < or =0.100 FU/mo was associated with a low T/C ratio (24/34 vs. 4/12, P<0.05). An Ishak staging score >2 was reached later by male recipients who were epsilon4 carriers (P<0.002). CONCLUSIONS: Recipient's carriage of ApoE epsilon4 affects fibrosis progression of recurrent hepatitis C through gender-specific mechanisms, associated with a peculiar, ApoE associated, lipid profile. PMID- 16343063 TI - Evaluation of clinical usefulness of second-generation HCV core antigen assay: comparison with COBAS AMPLICOR HCV MONITOR assay version 2.0. AB - BACKGROUND: Hepatitis C virus (HCV) is an important etiologic agent for chronic liver diseases. METHODS: The aim of this study was to evaluate the clinical usefulness of second-generation HCV core antigen assay by comparing the results of the assay with those of the COBAS AMPLICOR HCV MONITOR version 2.0 (COBAS v2.0). RESULTS: HCV core antigen was detectable by this assay in 142/149 (95.3%) of serotype 1 (3821+/-322 fmol/l; mean+/-SD), in 56/58 (96.6%) of serotype 2 (2589+/-449 fmol/l), and in 6/6 (100%) of serotypes 1+2 (1240+/-548 fmol/l). The HCV core antigen levels measured by this assay correlated well with the HCV RNA levels by COBAS v2.0 (r=0.848, P<0.0001). In relation to the outcome of interferon monotherapy, the pretreatment HCV core antigen levels of sustained and non-sustained virological responders were 659+/-189 and 4904+/-376 fmol/l in serotype 1, 1993+/-740 and 3145+/-519 fmol/l in serotype 2. The cutoff values with the best accuracy for HCV core Ag levels to discriminate between sustained and non-sustained virological response were 699 fmol/l for serotype 1 and 292 fmol/l for serotype 2, respectively, by receiver operating characteristic curve analysis. CONCLUSION: This new assay was considered to be useful in evaluating the HCV levels in patients with chronic hepatitis C. PMID- 16343064 TI - Advances in digital quantification technique enhance discrimination between mild and advanced liver fibrosis in chronic hepatitis C. AB - BACKGROUND AND AIMS: The necessity of liver biopsy for staging fibrosis and its quantification in patients with chronic hepatitis C (CHC) remains controversial. Semiquantitative scoring of fibrosis is considered more subjective and less objective than digital quantification by image analysis. However, measurement of fibrosis using digital image analysis is thought to be less reliable in determining early stage fibrosis as compared with advanced fibrosis or cirrhosis. Our aims were to correlate all Ishak stages of fibrosis (0-6) with fibrosis percentage (%) using computerized digital image analysis, and thereby seek to improve discrimination between varying levels of liver fibrosis. METHODS: Fibrosis % data were obtained by image analysis on 164 trichrome-stained liver biopsies from untreated patients with CHC, representing all Ishak stages of fibrosis. RESULTS: Digital analysis of fibrosis % was highly correlated with Ishak scores of fibrosis (Kendall's tau-beta=0.86, P<0.001). Receiver-operator characteristic curves showed reliable discriminative capability of our digital image measurement of fibrosis when compared with semiquantitative assessments of fibrosis. Excellent interobserver reliability was found. CONCLUSIONS: Recent advances in digital quantification of fibrosis have resulted in improved discrimination between the varying stages of liver fibrosis, including mild fibrosis. This method is reproducible, can detect early as well as advanced fibrosis or cirrhosis, may prove to be the best assessment of mild fibrosis, and may be more precise than semiquantitative estimation of changes for monitoring fibrosis progression or regression during clinical therapeutic trials. PMID- 16343065 TI - Noninvasive assessment of liver steatosis, fibrosis and inflammation in chronic hepatitis C virus infection. AB - BACKGROUND: Duplex-Doppler ultrasound is a noninvasive method for the assessment of hepatic hemodynamics beyond conventional gray-scale imaging. The clinical value of the method for the grading and staging of chronic hepatitis C virus (HCV) infection and the prediction of hepatic steatosis still has to be determined. This study aimed to compare Duplex-Doppler and ultrasound with the histologic staging and the estimation of hepatic steatosis in chronic HCV infection. PATIENTS AND METHODS: One hundred and nineteen consecutive patients with chronic HCV infection underwent both liver biopsy and ultrasound with Duplex Doppler. Maximum portal venous blood flow velocity, portal venous flow undulation, hepatic venous flow pattern and spleen size were assessed and compared with histologic findings. Histologic grading and staging was performed according to the modified HAI and hepatic steatosis was estimated. RESULTS: Doppler ultrasound was unable to discriminate between different degrees of fibrosis. Sensitivity/specificity of portal venous flow and undulations for the diagnosis of hepatic cirrhosis was 74.5%/53% and 76.5%/100%. The PPV and NPV of reduced undulations was 100% and 96.2%. Mono- or biphasic hepatic venous flow indicated advanced hepatic steatosis (sensitivity 88.2%, specificity 74.5%, PPV 36.6%, NPV 97.5%). Spleen size was significantly enlarged both in patients with cirrhosis and steatosis. CONCLUSIONS: Although Duplex-Doppler of the portal and hepatic veins is not a substitute for histologic grading and staging, portal vein undulations can predict liver cirrhosis with considerable accuracy. Moreover, triphasic patterns of hepatic venous flow virtually exclude significant fatty liver disease. Additional studies should perform intraindividual follow-up investigations to further define the role of Duplex-Doppler ultrasound in chronic HCV infection. PMID- 16343066 TI - Simplified method of hepatic fibrosis quantification: design of a new morphometric analysis application. AB - INTRODUCTION: Semiquantitative evaluation of liver specimens is considered the standard method for measuring fibrosis; however, these systems lack the precision of a quantitative technique. METHODS: We developed an image analysis application (FibroXact) that automates and simplifies color segmentation. Trichrome slides were scanned and the program was used to evaluate each pixel based upon hue, lightness, and saturation values. Percent fibrosis was automatically calculated after thresholding. RESULTS: Linear regression demonstrated a strong relationship between the FibroXact result and the semiquantitative result (P<0.001). CONCLUSION: We have developed an image analysis application to quantify liver fibrosis. Correlation between our results and a standard semiquantitative system was demonstrated. PMID- 16343067 TI - Study of hepatitis B (HB) vaccine non-responsiveness among health care workers from an endemic area (Taiwan). AB - OBJECTIVE: To evaluate the etiology of non-responsiveness to hepatitis B (HB) vaccination in adults from an endemic area. METHODS: A total of 250 subjects who were HBsAg negative and anti-HBs<10 mIU/ml received three-dose HB-vaccine series. Anti-HBs 'negative' was defined as a level<1.5 mIU/ml. 'Weakly' positive was defined as 1.5-10 mIU/ml at pre-vaccination testing. Anti-HBs response was defined as a level >10 mIU/ml at post-vaccination testing. Among non-responders who were anti-HBc positive, serum anti-HBe and hepatitis B virus (HBV) DNA were tested. RESULTS: Three variables were associated with non-responsiveness by univariate analysis: anti-HBc positive, male gender, and age >40 years. Multivariate analysis additionally showed that anti-HBs negative was associated with non-responsiveness. Among 23 non-responders in anti-HBc positive subjects, post-vaccination serum was available in 16 subjects. HBV-DNA in all subjects was under detectable level by PCR assay. Anti-HBe positive were found in 13 of 16 subjects and were assumed to be occult HBV infection. CONCLUSION: Male gender, age >40 years and anti-HBc positive are associated with non-responsiveness to HB vaccination. Most of non-responders among anti-HBc positive subjects were assumed to be occult HBV infection. Subjects with weakly positive anti-HBs were associated with responsiveness which may be the effect of immune memory. PMID- 16343068 TI - Prevention of hepatitis B virus infection from hepatitis B core antibody-positive donor graft using hepatitis B immune globulin and lamivudine in living donor liver transplantation. AB - BACKGROUND: Hepatic grafts from hepatitis B surface antigen-negative and anti core antibody (HBcAb)-positive donors have been shown to transmit hepatitis B virus (HBV) infection. Recently, it has been reported that combined hepatitis B immune globulin (HBIG) and lamivudine therapy is effective in the prevention of hepatitis B recurrence after living donor liver transplantation (LDLT). In this report, we assessed the efficacy of combined HBIG and lamivudine therapy in preventing HBV transmission by graft with HBcAb-positive donors. METHODS: We studied 22 patients who had undergone LDLT with allografts from HBcAb-positive living donors at Gunma University Hospital and Kyushu University Hospital. Long term combined HBIG and lamivudine therapy were administrated to all recipients. Serum samples from the donor and recipient were tested for HBcAb, HBV DNA, and hepatitis B surface antibody. Liver biopsies from grafts were tested for HBV DNA. RESULTS: All recipients were HBcAb negative before LDLT. All of the donor livers were HBV DNA positive at the time of LDLT. All of the recipients had HBsAb titers greater than 300 mIU/ml 4 weeks after LDLT, and remained 100 mIU/ml thereafter. None of the recipients have become infected with HBV with a follow-up of 25-86 months. CONCLUSIONS: Perioperative combined HBIG and lamivudine therapy can prevent HBV infection in recipients who receive liver grafts from HBcAb-positive donors. PMID- 16343069 TI - Tumor necrosis factor gene polymorphisms and clearance or progression of hepatitis B virus infection. AB - OBJECTIVES: We evaluated the influence of tumor necrosis factor-alpha (TNF-alpha) promoter gene polymorphisms on clearance of hepatitis B virus (HBV) and outcome of HBV chronic hepatitis. METHODS: Four TNF-alpha promoter polymorphisms (T 1031C, C-863A, G-308A, and G-238A) were evaluated by direct sequencing in 184 chronic HBV carriers hepatitis B surface antigen (HBsAg) positive and 96 controls with documented sero-clearance (HBsAg negativity, positivity for anti-HBs and anti-HBc IgG). Frequencies of single-nucleotide polymorphisms (SNPs) and haplotypes in the control group were compared with those of the chronic carrier group and with clinically defined subgroups of the latter: asymptomatic carriers, patients with compensated hepatitis, decompensated cirrhotics, and patients with hepatocellular carcinoma. Furthermore, subgroups of chronic carriers were compared among them. RESULTS: In the chronic carrier group, the -308 G allele was more frequent in those with a family history of HBV infection (96% vs 88% of those with non-familial transmission). The G/G genotype at position -308 was found in all chronic carriers with decompensated cirrhosis but in only 78% of controls (P=0.01) and was more frequent in decompensated cirrhotics than in the other subgroups. The distribution of TNF-alpha gene polymorphisms in the carrier group was not significantly different from that in the sero-clearance control group. TNF-alpha SNPs at positions -1031/-863 and -863/-238 were in linkage disequilibrium. The TCGG haplotype (-T1031, -C863, -G308, -G238) was significantly associated with end-stage liver disease. CONCLUSION: The TNF-alpha promoter polymorphisms do not appear to be determinant of HBV sero-clearance in southern Italians. The genotype -308G/G and haplotype TCGG are associated with an unfavorable prognosis in patients with chronic HBV infection. PMID- 16343070 TI - Influence of HLA compatibility and lymphocyte cross-matching on acute cellular rejection following living donor adult liver transplantation. AB - BACKGROUND: Reports on the relevance of immunogenetic factors in living donor adult liver transplantation (LDALT) are often conflicting or inconclusive. We therefore investigated the human leukocyte antigen (HLA) mismatches, lymphocyte crossmatch positivity, and the reactivity in mixed lymphocyte culture (MLC) in a series of LDALT. METHODS: A total of 104 LDALT patients were studied. The minimum follow-up was 12 months, and the graft survival rates were assessed. The incidence of the most common complications was analyzed. And the influence of HLA, the flow cytometric analysis findings, enhanced cytotoxic cross-matching and MLC on graft survival, and acute rejection was also investigated. RESULTS: As a result, 96 negative cross-matching and eight positive cross-matching cases were identified. Positive cytotoxic cross-matching had a significant effect on graft survival (P<0.05), while flow cytometric cross-matching also had an additional effect on acute rejection (P<0.05). The MLC of the patients with three HLA mismatches was significantly higher than the MLC of patients with zero HLA mismatches. The incidence of acute cellular rejection (ACR) was higher in the patients with three mismatches than in the other patients, and moderate rejection only occurred in the patients with three mismatches. CONCLUSION: HLA mismatching was not statistically associated with the overall graft survival after LDALT. The graft failure rates were higher in the positive cross-matching cases and therefore a strong immuosuppressant might be needed for positive cross-matching cases. PMID- 16343071 TI - Local radiotherapy as a complement to incomplete transcatheter arterial chemoembolization in locally advanced hepatocellular carcinoma. AB - PURPOSE: In order to determine the effect of additional radiotherapy (RT) after an incomplete transcatheter arterial chemoembolization (TACE) in an unresectable hepatocellular carcinoma (HCC), the treatment results of patients receiving TACE plus RT were analyzed and compared with those treated with TACE alone. MATERIALS AND METHODS: One hundred and five patients with an unresectable HCC were treated with TACE from January 1992 to December 2002. In 73 of these patients, the TACE was incomplete. Among them, TACE was repeatedly performed in 35 patients (TACE group), and the remaining 38 patients were also treated with local RT (TACERT group). The patients were either in stage III or IVa, Eastern Cooperative Oncology Group 2 or less, and Child-Pugh class A or B. The average frequency of TACE prior to RT was 2 and the RT was started within 7-10 days after the TACE. RESULTS: The 2-year survival rate was significantly higher in the TACERT than in the TACE group (36.8 % vs. 14.3%, P=0.001). According to the tumor size, the 2 year survival rates in the TACERT and TACE groups were 63% vs. 42% in 5-7 cm (P=0.22), 50% vs. 0% in 8-10 cm (P=0.03), and 17% vs. 0% in larger than 10 cm (P=0.0002) respectively. CONCLUSION: There was a significantly improved survival rate in the TACERT group of unresectable HCC patients than in the TACE group, particularly in case of tumors > or =8 cm in diameter. Therefore, RT in addition to TACE is strongly recommended for patients with an unresectable HCC. PMID- 16343072 TI - Usefulness of urine strip test in the rapid diagnosis of spontaneous bacterial peritonitis. AB - PURPOSE: Rapid and accurate diagnosis of spontaneous bacterial peritonitis (SBP) is mandatory for timely treatment in cirrhotic patients. The purpose of this study was to assess the usefulness of two different reagent strips, the UriSCAN and the Multistix10SG, for the rapid bedside diagnosis of SBP. METHODS: A total of 75 paracenteses in 53 cirrhotic patients with ascites were performed. All ascitic fluid was analyzed with the two reagent strips, and compared with the manual cell count with differential and ascitic fluid culture. SBP was defined as an ascitic polymorphonuclear cell count > or =250/mm3. RESULTS: SBP was diagnosed in 18 of the 75 samples. If we considered the positive UriSCAN result of 2 or more, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were all 100%. When we considered the positive UriSCAN result of 3, the sensitivity, specificity, PPV, and NPV were 67%, 100%, 100%, and 89%, respectively. When we considered the positive Multistix10SG result of 3, the sensitivity, specificity, PPV, and NPV were 50%, 100%, 100%, and 87%, respectively. CONCLUSION: Urine reagent strip might be useful for rapid and accurate diagnosis of SBP in cirrhotic patients with ascites. PMID- 16343073 TI - Promoter methylation and loss of coding exons of the fragile histidine triad (FHIT) gene in intrahepatic cholangiocarcinomas. AB - AIMS: About 10-30% of primary liver cancers represent intrahepatic cholangiocarcinomas (IHCC). Since chromosomal losses of 3p are detectable in about 40% of cholangiocarcinomas our study aimed at the identification of mechanisms leading to functional deletion of tumor suppressor genes in this region. Our efforts focussed on genomic losses and epigenetic inactivation of two tumor suppressor genes, the fragile histidine triad (FHIT) and the ras association domain family 1 (RASSF1A) genes, both located on the short arm of chromosome 3. METHODS: Methylation-specific PCR (MSP) and combined bisulfite dependent restriction analysis (COBRA) were applied to detect epigenetic silencing of gene promoters. Genomic duplex PCR was used to identify exon losses of the FHIT gene. Nineteen paraffin-embedded samples of intrahepatic cholangiocarcinomas were studied. RESULTS: Here we report for the first time that in addition to frequent losses of the exons 5 and 6, hypermethylation of the FHIT promoter occured in a significant portion of IHCC. Methylation specific PCR (MSP) detected epigenetic inactivation of the FHIT/FRA3B locus in 8 of 19 (42%) cases. Combined bisulfite restriction analysis (COBRA) revealed that high levels of methylated FHIT promoter sequences were present in 6 of the 8 methylation positive samples. In agreement with previous reports MSP identified hypermethylation of the RASSF1A gene in 13 of 19 (68%) IHCC specimens examined. CONCLUSIONS: Epigenetic silencing of the FHIT tumor suppressor gene is a novel inactivation mechanism to be considered in the development of intrahepatic cholangiocarcinomas. However, a statistically significant inverse correlation between K-Ras activation and RASSF1A inactivation was not found. PMID- 16343074 TI - Methylation status of p14ARF, p15INK4b, and p16INK4a genes in human hepatocellular carcinoma. AB - BACKGROUND: The INK4 locus consisting of three genes involved in the regulation of cell cycle, p16INK4a, p15INK4b, and p14ARF is often disrupted in human neoplasms. METHODS: We analyzed the promoter methylation of each gene by methylation-specific PCR in hepatocellular carcinoma (HCC). RESULTS: The methylation of p16INK4a, p15INK4b, and p14ARF was found to occur in 27 (69.2%), seven (17.9%), and none out of 39 HCC tumors, respectively. Regarding corresponding nontumorous liver tissues, the promoter regions of p16INK4a, p15INK4b, and p14ARF were methylated in three (17.6%), three (17.6%), and none out of 17 samples, respectively. Analysis of mRNA expression revealed that loss of p16INK4a expression was frequently observed in HCC. In contrast, transcripts of p14ARF and p15INK4b were detected in 16 (88.9%) and 16 (88.9%) of 18 tumors, respectively. CONCLUSIONS: The frequent loss of transcription of p16INK4a with promoter methylation not only in the advanced but also in the early stages of HCC suggests that the epigenetic alteration of p16INK4a promoter is likely to be involved in hepatocarcinogenesis. Together with the result of RT-PCR analysis, the role of aberrant methylation of p14ARF or p15INK4a promoter in hepatocarcinogenesis is thought to be limited. PMID- 16343075 TI - Evaluation of liver parenchymal pressure and portal endothelium damage during radio frequency ablation in an in vivo porcine model. AB - BACKGROUND/AIMS: We previously developed a multi-step, incremental expansion method (multi-step method) for radio frequency ablation (RFA) in vitro, which prevented increases in pressure and reduced the ablation time as compared with other methods. In this study, we evaluated liver parenchymal pressure and portal endothelium damage during RFA with different devices and protocols in an in vivo porcine model. METHOD: Nine healthy female pigs were anaesthetized. RFA was performed with two different devices and protocols; one involved the use of a LeVeen needle with a single-step full expansion method or a multi-step method, and the other used a cool-tip needle with 40 or 60 W power. We measured the pressure in the liver parenchyma and the gallbladder during RFA. We also evaluated portal endothelium damage by NADH staining. RESULTS: The multi-step method with the LeVeen electrode resulted in the lowest parenchymal and intra gallbladder pressures (multi-step method2500 g). Our data failed to show a possible link between hospital admission with SS and development of atopic disease. PMID- 16343084 TI - Antibody levels to Bordetella pertussis in 10-yr-old children with atopy and atopic asthma. AB - Suboptimal immune responses to vaccination have been suggested among atopic infants. The aim of this study was to assess the influence of atopy and atopic asthma on the humoral response to Bordetella pertussis vaccination. Immunoglobulin (Ig)G and IgA specific antibodies were measured by enzyme linked immunosorbent assay in 102, 10-yr-old atopic children (66 of them also being asthmatics) and compared with 76 non-atopic and 53 non-atopic non-asthmatic controls of similar age. The levels of antibodies and the percentage of positives to B. pertussis were comparable in all groups. Children with a very high total serum immunoglobulin (Ig)E (Percentile (Pct) > 90th) showed higher (p = 0.01) IgG pertussis antibodies than children with very low serum IgE (Pct < 10th). In conclusion, we found normal pertussis antibody levels in atopic and in atopic asthmatic children in late childhood, thus overriding any possible suboptimal response during infancy. PMID- 16343085 TI - Feeding a soy formula to children with cow's milk allergy: the development of immunoglobulin E-mediated allergy to soy and peanuts. AB - Peanut allergy has been associated with the intake of soy milk or a soy formula. We studied the development of immunoglobulin E antibodies specific to soy and peanuts and of allergic reactions caused by peanuts, in children with confirmed cow's milk (CM) allergy fed either a soy formula or an extensively hydrolyzed formula (EHF). One hundred and seventy infants with documented CM allergy (CMA) were randomly assigned to receive either a soy formula or an EHF. The children were followed to the age of 4 yr. Peanut-specific immunoglobulin E was measured at the age of 4. A detailed history of the occurrence of allergic reactions caused by peanuts was recorded by the parents. Soy-specific immunoglobulin E antibodies were measured at the time of diagnosis and at the ages of 1, 2 and 4 yr. Immunoglobulin E antibodies to soy (> or =0.35 kU/l) were found in 22 of 70 children fed the soy formula, and in 14 of 70 of the children fed the EHF (p = 0.082). In an open challenge with soy at the age of 4, no immediate reactions were observed. One of 72 children from the soy group had a delayed reaction. immunoglobulin E antibodies to peanuts (> or =0.35 kU/l) were found in 21 of 70 children fed the soy formula and 17 of 69 infants fed the EHF (p = 0.717). The incidence of reported peanut allergy in the soy group was two of 72 (3%) and four of 76 (5%) in the EHF group (p = 0.68). Development of immunoglobulin E associated allergy to soy and peanuts was rare in our study group of milk allergic children. The use of a soy formula during the first 2 yr of life did not increase the risk of development of peanut-specific immunoglobulin E antibodies or of clinical peanut allergy. PMID- 16343086 TI - Comparison of the allergenicity of Actinidia deliciosa (kiwi fruit) and Actinidia chinensis (gold kiwi). AB - Actinidia chinensis (gold kiwi) is a newly available fruit which has been shown to have in vitro immunoglobulin E (IgE) cross-reactivity with green kiwi. This is the first study to investigate clinical reactivity of gold kiwi. Five patients clinically allergic to green kiwi were investigated by skin test and double-blind placebo controlled food challenge (DBPCFC) with gold kiwi fruit. IgE-binding patterns of individual sera from the five challenged patients and a pool of sera from a further nine patients with kiwi allergy were compared in the two fruits by Western blotting. Cross reactivity of proteins in the two fruits was assessed by inhibition of immunoblots and by IgE enzyme-linked immunosorbent assay (ELISA) inhibition. Four of the five patients had a positive DBPCFC to gold kiwi. Western blotting showed marked differences in the allergen patterns of green and gold kiwi. However, inhibition of the immunoblots and ELISA assay reveals extensive inhibition of IgE binding to proteins in each fruit by the alternative species. Gold kiwi fruit is allergenic and patients allergic to green kiwi are at risk of reacting to the gold kiwi fruit. Despite having different protein profiles and IgE-binding patterns, the two species have proteins that extensively cross inhibit the binding to IgE. PMID- 16343087 TI - Exercise-induced bronchoconstriction in children with atopic eczema. AB - Non-specific bronchial hyper-responsiveness has been reported in most of the eczematous children even in the absence of asthmatic symptoms. We therefore investigated the occurrence of exercise-induced bronchoconstriction (EIB) in children with atopic eczema (AE) and the predictors of EIB. Fifty-five children referred to the paediatric clinic for AE and a control group of 17 healthy children were recruited. They all carried out a physical examination and skin prick test (SPT) both to inhalant and food allergens, prior to the exercise challenge test. Their parents filled a questionnaire on atopic diseases. They underwent exercise challenge test that consisted in free running for 6 min. Spirometric measurements were carried out before running and till 11 min after. Exercise challenge test was positive in 13 (23%) children with AE. None of the children in the control group had a positive exercise challenge test [OR (95% CI) = 1.31 (1.13-1.51); p = 0.030]. Sixteen (29%) eczematous children had a history of EIB. Such history was not reliable for identifying children who had a positive exercise test. Twenty-nine (52%) children with AE had asthma. Allergic rhinitis affected 33 (60%) of eczematous children and allergic conjunctivitis 28 (50%). EIB was not related to any history of asthma, allergic rhinitis, allergic conjunctivitis, severity of eczema or SPT results. Our study shows that EIB is common in children with AE. Our data also indicate that screening by medical history and physical examination is not a sensitive marker of EIB. This may explain why EIB is often ignored in eczematous children. PMID- 16343088 TI - Occupational asthma in apprentice adolescent car painters. AB - Occupational asthma (OA) is one of the leading causes of pulmonary diseases and has been extensively studied in adults. Childhood employment, a significant problem in many developing countries, should be studied to determine and evaluate its effects on psychosocial and lung health. In order to investigate the presence of work-related asthma-like symptoms and OA in apprentice adolescent car painters, 72 adolescents between the ages of 15-20 yr studying in Vocational Training Centres of Ankara were investigated using questionnaire, pulmonary function test (PFT), serial peak expiratory flow (PEF) measurements and methacholine inhalation tests. As a control group, 72 adolescents studying in Industrial and Commercial Training Centres located in the same environment were investigated with questionnaire and PFT. Almost 50% of the study group had work related asthma-like symptoms for which occupational dermatitis history was predictive [odds ratio: 2.9 (1.026-8.13) (95% confidence interval)]. Seventeen of 22 with serial PEF measurements showed a variability of > or =20% and three (4.2%) of 12 tested with methacholine inhalation test had a PC20 < or = 8 mg/ml, which led to the diagnosis of OA. There was no statistically significant difference between study and control groups in terms of PFT. In conclusion, the high prevalence of work-related asthma-like symptoms among adolescent car painters clearly indicates the need for routine follow-up of adolescent workers for lung health. PMID- 16343089 TI - School-based asthma surveillance: a comparison of student and parental report. AB - This study addressed the comparability of data obtained from a student-based and parent-based asthma and respiratory health survey. Our goal was to ascertain whether there were meaningful and systematic differences in asthma classification based on symptom and diagnosis reports obtained separately from students and their parents. A brief, written survey, based on the International Study of Asthma and Allergy in Children questionnaire, was administered to 6th through 10th grade students in two schools in Oakland, CA, USA. Students who reported asthma-like indicators for the previous 12-month period were defined as positive and a more extensive questionnaire was mailed home to those parents. A more refined classification of asthma based on parent report of indicators was compared with student report. Forty-four percent of 1298 students were classified as positive for current asthma-like symptoms and 50% of parent surveys were returned. For the positive students with parent surveys, 59% were classified as 'probable' for asthma based on the parent survey. Overall, the agreement between parent and students' classification was 70%, and 83% for students with a parent report of physician diagnosis of asthma. Students who were discordant with parents for physician diagnosis of asthma were more likely to be male, and more likely to have a parent report of unscheduled Emergency Department visit for wheezing or trouble breathing. Findings indicated that with the exception of medication, students reported asthma indicators more frequently that parents, independent of classification. Student report of physician diagnosis with a 12 month report of an asthma symptom was determined to be a good indicator of probable current asthma. Inclusion of or reliance on a parental questionnaire is not likely to improve the reliability of a school-based asthma surveillance program in our population. PMID- 16343090 TI - Etiology of atopy in infancy: the KOALA Birth Cohort Study. AB - The aim of the KOALA Birth Cohort Study in the Netherlands is to identify factors that influence the clinical expression of atopic disease with a main focus on lifestyle (e.g., anthroposophy, vaccinations, antibiotics, dietary habits, breastfeeding and breast milk composition, intestinal microflora composition, infections during the first year of life, and gene-environment interaction). The recruitment of pregnant women started in October 2000. First, participants with 'conventional lifestyles' (n = 2343) were retrieved from an ongoing prospective cohort study (n = 7020) on pregnancy-related pelvic girdle pain. In addition, pregnant women (n = 491) with 'alternative lifestyles' with regard to child rearing practices, dietary habits (organic, vegetarian), vaccination schemes and/or use of antibiotics, were recruited through organic food shops, anthroposophic doctors and midwives, Steiner schools, and dedicated magazines. All participants were enrolled between 14 and 18 wk of gestation and completed an intake questionnaire on family history of atopy and infant care intentions. Documentation of other relevant variables started in the pregnant mother and covered the first and third trimester as well as early childhood by repeated questionnaires at 14-18, 30, and 34 wk of gestation and 3, 7, 12, and 24 months post-partum. A subgroup of participants, including both conventional and alternative lifestyles, was asked to consent to maternal blood sampling, breast milk and a faecal sample of the infant at 1 month post-partum, capillary blood at age 1 yr, venous blood and observation of manifestation of atopic dermatitis during home visits at the age of 2 yr (using the UK working party criteria and the severity scoring of atopic dermatitis index), and buccal swabs for DNA isolation from child-parent trios. From the start, ethical approval and informed consent procedures included gene-environment interaction studies. Follow-up at 3 and 7 months post-partum was completed with high response rates (respectively 90% and 88% in the conventional group, and 97% and 97% in the alternative group). The home visits at 2 yr of age will be completed in 2005. Preliminary results show that we have succeeded in recruiting a large population with various lifestyle choices with a fairly large contrast with regard to dietary habits (including organic foods, vegetarian diet), vaccination schemes and/or use of antibiotics. We have also been able to collect a large number of faecal samples (n = 1176) and capillary blood samples at age 1 yr (n = 956). Furthermore, a large proportion of the participants have consented with genetic studies. Mid 2006 we expect to report our first results on the relationship between the various exposures in early life and childhood atopy. An outline of the focus and design of the KOALA Birth Cohort Study is presented. PMID- 16343091 TI - Anterior chest wall protrusion as initial presentation of chronic granulomatous disease: a case report. AB - Chronic granulomatous disease (CGD) is a rare primary immunodeficiency disease characterized by recurrent life-threatening bacterial and fungal infections. We report a 4-month-old girl with CGD, was firstly presented with an anterior chest wall protrusion because of an aspergillosis mass. PMID- 16343092 TI - Male sex matters. PMID- 16343094 TI - Instruments used in measuring symptoms, health status and quality of life in hypothyroidism: a systematic qualitative review. AB - BACKGROUND: Symptoms of hypothyroidism are varied and nonspecific, thereby making clinical diagnosis impossible. Some patients report ongoing symptoms despite treatment. Measuring symptoms, health status and quality of life by using appropriate instruments, in addition to biochemical tests, is therefore vital to quantify disease severity and assess response to treatment. A number of instruments have been used in hypothyroidism, leading to lack of clarity about what exactly is being measured, and the rationale for the inclusion of particular measures is often uncertain. OBJECTIVES: To evaluate and compare instruments used in hypothyroidism in assessing patient-reported outcomes. DATA SOURCES AND STUDY SELECTION: All published literature on MEDLINE until March 2005 was searched using the keywords 'hypothyroidism', 'symptoms', 'quality of life' and 'health status'. Articles reporting controlled trials or instrument design and development were analysed to assess the instruments used and to ascertain whether they were measuring the stated outcome. DATA SYNTHESIS: Available tools to measure symptoms, health status and quality of life in diagnosis and management of hypothyroidism are discussed. We also describe how these tools have been described in published clinical trials along with the advantages and possible pitfalls of each of the commonly used tools. CONCLUSIONS: Some instruments reviewed have not been suited to the outcome being studied, thereby giving rise to questionable results. This review should be useful for clinicians, researchers and patient-support groups in understanding the principles behind these tools and the results obtained from their use in clinical practice, and in selecting measures for their own use. PMID- 16343093 TI - Endocrine regulation of bone turnover in men. PMID- 16343095 TI - Ectopic expression of vasopressin V1b and V2 receptors in the adrenal glands of familial ACTH-independent macronodular adrenal hyperplasia. AB - OBJECTIVE: ACTH-independent macronodular adrenal hyperplasia (AIMAH) is a rare and unusual cause of Cushing's syndrome, characterized by bilateral nodular adrenocortical hyperplasia and hypersecretion of cortisol. Familial AIMAH has rarely been reported. Recently, the aberrant expression of adrenal receptors for various ligands in AIMAH patients has become important in explaining the pathogenesis of AIMAH. In this study, we present the cases of two sisters who were affected with AIMAH. PATIENTS AND MEASUREMENTS: Two sisters, aged 46 and 58, respectively, at the time of diagnosis, were found to have Cushing's syndrome due to AIMAH. We performed provocation tests with various materials and reverse transcription polymerase chain reaction (RT-PCR) with their resected adrenal tissues to examine the hyper-responsiveness to steroidogenesis and the overexpression of related receptors, respectively. RESULTS: Provocation tests in vivo indicated that arginine vasopressin (AVP) promoted cortisol secretion through vasopressin V1a as well as V1b and V2 receptors. RT-PCR analysis revealed an abnormal cDNA expression of vasopressin V1b and V2 receptors, none of which is known to be normally expressed in the adrenal glands. CONCLUSION: The expression of ectopic vasopressin V1b and V2 receptors may be involved in the aetiology of AIMAH, at least in the case of the sibling patients presented here. PMID- 16343096 TI - Metformin administration and laparoscopic ovarian drilling improve ovarian response to clomiphene citrate (CC) in oligo-anovulatory CC-resistant women with polycystic ovary syndrome. AB - OBJECTIVE: To compare the efficacy of clomiphene citrate (CC) administration for ovulation induction in CC-resistant oligo-ovulatory women with polycystic ovary syndrome (PCOS) after metformin treatment or laparoscopic ovarian drilling (LOD). Design Prospective controlled study. PATIENTS: Twenty-eight overweight oligo amenorrhoeic women with PCOS who were still anovulatory after metformin administration (group A, n = 8) or LOD (group B, n = 20). METHODS: Treatment with CC 150 mg/day for 5 days from the third to the seventh day of a progesterone induced uterine bleeding for 6 months. MEASUREMENTS: The ovulation, pregnancy, abortion and live-birth rates were evaluated in each group. RESULTS: The subjects of groups A and B were studied for a total of 36 and 74 cycles, respectively. At the end of the study, no differences in ovulation [11/36 (30.6%) vs. 23/74 (31.1%); P = 0.96], pregnancy [6/36 (16.7%) vs. 12/74 (16.2%); P = 0.95], abortion [2/6 (33.3%) vs. 5/12 (41.7%); P = 1.00] and the live-birth [4/6 (66.7%) vs. 7/12 (58.3%); P = 1.00] rates were observed between groups A and B. CONCLUSION: Metformin administration as well as LOD improves the effectiveness of CC for ovulation induction in overweight infertile CC-resistant women with PCOS. PMID- 16343097 TI - Experience of prophylactic thyroidectomy in multiple endocrine neoplasia type 2A kindreds with RET codon 804 mutations. AB - OBJECTIVE AND DESIGN: Genetic screening in multiple endocrine neoplasia type 2 (MEN 2) has led to specific management guidelines based on genotype-phenotype analysis. However, there is controversy regarding the appropriate age for prophylactic thyroidectomy in families with mutations in codon 804 in exon 14 of the RET proto-oncogene, where medullary thyroid cancer (MTC) may not develop until adulthood. We prospectively studied two MEN 2A families, one with the V804L and the other with the V804M RET mutation, to report our experience of genetic and biochemical screening and prophylactic thyroidectomy. Family 1 is one of the largest MEN 2A families in the literature, where 22 prophylactic thyroidectomies have been performed. PATIENTS AND RESULTS: C-cell hyperplasia (CCH) was found in 23 out of 25 thyroidectomy specimens from family members of ages 5 years and upwards. MTC was found in 10 out of 18 adults of age 25 years upwards, including the family 2 proband, who was found to have MTC with lymph node metastases at age 28. Phaeochromocytoma was only observed in one patient, but six cases of histologically confirmed hyperparathyroidism were seen in family 1. CONCLUSION: We suggest that prophylactic thyroidectomy should not be delayed until adulthood in MEN 2A families carrying codon 804 RET mutations, but should be performed when there is CCH, before the development of MTC, as close as possible to age 6 years, which is the age of the youngest reported case of MTC in '804' families. PMID- 16343098 TI - Elevated late-night salivary cortisol levels in elderly male type 2 diabetic veterans. AB - OBJECTIVE: Late-night salivary cortisol (LNSC) is reportedly highly accurate for the diagnosis of Cushing's syndrome (CS). However, diagnostic thresholds for abnormal results are based on healthy, young populations and limited data are available on its use in elderly populations with chronic medical conditions. The purpose of this study was to evaluate LNSC levels in elderly male veterans with and without diabetes. DESIGN: Prospective evaluation of LNSC levels in male veterans. PATIENTS: One hundred and fifty-four participants with type 2 diabetes and 52 participants without diabetes. MEASUREMENTS: Participants underwent outpatient LNSC (2300 h) testing. Participants with elevated LNSC (> or = 4.3 nmol/l) underwent secondary testing, including 24-h urine free cortisol (24UFC, > 60 microg/day) and dexamethasone suppression testing (DST, serum cortisol > 50 nmol/l). Participants with positive secondary testing had a morning ACTH level analysed and either pituitary or adrenal imaging performed. RESULTS: One hundred and forty-one diabetics and 46 controls (mean age 61 years) returned samples (91% overall). Average LNSC levels (nmol/l) in diabetics were significantly higher than in nondiabetics [median (interquartile range): 2.6 (1.8-4.1) vs. 1.6 (1.0 2.0)] and in those aged > or = 60 compared to < 60 [2.7 (2.0-4.3) vs. 1.9 (1.4 2.9)] (P < 0.001 for both). Thirty-one participants required secondary testing. Seventy-nine per cent of participants who underwent secondary testing had normal 24UFC and DST. No cases of CS have been diagnosed to date. Increasing age [odds ratio (OR) 2.0 per decade], current diabetes mellitus (OR 4.4), and elevated blood pressure (OR 1.3 per 10 mmHg increase in systolic blood pressure) were associated with abnormal LNSC results (P < 0.05 for each). CONCLUSIONS: LNSC has been shown to be sensitive and specific in diagnosing CS in certain high-risk populations, primarily the young and middle-aged. The development of age- and comorbidity-adjusted thresholds may be warranted for LNSC testing in elderly subjects and in those with significant comorbidity. PMID- 16343099 TI - Does pravastatin increase chylomicron remnant catabolism in postmenopausal women with type 2 diabetes mellitus? AB - OBJECTIVE: We investigated the effects of pravastatin on chylomicron remnant catabolism measured with a 13C stable isotope breath test and plasma apolipoprotein (apo) B-48 and remnant-like particle (RLP)-cholesterol in postmenopausal women with type 2 diabetes mellitus. PATIENTS AND MEASUREMENTS: Nineteen postmenopausal women with type 2 diabetes were randomized to receive 40 mg/day pravastatin or no treatment for 6 weeks followed by a 2-week washout period, and crossed over for a further 6 weeks. Fractional catabolic rate (FCR) of a chylomicron remnant-like emulsion was determined from 13CO2 enrichment in the breath and plasma using isotope-ratio mass spectrometry and multicompartmental modelling. Plasma apo B-48 and RLP-cholesterol concentrations were also measured as static markers of chylomicron remnant metabolism. RESULTS: Pravastatin significantly reduced plasma concentrations of cholesterol (5.9 +/- 0.3 vs. 4.8 +/- 0.2 mmol/l; P < 0.001), low density lipoprotein (LDL)-cholesterol (3.5 +/- 0.2 vs. 2.6 +/- 0.2 mmol/l; P < 0.001), triglyceride (2.1 +/- 0.3 vs. 1.7 +/- 0.2 mmol/l; P = 0.017), non-high density lipoprotein (HDL)-cholesterol (4.4 +/- 0.3 vs. 3.3 +/- 0.2 mmol/l; P < 0.001), lathosterol/total cholesterol ratio (2.6 +/- 0.2 vs. 2.0 +/- 0.3, P = 0.035), apo B-100 (1.1 +/- 0.1 vs. 0.8 +/ 0.1 g/l; P = 0.001), apo B-48 (4.8 +/- 0.9 vs. 3.3 +/- 0.6 mg/l; P = 0.016), and RLP-cholesterol (31.4 +/- 8.2 vs. 18.6 +/- 4.6 mg/dl; P = 0.024). Pravastatin was also associated with an increase in sitosterol/total cholesterol ratio (2.8 +/- 0.3 vs. 3.1 +/- 0.3, P = 0.029). Chylomicron remnant-like emulsion catabolism was not, however, significantly altered by pravastatin estimated by either breath or plasma clearance measurements. CONCLUSIONS: In postmenopausal women, pravastatin decreases plasma concentrations of remnant lipoproteins by a mechanism that may relate chiefly to inhibition of remnant production, but this requires further evaluation. PMID- 16343100 TI - The significance of thyroid blood flow at the inferior thyroid artery as a predictor for early Graves' disease relapse. AB - OBJECTIVE: We investigated the clinical usefulness of thyroid blood-flow measurement in predicting relapse of Graves' disease (GD) in comparison with known risk factors for GD relapse. MEASUREMENT: Thyroid blood flow was measured in pulsed Doppler mode at the inferior thyroid artery (ITA), and the peak systolic velocity (PSV) calculated. PATIENTS: ITA-PSV was measured in euthyroid GD patients (n = 79) immediately before withdrawal of anti-thyroid drug (ATD) and in healthy subjects (n = 17). RESULTS: In the 79 euthyroid GD patients, the values of free triiodothyronine (FT3), TSH receptor autoantibody (TRAb), ITA-PSV and thyroid volume were significantly higher in the relapse group (n = 40) than in the nonrelapse group (n = 39) and the Youden index of ITA-PSV was significantly higher than that of FT3, TSH, TRAb and vascular endothelial growth factor (VEGF). CONCLUSION: ITA-PSV may assist in the prediction of early GD relapse after ATD withdrawal. PMID- 16343101 TI - Effects of Tai Chi and resistance training on cardiovascular risk factors in elderly Chinese subjects: a 12-month longitudinal, randomized, controlled intervention study. AB - BACKGROUND: Tai Chi is rapidly gaining in popularity, worldwide. This study was performed to assess its impact on cardiovascular risk factors in comparison with resistance training exercises in elderly Chinese subjects. METHODS: A total of 207 healthy elderly participants (65-74 years, 113/207 (55%) men) were randomly assigned to one of three intervention groups: (1) Tai Chi, three times/week for 1 h/session (n = 64); (2) resistance training exercise, three times/week for 1 h/session (n = 65); (3) usual level of physical activity control group (n = 78). Anthropometric measures, dual X-ray densitometry body composition, blood pressure, lipids, glycaemic and insulin sensitivity indices were measured at baseline and 12 months. Repeated-measures analysis of variance (anova) was used to assess the between-group changes using a last-observation-carried-forward intention-to-treat approach. RESULTS: A total of 180 (87.0%) subjects completed the study. No significant changes were identified in the Tai Chi group compared to the resistance training or control group. Of the primary outcomes, only the improvement in the insulin sensitivity index differed, being significantly greater in the resistance training than in the control group [mean difference 0.018 (95% confidence interval ( CI) 0.000-0.037) mmol glucose/min, P = 0.02), and tending to be greater than in the Tai Chi group (mean difference 0.019 (95% CI 0.000-0.038) mmol glucose/min, P < 0.06). CONCLUSION: Tai Chi had no significant effect on any measure compared to the controls, whereas resistance training improved the insulin sensitivity index in this 12-month study. PMID- 16343102 TI - Thyroid dysfunction and serum lipids: a community-based study. AB - OBJECTIVE: It is uncertain whether subclinical hypothyroidism (SCH) is associated with hypercholesterolaemia, particularly in subjects with SCH and serum TSH < or = 10 mU/l. DESIGN, PATIENTS AND MEASUREMENTS: Cross-sectional study of 2108 participants in a 1981 community health survey in Busselton, Western Australia. Serum total cholesterol and triglycerides were measured in all subjects and high density lipoprotein cholesterol (HDL-C) measured (and low density lipoprotein cholesterol (LDL-C) calculated) in a subgroup of 631 subjects at the time of the survey. In 2001, TSH and free T4 concentrations were measured on archived sera stored at -70 degrees C. Serum lipid concentrations in subjects with thyroid dysfunction and euthyroid subjects were compared using linear regression models. RESULTS: In the group as a whole, serum total cholesterol was higher in subjects with SCH (N = 119) than in euthyroid subjects (N = 1906) (mean +/- SD 6.3 +/- 1.3 mmol/l vs. 5.8 +/- 1.2 mmol/l, P < 0.001 unadjusted, P = 0.061 adjusted for age, age(2) and sex). Serum total cholesterol was similarly elevated in subjects with SCH and TSH < or = 10 mU/l (N = 89) (6.3 +/- 1.3 mmol/l, P < 0.001 unadjusted, P = 0.055 adjusted for age, age(2) and sex). In the subgroup analysis, LDL-C was higher in subjects with SCH (N = 30) than in euthyroid subjects (N = 580) (4.1 +/ 1.2 mmol/l vs. 3.5 +/- 1.0 mmol/l, P < 0.01 unadjusted, P = 0.024 adjusted for age, age(2) and sex). LDL-C was significantly increased in subjects with SCH and TSH < or = 10 mU/l (N = 23) (4.3 +/- 1.3 mmol/l, P < 0.001 unadjusted, P = 0.002 adjusted for age, age(2) and sex). CONCLUSION: SCH is associated with increased serum LDL-C concentrations, which is significant after adjustment for age, age(2) and sex. PMID- 16343103 TI - Multiple endocrine neoplasia 2A due to a unique C609S RET mutation presents with pheochromocytoma and reduced penetrance of medullary thyroid carcinoma. AB - OBJECTIVE: We have identified a large kindred with multiple endocrine neoplasia 2A (MEN 2A) due to a mutation at RET codon 609 that results in a cysteine to serine substitution, a mutation previously identified in only one case in the literature. We characterized the clinical phenotype of the kindred and the biochemical mechanism of this new mutation. PATIENTS AND DESIGN: The index case, a 42-year-old woman, presented with pheochromocytoma. We screened 29 family members for the presence of the mutation. Of the 15 mutation-positive family members, 11 agreed to undergo further evaluation by physical examination, calcium and pentagastrin-stimulated calcitonin levels, measurement of urinary metanephrines, adrenal imaging and serum calcium levels. Biochemical characterization of the mutation was by transient transfection of human neuroblastoma cells and Western blot analysis. RESULTS: This kindred demonstrated an inheritance pattern consistent with autosomal dominant pheochromocytoma. Strikingly, no clinically evident case of medullary thyroid cancer (MTC) was observed among mutation-positive family members. Thyroidectomy in six cases revealed C-cell hyperplasia in all and microscopic MTC in two cases. Transfection experiments using human neuroblastoma cells showed that the mutant RET, unlike the wild-type receptor, is constitutively phosphorylated in the absence of ligand, and thus resembles other previously characterized MEN 2A mutations. CONCLUSIONS: The identification of a new mutation causing a MEN 2A phenotype that features pheochromocytoma and the surprising absence of clinically apparent MTC has significant implications for carriers of this mutation and provides further insights into the genotype-phenotype correlation in MEN 2A. PMID- 16343104 TI - Apoptosis is reduced in the colonic mucosa of patients with acromegaly. AB - BACKGROUND: Patients with acromegaly have an increased risk of developing colonic tumours; reduced apoptosis is considered a leading mechanism in tumorigenesis. GH and IGF-1 decrease apoptosis in several cell lines including human colonic adenocarcinoma, but it is unknown whether epithelial cells of colonic mucosa of patients with acromegaly have reduced apoptosis. AIM: The aim of the study was to evaluate the degree of apoptosis in a cross-sectional study, in biopsy samples of colonic mucosa obtained from patients with acromegaly. PATIENTS AND METHODS: Eleven patients with active, untreated acromegaly (AcroUntr), 16 patients with acromegaly in remission (AcroRem) and 23 controls were enrolled in the study. Samples of colonic mucosa were obtained during colonoscopy; apoptosis was evaluated by either DNA fragmentation or terminal deoxynucleotidyl transferase assay. RESULTS: Apoptotic cells were 60.0 +/- 2.5% in samples of colonic mucosa of controls, 62.0 +/- 3.4% in those from patients with AcroRem (P = ns vs. controls), and 39.0 +/- 4.1% in those from patients with AcroUntr (P < 0.0001 vs. the other groups). Apoptosis was inversely related to serum IGF-I (r = 0.771, P < 0.001) or GH (r = 0.404, P = 0.05) levels and less to the estimated duration of disease (r = 0.384, P = 0.07). PPARgamma is considered to be a tumour suppressor gene the expression of which might be involved in colonic tumorigenesis. The expression of PPARgamma was lower in the colonic mucosa of patients with AcroUntr (2845 +/- 947 transcripts) than in that of controls (35 200 +/- 2450 transcripts) or AcroRem (29 547 +/- 3650 transcripts) (P < 0.005). The recovery of PPARgamma expression was associated with apoptosis in most cells. The lower degree of apoptosis in patients with AcroUntr was associated with a reduced expression of the antiapoptotic Bax protein. CONCLUSION: In conclusion, patients with AcroUntr have reduced apoptosis in colonic mucosa that is apparently reversed after acromegaly is cured. It is conceivable that reduced apoptosis may represent an early event in colonic tumorigenesis of patients with acromegaly. PMID- 16343105 TI - Is the anteroposterior and transverse diameter ratio of nonpalpable thyroid nodules a sonographic criteria for recommending fine-needle aspiration cytology? AB - BACKGROUND: As a consequence of the increasing application of ultrasound (US) technology, the detection of asymptomatic nonpalpable thyroid nodules has generally increased. The aim of our study was to assess if the anteroposterior and transverse diameter ratio of nonpalpable thyroid nodules (A/T) > or = 1 could be a sonographic criterion for recommending fine-needle aspiration cytology (FNAC). METHODS: From January 2002 to January 2004, 828 consecutive solid nonpalpable thyroid nodules were evaluated by ultrasonography, colour-Doppler and FNAC in our department. Cases were selected from 2217 patients, referred to our thyroid unit for US-guided FNAC from the greater Brescia area, an endemic zone for goitre. Entry criteria included the presence at US of a solid thyroid nodule that was nonpalpable at physical examination, euthyroid condition and no previous diagnosis of thyroid malignancy. All patients with suspicious or malignant cytology underwent surgery. RESULTS: One hundred and twenty-seven nodules with inadequate cytology were excluded from the study. Thyroid malignancy was observed in 67 (9.6%) nodules. At US, cancers presented a solid hypoechoic appearance in 79.1% of cases, blurred margins in 47.8%, microcalcification in 73.1%, intranodular vascular pattern in 56.7% and A/T > or = 1 in 83.6%. A hypoechoic appearance (OR 4.3), blurred margins (OR 2.6), microcalcification (OR 6.1), intranodular vascular pattern (OR 10.2) and A/T > or = 1 (OR 22.4) were independent risk factors of malignancy. CONCLUSIONS: A/T > or = 1 in conjunction with at least one other sonographic risk factor is able to detect the majority of carcinoma and, moreover, it limits the FNAC procedures to only 15.9% of all the nodules. PMID- 16343106 TI - Simultaneous Cushing's disease and tuberous sclerosis; a potential role for TSC in pituitary ontogeny. PMID- 16343107 TI - IL-13 and chromosome 5q31-q33: problems of identifying association within regions of linkage to Graves' disease. PMID- 16343108 TI - Gitelman's syndrome in late adulthood. PMID- 16343109 TI - Diabetic retinopathy, nephropathy and cardiovascular disease in a patient with GH gene deletion. PMID- 16343110 TI - Ex vivo comparison of two electronic apex locators with different scales and frequencies. AB - AIM: To compare ex vivo the accuracy of two impedance quotient apex locators with different scales and frequencies of the measuring circuit. METHODOLOGY: In each root of 193 extracted human teeth, electronic working length determination (ELD) was carried out with a newly constructed measuring unit. In all cases, ELD was performed using the apex locators Justy II (Hager & Werken, Duisburg, Germany) and Raypex (VDW, Munich, Germany) on the scale points (sp) 0/0.5/1 of each device. A Miller Needle reaching working length was fixed with composite. The corresponding sp and the differences to the other sp were recorded. After histological preparation of the apical region, the teeth were examined under a light microscope. The distances of the Miller Needle tips to the target intervals 'minor foramen-major foramen' and 'apical canal constriction' (apical constriction) were determined for each sp for both devices. The data were statistically analysed by a chi-square test. RESULTS: Precise determination of the target interval 'minor foramen-major foramen' was successful with Raypex 4 in 94.8% (sp 1), 90.7% (sp 0.5) and 72.5% (sp 0) of cases and with Justy II in 59.6% (sp 1), 92.2% (sp 0.5) and 68% (sp 0) of cases. No measurement carried out by Raypex 4 and by Justy II on sp 1 was beyond the major apical foramen. However, on sp 0.5, there were eight measurements for Raypex 4 and four measurements for Justy II beyond the major apical foramen. Overinstrumentation was also recorded for sp 0 in 49 specimens (Raypex 4) and 59 specimens (JustyII). The major apical constriction was met exactly by Raypex 4 in 50.7% (sp 1), 14% (sp 0.5) and 5.2% (sp 0) of cases and by Justy II in 32.1% (sp 1), 23.8% (sp 0.5) and 4.1% (sp 0) of cases. The differences between the determination made with the sp suggested by the manufacturers for Raypex 4 (sp 1) and Justy II (sp 0.5) were not significant (P > 0.05) for the target interval 'minor foramen-major foramen' and significant (P 0.05). CONCLUSIONS: Porphyromonas endodontalis enhances t-PA production in human pulp cells, and the signal transduction pathways p38 and MEK are involved in the inhibition of t-PA. PMID- 16343112 TI - Ex vivo study of the adhesion of an epoxy-based sealer to human dentine submitted to irradiation with Er : YAG and Nd : YAG lasers. AB - AIM: To evaluate the adhesion of an epoxy-based sealer to human dentine submitted to irradiation with Er : YAG or Nd : YAG laser at various parameters. METHODOLOGY: Ninety maxillary canine teeth were sectioned transversely at the cemento-enamel junction and at the root tip to leave an 8-mm-long cylinder. The tooth specimen was centred in a metallic ring (16 mm diameter and 8 mm height) and embedded in acrylic resin. The root canals were prepared using a low-speed handpiece and a conical diamond bur, which was attached to a paralleling device. This bur was lowered to a depth previously determined by a silicone stop. Specimens were divided into nine groups: group I, dentine was treated with 2 mL of 17% EDTAC for 5 min. Groups II-V were irradiated with Er : YAG laser at the following parameters: group II - 8 Hz and 200 mJ input (120 mJ output); group III - 8 Hz and 400 mJ input (240 mJ output); group IV - 16 Hz and 200 mJ input (120 mJ output); group V - 16 Hz and 400 mJ input (240 mJ output). Groups VI-IX were irradiated with Nd : YAG laser at the following parameters: group VI - 10 Hz and 1 W input (0.4 W output); group VII - 10 Hz and 2 W input (0.8 W output); group VIII - 15 Hz and 1 W input (0.4 W output); group IX - 15 Hz and 2 W input (0.8 W output). The root canals were filled with an epoxy-based root canal sealer and submitted to a push-out test. RESULTS: Statistical analysis showed significant differences (P < 0.01) between Er : YAG and Nd : YAG laser treatments at the higher frequencies compared with 17% EDTAC. Greater adhesion values were obtained for groups IV and V (Er : YAG laser) and groups VIII and IX (Nd : YAG laser), which were statistically different from groups II and III (Er : YAG laser) and groups VI and VII (Nd : YAG laser). Treatment with only 17% EDTAC had the lowest adhesion values. CONCLUSIONS: An increase in frequency, independent of power settings, of the lasers used in this study increased adhesion of an epoxy-based root canal sealer. PMID- 16343113 TI - Apical extrusion of intracanal bacteria following use of two engine-driven instrumentation techniques. AB - AIM: To evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the two engine-driven techniques utilizing nickel-titanium instruments (ProTaper and System GT). METHODOLOGY: Forty extracted single-rooted human mandibular premolar teeth were used. Access cavities were prepared and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into two experimental groups of 15 teeth each and one control group of 10 teeth. Group 1, ProTaper group: the root canals were instrumented using ProTaper instruments. Group 2, System GT group: the root canals were instrumented using System GT instruments. Group 3, control group: no instrumentation was attempted. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units. The data obtained were analysed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. RESULTS: There was no significant difference as to the number of extruded bacteria between the ProTaper and System GT engine driven systems (P > 0.05). CONCLUSIONS: Both engine-driven nickel-titanium systems extruded bacteria through the apical foramen. PMID- 16343115 TI - Heat shock induces the synthesis of the inflammatory mediator leukotriene B4 in human pulp cells. AB - AIM: To measure the synthesis of leukotriene B4 (LTB4) in cultures of human dental pulp cells induced by heat shock. METHODOLOGY: Primary pulp cells (PC) and dental pulp stem cells (DPSC) were cultivated under appropriate conditions. For the characterization of PC the expression of dentine sialophosphoprotein (DSPP) was evaluated by reverse transcription-polymerase chain reaction. Thermal stimulation of cell cultures was performed at temperatures of 37, 38, 39, 40, 42 and 45 degrees C for stimulation times of 5 and 30 s. LTB4 was quantified by reversed-phase high-performance chromatography and differences between the LTB4 concentrations of controls and heat stimulated cells were analysed with Friedman analysis of variances by ranks and multiple comparisons (P < 0.05). RESULTS: Both cell cultures expressed DSPP under the conditions of the present experiment. The analysis revealed significantly enhanced LTB4 synthesis following thermal stimulations at 38, 39, 40, 42 and 45 degrees C compared with unstimulated controls for both PC and DPSC. CONCLUSION: The present study demonstrated the capability of pulp cells to synthesize the arachidonic acid mediator LTB4 in response to heat shock. LTB4 has the capacity to induce inflammatory reactions and to sensitise afferent nociceptive nerve endings. LTB4 synthesis is induced by minor temperature changes, which are relevant for various clinical situations. PMID- 16343114 TI - The role of preoperative prophylactic antibiotic administration in periapical endodontic surgery: a randomized, prospective double-blind placebo-controlled study. AB - AIM: To determine the value of clindamycin prophylaxis in the prevention of postoperative wound infections in patients undergoing endodontic surgery. METHODOLOGY: This study included 256 patients undergoing endodontic surgery in a prospective double-blind placebo-controlled trial comparing oral administration of an oral placebo versus a preoperative 600 mg dose of clindamycin. After randomization the study medication was administered orally 1 h before surgery in a double-blind fashion. For a period of 4 weeks the postoperative course was observed according to clinical parameters of infection. Primary end-point was infection at the surgical site. RESULTS: The mean age of the study population was 44.4 years (SD 11.4, range 18-82 years) with a sex distribution of 147 females (47.4%) and 109 males (42.6%). Mean age of the patients in the clindamycin group was 44.7 years (SD 12.0), and the mean age in the placebo group was 44.1 years (SD 10.8) (P = 0.49). In the clindamycin group, the mean duration of surgery was 32.3 min (SD 8.8) and in the placebo group the mean duration of surgery was 32.5 min (SD 8.4) (P = 0.89). Two infections [1.6%; 95 confidence interval (CI): 0.48 4.72] were identified in the clindamycin group and four (3.2%; 95 CI: 0.42-1.33) in the placebo group (P = 0.448). CONCLUSIONS: No statistically significant difference was found between clindamycin prophylaxis and placebo with regard to the prevention of postoperative infection in endodontic surgical procedures. PMID- 16343116 TI - Calcium hydroxide dressings using different preparation and application modes: density and dissolution by simulated tissue pressure. AB - AIM: To study the effect of different apical shapes in prepared simulated root canals on the application of a commercially prepared calcium hydroxide paste by a syringe or lentulo spiral. METHODOLOGY: Three different types of root canal preparation were performed in 90 simulated canals: group A to an apical size 20 and a 0.10 taper using hand and rotary instruments, group B to an apical size 30 and a 0.08 taper using GT rotary instruments and group C to an apical size 40 and a 0.04 taper using ProFile 0.04 instruments. The insertion of calcium hydroxide [Ca(OH)2] paste was accomplished using either a lentulo spiral or a syringe. After 1 week of simulated fluid pressure applied to the apical end of the canal using physiological saline solution, the solution was evaluated for released Ca(OH)2. The specimens were weighed initially, after preparation, after insertion of Ca(OH)2 paste, after temporization with Cavit and after 1 week of simulated fluid pressure. Digital radiographs of the filled canals were taken and canal areas in mm2, gray values of the Ca(OH)2 dressings, total area of voids in mm2, as well as location of voids in the apical, middle or coronal thirds of the root canals were measured. Analyses of variance, with Scheffe's post-hoc tests, as well as chi-square tests were performed. RESULTS: Canals in group C had significantly fewer (P < 0.01) radiographic voids than canals in groups A and B. Using a lentulo spiral resulted in significantly (P < 0.05) fewer voids compared with the injection technique. More voids were detected coronally compared with middle and apical root canal thirds (P < 0.05). CONCLUSIONS: Canal shape and method of application had an impact on the amount and radiodensity of calcium hydroxide dressings in simulated root canals. Canals prepared to an apical size 40 and a taper of 0.04 had the least number of voids; Ca(OH)2 was placed with significantly fewer voids using a lentulo spiral compared with the injection technique. PMID- 16343117 TI - Effect of mineral trioxide aggregate on cytokine production by peritoneal macrophages. AB - AIM: To test the effect of two commercial brands of grey mineral trioxide aggregate (ProRoot and MTA-Angelus) on cytokine production by M1 and M2 inflammatory macrophages. METHODOLOGY: M1 (from C57BL/6 mice) and M2 peritoneal inflammatory macrophages (from C57BL/6 IL12p40-/- mice) were obtained and cultured in vitro in the presence of MTA. The cellular viability and the production of tumour necrosis factor-alpha, interleukin (IL)-12 and IL-10 in response to stimulation with interferon-gamma and Fusobacterium nucleatum or Peptostreptococcus anaerobius were evaluated. Data were analysed by Mann-Whitney, Kruskal-Wallis and anova tests. RESULTS: The cements did not interfere with cellular viability or with cytokine production by either type of macrophage. However, M2 macrophages produced higher levels of IL-10 when stimulated with F. nucleatum than M1 macrophages (P < 0.05). CONCLUSIONS: The brands of MTA evaluated did not interfere in the cytokine response by M1 or M2 macrophages to the two bacteria tested. However, a difference in cytokine production between the two types of macrophages was found. PMID- 16343118 TI - Three-dimensional, non-destructive visualization of vertical root fractures using flat panel volume detector computer tomography: an ex vivo in vitro case report. AB - AIM: To detect and to visualize radiographically vertical root fractures in extracted teeth with a prototype of a novel, high resolution, three-dimensional flat panel volume detector computer tomograph (FD-VCT) system. SUMMARY: Five teeth with root fillings and clinical symptoms such as fistulas and isolated periodontal pockets of 8 mm or more were extracted after dental radiography indicating lateral or periapical lesions. Vertical root fractures or cracks were suspected because of the symptoms and clinical findings were evident after extraction in all cases but fracture lines were not visible on routine dental radiographs acquired before extraction. The extracted teeth were explored with a prototype of a FD-VCT. Using the FD-VCT, in all cases vertical root fractures or crack lines could be detected clearly in different views, depiction-modes and cross-sections at a spatial resolution of 140 microm. The evaluation of the fracture lines and teeth could be performed in three-dimensional views. The FD VCT findings were confirmed by detailed inspection of the extracted teeth. KEY LEARNING POINTS: The FD-VCT is an innovative diagnostic tool for non-destructive, three-dimensional evaluation of extracted teeth in pre-clinical and experimental studies. The FD-VCT allows precise visualization and evaluation of vertical root fractures or cracks in extracted teeth. Clinical application of the system may be possible if technical modifications reduce the exposure dose: the high resolution detector systems of the FD-VCT should be combined with radiation systems that focus the radiation to the area of interest. PMID- 16343123 TI - Incidence of corneal pigmented arc and factors associated with its appearance in orthokeratology. AB - PURPOSE: To determine the incidence of the corneal pigmented arc in orthokeratology (ortho-k) lens wearers over 12 months of lens wear and the factors associated with its appearance. METHOD: Thirty-five ortho-k subjects were recruited; refractive and corneal changes after lens wear (single-lens protocol) were monitored over 12 months. The incidence of the pigmented arc after 3, 6 and 12 months of lens wear was determined. RESULTS: The incidence of corneal pigmented arc was 17% (27%), 49% (49%) and 90% (93%) after 3, 6 and 12 months lens wear respectively in the left and right eyes. For subjects with arcs observed in the left eye within the first 6 months of lens wear, the mean +/- S.D. period of lens wear before initial detection of the arc was 14 +/- 7.4 weeks, and no correlation was found between this factor and the baseline spherical and cylindrical refractive errors (i.e. refractive sphere and cylinder, respectively), spherical equivalent refractive error (SERE), the target myopia reduction, the amount of refractive sphere (or SERE) reduction and changes in central and peripheral corneal curvatures after 6 months of lens wear. Baseline refractive sphere, baseline SERE, target, amount of myopia reduction, and change in central corneal curvature were significantly larger (p < 0.05) in those subjects with pigmented arcs after about 6 months of lens wear. The intensity of the observed pigmented arcs after about 6 months of lens wear was significantly related to the time when it was first observed (p = 0.003). Significant correlation was also found between the intensity of the arcs and the following parameters: baseline refractive sphere and SERE, target, change in central corneal curvature, and amount of myopia reduction (p < 0.006). After about 12 months of lens wear, the intensity of observed arcs was significantly related to the baseline refractive sphere, SERE and the target (p < 0.006). CONCLUSION: The incidence of ortho-k-associated pigmented arc increases from 17% after 3 months of lens wear to over 90% after 12 months of lens wear. The intensity of the arc is related to the time when the arc first appeared. Both the incidence and the intensity of the arc are related to the period of lens wear, baseline refractive sphere, SERE and the target. PMID- 16343124 TI - Increase in central corneal thickness in dry and normal eyes with application of artificial tears: a new diagnostic and follow-up criterion for dry eye. AB - AIMS: To investigate the effect of artificial tear application on central corneal thickness (CCT) in dry and normal eyes. METHODS: Forty eyes of 40 patients with symptomatic dry eyes and 20 eyes of 20 controls were included in the study. A combination artificial tear therapy was given to both groups for 1 week. The CCT was measured with ultrasonic pachymetry before and after the therapy. In addition, at the beginning of the study, 11 dry eye patients and eight control patients were evaluated separately as a subgroup: they received artificial tears every 10 min for an hour to determine whether this might constitute a diagnostic criterion. Results were analysed with a Wilcoxon test, accepting p < 0.01 as significant. RESULTS: The mean CCT in the dry eye group before and after 1-week therapy was 531.5 +/- 16.5 and 559.6 +/- 26.1 microm, respectively (p < 0.0001); thus yielding an average CCT increase of 28.1 +/- 9.4 microm (=5.30%: 1.65% minimum; 7.03% maximum) from baseline. The mean CCT in the control group before and after 1-week therapy was 549.2 +/- 37.8 and 552.3 +/- 39.8 microm, respectively (p = 0.001); thus, yielding an average CCT increase of 3.1 +/- 1.9 microm (=0.56%: 0.00% minimum; 1.18% maximum) from baseline. When the subgroup was evaluated separately, the average increase from baseline in 11 dry eye patients was 10.4 +/- 4.0 microm (=1.89%: 0.60% minimum; 4.26% maximum) after 1 h of artificial tear application, while it was 2.9 +/- 2.0 microm (=0.51%: 0.00% minimum; 0.58% maximum) in eight control patients (p < 0.0001). CONCLUSION: The CCT in dry eyes increases rapidly and significantly compared to normal eyes after application of artificial tears. It is suggested that this increase could be used as a criterion in the diagnosis and follow up of dry eyes, and that this increase in thickness should be considered in intra-ocular pressure measurements as well as in refractive surgical procedures. PMID- 16343125 TI - Evaluation of retinal function using the Dynamic Focal Cone ERG. AB - The development of effective means of assessing visual function in retinal disease holds the key to improved understanding of pathogenesis, and better monitoring of treatment outcomes. In diseases such as age-related macular degeneration, in which the primary locus of dysfunction is the outer retina, tests which provide a direct measure of the functional integrity of the photoreceptor/retinal pigment epithelium (RPE) complex are of great importance. Recovery of retinal function following adaptation to a bright light requires the healthy function of photoreceptors, RPE, Bruch's membrane and choroidal circulation, making an assessment of this recovery a potentially useful clinical tool. However, current techniques are either subjective in nature, or are influenced by post-retinal processing of visual information. This report describes a novel technique, the 'Dynamic Focal Cone Electro-retinogram (ERG)', which allows direct, objective assessment of the recovery of macular function following photopigment bleach. A series of 41 Hz ERGs was recorded, and ERG amplitude was plotted as a function of time following cessation of the bleach. Normative data was collected from 10 healthy subjects. For all subjects, there was no measurable ERG immediately after the bleach, but the amplitude had returned to a pre-bleach level within 4 min. The amplitude recovery data were adequately described both by an exponential recovery function and by a model based on a rate-limited recovery process. We conclude that this technique provides a clinically applicable, objective measure of outer retinal recovery. PMID- 16343126 TI - Chromatic and achromatic transient VEPs in adults with Down syndrome. AB - Oculo-visual abnormalities such as strabismus and high refractive error are common in people with Down syndrome, and account in large part for reduced visual function in this group. In the absence of such abnormalities, however, some spatial vision deficits persist, probably reflecting abnormal function of the neural visual pathway in this population. In addition, colour vision abnormalities are reportedly common in subjects with Down syndrome. We recorded transient visual evoked potentials in response to black-white and chromatic stimuli, in seven subjects with Down syndrome and 33 controls, to investigate function of the visual pathways underpinning spatial and chromatic visual function in Down syndrome. Our findings indicate, in agreement with previous studies, that retino-striate achromatic and chromatic processing in Down syndrome are abnormal. We find, however, that abnormal retino-striate processing of chromatic signals in this group may not give rise to colour vision deficits detected by the Colour Vision Test Made Easy or the City University test. PMID- 16343127 TI - In focus: the use of bifocal spectacles with children with Down's syndrome. AB - PURPOSE: Over 75% of children with Down's syndrome fail to accommodate accurately on near targets. This deficit must result in optically blurred images for near work. This present study set out to evaluate the controlled use of bifocal spectacles as an aid to near focusing. Although sometimes used clinically, no systematic studies of bifocals for children with Down's syndrome have been reported. METHODS: This was a comparative non-randomised interventional study. Thirty-four children with Down's syndrome of primary school age (5-11 years) took part, assigned in equal numbers to form two matched groups. All children received a full optometric assessment prior to entering the study. The treatment group was prescribed bifocal spectacles with a +2.50 addition, and the control group provided with single vision lenses to correct any clinically significant refractive error. Three follow-up visits were made over a 5-month period after spectacle provision. RESULTS: The treatment group showed consistently more accurate accommodation than the control group both through the bifocal segment, and, unexpectedly, through the distance part of the lens (p < 0.05). Compliance with new spectacles was high in both groups (>82% fully compliant). CONCLUSIONS: Bifocals confer benefit to children with Down's syndrome who under-accommodate, both directly (better focusing through the bifocal) and indirectly (by encouraging improved accommodation through the distance part of the lens). Based on the results of this study, eye examinations of children with Down's syndrome should routinely include a measure of accommodation at near, and bifocal spectacles should be considered for those who show under-accommodation. PMID- 16343128 TI - The management of systemic hypertension in optometric practice. AB - AIM: Systemic hypertension is a silent killer that may have very few warning signs. This study examines detection and management of patients with cardiovascular disease (CVD) by optometrists in the UK. METHOD: A survey was sent out to 1402 optometrists who practice in the UK to investigate the nature and knowledge of detection of systemic hypertension. RESULTS: The survey response rate was 37%. Optometrists were neutral to slightly positive towards the use of blood pressure (BP) monitors, but not on a routine basis. The most frequently asked question during history and symptoms concerned the patient's prescribed medication for systemic hypertension, and the least was that of specifying the last BP measurement. The additional test most widely used in the examination of the systemic hypertensive patient was direct ophthalmoscopy with red-free filter. Over one-tenth of optometric practices had a BP monitor, with automated devices being the most popular. Patients most likely to have their BP measured were suspect systemic hypertensives. Retinal haemorrhages are the most important finding in influencing an optometrist's referral criteria for suspected systemic hypertension. CONCLUSION: Although optometrists monitor the retina for signs of damage from systemic hypertension, further education in this important condition is warranted. PMID- 16343129 TI - Visual function assessment and metamorphopsia after macular hole surgery. AB - The purpose of this study was to develop a new resolution acuity measure for patients after macular hole surgery. Fifty eyes of 44 patients who had undergone successful treatment were tested. Visual acuity was measured with the Snellen, Early Treatment of Diabetic Retinopathy Study and Landolt-C charts. A Line Resolution Test was performed as part of their follow-up exam where a vertical line was presented. Participants were categorized by their perception of the line as solid, bent or broken. The line could be distorted into a sine-wave pattern in order to determine the participants' detection threshold for the distortion. Chart acuities did not differ among the three groups, as categorized by their line perception. Only the distortion measure was sensitive enough to differentiate the solid- from the broken-line group. The distortion measure assesses resolution power of the macula in smaller increments than acuity charts. This hyperacuity approach is more appropriate in the assessment of functional outcome after microsurgery. PMID- 16343131 TI - A theoretical study of intraocular lens tilt and decentration on perceptual image quality. AB - Maps of perceptual image quality were generated for a range of intraocular (IOL) tilts and decentrations, using a perceptually based metric i.e. the square integral (SQRI) metric. Maps were generated for a well-corrected eye, and for hyperopic (-0.1 mm) and myopic (+0.1 mm) image planes (for 2 and 4 mm pupils). These perceptually based results were compared with a standard optical measure (i.e. the equivalent SQRI for given spectacle power added at the cornea). The image quality changes in maps were small, but detectable (exceeding the perceptual just noticeable difference (jnd) or 1 jnd threshold). Clockwise (negative) tilt combined with decenter fell below the 'contrast reserve' modulation transfer functions (MTFs) for 0.25 and 0.5 D defocus, 4 mm pupil. These errors were larger than those found previously, probably because of the shape-factor of the IOL used. Pupil size contributed the largest drop in SQRI value, followed by clockwise tilt combined with decenter. Anticlockwise tilt with decenter produced image quality improvements for the hyperopic eye. Spot diagrams confirmed defocus and the balance of coma-like aberration and astigmatism as causes of the changes in SQRI values. The SQRI maps conveniently summarize imaging, and may be a useful tool in assessing image quality in eye modelling applications. PMID- 16343130 TI - Recognition performance and perceived quality of video enhanced for the visually impaired. AB - Subjects with central field loss (CFL) individually selected enhancement parameters to improve visibility of static video images. The effect of enhancement on performance and on perceived quality of motion video was assessed. Performance, e.g. recognition of visual details, was assessed by having subjects answer questions regarding visual information contained in the video motion segments that were enhanced using the individually-selected parameters. Enhancement did not improve subject performance on questions about video content. This result might be due to a ceiling effect limitation of the performance assessment method. In a second procedure, subjects' continuous perceptions of quality (using an adjective-based rating scale) were made while the enhancement parameters were abruptly switched among multiple values; these included the individually-selected enhancements, as well as unenhanced, over-enhanced, and degraded segments. The results indicate that adaptive enhancement (individually tuned using a static image) adds significantly to perceived image quality when viewing motion video. Subjects who selected stronger contrast enhancement also perceived the enhancement to provide a larger benefit in image quality. PMID- 16343132 TI - Impact of decentration of astigmatic intra-ocular lenses on the residual refraction after cataract surgery. AB - PURPOSE: The purpose of this study is to assess the impact of decentration of astigmatic intra-ocular lenses on the residual refraction after cataract surgery, using a computing scheme with 5 x 5 system matrices. METHODS: Based on the definition of an optical system in the paraxial Gaussian space containing astigmatic surfaces without restrictions to coaxiality, we derived a method (using 5 x 5 refraction and translation matrices) for calculating the residual refraction and the compensating prism in the spectacle plane after decentred implantation of thin and thick astigmatic intra-ocular lenses. The 'optical system eye' may contain astigmatic refractive surfaces with their axes at random. RESULTS: The capabilities of this computing scheme are demonstrated with two examples. In example 1 we calculate the residual refraction of a decentred 'thin astigmatic lens' for compensation of corneal astigmatism to achieve a spherical target refraction. In example 2 we compute the residual refraction after implantation of a 'thick astigmatic lens', where the spherical and cylindrical power as well as the implantation axis of the lens do not fully match the pre operative recommendations and the lens is decentred relative to the optical axis. For both examples, we derive the residual prismatic effect in the spectacle plane and the lateral displacement of a ray exiting the spectacle correction when starting coaxially at the retina. CONCLUSIONS: We have presented an en bloc matrix-based strategy for the calculation of the residual spherocylindrical refraction at the spectacle plane after implantation of a decentred thin or thick astigmatic intra-ocular lens without restrictions to coaxiality. The resulting system matrix is written as a product of 5 x 5 refraction and translation matrices. PMID- 16343133 TI - Compensation of aniseikonia in astigmatic pseudophakic eyes. AB - PURPOSE: A recently published manuscript addressed the problem of compensating for aniseikonia between pseudophakic astigmatic eyes using a least-squares calculation scheme. The purpose of this paper is to revisit this topic with the specific aim of providing explicit formulae for the determination of the intra ocular lens required to produce a specified transverse image size at the plane of the retina and the characteristics of the contact or spectacle lens required to realize some desired refractive outcome. METHODS: The 4 x 4 ray transference is central to the development of all formulae presented in this paper. Specifically, the formula for the determination of the power of the intra-ocular lens required to achieve some transverse image size at the retina is derived directly from the disjugacy of the pseudophakic eye. RESULTS: The formula is applicable to both stigmatic and astigmatic systems without restriction. A detailed numerical example for an unusual eye is provided. CONCLUSION: A formula for the determination of the intra-ocular lens required to produce any given transverse image size at the retina is derived. This approach does not require the application of the Moore-Penrose pseudo-inverse and one is able to work rather with the properties of the optical system directly without further modification. PMID- 16343134 TI - Accuracy of accommodation in heterophoric patients: testing an interaction model in a large clinical sample. AB - A model of the cross-link interactions between accommodation and convergence predicted that heterophoria can induce large accommodation errors (Schor, Ophthalmic Physiol. Opt. 1999;19:134-150). In 99 consecutive patients with intermittent tropia or decompensated phoria, we tested these interactions by comparing their accommodative responses to a 2.50-D target under binocular fused conditions (BFC) and monocular occluded conditions (MOC). The accommodative response in BFC frequently differed from that in MOC. The magnitude of the accommodative errors in BFC, ranging from an accommodative lag of 1.80 D (in an esophoric patient) to an accommodative lead of 1.56 D (in an exophoric patient), was correlated with distance heterophoria and uncorrected refractive errors. These results indicate that heterophoria affects the accuracy of accommodation to various degrees, as the model predicted, and that an accommodative error larger than the depth of focus of the eye occurs in exchange for binocular single vision in some heterophoric patients. PMID- 16343135 TI - Amblyopia prevalence and cigarette smoking by women. AB - The relationship between cigarette smoking by young women and amblyopia in children was investigated in 13 population groups. Two searches were conducted on Medline using the following keywords for the first search: amblyopia, prevalence or incidence, and the names of specific countries. The keywords for the second search were smoking, cigarettes, women and the names of the same specific countries. Relevant articles were reviewed. A positive relationship between the rate of smoking among women and the prevalence of amblyopia in school-aged children and military recruits was found. The Pearson correlation coefficient was 0.73. The occurrence of decreased visual acuity attributed to amblyopia in disparate population groups is directly related to rates of cigarette smoking by young women. PMID- 16343138 TI - A new approach for perineural injection of the lateral palmar nerve in the horse. AB - OBJECTIVE: To evaluate the accuracy of a new technique for perineural injection of the lateral palmar nerve and to determine frequency of inadvertent injection into the carpal synovial sheath with this technique. STUDY DESIGN: Prospective experimental study. ANIMALS: Thirty equine cadaver forelimbs. METHODS: Each of 3 clinicians injected 0.5 mL of a 1% aqueous solution of new methylene blue as a marker at the medial aspect of the accessory carpal bone of 10 limbs. Immediately after each injection, the lateral palmar nerve was identified by dissection of and inspected for proximity of dye, and the carpal synovial sheath was inspected for the presence of dye. RESULTS: New methylene blue solution was observed to surround the nerve (29 limbs) or to lie within 2 mm of it (1 limb). Dye was not found in the carpal synovial sheath of any specimen. CONCLUSIONS: Using this technique, perineural injection of the lateral palmar nerve can be consistently achieved, and the carpal synovial sheath is unlikely to be penetrated by the needle during the procedure. CLINICAL RELEVANCE: The technique described provides an accurate and simple method for perineural injection of the lateral palmar nerve proximal to the origin of its deep branch. This technique can be used to anesthetize the lateral palmar nerve for diagnosis of pain originating in the palmaroproximal aspect of the metacarpus without risk of inadvertently desensitizing structures within the carpal synovial sheath. PMID- 16343139 TI - Upper airway dysfunction associated with collapse of the apex of the corniculate process of the left arytenoid cartilage during exercise in 15 horses. AB - OBJECTIVE: To report dynamic collapse of the apex of the left corniculate process under the right corniculate process into the airway at the dorsal apposition of the paired arytenoid cartilages during exercise as a cause of upper airway dysfunction in horses. DESIGN: Retrospective study. ANIMALS: Fifteen horses with a history of poor performance and/or upper respiratory tract noise during exercise. METHODS: Video recordings of all horses referred for upper airway evaluation using high-speed treadmill videoendoscopy (HSTV) between January 1998 and December 2003 were reviewed. Records of horses that developed dynamic collapse of the apex of the left corniculate process into the airway were included. Clinical history, age, gender, breed, and use of the horse were retrieved. RESULTS: Of 309 horses referred for examination for poor performance and/or upper respiratory tract noise during exercise, 15 (4.9%) had collapse of the apex of the left corniculate process under the right and into the airway at the dorsal apposition between the paired arytenoid cartilages during HSTV. There were 3 females and 13 males, aged from 2 to 5 years. Five horses had previous surgery for left recurrent laryngeal neuropathy (RLN): 2 had nerve muscle pedicle graft and 3 had laryngeal prosthesis. During HSTV, all 15 horses had progressive collapse of the apex of the left corniculate process under the right at the dorsal apposition of the 2 arytenoid cartilages, and into the dorsal aspect of the rima glottidis. Review of video recordings revealed that collapse of the apex of the corniculate process was followed by progressive collapse of the left aryepiglottic fold and left vocal fold. The ventral aspect of the left corniculate cartilage maintained abduction in all horses. Two horses also had progressive collapse of the right vocal fold, 1 had rostral displacement of the palatopharyngeal arch, and another had dorsal displacement of the soft palate. CONCLUSIONS: Dynamic collapse of the apex of the left corniculate process of the arytenoid cartilage under the right is an uncommon cause of upper airway dysfunction in horses and the pathogenesis is unclear. We speculate that the left arytenoideus transversus muscle is unable to support the dorsal apposition between the arytenoid cartilages. This loss of support allows the elastic cartilage of the left corniculate process to collapse under the right and into the airway, as inspiratory pressure increases during exercise. This condition may be associated with an unusually advanced neuropathy of the adductor components of the left recurrent laryngeal nerve and may be an unusual manifestation of RLN; however, this is speculative and further investigation is required to determine its cause. CLINICAL RELEVANCE: Dynamic collapse of the apex of the left corniculate process and into the airway at the dorsal apposition between the paired arytenoid cartilages can only be diagnosed during HSTV. It is an uncommon cause of upper airway dysfunction but may affect the athletic potential of racing Thoroughbreds and Standardbreds. PMID- 16343140 TI - Evaluation of a canine cranial cruciate ligament repair system for use in equine laryngoplasty. AB - OBJECTIVE: To report the use of a nylon suture system (Canine Cranial Cruciate Ligament Repair System; Securos Inc Veterinary Orthopedics) as a prosthesis for equine laryngoplasty. STUDY DESIGN: Experimental and prospective clinical study. ANIMALS: Cadaver specimens (n = 5) and 7 horses with left laryngeal hemiplegia. METHODS: A commercially available monofilament nylon suture system was implanted as a laryngeal prosthesis. Arytenoid cartilage abduction was achieved with a tensioning device applied to the suture prosthesis during transnasal endoscopic observation. Suture fixation was achieved with crimping clamps and a crimping device. RESULTS: The nylon suture system was suitable as a laryngeal prosthesis for arytenoid cartilage abduction. The ratchet mechanism of the tensioning device facilitated abduction of the arytenoid cartilage and suture fixation was achieved by the crimped clamp without any loss of tension. Postoperatively, there was a slight loss of tension in 4 horses and complete loss of tension in 1 horse because of cartilage failure. After convalescence, none of the horses had abnormal respiratory noise, exercise intolerance or cough. CONCLUSIONS: A nylon suture system designed for canine cranial cruciate ligament repair was used successfully as a laryngeal prosthesis and facilitated control of the degree of arytenoid cartilage abduction during laryngoplasty. CLINICAL RELEVANCE: For improved control of the degree of arytenoid cartilage abduction during laryngoplasty, use of a nylon suture system with metal crimps should be considered. PMID- 16343141 TI - Investigation of the immediate analgesic effects of extracorporeal shock wave therapy for treatment of navicular disease in horses. AB - OBJECTIVES: To measure the acute analgesic properties of extracorporeal shock wave therapy (ESWT) in horses with navicular disease using objective ground reaction forces (GRF). STUDY DESIGN: Prospective study. ANIMALS: Nine client owned horses with unilateral forelimb lameness caused by navicular disease. METHODS: Horses were trotted at 3 m/s over a force plate to record pre-treatment GRF of the lame and contra-lateral forelimbs. After ESWT of the lame limb, applied with a radial unit to each of the frog and heel bulb areas with 1500 pulses at 4 bar and 10 Hz, GRF measurements were repeated at 15 minutes and 24 hour intervals for 7 days. From the 5 trials recorded at each time, peak vertical force (PVF) was calculated and averaged. Data were compared between limbs and over time using a 2-way repeated-measures ANOVA. RESULTS: Mean (+/-SD) PVF was significantly different between lame (day 1; 4339+/-626 N) and contra-lateral (day 1; 5236+/-515 N) limbs pre-treatment (P<.05). After ESWT treatment, this difference remained unchanged at 15 minutes and from days 1 to 7. To confirm navicular disease, palmar digital analgesia significantly reduced the lameness (lame 5144+/-430 N; contralateral 5082+/-586 N; P>.05). CONCLUSIONS: Single ESWT treatment applied in this manner does not influence lameness in horses with navicular disease. CLINICAL RELEVANCE: Although many equine regulatory commissions currently ban the use of ESWT before competition, our results suggest that such regulations may be unfounded. Further research is required to determine the long-term analgesic properties of this frequently used treatment modality in equine athletes. PMID- 16343142 TI - Use of the Anderson Sling suspension system for recovery of horses from general anesthesia. AB - OBJECTIVE: To describe a sling recovery system (Anderson Sling) for horses and to evaluate outcome of high-risk horses recovered from general anesthesia by a sling. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Horses (n=24) recovered from general anesthesia. METHODS: Complete medical and anesthetic records (1996-2003) for horses recovered from general anesthesia using the Anderson Sling system were evaluated retrospectively. Information retrieved included anesthetic protocol, surgical procedure, recovery protocol, recovery time, and quality of the recovery. Horses were recovered from anesthesia supported by the Anderson Sling in a standing position within a traditional padded equine recovery stall. RESULTS: Twenty-four horses had 32 assisted recoveries; 31 events were successful. No complications associated with the sling or recovery system protocol occurred. One horse was intolerant of the sling's support and was reanesthetized and recovered successfully using head and tail ropes. CONCLUSION: The Anderson Sling recovery system is an effective and safe way to recover horses that are at increased risk for injury associated with adverse events during recovery from general anesthesia. CLINICAL RELEVANCE: The Anderson Sling system should be considered for assisted recovery of equine patients from general anesthesia. PMID- 16343143 TI - Ex vivo strength comparison of bioabsorbable tendon plates and bioabsorbable suture in a 3-loop pulley pattern for repair of transected flexor tendons from horse cadavers. AB - OBJECTIVE: To test the failure strength and energy of 2 bioabsorbable implants applied to transected deep digital flexor tendons (DDFT) from adult horses. STUDY DESIGN: Ex vivo biomechanical experiment. SAMPLE POPULATION: Twelve pairs of deep digital flexor tendons harvested from the forelimbs of fresh equine cadavers. METHODS: Poly-L-lactic acid tendon plates were custom manufactured for application to the cylindrical surface of an adult equine deep digital flexor tendon. Twelve pairs of DDFTs were transected 2 cm distal to the insertion of the distal check ligament of the deep digital flexor tendon. One tendon of each pair was randomly selected for repair with a biodegradable plate or a 3-loop pulley method. Size 2 polydioxanone suture was used in both repairs. Repairs were tested in tension to failure, with peak force (PF) and total energy (TE) at repair failure recorded in Newtons (N) and Joules (J), respectively. A paired t-test was used for statistical evaluation with a significant level set at P< or = .05. RESULTS: Mean+/-SD PF for failure of plated tendons (1507.08+/-184.34 N) was significantly greater than for sutured tendons (460.86+/-60.93 N). TE was also significantly greater for failure of plated tendons versus sutured tendons. CONCLUSIONS: Plate fixation of transected cadaver DDFTs appear to have superior immediate failure strength than 3-loop pulley repairs. CLINICAL RELEVANCE: Whereas in vivo testing is required, a bioabsorbable tendon plate may provide initial increased strength to support tendon healing and decrease external coaptation requirements. PMID- 16343144 TI - Measurement of articular cartilage stiffness of the femoropatellar, tarsocrural, and metatarsophalangeal joints in horses and comparison with biochemical data. AB - OBJECTIVE: To determine normal cartilage stiffness values in different weight bearing and non-weight-bearing areas of 3 different equine joints, and to evaluate the relationship between cartilage stiffness and glycosaminoglycan (GAG) and collagen content. STUDY DESIGN: Compressive stiffness of the articular cartilage was measured in 8 horse cadaver femoropatellar (FP), tarsocrural (TC), and metatarsophalangeal (MT) joints. Gross evaluation, collagen content, GAG content, and histologic appearance were assessed for each measurement location. ANIMALS: Eight equine cadavers (4 intact females, 4 castrated males; 7 Quarter Horse or Quarter Horse type, 1 Arabian; aged 4-12 years, weighing 400-550 kg). METHODS: The articular surfaces of 8 equine cadaver FP, TC, and MT joints were grossly evaluated for signs of articular cartilage pathology. Stiffness at preselected sites (FP joint-6 sites; TC joint-3 sites; MT joint-4 sites) was determined using an arthroscopic indentation instrument. Biochemical composition (collagen, GAG content) and histologic evaluation (modified Mankin score) were assessed for each measurement site. RESULTS: All cartilage from all sites evaluated was determined to be normal based on macroscopic and histologic assessments. No significant correlation between Mankin scores and cartilage stiffness values was observed. Site differences in cartilage stiffness were measured in all 3 joints (P<.001). GAG or collagen content had a significant positive correlation with stiffness values in 6 of 13 sites (P<.05, r>0.622, r2>0.387). CONCLUSION: Relative cartilage stiffness values measured in healthy equine joints are site dependent and can be measured using an indentation device intended for arthroscopic application. CLINICAL RELEVANCE: An indentation instrument provided an objective means of determining relative compressive stiffness of articular cartilage. Further research needs to be performed to confirm the site and joint differences observed in this study in clinically normal horses and to determine if the tester can be used clinically to predict articular cartilage pathology. PMID- 16343145 TI - An in vitro biomechanical comparison of a limited-contact dynamic compression plate fixation with a dynamic compression plate fixation of osteotomized equine third metacarpal bones. AB - OBJECTIVES: To compare the monotonic biomechanical properties and fatigue life of a broad, limited contact, dynamic compression plate (LC-DCP) fixation with a broad, dynamic compression plate (DCP) fixation to repair osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. ANIMAL POPULATION: Twelve pairs of adult equine cadaveric MC3 bones. METHODS: Twelve pairs of equine MC3 were divided into 3 test groups (4 pairs each) for (1) 4-point bending single cycle to failure testing, (2) 4-point bending cyclic fatigue testing, and (3) torsional single cycle to failure testing. An LC-DCP (8-hole, 4.5 mm) was applied to the dorsal surface of 1 randomly selected bone from each pair. One DCP (8-hole, 4.5 mm broad) was applied dorsally to the contralateral bone from each pair. All plates and screws were applied using standard AO/ASIF techniques to MC3 bones that had mid diaphyseal osteotomies. Mean test variable values for each method were compared using a paired t-test within each group. Significance was set at P<.05. RESULTS: The mean 4-point bending yield load, yield bending moment, composite rigidity, failure load, and failure bending moment of LC-DCP fixation were significantly greater (P<.01) than those of broad DCP fixation. Mean cycles to failure for 4 point bending was significantly (P<.001) greater for broad DCP fixation compared with broad LC-DCP fixation. Mean yield load, mean composite rigidity, and mean failure load in torsion was significantly (P<.02) greater for broad LC-DCP fixation compared with broad DCP fixation. CONCLUSION: Broad LC-DCP offers increased stability in static overload testing, however, it offers significantly less stability in cyclic fatigue testing. CLINICAL RELEVANCE: The clinical relevance of the cyclic fatigue data supports the conclusion that the broad DCP fixation is biomechanically superior to the broad LC-DCP fixation in osteotomized equine MC3 bones despite the results of the static overload testing. PMID- 16343146 TI - An in vitro evaluation of plate luting using osteotomized equine third metacarpal bones with a limited contact-dynamic compression plate. AB - OBJECTIVES: To evaluate the effects of plate luting on the biomechanical properties of a broad limited contact-dynamic compression plate (LC-DCP) fixation to repair osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by LC-DCP fixation, with 1 of the pair luted with polymethylmethacrylate (PMMA). ANIMAL POPULATION: Ten pairs of adult equine cadaveric MC3 bones. METHODS: Ten pairs of equine MC3 were divided into 2 test groups (5 pairs each) for (1) palmarodorsal 4-point bending single cycle to failure testing and (2) palmarodorsal 4-point bending cyclic fatigue testing. The LC-DCP (8 hole, 4.5 mm) was applied to the dorsal surface of each pair of MC3 bones. All plates and screws were applied using standard AO/ASIF techniques. All MC3 bones had mid-diaphyseal osteotomies. One of the matched pairs of LC-DCP-MC3 constructs were randomly chosen to be luted with PMMA. Mean test variable values for each method were compared using a paired t-test within each group; significance was set at P<.05. RESULTS: Mean palmarodorsal 4-point bending yield bending moment, failure bending moment of the LC-DCP fixation with luting was not significantly different (P>.05) than those of the LC-DCP fixation without luting. Mean cycles to failure for palmarodorsal 4-point bending was significantly (P<.0003) greater, with a 7.2-fold increase, for the LC-DCP fixation with luting compared with the LC-DCP fixation without luting. CONCLUSION: Luting the broad LC DCP with PMMA in the fixation osteotomized equine MC3 bones increases the fatigue life of cyclic loading for palmarodorsal 4-point bending under the in vitro conditions studied. CLINICAL RELEVANCE: The cyclic fatigue data supports the conclusion that luted broad LC-DCP fixation is biomechanically superior to the non-luted broad LC-DCP fixation in osteotomized equine MC3 bones. PMID- 16343147 TI - An in vitro biomechanical comparison of a prototype equine metacarpal dynamic compression plate fixation with double dynamic compression plate fixation of osteotomized equine third metacarpal bones. AB - OBJECTIVES: To compare the monotonic biomechanical properties of a prototype equine third metacarpal dynamic compression plate (EM-DCP) fixation with a double broad dynamic compression plate (DCP) fixation to repair osteotomized equine third metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. POPULATION: Twelve pairs of adult equine cadaveric MC3 bones. METHODS: Twelve pairs of equine MC3 were divided into 3 test groups (4 pairs each) for (1) 4-point bending single cycle to failure testing, (2) 4-point bending cyclic fatigue testing, and (3) torsional testing. The EM-DCP (10-hole, 4.5 mm) was applied to the dorsal surface of one randomly selected bone from each pair. Two DCPs, 1 dorsally (10-hole, 4.5 mm broad) and 1 laterally (9-hole, 4.5 mm broad) were applied to the contralateral bone from each pair. All plates and screws were applied using standard AO/ASIF techniques to MC3 bones that had mid diaphyseal osteotomies. Mean test variable values for each method were compared using a paired t-test within each group. Significance was set at P<.05. RESULTS: Mean 4-point bending yield load, yield bending moment, bending composite rigidity, failure load and failure bending moment of the EM-DCP fixation were significantly greater (P<.0001) than those of the double broad DCP fixation. Mean cycles to failure in 4-point bending of the EM-DCP fixation was significantly greater (P<.0008) than that of the double broad DCP fixation. Mean yield load, composite rigidity, and failure load in torsion of the EM-DCP fixation were significantly greater (P<.0035) than that of the double broad DCP fixation. CONCLUSION: The EM-DCP provides increased stability in both static overload testing and cyclic fatigue testing. CLINICAL RELEVANCE: Results of this in vitro study support the conclusion that the prototype EM-DCP fixation is biomechanically superior to the double broad DCP fixation for the stabilization of osteotomized equine MC3. PMID- 16343148 TI - Effects of topical nitroglycerine patches and ointment on digital venous plasma nitric oxide concentrations and digital blood flow in healthy conscious horses. AB - OBJECTIVE: To measure and compare palmar digital venous plasma nitric oxide (NO) concentrations and digital arterial blood flow after application of topical nitroglycerine (NTG). STUDY DESIGN: Experimental study. ANIMALS: Healthy adult horses (n=8). METHODS: Digital blood flow was measured by an ultrasonic Doppler flow probe surgically implanted around the medial palmar digital artery. Blood was collected from a catheter placed in the medial palmar digital vein for quantification of NO. NTG patches, NTG ointment or control patches were placed over the palmar digital vessels at the level of the fetlock. Two horses had an intra-arterial infusion of an NTG solution into the medial palmar digital artery in a pilot study. RESULTS: Digital arterial blood flow did not change significantly with application of the NTG patches, NTG ointment, or control patches. There were no statistically significant or biologically important changes in digital venous NO concentrations across time or between treated and control horses. In the pilot study, digital arterial blood flow and palmar digital venous NO concentrations increased with intra-arterial infusion of NTG. CONCLUSIONS: In clinically healthy horses, digital arterial blood flow and digital venous plasma NO concentrations did not change significantly with application of the NTG patches/ointment. These treatments are unlikely to have an effect on the digital vasculature of laminitic horses, however, further investigation is warranted. CLINICAL RELEVANCE: Although NTG patches have been used as a method of decreasing vasomotor tone and improving digital blood flow in horses with laminitis, this study provides evidence in healthy conscious horses that this treatment is not effective in altering digital blood flow. PMID- 16343149 TI - Synovial fluid and plasma concentrations of ceftiofur after regional intravenous perfusion in the horse. AB - OBJECTIVE: To determine radiocarpal (RC) joint synovial fluid and plasma ceftiofur concentrations after regional intravenous perfusion (RIP) and systemic intravenous (IV) administration. STUDY DESIGN: Experimental cross-over study. ANIMALS: Five normal adult horses. METHODS: One RC joint was randomly selected for RIP and the contralateral RC joint was sampled to determine intrasynovial ceftiofur concentrations after IV administration. Wash-out between IV and RIP was > or = 14 days. After surgical introduction of an intraarticular catheter, ceftiofur (2 g) was administered under general anesthesia either IV or by RIP after tourniquet application. Plasma and synovial fluid were collected over 24 hours. Samples were analyzed using high-performance liquid chromatography with ultraviolet detection and the results were statistically analyzed using a linear mixed effect model. RESULTS: Mean synovial fluid ceftiofur concentrations were consistently higher after RIP than after IV administration and were > 1 mug/mL (minimal inhibitory concentration [MIC] for common pathogens) for >24 hours. Mean synovial fluid peak concentration of ceftiofur after RIP and IV administration was 392.7+/-103.29 microg/mL at 0.5 hours postinjection (HPI) and 2.72+/-0.31 mug/mL at 1 HPI, respectively. Large variations in synovial fluid and plasma ceftiofur concentrations were observed between horses regardless of administration technique. RIP did not cause adverse effects. CONCLUSIONS: Under the present experimental conditions RIP with ceftiofur (2 g) induced significantly higher intraarticular antibiotic concentrations in the RC joint in comparison with IV administration. Moreover, after RIP, synovial fluid ceftiofur concentrations remain above the MIC for common pathogens (1 microg/mL) for > 24 hours. No adverse effects from the technique or the antibiotic were observed. CLINICAL RELEVANCE: RIP with high doses of ceftiofur may be a beneficial adjunctive therapy when treating equine synovial infections which are caused by cephalosporin susceptible microorganisms. PMID- 16343150 TI - Medullary plasma pharmacokinetics of vancomycin after intravenous and intraosseous perfusion of the proximal phalanx in horses. AB - OBJECTIVE: To study the pharmacokinetics of vancomycin in plasma obtained from the medullary sinusoids of the proximal phalanx (P1) after intravenous (IV) and intraosseous (IO) regional limb perfusion (RLP). STUDY DESIGN: Experimental study. ANIMALS: Twelve horses. METHODS: IV and IO RLP were performed in 2 groups (n = 6) of horses. Vancomycin hydrochloride (300 mg in 60 mL 0.9% NaCl) was randomly infused in 1 front limb, with the contralateral limb as control (60 mL 0.9% NaCl). A hole was drilled in the lateral cortex of P1, and blood samples from the medullary sinusoids of P1 were collected before infusion, and 15, 30, 45, 65, and 90 minutes after beginning infusion. Samples were centrifuged and plasma vancomycin concentrations determined. Vancomycin concentrations were compared over time and between routes using a 2-way repeated measures ANOVA. Pharmacokinetic variables were compared with a Kolmogorov-Smirnov test. Significance was set at P<.05. RESULTS: No vancomycin-induced clinical side effects such as lameness or swelling were observed. Both techniques, IV and IO, produced high vancomycin concentrations in the sinusoidal plasma of the P1, which remained above the minimum inhibitory concentration value for methicillin resistant Staphylococcus aureus (MRSA) during the 90 minutes study. Concentrations and pharmacokinetic variables were not significantly different comparing both routes. CONCLUSIONS: IV and IO routes produced similar antimicrobial perfusion of the medullary cavity of P1. CLINICAL RELEVANCE: Either IV or IO routes for perfusion are likely to be equally selected when planning RLP with vancomycin. PMID- 16343151 TI - Subtotal ostectomy of dorsal spinous processes performed in nine standing horses. AB - OBJECTIVE: To describe and evaluate subtotal ostectomy of dorsal spinous processes (DSP) performed in standing horses. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Nine adult horses. METHODS: The diseased portions of the DSP were removed with the horses standing and sedated after infiltrating a local anesthetic agent around the affected DSP. Medical records of affected horses were reviewed to determine history, findings of physical and radiographic examination, surgical technique, postoperative complications, and outcome. RESULTS: Five horses had subtotal ostectomy of DSP because of osteomyelitis, and 4 because of impingement and/or fracture of DSP. Affected portions of DSP were removed safely and effectively, with no apparent discomfort to the horses; hemorrhage was minimal. CONCLUSIONS: Subtotal ostectomy of DSP can be performed safely in standing, conscious horses and the risks associated with general anesthesia are eliminated. Hemorrhage is minimal compared to that which occurs when the procedure is performed with horses anesthetized and in lateral recumbency. CLINICAL RELEVANCE: Access and visibility are better when subtotal ostectomy of the DSP is performed with the horse standing rather than anesthetized and in lateral recumbency. Performing subtotal ostectomy of diseased DSP with the horse standing avoids the expense and risks of general anesthesia. PMID- 16343152 TI - Evaluation using hoof wall strain gauges of a therapeutic shoe and a hoof cast with a heel wedge as potential supportive therapy for horses with laminitis. AB - OBJECTIVE: To evaluate using strain gauges, a hoof cast with heel wedge, and a therapeutic shoe with unsupported toe for their effectiveness in redistribution of load from the dorsal hoof wall. STUDY DESIGN: In vitro biomechanical study. SAMPLE POPULATION: Twenty forelimb specimens. METHODS: Rosette strain gauges were placed on the dorsal and lateral hoof wall of 20 normal shaped hooves. Limbs were loaded vertically using a tensile testing machine with a 1 Hz sinusoidally cycling load up to 3000 N during 15 seconds. Mean values of principal strain and direction at 2500 N load were calculated for 3 experimental conditions (unshod, therapeutic shoe with unsupported toe, and hoof cast with heel elevation) and tested by ANOVA (P<.05). RESULTS: Vertical limb loading in an unshod hoof leads to a biaxial compression of the dorsal wall with high longitudinal compression (epsilon2 = -1515 microm/m). Principal strain at the dorsal wall (epsilon2) was decreased by 23% with the therapeutic shoe and by 59% with the hoof cast. On the lateral hoof wall principal strain was unchanged with the shoe, but increased by 34% with the cast. CONCLUSIONS: Strain measurements indicate unloading of the dorsal hoof wall by both methods with the cast being more effective than the shoe. CLINICAL RELEVANCE: The hoof cast with wedge offers substantial unloading of the dorsal wall, but increases load on the quarter. Therefore a hoof cast would likely be most helpful in acute laminitis when palmar structures can still bear load. The therapeutic shoe offers rehabilitation and regrowth of the dorsal wall without increased load on the quarter wall. PMID- 16343153 TI - Indications, surgical technique, and long-term experience with laparoscopic closure of the nephrosplenic space in standing horses. AB - OBJECTIVE: To describe surgical technique and outcome after laparoscopic closure of the nephrosplenic space for treatment of recurrent left dorsal displacement of the left colon (LDDLC) in standing horses. STUDY DESIGN: Retrospective study. ANIMALS: Forty-four horses with LDDLC. METHODS: Laparoscopic portals were located in the left flank region. After administration of etilefrin intravenously to contract the spleen, the dorsal splenic margin was sutured to the nephrosplenic ligament to obliterate the nephrosplenic space. Horses were re-examined (< or = 3 years) for history of recurrence. In addition, medical records of 4,852 horses treated for colic over 16 years were evaluated to establish incidence of LDDLC and recurrence after treatment. RESULTS: Splenic contraction facilitated suture placement and closure of the nephrosplenic space was achieved without complications. LDDLC recurrence did not occur although 5 horses had subsequent episodes of colic; 4 horses had displacement of the ascending colon between the spleen and body wall. Review of medical records revealed an incidence of LDDLC of 6% and recurrence of 21% in a population of horses with signs of colic. CONCLUSION: Laparoscopic closure of the nephrosplenic space, facilitated by etilefrin-induced splenic contraction can be efficiently performed in standing horses. CLINICAL RELEVANCE: Laparoscopic ablation of the nephrosplenic space should be considered in horses that are predisposed to recurrent LDDLC. PMID- 16343154 TI - Laparoscopic closure of the nephrosplenic space for prevention of recurrent nephrosplenic entrapment of the ascending colon. AB - OBJECTIVE: To evaluate outcome after laparoscopic closure of the nephrosplenic space in horses that had previous nephrosplenic entrapment of ascending colon (left dorsal displacement of the left colon; LDDLC). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Horses that had previous LDDLC. METHODS: Medical records of horses that had LDDLC and subsequent laparoscopic nephrosplenic space closure between 2002 and 2004 were retrieved. Follow-up information was obtained by telephone interview of owners. Preoperative versus postoperative comparisons were: incidence of LDDLC, incidence of colic signs, and incidence of ventral celiotomy. Data were analyzed using a chi2-square test with significance set at P<.05. RESULTS: Ten horses met the inclusion criteria. Mean follow-up was 22 months. No horses had recurrence of LDDLC; however, 3 horses had colic signs subsequently that required surgical intervention. Over the follow-up period there was a significant decrease in the total incidence of colic and ventral celiotomy. Technique modifications included use of different cannula site locations, use of polyglyconate suture material for nephrosplenic space closure, and development of a custom cannula. CONCLUSIONS: Laparoscopic nephrosplenic space closure prevented recurrence of LDDLC, and significantly lowered the overall incidence of colic and ventral celiotomy. The custom-designed cannula provided good access to the operative site and no complications were encountered with use of polyglyconate suture material for nephrosplenic space closure. CLINICAL RELEVANCE: Closure of the nephrosplenic space will prevent LDDLC; however, other surgical lesions can occur. PMID- 16343155 TI - Laparoscopic ovariectomy using the endo-GIA stapling device and endo-catch pouches and evaluation of analgesic efficacy of epidural morphine sulfate in 10 mares. AB - OBJECTIVE: To evaluate use of ENDO-GIA staples and ENDO-Catch pouches for ovariectomy in mares and to evaluate the efficacy of epidural morphine analgesia. STUDY DESIGN: Randomized clinical trial. ANIMALS: Mares (n = 10) with normal ovaries. METHODS: An Endo-GIA II stapler (United States Surgical Corp., Norwalk, CT) was used to amputate the ovaries bilaterally, in standing mares, and Endo Catch II pouches (United States Surgical Corp.) were used for retrieval. Epidural morphine was used in 5 mares, and procedure duration, volume of lidocaine required to desensitize the ovarian pedicle, and sedation were compared with horses not administered epidural morphine. RESULTS: Use of the Endo-GIA II stapling device was an easy and an efficient method of ovariectomy, which was facilitated by use of epidural morphine. For the epidural group, the mean (+/-SD) surgical time was 69.2 +/- 10.2 minutes compared with 124.4 +/- 21.2 minutes for mares not administered morphine. Less systemic sedation was required for mares administered epidural morphine. CONCLUSIONS: Minimal ovarian manipulation was required to apply the stapler, hemorrhage was slight, and none of the mares had complications. Epidural morphine provided effective analgesia, improving patient comfort, and reduced the sedation needed to perform ovariectomy. CLINICAL RELEVANCE: Endo-GIA II staples are an efficient method for ovariectomy in normal mares. Epidural morphine should be considered to facilitate ovariectomy because less sedation and local anesthetic administration is required. PMID- 16343156 TI - Laparoscopic ovariectomy in mares using a polyamide tie-rap. AB - OBJECTIVE: To describe use of a polyamide tie-rap to ligate the mesovarium during standing laparoscopic ovariectomy in mares. STUDY DESIGN: Prospective study. ANIMALS: Ten mares. METHODS: Bilateral ovariectomy was performed in 10 mares. Standing laparoscopic ovariectomy was performed using 3 portals in the paralumbar fossa. A commercial polyamide tie-rap was prepared as a loop and marked with 4 colored lines close to the buckle, to enable us to check whether the loop was tightened securely. The ovary was grasped with forceps and after the mesovarium was minimally transected cranially and caudally, the loop of the tie-rap was inserted in the abdomen and placed around the mesovarium. It was firmly tightened, until at least 3 of the 4 marks were visible, then the end of the tie rap was cut. The ovary was transected and removed through an enlarged 3rd portal. The contralateral ovary was removed similarly through the opposite paralumbar fossa. Repeat laparoscopy was performed in 8 mares, 2, 3, 4, and 12 weeks later. RESULTS: None of the mares had postoperative discomfort. On repeat laparoscopy, there was complete encapsulation of the stump and tie-rap after 3-4 weeks. In 2 mares, an adhesion between the left stump and the mesentery of the descending colon was observed. CONCLUSION: Ligation of the mesovarium can be easily and safely performed using a polyamide tie-rap during standing laparoscopic ovariectomy in mares. CLINICAL RELEVANCE: Standing laparoscopic ovariectomy using a polyamide tie-rap is a safe, technically easy and reliable surgical procedure in the mare. PMID- 16343157 TI - Bilateral ureterocystostomy to correct left ureteral atresia and right ureteral ectopia in an 8-month-old standardbred filly. AB - OBJECTIVES: To report the diagnosis and outcome after surgical correction of bilateral distal ureteral anomalies in a Standardbred filly. STUDY DESIGN: Clinical case report. ANIMAL: An 8-month-old, 310 kg Standardbred filly with left ureteral atresia and right ureteral ectopia. METHODS: The filly was admitted for evaluation of incontinence since birth and severe urine scalding of the hindquarters. Diagnosis was made by both direct (cystoscopy and vaginoscopy) and indirect (intravenous pyelography, ultrasonography, and scintigraphy) evaluation of the ureters and bladder. The filly had left ureteral atresia, hydronephrosis, and decreased left-sided renal function and right ureteral ectopia before surgery. Surgical correction was performed on the left by an end-to-side stapled anastomosis technique and on the right by a side-to-side hand-sewn anastomosis technique. RESULTS: Surgical correction was successful. The filly had no postoperative complications and remained continent 18 months after surgery. Left renal function improved. CONCLUSION: Ureteral anomalies can be successfully repaired in larger (>300 kg) foals and some renal function may be restored after surgical correction. CLINICAL RELEVANCE: Scintigraphy should be considered in diagnosis of ureteral anomalies, assessing renal function, and determining prognosis for horses with hydronephrosis caused by ureteral ectopia and atresia. PMID- 16343158 TI - Treatment of a second carpal bone fracture by removal under ultrasonographic guidance in a horse. AB - OBJECTIVE: To report an unusual 2nd carpal bone (C2) slab fracture and its treatment, using an ultrasonographically assisted, minimally invasive arthrotomy technique, for fragment removal. STUDY DESIGN: Case report. ANIMALS: An 11-year old crossbred Warmblood gelding. METHODS: A bone fragment, spanning the height of C2, identified on the palmaromedial aspect of C2 was demarcated intraoperatively using ultrasonographically guided needles and removed by arthrotomy. RESULTS: Fracture fragment removal was achieved using a small, ultrasonographically guided arthrotomy. The horse has become sound after treatment and returned to the previous level of pleasure riding activity. CONCLUSIONS: Perioperative ultrasonography was used to delineate and facilitate removal of an unusual slab fragment of C2, through a minimally sized arthrotomy. Despite the presence of pancarpal osteoarthritis and partial involvement of the medial collateral ligament, the prognosis appeared to be satisfactory for this pleasure riding horse. CLINICAL RELEVANCE: Ultrasonographic guidance may be used to optimize the approach to C2 articular bony fragments that may not be observed using arthroscopy. PMID- 16343159 TI - A dorsal approach for the removal of the nasal septum in the horse. AB - OBJECTIVE: To describe and evaluate a technique involving a dorsal approach for the removal of the nasal septum in adult horses. STUDY DESIGN: Retrospective clinical study. ANIMALS: Four client-owned horses. METHODS: Access for partial nasal septum resection was through a dorsal nasal bone flap. Septum resection was performed because of reduced airflow from septal deviation and/or thickening caused by traumatic insult or neoplasia. Preoperative clinical signs and diagnostic results were recorded. Intra- and postoperative complications were noted and follow-up information was obtained from telephone interviews of owners or by clinical examination. RESULTS: The abnormal segment of the nasal septum was completely removed in all horses. A dorsal nasal bone flap approach provided excellent access to the septum and allowed immediate control of hemorrhage; blood transfusion was not needed. Cosmetic results were considered good to excellent by owners and horses returned to their previous level of work without perceived limitations associated with airway function. CONCLUSIONS: Nasal septum resection by the dorsal approach permits good observation of the abnormal septum, and provides exposure that facilitates removal of nasal masses and reduction of depression fractures with minimal complications. CLINICAL RELEVANCE: A dorsal nasal bone flap approach should be considered for horses that require nasal septum resection because it can be achieved with minimal hemorrhage, good cosmetic results, and return to function. PMID- 16343160 TI - Comparison of left atrial dimensions by transesophageal and transthoracic echocardiography. AB - Transesophageal echocardiography (TEE) is an established cardiovascular diagnostic technique. Left atrial (LA) size, as measured by transthoracic echocardiography (TTE), is associated with cardiovascular disease and is a risk factor for atrial fibrillation, stroke, death, and the success of cardioversion. Assessment of LA size has not been as well validated on TEE as on TTE. We determined LA size measurements in four standard views in 122 patients undergoing TEE and TTE at the same setting. In this study, we found that measurement of LA dimensions by TEE suffers from significant limitations in all views except the basal long-axis view (mid-esophageal level) with transducer plane at 120-150 degrees. This view had the best correlation with transthoracic LA measurements: r = 0.79 for TEE long axis (CI 0.71-0.85), P <.0001. PMID- 16343161 TI - Regional response of myocardial acceleration during isovolumic contraction during dobutamine stress echocardiography: a color tissue Doppler study and comparison with angiocardiographic findings. AB - BACKGROUND: Color tissue Doppler imaging permits noninvasive quantitation of regional wall motion. In experimental studies, a new marker, the slope of the isovolumic contraction wave, isovolumic acceleration (IVA) was more insensitive to ventricular loading conditions than myocardial velocities. This study compared the regional response IVA to dobutamine stress echocardiography to angiographic findings. METHODS: The Myocardial Doppler in Stress Echocardiography (MYDISE) study prospectively recruited 149 consecutive patients with chest pain for dobutamine stress tissue Doppler echocardiography prior to coronary angiography. This color tissue Doppler database was analyzed for IVA in 1,192 basal and mid segments at rest and again at peak stress. Angiographic findings were compared to IVA and peak systolic velocities (PSV) in corresponding cardiac segments. The diagnostic accuracy of IVA to predict coronary artery stenosis was determined using cut-off values for three representative segments and with the MYDISE diagnostic model including eight segments. RESULTS: Regional IVA increased in a dose-dependent manner during dobutamine infusion. The response was blunted in the supply territory of stenosed coronary artery branches. IVA performed slightly better than PSV as single marker for coronary artery stenosis. A diagnostic model incorporating IVA and PSV was 85-95% accurate (area under receiver operating characterstic curves). CONCLUSIONS: Regional changes of isovolumic acceleration during dobutamine stress echocardiography reflect regional wall motion and can be used to predict coronary artery stenosis with similar accuracy as a model based on systolic myocardial velocities. As a single marker, IVA performed better than myocardial velocities. PMID- 16343162 TI - Transesophageal echocardiography using cypress-miniaturized echocardiogram unit: initial clinical experience. AB - Transesophageal echocardiography (TEE) was introduced clinically in the United States in 1987. Recent technologic advances have resulted in the creation of a small portable hand-carried ultrasound (HCU) device that can be easily carried throughout the hospital with greater flexibility for cardiac imaging. These HCU devices have harmonic, color, and spectral Doppler (continuous/pulsed wave). Siemens Medical Solutions USA, Inc. has incorporated a TEE connector, which connects to its Cypress (highly miniaturized echocardiogram unit) and allows the performance of a TEE with this unit, which is mildly heavier than a typical HCU. We describe our initial clinical experience with this unit. The image quality is comparable to routine TEEs, with the advantages of shorter duration, portability, affordable cost, avoiding the use of high-end machine from the echo lab, availability of non-HCU units for other studies, and preventing the need for an echo technician to be involved in the procedure. PMID- 16343163 TI - Characteristics of the interatrial communication in patients undergoing transcatheter device closure of atrial septal defects for cryptogenic stroke. AB - BACKGROUND: Prior studies suggest that patent foramen ovale (PFO) diameter >4 mm is associated with a high probability of cryptogenic ischemic stroke (CIS). METHODS: We evaluated all patients diagnosed with CIS who underwent closure of intra-atrial communication (IAC) using the Amplatzer atrial septal defect (ASD) occluder in our institution between August 1997 and March 2004. For each IAC, echocardiographic diameters and balloon-stretched diameters were recorded. Stretchability index was calculated as the ratio of stretched diameter to unstretched diameter. RESULTS: Fifty-six patients met the inclusion criteria for this study. There was an inverse logarithmic relationship between unstretched IAC diameter and stretchability index. For the 28 smaller defects, the median IAC diameter was 2 mm, and median stretchability index was 5.58 (range 2.6-15). For the 28 larger defects, median diameter was 6 mm, and median stretchability index was 2.38 (range 1.05-5). The difference in stretchability index between the two groups was significant (P < 0.0001). CONCLUSION: Our data bring into question the concept that the diameter of the defect would singularly predict the probability of stroke. PMID- 16343164 TI - Intravenous versus intracoronary myocardial contrast echocardiography for evaluation of no-reflow after primary percutaneous coronary intervention. AB - OBJECTIVE: We sought to compare intravenous myocardial contrast echocardiography (IV-MCE) with intracoronary myocardial contrast echocardiography (IC-MCE) in detecting no-reflow and predicting the short-term outcome of left ventricular function after primary percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI). METHODS: IC-MCE and IV-MCE were performed immediately after PCI (D1) of 28 patients with anterior wall AMI. IV-MCE was repeated at the next day of PCI (D2), and left ventricular systolic function was evaluated at D2 and 30 days later (D30). RESULTS: There was good agreement between IC-MCE and IV-MCE at D1 in determining no-reflow (kappa= 0.78, P < 0.001) as well as between IV-MCE at D1 and D2 (kappa= 0.93, P < 0.001). The patients with no-reflow on IC-MCE (n = 13) and those on IV-MCE at D2 (n = 11) showed no improvement in left ventricular ejection fraction (LVEF) after 1 month (49 +/- 9% to 48 +/- 7%, P = 0.55, and 51 +/- 6% to 49 +/- 7%, P = 0.20). However, the patients with reflow on IC-MCE (n = 15) and those on IV-MCE at D2 (n = 17) demonstrated significant improvement in LVEF (55 +/- 6% to 62 +/- 5%, P < 0.005, and 53 +/- 7% to 60 +/- 8%, P < 0.005). In predicting segmental functional recovery after 1 month, sensitivity and specificity of IC-MCE were 85% and 67%, respectively, and those of IV-MCE at D2 were 95% and 40%, respectively. CONCLUSION: IV-MCE at D2 might be substituted for IC-MCE performed immediately after PCI for the evaluation of no-reflow and prediction of left ventricular systolic function after 1 month in patients with anterior wall AMI. PMID- 16343165 TI - Effect of cardiac resynchronization therapy on longitudinal and circumferential left ventricular mechanics by velocity vector imaging: description and initial clinical application of a novel method using high-frame rate B-mode echocardiographic images. AB - Cardiac resynchronization therapy (CRT) has emerged as an important method to treat patient with symptomatic heart failure with evidence of intraventricular dyssynchrony. Tissue Doppler imaging by echocardiography has been shown to be an excellent tool for the assessment of mechanical left ventricular dyssynchrony and the selection of patients for CRT. However, there are some patients who do not show symptomatic improvement following CRT. One possible explanation for this is the need to optimize not only longitudinal synchrony, but also improve the circumferential and radial dynamics of the left ventricle. Doppler imaging does not allow reliable assessment of the latter because of the angle-dependency of the technique. Velocity Vector Imaging (VVI) is a newer technique which is angle independent and thus provides an avenue to evaluate short-axis mechanics of the left ventricle. We describe a case in which VVI was used to assess the left ventricular dynamics in a patient with heart failure who did not respond to CRT. PMID- 16343166 TI - Contrast-enhanced echocardiography for the diagnosis of apical hypertrophic cardiomyopathy. AB - The echocardiographic diagnosis of apical hypertrophic cardiomyopathy can be difficult in patients with poor acoustic windows. However, contrast-enhanced echocardiography can provide better images in these patients and lead to the correct diagnosis. We present a patient with apical hypertrophic cardiomyopathy who was diagnosed using contrast-enhanced echocardiography. The use of contrast enhanced echocardiography for the diagnosis of apical hypertrophic cardiomyopathy in patients with poor acoustic windows is discussed and the experience in the literature reviewed. PMID- 16343167 TI - Rapid diagnosis and management of intraoperative myocardial infarction during valvular surgery: using intraoperative transesophageal echocardiography followed by emergency coronary artery bypass grafting without coronary angiography. AB - A 68-year-old man was admitted to undergo elective mitral valve surgery. Although the preoperative coronary angiography was normal, the patient suffered a myocardial infarction that resulted in untreatable collapsed hemodynamics. After inferring the responsible occluded coronary artery from the segmental wall motion abnormality detected in intraoperative transesophageal echocardiography, together with the anatomy found in preoperative coronary angiography, we performed an emergency coronary artery bypass graft surgery without a new angiography. This procedure resulted in survival of a potentially life-threatening situation. In selected cases, this therapeutic strategy may lead to reduction of mortality as a result of the intraoperative myocardial infarction. PMID- 16343168 TI - Cor triatriatum with single atrium presenting in adulthood. AB - We present the case of an asymptomatic 21-year-old woman referred because of an abnormal routine electrocardiogram. Transthoracic and transesophageal echocardiography revealed a complete absence of the atrial septum, a common atrium septated posteriorly from the pulmonary venous chamber, a partial atrioventricular canal, a cleft mitral valve and a persistent left superior vena cava draining into an enlarged coronary sinus. These findings were confirmed during surgical correction. Our patient presented with an unusual and fascinating combination of congenital malformations that remained well tolerated and undiscovered into adulthood. PMID- 16343169 TI - Rupture of a right sinus of valsalva aneurysm into the right ventricle during vaginal delivery: a case report. AB - A case is reported of a right sinus of Valsalva aneurysm rupture into the right ventricle during vaginal delivery in a 34-year-old healthy woman in her third pregnancy. Pregnancy was carried to term and a healthy baby was delivered vaginally. On day 7 following vaginal delivery she was admitted to hospital for dyspnea and cough, with clinical signs of severe heart failure. The diagnosis of the right sinus of Valsalva aneurysm rupture into the right ventricle was established by transthoracic and transesophageal echocardiography. Clinical recognition and early echocardiographic diagnosis followed by immediate surgical repair proved lifesaving in our patient. PMID- 16343170 TI - Live/real time three-dimensional transthoracic echocardiographic assessment of Ebstein's anomaly. AB - We describe our experience in using live/real time three-dimensional transthoracic echocardiography (3D TTE) in the assessment of five adult patients with Ebstein's anomaly. The technique was found useful in assessing the distribution and extent of tethering of each of the three leaflets of the tricuspid valve (TV) to the underlying right ventricular walls and the ventricular septum. The characteristic bubble-like appearance resulting from bulging of the non-tethered areas of the TV leaflets was also well visualized in three dimensions and their size measured. Thus, an estimate of the nontethered or free segments of all three leaflets of the TV could be obtained using this technique. This has important implications when considering these patients for surgical repair of the TV. Visualization of all three leaflets of the TV and their extent of tethering by 3D TTE also made it easier to identify the boundaries of the functioning right ventricular chamber potentially providing a more reliable assessment of its volumes and ejection fraction. Cropping of color Doppler 3D TTE data sets provided en face viewing of the TV regurgitation vena contracta permitting accurate assessment of its shape and size. This has the potential to provide a more accurate quantitative estimation of TV regurgitation severity as compared to two-dimensional color Doppler. In conclusion, live/real time 3D TTE appears useful in supplementing two-dimensional echocardiography in more comprehensively assessing the morphologic features of Ebstein's anomaly. PMID- 16343171 TI - Asymptomatic left ventricular myxoma diagnosed incidentally by transthoracic echocardiography. PMID- 16343172 TI - Membranous septal aneurysm causing severe tricuspid regurgitation and ring-like appearance. PMID- 16343173 TI - Aortic root involvement in homozygous familial hypercholesterolemia- transesophageal echocardiographic appearances of supravalvular aortic stenosis. PMID- 16343174 TI - Echocardiographic findings in HIV Disease and AIDS. PMID- 16343175 TI - Radical hysterectomy for stage IIB cervical cancer: a review. AB - Patients with stage IIB cervical cancer in some countries in Europe and Asia especially in Japan are usually treated with radical hysterectomy and pelvic lymphadenectomy. Extrauterine diseases, ie, nodal metastases, parametrial invasion, and intraperitoneal spread, can be readily identified. We present the literature review of radical hysterectomy in stage IIB cervical cancer by searching data since 1980 from Medline, and we found that the parametrial involvement of patients in this stage was only 21-55%, the incidence of pelvic node metastases was about 35-45%, and 5-year survival rate was between 55% and 77%. Lymph node metastases and the number of positive nodes were significant prognostic factors of patients in this stage. PMID- 16343176 TI - Gemcitabine in epithelial ovarian cancer treatment: current role and future perspectives. AB - Newer agents and combinations are needed in order to improve current results in ovarian cancer treatment. Gemcitabine is a novel agent that has shown consistent activity as a single agent in the treatment of platinum-resistant ovarian cancer and a favorable toxicity profile. Because of its clinical and preclinical synergism with platinum analogs, gemcitabine has been combined with carboplatin as a convincing approach in the treatment of platinum-sensitive recurrent ovarian cancer patients. Further combination of gemcitabine and other agents, including paclitaxel, is also feasible and has been actively studied in order to establish the role of gemcitabine in the management of treated and untreated ovarian cancer patients. PMID- 16343177 TI - Prognostic factors in ovarian carcinoma stage III patients. Can biomarkers improve the prediction of short- and long-term survivors? AB - The aim of the study was to determine if biomarker expression could help discriminate between short-term and long-term survivors in women with advanced ovarian cancer. Fifty-one patients with stage III ovarian cancer were selected for the study, which included 28 short-term survivors (death from ovarian cancer within 18 months) and 23 long-term survivors (alive for more than 5 years). There was no difference between the two groups with respect to FIGO substage, age, World Health Organization score, and first-line platinum therapy. Classic clinical pathologic parameters were examined together with p53, Bcl-2, Ki-67, PDGFRalpha, P-glycoprotein, BRCA1, and DNA ploidy. Immunohistochemistry was used for scoring biomarker expression and image cytometry for DNA ploidy. All patients had primary debulking surgery followed by first-line platinum therapy. On multivariate analysis, the presence of ascites, debulking surgery and repeat laparotomy, clear-cell histology, elevated CA125, and high Ki-67 score were all found to be of prognostic importance. The long-term survivors were characterized by primary optimal cytoreduction surgery (<1 cm residual disease), attempt at maximal tumor debulking by experienced gynecological oncologic surgeons, and the absence of ascites. Normal CA125 level before platinum therapy and negative Ki-67 expression also predicted a more favorable prognosis. PMID- 16343178 TI - A pilot phase 2 study of oregovomab murine monoclonal antibody to CA125 as an immunotherapeutic agent for recurrent ovarian cancer. AB - This prospective, open-label, pilot phase 2 study examined the clinical and immunologic effects of oregovomab (OvaRex) in heavily pretreated patients with recurrent ovarian cancer (OC). Thirteen women were administered intravenous oregovomab (2 mg) at weeks 0, 2, 4, 8, and 12, followed by quarterly doses for up to 2 years or disease progression. Concomitant chemotherapy was not permitted. Eligibility criteria included recurrence after one or more platinum-based chemotherapy regimens, CA125 >35 U/mL, evaluable or measurable disease. Tumor burden was evaluated by physical or radiologic methods pretreatment, weeks 12, 24, and every 24 weeks thereafter. Immune responses, including antibodies and T cells to oregovomab and CA125, were demonstrated in over half the patients. Stabilization of disease and survival >2 years was observed in 3 of 13 patients and coincided with robust immune responses. Shrinkage of marker lesions was not observed; however, four patients showed decreases in CA125 levels. Treatment was well tolerated without serious adverse events or discontinuations due to therapy. This pilot study supports immunologic activity and safety of oregovomab in recurrent OC. Further study of this agent in the consolidation and adjuvant setting is ongoing to establish its clinical utility. PMID- 16343179 TI - A phase II study of fixed dose rate gemcitabine in patients with relapsed mullerian tumors. AB - Gemcitabine (2',2'-difluorodeoxycytidine) is a novel purine analog with clinical activity against ovarian cancer. Accumulation of gemcitabine triphosphate (dFdCTP) increases in a linear fashion with prolonged infusions of gemcitabine, and there is a strong relationship between intracellular accumulation of dFdCTP and DNA damage. Women with ovarian, fallopian tube, or primary peritoneal carcinoma and documented recurrent disease were eligible for the study. Patients could not have received more than four prior lines of chemotherapy and had to have measurable or evaluable disease. Gemcitabine 800 mg/m2 administered by intravenous infusion at 10 mg/m2/min (fixed dose rate [FDR]) on days 1 and 8 of a 21-day schedule. Twenty-eight patients with a median age 60 (range, 40-77) years were treated. Although 43% were Eastern Cooperative Oncology Group 0, 50% had liver metastases. Eighty-eight cycles of therapy were delivered (median 2 [range, 1-6]). Five of the first ten patients treated at 800 mg/m2 could not receive day 8 FDR-gemcitabine because of neutropenia, and the starting dose was reduced to 700 mg/m2. Even at this dose there was cumulative hematologic toxicity resulting in dose reductions. Vomiting, mucositis, diarrhea, allergy, rash, fever, and alopecia were mild. In 28 patients, there was only one partial response (4%, 95% CI 0-18%) and median time to progression was 1.7 (interquartile range, 1.2-3.9) months. FDR-gemcitabine 700 mg/m2 administered by intravenous infusion at an FDR of 10 mg/m2/min had minimal activity against heavily pretreated recurrent tumors of mullerian origin. The optimal dose and schedule of gemcitabine is yet to be defined in this population. PMID- 16343180 TI - Breast cancer resistance protein-mediated topotecan resistance in ovarian cancer cells. AB - Overexpression of breast cancer resistance protein (BCRP) and mitoxantrone (MX) resistance protein can confer resistance to a variety of cytostatic drugs, such as MX, topotecan (TPT), doxorubicin, and daunorubicin. This study investigates the role of BCRP in resistance of ovarian cancer to TPT treatment. We have developed TPT-resistant human ovarian cancer cell line. Intracellular concentration of fluorescent dye rhodamine 123 (Rh123) was measured by flow cytometry. The expression of several membrane transporter proteins including P glycoprotein (P-gp), multidrug resistance protein 1 (MRP1), and BCRP were determined by reverse transcription-polymerase chain reaction and Western blotting. The Rh123 concentration in parental cells was approximately three times of those in TPT-resistant cells. In contrast to undetectable level of P-gp messenger RNA (mRNA) and minimal level of MRP1 expression in TPT-resistant cells, overexpression of both the BCRP mRNA and the protein was detected in these cells. Introduction of antisense-phosphorothioate oligonucleotide derived from BCRP mRNA into TPT-resistant cells resulted in a significant increase in the concentration of intracellular Rh123. These results suggested a novel mechanism in which a reduced intracellular drug concentration may be mediated by BCRP gene products in human ovarian cancer cells. PMID- 16343181 TI - An ethnographic study of cervical cancer among women in rural Kenya: is there a folk causal model? AB - This article assesses knowledge, attitudes, and practices regarding cervical cancer among rural women of Kenya. One hundred and sixty women (mean age 37.9 years) who sought various health care services at Tigoni subdistrict hospital, Limuru, Kenya, were interviewed using a semistructured questionnaire. In addition, three focus group discussions (25 participants) were held, five case narratives recorded, and a free list of cervical cancer risk factors obtained from a group of 41 women respondents. All women were aged between 20 and 50 years. About 40% knew cervical cancer, although many still lack factual information. A history of sexually transmitted diseases (61.5%), multiple sexual partners (51.2%), and contraceptive use (33%) were identified as risk factors. Other factors mentioned include smoking, abortion, and poor hygiene standards. High parity, early sexual debut, and pregnancy were not readily mentioned as risk factors. We propose a folk causal model to explain the link between these factors and cervical cancer. Lack of knowledge constrains utilization of screening services offered at the clinics. Consequently, respondents support educating women as a way to tackling issues on cervical cancer. It is recommended that an integrated reproductive health program that addresses comprehensively women's health concerns be put in place. PMID- 16343182 TI - Whole-body [18F]fluoro-2-deoxyglucose positron emission tomography scan staging prior to planned radical hysterectomy and pelvic lymphadenectomy. AB - The aim of the study was to determine the effectiveness of whole-body [18F]fluoro 2-deoxyglucose positron emission tomography (FDG PET) imaging in properly selecting candidates for radical hysterectomy who are at low risk for subsequent chemoradiation. Retrospective study of 14 women undergoing planned radical hysterectomy and pelvic lymphadenectomy with clinically localized cervical cancer and either negative or inconclusive metastatic nodal disease by PET scan was performed. Pelvic lymph nodes were clearly negative by FDG PET scan in 12 of the 14 women. Two women had focal FDG uptake suspicious, although not definitive, for nodal metastasis, and pelvic nodes were positive at surgery in both. Computed tomography (CT) scan failed to detect nodal disease in either woman. Neither PET nor CT was effective at detecting parametrial disease, and both also failed to detect the primary tumor in some cases. Women with FDG PET scans that are clearly negative for nodal disease are good candidates for radical hysterectomy and are at low risk for subsequent chemoradiation. PMID- 16343183 TI - The clinicopathologic significance of laminin-5 gamma2 chain expression in cervical squamous carcinoma and adenocarcinoma. AB - Carcinoma of the uterine cervix is one of the most prevalent malignancies among women in developing countries and the third most common type worldwide. Squamous cell carcinoma predominates in the cervix uteri, while adenocarcinoma and adenosquamous carcinomas represent about 10-15% of all cervical cancers. Many studies have confirmed that the human papillomavirus (HPV) is the most important etiologic factor in the development of cervical cancer. The aim of our study was to investigate the expression of the laminin-5 gamma2 chain in primary malignancies of the cervix uteri and to focus on the clinicopathologic significance of the expression of the laminin-5 gamma2 chain in cervical squamous carcinoma and adenocarcinoma with respect to age and survival of the patients. The study consisted of a total of 89 cases of invasive cervical cancer (54 squamous carcinomas and 35 adenocarcinomas). The laminin-5 gamma2 chain was found in 80% of all the squamous carcinoma and in 66% of cervical adenocarcinoma. There was no correlation of the high expression of laminin-5 with survival. The univariate analysis in squamous cell carcinoma showed that factors such as the stage of the disease and positive lymph nodes had an impact on the survival of the patients, whereas in the multivariate analysis, only age at diagnosis was an independent prognostic factor. However, in cases with cervical adenocarcinoma, only the stage of the disease was an independent prognostic factor. There was no difference between HPV-positive and HPV-negative tumors concerning the high expression of laminin-5 gamma2 chain. Our results indicate that the majority of the primary cervical tumors, especially squamous cell carcinoma, showed expression of laminin-5 gamma2 chain immunoreactivity. Independent prognostic values for the survival of the patients were age and stage of the disease. PMID- 16343184 TI - Expression of inhibitory natural killer receptors on tumor-infiltrating CD8+ T lymphocyte lineage in human endometrial carcinoma. AB - To investigate the expression of natural killer receptors (NKRs) within the human tumor milieu, we directly examined the in vivo expressions of various NKRs on tumor-infiltrating lymphocytes (TILs) derived from human endometrial carcinoma (EC). In total, 22 patients with stage IA-IIIA EC were enrolled. TILs were isolated from tissue specimens by means of a mechanical dispersal technique. The subpopulations of immunocytes were quantified, and expressions of NKRs on CD8+ T cells were analyzed by triple-color flow cytometry. CD8+ T cells express higher ratios of CD94 and NKG2A in TILs than in peripheral blood mononuclear cells (PBMCs) in human EC. Flow cytometry reveals that 15.90% of CD3+CD8+ TILs compared with 2.10% of CD3+CD8+ PBMCs express the NKG2A molecules (P < 0.001). The percentage expressions of CD94 are 8.40% in CD3+CD8+ TILs and 3.80% in CD3+CD8+ PBMCs (P= 0.013). The numbers of CD8+ T cells expressing CD158b and NKB1 are higher in CD3+CD8+ PBMCs in EC than in normal (CD158b: 10.70% vs 2.60%, P < 0.001; NKB1: 2.20% vs 0.40%, P= 0.018, respectively). Increased expression of CD94/NKG2A restricted to tumor-infiltrating CD8+ T cell subsets may shape the cytotoxic responses, which indicate a possible role of tumor escape from host immunity in human EC. PMID- 16343185 TI - FHIT expression in neoplastic, hyperplastic, and normal endometrium. AB - Fragile histidine triad (FHIT), a candidate of tumor suppressor protein, expression was examined on paraffin-embedded specimens in proliferative, secretory, hyperplastic, and neoplastic human endometrium by immunohistochemistry. The results of FHIT immunoreactivity in endometrial carcinomas were compared with prognostic indicators as well as with p53 overexpression. Forty-four cases of endometrial carcinoma, 30 normal functional (15 proliferative, 15 secretory), and 24 hyperplastic endometrium (12 without atypia, 12 with atypia) specimens were studied using polyclonal FHIT antibody. The streptavidin-biotin-peroxidase detection system was used, and the intensity and the distribution of immunoreactivity were evaluated semiquantitatively. There were no significant differences in FHIT expression in the proliferative, secretory, hyperplastic, either with or without atypia, or carcinomatous endometria. No significant difference in FHIT expression of endometrial carcinomas was detected when prognostic parameters or p53 overexpression were considered. Loss or reduced FHIT expression was not found to predict disease-free or cumulative survivals. This study showed that loss or reduction in FHIT protein expression is present in normal functional and hyperplastic endometria as well as in neoplastic endometrium. FHIT protein seems not to be involved directly in endometrial carcinogenesis, but rather, it regulates cell proliferation both in physiologic and in pathologic conditions of endometrium. PMID- 16343186 TI - Variable MLH1 promoter methylation patterns in endometrial carcinomas of endometrioid subtype lacking DNA mismatch repair. AB - A lack of DNA mismatch repair (MMR) is observed in approximately 20% of sporadic endometrial tumors, but few of these tumors have mutations in MSH2 or MLH1, the two major MMR genes. Promoter methylation is an important means of silencing transcription, and methylation of the MLH1 promoter has been reported as an important cause of MLH1 inactivation in endometrial cancers. Studies have shown that specific CpG sites within the MLH1 gene promoter are critical for gene expression, but other studies have shown that methylation of both more proximal and more distal sequences are important for MLH1 gene regulation. Here, we used a microsatellite instability assay and MLH1 immunohistochemistry to identify a subset of endometrial carcinomas of the endometrioid subtype lacking MMR. Sequencing of bisulphite-treated DNA from these tumors determined the methylation status of 42 CpG sites across the MLH1 promoter (spanning -204 to -702 bp upstream of the transcriptional start). Unlike the 4 normal endometrial samples that were unmethylated, 17 of 21 MMR-deficient samples showed complete or near complete methylation and the remaining 4 MMR-deficient samples had a considerable degree of methylation (approximately 50% or greater). Five tumors demonstrated isolated unmethylated CpG sites, despite methylation throughout the rest of the promoter. This underscores the importance of examining the methylation status of at least several CpG sites within the promoter as methylation is not always consistent across DNA. Overall, our findings support the model that density of methylation of CpG sites across the MLH1 promoter is important in determining gene expression. PMID- 16343187 TI - Aggressive angiomyxoma of the perineum. AB - We report the case of perineal aggressive angiomyxoma in a 46-year-old woman presenting as left perineal swelling associated with superficial dyspareunia. Initial clinical examination revealed a 4- to 5-cm cystic mass in the posterior aspect of the left labia majora, not thought to be typical of a Bartholin cyst. A magnetic resonance imaging (MRI) scan revealed a well-defined 2- x 1.5- x 2-cm area posterolateral to the lower vagina on the left but anterolateral to the anal canal extending into the left ischiorectal fossa, with no obvious involvement of the anal sphincter complex. Excision biopsy was performed via an incision in the left labia majus under general anesthesia. Histologic findings were consistent with aggressive angiomyxoma of the vulva. This was confirmed by immunohistochemistry showing spindle cells positive for vimentin with strong nuclear staining for estrogen and progestogen receptors. Postoperative management following discussion at the multidisciplinary gynecological oncology meeting was to perform a repeat MRI scan 6 weeks postoperatively, and treatment with raloxifene was commenced for its antiestrogenic property. PMID- 16343188 TI - Benign multicystic peritoneal mesothelioma: cases reports in the family with diverticulosis and literature review. AB - We report on benign multicystic peritoneal mesothelioma in two siblings whose family had a history of multiple familial diseases including diverticulosis. After a genetic evaluation and a chromosomal analysis, we were not able to identify a specific genetic cause of the family's pattern of disease. We assumed that previous surgical procedures and the chronic inflammatory process from diverticulitis were the underlying etiology. Both patients had multiple recurrences with indolent courses similar to those reported in other cases. After the recurrences, one patient was treated with cystic aspiration and the other with hormones. The cysts in both cases regressed partially but the patients were relieved of their clinical symptoms, for 2 years after cystic drainage in one case and for 5 years after hormonal treatment in the other. PMID- 16343189 TI - Primary undifferentiated carcinoma arising from the retroperitoneum: a case report. AB - A 35-year-old woman, gravida 1, para 1, underwent cesarean section in her 39th week of pregnancy. At the time of operation, multiple retroperitoneal tumors were found. Postoperative computed tomography and magnetic resonance imaging showed multiple solid tumors, each approximately 3-5 cm, in the right pelvic retroperitoneal space. Total resection of the tumors was performed without any macroscopic residual. A systematic workup for the primary tumor from which the retroperitoneal tumors may have metastasized failed to demonstrate any responsible tumor. We therefore assumed it to be a primary retroperitoneal tumor. The histopathologic features of the tumors were consistent with small-cell carcinoma. Two months postoperatively, recurrent tumors in the right inguinal and common iliac regions were detected. Since chemotherapy with irinotecan hydrochloride or paclitaxel did not produce any beneficial effect, a second tumor reduction surgery was performed 8 months after the initial operation. Four months after the second operation, a third operation including total hysterectomy, bilateral salpingo-oophorectomy, and tumor resection in the contralateral iliac region were done. Afterward, a new recurrent tumor appeared along the aorta up to the left supraclavicular node. The patient died 19 months after the first operation. PMID- 16343190 TI - Long-term survival following aggressive surgery and radiotherapy for pelvic fibromatosis. AB - Desmoid tumors of the female pelvis are rare. The efficacy of the available treatment modalities in improving survival and decreasing recurrence remains controversial. A 32-year-old woman presented with an asymptomatic large ischeorectal mass. Computed tomography scan revealed a large tumor adherent to the pubic bone and impinging on the bladder neck and the rectum. Aggressive surgical removal of the mass including partial osteotomy of the pubic bone was followed by radiotherapy. The patient is still alive 6 years later with no evidence of disease. Aggressive surgical management followed by radiotherapy is an acceptable means of treatment of locally invasive desmoid tumor of the female pelvis. PMID- 16343191 TI - Major hepatectomy and right hemicolectomy at the time of primary cytoreductive surgery for advanced ovarian cancer: report of a case. AB - Major liver involvement at the time of diagnosis is a rare event in patients with ovarian cancer, and the issue of major hepatectomy at the time of primary cytoreductive surgery is controversial. A 61-year-old woman was admitted to our hospital with nonspecific abdominal pain of 2-month duration and weight loss of 5 kg during the last semester. A computed tomography scan demonstrated bilateral ovarian masses, extending to the right iliac fossa, pressing the cecum-ascending colon. In the liver parenchyma, three cystic lesions were found of about 6-cm maximum diameter each, along with pelvic lymphadenopathy. There was no ascites. The diagnosis of advanced ovarian cancer was clinically suspected; the patient underwent a total abdominal hysterectomy with bilateral salpingo-oophorectomy, right hemicolectomy, omentectomy, left lobectomy, deroofing, and draining of the cystic formation of the right liver lobe along with systematic pelvic and para aortic lymphadenectomy. Systemic chemotherapy (six cycles of paclitaxel/carboplatin) was subsequently administered, and after 15 months of follow-up period, the patient is still in first remission and alive. Ovarian cancer with concomitant extensive right colon infiltration and hematogenous liver metastases can be successfully managed with aggressive surgical resection and postoperative chemotherapy in carefully selected patients. PMID- 16343192 TI - Ovarian neoplasms and pregnancy. AB - The aim of our study was to investigate the incidence of ovarian malignant neoplasms in pregnancy. In the prospective study, during a 2-year period of time, we had the ethical dilemma concerning the conciliar treatment vs the patient's wish to deliver and save the reproductive capabilities. We examined 37 pregnant women; the incidence of ovarian cancer was 13.5% vs 6.5% in other works. Diagnoses were made by ultrasound criteria and physical examinations. Statistically significant results were obtained by Student t test. Mean gestational age was 20.1 weeks and mean age 31.1 years. Treatment depends on the neoplasms type, grade, and presence of the metastatic pathways. In benign neoplasms, we used laparoscopic treatment with minor invasion. Distribution of benign neoplasms was in the same range as that in other works. PMID- 16343193 TI - Dermatomyositis following the diagnosis of ovarian cancer. AB - We present a case history of a woman who developed dermatomyositis following the diagnosis of stage IV ovarian cancer. Dermatomyositis is a rare paraneoplastic syndrome that usually precedes the diagnosis of ovarian cancer by several months or years. Ours is the fifth reported case of dermatomyositis after an established diagnosis of ovarian cancer in the literature. PMID- 16343194 TI - Positron emission tomography in a premenopausal asymptomatic woman: a case report of increased ovarian uptake in a benign condition. AB - Positron emission tomography (PET) use is increasing; however, optimal utilization in gynecologic oncology remains unclear. PET is expensive, has limited anatomic detail, and it may be difficult to differentiate benign ovarian lesions from malignant lesions when PET is used. A 43-year-old female volunteer's PET scan revealed increased uptake in the left ovary. A subsequent extensive evaluation was entirely normal; however, the patient pursued excision with only a corpus luteum on final pathology. There is a dearth of information regarding PET scan ovarian abnormalities in asymptomatic premenopausal patients, as much of the literature focuses on women with a known ovarian lesion or cancer. Our case represents an increasingly common situation: evaluation and management of an asymptomatic woman with a positive PET scan. As more clinicians encounter PET scan abnormalities, it is imperative that the medical literature documents the limitations of this technology, especially in premenopausal women. PMID- 16343195 TI - Fallopian tube carcinoma presenting as tubo-ovarian abscess: a report of two cases with literature review. AB - Preoperative diagnosis of fallopian tube carcinoma is difficult, with fewer than 5% being diagnosed preoperatively. We describe tubal carcinoma, presenting as a tubo-ovarian abscess in two 47-year-old women. Both patients presented with abdominal pain, pelvic mass, and fever. Both patients were treated as having a tubo-ovarian abscess but failed to respond to therapy. During surgery a metastatic right tubal carcinoma was found. A definite operation was performed in both patients. Three additional cases of fallopian tube carcinoma, presenting as acute pelvic inflammatory disease, were found while reviewing the English literature. Actually all these three cases presented as tubo-ovarian abscess because of the existence of tender pelvic mass. Carcinoma of the fallopian tube should be considered in the differential diagnosis of tubo-ovarian abscess in those who failed to respond to a previously unreported clinical presentation. PMID- 16343196 TI - Giant malignant ovarian tumor: a case report. AB - Massive ovarian tumors are rare and occur less frequently today. The management of a patient with a large ovarian tumor weighing 74 kg is described. Preoperative paracentesis was resorted to. A multidisciplinary team approach helped combat the various associated complications of supine hypotension, technically difficult surgery, massive blood and fluid replacement, postoperative ventilatory failure, and wound dehiscence. PMID- 16343197 TI - Primary ependymoma of the ovary: a case report and literature review. AB - Ependymoma is a glioma with differentiation toward ependymal cells that usually arises in the central nervous system. Ovarian ependymoma is extremely rare, and the treatment strategies for this disease have not been established. This is the first report of a patient with advanced ovarian ependymoma who received fertility sparing surgery and is now alive without disease. A 23-year-old Japanese woman, gravida 0, presented with a chief complaint of lower abdominal distension and weight loss. She underwent left salpingo-oophorectomy for suspected ovarian cancer. Microscopic examination revealed a highly cellular tumor composed of small cells with hyperchromatic, round-to-oval nuclei and scanty cytoplasm. Perivascular pseudorosettes, ependymal rosettes, and extensive necrosis were observed. After thorough pathologic examination, she was diagnosed as having stage IIIC ovarian ependymoma. Postoperatively, she received adjuvant chemotherapy and underwent secondary cytoreductive surgery that preserved the uterus and right ovary. Her menstrual cycle has resumed, and she is alive without evidence of disease 16 months after the start of treatment. Although rare, primary ovarian ependymoma must be kept in mind in the differential diagnosis of ovarian tumors, especially in young women. Administration of etoposide-based chemotherapy along with cytoreductive surgery is a potential standard treatment for advanced ovarian ependymoma. PMID- 16343198 TI - Primary ovarian fibrosarcoma: a case report and review of the literature. AB - Primary ovarian fibrosarcomas are very rare tumors with great heterogeneity among reported cases. There are only a few reports of the subject in the literature, and along with the nature of the disease, diagnosis and treatment still remain to be established. In this report, we reviewed the cases published in the English literature within the past 30 years and tried to highlight certain aspects of the disease. A 52-year-old parous woman was admitted to our hospital with the complaints of abdominopelvic pain. Initial diagnostic work-up revealed a solid mass on the right ovary. After explorative laparotomy, the pathologic examination reported a primary ovarian fibrosarcoma. The patient has been free of disease for 1 year without any adjuvant therapy. Although ovarian fibrosarcomas are unusual causes of solid masses in postmenopausal women, it should be kept in mind when evaluating adnexal masses in this age group. PMID- 16343199 TI - Metastatic malignant melanoma arising in a mature ovarian cystic teratoma: a case report and literature review. AB - Primary malignant melanoma arising inform the ovary is rare, with only 30 cases described in the literature to date. The case reported here occurred in a 19-year old woman and was rapidly progressive, resulting in death only 37 days following initial presentation. Management of this case is discussed in the context of the other reported cases. Surgery, ranging from an ovarian cystectomy to a radical debulking procedure, has been the main treatment with adjuvant chemotherapy utilized in only 4 of the previous cases. Key problems in management relate to the ability to make the diagnosis at the time of surgery and the overall poor response rates of melanoma to adjuvant chemotherapy. PMID- 16343200 TI - Ovarian sex cord-stromal tumor, unclassified: case report of endocrinologic findings and review of the literature. AB - Endocrinologic findings and a successful pregnancy in a 39-year-old woman with significant androgen excess due to a biologically active, virilizing ovarian sex cord-stromal tumor, unclassified, are described. She had secondary amenorrhea, infertility, and hirsutism. Her basal plasma testosterone level was high at 187 ng/mL (normal range: 10-85 ng/mL). Tumor resection resulted in resumption of ovulation, pregnancy, and delivery. PMID- 16343201 TI - Struma ovarii showing clinical characteristics of ovarian malignancy. AB - Struma ovarii is a rare form of ovarian neoplasm, composed entirely or predominantly of thyroid tissue and generally a benign germ cell tumor of the ovary. Ascites have been reported in approximately 15-20% of all cases. However, the combination of struma ovarii and elevated CA125 has rarely been reported. We experienced a rare case of a postmenopausal woman with benign struma ovarii associated with massive ascites, a complex pelvic mass. There was marked elevation of her CA125 level. The clinical impression was ovarian malignancy. Surgical excision of the ovarian mass induced immediate resolution of the ascites and a normalization of the serum CA125 level. No recurrence of the ascites or of the tumor has been observed during the 10-month follow-up. Struma ovarii can mimic ovarian malignancy clinically, particularly if complex and associated with ascites and an elevated CA125 level. PMID- 16343202 TI - Complete remission of an endometrial carcinoma with bilateral multiple pulmonary and extrapelvic metastases treated by surgery and chemotherapy consisting of paclitaxel and carboplatin. AB - Successful management of a patient with endometrioid type, grade 2 endometrial carcinoma with bilateral multiple pulmonary and extrapelvic abdominal metastases has been reported. A 61-year-old woman with the preoperative diagnosis of stage IVB endometrial carcinoma underwent surgery followed by six cycles of chemotherapy consisting of paclitaxel (175 mg/m2) and carboplatin (area under curve 5). After the sixth course, there were no abnormal findings on chest and abdominal computed tomography. She has no evidence of disease recurrence 24 months after the induction of chemotherapy. Tumor markers are within normal limits. Endometrial carcinoma with pulmonary metastases, especially those with bilateral multiple pulmonary metastases associated with additional extrapulmonary spread can be successfully treated by extensive surgery followed by chemotherapy consisting of paclitaxel and carboplatin. PMID- 16343203 TI - A case of postirradiation vaginal angiosarcoma treated with recombinant interleukin-2 therapy. AB - Angiosarcoma of the vagina is an extremely rare neoplasm and is characterized by frequent recurrence and early metastatic spread. Although previous reports emphasized the poor prognosis of this disease, effective treatment strategies have not been adequately stated. We report a case of angiosarcoma of the vagina, in which the diagnosis was made 9 years after intrapelvic irradiation, and recombinant interleukin-2 (rIL-2) therapy could be effective to suppress the development of distant metastasis. We recommend rIL-2 therapy in combination with irradiation as a palliative therapeutic option for vaginal angiosarcoma when the tumor is inoperable or the patient refuses to undergo surgery. Although vaginal angiosarcoma is an extremely rare condition, its possibility should be borne in mind when finding a vaginal mass in a previously irradiated patient. PMID- 16343204 TI - A case of alveolar soft part sarcoma with vaginal metastasis: successful control of vaginal bleeding with external beam irradiation. AB - Alveolar soft part sarcoma (ASPS) of the vagina is an exceptionally rare neoplasm. Furthermore, vaginal metastasis of ASPS has not been reported. A 28 year-old woman with a history of a right thigh mass diagnosed as ASPS excised 8 years ago presented to the emergency room with massive vaginal bleeding and anemia. Biopsy of a vaginal mass revealed that the tumor was a vaginal metastasis of ASPS. For control of intractable bleeding and preventing further transfusions, palliative radiation therapy was planned. She received a total of 39 Gy (daily 3 Gy, using 15-MV photons), and after 6-Gy irradiation, there was no more vaginal bleeding and no more transfusion needed. This is the first case of vaginal metastasis of ASPS reported in the literature that was manifested by intractable vaginal bleeding, which was controlled successfully with radiation therapy. PMID- 16343205 TI - Malignant extragastrointestinal stromal tumor presenting as a vaginal mass: report of an unusual case with literature review. AB - The clinical and pathologic features of a malignant extragastrointestinal stromal tumor presenting as a vaginal mass are discussed. A 66-year-old female presented with copious vaginal bleeding and spontaneous passage of tumoral tissue per vagina. Histologic assessment showed a mitotically active spindle cell neoplasm. Immunohistochemical analysis demonstrated the neoplasm to be positive for CD117 (transmembrane tyrosine kinase) and CD34, consistent with a malignant extragastrointestinal stromal tumor. Subsequent clinical examination revealed an 8-cm posterior vaginal wall mass, with probable origin from the rectovaginal septum. This case is unique based on the primacy of presenting gynecologic complaints, and the unusual anatomic location of the lesion. A literature review of the pathologic features of extragastrointestinal stromal tumors and factors predictive of biologic behavior are discussed. Correct tumor diagnosis is emphasized given the effective treatment possible with imatinib for patients with unresectable tumors. PMID- 16343206 TI - Bone metastases in vulvar cancer: a rare metastatic pattern. AB - Bone metastases from a vulvar carcinoma are exceptionally rare with only five reported cases in the literature. We report on a patient who was initially treated with radical vulvectomy and bilateral inguinal lymphadenectomy for a vulvar cancer (pT2, pN2 (6/37), M0; G2). Due to a positive nodal status, adjuvant radiation of the vulva and the pelvis was performed additionally. The patient presented 4 months after initial therapy with severe pain in the right humeral shaft due to a pathologic fracture based on an osteoclastic metastasis. During osteosynthetic stabilization histologic and immunohistochemical stain gave evidence of a metastasis of the known vulvar carcinoma. Bone scan showed enhancements in both humeral heads as well as the right distal femur, whereas plain radiographs confirmed further metastases in all suspected areas. In conclusion, bone metastases should be considered in the differential diagnoses of unclear osseous pain in women with a history of vulvar cancer. Immunohistochemical examinations might be important to depict the epithelial character of the tissue and allude to the metastatic nature of such rare lesions. The atypical location should alert the physician to suspect distant metastasis, rather than locoregional disease. PMID- 16343207 TI - Ewing's sarcoma of the vulva--a case report. AB - Extraosseous Ewing's sarcoma of the vulva is extremely rare with only three such cases described. A 26-year-old woman presented with Ewing's sarcoma of the vulva. She received chemotherapy to shrink the lesion. However, the response was suboptimal and she subsequently received radiotherapy. Although resection was planned, she developed chest metastases within a short period of time and subsequently died. PMID- 16343208 TI - Recurrence of high-grade squamous intraepithelial neoplasia in neovagina: case report and review of the literature. AB - Recurrent carcinoma in situ in neovagina is rare, and the optimal modality of treatment is unclear. A 33-year-old multiparous woman was referred for vulvar intraepithelial neoplasia, vaginal intraepithelial neoplasia, and cervical intraepithelial neoplasia, underwent skinning vulvectomy with perianal excision, total vaginectomy, vaginal hysterectomy, and vaginal reconstruction with split thickness skin graft. Ten years after initial surgery, the recurrence as a high grade intraepithelial neoplasia in the upper one third of neovagina was detected. For that reason, the upper one third of vaginectomy with at least 5-mm tumor-free border and vaginal reconstruction with split-thickness skin graft were performed. She has attended her regular follow-up for 3 years with no evidence of disease. All patients with vaginoplasty should undergo regular follow-up. This report is the seventh such report in English literature of patients previously treated for in situ carcinoma who later developed recurrence in the graft. PMID- 16343209 TI - Incisional skin metastasis of a squamous cell cervical carcinoma 3.5 years after radical treatment--a case report. AB - Metastatic carcinoma in an abdominal wall incision from internal malignant neoplasm is an uncommon and often a preterminal event. Most commonly metastatic skin incisional cancers have been reported with cancers of colon, kidney, and bladder. Incisional metastasis from postoperative case of carcinoma cervix is very rare. It has been reported in few patients with adenocarcinoma and poorly differentiated histopathology, more so at drain site postoperatively. We report a postoperative case of squamous cell carcinoma cervix FIGO stage IIA in a patient who after 3.5 years of completion of radical treatment (postoperative external and intravaginal radiation therapy) developed incisional skin metastasis followed by extensive subcutaneous metastasis in the vulval region. She received salvage chemotherapy; however, she did not show any response and finally succumbed to the disease. The outcome of patients with incisional skin metastasis is usually poor as they are often associated with locoregional recurrence. The intent of treatment remains palliation either by radiation/chemotherapy/surgery alone or in combinations. As far as we know, this is the first case of squamous cell carcinoma cervix stage IIA having incisional scar recurrence 3.5 years after postoperative radiotherapy. PMID- 16343210 TI - Villoglandular papillary adenocarcinoma of the uterine cervix responding to neoadjuvant chemotherapy with docetaxel and cisplatin: a case report. AB - Villoglandular papillary adenocarcinoma (VGPA) of the uterine cervix is a rare neoplasm, and its treatment has rarely been reported. We report a patient with VGPA stage IIA responding to neoadjuvant chemotherapy with docetaxel (60 mg/m2 as an intravenous infusion) and cisplatin (70 mg/m2 as an intra-arterial infusion). At 3 weeks after completing one course of this regimen, the tumor size was reduced from 5.3 x 4.0 cm to 2.0 x 2.0 cm (81.1% reduction), revealed by computed tomography. Accordingly, the patient underwent radical hysterectomy, and there have been no signs of recurrence. Thus, the combination of docetaxel and cisplatin is suggested to be useful for neoadjuvant chemotherapy of cervical adenocarcinoma. PMID- 16343211 TI - Simultaneously detected primary malignant tumors of ovary and endometrium with unusual histology. AB - A case of a mucinous adenocarcinoma of the ovary with a synchronous endometroid tumor of the endometrium with focal features of undifferentiated carcinoma and deep myometrial invasion is reported. A review of the literature revealed that our case is interesting in view of the fact that simultaneous presentation of primary ovarian and endometrial neoplasms is rare and usually related to low stage ovarian lesions and well-differentiated and superficial endometrial carcinomas in contrast to our case with the focal features of undifferentiated carcinoma and the deep myometrial invasion. These double tumors usually present in premenopausal subfertile women with abnormal uterine bleeding. The prognosis in most of the cases is surprisingly good even after total abdominal hysterectomy and bilateral oophorectomy alone without adjuvant chemotherapy or irradiation. PMID- 16343212 TI - Endometrial carcinoma with laparotomy wound recurrence: complete remission following surgery and chemotherapy consisting of paclitaxel and carboplatin. AB - We present a patient with surgical stage I endometrial cancer who experienced laparotomy wound recurrence 4 years after primary treatment. She was treated successfully by complete surgical resection of recurrent tumors and chemotherapy. A 62-year-old white female with laparotomy wound recurrence of endometrial carcinoma with small-bowel involvement and concomitant subcutaneous metastasis in the abdominal wall underwent complete surgical resection of metastatic tumors followed by six cycles of chemotherapy consisting of paclitaxel (175 mg/m2) and carboplatin (area under the curve 5). Since 24 months after resection of recurrence, she has no evidence of disease recurrence. Endometrial carcinoma with laparotomy wound recurrences, especially those with concomitant metastases, can be successfully treated by complete surgical resection followed by chemotherapy consisting of paclitaxel and carboplatin. PMID- 16343213 TI - Complete response to docetaxel and carboplatin combination chemotherapy for a stage IV uterine papillary serous carcinoma: a case report. AB - We report a case of a stage IV uterine papillary serous carcinoma (UPSC) with multiple organ metastases. The patient was treated with docetaxel and carboplatin combination chemotherapy. After five courses, uterine tumor, Douglas tumor, lymphadenopathy, and distant metastases on magnetic resonance imaging or computed tomography scan were completely resoluted. Moreover, endometrial biopsy showed no carcinoma tissues after six courses. We suggest that this regimen may be effective for treatment of advanced-stage UPSC. PMID- 16343214 TI - Primary adenocarcinoma arising from endometriosis of the rectovaginal septum: a case report. AB - Adenocarcinomas developing in endometriotic lesions have been reported, most commonly in the ovary. The rectovaginal septum is a rare location of this neoplastic transformation. We report a case of primary adenocarcinoma arising from endometriosis of the rectovaginal septum in a 25-year-old woman presenting with lower abdominal pain and dyspareunia. Diagnostic and treatment modalities were discussed. Immunohistochemical studies of different keratin subtypes and CA125 expression of the tumor cells are of interest for the diagnosis. Preoperative radiation therapy promotes clinical symptoms relief, tumor volume reduction, and better radical surgical approach. PMID- 16343215 TI - Parametrial and rectovaginal adenocarcinoma arising from endometriosis. AB - Malignant extragonadal tumors arising from endometriosis are rare. We report on two cases. A 41-year-old gravida 1, para 1 (G1P1), with adenocarcinoma of the right parametrium arising from endometriosis and a 51-year-old G1P1 with endometriosis-associated rectovaginal adenocarcinoma were treated. Treatment included radical surgery plus radiation therapy. While the former patient was doing well 2 years after the primary diagnosis, the latter suffered a local pelvic recurrence 2 years later. Although there are no randomized controlled studies, radical surgery followed by radiation therapy seems generally to be the treatment of choice. The analysis of PTEN in various forms of endometriosis and its malignant transformation may help in understanding the early steps of tumorigenesis. PMID- 16343216 TI - Report of an unusual problematic uterine smooth muscle neoplasm, emphasizing the prognostic importance of coagulative tumor cell necrosis. AB - Follow-up data of the clinical behavior of uterine smooth muscle tumors with low malignant potential are scarce. We present a woman suffering from a uterine smooth muscle cell tumor with increased cellularity, absence of significant atypia, and two to three mitotic figures per 10 HPFs but with minimal focal coagulative tumor cell necrosis (CTCN). These microscopic features are currently accepted to label the lesion as a "smooth muscle neoplasm of low malignant potential, limited experience." After a disease-free survival of 4 years, two retroperitoneal tumors around the iliac vessels were extirpated. Both tumors consisted of smooth muscle cells with mild to moderate atypia and a mitotic index of 5-10 per 10 HPFs but with multifocal and extensive CTCN. These microscopic features were sufficient to establish the diagnosis of leiomyosarcoma. This case adds to the limited experience of the clinical behavior of problematic uterine smooth muscle cell neoplasms and underscores the prognostic importance of CTCN. PMID- 16343217 TI - Benign metastasizing leiomyoma responsive to megestrol: case report and review of the literature. AB - Benign metastasizing leiomyoma (BML) is an unusual condition typically treated by surgical resection and hormonal therapy. A 37-year-old woman presented with dyspnea 6 years after uterine myomectomy. Computed tomographic scans showed pelvic and right-sided lung masses. Total abdominal hysterectomy and right salpingo-oophorectomy revealed only leiomyomas and benign ovarian cysts. Thoracotomy revealed multiple nodules consistent with leiomyomas. Estrogen and progesterone receptors on lung and uterine tissue were positive. A 21- x 18-mm residual area of neoplasm in the lung completely resolved 3 months after treatment with megestrol. The patient remains free of disease 3 years after lung resection. We present a rare case of BML in which the lung neoplasm responded to megestrol alone in the setting of intact ovarian function. PMID- 16343218 TI - A case of cotyledonoid leiomyoma and review of the literature. AB - We report a case of a rare, unusual benign tumor, the cotyledonoid leiomyoma or Sternberg tumor, detected incidentally in a 67-year-old woman who underwent total hysterectomy and bilateral salpingo-oophorectomy because of a persisting left ovarian cyst of 5 cm. This is the 14th case of cotyledonoid leiomyoma and 2nd case which had no intrauterine portion but had extrauterine extensions. At laparotomy, it was detected that a deep red nodular papillary tumor had been dissecting the posterior leaf of the left broad ligament, projecting into the abdominal cavity just near the left cornu of the uterus, and extending in downward direction. The remaining portion was lying between the layers of the left broad ligament and arose from the subserosa of the left side of the uterine isthmus. Although the bizarre and unusual shape was in favor of malignity, the frozen-section examination revealed a benign histology. Although the cotyledonoid leiomyoma is a benign entity, it may suggest a malignant disease owing to its unusual sarcomatoid appearance and its rarity. Therefore, awareness of obstetricians and gynecologists regarding this rare entity will prevent unnecessary anxiety and interventions. PMID- 16343219 TI - Retroperitoneal leiomyomatosis: a case report. AB - Leiomyoma is the most common benign solid pelvic tumor seen in women. It is most commonly located in the uterus and gastrointestinal tract, but it can originate wherever smooth muscle cells exist. Although it has been reported in various atypical localizations, they are extremely rare in the retroperitoneum. Also, preoperative diagnosis is often difficult in retroperitoneal tumors. Imaging studies may demonstrate the retroperitoneal tumors; however, exact diagnosis cannot be established by imaging methods alone. Here, we report a case with retroperitoneal leiomyomatosis diagnosed by preoperative ultrasonography-guided fine-needle biopsy. PMID- 16343220 TI - Leiomyosarcoma of the broad ligament: a case report and review of literature. AB - Leiomyosarcoma of the broad ligament is a rare tumor, since only 12 cases have been reported so far in the literature. A 53-year-old patient was diagnosed with leiomyosarcoma of the broad ligament at the Department of Gynecology Oncology of the National Cancer Research Institute in Genoa. The tumor had low mitotic activity and less than ten mitotic figures were found for ten high-power fields. The treatment consisted of total abdominal hysterectomy and bilateral salpingo oophorectomy with bilateral ureteral neoanastomosis and omentectomy. The patient has not received either radiotherapy or chemotherapy, considering the low grade of malignancy, but she is only followed up on an outpatient basis. No evidence of metastasis has been noted after a follow-up of 13 months. It is concluded that low-grade leiomyosarcoma of the broad ligament should be treated only with surgery. PMID- 16343221 TI - Uterine liposarcoma in a young woman: a case report. AB - Liposarcoma of uterus is a very rare neoplasm encountered generally in the sixth or seventh decade of life. We reported the eighth case of uterine liposarcoma with extension to broad ligament in a 23-year-old woman who presented with pelvic pain, dysuria, and rapidly enlarging pelvic mass. During laparotomy, the mass was detected to be originated from the uterine cervix. In frozen section, no definitive diagnosis was established, and for preservation of fertility, extirpation of the mass was performed. The histopathologic examination of the specimen revealed liposarcoma. The tumor cells were diffusely and intensively immune reactive for vimentin and S-100, and negative for smooth muscle actin, desmin, h-caldesmon, pan cytokeratin, and CD10. Staining with periodic acid and Schiff, alcian blue, mucicarmine, and epithelial membrane antigen (EMA) were negative. No recurrence was detected in the postoperative seventh month. Present case is the youngest patient with uterine liposarcoma and also the first case detected in reproductive period. PMID- 16343222 TI - Recurrent retroperitoneal myxoid liposarcoma during pregnancy: a case report and literature review. AB - Retroperitoneal liposarcoma in pregnancy is rare, and only a few cases of primary liposarcoma during pregnancy have been reported. To the best of our knowledge, there is no published report of retroperitoneal liposarcoma that was previously treated and recurred during pregnancy. Our patient was diagnosed with a pelvic mass on ultrasound at 12-weeks' gestation. The mass was found to be a retroperitonal, well-differentiated myxoid liposarcoma and was radically excised at the time of cesarean delivery at 36-weeks' gestation. However, the tumor recurred soon and progressed rapidly, and the patient eventually died of the disease. A thorough sonographic investigation and timing of surgery may be critical in terms of finding a surgically resectable lesion and leading to a more favorable prognosis. PMID- 16343223 TI - Development of uterine sarcoma after tamoxifen treatment for breast cancer: report of four cases. AB - Tamoxifen (TAM) is widely used in the treatment of breast cancer, and its paradoxical effects on female genital system are well known. During the past 10 years, many descriptions of nonepithelial uterine malignancies related to long term TAM usage have been reported in the literature. Four uterine sarcoma patients who had history of TAM usage for previous breast cancer are presented in this study. The mean time of exposure to TAM was 6 (range 3-11) years, and the mean cumulative dose of drug was 43.82 g. All patients were postmenopausal, and the mean age was 66 (range 61-73) years at the time of the diagnosis of the uterine malignancy. Two (50%) patients had uterine malignant mixed mullerian tumor, and two (50%) had leiomyosarcoma. In one (25%) patient was diagnosed with endometrial biopsy made for a postmenopausal vaginal bleeding; the others (75%) were asymptomatic and their diseases were diagnosed during the pelvic examination and transvaginal ultrasonography. All patients underwent surgery +/- adjuvant therapy (chemotherapy and/or radiation therapy), and two (50%) patients died because of the sarcoma. In consequence, early detection of TAM-related uterine sarcoma is required for orderly gynecological examination in patients having history of TAM usage for previous breast cancer. PMID- 16343224 TI - Response of recurrent uterine high-grade malignant mixed mullerian tumor to letrozole. AB - Uterine malignant mixed mullerian tumor (MMMT) is a rare malignancy occurring most often in postmenopausal women. Despite the use of multimodality treatments including surgery, chemotherapy, and radiotherapy, prognosis is still poor in most cases. We report the case of a 69-year-old woman with recurrent metastatic high-grade MMMT that responded to letrozole, an aromatase inhibitor. At the initial diagnosis of high-grade uterine MMMT in February 2001, the patient underwent total abdominal hysterectomy, bilateral salpingo-oophorectomy, and postoperative pelvic radiotherapy. Two years later, an asymptomatic retroperitoneal mass was discovered on surveillance abdominal computed tomography scanning. The 3.5- x 3.0-cm mass was considered inoperable owing to its location near the aorta at the level of the renal vessels. The patient declined radiation or chemotherapy. Treatment with letrozole was begun at 2.5 mg daily. Serial computed tomography scans demonstrated marked tumor shrinkage; after 11 months of letrozole therapy, the tumor had shrunk to less than 25% of its original volume. Further study of letrozole for high-grade uterine MMMT is warranted. PMID- 16343225 TI - Malignant mixed mullerian tumor of primary mesenteric origin. AB - Malignant mixed mullerian tumor (MMMT) is a rare tumor. A literature search revealed very few reports on MMMT, especially those arising in the peritoneum. We recently encountered an MMMT of primary mesenteric origin associated with left fallopian tube cancer. There have been no previous reports about its occurrence in the mesentery. When cases of peritoneal MMMT were reviewed, the disease was found to be associated with synchronous or metachronous gynecologic tumors of mullerian duct origin (ie, ovarian tumors, primary serous carcinoma of the peritoneum, fallopian tube cancer, endometrial cancer, and adenocarcinoma of the cervix) in 12 out of 32 patients (37.5%). Peritoneal MMMT are frequently associated with gynecologic tumors. PMID- 16343226 TI - Bariatric surgery in the field of gynecological oncology. AB - Obesity has become a foremost health problem. More than half of US adults are overweight or obese. This has been due to sedentary lifestyles, increased intake of refined carbohydrates, and fat-rich diets. Obese women are particularly susceptible to a variety of health risks including cancer, especially cancers of the breast, endometrium, and colon. Bariatric surgery appears to be a viable option for the treatment of severe obesity. As the role of surgery in the management of this condition becomes increasingly frequent, it is important for gynecological oncologists to recognize the potential for gynecological malignancies in this patient population. PMID- 16343227 TI - Second-look operation improves survival in suboptimal, debulked, stage III ovarian cancer. PMID- 16343230 TI - The role of surgery in recurrent ovarian cancer. AB - The role of cytoreductive surgery (CS) in recurrent ovarian cancer (ROC) has not been clearly defined. We performed a retrospective study evaluating criteria for CS in ROC. Twenty-five institutions documented their patients with CS for invasive epithelial ROC performed 2000-2003. Two hundred sixty-seven patients were included. Complete tumor removal was achieved in 133 patients (50%). Complete resection was associated with prolonged survival compared to surgeries with residual tumor. Median survival of patients without residual tumor was 45.3 months and of patients with residual tumor, irrespective of its size, 19.0 months (HR 4.33; 95% CI 2.53-7.43; P < 0.0001). In a multivariate analysis, the following factors showed a significant influence on the probability to achieve a postoperative residual tumor of 0 mm: absence of ascites (<500 vs > or =500 mL: HR 4.63; 95% CI: 1.81-11.76; P= 0.0001), good performance status Eastern Cooperative Oncology Group (ECOG) 0 vs >0: HR: 2.41; 95% CI: 1.41-4.08; P= 0.001, and low FIGO stage at primary diagnosis (FIGO I/II vs III/IV: HR 1.87; 95% CI: 1.04-3.37; P= 0.036). Significant factors for survival after surgery for recurrence in a multivariate analysis were achievement of complete resection (residual tumor at surgery for recurrence 0 vs >0 mm: HR 2.86; 95% CI: 1.66-4.93; P < 0.001), absence of ascites (<500 vs > or =500 mL: HR 2.09; 95% CI: 1.18-3.71; P= 0.012), and application of a platinum-containing chemotherapy (platinum containing chemotherapy vs others: HR 1.83; 95% CI: 1.16-2.88; P= 0.009). Only patients with complete resection seem to benefit from CS. This new panel of selection criteria will be evaluated in a prospective study. PMID- 16343231 TI - What is the role of conservative primary surgical management of epithelial ovarian cancer: the United States experience and debate. AB - Certification in Gynecologic Oncology and creation of the Society of Gynecologic Oncologists in the United States have led to the development of a specialty with individuals capable of performing complex abdominal and pelvic operations in the management of epithelial ovarian carcinoma. These operations can be divided into two types. 1) A staging operation to assess the extent of disease through careful palpation, histologic and cytologic assessment of all peritoneal surfaces along with removal of the uterus, ovaries and fallopian tubes, omentum, together with a bilateral pelvic and aortic lymphadenectomy. Such information allows the clinician to determine prognosis and if postoperative adjuvant therapy is indicated. 2) A debulking operation designed to resect or reduce the size of metastatic lesions as well as to remove the primary tumor including a bilateral salpingo-oophorectomy. This operation is designed to improve survival and cure. In spite of this apparently clear paradigm, there has been a steady debate as to the apparent justification of these operations, especially when the former is performed in a women who has not completed her childbearing and especially when the latter requires "ultraradical" procedures. Many feel that the pendulum is now swinging toward fertility-sparing surgery among young women with early invasive cancers and toward either neoadjuvant chemotherapy or less than ultraradical debulking among women with advanced ovarian cancer. The purpose of this study is not to provide an exhaustive review but rather to outline this debate and focus on the American experience with conservative surgery in the management of epithelial ovarian carcinoma. PMID- 16343232 TI - Role of conservative surgery in ovarian cancer: the European experience. AB - Although less frequently than in older women, about 15% of invasive epithelial ovarian cancer may occur in young women, for whom preservation of fertility potential is an important clinical goal. We reviewed the published evidences from the European literature on the role of conservative surgery in women with invasive epithelial ovarian cancer. Three reports were identified from the Italian and French literature; the data were analyzed together with our own experience in terms of relapse rate, relapse in the preserved ovary, survival, and fertility outcome. A total of 152 conservative surgeries were reported: 88 patients with stage IA, 2 with stage IB, 51 with stage IC, 2 with stage II, 3 with stage IIIA, and 6 with stage IIIC. Relapses occurred in 18/152 patients (11.8%) and involved the preserved ovary in 11 cases (7%). Fifty-three pregnancies were recorded with 38 uneventful term deliveries, 2 ectopic pregnancies, 6 spontaneous abortions, 4 terminations, and 2 with unknown outcome. Nine patients (5.9%) have died of disease. These findings confirm that young women with stage I invasive epithelial ovarian cancer may receive a successful treatment of their disease without sacrificing fertility. PMID- 16343233 TI - Standard treatment in advanced ovarian cancer in 2005: the state of the art. AB - What are standards? The oncology community expends considerable effort to review the results from definitive treatment studies and define recommendations for future studies, as well as standards of care for the community and patients who are not participating in clinical trials. This is a thoughtful and well intentioned process but subject to considerable bias due to limitations in the data and/or their interpretation. While ovarian cancer is highly responsive to platinum-based therapy after initial cytoreductive surgery, there is a substantial risk of recurrence, which is accompanied by the emergence of drug resistant disease. Better treatments with improved long-term outcomes are needed. From this perspective, standards can help to provide a baseline for assessing gaps in our current knowledge and defining priorities for future clinical trials. While not an exhaustive review, this study will focus on key clinical concepts that are guiding ovarian cancer research and treatment. PMID- 16343237 TI - "Dose dense" chemotherapy in ovarian cancer. AB - In essence, dose densification is "accelerated therapy" (a commonly used phrase in radiotherapeutics) and is a form of dose intensification because the amount of drug per unit time (dose intensity = mg/m(2)/week) is increased. There is general consensus that increasing platinum dose intensity in ovarian carcinoma has not been proven despite a dozen or more randomized trials evaluating up to twofold increases in dose intensity. Few randomized trials in ovarian carcinoma have compared weekly "dose dense" chemotherapy with more conventional dosing schedules although there are plenty of phase II studies. In these, dose densification of single agent therapy, for some drugs at least, appears to be relatively well tolerated, with encouraging levels of activity in patients purportedly refractory to the same agents when scheduled in the standard way. However, many studies ostensibly evaluating "dose density" do not actually evaluate this entity, but actually split the standard 3-weekly dose into weekly fragments thus maintaining the same dose intensity. Furthermore, as the aim of treatment in recurrent ovarian cancer is palliation, weekly treatments are less convenient, are probably less cost effective, and have different dose-limiting toxicities. This article will review the clinical data supporting dose density as a therapeutic maneuver in ovarian cancer. PMID- 16343238 TI - What is the role of dose-dense therapy? AB - The introduction of paclitaxel/platinum combination chemotherapy and (interval) debulking surgery has significantly improved the prognosis of patients with ovarian cancer. Yet, many patients die of drug-resistant disease. Second-line chemotherapy may result in prolonged secondary remissions with alleviation of symptoms and improvement of quality of life. The response to second-line chemotherapy is strongly related to platinum sensitivity. More than 60% of platinum-sensitive patients respond to a re-challenge with platinum-containing chemotherapy. In platinum-resistant patients, on the contrary, the response rate to a re-challenge with 3-weekly platinum or any nonplatinum chemotherapy is less than 20%. The response to dose-dense weekly platinum-based regimens ranged from 48% to 64% in platinum-resistant patients. Moreover, the majority of the patients responded within 8 weeks after the start of the treatment. The progression-free survival ranged from a median of 5 months in a study using cisplatin/etoposide, to 11 months in a study with paclitaxel/carboplatin. The median survival was 11 15 months. The outpatient weekly paclitaxel/carboplatin regimen, with paclitaxel at a dose of 90 mg/m(2) and carboplatin at area under the curve 4, seems similarly effective and is better tolerated. Dose-dense weekly paclitaxel/carboplatin is an effective and well-tolerated therapy for platinum sensitive, as well as platinum-resistant tumors. Responses to therapy are observed within 8 weeks in the majority of the patients. Whether a weekly regimen indeed is more effective than 3-weekly paclitaxel/carboplatin needs to be answered in a randomized study. PMID- 16343239 TI - Treatment of recurrent disease: randomized trials of monotherapy versus combination chemotherapy. AB - Recurrent ovarian carcinoma is normally an incurable disease situation in which chemotherapy is the usual treatment for palliation. The probability of response to chemotherapy depends on the time from last chemotherapy and the previous response observed. The issue of combination chemotherapy versus monotherapy is a clinically relevant dilemma for medical and gynecologic oncologist involved in the treatment of recurrent patients. In those patients with platinum-resistant relapse, combination chemotherapy has been associated with higher toxicity without a clear clinical benefit in randomized clinical trials. Therefore, a less toxic sequential monotherapy approach should be the choice for resistant patients. On the other hand, two large randomized clinical trials have proved the superiority of a platinum-based doublet over platinum monotherapy in platinum sensitive recurrent patients. The ICON-4/AGO-OVAR 2.2 trial demonstrated that the combination of paclitaxel-carboplatin (or cisplatin) is likely to provide a survival benefit compared with carboplatin monotherapy. This benefit was more clear in patients with a treatment free-interval >12 months. Moreover, the AGO OVAR 2.5 trial, with the cooperation of NCIC CTG and EORTC GCG, has confirmed the advantage in response rate and progression free survival of the doublet carboplatin-gemcitabine compared to carboplatin. PMID- 16343240 TI - Randomized single-agents trials in recurrent epithelial ovarian cancer. AB - The response to salvage treatment in recurrent epithelial ovarian cancer (REOC), is influenced by many biologic features which should be taken into account in the process of therapeutic decision. Until recently, single agents have been considered effective as combination chemotherapy in REOC and they still represent an option for well-defined categories of patients. In clinical practice, the selection of drugs for second-line treatment can be based on the knowledge that tumor size and response to prior platinum are predictors of response and that the efficacy of some commonly used single agents is supported by well designed clinical studies. Only two single-agent randomized studies with long-term survival analyses have been published; in the topotecan versus paclitaxel study, performed on a total of 235 patients, the long-term survival results did not confirm the initial report of a superiority of topotecan, with a median survival of 63 weeks and 53 weeks for patients treated with topotecan, and, respectively, paclitaxel. No comparative data were provided in the subgroup of potentially platinum-sensitive disease. In the phase III study of pegylated liposomal doxorubicin versus topotecan, into which 474 patients were treated between 1997 and 1999, the higher efficacy of pegylated liposomal doxorubicin was even more evident in the long-term survival analysis, with a 18% reduction in the risk of death, in the overall population which increased to 30% in the subset of platinum sensitive patients. The survival after the two treatments, however, was not different in the group of patients with platinum-refractory disease. The toxicity profile of the two drugs was completely different, with palmar-plantar erythrodysestesia as most common adverse event after pegylated doxo (35% of patients) and severe neutropenia after topotecan (77% of patients). These data confirm the role of pegylated liposomal doxorubicin in the management of REOC and indicate that the identification of new active drugs in this disease is relevant but also feasible only in platinum-sensitive patients. PMID- 16343241 TI - New agents for the treatment of ovarian cancer: the next generation. AB - Ovarian cancer shares many important characteristics with more common malignancies including breast, lung, and colon cancer. The relative chemosensitivity of ovarian cancer and other aspects of its unique biology provide opportunities for novel interventions. In this brief summary, some of the potential targets in ovarian cancer are discussed, including the HER kinases, heat shock protein, the 26S proteasome, and the angiogenesis pathway. The opportunities to change the treatment of ovarian cancer will require creative clinical trial design but the next decade promises to be filled the therapeutic advances for patients with ovarian cancer. PMID- 16343242 TI - Trial design for testing new therapies in ovarian cancer. PMID- 16343243 TI - Ovarian cancer in the proteomics era: diagnosis, prognosis, and therapeutics targets. AB - Ovarian cancer is clinically quiet as it plants seeds of metastases in the peritoneal cavity, even during early stages when there is highest potential for cure. The only available biomarker is CA125, which has an unacceptably low sensitivity and specificity for diagnostic use. Highly sensitive and specific tools to further optimize early diagnosis and treatment are needed. We propose that proteomic technologies have an important role to play in the development of these tools. Mass spectrometry platforms, such as surface-enhanced laser desorption/ionization time-of-flight, may be used to mine patient's serum for proteomic signatures that are shed by tumor and stroma. Such signatures could serve as a diagnostic tool during early-stage disease and as a remission monitoring tool in later-stage disease. Reverse-phase protein microarrays are a new microproteomic tool to profile signaling pathways in ovarian cancer, thus identifying therapeutic targets while simultaneously suggesting prognostic indicators. Proteomic technologies have the capacity to build upon our genomic and clinical understanding of ovarian cancer by moving the focal point to the tumor and its microenvironment. This unique proteomic vantage point allows the creation of tools that will aid clinicians in making rational decisions in the diagnosis and treatment of women with ovarian cancer. PMID- 16343244 TI - New tumor markers: CA125 and beyond. AB - A variety of biomarkers have been developed to monitor growth of ovarian cancer and to detect disease at an early interval. CA125 (MUC16) has provided a useful serum tumor marker for monitoring response to chemotherapy, detecting disease recurrence, distinguishing malignant from benign pelvic masses, and potentially improving clinical trial design. A rapid fall in CA125 during chemotherapy predicts a favorable prognosis and could be used to redistribute patients on multiarmed randomized clinical trials. Several studies now document that CA125 can serve as a surrogate marker for response in phase II trials. Serial measurement of CA125 might also provide a useful marker for monitoring stabilization of disease with cytostatic targeted therapeutic agents. The greatest potential for serum markers may be in detecting ovarian cancer at an early stage. A rising CA125 can be used to trigger transvaginal sonography (TVS) in a small fraction of patients. An algorithm has been developed that calculates risk of ovarian cancer based on serial CA125 values and refers patients at highest risk for TVS. Use of the algorithm is currently being evaluated in a trial with 200,000 women in the UK that will test critically the ability of a two stage screening strategy to improve survival in ovarian cancer. Whatever the outcome, as 20% of ovarian cancers have little or no expression of CA125, additional serum markers will be required to detect all patients in an initial phase of screening. More than 30 serum markers have been evaluated alone and in combination with CA125 by different investigators. Some of the most promising include: HE4, mesothelin, M-CSF, osteopontin, kallikrein(s), and soluble EGF receptor. Two proteomic approaches have been used: one examines the pattern of peaks on mass spectroscopy and the other uses proteomic analysis to identify a limited number of critical markers that can be assayed by more conventional methods. Both approaches are promising and require further development. Several groups are placing markers on multiplex platforms to permit simultaneous assay of multiple markers with very small volumes of serum. Mathematical techniques are being developed to analyze combinations of marker levels to improve sensitivity and specificity. In the future, serum markers should improve the sensitivity of detecting recurrent disease as well as facilitate earlier detection of ovarian cancer. PMID- 16343245 TI - Interleukin-10 and Fas polymorphisms and susceptibility for (pre)neoplastic cervical disease. AB - Infection with oncogenic types of human papillomavirus (HPV) is the main causal factor of cervical cancer and its precursor lesion (cervical intraepithelial neoplasia [CIN]). Cellular immunity may be critical in the elimination of HPV harboring cells. Interleukin-10, a T-helper type 2 cytokine, has a suppressive effect on cell-mediated immunity. Resistance to apoptosis through the Fas pathway might enable many cancers to escape the immune system. We examined in a large study population whether three polymorphisms in the IL-10 gene and a polymorphism at position -670 of the Fas promotor affect susceptibility for cervical cancer or its precursor. In addition, it was studied whether these polymorphisms were causal and not merely associated by typing microsatellite markers in the region surrounding both genes. A total of 311 CIN, 695 cervical cancer patients, and 115 family-based and 586 unrelated controls were analyzed. Association analysis revealed an increased CIN (II-III) (OR 1.44 [1.06-1.97]) and squamous cell carcinoma of the cervix (OR 1.35 [1.04-1.75]) for individuals heterozygous for the A-allele of the IL-10-592 polymorphism. In contrast to previous findings, no association was found for the IL-10-1082 polymorphism. While an increased risk for adenocarcinoma (AC) in heterozygotes (OR 1.59 [1.02-2.48]) was observed. Our study shows a possible role for the IL-10 gene in CIN and squamous cell cervical cancer susceptibility in the Caucasian population; simultaneously, there might be a role for the Fas gene in the development of AC of the cervix. Further investigations with a higher density of markers are necessary to find the causal mutation. PMID- 16343246 TI - Is there still a role for hormonal therapy? AB - Ovarian cancer arises from the specialized surface epithelium, which routinely expresses receptors for reproductive hormones and growth factors. However, as a group, ovarian tumors are less responsive to hormonal therapy than either breast or endometrial cancer. The complex factors that govern hormonal response and associated molecular interactions are under intensive investigation, with the generation of new hypotheses that merit clinical evaluation. As such, it is reasonable to consider a number of potential hormonal interventions related to the prevention and treatment of ovarian cancer. PMID- 16343247 TI - Ovarian cancer: is the news good enough? AB - Ovarian cancer (OC) is still the fourth cause of death by cancer among women and the most fatal among gynecological tumors. The purpose of the symposium held every 2 years in Valencia is to report and discuss new developments in the treatment of OC patients. It also tries to clarify what is evidence-based and what is not. In this issue, proteomic advances for early diagnosis and new tumor markers are presented. Controversies about the role of conservative surgery or the role of surgery in the recurrent disease are discussed. In addition, the current standard treatment, new tendencies on therapy, as well as important aspects related to the molecular era have been updated. Reaching international consensus is a challenge but offers the opportunity to test multiple regimens more efficiently against a single control population, rather than conducting multiple smaller studies. Most of the studies presented in this Symposium were carried out in Gynecologic Cancer Intergroup. If indeed answers to the relevant questions are to be obtained more quickly, then a network of current national or international groups could potentially facilitate this. PMID- 16343248 TI - Fetal signals and parturition. PMID- 16343249 TI - Detection of fetal cells from transcervical mucus plug before first-trimester termination of pregnancy by cytokeratin-7 immunohistochemistry. AB - AIM: The presence of fetal cells in the endocervical mucus of pregnant women was first reported in 1971. The uterine cavity is patent during the first trimester prior to fusion of amnion and chorion. Fetal cells from degenerating chorion frondosum are theoretically shed into the uterine cavity between seven and 13 weeks' gestation and are trapped in the transcervical mucus; they can be identified by immunohistochemistry. METHOD: Ninety-nine transcervical mucosal plugs from pregnant women of between 7 and 13 weeks before abortion were collected, fixed, embedded, sectioned and stained with monoclonal antibody of cytokeratin-7 (CK-7) by immunohistochemistry. RESULT: The identification of trophoblasts on each slide was defined according to positive staining and histologically chorionic villous similarity under microscopic examination, using the following five categories: none (1), less than five single positive-stained cells per-section (2), more than five single positive-stained cells per-section (3), clumps of positive-stained cells (4), and histological-like intact or fragmented chorionic villi (5). From amongst 71 samples that qualified for analysis, individual slides were evaluated and categorized into three groups, with the following results: 32 (45.1%) fell into group 1 (category 1) denoting a negative result, 10 (14.1%) fell into group 2 (category 2) indicating a possible positive result and 29 (40.8%) fell into group 3 (any combination of categories 3 5) representing a positive result. CONCLUSIONS: Fetal cells, identified by CK-7, can be found in more than 59.2% of the transcervical mucus in early pregnancy by use of a minimally invasive sampling method. Prenatal diagnosis of single-gene or chromosome disorders may be possible in the pregnant transcervical mucus by use of modern molecular methods and they deserve further study. PMID- 16343250 TI - Is cytogenetic diagnosis of 46,XX karyotype spontaneous abortion specimens erroneous? Fluorescence in situ hybridization as a confirmatory technique. AB - AIM: Performing the standard cytogenetic technique on spontaneous abortion material is still a valuable tool, but finding a normal 46,XX karyotype can confuse investigators and lead to a problem in diagnosis. This is mainly because it is possible for the female or male conceptus to retain contaminating maternal cells. To address this possibility, we used fluorescence in situ hybridization technique (FISH). X (DXZ1: p11.1-q11.1 region) and Y (DYZ3: p11.1-q11.1 region) chromosome alpha-satellite probes were employed to confirm the karyotypes previously diagnosed as 46,XX by our cytogenetic laboratory, or to verify the occurrence of 'Y chromosome component'. METHODS: Besides conventional long-term tissue cultures and G-bands by trypsin using Giemsa (GTG) bandings, FISH analyses were also performed. RESULTS: A total of 134 spontaneous abortion specimens (singleton gestations) were referred for cytogenetic evaluation, of which 125 specimens were successfully karyotyped. Of these, 20.8% (26/125) had chromosome aberrations; 88.5% (23/26) of these aberrations were numerical and 11.5% (3/26) were structural. The most prevalent numerical anomalies were trisomies 15, 16 and 21, tetraploidies, triploidies and monosomy X. FISH results were obtained for 45 out of 92 cases with 46,XX, of which 2 (4.4%) showed XY signals. CONCLUSIONS: For accurate cytogenetic evaluation of spontaneous abortion materials, an additional technique such as FISH is required in order to confirm the cytogenetic results or to provide an estimate of the error rate in the analysis of miscarriages. PMID- 16343251 TI - Late recurrence of invasive cervical cancer: twenty years' experience in a single cancer institute. AB - AIM: To estimate the characteristics of, and prognosis for, late recurrence of cervical cancer. METHOD: Between 1977 and 1996, of 1721 patients with invasive cervical cancer, 1157 survived more than 5 years. Twenty-eight patients whose cancer recurred after a disease-free interval of 5 years were retrospectively investigated. For 22 patients the cancer recurred after less than 10 years, for five patients it recurred between 10 and 20 years after the original occurrence, and in one patient the cancer recurred after more than 20 years. RESULTS: The histologic subtypes included 20 squamous cell carcinomas and eight adenocarcinomas. The initial therapies were surgery alone (on one patient), surgery and adjuvant radiotherapy (on 20 of the patients) and radiotherapy alone (on seven of the patients). Recurrence was initially diagnosed by internal or external examination (11 patients), chest X-ray (six patients) and computed tomography (11 patients). Rates of lymphogenous metastases were 19% of patients with primary surgery and 57% of patients without primary surgery, while those of hematogenous metastases were 48% of patients with primary surgery and none of patients without primary surgery (P = 0.023). The overall survival rate after recurrence was 50.5% after 2 years and 22.3% after 5 years, respectively. Four of the patients survived for more than 3 years without evidence of disease; these patients all had recurrence outside the irradiated field and achieved complete remission with radiotherapy. CONCLUSIONS: Late recurrence of cervical cancer was frequently observed in patients with initial treatment, including radiotherapy. Some had very long disease-free intervals, but many were diagnosed by ordinary internal or external examination or chest X-ray. Patients with primary surgery experienced recurrence less frequently in the case of lymph node metastasis than was the case for hematogenous metastasis. Radiotherapy was effective and improved prognosis, especially for recurrence of cancer outside the irradiated field. PMID- 16343252 TI - Two cases of reversible posterior leukoencephalopathy syndrome, one with and the other without pre-eclampsia. AB - Two cases of reversible posterior leukoencephalopathy syndrome (RPLS) are reported. One was a 26-year-old woman, who had pre-eclampsia and developed cortical blindness and subsequent eclampsia at 28 weeks' gestation. The other was a 27-year-old woman, who had no pre-eclampsia and developed loss of consciousness and subsequent systemic convulsion at 36 weeks' gestation. On brain magnetic resonance imaging, they both had high signal intensity on T2-weighted and fluid attenuated inversion recovery images, and normal signal intensity on diffusion weighted image of the posterior lobe, which almost disappeared with the amelioration of clinical symptoms thereafter. RPLS is considered to be the result of vasogenic brain edema caused by hypertension. Two hypotheses are conceived to explain the emergence of RPLS without hypertension. The first suggests that an immunotolerant condition such as pregnancy can easily cause vasogenic edema without the elevation of blood pressure. The second suggests that hypertension exists but cannot be detected because it is extremely acute and transient. PMID- 16343253 TI - Spontaneous uterine rupture at 35 weeks' gestation, 3 years after laparoscopic myomectomy, without signs of fetal distress. AB - Laparoscopic myomectomy (LM) is a recently developed surgical technique, and every obstetrician should be aware of its possible complications, which can occur not only during labor but also during pregnancy. We report a case of a primigravid woman who was hospitalized at 35 weeks' gestation because of irregular abdominal pain. She conceived spontaneously 3 years after LM. After a 20-h stay on the obstetrician ward due to increased abdominal tenderness and vaginal bleeding, the patient was qualified for an emergency cesarean section without any symptoms of fetal distress in cardiotocography. During cesarean section a newborn with 9 Apgar points was delivered and a rupture of the uterine wall was seen. Dehiscence of the pregnant uterus following LM is an incidental case, and can therefore be misdiagnosed. Close attention should be paid to every pregnancy in previously operated uteri as the dehiscence of the pregnant uterus can occur without symptoms of fetal distress. PMID- 16343254 TI - Metastasis of uterine leiomyosarcoma to the pancreas. AB - BACKGROUND: Metastasis of uterine leiomyosarcoma to the pancreas is extremely rare. CASE: A 46-year-old woman presented with hypermenorrhea and dysmenorrhea and underwent surgery. The histologic and immunohistochemical diagnosis was uterine leiomyosarcoma stage I with no metastasis to the ovaries or the pelvic lymph nodes. The mitotic count was very high. Thereafter, recurrences in the lung and subsequently in the pancreas were detected. The lesions in the lung and pancreas were resected and diagnosed as metastases of uterine leiomyosarcoma based on histology and immunohistochemistry. CONCLUSION: We report an extraordinarily rare case of uterine leiomyosarcoma with metastasis to the pancreas following initial metastasis to the lung, both of which were diagnosed using histology and immunohistochemistry of the specimens obtained at surgery. PMID- 16343255 TI - Left atrial mechanical functions in pre-eclampsia. AB - AIM: To assess the effect of short-term pressure overload on left atrial (LA) mechanical function in pre-eclampsia. METHODS: Twenty women with pre-eclampsia and 17 age-matched healthy pregnant women were included. LA volumes were measured echocardiographically at the time of mitral valve opening (Vmax), onset of atrial systole (p-wave at the electrocardiography = Vp) and mitral valve closure (Vmin) according to the biplane area-length method. RESULTS: The mean age, gestational age, weight and body surface area were similar in pre-eclampsia and controls. The ventricular septal and posterior wall thickness were greater in pre-eclampsia (P < 0.001). There were no significant differences in LA diameter, Vmax, Vmin, Vp, LA-passive emptying volume, LA-passive emptying fraction, LA-active emptying volume, LA-active emptying fraction, conduit volume, LA-total emptying volume and LA-total emptying fraction between the groups. CONCLUSION: Left atrial mechanical function didn't change in pre-eclampsia. We conclude that short-lasting pressure overload is not capable of inducing changes in LA function. PMID- 16343256 TI - Heterogeneity in alpha-thalassemia interactions in Malays, Chinese and Indians in Malaysia. AB - AIM: Interactions between different determinants of alpha-thalassemia raises considerable problems, particularly during pregnancies where antenatal diagnosis is necessary. This study aims to determine the different types of deletional alpha-thalassemia and Hemoglobin Constant Spring (HbCS), and their frequency in Malays, Chinese and Indians in Malaysia. METHODS: DNA from 650 pregnant women from the Antenatal Clinic of the University of Malaya Medical Center in Kuala Lumpur, Malaysia who showed mean cell volume < or =89 fL and/or mean cell hemoglobin < or =28 pg were analyzed for the double alpha-globin gene South-East Asian deletion (--SEA), the -alpha3.7 and -alpha4.2 single alpha-globin gene deletions and HbCS. RESULTS: One hundred and three (15.8%) of the pregnant women were confirmed as alpha-thalassemia carriers: 25 (3.8%) were alpha-thalassemia-1 carriers with the --SEA/alphaalpha genotype, 64 (9.8%) were heterozygous for the alpha3.7 rightward deletion (-alpha3.7/alphaalpha), four (0.6%) were heterozygous for the -alpha4.2 leftward deletion (-alpha4.2/alphaalpha), nine (1.4%) were heterozygous for HbCS (alphaCSalpha/alphaalpha) and one (0.2%) was compound heterozygous with the -alpha3.7/alphaCSalpha genotype. The double alpha-globin gene --SEA deletion was significantly higher in the Chinese (15%) compared to the Malays (2.5%) and not detected in the Indians studied. The -alpha3.7 deletion was distributed equally in the three races. HbCS and -alpha4.2 was observed only in the Malays. CONCLUSION: The data obtained gives a better understanding of the interactions of the different alpha-thalassemia determinants in the different ethnic groups, thus enabling more rapid and specific confirmation of alpha thalassemia in affected pregnancies where antenatal diagnosis is necessary. PMID- 16343257 TI - Predicting fetal anemia by using reference centile charts for liver length, spleen perimeter and umbilical vein maximum flow velocity in Thai fetuses throughout gestation. AB - AIM: To create reference centile charts for liver length, spleen perimeter and umbilical vein maximum flow velocity (UVVmax) in Thai fetuses in order to predict fetal anemia in Thai fetuses. METHODS: The study was a prospective, cross sectional study, carried out at the Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Faculty of Medicine, Siriraj Hospital, Mahidol University. A total of 780 pregnant women between 13 and 40 weeks' gestation, who attended the antenatal clinic at Siriraj Hospital, Mahidol University, Bangkok, were recruited. Each fetus was measured only once for the purpose of this study. The mean and standard deviation (SD) were estimated at each week of gestation using linear regression modeling. A total of 752 fetuses were measured for fetal liver length, spleen perimeter and UVVmax. Linear regression models were fitted to estimate the mean 95% confidence interval for liver length, spleen perimeter and UVVmax at each gestational age. The centile charts of those parameters were also presented. CONCLUSION: Reference centile charts for fetal liver length, spleen perimeter and UVVmax of Thai fetuses were created. PMID- 16343258 TI - Fetal hydrops associated with congenital pulmonary myofibroblastic tumor. AB - Abstract We report on a fetus with a congenital pulmonary myofibroblastic tumor, the prenatal detection of which with imaging modalities has not been reported up until now. A 32-year-old woman was referred to our hospital at 29 weeks' gestation because of severe fetal hydrops. Sonograms and magnetic resonance imaging showed a large solid tumor in the left thorax. The fetus died in utero the next day. Autopsy confirmed that the tumor was confined to the lower lobe of the left lung, and circulatory insufficiency from compression by the tumor was considered to be the cause of fetal hydrops and demise. Histologic examination revealed that the tumor was composed of uniform short spindle cells with no atypia and a large number of vessels. In addition, with immunohistochemical studies, the tumor cells were stained for calponin but not for cluster differentiation (CD)-31, CD-34, alpha-smooth muscle actin or S-100. PMID- 16343259 TI - Surgical indications for combined partial rectosigmoidectomy in ovarian cancer. AB - AIM: To evaluate surgical indications for combined partial rectosigmoidectomy in ovarian cancer with direct invasion of the rectum and sigmoid colon or dissemination into the pouch of Douglas. METHODS: Subjects comprised 25 patients with ovarian cancer who underwent primary surgery and rectosigmoidectomy between 1990 and 2002 at our hospital. Federation of Obstetrics and Gynecology staging of tumors was II (n = 6), III (n = 17) or IV (n = 2). The histologic type was serous adenocarcinoma (n = 18), clear cell adenocarcinoma (n = 4), and others (n = 3). Bowel resection was performed during primary surgery in 18 patients, and after neoadjuvant chemotherapy (NAC) in seven patients. Cumulative survival rate was compared between NAC and non-NAC groups. Patients were divided into three groups based on extent of surgical resection to compare survival rates: no residual tumor (n = 19); maximum residual tumor diameter <1 cm (n = 5); and maximum residual tumor diameter > or =1 cm (n = 1). RESULTS: Cumulative 5-year survival was 41.3% for all patients. Cumulative 5-year survival in the 18 patients who underwent bowel resection during primary surgery was 62.2%, compared to 13.9% in the seven patients who underwent bowel resection after NAC. Cumulative 5-year survival based on extent of surgical resection was: no residual tumor, 60.8%; residual <1 cm, 0%; and residual > or =1 cm, 0%. Cumulative 5-year survival for patients with complete tumor resection (no residual tumor), excluding clear cell adenocarcinoma, was 79.5%. CONCLUSION: In ovarian cancer with direct invasion of the rectum or sigmoid colon or dissemination into the pouch of Douglas, complete tumor resection with rectosigmoidectomy during primary surgery is associated with good clinical outcomes. PMID- 16343260 TI - Occurrence of fetal choroid plexus cysts in siblings: concerns regarding recurrence and chromosomal abnormality. AB - Choroid plexus cysts (CPC) are a well-known ultrasound aneuploidy marker easily detectable at second-trimester ultrasound examination. However, their genetic etiology is totally unknown. We report two cases of Japanese mothers who carried two and three siblings respectively; all the fetuses that had CPC were noticed at second trimester. Genetic amniocentesis revealed that each fetus had different karyotypes, that is, trisomy 18 and 46,XX in the case of one mother, and trisomy 18, 46,XY and trisomy 21 in the case of the other. These observations indicate that the genetic basis of the cysts is not linked to abnormal chromosomes. We propose that careful ultrasound observation and genetic counseling of the siblings should be offered to patients who have previously had a baby with CPC, despite that baby having a normal karyotype. PMID- 16343261 TI - Microwave endometrial ablation for menorrhagia caused by large submucous myomas. AB - AIM: To examine the feasibility of microwave endometrial ablation (MEA), using a curved microwave applicator, in patients with menorrhagia caused by a submucous myoma greater than 3 cm in diameter, which is among the contraindications for conventional endometrial ablation. METHODS: Patients included were refractory to medication, more than 45 years of age, and due to undergo hysterectomy to treat menorrhagia as a result of submucous myomas, although they hoped to avoid hysterectomy. Three patients underwent MEA at 2.45 GHz using a curved microwave applicator and microwave tissue coagulator to treat menorrhagia. Patients 1, 2 and 3 had submucous myoma nodes 4.5, 5.0 and 13 cm in diameter respectively. The applicator was guided under transabdominal ultrasonography to microwave irradiation sites arranged to cover the entire uterine lining based on preoperative magnetic resonance images. MEA was performed under general anesthesia or spinal anesthesia. Microwaves were irradiated at 40 W for 50 s for single irradiation sites. RESULTS: Patient 1 became amenorrheic after the second MEA, which was performed 2 months after the first operation. Patients 2 and 3 became amenorrheic after the first MEA. Postoperative abdominal pain was controllable by a suppository of non-steroidal anti-inflammatory drugs. The patients were discharged the day after the operation. No remarkable complications were encountered during or after surgery. CONCLUSION: Microwave endometrial ablation improves menorrhagia caused by large submucous myomas when the microwave applicator reaches all parts of the uterine cavity. PMID- 16343262 TI - Discordance in the decline in regional lean and bone mass with advancing age. AB - AIM: To investigate whether regional mineral-free lean mass (lean mass) and bone mineral density (BMD) decrease equally with advancing age. METHODS: Subjects were 420 premenopausal women and 239 postmenopausal women with right-side dominance. Age, height, weight, and body mass index were recorded. Lean mass of the arms, trunk and legs were measured by dual-energy X-ray absorptiometry (DEXA). BMD of the same segmental regions was measured by DEXA. RESULTS: Trunk and bilateral leg lean mass were inversely correlated to age (r = -0.240, P < 0.0001, -0.167, P < 0.0001, and -0.183, P < 0.0001, respectively), but arm lean mass did not decrease with aging. Regional lean mass was positively correlated to regional BMD (r = 0.284-0.449, P < 0.0001). BMD was inversely correlated to age in all segmental regions (r = -0.586 to -0.449, P < 0.0001). CONCLUSIONS: Decline in regional lean mass and decline in BMD with advancing age are discordant. Arm lean mass may not contribute to 'age-related decline' in arm BMD, because arm lean mass does not decrease with advancing age in the general population of women. PMID- 16343263 TI - Case of a giant mucinous ovarian carcinoma with bone metastasis. AB - We report the case of a 51-year-old Japanese woman with a giant (50.75-kg) ovarian tumor. The histopathologic diagnosis was mucinous cystadenocarcinoma. After surgery, the patient was intubated and connected to a respirator for 8 days. Thereafter, she was diagnosed with bone metastasis to the hip bone and the femur. PMID- 16343264 TI - Routine histopathologic analysis of product of conception following first trimester spontaneous miscarriages. AB - AIM: To evaluate the histopathologic findings relating to tissue samples collected at surgical uterine evacuation in first-trimester spontaneous miscarriages. METHODS: In this retrospective study, histopathologic diagnosis of the tissue samples obtained via surgical uterine evacuation in patients who were admitted to the Early Pregnancy Clinic in a 12-month period with the diagnosis of incomplete miscarriage (n = 970), missed miscarriage (n = 406) and anembryonic miscarriage (n = 230) in the first trimester was recorded and compared with the presurgery diagnosis. RESULTS: Uterine evacuation was performed in cases of incomplete miscarriage (n = 970, 60.4%), missed miscarriage (n = 406, 25.2%) and anembryonic miscarriage (n = 230, 14.3%). Histopathologic examination revealed the product of conception in 1119 patients (69.7%), while partial hydatidiform mole was diagnosed in 33 patients (2.1%). Complete hydatidiform mole was detected in only seven cases (0.43%). Exaggerated placental site and placental site trophoblastic nodule was detected in two cases (0.12%). Decidual tissue without chorionic villi was reported in 272 patients (16.9%), raising the suspicion of presence of other pathology. CONCLUSIONS: By routine histopathologic assessment of products of first-trimester spontaneous miscarriages, important pathologies such as molar pregnancy and placental trophoblastic disease can be diagnosed. Histopathological assessment has great value in the identification of an ectopic pregnancy or infection when compared with clinical and laboratory findings. PMID- 16343265 TI - Transplantation of umbilical cord blood-derived endothelial progenitor cells: a promising method of therapeutic revascularisation. AB - Therapeutic neovascularisation by endothelial progenitor cells (EPCs) mediated vascular regeneration is becoming a novel option for the treatment of ischaemic diseases. Recently, human umbilical cord blood (CB) has been found to contain a large number of EPCs and transplantation of CB EPCs led to a successful salvage of the ischaemic limbs through improvement in blood perfusion, indicating the feasibility of using CB cells for therapeutic revascularisation. This review will summarise recent studies in therapeutic revascularisation using CB cells and discuss the potential clinical utilisation of CB cells in ischaemic diseases. PMID- 16343266 TI - Allogeneic haematopoietic cell transplantation for chronic myelogenous leukaemia in the era of imatinib: a retrospective multicentre study. AB - OBJECTIVE: To analyse the results of allogeneic haematopoietic cell transplantation (HCT) in patients with advanced stages of Philadelphia chromosome positive chronic myelogenous leukaemia (CML) who had previously been treated with imatinib mesylate (IM). METHODS: We analysed the outcome of 61 patients with CML who had received allogeneic HCT from sibling (n = 18) or unrelated (n = 43) donors after having been treated with IM. Forty-one patients had received IM because of accelerated or blast phase CML. Conditioning therapy contained standard doses of busulfan (n = 25) or total-body irradiation (n = 20) in conjunction with cyclophosphamide in the majority of cases. Sixteen patients received dose-reduced conditioning with fludarabine-based regimens. RESULTS: The incidence of grades II-IV and III-IV graft-versus-host disease was 66% and 38% respectively. The probability of overall survival (OS), disease-free survival (DFS) and relapse at 18 months for the whole patient cohort were 37%, 33% and 24% respectively. The probability of non-relapse mortality (NRM) at 100 d and 12 months was 30% and 46% respectively. Univariate analysis showed that fludarabine based conditioning therapy, age > or = 40 yr and >12 months interval between diagnosis and transplantation were associated with a significantly lower OS and DFS and a higher NRM. CONCLUSION: These data suggest that although pretreatment with IM is not an independent negative prognostic factor, it cannot improve the dismal prognosis of CML patients at high risk for transplant-related mortality. PMID- 16343267 TI - Molecular analysis of PDGFRalpha/beta genes in core binding factor leukemia with eosinophilia. AB - Eosinophilia sometimes occurs in acute myeloid leukemia (AML), especially in core binding factor (CBF) leukemia. However, the pathogenesis of the differentiation from leukemic progenitors to eosinophils is not well understood in this type of leukemia. Recent reports showed that a novel fusion tyrosine kinase, Fip1-like1 (FIP1L1) platelet-derived growth factor receptor alpha (PDGFRalpha), is found in idiopathic hypereosinophilic syndrome. The involvement of another chimeric gene, PDGFRbeta, was also reported in myeloproliferative disorder with eosinophilia. These chimeric genes cause constitutive activation of PDGFR tyrosine kinases. On the other hand, a two-hit model for the pathogenesis of AML, which seems to be caused by inactivating mutations in transcription factors and genetic lesions in tyrosine kinase resulting in constitutive activation, has been proposed. On the basis of these findings, we screened for the expression of the FIP1L1-PDGFRalpha fusion gene and for mutations in the juxtamembrane and tyrosine kinase domains of PDGFRalpha/beta genes in 22 cases of CBF leukemia with eosinophilia. Among these cases, no FIP1L1-PDGFRalpha fusion gene was found. Although cDNA sequencing also detected three types of single-nucleotide alterations at kinase domains in PDGFRalpha/beta genes, all of them were silent changes and polymorphisms. Therefore, PDGFRalpha/beta genes do not appear to play a significant pathogenetic role in eosinophilia or leukemogenesis of CBF leukemia. PMID- 16343268 TI - Promoter hypermethylation of p15INK4B, HIC1, CDH1, and ER is frequent in myelodysplastic syndrome and predicts poor prognosis in early-stage patients. AB - The propensity of myelodysplastic syndrome (MDS) to transform into acute myeloid leukemia (AML) suggests the existence of common pathogenic components for these malignancies. Here, four genes implicated in the development of AML were examined for promoter CpG island hypermethylation in cells from 37 patients with different stages of MDS. Aberrant methylation was detected by polymerase chain reaction amplification of bisulfite-treated DNA followed by denaturing gradient gel electrophoresis. The highest rate of methylation was found for p15INK4B (51%), followed by HIC1 (32%), CDH1 (27%), and ER (19%). Concurrent hypermethylation of > or = 3 genes was more frequent in advanced compared with early-stage MDS (P < or = 0.05), and hypermethylation of p15INK4B was associated with leukemic transformation in early MDS (P < or = 0.05). The median overall survival was 17 months for cases showing hypermethylation of > or = 1 genes vs. 67 months for cases without hypermethylation (P = 0.002). Specifically, promoter hypermethylation identified a subgroup of early MDS with a particularly poor prognosis (median overall survival 20 months vs. 102 months; P = 0.004). In multivariate analysis including stage and thrombocyte count, hypermethylation of > or = 1 genes was an independent negative prognostic factor (P < 0.05). These data suggest that hypermethylation of p15INK4B, HIC1, CDH1, and ER contribute to the development and outcome of MDS. PMID- 16343269 TI - Arg452 substitution of the erythroid-specific 5-aminolaevulinate synthase, a hot spot mutation in X-linked sideroblastic anaemia, does not itself affect enzyme activity. AB - Mutations of the erythroid-specific 5-aminolaevulinate synthase (ALAS2) gene are known to be responsible for X-linked sideroblastic anaemia (XLSA). An amino acid (AA) substitution for arginine at the 452 AA position of the ALAS2 protein is the most frequent mutation, which has been found in approximately one-quarter of patients with XLSA. Despite its high frequency, there has been no report on the enzymatic activity of Arg452 mutant proteins. In this study, we examined enzymatic activity in vitro of two Arg452 mutants, Arg452Cys and Arg452His, which were found in two new pedigrees of XLSA. While these mutations must be responsible for the clinical phenotype of XLSA in patients, the enzymatic activity and stability of these mutant proteins studied in vitro are indistinguishable from those of the wild type protein. These findings suggest that the Arg452 mutation of the ALAS2 gene by itself does not decrease the enzymatic activity or the stability in vitro, and that there may be an additional factor(s) in the bone marrow, which ensures the full ALAS2 activity in vivo. PMID- 16343270 TI - Histone deacetylase inhibitors reduce VEGF production and induce growth suppression and apoptosis in human mantle cell lymphoma. AB - OBJECTIVES: Mantle cell lymphoma (MCL) is an incurable disease with an aggressive course and novel treatment strategies are urgently needed. The purpose of this study was to evaluate the effects of histone deacetylase (HDAC) inhibitors, a new group of antiproliferative agents, on human MCL cells. METHODS: Three MCL cell lines (JeKo-1, Hbl-2 and Granta-519) were exposed to different concentrations of the HDAC inhibitors sodium butyrate (NaB) and suberoylanilide hydroxamic acid (SAHA) for 8-72 h. Their effects on cell viability, apoptosis induction and cell cycle proliferation were studied. Moreover, the influence of SAHA on the expression of cyclin D1, the cell cycle regulators p21 and p27 and the production of vascular endothelial growth factor (VEGF) were analyzed. RESULTS: The HDAC inhibitors induced accumulation of acetylated histones in MCL cells. MTT assays and Annexin-V staining showed that they potently inhibited viability in a dose dependent manner and induced apoptosis in all cell lines tested. Cell cycle analysis indicated that their exposure to SAHA or NaB decreased the proportion of cells in S phase and increased the proportion of cells in the G0/G1 and/or G2/M phases. Incubation with the two HDAC inhibitors resulted in downregulation of cyclin D1. SAHA lead to an upregulation of p21 in all cell lines and an upregulation of p27 in JeKo-1 and Granta-519 cells, while expression of p27 in Hbl-2 was not altered. In addition, SAHA inhibited the production of the angiogenic cytokine VEGF. Treatment with NaB increased the expression of p21 in JeKo-1 and Hbl-2 cells, while in Granta 519 cells no effect was noted. The expression of p27 remained constant in all three cell lines after exposure to NaB. CONCLUSION: Based on these findings, we provide evidence that HDAC inhibitors have antiproliferative effects in MCL and may represent a promising therapeutic approach. PMID- 16343271 TI - Serum insulin-like growth factor is not elevated in patients with early B-cell chronic lymphocytic leukemia but is still a prognostic factor for disease progression. AB - OBJECTIVES: Insulin-like growth factor 1 (IGF-1) is an important growth and anti apoptotic factor for the cancer cells in several malignancies and in multiple myeloma recent studies support the hypothesis of a role for IGF-1 in disease progression; however, clinico-biological relevance of IGF-1 was never studied in B-cell chronic lymphocytic leukemia (CLL). PATIENTS AND METHODS: Using a quantitative sandwich immunoassay technique (ELISA) (Quantikine, Human IGF-1 and IGFBP-3, R&D Systems), we measured the concentration of IGF-1 and its major binding protein IGF-binding protein 3 (IGFBP-3) in serum drawn at the time of diagnosis from 77 Binet stage A CLL patients. RESULTS: Either IGF-1 or IGFBP-3 were significantly decreased compared with healthy age- and sex-matched controls (P < 0.0001 for both; Mann-Whitney test). Serum levels of IGF-1 and IGFBP-3 paralleled each other (P = 0.002); in contrast, no significant correlation was found between serum levels of IGF-1 and clinico-hematological variables including age (P = 0.253), sex (P = 0.270), Rai clinical substages (P = 0.140), lactate dehydrogenase (P = 0.956), beta2-microglobulin (P = 0.368), lymphocyte count (P = 0.703) and lymphocyte doubling time (LDT, P = 0.233). When correlation were attempted with circulating levels of angiogenic cytokines such as vascular endothelial growth factor (P = 0.971), basic fibroblastic growth factor (P = 0.695), angiogenin (P = 0.282) or adhesion molecules such as vascular cell adhesion molecule-1 (P = 0.318), intercellular adhesion molecule-1 (P = 0.883) and platelet endothelial cell adhesion molecule-1 (P = 0.772) similar results were found. Serum levels of IGF-1 were further evaluated as a dichotomous variable with respect to progression-free survival (PFS), an endpoint surrogate for overall survival in early B-cell CLL. The best separation of curves was seen with the cutoff point at the 75th percentile of IGF-1 levels (i.e., 93 pg/mL). Median PFS was 63 months in the patient group with low IGF-1, compared with a median PFS of 40 months in the remaining patients (P = 0.03). In the multivariate analysis performed including variables significant at univariate analysis [i.e. Rai substage (P = 0.002); LDT (P = 0.004), IGF-1 (P = 0.01)], only Rai substage retained prognostic significance (P = 0.006). However, after removing from analysis LDT (only six of 77 had an LDT < 12 months), either IGF-1 or Rai substage entered the model at a significant level (P = 0.03 and P = 0.01, respectively). CONCLUSIONS: IGF-1 did not correlate with markers of tumor burden or clinical status in CLL thus suggesting that levels of this cytokine do not reflect the intrinsic malignancy of disease. Results of the present study highlight, however, its involvement in mechanisms of disease progression in early CLL. PMID- 16343272 TI - Follicular lymphoma in early stages: high risk of relapse and usefulness of the Follicular Lymphoma International Prognostic Index to predict the outcome of patients. AB - BACKGROUND AND OBJECTIVES: Patients with follicular lymphoma (FL) in advanced stages are currently deemed incurable with standard treatments. However, FL is considered to be eradicable in the small group of patients presenting with localized disease. The objective of this study was to analyze the clinical features and the outcome of a series of patients with FL in early stages with a long follow-up. PATIENTS AND METHODS: A total of 48 patients (25m/23f; median age: 50 yr) diagnosed consecutively with FL in Ann Arbor stage I (25 cases) or II (23) at a single institution with a median follow-up of 9.5 yr were included in the study. Main biological and clinical characteristics at diagnosis, including Follicular Lymphoma International Prognostic Index (FLIPI) were analyzed; treatment and response were assessed and analyzed for prognosis. RESULTS: The histologic subtypes were: FL type I, 20 cases (42%); type II, 24 (50%); type III, three (6%); and unclassifiable, one (2%). Distribution according to FLIPI was: low risk (36 cases) and intermediate risk (five cases). Treatment mainly consisted of combination chemotherapy (CHOP in 34 cases) plus involved-field radiotherapy in 26 cases. Forty patients (89%) achieved a complete response (CR), three (7%) a partial response, and two (4%) were non-responders; the remaining three patients did not receive therapy. No initial variable predicted CR achievement. About 57% of the patients in CR eventually relapsed with a relapse risk of 46% at 10 yr. Intermediate-risk FLIPI predicted failure-free survival. Histologic transformation was observed in six patients with a 10-yr risk of transformation of 13%. Twelve patients died during follow-up, in two cases as a result of unrelated causes. Overall survival (OS) at 10 yr was 79%. The FLIPI was the sole variable predicting OS. CONCLUSIONS: Although the majority of patients with localized FL achieve CR, the risk of relapse is high. The FLIPI is of prognostic value in these patients. PMID- 16343273 TI - Potential mechanism of resistance to TRAIL-induced apoptosis in Burkitt's lymphoma. AB - OBJECTIVES: Members of the tumor necrosis factor family are potent inducers of apoptosis in sensitive cells and may be suitable for novel anti-cancer therapies aimed at inducing apoptosis via the activation of receptors with the death domain on malignant cells. We characterized the sensitivity of Burkitt's lymphoma (BL) cell lines to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and anti-Fas agonist, and investigated the mechanism of resistance of BL cell lines to TRAIL and Fas apoptotic pathways. METHODS: Epstein-Barr virus (EBV) status in BL cell lines was determined by PCR. The extent of apoptosis following exposure to TRAIL and anti-Fas agonist was measured by 7A6 antigen staining. Expression of TRAIL receptors and Fas was determined by flow cytometry and reverse transcriptase-PCR. Western blot analyses were used to determine the expression of proapoptotic and antiapoptotic proteins. NF-kappaB activity was evaluated by electrophoretic mobility shift assay. RESULTS: The sensitivity of BL cell lines to anti-Fas agonist depended on the expression of Fas. In contrast, the expression of TRAIL receptors did not correlate with the sensitivity to TRAIL induced apoptosis. Interestingly, EBV-infected BL cell lines which showed constitutive levels of NF-kappaB activation, were TRAIL-resistant. NF-kappaB inhibitors reversed the resistance to TRAIL-induced apoptosis. CONCLUSIONS: Our results suggest that activation of NF-kappaB by EBV infection plays an important role in resistance of BL cell lines to TRAIL-induced apoptosis, and that NF kappaB inhibitors may be useful adjuncts in clinical use of TRAIL against BL. PMID- 16343274 TI - Steroid-responsive functional B12 deficiency in association with transcobalamin II polymorphism 776C --> G. AB - We present a case of intracellular vitamin B12 deficiency presenting with confusion, subacute combined degeneration of the cord, megaloblastic anaemia and intrinsic factor antibodies in the serum. Diagnosis was delayed by a normal serum B12 level and was confirmed by a grossly elevated serum homocysteine. There was a dramatic response to steroids. The patient was heterozygous for the transcobalamin (TC) II polymorphism 776C --> G. This case demonstrates the importance of functional assessment of intracellular B12 activity (e.g. serum homocysteine) in excluding B12 deficiency, the role of steroids in pernicious anaemia and a possible clinical correlation of a TCII polymorphism. PMID- 16343275 TI - Hb Woodville, a rare alpha-globin variant, caused by codon 6 mutation of the alpha1 gene. AB - Since 1995, the national programme for the prevention and control of severe thalassaemia has been implemented in Thailand. This programme is composed of the population screening in pregnant women and couples by osmotic fragility, HbE screening and the confirmation test using haemoglobin analyses by electrophoresis or chromatography. Thereafter, several hitherto unidentified haemoglobins (Hbs) with structural defects are increasingly described and these variants are now easily studied using DNA technology. In this study, the authors describe the haematology and molecular analyses in a 28-yr-old healthy female who was identified as having an exceptionally 'high HbA2' from haemoglobin analysis. Subsequent analyses demonstrated that observed atypical 'HbA2' was, in fact, a rare innocuous alpha-globin variant, called Hb Woodville [alpha 2 6(A4); Asp --> Tyr]. For the first time, this abnormal Hb species is characterised at the molecular level. PMID- 16343276 TI - Development of rapidly progressive liver light chain deposition under VAD chemotherapy in multiple myeloma. AB - Light chain deposition disease (LCDD) is a multisystemic disorder seen in the setting of plasma cell dyscrasias. The histological characteristic of this disorder is the deposition of a homogeneous, granular, slightly eosinophilic and non-Congophilic material that shows immunostaining for monoclonal light chains (kappa or gamma), while in primary amyloidosis (AL) the proteinaceous substance is fibrillar and Congo red positive. In contrast with AL, the light chain in LCDD is usually of the kappa-type. Renal involvement, resulting in nephrotic syndrome, is usually the prominent feature of LCDD. Patients with this disease may also have heart, liver or other organ involvement, mimicking the picture of primary systemic amyloidosis. However, liver failure has rarely been described in patients with LCDD. A patient with myeloma-associated LCDD who developed rapidly progressive liver kappa light chain deposition with fatal outcome after undergoing the first cycle of vincristine/doxorubicin/dexamethasone chemotherapy is reported. PMID- 16343277 TI - Concomitant hairy cell and acute myeloid leukemia. AB - Secondary malignancies in patients with hairy cell leukemia (HCL) are well described and treatment of HCL is discussed as their cause. We describe a 58-yr old patient who presented with both acute myeloid leukemia (AML) and HCL at the same time. Treatment with cladribine, daunorubicin and cytosine arabinoside, followed by autologous stem cell transplantation, induced complete remission of AML and hematologic remission of HCL for 22 months, when he relapsed with AML. This concomitant occurrence of AML and HCL is suggestive of a genetic predisposition rather then coincidence or relation to purine analoga. PMID- 16343278 TI - Acquired thrombasthenia due to GPIIbIIIa platelet autoantibodies in a 4-yr-old child. PMID- 16343281 TI - Pharmacokinetics of cefepime administered by i.v. and i.m. routes to ewes. AB - The pharmacokinetics of cefepime were studied following i.v. and i.m. administration of 20 mg/kg in 10 ewes. Following i.v. administration of a single dose, the plasma concentration-time curves of cefepime were best fitted using a two-compartment open model. The elimination half-life (t(1/2beta)) was 1.76 +/- 0.07 h, volume of distribution at steady-state [V(d(ss))] was 0.32 +/- 0.01 L/kg and total body clearance (Cl(B)) was 2.37 +/- 0.05 mL/min.kg. Following i.m. administration, the drug was rapidly absorbed with an absorption half-life (t(1/2ab)) of 0.49 +/- 0.05 h, maximum plasma concentration (Cmax) of 31.9 +/- 1.5 mug/mL was attained at (tmax) 1.1 +/- 0.2 h and the drug was eliminated with an elimination half-life (t(1/2el)) of 2.06 +/- 0.11 h. The systemic bioavailability (F) after i.m. administration of cefepime was 86.8 +/- 7.5%. The extent of plasma protein binding measured in vitro was 14.8 +/- 0.54%. The drug was detected in urine for 36 h postadministration by both routes. PMID- 16343282 TI - Investigation of Japanese quail (Coturnix japonica) as a pharmacokinetic model for cockatiels (Nymphicus hollandicus) and Poicephalus parrots via comparison of the pharmacokinetics of a single intravenous injection of oxytetracycline hydrochloride. AB - The purpose of this study was to determine whether Japanese quail (Coturnix japonica) would serve as a pharmacokinetic animal model for two small companion parrots: cockatiels (Nymphicus hollandicus) and Poicephalus parrots. Oxytetracycline (OTC) was the pharmacologic agent chosen for this study as it is eliminated primarily by renal glomerular filtration and undergoes minimal metabolism. A single intravenous injection of 20 mg/kg oxytetracycline hydrochloride was administered to the three study groups and blood samples were obtained at 5, 10, 15, and 30 min post-OTC injection as well as 1, 2, 4, 8, 12 and 24 h post-OTC injection. Quantification of plasma OTC was accomplished using a standardized microbial inhibition assay. Naive-pooled data (NPD) analysis of the plasma concentration-time profile of OTC best fit a two-compartment open model for all three avian species. Noncompartmental analysis of the mean data yielded the following parameters for quail, cockatiels and Poicephalus parrots respectively: lambda(z) = 3.14, 4.57, 3.71 h; AUC = 38.9, 42.7, 49.6 microg x h/mL; and Cl = 514, 468, 403 mL/h/kg. Based on the similarity of these pharmacokinetic parameters, it appears that quail could be used as a model species to predict the appropriate OTC dosing regimen for small psittacine birds. A bootstrap procedure was also applied to these sparse data sets for both compartmental and noncompartmental analysis. The bootstrap procedure allowed for the calculation of variability of parameters; however, the estimates of the parameters were very similar to those calculated using the NPD and the data mean values. PMID- 16343283 TI - An original system for the continuous monitoring of antimicrobial use in poultry production in France. AB - Data collection at the flock level on a continuous basis, for antimicrobial use surveillance purposes, provides a complex challenge. Using existing on-farm records, produced as part of the regulations for meat inspection, a sampling scheme was implemented to collect on a simple, representative and continuous basis, the forms providing history of antimicrobial consumption of poultry flocks slaughtered in Brittany. Relational database-stored information provided by the forms includes treatment purpose, antimicrobial drug administered, age at treatment and treatment duration. The aim of this study was to collect information describing antimicrobial drug use quantitatively and qualitatively, by computing the frequency of antimicrobial use according to species, therapeutic class, treatment purpose, duration, and animal age at treatment. A study was carried out to check the validity of farmers' declarations. Examples of data and results are presented such as the relative importance of the different antimicrobial classes in relation to the live weight treated, which shows that beta-lactams and tetracyclines are the main classes used in broilers and turkeys respectively. Average bird weight at treatment was 0.5 kg for a broiler and 3 kg for a turkey. Nevertheless, several 'at-risk' periods could be identified. Initial results after 1 year of data collection showed that this surveillance system provides valid, detailed and useful data about antimicrobial drug use in poultry production at a low cost and with only a moderate time involvement. PMID- 16343284 TI - PK and PK/PD of doxycycline in drinking water after therapeutic use in pigs. AB - A commercial doxycycline formulation was administered in drinking water to 12 pigs at the recommended dose of 10 mg/kg daily for 5 days. The mean plasma concentration at steady-state was 1.37 +/- 1.21 microg/mL, which was reached at 68 +/- 27.2 h postadministration. Absorption and elimination half-life values were 7.20 +/- 2.42 and 7.01 +/- 2.10 h, respectively. Most plasma concentrations during dosing were higher than the minimum inhibitory concentrations (MICs) described for the main porcine bacterial pathogens of the respiratory tract (Pasteurella multocida, Actinobacillus pleuropneumoniae, Bordetella bronchiseptica and Mycoplasma hyopneumoniae). It is concluded that when pigs were treated with doxycycline in drinking water at the recommended rate, therapeutically effective concentrations were achieved throughout the treatment period, supporting the clinical use of this tetracycline in the control of respiratory infections. However, inter-animal differences were marked. PMID- 16343285 TI - Inhibitory effects of the beta-adrenergic receptor agonist zilpaterol on the LPS induced production of TNF-alpha in vitro and in vivo. AB - In this study the anti-inflammatory properties of zilpaterol, a beta2-adrenergic receptor (AR) agonist specifically developed as a growth promoter in cattle were investigated. Although zilpaterol has a different structure compared with the beta2-AR agonists known to date, it was noted that it was able to bind to both the beta2-AR (Ki = 1.1 x 10(-6)) and the beta1-AR (Ki = 1.0 x 10(-5)). Using lipopolysaccharide (LPS)-exposed U937 macrophages, the production of cyclic adenosine-3',5'-cyclic monophosphate (cAMP) and tumor necrosis factor alpha (TNF alpha) were investigated. Zilpaterol inhibited TNF-alpha release and induced intracellular cAMP levels in a dose-dependent manner. The inhibition of TNF-alpha release and induction of cAMP production was mainly mediated via the beta2-AR, as indicated by addition of beta1- and beta2-specific antagonists. The effects of zilpaterol were investigated in LPS-treated male Wistar rats after pretreatment with zilpaterol. Zilpaterol dosed at 500 microg/kg body weight reduced the TNF alpha plasma levels. In conclusion, zilpaterol is a beta2-adrenergic agonist and an inhibitor of TNF-alpha production induced by LPS both in vivo and in vitro. PMID- 16343286 TI - Pharmacokinetics of oral omeprazole in llamas. AB - Gastrogard, an oral formulation of omeprazole, was given to six llamas at a dose of 4 mg/kg once a day for 6 days. Plasma samples were collected at 0, 15, 30, 45, and 60 min and 2, 3, 4, 6, 8, 12, and 24 h on days 1 and 6. Plasma omeprazole concentrations were measured by high-pressure liquid chromatography with ultraviolet detection. Pharmacokinetic parameters calculated included the area under the curve (AUC(0-infinity)), peak plasma concentration (Cmax), time of peak plasma concentration (Tmax), and terminal half-life (t(1/2)). On day 6, plasma omeprazole concentrations reached a Cmax of 0.12 microg/mL at a Tmax of 45 min. The t(1/2) of omeprazole was 2.3 h and the AUC(0-infinity) was 0.38 h x microg/mL. Plasma concentrations remained above the minimum concentration for inhibition of gastric acid secretion projected from other studies on day 6 in all the llamas for approximately 6 h. However, the AUC(0-infinity) was below the concentrations associated with clinical efficacy. It was not possible to measure oral systemic bioavailability because there was no i.v. data collected from these animals. However, using data published on the i.v. pharmacokinetics of omeprazole in llamas, oral absorption was estimated to be only 2.95%. Due to low absorption the oral dose was increased to 8 and 12 mg/kg and studies were repeated. There were no significant differences in Cmax, Tmax, or AUC(0-infinity) for either of the increased doses. These results indicate that after 6 days of treatment with doses up to 12 mg/kg, oral omeprazole produced plasma drug concentrations which are not likely to be associated with clinical efficacy in camelids. PMID- 16343287 TI - Frequency of the nt230 (del4) MDR1 mutation in Collies and related dog breeds in Germany. AB - MDR1 (ABCB1) P-glycoprotein exerts a protective function in the blood-brain barrier thereby limiting the entry of many drugs and other xenobiotics to the central nervous system. A nonsense mutation has been described for Collies and related dog breeds which abolishes this function and is associated with increased susceptibility to neurotoxic side effects of several drugs including ivermectin, moxidectin and loperamide. In order to evaluate the occurrence and frequency of this nt230 (del4) MDR1 mutation in Germany, we screened 1500 dogs. Frequency of the homozygous mutated genotype was highest for Collies (33.0%), followed by Australian Shepherd (6.9%) and Shetland Sheepdog (5.7%). Thirty-seven percent of the Waller dogs and 12.5% of the Old English Sheepdogs were heterozygous for the mutant MDR1 (-) allele. Considering the predominant role of MDR1 P-glycoprotein in drug disposition and in particular for blood-brain barrier protection, MDR1 genotype-based breeding programs are recommended for improving the safety of drug therapy in these canine breeds. PMID- 16343288 TI - Inhibitory effect of several fluoroquinolones on hepatic microsomal cytochrome P 450 1A activities in dogs. AB - We examined inhibitory effects of ofloxacin (OFX), orbifloxacin (OBFX), ciprofloxacin (CFX), enrofloxacin (EFX) and norfloxacin (NFX) on cytochrome P-450 1A (CYP1A) activities using hepatic microsomes from four beagle dogs. Ethoxyresorufin O-de-ethylation was referred as CYP1A activities. All the fluoroquinolones inhibited the reaction in a noncompetitive manner. The determined inhibitory constants were the followings; 10.1 +/- 3.8 mM for OFX, 6.43 +/- 2.01 mM for OBFX, 0.726 +/- 0.134 mM for CFX, 4.06 +/- 1.19 mM for EFX and 4.75 +/- 1.63 mM for NFX respectively. As these values are >100-fold of plasma concentrations after a clinical single dose of the fluoroquinolones, it is suggested that the inhibitory effect on CYP1A activities is not so high to elicit drug-drug interaction with CYP1A substrates, when these fluoroquinolones are co administered. Mechanism based inhibition was also examined in this study. Of the five fluoroquinolones examined, OFX, OBFX and CFX had this inhibition manner. As this inhibition is irreversible, inhibitory effects of the three fluoroquinolones may accumulate, when they are repeatedly administered. Therefore, OFX, OBFX and CFX may result in substantial drug-drug interaction with a CYP1A substrate even in clinical states. As EFX is metabolized to CFX in the body, it may also have the same possibility. PMID- 16343289 TI - Quantitative analysis of the effect of probenecid on pharmacokinetics of 99mTc mercaptoacetyltriglycine in dogs. AB - Effect of probenecid on pharmacokinetics of 99mTc-mercaptoacetylytriglycine (99mTc-MAG3) in dogs was investigated before (control), and after 15 min and 24 h of i.v. injection of probenecid (20 mg/kg). Plasma concentration-time profiles of 99mTc-MAG3 were described with a two-compartment open model. Plasma 99mTc-MAG3 clearances (Clp, ml/min/kg) were 7.9 +/- 0.5, 3.3 +/- 0.5 and 4.8 +/- 1.3 in control, 15 min and 24 h after probenecid administration respectively. Similarly, the biological half-lives at elimination phase (t(1/2), h) were 0.61 +/- 0.09, 0.79 +/- 0.11 and 0.74 +/- 0.12, and volumes of distribution at steady state (Vdss, L/kg) were 0.29 +/- 0.04, 0.20 +/- 0.05 and 0.25 +/- 0.06 respectively. The prolonged biological half-life and decreased Vdss decreased Clp significantly. Clp was a function of plasma probenecid concentration based on Michaelis-Menten kinetics. The maximum Clp inhibition (Imax) by probenecid and the plasma probenecid concentration that induced 50% of Imax (I50) were estimated to be 72 +/- 12% and 13 +/- 8 microg/ml respectively. This means that the rest (about 28%) of the Clp is not blocked by probenecid alone, suggesting the possibility of another route(s) of elimination or renal transporters which are independent from probenecid. Moreover, inter-species correlation between Clp of 99mTc-MAG3 and body weight are discussed. PMID- 16343290 TI - In vitro effects of bethanechol on equine gastrointestinal contractility and functional characterization of involved muscarinic receptor subtypes. AB - The goal of this study was to investigate the effect of bethanechol (BeCh) on contractility patterns of smooth muscle preparations of equine duodenum descendens, jejunum, caecum and pelvic flexure in vitro. Concentration-response relationships were developed for BeCh using in vitro assays with and without preincubation of muscarinic (M) receptor antagonists for M2 and M3 receptors. BeCh induced a significant, concentration-dependent increase in contractile response in equine intestine in specimens with circular orientation. The maximal effect was largest for jejunal specimens with no difference in EC50 within the different locations investigated. The M2 antagonist, AF-DX 116, caused a rightward shift of the concentration-response curve and the M3 antagonist, 4-DAMP (1,1-dimethyl-4-diphenylacetoxypiperidinium iodide), almost completely inhibited the effect of BeCh over the entire concentration-response curve. These data provide evidence that, although the effect of BeCh is predominantly mediated by M3 receptors, M2 muscarinic receptors also play a role in BeCh-induced contraction in specimens of equine intestine. The involvement of other muscarinic receptor subtypes cannot be excluded. Further studies are necessary to understand the effect of BeCh in vivo including diseased animals. PMID- 16343291 TI - Pharmacokinetics and oral bioavailability of pentoxyfylline in broiler chickens. AB - The pharmacokinetic properties of pentoxyfylline and its metabolites were determined in healthy chickens after single intravenous and oral dosage of 100 mg/kg pentoxyfylline. Plasma concentrations of pentoxyfylline and its metabolites were determined by a validated high-performance liquid chromatographic method. After intravenous (i.v.) and oral (p.o.) administration, the plasma concentration time curves were best described by a one-compartment open model. The mean elimination half-life (t(1/2el)) of pentoxyfylline was 1.05 h, total body clearance 1.90 L/h x kg, volume of distribution 2.40 L/kg and the mean residence time was 2.73 h, after i.v. administration. After oral dosing, mean maximal plasma concentration of pentoxyfylline was 4.01 microg/mL and the interval from p.o. administration until maximum concentration was 1.15 h. The mean oral bioavailability was found to be 28.2%. Metabolites I, IV and V were present in chicken plasma after both i.v. and p.o. administration, with metabolite V being the most dominant. PMID- 16343292 TI - Effect of tepoxalin on renal function in healthy dogs receiving an angiotensin converting enzyme inhibitor. AB - The objective of this study was to investigate renal function in clinically normal dogs receiving tepoxalin, a nonsteroidal inflammatory drug, either in association with or without an angiotensin-converting enzyme inhibitor (ACEI). Ten adult female Beagle dogs were used in the three phases of the study. The dogs were administered the drugs once daily for 7 days (experiment 1: placebo/tepoxalin/tepoxalin and benazepril; experiment 2: enalapril/tepoxalin and enalapril) or for 28 days (experiment 3: tepoxalin and benazepril together). Renal function was assessed by measurement of glomerular filtration rate (GFR) by renal scintigraphy [(renal uptake of 99mTc-diethylenetriaminepentacetic acid (DTPA)] and plasma clearance of 99mTc-DTPA. Compared with the placebo group, renal uptake and plasma clearance of 99mTc-DTPA were not significantly modified after a 7-day period of treatment with tepoxalin or enalapril alone, tepoxalin and benazepril or tepoxalin and enalapril together. No significant change was obtained in GFR after a 28-day period of dosing with tepoxalin and benazepril together. Therefore, it was concluded that tepoxalin did not alter renal function in healthy Beagle dogs receiving ACEI. PMID- 16343293 TI - Pharmacodynamic study of a long-acting parenteral formulation of omeprazole in horses. PMID- 16343294 TI - Dementia: new projections and time for an updated response. PMID- 16343295 TI - Projections of future numbers of dementia cases in Australia with and without prevention. AB - OBJECTIVE: To produce projections of the number of dementia cases in Australia from 2000 to 2050. These projections examine the effects of an ageing population on number of dementia cases and also the potential impact of preventive strategies that delay age of dementia onset. METHOD: Data from several meta analyses of dementia prevalence and incidence were combined with age-specific population projections to arrive at estimates of future numbers of cases. A statistical model of dementia incidence was developed and used to estimate the effects of delaying onset by up to 5 years. RESULTS: Without prevention, prevalence of dementia is estimated to increase from 172 000 in 2000 to 588 000 in 2050. Over the same period, the incidence of dementia is estimated to increase from 43 000 to 143 000 new cases a year. Delaying onset by 5 years would decrease prevalence in 2050 by 44%. Even a 6-month delay would reduce prevalence by 6%. CONCLUSIONS: Ageing of the population will lead to a rapid increase in number of dementia cases and an increasing burden for the working-age population. However, even modest prevention efforts could lessen the impact. PMID- 16343296 TI - Mapping the onset of psychosis: the Comprehensive Assessment of At-Risk Mental States. AB - OBJECTIVE: Recognizing the prodrome of a first psychotic episode prospectively creates the opportunity of intervention, which could delay, ameliorate or even prevent onset. Valid criteria and a reliable methodology for identifying possible prodromes are needed. This paper describes an instrument, the Comprehensive Assessment of At-Risk Mental States (CAARMS), which has been designed for such a purpose. It has two functions: (i) to assess psychopathology thought to indicate imminent development of a first-episode psychotic disorder; and (ii) to determine if an individual meets criteria for being at ultra high risk (UHR) for onset of first psychotic disorder. This paper describes the pilot evaluation of the CAARMS. METHOD: Several methodologies were used to test the CAARMS. First, CAARMS scores in a group of UHR young people and the association between CAARMS scores and the risk of transition to psychotic disorder, were analysed. Second, CAARMS scores in a UHR group were compared to a control group. To assess concurrent validity, CAARMS-defined UHR criteria were compared to the existing criteria for identifying the UHR cohort. To assess predictive validity, the CAARMS-defined UHR criteria were applied to a sample of 150 non-psychotic help-seekers and rates of onset of psychotic disorder at 6-month follow-up determined for the CAARMS positive (i.e. met UHR criteria) group and the CAARMS-negative (i.e. did not meet UHR criteria) group. The inter-rater reliability of the CAARMS was assessed by using pairs of raters. RESULTS: High CAARMS score in the UHR group was significantly associated with onset of psychotic disorder. The control group had significantly lower CAARMS scores than the UHR group. The UHR criteria assessed by the CAARMS identified a similar group to the criteria measured by existing methodology. In the sample of non-psychotic help-seekers those who were CAARMS positive were at significantly increased risk of onset of psychotic disorder compared to those who were CAARMS-negative (relative risk of 12.44 (95% CI = 1.5 103.41, p = 0.0025)). The CAARMS had good to excellent reliability. CONCLUSIONS: In these preliminary investigations, the CAARMS displayed good to excellent concurrent, discriminant and predictive validity and excellent inter-rater reliability. The CAARMS instrument provides a useful platform for monitoring subthreshold psychotic symptoms for worsening into full-threshold psychotic disorder. PMID- 16343297 TI - Integrating approaches to psychotherapy in psychosis*. AB - OBJECTIVE: The evidence base for specific psychological treatments for psychosis is now well established, but many practitioners see themselves as integrationist in approach. The basic tenets of integration are explored with an emphasis on understanding how different levels of need can be conceptualized and then used to 'adapt' a treatment to meet those needs in an individual. The needs are then incorporated into an integrated treatment formulation. METHOD: The evidence base is strongest for cognitive behavioural and family approaches, but the present paper summarizes concepts from two specific models of therapy that are intrinsically integrational in their approach: cognitive analytic therapy and psychodynamic interpersonal therapy. RESULTS: Both approaches show aspects of integration. However, following this approach to integration to its limit would ultimately lead to one undifferentiated therapy. CONCLUSIONS: Both approaches share a common set of values of developing specific ways of increasing collaboration and working together, and these values are shown to underpin adaptive ways of working with psychosis, but further critical analysis of the development of integrative models is needed. PMID- 16343298 TI - Relationships between self-reported potentially traumatizing events, psychoform and somatoform dissociation, and absorption, in two non-clinical populations. AB - OBJECTIVE: Some authors have suggested that the personality characteristic 'fantasy proneness' may mediate the correlation between reported potentially traumatizing events and dissociative symptoms. Other authors question the reported magnitude of this correlation in non-clinical samples, because these are usually derived from student samples and may therefore suffer from a restriction of range. The primary aim of this study is to assess the relationship between a self-report measure of traumatization and psychoform dissociation as well as somatoform dissociation in a non-clinical population, while accounting for the influence of fantasy proneness. METHOD: Two random non-clinical samples, that is, a student and an adult non-student sample, completed a range of relevant self report questionnaires. Absorption was used as an index of fantasy proneness. RESULTS: The range of reported potentially traumatizing events was restricted in students, compared to non-students. In both samples a significant correlation was found between reported potentially traumatizing events and dissociation. After partialling out absorption, the relationship between reported potential traumatization and psychoform dissociation diminished substantially in both samples. The magnitude of the correlation with somatoform dissociation decreased to a lesser degree, so that it remained significant in both samples. CONCLUSIONS: The correlation between somatoform dissociation and reported traumatization, after partialling out absorption, gives a reliable estimate of the magnitude of the relationships between potentially traumatizing events and dissociation. Findings regarding traumatization and dissociation in students should be generalized to the general population cautiously. PMID- 16343299 TI - Australia's National Mental Health Strategy and deinstitutionalization: some empirical results. AB - OBJECTIVE: To determine the role of the National Mental Health Strategy in the deinstitutionalization of patients in psychiatric hospitals in Queensland. METHOD: Regression analysis (using the maximum likelihood method) has been applied to relevant time-series datasets on public psychiatric institutions in Queensland. In particular, data on both patients and admissions per 10 000 population are analysed in detail from 1953-54 to the present, although data are presented from 1883-84. RESULTS: These Queensland data indicate that deinstitutionalization was a continuing process from the 1950s to the present. However, it is clear that the experience varied from period to period. For example, the fastest change (in both patients and admissions) took place in the period 1953-54 to 1973-74, followed by the period 1974-75 to 1984-85. CONCLUSIONS: In large part, the two policies associated with deinstitutionalization, namely a discharge policy ('opening the back door') and an admission policy ('closing the front door') had been implemented before the advent of the National Mental Health Strategy in January 1993. Deinstitutionalization was most rapid in the 30-year period to the early 1980s: the process continued in the 1990s, but at a much slower rate. Deinstitutionalization was, in large part, over before the Strategy was developed and implemented. PMID- 16343300 TI - Identifying marginal housing for people with a mental illness living in rural and regional areas. AB - OBJECTIVE: Homelessness among people with a mental illness is a serious issue. Objective: The study aimed to identify the types of housing available for people with a mental illness in the region; to develop an audit tool to assess the housing of patients of the mental health service; and to pilot this audit tool. METHOD: Key informants were interviewed to obtain information about the type of housing options available in the Loddon Campaspe Southern Mallee Region. This information was used to develop a survey to audit the housing status of existing patients of the area mental health service. A pilot study using the survey was completed by case managers for 81 patients who were being case managed by the area mental health service. RESULTS: There were a wide variety of housing options in the region, but housing availability was not evenly distributed. Although most patients lived in owned or rented accommodation, a substantial proportion of patients had difficulties with accessing the required services, the affordability of their housing, uncertainty of tenure or were at risk of violence within their housing. CONCLUSIONS: A substantial proportion of patients treated by a rural area mental health service had lived in impoverished housing. PMID- 16343301 TI - Hurly-burly of psychiatric ethics. AB - This is the introductory paper to the special issue on ethics in psychiatry. We introduce the other papers that follow and set them in a context. Inevitably, they represent only a thin slice of the work going on in psychiatric ethics. But they serve to show two unique features of this discipline. First, it has a tendency to dig deep and to make connections with other philosophical concepts. So, for example, in a number of ways the papers that follow touch on the nature of personhood. We examine this notion. Second, psychiatric ethics, because of its content and its embededness in the real world, tends to hit upon diverse and sometimes conflicting values. We introduce the idea of values-based medicine, which provides both a theoretical framework and a practical approach to the common dilemmas of psychiatric practice. The need to think deeply, but also clearly and coherently, combined with the need to engage with the hurly-burly of the world of patients, users and carers, suggests the reasons why psychiatric ethics offers a paradigm for practical ethics generally. PMID- 16343302 TI - Mental health practice: can philosophy help? AB - Traditionally, philosophy has been regarded as operating at a level above practical application. However, the discipline can offer mental health policy and practice some valuable aid. First, its logical rigour can help to clarify concepts and expose inconsistency and prejudice; second, its wealth of theory about morality can enrich the concepts that guide mental health practice. By avoiding simplistic solutions (such as reliance on the 'four principles' of biomedical ethics) those who turn to the discipline of philosophy may learn how to be more critical of accepted policies and practices. PMID- 16343303 TI - Justice and welfare: two ethical paradigms in forensic psychiatry. AB - Forensic psychiatry, as a medical specialty, perhaps understandably leans toward beneficence or welfare as its main ethical underpinning. However, the special nature of the art or science of forensic psychiatry makes it imperative that beneficence is not the only ethical principle that guides the 'good' forensic psychiatrist. Indeed, the commonest ethical dilemmas in forensic psychiatry arise from a conflict between two ethical principles: beneficence, or promotion of welfare, and respect for justice. These two paradigms dominate discussions about the moral role and ethical duties of forensic psychiatrists and, in effect, give rise to two different practices in forensic psychiatry, each of which can also be said to have acquired a national identity. We discuss these competing principles and offer some thoughts about what this means for the ethics and values of forensic psychiatry. PMID- 16343304 TI - Moral responsibility, consciousness and psychiatry. AB - This article discusses the connection between consciousness and responsibility. Moral responsibility plays a crucial, but often implicit role in psychiatry in that it is often a therapeutic aim as well as an important evaluative concept. This article explains one of the more influential 'psychological' theories of moral responsibility, developed by Harry Frankfurt. In the second part of this article, a modified version of this account is applied to a range of psychiatric phenomena. PMID- 16343305 TI - Narrative, ethics and people with severe mental illness. AB - Starting from the premise that people are essentially narrative beings, I argue that the onset of severe mental illness compromises the narrative enterprise of being able to construct one's Self and one's relationships in meaningful and coherent ways. This is due to both the curtailment of opportunities for narrative engagement and the dispossession of those whose narratives do not conform to the current conceptualization of narrative and narrativity. In these circumstances, supporting the narrative enterprise is an ethical endeavour that requires that we examine not only which narratives we construct, but also how we construct them. This requires a re-thinking of what might constitute narrative and how we might facilitate or enhance the narrativity of people with severe mental illness. Following this, I suggest four means to support the narrativity of people with severe mental illness: through maintaining narrative continuity, maintaining narrative agency, countering master narratives and attention to small stories. PMID- 16343306 TI - Capacity for decision-making in Alzheimer's disease: selfhood, positioning and semiotic people. AB - The aim of this article is to explore some of the ethical issues surrounding the assessment and determination of capacity of people with dementia in light of their meaning-making ability as shown through discourse. Discourse analysis, two illustrative cases, social construction theory and positioning are used to highlight some of the ethical dilemmas involved in basing a determination of capacity on the diagnosis of dementia and neuropsychological tests of cognitive function. Although neuropsychological tests have their place in assessing some aspects of cognitive function, aspects cognition such as meaning-making ability and selfhood cannot be assessed in a standard format. In dementia, there can be a differential impairment of recall memory while the personality, values and substantial long-term memory remain intact, as does implicit memory for recent events. People with dementia are vulnerable to being negatively positioned, thereby unfairly undermining their rights to make decisions about aspects of their lives. Assessing the capacity of a person with dementia to engage in decision-making is presently in need of examination so as to take into account the person's meaning-making ability and selfhood. Incorrect negative positioning, based on the diagnosis and defects in recall memory, can obscure intact cognitive abilities that allow a person to make decisions about aspects of living, creating the possibility of lasting harm being inflicted on the person with dementia. PMID- 16343307 TI - Gregory Bateson (1904-1980): a reappraisal. PMID- 16343308 TI - Acute dystonia with intramuscular olanzapine in adolescent mania: a report. PMID- 16343309 TI - Value of the single case report. Marijuana, vomiting and compulsive bathing: a new clinical syndrome? PMID- 16343310 TI - Efficacy of transcranial magnetic stimulation in panic disorder: a case report. PMID- 16343311 TI - Re: mortality and mental illness. PMID- 16343316 TI - Interactions between arbuscular mycorrhizal fungi and bacteria and their potential for stimulating plant growth. AB - Arbuscular mycorrhizal (AM) fungi and bacteria can interact synergistically to stimulate plant growth through a range of mechanisms that include improved nutrient acquisition and inhibition of fungal plant pathogens. These interactions may be of crucial importance within sustainable, low-input agricultural cropping systems that rely on biological processes rather than agrochemicals to maintain soil fertility and plant health. Although there are many studies concerning interactions between AM fungi and bacteria, the underlying mechanisms behind these associations are in general not very well understood, and their functional properties still require further experimental confirmation. Future mycorrhizal research should therefore strive towards an improved understanding of the functional mechanisms behind such microbial interactions, so that optimized combinations of microorganisms can be applied as effective inoculants within sustainable crop production systems. In this context, the present article seeks to review and discuss the current knowledge concerning interactions between AM fungi and plant growth-promoting rhizobacteria, the physical interactions between AM fungi and bacteria, enhancement of phosphorus and nitrogen bioavailability through such interactions, and finally the associations between AM fungi and their bacterial endosymbionts. Overall, this review summarizes what is known to date within the present field, and attempts to identify promising lines of future research. PMID- 16343317 TI - Identification and characterization of ectosymbionts of distinct lineages in Bacteroidales attached to flagellated protists in the gut of termites and a wood feeding cockroach. AB - Bacterial attachments to nearly the entire surface of flagellated protists in the guts of termites and the wood-feeding cockroach Cryptocercus are often observed. Based on the polymerase chain reaction-amplified 16S rRNA gene sequences, we investigated the phylogenetic relationships of the rod-shaped, attached bacteria (ectosymbionts) of several protist species from five host taxa and confirmed their identity by fluorescence in situ hybridizations. These ectosymbionts are affiliated with the order Bacteroidales but formed three distinct lineages, each of which may represent novel bacterial genera. One lineage consisted of the closely related ectosymbionts of two species of the protist genus Devescovina (Cristamonadida). The second lineage comprised three phylotypes identified from the protist Streblomastix sp. (Oxymonadida). The third lineage included ectosymbionts of the three protist genera Hoplonympha, Barbulanympha and Urinympha in the family Hoplonymphidae (Trichonymphida). The ultrastructural observations indicated that these rod-shaped ectosymbionts share morphological similarities of their cell walls and their point of attachment with the protist but differ in shape. Elongated forms of the ectosymbionts appeared in all the three lineages. The protist cells Streblomastix sp. and Hoplonympha sp. display deep furrows and vane-like structures, but these impressive structures are probably evolutionarily convergent because both the host protists and their ectosymbionts are distantly related. PMID- 16343318 TI - Trophic regulation of Vibrio cholerae in coastal marine waters. AB - Cholera disease, caused by the bacterium Vibrio cholerae, afflicts hundreds of thousands worldwide each year. Endemic to aquatic environments, V. cholerae's proliferation and dynamics in marine systems are not well understood. Here, we show that under a variety of coastal seawater conditions V. cholerae remained primarily in a free-living state as opposed to attaching to particles. Growth rates of free-living V. cholerae (micro: 0.6-2.9 day(-1)) were high (similar to reported values for the bacterial assemblages; 0.3-2.5 day(-1)) particularly in phytoplankton bloom waters. However, these populations were subject to heavy grazing-mortality by protozoan predators. Thus, grazing-mortality counterbalanced growth, keeping V. cholerae populations in check. Net population gains were observed under particularly intense bloom conditions when V. cholerae proliferated, overcoming grazing pressure terms in part via rapid growth (> 4 doublings day(-1)). Our results show V. cholerae is subject to protozoan control and capable of utilizing multiple proliferation pathways in the marine environment. These findings suggest food web effects play a significant role controlling this pathogen's proliferation in coastal waters and should be considered in predictive models of disease risk. PMID- 16343319 TI - Growth of nutrient-replete Microcystis PCC 7806 cultures is inhibited by an extracellular signal produced by chlorotic cultures. AB - The frequency of cyanobacterial blooms has been increasing all over the world. These blooms are often toxic and have become a serious health problem. The aim of this work was to search for population density control mechanisms that could inhibit the proliferation of the toxic bloom-forming genus Microcystis. Microcystis PCC 7806 cultured for long periods in liquid ASM-1 medium loses its characteristic green colour. When a medium of chlorotic cultures is added to a nutrient-replete culture, cell density increase is drastically reduced when compared with controls. Inhibition of cell proliferation occurs in Microcystis cultures from any growth stage and was not strain-specific, but other genera tested showed no response. Investigations on the mechanism of growth inhibition showed that cultures treated with the conditioned medium acquired a pale colour, with pigment concentration similar to that found in chlorotic cultures. Ultrastructural examination showed that the conditioned medium induced thylakoid membrane disorganization, typical of chlorotic cells, in nutrient-replete cultures. An active extract was obtained and investigations showed that activity was retained after heating and after addition of an apolar solvent. This indicates that activity of the conditioned medium from chlorotic cells results from non-protein, apolar compound(s). PMID- 16343320 TI - Geomicrobiological exploration and characterization of a novel deep-sea hydrothermal system at the TOTO caldera in the Mariana Volcanic Arc. AB - Novel hydrothermal activities accompanying effluent white smokers and elemental sulfur chimney structures at the north-east lava dome of the TOTO caldera depression in the Mariana Volcanic Arc have been explored and characterized by geochemical and microbiological surveys. White smoker hydrothermal fluids were observed in the potential hydrothermal activity centre of the field and represented the maximal temperature of 170 degrees C and the lowest pH of 1.6. The chimney structures, all consisting of elemental sulfur (sulfur chimney), were also unique to the TOTO caldera hydrothermal field. Microbial community structures in a sulfur chimney and its formation hydrothermal fluid with a high concentration of hydrogen sulfide (15 mM) have been investigated by culture dependent and -independent analyses. 16S rRNA gene clone analysis and fluorescence in situ hybridization (FISH) analysis revealed that epsilon Proteobacteria dominated the microbial communities in the sulfur chimney structure and formed a dense microbial mat covering the sulfur chimney surface. Archaeal phylotypes were consistently minor components in the communities and related to the genera Thermococcus, Pyrodictium, Aeropyrum, and the uncultivated archaeal group of 'deep-sea hydrothermal vent euryarchaeotal group'. Cultivation analysis suggested that the chemolithoautotrophs might play a significant ecological role as primary producers utilizing gas and sulfur compounds provided from hydrothermal fluids. PMID- 16343321 TI - Community and cultivation analysis of arsenite oxidizing biofilms at Hot Creek. AB - At Hot Creek in California, geothermally derived arsenite is rapidly oxidized to arsenate. This process is mediated by microorganisms colonizing the surfaces of submerged aquatic macrophytes in the creek. Here we describe a multifaceted approach to characterizing this biofilm community and its activity. Molecular techniques were used to describe the community as a function of 16S-rRNA gene diversity. Cultivation-based strategies were used to enumerate and isolate three novel arsenite oxidizers, strains YED1-18, YED6-4 and YED6-21. All three strains are beta-Proteobacteria, of the genus Hydrogenophaga. Because these strains were isolated from the highest (i.e. million-fold) dilutions of disrupted biofilm suspensions, they represent the most numerically significant arsenite oxidizers recovered from this community. One clone (Hot Creek Clone 44) obtained from an inventory of the 16S rDNA sequence diversity present in the biofilm was found to be 99.6% identical to the 16S rDNA sequence of the isolate YED6-21. On the basis of most probable number (MPN) analyses, arsenite-oxidizing bacteria were found to account for 6-56% of the cultivated members of the community. Using MPN values, we could estimate an upper bound on the value of V(max) for the community of 1 x 10(-9)micromole arsenite min(-1) cell(-1). This estimate represents the first normalization of arsenite oxidation rates to MPN cell densities for a microbial community in a field incubation experiment. PMID- 16343322 TI - Distribution and phylogeny of hexachlorocyclohexane-degrading bacteria in soils from Spain. AB - Hexachlorocyclohexane (HCH)-degrading bacteria are believed to mediate natural attenuation of HCH contamination and have potential for active bioremediation processes. This study addressed the very limited understanding of the distribution, diversity and substrate specificity of such bacteria from 13 soil samples, varying in levels of HCH contamination, from four sites in Spain. Hexachlorocyclohexane removal occurred in 16 of 36 enrichment cultures. Hexachlorocyclohexane-degrading populations were clearly associated with HCH contaminated soils, and populations growing on the delta-HCH isomer were only found in soil contaminated with delta-HCH. beta-Hexachlorocyclohexane was persistent in enrichment cultures, and there was no evidence for populations growing on beta-HCH. From alpha- and gamma-HCH enrichment cultures, nine HCH degrading isolates were obtained, which were all Sphingomonas spp. Attempts to isolate organisms from delta-HCH enrichment cultures failed. None of the isolates grew on HCH as a sole organic substrate in pure culture. All isolates degraded alpha- and gamma-HCH, and most degraded beta-HCH. delta-Hexachlorocyclohexane inhibited growth of most isolates, but could be degraded by cell suspensions of at least four strains. Denaturing gradient gel electrophoresis indicated that the isolates represented predominant populations in the enrichment cultures, but additional predominant populations, including some Pseudomonas spp., could not be isolated. PMID- 16343323 TI - Experimental simulation of transmission of an obligate aphid pathogen with aphid flight dispersal. AB - The wide dispersal of Entomophthorales-caused mycoses that usually regulate aphid populations is most likely to be associated with the flight of infected alates. This hypothesis was examined via simulated flight and postflight colonization of Myzus periscae alates exposed to spore showers of Pandora neoaphidis, a common obligate aphid pathogen prevalent world wide. A total number of 407 alates were showered in different batches, then individually flown in a computer-monitoring flight mill system and reared on cabbage leaves for 14-day free reproduction and contagious infection within progeny colonies at 20-23 degrees C. On average, 80.6% of them flew 2.6 km in 1-5 h, survived 3.2 days, produced 5.3 nymphs, and transmitted their infection to progeny successfully. However, 9.8% of the flown alates left no progeny although they survived at least 1 day prior to mycosis while the rest were not mycosed, producing significantly more nymphs during the first week. The flight distances of the infected (0.01-10.2 km) or uninfected alates (0.1-8.3 km) were exponentially correlated to the flight time (r( 2) >or= 0.98). When grouped by the flight ranges of <1.0, 1.0-3.0, 3.0-5.0 and >5.0 km, the number of live aphids and the proportion of mycosed individuals per progeny colony over colonization days fit well to a complex logistic model (r( 2) = 0.984) and modified Gompertz model (r( 2) = 0.978) respectively. Both models included flight distance, postflight survival time, premycosis fecundity and primary infection rate as independent variables to affect the developmental rates. The results highlight the significant role of infected alates in the wide dispersal of P. neoaphidis-caused mycoses among aphids. PMID- 16343324 TI - Identification of peptide metabolites of Microcystis (Cyanobacteria) that inhibit trypsin-like activity in planktonic herbivorous Daphnia (Cladocera). AB - Cyanobacteria are recognized as producers of a broad variety of bioactive metabolites. Among these, the peptides synthesized by the non-ribosomal peptide synthetase pathway occur in high structural variability. One class of cyanobacterial peptides, the cyanopeptolins or micropeptins, have been shown to be strong inhibitors of vertebrate serine proteases, like trypsin. In the present study we screened extracts of ten strains of the unicellular cyanobacterium Microcystis sp. for their potential to inhibit trypsin-like activity in the planktonic crustacea Daphnia, the main herbivores in freshwater ecosystem. Respective standardized IC(50)'s varied for nearly two orders of magnitude. In HPLC fractions we could identify mainly cyanopeptolins as active compounds by MALDI-TOF mass spectrometry. Cyanopeptolins were found in 22 structural variants with 13 variants produced by one strain alone. Peptides of the microviridin class were moderately active while no activity was evident for microginins and microcystins. Among the cyanopeptolins only those were active that had an arginine or lysine residue N-terminal to the modified amino acid 3-amino-6 hydroxy-piperidone. Structural variants that had a tyrosine residue at this particular position did not inhibit trypsin-like activity. The highly variable composition of the side chain of cyanopeptolins had no marked effect on the activity. Among the six cyanobacterial strains we tested intensively two did not produce any cyanopeptolins and were accordingly less active as crude extracts. The present study underlines the potential importance of the biochemistry of cyanobacteria for the feeding ecology of a planktonic herbivore. PMID- 16343325 TI - Microbial life in ridge flank crustal fluids. AB - To determine the microbial community diversity within old oceanic crust, a novel sampling strategy was used to collect crustal fluids at Baby Bare Seamount, a 3.5 Ma old outcrop located in the north-east Pacific Ocean on the eastern flank of the Juan de Fuca Ridge. Stainless steel probes were driven directly into the igneous ocean crust to obtain samples of ridge flank crustal fluids. Genetic signatures and enrichment cultures of microorganisms demonstrate that these crustal fluids host a microbial community composed of species indigenous to the subseafloor, including anaerobic thermophiles, and species from other deep-sea habitats, such as seawater and sediments. Evidence using molecular techniques indicates the presence of a relatively small but active microbial population, dominated by bacteria. The microbial community diversity found in the crustal fluids may indicate habitat variability in old oceanic crust, with inputs of nutrients from seawater, sediment pore-water fluids and possibly hydrothermal sources. This report further supports the presence of an indigenous microbial community in ridge flank crustal fluids and advances our understanding of the potential physiological and phylogenetic diversity of this community. PMID- 16343326 TI - Anaerobic redox cycling of iron by freshwater sediment microorganisms. AB - The potential for microbially mediated anaerobic redox cycling of iron (Fe) was examined in a first-generation enrichment culture of freshwater wetland sediment microorganisms. Most probable number enumerations revealed the presence of significant populations of Fe(III)-reducing (approximately 10(8) cells ml(-1)) and Fe(II)-oxidizing, nitrate-reducing organisms (approximately 10(5) cells ml( 1)) in the freshwater sediment used to inoculate the enrichment cultures. Nitrate reduction commenced immediately following inoculation of acetate-containing (approximately 1 mM) medium with a small quantity (1% v/v) of wetland sediment, and resulted in the transient accumulation of NO(2)(-) and production of a mixture of gaseous end-products (N(2)O and N(2)) and NH(4)(+). Fe(III) oxide (high surface area goethite) reduction took place after NO(3)(-) was depleted and continued until all the acetate was utilized. Addition of NO(3)(-) after Fe(III) reduction ceased resulted in the immediate oxidation of Fe(II) coupled to reduction of NO(3)(-) to NH(4)(+). No significant NO(2)(-) accumulation was observed during nitrate-dependent Fe(II) oxidation. No Fe(II) oxidation occurred in pasteurized controls. Microbial community structure in the enrichment was monitored by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified 16S rDNA and reverse transcription polymerase chain reaction amplified 16S rRNA, as well as by construction of 16S rDNA clone libraries for four different time points during the experiment. Strong similarities in dominant members of the microbial community were observed in the Fe(III) reduction and nitrate-dependent Fe(II) oxidation phases of the experiment, specifically the common presence of organisms closely related (>or= 95% sequence similarity) to the genera Geobacter and Dechloromonas. These results indicate that the wetland sediments contained organisms such as Geobacter sp. which are capable of both dissimilatory Fe(III) reduction and oxidation of Fe(II) with reduction of NO(3)( ) to NH(4)(+). Our findings suggest that microbially catalysed nitrate-dependent Fe(II) oxidation has the potential to contribute to a dynamic anaerobic Fe redox cycle in freshwater sediments. PMID- 16343327 TI - Colonization of nascent, deep-sea hydrothermal vents by a novel Archaeal and Nanoarchaeal assemblage. AB - Active deep-sea hydrothermal vents are areas of intense mixing and severe thermal and chemical gradients, fostering a biotope rich in novel hyperthermophilic microorganisms and metabolic pathways. The goal of this study was to identify the earliest archaeal colonizers of nascent hydrothermal chimneys, organisms that may be previously uncharacterized as they are quickly replaced by a more stable climax community. During expeditions in 2001 and 2002 to the hydrothermal vents of the East Pacific Rise (EPR) (9 degrees 50'N, 104 degrees 17'W), we removed actively venting chimneys and in their place deployed mineral chambers and sampling units that promoted the growth of new, natural hydrothermal chimneys and allowed their collection within hours of formation. These samples were compared with those collected from established hydrothermal chimneys from EPR and Guaymas Basin vent sites. Using molecular and phylogenetic analysis of the 16S rDNA, we show here that at high temperatures, early colonization of a natural chimney is dominated by members of the archaeal genus Ignicoccus and its symbiont, Nanoarchaeum. We have identified 19 unique sequences closely related to the nanoarchaeal group, and five archaeal sequences that group closely with Ignicoccus. These organisms were found to colonize a natural, high temperature protochimney and vent-like mineral assemblages deployed over high temperature outflows within 92 h. When compared phylogenetically, several of these colonizing organisms form a unique clade independent of those found in mature chimneys and low-temperature mineral chamber samples. As a model ecosystem, the identification of pioneering consortia in deep-sea hydrothermal vents may help advance the understanding of how early microbial life forms gained a foothold in hydrothermal systems on early Earth and potentially on other planetary bodies. PMID- 16343328 TI - Composition of microbial communities in hexachlorocyclohexane (HCH) contaminated soils from Spain revealed with a habitat-specific microarray. AB - Microarray technology was used to characterize and compare hexachlorocyclohexane (HCH) contaminated soils from Spain. A library of 2,290 hypervariable 16S rRNA gene sequences was prepared with serial analysis of ribosomal sequence tags (SARST) from a composite of contaminated and uncontaminated soils. By designing hybridization probes specific to the 100 most abundant ribosomal sequence tags (RSTs) in the composite library, the RST array was designed to be habitat specific and predicted to monitor the most abundant polymerase chain reaction (PCR)-amplified phylotypes in the individual samples. The sensitivity and specificity of the RST array was tested with a series of pure culture-specific probes and hybridized with labelled soil PCR products to generate hybridization patterns for each soil. Sequencing of prominent bands in denaturing gradient gel electrophoresis (DGGE) fingerprints derived from these soils provided a means by which we successfully confirmed the habitat-specific array design and validated the bulk of the probe signals. Non-metric multidimensional scaling revealed correlations between probe signals and soil physicochemical parameters. Among the strongest correlations to total HCH contamination were probe signals corresponding to unknown Gamma Proteobacteria, potential pollutant-degrading phylotypes, and several organisms with acid-tolerant phenotypes. The strongest correlations to alpha-HCH were probe signals corresponding to the genus Sphingomonas, which contains known HCH degraders. This suggests that the population detected was enriched in situ by HCH contamination and may play a role in HCH degradation. Other environmental parameters were also likely instrumental in shaping community composition in these soils. The results highlight the power of habitat-specific microarrays for comparing complex microbial communities. PMID- 16343329 TI - Molecular analysis of deep subsurface Cretaceous rock indicates abundant Fe(III)- and S(zero)-reducing bacteria in a sulfate-rich environment. AB - A multilevel sampler (MLS) was emplaced in a borehole straddling anaerobic, sulfate-rich Cretaceous-era shale and sandstone rock formations approximately 200 m below ground surface at Cerro Negro, New Mexico. Sterile quartzite sand contained in chambers in the sampler allowed in situ colonization and recovery of nucleic acids for molecular analyses. Denaturing gradient gel electrophoresis and 16S rRNA gene cloning results indicated a homogeneously distributed bacterial community across the shale-sandstone interface. delta-Proteobacteria sequences were common at all depths, and were dominated by members of the Geobacteraceae family (Pelobacter, Desulphuromonas and Geobacter). Other members of this group are capable of dissimilatory Fe(III) and/or S degrees reduction, but not sulfate reduction. RNA hybridization data also suggested that Fe(III)-/S degrees reducing bacteria were predominant. These findings are striking considering the lack of significant concentrations of these electron acceptors in this environment. The next most abundant bacterial group indicated was the sulfate reducers, including Desulfobacterium, Desulfocapsa and Desulfobulbus. Sequences related to fermenters, denitrifiers and acetogens were also recovered. The presence of a phylogenetically and functionally diverse microbial community in this deep subsurface environment likely reflects the complex nature of the primary energy and carbon sources, kerogen associated with the shale. PMID- 16343330 TI - Stable carbon isotope fractionation during degradation of dichloromethane by methylotrophic bacteria. AB - Stable carbon isotope fractionation during dichloromethane (DCM) degradation by methylotrophic bacteria was investigated under aerobic and nitrate-reducing conditions. The strains studied comprise several Hyphomicrobium strains, Methylobacterium, Methylopila, Methylophilus and Methylorhabdus spp. that are considered to degrade DCM by a glutathione (GSH)-dependent dehalogenase enzyme system in the initial step. The stable carbon isotope fractionation factors (alphaC) of the strains varied under aerobic conditions between 1.043 and 1.071 and under nitrate-reducing conditions between 1.048 and 1.065. Comparison of isotope fractionation under aerobic and nitrate-reducing conditions by individual strains revealed only minor to no differences. The variability in isotope fractionation among strains was found to be related to the polymorphism of the functional genes encoding the DCM dehalogenase. PMID- 16343331 TI - Growth phase-dependent expression of the Pseudomonas putida KT2440 transcriptional machinery analysed with a genome-wide DNA microarray. AB - Bacterial transcriptional networks are built on a hierarchy of regulators, on top of which lie the components of the RNA polymerase (in particular the sigma factors) and the global control elements, which play a pivotal role. We have designed a genome-wide oligonucleotide-based DNA microarray for Pseudomonas putida KT2440. In combination with real-time reverse transcription polymerase chain reaction (RT-PCR), we have used it to analyse the expression pattern of the genes encoding the RNA polymerase subunits (the core enzyme and the 24 sigma factors), and various proteins involved in global regulation (Crc, Lrp, Fur, Anr, Fis, CsrA, IHF, HupA, HupB, HupN, BipA and several MvaT-like proteins), during the shift from exponential growth in rich medium into starvation and stress brought about by the entry into stationary phase. Expression of the genes encoding the RNA polymerase core and the vegetative sigma factor decreased in stationary phase, while that of sigma(S) increased. Data obtained for sigma(N), sigma(H), FliA and for the 19 extracytoplasmic function (ECF)-like sigma factors suggested that their mRNA levels change little upon entry into stationary phase. Expression of Crc, BipA, Fis, HupB, HupN and the MvaT-like protein PP3693 decreased in stationary phase, while that of HupA and the MvaT-like protein PP3765 increased significantly. Expression of IHF was indicative of post transcriptional control. These results provide the first global study of the expression of the transcriptional machinery through the exponential stationary phase shift in P. putida. PMID- 16343332 TI - Isolation and characterization of Pseudoalteromonas strains with divergent polycyclic aromatic hydrocarbon catabolic properties. AB - Fifteen strains of polycyclic aromatic hydrocarbon (PAH)-catabolizing bacteria, identified as Pseudoalteromonas spp. were isolated from Eagle Harbor, Puget Sound, USA, using a most probable number procedure in which naphthalene or phenanthrene was the sole carbon and energy source. Despite having identical 16S rDNA sequences, some catabolized many PAHs, whereas others oxidized only naphthalenes. A putative naphthalene 1,2-dioxygenase gene fragment was polymerase chain reaction-amplified from the naphthalene-degrading strains and shown to be almost identical to a gene present in Neptunomonas naphthovorans, suggesting horizontal transfer. PMID- 16343334 TI - InterferenceAnalyzer: tools for the analysis and simulation of multi-locus genetic data. AB - BACKGROUND: Good statistical models for analyzing and simulating multilocus recombination data exist but are not accessible to many biologists because their use requires reasonably sophisticated mathematical and computational implementation. While some labs have direct access to statisticians or programmers competent to carry out such analyses, many labs do not. We have created a platform independent application with an easy-to-use graphical user interface that will carry out such analyses including the simulations needed to bootstrap confidence intervals for the parameters of interest. This software should make multi-locus techniques accessible to labs that previously relied on less powerful and potentially statistically confounded single interval or double interval techniques. RESULTS: We introduce InterferenceAnalyzer, an implementation with a user-friendly graphical interface incorporating previously developed algorithms for the analysis and simulation of multilocus recombination data. We demonstrate the use and features of the program with an example of multilocus tetrad data from the mustard plant, Arabidopsis thaliana, and the yeast, Saccharomyces cerevisiae. CONCLUSION: InterferenceAnalyzer provides easy access to the powerful and appropriate statistical tools for the multi-locus analysis of genetic data. PMID- 16343335 TI - Copine A, a calcium-dependent membrane-binding protein, transiently localizes to the plasma membrane and intracellular vacuoles in Dictyostelium. AB - BACKGROUND: Copines are soluble, calcium-dependent membrane binding proteins found in a variety of organisms. Copines are characterized as having two C2 domains at the N-terminal region followed by an "A domain" at the C-terminal region. The "A domain" is similar in sequence to the von Willebrand A (VWA) domain found in integrins. The presence of C2 domains suggests that copines may be involved in cell signaling and/or membrane trafficking pathways. RESULTS: We have identified six copines genes in the Dictyostelium discoideum genome, cpnA cpnF, and have focused our studies on cpnA. CpnA is expressed throughout development and was shown to be capable of binding to membranes in a calcium dependent manner in vitro. A GFP-tagged CpnA was also capable of binding to membranes in a calcium-dependent manner in vitro. In live wildtype Dictyostelium cells expressing GFP-CpnA, GFP-CpnA was typically found in the cytoplasm without any specific localization to membranes. However, in a small subset of starved cells, GFP-CpnA was observed to bind transiently (typically approximately 1-10 s) to the plasma membrane and intracellular vacuoles. In some cells, the transient membrane localization of GFP-CpnA was observed to occur multiple times in an oscillatory manner over several minutes. In plasma membrane disrupted cells, GFP CpnA was observed to associate with the plasma membrane and intracellular vacuoles in a calcium-dependent manner. In fixed cells, GFP-CpnA labeled the plasma membrane and intracellular vacuoles, including contractile vacuoles, organelles of the endolysosomal pathway, and phagosomes. CONCLUSION: Our results show that Dictyostelium has multiple copine homologs and provides an excellent system in which to study copine function. The localization of a GFP-tagged CpnA to the plasma membrane, contractile vacuoles, organelles of the endolysosomal pathway, and phagosomes suggests that CpnA may have a role in the function of these organelles or the trafficking to and from them. In addition, we hypothesize that the observed transient oscillatory membrane localization of GFP-CpnA in a small subset of starved cells is caused by fast calcium waves and speculate that CpnA may have a role in development, particularly in the differentiation of stalk cells. PMID- 16343336 TI - (Cost)-effectiveness of case-management by district nurses among primary informal caregivers of older adults with dementia symptoms and the older adults who receive informal care: design of a randomized controlled trial [ISCRTN83135728]. AB - BACKGROUND: Dementia is an incurable disease with devastating consequences for both patients and their relatives. The objective of this study is to describe the study protocol of a randomized controlled trial with assignment to either usual care or case-management by district nurses, among informal caregivers of older adults with dementia symptoms who live at home and the older adults who receive informal care. METHODS/DESIGN: In this randomized controlled trial, effectiveness as well as cost-effectiveness of case-management is evaluated. It concerns case management in early-detected patients with dementia symptoms and their primary informal caregivers. Participants are followed up to twelve months after baseline assessment. The main outcome measure of the effect evaluation is the caregiver's sense of competence to care for the older person with dementia symptoms. The economic evaluation is performed from a societal perspective. DISCUSSION: This is one of the first trials on case-management that includes an economic evaluation. In addition, it concerns a tailor-made intervention in early-detected patients with dementia symptoms and their caregivers. The results of this randomized controlled trial will provide valuable information for health professionals and policy makers on effectiveness and cost-effectiveness of early tailor-made case management for patients and their informal caregivers. Moreover, positive effects will challenge current health care systems to move to more pro-active approaches for this group. PMID- 16343337 TI - Kalign--an accurate and fast multiple sequence alignment algorithm. AB - BACKGROUND: The alignment of multiple protein sequences is a fundamental step in the analysis of biological data. It has traditionally been applied to analyzing protein families for conserved motifs, phylogeny, structural properties, and to improve sensitivity in homology searching. The availability of complete genome sequences has increased the demands on multiple sequence alignment (MSA) programs. Current MSA methods suffer from being either too inaccurate or too computationally expensive to be applied effectively in large-scale comparative genomics. RESULTS: We developed Kalign, a method employing the Wu-Manber string matching algorithm, to improve both the accuracy and speed of multiple sequence alignment. We compared the speed and accuracy of Kalign to other popular methods using Balibase, Prefab, and a new large test set. Kalign was as accurate as the best other methods on small alignments, but significantly more accurate when aligning large and distantly related sets of sequences. In our comparisons, Kalign was about 10 times faster than ClustalW and, depending on the alignment size, up to 50 times faster than popular iterative methods. CONCLUSION: Kalign is a fast and robust alignment method. It is especially well suited for the increasingly important task of aligning large numbers of sequences. PMID- 16343338 TI - Flecainide overdose--support using an intra-aortic balloon pump. AB - BACKGROUND: Flecainide is an antiarrhythmic agent which is being used increasingly for the management of super-ventricular arrhythmias. Overdose with flecainide is frequently fatal with mortality reported as high as 22% due to arrhythmias, myocardial depression and conduction defects leading to electro mechanical dissociation and asytole. Supportive measures are often required during the case and previously have included inotropes, extracorporeal membrane oxygenation and cardiopulmonary bypass. CASE PRESENTATION: A 47 year old lady presented to the emergency department with a four hour history of severe central chest pain. Her ECG showed atrial fibrillation and broad QRS complexes with a sine wave appearance. She had a past history of paroxysmal atrial fibrillation and significant psychiatric history. Following thrombolysis for a presumed myocardial infarction she developed cardiogenic shock with severely impaired left ventricular function. An intra-aortic balloon pump was inserted and coronary angiography demonstrated normal coronary arteries. With inotropic support she improved over 48 hours, with both her QRS duration and left ventricular function returning to normal. Biochemical testing following her discharge demonstrated significantly elevated levels of flecainide. CONCLUSION: The use of an intra aortic balloon pump is a useful supportive measure during the acute phase of flecainide overdose associated with severe myocardial depression. PMID- 16343339 TI - National estimates for maternal mortality: an analysis based on the WHO systematic review of maternal mortality and morbidity. AB - BACKGROUND: Despite the worldwide commitment to improving maternal health, measuring, monitoring and comparing maternal mortality estimates remain a challenge. Due to lack of data, international agencies have to rely on mathematical models to assess its global burden. In order to assist in mapping the burden of reproductive ill-health, we conducted a systematic review of incidence/prevalence of maternal mortality and morbidity. METHODS: We followed the standard methodology for systematic reviews. This manuscript presents nationally representative estimates of maternal mortality derived from the systematic review. Using regression models, relationships between study-specific and country-specific variables with the maternal mortality estimates are explored in order to assist further modelling to predict maternal mortality. RESULTS: Maternal mortality estimates included 141 countries and represent 78.1% of the live births worldwide. As expected, large variability between countries, and within regions and subregions, is identified. Analysis of variability according to study characteristics did not yield useful results given the high correlation with each other, with development status and region. A regression model including selected country-specific variables was able to explain 90% of the variability of the maternal mortality estimates. Among all country-specific variables selected for the analysis, three had the strongest relationships with maternal mortality: proportion of deliveries assisted by a skilled birth attendant, infant mortality rate and health expenditure per capita. CONCLUSION: With the exception of developed countries, variability of national maternal mortality estimates is large even within subregions. It seems more appropriate to study such variation through differentials in other national and subnational characteristics. Other than region, study of country-specific variables suggests infant mortality rate, skilled birth attendant at delivery and health expenditure per capita are key variables to predict maternal mortality at national level. PMID- 16343340 TI - The role and performance of chest X-ray for the diagnosis of tuberculosis: a cost effectiveness analysis in Nairobi, Kenya. AB - BACKGROUND: The objective of this study was to establish 1) the performance of chest X-ray (CXR) in all suspects of tuberculosis (TB), as well as smear-negative TB suspects and 2) to compare the cost-effectiveness of the routine diagnostic pathway using Ziehl-Neelsen (ZN) sputum microscopy followed by CXR if case of negative sputum result (ZN followed by CXR) with an alternative pathway using CXR as a screening tool (CXR followed by ZN). METHODS: From TB suspects attending a chest clinic in Nairobi, Kenya, three sputum specimens were examined for ZN and culture (Lowenstein Jensen). Culture was used as gold standard. From each suspect a CXR was made using a four point scoring system: i: no pathology, ii: pathology not consistent for TB, iii: pathology consistent for TB and iv: pathology highly consistent for TB. The combined score i + ii was labeled as "no TB" and the combined score iii + iv was labeled as "TB". Films were re-read by a reference radiologist. HIV test was performed on those who consented. Laboratory and CXR costs were used to compare for cost-effectiveness. RESULTS: Of the 1,389 suspects enrolled, for 998 (72%) data on smear, culture and CXR was complete. 714 films were re-read, showing a 89% agreement (kappa value = 0.75 s.e.0.037) for the combined scores "TB" or "no-TB". The sensitivity/specificity of the CXR score "TB" among smear-negative suspects was 80%/67%. Using chest CXR as a screening tool in all suspects, sensitivity/specificity of the score "any pathology" was 92%, respectively 63%. The cost per correctly diagnosed case was for the routine process 8.72 dollars, compared to 9.27 dollars using CXR as screening tool. When costs of treatment were included, CXR followed by ZN became more cost-effective. CONCLUSION: The diagnostic pathway ZN followed by CXR was more cost-effective as compared to CXR followed by ZN. When cost of treatment was also considered CXR followed by ZN became more cost-effective. The low specificity of chest X-ray remains a subject of concern. Depending whether CXR was performed on all suspects or on smear-negative suspects only, 22%-45% of patients labeled as "TB" had a negative culture. The introduction of a well-defined scoring system, clinical conferences and a system of CXR quality control can contribute to improved diagnostic performance. PMID- 16343341 TI - The course of mental health after miscarriage and induced abortion: a longitudinal, five-year follow-up study. AB - BACKGROUND: Miscarriage and induced abortion are life events that can potentially cause mental distress. The objective of this study was to determine whether there are differences in the patterns of normalization of mental health scores after these two pregnancy termination events. METHODS: Forty women who experienced miscarriages and 80 women who underwent abortions at the main hospital of Buskerud County in Norway were interviewed. All subjects completed the following questionnaires 10 days (T1), six months (T2), two years (T3) and five years (T4) after the pregnancy termination: Impact of Event Scale (IES), Quality of Life, Hospital Anxiety and Depression Scale (HADS), and another addressing their feelings about the pregnancy termination. Differential changes in mean scores were determined by analysis of covariance (ANCOVA) and inter-group differences were assessed by ordinary least squares methods. RESULTS: Women who had experienced a miscarriage had more mental distress at 10 days and six months after the pregnancy termination than women who had undergone an abortion. However, women who had had a miscarriage exhibited significantly quicker improvement on IES scores for avoidance, grief, loss, guilt and anger throughout the observation period. Women who experienced induced abortion had significantly greater IES scores for avoidance and for the feelings of guilt, shame and relief than the miscarriage group at two and five years after the pregnancy termination (IES avoidance means: 3.2 vs 9.3 at T3, respectively, p < 0.001; 1.5 vs 8.3 at T4, respectively, p < 0.001). Compared with the general population, women who had undergone induced abortion had significantly higher HADS anxiety scores at all four interviews (p < 0.01 to p < 0.001), while women who had had a miscarriage had significantly higher anxiety scores only at T1 (p < 0.01). CONCLUSION: The course of psychological responses to miscarriage and abortion differed during the five-year period after the event. Women who had undergone an abortion exhibited higher scores during the follow-up period for some outcomes. The difference in the courses of responses may partly result from the different characteristics of the two pregnancy termination events. PMID- 16343342 TI - Socioeconomic status and non-fatal injuries among Canadian adolescents: variations across SES and injury measures. AB - BACKGROUND: While research to date has consistently demonstrated that socioeconomic status (SES) is inversely associated with injury mortality in both children and adults, findings have been less consistent for non-fatal injuries. The literature addressing SES and injury morbidity among adolescents has been particularly inconclusive. To explore potential explanations for these discrepant research findings, this study uniquely compared the relationship across different measures of SES and different causes of injury (recreation versus non-recreation injuries) within a sample of Canadian adolescents. METHODS: The sample included adolescent participants (aged 12 to 19 years) in the Canadian 1996-1997 cross sectional National Population Health Survey (n = 6967). Five SES measures (household income, two neighbourhood-level proxy measures, two parental indicators) were examined in relation to three injury outcomes (total, recreation, and non-recreation injuries) using multivariable logistic regression. RESULTS: Among males, a clear relationship with injury was observed only for a parental SES index, which was positively associated with total and recreation injuries (odds ratios for the highest versus lowest SES category of 1.9 for total and 2.5 for recreation injuries). Among females, there was some evidence of a positive relationship between SES and injuries, particularly for a neighbourhood level education measure with total and recreation injuries (odds ratios of 1.7 for total and 2.0 for recreation injuries). CONCLUSION: The results suggest that differences related to the measures of SES chosen and the causes of injury under study may both contribute to discrepancies in past research on SES and non-fatal injuries among adolescents. To clarify the potential SES-injury relationship among youth, the findings emphasize a need for a greater understanding of the meaning and relevance of different SES measures for adolescents, and for an exploration of the pathways through which SES may be related to injury risk. PMID- 16343343 TI - Association between frequent use of nonsteroidal anti-inflammatory drugs and breast cancer. AB - BACKGROUND: Eighty percent of all breast cancers and almost 90% of breast cancer deaths occur among post-menopausal women. We used a nested case control design to examine the association between nonsteroidal anti-inflammatory drug (NSAID) use and breast cancer occurrence among women over 65 years of age. The cyclooxygenase (COX)-2 enzyme is expressed more in breast cancers than in normal breast tissue. COX-2 inhibition may have a role in breast cancer prevention. METHODS: In the Canadian province of Quebec, physician services are covered through a governmental insurance plan. Medication costs are covered for those > or = 65 years of age and a publicly funded screening program for breast cancer targets all women 50 years of age or older. We obtained encrypted data from these insurance databases on all women > or = 65 years of age who filled a prescription for COX-2 inhibitors, non-selective NSAIDs (ns-NSAIDs), aspirin, or acetaminophen between January 1998 and December 2002. Cases were defined as those women who have undergone mammography between April 2001 and June 2002 and had a diagnosis of breast cancer within six months following mammography. Controls included those who have undergone mammography between April 2001 and June 2002 without a diagnosis of any cancer during the six months following mammography. The exposure of interest, frequent NSAID use, was defined as use of ns-NSAIDs and/or COX-2 inhibitors for > or = 90 days during the year prior to mammography. Frequent use served as a convenient proxy for long term chronic use. RESULTS: We identified 1,090 cases and 44,990 controls. Cases were older and more likely to have breast cancer risk factors. Logistic regression models adjusting for potential confounders showed that frequent use of ns-NSAIDs and/or COX-2 inhibitors was associated with a lower risk of breast cancer (OR: 0.75, 95% confidence interval 0.64-0.89). Results were similar for COX-2 inhibitors (0.81, 0.68-0.97) and ns NSAIDs (0.65, 0.43-0.99), when assessed separately. Frequent use of aspirin at doses > 100 mg/day in the year prior to mammography was also associated with a lower risk of breast cancer (0.75, 0.64-0.89). However, use of aspirin at doses < or = 100 mg/day did not have any association with breast cancer (0.91, 0.71 1.16). CONCLUSION: Women who use NSAIDs or doses of aspirin > 100 mg frequently may have a lower risk of breast cancer. PMID- 16343344 TI - Dual-modality impairment of implicit learning of letter-strings versus color patterns in patients with schizophrenia. AB - BACKGROUND: Implicit learning was reported to be intact in schizophrenia using artificial grammar learning. However, emerging evidence indicates that artificial grammar learning is not a unitary process. The authors used dual coding stimuli and schizophrenia clinical symptom dimensions to re-evaluate the effect of schizophrenia on various components of artificial grammar learning. METHODS: Letter string and color pattern artificial grammar learning performances were compared between 63 schizophrenic patients and 27 comparison subjects. Four symptom dimensions derived from a Chinese Positive and Negative Symptom Scale ratings were correlated with patients' artificial grammar implicit learning performances along the two stimulus dimensions. Patients' explicit memory performances were assessed by verbal paired associates and visual reproduction subtests of the Wechsler Memory Scales Revised Version to provide a contrast to their implicit memory function. RESULTS: Schizophrenia severely hindered color pattern artificial grammar learning while the disease affected lexical string artificial grammar learning to a lesser degree after correcting the influences from age, education and the performance of explicit memory function of both verbal and visual modalities. Both learning performances correlated significantly with the severity of patients' schizophrenic clinical symptom dimensions that reflect poor abstract thinking, disorganized thinking, and stereotyped thinking. CONCLUSION: The results of this study suggested that schizophrenia affects various mechanisms of artificial grammar learning differently. Implicit learning, knowledge acquisition in the absence of conscious awareness, is not entirely intact in patients with schizophrenia. Schizophrenia affects implicit learning through an impairment of the ability of making abstractions from rules and at least in part decreasing the capacity for perceptual learning. PMID- 16343345 TI - Cost, affordability and cost-effectiveness of strategies to control tuberculosis in countries with high HIV prevalence. AB - BACKGROUND: The HIV epidemic has caused a dramatic increase in tuberculosis (TB) in East and southern Africa. Several strategies have the potential to reduce the burden of TB in high HIV prevalence settings, and cost and cost-effectiveness analyses can help to prioritize them when budget constraints exist. However, published cost and cost-effectiveness studies are limited. METHODS: Our objective was to compare the cost, affordability and cost-effectiveness of seven strategies for reducing the burden of TB in countries with high HIV prevalence. A compartmental difference equation model of TB and HIV and recent cost data were used to assess the costs (year 2003 USD prices) and effects (TB cases averted, deaths averted, DALYs gained) of these strategies in Kenya during the period 2004 2023. RESULTS: The three lowest cost and most cost-effective strategies were improving TB cure rates, improving TB case detection rates, and improving both together. The incremental cost of combined improvements to case detection and cure was below USD 15 million per year (7.5% of year 2000 government health expenditure); the mean cost per DALY gained of these three strategies ranged from USD 18 to USD 34. Antiretroviral therapy (ART) had the highest incremental costs, which by 2007 could be as large as total government health expenditures in year 2000. ART could also gain more DALYs than the other strategies, at a cost per DALY gained of around USD 260 to USD 530. Both the costs and effects of treatment for latent tuberculosis infection (TLTI) for HIV+ individuals were low; the cost per DALY gained ranged from about USD 85 to USD 370. Averting one HIV infection for less than USD 250 would be as cost-effective as improving TB case detection and cure rates to WHO target levels. CONCLUSION: To reduce the burden of TB in high HIV prevalence settings, the immediate goal should be to increase TB case detection rates and, to the extent possible, improve TB cure rates, preferably in combination. Realising the full potential of ART will require substantial new funding and strengthening of health system capacity so that increased funding can be used effectively. PMID- 16343346 TI - GPX-Macrophage Expression Atlas: a database for expression profiles of macrophages challenged with a variety of pro-inflammatory, anti-inflammatory, benign and pathogen insults. AB - BACKGROUND: Macrophages play an integral role in the host immune system, bridging innate and adaptive immunity. As such, they are finely attuned to extracellular and intracellular stimuli and respond by rapidly initiating multiple signalling cascades with diverse effector functions. The macrophage cell is therefore an experimentally and clinically amenable biological system for the mapping of biological pathways. The goal of the macrophage expression atlas is to systematically investigate the pathway biology and interaction network of macrophages challenged with a variety of insults, in particular via infection and activation with key inflammatory mediators. As an important first step towards this we present a single searchable database resource containing high-throughput macrophage gene expression studies. DESCRIPTION: The GPX Macrophage Expression Atlas (GPX-MEA) is an online resource for gene expression based studies of a range of macrophage cell types following treatment with pathogens and immune modulators. GPX-MEA follows the MIAME standard and includes an objective quality score with each experiment. It places special emphasis on rigorously capturing the experimental design and enables the searching of expression data from different microarray experiments. Studies may be queried on the basis of experimental parameters, sample information and quality assessment score. The ability to compare the expression values of individual genes across multiple experiments is provided. In addition, the database offers access to experimental annotation and analysis files and includes experiments and raw data previously unavailable to the research community. CONCLUSION: GPX-MEA is the first example of a quality scored gene expression database focussed on a macrophage cellular system that allows efficient identification of transcriptional patterns. The resource will provide novel insights into the phenotypic response of macrophages to a variety of benign, inflammatory, and pathogen insults. GPX-MEA is available through the GPX website at http://www.gti.ed.ac.uk/GPX. PMID- 16343347 TI - Comparative genome analysis reveals a conserved family of actin-like proteins in apicomplexan parasites. AB - BACKGROUND: The phylum Apicomplexa is an early-branching eukaryotic lineage that contains a number of important human and animal pathogens. Their complex life cycles and unique cytoskeletal features distinguish them from other model eukaryotes. Apicomplexans rely on actin-based motility for cell invasion, yet the regulation of this system remains largely unknown. Consequently, we focused our efforts on identifying actin-related proteins in the recently completed genomes of Toxoplasma gondii, Plasmodium spp., Cryptosporidium spp., and Theileria spp. RESULTS: Comparative genomic and phylogenetic studies of apicomplexan genomes reveals that most contain only a single conventional actin and yet they each have 8-10 additional actin-related proteins. Among these are a highly conserved Arp1 protein (likely part of a conserved dynactin complex), and Arp4 and Arp6 homologues (subunits of the chromatin-remodeling machinery). In contrast, apicomplexans lack canonical Arp2 or Arp3 proteins, suggesting they lost the Arp2/3 actin polymerization complex on their evolutionary path towards intracellular parasitism. Seven of these actin-like proteins (ALPs) are novel to apicomplexans. They show no phylogenetic associations to the known Arp groups and likely serve functions specific to this important group of intracellular parasites. CONCLUSION: The large diversity of actin-like proteins in apicomplexans suggests that the actin protein family has diverged to fulfill various roles in the unique biology of intracellular parasites. Conserved Arps likely participate in vesicular transport and gene expression, while apicomplexan specific ALPs may control unique biological traits such as actin-based gliding motility. PMID- 16343348 TI - Monckeberg's sclerosis - is the artery the only target of calcification? AB - BACKGROUND: Since its first description, Monckeberg's sclerosis has only been related to arterial media calcification, being listed among the primary diseases of the vessels. CASE PRESENTATION: We report here a clinically and histologically confirmed case of Monckeberg's sclerosis in which the patient presented with massive areas of soft tissue calcifications in the pharynx and larynx. Polysomnographic parameters showed severe obstructive apnea refractory to nasal continuous positive airway pressure. Clinical and laboratory findings excluded concomitant endocrine or rheumatological diseases. CONCLUSION: Our data provide a new insight about Monckeberg's sclerosis, i.e., the fact that the etiopathogenic process involved in the phenomenon of calcification may not be restricted only to the arteries, but may occur in the entire organism. Further studies of the etiopathogenesis of this disease are needed. PMID- 16343349 TI - Stimulation of cannabinoid receptor 2 (CB2) suppresses microglial activation. AB - BACKGROUND: Activated microglial cells have been implicated in a number of neurodegenerative disorders, including Alzheimer's disease (AD), multiple sclerosis (MS), and HIV dementia. It is well known that inflammatory mediators such as nitric oxide (NO), cytokines, and chemokines play an important role in microglial cell-associated neuron cell damage. Our previous studies have shown that CD40 signaling is involved in pathological activation of microglial cells. Many data reveal that cannabinoids mediate suppression of inflammation in vitro and in vivo through stimulation of cannabinoid receptor 2 (CB2). METHODS: In this study, we investigated the effects of a cannabinoid agonist on CD40 expression and function by cultured microglial cells activated by IFN-gamma using RT-PCR, Western immunoblotting, flow cytometry, and anti-CB2 small interfering RNA (siRNA) analyses. Furthermore, we examined if the stimulation of CB2 could modulate the capacity of microglial cells to phagocytise Abeta1-42 peptide using a phagocytosis assay. RESULTS: We found that the selective stimulation of cannabinoid receptor CB2 by JWH-015 suppressed IFN-gamma-induced CD40 expression. In addition, this CB2 agonist markedly inhibited IFN-gamma-induced phosphorylation of JAK/STAT1. Further, this stimulation was also able to suppress microglial TNF-alpha and nitric oxide production induced either by IFN-gamma or Abeta peptide challenge in the presence of CD40 ligation. Finally, we showed that CB2 activation by JWH-015 markedly attenuated CD40-mediated inhibition of microglial phagocytosis of Abeta1-42 peptide. Taken together, these results provide mechanistic insight into beneficial effects provided by cannabinoid receptor CB2 modulation in neurodegenerative diseases, particularly AD. PMID- 16343350 TI - Diagnostic and treatment delay among pulmonary tuberculosis patients in Ethiopia: a cross sectional study. AB - BACKGROUND: Delayed diagnosis and treatment of tuberculosis (TB) results in severe disease and a higher mortality. It also leads to an increased period of infectivity in the community. The objective of this study was to determine the length of delays, and analyze the factors affecting the delay from onset of symptoms of pulmonary tuberculosis (PTB) until the commencement of treatment. METHODS: In randomly selected TB management units (TBMUs), i.e. government health institutions which have diagnosing and treatment facilities for TB in Amhara Region, we conducted a cross sectional study from September 1-December 31/2003. Delay was analyzed from two perspectives, 1. Period between onset of TB symptoms to first visit to any health provider (health seeking period), and from the first health provider visit to initiation of treatment (health providers' delay), and 2. Period between onset of TB symptoms to first visit to a medical provider (patients' delay), and from this visit to commencement of anti-TB treatment (health systems' delay). Patients were interviewed on the same date of diagnosis using a semi-structured questionnaire. Logistics regression analysis was applied to analyze the risk factors of delays. RESULTS: A total of 384 new smear positive PTB patients participated in the study. The median total delay was 80 days. The median health-seeking period and health providers' delays were 15 and 61 days, respectively. Conversely, the median patients' and health systems' delays were 30 and 21 days, respectively. Taking medical providers as a reference point, we found that forty eight percent of the subjects delayed for more than one month. Patients' delays were strongly associated with first visit to non-formal health providers and self treatment (P < 0.0001). Prior attendance to a health post/clinic was associated with increased health systems' delay (p < 0.0001). CONCLUSION: Delay in the diagnosis and treatment of PTB is unacceptably high in Amhara region. Health providers' and health systems' delays represent the major portion of the total delay. Accessing a simple and rapid diagnostic test for TB at the lowest level of health care facility and encouraging a dialogue among all health providers are imperative interventions. PMID- 16343351 TI - Stromal mesenchyme cell genes of the human prostate and bladder. AB - BACKGROUND: Stromal mesenchyme cells play an important role in epithelial differentiation and likely in cancer as well. Induction of epithelial differentiation is organ-specific, and the genes responsible could be identified through a comparative genomic analysis of the stromal cells from two different organs. These genes might be aberrantly expressed in cancer since cancer could be viewed as due to a defect in stromal signaling. We propose to identify the prostate stromal genes by analysis of differentially expressed genes between prostate and bladder stromal cells, and to examine their expression in prostate cancer. METHODS: Immunohistochemistry using antibodies to cluster designation (CD) cell surface antigens was first used to characterize the stromas of the prostate and bladder. Stromal cells were prepared from either prostate or bladder tissue for cell culture. RNA was isolated from the cultured cells and analyzed by DNA microarrays. Expression of candidate genes in normal prostate and prostate cancer was examined by RT-PCR. RESULTS: The bladder stroma was phenotypically different from that of the prostate. Most notable was the presence of a layer of CD13+ cells adjacent to the urothelium. This structural feature was also seen in the mouse bladder. The prostate stroma was uniformly CD13-. A number of differentially expressed genes between prostate and bladder stromal cells were identified. One prostate gene, proenkephalin (PENK), was of interest because it encodes a hormone. Secreted proteins such as hormones and bioactive peptides are known to mediate cell-cell signaling. Prostate stromal expression of PENK was verified by an antibody raised against a PENK peptide, by RT-PCR analysis of laser-capture microdissected stromal cells, and by database analysis. Gene expression analysis showed that PENK expression was down-regulated in prostate cancer. CONCLUSION: Our findings show that the histologically similar stromas of the prostate and bladder are phenotypically different, and express organ-specific genes. The importance of these genes in epithelial development is suggested by their abnormal expression in cancer. Among the candidates is the hormone PENK and the down-regulation of PENK expression in cancer suggests a possible association with cancer development. PMID- 16343352 TI - Repolarization of the action potential enabled by Na+ channel deactivation in PSpice simulation of cardiac muscle propagation. AB - BACKGROUND: In previous studies on propagation of simulated action potentials (APs) in cardiac muscle using PSpice modeling, we reported that a second black box (BB) could not be inserted into the K+ leg of the basic membrane unit because that caused the PSpice program to become very unstable. Therefore, only the rising phase of the APs could be simulated. This restriction was acceptable since only the mechanism of transmission of excitation from one cell to the next was being investigated. METHODS AND RESULTS: We have now been able to repolarize the AP by inserting a second BB into the Na+ leg of the basic units. This second BB effectively mimicked deactivation of the Na+ channel conductance. This produced repolarization of the AP, not by activation of K+ conductance, but by deactivation of the Na+ conductance. The propagation of complete APs was studied in a chain (strand) of 10 cardiac muscle cells, in which various numbers of gap junction (gj) channels (assumed to be 100 pS each) were inserted across the cell junctions. The shunt resistance across the junctions produced by the gj-channels (Rgj) was varied from 100,000 M? (0 gj-channels) to 10,000 M? (1 gj-channel), to 1,000 M? (10 channels), to 100 M? (100 channels), and 10 M? (1000 channels). The velocity of propagation (theta, in cm/s) was calculated from the measured total propagation time (TPT, the time difference between when the AP rising phase of the first cell and the last cell crossed -20 mV, assuming a cell length of 150 microm. When there were no gj-channels, or only a few, the transmission of excitation between cells was produced by the electric field (EF), i.e. the negative junctional cleft potential, that is generated in the narrow junctional clefts (e.g. 100 A) when the prejunctional membrane fires an AP. When there were many gj-channels (e.g. 1000 or 10,000), the transmission of excitation was produced by local-circuit current flow from one cell to the next through the gj channels. CONCLUSION: We have now been able to simulate complete APs in cardiac muscle cells that could propagate along a single chain of 10 cells, even when there were no gj-channels between the cells. PMID- 16343353 TI - High-resolution genotyping of Pseudomonas aeruginosa strains linked to acute post cataract surgery endophthalmitis outbreaks in India. AB - BACKGROUND: Investigation of two independent outbreaks of post cataract surgery endophthalmitis identified the reservoir of epidemic strains of P. aeruginosa. METHODS: Patient isolates cultured from vitreous fluid of all the nine cases and from the peripheral devices of phacoemulsification machine were subjected to high resolution Fluorescent Amplified Fragment Length Polymorphism (FAFLP) analysis. RESULTS: FAFLP based genotyping of the isolates confirmed nosocomial transmission. Although biochemical characterization and antibiotic susceptibility profiles grouped all the isolates together, FAFLP based genotyping revealed that, all the outbreak isolates were derived from 2 different strains, with independent origins. One group of isolates was traced to phacoprobe and the second one to the internal tubing system of the phacoemulsification machine used in cataract surgery. In silico analysis indicated possible evolution in both the clusters of P. aeruginosa isolates due to genetic polymorphisms. The polymorphisms were mapped to gene products (cell envelope, outer membrane proteins) possibly having significant role in pathogenesis. CONCLUSION: The present study is probably the first one to apply FAFLP typing successfully to investigate outbreaks of postoperative endophthalmitis (POE) in an ophthalmic setting, which was able to identify the source, and helped to make rational decisions on sterilization procedures that halted more cases of infection in these hospitals. PMID- 16343354 TI - Impact of standalone and embedded telephone triage systems on after hours primary medical care service utilisation and mix in Australia. AB - BACKGROUND: The Australian government sponsored five local trials aimed at addressing problems in after hours (AH) primary medical care (PMC). The study's objective was to determine if the four trials, where telephone triage was the sole innovation, led to a reduction in AH service utilisation and change in service mix towards AH GP clinics. Changes in utilisation and mix of AH GP clinic and home visits, ED and ambulance use were monitored in the trial areas, and in a national sample to adjust for the effects of secular trend. Pre- and post-trial telephone surveys of two separate random samples of approximately 350 AH PMC user households in each area were conducted. RESULTS: Some types of AH PMC use became more frequent in both of the standalone services using nurse-administered proprietary call centre software, which were aimed at better addressing population need (Statewide call centre; Regional call centre). Service use overall (95%CI: 1.03-1.83) and GP clinic use (95%CI: 1.07-2.00) increased in the metro area of the Statewide call centre and in GP clinic (95%CI: 1.04-2.14) and home visits (95%CI: 1.03-3.91) in the non-metro area of the Regional call centre. Service mix only changed in the non-metro area of the Regional call centre with increased contact in GP home visits (95%CI: 1.02-4.38). Levels of use remained unchanged in both embedded services using other than proprietary software, which were established to support the GP workforce (Deputising service; Local triage centre). Service mix only changed in the Deputising service with a change away from AH GP clinics in both contact (95%CI: 0.39-0.97) and frequency (95% CI: 2.12 - -0.7). CONCLUSION: Bearing in mind limitations in estimating AH PMC utilisation levels and mix, it is concluded that the impacts of telephone triage were generally smaller in Australia than reported elsewhere. There were different impacts on levels of service utilisation and service mix in standalone call centres and embedded services. Impacts of telephone triage on service utilisation and mix are influenced by the type of telephone triage offered, the goals of the agency providing the service, as well as local factors. PMID- 16343355 TI - A health policy for hearing impairment in older Australians: what should it include? AB - BACKGROUND: As in all western countries, Australia's older population experiences high levels of hearing impairment coupled with relatively low levels of hearing device usage. Poor hearing diminishes the quality of life of affected individuals and their families. This paper discusses how to improve Australian hearing health policies in order to better combat this impairment amongst older Australians. METHOD: We searched the databases Medline, Meditext and Web of Science to find articles that discussed strategies and innovations to assist the hearing health of older people, and related this material to observations made during the Blue Mountains Hearing Study in NSW between 1997 and 2003. RESULTS AND DISCUSSION: The literature search identified five areas for inclusion in a comprehensive hearing health policy in Australia. These are: early intervention; addressing of hearing aid expense; the use of assisted listening devices; hearing rehabilitation, and; screening and education. Further research in Australia is critical if we are to develop a strong approach to the increasing prevalence of age-related hearing loss. CONCLUSION: Australia needs to act now to address hearing impairment as it is a major cause of disability in those aged 55 and over. Federal and State governments should collaborate to construct a comprehensive hearing health policy that tackles poor levels of hearing health through early intervention, addressing hearing aid expense, encouraging the use of assisted listening devices, rehabilitation, screening and education. A good start would be to declare age related hearing impairment as a National Health Priority Area. PMID- 16343356 TI - Intercepting migraine: results of early therapy with nonspecific and migraine specific agents. AB - Migraine is a pervasive neurologic disorder characterized by recurrent attacks of disabling headache. Despite significant morbidity with impact that may be physical, emotional, social, and economic, treatment of these attacks is often delayed. Patients frequently delay therapy until the more severe or "textbook" symptoms arise, often mistaking the earliest stages as representative of "tension" or "sinus" headaches. Clinicians may recommend deferral of treatment until the more severe levels of pain are seen, perhaps in a misguided attempt to conserve pharmaceutical resources. Patients and clinicians seem more comfortable with perspectives of "being sure it's a migraine" and "not wasting the medication on milder headaches." Therefore, patients and clinicians must learn the latest lessons in migraine: 1) mild migraine usually progresses to more severe levels if left untreated, 2) early treatment is more effective than delayed treatment, 3) early treatment may result in lower rates of adverse events and headache recurrence, and 4) early treatment is cost effective. As clinicians advocate the early treatment of migraine in its mild phase, evidence to support this recommendation has finally become available. I educate my migraineurs to consider each typical headache to be a version of migraine. Most patients with migraine will experience "little" headaches that they often mislabel as tension, sinus, regular, stress, or normal headaches. Instead of these terms, I have them consider their attacks as "small migraines" and "big migraines," with the smaller headaches often evolving into the bigger episodes. Given such a foundation, I advise them to treat at the beginning of the headache, perhaps earlier than they would have previously identified it as a migraine. They must capture the attack while it "whispers migraine" instead of delaying until the attack "shouts migraine." Early treatment of migraine is successful for most patients. However, there are situations in which treatment of the mild phase is not advisable or possible. In patients with frequent or daily migraine, treatment must be reserved for the most disabling attacks. We must advise treatment as soon as the migraine becomes moderate to severe. Certain patients, or certain headaches in some patients, may not progress through a mild phase, perhaps because of rapid escalation or because migraine is already severe upon awakening. Here we encourage migraineurs to treat as soon as possible, often with parenteral formulations of medication. The reaction of the patient (speed of dosing) and the action of the medication (speed of onset of the drug) will ultimately play roles in the successful interception of each attack. PMID- 16343357 TI - Rebound-withdrawal headache (medication overuse headache). AB - Rebound-withdrawal headache (medication overuse headache) is a problem that affects 1% of the population or perhaps more. Work from many countries has shown this to be a worldwide problem. It appears that the rebound-withdrawal headache often occurs on the background of pre-existing migraine or tension headache and may assume the phenotype of chronic migraine or chronic tension-type headache. The key feature of this entity is that the patient initially uses a symptomatic treatment for headache with good result. Use of this medication may increase over time, but as the use increases to more than 10 to 15 days per month, headache frequency may also increase, reaching a point where metabolic processing of the medication and removal from the system triggers another headache. The patient then finds that as use of the medication increases, the frequency and intensity of the headaches also increases, and the overall headache worsens. Once this pattern is established, therapy requires removal of the medication in order for the rebounding process to be terminated. Preventative antimigraine medications will not work in the presence of the rebound phenomenon. Once the rebound withdrawal headache is established, it is difficult to treat. After withdrawal of the medication, there is often a high relapse rate of using the medication again with redevelopment of the rebound-withdrawal headache. Even in patients who do not relapse to drug use, headache usually remains a major problem. Recognition and treatment of rebound headache are key factors in management of these patients. Perhaps more important in regard to this entity is recognizing its existence and preventing patients from entering a pattern of rebound withdrawal, which can be achieved by early attention to their use of analgesic narcotic or migraine-abortive medications. Overall, it is likely that this will continue to be an increasing worldwide problem in managing patients with headache. PMID- 16343358 TI - Headache related to brain tumors. AB - Headache is one of the most common somatic complaints of patients seeking medical care. Most headaches are not of serious cause and can be diagnosed easily with a good history and physical examination. The challenges to the physician are to determine when underlying intracranial pathology may be causing the symptoms and signs, and to identify the few patients in whom a tumor is the cause of the headache. The subject of headache in patients with brain tumors has been reviewed in neurologic textbooks and in several investigations before, as well as after, modern imaging diagnostic techniques became available. Headache can also manifest as an acute or chronic complication of radiation treatment and/or chemotherapy in patients with intracranial neoplasm, but there are few data in the literature specifically addressing this subject. This article provides an overview of headache in patients with primary and secondary brain tumor, including headache characteristics, the putative mechanism for these headaches, the role of diagnostic testing, and the general principles of management. PMID- 16343359 TI - Are the headache guidelines useful in therapeutic decisions for primary care physicians, general neurologists, and headache specialists? AB - The International Headache Society Classification of Headache Disorders has been widely accepted as the gold standard for classification of headache. Initially a research tool, this classification is now increasingly used in the daily practice of headache medicine. Accurate diagnosis is a prerequisite to planning a therapeutic approach. The three commentaries here discuss the use of this tool in the setting of primary care, general neurology, and subspecialty headache medicine. As the Section Editor, I hope these perspectives are helpful to the reader. PMID- 16343360 TI - Diagnosis and management of functional visual deficits. AB - Patients with functional or nonorganic visual disturbances are often seen in general ophthalmology practice and then referred to neurology and/or neuro ophthalmology. The diagnosis of functional vision loss requires comprehensive examinations and diagnostic testing to eliminate possible organic entities. Inconsistencies between examinations may be key to the functional/nonorganic nature. These cases represent some of the most time-consuming diagnoses that an ophthalmologist sees. These patients may be malingering or may have already seen multiple physicians. The more sophisticated patients may go online to learn about an entity that they are trying to mimic. Large amounts of time and money may be spent on diagnostic workups that are thought necessary to prove that there is nothing organic involved. Occasionally there are legal elements of the vision loss, such as minor trauma, in which case the history is key; the complaints tend to resolve when the legal issues are resolved. Another issue to consider is whether someone claiming to be blind is trying to qualify for workmen's compensation or disability. PMID- 16343362 TI - Treatment of chronic facial pain. AB - Facial pain occurs because of damage to the fifth cranial nerve anywhere along its course from its terminal subcutaneous craniofacial branches to the brainstem. Although topical agents may be effective in relieving pain caused by subcutaneous branch damage, systemic oral agents are usually needed to alter or correct deeper trigeminal nociceptive pain caused by damage to the trigeminal nerve further along its course. Antidepressive agents with anti-nociceptive properties, anticonvulsants, and anti-inflammatory agents are most commonly used. Newer agents are beginning to replace the commonly used first-line medications. Combination therapy is popular because it maximizes the effect of each drug while reducing the side effects seen in higher-dose monotherapy. Treatment of secondary clinical depression is very important in the management of patients with facial pain, explaining the beneficial dual role of antidepressants in this condition. Alternative and holistic approaches are also popular, but most are not confirmed by controlled studies at the present time. PMID- 16343361 TI - Treatment of neuro-ophthalmologic manifestations of tuberculosis. AB - In the absence of obvious pulmonary or disseminated tuberculosis, ocular and central nervous system (CNS) tuberculosis may represent a significant diagnostic challenge. Refinements in polymerase chain reaction techniques and neuroimaging have strengthened the battery of tests used to diagnose CNS and ocular tuberculosis, yet in many cases, the diagnosis remains one of exclusion; it may ultimately be determined through exacerbation by anti-inflammatory therapy with subsequent improvement by antitubercular medication treatment. Because of emerging drug resistance, at least a two-drug regimen is required for therapeutic testing and treatment of isolated ocular tuberculosis. If pulmonary or miliary disease coexists, a 6-month, four-drug regimen with isoniazid, rifampin, pyrazinamide, and ethambutol is required for treatment. Tubercular meningitis is treated with the same four-drug regimen for at least 9 to 12 months. Burden of therapeutic compliance rests on the treating physician and public health sector. Best compliance is realized with directly observed therapy. PMID- 16343363 TI - Visual aspects of Behcet's disease. AB - Behcet's disease is a systemic vasculitis characterized by recurrent oral and genital ulcers and ocular inflammation. The disease may also involve the joints, skin, central nervous system, the lungs, and gastrointestinal tract. Ocular manifestations include anterior uveitis, posterior uveitis, macular edema, retinal vasculitis, conjunctival ulcers, optic neuropathy, papilledema, secondary glaucoma, cataract, and neovascularization of iris or retina. The cause of Behcet's disease is unknown, but autoimmune and genetic factors are considered important in its pathogenesis. Promising new imaging techniques and treatments are available to provide better management for patients. This review focuses on recent data on visual aspects, pathogenesis, and the treatment of Behcet's disease. PMID- 16343364 TI - Advances in molecular diagnostics and therapeutics in head and neck cancer. AB - Extensive treatment-related morbidities and stagnant survival rates over the past few decades for patients with squamous cell cancer of the head and neck (SCCHN) emphasize the need for novel diagnostics and therapeutics based on the molecular characteristics of the tumor. The development of an early detection test remains largely preliminary. Much attention has recently been given to saliva-based early detection assays that use accepted tumor markers such as p53 and DNA methylation. Most of these studies have focused on feasibility as opposed to prospective clinical trials. To date, early detection saliva assays have failed to yield a high enough sensitivity and specificity for broad population-based screening. The use of saliva as a noninvasive, inexpensive, and accessible diagnostic substrate remains desirable. Unlike SCCHN diagnostics, molecular-targeted therapies for SCCHN will soon be a reality, with many more compounds in the pipeline. The most promising of these drugs target the epidermal growth factor receptor (EGFR), which is known to be overexpressed in squamous cell carcinomas. Cetuximab, a monoclonal EGFR antibody, has shown efficacy in combination with radiotherapy in advanced SCCHN in a recent phase III trial and is currently being petitioned for US Food and Drug Administration approval. Likewise, erlotinib, an EGFR tyrosine kinase inhibitor, has shown favorable results in phase II trials as monotherapy and in combination with chemotherapy. Gefitinib, another EGFR tyrosine kinase inhibitor, has shown efficacy as monotherapy, in combination with chemotherapy, and with chemoradiotherapy. At least two phase III trials of gefitinib in patients with advanced SCCHN are ongoing. Such low-toxicity, tumor-specific targeting strategies will soon be available for patients with head and neck cancer. The challenge is to establish assays to determine which patients are most likely to benefit from these agents. PMID- 16343365 TI - Pediatric rhabdomyosarcoma of the head and neck. AB - Pediatric rhabdomyosarcoma is not exclusive to the head and neck. However, the unique anatomy of the head and neck requires special consideration and treatment modifications. The low incidence of these tumors has prevented the development of rigorous treatment protocols. Treatment strategies must be individualized on the basis of histopathologic subtype, prognostic indicators, tumor location, tumor extent, available clinical trial data, and hospital resources. The primary treatment of these tumors typically involves a combination of surgery, radiation, and chemotherapy. Advancements in surgical and radiotherapy techniques have reduced patient morbidity, whereas new chemotherapeutic protocols have improved local disease control and overall survival. Because of the infrequency and complexity of these tumors, patients may benefit from referral to centers with a comprehensive multidisciplinary team that has experience treating these tumors in the pediatric population. If possible, patients should be enrolled and treated on the current Children's Oncology Group protocol. PMID- 16343366 TI - Imaging in head and neck cancer. AB - The goals of imaging in head and neck cancer are to establish tumor extent and size, to assess nodal disease, to evaluate for perineural tumor spread, and to distinguish recurrent tumor from post-treatment changes. MRI is the preferred modality for assessment of nasopharyngeal, sinonasal, and parotid tumors, because of better contrast resolution, high frequency of perineural spread, and less prominent motion artifacts. MRI is the best modality to delineate the extent of intraorbital and intracranial extension of malignant tumors. Tumors of the oropharynx, larynx, and hypopharynx are frequently primarily imaged with CT, which is less affected by breathing and swallowing artifacts. MRI is also the initial study of choice for tumors confined to the oral tongue, and possibly also for other oral cavity locations because MRI is superior in detection of tumor spread into the bone marrow. There is no clear advantage of CT or MRI for evaluation of nodal disease. Positron emission tomography (PET) is very sensitive for metastatic lymph nodes that are at least 8 mm in size and is the technique of choice in dubious cases. Imaging-guided biopsies are performed whenever needed. For imaging of treated head and neck cancer, PET scans have been found to generally offer higher sensitivity than MRI or CT. Combined PET/CT may be the modality of choice because it almost completely eliminates the false-positive and false-negative PET findings. Patients with head and neck cancer who are referred to tertiary care centers commonly arrive with cross-sectional images obtained at other institutions. Reinterpretation of these studies by dedicated radiologists frequently leads to changes in findings, which alter treatment and affect prognosis. PMID- 16343367 TI - Second- and third-line treatments in non-small cell lung cancer. AB - Slow but steady progress has been made in the treatment of advanced non-small cell lung cancer. For first-line therapy, multiple chemotherapy combination therapies can extend survival and improve quality of life. And recently, for the first time ever, a noncytotoxic agent, the antivascular endothelial growth factor antibody bevacizumab, has been shown to improve survival when added to chemotherapy. Striking improvements have also been made in second-line treatment. In August 2004, only one agent was US Food and Drug Administration (FDA) approved in this setting, docetaxel, but by the beginning of 2005, two more were available, pemetrexed and erlotinib. All three of these drugs can significantly benefit patients, with 1-year survival in excess of 30%. Choosing between the three agents can be challenging, and this review focuses on the toxicity differences and predictors of response that can help guide this decision. Docetaxel and pemetrexed, both traditional intravenous cytotoxins, are excellent options for patients who have shown some response to first-line chemotherapy, but at this time, no other means exist to determine likelihood of response. When choosing between the two, pemetrexed causes significantly less neutropenia than does docetaxel, at least on the standard every-3-week regimen. With erlotinib, an oral epidermal growth factor receptor (EGFR) inhibitor, there are factors that can predict for response, including little or no smoking history, and adenocarcinoma histology. Therefore, patients who fit these characteristics are good candidates for second-line erlotinib. However, the relationship between response to erlotinib and improved survival remains unclear, and several laboratory analyses that may help further, such as evaluation of EGFR gene copy number, are still under development. Although erlotinib is the only FDA-approved option currently available for third-line therapy, many patients with good performance status may benefit from third-line therapy and beyond. In addition to the approved second-line options, other single-agent chemotherapies to consider for treatment beyond second-line are gemcitabine, irinotecan, and oral topotecan. Many new drugs, including bevacizumab, ZD6474 (AstraZeneca, Wilmington, DE), sorafenib, cetuximab, paclitaxel poliglumex, epothilones, and others, alone or in combination with traditional agents, are currently undergoing investigation and hold great promise. PMID- 16343368 TI - Adjuvant chemotherapy for non-small cell lung cancer. AB - Studies of adjuvant chemotherapy for non-small cell lung cancer (NSCLC) did not provide a consistent disease-free survival or overall survival benefit in the 1980s and early 1990s. However, recently reported studies have changed the practice of NSCLC treatment, for which adjuvant chemotherapy is now considered the standard of care. This review outlines the issues that may have limited the detection of beneficial effects of adjuvant chemotherapy in early trials and provides detailed analysis of the results of recently published trials of adjuvant chemotherapy for NSCLC. PMID- 16343369 TI - Management of small cell lung cancer. AB - Small cell lung cancer (SCLC) is an aggressive type of lung cancer characterized by rapid growth and early metastasis. It is chemosensitive and radiosensitive, yet decades of research investigating multimodality treatments have failed to control or cure this disease in most patients. First-line treatment of limited stage disease consists of chemotherapy (often etoposide/cisplatin or etoposide/carboplatin) combined with thoracic radiation therapy (TRT), followed by prophylactic cranial irradiation to decrease brain metastases as a site of disease progression for those who experience complete remission or a very good partial response to multimodality treatment. In a Japanese trial, the combination of irinotecan and cisplatin had initially shown promise in treating patients with extensive-stage SCLC, but a confirmatory trial in the United States did not find a difference in overall survival with irinotecan/cisplatin versus etoposide/cisplatin. Adding a third drug to the etoposide/cisplatin combination, as well as other triplet therapies, has mostly been ineffective in improving outcomes. Variables in chemotherapy administration, including maintenance therapy, alternating non-cross-resistance regimens, and dose intensification, have not been shown to increase survival at large. In terms of radiation therapy, early administration of TRT concurrent with chemotherapy, and hyperfractionation, have been beneficial in treatment of limited-stage disease. In patients who relapse, second-line therapy options consist of reinduction of previous chemotherapy or administration of a single agent. Targeted biological therapies for SCLC are now being investigated, and although a great deal of research remains to be done, these agents and their derivatives may provide the most hope for future treatment of SCLC. PMID- 16343370 TI - Emerging approaches to advanced bronchioloalveolar carcinoma. AB - Bronchioloalveolar carcinoma (BAC) is a subtype of non-small cell lung adenocarcinoma that has distinct epidemiologic, histologic, radiographic, and clinical features. The strict pathologic definition requires an absence of any invasion through the basement membrane into pulmonary parenchyma, but there is a growing consensus based on recent clinical studies that this diagnosis should be considered to be based on the clinical features of diffuse ground-glass opacities with minimal or no extra-thoracic spread and histology demonstrating adenocarcinoma with a lepidic growth pattern characteristic of BAC, even if there is a component of invasive adenocarcinoma. Although unifocal or even potentially oligometastatic disease is appropriately treated with resection, advanced BAC is generally treated with systemic therapy. However, multifocal BAC may be indolent enough to follow asymptomatic patients without any systemic therapy if patients are comfortable with this approach, because the rate of disease progression may be slow enough to warrant no therapy for many months or even years. For patients who have symptoms and/or clear evidence of progression over a short interval, standard chemotherapy is appropriate, but I would consider treatment with the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) erlotinib to be the most appropriate initial therapy. This is based on the well-documented activity of the EGFR TKIs erlotinib and gefitinib, the latter no longer commercially available in advanced BAC. Advanced BAC is now emerging as an area of significant research, and clinical trials are particularly appealing considerations for such patients. PMID- 16343371 TI - Bronchogenic carcinoma in solid organ transplant recipients. AB - Malignancies are increased in some types of solid organ transplant patients receiving immunosuppressive therapy and are a significant contributor to patient morbidity and mortality. There may be a 100-fold increase in the incidence of de novo neoplasia in this population. The risk of lymphoproliferative malignancies is well appreciated. In contrast, the risk of solid tumors with their consequent morbidity and mortality is less well known, probably because of their common occurrence in the general population. Lung cancer is the most common cause of cancer death in the United States; therefore, lung cancer in patients undergoing organ transplantation would be expected to occur frequently on the basis of chance alone. However, the lung cancer risk is approximately 20 to 25 times that of the general population, with an incidence of 0.28% to 4.1% in patients after heart and lung transplant. Risk factors thought to contribute include cigarette smoking, advanced age at transplantation, and chronic immunosuppressive therapy. The role of transplantation (and consequent therapy) in the development of lung cancer in this high-risk population remains unclear. As in the nontransplant population, adequate screening techniques are lacking, making early diagnosis and treatment a challenge. Despite close follow-up and routine imaging with chest radiography and CT, lung cancers continue to be discovered incidentally and at advanced stages. Treatment is similar to that of patients who are nontransplanted with similar stage, histology, and performance status. PMID- 16343372 TI - Motivational interviewing for improving adherence to antiretroviral medications. AB - Many interventions have been developed to address barriers to antiretroviral medication adherence, but few have focused on motivation, a fundamental component of behavior change. Research on other health behavior changes and a few pilot studies investigating motivational interviewing (MI) for adherence to antiretroviral medication suggests that MI may be highly beneficial by helping to motivate patients with HIV to adhere to their medications. Existing research, although limited, suggests that MI combined with other interventions is feasible and efficacious for improving adherence to antiretroviral medications. With continued development and refinement of antiretroviral adherence interventions that incorporate MI, more persons with HIV infection can be expected to choose to make the difficult changes necessary for them to benefit from antiretroviral therapy. PMID- 16343373 TI - Managing HIV/AIDS in correctional settings. AB - Approximately one quarter of people living with HIV/AIDS in the United States pass through the correctional system, resulting in a burden of infection on the correctional health care system that has challenged correctional and public health officials. The HIV epidemic behind bars results from the high prevalence of HIV risk behaviors among those incarcerated: illicit drug use, untreated mental illness, prostitution, homelessness, and poverty. Challenges to HIV care in correctional settings include management of comorbid conditions, remoteness from HIV care sites, organizational constraints, and access to effective therapies. Despite these challenges, prisoners with HIV have derived considerable benefit from HIV detection and treatment. In order to achieve parity in HIV outcomes among vulnerable populations, effective prison-release programs that incorporate effective case management with effective drug treatment and adherence strategies are required to extend the benefit of highly active antiretroviral therapy as prisoners transition back to community settings. PMID- 16343374 TI - Antiretroviral adherence in a resource-poor setting. AB - Adherence is key to successful antiretroviral therapy (ART). Many countries with increased HIV prevalence and the greatest need for ART have limited health resources. Recent assumptions that the adherence required for successful ART will not be achieved in resource-poor settings have led to calls for caution in expanded access programs. New studies from Africa refute this, showing excellent adherence and virologic outcomes. Major factors contributing to adherence or nonadherence are whether the drug is accessed for free or is self-funded, patient preparedness for use of ART, stigmatization related to being HIV-positive, and ease of use of regimen. PMID- 16343375 TI - The population effectiveness of highly active antiretroviral therapy: are good drugs good enough? AB - Despite the success of highly active antiretroviral therapy (HAART) at reducing mortality from HIV infection, there is evidence that it is not reaching the all of the population that needs it, even in the developed world. To be maximally effective at the population level, all persons with HIV infection must be diagnosed; those persons with an indication for HAART must enter care, must receive HAART, and then must adhere to appointments and, finally, HAART itself. There is considerable evidence that significant portions of the population infected with HIV have not completed all of these steps of HIV care even in the developed world. Although new medications to treat HIV are certainly needed, significant improvements in viral, immunologic, and clinical outcomes could be achieved from improving timely diagnosis, access to HAART, and adherence to appointments, as well as adherence to HAART. PMID- 16343376 TI - HIV and stigma: analysis and research program. AB - Despite advances in our understanding of HIV transmission and optimal treatment of people with HIV infection, stigmatizing attitudes are a significant barrier to HIV prevention and treatment. Several studies demonstrate that stigma directed towards people with HIV infection presents an obstacle to getting tested for HIV, obtaining optimal HIV care, and engaging in safe sex practices. We review the literature on individual and societal factors associated with HIV-associated stigma and propose a framework for intervention design. PMID- 16343377 TI - Physician contributions to disparities in HIV/AIDS care: the role of provider perceptions regarding adherence. AB - Racial/ethnic minorities in the United States are disproportionately affected by HIV/AIDS. In addition to having higher rates of HIV/AIDS, minorities with HIV/AIDS have higher mortality rates than others in the United States. Disparities in the care received by minorities living with HIV/AIDS contribute to these higher mortality rates. This article provides a review of HIV/AIDS health care disparities and explores providers' contributions to these disparities. An important source of provider contribution to disparities appears to be differential prescribing based on perceptions and assumptions that minority patients may have lower adherence to highly active antiretroviral therapy. Literature regarding this topic is reviewed and strategies for reducing disparities are suggested. PMID- 16343378 TI - The methamphetamine epidemic: implications for HIV prevention and treatment. AB - Methamphetamine and related amphetamine compounds are among the most commonly used illicit drugs, with over 35 million users worldwide. In the United States, admissions for methamphetamine treatment have increased dramatically over the past 10 years. Methamphetamine use is prevalent among persons with HIV infection and persons at risk for HIV, particularly among men who have sex with men. In addition to being associated with increased sexual risk behavior, methamphetamine causes significant medical morbidity, including neurologic deficits, cardiovascular compromise, dental decay, and skin infections, all of which may be worsened in the presence of HIV/AIDS. Methamphetamine use may also result in decreased medication adherence, particularly during "binging" episodes. Behavioral counseling remains the standard of treatment for methamphetamine dependence, although the effectiveness of most counseling interventions has not been rigorously tested. Pharmacologic and structural interventions may prove valuable additional interventions to reduce methamphetamine use. PMID- 16343379 TI - Measuring adherence to highly active antiretroviral therapy: implications for research and practice. AB - Highly active antiretroviral therapy (HAART) has radically changed the course of HIV disease, producing substantial reductions in both HIV-related morbidity and mortality. However, the complexity of the typical daily HAART regimen is substantial, and high levels of adherence are essential for complete and long term viral suppression and the avoidance of drug resistance. The complexity of HAART has made the assessment of medication adherence of paramount importance. Even though various methods are in use, each measures only a subset of adherence behaviors, and each measure has limited predictive validity. Given the individual and public health concerns associated with adherence to HAART, there is a need for the continued development and validation of measures of medication adherence. PMID- 16343380 TI - Yellow fever vaccine as a vehicle to better travel medicine. PMID- 16343381 TI - Preflight medical clearance of ill and incapacitated passengers: 3-year retrospective study of experience with a European airline. AB - BACKGROUND: Cabin environment in commercial airline flights may pose specific risks for the ill or incapacitated. International regulations provide a framework for the transportation of passengers for whom specific in-flight equipment or services are needed to minimize possible adverse effects and incidents on board. Little has been published on the characteristics of cases requiring preflight medical clearance for commercial airlines. METHODS: Requests to our medical center for clearance of ill or incapacitated passengers for a medium-sized European airline, within a 3-year period, were systematically reviewed and characterized. RESULTS: For the period of 1999 to 2001, 1,387 requests for medical clearance were made (1 for every 10,935 passengers), 32 (2.3%) of which were denied and 25 (1.8%) of which were deemed unnecessary. Over half (53.6%) of the requests were on behalf of male passengers, and the most common age group ranged from 61 to 70 years (13.3%). Musculoskeletal conditions and limb fractures in particular were the conditions that accounted for the greater number of cases (24.2%), followed by neuropsychiatric (17.6%), cardiovascular (14.5%), respiratory (12.6%), and neoplastic (10.0%) conditions. The most frequent reasons for denial of clearance were recent myocardial infarction and insufficient clinical data. A significant proportion of passengers needed a qualified escort (38.9%) or travel companion (36.7%). Wheelchairs for displacement during boarding or transit (30.4%), in-flight oxygen (28.1%), and stretchers (27.3%) were also frequent requirements. A total of 1,119 passengers (80.7%) required a mean of 2.3 extra needs on board. CONCLUSIONS: The results of this study may help one to better know the nature of medical cases actually requesting clearance for flying. The characteristics of the conditions found suggest that preflight medical clearance may have the potential to prevent the occurrence of disruptive in flight medical incidents and complications. PMID- 16343382 TI - Illnesses in travelers returning from the tropics: a prospective study of 622 patients. AB - BACKGROUND: Although between 8 and 19% of travelers consult a doctor after returning from the tropics, the full spectrum of health complaints is unknown. METHODS: We analyzed the relative frequency of travel-associated health complaints in all travelers (immigrants returning from visiting their home countries, expatriates, business travelers, and tourists) consulting our department between November 2002 and May 2003. RESULTS: A total of 622 patients were studied (256 women, 366 men; median age 38 yr; 45.3% tourists, 33.8% immigrants, 14.1% expatriates, 6.7% business travelers). The median duration of travel was 36.9 days. The main destinations were Africa (57.6%) and Asia (26.4%). A total of 637 diseases were diagnosed, distributed as follows: skin diseases (23.4%), gastrointestinal infections (19.1%), respiratory tract infections (11.5%), malaria (8.8%), schistosomiasis (7.2%), viral hepatitis (4.1%), urinary tract infections (3.5%), sexually transmitted infections (3.5%), tuberculosis (2.7%), dengue fever (2.5%), and others (13.8%). Malaria and intestinal tract infections accounted for 21% and 23% of diagnoses in the 257 febrile travelers, respectively. Overall, 230 diagnoses (36.1%) corresponded to imported tropical diseases, the main imported diseases being malaria, schistosomiasis, amebiasis, gastrointestinal disorders caused by intestinal nematodes, and dengue fever. CONCLUSIONS: Tropical diseases are not the leading cause of consultation in travelers returning from the tropics. Immigrants are the travelers most at risk of common tropical diseases, with the noteworthy exceptions of dengue fever and invasive schistosomiasis. PMID- 16343383 TI - Combined typhoid fever and hepatitis A vaccine: comparison of immunogenicity and safety to concomitant monovalent vaccine over 3 years. AB - BACKGROUND: The safety and immunogenicity of Viatim, a combined hepatitis A (HA) and typhoid fever (Vi) vaccine, were compared with the monovalent component vaccines up to and 1 month after a booster dose at 3 years. METHODS: Healthy, adult volunteers were randomized to receive Viatim (group A, n = 179) or separate HA and Vi vaccines (group B, n = 181); subgroups were boosted after 3 years with Viatim (groups C and D, n = 56 and 46, respectively). Local and systemic reactions were recorded for 28 days postvaccination. Seroconversion and seroprotection rates and geometric mean antibody concentrations were measured at 14 and 28 days, 1, 2, and 3 years postvaccination, and 28 days after the booster dose. RESULTS: Local and systemic safety profiles were equivalent between the two groups. Immediate local reactions were infrequent (1 in group A and 2 in group B). Local reactions, consisting mostly of mild or moderate pain, were least frequent with monovalent HA. Antibody concentrations to both antigens were similar in groups A and B, in which HA seroprotection rates (> or = 20 mIU/mL) were respectively, 98.7% and 100% at day 28, and 99.1% and 99.0% after 3 years, achieving 100% after the booster. Vi seroprotection rates (> or = 1 microg/mL) of 85.2% and 84.9% after 28 days fell to 32.1% and 35.6% after 3 years, increasing to 67.3% and 69.8% after the booster dose. CONCLUSIONS: The combined HA/Vi vaccine, Viatim, had equivalent tolerability and safety and was as rapidly immunogenic as its component monovalent vaccines when given concurrently. A booster dose after 3 years significantly increased antibody levels with some evidence of relative hyporesponsiveness of the typhoid response. PMID- 16343384 TI - High-risk travel abroad overtook low-risk travel from 1999 to 2004: characterization and trends in 2,622 Spanish travelers. AB - BACKGROUND: Travel medicine in Spain is provided by a few specialized centers that do not come under the auspices of the main health system. Some kind of reform is required to avoid common summer collapses and postponements of the service. In contrast to other European countries, neither the exact role nor the responsibilities of general practitioners and primary health care in travel medicine are clearly defined. METHODS: An observational study was performed with retrospective data concerning 2,622 travelers from 1999 to 2004. Although the study was performed at a third-level travel medicine center, continuous contact with and support to general practitioners was maintained throughout the period. RESULTS: International travel was a steadily increasing reality between 1999 and 2004 despite well-known tragic events involving world safety. The number of high risk travels (53.4%) also increased and even overtook low-risk ones (46.6%). This trend was explained as the result of an increasing number of journeys to sub Saharan Africa (14.9%) and those made by traveling immigrants (64.1% of those journeys), which represented a significantly higher proportion of high-risk travels compared with those made by autochthonous subjects (52.1%; p < .001). Moreover, traveling immigrants tend to consult more frequently in periods < 15 days prior to travel than do autochthonous travelers (p < .0001). A substantial number of highly vulnerable travelers, such as pregnant women, infants, elderly people, and immunosuppressed subjects, was found (1.8%). Low-risk travelers who could have been advised and vaccinated by general practitioners were 1,139 (43.4%). CONCLUSIONS: Given the increasing number of travelers undertaking high risk travels abroad, any kind of reinforcement of travel medicine provision in Spain should be considered essential. General practitioners could attend to a significant proportion of low-risk travelers. PMID- 16343385 TI - Effect of maximizing a travel medicine clinic's prevention strategies. AB - BACKGROUND: Even among travelers who undergo evaluation in travel medicine clinics, illnesses develop despite the emphasis placed on prevention. It is possible that travel-associated disease rates may be modified by maximizing access to care and augmenting educational methods of disease prevention. Use of alternative preventive measures such as alcohol hand gel sanitizers may also alter illnesses among travelers. METHODS: We assessed medical outcomes in a travel population cared for in the setting of free vaccinations, medications, and travel medicine consultation, in which personal preventive measures were presented in numerous formats by a physician specializing in infectious diseases. An initial demographic questionnaire was administered at the time of travel consultation. A post-travel telephone interview conducted 2 weeks after return from travel evaluated illness while abroad, illness upon return, and adherence to travel recommendations. An assessment was also performed regarding the utility of an alcohol hand gel sanitizer. RESULTS: One hundred fifty-five travelers were evaluated (primarily older, well-educated US-born travelers, on vacation with family or coworkers). Travelers filled their prescriptions 98% of the time; 77% reported adherence to antimalarial chemoprophylaxis. Sixty-four percent of travelers developed illness abroad, and 20% developed illness upon return. The most frequent complaints were diarrhea and upper respiratory illness. Ten percent of travelers altered their itinerary owing to illness. The use of alcohol hand gel sanitizers did not appear to impact the development of diarrhea or respiratory illnesses. CONCLUSION: In this small group of travelers, access to free consultation, vaccinations, and medications along with presentation of personal protective measures in various formats did not seem to influence the development of illnesses among travelers. Although not rigorously analyzed, alcohol hand gel sanitizers did not seem to alter diarrhea or respiratory tract illness rates. These data highlight the need for new or more effective methods to prevent illness among travelers. PMID- 16343386 TI - Trends in antimalarial prescriptions in Australia from 1998 to 2002. AB - BACKGROUND: Previous research in a number of countries has suggested considerable variability in prescribing patterns of antimalarial drugs. The aim of this study was to investigate the trends in prescription of antimalarial drugs recommended for chemoprophylaxis in Australia from 1998 to 2002. METHODS: In 2005 data was extracted from the online Australian Statistics on Medicines reports published by the Pharmaceutical Benefits Advisory Committee, Drug Utilization Sub-committee, on antimalarials used in Australia from 1998 to 2002. RESULTS: Doxycycline probably remains the malaria chemoprophylaxis of choice prescribed for Australians visiting multiple drug-resistant malarious areas. Over the past 10 years, there has been marked drop in the prescription of less useful antifolate drugs such as pyrimethamine combination antimalarial drugs, especially pyrimethamine plus dapsone, which was withdrawn by 1999. There has also been a reduction in the number of prescriptions for chloroquine, mefloquine, and proguanil. The number of prescriptions for atovaquone and proguanil remain small, although they have increased steadily since its introduction in 2000, in the absence of a recommendation in the prevailing Australian guidelines. CONCLUSIONS: The prescription of antimalarials such as proguanil, chloroquine, mefloquine, and the pyrimethamine-containing compounds reduced considerably between 1998 and 2002. This was probably largely influenced by the availability of antimalarials, increasing resistance, the issuing of updated guidelines for malaria chemoprophylaxis, and continuing education. Newer drugs such as atovaquone plus proguanil may displace older antimalarials, particularly in the prevention of Plasmodium falciparum infection. PMID- 16343387 TI - Quinine sulfate inhibits invasion of Salmonella typhimurium and Shigella flexneri: a preliminary study. AB - BACKGROUND: The incidence of typhoid fever and shigellosis parallels that of malaria, so many individuals who are on antimalarial drugs can be found in areas where these diseases are widespread. We investigated the effect of quinine sulfate on the growth and invasion of Salmonella typhimurium and Shigella flexneri M90T to determine whether people on antimalarials can have secondary gain from some protection against typhoid fever and shigellosis. METHODS: The effect of 50 and 100 microM quinine sulfate on the invasive ability of Salmonella typhimurium and Shigella flexneri M90T into human colon adenocarcinoma-2 (Caco-2) cells was studied during the infection period. The invasive efficiency was expressed as the number of viable internalized bacteria by counting the colony forming units. RESULTS: The invasive ability of Salmonella typhimurium and Shigella flexneri M90T was significantly inhibited by 50 and 100 microM quinine sulfate in a dose-dependent manner (for Salmonella typhimurium) when the drug was added to Caco-2 cell monolayers during the infection period. CONCLUSIONS: Since so many people who are on antimalarial drugs visit and inhabit areas that are endemic to typhoid fever and Shigella infection, a study on the influence of these drugs on the disease is long overdue. Our data indicate that quinine sulfate interferes with the invasion and internalization of Salmonella typhimurium and Shigella flexneri M90T into host cells. Further studies on additional strains/serotypes with other newer antimalarials at various concentrations are needed to verify this effect of quinine sulfate and to draw conclusions on its significance in vivo. PMID- 16343388 TI - Chronic leg ulcer caused by Mycobacterium immunogenum. AB - Rare tropical skin diseases are seen more frequently in Western countries because of the increased popularity of visiting tropical regions. A 55-year-old white man developed a painless leg ulcer after traveling in Guatemala and Belize. A mycobacterium was cultured from a biopsy specimen and was identified as Mycobacterium immunogenum by 16S recombinant deoxyribonucleic acid sequence analysis. The leg ulcer healed after 6 months of compression therapy and hydrocolloids; a hypopigmented depressed scar remained. PMID- 16343389 TI - Malaria in a seaport worker in Haifa. AB - A healthy port worker who had traveled to Azerbaijan 2 years previously was diagnosed with vivax malaria. This is most likely a case of seaport malaria, which has been reported only four times previously. The importance of obtaining a proper occupational, as well as travel, history when evaluating febrile patients is emphasized. PMID- 16343390 TI - Acute rheumatic fever: an unusual cause of fever in a returned traveler. AB - Acute rheumatic fever presenting as fever in a returned traveler has not previously been described. We report one such case. PMID- 16343395 TI - Effect of sodium thiosulphate and cis-diamminedichloroplatinum on FADU tumour cells in nude mice. AB - OBJECTIVES: To study the effect of cis-diamminedichloroplatinum (CDDP) on FADU squamous cell carcinoma cells in a nude mouse model and to determine the effect of sodium thiosulphate (STS) on CDDP activity. METHODS: CD1 nude mice were inoculated with FADU tumour cells to both flanks. They were then randomized to four treatment groups: control, CDDP only, STS only, or CDDP and STS. Tumour growth was measured using calipers and charted at 3-day intervals. RESULTS: Tumour volumes were calculated as an ellipsoid and charted against time. CONCLUSIONS: CDDP inhibited FADU tumour cell growth compared with saline controls (p < .005). The addition of STS did not inhibit the CDDP activity when compared with CDDP-alone activity (p = .989). Compared with saline control solution, STS alone also inhibited tumour growth significantly (p < .005). PMID- 16343396 TI - Tisseel and its effects on wound drainage post-thyroidectomy: prospective, randomized, blinded, controlled study. AB - OBJECTIVE: This randomized, blinded, controlled study examines the effects of fibrin sealant (Tisseel, Laboratoire de production Baxter AG, Vienna, Austria) on wound drainage following thyroidectomy. METHODS: Fifty-six consecutive patients were enrolled in the study. Patients were randomized into Tisseel and non-Tisseel treatment groups. Wound drain output was tallied in 8-hour increments by observers blinded to the treatment groups. RESULTS: Fifty-six patients completed the study. Significant decreases in wound drainage were found in the first 8 hours in the Tisseel group. Eight hours postoperatively, wound output in the Tisseel group was reduced by 44% compared with the non-Tisseel group. A significant decrease in the total drainage over the 64-hour time period of 43% was noted between the treatment and control groups. Post-thyroidectomy wound drainage was reduced and trended to earlier drain removal. No significant changes in the length of hospital stay were noted, nor were postoperative complications encountered in either treatment group. CONCLUSIONS: Fibrin sealants offer a unique opportunity to safely decrease post-thyroidectomy wound drainage. This investigation furthers the evidence that fibrin sealants could safely enable the implementation of drain-free thyroidectomies. PMID- 16343397 TI - Treatment outcome of nasal and paranasal sinus carcinoma. AB - PURPOSE: Most authors recommend aggressive management for sinonasal carcinoma treatment. In an attempt to determine the optimal treatment, we assessed the treatment results of our patients with nasal cavity and paranasal sinus carcinoma. MATERIALS AND METHODS: From January 1980 to December 2001, 40 patients with malignant tumours of the nasal cavity and the paranasal sinuses were treated. The median follow-up was 6 years. Thirty-two patients had tumours originating from the maxillary sinus. Thirteen patients had T1-T2 (32.5%) tumours and 27 patients had T3-T4 (67.5%) tumours. The treatment method was surgery plus radiotherapy in 24 patients (60%) and radiotherapy alone in 16 patients (40%). RESULTS: The 5-year overall survival rate was 61%, whereas it was 65% for T1-T2 disease and 56% for T3-T4 disease. The 5-year local control rate was 58%, whereas it was 75% and 50% (p = .219) for T1-T2 and T3-T4 disease, respectively. In multivariate analysis; localization (p = .016), adjuvant radiotherapy (p = .040), local control (p = .05), and gender (p = .013 for female) were statistically significant factors. CONCLUSION: The prognosis for patients with tumours of the sinonasal area is dependent on localization, tumour stage, and treatment modality. Because the most common site of treatment failure is the primary site, efforts to maximize local control should be undertaken. PMID- 16343398 TI - Assessment of cervical lymph node metastasis with different imaging methods in patients with head and neck squamous cell carcinoma. AB - OBJECTIVE: To determine the predictive value of different imaging methods, computed tomography (CT), magnetic resonance imaging (MRI), ultrasonography (US), and single-photon emission tomography (SPECT),-for cervical node metastasis. DESIGN: Prospective clinical trial. SETTING: An academic otolaryngology department. METHODS: Twenty-three consecutive patients with head and neck malignancy were prospectively evaluated for the presence of cervical lymphadenopathy. All patients underwent clinical, CT, MRI, US, and SPECT examinations. Neck dissection was performed for 31 neck sides, and the results of the preoperative evaluation were confirmed by the surgical and histopathologic findings. MAIN OUTCOME MEASURES: The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated for each method and a comparison of the methods was done. RESULTS: The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of CT, MRI, US, and SPECT were 77.7%, 85.7%, 91.3%, 66.6%, and 80.4%; 59.2%, 92.8%, 94.1%, 54.1%, and 70.7%; 81.4%, 64.2%, 81.4%, 64.2%, and 75.6%; 55.5%, 92.8%, 93.7%, 52.0%, and 68.2%, respectively. Both CT and US were found to be superior to clinical examination. There was no statistically significant difference between US and CT. US was found to be superior to MRI and SPECT in detecting cervical node metastasis. CT was also superior to SPECT. CONCLUSION: Our data show that, despite high specificity rates, especially with SPECT, none of the currently available imaging methods are reliable in evaluating the occult regional metastasis because the negative predictive values of all of these methods are rather low. PMID- 16343399 TI - Sudden sensorineural hearing loss: our experience in diagnosis, treatment, and outcome. AB - OBJECTIVES: To describe our experience concerning sudden sensorineural hearing loss (SSNHL) in a large single-institution series of SSNHL patients and to discuss the results. METHOD: This was a retrospective study, and the charts of 156 consecutive inpatients (65 males, mean age 44 years, range 10-74 years; 91 females, mean age 46 years, range 15-75 years) with the diagnosis of SSNHL from 1987 to 2000 were reviewed. One hundred forty-three of 156 patients received multidrug therapy (plasma expanders, antiaggregants, steroids), whereas only 13 SSNHL patients received hyperbaric oxygen therapy. RESULTS: Old age, vascular and metabolic risk factors, and cigarette smoking do not a have a high prevalence in the SSNHL population. An etiologic factor was detected in 23 of 156 (15%) cases (16 cases of acute infection, 4 cases of neurovascular conflicts, 2 cases of cerebellar angiomas, 1 case of cochleovestibular schwannoma). The outcome was not related to the laterality, age, or hearing loss type. On the contrary, a statistically significant association between poor recovery and male sex, both tinnitus and vertigo, and the initial severity of the hearing loss was observed. CONCLUSIONS: Mostly, SSNHL results in idiopathic disease. At present, diagnostic and therapeutic efforts appear to be inadequate to improve the prognosis of SSNHL. Further studies are needed to obtain better knowledge about the etiopathogenesis of SSNHL so that new therapeutic strategies can be considered in the treatment of this challenging ear disease. PMID- 16343400 TI - Mucormycosis in immunocompetent individuals: an increasing trend. AB - OBJECTIVE: Mucormycosis is a fatal infection of the immunocompromised individual. It is unusual to affect healthy individuals. We report eight such cases of infection caused by this emerging fungal pathogen in healthy patients. Of the eight cases, three were infected with Apophysomyces elegans, again an unusual pathogen causing mucormycosis. STUDY DESIGN: Retrospective case review conducted at a tertiary referral center. METHODS: From 1999 to 2003, eight cases of mucormycosis were managed in otherwise healthy patients. Seven of them were treated with surgery. Clinical presentation, imaging studies, mycologic findings, operative findings at surgery, and postoperative results were evaluated. A review of the literature pertaining to mucormycosis infecting otherwise healthy patients and A. elegans infecting otherwise healthy patients in the nose and paranasal sinuses also was done. RESULTS: Of the eight cases, three were infected with A. elegans, with no history of trauma or any invasive procedure. Seven patients underwent surgical treatment. Histopathologic examination showed broad, sparsely aseptate, thin-walled hyphae and angioinvasion with thrombosis. CONCLUSION: Mucormycosis must be considered in the differential diagnosis of any severe acute headache, sinusitis, or orbital cellulites, not only in immunocompromised patients but also in the absence of any underlying disease. Successful treatment requires tissue debridement and injection of amphotericin B. PMID- 16343401 TI - Effect of recombinant human bone morphogenetic protein 7 on bone density during distraction osteogenesis of the rabbit mandible. AB - OBJECTIVES: To examine whether the quality of bone generation in distraction osteogenesis of the rabbit mandible is enhanced by administering recombinant human bone morphogenetic protein 7 (rhBMP-7) to the lengthened site at the end of the distraction phase. STUDY DESIGN: Prospective randomized study of 24 rabbits undergoing distraction osteogenesis of the mandible, followed by injection of rhBMP-7 or a lactate buffer. SETTINGS: McGill University animal care facility. METHODS: Twenty-four white New Zealand rabbits underwent unilateral mandibular osteotomy and application of an Orthofix uniplanar M-100 fixator (Orthofix Inc., Verona, Italy) to the mandible. Mandibular distraction was carried out for 3 weeks. The animals were divided into three groups: group 1 (eight rabbits), control group: distraction, no injection; group 2 (eight rabbits), study group: distaction + injection of 200 microg of rhBMP7-lactate buffer; group 2 (eight rabbits), comparison group: distraction + injection of lactate buffer. All rabbits were sacrificed 7 weeks after surgery. The mandibles underwent radiology and bone densitometry analysis. MAIN OUTCOME MEASURES: Radiology assessment of bone generation within distracted segments and objective analysis of bone mineral density. RESULTS: Radiology confirmed the presence of bone generate within distraction sites. Densitometry showed a difference in bone mineral density among the three groups (p < .05). CONCLUSION: Local injection of a single dose of 200 microg of rhBMP-7 did enhance bone density when compared with the control group, although not in a statistically significant way when analyzed by an x-ray bone densitometer. This study serves as a starting point for future studies in bone tissue engineering by guiding the strategic administration of rhBMPs and the use of new state-of-the-art technologies, such as micro CT scanning, to analyze in a more detailed way, the effects of rhBMP administration. PMID- 16343402 TI - Is suction drainage an effective means of preventing hematoma in thyroid surgery? A meta-analysis. AB - OBJECTIVE: To evaluate the efficacy of suction drainage in preventing postoperative hematoma formation in thyroid surgery. METHODS: We conducted a meta analysis using only randomized controlled trials in which the incidence of post thyroidectomy hematoma was compared directly in patients with and without suction drains (eight studies since 1980; N = 944). The odds ratio (OR) with respective confidence intervals (CIs) using the fixed effects model was reported. We used an OR < 1.0 as being in favour of treatment (ie, the use of suction drains). RESULTS: In our meta-analysis, there was no statistically significant difference between the rates of post-thyroidectomy hematoma whether or not suction drains were used when the results were combined using a fixed effects model (OR 1.04, 95% CI-1.93), with p = .90. In this comparison, a fixed effects model was used rather than a random effects model because there was no statistically significant heterogeneity (chi2 = 6.26, p = .28). CONCLUSIONS: We conclude that the use of suction drains in thyroid surgery to prevent postoperative hematoma is not evidence based. PMID- 16343403 TI - Laryngotracheoplasty: 30-year follow-up to a pioneering case. PMID- 16343404 TI - Dimensions and outcome of synovial sarcoma of the head and neck: case presentation and review of the literature. PMID- 16343405 TI - [Descending motor deficiency: atypical presentation of Guillain Barre syndrome ]. PMID- 16343406 TI - Lateral sinus thrombosis: serious complication of otitis media. PMID- 16343407 TI - Intraductal papilloma arising from Stensen's duct. PMID- 16343408 TI - Follicular dendritic cell sarcoma: case report and review of current management principles. PMID- 16343409 TI - Familial Zenker's diverticulum: case study and historical perspective. PMID- 16343411 TI - Impairment related to blood amphetamine and/or methamphetamine concentrations in suspected drugged drivers. AB - Experimental studies have investigated effects of low oral doses of amphetamine and methamphetamine on psychomotor functions, while less work has been done on effects of high doses taken by abusers in real-life settings. There are indications that intake of high doses may impair traffic related skills, and that abuse of amphetamines may cause hypersomnolence at the end-of-binge. The present study aimed at investigating the concentration-effect relationship between blood amphetamines concentrations and impairment in a population of real-life users. Eight hundred and seventy-eight cases with amphetamine or methamphetamine as the only drugs present in the blood samples were selected from the impaired driver registry at The Norwegian Institute of Public Health. In each case the police physician had concluded on whether the driver was impaired or not. 27% of the drivers were judged as not impaired, while 73% were judged as impaired. There was a positive relationship between blood amphetamines concentrations and impairment. The relationship reached a ceiling at blood amphetamines concentrations of 0.27 0.53 mg/l. Younger drivers were more often judged impaired than older drivers at similar concentrations. Despite the performance enhancing qualities of amphetamines demonstrated in some low dose laboratory experiments; this study revealed a positive relationship between blood amphetamines concentration and traffic related impairment. PMID- 16343412 TI - Further genetic characterization of the two Trypanosoma cruzi Berenice strains (Be-62 and Be-78) isolated from the first human case of Chagas disease (Chagas, 1909). AB - We describe here an extension of a previous genetic characterization of Trypanosoma cruzi strains (Be-62 and Be-78) isolated from the patient Berenice, the first human case of Chagas disease [Chagas, C., 1909. Nova Tripanomiase humana. Estudos sobre morfologia e o ciclo evolutivo do Schizotrypanum cruzi, n. gen., n. sp., agente etiolojico da nova entidade morbida do homem. Mem. Inst. Oswaldo Cruz 1, 159-218]. We wanted to verify the composition of T. cruzi populations originated from these two isolates. In the present work, 22 enzymatic loci (MLEE), nine RAPD primers and 7 microsatellite loci were analyzed. Clones from both strains were also characterized to verify whether these strains are mono or polyclonal. Be-62 and Be-78 strains were different in 3 out of 22 enzymatic systems, in 3 out of 9 RAPD primers tested and in all microsatellite loci investigated. However, our data suggests that both strains are phylogenetically closely related, belonging to genetic group 32 from Tibayrenc and Ayala [Tibayrenc, M., Ayala, F.J., 1988. Isoenzime variability in Trypanosoma cruzi, the agent of Chagas' disease: genetical, taxonomical, and epidemiological significance. Evolution 42, 277-292], equivalent to zymodeme 2 and T. cruzi II major lineage which, in Brazil, comprises parasites from the domestic cycle of the disease. Microsatellite analyses showed differences between the parental strains but suggested that both populations are monoclonal since each strain and their respective clones showed the same amplification products. PMID- 16343413 TI - Structural analysis of ligand binding and catalysis in chorismate lyase. AB - Chorismate lyase (CL) removes the pyruvyl group from chorismate to provide 4 hydroxybenzoate (4HB) for the ubiquinone pathway. We previously reported the crystal structure at 1.4A resolution of the Escherichia coli CL with bound 4HB product, showing that the product is bound in an internal cavity behind two flaps. To provide a more complete basis for understanding CL's unusual ligand binding properties and mechanism of action, we now report four crystal structures of CL mutants and inhibitor complexes, together with binding and activity measurements and molecular dynamics simulations. First, an ultrahigh resolution (1.0A) crystal structure of the CL*product complex reveals details of a substrate sized internal cavity, also behind the flaps, near the product site. Second, a 2.4A structure of CL complexed with the inhibitor vanillate shows the flaps partly opened relative to their product-bound positions. Third, a 2.0A structure of the G90A mutant with bound product reveals the basis for tighter product binding and kinetic effects of this active site mutation. Fourth, the combination of the G90A mutation with the vanillate inhibitor produces a 1.9A structure containing two inhibitor molecules, one in the product site and the other in the adjacent cavity. The two sites are connected by a short tunnel that is partly open at each end, suggesting that CL may operate via a 2-site or tunnel mechanism. PMID- 16343414 TI - 1,6-Diaminohexane contributes to the hexamethylene bisacetamide-induced erythroid differentiation pathway by stimulating Ca2+ release from inositol 1,4,5 trisphosphate-sensitive stores and promoting Ca2+ influx. AB - Hexamethylene bisacetamide (HMBA) stimulates Ca(2+) signals in murine erythroleukemia (MEL) cells serving as an important component of the HMBA-induced pathway that promotes differentiation to the erythroid phenotype. We observed that 1,6-diaminohexane (DAH) triggered a more rapid and robust increase in MEL cell Ca(2+) levels compared to HMBA and the monodeacetylated N-acetyl-1,6 diaminohexane (NADAH), and that polyamine deacetylase inhibition completely abolished the ability of HMBA and NADAH to induce Ca(2+) signals in MEL cells. Our work indicates that DAH mediates Ca(2+) signal propagation via its ability to activate inositol 1,4,5-trisphosphate (IP(3)) receptors, as we observed similar Ca(2+) release characteristics and heparin sensitivity of DAH and IP(3) in permeabilized MEL cells. Finally, we observed that the DAH-induced Ca(2+) release pathway robustly coupled to a Ca(2+) influx pathway that could be distinguished from thapsigargin-induced Ca(2+) influx by its unusual insensitivity to 2 aminoethoxydiphenyl borate. PMID- 16343416 TI - Disulfide between Cys392 and Cys438 of human serum albumin is redox-active, which is responsible for the thioredoxin-supported lipid peroxidase activity. AB - Human serum albumin (HSA) is an abundant protein found in blood plasma and extracellular fluids. Previously, we found that HSA has a distinct thioredoxin (Trx)-dependent lipid peroxidase activity in the presence of palmitoyl-CoA. In this paper, we identified the redox-active disulfide, which can be specifically reduced by Trx, responsible for the Trx-dependent lipid peroxidase activity. The IIB-III fragment of HSA (Pro299-Leu585) sustained the Trx-dependent lipid peroxidase activity. Chemical modification of the Trx-reduced IIB-III with a thiol-specific modification agent resulted in a complete loss of the peroxidase activity. The analysis of tryptic-peptides derived from the inactivated HSA and IIB-III revealed that Cys392 and Cys438, which exist as an intramolecular disulfide bond in HSA, were preferentially modified in both HSA and IIB-III. Taken together, these results suggested that HSA has a capability to reduce lipid hydroperoxide with the use of Trx as an in vivo electron donor, and that the redox-active disulfide between Cys392 and Cys438 acts as a primary site of the catalysis for the Trx-linked lipid peroxidase activity. PMID- 16343415 TI - Expression, purification, and characterization of cysteine-free mouse P glycoprotein. AB - Cysteine-free mouse MDR3 P-glycoprotein (Pgp) was constructed by mutagenesis of the nine natural Cys to Ala. The Cys-free protein was expressed in Pichia pastoris and purified. Yield, purity, ATPase activity, K(m)(MgATP), and stimulation of ATPase by verapamil, were similar to wild-type mouse Ppg. Mouse Cys-free Pgp was superior in yield and stability to Cys-free human MDR1 Pgp. Mutants Y1040A and Y1040C were constructed in mouse Cys-free Pgp background. Both showed extremely low ATPase activity, strongly-impaired vanadate-trapping of ADP, and reduced photolabeling by 8-azido-ATP. The results are consistent with the conclusion that Tyr-1040 is located in the MgATP-binding site in NBD2 and is required for correct binding and/or orientation of bound MgATP substrate in Pgp as previously suggested by X-ray structures of other ABC transporters and by sequencing of photolabeled Pgp. The results also support our previous conclusion that both catalytic sites must be intact for normal function in Pgp. PMID- 16343417 TI - In vitro and in vivo antimicrobial effects of mastic chewing gum against Streptococcus mutans and mutans streptococci. AB - BACKGROUND AND OBJECTIVE: Dental caries is associated with oral pathogens and Streptococcus mutans (S. mutans) is one of the primary cariogenic organisms. Mastic gum, from Pistacia lentiscus, has been shown to have antibacterial properties. The objective of this study was to determine antibacterial activity of mastic chewing gum against S. mutans and mutans streptococci in vitro and in vivo conditions. SETTING: Cukurova University, Dental School, in 2002. MATERIALS AND METHODS: Antimicrobial activity of mastic gum was evaluated using standard S. mutans strain by disc diffusion method in vitro. Cytotoxicity effect of mastic gum on HEp-2 cells was evaluated by conventional haemocytometer using the trypan blue exclusion method. Clinical studies were then performed on 25 periodontally healthy volunteers. The inhibitory effect of chewing mastic gum against mutans streptococci in saliva was compared to a placebo gum. Saliva samples were taken from the subjects immediately before and after chewing the mastic gum and the placebo gum for 15 min. Additional saliva samples were collected every 30 min. The samples were inoculated onto mitis salivarius-bacitracin agar and incubated for 48 h anaerobically at 37 degrees C. The total number of viable bacteria was then counted. RESULTS: Among tested solvents (chloroform, acetone, petrolium ether and ethanol), it was found that the acetone was found to be more convenient than the others to dissolve the mastic gum. In the cytotoxicity assay, concentrations up to 75 mg/ml of the mastic gum were not toxic for the replication of HEp-2 cells. Thus, lower concentrations of mastic gum (20 and 50 mg/ml) were used for the experiments. In vitro experiments, the diameters of growth inhibition zones of mastic gum were in the range 9.0-27.0 mm. In the clinical trials, the mean number of bacteria in samples taken after chewing the mastic gum and placebo gum were following; at minute 15 was 112 x 10(4)+/-268 x 10(3) and 175 x 10(4)+/-417 x 10(3)cfu/ml, for minute 45 was 85 x 10(4)+/-219 x 10(3) and 165 x 10(4)+/-329 x 10(3)cfu/ml, at minute 75 was 65 x 10(4)+/-100 x 10(3) and 160 x 10(4)+/-216 x 10(3)cfu/ml, at minute 105 was 60 x 10(4)+/-127 x 10(3) and 150 x 10(4)+/-138 x 10(3)cfu/ml, and at minute 135 was 55 x 10(4)+/-65 x 10(3) and 145 x 10(4)+/-354 x 10(3)cfu/ml, respectively. Significantly fewer bacteria was found in saliva samples collected after chewing mastic gum compared to those after chewing paraffin (p<0.001). CONCLUSIONS: This preliminary study showed that mastic gum had significant antibacterial activity against S. mutans and mutans streptococci and it may be a useful adjunct in the prevention of caries. PMID- 16343418 TI - The mediating effects of misinterpretation of intrusive thoughts on obsessive compulsive symptoms. AB - Perfectionism and inflated responsibility have both been identified as risk factors for the development and maintenance of obsessive-compulsive (OC) symptoms. The aim of the present study was to test whether the relationships between these two variables and OC symptoms are mediated by the misinterpretation of intrusive thoughts (MIT). Three hundred and three university students completed the Frost Multidimensional Perfectionism Scale, the Maudsley Obsessional Compulsive Inventory, the Responsibility Attitude Scale, and the Responsibility Interpretations Questionnaire. MIT was found to partially mediate the relationship between responsibility attitudes and OC symptoms. MIT also partially mediated the relationship between concern over mistakes and OC symptoms, even after controlling for responsibility attitudes. Both concern over mistakes and responsibility attitudes were significant predictors of MIT and OC symptoms, but responsibility was the stronger predictor when all of the variables were included in the model. Clinical implications for the treatment of OCD are discussed. PMID- 16343419 TI - The effect of arousal on memory for emotionally-relevant information: a study of skydivers. AB - The aim of the present study was to test memory for information under conditions of extreme arousal in an analogue design to mimic hyperarousal during trauma exposure. Prior to their skydive, 70 novice skydivers were administered measures of fear, heart rate, and anxiety sensitivity. Half the participants learnt a list of words that varied in both relevance to their skydive and valence (irrelevant/relevant, negative/neutral) on the plane (skydivers), and remaining participants learnt an identical list of words on the ground (delayed controls). Following the skydive, participants completed free recall and recognition tasks; heart rate, dissociation, and subjective arousal were also measured. Consistent with an attentional narrowing hypothesis, skydivers recalled comparable a comparable number of relevant words as delayed controls, but significantly less irrelevant words. In terms of recognition performance, skydivers displayed better recognition of relevant words but poorer recognition of irrelevant words than delayed controls. Valence of words did not alter the pattern of results. The implications of the findings are discussed in the context of understanding memory processes under conditions of high threat. PMID- 16343420 TI - Anabolic five subunit-type pyruvate:ferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6. AB - The thermophilic, obligately chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, assimilates carbon dioxide via the reductive tricarboxylic acid cycle. A gene cluster, porEDABG, encoding pyruvate:ferredoxin oxidoreductase (POR), which plays a key role in this cycle, was cloned and sequenced. The nucleotide sequence and the gene organization were similar to those of the five subunit-type 2-oxoglutarate:ferredoxin oxidoreductase from this strain, although the anabolic POR had been previously reported to consist of four subunits. A small protein (8 kDa) encoded by porE, which had not been detected in the previous work, was identified in the purified recombinant POR expressed in Escherichia coli, indicating that the enzyme is also a five-subunit type. Incorporation of PorE in the wild-type POR enzyme was confirmed by immunological analysis. PorA, PorB, PorG, and PorE were similar to the alpha, beta, gamma, and delta subunits of the four subunit-type 2-oxoacid oxidoreductases, respectively, and had conserved specific motifs. PorD had no specific motifs but was essential for the expression of the active enzyme. PMID- 16343421 TI - Mdm2-mediated pRB downregulation is involved in carcinogenesis in a p53 independent manner. AB - Mdm2 promotes ubiquitination of the tumor suppressor p53 and can function as an oncogene by largely downregulating p53. Although a p53-independent role of Mdm2 has been reported, the underlying mechanism remains unclear. In the present study, we indicated that Mdm2 is involved in p53-independent carcinogenesis via downregulation of pRB. Expression of pRB showed an apparent inverse correlation with Mdm2 expression in 30 patients with non-small cell lung cancer. There were some cases with the p53 mutations in which a high level of Mdm2 and a low level of pRB were expressed. Mdm2 promoted ubiquitination of pRB in cells without wild type p53. Furthermore, pRB-mediated G1 arrest in a p53-deficient cell line, SRB1, was significantly enhanced by a mutant Mdm2 that lacks pRB ubiquitination activity. Soft-agar colony formation activity of p53-knockout MEF was increased by wild-type Mdm2 but not mutant Mdm2. These findings suggest that overexpression of Mdm2 can perturb a RB pathway regardless of the p53 gene status, promoting carcinogenesis. PMID- 16343422 TI - BCR-crosslinking induces a transcription of protein phosphatase component G5PR that is required for mature B-cell survival. AB - BCR-crosslinking triggers activation-induced cell death (AICD) selectively in the restricted stage of B-cell differentiation. We examined the transcription of a protein phosphatase subunit G5PR in immature and mature B-cells, because absence of this factor augmented cell sensitivity to AICD, associated with increased activation of JNK and Bim. BCR-crosslinking-induced G5pr transcription in AICD resistant mature splenic IgM(lo)IgD(hi) B-cells but not in AICD susceptible immature IgM(hi)IgD(lo) B-cells. Thus, G5pr induction correlated with the prevention of AICD; High in mature splenic CD23(hi) B-cells but low in immature B cells of neonatal mice, sub-lethally irradiated mice, or xid mice. Lack of G5pr upregulation was associated with the prolonged activation of JNK. The G5pr cDNA transfection protected an immature B-cell line WEHI-231 from BCR-mediated AICD. The differential expression of G5PR might be responsible for the antigen dependent selection of B-cells. PMID- 16343423 TI - Prion protein induced signaling cascades in monocytes. AB - Prion proteins play a central role in transmission and pathogenesis of transmissible spongiform encephalopathies. The cellular prion protein (PrP(C)), whose physiological function remains elusive, is anchored to the surface of a variety of cell types including neurons and cells of the lymphoreticular system. In this study, we investigated the response of a mouse monocyte/macrophage cell line to exposure with PrP(C) fusion proteins synthesized with a human Fc-tag. PrP(C) fusion proteins showed an attachment to the surface of monocyte/macrophages in nanomolar concentrations. This was accompanied by an increase of cellular tyrosine phosphorylation as a result of activated signaling pathways. Detailed investigations exhibited activation of downstream pathways through a stimulation with PrP fusion proteins, which include phosphorylation of ERK(1,2) and Akt kinase. Macrophages opsonize and present antigenic structures, contact lymphocytes, and deliver cytokines. The findings reported here may become the basis of understanding the molecular function of PrP(C) in monocytes and macrophages. PMID- 16343424 TI - Immune suppressor factor confers stromal cell line with enhanced supporting activity for hematopoietic stem cells. AB - Immune suppressor factor (ISF) is a subunit of the vacuolar ATPase proton pump. We earlier identified a short form of ISF (ShIF) as a stroma-derived factor that supports cytokine-independent growth of mutant Ba/F3 cells. Here, we report that ISF/ShIF supports self-renewal and expansion of primary hematopoietic stem cells (HSCs). Co-culture of murine bone marrow cells with a stromal cell line overexpressing ISF or ShIF (MS10/ISF or MS10/ShIF) not only enhanced their colony forming activity and the numbers of long-term culture initiating cells, but also maintained the competitive repopulating activity of HSC. This stem cell supporting activity depended on the proton-transfer function of ISF/ShIF. Gene expression analysis of ISF/ShIF-transfected cell lines revealed down-regulation of secreted frizzled-related protein-1 and tissue inhibitor of metalloproteinase 3, and the restoration of their expressions in MS10/ISF cells partially reversed its enhanced LTC-IC supporting activity to a normal level. These results suggest that ISF/ShIF confers stromal cells with enhanced supporting activities for HSCs by modulating Wnt-activity and the extracellular matrix. PMID- 16343425 TI - Induction of calcium influx from extracellular fluid by beauvericin in human leukemia cells. AB - Beauvericin, a cyclic hexadepsipeptide, is a mycotoxin that can induce cell death in human lymphoblastic leukemia CCRF-CEM cells. Our previous data have shown that beauvericin induces cell death in CCRF-CEM cells in a dose- and time-dependent manner, and that this beauvericin-induced cell death can be prevented by administration of intracellular calcium chelator-BAPTA. Therefore, the intracellular Ca2+ concentration ([Ca2+]i) may play an important role in beauvericin-induced cell death in CCRF-CEM cells. In this study, the effect of beauvericin on [Ca2+]i and the possible mechanism responsible for the changes of [Ca2+]i in CCRF-CEM cells were investigated. Beauvericin caused a rapid and sustained [Ca2+]i rise in a dose-dependent manner. Excess extracellular Ca2+ facilitated beauvericin-induced [Ca2+]i rise by adding 1 mM CaCl2 in the bathing medium. On the other hand, beauvericin-induced [Ca2+]i rise was prevented in Ca2+ free Tyrode's solution by 200 microM EGTA. In addition, beauvericin-induced [Ca2+]i rise was also attenuated by intracellular Ca2+ chelator-BAPTA/AM. It is worthy to note that neither the voltage-dependent Ca2+ channel blocker, nimodipine, nor depletion of intracellular Ca2+ with thapsigargin, an endoplasmic reticulum Ca2+ pump inhibitor, has any effect on beauvericin-induced [Ca2+]i rise. The data from present study indicate that beauvericin acts as a potent Ca2+ mobilizer by stimulating extracellular Ca2+ influx CCRF-CEM cells. PMID- 16343426 TI - Ras regulates neuronal polarity via the PI3-kinase/Akt/GSK-3beta/CRMP-2 pathway. AB - The establishment of a polarized morphology is an essential event in the differentiation of neurons into a single axon and dendrites. We previously showed that glycogen synthase kinase-3beta (GSK-3beta) is critical for specifying axon/dendrite fate by the regulation of the phosphorylation of collapsin response mediator protein-2 (CRMP-2). Here, we found that the overexpression of the small GTPase Ras induced the formation of multiple axons in cultured hippocampal neurons, whereas the ectopic expression of the dominant negative form of Ras inhibited the formation of axons. Inhibition of phosphatidylinositol-3-kinase (PI3-kinase) or extracellular signal-related kinase (ERK) kinase (MEK) suppressed the Ras-induced formation of multiple axons. The expression of the constitutively active form of PI3-kinase or Akt (also called protein kinase B) induced the formation of multiple axons. The overexpression of Ras prevented the phosphorylation of CRMP-2 by GSK-3beta. Taken together, these results suggest that Ras plays critical roles in establishing neuronal polarity upstream of the PI3-kinase/Akt/GSK-3beta/CRMP-2 pathway and mitogen-activated protein kinase cascade. PMID- 16343427 TI - Identification of a novel male germ cell-specific gene TESF-1 in mice. AB - Mammalian spermatogenesis is precisely regulated by many germ cell-specific factors. In search for such a germ cell-specific factor, we have identified a novel mouse gene testis-specific factor 1 (TESF-1). Messenger RNA of TESF-1 was found only in the testis and its expression appeared to be regulated in a developmental manner. Further analysis demonstrated that the expression of TESF-1 was specifically in male germ cells, supported by the observation that we were not able to detect the TESF-1 mRNA from at/at homozygous mutant testes, which lack germ cells. The deduced amino acid sequence of TESF-1 contains a leucine zipper motif, a potential nuclear localization signal, and two cAMP- and cGMP dependent protein kinase phosphorylation sites. The green fluorescent protein (GFP)-tagged TESF-1 fusion protein was expressed in COS-7 cells and localized primarily in the nucleus. Taken together, these results indicate that TESF-1 is a novel male germ cell-specific gene, and its protein product may function as a nuclear factor involved in the regulation of spermatogenesis. PMID- 16343428 TI - The refractory periods and threshold potentials of sequential spikes measured by whole-cell recording. AB - Neurons in the central nervous system are thought to program neural language via firing sequential spikes for guiding animal behaviors. The quantitative profiles of spike intrinsic properties are critically important to understand spike programming. We developed approaches with whole-cell recordings to measure the threshold potentials and refractory periods (RPs) of sequential spikes, and to analyze the relationships of these factors with spike timing precision and capacity at the regular-spiking and fast-spiking neurons in cortical slice. The RPs and threshold potentials of sequential spikes at these two groups of neurons are different and are linearly correlated with spike timing precision and capacity. These data suggest that RPs and threshold potentials essentially navigate the spike programming for the precise and loyal encoding of meaningful neural signals. Our study provides the avenues for decoding the spectrum of the neural signals quantitatively. PMID- 16343429 TI - Microtubule-binding properties of dynactin p150 expedient for dynein motility. AB - Dynactin is a hetero-oligomeric protein complex that has an important role in dynein-based intracellular transport. The expressed N-terminal fragments of dynactin p150 bound to microtubules in the ratio of one to one tubulin dimer, independent from the binding of dynein stalk head. Single molecule observation revealed that these fragments moved around on microtubules by Brownian motion. When the dynein-dynactin complex moves on microtubules, p150 can support dynein to maintain contact with microtubules and does not interfere with the motility of dynein, and thus, the dynein-dynactin complex can efficiently achieve long distance carriage of the cargo. PMID- 16343430 TI - Analysis of gene expression in single human oocytes and preimplantation embryos. AB - Little is known about the gene expression in human oocytes and early embryo development because of the rare availability of the materials. The recent advancement of biotechnology has allowed one to analyze the gene expression in single human oocytes and preimplantation embryos. Gene expression of human lactate dehydrogenase isozymes (LDH-A, LDH-B, and LDH-C) and small ubiquitin-like modifier isoforms (SUMO-1, SUMO-2, and SUMO-3) in four oocytes, two 4-cell and three 8-cell embryos was studied using the reverse transcription-polymerase chain reaction. The mRNAs for SUMO-1, SUMO-2, SUMO-3, and LDH-B (heart) were detected in all of oocytes, 4- and 8-cell embryos. The mRNA for LDH-A (muscle) was detected in two of four oocytes and one of three 8-cell embryos. However, the mRNA for testis-specific LDH-C was not detected at all as expected. A cDNA microarray containing 9600 cDNA spots was used to investigate differential expression of human genes in oocyte, 4-cell and 8-cell embryos. The expression of 184, 29, and 65 genes was found to have a value more than twofold above the median value of all genes expressed in oocyte, 4- and 8-cell embryos, respectively, indicating that the expression of some zygotic genes had already occurred at 4-cell embryo. PMID- 16343431 TI - Luteolin, a flavonoid, inhibits AP-1 activation by basophils. AB - Flavonoids including luteolin, apigenin, and fisetin are inhibitors of IL-4 synthesis and CD40 ligand expression by basophils. This study was done to search for compounds with greater inhibitory activity of IL-4 expression and to clarify the molecular mechanisms through which flavonoids inhibit their expression. Of the 37 flavonoids and related compounds examined, ayanin, luteolin, and apigenin were the strongest inhibitors of IL-4 production by purified basophils in response to anti-IgE antibody plus IL-3. Luteolin did not suppress Syk or Lyn phosphorylation in basophils, nor did suppress p54/46 SAPK/JNK, p38 MAPK, and p44/42 MAPK activation by a basophilic cell line, KU812 cells, stimulated with A23187 and PMA. However, luteolin did inhibit phosphorylation of c-Jun and DNA binding activity of AP-1 in nuclear lysates from stimulated KU812 cells. These results provide a fundamental structure of flavonoids for IL-4 inhibition and demonstrate a novel action of flavonoids that suppresses the activation of AP-1. PMID- 16343432 TI - Interaction of glucose transporter 1 with anion exchanger 1 in vitro. AB - The facilitative glucose transporter 1 (GLUT1) mediates the passive diffusion of d-glucose across the cell membrane, providing the energy resource in glycolysis in the erythrocytes. Anion exchanger 1 (band 3) is another important membrane protein that mediates rapid exchange of CO(2) through Cl(-)/HCO(3)(-) exchange across the erythrocyte membrane. For verifying the presumption over a decade that GLUT1 and band 3 in the erythrocyte would be interacting with each other, we cloned and expressed both the cytoplasmic domains of GLUT1 and band 3 in Escherichia coli, and tested their binding ability. By coimmunoprecipitation we found that among the tested N-terminal, C-terminal, and loop fraction of GLUT1, only the C-terminal of GLUT1 can interact with cytoplasmic domain of band 3. The interaction was further verified by coimmunoprecipitation and pull-down assay using both proteins as bait and target. These results showed that GLUT1 and band 3 form a protein complex that can regulate the activities of the proteins within it. PMID- 16343433 TI - In vitro and in vivo characterization of a novel CCR3 antagonist, YM-344031. AB - Eosinophils play a prominent proinflammatory role in a broad range of diseases, including atopic dermatitis and asthma. Eotaxin-1 and its receptor CCR3 are implicated in the recruitment of eosinophils from blood into inflammatory tissues, therefore inhibition of Eotaxin-1/CCR3 interaction may have therapeutic potential for allergic inflammation with eosinophil infiltration. YM-344031, a novel and selective small molecule CCR3 antagonist, potently inhibited ligand binding (IC(50)=3.0nM), ligand-induced Ca(2+) flux (IC(50)=5.4nM), and the chemotaxis of human CCR3-expressing cells (IC(50)=19.9nM). YM-344031 (1-10mg/kg) orally administered to cynomolgus monkeys significantly inhibited Eotaxin-1 induced eosinophil shape change in whole blood. Additionally, orally administered YM-344031 (100mg/kg) prevented both immediate- and late-phase allergic skin reactions in a mouse allergy model. YM-344031 therefore has potential as a novel and orally available compound for the treatment of allergic inflammation, such as atopic dermatitis and asthma. PMID- 16343434 TI - Biochemical properties of the Vibrio harveyi CgtAV GTPase. AB - Bacteria encode a number of relatively poorly characterized GTPases, including the essential, ribosome-associated Obg/CgtA proteins. In contrast to Ras-like proteins, it appears that the Obg/CgtA proteins bind guanine nucleotides with modest affinity and hydrolyze GTP relatively slowly. We show here that the Vibrio harveyi CgtA(V) exchanges guanine nucleotides rapidly and has a modest affinity for nucleotides, suggesting that these features are a universal property of the Obg/CgtA family. Interestingly, CgtA(V) possesses a significantly more rapid GTP hydrolysis rate than is typical of other family members, perhaps reflecting the diversity and specificity of bacterial ecological niches. PMID- 16343435 TI - PKCdelta modulates p21WAF1/CIP1 ability to bind to Cdk2 during TNFalpha-induced apoptosis. AB - Cyclin-dependent kinase 2 (Cdk2) activity is thought to be involved in cell death associated chromatin condensation and other manifestations of apoptotic death. Here we show that during TNFalpha-induced apoptosis, PKCdelta is activated in a caspase-3-dependent manner and phosphorylates p21(WAF1/CIP1), a specific cyclin dependent kinase inhibitor, on (146)Ser. This residue is located near a cyclin binding motif (Cy2) that plays an important role in the interaction between p21(WAF1/CIP1) and Cdk2, and its phosphorylation modulates the ability of p21(WAF1/CIP1) to associate with Cdk2. The phosphorylation of p21(WAF1/CIP1) is temporally related to the activation kinetics of Cdk2 activity during the apoptosis. We propose that during TNFalpha-induced apoptosis, PKCdelta-mediated phosphorylation of p21(WAF1/CIP1) at (146)Ser attenuates the Cdk2 binding of p21(WAF1/CIP1) and thereby upregulates Cdk2 activity. PMID- 16343437 TI - Splicing variant of Cdc42 interacting protein-4 disrupts beta-catenin-mediated cell-cell adhesion: expression and function in renal cell carcinoma. AB - We have identified an alternative splicing variant in the Cdc42-interacting protein 4 (CIP4) gene in patients with renal cell carcinoma (RCC); almost 50% of the RCCs examined showed an aberrant splicing event in reverse transcription-PCR and the insertion of 19 nucleotides derived from intron9 based on a sequence analysis. This variant (CIP4-V) encodes a premature stop codon, resulting in the loss of a tyrosine phosphorylation site, the Cdc42 binding domain, and the SH3 domain. In this report, we show that overexpression of CIP4-V causes the formation of ubiquitinated aggresomes and a loss of cell-cell adhesion. We determined that CIP4-V increased the beta-catenin tyrosine phosphorylation levels that mediate Fer/Fyn tyrosine kinases and induced beta-catenin mistrafficking from cell membrane to cytoplasmic aggresome. These results indicate that CIP4 is critical for beta-catenin-mediated cell-cell adhesion and may be an important aspect of its functional contribution to RCC, especially with regard to metastasis and invasiveness. PMID- 16343436 TI - A novel response element confers p63- and p73-specific activation of the WNT4 promoter. AB - The p53 tumor suppressor gene family consists of three genes, p53, p63, and p73. p53 family proteins share high homology in their DNA-binding domains but exhibit diverse biological functions. In this study, we demonstrated differential target gene activation by specific p53, p63, and p73 induction in Saos2 cells by oligonucleotide microarray expression analysis. We further analyzed the WNT4 promoter, which was induced by p63 and p73 but not p53, in order to clarify the mechanism of differential target gene activation between the three family members. Luciferase analysis showed that the WNT4 promoter harbors two p63/p73 response elements, designated RE1 and RE2. RE1 resembles the canonical p53 response element (tandem repeats of RRRCWWGYYY), located between -141 and -121, while RE2 consists of a GC-rich sequence further downstream. Neither response element alone was able to confer transcriptional activity. It is thus likely that both RE1 and RE2 are necessary in rendering p63/p73-specific activation of the WNT4 promoter. PMID- 16343438 TI - FHL2 mediates p53-induced transcriptional activation through a direct association with HIPK2. AB - To understand the molecular mechanism underlying HIPK2 regulation of the transcriptional activation by p53, we sought to identify the protein that interacts with HIPK2. From our yeast two-hybrid screen, we found that four and a half LIM domains 2 (FHL2) could bind to the C-terminal half of HIPK2. Further assays in yeast mapped the minimal interaction domain to amino acids 812-907 in HIPK2. The interaction was confirmed using a GST pull-down assay in vitro, and an immunoprecipitation (IP) assay and fluorescence microscopy in vivo. FHL2 alone spread throughout both the cytoplasm and nucleus but was redistributed to dot like structures in the nucleus when HIPK2 was coexpressed in HEK293 cells. When tethered to the Gal4-responsive promoter through the Gal4 DBD fusion, FHL2 showed autonomous transcriptional activity that was enhanced by wild-type HIPK2, but not by the kinase-defective mutant. In addition, FHL2 increased the p53-dependent transcriptional activation and had an additive effect on the activation when coexpressed with HIPK2, which was again not observed with the kinase-defective mutant of HIPK2. Finally, we found a ternary complex of p53, HIPK2, and FHL2 using IP, and their recruitment to the p53-responsive p21Waf1 promoter in chromatin IP assays. Overall, our findings indicate that FHL2 can also regulate p53 via a direct association with HIPK2. PMID- 16343439 TI - Activation of connective tissue growth factor gene by the c-Maf and Lc-Maf transcription factors. AB - The Maf family of transcription factors is expressed during development of various organs and tissues, and is involved in a variety of developmental and cellular differentiation processes. We previously found that c-maf and mafB are strongly expressed in hypertrophic chondrocytes during cartilage development. Connective tissue growth factor (CTGF) is also expressed in hypertrophic chondrocytes. Adenovirus mediated introduction of c-maf gene into the mouse fibroblast cell line C3H10T1/2 strongly induced CTGF expression. CTGF can be induced by TGF-beta via the SMAD pathway; however, the c-Maf could not induce TGF beta, nor could TGF-beta induce the c-Maf, suggesting that activation of CTGF by Maf is TGF-beta independent. Reporter transfection analysis using C3H10T1/2 cells shows that c-Maf stimulates a CTGF reporter gene. Lc-Maf, a splice variant of c Maf containing an extra 10 amino acids in the carboxyl terminus, was a stronger inducer of the CTGF reporter gene than c-Maf. Chromatin immunoprecipitation analysis showed that c-Maf binds to the promoter region of the CTGF gene, indicating that Maf directly activates the CTGF gene. Taken together, these data indicate that the CTGF gene is a target of c-Maf and Lc-Maf in cartilage development. PMID- 16343440 TI - Identification of a novel splice variant of X-linked inhibitor of apoptosis associated factor 1. AB - XAF1 (XIAP-associated factor 1) binds to XIAP and blocks its anti-apoptotic activity. It has been reported that XAF1 is mainly expressed in normal tissues but is missing or present at low levels in most cancer cell lines, which implies a tumor-suppressing function. In the present study we describe the identification of a novel splice variant of human XAF1, designated XAF1C, which contains a cryptic exon. Incorporation of this exon (exon 4b) into the mRNA introduces an in frame stop codon, resulting in a shortened open-reading frame (ORF) of 495 nucleotides. This ORF is predicted to encode a 164 amino acid (AA) protein lacking the C-terminal domain of the previously described XAF1(A), but containing a unique 24 AA carboxy terminus. Like XAF1(A), XAF1C mRNA expression was detected in a variety of human cancer cell lines and also in normal human tissues. The ratio of XAF1(A) and XAF1C mRNA expression differs amongst the cell lines tested, suggesting differential mRNA stabilities and/or the existence of tissue- or cell type-specific splicing regulation. In transfected cells, xaf1c encodes a truncated protein of 18kDa, which is distributed primarily in the nucleus. PMID- 16343441 TI - Activation of transcriptional activities of AP-1 and SRE by a new zinc-finger protein ZNF641. AB - Mitogen-activated protein kinases (MAPKs) are evolutionarily conserved enzymes in cell signal transduction connecting cell-surface receptors to critical regulatory targets within cells and control cell survival, adaptation, and proliferation. Previous studies revealed that zinc-finger proteins are involved in the regulation of the MAPK signaling pathways. Here, we report the identification and characterization of a novel human zinc-finger protein, ZNF641. The cDNA of ZNF641 is 4.9kb, encoding 438 amino acids in the nucleus. The protein is highly conserved in evolution across different vertebrate species from mouse to human. Northern blot analysis indicates that ZNF641 is expressed in most of the examined human tissues, with a high level in skeletal muscle. Overexpression of pCMV-Tag2B ZNF641 in the COS-7 cells activates the transcriptional activities of AP-1 and SRE. Deletion analysis indicates that the linker between KRAB box and C(2)H(2) type zinc-fingers represents the basal activation domain. These results suggest that ZNF641 may be a positive regulator in MAPK-mediated signaling pathways that lead to the activation of AP-1 and SRE. PMID- 16343442 TI - Expression of human CMP-N-acetylneuraminic acid synthetase and CMP-sialic acid transporter in tobacco suspension-cultured cell. AB - Plant cells have no beta1,4-galactosylated and sialylated glycan, which plays important roles in biological functions in animal cells. Previously, we generated transgenic tobacco BY2 suspension-cultured cells that produced human beta1,4 galactosyltransferase [N.Q. Palacpac, S. Yoshida, H. Sakai, Y. Kimura, K. Fujiyama, T. Yoshida, T. Seki, Stable expression of human beta1,4 galactosyltransferase in plant cells modifies N-linked glycosylation pattern, Proc. Natl. Acad. Sci. USA 96 (1999) 4692-4697]. In this study, we introduced two critical genes encoding human CMP-N-acetylneuraminic acid synthetase and CMP sialic acid transporter into tobacco suspension-cultured cell to pave a route for sialic biosynthetic pathway. The recombinant human proteins showed their biological activities. These results show that the plant cell can be a useful bioreactor for the production of mammalian glycoproteins. PMID- 16343443 TI - RNA complementary to the 5' UTR of mRNA triggers effective silencing in Saccharomyces cerevisiae. AB - Conditional silencing of target genes in Saccharomyces cerevisiae by antisense RNAs expressed in vivo has been challenged. The MFalpha1::lacZ fusion present in S. cerevisiae SF51-3 was chosen as a model target, and fragments of this gene were cloned in reverse orientation into the expression vector pYES2, bearing the GAL1 promoter. Among the different antisense constructs tested, only the one complementary to the 5' UTR of target mRNA featured effective silencing. Nevertheless, the expression in vivo of this antisense RNA could not be properly tuned by the absence or presence of galactose in the culture medium. Accordingly, conditional silencing could not be attained by this antisense hosted into pYES2. On the contrary, cloning the same antisense construct into the expression vector pSAL4 yielded a fully conditional silencing linked to the control of antisense expression by the absence or presence of Cu(2+) into the culture medium. PMID- 16343444 TI - Characterization of the N-deacetylase domain from the heparan sulfate N deacetylase/N-sulfotransferase 2. AB - Heparin and heparan sulfate are linear sulfated polysaccharides that exert a multitude of biological functions. Heparan sulfate glucosaminyl N-deacetylase/N sulfotransferase isoform 2 (NDST-2), a key enzyme in the biosynthesis of heparin, contains two distinct activities. This bifunctional enzyme removes the acetyl group from N-acetylated glucosamine (N-deacetylase activity) and transfers a sulfuryl group to the unsubstituted amino position (N-sulfotransferase activity). The N-sulfotransferase activity of NDST has been unambiguously localized to the C terminal domain of NDST. Here, we report that the N-terminal domain of NDST-2 retains N-deacetylase activity. The N-terminal domain (A66-P604) of human NDST-2, designated as N-deacetylase (NDase), was cloned as a (His)(6)-fusion protein, and protein expression was carried out in Escherichia coli. Heparosan treated with NDase contains N-unsubstituted glucosamine and is highly susceptible to N sulfation by N-sulfotransferase. Our results conclude that the N-terminal domain of NDST-2 contains functional N-deacetylase activity. This finding helps further elucidate the mechanism of action of heparan sulfate N-deacetylase/N sulfotransferases and the biosynthesis of heparan sulfate in general. PMID- 16343445 TI - Potencies of topical glucocorticoids to mediate genomic and nongenomic effects on human peripheral blood mononuclear cells. AB - Several different genomic and nongenomic mechanisms are known to mediate the important anti-inflammatory and immunomodulatory effects of glucocorticoids (GC). Genomic effects are the most important while the clinical relevance of nongenomic actions is still a matter of debate. We therefore investigated whether beclometasone and clobetasol are particularly suitable for topical application because of their specific spectrum of genomic and nongenomic actions. For these purposes we compared effects on oxygen consumption as measured with a Clark electrode (nonspecific nongenomic glucocorticoid effects), on interleukin-6 synthesis by means of ELISA (genomic effects) and on apoptosis using flow cytometry (nongenomic and genomic effects) in quiescent and mitogen-stimulated PBMC. Beclometasone and clobetasol indeed had stronger effects on the oxygen consumption of quiescent and stimulated cells at lower concentrations (10(-10) and 10(-8) M) but were less potent at higher concentrations (10(-5) and 10(-4) M) in comparison with dexamethasone. Also in terms of genomic potency, topical GC were more effective than dexamethasone at 10(-10) and 10(-8) M but gave similar results at higher concentrations. The ability of all three GC to induce apoptosis was found to be concentration-dependent and similar at concentrations between 10( 8) and 10(-5) M. But, compared with 10(-4) M dexamethasone, topical GC at 10(-4) M were significantly more effective at inducing apoptosis in both PBMC and Jurkat T-cells. These results show that topical GC have different concentration- (genomic/nongenomic) effect--ratios compared with dexamethasone: besides to the well-known genomic effects there are also significant nongenomic effects of topical glucocorticoids that already at low concentrations might be more therapeutically relevant in certain clinical conditions than currently assumed. PMID- 16343446 TI - Anticancer activity of the lanthanum compound [tris(1,10 phenanthroline)lanthanum(III)]trithiocyanate (KP772; FFC24). AB - Aim of this study was to investigate the anticancer properties of the new lanthanum compound [tris(1,10-phenanthroline)lanthanum(III)]trithiocyanate (KP772; FFC24). In vitro, growth inhibition by KP772 was comparable for >60 tumour cell models with IC50 values generally in the low microM range. KP772 induced tumour cell apoptosis indicated by chromatin condensation, caspase substrate cleavage and mitochondrial membrane depolarisation. DNA is unlikely to represent the primary molecular target of KP772, as no significant interaction or damage of DNA was detectable both in vitro and in living cells. Moreover, we found no evidence for induction of radical species. In contrast, KP772 potently inhibited DNA synthesis paralleled by a massive block of cell cycle in G0/G1 phase and a selective decrease of cyclin B1. Although treatment with KP772 induced expression of p53 and p21Waf1, transfection of wild-type p53 into knock out cells only marginally enhanced the cytostatic activity of KP772. In vivo, the anticancer activity of KP772 against human DLD-1 colon carcinoma xenografts was comparable to that of cisplatin and methotrexate at doses not causing significant adverse effects. With regard to toxicity, the LD50 and no-observed-adverse-effect levels (NOAEL) of KP772 in Sprague-Dawley rats were 21.6 and 7.5 mg/kg, in outbred albino mice 62 and 10 mg/kg, respectively. In summary, KP772 exerts anticancer activity via potent induction of cell cycle arrest and/or apoptosis and has promising in vivo anticancer activity against a human colon cancer xenograft. Together, these data suggest further development of KP772 as a new anticancer metal-drug. PMID- 16343447 TI - Conformational induction is the key process for activation of the AT1 receptor. AB - It is currently unclear whether activation of the AT1 receptor by agonists involves conformational selection or induction. We evaluated the pharmacological properties of wild type and N111G CAM human AT1 receptors stably expressed in HEK293 cells. Although [Sar1]-Ang II and Ang IV were full agonists at both receptors, the potency of Ang IV was 280-fold lower at the wild type receptor. [Sar1, Ile8]-Ang II was only a full agonist at the N111G CAM AT1 receptor. [Sar1] Ang II and [Sar1, Ile8]-Ang II displayed similar high affinity binding to both receptors. In contrast, Ang IV displayed low affinity binding to the wild type and high affinity binding to the N111G CAM AT1 receptor. Based on these observations we provide strong evidence that conformational induction is the key process for activation of the AT1 receptor. Only by the creation of CAMs can conformational selection be envisaged to take place. PMID- 16343448 TI - Ontogeny of the dopamine innervation in the nucleus accumbens of gerbils. AB - The postnatal maturation of immunohistochemically stained dopamine (DA) fibres was quantitatively examined in the core and shell subareas of the nucleus accumbens (NAC) of gerbils. Animals of different ages, ranging from juvenile [postnatal day (PD) 14, 30] to adolescent (PD70), adult (PD90, PD180, PD360) and ageing (PD540, PD720) were analysed. The timescale of the maturation of the accumbal DA innervation was regionally different, probably due to the different origin of DA fibres in the mesencephalon. Both the accumbal core, with DA afferents arising from the lateral ventral tegmental area (VTA) and the substantia nigra pars compacta, as well as the accumbal shell, with DA afferents arising from the medial VTA, show moderate DA fibre densities at PD14. The core displayed a significant decrease of the DA fibre density up to PD30 and a subsequent significant increase between PD70 and 90, whereas the shell solely showed an augmentation of the DA innervation between PD70 and 90. Our data suggest that the different maturation of the DA innervation in core and shell might reflect differences in the development of motor and limbic functions, mediated by the nigrostriate and the mesolimbic system, respectively. PMID- 16343449 TI - Reduction of GFAP induced by long dark rearing is not restricted to visual cortex. AB - A key component of the astrocyte cytoskeleton is the glial fibrillary acidic protein (GFAP), which plays an essential role in neuron/astrocyte interactions. Environmental conditioning, such as visual experience manipulation, can affect neuronal and/or glial plasticity in specific brain areas. Previous work from our laboratory showed that short light deprivation throughout the period of GFAP maturation does not influence the expression profile of GFAP in mouse visual cortex; however, it was strong enough to affect neuronal phenotype. It was suggested that visual experience controls the maturation of the neuronal circuitry in this brain area. Therefore, to see whether the modifications of neuronal activity induced by light deprivation affect the maintenance of normal astrocytic phenotype, the dark rearing protocol was extended until the adult life. GFAP-immunoreactive cells were dramatically affected, showing an 80% decrease in number. In addition, GFAP protein level exhibited a 50% reduction, while its mRNA remained unaffected. Besides the visual cortex, two other areas of the brain not directly involved in vision, the hippocampus and the motor cortex, were chosen as internal controls. Unexpectedly, also in these areas, astrocytes were affected by light deprivation. The present results show that lack of visual experience for long periods of time deeply affects glial phenotype not only in visual areas but also in brain regions not directly involved in sensory processing. PMID- 16343450 TI - Neural correlates of human wayfinding in stroke patients. AB - Wayfinding is a complex cognitive function involving different types of information, such as knowledge about landmarks and direction information. This variety of processes suggest that multiple neural mechanisms are involved, e.g., the hippocampal system, the posterior parietal and temporal cortical areas. Although patient studies and imaging studies have given important insights in the exact neural circuitry underlying wayfinding, many controversies remain. Therefore, the current study sets out to further examine the neuroanatomical correlates of wayfinding in a sample of 31 stroke patients with unilateral lesions, tested with a series of different wayfinding tasks, including landmark recognition, landmark ordering, route reversal and route drawing. For all patients, the exact location of their lesion was determined using CT or MRI scans. Based on existing literature, a number of relevant brain areas were demarcated, after which the extent of damage to these areas was determined for each patient separately. Performance on the landmark recognition task was impaired by damage to the right hippocampal formation, whereas a weak correlation was found between damage to the dorsolateral prefrontal cortex and processing the order of the landmarks. Several brain areas were found to be involved in retracing a route from the end to the beginning, including the right hippocampal formation, the right posterior parietal cortex, the right dorsolateral prefrontal cortex and the right temporal lobe. Finally, damage to the right temporal lobe impaired the ability to draw the route. PMID- 16343451 TI - Medial parabrachial nucleus neurons modulate d-fenfluramine-induced anorexia through 5HT2C receptors. AB - We previously reported that lesions of the medial parabrachial nucleus (PBN) enhanced d-fenfluramine (DFEN)-induced anorexia; a finding that suggests these lesions may potentiate the release of serotonin (5HT) or increase the postsynaptic action of 5HT. In the present study, we used SB 206553 (a 5HT2B/2C receptor antagonist) or m-CPP (a 5HT2C/1B receptor agonist) in a standard behavioral procedure (deprivation-induced feeding) to further explore the role of the medial PBN in drug-induced anorexia. In Experiment 1, DFEN (0 or 1.0 mg/kg) was given alone or in combination with SB 206553 (2.0 or 5.0 mg/kg). In Experiment 2, we investigated the food-suppressive effects of m-CPP (0.5, 1.0 or 2.0 mg/kg). The results of Experiment 1 show that SB 206553, while having no influence on the performance of control subjects, attenuated (2.0 mg/kg) or abolished (5 mg/kg) the potentiating effect of the lesions on DFEN-induced anorexia. In Experiment 2, m-CPP induced a suppression of food intake in nonlesioned animals that was significantly potentiated in rats with medial PBN lesions. These results are consistent with the hypothesis that medial PBN neurons mediate anorexia through 5HT2C receptors. PMID- 16343452 TI - Behavioral changes resulting from the administration of cycloheximide in the pilocarpine model of epilepsy. AB - Cycloheximide influences synaptic reorganization resulting from pilocarpine induced status epilepticus (SE). To investigate the possible behavioral consequences of this effect, we subjected animals to pilocarpine-induced SE either in the absence (Pilo group) or presence of cycloheximide (Chx group). Animals were further divided regarding the occurrence of spontaneous recurrent seizures (SRS). Two months after SE induction animals were exposed to different behavioral tests. Age-matched naive animals were used as controls. All epileptic groups showed a significantly diminished freezing time in contextual and tone fear conditioning, performed poorly in the Morris water maze and present less seconds in immobility position as compared to controls. Only Pilo animals explored more extensively the open arms of the elevated plus maze and showed increased in horizontal exploratory activity in the open field as compared to controls. With the exception of Pilo animals without recorded SRS, all other groups had extensive tissue shrinkage in central nucleus of the amygdala as compared to controls. Cycloheximide-treated animals differed from Pilo animals in the extent of hilar loss and supragranular mossy fiber sprouting as well as tissue shrinkage in the dorsal hippocampus. Despite the histological differences seen in the dorsal hippocampus between experimental groups, no differences were encountered in the cognitive tests used to evaluate dorsal hippocampal function. The encountered histological differences between Chx and Pilo animals, however, might underlie the different emotional responses between the two groups. PMID- 16343459 TI - Fast oxidation of thioglycosides to glycosyl sulfones using KMnO4/CuSO4*5H2O under neutral reaction conditions. AB - A rapid oxidation of thioglycosides to glycosyl sulfones has been achieved using a combination of KMnO4 and CuSO4*5H2O in acetonitrile and water. This reaction protocol has many advantages compared to other methods available for this transformation, including compatibility with acid and base labile functional groups used for the protection of carbohydrates, high yields, fast reaction times, and moderate reaction temperatures. The yields obtained were excellent in all cases. PMID- 16343460 TI - Novel derivatives of chitosan and their antifungal activities in vitro. AB - Three kinds of Schiff bases of carboxymethyl chitosan (CMCTS) were prepared, and their antifungal activities were assessed according to Jasso de Rodriguez's method. The results indicated that 2-(2-hydroxybenzylideneamino)-6 carboxymethylchitosan (HNCMCTS) and 2-(5-chloro-2-hydroxybenzylideneamino)-6 carboxymethylchitosan (HCCMCTS) had better inhibitory effects than those of chitosan or CMCTS against Fusarium oxysporium f. sp. vasinfectum, Alternaria solani, and Valsa mali. PMID- 16343461 TI - Highly stereoselective synthesis and structural characterization of new amino sugar derivatives. AB - 2-C-Nitroalkyl-1,4:3,6-dianhydromannitols were synthesized via a Henry reaction of nitroalkyls with 1,4:3,6-dianhydrofructose. Catalytic hydrogenation then afforded the corresponding vicinal amino alcohols. Oximation of 1,4:3,6 dianhydrofructose with hydroxylamine, followed by hydrogenation, gave 2-amino 1,4:3,6-dianhydro-2-deoxymannitol. All compounds were elucidated by their HRMS, 1H NMR, 13C NMR, and IR spectra. The absolute configurations of the amino sugar derivatives were confirmed by single-crystal X-ray analysis or NOESY spectral studies. The possible mechanism for hydrogenation of the nitro 2-C-nitroalkyl sugar is proposed. The conformations of the fused furan rings of nitro and amino sugar derivatives are presented. PMID- 16343462 TI - Chemo-enzymatic synthesis of eel calcitonin glycosylated at two sites with the same and different carbohydrate structures. AB - Naturally occurring glycopeptides and glycoproteins usually contain more than one glycosylation site, and the structure of the carbohydrate attached is often different from site to site. Therefore, synthetic methods for preparing peptides and proteins that are glycosylated at multiple sites, possibly with different carbohydrate structures, are needed. Here, we report a chemo-enzymatic approach for accomplishing this. Complex-type oligosaccharides were introduced to the calcitonin derivatives that contained two N-acetyl-D-glucosamine (GlcNAc) residues at different sites by treatment with Mucor hiemalis endo-beta-N acetylglucosaminidase. Using this enzymatic transglycosylation reaction, three glycopeptides were produced, a calcitonin derivative with the same complex-type carbohydrate at two sites, and two calcitonin derivatives each with one complex type carbohydrate and one GlcNAc. Starting from the derivatives with one complex type carbohydrate and one GlcNAc, a high-mannose-type oligosaccharide was successfully transferred to the remaining GlcNAc using another endo-beta-N acetylglucosaminidase from Arthrobacter protophormiae. Thus, we were able to obtain glycopeptides containing not only two complex-type carbohydrates, but also both complex and high-mannose-type oligosaccharides in a single molecule. Using the resultant glycosylated calcitonin derivatives, the effects of di-N glycosylation on the structure and the activity of calcitonin were studied. The effect appeared to be predictable from the results of mono-N-glycosylated calcitonin derivatives. PMID- 16343463 TI - A lichenase-like family 12 endo-(1-->4)-beta-glucanase from Aspergillus japonicus: study of the substrate specificity and mode of action on beta-glucans in comparison with other glycoside hydrolases. AB - Using anion-exchange chromatography on Source 15Q followed by hydrophobic interaction chromatography on Source 15 Isopropyl, a lichenase-like endo-(1-->4) beta-glucanase (BG, 28kDa, pI4.1) was isolated from a culture filtrate of Aspergillus japonicus. The enzyme was highly active against barley beta-glucan and lichenan (263 and 267 U/mg protein) and had much lower activity toward carboxymethylcellulose (3.9 U/mg). The mode of action of the BG on barley beta glucan and lichenan was studied in comparison with that of Bacillus subtilis lichenase and endo-(1-->4)-beta-glucanases (EG I, II, and III) of Trichoderma reesei. The BG behaved very similar to the bacterial lichenase, except the tri- and tetrasaccharides formed as the end products of beta-glucan hydrolysis with the BG contained the beta-(1-->3)-glucoside linkage at the non-reducing end, while the lichenase-derived oligosaccharides had the beta-(1-->3)-linkage at the reducing end. The BG was characterized by a high amino acid sequence identity to the EG of Aspergillus kawachii (UniProt entry Q12679) from a family 12 of glycoside hydrolases (96% in 162 identified aa residues out of total 223 residues) and also showed lower sequence similarity to the EglA of Aspergillus niger (O74705). PMID- 16343464 TI - Structural analysis of the lipooligosaccharide-derived oligosaccharide of Histophilus somni (Haemophilus somnus) strain 8025. AB - Previous structural studies in our laboratory on lipooligosaccharide (LOS) inner core oligosaccharide (OS) had identified structures from several strains of Histophilus (Haemophilus) somni (738, 2336, 1P, 129Pt). Recently a type strain 8025 was proposed for this species and we therefore sought to determine the core OS structure of this H. somni strain. Core OS was isolated by standard methods from Westphal purified LOS. Structural information was established by a combination of monosaccharide and methylation analyses, NMR spectroscopy and mass spectrometry. The following structure for the core OS was determined on the basis of the combined data from these experiments: [carbohydrates: see text]. The structure determined contains aspects of other Histophilus somni core OS structures, such as the beta-Gal attached at the 2-position of Hep II (2336), PEtn only at the 6-position of Hep II (738, 129Pt) and a lactose extension from Hep I (1P). Since genetic manipulation has been achieved with this strain, the identification of the core OS structure will enable experiments designed to identify the role of glycosyltransferases involved in LOS biosynthesis. PMID- 16343465 TI - Pd(II)-catalysed and Hg(II)-co-catalysed oxidation of D-glucose and D-fructose by N-bromoacetamide in the presence of perchloric acid: a kinetic and mechanistic study. AB - The kinetics of Pd(II)-catalysed and Hg(II)-co-catalysed oxidation of D-glucose (Glc) and D-fructose (Fru) by N-bromoacetamide (NBA) in the presence of perchloric acid using mercury(II) acetate as a scavenger for Br- ions have been studied. The results show first-order kinetics with respect to NBA at low concentrations, tending to zero order at high concentrations. First-order kinetics with respect to Pd(II) and inverse fractional order in Cl- ions throughout their variation have also been noted. The observed direct proportionality between the first-order rate constant (k1) and the reducing sugar concentration shows departure from the straight line only at very higher concentration of sugar. Addition of acetamide (NHA) decreases the first-order rate constant while the oxidation rate is not influenced by the change in the ionic strength (mu) of the medium. Variation of [Hg(OAc)2] shows a positive effect on the rate of reaction. The observed negative effect in H+ at lower concentrations tends to an insignificant effect at its higher concentrations. The first-order rate constant decreases with an increase in the dielectric constant of the medium. The various activation parameters have also been evaluated. The products of the reactions were identified as arabinonic acid and formic acid for both the hexoses. A plausible mechanism involving HOBr as the reactive oxidising species, Hg(II) as co-catalyst, and [PdCl3.S]-1 as the reactive Pd(II)-sugar complex in the rate-controlling step is proposed. PMID- 16343466 TI - Novel bioactive maloyl glucans from aloe vera gel: isolation, structure elucidation and in vitro bioassays. AB - In this study, three novel maloyl glucans were isolated at temperatures below 15 degrees C from aloe vera gel (Aloe barbadensis Miller). These compounds were characterized using NMR spectroscopy, ESIMS, MALDITOF-MS and capillary electrophoresis. The compounds were characterized as 6-O-(1-L-maloyl)-alpha-,beta D-Glcp (veracylglucan A), alpha-D-Glcp-(1-->4)-6-O-(1-L-maloyl)-alpha,-beta,-D Glcp (veracylglucan B) and alpha-D-Glcp-(1-->4)-tetra-[6-O-(1-L-maloyl)-alpha-D Glcp-(1-->4)]-6-O-(1-L-maloyl)-alpha,-beta-D-Glcp (veracylglucan C). These unusual malic acid acylated carbohydrates were then tested in vitro for effects on cell proliferation and gene expression of proinflammatory cytokines, IL-6, IL 8 and ICAM-1, using RT-PCR. Veracylglucan B demonstrated potent anti-inflammatory and anti-proliferative effects, while Veracylglucan C, on the other hand, exhibited significant cell proliferative and anti-inflammatory activities. Veracylglucan A could only be isolated in smaller quantities, and it proved to be very unstable. Thus no biological effects could be observed in this respect. The in vitro bioassays also indicated that Veracylglucan B and C are antagonistic and competitive in their effects on cell proliferation. The results of this work represent a major step forward in the research on aloe vera gel. This is the first time that two fully chemically characterized compounds are shown to be responsible for known biological activities of aloe vera gel. PMID- 16343467 TI - Chemokines induce matrix metalloproteinase-2 through activation of epidermal growth factor receptor in arterial smooth muscle cells. AB - OBJECTIVE: Matrix metalloproteinases (MMP) are critical to smooth muscle cell (SMC) migration in vivo. MMP-2 dysregulation has been implicated in the pathogenesis of abnormal arterial remodeling, aneurysm formation, and atherosclerotic plaque structure and stability. The chemokine receptors CCR3 and CXCR4 are present and functional on SMC and are up-regulated in vascular diseases such as atherosclerosis. We sought to determine a potential mechanism for chemokine receptor-mediated effects on the vasculature by asking whether the chemokines eotaxin (CCL11), the ligand for CCR3, and stromal cell-derived cell factor (SDF-1, CXCL12), the ligand for CXCR4, induce MMP-2 in SMC. Studies were then performed to define the signaling pathways involved. METHODS AND RESULTS: As determined by RT-PCR, Western blotting and zymography, SDF-1 and eotaxin induce MMP-2 mRNA, protein, and activity in SMC. An anti-CCR3 antibody and a CXCR4 antagonist blocked proMMP-2 induction by SDF-1 and eotaxin, the respective ligands for the chemokine receptors CXCR4 and CCR3, suggesting that the inductions by these chemokines are receptor-mediated. Receptor cross-talk between G-protein-coupled receptors (GPCR) and the epidermal growth factor receptor (EGFR) is a method of expanding the GPCRs' signaling repertoire. We demonstrate, for the first time to our knowledge, that in SMC, chemokine induction of proMMP-2 is dependent on activation of the EGFR. Interestingly, by blocking the ligand binding domain of EGFR, we demonstrate that activation of EGFR by SDF-1 and eotaxin occurs through different cellular pathways. CONCLUSION: The pro inflammatory chemokines eotaxin and SDF induce proMMP-2 activation of EGFR through two different pathways. SDF and eotaxin, as regulators of proMMP-2 expression and by engaging in receptor cross-talk, may play critical roles in atherosclerosis, restenosis, and plaque rupture. These ligands and their respective receptors, CXCR4 and CCR3, therefore may serve as future potential therapeutic targets. PMID- 16343468 TI - Insulin: an overall cardiovascular protector? PMID- 16343469 TI - Homocysteine and other biochemical parameters in Type 2 diabetes mellitus with different diabetic duration or diabetic retinopathy. AB - BACKGROUND: Cardiovascular risk-related markers in type 2 diabetes mellitus (DM) have not been well understood. METHODS: Serum and urine samples for biochemical and immunologic analysis were collected from 204 normal subjects and 257 type 2 DM patients, the latter of which were further classified by different diabetic duration with or without retinopathy. RESULTS: Glycosylated hemoglobin A1c, triglyceride, lipase, free fatty acid, albumin creatinine ratio (ACR), lactate dehydrogenase (LDH) and homocysteine were significantly increased in DM patients, whereas high density lipoprotein cholesterol and bilirubin were significantly decreased in DM patients, compared with normal subjects. Lipid profiles, ACR, bilirubin, uric acid, creatine kinase, and hsCRP were not changed in DM patients with different diabetic duration or diabetic retinopathy. Lactate dehydrogenase in DM patients with duration >20 years and homocysteine in patients with duration >10 years was significantly higher than those with duration <5 years. Homocysteine was significantly increased in DM patients with retinopathy, compared with DM patients without retinopathy. CONCLUSION: The increased triglyceride, lipase, free fatty acid, albumin creatinine ratio, lactate dehydrogenase and homocysteine as well as decreased high density lipoprotein cholesterol and anti-oxidative bilirubin in DM patients should be cautious and considered as risks for increasing DM complication. Homocysteine might be associated with longer diabetic duration and microvascular complication of retinopathy in diabetes. PMID- 16343470 TI - Linking inflammation and atherogenesis: Soluble markers identified for the detection of risk factors and for early risk assessment. AB - Increasing evidence has shown that atherogenesis is not only caused by hypercholesterolemia. Several risk factors including abdominal obesity, dyslipidemia, hyperglycemia, bacterial and viral infection, hyperhomocysteinemia have been identified recently, all mediated through inflammation, which can lead to atherosclerosis. Several events have also been identified to be involved in the overall inflammation reaction in the blood vessel which include endothelium dysfunction, expression of adhesion molecules, recruitment of leukocytes to the injured endothelium, migration of monocytes to the arterial intima, and transformation of monocytes to macrophages. In order to facilitate the assessment of early risk for atherogenesis we have made an effort in this review to identify soluble markers that will allow the detection of these risk factors and the identification of associated inflammation events. Since early risks for atherogenesis are largely preventable with dietary modification and lifestyle changes, capable of detecting early risks by monitoring soluble risk markers is conceivably important for asymptomatic individuals to avoid serious or fatal consequences of atherosclerosis. These soluble markers should also be useful for monitoring the effectiveness of intervention and for the identification of therapeutic targets. PMID- 16343472 TI - Bridging the gap between medical and bioinformatics: an ontological case study in colon carcinoma. AB - Ontological principles are needed in order to bridge the gap between medical and biological information in a robust and computable fashion. This is essential in order to draw inferences across the levels of granularity which span medicine and biology, an example of which include the understanding of the roles of tumor markers in the development and progress of carcinoma. Such information integration is also important for the integration of genomics information with the information contained in the electronic patient records in such a way that real time conclusions can be drawn. In this paper, we describe a large multi granular datasource built by using ontological principles and focusing on the case of colon carcinoma. PMID- 16343471 TI - Changes of plasma inflammatory markers after withdrawal of statin therapy in patients with hyperlipidemia. AB - BACKGROUND: Atherosclerosis has been considered to be an inflammatory process. In addition to its lipid-lowering properties, statin has been shown to decrease the concentrations of inflammatory markers resulting in reduction of cardiovascular events. Emerging data suggest that withdrawal of statin might be associated with increased cardiac events. The mechanism for this phenomenon, however, is still unclear. We investigated whether acute termination of statin treatment could result in rebound of inflammatory markers, such as C-reactive protein (CRP) and interleukin-6 (IL-6), in patients with hyperlipidemia. METHODS: Seventeen patients (11 men and 6 women, mean age 51+/-7 years) with hyperlipidemia were given 40 mg/day of pravastatin for 6 weeks. The concentrations of plasma CRP and IL-6 were evaluated before receiving the statin therapy, immediately after 6 weeks of pravastatin therapy, and at days 1, 3 and 7 after withdrawal of pravastatin therapy. The lipid profile was also evaluated at baseline, 6 weeks of therapy, and at day 7 after terminating pravastatin. RESULTS: Pravastatin therapy induced significant reductions in total cholesterol (TC, 6.88+/-0.36 vs. 5.27+/ 0.23 mmol/l, p<0.01), low-density lipoprotein (LDL) cholesterol (4.28+/-0.25 vs. 3.06+/-0.14 mmol/l, p<0.01), CRP (0.28+/-0.16 vs. 0.20+/-0.08 mg/l, p<0.01), and IL-6 (8.4+/-0.6 vs. 6.7+/-0.4 pg/dl, p<0.01). Although the TC and LDL-cholesterol did not change during the 7-day period after withdrawal of pravastatin therapy, the concentrations of CRP and IL-6 increased at day 3 (CRP: 0.20+/-0.08 vs. 0.27+/-0.12 mg/l, and IL-6: 6.7+/-0.4 vs. 7.7+/-0.6 pg/dl, p<0.05 respectively) and at day 7 (CRP: 0.20+/-0.08 vs. 0.30+/-0.14 mg/l, and IL-6: 6.7+/-0.4 vs. 8.7+/-0.8 pg/dl, p<0.01 respectively) after withdrawal of pravastatin therapy. No correlation between increase of CRP as well as IL-6 and small changes of LDL cholesterol concentrations was found after withdrawal of pravastatin therapy at day 7 (r=-0.021 and r=-0.044 respectively, p>0.05 respectively). CONCLUSIONS: 6 weeks after pravastatin therapy could significant modify the lipid profile and decrease the inflammatory markers including CRP and IL-6 in patients with hyperlididemia. Moreover, statin therapy discontinuation could induce a rebound phenomenon of inflammatory response representing an increase in some inflammatory markers, which is independent of changes of lipid parameters. PMID- 16343473 TI - Classification of carotid artery Doppler signals in the early phase of atherosclerosis using complex-valued artificial neural network. AB - In this study, carotid arterial Doppler ultrasound signals were acquired from left carotid arteries of 38 patients and 40 healthy volunteers. The patient group had an established diagnosis of the early phase of atherosclerosis through coronary or aortofemoropopliteal angiographies. Results were classified using complex-valued artificial neural network (CVANN). Principal component analysis (PCA) and fuzzy c-means clustering (FCM) algorithm were used to make a CVANN system more effective. For this aim, before classifying with CVANN, PCA method was used for feature extraction in PCA-CVANN architecture and FCM algorithm was used for data set reduction in FCM-CVANN architecture. Training and test data were selected randomly using 10-fold cross validation. PCA-CVANN and FCM-CVANN architectures classified healthy and unhealthy subjects for training and test data with about 100% correct classification rate. These results shown that PCA CVANN and FCM-CVANN classified Doppler signals successfully. PMID- 16343474 TI - Survival of mint shoot tips after exposure to cryoprotectant solution components. AB - Many plant species can be cryopreserved by treating shoot tips with complex cryoprotectant solutions before rapidly cooling them to liquid nitrogen temperatures. Plant vitrification solution 2 (PVS2), a commonly selected cryoprotectant, can be lethal with extended exposure times. To determine potentially toxic combinations, we have exposed mint shoot tips to one-, two-, three-, and four-component solutions of PVS2 chemicals (30% glycerol, 15% ethylene glycol, 15% dimethyl sulfoxide, and 0.4 M sucrose) at 0 and 22 degrees C. Overall, solution exposures at 22 degrees C were more damaging than exposures at 0 degree C. Solutions with glycerol, particularly in combination with ethylene glycol and dimethyl sulfoxide, were also damaging. Cryoprotectant solutions PGluD (10% PEG8000, 10% glucose, and 10% dimethyl sulfoxide) and PVS3 (50% glycerol, 50% sucrose) were less damaging than PVS2 at 22 degrees C. When plant cryoprotectants are characterized on a toxicological and biophysical basis, less damaging cryoprotectant solutions could be developed. PMID- 16343475 TI - Water transport and IIF parameters for a connective tissue equivalent. AB - Understanding the biophysical processes that govern freezing injury of a tissue equivalent (TE) is an important step in characterizing and improving the cryopreservation of these systems. TEs were formed by entrapping human dermal fibroblasts (HDFs) in collagen or in fibrin gels. Freezing studies were conducted using a Linkam cryostage fitted to an optical microscope allowing observation of the TEs cooled under controlled rates between 5 and 130 degrees C/min. Typically, freezing of cellular systems results in two biophysical processes that are both dependent on the cooling rate: dehydration and/or intracellular ice formation (IIF). Both these processes can potentially be destructive to cells. In this study, the biophysics of freezing cells in collagen and fibrin TEs have been quantified and compared to freezing cells in suspension. Experimental data were fitted in numerical models to extract parameters that governed water permeability, E(Lp) and L(pg), and intracellular ice nucleation, omega(o) and kappa(o). Results indicate that major differences exist between freezing HDFs in suspension and in a tissue equivalent. During freezing, 55% of the HDFs in suspension formed IIF as compared to 100% of HDFs forming IIF in collagen and fibrin TE at a cooling rate of 130 degrees C/min. Also, both the water permeability and the IIF parameters were determined to be higher for HDFs in TEs as compared to cell suspensions. Between the TEs, HDFs in fibrin TE exhibited higher values for the biophysical parameters as compared to HDFs in collagen TE. The observed biophysics seems to indicate that cell-cell and cell-matrix interactions play a major role in ice propagation in TEs. PMID- 16343476 TI - Lymphangiogenesis promotes lens destruction and subsequent lens regeneration in the newt eyeball, and both processes can be accelerated by transplantation of dendritic cells. AB - We examined whether lymphangiogenesis is essential for the process of lens destruction and subsequent remodeling in the newt eye. Lens regeneration was induced by pricking the lens once with a needle through the cornea. The results showed that the formation of the vacuoles which was mediated by lysosomes occurred in the original lens on 8 days after pricking, and histolysis of the lens was induced 24 h later. At that time, new lymphatic vessels appeared in the normally avascular cornea. Immunofluorescence studies revealed the expression of VEGF receptor not only on the cells in the central cornea but also on those in the dorsal iris. Moreover, dendritic cells (DCs) migrated from the peripheral to the central regions in the cornea to engulf the remains of the lens. Next, to determine the extent to which the DCs are important for lens regeneration, we transplanted the DCs that had engulfed the remains of the lens into the eyeball of the normal animals. Interestingly, lens regeneration began in the dorsal iris of eyeballs into which the DCs were transplanted and also in those in which no DCs were transplanted. However, surgical removal of the spleen of the recipient animals prior to transplantation resulted in both a failure of both the VEGFR expression in the dorsal iris and a failure of the novel regeneration. PMID- 16343477 TI - Effect of p75NTR on the regulation of naturally occurring cell death and retinal ganglion cell number in the mouse eye. AB - Neurotrophins induce neural cell survival and differentiation during retinal development and regeneration through the high-affinity tyrosine kinase (Trk) receptors. On the other hand, nerve growth factor (NGF) binding to the low affinity neurotrophin receptor p75 (p75(NTR)) might induce programmed cell death (PCD) in the early phase of retinal development. In the present study, we examined the retinal cell types that experience p75(NTR)-induced PCD and identify them to be postmitotic retinal ganglion cells (RGCs). However, retinal morphology, RGC number, and BrdU-positive cell number in p75(NTR) knockout (KO) mouse were normal after embryonic day 15 (E15). In chick retina, migratory RGCs express p75(NTR), whereas layered RGCs express the high-affinity NGF receptor TrkA, which may switch the pro-apoptotic signaling of p75(NTR) into a neurotrophic one. In contrast to the chick model, migratory RGCs express TrkA, while stratified RGCs express p75(NTR) in mouse retina. However, RGC number in TrkA KO mouse was also normal at birth. We next examined the expression of transforming growth factor beta (TGFbeta) receptor, which modulates chick RGC number in combination with p75(NTR), but was absent in mouse RGCs. p75(NTR) and TrkA seem to be involved in the regulation of mouse RGC number in the early phase of retinal development, but the number may be later adjusted by other molecules. These results suggest the different mechanism of RGC number control between mouse and chick retina. PMID- 16343478 TI - FGF8, Wnt8 and Myf5 are target genes of Tbx6 during anteroposterior specification in Xenopus embryo. AB - The T-box transcription factor Tbx6 is required for somite formation and loss-of function or reduced activity of Tbx6 result in absence of posterior paraxial mesoderm or disorganized somites, but how it is involved in a regulatory hierarchy during Xenopus early development is not clear. We show here that Tbx6 is expressed in the lateral and ventral mesoderm of early gastrula, and it is necessary and sufficient to directly and indirectly regulate the expression of a subset of early mesodermal and endodermal genes. Ectopic expression of Tbx6 inhibits early neuroectodermal gene expression by strongly inducing the expression of posterior mesodermal genes, and expands the mesoderm territory at the expense of neuroectoderm. Conversely, overexpression of a dominant negative Tbx6 mutant in the ventral mesoderm inhibits the expression of several mesodermal genes and results in neural induction in a dose-dependent manner. Using a hormone inducible form of Tbx6, we have identified FGF8, Xwnt8 and XMyf5 as immediate early responsive genes of Tbx6, and the induction of these genes by Tbx6 is independent of Xbra and VegT. These target genes act downstream and mediate the function of Tbx6 in anteroposterior specification. Our results therefore identify a regulatory cascade governed by Tbx6 in the specification of posterior mesoderm during Xenopus early development. PMID- 16343479 TI - KATP channels in mouse spermatogenic cells and sperm, and their role in capacitation. AB - Mammalian sperm must undergo a series of physiological changes after leaving the testis to become competent for fertilization. These changes, collectively known as capacitation, occur in the female reproductive tract where the sperm plasma membrane is modified in terms of its components and ionic permeability. Among other events, mouse sperm capacitation leads to an increase in the intracellular Ca(2+) and pH as well as to a hyperpolarization of the membrane potential. It is well known that ion channels play a crucial role in these events, though the molecular identity of the particular channels involved in capacitation is poorly defined. In the present work, we report the identification and potential functional role of K(ATP) channels in mouse spermatogenic cells and sperm. By using whole-cell patch clamp recordings in mouse spermatogenic cells, we found K(+) inwardly rectifying (K(ir)) currents that are sensitive to Ba(2+), glucose and the sulfonylureas (tolbutamide and glibenclamide) that block K(ATP) channels. The presence of these channels was confirmed using inhibitors of the ATP synthesis and K(ATP) channel activators. Furthermore, RT-PCR assays allowed us to detect transcripts for the K(ATP) subunits SUR1, SUR2, K(ir)6.1 and K(ir)6.2 in total RNA from elongated spermatids. In addition, immunoconfocal microscopy revealed the presence of these K(ATP) subunits in mouse spermatogenic cells and sperm. Notably, incubation of sperm with tolbutamide during capacitation abolished hyperpolarization and significantly decreased the percentage of AR in a dose-dependent fashion. Together, our results provide evidence for the presence of K(ATP) channels in mouse spermatogenic cells and sperm and disclose the contribution of these channels to the capacitation-associated hyperpolarization. PMID- 16343480 TI - The protective effect of prostacyclin on adriamycin-induced apoptosis in rat renal tubular cells. AB - Adriamycin-induced nephrosis in rats is a commonly used experimental model for pharmacological studies of human chronic renal diseases. Adriamycin-induced apoptosis of renal tubular cells has been reported in adriamycin-treated rats. In addition, prostacyclin (PGI(2)) is known to have various protective effects on many kinds of cells. To investigate the protective effect of PGI(2) on cells undergoing adriamycin-induced apoptosis, this study selectively augmented PGI(2) production via adenovirus-mediated transfer of genes for cyclooxygenase-1 (COX-1) and prostacyclin synthase (PGIS) (two key enzymes of PGI(2) synthesis) to renal tubular cells. This PGI(2) overexpression protected rat renal tubular cells from adriamycin-induced apoptosis. Ad-COX-1/PGIS transfection was found to reduce the adriamycin-stimulated activities of caspase-3 and caspase-9, inhibit adriamycin induced release of cytochrome c, elevate the expression of Bcl-x(L), and suppress the activation and translocation of nuclear factor-kappaB (NF-kappaB) in adriamycin-treated renal tubular cells. Our results reveal that selective augmentation of PGI(2) production can protect rat renal tubular cells from adriamycin-induced apoptosis via the NF-kappaB signaling pathway. This implies the therapeutic potential of combined COX-1 and PGIS gene transfer in gene therapy for chronic renal diseases. PMID- 16343481 TI - Anandamide inhibits adhesion and migration of breast cancer cells. AB - The endocannabinoid system regulates cell proliferation in human breast cancer cells. We reasoned that stimulation of cannabinoid CB1 receptors could induce a non-invasive phenotype in breast metastatic cells. In a model of metastatic spreading in vivo, the metabolically stable anandamide analogue, 2-methyl-2'-F anandamide (Met-F-AEA), significantly reduced the number and dimension of metastatic nodes, this effect being antagonized by the selective CB1 antagonist SR141716A. In MDA-MB-231 cells, a highly invasive human breast cancer cell line, and in TSA-E1 cells, a murine breast cancer cell line, Met-F-AEA inhibited adhesion and migration on type IV collagen in vitro without modifying integrin expression: both these effects were antagonized by SR141716A. In order to understand the molecular mechanism involved in these processes, we analyzed the phosphorylation of FAK and Src, two tyrosine kinases involved in migration and adhesion. In Met-F-AEA-treated cells, we observed a decreased tyrosine phosphorylation of both FAK and Src, this effect being attenuated by SR141716A. We propose that CB1 receptor agonists inhibit tumor cell invasion and metastasis by modulating FAK phosphorylation, and that CB1 receptor activation might represent a novel therapeutic strategy to slow down the growth of breast carcinoma and to inhibit its metastatic diffusion in vivo. PMID- 16343482 TI - Coculture with endothelial cells reduces the population of cycling LeX neural precursors but increases that of quiescent cells with a side population phenotype. AB - Neural stem cell proliferation and differentiation are regulated by external cues from their microenvironment. As endothelial cells are closely associated with neural stem cell in brain germinal zones, we investigated whether endothelial cells may interfere with neurogenesis. Neural precursor cells (NPC) from telencephalon of EGFP mouse embryos were cocultured in direct contact with endothelial cells. Endothelial cells did not modify the overall proliferation and apoptosis of neural cells, albeit they transiently delayed spontaneous apoptosis. These effects appeared to be specific to endothelial cells since a decrease in proliferation and a raise in apoptosis were observed in cocultures with fibroblasts. Endothelial cells stimulated the differentiation of NPC into astrocytes and into neurons, whereas they reduced differentiation into oligodendrocytes in comparison to adherent cultures on polyornithine. Determination of NPC clonogenicity and quantification of LeX expression, a marker for NPC, showed that endothelial cells decreased the number of cycling NPC. On the other hand, the presence of endothelial cells increased the number of neural cells having "side population" phenotype, another marker reported on NPC, which we have shown to contain quiescent cells. Thus, we show that endothelial cells may regulate neurogenesis by acting at different level of NPC differentiation, proliferation and quiescence. PMID- 16343483 TI - Bispecific and human disease-related anti-keratin rabbit monoclonal antibodies. AB - Rabbit antibodies may have favorable properties compared to mouse antibodies, including high affinities and better antigen recognition. We used a biochemical and reverse immunologic approach to generate and characterize rabbit anti-phospho keratin and anti-keratin monoclonal antibodies (MAb). Human keratins 8 and 18 (K8/K18) were used as immunogens after isolation from cells pretreated with okadaic acid or pervanadate to promote Ser/Thr or Tyr hyperphosphorylation, respectively. Selected rabbit MAb were tested by immunofluorescence staining, immunoprecipitation, and 2-dimensional gels. Keratin phospho and non-phospho mutants were used for detailed characterization of two unique antibodies. One antibody recognizes a K8 G61-containing epitope, an important epitope given that K8 G61C is a frequent mutation in human liver diseases. This antibody binds K8 that is not phosphorylated on S73, but its binding is ablated by G61 but not S73 mutation. The second antibody is bispecific in that it simultaneously recognizes two epitopes: one phospho (K8 pS431) conformation-independent and one non-phospho conformation-dependent, with both epitopes residing in the K8 tail domain. Therefore, a reverse immunologic and biochemical approach is a viable tool for generating versatile rabbit MAb for a variety of cell biologic applications including the potential identification of physiologic phosphorylation sites. PMID- 16343484 TI - Regulation of lymphangiogenesis--from cell fate determination to vessel remodeling. AB - Lymphatic vessels are important for the maintenance of normal tissue fluid balance, immune surveillance and adsorption of digested fats. During the past decade, the identification of lymphatic-specific markers and growth factors has enabled detailed studies of the lymphatic system, and gain- and loss-of-function experiments have greatly increased our understanding of the mechanisms of normal lymphatic development. Understanding the basic biology has provided novel insights into the pathologic conditions of the lymphatic system that contribute to lymphedema, inflammation or lymphatic metastasis, and opened possibilities for the development of better therapeutic strategies. Here we review the current knowledge about the molecular mechanisms regulating the development of the lymphatic vasculature; of the differentiation of lymphatic endothelial cells, of the regulation of the growth of lymphatic vessels, and of remodeling of the vasculature into a network consisting of lymphatic capillaries and collecting lymphatic vessels. Furthermore, we will discuss the molecular mechanisms involved in the pathological conditions of the lymphatic vessels. PMID- 16343485 TI - Postischemic infusion of adrenomedullin protects against ischemic stroke by inhibiting apoptosis and promoting angiogenesis. AB - Adrenomedullin (AM) is a peptide hormone widely distributed in the central nervous system. Our previous study showed that AM gene delivery immediately after middle cerebral artery occlusion (MCAO) protected against cerebral ischemia/reperfusion (I/R) injury by promoting glial cell survival and migration. In the present study, we investigated the effect of delayed AM peptide infusion on ischemic brain injury at 24 h after MCAO. AM infusion significantly reduced neurological deficit scores at days 2, 4, and 8 after cerebral I/R. AM reduced cerebral infarct size at 8 and 15 days after surgery as determined by quantitative analysis. Double staining showed that AM infusion reduced TUNEL positive apoptotic cells in both neurons and glial cells, as well as reduced caspase-3 activity in the ischemic area of the brain. In addition, AM treatment increased capillary density in the ischemic region at 15 days after I/R injury. Parallel studies revealed that AM treatment enhanced the proliferation of cultured endothelial cells as measured by both (3)H-thymidine incorporation and in situ BrdU labeling. Both in vitro and in vivo AM effects were blocked by calcitonin gene-related peptide (8-37), an AM receptor antagonist. Moreover, AM's effects were associated with increased cerebral nitric oxide (NO) levels, as well as decreased NAD(P)H oxidase activities and superoxide anion production. These results indicate that a continuous supply of exogenous AM peptide protects against I/R injury by improving the survival of neuronal and glial cells, and promoting angiogenesis through elevated NO formation and suppression of oxidative stress. PMID- 16343486 TI - A novel ATP/ADP hydrolysis activity of hyperthermostable group II chaperonin in the presence of cobalt or manganese ion. AB - A novel ATPase activity that was strongly activated in the presence of either cobalt or manganese ion was discovered in the chaperonin from hyperthermophilic Pyrococcus furiosus (Pfu-cpn). Surprisingly, a significant ADPase activity was also detected under the same conditions. A more extensive search revealed similar nucleotide hydrolysis activities in other thermostable chaperonins. Chaperonin activity, i.e., thermal stabilization and refolding of malate dehydrogenase from the guanidine-hydrochloride unfolded state were also detected for Pfu-cpn under the same conditions. We propose that the novel cobalt/manganese-dependent ATP/ADPase activity may be a common trait of various thermostable chaperonins. PMID- 16343487 TI - Identification of novel phospholipid binding proteins in Saccharomyces cerevisiae. AB - A proteomics approach was used to search for novel phospholipid binding proteins in Saccharomyces cerevisiae. Phospholipids were immobilized on a solid support and the lipids were probed with soluble yeast protein extracts. From this, the phosphatidic acid binding proteins were eluted and identified by mass spectrometry. Thirteen proteins were identified and 11 of these were previously unknown lipid binding proteins. The protein-lipid interactions identified would not have been predicted using bioinformatics approaches as none possessed a known lipid binding motif. A subset of the identified proteins was purified to homogeneity and determined to directly bind phospholipids immobilized on a solid support or organized into liposomes. This simple approach could be systematically applied to perform an exhaustive screen for soluble lipid binding proteins in S. cerevisiae or other organisms. PMID- 16343488 TI - Cooperative interaction of Zhangfei and ATF4 in transactivation of the cyclic AMP response element. AB - Zhangfei (ZF) is a basic region-leucine zipper protein that has been implicated in herpesvirus infection cycle and related cellular processes. Here we show both in vivo and in vitro data demonstrating that ZF is a novel cellular binding partner of activating transcription factor 4 (ATF4) (or CREB2). We found that ZF competed with ATF4 to form ATF4-ZF heterodimeric complexes through the bZIP regions. ZF enhanced ATF4 binding to the cAMP response element (CRE), and augmented activation of a CRE reporter by ATF4, in response to MEK1 activation. These results suggest an important role of ZF in the MEK1-ATF4 signaling pathway. PMID- 16343489 TI - Heat shock protein 60: identification of specific epitopes for binding to primary macrophages. AB - In the present study, we characterized regions of human heat shock protein (HSP) 60 responsible for binding to primary macrophages. Studies using 20-mer peptides of the HSP60 sequence to compete with HSP60-binding to macrophages from C57BL/6J mice showed that regions aa241-260, aa391-410 and aa461-480 are involved in surface-binding. HSP60 mutants, lacking the N-terminal 137, 243 or 359 amino acids, inhibited HSP60-binding to primary macrophages to different degrees, demonstrating that all three regions are required for optimal binding. Analysis of different pro- and eukaryotic HSP60 species indicated that phylogenetically separate HSP60 species use different binding sites on primary macrophages. PMID- 16343490 TI - Expression of serum amyloid A transcripts in human trophoblast and fetal-derived trophoblast-like choriocarcinoma cells. AB - The placenta comprises a highly specialized trophoblast layer, which arises from the embryo and differentiates during embryonic development to perform specialized functions, e.g., synthesis of pregnancy-associated hormones, growth factors and cytokines. As there is no evidence of maternal acute-phase protein transplacental transfer and trophoblast plays an important role in regulating immune responses at the feto-maternal interface, the expression of acute-phase serum amyloid A (A SAA) was investigated in human first trimester trophoblast and trophoblast-like JAR and Jeg-3 choriocarcinoma cells. We here show expression of cytokine receptors and cytokine-dependent induction of A-SAA in JAR and Jeg-3 cells. While interleukin-1alpha/beta is a major agonist for A-SAA expression in JAR, tumor necrosis factor-alpha is the predominant agonist in Jeg-3. First trimester trophoblast and JAR/Jeg-3 cells further express the human homolog of SAA activating factor-1, a transcription factor involved in cytokine-mediated induction of A-SAA genes. A-SAA1 and A-SAA2 transcripts were increased in first trimester trophoblast during pregnancy weeks 10 and 12 suggesting that A-SAA plays a role during early fetal development. PMID- 16343491 TI - A caldesmon peptide activates smooth muscle via a mechanism similar to ERK mediated phosphorylation. AB - Caldesmon (CaD) is thought to regulate smooth muscle contraction, because it binds actin and inhibits actomyosin interactions. A synthetic actin-binding peptide (GS17C) corresponding to Gly666-Ser682 of chicken gizzard CaD has been shown to induce force development in permeabilized smooth muscle cells. The mechanism of GS17C's action remains unclear, although a structural effect was postulated. By photo-crosslinking and fluorescence quenching experiments with a gizzard CaD fragment (H32K; Met563-Pro771) and its mutants, we showed that GS17C indeed dissociated the C-terminal region of H32K from actin, in a manner similar to extracellular signal-regulated kinase-mediated phosphorylation, thereby reversing the CaD-imposed inhibition and enabling the actomyosin interaction. PMID- 16343492 TI - Ethanol rapidly causes activation of JNK associated with ER stress under inhibition of ADH. AB - Acute ethanol loading causes oxidative stress to activate cell-death signaling via c-Jun NH2-terminal kinase (JNK) in livers. JNK are stimulated under conditions of endoplasmic reticulum (ER) stress which causes programmed cell death. However, no remarked cell death was observed in acute ethanol intoxication. Akt, one of the cell survival protein kinases, may be activated under ethanol loading. The aim of this study was to estimate activation of JNK and ER stress, role of ethanol metabolism on the activation, and association of JNK with Akt under acute ethanol loading using the perfused rat liver system. Activation of JNK or Akt and association of JNK and Akt with JNK interacting protein 1 were estimated by immunoprecipitation and immunoblotting. Expression of 78 kDa glucose-regulated protein (GRP78) mRNA, a biomarker of ER stress, was detected by quantitative real-time RT-PCR. Activations of JNK and Akt were enhanced by co-treatment with ethanol and a classical inhibitor of alcohol dehydrogenase (ADH). Addition of an antioxidant reduced the activation of JNK. Ethanol loading with ADH inhibition causes down-regulation of GRP78 mRNA levels. Therefore, these findings suggest first revelation that inhibition of ethanol metabolism complicates oxidative and ER stresses produced by ethanol. PMID- 16343493 TI - Disrupted fat absorption attenuates obesity induced by a high-fat diet in Clock mutant mice. AB - The Clock gene is a core component of the circadian clock in mammals. We show here that serum levels of triglyceride and free fatty acid were significantly lower in circadian Clock mutant ICR than in wild-type control mice, whereas total cholesterol and glucose levels did not differ. Moreover, an increase in body weight induced by a high-fat diet was attenuated in homozygous Clock mutant mice. We also found that dietary fat absorption was extremely impaired in Clock mutant mice. Circadian expressions of cholecystokinin-A (CCK-A) receptor and lipase mRNAs were damped in the pancreas of Clock mutant mice. We therefore showed that a Clock mutation attenuates obesity induced by a high-fat diet in mice with an ICR background through impaired dietary fat absorption. Our results suggest that circadian clock molecules play an important role in lipid homeostasis in mammals. PMID- 16343494 TI - A perspective on the role of natriuretic peptides in amphibian osmoregulation. AB - The natriuretic peptide (NP) system is a complex family of peptides and receptors that is primarily linked to the maintenance of osmotic and cardiovascular homeostasis. In amphibians, the potential role(s) of NPs is complicated by the range of osmoregulatory strategies found in amphibians, and the different tissues that participate in osmoregulation. Atrial NP, brain NP, and C-type NP have been isolated or cloned from a number of species, which has enabled physiological studies to be performed with homologous peptides. In addition, three types of NP receptors have been cloned and partially characterised. Natriuretic peptides are always potent vasodilators in amphibian blood vessels, and ANP has been shown to increase the permeability of the microcirculation. In the perfused kidney, ANP causes vasodilation, diuresis and natriuresis that are caused by an increased GFR rather than effects in the renal tubules. These data are supported by the presence of ANP receptors only on the glomeruli and renal blood vessels. In the bladder and skin, the function of NPs is enigmatic because physiological analysis of the effects of ANP on bladder and skin function has yielded conflicting data with no clear role for NPs being revealed. Overall, NPs often have no direct effect, but in some studies they have been shown to inhibit the function of AVT. In addition, there is evidence that ANP can inhibit salt retention in amphibians since it can inhibit the ability of adrenocorticotrophic hormone or angiotensin II to stimulate corticosteroid secretion. It is proposed that an important role for cardiac NPs could be in the control of hypervolaemia during periods of rapid rehydration, which occurs in terrestrial amphibians. PMID- 16343495 TI - Interrenal and thyroid development in red drum (Sciaenops ocellatus): effects of nursery environment on larval growth and cortisol concentration during settlement. AB - Red drum settle into shallow seagrass meadows during the larval stage. Day-night cycles in these habitats result in marked diel temperature and dissolved oxygen (DO) cycles, and it is possible that extreme fluctuations influence endocrine development and growth of larvae. Here, we described red drum interrenal and thyroid ontogeny and determine responses to environmental stimuli with special emphasis on settlement to explore possible role of hormones as mediator of directive environmental factors. This study detected an early activation of thyroid and interrenal axis during the yolk-sac phase and a second activation of the thyroid starting at settlement size to the end of the larval period. Whole body l-thyroxine (T4) and 3-5-3'-triiodo-l-thyronine (T3) showed a sharp decline at the juvenile stage. In contrast, cortisol steadily declines during the larval phase to a minimum before the end of the larval period. Older settlement-size larvae exposed to a strong stimulus increased whole body cortisol. In contrast, new settlers showed a minor cortisol rise suggesting changes on stress responsiveness during the ontogeny of the species. Additionally, settlement-size larvae exposed to various environmentally realistic temperature or DO fluctuations showed no difference in growth compared to fish grown under stable conditions (control). However, growth rate was significantly reduced in DO cycled fish with prolonged exposure to hypoxia. No differences were found in whole-body cortisol levels in the reduced growth treatment groups, suggesting that growth retardation was not related to a cortisol-mediated stress response. In moderate DO and temperature treatment groups, cortisol showed wider fluctuations than control groups during the night time that were not related to stress. PMID- 16343496 TI - The predictive value of serum uric acid, roll-over test, and body mass index in pre-eclampsia. PMID- 16343497 TI - Expression of c-erbB-2 oncoprotein and the prognosis of patients with cervical carcinoma. PMID- 16343498 TI - Sublingual misoprostol versus oxytocin infusion to reduce blood loss at cesarean section. AB - OBJECTIVE: To compare the effectiveness of sublingual misoprostol administered immediately after delivery of the neonate at cesarean section, with intravenous oxytocin infusion in prevention of uterine atony and thereby reducing blood loss at cesarean section. METHODS: One hundred women with singleton term pregnancy undergoing elective or emergency lower segment cesarean section under spinal anesthesia were included in this study. They were randomly allocated to receive either misoprostol 400 mug sublingually or intravenous infusion of 20 units of oxytocin soon after delivery of the neonate. The main outcome measures were blood loss at cesarean section, change in hemoglobin levels, need for additional oxytocics and drug related side effects. RESULTS: The mean blood loss estimated was significantly lower in misoprostol group compared to oxytocin group (819 ml versus 974 ml; p = 0.004). The number of women who had blood loss exceeding 500 ml and the change in hemoglobin, however, was comparable between the two groups. There was a need for additional oxytocic therapy in 16% and 18% after use of misoprostol and oxytocin respectively (p = 0.673). The incidence of side effects such as pyrexia, shivering and metallic taste was significantly higher in misoprostol group compared to oxytocin group. CONCLUSION: Sublingual misoprostol appears to be as effective as intravenous infusion of oxytocin in reducing blood loss at cesarean section. However, occurrence of transient side effects such as shivering and pyrexia were noted more frequently with the use of misoprostol. PMID- 16343499 TI - Sickle cell trait and urinary tract infection in pregnancy. PMID- 16343500 TI - Nurses' responses to expert patients: the rhetoric and reality of self-management in long-term conditions: a grounded theory study. AB - BACKGROUND: Against the backdrop in the western world of increasing prevalence of chronic disease, active and informed patients and a policy emphasis on self management, this English study explored health professionals' responses to expert patients. OBJECTIVES: To: DESIGN: A grounded theory approach was utilised with two concurrent data strands. SETTING: A relatively affluent English county including community, primary and secondary care settings. PARTICIPANTS: Via purposeful and theoretical sampling 100 health professionals (nurses, doctors, physiotherapists) and 100 adults affected by chronic disease participated. METHODS: Focus groups, interviews and observation. RESULTS: Nurses were found to be most anxious about expert patients when compared to other professionals, which appeared to be linked with a lack of professional confidence and unfounded fears regarding litigation. However, nurse specialists often provided a negative case for this. As a whole, nurses were most able to meet the emotional needs of patients, but apart from nurse specialists did not articulate this as a skill. CONCLUSION: Apart from nurse specialists the majority of nurses appeared limited in appropriately facilitating self-management. It is suggested that this is linked to an ongoing nursing culture of patient as passive, an over-emphasis on empirical knowledge and a feeling of vulnerability on the nurses' part towards expert patients. The findings also indicate a rhetoric rather than reality of autonomous nursing roles within the chronic disease management agenda. PMID- 16343501 TI - Bone mineral density in relation to body mass index among young women: a prospective cohort study. AB - AIM: To identify important predictors among lifestyle behaviours and physiological factors of bone mineral density (BMD) in relation to body mass index (BMI) among young women over a 2-year period. DESIGN, SAMPLE AND MEASUREMENTS: Data were collected in 1999 and 2001. Healthy young women (n=152) completed a questionnaire. BMD measurements were performed by DEXA in the calcaneus. The women were subdivided into three categories according to baseline BMI. RESULTS: Baseline bodyweight explained 25% of the variability in BMD at follow-up in the BMI<19 category, and high physical activity seemed to hinder BMD development. In the BMI>24 category, a difference in time spent outdoors during winter between baseline and follow-up was the single most important factor for BMD levels. Overweight women with periods of amenorrhoea had lower BMD than overweight women without such periods. CONCLUSIONS: Predictors and lifestyle behaviours associated with BMD are likely to be based on women of normal weight. BMI should be considered when advising on physical activity, since high physical activity seems to impair BMD development among underweight young women, possibly due to energy imbalance. Among overweight women, sleep satisfaction is the greatest predictor associated with BMD change and may indicate better bone formation conditions. Energy balance and sleep quality may be prerequisites of bone health and should be considered in prevention. PMID- 16343502 TI - Proliferation and anti-apoptotic effects of human urotensin II on human endothelial cells. AB - BACKGROUND: Human urotensin II (hU-II) is a potent vasoconstrictor, highly expressed in cardiac tissues and blood vessels. Recent studies indicate that hU II participates in vascular and myocardial remodeling after injury. This study was designed to study the role of hU-II in cell DNA synthesis and apoptosis in human umbilical vein endothelial cells (HUVECs) and underlying intracellular signaling mechanisms. METHODS AND RESULTS: Cultured HUVECs were incubated with hU II (10(-10)-10(-8)M) for 24h. Cell DNA synthesis was examined by 3H thymidine incorporation. Apoptosis was detected by flow cytometry and TUNEL. hU-II increased the 3H thymidine incorporation into DNA in a concentration-dependent manner. hU-II inhibited endothelial apoptosis induced by serum withdrawal (5.74+/ 0.64% versus 13.20+/-1.96%, P<0.01) and TNFalpha (6.76+/-0.70% versus 13.80+/ 1.02%, P<0.01). The data from flow cytometry and TUNEL are consistent. Further studies showed that hU-II caused the phosphorylation of mitogen-activated protein kinasep42/44 (MAPKp42/44) in a concentration-dependent manner and this effect of hU-II was inhibited by pretreatment of cells with the MEK inhibitor (PD98059, 10muM). In addition, the use of PD98059 also attenuated 3H thymidine incorporation and anti-apoptotic effect elicited by hU-II (both P<0.01 versus hU II alone). CONCLUSIONS: Our observations provide evidence that hU-II promotes cell proliferation and inhibits apoptosis in HUVECs, and MAPKp42/44 activation may play a signal transduction role in this process. PMID- 16343503 TI - Functional mutations of the ABCA1 gene in subjects of French-Canadian descent with HDL deficiency. AB - Mutations in the ABCA1 gene cause defective cellular lipid efflux and severe familial HDL deficiency. We examined the prevalence of mutations at the ABCA1 gene in 58 unrelated probands of French-Canadian descent with HDL deficiency (HDL C<5th percentile). A defective cellular cholesterol or phospholipid efflux (<75% and <70% of normal controls, respectively) was identified in 14/58 (24%) of subjects. Using direct sequencing of the ABCA1 gene, we found mutations in 12/58 ( approximately 20%) of subjects. Four probands were previously identified with diverse ABCA1 gene defects. However, we identified a novel frameshift mutation (F1840L, L1869X); a proband was heteroallelic for the N1800H mutation, previously reported in a case of Tangier disease, and a novel missense mutation (Q2210H); a novel variant (G616V), predicted to impart a functional defect in the protein, was also found in another proband. Three probands had the S1731C mutation, while two others had the R1851X and K776N documented mutations, respectively. Taken together, these data suggest that approximately 20% of French-Canadian patients with severe HDL deficiency are associated with a defective ABCA1. Interestingly, in two families studied, mutations in the ABCA1 gene did not segregate with the lipid efflux defect, suggesting that other proteins are involved in the ABCA1 mediated cellular lipid efflux. PMID- 16343504 TI - Autosomal recessive hypercholesterolemia (ARH) and homozygous familial hypercholesterolemia (FH): a phenotypic comparison. AB - Autosomal recessive hypercholesterolemia (ARH) is a rare disorder, due to complete loss of function of an adaptor protein (ARH protein) required for receptor-mediated hepatic uptake of LDL. ARH is a phenocopy of homozygous familial hypercholesterolemia (HoFH) due to mutations in LDL receptor (LDLR) gene; however, previous studies suggested that ARH phenotype is less severe than that of HoFH. To test this hypothesis we compared 42 HoFH and 42 ARH patients. LDLR and ARH genes were analysed by Southern blotting and sequencing. LDLR activity was measured in cultured fibroblasts. In ARH plasma LDL cholestrol (LDL C) level (14.25+/-2.29 mmol/L) was lower than in receptor-negative HoFH (21.38+/ 3.56 mmol/L) but similar to that found in receptor-defective HoFH (15.52+/-2.39 mmol/L). The risk of coronary artery disease (CAD) was 9-fold lower in ARH patients. No ARH patients 60%, mainly triacylglycerol), cod muscle (low lipid content, approximately 1%, mainly phospholipids), cod plasma (lipid content, approximately 2%, mainly lipoprotein complex, high water amount), cod testis (lipid content approximately 3%, high levels of cholesterol), and herring muscle (lipid content approximately 7%). The one-step procedure for extraction/methylation of wet tissues was compared with the traditional procedure of extraction of the lipids by the Folch method (chloroform/methanol, 2:1, v/v), followed by methylation. The two methods gave similar FA profiles. The one-step extraction/methylation procedure gave a higher recovery of the total FA than the traditional procedure. Problems with carry-over peaks of cholesterol from previous samples were avoided by application of extra long GC temperature programs. The cholesterol decomposed to some degree under the preceding methanolysis step, giving several peaks in the chromatograms. The decomposition peaks were identified by mass spectrometry as cholestdienes originating from dehydration of cholesterol, a metylether of cholesterol and a cholesteryl chloride. These cholesterol artefacts can be used for quantitative determination of cholesterol in the samples. Standard samples of cholesterol were determined with high accuracy, (R(2)>0.99), and cholesterol in cod plasma was compared with good agreement (R(2)=0.97) to an enzymatic method. PMID- 16343518 TI - Identification of flavonoids of Rhodiola rosea by liquid chromatography-tandem mass spectrometry. AB - Phenolic compounds from the aerial parts of medicinal plant Rhodiola rosea were identified using LC/MS experiments with time-of-flight and triple quadrupole instruments, providing accurate mass and CID fragmentation data about the compounds. Supercritical fluid extraction (SFE) was used to remove non-polar compounds from the samples, followed by liquid extraction of the flavonoids. Flavonoids were the main constituents in aerial parts of the plant, and no phenylpropanoids were detected. In addition to usual fragment ions providing the size of the attached glycosides in flavonoids, ions due to radical cleavage of glycosides were observed in the negative ion mode with relatively high collision energies. Use of these ions for elucidating the glycosylation site in the aglycone part was evaluated and was found to give some tentative information, but their use in unambiguous identification of unknown flavonoids is not recommended. Fifteen flavonoids, of which 10 were previously unreported from the plant, were identified. PMID- 16343519 TI - Comparison between sample disruption methods and solid-liquid extraction (SLE) to extract phenolic compounds from Ficus carica leaves. AB - Sea sand disruption method (SSDM) and matrix solid phase disruption (MSPD) were compared to solid-liquid extraction (SLE) for extraction of phenolic compounds from the Ficus carica leaves. Statistical treatment, ANOVA-single factor, was used to compare the extraction yields obtained by these methods, and for the majority of the extracted compounds, significantly higher yields were obtained by the solid disruption methods. Both solid disruption methods are faster and ecologically friendly, but the sea sand method was more reproducible (RSD < 5% for most compounds), and was also the least expensive method. Recoveries above 85% were obtained for chlorogenic acid, rutin, and psoralen using the sea sand extraction method. PMID- 16343521 TI - Comparison of nitrogen adsorption at 77 K on non-porous silica and pore wall of MCM-41 materials by means of density functional theory. AB - Nitrogen adsorption on a surface of a non-porous reference material is widely used in the characterization. Traditionally, the enhancement of solid-fluid potential in a porous solid is accounted for by incorporating the surface curvature into the solid-fluid potential of the flat reference surface. However, this calculation procedure has not been justified experimentally. In this paper, we derive the solid-fluid potential of mesoporous MCM-41 solid by using solely the adsorption isotherm of that solid. This solid-fluid potential is then compared with that of the non-porous reference surface. In derivation of the solid-fluid potential for both reference surface and mesoporous MCM-41 silica (diameter ranging from 3 to 6.5 nm) we employ the nonlocal density functional theory developed for amorphous solids. It is found that, to our surprise, the solid-fluid potential of a porous solid is practically the same as that for the reference surface, indicating that there is no enhancement due to surface curvature. This requires further investigations to explain this unusual departure from our conventional wisdom of curvature-induced enhancement. Accepting the curvature-independent solid-fluid potential derived from the non-porous reference surface, we analyze the hysteresis features of a series of MCM-41 samples. PMID- 16343520 TI - Two-dimensional reversed-phase liquid chromatography using two monolithic silica C18 columns and different mobile phase modifiers in the two dimensions. AB - Comprehensive two-dimensional (2D) HPLC in the reversed-phase liquid chromatography (RPLC) mode using C18 silica monolith columns at first dimension (1st-D) (10 cm x 4.6mm I.D.) and second dimension (2nd-D) (5 cm x 4.6mm I.D.) was carried out successfully. A mixture of water and tetrahydrofuran (THF) was used as a mobile phase in the 1st-D separation, and a mixture of water and methanol (CH3OH) in the 2nd-D separation. Sample fractions from 1st-D column were directly loaded into an injection loop of the 2nd-D HPLC equipped with two injector valves for one column. The fractionation time at the 1st-D that was equal to the separation time at the 2nd-D was 45 or 60s. Total peak capacity up to 900 was obtained in about 60 min for the isocratic mode separation of aromatic compounds in this system. Gradient elution mode applied to both 1st-D and 2nd-D separations resulted in shorter separation time and better separation efficiencies than the isocratic mode. It was demonstrated that 2D-HPLC systems employing popular C18 stationary phases with different organic modifiers in mobile phases for each dimension could produce large peak capacity. The different selectivities were provided by the difference in polar interactions between a solute and the organic modifier existing in the stationary phase. PMID- 16343522 TI - Dispersion of organically modified clays within n-alcohols. AB - A method for formation of polymer-clay nanocomposites involves dispersion of the nanometer silicate layers of clays into a solvent, followed by dispersion into polymers. The dispersion of layered silicates within solvents affects the structure and properties of the nanocomposites. We report the dispersion of organically modified clays, used for formation of nanocomposites with organic polymers, within a range of alcohol solvents. Experiments involved stirring a mixture containing approximately 1 wt% of alkylammonium-modified clays in n alcohols with general molecular structure RnOH, where n represents the number of carbons of alkyl chains, varying from 2 to 8. The clays precipitated from the dispersion when RnOH solvents with n<5 were used, however, they formed gels for solvents with n5. The increased dispersion was related to the decrease of polarity and hydrogen bonding force within solvents. X-ray diffraction for the dispersed clays indicated that the interlayer spaces (1.8 nm), formed by regular stacking of the silicate layers, expanded to a maximum of 3.0 nm after treatment with RnOH with n5. The interlayer expansion was due to the intercalation of n alcohol molecules within the interlayer spaces. It is suggested that the alkyl chains of n-alcohols remain parallel to the silicate surface in the intercalate. Preliminary experiments on the influence of these alcohol solvents on the intercalation of polyol (polyether) are also reported. PMID- 16343523 TI - Interaction forces measured between poly(N-isopropylacrylamide) grafted surface and hydrophobic particle. AB - The interaction forces between poly(N-isopropylacrylamide) (PNIPAAm)-grafted surfaces and colloidal particles in an aqueous solution were investigated using an atomic force microscope. Measurements were conducted between smooth silicon wafers on which PNIPAAm was terminally grafted and silica particles hydrophobized with a silanating reagent in an aqueous electrolyte solution under controlled temperature. Below the lower critical solution temperature (LCST) of PNIPAAm, there were large repulsive forces between the surfaces, both on approach and separation of the surfaces. On the other hand, above LCST, attractive forces were observed both in approaching and in separating force curves. When surface hydrophobicity of the particles increased, the maximum attractive force tended to increase. The changes of hydration state of the grafted PNIPAAm chains depending on temperature are considered to greatly alter the interaction force properties. The role of the intermolecular interaction between the PNIPAAm chains and the hydrophobic particles in the interaction forces is discussed. PMID- 16343524 TI - Interaction of small amounts of bovine serum albumin with phospholipid monolayers investigated by surface pressure and atomic force microscopy. AB - The influence of small amounts of bovine serum albumin (BSA) (nM concentration) on the lateral organization of phospholipid monolayers at the air-water interface and transferred onto solid substrates as one-layer Langmuir-Blodgett (LB) films was investigated. The kinetics of adsorption of BSA onto the phospholipid monolayers was monitored with surface pressure isotherms in a Langmuir trough, for the zwitterionic dipalmitoylphosphatidyl ethanolamine (N,N-dimethyl-PE) and the anionic dimyristoylphosphatidic acid (DMPA). A monolayer of N,N-dimethyl-PE or DMPA incorporating BSA was transferred onto a solid substrate using the Langmuir-Blodgett technique. Atomic force microscopy (AFM) images of one-layer LB films displayed protein-phospholipid domains, whose morphology was characterized using dynamic scaling theories to calculate roughness exponents. For DMPA-BSA films the surface is characteristic of self-affine fractals, which may be described with the Kardar-Parisi-Zhang (KPZ) equation. On the other hand, for N,N dimethyl-PE-BSA films, the results indicate a relatively flat surface within the globule. The height profile and the number and size of globules varied with the type of phospholipid. The overall results, from kinetics of adsorption on Langmuir monolayers and surface morphology in LB films, could be interpreted in terms of the higher affinity of BSA to the anionic DMPA than to the zwitterionic N,N-dimethyl-PE. Furthermore, the effects from such small amounts of BSA in the monolayer point to a cooperative response of DMPA and N,N-dimethyl-PE monolayers to the protein. PMID- 16343525 TI - Phase transfer of gold nanoparticles from aqueous to organic solution containing resorcinarene. AB - Citrate-capped gold nanoparticles (NPs) in aqueous solution were transferred directly into the organic solution mesitylene containing C-undecylcalix[4] resorcinarene (C11-resorcinarene). C11-resorcinarene, which has long hydrophobic tails and phenolic hydroxyl groups, acted as both a phase-transfer and a capping agent. The C11-resorcinarene-capped gold particles could be isolated and dispersed in different organic solvents. Optical absorption spectra corresponding to surface plasmon resonance provided a broad band centered at 534 nm for C11 resorcinarene-capped gold NPs in mesitylene. High-resolution transmission electron micrograph images revealed that the average particle diameter of C11 resorcinarene-capped gold NPs is approximately 12 nm. PMID- 16343526 TI - A simplified method for the rapid fluorometric assessment of antibody-dependent cell-mediated cytotoxicity. AB - We demonstrate that the FATAL cytolysis assay can be adapted into a rapid and fluorometric antibody-dependent cellular cytotoxicity assay (RFADCC). The RFADCC relies on double-staining target cells with a membrane dye (PKH-26) and a viability dye (CFSE) prior to the addition of antibody and effector cells. We used the RFADCC to assess dose-dependent and envelope-specific anti-human immunodeficiency virus (HIV) ADCC responses mediated by monoclonal antibody-2G12 and human sera. Using the assay, we also detected early anti-simian immunodeficiency virus (SIV) ADCC responses in rhesus macaques infected with pathogenic SIV(mac251). Importantly, the RFADCC was further useful in monitoring anti-HIV and anti-SIV ADCC responses elicited by immunizing chimpanzees and rhesus macaques with replicating adenovirus-based AIDS vaccine candidates. In comparison to the standard chromium release assay, the RFADCC provides a higher cell killing readout and is advantageous in allowing use of viably frozen as well as fresh effector cells, thus facilitating assay standardization. The RFADCC is therefore a simple, reliable, and highly sensitive method that can be applied to assess the ADCC activity of monoclonal antibodies as well as ADCC responses elicited by HIV or SIV infection or by AIDS vaccine candidates. PMID- 16343527 TI - Protein translocation through the anthrax toxin transmembrane pore is driven by a proton gradient. AB - Protective antigen (PA) from anthrax toxin assembles into a homoheptamer on cell surfaces and forms complexes with the enzymatic components: lethal factor (LF) and edema factor (EF). Endocytic vesicles containing these complexes are acidified, causing the heptamer to transform into a transmembrane pore that chaperones the passage of unfolded LF and EF into the cytosol. We show in planar lipid bilayers that a physiologically relevant proton gradient (DeltapH, where the endosome is acidified relative to the cytosol) is a potent driving force for translocation of LF, EF and the LF amino-terminal domain (LFN) through the PA63 pore. DeltapH-driven translocation occurs even under a negligible membrane potential. We found that acidic endosomal conditions known to destabilize LFN correlate with an increased translocation rate. The hydrophobic heptad of lumen facing Phe427 residues in PA (or phi clamp) drives translocation synergistically under a DeltapH. We propose that a Brownian ratchet mechanism proposed earlier for the phi clamp is cooperatively linked to a protonation-state, DeltapH-driven ratchet acting trans to the phi-clamp site. In a sense, the channel functions as a proton/protein symporter. PMID- 16343528 TI - Translation of Sindbis virus 26S mRNA does not require intact eukariotic initiation factor 4G. AB - The infection of baby hamster kidney (BHK) cells by Sindbis virus gives rise to a drastic inhibition of cellular translation, while under these conditions the synthesis of viral structural proteins directed by the subgenomic 26S mRNA takes place efficiently. Here, the requirement for intact initiation factor eIF4G for the translation of this subgenomic mRNA has been examined. To this end, SV replicons that contain the protease of human immunodeficiency virus type 1 (HIV 1) or the poliovirus 2A(pro) replacing the sequences of SV glycoproteins have been constructed. BHK cells electroporated with the different RNAs synthesize protein C and the corresponding protease at late times. Notably, the proteolysis of eIF4G by both proteases has little effect on the translation of the 26S mRNA. In addition, recombinant viable SVs were engineered that encode HIV-1 PR or poliovirus 2A protease under the control of a duplicated late promoter. Viral protein synthesis at late times of infection by the recombinant viruses is slightly affected in BHK cells that contain proteolysed eIF4G. The translatability of SV genomic 49S mRNA was assayed in BHK cells infected with a recombinant virus that synthesizes luciferase and transfected with a replicon that expresses poliovirus 2Apro. Under conditions where eIF4G has been hydrolysed significantly the translation of genomic SV RNA was deeply inhibited. These findings indicate a different requirement for intact eIF4G in the translation of genomic and subgenomic SV mRNAs. Finally, the translation of the reporter gene that encodes green fluorescent protein, placed under the control of a second duplicate late promoter, is also resistant to the cleavage of eIF4G. In conclusion, despite the presence of a cap structure in the 5' end of the subgenomic SV mRNA, intact eIF4G is not necessary for its translation. PMID- 16343529 TI - Quantitative analysis of deoxynucleotide substitutions in the codon-anticodon helix. AB - The role of 2' hydroxyl groups in the codon-anticodon helix was evaluated by introducing single deoxynucleotides into each of the six positions in the helix and measuring the affinity of tRNA to either the A site or the P site of Escherichia coli 70S ribosomes. In perfect agreement with the X-ray structure of the Thermus thermophilus 30S subunit, A site binding was weaker in five of the six positions but P site binding was unaffected. Since the addition of paromomycin restores A site binding, it appears that the deoxynucleotide substituted complexes are impaired in their ability to promote the ribosomal conformational change that accompanies tRNA binding. PMID- 16343530 TI - Dimerisation and an increase in active site aromatic groups as adaptations to high temperatures: X-ray solution scattering and substrate-bound crystal structures of Rhodothermus marinus endoglucanase Cel12A. AB - Cellulose, a polysaccharide consisting of beta-1,4-linked glucose, is the major component of plant cell walls and consequently one of the most abundant biopolymers on earth. Carbohydrate polymers such as cellulose are molecules with vast diversity in structure and function, and a multiplicity of hydrolases operating in concert are required for depolymerisation. The bacterium Rhodothermus marinus, isolated from shallow water marine hot springs, produces a number of carbohydrate-degrading enzymes including a family 12 cellulase Cel12A. The structure of R.marinus Cel12A in the ligand-free form (at 1.54 angstroms) and structures of RmCel12A after crystals were soaked in cellopentaose for two different lengths of time, have been determined. The shorter soaked complex revealed the conformation of unhydrolysed cellotetraose, while cellopentaose had been degraded more completely during the longer soak. Comparison of these structures with those of mesophilic family 12 cellulases in complex with inhibitors and substrate revealed that RmCel12A has a more extensive aromatic network in the active site cleft which ejects products after hydrolysis. The substrate structure confirms that during hydrolysis by family 12 cellulases glucose does not pass through a (2,5)B conformation. Small-angle X-ray scattering analysis of RmCel12A showed that the enzyme forms a loosely associated antiparallel dimer in solution, which may target the enzyme to the antiparallel polymer strands in cellulose. PMID- 16343531 TI - Macromolecular crowding in the Escherichia coli periplasm maintains alpha synuclein disorder. AB - The natively disordered protein alpha-synuclein is the primary component of Lewy bodies, the cellular hallmark of Parkinson's disease. Most studies of this protein are performed in dilute solution, but its biologically relevant role is performed in the crowded environment inside cells. We addressed the effects of macromolecular crowding on alpha-synuclein by combining NMR data acquired in living Escherichia coli with in vitro NMR data. The crowded environment in the E.coli periplasm prevents a conformational change that is detected at 35 degrees C in dilute solution. This change is associated with an increase in hydrodynamic radius and the formation of secondary structure in the N-terminal 100 amino acid residues. By preventing this temperature-induced conformational change, crowding in the E.coli periplasm stabilizes the disordered monomer. We obtain the same stabilization in vitro upon crowding alpha-synuclein with 300 g/l of bovine serum albumin, indicating that crowding alone is sufficient to stabilize the disordered, monomeric protein. Two disease-associated variants (A30P and A53T) behave in the same way in both dilute solution and in the E.coli periplasm. These data reveal the importance of approaching the effects of macromolecular crowding on a case-by-case basis. Additionally, our work shows that discrete structured protein conformations may not be achieved by alpha-synuclein inside cells, implicating the commonly overlooked aspect of macromolecular crowding as a possible factor in the etiology of Parkinson's disease. PMID- 16343532 TI - Crystal structures of protein phosphatase-1 bound to motuporin and dihydromicrocystin-LA: elucidation of the mechanism of enzyme inhibition by cyanobacterial toxins. AB - The microcystins and nodularins are tumour promoting hepatotoxins that are responsible for global adverse human health effects and wildlife fatalities in countries where drinking water supplies contain cyanobacteria. The toxins function by inhibiting broad specificity Ser/Thr protein phosphatases in the host cells, thereby disrupting signal transduction pathways. A previous crystal structure of a microcystin bound to the catalytic subunit of protein phosphatase 1 (PP-1c) showed distinct changes in the active site region when compared with protein phosphatase-1 structures bound to other toxins. We have elucidated the crystal structures of the cyanotoxins, motuporin (nodularin-V) and dihydromicrocystin-LA bound to human protein phosphatase-1c (gamma isoform). The atomic structures of these complexes reveal the structural basis for inhibition of protein phosphatases by these toxins. Comparisons of the structures of the cyanobacterial toxin:phosphatase complexes explain the biochemical mechanism by which microcystins but not nodularins permanently modify their protein phosphatase targets by covalent addition to an active site cysteine residue. PMID- 16343533 TI - Glycan microarray analysis of the hemagglutinins from modern and pandemic influenza viruses reveals different receptor specificities. AB - Influenza A virus specificity for the host is mediated by the viral surface glycoprotein hemagglutinin (HA), which binds to receptors containing glycans with terminal sialic acids. Avian viruses preferentially bind to alpha2-3-linked sialic acids on receptors of intestinal epithelial cells, whereas human viruses are specific for the alpha2-6 linkage on epithelial cells of the lungs and upper respiratory tract. To define the receptor preferences of a number of human and avian H1 and H3 viruses, including the 1918 H1N1 pandemic strains, their hemagglutinins were analyzed using a recently described glycan array. The array, which contains 200 carbohydrates and glycoproteins, not only revealed clear differentiation of receptor preferences for alpha2-3 and/or alpha2-6 sialic acid linkage, but could also detect fine differences in HA specificity, such as preferences for fucosylation, sulfation and sialylation at positions 2 (Gal) and 3 (GlcNAc, GalNAc) of the terminal trisaccharide. For the two 1918 HA variants, the South Carolina (SC) HA (with Asp190, Asp225) bound exclusively alpha2-6 receptors, while the New York (NY) variant, which differed only by one residue (Gly225), had mixed alpha2-6/alpha2-3 specificity, especially for sulfated oligosaccharides. Only one mutation of the NY variant (Asp190Glu) was sufficient to revert the HA receptor preference to that of classical avian strains. Thus, the species barrier, as defined by the receptor specificity preferences of 1918 human viruses compared to likely avian virus progenitors, can be circumvented by changes at only two positions in the HA receptor binding site. The glycan array thus provides highly detailed profiles of influenza receptor specificity that can be used to map the evolution of new human pathogenic strains, such as the H5N1 avian influenza. PMID- 16343534 TI - Cell-specific regulation of TRBP1 promoter by NF-Y transcription factor in lymphocytes and astrocytes. AB - HIV-1 viral production is restricted intracellularly in astrocytes compared with lymphocytes due to the limited expression of viral structural proteins. The poor translation of HIV-1 mRNA and consequent limited virion production can be restored by overexpression of TRBP proteins in the astrocytoma U251MG cells. TRBP1 and TRBP2 are double-stranded RNA binding proteins that increase HIV-1 gene expression. Both proteins are produced from a single gene that possesses two independent promoters and an alternative first exon. Endogenous expression is restricted in astrocytes due to limited TRBP promoter expression compared to lymphocytes. We examined the transcriptional regulation of TRBP1 and TRBP2 by in vivo genomic footprinting in the lymphocytic Jurkat and in the astrocytic U251MG cells. We identified one AP4 and one AP2-binding site that regulate the TRBP2 promoter in both cell types, and one Sp1 and two CCAAT-binding sites that control TRBP1 expression. Mutations in the TRBP1 promoter modulate its expression specifically in Jurkat and in U251MG. The analysis of the CCAAT-390 site by EMSA and by ChIP demonstrates that NF-Y/CBF transcription factor binds specifically to the promoter in vitro and in vivo. Furthermore, each NF-Y subunit was more highly expressed in the lymphocytic cells, compared to astrocytic cells. An NF-YA trans dominant mutant decreased TRBP1 promoter expression fourfold in Jurkat cells, thus demonstrating the functional importance of NF-Y factors in lymphocytes. These studies suggest that the cell specifity of HIV-1 expression and replication may be regulated, in part, through the control of TRBP1 expression. PMID- 16343535 TI - Crystal structure and structure-based mutational analyses of RNase HIII from Bacillus stearothermophilus: a new type 2 RNase H with TBP-like substrate-binding domain at the N terminus. AB - Ribonuclease HIII (Bst-RNase HIII) from the moderate thermophile Bacillus stearothermophilus is a type 2 RNase H but shows poor amino acid sequence identity with another type 2 RNase H, RNase HII. It is composed of 310 amino acid residues and acts as a monomer. Bst-RNase HIII has a large N-terminal extension with unknown function and a unique active-site motif (DEDE), both of which are characteristics common to RNases HIII. To understand the role of these N-terminal extension and active-site residues, the crystal structure of Bst-RNase HIII was determined in both metal-free and metal-bound forms at 2.1-2.6 angstroms resolutions. According to these structures, Bst-RNase HIII consists of the N terminal domain and C-terminal RNase H domain. The structures of the N and C terminal domains were similar to those of TATA-box binding proteins and archaeal RNases HII, respectively. The steric configurations of the four conserved active site residues were very similar to those of other type 1 and type 2 RNases H. Single Mn and Mg ions were coordinated with Asp97, Glu98, and Asp202, which correspond to Asp10, Glu48, and Asp70 of Escherichia coli RNase HI, respectively. The mutational studies indicated that the replacement of either one of these residues with Ala resulted in a great reduction of the enzymatic activity. Overproduction, purification, and characterization of the Bst-RNase HIII derivatives with N and/or C-terminal truncations indicated that the N-terminal domain and C-terminal helix are involved in substrate binding, but the former contributes to substrate binding more greatly than the latter. PMID- 16343536 TI - Multi-step assembly pathway of the cbb3-type cytochrome c oxidase complex. AB - The cbb3-type cytochrome c oxidases as members of the heme-copper oxidase superfamily are involved in microaerobic respiration in both pathogenic and non pathogenic proteobacteria. The biogenesis of these multisubunit enzymes, encoded by the ccoNOQP operon, depends on the ccoGHIS gene products, which are proposed to be specifically required for co-factor insertion and maturation of cbb3-type cytochrome c oxidases. Here, the assembly of the cbb3-type cytochrome c oxidase from the facultative photosynthetic model organism Rhodobacter capsulatus was investigated using blue-native polyacrylamide gel electrophoresis. This process involves the formation of a stable but inactive 210 kDa sub-complex consisting of the subunits CcoNOQ and the assembly proteins CcoH and CcoS. By recruiting monomeric CcoP, this sub-complex is converted into an active 230 kDa CcoNOQP complex. Formation of these complexes and the stability of the monomeric CcoP are impaired drastically upon deletion of ccoGHIS. In a ccoI deletion strain, the 230 kDa complex was absent, although monomeric CcoP was still detectable. In contrast, neither of the complexes nor the monomeric CcoP was found in a ccoH deletion strain. In the absence of CcoS, the 230 kDa complex was assembled. However, it exhibited no enzymatic activity, suggesting that CcoS might be involved in a late step of biogenesis. Based on these data, we propose that CcoN, CcoO and CcoQ assemble first into an inactive 210 kDa sub-complex, which is stabilized via its interactions with CcoH and CcoS. Binding of CcoP, and probably subsequent dissociation of CcoH and CcoS, then generates the active 230 kDa complex. The insertion of the heme cofactors into the c-type cytochromes CcoP and CcoO precedes sub-complex formation, while the cofactor insertion into CcoN could occur either before or after the 210 kDa sub-complex formation during the assembly of the cbb3-type cytochrome c oxidase. PMID- 16343537 TI - Concordance of residual dipolar couplings, backbone order parameters and crystallographic B-factors for a small alpha/beta protein: a unified picture of high probability, fast atomic motions in proteins. AB - Using ensemble refinement of the third immunoglobulin binding domain (GB3) of streptococcal protein G (a small alpha/beta protein of 56 residues), we demonstrate that backbone (N-H, N-C', Calpha-Halpha, Calpha-C') residual dipolar coupling data in five independent alignment media, generalized order parameters from 15N relaxation data, and B-factors from a high-resolution (1.1A), room temperature crystal structure are entirely consistent with one another within experimental error. The optimal ensemble size representation is between four and eight, as assessed by complete cross-validation of the residual dipolar couplings. Thus, in the case of GB3, all three observables reflect the same low amplitude anisotropic motions arising from fluctuations in backbone phi/psi torsion angles in the picosecond to nanosecond regime in both solution and crystalline environments, yielding a unified picture of fast, high-probability atomic motions in proteins. An understanding of these motions is crucial for understanding the impact of protein dynamics on protein function, since they provide part of the driving force for triggered conformational changes that occur, for example, upon ligand binding, signal transduction and enzyme catalysis. PMID- 16343538 TI - Visualization of the annealing of complementary single-stranded DNA catalyzed by the herpes simplex virus type 1 ICP8 SSB/recombinase. AB - The rate of annealing of long linear complementary single-stranded (ss) DNAs can be increased greatly by certain DNA-binding proteins including the herpes simplex virus type 1 ICP8 SSB/recombinase. Using electron microscopy, we have investigated the DNA-protein structures involved in ICP8-mediated DNA annealing. We show that the formation of superhelical ICP8-ssDNA filaments is required for annealing. Two superhelices interact with each other to form a coiled-coil, which is the intermediate in annealing. In this process, the superhelices likely rotate and translocate relative to each other. Psoralen/UV photocrosslinking studies revealed that meta-stable contacts form at sites of limited sequence homology during the annealing. Partial proteolysis of ICP8 in the protein-ssDNA complexes showed that Mg2+ induces conformational changes in the N-terminal region (amino acid residues 1-305) of ICP8. In addition to Mg2+, Ca2+ and, to a significantly lesser extent, Cu2+ and Mn2+, were found to induce superhelix formation of the ICP8-ssDNA filament and to facilitate annealing. Mechanisms for how the coiled coil structures facilitate annealing are discussed. PMID- 16343539 TI - Single particle reconstructions of the transferrin-transferrin receptor complex obtained with different specimen preparation techniques. AB - The outcome of three-dimensional (3D) reconstructions in single particle electron microscopy (EM) depends on a number of parameters. We have used the well characterized structure of the transferrin (Tf)-transferrin receptor (TfR) complex to study how specimen preparation techniques influence the outcome of single particle EM reconstructions. The Tf-TfR complex is small (290kDa) and of low symmetry (2-fold). Angular reconstitution from images of vitrified specimens does not reliably converge on the correct structure. Random conical tilt reconstructions from negatively stained specimens are reliable, but show variable degrees of artifacts depending on the negative staining protocol. Alignment of class averages from vitrified specimens to a 3D negative stain reference model using FREALIGN largely eliminated artifacts in the resulting 3D maps, but not completely. Our results stress the need for critical evaluation of structures determined by single particle EM. PMID- 16343541 TI - Unassisted membrane insertion as the initial step in DeltapH/Tat-dependent protein transport. AB - In the thylakoid membrane of chloroplasts as well as in the cytoplasmic membrane of bacteria, the DeltapH/Tat-dependent protein transport pathway is responsible for the translocation of folded proteins. Using the chimeric 16/23 protein as model substrate in thylakoid transport experiments, we dissected the transport process into several distinct steps that are characterized by specific integral translocation intermediates. Formation of the early translocation intermediate Ti 1, which still exposes the N and the C terminus to the stroma, is observed with thylakoids pretreated with (i) solutions of chaotropic salts or alkaline pH, (ii) protease, or (iii) antibodies raised against TatA, TatB, or TatC. Membrane insertion takes place even into liposomes, demonstrating that proteinaceous components are not required. This suggests that Tat-dependent transport may be initiated by the unassisted insertion of the substrate into the lipid bilayer, and that interaction with the Tat translocase takes place only in later stages of the process. PMID- 16343540 TI - Crystal structure of Bacillus anthracis ThiI, a tRNA-modifying enzyme containing the predicted RNA-binding THUMP domain. AB - ThiI is an enzyme responsible for the formation of the modified base S(4)U (4 thiouridine) found at position 8 in some prokaryotic tRNAs. This base acts as a sensitive trigger for the response mechanism to UV exposure, providing protection against its damaging effects. We present the crystal structure of Bacillus anthracis ThiI in complex with AMP, revealing an extended tripartite architecture in which an N-terminal ferredoxin-like domain (NFLD) connects the C-terminal catalytic PP-loop pyrophosphatase domain with a THUMP domain, an ancient predicted RNA-binding domain that is widespread in all kingdoms of life. We describe the structure of the THUMP domain, which appears to be unrelated to RNA binding domains of known structure. Mapping the conserved residues of NFLD and the THUMP domain onto the ThiI structure suggests that these domains jointly form the tRNA-binding surface. The inaccessibility of U8 in the canonical L-shaped form of tRNA, and the existence of a glycine-rich linker joining the catalytic and RNA-binding moieties of ThiI suggest that structural changes may occur in both molecules upon binding. PMID- 16343542 TI - A novel Met116Thr mutation in the GDAP1 gene in a Polish family with the axonal recessive Charcot-Marie-Tooth type 4 disease. AB - Mutations in the gene coding for ganglioside-induced differentiation-associated protein-1 (GDAP1), which maps to chromosome 8q21, have been described in families with autosomal recessive Charcot-Marie-Tooth disease (CMT4A). Interestingly, some mutations in the GDAP1 gene have been reported in the demyelinating form of CMT1 disease, whereas others were found in patients with the axonal type of CMT disease. So far, 23 mutations in the GDAP1 gene have been reported in patients of different ethnic origins. In this study we report a novel mutation Met116Thr in the GDAP1 gene identified in a three generation Polish family with axonal CMT4. PMID- 16343543 TI - Neurology in Operation Iraqi Freedom: risk factors for referral, clinical presentations and incidence of disease. AB - Six hundred and sixteen patients were referred for consultation to the author who served as the neurologist on the 252nd Neurosurgical Team in Kuwait in support of Operation Iraqi Freedom between April and October, 2003. Demographic and military data were collected. The cohort of neurologic patients showed significant differences from the total population of the United States Army contemporarily deployed to Operation Iraqi Freedom. Versus the deployed personnel, the neurologic cohort was older in age (p<0.001), had a greater percentage of females (p<0.00001), had an excessive representation for the military rank of sergeant (p<0.00001), with a deficit of other ranks (junior enlisted and officers), and were more likely to soldiers from the Reserves (p<0.00001) and National Guard (p=0.0021) than from the Regular Army. Seven categories of chief complaints and ten categories of diagnoses constituted some 80% of patients. The incidence of neurologic disease was calculated to be 634 per 100,000 people/year. This information provides valuable information for military neurologists concerning their anticipated duties in future deployments and for non-neurologists by focusing their skills in the evaluation of common neurologic presentations, yet further research is needed to optimize the neurologist's role in a field environment. PMID- 16343544 TI - Hypertonic fluid resuscitation from subarachnoid hemorrhage in rats: a comparison between small volume resuscitation and mannitol. AB - OBJECTIVE: Death and severe morbidity after subarachnoid hemorrhage (SAH) are mainly caused by global cerebral ischemia through increased intracranial pressure (ICP) and decreased cerebral blood flow (CBF). We have recently demonstrated neuroprotective effects of small volume resuscitation (7.5% saline in combination with 6% dextran 70) in an animal model of SAH, leading to normalization of increased ICP, reduced morphological damage and improved neurological recovery. In the present study, we compared the concept of small volume resuscitation represented by two clinically licenced hypertonic-hyperoncotic saline solutions with the routinely used hyperosmotic agent-mannitol-and investigated their effects on ICP, CBF, neurological recovery and morphological damage after SAH in rats. METHODS: 60 dextran-resistant Wistar rats were subjected to SAH by an endovascular filament. ICP, MABP (mean arterial blood pressure) and bilateral local CBF were continuously recorded. All animals were randomly assigned to four groups: (I) NaCl 0.9% (4 ml/kg bw), (II) 7.5% NaCl+6% dextran 70 (4 ml/kg bw), (III) 7.2% NaCl+HES 200,000 (4 ml/kg bw) and (IV) 20% mannitol (9.33 ml/kg bw) given 30 min after SAH. Neurological deficits were assessed on days 1, 3 and 7 after SAH. The morphological damage was evaluated on day 7 after SAH. RESULTS: The induction of SAH resulted in an immediate ICP increase to 46.6+/-3.2 mm Hg (mean+/-S.E.M.) and 29.6+/-1.3 (mean+/-S.E.M.) mm Hg 90 min post-SAH. While a treatment with both hypertonic saline solutions (II, III) decreased ICP as well as the 20% mannitol solution, only the group treated with hypertonic saline and dextran 70 (II) showed an increase of ipsilateral CBF for 20 min after the infusion and significantly more surviving neurons in the motorcortex and caudoputamen. Mortality was reduced from 60% (I) and 73% (III and IV), respectively, to 40% in group II. CONCLUSION: Of all hypertonic solutions investigated, small volume resuscitation with NaCl 7.5% in combination with 6% dextran 70 evolved to be most effective in terms of reducing the initial harmful sequelae of SAH, leading to lowered ICP and less morphological damage after SAH in the rat. PMID- 16343545 TI - Stochastic dynamics of metastasis formation. AB - Tumor metastasis accounts for the majority of deaths in cancer patients. The metastatic behavior of cancer cells is promoted by mutations in many genes, including activation of oncogenes such as RAS and MYC. Here, we develop a mathematical framework to analyse the dynamics of mutations enabling cells to metastasize. We consider situations in which one mutation is necessary to confer metastatic ability to the cell. We study different population sizes of the main tumor and different somatic fitness values of metastatic cells. We compare mutations that are positively selected in the main tumor with those that are neutral or negatively selected, but faster at forming metastases. We study whether metastatic potential is the property of all (or the majority of) cells in the main tumor or only the property of a small subset. Our theory shows how to calculate the expected number of metastases that are formed by a tumor. PMID- 16343546 TI - Design of a network with state stability. AB - Designing a network with given functions or reconstruct a network based on its dynamical behavior is an important problem in the study of complex systems. In this paper, we put forward certain principles in constructing a network with state stability. We show that a necessary and sufficient condition to design networks with a global fixed point is that active nodes inhibit inactive nodes, while the latter activate the former directly or indirectly. We also designed networks based on basic modules, where each basic module consists a sub-network, they communicate through the inhibition link from each activator in lower module to the inhibitor of upper module. We found that long activation links, i.e. indirect activation links are important to the formation of convergence trajectory. We believe that these principles may help us to understand the topology of biological networks. PMID- 16343547 TI - Selection in favor of nucleotides G and C diversifies evolution rates and levels of polymorphism at mammalian synonymous sites. AB - The impact of synonymous nucleotide substitutions on fitness in mammals remains controversial. Despite some indications of selective constraint, synonymous sites are often assumed to be neutral, and the rate of their evolution is used as a proxy for mutation rate. We subdivide all sites into four classes in terms of the mutable CpG context, nonCpG, postC, preG, and postCpreG, and compare four-fold synonymous sites and intron sites residing outside transposable elements. The distribution of the rate of evolution across all synonymous sites is trimodal. Rate of evolution at nonCpG synonymous sites, not preceded by C and not followed by G, is approximately 10% below that at such intron sites. In contrast, rate of evolution at postCpreG synonymous sites is approximately 30% above that at such intron sites. Finally, synonymous and intron postC and preG sites evolve at similar rates. The relationship between the levels of polymorphism at the corresponding synonymous and intron sites is very similar to that between their rates of evolution. Within every class, synonymous sites are occupied by G or C much more often than intron sites, whose nucleotide composition is consistent with neutral mutation-drift equilibrium. These patterns suggest that synonymous sites are under weak selection in favor of G and C, with the average coefficient s approximately 0.25/Ne approximately 10(-5), where Ne is the effective population size. Such selection decelerates evolution and reduces variability at sites with symmetric mutation, but has the opposite effects at sites where the favored nucleotides are more mutable. The amino-acid composition of proteins dictates that many synonymous sites are CpGprone, which causes them, on average, to evolve faster and to be more polymorphic than intron sites. An average genotype carries approximately 10(7) suboptimal nucleotides at synonymous sites, implying synergistic epistasis in selection against them. PMID- 16343548 TI - Spontaneous polarization in eukaryotic gradient sensing: a mathematical model based on mutual inhibition of frontness and backness pathways. AB - A key problem of eukaryotic cell motility is the signaling mechanism of chemoattractant gradient sensing. Recent experiments have revealed the molecular correlate of gradient sensing: Frontness molecules, such as PI3P and Rac, localize at the front end of the cell, and backness molecules, such as Rho and myosin II, accumulate at the back of the cell. Importantly, this frontness backness polarization occurs spontaneously even if the cells are exposed to uniform chemoattractant profiles. The spontaneous polarization suggests that the gradient sensing machinery undergoes a Turing bifurcation. This has led to several classical activator-inhibitor and activator-substrate models which identify the frontness molecules with the activator. Conspicuously absent from these models is any accounting of the backness molecules. This stands in sharp contrast to experiments which show that the backness pathways inhibit the frontness pathways. Here, we formulate a model based on the mutually inhibitory interaction between the frontness and backness pathways. The model builds upon the mutual inhibition model proposed by Bourne and coworkers [Xu et al., 2003. Divergent signals and cytoskeletal assemblies regulate self-organizing polarity in neutrophils. Cell 114, 201-214.]. We show that mutual inhibition alone, without the help of any positive feedback (autocatalysis), can trigger spontaneous polarization of the frontness and backness pathways. The spatial distribution of the frontness and backness molecules in response to inhibition and activation of the frontness and backness pathways are consistent with those observed in experiments. Furthermore, depending on the parameter values, the model yields spatial distributions corresponding to chemoattraction (frontness pathways in-phase with the external gradient) and chemorepulsion (frontness pathways out-of-phase with the external gradient). Analysis of the model suggests a mechanism for the chemorepulsion-to-chemoattraction transition observed in neurons. PMID- 16343549 TI - Erythropoietin prevents the increase in blood-brain barrier permeability during pentylentetrazol induced seizures. AB - Recombinant human erythropoietin (r-Hu EPO) has been shown to exert neuroprotection in ischemic, excitotoxicity, trauma, convulsions and neurodegenerative disorders. Blood-brain barrier (BBB) leakage plays a role in the pathogenesis of many pathological states of the brain including neurodegenerative disorders. This study aimed to investigate the effects of r-Hu EPO on BBB integrity in pentylentetrazol (PTZ) induced seizures in rats. Seizures were observed and evaluated regard to latency and intensity for an hour. Macroscopical and spectrophotometrical measurement of Evans Blue (EB) leakage were observed for BBB integrity. r-Hu EPO was given intraperitoneally 24 h prior to seizure induction. Total seizure duration of 720+/-50 s after single PTZ administration (80 mg/kg i.p.) was declined to 190+/-40 s in r-Hu EPO pretreatment. A typical BBB breakdown pattern (i.e. staining in cerebellum, cerebral cortex, midbrain, hippocampus, thalamus and corpus striatum) was observed in rat brains with PTZ induced seizures; whereas, EPO pretreatment confined BBB leakage to cerebellum and cortical areas, and lessened the intensity of tonic-clonic seizures observed in PTZ seizures. The protective effect of r-Hu EPO on BBB permeability in seizures is a new and original finding. The protective action of r-Hu EPO in seizures and some of CNS pathologies warrant further investigations. PMID- 16343550 TI - Angiotensin II stimulates intercellular adhesion molecule-1 via an AT1 receptor/nuclear factor-kappaB pathway in brain microvascular endothelial cells. AB - Microvascular changes in the brain are significant causes of cerebral edema and ischemia injury. A number of studies suggest that angiotensin (Ang) II may be involved in the initiation and regulation of processes occurring in brain ischemia. We recently reported that Ang II injures brain microvascular endothelial cells (BMEC) partially via stimulating intercellular adhesion molecule-1 (ICAM-1) expression. However, the signaling cascade leading to Ang II induced ICAM-1 expression in BMEC was unclear. The present study tested the hypothesis that Ang II induces ICAM-1 expression via an AT1 receptor/nuclear factor-kappaB (NF-kappaB) pathway in BMEC. Ang II directly stimulated the expression of ICAM-1 mRNA and protein in primary cultured BMEC. Ang II treatment also resulted in the degradation of IkappaBalpha and increase of NF-kappaB p65 subunit in the nucleus as well as the DNA binding activity of nuclear NF-kappaB. These effects were abolished by pretreatment with the selective AT1 receptor antagonists, losartan and compound EXP-2528, or losartan plus the AT2 receptor antagonist PD123319, but not by PD123319 alone. Moreover, there were no significant differences between the losartan and losartan plus PD123319 groups. These findings indicate that Ang II-induced ICAM-1 upregulation in brain microvascular endothelial cells may be mediated via an AT1 receptor/NF-kappaB pathway. PMID- 16343551 TI - Involvement of adenosine A1 and A2A receptors in (-)-linalool-induced antinociception. AB - In recent studies performed in our laboratory we have shown that acute administration of (-)-linalool, the natural occurring enantiomer in essential oils, possesses anti-inflammatory, antihyperalgesic and antinociceptive effects in different animal models. The antihyperalgesic and antinociceptive effects of ( )-linalool have been ascribed to its capacity in stimulating the opioidergic, cholinergic and dopaminergic systems, as well as to its interaction with K+ channels, or to its local anaesthetic activity and/or to the negative modulation of glutamate transmission. Activation of A1 or A2A receptors has been shown to induce antinociceptive effects, and the possible involvement of adenosine in (-) linalool antinociceptive effect, has not been elucidated yet. Therefore, in the present study, we have investigated the effects of 1,3-dipropyl-8 cyclopentylxanthine (DPCPX), a selective adenosine A1 receptor antagonist and the effects of 3,7-dimethyl-1-propargilxanthine (DMPX), a selective adenosine A2A receptor antagonist on the antinociception of (-)-linalool in mice, measured in the hot-plate test. Both DPCPX (0.1 mg/kg; i.p.) and DMPX (0.1 mg/kg; i.p.) pre treatment significantly depressed the antinociceptive effect of (-)-linalool at the highest doses tested. These findings demonstrated that the effect of (-) linalool on pain responses is, at least partially, mediated by the activity of adenosine A1 and A2A receptors. PMID- 16343552 TI - Induction of protein degradation in skeletal muscle by a phorbol ester involves upregulation of the ubiquitin-proteasome proteolytic pathway. AB - Although muscle atrophy is common to a number of disease states there is incomplete knowledge of the cellular mechanisms involved. In this study murine myotubes were treated with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) to evaluate the role of protein kinase C (PKC) as an upstream intermediate in protein degradation. TPA showed a parabolic dose-response curve for the induction of total protein degradation, with an optimal effect at a concentration of 25 nM, and an optimal incubation time of 3 h. Protein degradation was attenuated by co-incubation with the proteasome inhibitor lactacystin (5 microM), suggesting that it was mediated through the ubiquitin-proteasome proteolytic pathway. TPA induced an increased expression and activity of the ubiquitin proteasome pathway, as evidenced by an increased functional activity, and increased expression of the 20S proteasome alpha-subunits, the 19S subunits MSS1 and p42, as well as the ubiquitin conjugating enzyme E2(14k), also with a maximal effect at a concentration of 25 nM and with a 3 h incubation time. There was also a reciprocal decrease in the cellular content of the myofibrillar protein myosin. TPA induced activation of PKC maximally at a concentration of 25 nM and this effect was attenuated by the PKC inhibitor calphostin C (300 nM), as was also total protein degradation. These results suggest that stimulation of PKC in muscle cells initiates protein degradation through the ubiquitin-proteasome pathway. TPA also induced degradation of the inhibitory protein, I-kappaBalpha, and increased nuclear accumulation of nuclear factor-kappaB (NF-kappaB) at the same time and concentrations as those inducing proteasome expression. In addition inhibition of NF-kappaB activation by resveratrol (30 microM) attenuated protein degradation induced by TPA. These results suggest that the induction of proteasome expression by TPA may involve the transcription factor NF-kappaB. PMID- 16343553 TI - Estimation of both the potency and efficacy of alpha7 nAChR agonists from single concentration responses. AB - The assessment of functional properties is a crucial step in the screening of potential new drug candidates. The development of moderate to high throughput electrophysiological recording systems such as OpusXpress (Molecular Devices) has facilitated the process of testing new drugs to a large degree. However, while the simple screening of multiple drugs at a single concentration identifies "hits" and "misses", the generation of full concentration-response studies is still a bottleneck in drug development. The alpha7 nicotinic acetylcholine receptor displays a unique concentration dependence of response kinetics which permits estimates of EC50 and Imax values for experimental drugs to be generated from single-concentration responses. This method is based on the analysis of 13 different concentration-response studies utilizing either human or rat alpha7 nAChR. Each experimental response was first normalized to an ACh control, and then a transformation of the pooled data was generated which, based on the relationship between the net charge and peak current to their respective EC50 values defined the "functional concentration" (the test concentration relative to the EC50 for the given agonist). At low functional concentrations, net charge is large relative to peak current amplitude and at higher functional concentration this relationship reverses. For any single-concentration response, the ratio of net charge to peak current can be used to estimate functional concentration. Efficacy can then be estimated by comparing the observed (net charge) response to the expected value for a full agonist at the estimated functional concentration. This extended analysis, combined with automated recording methods, should greatly increase the efficiency with which promising new drug candidates can be characterized. PMID- 16343554 TI - NAD(P)H oxidase-stimulating activity of serum from type 2 diabetic patients with retinopathy mediates enhanced endothelial expression of E-selectin. AB - Endothelial expression of E-selectin is enhanced in diabetic patients with retinopathy, however, the underlying mechanisms are unclear. Therefore, this study was aimed to determine if endothelial expression of E-selectin is stimulated with serum from type 2 diabetic patients with retinopathy, and whether this process is related to NAD(P)H oxidase-derived oxidative stress. Serum was obtained from type 2 diabetic patients with (T2DR) or without (T2DM) retinopathy, and age-matched non-diabetic healthy person (Control). Serum was added to in vitro-grown human coronary artery endothelial cells (HCAEC), after which E selectin expression, reactive oxygen species (ROS) production, and NAD(P)H oxidase activity were measured. Serum from T2DR induced a significantly higher expression of E-selectin than serum from T2DM and control in association with an enhanced production of ROS in HCAEC. T2DR serum enhanced E-selectin expression in a ROS-dependent manner since this process was significantly attenuated not only by tiron (1 mM), a superoxide scavenger, but also by DPI (10 micromol/L) and apocynin (100 micromol/L), inhibitors of NAD(P)H oxidase. Furthermore, the activity of NADH oxidase was markedly increased by T2DR serum, and this was accompanied by the enhanced membrane translocation of p47phox, a cytosolic subunit of NAD(P)H oxidase. These findings suggest that serum from T2DR induced up-regulation of E-selectin expression in HCAEC, and this process might be dependent on activation of endothelial NADH oxidase via an enhanced membrane translocation of p47phox. PMID- 16343555 TI - Monitoring the Prestige oil spill impacts on some key species of the Northern Iberian shelf. AB - Selected key components of the continental shelf benthic and demersal communities were monitored for the two years following the Prestige oil spill (POS) in order to identify the possible ecological effects of the oil. This work includes the first results regarding changes in abundance, distribution and food habits of hake (Merluccius merluccius), four-spot megrim (Lepidorhombus boscii), Norway lobster (Nephrops norvegicus) and Pandalid shrimp (Plesionika heterocarpus) populations of Galician and Cantabrian Sea shelves following the POS. Significant reductions in the abundance of Norway lobster, Plesionika heterocarpus and four spot megrim were detected in the POS maximum impact area, located over the Galician shelf. Noteworthy recoveries were observed in the 2004 abundance indices of four-spot megrim and Plesionika. On the other hand, no significant effects were detected in the abundance or distribution of hake juveniles even though the tar aggregates were bound by the same oceanographic drift events as the hake recruits were during the winter of 2003 (Navidad current) in different water column layers of the Cantabrian Sea. Feeding patterns of the four species analysed did not present apparent modifications that can be related to the POS. PMID- 16343556 TI - On the intra-host dynamics of HIV-1 infections. AB - An extension of a previously proposed theory for the pathogenesis of AIDS is presented and analyzed using a mathematical modelling approach. This theory is based on the observation that human immunodeficiency virus type 1 (HIV-1) predominantly infects and replicates in (CD4+)-T cells, and that the infection process within an infected individual is characterized by ongoing generation and selection of HIV variants with increasing reproductive capacity. This evolutionary process is considered to be the reason for the gradual loss of immunocompetence and the final destruction of the immune system observed in most patients. The extension presented here incorporates the effect of the permanently increasing susceptibility of (CD4+)-T cell clones, as a result of the evolutionary process. The presented model reproduces and possibly explains a wide variety of findings about the HIV infection process. Numerical results indicate that the effect of the initial dose is minimal, and restricted to the primary phase of infection. According to the model predictions the impact of the HIV evolutionary speed is crucial for the progression to disease. An important progression determinant is the initial infection rate, being a component of the viral reproductive capacity. An influential role in disease progression seems to be played by the initial (CD4+)-T cell count. PMID- 16343557 TI - Quarantine in a multi-species epidemic model with spatial dynamics. AB - Motivation is provided for the development of infectious disease models that incorporate the movement of individuals over a range of spatial scales. A general model is formulated for a disease that can be transmitted between different species and multiple patches, and the behavior of the system is investigated in the case in which the spatial component consists of a ring of patches. The influence of various parameters on the spatial and temporal spread of the disease is studied numerically, with particular focus on the role of quarantine in the form of travel restriction. PMID- 16343558 TI - High frequency induction of mitotic recombination by ionizing radiation in Mlh1 null mouse cells. AB - Mitotic recombination in somatic cells involves crossover events between homologous autosomal chromosomes. This process can convert a cell with a heterozygous deficiency to one with a homozygous deficiency if a mutant allele is present on one of the two homologous autosomes. Thus mitotic recombination often represents the second mutational step in tumor suppressor gene inactivation. In this study we examined the frequency and spectrum of ionizing radiation (IR) induced autosomal mutations affecting Aprt expression in a mouse kidney cell line null for the Mlh1 mismatch repair (MMR) gene. The mutant frequency results demonstrated high frequency induction of mutations by IR exposure and the spectral analysis revealed that most of this response was due to the induction of mitotic recombinational events. High frequency induction of mitotic recombination was not observed in a DNA repair-proficient cell line or in a cell line with an MMR-independent mutator phenotype. These results demonstrate that IR exposure can initiate a process leading to mitotic recombinational events and that MMR function suppresses these events from occurring. PMID- 16343559 TI - Backward inhibition in Parkinson's disease. AB - Parkinson's disease has been associated with executive dysfunction, especially task-switching deficits. One factor contributing to task-switching costs is backward inhibition, as measured by less efficient performance when switching back to a task from which one has recently switched away. This alternating-switch cost is considered to be due to persisting inhibition of elements of the previous task set after a switch. In this study, patients with mild to moderate Parkinson's disease and controls performed three tasks (A-C) in an intermixed fashion. Patients with mild to moderate Parkinson's disease and controls showed equivalent response times. However, the patients made significantly more errors during an alternating switch (i.e., ABA) than did control participants. In contrast, there was no group difference in accuracy in the comparable condition of two consecutive switches between different tasks (i.e., CBA). In addition, accuracy for the two groups was similar for trials in which the task was repeated. These data suggest that Parkinson's disease is associated with either increased backward inhibition, or a reduced ability to overcome this inhibition when reactivating a recently abandoned task set. PMID- 16343560 TI - Mapping the parietal cortex of human and non-human primates. AB - The present essay reviews a series of functional magnetic resonance imaging (fMRI) studies conducted in parallel in humans and awake monkeys, concentrating on the intraparietal sulcus (IPS). MR responses to a range of visual stimuli indicate that the human IPS contains more functional regions along its anterior posterior extent than are known in the monkey. Human IPS includes four motion sensitive regions, ventral IPS (VIPS), parieto-occipital IPS (POIPS), dorsal IPS medial (DIPSM) and dorsal IPS anterior (DIPSA), which are also sensitive to three dimensional structure from motion (3D SFM). On the other hand, the monkey IPS contains only one motion sensitive area (VIP), which is not particularly sensitive to 3D SFM. The human IPS includes four regions sensitive to two dimensional shape and three representations of central vision, while monkey IPS appears to contain only two shape sensitive regions and one central representation. These data support the hypothesis that monkey LIP corresponds to the region of human IPS between DIPSM and POIPS and that a portion of the anterior part of human IPS is evolutionarily new. This additional cortical tissue may provide the capacity for an enhanced visual analysis of moving images necessary for sophisticated control of manipulation and tool handling. PMID- 16343561 TI - Low temperatures during early development influence subsequent maternal and reproductive function in adult female mice. AB - Challenging conditions early in development can have enduring effects into adulthood. The effects of low temperatures were examined on subsequent sex specific morphology (anogenital distance [AGD]), maternal care, and reproductive performance in adult female mice. Dams (F0) were maintained either in (1) standard laboratory room temperatures (21 +/- 2 degrees C) or (2) low temperatures (10 +/- 2 degrees C) throughout gestation. Their progeny (F1) either remained in the temperature condition in which they were conceived or were switched to the other temperature condition at 2 days of age until weaning. Reproductive performance and maternal behaviors were assessed in adulthood. F0 dams that were maintained in low temperatures bore larger litters as compared to F0 animals housed in standard temperatures throughout their pregnancy. In contrast, mean litter size was reduced for all groups of F1 females that experienced low temperatures. Infant mortality was elevated in litters of F1 females that were exposed to low temperatures both before and after birth. Prenatal exposure to low temperatures was associated with reduced responsiveness towards the nursing young and decreased maternal aggression in F1 animals. Prenatally treated F1 females had longer, male-like AGDs on Day 2 following birth compared to animals not subjected to experimental manipulations. Our results indicate that exposure to low temperatures during early development impairs reproductive function and is associated with important fitness costs as evidenced by reduced offspring survival. Our findings also suggest that chronic low temperatures experienced only after birth may have less deleterious effects than exposure to a combination of pre- and postnatal or prenatal treatments alone. PMID- 16343562 TI - Integrated zone comparison polygraph technique accuracy with scoring algorithms. AB - The Integrated Zone Comparison Technique (IZCT) was utilized with computerized polygraph instrumentation as part of a blind study in the detection of deception. Three scoring algorithms: ASIT Poly Suite (Academy for Scientific Investigative Training's Horizontal Scoring and Algorithm for Chart Interpretation), PolyScore 5.5, and the Objective Scoring System (OSS) were assessed in the interpretation of the charts generated. Where "Inconclusives" were excluded, accuracy for the IZCT with all three algorithms was 100%. When "Inconclusives" were counted as errors, overall accuracy for the IZCT with ASIT Poly Suite was 90% and accuracy with PolyScore and the Objective Scoring System was 72%. PMID- 16343563 TI - Modulation of memory with septal injections of morphine and glucose: effects on extracellular glucose levels in the hippocampus. AB - The concentration of glucose in the extracellular fluid (ECF) of the hippocampus decreases substantially during memory testing on a hippocampus-dependent memory task. Administration of exogenous glucose, which enhances task performance, prevents this decrease, suggesting a relationship between hippocampal glucose availability and memory performance. In the present experiment, spontaneous alternation performance and task-related changes in hippocampal ECF glucose were assessed in rats after intraseptal administration of morphine, which impairs memory on a spontaneous alternation task, and after co-administration of intraseptal glucose, which attenuates that impairment. Consistent with previous findings, spontaneous alternation testing resulted in a decrease in hippocampal ECF glucose levels in control rats. However, rats that received intraseptal morphine prior to testing showed memory impairments and an absence of the task related decrease in hippocampal ECF glucose levels. Intraseptal co-administration of glucose with morphine attenuated the memory impairment, and ECF glucose levels in the hippocampus decreased in a manner comparable to that seen in control rats. These data suggest that fluctuations in hippocampal ECF glucose levels may be a marker of mnemonic processing and support the view that decreases in extracellular glucose during memory testing reflect increased glucose demand during memory processing. PMID- 16343564 TI - A triterpenoid saponin possessing antileishmanial activity from the leaves of Careya arborea. AB - Bioguided-fractionation of the methanol extract of the leaves of Careya arborea led to isolation of a triterpenoid saponin, designated arborenin, and characterized as 3-O-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl-2 alpha,3beta-dihydroxy-taraxast-20-en-28-oic acid (1), together with desacylescin III (2). The structures were determined on the basis of extensive 2D NMR spectroscopic analysis. The saponin showed in vitro antileishmanial activity against Leishmania donovani (strain AG 83). PMID- 16343565 TI - Phytochemistry and antimycobacterial activity of Chlorophytum inornatum. AB - In a project to investigate plant derived natural products from the Liliaceae with activity against fast-growing strains of mycobacteria, we have identified two new metabolites from Chlorophytum inornatum. The active principle, a new homoisoflavanone (1) was identified as 3-(4'-methoxybenzyl)-7,8-methylenedioxy chroman-4-one. The metabolite assigned as 7-(1'-hydroxyethyl)-2-(2'' hydroxyethyl)-3,4-dihydrobenzopyran (2) was characterised by extensive 1- and 2D NMR spectroscopy. The antimycobacterial activity of this plant was mainly due to the homoisoflavonoid which exhibited minimum inhibitory values ranging from 16 256 microg/ml against four strains of fast-growing mycobacteria. PMID- 16343566 TI - Impediments to policy implementation: the offer of free installation of central heating to an elderly community has limited uptake. AB - OBJECTIVES: Increasing the warmth of homes in the UK to combat excess winter mortality has become an important part of public policy. This study examined the success of a local initiative to provide free installation of central heating for a group of elderly people. STUDY DESIGN: Cohort study. METHODS: The uptake of the offer of central heating was monitored through its different stages. Where possible, comments were invited from the elderly tenants to whom the offer was made; a sub-sample was interviewed about their views. RESULTS: Of 1181 tenants approached, only around one-quarter finally received central heating within the year. Reasons for the low uptake included inaccuracies in estimating those eligible as well as tenants' worries over upheaval and mess, health concerns and fear of increased heating costs. CONCLUSION: Successful policy implementation requires not only command of appropriate financial resources and the backing of local 'implementers', but also the local resources with which the policy will engage, especially local knowledge, and the 'targets' of the policy initiative, in this case the elderly tenants. PMID- 16343567 TI - Quality of training in public health in the UK: results of the first national training audit. AB - OBJECTIVES: Since 1997, UK public health training has undergone major changes, including the creation of specialist registrars (SpRs), introduction of a competency framework, admission of non-medical (specialist) trainees and National Health Service organizational change. It was therefore considered timely to audit the quality of this training. STUDY DESIGN: Cross-sectional survey of all UK public health trainees, conducted in April 2003. METHODS: The survey questions were based on 75 previously identified standards, with three sections: induction (30 standards), health protection (13 standards) and general training (32 standards). Results were calculated for the UK. Deaneries were compared on 10 key standards, as was the training of SpRs and specialist trainees. RESULTS: Two hundred and ninety responses were received (62% response rate). Only 16 (21%) of 75 standards were met by at least 80% of respondents, with problems in induction, health protection, secretarial facilities and examination support. Across 10 key standards (including initial welcome, trainer support, breadth/relevance of work and facilities), 59% indicated that their training had met at least eight standards. Results for individual deaneries were significantly different (P = 0.02), although 13 of 16 had median scores of eight out of 10, or over. Deaneries with specialists scored lower than those without (median scores eight vs nine, P = 0.003). Median specialist and SpR scores on the 10 key standards were seven and eight, respectively (P<0.001). In addition, SpRs were more likely to be 'on-call' [odds ratio (OR) = 66.8, 95% confidence interval (CI) 17.2-259.4, P<0.001] and to feel prepared for this role (OR = 10.7, 95% CI 1.4-79.8) than specialists. CONCLUSION: This was the first UK National Audit of Public Health Training. Few standards were achieved amongst respondents, although the levels set may be considered to be high and the response rate (62%) was less than optimal, potentially biasing results. Despite these caveats, recent organizational change in England appears to have led to significant training disruption among respondents. Nevertheless, repeating such an audit annually within deaneries could help to improve public health training throughout the UK. PMID- 16343568 TI - A stereological method for estimating the feto-maternal exchange surface area in the bovine placentome at 135 days gestation. AB - The feto-maternal relationships within the bovine placentome at 135 days gestation were investigated using unbiased stereology. This provides an estimate of the feto-maternal interface without physically separating these. The functional surface area, the volume of the fetal and maternal components, volumetric tissue shrinkage due to processing, and the feto-maternal surface amplification factor were estimated and quantified in standard units. In this healthy bovine pregnancy there were 105 placentomes with a total volume of 900 ml. The volumetric tissue shrinkage was 24% and the maternal: fetal tissue ratio overall was 0.92. The total feto-maternal exchange surface area was 18.5 m2 with a surface amplification factor of 108.1. Analysis of the coefficient of error from counting different numbers of points within the reference area suggested that 50% and 75% fewer points could be counted to achieve the result obtained by counting approximately 6000 points used in this study. PMID- 16343569 TI - Rapid molecular typing of Burkholderia pseudomallei, isolated in an outbreak of melioidosis in Singapore in 2004, based on variable-number tandem repeats. AB - An increase in the number of reported melioidosis cases was observed in the first 4 months of 2004. These cases were associated with a significant increase in case fatality rate compared with the past 5 years. In order to exclude the possibility of a single source, including the possibility of intentional release of Burkholderia pseudomallei, we applied a multiplex PCR-based multilocus variable number tandem repeat (VNTR) assay to determine the clonality of the clinical isolates. Our investigation indicated that a total of 30 different VNTR types could be distinguished in the 32 clinical isolates of B. pseudomallei obtained during this period, thus indicating that infection was unlikely to have occurred from a single source. Our experience underscores the usefulness of a rapid strain typing method in augmenting an epidemiological investigation into an infectious disease outbreak, particularly at a time where the intentional use of biological agents is a potential threat to public health. PMID- 16343570 TI - Intrathecal cytokine responses in Trypanosoma brucei rhodesiense sleeping sickness patients. AB - Intrathecal cytokine levels and blood-cerebrospinal fluid (CSF) barrier function were studied in 91 Trypanosoma brucei rhodesiense-infected patients. The CSF concentration of the cellular immune activation marker neopterin and the cytokines IL-6 and IL-10 were increased over control and post-treatment levels in all patients, with maximal levels observed in late-stage (meningoencephalitic) individuals. Analysis of CSF/serum concentration quotients indicated that IL-10 and neopterin were derived from central nervous system synthesis in at least 25% of the patients. Blood-CSF barrier dysfunction occurred in 64% of late-stage patients but not in early-stage patients. While the high level of neopterin observed in the late-stage patient CSF is indicative of widespread cellular activation, the increased levels of IL-6 and IL-10 suggest that counter inflammatory cellular responses may be important in the regulation of neuropathogenesis in late-stage human African trypanosomiasis. PMID- 16343571 TI - Characterization of Leishmania infantum strains in blood samples from infected dogs and humans by PCR-RFLP. AB - Characterization of Leishmania infantum is based on zymodeme analysis, which requires parasite isolation and therefore is not routinely employed. Moreover, the majority of strains in the Mediterranean Basin belong to zymodeme MON-1, and this is a major limitation for this technique in epidemiological studies in this region. We developed a PCR-RFLP method based on kDNA amplification, which was able to discriminate L. infantum strains directly from peripheral blood. Twenty eight samples were tested with this technique: four obtained from promastigote cultures, and 24 collected from dogs (18) and human donors (six) from traditionally endemic and newly endemic areas of northwestern Italy. Extracted DNAs were amplified using RV1-RV2 primers and PCR products were digested using two restriction enzymes separately: BsiY I and Mlun NI. Some patterns were specific to certain areas. In particular, the identity of PCR-RFLP patterns from a human patient from a newly endemic area and three dogs allow the confirmation of the autochthonous origin of this case. This approach could be applied to epidemiological studies in order to trace the diffusion of L. infantum within dog populations, as well as its transmission to humans. PMID- 16343572 TI - New initiatives against Africa's worms. AB - Since 1999, the funding available for the control of diseases of poverty (neglected diseases) has increased mainly due to leverage resulting from donations by the Bill and Melinda Gates Foundation and loans from the World Bank. Many countries have embarked on control programmes on a national scale due to drug donations by pharmaceutical companies through vertical programmes. The Schistosomiasis Control Initiative has expanded its operations to cover six countries in sub-Saharan Africa, but overlap of treatments between different vertical programmes is now a reality, and so care is needed to ensure that too many different drugs are not given together. Dialogue between programme managers has increased, and integration of some programmes may offer chances of synergy. PMID- 16343573 TI - Exact sample sizes needed to detect dependence in 2 x 3 tables. AB - Many medical and biological studies entail classifying a number of observations according to two factors, where one has two and the other three possible categories. This is the case of, for example, genetic association studies of complex traits with single-nucleotide polymorphisms (SNPs), where the a priori statistical planning, analysis, and interpretation of results are of critical importance. Here, we present methodology to determine the minimum sample size required to detect dependence in 2 x 3 tables based on Fisher's exact test, assuming that neither of the two margins is fixed and only the grand total N is known in advance. We provide the numerical tools necessary to determine these sample sizes for desired power, significance level, and effect size, where only the computational time can be a limitation for extreme parameter values. These programs can be accessed at . This solution of the sample size problem for an exact test will permit experimentalists to plan efficient sampling designs, determine the extent of statistical support for their hypotheses, and gain insight into the repeatability of their results. We apply this solution to the sample size problem to three empirical studies, and discuss the results with specified power and nominal significance levels. PMID- 16343574 TI - Multiallelic selection polymorphism. AB - The existence and stability of an internal (i.e., completely polymorphic) equilibrium for viability selection at a single multiallelic locus is investigated. Generations are discrete and nonoverlapping; the population is panmictic, monoecious, and diploid. Various necessary and sufficient conditions for the existence of an internal equilibrium are established and applied to the loss of alleles. Some necessary conditions for the existence of an asymptotically stable internal equilibrium are also established. All these conditions are simpler and yield general biological conclusions more easily than the classical necessary and sufficient conditions. PMID- 16343575 TI - Pyruvate reduces 4-aminophenol in vitro toxicity. AB - Pyruvate has been observed to reduce the nephrotoxicity of some agents by maintaining glutathione status and preventing lipid peroxidation. This study examined the mechanism for pyruvate protection of p-aminophenol (PAP) nephrotoxicity. Renal cortical slices from male Fischer 344 rats were incubated for 30-120 min with 0, 0.1, 0.25 or 0.5 mM PAP in oxygenated Krebs buffer containing 0 or 10 mM pyruvate or glucose (1.28 or 5.5 mM). LDH leakage was increased above control by 0.25 and 0.5 mM PAP beginning at 60 min and by 0.1 mM PAP at 120 min. Pyruvate prevented an increase in LDH leakage at 60- and 120-min exposure to 0.1 and 0.25 mM PAP. Pyruvate also prevented a decline in ATP levels. Glucose (1.28 and 5.5 mM) provided less protection than pyruvate from PAP toxicity. Total glutathione levels were diminished by 0.1 and 0.25 mM PAP within 60 and 30 min, respectively. Pyruvate prevented the decline in glutathione by 0.1 mM PAP at both time periods and at 30 min for 0.25 mM PAP. Pyruvate reduced the magnitude of glutathione depletion by 0.25 mM PAP following a 60-min incubation. Glutathione disulfide (GSSG) levels in renal slices were increased at 60 min by exposure to 0.25 mM PAP, while pyruvate prevented increased GSSG levels by PAP. Pyruvate also reduced the extent of 4-hydroxynonenal (4-HNE)-adducted proteins present after a 90-min incubation with PAP. These results indicate that pyruvate provided protection for PAP toxicity by providing an energy substrate and reducing oxidative stress. PMID- 16343576 TI - Nifedipine represses ion channels, transporters and Ca(2+)-binding proteins in hearts of spontaneously hypertensive rats. AB - The Ca(2+) antagonists nifedipine has been used for more than three decades to treat hypertension, but its effects on the transcriptional regulation of cardiac genes are basically unknown. We therefore studied expression of genes coding for ion channels, ion transporters and associated partners as well as Ca(2+)-binding proteins in ventricular tissue of normotensive and spontaneously hypertensive (SH) rats after repeated intraperitoneally (i.p.) dosing of nifedipine. Notably, we observed significant (P < 0.05) repression in transcript levels of most of the genes investigated, including cardiac Na(+), K(+), Ca(2+)-channels (L-type Ca(2+) channel, K(ir)3.4, K(ir)6.1, Na(v)1.5), ATP-driven ion exchangers (Na(+)-K(+) ATPase, NCX-1, PMCA 2 and 4, SERCA 2a and 2b) and their associated partners (phospholamban, RyR-2) as well as cytoskeletal proteins (alpha and beta-MHC, alpha cardiac and alpha skeletal actin, troponin T and I). Repression in transcript levels was, however, only seen in ventricular tissue of hypertensive animals. This points to fundamental differences in the mode of action of nifedipine in diseased and healthy animals. Indeed, this preponderance of repressed genes will promote disturbed ion homeostasis to result in contractile dysfunction. It is of considerable importance that repressed gene expression was also seen in end-stage human heart failure. We propose repression of cardiac specific gene expression as a hallmark of nifedipine treatment in hypertrophic hearts. PMID- 16343577 TI - TESS-based dose-response using pediatric clonidine exposures. AB - OBJECTIVE: The toxic and lethal doses of clonidine in children are unclear. This study was designed to determine whether data from the American Association of Poison Control Centers Toxic Exposure Surveillance System (TESS) could be utilized to determine a dose-response relationship for pediatric clonidine exposure. METHODS: 3,458 single-substance clonidine exposures in children <6 years of age reported to TESS from January 2000 through December 2003 were examined. Dose ingested, age, and medical outcome were available for 1550 cases. Respiratory arrest cases (n = 8) were classified as the most severe of the medical outcome categories (Arrest, Major, Moderate, Mild, and No effect). Exposures reported as a "taste or lick" (n = 51) were included as a dose of 1/10 of the dosage form involved. Dose ranged from 0.4 to 1980 (median 13) microg/kg. Weight was imputed based on a quadratic estimate of weight for age. Dose certainty was coded as exact (26% of cases) or not exact (74%). Medical outcome (response) was examined via logistic regression using SAS JMP (release 5.1). RESULTS: The logistic model describing medical outcome (P < 0.0001) included Log dose/kg (P = 0.0000) and Certainty (P = 0.045). CONCLUSION: TESS data can provide the basis for a statistically sound description of dose-response for pediatric clonidine poisoning exposures. PMID- 16343578 TI - The combined effect of spatial compounding and nonlinear filtering on the speckle reduction in ultrasound images. AB - Recently, a spatial compounding ultrasound imaging method was presented that utilizes a conventional 64-element phased array transducer with two unfocused pistons, each placed at one of the sides of the phased array transducer. This method is augmented here by inclusion of nonlinear filtering of the compounded images. The combined effects of the specific spatial compounding and nonlinear filtering on speckle reduction in the generated ultrasound images are studied and evaluated in two stages: First, the image quality is studied when nonlinear filtering is used as part of the spatial compounding. The study is performed by simulations using the Field II program, by processing several B-mode images of a kidney. The second stage compares the results obtained by the simulations to those obtained by in vitro laboratory experiments. Five different compounding strategies and two nonlinear filters, Gaussian and anisotropic diffusion, are investigated and evaluated in terms of image quality parameters-contrast and signal-to-noise ratio. It is shown that the combination of "averaging+nonlinear Gaussian filtering" produces the greatest improvement of image quality. When compared to a conventional phased array imaging system, the spatial compounding method that includes the conventional 64-element phased array transducer with two unfocused pistons, and employs the "averaging+nonlinear Gaussian filtering" strategy, obtains improvement in SNR that has reached 334%. Thus, though this method necessitates a somewhat wider probe, it produces significantly improved images. PMID- 16343579 TI - siRNAs against the Epstein Barr virus latency replication factor, EBNA1, inhibit its function and growth of EBV-dependent tumor cells. AB - The Epstein Barr virus (EBV) plays a role in maintenance of the tumor phenotype in a number of human cancers. The EBV latency replication factor, EBNA1, is required for persistence of the EBV episome, is anti-apoptotic, and is universally expressed in all EBV-associated tumors. Here, we show that EBNA1 specific siRNAs can inhibit EBNA1 expression and function. siRNAs were generated against three target sites in the EBNA1 messenger RNA, and two of these were found to inhibit EBNA1 expression from an ectopic EBNA1 expression cassette. EBNA1 siRNAs also inhibit endogenously expressed EBNA1 in EBV-positive epithelial and B-cell lines. Using a mini-EBV replication model, siRNA-mediated inhibition of EBNA1 expression suppressed the episomal maintenance function of EBNA1. Lastly, introduction of an EBNA1 siRNA into an EBV-positive tumor cell line inhibited tumor cell growth/survival. These data suggest that siRNAs against EBNA1 may have therapeutic value in EBV-associated diseases. PMID- 16343580 TI - Norovirus classification and proposed strain nomenclature. AB - Without a virus culture system, genetic analysis becomes the principal method to classify norovirus (NoV) strains. Currently, classification of NoV strains beneath the species level has been based on sequences from different regions of the viral genome. As a result, the phylogenetic insights of some virus were not appropriately interpreted, and no consensus has been reached to establish a uniform classification scheme. To provide a consistent and reliable scientific basis for classifying NoVs, we analyzed the amino acid sequences for the major capsid protein of 164 NoV strains by first using an alignment based on the predicted 3D structures. A Bayesian tree was generated, and the maximum likelihood pairwise distances of the aligned sequences were used to evaluate the results from the uncorrected pairwise distance method. Analyses of the pairwise distances demonstrated three clearly resolved peaks, suggesting that NoV strains beneath the species level can be classified at three levels: strain (S), cluster (C), and genogroup (G). The uncorrected pairwise distance ranges for S, C, and G were 0-14.1%, 14.3-43.8%, and 44.9-61.4%, respectively. A scheme with 29 genetic clusters [8 in genogroup 1 (G1), 17 in G2, 2 in G3, and 1 each in G4 and G5] was defined on the basis of the tree topology with the standards provided and was supported by the distance analysis. Of these, five clusters in G2 and one in G1 are newly described. This analysis can serve as the basis for a standardized nomenclature to genetically describe NoV strains. PMID- 16343581 TI - Solid-liquid separation of faecal sludge using drying beds in Ghana: implications for nutrient recycling in urban agriculture. AB - This study investigated the possibility of recycling nutrients in human excreta and municipal solid waste for use in agriculture. It reports on the use of drying beds in separating solid and liquid fractions of faecal sludge (FS) so that the solids can be co-composted and the organic matter and part of the nutrients captured for urban agriculture. Sludge influent onto drying beds, percolate effluent, and dewatered sludge (biosolids) were monitored over eight loading cycles in 2002. The unplanted drying beds were made of 15 cm of sand (0.2-0.6mm diameter) and 25 cm gravel (10 and 19 mm diameter). The loading rate of sludge ranged from 196 to 321 kg total solids (TS) /m(2)y. Biosolids with TS 20% were obtained after an average drying time of 2 weeks. The drying beds retained 80% of solids and 100% of helminth eggs. The biosolids had average organic matter content of 61%; hence, allowing for co-composting with biodegradable organic solid waste for hygienisation. The process is being investigated further to attain higher efficiency and reliability. PMID- 16343582 TI - Optimum design and operation of primary sludge fermentation schemes for volatile fatty acids production. AB - This paper presents a model-knowledge based algorithm for optimising the primary sludge fermentation process design and operation. This is a recently used method to obtain the volatile fatty acids (VFA), needed to improve biological nutrient removal processes, directly from the raw wastewater. The proposed algorithm consists in a heuristic reasoning algorithm based on the expert knowledge of the process. Only effluent VFA and the sludge blanket height (SBH) have to be set as design criteria, and the optimisation algorithm obtains the minimum return sludge and waste sludge flow rates which fulfil those design criteria. A pilot plant fed with municipal raw wastewater was operated in order to obtain experimental results supporting the developed algorithm groundwork. The experimental results indicate that when SBH was increased, higher solids retention time was obtained in the settler and VFA production increased. Higher recirculation flow-rates resulted in higher VFA production too. Finally, the developed algorithm has been tested by simulating different design conditions with very good results. It has been able to find the optimal operation conditions in all cases on which preset design conditions could be achieved. Furthermore, this is a general algorithm that can be applied to any fermentation-elutriation scheme with or without fermentation reactor. PMID- 16343583 TI - Optimization of phosphate removal in anodizing aluminium wastewater. AB - The wastewater produced after brightening and anodizing aluminium has high concentrations of phosphates and sulphates. The addition of MgO in a first physico-chemical wastewater treatment step makes the selective recovery of phosphates in the form of magnesium phosphates feasible, which may be reused as fertilizer. The proposed wastewater treatment process allows manufacturers to reduce more than 70% of the volume of the precipitate in the sedimentation reactor and more than 50% of the weight of the final disposal sludge. In this study, the use of an alternative low-grade MgO (LG-MgO) as a source of magnesium, which is cheaper than pure MgO, is investigated. The phosphate concentration and pH of the treated wastewater is controlled by the formation and precipitation of newberyite or bobierrite as a function of the magnesium source added. According to experimental data, a reaction mechanism is proposed. Although LG-MgO reacts more slowly than pure MgO and it is necessary to add 3-4 times the stoichiometric amount, this procedure has considerable economic and technical advantages. PMID- 16343584 TI - Formation, adsorption and separation of high molecular weight disinfection byproducts resulting from chlorination of aquatic humic substances. AB - A significant portion of the unidentified disinfection byproducts (DBPs) in chlorinated drinking water can be attributed to high molecular weight (MW) chlorinated DBPs (above 500 Da) that may have adverse health effects. In this work, issues on the formation, adsorption and separation of high MW-chlorinated DBPs were investigated by introducing radioactive (36)Cl into humic substance samples. The results show that the amount of high MW-chlorinated DBPs during chlorination decreased with the increase of contact time from 1 to 120 h, increased with the increase of pH from 5.5 to 9.5, and was less in the ultrafiltered samples from Suwannee River fulvic acid than from Suwannee River humic acid. The high MW-chlorinated DBPs were found to be effectively adsorbed by activated carbon and be possibly reduced to Cl(-) by activated carbon, but not to be readily desorbed from the activated carbon. Those high MW-chlorinated DBPs were demonstrated to be incapable of resolution into discrete peaks by any of the three liquid chromatography columns studied. The significant implications of these results are discussed. PMID- 16343585 TI - Design optimization of a self-cleaning moving-bed bioreactor for seawater denitrification. AB - The hydrodynamic behavior and denitrification capacity of a down flow self cleaning moving bed bioreactor (MBBR) using floating packed-bed carriers were investigated. Water mixing in the MBBR was determined by tracer studies, which showed a completely stirred tank reactor. Eleven different types of patented plastic packed-bed carriers were selected based on their density, shape and specific surface area. Video studies of the carriers were used to determine operating conditions and select the Cascade 1A and Tri-Pack 1 carriers for denitrification experiments. Both carriers showed a similar performance level in terms of the denitrification capacity of the resulting MBBR. The improvement in liquid circulation is thought to explain the high denitrification rates that reached 27 gNm(-2)d(-1) and the maintenance of a thin biofilm. Increasing the overall liquid velocity profile led to an increase of up to 30% in the denitrification rate in conditions with a 1-month-old biofilm. This MBBR design seemed to control biofilm development and could easily be scaled up to denitrify seawater or freshwater systems. PMID- 16343586 TI - Simulation of the operation of detention tanks. AB - The performance of detention tanks with different characteristics (volume, on line and off-line arrangement) has been evaluated according to the results of a continuous simulation. The conceptual simplified model for sewer system simulation (COSMOSS) model has been used to simulate the operation of the tanks. The differences between the performance of on-line and off-line tanks and the influence of the characteristics of different catchments have been examined. According to the results of the simulations detention tanks demonstrated good performances in total suspended solids retention and this evenience can certainly help to prevent water pollution of receiving water bodies in urban areas, even if the differences between the catchments, especially regard to the first flush effect, influence the performance of the tanks. Anyway considerable good efficiencies can be obtained with tank volumes of about 30-50 m(3)/ha(imp), in terms of number, maximum concentrations and duration of overflows, generally not guaranteed only with overflow devices. PMID- 16343587 TI - Simultaneous removal of perchlorate and nitrate from drinking water using the ion exchange membrane bioreactor concept. AB - This work evaluates the feasibility of the ion exchange membrane bioreactor (IEMB) concept for the simultaneous removal of perchlorate and nitrate from drinking water, when nitrate is present in the ppm range and perchlorate in the ppb range. The IEMB concept combines Donnan dialysis and simultaneous biological degradation of both pollutants. Membrane transport studies showed that Donnan dialysis is suitable for obtaining water with concentrations of perchlorate and nitrate below the recommended levels. However, the pollutants were accumulated in a receiving stream, thus requiring additional treatment before disposal. On the other hand, the IEMB process operated with hydraulic retention times ranging from 1.4 to 8.3h in the water compartment, proved to remove effectively perchlorate and nitrate while preserving the water composition with respect to other ions, thus avoiding secondary contamination of the treated water. For a polluted water stream containing 100 ppb of ClO(4)(-) and 60 ppm of NO(3)(-), the concentrations of both ions in the treated stream were kept below the recommended levels of 4 ppb for ClO(4)(-) and 25 ppm for NO(3)(-). The IEMB system was operated under ethanol limitation, but even under these conditions, an increase of the perchlorate and nitrate concentrations in the treated water was not observed for up to 6 days. PMID- 16343588 TI - Wind, rain and bacteria: The effect of weather on the microbial composition of roof-harvested rainwater. AB - The microbiological and chemical quality of tank-stored rainwater is impacted directly by roof catchment and subsequent run-off contamination, via direct depositions by birds and small mammals, decay of accumulated organic debris, and atmospheric deposition of airborne micro-organisms and chemical pollutants. Previous literature reports on roof water quality have given little consideration to the relative significance of airborne micro-organisms. This study involved analyses of direct roof run-off at an urban housing development in Newcastle, on the east coast of Australia. A total of 77 samples were collected during 11 separate rainfall events, and microbial counts and mean concentrations of several ionic contaminants were matched to climatic data corresponding to each of the monitored events. Conditions both antecedent to, and those prevailing during each event, were examined to investigate the influence of certain meteorological parameters on the bacterial composition of the roof water and indirectly assess the relative contribution of airborne micro-organisms to the total bacterial load. Results indicated that airborne micro-organisms represented a significant contribution to the bacterial load of roof water at this site, and that the overall contaminant load was influenced by wind velocities, while the profile (composition) of the load varied with wind direction. The implications of these findings to the issues of tank water quality and health risk analysis, appropriate usage and system design are discussed. PMID- 16343589 TI - Metal toxicity in municipal wastewater activated sludge investigated by multivariate analysis and in situ hybridization. AB - The toxicity of three heavy metals (copper, zinc and nickel) on an activated sludge biomass was studied by measuring several parameters of microbiological activity and the dynamics of microbial community composition. The effects on the microflora in the activated-sludge tank were evaluated by dosing the tank with different metal concentrations and a non-inhibiting concentration of copper for a 2-week period. The relationship between biological effects and chemical measurements was elucidated by principal component analysis (PCA). The biochemical data suggested a higher sensitivity of nitrifiers to these metals than heterotrophs. This was confirmed by fluorescent in situ hybridization (FISH) analysis on the microbial community. The impact of copper was displayed by biomass deflocculating phenomena (pin-point) and a decrease in the beta-proteo bacteria component. In addition, the zone settling velocity (ZSV) was shown not to be a good indicator of chronic toxicity phenomena. Finally, the sequence of biomass metal accumulation capability was assessed, and corresponds to Cu>Ni>Zn. PMID- 16343590 TI - A different approach for predicting H(2)S((g)) emission rates in gravity sewers. AB - All detrimental phenomena (mal odors, metal corrosion, concrete disintegration, health hazard) associated with hydrogen sulfide in gravity sewers depend on the rate of H(2)S emission from the aqueous phase to the gas phase of the pipe. In this paper a different approach for predicting H(2)S((g)) emission rates from gravity sewers is presented, using concepts adapted from mixing theory. The mean velocity gradient (G=gamma SV/micro; S is the slope, V the mean velocity), representing mixing conditions in gravity flow, was used to quantify the rate of H(2)S((g)) emission in part-full gravity sewers. Based on this approach an emission equation was developed. The equation was verified and calibrated by performing 20 experiments in a 27-m gravity-flow experimental-sewer (D=0.16 m) at various hydraulic conditions. Results indicate a clear dependency of the sulfide stripping-rate on G(1) (R(2)=0.94) with the following overall emission equation: where S(T) is the total sulfide concentration in the aqueous phase, mg/L; w the flow surface width, m; A(cs) the cross-sectional area, m(2); T the temperature, degrees C; K(H) the Henry's constant, molL(-1)atm(-1); and P(pH2S) the partial pressure of H(2)S((g)) in the sewer atmosphere, atm. PMID- 16343591 TI - Reaction of acrolein with 2'-deoxyadenosine and 9-ethyladenine--formation of cyclic adducts. AB - Treatment of 2'-deoxyadenosine with acrolein at pH 4.6 in 37 degrees C affords unstable adducts containing either one or two fused ring systems where the hydroxypropano units are derived from acrolein. Since the use of 2' deoxyadenosine resulted in the creation of at least four diastereoisomers for the adduct made up of two fused rings, therefore, for identification and assignment of the products, 9-ethyladenine was used instead as the starting material in the reaction. The products, 3E and 4E, were structurally characterised by UV, mass spectrometry and NMR spectroscopy. PMID- 16343592 TI - Quantitative structure-property relationships on photodegradation of polybrominated diphenyl ethers. AB - By partial least squares (PLS) regression, quantitative structure-property relationship (QSPR) models were developed for photodegradation rates (k(p)) and quantum yields (Phi) of polybrominated diphenyl ethers (PBDEs) in methanol/water (8:2), and photodegradation rates in pure methanol by UV light in the sunlight region, respectively. Quantum chemical descriptors computed by PM3 Hamiltonian were used as predictor variables. The cross-validated Q(cum)(2) values for three optimal QSPR models of PBDEs are above 0.90 (remarkably higher 0.50), indicating good predictive abilities for logk(p) and logPhi values of PBDEs. The QSPR results show that logk(p) values of PBDEs in methanol/water (8:2) and in pure methanol are governed by different molecular structural descriptors, respectively, which implies that photodegradation rates of PBDEs are affected by the characteristics of solution in which it takes place. PMID- 16343593 TI - Toenail cerium levels and risk of a first acute myocardial infarction: the EURAMIC and heavy metals study. AB - The association between cerium status and risk of first acute myocardial infarction (AMI) was examined in a case-control study in 10 centres from Europe and Israel. Cerium in toenails was assessed by neutron activation analysis in 684 cases and 724 controls aged 70 years or younger. Mean concentrations of cerium were 186 and 173 microg/kg in cases and controls, respectively. Cerium was positively associated with low socio-economic status, smoking, mercury, zinc and scandium (p0.001). Cases had significantly higher levels of cerium than controls after adjustment for age and centre (case-control ratio 1.074; 95% CI 1.002 1.151) and increased in further adjustment for other cardiovascular risk factors 1.085; 95% CI 1.025-1.149. The risk after adjustment for age and centre was higher with increasing cerium levels (p for trend=0.02). After adjustment for BMI, history of hypertension, smoking, alcohol intake, diabetes, family history of CHD, beta-carotene, lycopene, alpha-tocopherol, selenium, mercury and scandium, the OR for the highest quintile was 1.43 (95% CI 0.85-2.41; p-trend 0.08). When we applied this same model in non-smokers the odds ratios in the 4th and 5th quintiles of cerium as compared with the lowest were 2.09 (95% CI 1.05 4.16) and 2.81 (95% CI 1.21-6.52), respectively, p-trend 0.011. Our results suggest that toenail cerium levels may be associated with an increased risk of AMI, but more research is warranted to shed further light and fully understand the plausibility and public health implications of these findings. PMID- 16343594 TI - Comparison of biodegradation of poly(ethylene glycol)s and poly(propylene glycol)s. AB - The biodegradation of poly(ethylene glycol)s (PEGs) and poly(propylene glycol)s (PPGs), both being major by-products of non-ionic surfactants biodegradation, was studied under the conditions of the River Water Die-Away Test. PEGs were isolated from a water matrix using solid-phase extraction with graphitized carbon black sorbent, then derivatized with phenyl isocyanate and determined by HPLC with UV detection. PPGs were isolated from a water matrix by liquid-liquid extraction with chloroform, then derivatized with naphthyl isocyanate and determined by HPLC with fluorescence detection. The primary biodegradation of both PEGs and PPGs reached approximately 99% during the test. The tests show different biodegradation pathways of PEG and PPG. During PEG biodegradation, their chains are shortened leading to the formation of ethylene glycol and diethylene glycol. During PPG biodegradation, no short-chained biodegradation products were found. PMID- 16343595 TI - Toxicity on crustaceans and endocrine disrupting activity on Saccharomyces cerevisiae of eight alkylphenols. AB - In the last few years many concerns have been raised regarding the environmental safety of alkylphenol polyethoxylate surfactants (APnEOs). They are widely used in detergents, paints, herbicides and many other formulated products. It has been estimated that 60% of APnEOs end up in the aquatic environment; they are biodegradable and transformed into alkylphenols, such as nonylphenol and octylphenol that are hydrophobic and tend to accumulate. In the present study, acute and chronic aquatic toxicity and the estrogenic activity of the following eight alkylphenols were assessed: 4-nonylphenol, 4-octylphenol, 4-nonylphenol-10 ethoxylate, 4-tert-octylphenol, POE (1 to 2)-nonylphenol, POE (6)-nonylphenol, POE (3)-tert-octylphenol and POE (9 to 10)-tert-octylphenol. The toxic potential was measured on the crustaceans Daphnia magna and Ceriodaphnia dubia, while the estrogenic activity was determined by using the YES-test with the strain Saccharomyces cerevisiae RMY326. The results showed that the exposure of crustaceans to the eight xenoestrogens investigated caused both acute and chronic effects. The EC50 values found for C. dubia at 48 h were compared to D. magna at 24h and, gave a first indication about the toxic activity of the compounds investigated, that is better expressed in the long-term. In fact, chronic data showed a strong increase in toxicity with EC50 values one or two orders of magnitude lower than the acute values. The results of the YES-test showed that nonylphenol, octylphenol and 4-tert-octylphenol were the most estrogenic and the bioassay was able to detect their estrogenicity at very low concentrations (ng microg/l). PMID- 16343596 TI - Preliminary data on polybrominated diphenyl ethers (PBDEs) in farmed fish tissues (Salmo salar) and fish feed in Southern Chile. AB - Polybrominated diphenyl ethers (PBDEs) have become an issue of global concern. Recent studies have shown that farmed salmon can accumulate high levels of brominated compounds in their tissues and consequently there is a growing concern on its industrial and public health impacts. Little information is found in the international literature on PBDEs in the biotic compartment of the Southern Hemisphere. This paper reports the levels of several PBDE congeners found in the tissues of farmed fish from five different farming areas of Southern Chile. PBDEs were analyzed by HRGC-MS. More analytical data were obtained by analyzing these same pollutants in fish feed. Our results indicate a general trend of PBDE levels averaging 1.46 ng g(-1) wet weight (wwt). The observed congeneric distribution that resulted was quite similar to data previously reported in the open literature. PBDE profiles were found to be dominated by BDE 47. No correlation was observed between levels found in the tissues and the lipid content in such tissues, although a high correlation with the fish feed data was observed indicating that this could probably be the main PDBE entry source into fish, although other sources cannot be excluded. Even though the samples were obtained from different geographical areas, they presented fairly similar profiles, indicating a potential common source. We concluded that PBDE levels in the farmed Chilean salmon are in the low average range of values published in the open literature. PMID- 16343597 TI - Pleistocene human footprints from the Willandra Lakes, Southeastern Australia. AB - Human and other hominid fossil footprints provide rare but important insights into anatomy and behavior. Here we report recently discovered fossil trackways of human footprints from the Willandra Lakes region of western New South Wales, Australia. Optically dated to between 19-23 ka and consisting of at least 124 prints, the trackways form the largest collection of Pleistocene human footprints in the world. The prints were made by adults, adolescents, and children traversing the moist surface of an ephemeral soak. This site offers a unique glimpse of humans living in the arid inland of Australia at the height of the last glacial period. PMID- 16343598 TI - Serum from aged F344 rats conditions the activation of young macrophages. AB - There is considerable controversy about the molecular mechanisms responsible for the variations in innate immunity associated with age. While in vivo, aged animals and humans react to an inflammatory signal with an excessive production of pro-inflammatory cytokines, studies in vitro generally show that this response is attenuated in macrophages from old individuals. In an effort to examine possible extrinsic factors that might affect the response of macrophages to lipopolysaccharide (LPS), we have challenged peritoneal macrophages obtained from young rats with sera obtained from rats of different ages. Our results indicate that the serum from aged rats significantly impairs the capacity of young macrophages to induce tumor necrosis factor-alpha (TNF-alpha) production, while at the same time it increases the basal levels of interleukin-6 (IL-6). The effect of serum from aged donors on TNF-alpha secretion requires pre-incubation and is sensitive to heat inactivation. In contrast, the stimulating effect on IL 6 is resistant to heat, and thus should not be due to a protein factor. Therefore, our results indicate that the age-related changes in macrophage activity are not only the consequence of intrinsic changes, but there also appears to be a modulatory effect imparted by the external milieu. PMID- 16343599 TI - An assay on the effect of preliminary restoration tasks applied to a large TENORM wastes disposal in the south-west of Spain. AB - A large industrial wastes disposal site, where two phosphate rock processing plants release their wastes, located close to Huelva town (SW of Spain), has been partially submitted to restoration as a preliminary step in a possible decomissioning process. Due to the high natural radioactivity contents of these wastes, this repository is considered as a radiological anomaly, being actually considered as TENORM (technically enhanced naturally occurring radioactive materials). The efficiency of this restoration from the radiological point of view according to the new European regulatory framework was evaluated in this work. The results allow to conclude that, as a consequence of the partial restoration works, the external dose rate has been drastically reduced above the repository system. Nevertheless, special attention must be paid on the occupational factor to be applied to workers on the unrestored system. The application of a dosimetric model allows the prediction of the negative effects of using certain industrial wastes as a cover system in this restoration/mitigation task. PMID- 16343600 TI - Characteristic of polycyclic aromatic hydrocarbon concentrations and source identification for fine and coarse particulates at Taichung Harbor near Taiwan Strait during 2004-2005. AB - Fine (PM(2.5)) and coarse (PM(2.5-10)) particulate concentrations of ambient air polycyclic aromatic hydrocarbons (PAHs) were measured simultaneously from February 2004 to January 2005 at the Taichung Harbor (TH) sampling site near Taiwan of central Taiwan. Particle-bound polycyclic aromatic hydrocarbons (PAHs) were collected on quartz filters; the collected sample was Soxhlet extracted with a dichloromethane (DCM)/n-hexane mixture (50/50, v/v) for 24 h, and then the extracts were analyzed by gas chromatography-mass spectrometry (GC-MS). The results indicated that vehicle emissions, coal combustion, incomplete combustion and pyrolysis of fuel and oil burning were the main source of PAHs near Taiwan Strait of central Taiwan. Diagnostic ratios and principal component analysis (PCA) were also used to characterize and identify PAHs emission sources in this study. PMID- 16343601 TI - Mercury content in vegetation and soils of the Almaden mining area (Spain). AB - The Almaden area is an exceptional case of mercury concentration in the world, where cinnabar is the predominant mineralised form. Mining activities have been carried out over the past two thousand years and once this activity is concluded, a restoration of the environment is necessary for the affected areas. That implies the selection, optimisation and application of the most appropriate techniques focused on the recovery of contaminated areas. Phytotechnology is one of the proposed techniques, which considers the use of plants due to the low environmental impact and their proved efficiency. In this framework, it is necessary to study the behaviour of mercury in the soil-plant system and to determine the mercury absorption capacity of the different Almaden plant species. Therefore, field and laboratory studies have been carried out to determine the easily available forms of mercury in ten test plots in the Almaden area and the resulting data is related to the mercury accumulation in different plant species of the studied territory. Special attention is given to the easily available forms of mercury in the analysed soils and used as the transfer factor for the Hg availability index that relates the amount of mercury in a plant with the total amount of mercury in a soil and with the easily available forms. The result of this experimental work is selecting the plant species suitable to be used in a future implementation of phytotechnologies in this area. PMID- 16343602 TI - Strategies to lower methyl mercury concentrations in hydroelectric reservoirs and lakes: A review. AB - Mercury (Hg) concentrations in fish in lakes are elevated due to increased global cycling of Hg. A special case of elevated Hg concentrations in fish occurs in new hydroelectric reservoirs because of increased rates of converting Hg in the environment into methyl mercury (MeHg). People and wildlife that eat fish from hydroelectric reservoirs have an elevated risk of accumulating too much MeHg. Demand for electrical energy is leading to the creation of new reservoirs. In 2005, Canada derived 60% of its electricity from hydroelectric reservoirs. As a result, hydroelectric companies and governing agencies are exploring strategies to lower MeHg contamination. Strategies may involve lowering the source of Hg before flooding, the rate of Hg methylation, or MeHg bioaccumulation and biomagnification. Possible strategies reviewed in this article include selecting a site to minimize impacts, intensive fishing, adding selenium, adding lime to acidic systems, burning before flooding, removing standing trees, adding phosphorus, demethylating MeHg by ultraviolet light, capping and dredging bottom sediment, aerating anoxic bottom sediment and waters, and water level management. A preventative strategy is to limit the flooded area, especially wetland areas. Flooded upland areas that contain less carbon produce MeHg for a shorter time than wetland areas. Run-of-the-river reservoirs contain lower MeHg concentrations than reservoirs that flood vast areas, at the cost of exporting MeHg downstream. Managing water levels to flush systems during times of peak MeHg production may have benefits for the reservoir, but also transports MeHg downstream. Intensive fishing can lower MeHg in food webs by increasing fish growth rate. Additions of selenium can lower MeHg bioaccumulation, but the mechanisms are not well established and excess selenium causes toxicity. Liming can lower fish Hg concentrations in lakes acidified with sulphuric and nitric acid. Burning before flooding can lower the production of MeHg, but MeHg bioaccumulation may increase. The most promising strategy will be one that is agreeable to all affected people. PMID- 16343603 TI - Platelet activation predicts recurrent ischemic events after percutaneous coronary angioplasty: a 6 months prospective study. AB - INTRODUCTION: An increasing amount of evidence indicates that platelet reactivity, despite a standard anti-thrombotic therapy, is a potential risk factor for recurrent myocardial ischemia in patients with coronary artery disease. We now hypothesize that this condition, measured by collagen-epinephrine (CEPI) or collagen-ADP (CADP) closure times (CT) by Platelet Function Analyzer (PFA-100), may predict the recurrence of coronary events after percutaneous coronary intervention (PCI). MATERIALS AND METHODS: CEPI and CADP-CT were measured 30+/-8 h after PCI in 175 consecutive patients admitted with a diagnosis of stable angina (n=94) or acute coronary syndromes (n=81) and prospectively followed up for a mean period of 6 months. We stratified the patients in accordance to both the CEPI-CT ( 190 s), reflecting the intensity of cycloxygenase inhibition by aspirin and the distribution into quartiles for CADP CT. RESULTS: CEPI-CT<190 s as well as CADP-CT<82 s were associated with a higher rate of clinical recurrence (hazard ratio 8.5, p<0.001 and 22.9, p<0.001, respectively). Multivariate analysis after adjustment for other risk factors confirmed that the lowest CADP-CT quartile significantly correlates with the risk of recurrent coronary events (hazard ratio 36.5, p<0.01), as well as CEPI-CT<190 s (hazard ratio 6.7, p=0.01). CONCLUSIONS: An enhanced platelet function after PCI when measured under high shear rates by PFA-100 is an independent predictor of a worst clinical outcome, even during a short term follow-up and may help in patients risk stratification. PMID- 16343604 TI - Surgical morbidity associated with radical trachelectomy and radical hysterectomy. AB - OBJECTIVES: To examine intra-operative, short and longer term morbidity associated with Radical Trachelectomy (RT) within our cohort of patients, compared with conventional Radical Hysterectomy (RH). METHODS: A retrospective case note review comprising 29 RT and 50 RH patients. Patients who required adjuvant treatment were excluded. Operative data, short-term and long-term complications were recorded. Long-term problems were included only if they had been of persistent nature. RESULTS: The median age and follow up period for the RH group were 40 years and 60 months and for RT group 30.5 years and 25 months. There were significant differences between RH and RT groups in median operative time (260 versus 187 min), blood loss (1000 versus 400 ml), transfusion requirement (75% versus 12%), analgesic requirement (8 versus 3.5 days) and hospital stay (11 versus 6 days). Bladder hypotonia requiring prolonged catheterisation was more frequent in RH group (P = 0.004). There was no apparent difference in psychosexual problems between the two groups (17 versus 16%). Complications of deep dyspareunia (P = 0.009), excessive vaginal discharge (P = 0.01), and upper thigh parasthesia (P = 0.05) were noted to be significantly higher in the RT group. Specific problems encountered in the RT group included; dysmenorrhea (24%), irregular menstruation (17%), recurrent candidiasis (14%), cervical suture problems (14%), isthmic stenosis (10%) and prolonged amenorrhea (7%). CONCLUSION: RT is associated with less operative and short-term morbidity compared with conventional RH. In addition to specific complications associated with RT, in our experience, there are long-term morbidities that are not as commonly observed in RH. PMID- 16343605 TI - Worsening effect of partial sleep deprivation on indomethacin-induced gastric mucosal damage. AB - The present study was to investigate the roles of cyclooxygenase-1 and -2 (COX-1 and COX-2) and prostaglandin (PG) on gastric mucosal integrity of partially sleep deprived (PSD) rats. A slowly moving drum was used to induce PSD. The PG levels in the gastric mucosa of PSD rats, with or without indomethacin or rofecoxib treatment, were determined. Exogenous prostaglandin E (PGE) analog, misoprostol, was administered to PSD rats to investigate the modulating effect of PG in indomethacin-induced gastric damage. It was observed that COX-1 mRNA and protein were up-regulated in the gastric mucosa of PSD rats. Selective COX-2 inhibition by rofecoxib failed to decrease mucosal PGE2 levels nor to affect mucosal integrity in both PSD and sleep undisturbed rats. However, indomethacin, a COX-1 preferential non-selective COX inhibitor, significantly reduced mucosal PGE2 content and produced more severe mucosal damage in PSD rats than in the controls. The deleterious effect of indomethacin on gastric mucosal integrity of PSD rats was significantly attenuated with the administration of misoprostol. These results suggest that PSD enhances COX-1 biosynthesis of gastroprotective PGE2 as an adaptive response of the stomach to stress. The administration of non selective COX inhibitors to subjects with chronic sleep deprivation may induce more gastric damages. PMID- 16343606 TI - Neural localization of semantic context effects in electromagnetic and hemodynamic studies. AB - Measures of electrical brain activity (event-related potentials, ERPs) have been useful in understanding language processing for several decades. Extant data suggest that the amplitude of the N400 component of the ERP is a general index of the ease or difficulty of retrieving stored conceptual knowledge associated with a word, which is dependent on both the stored representation itself, and the retrieval cues provided by the preceding context. Recordings from patients with brain damage, intracranial recordings, and magnetoencephalographic data implicate a (probably large portion of) the left temporal lobe as the largest source of the N400 semantic context effect, with a substantial but lesser contribution from the right temporal lobe. Event-related functional magnetic resonance (fMRI) studies using semantic context manipulations are dominated by observations of greater hemodynamic activity for incongruent sentence completions or semantically unrelated words than congruent or related words, consistent with the direction of the ERP effect. The locations of the hemodynamic effects show some variability across studies, but one commonly identified region is the left superior temporal gyrus, which is compatible with the electrophysiological results. A second commonly identified region in the fMRI studies is the left inferior frontal gyrus, which does not appear to make a substantial contribution to the N400 effect. PMID- 16343607 TI - Novel method for titanium crown casting using a combination of wax patterns fabricated by a CAD/CAM system and a non-expanded investment. AB - OBJECTIVES: For titanium casting, most commercial investments for titanium recommend casting at a low mold temperature to reduce oxidation. However, the thermal expansion values of the molds at low casting temperatures may be insufficient. The purpose of the current study was to investigate the possibility of obtaining accurate titanium crown casts using wax pattern fabricated by a CAD/CAM system with a non-expanded mold. METHODS: Three types of experimental magnesia-based investments (A, B and C) were made and their properties were evaluated for dental use. Two kinds of wax patterns for full-coverage coping crowns (S-0: cement space of 0 microm; S-20: cement space of 20 microm) were fabricated using a commercial CAD/CAM system. A traditional method (TM) using inlay wax was performed for comparison. The investment for titanium casting was decided from the fundamental data of experimental investments. Titanium crowns were replaced on the stone die and the thickness of the cement layer was evaluated. RESULTS: There were no significant differences for the setting time and setting expansion among the experimental investments, but the aluminous cement content played a role in hardening and contracting the mold. The fit of the titanium crowns differed significantly between the TM and the CAD/CAM system. The ranges of thickness obtained from the TM, S-0 and S-20 were 20.78-357.88 microm, 25.12-107.46 microm and 17.84-58.92 microm, respectively. SIGNIFICANCE: High quality titanium crown casting was obtained using a combination of wax patterns fabricated by a CAD/CAM system and a non-expanded MgO-based investment. PMID- 16343608 TI - Does sediment grain size affect diatom grazing by harpacticoid copepods? AB - Estuarine soft sediments support a diverse group of eukaryotic and prokaryotic organisms though the role of the sediment per se for the functioning of these organisms remains largely unknown. The present study aimed to test the effect of sediment grain size on the grazing activities of harpacticoid copepods. In controlled experiments, two common intertidal harpacticoid species (Paramphiascella fulvofasciata and Nitokra spinipes) were each offered a mix of two benthic diatom species (Navicula phyllepta and Seminavis robusta) in different sedimentary conditions. Several microcosms were created using a variety of sediment types, including fine silt (<63 microm), coarser grained sands (125 250, 250-450, 100-300 microm), artificial 'sediments' of glass beads (250-500, 2000 microm) and even the absence of sediment was tested. The diatoms were enriched in the stable carbon (13)C to facilitate tracing in the harpacticoids. Both copepod species were able to graze on the diatoms with highest uptake when sediment was absent. In contrast, both harpacticoid species showed no uptake in silty conditions. In general, grazing was favoured when mean sediment grain size increased. The strong negative effect of fine grains on the grazer's efficiency can be explained by the resulting differences in the structure (and accessibility) of the diatom biofilm on the one hand and the mobility of the grazer on the other hand. In view of the subtle equilibrium between primary producers and grazers, these results might have important implications for the effect of siltation of tidal flats due to, e.g., human activities. PMID- 16343609 TI - Environmental prognostics: an integrated model supporting lysosomal stress responses as predictive biomarkers of animal health status. AB - The potential prognostic use of lysosomal reactions to environmental pollutants is explored in relation to predicting animal health in marine mussels, based on diagnostic biomarker data. Cellular lysosomes are already known to accumulate many metals and organic xenobiotics and the lysosomal accumulation of the carcinogenic polycyclic aromatic hydrocarbon 3-methylcholanthrene (3-MC) is demonstrated here in the hepatopancreatic digestive cells and ovarian oocytes of the blue mussel. Lysosomal membrane integrity or stability appears to be a generic indicator of cellular well-being in eukaryotes; and in bivalve molluscs it is correlated with total oxygen and nitrogen radical scavenging capacity (TOSC), protein synthesis, scope for growth and larval viability; and inversely correlated with DNA damage (micronuclei), as well as lysosomal swelling (volume density), lipidosis and lipofuscinosis, which are all characteristic of failed or incomplete autophagy. Integration of multiple biomarker data is achieved using multivariate statistics and then mapped onto "health status space" by using lysosomal membrane stability as a measure of cellular well-being. This is viewed as a crucial step towards the derivation of explanatory frameworks for prediction of pollutant impact on animal health; and has facilitated the development of a conceptual mechanistic model linking lysosomal damage and autophagic dysfunction with injury to cells, tissues and the whole animal. This model has also complemented the creation and use of a cell-based bioenergetic computational model of molluscan hepatopancreatic cells that simulates lysosomal and cellular reactions to pollutants. More speculatively, the use of coupled empirical measurements of biomarker reactions and modelling is proposed as a practical approach to the development of an operational toolbox for predicting the health of the environment. PMID- 16343610 TI - An arabinogalactan isolated from the stems of Cistanche deserticola induces the proliferation of cultured lymphocytes. AB - A purified polysaccharide ACDP-2 was isolated from water extract of the stems of Cistanche deserticola. Chemical and spectroscopic analyses indicated that ACDP-2 is a highly branched arabinogalactan polymer that composes of linked d galactopyranose and d-glucopyranose, which contains predominantly a branching point at the 6-carbon. The branched side-chains compose of terminal-, 1,5-, and 1,3,5-linked arabinofuranosyl residues. ACDP-2 showed an effect in stimulating the immune response, which when applied onto the cultured mouse lymphocytes induced the cell proliferation in a dose-dependent manner. PMID- 16343611 TI - Demineralized bone matrix gelatin as scaffold for osteochondral tissue engineering. AB - To develop a single-unit osteochondral tissue with demineralized bone matrix gelatin (BMG), rabbit chondrocytes were cultured on demineralized bone matrix gelatin for 6 weeks. The engineered osteochondral tissue was characterized with histology, immunolocalization, TEM, SEM, biochemical assay, and gene expression analysis. About 1.3mm viable neo-cartilage was produced on demineralized BMG. RT PCR, immunohistochemistry, TEM, biochemical assay, and histology revealed hyaline like cartilage with zonal layers, intense type II collagen expression, and abundant proteoglycan content formed upon BMG compared with normal cartilage. But hydroxyproline content and type I collagen gene and protein expressions were significantly lower. We consider engineering cartilage tissue with chondrocytes cultured on allogenic demineralized BMG is a good approach for osteochondral tissue engineering. PMID- 16343612 TI - Polypyrrole doped with 2 peptide sequences from laminin. AB - In the field of neural tissue engineering, electrically conducting, biocompatible surfaces are of great interest. Over the past several decades conducting polymers have been studied as candidate surfaces because they fit these criteria. Several attempts have been made to combine the conductivity and biocompatibility of conducting polymers with biomolecules that could promote specific cell attachment and growth. In this report the laminin fragments CDPGYIGSR (p31) and RNIAEIIKDI (p20) are used as dopants in electropolymerization of the conducting polymer polypyrrole (PPy). The electrical properties of the resulting films are analyzed by impedance spectroscopy and cyclic voltammetry and compared to gold. PPy/p20 surfaces consistently demonstrate the lowest impedance and largest charge capacity for a given deposition charge. Next, in vitro studies using primary neurons cultured in a defined media and primary astrocytes in a serum containing media were performed; neuron density and neurite length, as well as astrocyte density, were quantified. Surfaces doped with a combination of the two peptides (PPy/p20-p31) consistently supported the highest neuronal density. It is shown that surfaces doped with the laminin fragment p20 had significantly longer primary neurites than either the p31 doped or poly(styrenesulfonate) doped PPy surfaces. Finally, the astrocyte studies demonstrate that PPy surfaces have significantly less astrocyte adhesion in culture than the common electrode material, gold. PMID- 16343613 TI - Photoimmobilized array of panel cells for assay of antibodies. AB - Antibodies in blood are checked with panel blood cells before blood transfusion. In this investigation, for the first time, a panel cell-microarray was prepared by using a photoimmobilization method. Different types of red blood cells were microarrayed on a plate. A water-soluble photoreactive polymer as a matrix was synthesized by the coupling reaction of azidoaniline with poly(2 methacryloyloxyethylphosphorylcholine-co-methacrylic acid). The polymer was mixed with cells and the mixtures were microspotted on substrate and photoirradiated after drying in air. For the antibody assay, monoclonal antibodies or human serum was added to the cell-arrayed plate and adsorbed antibodies were detected by horseradish peroxidase-labeled secondary antibody, which recognized the adsorbed antibodies. Antibodies specifically adsorbed on the immobilized cells as expected. The aggregation method has been available for this type of assay, but extensive experience was needed to apply it correctly. The method using a cell array will be useful for antibody detection. PMID- 16343614 TI - Biological characterisation of vascular grafts cultured in a bioreactor. AB - In this study, the development is described of a tissue-engineered construct mimicking the structure of a natural blood vessel. Smooth muscle cells (SMC) were cultured under pulsatile flow conditions in porous tubular scaffolds composed of crosslinked type I insoluble collagen and insoluble elastin. Under these dynamic culture conditions, average wall shear rate, systolic and diastolic pressures and pressure wave-forms comparable to conditions in the human carotid artery were obtained. Culturing of SMC in tubular scaffolds under dynamic conditions resulted in enhanced tissue formation compared to static conditions. Higher SMC numbers, a more homogeneous distribution of SMC throughout the scaffolds and higher collagen mRNA expression levels were found when cells were cultured under dynamic compared to static conditions. mRNA expression levels of markers of proliferation and apoptosis showed that the higher cell numbers in the scaffolds cultured under dynamic conditions can be explained by increased cell proliferation but not by decreased apoptosis. Glucose consumption and lactate formation by the cells showed that cell metabolism was more aerobic under dynamic compared to static conditions. Lining of the dynamically cultured constructs with a luminal monolayer of endothelial cells might result in vessels suitable for in vivo applications. PMID- 16343615 TI - Differential expression of the receptor tyrosine kinase EphB4 and its ligand Ephrin-B2 during human placental development. AB - Normal placentation involves the development of an utero-placental circulation following the migration of the extravillous cytotrophoblasts into the decidua and invasion of the spiral arteries, which are thereby transformed into large vessels of low resistance. Given the documented role of the receptor tyrosine kinase EphB4 and its ligand ephrin-B2 in the establishment of the embryonal vascular network, we hypothesized that these molecules are also instrumental in the development of the human placenta. Monitoring the expression during placental development revealed that in first trimester and term placentae both molecules are equally expressed at the RNA level. In contrast, the protein levels were significantly reduced during gestation. Immunohistochemistry revealed a distinct localization of the EphB4 and ephrin-B2 proteins. EphB4 was predominantly expressed in the villous syncytial trophoblast layer and in a subset of intravillous capillaries. Prominent expression was also observed in the extravillous cytotrophoblast giant cells. In contrast, ephrin-B2 expression was detected in the villous cytotrophoblast and syncytial trophoblast cell layers, as well as initially in all intravillous capillaries. Strong expression was also observed in extravillous anchoring cytotrophoblast cells. Hypoxia is a major inducer of placental development. In vitro studies employing trophoblast-derived cell lines revealed that predominantly ephrin-B2 expression is induced by hypoxia, however, in an Hif-1alpha independent manner. These experiments suggest that EphB4 and ephrin-B2 are instrumental in the establishment of a functional placental structure and of the utero-placental circulation. PMID- 16343616 TI - Recent advances in Ca(2+)-dependent Ras regulation and cell proliferation. AB - Our understanding of the mechanisms whereby growth factors stimulate cell proliferation through the Ras pathway stems largely from studies of the canonical pathway involving recruitment of Ras activators and inhibitors to the vicinity of receptor tyrosine kinases via phosphotyrosine-binding adaptor proteins. Ca(2+) has seldom joined the party, despite the identification of phospholipase Cgamma and Ca(2+) entry as receptor tyrosine kinase-dependent signals. Mechanisms by which Ca(2+) can directly influence Ras activity have remained relatively elusive. Similarly, the mechanisms whereby Ca(2+) modulates the cell cycle have been equally murky, and yet there are some interesting parallels in the role of Ras and Ca(2+) in cell cycle re-entry. This review focuses on a number of novel mechanisms that link Ca(2+) with the regulation of Ras activity and signaling output. Their collective discovery adds to the complexities of Ras regulation and raises further questions about the role of Ca(2+) signals in Ras-dependent cell proliferation. PMID- 16343617 TI - Changes in tracheo-bronchial sensory neuropeptide receptor gene expression pattern in rats with cisplatin-induced sensory neuropathy. AB - An attenuated neurogenic broncho-constriction underpinned by a decrease in sensory neuropeptide release has been shown to be characteristic of cisplatin induced neuropathy. The present work was to explore if beyond neuropeptide release, cisplatin at a treatment schedule attaining sensory neuropathy, produced changes in the expression of the receptors of sensory neuropeptides such as somatostatin, calcitonin gene-related peptide (CGRP) and substance P (SP) in bronchial tissue of the rat. Twenty-four Wistar rats were divided into three groups. The animals in the "Treatment groups 1 and 2" were given cisplatin (1.5mgkg(-1)) and mannitol (75mgkg(-1)) over 5 days. The rats in the "Control" group were given mannitol+isotonic saline. Four animals from each group were used to study the expression pattern of the neuropeptide receptors in bronchial tissue. The levels of somatostatin receptor 4 (SSTR 4), neurokinin 1 (NK1), neurokinin 2 (NK2) and CGRP receptor expression were examined by quantitative real time polymerase chain reaction (RT-PCR) method, 11 and 22 days after the last cisplatin/vehicle dose. The cisplatin treatment significantly increased plasma somatostatin immunoreactivity and the expression of SSTR4 receptor detected both on the 11th and 22nd post-treatment days with no change in either CGRP, NK1, and NK2 receptor gene expression or plasma CGRP and substance P levels. We conclude that cisplatin neuropathy is accompanied by an increase in plasma somatostatin immunoreactivity with an increase in SSTR4 expression in rats. PMID- 16343618 TI - Identification of allelic polymorphism in the caprine IGHA gene. AB - Variation in the immunoglobulin heavy alpha chain (IGHA) constant region has been reported in a number of species. In this study, the IGHA gene was investigated in goats using PCR-single-strand conformational polymorphism (SSCP) analysis and DNA sequencing. Three novel sequences were identified from 111 Boer and Angora goats. Either one or two sequences were detected in individual goats, and all the sequences shared high homology to the published ovine and bovine IGHA sequences. These sequences were predicted to encode three amino acid sequences, two with a longer hinge region and one with a shorter hinge region. The variation reported here may affect the structure of the hinge and hence the function of IgA. PMID- 16343619 TI - Identification of linear plasmid pAM1 in the flavonoid degrading strain Actinoplanes missouriensis(T) (DSM 43046). AB - By pulsed-field gel electrophoresis, a linear DNA element of about 100 kb was identified in Actinoplanes missouriensis(T) DSM 43046, which grows on the flavonoids hesperidin, rutin and quercetin, and which contains a CO forming quercetinase. Among six Actinoplanes species and strains tested, including A. globisporus(T) DSM 43857, A. philippinensis(T) DSM 43019, A. brasiliensis(T) DSM 43805, A. auranticolor(T) DSM 43031, and A. utahensis(T) DSM 43147, only the A. missouriensis strain exhibited such a genetic element. The linear plasmid, named pAM1, has proteins covalently attached to its 5'-ends like other linear replicons of actinomycetes. Attempts to cure pAM1 failed, however a mutant with reduced plasmid content was obtained, which showed reduced ability to degrade the flavonoid rutinosides rutin and hesperidin. Plasmid pAM1 is the first extrachromosomal genetic element identified in an Actinoplanes species and may be useful to develop genetic tools for biotechnologically important Actinoplanes strains. PMID- 16343620 TI - Reproductive success of passerines exposed to polychlorinated biphenyls through the terrestrial food web of the Kalamazoo River. AB - The eastern bluebird (Sialia sialis) and the house wren (Troglodytes aedon) were identified as ecological receptors of concern due to exposure and potential effects stemming from polychlorinated biphenyl (PCB) contamination in floodplain soils of the Kalamazoo River Superfund Site, Michigan, USA. Measures of population health were compared at a contaminated and a less-contaminated reference location. During this 3-year study, productivity of bluebirds was significantly less at the downstream location than at the reference location. Hatching success, clutch size, and predicted brood size were significantly less in early clutches of house wrens at the more contaminated location than at the upstream reference location, but fledging success was significantly greater at the contaminated location. Studies concurrent to the study presented here reported that concentrations of PCBs in the tissues and diets of the passerine birds were less than the predicted threshold for adverse effects. The results of our study, taken along with the measured exposure data, suggest that other factors in addition to PCB exposure such as habitat, prey availability, small sample size, and cocontaminants were likely causes of the differences that were observed at the two locations. PMID- 16343621 TI - Correlation between 96-h mortality and 24-h acetylcholinesterase inhibition in three grass shrimp larval life stages. AB - Three life stages of larval grass shrimp were tested to determine whether acetylcholinesterase (AChE) activity expressed as 24-h sublethal effect endpoints (EC20 and EC50) could be used to predict 96-h mortality (lowest observable effect concentration (LOEC) and LC50) for shrimp exposed to three organophosphate insecticides. With regard to mortality, newly hatched larvae and 18-day-old larvae were the most sensitive in the malathion and azinphosmethyl exposures. In the chlorpyrifos exposures, newly hatched larvae and postlarvae were the most sensitive life stages. Results of the 24-h AChE inhibition tests showed that newly hatched larvae were generally more sensitive in the three organophosphate exposures. A regression analysis of the EC50's and LC50's yielded the strongest correlation with R2=0.987 (correlation coefficient=0.994 and 95% confidence intervals 0.969-0.999). The LOEC/EC20 relationship yielded R2=0.962. For these grass shrimp life stages and pesticides, sublethal effect endpoints could be used as a predictor of 96-h mortality. PMID- 16343622 TI - Rearrangement of only one human IGHV gene is sufficient to generate a wide repertoire of antigen specific antibody responses in transgenic mice. AB - In recent years, mice carrying human IG transgenes are being generated for the production of human monoclonal antibodies as an alternative approach to the conventional use of mouse or chimeric-humanized antibodies. Theoretically, the size of the repertoire of human antibodies that these mice could produce would be critically dependent on the number of human V genes introduced in the transgene. This could be the case for BABkappa and BABkappa,lambda transgenic mice, which carry several genes from the human IGK (BABkappa), and IGK and IGL (BABkappa,lambda) loci, but only five human IGHV genes and the entire IGHD-IGHJ cluster linked to two human IGHC (IGHM-IGHD) genes. We analyzed the expressed human IG genes in 30 IgM-secreting hybridomas generated from transgenic mice immunized either with soluble proteins (human IgM coupled to KLH) or with cells (human PBMC, tumour cell lines or rat cells transfected with human CD69). The results show that all hybridoma cells analyzed rearranged exclusively the IGHV1-2 gene, in contrast with naive spleen B cells that used three out of the five IGHV genes present in the transgene. The configuration of the rearranged CDR3 region revealed a much higher heterogeneity in the heavy chains. A variety of IGHJ and IGHD genes were used in hybridomas, and somatic mutations were also seen in some hybrids. Regarding the rearranged light chains genes, it was a much higher variety in the use of V and J genes in both, kappa and lambda chains, than in the heavy chain, and also in the level of mutation. The results indicate that only one IGHV gene is sufficient to generate a wide repertoire of antigen specific antibody responses. Thus, efforts aimed at the generation of new transgenic mice should focus more on the integrity of the D/J region and on the DNA regions regulating somatic hypermutation, rather than on the number of V genes present in the transgene. PMID- 16343623 TI - A novel designed single domain antibody on 3-D structure of ricin A chain remarkably blocked ricin-induced cytotoxicity. AB - Ricin A chain (RA), an N-glycosidase, is able to fatally disrupt protein synthesis by attacking the Achilles heel of the ribosome RNA (rRNA). As specific immunotoxins, emergence of inhibitors for RA may obtain access to antagonistics against ricin intoxication and contribute to ameliorate the concomitant side effects. Many experimental results showed that the engineered VHs, which possessed solubility, stability, small size and consequently easier to express, purify and manipulate in vitro, were self and long-lived molecules compared to synthetic peptides. In this study, based on the crystal structure of RA, a novel recombinant human single-domain antibody expressing a polypeptide against RA in the CDR3 loop (named rVH(PT)) was obtained using computer-guided molecular design method. Theoretically, rVH(PT) could penetrate deeply into the active cleft of RA and act as a potent antagonist analogue to block the RA-rRNA interaction. Followed results showed that the recombinant VH(PT) was easily expressed of high yield production and in a partially soluble fusion form in Escherichia coli. In vitro cytotoxicity experiments demonstrated that it possessed remarkable ability to block ricin-induced cytotoxicity. This study highlights the potential of human VH to display biostructure and biofunction of peptides designed on RA functional domain and could be useful in developing new antidotes with potential therapeutic uses to neutralize unintended exposure to ricin. PMID- 16343624 TI - Localized orbital amyloidosis involving the lacrimal sac and nasolacrimal duct. AB - PURPOSE: To report the case of a 70-year-old man who presented with tearing in his left eye and a firm palpable lump in the area overlying his left lacrimal sac. DESIGN: Retrospective interventional case report. METHODS: Noninvasive diagnostic evaluation followed by external dacryocystorhinostomy, histopathologic studies, and systemic evaluation. RESULTS: The patient was found to have idiopathic localized amyloidosis limited to the lacrimal sac and nasolacrimal duct. CONCLUSION: The localized form of amyloidosis is rare, typically involves the head and neck without systemic manifestations, and carries an excellent prognosis. Previous reports of orbital amyloidosis have described involvement of the lacrimal gland, extraocular muscles, and the cranial nerves. To our knowledge, this is the first report of a patient with nasolacrimal duct obstruction secondary to amyloid deposition in the lacrimal sac and fossa. PMID- 16343625 TI - Idiopathic juxtafoveal retinal telangiectasis: new findings by ultrahigh resolution optical coherence tomography. AB - OBJECTIVE: To investigate the capabilities of ultrahigh-resolution optical coherence tomography (UHR OCT); to compare with the commercially available OCT standard-resolution system, StratusOCT, for imaging of idiopathic juxtafoveal retinal telangiectasis (IJT); and to demonstrate that UHR OCT provides additional information on disease morphology, pathogenesis, and management. DESIGN: Retrospective, observational, interventional case series. PARTICIPANTS: Nineteen eyes of 10 patients diagnosed with IJT in at least one eye. METHOD: All patients were imaged with UHR OCT and StratusOCT at the same visit. A subset of patients was also imaged before and after treatment of IJT. MAIN OUTCOME MEASURES: Ultrahigh- and standard-resolution cross-sectional tomograms of IJT pathology. RESULTS: Using both standard- and ultrahigh-resolution OCT, we identified the following features of IJT: (1) a lack of correlation between retinal thickening on OCT and leakage on fluorescein angiography, (2) loss and disruption of the photoreceptor layer, (3) cystlike structures in the foveola and within internal retinal layers such as the inner nuclear or ganglion cell layers, (4) a unique internal limiting membrane draping across the foveola related to an underlying loss of tissue, (5) intraretinal neovascularization near the fovea, and (6) central intraretinal deposits and plaques. In 63% of cases, the presence of abnormal vessels and a discontinuity of the photoreceptor layer correlated with visual acuity. CONCLUSIONS: Ultrahigh-resolution OCT improves visualization of the retinal pathology associated with IJT and allows identification of new features associated with it. Some of these features, such as discontinuity of the photoreceptor layer, are revealed only by UHR OCT. PMID- 16343626 TI - Field-dependent aniseikonia associated with an epiretinal membrane a case study. AB - PURPOSE: Aniseikonia is a binocular anomaly in which the two eyes perceive images of different sizes and/or shapes. It is usually assumed to be constant as a function of visual field angle (VFA) (i.e., angular distance from the line of sight). This is correct for optically induced aniseikonia, such as the aniseikonia that is associated with anisometropia and probably also pseudophakia. The purpose of this article is to show that if the aniseikonia is of retinal origin, then the aniseikonia may no longer be constant as a function of VFA (i.e., field-dependent aniseikonia). DESIGN: Case report, with the patient having a unilateral epiretinal membrane. METHODS: The aniseikonia was measured in vertical and horizontal directions with a customized version of the Aniseikonia Inspector software. The VFA was made variable by changing the dimensions of the comparison targets in the direct comparison procedure. MAIN OUTCOME MEASURE: Aniseikonia as a function of VFA. RESULTS: The patient exhibited good repeatable aniseikonia, ranging from 23% to 2.5% for VFAs ranging from 0.36 degrees to 5.7 degrees . Higher angles had lower aniseikonia. A control subject did not show this field-dependent aniseikonia. CONCLUSIONS: Aniseikonia may vary with VFA due to a retinal cause such as an epiretinal membrane. The problem with field dependent aniseikonia is that it cannot be corrected fully with conventional optics, which exhibit an approximately constant magnification as a function of VFA. Nevertheless, by correcting 5% to 10% aniseikonia, which showed up in the VFA measurement range at 2 degrees to 3 degrees , our patient had improved visual comfort, especially for reading. PMID- 16343627 TI - Changes in retinal neovascularization after pegaptanib (Macugen) therapy in diabetic individuals. AB - OBJECTIVE: To study effects of intravitreal pegaptanib (Macugen) on retinal neovascularization. DESIGN: Retrospective analysis of a randomized clinical trial. PARTICIPANTS, INTERVENTION, AND MAIN OUTCOME MEASURES: Individuals with retinal neovascularization identified from a multicenter, randomized, controlled trial evaluating pegaptanib for treatment of diabetic macular edema, with a best corrected visual acuity letter score between 68 and 25 (approximate Snellen equivalent between 20/50 and 20/320) and receiving a sham injection or intravitreal pegaptanib (0.3 mg, 1 mg, 3 mg) administered at study entry, week 6, and week 12, with additional injections and/or focal photocoagulation as needed during the ensuing 18 weeks, up to a maximum of 6 pegaptanib/sham therapies, were evaluated. Scatter panretinal photocoagulation before study enrollment was permitted, but not within 6 months of randomization and study entry. Changes in retinal neovascularization were assessed on fundus photographs and fluorescein angiograms graded at a reading center in a masked fashion. RESULTS: Of 172 participants, 19 had retinal neovascularization in the study eye at baseline. Excluding 1 who had scatter photocoagulation 13 days before randomization and 2 with no follow-up photographs, 1 of the remaining 16 subjects had panretinal photocoagulation during study follow-up. Of these 16 subjects, 8 of 13 (62%) in a pegaptanib treatment group (including the one receiving panretinal photocoagulation), 0 of 3 in the sham group, and 0 of 4 fellow (nonstudy) eyes showed either regression of neovascularization on fundus photographs or regression or absence of fluorescein leakage from neovascularization (or both) at 36 weeks. In 3 of 8 with regression, neovascularization progressed at week 52 after cessation of pegaptanib at week 30. CONCLUSIONS: Most subjects with retinal neovascularization at baseline assigned to pegaptanib showed regression of neovascularization by week 36. These findings suggest a direct effect of pegaptanib upon retinal neovascularization in patients with diabetes mellitus. PMID- 16343628 TI - A useful augmented lateral tarsal strip tarsorrhaphy for paralytic ectropion. AB - OBJECTIVE: Patients with paralytic ectropion and lagophthalmos may experience keratitis and may pose a functional and aesthetic surgical challenge. Various methods are used to reduce the vertical palpebral aperture, including lateral tarsal strip (LTS) or a lateral tarsorrhaphy. We modified the LTS to differentially shorten and elevate the lower lid more than the upper: an augmented LTS tarsorrhaphy (aug-LTS-T). This study aimed to evaluate the technique. DESIGN: Prospective noncomparative surgical trial in which preoperative and postoperative symptoms, margin reflex distances, vertical palpebral aperture (PA), lagophthalmos, and corneal findings were recorded. The data were analyzed at 6 months after surgery using the Wilcoxon sign-rank test for nonparametric data. PARTICIPANTS: Fourteen consecutive adult patients (15 eyelids) with chronic lagophthalmos and paralytic ectropion. METHODS: Patients underwent aug-LTS-T. This consisted of a long strip (10-15 mm) that is attached to the outer temporal orbital rim, at a point higher than a conventional LTS. It included removal of a small part of the upper eyelid anterior lamella laterally to pass the long strip up high enough. MAIN OUTCOME MEASURES: Improvement of symptoms, reduction of lower margin reflex distance, lagophthalmos, and improvement of corneal signs. RESULTS: Minimum follow-up was 6 months. There was a significant reduction in PA (P = 0.005) and lagophthalmos (P = 0.0002) with improvement of corneal signs (14 of 15 eyelids = 93%). Surgery was successful anatomically in 14 of 15 eyelids (93%) with low morbidity. CONCLUSIONS: We describe the augmented LTS tarsorrhaphy and find it effective in the treatment of severe lower eyelid ectropion resulting from facial palsy. PMID- 16343629 TI - Manganese exposure: neuropsychological and neurological symptoms and effects in welders. AB - Manganese exposure reportedly may have an adverse effect on CNS function and mood. Sixty-two welders with clinical histories of exposure to manganese were compared to 46 matched regional controls chosen at random from a telephone directory. The following tests were given: Wechsler Adult Intelligence Scale (WAIS-III), Wechsler Memory Scale (WMS-III), Boston Naming, WRAT-3, Cancellation H, Trail Making Tests A and B, Auditory Consonant Trigrams, Stroop, Rey Osterreith, Animal Naming, Controlled Oral Word Association (COWAT), Test of Memory Malingering, Rey 15-item, Fingertapping, Grooved Pegboard, Dynamometer, Visual Attention Test, Lanthony d-15 Color Vision, Vistech Contrast Sensitivity, and Schirmer strips. The controls were administered a shorter battery of tests and the Rey-Osterreith, Animal Naming and some of the subtests of the WAIS-III, WMS-III were not administered. Mood tests, given to both groups, included the Symptom Checklist-40, Symptom Checklist-90-R, Profile of Mood Scale, Beck Depression Inventory II, and Beck Anxiety Inventory. Forty-seven welders and 42 controls were retained for statistical analysis after appropriate exclusions. Results showed a high rate of symptom prevalence and pronounced deficits in motor skills, visuomotor tracking speed and information processing, working memory, verbal skills (COWAT), delayed memory, and visuospatial skills. Neurological examinations compared to neuropsychological test results suggest that neuropsychologists obtain significantly more mood symptoms overall. Odds ratios indicate highly elevated risk for neuropsychological and neurological symptomatology of manganism. Mood disturbances including anxiety, depression, confusion, and impaired vision showed very high odds ratios. Neurological exams and neuropsychological tests exhibit complementarity and differences, though neuropsychological methods may be more sensitive in detecting early signs of manganism. The present study corroborates the findings of our previous study in another group of welders. PMID- 16343630 TI - The metal- and DNA-binding activities of Helicobacter pylori NikR. AB - The pathogenic bacteria Helicobacter pylori require nickel as a cofactor of the enzymes urease and hydrogenase. One of the proteins that controls nickel homeostasis in this organism is Helicobacter pylori NikR (HpNikR), a homologue of nickel-dependent transcription factors from other organisms, which regulates the expression of multiple proteins such as the urease structural subunits and itself. To examine the properties of this protein, metal analysis was used to demonstrate that HpNikR can bind stoichiometric nickel or copper, and electronic absorption spectroscopy revealed that HpNikR binds nickel with picomolar affinity in what is likely a conserved square-planar site. In vitro DNA-binding assays revealed that HpNikR can bind directly to the promoter region of the ureA operon in response to nickel, and the location of the binding site was defined. Nickel also induces DNA binding to the nikR promoter sequence but the complex is much weaker. These experiments suggest that HpNikR directly controls the expression of multiple genes by binding to separate DNA sequences, and the possible mechanisms for differential regulation are discussed. PMID- 16343631 TI - DFT and the electromerism in complexes of iron with diatomic ligands. AB - A reliable procedure is proposed for assigning the electronic structures for large biologically-relevant systems, where the size of the model confines one to the use of density functional theory (DFT) methods, and where the risk of over interpreting DFT-derived molecular orbitals and spin densities still exists. The proposed approach focuses on the use of the only DFT-derived parameter that is unanimously recognized to be reliable: the geometry. We examine DFT-derived O-O bond lengths in formally ferrous-dioxygen models, and compare them to bond lengths in free, non metal-bound, dioxygen, superoxide and peroxide moieties. Likewise, we compare the N-O bond lengths within ferrous-nitrosyl {FeNO}7 models, with the same parameter in free NO+, NO*, and HNO species. This allows a calibrated, straightforward way of assigning the electronic structure in systems where electromerism makes detailed single-reference molecular orbital analysis unreliable. PMID- 16343632 TI - Synthesis and characterization of new palladium-clotrimazole and palladium chloroquine complexes showing cytotoxicity for tumor cell lines in vitro. AB - New palladium complexes of chloroquine (CQ) and clotrimazole (CTZ) have been prepared, characterized, and evaluated against four tumor cell lines in vitro. [Pd (CQ)2Cl2] (1) was synthesized by the reaction of PdCl2(CH3CN)2 with CQ, and the [Pd (CTZ)2Cl2] (2) complex by a similar reaction. The new compounds were characterized by a combination of FAB-MS (fast atom bombardment-mass spectrum), elemental analysis, molar conductivity, IR, and NMR spectroscopy. The solid-state structure of 2 has been determined by X-ray crystallography. 2 crystallizes in the monoclinic space group P(2(1)/c), with a = 21.100(4) A, b = 13.408(3) A, c = 22.642(5) A. The structure refinement converged at R1 = 0.0728, wR2 = 0.1918. The cytotoxicity of these two complexes for the tumor cell lines, PANC-1, SKBR-3, MDA MB231 and HT-29, was compared with that of the original ligands. Ligation of palladium to CTZ led to an increase in the IC50, although a three-fold reduction in the IC50 of CQ was observed on ligation to the metal when tested against the MDA-MB231 cell line. PMID- 16343633 TI - Microbiology of public telephones. AB - OBJECTIVES: There is an increasing interest in antibacterial products intended for applications, such as in carpets, hand lotions, wallpaper adhesives, gloves and fabrics used in the home and hospital environments. A range of materials impregnated with various types of antimicrobial agents, such as triclosan and silver-zeolite additive is also being evaluated for use in plastics, air filters, toys, tooth-brushes, pagers and telephones. However, there is not enough microbiological data to suggest the need for sterilization of telephones and similar hand-contact objects. METHODS: Bacteriological sampling of 50 public telephones was performed from May to November. RESULTS: Twelve different types of bacteria were found on the surface of telephones. The level of bacterial contamination for the telephone mouthpiece was increased to its highest point in October from its lowest value in August. It was also found that the microbial contamination of mouthpiece was about twice the contamination of earpiece. CONCLUSIONS: These results showed that public telephones are contaminated with various types of microorganisms and the level of contamination changes significantly with time. PMID- 16343634 TI - Validation of a Pseudomonas aeruginosa porcine model of septic shock. AB - OBJECTIVES: To develop a standardized bacteraemic porcine model of septic shock with cardiovascular and immunological profiles similar to those observed in human clinical states. METHODS: Sepsis was induced by an intravenous challenge of 18 anaesthetized pigs with live Pseudomonas aeruginosa. The pulmonary arterial pressure was monitored and the bacterial infusion was stopped when the systolic pulmonary arterial pressure reached 45 mmHg. Septic shock was treated with fluid resuscitation and epinephrine infusion. The haemodynamic parameters and the rate of different inflammatory cytokines were recorded during 6 h of observation. RESULTS: The mean+/-SD cardiac output increased from 2.4+/-1.2 to 5.7+/-2.1 L/min while the mean+/-SD systemic vascular resistance index decreased from 1957+/-744 to 709+/-221 dyn/s/cm5/m2. The pharmacokinetic profile of the inflammatory cytokines was similar to the one observed in human studies. CONCLUSIONS: The control of the systolic pulmonary arterial pressure during a P. aeruginosa infusion leads to a hyperdynamic, reproducible cardiovascular profile similar to the one observed in human septic shock. Since the immunological profile of the inflammatory cytokines is also similar to the human one, this standardized porcine model appears to be appropriate for experimental research concerning sepsis. PMID- 16343635 TI - TLR4 polymorphism in Iranian patients with brucellosis. AB - OBJECTIVE: Brucellosis is a zoonosis of both public health and economic significance in most developing countries. Polymorphisms in Toll-like receptor-4 (TLR4) have been reported to be associated with a blunted immune response to microbial pathogens. Information regarding any association between genetic variation of TLR4 and susceptibility to brucellosis is not available in the literatures. The main purpose of this research is to evaluate the role of polymorphic alleles of TLR4 gene in susceptibility to brucellosis. MATERIALS AND METHODS: In this case-control study, 198 patients with brucellosis and 111 healthy volunteers matched for sex, age and geographic area were evaluated by genotyping for polymorphism in TLR4 gene (Asp299Gly) using amplification refractory mutation system (ARMS)-PCR method. RESULTS: Allele 896G was more prevalent in patients with brucellosis compared to healthy controls (33.6% vs. 20.7%, P=0.000003). Also the frequency of G allele of TLR4 gene was significantly higher in male patients with brucellosis compared to the same sex in control group (36% vs. 21.7%, P=0.00005). Multiple logistic regression analysis demonstrated that male patients heterozygous at allele G gene had a significantly higher risk for brucellosis with an odds ratio of OR 2.89, 95% CI: 1.79-4.69, P<0.0001). CONCLUSION: This study is the first to show an association between genetic polymorphism in TLR4 gene and susceptibility to brucellosis. PMID- 16343636 TI - Positive mental health-related impacts of the SARS epidemic on the general public in Hong Kong and their associations with other negative impacts. AB - OBJECTIVES: To investigate 2 positive impacts (better family/social support and lifestyle changes) and their associations with other negative impacts. STUDY DESIGN AND PARTICIPANTS: A telephone survey using random telephone numbers and a structured questionnaire was conducted in Hong Kong general population. Eight hundred and eighteen Hong Kong residents of age 18-60 years completed the interview (response rate: 65%) at around end of June 2003, the early ending phase of the epidemic. RESULTS: Over 60% of the respondents stated that they cared more about the family members' feelings; about 30%-40% stated that they found their friends, their family members more supportive or having more sharing with others when not feeling happy; only a few percent felt the opposite. Further, about 2/3 of the respondents paid more attention to their mental health. About 35%-40% took more time to rest, for relaxation or doing exercise. These items were, in general, negatively associated with post-traumatic stress, perception of stress and other perceptions that were related to SARS. CONCLUSIONS: SARS have brought some positive impacts on social/family support, mental health awareness and lifestyle changes. These positive impacts were associated with other relevant negative impacts and might be important cushions of the negative impact. PMID- 16343637 TI - Epidemiology, risk factors and outcome of nosocomial infections in a Respiratory Intensive Care Unit in North India. AB - OBJECTIVE: To determine the epidemiology, risk factors and outcome of infections in a Respiratory Intensive Care Unit (RICU) of a tertiary care institute in northern India. METHODOLOGY: Prospective, observational clinical study. RESULTS: The study included 201 patients (1285 patient days) admitted to RICU over a period of one-and-a-half years. A total of 77 infections were identified in 67 patients (33.5%). The infections included pneumonia (23%), sepsis of unknown origin (10.5%), bacteremia (7.5%), urinary tract infections (1.5%), catheter related blood stream infections (1%) and Clostridium difficile colitis (1%). The most commonly identified organisms were the Acinetobacter species (34.8%), Pseudomonas aeruginosa (23.9%) and Escherichia coli (15.2%). The median length of stay in patients with and without infection was 13 days (interquartile range, IQR, 28) and 4 days (interquartile range, IQR, 3), respectively (p<0.0001). Multivariate analysis showed the following risk factors for ICU-acquired infection: the admitting diagnosis of infection (odds ratio [OR] 3.3; 95% confidence intervals [CI] 1.06-10.1), length of stay in the RICU (OR, 1.2; 95% confidence intervals [CI] 1.1-1.33); renal failure (OR, 4.7; 95% CI, 1.52-14.41) and institution of parenteral nutrition (OR, 16.9; 95% CI, 1.07-269.03). Multivariate analysis showed the following risk factors for death in ICU: APACHE II scores (OR, 1.06; 95% CI, 1.01-1.11), and endotracheal intubation (OR, 5.07; 95% CI, 1.24-20.65). CONCLUSIONS: This study clearly documents a high prevalence rate of infections in the ICU, and the data suggest that occurrence of infections was associated with a prolonged ICU stay but had no significant effect on the final outcome. PMID- 16343638 TI - Aerobic capacity, oxidant stress, and chronic obstructive pulmonary disease--a new take on an old hypothesis. AB - Chronic obstructive pulmonary disease (COPD) is a smoking-related disorder that is a leading cause of death worldwide. It is associated with an accelerated rate of age-related decline in lung function due to the occurrence of destructive pathological changes such as emphysema, small airway remodeling, and mucus hypersecretion. Smokers are exposed to trillions of radicals and thousands of reactive chemicals and particles with every cigarette, thus oxidant stress is believed to be a central factor in the pathogenesis of COPD. The molecular activities of radicals, reactive oxygen, and nitrogen species can, over time, lead to a number of the detrimental changes in the lung. For instance, smoke can directly damage the mitochondrion, an organelle that has long been linked to age related diseases associated with oxidant stress. Mitochondria are involved in a number of important cellular processes and are the largest source of endogenous reactive oxygen species (ROS) in the cell; therefore, any impairment of mitochondrial function can lead to greater oxidant damage, cellular dysfunction, and eventually to disease. Only a subset of smokers (15-50%) develops COPD, suggesting that there are polygenetic and/or environmental susceptibility factors involved in this complex disease. Here, we propose that the aerobic capacity for an individual may determine whether one is susceptible to developing COPD. Aerobic capacity is a polygenetic trait closely associated with mitochondrial function, and we suggest antioxidant defenses. Thus, those smokers who have the greatest aerobic capacity will be most resistant to the effects of chronic cigarette smoke exposure and be less likely to develop COPD. PMID- 16343639 TI - Vascular endothelial dysfunction in diabetic cardiomyopathy: pathogenesis and potential treatment targets. AB - Cardiovascular complications account for significant morbidity and mortality in the diabetic population. Diabetic cardiomyopathy, a prominent cardiovascular complication, has been recognized as a microvascular disease that may lead to heart failure. Pathogenesis of diabetic cardiomyopathy involves vascular endothelial cell dysfunction, as well as myocyte necrosis. Clinical trials have identified hyperglycemia as the key determinant in the development of chronic diabetic complications. Sustained hyperglycemia induces several biochemical changes including increased non-enzymatic glycation, sorbitol-myoinositol mediated changes, redox potential alterations, and protein kinase C (PKC) activation, all of which have been implicated in diabetic cardiomyopathy. Other contributing metabolic abnormalities may include defective glucose transport, increased myocyte fatty acid uptake, and dysmetabolism. These biochemical changes manifest as hemodynamic alterations and structural changes that include capillary basement membrane (BM) thickening, interstitial fibrosis, and myocyte hypertrophy and necrosis. Diabetes-mediated biochemical anomalies show cross-interaction and complex interplay culminating in the activation of several intracellular signaling molecules. Studies in both animal and human diabetes have shown alteration of several factors including vasoactive molecules that may be instrumental in mediating structural and functional deficits at both the early and the late stages of the disease. In this review, we will highlight some of the important vascular changes leading to diabetic cardiomyopathy and discuss the emerging potential therapeutic interventions. PMID- 16343640 TI - Compatibility of glass-guided recording microelectrodes in the brain stem of squirrel monkeys with high-resolution 3D MRI. AB - Knowledge of the precise position of recording microelectrodes within the brain of a non-human primate is essential for a reliable exploration of very small anatomic structures. This work demonstrates the compatibility of a newly developed glass-guided microelectrode design and microfeed equipment with high resolution 3D magnetic resonance imaging (MRI). T1- and T2-weighted images allow for the non-invasive visualization of chronically implanted microelectrodes within the brain stem of squirrel monkeys in vivo. Neural extracellular multi unit recordings proved the functionality of the microelectrode before and after the use of 3D MRI suggesting the preservation of normal brain tissue at the tip of the electrode. Because histology confirmed the absence of lesions attributable to MRI, the approach offers an interactive monitoring during the course of neuroethological experiments. Consequently, MRI may become an in vivo alternative to common histological post mortem verifications of electrode tracks and hence may avoid the early sacrificing of primates after only a small number of experiments. PMID- 16343641 TI - Specificity of cognitive structure in depression and social phobia: a comparison of interpersonal and achievement content. AB - BACKGROUND: The comorbidity between the mood and anxiety is extensive and it is probable that individuals with these disorders share a number of cognitive characteristics. However, comorbidity rates with depression are not uniform among the various anxiety disorders. This study examined the common and distinguishing features of self-schematic structure in major depressive disorder and social anxiety compared to other anxiety disorders in general and to no psychiatric disturbance. METHODS: Participants completed the Psychological Distance Scaling Task, which provided an index of the cognitive organization of positively and negatively valenced interpersonal and achievement self-referent content. RESULTS: The depression and social phobia groups were statistically equivalent on all indices of cognitive organization, and showed greater interconnectedness among interpersonal negative content than both control groups, and less interconnectedness among both positive interpersonal and achievement content than did nonpsychiatric controls. Psychiatric groups were equivalent on negative achievement content but showed greater interconnectedness than controls. LIMITATIONS: This study had a modest sample size and the findings are constrained to females. CONCLUSIONS: Negative interpersonal content is more densely interconnected in individuals with social phobia and depression compared with both other anxiety disorder and nonpsychiatric controls. In addition, both social phobia and depression were associated with less interconnected positive content. These findings are indicative of similarities in the way self-schematic content may be organized in individuals with depression and social phobia. Theoretical explanations of these results and future research directions are discussed. PMID- 16343642 TI - Quantitative EEG analysis in dexamphetamine-responsive adults with attention deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (AD/HD) is recognized as a disorder affecting children and adolescents and has more recently been documented to continue into adulthood in a proportion of patients. One of the common treatments for AD/HD is the use of stimulant medications. Numerous studies have examined the therapeutic effects of stimulant medications in children, adolescents and adults with the disorder, reporting improvements in attention, concentration, and hyperactive and impulsive behaviours. Several studies have also examined the effects of stimulants on the electroencephalograph (EEG) of children and adolescents with the disorder, but to date, there have been no studies examining the effects of stimulant medication on the EEG of adults with AD/HD. In the present study, we aimed to replicate previous EEG findings in adults with AD/HD relative to controls and to examine whether there was any change in this profile following treatment with dexamphetamine. The EEG was recorded at rest in an eyes open condition from 50 adults diagnosed with AD/HD and assessed as good responders to treatment, both before and after treatment with dexamphetamine, and 50 control subjects. The pre-medication results are similar to those found in previous research that compared the EEGs of adults with AD/HD and control subjects. Following medication, there was a significant reduction in slow wave activity in the AD/HD group to levels similar to those in the control group. These results suggest that changes in brain function of good responders to dexamphetamine, as reflected in the EEG, may underlie the behavioural improvements observed in the clinical setting. PMID- 16343643 TI - Emotional information processing in violent patients with schizophrenia: association with psychopathy and symptomatology. AB - Schizophrenia and psychopathy have been independently shown to be associated with deficits in the recognition of facial expressions. These disorders are highly co morbid in forensic settings, and both are associated with aggressive behaviour. This study examines the relative contribution of psychopathic traits and psychotic symptoms to reported deficits in facial affect recognition in forensic patients with schizophrenia. Fifty-four male patients with schizophrenia were recruited from medium and high security hospitals. Participants were categorised into groups with high (HP), medium (MP) and low (LP) scores on the Psychopathy Checklist: Screening Version and based on symptomatology assessed using the Positive and Negative Syndrome Scale. Participants completed an animated facial affect recognition task assessing accuracy across the six basic emotions over high and low intensities. The HP group was found to have impaired recognition of sadness at low intensity compared with the LP group. In the overall sample, facial affect recognition for negatively valenced emotions was not related to positive or negative symptom scores. However, recognition accuracy for disgust was found to be negatively related to the severity of cognitive symptoms. Patients with high psychopathy scores and schizophrenia showed similar deficits in emotional information processing to those reported in the literature in non psychotic psychopathic samples. PMID- 16343644 TI - Dimensionality of the posttraumatic stress response among police officers and fire fighters: an evaluation of two self-report scales. AB - Confirmatory factor analyses were done to assess the dimensionality of the stress response in a sample of police officers and fire fighters (n = 1,168) involved in the 1992 air disaster in Amsterdam. The confirmatory factor analyses were applied to the responses on two psychometrically different instruments, i.e., the Self Rating Inventory for Posttraumatic Stress Disorder (SRIP) and the Impact of Event Scale (IES). The previously found distinction between (active) avoidance and numbing in samples highly affected by posttraumatic stress disorder appears to be applicable to the stress response of a less affected sample. For the SRIP, a five factor structure (i.e., intrusion, avoidance, hyperarousal, emotional numbing and sleep disturbance) appeared to fit slightly better than the four-factor structures from previous findings. For the IES, our results replicated findings of a four-dimensional structure (i.e., intrusion, avoidance, numbing and sleep disturbance) underlying the posttraumatic stress response. The factors of the best-fitting structure of both instruments proved reliable. Due to the psychometric properties of the two instruments, the relationship between similar factors in both instruments was only low to moderate. Compared with the IES, factors of the SRIP were, however, less discriminative from other symptoms of psychopathology. Replication in different traumatized or community samples is recommended. PMID- 16343645 TI - Decreased coherence in higher frequency ranges (beta and gamma) between central and frontal EEG in patients with schizophrenia: A preliminary report. AB - Schizophrenia is associated with a dysfunction of cognitive integration that may be due to abnormalities in inhibitory neural circuitry. A previous study found a failure of gamma band (25-45 Hz) synchronization in patients with schizophrenia compared to controls. Another recent study also stressed the importance of investigating high frequencies in the scalp-recorded sleep electroencephalogram (EEG). In this study, we compared coherence between first episode drug-naive patients with schizophrenia (n=8) and age- and sex-matched normal controls (n=8) using two 32-s epochs of C4 and F4 EEG. The coherence was obtained using 4096 data points (128 Hz signal) using cross-spectral analysis with Blackman-Tukey window in beta (15.25-24.75 Hz) and gamma (25-44.75 Hz) frequency bands. We used wake, non-rapid eye movement (NREM) and rapid eye movement (REM) sleep periods for the analyses. Our results show a significant decrease in coherence in both beta and gamma frequency bands in patients. Post-hoc 't' tests revealed a significantly lower coherence only during the wake stage in patients with schizophrenia in beta as well as gamma frequency bands. These results further support the importance of the analyses of high-frequency bands in the EEG and support previous findings of abnormal neural synchrony in patients with schizophrenia. These results have been discussed further in relation to wake and sleep stages. PMID- 16343646 TI - Occurrence of IgE in foals: evidence for transfer of maternal IgE by the colostrum and late onset of endogenous IgE production in the horse. AB - IgE is the key antibody involved in type I allergies. Allergen mediated crosslinking of IgE bound to high affinity Fcepsilon-receptors on mast cells and basophils stimulates cellular degranulation and release of inflammatory mediators and cytokines. In this report, we demonstrate that IgE antibodies can be transferred from the mother to offspring in horses via the colostrum. We found a clear correlation between the IgE concentration in colostrum and the total IgE concentration in foal sera on day 2 after birth (r(sp)=0.83). Maternal IgE was detected in foal sera by ELISA and on peripheral blood leukocytes of foals by flow cytometry. Both serum and cell membrane-bound IgE were undetectable in newborn foals before colostrum uptake and peaked on days 2-5 after birth. Cell bound IgE became undetectable at 2 months after birth. Serum IgE disappeared from the circulation within the first 3-4 months of age. These kinetics suggest that the IgE antibodies which are detectable in foals during the first 4 months after birth are of maternal origin only. The endogenous IgE production was found to begin at 9-11 months of age, when IgE could be detected on peripheral blood leukocytes and in foal sera again. After 18 months of life, the total IgE concentrations in foal sera were comparable to those detected in their dams. The late onset of endogenous IgE production offers an explanation for observations that IgE mediated allergies are generally not observed in horses before puberty. The roles of the passively transferred maternal IgE in newborn foals are not yet known, but could be manifold, ranging from passive immunity and induction of immunoregulatory functions to determinative influences of maternal IgE on the antibody repertoire in the offspring. PMID- 16343647 TI - Second branchial cleft fistula: is fistulogram necessary for complete excision. AB - We present our 10 years experience (1995-2005) of second branchial cleft fistulas and sinuses in the pediatric age group by retrospective analysis of records of 52 patients (with 63 fistulas) whose sinuses or fistulas were excised. It was found to be thrice as common in males as compared to females and was predominantly unilateral. The role of fistulogram and methylene blue dye injection in delineation and complete surgical excision of the tract was evaluated. Twenty five cases were managed by pre-operative fistulogram and intra-operative dye injections for excision, whereas 38 cases of branchial fistulas and sinuses were excised without fistulogram and dye injection. Fistulogram and dye injection were found to be of no extra help during excision of the tract. There were two recurrences. PMID- 16343648 TI - Pediatric cholesteatoma: a retrospective review. AB - OBJECTIVES: The optimal treatment for pediatric cholesteatoma is controversial. Management decisions including intact canal wall versus open cavity techniques, second look procedures and staging ossicular reconstruction continue to be debated. In an attempt to clarify this issue we conducted an 11-year retrospective analysis of our experience with cholesteatoma presenting in our pediatric population. STUDY DESIGN: Retrospective review of children undergoing surgical intervention for cholesteatoma at a tertiary care pediatric hospital between 1 July 1992 and 1 July 2003 by the senior author. METHODS: Comparison of recurrence rates in intact canal wall (ICW) versus canal wall down (CWD) procedures; with analysis of second-look procedures, ossicular chain reconstruction (OCR) and hearing results in the management of pediatric cholesteatoma. RESULTS: Two hundred and sixty-two children with 278 cases of cholesteatoma underwent surgical resection. Of these children, 221 were managed via an ICW approach while the remaining 57 underwent a CWD procedure. The overall recurrence rate in this series was 16%, with 17% in the ICW group and 12% in the CWD group. OCR was performed in 97% of the ICW cases at time of second-look procedure, with 75% undergoing reconstruction with partial ossicular reconstruction prosthesis. The average air-bone gap improvement in these patients was 10.8 dB, with an average hearing improvement of total ossicular reconstruction of 5.8 dB. The average hearing improvement in the CWD group, all managed with cartilage interposition grafts, was 3.7 dB. CONCLUSIONS: Management of pediatric cholesteatoma requires a highly individualized approach that takes into account anatomic, clinical and social factors to determine the most successful surgical treatment paradigm. PMID- 16343649 TI - Quantitation of HIV-1 RNA in breast milk by real time PCR. AB - HIV-1 RNA in breast milk is a strong predictor of HIV-1 transmission through breastfeeding. In the present report, breast milk samples from HIV-1 uninfected donors were spiked with dilution of quantified culture supernatant from HIV 1(NDK) infected PBMC. Two RNA extraction techniques based on silica extraction, Nuclisens (BioMerieux) and Triazol (Qiagen), two techniques based on guanidine thiocynanate/chloroforme extraction, TRIzol (Life Technologie) and Amplicor HIV-1 Monitor (Roche Diagnostic Systems), and one technique based on electrostatic adsorption on iron oxide micro beads (Promega) were compared. HIV-1 RNA was quantitated by real time PCR (LTR gene) and Amplicor HIV-1 Monitor. Combining magnetic micro beads extraction and real time PCR quantitation allowed to correctly quantify breast milk HIV-1 RNA, with a difference between the expected and measured HIV-1 RNA levels always lower than 0.3 log copies/ml. The same combination was confirmed on 25 breast milk samples from HIV-1 infected women collected in Kwazulu-Natal, South Africa, by comparing measurements with those obtained by the Amplicor HIV-1 Monitor (r(2)=0.88). Nucleic acid extraction by magnetic micro beads followed by real time PCR is a reliable, sensitive, rapid and simple procedure to quantify HIV-1 RNA in breast milk and allows for PCR inhibitors found frequently in these samples. PMID- 16343650 TI - Novel one-step real-time RT-PCR assay for rapid and specific diagnosis of Crimean Congo hemorrhagic fever encountered in the Balkans. AB - Early and accurate diagnosis of Crimean-Congo hemorrhagic fever (CCHF) is essential for the treatment and outcome of the disease and prevention of its further transmission. Molecular-based diagnostic assays now serve as the front line tool in the diagnosis of CCHF. However, the development of real-time RT-PCR assay for the detection of Crimean-Congo hemorrhagic fever virus (CCHFV) has been hampered by a virus strain variation. The development of a one-step real-time RT PCR assay for the detection of CCHFV is described herein. The technique is based on the fluorescence resonance energy transfer probe technology employing the endonuclease activity of Taq polymerase enzyme. The assay was designed to detect specifically the strains from a phylogenetic cluster of CCHFV which encompasses the known CCHFV strains circulating in the Balkan region. The detection system was tested using CCHFV strain Kosovo Hoti, clinical serum samples and ticks. The real-time assay described is rapid, specific and sensitive. Since the Balkan peninsula is also an endemic region for hemorrhagic fever with renal syndrome (HFRS), this method is suggested as convenient for early differential diagnosis of suspected viral hemorrhagic fever patients. PMID- 16343651 TI - Identification and characterization of potent small molecule inhibitor of hemorrhagic fever New World arenaviruses. AB - Category A arenaviruses as defined by the National Institute of Allergy and Infectious Diseases (NIAID) are human pathogens that could be weaponized by bioterrorists. Many of these deadly viruses require biosafety level-4 (BSL-4) containment for all laboratory work, which limits traditional laboratory high throughput screening (HTS) for identification of small molecule inhibitors. For those reasons, a related BSL-2 New World arenavirus, Tacaribe virus, 67-78% identical to Junin virus at the amino acid level, was used in a HTS campaign where approximately 400,000 small molecule compounds were screened in a Tacaribe virus-induced cytopathic effect (CPE) assay. Compounds identified in this screen showed antiviral activity and specificity against not only Tacaribe virus, but also the Category A New World arenaviruses (Junin, Machupo, and Guanarito). Drug resistant variants were isolated, suggesting that these compounds act through inhibition of a viral protein, the viral glycoprotein (GP2), and not through cellular toxicity mechanisms. A lead compound, ST-294, has been chosen for drug development. This potent and selective compound, with good bioavailability, demonstrated protective anti-viral efficacy in a Tacaribe mouse challenge model. This series of compounds represent a new class of inhibitors that may warrant further development for potential inclusion in a strategic stockpile. PMID- 16343652 TI - Amphetamine-induced 50 kHz calls from rat nucleus accumbens: a quantitative mapping study and acoustic analysis. AB - Emission of 50 kHz ultrasonic calls in rats is known to be associated with appetitive behavioural situations and positive social interactions. The purpose of the study was to pharmacologically characterize amphetamine-induced 50 kHz calls and to perform quantitative mapping of this response in the nucleus accumbens. Injections of amphetamine into the nucleus accumbens induced species typical 50 kHz calls in adult rats. The acoustic parameters of the calls were not affected by different amphetamine doses or combination of agents. The increase in the number of calls occurred predominantly from the accumbens shell and to a lesser degree from the core region. This effect was dose-dependent within the range of 1-20 microg of amphetamine and was reversed by pretreatment with D1 or D2 dopamine antagonists (SKF-83566 or raclopride) administered to the same brain site. However, another D2 dopamine receptor antagonist, haloperidol, which is known to increase the accumbens dopamine level, was ineffective in reversing the increase in call number at the dose studied. On the contrary, intraacumbens haloperidol, when injected alone, caused an increase in 50 kHz calls. It is concluded that the release of dopamine, predominantly in the accumbens shell region, is responsible for production of 50 kHz calls and the calls may indicate an appetitive state compatible with anticipation of reward and positive affect. Both D1 and D2 subtypes of dopamine receptors may be necessary to induce 50 kHz calls and signal the appetitive state. PMID- 16343653 TI - Social behavior deficits in the Fmr1 mutant mouse. AB - Mice exhibiting deficits in social behavior may provide valuable models for autistic-like behavioral problems. We tested social interactions in male mice from three inbred strains: C57BL/6J (B6), BALB/cJ (C) and DBA/2J (D2). All three strains showed gradual habituation of the number of social interactions with an ovariectomized female over four subsequent 2min sessions, returning to initial levels when presented with another stimulus mouse. Next, we studied males with a knockout mutation in the Fmr1 gene on a B6 background. KO animals showed strongly reduced levels of social interaction, which were about similar as those of habituated controls. This social behavior deficit suggests that Fmr1 KO mice could possibly be used as models for autistic behaviors. PMID- 16343654 TI - Neurotrophin levels and behaviour in BALB/c mice: impact of intermittent exposure to individual housing and wheel running. AB - This study assessed the effects of intermittent individual housing on behaviour and brain neurotrophins, and whether physical exercise could influence alternate individual-housing-induced effects. Five-week-old BALB/c mice were either housed in enhanced social (E) or standard social (S) housing conditions for 2 weeks. Thereafter they were divided into six groups and for 6 weeks remained in the following experimental conditions: Control groups remained in their respective housing conditions (E-control, S-control); enhanced individual (E-individual) and standard individual (S-individual) groups were exposed every other day to individual cages without running-wheels; enhanced running-wheel (E-wheel) and standard running-wheel (S-wheel) groups were put on alternate days in individual running-wheel cages. Animals were assessed for activity in an automated individual cage system (LABORAS) and brain neurotrophins analysed. Intermittent individual housing increased behavioural activity and reduced nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) levels in frontal cortex; while it increased BDNF level in the amygdala and BDNF protein and mRNA in hippocampus. Besides normalizing motor activity and regulating BDNF and NGF levels in hippocampus, amygdala and cerebellum, physical exercise did not attenuate reduction of cortical NGF and BDNF induced by intermittent individual housing. This study demonstrates that alternate individual housing has significant impact on behaviour and brain neurotrophin levels in mice, which can be partially altered by voluntary physical exercise. Our results also suggest that some changes in neurotrophin levels induced by intermittent individual housing are not similar to those caused by continuous individual housing. PMID- 16343655 TI - Spontaneous alternation behaviour in rats: kynurenic acid attenuated deficits induced by MK-801. AB - The present study was undertaken to investigate the effects of pharmacological modulation of the NMDA receptors on spontaneous alternation behaviour. The performance of rats treated with MK-801 and kynurenic acid (KYNA) was assessed in the cross-arm-maze. We evaluated: (a) the total number of arm entries representing locomotor activity, (b) spontaneous variation of different arms thought to reflect alternation performance. In the first experiment, MK-801 (0.01, 0.025, 0.05, 0.1 and 0.2 mg/kg, i.p.) was given 30 min prior to the testing. Beginning the dose of 0.05 mg/kg the drug increased locomotion and impaired alternation performance. An ability of animals to enter subsequently three or four different arms was reduced significantly. In the second experiment, the dose of 0.05 mg/kg was chosen as the lowest possible dose of MK-801 producing marked behavioural impairment. KYNA (0.3, 3 and 30 mg/kg, s.c.) was administered 60 min prior to the MK-801. While all KYNA doses prevented hyperlocomotion, only the highest dose (30 mg/kg) maintained alternation score at the control levels, i.e. the KYNA plus MK-801 treated animals alternated regularly three or four different arms. The results suggest different sensitivity of the two behavioural systems, i.e. locomotion and space orientation, towards pharmacological insult. In conclusion, the study confirmed protective behavioural effects of KYNA given in sufficient amounts and sufficiently long prior MK-801. PMID- 16343656 TI - Brain antioxidant levels in hamsters during hibernation, arousal and cenothermia. AB - Warming from hibernation to cenothermia involves intense metabolic activity coincident with large fluxes in blood flow and is considered to be a period of oxidative stress during which utilization of endogenous antioxidants prevents pathology. Very slow flow brain microdialysis enabled temperature independent sampling of the brain striatal extracellular fluid (ECF) during hibernation, arousal and cenothermia in Syrian hamsters (Mesocricetus auratus). Brain tissue and dialysates were analyzed to provide the first profile of changes in ECF levels of ascorbate (AA), glutathione (GSH) and urate during hibernation and the transition to cenothermia. Brain tissue content of AA and GSH was unchanged between hibernation and cenothermia; however, arousal was associated with substantial oxidation of AA from the brain ECF and plasma compartments. ECF GSH increased during arousal. Brain tissue urate content was decreased 50% during hibernation. ECF urate levels were unchanged in hibernation and cenothermia but transiently increased 100% during arousal. These experiments demonstrate that arousal from hibernation is a suitable experimental model for examination of the mechanisms by which non-pathological tissue integrity is maintained in the face of the generation of free radicals during increasing metabolism, temperature and cerebral reperfusion. PMID- 16343657 TI - Quantitative changes in metallothionein expression in target cell-types in the gills of turbot (Scophthalmus maximus) exposed to Cd, Cu, Zn and after a depuration treatment. AB - Turbot (Scophthalmus maximus) were exposed to two sublethal concentrations (1 and 10 mg metal/l) of cadmium (8.9 and 89 microM Cd), copper (15.26 and 152.6 microM Cu) and zinc (15.3 and 153 microM Zn) for 7 days, and afterwards were maintained depurating for 14 days. Immunoreactive metallothioneins (irMTs) and metal ions were localized in the branchial epithelium by immunohistochemistry (using an anti Cod MT antibody) and autometallography (AMG), respectively. Metal ions were demonstrated by AMG as black silver deposits (BSD), mainly in mucocytes (MC) and to a lesser extent in the other branchial cell-types (respiratory cells (RC), chloride cells (CC) and basal layer cells (BLC)). Irrespective of the metal supplied, BSD were rapidly visualized in MC after 1 h of exposure. This accumulation did not increase with increasing exposure time and concentration. Metallothionein expression was mainly observed in mature CC in the interlamellar space for all exposure conditions and it was shown that all mature cells express the same amount of irMT. The number of CC exhibiting irMT in metal-exposed turbots increased following short exposure times (1 h-1 day) in the filament epithelium and following longer exposure times (1-7 days) in the secondary lamellae. Total levels of irMT in the gills (quantified by image analysis and densitometry) increased significantly in metal-exposed turbot and were related to increased exposure times. It can be concluded that the total content of irMT in the gills of metal-exposed turbot is governed by changes in the number of mature CC expressing the protein. The quantification of total irMT in branchial CC can be considered as a reliable biomarker of metal exposure since reflects changes in metal bioavailability. This approach based on cell-selective immunohistochemistry can be simplified by only quantifying the number of mature CC. In addition, the dramatic increase of CC in the gills that produces epithelial thickening of the FE enhances migration of CC up to the edge of the SL and provokes the hypertrophy and fusion of secondary lamellae can be considered as unspecific biomarkers of effect indicating disturbed health in turbot. PMID- 16343658 TI - The role of Trypanosoma brucei MRPA in melarsoprol susceptibility. AB - We previously showed that over-expression of Trypanosoma brucei MRPA, a member of the multidrug resistance protein family in T. brucei, reproducibly resulted in resistance to the anti-trypanosomal drug melarsoprol in vitro. MRPA is predicted to mediate efflux of melarsoprol as a conjugate with trypanothione, a glutathione spermidine conjugate which is the major small thiol in trypanosomes. Here, we show that depletion of MRPA by RNA interference resulted in moderate hypersensitivity to both melarsoprol and melarsen oxide. Over-expression of MRPA alone is not sufficient to cause melarsoprol resistance in vivo, although it is sufficient in vitro. This discrepancy is not an effect of drug metabolism since over-expression of MRPA alone conferred resistance to melarsoprol and its principle metabolite, melarsen oxide, in vitro. Over-expression of MRPA was not detected in four melarsoprol-resistant trypanosome isolates from sleeping sickness patients. PMID- 16343659 TI - Transcriptional regulation of the Drosophila orc2 gene by the DREF pathway. AB - DNA replication-related element (DRE) and the DRE-binding factor (DREF) play an important role in regulating DNA replication-related genes such as PCNA and DNA polymerase alpha in Drosophila. We have previously reported that overexpression of DREF in developing eye imaginal discs induced ectopic DNA synthesis and apoptosis, which results in rough eyes. To identify genetic interactants with the DREF gene, we have carried out a screen for modifiers of the rough eye phenotype. One of the suppressor genes identified was the Drosophila orc2 gene. A search for known transcription factor recognition sites revealed that the orc2 gene contains three DREs, named DRE1 (+14 to +21), DRE2 (-205 to -198), and DRE3 (-709 to 702). Band mobility shift analysis using Kc cell nuclear extracts detected the specific complex formed between DREF and the DRE1 or DRE2. Specific binding of DREF to genomic region containing the DRE1 or DRE2 was further demonstrated by chromatin immunoprecipitation assays, suggesting that these are the genuine complexes formed in vivo. The luciferase assay in Kc cells indicated that the DRE sites in the orc2 promoter are involved in a transcriptional regulation of the orc2 gene. The results, taken together, demonstrate that the orc2 gene is under the control of DREF pathway. PMID- 16343660 TI - Stent graft implantation for in-stent restenosis of coronary artery stenosis after Kawasaki disease. AB - Coronary artery stenosis is a major complication of Kawasaki disease (KD). Several interventional methods in treating coronary artery stenosis have been introduced. However, there are few reports on the management of in-stent restenosis after coronary stent implantation in children. Reported is a 10-year old boy who underwent successful stent graft insertion for treating in-stent restenosis with neoaneurysm formation after stent implantation for severe coronary stenosis after KD. Twenty-eight months follow-up studies showed no significant restenosis and perfusion defect. PMID- 16343661 TI - The heart in renovascular disease--an association demanding further investigation. AB - A close relationship exists between cardiovascular and renal disease; they often occur concomitantly, and abnormalities in either system are pathophysiologically important in both causing disease and determining clinical outcome in the other. Whilst the main focus of the article relates to the adverse association between atherosclerotic renovascular disease (ARVD) and the cardiovascular system, it is important to briefly review relevant epidemiology. PMID- 16343662 TI - Current possibilities for detecting high risk of cardiovascular disease. AB - Current possibilities for better detecting high risk of coronary heart disease (CHD) and stroke and peripheral arterial disease are described in this review. A first step is based on risk factors assessment that allows establishing high-risk diagnostic, either by detecting a condition termed as "CHD risk equivalent" and defined by one or more severe major risk factor, or by calculating multifactorial risk in asymptomatic subjects with a global risk score integrating several moderate risk factors. A second diagnostic step, concerning subjects not considered at high-risk by risk factors assessment, is based on non-invasive detection of sub clinical atherosclerosis via a wide variety of structural and functional arterial markers. A third step focuses on detection of myocardial ischemia that may add diagnostic and prognostic information in subjects with high CHD risk. The implementation of high-risk strategy is not yet standardized but it should allow improving cost-effectiveness of cardiovascular prevention, particularly in asymptomatic subjects. PMID- 16343663 TI - Big endothelin-1 is not a predictor in aortic stenosis, but is related to arterial blood pressure. AB - BACKGROUND: In aortic stenosis, natriuretic peptides have recently been shown to correlate with ventricular function and to predict symptom-free survival and outcome. Elevated big endothelin-1 (bigET) is associated with poor prognosis in chronic heart failure, but little is known about its role in severe aortic stenosis. METHODS: In 61 patients with aortic stenosis (71+/-10 years, mean gradient 65+/-20 mm Hg, valve area 0.63+/-0.15 cm2), plasma bigET was determined by radioimmunoassay and related to echocardiographic parameters, symptoms and survival. Patients were followed for 1 year. RESULTS: BigET (mean 2.3+/-1.5, range 0.1-7.5 fmol/ml) was elevated > or = 1.9 fmol/ml in 54% of patients, but was not correlated to the transvalvular gradients or valve area. BigET did not differ significantly between 14 asymptomatic (2.4+/-1.0 fmol/ml) and 47 symptomatic patients (2.3+/-1.6 fmol/ml), although the highest levels were observed in 5 patients in NYHA class III-IV (4.2+/-2.2 fmol/ml, p=0.035). No significant difference in bigET was observed between 51 survivors and 10 patients who died during follow-up (2.2+/-1.4 vs 2.7+/-1.6 fmol/ml). BigET did not differ between 7 asymptomatic patients developing symptoms and those remaining asymptomatic during follow-up. BigET was significantly related to the systolic blood pressure and left ventricular systolic pressure (r=0.389, p=0.0025 and r=0.401, p=0.0018, respectively), but not to the diastolic blood pressure or interventricular septal wall thickness. BigET was inversely related to the left ventricular ejection fraction (r=0.327, p=0.01) and fractional shortening (r=0.391, p=0.044). CONCLUSION: Although frequently elevated, bigET-1 is not a useful predictor of symptoms or outcome in patients with severe aortic stenosis. BigET increases inversely with left ventricular function and directly with systolic left ventricular and blood pressure, but is not related to transvalvular gradients or valve area. PMID- 16343664 TI - Optimal antiplatelet treatment for percutaneous coronary intervention: clopidogrel vs. ticlopidine. AB - Antiplatelet treatment for patients undergoing percutaneous coronary interventions is a rapidly changing area. Thienopyridines derivatives (ticlopidine and clopidogrel) have shown to decrease major cardiovascular events. Ticlopidine can cause rare but serious side effects, especially during the first 3 months of treatment. Clopidogrel appears to be a safer alternative to ticlopidine. However, resistance to clopidogrel therapy may increases the risk of recurrent cardiovascular events. Whether increased doses of clopidogrel might overcome this resistance in nonresponsive patients warrants further investigation. PMID- 16343665 TI - Myocardial infarction in isolated ventricular non-compaction: contrast echo and MRI. AB - We describe the case of an occasional discovery of isolated ventricular non compaction in an adult recovered for an acute myocardial infarction, in which only the echocardiogram revealed an isolated ventricular non-compaction, confirmed by MRI: an unusual association between coronary artery disease and isolated ventricular non-compaction. PMID- 16343666 TI - Antiplatelet versus anticoagulant therapies in advanced age: an unfinished task. AB - We have read with great interest a retrospective cohort study recently published by Blich and Gross. In our opinion, this article renews the controversy of the best antithrombotic therapy in patients with AF. The use of anticoagulant treatment to prevent the occurrence of stroke in patients with AF is supported by several randomized controlled clinical trials. Aspirin is also effective in preventing stroke in AF, but both direct and indirect comparisons with oral anticoagulation suggest less effectiveness. However, very probably these patients are quite different than those seen in the clinical practice. The role of antiplatelet therapy is not completely established, and the selection between aspirin or warfarin in advanced age remains an unfinished task. PMID- 16343667 TI - Advanced oxidation protein products (AOPP) and high-sensitive C-reactive protein (hs-CRP) in an "atheroma-free model": Down's syndrome. AB - Murdoch et al. in 1977 called Down syndrome an "atheroma-free model." In this preliminary study, we investigated advanced oxidation protein products (AOPP) and high-sensitive C-reactive protein (hs-CRP) in 47 age-matched Down syndrome patients and 20 age-matched healthy controls. In these healthy patients, we detected no new biochemical risk factors for atherosclerosis, such as AOPP and hs CRP, so risks are probably considerably lower. PMID- 16343668 TI - Socioeconomic factors in the prognosis of heart failure in a Brazilian cohort. AB - A large body of evidence supports the concept that socioeconomic factors may influence the prognosis of cardiovascular diseases. We studied the influence of socioeconomic factors in the prognosis of heart failure in 494 patients consecutively admitted to the hospital due to worsening of symptoms of heart failure in Sao Paulo, Brazil. The ages ranged between 15 and 90 (mean 57.5, standard deviation 14.2) years; 345 (69.8%) were men and 149 (30.2%) were women. Patients were followed-up at least for two years after hospital discharge. Two hundred and forty-one patients died due to cardiac disease during the follow-up period. Hierarchical analysis was performed with demographic and socioeconomic variables analyzed in the first level and clinical variables in the second level. At the level of the demographic and socioeconomic characteristics, care under the Public Health System (odds ratio 3.46, 95% CI 1.91 to 6.27, p<0.001) was the most important predictive factor for mortality. At the second hierarchical level, the predictive factors of mortality were Chagas' heart disease (odds ratio 3.34, 95% CI 1.90 to 5 89, p<0.001), digoxin therapy (odds ratio 1.94, 95% CI 1.09 to 3.45, p<0.025), frequency of myocardial infarctions (odds ratio 1.75, 95% CI 1.25 to 2.44, p<0.001) and arterial hypertension (odds ratio 0.47, 95% CI 0.30 to 0.73, p=0.001). Care delivered through the Public Health System may be a marker of socioeconomic conditions related with higher mortality in patients with heart failure. PMID- 16343669 TI - Herd-level risk factors for Neospora caninum seroprevalence in dairy farms in southern Brazil. AB - A cross-sectional study was used to test the relationship between herd seroprevalence to Neospora caninum and various potential herd-level risk factors in 60 dairy farms located in two distinct regions in southern Brazil. Thirty farms were randomly selected from within each region. A questionnaire was designed to summarize each farm's production system as it might relate to N. caninum transmission. The questionnaire contained 105 closed questions relating to general characteristics of the farms, farm facilities, management, source of food and water, herd health, environment and biosecurity, which included questions relevant to N. caninum transmission, including presence and number of dogs and other animals, purchase of animals and contact with man. Serum samples were collected from 40% of animals in each farm and N. caninum antibodies were detected by immunofluorescent antibody test (IFAT). The association between potential risk factors and the probability of an animal being seropositive was modeled using a generalized estimation equations (GEE) logistic regression model. The model accounted for multilevel correlation of data from multiple animals within herds. The mean (+/-S.D.) number of animals in the 60 herds was 64.5 (+/ 45.6), ranging from 20 to 280 females. Blood samples were collected from 1549 animals. The size of the farms varied from 4 to 100 ha (mean 30.1+/-25.9 ha). At least one dog was found in 57 of the 60 dairy farms (95%). The mean number of dogs was 3.1 (+/-1.9), ranging from 0 to 10. All females were raised on pasture. For all cattle sampled, N. caninum seroprevalence was 17.8%. Overall, 93.3% of herds (56/60) had at least one seropositive animal identified. Four variables were significantly associated with N. caninum sero-response in the 57 dairy farms, which were included in the final multivariable model: the number of dogs on the farm, farm area (hectares), feeding pooled sources of colostrum and region. The odds of a cow being seropositive increased 1.13 times for each additional dog present on the farm (P=0.021). Cattle from farms that fed calves colostrum pooled from multiple cows had 1.79 times greater odds for being seropositive for N. caninum (P<0.003). The probability of being seropositive was inverse to the area of the farms, such that cattle had 0.92 times the odds to be seropositive (P=0.014) for each additional 10 ha of farmland. Finally, cattle from farms in region one had 0.71 times the odds to be seropositive than cattle from region two (P=0.035). Results of this study suggest that several risk factors may explain why dairy cattle in Brazil may become exposed to N. caninum. However, further investigation of these factors is necessary because the purpose of this study was to refine and generate hypotheses on N. caninum transmission. PMID- 16343670 TI - Early initiation of highly active antiretroviral therapies for AIDS: dynamic choice with endogenous and exogenous learning. AB - Criteria for initiation of highly active antiretroviral treatments (HAART) in HIV infected patients remain a matter of debate world-wide because short-term benefits have to be balanced with costs of these therapies, and restrictions placed on future treatment options if resistant viral strains develop. On the other hand, postponing the introduction of HAART may involve a therapeutic opportunity cost if a patient's health is allowed to deteriorate to such an extent of becoming unable to benefit from new treatments currently under development when they become available. We introduce a two period model where period one treatment adoption is an irreversible act with future, but uncertain, consequences. New information, both endogenous and exogenous, arises over time and shapes the conditions surrounding the second period therapeutic decision. A surprising result is that, under conditions that appear close to those surrounding the HAART debate, the magnitude of the feared resistance effect has no effect on leaves the optimal treatment decision as far as it is high enough. PMID- 16343671 TI - Primers containing universal bases reduce multiple amoA gene specific DGGE band patterns when analysing the diversity of beta-ammonia oxidizers in the environment. AB - The gene encoding the active site of the ammonia monooxygenase (amoA) has been exploited as molecular marker for studying ammonia-oxidizing bacteria (AOB) diversity in the environment. Primers amplifying functional genes are often degenerated and therefore produce multiple band patterns, when analysed with the Denaturing gradient gel electrophoresis (DGGE) approach. To improve the DGGE band patterns we have designed new primer sets which contain inosine residues and are specific for the amoA gene. Primers were evaluated analysing pure AOB cultures and two habitats (wastewater treatment plant, soda pools). We found that the application of inosine primers helped to reduce the apparent complexity of the DGGE band pattern. Comparison of sequences from environmental samples using either degenerated or inosine containing amoA primers retrieved both identical and additional sequences. Both primer sets seem to be limited in their ability to detect the presence of all AOB by DGGE analyses. PMID- 16343672 TI - Circadian variability is negligible in primary visual cortices as measured by fNIRS. AB - Neural activation leads to an increase of regional cerebral blood flow. Most of the functional imaging studies implicitly assume that variability of the hemodynamic response throughout a single day is negligible. To test this assumption we measured brain activation by functional near-infrared spectroscopy (fNIRS) in the visual cortex of ten subjects six times throughout the day, from 0800-1800 h, during an event-related checkerboard paradigm. Concentration of oxygenated hemoglobin increased, whereas concentration of deoxygenated hemoglobin decreased at each time point examined, without significant influences of daytime. Variability of the hemodynamic response was higher across subjects than for single subjects across day. In conclusion, our study is the first one supporting the common practice of ignoring circadian variability in functional imaging studies. PMID- 16343673 TI - Interlimb coordination following stroke. AB - Studies investigating whether simultaneous bilateral movements can facilitate performance of the impaired limb(s) of stroke patients have returned mixed results. In the present study we compared unilateral limb performance (amplitude, cycle duration) with performance during an interlimb coordination task involving both homologous (both arms, both legs) and non-homologous (one arm, one leg) limbs in stroke participants (n=7) and healthy age-matched controls (n=7). In addition, the effect of on-line augmented visual feedback on interlimb coordination was investigated. Participants performed cyclical flexion-extension movements of the arms and legs in the sagittal plane paced by an auditory metronome (1 Hz). Movement amplitudes were larger and cycle durations shorter during homologous limb coordination than non-homologous coordination. Compared with unilateral movements both groups had reduced movement amplitudes and the stroke group increased cycle duration when interlimb coordination tasks were performed. These effects were most evident during non-homologous (arm and leg) coordination. No evidence of facilitation of the impaired limb(s) was found in any of the interlimb coordination conditions. Augmented visual feedback had minimal effect on the movements of control participants but lead to an increase of cycle duration for stroke participants. PMID- 16343674 TI - Envelope gene capture and insect retrovirus evolution: the relationship between errantivirus and baculovirus envelope proteins. AB - In this report the evolution of insect retroviruses (errantiviruses) is reviewed with particular emphasis on the relationship between their env protein and a baculovirus envelope fusion protein. In addition, selected features of the env protein from the errantivirus Dme17.6V are examined. These include characterization of the 21 amino acid predicted fusion peptide sequence that is highly homologous to a region of baculovirus envelope fusion proteins. We found that, although this sequence could not substitute for the homologous sequence in the baculovirus LD130 envelope fusion protein, by changing four amino acids, the hybrid construct became active for low-pH induced cell fusion. In addition, a Dme17.6V env-egfp construct was found to localize to cell membranes. PMID- 16343675 TI - The intron 1 of HPV 16 has a suboptimal branch point at a guanosine. AB - The branch point sequence (BPS) of intron 1 of the HPV-16 was determined via RT PCR in a cell free system, using lariat intermediates obtained by in vitro splicing reactions. We used synthetic E6/E7 transcripts and HeLa nuclear protein extracts to obtain the splicing intermediates. Then, a divergent oligonucleotide primer set, pairing on the lariat RNA that encompassed the 2'-5' phosphodiester bond formed between the 5' end of the intron and the BPS, was used for cDNA synthesis and PCR amplification. Subsequent RT-PCR assays revealed four splicing intermediates, made up of a major intermediary corresponding to the BPS and four cryptic branched sequences. Only intermediates bound at the 5' end of the intron are probably the authentic branch point sequence, and all of them branch at guanosine 328 instead of the typical adenosine. Unusually, the BPS of intron 1 of HPV-16 is a suboptimal sequence (AGUGAGU) that differs from the eukaryotic consensus BPS, which correlates with the splicing profile observed for early transcripts of HPV-16 in tumors and tumor derived cell lines. The implications of this unusual branch point sequence for splicing of the HPV-16 pre-mRNA are discussed. PMID- 16343676 TI - Mayaro virus: complete nucleotide sequence and phylogenetic relationships with other alphaviruses. AB - Mayaro (MAY) virus is a member of the genus Alphavirus in the family Togaviridae. Alphaviruses are distributed throughout the world and cause a wide range of diseases in humans and animals. Here, we determined the complete nucleotide sequence of MAY from a viral strain isolated from a French Guianese patient. The deduced MAY genome was 11,429 nucleotides in length, excluding the 5' cap nucleotide and 3' poly(A) tail. Nucleotide and amino acid homologies, as well as phylogenetic analyses of the obtained sequence confirmed that MAY is not a recombinant virus and belongs to the Semliki Forest complex according to the antigenic complex classification. Furthermore, analyses based on the E1 region revealed that MAY is closely related to Una virus, the only other South American virus clustering with the Old World viruses. On the basis of our results and of the alphaviruses diversity and pathogenicity, we suggest that alphaviruses may have an Old World origin. PMID- 16343677 TI - Importance of the extracellular and cytoplasmic/transmembrane domains of the haemagglutinin protein of rinderpest virus for recovery of viable virus from cDNA copies. AB - A specific interaction between the F and H proteins is required to enable fusion of the virus and host cell membranes and in some cases these proteins are not interchangeable between related viruses of the family Paramyxoviridae. For example, the F and H proteins of two ruminant morbilliviruses, rinderpest virus (RPV) and Peste-des-petits-ruminants virus (PPRV), are not interchangeable since viable virus could not be rescued from cDNA constructs where an individual glycoprotein gene of RPV was replaced with that from PPRV. To investigate which domain of the H protein, extracellular or cytoplasmic/transmembrane, was most important for preventing this interaction, two chimeric H gene constructs were made where the normal H gene of RPV was substituted with variant H genes where the transmembrane/cytoplasmic tail region (pRPV2C-PPRTm) or the whole ectodomain (pRPV2C-PPRExt) were derived from PPRV. Chimeric viruses were rescued from both the constructs and, while RPV2C-PPRTm virus grew to as high titres as the parent virus, RPV2C-PPRExt virus was extremely debilitated with respect to growth in tissue culture. Thus the ectodomain of H is the most important region required for effective interactions of the two glycoproteins for the recovery of viable virus. Nevertheless, the transmembrane/cytoplasmic domain of RPV alone can allow a chimeric virus to be rescued, which was not possible when the complete H gene was derived from PPRV. Both versions of the H protein and also the F protein were found to be incorporated into the envelope of the budded virions. PMID- 16343678 TI - Iontophoresis: a non-invasive ocular drug delivery. AB - Iontophoresis as a non-invasive technique for ocular drug delivery has been investigated for many years. This paper provides an overview of the approaches currently used in the development of the ocular iontophoretic device, the essential features of this procedure and the reported toxicity. This review focuses on the experimental results after transcorneal and transscleral iontophoresis of different drugs, emphasizing the current density applied and the treatment duration used by the investigators. PMID- 16343679 TI - Primary prophylaxis with nadolol in cirrhotic patients: Doppler patterns of splanchnic hemodynamics in good and poor responders. AB - BACKGROUND/AIMS: We aimed to characterize by echo-color-Doppler the splanchnic hemodynamics of patients good and poor responders to primary prophylaxis with nadolol. METHODS: Thirty cirrhotic patients (Child-score 7.0+/-1.8) with medium/large esophageal varices without previous bleedings were consecutively enrolled. At inclusion and after 3 months of treatment with nadolol, they underwent a splanchnic echo-color-Doppler study and a measurement of hepatic venous pressure gradient (HVPG). RESULTS: Nadolol (60+/-36 mg/day; range 20-160) induced a significant reduction of HVPG (16.6+/-6.1 vs. 19.4+/-4.6 mmHg, P < 0.0001). 13 patients (43.3%) were hemodynamic responders. Responders and Poor responders had similar baseline clinical characteristics. Poor-responders at baseline were characterized by lower impedance indexes in superior mesenteric artery (SMA) (PI 2.29+/-0.45 vs. 2.74+/-0.46; P = 0.01; RI 0.83+/-0.04 vs. 0.86+/ 0.03; P = 0.02), hepatic artery (HA) (PI 1.41+/-0.19 vs. 1.79+/-0.48; P = 0.03; RI 0.71+/-0.05 vs. 0.80+/-0.07; P = 0.02), and splenic artery (SA) (PI 1.18+/ 0.27 vs. 1.73+/-0.40; P = 0.01; RI 0.66+/-0.07 vs. 0.73+/-0.09; P = 0.02), and by higher mean flow velocity of HA (52.6+/-21.6 vs. 26.5+/-9.5 cm/s; P = 0.02) and SMA (49.7+/-14.5 vs. 33.9+/-13.1 cm/s; P = 0.06). CONCLUSIONS: Cirrhotic patients poor-responders to nadolol show a pronounced arterial splanchnic vasodilatation at a baseline echo-color-Doppler study. This can be considered a non-invasive clue for the a priori identification of this subgroup of patients. PMID- 16343680 TI - Significance of liver negative-strand HCV RNA quantitation in chronic hepatitis C. AB - BACKGROUND/AIMS: Liver negative-strand hepatitis C virus (HCV) RNA is the most direct indicator of active viral replication but has only been examined in a few semiquantitative studies. METHODS: Positive- and negative-strand HCV RNA in the right (R) and left (L) liver lobes was quantified by rTth-based strand-specific real-time polymerase chain reaction for 48 chronic hepatitis C patients. RESULTS: Close correlations between lobes were seen for positive- and negative-strand amounts (r = 0.950; P < 0.001 and r = 0.920; P < 0.001, respectively). The ratio of negative to positive strands (median, 0.14 for R and 0.13 for L) varied by 2 log directly in relation to HCV replication assessed by liver negative strands but had no relation to liver positive strands and circulating HCV. Only negative strand quantitation was inversely correlated with age (r = -0.322; P = 0.026 for R and r = -0.340; P = 0.018 for L), while liver tissues with hepatitis B virus DNA contained larger amounts of each strand. In 27 patients treated with enhanced interferon monotherapy, the amounts of liver negative strands (<4 log copies/100 ng RNA) were the only independent predictor of a sustained virologic response. CONCLUSIONS: Negative-strand quantitation is uniform in the liver and bears distinct relevance to the disease. PMID- 16343681 TI - HBV virological assessment. AB - Management of hepatitis B virus (HBV) infected patients involves serological diagnosis, quantitation of HBV-DNA and measurement of HBV drug resistance. Different serological markers such as HBsAg, anti-HBs, anti-HBc (total and IgM), HBeAg and anti-HBe are assessed by immunoassays in order to define the infection status. The emergence of surface mutants however is a continuous challenge to design more effective immunoassays. Commercially available quantitative HBV-DNA assays with increased sensitivity and wider linear range give a more accurate estimate of viral replication and contribute decisively in the initiation and the monitoring of the response to HBV therapy. Genotypic drug resistance assays are important diagnostic tools, since the administration of nucleoside/nucleotide analogues to HBV infected patients leads to the development of drug resistance patterns very much dependent on the treatment regimen. Special issues have to be taken into consideration regarding HBV/HIV-1 co-infected patients, since concominant HIV and HBV replication results in higher rates of HBV replication. Current efforts are focused on the standardization of HBV-DNA assays (qualitative and quantitative), of HBV drug resistance assays as well as in the development of new assays and markers that will help in the prognosis and management of HBV infection (quantitative detection of pre-core mutants and HBV ccc-DNA assays). PMID- 16343682 TI - HCV virological assessment. AB - Virological assessment of HCV infection relies on a series of assays that are essential for diagnostic purposes and to adopt therapeutic decisions. Diagnostic assays can be classified in three groups: assays that are designed to detect specific antibodies to HCV; techniques that detect and quantify viral load; and tests aimed at determining the infecting HCV type. In general, the use of these assays is similar in HCV infected patients as in those co-infected with HCV and HIV. Co-infected patients, however, have some inherent characteristics (deficient immune status, high HCV-RNA concentrations, particular genotype distribution, low response to antiviral therapy) that make the interpretation of such assays slightly different. Regarding assays to detect antibodies to HCV, last generation tests have a similar sensitivity in mono-infected and co-infected individuals. HCV-RNA testing might be helpful in anti-HCV negative individuals with clinical or analytical suspicion of liver disease. Genotype determination in patients co infected with HCV and HIV should be performed by methods relying on sequence analysis, since serotyping assays have shown a lower sensitivity in co-infected cohorts. HCV-RNA concentration is a strong predictor of response to therapy. Due to the higher viral load of co-infected patients compared to HCV mono-infected individuals, those assays with a wide dynamic range seem more appropriate to monitor viral load during treatment. PMID- 16343683 TI - Assessment of efficacy of treatment in HCV: infection and disease. AB - The hepatitis C virus (HCV) lifecycle appears to be confined to the cytoplasm of hepatocytes. This has translated into the ability to produce sustained virological responses (SVRs) in a substantial proportion of HCV-infected people. Lower treatment responses and higher relapse rates in co-infection may be a consequence of HCV more frequently infecting difficult-to-access reservoirs such as peripheral blood mononuclear cells. However, the durability of sustained response appears to be minimally altered in co-infection. In light of these modest SVRs, and accelerated hepatic fibrosis progression among co-infected people, strategies to slow disease progression take on greater importance. In this context, histological response becomes an important endpoint in co infection. Multicentre randomized trials of PEG-IFN and ribavirin demonstrate that assessment of histological responses either at the conclusion of therapy or 24 weeks after completion of therapy identify a number of patients who experience improvements in histological activity index in the absence of virological clearance. The prognostic value of serum markers of fibrosis in co-infection has not been validated, but this represents a particularly important area for development in co-infection in light of the probable need for more intensive histological sampling. Eventually, it is anticipated that genotype-phenotype correlations will permit selection of optimized antiviral regimens once virus specific enzymatic inhibitors have been approved. PMID- 16343684 TI - Natural history and predictors of severity of chronic hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection. AB - Co-infection by the hepatitis C virus (HCV) is observed in up to 30% of HIV infected individuals. In studies conducted in the 'pre-HAART era', the late consequences of HCV-related chronic liver disease were overshadowed by extra hepatic causes of deaths, related to severe immune deficiency, and the impact of HCV infection on mortality of HIV-infected patients was low. While the development of HAART has resulted in a significant decrease in morbidity and mortality amongst HIV-infected patients, this clear benefit allowed the expression of liver-related complications associated with HCV chronic infection. The impact of HCV on HIV remains debated but HIV infection significantly modifies the natural history of HCV infection. HIV infection increases levels of HCV viraemia by 2- to 8-fold, resulting in a significant decrease in spontaneous recovery of acute hepatitis. HIV co-infection also worsens the histological course of HCV infection by increasing and accelerating the risk of cirrhosis or leading to rare but lethal fibrosing cholestatic hepatitis. Liver disease is now one of the leading causes of morbidity and mortality in co-infected patients, even if HAART and especially protease inhibitors, may decrease the severity of the liver disease and the liver-related mortality. Several non-exclusive pathogenic processes explain the increasing rate of liver complications associated with HCV-related liver disease. PMID- 16343685 TI - Health care utilization among persons who are unemployed or outside the labour force. AB - BACKGROUND: In Sweden, equity in health is a central aim of public health policy. To this end, the health care system is obligated to offer equal access to health care according to need. However, unemployment may hinder the fulfillment of this goal. The aim of the present study was to assess self-reported health care needs and service utilization with respect to employment status. METHODS: A questionnaire was sent to 4000 randomly chosen individuals 20-64 years of age living in different counties in Sweden (response rate 66.2%). Logistic regression analyses were carried out to estimate the influence of employment status, socio demographic variables and health indicators on the need for and use of health care services. RESULTS: In total, 42.2% (n=35) among the unemployed, 37.4% (n=55) among persons who were on long-term sick leave (LTSD), and 22.3% (n=467) of the employed persons, abstained from consulting a physician despite reporting a perceived need to do so. The results persisted after adjusting for socio demographic variables, social support and personal finances (unemployed: OR=1.91; LTSD: OR=1.62). The risk of foregoing care remained higher among the unemployed, but not the LTSD-group, after adjusting for long-standing illness (OR=1.94). The unemployed were more likely than the employed to perceive a need to seek care for psychological problems. The risk of abstaining from consulting a physician was related to symptoms of depression. CONCLUSIONS: Lack of employment may be related to unmet care needs, especially among unemployed who are experiencing psychological symptoms. To deal with the needs of the unemployed it may be useful to develop interventions within the health care system that focus more on psychological problems. PMID- 16343686 TI - Profiling of resource use variation among six diseases treated at 82 Japanese special functioning hospitals, based on administrative data. AB - BACKGROUND: Profiling treatment in Japanese hospitals has rarely been conducted systematically with an administrative database. The study aims to present descriptive statistics of medical profiling and to examine the sources of variation in resource used for six common diseases. METHODS: Administrative records for 266,677 patients were analyzed to examine variation in length of stay (LOS) and total charge (TC) by hierarchical multiple linear regression for cases of ischemic stroke, ischemic heart disease (IHD), great vessel disease (GVD), respiratory neoplasm, gastric neoplasm and colonic neoplasm. RESULTS: Average LOS and TC increased with disease severity and invasiveness of surgical procedure. The coefficient of determination of the full model was highest for LOS in IHD (0.432), and for TC that was highest in GVD (0.702). Among various variable sets examined, surgical procedures explained largest variance in resource use. CONCLUSION: With a standardized database derived from claims data, wide audience of stakeholders in Japanese healthcare will be able to access the profiling of practice or disease variation concerned. PMID- 16343687 TI - Dying at home or in an institution using death certificates to explore the factors associated with place of death. AB - INTRODUCTION: The knowledge of determinants of place of death is important for public health policy aimed at improving the quality of end-of-life care. METHODS: We investigated the influence of clinical, socio-demographic, residential and health care system factors on the place of death, using data from all 55,759 deaths in 2001 in Flanders (Belgium), gathered via official death certificates and data from anonymously linked health care statistics. A multivariate logistic regression was used to examine the associated factors (home versus hospital as dependent categories). RESULTS: Of all deaths in Flanders, 53.7% took place in hospital, 24.3% at home and 19.8% in a care home. The probability of home deaths varied by region, by rural or urban residence and by the hospital bed availability in the region and dying at home was less likely among those suffering from certain non-malignant chronic diseases, the less educated and those living alone. CONCLUSION: Although most people wish to die at home, most deaths in Flanders (Belgium) in 2001 did not take place there. The clinical, socio-demographic and residential factors found to be associated with the place of death could serve as focal points for a policy to facilitate dying in the place of choice, including at home. PMID- 16343688 TI - Financial incentives for disease management programmes and integrated care in German social health insurance. AB - As a result of recent health care reforms sickness funds and health care providers in German social health insurance face increased financial incentives for implementing disease management and integrated care. Sickness funds receive higher payments form the risk adjustment system if they set up certified disease management programmes and induce patients to enrol. If health care providers establish integrated care projects they are able to receive extra-budgetary funding. As a consequence, the number of certified disease management programmes and the number of integrated care contracts is increasing rapidly. However, contracts about disease management programmes between sickness funds and health care providers are highly standardized. The overall share of health care expenses spent on integrated care still is very low. Existing integrated care is mostly initiated by hospitals, is based on only one indication and is not fully integrated. However, opportunity to invest in integrated care may open up innovative processes, which generate considerable productivity gains. What is more, integrated care may serve as gateway for the introduction of more widespread selective contracting. PMID- 16343689 TI - Mobility of pharmaceuticals carbamazepine, diclofenac, ibuprofen, and propyphenazone in miscible-displacement experiments. AB - Many pharmaceuticals pass the unsaturated zone before reaching an aquifer. Therefore, laboratory sand column transport experiments were conducted to study the transport behavior of carbamazepine, diclofenac, ibuprofen, and propyphenazone under unsaturated conditions. The test water was artificial sewage effluent to simulate the infiltration of reused wastewater. The test water was spiked with the pharmaceutically active compounds and the tracer LiCl. Afterwards it was passed through laboratory sand columns, one experiment for each pharmaceutical. The physical and chemical parameters were recorded and general ions measured. Pharmaceuticals were measured using solid phase extraction, derivatization, and detection with GC-MS. The column experiments indicate a significant elimination of ibuprofen (54%), propyphenazone (55%), and diclofenac (35%), whereas carbamazepine was not eliminated. Retardation factors varied between 1.84 for carbamazepine, 2.51 for propyphenazone, 3.00 for ibuprofen, and 4.80 for diclofenac. These results show that mobility and elimination of diclofenac, ibuprofen, and propyphenazone is about in the same range as for experiments under saturated conditions whereas carbamazepine had a significantly lower sorption and elimination under unsaturated conditions. PMID- 16343690 TI - Ascorbic acid oxidase is dynamically regulated by light and oxygen. A tool for oxygen management in plants? AB - Ascorbic acid oxidase (AAO) is a plant blue-copper protein catalyzing dioxygen reduction to water using ascorbic acid as the electron donor. In spite of extensive molecular characterization the physiological role of AAO is still uncertain. Abundant mRNA, protein and activity of AAO were observed in illuminated leaves of Cucurbita pepo. AAO activity was found to be proportional to light intensity. The light effect was rapidly reversed in dark and activity remained low throughout the dark period. Activity was elicited in dark by increased oxygen concentration. AAO activity increased in the facultative CAM Kalanchoe blossfeldiana upon induction of the CAM cycle and decreased during germination of C. pepo and Zea mays under hypoxic conditions. These results strongly suggest that AAO activity could be part of a dynamic system for oxygen management in plants. PMID- 16343691 TI - Evaluation of measures to decrease intra-operative bacterial contamination in orthopaedic implant surgery. AB - The aim of this study was to evaluate whether behavioural and systemic measures will decrease intra-operative contamination during total hip or knee replacements. The influence of these measures on subsequent prolonged wound discharge, superficial surgical site infection and deep periprosthetic infection was also investigated during an 18-month follow-up period. Four swabs were taken from instruments at the beginning and end of the procedure for 207 procedures. Removed bone material (acetabulum and femur in case of the hip joint; femur and tibia in case of the knee joint) was also tested for contamination. Initially, 70 operations performed under original control conditions were included, after which the first behavioural measure was introduced (i.e. better use of the plenum). Cultures were taken during 67 operations using better use of the plenum (Group 1), followed by disciplinary measures and the installation of a new laminar flow system. Seventy operations were monitored after this second intervention (Group 2). The control group showed intra-operative contamination in 32.9% (23/70) of cases, Group 1 showed contamination in 34.3% (23/67) of cases and Group 2 showed contamination in 8.6% (6/70) of cases. Prolonged wound discharge and superficial surgical site infection decreased significantly in Group 2, as did the incidence of deep periprosthetic infection; however, the latter did not reach statistical significance. This study shows that the combination of systemic and behavioural changes in an operating room significantly decreases the incidence of intra operative bacterial contamination, subsequent prolonged wound discharge and superficial surgical site infection. After 18 months of follow-up, there was also a decrease in deep periprosthetic infection. PMID- 16343692 TI - The neuronal histamine H(1) and pro-opiomelanocortin-melanocortin 4 receptors: independent regulation of food intake and energy expenditure. AB - Hypothalamic neuronal histamine and its H(1) receptor (H(1)-R) form part of the leptin signaling pathway in the brain, and regulate body weight and adiposity by affecting food intake and energy expenditure. The pro-opiomelanocortin (POMC) melanocortin 4 receptor (MC4-R) is also important for leptin signaling. We investigated whether and how these two neuronal pathways interact in regulating energy metabolism. From studies of agouti yellow (A(y)/a) obese mice, a model of a defect in POMC-MC4-R signaling, we concluded that the histamine H(1)-R signaling pathway is independent of the POMC-MC4-R complex in regulating food intake, energy metabolism, and adiposity. PMID- 16343693 TI - Structure-activity relationship analysis of antioxidant ability and neuroprotective effect of gallic acid derivatives. AB - Gallic acid and its derivatives are a group of naturally occurring polyphenol antioxidants which have recently been shown to have potential healthy effects. In order to understand the relationship between the structures of gallic acid derivatives, their antioxidant activities, and neuroprotective effects, we examined their free radical scavenging effects in liposome and anti-apoptotic activities in human SH-SY5Y cell induced by 6-hydrodopamine autooxidation. It was found that these polyphenol antioxidants exhibited different hydrophobicity and could cross through the liposome membrane to react with 1,1-diphenyl-2-picryl hydrazyl (DPPH) free radical in a time and dose-dependent manner. At the same time, the structure-antioxidant activity relationship of gallic acid derivatives on scavenging DPPH free radical in the liposome was also analyzed based on theoretical investigations. Analysis of cell apoptosis, intracellular GSH levels, production of ROS and the influx of Ca(2+) indicated that the protective effects of gallic acid derivatives in cell systems under oxidative stress depend on both their antioxidant capacities and hydrophobicity. However, the neuroprotective effects of gallic acid derivatives seem to depend more on their molecular polarities rather than antioxidant activities in the human SH-SY5Y cell line. In conclusion, these results reveal that compounds with high antioxidant activity and appropriate hydrophobicity are generally more effective in preventing the injury of oxidative stress in neurodegenerative diseases. PMID- 16343694 TI - Anti-Parkinsonian agents have anti-amyloidogenic activity for Alzheimer's beta amyloid fibrils in vitro. AB - Inhibition of the accumulation of amyloid beta-peptide (Abeta) and the formation of beta-amyloid fibrils (fAbeta) from Abeta, as well as the destabilization of preformed fAbeta in the central nervous system would be attractive therapeutic targets for the treatment of Alzheimer's disease (AD). Many studies have demonstrated that oxidative damage plays a central role in AD pathogenesis, as well as Parkinson disease (PD). Among the antioxidant strategies proposed, increasing evidence points to the possibility of achieving neuroprotection by dopamine agonists, as well as monoamine oxidase B (MAO-B) inhibitors. Actually, the beneficial effect of selegiline, a MAO-B inhibitor, in AD has been noted in several clinical studies. On the reverse, antimuscarinic agents have been reported to accelerate beta-amyloidosis and senile plaque formation in PD. Using fluorescence spectroscopic analysis with thioflavin T and electron microscopic studies, we examined the effects of anti-Parkinsonian agents, dopamine, levodopa, pergolide, bromocriptine, selegiline, and trihexyphenidyl on the formation, extension, and destabilization of fAbeta(1-40) and fAbeta(1-42) at pH 7.5 at 37 degrees C in vitro. The anti-Parkinsonian agents other than trihexyphenidyl dose dependently inhibited fAbeta formation from Abeta(1-40) and Abeta(1-42), as well as their extension. Moreover, these agents dose-dependently destabilized preformed fAbetas. The overall activity of the molecules examined was in the order of: dopamine>selegiline>levodopa=pergolide>bromocriptine. Although the exact mechanism of the anti-amyloidogenic activity of these agents is unclear, these and other structurally related compounds could be key molecules for the development of therapeutics for AD and other conformational diseases. PMID- 16343695 TI - Tetrahydrobiopterin causes mitochondrial dysfunction in dopaminergic cells: implications for Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder associated with a selective loss of dopaminergic neurons in the substantia nigra. While the underlying cause of PD is not clearly understood, oxidative stress and mitochondrial dysfunction are thought to play a role. We have previously suggested tetrahydrobiopterin (BH4), an obligatory cofactor for the dopamine synthesis enzyme tyrosine hydroxylase and present selectively in monoaminergic neurons in the brain, as an endogenous molecule that contributes to the dopaminergic neurodegeneration. In the present study, we show that BH4 leads to inhibition of activities of complexes I and IV of the electron transport chain (ETC) and reduction of mitochondrial membrane potential. BH4 appears to be different from rotenone and MPP(+), the synthetic compounds used to generate Parkinson models, in its effect on complex IV. BH4 also induces the release of mitochondrial cytochrome c. Pretreatment with the sulfhydryl antioxidant N acetylcysteine or the quinone reductase inducer dimethyl fumarate prevents the ETC inhibition and cytochrome c release following BH4 exposure, suggesting the involvement of quinone products. Together with our previous observation that BH4 leads to generation of oxidative stress and selective dopaminergic neurodegeneration both in vitro and in vivo via inducing apoptosis, the mitochondrial involvement in BH4 toxicity further suggests possible relevance of this endogenous molecule to pathogenesis of PD. PMID- 16343696 TI - Tau protein abnormalities associated with the progression of alzheimer disease type dementia. AB - The degree to which neurofibrillary tangles (NFT), the hallmark lesions of Alzheimer disease (AD), contribute to the development of the cognitive symptoms of AD has been debated. NFTs are comprised of abnormally phosphorylated and conformationally altered tau proteins. Conformational changes in tau have been proposed to be among the earliest neurobiological changes in AD. This study examined whether conformational changes detected by antibodies MC1 and TG3 represent early abnormalities in the disease process by assessing their presence at different stages of dementia in multiple brain regions. Postmortem specimens from several neocortical regions were examined for conformational changes in tau by ELISA in subjects [n=81] who died at different stages of cognitive impairment. Concentrations of conformationally altered tau increased with increasing dementia severity and the levels of MC1 immunoreactivity increased in the frontal cortex of mildly demented subjects before the appearance of NFT bearing neurons, suggesting that conformational alterations in tau occur early in the course of AD and its cognitive symptoms and may precede histologically identified NFTs. PMID- 16343697 TI - The Val66Met polymorphism of the brain-derived neurotrophic-factor gene is associated with geriatric depression. AB - Brain-derived neurotrophic-factor (BDNF), the most abundant of the neurotrophins in the brain, has been implicated in both major depression and cognitive function. This study examines the association between the BDNF-gene Val66Met polymorphism and depression susceptibility and severity, age-of-onset, cognitive function and suicidal attempt history in an elderly Chinese sample population. We genotyped the BDNF-gene Val66Met polymorphism in 110 elderly inpatients diagnosed with major depression and 171 age- and sex-similar control subjects. All patients were assessed with the Hamilton Rating Scale for Depression (HAM-D) for depression severity and the Mini-Mental Status Examination (MMSE) for cognitive function after admission. Suicide attempt history and age-of-onset of depression were evaluated by interview and medical record. The BDNF Val66Met genotype distribution was significantly different between depressed patients and control subjects (P=0.003) and there was a significant excess of Met allele in the depressed patients compared to the control group (P=0.001). The BDNF polymorphism did not affect age-of-onset, depression severity, cognitive function or suicidal attempt history. The results suggest that the BDNF Val66Met polymorphism is a relevant risk factor for geriatric depression. PMID- 16343698 TI - Evaluating the efficacy of an integrated curriculum on adolescent health and development for pre-service nursing education in Hong Kong. AB - BACKGROUND: An integrated adolescent curriculum on health and development (ADH) was implemented in a pre-service nursing programme in a university. AIM: This study examined the efficacy of an ADH curricular framework in improving the competency variables of student nurses in delivering ADH services. METHOD: The design of the study was quasi-experimental with the systematically designed ADH training curriculum as an intervention. Pre- and post-tests incorporating an experimental group and a control group for a sample of 101 student nurses were used. FINDINGS: Reports from 50 student nurses in the experimental group indicated that there was a significant increase from the pre-test to the post test phase in the total score for the variables in the ADH Competency Checklist (Z=-5.71, p<0.001) and its four subscales: the professional development subscale (Z=-5.37, p<0.001), the psychosocial and physical well-being subscale (Z=-5.66, p<0.001), the health behaviours and lifestyles subscale (Z=-5.07, p<0.001), and the identity and reproductive health subscale (Z=-4.86, p<0.001). Significant changes were detected in the ADH competency variables for the control group in the post-test phase. CONCLUSION: The findings reveal that the systematic integration of ADH in the nursing curriculum had the positive impact of increasing the competency of student nurses for the examined variables. PMID- 16343699 TI - Ganglioside GM1-binding peptides as adjuvants of antigens inoculated by the intranasal route. AB - Forty-five GM1-binding peptides were identified using phage-displayed peptides libraries of random peptides. Most have a motif containing a hydrophobic amino acid followed by a serine (S). Based on a GM1-binding assays, two of these GM1 binding peptides (named 15 and 40) were chosen to investigate its immunostimulatory properties when chemically coupled to antigens. Mice intra nasally (i.n.) vaccinated with some of these complexes developed a better local and systemic antibody response than mice i.n. vaccinated with the respective uncoupled antigens. The efficiency of the complex GM1-binding peptide-antigen strongly depends on the composition and structure of both of the components of the complex. PMID- 16343700 TI - Antibody induction after combined application of an adjuvanted recombinant FeLV vaccine and a multivalent modified live virus vaccine with a chlamydial component. AB - The compatibility, safety and interaction on antibody induction of a combined vaccine application were assessed. Specific pathogen-free cats were vaccinated with either a modified live virus vaccine containing feline calici- (FCV), herpes (FHV-1), parvovirus (FPV) and Chlamydophila felis (C. felis), an adjuvanted recombinant feline leukaemia virus (FeLV) vaccine or both vaccines in one syringe. After combined application, FeLV ELISA antibody titres were unaltered, However antibody production based on indirect immunofluorescence assay was remarkably enhanced for FCV and was at selected time points also enhanced for FHV 1 and C. felis but diminished for FPV. The use of these vaccines in combination was safe and will simplify vaccination schedules in veterinary practice. PMID- 16343701 TI - Vaccination of chicken embryos with escape mutants of La Sota Newcastle disease virus induces a protective immune response. AB - To reduce the embryonic pathogenicity of Newcastle disease virus (NDV), escape mutants of the La Sota strain were produced with selected monoclonal antibodies. Immunoselection resulted in the elimination of an epitope by single amino acid substitution (F and HN molecule) or in a conformational change (HN molecule). The embryonic pathogenicity of these escape mutants was reduced and their dose was optimised for in ovo vaccination. Because antibody responses and protection of in ovo vaccinated chicks were similar to controls vaccinated at hatch with the La Sota strain, immunoselection appears a valuable technique to produce attenuated NDV strains, which are candidate in ovo vaccines. PMID- 16343702 TI - The New World primate, Aotus nancymae, as a model for examining the immunogenicity of a prototype enterotoxigenic Escherichia coli subunit vaccine. AB - The colonization factors (CF) of enterotoxigenic Escherichia coli (ETEC) are being targeted for inclusion in a multi-subunit ETEC vaccine. This study was designed to examine the preclinical safety and immunogenicity of CF CS6, encapsulated in a biodegradable poly(DL-lactide-co-glycolide) (meCS6), and administered in the presence or absence of a mutated heat-labile enterotoxin, LT(R192G), in the non-human primate, Aotus nancymae. A. nancymae were inoculated intranasally (IN) with meCS6 (200 microg; positive control), or intragastrically (IG) with meCS6 (200 or 1000 microg) with or without 2 microg LT(R192G) in three doses given at 2-week intervals. In a second experiment, A. nancymae were inoculated IG with 950 microg of meCS6 with or without 2 microg LT(R192G) in four doses given every 48 h. Blood was collected to assess anti-CS6 and -LT serum immunoglobulin G (IgG) and IgA responses and safety variables (complete blood count and chemistry). Safety parameters were unchanged from baseline following all vaccinations. In Experiment 1, a dose-related serologic response to CS6 was observed; 78.6 and 57.1% of monkeys given 1000 microg meCS6 (n = 14) had a serum IgG and IgA response, respectively, compared to only 28.6% of monkeys given 200 microg meCS6 (n = 14) with a serum IgG and IgA response. No significant effect on the number of responders or the magnitude of responses was observed with the addition of LT(R192G). The three-dose, 2-week regimen with 1000 microg meCS6 was more effective at eliciting an immune response than the four-dose, 48-h regimen with 950 microg meCS6. Results from this study indicate that A. nancymae provide a useful ETEC preclinical safety and immunogenicity model. PMID- 16343703 TI - Characterization of live influenza vaccine donor strain derived from cold adaptation of X-31 virus. AB - A human influenza A virus X-31 (high-yielding strain) was cold-adapted for possible future use as live attenuated vaccine. Mutant influenza viruses were selected during successive serial passage in embryonated hens' eggs at progressively lower sub-optimal temperature (30, 27 degrees C followed by 24 degrees C). The cold-passaged mutant exhibited both temperature-sensitivity (ts) and cold-adapted (ca) phenotypes. The pathogenicity and immunogenicity of X-31 ca virus were studied in mice following intranasal inoculation. The mice did not show clinical signs even at high titer infection. Immunization of mice with X-31 ca virus elicited high titers of neutralizing antibody and provided complete protection against homologous and heterologous virus challenges. To assess the genetic stability, the X-31 ca virus was passaged at 37 degrees C in MDCK cells or inoculated into mice. Revertant virus was not found in the lungs of any of the mice and the supernatants of the MDCK culture. We conclude that the X-31 ca candidate vaccine virus exhibits the desired level of attenuation, immunogenicity, and protective efficacy required for live attenuated vaccine and merits further evaluation at clinical level. PMID- 16343704 TI - Immunisation with recombinant BCG expressing the cottontail rabbit papillomavirus (CRPV) L1 gene provides protection from CRPV challenge. AB - Recombinant Bacille Calmette-Guerin (rBCG) could potentially be the vaccine vehicle of choice to deliver foreign antigens from multiple pathogens. In this study we have used the cottontail rabbit papillomavirus (CRPV) rabbit model to provide a "proof of concept" that immunisation with rBCG expressing the CRPV major capsid protein, L1 (rBCG/CRPVL1), will protect outbred New Zealand White rabbits against CRPV challenge. Rabbits immunised with rBCG/CRPVL1 (10(7) cfu/ml) were protected 5 weeks post-CRPV challenge. Rabbits immunised with rBCG/CRPVL1 (10(5) cfu/ml) had papillomas, which were smaller and took longer to appear than the control rabbits. None of the negative control rabbits vaccinated with rBCG expressing an irrelevant gene or PBS were protected from CRPV challenge. Sera from rabbits immunised with rBCG/CRPVL1 (10(7) cfu/ml) were able to neutralise 54.5% of CRPV at serum dilutions of 1:200. These results provide evidence that BCG could potentially be used as a vaccine delivery vehicle for human papillomavirus proteins as a possible prophylactic vaccine. PMID- 16343705 TI - Resistance of the cholera vaccine candidate IEM108 against CTXPhi infection. AB - The cholera toxin (CT) genes ctxAB are carried on a lysogenic phage of Vibrio cholerae, CTXPhi, which can transfer ctxAB between toxigenic and nontoxigenic strains of bacteria. This transfer may pose a problem when live oral cholera vaccine is given to people in epidemic areas, because the toxin genes can be reacquired by the vaccine strains. To address this problem, we have constructed a live vaccine candidate, IEM108, which carries an El Tor-derived rstR gene. This gene encodes a repressor and can render bacterial resistance to CTXPhi infection. In this study, we evaluated the resistance of IEM108 against CTXPhi infection by using a CTXPhi marked for chloramphenicol (CAF) resistance and an in vivo model. We found that the cloned rstR gene rendered IEM108 immune to infection with the marked CTXPhi. In addition, the infection rate of IEM108 was even lower than that of the native CTXPhi-positive strain. These results suggest that the vaccine candidate IEM108 is resistant to infection by CTXPhi. PMID- 16343706 TI - The Brazilian National Intercomparison Program (PNI/IRD/CNEN): evaluation of 15 years of data. AB - The Instituto de Radioprotecao e Dosimetria has been coordinating the National Intercomparison Program (PNI) for 15 years, from 1991 to 2005. The objective is to evaluate the analytical performance of the laboratories in low-level activity concentration radionuclide assays in environmental samples. This work presents an evaluation of PNI data from 42 intercomparison runs, which distributed to 22 Brazilian laboratories 2511 samples evaluating 5768 radionuclide assays involving 32 radionuclides in the period from 1991 to 2004. The laboratory performance was evaluated using the Normalized Standard Deviation used by the U.S. EPA. For comparison aims, the Normalized Deviation, used by BIPM was also applied. Laboratory performance were grouped and evaluated in three periods of 5 years each. The first period shows, an average value of good performance of 71.2%, the second shows an average of 78.6% and the last an average of 82.3%. Performance for each kind of radionuclide is also presented. PMID- 16343707 TI - Anxiety and self-consciousness in patients with facial lacerations one week and six months later. AB - We explored the psychological impact of disfigurement resulting from facial lacerations in patients who presented at an accident and emergency department in the UK. Patients participated in relevant sections of the Derriford Appearance Scale and State Trait Anxiety Inventory at one week and six months after injury. In all patients self-consciousness and anxiety scores were higher than in the general population. The extent of the scar, the patient's living arrangements, and the cause of the injury were significantly related to self-consciousness and anxiety. General self-consciousness improved at six months but not social self consciousness and anxiety. These injuries may have a considerable impact on the lives of the affected people. PMID- 16343708 TI - World Health Organization/International Confederation of Midwives Collaboration: pathways to healthy nations. AB - In this paper, the collaboration between the World Health Organization (WHO) and the International Confederation of Midwives (ICM) to reduce maternal mortality and morbidity, and baby mortality and morbidity, is explicated. PMID- 16343709 TI - Brachialis muscle transfer to the forearm muscles in obstetric brachial plexus palsy. AB - Tendon transfers are frequently needed to improve hand function in obstetric brachial plexus injuries. The reconstruction cannot always be achieved using local donor transfers in the forearm as these are not always available. In such cases, we propose the use of the brachialis muscle as a useful donor for transfer. Five adolescents with obstetric brachial plexus palsy were operated on to reconstruct wrist extension and/or pronation using the brachialis muscle transfer to the pronator teres (n=1), extensor carpi radialis brevis (n=1) and extensor carpi radialis longus (n=3). Twelve months after surgery, average active motion recovery was 20 degrees for wrist extension and 14 degrees for pronation. Active and passive range of motion was similar. PMID- 16343711 TI - Re: a randomised comparison of locking and non-locking palmar plating for unstable Colles' fractures in the elderly, Koshimune et al., Journal of Hand Surgery, 2005, 30B: 499-503. PMID- 16343712 TI - Re: the role of antibiotic prophylaxis in clean incised hand injuries: a prospective randomized placebo controlled double blind trial, Whittaker JP, Nancarrow JD, Sterne GD, Journal of Hand Surgery, 2005, 30B: 162-167. PMID- 16343713 TI - Validation of a new protocol for computer-assisted evaluation of kinematics of double-bundle ACL reconstruction. AB - BACKGROUND: Computer-assisted surgery is useful to increase the precision of anterior cruciate ligament (ACL) surgical procedure, but could be even more important in evaluating the global performance of reconstructed ACL. This paper describes a new protocol for an accurate and extensive computer-assisted evaluation of single- and double-bundle reconstructions of ACL. METHODS: The protocol consists of the acquisition of the leg axes, ACL and graft insertions by a navigation system, and tracking of the knee motion during the classical kinematic test of knee stability. These data are elaborated by computer software in order to compute graft biomechanical behaviour and the knee kinematics and estimate the performance of the intervention. FINDINGS: The proposed protocol was validated on three cadaver knees. It resulted minimally invasive, effective to describe graft kinematic performance and able to provide a 3D reliable description of the reconstructed knee. INTERPRETATION: The protocol is an extension of the present evaluation of computer-assisted packages and includes additional kinematic tests and computations. The scientist-reader can find important details on tested computations to implement a similar computer-assisted procedure for new applications in knee surgery, while the surgeon can find in this procedure a means to improve the evaluation of ACL reconstruction and identify the residual laxity. PMID- 16343714 TI - Field responses of Prunus serotina and Asclepias syriaca to ozone around southern Lake Michigan. AB - Higher ozone concentrations east of southern Lake Michigan compared to west of the lake were used to test hypotheses about injury and growth effects on two plant species. We measured approximately 1000 black cherry trees and over 3000 milkweed stems from 1999 to 2001 for this purpose. Black cherry branch elongation and milkweed growth and pod formation were significantly higher west of Lake Michigan while ozone injury was greater east of Lake Michigan. Using classification and regression tree (CART) analyses we determined that departures from normal precipitation, soil nitrogen and ozone exposure/peak hourly concentrations were the most important variables affecting cherry branch elongation, and milkweed stem height and pod formation. The effects of ozone were not consistently comparable with the effects of soil nutrients, weather, insect or disease injury, and depended on species. Ozone SUM06 exposures greater than 13 ppm-h decreased cherry branch elongation 18%; peak 1-h exposures greater than 93 ppb reduced milkweed stem height 13%; and peak 1-h concentrations greater than 98 ppb reduced pod formation 11% in milkweed. PMID- 16343715 TI - Calibration of the Chemcatcher passive sampler for the monitoring of priority organic pollutants in water. AB - An integrative passive sampler consisting of a C18 Empore disk receiving phase saturated with n-octanol and fitted with low-density polyethylene diffusion membrane was calibrated for the measurement of time-weighted average concentrations of hydrophobic micropollutants, including polyaromatic hydrocarbons and organochlorine pesticides, in water. The effect of temperature and water turbulence on kinetic and thermodynamic parameters characterising the exchange of analytes between the sampler and water was studied in a flow-through system under controlled conditions. It was found that the absorption of test analytes from water to the sampler is related to their desorption to water. This allows for the in situ calibration of the uptake of pollutants using offload kinetics of performance reference compounds. The sampling kinetics are dependent on temperature, and for most of the tested analytes also on the flow velocity. Sampler-water partition coefficients did not significantly change with temperature. PMID- 16343716 TI - Benthic foraminifera and heavy metals distribution: a case study from the Naples Harbour (Tyrrhenian Sea, Southern Italy). AB - The analysis of 90 surficial sediments from three docks of the Naples Harbour (Levante, Granili, and Diaz) permits to compare the distribution modes of heavy metals with grain sizes, total organic carbon content (TOC) and distribution patterns of benthic foraminifera. Foraminiferal density and species richness decrease with the increasing toxic elements concentrations from the Levante to the Diaz dock. Median concentrations of Ni, Pb, Zn, and Hg (medians of 21.43 mg/kg, 270.24 mg/kg, 489.65 mg/kg, and 1.18 mg/kg, respectively) were reported for the Diaz dock where foraminifera are absent, thus suggesting a possible impact of toxic elements on the benthic ecosystem balance. Compared to the unpolluted marine sediments of the Granili dock, the Levante area shows higher heavy metals levels and a quasi-oligotypic benthic assemblage. This is dominated by the tolerant species Ammonia tepida that may be used as bio-indicator of pollution of anthropised marine sediments. PMID- 16343717 TI - Brachidontes variabilis and Patella sp. as quantitative biological indicators for cadmium, lead and mercury in the Lebanese coastal waters. AB - The mussel, Brachidontes variabilis, and the limpet, Patella sp., were used as indicators to monitor cadmium, lead and mercury concentrations along the Lebanese coast. Studies were carried out in order to define the best strategy for assessing and minimizing the effects of size and physiological condition on the metal contents of the molluscs, and corrective models were constructed. Metal concentrations in surface water were measured to estimate bioconcentration factors (BCFs). The BCFs varied from 8.3 x 10(3) to 3.4 x 10(4), from 7.5 x 10(3) to 8.0 x 10(3) and from 3.0 x 10(4) to 3.2 x10(4), for Cd, Pb and Hg, respectively. For limpets, BCFs varied from 1.7 x 10(4) to 7.4 x 10(4) for Cd, from 2.5 x 10(3) to 6 x 10(3) for Pb and remained fairly constant at around 10(4) for Hg. The highest BCFs were associated with lowest contamination levels. The results of the geographical survey exhibited a similar large-scale spatial pattern for the two species and followed the metal concentration distributions measured in the waters. PMID- 16343718 TI - Efficient retrieval of vegetation leaf area index and canopy clumping factor from satellite data to support pollutant deposition assessments. AB - Canopy leaf area index (LAI) is an important structural parameter of the vegetation controlling pollutant uptake by terrestrial ecosystems. This paper presents a computationally efficient algorithm for retrieval of vegetation LAI and canopy clumping factor from satellite data using observed Simple Ratios (SR) of near-infrared to red reflectance. The method employs numerical inversion of a physics-based analytical canopy radiative transfer model that simulates the bi directional reflectance distribution function (BRDF). The algorithm is independent of ecosystem type. The method is applied to 1-km resolution AVHRR satellite images to retrieve a geo-referenced data set of monthly LAI values for the conterminous USA. Satellite-based LAI estimates are compared against independent ground LAI measurements over a range of ecosystem types. Verification results suggest that the new algorithm represents a viable approach to LAI retrieval at continental scale, and can facilitate spatially explicit studies of regional pollutant deposition and trace gas exchange. PMID- 16343719 TI - Atmospheric polycyclic aromatic hydrocarbons (PAHs) in Asia: a review from 1999 to 2004. AB - Polycyclic aromatic hydrocarbons (PAHs) are present in both gaseous and particulate phases. These compounds are considered to be atmospheric contaminants and are human carcinogens. Many studies have monitored atmospheric particulate and gaseous phases of PAH in Asia over the past 5 years. This work compares and discusses different sample collection, pretreatment and analytical methods. The main PAH sources are traffic exhausts (AcPy, FL, Flu, PA, Pyr, CHR, BeP) and industrial emissions (BaP, BaA, PER, BeP, COR, CYC). PAH concentrations are highest in areas of traffic, followed by the urban sites, and lowest in rural sites. Meteorological conditions, such as temperature, wind speed and humidity, strongly affect PAH concentrations at all sampling sites. This work elucidates the characteristics, sources and distribution, and the healthy impacts of atmospheric PAH species in Asia. PMID- 16343720 TI - A critical review of theories to explain violent relationship termination: implications for research and intervention. AB - The following review represents an update and extension to an influential article (see [Strube, M.J. (1988). The decision to leave an abusive relationship: Empirical evidence and theoretical issues. Psychological Bulletin, 104, 236 250.]), which evaluated empirical research and suggested potential theories to explain victimized women's decisions to terminate violent relationships. In contrast to the original review, this paper provides information on the importance of and means by which theory should be evaluated and critically determines which theoretical approach(es) might be most productive based on theoretical and atheoretical evidence. In addition, this paper discusses strengths and weaknesses of each approach, in light of certain criteria deemed to be important for the evaluation of theory (e.g., comprehensiveness, parsimony, etc.). Furthermore, this paper discusses current controversies regarding these issues, ramifications of differing theoretical approaches, and their potential impact on the field. Based on this analysis, it is determined that general approaches (e.g., reasoned action/planned behavior, investment model) may be better for understanding this complex and multifaceted decision. Suggestions for future theoretical and intervention research are discussed. PMID- 16343721 TI - The stratification of social capital and its consequences for self-rated health in Taganrog, Russia. AB - Russian public health and its social determinants have been the theme of several recent studies. In one of these, Rose [(2000). How much does social capital add to individual health? A survey study of Russians. Social Science & Medicine, 51(9), 1421-1435] puts forward a composite model as a way of getting away from two traditions: one that postulates that social capital influences health independently of human capital attributes (education, social class, income, etc.) and one that postulates that human capital is the main determinant of health, while social capital is more or less irrelevant. In this study, we investigate the composite model, conceptualising social capital as a type of capital, on the basis of Bourdieu. By doing this, not only do the relations between social capital and other types of capital become relevant, but also whether the effect of social capital on health differs depending on the possession of other types of capital. We used the Taganrog survey of 1998 which used structured interviews with the family members of 1,009 households and the response rate was 81%. We found that social capital is stratified by education, and also that its effect on health varies depending on the volume of educational capital possessed. It also seems to be extremely important to specify different types of social capital, in order to get a better overview of possible mechanisms by means of which different types of capital might affect health. PMID- 16343722 TI - Sharing decisions in consultations involving anti-psychotic medication: a qualitative study of psychiatrists' experiences. AB - In psychiatry, and in treating people with a diagnosis of schizophrenia in particular, there are obstacles to achieving concordant, shared decision making and in building a co-operative therapeutic alliance where mutual honesty is the norm. Studies of people with a diagnosis of schizophrenia have revealed critical views of medical authority, particularly over the issue of enforced compliance with antipsychotic medication. Psychiatrists are known to place particular value on such medication. This qualitative study reports the views of 21 general adult psychiatrists working in UK about their experiences of consultations involving discussion of antipsychotic medication. Interviewees reported a general commitment to achieving concordant relationships with patients and described a number of strategies they used to promote this. In this respect, their self perception differs from the picture of authoritarian practice painted by critics of psychiatry, and by some studies reporting patients' views. Interviewees also described obstacles to achieving concordance, including adverse judgements of patients' competence and honesty about their medication use. Explaining the adverse effects of medication was perceived to discourage some patients from accepting this treatment. Moments of strategic dishonesty were reported. Psychiatrists perceived that trust could be damaged by episodes of coercion, or by patients' perception of coercive powers. We conclude that a self-perception of patient-centredness may not preclude psychiatrists from fulfilling a social control function. PMID- 16343723 TI - Why do women consent to surgery, even when they do not want to? An interactionist and Bourdieusian analysis. AB - The 'informed consent' process has been placed at the centre of bioethical and policy discourses about how the autonomy and rights of patients can best be protected. Although there has been critical analysis of how the process functions in relation to participation in research and particular ethical 'dilemmas', there has been little examination of the routine business of consenting to medical procedures. Evidence is now beginning to emerge that people may consent to surgery even when reluctant to do so. In this paper, we develop an analysis informed by Bourdieusian and interactionist social theory of the accounts of 25 British women who consented to surgery in obstetrics and gynaecology. Of these, nine were ambivalent or opposed to having an operation. When faced with a consent form, women's accounts suggest that they rarely do anything other than obey professionals' requests for a signature. Women's capacity to act is reduced as they become enmeshed in the hospital structure of tacit, socially imposed rules of conduct. However, the interactionist account of power operating through the social rules of particular situated encounters, and the sanctions associated with rule-breaking, may not provide a sufficiently powerful explanation for why women submit to surgery they are opposed or ambivalent towards. Bourdieu's concepts of habitus, capital and symbolic power/violence offer a potentially more elaborated account, by showing how the practical logic that women apply in the field of surgery confers a 'sense of place' relative to professionals. Women experience deficits in capital, intensified by their physical vulnerability in critical situations, that severely constrain their ability to exercise choice. This work demonstrates the weakness of the consent process as a safeguard of autonomy. Far from reinforcing autonomy, the process may reinforce rather than disrupt passivity, but more generally our findings raise the question of the extent to which autonomy is an illusory goal. PMID- 16343724 TI - The work to make telemedicine work: a social and articulative view. AB - This article contends that the take up of telemedicine results inevitably in the reconfiguration of the existing work practices and socio-material relationships. This new way of working triggers a variety of shifts in coordination mechanisms, work processes and power relationships in the health care sector. The paper, which is based on the findings of a research project conducted in Northern Italy, addresses three critical issues of telemedicine: the conflict between the scripts embodied in telemedicine technologies and the daily work practices of heath care professionals; the tendency of telemedicine to produce a delegation of medical tasks to non-medical personnel (and to artifacts); and the tendency of telemedicine to modify the existing geography within the health care environment. The paper contends that telemedicine presupposes and entails some significant changes in work processes which affect both the material conditions of the expertise which is supposed to be distributed, and the relationships between health care professionals and their practices. PMID- 16343725 TI - Alpha-tocopherol protects against the renal damage caused by potassium dichromate. AB - Exposure to hexavalent chromium (Cr(6+)) causes mutagenic, carcinogenic, and toxic effects, some of which have been associated with its oxidative capacity. In the kidney, Cr(6+) has been claimed to provoke necrosis of the proximal tubular cells. Our aim was to assess the functional involvement of the different segments that form the nephron in a model of acute renal failure caused by potassium dichromate and the participation of oxidative damage in this process. We also studied the possible protective effect of alpha-tocopherol (alpha-TOC) against renal damage. Wistar female rats 200g body weight (bw) received potassium dichromate (15mg/kg, sc, single dose). Lipid peroxidation and renal function were evaluated on days 0, 1, 2, 3, 7, 10, and 14. A second group received alpha-TOC (125mg/kg, by gavage) 5 days before and during dichromate exposure (same dose as for the first group), and was monitored at 0, 2, and 7 days of exposure. Creatinine clearance, glucose and sodium fractional excretions, p-aminohippurate uptake, free-water and osmolal clearances were also measured. Thiobarbituric acid reactive substances were quantified in renal cortex. The results revealed altered proximal tubule function, decreased glomerular filtration, and distal segment dysfunction, accompanied by oxidative damage 48h after exposure to dichromate. In the alpha-TOC-treated group proximal reabsorptive and secretory functions were preserved, suggesting that oxidative damage is a participating mechanism in dichromate toxicity on these functions. In contrast alpha-TOC did not prevent glomerular or distal dysfunction, indicating selectivity of the protection afforded by this compound on the toxicity of dichromate, at the several components of the nephron. PMID- 16343726 TI - Statins induce calcium-dependent mitochondrial permeability transition. AB - Statins (3-hydroxy-3-methylglutaryl-CoA reductase inhibitors) are used in the treatment of hypercholesterolemic patients to reduce risk of cardiovascular diseases because of their cholesterol lowering action. Other lipid independent protective actions of statins have been reported. However, some adverse side effects have, also, been described. We report, here, that liver mitochondria isolated from hypercholesterolemic LDL receptor knockout mice treated during 15 days with therapeutic doses (100 mg/kg, p.o.) of lovastatin presented a higher susceptibility to develop membrane permeability transition (MPT). In experiments in vitro, lovastatin-induced MPT in a dose-dependent manner (10-80 microM) by a mechanism sensitive to cyclosporin A (cyclophilin sequestrant), dithiothreitol (reducing agent), adenine nucleotide carrier inhibitor (ADP), catalase (H2O2 reductant) and EGTA (calcium chelator). In agreement with the inhibition of the mitochondrial swelling by dithiothreitol, lovastatin, also, decreased the content of total mitochondrial membrane protein thiol groups. Simvastatin had similar effects on mitochondria; however, pravastatin, a hydrophilic statin, had a weaker effect in inducing MPT. In conclusion, statins can act directly on mitochondria either in vivo or in vitro inducing permeability transition, which is a process involved in cell death. PMID- 16343727 TI - Age-dependent pharmacokinetic and pharmacodynamic response in preweanling rats following oral exposure to the organophosphorus insecticide chlorpyrifos. AB - Previous studies have indicated that juvenile rats are more susceptible than adults to the acute toxicity from exposure to the organophosphorus insecticide chlorpyrifos (CPF) and age-dependent differences in metabolism and sensitivity to cholinesterase (ChE) inhibition may be responsible. Metabolism involves CYP450 activation and detoxification of CPF to CPF-oxon and 3,5,6-trichloro-2-pyridinol (TCP), as well as cholinesterase (acetyl- and butyrylcholinesterase), carboxylesterase (CaE), and A-esterase (PON-1) detoxification of CPF-oxon to TCP. The pharmacokinetics of CPF, TCP, and the extent of blood (plasma/RBC), and brain ChE inhibition in rats were determined on postnatal days (PND)-5, -12, and -17 following oral gavage administration of 1 and 10mg CPF/kg of body weight. As has been seen in adult animals, for all preweanling ages the blood TCP exceeded the CPF concentration, and within each age group there was no evidence of non-linear kinetics over the dose range evaluated. Consistent with previous results, younger animals demonstrated a greater sensitivity to ChE inhibition as evident by the age-dependent inhibition of plasma, RBC, and brain ChE. The brain may be particularly sensitive in younger animals (i.e. PND-5) due to substantially lower levels of ChE activity relative to later preweanling stages and adults. Of particular importance was the observation that even in rats as young as PND-5, the CYP450 metabolic capacity was adequate to metabolize CPF to both TCP and CPF oxon based on the detection of TCP in blood and extensive ChE inhibition (biomarker of CPF-oxon) at all ages. In addition, the increase in the blood TCP concentration ( approximately 3-fold) in PND-17 rats relative to the response in the younger rats, are consistent with an increase in CYP450 metabolic capacity with age. This is the first reported study that evaluated both the pharmacokinetics of the parent pesticide, the major metabolite, and the extent of ChE inhibition as a function of preweanling age. The results suggest that in the preweanling rat, CPF was rapidly absorbed and metabolized, and the extent of metabolism and ChE inhibition was age-dependent. PMID- 16343728 TI - Acrylamide-induced oxidative stress and biochemical perturbations in rats. AB - Acrylamide is neurotoxic to experimental animals and humans. Also, it has mutagenic and carcinogenic effects. The present study was carried out to investigate the effects of different doses of acrylamide on some enzyme activities and lipid peroxidation in male rats. Animals were assigned at random to one of the following treatments: group 1 served as control, while groups 2, 3, 4, 5, 6 and 7 were treated with 0.5, 5, 25, 50, 250 and 500 microg/kg body weight of acrylamide, respectively in drinking water for 10 weeks. Acrylamide significantly decreased plasma protein levels and the activity of creatine kinase, while increased plasma phosphatases. The activities of transaminases and phosphatases were significantly decreased in liver and testes, while lactate dehydrogenase did not change compared to control group. Plasma and brain acetylcholinesterase activity was significantly decreased. The concentration of thiobarbituric acid reactive substances, and the activities of glutathione S transferase and superoxide dismutase in plasma, liver, testes, brain, and kidney were increased in acrylamide-treated rats. On the other hand, results obtained showed that acrylamide significantly reduced the content of sulfhydryl groups and protein in different tissues. The present results showed that different doses of acrylamide exerted deterioration effects on enzyme activities and lipid peroxidation in a dose-dependent manner. PMID- 16343729 TI - N-methyl-D-aspartate (NMDA) and the regulation of mitogen-activated protein kinase (MAPK) signaling pathways: a revolving neurochemical axis for therapeutic intervention? AB - Excitatory synaptic transmission in the central nervous system (CNS) is mediated by the release of glutamate from presynaptic terminals onto postsynaptic channels gated by N-methyl-D-aspartate (NMDA) and non-NMDA (AMPA and KA) receptors. Extracellular signals control diverse neuronal functions and are responsible for mediating activity-dependent changes in synaptic strength and neuronal survival. Influx of extracellular calcium ([Ca(2+)](e)) through the NMDA receptor (NMDAR) is required for neuronal activity to change the strength of many synapses. At the molecular level, the NMDAR interacts with signaling modules, which, like the mitogen-activated protein kinase (MAPK) superfamily, transduce excitatory signals across neurons. Recent burgeoning evidence points to the fact that MAPKs play a crucial role in regulating the neurochemistry of NMDARs, their physiologic and biochemical/biophysical properties, and their potential role in pathophysiology. It is the purpose of this review to discuss: (i) the MAPKs and their role in a plethora of cellular functions; (ii) the role of MAPKs in regulating the biochemistry and physiology of NMDA receptors; (iii) the kinetics of MAPK-NMDA interactions and their biologic and neurochemical properties; (iv) how cellular signaling pathways, related cofactors and intracellular conditions affect NMDA MAPK interactions and (v) the role of NMDA-MAPK pathways in pathophysiology and the evolution of disease conditions. Given the versatility of the NMDA-MAPK interactions, the NMDA-MAPK axis will likely form a neurochemical target for therapeutic interventions. PMID- 16343730 TI - From brainstem to cortex: computational models of saccade generation circuitry. AB - The brain circuitry of saccadic eye movements, from brainstem to cortex, has been extensively studied during the last 30 years. The wealth of data gathered allowed the conception of numerous computational models. These models proposed descriptions of the putative mechanisms generating this data, and, in turn, made predictions and helped to plan new experiments. In this article, we review the computational models of the five main brain regions involved in saccade generation: reticular formation saccadic burst generators, superior colliculus, cerebellum, basal ganglia and premotor cortical areas. We present the various topics these models are concerned with: location of the feedback loop, multimodal saccades, long-term adaptation, on the fly trajectory correction, strategy and metrics selection, short-term spatial memory, transformations between retinocentric and craniocentric reference frames, sequence learning, to name the principle ones. Our objective is to provide a global view of the whole system. Indeed, narrowing too much the modelled areas while trying to explain too much data is a recurrent problem that should be avoided. Moreover, beyond the multiple research topics remaining to be solved locally, questions regarding the operation of the whole structure can now be addressed by building on the existing models. PMID- 16343731 TI - The strong situation: a potential impediment to studying the psychobiology and pharmacology of anxiety disorders. AB - The strong situation, as formulated by social psychologists, refers to an experimental condition offering unambiguous stimuli predicting or constituting hedonically strong events that uniformly guide response sets across individuals. In relation to fear and anxiety, the strong situation results from the unambiguous threat of an imminent and dangerous stimulus that evokes the adaptive fear response among anxiety patients and healthy controls alike. The current paper describes evidence that weakening the experimental situation through reducing the certainty, temporal proximity, and/or potency of the aversive stimulus may facilitate the emergence of patient-control differences in psychobiological measures of anxious arousal. Additionally, weak situations may be useful for testing the clinical utility of anxiolytic agents, given that pharmacological treatments of anxiety disorders are not intended to reduce the adaptive, normative response likely evoked by strong threat situations. PMID- 16343732 TI - Left-right asymmetry in pelvic lymph nodes distribution: is there a right-side prevalence? AB - OBJECTIVE: To assess whether pelvic lymph nodes have a left-right asymmetric distribution. STUDY DESIGN: The oncologic databases of two gynecologic academic departments were used to identify consecutive patients undergoing pelvic systematic lymphadenectomy as part of the treatment for a variety of gynecologic malignancies. All procedures were carried out in a standardized fashion. Lymph node counts were retrieved from pathological reports. RESULTS: Four hundred and twenty-eight women underwent pelvic lymphadenectomy during the study period. The median lymph node count was higher on the right side than on the left side [10 (0 33) versus 8 (0-29); P<0.0001]. A prevalence of right-sided nodes was found in 265 (61.9%) patients, while in 44 (10.3%) cases pelvic nodes were equally distributed on the two sides. The right-sided prevalence was significantly higher than the expected 50% in each type of malignancy and surgical technique subgroup. The right-sided prevalence was statistically significant even when the analysis was performed for different nodal groups [external iliac nodes: 5 (0-23) versus 4 (0-13), P=0.005; hypogastric and obturator nodes: 6 (0-17) versus 5 (0-19), P=0.04]. Moreover, nodal count was higher on the right than on the left in obese [10 (1-33) versus 8 (1-26), P=0.0002] and nonobese women [10 (0-32) versus 9 (0 29), P<0.0001]. CONCLUSION: Our findings suggest the existence of a left-right asymmetry in pelvic lymph nodes distribution, with right-sided prevalence. PMID- 16343733 TI - Use of epidural analgesia and its relation to caesarean and instrumental deliveries-a population--based study of 94,217 primiparae. AB - OBJECTIVES: To investigate the association between epidural analgesia for labour pain relief and mode of delivery. STUDY DESIGN: The Swedish medical birth register covers 99% of all births and contains prospectively collected information from all delivery units in Sweden. The present population-based cohort study includes singleton births among nulliparae during 1998-2000, excluding deliveries with elective caesarean section, giving study population of n=94,217. The frequencies of epidural block in this population were estimated for each delivery unit. The outcomes studied were non-elective caesarean section and instrumental delivery. RESULTS: There was no clear association between frequency of epidural block and caesarean section and instrumental delivery, respectively. Delivery units with the lowest (20-29%) and the highest (60-64%) relative frequencies of epidural block had the lowest proportion of caesarean section (9.1%). For the other groups the proportion varied between 10.3 and 10.6%. Instrumental deliveries were most common, 18.8%, in delivery units with 50-59% frequency of epidural block use. The lowest incidence (14.1%) was in units using epidurals in 30-39% of cases. In the other groups (20-29, 40-49 and 60-64%) the proportion varied between 15.3 and 15.7%. CONCLUSIONS: This investigation shows no clear association between epidural use and caesarean section or instrumental delivery, indicating that there is no reason to restrict the epidural rate to improve obstetric outcome. PMID- 16343734 TI - Proximal type epithelioid sarcoma of the scrotum: a source of diagnostic confusion that needs immediate attention. AB - Epithelioid sarcoma is a rare soft-tissue sarcoma which classically presents as a subcutaneous or deep dermal mass in distal extremities of adolescents and young adults. A more aggressive "proximal" or "large-cell" type has been described in rare cases to occur as a deep soft-tissue mass at proximal body sites which tends to recur and metastasize earlier than the conventional epithelioid sarcoma. The correct diagnosis of this type is essential since these lesions can easily be misdiagnosed as other epithelioid lesions. Its prognostic factors also have not yet been fully investigated. We herein report a case of metastatic proximal-type epithelioid sarcoma arising from the scrotum of a 50-year-old man managed by radical surgery and adjuvant chemotherapy. PMID- 16343735 TI - Are organisms committed to lower their rates of entropy production? Possible relevance to evolution of the Prigogine theorem and the ergodic hypothesis. AB - The physiology at limiting and stress conditions challenges the current view that the overall reaction of metabolic processes is always far from equilibrium and, therefore, that organisms are not committed to lower their rates of entropy production. Plausibly, critical steps of natural selection takes place at limiting conditions, near equilibrium, in the linear range response of entropy production, and consequently the trend to lower the rate of entropy production could be the fitness arrow of biological evolution. The evolutionary relevance of the Prigogine theorem is discussed in connection with the ergodic hypothesis of Boltzmann. The emergence of metabolic strategies to economise carbon/energy resources, of resource-waste systems like active transport and the irreversible increase in the complexity of organisms during evolution may be consequences of a more general trend of metabolic systems to lower the rates of entropy production. PMID- 16343736 TI - DNA polymerase programmed with a hairpin DNA incorporates a multiple-instruction architecture into molecular computing. AB - Parallelism is one of the major advantages of molecular computation. A large number of data encoded in DNA molecules can be processed simultaneously by molecular biology techniques, although only a single set of instructions has been implemented in a solution. We have developed a computing machine, called the "whiplash" machine, which is made of DNA polymerase and a hairpin DNA. This machine simulates a finite state machine, executing its own instructions encoded in the DNA moiety, and would thus be applicable to multiple-instruction operation in a solution. In the present study, we explored the feasibility of this novel type of parallelism by applying the whiplash machine in a computation of the directed Hamiltonian path problem. The possible paths in a given graph were represented with different instruction sets, which were then implemented separately by whiplash machines in a test tube. After an autonomous operation of the machines, only the machine that implemented the instruction set corresponding to the Hamiltonian path was recovered from the tube. On the basis of the efficiency of machine operation, which was experimentally determined, 10(10) different instruction sets could be implemented simultaneously in a 1-ml solution. PMID- 16343737 TI - The regulated expression of c-IAP1 and c-IAP2 during the rat seminiferous epithelial cycle plays a role in the protection of germ cells from Fas-mediated apoptosis. AB - The inhibitor of apoptosis proteins, c-IAP1 and c-IAP2, are highly expressed in rat testis and potentially play a regulatory role in testicular apoptosis. To better understand their functions during spermatogenesis, we have analyzed their spatio-temporal distribution in rat testis, how their expression is controlled by the paracrine stem-cell factor (SCF) and how they affect Fas-mediated apoptosis. Both c-IAP1 and c-IAP2 showed cycles of transcriptional expression, throughout the seminiferous epithelial cycle. c-IAP1 protein showed a diffuse nuclear distribution in type B spermatogonia, preleptotene, leptotene, and zygotene spermatocytes. In pachytene spermatocytes, c-IAP1 colocalized with SUMO-1 in the XY-body. c-IAP2 protein was cytoplasmic in spermatocytes, from stage VI pachytene onwards, round spermatids, elongated spermatids and Leydig cells. Its expression was upregulated by SCF. Inhibition of IAP activity resulted in a greater sensitivity of germ cells to Fas-mediated apoptosis. These results suggest an important role for IAPs in the regulation of spermatogenic apoptosis. PMID- 16343738 TI - Multiple endocrine neoplasia type 2 and the RET protooncogene: from bedside to bench to bedside. AB - Although the initial characterization of the various MEN-2 associated phenotypes (familial medullary thyroid cancer, multiple endocrine neoplasia 2A and 2B) evolved at the bedside, it was at the bench where the underlying RET (REarranged during Transfection) germline mutations were identified. Molecular information has revolutionized our understanding and continues to transform the clinical management of this fascinating endocrine tumor syndrome of neural crest derivation, which consists of medullary thyroid cancer, pheochromocytoma, and parathyroid hyperplasia/adenoma. DNA-based identification of RET carriers did not require comprehension of the gene, but was a prerequisite for clarifying gene function and devising biologic compounds blocking RET phosphorylation. With the continuing expansion of our knowledge about the underlying molecular mechanisms and our growing therapeutic abilities, multiple endocrine neoplasia type 2 is gradually returning home to the bedside, closing the loop from bedside to bench to bedside. PMID- 16343739 TI - 11Beta-hydroxysteroid dehydrogenase type 1: purification from human liver and characterization as carbonyl reductase of xenobiotics. AB - 11Beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) catalyzes the interconversion of 11-oxo glucocorticoids to their 11-hydroxy metabolites, thereby controlling access of glucocorticoid hormones to the glucocorticoid receptor. Interestingly, evidence is emerging that 11beta-HSD1 fulfills an additional role in the metabolism of xenobiotic carbonyl compounds. In our studies, 11beta-HSD1 was identified as a microsomal reductase that initiates the final detoxification of xenobiotics by reducing them to alcohols that are easier to conjugate and eliminate. With its pluripotent substrate specificities for glucocorticoids and xenobiotics, 11beta-HSD1 adds to an expanding list of those hydroxysteroid dehydrogenases which, on the one hand, are capable of catalyzing the carbonyl reduction of non-steroidal carbonyl compounds, and which, on the other hand, exhibit great specificity to their physiological steroid substrates. It is conceivable that large interferences must occur between endogenous steroid metabolism and the detoxification of xenobiotic compounds on the level of hydroxysteroid dehydrogenases. PMID- 16343740 TI - The interleukin-10 levels as a potential indicator of positive response to interferon beta treatment of multiple sclerosis patients. AB - OBJECTIVES: Only a part of MS patients treated with interferon beta (IFN) respond positively to the applied treatment and to date no parameter predicting the response to treatment has been found. The aim of the study was to determine whether the levels of interleukin-10 and -12 (IL-10 and IL-12) might be the parameters enabling us to distinguish those patients who would best respond to therapy before the IFN treatment. PATIENTS AND METHODS: The study included 29 patients with clinically definite relapsing-remitting MS treated with IFN beta. In all of them the levels of IL-10 and IL-12 in blood serum and cerebrospinal fluid were determined before treatment using ELISA method. After the 2-year therapy the patients responding and nonresponding to IFN therapy were distinguished on the basis of clinical parameters. RESULTS: In the patients responding positively to IFN treatment the level of IL-10 in blood serum before treatment was found to be significantly lower (p<0.05) and distinctively differentiated responders from nonresponders. The IL-12 levels were similar both in cerebrospinal fluid and serum and no significant differences between responders and nonresponders were found. CONCLUSION: Our observations suggest that IL-10 level may be a useful parameter to identify the patients potentially responding to IFN therapy. PMID- 16343741 TI - Impact of genetic background on spontaneous or 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP)-induced intestinal tumorigenesis in Min/+ mice. AB - We have studied the impact of genetic background on susceptibility to spontaneous or 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced intestinal tumorigenesis. The increase in small intestinal tumor number after PhIP exposure was 3.8- and 3.7-fold above the spontaneous levels in multiple intestinal neoplasia (Min)/+ F1 mice with AKR/J and A/J backgrounds, respectively, compared with only 3-fold in C57BL/6J mice. In the colon, PhIP increased the number of tumors slightly more in C57BL/6J mice (3.3-fold) than in A/J mice (3.0-fold). AKR/J mice had no colonic tumors. Most of the tumors were located in the distal two-thirds of the small intestine in all three strains. PMID- 16343742 TI - Influence of polymorphism in DNA repair and defence genes on p53 mutations in bladder tumours. AB - We studied the effects of polymorphisms in nine genes involved in DNA repair and detoxification on occurrence and type of p53 mutation in 327 bladder cancer patients. The included polymorphisms are XPC(Lys939Gln), XPD(Lys751Gln), XPG(Asp1104His), XRCC1(Arg3999Gln), XRCC3(Thr241Met), NBS1(Glu185Gln), cyclin D1(Pro241Pro), MTHFR(Ala222Val and Glu429Ala) and NQO1(Arg139Trp and Pro187Ser). We found increased risk for p53 mutation among cyclin D1 variant allele homozygotes (OR 2.4 CI 0.8-6.7). Among non-smokers, 75% (3/4) with p53 mutation but only 12.5% (3/24) without p53 mutations were XRCC3 241Met homozygotes (P=0.03). Among smokers, all p53 transversions (3/3), but only 41.7% (5/12) of p53 transitions were found among carriers of the XPC 939Gln allele. Individuals carrying the NQO1 187Ser allele showed increased risk for p53 transversions (OR 4.7, CI 0.9-26.1). All (2/2) NQO1 139Trp allele carriers but only 17.5% (7/40) of the Arg139 homozygotes had p53 transversions. Our findings suggest that altered repair and detoxification due to genetic polymorphism may influence the occurrence of p53 mutations in bladder cancer. PMID- 16343743 TI - Analysis of genetic events in 17p13 and 9p21 regions supports predominant monoclonal origin of multifocal and recurrent bladder cancer. AB - Clonality was tested in 86 tumours from 25 patients with recurrent and multifocal superficial bladder transitional cell carcinomas (TCCs) using the analysis of TP53 mutations and of LOH in the 17p13 and 9p21 regions. Tumours from the majority of individuals showed either absence or presence of the same TP53 mutation and/or an identical LOH pattern, with the same allele lost in all tumours. Only two pairs of tumours from two patients had discordant findings, which were incompatible with monoclonality. Therefore, our results rather support the monoclonal model of development of highly recurrent superficial bladder TCCs. PMID- 16343744 TI - Role of pharmacogenetics in irinotecan therapy. AB - In the treatment of advanced colorectal cancer, irinotecan has become one of the most important drugs, despite its sometimes unpredictable adverse effects. To understand why some patients experience severe adverse effects (diarrhea and neutropenia), while others do not, the metabolic pathways of this drug have to be unraveled in detail. Individual variation in expression of several phase I and phase II metabolizing enzymes and ABC-transporters involved in irinotecan metabolism and excretion, at least partly explains the observed pharmacokinetic interpatient variability. Although the difference in expression-level of these proteins to a certain amount is explained by physiologic and environmental factors, the presence of specific genetic determinants also does influence their expression and function. In this review, the role of genetic polymorphisms in the main enzyme-systems (carboxylesterase, cytochrome P450 3A, and uridine diphosphate-glucuronosyltransferase) and ABC-transporters (ABCB1, ABCC2, and ABCG2) involved in irinotecan metabolism, are discussed. Since at this moment the field of pharmacogenetics and pharmacogenomics is rapidly expanding and simultaneously more rapid and cost-effective screening methods are emerging, a wealth of future data is expected to enrich our knowledge of the genetic basis of irinotecan metabolism. Eventually, this may help to truly individualize the dosing of this (and other) anti-cancer agent(s), using a personal genetic profile of the most relevant enzymes for every patient. PMID- 16343745 TI - Evaluation of ettringite and hydrocalumite formation for heavy metal immobilization: literature review and experimental study. AB - The immobilization of heavy metal oxyanions like chromate, arsenate and selenate, has proven to be a challenging task as they are highly mobile in alkaline environments involving S/S of contaminated media. Ettringite, a pozzolanic phase that forms in cementitious materials, has been proposed as a viable immobilization mechanism for oxyanions, wherein the oxyanion may substitute for sulfate in the ettringite structure. A literature review on the immobilization potential of ettringite showed that the substitution potential exists from the thermodynamic point of view where the formation of substituted ettringites occurs under strictly controlled conditions. The pH control over a narrow range is essential for ettringite stability; it becomes even narrower for substituted ettringites, as competing effects with sulfate ettringite and monophases are significantly affected by pH. The presence of sulfate has a catalytic effect on oxyanion incorporation in ettringite. Rapid leaching may occur when the treated media is exposed to sulfate influx. Conversely, monophases seem to be more suitable than ettringite for oxyanion immobilization, mainly as they control oxyanion solubility to lower levels than ettringite. A shift to the thermodynamic equilibrium caused by a shift in environmental conditions may result in monophase conversion to ettringite, which may lead to catastrophic expansion, as widely demonstrated in the cement and soils literature. Overall, the sensitivity of phase equilibria in cement-like systems involving oxyanions is significant with regard to multiple parameters and it is uncertain to what extent these can be predicted and/or controlled in the field. PMID- 16343746 TI - A review of tungsten: from environmental obscurity to scrutiny. AB - Since its discovery, tungsten, a transition element of Group VIb of the Periodic Table of Elements, and its compounds have been considered environmentally benign. Its presence in biological and drinking water samples in Fallon, Nevada, an acute lymphocytic leukemia cluster struck community has alarmed public health, environmental and regulatory agencies. Tungsten, a metal of extraordinary properties that make it hardly substitutable, is considered an essential commodity with a wide variety of uses stretching from household necessities to highly specialized applications. This work is undertaken in order to explore aspects of environmental behavior of tungsten and its compounds. Occurrence data in terrestrial, atmospheric, aquatic and biotic systems are presented. Various aspects of environmental chemistry, fate transport across environmental interfaces and toxicology are discussed with the objective of identifying knowledge gaps and outlining directions for future research. PMID- 16343747 TI - A study on the chemical and mineralogical characterization of MSWI fly ash using a sequential extraction procedure. AB - The presence of heavy metals in municipal solid waste incineration (MSWI) fly ash is of environmental concern due to their leaching potential in landfill environments. Sequential chemical extraction was performed on fly ash samples from a large-scale municipal solid waste incineration plant in East China. The transformation of the mineralogical species of fly ash during the sequential extraction was studied using X-ray fluorescence (XRF) and X-ray powder diffraction (XRD). The leaching behavior of heavy metals such as zinc, lead, cadmium and copper in MSWI fly ash was considered to have a dependency relationship with the components of calcium, such as aphthitalite, calcite, anhydrite and calcium aluminate or calcium aluminosilicate. PMID- 16343748 TI - Adsorption of bisphenol-A from aqueous solution onto minerals and carbon adsorbents. AB - The adsorption behaviors of bisphenol-A, which has been listed as one of endocrine disrupting chemicals, from aqueous solution onto four minerals including andesite, diatomaceous earth, titanium dioxide, and activated bleaching earth, and two activated carbons with coconut-based and coal-based virgins were examined in this work. Based on the adsorption results at the specified conditions, the adsorption capacities of activated carbons are significantly larger than those of mineral adsorbents, implying that the former is effective for removal of the highly hydrophobic adsorbate from the aqueous solution because of its high surface area and low surface polarity. The adsorption capacities of bisphenol-A onto these mineral adsorbents with different pore properties are almost similar in magnitude mainly due to the weakly electrostatic interaction between the mineral surface with negative charge and the target adsorbate with hydrophobic nature. Further, a simplified kinetic model, pseudo-second-order, was tested to investigate the adsorption behaviors of bisphenol-A onto the two common activated carbons at different solution conditions. It was found that the adsorption process could be well described with the pseudo-second-order model. The kinetic parameters of the model obtained in the present work are in line with the pore properties of the two adsorbents. PMID- 16343749 TI - Degradation of Procion Red H-E7B reactive dye by coupling a photo-Fenton system with a sequencing batch reactor. AB - A bench-scale study combining photo-Fenton reaction with an aerobic sequencing batch reactor (SBR) to degrade a commercial homo-bireactive dye (Procion Red H E7B, 250mgl(-1)) was investigated. The photo-Fenton process was applied as a pre treatment, avoiding complete mineralisation, just to obtain a bio-compatible water able to be treated by means of the SBR in a second step. In this sense, different Fenton reagent concentrations were assessed by following dye solution biodegradability enhancement (BOD(5)/COD), as well as the toxicity (EC(50)), DOC, colour (Abs(543.5)) and H(2)O(2) evolution with photo-Fenton irradiation time. Obtained pre-treated solutions were biologically oxidized in a SBR containing non acclimated activated sludge. Different hydraulic retention time (HRT) in the bioreactor were tested to attain the maximum organic load removal efficiency. Best results were obtained with 60min of 10mgl(-1) Fe(II) and 125mgl(-1) H(2)O(2) photo-Fenton pre-treatment and 1 day HRT in SBR. PMID- 16343750 TI - Thermal hazard of iron picrate. AB - Iron picrate (FePic) was synthesized under conditions similar to those that result in the natural deterioration of chemical weapons. Its thermal hazard was investigated by comparing it with iron picrate obtained by the chemical synthesis method (FePic(Ba)). FePic has eight or more water molecules of crystallization and consists of a mixture of various hydrates. It shows low sensitivity to friction and drop hammer tests due to the large number of water molecules of crystallization. Under the experimental conditions of the thermal analysis, the hydrated iron picrates began to decompose before being dehydrated to form the anhydrous salt. Prolonged holding under natural environmental conditions does not appear to result in formation of the dangerous anhydrous salt. Based on the observed heating rate dependence, it is thought that the hydrated iron picrate should start to decompose before dehydration to the anhydrous salt occurs when subjected to a large heating rate. PMID- 16343751 TI - Possible production of ceramic tiles from marine dredging spoils alone and mixed with other waste materials. AB - Dredging spoils, due to their composition could be considered a new potential source for the production of monolithic ceramics. Nevertheless, abundance of coloured oxides in these materials preclude the possibility of obtaining white products, but not that of producing ceramics with a good mechanical behaviour. As goal of the present research we have produced and studied samples using not only dredging spoils alone, but also mixtures with other waste materials such as bottom ashes from an incinerator of municipal solid waste, incinerated seawage sludge from a municipal seawage treatment plant and steelworks slag. Blending of different components was done by attrition milling. Powders were pressed into specimens which were air sintered in a muffle furnace and their shrinkage on firing was determined. Water absorption, density, strength, hardness, fracture toughness, thermal expansion coefficient of the fired bodies were measured; XRD and SEM images were also examined. The fired samples were finally tested in acidic environment in order to evaluate their elution behaviour and consequently their environmental compatibility. It is observed that, although the shrinkage on firing is too high for the production of tiles, in all the compositions studied the sintering procedure leads to fine microstructures, good mechanical properties and to a limitation of the release of many of the most hazardous metals contained in the starting powders. PMID- 16343752 TI - The AETL methodology as a potential solution to current challenges associated with the development and use of acute exposure levels in Seveso II applications. AB - This paper analyses current trends in the development and use of acute exposure levels in Europe for the implementation of the Seveso II Directive [Council Directive 96/82/EC of December 9, 1996 on the control of major-accident hazards involving dangerous substances. Official Journal of the European Communities, vol. L 10, January 14, 1997, Luxembourg, pp. 13-33]. It also describes a new initiative to develop a European methodology for deriving acute exposure threshold levels that responds to emerging needs in this area. The need for acute exposure values to predict human health effects of potential accidents on exposed populations has burgeoned in recent years. As the driving legislation for managing industrial hazards in Europe, the Seveso II Directive has particularly influenced this trend. Yet at this time it is questionable whether the availability and range of acute exposure values for toxic substances has kept apace with the growing need. Results of a survey of Seveso II competent authorities in the EU-15 revealed that a variety of different types of acute exposure values (AEGLs, EPRGs, etc.) are used for Seveso II applications. Moreover, a comparison of these values indicates gaps in coverage of substances as well as inconsistencies in terms of how health effects and exposure periods are defined for each type. These findings highlight an opportunity for greater collaboration on scientific inputs to application of the Directive in Europe. The ACUTEX project is an EU-funded research project aimed at furthering scientific exchange and collaboration in support of the development of acute exposure levels for toxic substances in Europe. Its goal is to develop a European methodology for deriving acute exposure threshold levels (AETLs). In particular, it provides the possibility for a common European platform for developing additional acute exposure values to meet emerging needs and cover more chemical substances. To maximise success, the work plan is designed to meet two very important challenges, the need to complement and add value to the existing array of acute exposure methodologies and the necessity of meeting requirements of a diverse range of European stakeholders. As such the project will draw on collaboration among European scientists and process of deliberation among stakeholders to deliver the following key results: (1) to facilitate wide acceptance of the methodology in Europe by both the scientific community and communities of different end-users; (2) to provide greater equivalence and transparency in implementation of the Seveso II Directive across the Member States, specifically through the development of common scientific bases for assessing risks and making risk management decisions related to toxic releases; (3) to produce a methodology that remains open to future collaboration on derivation of acute exposure levels on a European and a global basis. PMID- 16343753 TI - Factors influencing the removal of fluoride from aqueous solution by calcined Mg Al-CO3 layered double hydroxides. AB - Layered double hydroxides (LDH) calcined at different temperatures (denoted as CLDH) have been demonstrated to recover their original layered structure in the presence of appropriate anions. In the light of this so-called "memory effect", a study of removal of fluoride from aqueous solution by calcined Mg-Al-CO(3)-LDH has been carried out. The LDH calcined at 500 degrees C had the highest capacity of removal of fluoride ion, because of retention of its intrinsic structure. The CLDH with an Mg/Al ratio of 2 has a remarkable ability to adsorb anions. The adsorption loading is higher for the calcined Mg-Al-LDH than for calcined Zn-Al and Ni-Al-LDH. The influence of varying the conditions for removal of fluoride, such as the pH of aqueous solution, the initial fluoride concentration, the dosage of adsorbent, and temperature on removal of fluoride have been investigated. The influence of co-existing anions in fluoride aqueous solution indicates that the percentage of removal of fluoride increased in order PO(4)(3-) < Cl(-) approximately SO(4)(2-) < Br(-) << NO(3)(-). It was found that maximum removal of fluoride from aqueous solutions was obtained in 6h at pH 6.0 with an initial concentration of 50 mg/L, and that the retention of fluoride ions by the CLDH material was 98% or higher. The residual fluoride concentration was found to be 0.4 mg/L with an initial concentration of 20 mg/L, which meets the national standard for drinking water quality. The Freundlich isotherm and Langmuir isotherm were used to fit the data of equilibrium experiments. The results of X ray diffraction, FT-IR and TG-MS demonstrate that the adsorption phenomenon is accompanied by rehydration with concomitant uptake of fluoride ions to rebuild the initial layered structure. PMID- 16343754 TI - The reuse of dried activated sludge for adsorption of reactive dye. AB - Adsorption processes are alternative effective methods for removal of textile dyes from aqueous solutions. The adsorption ability of adsorbent affects by physico-chemical environment for this reason in this paper effect of initial pH, dye concentrations, temperature and dye hydrolyzation were determined in a batch system for removal of reactive dye by dried activated sludge. The Langmuir isotherm model was well described of adsorption reactive dye and maximum monolayer adsorption capacity (at pH 2) of activated sludge was determined as 116, 93 and 71mgg(-1) for 20 degrees , 35 degrees and 50 degrees C, respectively. Initial pH 2, 20 degrees C and 30min contact time are suitable for removal of reactive dyes from aqueous solutions. Activated sludge was characterized by FT-IR analysis and results showed that active sludge has different functional groups and functional groups of activated sludge are able to react with dye molecules in aqueous solution. The pseudo first-order, second-order and intraparticle diffusion kinetics were used to describe the kinetic data. The pseudo second order kinetic model was fit well over the range of contact times and also an intra particle diffusion kinetic model was fit well but in the first 30min. The dye hydrolyzation was affected adsorption capacity of biomass and adsorption capacity of biomass decreased with dye hydrolyzation from 74 to 38mgg(-1). PMID- 16343755 TI - Kinetic and safety assessment for salicylic acid nitration by nitric acid/acetic acid system. AB - The nitration process of salicylic acid for the production of the important intermediate 5-nitrosalicylic acid is studied from thermokinetic and safety points of view. Investigations carried out by considering, as process deviations, the loss of the thermal control point out the possibility of runaway phenomena due to the occurrence of polynitration reactions. Isothermal experiments are carried out in various conditions to assess the involved reaction network and reaction kinetics. PMID- 16343756 TI - Effect of chaotic mixing on enhanced biological growth and implications for wastewater treatment: a test case with Saccharomyces cerevisiae. AB - Mixing patterns and modes have a great influence on the efficiency of biological treatment systems. A series of laboratory experiments was conducted with a controlled, small-scale analog of a pilot wastewater aeration tank, consisting of two eccentrically placed cylinders. By controlling the rotation direction and speed of the two cylinders, it has been possible to develop chaotic flow fields in the space between the walls of the cylinders. Our experiments utilized Saccharomyces cerevisiae as the biological oxidation organism and air bubbles as the mixing agent supplied by a large fine pore diffuser to the cells in their exponential growth phase. The effect of various mixing patterns on cell growth was studied at different cylinder eccentricities, rotation directions and speeds. It was found that chaotic advection flow patterns: (a) enhanced growth, and (b) sped up the onset of maximal growth of the organism by 15-18% and 14-20%, respectively. PMID- 16343757 TI - Combined effect of Bond and capillary numbers on hydrocarbon mobility in water saturated porous media. AB - The mobilization of an oil bank under the combined effect of Bond (N(B)) and capillary (N(C)) numbers, in a packed bed column of glass beads saturated with water, has been investigated. In order to reach the irreducible saturation the experiments have been run with sweeping water velocities outside the range of validity of the Darcy's law. The size of the glass beads was varied in the range between 2 mm and 5 mm. The oils used for the tests are hexadecane and hexane with viscosities different for an order of magnitude and densities smaller than that of water, and alpha-methylnaphthalene, which has a density very close to that of water, in order to single out the effect of the capillary number on the mobilization process. The plots of oil saturation as function of the trapping number (N(T)), which is the vectorial sum of N(B) and N(C), are reported and a mobilization diagram is drawn. Furthermore, a few tests in a basin, simulating an aquifer at a laboratory scale, have proved that the results obtained in the packed column are useful for determining the fate of a spill of oil above an aquifer. For these experiments also perchloroethylene (PCE), which has a density greater than that of water, has been used. PMID- 16343758 TI - Adsorption of dimethyl methylphosphonate on metal impregnated carbons under static conditions. AB - Active carbon, grade 80 CTC, of surface area 1199m(2)/g, 12x30 BSS particle size and coconut shell origin was impregnated (5%, W/W) with various impregnants such as Cu(II) 1,1,1,5,5,5-hexafluoroacetylacetonate, Cu(II) 1,1,1 trifluoroacetylacetonate, 1-phenylbute-1,3-dione-2-oxime plus Cu(II) using incipient wetness technique. These impregnated carbons along with active carbon (Grade 80 CTC) and whetlerite were studied for the adsorption of dimethyl methylphosphonate (DMMP) at 33+/-1 degrees C under static conditions. Cu(II) 1,1,1,5,5,5-hexafluoroacetylacetonate impregnated carbon system showed highest uptake (68.5%, W/W) of DMMP amongst all the carbon systems, however, active carbon with higher surface area could adsorb 61.5% (W/W) of DMMP under same conditions. It indicated that the adsorption by Cu(II) 1,1,1,5,5,5 hexafluoroacetylacetonate impregnated carbon was not only due to physisorption but chemisorption as well. Kinetics of adsorption was also studied and various parameters such as equilibration time, equilibration capacity, rate constant (k), diffusional exponent (n) and constant (K) were determined. Carbons with and without DMMP exposure were also studied using IR and TGA techniques. Reaction products were analyzed using gas chromatography coupled with mass spectrometry (GC/MS) and found to be methyl methylphosphonic acid (MMPA) and methylphosphonic acid (MPA) for Cu(II) 1,1,1,5,5,5-hexafluoroacetylacetonate impregnated carbon. PMID- 16343759 TI - Adsorption of cationic polyacrylamide onto sepiolite. AB - The adsorption of PAM onto sepiolite from aqueous solutions has been investigated systematically as a function of some parameters such as calcination temperature of sepiolite, pH, ionic strength and temperature. The adsorption of cationic polyacrylamide (PAM) increases with pH from 5.50 to 11.00, temperature from 25 to 55 degrees C and ionic strength from 0 to 0.1molL(-1). The sepiolite sample calcined at 200 degrees C has a higher adsorption capacity than the other calcined samples. Adsorption isotherms of PAM onto sepiolite have been determined and correlated with common isotherm equations such as Langmuir and Freundlich isotherm models. The Langmuir isotherm model appeared to fit the isotherm data better than the Freundlich isotherm model. The physical properties of this adsorbent are consistent with the parameters obtained from the isotherm equations. The zeta potentials of sepiolite suspensions have been measured in aqueous solutions of NaCl and different PAM concentrations and pH. From the experimental results: (i) pH strongly alters the zeta potential of sepiolite, (ii) sepiolite has an isoelectric point at about pH 6.6 in water and about pH 8 in 250mgL(-1) PAM concentration, (iii) PAM changes the interface charge from negative to positive for sepiolite. Effect of temperature on adsorption has been quantified by calculating various thermodynamic parameters such as Gibbs free energy, enthalpy and entropy. The dimensionless separation factor (R(L)) has shown that sepiolite can be used for adsorption of PAM from aqueous solutions. PMID- 16343760 TI - Adsorption of herbicide paraquat by clay mineral regenerated from spent bleaching earth. AB - The adsorption of herbicide paraquat (as model adsorbate) in aqueous solution onto regenerated clay mineral from bleaching earth waste has been studied in a batch reaction system. The adsorption rate has been investigated under the controlled process parameters including initial pH, salinity and temperature. Based on the high affinity between cationic paraquat and clay mineral, a pseudo second order model has been developed using experimental data to predict the rate constant of adsorption, and equilibrium adsorption capacity. The results showed that the adsorption process could be satisfactorily described with the reaction model and were reasonably explained by assuming a competitive adsorption mechanism in the ion exchange process. Further, the fitted adsorption capacity at equilibrium decreased with increasing temperature. It implied that the strong interaction might play an important role in the paraquat-clay system. Overall, the results from this study demonstrated that the clay resource regenerated from bleaching earth waste could be used as a low-cost mineral adsorbent for the removal of environmental cationic organic pollutants from the aqueous solution. PMID- 16343761 TI - Abiotic oxidation of catechol by soil metal oxides. AB - The mechanism of catechol oxidation by soil metal oxides is investigated in a slurry reactor. This abiotic transformation is shown to consist in a three-step process. The first step is a heterogeneous reaction. Catechol undergoes fast, partial oxidation at the expenses of Fe and Mn oxides contained in the soil. In the second step, reduced Fe and Mn are released into the aqueous solution and immediately complexed by catechol. Metal-catecholate complexes are stable at the very low dissolved-oxygen concentration levels attained under nitrogen sparging. The third step is a homogenous reaction. The highly reactive intermediate produced by catechol partial oxidation initiates catechol polymerisation. Under nitrogen sparging, the polymerisation process ends rather rapidly, thus yielding only partial conversion of the phenol and producing low-molecular weight, water soluble polymers. Further oxidation of the metal-catecholate complexes formed in the second step only occurs under air sparging. Thus, reactive intermediates are formed at much higher concentration levels than those attained when nearly no oxygen is present in solution. The polymerisation proceeds at a much faster rate until, under the experimental conditions adopted, complete catechol conversion is attained and high-molecular-weight, insoluble polymers are produced. PMID- 16343762 TI - Improving alachlor biodegradability by ferrate oxidation. AB - Alachlor can be recalcitrant when present at high concentrations in wastewater. Ferrate oxidation was used as a pretreatment to improve its biodegradability and was evaluated by monitoring alachlor elimination and removal of COD(Cr) (chemical oxygen demand determined by potassium dichromate) during the oxidation process up to a value compatible with biological treatment. Ferrate oxidation resulted in elimination of alachlor followed by degradation of its intermediates. High pH suppressed alachlor removal and COD(Cr) removal due to the low redox potential of ferrate ions. Although alachlor can be totally eliminated within 10 min under optimized conditions (alachlor, 40 mg l(-1); ferrate:alachlor molar ratio, 2; and pH 7.0), its complete mineralization cannot be achieved by ferrate oxidation alone. Alachlor solution treated by ferrate for 10 min inhibited an up-flow biotreatment with activated sludge. The biodegradability of ferrate-pretreated solution improved when the treatment was increased to 20 min, at the point of which BOD(5)/COD(Cr) ratio of the treated solution was increased to 0.87 from 0.35 after 10 min treatment. Under optimized conditions, ferrate oxidation for 20 min resulted in total elimination of alachlor, partial removal of COD(Cr) and the ferrate-treated solution could be effectively treated by the up-flow activated sludge process. PMID- 16343763 TI - The effect of some operating variables on the adsorption of lead and cadmium ions on kaolinite clay. AB - Modification of kaolinite clay mineral with orthophosphate (p-modified sample) enhanced adsorption of Pb and Cd ions from aqueous solutions of the metal ions. Increasing pH of solutions of metal ions, increasing adsorbent dose and increasing concentration of metal ion, increased the adsorption of metal ions. Adsorption of both metal ions simultaneously on both unmodified and p-modified samples indicates that adsorption of one metal ion is suppressed to some degree by the other. The presence of electrolyte and their increasing concentration reduced the adsorption capacities of both unmodified and p-modified samples for the metal ions. Ca-electrolytes had more negative effect on the adsorption capacities of the adsorbents than Na-electrolytes. Ca-electrolytes reduced adsorption capacities of the adsorbents for Pb and Cd ions. From Langmuir plots it was observed that these electrolytes increased the binding energy constant of the metal ions unto the adsorbents especially on the p-modified samples. The rate of adsorption of Pb and Cd ions on p-modified adsorbent were increased and equilibrium of metal ion solution were more quickly reached (8min for Pb ions and 12min for Cd ions) with p-modified adsorbent as against 20min for adsorption of both metal ions on unmodified adsorbent when 200mg/L of metal ion solutions were used during the kinetic studies. When adsorption data were fitted against Langmuir, Freundlich, Toth and Langmuir-Freundlich isotherms, satisfactory fits were found with the Freundlich isotherm. However, at low concentration of metal ions, data also showed satisfactory fits to Langmuir isotherm. PMID- 16343764 TI - Bioremediation of Pb-contaminated soil by incubating with Phanerochaete chrysosporium and straw. AB - The bioremediation of the simulated lead (Pb)-contaminated soils by incubating with Phanerochaete chrysosporium and straw was studied at laboratory-scale. The soil pH, Pb concentration, soil microbial biomass, microbial metabolic quotient, microbial quotient and microbial biomass C-to-N ratios were monitored. The above indicators were to study the stress of Pb on soil and the microbial effects during the bioremediation process. It was found that the soils treated with P. chrysosporium and straw showed a much lower concentration of soluble-exchangeable Pb, lower metabolic quotient and biomass C-to-N ratios (0mgkg(-1) dry weight soil, 1.9mg CO(2)-Cmg(-1) biomass carbon and 4.9 on day 60, respectively) and higher microbial biomass and microbial quotient (2258mgkg(-1) dry weight soil and 7.86% on day 60, respectively) compared with the controls. In addition, the kinetic parameters in the model based on logistic equation were calculated by the BIOLOG data. By analyzing those kinetic parameters some information on the metabolic capacity of the microbial community could be obtained. All the results indicated that the bioavailability of Pb in contaminated soil was reduced so that the potential stress of Pb was alleviated, and also showed that the soil microbial effects and the metabolic capacity of microbial community were improved. PMID- 16343765 TI - Consensus oriented fuzzified decision support for oil spill contingency management. AB - Studies on multi-group multi-criteria decision-making problems for oil spill contingency management are in their infancy. This paper presents a second-order fuzzy comprehensive evaluation (FCE) model to resolve decision-making problems in the area of contingency management after environmental disasters such as oil spills. To assess the performance of different oil combat strategies, second order FCE allows for the utilization of lexical information, the consideration of ecological and socio-economic criteria and the involvement of a variety of stakeholders. On the other hand, the new approach can be validated by using internal and external checks, which refer to sensitivity tests regarding its internal setups and comparisons with other methods, respectively. Through a case study, the Pallas oil spill in the German Bight in 1998, it is demonstrated that this approach can help decision makers who search for an optimal strategy in multi-thread contingency problems and has a wider application potential in the field of integrated coastal zone management. PMID- 16343766 TI - Comment on "Remediation of soil contaminated with the heavy metal (Cd2+)". PMID- 16343767 TI - Quantum chemical, ballistic and explosivity calculations on 2,4,6,8-tetranitro 1,3,5,7-tetraaza cyclooctatetraene: a new high energy molecule. AB - Ab initio molecular orbital calculations have been carried out on 2,4,6,8 tetranitro-1,3,5,7-tetraazacyclooctatetraene, the tetramer of the series (NO(2)CN)(n) where n=1-4, using the Hartree-Fock theory with the 6-31 G(d) basis set. These calculations yield three conformers for the tetramer with D(4h), C(4h) and C(2) symmetries. The nonplanar conformer with the C(2) symmetry turns out to be 99.0 and 164.4kJmol(-1), respectively, lower in energy than the C(4h) and D(4h) conformers. The electron density topography - the density at the bond critical point - has been used as a measure of the CNO(2) strengths. Based on these bond strengths, heats of formation [obtained from the parametric model 3 (PM3) method] and specific decomposition energies, it may be concluded that (NO(2)CN)(4) is a promising candidate in the class of high energy molecules. Theoretically computed explosive (velocity of detonation, detonation pressure, etc.) and ballistic (characteristic velocity, specific impulse, etc.) parameters support these conclusions. PMID- 16343768 TI - Opioid receptor-mediated hyperalgesia and antinociceptive tolerance induced by sustained opiate delivery. AB - Opiates are commonly used to treat moderate to severe pain and can be used over prolonged periods in states of chronic pain such as those associated with cancer. In addition, to analgesic actions, studies show that opiate administration can paradoxically induce hyperalgesia. At the pre-clinical level, such hyperalgesia is associated with numerous pronociceptive neuroplastic changes within the primary afferent fibers and the spinal cord. In rodents, sustained opiate administration also induces antinociceptive tolerance. The mechanisms by which prolonged opiate exposure induces hyperalgesia and the relationship of this state to antinociceptive tolerance remain unclear. The present study was aimed at determining whether sustained opiate-induced hyperalgesia, associated neuroplasticity and antinociceptive tolerance are the result of specific opiate interaction at opiate receptors. Enantiomers of oxymorphone, a mu opioid receptor agonist, were administered to rats by spinal infusion across 7 days. Sustained spinal administration of (-)-oxymorphone, but not its inactive enantiomer (+) oxymorphone or vehicle, upregulated spinal dynorphin content, produced thermal and tactile hypersensitivity, and produced antinociceptive tolerance. These results indicate that these pronociceptive actions of sustained opiate administration require specific interaction with opiate receptors and are unlikely to be the result of accumulation of potentially excitatory metabolic products. While the precise mechanisms, which may account for these pronociceptive changes remain to be unraveled, the present data point to plasticity initiated by opiate receptor interaction. PMID- 16343769 TI - Effect of neurofeedback training on the neural substrates of selective attention in children with attention-deficit/hyperactivity disorder: a functional magnetic resonance imaging study. AB - Attention Deficit Hyperactivity Disorder (AD/HD) is a neurodevelopmental disorder mainly characterized by impairments in cognitive functions. Functional neuroimaging studies carried out in individuals with AD/HD have shown abnormal functioning of the anterior cingulate cortex (ACC) during tasks involving selective attention. In other respects, there is mounting evidence that neurofeedback training (NFT) can significantly improve cognitive functioning in AD/HD children. In this context, the present functional magnetic resonance imaging (fMRI) study was conducted to measure the effect of NFT on the neural substrates of selective attention in children with AD/HD. Twenty AD/HD children- not taking any psychostimulant and without co-morbidity-participated to the study. Fifteen children were randomly assigned to the Experimental (EXP) group (NFT), whereas the other five children were assigned to the Control (CON) group (no NFT). Subjects from both groups were scanned 1 week before the beginning of the NFT (Time 1) and 1 week after the end of this training (Time 2), while they performed a Counting Stroop task. At Time 1, for both groups, the Counting Stroop task was associated with significant loci of activation in the left superior parietal lobule. No activation was noted in the ACC. At Time 2, for both groups, the Counting Stroop task was still associated with significant activation of the left superior parietal lobule. This time, however, for the EXP group only there was a significant activation of the right ACC. These results suggest that in AD/HD children, NFT has the capacity to normalize the functioning of the ACC, the key neural substrate of selective attention. PMID- 16343770 TI - Leptin induces nitric oxide synthase type II in C6 glioma cells. Role for nuclear factor-kappaB in hormone effect. AB - Astrocytes in the CNS produce inflammatory mediators in response to several stimuli and cytokines. Here we investigated the in vitro effect of leptin on inducible nitric oxide synthase (iNOS) expression in a glioma cell line (C6). After hormone stimulation, culture media were analysed for accumulated stable oxidation products of NO (NO2(-) and NO3(-), designated as NO(x)), cellular RNA was extracted to determine iNOS mRNA level by RT-PCR and cellular lysates were prepared for protein expression. Leptin induced a concentration-dependent increase of NO release, related to iNOS induction. This effect was potentiated by IFN-gamma, or TNF-alpha, or IFN-gamma plus IL-1beta. Pyrrolidine dithiocarbamate (PDTC) and N-alpha-tosyl-L-lysine chloromethyl ketone (TLCK), two inhibitors of NF-kappaB activation, as well as the specific proteasome inhibitor MG132, blocked leptin-induced iNOS. The role of NF-kappaB was also confirmed by time course studies on degradation of IkappaB-alpha, which began to degrade 5 min after treatment with leptin and returned to basal level after 30-60 min. Pre-incubation of cells with MG132 inhibited leptin-induced IkappaB-alpha degradation. These results confirm the pro-inflammatory role of leptin and identify it as a potential up-regulator of cytokine-induced inflammatory response in the CNS. PMID- 16343771 TI - Comparison of spatiotemporal cortical activation pattern during visual perception of Korean, English, Chinese words: an event-related potential study. AB - The aim of this study was to compare spatiotemporal cortical activation patterns during the visual perception of Korean, English, and Chinese words. The comparison of these three languages offers an opportunity to study the effect of written forms on cortical processing of visually presented words, because of partial similarity/difference among words of these languages, and the familiarity of native Koreans with these three languages at the word level. Single-character words and pictograms were excluded from the stimuli in order to activate neuronal circuitries that are involved only in word perception. Since a variety of cerebral processes are sequentially evoked during visual word perception, a high temporal resolution is required and thus we utilized event-related potential (ERP) obtained from high-density electroencephalograms. The differences and similarities observed from statistical analyses of ERP amplitudes, the correlation between ERP amplitudes and response times, and the patterns of current source density, appear to be in line with demands of visual and semantic analysis resulting from the characteristics of each language, and the expected task difficulties for native Korean subjects. PMID- 16343772 TI - Neuromagnetic recordings reveal the temporal dynamics of auditory spatial processing in the human cortex. AB - In an attempt to delineate the assumed 'what' and 'where' processing streams, we studied the processing of spatial sound in the human cortex by using magnetoencephalography in the passive and active recording conditions and two kinds of spatial stimuli: individually constructed, highly realistic spatial (3D) stimuli and stimuli containing interaural time difference (ITD) cues only. The auditory P1m, N1m, and P2m responses of the event-related field were found to be sensitive to the direction of sound source in the azimuthal plane. In general, the right-hemispheric responses to spatial sounds were more prominent than the left-hemispheric ones. The right-hemispheric P1m and N1m responses peaked earlier for sound sources in the contralateral than for sources in the ipsilateral hemifield and the peak amplitudes of all responses reached their maxima for contralateral sound sources. The amplitude of the right-hemispheric P2m response reflected the degree of spatiality of sound, being twice as large for the 3D than ITD stimuli. The results indicate that the right hemisphere is specialized in the processing of spatial cues in the passive recording condition. Minimum current estimate (MCE) localization revealed that temporal areas were activated both in the active and passive condition. This initial activation, taking place at around 100 ms, was followed by parietal and frontal activity at 180 and 200 ms, respectively. The latter activations, however, were specific to attentional engagement and motor responding. This suggests that parietal activation reflects active responding to a spatial sound rather than auditory spatial processing as such. PMID- 16343773 TI - RANTES, MDC and SDF-1alpha, prevent the HIVgp120-induced food and water intake decrease in rats. AB - Human immunodeficiency virus (HIV)-wasting syndrome might be facilitated by the HIVgp120 affecting the immunological system. We studied the effect (subchronic administration: 5 days) of HIVgp120, and a few immune-response mediators: regulated upon activation normal T-cell expressed and presumably secreted (RANTES), stromal derived factor-1alpha (SDF-1alpha), macrophage-derived chemokine (MDC), and their combination, on food and water intake in rats, motor control and pain perception. Eighty male adult Wistar rats received an intracerebroventricular (icv) administration of: vehicle 5 microl/day or 0.92 nmol daily of HIVgp120IIIB, RANTES, SDF-1alpha, or MDC, and the combination of RANTES+HIVgp120IIIB, SDF-1alpha+HIVgp120IIIB, or MDC+HIVgp120IIIB. Food and water intake was measured every day during administration, and 24 and 48 h after the last administration. Rats were also weighed the first and the last day of experiment in order to detect the impact of these treatments in the body weight. HIVgp120IIIB significantly decreased food and water intake. These rats gain less weight than the control (vehicle) and chemokines-treated subjects with exception of those treated with SDF-1alpha that also gain less weight. In addition, HIVgp120 deteriorated motor control. HIVgp120IIIB effects on food and water intake, and motor control were prevented by these chemokines. HIVgp120+RANTES, HIVgp120+SDF-1alpha, and SDF-1alpha alone induced hyperalgesia. Results suggest an interaction between HIVgp120 and the chemokine system to generate the HIV wasting syndrome, the motor abnormalities and changes in pain perception. PMID- 16343774 TI - Electron holography of thin amorphous carbon films: measurement of the mean inner potential and a thickness-independent phase shift. AB - The phase shift induced by thin amorphous carbon films with thicknesses between 1 and 16 nm was measured by electron holography in a transmission electron microscope. Large phase shifts Delta phi are observed as the thickness of the amorphous C films decreases which cannot be described by the well-known equation Delta phi = CE V0t (V0: mean inner Coulomb potential of the material, t: sample thickness). Data plotted in a Delta phi vs. t diagram can be well-fitted by a modified equation Delta phi=CE V0t + phi add. The mean inner Coulomb potential of the amorphous carbon with a density of 1.75 g/cm3 was determined to be 9.09 V which is consistent with previous experimental data for amorphous carbon with a higher density. The thickness-independent phase offset phi(add) of 0.497 rad is large for amorphous carbon under the given experimental conditions. We suggest that a surface-related electrostatic potential is responsible for the thickness independent contribution phi add. PMID- 16343775 TI - Glycoform-dependent conformational alteration of the Fc region of human immunoglobulin G1 as revealed by NMR spectroscopy. AB - The Fc portion of immunoglobulin G (IgG) expresses the biantennary complex type oligosaccharides at Asn297 of the C(H)2 domain of each heavy chain with microheterogeneities depending on physiological and pathological states. These N glycans are known to be essential for promotion of proper effector functions of IgG such as complement activation and Fcgamma receptor (FcgammaR)-mediated activities. To gain a better understanding of the role of Fc glycosylation, we prepared a series of truncated glycoforms of human IgG1-Fc and analyzed their interactions with human soluble FcgammaRIIIa (sFcgammaRIIIa) and with staphylococcal protein A by surface plasmon resonance and nuclear magnetic resonance (NMR) methods. Progressive but less pronounced reductions in the affinity for sFcgammaRIIIa were observed as a result of the galactosidase and subsequent N-acetylhexosaminidase treatments of IgG1-Fc. The following endoglycosidase D treatment, giving rise to a disaccharide structure composed of a fucosylated GlcNAc, abrogated the affinity of IgG1-Fc for sFcgammaRIIIa. On the other hand, those glycosidase treatments did not significantly affect the affinity of IgG1-Fc for protein A. Inspection of stable-isotope-assisted NMR data of a series of Fc glycoforms indicates that the stepwise trimming out of the carbohydrate residues results in concomitant increase in the number of amino acid residues perturbed thereby in the C(H)2 domains. Furthermore, the cleavage at the GlcNAcbeta1-4GlcNAc glycosidic linkage induced the conformational alterations of part of the lower hinge region, which makes no direct contact with the carbohydrate moieties and forms the major FcgammaR-binding site, while the conformation of the C(H)2/C(H)3 interface was barely perturbed that is the protein A-binding site. These results indicate that the carbohydrate moieties are required for maintaining the structural integrity of the FcgammaR-binding site. PMID- 16343776 TI - Purification and characterisation of a novel enantioselective epoxide hydrolase from Aspergillus niger M200. AB - Purification of a novel enantioselective epoxide hydrolase from Aspergillus niger M200 has been achieved using ammonium sulphate precipitation, ionic exchange, hydrophobic interaction, and size-exclusion chromatography, in conjunction with two additional chromatographic steps employing hydroxylapatite, and Mimetic Green. The enzyme was purified 186-fold with a yield of 15%. The apparent molecular mass of the enzyme was determined to be 77 kDa under native conditions and 40 kDa under denaturing conditions, implying a dimeric structure of the native enzyme. The isoelectric point of the enzyme was estimated to be 4.0 by isoelectric focusing electrophoresis. The enzyme has a broad substrate specificity with highest specificities towards tert-butyl glycidyl ether, para nitrostyrene oxide, benzyl glycidyl ether, and styrene oxide. Enantiomeric ratios of 30 to more than 100 were determined for the hydrolysis reactions of 4 epoxidic substrates using the purified enzyme at a reaction temperature of 10 degrees C. Product inhibition studies suggest that the enzyme is able to differentiate to a high degree between the (R)-diol and (S)-diol product of the hydrolysis reaction with tert-butyl glycidyl ether as the substrate. The highest activity of the enzyme was at 42 degrees C and a pH of 6.8. Six peptide sequences, which were obtained by cleavage of the purified enzyme with trypsin and mass spectrometry analysis of the tryptic peptides, show high similarity with corresponding sequences originated from the epoxide hydrolase from Aspergillus niger LCP 521. PMID- 16343777 TI - Factors controlling neural activity during delayed-response task performance: testing a memory organization hypothesis of prefrontal function. AB - Understanding the role of prefrontal cortex in delayed-response task performance has been a central focus of neuroimaging research. The first part of this review will emphasize consistent observations of memory-load-related effects on prefrontal cortex activity that have led me and my colleagues to propose a "memory-organization hypothesis" of prefrontal cortex function. The second part examines how predictions of this hypothesis have borne up to empirical testing. The final part of this review suggests that there is important information contained in between-study variance in the anatomical locus and temporal sequence of neural activity. I will examine how subtle variations in task-structure affect subjects' strategies, producing meaningful variability in neuroimaging data. Systematic manipulation of these variables in future research can assist in elucidating the role of prefrontal cortex in delayed response task performance. PMID- 16343778 TI - AT4 receptor activation increases intracellular calcium influx and induces a non N-methyl-D-aspartate dependent form of long-term potentiation. AB - The angiotensin 4 receptor (AT4) subtype is heavily distributed in the dentate gyrus and CA1-CA3 subfields of the hippocampus. Neuronal pathways connecting these subfields are believed to be activated during learning and memory processing. ur laboratory previously demonstrated that application of the AT4 agonist, Norleucine1-angiotensin IV, enhanced baseline synaptic transmission and long-term potentiation, whereas perfusion with the AT4 antagonist, Norleucine1 Leu3-psi(CH2-NH2)3-4-angiotensin IV disrupted long-term potentiation stabilization in area CA1. The objective of the present study was to identify the mechanism(s) responsible for Norleucine1-angiotensin IV-induced increase in hippocampal long-term potentiation. Hippocampal slices perfused with Norleucine1 angiotensin IV for 20 min revealed a notable increase in baseline responses in a non-reversible manner and were blocked by the alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione disodium salt. Infusions of Norleucine1-angiotensin IV prior to, but not after theta burst stimulation, significantly enhanced long-term potentiation compared with control slices. Further, N-methyl-D-aspartate receptor-independent long-term potentiation could be induced by tetanization during the perfusion of Norleucine1 angiotensin IV in the presence of the N-methyl-D-aspartate antagonist, D,L-2 amino-5-phosphonovaleric acid. Blockade of select voltage dependent calcium channels significantly reduced Norleucine1-angiotensin IV-induced increase in baseline responses and subsequent long-term potentiation suggesting that AT4 receptor activation increases intracellular calcium levels via altering voltage dependent calcium channels and triggers an N-methyl-D-aspartate-independent form of long-term potentiation. In support of this notion the application of Nle1 angiotensin IV to cultured rat hippocampal neurons resulted in increased intracellular calcium derived exclusively from extracellular sources. Consistent with these observations Nle1-angiotensin IV was capable of augmenting the uptake of 45Ca2+ into rat hippocampal slices. Taken together, these data indicate that increased calcium influx through postsynaptic calcium channels contribute to Norleucine1-angiotensin IV-induced enhancement of long-term potentiation. PMID- 16343779 TI - Evidence for hierarchical error processing in the human brain. AB - Human goal-directed behavior depends on multiple neural systems that monitor and correct for different types of errors. For example, tracking errors in continuous motor tasks appear to be processed by a system involving posterior parietal cortex, whereas errors in speeded response and trial-and-error learning tasks appear to be processed by a system involving frontal-medial cortex. To date, it is unknown whether there is a functional relationship between the posterior and frontal error systems. We recorded the event-related brain potential from participants engaged in a tracking task to investigate the role of the frontal system in continuous motor control. Our results demonstrate that tracking errors elicit temporally distinct error-related event-related brain potentials over frontal and posterior regions of the scalp, suggesting an interaction between the subcomponents of a hierarchically organized system for error processing. Specifically, we propose that the frontal error system assesses high-level errors (i.e. goal attainment) whereas the posterior error system is responsible for evaluating low-level errors (i.e. trajectory deviations during motor control). PMID- 16343780 TI - Latent inhibition of conditioned taste aversion is not disrupted, but can be enhanced, by selective nucleus accumbens shell lesions in rats. AB - Latent inhibition is a form of negative priming in which repeated non-reinforced pre-exposures to a stimulus retard subsequent learning about the predictive significance of that stimulus. The nucleus accumbens shell and the anatomical projection it receives from the hippocampal formation have been attributed a pivotal role in the control or regulation of latent inhibition expression. A number of studies in rats have demonstrated the efficacy of selective shell lesions to disrupt latent inhibition in different associative learning paradigms, including conditioned active avoidance and conditioned emotional response. Here, we extended the test to the conditioned taste aversion paradigm, in which the effect of direct hippocampal damage on latent inhibition remains controversial. We demonstrated the expected effect of selective shell lesions on latent inhibition of conditioned emotional response and of conditioned active avoidance, before evaluating in a separate cohort of rats the effect of comparable selective lesions on latent inhibition of conditioned taste aversion: a null effect of the lesions was first obtained using parameters known to be sensitive to amphetamine treatment, then an enhancement of latent inhibition was revealed with a modified conditioned taste aversion procedure. Our results show that depending on the associative learning paradigm chosen, shell lesions can disrupt or enhance the expression of latent inhibition; and the pattern is reminiscent of that seen following hippocampal damage. PMID- 16343781 TI - GABAB heterodimeric receptors promote Ca2+ influx via store-operated channels in rat cortical neurons and transfected Chinese hamster ovary cells. AB - The GABAB receptors are generally considered to be classical Gi-coupled receptors that lack the ability to mobilize intracellular Ca2+ without the aid of promiscuous G proteins. Here, we report the ability of GABAB receptors to promote calcium influx into primary cultures of rat cortical neurons and transfected Chinese hamster ovary cells. Chinese hamster ovary cells were transfected with GABAB1(a) or GABAB1(b) subunits along with GABAB2 subunits. In experiments using the fluorometric imaging plate reader platform, GABA and selective agonists promoted increases in intracellular Ca2+ levels in transfected Chinese hamster ovary cells and cortical neurons with the expected order of potency. These effects were fully antagonized by selective GABAB receptor antagonists. To investigate the intracellular pathways responsible for mediating these effects we employed several pharmacological inhibitors. Pertussis toxin abolished GABAB mediated Ca2+ increases, as did the phospholipase Cbeta inhibitor U73122. Inhibitor 2-aminethoxydiphenyl borane acts as an antagonist at inositol 1,4,5 trisphosphate receptors and at store-operated channels. In all cell types, 2 aminethoxydiphenyl borane prevented Ca2+ mobilization. The selective store operated channel inhibitor 1-[2-(4-methoxyphenyl)-2-[3-(4 methoxyphenyl)propoxy]ethyl-1H-imidazole hydrochloride prevented increases in intracellular Ca2+ levels as did performing the assays in Ca2+ free buffers. In conclusion, GABAB receptors expressed in Chinese hamster ovary cells and endogenously expressed in rat cortical neurons promote Ca2+ entry into the cell via the activation of store-operated channels, using a mechanism that is dependent on Gi/o heterotrimeric proteins and phospholipase Cbeta. These findings suggest that the neuronal effects mediated by GABAB receptors may, in part, rely on the receptor's ability to promote Ca2+ influx. PMID- 16343782 TI - Identification of adrenoceptor subtype-mediated changes in the density of synapses in the rat visual cortex. AB - Both serotonin and noradrenaline affect synapse formation and maintenance in the CNS. Although we previously demonstrated that serotonin regulates synaptic density via activation of serotonin(2A) receptor, it was still unclear which receptor subtype mediates the function of noradrenaline. In the present study we tried to identify the noradrenaline receptor (adrenoceptor) subtype, which could regulate the density of synapses in the rat visual cortex. Selective antagonists and/or agonists of adrenoceptor subtypes were administered to six weeks old rats. Changes in the density of axodendritic synapses were quantitatively examined in lamina I, where noradrenaline rather than serotonin is known to regulate the density of synapses. The alpha1 adrenoceptor antagonists (prazosin and 2-{[b-(4 hydroxyphenyl)ethyl]aminomethyl}-1-tetralone) decreased the number of synapses in a dose-dependent manner. In contrast, administrations of the alpha1-agonist (methoxamine) increased the density of synapses. The beta1 adrenoceptor antagonist (atenolol) had no effect on the density of synapses. The alpha2 antagonist (rauwolscine) increased synaptic density, whereas the beta2-antagonist (ICI-118,551) decreased synaptic density. Simultaneous treatments with the alpha1 antagonist and alpha1-agonist caused the alpha1-agonist to competitively block the effect of the alpha1-antagonist and recover the density of synapses to the control values. In addition, the alpha1-antagonist/agonist appeared to show a reverse effect on the changes in synaptic density following alpha2- or beta2 antagonist treatment by acting via the alpha1 receptor. Moreover, decreased synaptic density when a selective noradrenergic neurotoxin (N-(2-chloroethyl)-N ethyl-2-bromobenzylamine) was counterbalanced by the alpha1-agonist. These data suggest that noradrenaline regulates the density of synapses in the rat visual cortex primarily via the alpha1 receptor subtype. Both serotonin(2A) and alpha1 receptors are known to couple with phospholipase C, which has been shown to increase intracellular calcium. It may help us to understand the underlying mechanisms for synaptic plasticity in the CNS. PMID- 16343783 TI - From structural biochemistry to expression profiling: neuroprotective activities of estrogen. AB - Estrogens are neuromodulatory and neuroprotective hormones. Chemically, estrogens are steroid compounds and unfold most of their activities through the activation of nuclear receptors that bind to specific target genes and control their transcription. Two subtypes of estrogen receptors are known (estrogen receptor alpha and estrogen receptor beta) and they are expressed throughout the body including the CNS and in particular the brain. We employed large scale DNA-chip analysis to display the gene expression pattern differentially regulated by both estrogen receptor subtypes in human neuronal cells. We identified different gene families regulated by estrogen receptors that complement the knowledge about the estrogen receptor target genes. Some of these genes may serve neuroprotective functions and may therefore mediate the overall neuroprotective activities of estrogens. In addition to estrogen receptor-dependent neuroprotective effects, estrogen (17beta-estradiol) itself is a compound with a phenolic structure that may display also direct and estrogen receptor-independent antioxidant activities which may be important for the defense against oxidative stress. In summary estrogen can display a wide range of neuroprotective activities through different types of mechanisms and we are only understanding part of the molecular control of these activities which may help to develop neuropreventive strategies against neurodegenerative diseases in the future. PMID- 16343784 TI - The putative neural stem cell marker, nestin, is expressed in heterogeneous cell types in the adult rat neocortex. AB - Nestin is a putative neural stem cell marker that is expressed in different areas of the adult mammalian brain that are known to support mitotic activity. Recently the neocortex has been proposed to support neurogenesis, however little is known of the expression pattern of nestin in neocortex. In the present study, we demonstrate that cells that express nestin can be found throughout the neocortex, and that these cells are morphologically heterogeneous. Some nestin-expressing cells had one extension arising from the cell body, reminiscent of the nestin expressing cells that are thought to be young neurons in the hippocampus. The frequency of single extension cells in the neocortex was approximately one cell per 50,000 microm(2). Other cells had numerous extensions arising from the cell body. In all cases, cells that expressed nestin were also found to co-label with the glial marker glial fibrillary acidic protein. In addition, nestin-expressing cells in the neocortex did not express the cell cycle marker, Ki-67, indicating they were not actively engaged in mitotic activity. When small lesions were made in cortex, nestin could also be observed in reactive astrocytes as part of the inflammatory response. Approximately 94% of reactive astrocytes expressed Ki-67. These results demonstrate that there are different populations of cells in the neocortex that can express nestin, but that only reactive astrocytes in this region are mitotically active. PMID- 16343785 TI - Working memory for visual objects: complementary roles of inferior temporal, medial temporal, and prefrontal cortex. AB - Humans have an extraordinary ability to maintain and manipulate visual image information in the absence of perceptual stimulation. The neural substrates of visual working memory have been extensively researched, but there have been few attempts to integrate these findings into a model of how different cortical areas interact to form and maintain visual memories. In this paper, I review findings from neurophysiological, neuropsychological, and neuroimaging studies of visual working memory in human and nonhuman primates. These data support a model in which visual working memory operations rely on activation of object representations in inferior temporal cortex, via top-down feedback from neocortical areas in the prefrontal and medial temporal cortex, and also from the hippocampus. PMID- 16343786 TI - Neuroactive steroids: old players in a new game. AB - It is now clear that the study of the effects exerted by steroids on the nervous system may be considered as one of the most interesting and promising topics for biomedical research. Indeed, new effects, mechanisms of action and targets are becoming more and more evident suggesting that steroids are not only important key regulators of nervous system function but they may also represent a new therapeutic tool to combat certain diseases of the nervous system. The present review summarizes recent observations on this topic indicating that while the concept of the nervous system as a target for steroid hormones has been appreciated for decades, a promising new era for the study of these molecules and their actions in the nervous system has been initiated in the last few years. PMID- 16343787 TI - Adenosine A1 receptors decrease thalamic excitation of inhibitory and excitatory neurons in the barrel cortex. AB - Caffeine is consumed worldwide to enhance wakefulness, but the cellular mechanisms are poorly understood. Caffeine blocks adenosine receptors suggesting that adenosine decreases cortical arousal. Given the widespread innervation of the cerebral cortex by thalamic fibers, adenosine receptors on thalamocortical terminals could provide an efficient method of limiting thalamic activation of the cortex. Using a mouse thalamocortical slice preparation and whole-cell patch clamp recordings, we examined whether thalamocortical terminals are modulated by adenosine receptors. Bath application of adenosine decreased excitatory postsynaptic currents elicited by stimulation of the ventrobasal thalamus. Thalamocortical synapses onto inhibitory and excitatory neurons were equally affected by adenosine. Adenosine also increased the paired pulse ratio and the coefficient of variation of the excitatory postsynaptic currents, suggesting that adenosine decreased glutamate release. The inhibition produced by adenosine was reversed by a selective antagonist of adenosine A1 receptors (8 cyclopentyltheophylline) and mimicked by a selective A1 receptor agonist (N6 cyclopentyladenosine). Our results indicate that thalamocortical excitation is regulated by presynaptic adenosine A1 receptors and provide a mechanism by which increased adenosine levels can directly reduce cortical excitability. PMID- 16343788 TI - Investigating principles of human brain function underlying working memory: what insights from schizophrenia? AB - Working memory dysfunction is a core component of schizophrenia, which likely contributes substantially to the pervasive and profound cognitive deficits observed in patients with this illness. Developments in functional imaging have facilitated the investigation of the neural basis of these cognitive deficits. A strong tradition within neuropsychology has been that circumscribed lesions provide observations which constrain theoretical models, and generate testable predictions on the basis of observed relationships between structural abnormalities and behavioral dysfunction. In this article, the extent to which the neuropsychological tradition can be applied to neuropsychiatry to advance understanding of the biological basis of working memory is addressed. Empirical studies in schizophrenia research are reviewed in relation to principles of normal brain function sub-serving working memory: the functional role of the lateral prefrontal cortex, physiological response capacity constraints, inter regional functional integration, and compensatory adaptations. However, complex heterogeneous psychiatric disorders such as schizophrenia cannot be considered akin to a pure lesion model, and there are considerable methodological challenges in interpreting disruptions of working memory in psychiatric conditions, resulting from clinical, treatment and performance related confounds. The increasing use of psychopharmacological models of disease in healthy human subjects is therefore considered as an attempt to address, or to some extent circumvent these issues. PMID- 16343789 TI - Highly purified lipoteichoic acid from gram-positive bacteria induces in vitro blood-brain barrier disruption through glia activation: role of pro-inflammatory cytokines and nitric oxide. AB - The co-culture of bovine brain capillary endothelial cells and rat primary glial cells was established as an in vitro blood-brain barrier model to investigate the mechanisms by which the Gram-positive bacterial cell wall components lipoteichoic acid and muramyl dipeptide induced injury of blood-brain barrier structure and function. We found that highly purified lipoteichoic acid disrupted blood-brain barrier integrity in a concentration- and time-dependent manner indirectly, through glia activation. Low trans-endothelial electrical resistance and high permeability to fluorescein isothiocyanate-inulin observed in the presence of lipoteichoic acid-activated glial cells were potentiated by muramyl dipeptide and could be reversed only when glial cells were activated by lipoteichoic acid at 10 microg/ml but not with a higher lipoteichoic acid concentration (30 microg/ml). Immunocytochemistry analysis revealed no evident changes in the distribution of the cytoskeleton protein F-actin and tight junction proteins occludin and claudin after lipoteichoic acid treatment. However, the tight junction associated protein AHNAK clearly revealed the morphological alteration of the endothelial cells induced by lipoteichoic acid. Lipoteichoic acid-activated glial cells produced nitric oxide and pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-1beta) that contributed to lipoteichoic acid-induced blood-brain barrier disruption, since the direct treatment of the endothelial monolayer with tumor necrosis factor-alpha or interleukin-1beta increased blood-brain barrier permeability, whereas the pre-treatment of lipoteichoic acid-activated glial cells with antibodies against these two cytokines blocked lipoteichoic acid effects. Additionally, nitric oxide was also involved in blood-brain barrier damage, since the nitric oxide donor itself (diethylenetriamine-nitric oxide adduct) increased blood-brain barrier permeability and inducible nitric oxide synthase inhibitor (1400W) partially reversed lipoteichoic acid-induced trans endothelial electrical resistance decrease. PMID- 16343790 TI - Estrogen activates rapid signaling in the brain: role of estrogen receptor alpha and estrogen receptor beta in neurons and glia. AB - The aging process is known to coincide with a decline in circulating sex hormone levels in both men and women. Due to an increase in the average lifespan, a growing number of post-menopausal women are now receiving hormone therapy for extended periods of time. Recent findings of the Women's Health Initiative, however, have called into question the benefits of long-term hormone therapy for treating symptoms of menopause. The results of this study are still being evaluated, but it is clear that a better understanding of the molecular effects of estradiol is needed in order to develop new estrogenic compounds that activate specific mechanisms but lack adverse side effects. Traditionally, the effects of estradiol treatment have been ascribed to changes in gene expression, namely transcription at estrogen response elements. This review focuses on emerging information that estradiol can also activate a repertoire of membrane-initiated signaling pathways and that these rapid signaling events lead to functional changes at the cellular level. The various types of cells in the brain can respond differently to estradiol treatment based on the signaling properties of the cell, as well as which receptor, estrogen receptor alpha and/or estrogen receptor beta, is expressed. Taken together, these findings suggest that the estradiol-induced activation of membrane-initiated signaling pathways occurs in a cell-type specific manner and can differentially influence how the cells respond to various insults. PMID- 16343791 TI - Grouping of brain rhythms in corticothalamic systems. AB - Different brain rhythms, with both low-frequency and fast-frequency, are grouped within complex wave-sequences. Instead of dissecting various frequency bands of the major oscillations that characterize the brain electrical activity during states of vigilance, it is conceptually more rewarding to analyze their coalescence, which is due to neuronal interactions in corticothalamic systems. This concept of unified brain rhythms does not only include low-frequency sleep oscillations but also fast (beta and gamma) activities that are not exclusively confined to brain-activated states, since they also occur during slow-wave sleep. The major factor behind this coalescence is the cortically generated slow oscillation that, through corticocortical and corticothalamic drives, is effective in grouping other brain rhythms. The experimental evidence for unified oscillations derived from simultaneous intracellular recordings of cortical and thalamic neurons in vivo, while recent studies in humans using global methods provided congruent results of grouping different types of slow and fast oscillatory activities. Far from being epiphenomena, spontaneous brain rhythms have an important role in synaptic plasticity. The role of slow-wave sleep oscillation in consolidating memory traces acquired during wakefulness is being explored in both experimental animals and human subjects. Highly synchronized sleep oscillations may develop into seizures that are generated intracortically and lead to inhibition of thalamocortical neurons, via activation of thalamic reticular neurons, which may explain the obliteration of signals from the external world and unconsciousness during some paroxysmal states. PMID- 16343792 TI - Banishing the homunculus: making working memory work. AB - The prefrontal cortex has long been thought to subserve both working memory and "executive" function, but the mechanistic basis of their integrated function has remained poorly understood, often amounting to a homunculus. This paper reviews the progress in our laboratory and others pursuing a long-term research agenda to deconstruct this homunculus by elucidating the precise computational and neural mechanisms underlying these phenomena. We outline six key functional demands underlying working memory, and then describe the current state of our computational model of the prefrontal cortex and associated systems in the basal ganglia (BG). The model, called PBWM (prefrontal cortex, basal ganglia working memory model), relies on actively maintained representations in the prefrontal cortex, which are dynamically updated/gated by the basal ganglia. It is capable of developing human-like performance largely on its own by taking advantage of powerful reinforcement learning mechanisms, based on the midbrain dopaminergic system and its activation via the basal ganglia and amygdala. These learning mechanisms enable the model to learn to control both itself and other brain areas in a strategic, task-appropriate manner. The model can learn challenging working memory tasks, and has been corroborated by several important empirical studies. PMID- 16343793 TI - Sensation seeking and alcohol use: a meta-analytic review. AB - A meta-analysis was conducted to investigate the association between sensation seeking and alcohol use. Analysis of 61 studies revealed a small to moderate size, heterogeneous effect (mean weighted r=.263) between alcohol use and sensation seeking total scale scores. Analysis of the four sensation seeking components indicated that disinhibition was most strongly correlated with alcohol use (mean weighted r=.368). Categorical moderator analyses revealed that studies that did, versus did not, analyze covariates yielded smaller effect sizes. Continuous moderator analyses indicated that earlier published studies and higher percentages of male and white Caucasian participants were associated with stronger effect sizes between sensation seeking and alcohol use. Novel statistical methodologies for analyzing the association between sensation seeking and alcohol use were proposed and recommendations for future research were suggested. PMID- 16343794 TI - The 'Sutured Skull' and intracranial bleeding in infants. AB - It is known that retinal haemorrhages can result in adults when elevated intrathoracic pressures due to coughing, cardiopulmonary resuscitation, etc., force blood into the head. In infants under one year of age retinal and intracranial haemorrhage commonly occur together, but the same is not true for the older child and adult. The role of the elasticity of the infant skull (resulting from suture and fontanelle stretching) compared to the rigid mature skull, was investigated in a computer aided method. This showed that although in the event of Valsalva-like situations very high lumen pressures may be present in both groups, in the rigid adult skull an immediate corresponding increase in intracranial pressure is produced which surrounds and supports vascular walls leaving transmural pressures little changed. No such support is provided in the eye, and retinal vessels may rupture. Within the skull there may be drastic effects on brain circulation, but since changes in vascular transmural pressure are minimal vessel distension is not induced. In the infant skull the sutures stretch as pressure rises. Since vascular volume is only about 5% of intracranial volume each 1% increase in skull volume permits a 20% increase in vascular volume. Quite small skull expansions will allow dangerous vascular distension and risk of wall damage. Until skull bones fuse, intra-cranial bleeding will be expected in the soft infant skull in any situation where retinal haemorrhage alone is known to occur in the adult or child. PMID- 16343795 TI - Mechanism of endogenous anti-epileptogenesis during rapid eye movement sleep. PMID- 16343796 TI - Simvastatin may be a useful therapy in Dupuytren contracture. PMID- 16343797 TI - Atypical lymphoplasmacytic and immunoblastic proliferation from rheumatoid arthritis: a case report. AB - A case of atypical lymphoplasmacytic and immunoblastic proliferation (ALPIBP) in the lymph nodes associated with well-documented rheumatoid arthritis (RA) is presented. A 68-year-old Japanese female with a 6-year history of RA presented with right neck lymphadenopathy of 3 months duration. A biopsy specimen showed paracortical hyperplasia and numerous lymphoid follicles. On high-power field, the paracortical area was diffusely infiltrated by a polymorphous population consisting of numerous mature plasma cells, plasmacytoid cells, immunoblasts, including Hodgkin-like cells, small- to medium-sized lymphocytes, and histiocytes. Immunohistochemical study demonstrated that immunoblasts usually were CD20+, and a portion of them was CD30+. The histomorphological findings of the present case are similar to those of methotrexate (MTX)-induced atypical lymphoproliferative disorders (LPDs) in some aspects. However, Epstein-Barr virus encoded small RNA-positive cells were not identified by in situ hybridization. The polytypic nature of B lymphocytes also was demonstrated by immunohistochemistry and polymerase chain reaction. Moreover, there was no history of MTX therapy in the present case, indicating that MTX-induced, LPD-like ALPIB may occur even in the RA patients not treated with MTX therapy. PMID- 16343798 TI - Hyperglycemia increased brain ischemia injury through extracellular signal regulated protein Kinase. AB - This study was to examine the alterations in the phosphorylation of mitogen activated protein kinase (MAPK) family in transient brain ischemia under a hyperglycemia and to highlight the molecular mechanisms by which hyperglycemia exacerbates brain damage resulting from stroke. Extracellular signal-regulated protein kinase (ERK) expression was studied in rats subjected to global brain ischemia with pre-ischemic normoglycemic (CIN) and hyperglycemic (CIH) conditions. In another group, the hyperglycemic ischemic rats were pretreated with ERK inhibitor U0126 (U0126). Increased phospho-ERK1/2 immunoreactive neurons in the cingulate cortex and hippocampal CA3 were detected in CIN after ischemia and reperfusion. The numbers of phospho-ERK1/2-positive neurons were further increased significantly in CIH compared to the CIN. Pretreatment with U0126 in CIH rats significantly decreased ERK1/2 immunoreactive cells. Western blot analyses confirmed that phospho-ERK1/2 increased significantly after 30 min ischemia and reperfusion compared to non-ischemic controls in both the CIN and CIH groups. The increase of phospho-ERK1/2 was more prominent in the CIH than in the CIN group after 3 and 6h of reperfusion. Treatment with U0126 significantly reduced phospho-ERK1/2 in the CIH group. The findings presented here suggest that ERK1/2 may play a role in mediating neuronal cells death under hyperglycemic condition. PMID- 16343799 TI - Correlative insights into the immunoexpression of transforming growth factor beta 1 in acutely rejected renal allografts. AB - Recent data suggest that early changes in the process of acute renal transplant rejection (ARTR) occurring during the first 3 months after transplantation include interstitial fibrosis. Transforming growth factor beta (TGF-beta) has been recognized as a key mediator of renal fibrogenesis; therefore, the present study was conducted to ascertain immunoexpression of TGF-beta in ARTR. Another purpose of our study was to determine whether TGF-beta could correlate with the interstitial area and to examine a possible relationship between TGF-beta and interstitial alpha-smooth muscle actin (alpha-SMA), endothelin-1 (ET-1) expression, interstitial T lymphocytes, and monocytes/macrophages. Twenty-four renal allograft biopsy specimens obtained from patients with ARTR were examined using percutaneous renal biopsy. As a control, we used 11 allograft biopsy specimens obtained from patients without any sign of rejection. Staining intensities of TGF-beta-1 in tubuli and of ET-1 in the endothelium of peritubular capillaries, in arterioles, and in the renal tubular epithelial cells were recorded semiquantitatively, whereas interstitial CD3+ cells, CD68+ cells, alpha SMA expression, and the interstitial area were assessed quantitatively using computer image analysis system. Our study revealed that in the ARTR group, the mean values of the immunoexpression of TGF-beta-1, ET-1, interstitial CD3+ cells, CD68+ cells, alpha-SMA expression, and the interstitial area were significantly increased as compared with controls. In the ARTR group, there were significant positive correlations between immunostaining of TGF-beta-1 and ET-1, immunostaining of TGF-beta-1 and alpha-SMA, as well as immunostaining of TGF-beta 1 and interstitial volume. The correlation between immunostaining of TGF-beta-1 and CD 3+ cells tended to be negative; however, this did not reach statistical significance. We did not find any significant relationship between TGF-beta-1 and interstitial monocytes/macrophages. In controls, all these correlations were not significant. In conclusion, our correlative study suggests a role of TGF-beta-1 in early interstitial fibrotic changes in acutely rejected renal allografts, and we hypothesize that endothelin-1 and myofibroblasts pathways may play an important role in this process. PMID- 16343800 TI - Perivascular epithelioid cell sarcoma (malignant PEComa) of the ileum. AB - Epithelioid angiomyolipoma (AML) is the prototype of a heterogeneous group of lesions characterized by the presence of HMB-45 positive cells with clear cytoplasm, perivascular distribution, and combined myomelanocytic features, so called perivascular epithelioid cells (PECs). These lesions are being increasingly referred to as PEComas. PEComas have been reported at diverse anatomic sites, but mainly in the abdominopelvic cavity and rarely in parenchymatous organs, skin, and soft tissues. Gastrointestinal (GI) PEComas are exceptionally rare, with less than 10 cases documented so far. Rare examples of PEComas with pleomorphic histology could have been misinterpreted as unusual variants of carcinoma or sarcoma. To make a contribution to the differential diagnosis of difficult-to-classify pleomorphic GI sarcomas, we report on a malignant pleomorphic neoplasm with features of PEComa involving the terminal ileum in a 63-year-old woman. Fourteen months after resection of the primary tumor, a huge abdominopelvic recurrence was successfully resected, but no distant metastases were detected. The differential diagnosis and malignancy criteria of GI PEComas will be discussed. PMID- 16343801 TI - Supratentorial primitive neuroectodermal tumors (S-PNET) in children: A prospective experience with adjuvant intensive chemotherapy and hyperfractionated accelerated radiotherapy. AB - PURPOSE: Supratentorial primitive neuroectodermal tumors (S-PNET) are rare and have a grim prognosis, frequently taking an aggressive course with local relapse and metastatic spread. We report the results of a mono-institutional therapeutic trial. METHODS AND MATERIALS: We enrolled 15 consecutive patients to preradiation chemotherapy (CT) consisting of high-dose methotrexate, high-dose etoposide, high dose cyclophosphamide, and high-dose carboplatin, craniospinal irradiation (CSI) with hyperfractionated accelerated radiotherapy (HART) plus focal boost, maintenance with vincristine/lomustine or consolidation with high-dose thiotepa followed by autologous stem-cell rescue. RESULTS: Median age was 9 years; 7 were male, 8 female. Site of disease was pineal in 3, elsewhere in 12. Six patients were had no evidence of disease after surgery (NED). Of those with evidence of disease after surgery (ED), 2 had central nervous system spread. Of the 9 ED patients, 2 had complete response (CR) and 2 partial response (PR) after CT, 4 stable disease, and 1 progressive disease. Of the 7 ED patients before radiotherapy, 1 had CR, 4 PR, and 2 minor response, thus obtaining a 44% CR + PR after CT and 71% after HART. Because of rapid progression in 2 of the first 5 patients, high-dose thiotepa was systematically adopted after HART in the subsequent 10 patients. Six of 15 patients relapsed (4 locally, 1 locally with dissemination, 1 with dissemination) a mean of 6 months after starting CT, 2 developed second tumors; 5 of 6 relapsers died at a median of 13 months. Three year progression-free survival, event-free survival, and overall survival were 54%, 34%, and 61%, respectively. CONCLUSION: Hyperfractionated accelerated RT was the main tool in obtaining responses in S-PNET; introducing the myeloablative phase improved the prognosis (3/10 vs. 3/5 relapses), though the outcome remained unsatisfactory despite the adoption of this intensive treatment. PMID- 16343802 TI - X-ray volumetric imaging in image-guided radiotherapy: the new standard in on treatment imaging. AB - PURPOSE: X-ray volumetric imaging (XVI) for the first time allows for the on treatment acquisition of three-dimensional (3D) kV cone beam computed tomography (CT) images. Clinical imaging using the Synergy System (Elekta, Crawley, UK) commenced in July 2003. This study evaluated image quality and dose delivered and assessed clinical utility for treatment verification at a range of anatomic sites. METHODS AND MATERIALS: Single XVIs were acquired from 30 patients undergoing radiotherapy for tumors at 10 different anatomic sites. Patients were imaged in their setup position. Radiation doses received were measured using TLDs on the skin surface. The utility of XVI in verifying target volume coverage was qualitatively assessed by experienced clinicians. RESULTS: X-ray volumetric imaging acquisition was completed in the treatment position at all anatomic sites. At sites where a full gantry rotation was not possible, XVIs were reconstructed from projection images acquired from partial rotations. Soft-tissue definition of organ boundaries allowed direct assessment of 3D target volume coverage at all sites. Individual image quality depended on both imaging parameters and patient characteristics. Radiation dose ranged from 0.003 Gy in the head to 0.03 Gy in the pelvis. CONCLUSIONS: On-treatment XVI provided 3D verification images with soft-tissue definition at all anatomic sites at acceptably low radiation doses. This technology sets a new standard in treatment verification and will facilitate novel adaptive radiotherapy techniques. PMID- 16343803 TI - Radiotherapy for solitary extramedullary plasmacytoma in the head-and-neck region: A dose greater than 45 Gy to the target volume improves the local control. AB - PURPOSE: Our aim was to determine the dose to the clinical target volume (CTV) required for solitary extramedullary plasmacytoma (EMP) in the head and neck (HN). METHODS AND MATERIALS: Seventeen patients (15 Stage I and 2 Stage II) were treated for HN EMP at our institution between 1979 and 2003. The mean International Commission on Radiation Units (ICRU) dose prescribed to the CTV was 52.6 Gy (range, 40-65 Gy) over 24 fractions (range: 20-30). The Stage II patients received neck irradiation doses of 40 and 60 Gy. A mean dose of 36.4 Gy was used for 5 Stage I patients who received elective neck irradiation. Dose administrated to the CTV was evaluated from dosimetric data or from planning films when dosimetric data were not available. Two groups of patients were distinguished: CTV covered with a dose greater than 40 Gy and CTV covered with a dose greater than 45 Gy. RESULTS: The 5-year local control was 72.8%. It was 100% for patients who received dose to the CTV > or = [DOSAGE ERROR CORRECTED] 45 Gy vs. 50% for dose to the CTV <45 Gy (p = 0.034). The prognostic factor for 5-year disease specific survival (81.6%) was local control (p = 0.058). The prognostic factors for disease-free survival (64.1%) were monoclonal immunoglobulin secretion (p = 0.008) and a CTV dose > or = 45 Gy (p = 0.056) CONCLUSIONS: Local control of EMP in the HN seems to be improved when the dose to the CTV is > or = [DOSAGE ERROR CORRECTED] 45 Gy. A minimum dose of 45 Gy should be recommended to the CTV. PMID- 16343804 TI - Resistance of hypoxic cells to ionizing radiation is influenced by homologous recombination status. AB - PURPOSE: To determine the role of DNA repair in hypoxic radioresistance. METHODS AND MATERIALS: Chinese hamster cell lines with mutations in homologous recombination (XRCC2, XRCC3, BRAC2, RAD51C) or nonhomologous end-joining (DNA PKcs) genes were irradiated under normoxic (20% oxygen) and hypoxic (<0.1% oxygen) conditions, and the oxygen enhancement ratio (OER) was calculated. In addition, Fanconi anemia fibroblasts (complementation groups C and G) were compared with fibroblasts from nonsyndrome patients. RAD51 foci were studied using immunofluorescence. RESULTS: All hamster cell lines deficient in homologous recombination showed a decrease in OER (1.5-2.0 vs. 2.6-3.0 for wild-types). In contrast, the OER for the DNA-PKcs-deficient line was comparable to wild-type controls. The two Fanconi anemia cell strains also showed a significant reduction in OER. The OER for RAD51 foci formation at late times after irradiation was considerably lower than that for survival in wild-type cells. CONCLUSION: Homologous recombination plays an important role in determining hypoxic cell radiosensitivity. Lower OERs have also been reported in cells deficient in XPF and ERCC1, which, similar to homologous recombination genes, are known to play a role in cross-link repair. Because Fanconi anemia cells are also sensitive to cross-linking agents, this strengthens the notion that the capacity to repair cross-links determines hypoxic radiosensitivity. PMID- 16343805 TI - Early determination of uterine cervical squamous cell carcinoma radioresponse identifies high- and low-response tumors. AB - PURPOSE: To investigate whether early-assessed radioresponse of tumors corresponds with late-assessed radioresponse, which is associated with local disease control in radiotherapy (RT) for cervical cancer. METHODS AND MATERIALS: This prospective study included 12 patients with cervical squamous cell carcinoma treated by RT with or without concurrent cisplatin. Tumor volume was estimated by scheduled magnetic resonance imaging before (preRT), 3 to 4 weeks after (early assessment), and 6 to 7 weeks after (late assessment) RT initiation. Radioresponse was assessed with tumor shrinkage curves based on these volumes. Radioresponse for each tumor was calculated as the slope (day(-1)) of the shrinkage curve by fitting to an exponential equation. RESULTS: Early-assessed radioresponse ranged from 0.001 to 0.106 day(-1) (median, 0.021 day(-1)) and late assessed radioresponse from 0.009 to 0.091 day(-1) (median, 0.021 day(-1)), with no significant difference between them (p = 0.1191). The early-assessed radioresponse correlated with the late-assessed radioresponse (R(2) = 0.714, p = 0.0005). CONCLUSIONS: Correspondence between early- and late-assessed radioresponse in a series of tumors showing a wide range of radioresponse was not particularly close overall. However, early assessment of radioresponsiveness did seem to be useful for characterizing those tumors with high or low radioresponsiveness. PMID- 16343806 TI - Acquisition versus steady state in the time-left experiment. AB - To test some predictions of scalar expectancy theory (SET) for the time-left procedure, we performed one experiment with two conditions. In Condition A, pigeons were exposed to two fixed-interval schedules, a fixed-interval (FI) 30s and an FI 60s, each associated with a distinct key and presented on a separate trial. Subsequently, during test trials, the FI 60-s key was illuminated and then after T = 15, 30 or 45 s the FI 30-s key also was illuminated. The main issue was how choice between the two keys varied with T. Condition B replicated Condition A with different FI parameters and T values. The results showed that (a) contrary to SET's predictions, preference changed reliably with testing, which suggests that learning took place during the test trials; (b) within each test trial, pigeons revealed an almost exclusive preference for one of the keys, and (c) at steady state pigeons behaved in the same way as rats. Because SET could not account for these findings we advanced a new descriptive model of performance for the time-left task. The model fit the data well. PMID- 16343807 TI - Duration ratio discrimination in pigeons: a criterion-setting analysis. AB - Pigeons received trials beginning with a sequence of two colors (blue-->yellow) on the center key of a three-key array. The colors lasted different lengths of time. At the end of the sequence pigeons chose between two keys based on a criterial ratio of the temporal sequence. One choice was reinforced if the time ratio was less than the criterion and the alternate choice was reinforced if the time ratio was greater than the criterion. The criterial ratios (first to second duration) were 1:1, 1.5:1, and 3:1. The same set of intervals was used for the different criterion ratios, producing a balanced distribution of time ratios for the 1.5:1 condition, and unbalanced distributions for the 1:1 and 3:1 conditions. That is, for the 1.5:1 condition half of the duration pairs were less than the criterion and half were greater. However, for the 1:1 and 3:1 conditions, more duration pairs were less than (3:1) or greater than (1:1) the criterion. Accuracy was similar across criterion ratios, but response bias was influenced by the asymmetries of time ratios in the 1:1 and 3:1 conditions. When these asymmetries were controlled, the response biases were reduced or eliminated. These results indicate that pigeons are flexible in establishing a criterion for discriminating duration ratios, unlike humans, who are less flexible and are bound to categorical distinctions in the discrimination of duration ratios. PMID- 16343808 TI - Do primary care physicians' own AUDIT scores predict their use of brief alcohol intervention? A cross-sectional survey. AB - BACKGROUND: To define whether the Alcohol Use Disorders Identification Test (AUDIT) scores of primary care physicians themselves predict their willingness to use brief alcohol intervention. METHODS: Cross-sectional self-administered questionnaire survey to all 3193 physicians providing primary health care in Finland. The response rate was 1909 (59.8%). Odds ratios from multinomial regression analysis were calculated for self-reported frequency (never, occasionally or regularly) of conducting brief interventions by physicians with AUDIT scores of 0-1, 2, 3, 4, 5-7 or >or=8. RESULTS: The prevalence of heavy drinkers based on AUDIT score (>or=8) was 14.5% among all physicians, 7.0% among females and 27.0% among males. Of the respondents 9.4% reported doing brief intervention regularly and 50.0% occasionally. AUDIT scores did not significantly predict either regular or occasional use of brief intervention. Instead, some other independent predictors for more frequent use of brief intervention were found. These included having a specialist licence in general practice or occupational health care and the location of the practice, but not gender or age. CONCLUSIONS: The present results indicate that in general heavy drinking among primary care physicians do not explain the low frequency with which brief intervention is used in primary health care. PMID- 16343809 TI - One year outcomes for heroin dependence: findings from the Australian Treatment Outcome Study (ATOS). AB - AIM: To determine 1 year outcomes for drug use, criminality, psychopathology and injection-related health problems in those entering treatment for heroin dependence in Australia. DESIGN: Longitudinal prospective cohort study. PARTICIPANTS: Seven hundred and forty five individuals entering treatment (methadone/buprenorphine maintenance therapy; detoxification; residential rehabilitation) and 80 heroin users not seeking treatment. SETTING: Sydney, Melbourne and Adelaide, Australia. FINDINGS: A total of 657 individuals were re interviewed at 1 year, 80% of the original sample. There were substantial reductions in heroin and other drug use across all three treatment modalities. The majority of those who had entered treatment were heroin abstinent at 1 year (maintenance therapy 65%, detoxification 52%, residential rehabilitation 63%) compared to 25% of the non-treatment sample. The reduction in heroin use among the treatment samples was paralleled by reductions in poly drug use. There were also substantial reductions in risk-taking, crime and injection-related health problems across all treatment groups, and less marked reductions among the non treatment group. Psychopathology was dramatically reduced among the treatment modalities, while remaining stable among the non-treatment group. Positive outcomes at 1 year were associated with a greater number of cumulative treatment days experienced over the 1 year follow-up period ('treatment dose') and fewer treatment episodes undertaken in that time ('treatment stability'). CONCLUSIONS: At 1 year, there were impressive reductions in drug use, criminality, psychopathology and injection-related health problems following treatment exposure. The positive findings were associated with a greater "dose" of treatment, and with more treatment stability over the follow-up period. PMID- 16343810 TI - Prevalence of injecting drug use and associated risk behavior among regular ecstasy users in Australia. AB - BACKGROUND: The aim of the study was to investigate the prevalence of injecting drug use and associated risk behaviour among a sentinel sample of ecstasy users. METHODS: Cross-sectional surveys were conducted with regular ecstasy users as part of an annual monitoring study of ecstasy and related drug markets in all Australian capital cities. RESULTS: Twenty-three percent of the sample reported having ever injected a drug and 15% reported injecting in the 6 months preceding interview. Independent predictors of lifetime injection were older age, unemployment and having ever been in prison. Completion of secondary school and identifying as heterosexual was associated with a lower likelihood of having ever injected. Participants who had recently injected typically did so infrequently; only 9% reported daily injecting. Methamphetamine was the most commonly injected drug. Prevalence of needle sharing was low (6%), although half (47%) reported sharing other injecting equipment in the preceding 6 months. CONCLUSIONS: Ecstasy users who report having injected a drug at some time appear to be demographically different to ecstasy users who have not injected although neither are they typical of other drug injectors. The current investigation suggests that ongoing monitoring of injecting among regular ecstasy users is warranted. PMID- 16343811 TI - Impaired decision-making in opiate-dependent subjects: effect of pharmacological therapies. AB - Cognitive dysfunction is a major feature of drug addiction. In the present paper, we compared the decision-making ability using the Iowa gambling task of methadone and buprenorphine-maintained individuals to non opiate-dependent drug-free controls. Buprenorphine-maintained individuals performed better than methadone maintained individuals, and not differently than non opiate-dependent controls. In addition, methadone-maintained individuals had more perseverative errors on the Wisconsin card sorting task (WCST) as compared with non opiate-dependent drug free controls whereas buprenorphine-maintained individuals had intermediate scores. Scores on Weschler adult intelligence scale (WAIS-R) were similar for methadone- and buprenorphine-maintained individuals whereas drug-free controls had significantly higher scores. In addition, both opiate-dependent groups performed more poorly than drug-free controls on the Benton visual retention test (BVRT). The results suggest that buprenorphine in contrast to methadone improves decision-making, and thus may be more effective in rehabilitation programs of opiate-dependent subjects and this improvement may be related to its distinct pharmacological action as a k antagonist. PMID- 16343813 TI - Origin and diversification of minisatellites derived from human Alu sequences. AB - We analyze minisatellites derived from Alu fragments corresponding approximately to the first 44 bases of human Alu consensus sequences from different subfamilies. The origin of Alu-derived minisatellites appears to have been mediated by short flanking repeats, as first proposed by Haber and Louis [Haber, J.E., Louis, E.J., 1998. Minisatellite origins in yeast and humans. Genomics 48, 132-135.]. We also present evidence for base substitutions and deletions introduced to minisatellites by gene conversion with partially similar but unrelated flanking regions. Segments flanked by short direct repeats are relatively common in different regions of Alu and other repetitive sequences. Our analysis shows that they can be effectively used in comparative studies of the overall sequence context which may contribute to instability of DNA segments flanked by short direct repeats. PMID- 16343812 TI - Identification of highly specific localized sequence motifs in human ribosomal protein gene promoters. AB - For ribosomal protein (RP) genes the start of transcription is rigidly controlled to maintain the 5'-TOP signal on the messenger RNA. The responsible regulatory mechanism is not yet fully understood. Careful comparative analysis of their proximal promoter sequences reveals common characteristics and thus provides clues to the underlying mechanism. We have extracted the proximal promoters of the 80 human cytosolic ribosomal protein genes together with the orthologous mouse sequences. After annotating the set with transcription factor binding sites based on the available literature, we searched for over-represented sequence motifs. We uncovered a novel motif that is localized at a fixed distance downstream to the transcription start. 31 out of the 80 promoters contain the motif in the same orientation around position +62 (standard deviation 6). A second evolutionary conserved and palindromic motif is found 13 times in the RP promoter set, 9 instances of which are located upstream around position -40. In addition, we see a characteristic profile of the GC-content and of the CpG dinucleotide frequencies. Our results support a model for the transcription of ribosomal protein genes in which the maintenance of the accurate start of transcription is provided by specific transcription factors. Such a factor binds the target DNA at a fixed location relative to the TSS, and possibly interacts directly with the basal transcription machinery. PMID- 16343814 TI - A new strategy to identify novel genes and gene isoforms: Analysis of human chromosomes 15, 21 and 22. AB - We present here a novel methodology for the identification of genome regions potentially spanning one or more protein coding genes. It is based on the detection of clusters of conserved sequence tags whose evolutionary dynamics, based on the observation of an excess bias of synonymous substitutions at nucleotide level and of conservative replacements at protein level, suggests a likely protein coding role. A benchmark test carried out on a 236 Mbp of human mouse syntenic regions from human chromosomes 15, 21 and 22 identified 25 CST clusters potentially containing unannotated genes. A further annotation update of the human genome assembly revealed that 11/25 clusters actually contained a total of 20 validated genes and 10 of the remaining 14 clusters had several experimental evidence in support of the presence of protein coding genes. These findings demonstrate the effectiveness and high prediction reliability of the proposed methodology which could specifically be applied to the annotation of novel genome sequences. PMID- 16343815 TI - Clast4, the murine homologue of human eIF4E-Transporter, is highly expressed in developing oocytes and post-translationally modified at meiotic maturation. AB - In metazoans, translational regulation of a set of maternal mRNAs directs oocyte maturation and early embryogenesis. These transcripts are often kept dormant until their products are spatially and temporally required in development. The interaction between general translation factors (i.e. eIF4E) and their specific interactors influences translation initiation. A search of the protein database for a mouse homologue of the Drosophila Cup protein, a translational repressor during female germ-line development, identified the product of the Clast4 gene. In this report, we show that Clast4 mRNA and protein are highly expressed within the cytoplasm of growing oocytes. The Clast4 protein is stable during this developmental window and post-translationally modified by phosphorylation upon oocyte meiotic maturation. Additionally, we show that Clast4 and eIF4E directly interact by means of a canonical and functional eIF4E-binding motif. Our results suggest that Clast4, similar to Drosophila Cup, may act at the translational level during murine female germ-line development. PMID- 16343817 TI - Wildlife reservoirs for bovine tuberculosis (Mycobacterium bovis) in Canada: strategies for management and research. AB - In Canada, there are two known regional foci where wildlife populations are infected with bovine tuberculosis (Mycobacterium bovis) and considered to be disease reservoirs. Free-ranging populations of wood bison (Bison bison athabascae) in and around Wood Buffalo National Park (WBNP) and wapiti (Cervus elaphus manitobensis) in and around Riding Mountain National Park (RMNP) are infected with bovine tuberculosis. In this paper, we provide an overview of these diseased wild ungulate populations and the complexities of attempting to manage issues relating to bovine tuberculosis in and around protected areas. We do not describe the quantitative science and epidemiological data in detail from these case histories, but instead compare and contrast these two cases from a broader perspective. This is achieved by reviewing the context and process by which a diverse group of stakeholders engage and develop strategies to address the controversial problems that diseased wildlife populations often present. We suggest that understanding the factors that drive the strategic-level management processes is equally important for addressing a wildlife disease problem as the tactical-level issues, such as design and implementation of technically sound field research and management programs. Understanding the experiences within the WBNP and RMNP areas, particularly the strategies that have failed or succeeded, may prove useful to understanding and improving management approaches when wildlife are infected with M. bovis. Applying this understanding is consistent with the principles of adaptive management in which we learn from previous experiences to develop better strategies for the future. PMID- 16343816 TI - Tertiary structure and spectral tuning of UV and violet pigments in vertebrates. AB - Many vertebrate species use ultraviolet (UV) vision for such behaviors as mating, foraging, and communication. UV vision is mediated by UV-sensitive visual pigments, which have the wavelengths of maximal absorption (lambda max) at approximately 360 nm, whereas violet (or blue) vision is mediated by orthologous pigments with lambda max values of 390-440 nm. It is widely believed that amino acids in transmembrane (TM) I-III are solely responsible for the spectral tuning of these SWS1 pigments. Recent molecular analyses of SWS1 pigments, however, show that amino acids in TM IV-VII are also involved in the spectral tuning of these pigments through synergistic interactions with those in TM I-III. Comparisons of the tertiary structures of UV and violet pigments reveal that the distance between the counterion E113 in TM III and amino acid sites 87-93 in TM II is narrower for UV pigments than for violet pigments, which may restrict the access of water molecules to the Schiff base pocket and deprotonate the Schiff base nitrogen. Both mutagenesis analyses of E113Q and quantum chemical calculations strongly suggest that unprotonated Schiff base-linked chromophore is responsible for detecting UV light. PMID- 16343818 TI - A review of tuberculosis science and policy in Great Britain. AB - Bovine tuberculosis is one of the most complex animal health problems that the farming industry in Great Britain faces today. In leading and facilitating the changes to policy required to reverse the long-term upward trend in the disease, Government is heavily reliant on evidence emerging from its wide-ranging bovine tuberculosis research programme. The paper outlines development of policy in Great Britain and its relationship to research findings. PMID- 16343819 TI - Wildlife tuberculosis in South African conservation areas: implications and challenges. AB - Tuberculosis, caused by Mycobacterium bovis, was first diagnosed in African buffalo in South Africa's Kruger National Park in 1990. Over the past 15 years the disease has spread northwards leaving only the most northern buffalo herds unaffected. Evidence suggests that 10 other small and large mammalian species, including large predators, are spillover hosts. Wildlife tuberculosis has also been diagnosed in several adjacent private game reserves and in the Hluhluwe iMfolozi Park, the third largest game reserve in South Africa. The tuberculosis epidemic has a number of implications, for which the full effect of some might only be seen in the long-term. Potential negative long-term effects on the population dynamics of certain social animal species and the direct threat for the survival of endangered species pose particular problems for wildlife conservationists. On the other hand, the risk of spillover infection to neighboring communal cattle raises concerns about human health at the wildlife livestock-human interface, not only along the western boundary of Kruger National Park, but also with regards to the joint development of the Greater Limpopo Transfrontier Conservation Area with Zimbabwe and Mozambique. From an economic point of view, wildlife tuberculosis has resulted in national and international trade restrictions for affected species. The lack of diagnostic tools for most species and the absence of an effective vaccine make it currently impossible to contain and control this disease within an infected free-ranging ecosystem. Veterinary researchers and policy-makers have recognized the need to intensify research on this disease and the need to develop tools for control, initially targeting buffalo and lion. PMID- 16343820 TI - Recombination in the alphaherpesvirus bovine herpesvirus 1. AB - Herpesviruses are DNA viruses characterized by a low rate of nucleotide substitution. Therefore, other mechanisms must be involved to their evolution, like recombination that can be seen as an essential evolutionary driving force of these viruses. Recombination contributes to the long-term evolution of alphaherpesviruses. It acts also to continuously create new alphaherpesvirus strains. We have used bovine herpesvirus 1 to investigate recombination both within DNA concatemers in infected cells and in vitro and in vivo at the end of the lytic cycle. The following results have been obtained: (i) intramolecular recombination occurs at the level of concatemers and gives rise to genomic segment inversions; (ii) intraspecific recombination occurs frequently both in vitro and in vivo; (iii) interspecific recombination is possible and requires two highly genetically related viruses; (iv) only simultaneous or closely separated infections lead to the production of recombinant viruses; (v) recombination between wild-type and glycoprotein defective vaccine virus can produce a glycoprotein defective virus keeping part of the virulence of parental wild-type virus. Recombination, by exchanging genomic segments, may modify the virulence of alphaherpesviruses. It must be carefully assessed for the biosafety of antiviral therapy, alphaherpesvirus-based vectors and live attenuated vaccines. PMID- 16343821 TI - Low oxygen exposure does not cause pulmonary injury in the newborn rat. AB - BACKGROUND: Two recent studies have suggested that low levels of supplemental inspired oxygen may cause lung injury in preterm infants. AIMS: To assess lung injury of newborn rats exposed to 14 days of low-level variation of oxygen. STUDY DESIGN: Four groups were compared with 12 animals per group and 4 lung sections per animal. These were, a control group raised in room air and three groups raised in levels of inspired oxygen fluctuating around the following mean values: group Lo (mean FiO(2) 0.179), group N (mean FiO(2) 0.213), and group Hi (mean FiO(2) 0.247). The degree of oxygen variability was identical for each group. Lungs were inflated at 20 cm H(2)O, fixed and stained with H and E and Millers Elastin. SUBJECTS: Sprague Dawley albino newborn rats. OUTCOME MEASURES: Random alveolar areas were studied and analysed using imaging software to assess total amount of tissue and elastin, number of secondary septa, and mean linear intercept. RESULTS: There were no significant differences between the three experimental oxygen groups and the control group in terms of lung/body weight ratio and the measured markers of lung development. CONCLUSION: We conclude that low-level oxygen supplementation during early lung development does not affect alveolar development in the newborn rat. PMID- 16343822 TI - Necrotizing enterocolitis increases the bone resorption in premature infants. AB - BACKGROUND: Necrotizing enterocolitis is a common neonatal gastrointestinal disease that affects approximately 10% of premature infants less than 1500 g. The average mortality is 20-40% and survivors may present with diarrhea or malabsorption, intestinal strictures and fistulas, feeding abnormalities and failure to thrive. It is not clear whether the higher incidence of this gastrointestinal disease in premature infants contributes to the risk of osteopenia of prematurity. AIM: To examine bone turnover state in premature infants who had a necrotizing enterocolitis attack during postnatal period. STUDY DESIGN AND SUBJECTS: We examine the bone turnover markers in infants with necrotizing enterocolitis and compare them with infants with sepsis. Forty-one premature infants participated in the study and were divided into three groups. In group I, there were 14 premature infants who developed necrotizing enterocolitis with negative blood culture during their hospitalization. In group II, there were 12 premature infants who developed sepsis during their hospitalization. Age-matched 15 premature infants who were given parenteral nutrition served as control group (group III). Blood samples and 6-h urine samples were obtained for bone turnover markers and calcium, phosphorous, creatinine and 25-hydroxy vitamin D between the day 20 and 25. Bone osteoblastic activity was assessed by measurement of serum osteocalcin. Bone resorption was assessed by measurement of serum levels of beta-CrossLaps and urinary deoxypyridinoline. RESULTS: There were no significant differences in bone osteoblastic activity among the groups, but bone resorption markers were significantly higher in infants with necrotizing enterocolitis compared to other groups (p < 0.016). CONCLUSION: Necrotizing enterocolitis increases the bone resorption in premature infants. It may be related with reduced glucagon like peptide-2 levels, a new intestinal hormone that is primary secreted from distal small intestine. PMID- 16343823 TI - The hepatic cytochrome P450 reductase null mouse as a tool to identify a successful candidate entity. AB - Cytochrome P450s (CYP) play a pivotal role in the metabolism of drugs and xenobiotics, and have been intensively studied over many years. Much of the work carried out on the role of hepatic cytochrome P450s in drug metabolism and disposition has been done in vitro, and has yielded vital information on P450 regulation and function. However, additional factors such as route of administration, absorption, drug transporters, renal clearance and extra-hepatic P450s, make it difficult to extrapolate from in vitro data to in vivo pharmacokinetics. A number of cytochrome P450s knockout mice have been generated, although many have been of limited usefulness due to either embryonic/perinatal lethality, or the functional redundancy inevitably found in a large family of isoenzymes. We have developed a mouse line (HRN) in which cytochrome P450 oxidoreductase (POR), the unique electron donor to cytochrome P450s is deleted specifically in the liver, resulting in the loss of essentially all hepatic P450 function. The HRN mouse, although having disturbances in lipid and bile acid homeostasis develops and breeds normally. We have used the HRN mouse as a model to establish the role of hepatic versus extra-hepatic metabolism in drug metabolism and disposition, and also to investigate the relationship between drug toxicokinetics and therapeutic effect, initially with the chemotherapeutic prodrug cyclophosphamide (CPA). PMID- 16343824 TI - Endocytosis and intracellular trafficking of fatty acid esters of phenylaminopropanediol, the putative etiologic agents of the toxic oil syndrome (TOS). AB - The toxic oil syndrome (TOS) caused by ingestion of rapeseed oil adulterated with aniline is characterized by symptoms of an allergic and/or autoimmune illness associated with vessel wall lesions similar to those of atherosclerosis. Fatty acid esters of 3-(N-phenylamino)-1,2-propanediol (PAP) have been incriminated as the etiologic agents of TOS. However, the pathogenesis of TOS is yet unknown. Here, we addressed whether PAP fatty acid esters are incorporated into lipoproteins, which after transport to vascular endothelial cells are taken up to initiate TOS vasculopathy. After loading (14)C-dioleyl-ester of PAP into LDL labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindolcarbocyanine (DiI) we assessed receptor mediated endocytosis and intracellular localization of these lipopoproteins in vascular endothelial cells. Our data suggest that these lipoprotein-derivatives are internalized into endothelial cells by LDL receptor mediated endocytosis. Confocal microscopy revealed that DiI-LDL loaded with dioleyl-ester of PAP and incubated for 60 min with endothelial cells colocalizes with the lysosomotropic compound LysoTracker Green, indicating that internalized PAP-loaded LDL are targetted to the endolysosomal compartment for further processing. Subcellular fractionation of endothelial-like ECV-304 cells after incubation with LDL loaded with the (14)C-dioleyl-ester of PAP for 6h showed that the radioactive label accumulated in fractions containing endosomes, the Golgi apparatus and the endoplasmic reticulum. PMID- 16343825 TI - Effect of transport and storage temperature of ovaries on in vitro maturation of bitch oocytes. AB - In this study, the effects of ovary transport and storage temperature on in vitro maturation of bitch oocytes were investigated. Ovaries were collected from 23 mature bitches and one randomly selected ovary of each pair (n=23 pairs) was transported in physiologic saline at 4 degrees C, while the other one at 35-38 degrees C for 2-4h. A total of 316 cumulus oocyte complexes (COCs) were obtained from the 4 degrees C group and 301 COCs from the 35-38 degrees C group. All COCs were matured in modified synthetic oviduct fluid (mSOF) supplemented with follicle stimulating hormone (FSH), essential and non-essential amino acids at 38 degrees C in a humidified 5% CO2, 5% O2, and 90% N2 atmosphere for 72 h. At the end of the in vitro maturation period, nuclear maturation of oocytes was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), metaphase II (MII), undetermined nuclear maturation (UDNM), and MI+MII. The nuclear maturation rates to MI, MII, and MI+MII stages were 60.44%, 10.75%, and 71.20% in the 4 degrees C group and 37.20%, 7.64%, and 45.85% in the 35-38 degrees C group, respectively. The data demonstrated that oocytes obtained from ovaries transported at 4 degrees C had higher maturation rates than from the ones transported at 35-38 degrees C (p<0.001). PMID- 16343826 TI - Effect of suckling on basal and GnRH-induced LH release in post-partum dairy buffaloes. AB - Suckling, a common practice in smallholder dairy-farming systems in the developing world, delays the onset of post-partum ovarian activity in dairy buffalo. The present study was designed to assess the effect of suckling on pituitary function in lactating buffaloes 25-35 days post-partum. Six suckled and nine non-suckled buffaloes were challenged intravenously with a bolus injection of GnRH (20microg buserelin acetate; Receptal). Heparinized venous blood samples were collected at 15min intervals for 2h before and up to 4h after GnRH for luteinizing hormone (LH) estimation. Pretreatment basal LH concentrations were similar in the suckled (0.6+/-0.2ng/ml) and the non-suckled (0.5+/-0.1ng/ml) buffaloes. All but one suckled buffaloes released a LH surge, starting 15-60min post-GnRH treatment, which lasted for 180-225min. While one suckled buffalo did not respond to GnRH, the LH response in the remaining suckled buffaloes was significantly less than in the non-suckled buffaloes in terms of peak LH concentrations (14.3+/-2.7ng/ml versus 26.2+/-4.3ng/ml) and area under the LH curve (1575.6+/-197.4mm(2) versus 2108.9+/-323.9mm(2)). The LH response was least in suckled buffaloes challenged with GnRH while in the luteal phase of an oestrus cycle and with plasma progesterone concentration >1ng/ml. In conclusion, suckling suppressed pituitary responsiveness to exogenous GnRH challenge in post-partum buffaloes. PMID- 16343827 TI - TNFRSF11B gene variants and bone mineral density in postmenopausal women in Malta. AB - A number of polymorphisms in various genes have been identified and associated with bone mineral density (BMD) and with an increased risk of osteoporosis. OBJECTIVE: In this study, three single nucleotide polymorphisms (SNPs) within the TNFRSF11B gene were studied for association with an increased risk of osteoporosis in postmenopausal Maltese women (n=126). METHODOLOGY: Analysis was performed by PCR restriction fragment length polymorphism (RFLP) while BMD at the lumbar spine, femoral neck, Ward's triangle and trochanter was measured by DEXA. RESULTS: No significant association was observed between genotypes and BMD for all polymorphisms studied within this gene. Homozygotes CC (T(950)-C) were observed to have the highest BMD at all anatomical sites although statistical significance was not reached when comparing the three genotypes. A statistical significant difference was observed in the distribution of genotype frequencies for this polymorphism between normal individuals and those that were either osteopenic or osteoporotic at one or both anatomical sites, with the TT genotype associated more frequently with low BMD. The T(950)-C and G(1181)-C polymorphisms were in strong linkage disequilibrium with each other but not with the A(163)-G polymorphism further upstream in the OPG promoter. Statistical significance was reached when constructing haplotypes, where the A-T-G haplotype was found to be more frequent in individuals with low BMD. CONCLUSIONS: These results indicate the possible role of TNFRSF11B gene variants in postmenopausal bone loss in women in Malta. PMID- 16343828 TI - Effects of estrogens on cognition and brain morphology: involvement of the cerebellum. AB - OBJECTIVES: Sex steroid hormones are implicated in the cognitive processes of the adult brain. Among studies reporting a positive effect of estrogen replacement therapy (ERT) on cognition, the most consistent evidence is that it enhances verbal memory and visuospatial functions. In the present study we investigated the effect of ERT on cognition and on brain morphology in healthy postmenopausal women, taking into account the distinction in current and past ERT users. METHODS: Participants were postmenopausal nondemented women recruited from the community: ERT users were 40 (23 current users, 17 past users), while never users were 43. Forty of recruited subjects gave consent to undergo 3D high resolution MRI (16 current users, 7 past users and 17 never users). Participants underwent MMSE and a battery of neuropsychological tests measuring memory, language, intelligence, attention and visuo-spatial abilities. RESULTS: The past users group outperformed the never users in four tests: Token test, WCST categories, attentional matrices and Rey's delayed list; the current users group outperformed the never users in the Rey's list test. ERT users had greater grey matter volumes mainly in the cerebellum, but an increase was observed also in the parietal and occipital cortex. CONCLUSIONS: ERT use appears to improve linguistic, attentive and planning abilities. Interestingly, the beneficial effects on cognition were detected mainly in the past users subgroup. Here we propose that the trophic effect of estrogens on cerebellum might account for the observed improvement in cognition. PMID- 16343829 TI - Citric acid prolongs the gastro-retention of a floating dosage form and increases bioavailability of riboflavin in the fasted state. AB - A floating dosage form based on calcium alginate beads has been developed. Riboflavin, was selected as the model drug and successfully incorporated into calcium alginate beads. The aims of the current study were to: (a) assess the influence of prolonged gastro-retention on the bioavailability of riboflavin from freeze dried calcium alginate beads administered under varying conditions of food intake and (b) to investigate the potential of citric acid to delay the gastric emptying of the calcium alginate beads. Gamma scintigraphy was selected as the method to monitor the movement of the calcium alginate beads in vivo. Riboflavin concentrations in the urine were analysed by HPLC. Prolonged gastro-retention can be achieved, in the fasted state, when citric acid solution is used as an administering vehicle. However, prolonged gastro-retention is not achieved to the same extent when the gastric emptying times are compared to those obtained in the fed state. The bioavailability of riboflavin improved when calcium alginate beads were administered in the fasted state with citric acid solution, compared to the bioavailability obtained when the calcium alginate beads were administered in the absence of citric acid. PMID- 16343830 TI - The effect of BAPTA and 4AP in scala media on transduction and cochlear gain. AB - We have injected by iontophoresis 4-amino-pyridine, a K+ channel blocker and BAPTA, (a Ca++ chelator), into scala media of the first three turns of the guinea pig cochlea. We measured the reduction in outer hair cell (OHC) receptor current, as indicated by cochlear microphonic measured in scala media evoked by a 207 Hz tone, and compared this with the elevation of the cochlear action potential (CAP) threshold. We found that in the basal turn, for frequencies between 12 and 21 kHz, CAP threshold was elevated by about 30 dB, while in the second turn, at the 3 kHz place, the maximum elevation was 15 dB. In the third turn, iontophoresis of 4AP and BAPTA reduced CM by similar amounts to that in the basal and second turn, but caused negligible elevation of CAP threshold. We conclude that the gain of the cochlear amplifier is maximal for basal turn frequencies, is halved at 3 kHz, and is reduced to close to one for frequencies below 1 kHz (no active gain). The effect of 4AP and BAPTA on neural threshold and the receptor current represented by CM may be explained by their action on OHC transduction without the involvement of IHCs. PMID- 16343831 TI - Anti-inflammatory activity of Bacopa monniera in rodents. AB - The ethanol extract of Bacopa monniera (Scrophulariaceae) exhibited marked anti inflammatory activity against carrageenan-induced paw edema in mice and rats, an acute inflammatory model. To assess the possible mechanism of anti-inflammatory action against carrageenan, the ethanol extract was treated with chemical mediators (histamine, serotonin, bradykinin, prostaglandin E(2) and arachidonic acid)-induced edema in rats. The extract selectively inhibited prostaglandin E(2) induced inflammation. Thus, it may be inferred that B. monniera possesses significant anti-inflammatory activity that may well be relevant for its effectiveness in the healing of various inflammatory conditions in traditional medicine. PMID- 16343832 TI - Iatrogenically-related, fatal methotrexate intoxication: a series of four cases. AB - Since the early eighties, the folic antagonist methotrexate (MTX) has been used in long-term treatment of rheumatoid arthritis. Because of the high toxic potential clinical and laboratory controls at regular intervals and patient education in order to avoid misadventure is of overriding importance. We present four cases of fatal MTX intoxication due to medical malpractice from the Tubingen Institute of Forensic Medicine autopsy material, which show the severe consequences of MTX overdose. It becomes evident that among non-rheumatologists there still is need for information about toxicity and dose limitation in MTX low dose treatment. PMID- 16343833 TI - Phylogenetic relationship of psychoactive fungi based on rRNA gene for a large subunit and their identification using the TaqMan assay (II). AB - "Magic mushroom (MM)" is the name most commonly given to psychoactive fungi containing the hallucinogenic components: psilocin (1) and psilocybin (2). We investigated the rRNA gene (internal transcribed spacer (ITS) and large subunit (LSU)) of two Panaeolus species and four Psilocybe species fungi (of these, two are non-psilocybin species). On the basis of sequence alignment, we improved the identification system developed in our previous study. In this paper, we describe the new system capable of distinguishing MMs from non-psilocybin Psilocybe species, its application data and the phylogeny of MM species. PMID- 16343834 TI - Analysis of artificially degraded DNA using STRs and SNPs--results of a collaborative European (EDNAP) exercise. AB - Recently, there has been much debate about what kinds of genetic markers should be implemented as new core loci that constitute national DNA databases. The choices lie between conventional STRs, ranging in size from 100 to 450 bp; mini STRs, with amplicon sizes less than 200 bp; and single nucleotide polymorphisms (SNPs). There is general agreement by the European DNA Profiling Group (EDNAP) and the European Network of Forensic Science Institutes (ENFSI) that the reason to implement new markers is to increase the chance of amplifying highly degraded DNA rather than to increase the discriminating power of the current techniques. A collaborative study between nine European and US laboratories was organised under the auspices of EDNAP. Each laboratory was supplied with a SNP multiplex kit (Foren-SNPs) provided by the Forensic Science Service, two mini-STR kits provided by the National Institute of Standards and Technology (NIST) and a set of degraded DNA stains (blood and saliva). Laboratories tested all three multiplex kits, along with their own existing DNA profiling technique, on the same sets of degraded samples. Results were collated and analysed and, in general, mini-STR systems were shown to be the most effective. Accordingly, the EDNAP and ENFSI working groups have recommended that existing STR loci are reengineered to provide smaller amplicons, and the adoption of three new European core loci has been agreed. PMID- 16343835 TI - Age-associated shifts in cardiac gene transcription and transcriptional responses to ischemic stress. AB - Aged hearts exhibit reduced tolerance to ischemia-reperfusion, together with altered structure and post-ischemic remodelling. The molecular bases of such changes are unclear. Using cDNA microarrays and quantitative RT-PCR we characterized shifts in gene expression patterns with aging in normoxic and post ischemic (20 min global ischemia, 60 min reperfusion) murine hearts (young: 2-4 months; aged: 16-18 months). We identified an age-associated up-regulation of transcripts involved in cell death, oxygen transport and metabolism in normoxic hearts. Down-regulated transcripts were involved in transporter activity, protein binding and hydrolase activity, changes in MAPK, WNT and TGF-beta signalling with aging were also observed. Ischemic stress generated a much greater degree of contractile impairment and cellular damage in aged vs. young hearts. This was associated with a substantially modified transcriptional response, with selective changes in Ca2+, WNT, NOTCH and G-protein coupled receptor signalling paths in aged vs. young hearts. Despite some common responses to ischemia in young and aged hearts (induction of heat shock protein transcripts), aging selectively modified ischemic responses of immediate early genes, and genes involved in modulating apoptosis and remodelling/angiogenesis. In summary, aging is associated with shifts in cardiovascular gene expression consistent with the phenotypic features of older hearts. Reduced tolerance with age may be related to modification of signalling (particularly WNT and TGF-beta), and shifts in expression of immediate early genes, and genes important in control of cell death/survival, angiogenesis, and cardiac remodelling. PMID- 16343837 TI - Rapid and specific identification of four Agrobacterium species and biovars using multiplex PCR. AB - On the basis of 23S rRNA gene sequences, 1 universal forward and 4 taxon (species/biovar)-specific reverse primers were designed for multiplex PCR to aid in identification and differentiation of Agrobacterium rubi, Agrobacterium vitis and Agrobacterium biovars 1 and 2. In reactions with DNA of 119 bacterial strains belonging to: Agrobacterium, Allorhizobium, Mesorhizobium, Rhizobium, Sinorhizobium and Phyllobacterium, as well as phytopathogenic bacteria representing various genera, the primers developed for identification of A. vitis, A. rubi or Agrobacterium biovar 1 amplified only DNA of strains belonging to these taxa, producing fragments of the expected sizes: 478, 1006 and 184bp, respectively. However, in the case of the primer developed for identification of Agrobacterium biovar 2, the characteristic 1066bp PCR product was obtained not only with DNA of this biovar, but also with DNA of 3 atypical biovar 1 strains and some rhizobial strains. Differentiation between Agrobacterium biovar 2 and the other strains was possible using the restriction analysis of this product with endonuclease Alw26I. The method developed is an excellent tool for rapid classification of these 4 taxa of Agrobacterium. PMID- 16343836 TI - Aging-related changes of intracellular Ca2+ stores and contractile response of intestinal smooth muscle. AB - In this study, we investigated the effect of aging on intracellular Ca2+ stores, as sarcoendoplasmic reticulum (SR) and mitochondria, and the influence of these compartments on contraction of rat colon smooth muscle [Bitar, K.N., 2003. Aging and neural control of the GI tract V. Aging and gastrointestinal smooth muscle: from signal transduction to contractile proteins. Am. J. Physiol. Gastrointest. Liver. Physiol. 284(1), G1-G7; Marijic, J., Li, Q.X., Song, M., Nishimaru, K., Stefani, E., Toro, L., 2001. Decreased expression of voltage-and Ca2+-activated K+ channels in coronary smooth muscle during aging. Circ. Res. 88, 210-234; Rubio, C., Moreno, A., Briones, A. Ivorra, M.D., D'Ocon, P., Vila, E., 2002. Alterations by age of calcium handling in rat resistance arteries. J. Cardiovasc. Pharmacol. 40(6), 832-840]. Calcium stores and contraction were evaluated by simultaneous measurements of fluorescence and tension in smooth muscle strips loaded with fura-2. Results showed that activation of muscarinic receptors by methylcholine (MCh, 10 microM), induced a greater contraction in aged rats than in adult animals. The inhibition of Ca2+ ATPase by thapsigargin (TG, 1 microM) did not prevent the refilling of SR either in adult or aged rats. MCh, in the presence of TG, induced an increase in transient fluorescence, indicating a release of Ca2+ from TG-insensitive compartment. The mitochondrial uncoupler, FCCP (5 microM), caused a greater increase in intracellular Ca2+ and tension in aged rats, indicating that mitochondria may accumulate more Ca2+ during aging. The present results show that changes in intracellular Ca2+ stores, such as mitochondria and SR, affect contraction and may cause dysfunctions during aging that could culminate in severe alterations of Ca2+ homeostasis and cell damage. PMID- 16343838 TI - LC-MS characterization of terpene lactones in plasma of experimental animals treated with Ginkgo biloba extracts Correlation with pharmacological activity. AB - Liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry (LC/APCI-ITMS) was applied to determine the concentration of terpene lactone in plasma of guinea pigs after chronic administration of Ginkgo biloba extract enriched in ginkgoterpenes in free form (IDN 5380) or complexed with soy phosphlipids (IDN 5381). Oral treatment of the animals with ginkgoterpenes resulted to inhibit the bronchoconstriction (ITP) and the concomitant increase of the levels of thromboxane B2 (TXB2) in the circulation caused by histamine (HIST) and platelet activating factor (PAF) in normal guinea pigs or by ovalbumin (OA) in actively sensitized guinea pigs. To compare the protective activities of G. biloba forms (IDN 5380 and IDN 5381), ED50 and dose ratio (DR) values for both parameters (ITP and TXB2) were evaluated. The phytosomic form (IDN 5381) significantly reduced (two- to four-fold as compared to free form, P < 0.001) the HIST, PAF or OA-induced airway changes and TXB2 release. In addition it has been observed that the absence of ginkgolide C (GC) in plasma samples (in human and animals) was due to its rapid methylation. PMID- 16343839 TI - A rapid HPLC-ESI-MS/MS for qualitative and quantitative analysis of saponins in "XUESETONG" injection. AB - 'XUESETONG' injection, one of the most widely used proprietary medicines in traditional Chinese medicine, consists of total saponins made from Panax notoginseng, which is a highly valued and important Chinese medicinal herb. It is used to treat cardiovascular diseases. In order to control the quality of XUESETONG injection, a rapid HPLC-ESI-MS/MS method was developed for qualitative and quantitative determination of the saponins. The analyses were performed on SB C18 column using gradient elution in 25 min. Full scan and time programmed selected reaction monitoring (SRM) were used for qualitative and quantitative analysis of saponins, respectively. Twenty-seven saponins were identified and nine of them including notoginsenoside R1, ginsenoside Rb1, Rb2, Rb3, Rc, Rd, Re, Rf and Rg1 were quantified. Ten XUESETONG injections were analyzed and compared. The results showed that there is a great variation among different samples. In conclusion, the developed method is rapid, accurate and sensitive for qualitative and quantitative analysis of saponins in XUESETONG injection. Moreover, it also can be used for the quality control of P. notoginseng raw material and its preparations. PMID- 16343840 TI - Determination of glycosides and sugars in Moutan Cortex by capillary electrophoresis with electrochemical detection. AB - A method based on capillary electrophoresis with electrochemical detection has been developed for the separation and determination of paeoniflorin, sucrose, paeonoside, glucose, and fructose in Moutan Cortex for the first time. Effects of several important factors such as the concentration of NaOH, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter copper disc electrode at a working potential of +0.60 V (versus saturated calomel electrode (SCE)). The five analytes can be well-separated within 12 min in a 40 cm length fused silica capillary at a separation voltage of 12 kV in a 75 mM NaOH aqueous solution. The relation between peak current and analyte concentration was linear over about 3 orders of magnitude with detection limits (S/N = 3) ranging from 0.9 to 1.3 microM for all analytes. The proposed method has been successfully applied to monitor glycoside and sugar contents in the real plant samples with satisfactory assay results. PMID- 16343841 TI - The assessment of cognitive function in advanced HIV-1 infection and AIDS dementia complex using a new computerised cognitive test battery. AB - The validity of a new computerised battery called CogState was determined in 60 individuals with advanced human immunodeficiency virus-1 (HIV-1) infection from which eleven were assessed as AIDS dementia complex (ADC) stage 1 or 2. Twenty one seronegative individuals were recruited as controls. Participants were evaluated with a brief computerised examination, lasting 10-15 min, assessing reaction time, accuracy in working memory and learning. They were also assessed with a standard neuropsychological examination lasting 1h 30 min on average. The computerised assessment demonstrated a good sensitivity of 81.1% and specificity of 69.9% as well as good positive predictive value (81%) and acceptable construct validity (.45-.62). Slowed reaction time and learning deficits in the computerised battery were characteristic of ADC. This study supports the utility of a brief computerised battery in the detection of HIV-associated neurocognitive impairment that could be used for wide-scale screening. PMID- 16343842 TI - An evaluation of social and adaptive skills in adults with bipolar disorder and severe/profound intellectual disability. AB - The purpose of this study was to explore the interrelationship of social and adaptive skills in adults with bipolar disorder and severe or profound intellectual disability. A bipolar group (N=14), a severe psychopathology group without bipolar disorder (N=14), and a control group with no DSM-IV Axis I diagnosis (N=14) were compared on the Vineland Adaptive Behavior Scale (VABS) and the Matson Evaluation of Social Skills for Individuals with sEvere Retardation (MESSIER). Bipolar patients had significantly more negative verbal social skills than the other two groups on the MESSIER, but no group differences on the VABS were noted. The implications of these data are discussed for a better understanding of bipolar disorder in persons with intellectual disability. PMID- 16343843 TI - The association between a genetic polymorphism of coproporphyrinogen oxidase, dental mercury exposure and neurobehavioral response in humans. AB - We previously described a polymorphism in exon 4 of the gene encoding the heme biosynthetic pathway enzyme, coproporphyrinogen oxidase (CPOX4), which significantly modifies the effect of mercury exposure on urinary porphyrin excretion in humans. Here, we examined potential consequences of this polymorphism ("CPOX4") on performance within neurobehavioral domains, symptoms, and mood that are known to be affected by elemental mercury (Hg degrees ) exposure in human subjects. A behavioral test battery was administered on the day of urine and buccal cell collections for 194 male dentists (DDs) and 233 female dental assistants (DAs) occupationally exposed to Hg degrees for an average of 19 and 10 years, respectively. Subjects had no history of health disorders and were employed for a minimum of 5 years in the dental profession. Respective mean urinary mercury (HgU) levels in DDs and DAs were 3.32 (4.87) microg/l and 1.98 (2.29) microg/l. Corresponding indices of chronic occupational Hg degrees exposure, weighted for historical exposure, were 27.1 (20.6) and 15.2 (12.3). The frequencies of the homogygous common (A/A), heterozygous (A/C), and homozygous polymorphic (C/C) genotypes were 75%, 23% and 2% for DDs and 73%, 25%, and 2% for DAs, respectively. DDs and DAs were evaluated separately. Regression analyses controlled for age, premorbid intelligence, alcohol consumption, and education. Statistically significant associations with HgU (p<0.05) were found for nine measures among DDs (BEES Digit SpanForward and Backward, WMS-R Visual ReproductionN Correct, BEES Symbol DigitRate, BEES Finger TappingDom/Non-dom, and Alternate Partialed, Hand SteadinessFactor1, and BEES Tracking), and eight measures among DAs (BEES Digit SpanForward, BEES Symbol DigitRate, BEES Pattern Discrimination Rate, BEES Trailmaking B, BEES Finger TappingDom/Non-dom, and Alternate Partialed, Hand SteadinessFactor1, and Vibration SensitivityHits). CPOX4 status was associated with four measures in DDs (BEES Spatial SpanForward, BEES Pattern MemoryN Correct, BEES Symbol DigitRate, and BEES VigilanceHit) and five measures in DAs (BEES Digit SpanForward, WMS-R Visual ReproductionsN Correct, BEES Symbol DigitRate, BEES Simple and Choice Reaction TimeMove. Both groups experienced an additive effect (no interaction term) for HgU and the CPOX4 polymorphisms on the DigitRate whereas DAs also had additive effects for BEES Digit SpanForward and for Beck's Depression factor 'Worthlessness'. These exploratory findings suggest that the CPOX4 polymorphism may affect susceptibility for specific neurobehavioral functions associated with mercury exposure in human subjects. PMID- 16343845 TI - Boosted ARTMAP: modifications to fuzzy ARTMAP motivated by boosting theory. AB - In this paper, several modifications to the Fuzzy ARTMAP neural network architecture are proposed for conducting classification in complex, possibly noisy, environments. The goal of these modifications is to improve upon the generalization performance of Fuzzy ART-based neural networks, such as Fuzzy ARTMAP, in these situations. One of the major difficulties of employing Fuzzy ARTMAP on such learning problems involves over-fitting of the training data. Structural risk minimization is a machine-learning framework that addresses the issue of over-fitting by providing a backbone for analysis as well as an impetus for the design of better learning algorithms. The theory of structural risk minimization reveals a trade-off between training error and classifier complexity in reducing generalization error, which will be exploited in the learning algorithms proposed in this paper. Boosted ART extends Fuzzy ART by allowing the spatial extent of each cluster formed to be adjusted independently. Boosted ARTMAP generalizes upon Fuzzy ARTMAP by allowing non-zero training error in an effort to reduce the hypothesis complexity and hence improve overall generalization performance. Although Boosted ARTMAP is strictly speaking not a boosting algorithm, the changes it encompasses were motivated by the goals that one strives to achieve when employing boosting. Boosted ARTMAP is an on-line learner, it does not require excessive parameter tuning to operate, and it reduces precisely to Fuzzy ARTMAP for particular parameter values. Another architecture described in this paper is Structural Boosted ARTMAP, which uses both Boosted ART and Boosted ARTMAP to perform structural risk minimization learning. Structural Boosted ARTMAP will allow comparison of the capabilities of off-line versus on-line learning as well as empirical risk minimization versus structural risk minimization using Fuzzy ARTMAP-based neural network architectures. Both empirical and theoretical results are presented to enhance the understanding of these architectures. PMID- 16343844 TI - Behavioral consequences of developmental iron deficiency in infant rhesus monkeys. AB - Human studies have shown that iron deficiency and iron deficiency anemia in infants are associated with behavioral impairment, but the periods of brain development most susceptible to iron deficiency have not been established. In the present study, rhesus monkeys were deprived of iron by dietary iron restriction during prenatal (n=14, 10 microg Fe/g diet) or early postnatal (n=12, 1.5 mg Fe/L formula) brain development and compared to controls (n=12, 100 microg Fe/g diet, 12 mg Fe/L formula) in behavioral evaluations conducted during the first four months of life in the nonhuman primate nursery. Iron deficiency anemia was detected in the pregnant dams in the third trimester and compromised iron status was seen in the prenatally iron-deprived infants at birth, but no iron deficiency was seen in either the prenatally or postnatally iron-deprived infants during the period of behavioral evaluation. Neither prenatal nor postnatal iron deprivation led to significant delays in growth, or gross or fine motor development. Prenatally deprived infants demonstrated a 20% reduced spontaneous activity level, lower inhibitory response to novel environments, and more changes from one behavior to another in weekly observation sessions. Postnatally deprived infants demonstrated poorer performance of an object concept task, and greater emotionality relative to controls. This study indicates that different syndromes of behavioral effects are associated with prenatal and postnatal iron deprivation in rhesus monkey infants and that these effects can occur in the absence of concurrent iron deficiency as reflected in hematological measures. PMID- 16343846 TI - A composite neural network model for perseveration and distractibility in the Wisconsin card sorting test. AB - A composite artificial neural network model is proposed to simulate the performance of the Wisconsin Card Sorting Test. The Wisconsin Card Sorting Test is a test of executive functions where prefrontal deficits are matched to some quantitative measures such as percentage of perseverative errors and number of failures to maintain set. In this work, the proposed model is used to simulate the performances of healthy subjects and patients with prefrontal involvement particularly on these measures. The model is designed in such a way that one of the subsystems, namely, the Hopfield network, serves as the working memory and the other, the Hamming block, as the hypothesis generator. The results show that the proposed relatively simple model is capable of simulating the wide range of the performances of both normal subjects and prefrontal patients on the Wisconsin Card Sorting Test. While lowering the Hamming distance in the Hamming block gave rise to progressively more perseverative responses, changing the threshold vector of the Hopfield network resulted in more set maintenance failures. The former manipulation disrupts the abstraction or mental flexibility and the latter sustained attention or perseverance both of which are the major functions of the prefrontal system. PMID- 16343847 TI - An alternative approach for neural network evolution with a genetic algorithm: crossover by combinatorial optimization. AB - In this work we present a new approach to crossover operator in the genetic evolution of neural networks. The most widely used evolutionary computation paradigm for neural network evolution is evolutionary programming. This paradigm is usually preferred due to the problems caused by the application of crossover to neural network evolution. However, crossover is the most innovative operator within the field of evolutionary computation. One of the most notorious problems with the application of crossover to neural networks is known as the permutation problem. This problem occurs due to the fact that the same network can be represented in a genetic coding by many different codifications. Our approach modifies the standard crossover operator taking into account the special features of the individuals to be mated. We present a new model for mating individuals that considers the structure of the hidden layer and redefines the crossover operator. As each hidden node represents a non-linear projection of the input variables, we approach the crossover as a problem on combinatorial optimization. We can formulate the problem as the extraction of a subset of near-optimal projections to create the hidden layer of the new network. This new approach is compared to a classical crossover in 25 real-world problems with an excellent performance. Moreover, the networks obtained are much smaller than those obtained with classical crossover operator. PMID- 16343848 TI - Comparison of relevance learning vector quantization with other metric adaptive classification methods. AB - The paper deals with the concept of relevance learning in learning vector quantization and classification. Recent machine learning approaches with the ability of metric adaptation but based on different concepts are considered in comparison to variants of relevance learning vector quantization. We compare these methods with respect to their theoretical motivation and we demonstrate the differences of their behavior for several real world data sets. PMID- 16343849 TI - True atherosclerotic aneurysm of the facial artery. AB - True aneurysms of the facial artery are rare. This is the case report of a diabetic, hypertensive patient with a true aneurysm of the facial artery. PMID- 16343850 TI - A comparison of tongue and soft palate squamous cell carcinoma treated by primary surgery in terms of survival and quality of life outcomes. AB - In the surgical management of oral cancer the resection and reconstruction of the mobile tongue and soft palate are most important if function is to be maintained. The present trend towards primary laryngeal surgery for early disease has emphasized the importance of primary surgery if good functional outcomes can be achieved. This study compares the functional and health-related quality of life outcomes for primary surgery and reconstruction of the anterior tongue and soft palate. From a cohort of 566 patients treated from 1992 to 2002, 118 fitted the criteria for anterior tongue and 44 for soft palate resection. University of Washington Quality of Life scores were available in around three quarters of patients. In terms of speech and swallowing a 3/4 or total anterior glossectomy had a worse outcome than a 1/4 or 1/2. In patients having a 3/4 or total resection of the soft palate however, the results showed a similar outcome to those with 1/4 or 1/2 resection. The functional results of 3/4 and total soft palate reconstruction were superior to 3/4 and total anterior tongue resections and were similar to the whole cohort. This finding extends the role of functional surgery in the oropharynx for which primary radiotherapy is often preferred to preserve function. PMID- 16343851 TI - Chondromyxoid fibroma of the zygoma: a case report. AB - Chondromyxoid fibroma is a rare benign tumour of chondral origin. It usually involves the long bones of the lower extremity, whilst involvement of craniofacial skeleton is extremely unusual. The second case of chondromyxoid fibroma of the zygoma described in literature is presented and the surgical resection of the lesion with tumour-free margins as the key factor for avoiding local recurrence of this tumour is emphasised. PMID- 16343852 TI - A modified repositioning system for segmental resection of the mandible. AB - Mandibular reconstruction is required after segmental resection of the mandible. Several techniques have been proposed but have several drawbacks. A modified system (based on Leibinger's titanium-positioning system) that can reposition the residual mandible easily and accurately without interfering with the reconstructive procedure was developed. This system has been used successfully in more than 10 patients, with no complications. PMID- 16343853 TI - Nerve injury caused by mandibular block analgesia. AB - Fifty-four injection injuries in 52 patients were caused by mandibular block analgesia affecting the lingual nerve (n=42) and/or the inferior alveolar nerve (n=12). All patients were examined with a standardized test of neurosensory functions. The perception of the following stimuli was assessed: feather light touch, pinprick, sharp/dull discrimination, warm, cold, point location, brush stroke direction, 2-point discrimination and pain perception. Gustation was tested for recognition of sweet, salt, sour and bitter. Mandibular block analgesia causes lingual nerve injury more frequently than inferior alveolar nerve injury. All grades of loss of neurosensory and gustatory functions were found, and a range of persisting neurogenic malfunctions was reported. Subjective complaints and neurosensory function tests indicate that lingual nerve lesions are more incapacitating than inferior alveolar nerve lesions. Unlike most mechanical injuries after surgery, injection injuries were not followed by a course of spontaneous improvement of neurosensory and/or gustatory function. This may indicate neurotoxicity as a central aetiological factor. Fifty-four percent of the nerve injuries were associated with Articaine 4%, and a substantial increase in the number of injection injuries followed its introduction to the Danish market. PMID- 16343854 TI - Snookered! Facial infection secondary to occult foreign body. AB - A case is presented of a man suffering from a severe purulent temporofacial infection that was secondary to an occult foreign body. The patient had been impaled on a 13 cm section of snooker cue and was entirely unaware of its presence despite the size. The management of this case is presented and discussed. PMID- 16343855 TI - Serum concentration of dehydroepiandrosterone sulphate in female patients with chronic idiopathic urticaria. PMID- 16343856 TI - Pharmacokinetics of ciprofloxacin XR (1000 mg) versus levofloxacin (500 mg) in plasma and urine of male and female healthy volunteers receiving a single oral dose. AB - The new extended-release formulation of ciprofloxacin (ciprofloxacin XR) was designed for once-daily administration in the treatment of urinary tract infection (UTI). The aim of this study was to compare concentrations in plasma, urinary excretion (UE) and pharmacokinetic parameters of ciprofloxacin XR (1000 mg) versus those of levofloxacin (500 mg) in healthy volunteers receiving a single oral dose. In this randomised crossover study, 12 volunteers (6 males, 6 females) received a single oral dose of 1000 mg ciprofloxacin XR or 500 mg levofloxacin to assess the concentrations (by high-pressure liquid chromatography) in plasma up to 32 h and the UE at intervals up to 36 h. The following pharmacokinetic parameters were studied: C(max), t(max), t(1/2), AUC(plasma0-->infinity), AUC(plasma0-->last), Cl(ren), maximal urinary concentration (U(max)), AUC(urine0-->last) and UE. Both fluoroquinolones were well tolerated. The plasma concentrations of levofloxacin were significantly higher than those of ciprofloxacin XR throughout the study period. The urinary concentrations of ciprofloxacin XR were significantly higher than those of levofloxacin in the first collection interval (0-4 h), whereas the concentrations of levofloxacin were significantly higher than those of ciprofloxacin XR in the five last collection intervals (12-36 h). The median proportions of cumulative renal excretion of the administered dose of the parent drug up to 36 h were 43.1% for ciprofloxacin XR (range, 13.7-50.8%; mean +/- standard deviation (S.D.), 40.5 +/- 9.9%) and 79.8% for levofloxacin (range, 74.0-88.2%; mean +/- S.D., 80.4 +/- 5.5%). C(max), AUC(plasma0-->infinity), AUC(plasma0-->last) and UE were statistically significantly higher in the levofloxacin than in the ciprofloxacin XR phase; t(max), Cl(ren) and U(max) were statistically significantly higher in the ciprofloxacin XR phase than in the levofloxacin phase; and AUC(urine0-->last) and t(1/2) were not statistically different. After an oral administration of ciprofloxacin XR 1000 mg and levofloxacin 500 mg, C(max) and AUC(plasma0- >infinity) were significantly higher in the levofloxacin phase. UE of ciprofloxacin XR 1000 mg once daily, however, was equivalent to that of levofloxacin 500 mg, and overall comparable urinary concentrations and AUC(urine) were reached by both drugs. Therefore, it can be assumed that the two doses investigated can be considered equivalent for the treatment of UTI. PMID- 16343857 TI - Trends in antimicrobial susceptibility in UK centres: the MYSTIC Programme (1997 2002). AB - Trends in antimicrobial susceptibilities in three UK centres participating in the MYSTIC Programme were examined from 1997 to 2002. Isolates were tested using standard methodology to determine the susceptibility breakpoints of meropenem and several other antimicrobial agents including imipenem, ceftazidime, piperacillin/tazobactam, ciprofloxacin and gentamicin. Data are grouped in 2-year blocks. The carbapenems were the most active agents tested against the Enterobacteriaceae (99-100% and 98-100% susceptibility to meropenem and imipenem, respectively) and non-fermenters, including Pseudomonas spp. and Acinetobacter spp. With the exception of susceptibility to ciprofloxacin, which decreased among Enterobacteriaceae at the end of the 6-year period, all antibiotics tested retained their levels of activity. The proportion of extended-spectrum beta lactamase (ESBL)- and AmpC-producing Enterobacteriaceae increased during the study (4.8% and 11.3% in 1997-1998; 7.4% and 16.7% in 2001-2002, respectively). Both meropenem and imipenem retained their potency against these ESBL- and AmpC producing isolates (100% for all time periods). All the other antimicrobial agents tested had much lower susceptibility against these resistant isolates and this decreased further over the 6-year period, with the exception of tazobactam, which maintained its low levels. Although all antibiotics tested retained acceptable activity, the carbapenems remained the most active antimicrobial agents against Gram-negative bacteria, including ESBL- and AmpC-producing isolates. PMID- 16343858 TI - Pharmacokinetic-pharmacodynamic aspects of antimicrobial prophylaxis with teicoplanin in patients undergoing major vascular surgery. AB - A prospective, two-arm, open study assessing plasma exposure to teicoplanin with two different prophylactic regimens (Group A (n = 23), 800 mg pre-operatively versus Group B (n = 24), 400 mg pre-operatively plus two doses of 200 mg 24 h apart) was carried out in patients undergoing major vascular surgery. The intent was to define the feasibility and the possible advantages of the single pre operative high dose in ensuring therapeutically effective plasma concentrations (>10 mg/L) of teicoplanin even during long-lasting operations. At the end of the intervention, mean teicoplanin concentrations (+/-S.D.) were 14.05 +/- 5.13 mg/L and 5.39 +/- 2.13 mg/L in Groups A and B, respectively. At 24 h, average teicoplanin levels were 5.10 +/- 1.25 mg/L and 2.08 +/- 0.73 mg/L in Groups A and B, respectively; at 48 h they declined to 2.86 +/- 0.70 mg/L in Group A, whereas they rose to 2.67 +/- 0.82 mg/L after administration of 2.63 +/- 0.51 mg/kg at 24 h in Group B. Single pre-operative high-dose teicoplanin may ensure effective plasma levels even in cases of very long-lasting operations (>8 h) with no need for intraoperative re-dosing and may enable more appropriate prophylactic exposure than that achievable with the same total dose given in three administrations 24 h apart. PMID- 16343859 TI - Clinical consequences of resistant Candida infections in intensive care. AB - Many issues need to be addressed by the Intensive Care Unit physician in the decision-making process regarding antifungal therapy for Candida infection, most importantly pharmacokinetic concerns and/or organ failure limitations. In addition, the extensive use of antifungal agents can select for Candida spp. that exhibit decreased susceptibility to these agents. However, the risk appears low, even in the case of prophylactic or pre-emptive antifungal therapy, but this needs to be confirmed in large-scale studies. PMID- 16343860 TI - Diminished neurocardiac dynamics associated with antipsychotic-induced extrapyramidal syndrome. AB - The relationship between antipsychotic-induced extrapyramidal syndrome (EPS) and the autonomic neurocardiac function was examined in 57 schizophrenic patients treated with atypical antipsychotics. Comprehensive assessments of EPS and heart rate dynamics were performed. There was a significant negative correlation of non hypokinetic parkinsonism, akathisia, and dyskinesia with several linear and novel non-linear heart rate dynamics measures, suggesting reduced neurocardiac dynamics associated with some forms of EPS. Assessment of heart rate dynamics may be useful for the detection of these adverse effects and may serve as a useful non invasive method providing a dynamic window into the alterations of complex neuronal activity. PMID- 16343861 TI - Striatal dopamine D2/D3 receptor availability in male smokers. AB - It has been reported that the activation of the nicotine receptor evokes central endogenous dopamine release. However, whether smoking affects striatal dopamine D(2)/D(3) availability over the long run has not been well established in vivo. Fifteen male smokers and their matching controls were recruited. Striatal dopamine D(2)/D(3) receptor availability was assessed by single photon emission computed tomography with [(123)]IBZM Iodo-benzaimide. There was no significant difference in striatal D(2)/D(3) receptor availability between smokers and controls. PMID- 16343862 TI - Sensitivity considerations in polarization transfer and filtering using dipole dipole couplings: implications for biomineral systems. AB - The robustness and sensitivities of different polarization-transfer methods that exploit heteronuclear dipole-dipole couplings are compared for a series of heterogeneous solid systems, including polycrystalline tetrakis(trimethylsilyl)silane (TKS), adamantane, a physical mixture of doubly (13)C,(15)N-enriched and singly (13)C-enriched polycrystalline glycine, and a powder sample of siliceous marine diatoms, Thalossiosira pseudonana. The methods were analyzed according to their respective frequency-matching spectra or resultant signal intensities. For a series of (13)C{(1)H} cross-polarization experiments, adiabatic passage Hartmann-Hahn cross-polarization (APHH-CP) was shown to have several advantages over other methods, including Hartmann-Hahn cross-polarization (HHCP), variable-amplitude cross-polarization (VACP), and ramped-amplitude cross-polarization (RACP). For X-Y systems, such as (13)C{(15)N}, high and comparable sensitivities were obtained by using APHH-CP with Lee-Goldburg decoupling or by using the transferred-echo double resonance (TEDOR) experiment. The findings were applied to multinuclear (1)H, (13)C, (15)N, and (29)Si CP MAS characterization of a powder diatom sample, a challenging inorganic-organic hybrid solid that places high demands on NMR signal sensitivity. PMID- 16343863 TI - Analysis of photoinitiated polymerization in a membrane mimetic film using infrared spectroscopy and near-IR Raman microscopy. AB - A method has been developed to investigate the extent of polymer cross-linking that results following in situ photopolymerization of an acrylate-functionalized phospholipid assembly adsorbed onto a stabilized, membrane-mimetic film produced from a polyelectrolyte multilayer (PEM) on polytetrafluoroethylene (PTFE) grafts. The acrylate phospholipid monomer was synthesized, prepared as a unilamellar vesicle, and fused onto closed-packed acyl chains that make up the PEM membrane mimetic barrier on the PTFE graft. Both broad band white light and 514.5 nm laser radiation were used as excitation sources for photoinitiation; eosin Y was used as the photoinitiator. The use of 514.5 nm excitation reduced the time for maximum polymerization of the acrylate lipid from 60 min to 240 s. Infrared spectroscopy was successfully used to analyze the extent of photopolymerization in simplified model acrylate lipid systems; however, this method could not be used to analyze acrylate polymerization in heterogeneous, multicomponent PEM membrane-mimetic barriers on PTFE grafts. A near-infrared Raman microscopy method based on the ratio of the integrated areas of the CC and CN vibrations was shown to provide equivalent information to the IR method for analysis of the extent of polymerization efficiency in acrylate lipids. In addition, it proved feasible to extend this near-IR Raman method to the in situ analysis of the extent of polymerization in a stabilized acrylate lipid membrane on a PEM film in a PTFE vascular graft. This work describes a new approach for generating and analyzing the robustness of a membrane-mimetic coating on biomaterial surfaces, and may improve our ability to predict the long-term stability of polymeric membrane mimetic films on implantable medical devices. PMID- 16343865 TI - Improved protein-adsorption-resistant property of PES/SPC blend membrane by adjustment of coagulation bath composition. AB - To improve surface protein-adsorption-resistant property of polyethersulfone (PES) membranes, soybean phosphatidylcholine (SPC) was added to PES casting solution. The blend membranes were prepared by a phase inversion method in a wet process. The surface of PES/SPC blend membranes was characterized by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). XPS data revealed that the phosphorylcholine (PC) groups were concentrated at the surface by changing the composition of coagulation bath. Addition of N,N dimethylformamide (DMF) in coagulation bath could prolong coagulation time and facilitate the migration of SPC from polymer bulk to membrane surface. The PES/SPC blend membranes dramatically reduced BSA and fibrinogen adsorption compared to PES control membrane due to effective immobilization of PC groups at the surface of PES/SPC blend membranes. PMID- 16343864 TI - Does Puccinelia tenuiflora have the ability of salt exudation? AB - The leaves surface of Puccinelia tenuiflora seedling under stress of different concentration of Na(2)CO(3) was observed with scanning electron microscopy and X ray electron probe micro analyzer. All the results indicated that varied salts crystalline distributing in stomatal apparatus on P. tenuiflora leaves surface could be observed by means of frozen-dried sampling. In the case of no stress, these leaves had many kinds of sediments such as Na, K, Ca, Mg, Zn, Cu, Cl, P, S, Si on their surface. When there was stress of Na(2)CO(3), the percentage content of each sediment would have an non-linear relationship to stress. As the concentration of Na(2)CO(3) increased, the percentage contents of Na, Cu, Zn, P, S generally decreased while that of K, Ca, Mg, Si generally increased. The ratio of K/Na also changed the same way as the later. From these results, we concluded, under stress of alkali and salts, P. tenuiflora leaves could exude salts through their stomata or together with wax secretion and these ways might participate many regulation process of P. tenuiflora leaves cells, for example in ion balancing, osmosis regulating and water metabolizing. PMID- 16343866 TI - Critical coagulation concentration for a suspension of cation-absorptive biocolloids. AB - Critical coagulation concentration (CCC) of a biocolloidal suspension is investigated theoretically by taking into account the influences of cationic absorption in particulate membrane phase, variation in dielectric constant, size of charged species and nonuniform distribution of fixed membrane groups. Here, an increase in both valence and effective radius of the original functional group (OFG) via absorption of electrolyte cation(s) is especially considered. The simulated results indicate that stronger membrane electricity yields a larger electrostatic repulsion and a higher potential energy, which generates a higher CCC. A lower CCC can be resulted from a larger (1) cation-functional group complex (CFGC) for a fixed difference between the radius of CFGC and that of OFG, Delta, (2) number of OFG(s) involved in the formation of a CFGC, (3) Delta for a positively charged CFGC, (4) dielectric constant of main membrane phase, in general, (5) membrane thickness for a constant amount of space-average functional groups, and (6) effective radius of anions. CCC decreases with the following parameters: (1) Delta for a negatively charged CFGC, (2) equilibrium constant of the reaction of cationic absorption, (3) nonuniform feature index of fixed groups, and (4) effective radius of cations. PMID- 16343867 TI - [Eosinophilic granuloma revealed by torticollis: a case report]. AB - Langerhans cell histiocytose is a rare condition in childhood. It presents in different ways ranging from a single bony disease to a multisystemic disease involving vital organs. CASE REPORT: We report a case of single bone involvement revealed by torticollis in an eight-year-old boy. The diagnosis was evocated on radiological findings and confirmed by histologic aspects. After a period of 2,5 years, this child is in total spontaneous remission. CONCLUSION: Torticollis must be explored and watched. Eosinophilic granuloma can be a rare aetiology in children. The outcome is often favorable. PMID- 16343868 TI - [Endoscopic therapy for pancreatitis in children]. AB - Authors report on 3 cases of children treated by therapeutic endoscopic retrograde cholangiopancreatography for pancreatitis (ERCP). The first child presented with familial pancreatitis: he was treated by sphincterotomy and lithiasis extraction. The second child presented with pancreatitis secondary to pancreas divisum: she was treated by sphincterotomy and stunt of small caroncula. The third child presented with severe pancreatitis with pseudocyst: after drainage she was treated by sphincterotomy, Wirsung dilatation and lithiasis extraction. ERCP is a beneficial treatment and may be alternative to surgery in complicated pancreatitis. PMID- 16343869 TI - [Prescriptions and children]. AB - Because of a lack in drug evaluation in children, they receive many off license drugs prescription, which means drugs prescribed in an indication, dosage, formulation or age different from the license terms. However, drug evaluation is crucial in paediatrics population because of pharmacokinetics and pharmacodynamics modifications throughout organism development, which make usually wrong any extrapolation to children of data obtained in adults. Drug safety is generally better in infants except in newborns who are at a particularly high drug-related risk because of high frequency of off license drug use, many drugs association and drugs metabolism immaturity. The lack of evaluation in children, the fact that children are the unique target population of some drugs or the maturation phenomena explain some adverse effects more specific in children. The more the children are young, the more they are exposed to medication or drug utilization errors. Physician must take what the SPC, as found in the Vidal dictionary, mentions about children into account while prescribing. Drugs with a paediatric license must be preferred overall, however it should be kept in mind that a paediatric license means only that the drug is active in the indication of the license but it doesn't position it regarding other therapeutic alternatives. Off labeling prescription should be based on a supposed benefit, which had to be justified if a severe side effect occurred. PMID- 16343870 TI - [Pharmacovigilance of vaccines]. AB - Safety of vaccines must be excellent to make vaccine's strategy acceptable, since it usually has a deferred individual benefit but immediate adverse drug reactions (ADRs). Pharmacovigilance of vaccines after their marketing is crucial because, prior to its availability on the market, the size of clinical trials is insufficient to identify rare or deferred adverse effects. The Pharmacovigilance is based on "spontaneous reporting" of ADRs to the Pharmacovigilance Regional Centre (PVRC) which establishes a relationship between each drug taken by the patient and the ADRs occurrence (imputability). This method is crucial to generate alerts, but under-estimates the real frequency of ADRs (1 to 10% of severe ADRs are reported). Thus pharmacoepidemiology studies are necessary to confirm the alerts identified by spontaneous reporting. ADRs can be specific, related to the antigen of an attenuated alive virus vaccine (lymphocyte meningitis after anti-mumps vaccine) or non-specific, related to a component different from the antigen (aluminium hydroxide involved in the "macrophagic myofasciitis", allergic reactions to neomycin, latex, egg or gelatine). Importance of Pharmacovigilance of vaccines is illustrated. Data, especially case control studies, about the relationship between multiple sclerosis and hepatitis B vaccine are summarised. Data about the relationship between Crohn's disease or autism and MMR vaccine are analysed. As vaccines are used in healthy people, their safety must be excellent to be accepted. To monitor them after their marketing is the unique way to detect rare ADRs. This surveillance is made through reporting of ADRs to the PVRC. However, an active and intensive surveillance of ADRs as the one set up from the marketing of Prevenar should be systematic. PMID- 16343871 TI - [Neonatal epilepsy and inborn errors of metabolism]. AB - Metabolic disorders constitute an important cause of neurologic disease, including neonatal epilepsy. Epilepsy rarely dominates the clinical presentation, which is more frequently associated with other neurologic symptoms, such as hypotonia and/or vigilance disturbances. In most cases, epilepsy secondary to inherited metabolic disorders presents with polymorphic clinical and electrographic features that are difficult to classify into precise epileptic syndromes. However, specific types of seizures, such as myoclonic seizures or distinctive electroencephalographic patterns, such as suppression burst patterns, epileptic syndrome or early myoclonic encephalopathy, may suggest a specific metabolic disease. The aim of this article is to help clinicians in reviewing potential metabolic diagnoses and approaching metabolic evaluations. PMID- 16343872 TI - Knowledge-discovery incorporated evolutionary search for microcalcification detection in breast cancer diagnosis. AB - OBJECTIVES: The presence of microcalcifications (MCs), clusters of tiny calcium deposits that appear as small bright spots in a mammogram, has been considered as a very important indicator for breast cancer diagnosis. Much research has been performed for developing computer-aided systems for the accurate identification of MCs, however, the computer-based automatic detection of MCs has been shown difficult because of the complicated nature of surrounding of breast tissue, the variation of MCs in shape, orientation, brightness and size. METHODS AND MATERIALS: This paper presents a new approach for the effective detection of MCs by incorporating a knowledge-discovery mechanism in the genetic algorithm (GA). In the proposed approach, called knowledge-discovery incorporated genetic algorithm (KD-GA), the genetic algorithm is used to search for the bright spots in mammogram and a knowledge-discovery mechanism is integrated to improve the performance of the GA. The function of the knowledge-discovery mechanism includes evaluating the possibility of a bright spot being a true MC, and adaptively adjusting the associated fitness values. The adjustment of fitness is to indirectly guide the GA to extract the true MCs and eliminate the false MCs (FMCs) accordingly. RESULTS AND CONCLUSIONS: The experimental results demonstrate that the incorporation of knowledge-discovery mechanism into the genetic algorithm is able to eliminate the FMCs and produce improved performance comparing with the conventional GA methods. Furthermore, the experimental results show that the proposed KD-GA method provides a promising and generic approach for the development of computer-aided diagnosis for breast cancer. PMID- 16343873 TI - Distributed, intelligent, interactive visualization and exploration of time oriented clinical data and their abstractions. AB - OBJECTIVES: We present KNAVE-II, an intelligent interface to a distributed architecture specific to the tasks of query, knowledge-based interpretation, summarization, visualization, interactive exploration of large numbers of distributed time-oriented clinical data, and dynamic sensitivity analysis of these data. KNAVE-II main contributions to the fields of temporal reasoning and intelligent user interfaces are: (1) the capability for interactive computation and visualization of domain specific temporal abstractions, supported by ALMA--a computational engine that applies the domain knowledge base to the clinical time oriented database. (2) Semantic (ontology-based) navigation and exploration of the data, knowledge, and temporal abstractions, supported by the IDAN mediator, a distributed architecture that enables runtime access to domain-specific knowledge bases that are maintained by expert physicians. METHODS AND MATERIALS: KNAVE-II was designed according to 12 requirements that were defined through iterative cycles of design and user-centered evaluation. The complete architecture has been implemented and evaluated in a cross-over study design that compared the KNAVE-II module versus two existing methods: paper charts and an Excel electronic spreadsheet. A small group of clinicians answered the same queries, using the domain of oncology and a set of 1000 patients followed after bone-marrow transplantation. RESULTS: The results show that users are able to perform medium to hard difficulty level queries faster and more accurately by using KNAVE-II than paper charts and Excel. Moreover, KNAVE-II was ranked first in preference by all users, along all usability dimensions. CONCLUSIONS: Initial evaluation of KNAVE-II and its supporting knowledge based temporal-mediation architecture, by applying it to a large data base of patients monitored several years after bone marrow transplantation (BMT), has produced highly encouraging results. PMID- 16343874 TI - A patient's perspective on medical records. PMID- 16343875 TI - Controlled release of saccharides from matrix tablets. AB - The aim of this study was to design site specific, controlled release tablets of N-acetyl-d-glucosamine (NAG), maltose monohydrate and maltopentaose by using hydrophobic matrix formers starch acetate (SA) and ethyl cellulose (EC). The optimized matrices, which had either low porosity and high drug load or high porosity and low drug load, released the saccharides within the desired 2-4 h. In general, it was possible to control the release rate of saccharides by altering the relative amount of hydrophobic matrix former in the tablet and tablet porosity. The release type of saccharides from these formulations varied from immediate release to sustained release. In the case of sustained release formulations, it was found that the release of maltose monohydrate and maltopentaose was biphasic and slower than the release rate of NAG from similar tablets. NAG release kinetics followed square root of time kinetics, while in the case of maltose monohydrate and maltopentaose, the release kinetics were zero order in both phases. The biphasic dissolution profile was proposed to be caused by water mediated recrystallisation of the disordered material formed during the dissolution. Both SA and EC matrices were found to represent suitable controlled oral delivery vehicles for saccharides. PMID- 16343876 TI - Clinical chemistry and haematology historical data in control Sprague-Dawley rats from pre-clinical toxicity studies. AB - The purpose of this paper is to provide historical data pertaining to clinical chemistry and haematology parameters, obtained from control Sprague-Dawley rats, used in pre-clinical toxicity studies. Mean, standard deviation, minimum and maximum values for haematological and coagulative profiles, haemato-biochemistry and urine analysis data, and the differences per sex and study duration, 4 versus 13 weeks, are presented. The studies were conducted in agreement with the GLP (Good Laboratory Practice) regulations. Statistically significant differences, at the confidence level of 99%, for the red blood cell (RBC) parameters, the white blood cell (WBC) series parameters, plasmatic albumin/globulin (A/G), alanine amino-transferase (ALT), alkaline phosphatase (ALP), creatinine, globulin, glucose, sodium, total protein, tryglycerides, urea and urine volume were observed in males, when 4-week study values were compared with those obtained from 13-week studies. Female rats showed statistically significant variations, at the confidence level of 99% for RBC number and mean corpuscular haemoglobin (MCH), mean red blood cell volume (MCV), WBCs count and lymphocytes percentage, A/G, albumin, ALT, AST, ALP, creatinine, globulin, and sodium, when 4-week study values were compared to 13-week studies. Similar differences were observed comparing the female with male haematological and biochemical data for the two different times of the sample collection. These data could be useful as a reference for evaluation of background pathology in Sprague-Dawley rats, when used in studies performed to evaluate the toxicological profile of a new chemical entity (NCE) in agreement with requirements from international regulatory agencies. PMID- 16343877 TI - Understanding the elasticity of fibronectin fibrils: unfolding strengths of FN III and GFP domains measured by single molecule force spectroscopy. AB - While it is well established that fibronectin (FN) matrix fibrils are elastic, the mechanism of fibril elasticity during extension is still debated. To investigate the molecular origin of FN fibril elasticity, we used single molecule force spectroscopy (SMFS) to determine the unfolding behavior of a recombinant FN III protein construct that contained eight FN-III domains ((1-8)FN-III) and two green fluorescent protein (GFP) domains. FN-III domains were distinguished from GFP domains by their shorter unfolding lengths. The unfolding strengths of both domains were determined for a wide range of pulling rates (50 to 1,745 nm/s). We found that the mechanical stabilities of FN-III and GFP domains were very similar to each other over the entire range of pulling speeds. FN fibrils containing GFP remain brightly fluorescent, even when stretched, meaning that GFP domains remain largely folded. Since GFP and FN-III have equal unfolding strengths, this suggests that FN-III domains are not extensively unraveled in stretched FN fibrils. Our results thus favor an alternative model, which invokes a conformational change from a compact to an extended conformation, as the basis for FN fibril elasticity. PMID- 16343878 TI - Inhibition of prostaglandins does not reduce the cardiovascular changes during endotoxemia in rats. AB - Vasodilatory prostanoids, such as prostacyclin and PGE2, and pro-inflammatory cytokines are known to play a central role in the pathogenesis of endotoxemia. This study was undertaken to elucidate whether indomethacin (INDO), a non selective COX inhibitor, has protective effects against the cardiovascular alterations that occur during endotoxemia. Sprague-Dawley rats were injected intraperitoneally with 15 mg/kg lipopolysaccharide (LPS). LPS injection led to a prominent decrease in cardiac left ventricular end diastolic area (LVEDA) and increased LV fractional shortening (FS), as measured by echocardigraphy. LPS also led to a significant increase in plasma and myocardial TNF-alpha and IL-1beta levels, and elevated plasma and hypothalamic levels of PGE2. Neither the decrease in LVEDA and the increase in FS, nor the elevation in plasma and myocardial cytokine levels were altered by INDO (10 mg/kg). On the other hand, pretreatment with INDO significantly reduced the elevation in PGE2 and the hypothermia induced by LPS. Taken together, this study demonstrates that solely inhibiting the production of PGE2 is not sufficient to reduce the cardiovascular alteration seen in endotoxemia. PMID- 16343879 TI - Relationships between fatty acids and psychophysiological parameters in depressive inpatients under experimentally induced stress. AB - Fatty acids can influence important cellular and hormonal processes in the human body. Non-adequate contents of fatty acids, e.g., in blood, can cause and/or result in various diseases. In depressive patients, changes in fatty acid concentrations were found (deficits in omega3-fatty acids, in particular). This paper poses the question whether there are any relations between psychophysiological parameters and changes in fatty acid compositions. The concentration of fatty acids in serum of 118 psychiatric inpatients measured directly before and after experimentally induced stress of about 1h were analysed in relation to psychophysiological parameters continuously registered during the experimental sessions at admission, discharge and at 3 months follow-up. Systolic and diastolic blood pressure, finger pulse amplitude, forehead temperature (FD) and the EMG activity of the musculus zygomaticus consistently correlated with concentrations of single unsaturated oleic (18:1n-9) and erucic acid (22:1) and saturated myristic (14:0) and lauric acid (12:0). Negative relations were found between FD and the concentration of arachidonic acid (20:4n-6) as well as of palmitoleic acid (16:1). Furthermore, the higher the concentration of the erucic acid at discharge the higher the depression score as assessed by the Beck depression inventory (BDI). High concentrations of palmitoleic acid and lauric acid were related to a low level of depression (BDI and Hamilton scores). The implications of these findings for add-on treatment regimens in depression are discussed. PMID- 16343880 TI - Processing and presentation of tumor antigens and vaccination strategies. AB - Various new aspects of antigen processing have been uncovered through the study of tumor antigens. One of these is the production of antigenic peptides by splicing of two distinct peptide fragments. In one of the two cases reported, the proteasome was found to be responsible for splicing. The presentation of another peptide on major histocompatibility complex class I molecules was found to depend on secretion and reuptake of the parent protein, implying a cross-presentation pathway in melanoma cells. Class II presentation of peptides derived from cytosolic proteins now appears to result from autophagy. Based on increased knowledge on antigen processing and presentation, new vaccination strategies are aimed at improving the targeting of antigens to dendritic cells, promoting cross priming (for example, using chloroquine), improving peptide binding to class I molecules and targeting antigens to both the class I and the class II pathways. PMID- 16343882 TI - Antigen processing and recognition. PMID- 16343883 TI - Pentraxins as a key component of innate immunity. AB - Pentraxins are a complex superfamily of multifunctional molecules characterized by a multimeric structure. C-reactive protein and pentraxin 3 (PTX3) are prototypic molecules of the short and long pentraxin family, respectively. PTX3 is conserved in evolution and produced by innate immune cells. Evidence suggests that PTX3 acts as a non-redundant component of the humoral arm of innate immunity, downstream of, and complementary to, cellular recognition, as well as a tuner of inflammation. PMID- 16343884 TI - Systems biology of neutrophil differentiation and immune response. AB - Systems biology has emerged as a new scientific field, which aims at investigating biological processes at the genomic and proteomic levels. Recent studies have unravelled aspects of neutrophil differentiation and immune responses at the systems level using high-throughput technologies. These studies have identified a plethora of novel effector proteins stored in the granules of neutrophils. In addition, these studies provide evidence that neutrophil differentiation and immune response are governed by a highly coordinated transcriptional programme that regulates cellular fate and function in a context dependent manner. PMID- 16343885 TI - Post-translational modifications of naturally processed MHC-binding epitopes. AB - A variety of different post-translational modifications of peptides displayed by class I and II MHC molecules have now been described. Some modifications promote the binding of peptides to MHC molecules, and might also influence the ability of the peptide to be produced by antigen processing pathways. In some instances, the antigen processing components themselves are actually responsible for generating post-translational modifications. Finally, evidence is accumulating that modifications can be altered as a consequence of inflammation, transformation, apoptosis and aging. This leads to altered repertories of MHC-associated peptides, which may be important in immune responses associated with autoimmune diseases, infection and cancer. PMID- 16343886 TI - How Toll-like receptors signal: what we know and what we don't know. AB - Signal transduction pathways activated by Toll-like receptors (TLRs) have continued to be a major focus of research for investigators interested in the initiation of innate immune responses and the induction of pro-inflammatory cytokines and type I interferons during infection. Biochemical details of the major signalling pathways have now been obtained, and the specific signalling pathways activated by different TLRs are being elucidated. New insights into the activation of IRF family members, notably IRF3, IRF5 and IRF7, have been obtained, and interesting spatiotemporal aspects of signalling by MyD88 leading to IRF7 activation revealed. Adapters in TLR signalling are targets for inhibition, both by endogenous regulators and by virally derived proteins. Selective targeting of pathways by anti-inflammatory glucocorticoids also indicates the potential for modulating TLR signalling therapeutically. PMID- 16343887 TI - Hospital use for COPD patients during the last few years of their life. AB - Little is known about what happens to COPD patients during their final years of life, in particular in terms of hospital use. We linked the Swedish Mortality and Inpatient Registers to examine this research question during the period 1987 2000. In year 2000, 2331 fatalities were diagnosed with COPD as the underlying cause of death. About 3% of individuals had not been admitted to hospital at all prior to their death, whereas 15% had never been admitted for COPD (but for other reasons). More than 68% of all COPD admissions and 74% of all days in hospital occurred in the 3.5 years before death, indicating longer stays closer to death. The last 6 months of life accounted for 22% and 28% of all COPD admissions and days, respectively. Other causes accounted for nearly 50% of all admissions, with a more limited increase during the final years. In conclusion, there is a very variable hospital use among these COPD patients. Use increases almost exponentially as the end approaches. Finally, COPD patients often have a number of concomittant diseases which may affect diagnosis and healthcare resource use. The economic and policy implications of these main findings need to be further examined. PMID- 16343888 TI - Local interactions shape plant cells. AB - Plant cell expansion is usually attributed to the considerable osmotic pressure that develops within and impinges upon the cell boundary. Whereas turgor containment within expandable walls explains global expansion, the scalar nature of turgor does not directly suggest a mechanism for achieving the localized, differential growth that is responsible for the diversity of plant-cell forms. The key to achieving local growth in plant cells appears to lie not in harnessing turgor but in using it to identify weak regions in the cell boundary and thus creating discrete intracellular domains for targeting the growth machinery. Membrane-interacting phospholipases, Rho-like proteins and their interactors, an actin-modulating ARP2/3 complex with its upstream regulators, and actin microtubule interactions play important roles in the intracellular cooperation to shape plant cells. PMID- 16343889 TI - Protein complexes regulating Arp2/3-mediated actin assembly. AB - Key steps in regulating actin dynamics are the de novo nucleation and elongation of actin filaments, which can be catalysed by a limited number of proteins and protein complexes. Among these, Arp2/3 complex and formins are the best studied. Arp2/3-complex activity is controlled through signalling-dependent association with nucleation-promoting factors, such as the WASP/WAVE family proteins. A common theme for these molecules, which is well established for WAVEs but is only just beginning to emerge for WASPs, is that they act as coincident detectors of a variety of signalling pathways through the formation of large multi-molecular complexes. PMID- 16343890 TI - Construction of a test embankment using a sand-tire shred mixture as fill material. AB - Use of tire shreds in construction projects, such as highway embankments, is becoming an accepted way of beneficially recycling scrap tires. However, in the last decade there was a decline in the use of pure tire shreds as fill materials in embankment construction, as they are susceptible to fire hazards due to the development of exothermic reactions. Tire shred-sand mixtures, on the other hand, were found to be effective in inhibiting exothermic reactions. When compared with pure tire shreds, tire shred-sand mixtures are less compressible and have higher shear strength. However, the literature contains limited information on the use of tire shred-soil mixtures as a fill material. The objectives of this paper are to discuss and evaluate the feasibility of using tire shred-sand mixtures as a fill material in embankment construction. A test embankment constructed using a 50/50 mixture, by volume, of tire shreds and sand was instrumented and monitored to: (a) determine total and differential settlements; (b) evaluate the environmental impact of the embankment construction on the groundwater quality due to leaching of fill material; and (c) study the temperature variation inside the embankment. The findings in this research indicate that mixtures of tire shreds and sand are viable materials for embankment construction. PMID- 16343891 TI - Immunodetection of pentamer and modified C-reactive protein using surface plasmon resonance biosensing. AB - In clinical practices, the examination of pentamer C-reactive protein (pCRP) is commonly used as a prognostic indicator of the risk of a patient developing cardiovascular disease (CVD). Structural modification of pCRP produces a modified CRP (mCRP) which exhibits different biological activities in the body. In recent years, mCRP has come to be regarded as a more powerful inducer than pCRP, and hence mCRP measurement has emerged as an important indicator for assessing the risk of developing CVD. The surface plasmon resonance (SPR) biosensing technique can be employed to increase the detection accuracy and real-time response when sensing pCRP or mCRP. In this study, three monoclonal antibodies (Mabs), C8, 8D8, and 9C9, are immobilized on a protein G layer for subsequent CRP detection. The experimental results reveal that the Mab C8 reacts with both pCRP and mCRP, the Mab 8D8 with pCRP, and the Mab 9C9 with mCRP. No false signals caused by non specific binding are observed. When detecting pCRP using Mab C8, the SPR bioassay provides sufficient sensitivity to evaluate whether or not a patient is at risk of developing CVD. SPR biosensing provides a viable and accurate approach for the real-time evaluation of pCRP and mCRP levels, and is therefore of considerable benefit in clinical examinations of CPR. PMID- 16343893 TI - Inherent pacemaker function of duodenal GIST. AB - Gastrointestinal stromal tumours (GIST) are thought to derive from interstitial cells of Cajal (ICCs), which are putative pacemaker cells for gut motility. Isolated cells were obtained by enzymatic treatment of human duodenum GIST tissue having a frequent gain-of-function gene mutation. After cell culturing, c-Kit immunoreactivity was preserved and the cells developed long processes. Whole cell patch clamp recordings revealed voltage-dependent outward currents, without transient inward currents. Intracellular Ca(2+) measurements showed oscillation like spontaneous activity in some GIST cells. RT-PCR revealed expression of ion channels (Kv1.1, Kv1.6 and KCNH2; IP3R1, and IP3R2; TRPC1, 3, 6 and 7; Cx43), which have been suggested to play important roles in pacemaker activity. However, SCN5A, a TTX-resistant Na(+) channel known to be expressed in human ICCs, was below detectable levels. These data suggest that GIST cells appear to preserve some, but not all ionic mechanisms underlying pacemaker activity in ICC. PMID- 16343892 TI - Targeted CML therapy: controlling drug resistance, seeking cure. AB - Targeted cancer therapy with imatinib (Gleevec) has the capability to drive chronic myeloid leukemia (CML) into clinical remission. Some patients, particularly those with advanced disease, develop resistance to imatinib. To counteract this problem, two new BCR-ABL kinase inhibitors for imatinib refractory disease are currently in clinical trials: the imatinib derivative AMN107 and the dual-specificity SRC/ABL inhibitor dasatinib. Using imatinib to reduce leukemic burden also facilitates the detailed investigation into how the persistence of CML disease depends on BCR-ABL signaling, particularly within the leukemic stem cell compartment. Mathematical models of drug resistance and disease relapse, in addition to experimental systems that recapitulate crucial aspects of advanced disease have deepened our understanding of CML biology. Together, these advances are contributing to a high level of disease control, and might ultimately lead to disease eradication. PMID- 16343894 TI - Unilateral retinoblastoma, lack of familial history and older age does not exclude germline RB1 gene mutation. AB - Conclusive identification of RB1 mutations in retinoblastoma is predicted to improve the clinical management of affected children and relatives. However, despite clear clinical benefits, RB1 screening remains difficult, most of the alterations being unique and randomly distributed throughout the entire coding sequence. In this report, we present the results of a constitutional RB1 analysis undertaken in our institution over the last four years. The detection of RB1 gene deletion or mutation was performed by Southern blot and sequence analyses in 73 patients (including three families with 2, 3 and 3 probands, respectively). Complementary constitutional chromosome and fluorescent in situ hybridization (FISH) analyses of RB1 gene were applied in cases where hereditary retinoblastoma was suspected despite negative detection. Altogether, germline abnormalities were found in 11% (4/36 patients) of sporadic unilateral retinoblastoma (median age, 21.5 months) and 86% (32/37 patients) of sporadic bilateral or positive familial history retinoblastoma (median age, 5 months). The spectrum of germline alterations found in 31 distinct families included 12 nonsense mutations (39%); 10 point insertions or deletions with frameshift (32%); 4 mutations and 1 deletion affecting splice sites (16%); 2 missense mutations (6%); and 2 large deletions (6%). A total of 15 mutations have not been previously reported. In this small series, splicing mutations were associated with bilateral disease whilst most of the frameshift mutations were identified in patients with an early age at diagnosis, bilateral disease or hereditary forms of the disease. This study confirms that screening for constitutional RB1 mutation should become an integral part of current management of any patient affected by retinoblastoma irrespective of the tumour laterality and familial background. PMID- 16343895 TI - Bioactivity assay of extracts from Calocedrus macrolepis var. formosana bark. AB - Alcoholic extracts from bark of Calocedrus macrolepis var. formosana Florin (Cupressaceae) were extracted successively using n-hexane, dichloromethane, ethyl acetate, 1-butanol and water, which gave 34.8%, 34.1%, 24.1%, 3.3% and 3.7% soluble fractions, respectively. Antioxidation activity of these fractions by DPPH assay and dissimilar IC50 values of the DPPH showed that ethyl acetate fraction had the best antioxidant activity; its IC50 was 2.6 microg/ml. Analyses of the composition and anti-inflammatory activity of the subfractions from n C6H14 fraction showed that the T3 and H5ppt had the best anti-inflammatory activity in LPS-stimulated murine macrophage J774A. 1 cells, respectively; moreover, their major constituent was sugiol (T3 37.1%, H5ppt 81.1%), which at dosages of 10 microg/ml inhibited proIL-1beta protein production completely. Furthermore, the T1 also exhibited anti-inflammatory activity, and its major constituent was ferruginol (above 85.6%). PMID- 16343896 TI - Accumulation of selenium in mixotrophic culture of Spirulina platensis on glucose. AB - Accumulation of Se in mixotrophic culture of Spirulina platensis was investigated in this study. Results indicated that glucose was better than acetate as an organic carbon source for mixotrophic culture of S. platensis. Supplementation of glucose (2 gL(-1)) significantly enhanced the biomass concentration (2.57 gL(-1)) and the production of phycocyanin (0.279 gL(-1)) and allophycocyanin (0.126 gL( 1)) in S. platensis, which were much higher than those of photoautotrophic culture (1.08 gL(-1), 0.119 gL(-1) and 0.042 gL(-1), respectively). Stepwise addition of Se during the growth phase avoided the inhibitory effect of high Se concentration on the growth of S. platensis. The Se enrichment favored the production of phycocyanin and allophycocyanin in the algal cells. The highest Se yield (1033 microgL(-1)) was obtained at an accumulative Se concentration of 250 mgL(-1), with organic Se percentage, biomass concentration, phycocyanin and allophycocyanin yields of 92.3%, 2.55 gL(-1), 0.295 gL(-1) and 0.153 gL(-1), respectively. These results indicated that the application of mixotrophic culture S. platensis with stepwise addition of Se to the medium could offer an effective and economical way for the production of high Se-enriched algal products. PMID- 16343897 TI - N-Glycosyl-thiophene-2-carboxamides: effects on endothelial cell growth in the presence and absence of bFGF--a significant increase in potency using per-O acetylated sugar analogues. AB - Inhibitors of endothelial cell proliferation are of interest in development of therapies for angiogenesis related disease. N-Glucosyl-thiophene-2-carboxamides have been synthesized and evaluated for their effects on proliferation in bovine aortic endothelial cells. Per-O-acetylated-N-glucosyl-thiophene-2-carboxamides showed improved inhibition of both serum and bFGF stimulated uptake of [(3)H]thymidine, when compared to non-acetylated analogues. PMID- 16343898 TI - Alkyl-linked bis-THTT derivatives as potent in vitro trypanocidal agents. AB - The effect of several alkyl-linked bis tetrahydro-(2H)-1,3,5-thiadiazine-2-thione (bis-THTT) on Leishmania donovani, Trypanosoma brucei rhodesiense, and Plasmodium falciparum is reported. Most of the compounds exhibited a potent activity against the three parasitic strains but the best in vitro activity profiles were found against T. b. rhodesiense with IC(50) values ranging between 0.3 and 4 microM for the most active compounds. PMID- 16343899 TI - CpG DNA/zymosan complex to enhance cytokine secretion owing to the cocktail effect. AB - Zymosan, classified among beta-(1-->3)-d-glucans, is produced from the cell wall of yeast and well known to induce proinflammatory cytokines when ingested by immune cells. We found that zymosan forms a complex with immunostimulatory CpG DNA, where both zymosan and CpG DNA can induce cytokine secretion according to the different mechanisms (i.e., recognized by different receptors). The complex activated macrophages and induced cytokine secretion, more efficiently than separate administration of zymosan or CpG DNA. Microscopic observation showed that this increment of the cytokine secretion can be explained by the fact that zymosan and zymosan/CpG DNA complex are up-taken more than naked CpG DNA. Additionally, existence of two different immunostimulants in the same cells may enhance the immunoresponse. This report presents a new strategy to construct a delivering vehicle for CpG DNA and to enhance its activity with the 'cocktail effect' of the two immunostimulants. PMID- 16343900 TI - Synthesis and in vitro antibiotic activity of 16-membered 9-O-arylalkyloxime macrolides. AB - A series of novel 9-O-arylalkyloxime analogs based on three different 16-membered macrolide scaffolds-5-O-mycaminosyltylonolide (OMT), tilmicosin, and 20-deoxy-20 (3,5-dimethyl-1-piperidin-1-yl)-OMT-was synthesized. In vitro antibiotic activities were assayed against Gram-positive Streptococcus pneumoniae and Staphylococcus aureus and Gram-negative Haemophilus influenzae bacterial strains. Analogs derived from OMT (3-15) showed similar or better antibacterial activities against macrolide-susceptible strains and enhanced activities against macrolide resistant strains compared with erythromycin A, tylosin, or OMT. Similar results were observed for tilmicosin 9-O-arylalkyloxime analogs (18-24). In contrast, most of the 20-deoxy-20-(3,5-dimethyl-1-piperidin-1-yl)-OMT analogs (25-33) showed reduced antibacterial activities compared with OMT. Ribosome-binding studies were performed on compounds 12 (OMT derivative), 20 (tilmicosin derivative), and 29 [20-deoxy-20-(3,5-dimethyl-1-piperidin-1-yl)-OMT derivative]. It was found that these compounds interacted with both domain V and domain II of the Escherichia coli 23S rRNA. PMID- 16343901 TI - Triaminotriazine DNA helicase inhibitors with antibacterial activity. AB - Screening of a chemical library in a DNA helicase assay involving the Pseudomonas aeruginosa DnaB helicase provided a triaminotriazine inhibitor with good antibacterial activity but associated cytotoxicity toward mammalian cells. Synthesis of analogs provided a few inhibitors that retained antibacterial activity and demonstrated a significant reduction in cytotoxicity. The impact of serum and initial investigations toward a mode of action highlight several features of this class of compounds as antibacterials. PMID- 16343902 TI - Effects of alpha-tocopherol and related compounds on reactions involving various organic radicals. AB - Effects of alpha-tocopherol, PMC, and a number of the respective sulfur containing analogues on reactions involving various organic radicals were studied. The test compounds were found to interact with alkyl radicals more effectively than with peroxyl radicals. The presence of a sulfur atom in structures of the respective analogues did not produce significant effects on reactivity. Derivatives of 5-hydroxy-1,3-benzoxathiol-2-one and 6-hydroxy-1,4 benzoxathiin-2(3H)-one displayed a high reactivity toward alpha-hydroxyalkyl radicals. PMID- 16343903 TI - Chemiluminescent acridinium-9-carboxamide boronic acid probes: application to a homogeneous glycated hemoglobin assay. AB - Chemiluminescent acridinium-9-carboxamide probes containing 1, 3, 9, and 27 phenylboronic acids were prepared and their chemiluminescent properties evaluated. The relative chemiluminescent signal from the probes varied from 4 to 0.83 x 10(19)counts/mol across the series, while the apparent affinity of the probes for the diabetes marker glycated hemoglobin increased from 211 to 0.43 microM. The dose-dependent modulation of the chemiluminescent intensity of the probes upon binding was used to demonstrate a homogeneous assay for glycated hemoglobin. PMID- 16343904 TI - Analysis of surgical and diagnostic quality at a specialist breast unit. AB - The quality of a specialist breast unit was analysed prospectively, comparing data from 1998, with data from 2003. The analysis is based on 5451 new referrals and 632 major breast cancer operations. The period was characterized by an increasing number of surgical specialists and improvements in diagnostic resources. There was statistically significant improvement in the following parameters: frequency of excision biopsies fell from 17.3% to 10.4%; preoperative cancer diagnosis without excision biopsy rose from 67.4% to 95.1%; percentage of women receiving breast conserving surgery rose from 24.4% to 44.7%. Cases where less than 10 lymph nodes were removed at axillary dissection fell from 5.8% to 0.5%. Severe postoperative bleeding fell from 4.1% to 0.8%, severe wound infection from 4.7% to 1.1% and severe skin necrosis from 3.7% to 0.2%. It is concluded that centralization of surgical and diagnostic expertise at a high volume institution contributes positively to the quality of care. PMID- 16343905 TI - The Conserved Kinases CDK-1, GSK-3, KIN-19, and MBK-2 Promote OMA-1 Destruction to Regulate the Oocyte-to-Embryo Transition in C. elegans. AB - BACKGROUND: At the onset of embryogenesis, key developmental regulators called determinants are activated asymmetrically to specify the body axes and tissue layers. In C. elegans, this process is regulated in part by a conserved family of CCCH-type zinc finger proteins that specify the fates of early embryonic cells. The asymmetric localization of these and other determinants is regulated in early embryos through motor-dependent physical translocation as well as selective proteolysis. RESULTS: We show here that the CCCH-type zinc finger protein OMA-1 serves as a nexus for signals that regulate the transition from oogenesis to embryogenesis. While OMA-1 promotes oocyte maturation during meiosis, destruction of OMA-1 is needed during the first cell division for the initiation of ZIF-1 dependent proteolysis of cell-fate determinants. Mutations in four conserved protein kinase genes-mbk-2/Dyrk, kin-19/CK1alpha, gsk-3, and cdk-1/CDC2-cause stabilization of OMA-1 protein, and their phenotypes are partially suppressed by an oma-1 loss-of-function mutation. OMA-1 proteolysis also depends on Cyclin B3 and on a ZIF-1-independent CUL-2-based E3 ubiquitin ligase complex, as well as the CUL-2-interacting protein ZYG-11 and the Skp1-related proteins SKR-1 and SKR 2. CONCLUSIONS: Our findings suggest that a CDK1/Cyclin B3-dependent activity links OMA-1 proteolysis to completion of the first cell cycle and support a model in which OMA-1 functions to prevent the premature activation of cell-fate determinants until after they are asymmetrically partitioned during the first mitosis. PMID- 16343906 TI - Mothers' recollections of the Paediatric Intensive Care Unit: associations with psychopathology and views on follow up. AB - The aim of this study was to establish rates of posttraumatic stress symptoms in mothers after a child's admission to a Paediatric Intensive Care Unit (PICU) and their views on the potential value of a follow up appointment with PICU staff. Thirty-four mothers completed the Parental Stressor Scale:PICU, the General Health Questionnaire (GHQ-28) and the Impact of Event Scale, 8 months after discharge. In total 18/34 (53%) scored > or =5 on the GHQ-28 and 6/32 (18%) of the sample scored in the severe range (>35) on the Impact of Event Scale. Distress was associated with retrospective reports of stress experienced during admission (p < 0.001) but not with other demographic or medical variables. Mothers who talked about their feelings at the time of the admission had lower posttraumatic stress scores at 8 months (p = 0.02) and 25/34 (74%) mothers would have appreciated the offer of a follow up appointment. Screening for distress during admission with the Parental Stressor Scale:PICU may identify those mothers in greatest need of psychological support. Mothers' recollections of the Paediatric Intensive Care Unit: Associations with psychopathology and views on follow up. PMID- 16343907 TI - New horizons for (p)ppGpp in bacterial and plant physiology. AB - A hyperphosphorylated guanosine nucleotide, (p)ppGpp, was initially identified as the effector molecule responsible for the stringent response in Escherichia coli. However, a rapidly growing number of reports proves that (p)ppGpp-mediated regulation is conserved in many bacteria and even in plants. It is now clear that (p)ppGpp acts as a global regulator during physiological adaptation of the organism to a plethora of environmental conditions. Adaptation is not only essential for surviving periods of stress and nutrient exhaustion but also for the interaction of bacteria with their eukaryotic host, as observed during pathogenesis and symbiosis, and for bacterial multicellular behaviour. Recently, there have been several new discoveries about the effects of (p)ppGpp levels, balanced by RelA-SpoT homologue proteins, in diverse organisms. PMID- 16343908 TI - Synthesis and biological evaluation of dimeric cinnamaldehydes as potent antitumor agents. AB - It has been reported that 2-hydroxycinnamaldehyde and 2-benzoyl-oxycinnamaldehyde inhibited the activity of farnesyl protein transferase, angiogenesis, cell-cell adhesion, and tumor growth in vivo model. In order to improve its anti-tumor activity, dimeric cinnamaldehydes have been synthesized based on 2 hydroxycinnamaldehyde. The synthesized compounds strongly inhibited the growth of human colon tumor cells with GI50 values of 0.6-10 microM. Especially, 2 piperazine derivative blocked in vivo growth of human colon tumor xenograft in nude mice at 10 mg/kg. It was found that their anti-tumor effects induce apoptosis and cell cycle arrest at G2/M phase by the compounds. It was confirmed by detection of apoptosis markers such as activated caspase-3 and cleaved PARP, and cell cycle analysis. The dimeric compounds also inhibited Cdc25B phosphatase which is essential for preinitiating G2/M transition and S phase progression. PMID- 16343909 TI - A comparison of different analytical methods of determining the solid angle of a circular coaxial source-detector system. AB - In this work, seven different analytical expressions for the solid angle subtended by a circular detector for a coaxial circular source are compared. These expressions -- which have been published by different investigators over the course of the last half century -- are compared for previously published geometries, and for geometries planned in future experimental work. As compared to benchmark Monte Carlo calculations, the expressions published by Segre, Knoll, and Ruby, were the most accurate and exhibited the highest degree of self correlation. PMID- 16343910 TI - Monte Carlo simulation of intrinsic count rate performance of a scintillation camera for diagnostic images. AB - This paper describes Monte Carlo simulation of intrinsic count rate performance of a scintillation gamma camera. MCNP Monte Carlo code was employed to calculate pulse height spectrum and detector efficiency. A custom code written in Fortran language was then developed to simulate, by Monte Carlo method, the distortion in pulse height spectrum due to the pile-up effect for paralyzable and nonparalyzable systems. The results of the simulations, compared with the experimental measurement of count rate performance, showed a good agreement between the two different approaches. PMID- 16343912 TI - Nerve regeneration with the use of a poly(l-lactide-co-glycolic acid)-coated collagen tube filled with collagen gel. AB - AIM: The aim of this study was to develop a novel artificial nerve conduit and to evaluate its efficiency based on the promotion of peripheral nerve regeneration in rabbits. MATERIAL AND METHODS: The nerve conduit was made of a poly (l-lactide co-glycolic acid)-coated collagen tube filled with collagen gel. The conduits were implanted into a 15 mm gap in the peroneal nerves of five rabbits. On the contralateral side, the defects were bridged with collagen-filled vein grafts. RESULTS: Twelve weeks postoperatively nerve regeneration was superior to the vein graft in the PLGA-coated collagen tube, both morphologically and electrophysiologically. CONCLUSION: The results indicate the superiority of the PLGA-coated collagen tube over vein grafts. Furthermore, they show that entubulation repair with this type of tube can support nerve regeneration over a nerve gap distance of at least 15 mm. PMID- 16343913 TI - In-line (18)F-fluorodeoxyglucose positron emission tomography with computed tomography (PET/CT) in patients with carcinoma of the sinus/nasal area and orbit. AB - BACKGROUND: Contrast-enhanced computed tomography (CT) and magnetic resonance imaging (MRI) are the standard imaging techniques to evaluate patients with carcinoma in the sinus/nasal area and orbit. The use of positron emission tomography (PET) with (18)F-fluorodeoxyglucose (FDG) in such patients is as yet less well established. PURPOSE: The aim of this study was to assess the clinical impact of PET co-registered with CT (PET/CT). PATIENTS: Evaluation of 21 consecutive patients. METHODS: A retrospective analysis of the whole body PET/CT studies was done. Images were assessed visually without knowing the results of the other imaging technique. Histology and clinical follow-up served to verify lesions. The clinical impact on therapy was assessed together with the physician in charge. RESULTS: All patients underwent PET/CT and CT or MRI for staging (n=9 scans) and restaging (n=17 scans) without treatment between the examinations. PET/CT changed the treatment protocol in 2 patients at staging and in 7 at re staging. Distant metastases were found in 5 and a secondary tumour in 1 patient. CONCLUSIONS: Whole body PET/CT adds clinically important information to CT or MRI, thus, influencing treatment. PMID- 16343914 TI - Submandibular osseous choristoma. AB - PURPOSE: Osseous choristoma is a rare, benign lesion of the oral cavity and is usually found in the tongue. It presents as a tumour-like mass of normal bony structure with mature cells in an abnormal position. The object of this paper is to report one case of osseous choristoma. PATIENT: A 32-year-old male presented with a lesion in the submandibular region, which was treated by excision. After 28 months of follow-up there was no recurrence. CONCLUSION: Upon reviewing the English literature, no previous case of an osseous choristoma located in the submandibular region has been found. Extended clinical and radiographic follow-up is necessary after surgical excision of an osseous choristoma, despite its benign nature. PMID- 16343916 TI - Superficial lobe parotid gland lipoma. AB - The author presents a case of superficial lobe of the parotid gland lipoma. The lipoma is an asymptomatic, slow growing, freely movable and relatively rare soft mass. Preoperative diagnosis is generally difficult. However certain diagnostic methods such as high resolution computed topographic scanning provides very helpful information regarding this benign parotid gland neoplasia. The treatment of choice is superficial parotidectomy that, if performed correctly excludes the possibility of any second attack. PMID- 16343915 TI - Free flap monitoring using simultaneous non-invasive laser Doppler flowmetry and tissue spectrophotometry. AB - BACKGROUND: Early identification of flap failure is an indispensable prerequisite for flap salvage. Although many methods of free flap monitoring are available, there is still no single reliable non-invasive technique for early recognition of flap failure and for differentiation between arterial occlusion and venous congestion. The aim of this study was to investigate the benefits of the tissue oxygen analysis system O(2)C for monitoring patients undergoing maxillofacial reconstruction with fasciocutaneous radial forearm flaps. MATERIAL AND METHODS: In a prospective clinical study the microcirculatory parameters of blood flow, flow velocity, haemoglobin concentration (AU, Arbitrary Units) and oxygen saturation (%) were assessed by clinical means, by laser Doppler flowmetry and tissue spectrophotometry in 61 patients intraoperatively. Measurements were carried out before flap harvest, in the separated radial forearm flap, immediately after anastomoses and up to 14 days after reconstruction. RESULTS: Following anastomosis, blood flow and flow velocity exceeded the level before flap elevation and reached significant differences by the third postoperative day (p<0.05). Oxygen saturation decreased significantly by the third postoperative day and haemoglobin oxygenation showed stable values after performing anastomosis. Simultaneous, non-invasive laser-Doppler flowmetry and tissue spectrophotometry detected vascular complications in all cases with no false positive or false negative results and prior to clinical assessment. CONCLUSION: For the first time this new device allows reliable prediction of venous congestion by an increase of haemoglobin-concentration, and of arterial occlusion by a decrease in blood flow parameters and oxygen saturation. It can thus differentiate the mechanisms of flap failure before clinical assessment. PMID- 16343917 TI - Sialolipoma of the parotid gland. AB - A case of slow-enlarging mass of the parotid region in a 44-year-old male is presented, which proved to be a lipomatous tumour of the parotid gland. The clinical impression was that of a benign salivary gland tumour. The tumour was situated in the deep lobe of the gland, thus a total parotidectomy was performed, with preservation of the facial nerve. Microscopically the lesion was described as consisting of mature adipose tissue, which, however, encompassed both glandular elements and nerve bundles within it. This tumour has been recently recognized as a separate entity under the term 'sialolipoma'. PMID- 16343918 TI - Parotidectomy: assessment of a surgical technique including facelift incision and SMAS advancement. AB - BACKGROUND: Despite its proven safety and its relevance regarding the cosmetic outcome, the SMAS-lifting technique is not a routine procedure for many surgeons. AIM: To compare the classical (subcutaneous flap and neck incision) with the SMAS lifting techniques for parotidectomies from the patient's perspective. PATIENTS AND METHODS: Both procedures are described, tricks are pointed out. In both procedures the posterior branch of the great auricular nerve was not preserved, hence the two procedures were not evaluated regarding sensitivity of the auricle and preauricular area. Forty consecutive patients were asked to classify their concerns before (1-4 months) and 1 year after surgery (10 classical technique and 30 SMAS-lifting technique). RESULT: Before parotidectomy, patients were concerned in a decreasing order with the facial nerve function, the scar, the soft-tissue defect in the dorsal part of the cheek and Frey's syndrome. Following use of the classical technique, patients were concerned in decreasing order with the soft tissue defect, the scar and Frey's syndrome. Following the SMAS technique, no one was concerned with the scar, Frey's syndrome, or the soft tissue defect although a slight asymmetry could still be noticed. CONCLUSION: The SMAS-lifting technique might possibly appear to offer a new standard procedure for parotidectomy, except for malignant tumours or in obese patients. PMID- 16343919 TI - Ancient schwannoma of the parotid gland: a case report and review of the literature. AB - Schwannomas are encapsulated benign tumours arising from nerve sheath cells, of which ancient schwannoma is one of five variants. Since the first description, only a few ancient schwannomas have been reported in different locations in the head and neck region. In the parotid gland, this tumour is very rare. In this report, a 41-year-old female patient with an ancient schwannoma of the parotid gland is presented and the data of this patient compared with other (comparable) cases described in the literature. PMID- 16343920 TI - Surgical approaches to juvenile nasopharyngeal angiofibroma. AB - INTRODUCTION: Juvenile nasopharyngeal angiofibromas are highly vascular, non encapsulated tumours affecting predominantly young males. These lesions are benign histologically but they may become life-threatening with excessive bleeding or intracranial extension. MATERIAL AND METHODS: The surgical approaches to 22 male patients with nasopharyngeal angiofibromas are reviewed. A modification of midfacial degloving performed without rhinoplasty incisions and lateral osteotomies is described. RESULTS: The patients' ages ranged between 9 and 26 years (mean 14.9). Three stage I tumours, 8 stage II tumours, 6 stage III tumours and 5 stage IV tumours were included into this study. All stage I lesions and one stage II lesion were treated via transnasal endoscopic approach. A modified midfacial degloving approach was used for the removal of seven other stage II lesions, all six stage III lesions, and three stage IV lesions. A combined midfacial degloving-infratemporal fossa Fisch C-transcranial approach was the route chosen for the remaining two stage IV lesions. The complications encountered in the postoperative course include temporary facial palsy in one patient (following a Fisch C infratemporal resection), mild crusting in the nasal cavity in 8 patients, and facial paraesthesia in 6 patients whose tumours were resected via midfacial degloving, and rupture of the subpetrous part of the internal carotid artery in one patient. CONCLUSION: The suggested treatment of juvenile nasopharyngeal angiofibroma consists of an endoscopic transnasal approach for early stage lesions, and a modified midfacial degloving for almost all of the advanced lesions. The latter approach is very useful considering surgical exposure, duration of surgery, cosmetic outcome, and morbidity. It can be combined with an infratemporal approach or craniotomy if necessary. PMID- 16343922 TI - Non-invasive diagnostic of cardiac allograft vasculopathy by 31P magnetic resonance chemical shift imaging. AB - BACKGROUND: Coronary angiography is still the gold standard for the diagnosis of cardiac allograft vasculopathy (CAV) for which alternative non-invasive diagnostic approaches are currently investigated. In this study, we assessed whether 31P magnetic resonance chemical shift imaging can diagnose CAV by studying variations in cardiac high-energy phosphates in a population of adult heart transplant recipients. METHODS AND RESULTS: CAV was defined by coronary angiography as the presence of diffuse coronary irregularities with significant concentric narrowing on epicardial or distal coronary arteries. Eight patients with CAV (group A), and 18 patients without CAV (group B) were included in this study and compared to nine healthy volunteers (group C). Patients and volunteers underwent 31P three-dimensional chemical shift imaging to determine the ratio of phosphocreatine (PCr) and adenosine tri-phosphate (ATP). PCr/ATP was significantly lower in group A (1.51+/-0.50) than in groups B and C (1.98+/-0.53 (p=0.003) and 2.14+/-0.31 (p=0.001)), respectively. Time from transplant, number of episodes of acute rejection, and left ventricular ejection fraction (LVEF) were not significantly different between patient groups. A PCr/ATP value of 1.59 was the optimal cut-off value to predict CAV (specificity and sensitivity of 100% and 72%, respectively). CONCLUSION: Clinically, in vivo 31P chemical shift imaging is a promising, non-invasive method to detect the potential modifications of high-energy phosphates related to CAV and to better screen indications for coronary angiograms. This may be relevant for coronary angiography follow-up and adjustments of immunotherapy regimen. PMID- 16343923 TI - Hospital volume: operative morbidity, mortality and survival in thoracotomy for lung cancer. A Spanish multicenter study of 2994 cases. AB - INTRODUCTION: It has been hypothesized that medical procedures performed in high volume units carry less risk and achieve a better outcome. OBJECTIVE: To determine the relationship between the number of interventions and the operative morbidity, mortality and long-term survival in the surgery of bronchogenic carcinoma (BC). PATIENTS AND METHOD: Prospective, multicenter Spanish study was conducted in 19 departments of thoracic surgery on 2994 patients operated on consecutively with the aim of curing BC. The thoracic surgery departments have been classified into three groups, according to the number of interventions performed per year: I (1-43 cases/year; centers=7; n=565; 18.9%), II (44-54 cases/year; centers=6; n=1044; 34.9%) and III (55 or more cases/year; centers=6; n=1385; 46.3%). RESULTS: When the three groups were compared, the frequency of complete surgery was found to be 84% for group I, 76% for group II and 83% for group III (p=0.001, for comparisons between groups I/II and II/III). The pathological stages were identical in the three groups. The overall morbidity and the mortality in all patients or above the age of 75 or in pneumonectomies were not different among the groups. When considering all the patients with prognostic information (n=2758), no differences were found regarding the 5-year survival among the groups. When only patients in postoperative stage I-II and complete resection were evaluated, excluding operative mortality (n=1128), 5-year survival was 0.58 for group I, 0.57 for group II and 0.50 for group III (p=0.06 between groups II and III; p=0.08 between groups I and III). CONCLUSIONS: No significant differences that do not favor the hypothesis that there is increased surgical risk and worse survival in centers having a lower volume were found in this Spanish multicenter study. PMID- 16343925 TI - Multimodality approach in management of malignant pleural mesothelioma. AB - Malignant pleural mesothelioma (MPM) is a solid, locally aggressive tumor, which has been closely linked to asbestos exposure. The survival rate without treatment ranges from 4 to 12 months. Response to chemotherapy and radiation is poor, and surgery is the most effective therapy. There are currently 3000 new MPM cases per year in the United States, with the peak incidence in the United States and Europe expected to occur in the year 2020. The prognosis depends on the stage of the tumor at the time of diagnosis, its histological type, lymph node status, and resection margins. While the diagnosis is often delayed, earlier intervention may improve life expectancy. Single-modality therapy has not been effective in changing the natural history of MPM. As a result, multimodality regimens involving surgery with radiation, chemotherapy, or immunotherapy have been initiated. Multiple modality approach has demonstrated favorable outcome, particularly in patients with epithelial histology, negative resection margins and presence of no metastases to extrapleural lymph nodes. Cisplatin and mitomycin have demonstrated modest efficacy in management of distant tumor recurrence. Cisplatin and gemcitabine regimen as well as cisplatin/pemetrexed followed by 54 Gy of adjuvant hemithorax radiation have been reported to improve the outcome. PMID- 16343926 TI - Hot flushes in breast cancer patients. AB - OBJECTIVES: A literature search was conducted to gather information concerning the pathophysiologic mechanisms leading to hot flushes, their prevalence and severity in breast cancer patients, their influence on quality of life, and the best therapeutic option. METHODS: Relevant studies in English were selected from Medline. RESULTS AND CONCLUSION: Pathophysiologic mechanisms leading to hot flushes are poorly understood. Estrogen withdrawal is considered to have a central role. Also, serotonin and norepinephrine seem to be involved in hot flush induction. Menopause induced by chemotherapy or ovarian ablation, is accompanied by an abrupt decrease in estrogen level, causing vasomotor symptoms. Hot flushes are also a side effect of tamoxifen and aromatase inhibitors. Quality of life in breast cancer patients may be negatively influenced by hot flushes, and therefore, adequate treatment is important. Currently, of the several non hormonal options, the selective serotonin-reuptake inhibitor (SSRI) venlafaxine is the most effective in breast cancer patients. However, studies on interaction between SSRIs and tamoxifen may influence future recommendations. PMID- 16343927 TI - Differential synthesis of two interleukin-1 receptor antagonist variants and interleukin-8 by peripheral blood neutrophils. AB - With a short lifespan and containing only few ribosomes and endoplasmic reticulum structures, neutrophils are thought to have a limited capacity for protein synthesis. We here show that peripheral blood polymorphonuclear neutrophils (PMN) are able react to stimulants with differential production of two interleukin (IL) 1 receptor antagonist (IL-1ra) isoforms, secreted IL-1ra (sIL-1ra) and the 16kDa intracellular form of IL-1ra (icIL-1ra3), as well as IL-8. Neutrophils of a high purity and with a low degree of preactivation upregulate mRNA and de novo synthesize protein of both IL-1ra variants and IL-8 in response to granulocyte macrophage colony-stimulating factor and lipopolysaccharide. The cytokines are differentially regulated and distributed in two intracellular compartments. In comparison with peripheral blood mononuclear cells (PBMC), PMN produce distinctly more sIL-1ra but significantly less IL-8. This may indicate an anti-inflammatory role, enabling PMN to antagonize proinflammatory signals. It is therefore possible that PMN play an important role in immune regulation by counteracting a dysregulation of the inflammatory process. PMID- 16343928 TI - Involvement of mitogen-activated protein kinase and NF-kappaB activation in Ca2+ induced IL-8 production in human mast cells. AB - Interleukin-8 (IL-8) is a potent proinflammatory chemokine that plays an important role in inflammation by activating and recruiting neutrophils, lymphocytes, and eosinophils. To demonstrate the effect of intracellular Ca(2+) on IL-8 production and related signaling, we stimulated human mast cell line HMC 1 with either calcium ionophore A23187 or thapsigargin. Increase of intracellular Ca(2+) resulted in inducing IL-8 gene expression and protein secretion, and addition of EGTA or BAPTA/AM before Ca(2+) stimulation inhibited the induction of IL-8 production. Intracellular Ca(2+) triggered the activation of mitogen activated protein kinase (MAPK) in HMC-1, especially p42 and p44 isoforms of extracellular signal-regulated kinase (ERK) and p38 MAPK, but not c-Jun N terminal kinase (JNK). Pretreatment of MAPK inhibitors (PD98059 and SB203580) markedly blocked Ca(2+)-induced IL-8 production from cells, and anti-inflammatory drugs, such as dexamethasone and cyclosporin A, partially inhibited the activation of ERK1/2. We determined that increased Ca(2+) activates the nuclear translocation of the transcription factor NF-kappaB. NF-kappaB inhibitors blocked the ability of Ca(2+) to induce IL-8 production, and the activation of NF-kappaB was required for intracellular Ca(2+)-induced up-regulation of IL-8. These results suggest that increased intracellular Ca(2+) stimulated p38 and ERK1/2 MAPK signaling cascades result in NF-kappaB activation and IL-8 production in HMC 1 cells. This study is the first to identify the intracellular signaling pathways involved in the Ca(2+)-mediated up-regulation of IL-8 synthesis and release from HMC-1 cells. PMID- 16343929 TI - Susceptibility to exogenously added interferon-beta protein depends on intracellular interferon-beta mRNA level in human glioma cells. AB - Exogenously added human interferon-beta (HuIFN-beta) protein possesses a remarkable antiproliferative activity in human glioma and melanoma. Endogenous HuIFN-beta protein, which is produced by its gene transfer using cationic liposomes, has much more effective antiproliferative activity against these tumors, even in cells resistant to exogenously added HuIFN-beta protein. As the first step to elucidate the possible difference in antiproliferative mechanisms between exogenous and endogenous HuIFN-beta protein, we here investigated the relationship between the intracellular level of its mRNA and susceptibility to exogenously added HuIFN-beta protein. In this study, we used seven human glioma cell lines (SK-MG-1, SK-MG-4, SK-AO2, U87MG, U251SP, U251MG and T98) and one human melanoma cell line (MMAN). At first, we examined the relationship between spontaneous expression of HuIFN-beta mRNA and susceptibility to exogenously added HuIFN-beta protein (50 IU/ml) in human glioma cells and then confirmed a significant correlation between them. Next, we confirmed that administration of 0 100 IU/ml exogenously added HuIFN-beta protein upregulated the HuIFN-beta mRNA in a dose-dependent manner using the RT-PCR technique and that the HuIFN-beta mRNA was suppressed by siRNA for HuIFN-beta in SK-MG-1 and MMAN cells. Furthermore, we confirmed that the siRNA for HuIFN-beta significantly suppressed the antiproliferative effect of SK-MG-1 cells treated with 10-100 IU/ml HuIFN-beta protein and MMAN cells with 25 and 50 IU/ml HuIFN-beta protein. We found this phenomenon in another human glioma cell line, U87MG cells, as well. This finding would suggest that susceptibility to exogenously added HuIFN-beta protein is related to the amount of intracellular HuIFN-beta mRNA in human glioma and melanoma cells. PMID- 16343930 TI - Mitochondrial localization of CNP2 is regulated by phosphorylation of the N terminal targeting signal by PKC: implications of a mitochondrial function for CNP2 in glial and non-glial cells. AB - Both 2',3'-cyclic nucleotide-3'-phosphodiesterase (CNP) isoforms are abundantly expressed in myelinating cells. CNP2 differs from CNP1 by a 20 amino acid N terminal extension and is also expressed at much lower levels in non-myelinating tissues. The functional role of CNP2, apart from CNP1, and the significance for CNP2 expression in non-myelinating tissues are unknown. Here, we demonstrate that CNP2 is translocated to mitochondria by virtue of a mitochondrial targeting signal at the N-terminus. PKC-mediated phosphorylation of the targeting signal inhibits CNP2 translocation to mitochondria, thus retaining it in the cytoplasm. CNP2 is imported into mitochondria and the targeting signal cleaved, yielding a mature, truncated form similar in size to CNP1. CNP2 is entirely processed in adult liver and embryonic brain, indicating that it is localized specifically to mitochondria in non-myelinating cells. Our results point to a broader biological role for CNP2 in mitochondria that is likely to be different from its specific role in the cytoplasm, along with CNP1, during myelination. PMID- 16343931 TI - Lysed cell models and isolated chromosomes for the study of kinetochore/centromere biochemistry in vitro. AB - The centromere or kinetochore functions in both chromosome movement and in regulation of progression through mitosis. It appears likely that the signaling pathways involved are keenly dependent on solid phase cytoskeletal and karyoskeletal scaffolds that may mediate important physical signals such as tension. Understanding these pathways will be greatly aided by reconstructing the signaling in lysed cell models. Here we present approaches to the in vitro study of signaling pathways in mitotic cells, particularly those involved in protein phosphorylation changes at kinetochores that may control cell cycle progression in M phase. PMID- 16343932 TI - Methods to measure ubiquitin-dependent proteolysis mediated by the anaphase promoting complex. AB - The anaphase-promoting complex (APC) or cyclosome is a multi-subunit ubiquitin ligase that controls progression through mitosis and the G1-phase of the cell cycle. The APC ubiquitinates regulatory proteins such as securin and cyclin B and thereby targets them for destruction by the 26S proteasome. Activation of the APC depends on the activator proteins Cdc20 and Cdh1, which are thought to recruit substrates to the APC. In vitro, APC's RING finger subunit Apc11 alone can also function as a ubiquitin ligase. Here, we review different methods that have been used to measure the ubiquitination activity of the APC in vitro and to analyze APC-mediated degradation reactions either in vitro or in vivo. We describe procedures to isolate the APC from human cells or from Xenopus eggs, to activate purified APC with recombinant Cdc20 or Cdh1 and to measure the ubiquitination activity of the resulting APC(Cdc20) and APC(Cdh1) complexes. We also describe procedures to analyze the ubiquitination activity associated with recombinant Apc11. PMID- 16343933 TI - Detection of modification by ubiquitin-like proteins. AB - Ubiquitin and ubiquitin-like proteins (Ubls) are conjugated to many target proteins either as monomeric units or as polymeric chains. There are at least 12 members of the ubiquitin family in the human genome and their conjugation dramatically alters the properties of the modified protein. The presence of highly active proteases that specifically deconjugate Ubls often means that, in the cell, the steady state level of modified protein is low. Detection of protein species modified by Ubls can therefore represent a significant challenge. Here, we describe methods that have been developed to allow detection of Ubl modified proteins both in vivo and in vitro. PMID- 16343934 TI - Analysis of microtubule polymerization in vitro and during the cell cycle in Xenopus egg extracts. AB - Microtubules are dynamic polymers that participate in multiple cellular processes such as vesicular transport and cell division. Microtubule dynamics alter dramatically during the cell cycle. An excellent system to study microtubule dynamics is Xenopus egg extracts since it is a system that is open to manipulation. The extracts can be cycled between mitosis and interphase allowing the study of microtubules in these phases as well as during cell cycle transitions. Here, we provide simple assays to study microtubules in extracts and in vitro using purified components. Protocols are provided for the purification of frog tubulin, microtubule pelleting from extracts and in vitro, assembly of microtubule structures in extracts, and isolation of microtubule-associated proteins from extract. These methods can be used to analyze the effect of a protein of interest on the microtubule cytoskeleton. PMID- 16343935 TI - Purification of mitotic spindles from cultured human cells. AB - In eukaryotes, both chromosome segregation and the determination of the cell division cleavage plane depend on the mitotic spindle apparatus. Spindle malfunctioning can lead to chromosome mis-segregation and cytokinesis defects and hence result in aneuploidy. Thus, the understanding of the structure and function of mitotic spindles is of interest not only from the perspective of basic science, but has implications also for human health and disease. Until recently, this complex microtubule-based structure was studied mainly by cell biological techniques in mammalian cells, by biochemical assays in Xenopus egg extracts, and by genetic approaches in genetically tractable organisms such as yeast, flies, and nematodes. With the rapid development of mass spectrometry and its increasing application to biological problems, it has become possible to subject highly complex structures, such as the mitotic spindle apparatus, to proteomics approaches. Such studies require the isolation of the mitotic spindle, or its substructures, in sufficient amounts and free of excessive contaminants. A number of methods for the isolation of mitotic spindles from mammalian tissue culture cells have been developed in the past. We have compared these methods and found that protocols based on the stabilization of microtubules by taxol were most efficient and reproducible. Here, we describe the further optimization of a taxol based method, originally developed by Zieve and Solomon [Cell 28 (1982) 233-242], and its application to the isolation of human mitotic spindles at a scale suitable for mass spectrometric analysis [G. Sauer, R. Korner, A. Hanisch, A. Ries, E.A. Nigg, H.H.W. Sillje, Mol. Cell. Proteomics 4 (2005) 35-43]. PMID- 16343937 TI - Monitoring the permeability of the nuclear envelope during the cell cycle. AB - In animal organisms the nuclear envelope (NE) dis-assembles during cell division resulting in complete intermixing of cytoplasmic and nuclear compartments. This leads to the activation of many mitotic enzymes, which were kept away from their substrates or regulators by nuclear or cytoplasmic sequestration in interphase. Nuclear envelope breakdown (NEBD) is thus an essential step of mitotic entry and commits a cell to M-phase. NEBD begins with the partial disassembly of nuclear pore complexes, leading to a limited permeabilization of the NE for molecules up to approximately 40 nm diameter. This is followed by the complete disruption of nuclear pores, which causes local fenestration of the double nuclear membrane and subsequently breakdown of the entire NE structure. Here, we describe the use of different sized inert fluorescent tracer molecules to directly visualize these different steps of NEBD in live cells by fluorescence microscopy. PMID- 16343936 TI - Imaging the division process in living tissue culture cells. AB - We detail some of the pitfalls encountered when following live cultured somatic cells by light microscopy during mitosis. Principle difficulties in this methodology arise from the necessity to compromise between maintaining the health of the cell while achieving the appropriate temporal and spatial resolutions required for the study. Although the quality of the data collected from fixed cells is restricted only by the quality of the imaging system and the optical properties of the specimen, the major limiting factor when viewing live cells is radiation damage induced during illumination. We discuss practical considerations for minimizing this damage, and for maintaining the general health of the cell, while it is being followed by multi-mode or multi-dimensional light microscopy. PMID- 16343939 TI - Expression, purification, and characterization of the 4 zinc finger region of human tumor suppressor WT1. AB - Wilm's Tumor gene 1 (WT1) encodes a zinc finger protein with four distinct splice isoforms. WT1 has a critical role in genesis of various cancer types both at the DNA/RNA and the protein level. The zinc-finger DNA-binding capacity of the protein is located in the C-terminal domain. Two recombinant proteins, 6HIS-ZN wt1 and 6HIS-ZN+wt1, corresponding to two alternative splice variants of the C terminal regions of human WT1 (-KTS) and WT1 (+KTS), respectively, were over expressed with hexa-histidine fusion tags in inclusion bodies in Escherichia coli for crystallization studies. A combination of Ni2+-NTA affinity and size exclusion chromatography was applied for purification of the proteins in denaturing conditions. The effects of various buffers, salts and other additives were scrutinized in a systematic screening to establish the optimal conditions for solubility and refolding of the recombinant WT1 proteins. Circular dichroism analysis revealed the expected betabetaalpha content for the refolded proteins, with a notable degradation of the alpha-helical segment in the DNA-free state. Electrophoretic mobility shift assay with double-stranded DNA containing the double Egr1 consensus site 5'-GCG-TGG-GCG-3' confirmed that 6HIS-ZN-wt1 has higher DNA binding affinity than 6HIS-ZN+wt1. PMID- 16343940 TI - Production of enzymatically active recombinant full-length barley high pI alpha glucosidase of glycoside family 31 by high cell-density fermentation of Pichia pastoris and affinity purification. AB - Recombinant barley high pI alpha-glucosidase was produced by high cell-density fermentation of Pichia pastoris expressing the cloned full-length gene. The gene was amplified from a genomic clone and exons (coding regions) were assembled by overlap PCR. The resulting cDNA was expressed under control of the alcohol oxidase 1 promoter using methanol induction of P. pastoris fermentation in a Biostat B 5 L reactor. Forty-two milligrams alpha-glucosidase was purified from 3.5 L culture in four steps applying an N-terminal hexa-histidine tag. The apparent molecular mass of the recombinant alpha-glucosidase was 100 kDa compared to 92 kDa of the native barley enzyme. The secreted recombinant enzyme was highly stabile during the 5-day fermentation and had significantly superior specific activity of the enzyme purified previously from barley malt. The kinetic parameters Km, Vmax, and kcat were determined to 1.7 mM, 139 nM x s(-1), and 85 s(-1) using maltose as substrate. This work presents the first production of fully active recombinant alpha-glucosidase of glycoside hydrolase family 31 from higher plants. PMID- 16343941 TI - Expression, purification, and characterization of a novel recombinant fusion protein, rhTPO/SCF, in Escherichia coli. AB - Thrombopoietin (TPO) is the principal regulatory cytokine of megakaryopoiesis and thrombopoiesis and promotes all aspects of megakaryocyte development. Stem cell factor (SCF) is mainly a pleiotropic cytokine acting on hematopoiesis by promoting the survival and proliferation of hematopoietic stem cells and has a potent synergistic effect on megakaryopoiesis in the presence of TPO. Here, we report the construction, expression, and purification of a novel recombinant human thrombopoietin/stem cell factor (rhTPO/SCF) fusion protein, which consists of a truncated human thrombopoietin (1-157 a.a.) plus a truncated human stem cell factor (1-145 a.a.), linked by a peptide (GGGGSPGGSGGGGSGG). The TPO/SCF gene was cloned into the Escherichia coli expression vector pET28a and expressed in BL21(DE3) strain. The rhTPO/SCF constituted up to 6% of the total bacterial protein. Co-expression with E. coli chaperones, Trigger Factor (TF) and GroES/GroEL, and lowering cultivation temperature cooperatively improved the solubility of expressed rhTPO/SCF, resulting in about fourfold increase in the yield soluble rhTPO/SCF. The rhTPO/SCF was purified to homogeneity using anion exchange followed by metal affinity chromatography. Western blot analysis confirmed the identity of the purified protein. rhTPO/SCF stimulated a dose dependent cell proliferation in both TF1 and Mo7e cell lines. PMID- 16343942 TI - Folate levels and cancer morbidity and mortality: prospective cohort study from Busselton, Western Australia. AB - PURPOSE: To test a community population for the hypothesis that carcinogenesis is related to blood folate levels. METHODS: Prospective analysis of cancer mortality data for a cohort of 964 men (person-time follow up: 20,254 years) and 1024 women (person-time follow up: 24,970 years) and morbidity data for a subcohort, all of whom participated in the 1969 Busselton (Western Australia) Health survey. Outcome measures were adjusted hazard ratios according to baseline folate levels for total cancer mortality and morbidity and site specific mortality and morbidity for colorectal, lung, breast, and prostate cancers. RESULTS: In total, there were 278 cancer deaths--45 from colorectal cancer, 44 from lung cancer, 15 from breast cancer, and 31 from prostate cancer. Decreased serum folate levels showed an independent association with increased prostate cancer mortality risk, the adjusted hazard ratio per decrease of 2 microg/L was 1.56 (CI: 1.05, 2.38), men whose levels were in the lowest quartile had an adjusted hazard ratio of 4.79 (CI: 1.56, 14.43) for subsequent death from prostate cancer. The morbidity subcohort data showed that decreased red blood-cell folate was significantly associated with increased events due to breast cancer, the adjusted hazard ratio per decrease of 100 mug/L was 1.96 (CI: 1.22, 3.12), women in the lowest quartile of red cell folate levels had an adjusted hazard ratio of 6.46 (CI: 1.19, 35.07) for a subsequent breast cancer event. Mortality and morbidity from colorectal or lung cancers were not associated with folate levels. CONCLUSIONS: Independent associations, assessed over periods greater than 20 years, were demonstrated between decreased folate levels and increased risks of prostate cancer mortality and breast cancer morbidity. PMID- 16343943 TI - Towards high-resolution three-dimensional imaging of native mammalian tissue: electron tomography of frozen-hydrated rat liver sections. AB - Cryo-electron tomography of frozen-hydrated specimens holds considerable promise for high-resolution three-dimensional imaging of organelles and macromolecular complexes in their native cellular environment. While the technique has been successfully used with small, plunge-frozen cells and organelles, application to bulk mammalian tissue has proven to be difficult. We report progress with cryo electron tomography of frozen-hydrated sections of rat liver prepared by high pressure freezing and cryo-ultramicrotomy. Improvements include identification of suitable grids for mounting sections for tomography, reduction of surface artifacts on the sections, improved image quality by the use of energy filtering, and more rapid tissue excision using a biopsy needle. Tomographic reconstructions of frozen-hydrated liver sections reveal the native structure of such cellular components as mitochondria, endoplasmic reticulum, and ribosomes, without the selective attenuation or enhancement of ultrastructural details associated with the osmication and post-staining used with freeze-substitution. PMID- 16343945 TI - A novel dual-axis iterative algorithm for electron tomography. AB - A new algorithm for computing electron microscopy tomograms which combines iterative methods with dual-axis geometry is presented. Initial modelling using test data shows several improvements over both the weighted back-projection and simultaneous iterative reconstruction technique methods, with increased stability and tomogram fidelity under high-noise conditions. Preliminary experimental dual axis reconstructions confirm the viability of the new algorithm. PMID- 16343944 TI - Conformational variability of the GTPase domain of the signal recognition particle receptor FtsY. AB - The prokaryotic signal recognition particle Ffh and its receptor FtsY allow targeting of proteins into or across the plasma membrane. The targeting process is GTP dependent and the two proteins constitute a distinct GTPase family. The receptor FtsY is composed of A and NG domains where the NG's GTPase domain plays a critical role in the targeting process. In this study, we describe two X-ray structures determined independently of each other of the NG domain of FtsY from Mycoplasma mycoides (MmFtsY). The two structures are markedly different in three of the nucleotide-binding segments, GI (P-loop), GII, and GIII, making only one of the structures compatible with nucleotide binding. Interestingly, the two distinct conformations of the nucleotide-binding segments of MmFtsY are similar to the apo- and ADP-loaded forms of certain ATPases. The structure of the extended interface between the A and NG domains of MmFtsY provides new insights into the role of the A domain for phospholipid interaction. PMID- 16343946 TI - Single particle reconstruction of the human apo-transferrin-transferrin receptor complex. AB - Most organisms depend on iron as a co-factor for proteins catalyzing redox reactions. Iron is, however, a difficult element for cells to deal with, as it is insoluble in its ferric (Fe3+) form and potentially toxic in its ferrous (Fe2+) form. Thus, in vertebrates iron is transported through the circulation bound to transferrin (Tf) and delivered to cells through an endocytotic cycle involving the transferrin receptor (TfR). We have previously presented a model for the Tf TfR complex in its iron-bearing form, the diferric transferrin (dTf)-TfR complex [Cheng, Y., Zak, O., Aisen, P., Harrison, S.C., Walz, T., 2004. Structure of the human transferrin receptor-transferrin complex. Cell 116, 565-576]. We have now calculated a single particle reconstruction for the complex in its iron-free form, the apo-transferrin (apoTf)-TfR complex. The same density map was obtained by aligning raw particle images or class averages of the vitrified apoTf-TfR complex to reference models derived from the structures of the dTf-TfR or apoTf TfR complex. We were unable to improve the resolution of the apoTf-TfR density map beyond 16A, most likely because of significant structural variability of Tf in its iron-free state. The density map does, however, support the model for the apoTf-TfR we previously proposed based on the dTf-TfR complex structure, and it suggests that receptor-bound apoTf prefers to adopt an open conformation. PMID- 16343947 TI - Having a body versus moving your body: How agency structures body-ownership. AB - We investigated how motor agency in the voluntary control of body movement influences body awareness. In the Rubber Hand Illusion (RHI), synchronous tactile stimulation of a rubber hand and the participant's hand leads to a feeling of the rubber hand being incorporated in the participant's own body. One quantifiable behavioural correlate of the illusion is an induced shift in the perceived location of the participant's hand towards the rubber hand. Previous studies showed that the induced changes in body awareness are local and fragmented: the proprioceptive drift is largely restricted to the stimulated finger. In the present study, we investigated whether active and passive movements, rather than tactile stimulation, would lead to similarly fragmented body awareness. Participants watched a projected image of their hand under three conditions: active finger movement, passive finger movement, and tactile stimulation. Visual feedback was either synchronous or asynchronous with respect to stimulation of the hand. A significant overall RHI, defined as greater drifts following synchronous than asynchronous stimulation, was found in all cases. However, the distribution of the RHI across stimulated and non-stimulated fingers depended on the kind of stimulation. Localised proprioceptive drifts, specific to the stimulated finger, were found for tactile and passive stimulation. Conversely, during active movement of a single digit, the proprioceptive drifts were not localised to that digit, but were spread across the whole hand. Whereas a purely proprioceptive sense of body-ownership is local and fragmented, the motor sense of agency integrates distinct body-parts into a coherent, unified awareness of the body. PMID- 16343948 TI - Controlling for individual differences in fMRI brain activation to tones, syllables, and words. AB - Previous neuroimaging studies have consistently reported bilateral activation to speech stimuli in the superior temporal gyrus (STG) and have identified an anteroventral stream of speech processing along the superior temporal sulcus (STS). However, little attention has been devoted to the possible confound of individual differences in hemispheric dominance for speech. The present study was designed to test for speech-selective activation while controlling for inter individual variance in auditory laterality, by using only subjects with at least 10% right ear advantage (REA) on the dichotic listening test. Eighteen right handed, healthy male volunteers (median age 26) participated in the study. The stimuli were words, syllables, and sine wave tones (220-2600 Hz), presented in a block design. Comparing words > tones and syllables > tones yielded activation in the left posterior MTG and the lateral STG (upper bank of STS). In the right temporal lobe, the activation was located in the MTG/STS (lower bank). Comparing left and right temporal lobe cluster sizes from the words > tones and syllables > tones contrasts on single-subject level demonstrated a statistically significant left lateralization for speech sound processing in the STS/MTG area. The asymmetry analyses suggest that dichotic listening may be a suitable method for selecting a homogenous group of subjects with respect to left hemisphere language dominance. PMID- 16343949 TI - Modelling cardiac signal as a confound in EEG-fMRI and its application in focal epilepsy studies. AB - Cardiac noise has been shown to reduce the sensitivity of functional Magnetic Resonance Imaging (fMRI) to an experimental effect due to its confounding presence in the blood oxygenation level-dependent (BOLD) signal. Its effect is most severe in particular regions of the brain and a method is yet to take it into account in routine fMRI analysis. This paper reports the development of a general and robust technique to improve the reliability of EEG-fMRI studies to BOLD signal correlated with interictal epileptiform discharges (IEDs). In these studies, ECG is routinely recorded, enabling cardiac effects to be modelled, as effects of no interest. Our model is based on an over-complete basis set covering a linear relationship between cardiac-related MR signal and the phase of the cardiac cycle or time after pulse (TAP). This method showed that, on average, 24.6 +/- 10.9% of grey matter voxels contained significant cardiac effects and 22.3 +/- 24.1% of those voxels exhibiting significantly IED-correlated BOLD signal also contained significant cardiac effects. We quantified the improvement of the TAP model over the original model, without cardiac effects, by evaluating changes in efficiency, with respect to estimating the contrast of the effects of interest. Over voxels containing significant, cardiac-related signal, efficiency was improved by 18.5 +/- 4.8%. Over the remaining voxels, no improvement was demonstrated. This suggests that, while improving sensitivity in particular regions of the brain, there is no risk that the TAP model will reduce sensitivity elsewhere. PMID- 16343950 TI - Semantic relevance explains category effects in medial fusiform gyri. AB - We used functional Magnetic Resonance Imaging to explore the neural correlates of semantic relevance in 12 healthy participants performing a picture-naming task. In addition, we tested the hypothesis that category effects typically found in functional imaging can be partly explained in terms of different semantic relevance for animals and artefacts. We report that semantic relevance modulates neuronal responses in the medial fusiform gyrus bilaterally. As predicted, category effects in this region are strongly modulated by the semantic relevance of the items. Specifically, the effect of artefacts > animals is greatly reduced when the two categories are matched for semantic relevance. Thus, the present study demonstrates that neuronal responses during concept retrieval are modulated by the semantic relevance of the features. It also suggests that increased activation in the medial fusiform gyrus typically found for artefacts > animals can be explained by different semantic relevance for animal and artefact items. PMID- 16343951 TI - Spatial resolution, signal-to-noise ratio, and smoothing in multi-subject functional MRI studies. AB - Functional MRI is aimed at localizing cortical activity to understand the role of specific cortical regions, providing insight into the neurophysiological underpinnings of brain function. Scientists developing fMRI methodology seek to improve detection of subtle activations and to spatially localize these activations more precisely. Except for applications in the clinical environment, such as functional mapping in patients prior to neurosurgical intervention, most basic neuroscience studies involve group level random-effects analyses. Prior to grouping data, the data from each individual are typically smoothed. A wide range of motivations for smoothing have been given including to match the spatial scale of hemodynamic responses, to normalize the error distribution (by the Central Limit Theorem) to improve the validity of inferences based on parametric tests, and, in the context of inter-subject averaging smoothing has been shown necessary to project the data down to a scale where homologies in functional anatomy are expressed across subjects. This work demonstrates that, for single-subject studies, if smoothing is to be employed, the data should be acquired at lower resolutions to maximize SNR. The benefits of a low-resolution acquisition are limited by partial volume effects and by the weak impact of resolution-dependent noise on the overall group level statistics. Given that inter-subject noise dominates across a range of tasks, improvements in within-subject noise, through changes in acquisition strategy or even moving to higher field strength, may do little to improve group statistics. Such improvements however may greatly impact single-subject studies such as those used in neurosurgical planning. PMID- 16343952 TI - Catheter confocal fluorescence imaging and functional magnetic resonance imaging of local and systems level recovery in the regenerating rodent sciatic nerve. AB - The goal of the present work was to develop minimally invasive imaging techniques to monitor local regeneration of peripheral nerves and to determine the extent of return to function of brain cortical regions associated with that nerve. The sciatic nerve crush model was applied to Sprague-Dawley rats and conventional histological staining for myelin, axons and cell architecture was carried out, as well as traditional behavioral testing, to verify that nerve regeneration was occurring. The rate of sciatic nerve regeneration was measured by determining the distance a lipophilic, fluorescence probe (DiO) would move along the nerve's membrane following a direct injection into the sciatic nerve. This movement was monitored using a catheter based, confocal fluorescence microscope. Two to five days after the crush, the dye moved 1.4 + 0.6 mm/day, as compared to a distance of 5.3 + 0.5 mm/day in the normal nerve. Between 9 and 13 days following the crush, the distance the dye moved increases to 5.5 + 0.5 mm/day, similar to the control, and by 15 days following the crush, the distance increased to 6.5 + 0.9 mm/day. Functional Magnetic Resonance Imaging (fMRI) measurements were performed on alpha-chloralose anesthetized rats to monitor the return of somatosensory cortical functions, which were activated by the stimulation of the lesioned peripheral nerve. fMRI results showed the return of cortical activation around 15 days following the crush procedure. However, the somatosensory cortical region activated by stimulating the crushed hindpaw was significantly smaller in extent than the intact hindpaw stimulation. These findings demonstrate that fluorescence imaging and fMRI can integrate local and system level correlates of nerve regeneration in a non-destructive manner, thus enabling serial imaging of individual animals. PMID- 16343953 TI - Phylogeny of the Callandrena subgenus of Andrena (Hymenoptera: Andrenidae) based on mitochondrial and nuclear DNA data: polyphyly and convergent evolution. AB - We propose a phylogenetic hypothesis of relationships within Callandrena, a North American subgenus of the bee genus Andrena, based on both mitochondrial and nuclear DNA sequences. Our data included 695 aligned base pairs comprising parts of the mitochondrial genes cytochrome oxidase subunits I and II and the intervening tRNA-leucine and 767 aligned base pairs of the F2 copy of the nuclear gene elongation factor-1alpha. We also suggest a preliminary hypothesis of relationships of the North American subgenera in the genus. Our analyses included 54 species of Callandrena, 42 species of Andrena representing 24 additional subgenera, and 11 outgroup species in the family Andrenidae. Parsimony analyses of each marker separately suggested that Callandrena was polyphyletic, with a combined analysis suggesting that there were at least two phylogenetically independent clades of bees with similar morphological features. Maximum likelihood and Bayesian analyses supported this conclusion, as did the non parametric bootstrapping SOWH test. Convergence in morphological characters was likely due to their common use of members of Asteraceae as pollen hosts. PMID- 16343954 TI - Current methods used to investigate G protein coupled receptor oligomerisation. AB - Classical models of G protein coupled receptor (GPCR) signalling assume that each receptor functions as a single unit. However, evidence is increasing that GPCRs may form functional assemblies of dimeric or oligomeric units. There are several methods that can be used to give evidence of GPCR oligomerisation that will be discussed in this review. These include co-immunoprecipitation and Western blotting, resonance energy transfer methods and transactivation / complementation of partially functional receptors. One definitive method currently does not exist and there are various advantages and disadvantages to each method depending upon the system considered. Although co-immunoprecipitation and Western blot studies require disruption of the cellular environment and require specific antibodies, they are a good starting point to show that receptor oligomerisation occurs in native systems. Resonance energy transfer techniques provide evidence that receptors are in close proximity, are measured in living cells and some formats may be used for imaging applications. Transactivation / complementation requires extensive modification of the GPCR, but provides evidence that the receptors are in physical contact. Despite great advances being made using these techniques, future challenges involve the development of other methodologies to determine the role of receptor complexes in the pharmacology and physiology of native systems. PMID- 16343955 TI - Cognitive functions evaluated by P300 and visual and auditory number assays in children with childhood epilepsy with occipital paroxysms (CEOP). AB - PURPOSE: This study was planned to evaluate cognitive functions, especially attention and immediate recall, in children with childhood epilepsy with occipital paroxysms (CEOP), by using P300 and neuropsychological tests, which included visual and auditory number assays. Thirty patients with CEOP, ages ranging from 5 to 17 years were enrolled in the study. Twenty-five healthy children were taken as the control group. METHODS: Oddball paradigm was used in P300 recordings. The latency and the amplitude of the P300 wave recorded from Cz were taken into consideration. The neuropsychological test battery included visual and auditory number assays. RESULTS: P300 latency was significantly longer in the CEOP group (p=0.014). The results of the visual and auditory number assay test showed significant decline in the patient group when compared with the normal controls. CONCLUSIONS: Attention and immediate recall deficits as well as prolonged P300 latencies in children with CEOP can be due to an ongoing epileptic activity either influencing the whole brain or only the occipital lobe which can also be involved in the neuropsychological organization of the human cortex. Therefore, children with CEOP should be evaluated with more detailed neuropsychological tests for possible cognitive deficits. PMID- 16343956 TI - Application of the MAS-J-HMQC experiment to a new pair of nuclei {29Si,31P}: Si5O(PO4)6 and SiP2O7 polymorphs. AB - We report the results of the two-dimensional MAS-J-HMQC experiment providing scalar correlations between 29Si and 31P nuclei in solid state NMR, and we give the first evaluation of the 2JSi-O-P coupling constants (approximately 15 Hz) for a crystalline silicophosphate phase Si5O(PO4)6. The experiment is applied to the characterization of complex mixtures of SiP2O7 phases, through editing of 31P spin pairs by the heteronuclear 2JP-O-Si interaction. PMID- 16343957 TI - Application of Maximum Entropy reconstruction to PISEMA spectra. AB - Maximum Entropy reconstruction is applied to two-dimensional PISEMA spectra of stationary samples of peptide crystals and proteins in magnetically aligned virus particles and membrane bilayers. Improvements in signal-to-noise ratios were observed with minimal distortion of the spectra when Maximum Entropy reconstruction was applied to non-linearly sampled data in the indirect dimension. Maximum Entropy reconstruction was also applied in the direct dimension by selecting sub-sets of data from the free induction decays. Because the noise is uncorrelated in the spectra obtained by Maximum Entropy reconstruction of data with different non-linear sampling schedules, it is possible to improve the signal-to-noise ratios by co-addition of multiple spectra derived from one experimental data set. The combined application of Maximum Entropy to data in the indirect and direct dimensions has the potential to lead to substantial reductions in the total amount of experimental time required for acquisition of data in multidimensional NMR experiments. PMID- 16343958 TI - Electron spin relaxation of copper(II) complexes in glassy solution between 10 and 120 K. AB - The temperature dependence, between 10 and 120 K, of electron spin-lattice relaxation at X-band was analyzed for a series of eight pyrrolate-imine complexes and for ten other copper(II) complexes with varying ligands and geometry including copper-containing prion octarepeat domain and S100 type proteins. The geometry of the CuN4 coordination sphere for pyrrolate-imine complexes with R=H, methyl, n-butyl, diphenylmethyl, benzyl, 2-adamantyl, 1-adamantyl, and tert-butyl has been shown to range from planar to pseudo-tetrahedral. The fit to the recovery curves was better for a distribution of values of T1 than for a single time constant. Distributions of relaxation times may be characteristic of Cu(II) in glassy solution. Long-pulse saturation recovery and inversion recovery measurements were performed. The temperature dependence of spin-lattice relaxation rates was analyzed in terms of contributions from the direct process, the Raman process, and local modes. It was necessary to include more than one process to fit the experimental data. There was a small contribution from the direct process at low temperature. The Raman process was the dominant contribution to relaxation between about 20 and 60 K. Debye temperatures were between 80 and 120 K. For samples with similar Debye temperatures the coefficient of the Raman process tended to increase as gz increased, as expected if modulation of spin-orbit coupling is a major factor in relaxation rates. Above about 60 K local modes with energies in the range of 260-360 K (180-250 cm-1) dominated the relaxation. For molecules with similar geometry, relaxation rates were faster for more flexible molecules than for more rigid ones. Relaxation rates for the copper protein samples were similar to rates for small molecules with comparable coordination spheres. At each temperature studied the range of relaxation rates was less than an order of magnitude. The spread was smaller between 20 and 60 K where the Raman process dominates, than at higher temperatures where local modes dominate the relaxation. Spin echo dephasing time constants, Tm, were calculated from two-pulse spin echo decays. Near 10 K Tm was dominated by proton spins in the surroundings. As temperature was increased motion and spin-lattice relaxation made increasing contributions to Tm. Near 100 K spin-lattice relaxation dominated Tm. PMID- 16343959 TI - Synchronized detection of minute electrical currents with MRI using Lorentz effect imaging. AB - The blood oxygenation level-dependent (BOLD) effect is the most commonly used contrast mechanism in functional magnetic resonance imaging (fMRI), due to its relatively high spatial resolution and sensitivity. However, the ability of BOLD fMRI to accurately localize neuronal activation in space and time is limited by the inherent hemodynamic modulation. There is hence a need to develop alternative MRI methods that can directly image neuroelectric activity, thereby achieving both a high temporal resolution and spatial specificity as compared to conventional BOLD fMRI. In this paper, we extend the Lorentz effect imaging technique, which can detect spatially incoherent yet temporally synchronized minute electrical activity in a strong magnetic field, and demonstrate its feasibility for imaging randomly oriented electrical currents on the order of microamperes with a temporal resolution on the order of milliseconds in gel phantoms. This constitutes a promising step towards its application to direct imaging of neuroelectric activity in vivo, which has the same order of current density and temporal synchrony. PMID- 16343960 TI - The hydration structure of 18-crown-6/K+ complex as studied by Monte Carlo simulation using ab initio fitted potential. AB - The intermolecular potential between a 18-crown-6/K+ complex and a water molecule is derived from 1200 energy points obtained from quantum chemical calculations using the 6-31G** basis set. The ab initio fitted potential was then applied to study the structural properties of the complex in an aqueous solution using the Monte Carlo simulation method. The radial distribution function (RDF) centered at K+ to the oxygen atom of water shows a sharp first peak at 2.88 A. The corresponding coordination number, integrated up to the first minimum at 3.76 A, is 2 water molecules. The results indicate clearly that the 18-crown-6/K+ complex was solvated by the two nearest neighbors, one above and other below the ligand's plane. Evaluation was focused on the precise position and orientation of the two water molecules. It was found that the oxygen atoms of the two nearest neighbors bind to the K+ while their hydrogen atoms rotate freely around the vector perpendicular to the ligand's molecular plane. PMID- 16343961 TI - Acute exacerbation of chronic bronchitis: need for an evidence-based approach. AB - Acute exacerbations of chronic bronchitis (AECB) can be classified into three levels according to severity: (1) home treatment sufficient; (2) hospitalisation required; (3) hospitalisation in the presence of respiratory failure. This evidence-based classification is useful in ranking the clinical relevance of the episode and its outcome, and makes it possible to define the clinical history, clinical evaluation and diagnostic procedures of an exacerbation. Treatment guidelines vary according to severity, but they are essentially based on appropriate bronchodilator therapy (beta(2) agonists and/or anticholinergics, corticosteroids and antibiotics selected according to the local bacterial resistance pattern). It is important that cases requiring management in an intermediate/special respiratory care unit or intensive care unit (ICU) be identified. This is the stage where oxygen therapy and ventilatory support become particularly important. As first choice, they should be non-invasive, saving intubation and invasive ventilatory support for most severe cases characterised by severe acidemia and hypercapnia. We identify the optimal criteria for hospital discharge and follow-up of patients with AECB. In view of the chronic nature of the underlying disease, a correct follow-up is essential to avoid frequent and repeated relapses. PMID- 16343962 TI - Effect of normal and waxy maize starch on growth, food utilization and hepatic glucose metabolism in European sea bass (Dicentrarchus labrax) juveniles. AB - We determined the effect of dietary starch on growth performance and feed utilization in European sea bass juveniles. Data on the dietary regulation of key hepatic enzymes of the glycolytic, gluconeogenic, lipogenic and amino acid metabolic pathways (hexokinase, HK; glucokinase, GK; pyruvate kinase, PK; fructose-1,6-bisphosphatase, FBPase; glucose-6-phosphatase, G6Pase; glucose-6 phosphate dehydrogenase, G6PD; alanine aminotransferase, ALAT; aspartate aminotransferase, ASAT and glutamate dehydrogenase, GDH) were also measured. Five isonitrogenous (48% crude protein) and isolipidic (14% crude lipids) diets were formulated to contain 10% normal starch (diet NS10), 10% waxy starch (diet WS10), 20% normal starch (diet NS20), 20% waxy starch (diet WS20) or no starch (control diet). Another diet was formulated with no carbohydrate, and contained 68% crude protein and 14% crude lipids (diet HP). Each experimental diet was fed to triplicate groups of 30 fish (initial weight: 23.3 g) on an equivalent feeding scheme for 12 weeks. The best growth performance and feed efficiency were achieved with fish fed the HP diet. Neither the level nor the nature of starch had measurable effects on growth performance of sea bass juveniles. Digestibility of starch was higher with waxy starch and decreased with increasing levels of starch in the diet. Whole-body composition and plasma metabolites, mainly glycemia, were not affected by the level and nature of the dietary starch. Data on enzyme activities suggest that dietary carbohydrates significantly improve protein utilization associated with increased glycolytic enzyme activities (GK and PK), as well as decreased gluconeogenic (FBPase) and amino acid catabolic (GDH) enzyme activities. The nature of dietary carbohydrates tested had little influence on performance criteria. PMID- 16343963 TI - Antioxidants in the egg yolk of a wild passerine: differences between breeding seasons. AB - Carotenoids and fat-soluble vitamins play crucial roles in several physiological processes. Yolk carotenoid composition may be influenced by the bird's dietary intake and by the possible discrimination during carotenoid metabolism. Information regarding the pigment composition of passerine eggs is very limited. In the present 2-year study, we determined the carotenoid and vitamin concentrations and the percentage profile of carotenoid components in collared flycatcher (Ficedula albicollis) eggs. The major carotenoid was lutein in both years, followed by cis-lutein, zeaxanthin and beta-carotene as other significant components. Carotenoid concentration was higher in 2000 than in 2001. As caterpillar abundance and ambient temperature was lower in 2001 than in 2000, we suggest that the decreased availability of carotenoid-rich resources and the elevated antioxidant demand of the female restricted the carotenoid allocation to the eggs. Our study is the first to indicate that the concentration and profile of yolk carotenoids may considerably vary in the same wild bird population between breeding seasons with different environmental conditions. Compared to other birds, the yolk of this species was relatively rich in vitamins A and E. Vitamin E concentration was positively correlated with total carotenoid concentration, which may be beneficial for the nestlings as these antioxidants synergistically provide better protection. PMID- 16343964 TI - Apoptotic activity of frog Bombina maxima skin albumin. AB - Albumin, the most abundant protein components of blood plasma, is synthesized and secreted by liver cells in vertebrates. Recently, it was demonstrated that frog Bombina maxima albumin is also expressed in skin. Both B. maxima albumins from skin and serum (BmA-skin and BmA-serum) have similar biochemical characteristics except that the former contains haem b. Present studies showed that BmA-skin exhibited cytotoxic activity on H9 and C8166 cells. Pretreated with hemin to induce erythroid differentiation, K562 cells lost their resistance to cytotoxicity of BmA-skin. After treating cells with BmA-skin for 48 h, 50 percentage cytotoxic concentrations (CC(50)) of BmA-skin on H9, C8166 and hemin treated K562 cells were 1.31+/-0.09, 1.59+/-0.08 and 2.28+/-0.06 microM, respectively. The cell death induced by BmA-skin was mediated by apoptosis of the tested cell lines, as demonstrated by nuclear morphological changes, DNA fragmentation and DNA hypodiploidy of apoptosis cells. At BmA-skin concentration of 2 microM, 27.3%, 19.7% and 17.8% of H9, C8166 and hemin-treated K562 cells were found to be apoptotic. In contrast, BmA-serum possessed no cytotoxic and apoptosis-inducing activity on all the cell lines tested, even with concentration used up to 15 microM. These results indicated that bound haem b in BmA-skin contributed significantly to its cytotoxic and apoptosis-inducing activity on the cell lines assayed. PMID- 16343965 TI - Characterization of the AMP-activated protein kinase pathway in chickens. AB - In mammals, AMP-activated protein kinase (AMPK) is involved in the regulation of cellular energy homeostasis and, on the whole animal level, in regulating energy balance and food intake. Because the chicken is a valuable experimental animal model and considering that a first draft of the chicken genome sequence has recently been completed, we were interested in verifying the genetic basis for the LKB1/AMPK pathway in chickens. We identified distinct gene homologues for AMPK alpha, beta and gamma subunits and for LKB1, MO25 and STRAD. Analysis of gene expression by RT-PCR showed that liver, brain, kidney, spleen, pancreas, duodenum, abdominal fat and hypothalamus from 3 wk-old broiler chickens preferentially expressed AMPK alpha-1, beta-2 and gamma-1 subunit isoforms. Heart predominantly expressed alpha-2, beta-2 and gamma-1, whereas skeletal muscle expressed alpha-2, beta-2 and gamma-3 preferentially. Moreover, the AMPK gamma-3 gene was only expressed in heart and skeletal muscle. Genes encoding LKB1, MO25 alpha, MO25 beta, and STRAD beta were expressed in all examined tissues, whereas STRAD alpha was expressed exclusively in brain, hypothalamus, heart and skeletal muscle. Alterations in energy status (fasting and refeeding) produced little significant change in AMPK subunit gene transcription. We also determined the level of phosphorylated (active) AMPK in different tissues and in different states of energy balance. Immunocytochemical analysis of the chicken hypothalamus showed that activated AMPK was present in hypothalamic nuclei involved in regulation of food intake and energy balance. Together, these findings suggest a functional LKB1/AMPK pathway exists in chickens similar to that observed in mammals. PMID- 16343966 TI - delta-Aminolevulinate dehydratase (ALAD) porphyria: the first case in North America with two novel ALAD mutations. AB - The molecular basis of the enzymatic defect responsible for delta-aminolevulinate dehydratase (ALAD) porphyria (ADP) was investigated in a 14-year-old male who presented clinical and laboratory findings typical of ADP. Nucleotide sequence analysis of ALAD cDNAs from the proband revealed two novel mutations, a 265G to A base transition (C1) and a 394C to T base transition (C2), resulting in amino acid substitutions, Glu89Lys and Cys132Arg, respectively. Both mutations were present within exon 5 of the ALAD gene, and appeared to influence the binding of zinc to the enzyme which is essential for enzyme activity. It was found that the C1 mutation was inherited from his father, while the C2 mutation was from his mother. Expression of these mutant ALAD cDNAs in Chinese hamster ovary cells produced normal ALAD mRNA levels, but markedly decreased ALAD protein and enzyme activity. These results suggest that the combination of the two aberrant ALADs with little enzyme activity accounts for the markedly decreased ALAD activity observed in the proband. This case represents the molecular analysis of the ALAD gene defects in the first case of ADP identified in North America, who is a compound heterozygote for two novel ALAD gene defects. PMID- 16343967 TI - Molecular, biochemical, and genetic characterization of a female patient with Lesch-Nyhan disease. AB - Lesch-Nyhan disease (LND) is a rare X-linked recessive disorder caused by virtually complete deficiency of activity of the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT; EC 2.4.2.8). Human HPRT is encoded by a single structural gene located on the long arm of the X-chromosome (Xq26). The classical LND phenotype occurs almost exclusively in males, manifested in excessive purine production and characteristic neurological manifestations, including compulsive self-mutilation, choreoathetosis, spasticity, and occasionally developmental delay. Heterozygous females are usually phenotypically normal, due to the random inactivation of the X chromosome (Lyonization mechanism). However, six females were reported to be affected with the full biochemical and clinical manifestations of LND. All these cases were heterozygous for an HPRT mutation. Absence of transcription of the normal HPRT allele was attributed in all of them to non-random inactivation of the X chromosome carrying the normal allele. Here we describe an additional LND female, who presented with acute renal failure at the age of two months, in whom absence of transcription of the two HPRT alleles occurred due to as yet undescribed mechanism in LND females: the transcription of one HPRT allele was blocked due to a de novo X chromosome autosome translocation 46,XX,t(X:2)(q26:p25), with a breaking point encompassing the HPRT gene locus, whereas the transcription of the normal allele was inhibited due to non-random inactivation of the second X-chromosome. Cultured fibroblasts from this patient exhibited the biochemical alterations in purine nucleotide metabolism characteristic of male LND fibroblasts. PMID- 16343968 TI - Cerebral 1H MR spectroscopy showing elevation of brain guanidinoacetate in argininosuccinate lyase deficiency. AB - MR spectroscopy in a patient with argininosuccinate lyase deficiency revealed elevated cerebral guanidinoacetate signals, indicating that the phenomenon of increased levels of this compound in brain tissue is not limited to creatine deficiencies. PMID- 16343969 TI - Reduced folate carrier polymorphisms and neural tube defect risk. AB - The reduced folate carrier (RFCI) is essential for folate transport into cells. Low folate is an important cause of neural tube defects (NTDs), and a single nucleotide polymorphism (H27R) (80G-->A) in the RFCI gene has been reported to be a NTD risk factor. We investigated H27R and a 61 bp tandem repeat polymorphism as potential risk factors for NTDs, using a large homogeneous Irish population by case/control comparison, log-linear analysis, and transmission disequilibrium testing. No association was found between NTDs and H27R in mothers [p = 0.23, odds ratio (OR) 0.87, 95% confidence interval (CI) 0.69-1.09], fathers (p = 0.11, OR 0.83, 95% CI 0.66-1.04), or cases (p = 0.36, OR 0.9, 95% CI 0.72-1.12) when compared to controls or through log-linear modeling for dominant or recessive effects or with the transmission disequilibrium test for preferential allele transmission. Using log-linear models, a significant protective case effect was seen for the 61 bp polymorphism (p = 0.0039, OR 0.21, 95% CI 0.05-0.85). When analyzed by genotype, individuals homozygous for a single copy of the 61 bp sequence were underrepresented in cases as compared to controls, although these results did not reach statistical significance (p = 0.081, OR 0.5, 95% CI 0.23 1.09, goodness of fit p = 0.42). We compared the frequencies of H27R and the 61 bp polymorphism in African-Americans and American-Caucasians. The frequencies of H27R polymorphism differed significantly between the two populations (p = 0.0001). This large study does not confirm previous reports that H27R is a risk factor for NTDs. The previously unstudied 61 bp tandem repeat, however, has a possible protective NTD effect in our Irish population. This requires confirmation in other studies. PMID- 16343970 TI - Cerebral glucose metabolism in adults with early treated classic phenylketonuria. AB - Classic phenylketonuria (PKU) is characterized by severe mental retardation in untreated individuals and mild neurocognitive abnormalities in some early treated adults. The exact biochemical mechanisms underlying this neurotoxicity remain undetermined. Several theories implicate abnormal cerebral energy utilization and alterations in biochemical pathways that involve glucose metabolism. This pilot study was undertaken to investigate whether 18F-deoxyglucose positron emission tomography (PET) is an effective tool to study cerebral glucose metabolism in early treated PKU. After PET coregistration with SPGR MRI, relative glucose metabolic rates (rGMR) at the center of standard atlas positions was determined. Repeated measures MANOVA was used to assess regional metabolic differences, which were then correlated with age-specific and day-of-scan plasma phenylalanine and age. Patients with PKU in comparison to controls had decreased rGMR in cortical regions including the prefrontal, somatosensory, and visual cortices, and increased activity in subcortical regions including the striatum and limbic system. Day-of-scan phenylalanine correlated with abnormal activity in subcortical structures, and older age was associated with decreased activity in the prefrontal and visual cortices. The clinical significance of these abnormalities of glucose metabolism in specific areas of the brain remains unknown. PMID- 16343971 TI - Prenatal diagnosis for CDG Ia based on post-mortem molecular study of Guthrie card. PMID- 16343972 TI - Model predictive glycaemic regulation in critical illness using insulin and nutrition input: a pilot study. AB - Stress-induced hyperglycaemia is prevalent in intensive care, impairing the immune response. Nutritional support regimes with high glucose content further exacerbate the problem. Tight glucose control has been shown to reduce mortality by up to 43% if levels are kept below 6.1 mmol/L. This research develops a control algorithm with insulin and nutritional inputs for targeted glucose control in the critically ill. Ethics approval for this research was granted by the Canterbury Ethics Committee. Proof-of-concept clinical pilot trials were conducted on intubated, insulin-dependent Christchurch ICU patients (n=7) on constant nutritional support. A target 10-15% reduction in glucose level per hour for a desired glucose level of 4-6 mmol/L was set. 43% and 91% of glucose targets were achieved within +/-5 and +/-20%, respectively. The mean error was 8.9% (0.5 mmol/L), with an absolute range [0, 2.9] mmol/L. End glucose levels were 40% lower compared to initial values. All large target errors are attributable to sudden changes in patient physiology at low glucose values, rather than systemic deficiencies. Target errors are consistent with and explainable by published sensor error distributions. The results show that intensive model-based glucose management with nutrition control reduced absolute glucose levels progressively while reducing the severity of glycaemic fluctuation even with significant inter patient variability and time-varying physiological condition. Trials spanning longer periods of time are in development to verify the short-term pilot studies performed and to test the adaptability of the controller. Clinically, these results indicate potential in clinical use to reduce ICU mortality as well as reduce risk of severe complications. PMID- 16343973 TI - "The walls were so damp and cold" fuel poverty and ill health in Northern Ireland: results from a housing intervention. AB - This article reports the findings from an evaluation of a fuel poverty programme in the Armagh and Dungannon Health Action Zone in Northern Ireland. Focusing on a rural community, it adds to the debate surrounding the hidden nature of rural fuel poverty. As part of the programme, energy efficiency measures, including some central heating systems, were installed in 54 homes. Surveys were conducted both pre and post intervention and analysed to assess any changes. The programme demonstrated that energy efficiency intervention can lead to improvements in health and well being, increased comfort levels in the home and a reduction in the use of health services, therefore having potential cost savings for the NHS. Some households, however, remain in fuel poverty after having full central heating installed, reflecting the significant contribution of low income on the production of fuel poverty. The article concludes by suggesting that interventions in this area require commitment from multiple sectors of society, including health professionals and local communities. PMID- 16343974 TI - Severe acute respiratory syndrome coronavirus membrane protein interacts with nucleocapsid protein mostly through their carboxyl termini by electrostatic attraction. AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) membrane protein is an abundant virion protein, and its interaction with the nucleocapsid protein is crucial for viral assembly and morphogenesis. Although the interacting region in the nucleocapsid protein was mapped to residues 168-208, the interacting region in the membrane protein and the interaction nature are still unclear. In this work, by using yeast two-hybrid and surface plasmon resonance techniques, the residues 197-221 of the membrane protein and the residues 351-422 of the nucleocapsid protein were determined to be involved in their interaction. Sequence analysis revealed that these two fragments are highly charged at neutral pH, suggesting that their interaction may be of ionic nature. Kinetic assays indicated that the endodomain (aa102-221) of the membrane protein interacts with the nucleocapsid protein with high affinity (K(D)=0.55+/-0.04 microM), however, this interaction could be weakened greatly by acidification, higher salt concentration (400 mM NaCl) and divalent cation (50 mM Ca2+), which suggests that electrostatic attraction might play an important role in this interaction. In addition, it is noted that two highly conserved amino acids (L218 and L219) in the membrane protein are not involved in this interaction. Here, we show that electrostatic interactions between the carboxyl termini of SARS-CoV membrane protein and nucleocapsid protein largely mediate the interaction of these two proteins. These results might facilitate therapeutic strategies aiming at the disruption of the association between SARS-CoV membrane and nucleocapsid proteins. PMID- 16343975 TI - Molecular cloning and identification of mouse Cklfsf2a and Cklfsf2b, two homologues of human CKLFSF2. AB - Human chemokine-like factor superfamily (CKLFSF) is a novel gene family comprising CKLF and CKLFSF1-8. Among them, CKLFSF2 is highly expressed in testis and may play important roles in male reproduction. Besides, it is very active during evolution and has two counterparts in mouse. For further study, we cloned the two mouse genes by EST assembly and RT-PCR methods and designated them as mouse Cklfsf2a and Cklfsf2b. Their predicted open-reading frames (ORFs) that encode 169 and 210 amino acids, respectively, were obtained; and their predicted full-length molecular sizes that are approximately 1.2 kb for mCklfsf2a and 0.9 kb for mCklfsf2b were confirmed by Northern blot analysis. Mouse Cklfsf2a and Cklfsf2b show similarities with human CKLFSF2 in the expression patterns that are abundant in testis, hematopoietic and immune tissues; as well as in the chromosome localizations that neighbor CKLFSF1 and 3. Their putative protein products have 47.6 and 45.5% identities with hCKLFSF2, respectively; both of them contain four potential transmembrane regions and MARVEL domains, which are also similar with hCKLFSF2. Functionally, they all can affect the transcriptional activity of androgen receptor in PC-3 and HeLa cells, but mCklfsf2a is a repressor while mCklfsf2b and hCKLFSF2 are enhancers. Taken together, we conclude that mouse Cklfsf2a and Cklfsf2b are two homologues of human CKLFSF2. Studies on them would provide much help in further investigation of the latter. PMID- 16343976 TI - The neurotrophin receptor TrkB is colocalized to mitochondrial membranes. AB - The transmembrane tyrosine-specific protein kinase TrkB has been shown to serve as a receptor for the neurotrophic factors BDNF and NT-4. Neurotrophin binding to TrkB isoformes mediates many intracellular signaling pathways, including calcium signalling. Two truncated isoforms of the receptor, lacking the tyrosine kinase activity, signal through a yet unknown pathway. Specific signals modulate the surface expression of TrkB, which is localized in considerable amounts in intracellular pools. These intracellular pools has not been specified so far. We therefore investigated the intracellular distribution of TrkB by colocalisation studies. In contrast to the unspecific neurotrophin receptor NGFRp75, TrkB immunohistochemistry showed a staining pattern very similar to mitochondrial stainings in adult human skeletal muscle fibers. Immunofluorescence techniques revealed in different types of permeabilized cells that TrkB is bound to mitochondrial membranes. This observation was confirmed on isolated astrocyte mitoplasts. Colocalisation of the TrkB ligand NT-4 and the specific mitochondrial marker cytochrome c oxidase was also observed. Western blot analysis of isolated mitochondria from rat brain and skeletal muscle verified that a truncated isoform of TrkB is present in both, brain and muscle mitochondria, and full-length TrkB is additionally present in brain mitochondria. Our results imply that neurotrophins can be stored in mitochondria and possibly act as signalling molecules on mitochondria. PMID- 16343977 TI - The grape-derived polyphenol resveratrol differentially affects epidermal and platelet-derived growth factor signaling in human liver myofibroblasts. AB - The grape-derived polyphenol resveratrol is anti-proliferative for human liver myofibroblasts, which may be beneficial for the treatment of liver fibrosis. However, its mechanism of action is ill understood. Here, we have studied how resveratrol interfered with signaling pathways used by epidermal or platelet derived growth factors to induce the proliferation of these cells. We found that resveratrol inhibited epidermal growth factor or platelet-derived growth factor induced DNA synthesis. Resveratrol did not, however, decrease epidermal growth factor receptor autophosphorylation or activation of extracellular regulated kinases, but strongly inhibited the phosphorylation of Akt and of its substrate forkhead related transcription factor. This suggested that resveratrol inhibited epidermal growth factor-induced mitogenic signaling through inhibition of the phosphatidylinositol 3-kinase /Akt pathway. The phosphatidylinositol 3-kinase inhibitor LY 294002, also, inhibited epidermal growth factor-dependent DNA synthesis and Akt phosphorylation but did not decrease extracellular regulated kinases phosphorylation. In contrast, resveratrol inhibited platelet-derived growth factor-stimulated receptor autophosphorylation and every subsequent signaling step. Resveratrol did not directly inhibit phosphatidylinositol 3 kinase activity measured on immunoprecipitates from epidermal growth factor stimulated myofibroblasts, but it strongly reduced the autophosphorylation of the phosphatidylinositol 3-kinase downstream target phospho-inositide-dependent kinase-1 that phosphorylates Akt. We, thus, show that resveratrol has growth factor-specific effects: it inhibits platelet-derived growth factor signaling via reduced receptor activation, whereas it reduces epidermal growth factor-dependent DNA synthesis via inhibition of the phosphatidylinositol 3-kinase/Akt pathway, possibly through inhibition of phospho-inositide-dependent kinase-1 activity. PMID- 16343978 TI - Differential expression of MHC class II molecules in highly metastatic breast cancer cells is mediated by the regulation of the CIITA transcription Implication of CIITA in tumor and metastasis development. AB - We analyzed the differential gene expression between variants of MDA-MB-435 human breast cancer cell line that share an identical genetic background but have different metastatic ability. The major histocompatibility complex class II was found down-regulated in highly metastatic cells and correlated with MHC transactivator (CIITA) expression. Constitutive CIITA expression observed in poorly metastatic is driven by promoters III and IV of CIITA gene. Conversely, both promoters were ineffective in highly metastatic cells. The MHC class II and CIITA expression was restored in these cells upon stimulation with IFNgamma or by the treatment with a hypomethylating agent. Both treatments induced USF-1 and IRF binding complexes to promoter IV but only IFNgamma induced the binding of 435 Lung2 nuclear proteins to an ARE-1 site at the promoter III. Neither Southern blot nor bisulfite sequencing of promoter IV demonstrated strong hypermethylation of this promoter at the IFNgamma-responsive elements such as GAS, E-box or IRF-1. We suggest that partial or hemimethylation of promoter IV is sufficient to silence the CIITA expression in highly metastatic cells and that this epigenetic mechanism is responsible for the lack of MHC-II expression. Forced CIITA expression restored the MHC-II antigen expression in 435-Lung2 cells and abrogates spontaneous lung metastasis in both SCID and nude mice but also affected the tumorigenicity in nude mice. The increase in NK cell infiltration in nude mice bearing CIITA-tumors correlated with sign of tumor cell apoptosis and the increase in the number of NK cells in the spleens, suggesting that NK cells might be responsible for the observed antitumor activity. PMID- 16343979 TI - Translation initiation factors: a weak link in plant RNA virus infection. AB - Recessive resistance genes against plant viruses have been recognized for a long time but their molecular nature has only recently been linked to components of the eukaryotic translation initiation complex. Translation initiation factors, and particularly the eIF4E and eIF4G protein families, were found to be essential determinants in the outcome of RNA virus infections. Viruses affected by these genes belong mainly to potyviruses; natural viral resistance mechanisms as well as mutagenesis analysis in Arabidopsis all converged to identify the same set of translation initiation factors. Their role in plant resistance against RNA viruses remains to be elucidated. Although the interaction with the protein synthesis machinery is probably a key element for successful RNA virus infection, other possible mechanisms will also be discussed. PMID- 16343980 TI - Degradation of negative regulators: a common theme in hormone and light signaling networks? AB - Signal transduction pathways often modulate both positively and negatively acting components to optimize the efficiency of a signal. Recent results have shown that plants make extensive use of regulated proteolysis to modulate signal transduction pathways. An emerging theme from hormone (e.g. auxin and gibberellin) and light signaling pathways is signal or stimulus-induced degradation of negative regulators to optimize plant growth and development. PMID- 16343981 TI - Taeniasis/cysticercosis and echinococcosis with focus on Asia and the Pacific. Proceedings of the 5th International Symposium, Asahikawa, Japan, 2005. PMID- 16343982 TI - Mathematical modeling of Echinococcus multilocularis transmission. AB - A mathematical model for the transmission cycle of Echinococcus multilocularis would be useful for estimating its prevalence, and the model simulation can be instrumental in designing various control strategies. This review focuses on the epidemiological factors in the E. multilocularis transmission cycle and the recent advances of mathematical models for E. multilocularis transmission. PMID- 16343983 TI - A checklist of parasites recorded on sticklebacks (Actinopterygii: Gasterosteidae) from Poland. AB - Parasites of the three-spined stickleback, Gasterosteus aculeatus (L.), and the nine-spined stickleback, Pungitius pungitius (L.), from brackish and freshwater habitats in Poland are listed. The following list was compiled from published records and own long-term studies. Parasites are listed alphabetically in meaningful groups of parasites with notes about their location on the host and distribution in Poland with references. A total of 51 species (taxa) of parasites were recorded on G. aculeatus (3 Apicomplexa, 1 Microsporidia, 3 Peritrichia, 1 Myxozoa, 7 Digenea, 2 Monogenea, 10 Cestoda, 10 Nematoda, 7 Acanthocephala, 3 Copepoda, 1 Branchiura, 2 Hirudinea, 1 Bivalvia) and 16 parasites on P. pungitius (1 Apicomplexa, 1 Microsporidia, 2 Peritrichia, 4 Digenea, 1 Monogenea, 2 Cestoda, 2 Nematoda, 1 Copepoda, 1 Branchiura, 1 Bivalvia). PMID- 16343984 TI - Antileishmanial activity of Eugenol-rich essential oil from Ocimum gratissimum. AB - Leishmaniasis is a group of diseases with a large spectrum of clinical manifestations caused by protozoans of the genus Leishmania. Here we demonstrate the leishmanicidal activity of the essential oil of Ocimum gratissimum as well as its main constituent, eugenol. The eugenol-rich essential oil of O. gratissimum progressively inhibited Leishmania amazonensis growth at concentrations ranging from 100 to 1000 microg/ml. The IC50 (sub-inhibitory concentration) of the essential oil for promastigotes and amastigotes were respectively 135 and 100 microg/ml and the IC50 of eugenol was 80 microg/ml for promastigote forms. L. amazonensis exposed to essential oil at concentrations corresponding to IC50 for promastigotes and for amastigotes underwent considerable ultrastructural alterations, as shown by transmission electron microscopy. Two or more nuclei or flagella were observed in 31% and 23.3% of treated amastigote and promastigote forms, respectively, suggesting interference in cell division. Considerable mitochondrial swelling was observed in essential oil-treated promastigotes and amastigotes, which had the inner mitochondrial membrane altered, with a significant increase in the number of cristae; in some amastigotes the mitochondrial matrix became less electron-dense. The minimum inhibitory concentration for both promastigotes and amastigotes was 150 microg/ml. Pretreatment of mouse peritoneal macrophages with 100 and 150 microg/ml essential oil reduced the indices of association between promastigotes and the macrophages, followed by increased in nitric oxide production by the infected macrophages. The essential oil showed no cytototoxic effects against mammalian cells. This set of results suggests that O. gratissimum essential oil and its compounds could be used as sources for new antileishmanial drugs. PMID- 16343985 TI - WHO classification of alveolar echinococcosis: principles and application. AB - Alveolar echinococcosis is caused by the larval stage of the fox tapeworm (Echinococcus multilocularis) and is frequently diagnosed as a space occupying lesion in the liver. The growth pattern resembles that of a malignant tumor with infiltration throughout the liver, spreading into neighbouring organs and metastases formation in distant organs. Thus, one of the prevailing differential diagnoses is liver cancer. Guided by the Tumor-Node-Metastasis (TNM) system of liver cancer, the European Network for Concerted Surveillance of Alveolar Echinococcosis and the WHO Informal Working Group on Echinococcosis proposed a clinical classification for alveolar echinococcosis. It was designated as PNM system (P = parasitic mass in the liver, N = involvement of neighbouring organs, and M = metastasis). As for TNM in oncology, single PNM categories were combined into four stages, I to IV. The system was developed by a retrospective analysis of 97 patients' records from two treatment centers (Besancon/France and Ulm/Germany). Recently, this WHO classification was applied to 222 patients in 4 clinical centers around the world (Besancon/France, n = 26; Urumqi/China, n = 46; Sapporo/Japan, n = 58; and Ulm/Germany, n = 92). All patients could be classified who had been diagnosed in the period from January 1998 to June 2005. The stage grouping indicated center differences, but appeared to segregate patients according to various treatment regimens. The WHO classification not only serves as a tool for the international standardization of disease manifestation but also aids to evaluate the outcome of a chosen diagnostic and treatment procedure in different treatment centers in Europe and Asia. PMID- 16343986 TI - Veterinary public health activities at FAO: cysticercosis and echinococcosis. AB - In many developing and transition countries, parasitic zoonoses such as cysticercosis and echinococcosis cause serious human suffering and considerable losses in agricultural and human productivity, thus posing a significant hindrance to their development. Although, effective and reliable tools for the diagnosis, prevention and control of parasitic zoonoses are now available, their implementation has not always been successful in many countries. This is primarily due to the lack of awareness on the presence or impact of the causing parasites (Taenia saginata, Taenia solium and Echinococcus spp.). In addition, often the needed intersectorial cooperation, resource management and political commitment for their control are (also) absent. FAO's regular programme has established a global network of professionals directly involved in zoonotic and food-borne diseases. The network provides a basic framework for the spread of information related to the diagnosis, prevention and control of major zoonotic diseases including cysticercosis and echinococcosis. PMID- 16343987 TI - The molecular mechanisms of larval cestode development: first steps into an unknown world. AB - Several hundred million years ago, the free-living ancestors of all extant helminth parasites decided to colonize entirely new habitats, the bodies of other metazoan animals. As a consequence of the resulting adaptation processes, they evolved highly complex life-cycles in which many developmental transitions were initiated and controlled by host-derived signals. Understanding the molecular basis of the original developmental mechanisms, and the modifications that occurred during co-evolution with the host, is not only fundamental to our understanding of parasitism but also highly relevant for the design of anti parasitic drugs and vaccines. In the past several years, molecular investigations on parasitic nematode and trematode development have made considerable progress and, supported by respective genome sequencing projects and emerging methods of genetic manipulation, will be a flourishing field in the years to come. We consider it time that corresponding studies are also pushed for the third large group of parasitic helminths, the cestodes. Here, we review the first experimental steps into that area, which have been undertaken recently. We report on cestode genomics, the identification of signaling factors associated with larval development, and the establishment as well as improvement of in vitro cultivation systems by which cestode life-cycles can be studied in the laboratory. PMID- 16343988 TI - Health practitioners' and health planners' information needs and seeking behavior for decision making in Uganda. AB - BACKGROUND: Access to reliable information is the most cost-effective and achievable strategy for sustainable improvement in health care. While several studies have described practitioners' information seeking behavior in developed countries, literature from developing countries is lacking. The aims of the study were: (i) to determine the most influential type of information for health workers' and planners' decision making; (ii) to establish the practitioners' evaluation of the availability and quality of this important information; and (iii) to establish the most commonly used/accessible sources of the information relevant for decision making in Uganda. METHODS: Self-administered questionnaires were sent to 610 health workers, planners, and administrators at the national, district and health facility levels in four districts in Uganda. Respondents were reminded three times, after which non-response was registered. Data were entered, cleaned and analysed using SPSS version 12.0. Logistic regression analysis was used to test for differences in responses. RESULTS: The response rate was 67.7% (413). The respondents indicated that personal experience (79%), discussion with colleagues (76%) and national policy and treatment guidelines (75%) were most influential when making decisions in health care and planning. They reported that most of the epidemiological information was available and of relatively good quality but there was lack of information about distribution of benefit, segregated demographic data, and social values. The most often used sources of information included; discussions with colleagues (89%), doctors' statements (85%) and text books (77%). The least frequently used sources were the internet and the library. This varied with respondents' designation and region of origin. CONCLUSIONS: Health planners and practitioners lack some of the information relevant for decision making. They tend to rely on the national policy and treatment guidelines, discussion with colleagues, and personal experience. Most frequently used sources of information are similar to previous studies. The variation in the frequency of use of the different sources of research information necessitates research to understand the barriers and careful planning of health information delivery to ensure equitable access. PMID- 16343989 TI - The role of trehalose dimycolate (cord factor) on morphology of virulent M. tuberculosis in vitro. AB - SETTING: M. tuberculosis (MTB) lose virulence during prolonged culture on artificial media. This loss of virulence is associated with a change in colony morphology. Several studies suggested that trehalose 6,6' dimycolate (TDM or cord factor), contributes to colony morphology. OBJECTIVE: To investigate the role of TDM in colony morphology of MTB using clinical isolates selected to have colony morphology typical of virulent or attenuated organisms. DESIGN: Use immunohistochemical and physical chemical methods to assess the presence and distribution of TDM in rapidly growing pellicles of MTB. RESULTS: TDM forms an insoluble crystalline monolayer at the air-water interfaces that is more rigid than that formed by any other biologic amphiphile and is strong enough to support a spreading pellicle of MTB. The surface of young pellicles of the isolate with virulent morphology displayed the regular linear pattern characteristic of monolayers of TDM. TDM was also identified in the open spaces of pellicles of MTB by immunohistochemistry. MTB with morphology of attenuated organisms had neither of these properties. CONCLUSION: These data suggest that the characteristic morphology of colonies of virulent MTB is due to TDM released from the surface of the organisms. PMID- 16343990 TI - Rheohaemapheresis of ophthalmological diseases and diseases of the microcirculation. AB - Blood rheology was considered to be of limited clinical importance, until extracorporeal technologies enabling the treatment of cellular and plasmatic hyperviscosity syndromes were introduced. However, a wide range of applications, mainly referring to rheologically determined diseases of the microcirculation exists but has so far hardly been taken into consideration. The extension of indications was due to modern technical developments leading to different approaches of secondary separation such as precipitation, ad- or absorption and filtration. Based on 18 years of experience with different separation technologies the combination of some centrifugal devices applied for cell--plasma separation with secondary filtration (Rheohemapheresis) appears to be the most efficacious and economical approach for such extracorporeal treatments. The sequence and frequency of rheohaemapheresis treatments depends on the measurement of rheological measurements in combination with clinical--chemical data which are related to the distribution kinetics and synthesis rate of both blood cells and plasma proteins. Standardised treatment protocols proved the efficacy of initial therapies and were applied for controlled trials whereas long term therapies may need more flexible treatment approaches. So far an increasing experience exists for the treatment of ophthalmologic diseases, otologic disorders, diabetic complications and cardiological diseases. Rheohemapheresis was shown to be a safe treatment approach, if a careful risk assessment prior to the initiation of the therapies is performed. The treatment cannot cure diseases but enables a substantial improvement of the quality of life in patients without treatment alternatives. PMID- 16343991 TI - Investigation of the role of complement and complement receptors in the modulation of B cell activation by a Paracoccidioides brasiliensis cell wall fraction. AB - F1 fraction from Paracoccidioides brasiliensis is a potent activator of the complement system. Considering that complement receptors CR1 and CR2 are involved in the regulation of B cell response, we evaluated the in vitro effect of the F1 in the activation of B lymphocytes, as well as the participation of complement receptors in this process. Murine splenocytes were cultured in order to evaluate the expression of CD40, CD45RB and CD69 on B lymphocyte, and IgG and IgM were quantified in the culture supernatant. F1 participated in the activation of B cells, showing a positive modulation effect on all markers analyzed. An increase in the production of IgG was detected in the supernatants when the opsonized F1 fraction was present. Complement receptor blockade with monoclonal antibodies led to a partial reduction in immunoglobulin secretion, suggesting that these receptors, especially CR2, play a role in modulating the function of B lymphocyte stimulated with the opsonized F1 fraction. These results may contribute for a better understanding of the B cell activation and differentiation processes in response to the F1 fraction from P. brasiliensis. PMID- 16343992 TI - Altered collagen II peptides inhibited T-cell activation in rheumatoid arthritis. AB - It has been reported that collagen II (CII)-derived peptide induced T-cell activation via its amino acids responsible for T-cell receptor (TCR) recognition. In this study, three altered CII263-272 peptide ligands (APL) containing multiple substitutions of TCR contact residues were synthesized. Their roles in inhibition of T-cell activation were evaluated in peripheral blood lymphocytes (PBL) of rheumatoid arthritis (RA) in vitro. It was shown that 41% (25/61) of RA patients were responsive to the wild-type antigenic CII263-272. In contrast, marginal or silent T-cell responses to the three APLs were found, accompanied by inhibitory effects on secretion of Th1 type cytokines and expression of cell surface markers, CD69 and CD25. In addition, T-cell activation induced by the wild-type antigenic CII263-272 was inhibited by all the three APLs in a dose-dependent manner. It is demonstrated that APLs with substitutions of TCR contact residues are capable of down-regulating T-cell responses in PBLs of RA, suggesting that the CII-derived APLs are potentially therapeutic in RA. PMID- 16343993 TI - Development of cell-based tuberculosis vaccines: genetically modified dendritic cell vaccine is a much more potent activator of CD4 and CD8 T cells than peptide- or protein-loaded counterparts. AB - Genetically modified dendritic cell (DC)-based vaccines have not been explored for immunization against tuberculosis. A gene-modified DC vaccine expressing Mycobacterium tuberculosis (M.tb) antigen 85A (Ag85A) was developed by using a recombinant replication-deficient adenoviral gene transfer vector (AdAg85A). AdAg85A-transduced DC vaccine (AdAg85/DC) expressed higher levels of IL-12 and was much more immunogenic than Ag85 protein-loaded (pro/DC) or CD4/CD8 T cell peptide-loaded (pep/DC) DC vaccines. Compared to pro/DC or pep/DC, AdAg85/DC elicited a remarkably higher level of ex vivo IFN-gamma production by CD4 and CD8 T cells at weeks 2, 6, and 12 postimmunization, which was coupled with higher frequencies of antigen-specific T cells. By an in vivo CD8 or CD4 T cell cytotoxicity (CTL) assay, AdAg85/DC was shown to provoke much higher and more sustained levels of CD8 and CD4 CTL activity up to 12 weeks postimmunization. Intramuscular (im) AdAg85/DC immunization was more potent than the iv route of AdAg85/DC immunization. Such stronger immunogenicity of im AdAg85/DC vaccination was corroborated with better protection from M.tb challenge. Our results thus suggest that genetically modified DC-based TB vaccine is superior to subunit DC vaccines and has the potential for therapeutic applications. PMID- 16343994 TI - Design of noninflammatory synthetic siRNA mediating potent gene silencing in vivo. AB - Targeted silencing of disease-associated genes by synthetic short interfering RNA (siRNA) holds considerable promise as a novel therapeutic strategy. However, unmodified siRNA can be potent triggers of the innate immune response, particularly when associated with delivery vehicles that facilitate intracellular uptake. This represents a significant barrier to the therapeutic development of siRNA due to toxicity and off-target gene effects associated with this inflammatory response. Here we show that immune stimulation by synthetic siRNA can be completely abrogated by selective incorporation of 2'-O-methyl (2'OMe) uridine or guanosine nucleosides into one strand of the siRNA duplex. These noninflammatory siRNA, containing less than 20% modified nucleotides, can be readily generated without disrupting their gene-silencing activity. We show that, coupled with an effective systemic delivery vehicle, 2'OMe-modified siRNA targeting apolipoprotein B (apoB) can mediate potent silencing of its target mRNA, causing significant decreases in serum apoB and cholesterol. This is achieved at therapeutically viable siRNA doses without cytokine induction, toxicity, or off-target effects associated with the use of unmodified siRNA. This approach to siRNA design and delivery should prove widely applicable and represents an important step in advancing synthetic siRNA into a broad range of therapeutic areas. PMID- 16343995 TI - Inhibition of ocular neovascularization by hedgehog blockade. AB - Ocular neovascularization associated with proliferative diabetic retinopathy and age-related macular degeneration is the leading cause of severe visual loss in adults in developed countries. Physiological and pathological retinal angiogenesis may occur independently in postnatal life through the complex activation of pro- and antiangiogenic pathways. We report that the Sonic hedgehog (Shh) pathway is activated in the retina in animal models of retinal and choroidal neovascularization. We show that pharmacological inhibition of the Shh signaling pathway significantly reduces physiological retinal angiogenesis and inhibits pathological vascularization in both models. Under retinal hypoxic conditions, inhibition of the Shh pathway results in reduction of vascular endothelial growth factor (VEGF) level, along with that of Patched-1 (Ptch1), a canonical Shh target, thus placing Shh activation upstream of VEGF in experimental retinal neovascularization. Our data demonstrate the requirement of the Shh pathway for retinal angiogenesis and its inhibition as a potential therapeutic strategy targeting ocular neovascular disease. PMID- 16343997 TI - Vagus nerve stimulation does not lead to significant changes in body weight in patients with epilepsy. AB - BACKGROUND: Vagus nerve stimulation (VNS) is an FDA-approved treatment for medically intractable epilepsy. The effect of this therapy on body weight is unclear. VNS could cause weight loss by engaging vagal afferents from the gastrointestinal tract mediating satiety. METHODS: We performed a retrospective analysis of body weight changes over a period up to 2 years following VNS implantation. We studied 21 patients (13 M/8 F) 35 +/- 12 years old, who received a Cyberonics VNS Therapy System for medically intractable epilepsy between April 1998 and May 2004. The mean +/- SD duration of follow-up was 613.1 +/- 389.1 days. The study had 80% power with a type I error of 0.05 to detect a 5% weight change. Data were analyzed with repeated-measures ANOVA. RESULTS: Weight changes relative to baseline at 30, 60, 120, 360, 480, and 720 days were -0.17 +/- 2.33, +0.33 +/- 2.64, -0.32 +/- 3.56, +1.09 +/- 5.97, +1.06 +/- 7.47, and +0.33 +/- 3.69%, respectively. At all time points these differences failed to reach statistical significance. CONCLUSIONS: Vagus nerve stimulation with parameters typically used in the treatment of patients with epilepsy was not associated with clinically significant weight changes. A well-controlled prospective study is necessary for more precise evaluation of the effect of VNS therapy on body weight. PMID- 16343998 TI - Recurrent status epilepticus as the main feature of Hashimoto's encephalopathy. AB - Hashimoto's encephalopathy (HE) is a severe but treatable condition that rarely complicates Hashimoto's thyroiditis. Clinically it is characterized by progressive or relapsing symptoms, including tremor, myoclonus, stroke-like episodes, seizures, impairment of consciousness, and dementia. We describe a patient presenting with recurrent generalized convulsive status epilepticus (GCSE), despite antiepileptic medications, who was successfully treated with methylprednisolone. Our observation confirms that the clinical spectrum of HE at presentation is heterogeneous and diagnosis is often difficult. This case highlights the crucial importance of antithyroid antibody measurement in patients presenting with otherwise unexplained episodes of GCSE with or without adjunctive signs of encephalopathy or thyroiditis. PMID- 16343999 TI - Antioxidant properties of the mucus secreted by Laeonereis acuta (Polychaeta, Nereididae): a defense against environmental pro-oxidants? AB - Polychaeta species like Laeonereis acuta (Nereididae) usually secrete great amounts of mucus that wrap the animal inside. Taking into account that fungi action in the sediment and UV radiation acting on dissolved organic matter in the water produces reactive oxygen species (ROS) like hydrogen peroxide (H(2)O(2)), it was considered that the mucus secretion could represent an antioxidant defense against environmental ROS. Antioxidant enzymes (catalase-CAT; superoxide dismutase-SOD; glutathione peroxidase-GPx and glutathione-S-transferase-GST) and total antioxidant capacity (TOSC) were determined in worms and mucus secretion. Higher (p<0.05) CAT, GPx and TOSC values were registered in mucus samples respect worms, SOD activity was similar (p>0.05) in both kind of samples, and absence of GST activity was observed in mucus samples, suggesting absence of catalyzed phase II reactions. In assays conducted with hepatoma cell lines exposed to H(2)O(2), it was verified that: (1) mucus co-exposure significantly (p<0.05) lowered DNA damage induced by H(2)O(2); (2) ROS production was significantly (p<0.05) reduced when cells were exposed simultaneously with mucus samples and H(2)O(2) respect H(2)O(2) alone. It can be concluded that the mucus production contributes substantially to the antioxidant defense system of the worm against environmental ROS through the interception or degradation of H(2)O(2), peroxyl and hydroxyl radicals. PMID- 16344000 TI - Pharmacogenetics and cardiac ion channels. AB - Ion channels control electrical excitability in living cells. In mammalian heart, the opposing actions of Na(+) and Ca(2+) ion influx, and K(+) ion efflux, through cardiac ion channels determine the morphology and duration of action potentials in cardiac myocytes, thus controlling the heartbeat. The last decade has seen a leap in our understanding of the molecular genetic origins of inherited cardiac arrhythmia, largely through identification of mutations in cardiac ion channels and the proteins that regulate them. Further, recent advances have shown that 'acquired arrhythmias', which occur more commonly than inherited arrhythmias, arise due to a variety of environmental factors including side effects of therapeutic drugs and often have a significant genetic component. Here, we review the pharmacogenetics of cardiac ion channels-the interplay between genetic and pharmacological factors that underlie human cardiac arrhythmias. PMID- 16344002 TI - Study design and rationale of a mock phase I trial as an educational tool for clinical pharmacology residents. AB - OBJECTIVE: To invite comments and suggestions from the phase I trialists, the world over, on the design and rationale provided by us for conducting a mock phase I trial for training clinical pharmacology residents. METHODS: The trial will be conducted by clinical pharmacology residents of the Post Graduate Institute of Medical Education and Research. After a thorough evaluation of Preclinical toxicity data of a pharmaceutical product provided by a dummy pharmaceutical firm, a randomized, double-blind, placebo controlled, parallel group design with dose escalation of the product will be carried out. A single dose administration will be followed by monitoring the participants for 24 h and again at one week. After obtaining results of the previous group, the next higher dose will be administered. Evaluation of the learners will be done by two independent teachers who will mark them for actual conduct of the study and writing a report of the study. Subsequently, we will try to publish the results of the mock trial and invite further comments. CONCLUSIONS: This trial designed primarily for training purposes will be instrumental in equipping the residents with the expertise for conducting phase I trials. PMID- 16344003 TI - Action site of ketanserin enhancing baroreflex function is within the rostral ventrolateral medulla in anesthetized rats. AB - In our previous studies, it was found that ketanserin enhanced baroreflex sensitivity (BRS) in rats and this effect was not blood pressure dependent. The present work was designed to investigate the effects of ketanserin on BRS within the nucleus tractus solitarus (NTS) and the rostral ventrolateral medulla (RVLM). In anesthetized rats, BRS were evaluated by the changes in depressor action of aortic nerve stimulation and bradycardiac response to rapid pressor action of intravenous phenylephrine. It was found that bilateral injection of ketanserin (250 pmol for each side) into the NTS not only significantly increased blood pressure, but also attenuated baroreflex function. Interestingly, when ketanserin was bilaterally injected into RVLM, a significant enhancement of BRS was found and there were no modifications in basal blood pressure and heart period. The present study demonstrates that ketanserin-induced enhancement of BRS mainly occurred within the RVLM, which might contribute to the systemic effects of ketanserin on BRS. PMID- 16344004 TI - The ornithine decarboxylase gene of Trypanosoma brucei: Evidence for horizontal gene transfer from a vertebrate source. AB - Kinetoplastid protozoans in the family Trypanosomatidae are parasites, many of them responsible for serious diseases in humans and domestic animals. Ornithine decarboxlyase (ODC), a protein at the core of polyamine metabolism, is a potential target for therapies to overcome these diseases. Eukaryotic phylogenies were constructed from full-length genes for ODC to determine the origin of ODC in the kinetoplastid protozoans. The Odc genes from Trypanosoma brucei and two other African trypanosomes, T. congolense and T. vivax, clustered with Odc genes from vertebrates rather than with Odc genes from other kinetoplastids and other protozoans, making this gene a candidate for horizontal gene transfer from a vertebrate source. This result is unique to the Odc gene from the African trypanosomes as four other genes produced phylogenies consistent with the expected taxonomic relationships for the organisms. Analysis of the genomic regions around the Odc genes in Leishmania major, T. brucei, and Trypanosoma cruzi supports the hypothesis of loss of the Odc gene in the Trypanosoma lineage followed by acquisition of a new copy from a vertebrate host in the African branch of the genus. PMID- 16344005 TI - Dysfunction of lamin A triggers a DNA damage response and cellular senescence. AB - In higher eukaryotes, the nuclear lamins play an important role in maintaining the integrity of the nuclear envelope and the nucleus itself. Two recent papers show that a mutation that affects the processing of one of the nuclear lamins, lamin A, results in increased sensitivity to DNA damaging agents, an elevated DNA damage response, and a senescent phenotype. These studies underscore the role of the nuclear envelope in maintaining genomic stability and the interplay between nuclear architecture and the DNA damage response. PMID- 16344006 TI - Chronic hypoxia modulates NMDA-mediated regulation of the hypoxic ventilatory response in an amphibian, Bufo marinus. AB - This study examined whether a hypoxia-tolerant amphibian, the Cane toad, undergoes mammalian-like ventilatory acclimatisation to hypoxia (VAH) and whether chronic hypoxia (CH) alters NMDA-mediated regulation of the acute hypoxic ventilatory response (HVR). Toads were exposed to 10 days of CH (10% O2) followed by acute hypoxic breathing trials or an intra-arterial injection of NaCN. Trials were conducted before and after i.p. treatment with an NMDA-receptor channel blocker (MK801). CH blunted the acute HVR but did not alter resting breathing. MK801 did not alter resting ventilation. In control animals, MK801 augmented breathing frequency (fR) during acute hypoxia by increasing the number of breaths per episode. This effect was attenuated following CH although MK801 did enhance the number of episodes per minute during acute hypoxia. MK801 enhanced the fR response to NaCN in both groups. The results indicate that CH did not produce mammalian-like VAH (i.e. increased resting ventilation and an augmented acute HVR) but did alter MK801-sensitive regulation of breathing pattern and the acute HVR. PMID- 16344007 TI - Muscle-targeted deletion of VEGF and exercise capacity in mice. AB - Methods to study exercise are evolving from classically integrative organ approaches towards the more fundamental cellular reactions. While in vitro cellular and molecular methods are well established, only recently has in vivo molecular manipulation been widely used. This review discusses two complementary methods for determining in vivo the significance of one gene thought important to exercise: vascular endothelial growth factor (VEGF). Because VEGF deletion is embryonically lethal, its study requires conditional and/or organ-targeted strategies. We inactivated the muscle VEGF gene in two ways: PMID- 16344008 TI - Systemic oxygen transport in rats artificially selected for running endurance. AB - The relative contribution of genetic and environmental influences to individual exercise capacity is difficult to determine. Accordingly, animal models in which these influences are carefully controlled are highly useful to understand the determinants of intrinsic exercise capacity. Studies of systemic O(2) transport during maximal treadmill exercise in two diverging lines of rats artificially selected for endurance capacity showed that, at generation 7, whole body maximal O(2) uptake ((.)V(O(2)(max)) was 12% higher in high capacity (HCR) than in low capacity runners (LCR) during normoxic exercise. The difference in (.)V(O(2)(max) between HCR and LCR was larger during hypoxic exercise. Analysis of the linked O(2) conductances of the O(2) transport system showed that the higher (.)V(O(2)(max) was not due to a higher ventilatory response, a more effective pulmonary gas exchange, or an increased rate of O(2) delivery to the tissue by blood. The main reason for the higher (.)V(O(2)(max) of HCR was an increased tissue O(2) extraction, due largely to a higher tissue diffusive O(2) conductance. The enhanced tissue O(2) diffusing capacity was paralleled by an increased capillary density of a representative locomotory skeletal muscle, the gastrocnemius, in HCR. Activities of skeletal muscle oxidative enzymes citrate synthase and beta-HAD were also higher in HCR than LCR. Thus, the functional characteristics observed during exercise are consistent with the structural and biochemical changes observed in skeletal muscle that imply an enhanced capacity for muscle O(2) uptake and utilization in HCR. The results indicate that the improved (.)V(O(2)(max) is solely due to enhanced muscle O(2) extraction and utilization. However, the question arises as to whether it is possible to maintain a continually expanding capacity for O(2) extraction at the tissue level with successive generations, without a parallel improvement in the capacity to deliver O(2) to the exercising muscles. PMID- 16344009 TI - Improved solid-phase extraction and HPLC measurement of torasemide and its important metabolites. AB - Torasemide is a "loop type" diuretic drug. For pharmacokinetic studies sensitive analytic methods are essential for authentic qualitative and quantitative information. A robust, selective and sensitive HPLC method is described for the simultaneous determination of torasemide, its major metabolite M5 and its active metabolites M1 and M3 and an internal standard within 17 min. Solid-phase extraction with C(2)-cartridges was used for the clean-up of plasma samples. The chromatographic separation was carried out on a CN-column with a mobile phase consisting of perchloric acid (0.02 M, pH 2.5)/acetonitrile (90/10, v/v)). The calibration range used reached from 20 to 1000 ng/ml for all analytes. Coefficients of variation were less than 10% at every calibration point for each analyte. Plasma concentrations in samples obtained from volunteers in the course of a clinical study could be reliably measured with this method. Median maximum concentrations in plasma after a 10mg oral dose during a 24h study interval were located at 1h for torasemide, 1h for M1 and 2h for M5. Concentrations between 2226 and <20 ng/ml for torasemide, between 159 and <20 ng/ml for M1 and between 420 and <20 ng/ml for M5 were observed. PMID- 16344010 TI - High-performance liquid chromatography determination of alpha1-acid glycoprotein in small volumes of plasma from neonates. AB - In order to investigate how the alpha1-acid glycoprotein (AGP) concentrations of neonates change in response to surgical stress, a simple high-performance liquid chromatography (HPLC)-assay for the measurement of alpha1-acid glycoprotein levels was developed. A fraction containing alpha1-acid glycoprotein was isolated from the bulk of plasma protein by addition of 0.6M perchloric acid and was then analysed directly on a short PLRP-S 4000A reversed phase column column. The method was validated by analysis of pooled plasma from healthy adults both in comparison with a calibration curve and by standard additions. The procedure was able to isolate alpha1-acid glycoprotein rapidly (<30 min) and required only 50 microl of plasma. The mean extraction recovery was 79.1% (CV 6.4%). The within run precision for the analysis of three replicates of quality control sample ranged from +/-1.2 to +/-3.8% and the between-run precision was +/-6.1%. The method was linear (r(2)=0.988) over a concentration range from 6 to 100.0 mg/100 ml. The AGP levels in neonatal samples ranged from 25 to 93 mg/100 ml. PMID- 16344011 TI - Crystal structure of CTP:glycerol-3-phosphate cytidylyltransferase from Staphylococcus aureus: examination of structural basis for kinetic mechanism. AB - Integrity of the cell wall is essential for bacterial survival, and as a consequence components involved in its biosynthesis can potentially be exploited as targets for antibiotics. One such potential target is CTP:glycerol-3-phosphate cytidylyltransferase. This enzyme (TarD(Sa) in Staphylococcus aureus and TagD(Bs) in Bacillus subtilis) catalyzes the formation of CDP-glycerol, which is used for the assembly of linkages between peptidoglycan and teichoic acid polymer in Gram positive bacteria. Intriguingly, despite the high sequence identity between TarD(Sa) and TagD(Bs) (69% identity), kinetic studies show that these two enzymes differ markedly in their kinetic mechanism and activity. To examine the basis for the disparate enzymological properties, we have determined the crystal structure of TarD(Sa) in the apo state to 3 A resolution, and performed equilibrium sedimentation analysis. Comparison of the structure with that of CTP- and CDP glycerol-bound TagD(Bs) crystal structures reveals that the overall structure of TarD(Sa) is essentially the same as that of TagD(Bs), except in the C-terminus, where it forms a helix in TagD(Bs) but is disordered in the apo TarD(Sa) structure. In addition, TarD(Sa) can exist both as a tetramer and as a dimer, unlike TagD(Bs), which is a dimer. These observations shed light on the structural basis for the differing kinetic characteristics between TarD(Sa) and TagD(Bs). PMID- 16344012 TI - Synergism of Leu-Lys rich antimicrobial peptides and chloramphenicol against bacterial cells. AB - To investigate the antibiotic activity and synergistic effect, analogues were designed to increase not only net positive charge by Lys-substitution but also hydrophobic helix region by Leu-substitution from CA (1-8)-MA (1-12) hybrid peptide (CA-MA). In particular, CA-MA analogue P5 (P5), designed by flexible region (GIG-->P)-substitution, Lys- (positions 4, 8, 14, 15) and Leu- (positions 5, 6, 12, 13, 16, 17, 20) substitutions, showed potent antibacterial activity in minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) without having hemolytic activity. In addition, P5 and chloramphenicol has potent synergistic effect against tested cell lines. As determined by propidium iodide (PI) staining, flow cytometry showed that P5 plus chloramphenicol-treated cells had higher fluorescence intensity than untreated, P5- and chloramphenicol treated cells. The effect on plasma membrane was examined by investigating the transmembrane potential depolarizing experiments of S. aureus with P5 and chloramphenicol. The result showed that the peptide exerts its antibacterial activity by acting on the plasma membrane. Furthermore, P5 caused significant morphological alterations of S. aureus, as shown by scanning electron microscopy. Our results suggest that peptide P5 is an excellent candidate as a lead compound for the development of novel anti-infective agents and synergistic effects with conventional antibiotic agents but lack hemolytic activity. PMID- 16344013 TI - Trends in maternal health/healthcare in low-income countries and the implications on neonatal health. AB - Maternal health in low-income countries has received increasing attention over the last 15-20 years. Maternal mortality ratios in these countries are mainly still modelled estimates and one cannot discern trends. The introduction of registration systems-giving reliable causes of death-is essential both for monitoring maternal health and fuelling action. Countries with documented success in reducing maternal mortality have used systematic, incremental approaches, often tied to multi-sectoral efforts, including roads, communication links, education, water and sanitation. Improving maternal health requires a reasonably well-functioning health system and this typically requires the reduction of poverty and consistent investment in the health system. Increasingly, policies to improve maternal and newborn health are being implemented, but in the poorest countries the impact is still not discernible. New clinical interventions, such as effective treatment of (pre)eclampsia and post-partum haemorrhage, are often beyond reach of the majority in low-income countries. In summary, progress in maternal health, and the intimately linked perinatal and newborn health, is too slow and requires urgent efforts in poverty reduction and health system strengthening in low-income countries. PMID- 16344014 TI - Essential newborn care training activities: 8 years of experience in Eastern European, Caucasian and Central Asian countries. AB - There is still an alarming gap in neonatal healthcare and outcome between Western and Eastern European countries and the former USSR countries in particular. Most of the causes of neonatal mortality and morbidity can be prevented or managed by simple cost-effective interventions aimed at improving quality of healthcare, health system organisation and family and community participation. Training of health professionals and health policy-makers in the field of essential neonatal care and breastfeeding promotion is one of the cornerstones of the World Health Organization (WHO) initiatives Making Pregnancy Safer (MPS) and Promoting Effective Perinatal Care (PEPC) - the latter specifically tailored to the European Region - aimed at ensuring safe pregnancy and childbirth through ensuring the availability, access and use of quality skilled care. After 8 years of experience of training in essential neonatal care, positive changes in planning for and delivering neonatal care are taking place, even in challenging contexts, and this model of intervention should be further implemented in the region. PMID- 16344015 TI - Trends in perinatal care in developing countries. PMID- 16344016 TI - A new SNP in a negative regulatory region of the CYP19A1 gene is associated with lumbar spine BMD in postmenopausal women. AB - Osteoporosis is a common disease of bone possessing a strong genetic component. Cytochrome P450 aromatase, which is encoded by the CYP19A1 gene, converts androgens to estradiol. Considerable evidence suggests that extragonadal estrogens play an important role in determining bone mineral density (BMD) in postmenopausal women, and, among them, those synthesized in bone cells may also be important for the determination of bone phenotype. Therefore, CYP19A1 is an excellent candidate gene for osteoporosis. Since a region upstream of exon I.3, including exon I.6, was identified as containing repressor elements of promoter pII, we conducted a search for SNPs in this region of CYP19A1. Two SNPs [Aro1(rs4775936) and Aro2] located in exon I.6 and promoter I.6, respectively, were identified and their association with BMD analyzed in a cohort of 256 Spanish postmenopausal women. Aro1(rs4775936), but not Aro2, was associated with lumbar spine BMD (P = 0.029). Homozygotes AA (16% of the women) exhibited significantly higher lumbar spine BMD, compared with GG or GA individuals. Therefore, this study describes the Aro1 polymorphism which lies within a regulatory region and which may be a functional polymorphism, partially responsible for the bone phenotype it is associated with. PMID- 16344017 TI - Does standardized rehabilitation and discharge planning improve functional recovery in elderly patients with hip fracture? AB - OBJECTIVE: To determine whether standardized early rehabilitation and discharge planning increase risk-adjusted function and reduce risk-adjusted institutionalization in the first 6 months after hip fracture. DESIGN: Pre-post study of 2 independent population-based inception cohorts. SETTING: Two tertiary hospitals in an urban health region. PARTICIPANTS: Patients with hip fracture (N=919) 65 years and older. INTERVENTION: Subjects were enrolled before (control) and after (intervention) implementation of standardized rehabilitation and discharge planning. MAIN OUTCOME MEASURES: Function and institutionalization status were assessed at time of fracture and 3 and 6 months postfracture. Administrative databases provided length of stay (LOS) data. RESULTS: After risk adjustment, the Barthel Index score was significantly lower 3 months postfracture in control patients with low social support compared with those with higher social support (P<.05). Social support did not affect 3-month function in the intervention cohort. Control subjects with low social support were also significantly more likely to reside in long-term care by 6 months postfracture than intervention subjects with similar social support or those with higher social support (odds ratio=3.3; 95% confidence interval, 1.4-7.5). Total LOS did not change between cohorts. CONCLUSIONS: Overall, standardized rehabilitation and discharge planning did not affect postoperative function or institutionalization in elderly patients with hip fracture. In intervention patients with low social support, function improved and institutionalization was reduced. PMID- 16344018 TI - Venous thromboembolism after spinal cord injury: incidence, time course, and associated risk factors in 16,240 adults and children. AB - OBJECTIVE: To analyze the incidence of venous thromboembolism (VTE) after spinal cord injury (SCI). DESIGN: Retrospective cohort analysis of all SCI cases (16,240) in California from 1991 through 2001. SETTING: All public hospitals in California. PARTICIPANTS: Subjects (cases) coded as having complete or incomplete SCI. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Diagnosis of VTE or death within 91 days of the day of hospital admission. RESULTS: For all cases, the 91-day cumulative incidence of VTE was 5.4%. In a multivariate model, significant predictors of VTE included male sex (odds ratio [OR]=1.4; 95% confidence interval [CI], 1.2-1.7), African-American race (OR=1.6; 95% CI, 1.3 1.9), complete paraplegia versus tetraplegia (OR=1.8; 95% CI, 1.4-2.3), and presence of 3 or more comorbid conditions versus none (OR=1.6; 95% CI, 1.3-2.1). Age less than 14 years was predictive of not developing VTE (OR=0.2; 95% CI, 0.1 0.7). The incidence of VTE did not change significantly over the 11-year time period (P=.07), and VTE was not a significant predictor of death in the first 91 days after hospitalization. CONCLUSIONS: The incidence of VTE in SCI patients in California did not change between 1991 and 2001. We identified specific risk factors for VTE. Further studies are needed to determine if prompt initiation of medical prophylaxis in high risk subjects reduces the incidence of symptomatic VTE. PMID- 16344019 TI - Acupuncture for upper-extremity rehabilitation in chronic stroke: a randomized sham-controlled study. AB - OBJECTIVE: To compare the effects of traditional Chinese acupuncture with sham acupuncture on upper-extremity (UE) function and quality of life (QOL) in patients with chronic hemiparesis from stroke. DESIGN: A prospective, sham controlled, randomized controlled trial (RCT). SETTING: Patients recruited through a hospital stroke rehabilitation program. PARTICIPANTS: Thirty-three subjects who incurred a stroke 0.8 to 24 years previously and had moderate to severe UE functional impairment. INTERVENTIONS: Active acupuncture tailored to traditional Chinese medicine diagnoses, including electroacupuncture, or sham acupuncture. Up to 20 treatment sessions (mean, 16.9) over a mean of 10.5 weeks. MAIN OUTCOME MEASURES: UE motor function, spasticity, grip strength, range of motion (ROM), activities of daily living, QOL, and mood. All outcomes were measured at baseline and after treatment. RESULTS: Intention-to-treat (ITT) analyses found no statistically significant differences in outcomes between active and sham acupuncture groups. Analyses of protocol-compliant subjects revealed significant improvement in wrist spasticity (P<.01) and both wrist (P<.01) and shoulder (P<.01) ROM in the active acupuncture group, and improvement trends in UE motor function (P=.09) and digit ROM (P=.06). CONCLUSIONS: Based on ITT analyses, we conclude that acupuncture does not improve UE function or QOL in patients with chronic stroke symptoms. However, gains in UE function observed in protocol-compliant subjects suggest traditional Chinese acupuncture may help patients with chronic stroke symptoms. These results must be interpreted cautiously because of small sample sizes and multiple, unadjusted, post hoc comparisons. A larger, more definitive RCT using a similar design is feasible and warranted. PMID- 16344020 TI - Geriatric depression, medical diagnosis, and functional recovery during acute rehabilitation. AB - OBJECTIVE: To examine and compare the prevalence and functional impact of depressive symptoms for older adult stroke and nonstroke rehabilitation inpatients. DESIGN: Case-control study examining functional outcome using a 2 (stroke, nonstroke) by 2 (depression, no depression) design. SETTING: Urban hospital rehabilitation unit. PARTICIPANTS: A total of 509 rehabilitation inpatients (age, > or = 60 y) were included and grouped by diagnosis of stroke (n=207) and nonstroke (n=302). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Geriatric Depression Scale and FIM instrument. Analysis of covariance procedures examined the impact of depressive symptoms on discharge functional ability controlling for age, sex, admission functional ability, and hospital length of stay. RESULTS: Prevalence of depressive symptoms was similar for stroke (31.8%) and nonstroke (31.5%) and negatively associated with functional ability at discharge for both groups. Overall, the stroke and nonstroke groups did not differ significantly with respect to functional recovery. CONCLUSIONS: Depression, and its impact on acute rehabilitation, is significantly related to functional recovery but does not differ in its frequency or impact for stroke patients. Because depressive symptoms do not appear to discriminate across diagnostic groups, routine screening for depression is recommended for all rehabilitation inpatients. PMID- 16344021 TI - Prehabilitation and rehabilitation for attenuating hindlimb unweighting effects on skeletal muscle and gait in adult and old rats. AB - OBJECTIVE: To compare the effectiveness of no exercise with prehabilitation (exercise before hindlimb unweighting [HLU]) versus rehabilitation (exercise given after HLU) on gait function and skeletal muscle mass and force. DESIGN: Randomized controlled trial. SETTING: Animal laboratory. ANIMALS: Male-specific, pathogen-free Fisher344/Brown Norway rats (N=149). Groups consisted of adult and old controls, HLU, prehabilitation, rehabilitation, natural cage recovery (reloading), and exercise without HLU. INTERVENTIONS: Ten days of general conditioning exercise were given to 6-month-old adult and 30-month-old old rats before or after a week of HLU. MAIN OUTCOME MEASURES: Gait stride length and width; soleus, plantaris, extensor digitorum longus, and peroneus longus mass and peak contractile force; whole gastrocnemius mass; and total protein concentration for the soleus and gastrocnemius. RESULTS: Muscle mass (approximately 30%) and force (24%-36%) declined with age in all muscles studied. In adult rats declines in muscle mass occurred with HLU in the soleus, plantaris, and gastrocnemius. Prehabilitation did not prevent the loss of muscle mass in adult rats. Rehabilitation and natural recovery effectively restored soleus and gastrocnemius muscle mass in adult rats but not soleus peak force. Old rats had a significant 23% HLU effect only on gastrocnemius mass (control, 1670+/-129 mg; HLU, 1274+/ 184 mg). Prehabilitation did not prevent the decline in gastrocnemius mass. Rehabilitation in old rats restored gastrocnemius mass to within 13% of control levels. Prehabilitation was effective for preventing and rehabilitation was effective for restoring soleus contractile force in old rats (control, 114+/-9 mg; HLU, 67+/-22 mg; prehabilitation, 106+/-31 mg; rehabilitation, 120+/-26 mg) compared with recovery without exercise (86+/-29 g). A significant reduction in stride length was observed with aging (136+/-18 mm vs 98+/-10 mm), which decreased further with HLU (78+/-14 mm). Prehabilitation attenuated HLU-related reductions in stride length, and rehabilitation was effective for stride length restoration in old rats. CONCLUSIONS: Exercise, particularly rehabilitation, was more effective for old than young rats. Prehabilitation and rehabilitation diminished some of the detrimental effects of HLU on skeletal muscle mass and force and gait function in old rats. PMID- 16344022 TI - The validity of prospective and retrospective global change criterion measures. AB - OBJECTIVE: To assess the validity of retrospective versus prospective criterions of change. DESIGN: Single cohort pretest-posttest design. SETTING: Physical or occupational therapy outpatient clinics. PARTICIPANTS: Volunteer sample of 211 patients with upper-extremity musculoskeletal problems. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Disabilities of the Arm, Shoulder, and Hand questionnaire, the Shoulder Pain and Disability Index, the Patient-Rated Wrist Evaluation, the Medical Outcomes Study 12-Item Short-Form Health Survey; global disability rating (GDR), retrospective global rating of change (GRC), and patient satisfaction. RESULTS: Correlations were calculated among the baseline, 3-month follow-up, and change scores for each outcome measure with the change criterion instruments. Retrospective GRC and patient satisfaction ratings showed moderate correlations with the 3-month follow-up scores, but nonsignificant correlations with baseline scores. By contrast, the prospective GDR criterion showed significant correlations with both baseline and 3-month follow-up scores ranging between 0.3 and 0.4 (absolute value). CONCLUSIONS: Retrospective self-report measures of change do not accurately reflect true change over time. The retrospective GRC and patient satisfaction were heavily influenced by current (posttreatment) status whereas the prospective global change measure reflected both baseline and posttreatment status equally and thus appeared to be a more valid measure of change over time. This study demonstrates the need for an alternative criterion for establishing true individual change. PMID- 16344023 TI - Measuring activity limitations in walking: development of a hierarchical scale for patients with lower-extremity disorders who live at home. AB - OBJECTIVE: To develop a hierarchical scale that measures activity limitations in walking in patients with lower-extremity disorders who live at home. DESIGN: Cross-sectional study. SETTING: Orthopedic workshops and outpatient clinics of secondary and tertiary care centers. PARTICIPANTS: Patients (N=981; mean age +/- standard deviation, 58.6+/-15.4 y; 46% men) living at home, with different lower extremity disorders: stroke, poliomyelitis, osteoarthritis, amputation, complex regional pain syndrome type I, and diabetic and degenerative foot disorders. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: (1) Fit of the monotone homogeneity model, indicating whether items can be used for measuring patients; (2) fit of the double monotonicity model, indicating invariant (hierarchical) item ordering; (3) intratest reliability, indicating repeatability of the sum score; (4) robustness, addressing the clinimetric properties within subgroups of patients; and (5) differential item functioning, addressing the validity of comparisons between subgroups of patients. RESULTS: Thirty-five of 41 dichotomous items had (1) good fit of the monotone homogeneity model (coefficient H=.50), (2) good fit of the double monotonicity model (coefficient H(T)=.33), (3) good intratest reliability (coefficient rho=.95), (4) satisfactory robustness (within subgroups of patients defined by age, sex, and diagnosis), and (5) some differential item functioning (6 items in amputees compared with nonamputees). CONCLUSIONS: A hierarchical scale, with excellent scaling characteristics, was developed to measure activity limitations in walking in patients with lower extremity disorders who live at home. The measurements should be interpreted cautiously when making comparisons between amputees and nonamputees. PMID- 16344024 TI - Efficacy and safety of oxymorphone immediate release for the treatment of mild to moderate pain after ambulatory orthopedic surgery: results of a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To assess the analgesic efficacy and safety of 5 mg of oxymorphone immediate release (IR) for mild to moderate pain. DESIGN: Multicenter, double blind, randomized, placebo-controlled study. SETTING: Ambulatory surgical centers. PARTICIPANTS: Outpatients (age, > or = 18 y) undergoing knee arthroscopy. INTERVENTION: Randomization to 5 mg of oxymorphone IR or placebo hourly as needed for up to 8 hours. MAIN OUTCOME MEASURE: Sum of pain intensity difference (SPID) from baseline to 8 hours. RESULTS: Among 122 patients randomized, 70.5% and 28.7% had moderate or mild postsurgical pain at baseline, respectively. The mean SPID score was significantly greater in the oxymorphone IR group, showing greater pain relief, compared with the placebo group (least squares mean difference +/- standard error, 76.9+/-28.09; 95% confidence interval, 21.26-132.59; P=.007). More placebo patients (48.4%) required rescue medication than oxymorphone IR patients (16.7%), with median times to use of rescue medication of 6 hours 54 minutes and more than 8 hours, respectively (P<.001). More patients (47.4%) rated oxymorphone IR "very good" or "excellent" for pain relief versus placebo (25.0%). No oxymorphone IR-treated patients discontinued because of adverse events (AEs) or experienced serious AEs. CONCLUSIONS: Five milligrams of oxymorphone IR was well tolerated and effective at relieving mild or moderate postsurgical pain after outpatient knee surgery. PMID- 16344025 TI - Health status rated with the Medical Outcomes Study 36-Item Short-Form Health Survey after spinal cord injury. AB - OBJECTIVE: To describe the health status of Australians with spinal cord injury (SCI). DESIGN: Survey. SETTING: Australian population-based sample. PARTICIPANTS: Participants (N=305) with SCI at recruitment to a randomized trial. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Medical Outcomes Study 36 Item Short-Form Health Survey (SF-36) health questionnaire. RESULTS: Compared with the general population, our sample reported significantly lower scores in 6 SF-36 domains (physical function, role-physical, bodily pain, general health, social function, vitality) and the physical component summary (PCS) score, but unexpectedly higher mental component summary (MCS) scores (difference in mean MCS scores, 4.6; 95% confidence interval [CI], 2.4-6.8). Compared with people with tetraplegia, those with paraplegia had better physical function and PCS scores (difference, 21; 95% CI, 17-24; difference, 3; 95% CI, 1-5, respectively), and worse bodily pain scores (difference, 9; 95% CI, 2-15). Recent urinary infections were associated with worse general health, vitality, mental health, and MCS scores. Receiving family or external physical care was associated with worse physical function and PCS scores, but better mental health and MCS scores. Older age at injury was associated with lower bodily pain and PCS scores. CONCLUSIONS: Using the SF-36, Australians with SCI rate their physical (not mental) health status as worse than the general population. PMID- 16344027 TI - The relation between subjective and objective measures of everyday life activities in persons with multiple sclerosis. AB - OBJECTIVES: To investigate the relation between subjective and objective performance-based measures of functional status in persons with multiple sclerosis (MS), and to compare their performance with healthy controls. DESIGN: A between-groups design, using a correlational approach to examine the relation between objective and subjective measures of functional capacity. SETTING: Outpatient rehabilitation research institution. PARTICIPANTS: Seventy-four subjects with clinically definite MS and 35 healthy controls. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The Executive Function Performance Test (EFPT), Functional Assessment of Multiple Sclerosis (FAMS), and Functional Behavior Profile (FBP). RESULTS: MS participants reported more difficulties performing functional tasks than did the healthy controls. MS participants also performed significantly worse on the EFPT than healthy controls. However, all correlations between subjective and objective functional measures were nonsignificant. After controlling for depressive symptomatology, EFPT performance was significantly associated with FBP scores, but not FAMS scores. CONCLUSIONS: The lack of association between objective performance-based measures and subjective self-report measures of functional activities is a challenge to outcomes measurement and has implications for assessment of functional performance. Results are discussed in terms of the different dimensions that these tools are measuring and their respective strengths and limitations. PMID- 16344026 TI - Outcome of a comprehensive neurorehabilitation program for patients with traumatic brain injury. AB - OBJECTIVE: To evaluate the outcome of a comprehensive neurorehabilitation program compared with that of conventional clinical care and rehabilitation for patients with traumatic brain injury (TBI). DESIGN: Nonrandomized, controlled trial with a 2-year follow-up. SETTING: Nationwide rehabilitation center and level I trauma center, both in Finland. PARTICIPANTS: We studied 19 consecutive adults with a significant TBI who underwent a comprehensive neurorehabilitation program and 20 control patients who received conventional rehabilitation referred by physicians in the general health care system. The outcome of the control patients was not known before the selection. The groups were similar in age, sex, education, injury severity (assessed on the Glasgow Coma Scale, radiologic and neuropsychologic findings, neurosurgical interventions), time from the injury, and preinjury employment status. INTERVENTIONS: A postacute, intensive, interdisciplinary, 6-week rehabilitation program for TBI patients who are considered to have adequate potential to achieve productivity by this means; focus on neuropsychologic rehabilitation and psychotherapy with vocational interventions and follow-up support. MAIN OUTCOME MEASURE: Status of productivity, judged as productive (defined as working, studying, or participating in volunteer activities) or nonproductive, evaluated on questionnaires filled in by patients and their significant others at the time of follow-up evaluation. RESULTS: At follow-up, 89% of the treated patients were productive compared with 55% of the controls. The rehabilitation program was significantly predictive of the productive status at follow-up (odds ratio=6.96; 95% confidence interval, 1.26-38.44; P=.017). Other factors did not explain the better productivity of the treatment group. Two neuropsychologist-evaluators, who were blind to the rehabilitation history of patients and to each other's evaluations, were perfectly consistent in their classification of patients' productivity statuses. CONCLUSIONS: The findings support the proposition that comprehensive neuropsychologically oriented rehabilitation programs can improve psychosocial functioning in terms of productivity in postacute patients with moderate to severe TBI. Additional larger controlled studies are needed to establish the efficacy of TBI rehabilitation interventions. PMID- 16344028 TI - Postural control of the lumbar spine in unstable sitting. AB - OBJECTIVE: To evaluate the neuromuscular strategy adopted during sitting balance on an unstable surface in the frontal plane. DESIGN: Electromyographic evaluation of trunk muscles. SETTING: University spine biomechanics laboratory. PARTICIPANTS: Seventy asymptomatic men (mean age, 34.5 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: "Balancers" and "nonbalancers" were identified by principal component analysis of their lumbar spine side flexion angle during sitting balance. Average electromyographic levels were used as a measure of muscle activation. Pearson correlations were used to identify coactivation versus asymmetrical muscle activation of opposite muscle groups. RESULTS: External oblique, internal oblique, and thoracic erector spinae (TES) were most active, and most likely to be used asymmetrically, with other muscles showing low levels of coactivation. Between groups, the average electromyographic levels in the balancers was lower than in the nonbalancers (P<.05), with further differences in the symmetry of external oblique, internal oblique, and TES activation between groups. CONCLUSIONS: Sitting balance in the frontal plane appears to involve a combined feedforward-feedback strategy of muscle activation. Successful balance was characterized by low levels of muscle coactivity, along with higher levels of asymmetric activation in the global trunk muscles, specifically external oblique, internal oblique, and TES. PMID- 16344029 TI - Wheelchair skills training for community-based manual wheelchair users: a randomized controlled trial. AB - OBJECTIVE: To test the hypotheses that wheelchair skills training of community based manual wheelchair users is efficacious, safe, and practical. DESIGN: Randomized controlled trial. SETTING: Rehabilitation center and community. PARTICIPANTS: Twenty community-based manual wheelchair users (15 men, 5 women; age range, 21-77 y), half with musculoskeletal and half with neurologic disorders. INTERVENTION: Participants were randomly allocated to the Wheelchair Skills Training Program (WSTP) or control groups. In 1-hour individualized sessions, the WSTP group participants received a mean +/- standard deviation of 4.5+/-0.7 hours of training. Caregivers participated whenever possible. In addition to training at the rehabilitation center, the trainer traveled to administer training in the community. MAIN OUTCOME MEASURES: Using the Wheelchair Skills Test (WST, version 3.1), an objective test of 57 skills, we calculated total and subtotal percentage scores (percentage number of skills passed of those possible) and individual skill success rates. RESULTS: The WSTP group's improvement in total WST score was significantly greater than the control group's (P<.005). The mean total WST score for the WSTP group increased from a pretraining value of 63.3%+/-6.0% to 78.5%+/-8.3% posttraining, a relative improvement of 24.0% (P=.002). The control group increased from a baseline value of 70.8%+/-14.0% to 74.2%+/-11.8% at follow-up, a relative improvement of 4.8% (P=.03). The WSTP group had clinically significant pre- and posttraining improvements (> or = 20%) in the success rates of 25 of the 57 individual WST skills, compared with only 5 skills for the control group. There were no adverse incidents, and the WSTP participants' comments were all positive. CONCLUSIONS: Wheelchair skills training of community-based manual wheelchair users is efficacious, safe, and practical. These findings have implications for the standard of rehabilitation care. PMID- 16344030 TI - Comparative effectiveness of the GivMohr sling in subjects with flaccid upper limbs on subluxation through radiologic analysis. AB - OBJECTIVE: To test the effectiveness of the GivMohr sling in reducing subluxation while providing joint compression through a flaccid limb, using the criterion standard of radiography for measuring subluxation. DESIGN: Anteroposterior 0 degree radiographs were taken of each subject: 1 of the unaffected shoulder and 3 of the affected shoulder; 1 without a sling, 1 with the GivMohr sling, and 1 with the Rolyan humeral cuff sling. SETTING: Two large rehabilitation centers. PARTICIPANTS: Twenty-five adult volunteers with a flaccid upper limb (UL) secondary to cerebrovascular accident or other pathology. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Vertical and horizontal subluxation were measured in millimeters on each of the 4 radiographs for each subject. RESULTS: A 1-way analysis of variance revealed a significant main effect for vertical but not horizontal subluxation. Post hoc tests showed that the GivMohr sling measures were similar to measures for the uninvolved shoulder, but significantly different from measures for the Rolyan and the involved shoulder. CONCLUSIONS: These results show that a properly fitted GivMohr sling reduces subluxation without overcorrecting. This new sling provides an alternative treatment option for persons with flaccid ULs that may prevent secondary complications and improve outcomes. PMID- 16344031 TI - Feedback-controlled and programmed stretching of the ankle plantarflexors and dorsiflexors in stroke: effects of a 4-week intervention program. AB - OBJECTIVE: To investigate the effect of repeated feedback-controlled and programmed "intelligent" stretching of the ankle plantar- and dorsiflexors to treat subjects with ankle spasticity and/or contracture in stroke. DESIGN: Noncontrolled trial. SETTING: Institutional research center. PARTICIPANTS: Subjects with spasticity and/or contracture after stroke. INTERVENTIONS: Stretching of the plantar- and dorsiflexors of the ankle 3 times a week for 45 minutes during a 4-week period by using a feedback-controlled and programmed stretching device. MAIN OUTCOME MEASURES: Passive and active range of motion (ROM), muscle strength, joint stiffness, joint viscous damping, reflex excitability, comfortable walking speed, and subjective experiences of the subjects. RESULTS: Significant improvements were found in the passive ROM, maximum voluntary contraction, ankle stiffness, and comfortable walking speed. The visual analog scales indicated very positive subjective evaluation in terms of the comfort of stretching and the effect on their involved ankle. CONCLUSIONS: Repeated feedback-controlled or intelligent stretching had a positive influence on the joint properties of the ankle with spasticity and/or contracture after stroke. The stretching device may be an effective and safe alternative to manual passive motion treatment by a therapist and has potential to be used to repeatedly and regularly stretch the ankle of subjects with spasticity and/or contracture without daily involvement of clinicians or physical therapists. PMID- 16344032 TI - The reliability and validity of assessment of sagittal plane deviations in children who have spastic diplegia. AB - OBJECTIVE: To assess the reliability and validity of a newly described classification of sagittal plane alignment in spastic diplegic gait. DESIGN: Twenty split-screen videos of children with spastic diplegia, Gross Motor Function Classification System levels I to III, were viewed on 2 occasions, 6 weeks apart, by 5 raters. The sagittal plane alignments of the right and left lower limbs in gait were classified separately as true equinus, jump knee, apparent equinus, or crouch, based on the published classification. A fifth category, nonclassifiable, was used if classification was not possible. We then used sagittal plane kinematic data to confirm the classification for each subject and these were compared with rater classification scores, which used the video information only. SETTING: Tertiary-level children's hospital. PARTICIPANTS: Three pediatric orthopedic surgeons and 2 pediatric orthopedic residents. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Gait classification scores derived from visual observation were compared among and within raters. The gait classification scores derived from visual observation were compared with the scores derived from sagittal plane kinematic data to assess validity. RESULTS: A moderate correlation was found among the 5 raters within each session, with an interrater weighted kappa score of .45 in session 1 and .49 in session 2. The intrarater, weighted kappa scores showed a moderate to substantial level of agreement between sessions, ranging from .50 to .68. The classification scores of individual raters had moderate validity when compared with classifications derived from the sagittal plane kinematic data. However, there was a substantial level of agreement between the consensus opinion and the classification obtained using the kinematic data as well as the video recordings (weighted kappa=0.8). CONCLUSIONS: This classification has only moderate reliability and validity when a single experienced rater views the 2-dimensional gait videos. However, the consensus opinion derived from the scores of 5 raters considerably improves the validity of the assessment. PMID- 16344033 TI - Repeatability of temporospatial gait measures in children using the GAITRite electronic walkway. AB - OBJECTIVE: To determine the repeatability of selected temporal and spatial gait parameters in healthy children using the GAITRite electronic walkway. DESIGN: Immediate test-retest design. SETTING: Community. PARTICIPANTS: A sample of 57 subjects (age range, 1.3-10.9 y) participated. Subjects were stratified into 3 age groups (1.0 to <4.0 years old, 4.0 to <8.0 years old, 8.0 to <11.0 years old). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Subjects completed 2 barefooted test walks at preferred gait speed. Intraclass correlation coefficient (ICC(1,1)) values, 95% confidence intervals, the Bland-Altman limits of agreement, and coefficient of variation (CV) were calculated. RESULTS: Overall, ICC values ranged from .05 to .93. The amount of clinically relevant error (CV) was acceptable for all gait variables across age groups with the exception of the toe-in/out variables (CV, 46.7%-92.9%) and the heel-to-heel base of support variable (CV, 18.6%-31.5%). CONCLUSIONS: Repeatability of GAITRite measurements to assess temporospatial gait parameters in typically developing children walking at self-selected paces ranged from poor to excellent. Our repeatability data for child subjects between the ages of 1.3 to 10.9 years are comparable to adult data for the same temporospatial gait parameters. We believe that the GAITRite electronic walkway is an emerging clinical tool for assessment of gait in children with and without disabilities. PMID- 16344034 TI - Inter- and intrarater reliability of a back range of motion instrument. AB - OBJECTIVE: To examine the interrater and intrarater reliability of a back range of motion (BROM) instrument when measuring lumbar spine active planar motions and pelvic inclination. DESIGN: Single-group repeated measures for inter- and intrarater reliability. SETTING: Academic institution. PARTICIPANTS: Ninety-one participants (61 women, 30 men; mean age, 28 y) without a current complaint of low back pain volunteered. INTERVENTION: Two examiners measured pelvic inclination and all lumbar motions by using the BROM device. Subjects alternated between examiners for 4 complete trials; examiners remained blinded to the measurements. MAIN OUTCOME MEASURES: Intraclass correlation coefficients (ICCs) were used to determine intrarater and interrater reliability. Regression analysis was performed to determine the role palpation played in sagittal plane measurement error. RESULTS: Intrarater reliability for side bending was good (ICC range, .85-.83), lumbar forward flexion and pelvic inclination was good to fair (ICC range, .84-.79), and extension and rotation was fair to poor (ICC range, .76 .58). Interrater reliability was fair to poor for all lumbar motions and for pelvic inclination (ICC range, .79-.55). Less than 2% of the variation in sagittal plane measurements was explained by consistency of palpation for device placement. CONCLUSIONS: The BROM provides a reliable means of measuring lumbar forward flexion, side bending, and pelvic inclination when performed by the same examiner in asymptomatic subjects. PMID- 16344035 TI - Intra- and interrater reliability of the Ergo-Kit functional capacity evaluation method in adults without musculoskeletal complaints. AB - OBJECTIVE: To evaluate the intra- and interrater reliability of tests from the Ergo-Kit (EK) functional capacity evaluation method in adults without musculoskeletal complaints. DESIGN: Within-subjects design. SETTING: Academic medical center in The Netherlands. PARTICIPANTS: Twenty-seven subjects without musculoskeletal complaints (15 men, 12 women). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Seven EK tests (2 isometric, 3 dynamic lifting, 2 manipulation tests) were each assessed 3 times (over 4 days), twice by 1 rater (R1) and once by another rater (R2). Intrarater reliability was calculated using the EK test scores assessed by R1. Interrater reliability was calculated using the EK test scores assessed by both raters. Counterbalancing the rater order made possible the calculation of 2 interrater reliability levels (at time intervals of 4 and 8d). All reliability levels were expressed as intraclass correlation coefficients (ICCs). RESULTS: Intrarater and interrater reliability (8-d time interval) was high (ICC, >.80) for the isometric lifting tests, moderate (ICC range, .50-.80) for the dynamic lifting tests, and low (ICC, <.50) for the manipulation tests. The interrater reliability of the isometric and dynamic lifting tests (4-d time interval) was high (ICC, >.80), and it was moderate (ICC range, .50-.80) for both manipulation tests. CONCLUSIONS: The isometric and dynamic lifting tests of the EK have a moderate to high level of reliability; the manipulation tests have a low level of reliability. PMID- 16344036 TI - Evaluation of the safety and durability of low-cost nonprogrammable electric powered wheelchairs. AB - OBJECTIVE: To evaluate whether a selection of low-cost, nonprogrammable electric powered wheelchairs (EPWs) meets the American National Standards Institute (ANSI)/Rehabilitation Engineering and Assistive Technology Society of North America (RESNA) Wheelchair Standards requirements. DESIGN: Objective comparison tests of various aspects of power wheelchair design and performance of 4 EPW types. SPECIMENS: Three of each of the following EPWs: Pride Mobility Jet 10 (Pride), Invacare Pronto M50 (Invacare), Electric Mobility Rascal 250PC (Electric Mobility), and the Golden Technologies Alante GP-201-F (Golden). SETTING: Rehabilitation engineering research center. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Static tipping angle; dynamic tipping score; braking distance; energy consumption; climatic conditioning; power and control systems integrity and safety; and static, impact, and fatigue life (equivalent cycles). RESULTS: Static tipping angle and dynamic tipping score were significantly different across manufacturers for each tipping direction (range, 6.6 degrees-35.6 degrees). Braking distances were significantly different across manufacturers (range, 7.4-117.3 cm). Significant differences among groups were found with analysis of variance (ANOVA). Energy consumption results show that all EPWs can travel over 17 km before the battery is expected to be exhausted under idealized conditions (range, 18.2-32.0 km). Significant differences among groups were found with ANOVA. All EPWs passed the climatic conditioning tests. Several adverse responses were found during the power and control systems testing, including motors smoking during the stalling condition (Electric Mobility), charger safety issues (Electric Mobility, Invacare), and controller failures (Golden). All EPWs passed static and impact testing; 9 of 12 failed fatigue testing (3 Invacare, 3 Golden, 1 Electric Mobility, 2 Pride). Equivalent cycles did not differ statistically across manufacturers (range, 9759-824,628 cycles). CONCLUSIONS: Large variability in the results, especially with respect to static tipping, power and control system failures, and fatigue life suggest design improvements must be made to make these low-cost, nonprogrammable EPWs safe and reliable for the consumer. Based on our results, these EPWs do not, in general, meet the ANSI/RESNA Wheelchair Standards requirements. PMID- 16344037 TI - Test methods to detect hip and knee muscle weakness and gait disturbance in patients with hip osteoarthritis. AB - OBJECTIVE: To evaluate test methods for hip and knee muscle weakness and gait disturbance. DESIGN: Test-retest. SETTING: Orthopedic university clinic. PARTICIPANTS: Ten young (age, 36+/-6 y) and 13 elderly (age, 69+/-8 y) healthy volunteers and 11 patients (age, 69+/-8 y) with unilateral hip osteoarthritis (OA) were tested for muscular strength. Twenty-five volunteers (age, 42+/-14 y) underwent gait analysis. INTERVENTIONS: A dynamometer assessing maximal voluntary isometric force of hip and knee muscles and an optosensor walkway detecting limp were developed. Tests evaluated reproducibility and tolerance in patients with OA and elderly subjects. MAIN OUTCOME MEASURES: Relative coefficient of variation (CV%) and force (in newtons). RESULTS: CV% for unilateral strength measurements ranged from 7% to 12% for specific muscle groups. CV% for gait parameters ranged from 4% to 8%, except for the double-support phase. Tests were well tolerated, and no patient had to discontinue because of fatigue. Differences related to sex, age, and disease were detected. CONCLUSIONS: Our dynamometer system provides reliable measurements of hip and knee muscle strength in young and old people, and variation is comparable to previous data. Our photocell technique for gait analysis is reliable in people with normal gait. Both methods are attractive because they are affordable, nonstationary, and easy to use. PMID- 16344038 TI - Management of diabetes during acute stroke and inpatient stroke rehabilitation. AB - OBJECTIVES: To summarize evidence on the impact of hyperglycemia on stroke outcomes and to present therapy algorithms for inpatient management in diabetic stroke patients. DATA SOURCES: Guidelines for inpatient management of diabetes were reviewed and extracted from a technical review and recommendations from 2 national diabetes and endocrine organizations. MEDLINE database searches were conducted using key words: stroke, diabetes, hyperglycemia, hypoglycemia, inpatient, hospitalized, treatment, outcomes, disability, self-management, and education. STUDY SELECTION: Studies were selected that specifically addressed the impact of the following in stroke patients: hyperglycemia and diabetes on rehabilitation outcomes, management strategies for hyperglycemia and diabetes, and strategies for facilitating diabetes self-management. DATA EXTRACTION: Two authors independently extracted data and management practices from selected articles and published practice guidelines. DATA SYNTHESIS: Diabetes is prevalent in stroke patients and results in poorer inpatient hospital and rehabilitation outcomes. Management of diabetes in stroke patients is further complicated by impairments in mobility and vision, necessitating accommodation strategies and tools for self-management. Optimal management of hyperglycemia using insulin or oral hypoglycemic agents results in reduced morbidity and mortality among diabetic inpatients. CONCLUSIONS: To achieve inpatient glycemic management targets, use of clinical management algorithms, self-management tools, and systems approaches such as diabetes management teams are useful. PMID- 16344039 TI - "Ineffectiveness" of automated locomotor training. PMID- 16344041 TI - Control of cytokine gene expression by PG101, a water-soluble extract prepared from Lentinus lepideus. AB - A water-soluble extract from Lentinus lepideus mycelium, named PG101, has been shown to control the expression of various cytokines [M. Jin, H.J. Jung, J.J. Choi, H.Jeon, J.H. Oh, B. Kim, S.S. Shin, J.K. Lee, K. Yoon, S. Kim, Activation of selective transcription factors and cytokines by water-soluble extract from Lentinus lepideus, Exp. Biol. Med. (Maywood) 228 (2003) 749-758]. To understand its molecular mechanism(s), PG101-mediated activation of cytokines was studied at the RNA and protein levels. Results from Northern blot analysis indicated that the steady-state RNA levels of TNF-alpha and seven other cytokines were highly increased in human peripheral blood mononuclear cells treated with PG101. The RNA level of TNF-alpha, MIP-1alpha, MIP-1beta, MIP-3alpha, and IL-8 was not affected by the presence of cycloheximide, an inhibitor of the translation process, suggesting that they are the direct targets of PG101. A significantly high protein level of TNF-alpha, MIP-1alpha, and IL-8 remained detectable, even when cells were cultured with actinomycin D, 2h prior to the PG101 treatment. Our data indicate that PG101 controls selective cellular proteins, which play key roles in the innate immune system, at the transcriptional and post-translational levels. PMID- 16344044 TI - Helicobacter pylori infection and surgical disease--part II. PMID- 16344047 TI - Barry Marshall 2005 Nobel laureate in medicine and physiology. PMID- 16344048 TI - Have we oversold colonoscopy? PMID- 16344049 TI - Glucagonoma presenting with necrolytic migratory erythema: the Glucagonoma syndrome. PMID- 16344050 TI - Neural stem cell transplantation in the stomach rescues gastric function in neuronal nitric oxide synthase-deficient mice. AB - BACKGROUND & AIMS: Nitric oxide is a major inhibitory neurotransmitter in the enteric nervous system. Loss or dysfunction of nitrinergic neurons is associated with serious disruptions of motility, intractable symptoms, and long-term suffering. The aim of this study was to evaluate the effect of intrapyloric transplantation of neural stem cells (NSCs) on gastric emptying and pyloric function in nNOS-/- mice, a well-established genetic model of gastroparesis. METHODS: NSCs were isolated from embryonic mice transgenically engineered to express green fluorescent protein and transplanted into the pylorus of nNOS-/- mice. Grafted cells were visualized in pyloric sections and further characterized by immunofluorescence staining. One week posttransplantation, gastric emptying to a non-nutrient meal was measured using the phenol red method and pyloric function was assessed by measuring the relaxation of pyloric strips in an organ bath in response to electrical field stimulation (EFS) under nonadrenergic, noncholinergic conditions. RESULTS: One week following implantation, grafted NSCs differentiated into neurons and expressed neuronal nitric oxide synthase. Gastric emptying was significantly increased in mice that received NSCs as compared with vehicle-injected controls (49.67% vs 35.09%; P < .01 by Student t test). EFS induced relaxation of pyloric strips was also significantly increased (P < .01 by 2-way analysis of variance). The nitric oxide synthase inhibitor N(G)-nitro-l arginine methyl ester and the neuronal blocker tetrodotoxin blocked the EFS induced relaxation, indicating that the observed effect is NO mediated and neuronally derived. CONCLUSIONS: Our results support the potential of NSC transplantation as a viable therapeutic option for neuroenteric disorders. PMID- 16344051 TI - Prevalence of Barrett's esophagus in the general population: an endoscopic study. AB - BACKGROUND & AIMS: Barrett's esophagus (BE) is associated with esophageal adenocarcinoma, the incidence of which has been increasing dramatically. The prevalence of BE in the general population is uncertain because upper endoscopy is required for diagnosis. This study aimed to determine the prevalence of BE and possible associated risk factors in an adult Swedish population. METHODS: A random sample (n = 3000) of the adult population (n = 21,610) in 2 municipalities was surveyed using a validated gastrointestinal symptom questionnaire (response rate, 74%); a random subsample (n = 1000; mean age, 53.5 years; 51% female) underwent upper endoscopy. Endoscopic signs suggestive of columnar-lined esophagus (CLE) were defined as mucosal tongues or an upward shift of the squamocolumnar junction. BE was diagnosed when specialized intestinal metaplasia was detected histologically in suspected CLE. RESULTS: BE was present in 16 subjects (1.6%; 95% confidence interval, 0.8-2.4): 5 with a long segment and 11 with a short segment. Overall, 40% reported reflux symptoms and 15.5% showed esophagitis; 103 (10%) had suspected CLE, and 12 (1.2%) had a visible segment > or = 2 cm. The prevalence of BE in those with reflux symptoms was 2.3% and in those without reflux symptoms was 1.2% (P = .18). In those with esophagitis, the prevalence was 2.6%; in those without, the prevalence was 1.4% (P = .32). Alcohol (P = .04) and smoking (P = .047) were independent risk factors for BE. CONCLUSIONS: BE was found in 1.6% of the general Swedish population. Alcohol and smoking were significant risk factors. PMID- 16344052 TI - Computed tomographic virtual colonoscopy computer-aided polyp detection in a screening population. AB - BACKGROUND & AIMS: The sensitivity of computed tomographic (CT) virtual colonoscopy (CT colonography) for detecting polyps varies widely in recently reported large clinical trials. Our objective was to determine whether a computer program is as sensitive as optical colonoscopy for the detection of adenomatous colonic polyps on CT virtual colonoscopy. METHODS: The data set was a cohort of 1186 screening patients at 3 medical centers. All patients underwent same-day virtual and optical colonoscopy. Our enhanced gold standard combined segmental unblinded optical colonoscopy and retrospective identification of precise polyp locations. The data were randomized into separate training (n = 394) and test (n = 792) sets for analysis by a computer-aided polyp detection (CAD) program. RESULTS: For the test set, per-polyp and per-patient sensitivities for CAD were both 89.3% (25/28; 95% confidence interval, 71.8%-97.7%) for detecting retrospectively identifiable adenomatous polyps at least 1 cm in size. The false positive rate was 2.1 (95% confidence interval, 2.0-2.2) false polyps per patient. Both carcinomas were detected by CAD at a false-positive rate of 0.7 per patient; only 1 of 2 was detected by optical colonoscopy before segmental unblinding. At both 8-mm and 10-mm adenoma size thresholds, the per-patient sensitivities of CAD were not significantly different from those of optical colonoscopy before segmental unblinding. CONCLUSIONS: The per-patient sensitivity of CT virtual colonoscopy CAD in an asymptomatic screening population is comparable to that of optical colonoscopy for adenomas > or = 8 mm and is generalizable to new CT virtual colonoscopy data. PMID- 16344053 TI - Association of organic cation transporter risk haplotype with perianal penetrating Crohn's disease but not with susceptibility to IBD. AB - BACKGROUND & AIMS: Three years after the identification of NOD2/CARD15, 2 more genes for inflammatory bowel diseases (IBDs) were reported. The carnitine/organic cation transporter (OCTN) on 5q31 (IBD5) is associated with Crohn's disease (CD) and DLG5 (10q23), a member of membrane-associated guanylate kinase (MAGUK) family, with IBD. We studied mutation prevalence, assessed phenotypic expression, and performed conditional analysis to examine evidence for gene-gene interactions. METHODS: A cohort of 2032 individuals was genotyped for disease associated OCTN and DLG5 variants, including 981 patients with IBD (CD, n = 769; ulcerative colitis, n = 186; indeterminate colitis, n = 26) followed up at a tertiary IBD center. For 373 patients, DNA from both parents was available (cohort 1) for transmission disequilibrium testing analysis; case-control analysis was performed in 608 patients and 305 controls (cohort 2). RESULTS: There was no distortion of transmission toward affected offspring for any of the variant alleles. Case-control analysis also failed to shown an association. A higher frequency of DLG5 113A was observed in CARD15-positive patients (12.2%) compared with CARD15-negative patients (8.7%; P = .033). The OCTN-TC risk haplotype was associated with penetrating disease (odds ratio, 1.474; 95% confidence interval, 1.028-2.114; P = .035). For DLG5, there were no associations with a particular phenotype. CONCLUSIONS: DLG5 and OCTN do not play a role in the susceptibility to IBD, CD, or ulcerative colitis in the Flemish population but play a role in the phenotypic expression of the disease. OCTN variants were associated with perianal and penetrating CD. More studies in independent populations are urgently needed to assess the validity of DLG5 and OCTN in the pathogenesis of IBD. PMID- 16344054 TI - The contribution of OCTN1/2 variants within the IBD5 locus to disease susceptibility and severity in Crohn's disease. AB - BACKGROUND & AIMS: Recent data suggest that polymorphisms in the organic cation transporter (OCTN) genes OCTN1 (SLC22A4) and OCTN2 (SLC22A5) represent disease causing mutations within the IBD5 locus (chromosome 5q31). We investigated associations with disease susceptibility, phenotype, and evidence for epistasis with CARD15 in 679 patients with Crohn's disease (CD) or ulcerative colitis (UC). METHODS: A total of 374 patients with CD, 305 patients with UC, and 294 healthy controls (HCs) were studied. Genotyping for single nucleotide polymorphisms IGR2096, IGR2198, and IGR2230, OCTN1 variant (SLC22A4 1672C-->T), and OCTN2 variant (SLC22A5 -207G-->C) was performed using the TaqMan system. RESULTS: The IBD5 OCTN1 and OCTN2 polymorphisms were in strong linkage disequilibrium (D', >0.959). IGR2198 variant allele frequency (49.1% vs 40.8%; P = .0046) and homozygosity (21% vs 14.8%; P = .044) were associated with CD versus HCs. Variant allelic frequency of OCTN1 (53.6% vs 43%; P = .0008) and OCTN2 (56.1% vs 48.4%; P = .0092) polymorphisms and homozygosity for the OCTN1/2-TC haplotype (28.4% vs 16%; P = .0042) were associated with CD versus HCs. IGR2198 homozygosity and TC homozygosity were associated with stricturing/penetrating disease at follow-up (P = .011 and P = .011, respectively) and disease progression (P = .038 and P = .049, respectively) on univariate analysis and with need for surgery on multivariate analysis (P = .016 and P = .004, respectively). In the absence of the IBD5 risk haplotype, no association of OCTN1/2 variants with CD was detected. No associations were seen with UC. CONCLUSIONS: The IBD5 locus influences susceptibility, progression, and need for surgery in CD. However, the contribution of OCTN1/2 variants is not independent of the IBD5 haplotype; a causative role for these genes remains plausible but is not yet proven. Further genetic, functional, and expression data are now required. PMID- 16344055 TI - Risk of serious upper gastrointestinal toxicity with over-the-counter nonaspirin nonsteroidal anti-inflammatory drugs. AB - BACKGROUND & AIMS: Use of prescription nonaspirin nonsteroidal anti-inflammatory drugs (NANSAIDs) increases the risk of serious upper gastrointestinal toxicity. Less is known about over-the-counter (OTC) NANSAIDs, which are typically used at lower doses and for shorter durations. This study assessed the risk of toxicity with OTC NANSAIDs. METHODS: A total of 359 case subjects hospitalized for upper gastrointestinal bleeding, perforation, or benign gastric outlet obstruction were recruited from 28 hospitals. A total of 1889 control subjects were recruited by random digit dialing from the same region. Data on medication use were collected via structured telephone interview. RESULTS: Use of OTC NANSAIDs on > or = 4 days during the most recent week had an adjusted odds ratio (OR) of 1.83 (95% confidence interval [CI], 1.14-2.95). Use of high-dose OTC NANSAIDs during the index week had an adjusted OR of 5.21 (95% CI, 2.32-11.69). In contrast, use of OTC NANSAIDs <4 times during the index week (adjusted OR, 0.67; 95% CI, 0.43 1.06) and use of very low doses of prescription or OTC NANSAIDs during the index week (adjusted OR, 0.74; 95% CI, 0.49-1.12) were not significantly associated with an increased risk of serious gastrointestinal toxicity. We did not observe a significant difference between the risk of toxicity with OTC naproxen versus OTC ibuprofen (adjusted OR, 0.84; 95% CI, 0.26-2.70). CONCLUSIONS: Use of OTC NANSAIDs at recommended doses has a relatively good safety profile compared with prescription NANSAIDs. However, use of high-dose OTC NANSAIDs (comparable to a prescription dose) is associated with serious gastrointestinal toxicity. PMID- 16344056 TI - Clostridium difficile toxin A-induced colonocyte apoptosis involves p53-dependent p21(WAF1/CIP1) induction via p38 mitogen-activated protein kinase. AB - BACKGROUND & AIMS: Clostridium difficile toxin A causes marked apoptosis of colonocytes in vivo and in vitro, which contributes to the formation of ulcers and pseudomembranes. We investigated the role of p53-dependent pathways and p38 mitogen-activated protein kinase (p38) in toxin A-induced colonocyte apoptosis. METHODS: The effects of the activation of p53 and p53-dependent pathways including p21(WAF1/CIP1) were assessed in nontransformed human colonic NCM460 epithelial cells exposed to toxin A. Phosphorylation of p53 protein by p38 was measured by in vitro kinase assay, whereas p21 induction by activated p53 was determined by gel shift assays and RNA silencing (small interfering RNA). The relationship between colonocyte apoptosis and p38/p53-dependent pathways was studied in intact mice. RESULTS: Toxin A stimulated p38 and p53 activation and induced cell cycle arrest (G(2)-M) with persistent expression of p21(WAF1/CIP1). Blockage of p38 by SB203580 inhibited p53 phosphorylation and induction of p21(WAF1/CIP1). In intact mice, p38 blockade suppressed toxin A-mediated destruction of intestinal villi, p21(WAF1/CIP1) expression, and enterocyte apoptosis. In addition, toxin A-mediated p21(WAF1/CIP1) and Bak induction, cytochrome c release, and caspase-3 activation were markedly attenuated in p53 silenced colonocytes, despite active p38. Overexpression of p21(WAF1/CIP1) triggered apoptosis and increased toxin A-associated colonocyte apoptosis. CONCLUSIONS: The signaling pathway for colonocyte apoptosis following toxin A exposure involves p38-dependent activation of p53 and subsequent induction of p21(WAF1/CIP1), resulting in cytochrome c release and caspase-3 activation through Bak induction. PMID- 16344057 TI - Cancer characteristics in Swedish families fulfilling criteria for hereditary nonpolyposis colorectal cancer. AB - BACKGROUND & AIMS: The present study quantified the prevalence of families that fulfill the Amsterdam or Bethesda criteria for hereditary nonpolyposis colorectal cancer (HNPCC) in the whole Swedish population and investigated the extent to which tumors in the classified families are HNPCC-related. METHODS: The families of the Swedish Family-Cancer Database with at least 4 generations (N = 566,877) were classified according to the Amsterdam and the Bethesda criteria. Survival methods were used to assess the risk of cancer in the classified families, the prognosis of cancer patients, and the risk of subsequent malignancies after colorectal adenomas and after colorectal/endometrial adenocarcinomas. RESULTS: The Bethesda criteria identified 0.9% of all Swedish families and 11.2% of patients with colorectal cancer. Families that fulfilled the Bethesda criteria showed increased risks of cancer in the colorectum, endometrium, small bowel, ovary, stomach, bile ducts, renal pelvis, and ureter; members of Bethesda criteria families were at decreased risks of lung and cervical cancers. The prognosis of cancer in the ureter, renal pelvis, stomach, ovary, and colorectum, but not in the endometrium, was better in Bethesda criteria than in nonclassified families. CONCLUSIONS: Most malignancies in the classified families reflect typical features of HNPCC (association with subsequent malignancies, accelerated adenoma-carcinoma sequence, and better survival). The data presented in this study should help to define surveillance strategies for members of families that fulfill the criteria for HNPCC testing. PMID- 16344058 TI - The role of transsphincteric pressure and proximal gastric volume in acid reflux before and after fundoplication. AB - BACKGROUND & AIMS: This study was conducted to explore the role of proximal gastric volume and transsphincteric pressure on acid reflux during transient lower esophageal sphincter relaxation (TLESR) in patients with gastroesophageal reflux disease (GERD) both before and after laparoscopic fundoplication. METHODS: Twenty GERD patients were studied before and after fundoplication along with 20 healthy controls. High-resolution manometry and pH recording was performed for 1 hour before and 2 hours following meal ingestion (500 mL/300 kcal). Three dimensional ultrasonographic images of the stomach were acquired before and every 15 minutes after meal ingestion. RESULTS: Postprandial proximal to total gastric volume distribution ratios were significantly larger in GERD patients before fundoplication (0.57 +/- 0.01; P < .05) and smaller following fundoplication (0.37 +/- 0.01; P < .001) compared with controls (0.46 +/- 0.01). The percentage of TLESRs associated with acid reflux did not relate to proximal gastric volume in any subject group. The transsphincteric pressure profile was different for TLESRs with and without reflux in GERD patients as well as in healthy controls. The pressure gradient across the esophagogastric junction (EGJ) (DeltaEGJp) was greater for TLESRs with acid reflux compared with TLESRs without acid reflux in both GERD patients (11.4 +/- 0.8 vs 8.0 +/- 0.5 mm Hg, respectively; P < .01) and controls (10.6 +/- 0.7 vs 7.1 +/- 0.8 mm Hg, respectively; P < .05). After fundoplication, DeltaEGJp during TLESRs was 7.9 +/- 0.9 mm Hg, and the transsphincteric pressure profile markedly changed. CONCLUSIONS: Although proximal gastric volume plays a key role in eliciting TLESRs, it is not related to the incidence of acid reflux during TLESRs. The transsphincteric pressure gradient is greater in TLESRs associated with acid gastroesophageal reflux. PMID- 16344059 TI - A community screening program for Helicobacter pylori saves money: 10-year follow up of a randomized controlled trial. AB - BACKGROUND & AIMS: Population screening and treatment of Helicobacter pylori has been advocated as a means of reducing mortality from gastric cancer, as well as dyspepsia and dyspepsia-related resource use. Previous programs have failed to demonstrate a significant effect on mortality or resource use, but follow-up was only for 1 or 2 years. We aimed to determine the effect of screening for H pylori on dyspepsia and dyspepsia-related resource use over 10 years. METHODS: H pylori positive individuals, aged 40-49 years, enrolled in a community screening program, randomized to eradication therapy or placebo in 1994, were sent a validated dyspepsia questionnaire by mail 10 years later, and primary care records were reexamined. Consultation, referral, prescribing, and investigation data related to dyspepsia were extracted. United Kingdom costs were applied to derive total cost per person (1 pound = 1.8 dollars). RESULTS: Of 2324 original participants, 1864 (80%) were traced and contacted. Of these, 1086 (47%) responded, and 919 (40%) agreed to a review of their primary care records. There was a 10-year mean saving in total dyspepsia-related costs of 117 dollars per person (95% confidence interval [CI] = 11 dollars-220 dollars, P = .03) with eradication therapy. Those symptomatic at baseline showed a nonsignificant trend toward resolution of symptoms at 10 years with eradication therapy (relative risk of remaining symptomatic, 0.89; 95% CI: 0.77-1.03). CONCLUSIONS: There were significant reductions in total dyspepsia-related health care costs. The savings made were greater than the initial cost of H pylori screening and treatment. PMID- 16344060 TI - A new method for isolating colonocytes from naturally evacuated feces and its clinical application to colorectal cancer diagnosis. AB - BACKGROUND & AIMS: The early detection of colorectal cancer is desired because this cancer can be cured surgically if diagnosed early. The purpose of the present study was to determine the feasibility of a new methodology for isolating colonocytes from naturally evacuated feces, followed by cytology or molecular biology of the colonocytes to detect colorectal cancer originating from any part of the colorectum. METHODS: Several simulation studies were conducted to establish the optimal methods for retrieving colonocytes from any portion of feces. Colonocytes exfoliated into feces, which had been retrieved from 116 patients with colorectal cancer and 83 healthy volunteers, were analyzed. Part of the exfoliated colonocytes was examined cytologically, whereas the remainder was subjected to DNA analysis. The extracted DNA was examined for mutations of the APC, K-ras, and p53 genes using direct sequence analysis and was also subjected to microsatellite instability (MSI) analysis. RESULTS: In the DNA analysis, the overall sensitivity and specificity were 71% (82 of 116) of patients with colorectal cancer and 88% (73 of 83) of healthy volunteers. The sensitivity for Dukes A and B was 72% (44 of 61). Furthermore, the sensitivity for cancers on the right side of the colon was 57% (20 of 35). The detection rate for genetic alterations using our methodology was 86% (80 of 93) when the analysis was limited to cases in which genetic alterations were present in the cancer tissue. CONCLUSIONS: We have developed a new methodology for isolating colonocytes from feces. The present study describes a promising procedure for future clinical evaluations and the early detection of colorectal cancers, including right-side colon cancer. PMID- 16344061 TI - Coffee and tea consumption are associated with a lower incidence of chronic liver disease in the United States. AB - BACKGROUND & AIMS: Coffee drinking has been suggested to protect against liver injury, but it is uncertain whether this is of clinical significance. We examined the relationship of coffee and tea consumption with the incidence of hospitalization or death from chronic liver disease (CLD). METHODS: Participants in the population-based, first National Health and Nutrition Examination Survey, 1971-1975, were asked about coffee and tea consumption, which was categorized as <1 cup (mean, 0.2 cups), 1 to 2 cups, and >2 cups per day (mean, 4.0 cups). A second analysis included persons who, in 1982-1984, were asked more detailed questions on coffee and tea drinking. Participants were followed through 1992 1993 for a hospital or death certificate diagnosis of CLD or cirrhosis (ICD-9-CM 571). Hazard rate ratios for CLD according to coffee and tea intake were calculated using Cox proportional hazards analysis. RESULTS: Among 9849 persons followed for a median of 19.0 years (range, 0.02-22.1), the cumulative incidence of CLD was 1.4%. In multivariate analysis, participants who drank >2 cups per day had less than half the rate of CLD as those who drank <1 cup per day (hazard ratio, 0.43, 95% confidence interval: 0.24-0.78). Protection by coffee and tea was limited to persons at higher risk for liver diseases from heavier alcohol intake, overweight, diabetes, or high iron saturation. Among 9650 participants who provided detailed drink information in 1982-1984, intake of regular ground coffee and of caffeine was associated with lower incidence of CLD. CONCLUSIONS: Coffee and tea drinking decreases the risk of clinically significant CLD. PMID- 16344062 TI - Steatosis is a cofactor in liver injury in hemochromatosis. AB - BACKGROUND & AIMS: Obesity-related steatosis is an increasingly common histologic finding and often coexists with other chronic liver diseases. Although obesity and steatosis are recognized risk factors for more advanced fibrosis in chronic hepatitis C and alcoholic liver disease, it has not been determined whether these factors influence the progression of other diseases in which steatosis is not a feature of the primary liver insult. METHODS: We studied 214 patients with hemochromatosis who were homozygous for the C282Y substitution in HFE and had undergone liver biopsy prior to phlebotomy. RESULTS: Steatosis was present in 41.1% of these patients, and 14.5% had moderate or severe steatosis. Median serum alanine aminotransferase (ALT) and ferritin levels were higher (P < .001), and median transferrin saturation (P = .01) and hepatic iron concentration (HIC) were lower (P = .003) in subjects with steatosis compared with subjects without steatosis. Bivariate analysis revealed a significant association between steatosis and fibrosis (P = .001). Following multiple logistic regression, steatosis was independently associated with fibrosis (odds ratio [OR] 4.3, 95% confidence interval [CI]: 2.1-8.8; P < .001) along with male sex (OR, 5.1; 95% CI: 2.0-12.5; P < .001), excess alcohol consumption (males > or = 50 g/day, females > or = 40 g/day) (OR, 3.9; 95% CI: 1.8-8.5; P = .001), and hepatic iron content (OR, 1.4; 95% CI: 1.2-1.6; P < .001). Both higher BMI (OR, 3.3; 95% CI: 1.8-6.3; P < .001) and alcohol consumption (males > or = 30 g/day, females > or = 10 g/day) (OR, 3.4; 95% CI: 1.2-10.0; P = .023) were independently associated with the presence of steatosis. CONCLUSIONS: These findings indicate that obesity related steatosis may have a role as a cofactor in liver injury in hemochromatosis. This has important clinical implications and suggests that obesity should be actively addressed in the management of patients with hemochromatosis, as well as other liver diseases. PMID- 16344063 TI - Renal failure in patients with cirrhosis and sepsis unrelated to spontaneous bacterial peritonitis: value of MELD score. AB - BACKGROUND & AIMS: Although renal failure is a common complication of sepsis and patients with cirrhosis frequently develop sepsis, there have been no studies specifically assessing renal function in patients with cirrhosis and sepsis unrelated to spontaneous bacterial peritonitis. The aim of this study was to investigate prospectively the frequency, characteristics, and outcome of renal failure in patients with cirrhosis and sepsis unrelated to spontaneous bacterial peritonitis. METHODS: One hundred six consecutive patients with cirrhosis and sepsis were studied prospectively. Patients with spontaneous bacterial peritonitis were excluded. RESULTS: Twenty-nine out of 106 patients (27%) with cirrhosis and sepsis developed acute renal failure as compared with only 8 of 100 patients (8%) from a control group of cirrhotic patients without infection (P < .0001). Renal failure in the sepsis group was reversible in 22 (76%; 21% of all patients) patients and nonreversible in 7 (24%; 6% of all patients) patients. Renal failure was associated with impairment of effective arterial blood volume, without evidence of tubular damage. The occurrence and type of renal failure correlated strongly with mortality (mortality at 3 months: nonreversible renal failure, 100%; reversible renal failure, 55%; no renal failure, 13%). Among variables obtained at diagnosis of sepsis, the Model for End-Stage Liver Disease (MELD) score was the only independent predictive factor of mortality. CONCLUSIONS: Renal failure is common in patients with cirrhosis and sepsis unrelated to spontaneous bacterial peritonitis and is associated with arterial underfilling and renal vasoconstriction. Outcome is poor, even in the setting of reversible renal failure. The MELD score is the best prognostic marker of patients with cirrhosis and sepsis. PMID- 16344064 TI - Nitric oxide not carbon monoxide mediates nonadrenergic noncholinergic relaxation in the murine internal anal sphincter. AB - BACKGROUND & AIMS: Inhibitory reflexes in the internal anal sphincter (IAS) are controlled by inhibitory nonadrenergic, noncholinergic innervation (i-NANC). We investigated the roles of 3 different neurohumoral agonists as possible i-NANC neurotransmitters: carbon monoxide (CO), nitric oxide (NO), and vasoactive intestinal peptide (VIP). METHODS: IAS smooth muscle strips were isolated from wild-type (WT), heme oxygenase (HO)-2 knockout (HO-2-/-) and neuronal NO synthase (nNOS) knockout (nNOS-/-) mice. Relaxation of IAS was induced by CO, NO, VIP, and electrical field stimulation (EFS) in the presence and absence of neurohumoral inhibitors (tin protoporphyrin IX [SnPP IX] for CO synthesis, N(omega)-nitro-L arginine [L-NNA] for NO synthesis, and VIP(10-28) for VIP receptor). Western blot and immunohistochemistry were used to test the presence and localization of HO (for CO synthesis) types 1 (HO-1) and 2 (HO-2), neuronal NO synthase (nNOS, for NO synthesis), and VIP. RESULTS: All 3 neurohumoral agonists produced relaxation (with no difference between WT and HO-2-/- IAS), but CO was over 100 times less potent than NO and VIP. EFS produced relaxation in WT and HO-2-/- IAS with the same intensity. L-NNA and nNOS deletion (approximately 80%) and VIP(10-28) (approximately 15%) significantly inhibited the relaxations, whereas SnPP IX had no effect. Positive immunoreactivities for HO-2, nNOS, and VIP were found in the myenteric plexus of WT IAS. HO-2-/- IAS did not express immunoreactivity for HO 2. CONCLUSIONS: i-NANC relaxations of mouse IAS are primarily mediated via NO (by nNOS activity) and partly via VIP. CO directly relaxes the mouse IAS but does not play any significant role in the i-NANC relaxation. PMID- 16344065 TI - Cross-organ sensitization of lumbosacral spinal neurons receiving urinary bladder input in rats with inflamed colon. AB - BACKGROUND & AIMS: Clinical studies show that patients with irritable bowel syndrome and colonic diseases frequently experience sensory and motor dysfunctions of the urinary bladder. The aim of this study was to investigate the spinal neuronal mechanisms responsible for potential cross talk between these visceral organs. METHODS: Colonic inflammation was induced by dextran sulfate sodium (5%) in drinking water for 7-12 days (n = 12); another group of rats without dextran sulfate sodium (n = 12) was used as control. Extracellular potentials of single L6 to S2 spinal neurons were recorded in pentobarbital anesthetized and paralyzed rats with dextran sulfate sodium-induced colitis or normal colon. Urinary bladder distention (0.5-2.0 mL; 20 seconds) was produced with saline inflation, and colorectal distention was induced by inflation of an air balloon (80 mm Hg; 20 seconds). RESULTS: A total of 58 of 153 (38%) and 55 of 152 (36%) spinal neurons responded to urinary bladder distention in dextran sulfate sodium-treated and control animals, respectively. The mean background activity of neurons excited by urinary bladder distention in rats with dextran sulfate sodium-induced colitis was significantly higher than in the control group. The threshold volume for excitatory responses to urinary bladder distention in rats with inflamed colon (0.024 +/- 0.09 mL; n = 30) was significantly lower than for control rats (0.062 +/- 0.016 mL; n = 31; P < .05). The stimulus-response curves of excitatory responses to graded urinary bladder distention were significantly increased for both viscerovisceral (urinary bladder distention and colorectal distention) convergent neurons and urinary bladder distention-receptive neurons in rats with colitis compared with control animals. CONCLUSIONS: Acute colitis sensitized lumbosacral spinal neurons receiving input from the urinary bladder. Thus, spinal neuronal hyperexcitability may be involved in central cross-organ sensitization of visceral nociception between the colon and urinary bladder. PMID- 16344066 TI - Base treatment corrects defects due to misfolding of mutant cystic fibrosis transmembrane conductance regulator. AB - BACKGROUND & AIMS: The most common form of the disease-causing cystic fibrosis transmembrane conductance regulator mutation, DeltaF508, leads to a misfolded protein that undergoes endoplasmic reticulum-associated degradation. Retrieval of misfolded protein from the cis-Golgi or pre-Golgi intermediate compartment is a critical factor in endoplasmic reticulum retention and degradation of DeltaF508 protein. Therefore, the inhibition of retrograde Golgi-to-endoplasmic reticulum traffic by the alkalinization of Golgi lumen may permit functional DeltaF508 protein to reach the cell surface. METHODS: Functional and biochemical effects of alkaline treatment on misfolded cystic fibrosis transmembrane conductance regulator-induced defects were measured in CFPAC-1 cells, which endogenously express DeltaF508 cystic fibrosis transmembrane conductance regulator, and in CHO cells, which heterologously express DeltaF508 cystic fibrosis transmembrane conductance regulator. The animal survival rate and the functional expression of cystic fibrosis transmembrane conductance regulator proteins were analyzed in homozygous DeltaF508 mice after chronic treatment with weak base NaHCO3. RESULTS: In CFPAC-1 and CHO cells, intracellular alkalization by reducing carbon dioxide concentrations in a carbon dioxide incubation chamber or intra-Golgi alkalization by bafilomycin A1 treatment increased the membrane expression of DeltaF508 protein and cystic fibrosis transmembrane conductance regulator-dependent anion transport. Notably, chronic administration of NaHCO3 increased the long-term survival of homozygous DeltaF508 mice and induced the functional expression of cystic fibrosis transmembrane conductance regulator in the luminal membrane of intestinal epithelium. CONCLUSIONS: We found that base treatments correct misfolded cystic fibrosis transmembrane conductance regulator-induced defects in vitro and in vivo. These results imply that the alkalization of intracellular compartments, in particular, Golgi or pre-Golgi intermediate compartments, can be a potential therapeutic target for the loss-of-function type of conformational diseases. PMID- 16344067 TI - Targeted deletion of metalloproteinase 9 attenuates experimental colitis in mice: central role of epithelial-derived MMP. AB - BACKGROUND & AIMS: There is mounting evidence that matrix metalloproteinases are the predominant proteinases expressed in the gut mucosa during active inflammatory bowel disease. We investigated the role of metalloproteinase 9 (MMP 9), a secreted gelatinase that is consistently up-regulated in both animal models and human inflammatory bowel disease and is associated with disease severity, in the pathogenesis of colitis by using mice containing a targeted deletion of the MMP-9 gene. METHODS: Dextran sodium sulfate-induced colitis and Salmonella typhimurium-induced enterocolitis were used as animal models to study colitis. RESULTS: MMP-9 activity and protein expression were absent from normal colonic mucosa but were up-regulated during experimental colitis. MMP-9-/- mice exposed to dextran sodium sulfate or salmonella had a significantly reduced extent and severity of colitis. Immunohistochemical studies showed that MMP-9 was localized to epithelial cells and granulocytes during active colitis. The immune response to systemic administration of Salmonella typhimurium was not affected in MMP-9-/- mice. Neutrophil transmigration studies and bone marrow chimeras showed that neutrophil MMP-9 is neither required for its migration nor sufficient to induce tissue damage during colitis and that epithelial MMP-9 is important for tissue damage. MMP-9 inhibited cell attachment and wound healing in the model intestinal epithelial cell line, Caco2-BBE. CONCLUSIONS: Taken together, our data suggest that MMP-9 expressed by epithelial cells may play an important role in the development of colitis by modulating cell-matrix interaction and wound healing. Thus, strategies to inhibit MMP-9 may be of potential therapeutic benefit. PMID- 16344068 TI - Bile salt-induced apoptosis involves NADPH oxidase isoform activation. AB - BACKGROUND & AIMS: Hydrophobic bile salts trigger a rapid oxidative stress response as an upstream event of CD95 activation and hepatocyte apoptosis. METHODS: The underlying mechanisms were studied by Western blot, immunocytochemistry, protein knockdown, and fluorescence resonance energy transfer microscopy in rat hepatocytes and human hepatoma cell line 7 (Huh7). RESULTS: The rapid oxidative stress formation in response to taurolithocholate-3 sulfate (TLCS) was inhibited by diphenyleneiodonium, apocynin, and neopterin, suggestive for the involvement of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases. TLCS induced a rapid serine phosphorylation of the regulatory subunit p47phox, which was sensitive to inhibition of sphingomyelinase and protein kinase Czeta (PKCzeta). Inhibitors of p47phox phosphorylation and p47phox protein knockdown abolished the TLCS-induced oxidative stress response and blunted subsequent CD95 activation. Consequences of TLCS-induced oxidative stress were c-Jun-N-terminal kinase activation and Yes-dependent activation of the epidermal growth factor receptor (EGFR), followed by EGFR-catalyzed CD95 tyrosine phosphorylation, formation of the death-inducing signaling complex, and execution of apoptosis. As shown by fluorescence resonance energy transfer experiments in Huh7 cells, TLCS induced a c-Jun-N-terminal kinase-dependent EGFR/CD95 association in the cytosol and trafficking of this protein complex to the plasma membrane. Inhibition of EGFR tyrosine kinase activity by AG1478 allowed for cytosolic EGFR/CD95 association, but prevented targeting of the EGFR/CD95 complex to the plasma membrane. Both processes, and TLCS-induced Yes and EGFR activation, were sensitive to inhibition of sphingomyelinase, PKCzeta, or NADPH oxidases. CONCLUSIONS: The data suggest that hydrophobic bile salts activate NADPH oxidase isoforms with the resulting oxidative stress response triggering activation of the CD95 system and apoptosis. PMID- 16344069 TI - Agonist-induced vesiculation of the Golgi apparatus in pancreatic acinar cells. AB - BACKGROUND & AIMS: The pancreatic acinar cell is known to regulate exocytosis, total protein synthesis, and secretory protein transport in response to a secretory stimulus. Whether secretory vesicle formation also is regulated is unclear. In this study, we determined whether agonist stimulation induces morphologic alterations in the acinar cell Golgi apparatus, and we evaluated the role of the vesicle severing protein dynamin. METHODS: Changes in Golgi structural integrity by examining the distribution of various Golgi and TGN lipid and protein markers in live and fixed cells on stimulation with cholecystokinin were noted in a primary pancreatic acinar cell model. Multiple dynamin reagents were used to examine the distribution and function of this molecular pinchase in resting and stimulated cells. RESULTS: Regulated secretion in acinar cells induced (1) marked fragmentation of the trans-Golgi network (TGN) that corresponded temporally with an increase in cytoplasmic calcium whereas pre-TGN compartments of the Golgi and regions of the TGN involved in the generation of constitutively trafficking vesicles were unaffected by agonist, and (2) significant recruitment of dynamin to the acinar cell Golgi apparatus that appeared to potentiate fragmentation of the TGN. CONCLUSIONS: These results suggest that the TGN is a dynamic organelle that fragments in response to cholecystokinin stimulation, a process that may contribute to zymogen granule formation. PMID- 16344070 TI - erbB-2/neu transformed rat cholangiocytes recapitulate key cellular and molecular features of human bile duct cancer. AB - BACKGROUND & AIMS: Cholangiocarcinomas appear to arise from the malignant transformation of cholangiocytes lining the biliary tract. Because the development of an in vitro model of malignant transformation can provide a powerful new tool for establishing critical events governing the molecular pathogenesis of cholangiocarcinoma, we investigated the potential of achieving malignant transformation of cultured rat cholangiocytes in relation to aberrant overexpression of mutationally activated erbB-2/neu. METHODS: Malignant neoplastic transformation was achieved after infection of the rat cholangiocyte cell line, designated BDE1, with the retrovirus Glu664-neu, containing the transforming rat erbB-2/neu oncogene. RESULTS: Compared with untransformed control cells, malignant transformants carrying the activating erbB-2/neu mutation prominently overexpressed p185neu receptor protein, which was phosphorylated strongly at its major autophosphorylation site at tyrosine 1248. Moreover, erbB-2/neu transformation of BDE1 cells resulted in increased telomerase activity, up-regulation of cyclooxygenase-2 with overproduction of prostaglandin E(2), enhanced phosphorylation of mitogen-activated protein kinase and of serine/threonine kinase Akt/PKB, overexpression of vascular endothelial growth factor, and increased mucin 1 messenger RNA expression. Only erbB-2/neu transformants were tumorigenic when transplanted into isogeneic rats, yielding a 100% incidence of tumors closely resembling human desmoplastic ductal cholangiocarcinomas in their morphology. Malignant cholangiocytes in the tumors were strongly immunoreactive for biliary cytokeratin 19, p185neu, and cyclooxygenase-2. CONCLUSIONS: This unique malignant transformation model recapitulates key molecular features of the human disease and appears to be well suited for testing novel molecular therapeutic strategies against cholangiocarcinoma. PMID- 16344071 TI - Epstein-barr virus-related gastric adenocarcinoma: an early secondary cancer post hemopoietic stem cell transplantation. AB - BACKGROUND & AIMS: Epstein-Barr virus (EBV) infection has been associated with some cases of gastric cancer. METHODS: We studied a case of early onset gastric adenocarcinoma after nonmyeloablative hematopoietic stem cell transplantation for myeloma in a 56-year-old man. RESULTS: The development of gastric adenocarcinoma was preceded by severe graft-versus-host disease (GVHD) necessitating strong immunosuppression, which resulted in an intense reactivation of EBV infection. Three sequential gastric biopsy examinations performed at 100, 130, and 150 days after hematopoietic stem cell transplantation showed gastritis, dysplasia, and adenocarcinoma, respectively. There was no evidence of Helicobacter pylori infection. Quantitative polymerase chain reaction for circulating EBV showed a surge of EBV DNA peaking at the time of gastritis, followed by a gradual decrease afterward with adequate control of GVHD and tailing of immunosuppression. In situ hybridization for EBV-encoded early small RNA showed absence of EBV in the gastritis specimen, but the presence of EBV in the dysplastic and carcinoma specimens. Aberrant promoter methylation of E-cadherin was observed only in the carcinoma specimens, showing that infection with EBV preceded E-cadherin methylation. CONCLUSIONS: Mucosal damage caused by GVHD, immunosuppression, and EBV reactivation combined to lead to EBV infection of the gastric cells and initiation of carcinogenesis, suggesting this case to be a genuine EBV-related opportunistic malignancy post-transplantation. An interesting proposition is that this case also might reflect a compacted timeline of events in EBV-related gastric cancers developing in immunocompetent patients. PMID- 16344072 TI - Liver gene expression signature of mild fibrosis in patients with chronic hepatitis C. AB - BACKGROUND & AIMS: The molecular mechanisms of hepatocellular carcinoma have been studied, but little is known of the changes in liver gene expression during the different stages of chronic hepatitis C virus (HCV) infection, in particular the transition from mild to moderate fibrosis. METHODS: We used real-time quantitative RT PCR to study the messenger RNA expression of 240 selected genes in 2 pools of liver specimens according to the stages of fibrosis (Metavir score; mild fibrosis = F1 and septal fibrosis = F2). Genes whose expression differed between pools (F2 vs F1) by at least 2-fold were selected. In addition, the expression level of these selected genes then was assessed in each of the 62 individual samples (F4, n = 6; F3, n = 17; F2, n = 21; vs F1, n = 18). RESULTS: The 22 genes that were up-regulated in the 21 F2 samples relative to the 18 F1 samples mainly encoded genes involved in cytoskeleton (KRT 19 and SCG 10), growth factors/cytokines (CXCL6, interleukin 8 [IL8], IL1A, IL2, and CXCL10), or growth factor receptors (CCR2, CXCR3, and CXCR4), or were involved in extracellular matrix production (COL1A1, CHI3L, and SPP1), in extracellular matrix remodeling (TIMP1, MMP7, and MMP9), and in cell junction (ITGA2 and CLDN 4). When hierarchically clustering the F2 and F1 samples according to the expression of the 11 most discriminatory genes (KRT 19, COL1A1, STMN2, CXCL6, CCR2, TIMP1, IL8, IL1A, ITGA2, CLDN 4, and IL2), the patient population was categorized into 2 subgroups: F1 and F2. Specifically, 15 of 18 F1 (83%) and 19 of 21 F2 (90%) were classified correctly (P < 10(-5)). We also studied the messenger RNA expression of these 240 selected genes in normal liver in comparison with F1. Genes dysregulated in the transition from normal liver to F1 mainly were interferon inducible genes, and therefore were very different from those dysregulated in the transition from F1 to F2. CONCLUSIONS: Genes involved in extracellular matrix turnover and immune response are implicated in the transition from mild to moderate fibrosis. Eleven of the genes could form the basis for the gene expression signature of mild versus moderate fibrosis in patients with chronic hepatitis C. PMID- 16344074 TI - Neural stem cell therapy and gastrointestinal biology. PMID- 16344073 TI - Endogenous stimulators and inhibitors of angiogenesis in gastrointestinal cancers: basic science to clinical application. AB - Progression of cancer is dependent on acquisition of vascular networks within the tumor. Tumor angiogenesis is dependent on up-regulation of angiogenesis stimulators to overcome the endogenous anti-angiogenic barrier. Such disruption of angiogenesis balance to favor neovascularization is a key step for progression of tumor growth and metastasis. In this regard, the vascular basement membrane and the extracellular matrix have been found to be rich sources of angiogenesis stimulators and inhibitors that become bioavailable on proteolysis of the matrix by tumor microenvironment-related enzymes. In this review the subgroup of endogenous angiogenesis stimulators and inhibitors is discussed, and their mechanism of action during tumor angiogenesis is evaluated. The role in regulating tumor growth and the possibility of using them as prognostic markers for human gastrointestinal cancers is discussed. Furthermore, we specifically address the role of vascular endothelial growth factor in human gastrointestinal cancers and discuss the development and use of bevacizumab (Avastin; anti vascular endothelial growth factor antibody [Genentech, CA]) in the treatment of colorectal and other gastrointestinal cancers. PMID- 16344075 TI - Presidential address during the AGA Plenary Session. PMID- 16344076 TI - A population prevalence of Barrett's esophagus--finally. PMID- 16344077 TI - Progress in refining virtual colonoscopy for colorectal cancer screening. PMID- 16344078 TI - Understanding association and causality in the genetic studies of inflammatory bowel disease. PMID- 16344079 TI - Another way for NADPH oxidase to send a signal for liver injury. PMID- 16344080 TI - Chemotherapy-related steatohepatitis: proceed with caution. PMID- 16344081 TI - Adding fuel to the fire: GM-CSF for active Crohn's disease. PMID- 16344082 TI - Stellate cell/HCV interactions in hepatic fibrosis. PMID- 16344083 TI - Can histopathology be the "Gold Standard" for diagnosing autoimmune pancreatitis? PMID- 16344085 TI - Chemoprevention of hepatocellular carcinoma in chronic hepatitis B with lamivudine? PMID- 16344087 TI - Is the incidence of esophageal adenocarcinomas in patients with Barrett's esophagus really overestimated? PMID- 16344090 TI - Systematic review of endoscopic therapy for ulcers with clots: Can a meta analysis be misleading? PMID- 16344091 TI - Stool DNA: a viable option for colorectal cancer screening. PMID- 16344092 TI - Natural variation in toxicity of wheat. PMID- 16344096 TI - Lenalidomide and thalidomide: an evolving paradigm for the management of multiple myeloma. PMID- 16344097 TI - Treatment paradigms for the newly diagnosed patient with multiple myeloma. AB - The management of newly diagnosed multiple myeloma is currently being revolutionized by the emergence of novel therapeutic agents. Thalidomide, the thalidomide analogue lenalidomide, and the proteasome inhibitor bortezomib each have potential roles to play in the initial therapy of myeloma that are supported by encouraging clinical trials results. While evaluation for stem cell transplantation (SCT) remains a part of the algorithm for management of the newly diagnosed patient, these drugs offer improved options for induction before, and possibly maintenance after, SCT. For the patient who is not a candidate for SCT the armamentarium for management has been significantly expanded. This review will summarize currently available data regarding these exciting agents and will describe therapeutic strategies currently on the horizon. PMID- 16344098 TI - Lenalidomide: patient management strategies. AB - Experience with thalidomide has underscored the need for diligent monitoring and management of side effects to achieve optimal therapeutic benefit. Initial phase II and III results with lenalidomide, a thalidomide analogue, have confirmed impressive response rates with a significantly more favorable toxicity profile. The Cleveland Clinic experience with lenalidomide alone or in combination with pegylated liposomal doxorubicin, vincristine, and reduced schedule dexamethasone will be described. Peripheral neuropathy and deep venous thrombosis (DVT). Low dose aspirin alleviated DVT when lenalidomide was used in combination with anthracycline-containing chemotherapy. Strategies for managing these toxicities and for recognition and management of lenalidomide-associated myelosuppression are reviewed. PMID- 16344099 TI - Lenalidomide and thalidomide: mechanisms of action--similarities and differences. AB - Multiple myeloma is a B-cell malignancy characterized by an excess of monotypic plasma cells in the bone marrow. The molecular mechanisms that are involved in disease progression depend on the interaction between the multiple myeloma cells and the bone microenvironment. Because these mechanisms have been well characterized, it is possible to develop regimens that are more specific to pathways involved in the pathogenesis of multiple myeloma than is typical for conventional chemotherapy in disease management. Thalidomide and immunomodulatory drugs (IMiDs) have now been shown to block several pathways important for disease progression in multiple myeloma. First established as agents with antiangiogenic properties, thalidomide and IMiDs inhibit the production of interleukin (IL)-6, which is a growth factor for the proliferation of myeloma cells. In addition, they activate apoptotic pathways through caspase 8-mediated cell death. At the mitochondrial level, they are responsible for c-jun terminal kinase (JNK) dependent release of cytochrome-c and Smac into the cytosol of cells, where they regulate the activity of molecules that affect apoptosis. By activating T cells to produce IL-2, thalidomide and IMiDs alter natural killer (NK) cell numbers and function, thus augmenting the activity of NK-dependent cytotoxicity. Data delineating these events have been derived from experiments done in resistant and sensitive multiple myeloma cell lines. Although thalidomide and IMiDs demonstrate similar biologic activities, IMiDs are more potent than thalidomide and achieve responses at lower doses. Lenalidomide, a thalidomide derivative, has also been shown to have a different toxicity profile. Our understanding of the mechanism of action of these agents has provided a platform for exciting clinical trials evaluating combinations of thalidomide and lenalidomide with both conventional chemotherapy and newer targeted agents. PMID- 16344100 TI - Management of the relapsed/refractory myeloma patient: strategies incorporating lenalidomide. AB - The immunomodulatory drug (IMiD) lenalidomide is a more potent immunomodulator than thalidomide with respect to its effects on cytokine modulation and increased T-cell proliferation. Of all the IMiDs, clinical trial data are most mature for lenalidomide. In phase I studies, dose-limiting toxicities of lenalidomide were limited to myelosuppression and a response rate of 72% was seen in relapsed/refractory patients. Three phase II studies subsequently evaluated the efficacy of single-agent lenalidomide or lenalidomide in combination with dexamethasone. As a single agent for post-transplant salvage therapy, lenalidomide 25 mg every 3 weeks has shown response rates as high as 44%. For patients with relapsed/refractory multiple myeloma, the MM-007 study has shown that lenalidomide alone or in combination with dexamethasone provides response rates between 37% and 41%. In MM-007, median progression-free survival was 5.5 months at early analysis and the median overall survival has yet to be reached. Preliminary data for the single-arm, multicenter, open-label MM-014 study showed that median time to progression was 5.6 months. Response rates indicate that 70% of patients had stable disease or better as the best response to treatment. Two randomized, phase III trials (MM-009 and MM-010) evaluated lenalidomide with high dose dexamethasone versus high-dose dexamethasone alone for the treatment of relapsed/refractory multiple myeloma. Both MM-009 and MM-010 provided remarkably similar response rates for patients receiving lenalidomide and dexamethasone. In both trials response rates with the combination were greater than twice the response rates seen with high-dose dexamethasone alone. Indeed, an independent Data Monitoring Committee determined that both trials exceeded the prespecified efficacy value of P<.0015, recommending that the trials be discontinued and that lenalidomide be offered to patients on the dexamethasone arm of the trial if clinically indicated. Toxicities observed in studies of lenalidomide alone were low; the incidence of peripheral neuropathy was significantly lower than those noted in trials using thalidomide. Thrombocytopenia was a significant grade 3 or 4 toxicity observed; however, it was manageable with dose reduction. In contrast with high-dose dexamethasone, deep vein thrombosis has emerged as an important toxicity. Lenalidomide is currently being tested in combination with both standard and novel agents, including bortezomib, for patients with relapsed/refractory multiple myeloma. PMID- 16344101 TI - The choroid plexus-cerebrospinal fluid system: from development to aging. AB - The function of the cerebrospinal fluid (CSF) and the tissue that secretes it, the choroid plexus (CP), has traditionally been thought of as both providing physical protection to the brain through buoyancy and facilitating the removal of brain metabolites through the bulk drainage of CSF. More recent studies suggest, however, that the CP-CSF system plays a much more active role in the development, homeostasis, and repair of the central nervous system (CNS). The highly specialized choroidal tissue synthesizes trophic and angiogenic factors, chemorepellents, and carrier proteins, and is strategically positioned within the ventricular cavities to supply the CNS with these biologically active substances. Through polarized transport systems and receptor-mediated transcytosis across the choroidal epithelium, the CP, a part of the blood-CSF barrier (BCSFB), controls the entry of nutrients, such as amino acids and nucleosides, and peptide hormones, such as leptin and prolactin, from the periphery into the brain. The CP also plays an important role in the clearance of toxins and drugs. During CNS development, CP-derived growth factors, such as members of the transforming growth factor-beta superfamily and retinoic acid, play an important role in controlling the patterning of neuronal differentiation in various brain regions. In the adult CNS, the CP appears to be critically involved in neuronal repair processes and the restoration of the brain microenvironment after traumatic and ischemic brain injury. Furthermore, recent studies suggest that the CP acts as a nursery for neuronal and astrocytic progenitor cells. The advancement of our knowledge of the neuroprotective capabilities of the CP may therefore facilitate the development of novel therapies for ischemic stroke and traumatic brain injury. In the later stages of life, the CP-CSF axis shows a decline in all aspects of its function, including CSF secretion and protein synthesis, which may in themselves increase the risk for development of late-life diseases, such as normal pressure hydrocephalus and Alzheimer's disease. The understanding of the mechanisms that underlie the dysfunction of the CP-CSF system in the elderly may help discover the treatments needed to reverse the negative effects of aging that lead to global CNS failure. PMID- 16344102 TI - Zebrafish genetics and formation of embryonic vasculature. AB - The embryonic vasculature develops in a conserved manner in all vertebrates. Endothelial progenitor cells differentiate from mesodermal cells, then migrate and assemble into the dorsal aorta and the cardinal vein. This primitive circulatory loop undergoes sprouting and branching via a two-step navigation mechanism to form the trunk vascular network. Various studies using several model systems have uncovered a number of signaling mechanisms that regulate these complex processes. A genetic approach in zebrafish has led to identification of mutations and molecules that are responsible for specification of endothelial progenitor cells, differentiation of arterial and venous cells, and patterning of the dorsal aorta and intersegmental vessels. These studies highlight the unique utilities and benefits of the zebrafish system for studying development of embryonic blood vessels. PMID- 16344103 TI - Leaf senescence: signals, execution, and regulation. AB - Leaf senescence is a type of postmitotic senescence. The onset and progression of leaf senescence are controlled by an array of external and internal factors including age, levels of plant hormones/growth regulators, and reproductive growth. Many environmental stresses and biological insults such as extreme temperature, drought, nutrient deficiency, insufficient light/shadow/darkness, and pathogen infection can induce senescence. Perception of signals often leads to changes in gene expression, and the upregulation of thousands of senescence associated genes (SAGs) causes the senescence syndrome: decline in photosynthesis, degradation of macromolecules, mobilization of nutrients, and ultimate cell death. Identification and analysis of SAGs, especially genome-scale investigations on gene expression during leaf senescence, make it possible to decipher the molecular mechanisms of signal perception, execution, and regulation of the leaf senescence process. Biochemical and metabolic changes during senescence have been elucidated, and potential components in signal transduction such as receptor-like kinases and MAP kinase cascade have been identified. Studies on some master regulators such as WRKY transcription factors and the senescence-responsive cis element of the senescence-specific SAG12 have shed some light on transcriptional regulation of leaf senescence. PMID- 16344104 TI - Muscle stem cells and regenerative myogenesis. AB - A population of myogenic progenitors termed satellite cells undertakes postnatal development and repair of skeletal muscle. Studies have indicated that atypical myogenic precursors can also participate in muscle regeneration. The source of this regenerative capacity has been attributed to "adult stem cells" that represent poorly understood multipotent cell lineages, believed to reside in all adult tissue populations. Here we review the origin and location of muscle satellite cells and stem cells, as well as the mechanisms by which they may be specified. We discuss how the experimental models utilized raise important questions regarding the validity of extrapolating these findings. PMID- 16344105 TI - Gene regulation in spermatogenesis. AB - Mammalian spermatogenesis is a complex hormone-dependent developmental program in which a myriad of events must take place to ensure that germ cells reach their proper stage of development at the proper time. Many of these events are controlled by cell type- and stage-specific transcription factors. The regulatory mechanisms involved provide an intriguing paradigm for the field of developmental biology and may lead to the development of new contraceptives an and innovative routs to treat male infertility. In this review, we address three aspects of the genetic regulatory mechanism that drive spermatogenesis. First, we detail what is known about how steroid hormones (both androgens and estrogens) and their cognate receptors initiate and maintain mammalian spermatogenesis. Steroids act through three mechanistic routes: (i) direct activation of genes through hormone dependent promoter elements, (ii) secondary transcriptional responses through activation of hormone-dependent transcription factors, and (iii) rapid, transcription-independent (nonclassical) events induced by steroid hormones. Second, we provide a survey of transcription factors that function in mammalian spermatogenesis, including homeobox, zinc-finger, heat-shock, and cAMP-response family members. Our survey is not intended to cover all examples but to give a flavor for the gamut of biological roles conferred by transcription factors in the testis, particularly those defined in knockout mice. Third, we address how testis-specific transcription is achieved. In particular, we cover the evidence for and against the idea that some testis-specific genes are transcriptionally silent in somatic tissues as a result of DNA methylation. PMID- 16344106 TI - Modeling age-related diseases in Drosophila: can this fly? AB - Human neurodegenerative diseases are characterized by progressive neuronal cell loss often resulting in memory and cognitive decline, motor dysfunction, and ultimately premature death. Despite the prevalence of these diseases, there are no effective cures. Insight into many of these syndromes has come from the identification of single gene mutations that are associated with inherited forms of the disease. This has led to the development of animal models in which the pathogenesis caused by these genes can be rigorously examined. Due to their short life span and powerful genetic potential, several attempts have been made to model neurodegenerative diseases in the fruit fly Drosophila melanogaster. This review will describe how these models were generated and how faithfully they recapitulate human disease. In addition, how fly models can be used to identify genetic modifiers of known disease genes and what these have revealed about the biochemical pathways underlying disease pathogenesis is discussed. Finally, the review will describe how fly models can be used to identify new therapeutic targets and test the effectiveness of new drugs. PMID- 16344107 TI - Cell death and organ development in plants. AB - Programmed cell death (PCD) is an important feature of plant development; however, the mechanisms responsible for its regulation in plants are far less well understood than those operating in animals. In this review data from a wide variety of plant PCD systems is analyzed to compare what is known about the underlying mechanisms. Although senescence is clearly an important part of plant development, only what is known about PCD during senescence is dealt with here. In each PCD system the extracellular and intracellular signals triggering PCD are considered and both cytological and molecular data are discussed to determine whether a unique model for plant PCD can be derived. In the majority of cases reviewed, PCD is accompanied by the formation of a large vacuole, which ruptures to release hydrolytic enzymes that degrade the cell contents, although this model is clearly not universal. DNA degradation and the activation of proteases is also common to most plant PCD systems, where they have been studied; however, breakdown of DNA into nucleosomal units (DNA laddering) is not observed in all systems. Caspase-like activity has also been reported in several systems, but the extent to which it is a necessary feature of all plant PCD has not yet been established. The trigger for tonoplast rupture is not fully understood, although active oxygen species (AOS) have been implicated in several systems. In two systems, self incompatibility and tapetal breakdown as a result of cytoplasmic male sterility, there is convincing evidence for the involvement of mitochondria including release of cytochrome c. However, in other systems, the role of the mitochondrion is not clear-cut. How cells surrounding the cell undergoing PCD protect themselves against death is also discussed as well as whether there is a link between the eventual fate of the cell corpse and the mechanism of its death. PMID- 16344108 TI - The blood-testis barrier: its biology, regulation, and physiological role in spermatogenesis. AB - The blood-testis barrier (BTB) in mammals, such as rats, is composed of the tight junction (TJ), the basal ectoplasmic specialization (basal ES), the basal tubulobulbar complex (basal TBC) (both are testis-specific actin-based adherens junction [AJ] types), and the desmosome-like junction that are present side-by side in the seminiferous epithelium. The BTB physically divides the seminiferous epithelium into basal and apical (or adluminal) compartments, and is pivotal to spermatogenesis. Besides its function as an immunological barrier to segregate the postmeiotic germ-cell antigens from the systemic circulation, it creates a unique microenvironment for germ-cell development and confers cell polarity. During spermatogenesis, the BTB in rodents must physically disassemble to permit the passage of preleptotene and leptotene spermatocytes. This occurs at late stage VII through early stage VIII of the epithelial cycle. Studies have shown that this dynamic BTB restructuring to facilitate germ-cell migration is regulated by two cytokines, namely transforming growth factor-beta3 (TGF-beta3) and tumor necrosis factor-alpha (TNFalpha), via downstream mitogen-activated protein kinases. These cytokines determine the homeostasis of TJ- and basal ES structural proteins, proteases, protease inhibitors, and other extracellular matrix (ECM) proteins (e.g., collagen) in the seminiferous epithelium. Some of these molecules are known regulators of focal contacts between the ECM and other actively migrating cells, such as macrophages, fibroblasts, or malignant cells. These findings also illustrate that cell-cell junction restructuring at the BTB is regulated by mechanisms involved in the junction turnover at the cell-matrix interface. This review critically discusses these latest findings in the field in light of their significance in the biology and regulation of the BTB pertinent to spermatogenesis. PMID- 16344109 TI - Angiogenic factors in the pathogenesis of preeclampsia. AB - Preeclampsia affects 5-10% of pregnancies and is responsible for substantial maternal and neonatal morbidity and mortality. It is believed to be a two-stage disease with an initial placental trigger with no maternal symptoms followed by a maternal syndrome characterized by hypertension, proteinuria, and endothelial dysfunction. The first stage is thought to be due to shallow cytotrophoblast invasion of maternal spiral arterioles leading to placental insufficiency. The diseased placenta in turn releases soluble angiogenic factors that induce systemic endothelial dysfunction and clinical preeclampsia during the second stage. This review will discuss the role of circulating angiogenic factors of placental origin as potential mediators of the systemic endothelial dysfunction and the clinical syndrome of preeclampsia and provide an evolutionary explanation for this phenomenon. PMID- 16344110 TI - Cellulose metabolism in plants. AB - Many bacterial genomes contain a cellulose synthase operon together with a cellulase gene, indicating that cellulase is required for cellulose biosynthesis. In higher plants, there is evidence that cell growth is enhanced by the overexpression of cellulase and prevented by its suppression. Cellulase overexpression could modify cell walls not only by trimming off the paracrystalline sites of cellulose microfibrils, but also by releasing xyloglucan tethers between the microfibrils. Mutants for membrane-anchored cellulase (Korrigan) also show a typical phenotype of prevention of cellulose biosynthesis in tissues. All plant cellulases belong to family 9, which endohydrolyzes cellulose, but are not strong enough to cause the bulk degradation of cellulose microfibrils in a plant body. It is hypothesized that cellulase participates primarily in repairing or arranging cellulose microfibrils during cellulose biosynthesis in plants. A scheme for the roles of plant cellulose and cellulases is proposed. PMID- 16344111 TI - Survivin: a protein with dual roles in mitosis and apoptosis. AB - Survivin is a fascinating little protein that acts as a component of the chromosomal passenger complex, which is essential for cell division, and as an inhibitor of apoptosis. With dual roles in promoting cell proliferation and preventing apoptosis, it is considered a protein that interfaces life and death. Interest in survivin has been fueled by its abundance in human cancers, where it has potential as a prognostic marker for cancer, and as a target for chemotherapy. Accordingly, since its discovery in 1997, publications on survivin have risen exponentially in basic and clinical fields alike. This review highlights the key advances in our understanding of the cellular function of this protein. PMID- 16344112 TI - Hypothalamic tanycytes: a key component of brain-endocrine interaction. AB - Tanycytes are bipolar cells bridging the cerebrospinal fluid (CSF) to the portal capillaries and may link the CSF to neuroendocrine events. During the perinatal period a subpopulation of radial glial cells differentiates into tanycytes, a cell lineage sharing some properties with astrocytes and the radial glia, but displaying unique and distinct morphological, molecular, and functional characteristics. Four populations of tanycytes, alpha(1,2) and beta(1,2), can be distinguished. These subtypes express differentially important functional molecules, such as glucose and glutamate transporters; a series of receptors for neuropeptide and peripheral hormones; secretory molecules such as transforming growth factors, prostaglandin E(2), and the specific protein P85; and proteins of the endocytic pathways. This results in functional differences between the four subtypes of tanycytes. Thus, alpha(1,2) tanycytes do not have barrier properties, whereas beta(1,2) tanycytes do. Different types of tanycytes use different mechanisms to internalize and transport cargo molecules; compounds internalized via a clathrin-dependent endocytosis would only enter tanycytes from the CSF. There are also differences in the neuron-tanycyte relationships; beta(1,2) tanycytes are innervated by peptidergic and aminergic neurons, but alpha(1,2) tanycytes are not. Important aspects of the neuron-beta(1) tanycyte relationships have been elucidated. Tanycytes can participate in the release of gonadotropin releasing hormone (GnRH) to the portal blood by expressing estrogen receptors, absorbing molecules from the CSF, and providing signal(s) to the GnRH neurons. Removal of tanycytes prevents the pulse of GnRH release into the portal blood, the peak of luteinizing hormone, and ovulation. The discovery in tanycytes of new functional molecules is opening a new field of research. Thus, thyroxine deiodinase type II, an enzyme generating triiodothyronine (T(3)) from thyroxine, appears to be exclusively expressed by tanycytes, suggesting that these cells are the main source of brain T(3). Glucose transporter-2 (GLUT-2), a low-affinity transporter of glucose and fructose, and ATP-sensitive K(+) channels are expressed by tanycytes, suggesting that they may sense CSF glucose concentrations. PMID- 16344113 TI - Short retroposons in eukaryotic genomes. AB - Short retroposons (SINEs) are repetitive elements amplified in the genome via an RNA intermediate, using the enzymatic machinery of autonomous retroposons (LINEs). SINEs are widely distributed in eukaryotes; for instance, all tested mammalian genomes contain 10(4)-10(6) SINE copies. Although several SINE families such as primate Alu or rodent B1 have long been recognized, the more recent discovery of many SINEs in various eukaryotes, as well as progress in understanding the mechanisms of LINE replication and genome functioning as a whole, shed light on the biology and evolution of SINEs and their significance for the cell. PMID- 16344114 TI - Plant genome analysis: the state of the art. AB - Plants are the basis for the survival of all "higher" organisms on Earth. Development of molecular genetics tools has allowed analysis of the structure, evolution, and function of whole plant genomes, rather than individual genes. DNA based markers were instrumental in constructing detailed genetic maps of model plants and all major crop species. These molecular maps were the basis of physical maps and the first plant whole genome sequences. Comparative analysis based on genetic, cytogenetic, and physical maps and DNA sequence information provided new insights into the evolution of plant nuclear and organellar genomes. Mapping factors controlling Mendelian and quantitative traits made possible the cloning and functional characterization of novel genes, which function in plant development, adaptation to biotic and abiotic stress, or in the formation of other agronomic characters. The parallel analysis of all transcripts, proteins, and metabolites present in plant cells or tissues has generated information that may lead to a better integrated understanding of genome function. Postfunctional analysis of natural variation of gene function and its effects on phenotype is envisaged to provide new diagnostic and therapeutic molecular tools for applications in plant breeding, adaptation, and ecology. PMID- 16344115 TI - Role of Pex19p in the targeting of PMP70 to peroxisome. AB - Pex19p is a protein required for the peroxisomal membrane synthesis. The 70-kDa peroxisomal membrane protein (PMP70) is synthesized on free cytosolic ribosomes and then inserted posttranslationally into peroxisomal membranes. Pex19p has been shown to play an important role in this process. Using an in vitro translation system, we investigated the role of Pex19p as a chaperone and identified the regions of PMP70 required for the interaction with Pex19p. When PMP70 was translated in the presence of purified Pex19p, a large part of PMP70 existed as soluble form and was co-immunoprecipitated with Pex19p. However, in the absence of Pex19p, PMP70 formed aggregates during translation. To identify the regions that interact with Pex19p, various truncated PMP70 were translated in the presence of Pex19p and subjected to co-immunoprecipitation. The interaction was markedly reduced by the deletion of the NH(2)-terminal 61 amino acids or the region around TMD6. Further, we expressed these deletion constructs of PMP70 in fusion with the green fluorescent protein in CHO cells. Fusion proteins lacking these Pex19p binding sites did not display any peroxisomal localization. These results suggest that Pex19p binds to PMP70 co-translationally and keeps PMP70 as a proper conformation for the localization to peroxisome. PMID- 16344116 TI - HIV infection is associated with increased NTPDase activity that correlates with CD39-positive lymphocytes. AB - Infection with the human immunodeficiency virus (HIV) results in alterations in immune cells such as an increase or decrease of cytokine secretion and immunodeficiency. HIV causes a state of chronic cellular activation that can induce apoptosis in lymphocyte T-helpers, making the patient susceptive to opportunistic infections. The biochemical mechanisms involved in this immune response to HIV have been researched. Here, we have shown for the first time that ATP and ADP hydrolysis are essential for the immune response to HIV. Our results clearly indicate an increase of NTPDase-1 (EC 3.6.1.5) activity in lymphocytes of HIV-positive patients, confirmed by an enhanced CD39 expression on its surface. These results suggest that NTPDase-1 may be important to keep an adequate balance between the generation and consumption of ATP and to preserve cellular integrity and immune response to the HIV infection. PMID- 16344117 TI - Oxidative modification of IkappaB by monochloramine inhibits tumor necrosis factor alpha-induced NF-kappaB activation. AB - We have previously reported that monochloramine (NH(2)Cl), a neutrophil-derived oxidant, inhibited tumor necrosis factor alpha (TNFalpha)-induced expression of cell adhesion molecules and nuclear factor-kappaB (NF-kappaB) activation (Free Radical Research 36 (2002) 845-852). Here, we studied the mechanism how NH(2)Cl inhibited TNFalpha-induced NF-kappaB activation, and compared the effects with taurine chloramine (Tau-NHCl). Pretreatment of Jurkat cells with NH(2)Cl at 70 microM resulted in suppression of TNFalpha-induced IkappaB phosphorylation and degradation, and inhibited NF-kappaB activation. In addition, a slow-moving IkappaB band appeared on SDS-PAGE. By contrast, Tau-NHCl for up to 200 microM had no effects. Interestingly, NH(2)Cl did not inhibit IkappaB kinase activation by TNFalpha. Protein phosphatase activity did not show apparent change. When recombinant IkappaB was oxidized by NH(2)Cl in vitro and phosphorylated by TNFalpha-stimulated Jurkat cell lysate, its phosphorylation occurred less effectively than non-oxidized IkappaB. In addition, when NF-kappaB-IkappaB complex was immunoprecipitated from NH(2)Cl-treated cells and phosphorylated in vitro by recombinant active IkappaB kinase, native IkappaB but not oxidized IkappaB was phosphorylated. Amino acid analysis of the in vitro oxidized IkappaB showed methionine oxidation to methionine sulfoxide. Although Tau-NHCl alone had little effects on TNFalpha-induced NF-kappaB activation, simultaneous presence of Tau-NHCl and ammonium ion significantly inhibited the NF-kappaB activation, probably through the conversion of Tau-NHCl to NH(2)Cl. These results indicated that NH(2)Cl inhibited TNFalpha-induced NF-kappaB activation through the oxidation of IkappaB, and that NH(2)Cl is physiologically more relevant than Tau NHCl in modifying NF-kappaB-mediated cellular responses. PMID- 16344119 TI - Prediction of birth weight by ultrasound in Turkish population. Which formula should be used in Turkey to estimate fetal weight? AB - To determine optimal sonographic fetal weight estimation formula for male and female infants, a prospective study has been performed. Uncomplicated pregnancies and 465 newborns were evaluated. Measurements included birth weight, length and head circumference in addition to fetal head circumference, femur length, abdominal circumference and biparietal diameter. Actual weights were compared with estimated weights calculated by ten different formula. Estimated fetal weight obtained from all formula, except those of Merz, Warsof and Ferrero, tended to be lower than the measured birth weight. The smallest mean differences were obtained with Hadlock 1, Hadlock 2, Hadlock 4 and Shepard formula (19 g-85 g), whereas Merz and Woo produced largest mean differences (110 g-364 g). Intraclass correlation coefficients (ICCs) generated with Hadlock 1 and Hadlock 2 formula were identical (0.90). ICCs obtained with Hadlock 3 and Hadlock 4, Shepard, Merz, Warsof and Campbell formula varied between 0.84 and 0.88. Hadlock 1 and 2 formula gave the closest approximation of birth weight in Turkish population. PMID- 16344118 TI - The Cas family docking protein, HEF1, promotes the formation of neurite-like membrane extensions. AB - The Cas family proteins are a family of adhesion docking molecules that mediate protein-protein interactions and contribute to a number of signal transduction pathways. Recent studies of two family members, p130Cas and Sin, have suggested that they may play a role in neurite formation. The current study demonstrates that the third family member, HEF1, can also stimulate the formation of neurite like processes, in the presence of Rho kinase inhibitors. The HEF1-promoted processes actively extend from the cell body and resemble neurites both in the manner of process extension and in the distribution of adhesion-associated molecules. The HEF1-promoted processes are dependent on the presence of an intact microtubule system and can be inhibited by co-expression of either constitutively active Rac or Cdc42 GTPase. Together, our data support a role for the Cas proteins in regulating cellular morphologies that contribute to tissue specialization. PMID- 16344120 TI - Ultrasonographic characterization of the uterine artery in the nonestrus bitch. AB - The canine species is often used as an experimental animal model in medicine. Doppler ultrasound of uterine arteries has important clinical implications in obstetrics and gynecology in women and animal species. To our knowledge, the ultrasonographic features of uterine arteries in the bitch have not been reported. Thus, an ultrasonographic technique to locate the uterine arteries in the bitch is described in this study. A total of 11 nonspayed nonestrus bitches underwent abdominal ultrasound examination. Pulsed-wave, color and power Doppler settings were used to detect the location and characterize the flow of the uterine arteries. Mean values for systolic peak velocity, diastolic peak velocity, diastolic reversal flow and resistive index were calculated. Uterine arteries had a high pulsatility flow in all cases. Future work is needed to fully assess the clinical importance of the uterine artery flow in the bitch, as well as to study possible waveform changes at different estrus phases and throughout gestation. PMID- 16344122 TI - Optimization of factor analysis of the left ventricle in echocardiography for detecting wall motion abnormalities. AB - This study has tested solutions to optimize the ability of factor analysis of the left ventricle in echocardiography (FALVE) to detect segmental wall motion abnormalities automatically. On four- and two-chamber views of 38 patients, two factors (one flat curve and one curve describing the contraction-relaxation sequence) were extracted and associated factor images were combined to synthesize a parametric image (constant image in green, positive/negative values of the contraction-relaxation image in red/blue). The segments were graded on the visual and the parametric views. The impact of selecting a whole cardiac cycle, masking the left ventricle and realigning the image sequence on the results, was demonstrated. Systematic realignment had a positive impact, especially for patients with left bundle branch block or pacemaker. After alignment, for the entire population, the absolute concordance was 68.6% and the relative concordance (within one grade) was 99%. Thus, FALVE is promising for detecting segmental wall motion abnormalities. PMID- 16344121 TI - Diabetes and diastolic function: stiffness and relaxation from transmitral flow. AB - To characterize the mechanism by which diabetes affects the heart in diabetic (n = 15) and age-matched control subjects (n = 15), we quantified and compared diastolic function (DF) in terms of chamber stiffness and viscosity/relaxation by analyzing Doppler E- and E'-waves and simultaneous (high-fidelity) hemodynamic data. We compared tau, standard Doppler indexes and indexes of stiffness and viscosity/relaxation computed via the parameterized diastolic filling (PDF) formalism. Three PDF parameters uniquely characterize each E-wave in terms of load (x(o)), viscoelasticity or viscosity/relaxation (c) and stiffness (k). Significant differences for c (p = 0.00004), the peak atrioventricular pressure gradient (kx(o)) (p = 0.02) and the stored elastic energy available for early filling (1/2kx(o)2) (p = 0.04) were found. The only conventional index attaining significance was E-wave acceleration time (p = 0.007). Neither time constant of isovolumic relaxation (tau) nor E-wave deceleration time, E', k or x(o) differentiated between groups. We conclude that PDF based DF assessment differentiates between diabetic and nondiabetic controls better than conventional echo- or cath-based indexes. Our results in humans agree with published results from animal studies. We conclude that diabetes affects the heart via a quantifiable increase in chamber viscoelasticity (c) rather than an increase in chamber stiffness (k) and that phenotypic characterization of diabetic cardiomyopathy is facilitated by DF assessment via the PDF formalism. PMID- 16344123 TI - 3-D measurement of body tissues based on ultrasound images with 3-D spatial information. AB - In this study, we developed a new method to perform 3-D measurements between the recorded B-scans using the corresponding spatial location and orientation of each B-scan, without the need to create a 3-D volume. A portable ultrasound (US) scanner and an electromagnetic spatial locator attached to the US probe were used. During data collection, the US probe was moved over the region-of-interest. A small number of B-scans containing interesting anatomical information were captured from different body parts and displayed in a 3-D space with their corresponding locations recorded by the spatial locator. In the B-scan planes, the distance between any two points, as well as the angle between any two lines, could be calculated. In validation experiments, three distances and three angles of a custom-designed phantom were measured using this method. In comparison with the results measured by a micrometer, the mean error of distance measurement was 0.8 +/- 1.7 mm (-2.3 +/- 3.6%) and that of angle measurement was -0.3 +/- 2.9 degrees (-0.1 +/- 4.1%). The lengths of the first metatarsals and the angles between the first metatarsals and the middle part of the tibias of three subjects were measured in vivo using magnetic resonance imaging (MRI) and the US method by two operators before and after MRI scanning. The overall percentage differences of the length and angle measurements were 0.8 +/- 2.2% and 2.5 +/- 3.6%, respectively. The results showed that this US method had good repeatability and reproducibility (interclass correlation coefficient values > 0.75). We expect that this new method could potentially provide a quick and effective approach for the 3-D measurement of soft tissues and bones in the musculoskeletal system. PMID- 16344124 TI - Comparison of phalangeal ultrasound and dual energy X-ray absorptiometry in healthy male and female adolescents. AB - The aims of this study were to determine if there is a correlation between dual energy X-ray absorptiometry (DXA) and phalangeal quantitative ultrasound (QUS) in identifying children and adolescents with low bone density, and to assess if body size influences the results of the two techniques to the same degree. Measurements were performed in 67 girls and 83 boys aged 14 to 19 y using DBM Sonic 1200 (IGEA, Carpi, Italy) and the DXA equipment (LUNAR Radiation Corp., Madison, WI, USA). Twelve adolescents (eight males and four females) reported a past history of nonosteoporotic fractures. Lumbar spine bone mineral density (LS BMD), total body bone mineral density (TB BMD) and total body bone mineral content (TB BMC) correlated positively with age, height, BMI and weight, in both genders. Amplitude-dependent speed of sound (Ad-SOS) was positively correlated with age, height and Tanner stages in both genders and negatively correlated with BMI in females. TB BMD, TB BMC and LS BMD positively correlated with Ad-SOS only in males. In females, there were no significant correlations between Ad-SOS, TB BMD, TB BMC and LS BMD measurements. Twelve teenagers with previous fractures (high impact fractures) were found to have lower DXA and QUS values than age matched teenagers without fractures but the statistical significance was found only in relation to TB BMD values (p = 0.02). In conclusion, we obtained results similar to those that have been reported by other authors using different QUS techniques. Furthermore, the Ad-SOS measurements taken at the distal metaphysis of the proximal phalanges correlate poorly with LS BMD and TB BMD measured by DXA in growing subjects. PMID- 16344125 TI - Anisotropy of apparent backscatter in the short-axis view of mouse hearts. AB - The goals of this investigation were to measure the anisotropy of backscattered ultrasound observed in the short-axis view of mouse hearts in systole and diastole and to compare these measurements with predictions from a computer simulation. Measurements of midmyocardial apparent backscatter were obtained from analyses of the hearts of seven wild-type mice using a clinical imaging system utilizing a linear array with a nominal center frequency of 13 MHz. A computer model simulating the short-axis view was implemented based on previous measurements of the angle of insonification dependence of myocardial backscatter and attenuation. Results demonstrate that the measured backscatter was largest for those myocardial regions corresponding to approximately perpendicular insonification relative to the myofibers and the smallest for regions of approximately parallel insonification, with the minimum to maximum values of apparent backscatter differing by approximately 10 dB. The measured anisotropy of backscatter was similar for end-systole and end-diastole and was in good agreement with the predicted anisotropy obtained from the computer simulations. PMID- 16344126 TI - Robustness of reconstructing the Young's modulus distribution of vulnerable atherosclerotic plaques using a parametric plaque model. AB - Assessment of atherosclerotic plaque composition is crucial for quantitative monitoring of atherosclerosis and for quantifying the effect of pharmaceutical plaque-stabilizing treatments during clinical trials. We assessed this composition by applying a geometrically constrained, iterative inverse solution method to reconstruct a modulus elastogram (i.e., Young's modulus image) from a plaque strain elastogram (i.e., radial strain image) that is measured using intravascular ultrasound strain elastography. This reconstruction method is especially suited for thin-cap fibroatheromas (TCFAs) (i.e., plaques with a thin fibrous cap overlaying a lipid pool). Because a strain elastogram of a plaque depends upon the plaque material composition, catheter position within the vessel and measurement noise, this paper investigates how robust the reconstruction is when these parameters are varied. To this end, a standard plaque was defined as the modulus elastogram that was reconstructed from an in vivo measured strain elastogram of a human coronary plaque. This standard plaque was used to computer simulate different strain elastograms, by varying the 1. geometry and material properties of its plaque components, 2. catheter position and 3. level of added strain noise. Robustness was evaluated by quantifying the correctly reconstructed size, shape and Young's modulus of each plaque component region and minimal cap thickness. The simulations showed that TCFAs can be adequately reconstructed; the thinner and stiffer the cap or the softer and larger the lipid pool, the better is the reconstruction of these components and minimal cap thickness. Furthermore, reconstructions were 1. independent of catheter position and 2. independent of strain noise. As such, this method has potential to monitor robustly and quantitatively atherosclerosis in vivo. PMID- 16344127 TI - Cell-competition algorithm: a new segmentation algorithm for multiple objects with irregular boundaries in ultrasound images. AB - Segmentation of multiple objects with irregular contours and surrounding sporadic spots is a common practice in ultrasound image analysis. A new region-based approach, called cell-competition algorithm, is proposed for simultaneous segmentation of multiple objects in a sonogram. The algorithm is composed of two essential ideas. One is simultaneous cell-based deformation of regions and the other is cell competition. The cells are generated by two-pass watershed transformations. The cell-competition algorithm has been validated with 13 synthetic images of different contrast-to-noise ratios and 71 breast sonograms. Three assessments have been carried out and the results show that the boundaries derived by the cell-competition algorithm are reasonably comparable to those delineated manually. Moreover, the cell-competition algorithm is robust to the variation of regions-of-interest and a range of thresholds required for the second-pass watershed transformation. The proposed algorithm is also shown to be superior to the region-competition algorithm for both types of images. PMID- 16344128 TI - The effects of residual temperature rise on ultrasound heating. AB - In recent theoretical studies, the temperature rise produced by diagnostic ultrasound was estimated by solving the Bioheat Transfer Equation (BHTE) but ignoring the initial temperature rise. The temperature rise was determined in our study by the BHTE including an initial temperature rise. We discuss how the initial temperature rise occurs during an ultrasound examination, and how the initial temperature rise affects subsequent ultrasound heating. We theoretically show that the temperature rise produced by the ultrasound examination (exposure time of 500 s) in a tissue sample having an initial temperature rise was higher than that in a tissue sample with no initial temperature rise that was exposed to ultrasound (exposure time of 1200 s). The theoretical results for these two cases were 5.64 degrees C and 3.58 degrees C, respectively. In our experimental study, the highest temperature rise was measured in the presence of an initial temperature rise as in the theoretical study under the same exposure conditions. Mean temperature rises for tissue without an initial temperature rise and for tissue with an initial temperature rise were 2.42 +/- 0.13 degrees C and 3.62 +/- 0.17 degrees C, respectively. Both theoretical and experimental studies show that unless the initial temperature rise produced by the first ultrasound examination decreases to 0 degrees C, the next ultrasound examination on the same tissue sample may cause the temperature rise to be higher than expected. PMID- 16344129 TI - Investigation of intensity thresholds for ultrasound tissue erosion. AB - Our previous studies have shown that short intense pulses delivered at certain pulse repetition frequencies (PRF) can achieve localized, clean erosion in soft tissue. In this paper, the intensity thresholds for ultrasound induced erosion and the effects of pulse intensity on erosion characterized by axial erosion rate, perforation area and volume erosion rate were investigated on in vitro porcine atrial wall tissue. Ultrasound pulses with a 3-cycle pulse duration and a 20-kHz PRF were delivered by a 788-kHz single element focused transducer. I(SPPA) values of 1000 to 9000 W/cm2 were tested. Results show the following: (1) the estimated intensity threshold for generating erosion was at I(SPPA) of 3220 W/cm2; (2) the axial erosion rate increased with higher intensity at I(SPPA) < or = 5000 W/cm2, while decreased with higher intensity at I(SPPA) > or = 5000 W/cm2; and (3) the perforation area and the volume erosion rate increased with higher intensity. PMID- 16344130 TI - Delivery of oligodeoxynucleotides into human saphenous veins and the adjunct effect of ultrasound and microbubbles. AB - Therapy with naked oligodeoxynucleotides (ODNs, molecular weight: 3000 to 7500) provides an elegant means of modulating gene expression without the problems associated with conventional gene therapy, but the relatively low transfer efficiency on intravascular administration is a limitation to clinical application. Ultrasound, which can be potentiated by microbubbles, shows promise as a method of delivering macromolecules such as plasmid DNA and other transgenes into cells. Since uptake of molecules into cells depends on their molecular weight, it might be expected that the delivery of ODNs, which are relatively small, will be facilitated by ultrasound and microbubbles. In the present study, we delivered ODNs into veins using ultrasound and microbubbles. First, we quantified the uptake of fluorescent-labeled ODNs into intact ex vivo human saphenous veins and isolated smooth muscle cells from the veins, evaluating the effect of ultrasound and microbubbles on uptake. Ultrasound potentiated the delivery of ODN in cells, except at high concentrations. When intact veins were studied, we achieved nuclear localization of fluorescent-labeled ODNs in cells. This increased with increasing concentration and incubation time and was not potentiated by ultrasound, even when microbubbles were used. We then applied a therapeutic ODN (antisense to intercellular adhesion molecule 1, ICAM-1) to vein samples and documented a functional inhibition of gene expression in a sequence specific manner at the protein level with immunohistochemistry and western blot analysis. Again, no significant difference was seen with adjunct ultrasound. These observations suggest high diffusion of ODNs into human saphenous veins in this ex vivo model, indicating potential applications to inhibition of vascular bypass graft occlusion and other vasculopathies. Although microbubble-ultrasound was of value with cells in culture, it was not beneficial with intact veins. PMID- 16344131 TI - Sonic activation of molecularly-targeted nanoparticles accelerates transmembrane lipid delivery to cancer cells through contact-mediated mechanisms: implications for enhanced local drug delivery. AB - Liquid perfluorocarbon nanoparticles serve as sensitive and specific targeted contrast and drug delivery vehicles by binding to specific cell surface markers. We hypothesized that application of acoustic energy at diagnostic power levels could promote nanoparticle-associated drug delivery by stimulating increased interaction between the nanoparticle's lipid layer and the targeted cell's plasma membrane. Ultrasound (mechanical index = 1.9) applied with a conventional ultrasound imaging system to nanoparticles targeted to alpha(v)beta3-integrins on C32 melanoma cancer cells in vitro produced no untoward effects. Within 5 min, lipid delivery from nanoparticles into cell cytoplasm was dramatically augmented. We also demonstrate the operation of a potential physical mechanism for this effect, the acoustic radiation force on the nanoparticles, which may contribute to the enhanced lipid delivery. Accordingly, we propose that local delivery of lipophilic substances (e.g., drugs) from targeted nanoparticles directly into cell cytoplasm can be augmented rapidly and safely with conventional ultrasound imaging devices through nondestructive mechanisms. PMID- 16344132 TI - The effect of low-intensity pulsed ultrasound on repair of epithelial cell monolayers in vitro. AB - Low intensity pulsed ultrasound (LIPUS) is widely used to accelerate tissue regeneration following injury, but the biological mechanisms of this effect are poorly understood. An in vitro model of epithelial wound healing was used to investigate the effect of LIPUS on the reepithelialization of scrape wounds in normal human urothelial (NHU) cell monolayers. The effects of clinical doses of ultrasound treatment on NHU cell growth and migration were investigated in cells grown under optimal conditions, without growth supplements and in media containing low vs. physiological calcium concentrations. No differences in cell growth or migration were observed. We conclude that there is no direct effect upon uro-epithelial regeneration by therapeutic ultrasound in vitro and suggest that any stimulation of epithelial wound repair in vivo may occur indirectly, for example by modulating the extracellular matrix composition and/or production of paracrine factors by the stroma. PMID- 16344133 TI - Synergistic effect of ultrasound and phthalocyanines on nucleated erythrocytes in vitro. AB - The synergistic effect of ultrasound (US) and chemicals on cells is known as sonodynamic therapy. In this work, two phthalocyanines (zinc and chloroaluminum) have been tested as potential sonosensitizers for sonodynamic therapy. We studied the effect of US and phthalocyanines on carp erythrocytes, as a nucleated cell model. The level of hemolysis, osmotic fragility, lipid peroxidation and oxidation of hemoglobin were the markers of these reactions. Red blood cells were preincubated with phthalocyanines and exposed to 1 MHz continuous wave at the intensity of 2.44 W/cm2 for 5 min. It was noticed that US and phthalocyanines exposure led to an increase in the level of hemolysis, lipid peroxidation product and osmotic fragility in comparison to US alone and phthalocyanines alone. However, these factors did not cause changes in the degree of hemoglobin oxidation. The results lead to the conclusion that phthalocyanines caused synergistic effect with US, and it can be used as a sonosensitizer for sonodynamic therapy, but the mechanism of this action is still unclear. PMID- 16344134 TI - Transforming growth factor-beta1 mediates the effects of low-intensity pulsed ultrasound in chondrocytes. AB - Low-intensity pulsed ultrasound (LIPUS) has been shown to accelerate fracture healing, but the precise mechanism is still unknown. We used aggregate chondrocyte culture system to analyze LIPUS-induced effects on chondrocytes. First, Northern analyses revealed that LIPUS maintained higher expression levels of type II collagen and aggrecan mRNA and delayed the appearance of type X collagen mRNA expression. We also showed that DNA content was increased and that alkaline phosphatase activity was maintained low by daily treatment. Moreover, LIPUS significantly promoted transforming growth factor (TGF)-beta1 mRNA expression and the protein production at 2 h and 12 h after the treatment, respectively. Furthermore, recombinant TGF-beta1 protein mimicked the LIPUS effect and anti-TGF-beta1 neutralizing antibody reversed all these changes induced by the LIPUS treatment. These results indicate that LIPUS promotes the proliferation and retains the differentiation state of chondrocytes in the aggregate culture and that TGF-beta1 plays an important role in mediating the LIPUS effects in chondrocytes. PMID- 16344135 TI - Broadband attenuation and nonlinear propagation in biological fluids: an experimental facility and measurements. AB - The design and construction of a versatile experimental facility for making measurements of the frequency-dependence of attenuation coefficient (over the range 1 MHz to 25 MHz) and nonlinear propagation in samples of biological fluids is described. The main feature of the facility is the ability to perform all of the measurements on the same sample of fluid within a short period of time and under temperature control. In particular, the facility allows the axial development of nonlinear waveform distortion to be measured with a wideband bilaminar polyvinylidene difluoride membrane hydrophone to study nonlinear propagation in biological fluids. The system uses a variable length bellows to contain the fluid, with transparent Mylar end-windows to couple the acoustic field into the fluid. Example results for the frequency-dependence of attenuation of Dow Corning 200/350 silicone fluid, used as a standard fluid, are presented and shown to be in good agreement with alternative measurements. Measurements of finite amplitude propagation in amniotic fluid, urine and 4.5% human albumin solutions at physiological temperature (37 degrees C) are presented and compared with theoretical predictions using existing models. The measurements were made using a 2.25-MHz single-element transducer coupled to a polymethyl methacrylate lens with a focal amplitude gain of 12 in water. The transducer was driven with an eight-cycle tone burst at source pressures up to 0.137 MPa. In general, given an accurate knowledge of the medium parameters and source conditions, the agreement with theoretical prediction is good for the first five harmonics. PMID- 16344136 TI - Real-time software processing and audio reproduction of directional Doppler signals. AB - In ultrasound Doppler systems, directional signals are typically obtained by processing quadrature demodulated data with dedicated analog or digital circuits. In this paper, a software approach is proposed, that allows fully exploiting the reproduction and recording capabilities of low-cost personal computer sound cards and/or embedded chips. Forward/reverse signals are separated through a wideband Hilbert filter. No limitations are imposed on the input signal sample rate, which is matched to the standard output format of sound cards through a band-limited interpolation filter controlled in a feedback loop. The digital audio streaming is performed in real-time in a Windows-based application. The processed data are in a standard format compatible with real-time recording in waveform or compressed files, as requested in many research applications. Simulations and in vivo tests show a typical cross talk of -50 dB between forward and reverse components, with low latency time (39 ms) and central processing unit load compatible to currently available personal computers. PMID- 16344137 TI - Theodor W. Blackstad (1925-2003): a pioneer in quantitative neuroanatomy. PMID- 16344138 TI - Pioneering a golden age of cerebral microcircuits: the births of the combined Golgi-electron microscope methods. AB - Theodor W. Blackstad devised methods by which the synaptic connectivity of neuron somata and their dendritic and axonal processes in the CNS could be analyzed by the combined use of light and electron microscope techniques. His first publication on that subject dates from 1965 and was contemporary to the independent research by William K. Stell. The Golgi method was an obvious neuronal marker at those times, and Blackstad and Stell showed that the Golgi precipitate is electron-dense and intracellular and, therefore, it could help identify in the electron microscope, with great accuracy, profiles of neurons initially visualized in light microscopy. Besides this convergent research, Blackstad demonstrated for the first time that anterograde axonal degeneration could be combined with the Golgi-electron microscope method, allowing the identification of the neurons whose dendritic or somatic profiles were postsynaptic to the severed axonal afferent projections. Last, but not least, Blackstad pioneered de-impregnation techniques for electron microscopy of Golgi preparations. This had a great impact in the study of synaptic circuitry. The present account is a remembrance of the events that linked these early attempts with the development of a de-impregnation method based on gold toning by Alan Peters and the present author. PMID- 16344139 TI - Microglia response and P2 receptor participation in oxygen/glucose deprivation induced cortical damage. AB - In the present work, we used a unique cortical/striatal/subventricular zone organotypic model in order to analyze the role of resident microglia in oxygen/glucose deprivation and to check the presence and modulation of several P2 receptors in the cortex. Immunofluorescence with the microglial marker OX42 and pharmacological experiments with indomethacin indicate that activation and recruitment of microglia after the insult is linked to cellular loss, mainly in the cortex. The confocal analysis with OX42 shows that, among the P2 receptors tested, P2X4, and P2X7 are expressed on microglia, while P2X1 and P2Y(1-2-12), although present in the slices, did not co-localize, whereas P2X6 is not detected. The upregulation of P2X4 and P2X7 on microglia and the toxic effect that different P2 agonists exert on cortical slices during oxygen/glucose deprivation indicate that a purinergic mechanism is related to the microglia activity; the protective effect of the P2 antagonist TNP-ATP is also described. In order to better understand the relationship between P2 receptors and OGD activated microglia, we induced oxygen/glucose deprivation in co-cultures of organotypic slices and N9 microglia cell line. The presence of the N9 (which expresses P2X4 and P2X7 protein) in the cultures increases the damage in the cortex by 40% and the use of P2 antagonist PPADS reduced the cell damage due to the N9 activation. Our results show that microglia recruitment after a metabolic impairment is associated with cellular loss and that P2X4 and P2X7, are involved in microglia activity. The neuroprotective action exerted by TNP-ATP and PPADS and the possible use of purinergic antagonist in the pharmacological treatment of oxygen/glucose deprivation is also addressed. PMID- 16344140 TI - Anatomical and gene expression mapping of the ventral pallium in a three dimensional model of developing human brain. AB - Combining gene expression data with morphological information has revolutionized developmental neuroanatomy in the last decade. Visualization and interpretation of complex images have been crucial to these advances in our understanding of mechanisms underlying early brain development, as most developmental processes are spatially oriented, in topologically invariant patterns that become overtly distorted during brain morphogenesis. It has also become clear that more powerful methodologies are needed to accommodate the increasing volume of data available and the increasingly sophisticated analyses that are required, for example analyzing anatomy and multiple gene expression patterns at individual developmental stages, or identifying and analyzing homologous structures through time and/or between species. Three-dimensional models have long been recognized as a valuable way of providing a visual interpretation and overview of complex morphological data. We have used a recently developed method, optical projection tomography, to generate digital three-dimensional models of early human brain development. These models can be used both as frameworks, onto which normal or experimental gene expression data can be mapped, and as objects, within which topological morphological relationships can be investigated in silico. Gene expression patterns and selected morphological structures or boundaries can then be visualized individually or in different combinations in order to study their respective morphogenetic significance. Here, we review briefly the optical projection tomography method, placing it in the context of other methods used to generate developmental three dimensional models, and show the definition of some CNS anatomical domains within a Carnegie stage 19 human model. We also map the telencephalic EMX1 and PAX6 gene expression patterns to this model, corroborating for the first time the existence of a ventral pallium primordium in the telencephalon of human embryos, a distinct claustroamygdaloid histogenetic area comparable to the recently defined mouse primordium given that name [Puelles L, Kuwana E, Puelles E, Bulfone A, Shimamura K, Keleher J, Smiga S, Rubenstein JLR (2000) Pallial and subpallial derivatives in the embryonic chick and mouse telencephalon, traced by the expression of the genes Dlx-2, Emx-1, Nkx-2.1, Pax 6, and Tbr-1. J Comp Neurol 424:409-438; Puelles L, Martinez S, Martinez-de-la Torre M, Rubenstein JLR (2004) Gene maps and related histogenetic domains in the forebrain and midbrain. In: The rat nervous system, 3rd ed (Paxinos G, ed), pp 3 25. San Diego: Academic Press]. PMID- 16344141 TI - Moving up or moving down? Malpositioned cerebellar unipolar brush cells in reeler mouse. AB - Cerebellar morphogenesis occurs through a complex interplay of cell proliferation and migration that in mouse and rat begins about midgestation and ends in the third postnatal week. Cerebellar cells derive from germinative matrices in the ventricular zone and the external granular layer. Like granule cells, unipolar brush cells (UBCs) are excitatory interneurons situated in the granular layer of the cortex and innervated by mossy fibers. While granule cells are produced from the external granular layer, the generation of UBCs is still controversial. We utilized the reeler mutant mouse, which has widespread misplacement of neurons due to lack of Reelin protein, to ascertain the origin of UBCs. In the reeler cerebellum, which is small and lacks foliation, Purkinje cells are greatly reduced in number and in large part are located ectopically in deep cerebellar masses. Granule cells are also reduced in number and form an irregular granule cell layer. In this study we demonstrate that the reeler mutation influences the positioning of UBCs and also significantly reduces their number. Both subsets of UBCs identified in normal mouse, the calretinin-positive and the metabotropic glutamate receptor 1alpha-positive subsets, are affected in the reeler. About 40% of the calretinin-positive UBCs are ectopically situated in the deep cerebellar regions and the immediate vicinity of the ependyma of the fourth ventricle. Ectopic UBCs have discrete, although somewhat looser brushes than granular layer UBCs, but form synaptic junctions with complex axon terminals, possibly belonging to mossy fibers and UBC axons, like their normally situated counterpart. The observed displacement of UBCs in the reeler suggests that they originate from the ventricular zone. PMID- 16344142 TI - Specificity of antibodies: unexpected cross-reactivity of antibodies directed against the excitatory amino acid transporter 3 (EAAT3). AB - Specific antibodies are essential tools for identifying individual proteins in biological samples. While generation of antibodies is often straightforward, determination of the antibody specificity is not. Here we illustrate this by describing the production and characterization of antibodies to excitatory amino acid transporter 3 (EAAT3). We synthesized 13 peptides corresponding to parts of the EAAT3 sequence and immunized 6 sheep and 30 rabbits. All sera were affinity purified against the relevant immobilized peptide. Antibodies to the peptides were obtained in almost all cases. Immunoblotting with tissue extracts from wild type and EAAT3 knockout animals revealed that most of the antibodies did not recognize the native EAAT3 protein, and that some recognized other proteins. Several immunization protocols were tried, but strong reactions with EAAT3 were only seen with antibodies to the C-terminal peptides. In contrast, good antibodies were obtained to several parts of EAAT2. EAAT3 was only detected in neurons. However, rabbits immunized with an EAAT3-peptide corresponding to residues 479-498 produced antibodies that labeled axoplasm and microtubules therein particularly strongly. On blots, these antibodies recognized both EAAT3 and a slightly smaller, but far more abundant protein that turned out to be tubulin. The antibodies were fractionated on columns with immobilized tubulin. One fraction contained antibodies apparently specific for EAAT3 while another fraction contained antibodies recognizing both EAAT3 and tubulin despite the lack of primary sequence identity between the two proteins. Addition of free peptide to the incubation solution blocked immunostaining of both EAAT3 and tubulin. CONCLUSIONS: Not all antibodies to synthetic peptides recognize the native protein. The peptide sequence is more important than immunization protocol. The specificity of an antibody is hard to predict because cross-reactivity can be specific and to unrelated molecules. The antigen preabsorption test is of little value in testing the specificity of affinity purified antibodies. PMID- 16344143 TI - New techniques for imaging, digitization and analysis of three-dimensional neural morphology on multiple scales. AB - Cognitive impairment in normal aging and neurodegenerative diseases is accompanied by altered morphologies on multiple scales. Understanding of the role of these structural changes in producing functional deficits in brain aging and neuropsychiatric disorders requires accurate three-dimensional representations of neuronal morphology, and realistic biophysical modeling that can directly relate structural changes to altered neuronal firing patterns. To date however, tools capable of resolving, digitizing and analyzing neuronal morphology on both local and global scales, and with sufficient throughput and automation, have been lacking. The precision of existing image analysis-based morphometric tools is restricted at the finest scales, where resolution of fine dendritic features and spine geometry is limited by the skeletonization methods used, and by quantization errors arising from insufficient imaging resolution. We are developing techniques for imaging, reconstruction and analysis of neuronal morphology that capture both local and global structural variation. To minimize quantization error and evaluate more precisely the fine geometry of dendrites and spines, we introduce a new shape analysis technique, the Rayburst sampling algorithm that uses the original grayscale data rather than the segmented images for precise, continuous radius estimation, and multidirectional radius sampling to represent non-circular branch cross-sections and anisotropic structures such as dendritic spine heads, with greater accuracy. We apply the Rayburst technique to 3D neuronal shape analysis at different scales. We reconstruct and digitize entire neurons from stacks of laser-scanning microscopy images, as well as globally complex structures such as multineuron networks and microvascular networks. We also introduce imaging techniques necessary to recover detailed information on three-dimensional mass distribution and surface roughness of amyloid beta plaques from human Alzheimer's disease patients and from the Tg2576 mouse that expresses the "Swedish" mutation of the amyloid precursor protein. By providing true three-dimensional morphometry of complex histologic structures on multiple scales, the tools described in this report will enable multiscale biophysical modeling studies capable of testing potential mechanisms by which altered dendritic structure, spine geometry and network branching patterns that occur in normal aging and in many brain disorders, determine deficits of functions such as working memory and cognition. PMID- 16344144 TI - Database and tools for analysis of topographic organization and map transformations in major projection systems of the brain. AB - Integration of dispersed and complicated information collected from the brain is needed to build new knowledge. But integration may be hampered by rigid presentation formats, diversity of data formats among laboratories, and lack of access to lower level data. We have addressed some of the fundamental issues related to this challenge at the level of anatomical data, by producing a coordinate based digital atlas and database application for a major projection system in the rat brain: the cerebro-ponto-cerebellar system. This application, Functional Anatomy of the Cerebro-Cerebellar System in rat (FACCS), is available via the Rodent Brain WorkBench (http://www.rbwb.org). The data included are x,y,z coordinate lists describing exact distributions of tissue elements (axonal terminal fields of axons, or cell bodies) that are labeled with axonal tracing techniques. All data are translated to a common local coordinate system to facilitate across animal comparison. A search capability allows queries based on, e.g. location of tracer injection sites, tracer category, size of the injection sites, and contributing author. A graphic search tool allows the user to move a volume cursor inside a coordinate system to detect particular injection sites having connections to a specific tissue volume at chosen density levels. Tools for visualization and analysis of selected data are included, as well as an option to download individual data sets for further analysis. With this application, data and metadata from different experiments are mapped into the same information structure and made available for re-use and re-analysis in novel combinations. The application is prepared for future handling of data from other projection systems as well as other data categories. PMID- 16344145 TI - Three-dimensional chemoarchitecture of the basal forebrain: spatially specific association of cholinergic and calcium binding protein-containing neurons. AB - The basal forebrain refers to heterogeneous structures located close to the medial and ventral surfaces of the cerebral hemispheres. It contains diverse populations of neurons, including the cholinergic cortically projecting cells that show severe loss in Alzheimer's and related neurodegenerative diseases. The basal forebrain does not display any cytoarchitectural or other structural features that make it easy to demarcate functional boundaries, a problem that allowed different investigators to propose different organizational schemes. The present paper uses novel three-dimensional reconstructions and numerical analyses for studying the spatial organization of four major basal forebrain cell populations, the cholinergic, parvalbumin, calbindin and calretinin containing neurons in the rat. Our analyses suggest that the distribution of these four cell populations is not random but displays a general pattern of association. Within the cholinergic space (i.e. the volume occupied by the cortically projecting cholinergic cell bodies) the three other cell types form twisted bands along the longitudinal axis of a central dense core of cholinergic cells traversing the traditionally defined basal forebrain regions, (i.e. the medial septum, diagonal bands, the substantia innominata, pallidal regions and the bed nucleus of the stria terminalis). At a smaller scale, the different cell types within the cholinergic space occupy overlapping high-density cell clusters that are either chemically uniform or mixed. However, the cell composition of these high-density clusters is regionally specific. The proposed scheme of basal forebrain organization, using cell density or density relations as criteria, offers a new perspective on structure-function relationship, unconstrained by traditional region boundaries. PMID- 16344146 TI - No postnatal doubling of number of neurons in human Broca's areas (Brodmann areas 44 and 45)? A stereological study. AB - In this study we explored whether a postnatal doubling of the total number of neurons occurs in the human Brodmann areas 44 and 45 (Broca's area). We describe the most recent error prediction formulae and their application for the modern stereological estimators for volume and number of neurons. We estimated the number of neurons in 3D optical disector probes systematically random sampled throughout the entire Brodmann areas (BA) 44 and 45 in developing and young adult cases. In the relatively small number of male and female cases studied no substantial postnatal increase in total number of neurons occurred in areas 44 and 45; the volume of these areas reached adult values around 7 years. In addition, we did find indications that a shift from a right-over-left to a left over-right asymmetry may occur in the volume of BA 45 during postnatal development. No major asymmetry in total number of neurons in BA 44 and 45 was detected. PMID- 16344147 TI - Entorhinal projections terminate onto principal neurons and interneurons in the subiculum: a quantitative electron microscopical analysis in the rat. AB - The synaptic organization of projections to the subiculum from superficial layers of the lateral and medial entorhinal cortex was analyzed in the rat, using anterograde neuroanatomical tracing followed by electron microscopical quantification. Our aim was to assess the synaptic organization and whether the two projection components (lateral, medial) within the perforant pathway are qualitatively and quantitatively similar with respect to the types of synapses formed and with respect to the postsynaptic targets of these entorhinal projections. The tracer biotinylated dextran amine (BDA) was injected into the lateral and medial entorhinal cortex, respectively, and resulting anterograde labeling in the subiculum was studied. For each of the two projection components, we analyzed in four animals (2 x 2) a total of 100 synapses/animal with respect to features of the synapse type, i.e. asymmetrical or symmetrical, as well as regarding their postsynaptic target, i.e. dendritic shaft or spine. No clear differences were observed between the two pathways. The majority of the synapses were of the asymmetrical type, making contact with spines (78%) or with dendritic shafts (14%). A low percentage of symmetrical synapses targeted dendritic shafts (4.2%) or spines (1.3%). About 2.5% of the synapses remained undetermined. The findings indicate that the majority of entorhinal fibers reaching the subiculum exert an excitatory influence primarily onto principal neurons, with a much smaller feed forward inhibitory component. Only a small percentage of entorhinal fibers in the subiculum appears to be inhibitory, largely influencing interneurons. PMID- 16344148 TI - Morphological characteristics and electrophysiological properties of CA1 pyramidal neurons in macaque monkeys. AB - Little is known about the morphological characteristics and intrinsic electrophysiological properties of individual neurons in the nonhuman primate hippocampus. We have used intracellular recording and biocytin-labeling techniques in the in vitro hippocampal slice preparation to provide quantitative evaluation of the fundamental morphological and intrinsic electrophysiological characteristics of macaque monkey CA1 pyramidal neurons. These neurons have previously been studied in the rat in our laboratory. Monkey CA1 pyramidal neurons have an average soma volume of 3578 microm3, 4.71 basal dendrites with 53 terminal branches for a dendritic length of about 10,164 microm, 1.13 apical dendrites with 47 terminal branches for a dendritic length of about 10,678 microm. In comparison, rat CA1 pyramidal neurons have an average soma volume of 2066 microm3, 3.35 basal dendrites with 29 terminal branches for a dendritic length of about 4,586 microm, 1.43 apical dendrites with 62 terminal branches for a dendritic length of about 8,838 microm. The basic intrinsic electrophysiological properties of CA1 pyramidal cells are similar in monkeys and rats. Monkey CA1 pyramidal neurons have a resting membrane potential of about -62 mV (rat: -62 mV), an input resistance of 35 MOmega (rat: 34-49 MOmega), a rheobase of 0.17 nA (rat: 0.12-0.20 nA) and an action potential amplitude of 83 mV (rat: 71-89 mV). Although morphological differences such as the increased dendritic length may translate into differences in neural processing between primates and rodents, the functional significance of these morphological differences is not yet clear. Quantitative studies of the primate brain are critical in order to extrapolate information derived from rodent studies into better understanding of the normal and pathological function of the human hippocampus. PMID- 16344149 TI - Estimators of the precision of stereological estimates: an example based on the CA1 pyramidal cell layer of rats. AB - Because of the complex and dynamic structure of the brain, there is perhaps no other organ system in which the application of stereological methods can contribute so much with regard to understanding normal and pathological processes. In order to design the studies in an optimal manner, with regard to the number of individuals, sections, probes, and to be able to critically evaluate the stereological studies made by others, it is important for neuroscientists to have an understanding of the precision or reproducibility of a stereological estimation procedure. This precision or reproducibility is often referred to as the coefficient of error of the estimate, which is a statistical expression for the size of the standard error of the mean of repeated estimates, relative to the mean of the estimates. Like the 'margin of error' associated with public opinion polls, it indicates how much the estimate would vary if it were repeated numerous times. It is difficult and time consuming to empirically derive the coefficient of error of estimates made of features observed in histological preparations. To overcome this obstacle, it is common practice to try to get a feeling for the precision of an estimate by estimating the coefficient of error itself. In this paper, we will compare and discuss the coefficient of error of estimates of volume and cell number made with different numbers of sections and probes in the CA1 pyramidal cell layer of the rat hippocampus. The conclusions drawn from this analysis indicate that, using practically feasible and anatomically sensible sampling schemes, the Gundersen-Jensen coefficient of error estimator or the 'Split-Sample' coefficient of error estimator can provide useful information about the precision of stereological estimates even in highly irregular brain regions and requires little work. PMID- 16344150 TI - Ultrastructural quantification of glutamate receptors at excitatory synapses in hippocampus of synapsin I+II double knock-out mice. AB - Previous findings, mainly in in vitro systems, have shown that the density of vesicles and the synaptic efficacy at excitatory synapses are reduced in the absence of synapsins, despite the fact that transgenic mice lacking synapsins develop an epileptic phenotype. Here we study glutamate receptors by quantitative immunoblotting and by quantitative electron microscopic postembedding immunocytochemistry in hippocampus of perfusion fixed control wild type and double knock-out mice lacking synapsins I and II. In wild type hippocampus the densities of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subunits were higher (indicated for glutamate receptor subunit 1, highly significant for glutamate receptor subunits 2/3) in mossy fiber-to-cornu ammonis 3 pyramidal cell synapses than in the Schaffer collateral/commissural-to-cornu ammonis 1 pyramidal cell synapses, the two synapse categories that carry the main excitatory throughput of the hippocampus. The opposite was true for N-methyl-D aspartate receptors. The difference in localization of glutamate receptor subunit 1 receptor subunits was increased in the double knock-out mice while there was no change in the overall expression of the glutamate receptors in hippocampus as shown by quantitative Western blotting. The increased level of glutamate receptor subunit 1 at the mossy fiber-to-cornu ammonis 3 pyramidal cell synapse may result in alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors with reduced proportions of glutamate receptor subunit 2, and hence increased Ca2+ influx, which could cause increased excitability despite of impaired synaptic function (cf. [Krestel HE, Shimshek DR, Jensen V, Nevian T, Kim J, Geng Y, Bast T, Depaulis A, Schonig K, Schwenk F, Bujard H, Hvalby O, Sprengel R, Seeburg PH (2004) A genetic switch for epilepsy in adult mice. J Neurosci 24:10568-10578]), possibly underlying the seizure proneness in the synapsin double knock-out mice. In addition, the tendency to increased predominance of N-methyl-d-aspartate receptors at the main type of excitatory synapse onto cornu ammonis 1 pyramidal cells might contribute to the seizure susceptibility of the synapsin deficient mice. The results showed no significant changes in the proportion of 'silent' Schaffer collateral/commissural synapses lacking alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid receptors or in the synaptic membrane size, indicating that plasticity involving these parameters is not preferentially triggered due to lack of synapsins. PMID- 16344151 TI - Ionotropic glutamate receptors and glutamate transporters are involved in necrotic neuronal cell death induced by oxygen-glucose deprivation of hippocampal slice cultures. AB - Organotypic hippocampal slice cultures represent a feasible model for studies of cerebral ischemia and the role of ionotropic glutamate receptors in oxygen glucose deprivation-induced neurodegeneration. New results and a review of existing data are presented in the first part of this paper. The role of glutamate transporters, with special reference to recent results on inhibition of glutamate transporters under normal and energy-failure (ischemia-like) conditions is reviewed in the last part of the paper. The experimental work is based on hippocampal slice cultures derived from 7 day old rats and grown for about 3 weeks. In such cultures we investigated the subfield neuronal susceptibility to oxygen-glucose deprivation, the type of induced cell death and the involvement of ionotropic glutamate receptors. Hippocampal slice cultures were also used in our studies on glutamate transporters reviewed in the last part of this paper. Neurodegeneration was monitored and/or shown by cellular uptake of propidium iodide, loss of immunocytochemical staining for microtubule-associated protein 2 and staining with Fluoro-Jade B. To distinguish between necrotic vs. apoptotic neuronal cell death we used immunocytochemical staining for active caspase-3 (apoptosis indicator) and Hoechst 33342 staining of nuclear chromatin. Our experimental studies on oxygen-glucose deprivation confirmed that CA1 pyramidal cells were the most susceptible to this ischemia-like condition. Judged by propidium iodide uptake, a selective CA1 lesion, with only minor affection on CA3, occurred in cultures exposed to oxygen-glucose deprivation for 30 min. Nuclear chromatin staining by Hoechst 33342 and staining for active caspase-3 showed that oxygen-glucose deprivation induced necrotic cell death only. Addition of 10 microM of the N-methyl-D-aspartate glutamate receptor antagonist MK-801, and 20 microM of the non-N-methyl-D-aspartate glutamate receptor antagonist 2,3 dihyroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline to the culture medium confirmed that both N-methyl-D-aspartate and non-N-methyl-D-aspartate ionotropic glutamate receptors were involved in the oxygen-glucose deprivation-induced cell death. Glutamate is normally quickly removed, from the extracellular space by sodium dependent glutamate transporters. Effects of blocking the transporters by addition of the DL-threo-beta-benzyloxyaspartate are reviewed in the last part of the paper. Under normal conditions addition of DL-threo-beta-benzyloxyaspartate in concentrations of 25 microM or more to otherwise untreated hippocampal slice cultures induced neuronal cell death, which was prevented by addition of 2,3 dihyroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline and MK-801. In energy failure situations, like cerebral ischemia and oxygen-glucose deprivation, the transporters are believed to reverse and release glutamate to the extracellular space. Blockade of the transporters by a subtoxic (10 microM) dose of DL-threo beta-benzyloxyaspartate during oxygen-glucose deprivation (but not during the next 48 h after oxygen-glucose deprivation) significantly reduced the oxygen glucose deprivation-induced propidium iodide uptake, suggesting a neuroprotective inhibition of reverse transporter activity by DL-threo-beta-benzyloxyaspartate during oxygen-glucose deprivation under these conditions. Adding to this, other results from our laboratory have demonstrated that pre-treatment of the slice cultures with glial cell-line derived neurotrophic factor upregulates glutamate transporters. As a logical, but in some glial cell-line derived neurotrophic factor therapy-related conditions clearly unwanted consequence the susceptibility for oxygen-glucose deprivation-induced glutamate receptor-mediated cell death is increased after glial cell-line derived neurotrophic factor treatment. In summary, we conclude that both ionotropic glutamate receptors and glutamate transporters are involved in oxygen-glucose deprivation-induced necrotic cell death in hippocampal slice cultures, which have proven to be a feasible tool in experimental studies on this topic. PMID- 16344152 TI - Neuroprotective effects of inhibiting N-methyl-D-aspartate receptors, P2X receptors and the mitogen-activated protein kinase cascade: a quantitative analysis in organotypical hippocampal slice cultures subjected to oxygen and glucose deprivation. AB - Cell death was assessed by quantitative analysis of propidium iodide uptake in rat hippocampal slice cultures transiently exposed to oxygen and glucose deprivation, an in vitro model of brain ischemia. The hippocampal subfields CA1 and CA3, and fascia dentata were analyzed at different stages from 0 to 48 h after the insult. Cell death appeared at 3 h and increased steeply toward 12 h. Only a slight additional increase in propidium iodide uptake was seen at later intervals. The mitogen-activated protein kinases extracellular signal-regulated kinase 1 and extracellular signal-regulated kinase 2 were activated immediately after oxygen and glucose deprivation both in CA1 and in CA3/fascia dentata. Inhibition of the specific mitogen-activated protein kinase activator mitogen activated protein kinase kinase by PD98059 or U0126 offered partial protection against oxygen and glucose deprivation-induced cell damage. The non-selective P2X receptor antagonist suramin gave neuroprotection of the same magnitude as the N methyl-D-aspartate channel blocker MK-801 (approximately 70%). Neuroprotection was also observed with the P2 receptor blocker PPADS. Immunogold data indicated that hippocampal slice cultures (like intact hippocampi) express several isoforms of P2X receptors at the synaptic level, consistent with the idea that the effects of suramin and PPADS are mediated by P2X receptors. Virtually complete neuroprotection was obtained by combined blockade of N-methyl-D-aspartate receptors, P2X receptors, and mitogen-activated protein kinase kinase. Both P2X receptors and N-methyl-D-aspartate receptors mediate influx of calcium. Our results suggest that inhibition of P2X receptors has a neuroprotective potential similar to that of inhibition of N-methyl-D-aspartate receptors. In contrast, our comparative analysis shows that only partial protection can be achieved by inhibiting the extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase cascade, one of the downstream pathways activated by intracellular calcium overload. PMID- 16344153 TI - CB1 cannabinoid receptors are enriched in the perisynaptic annulus and on preterminal segments of hippocampal GABAergic axons. AB - Cannabinoids have been shown to modulate the inhibitory effect of cholecystokinin containing GABAergic interneurons in the hippocampus via type 1 cannabinoid receptors (CB1 receptor). Although immunohistochemical studies, using pre embedding techniques, have demonstrated that these receptors are abundant on GABAergic axon terminals, little is known about their exact location relative to the synapse. Here we used two recently developed antibodies against the CB1 receptor to study this question with the postembedding immunogold method, which allows the quantitative examination of receptor distribution along the axonal membrane, even within the synaptic active zone. CB1 receptor positive terminals target both the dendritic and somatic surface of neurons in the CA1 area of the rat hippocampus. We found no difference between these two populations of terminals either in their CB1 receptor density or in the distribution of receptors on their membrane. Recent studies suggest that endocannabinoids play a role in retrograde signaling at these synapses, i.e. signaling molecules diffuse from the postsynaptic membrane to nearby presynaptic terminals. Therefore, we examined the distribution of CB1 receptors on the terminal membranes. We found that they are rare in the synaptic active zone, but are enriched in the perisynaptic annulus, where they can directly influence synaptic calcium channels. Perisynaptic CB1 receptors represent about one tenth of all CB1 receptors in a terminal. In contrast, CB1 receptors have a lower density on the extrasynaptic membrane of terminals far from the postsynaptic cell. We estimated that these terminals contain exceptionally large numbers of CB1 receptors, i.e. a single axon terminal was usually labeled with more than 450 particles. An unexpected finding was that the density of CB1 receptors was significantly higher on preterminal axons than on synaptic terminals. These observations suggest that endocannabinoid signaling may subserve roles other than simply reducing transmitter release from axon terminals. PMID- 16344154 TI - Newly generated dentate granule cells from epileptic rats exhibit elongated hilar basal dendrites that align along GFAP-immunolabeled processes. AB - Previous studies showed that neurogenesis occurs in the dentate gyrus of the adult rodent. Recent evidence suggests that the resulting newly born neurons integrate into pre-existing hippocampal circuitry. Newly born neurons in the developing and adult dentate gyrus exhibit a transient basal dendrite. In adult pilocarpine-induced epileptic rats, basal dendrites persist and are ectopically located in the hilus where they receive synaptic input from mossy fiber axons. We hypothesize that these hilar basal dendrites are derived from newly born neurons that are born after the pilocarpine-induced seizures. To test this hypothesis, the length of basal dendrites from epileptic rats was compared with that from control rats using doublecortin immunocytochemistry, which labels newly born neurons and their processes for up to 3 weeks after their genesis. The data on hilar basal dendrites in pilocarpine animals indicate that those from newly born neurons are significantly longer than those found in the control rats. We also demonstrate that 20% of newly born neurons in the epileptic rat have a basal dendrite that enters the hilus at an angle greater than 30 degrees from its cell body as compared with <2% in the control rats. Lastly, we provide evidence that the hilar basal dendrites in the epileptic rats are adjacent to glial fibrillary acidic protein-labeled astrocytic processes in the hilus and suggest that an ectopic glial scaffold in the hilus is involved with the formation of hilar basal dendrites. In conclusion, the data show that newly born neurons from epileptic rats have longer hilar basal dendrites and their formation might relate to gliosis which occurs as a result of hilar neuronal cell loss after status epilepticus. PMID- 16344155 TI - Neurosteroid synthesis in the hippocampus: role in synaptic plasticity. AB - Neurosteroids are still found in the brain after steroidogenic glands were removed, indicating that they are synthesized either de novo or from endogenous precursors by enzymes present in the CNS. In fact, steroidogenic acute regulatory protein, and aromatase, two molecules essential for estrogen synthesis, are expressed in the hippocampus. We recently showed, for the first time, that estrogens are synthesized de novo in hippocampal neurons and that these hippocampus-derived estrogens are essential for synaptic plasticity. Both estrogen receptor isoforms, estrogen receptor alpha and estrogen receptor beta, are expressed in the hippocampus, and estradiol treatment of the cultures leads to an upregulation of estrogen receptor alpha. This finding confirmed the presence of functional estrogen receptors in hippocampal neurons and showed the responsiveness of the cultured hippocampal neurons to estradiol. By using letrozole, an inhibitor of aromatase, estradiol levels in hippocampal dispersion cultures as well as in hippocampal slice cultures were significantly suppressed which in turn led to a downregulation of estrogen receptor alpha. Letrozole treatment was followed by a significant decrease in the density of spines and spine synapses and in the number of presynaptic boutons. Quantitative immunohistochemistry revealed a dose-dependent downregulation of spinophilin, a spine marker, and of synaptophysin, a presynaptic marker, and of growth associated protein 43 after letrozole treatment. Our data provide strong evidence for estrogens being potent modulators of structural synaptic plasticity and point to a paracrine rather than endocrine mechanism of estrogen action in the hippocampus. PMID- 16344156 TI - Convergence of auditory-nerve fiber projections onto globular bushy cells. AB - Globular bushy cells are a key element of brainstem circuits that mediate the early stages of sound localization. Many of their physiological properties have been attributed to convergence of inputs from the auditory nerve, many of which are large with complex geometry, but the number of these terminals contacting individual cells has not been measured directly. Herein we report, using cats as the experimental model, that this number ranged greatly (9-69) across a population of 12 cells, but over one-half of the cells (seven of 12) received between 15 and 23 inputs. In addition, we provide the first measurements of cell body surface area, which also varies considerably within this population and is uncorrelated with convergence. For one cell, we were able to document axonal structure over a distance greater than 100 microm, between the soma and the location where the axon expanded to its characteristic large diameter. These data were combined with accumulated physiological information on vesicle release, receptor kinetics and voltage-gated ionic conductances, and incorporated into computational models for four cells that are representative of the structural variation within our sample population. This predictive model reveals that basic physiological features, such as precise first spike latencies and peristimulus time histogram shapes, including primary-like with notch and onset-L, can be generated in these cells without including inhibitory inputs. However, phase locking is not significantly enhanced over auditory-nerve fibers. These combined anatomical and computational approaches reveal additional parameters, such as active zone density, nerve terminal size, numbers and sources of inhibitory inputs and their activity patterns, that must be determined and incorporated into next-generation models to understand the physiology of globular bushy cells. PMID- 16344157 TI - Granule cells in the cochlear nucleus sensitive to sound activation detected by Fos protein expression. AB - Granule cells are the smallest neuronal type in the cochlear nucleus (CN). Due to their small size, it is extremely difficult to record their sound-evoked activity with microelectrodes. Compared with large, non-granule cells, much less is known about their response properties to sound stimulation. Here, we use Fos, the nuclear regulatory protein, as a neuronal activity marker to determine the responsiveness of granule cells to sound in comparison to the larger neurons. The present study determined the threshold sensitivity and activation pattern of neurons in the three subdivisions of the CN with free-field sound stimulation in monaural, awake rats. Immunocytochemical localization of Fos was used as our metric for "sound activation." Neuronal types upregulating Fos expression in response to sound stimulation were further identified with Nissl counterstaining. Our results show that most CN cell types can upregulate Fos expression when sound activated and the number of Fos-expressing neurons is directly related to sound intensity. The threshold for Fos activation in granule cells is lower than that for non-granule cells. The number of Fos activated granule cells saturates at high sound intensity, while the number of Fos activated non-granule cells is a monotonic function. By comparing the patterns of sound-induced Fos expression in different CN cell types, it may be possible to predict features of sound-evoked activity in granule cells. PMID- 16344158 TI - Laminar inputs from dorsal cochlear nucleus and ventral cochlear nucleus to the central nucleus of the inferior colliculus: two patterns of convergence. AB - The central nucleus of the inferior colliculus is a laminated structure composed of oriented dendrites and similarly oriented afferent fibers that provide a substrate for tonotopic organization. Although inputs from many sources converge in the inferior colliculus, how axons from these sources contribute to the laminar pattern has remained unclear. Here, we investigated the axons from the cochlear nuclei that terminate in the central nucleus of the cat and rat. After characterization of the best frequency of the neurons at the injection sites in the cochlear nucleus, the neurons were labeled with dextran in order to visualize their axons and synaptic boutons in the central nucleus. Quantitative methods were used to determine the size and distribution of the boutons within the laminar organization. Two components in the laminae were identified: (1) a narrow axonal lamina that included the largest fibers and largest boutons; (2) a wide axonal lamina, surrounding the narrow lamina, composed of thin fibers and only small boutons. The wide lamina was approximately 30-40% wider than the narrow lamina, and it often extended more than 100 microm beyond the larger boutons on each side. The presence of both thick and thin fibers within the acoustic striae following these injections suggests that large and small fibers/boutons within these bands may originate from different neuronal types in the dorsal and ventral cochlear nucleus. We conclude that the narrow laminae that contain large fibers and boutons originate from larger cell types in the cochlear nucleus. In contrast, the wide lamina composed exclusively of small boutons may represent an input from other, perhaps smaller neurons in the cochlear nucleus. Thus, two types of inferior colliculus laminar structures may originate from the cochlear nucleus, and the small boutons in the wide laminae may contribute a functionally distinct input to the neurons of the inferior colliculus. PMID- 16344159 TI - Relation between intrinsic connections and isofrequency contours in the inferior colliculus of the big brown bat, Eptesicus fuscus. AB - Information processing in the inferior colliculus depends on interactions between ascending pathways and intrinsic circuitry, both of which exist within a functional tonotopic organization. To determine how local projections of neurons in the inferior colliculus are related to tonotopy, we placed a small iontophoretic injection of biodextran amine at a physiologically characterized location in the inferior colliculus. We then used electrophysiological recording to place a grid of small deposits of Chicago Sky Blue throughout the same frequency range to specify an isofrequency contour. Using three-dimensional computer reconstructions, we analyzed patterns of transport relative to the physiologically determined isofrequency contour to quantify the extent of the intrinsic connection lamina in all three dimensions. We also performed a quantitative analysis of the numbers of cells in different regions relative to the biodextran amine injection. Biodextran amine-labeled fibers were mainly located dorsomedial to the injection site, confined within the isofrequency contour, but biodextran amine-labeled cells were mainly located ventrolateral to the injection site. When we counted numbers of labeled cells classified by morphological type, we found that both elongate and multipolar cells were labeled within the isofrequency contour. Because the dendrites of multipolar cells typically extend outside the isofrequency lamina, it is likely that they receive input from other isofrequency contours and relay it to more dorsomedial portions of their specific isofrequency contour, along with the frequency-specific projections of the elongate cells. Within a given isofrequency contour, there is a consistent organization in which intrinsic connections ascend from the ventrolateral portion to more dorsomedial points along the contour, forming a cascaded system of intrinsic feedforward connections that seem ideally suited to provide the delay lines necessary to produce several forms of selectivity for temporal patterns in inferior colliculus neurons. PMID- 16344160 TI - The inferior colliculus of the rat: quantitative immunocytochemical study of GABA and glycine. AB - Both GABA and glycine (Gly) containing neurons send inhibitory projections to the inferior colliculus (IC), whereas inhibitory neurons within the IC are primarily GABAergic. To date, however, a quantitative description of the topographic distribution of GABAergic neurons in the rat's IC and their GABAergic or glycinergic inputs is lacking. Accordingly, here we present detailed maps of GABAergic and glycinergic neurons and terminals in the rat's IC. Semithin serial sections of the IC were obtained and stained for GABA and Gly. Images of the tissue were digitized and used for a quantitative densitometric analysis of GABA immunostaining. The optical density, perimeter, and number of GABA- and Gly immunoreactive boutons apposed to the somata were measured. Data analysis included comparisons across IC subdivisions and across frequency regions within the central nucleus of the IC. The results show that: 1) 25% of the IC neurons are GABAergic; 2) there are more GABAergic neurons in the central nucleus of the IC than previously estimated; 3) GABAergic neurons are larger than non-GABAergic; 4) GABAergic neurons receive less GABA and glycine puncta than non-GABAergic; 5) differences across frequency regions are minor, except that the non-GABAergic neurons from high frequency regions are larger than their counterparts in low frequency regions; 6) differences within the laminae are greater along the dorsomedial-ventrolateral axis than along the rostrocaudal axis; 7) GABA and non GABAergic neurons receive different numbers of puncta in different IC subdivisions; and 8) GABAergic puncta are both apposed to the somata and in the neuropil, glycinergic puncta are mostly confined to the neuropil. PMID- 16344161 TI - Auditory thalamic organization: cellular slabs, dendritic arbors and tectothalamic axons underlying the frequency map. AB - A model of auditory thalamic organization is presented incorporating cellular laminae, oriented dendritic arbors and tectothalamic axons as a basis for the tonotopic map at this level of the central auditory system. The heart of this model is the laminar organization of neuronal somata in the ventral division of the medial geniculate body (MGV) of the rabbit, visible in routine Nissl stains. Microelectrode studies have demonstrated a step-wise ascending progression of best frequencies perpendicular to the cell layers. The dendritic arbors of MGV neurons are aligned parallel to the cellular laminae and dendritic tree width along the frequency axis corresponds closely with the frequency steps seen in microelectrode studies. In the laminated subdivision, tectothalamic axons terminate in the form of bands closely aligned with the laminae and dendritic arbors of thalamic relay neurons. The bands of tectothalamic axons extend in the anterior-posterior (A-P) plane forming a dorsal-ventral series of stacked frequency slabs. In the pars ovoidea region, the homologous spiraling of somata, dendritic fields and tectothalamic axons appear to represent a low-frequency area in this species. At least two types of tectothalamic terminals were found within the bands: large boutons frequently arranged in a glomerular pattern and smaller boutons arising from fine caliber axons. We propose that the rabbit is an ideal model to investigate the structural-functional basis of functional maps in the mammalian auditory forebrain. PMID- 16344162 TI - Quantitative assessment of developing afferent patterns in the cat inferior colliculus revealed with calbindin immunohistochemistry and tract tracing methods. AB - The central nucleus of the inferior colliculus (CNIC) is comprised of an orderly series of fibrodendritic layers. These layers include integrative circuitry for as many as 13 different ascending auditory pathways, each tonotopically ordered. Calcium-binding proteins, such as calbindin-D28k (CB), may be useful neurochemical markers for specific subsets of afferent input in these layers and their spatial organization that are developmentally regulated. In this study, CB immunohistochemistry was used to examine 1-42 postnatal-day-old kitten and adult cat CNIC and anterograde tracers were used to label afferent projections from the lateral superior olivary nucleus (LSO) to the CNIC at similar ages. A distinct axonal plexus that is CB-immunopositive is described. This CB-afferent compartment is present at birth and persists throughout the ages examined. Already at birth, the CB-immunostained plexus in kitten CNIC is organized into discrete bands that are approximately 75 microm thick and 500 microm long. In adult CNIC, the periodic banded pattern of CB-immunostained fibers is similar to that in kittens albeit bands are thicker (145 microm) and longer (700 microm). Growth in band thickness in adult cat appears proportional to growth of the IC, whereas length of the dense CB-immunostained bands is somewhat more focused in the central region of fibrodendritic layers. The banded pattern of the CB immunostained plexus is well correlated with the location and dimension of afferent projections from the LSO in newborn kitten labeled with carbocyanine dye, 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate and in adult cat labeled with wheat germ agglutinin conjugated with horseradish peroxidase. The results reveal a neurochemical marker for one type of synaptic compartment in CNIC layers, banding, that is organized before hearing onset in kittens, but that may undergo some postnatal pruning. PMID- 16344163 TI - Unilateral cochlear ablation in adult ferrets results in upregulation in calretinin immunostaining in the central nucleus of the inferior colliculus. AB - In the present study, unilateral cochlear ablations were performed in adult ferrets in order to determine whether an upregulation of the calretinin immunostained plexus in the central nucleus of the inferior colliculus occurs and if so, what the time course of this upregulation is. Accordingly, the mean gray level and the calretinin-immunostained area of the axonal plexus in the central nucleus of the inferior colliculus were evaluated at 1, 20 and 90 days after cochlear ablation. In unoperated animals, the calretinin-immunostained plexus was bilaterally symmetric. In ablated animals, both the mean gray level and the immunostained area of the plexus increased in the central nucleus of the inferior colliculus contralateral to the lesion compared with both the ipsilateral side and unoperated animals. This upregulation was present 24 h after the ablation and did not change at the two subsequent time points. In a previous study in young ferrets, the immunostained area of the plexus in the central nucleus of the inferior colliculus contralateral to the lesion increased 200% compared with control ferrets [J Comp Neurol 460 (2003) 585], whereas it increased only 33% in adult ferrets. These findings suggest that 1) calretinin upregulation in the contralateral central nucleus of the inferior colliculus following cochlear ablation occurs by 24 h after cochlear ablation and 2) there is an age-related decline in the magnitude of this upregulation after cochlear ablation. PMID- 16344165 TI - A historical report on Robert Kienbock (1871-1953) and Kienbock's Disease. AB - Robert Kienbock was the first to identify and describe Kienbock's disease. Considered one of the early fathers of radiology, he was a prolific researcher who made an immense contribution to the early development of radiology. In addition to groundbreaking work in therapeutic radiology Kienbock had a special interest in diseases of the bones and joints, especially those that involved the hand. Kienbock's contribution to medicine, particularly the diagnosis of hand disorders, continues to resonate today. PMID- 16344166 TI - Complications and outcome of four-corner arthrodesis: circular plate fixation versus traditional techniques. AB - PURPOSE: To review the clinical and radiographic results of scaphoid excision and four-corner arthrodesis using a circular plate and screws compared with traditional fusion techniques (wires, staples, screws). METHODS: Fifty-eight patients with four-corner arthrodesis (plate fixation, n = 27; traditional fixation, n = 31) were evaluated for radiographic and clinical success using wrist radiographs and functional assays. Patients were subjectively surveyed using the standardized Disabilities of the Arm, Shoulder, and Hand questionnaire and classification scales for pain and satisfaction. Objective measurements included grip-strength and range-of-motion measurements. RESULTS: Radiographic analysis showed 26% nonunion with loose hardware in the plate group compared with 3% in the traditional group and 22% hardware impingement in the plate group compared with 3% in the traditional group. Clinical evaluation yielded a mean grip strength of 31 kg (70% of opposite side) for plate fixation and 33 kg (79% of opposite side) for traditional fixation. The mean flexion-extension arc was 48% and 50% of the opposite wrist for plate and traditional patients, respectively. The mean adjusted Disabilities of the Arm, Shoulder, and Hand questionnaire scores were 27 out of 100 for plate patients and 8 out of 100 for traditional patients. Pain classification scores showed that only 2 patients in the plate group were pain free whereas there were 8 patients in the traditional group who were pain free. Overall patient satisfaction was 60% for the plate group whereas the traditional patient group reported 100% satisfaction. CONCLUSIONS: The rate of major complications (nonunion or impingement) was much greater with circular plate fixation (48%) versus traditional fixation techniques (6%). With the plate procedure the grip strength and arc of motion decreased approximately 30% and 52%, respectively, compared with decreases of 21% and 50%, respectively, for traditional fusion methods. Additionally, subjective patient dissatisfaction was 40% in the plate group compared with 0% in the traditional group. We postulate that the increased complication and dissatisfaction rates associated with plate fixation may be attributable to possible biomechanical imperfections or increased technical demands with this fusion system. PMID- 16344167 TI - Canine carpal joint fusion: a model for four-corner arthrodesis using a porous tantalum implant. AB - PURPOSE: Interest has focused on porous materials that promote bony ingrowth. In this study a porous tantalum implant was used as an adjunct to intercarpal stabilization in a canine model of wrist arthrodesis. METHODS: A defect was created at the junction of the radiocarpal, ulnocarpal, and fourth carpal bones, analogous to a four-corner fusion site in humans. A tantalum cylinder was press fit and stabilized with K-wires. Controls were represented by creating the defect without implant placement. Animals were killed at 4, 8, and 12 weeks. RESULTS: Histology showed bony ingrowth as early as 4 weeks and mechanical testing showed a statistically significant increase in strength of the construct over time. Controls failed to achieve union at any time point. CONCLUSIONS: The implant served as an adjunct to stabilization of the carpus in this model of four-corner fusion, suggesting a novel application of this material in conditions in which bone graft has been required previously. This study represents a preliminary investigation of the use of a tantalum device for intercarpal stabilization; it does not compare this technique with conventional methods. PMID- 16344168 TI - Patterns of carpal deformity in scaphoid nonunion: a 3-dimensional and quantitative analysis. AB - PURPOSE: To clarify quantitatively the 3-dimensional deformity of the carpus in scaphoid nonunion on the basis of fracture location. METHODS: Three-dimensional computed tomography was used to examine 20 patients with scaphoid nonunion. Displacements of the distal and proximal fragments of the scaphoid, lunate, triquetrum, and capitate were visualized and quantified using a 3-dimensional image-matching technology. Cases were categorized as distal fracture (16 cases) or proximal fracture (4 cases) based on the location of the fracture line relative to the dorsal apex of the scaphoid ridge where the dorsal scapholunate interosseous ligament is attached. RESULTS: The displayed distal scaphoid fractures showed that the proximal fragment of the scaphoid, lunate, and triquetrum rotated into extension and supination. The distal fragment of the scaphoid and capitate translated dorsally without notable rotation. The deformity in proximal fractures was less remarkable than that in distal fractures. Most distal scaphoid nonunions had dorsal intercalated segment instability deformity patterns, whereas a dorsal intercalated segment instability occurred in only 1 case of a proximal fracture. CONCLUSIONS: Whether the fracture line passes distal or proximal to the dorsal apex of the scaphoid determines the subsequent carpal deformity. Dorsal translation of the distal fragment might be one of the factors in the development of degenerative change at the radial styloid. PMID- 16344169 TI - Trans-scaphoid perilunate fracture dislocations: results of screw fixation of the scaphoid and lunotriquetral repair with a dorsal approach. AB - PURPOSE: To review the clinical and radiographic outcome of dorsal trans-scaphoid perilunate fracture-dislocations treated with screw fixation of the scaphoid and repair of the lunotriquetral ligament with bone anchors. METHODS: Twenty-five patients treated over an 11-year period for dorsal trans-scaphoid perilunate fracture-dislocations were reviewed retrospectively at a mean of 44.3 months. The mean age of the patients was 28.6 years, and 22 patients were men. Delay to surgery was 3.5 days. Five scaphoids had bone grafting from the distal radius. Six patients developed carpal tunnel syndrome requiring release. The functional outcome was determined by comparing the range of motion of the injured extremity with the uninjured extremity, grip strength, ability to return to pre-injury employment, and overall patient satisfaction. Radiographic evaluation comprised time to scaphoid union, any changes in the lunotriquetral interval, development of a volar intercalated segmental instability pattern, and any development of arthritis over time. RESULTS: Total range of motion achieved was 91% of the uninjured wrist and grip strength was 80%. Average extension was 54 degrees with an average flexion of 60 degrees . Ulnar deviation was 23 degrees with radial deviation averaging 18 degrees . The average supination was 76 degrees and the average pronation was 76 degrees . All scaphoids united primarily. The average time to union of the scaphoid was 16 weeks. For those scaphoids with bone grafting the union time was an average of 18.4 weeks. The average postreduction lunotriquetral gap was 1.8 +/- 0.4 mm. The average lunotriquetral gap at the last follow-up evaluation was 1.9 +/- 0.6 mm. None of the patients developed a volar intercalated segmental instability deformity. All but 2 patients returned to their pre-injury occupation. All patients, however, returned to some type of employment. Complications included 1 superficial pin track infection that resolved with removal of the pin and a short course of oral antibiotics. CONCLUSIONS: A dorsal approach to the wrist provides adequate exposure for reduction of carpal bones, internal fixation of the scaphoid, and lunotriquetral repair. Although perilunate fracture-dislocations are challenging problems to treat, all of the patients had acceptable pain relief and achieved sufficient range of motion and strength to return to gainful employment. PMID- 16344170 TI - Simultaneous fractures of the capitate and hamate in the coronal plane: case report. AB - Fractures of the carpal bones in the coronal plane are encountered rarely and their diagnosis usually is delayed. Coronal hamate fractures are relatively well described but a coronal capitate fracture occurring simultaneously with a coronal hamate fracture is extremely rare. We describe such an injury addressed with fixation of the hamate alone. PMID- 16344172 TI - A radiographic view of the scaphotrapezium-trapezoid joint. AB - Traditionally the scaphotrapezium-trapezoid joint is imaged through a posteroanterior view of the wrist. We describe an x-ray view that is aimed directly at the joint, which gives better visualization than the standard views. PMID- 16344171 TI - Changes in patterns of scaphoid and lunate motion during functional arcs of wrist motion induced by ligament division. AB - PURPOSE: To determine the in vitro motion of the scaphoid and lunate during wrist circumduction and wrist dart-throw motions and to see how these motions change after the ligamentous stabilizers of the scaphoid and lunate are sectioned in a manner simulating scapholunate instability. METHODS: Twenty-one fresh-frozen cadaver forearms were moved through a dart-throw motion and a circumduction motion using a wrist joint simulator. Scaphoid and lunate motion were measured with the wrist ligaments intact and after sectioning of the scapholunate interosseous ligament, the scaphotrapezium ligament, and the radioscaphocapitate ligament. RESULTS: In the intact wrist the scaphoid and lunate moved more during circumduction than during the dart-throw motion. With ligamentous sectioning the scaphoid flexed more and the lunate extended more during both the circumduction and dart-throw motions. During the circumduction motion both before and after sectioning the global motion of the scaphoid was greater than that of the lunate. After sectioning the scaphoid motion increased and the lunate motion decreased. CONCLUSIONS: The scaphoid and lunate motions were observed to change remarkably after ligamentous sectioning. The observed changes in carpal motion correlate with the clinical observation that after ligamentous injury arthritic changes occur in the radioscaphoid joint and not in the radiolunate joint. Analysis of the injured wrist in positions that combine flexion-extension and radial-ulnar deviation may allow noninvasive diagnosis of specific wrist ligament injuries. PMID- 16344173 TI - Contribution of the interosseous membrane to distal radioulnar joint constraint. AB - PURPOSE: Although forearm injuries are accompanied frequently by rupture to the interosseous membrane (IOM) diagnosis of the extent of IOM injury is difficult. In this study we evaluated distal radioulnar joint (DRUJ) laxity caused by both partial and complete IOM disruption and compared these quantitative measurements with the common clinical manual evaluation of DRUJ laxity and dislocatability. METHODS: Human cadaveric forearms (n = 8) were used in this study. Skin, muscles, and tendons were removed. The specimens were mounted on an experimental apparatus that allowed the radius to move freely about the fixed ulna. Tests were performed in neutral rotation, 60 degrees pronation, and 60 degrees supination. Under various conditions of IOM sectioning testing was performed by volary and dorsally translating the radius relative to the ulna in the coronal plane of the radius. Testing was performed both qualitatively as would be performed in the clinic and quantitatively with an instrumented probe. RESULTS: Our results show that dorsal dislocation of the radius relative to the ulna strongly suggests distal IOM rupture. Disengagement of the radius from the DRUJ indicated injury to the distal and middle IOM. The distal IOM constrained volar and dorsal laxity of the radius at the DRUJ in all forearm rotation positions. The midportion of the IOM constrained laxity except in the volar direction of the pronated forearm. The proximal IOM did not constrain the proximal radius except dorsally for the pronated forearm position. CONCLUSIONS: The IOM, in particular the distal IOM, plays an important role in constraining dorsal dislocation of the radius at the DRUJ. PMID- 16344174 TI - Ulnar-positive variance as a predictor of distal radioulnar joint ligament disruption. AB - PURPOSE: Previous cadaveric data show that disruption of the triangular fibrocartilage complex (TFCC) at the wrist allows 0.5 to 3.0 mm of proximal radius migration. Anatomic studies have documented the presence of superficial and deep fibers of both the palmar and the dorsal distal radioulnar joint (DRUJ) ligaments. The aim of this study was to determine the contribution of the superficial and deep fibers of the DRUJ ligaments to longitudinal forearm stability as measured by ulnar-positive variance. METHODS: Eight fresh-frozen cadaver specimens were included in this study. Each specimen was secured with external fixation clamps to a sequential loading frame with the elbow in 90 degrees of flexion and the forearm and wrist in neutral pronation supination, neutral ulnar-radial deviation, and neutral volar-dorsal angulation. The radial head was resected and a force gauge was applied to the proximal radius. The peripheral TFCC was identified through an incision between the fifth and sixth extensor compartments and the dorsal capsulotomy of the DRUJ capsule. After baseline measurement sequential transection of the superficial and deep fibers of the TFCC was performed. Before and after each step load application and removal were performed by attaching an 88.90-N weight to the end of a force gauge and via longitudinal traction on the proximal part of the radius, and ulnar variance was measured with wrist fluoroscopy. RESULTS: Transection of the superficial TFCC fibers resulted in radius migration of 0.70 mm. This migration, however, was not significantly different from that observed at baseline. After both the superficial and deep TFCC fibers were transected the radius migrated proximately with load. This change of ulnar variance was significantly greater than that observed at baseline or after transection of only the superficial TFCC fibers. CONCLUSIONS: Traumatic injury to the TFCC with radiographic evidence of ulnar positive variance may be an indication of disruption of the deep TFCC fibers. PMID- 16344175 TI - Influence of distal radioulnar joint subluxation on restricted forearm rotation after distal radius fracture. AB - PURPOSE: To analyze the influence of subluxation of the distal radioulnar joint (DRUJ) on restricted forearm rotation after distal radius fracture. METHODS: Twenty-two cases of healed unilateral distal radial fracture with restricted forearm rotation were included in the study. The subluxation of the DRUJ was evaluated using helical computed tomography scan at neutral, maximum pronation, and maximum supination and presented as the percent displacement of the ulnar head in both the injured and uninjured sides. The radiographic parameters of palmar tilt, radial inclination, dorsal shift, radial shift, and ulnar variance were measured on plain x-ray films and the rotational deformity of the distal radius was evaluated from the computed tomography scan. The differences of each radiographic parameter from the uninjured side were calculated. The relationships between the restricted forearm rotation and the percent displacement of the ulnar head and each of the radiographic parameters were analyzed statistically. RESULTS: When forearm pronation was restricted the ulnar head was located palmarly at neutral, maximum supination, and maximum pronation with severe dorsal tilt of the distal radius. When supination was restricted the ulnar head was located dorsally at maximum supination with severe ulnar-positive variance. CONCLUSIONS: The subluxation of the DRUJ was related to restricted forearm rotation. The radiographic parameters of palmar tilt and ulnar variance showed an adverse influence on the position of the ulnar head at the DRUJ, which might lead to restricted forearm rotation after distal radial fracture. PMID- 16344176 TI - A meta-analysis of outcomes of external fixation versus plate osteosynthesis for unstable distal radius fractures. AB - PURPOSE: External fixation and open reduction and internal fixation have been the traditional techniques for surgical fixation of unstable distal radius fractures. The existing literature has not identified which is superior, primarily because of the lack of comparative trials. We performed a comprehensive systematic review and meta-analysis of the current literature on external fixation and internal fixation of distal radius fractures to determine the dominant strategy based on available scientific evidence. METHODS: We searched MEDLINE and EMBASE for English-language articles published between 1980 and 2004 that satisfied predetermined inclusion and exclusion criteria. The outcomes of internal and external fixation were compared using continuous measures of grip strength, wrist range of motion, and radiographic alignment and categoric measures of pain, physician-rated outcome scales, and complication rates. Outcomes were pooled by random-effects meta-analysis and meta-regression analysis was used to control for patient age, presence of intra-articular fracture, duration of follow-up period, and date of publication. Sensitivity analyses were used to test the stability of the meta-analysis results under different assumptions. RESULTS: Forty-six articles were included in the review with 28 (917 patients) external fixation studies and 18 (603 patients) internal fixation studies. Meta-analysis did not detect clinically or statistically significant differences in pooled grip strength, wrist range of motion, radiographic alignment, pain, and physician rated outcomes between the 2 treatment arms. There were higher rates of infection, hardware failure, and neuritis with external fixation and higher rates of tendon complications and early hardware removal with internal fixation. Considerable heterogeneity was present in all studies and adversely affected the precision of the meta-analysis. CONCLUSIONS: The current literature offers no evidence to support the use of internal fixation over external fixation for unstable distal radius fractures. Comparative trials using appropriately sensitive and validated outcome measurements are needed to guide treatment decisions. PMID- 16344177 TI - Reconstruction of the distal radius facet by a free vascularized osteochondral autograft: anatomic study and report of a patient. AB - PURPOSE: Large chondral defects of the distal radius after fractures present a reconstructive challenge. The purpose of this study was to present the anatomic findings from a cadaver of a vascularized osteochondral autograft taken from the third metatarsal appropriate for reconstructing the distal radius articular facet. A patient is presented in whom 70% of the scaphoid fossa was reconstructed with this technique. METHODS: The base of the third metatarsal was studied in the feet of 20 cadavers. The size and shape of the cartilage were measured. Additionally vessel distribution was recorded and the diameters of vascular foramina were measured with Juch's method. RESULTS: The base of the third metatarsal is pear shaped and is wider dorsally than plantarly. It averages 19.2 mm long on its main axis. Its cartilaginous surface is minimally concave or flat and it is slanted slightly proximal-dorsal to distal-plantar and proximal peroneal to distal-tibial. Nutrient foramina were found in every case in the dorsum and on both sides of the proximal shaft. At least 1 nutrient vessel could be tracked back to the dorsalis pedis in every dissected specimen. CONCLUSIONS: The anatomic features of the base of the third metatarsal make it a potential vascularized autograft to consider for osteochondral defects of the distal radius. PMID- 16344178 TI - Madelung's deformity: quantitative assessment of x-ray deformity. AB - PURPOSE: To evaluate 5 defined measurement techniques that are applicable to x rays of Madelung's deformity: ulnar tilt, lunate subsidence, lunate fossa angle, palmar tilt, and palmar carpal displacement. The measurements rely on the longitudinal axis of the ulna and the carpal bones to determine drawing lines and avoid the distorted distal radius and its deformed lunate fossa. The reliability and reproducibility of the measurements is determined. METHODS: Forty-eight sets of posteroanterior and lateral x-ray views of the wrist of subjects with the clinical diagnosis of Madelung's deformity were measured by 4 raters. Each rater made the 5 defined measurements on each pair of x-rays. Pairs of raters were compared for reliability using the Pearson correlation coefficient and Lin's concordance correlation coefficient. Two raters repeated the 4 reliable measurements a minimum of 6 months after the first measurements. Each rater's results were compared for reproducibility using Lin's concordance correlation coefficient. RESULTS: Ulnar tilt and lunate subsidence have excellent reliability and reproducibility. Palmar carpal displacement has acceptable reliability and reproducibility. Lunate fossa angle has borderline reliability but excellent reproducibility. Palmar tilt has poor reliability. CONCLUSIONS: Ulnar tilt, lunate subsidence, and palmar carpal displacement, as defined, are considered reliable and reproducible measurements for quantifying the severity of Madelung's deformity on x-rays. Lunate fossa angle is not sufficiently reliable for comparing preoperative and postoperative wrists but may prove useful in establishing an early diagnosis. Palmar tilt is not measured reliably on a lateral x-ray because of the superimposition of multiple structures on a lateral x-ray and the absence of the volar part of the lunate fossa in patients with severe Madelung's deformity. Advanced imaging techniques are needed to delineate the deformity of the distal radius in a lateral projection. PMID- 16344179 TI - Wrist denervation and the anterior interosseous nerve: anatomic considerations. AB - PURPOSE: Wrist denervation via resection of the distal anterior interosseous nerve (AIN) and the posterior interosseous nerve (PIN) is an effective treatment for chronic wrist pain. When performing this procedure through a dorsal approach we have been impressed by anatomic variations of the AIN. This has raised concerns about potential denervation of the pronator quadratus (PQ). The purpose of this study was to elucidate the anatomy of the AIN and PIN as encountered through a dorsal distal forearm incision. METHODS: Ten fresh-frozen cadavers were dissected. Before dissection radiographs were obtained to ensure accurate localization of the proximal ulnar head with a radiopaque marker. A dorsal approach to the distal forearm was made to identify the anatomy of the PIN and AIN. The location and diameter of all AIN branches were noted by using an operating stereoscopic microscope at x 25 magnification and a precision caliper. The PIN anatomy and size also were noted. RESULTS: The anatomy of the AIN was variable. The average AIN diameter proximal to the PQ was 1.5 mm. The average number of AIN motor branches was 4.2. The largest PQ motor branch was the first motor branch and was located at an average distance of 37.9 mm from the proximal ulnar head. The last motor branch was found an average of 23.9 mm from the proximal ulnar head. In 9 of 10 specimens the sensory branch tunneled radially through the distal PQ and innervated the periosteum of the volar distal radius. In 4 of 10 specimens a separate branch to the distal radioulnar joint was present. We found an average PIN diameter of 0.87 mm. CONCLUSIONS: Resection of the AIN at a point 4 cm proximal to the proximal point of the ulnar head would denervate completely the PQ in our cadaver population. Division of the AIN 2 cm proximal to the ulnar head would spare most of the PQ motor branches. PMID- 16344180 TI - Anatomy of the lateral antebrachial cutaneous and superficial radial nerves in the forearm: a cadaveric and clinical study. AB - PURPOSE: To define the anatomy of the lateral antebrachial cutaneous nerve (LACN) and the superficial radial nerve (SRN) in relation to easily identifiable landmarks in the dorsoradial forearm to minimize the risk to both nerves during surgical approaches to the dorsal radius. METHODS: In this study 37 cadaveric forearms and 20 patients having distal radius external fixation were dissected to identify these nerves in relation to various anatomic landmarks. RESULTS: Based on these dissections the anatomy was divided into 2 zones that can be identified by easily visible and palpable landmarks. Zone 1 extends from the elbow to the cross-over of the abductor pollicis longus with the extensor carpi radialis brevis and longus. Zone 2 is distal to the cross-over. In zone 1 the 2 nerves can be differentiated through limited incisions based on their depth and anatomic location. Within this zone the SRN is deep to the brachioradialis until 1.8 cm proximal to zone 2 (9 cm proximal to the radial styloid), where it becomes superficial and pierces the fascia of the mobile wad and then remains deep to the subcutaneous fat. In contrast the LACN pierces the fascia between the brachialis and biceps muscles at the level of the elbow. In all specimens the LACN ran parallel to the cephalic vein within the subcutaneous fat. In 31 specimens it ran volar to the vein and in 5 specimens the nerve crossed under the cephalic vein at the elbow and ran dorsal to the vein in the forearm. One specimen had 2 branches with 1 on either side of the vein. Differentiation of these nerves was found to be possible through limited incisions in zone 1 during placement of external fixation pins for distal radius fractures. The LACN always was located in the superficial fat running with the cephalic vein, whereas the SRN was deeper to this nerve either covered by the brachioradialis or closely adherent to it within the investing fascia of the mobile wad. In zone 2 the nerves arborized and ran in the same tissue plane, making differentiation through limited incisions difficult. CONCLUSIONS: Dividing forearm anatomy into zones aids in understanding the complex 3-dimensional anatomy. Recognition of the consistent location of both the LACN and SRN facilitates surgical exposure. This allows localization through limited incisions during nerve repair and hardware placement, thereby enhancing uncomplicated and favorable outcomes. PMID- 16344181 TI - A variant of Kaplan's accessory branch of the dorsal cutaneous branch of the ulnar nerve: a case report and review of the literature. AB - Kaplan's accessory branch is an aberrant branch of the dorsal cutaneous branch of the ulnar nerve that arises proximal to the styloid process of the ulna and courses ulnar to the pisiform. Variations of this anomaly have been described as having an end point of connection to the sensory branch of the ulnar nerve, to the motor branch of the ulnar nerve (rare), to the digital nerve at the level of the midhypothenar eminence, or to the proximal interphalangeal joint of the small finger or as running as an independent branch to the volar aspect of the small finger. We report a variant of Kaplan's accessory branch that coursed through the insertion of the flexor carpi ulnaris, a groove on the ulnar aspect of the pisiform, and connected to the ulnar nerve trunk proximal to its bifurcation into its motor and sensory branch. Based on the findings of the case presented and a review of the literature we offer a classification system for this anomaly. PMID- 16344182 TI - The prevalence of hereditary neuropathy with liability to pressure palsies in patients with multiple surgically treated entrapment neuropathies. AB - PURPOSE: Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal-dominant peripheral neuropathy that results from deletion of a 1.5 Megabase pair (Mb) segment of the short arm (p) of chromosome 17. Hereditary neuropathy with liability to pressure palsies increases susceptibility of peripheral nerves to pressure and trauma and can be associated with symptoms at multiple anatomic entrapment sites. Many patients present with multiple upper extremity entrapment neuropathies and the etiology is uncertain. We hypothesized that some of these patients have an underlying hereditary neuropathy. The purpose of this study was to determine the prevalence of HNPP in patients with multiple surgically treated upper-extremity entrapment neuropathies. METHODS: The inclusion criterion for the study was history of more than 1 carpal tunnel release and/or ulnar nerve transposition. The exclusion criteria were history of diabetes or history of Charcot-Marie-Tooth neuropathy. Fifty-nine patients were in the study group. Two patients known to have the 17p11.2 deletion were used as controls. Genomic DNA was extracted from peripheral blood. Each sample was genotyped using polymerase chain reaction (PCR) amplification with short tandem repeat polymorphism markers within the 1.5-Mb region of 17p deleted in HNPP. Markers were scored as homozygous or heterozygous after resolution by polyacrylamide gel electrophoresis and silver staining. RESULTS: The 2 control patients were homozygous for 11 markers. None of the 59 study patients were homozygous for all markers tested in the deleted region. No study patient had the 17p deletion diagnostic for HNPP. Based on the sample size of 59 patients the 95% confidence interval for the prevalence of the 17p11.2 deletion in this population is 0% to 5%. CONCLUSIONS: We found no evidence for an association between HNPP and patients who have multiple surgical releases for upper-extremity entrapment neuropathies. PMID- 16344183 TI - Anatomic variations of the hook of hamate and the relationship to carpal tunnel syndrome. AB - PURPOSE: To determine the incidence of anatomic variations of the hook of hamate and to evaluate its association with the development of carpal tunnel syndrome (CTS). METHODS: Radiographs of 3,218 hands (2,070 patients) were evaluated. This group included 2,866 hands diagnosed with CTS and 352 hands without the diagnosis of CTS. Using the carpal tunnel view variations of the hook of hamate were identified. Variants were classified based on appearance and measurements. Patients having variant hooks of hamate who were diagnosed with CTS then were compared with patients with variant hooks of hamate who had no evidence of CTS. Endoscopic carpal tunnel release using the Chow technique was performed on all patients with CTS who had a known variation of the hook of hamate without difficulty or complication. RESULTS: Variations of the hook of hamate were found in 96 hands: 42, bipartite hook; 50, hypoplastic hook; and 4, aplastic hook. Ninety-three hands with variant hooks of hamate were diagnosed with CTS. The remaining 3 hands identified with variants had no evidence of CTS. After comparing the 2 groups, we found that the incidence of a variant hook of hamate in the group with CTS was significantly greater than the incidence of variations in the group without CTS. CONCLUSIONS: Three variations of the hook of hamate were identified with radiographic evaluation using the carpal tunnel view. Variations are more prevalent than once thought. There was a significant increase in the incidence of variation in the hook of hamate in the group with CTS compared with the group without CTS. PMID- 16344184 TI - Investigation of resistance of digital subcutaneous edema to gliding of the flexor tendon: an in vitro study. AB - PURPOSE: Although edema generally is considered to contribute to resistance to tendon mobilization and is a cause of tendon overload during postoperative motion exercise, it is unclear exactly how edema of the peritendinous tissues affects tendon mobilization. We assessed the effects of simulated subcutaneous edema on the gliding resistance of the flexor tendon in an in vitro model using chicken toes. METHODS: Thirty long toes of white Leghorn chickens were used. Twenty-two toes were divided into 2 groups and another 8 toes were used to determine the preconditions needed. In group 1 we produced 3 levels of edema severity in the subcutaneous tissue over a 1-cm segment by means of saline injection. In group 2 we created moderate tissue edema over 1-cm, 2-cm, or 3-cm segments of the toes. The work required to move the flexor digitorum profundus tendon over a fixed excursion and ultimate force were recorded with a testing machine after each run of flexion. RESULTS: Work and ultimate force increased significantly in the toes with simulated tissue edema in proportion to the severity and area of the edema. Even the least severe edema increased the work and force; a further increase to moderate edema tripled the energy needed to flex the toes. Increases in edema from 1-cm to a 2-cm and then to a 3-cm toe segment increased the work and force for each increment of extension. CONCLUSIONS: Simulated edema significantly increases energy and force required to move the tendons. The increase in resistance was proportionate to the severity and area of the edema. These results suggest that postoperative edema may increase significantly the resistance to tendon motion and that limiting both the severity and size of edema likely will reduce the resistance. PMID- 16344185 TI - Tendon healing in vitro: bFGF gene transfer to tenocytes by adeno-associated viral vectors promotes expression of collagen genes. AB - PURPOSE: Adeno-associated virus-mediated gene transfer is promising in the delivery of genes to tendons because this vector stimulates few adverse tissue reactions. Basic fibroblast growth factor (bFGF) promotes collagen production in healing tendons. We transferred the exogenous bFGF gene to proliferating tenocytes by adeno-associated viral (AAV) vectors and investigated its effects on the expression of the collagen genes in an in vitro tenocyte model. METHODS: AAV2 vectors harboring the rat bFGF gene were constructed. Tenocytes were obtained from explant cultures of rat intrasynovial tendons and were distributed into 21 culture dishes and 8 wells. Tenocytes in 7 dishes were treated with AAV2 bFGF for 3 hours and then were cultured for 10 days. Tenocytes in 14 dishes (sham vector and nontreatment controls) did not receive the transgene. Efficiency of the gene transfer was evaluated by in situ beta-galactosidase staining in 8 wells after treatment with AAV2 lacZ. Expression of the target genes was assessed by reverse transcription polymerase chain reactions with primers specifically amplifying the target genes. Expression of bFGF and type I and III collagen genes was determined by quantitative analysis of the polymerase chain reaction products. RESULTS: Positive beta-galactosidase staining confirmed the effectiveness of AAV2-mediated gene delivery to tenocytes. The level of expression of the bFGF gene was increased significantly after gene transfer. Levels of expression of type I and III collagen genes after transfer of the exogenous bFGF gene were increased significantly compared with those in the cells treated with sham vectors or in nontreatment controls. CONCLUSIONS: Delivery of exogenous bFGF gene to tenocytes can increase significantly the levels of expression of the bFGF and type I and III collagen genes. AAV2 vectors provide a novel method for delivering growth factor genes to tenocytes. These findings warrant future in vivo study of the delivery of genes pertinent to tendon healing through AAV2-based gene therapy to enhance repairs of injured flexor tendons. PMID- 16344186 TI - Core suture purchase affects strength of tendon repairs. AB - PURPOSE: It generally is considered that a certain distance should be maintained between the site of the tendon-suture junction and the laceration level of the tendon. In this study we assessed how the length of core suture purchase may affect the repair strength of transversely cut tendons using a 2-strand modified Kessler method and a 4-strand circle-locking method. METHODS: Seventy-four fresh pig flexor tendons were transected. Fifty-eight tendons were divided into 4 groups and repaired with a 2-strand grasping repair technique with the core suture purchase in the tendon stump ranging from 0.4 to 1.2 cm. Sixteen tendons were repaired with a 4-strand circle-locking tendon-suture repair technique. The core suture purchase of these tendons was 0.4 and 1.0 cm, respectively. The tendons were subjected to a linear, noncyclic, load-to-failure test in a tensile testing machine. The forces measured for initial gap formation, 2-mm gap formation, and ultimate strength were recorded for each repair. RESULTS: The resistance to gap formation and ultimate strength of 2-strand grasping technique repairs increased significantly as the suture purchase increased from 0.4 to 0.7, 1.0, and 1.2 cm although strength remained constant from 0.7 to 1.2 cm. The strength of 4-strand circle-locking repairs with a suture purchase of 1.0 cm was statistically greater than that of the repairs with a suture purchase of 0.4 cm. CONCLUSIONS: For both the 2-strand grasping and 4-strand circle-locking repair methods, the length of core suture purchase significantly influences the resistance to 2-mm gap formation and the ultimate strength of repairs in transversely lacerated flexor tendons. We determined that the optimal length of purchase is between 0.7 and 1.0 cm and that increased length of purchase from 0.7 to 1.2 cm does not increase the strength of the repair. Core suture purchase length of 0.4 cm or less results in significantly weaker repairs. PMID- 16344187 TI - Analysis of intrinsic-extrinsic muscle function through interactive 3-dimensional kinematic simulation and cadaver studies. AB - PURPOSE: To revisit the analysis of intrinsic muscle function and its relationship to the overall flexion moment at the index metacarpophalangeal (MCP) joint. Prior studies reported inconsistent levels of intrinsic contribution varying from 12.5% to 73% of the total flexion moment at the index MCP joint. This study hypothesized that 26% is a more realistic figure using computer simulation moment arms combined with known muscle-tension fractions. This study tested the hypothesis using a 2-part fresh cadaver study, the objectives of which were to measure actual muscle moment arms throughout index MCP range of motion and then combine these measurements with tension fractions to calculate normal intrinsic minus and low ulnar nerve palsy moments and then using the same specimens to apply loads to the muscles and measure resultant moments directly. METHODS: An interactive computer simulation was constructed to replicate the position and loading orientation of an earlier experiment reported in the literature and known muscle-tension fractions were combined with the resultant moment arm structure to calculate the moment at the index MCP joint for maximum isometric contraction of the normal, intrinsic minus, and low ulnar nerve palsy finger. Eleven fresh cadaver specimens were used. For experiment A the excursion and angle data were used to determine moment arms of each muscle and moments were calculated. For experiment B the muscles were connected to weights based on muscle-tension fractions and the resultant moment was measured at the fingertip. RESULTS: Experiment A determined the muscle-tendon moment arms at the index MCP joint throughout the flexion-extension range of motion. Combining the flexor moment arms with known tension fractions determined that the intrinsics contributed 24% to the total flexion moment, the dorsal and palmar interossei contributed 22%, and the lumbrical contributed 2%. The results of experiment B confirmed the results of experiment A. CONCLUSIONS: Intrinsic muscle contribution to flexion moment at the index MCP joint is not as high or as low as previously reported. The relative moment production is governed by both the moment arms and tension produced by the muscle; thus a more precise knowledge of moment arms is essential to understand muscle balance at a joint. Clinically the hand surgeon should expect a loss of 24% of index MCP flexion moment-generating capacity in the intrinsic-minus index finger and a loss of 22% in the index finger on a hand with low ulnar nerve palsy. Furthermore the flexion moment from the lumbrical of only 2% appears to be adequate to prevent clinical clawing of the digit. PMID- 16344188 TI - Comparison of Swanson and Sutter metacarpophalangeal arthroplasties in patients with rheumatoid arthritis: a prospective and randomized trial. AB - PURPOSE: To perform a prospective and randomized comparison of the clinical outcome of patients with rheumatoid arthritis who had Swanson or Sutter implant replacement arthroplasty of the metacarpophalangeal joints. METHODS: There were 45 patients (3 men, 42 women) and 49 hands; a total of 75 Swanson and 99 Sutter implants were inserted. The mean time between surgery and the final follow-up control visit was 58 months (range, 37-80 mo). Preoperative and postoperative measurements were performed including active extension and flexion, correction of ulnar deviation, and strength. RESULTS: There was no statistically significant difference between groups with regard to active extension deficit correction. Mean active flexion decreased less in the Sutter group than in the Swanson group but difference between the groups was statistically significant in only the index finger. At the final follow-up examination no significant differences existed between the groups in the correction of ulnar deviation or arc of motion. Grip strengths, chuck pinch, and thump-to-fingertip grip strengths did not improve in either of the groups. CONCLUSIONS: In this study clinical results showed no significant difference between the groups with the single exception of the amount of index finger metacarpophalangeal joint flexion. PMID- 16344189 TI - Long-term results of silicone wrist arthroplasty in patients with rheumatoid arthritis. AB - PURPOSE: The surgical treatment of the rheumatoid wrist is key in managing the affected hand. Wrist fusion is often the treatment of choice in cases of severe destruction and deformation although most patients would prefer a motion preserving procedure. The implantation of a wrist prosthesis might be an alternative to partial arthrodesis for selected cases. In this series we analyzed the long-term results (minimum follow-up period, 10 y) of the Swanson silicone spacer for the wrist in patients with rheumatoid arthritis. METHODS: Sixteen patients with rheumatoid arthritis with 18 silicone spacers for the wrists were reviewed after a minimum follow-up period of 10 years (average, 15 y). Subjective evaluation, clinical examination, and radiographic analysis were included. An additional 9 patients (9 wrists) were interviewed by telephone. RESULTS: In 12 of the patients the subjective result was good or very good, mostly because of adequate pain relief. The average range of motion for flexion (average, 28 degrees )/extension (average, 15 degrees ) was 43 degrees with a wide variation within the series. Radiologically all wrists had diminished residual carpal height at follow-up evaluation and 9 of the wrists had evidence of osteolysis and foreign-body granuloma. The initial good correction of the ulnar translation of the wrist was lost partially in the follow-up period (1.1 vs 4.0 mm). Three of the patients needed surgical revision within the follow-up period; all were converted to wrist fusion. CONCLUSIONS: These long-term results suggest that the silicone wrist spacer still may be considered as an alternative to wrist fusion or more complex wrist joint prostheses in patients with rheumatoid arthritis, especially in severe cases and in patients with low demands. In the long term osteolysis caused by foreign-body granulation is to be expected and has to be considered. PMID- 16344190 TI - Type 0 ulnar longitudinal deficiency. AB - PURPOSE: To describe the characteristics of type 0 ulnar longitudinal deficiency (ULD) in which deficiencies are present in the hand and carpus without involvement of the forearm or elbow. METHODS: A retrospective chart, radiograph, and clinical photograph review (1960-2005) of patients previously diagnosed with ectrodactyly, hand hypoplasia, or ULD was performed to evaluate for a diagnosis of ULD isolated to the hand. RESULTS: Thirteen extremities were identified. Three extremities had complete absence of the small-finger ray (phalanges and metacarpal) and 6 extremities had complete absence of the ring- and small-finger rays. Four hands showed hypoplasia of the small finger, 3 in conjunction with a ring- and small-metacarpal synostosis and 1 in isolation. Three extremities also had radial-sided hypoplasia or aplasia of the rays. Additional common findings included simple syndactyly, delta phalanx, and carpal fusions, most commonly of the capitohamate joint. CONCLUSIONS: Patients with isolated ulnar-sided hand deficiency such as ectrodactyly of the ring and/or small fingers or synostosis of the small metacarpal of the ring finger in the presence of a normal forearm may be diagnosed as having type 0 ULD. We propose that type 0 be added to the current classification systems for ULD representing those extremities with deficiencies isolated to the hand. PMID- 16344191 TI - Causative microorganisms in surgically treated pediatric hand infections. AB - PURPOSE: The causative organisms in many pediatric musculoskeletal infections are distinct from their adult counterparts. We investigated the organisms found in surgically treated hand infections in the pediatric population. METHODS: We reviewed the hospital charts and computer records of all patients who had surgical intervention for infections of the forearm, wrist, and hand from 1996 through 2002 at The Children's Hospital of Boston. RESULTS: Thirty-eight surgical procedures were performed on 31 children for 32 different upper-extremity infections. One patient was treated for 2 separate infections. In the 32 primary procedures 7 cultures grew mixed aerobic/anaerobic organisms, 19 grew aerobic organisms only, 6 grew no organisms, and 2 patients had no cultures. One culture from a secondary surgery was positive for Candida parapsilosis. Staphylococcus aureus was isolated in 15 of the primary cases and was the primary organism in 12. Five of 7 mixed infections were associated with paronychia; 2 of the paronychial infections were complicated by flexor tenosynovitis. Chronic nail biting caused 2 infections and 1 was associated with thumb sucking. CONCLUSIONS: In the adult population S aureus is the primary organism cultured in 50% to 80% of cases and anaerobic or mixed infection may occur in upward of 29% of patients. In this series S aureus was the primary organism in only 37%. Group A Streptococcus pyogenes was present in 20% of cases. The percentage of children with a mixed aerobic/anaerobic infection was similar to that found in adults; however, the presumed route of introduction was digital sucking/biting. Based on the number of anaerobic/mixed infections without predisposing factors we recommend a broad-spectrum antibiotic regimen including anaerobic coverage as the initial antibiotic in the setting of pediatric hand infections. PMID- 16344192 TI - Reverse osseofasciocutaneous radial forearm flap for thumb reconstruction: a flap design and case series. AB - PURPOSE: To report the outcome of the modified design of the osseofasciocutaneous radial forearm flap. The flap was modified because of problems associated with the use in thumb reconstruction. METHODS: We performed retrospective review of all patients who sustained nonreplantable thumb amputations who had this procedure between August 2000 and March 2005. The patients' ages ranged between 24 and 39 years. Demographic data and details of the level of amputation and alternative surgical procedures were reviewed. Eight male patients were identified and constituted the study cohort. The size of flap, length of the radius harvested, time to union, sensibility (moving 2-point discrimination test), grip and pinch strengths, complications, and patient outcome were determined. RESULTS: All patients who had the butterfly design of the reverse flow osseofasciocutaneous radial forearm flap had an adequate and painless soft tissue padding on the tip of the reconstructed thumb that provided good protection for the distal stump of the vascularized bone graft. Patients did not experience strictures at the bottom of the reconstructed tubes. CONCLUSIONS: The butterfly design is a useful modification of the skin paddle of the reverse osseofasciocutaneous radial forearm flap for thumb reconstruction. The design offers an easy way to reconstruct a tube shape with adequate padding on the tip and no terminal scar. In addition it prevents the constricting effect of the circumferential scar on the button of the reconstructed tube. PMID- 16344193 TI - Innervated reverse island flap based on the end dorsal branch of the digital artery: surgical technique. AB - Fingertip or pulp loss of the fingers is observed frequently in unskilled workers. To reconstruct a sensate fingertip or pulp we designed the innervated reverse island flap based on the end dorsal branch of the digital artery, which was harvested from the dorsum of the middle phalanx. The sensation of the fingertip or pulp was re-established through coaptation of the proper branches of the digital dorsal nerves to the digital nerves. Three fingertip or pulp defects were reconstructed with this technique. All patients achieved satisfactory functional and cosmetic results. The mean follow-up time was 7.7 months. The average size of the flaps was 1.6 x 1.8 cm. The average static 2-point discrimination and moving 2-point discrimination of the flaps were 4.6 mm and 3.0 mm, respectively. The technique we applied seems to be an excellent option for 1 stage reconstruction of fingertip or pulp defects. PMID- 16344194 TI - Radiation exposure in hand surgery: mini versus standard C-arm. AB - PURPOSE: The use of intraoperative fluoroscopy in hand surgery is common. Two types of fluoroscopic units are available: the mini C-arm and the standard C-arm. There is little literature on the radiation exposure from the mini C-arm, therefore, the primary goal of this study was to quantify and compare the amount of radiation exposure to members of the surgical team (surgeon, first assistant, nurse, anesthesiologist) using both standard and mini C-arms in a simulated wrist surgery setup. Mini C-arm positioning was also examined to determine the safest configuration to minimize radiation exposure to surgeons. METHODS: Radiation dosimeters were used to test 2 commercially available fluoroscopy units in a simulated wrist surgery setup with a cadaveric upper extremity. Several different configurations of the C-arms were tested to determine radiation exposure rates to surgeons and the operating room staff. RESULTS: The mean in-beam radiation exposures with the use of the mini and standard C-arms were 3,720 mR/h and 6,540 mR/h, respectively. The mini C-arm had universally less radiation exposure than the standard C-arm in the clinical configurations tested. The safest configuration of mini C-arm use to minimize radiation exposure was with the surgeon standing on the image intensifier side of the unit as compared with the source side. Mini C-arm radiation exposure to the hands, groin, chest, and thyroid of the operating surgeons were well below the National Council of Radiation Protection and Measurement's annual dose limits. CONCLUSIONS: In the clinical configurations tested in this study the mini C-arm had lower radiation exposures than the standard C-arm. To reduce radiation exposure maximally surgeons should stand behind the lead-encased image intensifier and should use techniques to reduce exposure. PMID- 16344195 TI - Radiation exposure in hand surgery. PMID- 16344196 TI - Recurrent enchondroma protuberans: a case report. AB - Enchondroma protuberans is a rare benign tumor. We present a case of late recurrence of a previously reported enchondroma protuberans in the hand. This was treated successfully with curettage and calcium carbonate grafting. PMID- 16344197 TI - Stacking of a dermal regeneration template for reconstruction of a soft-tissue defect after tumor excision from the palm of the hand: a case report. AB - Excision of tumors from the hand often leaves tissue defects with exposed tendons or neurovascular structures that require coverage. Various types of free and pedicled grafts have been described for use in these situations. We present a patient who had a tumor excision in the hand followed by wound coverage with a stacked dermal regeneration template. A 50-year-old man presented with a mass over the palm of the hand. He had an incisional biopsy procedure, the results of which suggested malignancy. He then had wide excision with planned temporary skin coverage with a dermal regeneration template. The dermal template incorporated nicely. With adequate skin coverage the palmar defect still was substantial in terms of depth. This was raised with layering or stacking of the template followed by a split-thickness skin graft. Excellent wound healing and coverage of the defect ultimately were achieved. Additionally the patient went on to obtain full range of motion. Stacking of a dermal regeneration template coupled with split-thickness skin grafting was used to fill a soft-tissue defect over the median and nerve and flexor tendons after wide tumor excision. PMID- 16344199 TI - Dynamic external fixation. PMID- 16344201 TI - Acute lung injury and acute respiratory distress syndrome in the pediatric patient. AB - When the lung becomes injured by direct or indirect assault, the body unleashes a massive immune-modulated initiative. The pediatric patient who experiences acute lung injury is at considerable risk for developing life-threatening respiratory compromise. By understanding the mechanisms of illness in these complex patients, the critical care nurse can connect the science of therapy with the art of providing care that minimizes oxygen expenditure and maximizes oxygen delivery. PMID- 16344202 TI - The experience of a broken heart. AB - Infants who have congenital heart disease (CHD) are born with a "broken heart." Health care professionals are presented with the challenge of caring for the infant's physiologically defective heart as well as the emotional heart of the parents as they begin to accept this diagnosis. This study allowed parents to share this experience through phenomenology as an inquiry approach. These descriptions will assist in providing a more complete and deeper understanding of the parent's experience of CHD. Thus, all will know "the experience of a broken heart." PMID- 16344203 TI - Clinical judgments about endotracheal suctioning: what cues do expert pediatric critical care nurses consider? AB - Making accurate and timely judgments based on multiple ways of knowing is an essential skill in critical care nursing practice. Studies have proposed that positive patient outcomes are linked to expert judgments in a variety of critical care situations; however, little is known about clinical judgments related to specific critical care nursing interventions. This article presents a qualitative nursing research study which examined the cues that expert pediatric critical care nurses used in making clinical judgments about suctioning intubated and ventilated, critically ill children. The participants' words and actions attest that the 'sensing' and 'thinking' of the process of cue use, are interwoven with, and integral to, the 'doing,' which is the process of skilled performance. PMID- 16344204 TI - Diabetic ketoacidosis in the pediatric ICU. AB - Diabetic ketoacidosis (DKA), a pathophysiologic, life-threatening process that results from uncontrolled diabetes mellitus-induced hyperglycemia, is seen frequently in the pediatric ICU. This article reviews the pathophysiology, management, goals of treatment, and nursing implications of the child who is diagnosed with DKA. Facets particular to the pediatric population are reviewed. PMID- 16344205 TI - Death in the pediatric ICU: caring for children and families at the end of life. AB - The need to improve care for children and families at the end of life is acknowledged widely. This article reviews current research concerning end-of-life care in the pediatric ICU. How children die, how decisions are made, management of the dying process, and parent and caregiver experiences are major themes. Gaps in current knowledge are identified, and suggestions are made for future research. PMID- 16344206 TI - Pediatric fluid and electrolyte balance: critical care case studies. AB - The care of the critically ill infant or child often is complicated further by disruptions in fluid or electrolyte balance. Prompt recognition of these disruptions is essential to the care of these patients. This article provides an overview of the principles of fluid and electrolyte balance in the critically ill infant and child. Imbalances in fluid homeostasis and imbalances in sodium, potassium, and calcium homeostasis are presented in a case study format. PMID- 16344207 TI - Low cardiac output syndrome: identification and management. AB - Low cardiac output syndrome (LCOS) is a clinical condition that is caused by a transient decrease in systemic perfusion secondary to myocardial dysfunction. The outcome is an imbalance between oxygen delivery and oxygen consumption at the cellular level which leads to metabolic acidosis. Although LCOS is observed most commonly in patients after cardiac surgery, it may present in various disease processes resulting in cardiac dysfunction. This article provides an overview of the determinants involved in oxygen transport, the physiologic factors influencing cardiovascular function, the assessment of hemodynamic variables, the etiology of LCOS, and management strategies, including a brief review of some pharmacologic agents that are used in the treatment of low cardiac output. PMID- 16344208 TI - Nutritional assessment and enteral support of critically ill children. AB - Critical care nurses play an important role in feeding of critically ill children. Many procedures and caregiving interventions, such as placement of feeding tubes, registration of gastric retention, observation and care of the mouth, and administration of nutrition (enteral or parenteral), are within the nursing domain. This article discusses nutritional assessment techniques and enteral nutrition in critically ill children. PMID- 16344209 TI - Pediatric poisonings: recognition, assessment, and management. AB - Poisoning represents one of the most common medical emergencies encountered in young children in the United States, and accounts for a significant proportion of emergency room visits for the adolescent population. Poisoning is a significant and persistent cause of morbidity and mortality in children and adolescents. The scope of toxic substances involved in poisoning is broad, and requires health care providers to have an extensive knowledge of signs and symptoms of poisoning and specific therapeutic interventions and antidotes. Most children who ingest poisons suffer no harm; however, health care providers must recognize, assess, and manage those exposures that are most likely to cause serious injury, illness, or death and initiate appropriate management to minimize the physical injury that may occur. PMID- 16344210 TI - Management of the pediatric postoperative cardiac surgery patient. AB - Optimal management of the postoperative pediatric cardiac surgical patient requires a thorough understanding of patient anatomy, physiology, surgical repair or palliation, and clinical condition. This necessitates a dedicated team of clinicians including skilled nurses, physicians, and respiratory therapists specialized in the care of patients who have complex congenital heart disease. This article provides an overview of the multisystemic risk factors and consequences associated with cadiopulmonary bypass and cardiac surgery. An evaluation of cardiac hemodynamics and a review of major organ systems are included. Essential assessment information and interventional strategies for managing the pediatric postoperative cardiac surgery patient are detailed. PMID- 16344211 TI - Pediatric sepsis: the infection unto death. AB - A significant percentage of pediatric patients admitted to an ICU have an infectious disease process. Many infants and children go on to develop sepsis, a major cause of death in the intensive care unit. Caring for these children presents a collaborative challenge because of the multifactorial etiology and the complicated pathophysiology. This article focuses on the specific implications of sepsis for infants and children. PMID- 16344212 TI - Safety in the pediatric ICU: the key to quality outcomes. AB - Patient safety is a major concern in the pediatric ICU. The acuity has never been higher, patient needs are extremely complex, and the margin for error is small. The concentration on safety needs to revolve around designing safe systems and processes. This article discusses communication, patient identification, catheter related bloodstream infections, unplanned extubations, restraints and medication administration. The health care system of the future must be transparent, making safety information to insurers, patients and health care providers easily available. PMID- 16344213 TI - Adverse responses: sedation, analgesia and neuromuscular blocking agents in critically ill children. AB - Advanced practice nurses (APNs) prescribe sedation, analgesia, and neuromuscular blocking agents in the management of critically ill children. Although most children are unscathed from the use of the medications, some suffer adverse responses. This article elucidates adverse responses to these medications for the APN, including withdrawal syndrome, muscle weakness, decreased gastric motility, corneal abrasions, and costs associated with these morbidities. PMID- 16344214 TI - Enhancing sibling presence in pediatric ICU. AB - Critical illness of a child affects all members of the family, including well brothers and sisters. Stress in their lives results from changes in parental behaviors, caregiving arrangements, and family relationships. These changes, along with limited understanding and information about the crisis, create feelings of confusion, loneliness, jealousy, and sadness, as well as physical symptoms and behavior changes. Educating pediatric ICU nurses about the needs and reactions of well siblings enables them to optimize support to siblings and educate parents. A sibling policy guarantees that choice and support are offered in a consistent and thorough manner to each critically ill child's family; this helps to ensure that the family unit has the abilities to nurture the ill child and other well children. PMID- 16344215 TI - Current management of status asthmaticus in the pediatric ICU. AB - Status asthmaticus (SA) in the pediatric ICU (PICU) can progress to a life threatening emergency. The goal of management is to improve hypoxemia, improve bronchoconstriction, and decrease airway edema through the administration of continuous nebulized beta2 adrenergic agonist with intermittent anticholinergics, corticosteroids, and oxygen. Adjunctive therapies, such as magnesium, methylxanthines, intravenous beta-agonists, heliox, and noninvasive ventilation should be considered in the child who fails to respond to initial therapies. The restoration of adequate pulmonary functions, resolution of airway obstruction, and avoidance of mechanical ventilation should guide management. This article reviews the pathophysiology, assessment, and management of the child who has SA in the PICU to provide the critical care nurse with current information to facilitate optimal care. PMID- 16344216 TI - Tsunami: response to a disaster. AB - On December 26, 2004, a devastating earthquake occurred in the Indian Ocean very near to Sumatra's coast. The Belgian Association for Pediatrics assembled a medical team of 30 volunteers from 4 hospitals to assist with disaster relief. They traveled to Indonesia, set up a rudimentary care facility, and worked with teams from many countries. In a disaster situation, critically ill children who require mechanical ventilation and inotropic support, perish for lack of equipment or adequate follow-up care. Disaster teams are told to focus on surgery, infected wounds, dehydration, and oral rehydration. This article tells one story of disaster relief efforts and proposes an established team of pediatricians to respond to disaster situations in the future. PMID- 16344217 TI - Short- and long-term skeletal relapse after mandibular advancement surgery. AB - This study analyzes short- and long-term skeletal relapse after mandibular advancement surgery and determines its contributing factors. Thirty-two consecutive patients were treated for skeletal Class II malocclusion during the period between 1986 and 1989. They all had combined orthodontic and surgical treatment with BSSO and rigid fixation excluding other surgery. Of these, 15 patients (47%) were available for a long-term cephalography in 2000. The measurement was performed based on the serial cephalograms taken preoperatively; 1 week, 6 months and 14 months postoperatively; and at the final evaluation after an average of 12 years. Mean mandibular advancement was 4.1 mm at B-point and 4.9 mm at pogonion. Representing surgical mandibular ramus displacement, gonion moved downwards 2 mm immediately after surgery. During the short-term postoperative period, mandibular corpus length decreased only 0.5 mm, indicating that there was no osteotomy slippage. After the first year of observation, skeletal relapse was 1.3 mm at B-point and pogonion. The relapse continued, reaching a total of 2.3 mm after 12 years, corresponding to 50% of the mandibular advancement. Mandibular ramus length continuously decreased 1 mm during the same observation period, indicating progressive condylar resorption. No significant relationship between the amount of initial surgical advancement and skeletal relapse was found. Preoperative high mandibulo-nasal plane (ML-NL) angle appears to be associated with long-term skeletal relapse. PMID- 16344218 TI - A randomized trial assessing the effectiveness of different concentrations of isotretinoin in the management of lichen planus. AB - The aim of our 10-year study was to test the effectiveness of topical therapy based on 0.18% isotretinoin, comparing it with that most frequently used, i.e. at 0.05% concentration. Seventy patients with an established diagnosis of oral lichen planus were involved in the study. The patients were randomly divided into two groups, and the drug was administered topically at 0.05% and 0.18% concentrations. The drug at the higher concentration, according to the same protocol, was administered to the patients who did not benefit from the therapy at the lower concentration. None of the cases of reticular lichen planus showed clinical or histological improvement. In contrast, the atrophic-erosive forms showed a significant improvement, both clinical and histological: in 26 patients (at 0.18% concentration) and in nine patients (at 0.05% concentration), the symptoms, as well as the erosions or ulcers observed, disappeared. The disappearance of dysplasic phenomena was observed at 0.18% concentration. Topical application of the drug was accompanied by an increase in soreness and pain, as well as greater sensitivity to hot foods. However, these side effects were transitory, and considered acceptable by the patients. The proposed therapeutic protocol was effective towards highly active atrophic-erosive oral lichen planus with dysplasic phenomena, which is the form of the disease at higher risk of malignant evolution. PMID- 16344219 TI - Frontal sinus obliteration and craniofacial reconstruction with platelet rich plasma in a patient with fibrous dysplasia. AB - Frontal sinus obliteration is accomplished with many materials such as fat, bone, muscle and alloplasts. Fat obliteration is very common but not devoid of morbidity. An alternative method of treatment with PRP platelet rich plasma and human bank bone or autogenous cortical shavings is presented for obliteration and craniofacial repair. A patient with severe symptomatic fronto-orbital fibrous dysplasia underwent reconstruction with PRP. After an uneventful recovery there was complete remission of the symptoms and early bone formation; 18 months after surgery the patient remains asymptomatic and free from sequels. PRP associated to an allograft or to cortical bone shavings seems to be a safe and simple treatment with the advantage of the absence of donor site surgery and morbidity. PMID- 16344220 TI - Robotic movement elicits automatic imitation. AB - Recent behavioural and neuroimaging studies have found that observation of human movement, but not of robotic movement, gives rise to visuomotor priming. This implies that the 'mirror neuron' or 'action observation-execution matching' system in the premotor and parietal cortices is entirely unresponsive to robotic movement. The present study investigated this hypothesis using an 'automatic imitation' stimulus-response compatibility procedure. Participants were required to perform a prespecified movement (e.g. opening their hand) on presentation of a human or robotic hand in the terminal posture of a compatible movement (opened) or an incompatible movement (closed). Both the human and the robotic stimuli elicited automatic imitation; the prespecified action was initiated faster when it was cued by the compatible movement stimulus than when it was cued by the incompatible movement stimulus. However, even when the human and robotic stimuli were of comparable size, colour and brightness, the human hand had a stronger effect on performance. These results suggest that effector shape is sufficient to allow the action observation-matching system to distinguish human from robotic movement. They also indicate, as one would expect if this system develops through learning, that to varying degrees both human and robotic action can be 'simulated' by the premotor and parietal cortices. PMID- 16344221 TI - Disparities in health care in racial and ethnic minorities. PMID- 16344222 TI - Measurement of left ventricular volumes and function using O-15-labeled carbon monoxide gated PET. PMID- 16344223 TI - Can the ST segment be saved? PMID- 16344224 TI - Assessing progression or regression of CAD: the role of perfusion imaging. PMID- 16344225 TI - Measurement of left ventricular volumes and function with O-15-labeled carbon monoxide gated positron emission tomography: comparison with magnetic resonance imaging. AB - BACKGROUND: Positron emission tomography (PET) with inhaled oxygen 15-labeled carbon monoxide (CO) is used as a marker of myocardial blood pool. Only a limited number of studies with small numbers of patients have reported on the assessment of left ventricular (LV) volumes by use of O-15-labeled CO. The aim of this study was to compare LV volumes and function as measured by routinely acquired blood pool images by use of gated O-15-labeled CO PET with the reference technique, cardiovascular magnetic resonance imaging (MRI). METHODS AND RESULTS: Thirty-four subjects with a varying degree of LV function were studied. LV end-diastolic volume (LVEDV), LV end-systolic volume (LVESV), and LV ejection fraction (LVEF) were determined by both MRI and gated PET by use of O-15-labeled CO. Volumes were comparable with respect to LVEDV (196 +/- 83 and 192 +/- 91 mL, respectively; P = not significant). LVESV, however, was slightly overestimated by PET (119 +/- 85 and 136 +/- 94 mL, respectively; P < .05), resulting in a significant underestimation of LVEF (44% +/- 19% and 35% +/- 18%, respectively; P < .05). Observed correlations for LVEDV, LVESV, and LVEF were 0.90, 0.96, and 0.86, respectively (all P < .01). CONCLUSIONS: Gated O-15-labeled CO PET measurements of LVEDV, LVESV, and LVEF show good correlation with MRI over a wide range of LV volumes during routinely acquired blood pool images. LVEF, however, may be underestimated compared with MRI. PMID- 16344226 TI - Comparison of Tc-99m sestamibi SPECT with fractional flow reserve in patients with intermediate coronary artery stenoses. AB - BACKGROUND: Myocardial single photon emission computed tomography (SPECT) is an established noninvasive method for the assessment of the functional significance of coronary artery stenoses. Intracoronary pressure measurements to determine fractional flow reserve (FFR) are increasingly performed during coronary angiography whenever an immediate decision regarding possible intervention is required. We hypothesized that the regional summed difference score (SDSr), reflecting reversible perfusion defects in the myocardial supply area of the FFR target vessel, would be the best predictor of an abnormal FFR in patients without prior myocardial infarction. Otherwise, a regional summed stress score (SSSr) should be the best predictor of an abnormal FFR in patients with prior myocardial infarction for different patient subgroups with coronary artery disease. METHODS AND RESULTS: In this study 50 patients (mean age, 65 +/- 9.1 years; 18 women) with coronary artery disease and a 50% to 75% coronary stenosis (target vessel) were prospectively investigated. Dobutamine myocardial SPECT was performed as a single-day stress/rest protocol by use of technetium 99m sestamibi. For image interpretation, semiquantitative analysis was conducted by calculating SSSr and SDSr. Within 8 (+/-14.9) days, coronary angiography was performed and FFR was calculated by use of a pressure wire (normal FFR, > or = 0.75). The mean FFR of all patients was 0.78 +/- 0.14. Of 50 patients, 17 had an FFR lower than 0.75 in the target vessel. Receiver operating characteristic analysis identified an SDSr of 1 or greater and an SSSr of 3 or greater as the best threshold values for predicting ischemic FFR. Sensitivity, specificity, and negative and positive predictive values of SDSr and SSSr for the detection of FFR values lower than 0.75 in the target vessel were 80%, 76%, 53%, and 92%, respectively, and 70%, 93%, 78%, and 90%, respectively, in patients without prior myocardial infarction and 57%, 50%, 67%, and 40%, respectively, and 100%, 50%, 78%, and 100%, respectively, in patients with prior myocardial infarction. Weak correlation was found between the single values of FFR with both SDSr and SSSr for the different patient subgroups. CONCLUSION: Among the dobutamine myocardial scintigraphy variables studied, SDSr was the best predictor of an abnormal FFR (cutoff value of 0.75) in patients without prior myocardial infarction. As assumed, SSSr was the best predictor of an abnormal FFR in patients with prior myocardial infarction in the target region. PMID- 16344227 TI - Prognostic value of myocardial perfusion SPECT versus exercise electrocardiography in patients with ST-segment depression on resting electrocardiography. AB - BACKGROUND: The value of exercise-induced ST-segment depression for the prognostic evaluation of patients with 1 mm of ST depression or greater on the resting electrocardiogram is controversial. METHODS AND RESULTS: Patients who underwent exercise myocardial perfusion single photon emission computed tomography (MPS) and had resting ST depression of 1 mm or greater with a nondiagnostic exercise electrocardiographic response (n = 1122) were followed up for 3.4 +/- 2.3 years. Those with paced rhythm, pre-excitation, left bundle branch block, or myocardial revascularization within the first 60 days after MPS were excluded. Additional exercise-induced ST-segment depression was considered significant if > or = 2 mm MPS was scored semiquantitatively by use of a 20 segment model of the left ventricle; the percentage of myocardium involved with stress defects (% myo) was derived by normalizing to the maximal possible score of 80. Hard events were defined as nonfatal myocardial infarction or cardiac death. A Cox analysis was used to determine independent predictors of hard events among clinical, exercise, and nuclear variables. Hard event rates increased as a function of % myo for either patients with exercise-induced ST depression (1.4%/y for normal MPS vs 4.1%/y for % myo >10%, P < .03) or those without it (0.7%/y for normal MPS vs 3.0%/y for % myo >10%, P = .0001). Age, diabetes mellitus, shortness of breath as the presenting symptom, and % myo were independent predictors of hard events. Exercise-induced ST depression was predictive of hard events only when it was 3 mm or greater. The presence and extent of perfusion defects, reflected in the % myo, had incremental prognostic value over clinical variables and also over all degrees of exercise-induced ST depression. CONCLUSIONS: Although MPS effectively risk-stratifies patients with resting ST depression of 1 mm or greater, the prognostic value of exercise-induced ST depression is limited in these patients, with a small added risk when severe (> or = 3 mm). PMID- 16344228 TI - Incremental prognostic value of combined perfusion and function assessment during myocardial gated SPECT in patients aged 75 years or older. AB - BACKGROUND: Perfusion and functional data obtained during gated single photon emission computed tomography (SPECT) have proven prognostic value in the middle aged patient population. The aim of this study was to investigate whether perfusion and functional cardiac gated SPECT data have prognostic value in patients aged 75 years or older. METHODS AND RESULTS: We studied clinical and gated SPECT predictors of cardiac and all-cause death in 294 patients aged 75 years or older with known or suspected coronary artery disease who were referred for tetrofosmin cardiac gated SPECT imaging. Summed perfusion scores were calculated in a 17-segment model by use of commercially available software (4D MSPECT). Left ventricular functional data were calculated by use of QGS gated SPECT software. The median age of the study population was 78 years (range, 75-91 years). There were 160 men (54%) and 134 women (46%). During a median follow-up of 25.9 months (range, 1.8-36 months), 47 patients (16%) died (27 cardiac deaths). In a multivariate Cox proportional hazards regression analysis, the summed rest score (chi2 gain = 8.0, P = .009), transient ischemic dilatation index (chi2 gain = 6.3, P = .012), and resting left ventricular ejection fraction (chi2 gain = 7.0, P = .030) were independent predictors of all-cause death. The summed rest score (chi2 gain = 8.2, P = .004) and resting end-systolic volume (chi2 gain = 13.7, P = .005) were independent predictors of cardiac death. CONCLUSIONS: This study showed that gated SPECT left ventricular functional data assessed during myocardial gated SPECT provide independent and incremental information above clinical and perfusion SPECT data for the prediction of cardiac and all-cause death in patients aged 75 years or older referred for myocardial SPECT imaging. PMID- 16344229 TI - Prognostic value of stress myocardial perfusion imaging in octogenarian population. AB - BACKGROUND: The prognostic value of myocardial perfusion imaging (MPI) in the very elderly population has not been addressed specifically. The aim of this study was to examine the characteristics of the octogenarian population referred for MPI and the prognostic value of the procedure in terms of patient gender. METHODS: The study sample consisted of 162 consecutive patients (61 women, 101 men) of mean age 83 +/- 3 years (range, 80-90) who underwent stress MPI. The duration of follow-up was 45+/-12 months. Outcome measures were cardiac-related death, and nonfatal myocardial infarction (MI). RESULTS: Thirty-one major cardiac events (19%) were recorded, including 26 cardiac deaths and 5 MI events, in 6 of the 61 women (10%) and 25 of the 101 men (25%) (p < 0.03). The univariate predictors of cardiac death or MI, except of known coronary artery disease (CAD), were MPI variables: left ventricular (LV) dilatation, increased lung uptake, abnormal scan, and the presence and extent of myocardial ischemia. However, the only predictors for major cardiac events were: LV dilatation (OR = 6.9, 95% CI 2.7-17.4, p < 0.0001) and ischemia by scan (OR = 2.75, 95% CI 1.09-6.96, p < 0.03). The Kaplan Meier curve demonstrated significant differences in survival between patients with or without LV dilatation and patients with or without ischemia. CONCLUSIONS: LV dilatation and myocardial ischemia were useful predictors of cardiac death and MI in octogenarian patients with CAD or with suspicion of CAD. PMID- 16344230 TI - Clinical validation of SPECT attenuation correction using x-ray computed tomography-derived attenuation maps: multicenter clinical trial with angiographic correlation. AB - BACKGROUND: Nonuniform attenuation artifacts cause suboptimal specificity of stress single photon emission computed tomography (SPECT) myocardial perfusion images. In phantoms, normal subjects, and patients suspected of having coronary artery disease (CAD), we evaluated a new hybrid attenuation correction (AC) system that combines x-ray computed tomography (CT) with conventional stress SPECT imaging. METHODS AND RESULTS: The effect of CT-based AC was evaluated in phantoms by assessing homogeneity of normal cardiac inserts. AC improved homogeneity of normal cardiac phantoms from 11% +/- 2% to 5% +/- 1% (P < .001). Attenuation-corrected normal patient files were created from 37 normal subjects with a low likelihood (<3%) of CAD. The diagnostic performance of AC for detection of CAD was evaluated in 118 patients who had stress technetium 99m sestamibi or tetrofosmin stress SPECT imaging and coronary angiography. SPECT images with and without AC were interpreted by 4 blinded readers with different interpretative attitudes. Overall, AC improved the diagnostic performance of all readers, particularly the normalcy rate. The degree of improvement depended on interpretative attitude. Readers prone to high sensitivity or with less experience had the greatest gain in the normalcy rate, whereas a reader prone to higher specificity had improvements in sensitivity and specificity but not the normalcy rate. Importantly, improvement of one diagnostic variable was not associated with worsening of other variables. CONCLUSION: CT-based AC of SPECT images consistently improved overall diagnostic performance of readers with different interpretive attitudes and experience. CT-based AC is well suited for routine use in clinical practice. PMID- 16344231 TI - Onset of left ventricular mechanical contraction as determined by phase analysis of ECG-gated myocardial perfusion SPECT imaging: development of a diagnostic tool for assessment of cardiac mechanical dyssynchrony. AB - BACKGROUND: A count-based method using technetium-99m sestamibi electrocardiography-gated myocardial perfusion single photon emission computed tomography imaging has been developed to extract the left ventricular (LV) regional phase of contraction (onset of mechanical contraction [OMC]) throughout the cardiac cycle. This study was performed to develop OMC normal databases and dynamic OMC displays for assessment of cardiac mechanic dyssynchrony. METHODS AND RESULTS: LV regional phases were extracted from 90 enrolled normal subjects (45 men and 45 women) by use of the Emory Cardiac Toolbox and then submitted to statistical analysis to generate the normal databases. The LV OMC wave was dynamically propagated over the perfusion polar map by blackening either sequential phase bins or all past phases. The developed OMC normal databases consisted of peak phase (134.5 degrees +/- 14.3 degrees for men and 140.2 degrees +/- 14.9 degrees for women), phase SD (14.2 degrees +/- 5.1 degrees for men and 11.8 degrees +/- 5.2 degrees for women), and phase histogram bandwidth (38.7 degrees +/- 11.8 degrees for men and 30.6 degrees +/- 9.6 degrees for women), skewness (4.19 +/- 0.68 for men and 4.60 +/- 0.72 for women), and kurtosis (19.72 +/- 7.68 for men and 23.21 +/- 8.16 for women). Both statistical analysis and dynamic OMC displays were incorporated into a user interface as a diagnostic tool. CONCLUSION: The OMC normal databases and dynamic OMC displays should help clinicians evaluate cardiac mechanic dyssynchrony. Prospective clinical trials are needed to validate whether this tool can be used to select patients with severe heart failure symptoms who might benefit from cardiac resynchronization therapy. PMID- 16344232 TI - Sinus arrest during adenosine stress testing in liver transplant recipients with graft failure: three case reports and a review of the literature. AB - BACKGROUND: Nuclear stress imaging is used frequently to evaluate patients with end-stage liver disease who are being considered for orthotopic liver transplantation. METHODS AND RESULTS: We present three patients who, following graft failure, developed sinus arrest during adenosine stress testing performed in anticipation of repeat liver transplantation. All had undergone uneventful adenosine stress imaging prior to initial transplantation. The mechanisms of action, pharmacokinetics, and pharmacodynamics of adenosine are reviewed, and possible reasons for this phenomenon are discussed. Finally, cautions regarding the use of adenosine and treatment of adenosine-induced sinoatrial and atrioventricular block are reviewed. CONCLUSION: Adenosine should be used with caution in patients following orthotopic liver transplantation due to an increased risk of sinus arrest. Should sinus arrest or atrioventricular block occur, it appears to respond readily to cessation of adenosine infusion and intravenous aminophylline with no significant sequelae. PMID- 16344233 TI - Current and future status on cardiac computed tomography imaging for diagnosis and risk stratification. AB - Computed tomography (CT) permits cross-sectional imaging with high spatial resolution and has, during the past years, undergone tremendous development mainly concerning the temporal resolution. By use of multidetector spiral technology, as well as electrocardiography-gated image acquisition and reconstruction techniques, 16- and 64-slice CT permits visualization of cardiac morphology and function. In this context, however, CT imaging does not play a major clinical role because other imaging methods (mainly echocardiography) usually provide all necessary information. Under certain conditions, multidetector CT also permits visualization of the coronary arteries. Detection of coronary calcification, as well as coronary CT angiography, can provide clinically useful information if applied to suitable patient groups. It is foreseeable that CT angiography will become part of the routine workup in some subsets of patients with suspected coronary artery disease, either alone or in combination with other imaging techniques. Among the limitations of cardiac CT are the requirement of a regular (and preferably low) heart rate, the associated x-ray exposure, and the need for an iodinated contrast agent for most applications. It is important to note that reliable and accurate results will require use of the most advanced CT scanner technology, optimal image quality, and sufficient experience in the acquisition and interpretation of cardiac CT data sets. PMID- 16344234 TI - Cardiac receptor physiology and imaging: an update. PMID- 16344235 TI - Imaging inflammatory cardiomyopathies. PMID- 16344236 TI - Unusual radiotracer uptake in the lower mediastinum on sestamibi perfusion images. PMID- 16344237 TI - Metabolic evidence of myocardial stunning in takotsubo cardiomyopathy: a positron emission tomography study. PMID- 16344238 TI - Cost-effectiveness of myocardial perfusion imaging: a summary of the currently available literature. PMID- 16344239 TI - A novel spectrofluorimetric method for the determination of DNA. AB - A new simple, selective and sensitive fluorescence quenching method was developed to determine nucleic acids (DNA) with the 9-anthracenecarboxylic acid (ACA)-cetyl trimethyl-ammonium bromide (CTAB) system. The fluorescence intensity of ACA was decreased by the addition (CTAB). However, the fluorescence intensity of the system increased dramatically when DNA was added to the solution. The fluorescence enhancement is probably based on the DNA interaction with CTAB. Under the optimum conditions, the changes of fluorescence intensity in the absence and presence of nucleic acids was proportional to the concentration of nucleic acids over the range 0.08-1.0 microg mL(-1) for CT (calf thymus) DNA or FS (fish sperm) DNA. Its detection limits are 0.02 microg mL(-1) for CT DNA and 0.019 microg mL(-1) for FS DNA. Based on this approach, a new quantitative method for DNA assay is presented in this paper. PMID- 16344240 TI - ESR and electrochemical study of 5-nitroindazole derivatives with antiprotozoal activity. AB - The electrochemistry of 3-alkoxy- and 3-hydroxy-1-[omega-(dialkylamino)alkyl]-5 nitroindazole derivatives were characterized using cyclic voltammetry in DMSO. The nitro reduction process was studied and this was affected by the acid moieties present in these compounds. A nitro anion self-protonation process was observed. This phenomenon was studied by cyclic voltammetry in presence of increasing amount of NaOH. The reactivity of the nitro anion radical of these derivatives with glutathione was also studied by cyclic voltammetry. The oxidizing effect of glutathione is supported by the parallel decrease of the anodic peak current and increase of the cathodic peak in the cyclic voltammograms, corresponding to the wave of the nitro anion radical from uncharged species with the addition of glutathione. Nitro anion radicals obtained by electrolytic reduction of these derivatives were measured and analyzed in DMSO using electron spin resonance spectroscopy. PMID- 16344241 TI - Copper(II)-selective fluorimetric bulk optode membrane based on a 1-hydroxy-9,10 anthraquinone derivative having two propenyl arms as a neutral fluorogenic ionophore. AB - A new optical chemical sensor has been developed for the selective determination of copper(II) ions in aqueous solutions. The reversible sensing system was prepared by incorporating 1-hydroxy-2-(prop-2'-enyl)-4-(prop-2'-enyloxy)-9,10 anthraquinone (AQ) as a neutral Cu2+-selective fluoroionophore in the plasticized PVC membrane with potassium tetrakis(p-chlorophenyl borate) as an anionic additive. The response of the sensor is based on the fluorescence quenching of AQ by Cu2+ ions. At a pH 5.5, the proposed sensor displays a calibration response for Cu2+ over a wide concentration range of 1.0 x 10(-2) to 1.0 x 10(-6) M, with a relatively fast response of less than 40 s. In addition to high stability and reproducibility, the sensor shows a unique selectivity towards Cu2+ ion with respect to common co-existing cations. The proposed fluorescence optode was applied successfully to the determination of copper(II) in black tea samples. PMID- 16344242 TI - Effects of Yb concentration on the spectroscopic properties of Yb: Y3Al5O12. AB - Yb:YAG single crystals with Yb doping concentration 5.4, 16.3, 27.1, 53.6, and 100 at.% were grown by the Czochralski process. The effects of Yb concentration on the absorption spectra (190-1100 nm), fluorescence spectra under 940 nm and X ray excitation were studied. The concentration quenching of fluorescence was observed when the Yb doping concentration reaches to as high as 27.1at.% for Yb:YAG. Under 940 nm excitation, the influence of the self-absorption at 969 and 1029 nm on the fluorescence spectra is not evident when the Yb doping concentration is as high as 27.1at.%. However, it can greatly change the shape of fluorescence spectra of Yb:YAG when the Yb doping concentration reaches to above 53.6 at.%. PMID- 16344243 TI - ESR and theoretical studies of trimer radical cations of coronene. AB - Highly resolved ESR spectra of monomer, dimer and trimer radical cations of coronene (C24H12) were observed at room temperature for a solution of 1,1,1,3,3,3 hexafluoro-2-propan-2-ol (HFP) containing thallium(III) trifluoroacetate as oxidant. The spectra consisting of multiple lines with isotropic 1H-hyperfine splitting (hfs) constants of 0.0766 mT (24H) and 0.013 mT (6H) were attributable to a mixture of the dimer with the trimer radical cations, (C24H12)2+ and (C24H12)3+. For (C24H12)2+, the 1H-hfs constant agreed well with the reported value, 0.077 mT. However, for (C24H12)3+, the values were significantly different from the reported ones, 0.117 mT (12H) and 0.020 mT (24H), by Ohya Nishiguchi et al. [H. Ohya-Nishiguchi, H. Ide, N. Hirota, Chem. Phys. Lett. 66 (1979) 581], but rather similar to those reported by Willigen et al. [H. van Willigen, E. De Boer, J.T. Cooper, W.F. Forbes, J. Chem . Phys. 49 (1968) 1190]. In conflict with Willigen's report, however, no ESR line broadening which has been ascribed to a low stationary concentration of (C24H12)3+ was detected. Based on ab initio MO calculations for benzene as a compact model of C24H12, the structure of (C24H12)3+ was investigated in terms of the observed 1H-hfs constants. A staggered sandwich C(2v) structure was suggested being at the "global" minimum for the benzene trimer cation. In the structure, the unpaired electron spin is predominantly localized to the central ring, which is qualitatively in agreement with the previous ESR results of (C24H12)3+ by Ohya-Nishiguchi et al. In addition, as a "local" minimum, the benzene trimer was indicated to have a slipped sandwich Cs structure, which is less stable by ca. 19 kJ mol(-1) than the "global" minimum. In this structure, the unpaired electron spin was nearly equally distributed on both the central and one of the two side C24H12 molecules. The observed 1H-hfs constants were possibly attributable to the (C24H12)3+ cation with the analogous slipped sandwich Cs structure. PMID- 16344244 TI - Phase matchable semiorganic NLO material for frequency doubling: L-Arginine tetrafluoroborate. AB - L-Arginine tetrafluoroborate (C6H15N4(+)O2BF4(-)) is a semiorganic crystalline material exhibiting second harmonic generation (SHG) down to the UV region. The material was synthesized and characterized by elemental analysis, FT-NMR and FT IR spectra. The lower cutoff wavelength was found to be 270 nm, with a good optical transmittance window covering UV-vis-NIR region. The thermal stability of the compound was determined by TG-DTA curves. The powder SHG efficiency was measured using a Q-Switched Nd:YAG laser assembly by a modified Kurtz and Perry powder technique and its phase matching property was established. PMID- 16344245 TI - Cobalt(II) complexes of biologically active glutathione: spectroscopic and molecular modeling studies. AB - Cobalt(II) complexes of reduced glutathione (GSH) of general composition Na[Co(L)(X)].nH2O (where H2L = GSH; X = Cl-, NO3-, NCS-, CH3CO2-, HCO2-, ClO4- and n = 0-4) have been synthesized and characterised by elemental analyses, vibrational spectra, electronic spectra, magnetic susceptibility measurements, thermal studies and molecular modeling studies. Electronic spectra indicate planar geometry for all the complexes. Infrared spectra indicate the presence of H2O molecules (except perchlorate complex) in the complexes that has been supported by TG/DTA. The room temperature magnetic moment values for all complexes lie in the range of 2.60-2.80 BM range indicating departure from spin only values due to second order Zeeman effect. Thermal decomposition of all the complexes proceeds via first order kinetics. The Na[Co(L)(Cl)].2H2O complex has the minimum activation energy and Na[Co(L)(CH3CO2)].3H2O has the maximum activation energy. The molecular modeling calculation for energy minimization optimizes geometry of the metal complexes. PMID- 16344246 TI - Classification of the hydrogen-bonding species in a series of novel hydrazide based azobenzene derivatives investigated by two-dimensional correlation infrared spectroscopy and molecular modeling. AB - Classification of hydrogen-bonding species in a series of novel hydrazide modified p-methoxyazobenzene derivatives, 4-{n-[4-(4-methoxy-phenylazo)-phenoxy] alkoxy}-benzoic acid hydrazide (Dn, n = 3, 6, 10) are performed in the present study. Temperature-dependent infrared (IR) spectra of Dn have been measured to investigate the thermal stability of the weak intermolecular interactions, such as hydrogen bonding among hydrazide moieties, pi-pi stacking among aromatic groups, and hydrophobic interaction between alkyl chains. In order to reveal the hydrogen bonding formed between NH, NH2, and CONH groups efficiently, two dimensional (2D) correlation spectra have been constructed in the thermal sensitive spectral regions of (a) 3500-3100 cm(-1) and (b) 1700-1450 cm(-1), separately, and it have also been constructed between these two spectral regions. Based on the experimental data, the ab initio computational models have been developed to the proposed patterns of hydrogen bonding related to intermolecular interactions in Dn. The intermolecular hydrogen bondings and molecular alignments patterns result from both the experimental data and the computational models are performed for D3, D6, and D10, respectively, in the present study. PMID- 16344247 TI - Studies of the defect structures of the trigonal Cr3+-Vc centers in fluoroperovskites from EPR and optical spectra. AB - The EPR parameters (zero-field splitting D and g factors g(parallel), g(perpendicular)) and the first excited state splitting Delta(2E) of the trigonal Cr3+-Vc centers (Vc denotes the cation vacancy in a C3 axis) in Cr3+-doped fluoroperovskites KMgF3, KZnF3, CsCdF3, CsCaF3, RbCdF3 and BaLiF3 are studied from the high-order perturbation formulas. From the studies, the defect structures (characterized by the vacancy-induced displacements Delta x1 of Cr3+ ion and Delta x2 of the three F- ion between Cr3+ and Vc) of these trigonal Cr3+ Vc centers are determined. It is found that the signs of displacements Delta x1 and Delta x2 are consistent with the expectations based on the electrostatic interactions, and the magnitudes of Delta x1 and Delta x2 for Cr3+ in the inverse perovskite BaLiF3 are larger than those for Cr3+ in the classical perovskites. The results are discussed. PMID- 16344248 TI - Calculation of electric dipole intensity parameter to explore some interaction between hard metal ions Pr(III) and Nd(III) with pi-electron density of butene 1,4 and butyne-1,4-diols in non-aqueous solutions: an absorption spectral study. AB - Pr(III) and Nd(III) are hard acceptors in HSAB (hard and soft acid base) sense and hence are known to exhibit practically a little affinity towards electrons. At the same time these metal ions show strong preference for oxygen donor chelating ligands. The ligands chosen for this study are structurally related diols, viz. butane-1,4, butene-1,4 and butyne-1,4-diols which form identical seven membered chelate ring by coordinating to metals in a bidentate manner through oxygen on 1 and 4 positions of the diol molecules. Complexation of these diols with Pr(III) and Nd(III) was carried out in DMF, CH3OH, CH3CN and their equimolar binary mixtures using comparative absorption spectrophotometry of 4f-4f transitions. The variation of oscillator strengths (P) of different 4f-4f bands as well as the magnitude and variation of Judd-Ofelt electric dipole intensity parameters (T lambda, lambda = 2, 4, 6) was discussed. They correlate the interaction between the metal 4f-orbitals of Pr(III) and Nd(III) with the pi electron densities of the double and triple bonds present in butene-1,4 and butyne-1,4-diols, respectively. The value of empirical intensity parameter [T lambda(complex)/T lambda (aquo)] was calculated and its plot against oscillator strength (P) is drawn. PMID- 16344249 TI - Near IR overtone spectral investigations of liquid phase imidazole and the blue shifting hydrogen bonds. AB - The near infrared vibrational overtone absorption spectrum of imidazole dissolved in carbon tetrachloride is examined in the region Delta v = 2, 3 and 4. Instead of the standard hydrogen bonding, unusual blue-shifted hydrogen bonds are observed in NH positions. This observation is in agreement with the recent conclusions drawn from theoretical studies of the coupling character between the Imidazole-Imidazolium complex, which is strongly hydrogen bonded. The NH local mode mechanical frequency values and anharmonicity values obtained from fitting the overtones are analysed. The anharmonicity of blue-shifted hydrogen bonded NH stretching bands is greater compared to those of the free NH stretching bands. It is contrary to the decrease in anharmonicity due to the standard red shifting of NH bonds. PMID- 16344250 TI - Structural and spectral perspectives of a novel thiosemicarbazone synthesized from di-2-pyridyl ketone and 4-phenyl-3-thiosemicarbazide. AB - A new thiosemicarbazone, HL is synthesized from di-2-pyridyl ketone and 4-phenyl 3-thiosemicarbazide and structurally and spectrochemically characterized. 1H NMR, 13C NMR, COSY, HMQC and IR spectra of the compound are studied and the proton magnetic resonance spectrum reveals some unprecedented observations. The thione form is predominant in the solid state, as supported by the crystal structure and IR data, while a thiol-thione equilibrium is proposed in the solution state by NMR studies. The compound crystallizes into a monoclinic lattice with space group C2/c and the ZE conformation is exhibited by the thiosemicarbazone. Intra- and intermolecular hydrogen-bonding interactions give rise to a two-dimensional packing in the crystal lattice. PMID- 16344251 TI - Sensitized spectrophotometric determination of Cr(III) ion for speciation of chromium ion in surfactant media using alpha-benzoin oxime. AB - A simple and accurate micellanized spectrophotometric method for determination of trace amounts of Cr(III) ion in tab and top water and a synthetic mixture has been described. The micellar method is based on effect of organized molecular assemblies such as micelles in spectrophotometric measurement due to their effect on the systems of interest. The ability of micellar system in solubilizing of sparingly soluble ligand or complexes has been used for increasing figures of merit of an analytical method. Due to solubility increasing in aqueous media requirement for a primary extraction can be eliminated. Using the alpha-benzoin oxime (alpha-BO) spectrophotometric determination of Cr(III) ion has been performed and results are compared. The spectrophotometric determination of Cr(III) ion using alpha-BO in the presence of non-ionic surfactant Triton X-100 has been performed. The influence of type and amount of surfactant, pH, complexation time and amount of ligand were examined. Finally, the repeatability, accuracy and the effect of interfering ions on the determination of Cr(III) ion was evaluated. The proposed methods successfully with recovery yield of almost 100% have been applied to the rapid and simple determination of Cr(III) ion in the real samples. There is a good agreement between methods and atomic absorption spectrometry. The Beers law is obeyed over the concentration range of 0.1-13.7 microg mL(-1) for micellar media. The detection limit is 0.8 ng mL(-1). The molar absorptivity of complex is 5350 L mol(-1) cm(-1). PMID- 16344252 TI - Detailed 1H and 13C NMR structural assignment of three biologically active lignan lactones. AB - In this paper we present a complete 1H and 13C NMR spectral analysis of three lignan lactones (methylpluviatolide, dimethylmatairesinol and hinokinin) by the use of techniques such as COSY, HMQC, HMBC and J-resolved. Complete assignment and all homonuclear hydrogen coupling constant measurements were performed, providing enough data also to the confirmation of the relative stereochemistry. PMID- 16344253 TI - Variability of prostate brachytherapy pre-implant dosimetry: a multi institutional analysis. AB - PURPOSE: To conduct a multi-institutional comparison of prostate brachytherapy pre-implant dosimetry of Pd-103 and I-125. METHODS AND MATERIALS: Eight experienced brachytherapists submitted Pd-103 and I-125 monotherapeutic and boost pre-implant dosimetry plans for central review. All 32 plans were calculated using the same transrectal ultrasound volumetric study. Seeds of any strength were acceptable, but were restricted to Theraseed Model 200 (Theragenics Inc., Buford, GA) and Oncura Oncoseed Model 6711 (Oncura, Plymouth Meeting, PA). The dosimetric analysis included evaluation of target volume, target to prostate ratio, target length, number of needles, seed activity, number of seeds, total activity, total activity divided by treatment planning volume, the use of extracapsular seeds, and average treatment margins (defined as the perpendicular distance between the prostate capsule and the 100% isodose line). Prostate coverage was defined in terms of V(100)/V(150)/V(200)/V(300) and D(100)/D(90)/D(50), whereas urethral dosimetry consisted of UV(100)/UV(150)/UV(200) and UD(90)/UD(50). RESULTS: The mean planning target volume to prostate volume ratio varied dramatically (mean 1.29, range 0.99-1.76) with the target length ranging from 3.5 to 4.5 cm. Although the prostate V(100) was >95% in all cases, the V(150) ranged from 29.9% to 92.1% and the V(200) from 6.72% to 52.5%. The urethral V(100) was 100% in all cases with six of the eight brachytherapists limiting the UV(150) to <3%. However, the median urethral dose varied by up to 50%. Treatment margins also varied significantly (average 3.98 mm, range 0.32-7.68 mm). All brachytherapists used extracapsular seeds with five implanting >25% of the seeds in extracapsular locations (range 6.4-58.2%). In addition, significant variability existed in the number of needles, number of seeds, and seed strength. CONCLUSIONS: This study highlights the substantial variability that exists regarding target volume, seed strength, dose homogeneity, treatment margins, and extracapsular seed placement, although prostate brachytherapy prescription doses are uniform. The standardization of pre-implant dosimetry is essential for meaningful multi-institutional comparisons of biochemical outcomes and morbidity. PMID- 16344254 TI - Dosimetric quantifiers for low-dose-rate prostate brachytherapy: is V(100) superior to D(90)? AB - PURPOSE: The purpose of the present report is to describe the relationship between two dosimetric quantifiers (V(100) and D(90)) and freedom from biochemical recurrence (FFBR) in a cohort of men treated with low-dose-rate prostate brachytherapy (LDRPB) alone. METHODS AND MATERIALS: One hundred three men were treated with LDRPB alone between September 1997 and December 1999. All men had histologically confirmed clinically localized prostate cancer. Fifty-nine percent of the cohort had low-risk disease (defined as PSA<10, Gleason <7, and T stage /=10, Gleason>/=7, or T stage T2b). The prescription dose was 144Gy according to the Task Group 43 formalism. LDRPB was performed jointly by a radiation oncologist and a urologist. Dosimetric quantifiers (D(90), V(100)) were calculated from a CT scan performed 1 month after LDRPB. Biochemical recurrence was defined according to the ASTRO Consensus Definition. FFBR was estimated using the product-limit method. Disease-specific and treatment variables were examined as putative covariates for FFBR using the proportional hazards regression method. Univariate and multivariate methods were used. All p values are two sided. RESULTS: The median followup for the entire cohort is 61 months. The median followup of patients at risk for biochemical failure is 66 months. The median D(90) is 129Gy (range 47-221Gy), and the median V(100) is 86% (range 51-99%). Thirteen men have developed evidence of biochemical relapse at a median of 25 months (range 6-42 months). The 5-year estimate of FFBR for the entire cohort is 87% (95% CI 80 94%). On univariate analysis, disease-specific variables found to be significantly associated with FFBR included pretreatment PSA and percent positive biopsies. When considered as a continuous variable, each of the dosimetric quantifiers was associated with FFBR (V(100): p=0.007; D(90): p=0.05). D'Amico risk group classification is highly predictive of FFBR after LDRPB (HR 5.68, p=0.003). Multivariate analysis indicated that each dosimetric quantifier was independently associated with FFBR, but due to the high degree of correlation (Pearson correlation coefficient 0.94, p<0.0001) between the dosimetric quantifiers both could not be included simultaneously in the model. In the two models explored, V(100) was at least as good as D(90) in predicting FFBR. CONCLUSIONS: Dosimetric quantifiers (V(100) and D(90)) are independent predictors of FFBR after treatment with LDRPB alone. In our experience, V(100) seems to be at least equivalent (and perhaps superior) to D(90) for predicting FFBR. PMID- 16344255 TI - Chest wall dose in MammoSite breast brachytherapy: radiobiologic estimations of late complication risk based on dose-volume considerations. AB - PURPOSE: To estimate the risk of late effects in women treated with MammoSite brachytherapy (MBT), the balloon catheters of which were placed near the ribs. METHODS AND MATERIALS: Upon reviewing 93 plans, 16 patients (17%) treated with MBT were considered to have received a high chest wall dose (>or=120% isodose line in contact with a rib). A dose-volume histogram was generated for this rib, and its distance from the MBT balloon measured. Using the linear quadratic equation, the equivalent dose, delivered in 10 fractions, to the dose that causes a 5% and 50% risk of rib late effects at 5 years using 2Gy per fraction, was calculated to be 37 and 44Gy, respectively. The rib volume receiving greater than or equal to these doses (V37 and V44) was correlated to the balloon-to-rib distance. Chest wall signs, symptoms, and radiologic findings for all 16 patients were recorded. RESULTS: The median balloon-to-rib distance was 4.8mm. The median values of V37 and V44 were 13.5% and 3.3%, respectively. All patients with a V37>or=15% and V44>or=5% had a minimum balloon-to-rib distance of <5mm. Two patients reported treatment-related chest wall tenderness (both had balloons placed <5mm from the chest wall), but neither presented with radiologic complications. CONCLUSIONS: Sixteen patients considered to receive relatively high chest wall doses had less than one-third of their primary rib volume being exposed to the estimated TD 5/5 and TD 50/5 doses. Therefore, we estimate the risk of late effects in women treated with MBT, the balloon catheters of which placed near the ribs were negligible, and believe that MBT remains a safe and effective treatment for selected patients with early stage breast cancer. PMID- 16344256 TI - Perioperative high-dose-rate brachytherapy in soft tissue sarcomas of the extremity and superficial trunk in adults: initial results of a pilot study. AB - PURPOSE: This study was undertaken to determine the feasibility of perioperative high-dose-rate brachytherapy (PHDRB) as an alternative to standard low-dose-rate brachytherapy in adult patients with soft tissue sarcomas. METHODS AND MATERIALS: Twenty-five adult patients with soft tissue sarcomas of the extremities or the superficial trunk were treated with surgical resection and PHDRB (16, 24, or 32Gy) for negative, close/microscopically positive, or grossly involved surgical resection margins, respectively. External beam radiation therapy (45Gy) was added postoperatively. Adjuvant chemotherapy with ifosfamide and doxorubicin was given to patients with high-grade tumors. RESULTS: Resection margins were negative in 6 patients (24.0%), close/microscopically positive in 18 (72.0%), and grossly involved in 1 (4.0%). One patient (4.0%) with wound dehiscence needed reoperation. Three patients failed in the anatomical region treated, but relapse sites were not adjacent to the area treated with PHDRB. After a median followup of 23.2 months (range 2.8-48.0), the 4-year local and regional control rates were 100% and 80.5%, respectively. Four-year overall survival was 78.2%. CONCLUSIONS: Locoregional control and survival results are encouraging for this high-risk patient population. PHDRB results seem to be similar to those obtained with low dose-rate brachytherapy. PMID- 16344257 TI - Longitudinal evaluation of patients with cancer in the oral tongue, tonsils, or base of tongue--does interstitial radiation dose affect quality of life? AB - PURPOSE: To evaluate health-related quality of life (HRQL) in patients with oral tongue, tonsil, or base of tongue cancer in a prospective longitudinal study and explore correlations between HRQL scores and interstitial radiation dose, dose rate, and volume of implant. METHODS AND MATERIALS: Ninety patients with oral tongue cancer (n=30) and tonsil or base of tongue cancer (n=60) were assessed with the European Organization of Research and Treatment of Cancer Quality of Life Questionnaire Core-30 and the European Organization of Research and Treatment of Cancer Head and Neck module at diagnosis, and after 3, 12, and 36 months of starting treatment. RESULTS: The HRQL of all patients decreased during treatment. Most HRQL scores returned to baseline values after 3 years; however, 60% of patients with oral tongue cancer and 80% with tonsil and base of tongue cancer reported problems with dry mouth and half of the patients with tonsil and base of tongue cancer reported problems with swallowing solid food at the 3-year followup. No correlations between brachytherapy quality indices and HRQL scores were found. CONCLUSIONS: Patients with oral tongue, tonsil, or base of tongue cancer reported significant problems with dry mouth and swallowing solid food throughout this 3-year followup study. PMID- 16344258 TI - Intraoperative (125)I Vicryl mesh brachytherapy after sublobar resection for high risk stage I non-small cell lung cancer. AB - PURPOSE: To assess the feasibility and outcomes of (125)I Vicryl mesh brachytherapy after sublobar resection in stage I non small cell lung cancer (NSCLC) patients with poor pulmonary function. METHODS AND MATERIALS: Between January 1997 and July 2004, patients with poor cardiopulmonary reserve who had stage IA and IB (T1-2 N0 M0) NSCLC and a forced expiratory volume in 1s (FEV(1)) of > micro=0.6L were considered for limited surgical resection either by an open or video-assisted thoracoscopic procedure and for a subsequent (125)I Vicryl mesh brachytherapy implant. Mediastinal and hilar lymph node staging was performed routinely in all patients. After clear margins were obtained grossly and on frozen section, a single-plane (125)I implant was designed to encompass a plane consisting of the staple line and a 2-cm margin of surrounding visceral pleura. The implant was introduced through the surgical incision and sutured to the visceral pleura. A prescribed dose of 100-120 y was delivered to a volume within 0.5 cm rom the plane of the implant. Follow-up orthogonal films or CTs were obtained for dosimetric analysis. Kaplan-Meier analyses were used to estimate the local control, locoregional control, and overall survival rates. RESULTS: Of the 110 patients, 65 had stage IA and 45 had stage IB NSCLC. The mean preoperative FEV(1) was 47% of the predicted volume. With a median follow-up of 11 months (range 1-68 months), there were four recurrences within the radiation volume. The estimated 5-year local (in-field) control, locoregional control, and overall survival rates were 90%, 61%, and 18%, respectively. CONCLUSION: Vicryl mesh brachytherapy after sublobar resection for high-risk stage I NSCLC patients is a feasible procedure, which results in an excellent local (in-field) control rate. PMID- 16344259 TI - Image-based intracavitary brachytherapy in the treatment of inoperable uterine cancer: individual dose specification at specific anatomical sites. AB - PURPOSE: With advances in imaging studies, dose specification for uterine cancer can be defined at specific anatomical sites such as the myometrium or the serosal surface rather than at arbitrary points or milligram-hours. This report presents our experience with image-based brachytherapy for inoperable uterine cancer. METHODS AND MATERIALS: Eight patients with organ-confined uterine cancer (2 Stage I GI, 3 Stage I G2, 3 Stage I G3) underwent definitive radiation therapy because of poor medical condition. All the patients underwent a CT or MRI scan of the pelvis before intracavitary application. Based on the size of the uterine cavity, a single-channel intrauterine applicator was selected for a small uterus, and a multiple-channel intrauterine applicator was used for a large uterus. A CT (n=5) or MRI (n=3) scan of the pelvis was performed with the applicator in place in addition to orthogonal pelvic films. Individualized dose specification was 75Gy to the midmyometrium and limited to 50Gy to the serosal surface of the uterus based on imaging information. RESULTS: Four patients with Stage I G1-2 disease had intracavitary brachytherapy alone. Four patients with Stage I G2-3 disease were treated with a combination of external pelvic radiation and intracavitary brachytherapy. Six patients had low-dose-rate brachytherapy, and 2 patients had high-dose-rate brachytherapy. Five patients had single-channel intrauterine brachytherapy, and 3 patients had multiple-channel brachytherapy. Based on the measurements of the uterine wall thickness by the imaging studies, the dose specification was prescribed to 1.5 cm lateral to the central axis of the uterus in 4 patients, 2.0 cm in 3 patients, and 2.5 cm in 1 patient. The medium followup time after radiation treatment was 38 months. Six patients are alive without evidence of disease, and 2 patients died of other causes. All patients had local control without major side effects. CONCLUSIONS: Image-based brachytherapy based on individualized dose specification at specific anatomical sites can be done easily and provides excellent local control for inoperable uterine cancer. PMID- 16344260 TI - Improving patient-specific dosimetry for intravascular brachytherapy. AB - PURPOSE: Accurate patient-specific dosimetry in intravascular brachytherapy (IVBT) is generally difficult due to the extremely high-dose gradient, complexity of treatment device, and patient-specific geometry (e.g., calcification, stent, curvature, movement of target). The purpose of this study is to analyze quantitatively and systematically the dose effects of calcification, stent, guidewire, and source curvature on clinical dosimetry in an IVBT procedure, and propose a method that can be used to assess these effects in routine clinical practice. METHODS AND MATERIALS: Monte Carlo techniques were used to calculate 3 D dose distribution in both homogeneous and inhomogeneous media for three most commonly used IVBT sources: (90)Sr beta (Novoste), (192)Ir gamma (Cordis/Best), and (32)P beta (Guidant). Dosimetric perturbations in the presence of metallic stents, calcified plaques, metallic guide wires, and source curvature were studied for situations commonly encountered in the clinic. The importance of each of these perturbations and their practical influence on patient-specific dosimetry were analyzed. Factors (plaque, stent, guidewire, and curvature) that may be used to correct/reduce these perturbations were introduced to prevent dosimetric cold spots during IVBT. Practical methods of using these correction factors are proposed. RESULTS: Dose perturbations are significant due to the presence of source curvature, metallic stents, calcified plaques, and metallic guide wires, especially for beta sources. These perturbations can be as high as 30% under normal clinical conditions, although they can be much higher in extreme situations. Empirical relationships of plaque factor with the thickness of calcified plaque, stent factor with stent metallic surface area, guidewire with guidewire thickness, and curvature factor with the bending angle are derived. These relationships are found to be useful in improving clinical dose accuracy in IVBT treatment planning or dose evaluation after treatment. CONCLUSIONS: Significant dose perturbations due to the presence of source curvature, metallic stents, calcified plaques, and guide wires have been found in IVBT for in-stent restenosis. Because it has been reported that, with the current prescriptions for IVBT, higher doses consistently improve treatment outcomes, the empirical method derived from this work can be used to assess cold spots dosimetrically, thus improving patient-specific dosimetry for IVBT. PMID- 16344261 TI - Quantitative evaluation of acute mucosal reaction to interstitial brachytherapy using color histograms. AB - PURPOSE: Color histograms of tongue photographs were used to make a quantitative evaluation of acute mucosal reaction to high-dose-rate fractionated interstitial brachytherapy. METHODS AND MATERIALS: Six patients with oral cavity cancer were analyzed with two methods (color histogram and vinyl plate). Using a photograph of the tongue, we calculated the histogram of each pixel of 1mm(2) of the tongue surface. The results were compared with the physician's inspection and the radiation dose. RESULTS: The histogram showed a difference between the reactive area (erythema and pseudomembranous mucositis [PM]) and the nonreactive area. The relationship between inspection and the histogram showed a good correlation, although the histogram tended to change earlier than the subjective judgment. The peak time was 12-13 days after delivery of the final fraction of brachytherapy. The minimum dose required to cause PM was 3.3-3.6Gy per fraction (total: 29.7 32.4Gy). CONCLUSIONS: Our new method of using color histograms for quantitative evaluation is helpful in the analysis of the relationship between mucosal reaction and radiation dose. The minimum dose causing PM was considered to be about 30Gy per nine fractions. PMID- 16344262 TI - Gravitropism in leaves of Arabidopsis thaliana (L.) Heynh. AB - In higher plants, stems and roots show negative and positive gravitropism, respectively. However, current knowledge on the graviresponse of leaves is lacking. In this study, we analyzed the positioning and movement of rosette leaves of Arabidopsis thaliana under light and dark conditions. We found that the radial positioning of rosette leaves was not affected by the direction of gravity under continuous white light. In contrast, when plants were shifted to darkness, the leaves moved upwards, suggesting negative gravitropism. Analysis of the phosphoglucomutase and shoot gravitropism 2-1 mutants revealed that the sedimenting amyloplasts in the leaf petiole are important for gravity perception, as is the case in stems and roots. In addition, our detailed physiological analyses revealed a unique feature of leaf movement after the shift to darkness, i.e. movement could be divided into negative gravitropism and nastic movement. The orientation of rosette leaves is ascribed to a combination of these movements. PMID- 16344263 TI - Reaction mechanism and substrate specificity for nucleotide sugar of mammalian alpha1,6-fucosyltransferase--a large-scale preparation and characterization of recombinant human FUT8. AB - FUT8, mammalian alpha1,6-fucosyltransferase, catalyzes the transfer of a fucose residue from the donor substrate, guanosine 5'-diphosphate (GDP)-beta-L-fucose, to the reducing terminal GlcNAc of the core structure of asparagine-linked oligosaccharide via an alpha1,6-linkage. FUT8 is a typical type II membrane protein, which is localized in the Golgi apparatus. We have previously shown that two neighboring arginine residues that are conserved among alpha1,2-, alpha1,6-, and protein O-fucosyltransferases play an important role in donor substrate binding. However, details of the catalytic and reaction mechanisms and the ternary structure of FUT8 are not understood except for the substrate specificity of the acceptor. To develop a better understanding of FUT8, we established a large-scale production system for recombinant human FUT8, in which the enzyme is produced in soluble form by baculovirus-infected insect cells. Kinetic analyses and inhibition studies using derivatives of GDP-beta-L-fucose revealed that FUT8 catalyzes the reaction which depends on a rapid equilibrium random mechanism and strongly recognizes the base portion and diphosphoryl group of GDP-beta-L-fucose. These results may also be applicable to other fucosyltransferases and glycosyltransferases. PMID- 16344264 TI - Chemical and histochemical analysis of 'Quatre Saisons Blanc Mousseux', a Moss Rose of the Rosa x damascena group. AB - BACKGROUND AND AIMS: Moss roses are old garden roses covered with a mossy growth on flower pedicel and calyx. This moss releases a pine-scented oleoresin that is very sticky and odoriferous. Rosa x centifolia 'muscosa' was the first moss rose to be obtained by bud-mutation but, interestingly, R. x damascena 'Quatre Saisons Blanc Mousseux' was the first repeat-blooming cultivar, thus interesting breeders. In the present study, the anatomy of these sports (i.e. bud-mutations) is characterized and the volatile organic compounds (VOCs) produced by the moss versus the petals are identified. They are compared between the two lines and their respective parents. METHODS: Anatomy of the moss is studied by environmental scanning electron microscopy and histochemical light microscopy. Sudan Red IV and Fluorol Yellow 088 are used to detect lipids, and 1-naphthol reaction with N,N-dimethyl-p-phenylenediamine to detect terpenes (Nadi reaction). Head-space or solid/liquid extraction followed by gas chromatography and mass spectrometry are used to identify VOCs in moss, trichomes and petals. KEY RESULTS: Moss of the two cultivars has the same structure with trichomes on other trichomes but not exactly the same VOCs. These VOCs are specific to the moss, with lots of terpenes. An identical VOC composition is found in leaves but not in petals. They are nearly the same in the moss mutants and in the respective wild types. CONCLUSIONS: Sepals of moss roses and their parents have a specific VOC pattern, different from that of the petals. The moss corresponds to a heterochronic mutation with trichomes developing on other trichomes. Such a mutation has probably appeared twice and independently in the two lines. PMID- 16344265 TI - Pollination ecology of Silene acutifolia (Caryophyllaceae): floral traits variation and pollinator attraction. AB - BACKGROUND AND AIMS: The floral display influences the composition of pollinators interacting with a plant species. Geographic and temporal variation in pollinator composition complicates the understanding of the evolutionary consequences of floral display variation. This paper analyses the relationships between Silene acutifolia, a hermaphroditic perennial herb, and its pollinators, based on field studies in the north-west of Spain. METHODS: Studies were conducted over three years (1997-1999). Firstly, the main pollinators of this species were determined for two years in one population. Secondly, pollen limitation in fruit and seed production was analysed by supplementary hand pollinations, and counting the pollen grains and tubes growing in styles for two different-sized populations. Finally, the effect of flower size and number on the rate of visitation and total seed number was examined for 15 marked plants. RESULTS AND CONCLUSIONS: The primary pollinators were long-tongued insects, including Hymenoptera, Lepidoptera and Diptera, but the composition and visitation frequencies differed between years. Pollen limitation occurred in one of the years of study. There was between population variation in the number of pollen grains and pollen tubes found in styles, suggesting pollen limitation in one population. Overall, pollinators visited plants with more open flowers more frequently, and pollinated more flowers within these plants. Conversely, petal and calyx sizes had no effect on insect visitation. Plants with higher rates of visits produced higher number of seeds, suggesting that pollinator-mediated limitation of seed and fruit production may be important in some years. PMID- 16344266 TI - Incorporation of tissue reaction kinetics in a computational fluid dynamics model for nasal extraction of inhaled hydrogen sulfide in rats. AB - Rodents exposed to hydrogen sulfide (H2S) develop olfactory neuronal loss. This lesion has been used by the risk assessment community to develop occupational and environmental exposure standards. A correlation between lesion locations and areas of high H2S flux to airway walls has been previously demonstrated, but a quantitative dose assessment is needed to extrapolate dose at lesion sites to humans. In this study, nasal extraction (NE) of 10, 80, and 200 ppm H2S was measured in the isolated upper respiratory tract of anesthetized rats under constant unidirectional inspiratory flow rates of 75, 150, and 300 ml/min. NE was dependent on inspired H2S concentration and air flow rate: increased NE was observed when H2S exposure concentrations or inspiratory air flow rates were low. An anatomically accurate, three-dimensional computational fluid dynamics (CFD) model of rat nasal passages was used to predict NE of inhaled H2S. To account for the observed dependence of NE on H2S exposure concentration, the boundary condition used at airway walls incorporated first-order and saturable kinetics in nasal tissue to govern mass flux at the air:tissue interface. Since the kinetic parameters cannot be obtained using the CFD model, they were estimated independently by fitting a well-mixed, two-compartment pharmacokinetic (PK) model to the NE data. Predicted extraction values using this PK-motivated CFD approach were in good agreement with the experimental measurements. The CFD model provides estimates of localized H2S flux to airway walls and can be used to calibrate lesion sites by dose. PMID- 16344267 TI - Investigating human cancer etiology by DNA lesion footprinting and mutagenicity analysis. AB - Many genotoxic carcinogens are known to leave unique signatures on cancer-related genes. The signature of carcinogens is manifested by the induction of characteristic mutations at distinctive nucleotide positions along oncogenes and/or tumor suppressor genes. Often, the nucleotide positions, wherein mutations occur, co-localize with the sites of initial DNA damage induced by the respective carcinogens. Thus, DNA damage-targeted mutation can be a predictor of carcinogenicity of genotoxins. Today, genomic sequencing technologies for investigating human cancer etiology are based on DNA-lesion footprinting in conjunction with mutagenicity analysis of genotoxic carcinogens. In this review article, we discuss the ligation-mediated PCR and terminal transferase-dependent PCR, two versatile DNA-lesion footprinting techniques. We highlight the in vitro shuttle vector-based mutation systems for investigating site-specific mutagenicity of carcinogens and the in vivo transgenic rodent mutation systems for exploring DNA damaging and mutagenic properties of carcinogens. We present examples of application of each of these methodologies to human cancer etiology, and provide prospective views on investigations using these technologies for carcinogenicity testing. PMID- 16344268 TI - Joint effect of asthma/atopy and an IL-6 gene polymorphism on lung cancer risk among lifetime non-smoking Chinese women. AB - Recent evidence suggests that inflammatory pathways are important mediators of carcinogenesis. Asthma, allergic rhinitis and atopic dermatitis are clinical manifestations of a systemic atopic disorder, which is associated with airway hyper-responsiveness and inflammation. We examined the effect of a history of asthma/atopy among 132 lung cancer cases (of which 72% were adenocarcinomas) and 163 controls, all of whom were non-smoking Chinese women, in combination with a single nucleotide polymorphism (-634C/G) in the interleukin-6 (IL-6) gene which regulates secretion of a pro-inflammatory cytokine found to be predominant in lung tumour tissue. We observed a slight increase in risk of lung cancer [odds ratio, OR = 1.5, 95% confidence interval (95% CI) = 0.8-2.6] and of adenocarcinoma (OR = 1.6, 95% CI = 0.9-3.1) with asthma/atopy alone. There was no effect of the IL-6 CG/GG genotype on lung cancer risk on its own. Among individuals with both asthma/atopy and the IL-6 -634 G allele, however, risk was increased at least 3-fold (OR = 3.1, 95% CI = 1.2-8.3 for all cancers and OR = 4.2, 95% CI = 1.5-11.6 for adenocarcinomas) relative to individuals with no asthma/atopy and the CC genotype. On stratified analysis, a significant increase in risk with asthma/atopy was restricted to those with the at-risk genotype (Pint < 0.05). Our findings are consistent with the role of chronic inflammation as an aetiologic factor among non-smoking Asian women, and suggest that asthma/atopy is a risk marker for susceptibility to the development of lung cancer. PMID- 16344269 TI - Efficacy of new retinoids in the prevention of mammary cancers and correlations with short-term biomarkers. AB - A number of retinoid X receptor (RXR) agonists have proven to be highly effective in preventing methylnitrosourea (MNU) induced mammary cancers. However, these agonists have side effects; particularly causing an increase in serum triglyceride levels. A series of ligands for RXR were designed based on computer modeling to the ligand binding domain (LBD) of the RXR receptors and on structure activity relationships. The chemopreventive effects of these retinoids were evaluated in the relatively long-term MNU model. As a short-term assay to predict their efficacy, the ability of the retinoids to modulate cell proliferation and apoptosis was also determined in mammary cancers after only 7 days of treatment. The five UAB retinoids evaluated included two Class I UAB retinoids (UAB20, UAB112) and three Class II UAB retinoids (UAB30, 4-methyl-UAB30 and the benzosuberone-analog of UAB30). The previously evaluated RXR agonist targretin and the pan-agonist 9-cis-retinoic acid (9-cis-RA), which interacts with both RAR and RXR receptors, were included as positive agonists known to prevent cancer in the MNU model. In the prevention studies, in which the agents were administered beginning 5 days after MNU until the end of the study, targretin (150 mg/kg diet) and 4-methyl-UAB30 (200 mg/kg diet) were highly effective in decreasing cancer numbers by 75-85%. UAB30 (200 mg/kg diet) and 9-cis-RA (60 mg/kg diet) gave intermediate inhibitions of 60 and 45%, respectively. Targretin (15 mg/kg diet), UAB20 (200 mg/kg diet) and the benzosuberone analog of UAB30 (200 mg/kg diet) showed limited activity by decreasing cancer multiplicity 25-30%, while UAB112 had no effect on mammary cancer multiplicity. A direct correlation was observed between the long-term chemopreventive efficacy of these agents and their ability to decrease cell proliferation in mammary cancers after short-term treatment. Furthermore, the highly effective agents (4-methyl-UAB30 and targretin at 150 mg/kg diet) increased apoptosis 3-5 times, while agents with moderate or limited preventive efficacy failed to significantly increase apoptosis. Although the more effective retinoid treatments increased serum triglycerides 2.5- to 4.0-fold, one moderately effective agent (UAB30) had no significant effect on lipid levels. In summary, a short-term in vivo method has been identified for screening newly synthesized retinoids both for chemopreventive efficacy and for their adverse effect on serum triglycerides. PMID- 16344270 TI - Violacein synergistically increases 5-fluorouracil cytotoxicity, induces apoptosis and inhibits Akt-mediated signal transduction in human colorectal cancer cells. AB - Despite recent additions to the armory of chemotherapeutic agents for colorectal cancer (CRC) treatment, the results of chemotherapy remain unsatisfactory. 5 Fluorouracil (5-FU) still represents the cornerstone of treatment and resistance to its actions is a major obstacle to successful chemotherapy. Therefore, new active agents in CRC and agents that increase the chemosensitivity of cancer cells to 5-FU are still urgently required. Violacein, a pigment isolated from Chromobacterium violaceum in the Amazon river, has a diverse spectrum of biological activities, and represents a novel cytotoxic drug with known antileukemic properties. To assess the suitability of violacein as a chemotherapeutic agent in CRC its cytotoxic effects were evaluated both as a single agent and in combination with 5-FU. Its underlying mechanisms of action were further investigated by studying its effects on the cell cycle, apoptosis and cell survival pathways [phosphatidylinositol-3-kinase/Akt, p44/42 mitogen activated protein kinase and nuclear factor kappaB (NF-kappaB)] in colon cancer cell lines. Violacein inhibits the growth of all four colon cancer cell lines tested. It induces apoptosis, and potentiates the cytotoxic effect of 5-FU in a poorly differentiated microsatellite unstable cell line (HCT116). Violacein causes cell cycle block at G(1), upregulates p53, p27 and p21 levels and decreases the expression of cyclin D1. Violacein leads to dephosphorylation of retinoblastoma protein and activation of caspases and a pancaspase inhibitor abrogates its biological activity. Our data provide evidence that violacein acts through the inhibition of Akt phosphorylation with subsequent activation of the apoptotic pathway and downregulation of NF-kappaB signaling. This leads to the increase in chemosensitivity to 5-FU in HCT116 colon cancer cells. Taken together, our findings suggest that violacein will be active in the treatment of colorectal tumors and offers new prospects for overcoming 5-FU resistance. PMID- 16344271 TI - A proteomic investigation into a human gastric cancer cell line BGC823 treated with diallyl trisulfide. AB - Garlic is generally used as a therapeutic reagent against various diseases worldwide. Although a great effort is made to understand the pharmaceutical mechanisms of garlic and its derivatives, there are many mysteries to be uncovered. Using proteomic means, herein we have systematically studied the responses of protein expression in BGC823 cells, a gastric cancer cell line, induced by diallyl trisulfide (DATS), a major component of garlic derivatives. A total of 41 unique proteins in BGC823 were detected with significant changes in their expression levels corresponding with DATS administration. Of these proteins, five typical ones, glutathione S-transferase-pi (GST-pi), voltage dependent anion channel-1 (VDAC-1), Annexin I, Galectin and S100A11, were further examined by Western blotting, resulting in coincident data with the proteomic evidence. Moreover quantitative real-time RT-PCR experiments offered dynamic data of mRNA expression, indicating the responses of Annexin I and GST-pi expression within a short period after DATS treatment. Interestingly, approximately 50% of DATS-sensitive proteins (19/41) in BGC823 are tightly associated with apoptotic pathways. These proteomic results presented, therefore, provide additional support to the hypothesis that garlic is a strong inducer of apoptosis in tumor cells. PMID- 16344272 TI - Aberrant expression of PDGF ligands and receptors in the tumor prone ovary of follitropin receptor knockout (FORKO) mouse. AB - Although PDGF family members play a vital role in cell proliferation, motility and chemotaxis via activation of structurally similar alpha- and beta-receptors, little is known of their function in ovarian regulation and induction of tumorigenesis. Microarray analyses of ovaries from young follitropin receptor knockout (FORKO) mice that are prone to late ovarian tumors upon aging have revealed significant imbalances in PDGF ligands and receptors. We hypothesized that FSH/FSH-R signaling may exert effects partly by regulation of PDGF the family. To further understand their implications for ovarian tumorigenesis, we studied FORKO ovaries and hormonal regulation of the PDGF family members in normal mice, by using RT-PCR, Q-PCR, immunohistochemistry and western blotting. While PDGF-C and PDGFR-alpha increased, PDGFR-beta mRNA and protein decreased significantly in absence of FSH-R signaling. In the normal ovary, PDGFR-alpha was not affected by gonadotropin (eCG) stimulation but PDGF-C and PDGFR-beta decreased. Administration of estradiol decreased PDGF and their receptors. To further probe the differential regulation of PDGF family members by eCG and estradiol, we co-administered eCG with estrogen antagonist, ICI 182780. Increase in PDGFR-alpha in the absence of estradiol suggests direct effects of FSH signaling. During the estrous cycle in mice PDGF-C, PDGF-D and PDGFR-alpha mRNA levels were higher at the proestrous. By IHC, we report for the first time the localization of PDGF-C, PDGFR-alpha and PDGFR-beta protein in mouse ovarian compartments including the surface epithelium that is also altered in mutants. Immunostaining of PDGFRs increased as the follicle developed to preantral stage and declined thereafter. Thus, FSH modulates PDGF family members, partly via E2, suggesting that loss of FSH-R signaling causes an imbalance of PDGF family members predisposing the abnormal ovarian follicular environment for inducing tumorigenesis in aging FORKO mice. PMID- 16344273 TI - Decreased expression of the human stem cell marker, Rex-1 (zfp-42), in renal cell carcinoma. AB - The Rex-1 (Zfp-42) gene encodes a zinc finger family transcription factor which is highly expressed in mouse and human embryonic stem cells. It is one of several gene markers used to identify human stem cells. While several organs are known to harbor adult human stem cells, the presence and distribution of stem cells in both the normal and neoplastic adult kidney remains largely unknown. In this study we evaluated Rex-1 mRNA and protein expression in normal and malignant kidney tissue specimens from human patients. Rex-1 mRNA expression was determined using both reverse transcription and real-time PCR. REX1 protein expression was assessed by western analysis and immunohistochemistry, using an affinity purified, polyclonal antibody to the REX1 protein. We found that 14 of 15 (93%) non-tumor renal parenchymal specimens demonstrated Rex-1 mRNA, compared with 5 of 14 (36%) renal tumors (P < 0.005). REX1 protein expression was detected in 21 of 23 (91%) non-tumor and in 7 of 19 (37%) tumor specimens (P < 0.001). Furthermore, in six of these seven renal tumor specimens where REX1 protein expression was detected, the levels were at least 3-fold lower than those in adjacent, normal kidney tissue. There were no differences in Rex-1 mRNA or protein expression among the various histologic subtypes of renal tumors (clear cell carcinoma, papillary carcinoma, chromophobe carcinoma and oncocytoma). Immunohistochemical staining confirmed the absence of REX1 in three renal tumor specimens (two clear cell and one papillary carcinoma), while the REX1 protein was detected in a small percentage of proximal tubular cells in normal renal tissue. Immunohistochemical staining of another stem cell marker, OCT4, demonstrated a similar pattern of protein expression in a small percentage of normal renal proximal tubular cells. In summary, we were able to detect Rex-1 mRNA and protein expression in over 90% of normal renal parenchymal specimens and we observed a significant reduction in REX1 expression in renal tumor specimens of all histologic subtypes. PMID- 16344274 TI - ADH3 genotype, alcohol intake and breast cancer risk. AB - Moderate alcohol consumption of approximately 1-2 drinks per day has been associated with a 30-50% increase in breast cancer risk. Individuals differ in their ability to metabolize alcohol through genetic differences in alcohol dehydrogenase (ADH), the enzyme that catalyzes the oxidation of approximately 80% of ethanol to acetaldehyde, a known carcinogen. Individuals differ in their ADH genotype, and one locus in particular (ADH3) is polymorphic in Caucasian populations. Using data from the Long Island Breast Cancer Study Project, we examined whether fast metabolizers of alcohol, as measured by the ADH3(1-1) genotype, have a higher risk of breast cancer from alcohol intake compared with those individuals who are slow metabolizers, but consume similar amounts of alcohol. We combined genotyping information with questionnaire data on 1047 breast cancer cases and 1101 controls and used unconditional logistic regression methods to estimate multivariate-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) between alcohol intake and breast cancer risk. Among individuals homozygous for the fast metabolizing allele (ADH(3)1-1), a lifetime alcohol consumption of 15-30 g/day (approximately 1-2 drinks per day) increased breast cancer risk by 2-fold (OR=2.0, 95% CI=1.1-3.5). In contrast, the increase in risk from a lifetime alcohol consumption of 15-30 g/day was less pronounced in the intermediate and slow metabolizing groups, respectively: ADH3(1-2) (OR=1.5, 95% CI 0.9-2.4) and ADH(3)2-2 (OR=1.3, 95% CI 0.5-3.5). Fast metabolizers who drank 15-30 g/day of alcohol had 2.3 times (95% CI 1.3-4.0) greater risk of breast cancer than non-drinkers who were intermediate or slow metabolizers. This association for fast metabolizers who drank 15-30 g/day was particularly pronounced among premenopausal women (premenopausal women OR=2.9, 95 % CI=1.2 7.1; postmenopausal women OR=1.8, 95% CI=0.9-3.8). These population-based data support the hypothesis that fast metabolizers of alcohol have a higher risk of breast cancer risk, from alcohol intake than slow metabolizers. PMID- 16344275 TI - Folate intake and squamous-cell carcinoma of the oesophagus in Italian and Swiss men. AB - BACKGROUND: Dietary folate has been inversely related to the risk of several cancers. However, studies on the role of dietary folate in oesophageal cancer are scanty. PATIENTS AND METHODS: Using data from a multicentric case-control study conducted in Italy and Switzerland between 1992 and 1999, we investigated the association between dietary folate intake and oesophageal squamous-cell carcinoma (OSCC) among 351 men with incident, histologically confirmed OSCC and 875 hospital controls admitted for acute, non-neoplastic conditions, unrelated to alcohol and smoking consumption. Intake of folate and other nutrients was computed from a validated food frequency questionnaire. RESULTS: The multivariate odds ratios (ORs) of OSCC were 0.68 (95% confidence intervals, CI: 0.46-1.00) for the highest versus the lowest tertile of folate intake, and 0.84 (95% CI: 0.72 0.99) for an increment of folate intake equal to a standard deviation (98 microg/day). The inverse relation was somewhat stronger in strata of high methionine, vitamin B6 and alcohol intake, and did not vary substantially according to age and smoking habits. CONCLUSION: Dietary folate was inversely related to OSCC risk in this population with high alcohol consumption and infrequent use of supplements and multivitamins. PMID- 16344276 TI - Impact of providing booklets about chemotherapy to newly presenting patients with cancer: a randomized controlled trial. AB - BACKGROUND: There has been limited research examining the efficacy of providing written information to cancer patients in southern and eastern European countries. This study investigated the impact of a booklet about chemotherapy on patient satisfaction, quality of life (QoL) and emotional distress, and assessed booklet use. PATIENTS AND METHODS: A total of 145 Greek cancer outpatients prior to commencing chemotherapy completed a questionnaire and were randomized to receive (n = 72) or not receive (n = 73) the booklet. All patients completed the second questionnaire before the following cycle of chemotherapy. RESULTS: Baseline characteristics were well balanced between the two groups. Experimental group patients reported being significantly more satisfied with the information received and care overall than those in the control group, felt significantly more and better informed, and perceived the information received as being clearer and detailed. The intervention produced no benefits in terms of anxiety, depression or QoL. The booklet was read by almost all patients and to a great extent by significant others. The majority considered the booklet useful to read and helpful in recalling chemotherapy-related information. CONCLUSIONS: The current results provide further evidence on the efficacy of information-giving interventions in societies where disclosure of information to cancer patients still remains controversial. PMID- 16344277 TI - Systematic review and meta-analysis of randomised, controlled trials of the timing of chest radiotherapy in patients with limited-stage, small-cell lung cancer. AB - BACKGROUND: We undertook a systematic review and literature-based meta-analysis to determine whether the timing of chest radiotherapy may influence the survival of patients with limited-stage small-cell lung cancer (LS-SCLC). MATERIALS: Eligible randomised controlled clinical trials were identified according to the Cochrane Collaboration Guidelines, comparing different timing of chest radiotherapy in patients with LS-SCLC. Early chest irradiation was defined as beginning within 30 days after the start of chemotherapy. RESULTS: Considering all seven eligible trials, the overall survival at 2 or 5 years was not significantly different between early or late chest radiotherapy. When only trials were considered that used platinum chemotherapy concurrent with chest radiotherapy, a significantly higher 5-year survival was observed when chest radiotherapy was started within 30 days after the start of chemotherapy (2-year survival: OR: 0.73, 95% CI 0.51-1.03, P = 0.07; 5-year survival: OR: 0.64, 95% CI 0.44-0.92, P = 0.02). This was even more pronounced when the overall treatment time of chest radiotherapy was less than 30 days. CONCLUSIONS: There are indications that the 5-year survival rates of patients with LS-SCLC are in favour of early chest radiotherapy, with a significant difference if the overall treatment time of chest radiation is less than 30 days. PMID- 16344278 TI - A randomised cross-over trial comparing patient preference for oral capecitabine and 5-fluorouracil/leucovorin regimens in patients with advanced colorectal cancer. AB - BACKGROUND: Traditionally, metastatic colorectal cancer (MCRC) has been treated with intravenous (i.v.) 5-fluorouracil/leucovorin (5-FU/LV). The tumour activated, oral fluoropyrimidine capecitabine demonstrates superior activity and favourable safety compared with the Mayo regimen, while potentially avoiding the complications and inconvenience associated with i.v. regimens. PATIENTS AND METHODS: Ninety-seven patients with previously untreated advanced/MCRC were randomised to receive capecitabine followed by i.v. 5-FU/LV [Mayo Clinic, in patient de Gramont (IPdG) or out-patient modified de Gramont (OPdG) regimens], or i.v. 5-FU/LV followed by capecitabine. RESULTS: Before treatment, of those patients for whom a preference was recorded, almost all (95%) preferred oral treatment (consistent across all treatment groups) and the majority retained this preference after treatment (64% overall; 86%, 63% and 50% in the Mayo, IPdG and OPdG groups, respectively). Following treatment, the principal reasons for oral treatment preference were increased convenience, home-based administration and tablet formulation. Treatment satisfaction was significantly higher with capecitabine compared with Mayo (P<0.05) and with OPdG compared with capecitabine (P<0.05). Quality of life (QoL) was largely constant across the regimens, although it appeared better with OPdG than capecitabine (P<0.05). Grade 3/4 adverse events were uncommon in all arms. CONCLUSIONS: This study confirmed that the majority of patients with MCRC prefer oral to i.v. therapy, although the OPdG regimen appears to be the most popular i.v. option. Capecitabine clearly represents an effective, well-tolerated oral alternative to i.v. 5-FU/LV. PMID- 16344279 TI - Exploring the relationship between fear of falling and neuroticism: a cross sectional study in community-dwelling women over 70. AB - BACKGROUND: Fear of falling in older adults has been associated with generalised anxiety and may lead to avoidance of activities, with a further negative impact on future falls. Individual differences in personality associated with anxiety have not been previously examined in relation to fear of falling. Current assessment measures and interventions designed to reduce fear of falling in older adults do not take into account perceptions of anxiety associated with individual differences in personality. AIM: To determine whether the core personality trait dimension of neuroticism can predict fear of falling in a community-dwelling sample of women > or = 70 years of age. METHODS: Cross-sectional data from 1,691 UK, community-dwelling female participants aged > or = 70 years were examined using multiple and logistic regression analysis. Fear of falling was measured on a 6-point Likert scale. Neuroticism was measured using the Eysenck personality inventory. RESULTS: The significant independent odds ratios (OR) of predicting fear of falling were: neuroticism (OR 1.47 per SD increase, P<0.001), history of falling (OR 1.57, P<0.001), experience of fracture (OR 1.78, P = 0.014), need to use both arms to push up to rise from a chair (OR 1.56, P = 0.001), poor subjective general health, as measured by the SF12 (OR 1.63 per SD decrease, P<0.001) and living alone (OR 1.31, P = 0.031). CONCLUSIONS: Neuroticism seems to be an important psychological factor in the experience of fear of falling in community-dwelling older women. It may be relevant for inclusion in current assessment measures and for consideration in the design of interventions to reduce fear of falling. PMID- 16344280 TI - Survival ensembles. AB - We propose a unified and flexible framework for ensemble learning in the presence of censoring. For right-censored data, we introduce a random forest algorithm and a generic gradient boosting algorithm for the construction of prognostic and diagnostic models. The methodology is utilized for predicting the survival time of patients suffering from acute myeloid leukemia based on clinical and genetic covariates. Furthermore, we compare the diagnostic capabilities of the proposed censored data random forest and boosting methods, applied to the recurrence-free survival time of node-positive breast cancer patients, with previously published findings. PMID- 16344283 TI - Advocacy for mental health: roles for consumer and family organizations and governments. AB - The World Health Organization urges countries to become more active in advocacy efforts to put mental health on governments' agendas. Health policy makers, planners and managers, advocacy groups, consumer and family organizations, through their different roles and actions, can move the mental health agenda forward. This paper outlines the importance of the advocacy movement, describes some of the roles and functions of the different groups and identifies some specific actions that can be adopted by Ministries of Health. The mental health advocacy movement has developed over the last 30 years as a means of combating stigma and prejudice against people with mental disorders and improving services. Consumer and family organizations and related NGOs have been able to influence governments on mental health policies and laws and educating the public on social integration of people with mental disorders. Governments can promote the development of a strong mental health advocacy sector without compromising this sector's independence. For instance, they can publish and distribute a directory of mental health advocacy groups, include them in their mental health activities and help fledgling groups become more established. There are also some advocacy functions that government officials can, and indeed, should perform themselves. Officials in the ministry of health can persuade officials in other branches of government to make mental health more of a priority, support advocacy activities with both general health workers and mental health workers and carry out public information campaigns about mental disorders and how to maintain good mental health. In conclusion, the World Health Organization believes mental health advocacy is one of the pillars to improve mental health care and the human rights of people with mental disorders. It is hoped that the recommendations in this article will help government officials and activists to strengthen national advocacy movements. PMID- 16344284 TI - TEM-21 extended-spectrum beta-lactamase in a clinical isolate of Alcaligenes faecalis from a nursing home. PMID- 16344285 TI - Empirical antibiotic monotherapy for febrile neutropenia: systematic review and meta-analysis of randomized controlled trials. AB - OBJECTIVES: Early, empirical broad-spectrum antibiotic treatment is the established practice for febrile neutropenia. Several beta-lactams are accepted for monotherapy. We asked whether patients' outcomes are influenced by the chosen beta-lactam. METHODS: Systematic review and meta-analysis of randomized controlled trials comparing anti-pseudomonal beta-lactams administered as empirical monotherapy for febrile neutropenia, with or without vancomycin. The search included The Cochrane Library, PubMed, Embase, Lilacs databases, bibliography, conference proceedings, trial registries and FDA new drug approvals. Two reviewers independently applied selection criteria, performed quality assessment and extracted the data. Trials assessing the same beta-lactam were pooled using the fixed effect model. Relative risks (RRs) with 95% confidence intervals (CIs) were calculated. The primary outcome assessed was all cause mortality. RESULTS: Thirty-three trials fulfilled inclusion criteria. Cefepime was associated with higher all-cause mortality at 30 days than other beta-lactams (RR 1.44, 95% CI 1.06-1.94, 3123 participants). Carbapenems were associated with fewer treatment modifications, including addition of glycopeptides, than ceftazidime or other comparators. Adverse events were significantly more frequent with carbapenems, specifically pseudomembranous colitis (RR 1.94, 95% CI 1.24-3.04, 2025 participants). All-cause mortality was unaltered. Piperacillin/tazobactam was compared only with cefepime and carbapenems, in six trials. No significant differences were demonstrated with paucity of data for all-cause mortality. CONCLUSIONS: The use of cefepime for febrile neutropenia is associated with increased mortality and should be carefully considered pending further analysis. Empirical use of carbapenems entails fewer treatment modifications, but an increased rate of pseudomembranous colitis. Ceftazidime, piperacillin/tazobactam, imipenem/cilastatin and meropenem appear to be suitable agents for monotherapy. PMID- 16344286 TI - Effects of short interfering RNA against methicillin-resistant Staphylococcus aureus coagulase in vitro and in vivo. AB - OBJECTIVES: The emergence of antibiotic-resistant bacteria such as Staphylococcus aureus calls for inventive research and development strategies. Inhibition of bacterial pathogenesis may be a promising therapeutic approach in this regard. The gene-silencing effect of short interfering RNA (siRNA) is useful for this strategy. We investigated the efficacy of siRNA on the expression of coagulase because it is the one of the most important enzymes in the pathogenesis of methicillin-resistant S. aureus (MRSA) infection. METHODS: We designed and synthesized 21 bp siRNA duplexes against staphylococcal coagulase. RT-PCR was performed to determine whether the siRNAs inhibit the expression of the coagulase mRNA and radio-labelled siRNA was used to confirm transfection to bacteria in vitro. The efficacy of siRNA was determined in a murine model of haematogenous pulmonary infection. RESULTS: RT-PCR showed that siRNAs significantly inhibited the expression of the coagulase mRNA. The coagulase titres in the siRNA and control groups were 8 and 32, respectively. Measurement of incorporated radioactivity indicated that the siRNAs were delivered into the bacteria. In the murine infection model, in control and siRNA groups, 7.64 +/- 0.42 and 6.29 +/- 0.23 log cfu/mL (mean +/- SEM) MRSA were detected, respectively, showing that there was a significant decrease in the number of viable bacteria in the siRNA group (P < 0.05). CONCLUSIONS: The results show that siRNA inhibited both mRNA expression and the activity of MRSA coagulase in vitro. The in vivo results revealed that the siRNA was effective in reducing the bacterial load in a murine model of haematogenous pulmonary infection. Targeting of coagulase with siRNA appears to be a novel strategy for treating MRSA infections. PMID- 16344287 TI - In vitro activity of hexadecylphosphocholine (miltefosine) against metronidazole resistant and -susceptible strains of Trichomonas vaginalis. AB - OBJECTIVES: Trichomonas vaginalis is the causative agent of trichomoniasis, a sexually transmitted disease with worldwide significance. Trichomoniasis can be treated with metronidazole; however, resistant strains of T. vaginalis have been isolated and there is a lack of useful alternative drugs. The aim of the present study was to examine the activity of hexadecylphosphocholine (HePC; miltefosine), a membrane-active alkylphospholipid, that is licensed as an antileishmanial agent against T. vaginalis. METHODS: The efficacy of HePC after 30 min, 1 h, 16 h and 24 h against four different T. vaginalis strains (with varying resistance to metronidazole) was evaluated. RESULTS: It was shown that all isolates, including the metronidazole-resistant strains, were susceptible to HePC, with EC50s of between 8 and 40 microM and EC90s of between 8 and 80 microM depending on time and on the medium used for the experiments. Treatment of trichomonads with HePC resulted in rounding up and, at concentrations of >or=40 microM, in subsequent total lysis of the organisms. CONCLUSIONS: HePC may be a promising new candidate for the treatment of trichomoniasis. PMID- 16344288 TI - Sister chromatid exchange and oxidative DNA damage in paving workers exposed to PAHs. AB - Paving workers are exposed during road paving to several polycyclic aromatic hydrocarbons (PAHs) contained in asphalt fumes. In this study early genotoxic and oxidative effects of exposure to bitumen fumes were evaluated in 19 paving workers and 22 controls. Environmental and biological monitoring of exposure was carried out, measuring, on personal air samples from exposed workers collected during three working days, the concentration of 14 PAHs and urinary OH-pyrene at the end of each of the three working days. Genotoxic effect was evaluated analysing sister chromatid exchange (SCE) frequency and direct-oxidative DNA damage by formamido-pyrimidine-glycosylase (Fpg)-modified comet assay on lymphocytes. Tail moment values from Fpg-enzyme treated cells (TMenz) and from untreated cells (TM) were used as parameters of direct and oxidative DNA damage, respectively. For each subject, the TMenz/TM ratio >2.0 was used to indicate the presence of oxidative damage. DNA damage was also evaluated analysing comet percentage. Personal air samples showed low level of total PAHs (2.843 microg m( 3)) with prevalence of 2-3 ring PAHs (2.693 microg m(-3)). Urinary OH-pyrene after work-shift of the three working days was significantly higher than that found at the beginning of the working week. SCE analysis did not show any difference between two groups while an oxidative DNA damage was found in 37% of exposed with respect to the absence in controls. Comet percentage was significantly higher (P = 0.000 ANOVA) in the exposed than in controls. The results demonstrate the high sensitivity of comet assay to assess early oxidative effects induced by exposure to bitumen fumes at low doses and confirm the suitability of urinary OH-pyrene as a biomarker of PAH exposure. In conclusion the study suggests the use of Fpg-modified comet test as a biomarker of early genotoxic effects and that of urinary OH-pyrene as a biomarker of PAH exposure to furnish indications in terms of characterization, prevention and management of risk in occupational exposure to mixtures of potentially carcinogenic substances. PMID- 16344289 TI - The assessment of titanium dioxide exposure. PMID- 16344290 TI - Risk factors associated with the reporting of musculoskeletal symptoms in workers at a laboratory of clinical pathology. AB - OBJECTIVE: This study was conducted with the participation of 120 workers at a laboratory of clinical pathology, with the objective of determining the association of demographic factors, clinical and occupational case histories, individual characteristics, work-related stress and ergonomic workplace analysis with the following outcomes: musculoskeletal symptoms and absences from work associated with these symptoms reported by a questionnaire. METHODS: Interviews were carried out in order to obtain demographic data, occupational case history and risk factors in workplaces. Data related to musculoskeletal symptoms and absences from work have been registered by means of an adaptation of the Nordic Questionnaire. The variables related to stress at work were based on the questionnaire of the European Foundation for the Improvement of Life and Work Conditions. According to the model prepared by the Finnish Institute of Occupational Health, 120 workers' workstations have been submitted for ergonomic analysis of work. RESULTS: By means of multiple logistic regression, the results showed that the symptoms have been significantly associated with the previous history of rheumatic or orthopaedic disease (OR = 15.4; 95% CI, 1.7-135.7) and with movements and postures at work (OR = 13.5; 95% CI, 3.8-47.9). Absences from work due to musculoskeletal symptoms have been significantly associated with the low level of education (OR = 32.3; 95% CI, 4.9-211.8) and marital status (OR = 37.1; 95% CI, 2.3-593.9). CONCLUSIONS: The symptoms have been significantly associated with a previous history of rheumatic or orthopaedic disease and with inadaequate movements and postures at work. Absences from work due to musculoskeletal symptoms have been significantly associated with the workers' low level of education and marital status. PMID- 16344291 TI - Manikin-based performance evaluation of N95 filtering-facepiece respirators challenged with nanoparticles. AB - Protection of the human respiratory system from exposure to nanoparticles is becoming an emerging issue in occupational hygiene. The potential adverse health effects associated with particles of approximately 1-100 nm are probably greater than submicron or micron-sized particles. The performance of two models of N95 half-facepiece-filtering respirators against nano-sized particles was evaluated at two inhalation flow rates, 30 and 85 l min(-1), following a manikin-based protocol. The aerosol concentration was measured outside and inside the facepiece using the Wide-Range Particle Spectrometer. Sodium chloride particles, conventionally used to certify N-series respirators under NIOSH 42 CFR 84 regulations, were utilized as the challenge aerosol. The targeted particle sizes ranged from 10 to 600 nm, although the standard certification tests are performed with particles of approximately 300 nm, which is assumed to be the most penetrating size. The results indicate that the nanoparticle penetration through a face-sealed N95 respirator may be in excess of the 5% threshold, particularly at high respiratory flow rates. Thus, N95 respirators may not always provide the expected respiratory protection for workers. The highest penetration values representing the poorest respirator protection conditions were observed in the particle diameter range of approximately 30-70 nm. Based on the theoretical simulation, we have concluded that for respirators utilizing mechanical filters, the peak penetration indeed occurs at the particle diameter of approximately 300 nm; however, for pre-charged fiber filters, which are commonly used for N95 respirators, the peak shifts toward nano-sizes. This study has confirmed that the neutralization of particles is a crucial element in evaluating the efficiency of a respirator. The variability of the respirator's performance was determined for both models and both flow rates. The analysis revealed that the coefficient of variation of the penetration ranged from 0.10 to 0.54 for particles of 20-100 nm in diameter. The fraction of N95 respirators for which the performance test at 85 l min(-1) demonstrated excessive (>5%) penetration of nanoparticles was as high as 9/10. The test results obtained in a relatively small (0.096 m(3)) test chamber and in a large (24.3 m(3)) walk-in chamber were found essentially the same, thus, suggesting that laboratory-based evaluations have a good potential to adequately represent the respirator field performance. PMID- 16344292 TI - Standing at the crossroads: which way will dementia services evolve? PMID- 16344293 TI - "Leaving town" versus "taking leave": the case for re-thinking academic leave restrictions. PMID- 16344294 TI - Diabetes in African Americans. AB - African Americans have a high risk for type 2 diabetes. Genetic traits, the prevalence of obesity, and insulin resistance all contribute to the risk of diabetes in the African American community. African Americans have a high rate of diabetic complications, because of poor glycaemic control and racial disparities in health care in the USA. African Americans with diabetes may have an atypical presentation that simulates type 1 diabetes, but then their subsequent clinical course is typical of type 2 diabetes. Culturally sensitive strategies, structured disease management protocols, and the assistance of nurses, diabetic educators, and other health care professionals are effective in improving the outcome of diabetes in the African American community. PMID- 16344295 TI - Myocardial infarction in young adults. AB - Although myocardial infarction (MI) mainly occurs in patients older than 45, young men or women can suffer MI. Fortunately, its incidence is not common in patients younger than 45 years. However, the disease carries a significant morbidity, psychological effects, and financial constraints for the person and the family when it occurs at a young age. The causes of MI among patients aged less than 45 can be divided into four groups: (1) atheromatous coronary artery disease; (2) non-atheromatous coronary artery disease; (2) hyper-coagulable states; (4) MI related to substance misuse. There is a considerable overlap between all the groups. This article reviews the literature and highlights the practical issues involved in the management of young adults with MI. PMID- 16344296 TI - Helper T cells and atherosclerosis: the cytokine web. AB - There is growing evidence regarding the importance of inflammation in the pathogenesis of atherosclerosis and its ultimate progression to the clinical syndromes. Recently there has been an increasing interest in the role of helper T (Th) cells in atherosclerosis. The Th cells act with the macrophages and the dendritic cells via the various cytokines in bringing about a variety of changes thus leading to the progression of atherosclerosis. Atherosclerotic lesions have been seen to have increased expression of type 1 helper T (TH1) cells together with increased levels of the Th1 related cytokines. It is mainly the cytokines involved with Th1 functioning that seem to show a prominent effect, with the whole process centred around interferon gamma, making it seem like every pathway and the cytokines involved lead to a final common pathway of interferon gamma secretion; the increase or decrease of which dictates the progression of atherosclerosis and its final manifestation as the clinical syndromes. PMID- 16344297 TI - Unexpected development of artistic talents. AB - The development of exceptional and unexpected artistic skills at any age must be a matter of curiosity. This can occur among young children with severe learning difficulties, especially if they are autistic. Some examples of these so called idiot-savants are given, and the way in which their brains may function. It is also true that elderly people who suffer from frontotemporal dementia can find that they are able to express themselves in remarkable art forms. This can occur in other types of dementia, but then more often it is the changes that result in the paintings of established artists, for example in the paintings of de Kooning. Possible links between these two phenomenon are discussed, and it is suggested that in both instances it may be that if the brain is relieved of a number of functions it can concentrate on the remaining ones. Ways in which this may operate in both groups are reviewed. PMID- 16344298 TI - Differential diagnosis of common tremor syndromes. AB - Tremor is one of the most common involuntary movement disorders seen in clinical practice. In addition to the detailed history, the differential diagnosis is mainly clinical based on the distinction at rest, postural and intention, activation condition, frequency, and topographical distribution. The causes of tremor are heterogeneous and it can present alone (for example, essential tremor) or as a part of a neurological syndrome (for example, multiple sclerosis). Essential tremor and the tremor of Parkinson's disease are the most common tremors encountered in clinical practice. This article focuses on a practical approach to these different forms of tremor and how to distinguish them clinically. Evidence supporting various strategies used in the differentiation is then presented, followed by a review of formal guidelines or recommendations when they exist. PMID- 16344299 TI - What can the Third World learn from the health improvements of Victorian Britain? PMID- 16344300 TI - Liver transplantation for hepatitis C virus related liver disease. AB - Liver transplantation is a useful treatment for end stage liver disease of all aetiologies but recurrent disease presents an ongoing challenge, particularly for hepatitis C virus (HCV) where recurrence is almost universal. Immunosuppression is needed for all patients after transplantation and should be tailored to the individual patient, with particular problems being noted for those with HCV. The longer term effects of immunosuppression, particularly renal failure and the adverse effects of certain treatments on the liver graft, have become more important as survival improves and results are studied for longer periods after transplantation. PMID- 16344301 TI - Diagnosis and treatment of caecal volvulus. AB - Caecal volvulus is an infrequently encountered clinical condition and an uncommon cause of intestinal obstruction. Patients with this condition may present with highly variable clinical presentations ranging from intermittent, self limiting abdominal pain to acute abdominal pain associated with intestinal strangulation and sepsis. Lack of familiarity with this condition is a factor contributing to diagnostic and treatment delays. The objective of this review is to promote clinicians' awareness of this disease through patient case illustration, discussion of disease pathogenesis, clinical features, and management strategies. PMID- 16344302 TI - Agreement between community echocardiography and hospital echocardiography in patients suspected of having left ventricular systolic dysfunction. AB - BACKGROUND: Smaller echocardiography machines, when used in hospitals, are accurate for detecting left ventricular dysfunction and valvular disease. This paper assessed the detection of left ventricular dysfunction and of valvular disease in the community setting by a smaller machine. AIMS: To measure the agreement in patients with suspected heart failure between community echocardiography and traditional echocardiography in the hospital in detecting left ventricular dysfunction and significant valve disease. METHODS: Suspected heart failure patients were referred to one of the authors (SJ) for community echocardiography using a Siemens Cypress machine. The patients had a second echocardiogram in the hospital by another sonographer who was blinded to the results of the first echocardiogram. The reports of the two sonographers were assessed for agreement using kappa statistics. RESULTS: 458 patients had a community echocardiogram and 136 agreed to a second echocardiogram in the hospital. There was excellent agreement, kappa = 0.87 (0.06 SE), for the detection of left ventricular dysfunction between community echocardiography and the hospital machine. The detection of significant valvular disease was good, kappa = 0.75 (0.06) between the community echocardiogram and hospital machines. CONCLUSIONS: In suspected heart failure patients, community echocardiography gives comparable results to traditional hospital echocardiography for left ventricular dysfunction detection and for significant valvular disease detection. PMID- 16344303 TI - Predictors of mortality in mechanically ventilated patients. AB - OBJECTIVES: There are scarce data from India validating scoring systems used to predict outcome in patients requiring mechanical ventilation. This study prospectively compared the organ system failure (OSF), the acute physiology and chronic health evaluation (APACHE) II, and the APACHE III, scores on patients requiring mechanical ventilation in the medical intensive care unit, to predict outcome. METHODS: 200 consecutive patients requiring mechanical ventilation in a medical intensive care unit were recruited. OSF, APACHE II, and APCHE III scores were calculated at admission and daily for one week or until discharge in all patients. Other variables recorded include age, sex, diagnosis, oxygen therapy before ventilation, complications on ventilation, duration in hospital before ventilation, duration of ventilation, type of respiratory failure, alveolar arterial oxygen gradient, P/F ratio, use of tracheostomy, time on ventilator before tracheostomy, muscle relaxant used, fluid balance, inotrope support. Logistic regression analysis and area under the curve were computed to determine which variables independently predict outcome. RESULTS: Of the 200 patients, at discharge 143 patients (71.5%) had died. The factors that independently predicted outcome among these patients on mechanical ventilation were the type of respiratory failure (type I) OR = 2.7 (p = 0.02), the use of inotropes OR 2.4 (p = 0.04), and the APACHE II score OR = 1.8 (p = 0.008) for every five point increase in APACHE II score. CONCLUSIONS: Type 1 respiratory failure, the use of inotropes, and the APACHE II score measured at admission are significant independent predictors of mortality in the patients on mechanical ventilation. PMID- 16344305 TI - Dynamic confidence during simulated clinical tasks. AB - OBJECTIVE: Doctors' confidence in their actions is important for clinical performance. While static confidence has been widely studied, no study has examined how confidence changes dynamically during clinical tasks. METHOD: The confidence of novice (n = 10) and experienced (n = 10) trainee anaesthetists was measured during two simulated anaesthetic crises, bradycardia (easy task) and failure to ventilate (difficult task). RESULTS: As expected, confidence was high in the novice and experienced groups in the easy task. What was surprising, however, was that confidence during the difficult task decreased for both groups, despite appropriate performance. CONCLUSIONS: Given that confidence affects performance, it is alarming that doctors who may be acting unsupervised should lose dynamic confidence so quickly. Training is needed to ensure that confidence does not decrease inappropriately during a correctly performed procedure. Whether time on task interacts with incorrect performance to produce further deficits in confidence should now be investigated. PMID- 16344307 TI - Reduced Apaf-1 levels in cardiomyocytes engage strict regulation of apoptosis by endogenous XIAP. AB - Overexpression studies have identified X-linked inhibitor of apoptosis protein (XIAP) as a potent inhibitor of caspases. However, the exact function of endogenous XIAP in regulating mammalian apoptosis is less clear. Endogenous XIAP strictly regulates cytochrome c-dependent caspase activation in sympathetic neurons but not in many mitotic cells. We report that postmitotic cardiomyocytes, unlike fibroblasts, are remarkably resistant to cytosolic microinjection of cytochrome c. The cardiomyocyte resistance to cytochrome c is mediated by endogenous XIAP, as XIAP-deficient cardiomyocytes die rapidly with cytosolic cytochrome c alone. Importantly, we found that cardiomyocytes, like neurons, have markedly reduced Apaf-1 levels and that this decrease in Apaf-1 is directly linked to the tight regulation of caspase activation by XIAP. These data identify an important function of XIAP in cardiomyocytes and point to a striking similarity in the regulation of apoptosis in postmitotic cells. PMID- 16344308 TI - The mammalian Scribble polarity protein regulates epithelial cell adhesion and migration through E-cadherin. AB - Scribble (Scrib) is a conserved polarity protein required in Drosophila melanogaster for synaptic function, neuroblast differentiation, and epithelial polarization. It is also a tumor suppressor. In rodents, Scrib has been implicated in receptor recycling and planar polarity but not in apical/basal polarity. We now show that knockdown of Scrib disrupts adhesion between Madin Darby canine kidney epithelial cells. As a consequence, the cells acquire a mesenchymal appearance, migrate more rapidly, and lose directionality. Although tight junction assembly is delayed, confluent monolayers remain polarized. These effects are independent of Rac activation or Scrib binding to betaPIX. Rather, Scrib depletion disrupts E-cadherin-mediated cell-cell adhesion. The changes in morphology and migration are phenocopied by E-cadherin knockdown. Adhesion is partially rescued by expression of an E-cadherin-alpha-catenin fusion protein but not by E-cadherin-green fluorescent protein. These results suggest that Scrib stabilizes the coupling between E-cadherin and the catenins and are consistent with the idea that mammalian Scrib could behave as a tumor suppressor by regulating epithelial cell adhesion and migration. PMID- 16344309 TI - Ubiquitin/SUMO modification of PCNA promotes replication fork progression in Xenopus laevis egg extracts. AB - The homotrimeric DNA replication protein proliferating cell nuclear antigen (PCNA) is regulated by both ubiquitylation and sumoylation. We study the appearance and the impact of these modifications on chromosomal replication in frog egg extracts. Xenopus laevis PCNA is modified on lysine 164 by sumoylation, monoubiquitylation, and diubiquitylation. Sumoylation and monoubiquitylation occur during the replication of undamaged DNA, whereas diubiquitylation occurs specifically in response to DNA damage. When lysine 164 modification is prevented, replication fork movement through undamaged DNA slows down and DNA polymerase delta fails to associate with replicating chromatin. When sumoylation alone is prevented, replication occurs normally and neither monoubiquitylation nor sumoylation are required for the replication of simple single-strand DNA templates. Our findings expand the repertoire of functions for PCNA ubiquitylation and sumoylation by elucidating a role for these modifications during the replication of undamaged DNA. Furthermore, they suggest that PCNA monoubiquitylation serves as a molecular gas pedal that controls the speed of replisome movement during S phase. PMID- 16344310 TI - Mutational analyses reveal a novel function of the nucleotide-binding domain of gamma-tubulin in the regulation of basal body biogenesis. AB - We have used in vitro mutagenesis and gene replacement to study the function of the nucleotide-binding domain (NBD) of gamma-tubulin in Tetrahymena thermophila. In this study, we show that the NBD has an essential function and that point mutations in two conserved residues lead to over-production and mislocalization of basal body (BB) assembly. These results, coupled with previous studies (Dammermann, A., T. Muller-Reichert, L. Pelletier, B. Habermann, A. Desai, and K. Oegema. 2004. Dev. Cell. 7:815-829; La Terra, S., C.N. English, P. Hergert, B.F. McEwen, G. Sluder, and A. Khodjakov. 2005. J. Cell Biol. 168:713-722), suggest that to achieve the precise temporal and spatial regulation of BB/centriole assembly, the initiation activity of gamma-tubulin is normally suppressed by a negative regulatory mechanism that acts through its NBD. PMID- 16344311 TI - Regulation of Sar1 NH2 terminus by GTP binding and hydrolysis promotes membrane deformation to control COPII vesicle fission. AB - The mechanisms by which the coat complex II (COPII) coat mediates membrane deformation and vesicle fission are unknown. Sar1 is a structural component of the membrane-binding inner layer of COPII (Bi, X., R.A. Corpina, and J. Goldberg. 2002. Nature. 419:271-277). Using model liposomes we found that Sar1 uses GTP regulated exposure of its NH2-terminal tail, an amphipathic peptide domain, to bind, deform, constrict, and destabilize membranes. Although Sar1 activation leads to constriction of endoplasmic reticulum (ER) membranes, progression to effective vesicle fission requires a functional Sar1 NH2 terminus and guanosine triphosphate (GTP) hydrolysis. Inhibition of Sar1 GTP hydrolysis, which stabilizes Sar1 membrane binding, resulted in the formation of coated COPII vesicles that fail to detach from the ER. Thus Sar1-mediated GTP binding and hydrolysis regulates the NH2-terminal tail to perturb membrane packing, promote membrane deformation, and control vesicle fission. PMID- 16344312 TI - Gemcitabine doublets in advanced pancreatic cancer: should we move on? PMID- 16344313 TI - Axillary surgery: clinical judgment required. PMID- 16344314 TI - Gene expression profiling of localized esophageal carcinomas: association with pathologic response to preoperative chemoradiation. AB - PURPOSE: Patients with localized esophageal carcinoma have a 5-year survival rate of less than 20%. Patients are often treated similarly (ie, with preoperative chemoradiotherapy) but the outcomes vary greatly. Chemoradiotherapy and surgery can result in significant undesirable consequences. Currently, however, there are no tools to help select optimum therapy. We hypothesized that gene expression profiling could provide clues and biomarkers for selection of therapy. METHODS: Pretreatment endoscopic cancer biopsies from 19 patients (16 with adenocarcinoma, two with squamous cell carcinoma, and one with adenosquamous carcinoma) enrolled onto a preoperative chemoradiotherapy protocol were profiled using oligonucleotide microarrays. Surgical specimens following therapy were assessed for the degree of pathologic response. On the basis of array data, selected genes were analyzed by polymerase chain reaction. RESULTS: Unsupervised hierarchical cluster analysis segregated the cancers into two molecular subtypes, each consisting 10 and nine specimens, respectively. Most cancers (five of six) that had pathologic complete response (pathCR) clustered in molecular subtype I. Subtype II, with one exception, consisted cancers that had less than pathCR (< pathCR). Using a combination marker approach, levels of PERP, S100A2, and SPRR3 allowed discrimination of pathCR from < pathCR with high sensitivity and specificity (85%). Pathway analysis identified apoptotic pathway as one of the key functions downregulated in molecular type II in comparison with type I. CONCLUSION: These encouraging, albeit preliminary, data suggest that expression profiling may distinguish cancers with different pathologic outcome. This is the first report to show subtypes of esophageal cancers with distinct molecular signatures. The potential of PERP, S100A2, and SPRR3 as biomarkers of pathCR warrants further validation. PMID- 16344315 TI - High complete remission rate and promising outcome by combination of imatinib and chemotherapy for newly diagnosed BCR-ABL-positive acute lymphoblastic leukemia: a phase II study by the Japan Adult Leukemia Study Group. AB - PURPOSE: A novel therapeutic approach is urgently needed for BCR-ABL-positive acute lymphoblastic leukemia (ALL). In this study, we assessed the efficacy and feasibility of chemotherapy combined with imatinib. PATIENTS AND METHODS: A phase II study of imatinib-combined chemotherapy was conducted for newly diagnosed BCR ABL-positive ALL in adults. Eighty patients were entered into the trial between September 2002 and January 2005. RESULTS: Remission induction therapy resulted in complete remission (CR) in 77 patients (96.2%), resistant disease in one patient, and early death in two patients, as well as polymerase chain reaction negativity of bone marrow in 71.3%. The profile and incidence of severe toxicity were not different from those associated with our historic chemotherapy-alone regimen. Relapse occurred in 20 patients after median CR duration of 5.2 months. Allogeneic hematopoietic stem-cell transplantation (HSCT) was performed for 49 patients, 39 of whom underwent transplantation during their first CR. The 1-year event-free and overall survival (OS) rates were estimated to be 60.0%, and 76.1%, respectively, which were significantly better than those for our historic controls treated with chemotherapy alone (P < .0001 for both). Among the current trial patients, the probability for OS at 1 year was 73.3% for those who underwent allogeneic HSCT, and 84.8% for those who did not. CONCLUSION: Our results demonstrated that imatinib-combined regimen is effective and feasible for newly diagnosed BCR-ABL-positive ALL. Despite a relatively short period of observation, a major potential of this treatment is recognized. Longer follow-up is required to determine its overall effect on survival. PMID- 16344316 TI - Treatment for acute myelogenous leukemia by low-dose, total-body, irradiation based conditioning and hematopoietic cell transplantation from related and unrelated donors. AB - PURPOSE: The use of low-dose, irradiation-based preparative regimens have allowed the extension of allografting to older and medically infirm patients. The study reported here assessed outcomes for patients with acute myeloid leukemia (AML) in different stages of their disease, who were not considered candidates for conventional hematopoietic cell transplantation (HCT) because of age and/or other known risk factors and were given minimal conditioning followed by HCT from related or unrelated donors. PATIENTS AND METHODS: The present study included 122 patients with AML, who were conditioned with 2 Gy total-body irradiation (TBI) on day 0 with or without preceding fludarabine (30 mg/m2/d from days -4 to -2), and given postgrafting cyclosporine at 6.25 mg/kg twice daily from day -3 and mycophenolate mofetil at 15 mg/kg twice daily from day 0. RESULTS: Durable engraftment was observed in 95% of the patients. Cumulative incidences of acute graft-versus-host disease grades 2 to 4 at 6 months were 35% after related and 42% after unrelated HCT, respectively. With a median follow-up of 44 months (range, 26 to 79 months), 51 patients were alive, of whom 48 were in complete remission (CR). Cumulative nonrelapse mortalities were 10% and 22%, and cumulative mortalities from disease progression were 47% and 33% at 2 years for related and unrelated recipients, respectively. Overall, 2-year survival was 48%, and disease-free survival was 44%. Patients receiving transplantation in CR1 had 2-year overall survivals of 44% after related and 63% after unrelated HCT, respectively. CONCLUSION: We conclude that HCT from related and unrelated donors after low-dose TBI is a promising treatment for elderly patients with AML. PMID- 16344317 TI - Select high-risk genetic features predict earlier progression following chemoimmunotherapy with fludarabine and rituximab in chronic lymphocytic leukemia: justification for risk-adapted therapy. AB - PURPOSE: Several new prognostic factors predicting rapid disease progression in chronic lymphocytic leukemia (CLL) have been identified, including unmutated Ig V(H) mutational status, del(11)(q23), del(17)(p13.1), and p53 mutations. To date, the impact of these same prognostic factors have not been examined relative to treatment outcome with chemoimmunotherapy. METHODS: We examined the impact of these new prognostic factors on predicting treatment outcome in symptomatic, untreated CLL patients who received chemoimmunotherapy with fludarabine and rituximab as part of a completed, randomized phase II study, Cancer and Leukemia Group B (CALGB) 9712. RESULTS: Eighty-eight patients treated as part of CALGB 9712 had detailed prognostic factor assessment performed. Using Ig V(H) mutational status to classify risk, there was no association between complete response rate with either unmutated Ig V(H) mutational status or high-risk interphase cytogenetics. However, the median progression-free survival (PFS; P = .048) and overall survival (OS; P = .01) were shorter among the Ig V(H) unmutated patients as compared with the Ig V(H) mutated patients. Using the hierarchical classification of Dohner, PFS (P = .005) and OS (P = .004) were significantly longer as the classification moved from high risk [del (11)(q22.3) or del (17)(p13.1)] to low risk. CONCLUSION: These data demonstrate that high-risk CLL patients characterized by Ig V(H) unmutated (> or = 98%) or high-risk interphase cytogenetics, including either del(17p) or del(11q), appear to have a shorter PFS and OS with chemoimmunotherapy. Larger prospective studies will be required to determine the independent influence of Ig V(H) mutational status and interphase cytogenetics on treatment outcome. PMID- 16344318 TI - New MUC1 serum immunoassay differentiates pancreatic cancer from pancreatitis. AB - PURPOSE: To evaluate a new immunoassay for identification and quantitation of MUC1 in the sera of patients with pancreatic cancer or pancreatitis. The sensitivity and specificity of the assay are examined and compared to results from a CA19-9 immunoassay. METHODS: An in vitro enzyme immunoassay was established with monoclonal antibody PAM4 as the capture reagent, and a polyclonal anti-MUC1 antibody as the probe. Patient sera were obtained from healthy, adult patients with acute and chronic pancreatitis, and those with pancreatic and other forms of cancer, and were measured for PAM4-reactive MUC1. RESULTS: At a cutoff of 10.2 units/mL, 41 (77%) of 53 pancreatic cancer patients, none of the healthy individuals (n = 43), and only four (5%) of 87 patients with pancreatitis were positive above this value. Among nonpancreatic cancers investigated, colorectal cancers gave the highest percentage of positives (14%; five of 36). Overall, the sensitivity and specificity of the immunoassay for pancreatic cancer were 77% and 95%, respectively. Receiver operator characteristic analyses for discrimination of pancreatic cancer from pancreatitis provided an area under the curve of 0.89 (95% CI, 0.82 to 0.93), with a specificity of 95.4% and a positive likelihood ratio of 16.8. A direct pair-wise comparison of PAM4 and CA19-9 immunoassays for discrimination of pancreatic cancer and pancreatitis resulted in a significant difference (P < .003), with the PAM4 immunoassay demonstrating superior sensitivity and specificity. CONCLUSION The high sensitivity and specificity observed suggest that the PAM4-based immunoassay of circulating MUC1 may be useful in the diagnosis of pancreatic cancer. PMID- 16344319 TI - High frequency of submicroscopic hemizygous deletion is a major mechanism of loss of expression of PTEN in uveal melanoma. AB - PURPOSE: Although cytogenetic aberrations at 10q have been reported in up to 27% of uveal melanomas, the role of PTEN in the pathogenesis of uveal melanoma is largely unknown. Our aim was to determine the frequency and clinical significance of PTEN alterations in uveal melanomas. PATIENTS AND METHODS: We examined PTEN expression using immunohistochemistry in 75 sporadic uveal melanomas, with an average follow-up of 89 months. Molecular cytogenetic alterations were studied using comparative genomic hybridization (CGH). Genotyping was carried out using an intragenic PTEN marker and two flanking markers. Mutational analysis of PTEN was also carried out. RESULTS: Of the 75 tumors, 12 (16%) showed no PTEN immunostaining, 32 (42.7%) showed weak to moderate staining and the remaining 31 (38.2%) showed staining similar to the normal internal controls. Using CGH, only two (15.4%) of 13 samples showed any loss of 10q. However, in the 38 tumors with informative genotyping, we found that 29 (76.3%) had loss of heterozygosity (LOH) of at least one PTEN marker, and 15 (39.5%) showed LOH of at least two markers. Mutations in the coding region of PTEN were identified in four (11.4%) of 35 tumors. Further, loss of cytoplasmic PTEN expression by immunohistochemistry was associated with shortened disease-free survival (P = .029). CONCLUSION: This is the first demonstration that PTEN is a tumor suppressor involved in uveal melanoma pathogenesis and may be associated with clinical outcome. Our data also suggest that submicroscopic deletion, but not large deletions, is the major mechanism of loss of PTEN expression in uveal melanomas. PMID- 16344320 TI - Phase II study of fixed dose rate gemcitabine with cisplatin for metastatic adenocarcinoma of the pancreas. AB - PURPOSE: Although gemcitabine remains the standard of care for patients with advanced pancreatic cancer, additional improvements may be realized by combining therapeutic agents with synergistic activity, and optimizing drug delivery using pharmacokinetic principles such as fixed dose rate (FDR) infusion. The objectives of this study were to determine safety and efficacy in patients with metastatic pancreatic cancer treated with FDR gemcitabine in combination with low-dose cisplatin. PATIENTS AND METHODS: Chemotherapy-naive patients with metastatic pancreatic adenocarcinoma were treated with a combination of gemcitabine 1,000 mg/m2 at 10 mg/m2/min together with cisplatin 20 mg/m2 on days 1 and 8 of a 21 day cycle. Patient follow-up was performed using computerized tomographic scans and serial CA 19-9 measurements. RESULTS: A total of 51 patients were enrolled onto the study, with a median follow-up time of 215 days. Twenty-two of 40 patients (55.0%) with a baseline serum CA 19-9 level > or = 2x the upper limit of normal demonstrated a > or = 50% biomarker decline during treatment. Nine of 47 patients (19.1%) with measurable disease achieved a partial response, and 28 patients (59.6%) had disease stabilization for at least two treatment cycles. Median time to progression was 3.9 months and median survival was 7.1 months, with an estimated 1-year survival rate of 29%. The most frequently reported grade 3 or 4 adverse events were neutropenia (52.9%) and thrombocytopenia (15.7%). Most patients were switched to an every-other-week dosing schedule. CONCLUSION: The combination of FDR gemcitabine and cisplatin is well tolerated and appears to be an acceptable, albeit not clearly superior, alternative to other gemcitabine/platinum regimens for the treatment of metastatic pancreatic cancer. PMID- 16344321 TI - Randomized trial comparing axillary clearance versus no axillary clearance in older patients with breast cancer: first results of International Breast Cancer Study Group Trial 10-93. AB - PURPOSE: Axillary clearance in early breast cancer aims to improve locoregional control and provide staging information but is associated with undesirable morbidity. We therefore investigated whether avoiding axillary surgery in older women would result in improved quality of life (QL) with similar disease-free survival (DFS) and overall survival (OS). PATIENTS AND METHODS: Between 1993 and 2002, women > or = 60 years old with clinically node-negative operable breast cancer in whom adjuvant tamoxifen was considered indicated regardless of pathologic nodal status were randomly assigned to primary surgery plus axillary clearance (Sx + Ax) followed by tamoxifen (Tam) versus Sx without Ax followed by Tam for 5 consecutive years. The primary end point was QL reported by the patient and by physician assessment. RESULTS: A total of 473 patients (234 to Sx + Ax, 239 to Sx) were randomly assigned. The median age was 74 years; 80% had estrogen receptor-positive disease. In both the patients' subjective assessment of their QL and the physicians' perception of the patients' QL, the largest adverse QL effects of Ax were observed from baseline to the first postoperative assessment, but the differences tended to disappear in 6 to 12 months. At a median follow-up of 6.6 years, results for Sx + Ax and Sx yielded similar DFS (6-year DFS, 67% v 66%; hazard ratio [HR] Sx + Ax/Sx, 1.06; 95% CI, 0.79 to 1.42; P = .69) and OS (6 year OS, 75% v 73%; HR Sx + Ax/Sx, 1.05; 95% CI, 0.76 to 1.46; P = .77). CONCLUSION: Avoiding axillary clearance for women > or = 60 years old who have clinically node-negative disease and receive Tam for endocrine-responsive disease yields similar efficacy with better early QL. PMID- 16344322 TI - Vascular endothelial growth factor-D and its receptor VEGFR-3: two novel independent prognostic markers in gastric adenocarcinoma. AB - PURPOSE: Vascular endothelial growth factor (VEGF)-D and its homolog VEGF-C influence lymphangiogenesis through activation of VEGF receptor 3 (VEGFR-3), and have been implicated in lymphatic tumor spread. Nodal dissemination of gastric adenocarcinomas critically determines clinical outcome and therapeutic options of affected patients. Therefore, we analyzed expression and prognostic significance of VEGF-D along with VEGF-C, and VEGFR-3 in gastric adenocarcinomas. MATERIALS AND METHODS: VEGF-C, VEGF-D, and VEGFR-3 were analyzed in 91 R(0)-resected primary gastric adenocarcinomas, corresponding noncancerous gastric mucosa, and lymph node metastases employing immunohistochemistry and/or in situ hybridization. Blood and lymph vessel densities were assessed after staining with CD31 and LYVE-1-specific antibodies. RESULTS: VEGF-D and VEGF-C were detected in 67.0% and 50.5% of gastric cancers, respectively. Healthy gastric mucosa was negative for VEGF-C and in 12.5% positive for VEGF-D. Presence of VEGF-D (P = .005) or VEGF-C (P = .006) was correlated with lymphatic metastases and decreased survival (VEGF-D, P < .05; VEGF-C, P < .05). VEGFR-3 was correlated with reduced carcinoma-specific survival (P < .05), and Cox multivariate regression analysis qualified VEGF-D and VEGFR-3, but not VEGF-C, as independent prognostic parameters. In lymph node-positive gastric cancers, presence of VEGF-D/VEGFR-3 was associated with poor survival, whereas absence of VEGF-D/VEGFR-3 defined a subgroup of patients with clearly favorable prognosis. CONCLUSION: VEGF-D and VEGFR-3 are novel independent prognostic marker molecules aiding to identify patients with poor prognosis after curative resection of gastric adenocarcinomas. Combined analysis of the VEGF-C/VEGF-D/VEGFR-3 system can be useful to identify patients with unfavorable clinical outcome and thereby may help to refine therapeutic decisions in gastric cancer. PMID- 16344324 TI - Improved Golgi-like visualization in retrogradely projecting neurons after EGFP adenovirus infection in adult rat and monkey. AB - An adenovirus vector was generated using a neuron-specific promoter synapsin I and enhanced green fluorescent protein (EGFP) reporter (AdSynEGFP). In addition, two modifications were identified that resulted in robust and reliable retrograde transport and EGFP expression after injection of the virus into three different brain regions in adult rats (medial prefrontal cortex, posterior thalamic nuclear group, and CA1). These are postinjection survival times of 14 days and addition of high concentrations of NaCl (>or=600 mM) to the injection buffer. These modifications resulted in obvious improvement in the intensity of the EGFP signal and in the number of labeled cells. Use of anti-EGFP in immunofluorescence or immunoperoxidase processing further enhanced the signal so that Golgi-like filling of dendritic spines and axon collaterals was routinely achieved. Effectiveness of the AdSynEGFP for Golgi-like filling was confirmed in one rhesus monkey with injections in visual area V4. Because of the long-term viability of the infected neurons (at least up to 28 days in rats and 22 days in monkey), this AdSynEGFP is suitable for use in microcircuitry studies in combination with other fluorescently tagged elements, including anterogradely labeled extrinsic projections. The native EGFP signal (without antibody enhancement) may be sufficient for studies involving cultured cells or slices. PMID- 16344323 TI - Effects of multiple metal binding sites on calcium and magnesium-dependent activation of BK channels. AB - BK channels are activated by physiological concentrations of intracellular Ca2+ and Mg2+ in a variety of cells. Previous studies have identified two sites important for high-affinity Ca2+ sensing between [Ca2+]i of 0.1-100 microM and a site important for Mg2+ sensing between [Mg2+]i of 0.1-10 mM. BK channels can be also activated by Ca2+ and Mg2+ at concentrations>10 mM so that the steady-state conductance and voltage (G-V) relation continuously shifts to more negative voltage ranges when [Mg2+]i increases from 0.1-100 mM. We demonstrate that a novel site is responsible for metal sensing at concentrations>=10 mM, and all four sites affect channel activation independently. As a result, the contributions of these sites to channel activation are complex, depending on the combination of Ca2+ and Mg2+ concentrations. Here we examined the effects of each of these sites on Ca2+ and Mg2+-dependent activation and the data are consistent with the suggestion that these sites are responsible for metal binding. We provide an allosteric model for quantitative estimation of the contributions that each of these putative binding sites makes to channel activation at any [Ca2+]i and [Mg2+]i. PMID- 16344325 TI - Signet ring cells in gastric carcinomas are derived from neuroendocrine cells. AB - Adenocarcinomas are malignant tumors with glandular growth and/or supposed intracellular mucin as identified by periodic acid-Schiff (PAS) positivity. Gastric signet ring cell carcinomas are classified as diffuse type. A proportion of diffuse-type adenocarcinomas have previously been suggested to be of neuroendocrine origin. In the present study we examined gastric signet ring cell carcinomas for neuroendocrine differentiation. Of 11 gastric signet ring cell carcinomas, 8 contained areas with PAS-positive signet ring cells that also were immunoreactive for one or several neuroendocrine markers: synaptophysin, chromogranin A, and histidine decarboxylase, the latter an enterochromaffin-like (ECL) cell marker. Whereas PAS positivity was located in the central cytoplasm, neuroendocrine immunoreactivity was often located as a rim surrounding an otherwise non-immunoreactive cytoplasm, presumed to represent the area with PAS positive material. These findings indicate that signet ring cell carcinomas could be of neuroendocrine origin. We propose that signet ring cell carcinomas develop by gradual dedifferentiation from ECL cells via signet ring cells with neuroendocrine immunoreactivity toward signet ring cells where the cytoplasm mainly consists of PAS-positive material. This finding could have implications for the classification and understanding of gastric carcinogenesis. PMID- 16344326 TI - Intrastriatal infusion of liver growth factor stimulates dopamine terminal sprouting and partially restores motor function in 6-hydroxydopamine-lesioned rats. AB - Liver growth factor (LGF) is a mitogen for liver cells that shows biological activity in extrahepatic sites and may be useful for neuroregenerative therapies. The aim of this work was to investigate the effects of the intrastriatal (IS) infusion of LGF in the 6-hydroxydopamine rat model of Parkinson's disease. Tyrosine hydroxylase-positive innervation was significantly increased in the dopamine-denervated striatum of rats receiving intrastriatal LGF infusions (160 ng/day/rat x 15 days) as compared with a vehicle-infused group. There was no evidence of dopaminergic neurogenesis in the striatum or substantia nigra in any experimental group at the times studied. However, in those animals undergoing IS LGF infusion for 48 hr, we found a significant increase in both microglial proliferation and in the number of microglial cells that acquired the ameboid morphology. This is characteristic of activated microglia/macrophages that has been reported to play an important role in dopamine terminal sprouting. In summary, our study shows that IS infusion of LGF stimulates the outgrowth of tyrosine hydroxylase-positive terminals in the striatum of 6-hydroxydopamine treated rats. As apomorphine-induced rotational behavior was also reduced in these animals, we propose LGF as a novel factor that, when delivered to the striatum, may be useful in the treatment of Parkinson's disease. PMID- 16344327 TI - Mast cell contributes to cardiomyocyte apoptosis after coronary microembolization. AB - Coronary microembolization (CME) is associated with progressive myocardial dysfunction despite restoration of coronary flow reserve (CFR). The potential pathophysiological role of mast cells (MCs) remains unclear. Therefore, we induced CME in 18 miniswines and determined whether MC accumulation occurs and their effects on local cytokine secretion [interleukin (IL)-6, IL-8, tumor necrosis factor-alpha (TNF-alpha)]; cardiomyocyte apoptosis; and collagen formation at day 1 (D1), day 7 (D7), and day 30 (D30) after CME. Four sham operated animals without CME (controls) and six animals treated with a MC stabilization agent (tranilast) for 30 days after CME were also studied. CFR decreased at D1 but returned to baseline level at D7 and D30. Coronary sinus levels of IL-6, IL-8, and TNF-alpha increased significantly at D1 and D7 (p<0.01 vs baseline). Levels of IL-6 and IL-8 at D30 returned to baseline level, but not those of TNF-alpha. The numbers of total and degranulating MCs, % apoptotic cardiomyocytes, and collagen volume fraction (CVF) over CME myocardium at D1, D7, and D30 were significantly higher than controls (p<0.01). Treatment with tranilast significantly reduced the serum level of TNF-alpha, numbers of total and degranulating MCs, % apoptotic cardiomyocytes, and CVF at D30 (all p<0.05). There was a significant positive correlation between the numbers of MCs with % apoptotic cardiomyocytes (r = 0.77, p<0.001) and CVF (r = 0.75, p<0.001) over the CME myocardium. Despite restoration of CFR, cardiomyocyte apoptosis persisted after CME and was positively correlated with the number of MCs but was prevented with tranilast treatment. These findings suggest that MCs contribute to cardiomyocyte apoptosis after CME. PMID- 16344328 TI - Assessing the expression of enterotoxigenic Escherichia coli-specific surface antigens in recombinant strains by transmission electron microscopy and immunolabeling. AB - Infections with enterotoxigenic Escherichia coli (ETEC) are a major cause of travelers' diarrhea worldwide. Colonization of the small intestine mucosa is dependent on specific colonization factor antigens (CFA) and coli surface (CS) antigens. CFA/1, CS3, and CS6 are the most prevalent fimbrial antigens found in clinical isolates. The goal of our study was to visualize the morphology of CS3 and CS6 fimbriae in wild-type and recombinant E. coli strains by means of transmission electron microscopy in conjunction with negative staining and immunolabeling. Corresponding ETEC genes were cloned into E. coli K12 strain DH10B. Expression of fimbriae was dependent on culture conditions and sample handling. Specific immunolabeling of fimbriae unequivocally demonstrated the presence of all types of surface antigens investigated. Negative staining was effective in revealing CS3 but not CS6. In addition, this technique clearly demonstrated differences in the morphology of genetically and immunologically identical CS3 surface antigens in wild-type and recombinant strains. This paper provides a basis for the assessment of recombinant vaccines. PMID- 16344329 TI - Expression of tandem P domain K+ channel, TREK-1, in the rat carotid body. AB - TREK-1 is one of the important potassium channels for regulating membrane excitability. To examine the distribution of TREK-1 in the rat carotid body, we performed RT-PCR for mRNA expression and in situ hybridization and immunohistochemistry for tissue distribution of TREK-1. RT-PCR detected mRNA expression of TREK-1 in the carotid body. Furthermore, in situ hybridization revealed the localization of TREK-1 mRNA in the glomus cells. TREK-1 immunoreactivity was mainly distributed in the glomus cells and nerve fibers in the carotid body. TREK-1 may modulate potassium current of glomus cells and/or afferent nerve endings in the rat carotid body. PMID- 16344330 TI - Fibromodulin-deficient mice display impaired collagen fibrillogenesis in predentin as well as altered dentin mineralization and enamel formation. AB - To determine the functions of fibromodulin (Fmod), a small leucine-rich keratan sulfate proteoglycan in tooth formation, we investigated the distribution of Fmod in dental tissues by immunohistochemistry and characterized the dental phenotype of 1-day-old Fmod-deficient mice using light and transmission electron microscopy. Immunohistochemistry was also used to compare the relative protein expression of dentin sialoprotein (DSP), dentin matrix protein-1 (DMP 1), bone sialoprotein (BSP), and osteopontin (OPN) between Fmod-deficient mice and wild type mice. In normal mice and rats, Fmod immunostaining was mostly detected in the distal cell bodies of odontoblasts and in the stratum intermedium and was weaker in odontoblast processes and predentin. The absence of Fmod impaired dentin mineralization, increased the diameter of the collagen fibrils throughout the whole predentin, and delayed enamel formation. Immunohistochemistry provides evidence for compensatory mechanisms in Fmod-deficient mice. Staining for DSP and OPN was decreased in molars, whereas DMP 1 and BSP were enhanced. In the incisors, labeling for DSP, DMP 1, and BSP was strongly increased in the pulp and odontoblasts, whereas OPN staining was decreased. Positive staining was also seen for DMP 1 and BSP in secretory ameloblasts. Together these studies indicate that Fmod restricts collagen fibrillogenesis in predentin while promoting dentin mineralization and the early stages of enamel formation. PMID- 16344331 TI - Rotigaptide (ZP123) prevents spontaneous ventricular arrhythmias and reduces infarct size during myocardial ischemia/reperfusion injury in open-chest dogs. AB - The antiarrhythmic and cardioprotective effect of increasing gap junction intercellular communication during ischemia/reperfusion injury has not been studied. The antiarrhythmic peptide rotigaptide (previously ZP123), which maintains gap junction intercellular communication, was tested in dogs subjected to a 60-min coronary artery occlusion and 4 h of reperfusion. Rotigaptide was administered i.v. 10 min before reperfusion as a bolus + i.v. infusion at doses of 1 ng/kg bolus + 10 ng/kg/h infusion (n = 6), 10 ng/kg bolus + 100 ng/kg/h infusion (n = 5), 100 ng/kg bolus + 1000 ng/kg/h infusion (n = 8), 1000 ng/kg bolus + 10 mug/kg/h infusion (n = 6), and vehicle control (n = 5). Premature ventricular complexes (PVCs) were quantified during reperfusion. A series of four or more consecutive PVCs was defined as ventricular tachycardia (VT). The total incidence of VT was reduced significantly with the two highest doses of rotigaptide (20.3 +/- 10.9 and 4.3 +/- 4.1 events; p < 0.05) compared with controls (48.7 +/- 6.0). Total PVCs were reduced significantly from 25.1 +/- 4.2% in control animals to 11.0 +/- 4.4 and 1.7 +/- 1.3% after the two highest doses of rotigaptide. Infarct size, expressed as a percentage of the left ventricle, was reduced significantly from 13.2 +/- 1.9 in controls to 7.1 +/- 1.0 (p < 0.05) at the highest dose of rotigaptide. Ultrastructural evaluation revealed no differences in myocardial injury in the infarct area, area at risk, border zone, or normal zone in vehicle and rotigaptide-treated animals. However, rotigaptide did increase the presence of gap junctions in the area at risk (p = 0.022, Fisher's exact test). Rotigaptide had no effect on heart rate, blood pressure, heart rate-corrected QT interval, or left ventricular end-diastolic pressure. In conclusion, these results demonstrate that rotigaptide is a potent antiarrhythmic compound with cardioprotective effects and desirable safety. PMID- 16344333 TI - Corticosteroids prevent myofibroblast accumulation and airway remodeling in mice. AB - At present there are conflicting results from studies investigating the role of corticosteroids in inhibiting airway remodeling in asthma. We have used a mouse model to determine whether administration of corticosteroids prevents the development of allergen-induced structural features of airway remodeling. Mice treated with corticosteroids were subjected to repetitive ovalbumin (OVA) challenge for 3 mo, at which time levels of peribronchial fibrosis and the thickness of the peribronchial smooth muscle layer were assessed by immunohistology, levels of transforming growth factor (TGF)-beta1 by ELISA, and the number of alpha-smooth muscle actin+/Col-1+ peribronchial myofibroblasts by immunohistochemistry. Corticosteroids significantly reduced allergen-induced increases in peribronchial collagen deposition and levels of total lung collagen but did not reduce allergen-induced increases in the thickness of the peribronchial smooth muscle layer. Levels of lung TGF-beta1 were significantly reduced in mice treated with systemic corticosteroids, and this was associated with a significant decrease in the number of peribronchial inflammatory cells that expressed TGF-beta1, including eosinophils and mononuclear cells. Corticosteroids also significantly reduced the number of peribronchial myofibroblasts. Overall, these studies demonstrate that administration of corticosteroids significantly reduces levels of allergen-induced peribronchial fibrosis. The reduction in peribronchial fibrosis mediated by corticosteroids is likely to be due to several mechanisms including inhibition of expression of TGF beta1, a reduction in the number of peribronchial inflammatory cells expressing TGF-beta1 (eosinophils, macrophages), as well as by corticosteroids reducing the accumulation of peribronchial myofibroblasts that contribute to collagen expression. PMID- 16344334 TI - Interstitial lung disease: effects of thin-section CT on clinical decision making. AB - PURPOSE: To retrospectively quantify the change in the diagnosis and management of suspected interstitial lung disease when thin-section computed tomography (CT) is added to pretest probabilities. MATERIALS AND METHODS: The institutional review board does not require approval or patient informed consent for retrospective study of case records and CT studies. Six pulmonologists reviewed data sheets containing clinical information and results of pulmonary function tests and chest radiographs of 168 consecutive patients (86 women and 82 men; mean age, 59.8 years; age range, 22-86 years) suspected of having interstitial lung disease. Differential diagnoses and responses to specific questions regarding patient care were recorded before and after assimilation of thin section CT findings. Both unweighted and weighted kappa analyses were used to determine agreement between pulmonologists before and after CT. RESULTS: First choice diagnosis changed in 520 (51%) of 1008 cases, and agreement on first choice diagnosis increased from 0.47 to 0.72 after thin-section CT. In addition, confidence in the first-choice diagnosis increased, and there was a reduction in the number of differential diagnoses offered by all pulmonologists (P < .005 and P < .001, respectively). Agreement on diagnostic probabilities for individual disorders increased substantially, particularly for diagnoses of idiopathic pulmonary fibrosis (weighted kappa = 0.58-0.89). With CT findings, pulmonologists changed their pre-CT responses regarding the use of bronchoalveolar lavage, transbronchial biopsy, and thoracoscopic biopsy in 242 (24.0%), 282 (28.0%), and 292 (29.0%) of 1008 cases, respectively. However, agreement for the use of these investigations was low both before and after CT. The request rate for thoracoscopic biopsy in patients in whom idiopathic fibrosis was diagnosed decreased from 48 of 179 (26.8%) to 26 of 233 (11.2%) after CT. CONCLUSION: Thin section CT resulted in a change in first-choice diagnosis in half the cases. Diagnostic confidence improved, and CT findings increased agreement between pulmonologists on diagnostic probabilities across a range of interstitial lung diseases. PMID- 16344335 TI - Prediction of organ-confined prostate cancer: incremental value of MR imaging and MR spectroscopic imaging to staging nomograms. AB - PURPOSE: To assess retrospectively the incremental value of endorectal coil magnetic resonance (MR) imaging and combined endorectal MR imaging-MR spectroscopic imaging to the staging nomograms for predicting organ-confined prostate cancer (OCPC). MATERIALS AND METHODS: The institutional review board approved this HIPAA-compliant study and issued a waiver of informed consent for review of the MR reports and clinical data. Between November 1, 1999, and November 1, 2004, 229 patients underwent endorectal MR imaging and 383 underwent combined endorectal MR imaging-MR spectroscopic imaging before radical prostatectomy. Mean patient age was 58 years (range, 32-74 years). MR studies were interpreted prospectively by 12 radiologists who were informed of patients' clinical data. On the basis of the MR reports, the risks of extracapsular extension, seminal vesicle invasion, and lymph node metastasis were scored retrospectively from 1 to 5; the highest score was subtracted from 6 to determine a score (from 1 to 5) for the likelihood of OCPC on MR studies. The staging nomograms were used to calculate the likelihood of OCPC on the basis of serum prostate-specific antigen level, Gleason grade at biopsy, and clinical stage. Histopathologic findings constituted the reference standard. Logistic regression was used to estimate the multivariable relations between OCPC and MR findings. The area under the receiver operator characteristic curve was calculated for each model. The jackknife method was used for bias correction. RESULTS: MR findings contributed significant incremental value (P A nucleotide change was a common mutation, present in 35% of the mutated chromosomes. Clinical presentations included asymptomatic hyperCKemia, severe early-onset muscular dystrophy, and mild late-onset muscular dystrophy. Dilated cardiomyopathy and ventilatory impairment were frequent features. Significant intrafamilial and interfamilial clinical variability was observed. CONCLUSIONS: FKRP mutations are a frequent cause of limb-girdle muscular dystrophies. The degree of respiratory and cardiac insufficiency in patients did not correlate with the severity of muscle involvement. The finding of 2 asymptomatic patients with FKRP mutations suggests that modulating factors may ameliorate the clinical phenotype. PMID- 16344348 TI - Brain morphometry, T2-weighted hyperintensities, and IQ in children with neurofibromatosis type 1. AB - BACKGROUND: Larger gray matter (GM) volume in healthy children is correlated with higher IQ. Children with neurofibromatosis type 1 (NF1) have larger brains, their magnetic resonance images frequently show T2-weighted hyperintensities, and their IQs are lower. OBJECTIVES: To confirm the hypotheses that (1) children with NF1 have larger GM and white matter volumes, (2) the greatest volume differences are in the frontal and parietal regions and in children with NF1 with hyperintensities, and (3) GM volume is inversely related to IQ in children with NF1. DESIGN: Wechsler Intelligence Scale for Children-Third Edition IQ testing and measurement of cerebral volumes and hyperintensities in brain magnetic resonance images were performed on 36 children with NF1 and on 36 matched relatives who served as control subjects. RESULTS: Gray matter and white matter volumes were significantly larger in children with NF1. The greatest difference was observed in cerebral white matter volume, predominantly in the frontal lobes, whereas the greatest difference in GM volume was in the temporal, parietal, and occipital regions. In controls, IQ was significantly related to GM volume, but in children with NF1, IQ was not inversely associated with GM volume, although IQs of children with NF1 were significantly lower. CONCLUSIONS: Children with NF1 do not have the normal relationship between GM volume and IQ. Larger GM volume in the posterior brain regions and larger white matter volumes in the frontal brain regions contribute to the larger brain volume in children with NF1. PMID- 16344349 TI - Central nervous system involvement in hereditary neuropathy with liability to pressure palsies: description of a large family with this association. AB - OBJECTIVE: To describe a large family with hereditary neuropathy with liability to pressure palsies associated with central nervous system demyelination. DESIGN: We examined the 18 members of a pedigree. Genetic analysis was performed on 15 subjects, standard nerve conduction studies on 10 subjects, and brain magnetic resonance imaging studies on 8 subjects. RESULTS: Hereditary neuropathy with liability to pressure palsies was confirmed in 9 patients of the pedigree. Brain magnetic resonance imaging findings showed multiple areas of demyelination in 6 of 6 affected members and were normal in 2 of 2 healthy relatives. Magnetic resonance imaging abnormalities were predominantly located in the subcortical frontal white matter. All patients had acute and recurrent nerve palsies, while clinical features of central nervous system involvement were not a characteristic of this pedigree. CONCLUSIONS: We demonstrate that this association, previously reported in sporadic cases, is not coincidental. Therefore, patients with hereditary neuropathy with liability to pressure palsies can present central nervous system white matter lesions, and the role of the PMP22 (peripheral myelin protein 22) gene deletion in the central nervous system should be further studied. PMID- 16344350 TI - Subclavian artery dissection and triple infarction of the nervous system. AB - BACKGROUND: Subclavian artery dissection is a rare entity. It is usually associated with anomalous aortic vasculature. Only with trauma or catheterization procedures is subclavian artery dissection with normal aortic vasculature reported. PATIENT: We describe a patient with intrascapular pain, an occipital headache, and 3 distinct infarctions in the nervous system. He had spontaneous subclavian artery dissection with normal aortic vasculature. CONCLUSION: Subclavian artery dissections should be suspected in patients with intrascapular pain, occipital or cervical pain, and symptoms within the posterior circulation. PMID- 16344351 TI - Clinical and molecular features of encephalomyopathy due to the A3302G mutation in the mitochondrial tRNA(Leu(UUR)) gene. AB - BACKGROUND: The mitochondrial DNA mutation A3302G in the tRNA(Leu(UUR)) gene causes respiratory chain complex I deficiency. The main clinical feature appears to be a progressive mitochondrial myopathy with proximal muscle weakness. OBJECTIVE: To report on clinical and molecular features in 4 novel patients with the A3302G mutation. DESIGN: Case reports. PATIENTS: Four patients (3 of whom are from the same family) with a myopathy caused by the A3302G mitochondrial DNA mutation. MAIN OUTCOME MEASURE: Identification of the A3302G mutation by DNA sequencing. RESULTS: All 4 patients had an adult-onset progressive mitochondrial myopathy with proximal muscle weakness, resulting in exercise intolerance. In 2 unrelated patients, upper limb reflexes were absent with preservation of at least some lower limb reflexes. Other features including hearing loss, recurrent headaches, ptosis, progressive external ophthalmoplegia, and depression were present. CONCLUSION: While the dominant clinical features of the A3302G mutation were exercise intolerance and proximal muscle weakness, other features of mitochondrial encephalomyopathies, previously not described for this mutation, were present. PMID- 16344352 TI - Sturge-Weber syndrome associated with other abnormalities: a medical record and literature review. AB - OBJECTIVE: To develop hypotheses regarding the relationship between Sturge-Weber syndrome (SWS) and other abnormalities in a subset of patients. DESIGN: We retrospectively reviewed medical records in a group of 28 patients with SWS, noting the main features of SWS and accompanying unexpected abnormalities. We also conducted a literature review of abnormalities associated with SWS. RESULTS: Twenty-eight medical records of patients with SWS were reviewed. Of this number, we found 8 (29%, 2 female) patients who manifested other abnormalities. Our review of the literature uncovered 15 additional cases with associated abnormalities. CONCLUSIONS: We hypothesize that the abnormalities associated with SWS suggest testable insights regarding pathogenesis and that chromosome 17p1-p13 may be a candidate region for genes involved with SWS. We also propose that some patients with SWS may have disorders of cholesterol biosynthesis or carbohydrate glycosylation. PMID- 16344353 TI - Sturge-Weber syndrome and accompanying Dyke-Davidoff-Masson syndrome. PMID- 16344354 TI - Relapsing focal myositis: the localization detected by gallium citrate Ga 67 scintigraphy. PMID- 16344355 TI - Uncal herniation. PMID- 16344356 TI - What is deep brain stimulation "failure" and how do we manage our own failures? PMID- 16344357 TI - Bread-and-butter in diagnosis of myasthenia gravis. PMID- 16344358 TI - Otitic hydrocephalus. PMID- 16344360 TI - Low muscular mass and overestimation of microalbuminuria by urinary albumin/creatinine ratio. AB - Microalbuminuria is a mild urinary albumin elevation and is associated with cardiovascular disease. Urinary albumin/creatinine ratio is recommended for microalbuminuria assessment, because it reflects urinary albumin excretion. Muscular mass could affect albumin/creatinine ratio, because urinary creatinine reflects muscular mass. The study investigated high albumin/creatinine ratio attributed to low urinary creatinine without microalbuminuria. The Gubbio Population Study for ages 45 to 64 collected data on weight, skinfold, urinary albumin, urinary creatinine, and coronary heart disease. Weight and skinfold thickness were used to calculate fat and nonfat mass and urinary creatinine as a marker of muscular mass. Microalbuminuria was defined as urinary albumin of 20 to 199 microg/min and high albumin/creatinine ratio as a ratio of 17 to 250 microg/mg in men and of 25 to 355 microg/mg in women. Persons with macroalbuminuria (urinary albumin > or =200 microg/min) were excluded to focus analyses on microalbuminuria. Coronary heart disease was defined by ECG and questionnaire. The target cohort consisted of 1623 men and women, ages 45 to 64. Prevalence was 8.5% for high albumin/creatinine ratio (n=138), 4.3% for microalbuminuria (n=69), 5.2% for high albumin/creatinine ratio without microalbuminuria (n=85), and 1.0% for nonhigh albumin/creatinine ratio with microalbuminuria (n=16). High albumin/creatinine ratio without microalbuminuria was inversely associated with nonfat mass and urinary creatinine (P<0.04). Compared with persons with a nonhigh albumin/creatinine ratio, coronary heart disease was more prevalent in persons with a high albumin/creatinine ratio and microalbuminuria (18.9% and 7.1%; P=0.002), not in persons with a high albumin/creatinine ratio without microalbuminuria (8.2% and 7.1%; P=0.706). A high albumin/creatinine ratio in persons with low muscle mass indicates low urinary creatinine more often than microalbuminuria and cardiovascular disease. PMID- 16344361 TI - System y+ arginine transport and NO production in peripheral blood mononuclear cells in pregnancy and preeclampsia. AB - Systemic inflammation and oxidative stress are features of normal pregnancy and, in excess, contribute to the pathogenesis of preeclampsia. Inflammatory cell activation stimulates uptake of arginine (the precursor for nitric oxide) by transport system y+, expression of one of its genes (CAT-2) together with inducible nitric oxide synthase, leading to nitric oxide production. We investigated whether these changes occur in peripheral blood mononuclear cells in normal pregnancy and are exaggerated in preeclampsia. Samples from matched trios of nonpregnant, normal pregnant, and preeclamptic women were studied. Arginine transport was characterized, and the expression of inducible nitric oxide synthase and cell-specific nitric oxide production were measured. Arginine uptake by system y+ was significantly increased (P<0.001) in peripheral blood mononuclear cells in normal pregnancy but not in preeclampsia. CAT-2 mRNA was not detected in cells from nonpregnant women but was detected in 3 of 10 normal pregnant and 8 of 10 of preeclamptic women (P<0.001). Inducible nitric oxide synthase protein expression was significantly increased in normal pregnant women (P<0.05) but not preeclamptic women. No significant differences in cell-specific nitric oxide production were observed. These changes confirm the predictions for normal pregnancy but not for preeclampsia in which, despite increases in CAT-2 expression, arginine uptake is not additionally increased. This may create a relative deficiency of arginine in PBMCs favoring superoxide and peroxynitrite production and contribute to oxidative and nitrosative stress in preeclampsia. PMID- 16344362 TI - Novel integrative approaches to the identification of candidate genes in hypertension. AB - Hypertension, diabetes, and obesity are common diseases that are genetically expressed as complex traits. The clinical expression of these diseases, which run in families but have no clear pattern of inheritance, has been attributed to the interaction between multiple genes and the environment. Identifying the underlying genes, a crucial step in understanding the molecular pathogenesis of complex diseases, has had limited success so far, stressing the need for novel strategies to move this process forward. Innovative integrative genetic-genomic approaches have been proposed recently for the identification of new high priority candidate genes. These strategies, which combine expression profiling with genetic linkage in different ways, may represent a breakthrough in the search for the genes involved in complex diseases. PMID- 16344363 TI - Sympathetic neural outflow and chemoreflex sensitivity are related to spontaneous breathing rate in normal men. AB - Respiration contributes importantly to short-term modulation of sympathetic nerve activity. However, the relationship between spontaneous breathing rate, chemoreflex function, and direct measures of sympathetic traffic in healthy humans has not been studied previously. We tested the hypothesis that muscle sympathetic nerve activity and chemoreflex sensitivity are linked independently to respiratory rate in normal subjects. We studied 69 normal male subjects aged 29.6+/-8.1 years. Subjects were subdivided according to the tertiles of respiratory rate distributions. Mean respiration rate was 10.6 breaths/min in the first tertile, 14.8 breaths/min in the second tertile, and 18.0 breaths/min in the third tertile. Subjects from the third tertile (faster respiratory rate) had greater sympathetic activity than subjects from the first tertile (slower respiratory rate; 29+/-3 versus 17+/-2 bursts/min; P<0.001). Stepwise multiple linear regression analysis revealed that only respiratory rate was linked independently to sympathetic activity (r=0.42; P<0.001). In comparison to subjects with slow respiratory rate, subjects with fast respiratory rate had greater increases in minute ventilation during both hypercapnia (7.3+/-0.8 versus 3.2+/-1.0 L/min; P=0.005) and hypoxia (5.7+/-0.8 versus 2.4+/-0.7 L/min; P=0.007). Muscle sympathetic nerve activity and chemoreflex sensitivity are linked to spontaneous respiratory rate in normal humans. Faster respiratory rate is associated with higher levels of sympathetic traffic and potentiated responses to hypoxia and hypercapnia. Spontaneous breathing frequency, central sympathetic outflow, and chemoreflex sensitivity exhibit significant and hitherto unrecognized interactions in the modulation of neural circulatory control. PMID- 16344364 TI - Cost-effectiveness of ambulatory blood pressure: a reanalysis. AB - Accurate diagnosis of hypertension and prognosis for future cardiovascular events can be enhanced through the use of 24-hour ambulatory blood pressure monitoring. It has been suggested that the use of ambulatory monitoring as a secondary screening for hypertension might be cost-effective. Many needed studies that are related to the calculation of cost-effectiveness for ambulatory monitoring have become available in recent years. More accurate estimates for cost of care, costs for testing, prevalence of white-coat hypertension, and incidence of the transition from normal pressures to hypertension have been reported. This study presents calculations of the cost savings likely to take place when ambulatory blood pressure monitoring is implemented for newly detected hypertensive subjects. These calculations are based on current estimates for cost of testing, cost of treatment, prevalence of white-coat hypertension at baseline, and varying the incidence of new hypertension after the initial screening. The results indicate a potential savings of 3% to 14% for cost of care for hypertension and 10% to 23% reduction in treatment days when ambulatory blood pressure monitoring is incorporated into the diagnostic process. At current reimbursement rates, the cost of ambulatory blood pressure monitoring for secondary screening on an annual basis would be <10% of treatment costs. Calculated savings for use of ambulatory blood pressure monitoring can take place when annual treatment costs are as little as 300 dollars. These estimates should be considered for the management of recently detected hypertension, especially when the risk of future cardiovascular is disease is low. PMID- 16344365 TI - Expanding the use of ambulatory blood pressure monitoring for the diagnosis and management of patients with hypertension. PMID- 16344366 TI - Reduced NAD(P)H oxidase in low renin hypertension: link among angiotensin II, atherogenesis, and blood pressure. AB - Endothelial dysfunction (ED) complicates hypertension and is a precursor of atherosclerosis. Reduced NO bioactivity, because of increased reduced NAD(P)H oxidase-derived reactive oxygen species (ROS), plays a critical role in ED. gp91phox, predominantly expressed in the endothelium and adventitia, is a subunit of NAD(P)H oxidase important for its activation in response to angiotensin (Ang) II. Human atherosclerotic plaques are heavy laden with gp91phox. We have shown that in Dahl salt-sensitive (DS) rats, a paradigm of low renin salt-sensitive (SS) hypertension in humans, Ang II receptor blockade normalizes ROS production and endothelium-dependent relaxation (EDR) without significantly affecting systolic blood pressure (SBP). To additionally elucidate the mechanisms involved in the functional association of Ang II in SS hypertension, we administered a cell-permeable inhibitor of the assembly of p47phox with gp91phox in NAD(P)H oxidase, gp91ds-tat (10 mg/kg body weight, 3 weeks by minipump), to DS rats fed a 4% salt diet. Control rats received either vehicle or an inactive scramb-tat peptide. Vehicle-treated DS developed hypertension (SBP 168+/-5 mm Hg), left ventricular hypertrophy (LVH), proteinuria, impaired EDR, and increased aortic ROS production (superoxide 115% and peroxynitrite 157%) and expression of the proatherogenic molecules LOX-1 (130%) and MCP-1 (166%). gp91ds-tat, but not scramb-tat, normalized ROS and EDR, as well as LOX-1 and MCP-1, despite nonsignificant effects on SBP (159+/-5 mm Hg; P>0.05), left ventricular hypertrophy, and proteinuria. Our findings support the notion that in SS hypertension, activation of NAD(P)H oxidase promotes ED and atherogenesis via decreased nitric oxide bioactivity and increased LOX-1 and MCP-1, independent of blood pressure. PMID- 16344367 TI - Tetrahydrobiopterin, but not L-arginine, decreases NO synthase uncoupling in cells expressing high levels of endothelial NO synthase. AB - Endothelial NO synthase (eNOS) produces superoxide when depleted of (6R)-5,6,7,8 tetrahydro-L-biopterin (BH4) and L-arginine by uncoupling the electron flow from NO production. High expression of eNOS has been reported to have beneficial effects in atherosclerotic arteries after relatively short periods of time. However, sustained high expression of eNOS may have disadvantageous vascular effects because of uncoupling. We investigated NO and reactive oxygen species (ROS) production in a microvascular endothelial cell line (bEnd.3) with sustained high eNOS expression and absent inducible NOS and neuronal NOS expression using 4,5-diaminofluorescein diacetate and diacetyldichlorofluorescein as probes, respectively. Unstimulated cells produced both NO and ROS. After stimulation with vascular endothelial growth factor (VEGF), NO and ROS production increased. VEGF induced ROS production was even further increased by the addition of extra L arginine. Nomega-nitro-L-arginine methyl ester decreased ROS production. These findings strongly suggest that eNOS is a source of ROS in these cells. Although BH4 levels were increased as compared with another endothelial cell line, eNOS levels were >2 orders of magnitude higher. The addition of BH4 resulted in increased NO production and decreased generation of ROS, indicating that bEnd.3 cells produce ROS through eNOS uncoupling because of relative BH4 deficiency. Nevertheless, eNOS-dependent ROS production was not completely abolished by the addition of BH4, suggesting intrinsic superoxide production by eNOS. This study indicates that potentially beneficial sustained increases in eNOS expression and activity could lead to eNOS uncoupling and superoxide production as a consequence. Therefore, sustained increases of eNOS or VEGF activity should be accompanied by concomitant supplementation of BH4. PMID- 16344368 TI - A genetic locus accentuates the effect of volume overload on adverse left ventricular remodeling in male and female rats. AB - Although increased left ventricular (LV) mass is highly predictive of cardiovascular morbidity and mortality in humans, it has never been verified in an experimental model that naturally occurring alleles linked to increased LV mass under basal conditions also associate with worsened cardiovascular prognosis. Because we have shown previously that locus Cm24 on chromosome 5 was responsible for differences in LV mass between WKY and WKHA rats, we used WKY.WKHA-(D5Rat45-D5Rat245) congenic rats (where locus Cm24 has been transferred from WKHA into WKY rats) to test how naturally occurring gene variants present in Cm24 would, in addition to their effects under basal conditions, affect LV mass remodeling and/or function in the context of overload. Volume overload was induced in WKY, WKHA, and WKY.WKHA congenic rats by surgical creation of an aorto caval fistula. In females, the fistula had no effect on the hearts of WKY rats, yet it induced dilated eccentric hypertrophy and isolated diastolic dysfunction in WKHA and WKY.WKHA congenic rats, along with signs of congestive heart failure. In males, the surgical maneuver induced only mild or inconsistent responses in WKY rats but had much more pronounced effects in WKHA and WKY.WKHA congenic rats. Altogether, our data show that a genetic locus that induces, under basal conditions, either mild or no concentric LV remodeling in either male or female rats, respectively, associates with LV dilatation and dysfunction in both sexes when the hearts are additionally challenged. PMID- 16344369 TI - Phagocytosis of necrotic but not apoptotic trophoblasts induces endothelial cell activation. AB - It is hypothesized that preeclampsia is caused by factors from the placenta that induce endothelial cell activation. Trophoblasts are cells that may be shed from the placenta, then deported in the maternal blood, and finally become trapped in the pulmonary capillaries. The ultimate fate of deported trophoblasts is unknown, but to prevent clogging of the pulmonary circulation they must be cleared from the capillary beds. We examined the hypothesis that endothelial cells phagocytose deported trophoblasts and also examined the consequent effects of the trophoblasts on endothelial cells. Fluorescently labeled trophoblast-derived choriocarcinoma cells were induced to become apoptotic or necrotic and exposed to endothelial cell monolayers. Confocal microscopy demonstrated uptake of both apoptotic and necrotic trophoblasts, and this phagocytosis could be inhibited by cytochalasin B. Phagocytosis of necrotic but not apoptotic trophoblasts induced increased endothelial intercellular adhesion molecule 1 (ICAM-1) expression, as well as increased adhesion of monocytes to endothelial cell monolayers. Inhibiting the phosphatidylinositol 3-kinase and p38 mitogen-activated protein kinase pathways blocked both expression of ICAM-1 and phagocytosis, whereas inhibition of the P42/44 mitogen-activated protein kinase pathway blocked only ICAM-1 expression. This work suggests that endothelial cells can phagocytose deported trophoblasts and that the mechanism of trophoblast death (apoptotic or necrotic) could have major effects on the maternal vascular response to shed trophoblasts. PMID- 16344370 TI - Lipid rafts take center stage in endothelial cell redox signaling by death receptors. PMID- 16344371 TI - Peroxisome proliferator-activated receptor gamma regulates angiotensin II stimulated phosphatidylinositol 3-kinase and mitogen-activated protein kinase in blood vessels in vivo. AB - Angiotensin (Ang) II is implicated in hypertension, vascular remodeling, and insulin resistance. Peroxisome proliferator-activated receptor (PPAR) gamma activators increase insulin sensitivity and improve Ang II-induced vascular remodeling. We evaluated the effects of the PPAR-gamma activator rosiglitazone on Ang II signaling in aorta and mesenteric arteries. Rats received Ang II by subcutaneous infusion and/or rosiglitazone per os for 7 days. Blood pressure rise in Ang II-infused rats was attenuated by rosiglitazone. Ang II significantly increased Ang II type 1 receptor expression in the mesenteric arteries (P<0.001), whereas that of the aorta was decreased (P<0.05), changes which were reversed by rosiglitazone. Akt activity was increased by Ang II and returned to basal levels under rosiglitazone in both vascular beds. However, Ang II-induced extracellular signal-regulated kinase 1/2 activity increased in aorta but not in mesenteric vessels (P<0.001), where 4E-binding protein 1 activity was significantly increased by Ang II and inhibited by PPAR-gamma activation. In response to Ang II, Src homology (SH) 2-containing inositol phosphatase 2 activity was increased (P<0.05) in both vascular beds. In conclusion, PPAR-gamma activator rosiglitazone attenuated Ang II-induced blood pressure elevation and intracellular signaling on aorta and mesenteric vessels. There was differential inhibition of Ang II type 1 receptor receptors/phosphatidylinositol 3-kinase/Akt and extracellular signal regulated kinase 1/2 in both vessels. Effects of PPAR-gamma activators on these pathways could contribute to regression of vascular remodeling in models of hypertension and diabetes and, accordingly, in hypertensive diabetic patients. PMID- 16344372 TI - Lipid raft clustering and redox signaling platform formation in coronary arterial endothelial cells. AB - Recent studies have indicated that lipid rafts (LRs) in the cell membrane are clustered in response to different stimuli to form signaling platforms for transmembrane transduction. It remains unknown whether this LR clustering participates in redox signaling in endothelial cells. The present study tested a hypothesis that clustering of LRs on the membrane of coronary endothelial cells produces aggregation and activation of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase, thereby forming a redox signaling platform. By confocal microscopic analysis of agonist-stimulated rafts patch formation, we found that several death receptor ligands or apoptotic factors, including tumor necrosis factor alpha, Fas ligand, or endostatin, stimulated the clustering and trafficking of individual LRs on the plasma membrane of coronary endothelial cells. Interestingly, double labeling of a membrane-bound NADPH oxidase subunit, gp91phox, and LRs showed that gp91phox colocalized within the LR patches when endothelial cells were stimulated by Fas ligand. In isolated LR fractions from Fas-stimulated endothelial cells, gp91phox, p47phox (a crucial cytosolic regulatory subunit of NADPH oxidase), and Rac GTPase were markedly increased and blocked by nystatin, a compound that disrupts LRs. These clustered LRs contained high NADPH oxidase activity, which increased in response to Fas stimulation. Functionally, Fas ligand-induced inhibition of endothelium-dependent vasorelaxation was reduced if LRs were disrupted or NADPH oxidase was inhibited. These results suggest that LR clustering occurs in coronary endothelial cells. The formation of redox signaling platforms on the cell membrane mediates transmembrane signaling of death receptors, resulting in endothelial dysfunction. PMID- 16344373 TI - Automated oscillometric determination of the ankle-brachial index provides accuracy necessary for office practice. AB - Peripheral arterial disease (PAD) remains underdiagnosed by primary care and cardiovascular physicians. The office-based assessment of PAD is limited by the need for specialized equipment and the time required for performance of the ankle brachial index (ABI). We explored whether the accuracy of automated ABI measurement by oscillometry compared favorably with the gold-standard method using continuous-wave Doppler ultrasound. Consecutive patients referred to our university hospital noninvasive vascular laboratory for ABI measurement were invited for participation. Of 205 patients, 201 participated, including 55 with PAD. The ABI was measured by automated oscillometry and Doppler ultrasound. The test of trends revealed a correlation coefficient of 0.78 in the left leg and 0.78 in the right leg (P<0.01 for both). The mean ABI difference between methods was 0.04+/-0.01 and 0.06+/-0.01, respectively, in the left and right legs. The differences between the methods followed a normal distribution. Oscillometric determination of the ABI provides an accurate determination of the ABI in an outpatient population. Our findings show automated oscillometry to be a reliable and easier method of ABI measurement, lowering the barrier to incorporation of this diagnostic test into clinical practice. PMID- 16344374 TI - Potential of gene therapy strategy for the treatment of hypertension. PMID- 16344375 TI - Dyslipidemia and the risk of incident hypertension in men. AB - Evidence suggests that hypertension may share a similar pathophysiology with cardiovascular disease (CVD). Thus, dyslipidemia, a strong predictor of CVD, may also predict incident hypertension. We analyzed 3110 men free of hypertension, CVD, and cancer from the Physicians' Health Study, who provided baseline blood samples from which we measured total cholesterol (TC) and HDL cholesterol (HDL C), and calculated non-HDL-C and the TC/HDL-C ratio. We categorized each lipid parameter into quintiles and considered National Cholesterol Education Project clinical cut points. Other risk factor information was provided from self-reports on the baseline questionnaire. Incident hypertension was defined as either the initiation of antihypertensive treatment, self-reported systolic blood pressure > or =140 mm Hg, or diastolic blood pressure > or =90 mm Hg. Over a mean follow-up of 14.1 years, 1019 men developed hypertension. In Cox proportional hazards models adjusted for lifestyle and clinical risk factors, men in the highest quintile of TC, non-HDL-C, and TC/HDL-C ratio had increased risks of developing hypertension of 23%, 39%, and 54%, respectively, compared with participants in the lowest quintile. Furthermore, men in the highest quintile of HDL-C had a 32% decreased risk of developing hypertension compared with those in the lowest quintile. Models using National Cholesterol Education Project cut points demonstrated similar associations with hypertension. Models excluding men with diabetes and obesity maintained an independent association between baseline lipids and hypertension. These prospective cohort data suggest that dyslipidemias may lead to the subsequent development of hypertension. Thus, plasma lipids may be useful in the identification of men at risk for hypertension. PMID- 16344376 TI - Out-of-hemodialysis-unit blood pressure is a superior determinant of left ventricular hypertrophy. AB - Blood pressures (BPs) obtained in the dialysis unit correlate poorly with ambulatory BP and left-ventricular hypertrophy (LVH). We compared the performance of BP obtained within and outside the dialysis unit as a correlate of LVH. BP was obtained in the dialysis unit using routine and standardized methods and outside the dialysis unit using home and ambulatory BP monitoring in 140 patients (mean age, 56 years; 89 men; 129 blacks; and 59 with diabetes mellitus) on chronic hemodialysis for > or =3 months. Dialysis unit BP recordings were averaged over 2 weeks, and home BP averaged over 1 week. Ambulatory BP monitoring was performed during an interdialytic interval. Echocardiography was performed immediately after dialysis for the assessment of left-ventricular mass. Left ventricular mass/height(2.7) of >51 g/m2 was taken as evidence of LVH. Test performance of various BPs was compared using receiver operating characteristic curves. Average ambulatory BP was 129.7+/-21.2/73.6+/-13.1 mm Hg, home BP was 139.4+/-21.2/79.0+/ 12.5 mm Hg, standardized predialysis BP was 142.1+/-21.7/74.9+/-13.3 mm Hg, postdialysis was 120.9+/-20.8/69.6+/-12.5 mm Hg, routine predialysis was 145.6+/ 20.7/79.4+/-13.1 mm Hg, and postdialysis was 132.0+/-19.3/72.6+/-11.1 mm Hg. Left ventricular mass/height(2.7) was 59.1+/-16.5, and 68% had LV hypertrophy. Diastolic BP measured by any technique was not associated with LVH. Routine and standardized measurements of BP were similarly weak correlates of LVH. Systolic BP outside the dialysis unit was a stronger correlate of LVH compared with dialysis unit BP. PMID- 16344377 TI - NOS3 regulation: renal tubular epithelial cells are not simply large endothelial cells. PMID- 16344378 TI - A high-salt diet dissociates NO synthase-3 expression and NO production by the thick ascending limb. AB - NO produced by endothelial NO synthase (NOS3) decreases sodium transport by the thick ascending limb (THAL). We found previously that 7 days of high salt (HS) increased THAL-NOS3 expression but not NO production. NOS3 phosphorylation regulates enzyme activity. We hypothesized that HS acutely increases NOS3 expression and NO production, and, over time, changes in NOS3 phosphorylation dissociate NO production from expression. NOS3 expression increased by 71+/-13%, 127+/-24%, and 69+/-16% at days 1, 3, and 7 of HS, respectively. At days 14 and 28, expression was back to normal salt. After 1 day of HS, NO production in response to 250 micromol/L L-arginine was elevated by 146% and, by day 3, returned to normal salt. Similar increases were found in response to endothelin 1. Inhibitors of NOS1/2 did not blunt the salt-induced increase in NO. Phosphorylation at Thr495, an inhibitory site, decreased by 39+/-8% at day 1 of HS and then increased by 116+/-18% at day 3. Phosphorylation at Ser633 and Ser1177 (stimulatory sites) decreased by &25% at day 1 and remained depressed at day 3. Superoxide production increased by 71% at day 1, decreased by 57% at day 3, and decreased by 55% at day 7. The NOS inhibitor L-NG-nitroarginine methyl ester did not alter superoxide levels at any time point. The addition of reduced nicotinamide-adenine dinucleotide phosphate and tetrahydrobiopterin had no effect on NO release after 3 days of HS. We conclude the following: (1) HS transiently increases NO production and NOS3 expression; (2) NOS3 expression and NO production are dissociated by HS; and (3) changes in phosphorylation explain how THAL NOS3 activity and expression are dissociated by HS. PMID- 16344379 TI - Treatment of hypertension: remaining issues after the Anglo-Scandinavian Cardiac Outcomes Trial. PMID- 16344380 TI - The impact of the Canadian Hypertension Education Program on antihypertensive prescribing trends. AB - Although previous studies have shown that hypertension management has improved in Canada during the past decade, this study was designed to determine whether these changes were temporally related to initiation of the Canadian Hypertension Education Program in 1999. Antihypertensive prescription rates in Ontario were compared using time series analyses before and after 1999 in 2 Ontario cohorts: all hypertensives prescribed therapy (using the Intercontinental Medical Statistics CompuScript Database) and all elderly hypertensives without diabetes prescribed therapy (using linked administrative databases including the Ontario Drug Benefit Database). Between January 1998 and December 2003, &280 million prescriptions for antihypertensive agents were filled in Ontario, and total antihypertensive prescriptions increased by 58% annually; time series analyses confirmed that the prescribing rates for total antihypertensives, thiazide diuretics, beta-blockers, and calcium channel blockers increased significantly after 1999, even after adjustment for the temporal trends in the pre-1999 data. In the 166,018 nondiabetic individuals over age 65 who were newly treated for hypertension in Ontario between July 1994 and March 2002, changes in prescription rates for total antihypertensive drugs, angiotensin-converting enzyme inhibitors, beta blockers, and calcium channel blockers occurred in directions that were consistent with guideline recommendations and were statistically significantly related to the initiation of the Canadian Hypertension Education Program. The substantial changes in prescription rates for guideline-recommended antihypertensive drug classes in elderly Ontarians without diabetes and the general Ontario population seen in the past decade are temporally related to the initiation of the Canadian Hypertension Education Program. PMID- 16344381 TI - Unfractionated and low-molecular-weight heparin as adjuncts to thrombolysis in aspirin-treated patients with ST-elevation acute myocardial infarction: a meta analysis of the randomized trials. AB - BACKGROUND: There is uncertainty about the role of intravenous unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) in patients with ST elevation myocardial infarction (STEMI) treated with aspirin and thrombolysis. METHODS AND RESULTS: We performed a meta-analysis of the randomized trials to assess the effect of UFH and LMWH on reinfarction, death, stroke, and bleeding. Fourteen trials involving a total of 25,280 patients were included (1239 comparing intravenous UFH versus placebo or no heparin; 16,943 comparing LMWH versus placebo; and 7098 comparing LMWH versus intravenous UFH). Intravenous UFH during hospitalization did not reduce reinfarction (3.5% versus 3.3%; odds ratio [OR], 1.08; 95% CI, 0.58 to 1.99) or death (4.8% versus 4.6%; OR, 1.04; 95% CI, 0.62 to 1.78) and did not increase major bleeding (4.2% versus 3.4%; OR, 1.21; 95% CI, 0.67 to 2.18) but increased minor bleeding (19.6% versus 12.5%; OR, 1.72; 95% CI, 1.22 to 2.43). During hospitalization/at 7 days, LMWH compared with placebo reduced the risk of reinfarction by approximately one quarter (1.6% versus 2.2%; OR, 0.72; 95% CI, 0.58 to 0.90; number needed to treat [NNT]=167) and death by &10% (7.8% versus 8.7%; OR, 0.90; 95% CI, 0.80 to 0.99; NNT=111) but increased major bleeding (1.1% versus 0.4%; OR, 2.70; 95% CI, 1.83 to 3.99; number needed to harm [NNH]=143) and intracranial bleeding (0.3% versus 0.1%; OR, 2.18; 95% CI, 1.07 to 4.52; NNH=500). The reduction in death with LMWH remained evident at 30 days. LMWH compared with UFH during hospitalization/at 7 days reduced reinfarction by &45% (3.0% versus 5.2%; OR, 0.57; 95% CI, 0.45 to 0.73; NNT=45), did not reduce death (4.8% versus 5.3%; OR, 0.92; 95% CI, 0.74 to 1.13) or increase major bleeding (3.3% versus 2.5%; OR, 1.30; 95% CI, 0.98 to 1.72), but increased minor bleeding (22.8% vs 19.4%; OR, 1.26; 95% CI, 1.12 to 1.43). The reduction in reinfarction remained evident at 30 days. CONCLUSIONS: In aspirin-treated patients with STEMI who are treated with thrombolysis, intravenous UFH has not been shown to prevent reinfarction or death. LMWH given for 4 to 8 days compared with placebo reduces reinfarction by approximately one quarter and death by &10% and when directly compared with UFH reduces reinfarction by almost one half. These data suggest that LMWH should be the preferred antithrombin in this setting. PMID- 16344382 TI - Pharmacological preconditioning with tumor necrosis factor-alpha activates signal transducer and activator of transcription-3 at reperfusion without involving classic prosurvival kinases (Akt and extracellular signal-regulated kinase). AB - BACKGROUND: We previously reported that tumor necrosis-factor-alpha (TNF-alpha) can mimic classic ischemic preconditioning (IPC) in a dose- and time-dependent manner. Because TNF-alpha activates the signal transducer and activator of transcription-3 (STAT-3), we hypothesized that TNF-alpha-induced preconditioning requires phosphorylation of STAT-3 rather than involving the classic prosurvival kinases, Akt and extracellular signal-regulated kinase (Erk) 1/2, during early reperfusion. METHODS AND RESULTS: Isolated, ischemic/reperfused rat hearts were preconditioned by either IPC or low-dose TNF-alpha (0.5 ng/mL). Western blot analysis confirmed that IPC phosphorylated Akt and Erk 1/2 after 5 minutes of reperfusion (Akt increased by 34+/-6% and Erk, by 105+/-28% versus control; P<0.01). Phosphatidylinositol 3-kinase/Akt inhibition (wortmannin) or mitogen activated protein kinase-Erk 1/2 kinase inhibition (PD-98059) during early reperfusion abolished the infarct-sparing effect of IPC. In contrast, TNF-alpha preconditioning did not phosphorylate these kinases (Akt increased by 7+/-7% and Erk, by 17+/-14% versus control; P=NS). Neither wortmannin nor PD-98059 inhibited TNF-alpha-mediated cardioprotection. However, TNF-alpha and IPC both phosphorylated STAT-3 and the proapoptotic protein Bcl-2 antagonist of cell death (BAD) (STAT-3 increased by 58+/-17% with TNF-alpha or by 68+/-12% with IPC; BAD increased by 75+/-8% with TNF-alpha or by 205+/-20% with IPC; P<0.01 versus control), thereby activating the former and inactivating the latter. The STAT-3 inhibitor AG 490 abolished cardioprotection and BAD phosphorylation with both preconditioning stimuli. CONCLUSIONS: Activation of the classic prosurvival kinases (Akt and Erk 1/2) is not essential for TNF-alpha-induced preconditioning in the early reperfusion phase. We show the existence of an alternative protective pathway that involves STAT-3 activation specifically at reperfusion in response to both TNF-alpha and classic IPC. This novel prosurvival pathway may have potential therapeutic significance. PMID- 16344383 TI - Metabolomic identification of novel biomarkers of myocardial ischemia. AB - BACKGROUND: Recognition of myocardial ischemia is critical both for the diagnosis of coronary artery disease and the selection and evaluation of therapy. Recent advances in proteomic and metabolic profiling technologies may offer the possibility of identifying novel biomarkers and pathways activated in myocardial ischemia. METHODS AND RESULTS: Blood samples were obtained before and after exercise stress testing from 36 patients, 18 of whom demonstrated inducible ischemia (cases) and 18 of whom did not (controls). Plasma was fractionated by liquid chromatography, and profiling of analytes was performed with a high sensitivity electrospray triple-quadrupole mass spectrometer under selected reaction monitoring conditions. Lactic acid and metabolites involved in skeletal muscle AMP catabolism increased after exercise in both cases and controls. In contrast, there was significant discordant regulation of multiple metabolites that either increased or decreased in cases but remained unchanged in controls. Functional pathway trend analysis with the use of novel software revealed that 6 members of the citric acid pathway were among the 23 most changed metabolites in cases (adjusted P=0.04). Furthermore, changes in 6 metabolites, including citric acid, differentiated cases from controls with a high degree of accuracy (P<0.0001; cross-validated c-statistic=0.83). CONCLUSIONS: We report the novel application of metabolomics to acute myocardial ischemia, in which we identified novel biomarkers of ischemia, and from pathway trend analysis, coordinate changes in groups of functionally related metabolites. PMID- 16344384 TI - Peristent remodeling and neointimal suppression 2 years after polymer-based, paclitaxel-eluting stent implantation: insights from serial intravascular ultrasound analysis in the TAXUS II study. AB - BACKGROUND: The purpose of this study was to evaluate long-term vascular responses as long as 2 years after implantation of polymer-based, paclitaxel eluting stents in contrast to uncoated stents. METHODS AND RESULTS: TAXUS II is a randomized, double-blind trial comparing slow-release (SR) and moderate-release (MR) TAXUS stents with bare-metal control stents (BMSs). One hundred sixty-one event-free patients (SR, 43; MR, 41; and BMS, 77) underwent serial intravascular ultrasound (IVUS) analysis after the procedure and at 6 months and 2 years. At 2 years, neointimal responses continued to be significantly suppressed in the SR and MR groups when compared with the BMS group (BMS, 1.49+/-1.12 mm2; SR, 0.94+/ 0.76 mm2 [P=0.004]; and MR, 1.06+/-0.90 mm2 [P=0.02]). Between 6 months and 2 years, the BMS group showed compaction of the neointima (Delta, -0.22+/-1.05 mm2 [P=0.08]). In contrast, both the SR and MR groups exhibited an increase (Delta SR, 0.30+/-0.76 mm2 (P=0.01); MR, 0.41+/-0.94 mm2 [P=0.009]). Between 6 months and 2 years, the initial increase in plaque outside the stent regressed in the BMS and SR groups to levels comparable to those after the procedure, whereas expansive remodeling partially regressed in the MR group (Delta between after the procedure and 2 years BMS, -0.34+/-1.28 mm2 [P=0.05]; SR, -0.02+/-1.40 mm2 [P=0.93]; MR, 0.32+/-1.56 mm2 [P=0.27]). CONCLUSIONS: The 2-year follow-up demonstrates that neointimal suppression was dose independent and that this effect was still sustained at 2 years. However, the increase in area outside the stent seen at 6 months regressed to different extents in a dose-dependent manner at 2 years. PMID- 16344385 TI - Twenty-year trends in serum cholesterol, hypercholesterolemia, and cholesterol medication use: the Minnesota Heart Survey, 1980-1982 to 2000-2002. AB - BACKGROUND: Although US cholesterol concentrations have dropped, &50% of adults have total cholesterol concentrations > or =5.18 mmol/L, putting them at "borderline-high risk" for heart disease. Whether the decline has continued into the 21st century is unknown. We assessed 20-year trends in cholesterol, hypercholesterolemia, lipid-lowering drug use, and cholesterol awareness, treatment, and control from Minnesota Heart Survey (MHS) data. METHODS AND RESULTS: Five independent, cross-sectional, population-based surveys of 2500 to 5000 adults were conducted in the Minneapolis-St. Paul, Minn, area from 1980 to 2002. Mean (nonfasting) total cholesterol concentrations have continued a 20-year decline, punctuated by an intervening lull. Age-adjusted mean total cholesterol concentrations in 2000 to 2002 were 5.16 and 5.09 mmol/L for men and women, respectively (in 1980 to 1982, 5.49 and 5.38 mmol/L for men and women, respectively) However, the decline has not been uniform across all age groups. Middle-aged to older people have shown substantial decreases, but younger people have shown little overall change and recently had increased total cholesterol values. The mean prevalence of hypercholesterolemia in 2000 to 2002 was 54.9% for men and 46.5% for women and has decreased significantly for both during the study. Age-adjusted mean high-density lipoprotein cholesterol concentrations in 2000 to 2002 were 1.09 and 1.40 mmol/L for men and women, respectively, and were not different from the prior survey. Lipid-lowering drug use rose significantly for both sexes aged 35 to 74 years. Awareness, treatment, and control of hypercholesterolemia have increased; however, more than half of those at borderline-high risk remain unaware of their condition. CONCLUSIONS: Although hypercholesterolemia prevalence continued to fall, significant population segments still have cholesterol concentrations near or at the level of increased risk. PMID- 16344386 TI - Blood flow remodels growing vasculature during vascular endothelial growth factor gene therapy and determines between capillary arterialization and sprouting angiogenesis. AB - BACKGROUND: For clinically relevant proangiogenic therapy, it would be essential that the growth of the whole vascular tree is promoted. Vascular endothelial growth factor (VEGF) is well known to induce angiogenesis, but its capability to promote growth of larger vessels is controversial. We hypothesized that blood flow remodels vascular growth during VEGF gene therapy and may contribute to the growth of large vessels. METHODS AND RESULTS: Adenoviral (Ad) VEGF or LacZ control gene transfer was performed in rabbit hindlimb semimembranous muscles with or without ligation of the profound femoral artery (PFA). Contrast-enhanced ultrasound and dynamic susceptibility contrast MRI demonstrated dramatic 23- to 27-fold increases in perfusion index and a strong decrease in peripheral resistance 6 days after AdVEGF gene transfer in normal muscles. Enlargement by 20 fold, increased pericyte coverage, and decreased alkaline phosphatase and dipeptidyl peptidase IV activities suggested the transformation of capillaries toward an arterial phenotype. Increase in muscle perfusion was attenuated, and blood vessel growth was more variable, showing more sprouting angiogenesis and formation of blood lacunae after AdVEGF gene transfer in muscles with ligated PFA than in normal muscles. Three-dimensional ultrasound reconstructions and histology showed that the whole vascular tree, including large arteries and veins, was enlarged manifold by AdVEGF. Blood flow was normalized and enlarged collaterals persisted in operated limbs 14 days after AdVEGF treatment. CONCLUSIONS: This study shows that (1) blood flow modulates vessel growth during VEGF gene therapy and (2) VEGF overexpression promotes growth of arteries and veins and induces capillary arterialization leading to supraphysiological blood flow in target muscles. PMID- 16344387 TI - Arrhythmogenic right ventricular dysplasia: a United States experience. AB - BACKGROUND: Arrhythmogenic right ventricular dysplasia (ARVD) is an inherited cardiomyopathy characterized by right ventricular dysfunction and ventricular arrhythmias. The purpose of our study was to describe the presentation, clinical features, survival, and natural history of ARVD in a large cohort of patients from the United States. METHODS AND RESULTS: The patient population included 100 ARVD patients (51 male; median age at presentation, 26 [interquartile range {IQR}, 18 to 38; range, 2 to 70] years). A familial pattern was observed in 32 patients. The most common presenting symptoms were palpitations, syncope, and sudden cardiac death (SCD) in 27%, 26%, and 23% of patients, respectively. Among those who were diagnosed while living (n=69), the median time between first presentation and diagnosis was 1 (range, 0 to 37) year. During a median follow-up of 6 (IQR, 2 to 13; range, 0 to 37) years, implantable cardioverter/defibrillators (ICD) were implanted in 47 patients, 29 of whom received an appropriate ICD discharge, including 3 patients who received the ICD for primary prevention. At follow-up, 66 patients were alive, of whom 44 had an ICD in place, 5 developed signs of heart failure, 2 had a heart transplant, and 18 were on drug therapy. Thirty-four patients died either at presentation (n=23: 21 SCD, 2 noncardiac deaths) or during follow-up (n=11: 10 SCD, 1 of biventricular heart failure), of whom only 3 were diagnosed while living and 1 had an ICD implanted. On Kaplan-Meier analysis, the median survival in the entire population was 60 years. CONCLUSIONS: ARVD patients present between the second and fifth decades of life either with symptoms of palpitations and syncope associated with ventricular tachycardia or with SCD. Diagnosis is often delayed. Once diagnosed and treated with an ICD, mortality is low. There is a wide variation in presentation and course of ARVD patients, which can likely be explained by the genetic heterogeneity of the disease. PMID- 16344390 TI - Cardiology patient page. Automated external defibrillators in the public domain: am I ready to use one? PMID- 16344391 TI - Images in cardiovascular medicine. Cardiac failure in the chick embryo resembles heart failure in humans. PMID- 16344392 TI - Images in cardiovascular medicine. Bedside diagnosis of cardiac rupture by contrast echocardiography. PMID- 16344393 TI - Images in cardiovascular medicine. Cast of complex congenital heart malformation in a living patient. PMID- 16344394 TI - Letter regarding article by Mauri et al, "Late loss in lumen diameter and binary restenosis for drug-eluting stent comparison". PMID- 16344395 TI - Letter regarding article by Rotmans et al, "In vivo cell seeding with anti-CD34 antibodies successfully accelerates endothelialization but stimulates intimal hyperplasia in porcine arteriovenous expanded polytetrafluoroethylene grafts". PMID- 16344396 TI - "La Donna e Mobile...": is cardiac neuronal nitric oxide synthase such a disconcerting enzyme? PMID- 16344397 TI - Structural heart disease, SCN5A gene mutations, and Brugada syndrome: a complex menage a trois. PMID- 16344398 TI - Critical questions about the metabolic syndrome. PMID- 16344399 TI - Virtual reality in interventional electrophysiology. PMID- 16344400 TI - Cardiac histological substrate in patients with clinical phenotype of Brugada syndrome. AB - BACKGROUND: The role of structural heart disease and sodium channel dysfunction in the induction of electrical instability in Brugada syndrome is still debated. METHODS AND RESULTS: We studied 18 consecutive patients (15 males, 3 females; mean age 42.0+/-12.4 years) with clinical phenotype of Brugada syndrome and normal cardiac structure and function on noninvasive examinations. Clinical presentation was ventricular fibrillation in 7 patients, sustained polymorphic ventricular tachycardia in 7, and syncope in 4. All patients underwent cardiac catheterization, coronary and ventricular angiography, biventricular endomyocardial biopsy, and DNA screening of the SCN5A gene. Biopsy samples were processed for histology, electron microscopy, and molecular screening for viral genomes. Microaneurysms were detected in the right ventricle in 7 patients and also in the left ventricle in 4 of them. Histology showed a prevalent or localized right ventricular myocarditis in 14 patients, with detectable viral genomes in 4; right ventricular cardiomyopathy in 1 patient; and cardiomyopathic changes in 3. Genetic studies identified 4 carriers of SCN5A gene mutations that cause in vitro abnormal function of mutant proteins. In these patients, myocyte cytoplasm degeneration was present at histology, whereas terminal dUTP nick end labeling assay showed a significant increase of apoptotic myocytes in right and left ventricle versus normal controls (P=0.014 and P=0.013, respectively). CONCLUSIONS: Despite an apparently normal heart at noninvasive evaluation, endomyocardial biopsy detected structural alterations in all 18 patients with Brugada syndrome. Mutations in the SCN5A gene, identified in 4 of the 18 patients, may have induced concealed structural abnormalities of myocardiocytes that accounted for paroxysmal arrhythmic manifestations. PMID- 16344401 TI - Techniques, evaluation, and consequences of linear block at the left atrial roof in paroxysmal atrial fibrillation: a prospective randomized study. AB - BACKGROUND: There are no reports describing the technique, electrophysiological evaluation, and clinical consequences of complete linear block at roofline joining the superior pulmonary veins (PVs) in patients with paroxysmal atrial fibrillation (AF). METHODS AND RESULTS: Ninety patients with drug-refractory paroxysmal AF undergoing radiofrequency ablation were prospectively randomized into 2 ablation strategies: (1) PV isolation (n=45) or (2) PV isolation in combination with linear ablation joining the 2 superior PVs (roofline; n=45). In both groups, the cavotricuspid isthmus, fragmented peri-PV-ostial electrograms, and spontaneous non-PV foci were ablated. Roofline ablation was performed at the most cranial part of the left atrium (LA) with complete conduction block demonstrated during LA appendage pacing by the online mapping of continuous double potential and an activation detour propagating around the PVs to activate caudocranially the posterior wall of the LA. The effect of ablation at the LA roof was evaluated by the change in fibrillatory cycle length, termination and noninducibility of AF, and clinical outcome. PV isolation was achieved in all patients with no significant differences in the radiofrequency duration, fluoroscopy, or procedural time between the groups. Roofline ablation required 12+/-6 (median 11, range 3 to 25) minutes of radiofrequency energy delivery with a fluoroscopic duration of 7+/-2 minutes and was performed in 19+/-7 minutes. Complete block was confirmed in 43 patients (96%) and resulted in an activation delay that was shorter circumventing the left than the right PVs during LA appendage pacing (138+/-15 versus 146+/-25 ms, respectively; P=0.01). Roofline ablation resulted in a significant increase in the fibrillatory cycle length (198+/-38 to 217+/-44 ms; P=0.0005), termination of arrhythmia in 47% (8/17), and subsequent noninducibility of AF in 59% (10/17) of the patients inducible after PV isolation. However, LA flutter, predominantly perimitral, could be induced in 10 patients (22%) after roofline ablation. At 15+/-4 months, 87% of the roofline group and 69% with PV isolation alone are arrhythmia free without antiarrhythmics (P=0.04). CONCLUSIONS: This prospective randomized study demonstrates the feasibility of achieving complete linear block at the LA roof. Such ablation resulted in the prolongation of the fibrillatory cycle, termination of AF, and subsequent noninducibility and is associated with an improved clinical outcome compared with PV isolation alone. PMID- 16344402 TI - Prediction of type 2 diabetes mellitus with alternative definitions of the metabolic syndrome: the Insulin Resistance Atherosclerosis Study. AB - BACKGROUND: In addition to predicting cardiovascular disease (CVD) morbidity and mortality, the metabolic syndrome is strongly associated with the development of type 2 diabetes mellitus (DM), itself an important risk factor for CVD. Our objective was to compare the ability of various metabolic syndrome criteria (including those recently proposed by the International Diabetes Federation), markers of insulin resistance (IR) and inflammation, and impaired glucose tolerance (IGT) in the prediction of DM and to determine whether various proposed modifications to the National Cholesterol Education program (NCEP) metabolic syndrome definition improved predictive ability. METHODS AND RESULTS: We examined 822 subjects in the Insulin Resistance Atherosclerosis Study aged 40 to 69 years who were nondiabetic at baseline. After 5.2 years, 148 individuals had developed DM. IGT, metabolic syndrome definitions, and IR markers all significantly predicted DM, with odds ratios ranging from 3.4 to 5.4 (all P<0.001), although there were no significant differences in the areas under the receiver operator characteristic (AROC) curves between the definitions. Modifying or requiring obesity, glucose, or IR components in NCEP-defined metabolic syndrome did not significantly alter the predictive ability of the definition under AROC curve criteria (all P>0.05). Similarly, although IR and inflammation variables were significantly associated with incident DM when included in multivariate models with NCEP-defined metabolic syndrome (all P<0.01), expanding the definition by adding these variables as components did not significantly alter the predictive ability of the definition under AROC curve criteria (all P>0.05). CONCLUSIONS: The International Diabetes Federation and NCEP metabolic syndrome definitions predicted DM at least as well as the World Health Organization definition, despite not requiring the use of oral glucose tolerance testing or measures of IR or microalbuminuria. Modifications or additions to the NCEP metabolic syndrome definition had limited impact on the prediction of DM. PMID- 16344403 TI - nNOS gene deletion exacerbates pathological left ventricular remodeling and functional deterioration after myocardial infarction. AB - BACKGROUND: The neuronal isoform of nitric oxide synthase (nNOS) has been implicated in the regulation of basal and beta-adrenergic inotropy in normal and chronically infarcted hearts. Furthermore, myocardial nNOS expression and activity increase in failing hearts, raising the possibility that nNOS may influence left ventricular (LV) remodeling progression and functional deterioration after myocardial infarction (MI). METHODS AND RESULTS: We compared LV remodeling at 1, 4, and 8 weeks after MI in nNOS-knockout mice (nNOS(-/-)) and their wild-type (WT) littermates matched for infarct size by using a highly accurate 3-dimensional echocardiographic technique. Basal LV hemodynamics and the inotropic response to dobutamine infusion (4 and 16 ng.g(-1).min(-1)) were also evaluated 8 weeks after MI. Sham-operated nNOS(-/-) mice showed enhanced basal LV contractility (P<0.03 versus WT, as evaluated by preload-recruitable stroke work) but an attenuated inotropic response to dobutamine infusion (P<0.01 versus WT). Both basal and beta-adrenergic LV relaxations were significantly impaired in nNOS(-/-) mice. Survival after MI did not differ between groups. However, nNOS(-/ ) mice developed a faster and more severe LV dilation compared with WT mice (P<0.05 for both end-systolic and end-diastolic volume indices). WT mice maintained a positive inotropic response to dobutamine 8 weeks after MI. In contrast, infarcted nNOS(-/-) mice responded to dobutamine with a dramatic fall in LV contractility (P<0.01 for preload-recruitable stroke work). CONCLUSIONS: nNOS plays a crucial role in preventing adverse LV remodeling and maintaining myocardial beta-adrenergic reserve after MI. Taken together, our findings suggest that upregulation of myocardial nNOS in infarcted hearts may be an important adaptive mechanism. PMID- 16344404 TI - Cost-effectiveness of fixed-dose combination of isosorbide dinitrate and hydralazine therapy for blacks with heart failure. AB - BACKGROUND: Fixed-dose combination of isosorbide dinitrate/hydralazine (ISDN/HYD) improved clinical outcomes in the African-American Heart Failure Trial (A-HeFT). We assessed the resource use, costs of care, and cost-effectiveness of ISDN/HYD therapy in the A-HeFT trial population. METHODS AND RESULTS: We obtained resource use data from A-HeFT, assigning costs through the use of US federal sources. Excluding indirect costs, we summarized the within-trial experience and modeled cost-effectiveness over extended time horizons, including a US societal lifetime reference case. During the mean trial follow-up of 12.8 months, the ISDN/HYD group incurred fewer heart failure-related hospitalizations (0.33 versus 0.47 per subject; P=0.002) and shorter mean hospital stays (6.7 versus 7.9 days; P=0.006). When study drug costs were excluded, both heart failure-related and total healthcare costs were lower in the ISDN/HYD group (mean per-subject heart failure related costs, 5997 dollars versus 9144 dollars; P=0.04; mean per-subject total healthcare costs, 15,384 dollars versus 19,728 dollars; P=0.03). With an average daily drug cost of 6.38 dollars, ISDN/HYD therapy was dominant (reduced costs and improved outcomes) over the trial duration. Assuming that no additional benefits accrue beyond the trial, we project the cost-effectiveness of ISDN/HYD therapy using heart failure-related costs to be 16,600 dollars/life-year at 2 years after enrollment, 37,100 dollars/life-year at 5 years, and 41,800 dollars/life-year over lifetime (reference case). CONCLUSIONS: ISDN/HYD therapy, previously shown to improve clinical outcomes, also reduced resource use and costs in A-HeFT, primarily because of a large reduction in hospitalizations. Long-term use of ISDN/HYD therapy should be associated with a favorable cost-effectiveness profile in this population. PMID- 16344405 TI - Registration of three-dimensional left atrial computed tomographic images with projection images obtained using fluoroscopy. AB - BACKGROUND: Anatomic structures such as the left atrium and the pulmonary veins (PVs) are not delineated by fluoroscopy because there is no contrast differentiation between them and the surrounding anatomy. Representation of an anatomic structure via a 3D model obtained from computed tomography (CT) imaging and subsequent projection of these images over the fluoroscopy system may help in navigation of the mapping and ablation catheter to the appropriate sites during electrophysiology procedures. METHODS AND RESULTS: In this feasibility study, in vitro experiments were performed with a plastic heart model (phantom) with 2 catheters or radiopaque platinum beads placed in the phantom at the time of CT imaging and fluoroscopy. Subsequently, 20 consecutive patients underwent contrast enhanced, ECG-gated CT scanning. Left atrial volumes were generated from the reconstructed data at &75% of the R-R interval during the cardiac cycle. Similarly, the superior vena cava and the coronary sinus were also reconstructed from these images. During the electrophysiology procedure, digital records (cine sequences) were obtained. Using predetermined algorithms, both the phantom model and the patients' 3D left atrial models derived from the CT were registered with projection images of fluoroscopy. Registration was performed with a transformation that linked the superior vena cava and the coronary sinus from the CT model with a catheter placed inside the coronary sinus via the superior vena cava. Registration was successfully accomplished with the plastic phantom and in all 20 patients. Registration accuracy was assessed in the phantom by assessing the overlapping beads seen both in the CT and the fluoroscopy images. The mean registration error was 1.4 mm (range 0.9 to 2.3 mm). Accuracy of the registered images was assessed in patients with recordings from a basket catheter placed sequentially in the superior PVs and by injecting contrast into the PVs to assess overlapping of contrast-filled PVs with the corresponding vessels on the registered images. The images could be calibrated quite accurately. Any rotational error, which was usually minor, could be corrected by rotating the images as needed. CONCLUSIONS: Registration of 3D models of the left atrium and PVs with fluoroscopic images of the same is feasible and could enable appropriate navigation and localization of the mapping and ablation catheter during procedures such as atrial fibrillation ablation. PMID- 16344406 TI - Nuclear factor-kappaB-mediated cell survival involves transcriptional silencing of the mitochondrial death gene BNIP3 in ventricular myocytes. AB - BACKGROUND: A survival role for the transcription factor nuclear factor-kappaB (NF-kappaB) in ventricular myocytes has been reported; however, the underlying mechanism is undefined. In this report we provide new mechanistic evidence that survival signals conferred by NF-kappaB impinge on the hypoxia-inducible death factor BNIP3. METHODS AND RESULTS: Activation of the NF-kappaB signaling pathway by IKKbeta in ventricular myocytes suppressed mitochondrial permeability transition pore (PTP) opening and cell death provoked by BNIP3. Expression of IKKbeta or p65 NF-kappaB suppressed basal and hypoxia-inducible BNIP3 gene activity. Deletion analysis of the BNIP3 promoter revealed the NF-kappaB elements to be crucial for inhibiting basal and inducible BNIP3 gene activity. Cells derived from p65(-/-)-deficient mice or ventricular myocytes rendered defective for NF-kappaB signaling with a nonphosphorylative IkappaB exhibited increased basal BNIP3 gene expression, mitochondrial PTP, and cell death. Genetic or functional ablation of the BNIP3 gene in NF-kappaB-defective myocytes rescued them from mitochondrial defects and cell death. CONCLUSIONS: The data provide new compelling evidence that NF-kappaB suppresses mitochondrial defects and cell death of ventricular myocytes through a mechanism that transcriptionally silences the death gene BNIP3. Collectively, our data provide new mechanistic insight into the mode by which NF-kappaB suppresses cell death and identify BNIP3 as a key transcriptional target for NF-kappaB-regulated expression in ventricular myocytes. PMID- 16344407 TI - Acute aortic syndromes. PMID- 16344408 TI - Peer review and professionalism at the Archives of Internal Medicine. PMID- 16344409 TI - Residency training in the modern era: the pipe dream of less time to learn more, care better, and be more professional. PMID- 16344410 TI - History forgotten is history relived: nosocomial infection control is also essential in the outpatient setting. PMID- 16344412 TI - Refusal of hydration and nutrition: irrelevance of the "artificial" vs "natural" distinction. PMID- 16344413 TI - The American public and the Terri Schiavo case. AB - An important question for physicians in the aftermath of the Terri Schiavo case is whether the effort of elected officials to intervene was a one-time anomaly or signals a future trend of elected officials being involved in cases where patients are in a vegetative or long-term comatose state. To try to answer this question, we used results from 12 national opinion surveys conducted in March and April 2005, when the Schiavo case was being debated. A review of these survey results showed that efforts by elected politicians to intervene in the Schiavo case were opposed by the majority of Americans. However, the public was more divided on the question of whether Schiavo's feeding tube should be removed. Opposition to removing Schiavo's feeding tube was associated with opposition to abortion. The results suggest that issues involved in cases like Schiavo's are not likely to disappear from the political agenda. PMID- 16344414 TI - The case for medical treatment in chronic stable coronary artery disease. PMID- 16344415 TI - Angioplasty is better than medical therapy for alleviating chronic angina pectoris. PMID- 16344416 TI - Burnout and internal medicine resident work-hour restrictions. AB - BACKGROUND: Burnout is very common in internal medicine residents. Effective July 2003, all residents were restricted to work less than an average of 80 hours per week and no more than 30 hours of continuous duty for patient care and educational obligations. We evaluated rates of burnout in internal medicine residents before and after the implementation of the new work-hour restriction. METHODS: University of Colorado Health Science Center internal medicine residents were surveyed in May 2003 and May 2004. The survey contained the Maslach Burnout Inventory, organized into 3 subscales (ie, emotional exhaustion, depersonalization, and personal accomplishment); the Primary Care Evaluation of Mental Disorders depression screen; and self-reported quality of care and education. RESULTS: The response rate was 87% (121 of 139 residents) and 74% (106 of 143 residents) in 2003 and 2004, respectively. Self-reported hours worked decreased from a mean of 74.6 to 67.1 (P = .003). In 2004, 13% fewer residents experienced high emotional exhaustion (42% vs 29%; P = .03). There was a trend toward fewer residents with high depersonalization (61% vs 55%; P = .13) and fewer residents with a positive depression screen (51% vs 41%; P = .11). Personal accomplishment did not change. The assessment of self-reported quality of care did not significantly change from 2003 to 2004. Residents reported attending fewer educational conferences per month (18.99 vs 15.56; P = .01). Overall residency satisfaction decreased 6 mm on a 100-mm visual analogue score (P = .02). CONCLUSIONS: Burnout continues to be a major problem. Reducing hours may be the first step to reduce burnout but may also affect education and quality of care. PMID- 16344417 TI - The effects of work-hour limitations on resident well-being, patient care, and education in an internal medicine residency program. AB - BACKGROUND: The Accreditation Council for Graduate Medical Education work-hour limitations (WHLs) were implemented in July 2003. Effects on resident well-being, patient care, and education are not well understood. We investigated these effects of WHLs. METHODS: Self-administered survey of internal medicine residents in a university-based residency program in Seattle, Wash. Part of this survey was identical to one completed at our institution in 2001, permitting comparison of burnout, career satisfaction, and depression before and after WHLs. We surveyed 161 internal medicine residents, with 118 respondents (response rate, 73%). We measured resident well-being using the Maslach Burnout Inventory, a validated screening questionnaire for depression, and a previously described questionnaire for career satisfaction. We developed questions about overall agreement with implementation of WHLs and effects on resident well-being, patient care, and education. RESULTS: Comparison with the 2001 survey demonstrated an increase in the proportion of residents satisfied with their career (66% to 80%; P = .02) and a decrease in the proportion meeting criteria for emotional exhaustion (53% to 40%; P = .05). Slightly more residents reported a negative effect of WHLs on patient care (37%) than they did a positive (29%) or a neutral (34%) effect, and more reported a negative effect on their education (47%) than they did a positive (32%) or a neutral (21%) effect. Overall, most residents (65%) approved of WHLs. CONCLUSIONS: Internal medicine residents approve of WHLs overall and report benefits to their well-being. However, they also report negative effects on patient care and resident education. PMID- 16344418 TI - Residents report on adverse events and their causes. AB - BACKGROUND: Resident physicians are frontline providers with a unique vantage point from which to comment on patient safety-related events. METHODS: We surveyed trainees at 2 teaching hospitals about experiences with adverse events (AEs), mistakes, and near misses, as well as the potential causes. RESULTS: Responses were obtained from 821 (57%) of 1440 eligible trainees. Analysis was restricted to 689 clinical trainees. More than half (55%) reported ever caring for a patient who had an AE. The most common types of AEs were procedural and medication related. More than two thirds of AEs were considered significant. Of the most recent AEs, 24% were attributed to mistakes. The most common reasons for mistakes, as perceived by residents, were excessive work hours (19%), inadequate supervision (20%), and problems with handoffs (15%). In the last week, 114 respondents (18%) reported having a patient with an AE; of these, 42 (37%) reported AEs involving a mistake for which they considered themselves responsible. In addition, 141 (23%) reported near-miss incidents in the last week for which they considered themselves responsible. In multivariate analyses, significant predictors of AEs in the last week were inpatient rotation, duty hours in the last week, and procedural specialty. Predictors of near-miss errors in the last week were inpatient rotation, days of fatigue in the last month, and postgraduate year 1 status. CONCLUSIONS: These findings support the perception that AEs are commonly encountered by physicians and often associated with errors. Causes of errors in teaching hospitals appear to be multifactorial, and a variety of measures are necessary to improve safety. Eliciting residents' perspectives is important because residents may perceive events, actions, and causal relationships that medical record reviewers or observers cannot. PMID- 16344419 TI - Trust and sources of health information: the impact of the Internet and its implications for health care providers: findings from the first Health Information National Trends Survey. AB - BACKGROUND: The context in which patients consume health information has changed dramatically with diffusion of the Internet, advances in telemedicine, and changes in media health coverage. The objective of this study was to provide nationally representative estimates for health-related uses of the Internet, level of trust in health information sources, and preferences for cancer information sources. METHODS: Data from the Health Information National Trends Survey were used. A total of 6369 persons 18 years or older were studied. The main outcome measures were online health activities, levels of trust, and source preference. RESULTS: Analyses indicated that 63.0% (95% confidence interval [CI], 61.7%-64.3%) of the US adult population in 2003 reported ever going online, with 63.7% (95% CI, 61.7%-65.8%) of the online population having looked for health information for themselves or others at least once in the previous 12 months. Despite newly available communication channels, physicians remained the most highly trusted information source to patients, with 62.4% (95% CI, 60.8%-64.0%) of adults expressing a lot of trust in their physicians. When asked where they preferred going for specific health information, 49.5% (95% CI, 48.1%-50.8%) reported wanting to go to their physicians first. When asked where they actually went, 48.6% (95% CI, 46.1%-51.0%) reported going online first, with only 10.9% (95% CI, 9.5%-12.3%) going to their physicians first. CONCLUSION: The Health Information National Trends Survey data portray a tectonic shift in the ways in which patients consume health and medical information, with more patients looking for information online before talking with their physicians. PMID- 16344420 TI - Overestimation of chronic disability among elderly persons. AB - BACKGROUND: Although there is no generally accepted definition for the term short term disability, chronic disability has been defined as disability lasting or expected to last at least 90 days according to a protocol that was established by the National Long-Term Care Survey. We evaluated the validity of the established protocol and determined the accuracy of prevalence estimates of chronic disability among elderly persons in the United States. METHODS: Chronic disability was ascertained during a comprehensive assessment using the established protocol. Participants were subsequently classified as having chronic disability (the gold standard) based on the presence of disability during consecutive monthly interviews immediately before or after the comprehensive assessment. RESULTS: Of the 552 participants, 120 (21.7%) met criteria for chronic disability according to the established protocol. Of these, 30 (25.0%) and 39 (32.5%) did not meet criteria according to the gold standard under assumptions that were favorable and unfavorable (ie, stringent) to the established protocol, respectively. Conversely, of the 95 participants (17.2%) who met the gold standard criteria for chronic disability according to the favorable strategy and the 89 (16.1%) who met the criteria according to the stringent strategy, 5 (5.3%) and 8 (9.0%), respectively, did not meet criteria for chronic disability according to the established protocol. Relative to the established estimate of 7.0 million, our projections yielded about 2.0 million fewer chronically disabled elderly Americans in 1999. CONCLUSION: Our results threaten the validity of the currently established protocol for ascertaining chronic disability and suggest that the burden of chronic disability among elderly Americans has been substantially overestimated. PMID- 16344421 TI - Disparities in diabetes care: impact of mental illness. AB - BACKGROUND: Emerging evidence indicates that patients with mental health conditions (MHCs) may receive less intensive medical care. Diabetes serves as a useful condition in which to test for MHC-related disparities in care. We examined whether quality measures for diabetes care are worse for patients with or without MHCs. METHODS: This national, cross-sectional study included 313 586 noninstitutionalized Veterans Health Administration patients with diabetes (identified from diagnostic codes and prescriptions) whose Veterans Health Administration facility transmitted laboratory data to a central database; 76 799 (25%) had MHCs (based on diagnostic codes for depressed mood, anxiety, psychosis, manic symptoms, substance use disorders, personality disorders, and other categories). National data from Veterans Health Administration records, Medicare claims, and a national survey were linked to characterize 1999 diabetes care. RESULTS: Failure to meet diabetes performance measures was more common in patients with MHCs: unadjusted odds ratio (95% confidence interval) was 1.24 (1.22-1.27) for no hemoglobin A(1c) testing, 1.25 (1.23-1.28) for no low-density lipoprotein cholesterol testing, 1.05 (1.03-1.07) for no eye examination, 1.32 (1.30-1.35) for poor glycemic control, and 1.17 (1.15-1.20) for poor lipemic control. Disparities persisted after case mix adjustment and were more pronounced with specific MHCs (psychotic, manic, substance use, and personality disorders). The percentage not meeting diabetes care standards increased with increasing number of MHCs. CONCLUSION: Patients with mental illness merit special attention in national diabetes quality improvement efforts. PMID- 16344422 TI - Outbreak of catheter-associated Klebsiella oxytoca and Enterobacter cloacae bloodstream infections in an oncology chemotherapy center. AB - BACKGROUND: In March 2004, the Chicago Department of Public Health was notified of a cluster of bloodstream infections with Klebsiella oxytoca and Enterobacter cloacae at a chemotherapy center. Our purpose was to identify the source of the outbreak and prevent further cases. METHODS: The investigation included 103 oncology patients seen at an outpatient oncology chemotherapy center in Chicago during the 16 days before its closure. The outbreak investigation included case identification, retrospective cohort study, review of medical records, microbiologic testing of blood specimens, environmental cultures, and pulsed field gel electrophoresis. The main outcome measure was infection with K oxytoca, E cloacae, or both, and the Mantel-Haenszel chi(2) test was used to assess risk of infection in relation to presence of central venous catheter. RESULTS: Among the 103 patients, risk of infection was associated with the presence of central venous catheter (relative risk undefined, P<.001). Twenty-seven patients had blood cultures that grew K oxytoca, E cloacae, or both, and all had central venous catheters that were flushed with isotonic sodium chloride solution at the clinic from February 17 through March 3, 2004. Isolates of K oxytoca and E cloacae were matched by pulsed-field gel electrophoresis to K oxytoca and E cloacae isolates obtained from multiple predrawn syringes and from the intravenous fluid and administration set in use in the clinic at the time of its closing. CONCLUSIONS: The injection of contaminated isotonic sodium chloride solution through the venous catheters of attendees at the clinic likely provided the opportunity for bloodstream infections in these 27 case patients. This outbreak highlights the need for continued emphasis on safe injection practices and suggests the need for guidelines and recommendations tailored to outpatient settings. PMID- 16344423 TI - Metabolic syndrome vs Framingham Risk Score for prediction of coronary heart disease, stroke, and type 2 diabetes mellitus. AB - BACKGROUND: We sought to compare metabolic syndrome (MetS) with the Framingham Risk Score (FRS) as predictors of coronary heart disease (CHD), stroke, and type 2 diabetes mellitus (DM2) in middle-aged men. METHODS: A prospective study of 5128 men aged 40 to 59 years with no history of cardiovascular disease (CVD) (CHD or stroke) or DM2 drawn from general practices in 24 British towns and observed for 20 years. Metabolic syndrome was defined as the presence of 3 or more metabolic abnormalities based on modified National Cholesterol Education Program criteria. RESULTS: Men with MetS at baseline (26%) showed significantly higher relative risk (RR) than men without MetS of developing CHD (RR, 1.64; 95% confidence interval [CI], 1.41-1.90), stroke (RR, 1.61 95% CI, 1.26-2.06), and DM2 (RR, 3.57; 95% CI, 2.83-4.50). The probability of developing CVD or DM2 over 20 years increased from 11.9% in those with no abnormalities to 31.2% in those with 3 abnormalities to 40.8% in those with 4 or 5 abnormalities. The FRS was a better predictor of CHD and stroke than MetS but was less predictive of DM2. Areas under the receiver-operating characteristic curves for FRS vs the number of metabolic abnormalities were 0.68 vs 0.59 for CHD, 0.60 vs 0.70 for DM2, and 0.66 vs 0.55 for stroke (P< .001 for all). CONCLUSIONS: Presence of MetS is a significant predictor of CVD and DM2 but is a stronger predictor of DM2 than of CHD. Although MetS does not predict CHD as well as the FRS, it serves well as a simple clinical tool for identifying high-risk subjects predisposed to CVD or DM2. PMID- 16344424 TI - Long-term outcome of 231 patients with essential thrombocythemia: prognostic factors for thrombosis, bleeding, myelofibrosis, and leukemia. AB - BACKGROUND: Essential thrombocythemia (ET) is a clonal myeloproliferative disease associated with thrombohemorrhagic complications and myeloid transformation to diseases such as myelofibrosis and acute myeloid leukemia. METHODS: A multicenter study was conducted among 231 consecutive Chinese patients with ET. The literature about leukemogenic risk associated with the use of hydroxyurea therapy was reviewed. RESULTS: The median patient age was 65 years. Thrombosis rates at and after diagnosis of ET were comparable to those of white patients, but bleeding rates at and after diagnosis were much lower. The projected 10-year thrombosis-free, bleeding-free, and overall survival rates were 66%, 83%, and 80%, respectively. There were no deaths among patients 60 years or younger during a maximum follow-up of 15 years, and splenomegaly at diagnosis of ET appeared to protect against thrombosis. In multivariate analysis, advanced age predicted inferior 10-year thrombosis-free and overall survival, and male sex predicted inferior bleeding-free survival. Half the deaths were related to ET. The probability of myelofibrosis transformation was 9.7% at 10 years. Prior myelofibrosis (P = .008) and the use of melphalan treatment (P = .002) were risk factors for acute myeloid leukemia evolution. CONCLUSIONS: Essential thrombocythemia is a benign disease of older persons. Chinese patients have a low risk of bleeding, and prior myelofibrosis is a major risk factor for evolution to acute myeloid leukemia. Leukemic transformation with hydroxyurea therapy alone is rare and warrants further prospective studies. PMID- 16344425 TI - Renal function and risk of myocardial infarction in an elderly population: the Rotterdam Study. AB - BACKGROUND: Renal insufficiency is a risk factor for cardiovascular disease in patients with renal disease or coronary heart disease; however, it is unknown whether renal function is an independent predictor of cardiovascular disease in the general population. METHODS: We investigated whether the level of renal function, estimated by glomerular filtration rate, was associated with the risk of incident myocardial infarction among 4484 apparently healthy subjects in the Rotterdam Study (mean age, 69.6 years). We estimated the glomerular filtration rate by Cockcroft-Gault and abbreviated modification of diet in renal disease equations and used Cox regression analysis to estimate hazard ratios adjusted for cardiovascular risk factors, atherosclerosis, and medication use. RESULTS: During the follow-up period (mean, 8.6 years), 218 subjects (4.9%) had a myocardial infarction. A 10 mL/min per 1.73 m(2) decrease in glomerular filtration rate was associated with a 32% increased risk of myocardial infarction (P<.001). Compared with subjects in the fourth quartile, the multivariate-adjusted hazard ratios for the risk of myocardial infarction increased from 1.64 (95% confidence interval [CI], 1.03-2.59) in the third quartile to 1.94 (95% CI, 1.21-3.10) in the second quartile and 3.06 (95% CI, 1.80-5.19) in the quartile with the lowest glomerular filtration rate estimated by the Cockcroft-Gault equation. Using the abbreviated modification of diet in renal disease equation, the risk estimates for the third to first quartiles were 1.34 (95% CI, 0.89-2.01), 1.66 (95% CI, 1.14-2.49), and 1.90 (95% CI, 1.25-2.90), respectively. CONCLUSIONS: The present study shows that renal function is a graded and independent predictor of the development of myocardial infarction in an elderly population. Early detection of decreased renal function may identify subjects who are at heightened risk of coronary heart disease. PMID- 16344426 TI - Cystatin C and incident peripheral arterial disease events in the elderly: results from the Cardiovascular Health Study. AB - BACKGROUND: The association of cystatin C, a novel marker of renal function, with risk for developing complications related to peripheral arterial disease (PAD) has not been examined. METHODS: We evaluated the hypothesis that a high cystatin C concentration is independently associated with future PAD events among 4025 participants in the Cardiovascular Health Study who underwent serum cystatin C measurement at the 1992-1993 visit and who did not have PAD at baseline. The association of cystatin C quintiles with time to first lower-extremity PAD procedure (bypass surgery, angioplasty, or amputation) was evaluated using multivariable proportional hazards models. Secondary analyses were conducted using quintiles of serum creatinine level and estimated glomerular filtration rate (eGFR). RESULTS: The annualized risk of undergoing a procedure for PAD was 0.43% per year among participants in the highest cystatin C quintile (>1.27 mg/L) compared with 0.21% per year or less in all other quintiles. After multivariable adjustment for known risk factors for PAD, elevated cystatin C levels remained associated with the outcome (hazard ratio, 2.5 for highest vs lowest quintile of cystatin C, 95% confidence interval, 1.2-5.1). The highest quintiles of serum creatinine level and eGFR were not associated with future PAD events in either unadjusted or adjusted analyses. CONCLUSION: Elevated concentrations of cystatin C were independently predictive of incident PAD events among community-dwelling elderly patients. PMID- 16344427 TI - Outcomes in 45 patients with statin-associated myopathy. AB - BACKGROUND: Published studies regarding statin-associated myopathy may describe more dramatic patient presentations, potentially leading to inaccurate characterization of the condition. Furthermore, long-term outcomes and responses to statin rechallenge in patients with statin-associated myopathy are largely unknown. METHODS: The University of Wisconsin Hospital and Clinics Medical Informatics Department identified 437 patients with International Classification of Diseases, Ninth Revision codes potentially representing cases of statin associated myopathy from more than 13 years of inpatient and outpatient data; 45 of these individuals were diagnosed as having statin-associated myopathy. Using a standardized form, 2 researchers abstracted all the case records to define the clinical course of statin-induced myopathy. RESULTS: The mean (SD) duration of statin therapy before symptom onset was 6.3 (9.8) months. Resolution of muscle pain occurred a mean (SD) of 2.3 (3.0) months after discontinuation of statin therapy. Six patients (13%) were hospitalized for the management of rhabdomyolysis; 2 had reversible renal dysfunction, and 1 with preexisting renal insufficiency subsequently began lifelong dialysis. Hospitalized patients developed myopathy more quickly after initiating statin therapy (1.3 vs 7.1 months; P = .048) and were more likely to be taking concomitant medications known to increase the risk of statin-associated myopathy (P = .03). Thirty-seven patients received another statin after an episode of statin-associated myopathy; 21 (57%) reported recurrent muscle pain, whereas 16 (43%) tolerated other statins without recurrent symptoms. CONCLUSIONS: Patients with statin-associated myopathy experienced full resolution of muscle pain on cessation of statin therapy. Although no deaths occurred, 13% of the patients required hospitalization for rhabdomyolysis. Recurrent muscle pain was common on statin rechallenge. PMID- 16344428 TI - Venous thromboembolism among elderly patients treated with atypical and conventional antipsychotic agents. AB - BACKGROUND: Some antipsychotic agents have been indicated as a possible risk factor for venous thromboembolism (VTE) in adult patients with psychiatric disorders. The aim of this study was to estimate the effect of atypical and conventional antipsychotic agents on the risk of hospitalization for VTE among elderly patients. METHODS: We conducted a retrospective cohort study on nursing home residents in 5 states. We used data from the Minimum Data Set to identify 19 940 new users of antipsychotic agents and 112 078 nonusers. Hospitalization with VTE as primary discharge diagnosis was determined during a 6-month follow-up period using Medicare inpatient claims. Cox proportional hazards models provided estimates of effect adjusted for confounders. RESULTS: The rate of hospitalization for VTE was 0.91 per 100 person-years. Venous thrombosis accounted for 77.6% of events and 22.4% were pulmonary embolisms. Relative to nonusers, the rate of hospitalization for VTE was increased for users of atypical antipsychotic agents, including risperidone (adjusted hazard ratio [HR], 1.98; 95% confidence interval [CI], 1.40-2.78), olanzapine (adjusted HR, 1.87; 95% CI, 1.06-3.27), and clozapine and quetiapine fumarate (adjusted HR, 2.68; 95% CI, 1.15-6.28). No increased rate was associated with phenothiazines (adjusted HR, 1.03; 95% CI, 0.60-1.77) or other conventional agents (adjusted HR, 0.98; 95% CI, 0.52-1.87). CONCLUSIONS: Atypical antipsychotic agents appear to increase the risk of VTE. However, these events are rare, and in clinical practice the absolute risk should be weighed against the effectiveness of these medications in the elderly population. PMID- 16344429 TI - Tea consumption and ovarian cancer risk in a population-based cohort. AB - BACKGROUND: Substantial evidence from laboratory studies indicates that green and black tea preparations may protect against various cancers. Few epidemiologic studies, however, have examined the relationship specifically between tea consumption and risk of ovarian cancer. METHODS: We prospectively examined the association between tea consumption and risk of ovarian cancer in 61 057 women aged 40 to 76 years who were participants in the population-based Swedish Mammography Cohort. Participants completed a validated 67-item food frequency questionnaire at enrollment between 1987 and 1990 and were followed for cancer incidence through December 2004. RESULTS: During an average follow-up of 15.1 years, 301 incident cases of invasive epithelial ovarian cancer were ascertained. Tea consumption was inversely associated with the risk of ovarian cancer after controlling for potential confounders (P for trend, .03). Compared with women who never or seldom (less than monthly) consumed tea, the multivariate hazard ratios for those who consumed less than 1 cup per day, 1 cup per day, and 2 or more cups per day were 0.82 (95% confidence interval [CI], 0.62-1.08), 0.76 (95% CI, 0.56 1.04), and 0.54 (95% CI, 0.31-0.91), respectively. Each additional cup of tea per day was associated with an 18% lower risk of ovarian cancer (multivariate hazard ratio, 0.82; 95% CI, 0.68-0.99). CONCLUSION: These results suggest that tea consumption is associated with a reduced risk of epithelial ovarian cancer in a dose-response manner. PMID- 16344430 TI - Preventing venous thromboembolism, especially in elderly people. PMID- 16344434 TI - Treatment of retinal angiomatous proliferation in age-related macular degeneration: a series of 104 cases of retinal angiomatous proliferation. AB - OBJECTIVE: To report the management of retinal angiomatous proliferation (RAP), a recently described intraretinal neovascular lesion occurring in age-related macular degeneration. METHODS: This was a retrospective review of consecutive patients with age-related macular degeneration who underwent treatment of RAP from January 1, 2000, through January 31, 2003. Inclusion criteria were age 55 years or older, signs of age-related macular degeneration, and diagnosis of RAP based on dynamic indocyanine green angiography. Baseline angiograms were reviewed and RAP was classified into the following 3 stages: stage 1, intraretinal neovascularization, early stage; stage 2, subretinal neovascularization, middle stage; and stage 3, choroidal neovascularization, late stage. Treatment and concomitant treatment results were assessed separately for each RAP stage. The clinical data were statistically analyzed (chi2 test and analysis of variance) for 2 main outcome measures--complete obliteration of the lesion and final visual acuity. RESULTS: Eighty-one patients (99 eyes) with 104 RAPs were identified. Forty-two lesions were at stage 1, 42 at stage 2, and 20 at stage 3. The following 5 treatments were performed: direct laser photocoagulation of the vascular lesion, laser photocoagulation of the feeder retinal arteriole, scatter "gridlike" laser photocoagulation, photodynamic therapy, and transpupillary thermotherapy. Complete obliteration of RAP was achieved in about 24 (57.1%) of the stage 1 lesions (direct laser photocoagulation of the vascular lesion, 73% success rate; photodynamic therapy, 45%), 11 (26.2%) of the stage 2 lesions (scatter gridlike laser photocoagulation, 38% success rate; direct laser photocoagulation of the vascular lesion, 17%), and only 3 (15.0%) of stage 3 lesions (P = .001). Predictive factors with a significant effect on final visual acuity were initial visual acuity (P = .003) and early lesion stage (P = .04). Best final visual acuity was 0.41 (mean, direct laser photocoagulation of the vascular lesion in stage 1) and 0.39 (mean, photodynamic therapy in stage 1), with a mean decrease of 2.5 and 3 lines from baseline, respectively. CONCLUSIONS: Treatment of RAP remains difficult. Early detection of the lesion and subsequent direct conventional laser photocoagulation seems to be associated with better anatomical and functional outcome. Once the vascular complex is well established, anatomical closure is rarely achieved. Further study is warranted to assess the long-term efficacy and the need for re-treatment. PMID- 16344433 TI - Development of metastatic disease after enrollment in the COMS trials for treatment of choroidal melanoma: Collaborative Ocular Melanoma Study Group Report No. 26. AB - OBJECTIVE: To describe the time between treatment for choroidal melanoma and first diagnosis of metastatic disease, sites of metastasis, treatments for metastasis, and time between diagnosis of metastasis and death. DESIGN: Prospective, longitudinal follow-up of patients diagnosed with choroidal melanoma who were enrolled in 2 randomized trials conducted by the Collaborative Ocular Melanoma Study Group. METHODS: Systemic and laboratory evaluations were performed during follow-up according to a standard protocol for 2320 patients enrolled in the Collaborative Ocular Melanoma Study trials without evidence of melanoma metastasis or other primary cancer at baseline. RESULTS: Seven hundred thirty nine patients were diagnosed with at least 1 site of metastasis during follow-up after treatment for choroidal melanoma. Five- and 10-year cumulative metastasis rates were 25% (95% confidence interval, 23%-27%) and 34% (95% confidence interval, 32%-37%), respectively. Liver was the most common site (89%). The death rate following the report of melanoma metastasis was 80% at 1 year (95% confidence interval, 77%-83%) and 92% at 2 years (95% confidence interval, 89% 94%). Overall survival after metastasis did not vary by baseline size of primary tumor nor treatment for metastasis (when known). Long-term survival after diagnosis of metastasis was uncommon; only 8 patients survived 5 or more years. CONCLUSION: Metastasis rate increased significantly with increasing primary tumor dimensions at time of patient enrollment. Prognosis after metastatic disease remains poor. Effective methods are needed to prevent, diagnose, and treat metastasis from choroidal melanoma. PMID- 16344435 TI - Analysis of the macula with optical coherence tomography after successful surgery for proliferative vitreoretinopathy. AB - OBJECTIVE: To use optical coherence tomography to assess the in vivo pathologic findings associated with incomplete visual recovery in patients who have undergone anatomically successful surgery to treat proliferative vitreoretinopathy. METHODS: Eligible patients were recruited in vitreoretinal outpatient clinics between April 1, 2002, and July 31, 2003. Patients were included who had undergone anatomically successful vitreoretinal surgery to treat proliferative vitreoretinopathy and, at least 3 months after surgery, had postoperative vision worse than expected (< or =6/12) with no identifiable cause at clinical examination. Patients underwent optical coherence tomography, stereo fundus fluorescein angiography was performed in a cohort of patients, and angiographic findings were compared with those on the optical coherence tomograms. Relevant clinical data were collected retrospectively from patient case notes. RESULTS: A total of 35 patients were recruited. Optical coherence tomograms revealed cystoid macular edema in 23 patients (66%) but did not identify any other specific intraretinal disease. Location of edema (outer or inner retina), determined with stereo fundus fluorescein angiography and optical coherence tomography, correlated well. CONCLUSIONS: Optical coherence tomography is a useful diagnostic tool for assessing poor postoperative visual acuity and can reveal disease undetected at clinical examination. Cystoid macular edema is a common finding on optical coherence tomograms in eyes with incomplete visual recovery after anatomically successful surgery to treat proliferative vitreoretinopathy. PMID- 16344436 TI - Factors predictive of tumor growth, tumor decalcification, choroidal neovascularization, and visual outcome in 74 eyes with choroidal osteoma. AB - OBJECTIVE: To evaluate choroidal osteoma for tumor growth, tumor decalcification, related choroidal neovascularization, visual acuity loss, and poor visual acuity. DESIGN: Retrospective nonrandomized single-center case series. SETTING: Ocular Oncology Service at Wills Eye Hospital, Thomas Jefferson University, Philadelphia, Pa. PARTICIPANTS: There were 74 eyes of 61 patients with choroidal osteoma evaluated between January 1, 1977, and January 1, 2003. MAIN OUTCOME MEASURES: The 5 outcome measures included tumor growth, tumor decalcification, related choroidal neovascularization, visual acuity loss of 3 or more Snellen lines, and poor visual acuity of 20/200 or worse. RESULTS: At 5 and 10 years, Kaplan-Meier analysis revealed tumor growth in 22% and 51% of eyes, tumor decalcification in 28% and 46% of eyes, choroidal neovascularization in 31% and 31% of eyes, visual acuity loss in 26% and 45% of eyes, and poor visual acuity in 45% and 56% of eyes, respectively. The clinical factor predictive of tumor growth was absent overlying retinal pigment epithelial alterations. The factor predictive of decalcification was irregular tumor surface. Of the 15 tumors that showed partial decalcification at the first visit, there was no further tumor growth in any case. Of the remaining 12 tumors that later developed decalcification, tumor growth, if it occurred, was along the margin opposite the decalcification. No tumor showed growth in the region of decalcification. Factors predictive of choroidal neovascularization included irregular tumor surface and subretinal hemorrhage. In 6 eyes that had both choroidal neovascularization and tumor decalcification, the neovascularization was detected prior to or at the same time as the decalcification. The factor predictive of visual acuity loss was presence of subretinal fluid whereas the factors predictive of poor visual acuity included symptoms and tumor decalcification. A comparison of eyes with subfoveal vs extrafoveal choroidal osteoma showed poor visual acuity in 15 (34%) of 44 eyes and 3 (10%) of 30 eyes, respectively. Eyes with decalcified choroidal osteomas manifested poor visual acuity in 13 (48%) of 27 eyes whereas those with nondecalcified tumors showed poor visual acuity in 5 (11%) of 47 eyes. CONCLUSIONS: Choroidal osteoma showed evidence of growth in 51% of eyes and decalcification in nearly 50% of eyes by 10 years. Tumors with any degree of decalcification at the initial visit showed no further growth. Overall, poor visual acuity of 20/200 or worse was found in 56% of eyes by 10 years, and decalcified subfoveal choroidal osteomas displayed a particularly poor visual prognosis. PMID- 16344437 TI - Blepharokeratoconjunctivitis in children. AB - OBJECTIVE: To evaluate the incidence, history, symptoms, clinical signs, and treatment outcomes of blepharokeratoconjunctivitis in a pediatric population at a tertiary cornea practice. METHODS: In a retrospective case series, we reviewed the medical records of all new pediatric patients from January 1, 1997, through December 31, 2002, noting the reason for referral and subsequent diagnosis. We further noted the history, clinical characteristics, and treatment outcomes of the patients with blepharokeratoconjunctivitis. RESULTS: Review of 195 medical records revealed that blepharokeratoconjunctivitis was the most common single diagnosis at consultation, accounting for 15% of referrals. Of the 29 cases identified, there were 16 girls (55%) and 13 boys (45%). The mean age at consultation was 6(1/2) years (age range, 2-12 years). On initial ophthalmologic examination, 11 (38%) of 29 patients were taking full-strength steroids and 4 patients (14%) were taking oral erythromycin. Oral therapy, in the form of erythromycin (n = 21) and doxycycline (n = 1), was prescribed to most patients (22/29 [76%]). Therapy with topical steroids was tapered at the initial visit in all patients. Follow-up was available for 15 of 29 patients, with a mean follow up of 5.4 months (range, 2-25 months). The condition of all patients showed clinical improvement. Recurrences were noted in 6 (40%) or 15 patients; all were successfully managed with low-potency steroid therapy. CONCLUSIONS: Blepharokeratoconjunctivitis is a common reason for cornea referral in children. Oral erythromycin therapy is an effective treatment with a steroid-sparing effect. Recurrences are common and may be successfully managed with low-potency steroid therapy. PMID- 16344438 TI - Minimally invasive orbital decompression: local anesthesia and hand-carved bone. AB - OBJECTIVE: To investigate the safety and efficacy of a conservative orbital decompression using sharp-curette bony decompression and intraconal fat debulking through a transconjunctival incision in patients with thyroid-related orbitopathy and mild to moderate proptosis. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS AND METHODS: Data from all patients undergoing minimal orbital decompression at the Jules Stein Eye Institute, Los Angeles, Calif, over a period of 4(1/4) years were collected and analyzed. Data included visual acuity, exophthalmometry measurements, intraocular pressure, complete slitlamp examination results, ocular ductions, new-onset primary or downgaze diplopia, and patient satisfaction. Conservative decompression was performed through a transconjunctival incision using a manual curette and by removing cortical bone from the zygomatic marrow space on the anterior rim of the inferior orbital fissure; intraconal fat was bluntly dissected and excised or suctioned with a Frasier tip aspirator. MAIN OUTCOME MEASURES: Patient perception of pressure pain and ocular discomfort, proptosis, visual acuity, intraocular pressure, postoperative complications, and new-onset primary or downgaze diplopia. RESULTS: Eighty minimally invasive orbital decompression surgeries were performed in 48 patients (6 male, 42 female). Six surgeries (4 patients) were performed for prominent globes with relative proptosis and no thyroid-related orbitopathy (non-Graves proptosis). All patients had improvement in congestive orbitopathy and pressure pain associated with thyroid-related orbitopathy. Exophthalmos decreased by a mean +/- SD of 2.4 +/- 2.6 mm from 22.7 +/- 2.5 mm (range, 17-29 mm) to 20.3 +/- 2.3 mm (range, 14-25 mm) (P<.001 [95% confidence interval, 1.8-3.0]). Mean visual acuity improved after surgery (P = .02). One patient (2.1%) developed postoperative primary or downgaze diplopia; he underwent successful eye muscle surgery at a later stage. No complications were associated with orbital decompression. CONCLUSIONS: Minimally invasive orbital decompression surgery with intraconal fat debulking in this group of patients was effective in proptosis reduction; improvement in subjective pressure pain and high patient satisfaction were noticed. Surgery was associated with a low rate (2.1%) of new onset primary or downgaze diplopia. Proptosis reduction using a graded approach accounting for 4 mm of retrodisplacement was achieved. PMID- 16344439 TI - Technetium tc 99m-labeled red blood cells in the preoperative diagnosis of cavernous hemangioma and other vascular orbital tumors. AB - OBJECTIVES: To evaluate technetium Tc 99m (99mTc) red blood cell scintigraphy as a diagnostic tool for orbital cavernous hemangioma and to differentiate between orbital masses on the basis of their vascularization. METHODS: We performed 99mTc red blood cell scintigraphy on 23 patients (8 female and 15 male; mean age, 47 years) affected by an orbital mass previously revealed with computed tomography (CT) and magnetic resonance imaging (MRI) and suggesting cavernous hemangioma. In our diagnosis, we considered the orbital increase delayed uptake with the typical scintigraphic pattern known as perfusion blood pool mismatch. The patients underwent biopsy or surgical treatment with transconjunctival cryosurgical extraction when possible. RESULTS: Single-photon emission tomography (SPET) showed intense focal uptake in the orbit corresponding to radiologic findings in 11 patients who underwent surgical treatment and pathologic evaluation (9 cavernous hemangiomas, 1 hemangiopericytoma, and 1 lymphangioma). Clinical or histologic examination of the remaining 22 patients revealed the presence of 5 lymphoid pseudotumors, 2 lymphomas, 2 pleomorphic adenomas of the lacrimal gland, 1 astrocytoma, 1 ophthalmic vein thrombosis, and 1 orbital varix. CONCLUSIONS: The confirmation of the preoperative diagnosis by 99mTc red blood cell scintigraphy shows that this technique is a reliable tool for differentiating cavernous hemangiomas from other orbital masses (sensitivity, 100%; specificity, 86%) when ultrasound, CT, and MRI are not diagnostic. Unfortunately, 99mTc red blood cell scintigraphy results were positive in 1 patient with hemangiopericytoma and 1 patient with lymphangioma, which showed increased uptake in the lesion on SPET images because of the vascular nature of these tumors. Therefore, in these cases, the SPET images have to be integrated with data regarding clinical preoperative evaluation and CT scans or MRI studies. On the basis of our study, a complete diagnostic picture, CT scans or MRI studies, and scintigraphic patterns can establish the preoperative diagnosis of vascular orbital tumors such as cavernous hemangioma, adult-type lymphangioma, and hemangiopericytoma. PMID- 16344440 TI - Effect of hypoxemia and hyperglycemia on pH in the intact cat retina. AB - OBJECTIVE: To examine the effects of acute hypoxemia and hyperglycemia on retinal pH to understand hyperglycemia-induced changes in the normal intact cat retina. METHODS: Spatial profiles of extracellular hydrogen ion (H+) concentration were obtained from the cat retina, in vivo, using pH-sensitive microelectrodes during normoxia (arterial partial pressure of oxygen [PaO2] = 114.5 +/- 7.9 mm Hg), normoglycemia (plasma glucose concentration, 117 +/- 19 mg/dL), acute hypoxemia (PaO2 = 29.5 +/- 2.2 mm Hg), and acute hyperglycemia (plasma glucose concentration, 303 +/- 67 mg/dL). An H+ diffusion model was fitted to the outer retinal data to quantify photoreceptor H+ production. The inner retinal pH was also examined. RESULTS: Hypoxemia induced a mean acute panretinal acidification of 0.16 pH units that originated from a 2.55-fold increase in net photoreceptor H+ production. Hyperglycemia induced an acute panretinal acidification of 0.12 pH units; however, photoreceptor H+ production levels remained unchanged. Retinal pH changes followed the course of arterial PaO2 and blood glucose changes. CONCLUSIONS: The increase in photoreceptor H+ production during hypoxemia confirms the importance of glycolysis in the retina. Hyperglycemia-induced pH changes resulted from either increased inner retinal H+ production or decreased H+ clearance/neutralization. Clinical Relevance The hyperglycemia-induced acidification that originates in the inner retina suggests that retinal acidosis may contribute to the development of diabetic retinal disease. PMID- 16344441 TI - Racial differences in glaucoma care: the longitudinal pattern of care. AB - OBJECTIVE: To examine if racial differences exist in longitudinal care patterns for Medicare beneficiaries with glaucoma. METHODS: We analyzed national longitudinal Medicare claims data from January 1, 1991, through December 31, 1999 in 21 644 Medicare beneficiaries linked to the National Long-Term Care Survey. Logistic regression was used to predict whether a person underwent an eye examination or eye surgery during the year, and negative binomial regression was used to predict the number of eye examinations and surgical procedures for glaucoma per year. Annual use of eye examinations was nearly identical for black persons (1.85 per year) and white persons (1.89 per year), whereas surgery rates were higher among blacks (0.15) than whites (0.08, P<.001). RESULTS: Blacks were more likely than whites to have glaucoma diagnosed, but rates among whites were higher than in prior population-based studies. When we controlled for other factors, blacks were not significantly less likely to undergo eye examination during the year; however, blacks were 78% more likely to undergo surgery (P < .001) and had 76% higher rates of surgical procedures (P < .001). CONCLUSIONS: No systematic pattern was found of underuse among blacks relative to whites after glaucoma diagnosis. Higher rates of surgery among blacks may indicate delayed onset of care and/or greater disease severity. PMID- 16344442 TI - Visual impairment and eye abnormalities in Oklahoma Indians. AB - OBJECTIVE: To determine the prevalence of visual impairment and eye abnormalities in Oklahoma Indians. METHODS: The cross-sectional study included 1019 Oklahoma Indians, aged 48 to 82 years; 60.2% were women. All participants gave a personal interview, and all underwent an eye examination, including the determination of best-corrected visual acuity and an ophthalmoscopic examination. In addition, two 45 degrees fundus photographs were taken of each eye, and these photographs were graded by the Fundus Photography Reading Center at the University of Wisconsin, Madison. RESULTS: Among the 1019 participants, 77.4% had a visual acuity of 20/20 or better, 19.5% and 2.5% had visual acuities of between 20/25 and 20/40 and between 20/50 and 20/190, respectively; and 0.6% were legally blind, all in the better eye. Cataract was the most frequent contributing cause and age-related macular degeneration the second most frequent contributing cause of visual impairment. The overall prevalence proportions of age-related macular degeneration, cataract, diabetic retinopathy, and definite glaucoma were 33.6%, 39.6%, 20.1%, and 5.6%, respectively. Most of the other eye abnormalities were rare in the study participants, except for pinguecula (42.4%) and dermatochalasis (30.1%). CONCLUSIONS: Oklahoma Indians have a higher prevalence of visual impairment, age-related macular degeneration, and diabetic retinopathy than other ethnic groups. The implementation of adequate treatment and prevention programs for eye diseases is indicated. PMID- 16344443 TI - Microsatellite instability and alterations of mismatch repair protein expression in choroidal melanomas. AB - OBJECTIVES: To examine choroidal melanomas for genomic instability, manifested by microsatellite instability (MSI) and mismatch repair (MMR) protein alterations, and to determine the association of these alterations with selected clinicopathological features of the tumors. METHODS: Polymerase chain reaction based microsatellite assays were applied to analyze 57 cases of choroidal melanomas using 11 microsatellite markers at 5 chromosomal regions: 1p, 2p, 4q, 9p, and 17p. Immunoperoxidase staining methods and mouse monoclonal antibodies were used to investigate the expression patterns of MMR proteins. RESULTS: Microsatellite instability was found at the 1p, 9p, and 17p regions in these lesions with an overall prevalence of 35% (20/57). The frequency of MSI ranged from 9% (1/11) to 27% (3/11), ie, low-level MSI (MSI-L). The instability was most commonly found at the 1p region (D1S2734, 55%; D1S2832, 40%; and D1S233, 20%). Two MSI banding patterns, band shifts and the appearance of additional bands, were found in 10% and 90% of the unstable lesions, respectively. The average percentages of hMLH1 and hMSH2 positively stained cells were insignificantly reduced in the unstable lesions (81.7 +/- 9.3 and 76.7 +/- 16.7) as compared with the stable lesions (84.1 +/- 15.5 and 78.6 +/- 19.6; P = .62 and 0.74 for hMLH1 and hMSH2, respectively). There was no significant difference in survival between the 2 groups; however, relative to the stable subset, the unstable tumors showed a trend (P<.10) toward occurring at a younger age and having tumor cells in vascular lakes. CONCLUSIONS: The presence of MSI-L in some choroidal melanomas defines a novel genetic subset of these tumors and suggests that MSI (genomic instability) may play a role in their molecular pathogenesis. Elucidation of the underlying mechanisms for MSI will require further investigation. CLINICAL RELEVANCE: Detection of the MSI-L pattern might prove to be useful as an adjunct to the conventional diagnosis of choroidal melanomas. Larger series are needed to determine whether any of the correlative trends noted in this study will achieve statistical significance. To the best of our knowledge, this study is the first to define both the MSI and MMR protein expression features of choroidal melanomas. PMID- 16344444 TI - Spectral domain optical coherence tomography: ultra-high speed, ultra-high resolution ophthalmic imaging. AB - OBJECTIVE: To introduce a new ophthalmic optical coherence tomography technology that allows unprecedented simultaneous ultra-high speed and ultra-high resolution. METHODS: Using a superluminescent diode source, a clinically viable ultra-high speed, ultra-high resolution spectral domain optical coherence tomography system was developed. RESULTS: In vivo images of the retina, the optic nerve head, and retinal blood flow were obtained at an ultra-high speed of 34.1 microseconds (ms) per A-scan, which is 73 times faster than commercially available optical coherence tomography instruments. Single images (B-scans) consisting of 1000 A-scans were acquired in 34.1 ms, allowing video rate imaging at 29 frames per second with an axial resolution of 6 mum. Using a different source in a slightly slower configuration, single images consisting of 500 A scans were acquired in 34 ms, allowing imaging at 29 frames per second at an axial resolution of 3.5 microm, which is 3 times better than commercially available optical coherence tomography instruments. The amount of energy directed into the eye in both cases, 600 microW, is less than that of the Stratus OCT3 and is safe for intrabeam viewing for up to 8 hours at the same retinal location. CONCLUSION: Spectral domain optical coherence tomography technology enables ophthalmic imaging with unprecedented simultaneous ultra-high speed and ultra high resolution. PMID- 16344445 TI - Intracorneal injection of amphothericin B for recurrent fungal keratitis and endophthalmitis. AB - Penetrating keratoplasty carries an infectious risk. Its requirement for topical corticosteroid therapy facilitates fungal growth with resulting keratitis. Although progression of fungal keratitis to intraocular infection is uncommon, endophthalmitis resulting from keratitis usually has a poor visual prognosis. Fungal infection under these circumstances remains a diagnostic and therapeutic challenge. We report a complicated case of recurrent fungal keratitis with endophthalmitis following a contaminated penetrating keratoplasty that ultimately was controlled with a new treatment modality. Intrastromal corneal injections combined with intravitreal injection of amphotericin B led to the eradication of the corneal fungal plaques and the intraocular infection. Intrastromal corneal injections of amphotericin B may offer a less invasive, in-office alternative to repeat penetrating keratoplasty. PMID- 16344446 TI - Tissue bioengineering: potential applications to glaucoma. PMID- 16344447 TI - Effect of physician reimbursement methodology on the rate and cost of cataract surgery. AB - OBJECTIVES: To compare the effects of 2 reimbursement methodologies, fee-for service and contact capitation, on cataract extraction rates and costs in a stable physician population with little potential for the influence of patient selection. Previous research evaluating the relationship between physician reimbursement incentives and cataract surgical rates has been limited by physician and patient selection bias. METHODS: A pre-post analysis of claims and encounter data for an average of 91,473 commercial beneficiaries and 14,084 Medicare beneficiaries receiving eye care from a network of ophthalmologists and optometrists in St Louis, Mo, between 1997 and 1998. The rate of cataract extractions per 1000 beneficiaries, the costs of cataract procedures, the rates of noncataract procedures, and the level of professional reimbursement for providers were compared during the final 6 months of fee-for-service physician reimbursement and the first 6 months of contact capitation. RESULTS: Both commercial and Medicare beneficiaries were approximately one half as likely to have cataract extraction (P<.001) under contact capitation as compared with fee for-service. Professional reimbursement increased by 8% whereas facility fees for cataract procedures decreased by approximately 45%. Cataract surgical rates were disproportionately affected when compared with other ophthalmologic procedures. During the study period, cataract surgical rates were stable in the national and Missouri traditional fee-for-service Medicare population. CONCLUSIONS: The stability of the physician and patient populations allowed us to isolate the effects of physician reimbursement methodology on practice patterns. Compared with fee-for-service, contact capitation reimbursement was associated with significant decreases in cataract extraction rates and costs. The frequency of the cataract extraction surgery, the most common major elective procedure in ophthalmology, was more responsive to physician financial incentives than other ophthalmologic procedures were. PMID- 16344448 TI - Elegant diagnosis. PMID- 16344449 TI - Retinal anastomosis to choroidal neovascularization: a bum rap for a difficult disease. PMID- 16344450 TI - Strong labeling for iron and the iron-handling proteins ferritin and ferroportin in the photoreceptor layer in age-related macular degeneration. PMID- 16344451 TI - Optical coherence tomographic findings of combined hamartoma of the retina and retinal pigment epithelium in 11 patients. PMID- 16344452 TI - Dynamic atypical optic nerve coloboma associated with transient macular detachment. PMID- 16344453 TI - Extensive chorioretinitis and severe vision loss associated with west nile virus meningoencephalitis. PMID- 16344454 TI - Retinal folds as initial manifestation of orbital lymphangioma. PMID- 16344455 TI - Nocardia keratitis in a contact lens wearer. PMID- 16344456 TI - Convergence insufficiency: randomized clinical trial. PMID- 16344457 TI - When the B-team runs plasticity: GluR2 receptor trafficking in cerebellar long term potentiation. PMID- 16344458 TI - In vivo targeting of dendritic cells for activation of cellular immunity using vaccine carriers based on pH-responsive microparticles. AB - Activating the immune system to trigger a specific response is a major challenge in vaccine development. In particular, activating sufficient cytotoxic T lymphocyte-mediated cellular immunity, which is crucial for the treatment of many diseases including cancer and AIDS, has proven to be especially challenging. In this study, antigens were encapsulated in acid-degradable polymeric particle carriers to cascade cytotoxic T lymphocyte activation. To target dendritic cells, the most potent antigen-presenting cells, the particle carriers, were further conjugated with monoclonal antibodies. A series of ex vivo and in vivo studies have shown increased receptor-mediated uptake of antibody-conjugated particles by dendritic cells as well as migration of particle-carrying dendritic cells to lymph nodes and stimulation of naive T cells leading to enhanced cellular immune response as confirmed by specific cell lysis and IFN-gamma secretion. PMID- 16344459 TI - The zebrafish kohtalo/trap230 gene is required for the development of the brain, neural crest, and pronephric kidney. AB - Mutation of the gene encoding the Mediator component thyroid hormone receptor associated protein (TRAP)230/MED12 affects the development of multiple systems in zebrafish embryogenesis. We isolated two ethylnitrosourea-induced alleles in the gene encoding this protein and named the locus kohtalo (kto) after the homologous locus in Drosophila. Homozygous kto mutant zebrafish embryos show defects in brain, neural crest, and kidney development and die at approximately 6 days postfertilization. In the affected tissues, differentiation is initiated and many cell type-specific genes are expressed, but there is a failure of morphogenesis and failure to complete differentiation. These results suggest that critical targets of TRAP230 function may include proteins important for cell mobility, cell sorting, and tissue assembly. PMID- 16344460 TI - The chemistry of child neglect: do oxytocin and vasopressin mediate the effects of early experience? PMID- 16344461 TI - Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. AB - In a recent report, [Zhang et al. (2003) N. Engl. J. Med. 348, 203-213], the presence of CD3+ tumor-infiltrating lymphocytes (TILs) was found to correlate with improved survival in epithelial ovarian cancer. We performed immunohistochemical analysis for TILs and cancer testis antigens in 117 cases of epithelial ovarian cancer. The interrelationship between subpopulations of TILs and expression of cancer testis antigens was investigated, as well as between TILs and overall survival. The median follow-up of the patients was 31 months. Patients with higher frequencies of intraepithelial CD8+ T cells demonstrated improved survival compared with patients with lower frequencies [median = 55 versus 26 months; hazard ratio = 0.33; confidence interval (C.I.) = 0.18-0.60; P = 0.0003]. No association was found for CD3+ TILs or other subtypes of intraepithelial or stromal TILs. However, the subgroups with high versus low intraepithelial CD8+/CD4+ TIL ratios had median survival of 74 and 25 months, respectively (hazard ratio = 0.30; C.I. = 0.16-0.55; P = 0.0001). These results indicate that CD4+ TILs influence the beneficial effects of CD8+ TIL. This unfavorable effect of CD4+ T cells on prognosis was found to be due to CD25+ forkhead box P3 (FOXP3)+ regulatory T cells (Treg; suppressor T cells), as indicated by survival of patients with high versus low CD8+/Treg ratios (median = 58 versus 23 months; hazard ratio = 0.31; C.I. = 0.17-0.58; P = 0.0002). The favorable prognostic effect of intraepithelial CD8+ TILs did not correlate with concurrent expression of NY-ESO-1 or MAGE antigens. We conclude that intraepithelial CD8+ TILs and a high CD8+/Treg ratio are associated with favorable prognosis in epithelial ovarian cancer. PMID- 16344462 TI - Differential cis-regulation of human versus mouse TERT gene expression in vivo: identification of a human-specific repressive element. AB - In vivo expression of human telomerase is significantly different from that of mouse telomerase. To assess the basis for this difference, a bacterial artificial chromosome clone containing the entire hTERT (human telomerase reverse transcriptase) gene was introduced in mice. In these transgenic mice, expression of the hTERT transgene was similar to that of endogenous hTERT in humans, rather than endogenous mTERT (mouse telomerase reverse transcriptase). In tissues and cells showing a striking difference in expression levels between hTERT in humans and mTERT in mice (i.e., liver, kidney, lung, uterus, and fibroblasts), expression of the hTERT transgene in transgenic mice was repressed, mimicking hTERT in humans. The transcriptional activity of the hTERT promoter was much lower than that of the mTERT promoter in mouse embryonic fibroblasts or human fibroblasts. Mutational analysis of the hTERT and mTERT promoters revealed that a nonconserved GC-box within the hTERT promoter was responsible for the human specific repression. These results reveal that a difference in cis-regulation of transcription, rather than transacting transcription factors, is critical to species differences in tissue-specific TERT expression. Our data also suggest that the GC-box-mediated, human-specific mechanism for TERT repression is impaired in human cancers. This study represents a detailed characterization of the functional difference in a gene promoter of mice versus humans and provides not only important insight into species-specific regulation of telomerase and telomeres but also an experimental basis for generating mice humanized for telomerase enzyme and its pattern of expression. PMID- 16344464 TI - Cranial morphology of early Americans from Lagoa Santa, Brazil: implications for the settlement of the New World. AB - Comparative morphological studies of the earliest human skeletons of the New World have shown that, whereas late prehistoric, recent, and present Native Americans tend to exhibit a cranial morphology similar to late and modern Northern Asians (short and wide neurocrania; high, orthognatic and broad faces; and relatively high and narrow orbits and noses), the earliest South Americans tend to be more similar to present Australians, Melanesians, and Sub-Saharan Africans (narrow and long neurocrania; prognatic, low faces; and relatively low and broad orbits and noses). However, most of the previous studies of early American human remains were based on small cranial samples. Herein we compare the largest sample of early American skulls ever studied (81 skulls of the Lagoa Santa region) with worldwide data sets representing global morphological variation in humans, through three different multivariate analyses. The results obtained from all multivariate analyses confirm a close morphological affinity between South-American Paleoindians and extant Australo-Melanesians groups, supporting the hypothesis that two distinct biological populations could have colonized the New World in the Pleistocene/Holocene transition. PMID- 16344463 TI - Preferential occupancy of histone variant H2AZ at inactive promoters influences local histone modifications and chromatin remodeling. AB - The yeast histone variant H2AZ (Htz1) is implicated in transcription activation, prevention of the ectopic spread of heterochromatin, and genome integrity. Our genome-wide localization analysis revealed that Htz1 is widely, but nonrandomly, distributed throughout the genome in an SWR1-dependent manner. We found that Htz1 is enriched in intergenic regions compared with coding regions. Its occupancy is inversely proportional to transcription rates and the enrichment of the RNA polymerase II under different growth conditions. However, Htz1 does not seem to directly regulate transcription repression genome-wide; instead, the presence of Htz1 under the inactivated condition is essential for optimal activation of a subset of genes. In addition, Htz1 is not generally responsible for nucleosome positioning, even at those promoters where Htz1 is highly enriched. Finally, using a biochemical approach, we demonstrate that incorporation of Htz1 into nucleosomes inhibits activities of histone modifiers associated with transcription, Dot1, Set2, and NuA4 and reduces the nucleosome mobilization driven by chromatin remodeling complexes. These lines of evidence collectively suggest that Htz1 may serve to mark quiescent promoters for proper activation. PMID- 16344465 TI - Introduction of an additional pathway for lactate oxidation in the treatment of lactic acidosis and mitochondrial dysfunction in Caenorhabditis elegans. AB - Mitochondrial dysfunction, with an estimated incidence of 1 in 5,000 births, is associated with a wide variety of multisystem degenerative diseases. Among the most prevalent forms of dysfunction are defects in the NADH:ubiquinone oxidoreductase (complex I). Caenorhabditis elegans strains with complex I mutations exhibit characteristic features of human mitochondrial disease including decreased rates of respiration and lactic acidosis. We hypothesized that introducing an additional pathway for the direct oxidation of lactate would be beneficial for energy metabolism. The yeast CYB2 gene encodes an L lactate:cytochrome c oxidoreductase that oxidizes lactate, donates electrons directly into the mitochondrial respiratory chain, and supports lactate-dependent respiration. Cyb2p expression markedly increases lifespan, fertility, respiration rates, and ATP content in complex I-deficient animals. Our results indicate that metabolic imbalance leading to lactic acidosis and energy depletion are central mechanisms of pathogenesis in mitochondrial dysfunction and that introduction of an additional pathway for lactate oxidation should be considered as a treatment. PMID- 16344466 TI - Human La is found at RNA polymerase III-transcribed genes in vivo. AB - The human La autoantigen can bind to nascent RNA transcripts and has also been postulated to act as an RNA polymerase III (pol III) transcription initiation and termination factor. Here, we show by chromatin immunoprecipitation (ChIP) that La is associated with pol III-transcribed genes in vivo. In contrast, the Ro autoantigen, which can also bind pol III transcripts, is not found at these genes. The putative pol III transcription factors NF1 and TFIIA are also not detected at class III genes. Binding of La remains when transcription is repressed at mitosis and does not correlate with the presence of polymerase at the gene. However, gene occupancy depends on the phosphorylation status of La, with the less prevalent, unphosphorylated form being found selectively on pol III templates. PMID- 16344467 TI - Crystal structure of receptor-binding C-terminal repeats from Clostridium difficile toxin A. AB - Clostridium difficile is a major nosocomial pathogen that produces two large protein toxins [toxin A (TcdA) and toxin B (TcdB)] capable of disrupting intestinal epithelial cells. Both belong to the family of large clostridial cytotoxins, which are characterized by the presence of a repetitive C-terminal repetitive domain (CRD). In TcdA, the CRD is composed of 39 repeats that are responsible for binding to cell surface carbohydrates. To understand the molecular structural basis of cell binding by the toxins from C. difficile, we have determined a 1.85-A resolution crystal structure of a 127-aa fragment from the C terminus of the toxin A CRD. This structure reveals a beta-solenoid fold containing five repeats, with each repeat consisting of a beta-hairpin followed by a loop of 7-10 residues in short repeats (SRs) or 18 residues in long repeats (LRs). Adjacent pairs of beta-hairpins are related to each other by either 90 degree or 120 degree screw-axis rotational relationships, depending on the nature of the amino acids at key positions in adjacent beta-hairpins. Models of the complete CRDs of toxins A and B suggest that each CRD contains straight stretches of beta-solenoid composed of three to five SRs that are punctuated by kinks introduced by the presence of a single LR. These structural features provide a framework for understanding how large clostridial cytotoxins bind to cell surfaces and suggest approaches for developing novel treatments for C. difficile associated diseases by blocking the binding of toxins to cell surfaces. PMID- 16344468 TI - Ubiquitinated proliferating cell nuclear antigen activates translesion DNA polymerases eta and REV1. AB - In response to DNA damage, the Rad6/Rad18 ubiquitin-conjugating complex monoubiquitinates the replication clamp proliferating cell nuclear antigen (PCNA) at Lys-164. Although ubiquitination of PCNA is recognized as an essential step in initiating postreplication repair, the mechanistic relevance of this modification has remained elusive. Here, we describe a robust in vitro system that ubiquitinates yeast PCNA specifically on Lys-164. Significantly, only those PCNA clamps that are appropriately loaded around effector DNA by its loader, replication factor C, are ubiquitinated. This observation suggests that, in vitro, only PCNA present at stalled replication forks is ubiquitinated. Ubiquitinated PCNA displays the same replicative functions as unmodified PCNA. These functions include loading onto DNA by replication factor C, as well as Okazaki fragment synthesis and maturation by the PCNA-coordinated actions of DNA polymerase delta, the flap endonuclease FEN1, and DNA ligase I. However, whereas the activity of DNA polymerase zeta remains unaffected by ubiquitination of PCNA, ubiquitinated PCNA specifically activates two key enzymes in translesion synthesis: DNA polymerase eta, the yeast Xeroderma pigmentosum ortholog, and Rev1, a deoxycytidyl transferase that functions in organizing the mutagenic DNA replication machinery. We propose that ubiquitination of PCNA increases its functionality as a sliding clamp to promote mutagenic DNA replication. PMID- 16344469 TI - The crystal structure of CREG, a secreted glycoprotein involved in cellular growth and differentiation. AB - The cellular repressor of E1A-stimulated genes (CREG) is a secreted glycoprotein that inhibits proliferation and enhances differentiation of human embryonal carcinoma cells. CREG binds to the cation-independent mannose 6-phosphate (M6P)/insulin-like growth factor II (IGF2) receptor (IGF2R) (M6P/IGF2R), and this receptor has been shown to be required for CREG-induced growth suppression. To better understand CREG function in cellular growth and differentiation, we solved the 3D crystal structure of this protein to 1.9-A resolution. CREG forms a tight homodimeric complex, and CREG monomers display a beta-barrel fold. The three potential glycosylation sites on CREG map to a confined patch opposite the dimer interface. Thus, dimerization of glycosylated CREG likely presents a bivalent ligand for the M6P/IGF2R. Closely related structural homologs of CREG are FMN binding split-barrel fold proteins that bind flavin mononucleotide. Our structure shows that the putative flavin mononucleotide-binding pocket in CREG is sterically blocked by a loop and several key bulky residues. A mutant of CREG lacking a part of this loop maintained overall structure and dimerization, as well as M6P/IGF2R binding, but lost the growth suppression activity of WT CREG. Thus, analysis of a structure-based mutant of CREG revealed that binding to M6P/IGF2R, while necessary, is not sufficient for CREG-induced growth suppression. These findings indicate that CREG utilizes a known fold for a previously undescribed function [corrected] PMID- 16344470 TI - Volumetric tomography of fluorescent proteins through small animals in vivo. AB - Volumetric detection and accurate quantification of fluorescent proteins in entire animals would greatly enhance our ability to monitor biological processes in vivo. Here we present a quantitative tomographic technique for visualization of superficial and deep-seated (>2-3 mm) fluorescent protein activity in vivo. We demonstrate noninvasive imaging of lung tumor progression in a murine model, as well as imaging of gene delivery using a herpes virus vector. This technology can significantly improve imaging capacity over the current state of the art and should find wide in vivo imaging applications in drug discovery, immunology, and cancer research. PMID- 16344471 TI - Notch deficiency implicated in the pathogenesis of congenital disorder of glycosylation IIc. AB - Congenital disorder of glycosylation IIc (CDG IIc), also termed leukocyte adhesion deficiency II, is a recessive syndrome characterized by slowed growth, mental retardation, and severe immunodeficiency. Recently, the gene responsible for CDG IIc was found to encode a GDP-fucose transporter. Here, we investigated the possible cause of the developmental defects in CDG IIc patients by using a Drosophila model. Biochemically, we demonstrated that a Drosophila homolog of the GDP-fucose transporter, the Golgi GDP-fucose transporter (Gfr), specifically transports GDP-fucose in vitro. To understand the function of the Gfr gene, we generated null mutants of Gfr in Drosophila. The phenotypes of the Drosophila Gfr mutants were rescued by the human GDP-fucose transporter transgene. Our phenotype analyses revealed that Notch (N) signaling was deficient in these Gfr mutants. GDP-fucose is known to be essential for the fucosylation of N-linked glycans and for O-fucosylation, and both fucose modifications are present on N. Our results suggest that Gfr is involved in the fucosylation of N-linked glycans on N and its O-fucosylation, as well as those of bulk proteins. However, despite the essential role of N O-fucosylation during development, the Gfr homozygote was viable. Thus, our results also indicate that the Drosophila genome encodes at least another GDP fucose transporter that is involved in the O-fucosylation of N. Finally, we found that mammalian Gfr is required for N signaling in mammalian cultured cells. Therefore, our results implicate reduced N signaling in the pathology of CDG IIc. PMID- 16344472 TI - Up-regulation of beta-catenin by a viral oncogene correlates with inhibition of the seven in absentia homolog 1 in B lymphoma cells. AB - The protein levels of beta-catenin are tightly regulated by the ubiquitin/proteasome system. We provide evidence that two distinct ubiquitin dependent degradation pathways for beta-catenin are active in the same Burkitt's lymphoma cells: Along with the classical glycogen-synthase kinase 3beta-dependent destruction machinery, degradation of beta-catenin through seven in absentia homolog 1 (Siah-1) ubiquitin ligase is functional in these cells. We show that inhibition of endogenous Siah-1 stabilizes and activates beta-catenin in B cells. The principal Epstein-Barr virus oncoprotein, latent membrane protein 1, is involved in beta-catenin up-regulation, and expression of latent membrane protein 1 in B lymphoma cells is associated with decreased Siah-1 RNA and protein levels. Thus, we demonstrate the significance of the endogenous Siah-1-dependent ubiquitin/proteasome pathway for beta-catenin degradation in malignant human cells and its regulation by a viral oncogene. PMID- 16344474 TI - Hyperpolarized krypton-83 as a contrast agent for magnetic resonance imaging. AB - For the first time, magnetic resonance imaging (MRI) with hyperpolarized (hp) krypton-83 (83Kr) has become available. The relaxation of the nuclear spin of 83Kr atoms (I = 9/2) is driven by quadrupolar interactions during brief adsorption periods on surrounding material interfaces. Experiments in model systems reveal that the longitudinal relaxation of hp 83Kr gas strongly depends on the chemical composition of the materials. The relaxation-weighted contrast in hp 83Kr MRI allows for the distinction between hydrophobic and hydrophilic surfaces. The feasibility of hp 83Kr MRI of airways is tested in canine lung tissue by using krypton gas with natural abundance isotopic distribution. Additionally, the influence of magnetic field strength and the presence of a breathable concentration of molecular oxygen on longitudinal relaxation are investigated. PMID- 16344475 TI - Unexceptional sharpness of frequency tuning in the human cochlea. AB - The responses to sound of auditory-nerve fibers are well known in many animals but are topics of conjecture for humans. Some investigators have claimed that the auditory-nerve fibers of humans are more sharply tuned than are those of various experimental animals. Here we invalidate such claims. First, we show that forward masking psychophysical tuning curves, which were used as the principal support for those claims, greatly overestimate the sharpness of cochlear tuning in experimental animals and, hence, also probably in humans. Second, we calibrate compound action potential tuning curves against the tuning of auditory-nerve fibers in experimental animals and use compound action potential tuning curves recorded in humans to show that the sharpness of tuning in human cochleae is not exceptional and that it is actually similar to tuning in all mammals and birds for which comparisons are possible. Third, we note that the similarity of frequency of tuning across species with widely diverse cochlear lengths and auditory bandwidths implies that for any given stimulus frequency the "cochlear amplifier" is confined to a highly localized region of the cochlea. PMID- 16344476 TI - Three independent biological mechanisms cause exercise-associated hyponatremia: evidence from 2,135 weighed competitive athletic performances. AB - To evaluate the role of fluid and Na+ balance in the development of exercise associated hyponatremia (EAH), changes in serum Na+ concentrations ([Na+]) and in body weight were analyzed in 2,135 athletes in endurance events. Eighty-nine percent of athletes completed these events either euhydrated (39%) or with weight loss (50%) and with normal (80%) or elevated (13%) serum [Na+]. Of 231 (11%) athletes who gained weight during exercise, 70% were normonatremic or hypernatremic, 19% had a serum [Na+] between 129-135 mmol/liter, and 11% a serum [Na+] of <129 mmol/liter. Serum [Na+] after racing was a linear function with a negative slope of the body weight change during exercise. The final serum [Na+] in a subset of 18 subjects was predicted from the amount of Na+ that remained osmotically inactive at the completion of the trial. Weight gain consequent to excessive fluid consumption was the principal cause of a reduced serum [Na+] after exercise, yet most (70%) subjects who gained weight maintained or increased serum [Na+], requiring the addition of significant amounts of Na+ (>500 mmol) into an expanded volume of total body water. This Na+ likely originated from osmotically inactive, exchangeable stores. Thus, EAH occurs in athletes who (i) drink to excess during exercise, (ii) retain excess fluid because of inadequate suppression of antidiuretic hormone secretion, and (iii) osmotically inactivate circulating Na+ or fail to mobilize osmotically inactive sodium from internal stores. EAH can be prevented by insuring that athletes do not drink to excess during exercise, which has been known since 1985. PMID- 16344477 TI - Snapshots of cytochrome c folding. AB - Dansyl-to-heme distance distributions [P(r)] during folding have been determined in five variants of Saccharomyces cerevisiae iso-1 ferricytochrome c (labeled at mutant Cys residues 4, 39, 50, 66, and 99) by analysis of fluorescence energy transfer kinetics. Moment analysis of the P(r) distributions clearly indicates that cytochrome c refolding is not a simple two-state process. After 1 ms of folding, the polypeptide ensemble is not uniformly collapsed and there are site variations in the relative populations of collapsed structures. P(r) distributions reveal structural features of the multiple intermediate species and evolution of the polypeptide ensemble. PMID- 16344473 TI - The planarian Schmidtea mediterranea as a model for epigenetic germ cell specification: analysis of ESTs from the hermaphroditic strain. AB - Freshwater planarians have prodigious regenerative abilities that enable them to form complete organisms from tiny body fragments. This plasticity is also exhibited by the planarian germ cell lineage. Unlike many model organisms in which germ cells are specified by localized determinants, planarian germ cells appear to be specified epigenetically, arising postembryonically from stem cells. The planarian Schmidtea mediterranea is well suited for investigating the mechanisms underlying epigenetic germ cell specification. Two strains of S. mediterranea exist: a hermaphroditic strain that reproduces sexually and an asexual strain that reproduces by means of transverse fission. To date, expressed sequence tags (ESTs) have been generated only from the asexual strain. To develop molecular reagents for studying epigenetic germ cell specification, we have sequenced 27,161 ESTs from two developmental stages of the hermaphroditic strain of S. mediterranea; this collection of ESTs represents approximately 10,000 unique transcripts. blast analysis of the assembled ESTs showed that 66% share similarity to sequences in public databases. We annotated the assembled ESTs using Gene Ontology terms as well as conserved protein domains and organized them in a relational database. To validate experimentally the Gene Ontology annotations, we used whole-mount in situ hybridization to examine the expression patterns of transcripts assigned to the biological process "reproduction." Of the 53 genes in this category, 87% were expressed in the reproductive organs. In addition to its utility for studying germ cell development, this EST collection will be an important resource for annotating the planarian genome and studying this animal's amazing regenerative abilities. PMID- 16344478 TI - Apolipoprotein (apo) E4 enhances amyloid beta peptide production in cultured neuronal cells: apoE structure as a potential therapeutic target. AB - Apolipoprotein (apo) E4 is a major risk factor for Alzheimer's disease, and many studies have suggested that apoE has isoform-specific effects on the deposition or clearance of amyloid beta (Abeta) peptides. We examined the effects of apoE isoforms on the processing of amyloid precursor protein (APP) and on Abeta production in rat neuroblastoma B103 cells stably transfected with human wild type APP695 (B103-APP). Lipid-poor apoE4 increased Abeta production in B103-APP cells to a greater extent than lipid-poor apoE3 (60% vs. 30%) due to more pronounced stimulation of APP recycling by apoE4 than apoE3. The difference in Abeta production was abolished by preincubating the cells with the receptor associated protein (25 nM), which blocks the low-density lipoprotein receptor related protein (LRP) pathway, or by reducing LRP expression by small interference RNA. The differences were also attenuated by replacing Arg-61 with threonine in apoE4 or pretreating apoE4 with small molecules, both of which abolish apoE4 intramolecular domain interaction. Thus, apoE4 appears to modulate APP processing and Abeta production through both the LRP pathway and domain interaction. These findings provide insights into why apoE4 is associated with increased risk for Alzheimer's disease and may represent a potential target for drug development. PMID- 16344479 TI - Lipid- and receptor-binding regions of apolipoprotein E4 fragments act in concert to cause mitochondrial dysfunction and neurotoxicity. AB - Apolipoprotein (apo) E4, a 299-aa protein and a major risk factor for Alzheimer's disease, can be cleaved to generate C-terminal-truncated fragments that cause neurotoxicity in vitro and neurodegeneration and behavioral deficits in transgenic mice. To investigate this neurotoxicity, we expressed apoE4 with C- or N-terminal truncations or mutations in transfected Neuro-2a cells. ApoE4 (1-272) was neurotoxic, but full-length apoE4(1-299) and apoE4(1-240) were not, suggesting that the lipid-binding region (amino acids 241-272) mediates the neurotoxicity and that amino acids 273-299 are protective. A quadruple mutation in the lipid-binding region (I250A, F257A, W264R, and V269A) abolished the neurotoxicity of apoE4(1-272), and single mutations in the region of amino acids 273-299 (L279Q, K282A, or Q284A) made full-length apoE4 neurotoxic. Immunofluorescence staining showed that apoE4(1-272) formed filamentous inclusions containing phosphorylated tau in some cells and interacted with mitochondria in others, leading to mitochondrial dysfunction as determined by MitoTracker staining and flow cytometry. ApoE4(241-272) did not cause mitochondrial dysfunction or neurotoxicity, suggesting that the lipid-binding region alone is insufficient for neurotoxicity. Truncation of N-terminal sequences (amino acids 1-170) containing the receptor-binding region (amino acids 135-150) and triple mutations within that region (R142A, K146A, and R147A) abolished the mitochondrial interaction and neurotoxicity of apoE4(1-272). Further analysis showed that the receptor-binding region is required for escape from the secretory pathway and that the lipid-binding region mediates mitochondrial interaction. Thus, the lipid- and receptor-binding regions in apoE4 fragments act together to cause mitochondrial dysfunction and neurotoxicity, which may be important in Alzheimer's disease pathogenesis. PMID- 16344480 TI - The cell-shape protein MreC interacts with extracytoplasmic proteins including cell wall assembly complexes in Caulobacter crescentus. AB - The bacterial actin homolog, MreB, forms helical cables within the cell that are required for maintenance of a rod shape. These helical structures are thought to be involved in the spatial organization of cell wall (peptidoglycan) synthesizing complexes of penicillin-binding proteins (PBPs). Here, we examined the role of the MreC cell shape protein in this process in Caulobacter crescentus. Subcellular fractionation experiments showed that MreC is a periplasmic protein and, as assayed by immunofluorescence microscopy, adopted helical or banded patterns along the cell length reminiscent of those formed by MreB and PBP2. The pattern of MreC and PBP2 localization remained when MreB cables were disrupted by treatment with the inhibitor A22. However, long-term absence of MreB led to cell shape changes and an eventual loss of MreC localization, suggesting that an independent structure, perhaps an intact peptidoglycan layer, contributes to the MreC localization pattern. Using affinity chromatography with MreC covalently bound to Sepharose, we isolated several PBPs from cell extracts that eluted from the column as heterogeneous complexes. In this same experiment, using mass spectrometry-based protein identification, we identified several outer membrane proteins, including TonB-dependent receptor transport proteins, that interacted with MreC. Imaging live cells containing fusions of these outer membrane proteins to green fluorescent protein showed that they adopted a subcellular localization pattern that was similar to that of MreC. These results suggest that MreC may function in the spatial organization of PBPs as well as other proteins that lie outside the cytoplasmic membrane. PMID- 16344481 TI - Two independent spiral structures control cell shape in Caulobacter. AB - The actin homolog MreB contributes to bacterial cell shape. Here, we explore the role of the coexpressed MreC protein in Caulobacter and show that it forms a periplasmic spiral that is out of phase with the cytoplasmic MreB spiral. Both mreB and mreC are essential, and depletion of either protein results in a similar cell shape defect. MreB forms dynamic spirals in MreC-depleted cells, and MreC localizes helically in the presence of the MreB-inhibitor A22, indicating that each protein can form a spiral independently of the other. We show that the peptidoglycan transpeptidase Pbp2 also forms a helical pattern that partially colocalizes with MreC but not MreB. Perturbing either MreB (with A22) or MreC (with depletion) causes GFP-Pbp2 to mislocalize to the division plane, indicating that each is necessary but not sufficient to generate a helical Pbp2 pattern. We show that it is the division process that draws Pbp2 to midcell in the absence of MreB's regulation, because cells depleted of the tubulin homolog FtsZ maintain a helical Pbp2 localization in the presence of A22. By developing and employing a previously uncharacterized computational method for quantitating shape variance, we find that a FtsZ depletion can also partially rescue the A22-induced shape deformation. We conclude that MreB and MreC form spatially distinct and independently localized spirals and propose that MreB inhibits division plane localization of Pbp2, whereas MreC promotes lengthwise localization of Pbp2; together these two mechanism ensure a helical localization of Pbp2 and, thereby, the maintenance of proper cell morphology in Caulobacter. PMID- 16344482 TI - Targeted ablation of plectin isoform 1 uncovers role of cytolinker proteins in leukocyte recruitment. AB - Plectin, a typical cytolinker protein, is essential for skin and skeletal muscle integrity. It stabilizes cells mechanically, regulates cytoskeleton dynamics, and serves as a scaffolding platform for signaling molecules. A variety of isoforms expressed in different tissues and cell types account for this versatility. To uncover the role of plectin 1, the major isoform expressed in tissues of mesenchymal origin, against the background of all other variants, we raised plectin isoform 1-specific antibodies and generated isoform-deficient mice. In contrast to plectin-null mice (lacking all plectin isoforms), which die shortly after birth because of severe skin blistering, plectin isoform 1-deficient mice were viable at birth, had a normal lifespan, and did not display the skin blistering phenotype. However, dermal fibroblasts isolated from plectin 1 deficient mice exhibited abnormalities in their actin cytoskeleton and impaired migration potential. Similarly, plectin 1-deficient T cells isolated from nymph nodes showed diminished chemotactic migration in vitro. Most strikingly, in vivo we found that leukocyte infiltration during wound healing was reduced in the mutant mice. These data show a specific role of a cytolinker protein in immune cell motility. Single isoform-deficient mice thus represent a powerful tool to unravel highly specific functions of plectin variants. PMID- 16344483 TI - Structure of the streptococcal cell wall C5a peptidase. AB - The structure of a cell surface enzyme from a gram-positive pathogen has been determined to 2-A resolution. Gram-positive pathogens have a thick cell wall to which proteins and carbohydrate are covalently attached. Streptococcal C5a peptidase (SCP), is a highly specific protease and adhesin/invasin. Structural analysis of a 949-residue fragment of the [D130A,S512A] mutant of SCP from group B Streptococcus (S. agalactiae, SCPB) revealed SCPB is composed of five distinct domains. The N-terminal subtilisin-like protease domain has a 134-residue protease-associated domain inserted into a loop between two beta-strands. This domain also contains one of two Arg-Gly-Asp (RGD) sequences found in SCPB. At the C terminus are three fibronectin type III (Fn) domains. The second RGD sequence is located between Fn1 and Fn2. Our analysis suggests that SCP binding to integrins by the RGD motifs may stabilize conformational changes required for substrate binding. PMID- 16344484 TI - The proficiency of a thermophilic chorismate mutase enzyme is solely through an entropic advantage in the enzyme reaction. AB - A study of the Thermus thermophilus chorismate mutase (TtCM) is described by using quantum mechanics (self-consistent-charge density-functional tight binding)/molecular mechanics, umbrella sampling, and the weighted histogram analysis method. The computed free energies of activation for the reactions in water and TtCM are comparable to the experimental values. The free energies for formation of near attack conformer have been determined to be 8.06 and 0.05 kcal/mol in water and TtCM, respectively. The near attack conformer stabilization contributes approximately 90% to the proficiency of the enzymatic reaction compared with the reaction in water. The transition state (TS) structures and partial atom charges are much the same in the enzymatic and water reactions. The difference in the electrostatic interactions of Arg-89 with O13 in the enzyme substrate complex and enzyme-TS complex provides the latter with but 0.55 kcal/mol of 1.92 kcal/mol total TS stabilization. Differences in electrostatic interactions between components at the active site in the enzyme-substrate complex and enzyme-TS complex are barely significant, such that TS stabilization is of minor importance and the enzymatic catalysis is through an entropic advantage. PMID- 16344485 TI - Pinpointing and preventing imminent extinctions. AB - Slowing rates of global biodiversity loss requires preventing species extinctions. Here we pinpoint centers of imminent extinction, where highly threatened species are confined to single sites. Within five globally assessed taxa (i.e., mammals, birds, selected reptiles, amphibians, and conifers), we find 794 such species, three times the number recorded as having gone extinct since 1500. These species occur in 595 sites, concentrated in tropical forests, on islands, and in mountainous areas. Their taxonomic and geographical distribution differs significantly from that of historical extinctions, indicating an expansion of the current extinction episode beyond sensitive species and places toward the planet's most biodiverse mainland regions. Only one-third of the sites are legally protected, and most are surrounded by intense human development. These sites represent clear opportunities for urgent conservation action to prevent species loss. PMID- 16344486 TI - Stereostructure of luminamicin, an anaerobic antibiotic, via molecular dynamics, NMR spectroscopy, and the modified Mosher method. AB - The absolute stereostructure of luminamicin, an anaerobic antibiotic, has been determined by using conformational analysis via high-temperature molecular dynamics, NMR spectroscopy, and the modified Mosher method. It was found that luminamicin has the S, S, R, R, R, R, S, S, S, R, and S configurations at C2, C4, C7, C9, C10, C11, C12, C13, C16, C28, and C29, respectively. This configuration is the same as that found in nodusmicin, which has a chemical structure quite similar to luminamicin. The structure of luminamicin consists of three different rings, i.e., a decalin ring, a 10-membered macrolactone ring, and a 14-membered macrolactone ring. The resulting three-dimensional structure of luminamicin shows an interesting feature in that the maleic anhydride functionality in conjugation with the enol ether group of the 14-membered macrolactone is nearly perpendicular to the plane of the other two rings. PMID- 16344487 TI - Discovery of an ornithurine bird and its implication for Early Cretaceous avian radiation. AB - An ornithurine bird, Hongshanornis longicresta gen. et sp. nov., represented by a nearly complete and articulated skeleton in full plumage, has been recovered from the lacustrine deposits of the Lower Cretaceous Jehol Group in Inner Mongolia, northeast China. The bird had completely reduced teeth and possessed a beak in both the upper and lower jaws, representing the earliest known beaked ornithurine. The preservation of a predentary bone confirms that this structure is not unique to ornithischian dinosaurs but was common in early ornithurine birds. This small bird had a strong flying capability with a low aspect ratio wing. It was probably a wader, feeding in shallow water or marshes. This find confirms that the aquatic environment had played a key role in the origin and early radiation of ornithurines, one branch of which eventually gave rise to extant birds near the Cretaceous/Tertiary boundary. This discovery provides important information not only for studying the origin and early evolution of ornithurines but also for understanding the differentiation in morphology, body size, and diet of the Early Cretaceous birds. PMID- 16344488 TI - Gap junctional hemichannel-mediated ATP release and hearing controls in the inner ear. AB - Connexin gap junctions play an important role in hearing function, but the mechanism by which this contribution occurs is unknown. Connexins in the cochlea are expressed only in supporting cells; no connexin expression occurs in auditory sensory hair cells. A gap junctional channel is formed by two hemichannels. Here, we show that connexin hemichannels in the cochlea can release ATP at levels that account for the submicromolar concentrations measured in the cochlear fluids in vivo. The release could be increased 3- to 5-fold by a reduction of extracellular Ca2+ or an increase in membrane stress, and blocked by gap junctional blockers. We also demonstrated that extracellular ATP at submicromolar levels apparently affected outer hair cell (OHC) electromotility, which is an active cochlear amplifier determining cochlear sensitivity to sound stimulation in mammals. ATP reduced OHC electromotility and the slope factor of the voltage dependence and shifted the operating point to reduce the active amplifier gain. ATP also reduced the generation of distortion products. Immunofluorescent staining showed that purinergic receptors P2x2 and P2x7 were distributed on the OHC surface. Blockage of P2 receptors eliminated the effect of ATP on the OHC electromotility. The data revealed that there is a hemichannel-mediated, purinergic intercellular signaling pathway between supporting cells and hair cells in the cochlea to control hearing sensitivity. The data also demonstrated a potential source of ATP in the cochlea. PMID- 16344489 TI - Sequence determinants of a conformational switch in a protein structure. AB - The Arc repressor of bacteriophage P22 is a dimeric member of the ribbon-helix helix family of transcription factors. Residues 9-14 of each wild-type Arc subunit pair to form two antiparallel beta-strands and have the alternating pattern of polar and nonpolar residues expected for a beta-ribbon with one solvent-exposed face and one face that forms part of the hydrophobic core. Simultaneously switching Asn-11 to Leu and Leu-12 to Asn changes the local binary sequence pattern to that of an amphipathic helix. Previous studies have shown that this double mutation results in replacement of the wild-type beta-ribbon by two right-handed 3(10)-helices. Moreover, an Arc variant bearing just the Asn-11 -> Leu mutation has an ambiguous binary pattern and can form either the ribbon or the helical structures, which interchange rapidly. Here, we study Arc mutants in which position 11 is occupied by Gly, Ala, Val, Ile, Leu, Met, Phe, or Tyr. These mutants adopt the wild-type beta-ribbon structure in a sequence context that stabilizes this fold, but they assume the alternative helical structure in a sequence background in which the wild-type fold is precluded by negative design. In an otherwise wild-type sequence background, the detailed chemical properties of the position 11 side chain dictate which of the two competing conformational folds is preferred. PMID- 16344490 TI - EULAR expects further development in 2006. PMID- 16344491 TI - Infectious origin of the antiphospholipid syndrome. PMID- 16344492 TI - Advanced glycation end product modification of bone proteins and bone remodelling: hypothesis and preliminary immunohistochemical findings. AB - BACKGROUND: The process of bone remodelling is disturbed in the development of osteoporosis. OBJECTIVE: To investigate if proteins in osteoporotic bone are modified by advanced glycation end products (AGEs), and whether these alterations are related to measures of bone remodelling based on histomorphometric findings. METHODS: Bone specimens taken from the iliac crest by bone biopsy of eight osteoporotic patients were investigated by histomorphometry and by immunohistochemical staining with the AGEs imidazolone and N(epsilon) carboxymethyllysine. RESULTS: Both AGEs were found in all bone specimens. The intensity of staining correlated with patient age. The percentage of bone surface covered with osteoblasts showed a significantly negative correlation with the staining intensity of both AGEs. CONCLUSIONS: It is known that AGEs can regulate proliferation and differentiation of osteoblastic cells and that AGE-specific binding sites are present in cultured osteoblast-like cells. Moreover, AGE induced biological effects in these cells might be mediated by RAGE (receptor of AGE) or by other AGE receptors in different stages of osteoblast development. The inverse relation between AGE staining intensity and the percentage of bone surface covered with osteoblasts in the trabecular bone may provide evidence that AGE modification of bone proteins disturbs bone remodelling. PMID- 16344493 TI - Expression of the matrix receptor CD44v5 on chondrocytes changes with osteoarthritis: an experimental investigation in the rabbit. AB - OBJECTIVE: To evaluate the expression of CD44v5 on chondrocytes of hyaline cartilage during the course of osteoarthritis (OA). METHODS: In 12 white New Zealand rabbits the anterior cruciate ligament (ACL) was resected to create an anterior instability of the knee. In 12 control rabbits only a sham operation without resection of the ACL was done. Four animals of each group were killed at 3, 6, and 12 weeks. The loadbearing area was evaluated histologically according to Mankin and by immunostaining for CD44v5. RESULTS: In the trial group, histological grades of OA showed a positive linear correlation with the time after surgery. Immunostaining showed an increased expression of CD44v5 in the control group after 3 and 6 weeks, which dropped to normal after 12 weeks. There was no difference between control and trial groups after 3 and 6 weeks, but a difference was seen after 12 weeks. A significant positive correlation between CD44v5 expression and the histological grade of OA was found (r = 0.314). CONCLUSIONS: An in vivo increase of expression of the hyaluronan receptor CD44v5 occurs during the course of OA. Further studies are needed to evaluate whether this pattern applies to man and whether new treatment approaches might evolve from this knowledge. PMID- 16344494 TI - Prevalence of the antiphospholipid syndrome in primary systemic vasculitis. AB - BACKGROUND: The antiphospholipid (APS or Hughes') syndrome, anticardiolipin antibodies (aCL), and the lupus anticoagulant (LA) are associated with systemic lupus erythematosus, malignancy, infection, and drugs. It has been described in patients with primary systemic vasculitis (PSV). OBJECTIVE: To determine the prevalence of APS in patients with PSV attending a vasculitis clinic and the prevalence of patients with positive aCL and/or the LA who do not fulfil the classification criteria for APS. METHODS: All case notes of patients attending the vasculitis clinic over a 12 month period were reviewed. Outpatients and inpatients were both included and were assessed for features of the APS and presence of aPL. Patients with positive aCL or LA tests were classified according to the significance of these results. RESULTS: Of 144 patients with PSV, 25 had positive aCL or LA on at least one occasion, representing a point prevalence of 17%. Of these, nine had definite APS (classified by the Sapporo criteria) and a further four patients had clinical and serological features of APS, although insufficient to satisfy the Sapporo criteria. Twelve had only positive aPL. CONCLUSION: The antiphospholipid syndrome, aCL, and the LA may occur in association with PSV. PMID- 16344495 TI - Immunological consequences of thalidomide treatment in Sjogren's syndrome. AB - OBJECTIVE: To study the immunological consequences of systemic thalidomide treatment in patients with Sjogren's syndrome. METHODS: Cytokine (tumour necrosis factor alpha (TNFalpha), interleukin (IL) 6) and soluble receptor (sIL2R) levels were measured in patient and control plasma (n = 7), before and after thalidomide treatment. Peripheral blood mononuclear cells were examined by FACS analysis for potential changes in specific cell populations (T cells, B cells, monocytes), and for the expression of activation markers (CD25, HLA-DR), costimulatory molecules (CD40, CD40L), TNF receptors, chemokine receptors, and adhesion molecules (L selectin (L-sel)). RESULTS: Owing to adverse effects of thalidomide, the treatment interval was limited. None the less, statistically significant changes in markers of cell activation were recorded in the four treated patients. Before treatment, HLA-DR, TNFRI, CXCRI, and CXCRII were raised in the patients compared with healthy controls (p<0.05) and their expression was down regulated after treatment. B cell numbers and expression of the adhesion molecule L-sel also declined with thalidomide. CONCLUSION: Significant changes in measures of cell activation were detected during thalidomide treatment within this limited study, which upon further investigation may offer insight into the underlying immunoregulatory pathways of thalidomide. PMID- 16344496 TI - Modification of hypertension and hypercholesterolaemia in patients with systemic lupus erythematosus: a quality improvement study. AB - BACKGROUND: Hypercholesterolaemia and hypertension are risk factors for coronary artery disease in patients with systemic lupus erythematosus (SLE). OBJECTIVE: To examine the recognition and management of hypercholesterolaemia and hypertension in patients with SLE before and after a quality improvement study. METHOD: Patients with SLE have been followed up prospectively at the University of Toronto Lupus Clinic since 1970. The charts of all patients who entered the clinic since 1990 were reviewed to ensure completeness of data on anti hypertensive treatment (AHT) and lipid lowering agents (LLA). Recognition and management of hypercholesterolaemia and hypertension were evaluated for the periods 1990-1995 and 1996-2001. RESULTS: Comparison of treatment between time periods showed that during 1990-1995 204/559 (36%) patients seen were hypertensive, of whom 180 (88%) were receiving AHT, and during 1996-2001 241/576 (42%) patients seen were hypertensive, of whom 232 (96%) were receiving AHT (p = 0.0013). A comparison of treatment for hypercholesterolaemia showed that 21/236 (9%) hypercholesterolaemic patients were being treated with LLA in the earlier period compared with 74/261 (28%) in the later period (p<0.0001). CONCLUSIONS: Treatment for hypertension and hyperlipidaemia has increased in the past 6 years compared with the previous 6 years, but a number of patients eligible for these treatments remain untreated. PMID- 16344497 TI - Rapid multifocal chondrolysis after liver transplantation in four patients. AB - BACKGROUND: Favourable clinical outcomes achieved with liver transplantation may be jeopardised by corticosteroid-induced osteoarticular complications, such as osteoporosis or aseptic osteonecrosis. CASE REPORT: A previously unreported joint complication in liver transplant recipients is described. METHODS: Retrospective study of liver transplant recipients in whom destructive joint disease developed, identified among 330 symptomatic patients out of a total of 1260 managed over a 15 year period (November 1984-January 2000) in a university based liver transplantation centre. RESULTS: In four patients (23-56 years), rapid chondrolysis developed 2-12 years after transplantation in more than three weightbearing or non-weightbearing joints free of radiological features of avascular osteonecrosis, evidence of infection, or defined inflammatory arthritis. Pathological examination of five hip joints and one knee joint found no evidence of infection or inflammation. Six joint replacement procedures were performed successfully in three patients. CONCLUSION: Clinicians managing liver transplant recipients, and perhaps recipients of other organs, should be aware that rapid chondrolysis may develop. The pathogenesis of this rare complication is unclear. PMID- 16344498 TI - Serum levels of monocyte chemotactic protein-3/CCL7 are raised in patients with systemic sclerosis: association with extent of skin sclerosis and severity of pulmonary fibrosis. AB - OBJECTIVE: To determine serum levels of monocyte chemotactic protein-3 (MCP-3) and its clinical associations in patients with systemic sclerosis (SSc). METHODS: Serum MCP-3 levels from 69 patients with SSc were examined by ELISA. RESULTS: Serum MCP-3 levels were raised in patients with SSc (n = 69) compared with healthy controls (n = 28). Patients with diffuse cutaneous SSc (n = 36) had higher levels of serum MCP-3 than those with limited cutaneous SSc (n = 33). Patients with raised MCP-3 levels had pulmonary fibrosis and decreased vital capacity (VC) more often than those with normal MCP-3 levels. MCP-3 levels correlated positively with the extent of skin fibrosis, and inversely with %VC and carbon monoxide transfer factor (Tlco) in patients with SSc. CONCLUSION: MCP 3 levels were increased in patients with SSc, and correlated with the extent of skin sclerosis and the severity of pulmonary fibrosis. These results suggest that MCP-3 may have a role in the development of fibrosis in SSc. PMID- 16344499 TI - Sexual dimorphism, but not testosterone itself, is responsible for ankylosing enthesitis of the ankle in B10.BR (H-2k) male mice. AB - BACKGROUND: Ankylosing enthesopathy (ANKENT) with progressive stiffening of ankle and tarsal joints of the hind limbs is a naturally occurring arthropathy in B10.BR mice. Some features are similar to those of the spondyloarthropathies in humans. OBJECTIVE: To study the role of sexual dimorphism and testosterone in the development of ANKENT. METHODS: The incidence of ANKENT was observed in non castrated, castrated, and testosterone substituted castrated male mice, and in control and testosterone treated female mice. RESULTS: ANKENT occurred only in males; it did not develop in males castrated at age 2-3 months but occurred in castrated males injected with testosterone. Females injected with testosterone did not develop ANKENT. CONCLUSION: Testosterone can replace what castration eliminates, at least in the postpubertally castrated males, but is itself not sufficient to induce joint disease. PMID- 16344500 TI - The presence of anti-cyclic citrullinated peptide antibody is associated with magnetic resonance imaging detection of bone marrow oedema in early stage rheumatoid arthritis. PMID- 16344501 TI - Early prediction of rheumatoid arthritis by serological variables and magnetic resonance imaging of the wrists and finger joints: results from prospective clinical examination. PMID- 16344502 TI - Brain natriuretic peptide is a potentially useful screening tool for the detection of cardiovascular disease in patients with rheumatoid arthritis. PMID- 16344503 TI - Severe relapse of Wegener's granulomatosis during the early postpartum period. PMID- 16344504 TI - HLA-DR11 and HLA-DR2 are negatively associated with autoantibody production in chronic hepatitis C. PMID- 16344505 TI - Gustatory sweating: Frey syndrome. PMID- 16344506 TI - Self-mutilation in the Lesch-Nyhan syndrome. PMID- 16344507 TI - Neurologic autoantibodies: cautious enthusiasm. PMID- 16344508 TI - Stroke thrombolysis in the elderly: risk or benefit? PMID- 16344509 TI - Has amobarbital expired? Considering the future of the Wada. PMID- 16344510 TI - Adult botulism type F in the United States, 1981-2002. AB - BACKGROUND: Clostridium botulinum neurotoxin types A, B, and E cause most cases of the paralytic disease botulism. Little is known about the epidemiology, clinical features, or microbiology of botulism type F. METHODS: Cases of adult type F botulism were identified by review of data collected by CDC's National Botulism Surveillance System between 1981 and 2002. A case was either an individual whose serum or stool demonstrated type F toxin or whose stool culture yielded an organism producing toxin type F. A detailed review of cases' medical charts and laboratory data from CDC and local health departments was performed. RESULTS: Between 1981 and 2002, 1,269 cases of botulism among adults and infants were reported to CDC; 13 (1%) were adult type F. The median age of type F cases was 54 years; 7 (54%) were female. None were part of outbreaks. A toxigenic Clostridium baratii was identified in 9 (69%) of 13 cases. Among 11 cases for which clinical data were available, all required mechanical ventilation for a median duration of 17 days (range, 10 to 84); 8 (73%) were intubated within 24 hours of symptom onset. All patients had nearly complete or complete quadriplegia at the nadir of neurologic dysfunction, which occurred on average on day 5. On average by day 8, improvement in neuromuscular function was noted. The median duration of acute hospitalization was 31 days (range, 20 to 60). No deaths were reported. In only one case was a possible foodborne etiology identified. CONCLUSIONS: Toxigenic C baratii are the sole documented causes of type F botulism in the United States since 1981. These cases are characterized by a fulminant course with rapid progression to respiratory failure and paralysis, making early recognition and intervention critical to appropriate management. PMID- 16344511 TI - Regional cerebral blood flow in Parkinson disease with nonpsychotic visual hallucinations. AB - BACKGROUND: Patients with Parkinson disease (PD) often experience visual hallucinations (VH) with retained insight (nonpsychotic) but the precise mechanism remains unclear. OBJECTIVE: To clarify which neural substrates participate in nonpsychotic VH in PD, the authors evaluated regional cerebral blood flow (rCBF) changes in patients with PD and VH. METHODS: The authors compared 24 patients with PD who had nonpsychotic VH (hallucinators) and 41 patients with PD who had never experienced VH (non-hallucinators) using SPECT images with N-isopropyl-p-[(123)I]iodoamphetamine. There were no significant differences in age, sex, duration of disease, doses of PD medications, Hoehn and Yahr scale, or Mini-Mental State Examination (MMSE) scores between the two groups. The rCBF data were analyzed using statistical parametric mapping (SPM). RESULTS: The rCBF in the right fusiform gyrus was lower in the hallucinators than in the non-hallucinators (corrected p < 0.05 at cluster levels). The hallucinators revealed higher rCBF in the right superior and middle temporal gyri than the non-hallucinators (uncorrected p < 0.001). These significant differences were demonstrated after MMSE scores and duration of disease, which are the relevant factors associated with VH, were covariated out. CONCLUSIONS: Nonpsychotic visual hallucinations in Parkinson disease (PD) may be associated with hypoperfusion in the right fusiform gyrus and hyperperfusion in the right superior and middle temporal gyri. These temporal regions are important for visual object recognition and these regional cerebral blood flow changes are associated with inappropriate visual processing and are responsible for nonpsychotic visual hallucinations in PD. PMID- 16344512 TI - Dementia in Parkinson disease: functional imaging of cholinergic and dopaminergic pathways. AB - OBJECTIVE: To assess neurochemical deficits in patients with Parkinson disease (PD) associated dementia (PDD) in vivo. METHODS: The authors performed combined PET with N-[11C]-methyl-4-piperidyl acetate (MP4A) and 18F-fluorodopa (FDOPA) for evaluation of cholinergic and dopaminergic transmitter changes in 17 non-demented patients with PD and 10 patients with PDD. Data were compared to 31 age-matched controls by a combined region-of-interest and voxel-based Statistical Parametric Mapping analysis. RESULTS: The striatal FDOPA uptake was significantly decreased in PD and PDD without differences between the groups. The global cortical MP4A binding was severely reduced in PDD (29.7%, p < 0.001 vs controls) and moderately decreased in PD (10.7%, p < 0.01 vs controls). The PDD group had lower parietal MP4A uptake rates than did patients with PD. Frontal and temporo-parietal cortices showed a significant covariance of striatal FDOPA reduction and decreased MP4A binding in patients with PDD. CONCLUSIONS: While non-demented patients with Parkinson disease had a moderate cholinergic dysfunction, subjects with Parkinson disease associated dementia (PDD) presented with a severe cholinergic deficit in various cortical regions. The finding of a closely associated striatal FDOPA and cortical MP4A binding reduction suggests a common disease process leading to a complex transmitter deficiency syndrome in PDD. PMID- 16344513 TI - Etomidate speech and memory test (eSAM): a new drug and improved intracarotid procedure. AB - BACKGROUND: The intracarotid amobarbital procedure (IAP) is an important part of comprehensive investigation of patients who are candidates for surgical treatment of epilepsy. Owing to repeated and lengthy shortages of amobarbital, causing delays in elective surgery, attempts have been made to find a suitable alternative anesthetic. The authors report their experience using etomidate, a widely used agent for the induction of anesthesia. METHODS: Sixteen consecutive patients requiring IAP to evaluate memory or to lateralize speech underwent the procedure using etomidate. Prior to the procedure a catheter was placed in the internal carotid artery and an angiogram was performed. EEG was recorded and read online by an electroencephalographer. An anesthetist injected the drug, administered by bolus followed by an infusion, which was maintained until each speech measure had been sampled and new memory items had been introduced. The infusion was then stopped and testing continued as in a standard IAP. RESULTS: In all cases (30 hemispheres) contralateral hemiplegia followed injection. EEG slow waves were observed in every injected hemisphere, with some contralateral slowing anteriorly in 18. Global aphasia with preserved attention and cooperation followed dominant-hemisphere injections. These phenomena remained during infusion, and upon its termination returned gradually to baseline over a period of about 4 minutes. CONCLUSIONS: Etomidate is a viable alternative to amobarbital, and its administration by bolus followed by infusion offers an improvement over the traditional intracarotid amobarbital procedure. Cognitive tests can be performed during an assured hemianesthesia of the injected hemisphere. PMID- 16344514 TI - Serum antibodies in epilepsy and seizure-associated disorders. AB - OBJECTIVE: To investigate whether autoantibodies to ion channels and other neural antigens are present in the sera of patients with epilepsy and seizure-related diseases. METHODS: Sera were obtained from 139 patients, including 26 with preexisting autoimmune disease, 46 in whom an autoimmune basis was suspected, and 67 with drug-resistant epilepsy. The sera were assayed for antibodies to voltage gated potassium (VGKC) and calcium (VGCC) channels, glutamic acid decarboxylase (GAD), gangliosides, glutamate receptor type 3, cardiolipins, DNA, and nuclear antigens; the results were compared with results from a large cohort of healthy and disease controls. RESULTS: Increased titers of VGKC antibodies (>100 pM) were detected in 16 of 139 (11%) patients with seizures but only 1 control (0.5%). Eight VGKC-positive patients presented with an acute/subacute illness, and 5 of these had the highest VGKC antibodies; 3 patients improved spontaneously, another 5 patients responded well to immunomodulatory therapy. The other VGKC-positive patients had longer disease duration (>6 years) and intermediate levels of antibodies; immunotherapies have not been tested in this group. Very high levels of GAD antibodies (>1,000 U) were found in an additional 3 patients (2.1%) with long-standing drug-resistant epilepsy. CONCLUSIONS: The presence of autoantibodies to voltage-gated potassium channels and glutamic acid decarboxylase suggests that the immune system may contribute to certain forms of epilepsy or seizure-associated disorders. Further studies are needed to determine whether the antibodies are pathogenic. PMID- 16344515 TI - Double-blind, placebo-controlled study of lamotrigine in primary generalized tonic-clonic seizures. AB - OBJECTIVE: To evaluate the efficacy and tolerability of adjunctive lamotrigine in primary generalized tonic-clonic (PGTC) seizures in a randomized, double-blind, placebo-controlled trial. METHODS: Patients with a diagnosis of epilepsy with PGTC seizures who were receiving one or two antiepileptic drugs at study entry were eligible. Patients with partial seizures were excluded on the basis of seizure history and screening EEGs. The study comprised a baseline phase, an escalation phase during which study medication was titrated to a target dose, and a 12-week maintenance phase during which doses of lamotrigine/placebo and concomitant antiepileptic drugs were maintained. RESULTS: Of the 121 randomized patients ages 2 to 55 years, 117 (58 lamotrigine, 59 placebo) entered the escalation phase and received study medication. During the escalation and maintenance phases combined, median percent reduction in PGTC seizure frequency was 66.5% with lamotrigine compared with 34.2% with placebo (p = 0.006). The corresponding numbers for lamotrigine and placebo were 60.6% and 32.8% (p = 0.038) during the escalation phase and 81.9% and 43.0% (p = 0.006) during the maintenance phase. During the maintenance phase, 72% of lamotrigine-treated patients compared with 49% of placebo-treated patients experienced a > or = 50% reduction in frequency of PGTC seizures (p = 0.014). A similar pattern of results was observed for all generalized seizures. The most common drug-related adverse events were dizziness (5% lamotrigine, 2% placebo), somnolence (5% lamotrigine, 2% placebo), and nausea (5% lamotrigine, 3% placebo). CONCLUSIONS: Adjunctive lamotrigine is effective in the treatment of primary generalized tonic-clonic seizures and has a favorable tolerability profile. PMID- 16344516 TI - Changes in depression and anxiety after resective surgery for epilepsy. AB - OBJECTIVE: To determine changes in depression and anxiety after resective surgery. METHODS: Data from subjects enrolled in a prospective multicenter study of resective epilepsy surgery were reviewed with the Beck Psychiatric Symptoms Scales (Beck Depression Inventory [BDI] and Beck Anxiety Inventory [BAI]) and Composite International Diagnostic Interview (CIDI) up to a 24-month period. chi2 analyses were used to correlate proportions. RESULTS: A total of 358 presurgical BDI and 360 BAI results were reviewed. Moderate and severe levels of depression were reported in 22.1% of patients, and similar levels of anxiety were reported by 24.7%. Postoperative rates of depression and anxiety declined at the 3-, 12-, and 24-month follow-up periods. At the 24-month follow-up, moderate to severe levels of depression symptoms were reported in 17.6 and 14.7% of the patients who continued to have postoperative seizures. Moderate to severe depression and anxiety were found in 8.2% of those who were seizure-free. There was no relationship, prior to surgery, between the presence or absence of depression and anxiety and the laterality or location of the seizure onset. There were no significant relationships between depression or anxiety at 24-month follow-up and the laterality or location of the surgery. CONCLUSIONS: Depression and anxiety in patients with refractory epilepsy significantly improve after epilepsy surgery, especially in those who are seizure-free. Neither the lateralization nor the localization of the seizure focus or surgery was associated with the risk of affective symptoms at baseline or after surgery. PMID- 16344517 TI - Do generalized tonic-clonic seizures in infancy exist? AB - OBJECTIVE: To determine the frequency of generalized tonic-clonic seizures (GTCS) in infants (1 month to 2 years). METHODS: From a total of 2,112 patients monitored in our video-EEG lab from May 2000 through January 2005, 109 distinct seizures in 77 infants were reviewed. Eight events in eight patients were excluded because of video files insufficiently reliable to determine the clinical characteristics with precision. The clinical manifestations and electrographic features of the remaining 101 seizures in 69 infants were retrospectively analyzed. RESULTS: The authors did not observe a single GTCS. Four patients had icti that resembled GTCS, but careful analysis of these episodes revealed that three of them had a focal onset and that the fourth had a slightly different sequence of events. CONCLUSIONS: Generalized tonic-clonic seizures are rarely, if ever, seen in infants younger than age 2 in a tertiary-care pediatric epilepsy unit. Instead, they more commonly occur in older children, particularly in the well-characterized epilepsy syndromes of childhood and adolescence. PMID- 16344518 TI - Peroneal intraneural ganglion. PMID- 16344519 TI - Neuropsychological evidence of modular organization of the near peripersonal space. AB - BACKGROUND: The space closely surrounding the body, near peripersonal space, is represented through multisensory coding, as witnessed by the spatial distribution of visual-tactile extinction in brain-damaged patients. Typically, tactile stimuli delivered on the contralesional hand are more severely extinguished by visual stimuli presented near to than far from the ipsilesional hand. OBJECTIVE: To investigate whether the near peripersonal space is a unitary multisensory representation of the peribody space or is organized in a modular fashion, separately representing different body parts. METHODS: Cross-modal extinction was used to assess right brain-damaged patients with left tactile extinction by presenting a combination of ipsilesional visual and contralesional tactile stimuli, both between homologous body parts (i.e., the two hands and sides of the face) and between nonhomologous body parts (i.e., right hand + left face and right face + left hand). RESULTS: Visual-tactile extinction observed in the near peripersonal space of homologous body parts was more severe than that obtained between nonhomologous body parts. In contrast, cross-modal extinction observed in the far peripersonal space was overall weak and comparable when stimulating homologous or nonhomologous body sectors. In addition, a clear near-far modulation of visual-tactile extinction was obtained only when stimulating homologous, but not nonhomologous, body parts. These findings were observed irrespective of the severity of unisensory tactile extinction. CONCLUSIONS: Near peripersonal space is not unitary but is functionally organized in modules and consists of at least two different spatial representations, one around the hand and another around the face, which might be selectively affected in patients with extinction. PMID- 16344520 TI - Predictors of relapse rate in MS clinical trials. AB - BACKGROUND: The annual relapse rate has been commonly used as a primary efficacy endpoint in phase III multiple sclerosis (MS) clinical trials. The aim of this study was to determine the relative contribution of different possible prognostic factors available at baseline to the on-study relapse rate in MS. METHODS: A total of 821 patients from the placebo arms of the Sylvia Lawry Centre for Multiple Sclerosis Research (SLCMSR) database were available for this analysis. The univariate relationships between on-study relapse rate and the baseline demographic, clinical, and MRI-based predictors were assessed. The multiple relationships were then examined using a Poisson regression model. Two predictor subsets were selected. Subset 1 included age at disease onset, disease duration, sex, Expanded Disability Status Scale (EDSS) at baseline, number of relapses in the last 24 months prior to baseline, and the disease course (relapsing remitting [RR] and secondary progressive [SP]). Subset 2 consisted of Subset 1 plus gadolinium enhancement status in MRI. The number of patients for developing the models with no missing values was 727 for Subset 1 and 306 for Subset 2. RESULTS: The univariate relationships show that the on-study relapse rate was higher for younger and for female patients, for RR patients than for SP patients, and for patients with positive enhancement status at entry (Wilcoxon test, p < 0.05). A higher on-study relapse rate was associated with a shorter disease duration, lower entry EDSS, more pre-study relapses, and more enhancing lesions in T1 at entry. The fitted Poisson model shows that disease duration (estimate = -0.02) and previous relapse number (estimate = 0.59 for one, 0.91 for two, and 1.45 for three or more relapses vs no relapses) remain. The authors were able to confirm these findings in a second, independent dataset. CONCLUSIONS: The relapse number prior to entry into clinical trials together with disease duration are the best predictors for the on-study relapse rate. Disease course did not contribute independently because its effect is covered by the pre-study relapse rate. Gadolinium enhancement status, given the other covariates, has no significant influence on the on-study relapse rate. PMID- 16344521 TI - The effects of executive and behavioral dysfunction on the course of ALS. AB - OBJECTIVE: To determine whether patients with ALS-frontotemporal lobar dementia (FTLD) have a shorter survival and are less compliant with recommended treatments than those with ALS who have normal executive and behavioral function (classic ALS). METHODS: Survival analysis from ALS symptom onset to death included 81 of 100 consecutive patients who could be classified definitely as ALS with abnormal executive or behavioral function or as classic ALS. Criteria were defined for compliance with noninvasive positive-pressure ventilation (NPPV) and percutaneous endoscopic gastrostomy (PEG). RESULTS: Median survival was 2 years 4 months for the 28 patients with FTLD and 3 years 3 months for the 53 patients with classic ALS (relative hazard for death 1.93, CI 1.09 to 3.43; p = 0.024). However, the relative hazard associated with FTLD (1.49) in the multivariate model was diminished by the association of FTLD with bulbar onset and older age and was not significant in this sample size. With bulbar onset, median survival was 2 years 0 months for the 14 with ALS-FTLD and 2 years 10 months for the 10 with classic ALS (relative hazard for death 2.78, CI 1.02 to 7.55; p = 0.045), and older age was not a significant risk. Noncompliance with NPPV and PEG were 75% and 72% in ALS FTLD, respectively, vs 38% and 31% in classic ALS (relative risks 2.00 and 2.34; p = 0.013 and 0.022). CONCLUSIONS: Survival is significantly shorter among patients with ALS-FTLD than with classic ALS. Furthermore, patients with ALS-FTLD are twice as likely to be noncompliant. PMID- 16344522 TI - Evaluation of a clinical screening tool for HIV-associated sensory neuropathies. AB - OBJECTIVE: To evaluate the performance characteristics of a brief clinical neuropathy screening tool for use in sensory neuropathies complicating HIV infection. METHODS: The authors assessed 80 patients using the Brief Peripheral Neuropathy Screen (BPNS). Patients were defined as having neuropathy if they had both symptoms and signs consistent with this diagnosis. All subjects underwent sensory threshold testing and lower limb epidermal nerve fiber quantification using punch skin biopsy as objective measures. RESULTS: Individuals defined as having neuropathy using the BPNS (n = 37) performed less well on sensory threshold testing than other HIV-infected individuals (p < 0.0001 for warming, cooling, and vibration) and also had lower distal calf epidermal nerve fiber densities (p < 0.0001). Individuals who had symptoms but no neuropathic signs (n = 13) did not perform differently on any objective testing compared with neuropathy-free individuals, supporting the decision to require signs as well as symptoms as an operational criterion for the diagnosis of neuropathy. Of the symptoms listed in the screening tool, the presence of numbness had the greatest diagnostic efficiency for identifying those with neuropathy. CONCLUSION: The Brief Neuropathy Screening Tool (and the chosen definition of neuropathy) accurately detects those HIV-infected individuals with the greatest degree of peripheral nerve dysfunction and pathology. This is a valid neuropathy screening tool for use in the context of HIV infection, and is simple enough to be applicable in resource-limited settings. PMID- 16344524 TI - Captain Cook on poison fish. AB - On his second voyage of discovery, Captain James Cook charted much of the South Pacific. The journey was long, from 1772 to 1775. During the exploration, the geographic, ethnographic, and scientific variety provided no shortage of work for the accompanying naturalists, astronomers, navigators, and painters. Culinary discoveries included new species of fish, many of which were sketched, dressed, and ultimately eaten. The examined journals and correspondence document clinical poisonings after ingestion of two different species of fish. The clinical findings are described and likely represent ciguatera and tetrodotoxin poisonings. Mechanisms of these toxin's actions are discussed in light of more recent studies. PMID- 16344523 TI - Plasma cells in muscle in inclusion body myositis and polymyositis. AB - BACKGROUND: Previous immunohistochemical studies of muscle from patients with inclusion body myositis and polymyositis found many more T cells than B cells, suggesting a role for intramuscular cell-mediated immune mechanisms rather than humoral mechanisms. METHODS: Microarray studies were performed on muscle biopsy specimens from 40 patients with inclusion body myositis (IBM; n = 23), polymyositis (PM; n = 6), and without neuromuscular disease (n = 11). Reverse transcription PCR of selected immunoglobulin gene transcripts was performed on two patient samples. Qualitative immunohistochemical studies for B-cell lineage cell surface markers were performed on 28 muscle specimens and quantitative studies performed on a subset of 19 untreated patients with IBM or PM. CD138+ cells were isolated from muscle using laser capture microdissection, and immunoglobulin transcripts were PCR amplified to determine the presence or absence of immunoglobulin gene rearrangements unique to the B-cell lineage. RESULTS: Immunoglobulin gene transcripts accounted for 59% in IBM and 33% in PM of the most stringently defined highest differentially expressed muscle transcripts compared with normal. Plasma cells, terminally differentiated B cells expressing CD138 but not CD19 or CD20, are present in IBM and PM muscle in numbers several times higher than B cells. CONCLUSIONS: There are differentiated B cells in the form of CD138+ plasma cells within the muscle of patients with inclusion body myositis and polymyositis. The principle of linked recognition of B-cell activation predicts several strategies for autoantigen discovery that could not otherwise be pursued through the study of the infiltrating T-cell population alone. PMID- 16344525 TI - Is hospitalization after TIA cost-effective on the basis of treatment with tPA? AB - A 24-hour hospitalization for TIA could be cost-effective simply by increasing the likelihood that patients will receive tissue plasminogen activator if a stroke occurs. The authors performed a cost-utility analysis of 24-hour hospitalization for patients diagnosed with recent TIA. The overall cost effectiveness ratio was 55,044 dollars per quality-adjusted life-year, a value considered borderline cost-effective. For patients with higher risk of stroke, admission was cost-effective. PMID- 16344526 TI - Alpha7-acetylcholine receptor antibodies in two patients with Rasmussen encephalitis. AB - Rasmussen encephalitis (RE) sera were screened for antibodies to human alpha7 nicotinic acetylcholine receptors (nAChRs) using electrophysiology, calcium imaging, and ligand binding assays. Sera from two of nine patients with RE blocked ACh-induced currents through alpha7 nAChRs and the ACh-induced rise in intracellular free calcium ([Ca2+]i) and inhibited (125)I-alpha-bungarotoxin binding in cells expressing alpha7 nAChRs. Thus, the alpha7 nAChR is a potential target for pathogenic antibodies in patients with RE. PMID- 16344527 TI - Complex motor behaviors in temporal lobe epilepsy. AB - Complex motor behaviors differing from typical automatisms were found in 12 of 502 patients with temporal lobe epilepsy. Movements involved proximal limb segments (6) or body axis (6) and were often preceded by auras and followed by automatisms. Seven of 12 patients are seizure free after surgery. The other 5 patients declined surgery. PMID- 16344528 TI - Leptin, ghrelin, and adiponectin in epileptic patients treated with valproic acid. AB - The authors studied 40 epileptic patients treated with valproate and 40 healthy controls for at least 2 years. At the end of follow-up, 15 epileptic patients (37.5%) had development of obesity. They showed circulating leptin and insulin levels significantly higher and ghrelin and adiponectin levels significantly lower than those of patients who did not gain weight. PMID- 16344529 TI - Low-glycemic-index treatment: a liberalized ketogenic diet for treatment of intractable epilepsy. AB - The ketogenic diet is often effective for intractable epilepsy, but many patients have trouble complying with the strict regimen. The authors tested an alternative diet regimen, a low-glycemic-index treatment, with more liberal total carbohydrate intake but restricted to foods that produce relatively little increase in blood glucose (glycemic index < 50). Ten of 20 patients treated with this regimen experienced a greater than 90% reduction in seizure frequency. PMID- 16344530 TI - MRI of trigeminal zoster. PMID- 16344531 TI - Comparison of family histories in FTLD subtypes and related tauopathies. AB - Pedigrees from 269 patients with frontotemporal lobar degeneration (FTLD), including frontotemporal dementia (FTD), FTD with ALS (FTD/ALS), progressive nonfluent aphasia, semantic dementia (SD), corticobasal degeneration, and progressive supranuclear palsy were analyzed to determine the degree of heritability of these disorders. FTD/ALS was the most and SD the least heritable subtype. FTLD syndromes appear to have different etiologies and recurrence risks. PMID- 16344532 TI - Two patients with COMT inhibitor-induced hepatic dysfunction and UGT1A9 genetic polymorphism. AB - The authors report two cases of catechol-O-methyltransferase (COMT) inhibitor induced asymptomatic hepatic dysfunction in women with Parkinson disease. The patients were genotyped for the UDP-glucuronosyltransferase (UGT) 1A9 gene (which encodes the main COMT inhibitor-metabolizing enzyme), and found to carry mutations leading to defective glucuronidation activity. This suggests that UGT1A9 poor metabolizer genotype(s) may be a predisposing factor for COMT inhibitor-induced hepatotoxicity. PMID- 16344533 TI - BDNF genetic variants are associated with onset age of familial Parkinson disease: GenePD Study. AB - Brain-derived neurotrophic factor (BDNF) stimulates neuronal growth and protects nigral dopamine neurons in animal models of Parkinson disease (PD). Therefore, BDNF is a candidate gene for PD. The authors investigated five single-nucleotide polymorphisms in 597 cases of familial PD. Homozygosity for the rare allele of the functional BDNF G196A (Val66Met) variant was associated with a 5.3-year older onset age (p = 0.0001). These findings suggest that BDNF may influence PD onset age. PMID- 16344534 TI - Familial basilar migraine associated with a new mutation in the ATP1A2 gene. AB - Basilar migraine (BM), familial hemiplegic migraine (FHM), and sporadic hemiplegic migraine (SHM) are phenotypically similar subtypes of migraine with aura, differentiated only by motor symptoms, which are absent in BM. Mutations in CACNA1A and ATP1A2 have been found in FHM. The authors detected a novel mutation in the ATP1A2 gene (R548H) in members of a family with BM, suggesting that BM and FHM may be allelic disorders. PMID- 16344535 TI - MRI detection of cysts of the knee causing common peroneal neuropathy. AB - In 10 consecutive patients with footdrop due to common peroneal neuropathy without an obvious cause, MRI of the knee showed pathology at the fibular head in 6, including 5 patients with clinically unsuspected cysts of the tibiofibular joint. All 6 of the patients improved with surgery. PMID- 16344536 TI - A family with McLeod syndrome and calpainopathy with clinically overlapping diseases. AB - The authors describe a family with six patients with muscular dystrophy with a variable course. One is a compound heterozygote for CAPN3 mutations (calpainopathy) and the others have a single CAPN3 mutation. Linkage analysis and sequencing revealed a XK gene mutation (McLeod syndrome). This illustrates the variable phenotype of XK mutations and suggests the possibility that CAPN3 heterozygotes may have their condition caused by nonallelic mutations in other unrelated genes. PMID- 16344537 TI - Tolerance of high-dose (3,000 mg/day) coenzyme Q10 in ALS. AB - An open-label dose-escalation trial was performed to assess the safety and tolerability of high doses of coenzyme Q10 (CoQ10) in ALS. CoQ10, a cofactor in mitochondrial electron transfer, may improve the mitochondrial dysfunction in ALS. In this study, CoQ10 was safe and well tolerated in 31 subjects treated with doses as high as 3,000 mg/day for 8 months. PMID- 16344538 TI - Saccade testing in the diagnosis and treatment of type 3 Gaucher disease. PMID- 16344539 TI - Dementia due to neurosyphilis with persistently negative CSF VDRL. PMID- 16344540 TI - Diaphragmatic flutter after spinal cord trauma in a child. PMID- 16344541 TI - Hypogeusia and hearing loss in a patient with an inferior collicular infarction. PMID- 16344542 TI - Cefepime- and cefixime-induced encephalopathy in a patient with normal renal function. PMID- 16344544 TI - fMRI reveals large-scale network activation in minimally conscious patients. PMID- 16344545 TI - Dopaminergic dysfunction in unrelated, asymptomatic carriers of a single parkin mutation. PMID- 16344546 TI - Reasons for exclusion from thrombolytic therapy following acute ischemic stroke. PMID- 16344547 TI - Episodic third nerve palsy with cryptococcal meningitis. PMID- 16344548 TI - The RASSF1A tumor suppressor activates Bax via MOAP-1. AB - The novel tumor suppressor RASSF1A is frequently inactivated during human tumorigenesis by promoter methylation. RASSF1A may serve as a node in the integration of signaling pathways controlling a range of critical cellular functions including cell cycle, genomic instability, and apoptosis. The mechanism of action of RASSF1A remains under investigation. We now identify a novel pathway connecting RASSF1A to Bax via the Bax binding protein MOAP-1. RASSF1A and MOAP-1 interact directly, and this interaction is enhanced by the presence of activated K-Ras. RASSF1A can activate Bax via MOAP-1. Moreover, activated K-Ras, RASSF1A, and MOAP-1 synergize to induce Bax activation and cell death. Analysis of a tumor derived point mutant of RASSF1A showed that the mutant was defective for the MOAP 1 interaction and for Bax activation. Moreover, inhibition of RASSF1A by shRNA impaired the ability of K-Ras to activate Bax. Thus, we identify a novel pro apoptotic pathway linking K-Ras, RASSF1A and Bax that is specifically impaired in some human tumors. PMID- 16344549 TI - Mechanisms for picrotoxin block of alpha2 homomeric glycine receptors. AB - It is well known that the convulsant alkaloid picrotoxin (PTX) can inhibit neuronal gamma-aminobutyric acid (GABA) and homomeric glycine receptors (GlyR). However, the mechanism for PTX block of alpha(2) homomeric GlyR is still unclear compared with that of alpha(1) homomeric GlyR, GABA(A), and GABA(C) receptors. Furthermore, PTX effects on GlyR kinetics have been poorly explored at the single channel level. Hence, we used the patch-clamp technique in the outside-out configuration to investigate the mechanism of PTX suppression of currents carried by alpha(2) homomeric GlyRs stably transfected into Chinese hamster ovary cells. PTX inhibited the alpha(2) homomeric GlyR current elicited by glycine in a concentration-dependent and voltage-independent manner. Both competitive and noncompetitive mechanisms were observed. PTX decreased the mean open time of the GlyR channel in a concentration-dependent manner, suggesting that PTX can block channel openings and bind to the receptor in the open channel conformation. When PTX and glycine were co-applied, a small rebound current was observed during drug washout. Application of PTX during the deactivation phase of glycine-induced currents eliminated the rebound current and accelerated the deactivation time course in a concentration-dependent manner. PTX could not bind to the unbound conformation of GlyR, but could be trapped at its binding site when the channel closed during glycine dissociation. Based on these observations, we propose a kinetic Markov model in which PTX binds to the alpha(2) homomeric GlyR in both the open channel state and the fully liganded closed state. Our data suggest a new allosteric mechanism for PTX inhibition of wild-type homomeric alpha(2) GlyR. PMID- 16344550 TI - Differential use of functional domains by coiled-coil coactivator in its synergistic coactivator function with beta-catenin or GRIP1. AB - beta-Catenin, a pivotal component of the Wnt-signaling pathway, binds to and serves as a transcriptional coactivator for the T-cell factor/lymphoid enhancer factor (TCF/LEF) family of transcriptional activator proteins and for the androgen receptor (AR), a nuclear receptor. Three components of the p160 nuclear receptor coactivator complex, including CARM1, p300/CBP, and GRIP1 (one of the p160 coactivators), bind to and cooperate with beta-catenin to enhance transcriptional activation by TCF/LEF and AR. Here we report that another component of the p160 nuclear receptor coactivator complex, the coiled-coil coactivator (CoCoA), directly binds to and cooperates synergistically with beta catenin as a coactivator for AR and TCF/LEF. CoCoA uses different domains to bind GRIP1 and beta-catenin, and it uses different domains to transmit the activating signal to the transcription machinery, depending on whether it is bound to GRIP1 or beta-catenin. CoCoA associated specifically with the promoters of transiently transfected and endogenous target genes of TCF/LEF, and reduction of the endogenous CoCoA level decreased the ability of TCF/LEF and beta-catenin to activate transcription of transient and endogenous target genes. Thus, CoCoA uses different combinations of functional domains to serve as a physiologically relevant component of the Wnt/beta-catenin signaling pathway and the androgen signaling pathway. PMID- 16344551 TI - Mutator phenotypes caused by substitution at a conserved motif A residue in eukaryotic DNA polymerase delta. AB - Eukaryotic DNA polymerase (Pol) delta replicates chromosomal DNA and is also involved in DNA repair and genetic recombination. Motif A in Pol delta, containing the sequence DXXXLYPSI, includes a catalytically essential aspartic acid as well as other conserved residues of unknown function. Here, we used site directed mutagenesis to create all 19 amino acid substitutions for the conserved Leu(612) in Motif A of Saccharomyces cerevisiae Pol delta. We show that substitutions at Leu(612) differentially affect viability, sensitivity to genotoxic agents, cell cycle progression, and replication fidelity. The eight viable mutants contained Ile, Val, Thr, Met, Phe, Lys, Asn, or Gly substitutions. Individual substitutions varied greatly in the nature and extent of attendant phenotypic deficiencies, exhibiting mutation rates that ranged from near wild type to a 37-fold increase. The L612M mutant exhibited a 7-fold elevation of mutation rate but essentially no detectable effects on other phenotypes monitored; the L612T mutant showed a nearly wild type mutation rate together with marked hypersensitivity to genotoxic agents; and the L612G and L612N strains exhibited relatively high mutation rates and severe deficits overall. We compare our results with those for homologous substitutions in prokaryotic and eukaryotic DNA polymerases and discuss the implications of our findings for the role of Leu(612) in replication fidelity. PMID- 16344552 TI - Glucose-stimulated DNA synthesis through mammalian target of rapamycin (mTOR) is regulated by KATP channels: effects on cell cycle progression in rodent islets. AB - The aim of this study was to define metabolic signaling pathways that mediate DNA synthesis and cell cycle progression in adult rodent islets to devise strategies to enhance survival, growth, and proliferation. Since previous studies indicated that glucose-stimulated activation of mammalian target of rapamycin (mTOR) leads to [3H]thymidine incorporation and that mTOR activation is mediated, in part, through the K(ATP) channel and changes in cytosolic Ca2+, we determined whether glyburide, an inhibitor of K(ATP) channels that stimulates Ca2+ influx, modulates [3H]thymidine incorporation. Glyburide (10-100 nm) at basal glucose stimulated [3H]thymidine incorporation to the same magnitude as elevated glucose and further enhanced the ability of elevated glucose to increase [3H]thymidine incorporation. Diazoxide (250 microm), an activator of KATP channels, paradoxically potentiated glucose-stimulated [3H]thymidine incorporation 2-4-fold above elevated glucose alone. Cell cycle analysis demonstrated that chronic exposure of islets to basal glucose resulted in a typical cell cycle progression pattern that is consistent with a low level of proliferation. In contrast, chronic exposure to elevated glucose or glyburide resulted in progression from G0/G1 to an accumulation in S phase and a reduction in G2/M phase. Rapamycin (100 nm) resulted in an approximately 62% reduction of S phase accumulation. The enhanced [3H]thymidine incorporation with chronic elevated glucose or glyburide therefore appears to be associated with S phase accumulation. Since diazoxide significantly enhanced [3H]thymidine incorporation without altering S phase accumulation under chronic elevated glucose, this increase in DNA synthesis also appears to be primarily related to an arrest in S phase and not cell proliferation. PMID- 16344553 TI - Amyloid precursor-like protein 1 influences endocytosis and proteolytic processing of the amyloid precursor protein. AB - Ectodomain shedding of the amyloid precursor protein (APP) is a key regulatory step in the generation of the Alzheimer disease amyloid beta peptide (Abeta). The molecular mechanisms underlying the control of APP shedding remain little understood but are in part dependent on the low density lipoprotein receptor related protein (LRP), which is involved in APP endocytosis. Here, we show that the APP homolog APLP1 (amyloid precursor-like protein 1) influences APP shedding. In human embryonic kidney 293 cells expression of APLP1 strongly activated APP shedding by alpha-secretase and slightly reduced beta-secretase cleavage. As revealed by domain deletion analysis, the increase in APP shedding required the NPTY amino acid motif within the cytoplasmic domain of APLP1. This motif is conserved in APP and is essential for the endocytosis of APP and APLP1. Unrelated membrane proteins containing similar endocytic motifs did not affect APP shedding, showing that the increase in APP shedding was specific to APLP1. In LRP deficient cells APLP1 no longer induced APP shedding, suggesting that in wild type cells APLP1 interferes with the LRP-dependent endocytosis of APP and there by increases APP alpha-cleavage. In fact, an antibody uptake assay revealed that expression of APLP1 reduced the rate of APP endocytosis. In summary, our study provides a novel mechanism for APP shedding, in which APLP1 affects the endocytosis of APP and makes more APP available for alpha-secretase cleavage. PMID- 16344554 TI - A missense mutation in the bovine SLC35A3 gene, encoding a UDP-N acetylglucosamine transporter, causes complex vertebral malformation. AB - The extensive use of a limited number of elite bulls in cattle breeding can lead to rapid spread of recessively inherited disorders. A recent example is the globally distributed syndrome Complex Vertebral Malformation (CVM), which is characterized by misshapen and fused vertebrae around the cervico-thoracic junction. Here, we show that CVM is caused by a mutation in the Golgi-resident nucleotide-sugar transporter encoded by SLC35A3. Thus, the disease showed complete cosegregation with the mutation in a homozygous state, and proteome patterns indicated abnormal protein glycosylation in tissues of affected animals. In addition, a yeast mutant that is deficient in the transport of UDP-N acetylglucosamine into its Golgi lumen can be rescued by the wild-type SLC35A3 gene, but not by the mutated gene. These results provide the first demonstration of a genetic disorder associated with a defective SLC35A3 gene, and reveal a new mechanism for malformation of the vertebral column caused by abnormal nucleotide sugar transport into the Golgi apparatus. PMID- 16344556 TI - The evolutionary dynamics of alpha-satellite. AB - Alpha-satellite is a family of tandemly repeated sequences found at all normal human centromeres. In addition to its significance for understanding centromere function, alpha-satellite is also a model for concerted evolution, as alpha satellite repeats are more similar within a species than between species. There are two types of alpha-satellite in the human genome; while both are made up of approximately 171-bp monomers, they can be distinguished by whether monomers are arranged in extremely homogeneous higher-order, multimeric repeat units or exist as more divergent monomeric alpha-satellite that lacks any multimeric periodicity. In this study, as a model to examine the genomic and evolutionary relationships between these two types, we have focused on the chromosome 17 centromeric region that has reached both higher-order and monomeric alpha satellite in the human genome assembly. Monomeric and higher-order alpha satellites on chromosome 17 are phylogenetically distinct, consistent with a model in which higher-order evolved independently of monomeric alpha-satellite. Comparative analysis between human chromosome 17 and the orthologous chimpanzee chromosome indicates that monomeric alpha-satellite is evolving at approximately the same rate as the adjacent non-alpha-satellite DNA. However, higher-order alpha-satellite is less conserved, suggesting different evolutionary rates for the two types of alpha-satellite. PMID- 16344557 TI - Abundant novel transcriptional units and unconventional gene pairs on human chromosome 22. AB - Novel transcriptional units (TUs) are EST-supported transcribed features not corresponding to known genes. Unconventional gene pairs (UGPs) are pairs of genes and/or TUs sharing exon-to-exon cis-antisense overlaps or putative bidirectional promoters. Computational TU and UGP discovery followed by manual curation was performed in the entire published 34.9-Mb human chromosome 22 euchromatic sequence. Novel TUs (n = 517) were as abundant as known genes (n = 492) and typically did not have nonprimate DNA and protein homologies. One hundred seventy one (33%) of TUs, but only 13 (3%) of genes, both lacked nonprimate conservation and localized to gaps in the human-mouse BLASTZ alignment. Novel TUs were richer in exonic primate-specific interspersed repetitive elements (P = 0.001) and were more likely to rely on splice junctions provided by them, than were known genes: 19% of spliced TUs, versus 5% of spliced genes, had a splice site within a primate-specific repeat. Hence, novel TUs and known genes may represent different portions of the transcriptome. Two hundred nine (21%) of chromosome 22 transcripts participated in 77 cis-antisense and 42 promoter-sharing UGPs. Transcripts involved simultaneously in both UGP types were more common than was expected (P = 0.01). UGPs were nonrandomly distributed along the sequence: 89 (75%) clustered in distinct regions, the sum of which equaled 4.4 Mb (<13% of the chromosome). Eighty (67%) of the UGPs possessed significant locus structure differences between primates and rodents. Since some TUs may be functional noncoding transcripts and since the cis-regulatory potential of UGPs is well recognized, TUs and UGPs specific to the primate lineage may contribute to the genomic basis for primate-specific phenotypes. PMID- 16344558 TI - Systematic genome-wide annotation of spliceosomal proteins reveals differential gene family expansion. AB - Although more than 200 human spliceosomal and splicing-associated proteins are known, the evolution of the splicing machinery has not been studied extensively. The recent near-complete sequencing and annotation of distant vertebrate and chordate genomes provides the opportunity for an exhaustive comparative analysis of splicing factors across eukaryotes. We describe here our semiautomated computational pipeline to identify and annotate splicing factors in representative species of eukaryotes. We focused on protein families whose role in splicing is confirmed by experimental evidence. We visually inspected 1894 proteins and manually curated 224 of them. Our analysis shows a general conservation of the core spliceosomal proteins across the eukaryotic lineage, contrasting with selective expansions of protein families known to play a role in the regulation of splicing, most notably of SR proteins in metazoans and of heterogeneous nuclear ribonucleoproteins (hnRNP) in vertebrates. We also observed vertebrate-specific expansion of the CLK and SRPK kinases (which phosphorylate SR proteins), and the CUG-BP/CELF family of splicing regulators. Furthermore, we report several intronless genes amongst splicing proteins in mammals, suggesting that retrotransposition contributed to the complexity of the mammalian splicing apparatus. PMID- 16344559 TI - Molecular evolution and tempo of amplification of human LINE-1 retrotransposons since the origin of primates. AB - We investigated the evolution of the families of LINE-1 (L1) retrotransposons that have amplified in the human lineage since the origin of primates. We identified two phases in the evolution of L1. From approximately 70 million years ago (Mya) until approximately 40 Mya, three distinct L1 lineages were simultaneously active in the genome of ancestral primates. In contrast, during the last 40 million years (Myr), i.e., during the evolution of anthropoid primates, a single lineage of families has evolved and amplified. We found that novel (i.e., unrelated) regulatory regions (5'UTR) have been frequently recruited during the evolution of L1, whereas the two open-reading frames (ORF1 and ORF2) have remained relatively conserved. We found that L1 families coexisted and formed independently evolving L1 lineages only when they had different 5'UTRs. We propose that L1 families with different 5'UTR can coexist because they don't rely on the same host-encoded factors for their transcription and therefore do not compete with each other. The most prolific L1 families (families L1PA8 to L1PA3) amplified between 40 and 12 Mya. This period of high activity corresponds to an episode of adaptive evolution in a segment of ORF1. The correlation between the high activity of L1 families and adaptive evolution could result from the coevolution of L1 and a host-encoded repressor of L1 activity. PMID- 16344555 TI - The Oryza bacterial artificial chromosome library resource: construction and analysis of 12 deep-coverage large-insert BAC libraries that represent the 10 genome types of the genus Oryza. AB - Rice (Oryza sativa L.) is the most important food crop in the world and a model system for plant biology. With the completion of a finished genome sequence we must now functionally characterize the rice genome by a variety of methods, including comparative genomic analysis between cereal species and within the genus Oryza. Oryza contains two cultivated and 22 wild species that represent 10 distinct genome types. The wild species contain an essentially untapped reservoir of agriculturally important genes that must be harnessed if we are to maintain a safe and secure food supply for the 21st century. As a first step to functionally characterize the rice genome from a comparative standpoint, we report the construction and analysis of a comprehensive set of 12 BAC libraries that represent the 10 genome types of Oryza. To estimate the number of clones required to generate 10 genome equivalent BAC libraries we determined the genome sizes of nine of the 12 species using flow cytometry. Each library represents a minimum of 10 genome equivalents, has an average insert size range between 123 and 161 kb, an average organellar content of 0.4%-4.1% and nonrecombinant content between 0% and 5%. Genome coverage was estimated mathematically and empirically by hybridization and extensive contig and BAC end sequence analysis. A preliminary analysis of BAC end sequences of clones from these libraries indicated that LTR retrotransposons are the predominant class of repeat elements in Oryza and a roughly linear relationship of these elements with genome size was observed. PMID- 16344561 TI - Genome-wide mapping of DNase hypersensitive sites using massively parallel signature sequencing (MPSS). AB - A major goal in genomics is to understand how genes are regulated in different tissues, stages of development, diseases, and species. Mapping DNase I hypersensitive (HS) sites within nuclear chromatin is a powerful and well established method of identifying many different types of regulatory elements, but in the past it has been limited to analysis of single loci. We have recently described a protocol to generate a genome-wide library of DNase HS sites. Here, we report high-throughput analysis, using massively parallel signature sequencing (MPSS), of 230,000 tags from a DNase library generated from quiescent human CD4+ T cells. Of the tags that uniquely map to the genome, we identified 14,190 clusters of sequences that group within close proximity to each other. By using a real-time PCR strategy, we determined that the majority of these clusters represent valid DNase HS sites. Approximately 80% of these DNase HS sites uniquely map within one or more annotated regions of the genome believed to contain regulatory elements, including regions 2 kb upstream of genes, CpG islands, and highly conserved sequences. Most DNase HS sites identified in CD4+ T cells are also HS in CD8+ T cells, B cells, hepatocytes, human umbilical vein endothelial cells (HUVECs), and HeLa cells. However, approximately 10% of the DNase HS sites are lymphocyte specific, indicating that this procedure can identify gene regulatory elements that control cell type specificity. This strategy, which can be applied to any cell line or tissue, will enable a better understanding of how chromatin structure dictates cell function and fate. PMID- 16344562 TI - Transcription-mediated gene fusion in the human genome. AB - Transcription of a gene usually ends at a regulated termination point, preventing the RNA-polymerase from reading through the next gene. However, sporadic reports suggest that chimeric transcripts, formed by transcription of two consecutive genes into one RNA, can occur in human. The splicing and translation of such RNAs can lead to a new, fused protein, having domains from both original proteins. Here, we systematically identified over 200 cases of intergenic splicing in the human genome (involving 421 genes), and experimentally demonstrated that at least half of these fusions exist in human tissues. We showed that unique splicing patterns dominate the functional and regulatory nature of the resulting transcripts, and found intergenic distance bias in fused compared with nonfused genes. We demonstrate that the hundreds of fused genes we identified are only a subset of the actual number of fused genes in human. We describe a novel evolutionary mechanism where transcription-induced chimerism followed by retroposition results in a new, active fused gene. Finally, we provide evidence that transcription-induced chimerism can be a mechanism contributing to the evolution of protein complexes. PMID- 16344563 TI - Organization of the Caenorhabditis elegans small non-coding transcriptome: genomic features, biogenesis, and expression. AB - Recent evidence points to considerable transcription occurring in non-protein coding regions of eukaryote genomes. However, their lack of conservation and demonstrated function have created controversy over whether these transcripts are functional. Applying a novel cloning strategy, we have cloned 100 novel and 61 known or predicted Caenorhabditis elegans full-length ncRNAs. Studying the genomic environment and transcriptional characteristics have shown that two thirds of all ncRNAs, including many intronic snoRNAs, are independently transcribed under the control of ncRNA-specific upstream promoter elements. Furthermore, the transcription levels of at least 60% of the ncRNAs vary with developmental stages. We identified two new classes of ncRNAs, stem-bulge RNAs (sbRNAs) and snRNA-like RNAs (snlRNAs), both featuring distinct internal motifs, secondary structures, upstream elements, and high and developmentally variable expression. Most of the novel ncRNAs are conserved in Caenorhabditis briggsae, but only one homolog was found outside the nematodes. Preliminary estimates indicate that the C. elegans transcriptome contains approximately 2700 small non coding RNAs, potentially acting as regulatory elements in nematode development. PMID- 16344564 TI - Tandem chimerism as a means to increase protein complexity in the human genome. AB - The "one-gene, one-protein" rule, coined by Beadle and Tatum, has been fundamental to molecular biology. The rule implies that the genetic complexity of an organism depends essentially on its gene number. The discovery, however, that alternative gene splicing and transcription are widespread phenomena dramatically altered our understanding of the genetic complexity of higher eukaryotic organisms; in these, a limited number of genes may potentially encode a much larger number of proteins. Here we investigate yet another phenomenon that may contribute to generate additional protein diversity. Indeed, by relying on both computational and experimental analysis, we estimate that at least 4%-5% of the tandem gene pairs in the human genome can be eventually transcribed into a single RNA sequence encoding a putative chimeric protein. While the functional significance of most of these chimeric transcripts remains to be determined, we provide strong evidence that this phenomenon does not correspond to mere technical artifacts and that it is a common mechanism with the potential of generating hundreds of additional proteins in the human genome. PMID- 16344560 TI - Diversification of transcriptional modulation: large-scale identification and characterization of putative alternative promoters of human genes. AB - By analyzing 1,780,295 5'-end sequences of human full-length cDNAs derived from 164 kinds of oligo-cap cDNA libraries, we identified 269,774 independent positions of transcriptional start sites (TSSs) for 14,628 human RefSeq genes. These TSSs were clustered into 30,964 clusters that were separated from each other by more than 500 bp and thus are very likely to constitute mutually distinct alternative promoters. To our surprise, at least 7674 (52%) human RefSeq genes were subject to regulation by putative alternative promoters (PAPs). On average, there were 3.1 PAPs per gene, with the composition of one CpG-island containing promoter per 2.6 CpG-less promoters. In 17% of the PAP-containing loci, tissue-specific use of the PAPs was observed. The richest tissue sources of the tissue-specific PAPs were testis and brain. It was also intriguing that the PAP-containing promoters were enriched in the genes encoding signal transduction related proteins and were rarer in the genes encoding extracellular proteins, possibly reflecting the varied functional requirement for and the restricted expression of those categories of genes, respectively. The patterns of the first exons were highly diverse as well. On average, there were 7.7 different splicing types of first exons per locus partly produced by the PAPs, suggesting that a wide variety of transcripts can be achieved by this mechanism. Our findings suggest that use of alternate promoters and consequent alternative use of first exons should play a pivotal role in generating the complexity required for the highly elaborated molecular systems in humans. PMID- 16344565 TI - Experimental validation of the regulated expression of large numbers of non coding RNAs from the mouse genome. AB - Recent large-scale analyses of mainly full-length cDNA libraries generated from a variety of mouse tissues indicated that almost half of all representative cloned sequences did not contain an apparent protein-coding sequence, and were putatively derived from non-protein-coding RNA (ncRNA) genes. However, many of these clones were singletons and the majority were unspliced, raising the possibility that they may be derived from genomic DNA or unprocessed pre-mRNA contamination during library construction, or alternatively represent nonspecific "transcriptional noise." Here we show, using reverse transcriptase-dependent PCR, microarray, and Northern blot analyses, that many of these clones were derived from genuine transcripts of unknown function whose expression appears to be regulated. The ncRNA transcripts have larger exons and fewer introns than protein coding transcripts. Analysis of the genomic landscape around these sequences indicates that some cDNA clones were produced not from terminal poly(A) tracts but internal priming sites within longer transcripts, only a minority of which is encompassed by known genes. A significant proportion of these transcripts exhibit tissue-specific expression patterns, as well as dynamic changes in their expression in macrophages following lipopolysaccharide stimulation. Taken together, the data provide strong support for the conclusion that ncRNAs are an important, regulated component of the mammalian transcriptome. PMID- 16344566 TI - Comprehensive analysis of transcriptional promoter structure and function in 1% of the human genome. AB - Transcriptional promoters comprise one of many classes of eukaryotic transcriptional regulatory elements. Identification and characterization of these elements are vital to understanding the complex network of human gene regulation. Using full-length cDNA sequences to identify transcription start sites (TSS), we predicted more than 900 putative human transcriptional promoters in the ENCODE regions, representing a comprehensive sampling of promoters in 1% of the genome. We identified 387 fragments that function as promoters in at least one of 16 cell lines by measuring promoter activity in high-throughput transient transfection reporter assays. These positive functional results demonstrate widespread use of alternative promoters. We show a strong correlation between promoter activity and the corresponding endogenous RNA transcript levels, providing the first experimental quantitative estimate of promoter contribution to gene regulation. Finally, we identified functional regions within a randomly selected subset of 45 promoters using deletion analyses. These experiments showed that, on average, the sequence -300 to -50 bp of the TSS positively contributes to core promoter activity. Interestingly, putative negative elements were identified -1000 to -500 bp upstream of the TSS for 55% of genes tested. These data provide the largest and most comprehensive view of promoter function in the human genome. PMID- 16344567 TI - Development of a microscopic platform for real-time monitoring of biomolecular interactions. AB - We developed a new microscopic platform for the real-time analysis of molecular interactions by combining microbead-tagging techniques with total internal reflection fluorescent microscopy (TIRFM). The optical manipulation of probe microbeads, followed by photo immobilization on a solid surface, enabled us to generate arrays with extremely high density (>100 microbeads in a 25 microm x 25 microm area), and TIRFM made it possible to monitor the binding reactions of fluorescently labeled targets onto probe microbeads without removal of free targets. We demonstrated the high performance of this platform through analyses of interactions between antigen and antibody and between small compounds and proteins. Then, recombinant protein levels in total cellular lysates of Escherichia coli were quantified from the association kinetics using antibody immobilized microbead arrays, which served as a model for a protein-profiling array. Furthermore, in combination with in vitro synthesis-coupled protein labeling, we could kinematically analyze the interaction of nuclear factor kappaB (p50) with DNA. These results demonstrated that this platform enabled us to: (1) monitor binding processes of fluorescently labeled targets to multiple probes in real-time without removal of free targets, (2) determine concentrations of free targets only from the association kinetics at an early phase, and (3) greatly reduce the required volume of the target solution, in principle to subnanoliter, for molecular interaction analysis. The unique features of this microbead-based microarray system open the way to explore molecular interactions with a wide range of affinities in extremely small volumes of target solutions, such as extracts from single cells. PMID- 16344568 TI - Variation in crossing-over rates across chromosome 4 of Arabidopsis thaliana reveals the presence of meiotic recombination "hot spots". AB - Crossover (CO) is a key process for the accurate segregation of homologous chromosomes during the first meiotic division. In most eukaryotes, meiotic recombination is not homogeneous along the chromosomes, suggesting a tight control of the location of recombination events. We genotyped 71 single nucleotide polymorphisms (SNPs) covering the entire chromosome 4 of Arabidopsis thaliana on 702 F2 plants, representing 1404 meioses and allowing the detection of 1171 COs, to study CO localization in a higher plant. The genetic recombination rates varied along the chromosome from 0 cM/Mb near the centromere to 20 cM/Mb on the short arm next to the NOR region, with a chromosome average of 4.6 cM/Mb. Principal component analysis showed that CO rates negatively correlate with the G+C content (P = 3x10(-4)), in contrast to that reported in other eukaryotes. COs also significantly correlate with the density of single repeats and the CpG ratio, but not with genes, pseudogenes, transposable elements, or dispersed repeats. Chromosome 4 has, on average, 1.6 COs per meiosis, and these COs are subjected to interference. A detailed analysis of several regions having high CO rates revealed "hot spots" of meiotic recombination contained in small fragments of a few kilobases. Both the intensity and the density of these hot spots explain the variation of CO rates along the chromosome. PMID- 16344569 TI - Mucosal prolapse in the pathogenesis of Peutz-Jeghers polyposis. AB - Germline mutations in LKB1 cause the rare cancer prone disorder Peutz-Jeghers syndrome (PJS). Gastrointestinal hamartomatous polyps constitute the major phenotypic trait in PJS. Hamartomatous polyps arising in PJS patients are generally considered to lack premalignant potential although rare neoplastic changes in these polyps and an increased gastrointestinal cancer risk in PJS are well documented. These conflicting observations are resolved in the current hypothesis by providing a unifying explanation for these contrasting features of PJS polyposis. We postulate that a genetic predisposition to epithelial prolapse underlies the formation of the polyps associated with PJS. Conventional sporadic adenomas arising in PJS patients will similarly show mucosal prolapse and carry the associated histological features. PMID- 16344570 TI - Is thiopurine therapy in ulcerative colitis as effective as in Crohn's disease? PMID- 16344571 TI - Survival in colorectal cancer: impact of body mass and exercise. PMID- 16344572 TI - Combined PET/CT colonography: is this the way forward? PMID- 16344573 TI - Pancreatic stellate cells: new kids become mature. PMID- 16344574 TI - Rectal bleeding and abdominal mass. PMID- 16344575 TI - A rare cause of diuretic refractory ascites. PMID- 16344576 TI - Gastrointestinal bleeding: don't overlook the role of the pancreas. PMID- 16344577 TI - Hypervascular pancreatic mass. PMID- 16344579 TI - DNA test for hypolactasia premature. PMID- 16344580 TI - IL-1 gene cluster and TNFA-307 polymorphisms in the risk of perforated duodenal ulcer. PMID- 16344578 TI - Steatosis in chronic hepatitis C: why does it really matter? AB - Hepatic steatosis is a common histological feature of chronic hepatitis C. Various factors are associated with hepatic steatosis, including obesity, high alcohol consumption, diabetes type II, and hyperlipidaemia. These factors may contribute to steatosis in patients with chronic hepatitis C. In humans, hepatitis C virus (HCV) genotype 3 is more commonly associated with steatosis. In vitro studies and the transgenic mouse model have suggested that the HCV core protein (genotype 1) can induce lipid accumulation within hepatocytes. However, what is the relevance of steatosis in chronic hepatitis C? It seems that in certain populations, steatosis may be associated with fibrosis progression and this may be genotype specific. The mechanisms underlying this association are unknown; neither is it clear whether this holds true for all patients or only a subgroup. Indeed, after antiviral treatment, virus related steatosis disappears whereas the host associated steatosis remains unaffected. This review describes and discusses the basic and clinical aspects of the relationship between steatosis and progression of fibrosis, and response to treatment in patients with chronic hepatitis C. PMID- 16344581 TI - "Cannabis hyperemesis" causation questioned. PMID- 16344582 TI - Cryptic gluten intolerance in type 1 diabetes: identifying suitable candidates for a gluten free diet. PMID- 16344583 TI - Acquired factor V inhibitor associated with valproic acid use in a cirrhotic patient. PMID- 16344584 TI - Future use of the Glasgow alcoholic hepatitis score. PMID- 16344585 TI - Acylated ghrelin stimulates food intake in the fed and fasted states but desacylated ghrelin has no effect. PMID- 16344586 TI - A proof of concept study establishing Necator americanus in Crohn's patients and reservoir donors. PMID- 16344587 TI - Strong BCL10 nuclear expression identifies gastric MALT lymphomas that do not respond to H pylori eradication. PMID- 16344593 TI - Molecular pathogenesis of alcohol-induced hepatic fibrosis. AB - Alcohol abuse is a main cause of liver fibrosis and cirrhosis in the western world. Although the major mechanisms of fibrogenesis are independent of the origin of liver injury, alcoholic liver fibrosis features distinctive characteristics, including the pronounced inflammatory response of immune cells due to elevated gut-derived endotoxin plasma levels, increased formation of reactive oxygen species (ROS), ethanol-induced pericentral hepatic hypoxia or formation of cell-toxic and pro-fibrogenic ethanol metabolites (e.g., acetaldehyde or lipid oxidation products). These factors are together responsible for increased hepatocellular cell death and activation of hepatic stellate cells (HSCs), the key cell type of liver fibrogenesis. To date, removing the causative agent is the most effective intervention to prevent the manifestation of liver cirrhosis. A novel experimental approach in fibrosis therapy is the selective induction of cell death in HSCs. Substances such as gliotoxin, anandamide or antibody against tissue inhibitor of metalloproteinase (TIMP)-1 can selectively induce cell death in activated HSCs. These new results in basic science are encouraging for the search of new antifibrotic treatment. PMID- 16344588 TI - Interferon-beta plus ribavirin for patients with hepatitis C virus genotype 1: a randomised pilot trial. PMID- 16344594 TI - Oxidative stress and signal transduction pathways in alcoholic liver disease. AB - Ethanol is linked to several pathologies like alcohol liver injury, neurotoxicity, cardiomyopathy, fetal alcoholic syndrome or cancer. It is generally accepted that oxidative stress plays a central role in their pathogenesis. After chronic and excessive consumption, alcohol may accelerate oxidative mechanisms both directly via increased production of reactive oxygen species and indirectly by impairing protective mechanisms against them. Ethanol, its metabolites arising during its metabolic degradation as well as novel compounds formed via ethanol induced oxidative stress, especially during the action of the ethanol inducible microsomal cytochrome CYP2E1, may apart from direct damage to biological structures affect signal transduction pathways thus modulating and potentiating damage. Alteration of the redox status of cells following chronic ethanol misuse may have profound effects on cellular function and viability and lead to cell death and tissue damage. These changes linked to pathologic processes in the organism, are related to alteration of intracellular signaling pathways associated with protein kinases and transcription factor activation. Mainly mitogen activated protein kinase (MAPK) family, transcription factors-nuclear factor kappaB (NF-kappaB) and activating protein 1 (AP-1) are involved in the deterioration of cells and organs. The response is cell-type specific and depends on the dose of ethanol. Oxido-reduction balance, regulatory disturbances and signal transduction cascades responsible for alcoholic damage have been partially described, nevertheless, further studies are required to allow future novel diagnostic and therapeutical strategies. We are only at the beginning ... PMID- 16344595 TI - Sesamin ingestion regulates the transcription levels of hepatic metabolizing enzymes for alcohol and lipids in rats. AB - BACKGROUND: Sesamin, a major lignan in sesame seeds, has multiple functions such as stimulation effect of ethanol metabolism in mice and human, and prevention of ethanol-induced fatty liver in rats. However, the mechanism has not been clarified yet. METHODS: The changes of gene expression were investigated in rats given 250 mg/kg of sesamin (sesamin rats) or vehicle (control rats) for three days by using a DNA microarray analysis. At 4 hr after the final ingestion, the profiles of gene expression in rat livers were compared. RESULTS: The analysis showed that 38 transcripts were up-regulated with a significant change of more than two-fold and eight transcripts were down-regulated with a significant change to less than half in the livers of sesamin rats versus control rats. The gene expression levels of the early stage enzymes of beta-oxidation including long chain acyl-CoA synthetase, very long-chain acyl-CoA synthetase and carnitine palmitoyltransferase were not changed, however, those of the late stage enzymes of beta-oxidation including trifunctional enzyme in mitochondria, and acyl-CoA oxidase, bifunctional enzyme and 3-ketoacyl-CoA thiolase in peroxisomes, were significantly increased by sesamin ingestion. Also, in sesamin rats, the gene expression of aldehyde dehydrogenase was increased about three-fold, whereas alcohol dehydrogenase, liver catalase and CYP2E1 were not changed. Changes in the gene expression of alcohol- and aldehyde-metabolizing enzymes observed in a DNA microarray were also confirmed by a real-time PCR method. CONCLUSIONS: These results suggested that sesamin ingestion regulated the transcription levels of hepatic metabolizing enzymes for alcohol and lipids. PMID- 16344596 TI - Anti-TGF-beta strategies for the treatment of chronic liver disease. AB - Permanent alcohol abuse may lead to chronic liver injury with deleterious sequelae such as liver cirrhosis and hepatocellular carcinoma. Mechanisms of fibrogenesis encompass recruitment of inflammatory cells at the site of injury and cytokine mediated activation of hepatic stellate cells (HSC) with accumulation of interstitial collagens. HSC transdifferentiation and accompanying apoptosis result in destruction of liver architecture and are therefore key steps of disease progression. TGF-beta represents the main profibrogenic cytokine in liver fibrosis and other fibroproliferative disorders by inducing extracellular matrix deposition as part of the wound healing response. In parallel, TGF-beta triggers hepatocytes that are strongly responsive for this cytokine, to undergo apoptosis, thereby providing space for HSC proliferation and generation of a collagenous matrix. Anti TGF-beta approaches were established and successfully utilized for the treatment of experimental fibrogenesis. Dominant negative TGF beta receptors (TbetaR), generated by fusing the Fc domain of human IgG and the N terminal (extracellular) fragment of TbetaRII (Fc:TbetaRII) were applied to suppress fibrosis. Similarly TGF-beta binding proteins like decorin, antagonistic cytokines such as bone morphogenetic protein-7, hepatocyte growth factor, IL-10, or IFN-gamma were as efficient as camostat mesilate, a protease inhibitor that possibly abrogated proteolytic activation of TGF-beta. Further, our group recently overexpressed Smad7 in bile duct ligation induced liver fibrosis and achieved efficient inhibition of intracellular TGF-beta signaling, thereby counteracting profibrogenic effects in cultured HSC and in vivo. A direct link between the effect of alcohol and TGF-beta exists through reactive oxygen species that are generated in liver cells by alcohol metabolism and represent activators of TGF-beta signaling. Thus, soluble TbetaRII expression reduced experimental fibrogenesis in vitro and in vivo partially by decreasing intracellular ROS and inhibiting NADH oxidase. Approaches that specifically target profibrogenic TGF beta signaling are promising to treat alcoholic liver disease in the future. However, to ensure safety for the patients to be treated, approaches with strong specificity need to be established. Therefore, it is essential to delineate the profibrogenic actions of TGF-beta and the influence of alcohol abuse in molecular detail. PMID- 16344597 TI - Adipogenic phenotype of hepatic stellate cells. AB - Transdifferentiation of hepatic stellate cells (HSC) constitutes a major cellular event in the genesis of alcoholic liver fibrosis and cirrhosis and molecular mechanisms underlying this process is incompletely understood. Our laboratory proposed several years ago that HSC quiescence requires the transcriptional program known to be integral to preadipocyte to adipocyte differentiation. In support of the hypothesis, our research demonstrates the expression of adipogenic transcription factors (C/EBPs, PPARgamma, SREBP-1c, LXRalpha) and adipocyte specific genes (adipsin, resistin) are high in quiescent HSC and depleted in activated HSC. Three gain-of-function approaches have been taken to test this notion: the treatment of activated HSC with the adipocyte differentiation cocktail; ectopic expression of PPARgamma or SREBP-1c. All three treatments coordinately upregulate a panel of putative adipogenic transcription factors and cause morphologic and biochemical reversal of activated HSC to quiescent cells. These findings establish a new conceptual framework for the treatment of liver fibrosis and propose an intriguing notion concerning the plasticity of HSC. PMID- 16344598 TI - Carbon monoxide as a guardian against hepatobiliary dysfunction. AB - Carbon monoxide (CO) generated through the reaction of heme oxygenase (HO) has attracted great interest in regulation of hepatobiliary homeostasis. The gas generated by HO-2 in the hepatic parenchyma can modestly activate soluble guanylate cyclase (sGC) expressed in hepatic stellate cells in a paracrine manner and thereby constitutively relax sinusoids. Kupffer cells express HO-1, the inducible isozyme, even under normal unstimulated conditions and constitutes approximately 30% of the total HO activity in this organ. Upon exposure to a variety of stressors such as cytokines, endotoxin, hypoxia and oxidative stress, the liver induces HO-1 and over-produces CO. The stress-inducible CO has been shown to guarantee ample blood supply during detoxification of heme and thus to play a protective role in the liver. However, molecular mechanisms by which CO serves as a protectant for hepatocytes, the cells expressing little sGC, remain to be solved. Previous observation suggested that CO modulates intracellular calcium mobilization through inhibiting cytochrome P-450 activities and thereby maintain stroke volume of bile canalicular contraction in cultured hepatocytes. CO also stimulates mrp2-dependent excretion of bilirubin-IXalpha and helps heme catabolism. Although a direct molecular target responsible for the latter event remains unknown, such properties of CO could support xenobiotic metabolism through its actions on sinusoidal hemodynamics and hepatobiliary systems. PMID- 16344599 TI - Modulation of non-alcoholic steatohepatitis by pattern recognition receptors in mice: the role of toll-like receptors 2 and 4. AB - Toll-like receptors (TLR) recognize pathogen-derived molecules and induce downstream activation of inflammatory pathways. Fatty liver has been shown to result in increased sensitivity to lipopolysaccharide (LPS), a TLR4 ligand. In this study, we investigated the roles of TLR2 and TLR4 in liver damage and on cytokine induction in a methionine-choline deficient (MCD) diet-induced model of nonalcoholic steatohepatitis. We found that mice with nonalcoholic fatty liver had increased liver injury and inflammatory cytokine induction after challenge with a TLR4 but not with a TLR2 ligand. TLR2 deficient mice were not protected against the development of steatohepatitis after MCD diet feeding. On the contrary, TLR2 mice had significantly higher levels of serum ALT and greater TNF alpha levels after LPS challenge suggesting increased liver injury. This was associated with reduced production of IL-6, a cytokine with hepatoprotective effects in fatty liver. Increased liver injury in the MCD diet-fed TLR2 mice was associated with reduced baseline and LPS-induced NF-kB and PPRE binding compared to MCS controls. These results demonstrate that TLR2 deficiency results in increased liver injury in association with nonalcoholic steatohepatitis and may suggest a protective role for TLR2-mediated signals in liver injury. PMID- 16344600 TI - Ethanol-induced liver injury: potential roles for egr-1. AB - Chronic ethanol-induced liver injury follows a typical progression from its earliest stage of steatosis to more advanced injury, characterized by the development of inflammation, hepatocyte necrosis/apoptosis, fibrosis and finally cirrhosis. Kupffer cells, the resident macrophage in the liver, play a critical role in the progression of liver injury. Increased exposure of Kupffer cells to lipopolysaccharide (LPS) during chronic ethanol exposure leads to the production of a number of inflammatory mediators, including tumor necrosis factor alpha (TNF alpha). Recent evidence indicates that in addition to increased exposure to LPS, Kupffer cells also develop an enhanced sensitivity to LPS after chronic ethanol feeding. We have recently identified early growth response-1 (Egr-1), an immediate-early gene transcription factor, as an important contributor to increased LPS-stimulated TNF-alpha secretion by Kupffer cells after chronic ethanol exposure. In other models of tissue injury, such as ischemia-reperfusion in the lung, Egr-1 acts as a coordinator of the complex response to stress. Here we review the literature regarding the role of EGR-1 in regulation of a number of genes implicated in each of the stages of chronic ethanol-induced liver injury. In addition to the critical role of Egr-1 in generating maximal LPS-stimulated TNF-alpha expression, Egr-1 also controls the expression of a number of inflammatory mediators, including intercellular adhesion molecule (ICAM)-1, monocyte chemotactic protein (MCP)-1 and macrophage inflammatory protein (MIP)-2, as well as genes contributing to fibrosis, such as transforming growth factor (TFG)-beta1, platelet-derived growth factor PDGF-A chain and fibroblast growth factor (FGF). Understanding the contribution of Egr-1 to the expression of genes involved in the development of chronic ethanol-induced liver injury may lead to the development of improved therapies designed to prevent and/or reverse alcohol induced liver injury. PMID- 16344601 TI - The pathogenesis and significance of the urinary alcohol cycle in rats fed ethanol intragastrically. AB - The blood alcohol level cycle (BALC) of the intragastric tube feeding model first described by Tsukamoto et al., has three separate essential mechanistic components. The first is the requirement for an intact functioning thyroid. The evidence for this is that propylthiouracil or severance of the pituitary stalk completely prevents the cycle. What happens instead of the cycle is that the blood alcohol level rises to a lethal level when ethanol is given continuously at a dose of 11 g/kg/day by stomach tube. When excess thyroid hormone is given orally it markedly attenuates the cycle because it interferes with the changes in the level of thyroid hormone during the cycle. The second component is norepinephrine. Catecholamines are markedly elevated at the peaks of the cycle. Both propranolol and phenoxybenzamine, which are beta- and alpha-blockers, prevent the cycle. Also, when catecholamines are fed in excess in the form of ephedrine, the cycle is eliminated. The third element essential to the cycle is the generation of NAD to support the oxidation of alcohol by alcohol dehydrogenase. When complex I (NADH dehydrogenase) of the mitochondrial electron transport chain is inhibited by feeding rotenone, the cycle is totally eliminated and blood alcohol levels remain constant at 200 mg/%. Thus NADH increases and NAD decreases at the peak of the cycle. Without the fluxuation of NAD, ADH activity cannot fluctuate during the cycle and the cycle is prevented. The significance of the BALC in the understanding of alcohol liver disease pathogenesis is that there's a marked difference in the gene expression and liver toxicity when the peaks and troughs of the cycle are compared. The expression of 1000+ genes is either two-fold up or down regulated as determined by microarray analysis. At the peaks there is increased liver pathology, especially inflammatory changes in the liver associated with an increase of iNOS expression. The genes responsive to hypoxia inducible factor 1alpha (HIF1alpha) regulation are increased including the expression of erythropoietin, adrenomedullin and adrenergic receptor alpha 1a and d. The expression of prolyl hydroxylase, which destabilizes HIF1alpha, increases when the BAL drops to low levels during the cycle. The level of oxygen, as measured on the surface of the liver, is decreased at the peaks, compared to control livers. The NADH/NAD ratio is markedly increased and ATP levels are markedly decreased at the BAL peaks. Also, endotoxin in the blood is very high at the peaks and very low at the troughs. When the blood alcohol levels fall during the cycle, there is an increase in ALT, suggesting that reoxygenation from the hypoxic state at the peaks causes an ischemic reperfusion injury-like lesion in the liver. At this time there is also an increase in expression of many important enzymes such as manganese SOD. Genes such as c-fos and CTGF are increased in expression. These contrasting findings at the peaks and troughs indicate that the blood alcohol levels, which fluctuate up and down, change the gene expression and the pathology of the liver. PMID- 16344602 TI - Ceramide, tumor necrosis factor and alcohol-induced liver disease. AB - BACKGROUND: The pathogenesis of alcohol-induced liver disease (ALD) is incompletely known. One of the key processes mediating the progression of ALD involved the overproduction of tumor necrosis factor (TNF) and the susceptibility of hepatocytes to TNF-induced apoptosis by alcohol intake. METHODS: Analyze the apoptotic signaling of TNF resulting in the targeting and subsequent recruitment of mitochondria to death pathways. RESULTS: Studies in experimental animal models of the disease have provided evidence for the role of ceramide generated from acidic sphingomyelinase in the apoptotic signaling of TNF through recruitment of mitochondria. The mitochondrial pool of glutathione (mGSH) is a vital line of defense against oxidative stress by precluding the accumulation peroxides generated endogenously within mitochondria and as a cofactor of mitochondrial antioxidant enzymes. The depletion of mGSH by alcohol has been described to determine the susceptibility of hepatocytes to TNF-mediated cell death. CONCLUSIONS: The level of mGSH determines the fate of hepatocytes to acidic sphingomyelinase activation by TNF and hence strategies aimed to replenish mGSH or to antagonize the generation of ceramide from acidic sphingomyelinase may be of therapeutic value for ALD. PMID- 16344603 TI - Glycine as a therapeutic immuno-nutrient for alcoholic liver disease. AB - Activation of Kupffer cells by gut-derived endotoxin is an important factor in ethanol hepatotoxicity. Further, it was shown that ethanol modulates both the expression and activity of several intracellular signaling molecules and transcription factors in Kupffer cells and chronic ethanol treatment enhances Kupffer cell sensitivity to endotoxin. These findings suggest that inhibition of Kupffer cell activation is effective for clinical application in alcoholic hepatitis. Recently, accumulating lines of evidence suggest a possibility that glycine is useful as an immuno-modulating amino acid. It has been shown that a diet containing glycine improved survival in endotoxin shock by preventing Kupffer cell activation. Glycine most likely prevents the LPS-induced elevation of intracellular Ca concentration in Kupffer cells, thereby minimizing LPS receptor signaling and cytokine production. Indeed, glycine prevents alcohol induced liver injury in a long-term enteral ethanol feeding rats (Tsukamoto French) by decreasing production of TNF-alpha in the liver. Moreover, glycine is protective against apoptosis of sinusoidal endothelial cells (SECs) that is one of the initial events in the development of liver injury. On the other hand, epidemiologic data have identified chronic alcohol consumption as a significant risk factor for carcinogenesis. Interestingly, glycine inhibits growth of tumor in vivo most likely because of the inhibition of angiogenesis. It was shown that the inhibitory effect of glycine on growth and migration of endothelial cells is due to activation of a glycine-gated Cl channel. It is hypothesized that the opening of this anion channel hyperpolarizes the cell membrane, blocks influx of Ca through voltage-dependent Ca channel, thereby blunting growth factor-mediated signaling. Therefore, glycine can be used not only for treatment of alcoholic hepatitis, but also for chemoprevention and treatment of hepatocellular carcinoma in alcoholic cirrhosis. Taken together, it is concluded that glycine is a potent therapeutic immuno-nutrient for various kinds of chronic liver diseases including alcoholic liver disease (ALD). PMID- 16344604 TI - Activation of the innate immune system and alcoholic liver disease: effects of ethanol per se or enhanced intestinal translocation of bacterial toxins induced by ethanol? AB - It is generally accepted that activation of the innate immune system and increased release of pro-inflammatory cytokines and other mediators plays an important role in the development of alcoholic liver disease (ALD). The mechanisms involved in the ethanol-induced activation of monocytes/macrophages (including Kupffer cells) are however, still a matter of debate. The brief review will summarize the published data from the literature on the two main pathomechanisms discussed until now: I) Gut-derived bacterial toxins, specially endotoxin; and II) metabolic changes induced by alcohol oxidation (independent of mechanism I). For pathomechanism I, clear evidence has been published from numerous groups: Alcohol induces mucosal injury in the upper gastrointestinal tract and leads to marked increase in the permeability of the gut mucosa to macromolecules such as endotoxin. The resulting endotoxemia then leads to activation of Kupffer cells and other macrophages. The increased release of pro inflammatory mediators (e.g., TNF-alpha, Il-1, reacting oxygen species) and infiltration of other inflammatory cells (e.g., neutrophils) finally causes liver damage. Regarding the second pathomechanism it has repeatedly been argued that the metabolic alterations which are induced by chronic administration of ethanol to rats or mice might increase the sensitivity of monocytes/macrophages to secrete TNF-alpha and other pro-inflammatory mediators thereby increasing the susceptibility to ethanol-induced liver injury. However, in all feeding experiments the effect of ethanol on intestinal permeability and enhanced translocation of bacterial toxins (endotoxin) is likely to occur (or at least cannot be excluded). The latter holds true also for experiments using isolated macrophages/Kupffer cells from ethanol fed animals. Therefore, to clarify whether or not alterations related to ethanol metabolism ("direct" effects of ethanol) contribute to the activation of the innate immune system studies using germ-free animals are needed to exclude the "indirect" effect of ethanol via gut-derived bacterial toxins. PMID- 16344605 TI - Relation of endotoxin, endotoxin binding proteins and macrophages to severe alcoholic liver injury and multiple organ failure. AB - Our retrospective analysis of 105 patients with alcoholic liver injury confirmed that patients with severe alcoholic hepatitis (SAH) showed severe hyperbilirubinemia, reduced hepatic biosynthetic capacity, and marked acute inflammatory reactions, and developed multiple organ failure (MOF). Multivariate analysis using the Cox proportional hazards model showed serum C-reactive protein and DIC as significant independent prognostic factors among SAH, LC+AH, and AH groups. Improved assay showed an increase of plasma endotoxin with the progression of alcoholic liver injury. In most survivors, plasma Et levels decreased in the recovery phase. Serum interleukin (IL)-6 and IL-8 levels in the acute phase were high in patients with AH and LC+AH, especially in non-survivors and in patients with SAH. In the recovery phase, these cytokine levels in survivors tended to decrease, but in non-survivors, IL-6 remained high, and IL-8 further increased. Serum levels of HDL and albumin, which are protective against endotoxicity by inhibiting endotoxin uptake and TNF production by macrophages, were decreased with the progression of alcoholic liver injury. Animal experiments supported that the increase in endotoxin-binding capacity of HDL and albumin may serve as a protective mechanism against endotoxin in chronic ethanol-loaded rats and that an addition of high-dose ethanol to these rats may lead to impaired binding and inactivation of endotoxin. Lipopolysaccharide-binding protein (LBP) which enhances endotoxin uptake and TNF production by macrophages, was generally increased in patients with alcoholic liver injury. This imbalance among endotoxin binding proteins in the blood may induce overproduction of cytokines by macrophages in patients with severe alcoholic liver injury. Our animal experiments further revealed that an additional administration of a high-dose ethanol to chronic alcohol-fed rats led to decrease of endotoxin clearance, increased extrahepatic accumulation of endotoxin and elevation of plasma TNF. The splenic macrophages and pulmonary alveolar macrophages are demonstrated to be important for endotoxin uptake, and excessive production of TNF in rats given large amounts of alcohol. An in vitro culture experiment in the presence of rat LBP suggested a role of these macrophages in excessive production of TNF-alpha. When the functions of various macrophages were compared in rats given alcohol, maximum TNF-alpha secretion was noted in alveolar macrophages, In conclusion, endotoxemia and its effects on extrahepatic macrophages may play key roles in the progression of severe alcoholic liver injury and MOF. PMID- 16344606 TI - Dysregulated cytokine metabolism, altered hepatic methionine metabolism and proteasome dysfunction in alcoholic liver disease. AB - Alcoholic liver disease (ALD) remains an important complication and cause of morbidity and mortality from alcohol abuse. Major developments in our understanding of the mechanisms of ALD over the past decade are now being translated into new forms of therapy for this disease process which currently has no FDA approved treatment. Cytokines are low molecular weight mediators of cellular communication, and the pro-inflammatory cytokine tumor necrosis factor (TNF) has been shown to play a pivotal role in the development of experimental ALD. Similarly, TNF levels are elevated in the serum of alcoholic hepatitis patients. Abnormal methionine metabolism is well documented in patients with ALD, with patients having elevated serum methionine levels, but low S adenosylmethionine levels in the liver. On the other hand, S-adenosylhomocysteine and homocysteine levels are elevated in ALD. Recent studies have documented potential interactions between homocysteine and S-adenosylhomocysteine with TNF in the development of ALD. Altered proteasome function also is now well documented in ALD, and decreased proteasome function can cause hepatocyte apoptosis. Recently it has been shown that decreased proteasome function can also act synergistically to enhance TNF hepatotoxicity. Hepatocytes dying of proteasome dysfunction release pro-inflammatory cytokines such as Interleukin-8 to cause sustained inflammation. This article reviews the interactions of cytokines, altered methionine metabolism, and proteasome dysfunction in the development of ALD. PMID- 16344607 TI - Iron accumulation in alcoholic liver diseases. AB - Increased hepatic iron is one of the important key factors which contribute alcohol toxicity of liver due to the production of reactive oxygen species. In patients with alcoholic liver diseases (ALD), liver iron is increased and the resulted lipid metabolite 4-hydroxynonenal-protein adduct was also increased. In general, iron is deposited in both parenchymal cells and and Kupffer cells in ALD. However, in patients with mild ALD, the parenchymal iron deposition is dominant rather than reticuloendothelial iron deposition, while the latter iron deposition is domimant in severe ALD, possibly due to endotoxemia and overproduction of inflammatory cytokines. We speculated that a parenchymal iron deposition in mild ALD is an important factor to trigger hepatocytes injury by ethanol, and the possible cause of parencynal iron deposition may be an increase of cellular iron uptake via serum transferrin in hepatocytes after ethanol exposure. By immuno-histochemical study of biopsied liver samples, the expression of transferrin receptor 1 (TfR1), which mediates cellular iron uptake by serum transferrin was increased. This increase of TfR1 by ethanol is confirmed by in vitro experiment using HepG2 cells and primary rat hepatocytes culture. Fe labeled transferrin incorporation (but not transferrin non-bound iron (NTBI)) into the cells is also increased, suggesting that the increased TfR1 is functional. The increase of TfR1 expression is partially due to the increased activity of iron regulatory protein (IRP) by oxidative stress of ethanol metabolism. Thus, the post-transcriptional regulation of iron uptake by ethanol is involved in the hepatocyte iron accumulation. Another possibility is an increase of intestinal iron absorption. Our recent finding regarding the increase of pro-hepcidin serum in alcoholic patients with high serum ferritin support this assumption. PMID- 16344608 TI - Analgesic selection in the management of chronic pain: linking mechanisms & evidence-based research to clinical practice. PMID- 16344609 TI - Management of neuropathic pain: translating mechanistic advances and evidence based research into clinical practice. AB - The concept of rational polypharmacy is now well established in the field of pain management. This concept has evolved in concert with progress in understanding the pathophysiologic mechanisms of pain diseases and disorders and how medications affect these processes. Other clinical factors must be considered in formulating the pain management strategy most likely to succeed in both controlling pain and improving function in a given patient. This article will review how pain diagnosis, pain mechanisms, pain phenomenology, medication efficacy, and risk profile influence medication selection in pain medicine practice, with a selective focus on the treatment of neuropathic pain. In addition, the role of psychosocial factors as they affect pain management will be discussed. PMID- 16344610 TI - Using pain quality assessment measures for selecting analgesic agents. AB - Although those in the field of pain management have long recognized the multidimensional character of pain, global pain intensity remains the most frequently assessed domain measured in clinical trials of pain treatments. However, a number of specific pain qualities have been shown to be associated with neuropathic versus nociceptive pain and, as such, can be used for diagnostic purposes. In addition, preliminary evidence suggests that pain quality measures may be helpful for measuring outcomes in clinical trials. A number of such measures are currently available, each with its strengths and weaknesses. These measures are reviewed, and potential future uses of these tools are discussed. PMID- 16344611 TI - Bibliography. Current world literature. Systemic disorders with rheumatic manifestations. PMID- 16344612 TI - Systemic vasculitis: state of the art and emerging concepts. PMID- 16344613 TI - New approaches in the treatment of Adamantiades-Behcet's disease. AB - PURPOSE OF REVIEW: To update clinicians on the recent advances in the treatment of Adamantiades-Behcet's disease. RECENT FINDINGS: Interferon-alpha-2a and infliximab have proved able to induce prompt remission in the vast majority of Adamantiades-Behcet's patients with DMARD-resistant uveoretinitis. Efficacy of interferon-alpha-2a has also been reported for mucocutaneous lesions, arthritis, and (more anecdotally) for neuro-Behcet, while results from small case series suggest that infliximab is beneficial for mucocutaneous lesions and (more anecdotally) for arthritis and gastro-intestinal manifestations. Two cases of neuro-Behcet treated with infliximab showed a complete resolution. Finally, in a randomized controlled trial of patients with mucocutaneous, arthritic manifestations, or both, etanercept effectively suppressed mucocutaneous lesions.A different approach is tolerization by oral administration of the 336 351 peptide of the human heat shock protein 60 (thought to have a pathogenic role in Adamantiades-Behcet's disease-associated uveitis), linked to recombinant cholera B-toxin B-subunit. Preliminary results have shown that tolerization is safe and effective in preventing relapses of uveitis. SUMMARY: Biologic agents have proved effective in patients resistant to conventional treatment. However, disease subsets characterized by severe morbidity and mortality such as vasculo Behcet and neuro-Behcet still pose major therapeutic challenges. Further studies are needed to devise better treatment strategies for severe Adamantiades-Behcet's disease. PMID- 16344614 TI - Large-vessel involvement in giant cell arteritis. AB - PURPOSE OF REVIEW: Large-vessel involvement in giant cell arteritis occurs in over a quarter of patients with this disease. Stenosis of the primary and secondary branches of the aorta may cause claudication and tissue gangrene, whereas aortitis may lead to aneurysm formation and dissection, often many years after the initial diagnosis. RECENT FINDINGS: Subsets of giant cell arteritis are probably caused by variations in the pathobiology of the disease. Radiographic imaging with some form of angiography is essential to reach a proper diagnosis. Although survival in giant cell arteritis is generally good, subsets of patients with aneurysm formation have a markedly diminished life expectancy. Glucocorticosteroids continue to be the mainstay of treatment for giant cell arteritis and its complications. SUMMARY: Advances in diagnostic techniques including proteomic and genomic approaches should improve our understanding of the pathogenesis as well as the assessment of disease activity and extent. We provide a suggested algorithm for the evaluation of patients with large-vessel disease in giant cell arteritis. PMID- 16344615 TI - Thromboangiitis obliterans (Buerger's disease). AB - PURPOSE OF REVIEW: Thromboangiitis obliterans is a nonatherosclerotic segmental inflammatory disease that affects the small and medium-sized arteries and veins in the upper and lower extremities. This review will help to familiarize physicians with this vasculitis that is completely different from every other type of vasculitis encountered. RECENT FINDINGS: While tobacco is central to the initiation and continuance of Buerger's disease activity, two interesting pathophysiologic observations have been made. There is endothelial dysfunction in arteries not yet clinically or angiographically involved in thromboangiitis obliterans. There are elevated levels of anti-endothelial cell antibodies and measurement of these antibody titers may be useful in following disease activity in Buerger's disease patients. The only therapy clearly shown to prevent amputation is the complete abstinence of tobacco. There is exciting work under way on the use of selective cannabinoid receptor antagonists to help patients stop smoking. In addition, preliminary results on use of therapeutic angiogenesis in patients with Buerger's disease has demonstrated excellent collateral blood vessels formation and clinical improvement. SUMMARY: The difficulty in studying rare diseases such as thromboangiitis obliterans is that there are no significant research dollars available and even the most active centers only see a few patients per year. Therefore, there has been little progress in understanding the pathogenesis of the disease. There are new therapeutic modalities that help patients with this disease, however, and patients can be assured that if they are able to discontinue tobacco use completely, amputation will not occur if critical limb ischemia is not already present. PMID- 16344616 TI - Advances in the therapy of Wegener's granulomatosis. AB - PURPOSE OF REVIEW: In the past, recommendations for the treatment of Wegener's granulomatosis were primarily based on findings reported from open-label clinical trials. Results from several randomized controlled trials in patients with Wegener's granulomatosis and other antineutrophil cytoplasm antibody-associated vasculitides have recently been reported that have a great impact on patient care. RECENT FINDINGS: In view of the considerable toxicity of cyclophosphamide, strategies to limit exposure to it have recently been evaluated. The replacement of cyclophosphamide by azathioprine after the successful induction of remission has been demonstrated not to increase the rate of relapse compared with continued cyclophosphamide. In patients with early antineutrophil cytoplasm antibody associated vasculitides without critical organ manifestations low-dose methotrexate can replace cyclophosphamide for induction treatment with similar remission rates. As the early discontinuation of immunosuppressive treatment is associated with unacceptably high relapse rates, however, treatment for the maintenance of remission is mandatory. Besides azathioprine, leflunomide and methotrexate were efficacious in preventing relapses in Wegener's granulomatosis. Data on anti-cytokine therapy in Wegener's granulomatosis are controversial, possibly related to differences in study design. Open-label clinical studies suggest a beneficial effect of infliximab in addition to standard therapy in refractory Wegener's granulomatosis. In contrast, a recent randomized controlled trial showed that etanercept in addition to standard therapy, with the subsequent tapering of standard medications, is not effective for the maintenance of remission. SUMMARY: Despite recent progress, the prevention of relapses and treatment of refractory cases remain the greatest challenges in the treatment of Wegener's granulomatosis. PMID- 16344617 TI - Kawasaki disease. AB - PURPOSE OF REVIEW: Kawasaki disease is still a dilemma for pediatricians, especially its atypical or incomplete presentation. The current diagnostic criteria appear inadequate, and the early diagnosis frequently remains challenging with high risk of coronary damage. This review focuses on recent data concerning Kawasaki disease etiology and pathogenesis, and emphasizes the proposal of including altered laboratory values in revised diagnostic criteria aimed at identifying patients with incomplete disease. RECENT FINDINGS: Despite a timely treatment, approximately 15% of children have persistent or recurrent fever, thus requiring additional intravenous immunoglobulin, corticosteroids, immunosuppressant, and eventually biologic drugs. Recent cardiologic tests strongly suggest that endothelial damage may lead to coronary stenosis and myocardial ischemia over time, even in children apparently normal at initial cardiac evaluation. New noninvasive tools are suggested to monitor coronary artery aneurysm outcome. Giant coronary artery aneurysms still represent a severe life-threatening complication. There is as yet no agreement regarding the prevention of thrombosis and stenosis in children with giant aneurysms. SUMMARY: Due to earlier recognition, aggressive medical treatment, and surgical procedures, the quality of life in patients with Kawasaki disease is significantly improved. The identification of children at high risk for coronary artery aneurysms is crucial in order to reduce the occurrence of sudden death in adolescence and early adulthood. High parameters of inflammation, anemia, and low sodium and albumin levels, along with persistent unexplained fever, should alert clinicians to suspect the disease even though all clinical symptoms are lacking. PMID- 16344618 TI - Vasculitis and systemic infections. AB - PURPOSE OF REVIEW: In recent years, many investigators have focused on potential associations between infections and vascular inflammation. We review the principal pathogenic mechanisms that have been implicated for possible roles in the vascular inflammation initiated by infectious agents. We also summarize the most important literature related to this topic. RECENT FINDINGS: A novel theory known as autoantigen complementarity suggests that an infectious agent could trigger antineutrophil cytoplasmic antibody-associated vasculitis. Several recent studies investigating the presence of parvovirus B19 and herpesviruses in temporal arteries with giant cell arteritis have yielded contradictory results. A recent study has identified higher frequency of a novel human virus, the 'New Haven coronavirus', in respiratory secretions of children with Kawasaki disease. Many case reports have suggested potential relationships between human pathogens and vasculitis. SUMMARY: There remains considerable interest in the possibilities of primary vasculitic syndromes caused in some fashion by infection. With the exception of a few well sustained associations - for example hepatitis B or C with known vasculitic syndromes - most of the purported links between microbial agents and primary vasculitides remain speculative. PMID- 16344619 TI - Abdominal aortic aneurysms: an underestimated type of immune-mediated large vessel arteritis? AB - PURPOSE OF REVIEW: To review recent studies on the immune-mediated pathogenesis of abdominal aortic aneurysms, opening a wide field for possible new therapeutic approaches. RECENT FINDINGS: Immune-mediated processes including involvement of neutrophils, interferon-gamma producing T cells and proinflammatory cytokines play an important role especially in the initiation of abdominal aortic aneurysm disease. C-reactive protein was associated with aneurysm size and is possibly produced by the aneurysmal tissue itself. From the clinical perspective, both inflammatory and noninflammatory abdominal aortic aneurysms are associated with various autoimmune diseases. Preliminary data of F-FDG positron emission tomography imaging of abdominal aortic aneurysms suggest focal uptake of F-FDG within the aneurysm wall in patients with either large, rapidly expanding or symptomatic aneurysms that are prone to rupture. Thus basic research findings and clinical research focusing on the underlying immune-mediated mechanisms of abdominal aortic aneurysms will likely pave the way for new medical therapies in the future. In animal models the effects of rapamycin as an immunosuppressive agent, modulation of estrogen receptors by tamoxifen as well as gene therapy using decoy oligonucleotides binding to the transcription factor ets has already proved helpful in decreasing aneurysm expansion rates. SUMMARY: Pathophysiological, immunogenetical and interventional studies support the concept of abdominal aortic aneurysm as an immune-mediated process, which will help to identify more laboratory and imaging signs of development in the future. Further research will now assess the possible benefit of antiinflammatory therapeutic approaches, especially in patients with small abdominal aortic aneurysms. PMID- 16344620 TI - Cryoglobulinemic vasculitis. AB - PURPOSE OF REVIEW: Cryoglobulinemic vasculitis is an immune-complex-mediated systemic vasculitis involving small-medium-sized vessels. A causative role of hepatitis C virus in over 80% patients has been definitively established, with heterogeneous geographical distribution. This review focuses on recent etiopathogenetic, clinico-diagnostic, and therapeutical studies. RECENT FINDINGS: Hepatitis C virus cannot be integrated into the host genome; it may exert a chronic stimulus to the immune system. The interaction between hepatitis C virus envelope protein E2 with B-cell CD 81 receptor may increase the frequency of VDJ rearrangement in antigen-reactive B lymphocytes. One consequence is the activation of various protooncogenes, including anti-apoptotic Bcl-2. The extended B-cell survival is responsible for autoantibody and immune-complex production, including mixed cryoglobulins; some malignancies, mainly B-cell lymphomas, may complicate cryoglobulinemic vasculitis. Environmental or viral/host genetic cofactors should be relevant in the pathogenesis of hepatitis C virus-related diseases. Cryoglobulinemic vasculitis may overlap with other diseases (systemic vasculitides, Sjogren's syndrome, autoimmune hepatitis, lymphoma), which should be carefully considered for a correct diagnosis and treatment. Cumulative survival of cryoglobulinemic vasculitis is significantly lower compared with the general population. Therapeutic strategies for cryoglobulinemic vasculitis include etiologic (antiviral), pathogenetic (cyclophosfamide, rituximab), or symptomatic (steroids, plasmapheresis) treatments, which should be tailored to the individual patient according to the severity/activity of clinical symptoms. SUMMARY: Cryoglobulinemic vasculitis represents a crossroads among autoimmune and lymphoproliferative disorders; as hepatitis C virus infection is the major causative factor, cryoglobulinemic vasculitis is an important model for etiopathogenetic studies of virus-related diseases. PMID- 16344621 TI - Cartilage tissue engineering: its potential and uses. AB - PURPOSE OF REVIEW: The prevalent nature of osteoarthritis, a cartilage degenerative disease that results in the erosion of joint surfaces and loss of mobility, underscores the importance of developing functional articular cartilage replacement. Recent research efforts have focused on tissue engineering as a promising approach for cartilage regeneration and repair. Tissue engineering is a multidisciplinary research area that incorporates both biological and engineering principles for the purpose of generating new, living tissues to replace the diseased/damaged tissue and restore tissue/organ function. This review surveys and highlights the current concepts and recent progress in cartilage tissue engineering, and discusses the challenges and potential of this rapidly advancing field of biomedical research. RECENT FINDINGS: Cartilage tissue engineering is critically dependent on selection of appropriate cells (differentiated or progenitor cells); fabrication and utilization of biocompatible and mechanically suitable scaffolds for cell delivery; stimulation with chondrogenically bioactive molecules introduced in the form of recombinant proteins or via gene transfer; and application of dynamic, mechanical loading regimens for conditioning of the engineered tissue constructs, including the design of specialized biomechanically active bioreactors. SUMMARY: Cell selection, scaffold design and biological stimulation remain the challenges of function tissue engineering. Successful regeneration or replacement of damaged or diseased cartilage will depend on future advances in our understanding of the biology of cartilage and stem cells and technological development in engineering. PMID- 16344622 TI - Modern biologics used in orthopaedic surgery. AB - PURPOSE OF REVIEW: In this review we summarize some of the most recent research in the area of local bone regeneration. These innovations may be relevant in the orthopaedic treatment of patients with rheumatoid arthritis, or other inflammatory arthridities, as such patients often present with inadequate bone stock. RECENT FINDINGS: Bone grafting remains the standard treatment for bone deficiency. Several new approaches, such as the use of concentrated blood products or osteoprogenitor cells in conjunction with grafts, have been developed but remain to be tested clinically. Experimental studies have elucidated important aspects of the biology of bone graft remodeling and osteoprogenitor cell differentiation. Materials that can serve as graft alternatives continue to be developed. Positive experimental findings have resulted from combinations of such materials with osteoprogenitor cells or osteoinductive factors such as bone morphogenetic proteins. SUMMARY: While few studies to date have examined the specific use of these new strategies in the setting of rheumatoid arthritis, many hold promise for patients with rheumatoid arthritis and other inflammatory and metabolic conditions that affect bone quality and quantity. PMID- 16344624 TI - Musculoskeletal and autoimmune manifestations of HIV, syphilis and tuberculosis. AB - PURPOSE OF REVIEW: The HIV pandemic continues to increase at an alarming rate, and is the leading cause of death worldwide from a single pathogen. The number of HIV-1-infected individuals currently exceeds 40 million, the majority of whom live in the developing countries of Asia, sub-Saharan Africa and south America. In the past 5 years, there has concurrently been an increase in the reported cases of tuberculosis and primary and secondary syphilis. This review addresses the musculoskeletal and autoimmune manifestations associated with HIV, syphilis and tuberculosis infections or their treatments. RECENT FINDINGS: During HIV infection the immune system becomes dysfunctional because of the coexistence of immunodeficiency and immune hyperactivity, and a disregulated production or activity of cytokines. Some of these mechanisms explain the development of rheumatic manifestations associated with HIV infection. Highly active antiretroviral therapy changes the course of HIV infection and the spectrum of the HIV-associated rheumatic manifestations. New syndromes such as the immune reconstitution inflammatory syndrome have emerged. HIV, tuberculosis and syphilis infections offer special epidemiological, clinical, and therapeutic challenges. SUMMARY: These observations highlight the complexity and multiplicity of the interactions between the pathogen and host that could result in the development of rheumatic manifestations. PMID- 16344623 TI - Periprosthetic osteolysis: an immunologist's update. AB - PURPOSE OF REVIEW: Inflammation-induced osteolysis is a problem in both inflammatory arthritis and total joint arthroplasty. New drug therapies have been shown to slow, halt, or even reverse the osteolysis associated with inflammatory arthritis. Unfortunately, similar advances in the medical treatment of periprosthetic osteolysis have not occurred. This review will update the state of periprosthetic osteolysis. RECENT FINDINGS: Preliminary results with phase I and II clinical trials with AMG-162, a human IgG2 that binds receptor activator of nuclear factor kappaB (RANK) ligand, have been reported. Based on these results AMG-162 appears to be safe and to have a potent effect on osteoclast function. Based on animal studies, it is expected that regents such as AMG-162 that block RANK-ligand/RANK interaction will have activity in inflammation-induced osteolysis. Volumetric three-dimensional and magnetic resonance imaging scans for detecting and quantifying periprosthetic osteolysis have been validated in cadaver studies. Lymphocytic infiltrates and positive skin tests to cobalt have been found in patients with periprosthetic osteolysis after second generation metal-on-metal prostheses. These findings again raise the question of whether metal allergy may contribute to implant failure in these patients. A new subset of T helper cells that are neither Th1 nor Th2, but secrete a unique pattern of cytokines including IL-17, has recently been discovered. The importance of these cells in modifying particle-induced osteolysis remains to be determined. SUMMARY: There have been significant advances in our understanding of periprosthetic osteolysis, imaging technology to quantify osteolysis, and drug development. The time now seems ripe to translate these advances in clinical trials. PMID- 16344625 TI - Musculoskeletal manifestations and autoimmune diseases related to new biologic agents. AB - PURPOSE OF REVIEW: The anti-tumor necrosis factor agents are now widely used in the management of patients with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis and juvenile inflammatory arthritis. One of the most common observations made during their use is the development of autoantibodies. The purpose of this paper is to review this phenomenon and its clinical implications. RECENT FINDINGS: While the development of different autoantibodies is a common encounter, rare cases of lupus-like syndromes have been reported. On the other hand, a variety of immune-mediated clinical manifestations have been described, including vasculitis and demyelinating syndromes. Rare cases of cytopenia and non specific lung injuries have also been reported. SUMMARY: While these clinical complications are rare and isolated events, clinicians must be aware of their occurrence. The experience with the anti-tumor necrosis factor agents is rather short and new, unusual immune-mediated complications may still appear. Clinicians should be prepared to recognize them. PMID- 16344626 TI - Celiac sprue (the great modern-day imposter). AB - PURPOSE OF REVIEW: To review the current epidemiological information on celiac disease and the various presentations and associated. RECENT FINDINGS: Epidemiologic studies reveal celiac disease to be common, occurring in approx. 1% of the population. It is being diagnosed worldwide, even in developing countries. The classic mode of presentation has become less common, with diarrhea or a malabsorption syndrome as the mode of presentation in fewer than 50% of individuals. The other major modes of presentation are iron-deficiency anemia, osteoporosis, screening of family members, or incidentally at endoscopy done for dyspepsia or reflux. Neurological presentations may include peripheral neuropathy or ataxia. Arthritis is commonly found in patients with celiac disease when systematically sought. Patients often have a previous diagnosis of irritable bowel syndrome. Autoimmune diseases occur more frequently (three to ten times more) in those with celiac disease than the general population. However, this increased incidence of autoimmune diseases is not prevented by early diagnosis of celiac disease. SUMMARY: We will review the various associated diseases/presentations of celiac disease. The heterogeneity of the symptoms can make the diagnosis challenging and certainly the great modern-day imposter. PMID- 16344628 TI - Bibliography. Current world literature. Vasculitis syndromes. PMID- 16344627 TI - Familial Mediterranean fever and the other autoinflammatory syndromes: evaluation of the patient with recurrent fever. AB - PURPOSE OF REVIEW: The aim of this article is to summarize recent clinical, genetic and pathophysiologic findings of familial Mediterranean fever and several of the other systemic autoinflammatory diseases, a recently recognized group of disorders characterized by seemingly unprovoked inflammation but lacking high titer autoantibodies. Genetic and clinical tools are improving the ability of the clinician to better approach patients with periodic fever and inflammation. RECENT FINDINGS: The spectrum of reported genetic mutations and susceptible ethnicities for the hereditary periodic fever subset of the autoinflammatory diseases has continued to expand. At the same time, the pathogeneses of many of these diseases are now understood to involve different aspects of a common pathway, largely affecting inflammatory cascades related to IL-1 or tumor necrosis factor-alpha. Three of these diseases which have been grouped as the cryopyrin-associated periodic syndromes result from defects in the same gene, and all three appear to respond well to anti-IL-1 therapy although controlled trials are still in progress. In addition, cytokine-based therapies are also now under investigation for hyperimmunoglobulinemia D with periodic fever syndrome and pyogenic sterile arthritis, pyoderma gangrenosum, and acne syndrome. SUMMARY: The identification of the genes and proteins mutated in many of the autoinflammatory diseases has broadened our understanding of the regulation of inflammation and the immune system, and provided the basis for the use of targeted therapies in these syndromes. We propose an algorithm for the evaluation of a patient with periodic fever, taking into account the patient's age, ethnicity, symptoms and signs, and results from laboratory and genetic testing. PMID- 16344631 TI - Patient-provider communication in chronic illness: a health promotion window of opportunity. AB - As chronic illness takes its rightful place as an imperative for planners and policy makers globally, the obligation to provide appropriate self-care management support and promote optimal health among the chronically ill is attracting considerable attention. A generation of insider research into the chronic illness experience has yielded powerful evidence of the critical importance of communication between the chronically ill and those who provide their healthcare services. In this article, insights gained from the body of chronic illness research shape recommendations for attitudinal and structural adjustments that might steer our course toward promoting improved health for those affected by a chronic disease. PMID- 16344632 TI - Health promotion for persons with disabilities: what does the literature reveal? AB - A search of MEDLINE and CINAHL databases for research on disability and health promotion was done. Twenty-three articles were categorized into 1 of 3 areas: the meaning of health and health promotion, factors that contribute to health and health promotion, and health promotion interventions. Overall, health and health promotion were inductively defined concepts that emphasized function, relationships, and a positive mental attitude. Barriers to health promotion were frequently reported, fatigue being most common. Moreover, better health outcomes were reported when people with disabilities engaged in health-promoting behaviors. There were few interventions found, with only 1 being a randomized clinical trial. PMID- 16344633 TI - Is health promotion relevant across cultures and the socioeconomic spectrum? AB - Is health promotion a White middle-class phenomenon that people from other cultures and classes do not regard as important? When implementing health promotion initiatives, are healthcare providers making assumptions that are not valid for other cultural or socioeconomic groups? How do people of various cultures and classes perceive health and health promotion? To explore these questions, this article reviews some of the relevant literature on culture and class in relation to health promotion, exploring issues foundational to the effectiveness of health-promotion programs and pertinent to delivering health promotion interventions to ethnic, racial, and cultural minorities and poor populations. Health promoters are encouraged to consider the social determinants of their patients' health and tailor programs on the basis of their patients' motivations and resources. PMID- 16344634 TI - Building health promotion interventions for persons with chronic disabling conditions. AB - There is a growing number of persons with chronic disabling conditions and a concurrent interest and need for health-promotion interventions to prevent disability and promote quality of life within the context of chronic conditions. Most often researchers/clinicians build their own intervention focusing on selected dimensions of health promotion for a specific population. This article recommends an alternative process of building health-promotion interventions for specific groups through the adaptation of content and processes of well-developed intervention frameworks with sound theoretical and empirical support. This efficient approach enhances the likelihood that new interventions will prove to be effective and enhance the quality of life for persons with chronic disabling conditions. PMID- 16344635 TI - Preventive health screening for breast and cervical cancer and osteoporosis in women with physical disabilities. AB - Although the Americans With Disabilities Act was enacted 15 years ago in an effort to improve access of people with disabilities to a broad range of services, women with physical disabilities continue to receive less preventive health screening than women without disabilities and less than is recommended. Furthermore, women with more severe disabilities undergo less screening than those with mild or moderate severity of disability. This article reviews findings of studies on health screening for breast and cervical cancer and osteoporosis in women with physical disabilities and identifies practice and research implications on the basis of those findings to improve the health status of women with physical disabilities. PMID- 16344636 TI - Achieving effective health promotion for women with disabilities. AB - The field of health promotion has yet to acknowledge the unique needs of women with disabilities, a population representing approximately 1 of 5 women in the United States. Compared with women without disabilities, women with disabilities have critical needs for evidence-based health promotion services. Women with disabilities face a lack of access to multitudinous opportunities for maintaining and improving their overall health. Inaccessible exercise equipment and other disability-related barriers discourage women with physical disabilities from engaging in health-promoting behaviors. This article identifies 10 essential elements for achieving effective health promotion research and interventions for women in this population. PMID- 16344637 TI - Supporting health promotion in adults with cancer. AB - Health promotion includes motivating people to embrace behaviors that both improve their emotional and physical quality of life and reduce their risk for premature morbidity and mortality. Although the majority of adults treated for cancer in the United States live free of disease for many years, many survivors experience lasting side effects and complications of treatment. Others encounter recurrences or new cancers that require additional treatment. Cancer is an incentive for many survivors to make health-promoting lifestyle changes. This article delineates the need for health promotion among adult cancer survivors, describes a cancer support model that facilitates personal behaviors that promote health, and discusses design implications for cancer supportive services. PMID- 16344638 TI - Sexual health promotion in adolescents with chronic health conditions. AB - Sexual health promotion is essential to the health and well-being of adolescents, including those with chronic health conditions. The purposes of this article are to define sexual health and to describe the physical, cognitive, psychosocial, and spiritual domains of adolescent development, with special focus on the sexual health needs of adolescents with various chronic health conditions. The article concludes with several suggestions for healthcare professionals, parents, and teachers in addressing sexual health promotion in this vulnerable population. PMID- 16344639 TI - Measuring health among people with disabilities. AB - The evolution in our understanding of health has profound implications for people with disabilities. The purpose of this article is to explore how health has been defined and measured for people with disabilities. This article concludes with recommendations to improve measurement of health among people with disabilities. These recommendations include clarifying conceptual and operational definitions of health with input from people with disabilities, exploring whether existing health measures might be modified to encompass environmental supports for staying healthy and avoiding secondary conditions, and conducting population-based studies of health status of people with disabilities. PMID- 16344640 TI - Physical activity measurement in persons with chronic and disabling conditions: methods, strategies, and issues. AB - Measuring the physical activity of persons with chronic and disabling conditions presents complexities related to measuring instruments, the intensity of the activity being measured, the population being measured, and individual behavior and health status. They often have limitations in mobility that do not preclude physical activity but contribute to the complexity of measuring it, such as slow or altered gait, inability to walk, and the need for assistive devices. This article reviews currently available ways to measure physical activity, describes strengths and weaknesses of various measures, and provides examples of complexities in measuring physical activity in people who move differently. PMID- 16344641 TI - Is the language of research the real barrier? PMID- 16344643 TI - Barriers to research use. PMID- 16344644 TI - Enhancing collaboration with academic partners. PMID- 16344645 TI - Relationship and results-oriented healthcare: evaluate the basics. PMID- 16344646 TI - Creating a favorable practice environment for nurses. AB - Chronic nurse turnover and attendant personnel shortages threaten patient care. Inadequate pay and benefits are primary factors leading to nurses' job dissatisfaction and, subsequently, to turnover. However, recent meta-analyses suggest that a favorable practice setting can improve nurses' satisfaction and minimize turnover. The practice environment's potential to positively influence nurses' job satisfaction is analyzed in a large academic medical center. Nurse supervisors, job characteristics, management style, and service quality emphasis- not just money and benefits--surface as key factors associated with a positive practice milieu. Implications for raising nurses' job satisfaction, reducing turnover, and improving care delivery are discussed. PMID- 16344647 TI - The RWJ Executive Nurse Fellows Program, Part 2: Mentoring for leadership success. AB - This article is the second in a 3-part series describing the RWJ Executive Nurse Fellows Program, an advanced leadership program for nurses in senior executive roles who aspire to help lead and shape the US healthcare system of the future. Part 1 (October 2005) described the program, its core leadership competencies, and the primary components. This article discusses the mentor experience that is a cornerstone of the 3-year fellowship program. Fellows are encouraged to have this experience with senior-level executives outside of healthcare in order to broaden their leadership perspectives. Examples of these mentor experiences are described from the viewpoints of both fellows and mentors, including successes, challenges, and lessons learned. Part 3 (February 2006) will explain how fellows are required to create a business plan for their leadership project because it is so important for nurse leaders to offer a strong business case for proceeding with anew initiative, service, or program. PMID- 16344648 TI - Nursing diagnoses, diagnosis-related group, and hospital outcomes. AB - BACKGROUND AND OBJECTIVE: There are no nursing centric data in the hospital discharge abstract. This study investigates whether adding nursing data in the form of nursing diagnoses to medical diagnostic data in the discharge abstract can improve overall explanation of variance in commonly studied hospital outcomes. METHOD: A retrospective analyses of 123,241 sequential patient admissions to a university hospital in a Midwestern city was performed. Two data sets were combined: (1) a daily collection of patient assessments by nurses using nursing diagnosis terminology (NDX); and (2) the summary discharge information from the hospital discharge abstract including diagnosis-related group (DRG) and all payer refined DRG (APR-DRG). Each of 61 daily NDX observations were collapsed as frequency of occurrence for the hospital stay and inserted into the discharge abstract. NDX was then compared to both DRG and APR-DRG across 5 hospital outcome variables using multivariate regression or logistic regression. RESULTS AND CONCLUSIONS: In all statistical models, DRG, APR-DRG, and NDX were significantly associated with the 5 hospital outcome variables (P <.0001). When NDX was added to models containing either the DRG or the APR-DRG, explanatory power (R2) and model discrimination (c statistic) improved by 30% to 146% across the outcome variables of hospital length of stay, ICU length of stay, total charges, probably of death, and discharge to a nursing home (P <.0001). The findings support the contention that nursing care is an independent predictor of patient hospital outcomes. These nursing data are not redundant with the medical diagnosis, in particular, the DRG. The findings support the argument for including nursing care data in the hospital discharge abstract. Further study is needed to clarify which nursing data are the best fit for the current hospital discharge abstract data collection scheme. PMID- 16344649 TI - Improving nursing unit teamwork. AB - A lack of teamwork among nursing staff affects care delivery and unit operations. Barriers present in the structure of a typical patient care unit that make it extremely difficult to achieve a high level of teamwork include large team size, lack of familiarity, instability of the work force and assignments, the absence of a common purpose and destiny, and an inhibiting physical environment. The authors discuss strategies to overcome these obstacles to teamwork. PMID- 16344650 TI - Logic models as aids in managing health programs. AB - Logic models, which are graphic depictions of how programs are intended to operate, can be very useful devices for improving the management of health programs in two important ways. They can assist program managers to perform the core management activities of strategizing, designing, and leading other program participants in an integrated manner. Logic models also can assist managers in establishing and maintaining good program stakeholder relationships. PMID- 16344651 TI - Retaining older hospital nurses and delaying their retirement. AB - Delaying the retirement of aging hospital nurses and making work-environment changes to support an older bedside nursing population are critical to stemming the exodus of experienced nurses from the work force. These issues were examined in this descriptive survey study of 1,553 hospital-based nurses in central New England. PMID- 16344654 TI - Premonitory Urge for Tics Scale (PUTS): initial psychometric results and examination of the premonitory urge phenomenon in youths with Tic disorders. AB - Although motor tics and/or vocal tics are the defining features of chronic tic disorder (CTD) and Tourette syndrome (TS), older youths and adults often report their tics to be preceded by an unpleasant sensation or "premonitory urge." While premonitory urge phenomena may play an important role in behavioral interventions for CTD/TS, standardized assessments for premonitory urges do not exist. The current study of 42 youths with TS or CTD presents initial psychometric data for a new, brief self-report scale designed to measure tic-related premonitory urges. Results showed that the Premonitory Urge for Tics Scale (PUTS) was internally consistent (alpha = .81) and temporally stable at 1 (r = 0.79, p < .01) and 2 (r = 0.86, p < .01) weeks. PUTS scores were also correlated with overall tic severity as measured by the Yale Global Tic Severity Scale (YGTSS; r = 0.31, p < .05) and the YGTSS number (r = 0.35, p < .05), complexity (r = 0.49, p < .01), and interference (r = 0.36, p < .05) subscales. Finally, an examination of the psychiatric correlates of the premonitory urge phenomenon yielded significant correlations between the PUTS and the Child Behavior Checklist (CBCL) anxiety/depression (r = 0.33, p < .05), and withdrawal (r = 0.38, p < .05) subscales as well as the Children's Yale-Brown Obsessive Compulsive Scale (CYBOCS; r = 0.31, p < .05). However, a cross-sectional examination of the data showed that the psychometric properties of the PUTS were not acceptable for youths 10 years of age and younger. Likewise, significant correlations found between the YGTSS subscales, CBCL subscales, CYBOCS, and the PUTS did not emerge in this younger age group. The clinical and theoretical implications of these findings are discussed. PMID- 16344656 TI - Nocturnal enuresis and psychosocial problems in pediatric sickle cell disease and sibling controls. AB - To evaluate current and lifetime prevalence rates of nocturnal enuresis and psychosocial problems among children with sickle cell disease (SCD) in comparison with sibling controls, a structured interview and the Pediatric Symptom Checklist were administered to primary caretakers regarding 126 of their children aged 5 to 17 with SCD and 47 sibling controls. Lifetime rates of enuresis among children with SCD were comparable to similar studies, and exceeded population prevalence and sibling control rates. In addition, enuretic children had higher levels of total psychosocial problems on the Pediatric Symptom Checklist regardless of group status, although patterns of subscale differences varied by group and enuresis history after controlling for child age. These findings replicate and extend previous findings and provide further evidence to support a need for monitoring of hydration levels and screening for psychosocial problems among children with SCD and enuresis, as well as evaluation of the psychometric properties of psychosocial screening measures and identification of efficacious treatments for enuresis in children with SCD. PMID- 16344657 TI - Validity of the Clinical Adaptive Test (CAT)/Clinical Linguistic and Auditory Milestone Scale (CLAMS) as a screening instrument for very low birth weight infants in Taiwan. AB - The authors investigated the validity of the Clinical Adaptive Test (CAT)/Clinical Linguistic and Auditory Milestone Scale (CLAMS) for developmental screening and early prediction in very low birth weight (VLBW) infants, using the Bayley Scales of Infant Development II (BSID-II) as a reference standard. A total of 808 VLBW infants discharged from neonatal intensive care units in Taiwan from January 1995 to December 1997 were enrolled and followed up at the corrected ages of 6, 12, 18, and 24 months. The CAT/CLAMS and BSID-II were performed separately during each visit. The CAT/CLAMS showed strong concurrent correlations (r = 0.51 0.86, p < .0001) with BSID-II mental developmental index (MDI), and had high conegativity scores (96.7%-100%) but low copositivity scores (30.0%-45.3%) for detecting developmental delay. At 24 months of age, the CAT/CLAMS had better predictive validity for significant (MDI <70) rather than borderline (MDI = 70 84) developmental delay. The authors conclude that the CAT/CLAMS is not an appropriate developmental instrument for screening and early prediction in VLBW infants in Taiwan. PMID- 16344658 TI - Rebuilding a region: 200 billion dollars restoring communities: priceless. PMID- 16344659 TI - In pursuit of excellence: the third phase of the Journal of Developmental and Behavioral Pediatrics, 1997-2002. PMID- 16344660 TI - Scott: an 11-year-old boy with repetitive lying. PMID- 16344661 TI - Neurodevelopmental outcomes of infants born prematurely. AB - Long-term follow-up of infants born prematurely is necessary to determine neurodevelopmental outcomes, particularly with the expansion of interest from major disabilities to high prevalence/low severity dysfunctions. Models of pathogenesis include changes due to developmental disruptions and to injury, the magnitude and type of change influenced by the infant's age, and central nervous system recovery and reorganization. Alterations in neurogenesis, migration, myelination, cell death, and synaptogenesis occur even in the absence of insult. Despite increased knowledge regarding these processes, the functional significance of brain abnormalities is unclear. Because of methodologic problems in follow-up studies, it is difficult to characterize outcome definitively. Nonetheless, an acceptable degree of agreement across studies is found with regard to specific neurodevelopmental outcomes: motor/neurologic function, visuomotor integrative skills, IQ, academic achievement, language, executive function, and attention-deficit hyperactivity disorder/behavioral issues. In general, children born prematurely have more problems in these areas than do their normal birth weight counterparts. Suggestions for improved analyses and clarification of outcomes include use of cluster analysis, structural equation modeling, growth curve analysis, developmental epidemiologic approaches, and better control of background variables using risk indexes and factor scores. Better assessment techniques measuring functions documented to be at higher risk of problems are discussed. PMID- 16344662 TI - Parental beliefs and decision making about child and adolescent immunization: from polio to sexually transmitted infections. AB - Over the past 150 years, innovations in immunization practices have dramatically improved the health of children, and parents are increasingly asked to consider and accept new childhood vaccines. We present a conceptual model to frame a review of research on the role of parental attitudes and beliefs in decision making about child and adolescent immunization and describe the historical context of vaccine-related decision-making research. This review focuses on theory-based Social-environmental and parent-specific personal factors as potential influences on vaccine decision making. Relevant Social-environmental issues discussed include media coverage of vaccines, perceived social norms, and the persuasive influence of peer groups. Health care provider recommendations are presented as an exemplar of factors related to the family's interface with the health care system. Personal factors addressed include parental health beliefs, attitudes, and knowledge related to vaccine preventable diseases and immunization, as well as cognitive heuristics that are employed in the decision making process (e.g., omission bias, protected values, framing of information). Last, promising directions for research and suggestions for clinical practice are presented. PMID- 16344666 TI - Pediatric clinical research in benign hematology: a dilemma? PMID- 16344668 TI - Parents' assessment of risk in sickle cell disease treatment with hydroxyurea. AB - Hydroxyurea is the only medication shown to reduce the severity of sickle cell disease (SCD), but its long-term risks are unknown. Families of 58 children with SCD were interviewed on their perception of the potential risks of hydroxyurea treatment, and whether disease severity was a major factor in their treatment decision. Parents rated the severity of their child's SCD (86% as severe, 26% moderate, and 47% mild) and then the highest levels of risk that they would tolerate for both potential cancer (range 1/1,000 to 1/2) and potential birth defects (range 1/1,000 to 1/3) to benefit their child with hypothetical hydroxyurea treatment. Parents of 29 of the 58 children were unwilling to take any cancer risk whatsoever as a potential side effect of treatment, and 29 of 58 (not all the same parents) were unwilling to take any risk for birth defects, including half the children with severe SCD. For those families who did accept some risk, higher acceptable risk correlated with higher disease severity in the child (P = 0.04). The study's mixed findings highlight the complexities of risk perception and suggest that future studies of risks and benefits from the parental viewpoint are needed as candidate therapies are developed for SCD. PMID- 16344667 TI - Attitudes of children with leukemia toward repeated deep sedations with propofol. AB - Procedural sedation is generally recommended for children requiring repeated painful diagnostic or therapeutic procedures. A child with leukemia undergoes an average of 20 procedures such as lumbar puncture and bone marrow aspiration through the course of illness. No data are currently available about the psychological impact of repeated sedations on children. The objective of this study was to evaluate the attitudes of patients with leukemia toward repeated deep sedations using propofol. A questionnaire addressing sedation-related distress was given to 30 children with leukemia. Procedure-related distress was evaluated using the Amended Observational Scale of Behavioural Distress. Another questionnaire concerning the same issues was given to an historical group of 39 children who had undergone painful procedures without sedation in previous years. Fear and distress were significantly reduced in the sedation group compared with the historical one. Fear of sedation was reported by 17% of children of this group. Distressed behavior was observed in 27%. In conclusion, sedation-related distress was observed in a subgroup of patients; in these cases, specific strategies could be considered to reduce sedation-related distress. PMID- 16344669 TI - Auditory and visual toxicity during deferoxamine therapy in transfusion-dependent patients. AB - Deferoxamine is a chelating agent that has extended the life expectancy of patients with thalassemia. In the 1980s, many investigators reported otologic and visual toxicity caused by deferoxamine. In July 1999 and 2 years later, the authors performed audiologic and ophthalmologic assessments in 30 transfusion dependent patients receiving deferoxamine therapy (40-50 mg/kg per dose, subcutaneously for 8-10 hours, 4-7 days per week). In 1999, six patients (20%) had deferoxamine-related hearing impairment (>25 dB), all at high frequencies. Because the authors believed the benefits of chelation therapy outweighed the risk of ototoxicity, the dose of deferoxamine was not reduced. Two years later, the hearing impairment had not progressed in any of the patients. There was no association between ototoxicity and ferritin level. No patients had abnormalities of visual acuity or funduscopy in either 1999 or 2001. Based on this experience, deferoxamine at doses lower than 50 mg/kg/d was safe for the eyes and slightly toxic to the ears. The ototoxicity probably relates to individual susceptibility. Regular monitoring of auditory function and close follow-up of abnormal findings are recommended. According to this limited experience, reducing the dose or withdrawing deferoxamine might not be necessary if the hearing loss is stable in the face of ferritin levels above 2,000 ng/mL. Because of the relatively small patient numbers, more data are needed to confirm these conclusions. PMID- 16344670 TI - Increased pulmonary artery pressures among adolescents with sickle cell disease. AB - The prevalence of pulmonary hypertension (PHT) among adolescents with sickle cell disease (SCD) is unknown. A tricuspid regurgitant (TR) jet peak velocity of 2.5 m/s or more is a screening test for PHT. The authors retrospectively reviewed echocardiograms and clinical data of adolescents followed at the Texas Children's Sickle Cell Center. Of 80 evaluable adolescents with SCD and echocardiogram data, 21 (26%) had a TR jet velocity of 2.5 m/s or more. Of these 21 patients with PHT, 12 (57%) had an echocardiogram performed during an inpatient stay for vaso occlusive crisis (n = 6), acute chest syndrome (n = 4), fever (n = 1), or seizures (n = 1), and 9 (43%) had an echocardiogram performed as an outpatient in a baseline state of health. Elevation of pulmonary artery pressures was common in this adolescent cohort, but clinical symptoms were rare. Prospective study is warranted to determine the prevalence and course of elevated pulmonary artery pressures in this age group. PMID- 16344671 TI - West Nile virus infection in a teenage boy with acute lymphocytic leukemia in remission. AB - West Nile Virus (WNV) infection is an important cause of encephalitis. Although the medical literature contains examples of WNV encephalitis in susceptible, mainly elderly, immunocompromised hosts, few case reports have described pediatric cases. The authors describe an adolescent with acute lymphocytic leukemia and WNV encephalitis. Surveillance studies indicate an increase in WNV activity. Physicians need to be aware of WNV activity in their community and consider WNV as a potential source of infection. PMID- 16344672 TI - Idiopathic hypereosinophilic syndrome in children: report of a 7-year-old boy with FIP1L1-PDGFRA rearrangement. AB - Idiopathic hypereosinophilic syndrome (HES) in children is a very rare disorder; certain clinical differences with adult HES have been described, with no pediatric case with the imatinib-responsive FIP1L1-PDGFRA fusion gene reported to date. The authors describe the clinical course of three children with HES in whom FIP1L1-PDGFRA fusion gene was studied and report the first child with this rearrangement. PMID- 16344673 TI - Multifocal Castleman disease in pediatrics: case report. AB - Multicentric Castleman disease is a rare lymphoproliferative disorder of unknown cause. It is especially rare in children. The authors describe a 4-year-old girl that presented with protracted fever, hypoalbuminemia, thrombocytopenia, anasarca, and disseminated lymphadenopathy. Despite a comprehensive evaluation, her diagnosis remained elusive for several weeks. Eventually, a lymph node biopsy showed the presence of multicentric Castleman disease of mixed type. Due to her relatively poor prognosis and severity of the disease, she was treated with combination chemotherapy consisting of cyclophosphamide, vincristine, doxorubicin, and prednisone. She tolerated her therapy well and is currently free of disease at 36 months of follow-up. PMID- 16344674 TI - Bilateral adrenal cystic neuroblastoma with hepatic and splenic involvement in a newborn. AB - Bilateral cystic adrenal neuroblastoma is an unusual variant of neuroblastoma, and only two cases have been reported in the neonatal period until now. The authors describe a newborn with splenic and hepatic involvement of bilateral adrenal cystic neuroblastoma. PMID- 16344675 TI - Simultaneous control of third-degree graft-versus-host disease and prevention of recurrence of juvenile myelomonocytic leukemia (JMML) with 6-mercaptopurine following fulminant JMML relapse early after KIR-mismatched bone marrow transplantation. AB - The authors describe a young boy with juvenile myelomonocytic leukemia (JMML) who relapsed 45 days after HLA and killer immunoglobulin-like receptor (KIR) mismatched unrelated donor bone marrow transplant (MMUD-BMT) and subsequently developed life-threatening graft-versus-host disease (GvHD). Treatment with 6 mercaptopurine (6-MP) appeared to control severe GvHD and possibly prevented recurrence of leukemic relapse. PMID- 16344676 TI - p16 inactivation associated with aggressive clinical course and fatal outcome in TEL/AML1-positive acute lymphoblastic leukemia. AB - The authors describe a 7-year-old boy with TEL/AML1-positive pre-B acute lymphoblastic leukemia, with hemizygous 9p21 deletion at presentation and no p16(INK4A) protein expression. Despite an initial response to a standard chemotherapy regimen, the patient suffered two hematologic relapses and died 34 months after diagnosis. The authors discuss the possibility that complete p16(INK4A) gene inactivation may adversely modify the prognostic significance of TEL/AML1 fusion in childhood acute lymphoblastic leukemia, and present evidence from clinical and in vitro observations in favor of this assumption. PMID- 16344677 TI - Abdominal pain in a patient with osteosarcoma. AB - Abdominal pain often occurs in patients receiving chemotherapy. The authors describe a patient with osteosarcoma who developed severe right-sided abdominal discomfort several days after being admitted for fever, neutropenia, and mucositis. Unexpectedly, the patient's pain was not therapy-related, but rather was caused by a midline pelvic mass. PMID- 16344678 TI - Multimodal treatment using surgery, radiotherapy, and chemotherapy in a patient with a perivascular epithelioid cell tumor of the uterus. AB - Perivascular epithelioid tumor (PEComa), a recently defined tumor, is a very rare disease affecting various organs, most often the uterus. This tumor displays a variety of histologic and clinical features and at this point is regarded as a tumor with uncertain malignant potential. A 9-year-old girl with abdominal pain and vaginal spotting was diagnosed with PEComa of the uterus with metastasis. She received chemotherapy comprising vincristine, ifosfamide, and doxorubicin, as well as radiotherapy after surgery. After this multimodal treatment, there was no evidence of recurrence or further metastasis. She remains disease-free 1.5 years after her initial diagnosis. PEComa of the uterus displaying malignant characteristics may have a more favorable response to more aggressive therapy. PMID- 16344679 TI - Diffuse thoracic lymphangiomatosis with disseminated intravascular coagulation in a child. AB - An 8.5-year-old girl who presented with chronic cough and hemoptysis underwent a CT scan of the chest showing diffuse mediastinal and parenchymal infiltration and pleural effusion, and laboratory findings showed disseminated intravascular coagulation. Disseminated lymphangiomatosis was diagnosed after an open-lung biopsy. She was treated by systemic steroids, interferon, tamoxifen, chemotherapy, and radiation but died of respiratory failure and disseminated intravascular coagulation 2 years after the diagnosis. This patient represents a rare presentation of diffuse pulmonary lymphangiomatosis together with disseminated intravascular coagulation, involving both the mediastinum and pulmonary parenchyma, in a child. PMID- 16344680 TI - Thrombolysis of venous and arterial thrombosis by catheter-directed low-dose infusion of tissue plasminogen activator in children. AB - Thrombolytic therapy is a well-defined treatment option for arterial and venous thrombosis in adults. In contrast, uniform recommendations regarding the indication, route of administration, and dosing of thrombolytic therapy in children are not available. The authors report the successful resolution of bilateral pulmonary embolism and popliteal artery thrombosis in an 11-year-old girl and 13-year-old girl, respectively, by catheter-directed thrombolysis with low-dose recombinant tissue plasminogen activator. Catheter-directed low-dose thrombolysis is an efficient treatment option for severe venous and arterial thrombosis in children. PMID- 16344681 TI - Superior vena cava thrombus treated with low-dose, peripherally administered recombinant tissue plasminogen activator in a child: case report and review of the literature. AB - Thrombotic complications in pediatrics are more common today due to our ability to treat complicated diseases. In pediatric patients where thrombolytic therapy is indicated, the lack of evidence-based medicine forces practitioners to extrapolate dose recommendations for recombinant tissue plasminogen activator (rt PA) from adult studies. This often results in a high rate of major complications. Low-dose rt-PA is an option that is effective and safe. The authors describe a child with septic thrombus of the superior vena cava who was successfully treated with low-dose, peripherally administered rt-PA, and review the efficacy and safety of standard- versus low-dose rt-PA in children. A Medline search was completed for the use of standard- and low-dose rt-PA in children with thromboembolic disease. Five studies large enough to present efficacy and safety data on standard- and/or low-dose t-PA use in children and neonates were analyzed. Two studies using low-dose rt-PA reported efficacy rates equal to or greater than those reported for standard-dose regimens. Rates of major complications were approximately 5% for low-dose regimens and up to 40% for standard-dose ones. Low-dose, peripherally administered rt-PA is safe and may be as effective as standard-dose rt-PA. A randomized controlled trial should be done to confirm this assumption. PMID- 16344682 TI - Pyrite or true gold? PMID- 16344683 TI - Autologous transplantation of fascia into the vocal fold: long-term result of type-1 transplantation and the future. AB - OBJECTIVES: Since 1997, we have performed the autologous transplantation of fascia into the vocal fold (ATFV) procedure on cases of sulcus vocalis. In what follows, we report the long-term results of our new surgical approach and discuss the role of these transplantations. We also review and report some complications that can be caused by ATFV. Finally, we discuss the ATFV technique as a contribution to the phonosurgery of the future. STUDY DESIGN: Prospective study. METHODS: We were able to obtain long-term results from 10 volunteer cases (2 female and 8 male, age: 15-71, mean 46.5 years old) who could be followed up for at least 3 years after transplantation. All were cases of pathologic sulcus vocalis.We measured maximum phonation time (MPT) and carried out pre- and postsurgical clinical observation and laryngeal stroboscopy in all cases. These measurements and observations were made before the ATFV and at 6 months, 1 year, 2 years, and 3 years after surgery. RESULTS: In stroboscopic observation 1 year after the ATFV, satisfactory glottal closure and excellent mucosal wave were observed for all cases, and there was no case with hyperadduction of the false vocal folds. MPT measures remained at an improved level 2 years and 3 years after the transplantations. Paired-sample t tests showed that the improvement relative to preATFV levels was significant for all postsurgical measurements up to 3 years. CONCLUSIONS: We conclude that ATFV is a successful surgical procedure for sulcus vocalis and scarred vocal folds. Other phonosurgical clinical applications may also be envisioned. PMID- 16344684 TI - Forge the connection between technology and compassion. PMID- 16344685 TI - How well do you know your patient population? AB - A firm knowledge of patient population characteristics enables accurate competency assessment. PMID- 16344687 TI - Best-practice protocols: VAP prevention. AB - A standardized approach to care delivery helps intensive care units reduce VAP care complications. PMID- 16344688 TI - RRTs and electronic code documentation: advancing in-hospital resuscitation practices. AB - Review new methods for preventing arrest and improving survival. PMID- 16344689 TI - Integrated technologies transform telemetry. AB - Current monitoring and alarm notification technologies can help merge biomedical, information, and telecommunication systems into completely new configurations. PMID- 16344690 TI - Gain new perspectives in improved guidelines and practices. AB - Target open communication and a renewed focus on prevention for better infection control. PMID- 16344691 TI - Is nursing ready for 2006? AB - From the worsening nursing shortage to changes in Medicare's structure, nursing has much to do in preparation for next year. PMID- 16344692 TI - Harness technology for sustainable regulatory compliance. AB - Translate each new regulation into a process change that can be implemented and sustained economically, with reasonable effort. PMID- 16344693 TI - Pump away high-risk infusion errors. AB - New intravenous pumps help increase safety measures and improve cost effectiveness. PMID- 16344694 TI - Resource management: what's right for you? AB - Explore the benefits of both a decentralized staffing and scheduling model and a centralized model. PMID- 16344697 TI - Is employee discipline the solution for patient safety? AB - Explore alternative solutions to discipline and address ways to achieve patient safety without penalizing employees. PMID- 16344699 TI - More IT, better patient care? PMID- 16344698 TI - What's your nursing philosophy? AB - Nurses who are self-motivated, continue their education, and are active in professional organizations enjoy career satisfaction and provide better healthcare. PMID- 16344702 TI - Healthy People 2010: a New Year's resolution for us all. PMID- 16344704 TI - The Patient Safety and Quality Improvement Act of 2005. PMID- 16344705 TI - Recognizing and managing anemia. PMID- 16344707 TI - Coarctation of the aorta: a critical catch for newborn well-being. PMID- 16344709 TI - Stepping up foot injury diagnosis: Jones, Lisfranc, and Charcot. PMID- 16344711 TI - Guide to care for patients. Anemia. PMID- 16344710 TI - Does RN experience relate to NP clinical skills? AB - Generally, a registered nurse (RN) wishing to become a nurse practitioner (NP) enters a Master's NP program with at least 1 year of experience. However, some schools admit students directly to graduate NP study without prior nursing experience. Are inexperienced nurses competent to enter advanced NP practice? PMID- 16344716 TI - Pharmacogenetics of opioid receptors and addiction. AB - It is generally assumed that combinations of polymorphic alleles of different genes contribute to polygenetic disorders. Variants of the opioid receptors are the obvious candidates underlying addiction. Most research has focused on the coding variation A118G of the mu opioid receptor (MOPr), which replaces asparagine at position 40 by aspartate (Asn40Asp). However, to date, no conclusive evidence exists regarding which physiological effects this mutation may cause. Other rare polymorphisms in the MOPr cause marked effects (e.g. impairment of G-protein coupling) but, due to their low frequency, their role in addiction is questionable. A large number of polymorphic sites have been found in the putative promoter region of the mu opioid receptor. These variations may alter the expression level of the receptors in neurones, but the functional relevance of promoter polymorphisms is hard to predict. In the delta opioid receptor, surprisingly few polymorphisms have been detected within the coding region, and a clear association with addiction has not been shown to date. The kappa opioid receptor contains mainly silent polymorphisms. Nevertheless, there are studies reporting positive associations of silent mutations in the three opioid receptors with drug addiction. The underlying mechanism remains unclear, but examples exist indicating that silent mutations affect mRNA stability. Taken together, the coding polymorphisms, which are rather frequent, reveal no convincing association. The vast number of non-coding, intronic or promoter polymorphisms in the opioid receptors may influence addictive behaviour, but these polymorphisms are far less studied, and their physiological significance remains to be demonstrated. PMID- 16344717 TI - The Arg389Gly beta1-adrenoceptor polymorphism does not affect cardiac effects of exercise after parasympathetic inhibition by atropine. AB - In vitro, Arg389Gly beta1-adrenoceptor (AR) polymorphism exhibits decreased beta AR signalling. In vivo, beta1-AR-mediated cardiac effects of exercise showed no genotype-dependent differences in Arg389 vs. Gly389 beta1-AR subjects. We studied in 16 male subjects homozygous Arg389 or Gly389 beta1-AR, whether blockade of parasympathetic activity might unmask genotype-dependence of exercise effects. Subjects were infused with atropine (10 microg/kg i.v. loading dose followed by continuous i.v. infusion of 0.15 microg/kg/min throughout exercise-time); 20 min after start of atropine bicycle-exercise in supine position (25, 50, 75 and 100 W for 5 min each) was performed and heart rate, contractility, blood pressure, plasma noradrenaline and plasma-renin activity were assessed. Exercise-evoked increases in all but one parameters were not different between Arg389 and Gly389 beta1-AR subjects; only plasma noradrenaline increased slightly more in Gly389 vs. Arg389 beta1-AR subjects. IN CONCLUSION: It appears to be unlikely that lack of Arg389Gly beta1-AR genotype-dependence of exercise-effects can be explained by influences of parasympathetic activity. PMID- 16344718 TI - A study of TH01 and IGF2-INS-TH haplotypes in relation to smoking initiation in three independent surveys. AB - OBJECTIVES: Recent studies suggest an association between a microsatellite locus (TH01) located in intron 1 of the tyrosine hydroxylase gene (TH) and nicotine dependence. We aimed here to study whether both TH01 and haplotypes of the wider IGF2-INS-TH region influence initiation of regular smoking in current smokers. METHODS: A total of 3637 individuals from three independent studies (two of adults and one of adolescents) were analysed in relation to the age of first regular smoking (AFRS). Haplotypes and genotypes were obtained for the polymorphisms TH01, IGF2 ApaI, INS HphI and DRD4 VNTR (48 bp)n. Association between IGF2-INS-TH haplotypes and AFRS was tested by a regression model. A genotype-based genetic model assuming additivity was followed in order to estimate the effect of individual loci. RESULTS: Overall, no significant associations were found after correcting for multiple tests. However, an IGF2-INS TH haplotype (*5) was found to be nominally associated with AFRS at younger ages in adult smokers. Analyses of individual loci points to TH01 as a possible candidate influencing initiation of regular smoking. An AFRS-lowering trend nominally associated with allele 9 in a dosage-dependent manner was identified in both adult cohorts. TH01 did not show association or trend with age of initiation (first puff) either in adolescents or in the adolescents smoking regularly at age 18. CONCLUSION: This study adds to the genetic evaluation of the associations of TH01 with smoking predisposition. Differences between historical and prospective surveys, different biological pathways and possible functional roles of this microsatellite in smoking initiation are discussed. PMID- 16344719 TI - Association analysis of polymorphisms in serotonin 1B receptor (HTR1B) gene with heroin addiction: a comparison of molecular and statistically estimated haplotypes. AB - OBJECTIVES: 5-Hydroxytryptamine (serotonin)-1B receptors (HTR1B) may play an important role in psychiatric disorders and drug and alcohol dependence. In this study we report on genotype, molecular haplotype and statistically estimated haplotype analyses of previously identified polymorphisms in positions -261T>G, 161A>T, 129C>T, 861G>C and 1180A>G of the HTR1B gene in ethnically diverse populations (African-Americans, Caucasians, Hispanics and Asians) including 235 former heroin addicts and 161 control subjects from New York City. The objectives were to test for an association of molecular and statistically estimated haplotypes and genotypes in HTR1B gene with heroin addiction and to compare results provided by molecular and statistically estimated haplotyping methods. METHODS: Genotype analysis was performed using a standard TaqMan protocol. Molecular haplotype analysis of the subset of polymorphisms consisting of 261T>G, -161A>T and 129C>T was performed using a protocol specially designed by our group, using fluorescent PCR. This is based on use of allele-specific primers complementary to flanking polymorphisms and a fluorescently labeled sequence specific TaqMan probe set complementary to an internal polymorphism of the haplotype region. Every individual's statistically inferred haplotype pair agreed with the individual's haplotype pair determined by molecular haplotyping. RESULTS AND CONCLUSION: A point-wise significant association of haplotype pairs containing allele G at position 1180 with protective effect from heroin addiction in Caucasians was found. A point-wise nominally significant association of allele 1180G with a protective effect from heroin addiction was found in Caucasians. Statistically significant differences across four ethnic groups in control subjects for allelic frequencies of -261T>G and -161A>T were found. PMID- 16344720 TI - No association between C-reactive protein gene polymorphisms and decrease of C reactive protein serum concentration after infliximab treatment in Crohn's disease. AB - We recently showed an association between the FCGR3A V/F polymorphism and the biological response [assessed on the basis of a C-reactive protein (CRP) concentration decrease] to infliximab in Crohn's disease. The CRP and FCGR3A genes are located on the same 1q23 locus. The present study aimed: (i) to exclude a linkage disequilibrium (LD) between the two genes and (ii) to study the association between CRP polymorphisms and the response to infliximab, particularly the decrease in CRP after treatment, in Crohn's disease patients. FCGR3A (V/F) polymorphism and three CRP polymorphisms (-717G/A, 1444C/T, CRP 4A/G) were determined in 206 healthy blood donors and 189 Crohn's disease patients who had received infliximab for either refractory luminal or fistulizing Crohn's disease. Clinical response was defined as complete, partial or absent according to the same definition as in controlled trials. The biological response was defined on the basis of CRP decrease. There was no LD between CRP and FCGR3A in healthy donors or Crohn's disease patients. CRP polymorphisms had no impact on CRP decrease after infliximab. The proportions of Crohn's disease having a positive clinical or biological response were not statistically different among the various genotypes of CRP polymorphisms. There was no LD between CRP and FCGR3A polymorphisms. CRP polymorphisms were not associated with the response to infliximab in Crohn's disease. PMID- 16344721 TI - Protective effect of nonsteroidal anti-inflammatory drugs on colorectal adenomas is modified by a polymorphism in peroxisome proliferator-activated receptor delta. AB - OBJECTIVE: Nonsteroidal anti-inflammatory drugs (NSAIDs) are associated with a decreased risk of colorectal tumors. Single nucleotide polymorphisms (SNPs) in target genes of NSAID action, and their haplotypes, might modulate this protective effect. METHODS: A case-control study including 724 cases and 682 controls was used to evaluate the effect of NSAIDs on colorectal adenoma risk in The Netherlands, a country in which NSAID use is relatively low. Cases and controls were classified according to presence or absence of endoscopy-proven, pathology-confirmed colorectal adenomas, ever in their lives. Thirteen SNPs in four genes (PPARdelta, PPARgamma, PTGS1 and PTGS2) were genotyped in 787 subjects (384 cases and 403 controls). RESULTS: Compared to non-regular users (< 12 times/year), regular users of NSAIDs (> or = 12 times/year) had a lower risk of colorectal adenomas (odds ratio (OR): 0.75, 95% confidence interval (CI): 0.56 0.99). The results were similar for aspirin only. We found an interaction between SNP c.-789C>T in PPARdelta and NSAID use (P=0.03). The protective effect of NSAIDs was strengthened for regular users with the PPARdelta CT or TT genotypes (OR: 0.35, 95%CI: 0.11-1.13), whereas a positive association was observed for non regular users with these genotypes (OR: 2.24, 95%CI: 1.06-4.73) as compared to non-regular users with the CC genotype. Also, a statistically significant interaction between a major haplotype containing the minor allele of this SNP and NSAID use was observed. CONCLUSIONS: This study confirms the protective effect of NSAIDs and suggests a modulating effect of a SNP in the promoter of PPARdelta. PMID- 16344722 TI - CYP2E1 and catalase influence ethanol sensitivity in the central nervous system. AB - OBJECTIVES: Genetic factors are known to influence the sensitivity and tolerance to ethanol in humans and laboratory animals. Ethanol is metabolized to acetaldehyde mainly by the alcohol dehydrogenase pathway (ADHs) and, to a lesser extent, by microsomal oxidization (CYP2E1) and the catalase-H2O2 system. METHODS: In this study, we examined the role of CYP2E1 and catalase in ethanol metabolism and sensitivity, using transgenic knockout Cyp2e1(-/-) mice, acatalasemic (Cs/Cs) mice, double mutant Cyp2e1(-/-)/Cs/Cs mice and their respective wild-type counterparts 129/sv, C3H/HeJ, 129/sv X C3H/HeJ mice. Ethanol was administered to the mouse lines and ethanol pharmacokinetics and sleep times were evaluated. RESULTS: Although the rates of whole blood ethanol elimination following i.p. administration were found to be similar regardless of dose or genetic stock, Cs/Cs, Cyp2e1(-/-) and Cyp2e1(-/-)/Cs/Cs mice exhibited longer ethanol-induced sleep times, especially at higher ethanol doses. This infers that there is less acetaldehyde produced in the brains of these animals and is in opposition to the idea that increased acetaldehyde increases the actions of ethanol. The Cyp2e1(-/ ) animals produced lower whole blood levels of acetaldehyde than wild-type controls; however, this difference was seen only at higher doses of ethanol. The amount of acetaldehyde produced following the incubation of ethanol with liver and brain microsomes was greater in tissues derived from 129/sv than in those from Cyp2e1(-/-) mice. CONCLUSIONS: Although the contribution of CYP2E1 and catalase in ethanol oxidation may be of little significance, these enzymes appear to play a significant role in ethanol sensitivity in the brain. PMID- 16344723 TI - Genetic signature consistent with selection against the CYP3A4*1B allele in non African populations. AB - Cytochrome P450 3A enzymes (CYP3A) play a major role in the metabolism of steroid hormones, drugs and other chemicals, including many carcinogens. The individually variable CYP3A expression, which remains mostly unexplained, has been suggested to affect clinical phenotypes. We investigated the CYP3A locus in five ethnic groups. The degree of linkage disequilibrium (LD) differed among ethnic groups, but the most common alleles of the conserved LD regions were remarkably similar. Non-African haplotypes are few; for example, only four haplotypes account for 80% of common European Caucasian alleles. Large LD blocks of high frequencies were suggestive of selection. Accordingly, European Caucasian and Asian cohorts each contained a block of single nucleotide polymorphism (SNPs) with very high P excess values. The overlap between these blocks in these two groups contained only two of the investigated 26 SNPs and one of them was the CYP3A4*1B allele. The region centromeric of CYP3A4*1B exhibited high haplotype homozygosity in European Caucasians as opposed to African-Americans. CYP3A4*1B showed a moderate effect on CYP3A4 mRNA and protein expression, as well as on CYP3A activity assessed as Vmax of testosterone 6beta-hydroxylation in a liver bank. Our data are consistent with a functional relevance of CYP3A4*1B and with selection against this allele in non-African populations. The elimination of CYP3A4*1B involved different parts of the CYP3A locus in European Caucasians and Asians. Because CYP3A4 is involved in the vitamin D metabolism, rickets may have been the underlying selecting factor. PMID- 16344724 TI - Lack of somatic mutations in EGFR tyrosine kinase domain in hepatocellular and nasopharyngeal carcinoma. AB - Activating EGFR somatic mutations have been shown to predict treatment response to small molecules targeting the EGFR intracellular tyrosine kinase domain. Recent work on cell-lines and animal models had demonstrated an inhibitory effect of EGFR tyrosine kinase inhibitors in hepatocellular and nasopharyngeal carcinoma, and clinical trials in these tumour types are ongoing. There are few data on the presence or prevalence of EGFR mutations in hepatocellular and nasopharyngeal carcinomas. We studied exons 18-21 of the EGFR gene from 100 hepatocellular and 102 nasopharyngeal carcinomas, and found no exonic mutations of potential significance. Alternative mechanisms may be important for the observed activity of small molecule EGFR tyrosine kinase inhibitors in hepatocellular and nasopharyngeal carcinomas. PMID- 16344726 TI - Sonography for deep venous thrombosis: current and future applications. AB - Deep venous thrombosis (DVT) is a one of the most common problems facing the clinician in medicine today. It is often asymptomatic and goes undiagnosed with potentially fatal consequences. Ultrasound has become the "gold standard" in the diagnosis of deep venous thrombosis and with proper attention to technique sensitivity of this test is approximately 97%. An understanding of anatomy, pathophysiology, and risk factors is important. Thrombus formation usually begins beneath a valve leaflet below the knee. Approximately 40% will resolve spontaneously, 40% will become organized, and 20% will propagate. Whether or not a calf vein thrombus is identified, a repeat examination in 7 to 10 days is recommended in patients with risk factors or when deep venous thrombosis is suspected. The three main risk factors for thrombus formation are age greater than 75 years, previous history of deep venous thrombosis, and underlying malignancy. Other diagnostic studies include the contrast venogram, CT or MRI venogram, Tc99m Apcitide study, and the laboratory test D-Dimer. The D-Dimer study is being used more frequently as a screening test with 99% sensitivity in detecting thrombus, whether deep venous thrombosis or pulmonary embolism. However, specificity is only approximately 50% with many conditions leading to false-positive exams. Therefore, a negative examination is useful in avoiding other diagnostic studies, but a positive one may be misleading. Conditions that can lead to a false-positive examination include, but are not limited to diabetes, pregnancy, liver disease, heart conditions, recent surgery, and some gastrointestinal diseases. Like the sonogram, two negative D-Dimer studies a week apart exclude the diagnosis of deep venous thrombosis. Compression sonography with color Doppler remains the best overall test for deep venous thrombosis. It is easy to perform, less expensive than most "high tech" studies, can be performed as a portable examination, and is highly reliable when done properly. PMID- 16344727 TI - The role of ultrasound in renal insufficiency: the essentials. AB - Sonography is the best screening modality to evaluate patients presenting with renal insufficiency. Ultrasound findings can be normal in patients with renal disease, especially in prerenal azotemia and acute parenchymal renal disease. Echogenic kidneys indicate the presence of parenchymal renal disease; the kidneys may be of a normal size or enlarged. Small kidneys suggest advanced stage chronic kidney disease. Uncommonly, cystic disease of the kidney, especially adult type polycystic kidney disease may be the cause of the patient's renal insufficiency with bilaterally enlarged kidneys containing multiple cysts of various sizes. If hydronephrosis is present, the level and cause of the obstruction should be sought. When ultrasound cannot diagnose the level and cause of obstruction, other imaging modalities, including CT and MRI may be useful. When renovascular disease (arterial stenosis or venous thrombosis) is suspected, spectral and color Doppler can be useful in detecting abnormalities. PMID- 16344728 TI - Gestational trophoblastic disease: pictorial review. AB - Ultrasound is the modality of choice for evaluating normal or abnormal first trimester pregnancy. Sonography can usually provide a specific diagnosis in abnormal first trimester bleeding. When the sonographic appearance is correlated with the clinical presentation, accurate diagnosis is possible in most cases of gestational trophoblastic disease (GTD). Partial or complete hydatidiform moles can be diagnosed in early gestation. However certain cases will be missed if the curettage material is not sent for pathologic examination. Sometimes molar pregnancies have very unusual sonographic appearances. Sonography and Doppler imaging are helpful in diagnosing gestational trophoblastic disease, in determining whether invasive disease is present, in detecting recurrent disease, and in following the effectiveness of chemotherapy. This pictorial essay describes the pathogenesis, epidemiology, and sonographic spectrum of gestational trophoblastic disease. PMID- 16344729 TI - What Is YOUR Diagnosis?: Case 2. Takayasu Arteritis. PMID- 16344730 TI - Sonographic evaluation of the musculoskeletal soft tissue masses. AB - This article describes the sonographic appearance of a variety of soft tissue masses. We review the current indications for performing musculoskeletal sonography as a screening and diagnostic tool in the clinical management of soft tissue masses. PMID- 16344746 TI - Spleen and retroperitoneum: the essentials. AB - This is a survey of normal variants and pathologic conditions involving the spleen and retroperitoneum. The study focuses on the various sonographic appearances of trauma, infection, and neoplasm involving these areas in an attempt to complement works dealing specifically with the pancreas, kidneys, and great vessels. Ultrasound-guided intervention (biopsy, drainage) is included. PMID- 16344747 TI - Ultrasound imaging of fetal brain abnormalities: three essential anatomical levels. AB - Prenatal ultrasound evaluation of the fetal brain requires documentation of specific structures according to guidelines set by the American College of Radiology and the American Institute of Ultrasound in Medicine. Among these required structures are: cerebellum, cisterna magna, lateral cerebral ventricles, choroid plexus, midline falx, and cavum septum pellucidum. All these structures can be visualized in three crucial planes of imaging that include cisterna magna/cerebellum, cavum septum pellucidum, and ventricular atria. A systematic approach can be achieved by instituting a protocol that includes these three basic levels of imaging of the fetal head. These anatomical levels should be incorporated in daily routine for complete obstetric ultrasound evaluation. PMID- 16344748 TI - Ultrasound markers of fetal infection part 1: viral infections. AB - Diagnosis of fetal infection has depended on identification of pathogens by means of microbiological cultures, immunologic techniques, and special molecular biology techniques that can identify organisms known or suspected of being associated with adverse outcomes of pregnancy. Rubella, cytomegalovirus (CMV), herpes simplex virus (HSV), and human immunodeficiency virus (HIV), for example, are capable of gaining access to the amniotic cavity and producing fetal infection, even when amniotic membranes are intact. Intrauterine invasion by viruses can be associated with maternal symptoms of infection or can be completely silent. In many instances extensive fetal compromise with irreversible structural damage or fetal death will have occurred by the time infection is confirmed by culture or other histopathological methods. The evidence of fetal infection may be as subtle as nascent intrauterine growth restriction (IUGR), mildly inappropriate calcification of fetal organs, placenta, cord, and membranes, and failure to adequately develop fetal fat reserves. The evidence of infection may be as dramatic as obvious fetal malformation, severe central nervous system structural damage, or fetal death. Sonography is capable of detecting most of the grave alterations and some of the subtle effects that are typical of fetal infection. PMID- 16344751 TI - ["Uplavici: what's wrong"]. PMID- 16344752 TI - [Natal teeth: apropos of five cases]. AB - INTRODUCTION: The normal eruption of primary teeth begins with mandibular incisors about the age of 6 months. There are several qualifiers used to describe prematurely erupted teeth. Massler and Savara defined "natal teeth" as teeth present at birth and "neonatal teeth" as teeth erupted within the first month of life. The aim of this study, based on 5 cases, is to present clinical and structural characteristics, etiology, management techniques, complications and a review of the literature for natal teeth. MATERIAL AND METHODS: Our study is a retrospective study of 17,000 infants who were examined in the Neonatal Department of Children Hospital between 1984 and 2001. The material consisted of 5 infants with natal teeth, identified by the pediatrician. We analyzed the family history, the pregnancy history, the gender, the etiology, the complete examination of the infant, the clinical, the structural characteristics, the complications and the management of the teeth. RESULTS: We found 4 boys and 1 girl, 2 of the newborns were premature, all of them observed to have natal teeth. The incidence was 1: 3,400 births. There were 14 natal teeth, 10 incisors (70%) and 4 canines (30%), no molar was found. Nine of the teeth (6 incisors and 3 canines) were maxillary (65%) and 5 (4 incisors and 1 canine) were mandibular (35%). No morphological syndrome was discovered. Most of the teeth were mobile in all directions and were extracted because of the possibility of aspiration, the difficulty in feeding and the ulceration of the ventral surface of the tongue. DISCUSSION: According to the literature, this phenomenon is rare and the incisors are the teeth most commonly involved. Natal teeth are more common than neonatal teeth and nearly 90% of these teeth are the normal primary teeth. The presence of natal teeth is due to several factors related to an unknown cause of disturbed biological chronology. There is no conclusive evidence of a correlation between early eruption and systemic disorders, but some investigators suggest that natal teeth may be associated with certain syndromes. We must keep in mind that radiographic examination is essential for the differential diagnosis between supernumerary and normal primary teeth. The supernumerary teeth should always be extracted but the decision to extract a normal mature natal tooth should be done according to scientific knowledge, mobility of the tooth, local or general complications and parental opinion. PMID- 16344753 TI - [Anhidrotic ectomermal dysplasia (AED): four cases]. AB - INTRODUCTION: Anhidrotic ectodermal dysplasia (AED) or Christ-Siemens-Touraine syndrome is a rare, hereditary genodermatosis, classically X-linked recessive disorder. MATERIAL AND METHODS: [corrected] We report the cases of 3 children and a male adult. RESULTS: The mode of diagnosis, the clinical signs and the therapeutic option are detailed. DISCUSSION: AED is characterized by a malformative state derived from the ectodermal layer of the embryo which results in the triad: anhidrosis (or hypohidrosis), hypotrichosis, anodontia (or hypodontia). Hypohidrosis causes thermoregulation disorders, which in the infant, can be life threatening. It is important to recognize the affection early to avoid accidents of hyperthermia. Once the diagnosis is established, family investigations are necessary to determine whether it is a family form or a new sporadic case. Carrier mothers must be informed of the high risk recurrence for future male infants. Symptomatic maxillo-facial treatment strives to improve masticatory function and facial growth and thus limit the psychological impact and improve patient comfort. PMID- 16344754 TI - [Scalp surgery in children: principles and therapeutic aspects]. AB - Surgical treatment of the lesions of the scalp in children has specific objectives: excision of an extensive lesion using minimum operative procedures, hiding scar in the hair, correcting a wide and ungraceful scar. We reviewed our experience of 450 cases (1990-2000), and found that most common lesions in childhood were: sebaceous hamartoma, pigmentary naevus, post traumatic alopecia, aplasia cutis congenita and complicated hemangiomas. Different surgical procedures are discussed: excision, tissue expansion, hair grafts. Large excision can be performed before three months of life because of the good laxity of the scalp. Tissue expansion is particularly suited to the scalp. Aplasia cutis congenita is a special condition, with local vascular abnormalities: local flaps are prohibited. Hemangiomas may require early surgical treatment in the event of complications. PMID- 16344755 TI - [Transconjunctival inferior blepharoplasty]. AB - The transconjunctival approach is the appropriate standard for lower eyelid blepharoplasty in patients presenting with lower eyelid herniated fat without excess skin. A transconjunctival incision is made approximately 2 mm below the tarsal border and extended inferiorly following a plane posterior to the orbital septum. No conjunctival suture is necessary. Although transconjunctival blepharoplasty diminishes the occurrence of postoperative complications when compared to the transcutaneous method and constitutes a more comfortable option for the patient, we always recommend a preoperative test of the eyes. Patients must be informed of the risk of amaurosis. PMID- 16344756 TI - [Free fibular flaps in children: morbidity of the donor site and pitfalls to avoid]. AB - INTRODUCTION: We report a case of donor site morbidity late after harvesting a free fibular flap for reconstruction of the mandibular symphsis. CASE REPORT: An 8-year-old boy presented a valgus deformity of the ankle 4 years harvesting of a vascularized fibular graft for mandibular reconstruction. The deformation was repaired with a tibial bone slit and pin fixation. The biomechanical result was very satisfactory, allowing normal gait. DISCUSSION: We recall the rules for fibular flap harvesting needed to prevent deformation of the donor site in children aged less than 9 years. PMID- 16344757 TI - [Growth defects of fibular flaps in children]. AB - BACKGROUND: Mandibular reconstruction with fibula free flap is a challenge in pediatric patients because of the size of the bone and absence of growth without epiphyseal transplantation. CASE REPORT: We report 2 cases of free fibula flap in children for mandibular reconstruction. The 2 patients presented with growth insufficiency of the bone graft after few years of evolution. DISCUSSION: Growth insufficiency was treated with different surgical procedures which were discussed We review the consequences on growth after fibula free flap and the specific indications to achieve epiphyseal fibular growth in children. PMID- 16344758 TI - [Cellular neurothekoma in an 11-year-old child]. AB - INTRODUCTION: Neurothekoma is a rare benign tumor which must be distinguished from certain malignant tumors such as fibrohistiocyte tumors or plexiform cell tumors, neurotropic melanomas and clear-cell sarcoma. CASE REPORT: An 11-year-old girl consulted for a recurrent subcutaneous tumor of the chin which had been operated 4 months earlier. The resection was incomplete. A wider revision resection successfully stopped recurrence. The histology study established the diagnosis of neurothekoma due to the presence of mitosis atypia, cellular nodules, and extension to the hypodermis. Immunohistochemistry confirmed the diagnosis. DISCUSSION: Neurothekoma is a benign tumor observed in young women, mainly on the face. It occurs as a dermal cohesive mass without infiltration of the epidermis. The typical immunohistochemical pattern enables differential diagnosis with myxoid neurothekoma, melanocytic and nervous system tumors. Surgical resection is indicated. PMID- 16344759 TI - [Teratoid cyst: a rare tongue tumor in children]. AB - BACKGROUND: Dermoid cysts are rare benign tumours (only 20 cases have been reported) with 3 different histological types. Only 5 cases of teratoid cyst of the tongue have been described in the world literature, characterized by a respiratory cilied epithelium. OBSERVATION: We report the sixth case, in a one year-old boy with a macroglossia and permanent protrusion of the tongue. Ultrasonography showed a kystic, ovoid, homogeneous cyst. Treatment was a surgical resection. DISCUSSION: This case is similar to the other cases reported in the literature. PMID- 16344760 TI - [Management of a squamous-cell carcinoma T2N1 neoplasia on the free border of the tongue]. PMID- 16344761 TI - [Two radio-opaque lesions on the orthopantogram]. PMID- 16344828 TI - Contemporary issues in the field. PMID- 16344764 TI - [Angioneurotic edema and widsom tooth avulsion]. PMID- 16344829 TI - Generalized anxiety disorder: rethinking diagnosis and rating. PMID- 16344830 TI - Factors associated with the prodromal progression of schizophrenia that influence the course of the illness. AB - OBJECTIVE: This study examined factors influencing the prognosis of patients with different prodromal manifestations of schizophrenia and the association of diagnosis and antipsychotic treatment with the frequency of future acute inpatient care. METHODS: Data was collected from the medical records of 24 patients initially diagnosed with schizophrenia. RESULTS: Seventy-six percent of the patients (N=18) had at least one psychiatric assessment before the debut of schizophrenia. Patients who were assessed prior to the initial diagnosis of schizophrenia presented in two distinct time frames. Patients in the first wave received a different Axis I diagnosis, depending on the phase of prodrome upon time of initial evaluation. Receiving any Axis I diagnosis during the prodromal stage in wave one correlated with an increased need for acute inpatient treatment (P>.0001) in the following 3 years. Among the patients in wave two, those who received antipsychotic treatment required less acute inpatient care than those who did not (P=.004). CONCLUSION: Obtaining a psychiatric history of assessments and interventions during the prodromal period of schizophrenia can be useful for prognosis. PMID- 16344831 TI - The role of estrogen therapy in postpartum psychiatric disorders: an update. AB - OBJECTIVES: The aim of this review is to assess and summarize the existing literature data about the efficacy of estrogen replacement therapy (ERT) in preventing and treating postpartum psychiatric disorders. METHODS: An extensive and unrestrictive computerized search (from 1970 to June 2005) on MEDLINE/PubMed, TOXNET, EMBASE, and Cochrane Databases was conducted with the following search terms: estradiol, estrogen-treatment, hormonal treatment, therapy, postpartum depression, postnatal depression, puerperal depression, postpartum blues, postpartum psychosis, lactation, and breastfeeding. RESULTS: In assessing available literature information about the role of ERT in preventing and treating puerperal psychiatric diseases, all reviewed studies were found to suffer from severe methodological limitations. CONCLUSION: Well-designed and strictly focused multicenter trials are warranted in order to firmly establish the effectiveness of ERT in puerperal psychiatric disorders. PMID- 16344832 TI - Issues in the psychopharmacologic assessment and treatment of the orthodox Jewish patient. AB - As with members of other cultural and religious groups, patients within the Orthodox Jewish community present with their own distinct clinical psychiatric issues related to their unique beliefs and practices. This article reviews the existing literature and anecdotal experience on the psychopharmacologic assessment and treatment of Orthodox Jewish patients. Specific aspects examined include this group's perceived intense stigma in receiving treatment, the priority this community places on cognitive functioning, and how the influence of Jewish laws on marriage and sexual practices impacts one's treatment decisions. The relevance of Jewish dietary laws, the Sabbath, and the community's interest in alternative treatments are also discussed. The limited ethno psychopharmacology research related to Orthodox Jewish psychiatric patients is reviewed. We conclude that understanding issues such as these is critical if one is going to work within this cultural system in order to successfully address their mental health issues. However, the dearth of controlled research in this community needs to be addressed to provide more effective treatment. PMID- 16344833 TI - Brain stimulation techniques in the treatment of obsessive-compulsive disorder: current and future directions. AB - Recent studies on the epidemiology of obsessive-compulsive disorder (OCD) estimate 50 million patients suffer from OCD worldwide, thus making it a global problem. The treatment of OCD has changed substantially over the last 2 decades following the introduction of selective serotonin reuptake inhibitors, which provide symptom improvement in approximately 60% of patients. However, some patients remain resistant to the standard pharmacologic and behavioral treatments. Although some treatment-resistant patients respond to pharmacologic augmentations, others do not, and there is evidence that some of the most severe cases benefit from treatment with neurosurgical interventions. Besides pharmacologic, behavioral, and neurosurgical approaches, different brain stimulation methods-transcranial magnetic stimulation, deep brain stimulation, and electroconvulsive therapy-have been investigated in treatment-resistant patients with OCD. However, available data about the use of these techniques in OCD treatment are quite limited in terms of sample size and study design, given the difficulty in conducting standard blinded trials for these procedures. In addition, none of the mentioned treatments have received Food and Drug Administration approval for the treatment of OCD. Nevertheless, promising findings regarding efficacy, tolerability, and non-invasiveness and/or reversibility of these techniques have increased interest in investigating their use in treatment-resistant OCD. PMID- 16344834 TI - Magnetic resonance imaging changes in a patient with migraine attack and transient global amnesia after cardiac catheterization. AB - The presence of magnetic resonance imaging (MRI) abnormalities in patients with transient global amnesia has been an interesting phenomenologic finding. Several theories surround the occurrence of this syndrome, but little is known about its true physiopathology. We present a case of transient global amnesia after cardiac catheterization associated with migraine headache and MRI changes compatible with an ischemic insult. A discussion on potential explanations for this finding is made, as well as a review of the pertinent literature. PMID- 16344836 TI - Single- and multiple-dose pharmacokinetics of an oral mixed amphetamine salts extended-release formulation in adults. AB - OBJECTIVES: Assess the bioavailability of mixed amphetamine salts extended release (MAS XR) 30-mg capsules and the dose proportionality of pharmacokinetic measures for MAS XR 20, 40, and 60 mg. METHODS: Study A, an open-label single period study, and Study B, a randomized, open-label, three-way crossover study, were conducted in healthy adults in a clinical research unit. In Study A, 20 subjects received a single MAS XR 30-mg capsule by mouth daily for 7 days. In Study B, 12 subjects received single oral doses of MAS XR 20, 40, and 60 mg separated by 7-14-day washout periods. FINDINGS: Plasma dextroamphetamine (d amphetamine) and levoamphetamine (l-amphetamine) concentrations were measured using a validated LC-MS/MS method. In Study A, a 3:1 ratio of d-amphetamine to l amphetamine was observed for AUC0-inf and Cmax. Tmax was 4.2 and 4.3 hours for d amphetamine to l-amphetamine, respectively. In Study B, for d- and l-amphetamine, statistically significant differences were observed for AUC0-t, AUC0-inf, and Cmax between all doses; there was a linear relationship between pharmacokinetic variables and dose and Tmax was similar for each isomer (range: 4.5-5.3 hours) with all given MAS XR doses. CONCLUSION: The extent of exposure as assessed by mean AUC0-24 and Cmax reflected the 3:1 ratio of d-amphetamine to l-amphetamine in MAS XR 30-mg capsules. The pharmacokinetic profiles of MAS XR 20, 40, and 60 mg are dose proportional for the isomers. PMID- 16344837 TI - Long-term safety and effectiveness of mixed amphetamine salts extended release in adults with ADHD. AB - OBJECTIVE: Assess the long-term safety and effectiveness of mixed amphetamine salts extended release (MAS XR) in adults with attention-deficit/hyperactivity disorder (ADHD) combined subtype. METHODS: A 24-month, open-label extension of a 4-week, multicenter, double-blind, placebo-controlled, parallel-group, forced dose-escalation study of MAS XR in adults (>or=18 years of age) with ADHD. The 223 enrolled subjects started treatment at 20 mg/day for 1 week, with subsequent titration up to 60 mg/day for optimal therapeutic effects. At monthly visits, efficacy was assessed based on the ADHD Rating Scale IV (ADHD-RS-IV). Safety assessments included spontaneously reported adverse events, laboratory assessments, and monitoring of vital signs. FINDINGS: ADHD symptoms significantly improved for all subjects as measured by change from baseline in mean ADHD-RS-IV total scores (-7.2+/-13.04 unit points; P<.001); this was sustained for up to 24 months. The most common treatment-related adverse events were dry mouth (43% of subjects reporting at least one occurrence), infection (33%), insomnia (32%), anorexia/decreased appetite (32%), headache (30%), and nervousness (26%). Most adverse events were mild to moderate in intensity. CONCLUSION: Treatment with MAS XR 20-60 mg/day for adult ADHD was generally well tolerated and was associated with sustained symptomatic improvement for up to 24 months. PMID- 16344838 TI - An interim analysis of the Quality of Life, Effectiveness, Safety, and Tolerability (QU.E.S.T.) evaluation of mixed amphetamine salts extended release in adults with ADHD. AB - OBJECTIVE: Evaluate safety and efficacy of mixed amphetamine salts extended release (MAS XR) in adults with attention-deficit/hyperactivity disorder (ADHD). METHODS: 10-week interim analysis of the Quality of life, Effectiveness, Safety, and Tolerability (QU.E.S.T.) trial, an ongoing, 30-week, open-label, multicenter investigation of once-daily MAS XR (10-60 mg/day) in adults (>or=18 years of age) with ADHD in community practice settings. FINDINGS: With up to 10 weeks of open label MAS XR 10 to 60 mg/day (final visit mean dose: 37.2 mg/day), 725 adults exhibited rapid, sustained improvement in ADHD symptoms. At end point, significant decreases from baseline were seen in ADHD Rating Scale IV total scores (-19.8+/-11.6; P<.0001), hyperactivity/impulsivity subscale (-8.1+/-6.1; P<.0001), and inattentive subscale (-11.6+/-6.7; P<.0001). Most subjects (74.4%) were rated as very much/much improved. Based on the 36-item Short Form Health Survey (version 2), significant improvements in quality of life were seen in the domains of general health, physical and mental health, vitality, and social, emotional and physical role functioning (P<.0001). Few subjects (6.9%) withdrew due to adverse events; the most common MAS XR-related adverse events were decreased appetite and dry mouth (19.2% each), insomnia (17.8%), and headache (16.8%). CONCLUSION: In adults with ADHD, MAS XR treatment is generally safe and demonstrates significant improvement in ADHD symptoms and related quality of life. PMID- 16344839 TI - Long-term cardiovascular effects of mixed amphetamine salts extended release in adults with ADHD. AB - OBJECTIVE: To assess long-term cardiovascular effects of mixed amphetamine salts extended release (MAS XR) in adults with attention-deficit/hyperactivity disorder (ADHD) combined subtype. METHODS: 223 otherwise healthy adults (>or=18 years of age) with ADHD combined subtype were exposed to 480 msec (QTcF [corrected by Fridericia's formula] >454 msec). Seven subjects discontinued due to a cardiovascular adverse event (hypertension, n=5, palpitation/tachycardia, n=2); none of these events was reported as serious. Few subjects with normal baseline vital signs (using approved parameters at the time of study initiation) exhibited clinically significant abnormalities at end point; several subjects with borderline baseline values exhibited shifts to abnormal values during MAS XR therapy. CONCLUSION: Cardiovascular effects of long-term MAS XR ( or =85th percentile at age 2-3 years was a risk factor for subsequent asthma development in boys (hazard ratio (HR) 1.6 95% confidence interval (CI) 1.1, 2.4) but not girls (HR 0.8, 95% CI 0.5, 1.4). Similarly, boys with BMIs always > or =85th percentile were at increased risk for subsequent asthma development (HR 2.4, 95% CI 1.4, 4.4) but not girls (HR 1.5, 95% CI 0.7, 2.9). CONCLUSION: Boys with high body masses may be at an increased risk for developing asthma. PMID- 16344844 TI - The impact of childhood body mass index on timing of puberty, adult stature and obesity: a follow-up study based on adolescent anthropometry recorded at Christ's Hospital (1936-1964). AB - OBJECTIVE: To examine the impact of childhood body mass index (BMI) on timing of puberty, adult stature and obesity. DESIGN: A retrospective school-based cohort follow-up study. SUBJECTS: A total of 1520 men born between 1927 and 1956 with serial height and weight measurements from the age of 9 to 18, followed up in adulthood at a mean age of 63 years. MEASUREMENTS: Prepubertal (child) BMI; age at peak height velocity (APHV); adult leg length, waist and hip circumference (self-reported), and height and weight (measured). RESULTS: Boys with a higher childhood BMI tended to have an earlier puberty (per s.d. increase in child BMI: 0.31 years, 95% confidence interval (CI): -0.23 to -0.39, P < 0.001). Childhood BMI correlated strongly and positively with adult adiposity, as measured by BMI (1.24 kg/m(2), 95% CI: 1.04-1.45, P < 0.001), waist (1.94 cm, 95% CI: 1.50-2.37, P < 0.001) and hip (1.91 cm, 95% CI: 1.55-2.28, P < 0.001). It correlated inversely with leg length (-0.40 cm, 95% CI: -0.68 to -0.12, P = 0.005), but not significantly with height or trunk length. Boys with a later puberty tended to be taller (per s.d. increase in APHV: 0.57 cm, 95% CI: 0.17-0.97, P = 0.006) and less adipose as adults (-0.55 kg/m(2), 95% CI: -0.33 to -0.77, P < 0.001). CONCLUSION: The long-term consequences of childhood obesity into adulthood are well known, but the potential impact on early pubertal maturation is less well appreciated. The factors that contribute to the onset and persistence of childhood obesity require more public health focus. PMID- 16344845 TI - Crossvalidation of anthropometry against magnetic resonance imaging for the assessment of visceral and subcutaneous adipose tissue in children. AB - BACKGROUND: The study of the relationship between anthropometry and visceral adipose tissue (VAT) is of great interest because VAT is associated with many risk factors for noncommunicable diseases and anthropometry is easy to perform in clinical practice. The studies hitherto available for children have, however, been performed on small sample sizes. DESIGN: Pooling of the data of studies published from 1992 to 2004 as indexed on Medline. AIMS: To assess the relationship between anthropometry and VAT and subcutaneous adipose tissue (SAT) as measured by magnetic resonance imaging (MRI) in children and to analyze the effect of age, gender, pubertal status and ethnicity. SUBJECTS AND METHODS: Eligible subjects were 7-16 year-old, with availability of VAT and SAT, gender, ethnicity, body mass index (BMI) and waist circumference (WC). A total of 497 subjects were collected from seven different investigators and 407 of them (178 Caucasians and 229 Hispanics) were analyzed. RESULTS: Despite ethnic differences in MRI data, BMI, WC and age, no difference in VAT was found between Caucasians and Hispanics after correction for SAT and BMI. Univariate regression analysis identified WC as the best single predictor of VAT (64.8% of variance) and BMI of SAT (88.9% of variance). The contribution of ethnicity and gender to the unexplained variance of the VAT-WC relationship was low (< or =3%) but significant (P < or =0.002). The different laboratories explained a low (< or =4.8%) but significant (P < 0.0001) portion of the unexplained variance of the VAT-WC and SAT-BMI relationships. Prediction equations for VAT (VAT (cm(2)) = 1.1 x WC (cm)-52.9) and SAT (SAT (cm(2)) = 23.2 x BMI (kg/m(2))-329) were developed on a randomly chosen half of the population and crossvalidated in the remaining half. The pure error of the estimate was 13 cm(2) for VAT and 57 cm(2) for SAT. CONCLUSIONS: WC can be considered a good predictor of VAT as well as BMI of SAT. The importance of ethnicity and gender on VAT estimation is not negligible. PMID- 16344846 TI - Cardiovascular fitness and physical activity in children with and without impaired glucose tolerance. AB - OBJECTIVE: To examine differences in cardiovascular fitness (VO(2max)) and physical activity levels in overweight Hispanic children with normal glucose tolerance (NGT) vs impaired glucose tolerance (IGT). PARTICIPANTS: A total of 173 overweight (BMI percentile 97.0 +/- 3.1) Hispanic children ages 8-13 years with a family history of type 2 diabetes. METHODS: VO(2max) was measured via a maximal effort treadmill test and open circuit spirometry. Physical activity was determined by questionnaire. Glucose tolerance was established by a 2-h oral glucose challenge (1.75 g of glucose/kg body weight). IGT was defined from an oral glucose tolerance test as a 2-h plasma glucose level > or =140 and <200 mg/dl. RESULTS: IGT was detected in 46 of the 173 participants (approximately 27%); no cases of type 2 diabetes were identified. No significant differences were found between youth with NGT and those with IGT in absolute VO(2max) (2.2 +/ 0.6 vs 2.1 +/- 0.5 l/min), VO(2max) adjusted for gender, age, and body composition (2.2 +/- 0.2 vs 2.1 +/- 0.2 l/min), or recreational physical activity levels (8.7 +/- 8.2 vs 6.9 +/- 6.2 h/week). CONCLUSION: Overweight Hispanic youth with IGT exhibit similar levels of VO(2max) and physical activity compared to their NGT counterparts. Longitudinal analyses are necessary to determine whether fitness/activity measures contribute significantly to diabetes risk over time in this group. PMID- 16344847 TI - The link between obesity and prostate cancer: the leptin pathway and therapeutic perspectives. AB - Obesity-associated prostate cancer (PCa) remains controversial, although most studies rely on body mass index evaluation, which is an indirect measure of fatness. Studies using body fat measurement and disease stratification according to PCa stage found stronger associations between obesity and PCa. Leptin is a pleiotrophic hormone mainly synthesized by adipocytes that acts in peripheral organs such as the prostate. This article reviews obesity-associated leptin's pathophysiological role in PCa progression. PCa development results from some known risk factors. Currently, there is enough evidence suggesting that leptin is an additional factor involved in advanced PCa occurrence, and obesity association with high-grade disease. Life-long exposure to genetic and/or environmental susceptibility factors that predispose to obesity and higher leptin levels may increase the risk for advanced PCa. PMID- 16344848 TI - Lower extremity skeletal muscle function in persons with incomplete spinal cord injury. AB - STUDY DESIGN: A cross-sectional study design. OBJECTIVES: To characterize and specifically quantify impairments in muscle function after chronic incomplete spinal cord injury (SCI). SETTING: University of Florida, Gainesville, FL, USA. METHODS: Voluntary and electrically elicited contractile measurements were performed and voluntary activation deficits were quantitatively determined in the knee extensor and ankle plantar flexor muscle groups in 10 individuals with chronic incomplete SCI (C5-T8, ASIA C or D) and age-, gender-, height- and body weight matched healthy controls. RESULTS: Persons with incomplete-SCI were able to produce only 36 and 24% of the knee extensor torque and 38 and 26% of the plantar flexor torque generated by noninjured controls in the self-reported less involved and more-involved limbs, respectively (P<0.05). In addition, both indices of explosive or instantaneous muscle strength, torque200 (absolute torque reached at 200 ms) and the average rate of torque development (ARTD) were dramatically reduced in the ankle plantar flexor and knee extensor muscle groups in persons with incomplete-SCI. However, the deficit in instantaneous muscle strength was most pronounced in the ankle plantar flexor muscles, with an 11.7 fold difference between the torque200 measured in the self-reported more involved limb and a 5-fold difference in the less-involved limb compared to control muscles. Voluntary activation deficits ranged between 42 and 66% in both muscle groups. Interestingly, electrically elicited contractile properties did not differ between the groups. CONCLUSION: The resultant impact of incomplete-SCI is that affected muscles not only become weak, but slow to develop voluntary torque. We speculate that the large deficit in torque200 and ARTD in the ankle plantar flexors muscles of persons with incomplete-SCI may limit locomotor function. The results presented in this study provide a quantitative and sensitive assessment of muscle function upon which future research examining rehabilitation programs aimed at restoring muscle function and promoting functional recovery after incomplete-SCI may be based. PMID- 16344849 TI - Cervical myelopathy induced by pseudogout in ligamentum flavum and retro-odontoid mass: a case report. AB - STUDY DESIGN: Retrospective case study. OBJECTIVES: Report a rare case of cervical myelopathy induced by calcium pyrophosphate dehydrate (CPPD) deposition in multiple cervical levels. SETTING: An area teaching hospital in Taiwan. METHOD: A patient with cervical myelopathy was evaluated by computerized tomography (CT) scan and magnetic resonance (MR) image. CPPD deposition known as pseudogout was diagnosed and approved by a polarized microscope. RESULT: A prominent hypertrophy of ligmentum flavum and a retro-odontoid bulging mass induced cord compression were found in CT scan and MR image. CPPD deposition was confirmed by the histological examinations in the ligamentum flavum at the spinal levels of C3-C6. After decompression surgery of spine and comprehensive rehabilitation, the patient's neurological symptoms subsided and her neurological functions improved leading to a good prognosis. CONCLUSION: CPPD deposition in cervical spine occurring at multiple levels is rare. Image studies with CT scan and MR are complementary in the diagnosis of CPPD-induced myelopathy. Surgical decompression is always required and expected to have a good outcome. PMID- 16344850 TI - Neurogenic bowel dysfunction score. AB - STUDY DESIGN: Cross-sectional questionnaire study. OBJECTIVES: To develop and validate a symptom-based score for neurogenic bowel dysfunction (NBD): NBD score. SETTING: University Hospital of Aarhus, Denmark. METHODS: A questionnaire including questions about background parameters (n=8), faecal incontinence (n=10), constipation (n=10), obstructed defecation (n=8), and impact on quality of life (QOL) (n=3) was sent to 589 Danish spinal cord injured (SCI) patients. The reproducibility and validity of each item was tested in 20 and 18 patients, respectively. Associations between items and impact on QOL were determined by logistic regression analysis. The NBD score was constructed from items with acceptable reproducibility and validity that were significantly associated with impact on QOL. Based on odds ratios for associations between items and impact on QOL, each item was given a corresponding number of points in the NBD score. RESULTS: A total of 424 SCI patients responded. The following 10 items met the criteria above: frequency of bowel movements (0-6 points), headache, perspiration or discomfort before or during defecation (0-2 points), tablets and drops against constipation (0-2 points each), time used for each defecation (0-7 points), frequency of digital stimulation or evacuation (0-6 points), frequency of faecal incontinence (0-13 points), medication against faecal incontinence (0-4 points), flatus incontinence (0-2 points) and perianal skin problems (0-3 points). Differences in NBD score among patients reporting no, little, some or major impact on QOL were statistically significant (all P<0.001). CONCLUSION: Based on valid and reproducible questions, we have constructed a score for NBD that is correlated to impact on QOL. PMID- 16344851 TI - Study of the effectiveness of bronchial clearance in subjects with upper spinal cord injuries: examination of a rehabilitation programme involving mechanical insufflation and exsufflation. AB - STUDY DESIGN: Randomized controlled trial. OBJECTIVES: To establish whether the use of mechanical insufflation/exsufflation leads to a significant improve in tracheostomy subjects with upper spinal cord injuries (C1-C7), ASIA classification grade A and bronchial hyper secretion the following parameters: forced vital capacity (FVC), forced exspiratory volume on the first second (FEV1), FEV1/FVC, peak exspiratory flow (PEF), arterious pressure of O(2) (Pa O(2)), arterious pressure of CO(2) (Pa CO(2)), pH, saturation of O(2) (Sa O(2)). SETTING: Spinal Cord Unit, Montecatone Rehabilitation Institute, Imola, Italy. METHODS: The patients were split into two groups: the experimental group (EG) and the control group (CG). Each patient was given 10 treatments: the EG was given manual respiratory kinesitherapy associated with mechanical insufflation/exsufflation with inhale and exhale pressure between 15 cm H(2)O and 45 cm H(2)O, while the CG was only given manual kinesitherapy. The treatment sessions covered the necessary amount of time in order to achieve sufficient clearance. The study has been approved by the local ethic committee. The patients were informed before being enrolled in that study and gave their written consent. RESULTS: At the end of the treatment associated with the mechanical insufflation/exsufflation, the EG showed a significant increase in FVC, FEV1 and PEF, although it was not possible to compare the latter with the CG. The other parameters were also subject to variations, although not statistically significant. The CG did not show signs of any significant change. CONCLUSION: The use of mechanical insufflation/exsufflation in subjects with the aforementioned characteristics is shown to be an effective adjunct to manual respiratory kinesitherapy, since it makes it possible to achieve adequate bronco-pulmonary clearance, even removing thick, deep secretions and making it possible to insufflate any areas affected by atelectasis. PMID- 16344853 TI - Human dignity: a guide to policy making in the biotechnology era? AB - This article explores the ways in which human dignity is used in debates about controversial biotechnologies, including biobanks, human gene patents, stem cell research and human cloning. Increasingly, human dignity is used as a form of general condemnation and as blanket justification for regulatory restraint. However, this use of human dignity marks a significant departure from the traditional, human-rights informed view of human dignity that has dominated bioethics debates for decades. In addition, on its own, it stands as dubious justification for policies that are aimed at constraining controversial biotechnologies. PMID- 16344852 TI - Use of c-fos to identify activity-dependent spinal neurons after stepping in intact adult rats. AB - STUDY DESIGN: An investigation of c-fos activation pattern in spinal neurons of intact adult rats after acute bouts of treadmill locomotion. OBJECTIVES: To map spinal neurons that are involved in quadrupedal treadmill stepping of intact adult rats by using c-fos as a marker. SETTINGS: Los Angeles, CA, USA. METHODS: Spinal cord sections of rats that were not stepped (n = 4) were used to map the FOS-positive (+) neurons under basal conditions. The stepped group (n = 16) was placed on a treadmill to step quadrupedally for varying durations to induce c-fos activity. Spinal cord sections of thoracic and lumbar segments of Stp and Nstp rats were processed using a c-fos antibody, choline acetyl transferase and heat shock protein 27 for identifying motoneurons. RESULTS: Stepping induced a greater number of FOS+ neurons than was observed in rats that did not step on the treadmill. There was a rostrocaudal and a dorsoventral gradient of FOS labeled neurons. The number of FOS+ neurons increased with the duration of treadmill stepping. Significant increases in FOS+ neurons were in the most medial parts of laminae IV, V, and VII. FOS+ motoneurons increased with treadmill stepping, particularly in large motoneurons (> or = 700 microm2). CONCLUSION: These data suggest that FOS can be used to identify activity-dependent neuronal pathways in the spinal cord that are associated with treadmill stepping, specifically in lamina VII and in alpha motoneurons. SPONSORSHIP: NIH NS16333, NS40917, and the Christopher Reeve Paralysis Foundation (CRPF VEC 2002). PMID- 16344854 TI - Influence of mirtazapine on plasma concentrations of neuroactive steroids in major depression and on 3alpha-hydroxysteroid dehydrogenase activity. AB - Concentrations of 3alpha-reduced neuroactive steroids are altered in depression and normalize after antidepressant pharmacotherapy with selective serotonin re uptake inhibitors (SSRIs). We investigated the impact of mirtazapine on the activity of a key neurosteroidogenic enzyme, the 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD), and on the levels of neuroactive steroids in relation to clinical response. A total of 23 drug-free in-patients suffering from a major depressive episode (DSM-IV criteria) underwent 5-week treatment with mirtazapine (45 mg/day). Plasma samples were taken weekly at 0800 and quantified for neuroactive steroids by means of combined gas chromatography/mass spectrometry analysis. Enzyme activity was determined by assessment of steroid conversion rates. Irrespective of clinical outcome, there were significant increases in 3alpha,5alpha-tetrahydroprogesterone, 3alpha,5beta-tetrahydroprogesterone, 5alpha dihydroprogesterone, and 5beta-dihydroprogesterone after mirtazapine treatment, whereas 3beta,5alpha-tetrahydroprogesterone levels were significantly decreased. In vitro investigations demonstrated a dose-dependent inhibitory effect of mirtazapine on the activity of the microsomal 3alpha-HSD in the oxidative direction (conversion of 3alpha,5alpha-tetrahydroprogesterone to 5alpha dihydroprogesterone). Mirtazapine affects neuroactive steroid composition similarly as do SSRIs. The inhibition of the oxidative pathway catalyzed by the microsomal 3alpha-HSD is compatible with an enhanced formation of 3alpha-reduced neuroactive steroids. However, the changes in neuroactive steroid concentrations more likely reflect direct pharmacological effects of this antidepressant rather than clinical improvement in general. PMID- 16344855 TI - Nonoverlapping expression patterns of ALK1 and ALK5 reveal distinct roles of each receptor in vascular development. AB - Transforming growth factor beta (TGF-beta) transmits signals through a heterotetrameric cell-surface complex of type II (TGFBR2) and type I (activin receptor-like kinase 5, ALK5; TGFBR1) serine/threonine kinase receptors, as well as Smad2/3. We have previously shown that another type I receptor, ALK1 (ACVRL1), can also mediate TGF-beta signals via BMP-activated Smads in vascular endothelial cells (ECs). Our group and others have proposed the hypothesis that two TGF-beta signaling pathways via ALK1 and ALK5 in vascular ECs may play a balancing role for controlling the proliferation and migration of ECs during angiogenesis. To address in vivo roles of this balance in vascular development, we have created a knockin mouse line that carries a lacZ reporter in the Alk5 gene locus (Alk5(lacZ)). Throughout development, a well-defined, nonubiquitous expression pattern of Alk5 expression was observed in multiple tissues, and organs. Overall, a high level of Alk5 expression was found in perichondria, periostea, and the mesenchymal layers underlying epithelia in the kidney, lung, and gallbladder. In blood vessels, contrasting to predominant Alk1 expression in arterial endothelium, Alk5 expression was localized in the medial and adventitial layers of blood vessels, but was undetectable in the intimal layer. In addition, although Alk5-null embryos exhibit a defect in the formation of vascular smooth muscle layers, the lumens of blood vessels are generated properly, which stands in contrast to the severe dilation of the vascular lumens in Alk1-null mice. These mutually exclusive expression patterns of Alk1 and Alk5 in blood vessels, as well as the undisturbed formation of the vascular lumens in Alk5-null embryos, suggest that each type I receptor has its own unique functions in vascular development. The Alk5(lacZ) mice will be a valuable resource in identifying the in vivo cellular targets of TGF-beta family signals mediated by Alk5, both during embryonic development as well as in diverse pathological conditions. PMID- 16344856 TI - Exploration of global gene expression in human liver steatosis by high-density oligonucleotide microarray. AB - Understanding the molecular mechanisms underlying fatty liver disease (FLD) in humans is of major importance. We used high-density oligonucleotide microarrays (22.3 K) to assess the mechanisms responsible for the development of human liver steatosis. We compared global gene expression in normal (n=9) and steatotic (n=9) livers without histological signs of inflammation or fibrosis. A total of 34 additional human samples including normal (n=11), steatosis (n=11), HCV-related steatosis (n=4) or steatohepatitis associated with alcohol consumption (n=4) or obesity (n=4) were used for immunohistochemistry or quantitative real-time PCR studies. With unsupervised classification (no gene selection), all steatotic liver samples clustered together. Using step-down maxT multiple testing procedure for controlling the Family-Wise Error-Rate at level 5%, 110 cDNAs (100 over- and 10 underexpressed) were found to be differentially expressed in steatotic and normal livers. Of them were genes involved in mitochondrial phosphorylative and oxidative metabolism. The mean ratio of mitochondrial DNA to nuclear DNA content was higher in liver steatosis compared to normal liver biopsies (1.12+/-0.14 vs 0.67+/-0.10; P=0.01). An increased expression of genes involved in inflammation (IL-1R family, TGFB) was also observed and confirmed by quantitative RT-PCR or immunochemistry. In steatohepatitis, an increase of the protein expression of mitochondrial antigens, IL-1R1, IGF2 and TGFB1 was also observed, interleukin 1 receptor being always strongly expressed in steatohepatitis linked to alcohol or obesity. In conclusion, mitochondrial alterations play a major role in the development of steatosis per se. Activation of inflammatory pathways is present at a very early stage of steatosis, even if no morphological sign of inflammation is observed. PMID- 16344857 TI - Tauroursodeoxycholic acid inserts the bile salt export pump into canalicular membranes of cholestatic rat liver. AB - Ursodeoxycholic acid exerts anticholestatic effects in chronic cholestatic liver disease in humans as well as in experimental animal models of cholestasis. Its taurine conjugate, TUDCA, was recently shown to stimulate insertion of the apical conjugate export pump, Mrp2 (ABCC2), into canalicular membranes of rat hepatocytes made cholestatic by exposure to taurolithocholic acid (TLCA). The aim of this immunoelectronmicroscopic study was to test whether TLCA and TUDCA modulate the canalicular density of the other key apical transporter, the bile salt export pump, Bsep (ABCB11), in a similar way. Immunoelectronmicroscopic analysis of Bsep density on canalicular membranes, microvilli, and pericanalicular area of hepatocytes was performed in rat liver tissue prepared after liver perfusion with bile acids or carrier medium only. TLCA (10 micromol/l for 50 min) decreased Bsep density in canalicular membranes to 31% of controls (P<0.05) when bile flow was reduced to 35% of controls (P<0.05). Concomitantly, Bsep density in a 1 microm pericanalicular zone increased to 202% (P<0.05) indicating effective retrieval of Bsep from the canalicular membrane induced by TLCA. Coadministration of TUDCA (25 micromol/l) led to a 3.2-fold increase of Bsep density in canalicular membranes equal to control liver (P<0.05 vs TLCA) in association with a 3.8-fold increase of bile flow (P<0.05 vs TLCA). Stimulation of apical membrane insertion of key transporters like the bile salt export pump, Bsep, and-as previously shown-the conjugate export pump, Mrp2, may contribute to the anticholestatic action of UDCA amides in cholestatic conditions. PMID- 16344858 TI - Infusing active learning into the large-enrollment biology class: seven strategies, from the simple to complex. PMID- 16344859 TI - From the National Academies: ongoing challenges to evolution education: resources and activities of the National Academies. PMID- 16344860 TI - Video views and reviews: gastrulation and the fashioning of animal embryos. PMID- 16344863 TI - SAMJ--Africa's top open access medical journal. PMID- 16344862 TI - The beta-glucan receptor dectin-1 recognizes specific morphologies of Aspergillus fumigatus. AB - Alveolar macrophages represent a first-line innate host defense mechanism for clearing inhaled Aspergillus fumigatus from the lungs, yet contradictory data exist as to which alveolar macrophage recognition receptor is critical for innate immunity to A. fumigatus. Acknowledging that the A. fumigatus cell wall contains a high beta-1,3-glucan content, we questioned whether the beta-glucan receptor dectin-1 played a role in this recognition process. Monoclonal antibody, soluble receptor, and competitive carbohydrate blockage indicated that the alveolar macrophage inflammatory response, specifically the production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1alpha (IL-1alpha), IL-1beta, IL-6, CXCL2/macrophage inflammatory protein-2 (MIP-2), CCL3/macrophage inflammatory protein-1alpha (MIP-1alpha), granulocyte-colony stimulating factor (G-CSF), and granulocyte monocyte-CSF (GM-CSF), to live A. fumigatus was dependent on recognition via the beta-glucan receptor dectin-1. The inflammatory response was triggered at the highest level by A. fumigatus swollen conidia and early germlings and correlated to the levels of surface-exposed beta glucans, indicating that dectin-1 preferentially recognizes specific morphological forms of A. fumigatus. Intratracheal administration of A. fumigatus conidia to mice in the presence of a soluble dectin-Fc fusion protein reduced both lung proinflammatory cytokine/chemokine levels and cellular recruitment while modestly increasing the A. fumigatus fungal burden, illustrating the importance of beta glucan-initiated dectin-1 signaling in defense against this pathogen. Collectively, these data show that dectin-1 is centrally required for the generation of alveolar macrophage proinflammatory responses to A. fumigatus and to our knowledge provides the first in vivo evidence for the role of dectin-1 in fungal innate defense. PMID- 16344864 TI - Better ultrasound service, less misguided litigation. PMID- 16344865 TI - Antiretroviral treatment in the Northern Cape. PMID- 16344866 TI - Zebras and bergkwaggas--the case for research on rare and very rare genetic diseases in South Africa. PMID- 16344867 TI - The "worried-well", insulin resistance and metformin therapy. PMID- 16344868 TI - Legal, but is it right? PMID- 16344869 TI - SAMA and sexuality--breaking the silence. PMID- 16344870 TI - The plague of socialism. PMID- 16344871 TI - African traditional healers. PMID- 16344873 TI - Societal consequences of Noah's alcohol misuse. PMID- 16344874 TI - World first for South African TB treatment guidelines. PMID- 16344875 TI - The dangers of going without. PMID- 16344876 TI - Built for the rich, serving the poor. PMID- 16344877 TI - Training and development needs of medical practitioners involved in the medico legal examination of survivors of sexual assault. PMID- 16344878 TI - Oxygen therapy and wound healing--topical oxygen is not hyperbaric oxygen therapy. PMID- 16344879 TI - An unusual complication of intestinal amoebiasis. PMID- 16344880 TI - Supporting the sexual and reproductive rights of HIV-infected individuals. PMID- 16344881 TI - Cot death controversies. PMID- 16344882 TI - Lipohypertrophy--a complication of insulin injections. PMID- 16344883 TI - Intravenous glucose preparation as the source of an outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae infections in the neonatal unit of a regional hospital in KwaZulu-Natal. AB - In the last week of May 2005, staff at Mahatma Gandhi Memorial Hospital in KwaZulu-Natal realised that many babies in the high-care nursery ward had bloodstream infections involving Klebsiella pneumoniae bacteria. Attempts to identify a common source of infection failed. The ward was therefore closed and new babies needing high care were admitted to another empty ward. Despite this, babies still became infected. This resulted in a request for assistance from the Department of Medical Microbiology of the Nelson R Mandela School of Medicine. A search for common factors through case history studies of the 26 infected babies showed that blood cultures of the babies remained positive despite the administration of appropriate antibiotics. Different options that could explain this were investigated. The organism was found in intravenous glucose preparations used for multiple dosing. Unopened vials of the same medication were sterile. The nursery was found to lack proper hand-wash facilities and to be overcrowded and understaffed. Reinforcement of hand hygiene and a ban on the multiple dosing of medicines stopped the outbreak. In conclusion, this outbreak resulted from a combination of factors among which lack of hand hygiene and multiple dosing of an intravenous glucose preparation were most significant. PMID- 16344884 TI - Antibiotic use and resistance in public-sector hospitals in KwaZulu-Natal. AB - OBJECTIVE: To investigate a possible association between level of care, antibiotic use and antimicrobial resistance in 16 public-sector hospitals providing different levels of care in KwaZulu-Natal. DESIGN: A multicentre surveillance study was undertaken in 16 hospitals at three progressive levels of health care (district, regional, tertiary) where each hospital submitted 100 consecutive, non-repetitive isolates judged in the laboratory to be of potential clinical significance. Isolates were identified and susceptibility testing was undertaken using the Kirby-Bauer disc diffusion method with minimum inhibitory concentrations (MICs) extrapolated on an automated reading system. Isolates were grouped according to their natural resistance profiles, and percentage susceptibility, mean percentage susceptibility and standard deviation to each antibiotic were stratified within and across hospital levels. Antibiotic use data were expressed as the number of daily divided doses (DDDs) per 1000 patient-days. SETTING: Two tertiary, 9 regional and 5 district public hospitals in KwaZulu Natal. OUTCOME MEASURES: Percentage susceptibility. RESULTS: There was a general trend among the 1 270 isolates of highest susceptibility in district hospitals, followed by regional and then tertiary hospitals. This is consistent with the referral system where health conditions become increasingly severe/complex requiring greater antibiotic use and broader-spectrum agents at progressive hospital levels, with statistical significance (p < 0.05) evident where sample numbers were relatively large. Trend variations could be associated with the qualitative and quantitative differences in antibiotic use, albeit without statistical corroboration. Three per cent of the total number of isolates were sensitive to all antibiotics tested and 6% were resistant to a single agent only. The remaining 91% showed acquired resistance to more than one drug. The standard deviation ranged from 0% to 55%. CONCLUSIONS: This study showed that resistance profiles among bacteria varied greatly within and across hospital levels. While antibiotic use varied as much, a statistically significant correlation between use and resistance could not be established. It was therefore postulated that the effect of selection pressure was obscured by other resistance determinants apparent in public hospitals in resource-poor settings. On a clinical level, the study showed that resistance profiles among bacteria vary too much to allow a national antibiotic policy as proposed in the standard treatment guidelines. Rather, such guidelines should be directed to specific profiles found in different hospitals and at different levels of health care. Regular surveillance to adjust such guidelines is essential. PMID- 16344885 TI - Impact of DDT re-introduction on malaria transmission in KwaZulu-Natal. AB - OBJECTIVES: To determine whether the re-introduction of DDT in KwaZulu-Natal had any effects on malaria transmission in the province. DESIGN, SETTING AND SUBJECTS: The 2000 malaria epidemic in KwaZulu-Natal has been attributed to pyrethroid-resistant anopheles mosquitoes in the area. Previous studies have shown that these mosquitoes are still susceptible to DDT. To determine whether DDT re-introduction had any impact on malaria transmission in KwaZulu-Natal, the following variables (pre- and post-epidemic) were investigated: (i) the number of reported cases; and (ii) the distribution of Anopheles funestus in relation to the insecticides sprayed. OUTCOME MEASURES: The notified malaria cases and the distribution of A. funestus were measured to determine the effects of DDT re introduction on malaria transmission. RESULTS AND CONCLUSION: After DDT re introduction, the number of malaria cases decreased to levels lower than those recorded before the epidemic. A. funestus appears to have been eradicated from the province. The combination of an effective insecticide and effective antimalarial drugs in KwaZulu-Natal has resulted in a 91% decline in the malaria incidence rate. Unfortunately the continued exclusive use of DDT within the malarious areas of the province is threatened by the emergence of insecticide resistance. PMID- 16344886 TI - Medico-legal aspects of pathology--current dilemmas regarding confidentiality and disclosure. PMID- 16344887 TI - Body fat distribution and insulin resistance. PMID- 16344888 TI - Acne guideline 2005 update. AB - OBJECTIVE: The guidelines on the management of acne vulgaris have been developed in an attempt to improve the outcomes of acne treatment in South Africa. This extremely common condition has a major impact on the quality of life of South African young people and it is expected that if implemented, these guidelines will play a role in improving this situation. RECOMMENDATIONS: All health care workers involved in the management of acne should take note of these guidelines and endeavour to implement them in clinical practice. All treatment methods and procedures not substantiated by evidence from the literature should be discontinued and avoided to decrease the financial burden of acne treatment. VALIDATION: These guidelines were developed through general consensus by a group of about 40 internationally recognised experts in the field of acne treatment (the Global Alliance to Improve Outcomes in Acne, see details below) and further refined for South African circumstances by the majority of South African dermatologists who attended a series of six discussions held in the major centres of South Africa during 2004. GUIDELINE SPONSOR: The meetings of the Global Alliance to Improve Outcomes in Acne as well as the South African discussion meetings were sponsored by Galderma. PMID- 16344889 TI - Progress in the study of vagal control of cardiac ventricles. AB - Autonomic nervous system plays an important role in the regulation of mammalian heart, and it is divided into the sympathetic and parasympathetic (vagal) subsystems. The parasympathetic (vagal) control of the atria involves modulation of chronotropic, dromotropic and inotropic activities, but the role of the parasympathetic innervation of the ventricles is still unclear. There is a common misconception that the sympathetic nerves innervate all over the heart; while the parasympathetic nerves only innervate the superventricular part of the heart, but not the ventricles. Recent evidence indicates that the cholinergic innervation of the left ventricle is functionally very important in some mammalian species. The present article reviews the evidence of vagal control in the ventricles from the anatomy and histochemistry, molecular biology, and function areas. Additionally we overview the vagal (muscarinic) regulation of cardiac contractile function and its signal transduction. PMID- 16344890 TI - Embryonic stem cells cultured in biodegradable scaffold repair infarcted myocardium in mice. AB - Our previous findings demonstrated that directly injecting embryonic stem cells (ESCs) into ischemic region of the heart improved cardiac function in animals with experimental myocardial infarction (MI). Tissue engineering with stem cells may provide tissue creation and repair. This study was designed to investigate the effectiveness of grafting of ESC-seeded biodegradable patch on infarcted heart. MI in mice was induced by ligation of the left coronary artery. Mouse ESCs were seeded on polyglycolic-acid (PGA) material patches. Three days after culture, an ESC-seeded patch was transplanted on the surface of ischemic and peri ischemic myocardium. Eight weeks after MI operation and patch transplantation, hemodynamics and cardiac function were evaluated in four (sham-operated, MI, MI + cell-free patch, and MI + ESC-patch) groups of mice. The blood pressure and left ventricular function were significantly reduced in the MI animals. Compared with MI alone and MI + cell-free patch groups, the animals received MI + ESC-seeded patches significantly improved blood pressure and ventricular function. The survival rate of the MI mice grafted with MI + ESC-seeded patches was markedly higher than that in MI alone or MI + cell-free patch animals. GFP-positive tissue was detected in infarcted area with grafting of ESC-seeded patch, which suggests the survivors of ESCs and possible myocardial regeneration. Our data demonstrate that grafting of ESC-seeded bioabsorbable patch can repair infarcted myocardium and improve cardiac function in MI mice. This novel approach of combining stem cells and biodegradable materials may provide a therapeutic modality for repairing injured heart. PMID- 16344891 TI - [Immunogenicity of recombinant human zona pellucida-3 peptides expressed in E. coli and efficacy of their antisera to inhibit in vitro human sperm-egg binding]. AB - The present study was aimed to analyze the immunogenicity of recombinant human zona pellucida-3 peptides (r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348)) expressed in E. coli through immunizing rabbits, and to evaluate the efficacy of their polyclonal antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding respectively. Male New Zealand rabbits were immunized using r huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) as antigen respectively, which was purified through an improved method of preparative gel polyacryulamide gel electrophoresis. The antibody response level of r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) immunogen in rabbits was determined by ELISA using mouse ZP3-5 (amino acid sequence(137 approximately 150) being completely conserved with huZP3(138 approximately 151) sequence) and specific huZP3-14 (amino acid sequence(327 approximately 340)) synthetic peptides as coating antigens respectively. The immunoreactivity and specificity of the anti-r-huZP3a(22 approximately 176) and anti-r-huZP3b(177 approximately 348) antisera with each r-huZP3 peptides, were tested by immunoblot and immunohistochemistry (using native huZP and human ovary section) respectively. A competitive hemizona assay (HZA) was used to evaluate the efficacy of the antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding. Both r-huZP3 peptides were able to induce higher antibody titers in rabbits. Each antiserum could specifically recognize or bind to each target r-huZP3 peptide expressed in E. coli and native human ZP in vitro. The antisera also inhibited sperm-egg binding in the HZA. These results show that r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) are of strong immunogenicity. They can be used to develop a kit for detecting whether there are autoantibodies to zona pellucida in unexplained infertile women, and their antisera might be useful tools for determining minimal B-cell epitope sequences of several known huZP3 epitope peptides. PMID- 16344892 TI - Ouabain stimulates slowly adapting pulmonary stretch receptors. AB - Ouabain, a Na(+)/K(+)-ATPase inhibitor, induces slowly adapting pulmonary stretch receptors (SARs) to discharge paradoxically. Paradoxical discharge is characterized by increased SAR activity during lung deflation coupled with silence during lung inflation. We hypothesized that over-excitation silences the SARs. Accordingly, if cyclic inflation pressure was reduced so as to lower SAR stimulation, paradoxical discharge would be prevented. In the present study, single-unit activity of SARs was recorded in anesthetized, open-chest and mechanically ventilated rabbits with positive-end-expiratory pressure (PEEP). After microinjection of ouabain into the receptive field, SAR activity initially increased and then gradually became paradoxical. During paradoxical cycling, SAR activity started and stopped abruptly, oscillating between high frequency discharge during lung deflation and silence during peak inflation. Removing PEEP reduced basal cyclic stimulation and returned the discharge pattern to normal, that is, SAR activity was highest at peak inflation pressure but silent during deflation. It is speculated that stretching SARs causes Na(+) influx, producing generator potential (GP). Normally, GP recovers by Na(+) extrusion via Na(+)/K(+) ATPase. Ouabain inhibits the ATPase, which limits Na(+) extrusion, and thus sustains the GP. Therefore, after ouabain microinjection, lung inflation will further increase GP, causing over-excitation to silence the SARs. PMID- 16344893 TI - Tramadol and dihydroetorphine produce synergistic analgesic effect and postpones acute opiate tolerance in rats. AB - The present study investigated whether a co-application of tramadol (TRA) and dihydroetorphine (DHE) would exert a synergy in analgesic effect and delay acute tolerance development. Intraperitoneal injection of TRA (in mg) and subcutaneous injection of DHE (in ng) were delivered in fixed proportions (1:6.25, 1:12.5, 1:25, 1:50, 1:100, and 1:200). The effect of analgesia was accessed by tail-flick test and analyzed with isobolographic analysis. For test of acute tolerance, six successive injections of either TRA (20 mg/kg) alone, DHE (1 000 ng/kg) alone, or a combination of TRA (20 mg/kg) and DHE (250 ng/kg) were administered. We found that (1) except for 1 mg : 6.25 ng and 1 mg : 50 ng, combinations, all the other ratios produced a significant synergy in their analgesic effect; (2) the effect of analgesia induced by repeated TRA plus DHE injections lasted significantly longer, indicating a slower onset of acute tolerance. These results indicate that TRA and DHE injections in certain dose ratios can induce synergistic analgesia, which is resistant against the development of acute tolerance. PMID- 16344894 TI - cDNA microarray analysis of spinal cord injury and regeneration related genes in rat. AB - The acute traumatic spinal cord injury (SCI) is a commonly seen and severe case in clinic. However, the repair and regeneration of injured spinal cord is limited. This is likely due to that different kinds of factors are involved in regeneration after SCI. In the present study, we used complementary DNA microarray consisting of 4 041 specific probes from rat to identify genes that were differentially expressed after SCI. The animals were subjected to complete transection injury of the thoracic spinal cord (T8-T9). Sham operated animals received only a laminectomy. Four and a half days later, rat spinal cord was dissected out for total RNA isolation. The fluorescent (Cy3 and Cy5) labeled probes were prepared and hybridized to the microarray. Genes that showed 2-fold difference in SCI tissue were identified. Sixty-five up-regulated genes consisted of 21 known genes, 30 known expressed sequence tags (ESTs) and 14 unknown genes. Seventy-nine down-regulated genes comprised 20 known genes, 42 known ESTs and 17 unknown genes. In 41 differentially expressed known genes, 5 up-regulated genes, i.e., tissue inhibitor of metalloproteinase 1 (Timp1), transgelin (Tagln), vimentin (Vim), Fc gamma receptor, cathepsin S (Ctss), and 3 down-regulated genes, i.e., stearyl-CoA desaturase, coagulation factor II (F2), endosulfin alpha (Ensa), were further confirmed by reverse transcription polymerase chain reaction (RT-PCR). These genes may play a role in the response to tissue damage or repair following SCI and characterization of them might be helpful to elucidate the molecular mechanisms of spinal cord injury and regeneration. PMID- 16344895 TI - [Telomerase: the expression and regulatory mechanisms in preovulatory ovarian granulosa cells]. AB - The objective of this study was to analyze the expression of telomerase in granulosa cells and the influential factors in telomerase expression. TRAP-ELISA (telomeric repeat amplification protocol-enzyme linked immunoadsordent assay) method was used to study the expression and control of telomerase in rat preovulatory ovary. We also used radioimmunoassay (RIA) to determine the expression of estradiol (E(2)) and progesterone (P(0)). Furthermore we used MTT to study the proliferation of ovarian granulosa cells. Telomerase activity was significantly enhanced by human chorionic gonadotropin (HCG), follicle stimulating hormone (FSH), verapamil and dbcAMP, and was significantly reduced by antisense-c-myb oligodeoxynucleotide (anti-c-myb ODN) treatment. RIA was used to determine the secretion of P(0) and E(2) in all these cell culture media. We found that the secretion of these two hormones was increased when verapamil and FSH were added; no change after adding HCG and dbcAMP; and reduced when anti-c myb was added. In MTT assay, we found that the antisense hTERT ODN significantly inhibited the proliferation of ovarian granulosa cells. These results demonstrate that telomerase activity is present in ovary antral granulosa cells and its activity is controlled by FSH, HCG, verapamil and anti-c-myb, and is directly related with the function of proliferation. PMID- 16344896 TI - [Changes of somatostatin and expression of somatostatin receptor in small intestine and liver tissues during macaque development]. AB - Intestinal tract, which produces more than fifty kinds of gut peptides, is regarded as the largest endocrine organ. With regard to the gut peptides, a number of studies were focused on their structure, function and the roles in some diseases. The changes in output or distribution of gut peptides in the intestinal tract during development have been largely unknown. This study was aimed to investigate the changes of somatostatin (SST) and somatostatin receptor 2 (SSTR2) in small intestinal and hepatic tissues during the development of macaque. The tissue samples of small intestine, liver or blood samples from peripheral and portal vein of 4 macaques in 6-month fetus, 2-day neonate, 45-day neonate and adult were obtained after anesthetization. The concentrations of SST in blood or tissues of macaques were measured by radioimmunoassay. The distributions of SST in small intestinal or hepatic tissues were visualized by immunohistochemical staining. The expression of SSTR2 was detected by in situ hybridization. SST concentration of intestinal tissue in 6-month-old macaque was (27.3+/-16.6) ng /mg protein and light positive staining of SST was localized in mucosal crypts but negative in muscle layer. The intestinal concentration of SST increased gradually with macaque development and reached to the peak [(120.1+/-35.3) ng /mg protein] in adult. It was significantly higher than that in fetus (P<0.01). Strong positive staining of SST was found in both mucosal crypts and myenteric nerve plexus of adult animal. SSTR2 was obviously expressed in intestinal epithelium of fetus but its expression was greatly reduced in epithelium and was shifted to mucosal crypts when grown to adult. Negative staining of SSTR2 in muscle layer of fetal or neonatal macaque turned to be positive in myenteric nerve plexus of adult. The levels of SST or SSTR2 in liver decreased gradually during development. SST concentrations of small intestinal tissue kept significantly higher than those of hepatic tissues in the macaque developing stages. SST levels of portal vein were also maintained significantly higher than those of peripheral blood in the macaque developing stages. In conclusion, the level of SST and expression of SSTR2 in mucosal crypt increased gradually with macaque development. SST from intestinal tract was quickly degraded in portal vein before entering into liver. SST positive myenteric nerve plexus was visualized only in mature macaque. PMID- 16344897 TI - Role of histamine in airway remodeling of asthmatic guinea pig. AB - To investigate the role of histamine in airway remodeling, 50 healthy guinea pigs were divided into 5 groups: control group: nebulized inhalation of distilled water for 8 weeks; asthma model group: nebulized inhalation of ovalbumin (OVA) for 8 weeks after sensitization; continued asthma model group: nebulized inhalation of OVA for 14 weeks after sensitization; histamine group: nebulized inhalation of OVA for 14 weeks after sensitization and histamine was added in the last 6 weeks; antagonist group: nebulized inhalation of OVA for 14 weeks after sensitization and histamine receptor antagonists were added in the last 6 weeks. For each group, the concentration of histamine, sodium ion (Na(+)), chlorine ion (Cl(-)), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), pH, actual bicarbonate (AB), standard bicarbonate (SB) in serum, and thickness of airway mucosa, base membrane and smooth muscle were measured and compared with each other. The results showed that: (1) the concentration of histamine in serum and the thickness of airway increased, the following order was, the control group, the asthma model group, the continued asthma model group and histamine group (P<0.01); and the concentration of histamine in serum and the thickness of airway of antagonist group was lower than that of the continued asthma model group (P<0.05, 0.01). (2) PaO2 of the asthma model group was lower than that of the normal control group (P<0.01); PaO2, pH, AB, SB decreased, the following order was, the asthma model group, the continued asthma model group and the histamine group (P<0.01); and PaO2, pH, AB, SB of the antagonist group was higher than that of the continued asthma model group (P<0.01); but for PaCO2, the order was converse (P<0.01); For the concentration of Na(+) and Cl(-) in serum, there was no difference among these groups. It is concluded that: (1) Histamine is one of the mediators in the airway remodeling of asthma. (2) Histamine receptor antagonists may play a role in preventing and treating airway remodeling. (3) There is a negative correlation between the PaO2, pH and the wall thickness of the airway (P<0.01), while a positive correlation between the PaCO2, anion gap (AG) and the wall thickness of the airway (P<0.01). PMID- 16344898 TI - [Role of bcl-2 transcriptional regulation induced by calmodulin I pressure overload rat hypertrophic hearts]. AB - This study was designed to evaluate the role of bcl-2 transcriptional regulation induced by calmodulin I (CaM I) in pressure overload rat hypertrophic hearts. The model of hypertensive Sprague-Dawley rat was established by abdominal aortic constriction. The hearts were collected four weeks after abdominal aortic constriction. Velocity and isopyknic gradient centrifugation was employed to fractionate rat myocardial nuclei. Western blot analysis revealed a marked increase in phosphorylated cAMP response-element binding protein (pCREB) of cardiac hypertrophy group compared with that in control group (P<0.05), while the protein level of cAMP response-element binding protein (CREB) was constant (P>0.05). Immunohistochemistry results showed a significant increase of CaM I protein in cardiac hypertrophy group relative to the control group (P<0.05). Nuclear run off transcription assay displayed a significant increase in bcl-2 mRNA treated with trifluoperazne compared with non-drug treatment (P<0.05). The results obtained suggest that the transcription of bcl-2 is possibly regulated by CaM I hypertrophic rat hearts, and CREB phosphorylation seems to be a minor factor in bcl-2 transcriptional regulation. PMID- 16344899 TI - Multiple factors contributing to lipopolysaccharide-induced reactivity changes in rabbit pulmonary artery. AB - To explore the underlying mechanism(s) of pulmonary arterial hypertension in endotoxic shock, the roles of N-acetylcysteine (NAC), nitric oxide (NO) and carbon monoxide (CO) were investigated. Pulmonary arterial rings (3-mm width) were prepared from 24 rabbits. Lipopolysaccharide (LPS), after 7-hour incubation, decreased the endothelium-dependent relaxation response of the arterial ring (pre contracted with phenylephrine) to acetylcholine (1 mumol/L), but did not affect the endothelium-independent relaxation response to sodium nitroprusside. The LPS effects were reduced by a concomitant incubation with the free radical scavenger (NAC), NO donor (L-arginine), and CO donor (hemin), respectively. On the other hand, the LPS effects were enhanced by applying heme oxygenase-1 (HO-1) inhibitor (zinc protoporphyrin) to block CO production. The response to acetylcholine changed from relaxation to contraction, however, the contractile response to phenylephrine increased significantly after pre-incubation with nitric oxide synthase (NOS) inhibitor (L-NAME) to block NO production, confirming the importance of CO and NO. These results show that LPS impairs endothelium dependent relaxation of the pulmonary artery, which can be greatly reduced by the antioxidant, or by supplying with NO and CO. Thus, multiple factors are involved in this model of endotoxin-induced pulmonary hypertension. PMID- 16344900 TI - Cardiac hypertrophy induced by prostaglandin F(2alpha) may be mediated by calcineurin signal transduction pathway in rats. AB - In this paper, we studied the relationship between the prostaglandin F(2alpha) (PGF(2alpha))-induced cardiac hypertrophy and calcineurin (CaN) signal transduction pathway in vivo and in vitro. Male Sprague-Dawley rats were given a single i.p. injection with monocrotaline (MCT) (60 mg/kg) and then given orally with celecoxib (20 mg/kg) or vehicle once a day for 14 d before (from d 1 to d 14) or after (from d 15 to d 28) right ventricular hypertrophy (RVH) was formed. Body weight (BW), right ventricular weight (RV), left ventricular with septum weight (LV), as well as lung weight were determined. RVH index (RVHI=RV/LV), RV/BW, and lung weight/BW were calculated and histological changes were observed with transmission electron microscope. PGF(2alpha) level, atrial natriuretic peptide (ANP) and CaN mRNA expressions, expression of CaN and its downstream effectors, NFAT(3) and GATA(4) protein were assayed by EIA kit, RT-PCR, and Western blotting, respectively. The cardiomyocyte hypertrophy in primary culture induced by PGF(2alpha) (0.1 micromol/L) was evaluated by measuring the cell diameter, protein content, and ANP mRNA as well as CaN mRNA expressions. It was found that 14 d or 28 d after MCT was given, the RVHI, RV/BW, and lung weight/BW were significantly increased by 47%, 53% and 118%, and by 64%, 94% and 156%, respectively; at the same time PGF(2alpha) levels in RV tissue were increased by 44% and by 51% with increasing RVHI, and elevated expressions of ANP and CaN mRNA, as well as CaN, NFAT(3) and GATA(4) proteins in a positive correlation manner. Furthermore, some histological injuries were found in RV tissue. Celecoxib, a cyclooxygenase inhibitor, obviously blunted the elevation of RVHI, RV/BW, and lung weight/BW no matter it was given before or after RVH. In vitro experiments showed that 0.1 micromol/L PGF(2alpha) significantly increased the cardiomyocyte diameter and protein content, and promoted ANP and CaN mRNA expressions, which was blocked by cyclosporin A, a CaN inhibitor. Our results indicate that PGF(2alpha) may be involved in cardiac hypertrophy induced by MCT in rats through CaN signal transduction pathway. PMID- 16344901 TI - Gender-related differences of ventricular repolarization in LQT2 rabbit model. AB - To explore the cellular mechanism responsible for the gender-related differences of ventricular tachyarrhythmias in long QT syndromes (LQTS), we observed the characteristics of pre-existing electrophysiological heterogeneity and dynamics of ventricular repolarization in different gender of LQT2 rabbit models. The intracellular floating microelectrodes technique was used to record transmembrane action potentials simultaneously from epicardial and endocardial sites of the arterially perfused rabbit left ventricular wedge preparation; in the mean time, the electrocardiogram (ECG) was also recorded. The wedge preparations were placed in a small tissue bath and arterially perfused with normal Tyrode's solution (control group), 100 mumol/L dl-sotalol Tyrode's solution (LQT2 model group), and 100 mumol/L dl-sotalol plus 3.0 mmol/L KCl Tyrode's solution (LQT2 model plus hypokalemia group), buffered with 95% O2 and 5% CO2 [(36.0+/-1) degrees C]. Double diastolic threshold currents were delivered with basic cycle length (BCL) 2 000, 1 000 and 500 ms (S1), respectively, to record transmembrane action potentials and transmural ECG. Each driving train of S1 is with 20 beats. To determine action potential duration restitution (APDR) curves, the S1-S2 programmed stimuli were used. The tissue was paced at 1 000 ms and 500 ms cycle length (S1) for eight beats, followed by a single premature stimulus (S2). The S1 S2 coupling interval was progressively shortened with 10 ms decrements until the S2 failed to capture. The results showed that female rabbits exhibited significantly longer transmural dispersion of repolarization (TDR) and steeper maximal slopes of APDR curves than that in male rabbits at same pacing rates (P<0.05), and which were pacing rate-dependent. In the condition of dl-sotalol plus hypokalemia, the TDR and the maximal slopes of APDR curves were significantly increased in comparison with that in the control group (P<0.01). At a BCL of 1 000 ms of seven experiments, one female showed torsade de pointes (TdP) in the LQT2 model group; five females and two males showed TdP in LQT2 model plus hypokalemia group, showing significant gender-related differences (P<0.05). The present findings suggest that the pre-existing electrophysiological and dynamic heterogeneity in the LQT2 model shows an obvious gender-related difference and pacing rate-dependence. Both increased TDR and steepness of APDR in female rabbits are possibly the major factors which prompt the TdP generation in female LQT2 rabbits more easily than in male rabbits. PMID- 16344902 TI - [Caspase-3 plays a required role in PC12 cell apoptotic death induced by roscovitine]. AB - Roscovitine is a specific inhibitor of cyclin-dependent kinases (cdks) cdc2/cyclin B, cdk2/cyclin A, cdk2/cyclin E and cdk5/p35. The studies on the enzyme inhibitory properties and cellular effects of roscovitine revealed that it arrests cells in G(2)/M and G(1)/S phase, inhibits the proliferation of mammalian cells and induces cell death. However, the characteristics of cell death and exact mechanism by which this cdk inhibitor kills transformed cells are unknown. We previously investigated that the roscovitine induces apoptotic death of mitotic PC12 cells. The present study was to identify whether the roscovitine induced death is related with the specific elements of caspases in pathway of apoptosis. The morphological data of caspase-3 immunofluorocytochemistry double staining with hoechst 33342 indicated that apoptotic nuclei were identified as nuclei with chromatin condensation and nuclear fragmentation, and that caspase-3 active p17 subunit co-existed in PC12 cells treated with roscovitine 50 micromol/L for 4 h. The number of the caspase-3 positive cells increased significantly to about 42%, as compared with the normal control (P<0.001). The data of MTT assay showed that the number of viable cells treated by roscovitine (50 micromol/L) alone for 12 h was 29.03%, of the untreated controls. Both a broad-spectrum caspase inhibitor Z-VAD-FMK (50 mumol/L) and a specific caspase-3 inhibitor Z-DEVD-FMK (100 micromol/L) increased viable PC12 cells to 45.16%, (Z DEVD-FMK) and 58.06%, (Z-VAD-FMK), respectively, in the presence of roscovitine. Non-erythroid a-spectrin is a cytoskeleted protein that is a substrate of caspase 3 cysteine proteases. To confirm the activity of caspase-3 that produced in roscovitine (50 micromol/L for 12 h)-induced PC12 cell death, activated caspase-3 specific 120 kDa spectrin breakdown products (SBDP) were detected by Western bloting using the mouse anti-non-erythroid a-spectrin monoclonal antibody. The mean relative density of bands corresponding to caspase-3 specific SBDP levels were significantly increased in the cytosolic fractions treated with roscovitine, as compared to the normal control (P<0.001). These results indicate that caspase signals, especially caspase-3 signal are necessary for the progression of proliferating PC12 cell apoptotic death evoked by roscovintine. PMID- 16344903 TI - GABAergic neurons innervating the preganglionic cardiac vagal neurons in the dorsal motor nucleus receive tonic glutamatergic control. AB - The glutamatergic innervations and the GABAergic innervations are respectively the major excitatory and inhibitory inputs of preganglionic cardiac vagal neurons (CVNs). Whether and how these two kinds of innervations interact in the regulation of CVNs is unknown. Using retrograde fluorescent labeling of CVNs and voltage patch-clamp technique, we demonstrated that mixed global application of glutamatergic NMDA and non-NMDA antagonists AP(5) and CNQX, while had no effect on the GABAergic synaptic events of the CVNs in the nucleus ambiguus (NA), significantly decreased the GABAergic synaptic events of the CVNs in the dorsal motor nucleus of the vagus (DMNX). These results suggest that the GABAergic neurons preceding the CVNs in the DMNX receive tonic glutamatergic control, whereas the GABAergic neurons preceding the CVNs in the NA receive little, if any, glutamatergic innervations. This differential central regulation of the CVNs in the DMNX from those in the NA might be a possible mechanism that enables the CVNs in the DMNX play different roles from those in the NA in the parasympathetic control of heart rate and cardiac functions. PMID- 16344904 TI - [Microenvironment effect of APA microcapsule on embryonic stem cell]. AB - We undertook a series of studies to evaluate the role of microenvironment during embryonic stem cell (ESC) proliferation and differentiation. In this paper, cell microencapsulation technology was employed, which allows the free exchange of nutrients, oxygen and biologically active products between the entrapped cell and culture medium. We analyzed the feasibility of mouse ESCs in microcapsules and evaluated the growth, metabolic activity and differentiation of ESCs once enclosed in alginate-Ca(2+) microbead, solid or liquefied core alginate-poly lysine-alginate (APA) microcapsule, respectively. We found that ESCs grew gradually in both types of microcapsules, but the appearance of cells was distinctive for each type of capsule. In the case of unliquefied microcapsules, cells created multiple spherical or lens-shaped aggregates. In contrast, the liquefied alginate core allowed the enclosed ESCs to grow together in a clump at the periphery of the capsule. Combined with cell viability and activity of glucose/lactic acid metabolism, the liquefied core of APA might provide more suitable culture conditions for the ESC growth in comparison with the unliquefied type or alginate-Ca(2+). For better evaluating the nature of ESC growth in APA microcapsules in vitro (that is whether or not encapsulated ESCs maintained undifferentiated state while they kept the ability for proliferation), the expression of the typical markers for undifferentiated, dividing ESCs, such as the stage specific embryonic antigen (SSEA-1) and alkaline phosphatase (AP), was detected by immunochemistry and immunofluorescence staining. The results showed that cell aggregates formed in the microcapsule still expressed the marker proteins at a higher level on day 22 in vitro. The expression of gene Oct-4, a transcription factor necessary for maintaining ESCs in an undifferentiated state, was also detected when RT-PCR assay was employed (on day 22 in vitro). In addition, cell aggregates were released from the microcapsules by mechanical disruption and induced into insulin-producing cells. These findings further indicate that most of the ESCs in APA microcapsule maintain their multi-potential even though the culture time prolonged as long as 22 d in vitro. Taken together, APA microcapsule provides a suitable microenvironment that promotes ESCs to maintain their stemness. Therefore, the microenvironment plays an important role in the process of ESC proliferation and differentiation. PMID- 16344905 TI - [Man and his fellow-creatures under ethical aspects]. AB - It is for reasons of age I will have to terminate my work at the Literary Review in the form developed since 1995. The report is being reduced to a concentration of ethically relevant reviews as exemplified in the fourth-quarter issue of ALTEX. This is to ascertain that essential developments in this field will not be overlooked. Insofar, the Literary Review will be continued under the heading "New literature concerning topics of animal ethics". The more central topics of animal ethics are being "used up" the more new questions are being formulated. Thus it was that during the last few years the plant-world, long neglected, was rediscovered and received attention through the publication of important works. Another recent discovery concerns itself with "cognitive ethology" which developed out of the critique of behaviourism and which is dealt with in a separate chapter in this issue. But there is also a "classic" of ethics who has been reviewed and interpreted anew repeatedly. In her book "Albert Schweitzer, a prophet of medical ethics", Heike Baranzke describes Schweitzer's ethics as not sentimental or nostalgic but rather as a radically modern stance, committed to the enlightenment. Manuel Schneider, also, conveys a comprehensive view of Albert Schweitzer's ethics in "Life in the middle of life - the relevance of the ethics of Albert Schweitzer", a book edited by Altner, Frambach, Gottwald and himself in 2005. For this, in particular, he derives a possibility of a physiocentric ethics. By contrast, Beate Weinzierl approaches Schweitzer on a complete personal and human level in "Yearning for nature - access to inner and outer nature with Albert Schweitzer". Wolfgang Senz is undertaking a critical appreciation of Albert Schweitzer's concept of "life" and this, foremost, in the light of Schweitzer's rejection of the Cartesian "I am". In the end, Jean Claude Wolf cannot manage without citing Schweitzer either, referring to him in his not accepting the (western) world's excessive meat consumption as a "cruel necessity". Klaus Peter Joern also enters into Schweitzer's fundamental argumentation in his "A good bye to the disparagement of our fellow creatures". Quotation: "It just cannot go on that Christianity lets the validity of the commandment of love end at the behaviour towards humans, simply because that is the tradition. We rather have to, in this instance, deny the authority of the Bible it's due respect, as this has deemed Schweitzer already inevitable." The killing of animals, irrespective of the motives, remains a critical issue in the understanding (or the lack thereof) with regard to the dignity of all creation. Manuela Linnemann, Charles Patterson, Evelyn Ofensberger and the German Veterinary Association for the Protection of Animals have dealt with this issue extensively. As always, more space is taken up by publications dealing with legal questions and developments. In particular, the conference report of the Protestant Academy at Bad Boll should be mentioned "Animal protection on good constitution". Contributions to this report, edited by the academy, include Hans Georg Kluge's "The governmental aim 'animal protection' and it's implementation in jurisdiction" and Johannes Caspar's "effects of the governmental goal 'animal protection' in the protected area of unconditional fundamental rights". Regarding animal experiments, the last two position papers commissioned by the Foundation for animal-free research (FFVFF), dealing with perspectives of 3R research, have been published within the period of time covered by this report. Alternatives in basic research (with once again rising numbers of experimental animals) and in biomedical education are the themes dealt with by Franz P. Gruber and his co authors Thomas Hartung and David Dewhurst. PMID- 16344906 TI - [The three Rs: looking back ... and forward]. AB - Personal experiences at FRAME and ECVAM are recalled, alongside the evolutionary progress of the Three Rs (replacement, reduction, refinement,) approach to animal experimentation, described in great decades analogous to the Great Ages of Western Civilisation. PMID- 16344907 TI - [The use of transgenic animals in biomedical research in Germany. Part 1: Status Report 2001-2003]. AB - While the German Federal Government has set itself the goal to make an active contribution to reducing animal experiments, the use of transgenic animals in biomedical research continuously increases every year. It is against this background that the study at hand aimed at providing an overview over the goals and the contents of research projects performed in Germany, in the course of which transgenic animals were produced or used in experimental procedures. Specifically, it was envisaged to spell out those specific areas of research, for which transgenic animals mainly were being used. Subsequently it was evaluated whether the research goals revealed might also be pursued with non animal test methods. In a literature survey, a total of 577 scientific publications relevant for the purposes of the study were collected. This material enables conclusions on those scientific areas, in which transgenic animals are used, applying to fundamental research, but not on their use in routine procedures in applied research or for the maintenance of transgenic breeds, since such purposes do not tend to be the subject of publications in scientific journals. According to the topics covered by the publications, main areas of biomedical research with transgenic animals can be found in the fields of neurobiology, immunology, cardiology, embryology and oncology. However their use can be discerned in all other areas of fundamental biomedical research as well. In accordance with the official German laboratory animal statistics, the vast majority of transgenic animals used were mice, followed by rats and pigs. Additionally, singular research projects with fish, rabbits and chicken were recorded. (In the official German laboratory animals statistics, very small numbers of transgenic hamsters, sheep and amphibians were also recorded in the past years.) A high percentage of the rats were used in cardiovascular research, whereas transgenic pigs as a rule were produced and bred as organ donors in xenotransplantation research. The majority of research projects either dealt with the experimental use of already established transgenic animal lines, or they described that transgenic animals specifically were produced for the purpose of the respective research project. Mostly, transgenesis was initiated by inserting the foreign gene into the germ cell genome. In some research projects, it was reported that the transgenic material was inserted into normally bred animals some time after parturition. PMID- 16344908 TI - Reactivation of creatine kinase by dithiothreitol prior to use in an in vitro translation extract. AB - BACKGROUND: In vitro protein synthesis on exogenous messenger ribonucleic acids can be performed in various systems including cytoplasmic extract from eukaryotic cells, rabbit reticulocyte lysate and wheat germ extract. For optimal translation, an energy regeneration system based on creatine phosphate and creatine kinase is commonly employed for the regeneration of the endogenous adenosine triphosphate pools. Creatine kinase purchased from various commercial suppliers can be partially oxidised. Oxidised creatine kinase is not biologically active and might not allow the efficient initiation of translation of exogenous mRNAs in eukaryotic cell extracts in vitro. RESULTS: We successfully used dithiothreitol to reduce and therefore reactivate commercially available creatine kinase. When employed in cytoplasmic extracts obtained from eukaryotic cells grown in monolayers, the reactivated creatine kinase restored translation of the exogenous mRNAs. CONCLUSION: Lyophilised creatine kinase obtained from commercial suppliers can be purchased as an oxidised monomer. The reactivation of creatine kinase using a reducing agent such as dithiothreitol restores the biological activity of this enzyme. This procedure might therefore be extended to various other in vitro conditions and biological systems in which the maintenance of an efficient ATP-regenerating system is critical. PMID- 16344909 TI - [Ethics as a scientific basis for animal protection - on the advancement and amendment of the Codex Veterinarius]. AB - Seven years ago, the veterinary association for animal protection (Tieraerztliche Vereinigung fuer Tierschutz e.V., TVT) in Germany first published an orientation guide on the ethics of animal protection aimed at the entire veterinary profession: the "Codex Veterinarius - ethical guiding principles on veterinary action for the good and the protection of animals". The dilemmas of the veterinary profession have not changed since then, but as the principle that "nothing can be so good that it cannot be improved" also applies to the Codex, the two authors have attempted to optimise the few weak spots of the Codex. Joerg Luy sees the task of the Codex in surpassing the animal protection law in two respects: on the one hand regarding moral aspects that cannot be regulated (e.g. respect for the "intrinsic value" of the animal, which is greater than its "utilitarian value") and on the other hand in cases where the law does not live up to its own goals (e.g. regarding the inconsistent regulation on causing "longer term or repeated grave pain or suffering"). PMID- 16344910 TI - Ukraine and Russia: major InterNICHE outreach. AB - During October 2005, InterNICHE National Contacts Anya Yushchenko and Lena Maroueva, and Co-ordinator Nick Jukes, began a large-scale promotion of alternatives across the Ukraine and Russia. The InterNICHE outreach produced a number of positive results: presentations, demonstrations and training in alternatives to over 500 teachers and students; agreements at two institutes to replace harmful animal use in education across a whole department and faculty respectively; information gathering, and discussions with teachers about potential future replacement; reaching a massive audience through positive media coverage; and empowerment of campaigners, including InterNICHE National Contacts who successfully organised the majority of the activities. To continue the successes, funds to enlarge the Russian micro-Loan System of alternatives and to make donations of alternatives to institutes are urgently needed. Support for broadening the InterNICHE impact across Ukraine, Russia and Asian republics through the distribution of translated literature, video, freeware alternatives and web-based resources is also required. Such activity not only supports the development of a progressive, humane education, but impacts positively on animal use in research and testing by creating an environment more conducive to alternatives in general. This is important now as animal testing laboratories consider relocating in or sub-contracting to ex-Soviet countries and in central Asia. New InterNICHE connections have already been made with Belarus, Armenia, Azerbaijan and Kyrgyzstan, and there are many open doors. PMID- 16344911 TI - Complex formation of the laminin-5 gamma2 chain and large unspliced tenascin-C in oral squamous cell carcinoma in vitro and in situ: implications for sequential modulation of extracellular matrix in the invasive tumor front. AB - Invasion and metastasis in oral squamous cell carcinoma (OSCC) are associated with changes in the extracellular matrix (ECM). We have previously shown an extracellular co-deposition of laminin-5 (Ln-5) and large unspliced tenascin-C (Tn-C(L)) in OSCC. Using a co-culture model of hTERT-BJ1 fibroblasts and the OSCC cell line PE/CA-PJ15, we demonstrate in the present study that Ln-5 and Tn-C(L) are not only co-deposited, but also form a physical complex which can be recovered by co-immunoprecipitation. In agreement with these results, examination of OSCC tissue specimens of different malignancy grade by means of confocal laser scanning microscopy revealed different patterns of Ln-5 and Tn-C(L) co localization implicating complex formation also in vivo. A ribbon like co localization was detected in subepithelial basement membranes around well differentiated OSCC parts and tumor clusters. Furthermore, a fibrillar Ln-5 gamma2 chain/Tn-C(L) co-localization occurred in the carcinoma stroma beneath tumor clusters. Additionally, at the site of ruptured basement membranes there were dot or strand like co-deposits of both molecules, but co-localizations were only rarely detectable. These different patterns may reflect a sequential modulation and reorganization of the ECM in the tumor/stroma interface as it occurs in different stages of OSCC invasion. PMID- 16344912 TI - Characterization of the psychrotolerant acetogen strain SyrA5 and the emended description of the species Acetobacterium carbinolicum. AB - A psychrotolerant, obligate anaerobic, acetogenic bacterium designated strain SyrA5 was isolated from black anoxic sediment of a brackish fjord. Cells were Gram-positive, non-sporeforming rods. The isolate utilized H(2)/CO(2), CO, fructose, glucose, ethanol, ethylene glycol, glycerol, pyruvate, lactate, betaine and the methyl-groups of several methoxylated benzoic derivatives such as syringate, trimethoxybenzoate and vallinate. The optimum temperature for growth was 29 degrees C, whilst slow growth occurred at 2 degrees C. The strain grew optimally with NaCl concentrations below 2.7% (w/v), but growth occurred up to 4.3% (w/v) NaCl. Growth was observed in the range from pH 5.9 to 8.5, optimum at pH 8. The G+C content was 44.1 mol%. Based upon 16S rRNA gene sequence analysis and DNA-DNA reassociation studies, the organism was classified in the genus Acetobacterium. Strain SyrA5 shared a 16S rRNA sequence similarity with A. carbinolicum of 100%, a fthfs gene (which codes for the N5,N10 tetrahydrofolate synthetase) sequence identity of 98.5-98.7% (amino acid sequence similarities were 99.4-100%) and a RNA-DNA hybridization homology of 64-68%. Despite a number of phenotypic differences between strain SyrA5 and A. carbinolicum we propose including strain SyrA5 as a subspecies of A. carbinolicum for which we propose the name Acetobacterium carbinolicum subspecies kysingense. The type strain is SyrA5 (=DSM 16427(T), ATCC BAA-990). PMID- 16344913 TI - Sexual health and the gender gap. PMID- 16344914 TI - Foot deformities in rheumatoid arthritis and relevance of foot function index. AB - The aim of this study were to assess what type of foot deformities are found in rheumatoid arthritis (RA) patients, to detect frequency of deformities, and to evaluate deformities affecting Foot Function Index (FFI) and patient functional capacity. Anteroposterior and lateral weight-bearing radiographs of 156 feet of 78 patients who had RA for > or =2 years and of 76 feet of 38 healthy controls were studied. We measured hallux valgus angle, intermetatarsal angle between first and second (M1/2) and intermetatarsal angle between first and fifth (M1/5) on anteroposterior radiographs, and calcaneal pitch on the lateral radiographs. We examined the feet of all RA patients and healthy controls for hallux rigidus, cock-up deformity, clawing toe, and mallet finger, and measured calcaneal valgus angle. FFI, comprised of pain, disability, and activity limitation subscales, was administered to all RA patients. Their Steinbrocker Functional Class (SFC) and Health Assessment Questionnaire (HAQ) scores were determined. We determined frequency of deformities as 96.2% in RA patients and 97.4% in controls by radiological and physical examination (p>0.05). The frequency of each deformity was markedly increased in RA patients, with the exception of calcaneal valgus deformity. There was significant correlation between SFC and HAQ with FFI and subscales (respectively, r=0.46, p=0.001; r=0.67, p=0.001). For FFI and subscales, HAQ was the most important predictor factor, followed by gender and hallux rigidus. Foot deformities are seen very frequently in RA. These deformities may affect patient functional foot, especially hallux rigidus and calcaneal valgus. PMID- 16344915 TI - Phase II study of dose-dense sequential doxorubicin and docetaxel for patients with advanced operable and inoperable breast cancer. AB - Rapid sequential delivery of doxorubicin 75 mg/m2 q 2 weeksx3 cycles followed by docetaxel 100 mg/m2 q 2 weeksx3 cycles, with filgrastim support was evaluated in patients with inoperable and large operable breast cancers who were not initially candidates for breast conservation therapy. Postoperative CMF chemotherapy and/or radiation were administered based on surgical findings. Median age of the 39 enrolled patients was 47 (range 27-59), stage IIA (6 patients), IIB (14 patients), IIIA (10 patients), IIIB (9 patients), and 23 patients (59%) had clinical nodal involvement. The average bidimensional tumor size before treatment was 30 cm2. Clinical responses included 13 (33%) complete responses, 23 (59%) partial responses, 1 stable disease, and 2 progressive disease, for an overall response rate of 92%. Clinical response rate was 11/13(85%) in HER2/neu positive patients compared to 25/26 (96%) in tumors that did not express HER2/neu. Twenty patients (51%) underwent breast conservation surgery. Pathologic tumor response at the time of definitive surgery included 4 pathologic CR (pCR, 10%), 4 microscopic invasion (pINV), and 14 (36%) pathologically negative axillary nodes. pCR was not observed in any HER2/neu positive patients. 5/39 patients were unable to complete all cycles of docetaxel and 8 patients required dose reduction of docetaxel due to development of grade 3-4 mucositis and hand-foot syndrome. This observation prompted a protocol change requiring 3 weeks between doxorubicin and docetaxel. Primary chemotherapy with dose-dense doxorubicin and docetaxel given sequentially is well tolerated and allows a high rate of breast sparing in patients with large breast cancers. PMID- 16344916 TI - The role of CXCR4 receptor expression in breast cancer: a large tissue microarray study. AB - The chemokine receptor CXCR4 is an important factor in the migration, invasiveness, metastasis and proliferation of breast cancer cells. We have retrospectively analyzed the levels of expression of CXCR4 in a large cohort of breast cancers and pre-invasive breast samples linked to clinical data. A total of 1808 invasive breast carcinomas and 214 pre-invasive breast samples could be analyzed in correlation with basic clinico-pathological data such as hormone receptor status, HER2 status and tumor grade. The majority of breast cancers expressed either nuclear or cytoplasmic staining or both. CXCR4 cytoplasmic expression was associated with parameters of tumor aggressivity (tumor grade and lymph node status) and had prognostic value (age-adjusted hazard ratio=1.73; Confidence Interval: 1.07-2.77) with respect to disease-specific survival. CXCR4 positivity in the cytoplasm but not the nucleus was associated with HER2 expression and amplification as well as with hormone receptor negativity (both ER and PR). The percentage of nuclear staining increased from normal breast tissue (20%) to ductal carcinoma-in-situ DCIS (43%) to invasive cancer (67%) while CXCR4 was expressed in the cytoplasm of 67% of (DCIS) cases (double that in normal breast samples), suggesting an important role in breast tumor progression. The CXCR4 receptor is expressed in many breast cancers, justifying its development as a therapeutic target in breast cancer patients. Its cytoplasmic expression is associated with breast tumor progression, suggesting potential value as a diagnostic marker. PMID- 16344917 TI - Propagation of population PK and PD information using a Bayesian approach: dealing with non-exchangeability. AB - The aim of this work is to implement a conservative prior that safeguards against population non-exchangeability of prior and data likelihood, in the framework of population pharmacokinetic/pharmacodynamic analysis, incorporating multi-level hierarchical modelling. Three different exercises were performed: (i) we investigated the use of parametric priors in the multilevel hierarchical modelling framework; (ii) we assessed the average performance of the multilevel hierarchical model compared to the standard mixed effect model, considering also some interesting extreme cases and (iii) we implemented an application with a small Proof of Principle (PoP) study, which demonstrates the propagation of information across PD studies using multilevel modelling. Fitting with the 4 level model and informative parametric priors performed similar to a meta analysis of the test datasets combined with the datasets that the priors came from, demonstrating that parametric priors can be used alternatively to meta analysis. Further, the 4-level model gave posterior distributions which had larger uncertainty but at the same time were unbiased, compared to the 3-level model, and therefore implements a more conservative prior in a formal way, which is appropriate when the prior and the test populations are not exchangeable. For the application with the PoP study, the statistical power of detecting the difference in potency between two drugs, when inter-study variability was present, was greater when an extra level in the hierarchical model to account for it, was used. In conclusion, by applying the prior one hierarchical level above the level of the parameters of interest, we implemented a more conservative prior, compared to applying the prior directly on the parameters of interest. The approach is equivalent to Bayesian individualization, offers a safeguard against bias from the prior and also avoids the danger of the data being overwhelmed by a strong prior. PMID- 16344919 TI - Effects of the benzoxazolinone BOA, selected degradation products and structure related pesticides on soil organisms. AB - The benzoxazolinone BOA and the degradation products APO, AAPO and HPAA, as well as four structure related compounds to BOA, were tested for their lethal and sublethal effects on the collembola Folsomia candida and on the carabid beetle Poecilus cupreus applying validated standard laboratory methods. According to the results of the recommended risk assessment for plant protection products (EPPO, 2003), BOA and its structure related compounds carbendazim, benoxacor and benazolin (and additionally the formulated product Cresopur) were classified as low risk compounds for both test organisms. Phosalone was of high risk for F. candida. APO, AAPO and HPAA were of medium risk for F. candida, but were rated as low risk compounds for P. cupreus at the tested rates of about 2 mg/kg substrate. However, as the BOA degradation products have been found to occur at field rates below 0.2 mg/kg substrate or are often even not detectable, it is assumed that their potential risk for both non-target soil organisms in the field will be acceptable. PMID- 16344918 TI - Systemic high-dose intravenous methotrexate for central nervous system metastases. AB - BACKGROUND: Treatment options for patients with recurrent central nervous system (CNS) metastases are limited. Rapid infusion of high-dose intravenous methotrexate (HD IV MTX) penetrates the blood-brain barrier (BBB) and has reported activity in leptomeningeal metastases. METHODS: Medical records were reviewed for all patients treated with HD IV MTX (3.5 g/m2) for CNS parenchymal or leptomeningeal metastases. Radiographic response rate, survival, and toxicity were determined. RESULTS: Thirty-one women and one man with a median age of 52 years (range 33-76) were treated with a total of 141 cycles (median 4, range 1 13). Twenty-nine patients had breast cancer, and one each had cancer of unknown primary (CUP), squamous cell carcinoma of the head and neck, and non-small cell lung cancer (NSCLC). An objective radiographic response and stable disease were each observed in nine patients (28%), and 13 (44%) patients progressed. Prior treatment with low-dose MTX for systemic disease did not affect response (P = 0.8). The median overall survival (n = 32) was 19.9 weeks (range 2.9-135.4+) with one patient alive at 135.4 weeks. Myelosuppression and elevated serum hepatic transaminases were the most common acute toxicities (21% and 9% of HD IV MTX cycles, respectively). CONCLUSIONS: HD IV MTX is effective in the treatment of CNS metastases with disease control (response or stable) as a best response in 56% of assessable patients. Further study is warranted. PMID- 16344920 TI - Critical issues and further questions about respondent-driven sampling: comment on Ramirez-Valles, et al. (2005). AB - The application of respondent-driven sampling (RDS) is a promising new approach to understanding hidden populations, including those at high risk for HIV infection. The method has significant advantages over other sampling methods, including the possibility that representativeness samples can be accrued. However, the requirements for a respondent-driven sample to achieve representative and to demonstrate its superiority over other methods are quite strict. This report focuses on whether the accompanying paper by Ramirez-Valles et al. fulfills these requirements in terms of the theorems underlying RDS regarding sample recruitment rates, referral patterns, and network sizes. Further, it investigates the assertion that the accrued sample has proven that RDS is superior to more traditional time-location or venue-based sampling methods. Unconvinced that either is the case, the author suggests a method to test if RDS is indeed the gold-standard recruitment strategy for obtaining inclusive and representative samples of hidden populations. PMID- 16344921 TI - Translational medicine: can it really facilitate the transition of research "from bench to bedside"? AB - Translational medicine is intended to facilitate the transition of basic science results to clinical practice, thereby sharing major aspects of clinical pharmacology. Biomarkers need to be developed to achieve this, and their predictive values need to be assessed. Despite all the attempts to increase output from costly pharmaceutical research investments, all stakeholders complain of the decreasing efficiency of drug development processes, and expensive late attritions seem to be seen at increasing rates. How can translational medicine improve this apparent mismatch between effort and tangible result for daily medical practice? What is missing, and where do we stand? PMID- 16344923 TI - Use of antimicrobials and risk of type 1 diabetes in a population-based mother child cohort. AB - AIMS/HYPOTHESIS: The aim of this study was to investigate whether the use of antimicrobials is associated with the risk of childhood type 1 diabetes. MATERIALS AND METHODS: The study population included all children born in Finland between 1996 and 2000 who were diagnosed with type 1 diabetes by the end of 2002. For each case (n=437), four matched controls were selected. Data on diabetes and the maternal use of antimicrobials was derived from nationwide registries. RESULTS: Maternal use of phenoxymethyl penicillins (odds ratio [OR]=1.70, 95% CI 1.08-2.68, p=0.022) or quinolone antimicrobials (OR=2.43, 95% CI 1.16-5.10, p=0.019) before pregnancy was associated with an increased risk of type 1 diabetes in the child, whereas the use of other specific antimicrobials was not related to the risk. The risk was also higher among mother-child pairs where macrolides were used both by the mother before pregnancy and by the child, compared with pairs where neither used macrolides (OR=1.76, 95% CI 1.05-2.94, p=0.032). Maternal use of antimicrobials during pregnancy was not associated with an increased risk. The high use of antimicrobials by the child (more than seven vs seven or less purchases) was related to greater risk (OR=1.66, 95% CI 1.24 2.24, p=0.001). CONCLUSIONS/INTERPRETATION: Overall, the use of antimicrobials before pregnancy, during pregnancy or during childhood was not related to the risk of childhood type 1 diabetes. However, the use of some specific antimicrobials by the mother before pregnancy and by the child may be associated with an increased risk. Further studies are needed to confirm these associations and to elucidate the underlying mechanisms of action. PMID- 16344922 TI - Drug-resistant pneumococcal pneumonia: clinical relevance and approach to management. AB - Community-acquired pneumonia is the most common infectious disease that causes death, with Streptococcus pneumoniae remaining the leading causative pathogen. The worldwide incidence of infections caused by pneumococci resistant to penicillin, macrolides, and other antimicrobial agents has increased at an alarming rate during the past 2 decades. Yet, these agents are still used as first-line empirical therapy in the outpatient setting. There are several reasons for this, including the infrequency of making a pathogen-specific diagnosis, the failure of studies to demonstrate the relevance of resistance, and the infrequency with which clinicians recognize clinical failures. Despite this, there is mounting evidence that supports the practice of using high doses of some antimicrobial agents, a more active antimicrobial agent within a class, or switching to another class of antimicrobial agents when a patient is identified as being at an increased risk of infection with a resistant pneumococcus. There is now information that will allow the physician to identify not only the patient at risk for infection with a resistant pneumococcus but also the antimicrobial class and, in some cases, the agent within the class to which the organism is more likely to be resistant. This will allow clinicians to better define optimal therapy for patients with community-acquired pneumonia. PMID- 16344924 TI - Rosiglitazone-associated pseudotumour cerebri. PMID- 16344925 TI - Insulin control of placental gene expression shifts from mother to foetus over the course of pregnancy. AB - AIMS/HYPOTHESIS: The human placenta is a complex organ situated at the interface between mother and foetus that separates maternal from foetal blood. The placental surfaces exposed to the two bloodstreams are different, i.e. trophoblasts and endothelial cells are in contact with the maternal and foetal circulation, respectively. Both cell types produce high insulin receptor levels. The aim of the present study was to test the hypothesis that spatio-temporal changes in insulin receptor expression in trophoblasts from first trimester to the endothelium at term shift the control of insulin-dependent processes from mother to foetus. METHODS: Global microarray analysis of primary trophoblasts from first trimester and term human placentas and endothelial cells from term human placentas cultured under hyperinsulinaemic and control conditions identified different sets of regulated genes in trophoblasts and endothelial cells. RESULTS: Insulin effects on placental gene expression underwent developmental changes from trophoblasts in the first trimester to endothelial cells at term that were paralleled by changes in levels of activated insulin receptors. The changes in gene regulation were both quantitative (i.e. magnitude of effect) and qualitative (i.e. specific genes affected and direction of regulation). CONCLUSIONS/INTERPRETATION: This spatio-temporal shift in insulin sensitivity throughout pregnancy allows maternal and foetal insulin to regulate different processes within the placenta at different gestational stages, facilitated by compartmentalisation of the insulin response. Thus, by altering the levels and function of insulin receptors in space and time, control of insulin-dependent processes in the human placenta will change from mother to foetus throughout gestation. This will be of particular interest in conditions associated with altered maternal or foetal insulin levels, i.e. diabetes mellitus or intrauterine growth restriction. PMID- 16344926 TI - To: Mathieu C, Gysemans C, Giulietti A, Bouillon R (2005) Vitamin D and diabetes. Diabetologia 48:1247-1257. PMID- 16344928 TI - Nanotechnology: dry versus wet engineering? PMID- 16344927 TI - Luminescent quantum dots in immunoassays. PMID- 16344930 TI - Adaptive control of gait stability in reducing slip-related backward loss of balance. AB - The properties of adaptation within the locomotor and balance control systems directed towards improving one's recovery strategy for fall prevention are not well understood. The purpose of this study was to examine adaptive control of gait stability to repeated slip exposure leading to a reduction in backward loss of balance (and hence in protective stepping). Fourteen young subjects experienced a block of slips during walking. Pre- and post-slip onset stability for all slip trials was obtained as the shortest distance at touchdown (slipping limb) and lift-off (contralateral limb), respectively, between the measured center of mass (COM) state, that is, position and velocity relative to base of support (BOS) and the mathematically predicted threshold for backward loss of balance. An improvement in pre- and post-slip onset stability correlated with a decrease in the incidence of balance loss from 100% (first slip) to 0% (fifth slip). While improvements in pre-slip stability were affected by a proactive anterior shift in COM position, the significantly greater post-slip onset improvements resulted from reductions in BOS perturbation intensity. Such reactive changes in BOS perturbation intensity resulted from a reduction in the demand on post-slip onset braking impulse, which was nonetheless influenced by the proactive adjustments in posture and gait pattern (e.g., the COM position, step length, flat foot landing and increased knee flexion) prior to slip onset. These findings were indicative of the maturing process of the adaptive control. This was characterized by a shift from a reliance on feedback control for postural correction to being influenced by feedforward control, which improved pre-slip stability and altered perturbation intensity, leading to skateover or walkover (>0.05 m or <0.05 m displacement, respectively) adaptive strategies. Finally, the stability at contralateral limb lift-off was highly predictive of balance loss occurrence and its subsequent rapid reduction, supporting the notion of the internal representations of stability limits that could be modified and updated, as a key component in the adaptive control. PMID- 16344929 TI - Persistent monoarthritis of the temporomandibular joint region enhances nocifensive behavior and lumbar spinal Fos expression after noxious stimulation to the hindpaw in rats. AB - Effects of persistent temporomandibular joint (TMJ) inflammation on nociceptive responses of remote bodily areas of the rat were investigated. Monoarthritis of the TMJ region was evoked by the injection of complete Freund's adjuvant (CFA) into the left TMJ region. Rats without injection of CFA into the TMJ region served as controls (non-CFA group). Time spent on licking behavior evoked by the injection of formalin into the left hindpaw and withdrawal thresholds of mechanical stimulation to both sides of the hindpaw were measured during TMJ inflammation for 3 weeks. Furthermore, expression of Fos protein in the lumbar dorsal horn was immunohistochemically investigated following the injection of formalin into the hindpaw during TMJ inflammation. Formalin-evoked nocifensive behavioral activities were significantly enhanced at 10 and 14 days after CFA injection in the late phase, while the withdrawal threshold to mechanical stimulation was significantly decreased bilaterally at 8, 10 and 14 days after CFA injection. Both formalin-evoked licking behavior and mechanical withdrawal thresholds to bilateral hindpaw at 21 days after CFA injection were similar to those in the non-CFA group. The number of Fos-positive neurons in the lumbar dorsal horn ipsilateral to the formalin injection at 1 and 7 days after CFA injection into the TMJ were similar to those in the non-CFA group; however, those were significantly increased in the laminae I-II and V-VI of the lumbar dorsal horn at 14 days after CFA injection. TMJ inflammation for 7 and 14 days alone produced a small number of Fos-expressing neurons in the lumbar dorsal horn. These results provide evidence that persistent unilateral inflammation of the TMJ region causes an increase in behavioral hyperalgesia of the hindpaw, which is attributed to the modulation of neural activities, in part, in the lumbar dorsal horn, likely mediated by supraspinal neural mechanisms. PMID- 16344931 TI - A serum-free medium for colony growth and hyaluronic acid production by Streptococcus zooepidemicus NJUST01. AB - A hyaluronic acid (HA)-producing strain, Streptococcus zooepidemicus NJUST01, can grow in a serum-free agar medium, with starch as exclusive carbon source, but not glucose, sucrose, dextrine, xylose, or lactose. In this starch medium, the strain NJUST01 reproduced successively at 37 degrees C for 60 generations, with no obvious variation on morphology and physiology, but colonies of the strain after 60th generation could not produce a clear hemolytic zone on sheep blood agar plates. Hyaluronic acid production by the strain NJUST01 was analyzed relative to the starch medium. Employing a multifactor cross experiment, an optimum medium revealed for hyaluronic acid production was composed of 5% starch, 0.3% glucose, 0.5% peptone, 0.15% MgSO4, and 2.0% K2HPO4. The amount of HA 6.7 g/l was obtained in batch fermentation on a rotary shaker at 37 degrees C, 220 rpm for 36 h. PMID- 16344932 TI - Rhamnolipid production by a novel thermophilic hydrocarbon-degrading Pseudomonas aeruginosa AP02-1. AB - Thermophilic bacterial cultures were isolated from a hot spring environment on hydrocarbon containing mineral salts media. One strain identified as Pseudomonas aeruginosa AP02-1 was tested for the ability to utilize a range of hydrocarbons both n-alkanes and polycyclic aromatic hydrocarbons as sole carbon source. Strain AP02-1 had an optimum growth temperature of 45 degrees C and degraded 99% of crude oil 1% (v/v) and diesel oil 2% (v/v) when added to a basal mineral medium within 7 days of incubation. Surface activity measurements indicated that biosurfactants, mainly glycolipid in nature, were produced during the microbial growth on hydrocarbons as well as on both water-soluble and insoluble substrates. Mass spectrometry analysis showed different types of rhamnolipid production depending on the carbon substrate and culture conditions. Grown on glycerol, P. aeruginosa AP02-1 produced a mixture of ten rhamnolipid homologues, of which Rha Rha-C10-C10 and Rha-C10-C10 were predominant. Rhamnolipid-containing culture broths reduced the surface tension to approximately 28 mN and gave stable emulsions with a number of hydrocarbons and remained effective after sterilization. Microscopic observations of the emulsions suggested that hydrophobic cells acted as emulsion-stabilizing agents. PMID- 16344933 TI - Lymphocytic alveolitis: A surprising index of poor prognosis in patients with primary Sjogren's syndrome. AB - Twelve years ago we reported that lymphocytic alveolitis [or bronchoalveolar lavage (BAL) lymphocytosis] correlates with clinical pulmonary involvement in primary Sjogren's syndrome (pSS). Our thesis was based on subtle clinical and functional evidence of interstitial lung disease (ILD) in pSS patients with "high lymphocytic alveolitis" (>15% lymphocytes in BAL). This report is a follow-up study of these patients. Basic clinical and functional re-evaluation of the 22 patients with pSS, studied in 1991, emphasized the differences between those with alveolitis and those without alveolitis. There was no significant functional decline. There were, however, two statistically significant differences between the two groups: (1) only patients with BAL lymphocytosis had to be treated with steroids (5/12 vs. 0/10, P < 0.05) and (2) only patients with BAL lymphocytosis had died in the mean time (6/12 vs. 0/10, P < 0.01). The causes of death were various. On only two occasions were they related to respiratory infections while there were no deaths from respiratory failure secondary to ILD. BAL lymphocytosis appears to be a surprisingly serious index of dismal prognosis in patients with pSS. We offer no unifying pathophysiologic mechanism for it and, therefore, all we propose is that BAL is performed early, in as many patients with pSS as possible. These patients should then be followed up systematically, in order to evaluate if BAL lymphocytosis has any pathophysiologic importance in the development of clinically serious pSS, which is serious enough to lead to death. PMID- 16344934 TI - Clinically significant gastro-oesophageal reflux following oesophageal flap repair for oesophageal atresia and tracheo-oesophageal fistula. AB - Clinically significant gastro-oesophageal reflux (GOR) following oesophageal atresia and tracheo-oesophageal fistula (OA/TOF) repair is commonly considered normal sequela after repair. A retrospective review of patients operated on by two consultants was undertaken. All patients underwent oesophageal tailoring and augmentation for reconstruction of their oesophagus. The presence of clinically significant GOR was confirmed by contrast swallows, 24 hour pH study and endoscopy. Clinically significant GOR occurred in 7 (13%) of the 54 patients operated for OA and TOF. Two patients responded to non-surgical management. Four children (one with extensive tracheo-bronchomalacia and one with CHARGE association) had anti-reflux surgery (three Nissen and one Thal). We believe that oesophageal tailoring and augmentation for reconstruction of the oesophagus has the advantage of creating a more uniform oesophagus thus avoiding swallowing difficulty, bolus obstruction and the need of oesophageal dilatations too often accepted as integral to the problem following OA repair. PMID- 16344935 TI - Amyand's hernia presenting as neonatal testicular ischaemia. AB - Testicular ischaemia presenting in the neonatal period is most often attributable to neonatal torsion. We present an unusual case of a male neonate who presented with acute appendicitis within a patent processus vaginalis, causing cord compression and consequent testicular ischaemia. PMID- 16344936 TI - Effects of pressure exerted on the skin by elastic cord on the core temperature, body weight loss and salivary secretion rate at 35 degrees C. AB - Effects of pressure exerted on the skin by elastic cord on the core temperature, body weight loss and salivary secretion rate were studied under conditions of ambient temperature of 35 degrees C and a relative humidity of 60%. Twelve healthy females, aged 18-23 years, served as subjects. The subjects entered a bioclimatic chamber and rested quietly in a chair for 80 min. Then, skin pressure was exerted by applying elastic cord (8.5 mm wide) to six different skin areas, such as axilla, under-bust, waist, inguines, thighs and ankles. The values of skin pressure by elastic cord ranged from 11.9 to 33.3 g/cm(2). In the control experiment, wrapping with an elastic cord was loosely performed without any skin pressure. Rectal and skin temperatures, body weight loss by sweating and salivary secretion rate were measured throughout the 160 min experimental period. Core temperature increased more significantly under pressure exerted on the skin. Body weight loss by mainly sweating and salivary secretion rate were significantly suppressed under pressure exerted on the skin. We discussed the physiological mechanisms in terms of suppression of central nervous activity as to why significant increase of core temperatures, inhibition of body weight loss mainly by sweating and of salivary secretion rate occurred, and furthermore practical significance of these findings for impairment of digestion, swallowing, vocalizing, defense against disease bacteria and sport activity. PMID- 16344937 TI - Minor changes in blood lipids after 6 weeks of high-volume low- intensity physical activity with strict energy balance control. AB - Physical activity has been shown to favorably affect metabolic risk markers, including blood lipids. The impact of high-energy turnover, without influencing the traditionally used markers of exercise training effects, on blood lipids is still unclear. The aim was to study the effect of high-volume low-intensity physical activity on the blood lipid pattern, with a tight control of diet and energy balance. Eight untrained men [42.5 (12.1) years, body mass index 24.2 (2.8) kg m(-2)] were tested in two different 6-week protocols. In the sedentary protocol, the subjects were instructed to limit their everyday physical activity. In the activity protocol, a 2-h physical activity bout was performed 5 days week( 1) (approximately 40% of VO(2max); equivalent of an additional 21 MJ week(-1) in energy expenditure). The diet for both protocols comprised approximately 40 energy percent (E%) fat, approximately 50 E% carbohydrates (CHO). The polyunsaturated fatty acids to saturated fatty acids ratio of the diet was approximately 0.12. There were no changes during each 6-week period or differences between the two protocols in body weight, body composition or aerobic capacity. Low-intensity physical activity did not affect lipid parameters substantially, except for a slightly lower Apo-B/Apo-A1 ratio with the activity protocol (P < 0.05). Total and low-density lipoprotein cholesterol, as well as Apo-B and Apo-A1, were increased during the beginning of each 6-week period (P < 0.05), but returned to basal levels by the sixth week. In conclusion, 6 weeks of high-volume low-intensity physical activity did not affect blood lipids substantially. PMID- 16344938 TI - Effect of combined movement and heart rate monitor placement on physical activity estimates during treadmill locomotion and free-living. AB - A placement effect on activity measures from movement sensors has been reported during treadmill and free-living activity. Positioning of electrodes may impact on movement artifact susceptibility as well as surface ECG waveform amplitudes and thus potentially on the precision by which heart rate (HR) is ascertained from such ECG traces. The purpose of this study was to examine the extent to which placement of the combined HR and movement sensor, Actiheart, influences measurement of HR and movement, and estimates of energy expenditure. A total of 24 participants (20-39 years, 45-109 kg, 1.54-2.05 m, 19-29 kg m(-2)) were recruited. Whilst wearing two monitors, one placed at the level of the third intercostal space (upper position) and one just below the apex of the sternum (lower position), study participants performed level walking, incline walking, and level running on treadmill, and completed at least one day of free-living monitoring. Placement differences in HR data quality, movement counts, and energy expenditure (estimated from combined HR and movement) were analyzed with regression techniques. Quality of HR data was generally higher when monitors were placed in the lower position. This effect was more pronounced in men during both treadmill activity (relative risk, RR [95% CI] of noisy HR data in upper vs. lower position, RR=1.3[0.3; 5.6] in women, RR=174[14; 2,156] in men) and during free-living (RR=1.2[0.4; 3.3] in women, RR=25[9.6; 67] in men). There were minor placement differences (< or =8%) in movement counts only in women during incline walking and running. During free-living, no placement effect on counts was observed. In all test scenarios, estimates of energy expenditure from the two positions were not significantly different. Positioning the Actiheart at the level below the sternum may yield cleaner HR data. Regardless of which position is used, this has little or no effect on movement counts and energy expenditure estimates, which is encouraging for studies where research participants may have to position the monitors themselves. PMID- 16344939 TI - Cardiac autonomic dysfunction in rats chronically treated with anabolic steroid. AB - To date no published data exist regarding the effects of chronic high-dose anabolic-androgenic steroid administration on tonic cardiac autonomic control. The aim of this study was to evaluate, by power spectral analysis of heart rate variability (HRV), the effects of chronic treatment with supraphysiological doses of nandrolone decanoate (DECA) on tonic cardiac autonomic regulation in sedentary rats. Male Wistar rats were treated weekly with 10 mg kg(-1) of DECA (n=7) or vehicle (CONTROL, n=7) for 10 weeks. At the 8th week of treatment, electrocardiogram was recorded in the conscious state, for time- and frequency domain HRV analysis. Parasympathetic indexes were reduced in DECA group: high frequency power (CONTROL=11.1+/-3.0 ms2 vs. DECA=3.8+/-0.6 ms2, P<0.05), RMSSD (CONTROL=5.9+/-0.9 ms vs. DECA 3.5+/-0.3 ms; P<0.05) and pNN5 (CONTROL=31.5+/-7.5 ms vs. DECA=13.2+/-2.6 ms; P<0.05). The sympathetic index LF/HF tended to be higher in DECA group (CONTROL=0.65+/-0.15 vs. DECA=1.17+/-0.26, P=0.0546). In conclusion, chronic treatment with DECA, in rats, impairs tonic cardiac autonomic regulation, which may provide a key mechanism for anabolic steroid-induced arrhythmia and sudden cardiac death. PMID- 16344940 TI - The physiological response of ankle systolic blood pressure and ankle to brachial index after maximal exercise in athletes is dependent on age. AB - The development of sports activities in the over forties has increased the number of middle-aged patients performing heavy-load exercise. The normal response of ankle systolic blood pressure (ASBP) and ankle to brachial index (ABI) to intense exercise is known in young athletes, but little is known for other age groups. Three groups of 21 athletes: <20 (G1), 20-40 (G2), and >40 (G3) years old, respectively, were studied. ASBP and brachial systolic blood pressure (BSBP) were measured simultaneously before and after an incremental maximal cycle ergometer test. Rest ABI was higher in G3 compared to G1 (P<0.001: ANOVA, Bonferroni). Compared to respective resting values, BSBP, at 1 min of recovery, was increased for the three groups of age (P<0.001) whereas ASBP was unchanged for G1 and G2, and significantly increased for G3 (P<0.001). ASBP was significantly higher in G3 than in G1 and G2, ABI was lower in G1 and G2 compared to G3: 0.70+/-0.11, 0.76+/ 0.09 and 0.91+/-0.14, respectively. This age-related ABI increase is consistent with the impaired vasodilator capacity observed in aging normal subjects but not with the decrease observed in ABI resting values in epidemiological studies. ABI is higher in older than in younger athletes at rest and after exercise. In aging athletes, aiming to detect mild to moderate arterial lesions, it is likely that normal limits defined in young athletes are not valid. PMID- 16344941 TI - Plasma oxidative stress biomarkers, nitric oxide and heat shock protein 70 in trained elite soccer players. AB - The physiological response to the physical exercise involves a number of changes in the oxidative balance and in the metabolism of some important biological molecules, including nitric oxide (NO) and heat shock proteins (Hsp 70). With the aim to optimise previous laboratory diagnostic panels, we measured the plasma concentration of reactive oxygen metabolites (ROMs), total antioxidant status (TAS), glutathione reductase (GR) activity, and NO and Hsp 70 levels in 44 elite, antioxidant-supplemented and trained soccer players and in 15 sedentary controls. Although no statistically significant difference between athletes and controls was detected in the plasma level of ROMs and TAS, soccer players showed a significantly higher plasma GR activity, NO and Hst 70 levels than those of sedentary controls. These findings suggest that the measuring of relatively novel biomarkers in sport medicine, like GR, NO and Hsp 70, in addition to the well known and reliable assays (d-ROMs test and TAS) may be useful to a clinician to better assess and evaluate the benefits of training and/or supplementation programs. PMID- 16344942 TI - Effect of quadriceps contraction on upper limb position sense errors in humans. AB - Short-length muscle contraction (hold-short conditioning) causes limb position sense errors after the muscle returns to its intermediate length; this is due to intrafusal muscle thixotropy, which raises the muscle spindle sensitivity. In humans, contraction of muscles in the upper body (referred to as the Jendrassik manoeuvre) reinforces tendon reflexes in the lower limbs. However, it is unclear whether such a reinforcement manoeuvre enhances thixotropic position sense errors. We studied the effect of quadriceps contraction on upper limb position sense errors induced by hold-short conditioning of the biceps in 12 healthy men. Quadriceps contraction increased the tonic vibration reflex of the biceps, suggesting that quadriceps contraction has a reinforcing effect similar to that of the Jendrassik manoeuvre. After hold-short conditioning of the right biceps, subjects perceived that the conditioned forearm was placed in a more extended position than it actually was. Such position sense errors were enhanced during quadriceps contraction and the degree of error was increased with the intensity of the quadriceps contraction. These results suggest that limb position sense is affected by remote muscle contraction. PMID- 16344943 TI - Reproducibility of relationships between human ventilation, its components and oesophageal temperature during incremental exercise. AB - For human exercise at intensities greater than approximately 70 to 85% of maximal levels of exertion, ventilation (V E) increases proportionately to core temperature (T C) following distinct T C thresholds. This suggested T C in humans could be a modulator of exercise-induced ventilation. This study tested the reproducibility of relationships between oesophageal temperature (T oes) ventilation and its components during incremental exercise. On two nonconsecutive days, at an ambient temperature of 22.1+/-0.3 degrees C and RH of 45+/-5%, seven untrained adult males of normal physique pedaled on a seated cycle ergometer in an incremental exercise protocol from rest to the point of exhaustion. In each exercise session, ventilatory equivalents for oxygen consumption (VE.VO2 (1-)) and carbon dioxide production (VE.VCO2 (1-)) plus the components of V E, tidal volume (V T) and frequency of respiration (f), were expressed as a function of T oes. Results indicated the reproducibility criteria of Bland and Altman were met for the relationships between T oes and both (VE.VO2 (1-)) and (VE.VCO2 (1-)) as well as for relationships between T oes and each of V T and f. Intraclass correlation coefficients (R) for between-trial T oes thresholds for (R=0.91, P<0.05) and (R=0.88, P<0.05) were also high and significant. In both trials, after T oes increased by approximately 0.3 degrees C, V T demonstrated a distinct plateau point at a reproducible T oes (R=0.93, P<0.05) and f demonstrated a distinct and reproducible T oes threshold (R=0.84, P<0.05). In conclusion, the results illustrate that for humans, ventilation has a significant and reproducible relationship with core temperature during incremental exercise. PMID- 16344944 TI - Adjustment of the human arm viscoelastic properties to the direction of reaching. AB - The viscoelastic properties of the human arm were measured by means of short force perturbations during fast reaching movements in two orthogonal directions. A linear spring model with time delay described the neuromuscular system of the human arm. The obtained viscoelastic parameters ensured movement stability in spite of the time delay of 50 ms. The stiffness and viscosity ellipses appeared to be predominantly orthogonal to the movement direction, which reduced the effect of force perturbation in the direction orthogonal to the reaching movement. Thus, it can be argued that the viscoelastic properties of the neuromuscular system of the human arm are adjusted to the direction of movement according to a "path preserving" strategy, which minimizes the deviation of the movement path from a straight line, when exposed to an unexpected external force. PMID- 16344946 TI - Annexins as cell-type-specific markers in the developing chicken chorionallantoic membrane. AB - Between day E8 and E12 of embryonic development, the chicken chorioallantoic membrane (CAM) undergoes massive structural rearrangement enabling calcium-uptake from the eggshell to supply the growing embryo. However, the contribution of the various cell types of the chorionic epithelium including the capillary covering (CC) cells, villus cavity (VC) cells, endothelial-like cells, and basal cells to this developmental program is largely unknown. In order to obtain markers for the different cell types in the chorionic epithelium, we determined the expression patterns of various calcium-binding annexins in the developing chicken CAM. By reverse transcription/polymerase chain reaction with primers deduced from nucleotide sequences available in various databases, the presence of annexin (anx)-1, anx-2, anx-5, and anx-6 was demonstrated at days E8 and E12. Quantitative immunoblotting with novel antibodies raised against the recombinant proteins revealed that anx-1 and anx-5 were significantly up-regulated at day E12, whereas anx-2 and anx-6 expression remained almost unchanged in comparison to levels at day E8. Immunohistochemistry of paraffin-embedded sections of E12 CAM revealed anx-1 in CC cells and VC cells. Anx-2 was localized in capillaries in the chorionic epithelium and in basal cells of the allantoic epithelium, whereas anx-6 was detected in basal cells or endothelial-like cells of the chorionic epithelium and in the media of larger vessels in the mesenchyme. A 2 day exposure of the CAM to a tumor cell spheroid resulted in strong proliferation of anx-1-expressing CC cells suggesting that these cells participate in the embryonic response to experimental intervention. Thus, annexins exhibit complementary expression patterns and represent appropriate cell markers for the further characterization of CAM development and the interpretation of results obtained when using CAM as an experimental model. PMID- 16344945 TI - A hot pepper gene encoding WRKY transcription factor is induced during hypersensitive response to Tobacco mosaic virus and Xanthomonas campestris. AB - Plant WRKY transcription factors were previously implicated in the alteration of gene expression in response to various pathogens. The WRKY proteins constitute a large family of plant transcription factors, whose precise functions have yet to be elucidated. Using a domain-specific differential display procedure, we isolated a WRKY gene, which is rapidly induced during an incompatible interaction between hot pepper and Tobacco mosaic virus (TMV) or Xanthomonas campestris pv . vesicatoria (Xcv). The full-length cDNA of CaWRKY-a (Capsicum annuum WRKY-a) encodes a putative polypeptide of 546 amino acids, containing two WRKY domains with a zinc finger motif. The expression of CaWRKY-a could be rapidly induced by not only chemical elicitor such as salicylic acid (SA) or ethephon but also wounding treatments. The nuclear localization of CaWRKY-a was determined in transient expression system using tobacco BY-2 cells by polyethylene glycol (PEG) mediated transformation experiment. With oligonucleotide molecules containing the putative W-box sequences as a probe, we confirmed that CaWRKY-a protein had W-box binding activity. These results suggest that CaWRKY-a might be involved as a transcription factor in plant defense-related signal transduction pathways. PMID- 16344947 TI - Down-regulation of vasoactive intestinal peptide and altered expression of its receptors in rat diabetic cardiomyopathy. AB - Vasoactive intestinal peptide (VIP) is a vasorelaxant peptide that addresses two receptor subtypes, VPAC1 and VPAC2. It stimulates insulin secretion and mediates anti-inflammatory effects and has been proposed for treatment of type 2 and autoimmune diabetes. In the heart, VIP is produced and released primarily by intrinsic neurons and improves cardiac perfusion and function. Here, we investigated the involvement of this system in the events underlying development of experimentally induced diabetic cardiomyopathy. Rats received a single streptozotocin injection, and cardiac VIP content [radioimmune assay (RIA)], expression of the VIP precursors VPAC1 and VPAC2 [real-time reverse transcription polymerase chain reaction (RT-PCR)], and VPAC1 and VPAC2 tissue distribution (immunohistochemistry) were assessed 4, 8, and 16 weeks thereafter and compared with corresponding vehicle-treated controls. Cardiac neuropathy manifests progressively during the first 4 months of diabetes at the preproVIP mRNA and VIP peptide level and is accompanied by initial down-regulation of VPAC2 at one prime target of VIP-containing axons, i.e., smooth muscle cells of coronary arterioles. VPAC1 is expressed by macrophages. After initial changes that are specific for atria and ventricles, respectively, VPAC1 and VPAC2 expression return to control levels at 16 weeks despite ongoing loss of VIP. Given the cardioprotective role of the VIP signaling system, the persistence of receptors has therapeutic implications since it is the prerequisite for trials with VPAC2 agonists. PMID- 16344948 TI - The environment of amide groups in protein-ligand complexes: H-bonds and beyond. AB - A comprehensive structural analysis of interactions involving amide NH and C=O groups in protein-ligand complexes has been performed based on 3,275 published crystal structures (resolution < or =2.5 A). Most of the amide C=O and NH groups at the protein-ligand interface are highly buried within the binding site and involved in H-bonds with corresponding counter-groups. Small percentages of C=O and NH groups are solvated or embedded in hydrophobic environments. In particular, C=O groups show a higher propensity to be solvated or embedded in a hydrophobic environment than NH groups do. A small percentage of carbonyl groups is involved in weak hydrogen bonds with CH. Cases of dipolar interactions, involving carbonyl oxygen and electrophilic carbon atoms, such as amide, amidinium, guanidium groups, are also identified. A higher percentage of NH are in contact with aromatic carbons, interacting either through hydrogen bonds (preferably with the NH group pointing towards a ring carbon atom) or through stacking between amide plane and ring plane. Comprehensive studies such as the present one are thought to be important for future improvements in the molecular design area, in particular for the development of new scoring functions. [Figure: see text]. PMID- 16344949 TI - Modeling study of the influences of the aromatic transitions and the local environment on the far-UV rotational strengths in TEM-1 beta-lactamase. AB - Rotational strengths in the far-UV of TEM-1 beta-lactamase have been investigated with two theoretical models based on the matrix method. The first model excludes, and a second includes, effects of the local electrostatic interactions on the chromophore energies. Special attention is given to the contributions of the aromatic side-chain chromophores, and the mechanisms of generation of rotational strengths are analyzed. The sensitivity of the computational models with respect to the structural changes of the protein are discussed. [Figure: see text]. PMID- 16344950 TI - Application of artificial neural networks for predicting the aqueous acidity of various phenols using QSAR. AB - Artificial neural networks (ANNs) have been successfully trained to model and predict the acidity constants (pK(a)) of 128 various phenols with diverse chemical structures using a quantitative structure-activity relationship. An ANN with 6-14-1 architecture was generated using six molecular descriptors that appear in the multi-parameter linear regression (MLR) model. The polarizability term (pi (I)), most positive charge of acidic hydrogen atom (q+), molecular weight (MW), most negative charge of the phenolic oxygen atom (q-), the hydrogen bond accepting ability (epsilon(B)) and partial-charge weighted topological electronic (PCWTE) descriptors are inputs and its output is pK(a). It was found that a properly selected and trained neural network with 106 phenols could represent the dependence of the acidity constant on molecular descriptors fairly well. For evaluation of the predictive power of the ANN, an optimized network was used to predict the pK(a)s of 22 compounds in the prediction set, which were not used in the optimization procedure. A squared correlation coefficient (R2) and root mean square error (RMSE) of 0.8950 and 0.5621 for the prediction set by the MLR model should be compared with the values of 0.99996 and 0.0114 by the ANN model. These improvements are due to the fact that the pK(a) of phenols shows non linear correlations with the molecular descriptors. [Figure: see text]. PMID- 16344951 TI - [Pharmacotherapy of supraventricular arrhythmias. What comes, what remains, what goes?]. AB - Supraventricular tachycardias consist of AV-nodal-reentrant-tachycardias, atrioventricular tachycardias with accessory pathways (WPW-syndrome), atrial tachycardias, atrial fibrillation and atrial flutter. Only specific ECG interpretation with an exact arrhythmia classification offers the way to perform modern differential therapy including drug treatment and also interventional therapy modalities. In atrial fibrillation, drug treatment is still first-line therapy: physicians have to make a decision either to follow the rate or rhythm control concept. In case of rhythm control, drug therapy is tailored to the individual patient taking into account the patients symptomatology, left ventricular ejection fraction and nature and degree of an underlying cardiac disease. Drug refractory symptomatic atrial fibrillation patients should be considered for interventional treatment like pulmonary vein ablation. Recurrent typical right atrial flutter, AV-nodal-reentrant-tachycardia and all forms of atrioventricular tachycardias however are indications for catheter ablation; long term drug treatment will only be performed in rare cases. PMID- 16344952 TI - [Amiodarone or ICD in patients with cardiac insufficiency. SCD-HeFT Study]. PMID- 16344953 TI - [Soft tissue defects as a complication in knee arthroplasty. Surgical strategies for soft tissue reconstruction]. AB - Soft tissue defects after knee arthroplasty are a severe problem, which can even result in a loss of the prosthesis or the limb. Well-planned strategies are necessary for sufficient soft tissue reconstruction, resulting in optimal functional and aesthetic results. This report provides information on the classification of the tissue defects and the appropriate options for surgical reconstruction. Besides the basic therapy principles of immobilisation, debridement, planned lavages and antibiotic therapy, defect-dependent surgical techniques of reconstructive surgery are implemented. These include skin transplantation, local fasciocutaneous flaps, local pedicled muscle flaps and free flaps. For best results, interdisciplinary treatment by orthopaedic surgeons, plastic surgeons, microbiologists and physiotherapists is mandatory. PMID- 16344954 TI - [Necrosis of the jaws under bisphosphonate therapy]. AB - Bisphosphonates are widely used in the treatment of cancer patients with hypercalcemia and bone metastases or in osteoporosis therapy. Current reports have focused on therapy-resistant osteonecrosis of the jaws as a possible side effect of bisphosphonates. Official German drug committees have recently warned about the possibility of these side effects. So far we have experience with 12 patients showing therapy-resistant osteonecrosis of the mandible under bisphosphonate medication, three of whom received oral bisphosphonates for osteoporosis treatment. Presentation of these three cases provides more information on this clinically important side effect of oral bisphosphonate medication, also in osteoporosis therapy. PMID- 16344955 TI - [Improving the diagnosis of septic arthritis by use of a pediatric blood culture system]. AB - BACKGROUND: Prognosis and outcome of bacterial joint infections are dependent on the fast and reliable identification of pathogens in the synovial fluid. Previous studies have suggested the possible advantage of using a blood culture system in contrast to conventional culture methods. PATIENTS AND METHODS: A total of 101 synovial specimens from patients presenting with symptoms suggesting septic arthritis were taken by aspiration with a sterile syringe. We compared the diagnostic results of automated analysis in a blood culture system against conventional culture on solid agar plates. RESULTS: Some 67 specimens (66.3%) were found to be negative in both preparations, while samples from 21 patients (20.8%) yielded the same microorganisms. In 13 cases (12.9%), the isolation of a pathogen was possible only with the blood culture method, whereas the conventional method never yielded a positive result when the blood culture was negative. Thus, the diagnostic yield was significantly improved by use of the blood culture system (P <0.001). CONCLUSION: The use of a commonly available blood culture system offers a fast, reliable and cost-effective approach for the diagnosis of septic arthritis and should therefore be considered as an useful alternative to conventional culture methods. PMID- 16344957 TI - Interaction between grounding pads used for RF ablation therapy and magnetic resonance imaging. AB - OBJECTIVES: To characterize artifacts and imaging problems in the presence of conductive grounding pads for RF ablation therapy as well as potential heating problems due to induction of eddy currents in the pads. Strategies for avoidance of those problems are developed. MATERIALS AND METHODS: Underlying principles of interactions between grounding pads and MR imaging are reported. Influential parameters, e.g., orientation in relation to the magnetic field, shape of the grounding pad, sequence type (spin-echo versus gradient echo) and magnetic field strength (0.2 T, 1.5 T, 3 T) were varied in systematic phantom studies. Heating effects due to induced eddy currents were estimated theoretically and measured by infrared imaging in an adapted set-up. RESULTS: MR imaging artifacts are markedly dependent on the orientation and geometrical shape of the grounding pads. Visible signal extinction artifacts were more pronounced using spin-echo techniques than in gradient echo images and increased for higher field strengths. Suitable incisions in the grounding pad reduced eddy currents markedly and minimized image artifacts. Heating problems due to induced eddy currents by the RF transmitted for MR imaging were excluded by phantom measurements. CONCLUSIONS: Suitable positioning of the grounding pads and adaptation of their geometry provide clearly reduced artifacts in MR imaging. PMID- 16344956 TI - Whole genome expression profiling of the medial and lateral substantia nigra in Parkinson's disease. AB - We have used brain tissue from clinically well-documented and neuropathologically confirmed cases of sporadic Parkinson's disease to establish the transcriptomic expression profile of the medial and lateral substantia nigra. In addition, the superior frontal cortex was analyzed in a subset of the same cases. DNA oligonucleotide microarrays were employed, which provide whole human genome coverage. A total of 570 genes were found to be differentially regulated at a high level of significance. A large number of differentially regulated expressed sequence tags were also identified. Levels of mRNA sequences encoded by genes of key interest were validated by means of quantitative real-time polymerase chain reaction (PCR). Comparing three different normalization procedures, results based on the recently published GeneChip Robust Multi Array algorithm were found to be the most accurate predictor of real-time PCR results. Several new candidate genes which map to PARK loci are reported. In addition, the DNAJ family of chaperones is discussed in the context of Parkinson's disease pathogenesis. PMID- 16344958 TI - A simple method for mapping the B1 field distribution of linear RF coils. AB - Inhomogeneity of the radio frequency (RF) field B1 leads to intensity variations in MR images and to spatial dependence of spectral line amplitudes. In this paper, a simple method of measuring the B1 field components of an unsegmented linear coil is described. The method is designed for the coils operating up to 20 MHz. The B1 field distribution is replaced by the static magnetic field caused by DC current flowing through the coil. The technique involves rotating the coil 90 degrees so that measured B1 component is aligned with B0 and measuring the shift of resonance frequency using a spectroscopic imaging sequence. Experimental results were in good agreement with the theoretical computations. PMID- 16344959 TI - NADPH oxidases in Eukaryotes: red algae provide new hints! AB - The red macro-alga Chondrus crispus is known to produce superoxide radicals in response to cell-free extracts of its green algal pathogenic endophyte Acrochaete operculata. So far, no enzymes involved in this metabolism have been isolated from red algae. We report here the isolation of a gene encoding a homologue of the respiratory burst oxidase gp91(phox) in C. crispus, named Ccrboh. This single copy gene encodes a polypeptide of 825 amino acids. Search performed in available genome and EST algal databases identified sequences showing common features of NADPH oxidases in other algae such as the red unicellular Cyanidioschyzon merolae, the economically valuable red macro-alga Porphyra yezoensis and the two diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. Domain organization and phylogenetic relationships with plant, animal, fungal and algal NADPH oxidase homologues were analyzed. Transcription analysis of the C. crispus gene revealed that it was over-transcribed during infection of C. crispus gametophyte by the endophyte A. operculata, and after incubation in presence of atrazine, methyl jasmonate and hydroxyperoxides derived from C20 polyunsaturated fatty acids (PUFAs). These results also illustrate the interest of exploring the red algal lineage for gaining insight into the deep evolution of NADPH oxidases in Eukaryotes. PMID- 16344960 TI - Cadmium inducible Fe deficiency responses observed from macro and molecular views in tobacco plants. AB - Responses induced by Cd exposure were assessed in tobacco seedlings (Nicotiana tabacum L.) using macro and molecular indices. The 100 microM of Cd exposure reduced the total dry weight and chlorophyll index of the seedlings as much as the genuine Fe-deficiency. Concentration of Fe in the shoots decreased, whereas that in the roots increased by the Cd exposure, especially in the apoplasmic space. It is probable that Cd interferes mainly with the step of Fe-translocation from the roots to shoots and this sets the upper-part of the plant in a state of Fe-deficiency. The Cd exposure coordinately increased the expressions of the exogenous and the endogenous Fe-deficiency responsive genes, HvIDS2(pro)::GUS, NtFRO1 and NtIRT1 in the roots. This is the first data to demonstrate the responses of Cd-inducible Fe-deficiency at a molecular level. PMID- 16344961 TI - MR-guided biopsies with a newly designed cordless coil in an open low-field system: initial findings. AB - The purpose of this study was to examine the feasibility and safety of MR-guided biopsies with a newly designed cordless coil in an open low-field magnetic resonance (MR) system. Eleven patients were biopsied using a low-field system (0.2 T, Magnetom Concerto, Siemens) by using the new cordless coil (Siemens). The biopsies were performed in different organ systems [liver (n = 7), abdomen (n = 1), shoulder (n = 1), pelvis (n = 1) and hip (n = 1)]. The procedures were guided using T1-weighted FLASH (fast low-angle shot) sequences. The lesions were biopsied using the coaxial technique through a 15-gauge puncture needle with a 16 gauge biopsy handy. Coil handling, image quality and complications were evaluated. Imaging quality and visualization of the lesions were optimal up to a penetration depth of 9 cm. In all cases the biopsy procedures were successfully performed with MR guidance without any complications. Pathological findings revealed seven cases of malignant tissue and four cases of non-malignant tissue. The use of the cordless coil allows improved patient access during the biopsy and an improved handling of the coil system. MR-guided biopsy using the novel cordless coil system can be performed safely and precisely with easy handling of the coil. This coil concept, however, is restricted to special indications. PMID- 16344962 TI - Development of water-soluble metallofullerenes as X-ray contrast media. AB - Fullerenes are a new carbonic allotrope having a cage structure. We investigated whether fullerenes containing one or two atoms of heavy metals could be an X-ray contrast material with little adverse effects. One or two atoms of dysprosium (Dy), erbium (Er), gadolinium (Gd), europium (Eu) and lutetium (Lu) were encapsulated into fullerene (C(82)), which was synthesized as a polyhydroxyl form (e.g., Gd@C(82)(OH)n, n=40, Gd - fullerenols). They were dissolved in water at maximum soluble concentrations and subjected to CT number analysis. The CT numbers of the solutions were measured using a 4- or 16-row multidetector CT scanner. The CT number of the water-soluble metallofullerenes were 56.0 HU for Dy@C(82)(OH)(40), 111.5 HU for Er@C(82)(OH)(40), 58.4 HU for Gd@C(82)(OH)(40), 100.9 HU for Eu@C(82)(OH)(40) and 23.3 HU for Lu(2)@C(82)(OH)(40). The CT numbers of the metallofullerenes investigated in the present study were not high enough to be used in the clinic in place of iodinated contrast materials. However, if nanotechnology progresses in the near future, it may prove to have a possibility as an X-ray contrast material. PMID- 16344964 TI - Multifocal arterial haemorrhage in a partially stable pelvic fracture after a crush injury: a case report. AB - INTRODUCTION: Most pelvic haemorrhages are thought to be caused by injury to small arteries or veins in the fractured cancellous pelvic bone or in the surrounding soft-tissues, and only 6-18% of patients with unstable pelvic fractures have haemorrhage from larger arteries. When arterial injuries are present, the majority involve branches of the internal iliac artery with only few published reports of injuries to the external iliac artery or its branches. MATERIALS AND METHODS: We report of a patient who sustained a combined pelvic and acetabular fracture with multifocal bleeding involving branches of both the internal iliac as well as the external iliac arteries after a crush injury. The primary attention was focused on the most probable arterial injury, the internal iliac artery and only at repeat angiography was the injury to the internal epigastric artery, caused by degloving injury to the trunk, recognized. RESULTS: Arterial control was achieved only following aggressive fluid resuscitation, pelvic packing, repeated embolization and ligation of the peripelvic inferior epigastric artery. After initial haemodynamic control was achieved, the patient sustained multiple complications, partly as a consequence of the injury, but also as a consequence of the life-saving treatments. CONCLUSION: The case describes a rare combination of arterial injuries in a complex pelvic fracture constituting a partially stable pelvic fracture and a dislocated acetabular fracture in a patient with pelvic crush injury and a degloving injury to the trunk. The case also describes the complex nature of these injuries and rarely reported problems related to the treatment of them. PMID- 16344965 TI - How did the platypus get its sex chromosome chain? A comparison of meiotic multiples and sex chromosomes in plants and animals. AB - The duck-billed platypus is an extraordinary mammal. Its chromosome complement is no less extraordinary, for it includes a system in which ten sex chromosomes form an extensive meiotic chain in males. Such meiotic multiples are unprecedented in vertebrates but occur sporadically in plant and invertebrate species. In this paper, we review the evolution and formation of meiotic multiples in plants and invertebrates to try to gain insights into the origin of the platypus meiotic multiple. We describe the meiotic hurdles that translocated mammalian chromosomes face, which make longer chains disadvantageous in mammals, and we discuss how sex chromosomes and dosage compensation might have affected the evolution of sex linked meiotic multiples. We conclude that the evolutionary conservation of the chain in monotremes, the structural properties of the translocated chromosomes and the highly accurate segregation at meiosis make the platypus system remarkably different from meiotic multiples in other species. We discuss alternative evolutionary models, which fall broadly into two categories: either the chain is the result of a sequence of translocation events from an ancestral pair of sex chromosomes (Model I) or the entire chain came into being at once by hybridization of two populations with different chromosomal rearrangements sharing monobrachial homology (Model II). PMID- 16344967 TI - DXS10079, DXS10074 and DXS10075 are STRs located within a 280-kb region of Xq12 and provide stable haplotypes useful for complex kinship cases. AB - The evaluation of the short tandem repeat (STR) markers DXS10079, DXS10074 and DXS10075 was amended to establish a STR cluster spanning a genetic distance<1 cM. These three STRs are located within a 280-kb region at Xq12 and provide stable haplotypes useful for solving complex kinship cases. Theoretically, this cluster could give rise to 2,548 different haplotypes in the German population and the genotyping of 781 men revealed the presence of 172 haplotypes. Since the three STRs were shown to be in strong linkage disequilibrium (LD), haplotype frequencies cannot be computed on the basis of a single locus allele frequency alone but have to be estimated directly. Here, we present data on linkage, haplotype frequencies and LD in a German population. Further clusters from other regions of the X chromosome will be published in the future to cover the chromosome with a well-structured network of highly informative sites. PMID- 16344969 TI - Explaining bank vole cycles in southern Norway 1980-2004 from bilberry reports 1932-1977 and climate. AB - Correlations between mast fruiting of bilberry Vaccinium myrtillus and peak levels of Clethrionomys-voles have been reported from both Norway and Finland, but there has been a discussion whether this is a bottom-up or a top-down relationship. In a multiple regression model, 65% of the variation in a bilberry production index calculated from game reports from southern Norway 1932-1977 could be explained by the berry index of the two preceding years and climate factors acting during key stages of the flowering cycle. High vole populations in previous years did not contribute to explain the fluctuation in berry production. I used the selected model and climate data to predict bilberry production for the period 1978-2004. Predicted berry indices of the current and previous year explained 38% and the total amount of precipitation in May-June explained 16% of the variation in a log-transformed snap-trapping index of bank vole Clethrionomys glareolus 1980-2004. The vole index was not related to any of the climate variables used to predict berry production. This pattern supports the hypothesis that vole cycles are generated by changes in plant chemistry due to climate synchronized mast fruiting. PMID- 16344968 TI - Medaka double mutants for color interfere and leucophore free: characterization of the xanthophore-somatolactin relationship using the leucophore free gene. AB - Somatolactin (SL) plays an essential role in body-color regulation in medaka and is encoded by the color interfere (ci) locus. The ci mutant fish possess constitutively increased numbers of leucophores and a concomitant decrease in visible xanthophores. However, the mechanism of action of SL on these cell types, and the role of SL in body-color regulation in other species, is unknown. In this study, we verified an SL-xanthophore relationship in ci mutant fish using the leucophore free (lf) gene. Histological observation of lf larvae indicated that these mutants do not possess differentiated leucophores. The ci-lf double mutant, whose genotype was confirmed using DNA markers, lacked leucophores; however, the number of xanthophores remained low, demonstrating that leucophores are not necessary for mediating SL signaling to xanthophores. This finding suggests a conserved function for SL in xanthophore regulation across species, rather than the evolution of a medaka-specific and leucophore-dependent role of SL in body color regulation. Our results also demonstrate that the lf gene has an indispensable role in leucophore development epistatic to SL signaling. The lf gene has not been cloned. The high-resolution recombination map surrounding the lf locus constructed in this study, together with medaka whole genome sequences that will be released soon, will allow the rapid cloning of the lf gene by forward genetic approaches. PMID- 16344970 TI - Models for dioxygen activation by the CuB site of dopamine beta-monooxygenase and peptidylglycine alpha-hydroxylating monooxygenase. AB - On the basis of spectroscopic and crystallographic data for dopamine beta monooxygenase and peptidylglycine alpha-hydroxylating monooxygenase (PHM), a variety of ligand sets have been used to model the oxygen-binding Cu site in these enzymes. Calculations which employed a combination of density functional and multireference second-order perturbation theory methods provided insights into the optimal ligand set for supporting eta (1) superoxo coordination as seen in a crystal structure of a precatalytic Cu/O(2) complex for PHM (Prigge et al. in Science 304:864-867, 2004). Anionic ligand sets stabilized eta (2) dioxygen coordination and were found to lead to more peroxo-like Cu-O(2) complexes with relatively exergonic binding free energies, suggesting that these adducts may be unreactive towards substrates. Neutral ligand sets (including a set of two imidazoles and a thioether), on the other hand, energetically favored eta (1) dioxygen coordination and exhibited limited dioxygen reduction. Binding free energies for the 1:1 adducts with Cu supported by the neutral ligand sets were also higher than with their anionic counterparts. Deviations between the geometry and energetics of the most analogous models and the PHM crystal structures suggest that the protein environment influences the coordination geometry at the Cu(B) site and increases the lability of water bound to the preoxygenated reduced form. Another implication is that a neutral ligand set will be critical in biomimetic models in order to stabilize eta (1) dioxygen coordination. PMID- 16344971 TI - Identification of adhesions on CT in small-bowel obstruction. AB - Identification of adhesions on computed tomography (CT) in cases of small-bowel obstruction (SBO) is currently a diagnosis of exclusion. The purpose of this study is to examine whether the presence of findings suggestive of an extraluminal band can be used as a CT sign for adhesive SBO. CT scans of 142 patients with surgically proven SBO performed within 72 h of exploratory surgery were reviewed. The studies were evaluated for the cause of SBO and the presence of extraluminal bands. An extraluminal band was considered present if there was a change in the conformation of the transition zone, suggesting extraluminal compression of the bowel by a linear obstructive band. The presence of extraluminal bands in the area of the transition zone had a high positive predictive value for adhesive SBO. Of the 73 cases in which bands were present, SBO was due to adhesions in 52 instances, with a corresponding positive predictive value of 71% (95% confidence interval 0.60-0.80) and a p value of 0.008. The extraluminal band was 61% sensitive and 63% specific for adhesive SBO. The presence of an extraluminal band on CT in the area of the transition zone in cases of SBO correlates well with a diagnosis of SBO secondary to adhesions. In the absence of a source of SBO, the presence of an extraluminal band can serve as a helpful diagnostic adjunct for adhesive SBO. PMID- 16344972 TI - Low-grade (often multifocal) osteomyelitis (a diagnostic problem and/or a mimicker of tumor). PMID- 16344973 TI - Head CT evaluation in clinically stable motor vehicle collision patients. AB - PURPOSE: To assess the utilization of non-contrast head computed tomography (HCT) in clinically stable patients who were involved in motor vehicle collisions. Our goal is to identify patients in whom HCT may be safely excluded without altering clinical outcome. METHODS: Adult, front-seat passengers were retrospectively examined. All clinically stable patients involved in motor vehicle collisions and who subsequently underwent HCT were studied to determine the factors that increased the statistical likelihood of a negative examination. Seven factors relevant to passenger safety or clinical outcomes were measured. RESULTS: Eighteen of the 206 patients who met criteria for our study had positive HCT findings. Front-seat passengers were more likely to suffer traumatic head injury than drivers. Both Glasgow Coma Scale (GCS) and Injury Severity Score (ISS) individually had statistically significant correlations with positive HCT findings. Thirteen of the 18 patients had GCS scores of 13-15. Seatbelts, rollover collisions, gender, and airbag deployment demonstrated relatively decreased predictive values. CONCLUSION: The ISS emerged as the single most reliable predictor of positive HCT findings. Regarding multiple variable effects, the combinations of ISS and patient position or GCS and patient position are the most statistically significant clinical factors in identifying patients in whom HCT may be safely excluded. PMID- 16344974 TI - Helical and multislice CTA following penetrating trauma to the subclavian and axillary arteries (pictorial essay). AB - Penetrating injuries with resultant trauma to the subclavian and axillary arteries have traditionally been evaluated with direct contrast angiography. Physical examination has a low sensitivity for detection of vascular injuries in stable patients and surgical exploration is challenging. With advancements in CT technology, a less invasive and more rapid approach in evaluating arterial injury is now being utilized in many trauma centers. This article will depict the CT angiographic signs of subclavian and axillary artery injuries. PMID- 16344975 TI - Occipital condyle fractures: clinical presentation and imaging findings in 76 patients. AB - The aim was to assess the frequency and type of occipital condyle fractures in patients with significant trauma, and determine the frequency in which conventional radiographic findings are positive. Secondarily, we subjectively evaluate the application of existing classification systems. Fracture of the occipital condyle is an uncommon lesion that may be associated with craniocervical instability. Relying on conventional radiographs to detect these injuries may be inappropriate. An institutional trauma database of patients was searched for patients with occipital condyle fractures. The types of fractures were classified retrospectively based on re-review of imaging studies, using existing classification systems. Frequency of types was calculated, and the ease of use of the fracture classifications was evaluated subjectively. Conventional radiographs were reviewed for the presence of subjective soft tissue swelling and the visibility of the fracture(s). Seventy-six patients had CT images available for re-review. None of the occipital condyle fractures could be identified in the 60 patients who had radiographs available for re-review. Because of the presence of life support tubing and pharyngeal fluid limiting evaluation of prevertebral soft tissue swelling, the presence of widened prevertebral soft tissues was only helpful in 7 of the 60 patients. The multiplanar reformatted CT images were useful to determine alignment at C0-C1 and C1-C2. Occipital condyle fractures were not visualized on conventional radiographs. Secondary findings of soft tissue swelling were often absent or unreliable. CT scanning with multiplanar reconstruction imaging plays an indispensable role in evaluating for fractures of the cervical spine, and for determining alignment at C0-C1-C2. The most recently developed classification system of Tuli et al. (Neurosurgery 41: 368-376, 1997) is useful, but suffers from the lack of a defined distinction between undisplaced (Type I, stable) and displaced (Type 2A, unstable) fracture fragments. PMID- 16344976 TI - Clinical outcome of active extravasation in splenic trauma. AB - The purpose of this study was to determine the necessity for splenectomy in patients with active extravasation on contrast enhanced CT secondary to splenic trauma. We reviewed cases of splenic injury and classified these according to the American Association for the Surgery of Trauma (AAST) grading scale. The presence of active extravasation and associated injuries was assessed. Chart review was then performed to determine age, sex, mechanism of injury, indications for splenectomy, and clinical outcome. Of 82 cases evaluated, 12 grade I, 15 grade II, 30 grade III, 17 grade IV, and 8 grade V injuries were present. Eighteen patients were actively extravasating. Of extravasating patients, 13 eventually underwent open splenectomy or embolization and five (27.8%) were managed expectantly with success. Of grade IV injuries, 9/17 showed active extravasation, of which six underwent splenectomy. Of grade V injuries, 3/8 showed active extravasation, and all three underwent intervention. Splenectomy may not be necessary in appropriately chosen patients with active extravasation from the spleen in blunt abdominal trauma. PMID- 16344977 TI - CT directed diagnostic and therapeutic pericardiocentesis: 8-year experience at a single institution. AB - A retrospective review of the 8-year experience at a single institution performing more than 300 CT directed pericardiocenteses was performed. The technique, results, and complications were determined. Comparative data from the literature relating to cardiology and thoracic surgery are reviewed. A CPT code data base search was performed to identify all imaging directed pericardiocenteses at a 550 bed regional medical center between 1993 and 2001. Medical records, imaging studies, and reports as well as pertinent laboratory, chemistry and hematology values were reviewed. Demographics, results, fluid analyses, techniques, systems used and complications were recorded. RESULTS: Three hundred and nineteen pericardiocenteses using CT localization were attempted on 261 patients. One hundred and forty-two men were included and mean age was 61 years (range 6 months to 89 years). Mean volume aspirated was 418 cc (range 0-1,875 cc). The major complication rate was 0.3%; the minor complication rate was 6.9%. Technical success was achieved in 98.4% procedures. Emergent pericardiocentesis was performed on anticoagulated patients without significant bleeding. Seventy-nine percent of patients with echocardiographic evidence of tamponade had symptomatic improvement. Patients with marked shortness of breath without other evidence of tamponade benefited symptomatically in 78%. Pericardiocenteses with CT localization is relatively safe and many patients achieve symptomatic relief. PMID- 16344979 TI - Electronic publishing-a boon or a bugbear. PMID- 16344981 TI - Snake fangs from the Lower Miocene of Germany: evolutionary stability of perfect weapons. AB - There is a general consensus that most of today's nonvenomous snakes are descendants of venomous snakes that lost their venomous capabilities secondarily. This implies that the evolutionary history of venomous snakes and their venom apparatus should be older than the current evidence from the fossil record. We compared some of the oldest-known fossil snake fangs from the Lower Miocene of Germany with those of modern viperids and elapids and found their morphology to be indistinguishable from the modern forms. The primary function of recent elapid and viperid snake fangs is to facilitate the extremely rapid, stab-like application of highly toxic venoms. Our findings therefore indicate that the other components of the venom-delivery system of Early Miocene vipers and elapids were also highly developed, and that these snakes used their venom in the same way as their modern relatives. Thus, the fossil record supports the view that snakes used their venoms to rapidly subdue prey long before the mid-Tertiary onset of the global environmental changes that seem to have supported the successful radiation of venomous snakes. PMID- 16344978 TI - Retropharyngeal calcific tendinitis: case report and review of literature. AB - Retropharyngeal calcific tendinitis, also known as acute calcific prevertebral tendinitis or longus colli tendinitis, is an uncommon benign condition presenting as acute neck pain. Clinically, it can be misdiagnosed as retropharyngeal abscess, traumatic injury, or infectious spondylitis. The diagnosis is made radiographically by an amorphous calcification anterior to C1-C2 and prevertebral soft tissue swelling. We present three cases of this uncommon condition to illustrate the classic findings on plain film, CT, and MRI. Recognition of the pathognomonic imaging appearance allows for easy diagnosis preventing unnecessary tests and treatment. PMID- 16344982 TI - First jurassic choristodera from Asia. AB - Although choristoderes have a good Lower Cretaceous record in Asia, they have never previously been recorded from Jurassic deposits. Here we describe fragmentary vertebral material referable to Choristodera indet. from the Middle Jurassic Balabansai Svita of the Fergana Valley, Kyrgyzstan. This provides a significant range extension for the group in Asia and shows that choristoderes already had a Pan-Laurasian distribution in the Jurassic. PMID- 16344983 TI - Molecular and biochemical characterization of puroindoline a and b alleles in Chinese landraces and historical cultivars. AB - Kernel hardness that is conditioned by puroindoline genes has a profound effect on milling, baking and end-use quality of bread wheat. In this study, 219 landraces and 166 historical cultivars from China and 12 introduced wheats were investigated for their kernel hardness and puroindoline alleles, using molecular and biochemical markers. The results indicated that frequencies of soft, mixed and hard genotypes were 42.7, 24.3, and 33.0%, respectively, in Chinese landraces and 45.2, 13.9, and 40.9% in historical cultivars. The frequencies of PINA null, Pinb-D1b and Pinb-D1p genotypes were 43.8, 12.3, and 39.7%, respectively, in hard wheat of landraces, while 48.5, 36.8, and 14.7%, respectively, in historical hard wheats. A new Pinb-D1 allele, designated Pinb-D1t, was identified in two landraces, Guangtouxianmai and Hongmai from the Guizhou province, with the characterization of a glycine to arginine substitution at position 47 in the coding region of Pinb gene. Surprisingly, a new Pina-D1 allele, designated Pina D1m, was detected in the landrace Hongheshang, from the Jiangsu province, with the characterization of a proline to serine substitution at position 35 in the coding region of Pina gene; it was the first novel mutation found in bread wheat, resulting in a hard endosperm with PINA expression. Among the PINA null genotypes, an allele designed as Pina-D1l, was detected in five landraces with a cytosine deletion at position 265 in Pina locus; while another novel Pina-D1 allele, designed as Pina-D1n, was identified in six landraces, with the characterization of an amino acid change from tryptophan-43 to a 'stop' codon in the coding region of Pina gene. The study of puroindoline polymorphism in Chinese wheat germplasm could provide useful information for the further understanding of the molecular basis of kernel hardness in bread wheat. PMID- 16344984 TI - Trends in population parameters and best linear unbiased prediction of progeny performance in a European F(2) maize population under modified recurrent full-sib selection. AB - Recurrent selection is a cyclic breeding procedure designed to improve the mean of a population for the trait(s) under selection. Starting from an F(2) population of European flint maize (Zea mays L.) intermated for three generations, we conducted seven cycles of a modified recurrent full-sib (FS) selection scheme. The objectives of our study were to (1) monitor trends across selection cycles in the estimates of the population mean, additive and dominance variances, (2) compare predicted and realized selection responses, and (3) investigate the usefulness of best linear unbiased prediction (BLUP) of progeny performance under the recurrent FS selection scheme applied. Recurrent FS selection was conducted at three locations using a selection rate of 25% for a selection index, based on grain yield and grain moisture. Recombination was performed according to a pseudo-factorial mating scheme, where the selected FS families were divided into an upper-ranking group of parents mated to the lower ranking group. Variance components were estimated with restricted maximum likelihood. Average grain yield increased 9.1% per cycle, average grain moisture decreased 1.1% per cycle, and the selection index increased 11.2% per cycle. For the three traits we observed, no significant changes in additive and dominance variances occurred, suggesting future selection response at or near current rates of progress. Predictions of FS family performance in Cn+1 based on mean performance of parental FS families in Cn were of equal or higher precision as those based on the mean additive genetic BLUP of their parents, and corresponding correlations were of moderate size only for grain moisture. The significant increase in grain yield combined with the decrease in grain moisture suggest that the F(2) source population with use of a pseudo-factorial mating scheme is an appealing alternative to other types of source materials and random mating schemes commonly used in recurrent selection. PMID- 16344985 TI - Valproic acid selectively inhibits conversion of arachidonic acid to arachidonoyl CoA by brain microsomal long-chain fatty acyl-CoA synthetases: relevance to bipolar disorder. AB - RATIONALE: Several drugs used to treat bipolar disorder (lithium and carbamazepine), when administered chronically to rats, reduce the turnover of arachidonic acid, but not docosahexaenoic acid, in brain phospholipids by decreasing the activity of an arachidonic acid-selective phospholipase A(2). Although chronic valproic acid produces similar effects on brain arachidonic acid and docosahexaenoic acid turnover, it does not alter phospholipase A(2) activity, suggesting that it targets a different enzyme in the turnover pathway. MATERIALS AND METHODS/RESULTS: By isolating rat brain microsomal long-chain fatty acyl-CoA synthetases (Acsl), we show in vitro that valproic acid is a non-competitive inhibitor of Acsl, as it reduces the maximal velocity of the reaction without changing the affinity of the substrate for the enzyme. While valproic acid inhibited the synthesis of arachidonoyl-CoA, palmitoyl-CoA, and docosahexaenoyl CoA, the K (i )for inhibition of arachidonoyl-CoA synthesis (14.1 mM) was approximately one fifth the K (i) for inhibiting palmitoyl-CoA (85.4 mM) and docosahexaenoyl-CoA (78.2 mM) synthesis. As chronic administration of valproic acid in bipolar disorder achieves whole-brain levels of 1.0 to 1.5 mM, inhibition of arachidonoyl-CoA formation can occur at brain concentrations that are therapeutically relevant to this disease. Furthermore, brain microsomal Acsl did not produce valproyl-CoA. CONCLUSIONS: This study shows that valproic acid acts as a non-competitive inhibitor of brain microsomal Acsl, and that inhibition is substrate-selective. The study supports the hypothesis that valproic acid acts in bipolar disorder by reducing the brain arachidonic acid cascade, by inhibiting arachidonoyl-CoA formation. PMID- 16344986 TI - The effect of the adenosine A(2A) antagonist KW-6002 on motor and motivational processes in the rat. AB - RATIONALE: It is well established that humans and rats respond to an imperative stimulus more rapidly as a function of the foreperiod preceding the target, and with this decrease in mean response time, there is also an increase in anticipatory (prior to the signal) responses. These changes reflect enhanced motor readiness. Also, reaction times are quicker when the cost of reward (amount of work required) is minimum. Antagonism of the adenosine A(2A) receptor has been shown to effect motor-related processes. OBJECTIVE: This study examined the behavioural effects of systemic administration of the adenosine A(2A) antagonist KW-6002 in a cued reaction time task in the rat. The purpose of this study is to ascertain whether KW-6002 would enhance motor readiness and effect performance as a function of reward cost. METHODS: Rats were trained on a visually cued reaction time task with variable foreperiods, and the effects of three doses of KW-6002 (0.3, 1.0 and 3.0 mg/kg systemically, compared to vehicle) were examined. RESULTS: Increasing doses of KW-6002 resulted in faster reaction times and an increase in the number of anticipatory responses. KW-6002 enhanced the foreperiod effect on reaction time distributions and anticipatory responses. In addition, KW 6002 had differential effects on performance between rewarded and unrewarded trials. CONCLUSION: Antagonism of adenosine A(2A) receptors by systemic KW-6002 speeds reaction time and enhanced motor preparatory processes as well as interacting with motivational processes. PMID- 16344988 TI - A new way to generate cytolytic tumor-specific T cells: electroporation of RNA coding for a T cell receptor into T lymphocytes. AB - Effective T cell receptor (TCR) transfer until now required stable retroviral transduction. However, retroviral transduction poses the threat of irreversible genetic manipulation of autologous cells. We, therefore, used optimized RNA transfection for transient manipulation. The transfection efficiency, using EGFP RNA, was >90%. The electroporation of primary T cells, isolated from blood, with TCR-coding RNA resulted in functional cytotoxic T lymphocytes (CTLs) (>60% killing at an effector to target ratio of 20:1) with the same HLA-A2/gp100 specificity as the parental CTL clone. The TCR-transfected T cells specifically recognized peptide-pulsed T2 cells, or dendritic cells electroporated with gp100 coding RNA, in an IFNgamma-secretion assay and retained this ability, even after cryopreservation, over 3 days. Most importantly, we show here for the first time that the electroporated T cells also displayed cytotoxicity, and specifically lysed peptide-loaded T2 cells and HLA-A2+/gp100+ melanoma cells over a period of at least 72 h. Peptide-titration studies showed that the lytic efficiency of the RNA-transfected T cells was similar to that of retrovirally transduced T cells, and approximated that of the parental CTL clone. Functional TCR transfer by RNA electroporation is now possible without the disadvantages of retroviral transduction, and forms a new strategy for the immunotherapy of cancer. PMID- 16344987 TI - Ethanol-induced conditioned place preference, but not aversion, is blocked by treatment with D -penicillamine, an inactivation agent for acetaldehyde. AB - RATIONALE: There is evidence to suggest that acetaldehyde is involved in the control of ethanol-seeking behavior and reward. D -penicillamine, a thiol amino acid, is a highly selective agent for the inactivation of acetaldehyde. Previous studies from our laboratory have demonstrated that D -penicillamine prevents both behavioral stimulation induced by ethanol and acetaldehyde-produced locomotor depression in mice. OBJECTIVES: The contribution of ethanol-derived acetaldehyde to the affective effects of ethanol (preference and aversion) was assessed using an unbiased place conditioning design. METHODS: Male mice received four pairings of a distinctive floor stimulus (CS+: GRID+ or HOLE+) with injections of saline and ethanol (2 g/kg) given before (preference) or after (aversion) the 5-min exposure to the place conditioning apparatus. A different floor stimulus (CS-: GRID- or HOLE-), associated with saline-saline injections on alternate days, was presented. For a different group of animals, the pairings with the CS+ were associated with saline and ethanol injections, but on alternate days, they received D -penicillamine (50 or 75 mg/kg) and ethanol injections paired with the CS-floor stimulus. A 60-min preference test was carried out 24 h after the last conditioning trial. A similar procedure was followed to test the effect of D penicillamine on morphine (16 mg/kg) and cocaine-induced (20 mg/kg) conditioned place preference (CPP). RESULTS: CPP and conditioned place aversion (CPA) were observed for ethanol, but D -penicillamine only blocked CPP. D -penicillamine, by itself, did not produce either rewarding or aversive effects. CPP observed for morphine and cocaine was unaffected by D -penicillamine pretreatment. CONCLUSIONS: The results of the present study suggest that the selective inactivation of acetaldehyde blocked the rewarding, but not aversive, effects of ethanol and support the role of this ethanol metabolite in the affective properties of ethanol. PMID- 16344989 TI - Correlates of oxidative stress in wild kestrel nestlings (Falco tinnunculus). AB - The fitness of an organism can be affected by conditions experienced during early development. In light of the impact that oxidative stress can have on the health and ageing of a bird species, this study evaluated factors accounting for the variation in oxidative stress levels in nestlings of the Eurasian kestrel (Falco tinnunculus) by measuring the serum concentration of reactive oxygen metabolites and the serum antioxidant barrier against hypochlorite-induced oxidation. The ratio between these two variables was considered as an index of oxidative stress, with higher values meaning higher oxidative damage. Six-chick broods showed the highest level of oxidative stress, while no effect of sex was found. Age showed an inverse relationship with the oxidants and the levels of oxidative stress, with younger birds having higher levels. Hatching date, body condition, body mass and carotenoid concentration did not show any relationship with oxidants, antioxidants or degree of oxidative stress. These findings suggest that intrabrood sibling competition could play a role in determining oxidative stress, and that in carnivorous birds other antioxidant molecules could be more important than carotenoids to reduce oxidative stress. PMID- 16344990 TI - Cortisol increases the activities of intestinal apical membrane hydrolases and nutrient transporters before weaning in mink (Mustela vison). AB - Glucocorticoids from endogenous and exogenous sources accelerate maturation of brush-border membrane (BBM) hydrolases in omnivorous laboratory rodents and pigs. Less is known for carnivores, and whether the route of administration (oral or systemic) has an influence. The present study examined the influence of administering cortisol (hydrocortisone succinate, 5 mg/kg-day) to mink during postnatal week 4, just prior to weaning, on small intestine glucose and amino acid (aspartate, leucine, lysine, methionine, proline) absorption and on the activities of BBM disaccharidases and peptidases. Kits treated with cortisol were smaller (P<0.05), but had small intestines that were proportionally larger (P<0.05 for length and mass per kg body weight, but not for mucosal mass) than control kits with higher rates of absorption for most nutrients, except leucine, and increased activities of most BBM hydrolases, except lactase. As a consequence, cortisol increased hydrolytic and absorptive capacities of the entire small intestine, with the responses more pronounced when the cortisol was given orally. These findings indicate administration of cortisol stimulates growth of the developing mink small intestine, but does not accelerate the postnatal declines in nutrient transport, and may be a dam-to-kit signal that prepares suckling mink to digest and absorb the adult diet. PMID- 16344991 TI - Elevated plasma homocysteine in obese schoolchildren with early atherosclerosis. AB - Elevated plasma homocysteine is widely seen as an independent risk factor of cardiovascular disease in adults. In order to investigate the role of homocysteine in a paediatric population at risk for early atheroclerosis, we studied plasma homocysteine in obese schoolchildren and non-obese peers. Plasma homocysteine, serum vitamin B12 and serum folic acid were determined in 41 obese and 27 control subjects and related to carotid intima-media thickness and flow mediated dilatation measured on high-resolution ultrasonography. Homocysteine, vitamin B12 and folic acid were all significantly elevated in obese children. In girls, plasma homocysteine correlated significantly with body mass index (r=0.56, p=0.002), increased ICA intima-media thickness (r=0.39, p=0.035) and flow mediated dilatation (r=-0.40, p=0.031). In boys, none of these associations reached significance (all p>0.234). No independent association of homocysteine with IMT and FMD was seen after adjustment for BMI. CONCLUSION: Plasma homocysteine is elevated in obese schoolchildren with hypertension and dyslipidaemia, particularly in girls. This may indicate a high-risk constellation, so that plasma homocysteine should be monitored in these children. PMID- 16344992 TI - Uncommon synovial cysts in children. AB - Popliteal synovial cysts (Baker's cysts) are a common occurrence in children and adults. Synovial cysts in other locations and/or with atypical extension are less common and may be confounded with tumors or other medical conditions. In this article we describe the underlying disease, clinical presentation and clinical course in six children with a sudden onset of paraarticular soft tissue masses or non-specific chronic pain. Ultrasound was the initial imaging method used in all cases; this was supplemented by MRI in three patients. Four children were diagnosed to be suffering from juvenile idiopathic arthritis (JIA), one child from Lyme Arthritis, whereas in one child no underlying disease was identified. Well-demarcated hypoechogenic lesions without signs of perfusion extending from the shoulder (two patients), elbow (one patient), hip (one patient), knee (1) or ankle (one patient) far into the adjacent musculature were detected on ultrasonography. A direct connection to the joint was demonstrated in all cases. All synovial cysts in the five arthritic patients resolved rapidly with medical treatment for arthritis, whereas the cyst persisted in the non-JIA patient. CONCLUSION: Uncommon synovial cysts occur in particular as a complication of arthritis. Ultrasonography is the initial and follow-up imaging method of choice, which can be supplemented by MRI in unusual cases. Apart from treatment for arthritis, no specific therapeutic interventions were required in the present cases. PMID- 16344993 TI - Maturity-associated variation in the body size and proportions of elite female gymnasts 14-17 years of age. AB - The aim of this study was to evaluate variations in the size and body proportions of elite female gymnasts associated with individual differences in maturity status. The subjects were 150 gymnasts, 14.0-17.9 years of age, who had participated in the 24th World Championship in Artistic Gymnastics, Rotterdam. Height, body mass, sitting height and biacromial and bicristal breadths were measured. Leg length was estimated as the height minus sitting height, and the ratio of sitting height to height was calculated. Information on menarcheal status was obtained by means of a questionnaire, and hand-wrist radiographs were taken to assess skeletal maturity status. For the purpose of data analysis the gymnasts were divided into three maturity groups within each single-year chronological age (CA) group from 14 to 17 years: pre-menarcheal (n=65); post menarcheal but not skeletally mature (SA<16.0 years, n=37); post-menarcheal, skeletally mature (n=48). Differences among the groups were tested with analysis of variance and post hoc Tukey tests. The results showed that pre-menarcheal gymnasts are smaller in all dimensions compared to post-menarcheal gymnasts in all age groups but that the significance of differences varies. Post-menarcheal, skeletally mature gymnasts are heavier than pre-menarcheal gymnasts, but weight does not differ between gymnasts in the two post-menarcheal groups (with one exception, 16 years). Post-menarcheal, skeletally mature gymnasts have proportionally shorter legs than gymnasts in the other two maturity categories. Elite gymnasts of contrasting maturity status show similar trends in body size and proportions similar to those of non-athlete adolescent girls of contrasting maturity status. The results highlight the need to consider maturity-associated variation in the body dimensions of gymnasts before attributing their characteristics to the demands of regular training. PMID- 16344994 TI - Demonstration of a right inguinal hernia containing urinary bladder diverticulum on whole-body bone scan and pelvic CT. PMID- 16344995 TI - The attentional blink is not a unitary phenomenon. AB - Identification of the second of two targets is impaired if it is presented less than about 500 ms after the first. Three models of this second-target deficit, known as attentional blink (AB), were compared: resource-depletion, bottleneck, and temporary loss of control (TLC). Five experiments, in which three sequential targets were inserted in a stream of distractors, showed that identification accuracy for the leading target depended on an attentional switch whose magnitude varied with distractor-target similarity. In contrast, accuracy for the trailing target depended on similarity between the target and the trailing mask. These results strongly suggest that the AB is not a unitary phenomenon. Resource depletion was ruled out as a viable account. The effect of attentional switching was handled naturally by the TLC model, while bottleneck models offered the best account of the effect of backward masking. PMID- 16344996 TI - Study of hip joint dislocation after total hip arthroplasty. AB - The present study was undertaken to identify the factors responsible for hip joint dislocation after total hip arthroplasty, laying emphasis on analysis of the background variables of the patients. Of the 317 hips included in the study, ten (3.2%) dislocated. Only the anteversion angle of the cup differed significantly between the dislocation group and the dislocation-free group. The safe zone of the anteversion angle seems to be between 20 and 30 degrees. but it is also essential to set the antetorsion angle of the stem to match the shape of individual bones to create a more stable hip joint. This safe zone may be expanded by the additive effect of antetorsion angle of the stem. PMID- 16345000 TI - Structure-function correlation of outer membrane protein porin from Paracoccus denitrificans. AB - Porins from outer membrane of Gram-negative bacteria have a highly stable structure. Our previous studies on porin from Paracoccus denitrificans showed that the outer membrane protein porin is extremely stable toward heat, pH, and chemical denaturants. The major question we have addressed in this paper is whether the high stability of porin is a consequence of the beta-barrel structure and whether it is required for its function. To explain this we have analyzed two cases: first, we used porin wild-type and mutants and compared their structure and function; second, we compared the activity of porin preheated to different temperatures. Structural changes were monitored by infrared spectroscopy. We observed that the structural stability of porin is not equivalent to functional activity as minor alteration in the structure can result in drastic differences in the activity of porins. PMID- 16344997 TI - Giardia lamblia behavior under cytochalasins treatment. AB - Giardia lamblia, a flagellated protist, is the parasite most commonly found in the intestinal tract of humans and other mammals causing a disease known as giardiasis. This parasite presents several cytoskeletal structures whose major components are microtubules, namely: the ventral adhesive disk, eight flagella axonemes, the median body, and funis. However, the cytoskeletal filamentous structures are poorly understood, and therefore, less studied. In the present work, we used actin-interacting drugs such as cytochalasin B and D to investigate their effects on Giardia ultrastructure. Axenically grown G. lamblia trophozoites were treated with these drugs and analyzed by fluorescence microscopy and scanning and transmission electron microscopy. It was observed that trophozoites became completely misshapen, detached from the glass surface, and failed to complete cell division. The main alterations observed included: (1) disk fragmentation, (2) presence of large vacuoles, (3) alterations in flagella number and flagella internalization, (4) blocked cytokinesis but not the karyokinesis, and (5) presence of membrane undulations and blebs. These findings are discussed. PMID- 16345001 TI - Analysis of the conformational profile of trishomocubane amino acid dipeptide. AB - 4-Amino-(D3)-trishomocubane-4-carboxylic acid is a constrained alpha-amino acid residue that exhibits promising conformational characteristics, i.e., helical and beta-turns. As part of the development of conformational guidelines for the design of peptides and protein surrogates, the conformational energy calculations on trishomocubane using molecular mechanics and ab initio methods are presented. The C(alpha) carbon of trishomocubane forms part of the cyclic structure, and consequently a peptidic environment was simulated with an acetyl group on its N terminus and a methylamide group on its C-terminus. Ramachandran maps computed at the molecular mechanics level using the standard AMBER (parm94) force field libraries compared reasonably well with the corresponding maps computed at the Hartree Fock level, using the 6-31G* basis set. Trishomocubane peptide (Ac-Tris NHMe) is characterized by four low energy conformers corresponding to the C7ax, C7eq, 3(10), and alpha(L) helical structures. PMID- 16345003 TI - Studies of DNA dumbbells VIII. Melting analysis of DNA dumbbells with dinucleotide repeat stem sequences. AB - Melting curves and circular dichroism spectra were measured for a number of DNA dumbbell and linear molecules containing dinucleotide repeat sequences of different lengths. To study effects of different sequences on the melting and spectroscopic properties, six DNA dumbbells whose stems contain the central sequences (AA)(10), (AC)(10), (AG)(10), (AT)(10), (GC)(10), and (GG)(10) were prepared. These represent the minimal set of 10 possible dinucleotide repeats. To study effects of dinucleotide repeat length, dumbbells with the central sequences (AG)(n), n = 5 and 20, were prepared. Control molecules, dumbbells with a random central sequence, (RN)(n), n = 5, 10, and 20, were also prepared. The central sequence of each dumbbell was flanked on both sides by the same 12 base pairs and T(4) end-loops. Melting curves were measured by optical absorbance and differential scanning calorimetry in solvents containing 25, 55, 85, and 115 mM Na(+). CD spectra were collected from 20 to 45 degrees C and [Na(+)] from 25 to 115 mM. The spectral database did not reveal any apparent temperature dependence in the pretransition region. Analysis of the melting thermodynamics evaluated as a function of Na(+) provided a means for quantitatively estimating the counterion release with melting for the different sequences. Results show a very definite sequence dependence, indicating the salt-dependent properties of duplex DNA are also sequence dependent. Linear DNA molecules containing the (AG)(n) and (RN)(n), sequences, n = 5, 10, 20, and 30, were also prepared and studied. The linear DNA molecules had the exact sequences of the dumbbell stems. That is, the central repeat sequence in each linear duplex was flanked on both sides by the same 12-bp sequence. Melting and CD studies were also performed on the linear DNA molecules. Comparison of results obtained for the same sequences in dumbbell and linear molecular environments reveals several interesting features of the interplay between sequence-dependent structural variability, sequence length, and the unconstrained (linear) or constrained (dumbbell) molecular environments. PMID- 16345002 TI - Low versus high molecular weight poly(ethylene glycol)-induced states of stem bromelain at low pH: stabilization of molten globule and unfolded states. AB - The effect of low, medium, and high molecular weight poly(ethylene glycol) (e.g., PEG-400, -6000, and -20,000) on the structure of the acid unfolded state of unmodified stem bromelain (SB) obtained at pH 2.0 has been studied by various spectroscopic methods. The conformation of stem bromelain at pH 2.0 exhibits substantial loss of secondary structure and almost complete loss of native tertiary contacts and has been termed the acid unfolded state (A(U)). Addition of PEG-400 to A(U) led to an increase in the mean residue ellipticity (MRE) value at 222 nm, indicating formation of alpha-helical structure. On the other hand, PEG 6000 and 20,000 led to a decrease in the MRE value at 222 nm, indicating unfolding of the A(U) state. Interestingly, at 70% (w/v) PEG-400 and 40% (w/v) PEG-20,000, MRE values at 222 nm almost approach the native state at pH 7.0 and the unfolded state (6 M GnHCl) of stem bromelain, respectively. The probes for tertiary structure showed formation of nonnative tertiary contacts in the presence of 70% (w/v) PEG-400, while 40% (w/v) PEG-6000 and 20,000 were found to stabilize the unfolded state of SB. An increase in binding of 1-anilino 8 naphthalene sulfonic acid and a decrease in fractional accessibility of tryptophan residues (f(a)) compared to A(U) in the presence of 70% PEG-400 indicate that the PEG-400-induced state has a significant amount of exposed hydrophobic patches and is more compact than A(U). The results imply that the PEG 400-induced state has characteristics of molten globule, and higher molecular weight PEGs led to the unfolding of the A(U) state. PMID- 16345004 TI - The lost direction in binocular vision: the neglected signs posted by Wells, Towne, and LeConte. AB - Studies of vision have informed theories first in philosophy and then in psychology. Over the centuries, an increasing number of phenomena have been enlisted to refute or reinforce particular theories. Nowhere has this been more evident than in binocular vision. How we see a single world with two eyes is one of the oldest and most consistently studied topics in vision research. It has been discussed at least since the time of Aristotle and it has been examined experimentally since the second century, when Ptolemy defined lines of visual correspondence for the two eyes. Prior to Wheatstone's invention of the stereoscope in the 1830s, binocular vision had been studied in terms of visual directions. The stereoscope established distance (or depth) as well as direction as dimensions of binocular vision. Subsequently, depth rather than direction has been the principal concern of students of vision, and texts in English devoted to analyses of direction rather than depth have been neglected. We examine the experiments on binocular visual direction conducted by Wells before Wheatstone, and by Towne and LeConte after him, and discuss the reasons for their neglect. PMID- 16345007 TI - Walter Miles, Pop Warner, B. C. Graves, and the psychology of football. AB - In 1926-1927, a graduate student, B. C. Graves, working with Stanford University psychologist Walter Miles and legendary football coach Pop Warner, conducted an investigation of variations in signal calling as they affected the charging times of football players. The study was one of two that involved Miles and the ingenious multiple chronograph that he had invented to time the reactions of seven players simultaneously. These studies represented a brief digression in the career of Miles, who certainly was no sport psychologist. They tell of an interesting collaboration between scientist and coaches that produced one of the richest studies in sport psychology in the first half of the twentieth century. PMID- 16345008 TI - G. H. Mead in the history of sociological ideas. AB - My aim is to discuss the history of the reception of George Herbert Mead's ideas in sociology. After discussing the methodological debate between presentism and historicism, I address the interpretations of those responsible for Mead's inclusion in the sociological canon: Herbert Blumer, Jurgen Habermas, and Hans Joas. In the concluding section, I assess these reconstructions of Mead's thought and suggest an alternative more consistent with my initial methodological remarks. In particular, I advocate a reconstruction of Mead's ideas that apprehends simultaneously its evolution over time and its thematic breadth. Such a historically minded reconstruction can be not only a useful corrective to possible anachronisms incurred by contemporary social theorists, but also a fruitful resource for their theory-building endeavors. Only then can meaningful and enriching dialogue with Mead begin. PMID- 16345009 TI - Medicalizing melancholia: exploring profiles of psychiatric professionalization. AB - The nineteenth century was the site of radical changes in understanding mental illness. The professionalization of psychiatry consisted primarily of the discipline's aspiration to the status of an expert medical subspecialty. While all forms of insanity were eventually reframed in medical terms, melancholia--for moral and nosological reasons--assumed a special role that made it an ideal diagnosis for conceptual reframing. Our analysis of the journal literature of the nineteenth and early twentieth centuries in North America and Germany traces several ways in which melancholia was medicalized. As the care for the insane shifted into the professional realm of physicians and medical terminology came to replace prior descriptors of mental illness, melancholia was replaced by depression. In addition, the process of delineating affective pathology assumed a distinctly medical flavor. Finally, melancholia was firmly medicalized when its boundaries blurred with neurasthenia. Differences in how ordinary affective terms became medicalized in German and North American psychiatry illustrate the importance of local historical approaches. PMID- 16345010 TI - The influence of curcumin and manganese complex of curcumin on cadmium-induced oxidative damage and trace elements status in tissues of mice. AB - Curcumin (diferuoyl methane) from turmeric is a well-known biologically active compound. It has been shown to ameliorate oxidative stress and it is considered to be a potent cancer chemopreventive agent. In our previous study the antioxidative effects of curcumin in cadmium exposed animals were demonstrated. Also manganese exerts protective effects in experimental cadmium intoxication. The present study examined the ability of the manganese complex of curcumin (Mn curcumin) and curcumin to protect against oxidative damage and changes in trace element status in cadmium-intoxicated male mice. Curcumin or Mn-curcumin were administered at equimolar doses (0.14 mmol/kg b.w.) for 3 days, by gastric gavages, dispersed in methylcellulose. One hour after the last dose of antioxidants, cadmium chloride (33 micromol/kg) was administered subcutaneously. Both curcumin and Mn-curcumin prevented the increase of hepatic lipid peroxidation -- expressed as MDA level, induced by cadmium intoxication and attenuated the Cd-induced decrease of hepatic GSH level. No change in hepatic glutathione peroxidase or catalase activities was found in Cd-exposed mice. A decreased GSH-Px activity was measured in curcumin and Mn-curcumin alone treated mice. Neither curcumin nor Mn-curcumin treatment influenced cadmium distribution in the tissues and did not correct the changes in the balance of essential elements caused by Cd-treatment. The treatment with Mn-curcumin increased the Fe and Mn content in the kidneys of both control and Cd-treated mice and Fe and Cu content in the brain of control mice. In conclusion, regarding the antioxidative action, introducing manganese into the curcumin molecule does not potentiate the studied effects of curcumin. PMID- 16345011 TI - Analysis of corrinoids in ovine tissues. AB - Corrinoids from various ovine tissue samples (liver, blood, small intestinal fluid and faeces) were analysed using a combination of high-performance liquid chromatography (HPLC) and a radioisotope dilution assay (RIDA) to estimate the distribution of corrinoids--the cobalamins hydroxocobalamin (OH-cbl), methylcobalamin (me-cbl) and 5'-deoxyadenosylcobalamin (ado-cbl), and cobalamin analogues--in these tissues. Samples were taken from either cobalt-deficient or cobalt-replete ewes, and ruminant and pre-ruminant lambs. In liver, ado-cbl predominated, followed by analogues, OH-cbl and me-cbl. Supplementation with either cobalt (ruminant) or vitamin B12 injections (pre-ruminant) increased the amount of ado-cbl and decreased analogues. In blood, OH-cbl predominated, followed by ado-cbl, analogues and me-cbl, respectively. In small intestinal fluid, the distribution from largest to smallest percentage was analogues, ado cbl, OH-cbl and me-cbl. In faeces, analogues constituted the greatest proportion, followed by OH-cbl, ado-cbl and me-cbl, respectively. Owing to the small sample sizes only cautionary interpretations can be made. In contrast to humans, where me-cbl constitutes the highest proportion of corrinoids in plasma and ado-cbl in the liver, in sheep the amount of ado-cbl was consistently higher than me-cbl in all tissues. This may be due to the higher metabolic need of sheep for ado-cbl due to gluconeogenesis. Analogues and OH-cbl were found in each tissue, contrary to previous postulations. The much higher amount of vitamin B12 in small intestinal fluid compared with faeces indicates that a large proportion of the vitamin is absorbed by the gastro-intestinal tract. PMID- 16345012 TI - An HPLC/MS method for identifying major constituents in the hypocholesterolemic extracts of Chinese medicine formula 'Xue-Fu-Zhu-Yu decoction'. AB - In this paper, HPLC-ESI-MS technique was used to analyze the hypocholesterolemic extracts of 'Xue-Fu-Zhu-Yu decoction', a traditional Chinese medicine consisting of six crude drugs (i.e. Paeonia lactiflora, Ligusticum chuanxiong, Citrus aurantium, Carthamus tinctorius, Prunus persica and Bupleurum falcatum). A total of 17 compounds were identified and their plant derivations were also distinguished. Nine compounds among them were unambiguously determined as oxypaeoniflorin, amygdalin, albiflorin, paeoniflorin, ferulic acid, naringin, hesperidin, senkyunolide I and neohesperidin by comparing the retention times (tR), UV spectra and m/z values with those of the reference compounds. The other eight compounds were tentatively identified as prunasin, 6-hydroxy-kaempferol 3,6 diglucoside, 6-hydroxykaempferol 3-rutinoside-6-glucoside, galloylpaeoniflorin, 6 hydroxy-kaempferol 3-glucoside, anhydrosafflor yellow B, narirutin and kaempferol 3-rutinoside by MS2 spectra and the comparison of their UV spectra and MS spectra with literature data. PMID- 16345013 TI - Identification of biologically active triterpenes and sterols present in hexane extracts from Miconia species using high-resolution gas chromatography. AB - In this work we describe the identification of the biologically active triterpenes and sterols present in the hexane extracts of six species of Miconia using gas chromatography. The main compounds present in these extracts are beta amyrin, alpha-amyrin, and beta-sitosterol. The technique employed herein is shown to be a valuable and rapid tool for determining biologically active triterpenes and sterols present in non-polar extracts. PMID- 16345016 TI - Rater bias in a blinded randomized placebo-controlled psychiatry trial. AB - Rater bias occurs when rater knowledge of treatment assignment modifies the outcome assessment. Raters may be unconsciously influenced by inclinations for or against a particular treatment and consequently may give a more or less generous assessment depending upon these biases. Blinding of raters by keeping raters unaware of treatment assignment is one way to limit bias influencing assessment due to knowledge of treatment assignment. Unblinding may be particularly problematic in efficacy studies comparing placebo to drugs and/or non-drug psychotherapy treatments where subjects may reveal drug side-effects or mention their therapist by name, thus unblinding their treatment assignment. We present a new instrumental variable statistical approach for assessing the association between success in blinding and its impact on efficacy estimates of active drug and/or cognitive behavioural psychotherapy versus placebo in the multicentre comparative treatment study of panic disorder. Despite the uncertainty involved in assessing bias that may be unobserved and unconscious, we will show how to derive a bound for the impact of rater bias. PMID- 16345017 TI - Modelling SARS data using threshold geometric process. AB - During the outbreak of an epidemic disease, for example, the severe acute respiratory syndrome (SARS), the number of daily infected cases often exhibit multiple trends: monotone increasing during the growing stage, stationary during the stabilized stage and then decreasing during the declining stage. Lam first proposed modelling a monotone trend by a geometric process (GP) [X(i), i=1,2,...] directly such that [a(i-1)X(i), i=1,2,...] forms a renewal process for some ratio a>0 which measures the direction and strength of the trend. Parameters can be conveniently estimated using the LSE methods. Previous GP models limit to data with only a single trend. For data with multiple trends, we propose a moving window technique to locate the turning point(s). The threshold GP model is fitted to the SARS data from four regions in 2003. PMID- 16345018 TI - Pseudo-likelihood methods for longitudinal binary data with non-ignorable missing responses and covariates. AB - In this paper we consider longitudinal studies in which the outcome to be measured over time is binary, and the covariates of interest are categorical. In longitudinal studies it is common for the outcomes and any time-varying covariates to be missing due to missed study visits, resulting in non-monotone patterns of missingness. Moreover, the reasons for missed visits may be related to the specific values of the response and/or covariates that should have been obtained, i.e. missingness is non-ignorable. With non-monotone non-ignorable missing response and covariate data, a full likelihood approach is quite complicated, and maximum likelihood estimation can be computationally prohibitive when there are many occasions of follow-up. Furthermore, the full likelihood must be correctly specified to obtain consistent parameter estimates. We propose a pseudo-likelihood method for jointly estimating the covariate effects on the marginal probabilities of the outcomes and the parameters of the missing data mechanism. The pseudo-likelihood requires specification of the marginal distributions of the missingness indicator, outcome, and possibly missing covariates at each occasions, but avoids making assumptions about the joint distribution of the data at two or more occasions. Thus, the proposed method can be considered semi-parametric. The proposed method is an extension of the pseudo likelihood approach in Troxel et al. to handle binary responses and possibly missing time-varying covariates. The method is illustrated using data from the Six Cities study, a longitudinal study of the health effects of air pollution. PMID- 16345019 TI - Operating characteristics of sample size re-estimation with futility stopping based on conditional power. AB - Various methods have been described for re-estimating the final sample size in a clinical trial based on an interim assessment of the treatment effect. Many re weight the observations after re-sizing so as to control the pursuant inflation in the type I error probability alpha. Lan and Trost (Estimation of parameters and sample size re-estimation. Proceedings of the American Statistical Association Biopharmaceutical Section 1997; 48-51) proposed a simple procedure based on conditional power calculated under the current trend in the data (CPT). The study is terminated for futility if CPT < or = CL, continued unchanged if CPT > or = CU, or re-sized by a factor m to yield CPT = CU if CL < CPT < CU, where CL and CU are pre-specified probability levels. The overall level alpha can be preserved since the reduction due to stopping for futility can balance the inflation due to sample size re-estimation, thus permitting any form of final analysis with no re-weighting. Herein the statistical properties of this approach are described including an evaluation of the probabilities of stopping for futility or re-sizing, the distribution of the re-sizing factor m, and the unconditional type I and II error probabilities alpha and beta. Since futility stopping does not allow a type I error but commits a type II error, then as the probability of stopping for futility increases, alpha decreases and beta increases. An iterative procedure is described for choice of the critical test value and the futility stopping boundary so as to ensure that specified alpha and beta are obtained. However, inflation in beta is controlled by reducing the probability of futility stopping, that in turn dramatically increases the possible re-sizing factor m. The procedure is also generalized to limit the maximum sample size inflation factor, such as at m max = 4. However, doing so then allows for a non-trivial fraction of studies to be re-sized at this level that still have low conditional power. These properties also apply to other methods for sample size re-estimation with a provision for stopping for futility. Sample size re-estimation procedures should be used with caution and the impact on the overall type II error probability should be assessed. PMID- 16345020 TI - The analysis of contingency tables with ordinal data: an application to monitoring antibiotic resistance. AB - Rationalization of antibiotic therapy in the management of infectious diseases is helped by a knowledge of the patterns of sensitivity and resistance of bacteria to antibiotics and their possible changes both in time and from one hospital unit to another. In this paper we present the results regarding the sensitivities of several groups of bacteria and different Units of the S.Orsola-Malpighi Hospital of Bologna in the period 1995-1997. We apply recent methods of analysis of ordinal contingency tables that rely on stochastic ordering of the rows to test the assumption that a decrease (or increase) in sensitivity of bacteria to specific antibiotics has taken place against the alternative that no such thing has happened. In most cases the results seem to indicate an increase in sensitivity rather than what was expected, namely the opposite. PMID- 16345021 TI - Cumulative effects of air pollution on public health. AB - Cumulative effect is an important way through which the pollutants affect public health. However, few existing dynamical models are well enough understood and documented to detect or quantify the cumulative effects and to answer pertinent questions posed by the World Health Organization (WHO): 'Is there a threshold below which no effects of the pollutants on health are expected to occur in all people?' and 'What averaging period (time pattern) is the most relevant from the point of view of health?'. Using a new semi-parametric time series modelling approach, which incorporates non-linearity and latent cumulative variables, we show that the cumulative effects on health due to continual exposure to environmental pollutants can be very serious even at levels below the national ambient air quality standards of America (NAAQS). The situation is especially worrying for chronic sufferers. Our study suggests that different pollutants may require different cumulative periods (on average) to impact on health but they share a similar functional form in respect of their impact. We also suggest some possible revision of the ambient air quality standards. PMID- 16345022 TI - Mixture model analysis for establishing a diagnostic cut-off point for pertussis antibody levels. AB - Previous studies of pertussis (whooping cough) that have derived diagnostic cut off points for pertussis antibody levels have assumed a single distribution for antibody levels and have used small sample sizes. In a recent study of 5409 serum samples from the Third National Health and Nutrition Examination Survey (NHANES III), a finite mixture model was developed to examine the distribution of immunoglobulin G (IgG) antibody levels against pertussis toxin (PT), an antigen specific to the Bordetella pertussis bacterium. The mixture model identified three component populations with antibody levels greater than the quantitative assay's lower limit of quantitation (LLQ) and included a point distribution located at or below the LLQ to account for the excess number of antibody values that fell below the LLQ. The mixture model analysis accounted for the NHANES III design. A cut-off point for anti-PT IgG levels was chosen to have a 99 per cent model specificity based on the two overlapping normal distributions assumed for the two component populations with the highest antibody levels. This cut-off point may have a higher diagnostic sensitivity for acute B. pertussis infection than other cut-off points derived by assuming a single distribution for antibody levels. PMID- 16345024 TI - Shared random effects analysis of multi-state Markov models: application to a longitudinal study of transitions to dementia. AB - Multi-state models are appealing tools for analysing data about the progression of a disease over time. In this paper, we consider a multi-state Markov chain with two competing absorbing states: dementia and death and three transient non demented states: cognitively normal, amnestic mild cognitive impairment (amnestic MCI), and non-amnestic mild cognitive impairment (non-amnestic MCI). The likelihood function for the data is derived and estimates for the effects of the covariates on transitions are determined when the process can be viewed as a polytomous logistic regression model with shared random effects. The presence of a shared random effect not only complicates the formulation of the likelihood but also its evaluation and maximization. Three approaches for maximizing the likelihood are compared using a simulation study; the first method is based on the Gauss-quadrature technique, the second method is based on importance sampling ideas, and the third method is based on an expansion by Taylor series. The best approach is illustrated using a longitudinal study on a cohort of cognitively normal subjects, followed annually for conversion to mild cognitive impairment (MCI) and/or dementia, conducted at the Sanders Brown Center on Aging at the University of Kentucky. PMID- 16345025 TI - Modelling menstrual status during and after adjuvant treatment for breast cancer. AB - Failure time data may consist of the observation of an event whose cause is unknown due to the censoring or lack of a second event that could identify the cause of the first event. Standard competing-risks methodology does not apply to this setting because the cause of the event is not always identifiable. Moreover, one cannot assume that the entire population will eventually experience the event of interest, and the observation is potentially censored for all patients. The model that we describe in this article is motivated by a breast cancer clinical trial conducted by the International Breast Cancer Study Group (IBCSG). Because some breast cancer adjuvant treatments for premenopausal patients who have undergone surgery cause the interruption of menses, or amenorrhoea, it is of interest to describe the process by which menses discontinue and resume after treatment is completed. The process is complicated by the fact that natural menopause also occurs in the patient population, and that treatment-induced amenorrhoea is not distinguishable from menopause unless menses are observed to resume after treatment completion. We discuss a parametric model for the time to amenorrhoea and for the time to the recovery of menses, also accounting for the presence of censoring and for the possibility that treatment causes an anticipation of natural menopause. PMID- 16345026 TI - Repeated events survival models: the conditional frailty model. AB - Repeated events processes are ubiquitous across a great range of important health, medical, and public policy applications, but models for these processes have serious limitations. Alternative estimators often produce different inferences concerning treatment effects due to bias and inefficiency. We recommend a robust strategy for the estimation of effects in medical treatments, social conditions, individual behaviours, and public policy programs in repeated events survival models under three common conditions: heterogeneity across individuals, dependence across the number of events, and both heterogeneity and event dependence. We compare several models for analysing recurrent event data that exhibit both heterogeneity and event dependence. The conditional frailty model best accounts for the various conditions of heterogeneity and event dependence by using a frailty term, stratification, and gap time formulation of the risk set. We examine the performance of recurrent event models that are commonly used in applied work using Monte Carlo simulations, and apply the findings to data on chronic granulomatous disease and cystic fibrosis. PMID- 16345027 TI - A simple and general change-point identifier. AB - In a not-necessarily-stationary time-series, a Moving F statistic can identify points of change in the nature of the series model (the estimate of the underlying data-generating process), in its parameters, in residual variability, or in any combination of these. In addition, it can uncover changes masked by a reduction in residual variability. Patterns in the forces giving rise to the data may often be perceived. To form the Moving F, a theory of the process or a regression method on a baseline sample estimates the series model and the residual mean square about it is calculated. This series model is extended past the baseline with residuals assumed to be normally distributed. The Moving F is calculated as the moving average of squared deviations about the series model in ratio to the baseline mean square. The Moving F crossing the critical F identifies a change in the series model, i.e. signals its presence and location. In our experience, this Moving F method is easier to use than other commonly employed change-point identifiers (CUSUM, EWMA, data-based bandwidth selection, MCMC) and has been found to work in several situations where some other identifiers fail. (MCMC is more general, but requires advanced statistical ability.) Examples given are monitored prostate specific antigen in a post treatment prostate cancer patient and detection of Harold Shipman's medical murders. Moving F is 'simple and general' in the sense of both simultaneously; we have not found another relatively simple method to be as general. PMID- 16345028 TI - Growth and cognitive function of Indonesian children: Zero-inflated proportion models. AB - This is a study of the cognitive function of Indonesian children. The cognitive function assessment consisted of 17 items and the number of correct answers a child could score therefore ranged between 0 and 17. In this upper bounded situation, a zero-inflated binomial model and a zero-inflated beta-binomial model were considered. The purpose of the analysis was to examine whether the growth status in infancy and at the age of 7 years was related to the cognitive function of the children. The four regression models proposed by Lucas, Fewtrell and Cole were fitted, i.e. an 'early model' relating early body size to the outcome, a 'later model' relating later body size to the outcome, a 'combined model' including both early and later anthropometry measures, and an 'interaction model' further including an interaction term calculated as the product of early and later body size. It was found that social variables predicted the probability of zero-inflation, while weight-for-age at 7 years predicted the proportion of correct answers. The results do not support the existence of a critical window of cognitive development in infancy. Rather they suggest that it is the lack of catch-up growth after poor growth in infancy that is hazardous, and that childhood weight gain is influential regardless of weight in infancy. PMID- 16345029 TI - Measuring and estimating diagnostic accuracy when there are three ordinal diagnostic groups. AB - This article studies the problem of measuring and estimating the diagnostic accuracy when there are three ordinal diagnostic groups. We use a receiver operating characteristic (ROC) surface to describe the probabilities of correct classifications into three diagnostic groups based on various sets of diagnostic thresholds of a test and propose to use the entire and the partial volume under the surface to measure the diagnostic accuracy. Mathematical properties and probabilistic interpretations of the proposed measure of diagnostic accuracy are discussed. Under the assumption of normal distributions of the diagnostic test from three diagnostic groups, we present the maximum likelihood estimate to the volume under the ROC surface and give the asymptotic variance to the estimate. We further propose several asymptotic confidence interval estimates to the volume under the ROC surface. The performance of these confidence interval estimates is evaluated in terms of attaining the nominal coverage probability based on a simulation study. In addition, we develop a method of sample size determination to achieve an adequate accuracy of the confidence interval estimate. Finally, we demonstrate the proposed methodology by applying it to the clinical diagnosis of early stage Alzheimer's disease based on the neuropsychological database of the Washington University Alzheimer's Disease Research Center. PMID- 16345030 TI - cAMP cascade (PKA, Epac, adenylyl cyclase, Gi, and phosphodiesterases) regulates myelin phagocytosis mediated by complement receptor-3 and scavenger receptor AI/II in microglia and macrophages. AB - The removal by phagocytosis of degenerated myelin is central for repair in Wallerian degeneration that follows traumatic injury to axons and in autoimmune demyelinating diseases (e.g., multiple sclerosis). We tested for roles played by the cAMP cascade in the regulation of myelin phagocytosis mediated by complement receptor-3 (CR3/MAC-1) and scavenger receptor-AI/II (SRAI/II) separately and combined in mouse microglia and macrophages. Components of the cAMP cascade tested are cAMP, adenylyl cyclase (AC), Gi, protein kinase A (PKA), exchange protein directly activated by cAMP (Epac), and phosphodiesterases (PDE). PKA inhibitors H-89 and PKI(14-22) amide inhibited phagocytosis at normal operating cAMP levels (i.e., those occurring in the absence of reagents that alter cAMP levels), suggesting activation of phagocytosis through PKA at normal cAMP levels. Phagocytosis was inhibited by reagents that elevate endogenous cAMP levels to above normal: Gi-inhibitor Pertussis toxin (PTX), AC activator Forskolin, and PDE inhibitors IBMX and Rolipram. Phagocytosis was inhibited also by cAMP analogues whose addition mimics abnormal elevations in endogenous cAMP levels: nonselective 8-bromo-cAMP, PKA-specific 6-Benz-cAMP, and Epac-specific 8-CPT-2'-O-Me-cAMP, suggesting that abnormal high cAMP levels inhibit phagocytosis through PKA and Epac. Altogether, observations suggest a dual role for cAMP and PKA in phagocytosis: activation at normal cAMP levels and inhibition at higher. Furthermore, a balance between Gi-controlled cAMP production by AC and cAMP degradation by PDE maintains normal operating cAMP levels that enable efficient phagocytosis. PMID- 16345031 TI - Proteomic evaluation reveals that olfactory ensheathing cells but not Schwann cells express calponin. AB - Human clinical trials have begun worldwide that use olfactory ensheathing cells (OECs) to ameliorate the functional deficits following spinal cord injury. These trials have been initiated largely because numerous studies have reported that OECs transform into Schwann Cell (SC)-like cells that myelinate axons and support new growth in adult rats with spinal injury. This phenomenon is remarkable because OECs do not myelinate olfactory axons in their native environment. Furthermore, these myelinating OECs are morphologically identical to SCs, which can invade the spinal cord after injury. One factor that has contributed to a possible confusion in the identification of these cells is the lack of phenotypic markers to distinguish unequivocally between OECs and SCs. Such markers are required to first assess the degree of SC contamination in OEC cultures before intraspinal implantation, and then to accurately identify grafted OECs and invading SCs in the injured spinal cord. Using two-dimensional gel electrophoresis, we have identified calponin, an actin binding protein, as the first definitive phenotypic marker that distinguishes between OECs and SCs in vitro and in vivo. We have also provided ultrastructural evidence that calponin immunopositive OECs do not transform into myelinating SC-like cells after intraspinal implantation. Rather, the grafted OECs retain their morphological and neurochemical features. These data yield new insight into the phenotypic characteristics of OECs, which together with invading SCs can enhance regeneration of the injured spinal cord. PMID- 16345032 TI - Survival of neural precursor cells in growth factor-poor environment: implications for transplantation in chronic disease. AB - A key issue for therapeutic neural stem cell transplantation in chronic diseases is the long-term survival of transplanted cells in the brain. The normal adult central nervous system does not support the survival of transplanted cells. Presumably, the limited availability of trophic factors maintains the survival of resident cells but is insufficient for supporting the survival of transplanted cells. Specifically, in multiple sclerosis, a chronic relapsing disease, it would be necessary to maintain long-term survival of transplanted cells through phases of relapses and remissions. It may be beneficial to transplant cells as early as possible, in a form that will keep their survival independent of tissue support and ready for immediate mobilization upon tissue demand during disease relapse. In the present study, we examined whether, in the form of neurospheres, multipotential neural precursor cells (NPCs) survive in a growth factor-poor environment while maintaining their potential to respond to environmental cues. We found that after removal of growth factors from the culture medium of neurospheres in vitro, NPC proliferation decreased significantly, but most cells survived for a prolonged time and maintained their stem cell characteristics. After re-exposure to growth factors, neurosphere cells resumed proliferation and could differentiate along neural lineages. Furthermore, neurospheres, but not single NPCs, that were transplanted into the brain ventricles of intact animals survived within the ventricles for at least a month and responded to induction of experimental autoimmune encephalomyelitis and brain inflammation by extensive migration into the brain white matter and differentiated into glial lineage cells. PMID- 16345033 TI - ROC analysis for markers with mass at zero. AB - The receiver operating characteristic (ROC) curve and in particular the area under the curve (AUC) is commonly used to examine the discriminatory ability of diagnostic markers. Certain markers while basically continuous and non-negative have a positive probability mass (spike) at the value zero. We discuss a flexible modelling approach to such data and contrast it with the standard non-parametric approach. We show how the modelling approach can be extended to take account of the effect of explanatory variables. We motivate this problem and illustrate the modelling approach using data on the coronary calcium score, measured by electron beam tomography, which is a marker for atherosclerosis. PMID- 16345034 TI - A mixed-effects model for cognitive decline with non-monotone non-response from a two-phase longitudinal study of dementia. AB - Most longitudinal studies of elderly are characterized by substantial drop-out due to death and many other factors beyond the control of the investigators. In a two-phase longitudinal study of dementia, subjects with cognitive impairment skip the first phase survey in the next follow-up, leading to intermittent missing variables measured in that phase. In the context of analysing pre-dementia cognitive decline in an elderly population, both of the two causes of non response can potentially be informative in the sense that the missingness is dependent on the unobserved outcome. To take these factors into account, mixed effects models are constructed to allow the outcome and the multiple causes of missing values to share the same 'random parameter' or random effect. The crucial assumption of our model is that the random effects of the model for the outcome and that of the model for the missing-data indicators are linked in a deterministic manner. It can be thought of as an approximation of a more general and realistic situation, in which the two models have distinct, yet dependent, random effects. We conduct a simulation study to investigate possible deviations of the estimates under such a scenario. A second simulation illustrates the magnitude of the bias in estimating the difference of decline rate between two groups when the random effects are linked in different manners for the two groups. PMID- 16345035 TI - Bias reduction in effectiveness analyses of longitudinal ordinal doses with a mixed-effects propensity adjustment. AB - A mixed-effects propensity adjustment is described that can reduce bias in longitudinal studies involving non-equivalent comparison groups. There are two stages in this data analytic strategy. First, a model of propensity for treatment intensity examines variables that distinguish among subjects who receive various ordered doses of treatment across time using mixed-effects ordinal logistic regression. Second, the effectiveness model examines multiple times until recurrence to compare the ordered doses using a mixed-effects grouped-time survival model. Effectiveness analyses are initially stratified by propensity quintile. Then the quintile-specific results are pooled, assuming that there is not a propensity x treatment interaction. A Monte Carlo simulation study compares bias reduction in fully specified propensity model relative to misspecified models. In addition, type I error rate and statistical power are examined. The approach is illustrated by applying it to a longitudinal, observational study of maintenance treatment of major depression. PMID- 16345036 TI - Gender-based differences of percutaneous coronary intervention in the drug eluting stent era. AB - OBJECTIVES: The purpose of this study is to provide insights into percutaneous coronary intervention (PCI) performed in women in the United States by evaluating gender-based PCI-practice patterns and outcomes. BACKGROUND: Limited "real world" contemporary data exist on how the introduction of DES has impacted PCI in women. METHODS AND RESULTS: Patients (359 women, 807 men) with de novo coronary artery disease having PCI (1,166) were evaluated during the first year, since the introduction of DES in the United States market (May 1, 2003 to April 30, 2004). Women were more likely to be older, hypertensive, obese, diabetic, and have heart failure. Men were more likely to be smokers and have more vessels with obstructive coronary artery disease. PCI procedural success rates, number of vessels attempted, percentage DES utilization, and in-hospital major adverse cardiac events (MACE; death, new myocardial infarction, urgent revascularization) were similar for both genders. However, women had significantly higher unadjusted mortality (3.9% versus 1.6%, P = 0.01), cumulative vascular complications (12.0% versus 4.2%, P < 0.0001), and renal failure (2.5% versus 0.7%, P = 0.01). After adjustment for confounding variables, mortality was similar between genders, but a significant association with vascular complications and trend toward higher rates of renal failure persisted in women. CONCLUSIONS: In this study of the modern era of PCI with DES utilization, in-hospital MACE is similar between men and women. However, the differences in baseline comorbidities and the proclivity for vascular and renal complications highlight the need for further investigation and improvements to optimize outcomes of PCI in women. PMID- 16345037 TI - Investigating brain metabolism at high fields using localized 13C NMR spectroscopy without 1H decoupling. AB - Most in vivo 13C NMR spectroscopy studies in the brain have been performed using 1H decoupling during acquisition. Decoupling imposes significant constraints on the experimental setup (particularly for human studies at high magnetic field) in order to stay within safety limits for power deposition. We show here that incorporation of the 13C label from 13C-labeled glucose into brain amino acids can be monitored accurately using localized 13C NMR spectroscopy without the application of 1H decoupling. Using LCModel quantification with prior knowledge of one-bond and multiple-bond J(CH) coupling constants, the uncertainty on metabolites concentrations was only 35% to 91% higher (depending on the carbon resonance of interest) in undecoupled spectra compared to decoupled spectra in the rat brain at 9.4 Tesla. Although less sensitive, 13C NMR without decoupling dramatically reduces experimental constraints on coil setup and pulse sequence design required to keep power deposition within safety guidelines. This opens the prospect of safely measuring 13C NMR spectra in humans at varied brain locations (not only the occipital lobe) and at very high magnetic fields above 4 Tesla. PMID- 16345038 TI - A modified test for small-study effects in meta-analyses of controlled trials with binary endpoints. AB - Publication bias and related bias in meta-analysis is often examined by visually checking for asymmetry in funnel plots of treatment effect against its standard error. Formal statistical tests of funnel plot asymmetry have been proposed, but when applied to binary outcome data these can give false-positive rates that are higher than the nominal level in some situations (large treatment effects, or few events per trial, or all trials of similar sizes). We develop a modified linear regression test for funnel plot asymmetry based on the efficient score and its variance, Fisher's information. The performance of this test is compared to the other proposed tests in simulation analyses based on the characteristics of published controlled trials. When there is little or no between-trial heterogeneity, this modified test has a false-positive rate close to the nominal level while maintaining similar power to the original linear regression test ('Egger' test). When the degree of between-trial heterogeneity is large, none of the tests that have been proposed has uniformly good properties. PMID- 16345039 TI - A Bayesian stopping rule for a single arm study: With a case study of stem cell transplantation. AB - Continuous monitoring of treatment failures is an important issue in clinical studies of a single experimental treatment for high risk therapy such as hematopoietic stem cell transplantation. The sequential probability ratio test (SPRT) of Wald in 1947 and various alternative stopping rules have been proposed for sequential monitoring of adverse events. It is natural to use prior information to improve stopping rules and statistical analysis. A Bayesian stopping rule is developed and applied to an example of an umbilical cord blood transplant study performed at the University of Minnesota. Two strata, based on the number of nucleated cells per kg recipient body weight (the 'dose') are monitored separately and different rules are constructed for each stratum using different prior distributions. It is believed that patients in the lower dose group have a greater chance of graft failure than those in the higher dose group. A program, written in R, is also presented for calculating the stopping rule using the prior beliefs. The program is an improvement upon existing programs and it can be used for larger studies. PMID- 16345040 TI - High-resolution FMRI at 1.5T using balanced SSFP. AB - The resolution in conventional BOLD FMRI is considerably lower than can be achieved with other MRI methods, and is insufficient for many important applications. One major difficulty in robustly improving spatial resolution is the poor image quality in BOLD FMRI, which suffers from distortions, blurring, and signal dropout. This work considers the potential for increased resolution with a new FMRI method based on balanced SSFP. This method establishes a blood oxygenation sensitive steady-state (BOSS) signal, in which the frequency sensitivity of balanced SSFP is used to detect the frequency shift of deoxyhemoglobin. BOSS FMRI is highly SNR efficient and does not suffer from image distortions or signal dropout, making this method an excellent candidate for high resolution FMRI. This study presents the first demonstration of high-resolution BOSS FMRI, using an efficient 3D stack-of-segmented EPI readout and combined acquisition at multiple center frequencies. BOSS FMRI is shown to enable high resolution FMRI data (1 x 1 x 2 mm(3)) in both visual and motor systems using standard hardware at 1.5 T. Currently, the major limitation of BOSS FMRI is its sensitivity to temporal and spatial field drift. PMID- 16345041 TI - Non-parametric assessment of non-inferiority with censored data. AB - We suggest non-parametric tests for showing non-inferiority of a new treatment compared to a standard therapy when data are censored. To this end the difference and the odds ratio curves of the entire survivor functions over a certain time period are considered. Two asymptotic approaches for solving these testing problems are investigated, which are based on bootstrap approximations. The performance of the test procedures is investigated in a simulation study, and some guidance on which test to use in specific situations is derived. The proposed methods are applied to a trial in which two thrombolytic agents for the treatment on acute myocardial infarction were compared, and to a study on irradiation therapies for advanced non-small-cell lung cancer. Non-inferiority over a large time period of the study can be shown in both cases. PMID- 16345042 TI - On non-inferiority margin and statistical tests in active control trials. AB - The problem of selecting a non-inferiority margin and the corresponding statistical test for non-inferiority in active control trials is considered. For selection of non-inferiority margin, the guideline by the International Conference on Harmonization (ICH) recommends that the non-inferiority margin should be chosen in such a way that if the non-inferiority of the test therapy to the active control agent is claimed, the test therapy is not only non-inferior to the active control agent, but also superior to the placebo. Furthermore, variability should be taken into account. Along this line, a method for selecting non-inferiority margins with some statistical justification is proposed. Statistical tests for non-inferiority designed in the situation where the non inferiority margin is an unknown parameter are derived. An example concerning a cancer trail for testing non-inferiority with the primary study endpoint of the time to disease progression is presented to illustrate the proposed method. PMID- 16345043 TI - Longitudinal and repeated cross-sectional cluster-randomization designs using mixed effects regression for binary outcomes: bias and coverage of frequentist and Bayesian methods. AB - As medical applications for cluster randomization designs become more common, investigators look for guidance on optimal methods for estimating the effect of group-based interventions over time. This study examines two distinct cluster randomization designs: (1) the repeated cross-sectional design in which centres are followed over time but patients change, and (2) the longitudinal design in which individual patients are followed over time within treatment clusters. Simulations of each study design stipulated a multiplicative treatment effect (on the log odds scale), between 5 and 15 clusters in each of two treatment arms, and followed over two time periods. Estimation options included linear mixed effects models using restricted maximum likelihood (REML), generalized estimating equations (GEE), mixed effects logistic regression using both penalized quasi likelihood (PQL) and numerical integration, and Bayesian Monte Carlo analysis. For the repeated cross-sectional designs, most methods performed well in terms of bias and coverage when clusters were numerous (30) and variability across clusters of baseline risk and treatment effect was modest. With few clusters (two groups of five) and higher variability, only the Bayesian methods maintained coverage. In the longitudinal designs, the common methods of REML, GEE, or PQL performed poorly when compared to numerical integration, while Bayesian methods demonstrated less bias and better coverage for estimates of both log odds ratios and risk differences. The performance of common statistical tools for the analysis of cluster randomization designs depends heavily on the precise design, the number of clusters, and the variability of baseline outcomes and treatment effects across centres. PMID- 16345046 TI - Application of reliability coefficients in cDNA microarray data analysis. AB - Gene expression microarray technology has been widely used in areas such as human cancer research to identify molecular characteristics of sample specimens. The microarray study, however, is a very complicated procedure which involves numerous sources of variability that may be either systematic or random. Systematic variation is often eliminated by applying normalization procedures, but at present there are no standard criteria available to evaluate the performance of a particular normalization approach. In this paper, we propose a reliability-type coefficient as a criterion to assess the effectiveness of normalization procedures in eliminating systematic variation. Simulation studies show that this criterion performs reasonably well in a range of settings. The proposed method is illustrated using a subset of an ongoing microarray study of soft-tissue sarcoma. PMID- 16345047 TI - Survival analysis using auxiliary variables via non-parametric multiple imputation. AB - We develop an approach, based on multiple imputation, that estimates the marginal survival distribution in survival analysis using auxiliary variables to recover information for censored observations. To conduct the imputation, we use two working survival models to define a nearest neighbour imputing risk set. One model is for the event times and the other for the censoring times. Based on the imputing risk set, two non-parametric multiple imputation methods are considered: risk set imputation, and Kaplan-Meier imputation. For both methods a future event or censoring time is imputed for each censored observation. With a categorical auxiliary variable, we show that with a large number of imputes the estimates from the Kaplan-Meier imputation method correspond to the weighted Kaplan-Meier estimator. We also show that the Kaplan-Meier imputation method is robust to mis specification of either one of the two working models. In a simulation study with time independent and time-dependent auxiliary variables, we compare the multiple imputation approaches with an inverse probability of censoring weighted method. We show that all approaches can reduce bias due to dependent censoring and improve the efficiency. We apply the approaches to AIDS clinical trial data comparing ZDV and placebo, in which CD4 count is the time-dependent auxiliary variable. PMID- 16345048 TI - A Bayesian determination of threshold for identifying differentially expressed genes in microarray experiments. AB - The original definitions of false discovery rate (FDR) and false non-discovery rate (FNR) can be understood as the frequentist risks of false rejections and false non-rejections, respectively, conditional on the unknown parameter, while the Bayesian posterior FDR and posterior FNR are conditioned on the data. From a Bayesian point of view, it seems natural to take into account the uncertainties in both the parameter and the data. In this spirit, we propose averaging out the frequentist risks of false rejections and false non-rejections with respect to some prior distribution of the parameters to obtain the average FDR (AFDR) and average FNR (AFNR), respectively. A linear combination of the AFDR and AFNR, called the average Bayes error rate (ABER), is considered as an overall risk. Some useful formulas for the AFDR, AFNR and ABER are developed for normal samples with hierarchical mixture priors. The idea of finding threshold values by minimizing the ABER or controlling the AFDR is illustrated using a gene expression data set. Simulation studies show that the proposed approaches are more powerful and robust than the widely used FDR method. PMID- 16345049 TI - Magnetic resonance assessment of myocardial perfusion via catheter-based ventricle-coronary vein bypass in porcine myocardial infarction model. AB - OBJECTIVE: The goal of this study was to investigate the efficacy of VPASS with physiological measurements, magnetic resonance imaging (MRI), and histology in a porcine model of myocardial infarction. BACKGROUND: A catheter-based ventricle-to coronary vein bypass (VPASS) has been proposed as a potential treatment strategy for refractory coronary artery disease patients. METHODS: In an acute setting, the VPASS implant was deployed percutaneously in three swine. The partial pressure of oxygen (PO(2)) in the anterior interventricular vein (AIV) and left ventricle (LV) were measured before and after VPASS implant with various combinations of balloon occlusion in the AIV and left anterior descending artery (LAD). In a separate chronic study, the VPASS procedure was completed on three swine with a mid-LAD occlusion. Thirty days post-VPASS procedure, angiography, contrast-enhanced MRI, and histology were performed to assess myocardial viability. Perfusion was analyzed using the average percent signal intensity change (APSIC) in the anterior walls (AW) and inferior walls (IW). RESULTS: The VPASS implant was performed without complication. Post-VPASS implantation, the distal AIV PO(2) increased up to the LV PO(2) level during simultaneous AIV and LAD blockage (432 +/- 24 mmHg). At day 30, quantitative perfusion analysis demonstrated no difference in APSIC between AW and IW (125 +/- 26% vs. 137 +/- 38%, P = 0.46). Delayed enhancement and histology showed focal subendomyocardial infarction. CONCLUSIONS: VPASS implant with simultaneous AIV and LAD occlusion allows perfusion of oxygenated blood to the distal AIV, which in the setting of an acute myocardial infarction model was capable of rescuing most of the myocardium at risk. PMID- 16345050 TI - Evaluation of the AMPLATZER vascular plug for embolization of peripheral vascular malformations associated with congenital heart disease. AB - OBJECTIVE: The purpose of this study is to evaluate the recently FDA-approved AMPLATZER Vascular Plug in the embolization of vascular lesions associated with congenital heart disease (CHD). BACKGROUND: Fistulas and arteriovenous malformations have been occluded using various devices. The AMPLATZER Vascular Plug is a self-expandable, cylindrical device, used for embolization in the peripheral vasculature. METHODS: A total of 84 vessels in 52 patients with CHD from 11 centers were occluded with 89 AMPLATZER Vascular Plugs, delivered through a coronary guide catheter in various vascular sites, including collaterals, pulmonary arterio-venous and coronary artery fistulas, transhepatic tracts, central shunts, patent ductus arteriosus (PDA), and excluded hepatic vein. Complete vessel occlusion was demonstrated within 10 min in 94% of patients. RESULTS: There was no device embolization, vascular disruption, or procedure related complication. One vascular plug implanted in a large type C PDA required surgical removal followed by PDA ligation, after 5 weeks from successful implant because of significant residual flow through the device. CONCLUSIONS: The AMPLATZER Vascular Plug is an effective transcatheter occlusion device in the embolization of a wide variety of vascular lesions associated with CHD. Based on our early experience, caution should be used when considering the Vascular Plug as a closure device for large PDA. PMID- 16345051 TI - DES: technique still matters. PMID- 16345052 TI - Paclitaxel-eluting stents for the treatment of chronic total coronary occlusions: a strategy of extensive lesion coverage with drug-eluting stents. AB - The recanalization of a chronic total coronary occlusion (CTO) is hampered by a high rate of lesion recurrence. The goal of the present study is to assess the effect of paclitaxel-eluting stents in CTOs in a strategy of extensive stent coverage and the optional use of additional bare metal stents (BMSs). In 82 consecutive patients, a CTO (duration > 2 weeks) was successfully recanalized with implantation of one or more Taxus stents. These patients underwent a repeat angiography after 5.0 +/- 1.5 months and were assessed by quantitative angiography. The patients were compared with 82 clinically and lesion-matched patients from a consecutive series of 148 patients with CTOs treated by BMS in the preceding time period. In 21 of the 82 patients, additional lesions in the target artery not directly related to the original occlusion site were treated with BMSs (hybrid approach). The history of diabetes, extent of coronary artery disease, clinical symptoms, and angiographic features were similar in the Taxus and BMS group. Periprocedural adverse events were 3.3% with Taxus and 3.3% with BMS, but 12 months MACE was significantly lower in the group with exclusive use of Taxus (13.3% vs. 56.7%; P < 0.001), mainly due to a lower target lesion revascularization of 10.0% as compared to 53.4% (P < 0.001). There was only one late reocclusion with Taxus (1.7%) as compared to 21.7% with BMS (P < 0.05). However, in the hybrid group, the MACE rate was considerably higher, with 33.3%. Our data of a 80% reduction of target vessel failure as compared to BMS, with a lower risk of late reocclusions without increased acute adverse events, demonstrate the benefit of paclitaxel-eluting stents in CTOs. However, diffuse atherosclerosis in CTOs should be covered completely by the drug-eluting stents. PMID- 16345053 TI - Intra-atrial tissue plasminogen activator infusion for prosthetic valve thrombosis. AB - Thrombosis is a well-known and life-threatening complication of prosthetic mechanical valves. Therapy typically requires medical thrombolysis or surgical thrombectomy/valve replacement. We report the case of a thrombosed atrioventricular valve in a young boy with complex congenital heart disease, which was successfully treated with direct intra-atrial infusion of recombinant tissue plasminogen activator after failed attempts at systemic therapy. We present this treatment as an alternative to conventional medical therapy. PMID- 16345054 TI - Temporal surveillance using scan statistics. AB - We describe two classes of statistics for testing an arbitrary model of disease incidence over time against an alternative model involving a spike (pulse) superimposed on this background. The statistics are each based on taking the maximum of some function comparing observed and expected numbers of events in a window of width w. One approach applies p-values for scan statistics calculated for a constant background rate to this more general problem. For a fixed window, w, the approach gives a simple formula to determine p-values for retrospective analysis, or to sound an alarm for either continuous or grouped prospective data. The latter application involves a new approximation for the distribution of the maximum number of cases in w consecutive intervals. The second approach based on generalized likelihood ratio tests (GLRTs), sounds an alarm for a higher than anticipated rate of events in a scanning window of fixed length, or for window sizes that lie in a region. GLRTs are constructed for continuous observations, for grouped data, or for a sequence of trials. As for GLRTs used in retrospective evaluations, simulation is required to implement the prospective procedure. For grouped surveillance data, we compare by simulation, operating characteristics of the P-scan with fixed windows (both correctly specified and not), the fixed window GLRT, the variable-window GLRT, and a variant of the CUSUM. The simulations demonstrate a very high correlation between the P-scan and corresponding fixed-window GLRT. PMID- 16345055 TI - Development and psychometric validation of an eating disorder-specific health related quality of life instrument. AB - OBJECTIVE: Health-related quality of life (HRQOL) has been used increasingly as an outcome measure in clinical research. Although the generic quality of life instruments has been used in previous research, disease-specific instruments offer greater sensitivity and responsiveness to change than generic instruments. No such disease-specific instrument is currently available that applies to eating disordered samples. METHOD: The current article reports on the development and validation of the Eating Disorders Quality of Life (EDQOL) instrument, a disease specific HRQOL self-report questionnaire designed for disordered eating patients. RESULTS: The EDQOL demonstrates excellent psychometric properties. CONCLUSION: The application of the EDQOL as an outcome measure in eating disorder research is considered. PMID- 16345056 TI - Estimating measures of diagnostic accuracy when some covariate information is missing. AB - Many biomedical data sets are concerned with relating the result of screening procedure(s) for a clinical event to the occurrence of that event. The effect of risk factors on measures of accuracy such as positive predictive value and negative predictive value is of great interest for clinicians. In this paper we propose a generic approach to estimate these measures of accuracy in the setting where an explanatory model has been fitted to the joint screening and event outcome data but information on one or more risk factors in the model is not available. We refer to these as conditional rates, i.e. rates conditioned on only a subset of risk factors. We argue that, based upon the joint distribution of the event outcome, the screening result and the risk factor occurrence, a formal expression for such a rate can be obtained. This expression is a function of model parameters and thus can be estimated once the model has been fitted. Inference within the Bayesian framework is particularly attractive since simulation based model fitting straightforwardly yields samples from the posterior distribution of any conditional rate of interest. We perform a simulation study to compare these estimated conditional rates with frequently used ad hoc estimates. Differences can be substantial. We also illustrate the proposed methodology to compute conditional positive predictive value for a screening mammography data set. The proposed approach is also applicable when there are multiple diagnostic screening test outcomes. PMID- 16345057 TI - Bayesian sample size calculations in phase II clinical trials using a mixture of informative priors. AB - A number of researchers have discussed phase II clinical trials from a Bayesian perspective. A recent article by Mayo and Gajewski focuses on sample size calculations, which they determine by specifying an informative prior distribution and then calculating a posterior probability that the true response will exceed a prespecified target. In this article, we extend these sample size calculations to include a mixture of informative prior distributions. The mixture comes from several sources of information. For example consider information from two (or more) clinicians. The first clinician is pessimistic about the drug and the second clinician is optimistic. We tabulate the results for sample size design using the fact that the simple mixture of Betas is a conjugate family for the Beta- Binomial model. We discuss the theoretical framework for these types of Bayesian designs and show that the Bayesian designs in this paper approximate this theoretical framework. PMID- 16345058 TI - Confidence intervals for a ratio of binomial proportions based on paired data. AB - Four interval estimation methods for the ratio of marginal binomial proportions are compared in terms of expected interval width and exact coverage probability. Two new methods are proposed that are based on combining two Wilson score intervals. The new methods are easy to compute and perform as well or better than the method recently proposed by Nam and Blackwelder. Two sample size formulas are proposed to approximate the sample size required to achieve an interval estimate with desired confidence level and width. PMID- 16345059 TI - The implications of publication bias for meta-analysis' other parameter. AB - Perhaps the greatest threat to the validity of a meta-analysis is the possibility of publication bias, where studies that are interesting or statistically significant are more likely to be published than those with less encouraging results. In particular, there is the concern that this bias might be 'one-sided', where studies indicating that the treatment is beneficial have a greater probability of publication. The impact that this type of bias has on the estimate of treatment effect has received a great deal of attention but this also has implications for estimates of between-study variance. Using step functions to model the bias it can be demonstrated that it is impossible to make generalizations concerning how we should revise estimates of between-study variance when presented with the possibility of publication bias. To determine this, assumptions must be made concerning the form that the bias takes, which is unknown in practice. PMID- 16345060 TI - Percutaneous closure of an aorta to left atrium fistula with an Amplatzer duct occluder. AB - We describe percutaneous closure of an unusual paravalvular leak, with a communication from the noncoronary cusp of the aortic valve to the left atrium adjacent to a St. Jude mitral valve prosthesis, in the absence of an infection. The patient presented with worsening dyspnea and edema. The anatomic location of the valve annulus adjacent to the noncoronary cusp underlies the etiology of this communication. Outcomes from surgical repair are associated with high mortality. We performed percutaneous closure of the defect, using general anesthesia and transesophageal echo guidance. An Amplatzer duct occluder was placed in the fistula, with immediate and complete closure of the shunt. While no devices specific for closure of paravalvular leaks have been designed, growing experience with a variety of devices and the use of echo guidance have allowed successful therapy in many complex cases. PMID- 16345061 TI - Predicting the lung cancer burden: Accounting for selection of the patients with respect to general population mortality. AB - Incidence, survival, prevalence and mortality are the elements of the cancer burden. The cancer burden is thus contributed by the same individuals from the diagnosis of cancer (incidence) until possible death from cancer (mortality). It would therefore be natural that predictions of future cancer burden, those needed by health administration, for example, could be based on individual data. This paper presents a new model for estimating future cancer burden, a model where individuals are followed from birth to death with or without a diagnosis of cancer. The model can formally be expressed as a summation of log-likelihoods of getting cancer and of surviving the cancer and other causes of death until death or censoring. The new model is illustrated with data of Finnish males with or without a diagnosis of lung cancer in calendar period 1987-1997. Incidence is modelled with a general age-cohort model with a drift and survival from cancer with a parametric mixture model. In a model for survival from other causes of death, selection of patients with respect to general population mortality is accounted for. Future cancer burden is illustrated with short-term predictions of prevalence and mortality. PMID- 16345062 TI - Expression of two temporally distinct microglia-related gene clusters after spinal cord injury. AB - The dual role of microglia in cytotoxicity and neuroprotection is believed to depend on the specific, temporal expression of microglial-related genes. To better clarify this issue, we used high-density oligonucleotide microarrays to examine microglial gene expression after spinal cord injury (SCI) in rats. We compared expression changes at the lesion site, as well as in rostral and caudal regions after mild, moderate, or severe SCI. Using microglial-associated anchor genes, we identified two clusters with different temporal profiles. The first, induced by 4 h postinjury to peak between 4 and 24 h, included interleukin-1beta, interleukin-6, osteopontin, and calgranulin, among others. The second was induced 24 h after SCI, and peaked between 72 h and 7 days; it included C1qB, Galectin-3, and p22(phox). These two clusters showed similar expression profiles regardless of injury severity, albeit with slight decreases in expression in mild or severe injury vs. moderate injury. Expression was also decreased rostral and caudal to the lesion site. We validated the expression of selected cluster members at the mRNA and protein levels. In addition, we demonstrated that stimulation of purified microglia in culture induces expression of C1qB, Galectin-3, and p22(phox). Finally, inhibition of p22(phox) activity within microglial cultures significantly suppressed proliferation in response to stimulation, confirming that this gene is involved in microglial activation. Because microglial-related factors have been implicated both in secondary injury and recovery, identification of temporally distinct clusters of genes related to microglial activation may suggest distinct roles for these groups of factors. PMID- 16345063 TI - Histological identification of syphilis in pre-Columbian England. AB - Microscopic analyses served to complement the macroscopic identification of venereal syphilis in two of four pre-Columbian skeletons from the site Hull Magistrates Court in England. Diagnosis was based on parameters presented by Schultz ([1994] Origin of Syphilis in Europe, Toulon: Centre Archaeologique du Var, p. 63-67; [2001] Yrbk. Phys. Anthropol. 44:106-147; [2003] Identification of Pathological Conditions in Human Remains, New York: Academic Press, p. 73-109), which characterized venereal syphilis at a histological level. Observation of the microarchitecture of these samples allowed a more comprehensive approach to identification of the disease (processes). In most samples, Polsters and Grenzstreifen (or remnants of such structures) could be identified, suggesting the presence of a chronic, inflammatory disease such as venereal syphilis. Sinous lacunae were also observed in all histological samples, pointing to lytic activity (osteitis). The combination of both proliferative and destructive processes is pathognomonic for syphilis, and histological analyses provided a more accurate diagnosis of this infectious disease in these four individuals. As a result, the histological evidence suggests that venereal syphilis was present in England prior to 1492. This secondary form of evidence supports the macroscopic identification of the disease, and shows the power of a multimethodological approach to paleopathological diagnoses. PMID- 16345064 TI - Distribution of postpartum amenorrhea in rural Bangladeshi women. AB - Previous studies of postpartum amenorrhea (PPA) demonstrated distinct subgroups of women with short and long durations of amenorrhea. This phenomenon was attributed to cases where breastfeeding is absent because of pregnancy loss or infant death, or confusion of postpartum bleeding with resumption of menses. We explored these ideas using data from an 11-month prospective study in Bangladesh in which 858 women provided twice-weekly interviews and urine specimens for up to 9 months; 300 women were observed while experiencing PPA. The resulting exact, interval-censored, or right-censored durations were used to estimate parameters of two-component mixture models. A mixture of two Weibull distributions provided the best fit to the observations. The long-duration subgroup made up 84% (+/- 4% SE) of the population, with a mean duration of 457 (+/- 31) days. The short duration subgroup had a mean duration of 94 (+/- 17) days. Three covariates were associated with the duration of PPA: women whose husbands had high-wage employment had a greater probability of falling in the short-duration subgroup; women in the long-duration subgroup whose husbands seasonally migrated had shorter periods of PPA within the subgroup; and mothers in the short-duration subgroup who gave birth during the monsoon season experienced a shortened duration of PPA within the subgroup. We conclude that the bimodal distribution of PPA reflects biological or behavioral heterogeneity rather than shortcomings of data collection. PMID- 16345065 TI - High-resolution X-ray computed tomography scanning of primate copulatory plugs. AB - In this study, high-resolution computed tomography X-ray scanning was used to scan ring-tailed lemur (Lemur catta) copulatory plugs. This method produced accurate measures of plug volume and surface area, but was not useful for visualizing plug internal structure. Copulatory plug size was of interest because it may relate to male fertilization success. Copulatory plugs form from coagulated ejaculate, and are routinely displaced in this species by the penis of a subsequent mate during copulation (Parga [2003] Int. J. Primatol. 24:889-899). Because one potential function of these plugs may be to preclude or delay other males' successful insemination of females, we tested the hypothesis that larger plugs are more difficult for subsequent males to displace. Plugs were collected opportunistically upon displacement during data collection on L. catta mating behavior on St. Catherines Island, Georgia (USA) during two subsequent breeding seasons. Copulatory plugs exhibited a wide range of volumes: 1,758-5,013.6 mm3 (n = 9). Intraindividual differences in plug volume were sometimes greater than interindividual differences. Contrary to predictions, larger plugs were not more time-consuming for males to displace via penile intromission during copulation. Nor were plugs with longer vaginal residence times notably smaller than plugs with shorter residence times, as might be expected if plugs disintegrate while releasing sperm (Asdell [1946] Patterns of Mammalian Reproduction; Ithaca: Comstock). We found a significant inverse correlation between number of copulatory mounts leading to ejaculation and copulatory plug volume. This may indicate that if males are sufficiently sexually aroused to reach ejaculation in fewer mounts, they tend to produce ejaculates of greater volume. PMID- 16345066 TI - Regional, social, and evolutionary perspectives on treponemal infection in the Southeastern United States. AB - The origin and geographic distribution of syphilis, a form of treponemal infection, have long been regarded as among the most important medical riddles of prehistoric and historic disease evolution. In this study, we expand on previous discussions of the origin, evolution, and relationship of treponemal infections as they occur in the prehistoric southeastern United States. Individuals from 25 skeletal series (n = 2,410 individuals) were examined for cranial and dental lesions characteristic of treponemal infection. They lived between the Archaic period (8000-1000 BC) and protohistoric period (AD 1500-1600), and in physiographic zones from the coast to the mountains of Alabama, Florida, North Carolina, and Tennessee. Radial cranial scars were found for 47 individuals, but none of the four cases of dental lesions could be attributed to congenital syphilis. Differences in frequency of cranial lesions by region were minimal, with the least number of cases found for the mountains, but the frequency of positive cases tended to increase through time. It is suggested that increasing population density and changing behaviors, rather than novel strains of the treponemal pathogen, are responsible for the chronological increase in the frequency of positive cases. PMID- 16345067 TI - Life in the slow lane revisited: ontogenetic separation between chimpanzees and humans. AB - This study investigates the evolution of human growth by analyzing differences in body mass growth trajectories among three populations: the Ache of eastern Paraguay, the US (NHANES, 1999-2000), and captive chimpanzees. The relative growth statistic "A" from the mammalian growth law is allowed to vary with age and proves useful for comparing growth across different ages, populations, and species. We demonstrate ontogenetic separation between chimpanzees and humans, and show that interspecific differences are robust to variable environmental conditions. The human pattern of slow growth during the lengthened period from weaning to the beginning of the adolescent growth spurt is found among the Ache (low energy availability and high disease load) and also in the US (high energy availability and low disease load). The human growth pattern contrasts with that of the chimpanzee, where absolute growth rates and relative "A" values are faster and less prolonged. We suggest that selection has acted to decrease human growth rates to allow more time for increased cognitive development with lower body maintenance costs. PMID- 16345068 TI - Masseter electromyography during chewing in ring-tailed lemurs (Lemur catta). AB - We examined masseter recruitment and firing patterns during chewing in four adult ring-tailed lemurs (Lemur catta), using electromyography (EMG). During chewing of tougher foods, the working-side superficial masseter tends to show, on average, 1.7 times more scaled EMG activity than the balancing-side superficial masseter. The working-side deep masseter exhibits, on average, 2.4 times the scaled EMG activity of the balancing-side deep masseter. The relatively larger activity in the working-side muscles suggests that ring-tailed lemurs recruit relatively less force from their balancing-side muscles during chewing. The superficial masseter working-to-balancing-side (W/B) ratio for lemurs overlaps with W/B ratios from anthropoid primates. In contrast, the lemur W/B ratio for the deep masseter is more similar to that of greater galagos, while both are significantly larger than W/B ratios of anthropoids. Because ring-tailed lemurs have unfused and hence presumably weaker symphyses, these data are consistent with the symphyseal fusion muscle recruitment hypothesis stating that symphyseal fusion in anthropoids provides increased strength for resisting forces created by the balancing-side jaw muscles during chewing. Among the masseter muscles of ring-tailed lemurs, the working-side deep masseter peaks first on average, followed in succession by the balancing-side deep masseter, balancing-side superficial masseter, and finally the working-side superficial masseter. Ring-tailed lemurs are similar to greater galagos in that their balancing-side deep masseter peaks well before their working-side superficial masseter. We see the opposite pattern in anthropoids, where the balancing-side deep masseter peaks, on average, after the working-side superficial masseter. This late activity of the balancing-side deep masseter in anthropoids is linked to lateral-transverse bending, or wishboning, of their mandibular symphyses. Subsequently, the stresses incurred during wishboning are hypothesized to be a proximate reason for strengthening, and hence fusion, of the anthropoid symphysis. Thus, the absence of this muscle-firing pattern in ring tailed lemurs with their weaker, unfused symphyses provides further correlational support for the symphyseal fusion late-acting balancing-side deep masseter hypothesis linking wishboning and symphyseal strengthening in anthropoids. The early peak activity of the working-side deep masseter in ring-tailed lemurs is unlike galagos and most similar to the pattern seen in macaques and baboons. We hypothesize that this early activity of the working-side deep masseter moves the lower jaw both laterally toward the working side and vertically upward, to position it for the upcoming power stroke. From an evolutionary perspective, the differences in peak firing times for the working-side deep masseter between ring tailed lemurs and greater galagos indicate that deep masseter firing patterns are not conserved among strepsirrhines. PMID- 16345069 TI - Morphometric cranial identity of prehistoric Malawians in the light of sub Saharan African diversity. AB - Little has been described of the Holocene populations of South-Central Africa, despite the region demonstrating major subsistence shifts relating to dispersals of agriculturalists at least 2,000 years ago. Seven sites with associated human skeletal remains were selected. Hora, Chencherere, Fingura, and Mtuzi represent the Middle Holocene (2,000-5,000 years ago), and Phwadze, Mtemankhokwe, and Nkudzi Bay represent the Late Holocene and the arrival of agriculturalists between 500-2,000 years ago. Focusing on the identity of Hora and Chencherere specimens, two questions were addressed: are the various Holocene Malawians similar to each other, or do they suggest morphological change over time? What modern populations are closest to the prehistoric specimens? The archaeological sample was compared to modern sub-Saharan Africans from four regions, plus a historic Khoi-San foraging group. Factor analyses were performed in order to identify complex patterns of variation in metric traits of the skull. According to the results, prehistoric Malawians showed only slight differences between the Late and Middle Holocene, suggesting a population change without any major discontinuity. Later Stone Age skulls did not exclusively show similarities with the Khoi-San, as they frequently fit well within the variation of modern Bantu speaking groups, especially West-Central Africa. Therefore, we reject the hypothesis that Middle Holocene South-Central Africans have an exclusively Khoi San ancestry, and support an alternative hypothesis that both Middle and Late Holocene groups share a common biological heritage originating in West-Central Africa in earlier times. PMID- 16345070 TI - Effect of ethnicity and sex on the growth of the axial and appendicular skeleton of children living in a developing country. AB - Bones in the axial and appendicular skeletons exhibit heterogeneous growth patterns between different ethnic and sex groups. However, the influence of this differential growth on the expression of bone mineral content is not yet established. The aims of the present study were to investigate: 1) whether there are ethnic and sex differences in axial and appendicular dimensions of South African children; and 2) whether regional segment length is a better predictor of bone mass than stature. Anthropometric measurements of stature, weight, sitting height, and limb lengths were taken on 368 black and white, male and female 9 year-old children. DXA (dual-energy x-ray absorptiometry) scans of the distal ulna, distal radius, and hip and lumbar spine were also obtained. Analyses of covariance were performed to assess differences in limb lengths, adjusted for differences in stature. Multiple regression analyses were used to assess significant predictors of site-specific bone mass. Stature-adjusted means of limb lengths show that black boys have longer legs and humeri but shorter trunks than white boys. In addition, black children have longer forearms than white children, and girls have longer thighs than boys. The regression analysis demonstrated that site-specific bone mass was more strongly associated with regional segment length than stature, but this had little effect on the overall pattern of ethnic and sex differences. In conclusion, there is a differential effect of ethnicity and sex on the growth of the axial and appendicular skeletons, and regional segment length is a better predictor of site-specific bone mass than stature. PMID- 16345071 TI - Cranial ontogeny of Papio baboons (Papio hamadryas). AB - Cranial form in subspecies of Papio baboons (Papio hamadryas) varies in relation to size, geography, and sex. However, knowledge about this variation is based mainly on adults, precluding direct assessments of the evolutionary factors that are ultimately responsible for adult shape variation. Consequently, this study tests hypotheses about the development of size and shape differences among subspecies of Papio baboons, anticipating limited evolutionary divergences in the ontogenetic pathways leading to adult endpoints. Geometric morphometric and bivariate allometric analyses are used to explore developmental size and shape variation. Allometric scaling in adult Papio baboons occurs because both sexes and all subspecies follow similar developmental pathways to a variety of adult forms. However, complex allometry contributes to form differences, producing potentially important shape differences that emerge during development. Modest shape differences that are statistically independent of size distinguish chacma baboons (P. h. ursinus) from other forms. A small-headed subspecies, the Kinda baboon (P. h. kindae), also presents a distinctive ontogeny, and may provide insights into the evolution of size change in this species. Variation among subspecies that is statistically independent of size involves the rostrum, zygomatic breadths, and cranial flexion. These features may be related to diet, but the precise biomechanical correlates of baboon form variation remain unclear. PMID- 16345072 TI - Phylogenetic analyses of dimorphism in primates: evidence for stronger selection on canine size than on body size. AB - Phylogenetic comparative methods were used to analyze the consequences of sexual selection on canine size and canine size dimorphism in primates. Our analyses of previously published body mass and canine size data revealed that the degree of sexual selection is correlated with canine size dimorphism, as well as with canine size in both sexes, in haplorhine but not in strepsirrhine primates. Consistent with these results, male and female canine size was found to be highly correlated in all primates. Since canine dimorphism and canine size in both sexes in haplorhines were found to be not only related to mating system but also to body size and body size dimorphism (characters which are also subject to or the result of sexual selection), it was not apparent whether the degree of canine dimorphism is the result of sexual selection on canine size itself, or whether canine dimorphism is instead a consequence of selection on body size, or vice versa. To distinguish among these possibilities, we conducted matched-pairs analyses on canine size after correcting for the effects of body size. These tests revealed significant effects of sexual selection on relative canine size, indicating that canine size is more important in haplorhine male-male competition than body size. Further analyses showed, however, that it was not possible to detect any evolutionary lag between canine size and body size, or between canine size dimorphism and body size dimorphism. Additional support for the notion of special selection on canine size consisted of allometric relationships in haplorhines between canine size and canine size dimorphism in males, as well as between canine size dimorphism and body size dimorphism. In conclusion, these analyses revealed that the effects of sexual selection on canine size are stronger than those on body size, perhaps indicating that canines are more important than body size in haplorhine male-male competition. PMID- 16345073 TI - Crystal structure of glyceraldehyde-3-phosphate dehydrogenase from Plasmodium falciparum at 2.25 A resolution reveals intriguing extra electron density in the active site. AB - The crystal structure of D-glyceraldehyde-3-phosphate dehydrogenase (PfGAPDH) from the major malaria parasite Plasmodium falciparum is solved at 2.25 A resolution. The structure of PfGAPDH is of interest due to the dependence of the malaria parasite in infected human erythrocytes on the glycolytic pathway for its energy generation. Recent evidence suggests that PfGAPDH may also be required for other critical activities such as apical complex formation. The cofactor NAD(+) is bound to all four subunits of the tetrameric enzyme displaying excellent electron densities. In addition, in all four subunits a completely unexpected large island of extra electron density in the active site is observed, approaching closely the nicotinamide ribose of the NAD(+). This density is most likely the protease inhibitor AEBSF, found in maps from two different crystals. This putative AEBSF molecule is positioned in a crucial location and hence our structure, with expected and unexpected ligands bound, can be of assistance in lead development and design of novel antimalarials. PMID- 16345074 TI - Prediction of protein secondary structure content using amino acid composition and evolutionary information. AB - Knowing protein structure and inferring its function from the structure are one of the main issues of computational structural biology, and often the first step is studying protein secondary structure. There have been many attempts to predict protein secondary structure contents. Previous attempts assumed that the content of protein secondary structure can be predicted successfully using the information on the amino acid composition of a protein. Recent methods achieved remarkable prediction accuracy by using the expanded composition information. The overall average error of the most successful method is 3.4%. Here, we demonstrate that even if we only use the simple amino acid composition information alone, it is possible to improve the prediction accuracy significantly if the evolutionary information is included. The idea is motivated by the observation that evolutionarily related proteins share the similar structure. After calculating the homolog-averaged amino acid composition of a protein, which can be easily obtained from the multiple sequence alignment by running PSI-BLAST, those 20 numbers are learned by a multiple linear regression, an artificial neural network and a support vector regression. The overall average error of method by a support vector regression is 3.3%. It is remarkable that we obtain the comparable accuracy without utilizing the expanded composition information such as pair coupled amino acid composition. This work again demonstrates that the amino acid composition is a fundamental characteristic of a protein. It is anticipated that our novel idea can be applied to many areas of protein bioinformatics where the amino acid composition information is utilized, such as subcellular localization prediction, enzyme subclass prediction, domain boundary prediction, signal sequence prediction, and prediction of unfolded segment in a protein sequence, to name a few. PMID- 16345075 TI - NTP-CERHR Expert Panel Report on the reproductive and developmental toxicity of styrene. PMID- 16345076 TI - The Histone Database: a comprehensive resource for histones and histone fold containing proteins. AB - The Histone Database is a curated and searchable collection of full-length sequences and structures of histones and nonhistone proteins containing histone like folds, compiled from major public databases. Several new histone fold containing proteins have been identified, including the huntingtin-interacting protein HYPM. Additionally, based on the recent crystal structure of the Son of Sevenless protein, an interpretation of the sequence analysis of the histone fold domain is presented. The database contains an updated collection of multiple sequence alignments for the four core histones (H2A, H2B, H3, and H4) and the linker histones (H1/H5) from a total of 975 organisms. The database also contains information on the human histone gene complement and provides links to three dimensional structures of histone and histone fold-containing proteins. The Histone Database is a comprehensive bioinformatics resource for the study of structure and function of histones and histone fold-containing proteins. The database is available at http://research.nhgri.nih.gov/histones/. PMID- 16345077 TI - Morphology and DNA degeneration during autoschizic cell death in bladder carcinoma T24 cells induced by ascorbate and menadione treatment. AB - Feulgen and actin-phalloidin staining as well as gel electrophoresis have been employed in conjunction with cell ultrastructure to describe the effects of 1-, 2 , and 4-hr ascorbate (VC), menadione (VK(3)), and ascorbate:menadione (VC:VK(3)) treatments on the T24 human bladder carcinoma cell line. T24 cells exposed to VC alone display blebs and other superficial membrane defects related to membrane alterations and to superficial cytoskeleton changes. VK(3) treatment damages the cell nucleus and organelles, leads to the redistribution of the organelles in the perikaryon as a consequence of cytoskeletal damage, and results in cytoplasmic self-excisions. After VC:VK(3) treatment, the cells show exaggerated alterations characteristic of each vitamin treatment alone, including damaged mitochondria, self-excision of organelle-free pieces of cytoplasm, and extrusion of the perikaryon containing a nucleus surrounded by the damaged organelles. The nuclear envelope appears intact and contains chromatin that decondenses and dissipates. During the cellular demise that concludes with apparent karyolysis, the cells significantly decrease their size and alter their shape. However, the cisterns of rough endoplasmic reticulum are undamaged, but may become dilated. Since the cellular phenomena leading to cell death differ morphologically from apoptosis and necrosis, but entail self-cutting without nuclear bodies, this new form of cell death was called autoschizis. In addition, gel electrophoresis and Feulgen staining demonstrate that autoschizis is accompanied by random DNA degeneration. PMID- 16345078 TI - Three-dimensional fiber architecture of the nonpregnant human uterus determined ex vivo using magnetic resonance diffusion tensor imaging. AB - The global muscle and collagen fiber orientation in the human uterus has been analyzed hitherto by various standard microscopic techniques. However, no widely accepted model of the fiber architecture of the myometrium could be acquired. The purpose of the present study was to investigate the uterus by magnetic resonance (MR) diffusion tensor imaging (DTI) in a 3D macroscopic approach. Ex vivo MR DTI measurements were performed on five uteri from nonpregnant patients. The main diffusion directions reflecting the orientation of directional structures in the examined tissues were determined from diffusion-weighted spin-echo measurements. A fiber tracking algorithm was used to extrapolate the fiber architecture. The method was validated against histological slides and indirectly through the analysis of leiomyomas, which exhibit less anisotropy than normal myometrium. Significant anisotropy was found in most regions of all examined nonpregnant human uteri. But only two systems of fibers were found running circularly along the intramural part of the uterine tubes. They merged caudally and built a close fitting envelope of circular layers around the uterine cavity. On the cervix, circular fibers were observed in the outer part as well as mostly longitudinal fibers in the inner part. These results confirm the existence of directional structures in the complex fiber architecture of the human uterus. They also indicate that MR DTI is a beneficial and complementary tool to standard microscopic techniques to determine the intrinsic fiber architecture in human organs. PMID- 16345079 TI - Crystal structure of antigen TpF1 from Treponema pallidum. PMID- 16345080 TI - Asymmetrically functional surface properties on biocompatible phospholipid polymer membrane for bioartificial kidney. AB - To obtain a bioartificial kidney composed of a porous polymer membrane and renal cells, a polysulfone (PSf) membrane (PSM) blended with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer was prepared. The PSM flat membrane with a porous structure could be prepared from the polymer blend containing 1 wt % of the MPC polymer in PSf by the phase inversion technique in a dry-wet process. Asymmetrical surface properties were observed on both sides of the membrane surfaces. That is, the sponge layer formed at the substrate-contacting surface of the membrane had 10-20 microm pores, but the pores in the micrometer range could not be observed for a skin layer formed at the air-contacting surface of the membrane. At the sponge layer surface, the MPC unit composition was 7 times larger than that at the skin layer surface. The amount of proteins adsorbed on the surface corresponded to the MPC unit composition. On the skin layer, a small amount of adsorbed proteins and platelet adhesion could be suppressed compared with those on the sponge layer. However, the skin layer had a moderate protein adsorption, so it showed a sufficient cytocompatibility to enable renal tubule epithelial cells to adhere and proliferate in the membrane. Thus, it functioned well as a renal tubule. Therefore, because of both its hemocompatibility and cytocompatibility, we could conclude that the PSM membrane is useful for as a renal tubule device for a bioartificial kidney. PMID- 16345081 TI - Alginate-PLL microencapsulation: effect on the differentiation of embryonic stem cells into hepatocytes. AB - The emergence of hepatocyte based clinical and pharmaceutical technologies, has been limited by the absence of a stable hepatocyte cell source. Embryonic stem cells may represent a potential solution to this cell source limitation problem since they are highly proliferative, renewable, and pluripotent. Although many investigators have described techniques to effectively differentiate stem cells into a variety of mature cell lineages, their practicality is limited by: (1) low yields of fully differentiated cells, (2) absence of large scale processing considerations, and (3) ineffective downstream enrichment protocols. Thus, a differentiation platform that may be modified to induce and sustain differentiated cell function and scaled to increase differentiated cell yield would improve current stem cell differentiation strategies. Microencapsulation provides a vehicle for the discrete control of key cell culture parameters such as the diffusion of growth factors, metabolites, and wastes. In addition, both cell seeding density and bead composition may be manipulated. In order to assess the feasibility of directing stem cell differentiation via microenvironment regulation, we have developed a murine embryonic stem cell (ES) alginate poly-l lysine microencapsulation hepatocyte differentiation system. Our results indicate that the alginate microenvironment maintains cell viability, is conducive to ES cell differentiation, and maintains differentiated cellular function. This system may ultimately assist in developing scalable stem cell differentiation strategies. PMID- 16345082 TI - Responses of MC3T3-E1 cells to three dental resin-based restorative materials. AB - This study aimed to examine the influences of three dental resin-based restorative materials on cells associated with hard tissue regeneration using osteoblastic MC3T3-E1 cells. A Bis-GMA-based resin composite [Clearfil AP-X (APX)], an MMA-based resin cement [Superbond C&B (SB)], and a resin-modified glass-ionomer [Fuji Ionomer Type II LC (LC)] were tested. A zinc oxide eugenol cement [Super EBA (EBA)] was included in the study for comparison. MC3T3-E1 cells were cultured on set materials for 3, 7, 14, or 21 days. Cell attachment and proliferation were observed by scanning electron microscopy, and mitochondrial dehydrogenase and alkaline phosphatase (ALP) activities of the cells were evaluated. Cell cultures on polystyrene tissue culture dishes served as controls. On APX and SB, cells demonstrated attachment, spreading, and proliferation similar to the controls. In contract, cells adhered and proliferated poorly on LC and EBA. The mitochondrial function and ALP activity of the cells were significantly suppressed (p < 0.05, Scheffe's F test) throughout the experimental period when cultured on LC or EBA, although APX and SB exhibited less inhibition. The results indicate that APX and SB are less toxic to proliferation and differentiation of MC3T3-E1, suggesting that a smaller influence on cementogenesis on these materials can be expected. PMID- 16345083 TI - Transparent micro- and nanopatterned poly(lactic acid) for biomedical applications. AB - The formation of structures in poly(lactic acid) has been investigated with respect to producing areas of regular, superficial features with dimensions comparable to those of cells or biological macromolecules. Nanoembossing, a novel method of pattern replication in polymers, has been used for the production of features ranging from tens of micrometers, covering areas up to 1 cm(2), down to hundreds of nanometers. Both micro- and nanostructures are faithfully replicated. Contact-angle measurements suggest that positive microstructuring of the polymer (where features protrude from the polymer surface) produces a more hydrophilic surface than negative microstructuring. The ability to structure the surface of the poly(lactic acid), allied to the polymer's postprocessing transparency and proven biocompatibility, means that thin films produced in this way will be useful for bioengineers studying the interaction of micro- and nanodimensioned features with biological specimen, with regard to tissue engineering, for example. PMID- 16345084 TI - Single dose vaccine based on biodegradable polyanhydride microspheres can modulate immune response mechanism. AB - This study focuses on the development of single dose vaccines based on biodegradable polyanhydride microspheres that have the unique capability to modulate the immune response mechanism. The polymer system employed consists of copolymers of 1,6-bis(p-carboxyphenoxy)hexane and sebacic acid. Two copolymer formulations that have been shown to provide extended release kinetics and protein stability were investigated. Using tetanus toxoid (TT) as a model antigen, in vivo studies in C3H/HeOuJ mice demonstrated that the encapsulation procedure preserves the immunogenicity of the TT. The polymer itself exhibited an adjuvant effect, enhancing the immune response to a small dose of TT. The microspheres provided a prolonged exposure to TT sufficient to induce both a primary and a secondary immune response (i.e., high antibody titers) with high avidity antibody production, without requiring an additional administration. Antigen-specific proliferation 28 weeks after a single immunization indicated that immunization with the polyanhydride microspheres generated long-lived memory cells and plasma cells (antibody-secreting B cells) that generally do not occur without maturation signals from T helper cells. Furthermore, by altering the vaccine formulation, the overall strength of the T helper type 2 immune response was selectively diminished, resulting in a balanced immune response, without reducing the overall titer. This result is striking, considering free TT induces a T helper type 2 immune response, and has important implications for developing vaccines to intracellular pathogens. The ability to selectively tune the immune response without the administration of additional cytokines or noxious adjuvants is a unique feature of this delivery vehicle that may make it an excellent candidate for vaccine development. PMID- 16345085 TI - Effect of wall growth on the kinetic modeling of nitrite oxidation in a CSTR. AB - A simple kinetic model was developed for describing nitrite oxidation by autotrophic aerobic nitrifiers in a continuous stirred tank reactor (CSTR), in which mixed (suspended and attached) growth conditions prevail. The CSTR system was operated under conditions of constant nitrite feed concentration and varying volumetric flow rates. Experimental data from steady-state conditions in the CSTR system and from batch experiments were used for the determination of the model's kinetic parameters. Model predictions were verified against experimental data obtained under transient operating conditions, when volumetric flow rate and nitrite feed concentration disturbances were imposed on the CSTR. The presented kinetic modeling procedure is quite simple and general and therefore can also be applied to other mixed growth biological systems. PMID- 16345086 TI - Alkaline-treated poly(epsilon-caprolactone) films: degradation in the presence or absence of fibroblasts. AB - In the first stage, we observed the study of the degradation behavior of alkaline treated poly(epsilon-caprolactone) (PCL) in two biologically-related media: phosphate buffered saline (PBS) and Dulbecco's modified Eagle's medium (DMEM) for 18 months, finding a much accelerated degradation in the last one. As expected, the degradation in the presence of cells is much pronounced even considering that the study is limited to 6 months. The characterization of the degraded substrates by chemiluminescence (CL) allows to explain the modifications of the substrate and their relations with transitory oxidative stress phenomena described in the fibroblasts seeded onto the PCL membranes. PMID- 16345087 TI - Human neutrophils reaction to the biodegraded nano-hydroxyapatite/collagen and nano-hydroxyapatite/collagen/poly(L-lactic acid) composites. AB - The impact of biodegraded nano-hydroxyapatite/collagen (nHAC) composite and nano hydroxyapatite/collagen/poly(L-lactic acid) (nHAC/PLA) scaffold composite on neutrophils reaction was evaluated in vitro. Neutrophils were separated from human peripheral blood of healthy subjects. The nHAC and nHAC/PLA materials were immersed in the D-Hanks' Balanced Salt Solution (D-HBSS) for 1 day, 7 days and 2, 4, 8 weeks (37 degrees C) as testing solution, which mixed with the neutrophils for 1 h. Both of the nHAC and nHAC/PLA materials were shown the same cell survival rate as blank control, but the lactate dehydrogenase (LDH) and tumor necrosis factor alpha (TNF-alpha) released from the neutrophils were increased significantly after the 2 weeks in nHAC sample. The possible reason relied on the high concentration of calcium due to the quick biodegradation of the nHAC material. Before 2 weeks, the LDH value of nHAC/PLA is higher than that of nHAC sample that corresponded to the initial PLA degradation in vitro. This study provided the biocompatibility test of neutrophils other than common methods, such as osteoblastic cells for biomimetic materials. Moreover, it demonstrated the calcium concentration stimulating effect for cytokine release from neutrophils. PMID- 16345088 TI - A rapid microassay to evaluate enzymatic hydrolysis of lignocellulosic substrates. AB - Current attempts to produce ethanol from lignocellulosic biomass are focused on the optimization of pretreatment to reduce substrate recalcitrance and the improvement of enzymes for hydrolysis of the cellulose and hemicellulose components to produce fermentable sugars. Research aimed at optimizing both aspects of the bioconversion process involves assessment of the effects of multiple variables on enzyme efficiency, resulting in large factorial experiments with intensive assay requirements. A rapid assay for lignocellulose hydrolysis has been developed to address this need. Pretreated lignocellulose is formed into handsheets, which are then used to prepare small disks that are easily dispensed into microtiter plates. The hydrolysis of cellulose to glucose is estimated using an enzyme-coupled spectrophotometric assay. Using disks prepared from ethanol organosolv pretreated yellow poplar, it is shown that the assay generates data comparable with those produced by hydrolysis of pretreated yellow poplar pulp in Erlenmeyer flasks, followed by HPLC analysis of glucose. The assay shows considerable time and cost benefits over the standard assay protocol and is applicable to a broad range of lignocellulosic substrates. PMID- 16345089 TI - Three-dimensional nanofibrillar surfaces covalently modified with tenascin-C derived peptides enhance neuronal growth in vitro. AB - Current methods to promote growth of cultured neurons use two-dimensional (2D) glass or polystyrene surfaces coated with a charged molecule (e.g. poly-L-lysine (PLL)) or an isolated extracellular matrix (ECM) protein (e.g. laminin-1). However, these 2D surfaces represent a poor topological approximation of the three-dimensional (3D) architecture of the assembled ECM that regulates neuronal growth in vivo. Here we report on the development of a new 3D synthetic nanofibrillar surface for the culture of neurons. This nanofibrillar surface is composed of polyamide nanofibers whose organization mimics the porosity and geometry of the ECM. Neuronal adhesion and neurite outgrowth from cerebellar granule, cerebral cortical, hippocampal, motor, and dorsal root ganglion neurons were similar on nanofibers and PLL-coated glass coverslips; however, neurite generation was increased. Moreover, covalent modification of the nanofibers with neuroactive peptides derived from human tenascin-C significantly enhanced the ability of the nanofibers to facilitate neuronal attachment, neurite generation, and neurite extension in vitro. Hence the 3D nanofibrillar surface provides a physically and chemically stabile cell culture surface for neurons and, potentially, an exciting new opportunity for the development of peptide-modified matrices for use in strategies designed to encourage axonal regrowth following central nervous system injury. PMID- 16345090 TI - Organic overlayer model of a dental composite analyzed by laser desorption postionization mass spectrometry and photoemission. AB - Some dental composites consist of a polymerizable resin matrix bound to glass filler particles by silane coupling agents. The resin in these composites includes bisphenol A diglycidyl methacrylate (Bis-GMA) as well as other organic components. Silane coupling agents such as 3-(trimethoxysilyl) propyl methacrylate (MPS) have been used to improve the mechanical properties of the dental composites by forming a covalent bond between the glass filler particles and the resin. These resin-glass composites undergo material property changes during exposure to the oral environment, but degradation studies of the commercial composites are severely limited by their chemical complexity. A simplified model of the dental composite has been developed, which captures the essential chemical characteristics of the filler particle-silane-resin interface. This model system consists of the resin matrix compound Bis-GMA covalently bound via a methacryloyl overlayer to amorphous silicon oxide (SiO2) surface via a siloxane bond. Scanning electron microscopy shows the porous characteristic and elemental composition of the SiO2 film, which approximately mimics that of the glass filler particles used in dental composites. LDPI MS and XPS verify the chemistry and morphology of the Bis-GMA-methacryloyl overlayer. Preliminary results demonstrate that LDPI MS will be able to follow the chemical processes resulting from aging Bis-GMA-methacryloyl overlayers aged in water, artificial saliva, or other aging solutions. PMID- 16345092 TI - Comparison of endothelial cells grown on different stent materials. AB - We compared the behavior of endothelial cells grown on stent materials. Human umbilical vein endothelial cells (HUVEC) were seeded (200 or 800 cells/mm(2)) onto different metallic sheets, including 316 stainless steel (low carbon; 316LSS), nitinol, and 316LSS coated with TiN or TiO(2). Cells seeded onto tissue culture-treated polystyrene dish coated with gelatin were used as controls. Forty eight hours later, the cells were examined by Western blotting, immunofluorescence microscopy, and scanning electron microscopy (SEM). The results showed that for either seeding values, the levels of cellularity on TiN and TiO(2) are comparable or higher, and those on 316LSS and nitinol are lower compared to the controls (p < 0.05). SEM demonstrated that cells are well attached on the metallic surface with various amount of cellular processes. In metals seeded with 800 cells/mm(2), Western blotting showed that the overlying cells expressed less amounts of endothelial nitric oxide synthase (eNOS), Von Willebrand factor (VWF), and connexin43 protein compared to the controls (p < 0.05). Immunofluorescence microscopy confirmed the results of immunoblotting. In conclusion, stent materials affect HUVEC's growth and protein expression profile. Down-regulation of eNOS, VWF, and connexin43 gap junctions is a common phenomenon in the cells growing on the examined metallic materials, suggesting the existence of endothelial dysfunction in the arterial segments containing the stents made of such materials. PMID- 16345091 TI - Manufacture of multimicrotubule chitosan nerve conduits with novel molds and characterization in vitro. AB - Multimicrotubule chitosan conduits (M-conduits) were fabricated using novel molds and a thermal-induced phase-separation technique. Hollow chitosan conduits (H conduits) with an inner diameter of 1-5 mm and a wall thickness of 0.2-1.0 mm were made, and then a novel mold composed of a styrofoam insulating pedestal with several holes and a stainless steel cover plate was used to make M-conduits. In brief, corresponding H-conduits were inserted upright into the holes of the styrofoam pedestal, and filled with chitosan solution, then rapidly covered with the precooled stainless steel cover plate, and then placed in a freezer. The styrofoam insulating pedestal enclosing the conduits could reduce the heat transfer through the side wall of the conduits. Gradual phase separation then occurred uniaxially in the presence of a unidirectional temperature gradient from the top end to the bottom end of the chitosan conduits. The phase-separated polymer/solvent systems were then dried in a freeze-dryer. The microtubule diameters were controlled by adjusting the polymer concentration and cooling temperature. In vitro characterization demonstrated that the mold-based multimicrotubule chitosan conduits possessed suitable mechanical strength, microtubule diameter distribution, porosity, swelling, biodegradability, and nerve cell affinity, and so they showed potential for application as nerve tissue engineering scaffolds. PMID- 16345093 TI - Role of phase diagram of membrane formation system in controlling the crystallinity and degradation rate of PLLA membranes. AB - In this work, the theoretical phase diagram of membrane formation system of ethanol, methylene chloride, and poly-L-lactide (PLLA) was studied. On the basis of the phase diagram, particulate and porous membranes, dominated by crystallization and liquid-liquid demixing, respectively, were prepared. Furthermore, degradation of PLLA membranes with particulate, porous, and dense morphologies was performed in phosphate buffered solution (PBS) at 37 degrees C for 168 days and was investigated by mass loss, scanning electron microscopy (SEM), gel permeation chromatography (GPC), and differential scanning calorimetry (DSC). Besides the membrane morphology, a close relationship between the phase behavior of the membrane formation system and the membrane crystallinity was found, which in turn influenced the degradation rate of these membranes significantly. In the case of dense membranes, it showed the lowest initial crystallinity and the greatest rate of mass loss and molecular weight decrease compared with particulate and porous membranes. In contrast, the particulate membranes had the highest crystallinity and the slowest degradation rate in this study. Therefore, the phase diagram of membrane formation system could not only anticipate membrane morphology, but could also control the membrane crystallinity and degradation rate simultaneously. PMID- 16345094 TI - In vitro antibacterial and cytotoxicity assay of multilayered polyelectrolyte functionalized stainless steel. AB - Infection of implanted materials by bacteria constitutes one of the most serious complications following prosthetic and implant surgery. In the present study, a new strategy for confering stainless steel with antibacterial property via the alternate deposition of quaternized polyethylenimine (PEI) or quaternized polyethylenimine-silver complex and poly(acrylic acid) (PAA) was investigated. The success of the deposition of the polyelectrolyte multilayers (PEM) and its chemical nature was investigated by static water contact angle and X-ray photoelectron spectroscopy (XPS), respectively. The antibacterial activity was assessed using Escherichia coli (E. coli, a gram-negative bacterium) and Staphylococcus aureus (S. aureus, a gram-positive bacterium). The inhibition of E. coli and S aureus growth on the surface of functionalized films was clearly shown using the LIVE/DEAD Baclight bacterial viability kits and fluorescence microscopy. The cytotoxicity of the PEM to mammalian cells, evaluated by the MTT assay, was shown to be minimal and long-term antibacterial efficacy can be maintained. These results indicate new possibilities for the use of such easily built and functionalized architectures for the functionalization of surfaces of implanted medical devices. PMID- 16345095 TI - Thermally responsive polymeric micelles self-assembled by amphiphilic polyphosphazene with poly(N-isopropylacrylamide) and ethyl glycinate as side groups: polymer synthesis, characterization, and in vitro drug release study. AB - Thermally responsive amphiphilic poly(N-isopropylacrylamide) (PNIPAm)-grafted polyphosphazene (PNIPAm-g-PPP) was synthesized by stepwise cosubstitution of chlorine atoms on polymer backbones with amino-terminated NIPAm oligomers and ethyl glycinate (GlyEt). Polymer structure was confirmed by FT-IR, (1)H NMR, elemental analysis, and GPC. The thermosensitivity of PNIPAm-g-PPP aqueous solution was investigated by turbidity method. The lower critical solution temperature (LCST) of PNIPAm-g-PPP was observed to be approximately 30 degrees C in water, while it was 24 degrees C in 0.1M PBS (pH 7.4). Micellization behavior of PNIPAm-g-PPP in aqueous solution was characterized by fluorescence probe technique, TEM, and DLS. The critical micelle concentration (CMC), thus, determined was 0.0187 g/L. Both TEM and DLS measurement suggested that the diameter of micelles was approximately 190 nm at 20 degrees C. Diflunisal (DIF) loaded micelles were prepared by dialysis method. In vitro release test at various temperatures was also performed to study the effect of temperature on the drug release profiles. It was demonstrated that DIF release from PNIPAm-g-PPP micelles was slower at the temperature of 37 degrees C than that at 4 degrees C. PMID- 16345096 TI - Effects of the length of crosslink chain on poly(2-hydroxyethyl methacrylate) (pHEMA) swelling and biomechanical properties. AB - Polymers are widely used in medicine for vascular prostheses, bone substitutes, and devices for controlled release. Among these polymers, poly(2-hydroxyethyl methacrylate) (pHEMA) is the most employed. To confer particular properties, pHEMA can be copolymerized with other monomers or in the presence of plasticizers or crosslinking agents. The influence of the length of crosslink chains on swelling, surface rugosity, hardness, and stiffness of crosslinked pHEMA were studied by several techniques, including fractal analysis and AFM. Four crosslinking agents (divinyl benzene, DVB; ethylene glycol dimethacrylate, EGDMA; tetraethylene glycol diacrylate, TEGDA; and polyethylene glycol diacrylate, PEGDA) were added to the bulk polymerization mixture. Only linear and PEGDA-pHEMA presented a significant decrease in surface roughness confirmed by fractal analysis. Differences in hardness and biomechanical properties were evidenced on dried polymers but the highest differences were exhibited for hydrated pHEMA. Correlations between the length of the crosslink chain and hardness or stiffness of hydrated crosslinked pHEMA were evidenced. TEGDA and PEGDA appeared to be the two most suitable crosslinking agents for controlled release of bioactive molecules in bone. PMID- 16345097 TI - Silicon inhibition effects on the polymerase chain reaction: a real-time detection approach. AB - In the miniaturization of biochemical analysis systems, biocompatibility of the microfabricated material is a key feature to be considered. A clear insight into interactions between biological reagents and microchip materials will help to build more robust functional bio-microelectromechanical systems (BioMEMS). In the present work, a real-time polymerase chain reaction (PCR) assay was used to study the inhibition effects of silicon and native silicon oxide particles on Hepatitis B Virus (HBV) DNA PCR amplification. Silicon nanoparticles with different surface oxides were added into the PCR mixture to activate possible interactions between the silicon-related materials and the PCR reagents. Ratios of silicon nanoparticle surface area to PCR mixture volume (surface to volume ratio) varied from 4.7 to 235.5 mm2/microL. Using high speed centrifugation, the nanoparticles were pelleted to tube inner surfaces. Supernatant extracts were then used in subsequent PCR experiments. To test whether silicon materials participated in amplifications directly, in some cases, entire PCR mixture containing silicon nanoparticles were used in amplification. Fluorescence histories of PCR amplifications indicated that with the increase in surface to volume ratio, amplification efficiency decreased considerably, and within the studied ranges, the higher the particle surface oxidation, the stronger the silicon inhibition effects on PCR. Adsorption of Taq polymerase (not nucleic acid) on the silicon related material surface was the primary cause of the inhibition phenomena and silicon did not participate in the amplification process directly. PMID- 16345098 TI - Effects of 58S sol-gel glasses on the temporal expression of bone markers during mouse osteoblastic differentiation. AB - Previous studies have shown that bioactive glasses can support osteoblastic growth and differentiation in vitro as well as in vivo. More recently, a new category of sol-gel glasses has been developed with enhanced bioactivity and open pores enclosed in a mesoporous matrix. In our study, we investigated the effect of 58S sol-gel glasses on the growth and differentiation of mouse calvaria osteoblasts. Two types of granules were used: 58S sol-gel granules and 60S inert glasses used as control. Phase contrast microscopy showed that cells proliferated and formed mineralized bone nodules in both cultures. However, this phenomenon occurred earlier and to a higher degree in cultures with 58S sol-gel glasses. Northern blot analysis of the expression of osteoblastic markers revealed that osteoblasts retained their phenotype in both types of cultures. Interestingly, stimulation of alkaline phosphatase, bone sialoprotein, and osteocalcin was noticed at day 18 in sol-gel cultures when compared with that in control. These data confirm that 58S bioactive glasses are capable of supporting the growth and maturation of primary mouse osteoblasts. In addition, it was shown that 58S glasses affected the gene-expression profile, causing an up-regulation of the major bone markers. These results indicated that 58S sol-gel glasses appeared as suitable candidates for osteoblast scaffolds in the field of bone tissue engineering. PMID- 16345099 TI - Anti-inflammatory properties of micropatterned titanium coatings. AB - Prolonged inflammation and reactive oxygen species (ROS) generated around an implanted biosensor are the primary causes of the foreign body response, including encapsulation of biosensor membranes. We have previously demonstrated that TiO2 surfaces reduce ROS. Here we investigated the potential of using the anti-inflammatory properties of TiO2 in the design of biosensor membranes with improved long-term in vivo transport properties. Micropatterned Ti films were sputtered onto quartz surfaces in a series of hexagonally distributed dots with identical coverage area of 23% and dot size ranging from 5 to 100 microm. The antioxidant effect of the surfaces was investigated using a cell-free peroxynitrite donor assay and assays of superoxide released from stimulated surface-adhering neutrophils and macrophages. In all three assays, the amount of ROS was monitored using luminol-amplified chemiluminescence. Patterned surfaces in all experimental models significantly decreased ROS compared to the etched surfaces. In the cell-free experiment, the ROS reduction was only dependent on fractional surface coverage. In the cell experiments, however, a dot-size dependent ROS reduction was seen, with the largest reduction at the smallest dot size surfaces. These results indicate that micropatterned surfaces with small dots covering only 23% of the surface area exhibit similar antioxidative effect as fully covered surfaces. PMID- 16345100 TI - Characterization of human islet-like structures generated from pancreatic precursor cells in culture. AB - This study addresses the characterization of human islet-like structures generated from a newly discovered sparse population of precursor cells (Petropavlovskaia and Rosenberg, 2002) in the human pancreas. These cells may be progenitor cells capable of producing pancreatic cells suitable for the treatment of type 1 diabetes. The cells were cultured successfully in non-adherent stationary cultures and yielded, as an important first step, a 1.9-fold expansion in a serum-free medium developed specifically for this cell type. This expanded population grew as pancreatic cell aggregates, which were analyzed for islet-like characteristics. Specifically, through RT-PCR analyses and functionality assays, we show that cells within the population expressed all four of the endocrine hormone genes and proteins (insulin, glucagon, somatostatin, pancreatic polypeptide). As well, the expanded pancreatic precursor cell population exhibited glucose responsiveness although the produced cells appeared to be still primitive in nature. PMID- 16345102 TI - Robust estimation in mixed linear models with non-monotone missingness. AB - We introduce a model to account for abrupt changes among repeated measures with non-monotone missingness. Development of likelihood inferences for such models is hard because it involves intractable integration to obtain the marginal likelihood. We use hierarchical likelihood to overcome such difficulty. Abrupt changes among repeated measures can be well described by introducing random effects in the dispersion. A simulation study shows that the resulting estimator is efficient, robust against misspecification of fatness of tails. For illustration we use a schizophrenic behaviour data presented by Rubin and Wu. PMID- 16345101 TI - In vivo electrogene transfer of interleukin-12 inhibits tumor growth and lymph node and lung metastases in mouse mammary carcinomas. AB - BACKGROUND: Human breast cancer metastasizes mainly to lymph nodes, lungs, liver, and bone; in the majority of cases, it is the development of metastases which leads to death. In order to suppress mammary cancer metastasis, we applied in vivo electrogene transfer (non-viral method) as a means of interleukin-12 (IL-12) gene therapy on highly metastatic murine mammary cancer model. METHODS: Metastatic mammary tumors induced by inoculation in BALB/c female mice were treated by intratumoral injections of either a plasmid vector containing IL-12 or empty vector and then subjected to in vivo electrogene transfer once a week for 8 weeks. RESULTS: Treatment with IL-12 resulted in elevation of both IL-12 and IFNgamma levels in mammary tumors and in serum and intratumoral levels of CD4 and CD8 proteins were also increased. Tumor volumes and lymphatic and pulmonary metastases were significantly reduced. The histopathological changes induced by IL-12 characteristically included marked inflammation, increased apoptosis, decreased DNA synthesis, peripheral influx of significantly greater numbers of active macrophages, and reduced blood microvessel density, and apoptotic vascular endothelial cells were frequently seen. Western blotting showed decreases in VEGFR-3 of tumors exposed to IL-12 gene therapy. In adjuvant immunofluorescence studies, the CD31-positive endothelial cells of microvessels showed decreased VEGFR-3 expression in IL-12-treated tumors. However, apparent alterations in VEGFR-3 expression of podoplanin-positive lymphatic endothelial cells were not observed in IL-12-treated tumors. Although recombinant IL-12 did not inhibit tubular formation of human umbilical vein endothelial cells in a Matrigel assay, recombinant IFNgamma did completely suppress the tubular formation. CONCLUSIONS: In vivo electrogene transfer of IL-12 exerts strong anti-tumorigenic and anti metastatic effects likely due to T-cell-mediated immune responses as well as anti angiogenic action. PMID- 16345103 TI - Efficient estimation of log-normal means with application to pharmacokinetic data. AB - In this paper, the problem of interest is efficient estimation of log-normal means. Several existing estimators are reviewed first, including the sample mean, the maximum likelihood estimator, the uniformly minimum variance unbiased estimator and a conditional minimal mean squared error estimator. A new estimator is then proposed, and we show that it improves over the existing estimators in terms of squared error risk. The improvement is more significant with small sample sizes and large coefficient of variations, which is common in clinical pharmacokinetic (PK) studies. In addition, the new estimator is very easy to implement, and provides us with a simple alternative to summarize PK data, which are usually modelled by log-normal distributions. We also propose a parametric bootstrap confidence interval for log-normal means around the new estimator and illustrate its nice coverage property with a simulation study. Our estimator is compared with the existing ones via theoretical calculations and applications to real PK studies. PMID- 16345105 TI - Nonmechanical protein can have significant mechanical stability. PMID- 16345104 TI - Reversible cis-trans isomerization of a single azobenzene molecule. PMID- 16345106 TI - A copper(II)-catalyzed aza-Friedel-Crafts reaction of N-(2-pyridyl)sulfonyl aldimines: synthesis of unsymmetrical diaryl amines and triaryl methanes. PMID- 16345107 TI - Carbon nanotubes as intracellular transporters for proteins and DNA: an investigation of the uptake mechanism and pathway. PMID- 16345108 TI - Experimental and theoretical characterization of superoxide complexes [W2O6(O2-)] and [W3O9(O2-)]: models for the interaction of O2 with reduced W Sites on tungsten oxide surfaces. PMID- 16345109 TI - (13)C/(12)C isotope labelling to study leaf carbon respiration and allocation in twigs of field-grown beech trees. AB - In situ (13)C/(12)C isotopic labelling was conducted in field-grown beech (Fagus sylvatica) twigs to study carbon respiration and allocation. This was achieved with a portable gas-exchange open system coupled to an external chamber. This method allowed us to subject leafy twigs to CO(2) with a constant carbon isotope composition (delta(13)C of -51.2 per thousand) in an open system in the field. The labelling was done during the whole light period at two different dates (in June 2002 and October 2003). The delta(13)C values of respiratory metabolites and CO(2) that is subsequently respired during the night were measured. It was found that night-respired CO(2) is not completely labelled (only ca. 58% and 27% of new carbon is found in respired CO(2) immediately after the labelling in June 2002 and October 2003, respectively) and the labelling level progressively disappeared during the next day. It is concluded that the carbon respired by beech leaves after illumination was supplied by a mixture of carbon sources in which current carbohydrates were not the only contributors. In addition, as has been found in herbaceous plants, isotopic data before labelling showed that carbon isotope discrimination favoring the (13)C isotope occurred during the night respiration of beech leaves. PMID- 16345110 TI - Determination of M+4 stable isotope labeled cortisone and cortisol in human plasma by microElution solid-phase extraction and liquid chromatography/tandem mass spectrometry. AB - A sensitive microElution solid-phase extraction (SPE) liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of M+4 stable isotope labeled cortisone and cortisol in human plasma. In this method, M+4 cortisone and M+4 cortisol were extracted from 0.3 mL of human plasma samples using a Waters Oasis HLB 96-well microElution SPE plate using 70 microL methanol as the elution solvent, and chromatographed on a Waters Symmetry C18 column (4.6 x 50 mm, 3.5 microm). M+9 cortisone and M+9 cortisol were used as the internal standards. A PE Sciex API 4000 tandem mass spectrometer interfaced with the liquid chromatograph via a turboionspray source was used for mass analysis and detection. The selected reaction monitoring (SRM) of precursor --> product ion transitions were monitored at m/z 365.2 [M+H](+) --> 167.0 and at m/z 367.3 [M+H](+) --> 125.1 for M+4 cortisone and M+4 cortisol, respectively. The lower limit of quantitation was 0.1 ng mL(-1) and the linear calibration range was from 0.1 to 100 ng mL(-1) for both analytes. This method demonstrated to be very reproducible and reliable. PMID- 16345111 TI - Influence of serine O-glycosylation or O-phosphorylation close to the vJun nuclear localisation sequence on nuclear import. AB - Nuclear import triggered by the nuclear-localisation sequence (NLS) of the viral Jun (vJun) protein is mediated by phosphorylation of a serine close to the NLS. Since phosphorylation and glycosylation of serine residues are often in a reciprocal "yin-yang" relationship, we investigated whether glycosylation of this serine with O-linked N-acetylglucosamine (O-GlcNAc) would also regulate nuclear import via the vJun NLS. Peptides containing the vJun NLS with an adjacent O phosphorylated, O-GlcNAc-functionalised or unmodified serine, and equipped with an N-terminal biotin or a 7-nitrobenz-2-oxa-1,3-diazolyl (NBD) fluorescent label, were synthesised on the solid phase by means of an Fmoc/Boc strategy and a Pd0 sensitive HYCRON linker. Fluorescence-polarisation measurements on the NBD labelled peptides indicated that modification with phosphate or O-GlcNAc leads to a decrease in affinity to the import-mediating adapter protein, importin alpha, of about one order of magnitude compared to the unmodified NLS. Microinjection of biotinylated NLS peptide conjugated with fluorescently labelled avidin into NIH/3T3 and MDCK cells, revealed that avidin-unmodified-NLS peptide was rapidly imported into the nucleus. However, either phosphate or O-GlcNAc next to the NLS caused almost complete exclusion of the protein conjugate from nuclear import. These findings indicate that nuclear import by the vJun NLS might not be regulated by a "yin-yang" modification of an adjacent serine with phosphate or O GlcNAc. Rather, negative regulation of binding between the polybasic NLS and importin by a negatively charged or a bulky, uncharged residue appears likely. PMID- 16345112 TI - The RNA-cleaving bipartite DNAzyme is a distinctive metalloenzyme. AB - Much interest has focused on the mechanisms of the five naturally occurring self cleaving ribozymes, which, in spite of catalyzing the same reaction, adopt divergent strategies. These ribozymes, with the exception of the recently described glmS ribozyme, do not absolutely require divalent metal ions for their catalytic chemistries in vitro. A mechanistic investigation of an in vitro selected, RNA-cleaving DNA enzyme, the bipartite, which catalyzes the same chemistry as the five natural self-cleaving ribozymes, found a mechanism of significant complexity. The DNAzyme showed a bell-shaped pH profile. A dissection of metal usage indicated the involvement of two catalytically relevant magnesium ions for optimal activity. The DNAzyme was able to utilize manganese(II) as well as magnesium; however, with manganese it appeared to function complexed to either one or two of those cations. Titration with hexaamminecobalt(III) chloride inhibited the activity of the bipartite; this suggests that it is a metalloenzyme that utilizes metal hydroxide as a general base for activation of its nucleophile. Overall, the bipartite DNAzyme appeared to be kinetically distinct not only from the self-cleaving ribozymes but also from other in vitro-selected, RNA-cleaving deoxyribozymes, such as the 8-17, 10-23, and 614. PMID- 16345113 TI - Total synthesis and biological assessment of benzimidazole-based analogues of epothilone A: ambivalent effects on cancer cell growth inhibition. PMID- 16345114 TI - Micelles in ionic liquids: aggregation behavior of alkyl poly(ethyleneglycol) ethers in 1-butyl-3-methyl-imidazolium type ionic liquids. PMID- 16345115 TI - The use of 1H NMR microscopy to study proton-exchange membrane fuel cells. AB - To understand proton-exchange membrane fuel cells (PEMFCs) better, researchers have used several techniques to visualize their internal operation. This Concept outlines the advantages of using 1H NMR microscopy, that is, magnetic resonance imaging, to monitor the distribution of water in a working PEMFC. We describe what a PEMFC is, how it operates, and why monitoring water distribution in a fuel cell is important. We will focus on our experience in constructing PEMFCs, and demonstrate how 1H NMR microscopy is used to observe the water distribution throughout an operating hydrogen PEMFC. Research in this area is briefly reviewed, followed by some comments regarding challenges and anticipated future developments. PMID- 16345116 TI - Adsorbate clustering and proton transfer in zeolites: NMR spectroscopy and theory. PMID- 16345117 TI - Does silica surface catalyse peptide bond formation? New insights from first principles calculations. AB - The role that silica surface could have played in prebiotic chemistry as a catalyst for peptide bond formation has been addressed at the B3LYP/6-31+G(d,p) level for a model reaction involving glycine and ammonia on a silica cluster mimicking an isolated terminal silanol group present at the silica surface. Hydrogen-bond complexation between glycine and the silanol is followed by the formation of the mixed surface anhydride Si(surf)-O-C(=O)-R, which has been suggested in the literature to activate the C=O bond towards nucleophilic attack by a second glycine molecule, here simulated by the simpler NH3 molecule. However, B3LYP/6-31+G(d,p) calculations show that formation of the surface mixed anhydride Si(surf)-O-C(=O)-R is disfavoured (delta(r)G298 approximately 6 kcal mol(-1)), and that the surface bond only moderately lowers the free-energy barrier of the nucleophilic attack responsible for peptide bond formation (deltaG298(double dagger) approximately 48 kcal mol(-1)) in comparison with the uncatalysed reaction (deltaG298(double dagger) approximately 52 kcal mol(-1)). A further decrease of the free-energy barrier of peptide bond formation (deltaG298(double dagger) approximately 41 kcal mol(-1)) is achieved by a single water molecule close to the reaction centre acting as a proton-transfer helper in the activated complex. A possible role of strained silica surface defects on the formation of the surface mixed anhydride Si(surf)-O-C(=O)-R has also been addressed. PMID- 16345118 TI - Single-molecule conductance measurements of single- and double-stranded DNA oligonucleotides. PMID- 16345119 TI - Multilevel logistic regression modelling with correlated random effects: application to the Smoking Cessation for Youth study. AB - A multilevel logistic regression model is presented for the analysis of clustered and repeated binary response data. At the subject level, serial dependence is expected between repeated measures recorded on the same individual. At the cluster level, correlations of observations within the same subgroup are present due to the inherent hierarchical setting. Two random components are therefore incorporated explicitly within the linear predictor to account for the simultaneous heterogeneity and autoregressive structure. Application to analyse a set of longitudinal data from an adolescent smoking cessation intervention that motivated this study is illustrated. PMID- 16345120 TI - Development of two-dimensional gas chromatography/isotope ratio mass spectrometry for the stable carbon isotopic analysis of C(2)-C(5) non-methane hydrocarbons emitted from biomass burning. AB - A two-dimensional gas chromatography/combustion/isotope ratio mass spectrometry (2D-GC/C/IRMS) system was developed for stable carbon isotopic measurements of C(2)-C(5) non-methane hydrocarbons (NMHCs) in biomass burning smoke. The 2D GC/C/IRMS system successfully improved the accuracy and precision for the measurements of C(4) and C(5) saturated compounds in a smoke sample by selective injection of target compounds into a combustion furnace and consequently allowed us to provide complete baseline separation for all individual NMHCs. The analytical precision of the delta(13)C of each compound was better than 0.5 per thousand for more than 500 pmolC injections and 2.1 per thousand for 30 pmolC injections, which was estimated from replicate analysis of standard gases. This system was applied to the analysis of NMHCs in smoke samples collected from laboratory biomass burning experiments. From the combustion of three fuel materials (rice straw, pine wood, and maize), we found that the isotopic fractionation between fuel material and individual NMHCs is almost independent of the fuel material and thus the delta(13)C values of the fuel materials are reflected in delta(13)C values of most of NMHCs. However, only i-butane emitted from maize combustion showed anomalous (13)C-depletion of -11.6 per thousand relative to the delta(13)C value of maize. Such a large (13)C depletion suggests the specific isotopic fractionation process which is attributed to the maize combustion itself or the chemical properties of i-butane during production from a radical recombination reaction. PMID- 16345121 TI - Development of a liquid chromatography/tandem mass spectrometry assay for the quantification of lisinopril in human plasma. AB - A simple and sensitive liquid chromatography/tandem mass spectrometry method employing electrospray ionization, to quantify lisinopril in human plasma using pseudoephedrine hydrochloride as the internal standard (IS), has been developed and validated. A mixture of methanol and 0.1% formic acid in water (50:50, v/v) was used as the isocratic mobile phase. A simple liquid-liquid extraction procedure was used as sample preparation method. The method validation demonstrated the specificity, lower limit of quantification, accuracy, and precision of measurements. Selected reaction monitoring was specific for lisinopril and pseudoephedrine hydrochloride; no endogenous materials from blank plasma interfered with the analysis of lisinopril or the IS. The assay was linear over the concentration range 0.78-100 ng/mL. The correlation coefficients for the calibration curves ranged from 0.9984-0.9998. The intra- and inter-day precision, determined for quality control samples, were less than 4.18%. The method was employed in a pharmacokinetic study after oral administration of 10 mg lisinopril to 20 healthy volunteers. PMID- 16345122 TI - Fragmentation of mycosporine-like amino acids by hydrogen/deuterium exchange and electrospray ionisation tandem mass spectrometry. AB - The determination and identification of mycosporine-like amino acids (MAAs) from algae remain a major challenge due to the low concentration. Mass spectrometry (MS) can make an invaluable contribution in the search and identification of MAAs because of its high sensitivity, possibility of coupling with liquid chromatography, and the availability of powerful tandem mass spectrometric techniques. However, the unequivocal determination of the presence and location of important functional groups present on the basic skeleton of the MAAs is often elusive due to their inherent instability under MS conditions. In this study, the use of hydrogen/deuterium (H/D) exchange and electrospray ionisation tandem mass spectrometry (ESI-MS/MS) for characterisation of four MAAs (palythine, asterina, palythinol and shinorine) isolated from the macroalgae Gracilaria tenuistipitata Chang et Xia was investigated. The accurate-mass confirmation of the protonated molecules was performed on a Q-TOF instrument. We demonstrate that employing deuterium labelling in ESI-MS/MS analysis provides a convenient tool for the determination of new MAAs. Although the fragmentation patterns of MAAs were discussed earlier, to our knowledge, this is the first time that mechanisms are proposed. PMID- 16345123 TI - Systematic identification of N-acetylheparosan oligosaccharides by tandem mass spectrometric fragmentation. AB - Recently, a useful procedure for the preparation of both even- and odd-numbered series of N-acetylheparosan (NAH) oligosaccharides was established. The present report describes findings when these NAH oligosaccharides were subjected to comparative mass spectrometry (MS)/MS fragmentation analysis by matrix-assisted laser desorption/ionization (MALDI)-LIFT-time-of-flight (TOF)/TOF-MS/MS, and electrospray ionization (ESI) collision-induced dissociation (CID) MS/MS. The resultant fragment ions were systematically assigned to elucidate fragmentation characteristics. In the MALDI-LIFT-MS/MS experiments, all the NAH oligosaccharides underwent unique glycosidic cleavages that included B-Y ion cleavages (nomenclature system of Domon and Costello, Glycoconjugate J. 1988; 5: 397) at the C-1 side, and C-Z ion cleavages at the C-4 side, with respect to glucuronic acid (GlcA). In addition, (0,2)A and/or (0,2)X cross-ring cleavages were observed for relatively small oligosaccharides. The former observation clearly reflects the occurrence of a GlcA-N-acetylglucosamine (GlcNAc) alternating structure of NAH, while the latter feature implies the occurrence of the -beta-1-4-glucuronide linkage. Extensive glycosidic cleavages were also observed in the ESI-CID-MS/MS fragmentation, though cleavage specificity was less evident than in the case of MALDI-LIFT-TOF/TOF-MS/MS. The information obtained in this study should be valuable for understanding both biosynthetic and degradation processes of NAH and its derivatives including heparin and heparan sulfate, as well as artificially modified NAH oligosaccharides. PMID- 16345124 TI - New confidence intervals for the difference between two sensitivities at a fixed level of specificity. AB - For two continuous-scale diagnostic tests, it is of interest to compare their sensitivities at a predetermined level of specificity. In this paper, we propose three new intervals for the difference between two sensitivities at a fixed level of specificity. These intervals are easy to compute. We also conduct simulation studies to compare the relative performance of the new intervals with the existing normal- approximation-based interval proposed by Wieand et al. Our simulation results show that the newly proposed intervals perform better than the existing normal-approximation-based interval in terms of coverage accuracy and interval length. PMID- 16345125 TI - Investigation of the profiling depth in matrix-assisted laser desorption/ionization imaging mass spectrometry. AB - Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry is generally considered to be a surface analysis technique. In this report, the profiling depth of imaging mass spectrometry was examined. MALDI matrix solution was found to be able to gain access to the tissue interior and extract analyte molecules to the tissue surface. As a consequence, prazosin, a small molecule pharmaceutical compound, located as deep as 40 microm away from the surface was readily detected after matrix application. Likewise, cytochrome c, a 12 kDa protein, was also detectable from the tissue interior. Moreover, for prazosin, not only the extent of matrix effect, but also the extraction efficiency of the matrix solvent appeared to be dependent on the type of tissue. These results indicated that experimental conditions that decrease the matrix solvent evaporation during matrix application may increase analyte extraction efficiency and hence sensitivity of the analysis. Furthermore, thin sections should be used to avoid differential extraction efficiency of matrix solvent in different tissues for whole-body analysis. PMID- 16345126 TI - The effect of tetrahydrofuran as solvent on matrix-assisted laser desorption/ionization and electrospray ionization mass spectra of functional polystyrenes. AB - It is demonstrated that tetrahydrofuran (THF) should be used with caution as a solvent in polymer sample preparation for matrix-assisted laser desorption/ionization (MALDI) analyses. The presence of peroxides in THF may cause the oxidation of chemically active groups. This effect is illustrated in the case of S-containing polystyrene derivatives. The oxidation of the trithiocarbonate group resulted in the formation of poly(styrene)sulfonic acids, R(CH(2)CHPh)(n)SO(3)H, which was detected in negative mode by MALDI and electrospray ionization (ESI) methods after the samples remained in THF for several hours. PMID- 16345127 TI - Detection of royal jelly adulteration using carbon and nitrogen stable isotope ratio analysis. AB - Stable isotope ratios ((13)C/(12)C and (15)N/(14)N) were measured in royal jelly (RJ) samples by isotope ratio mass spectrometry (IRMS) to evaluate authenticity and adulteration. Carbon and nitrogen isotope contents (given as delta values relative to a standard, delta(13)C, delta(15)N) of RJ samples from various European origins and samples from commercial sources were analyzed. Uniform delta(13)C values from -26.7 to -24.9 per thousand were observed for authentic RJ from European origins. Values of delta(15)N ranged from -1.1 to 5.8 per thousand depending on the plant sources of nectars and pollen. High delta(13)C values of several commercial RJ samples from -20.8 to -13.3 per thousand indicated adulteration with high fructose corn syrup (HFCS) as a sugar source. Use of biotechnologically produced yeast powder as protein source for the adulterated samples was assumed as delta(15)N values were lower, as described for C(4) or CAM plant sources. RJ samples from authentic and from adulterated production were distinguished. The rapid and reliable method is suitable for urgent actual requirements in food monitoring. PMID- 16345128 TI - Enantiomeric separation and quantification of pindolol in human plasma by chiral liquid chromatography/tandem mass spectrometry using staggered injection with a CTC Trio Valve system. AB - Pindolol is a non-selective beta-adrenergic antagonist (beta-blocker) for the treatment of cardiovascular diseases such as hypertension and angina pectoris. It has one chiral center, and, therefore, two optical isomers. It was essential to develop an enantioselective assay to measure each enantiomer in human plasma. However, separation of enantiomers using chiral chromatography usually requires relatively long retention times. This can pose a problem for rapid turnaround of a large number of samples (i.e., clinical studies). In the present study, a simple and sensitive chiral liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method was developed and validated for the determination of S-(-)- and R-(+)-pindolol in human plasma. To increase throughput, staggered sample injection was employed using a CTC Trio Valve system on a CTC HTS PAL autosampler. The method exhibited good intra- and inter-day accuracy and precision, and was linear over a dynamic range of 250 pg/mL to 250 ng/mL for each pindolol enantiomer. Intra- and inter-day accuracy ranged between 90.0-106% and 91.6-104% for both quality control (QC) samples of S-(-)- and R-(+) pindolol, respectively. The respective intra- and inter-day precision ranged between 4.24-7.86% and 4.98-10.4%. PMID- 16345129 TI - A preference-based measure for test performance with an application to prenatal diagnostics. AB - Clinical epidemiology generally uses the receiver operating characteristic curve to summarize the accuracy of a diagnostic test and to compare the relative performance of different tests. This paper extends this concept to include the utility gains and losses of true and false test outcomes over the range of a priori risk. A utility index is developed first in situations where test accuracy is exogenously given, second where the test cutpoint can be chosen by the clinician according to the patient's a priori risk and preferences. By integrating over the a priori risk range, we derive an overall measure for a test's performance weighted by utility gains and losses. An example in prenatal diagnostics finally illustrates the clinical uses of the novel approach. Integrating patients' preference into clinical decision making will lead to different cutpoints and different assessments of test performance compared to unweighted policies. PMID- 16345130 TI - Profiling of ecdysteroids in complex biological samples using liquid chromatography/ion trap mass spectrometry. AB - A sensitive method using high-performance liquid chromatography coupled to a mass spectrometer with electrospray ionization source (HPLC/ESI-MS) was developed for detection of ecdysteroids in biological samples. We report here for the first time that ecdysteroids can be classified into three groups based on ESI full-scan mass spectra: group 1 (ecdysone (E), 2-deoxyecdysone (2dE), 2,22-dideoxyecdysone (3beta5beta-KT), and 3alpha5alpha[H]-dihydroxycholest-7-en-6-one (3alpha5alpha KD)), in which loss of one molecule of water from the protonated molecular ion ([M+H](+)) represents the dominant ion; group 2 (20-hydroxyecdysone (20E), makisterone A (MakA), 3beta5beta-KD, and 3beta5alpha-KD), in which [M+H](+) is a major ion but some water loss is observed; and group 3 (muristerone A (MurA) and ponasterone A (PonA)), in which [M+H](+) is the dominant ion with no water loss observed. Based on the analytical procedure in combination with structural information from the group classification and with the application of source induced dissociation, we identified free ecdysteroids in biological samples: 20,26-dihydroxyecdysone and ecdysonic acid in the larval hemolymph, and the progressive metabolism of 26-hydroxyecdysone (26E) to 3alpha-26E from day-1 to day-3 embryos of the tobacco hornworm Manduca sexta. PMID- 16345131 TI - Tandem mass spectrometry of coprogen and deferoxamine hydroxamic siderophores. AB - Mechanisms of fragmentation of hydroxamic siderophores are proposed comparing deuterated and nondeuterated samples. Standard siderophores (e.g. deferoxamine and coprogen) were directly injected into both ion trap and linear quadrupole mass spectrometers with electrospray ionization (ESI). Four and two fragmentation steps were carried out for deferoxamine and coprogen (analyzed by positive and negative ESI, respectively). Deferoxamine cleavages occurred in both peptide and hydroxamic bonds while the coprogen fragmentation pattern is more elaborate, since it contains Fe(III) in its structure. PMID- 16345132 TI - Quantitative determination of piritramide in human plasma and urine by off- and on-line solid-phase extraction liquid chromatography coupled to tandem mass spectrometry. AB - A method for the liquid chromatography/tandem mass spectrometric (LC/MS/MS) quantification of piritramide, a synthetic opioid, in plasma after conventional off-line solid-phase extraction (SPE) and in urine by on-line SPE-LC/MS/MS in positive electrospray mode was developed and validated. Applicability of the on line approach for plasma samples was also tested. Deuterated piritramide served as internal standard. For the off-line SPE plasma method mixed cation-exchange SPE cartridges and a 150 x 2 mm C18 column with isocratic elution were used. For the on-line SPE method, a Waters Oasis HLB extraction column and the same C18 analytical column in a column-switching set-up with gradient elution were utilized. All assays were linear within a range of 0.5-100 ng/mL with a limit of detection of 0.05 ng/mL. The intra- and interday coefficients of variance ranged from 1.3 to 6.1% for plasma and 0.5 to 6.4% for urine, respectively. The extraction recovery for the off-line plasma assay was between 90.7 and 100.0%. Influence of matrix effects, and freeze/thaw and long-term stability were validated for both approaches; influence of urine pH additionally for quantification in urine. PMID- 16345133 TI - Experimental apparatus for investigation of sputtering and secondary ion emission induced by energetic ion beams. AB - The construction of an experimental apparatus, for investigation of implantation, secondary ion emission and sputtering processes, during irradiation of samples with an ion beam of up to 70 keV energy, is described. The basis of the apparatus is an electromagnetic mass separator equipped with a quadrupole mass spectrometer located in the collector chamber. The computer data acquisition control system makes it possible to perform the experimental measurements with high accuracy and precision. Preliminary results of secondary ion mass spectral measurements, obtained for C, Al, Si and Cu targets bombarded with Ar(+) and Kr(+) ions, are presented. PMID- 16345134 TI - Investigation of disulfonamide ligands derived from o-phenylenediamine and their Pb(II) complexes by electrospray ionization mass spectrometry. AB - An electrospray ionization mass spectrometry (ESI-MS) method, in both positive and negative ion modes, was developed for characterization of disulfonamide ligands derived from o-phenylenediamine and their Pb(II) complexes. For the ligands, negative ion mode ESI-MS in methanolic solutions gave simple and easily interpretable mass spectra. However, the spectra of Pb complexes were not readily interpretable under the same conditions. Protonated ligands and their Pb(II) complexes were observed in methanolic solutions by ESI-MS in positive ion mode. The formation of Na(+), K(+), or NH(4) (+) adducts was also observed, complicating the mass spectra and decreasing the signal intensity. In order to optimize the detection of the ligands and the Pb complexes, a method was developed by adding NaOAc in the solutions. The presence of 0.2 mM NaOAc simplified the mass spectra of the ligands and the Pb complexes, and significantly increased sensitivity in both negative and positive ion modes. This modification makes ESI-MS in both modes suitable for characterization of sulfonamide ligands and their Pb complexes, thus providing a potentially powerful tool for evaluating formation of metal complexes and screening combinatorial ligand libraries. PMID- 16345135 TI - The use of mass spectrometry to determine location and extent of N-glycosylation on folate binding protein from bovine milk. PMID- 16345136 TI - Detection of penicillin residues in bovine oral fluid (saliva) by liquid chromatography/tandem mass spectrometry. PMID- 16345137 TI - High-precision, automated stable isotope analysis of atmospheric methane and carbon dioxide using continuous-flow isotope-ratio mass spectrometry. AB - Small-scale developments have been made to an off-the-shelf continuous-flow gas chromatography/isotope-ratio mass spectrometry (CF-GC/IRMS) system to allow high precision isotopic analysis of methane (CH(4)) and carbon dioxide (CO(2)) at ambient concentrations. The repeatability (1sigma) obtainable with this system is 0.05 per thousand for delta(13)C of CH(4), 0.03 per thousand for delta(13)C of CO(2), and 0.05 per thousand for delta(18)O of CO(2) for ten consecutive analyses of a standard tank. An automated inlet system, which allows diurnal studies of CO(2) and CH(4) isotopes, is also described. The improved precision for CH(4) analysis was achieved with the use of a palladium powder on quartz wool catalyst in the combustion furnace, which increased the efficiency of oxidation of CH(4) to CO(2). The automated inlet further improved the precision for both CH(4) and CO(2) analysis by keeping the routine constant. The method described provides a fast turn-around in samples, with accurate, reproducible results, and would allow a long-term continuous record of CH(4) or CO(2) isotopes at a site to be made, providing information about changing sources of the gases both seasonally and interannually. PMID- 16345138 TI - Laser desorption/ionization time-of-flight mass spectrometry of squalene in oil samples. PMID- 16345139 TI - [Best disease]. PMID- 16345140 TI - [Sentinel lymph node biopsy]. PMID- 16345141 TI - Duchenne de Boulogne (1806-1875). PMID- 16345142 TI - [How to better inform pregnant women? Recommendations of the HAS for health professionals (April 2005)]. PMID- 16345143 TI - The efficacy of a fluoride chewing gum on salivary fluoride concentration and plaque pH in children. PMID- 16345144 TI - An FDA reviewer battles the drug his boss approved: private letter gets Dr. Misbin pulled from diabetes case but he pursues it anyway. PMID- 16345146 TI - Tiotropium effective in treatment of COPD. PMID- 16345147 TI - Distribution and significance of fecal indicator organisms in the upper chesapeake bay. AB - [This corrects the article on p. 626 in vol. 30.]. PMID- 16345148 TI - Metabolic activities of the intestinal microflora of a deep-sea invertebrate. AB - The intestinal microflora of deep-sea amphipods, in enrichment culture employing starch, urea, and N-acetyl-d-glucosamine and when examined under simulated in situ conditions, exhibited growth rates and substrate conversion approximately equal to, or greater than, atmospheric controls during short-term incubation. These observations are significant since these microorganisms may play an important role in biodegradation in the deep sea. PMID- 16345149 TI - Kinetics of growth and substrate uptake in a biological film system. AB - The rates of growth and substrate uptake in a biological film continuous-flow reactor were studied. The experiments were performed with high fluid velocities to bring the reactor operation to the reaction-controlled regime, thus avoiding external diffusional resistances. The glucose uptake experiments were performed with small film thicknesses so that full substrate penetration within the entire film thickness could be obtained. In this way, the catalyst effectiveness factor was 1.0 and the observed rate was the true, or intrinsic, rate. The results of the experiments indicate that both the intrinsic rate of substrate uptake and the rate of film growth are independent of the substrate concentration remaining in the reactor (zero-order reactions). However, the value of the initial substrate concentration when the film is in the early stages of growth defines the magnitude of both the rate of uptake and growth. This effect of the initial substrate concentration follows a saturation-function pattern. PMID- 16345150 TI - Improved specimen coating technique for scanning electron microscope observation of decomposer microorganisms. AB - Sputter coating of leaf litter microbe samples provides scanning electron microscope images with greater information content than either vacuum evaporation of thin metal coatings or tissue conductance. PMID- 16345152 TI - Kinetics of a bacterial culture growth: validity of the affinity rule in biological systems. AB - [This corrects the article on p. 1000 in vol. 30.]. PMID- 16345151 TI - Comparison of surface pore morphology of two brands of membrane filters. AB - The surface pore morphology of two brands of membrane filters was studied by using scanning electron microscopy. The differences observed are presented as a possible explanation for reported discrepancies in coliform recovery from water. PMID- 16345153 TI - New type of turbidostat with intermittent determination of cell density outside the culture vessel. AB - An inexpensive turbidostat featuring intermittent determination of optical density in an interchangeable measuring tube and incorporating electronic circuits for the automated mechanics is described. PMID- 16345154 TI - Growth requirements of san francisco sour dough yeasts and bakers' yeast. AB - The growth requirements of several yeasts isolated from San Francisco sour dough mother sponges were compared with those of bakers' yeast. The sour dough yeasts studied were one strain of Saccharomyces uvarum, one strain of S. inusitatus, and four strains of S. exiguus. S. inusitatus was the only yeast found to have an amino acid requirement, namely, methionine. All of the yeasts had an absolute requirement for pantothenic acid and a partial requirement for biotin. Inositol was stimulatory to all except bakers' yeast. All strains of S. exiguus required niacin and thiamine. Interestingly, S. inusitatus, the only yeast that required methionine, also needed folic acid. For optimal growth of S. exiguus in a molasses medium, supplementation with thiamine was required. PMID- 16345155 TI - Microbial studies on shelf life of cabbage and coleslaw. AB - The microbiology of a common commercial type of coleslaw was investigated with the objective of extending its shelf life at refrigerator temperature by delaying microbiological spoilage. Cabbage, its principal ingredient, had a total bacterial count of about 10/g. Microbial growth in cabbage was prevented by storage at 1 C but not at 10 C or above. In coleslaw, the cabbage flora died and was replaced by the flora of the cultured sour cream contained in the dressing. At 14 C, the total count increased and the coleslaw deteriorated organoleptically. At 7 C, bacterial growth was suppressed but organoleptic deterioration occurred as rapidly as at 14 C. Thus, the deterioration was caused primarily by the physiological breakdown of plant tissue rather than by microorganisms, as was the original premise. PMID- 16345157 TI - Chilling Cells Enhances the Bactericidal Action of Fatty Acids on Escherichia coli. AB - [This corrects the article on p. 153 in vol. 31.]. PMID- 16345156 TI - Delayed release nutrient supplement for mushroom culture. AB - The disadvantages associated with the supplementation of noncomposted nutrients to mushroom compost at spawning were largely overcome by encapsulating microdroplets of vegetable oil within a protein coat that was denatured with formaldehyde. Increases in mushroom yield of 60% were obtained. Delayed nutrient release was indicated by prolonged stimulation of yields beyond the first few flushes. PMID- 16345158 TI - Effect of short-chain Fatty acids extracted from beef cattle manure on germination and seedling development. AB - Composted and fresh beef cattle manure samples were extracted with distilled water, acetone, methanol, 2 N sodium hydroxide, 2 N hydrochloric acid, and ether. Bioassay techniques, using the extracts, showed that composted manure extracts had limited effect on seed germination and seeding development of wheat and sorghum. All the extracts of fresh manure, other than distilled water, retarded germination. Acetic, butyric, propionic, valeric, and isovaleric acids were found in ether extracts of fresh manure at average concentrations of 348, 876, 578, 34, and 19 mug/g, respectively, on a dry-weight basis. However, only trace amounts of these acids were present in composted manure. Propionic acid up to the 200-mug/ml level stimulated seedling growth. Acetic and butyric below the 200-mug/ml level had no detrimental or beneficial effects on seedling development. When acetic, butyric, and propionic acids were mixed in equal parts, germination and seedling growth were reduced at all levels (50 to 500 mug/ml). PMID- 16345159 TI - Microbial degradation of lignocellulose: the lignin component. AB - A new procedure was developed for the study of lignin biodegradation by pure or mixed cultures of microorganisms. Natural lignocelluloses were prepared containing C in primarily their lignin components by feeding plants l-[U C]phenylalanine through their cut stems. Lignin degradation was observed in numerous soils by monitoring evolution of CO(2) from [C]lignin-labeled oak (Quercus albus), maple (Acer rubrum), and cattail (Typha latifola). An organism (Thermonospora fusca ATCC 27730) that is known to degrade cellulose but not lignin was shown to grow on lignocellulose in the presence of [C]lignocelluloses without evolution of CO(2). A known lignin degrader (a white-rot fungus, Polyporus versicolor) was shown to readily evolve CO(2) from damp C-labeled cattail and C-labeled maple. PMID- 16345160 TI - Changes in Resident Bacteria, pH, Sucrose, and Invert Sugar Levels in Sugarbeet Roots During Storage. AB - Stored sugarbeet roots began fermenting within 24 h after oxygen was depleted at 26 C when the resident bacterial populations increased dramatically. Most of the bacteria present after anaerobic storage for 7 days at 26 C could hydrolyze sucrose in vitro. Although pH and sucrose levels decreased and invert sugar levels increased with time in aerobic storage at 26 C, these processes were significantly accelerated in beets stored anaerobically at 26 C. Under oxygen depleted storage conditions at 26 C, the sucrose content was almost completely lost after 21 days. PMID- 16345162 TI - In situ morphology of nitrifying-like bacteria in aquaculture systems. AB - [This corrects the article on p. 423 in vol. 31.]. PMID- 16345161 TI - N(2) Fixation Associated with Decaying Leaves of the Red Mangrove (Rhizophora mangle). AB - N(2) (C(2)H(2)) fixation was associated with decaying leaves of Rhizophora mangle. The process was predominantly anaerobic, with about two-thirds of the nitrogenase activity being light dependent. Average N(2) fixation rates in the light were 11 mug of N per g (dry weight) per h for leaves that had decayed for 2 to 3 weeks. This nitrogen input is probably significant in the estuarine, detrital food chains linked to R. mangle. PMID- 16345163 TI - Patchiness in the distribution of planktonic heterotrophic bacteria in lakes. AB - By using conventional dilution plating procedures, the heterotrophic planktonic bacteria in Lake Washington were found to be distributed in small-scale (<1.0 ml) and large-scale (>1 m in horizontal distance) patches. Recommendations are made on sampling and enumeration procedures to minimize the effect of patchiness on obtaining an accurate estimate of bacterial numbers. PMID- 16345164 TI - Influence of Organic Solvents on Chalcopyrite Oxidation Ability of Thiobacillus ferrooxidans. AB - It has been shown that organic solvents used primarily for the extraction of metals from aqueous leach liquors decrease both the surface tension of the aqueous phase and the chalcopyrite oxidation ability of Thiobacillus ferrooxidans. For the reagents and modifiers investigated, the order of inhibition was found to be LIX 70 < LIX 73 < LIX 71 < LIX 64N < LIX 65N < TBP approximately isodecanol approximately nonylphenol < LIX 63 <<< D(2)EHPA approximately Kelex 100 < Kelex 120 <<< Alamine 336 approximately Alamine 308 approximately Alamine 310 < Alamine 304 < Adogen 381 approximately Aliquat 336 < Adogen 364. To avoid limitation in bacterial activity, organic matter should be removed from the recycling liquor prior to leaching. PMID- 16345165 TI - Easily constructed soil percolation apparatus. AB - The construction of a soil percolator from a disposable Nalgene sterilizing filter is described. PMID- 16345166 TI - Requirement for a growth substrate during lignin decomposition by two wood rotting fungi. AB - Decomposition of C-labeled lignin to CO(2) by the lignin-decomposing fungi Phanerochaete chrysosporium and Coriolus versicolor required a growth substrate such as cellulose or glucose. Growth with lignin as sole carbon addition to an otherwise complete medium was negligible. PMID- 16345167 TI - Utilization of Lactose, Glucose, and Galactose by a Mixed Culture of Streptococcus thermophilus and Lactobacillus bulgaricus in Milk Treated with Lactase Enzyme. AB - The mechanism responsible for an increased rate of acid production when yogurt starter cultures are grown in milk treated with lactase enzyme was investigated by studying carbohydrate utilization and acid development by a pure culture of Streptococcus thermophilus and a mixed yogurt starter culture consisting of S. thermophilus and Lactobacillus bulgaricus. In milk containing glucose, galactose, and lactose, glucose and lactose (but not free galactose) were fermented. Fermentation of lactose in control milk was accompanied by the release of free galactose, with the result that carbohydrate utilization was less efficient than in treated milk. This phenomenon also occurred when lactose was fermented by S. thermophilus in broth culture. Carbohydrate utilization by the mixed yogurt culture was more rapid when the lactose in milk was partially prehydrolyzed. Our results suggest that the more rapid acid development that took place when a mixed yogurt starter culture was grown in milk containing prehydrolyzed lactose was the result of a more rapid and efficient utilization of carbohydrate by S. thermophilus when free glucose in addition to lactose was available for fermentation. The evidence presented also suggests that uptake and utilization of glucose and lactose by S. thermophilus are different in broth and milk cultures. PMID- 16345168 TI - Acetylene reduction (nitrogen fixation) by enterobacteriaceae isolated from paper mill process waters. AB - Using selective media containing galactitol, over 130 Enterobacteriaceae have been isolated from paper mill process waters collected from different localities. These bacteria were extensively characterized and tested for acetylene-reducing (nitrogen-fixing) activity under anaerobic conditions. High activity was found in representatives of Klebsiella pneumoniae, Enterobacter aerogenes, Enterobacter cloacae, Erwinia herbicola, Citrobacter freundii, Citrobacter intermedius, and Escherichia coli. Under argon, nitrogenase synthesis was generally not repressed by 5 mM l-glutamate, l-aspartate, l-leucine or Casamino Acids (0.5 g/liter). In many strains, both the specific activities (nanomoles of C(2)H(4) per minute per milligram of protein) and the activities (nanomoles of C(2)H(4) per minute) had considerably declined after 24 h. In three selected strains, activity in intact cells grown under nitrogen was unaffected by the presence during assay of 10 mM l amino acids or ammonium acetate. All of the strains examined were tolerant towards inactivation of nitrogen-fixing activity by 1.8% (vol/vol) oxygen during assay, and inactivation by up to 10% oxygen was partly reversible. Representatives of the six taxa synthesized nitrogenase in stirred aerobic cultures, though the protein concentrations attained were lower than under anaerobic conditions. It seems reasonable to suggest that under natural conditions, nitrogen fixation is able to contribute significantly to the nitrogen economy of the cells. PMID- 16345169 TI - Bacteria in the air of housed Swine units. AB - Two housed, swine-growing-finishing units were studied for numbers of total bacteria, fecal coliforms, Staphylococcus, and Salmonella in the air. At 30.5 and 122 cm from the floor, total colony-forming particles, as tested, averaged 3.4 x 10 and 1.3 x 10/m of air, respectively; whereas fecal coliform counts averaged 24 x 10 and 1.9 x 10/m of air, respectively. Only 41% of the organisms growing on Staphylococcus 110 medium tested as Staphylococcus. Of 458 Staphylococcus isolates, 5 were coagulase-positive. No Salmonella were detected in the air of the units tested. PMID- 16345170 TI - Penetration of Rhizopus oligosporus into Soybeans in Tempeh. AB - Histological observations were made on the penetration of hyphae of Rhizopus oligosporus into soybean cotyledons in tempeh, an Indonesian soybean food. Hyphal penetrations averaged one per 1,400 mum (+/-390 mum) on the curved (outer) cotyledon surface and one per 1,010 mum (+/-340 mum) on the flat (inner) one. Hyphae infiltrated to a depth of 742 mum, or about 25% of the average width of a soybean cotyledon. This previously unreported degree of penetration offers partial explanation for the rapid physical and chemical changes in soybeans during tempeh fermentation. PMID- 16345171 TI - High Production of Kojic Acid Crystals by Aspergillus parasiticus UNBF A12 in Liquid Medium. AB - The crystalline compound produced in large quantity in liquid medium by Aspergillus parasiticus UNBF A12, a high aflatoxin-producing strain isolated from the air in the Federal District of Brazil, was identified as kojic acid. The effect of pH on the production of crystalline kojic acid and aflatoxins by the strain was studied. Fourteen single spore isolates were evaluated for their capacity to produce kojic acid crystals and aflatoxins. PMID- 16345172 TI - Viscous product from activated sludge by methanol fermentation. AB - Aeration of activated sludge with 3 to 4% added methanol for 5 to 7 days yields an odorless, highly viscous (5,000 to 10,000 centipoise), black, pudding-like product containing glycan(s) linked other than alpha-1-4 or beta-1-3. Backseeding gives maximum thickening in 3 to 4 days. Incomplete acid hydrolysis of the black product gives a 0.27% solution of reducing sugars (75% glucose) which is an 11.4% yield from the added methanol. Backseeding into either centrifuge supernatant or 0.1% yeast extract in tap water gives a light-colored polymer. Viscosity decreases during extended sterile cold storage. A 5% salt addition lowers viscosity one-half. From 6 to 12 colony types appear on plating backseeded media, but none of these isolates is a reliable polymer former. PMID- 16345173 TI - Stimulatory Effect of Malo-Lactic Fermentation on the Growth Rate of Leuconostoc oenos. AB - Although l-malic acid is not an energy source for the malo-lactic organism Leuconostoc oenos (L. citrovorum) ML 34, the growth rate of the organism was found to be greatly increased by malo-lactic fermentation (the decarboxylation of l-malic acid to l-lactic acid). The stimulation was especially striking at the low pH (below pH 4) of wine, the natural habitat of this bacterium. The stimulation of growth did not result from changes in pH that accompany malo lactic fermentation. Thus, these results suggest a biological function of malo lactic fermentation. PMID- 16345174 TI - Regulation of lactose fermentation in group N streptococci. AB - Group N streptococci, which have the lactose phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) and phospho-beta-d-galactosidase (beta-Pgal), grew rapidly on lactose and converted more than 90% of the sugar to l-lactate. In contrast, Streptococcus lactis 7962, which does not have a beta-Pgal, grew slowly on lactose and converted only 15% of the sugar to l-lactate. With glucose and galactose, this strain had growth rates and fermentation patterns similar to those of other S. lactis strains, suggesting that the rapid and homolactic fermentation of lactose that is characteristic of group N streptococci is dependent upon a functional PEP-dependent PTS and the presence of beta-Pgal. Seventeen strains of group N streptococci were examined for the activator specificities of pyruvate kinase and lactate dehydrogenase. The properties of each enzyme from all the strains, including S. lactis 7962, were similar. Pyruvate kinase had a broad activator specificity, whereas activation of lactate dehydrogenase was specific for ketohexose diphosphate. All intermediates of lactose metabolism from the hexose phosphates to the triose phosphates activated pyruvate kinase. No activation was obtained with adenosine 5'-monophosphate. K and Mg were required for pyruvate kinase activity but could be replaced by NH(4) and Mn, respectively. Lactate dehydrogenase was activated equally by fructose-1,6 diphosphate and tagatose-1,6-diphosphate, the activation characteristics being pH dependent. The roles of pyruvate kinase and lactate dehydrogenase in the regulation of lactose fermentation by group N streptococci are discussed. PMID- 16345175 TI - Preparation of Agar-Agar from the Red Seaweed Pterocladia capillacea off the Coast of Alexandria, Egypt. AB - The effect of different treatments on the quality of agar produced from Pterocladia has been studied, and the conditions for the production of material of good quality have been standardized. In the modified process, sun-bleached seaweed was washed well in water, soaked for 24 h, and then ground to a pulp and rinsed again in water. The pulp was then extracted with water (weed-to-water ratio, 1:30) under pressure for 2 h after adjusting the pH to 6 by the addition of acetic acid. The agar gel, after freeze thawing, was bleached with NaClO before drying in a current of hot air. Pretreatment of the seaweed with alkali at 80 degrees C for 2 h prior to extraction was found to improve the quality of agar to a very great extent. PMID- 16345176 TI - Paint Deterioration as a Result of the Growth of Aureobasidium pullulans on Wood. AB - Growth of Aureobasidium was noted on both painted and unpainted wood surfaces but not on painted glass slides, indicating that the source of carbon and energy for growth was not associated with paint components but was probably supplied by the wood. Several aromatic compounds related to aromatic extractives of wood were shown to support growth of this fungus. PMID- 16345177 TI - Enhanced dissolution of oil shale by bioleaching with thiobacilli. AB - Oil shale was subjected to bioleaching by cultures of thiobacilli. From X-ray, electron microprobe, and thin-section petrographic analysis, the shale matrix was found to contain tightly bonded carbonate minerals. When subjected to the bioproduced acids, these carbonate minerals were removed successively from the shale matrix. This process created pits and cavities which were gradually enlarged as indicated by scanning electron micrographs of samples subjected to leaching for varying lengths of time. At the end of 14 days, essentially all available carbonates had been depleted from the solid matrix. The effected increase in porosity and permeability of the oil shale then enhanced the exposure of fuel precursors, thus facilitating their production and conversion. PMID- 16345178 TI - Myxobacters from arid mexican soil. AB - Myxobacters were found to be common inhabitants of the arid soils from the Monterrey, Nuevo Leon, Mexico, area. Thirteen species of the genera Myxococcus, Archangium, Cystobacter, Stigmatella, Polyangium, and Chondromyces were isolated on a mineral salts agar supplemented with bakers' yeast and filter paper. Greater species diversity per soil sample was found in the region receiving 400 to 800 mm of annual rainfall as compared with soils from an area having only 200 to 400 mm of rainfall. PMID- 16345179 TI - Alcohol dehydrogenase activities of wine yeasts in relation to higher alcohol formation. AB - Alcohol dehydrogenase activities were examined in cell-free extracts of 10 representative wine yeast strains having various productivities of higher alcohols (fusel oil). The amount of fusel alcohols (n-propanol, isobutanol, active pentanol, and isopentanol) produced by the different yeasts and the specific alcohol dehydrogenase activities with the corresponding alcohols as substrates were found to be significantly related. No such relationship was found for ethanol. The amounts of higher alcohols formed during vinification could be predicted from the specific activities of the alcohol dehydrogenases with high accuracy. The results suggest a close relationship between the control of the activities of alcohol dehydrogenase and the formation of fusel oil alcohols. Also, new procedures for the prediction of higher alcohol formation during alcoholic beverage fermentation are suggested. PMID- 16345180 TI - Definition of bacteriophage groups according to their lytic action on mesophilic lactic streptococci. AB - The lytic activity of 132 phages isolated during slow acid production in cheese factories situated in all the dairying regions of France during the past 16 years has been determined on 291 strains of mesophilic lactic streptococci. The results have been treated according to a method of analysis of data so as to establish a classification. Six groups of phages have thus been formed. Sixty-six percent of the phages studied, which are very similar and for the most part nonspecific to one species, have been gathered together in one group. On the other hand, a classification of the bacterial strains has been made on their sensitivity to the phages. Six groups, each corresponding to one of these groups of phages, have thus been defined. One of them accounts for 40% of the strains studied, of which certain ones are sensitive to a large number of phages. PMID- 16345181 TI - Products Formed from Analogues of 1,1,1-Trichloro-2,2-Bis(p-Chlorophenyl) Ethane (DDT) Metabolites by Pseudomonas putida. AB - Cultures of Pseudomonas putida growing in solutions with diphenylmethane as sole carbon source formed 1,1,1',1'-tetraphenyldimethyl ether. The product was identified by gas chromatography, mass spectrometry, and infrared and nuclear magnetic resonance spectrometry. The formation of benzophenone, benzhydrol, and phenylglycolic acid was established by gas chromatography and mass spectrometry. Similar techniques also revealed that phenylacetic acid was a major metabolite. Resting cell suspensions converted benzhydrol to phenyl-glycolic acid and products tentatively identified as hydroxybenzhydrols and a hydroxybenzophenone. Cell suspensions of the bacterium also converted the tetraphenyldimethyl ether to benzhydrol and benzophenone. Possible pathways for the degradation of these analogues of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) metabolites are discussed. PMID- 16345182 TI - Aeromonas distribution and survival in a thermally altered lake. AB - Par Pond is a thermally enriched monomictic southeastern lake which receives heated effluent from a production nuclear reactor. Fish populations in the lake have lesions of epizooty from which Aeromonas spp. are readily isolated. Distribution and population densities of Aeromonas in the water column were measured along an oxygen and temperature gradient as well as seasonally. Greater population densities of Aeromonas occurred below the oxygen chemocline when the lake was stratified. Survival of Aeromonas hydrophila under in situ conditions in both epilimnetic and hypolimnetic waters was determined through the use of polycarbonate membrane diffusion chambers during two separate reactor operating conditions. Survival levels of pure cultures of A. hydrophila corresponded to the distribution patterns of the naturally occurring Aeromonas-like populations. The greater survival of A. hydrophila during full reactor operation suggests that the fish populations may be exposed to Aeromonas for a longer period of time than when the reactor is not operating. PMID- 16345183 TI - Receptor site on clover and alfalfa roots for Rhizobium. AB - Sites on white clover and alfalfa roots that bind Rhizobium trifolii and R. meliloti capsular polysaccharides, respectively, were examined by fluorescence microscopy. Fluorescein isothiocyanate-labeled capsular material from R. trifolii bound specifically to root hairs of clover but not alfalfa. Binding was most intense at the root hair tips. Treatment of clover roots with 2-deoxyglucose (2 dG) prevented binding of R. trifolii capsular material to the roots. The sugar 2 dG enhanced the elution of clover root protein, which could bind to and specifically agglutinate R. trifolii but not R. meliloti or R. japonicum. The mild elution procedure left the roots intact. Agglutination of R. trifolii and passive hemagglutination of rabbit erythrocytes coated with the capsular material of R. trifolii were specifically inhibited by 2-dG. These results suggest that clover roots contain proteins that cross-link complementary polysaccharides on the surface of clover root hairs and infective R. trifolii through 2-dG-sensitive binding sites. Alfalfa root hairs were shown to specifically bind to a surface polysaccharide from R. meliloti. PMID- 16345184 TI - Semiquantitative Plate Assay for Determination of Cellulase Production by Trichoderma viride. AB - A plate clearing assay was devised to screen for high-producing cellulase mutants of Trichoderma viride. The method employs (i) the use of either rose bengal or oxgall to limit colony size and (ii) Phosfon D (tributyl-2, 4-dichloroben zylphosphonium chloride) to enhance cellulase detection, in combination with acid swollen cellulose on agar plates. The method was used to isolate constitutive cellulase mutants of T. viride and should prove useful for isolating high producing mutants from a range of organisms. This technique has been also used to determine the concentration at which glucose and glycerol inhibit cellulase synthesis by catabolite repression in the wild-type strains. PMID- 16345185 TI - Partial Purification and Some Properties of Tyrosine Phenol-Lyase from Aeromonas phenologenes ATCC 29063. AB - Tyrosine phenol-lyase was purified 32-fold from Aeromonas phenologenes ATCC 29063, the organism that produces phenol in refrigerated haddock. The purification procedure included ammonium sulfate fractionation, protamine sulfate treatment, and column chromatography with Sephadex G-200, diethyl-aminoethyl cellulose, and hydroxyapatite. The enzyme was found to be thermally inactivated at temperatures above 40 degrees C. The optimum pH of the enzyme was found to be pH 8.5. The Michaelis constants for l-tyrosine and pyridoxal phosphate were 2.3 x 10 M and 3.2 x 10 M, respectively. The molecular weight of tyrosine phenol-lyase was found by gel filtration and electrophoresis to be approximately 380,000. PMID- 16345186 TI - Production of gibberellic Acid using a dairy waste as the Basal medium. AB - Filtered whey has been shown to be a feasible basal medium for the production of gibberellic acid. A maximum yield of 750 mg/liter has been obtained, and the only supplement required was Mg ion at a final concentration of 10 mg/liter. PMID- 16345187 TI - Modification of membrane diffusion chambers for deep-water studies. AB - Membrane diffusion chambers as modified for deep-water lakes provided an improved handling ease, freedom from external damage, extended sampling life, good diffusivity, and excellent statistical reproducibility. PMID- 16345188 TI - Microbial transformations of selenium. AB - Resting cell suspensions of a strain of Corynebacterium isolated from soil formed dimethyl selenide from selenate, selenite, elemental selenium, selenomethionine, selenocystine, and methaneseleninate. Extracts of the bacterium catalyzed the production of dimethyl selenide from selenite, elemental selenium, and methaneseleninate, and methylation of the inorganic Se compounds was enhanced by S-adenosylmethionine. Neither trimethylselenonium nor methaneselenonate was metabolized by the Corynebacterium. Resting cell suspensions of a methionine utilizing pseudomonad converted selenomethionine to dimethyl diselenide. Six of 10 microorganisms able to grow on cystine used selenocystine as a sole source of carbon and formed elemental selenium, and one of the isolates, a pseudomonad, was found also to produce selenide. Soil enrichments converted trimethylselenonium to dimethyl selenide. Bacteria capable of utilizing trimethylselenonium, dimethyl selenide, and dimethyl diselenide as carbon sources were isolated from soil. PMID- 16345189 TI - Microbial decomposition of synthetic C-labeled lignins in nature: lignin biodegradation in a variety of natural materials. AB - Lignin biodegradation in a variety of natural materials was examined using specifically labeled synthetic C-lignins. Natural materials included soils, sediments, silage, steer bedding, and rumen contents. Both aerobic and anaerobic incubations were used. No C-labeled lignin biodegradation to labeled gaseous products under anaerobic conditions was observed. Aerobic C-labeled lignin mineralization varied with respect to type of natural material used, site, soil type and horizon, and temperature. The greatest observed degradation occurred in a soil from Yellowstone National Park and amounted to over 42% conversion of total radioactivity to CO(2) during 78 days of incubation. Amounts of C-labeled lignin mineralization in Wisconsin soils and sediments were significantly correlated with organic carbon, organic nitrogen, nitrate nitrogen, exchangeable calcium, and exchangeable potassium. PMID- 16345190 TI - Production and Purification of Thermostable Amylase and Protease of Thermomonospora viridis. AB - Maximum yields of amylase were produced by the thermophilic actinomycete Thermomonospora viridis in a modified Simpson and McCoy medium containing 1.5% corn starch and 0.5% mycological peptone with an initial pH 7.0. Best yields of amylase were obtained after incubation for 48 h, when the pH of the medium had risen to 8.2. Amylase was purified 313-fold by precipitation with n-propyl alcohol, dialysis against tap water, adsorption on Ca(3)(PO(4))(2), and fractionation on Sephadex G-100. Protease was produced in nutrient broth containing 0.5% starch and 1.0% corn steep liquor and at an initial pH 7.0. Maximum yields of protease were produced after 42 h. The protease was purified 54 fold by precipitation with n-propyl alcohol, dialysis against tap water, adsorption on Ca(3)(PO(4))(2), and fractionation on Sephadex G-200. PMID- 16345191 TI - Some Nutritional Characteristics of a Naturally Occurring Alga (Microcystis sp.) in a Guatemalan Lake. AB - The nutritional characteristics of an alga (Microcystis sp.) that occurs naturally in a Guatemalan lake were determined. The sun-dried material proved to have a high protein content (55.6%) and to be a possible good source of calcium and phosphorus (1, 169.1 and 633.4 mg/100 mg, respectively). Amino acid analysis showed that total sulfur amino acids were the most deficient ones, giving a protein score of 42 to the material. The in vitro protein digestibility of the material was 69.5%. Biological trials demonstrated that when the material was offered as the only protein source, very low consumption and a high mortality rate were obtained whether or not the diet was supplemented with 0.4% dl methionine. However, when the material supplied 25% of the total protein of a corn-algae diet, the protein quality of the cereal was significantly improved (P < 0.05). PMID- 16345192 TI - Methods for Growing Spirillum lipoferum and for Counting It in Pure Culture and in Association with Plants. AB - Methods are described for growing Spirillum lipoferum in quantities sufficient to serve as inoculant in field trials of its associative N(2)-fixing ability with higher plants and as a source of cells for the preparation of nitrogenase, cytochromes, respiratory enzymes, etc. A heavy inoculum of S. lipoferum grown on NH(4) was transferred to a medium of minimal nitrogen content, and initial rapid growth at the expense of residual combined nitrogen was replaced later by slower growth on N(2). Conversion to N(2) fixation was prompt upon exhaustion of fixed nitrogen; growth on N(2) was most rapid at a pO(2) of 0.005 to 0.007 atm. Numbers of S. lipoferum can be estimated by diluting soil, crushed roots, or other material, and inoculating diluted samples into a stagnant semisolid medium. Development of a characteristic subsurface layer of organisms and demonstration the these organisms can reduce C(2)H(2) are presumptive evidence that they are S. lipoferum. With most-probable-number tables the observations can be converted to numbers of S. lipoferum in the samples. The most-probable-number method indicated that numbers of S. lipoferum may increase 100-fold or more in roots of maize removed from the plant and incubated for 24 h at 30 degrees C at a pO(2) initially adjusted to 0.01 atm. PMID- 16345193 TI - Effects of Naturally Occurring Aquatic Organic Fractions on Am Uptake by Scenedesmus obliquus (Chlorophyceae) and Aeromonas hydrophila (Pseudomonadaceae). AB - Naturally occurring organics were extracted from water collected from Skinface Pond near Aiken, S.C. Organics were separated into four nominal diameter size fractions (I, >0.0183; II, 0.0183 to 0.0032; III, 0.0032 to 0.0009; IV, <0.0009 mum) by membrane ultrafiltration and introduced into Scenedesmus obliquus and Aeromonas hydrophila cultures to determine their effects on Am availability for uptake. Effects on Am uptake were determined in actively growing S. obliquus cultures after 96 h of growth and in dense cultures of nongrowing cells after 4 h. Uptake by A. hydrophila was determined after 4 and 24 h in actively growing cultures. All organic fractions stimulated S. obliquus growth, with the most pronounced effects due to larger organic fractions, whereas no apparent growth stimulation of A. hydrophila was observed for any organic fraction. For both long term and short-term studies, cellular Am concentration (picocuries/cell) increased with increasing Am concentration for S. obliquus and A. hydrophila. Fraction IV increased Am uptake by both S. obliquus and A. hydrophila during 4-h incubations. During 96-h incubations fraction I was flocculated and cosedimented, with S. obliquus and A. hydrophila cells causing an apparent increase in Am uptake. Fractions II and III reduced apparent Am uptake by S. obliquus as a result of biological dilution caused by increased algal growth due to the organics. Fraction IV caused a reduction in Am uptake by S. obliquus not attributable to biological dilution. Organics increased Am uptake by A. hydrophila during 4- and 24-h incubations. A. hydrophila also caused flocculation of fraction I during 96-h incubations. PMID- 16345194 TI - Effect of Temperature, Aeration, and Moisture on CO(2) Formation in Bench-Scale, Continuously Thermophilic Composting of Solid Waste. AB - A compost production system was employed to supply uniform material for controlled experiments of factorial design. Over a 96-h composting period, the cumulative amount of CO(2) evolved was maximal at 56 to 60 degrees C, an aeration rate that left an O(2) residual of 10 or 18% in the exhaust gas and a moisture content of 60% wet weight. Carbon dioxide evolution was submaximal at 64 degrees C and higher. PMID- 16345195 TI - Stability of Bacillus pumilus Spore Strips Used for Monitoring Radiation Sterilization. AB - [This corrects the article on p. 999 in vol. 31.]. PMID- 16345196 TI - Stability of lactic Acid bacteria to freezing as related to their Fatty Acid composition. AB - The viability of Streptococcus lactis and Lactobacillus sp. A-12 after freezing at -17 degrees C for 48 h was better preserved when the cells were grown in medium supplemented with oleic acid or Tween 80 (polyoxyethylene sorbitan monooleate). A pronounced change in the cellular fatty acid composition was noted when the bacteria were grown in the presence of Tween 80. In S. lactis the ratio of unsaturated to saturated fatty acids increased from 1.18 to 2.55 and in Lactobacillus sp. A-12 it increased from 0.85 to 1.67 when Tween 80 was added to the growth medium. The antibiotic cerulenin markedly inhibited the growth of lactic acid bacteria in tomato juice (TJ) medium but had almost no effect on the growth of the bacteria in TJ medium containing Tween 80 (or oleic acid). The antibiotic inhibited markedly the incorporation of [1-C]acetate but had no inhibitory effect on the incorporation of exogenous [1-C]oleate (or [1 C]palmitate) into the lipid fractions of lactic acid bacteria. Thus, the fatty acid composition of lactic acid bacteria, inhibited by the antibiotic cerulenin, can be modulated by exogenously added oleic acid (or Tween 80) without the concurrent endogenous fatty acid synthesis from acetate. The data obtained suggest that cerulenin inhibits neither cyclopropane fatty acid synthesis nor elongation of fatty acid acyl intermediates. The radioactivity of cells grown in the presence of [1-C]oleate and cerulenin was associated mainly with cyclopropane Delta19:0, 20:0 + 20:1, and 21:0 acids. As a consequence, cerulenin caused a decrease in the ratio of unsaturated to saturated fatty acids in lactic acid bacteria as compared with cells grown in TJ medium plus Tween 80 but without cerulenin. Cerulenin caused a decrease in the viability of S. lactis and Lactobacillus sp. A-12 after freezing at -17 degrees C for 48 h only when Tween 80 was present in the growth medium. We conclude that the sensitivity of lactic acid bacteria to damage from freezing can be correlated with specific alterations in the cellular fatty acids. PMID- 16345197 TI - Factors affecting the cellulolytic activity of rumen contents. AB - The cellulolytic activity of rumen contents was assayed by measuring losses in weight and tensile strength of cotton yarn incubated in rumen contents in the presence of dietary additives (barley, tallow) and at different pH values. The addition of barley depressed cellulolysis and the titer of filter paper-degrading bacteria only if the pH was allowed to fall. Lowering the pH from 7.0 to 6.0 by addition of HCl almost completely inhibited attack of cotton and greatly reduced the titer of filter paper-degrading bacteria. The layering of tallow on cotton inhibited attack of cotton but did not decrease the titer of filter paper degrading bacteria. The results are discussed with special reference to the importance of the study of cellulosic substrates, which require a known enzyme or mixture of enzymes for attack. PMID- 16345198 TI - Properties of an immobilized pesticide-hydrolyzing enzyme. AB - A bacterial enzyme(s) capable of hydrolyzing nine organophosphate insecticides was covalently bound to glass. The efficiency of this binding reaction ranged from 4 to 17%. Under continuous column operation, the immobilized enzyme(s) had an extrapolated half-life of 280 days. The specific activity of this glass covalently bound hydrolase activity for parathion varied from 0.035 to 0.15 mumol/min per g of glass. The bound activity increased with decreasing glass particle size; however, the flow resistance also increased. Immobilized enzyme(s) kinetics were approximately 50% slower than those of the free enzyme(s), but there was no significant difference in the effect pH and temperature had on the activity of immobilized and free enzyme(s). PMID- 16345199 TI - Purification and Some Biological Properties of Asparaginase from Azotobacter vinelandii. AB - Asparaginase was found in the soluble fraction of cells of Azotobacter vinelandii, and its activity remained the same during growth of the organism in a nitrogen-free medium. The specific activity and the yield of A. vinelandii increased twofold in the presence of ammonium sulfate. Within limits, the temperature (30 to 37 degrees C) and pH (6.5 to 8.0) of the medium showed little effect on the levels of enzyme activity. The enzyme was purified to near homogeneity by standard methods of enzyme purification, including affinity chromatography, and had optimum activity at pH 8.6 and 48 degrees C. The approximate molecular weight was 84,000. The apparent K(m) value for the substrate was 1.1 x 10 M. Metal ions or sulfhydryl reagents were not required for enzyme activity. Cu, Zn, and Hg showed concentration-dependent inhibition, whereas amino and keto acids had no effect on the enzyme activity. Asparaginase was stable when incubated with rat serum and ascites fluid. The enzyme had no effect on the membrane of sheep erythrocytes and did not inhibit the incorporation of radioactive precursors into deoxyribonucleic acid, ribonucleic acid, and protein in Yoshida ascites sarcoma cells. Asparaginase activity was not detected in the tumor cells. PMID- 16345200 TI - Form of Vitamin B(12) and Its Role in a Methanol-Utilizing Bacterium Protaminobacter ruber. AB - A methanol-utilizing bacterium, Protaminobacter ruber, produced a large amount of vitamin B(12). The compounds were isolated from the cells and identified as methylcobalamin (methyl-B(12)) and adenosylcobalamin (adenosyl-B(12)) by various tests. The variation in the form of B(12) during cultivation was examined by bioautography with cellulose acetate membrane electrophoresis. Methyl-B(12) and adenosyl-B(12) were the two main B(12) compounds produced in the various phases of bacterial growth. The ratio of the amount of methyl-B(12) to total B(12) compounds was higher during the earlier phases of growth. After the logarithmic phase, adenosyl-B(12) was the predominant form. The existence of N methyltetrahydrofolate:homocysteine transmethylase and methyl-B(12):homocysteine transmethylase was demonstrated in cell-free extracts of Protaminobacter ruber. Methyl-B(12) in P. ruber seems to function mainly in the B(12)-dependent methionine synthetase system. PMID- 16345201 TI - Production of Kestoses (Fructosylsucroses) by Phytophthora parasitica var. nicotianae. AB - The synthesis of kestoses (trisaccharides composed of two fructose units and one glucose unit) by races 0 and 1 of Phytophthora parasitica var. nicotianae is shown. The trisaccharide is found in culture filtrates of isolates grown in liquid media containing 3% sucrose. The utilization of sucrose and trisaccharide formation by the organisms over a 16-day period is described. The kestoses were identified by chemical and enzymatic analysis, and two of three possible isomers were found. PMID- 16345202 TI - Transparent plastic incubator for the anaerobic glove box. AB - An incubator designed for use inside an anaerobic glove box is described. The incubator is made of transparent plastic material, has sliding plastic doors, and can be made in various sizes from readily available materials. PMID- 16345203 TI - Use of adenosine 5'-triphosphate as an indicator of the microbiota biomass in rumen contents. AB - A number of techniques were tested for their efficiency in extracting adenosine 5'-triphosphate (ATP) from strained rumen fluid (SRF). Extraction with 0.6 N H(2)SO(4), using a modification of the procedure described by Lee et al. (1971), was the most efficient and was better suited for extracting particulate samples. Neutralized extracts could not be stored frozen before assaying for ATP because large losses were incurred. The inclusion of internal standards was necessary to correct for incomplete recovery of ATP. The ATP concentration in rumen contents from a cow receiving a ration of dried roughage (mainly alfalfa hay) ranged from 31 to 56 mug of ATP per g of contents. Approximately 75% of the ATP was associated with the particulate material. The ATP was primarily of microbial origin, since only traces of ATP were present in the feed and none was found in "cell-free" rumen fluid. Fractionation of the bacterial and protozoal populations in SRF resulted in the isolation of an enriched protozoal fraction with a 10-fold higher ATP concentration than that of the separated rumen bacteria. The ATP pool sizes of nine functionally important rumen bacteria during the exponential phase of growth ranged from 1.1 to 17.6 mug of ATP per mg of dry weight. This information indicates that using ATP as a measure of microbial biomass in rumen contents must be done with caution because of possible variations in the efficiency of extraction of ATP from rumen contents and differences in the concentration of ATP in rumen microbes. PMID- 16345204 TI - Bacteriological investigation of alberta meat-packing plant wastes with emphasis on salmonella isolation. AB - The waste treatment facilities and final effluents of 11 meat-packing plants in the Province of Alberta were investigated primarily to determine the numbers of indicator bacteria and the presence of Salmonella. This was done to discover the efficiency of the treatment systems presently in operation in reducing bacterial numbers and to establish the need for disinfection and for bacterial standards for these effluents. Data obtained showed that the final effluents were of very poor quality bacteriologically, with numbers of indicator organisms comparable to those found in raw sewage. Primary treatment facilities were ineffective in reducing the numbers of these bacteria. The secondary treatment facility investigated achieved greater than a 99% reduction of indicator bacteria. Salmonella were isolated from the final effluents of 78% of the plants, including the plant using secondary treatment. In total, 21 Salmonella serotypes were isolated. Salmonella isolates were not antibiotic resistant, but certain coliform and fecal coliform isolates demonstrated resistance to chloramphenicol, tetracycline, and ampicillin. PMID- 16345205 TI - Control of Sexual Reproduction in Gibberella zeae (Fusarium roseum "Graminearum"). AB - Sexual reproduction in Gibberella zeae (Fusarium roseum) is regulated by the fungal sex hormone zearalenone, which is known to be synthesized only by species of Fusarium. The presence of cyclic adenosine 3',5'-monophosphate (cAMP) in mycelium of this fungus has been confirmed by analyses with thin-layer and gas liquid chromatography, fluorescent properties, ultraviolet absorption, competitive protein-binding tests, and degradation by cyclic phosphodies-terase. cAMP but not cyclic guanosine monophosphate increased both the number of perithecia formed and the incorporation of [1-C]acetate into zearalenone. It is proposed that cAMP stimulates the synthesis of zearalenone which then exerts its effect directly or indirectly on formation of perithecia. PMID- 16345206 TI - Limax amoebae in public swimming pools of albany, schenectady, and rensselaer counties, new york: their concentration, correlations, and significance. AB - A survey was conducted on 30 halogenated public swimming pools, located in Albany, Schenectady, and Rensselaer counties, to determine their open-water limax amoeba densities. Six were outdoor pools. Other variables measured were the standard plate count, total seston, free residual chlorine or bromine, total alkalinity, total hardness, orthophosphate, total soluble phosphorus, specific conductance, pH, temperature, and several engineering parameters including the rate and type of filtration as well as a saturation index. Amoebae were isolated on agar plates at 37 degrees C using heat-killed bacterial suspensions of Enterobacter cloacae or Escherichia coli. Most probable number estimates of amoebic densities ranged from not detectable (<0.01) to 110 amoebae per liter. The median concentration of amoebae was 0.9/liter. Eighty percent of the pools examined had less than 5 amoebae per liter. Significant correlations (P < 0.05) were found between amoebic densities and the log(10) of the standard plate count, orthophosphate, and total soluble phosphorus. No significant difference was found between amoebic densities in outdoor and indoor pools. Preliminary tests for the presence of the human pathogen Naegleria fowleri were inconclusive. PMID- 16345207 TI - Relationship of vaginal cytology to alterations of the vaginal microflora of rats during the estrous cycle. AB - The influence of cyclic changes occurring in the vaginal tract during the estrous cycle upon the indigenous microflora of the vagina has been investigated by semiquantitative techniques in virgin female rats. By plate counts performed on material lavaged daily from the vaginal tract of several rats, it is apparent that bacterial counts are elevated in the proestrus and estrus phases of the cycle to values several orders of magnitude greater than those observed during metestrus or diestrus phases. Increases in vaginal bacterial counts were associated with the presence of cornified epithelial cells in the vagina; these cells were predominantly nonviable. Decreases in the vaginal bacterial content were related to the influx of leukocytes into the vagina after estrus. When leukocytes were present in the vaginal tract, they were 90 to 100% viable. From these observations it has been concluded that the female vaginal tract and the bacteria which colonize it represent a dynamic ecosystem which is responsive to cyclic events occurring in the estrous cycle. The changing cellular content of the vaginal tract may have relevance to the observed cyclic changes in the bacterial content of the vagina. PMID- 16345208 TI - Occurrence of Escherichia coli in Wild Cottontail Rabbits. AB - Free-ranging cottontail rabbits (Sylvilagus floridanus) from two areas in central Pennsylvania were sampled over a 4-year period. Large numbers of coliforms were isolated from the intestinal tracts of these animals; in 136 of the 141 rabbits sampled, Escherichia coli was found to be a major component of the alimentary flora. Four serogroups (O7, O77, O73, and O103) were predominant among the isolates and were considered resistant coliflora of this species of cottontail rabbit. PMID- 16345209 TI - Mutant of Methylomonas methanolica and Its Characterization with Respect to Biomass Production from Methanol. AB - A stable variant of Methylomonas methanolica, with a higher temperature optimum for growth, was obtained after mutagenic treatment and selection. The mutant strain M13V has an optimum growth temperature of 35 to 37 degrees C and a maximum at 43 degrees C, as compared with 30 and 40 degrees C for the wild strain. Strain M13V and M. methanolica have similar basic characteristics and cell composition. An extracellular polysaccharide is produced by both strains, but this property is more pronounced in strain M13V. In strain M13V the production is favored by high temperature, low growth rate, and oxygen limitation. In continuous culture of strain M13V, the polysaccharide production was partly growth associated and partly independent of the growth rate. The extracellular polysaccharide acted as a flocculating agent. A relationship between polysaccharide concentration and sedimentation rate was found. Biomass production from strain M13V is most effective at 35 degrees C with respect to both growth rate and substrate utilization. It was found that the yield coefficient for methanol was independent of the dilution rate, whereas the yield coefficient for oxygen increased and the production coefficient for carbon dioxide decreased at increasing dilution rates. These results are discussed in connection with the polysaccharide production. PMID- 16345210 TI - Lipid accumulation in an oleaginous yeast (Candida 107) growing on glucose under various conditions in a one- and two-stage continuous culture. AB - Lipid accumulation and fatty acid composition in Candida 107 have been studied using a two-stage continuous culture system in which the first vessel was run under carbon-limited conditions and then the entire output was passed into a second vessel, where lipid accumulation was stimulated by adding only glucose. Maximum lipid accumulation (28% of yeast [dry weight]) occurred for a volume ratio of vessel 1 to vessel 2 of 3:5, with 30 g of glucose per liter being added to vessel 2 operated at 25 degrees C with an aeration rate of between 0.1 and 1.0 volume of air/volume of medium per min. Although the maximum specific rate of lipid formation (0.05 g of lipid/g of yeast per h) was higher than in a nitrogen limited, single-stage system, the efficiency of lipid formation was much less and never exceeded 14 g of lipid produced per 100 g of glucose consumed. The fatty acid composition was not significantly altered in either the two-stage or single stage culture (nitrogen-limited) systems by changes in growth temperature (from 19 to 33 degrees C) or aeration rates (0.05 to 1.0 volume of air/volume of medium per min); or, in the two-stage system, by changes in the residence time of the yeast in the second vessel (from 3.2 to 24.4 h), or, in the single-stage system, by changes in pH (from 3.5 to 7.5). Only when the concentration of glucose entering vessel 2 of the two-stage system was less than 30 g/liter did significant changes in the fatty acids occur. Thus, although a two-stage continuous culture system allows lipid accumulation to be separated from the growth phase, it offers no practical advantages over a single-stage system as a means of producing microbial oils and fats. PMID- 16345211 TI - Effects of varying concentrations of novobiocin incorporated into two salmonella plating media on the recovery of four enterobacteriaceae. AB - Hydrogen sulfide-producing strains of salmonellae, Escherichia coli, Citrobacter freundii, and Proteus mirabilis were isolated from fresh pork sausage. All the strains produced black-centered colonies on Hektoen enteric agar (HE). On xylose lysine deoxycholate agar (XLD), C. freundii produced yellow colonies, and the strains of the other three genera formed black-centered colonies. The selectivity of HE and XLD for salmonellae was improved by the addition of novobiocin to both media. With increasing concentrations of novobiocin, the degree of growth inhibition for the four genera was less on HE than on XLD. Novobiocin concentrations of 80 mug/ml in HE and 5 mug/ml in XLD did not affect the growth or colonial morphology of salmonellae. When 80 mug of novobiocin per ml was incorporated into HE, P. mirabilis strains were not recovered, 40% of C. freundii strains failed to form black-centered colonies, and growth of E. coli strains was not affected but colonies were altered without eliminating the black centers. When novobiocin at 5 mug/ml was incorporated into XLD, colonies of P. mirabilis strains were not recovered, C. freundii formed yellow colonies, and the colonies of the H(2)S-producing E. coli strains were unaffected. PMID- 16345212 TI - Evaluation of recovery methods to detect coliforms in water. AB - Various recovery methods used to detect coliforms in water were evaluated by applying the membrane filter chamber technique. The membrane filter chambers, containing pure-culture suspensions of Escherichia coli or natural suspensions of raw sewage, were immersed in the stream environment. Samples were withdrawn from the chamber at regular time intervals and enumerated by several detection methods. In general, multiple-tube fermentation techniques gave better recovery than plating or membrane filtration procedures. The least efficient method of recovery resulted when using membrane filtration procedures, especially as the exposure period of the organisms to the stream environment increased. A 2-h enrichment on a rich, nonselective medium before exposure to selective media improved the recovery of fecal coliforms with membrane filtration techniques. Substantially enhanced recoveries of E. coli from pure-culture suspensions and of fecal coliforms from raw-sewage suspensions were observed when compared with recoveries obtained by direct primary exposure to selective media. Such an enrichment period appears to provide a nontoxic environment for the gradual adjustment and repair of injured cells. PMID- 16345213 TI - Comparison of Nitrogen Fixation Activity in Tall and Short Spartina alterniflora Salt Marsh Soils. AB - A comparison of the N(2) fixers in the tall Spartina alterniflora and short S. alterniflora marsh soils was investigated. Zero-order kinetics and first-order kinetics of acetylene reduction were used to describe the activity of the N(2) fixers in marsh soil slurries. It was found that the V(max) values were approximately 10 times greater for the N(2) fixers in the tall Spartina than in the short Spartina marsh when raffinose was used as the energy source. In addition, the (K(s) + S(n)) values were approximately 4 to 15 times lower for the N(2) fixers in the tall Spartina than in short Spartina marsh. First-order kinetics of nitrogen fixation for several substrates indicate that the N(2) fixers in the tall Spartina marsh were two to seven times more active than those in the short Spartina marsh. Ammonium chloride (25 mug/ml) did not inhibit nitrogen fixation in the tall Spartina marsh, but there was a 50% inhibition in nitrogen fixation in the short Spartina marsh. On the other hand, sodium nitrate inhibited nitrogen fixation almost 100% at 25 mug/ml in both soil environments. Amino nitrogen (25 to 100 mug/ml) had little or no effect on nitrogen fixation. The results indicate that the N(2) fixers in the tall Spartina marsh were physiologically more responsive to nutrient addition than those in the short Spartina marsh. This difference in the two populations may be related to the difference in daily tidal influence in the respective areas and thus provide another explanation for the enhanced S. alterniflora production in the creek bank soil system. PMID- 16345214 TI - The environment and the microbial ecology of human skin. AB - Microbial flora of the skin of three human population groups representing different natural environments was examined quantitatively and qualitatively to determine whether environmental differences in temperature and humidity can influence the microbial flora of normal skin. Five anatomical skin sites - hands, back, axillae, groin, and feet - were sampled from 10 subjects working in a high humidity, high-temperature environment, 10 subjects from a low-temperature, high humidity environment, and 10 subjects working in a moderate-temperature and low humidity environment. Bacterial populations were significantly larger from the back, axillae, and feet in individuals from the high-temperature and high humidity environment as compared to the moderate-temperature, low-humidity environment. High humidity and low temperature had no significant effect on total populations, but this group showed a higher frequency of isolation of fungi, and gram-negative bacteria from the back and feet. Although there was an indication that increase in the environmental humidity could result in an increased frequency of isolation of gram-negative bacteria, there was no evidence that an increase in either temperature or humidity altered the relative proportions of gram-negative bacteria in the predominantly gram-positive microbial flora found on normal skin. It was concluded that, although climatic changes may cause fluctation in microbial populations from certain sites, they are not a major influence on the ecology of the microbial flora of normal skin in the natural environment. The variables introduced by studying individuals in their natural environment and the influence of these on the results are discussed. PMID- 16345215 TI - Virus and bacteria removal from wastewater by rapid infiltration through soil. AB - A rapid infiltration land wastewater application site, composed of unconsolidated silty sand and gravel, which has been in continuous operation for over 30 years was examined for the accumulation and/or migration of a tracer virus (coliphage f2), indigenous enteroviruses, and enteric indicator bacteria in the soils and underlying groundwater. Tracer f2 penetrated into groundwater together with the front of percolating primary effluent and was not observed to concentrate on the upper soil layers. The tracer virus concentration in a 60-foot (about 18.3-m) deep observation well directly beneath the wastewater application area began to increase within 48 h after application to the soil. The tracer level in this well stabilized after 72 h at a level of approximately 47% of the average applied concentration. Indigenous enteroviruses and tracer f2 were sporadically detected in the groundwater at horizontal distances of 600 feet (about 183 m) from the application zone. Laboratory soil adsorption studies confirmed the poor virus adsorption observed at the site. This was especially true on surface soils when contained in wastewater. Enteric indicator bacteria were readily concentrated on the soil surface by filtration on the soil surface mat. However, during tracer f2 virus tests, comparison studies with fecal Streptococcus revealed that bacteria capable of penetrating the surface were able to migrate into the groundwater. They were detected at the same locations as tracer and enteric viruses. PMID- 16345216 TI - Modified radioimmunoassay determination for staphylococcal enterotoxin B in foods. AB - The sensitivity of solid-phase radioimmunoassay for the measurement of staphylococcal enterotoxin B (SEB) in foods was decreased by food constituents that react with rabbit anti-SEB with an equivalency of over 2 ng/ml. This activity was minimized by a conditioning step for anti-SEB and by removal of interfering compounds in the sample by extraction. The assay was a sequential solid-phase radioimmunoassay technique in which polystyrene test tubes were initially incubated with antisera and then with bovine serum albumin. The tubes were then conditioned with either a centrifuged aqueous cheese extract or, equally effective, reconstituted nonfat dry milk for 16 h at 4 degrees C. Samples of milk or heat-treated and CHCl(3)-extracted cheese or chicken salad slurries were incubated in the assay tubes for 6 h at 37 degrees C. The samples were replaced by I-labeled SEB and incubated for a further 2 to 4 h before the contents were removed and the tubes were washed and counted. A buffer solution containing known concentrations of toxin served as standards for assaying SEB in the food extracts. The entire assay can be accomplished within 24 h with a sensitivity of 1 ng/ml in milk and in the cheese extract or 1.3 ng/ml in the chicken salad extract. PMID- 16345217 TI - Acetylene reduction by soil cores of maize and sorghum in Brazil. AB - Nitrogenase activity was measured by the C(2)H(2) reduction method in large soil cores (29 cm in diameter by 20 cm in depth) of maize (Zea mays) and sorghum (Sorghum vulgare). The activity was compared to that obtained by a method in which the roots were removed from the soil and assayed for nitrogenase activity after an overnight preincubation in 1% O(2). In a total of six experiments and 28 soil cores, the nitrogenase activity of the cores was an average of 14 times less than the activity of roots removed from the same cores and preincubated. Nitrogenase activity in the cores was very low and extrapolated to an average nitrogen fixation rate of 2.8 g of N/hectare per day. It was shown that inadequate gas exchange was not a reason for the lower activity in the soil cores, and the core method gave satisfactory results for nitrogenase activity of soybeans (Glycine max) and Paspalum notatum. PMID- 16345218 TI - Timed-release capsule method for the detection of salmonellae in foods and feeds. AB - A new method was developed for the detection of injured and uninjured salmonellae in foods and feeds. The steps of pre-enrichment in a nonselective broth and selective enrichment in a selective medium were combined into a single procedure. This was achieved by the gradual release of selective agents from wax-coated gelatin capsules added at the time of inoculation of nonselective basal broths. Pre-enrichment in lactose broth was combined with selective enrichment in tetrathionate or selenite-cystine broth by using timed-release capsules containing iodine or selenite. Five different categories of foods and feeds, naturally contaminated with salmonellae, were examined to compare the efficiencies of the capsule methods with conventional procedures. Combination of the separate steps of pre-enrichment and selective enrichment into a single procedure was feasible and resulted in substantial savings of labor and materials. PMID- 16345219 TI - Survival of a psychrophilic marine Vibrio under long-term nutrient starvation. AB - Ant-300, a psychrophilic marine vibrio isolated from the surface water of the Antarctic convergence, was starved for periods of more than 1 year. During the first week of starvation, cell numbers increased from 100 to 800% of the initial number of cells. Fifty percent of the starved cells remained viable for 6 to 7 weeks while a portion of the population remained viable for more than 1 year. During the first 2 days of starvation, the endogenous respiration of the cells decreased over 80%. After 7 days, respiration had been reduced to 0.0071% total carbon respired per hour and remained constant thereafter. After 6 weeks of starvation, 46% of the cellular deoxyribonucleic acid had been degraded. Observation of the cellular deoxyribonucleic acid with Feulgen staining before starvation showed the average number of nuclear bodies per cell varied from 1.44 to 4.02 depending on the age of the culture. A linear relationship was found between the number of nuclear bodies per cell and the increase in cell numbers upon starvation. Our data suggest that Ant-300 is capable of surviving long periods of time with little or no nutrients and is therefore well adapted for the sparse nutrient conditions of the colder portions of the open ocean. PMID- 16345220 TI - Retrieval of concentrated and undecompressed microbial populations from the deep sea. AB - A device for sampling at depths of up to 6,000 m is described in which 3 liters of seawater is concentrated over a Nucleopore filter to about 13 ml and retrieved under in situ pressure and temperature. Subsamples can be withdrawn into transfer units that are equipped with individual gas accumulators for preventing loss of pressure during prolonged periods of storage. Transfer of samples or sample portions into sterile medium contained in pre-pressurized incubation vessels and continued subsampling therefrom permit time course experiments for the study of natural populations of deep-sea microorganisms in the absence of decompression. A test experiment with a water sample from a depth of 2,600 m supplemented with radioactively labeled Casamino Acids showed reduced rates of substrate incorporation and respiration as compared with data from a decompressed control. The barotolerance observed in this study was characterized by reduced, rather than equal, activities recorded at elevated pressures as compared with 1-atm controls. PMID- 16345221 TI - Continuous open flow-through system as a model for oil degradation in the arctic ocean. AB - A continuous flow-through system incubated in situ was used to model oil biodegradation in Arctic coastal waters. High numbers of oil-degrading microorganisms were found in the Arctic coastal waters examined in this study. The microbial community underlying oil slicks increased and showed a population shift to a greater percentage of hydrocarbon-utilizing microorganisms. Microbial populations and oil biodegradation were increased by the addition of nitrogen and phosphorus. Both abiotic and biodegradative losses were lower than expected, perhaps due to the unusually harsh, ice-dominated Arctic summer, during which these tests were conducted. Chromatographic and spectrometric analyses showed that residual oils contained similar percentages of individual components and classes of hydrocarbons, regardless of the amount of degradation, indicating that most components of the oil were being degraded at similar rates. PMID- 16345222 TI - Dietary effects on the composition of fecal flora of rats. AB - A long-term animal feeding experiment was conducted to compare the effect of meat and Wayne laboratory chow diets on the composition of rat fecal flora. Fecal bacteria were enumerated on selective media under both aerobic and anaerobic conditions. Intrarectal administration of N-methyl-N' -nitro-N-nitro-soguanidine (MNNG) affected the count only on the phenylethylalcohol and veillonella-neomycin agars, whereas a slightly higher number of anaerobes appeared in the feces of rats that were treated with MNNG as compared with those obtained in the feces of untreated rats on the meat diet. In the absence of MNNG, feces of meat-fed rats yielded higher bacterial counts on aerobically incubated MacConkey agar, deoxycholate agar, and Pfizer selective enterococcus agar as well as higher numbers of clostridia on anaerobic egg yolk agar than did feces of rats on the Wayne diet. Feces of the group fed the Wayne diet produced more colonies on aerobic mitis-salivarius agar and lactobacillus agar as well as on anaerobically incubated phenylethylalcohol agar, veillonellaneomycin agar, bifidobacteria agar, fusobacterium (Nissui) agar, and kanamycin-vancomycin blood agar. These differences were consistent throughout the 1-year feeding period. PMID- 16345223 TI - Bacterial growth kinetics on diphenylmethane and naphthalene-heptamethylnonane mixtures. AB - Experiments were carried out to determine if diphenylmethane is utilized by a species of Pseudomonas (Hydrogenomonas) in the dissolved state regardless of the physical state (liquid or solid) of the undissolved diphenylmethane suspended in the medium. Bacterial growth rates in the presence of various amounts of solid or liquid diphenylmethane indicate that liquid diphenylmethane is utilized at the aqueous-diphenylmethane interface but that solid diphenylmethane is not. A Pseudomonas sp. that was isolated on naphthalene (solid), but could not utilize heptamethylnonane, was grown in the presence of various amounts of a naphthalene heptamethylnonane mixture (liquid). The growth rates indicate that the bacterium could utilize naphthalene at the aqueous-hydrocarbon interface, which is not the case in the absence of the heptamethylnonane. PMID- 16345224 TI - Multisurface glass roller bottle for growth of animal cells in culture. AB - A multisurface glass roller bottle has been constructed for the growth of animal cells in culture. This bottle contains five concentrically placed glass cylinders that provide additional surfaces for the growth of animal cells. The bottle occupies the same space as a standard roller bottle, but it contains nine times the surface area of a standard bottle. L and HeLa cells can be grown in the bottle with cell yields 5- to 10-fold greater than in a standard bottle. L cells can be induced to produce interferon in the multisurface bottle. PMID- 16345225 TI - Resistance of cysts of amoebae to microbial decomposition. AB - Cyst walls of Acanthamoeba rhysodes, A. palestinensis, A. castellanii, and one other strain of Acanthamoeba contain 36 to 45% protein and 20 to 34% carbohydrate. More than half of the protein in the walls of A. palestinensis, A. castellanii and Acanthamoeba sp. is accessible to and hydrolyzed by protease, and 67 to 69% of the carbohydrate of A. palestinensis and A. rhysodes walls is hydrolyzed by cellulase. The extent of hydrolysis of walls of the other amoebae by these enzymes is appreciably less, and chitinase and beta-1,3-glucanase have no detectable effect. Protease solubilizes 10% or less of the weight of intact cysts, and no solubilization is observed with cellulase. Walls of A. palestinensis are extensively degraded in soil, the activity is less with A. rhysodes, and little attack on the other amoebae occurs. When added to soil, the protozoa excyst and grow for short periods, the trophozoites then die, and chiefly cysts persist thereafter. PMID- 16345226 TI - Microtechnique for most-probable-number analysis. AB - A microtechnique based on the most-probable-number (MPN) method has been developed for the enumeration of the ammonium-oxidizing population in soil samples. An MPN table for a research design ([8 by 12] i.e., 12 dilutions, 8 replicates per dilution) is presented. A correlation of 0.68 was found between MPNs determined by the microtechnique and the standard tube technique. Higher MPNs were obtained with the microtechnique with increased accuracy in endpoint determinations being a possible cause. Considerable savings of time, space, equipment, and reagents are observed using this method. The microtechnique described may be adapted to other microbial populations using various types of media and endpoint determinations. PMID- 16345227 TI - Sensitivity of Various Bacteria, Including Actinomycetes, and Fungi to Cadmium and the Influence of pH on Sensitivity. AB - A variety of microorganisms, including gram-negative and gram-positive eubacteria, actinomycetes, yeasts, and filamentous fungi, were tested for their sensitivity to cadmium (Cd). In general, the actinomycetes were more tolerant to Cd than were the eubacteria; gram-negative eubacteria were more tolerant to Cd than were gram-positive eubacteria. The period of exponential growth of the eubacteria and actinomycetes was extended in the presence of Cd. Wide extremes in sensitivity to Cd were noted among the fungi; there was no correlation between the class of fungus and tolerance to Cd. Fungal sporulation was more sensitive to Cd than was mycelial growth, as spore formation was inhibited at Cd concentrations that were noninhibitory to mycelial proliferation. The toxicity of Cd to the eubacteria, actinomycetes, and fungi appeared to be pH dependent, as toxicity was generally potentiated at pH 8 or 9. PMID- 16345228 TI - Reductions in the toxicity of cadmium to microorganisms by clay minerals. AB - The clay minerals montmorillonite and kaolinite protected bacteria, including actinomycetes, and filamentous fungi from the inhibitory effects of cadmium (Cd). Montmorillonite provided greater protection than did equivalent concentrations of kaolinite. The protective ability of the clays was correlated with their cation exchange capacity (CEC). The greater the CEC, the greater the absorbancy of exogenous Cd by the exchange complex and the greater the protection. The greater protection afforded by montmorillonite, as compared to kaolinite, was correlated with its higher CEC. Clays homoionic to Cd did not protect against exogenous Cd, as the exchange complex was already saturated with Cd. Montmorillonite homoionic to Cd was more detrimental to microbial growth than was kaolinite homoionic to Cd, as more Cd was present on and apparently desorbed from the montmorillonite. PMID- 16345229 TI - Application of Oxygen-Enriched Aeration in the Conversion of Glycerol to Dihydroxyacetone by Gluconobacter melanogenus IFO 3293. AB - Gluconobacter melanogenus 3293 converts glycerol to dihydroxyacetone(DHA) during exponential growth on a yeast extract-phosphate medium at pH 7. The efficiency of this conversion in 25-liter batch fermentations has been found to increase over threefold, when oxygen tension is controlled by increasing the partial pressure of oxygen in the aeration. Conversion of glycerol to DHA does not occur under oxygen-limited fermentation conditions. When the dissolved oxygen tension was maintained at 0.05 atmospheres (using oxygen-enriched air), quantitative conversion of up to 100 g of glycerol/liter to DHA was obtained in 33 h. The amount of glycerol converted can be increased without increasing impeller speed or aeration rate. This increase is not the result of increased production of cell mass. The specific conversion of glycerol to DHA increased from 12.2 g of DHA/g of cell mass at the point of maximum conversion to 35.8 with oxygen enrichment. This increased specific production occurred even though the specific growth rate during the period of oxygen enrichment decreased from 0.23 to 0.06/h. PMID- 16345230 TI - Tentative identification of methanogenic bacteria by fluorescence microscopy. AB - Methanogenic bacteria, which are presently identified on the basis of cell morphology and substrate conversion to CH(4), can be differentiated from nonmethanogens and identified in pure or mixed culture on the basis of their autofluorescence under ultraviolet illumination. PMID- 16345231 TI - Characterization of a Cytoplasmic Reserve Glucan from Ruminococcus albus. AB - Ruminococcus albus, an anaerobic bacterium that digests cellulose in the rumen of cattle, produces intracellular polysaccharide granules varying from 0.05 to 0.31 mum in diameter when grown in batch culture. This polysaccharide material was purified and found to contain d-glucose as the only reducing sugar. The polyglucose polymer was slightly opalescent in aqueous solution and formed a strong reddish purple iodine complex with a maximum absorbance at 550 nm. Its infrared spectrum had characteristic absorption bands at 8.70, 9.25, and 9.75 mum and was identical with that of the amylopectin-glycogen type of Megasphaera elsdenii and that of the glycogen of enteric bacteria and beef liver. It reacted strongly with concanavalin A. Methylation analysis showed that the glucan contained 2,3,4,6-tetra-O-MeG-2,3,6-Tri-O-MeG-2,3-Di-O-MeG in a ratio of 8:84:8. Characterization of the products obtained by treatment with isoamylase indicates that the glucan of R. albus is of the glycogen type. PMID- 16345232 TI - Extradiol cleavage of 3-methylcatechol by catechol 1,2-dioxygenase from various microorganisms. AB - The isofunctional enzymes of catechol 1,2-dioxygenase from species of Acinetobacter, Pseudomonas, Nocardia, Alcaligenes, and Corynebacterium oxidize 3 methylcatechol according to both the intradiol and extradiol cleavage patterns. However, the enzyme preparations from Brevibacterium and Arthrobacter have only the intradiol cleavage activity. Comparison of substrate specificity among these isofunctional dioxygenases shows striking differences in the oxidation of 3 methylcatechol, 4-methylcatechol and pyrogallol. PMID- 16345233 TI - Induction and Catabolite Repression of Glucosidases in Pseudomonas maltophilia. AB - Intracellular alpha-and beta-glucosidases were induced in cell suspensions of Pseu-domonas maltophilia by maltose or cellobiose, and the synthesis of these enzymes was sensitive to apparent catabolite repression by alpha-ketoglutarate. PMID- 16345234 TI - Production of the Tremorgenic Mycotoxins Verruculogen and Fumitremorgin B by Penicillium piscarium Westling. AB - The tremorgenic mycotoxins verruculogen and fumitremorgin B were isolated from Penicillium piscarium Westling. The coexistence of these tremorgens in culture has previously been reported for one other unrelated fungal species, Aspergillus caespitosus Raper and Thom, and lends further support to the suggestion that the tremorgens have a common biosynthetic origin. PMID- 16345235 TI - Preparation and Activity of Immobilized Acetobacter suboxydans Cells. AB - The possibility of immobilization of Acetobacter suboxydans cells by entrapment within polyacrylamide gels or by intercellular cross-linking with glutaral-dehyde was investigated. The oxidation of unsubstituted alditols and aldose diethyl dithioacetals by the resultant modified cells has been achieved. PMID- 16345236 TI - Comparative Study of Two Methods for Detection of Clostridium perfringens in Ground Beef. AB - The tryptose-sulfite-cycloserine agar pour plate method was superior to selective enrichment in liquid sulfite medium for isolation of small numbers of Clostridium perfringens from frozen ground beef. PMID- 16345237 TI - Isolation of Atypical Candida albicans from the North Sea. AB - Isolates of Candida albicans with sparse filamentation and weak fermentation were isolated from the surface microlayer of the North Sea, but not from subsurface waters. Such atypical isolates may be misidentified by using normal taxonomic procedures. PMID- 16345238 TI - Simple procedure for large-scale assays for chick interferon. AB - Details are given of a simple and rapid method for the determination of chick interferon concentrations, using the Cooke microtiter system. The reliability of the result of this assay was examined by multiple determinations and by comparison with the results of another known assay procedure. A comparison of the relative time and cost of performing the two assays is also given. PMID- 16345239 TI - Nitrogen fixation (acetylene reduction) in a salt marsh amended with sewage sludge and organic carbon and nitrogen compounds. AB - Seasonal distribution of nitrogen fixation by Spartina alterniflora epiphytes and in surface and soil samples was investigated in a Georgia salt marsh which was amended with sewage sludge or with glucose and/or ammonium nitrate. There was no significant difference between the rates of fixation in the unamended and sewage sludge plots. Additional perturbation experiments suggested that nitrogen addition indirectly stimulates nitrogen fixation by enhancing Spartina production and root exudation. Glucose additions, on the other hand, suppressed nitrogen fixation on a long-term basis. It is suggested that the microbial population in the soil out-competed the plants for the available nitrogen and in turn suppressed plant production and possibly root exudation. A comparison of nitrogen fixation in clipped and unclipped Spartina plots substantiated the suggestion that root exudation probably supports nitrogen fixation. Fixation in the clipped plots was significantly lower (P < 0.05) than the rates in the unclipped plots. PMID- 16345240 TI - Multiple syringe inoculator for agar plates. AB - The design and operation of a manually operated multiple syringe inoculator was described. Either 9 or 21 inoculations of constant volume could be made simultaneously. Up to 100 plates could be inoculated in 15 min with excellent reproducibility. No contact occurred between the inoculating needles and the agar surface. Construction was simple and inexpensive, with minimal maintenance. PMID- 16345241 TI - Autoclavable dispenser for disposable pipette tips. AB - An autoclavable dispenser for plastic disposable pipette tips is described that allows aseptic removal of individual tips while maintaining the sterility of the remaining tips. PMID- 16345242 TI - General model of microbial growth and decomposition in aquatic ecosystems. AB - A model capable of simulating freely suspended and attached decomposers, particulate organic matter, labile and refractory dissolved organic matter, inorganic nitrogen, and phosphate in the open-water portion of lakes is presented. Examples are given showing the utility of the model when coupled to the whole-ecosystem model CLEANER. PMID- 16345243 TI - Indonesian tape ketan fermentation. AB - Indonesian tape ketan is a fermentation in which a mold, Amylomyces rouxii Calmette (Chlamydomucor oryzae Went and Prinsen Geerligs), in combination with one or more yeasts such as Endomycopsis burtonii converts steamed rice to a sweet sour, slightly alcoholic paste. A study was made to determine the biochemical changes that occur in the substrate during fermentation. It was found that the product was ready for consumption after fermentation at 30 degrees C for 36 to 48 h. A. rouxii used about 30% of the total rice solids, resulting in a crude protein of 12% in 96 h, whereas the combination of the mold with E. burtonii reduced total solids by 50% in 192 h, causing crude protein to increase to 16.5%. Soluble solids increased from 5 to about 67% in 36 h and decreased to 12% at 192 h with A. rouxii alone, whereas soluble solids fell to about 8% at 192 h in the fermentation with both the mold and the yeast. The mold, by itself, reduced the starch content of the rice from 78 to 10% in 48 h and to less than 2% in 144 h. The mold plus yeast reduced the starch content to about 18% in 48 h; however the "starch" content did not fall below 6% even at 192 h, presumably because the yeast was producing glycogen, which was determined along with the residual starch. With both the mold and the mold plus yeast fermentations, reducing sugars increased from less than 1% to approximately 5% in 24 h and reached maximum concentration, 16 to 17%, between 36 and 48 h. A. rouxii by itself produced a maximum of about 5.6% (vol/vol) ethanol at 96 h. The highest concentration of ethanol (8%, vol/vol) was produced by the mold plus E. burtonii at 144 h. The mold by itself reduced the starting pH from 6.3 to about 4.0 in 48 h. The combination of the mold and yeast reduced the pH to 4.1 in 144 h. The mold increased total acidity to approximately 6.2 meq of H per 100 ml, and the combination of the mold and yeast increased the total acidity to 7.8 meq of H per 100 ml in 192 h. At 48 h there was practically no difference in the volatile acidity (0.20) for the combined fermentation compared with 0.26 meq of H per 100 ml for the mold fermentation. The mold and at least one species of yeast were required to develop the rich aroma and flavor of typical Indonesian tape. PMID- 16345244 TI - Use of lipid composition and metabolism to examine structure and activity of estuarine detrital microflora. AB - Earlier studies have shown that the activity of the estuarine detrital microflora measured by various enzyme activities, muramic acid and adenosine 5'-triphosphate (ATP) content, heterotrophic potentials, and respiratory activities correlates with the incorporation of C and P into the microbial lipids. In this study, these lipids were reproducibly fractionated into neutral lipid, glycolipid, and phospholipid classes. Distinct differences between the active microflora of oak leaves, sweet gum leaves, and pine needles were evidenced both in the rate of lipid synthesis and in the proportions of neutral lipids, glycolipids, and phospholipids. Successional changes in the microflora of leaves incubated in a semitropical estuary, previously suggested by ATP-to-muramic acid ratios and scanning electron micrography, were reflected in changes in the proportions of C in major lipid classes when analyzed from the same type of detritus. Short incubation times with C gave lipid compositions rich in phospholipids that are typical for the faster-growing bacterial populations; longer incubation with C gave lipid compositions richer in neutral and glycolipids, more characteristic of slower-growing eukaryotes or morphologically more complex prokaryotes. The metabolism of the lipids of the estuarine detrital microflora was examined by a pulse-chase experiment with C. Glycolipids lost C at a rate equal to the loss of C of the slow component of muramic acid. Individual phospholipids lost C from their backbone glycerol esters at different rates. PMID- 16345245 TI - Sensitive enzymatic assay for glucose determination in natural waters. AB - A new enzymatic method for glucose determination is described. It allows measurement of glucose concentration as low as 10 M. Such sensitivity makes this method particularly appropriate for estimation of glucose in natural-water bodies, generally without prior concentration or extraction. The method is based on the reaction between glucose and adenosine 5'-triphosphate, catalyzed by hexokinase to form glucose-6-phosphate. The amount of adenosine 5'-triphosphate consumed in this reaction, which is directly proportional to the amount of glucose present in the sample, is measured by the luciferin-luciferase assay. The optimal conditions for glucose determination by this method have been defined as follows: 20 min of incubation at 30 degrees C, magnesium concentration of 10 M, and pH in the range of 7.5 to 10.5. The specificity of the assay to different carbohydrates has also been studied. Recovery of known amounts of glucose added to Lake Kinneret water was in the range of 80 to 114%. Application of this method is demonstrated in eight monthly profiles of the glucose content in Lake Kinneret. PMID- 16345246 TI - Preparation of specifically labeled C-(lignin)- and C-(cellulose)-lignocelluloses and their decomposition by the microflora of soil. AB - Microbial decomposition of lignocellulose in soil was studied using radioisotope techniques. Natural lignocelluloses containing C in either their lignin or cellulose (glucan) components were prepared by feeding plants l-[U C]phenylalanine or d-[U-C]glucose, respectively, through their cut stems. Detailed chemical and chromatographic characterization of labeled lignocelluloses from three hardwood and three softwood species showed that those labeled by the [C]glucose incorporation method contained specifically labeled cellulosic components, whereas those labeled by the [C]phenylalanine incorporation method contained specifically labeled lignin components. Microbial degradation of these differentially labeled lignocelluloses was followed by monitoring CO(2) evolution from selected soil samples incubated with known amounts of radiolabeled lignocelluloses. The lignin components of the six woods were shown to be decomposed in soil 4 to 10 times more slowly than their cellulosic components. These rates of mineralization were comparable to the generalized patterns previously reported in the literature. The present technique, however, was thought to be simpler, more sensitive, and less prone to interference than methods previously available. PMID- 16345247 TI - Baked-bean waste: a potential substrate for producing fungal amylases. AB - Baked-bean waste was found to be a favorable substrate for amylase production by Aspergillus foetidus NRRL 337. Under optimum conditions, the yields of alpha amylase (EC 3.2.1.1) and glucoamylase (EC 3.2.1.3) were 47 and 226 U, respectively, per ml of the waste fermented. PMID- 16345248 TI - Procedures involving liquid media for detection of bacterial contamination in breweries. AB - [This corrects the article on p. 247 in vol. 33.]. PMID- 16345249 TI - Occurrence of Escherichia coli in Wild Cottontail Rabbits. AB - [This corrects the article on p. 563 in vol. 33.]. PMID- 16345250 TI - pH Measurements of Agar Culture Media by Using a Flat-Bottom Electrode. AB - A new design, flat-bottom combination pH electrode was evaluated for utility in determining the pH of prepared agar media by surface contact of the electrode with the agar media. PMID- 16345251 TI - Weight increase of cotton fibers during swelling in alkali as a sensitive measure of cellulose degradation in ruminal fluid. AB - Weight increase of cotton fiber in an 18% NaOH solution, termed "alkali centrifuge" or "AC" value, was measured after incubation of either 1 g or 100 mg of the fiber in ruminal fluid. The AC response was a sensitive measure of cellulolytic activity. Thus, fiber incubated at 21 and 51 degrees C exhibited major AC increases even when direct weight losses of the unswollen fiber were less than 2%. Similarly, progressive additions of acetic acid to ruminal fluid progressively depressed both AC response and direct weight loss, but the former was still easily measurable when the latter was not. In tightly closed, completely filled vials with high ratio of ruminal fluid to sample, AC increased greatly and rapidly, i.e., in 6 h. This time could be further reduced to 2 h by overnight "preincubation" of the ruminal fluid with cotton fiber before starting the test incubation. Certain surfactants used to aid wetting of the fiber had a low but measurable potency in inhibiting cellulose digestion, but other surfactants were non-inhibitory. The AC response was maintained when ruminal fluid was diluted with an equal amount of McDougall's "artificial saliva" solution. PMID- 16345252 TI - Distribution of myxobacters in aquatic habitats of an alkaline bog. AB - Ten species of myxobacteria were identified from samples from an alkaline bog and adjacent soils. The frequency of occurrence and the diversity of species were highest at the margin of the bog and were lowest in the center and bottom of the bog lake. PMID- 16345253 TI - Small pet aquarium frogs as a source of salmonella. AB - [This corrects the article on p. 1027 in vol. 33.]. PMID- 16345254 TI - Structure, growth, and decomposition of laminated algal-bacterial mats in alkaline hot springs. AB - Laminated mats of unique character in siliceous alkaline hot springs of Yellowstone Park are formed predominantly by two organisms, a unicellular blue green alga, Synechococcus lividus, and a filamentous, gliding, photosynthetic bacterium, Chloroflexus aurantiacus. The mats can be divided approximately into two major zones: an upper, aerobic zone in which sufficient light penetrates for net photosynthesis, and a lower, anaerobic zone, where photosynthesis does not occur and decomposition is the dominant process. Growth of the mat was followed by marking the mat surface with silicon carbide particles. The motile Chloroflexus migrates vertically at night, due to positive aerotaxis, responding to reduced O(2) levels induced by dark respiration. The growth rates of mats were estimated at about 50 mum/day. Observations of a single mat at Octopus Spring showed that despite the rapid growth rate, the thickness of the mat remained essentially constant, and silicon carbide layers placed on the surface gradually moved to the bottom of the mat, showing that decomposition was taking place. There was a rapid initial rate of decomposition, with an apparent half-time of about 1 month, followed by a slower period of decomposition with a half-time of about 12 months. Within a year, complete decomposition of a mat of about 2-cm thickness can occur. Also, the region in which decomposition occurs is strictly anaerobic, showing that complete decomposition of organic matter from these organisms can occur in the absence of O(2). PMID- 16345255 TI - Comparative Statistical Study of the Cornwall, LKB, and Micromedic Dispensers. AB - The working precisions of the 5.0-ml-capacity Cornwall, the LKB 2075, and the Micromedic model 25000 dispensers were compared, and the bias of the LKB dispenser was contrasted with that of the Micromedic dispenser. Three technologists used six Cornwall dispensers, three LKB units with four different pumps, and three Micromedic units with six different pumps to dispense (in 1.0-ml amounts) sucrose solution adjusted to match the normal viscosity of serum. Under these conditions, the LKB dispenser was approximately 30% more precise than the Micromedic dispenser, which was approximately four times as precise as the Cornwall dispenser. Although the left pump site of the Micromedic was slightly more precise than the LKB, overall the Micromedic was less precise than the LKB. Moreover, the LKB was easier to use than the Micromedic. PMID- 16345256 TI - Fungal Growth on C1 Compounds: Quantitative Aspects of Growth of a Methanol Utilizing Strain of Trichoderma lignorum in Batch Culture. AB - [This corrects the article on p. 758 in vol. 33.]. PMID- 16345257 TI - Problems of the acetylene reduction technique applied to water-saturated paddy soils. AB - The acetylene reduction assay for the measurement of N(2) fixation in a water saturated paddy soil is limited by the slow diffusion of acetylene and ethylene. In laboratory incubation tests, vigorous shaking after the assay period is needed to release ethylene into the gas within the assay vials. Shaking prior to the incubation is also effective for dissolving acetylene in the water-saturated soil. However, a water-saturated soil depth of less than 10 mm during incubation is recommended. In field assays, some amounts of ethylene remain in the water saturated soil phase of the acetylene reduction assay chamber, but stirring the water-saturated soil before sampling reduces the amount of ethylene remaining in soil. Evidence of a downward movement of acetylene and an upward movement of ethylene through rice plants was obtained. Because of the rapid transfer of acetylene to rice plant roots, an in situ acetylene reduction assay covering a rice hill is likely to detect nitrogen fixation in the proximity of roots where acetylene is easily accessible. Acetylene introduction to the water-saturated soil phase prior to assay did not greatly increase the acetylene reduction rate. Carbon dioxide enrichment in the assay chamber did not enhance nitrogen fixation in a paddy including rice and algae during a 1-day cycle. PMID- 16345258 TI - Dialysis continuous process for ammonium-lactate fermentation of whey: mathematical model and computer simulation. AB - A mathematical model was developed to describe a dialysis process for the continuous fermentation of whey lactose to lactic acid, with neutralization to a constant pH by ammonia. In the process, whey of a relatively high concentration is fed into the fermentor circuit at a relatively low rate so that the residual concentration of lactose is low. The fermentor effluent contains ammonium lactate, bacterial cells, and residual whey solids and could be used as a nitrogen-enriched feedstuff for ruminant animals. Only water is fed into the dialysate circuit at a relatively high rate. The dialysate effluent contains purified ammonium lactate and could be converted to lactic acid and ammonium sulfate for industry. The fermentation was specifically modeled as a set of equations representing material balances and rate relationships in the two circuits. Dialysis continuous fermentations, in general, were modeled by combining these equations and by using dimensionless parameters. The generalized model was then solved for the steady state and used to simulate the specific fermentation on a digital computer. The results showed the effects of various material and operational and kinetic parameters on the process and predicted that it could be operated efficiently. PMID- 16345259 TI - Dialysis continuous process for ammonium-lactate fermentation of whey: experimental tests. AB - Laboratory experiments were conducted to validate theoretical predictions describing a dialysis continuous process for the fermentation of whey lactose to ammonium lactate, in which the fermentor contents are poised at a constant pH by adding ammonia solution and dialyzed through a membrane against water. Dried sweet-cheese whey was rehydrated to contain 230 mg of lactose per ml, supplemented with 8 mg of yeast extract per ml, charged into a 5-liter fermentor without sterilization, adjusted in pH (5.3) and temperature (44 degrees C), and inoculated with Lactobacillus bulgaricus. The fermentor and dialysate circuits were connected, and steady-state conditions were established. A series of such conditions was managed nonaseptically for 94 days to study the process and to demonstrate efficiency and productivity. As time progressed, the fermentation remained homofermentative and increased in conversion efficiency, although membrane fouling necessitated dialyzer cleaning about every 4 weeks. With a retention time of 19 h, 97% of the substrate was converted into products. Relative to nondialysis continuous or batch processes for the fermentation, the dialysis continuous process enabled the use of more concentrated substrate, was more efficient in the rate of substrate conversion, and additionally produced a second effluent of less concentrated but purer ammonium lactate. PMID- 16345260 TI - Factors Affecting Infection of Scenedesmus obliquus by a Chytridium sp. in Sewage Oxidation Ponds. AB - In a high-rate oxidation pond, 0.1 to 1.0% of the algal population of Scenedesmus obliquus was found to be infected by a chytrid. When suitable conditions developed, these infections burst into massive epidemics that killed most of the algae. The major factors triggering massive infections were optimal oxygen concentration and low concentrations of potassium and magnesium cations. The fungicide Benomyl was effective in preventing infection at a concentration of 1 mg/liter. PMID- 16345261 TI - Ruminococcus flavefaciens Cell Coat and Adhesion to Cotton Cellulose and to Cell Walls in Leaves of Perennial Ryegrass (Lolium perenne). AB - Ruminococcus flavefaciens was shown to possess a prominent glycoprotein coat, which contained rhamnose, glucose, and galactose as its principal carbohydrates. Periodate-reactive carbohydrate occurred as a surface layer of the coat. The ruminococci adhered strongly by means of this coat to cotton cellulose and to cell walls in leaf sections of Lolium perenne L. (perennial ryegrass). The coat was diffuse at the point of contact so that the bacterial cell wall was in close contact with the substrate. Adhesion was influenced by the availability of damaged plant cell walls and by the cell wall type and occurred most rapidly to cell walls of the epidermis and sclerenchyma, followed by the phloem and mesophyll. Plaques of bacteria with filamentous coat extensions developed on all these tissues. The bacteria did not readily adhere to the walls of the bundle sheath cells or metaxylem or protoxylem vessels and did not adhere to the cuticle or chloroplasts. The epidermal and phloem cell walls were more rapidly digested than the walls of other cell types. PMID- 16345262 TI - Effects of substrate biodegradability on the mass and activity of the associated estuarine microbiota. AB - Multiple biochemical assays of microbial mass and activities were applied to the estuarine detrital microbiota colonizing morphologically similar polyvinyl chloride needles and needles from slash pine (Pinus elliottii). Biodegradable pine needles consistently showed 2- to 10-fold higher values of extractable adenosine 5'-triphosphate, rates of oxygen utilization, activities of alkaline phosphatase and phosphodiesterase, and the mucopeptide cell wall component muramic acid than did the polyvinyl chloride needles, during a 14-week incubation in a semitropical estuary. The higher activities by the microbiota of the biodegradable substrate correlated with estimates of the microbial density from scanning electron microscopy. The microbial community associated with the nondegradable substrate showed minimal activity of beta-d-galactosidase, beta-d glucosidase, and alpha-d-mannosidase in contrast to the biota of the degradable substrate, which showed 10- to 100-fold higher activities of these glycoesterases. These enzymes logically could be involved in catabolism of the carbohydrate polymers of the detritus. Assuming equivalent rates of predation, a surface that is also a utilizable substrate supports a three- to fivefold more active microbial population. PMID- 16345263 TI - Identification of beijerinckia in the high arctic (devon island, northwest territories). AB - Although generally considered, with few exceptions, to be restricted to the acidic, tropical soils of the southern hemisphere, Beijerinckia species, resembling B. indica, were found at three sites on Devon Island (75 degrees 33'N, 84 degrees 40'W) in the Canadian Arctic. PMID- 16345264 TI - Persistence of Rhizobium japonicum on the Soybean Seed Coat Under Controlled Temperature and Humidity. AB - When Rhizobium japonicum strain 61A68 was added to surface-sterilized soybean (Glycine max) seed along with 12 different coating materials, a definite effect of temperature upon survival was observed both with and without coating materials. At a storage temperature of 15 degrees C and 50 +/- 5% relative humidity, from 0.9 to 14.1% of the original inoculum survived for 3 weeks. At 22.5 degrees C, from 0.5 to 7.2% of the original inoculum survived. At 30 degrees C, from 0.1 to 1.6% of the original inoculum survived. The data indicated that extremely large numbers of R. japonicum would have to be added to the seed to have numbers adequate for nodulation survive for 3 weeks of storage at ordinary temperatures. PMID- 16345265 TI - Effect of chlorine substitution on the biodegradability of polychlorinated biphenyls. AB - Thirty-one isomers of polychlorinated biphenyl (PCB) were examined for biodegradability by two species of Alcaligenes and Acinetobacter. The following relationships between chlorine substitution and biodegradability of PCBs were observed. (i) Degradation decreased as chlorine substitution increased. PCB isomers containing more than four chlorines were less susceptible to degradation. (ii) PCBs containing two chlorines on either the ortho position of a single ring (i.e., 2,6-) or on both rings (i.e., 2,2'-) showed very poor degradability. (iii) PCBs containing all chlorine atoms on only a single ring were generally degraded faster than when the same number of chlorines were substituted on both rings. (iv) Preferential ring fission of the molecules occurred with nonchlorinated or lesser chlorinated rings. (v) The formation and accumulation of a yellow intermediate was always observed in 4'-chloro-substituted PCBs. (vi) Significant differences between the two organisms with respect to degradability were not observed except for 2,4,6-trichlorobiphenyl. PMID- 16345266 TI - Metabolism of Di- and Mono-n-Butyl Phthalate by Soil Bacteria. AB - Di-n-butyl phthalate and other dialkyl phthalates are used as carbon sources by three Nocardia sp. isolates; mono-n-butyl phthalate is used as a carbon source by an Arthrobacter sp. isolate and a Pseudomonas sp. isolate. The compounds were metabolized in these organisms by hydrolysis to the corresponding monoesters and free phthalic acid. Phthalic acid was then metabolized via protocatechuic acid by 3,4-dioxygenative ring cleavage. PMID- 16345267 TI - Recovery of Poly-beta-Hydroxybutyrate from Estuarine Microflora. AB - Poly-beta-hydroxybutyrate (PHB) is a uniquely procaryotic endogenous storage polymer whose metabolism has been shown to reflect environmental perturbations in laboratory monocultures. When hydrolyzed for 45 min in 5% sodium hypochlorite, PHB can be isolated from estuarine detrital microflora in high yield and purified free from non-PHB microbial components. Lyophilization of frozen estuarine samples shortens the exposure time to NaOCl necessary for maximal recovery. Lyophilized samples of hardwood leaves, Vallisneria, and the aerobic upper millimeter of estuarine muds yielded PHB. The efficiency of incorporation of sodium [1-C]acetate into PHB is very high and is stimulated by aeration. PHB was not recovered from the anaerobic portions of sediments unless they were aerated for a short time. Levels of PHB in the detrital microbial community do not correlate with the microbial biomass as measured by the extractible lipid phosphate, suggesting that PHB-like eucaryotic endogenous storage materials may more accurately reflect the metabolic status of the population than its biomass. PMID- 16345268 TI - Denitrification and ammonia formation in anaerobic coastal sediments. AB - Simultaneous determinations of nitrogen gas production, ammonia, and particulate organic nitrogen formation in the coastal sediments of Mangoku-Ura, Simoda Bay, and Tokyo Bay were made by using the N-label tracer method. The rate of nitrogen gas production in the sediment surface layer was about 10 mug atom of N per g per h, irrespective of the location of the sediments examined. [N]ammonia and particulate organic nitrogen accounted for 20 to 70% of the three products, and after several hours of incubation, the major fraction of nondenitrified N in Mangoku-Ura and Simoda Bay sediments was recovered as ammonia. In Tokyo Bay sediments, particulate organic nitrogen was produced at a greater rate than was ammonia. The reduction rate data suggest that the pathway of nitrate reduction to ammonia is important in coastal sediments. PMID- 16345269 TI - Capacity for denitrification and reduction of nitrate to ammonia in a coastal marine sediment. AB - The capacity for dissimilatory reduction of NO(3) to N(2) (N(2)O) and NH(4) was measured in NO(3)-amended marine sediment. Incubation with acetylene (7 x 10 atmospheres [normal]) caused accumulation of N(2)O in the sediment. The rate of N(2)O production equaled the rate of N(2) production in samples without acetylene. Complete inhibition of the reduction of N(2)O to N(2) suggests that the "acetylene blockage technique" is applicable to assays for denitrification in marine sediments. The capacity for reduction of NO(3) by denitrification decreased rapidly with depth in the sediment, whereas the capacity for reduction of NO(3) to NH(4) was significant also in deeper layers. The data suggested that the latter process may be equally as significant as denitrification in the turnover of NO(3) in marine sediments. PMID- 16345270 TI - Microbial transformation of polycyclic aromatic hydrocarbons in pristine and petroleum-contaminated sediments. AB - To determine rates of microbial transformation of polycyclic aromatic hydrocarbons (PAH) in freshwater sediments, C-labeled PAH were incubated with samples from both pristine and petroleum-contaminated streams. Evolved CO(2) was trapped in KOH, unaltered PAH and polar metabolic intermediate fractions were quantitated after sediment extraction and column chromatography, and bound cellular C was measured in sediment residues. Large fractions of C were incorporated into microbial cellular material; therefore, measurement of rates of CO(2) evolution alone would seriously underestimate transformation rates of [C]naphthalene and [C]anthracene. PAH compound turnover times in petroleum contaminated sediment increased from 7.1 h for naphthalene to 400 h for anthracene, 10,000 h for benz(a)anthracene, and more than 30,000 h for benz(a)pyrene. Turnover times in uncontaminated stream sediment were 10 to 400 times greater than in contaminated samples, while absolute rates of PAH transformation (micrograms of PAH per gram of sediment per hour) were 3,000 to 125,000 times greater in contaminated sediment. The data indicate that four- and five-ring PAH compounds, several of which are carcinogenic, may persist even in sediments that have received chronic PAH inputs and that support microbial populations capable of transforming two- and three-ring PAH compounds. PMID- 16345271 TI - Nutritional Requirements of Selenomonas ruminantium for Growth on Lactate, Glycerol, or Glucose. AB - The nutritional requirements of Selenomonas ruminantium HD4 for growth on glucose, glycerol, or lactate were investigated to clarify the results of previous studies and to relate the nutrition of the organism to its physiology. The organism required l-aspartate, CO(2), p-aminobenzoic acid, and biotin for growth on a lactate-salts medium that contained small amounts of dithiothreitol. Aspartate could be replaced by l-malate or fumarate but not by succinate or l asparagine. Requirements for growth with glycerol as an energy source were similar, except that aspartate was not required. With glucose as the energy source, neither aspartate nor p-aminobenzoic acid was required, but a requirement for volatile fatty acids, which could be met by n-valerate, was observed. CO(2) was required for growth on lactate or glycerol but not on glucose on complex media containing Trypticase and yeast extract. Sulfide could be used as the sole source of sulfur. PMID- 16345272 TI - Effects of polychlorinated biphenyls on growth and respiration of heterotrophic marine bacteria. AB - A number of marine bacterial isolates from both near-shore and open-ocean environments were tested for growth inhibition with exposure to low concentrations (1 to 100 mug/liter) of Aroclor 1254, a commercial mixture of polychlorinated biphenyls (PCBs). Of over 17 bacterial cultures tested, growth of only two open-ocean isolates, one a pseudomonad and the other a tetrad-forming coccus, was consistently inhibited by Aroclor at concentrations as low as 10 mug/liter (10 ppb). Growth inhibition was dose dependent over a concentration range of 10 to 100 mug/liter. The effects upon division rates and final cell yields of each bacterial isolate were greatest when PCBs were added to cultures with low cell densities or with lower specific growth rates. The pseudomonad also had reduced carotenoid levels and an altered filamentous morphology with Aroclor present at a concentration of 10 mug/liter, or more. The effects noted were reversible for at least 18 h after initial exposure. Concentrations of Aroclor in excess of those needed to stop growth had no detectable effect upon the respiration rate of cells of either culture. This suggests that the reduced division rates observed were not due to inability of PCB-treated cells to transport or catabolize the carbohydrate or amino acid substrates tested. PMID- 16345273 TI - Effects of polychlorinated biphenyls on macromolecular synthesis by a heterotrophic marine bacterium. AB - Growth rates and final cell yields of a polychlorinated biphenyl (PCB)-sensitive pseudomonad isolated from the open ocean were reduced in a dose-dependent manner by 10 to 100 mug of Aroclor 1254 per liter, a commercial mixture of PCB isomers added to its culture medium. Effects on growth rates were detected within 1 h (approximately one doubling time) of treatment. By 4 h posttreatment, the amounts of deoxyribonucleic acid and ribonucleic acid per cell in exponentially growing populations treated with sublethal doses of Aroclor were detectably lower than in appropriate controls. Corresponding cell protein values were slightly higher than in controls. Selective degradation of cell proteins or nucleic acids was not detected in cells whose growth was totally suppressed for 4 h by PCBs. Cells whose growth rate was inhibited 20 to 50% by Aroclor synthesized protein at normal rates for periods in excess of 5 h from the time the chlorinated hydrocarbons were added. In contrast, rates per cell of adenine uptake and adenine incorporation into deoxyribonucleic acid and total nucleic acids by the cells treated with PCBs were significantly lower than in control cells. Intracellular adenine pools of cells whose growth was inhibited to 20% of the control rate by PCBs were 30% smaller and appeared to require a longer interval to equilibrate than those of untreated cells. This may indicate impaired transport and/or efflux of this nucleic acid precursor through the membrane of affected cells. Inhibition of nucleic acid synthesis in this sensitive bacterium by PCBs could explain the observed inhibitory effects of the chlorinated hydrocarbons on its growth. PMID- 16345274 TI - Estimation of fermentation biomass concentration by measuring culture fluorescence. AB - The fluorescence of a fermentation culture was studied for its application as an estimator of biomass concentration. The measurement was obtained by irradiating the culture with ultraviolet light (366 nm) through a glass window and detecting fluorescent light at the window surface at 460 nm. It was estimated that over one half of the fluorescent material was intercellular reduced nicotinamide adenine dinucleotide, with the remainder being reduced nicotinamide adenine dinucleotide phosphate and other unidentified intercellular and extracellular fluorophores. The culture fluorescence was found to be a function of biomass concentration, together with environmental factors, which presumably act at the cellular metabolic level to modify intercellular reduced nicotinamide adenine dinucleotide pools (e.g., dissolved oxygen tension, energy substrate concentration, and inhibitors). When these environmental conditions were controlled, a linear relationship was obtained between the log of the biomass concentration and the log of the fluorescence. Under these conditions, this relationship has considerable potential as a method to provide real-time biomass concentration estimates for process control and optimization since the fluorescence data is obtained on line. When environmental conditions are variable, the fluorescence data may be a sensitive index of overall culture activity because of its dependence on intercellular reduced nicotinamide adenine dinucleotide reserves and metabolic rates. This index may provide information about the period of maximum specific productivity for a specific microbial product. PMID- 16345275 TI - Methane, carbon dioxide, and hydrogen sulfide production from the terminal methiol group of methionine by anaerobic lake sediments. AB - A significant portion of the sulfide in lake sediments may be derived from sulfur containing amino acids. Methionine degradation in Lake Mendota (Wisconsin) sediments was studied with gas chromatographic and radiotracer techniques. Temperature optimum and inhibitor studies showed that this process was biological. Methane thiol and dimethyl sulfide were produced in sediments when 1 mumol/ml unlabeled methionine was added. When chloroform (an inhibitor of one carbon metabolism) was added to the sediments, methane thiol, carbon disulfide, and n-propane thiol were produced, even when no methionine was added. When S labeled methionine was added to the sediments in tracer quantities (1.75 nmol/ml), labeled hydrogen sulfide was produced, and a roughly equal amount of label was incorporated into insoluble material. Methane and carbon dioxide were produced from [methyl-C]methionine. Evidence is given favoring methane thiol as an intermediate in the formation of methane, carbon dioxide, and hydrogen sulfide from the terminal methiol group of methionine. Methionine may be an important source of sulfide in lake sediments. PMID- 16345276 TI - Hydrocarbon biodegradation in hypersaline environments. AB - When mineral oil, hexadecane, and glutamate were added to natural samples of varying salinity (3.3 to 28.4%) from salt evaporation ponds and Great Salt Lake, Utah, rates of metabolism of these compounds decreased as salinity increased. Rate limitations did not appear to relate to low oxygen levels or to the availability of organic nutrients. Some oxidation of l-[U-C]glutamic acid occurred even at extreme salinities, whereas oxidation of [1-C]hexadecane was too low to be detected. Gas chromatographic examination of hexane-soluble components of tar samples from natural seeps at Rozel Point in Great Salt Lake demonstrated no evidence of biological oxidation of isoprenoid alkanes subject to degradation in normal environments. Some hexane-soluble components of the same tar were altered by incubation in a low-salinity enrichment culture inoculated with garden soil. Attempts to enrich for microorganisms in saline waters able to use mineral oil as a sole source of carbon and energy were successful below, but not above, about 20% salinity. This study strongly suggests a general reduction of metabolic rate at extreme salinities and raises doubt about the biodegradation of hydrocarbons in hypersaline environments. PMID- 16345277 TI - Rapid determination of bacteriological water quality by using limulus lysate. AB - The Limulus lysate assay was used to measure the endotoxin content in stream water and was found to reflect the degree of bacterial contamination as measured by coliform, enteric, gram-negative, and heterotrophic bacteria. The firm-clot method was found to be a less sensitive and reproducible technique for the detection of endotoxin than was the spectrophotometric modification of the Limulus lysate assay. Bound endotoxin, as determined by the spectrophotometric modification of the Limulus lysate assay, was found to be a better measure of the endotoxin associated with bacterial cells than was total endotoxin. PMID- 16345278 TI - Growth of heterotrophic bacteria and algal extracellular products in oligotrophic waters. AB - The unexpected observation of 200 to 400 coliform bacteria per 100 ml in an unpolluted pristine stream was studied within Grand Teton National Park, Wyo. The high numbers of waterborne bacteria occurred in mid- to late summer at a location where there was a coincidental bloom of an algal mat community. Periphyton samplers were used to measure the algal growth that coincided with the increase in number of bacteria. Laboratory studies followed the growth of various coliform bacteria in the supernatant obtained from a Chlorella culture isolated from the mat community. Mixed natural bacterial populations from the stream and pure cultures of water-isolated fecal and nonfecal coliforms increased by two to three orders of magnitude at 13 degrees C when grown in the algal supernatant. Radioactive algal products were obtained by feeding an axenic Chlorella culture C labeled bicarbonate under laboratory cultivation at 13 degrees C with illumination. Radioactive organic material from the algae became incorporated into the particulate fraction of pure cultures of coliform bacteria as they reproduced and was later released as they died. PMID- 16345279 TI - Comparison of light and electron microscopic determinations of the number of bacteria and algae in lake water. AB - Determinations of the number of microorganisms in lake water samples with the bright-field light microscope were performed using conventional counting chambers. Determinations with the fluorescence microscope were carried out after staining the organisms with acridine orange and filtering them onto Nuclepore filters. For transmission electron microscopy, a water sample was concentrated by centrifugation. The pellet was solidifed in agar, fixed, dehydrated, embedded in Epon, and cut into thin sections. The number and area of organism profiles per unit area of the sections were determined. The number of organisms per unit volume of the pellet was then calculated using stereological formulae. The corresponding number in the lake water was obtained from the ratio of volume of solidified pellet/volume of water sample. Control experiments with pure cultures of bacteria and algae showed good agreement between light and electron microscopic counts. This was also true for most lake water samples, but the electron microscopic preparations from some samples contained small vibrio-like bodies and ill-defined structures that made a precise comparison more difficult. Bacteria and small blue-green and green algae could not always be differentiated with the light microscope, but this was easily done by electron microscopy. Our results show that transmission electron microscopy can be used for checking light microscopic counts of microorganisms in lake water. PMID- 16345280 TI - Distribution of Methylomonas methanica and Methylosinus trichosporium in Cleveland Harbor as Determined by an Indirect Fluorescent Antibody-Membrane Filter Technique. AB - An indirect fluorescent antibody-membrane filter staining technique, which permitted the autecological study of Methylomonas methanica and Methylosinus trichosporium, was developed. This technique was used to assay the numbers of these two organisms in Cleveland Harbor. The concentrations of M. methanica and M. trichosporium were found to be inversely proportional to the sampling depth, with the highest cell counts observed in the sediments. M. methanica was observed at every sampling station, whereas M. trichosporium was found at only two of the stations. PMID- 16345281 TI - Method for adenosine 5'-triphosphate measurement on coke waste activated sludge. AB - Measurement of adenosine 5'-triphosphate (ATP) in coke waste activated sludge can provide a simple method for estimating the levels of viable microbes in the sludge. However, the presence of inhibitors such as phenol in the sludge interferes when the luciferin-luciferase method is used to measure ATP. These inhibiting substances can be removed from the sludge before extraction of ATP by washing the cells with dilute sodium dodecyl sulfate. PMID- 16345282 TI - Respiration correction for microbial heterotrophic activity assays that use tritium-labeled substrates. AB - Filtrates from microbial heterotrophic activity assay experiments in which tritiated glucose was used as a radioactively labeled substrate were collected, and their radioactivities were determined. These filtrates were subsequently evaporated to dryness to remove tritiated water generated by metabolism of the labeled glucose; the residue was suspended to original volume with distilled water, and the tritium levels were again assayed. In the water samples assayed, the amount of tritium label retained by the microorganism was about 75% of that respired. PMID- 16345283 TI - Macromolecule Synthesis of Escherichia coli BB at a Lower or Transient Growth State. AB - [This corrects the article on p. 751 in vol. 34.]. PMID- 16345284 TI - Double-Stranded Ribonucleic Acid in Viruses of Penicillium citrinum. AB - [This corrects the article on p. 811 in vol. 34.]. PMID- 16345285 TI - Effect of Lactose Concentration on the Efficiency of Plating of Bacteriophages on Streptococcus cremoris. AB - The efficiency of plating of phages derived by ultraviolet induction of, or by lytic growth on, certain strains of Streptococcus cremoris was found to vary by as much as 10 depending on the lactose concentration of the medium in which the indicator bacteria were grown and the length of time the stationary-phase indicator cultures were aged. This effect was noted only when the culture was used as an indicator for phages that had previously grown on an apparently unrelated strain of bacteria. Conditions of culturing and aging had no detectable effect upon the ability of a strain to serve as an indicator for phage that had previously been cultured on the same strain. These observations suggest the presence of some kind of physiologically labile restriction system in strains of S. cremoris. The implications of this finding for increasing the sensitivity of the host range test in determining phage susceptibility, whether from induced lysates, whey, or lytic phage stocks, are discussed. It is recommended that, for all such testing, the concentration of lactose in buffered media be increased to such levels as required to obtain a final pH similar to that of a freshly coagulated milk culture, namely, below pH 5.0. PMID- 16345286 TI - Improved enrichment and isolation procedures for obtaining pure cultures of beggiatoa. AB - Scoring agar surfaces with alginate swabs before placing washed filaments of Beggiatoa on the agar has greatly increased the rate at which single filaments move from contaminated areas. Numerous morphological types of pure cultures have been grown in organic media supplemented with either catalase or reducing agents. Aerated sewage was used as the enrichment source. PMID- 16345287 TI - Effects of Lysine Analogs on Penicillium chrysogenum. AB - [This corrects the article on p. 706 in vol. 34.]. PMID- 16345288 TI - Effect of the Spartina alterniflora Root-Rhizome System on Salt Marsh Soil Denitrifying Bacteria. AB - Nitrous oxide (N(2)O) reductase activity was used as an index of the denitrification potential in salt marsh soils. In a short Spartina alterniflora marsh, the seasonal distribution of N(2)O reductase activity indicated a causal relationship between S. alterniflora root-rhizome production and the denitrification potential of the soil system. The relationship was not discerned in samples from a tall S. alterniflora marsh. To further examine the in situ plant-denitrifier interaction in the short S. alterniflora marsh, plots with and without living S. alterniflora were established and analyzed for N(2)O reductase activity 5 and 18 months later. In the plots without living Spartina there was a significant reduction in the soil denitrification potential after 18 months, indicating that in the SS marsh the denitrifiers are tightly coupled to the seasonal production of below-ground Spartina macroorganic matter. In plots with intact Spartina, the soil denitrification potential was not altered by NH(4)NO(3) or glucose enrichment. However, in plots without living Spartina, there were significant changes in soil N(2)O reductase activity, thus indicating that the plants can serve as a "buffer" against this form of pulse perturbation. PMID- 16345289 TI - Evaluation of the accuracy and precision of enumerating aerobic heterotrophs in water samples by the spread plate method. AB - Factors associated with accuracy and precision in the enumeration of aquatic aerobic heterotrophs by the spread plate method were evaluated by using a nested analysis of variance experimental design. Variances associated with individual components of the spread plate procedure were isolated, and optimal replications of each step were allocated. A practical scheme for optimal allocation of resources is proposed, consisting of four subsamples and two plates per subsample and yielding a total variance decrease of 70% from a single-subsample, 10-plate series. Data transformation was, in general, unnecessary for intraexperiment or intrasample statistical analysis, whereas interexperiment or intersample comparisons may require transformation of data. Rapid changes in the numbers of organisms in stored water samples were observed that were not reproducible and did not follow detectable trends, with increases or decreases in counts occurring in samples regardless of whether they were stored at room temperature or refrigerated, or stored in plastic or glass containers. Rapid sample handling is strongly recommended to minimize variations in the microbial populations of samples for aquatic environments. PMID- 16345290 TI - Reduction of Lactic Acid, Nonprotein Nitrogen, and Ash in Lactic Acid Whey by Candida ingens Culture. AB - A simple, efficient procedure for removing lactic acid and for reducing nonprotein nitrogen and ash in lactic acid whey has been developed. The procedure consists of culturing Candida ingens on the whey. This organism could assimilate >98% of the lactic acid and approximately 40% of the nonprotein nitrogen. Ash reduction of up to 45% resulted from precipitation of calcium apatite due to the increase in pH from 4.4 to approximately 8.0 which occurred during growth of C. ingens. Improved fluxes during laboratory-scale ultrafiltration were obtained for the treated lactic acid whey. C. ingens treatment of lactic acid whey appears to facilitate processing of this material to a more useful product. PMID- 16345291 TI - Plasmids, Lactic Acid Production, and N-Acetyl-d-Glucosamine Fermentation in Lactobacillus helveticus subsp. jugurti. AB - Two lactobacillus strains, Lactobacillus helveticus subsp. jugurti S13-8 and L. helveticus subsp. jugurti S36-2, were examined for the presence of plasmids. Plasmids of 16.45, 13.03, and 11.83 kilobases (kb) were found in the first, low lactic acid-producing strain; their function is not presently known. A single plasmid species of 13.17 kb was found in the second, high lactic acid-producing strain. This plasmid was found to be associated with lactic acid production and N acetyl-d-glucosamine fermentation. PMID- 16345292 TI - Method for sampling beef carcasses. AB - An instrument resembling a surgical dermatome, for microbiological sampling of beef carcasses, is described. PMID- 16345293 TI - Particle nature, yeast strain, and temperature interactions on the fermentation rates of grape juice. AB - Yeast strains of Saccharomyces cerevisiae behave differently to additions of finely sized particles added to clear grape juice. The type of material has a bearing on the effectiveness of increasing the rate of sugar conversion. In comparsion with a control sample, stirring and then adding grape solids back to the juice are most effective in maximizing sugar conversion, followed by addition of bentonite, talc, and diatomaceous earth. The improved fermentation efficiency is greater for all additions compared with unstirred controls. The formation of yeast flocs is enhanced by all additions. This fact can be used to partially explain the more effective fermentation rates. These additions appear to be equally effective over temperatures covering the normal fermentation temperature range. PMID- 16345294 TI - Interaction of bacteriophage infection and low penicillin concentrations on the performance of yogurt cultures. AB - Bacteriophage infection of a mixed-strain Streptococcus thermophilus culture, one strain of which is phage sensitive and the other phage resistant, may induce lysis of both strains. Experiments were carried out with three different phage resistant strains. One such strain lysed in penicillin-free growth medium and another needed penicillin G (0.005 IU/ml) for lysis, while the third strain continued to grow in the presence of this concentration of antibiotic. Growth of the latter strain was inhibited when the medium contained a relatively high concentration of phage lysin. The different penicillin concentrations required to induce "lysis from without" of these phage-resistant strains correlated with their individual sensitivities to the antibiotic. The apparent relationship between the sensitivities of these strains to penicillin and to phage lysin could be explained by a difference in the degree of polymerization of the cell wall peptidoglycan. PMID- 16345295 TI - Method for correcting laboratory model deep-well disposal system data for hydrostatic pressure effects. AB - A pressure chamber for determining the effect of increased hydrostatic pressure on growth and metabolic activities of groundwater bacteria is described. The chamber was used to show that moderate increases in pressure (to about 100 atmospheres) result in increased growth of mixed cultures of industrial-injection well bacteria and in the more complete degradation of formate and nitrate by these bacteria, as compared with identical cultures at atmospheric pressure. PMID- 16345299 TI - Bacterial predator-prey interaction at low prey density. AB - A bacterial predator-prey interaction was studied using Bdellovibrio and bioluminescent prey bacteria. The attacking bdellovibrio causes decay of bioluminescence, which is correlated with bdellovibrio penetration into the prey. The behavior of the prey and predator populations over time was found to be well described by a Lotka-Volterra model. By using this model, the probability of bdellovibrio penetration after encountering a prey cell was found to be approximately 3.0%. The prey density required to give the bdellovibrios a 50% chance of survival was calculated to be at least 3.0 x 10 cells per ml, and the density required for population equilibria was calculated to be about 7 x 10 prey bacteria per ml. These values, not generally characteristic of natural habitats, suggest that the existence of Bdellovibrio in nature is limited to special ecological niches. PMID- 16345300 TI - Expression of rhizobial nitrogenase: influence of plant cell-conditioned medium. AB - Conditioned medium was obtained from suspension cultures of soybean (Glycine max L. Merrit) cells after incubating them for 4 to 8 days with rhizobia which were separated from the soybean cells by two dialysis bags, one within another. This conditioned medium from the plant cell side (PCM) of the two membranes was used to elicit and influence nitrogenase activity (acetylene reduction) in rhizobia. When conditions for obtaining PCM from the soybean cell suspension cultures were varied, it could be shown that freshly grown rhizobia were able to induce active compounds in the PCM. These compounds caused acetylene reduction activity in test rhizobia under conditions where control rhizobia, containing various substrates, showed little or no acetylene reduction activity. Rhizobia that were already capable of acetylene reduction could not induce such compounds in the PCM when this was included with test rhizobia. The PCM from soybean cultures was also found to aid the expression of nitrogenase activity in suspension cultures of rhizobia normally associated with either peas, lupins, broad beans, or clovers. This is the first communication indicating nitrogenase activity in freeliving cultures for various species of rhizobia. PMID- 16345301 TI - Nitrogen fixation (acetylene reduction) by epiphytes of freshwater macrophytes. AB - The involvement of epiphytic microorganisms in nitrogen fixation was investigated in a shallow freshwater pond near Ithaca, N.Y. The acetylene reduction technique was used to follow diel and seasonal cycles of nitrogen fixation by epiphytes of Myriophyllum spicatum. Acetylene-reducing activity was maximal between noon and 6 p.m., but substantial levels of activity relative to daytime rates continued through the night. Experiments with the seasonal course of activity showed a gradual decline during the autumn months and no activity in January or February. Activity commenced in May, with an abrupt increase to levels between 0.45 and 0.95 nmol of ethylene formed per mg (dry weight) of plant per h. Through most of the summer months, mean rates of acetylene reduction remained between 0.15 and 0.60 nmol/mg (dry weight) per h. It was calculated from diel and seasonal cycles that, in the pond areas studied, epiphytes were capable of adding from 7.5 to 12.5 mug of N per mg of plant per year to the pond. This amount is significant relative to the total amount of nitrogen incorporated into the plant. Blue-green algae (cyanobacteria), particularly Gloeotrichia, appeared to bear prime responsibility for nitrogen fixation, but photosynthetic bacteria of the genus Rhodopseudomonas were isolated from M. spicatum and shown to support high rates of acetylene reduction. PMID- 16345302 TI - Denitrification rates in a marine sediment as measured by the acetylene inhibition technique. AB - A method has been developed for measurement of denitrification activity in sediments by application of the acetylene inhibition technique. Acetylene saturated water was injected, at close intervals, into sediment cores which were then incubated for a few hours at the in situ temperature. Frozen segments of the cores were assayed for accumulation of N(2)O by a combined gas extraction and detection system. The segments were thawed under a stream of helium from which N(2)O (and other gases) was collected in a liquid N(2) trap, and the quantity of N(2)O was measured by gas chromatography. The maximum rate of denitrification in a coastal marine sediment was 35 nmol of N per cm of sediment per day at 2.5 degrees C, and the rate of denitrification for the total sediment was 0.99 nmol of N per m per day. PMID- 16345303 TI - Electron microscopy of the microbial populations present and their modes of attack on various cellulosic substrates undergoing digestion in the sheep rumen. AB - Cotton fibers and various cell wall preparations from grass leaves and from the feces of sheep fed on dried grass were placed in the sheep rumen in bags made from 5-mum-mesh nylon cloth. After periods of from 3 to 48 h, bags were removed, and the contents were fixed, embedded, sectioned, and stained for electron microscopy. Some of the bacteria present were seen to be closely associated with the cell walls, either tunneling within them or making very close contact. Evidence was obtained for differential digestion of cell walls and of the layers within them. Distinct differences were noticed between bacterial populations attacking the more susceptible wall types and those attacking feces cell walls and cotton fibers. Among the latter, the dominant form was a long, thin rod with a typical gramnegative cell wall structure, different from that described for Bacteroides succinogenes S85 or for Butyrivibrio fibrisolvens. PMID- 16345305 TI - Comparison of solid-phase radioimmunoassays for baculoviruses. AB - The sensitivity and cross-reaction of four solid-phase radioimmunoassays (RIA) for Trichoplusia ni nuclear polyhedrosis virus containing singly enveloped virions were investigated. The detection limits of each assay were as follows: Indirect RIA, 5 ng of dissolved polyhedron antigen; direct RIA, 50 ng; indirect sandwich RIA, 200 ng; and direct sandwich RIA, 300 ng. The indirect and indirect sandwich RIAs showed considerable cross-reaction with other baculovirus antigens, but the direct and direct sandwich RIAs showed cross-reaction with only one closely related baculovirus. When microtiter plates used for the solid phase were pretreated with bovine serum albumin, nonspecific binding of labeled antibodies was reduced to a minimum. Antibodies prepared by an immunoadsorption procedure showed greater specific binding than antibodies prepared by ammonium sulfate precipitation of the immunoglobulin fraction. Highly contaminated antigen could not be detected by the indirect RIA, but the direct sandwich RIA was unaffected by antigen contamination. Antigen making up 0.0025% (wt/wt) of a sample of bird droppings could be detected by the direct sandwich RIA. PMID- 16345304 TI - Dependence of marine bdellovibrios on potassium, calcium, and magnesium ions. AB - Marine bdellovibrios show a specific requirement for K, Ca, and Mg. Potassium is essential for high velocity and seems to be necessary for attachment of the free bdellovibrios. Calcium and magnesium are necessary for attachment and penetration. Magnesium also plays a role in maintaining the integrity of the bdelloplast. The adaptation of these bdellovibrios to the marine environment is manifested by their stringent cation requirements. PMID- 16345306 TI - Plaque assay of nuclear polyhedrosis viruses in cell culture. AB - The nuclear polyhedrosis virus of Autographa californica has been titrated in Spodoptera frugiperda cells by the plaque method, using a solid overlay which does not require either the use of modified culture medium or expensive purified agarose or the addition of culture medium as a liquid layer above the solid agarose. This assay is more sensitive than that using a viscous methyl cellulose overlay but less sensitive than the end-point dilution technique. Neither Trichoplusia ni nor Bombyx mori cells were satisfactory as indicators for the assay as described, since they failed to form a stable monolayer. Manduca sexta cells could be utilized for assay of A. californica nuclear polyhedrosis virus, but the sensitivity was lower than with S. frugiperda cells. PMID- 16345307 TI - Sugar level, fermentability, and acceptability of straw treated with different acids. AB - The sugar level, fermentability, and acceptability by voles of ryegrass straw treated with different acids were compared. Sugar release was highest with HCl, but acceptability was best with H(3)PO(4) treatment. Candida utilis grew about equally well on all of the treated samples, except the one treated with 0.5 N HCl. That sample supported no growth, possibly because the level of chloride or degradation products of monomeric sugars was too high. Fermentability and palatability of straw were highest when treated with a combination of 0.23 N HCl and 0.15 N H(3)PO(4). Fermentation with Aureobasidium pullulans further increased the palatability of the acid-treated straw. PMID- 16345308 TI - Growth of Chaetomium cellulolyticum on Alkali-Pretreated Hardwood Sawdust Solids and Pretreatment Liquor. AB - The treatment of a hardwood sawdust with 1% NaOH solution at 121 degrees C dissolved 19.7% of the dry matter, mainly hemicellulose and lignin. Fermentation of the treated solids by Chaetomium cellulolyticum for 48 h gave a product containing 12.5% crude protein (total N x 6.25) on a dry weight basis. The in vitro rumen digestibility of the 48-h fermentation product was 30%, compared to 24% for the alkali-treated but unfermented sawdust. Growth was independent of sawdust particle size in the range 40 to 100 mesh. Fermentation of the pretreatment liquor gave a product containing up to 50% crude protein (dry weight basis) with an in vitro rumen digestibility of 65 to 76%. Approximately 6.7 g of crude protein was obtained from the treated solids and 2.2 g from the pretreatment liquor per 100 g of sawdust treated. The product from the pretreatment liquor fermentation has potential as a high-protein animal feed supplement but could not be produced economically without an outlet for the relatively indigestible product from the solids fermentation. Growth on the pretreatment liquor was strongly pH dependent; there was a considerable increase in the lag phase when the pH was lowered from 7.5 to 5.2. This effect appears to be due to an inhibitor whose toxicity is reduced at high pH. PMID- 16345309 TI - Growth of "seeded" cellulolytic enrichment cultures on mesquite wood. AB - Eleven enrichment cultures were developed by a "seeded" enrichment culture technique, and one was developed by a simple enrichment technique. The seeded enrichment, the pure "seed," and the simple enrichment cultures were compared during growth on mesquite wood, cotton, carboxymethylcellulose, and cellobiose. All of the enrichment cultures were cellulolytic and exceeded the pure seed cultures in mesquite wood hydrolysis and/or viable cell count. Yeast extract improved, but was not essential for, growth of the seeded enrichment cultures on carboxymethylcellulose. Two of the seeded enrichment cultures, CAD5 and CAD11, grew best at 37 degrees C and pH 7.0 on mesquite wood. A 1.0% (wt/vol) wood concentration was optimum for their growth. PMID- 16345310 TI - Measurement and significance of specific activity in the heterotrophic bacteria of natural waters. AB - It is now possible to obtain accurate total counts of the bacteria of natural waters with the use of acridine orange staining and epifluorescence microscopy. This approach can be coupled to highly sensitive measurements of heterotrophic activity using radioisotopes. To accomplish this, three variations of a "specific activity index" are suggested, based on different approaches to measuring heterotrophic activity with radiolabeled organic solutes. The denominator of each index is the direct count of bacteria from a given natural sample. Three numerators are presented, each of which has been shown to vary directly with heterotrophic bacterial activity: V(max), turnover rate, and direct uptake (at high substrate concentrations). Each approach is illustrated with data from estuarine and coastal waters of northeastern Massachusetts. The data show major differences in specific activity that accompany such habitat differences as distances within or offshore from an estuary and vertical location in the water column. These and other data suggest that specific activity is a valid indicator of the physiological state and metabolic role of the bacteria. Some evidence is presented in support of the hypothesis that the natural bacteria are adapted to conditions of nutrient starvation by becoming "dormant," existing for an unknown period of time in a reversible physiological state that reflects the availability of organic nutrients. PMID- 16345311 TI - Substrate preferences in rumen bacteria: evidence of catabolite regulatory mechanisms. AB - Five species of rumen bacteria with overlapping substrate fermentative capabilities were tested for substrate preferences and evidence of catabolite regulatory mechanisms. All five bacteria showed evidence of some type of catabolite regulatory mechanism. In the six-substrate test system that was used, utilization of every substrate was inhibited by another substrate in at least one of the bacteria. Inhibited versus noninhibited substrate data suggest that the five bacteria have different strategies of substrate utilization and thus occupy separate niches in the rumen. The significance of these observations to understanding the rumen ecosystem is discussed. PMID- 16345312 TI - Kinetic parameters of the conversion of methane precursors to methane in a hypereutrophic lake sediment. AB - The kinetic parameters K(m), V(max), T(t) (turnover time), and v (natural velocity) were determined for H(2) and acetate conversion to methane by Wintergreen Lake sediment, using short-term (a few hours) methods and incubation temperatures of 10 to 14 degrees C. Estimates of the Michaelis-Menten constant, K(m), for both the consumption of hydrogen and the conversion of hydrogen to methane by sediment microflora averaged about 0.024 mumol g of dry sediment. The maximal velocity, V(max), averaged 4.8 mumol of H(2) g h for hydrogen consumption and 0.64 mumol of CH(4) g h for the conversion of hydrogen to methane during the winter. Estimated natural rates of hydrogen consumption and hydrogen conversion to methane could be calculated from the Michaelis-Menten equation and estimates of K(m), V(max), and the in situ dissolved-hydrogen concentration. These results indicate that methane may not be the only fate of hydrogen in the sediment. Among several potential hydrogen donors tested, only formate stimulated the rate of sediment methanogenesis. Formate conversion to methane was so rapid that an accurate estimate of kinetic parameters was not possible. Kinetic experiments using [2-C]acetate and sediments collected in the summer indicated that acetate was being converted to methane at or near the maximal rate. A minimum natural rate of acetate conversion to methane was estimated to be about 110 nmol of CH(4) g h, which was 66% of the V(max) (163 nmol of CH(4) g h). A 15-min preincubation of sediment with 5.0 x 10 atm of hydrogen had a pronounced effect on the kinetic parameters for the conversion of acetate to methane. The acetate pool size, expressed as the term K(m) + S(n) (S(n) is in situ substrate concentration), decreased by 37% and T(t) decreased by 43%. The V(max) remained relatively constant. A preincubation with hydrogen also caused a 37% decrease in the amount of labeled carbon dioxide produced from the metabolism of [U-C]valine by sediment heterotrophs. PMID- 16345313 TI - Occurrence and ecological significance of GTP in the ocean and in microbial cells. AB - A comparison between the ATP concentrations based on peak height light emission values (0 to 3 s) and integrated light flux determinations (15 to 75 s) for a variety of seawater samples revealed that the integrated method of light detection consistently yielded higher ATP concentrations, ranging from 1.38 to 2.35 times larger than the corresponding peak ATP values. A significant correlation (r = 0.923) was observed for a plot of DeltaADP (i.e., integrated ATP - peak ATP) versus GTP + UTP, suggesting that the analytical interference on the ATP assay was the result of the presence of non-adenine nucleotide triphosphates. Size-fractionation studies revealed an enrichment of the non-adenine nucleotide triphosphates, relative to ATP, in the smallest size fraction analyzed (<10 mum). Investigations were conducted with 20 species of unicellular marine algae to determine their intracellular nucleotide concentrations, and these determinations were compared to the levels measured in lab cultures of the marine bacterium Serratia marinorubra. These results indicated that the intracellular GTP/ATP ratios in S. marinorubra increase in direct proportion to the rate of cell growth, and that the GTP/ATP ratios in bacteria are much greater than in growing algae, presumably due to the differences in rates of cellular biosynthesis. It is concluded that quantitative determinations of GTP/ATP ratios in environmental sample extracts may be useful for measuring microbial growth. PMID- 16345314 TI - Stabilization of Lactose Metabolism in Streptococcus lactis C2. AB - The integration of the lactose plasmid from lactic streptococci into the host chromosome could stabilize this trait for dairy fermentations. Sixty lactose positive (Lac) transductants of lactose- and proteinase-negative (Lac Prt) LM0220 were induced for temperature phage by UV irradiation or mitomycin C. Four of the transductants, designated KB18, KB21, KB54, and KB58, yielded lysates demonstrating less than one Lac transductant per 0.2 ml of phage lysate. Successive transferring in the presence of acriflavine did not yield Lac segregants from KB18, KB21, KB54, or KB58, whereas Streptococcus lactis C2 (parent culture) and three other Lac transductants showed 12 to 88% conversion from Lac to Lac within 6 to 10 repetitive transfers. When grown in continuous culture, KB21 did not show any Lac variants in 168 h, while S. lactis C2 had 96% conversion from Lac to Lac in 144 h. Agarose gel electrophoresis of plasmid DNA isolated from KB18, KB21, KB54, and KB58 revealed that the lactose plasmid, pLM2103, normally present in Lac transductants, was missing. This suggested integration of the transferred lactose plasmid into the chromosome. In contrast to phage lysates induced from S. lactis C2, which exhibited an exponential decrease in the number of Lac transductants after exposure to small doses of UV irradiation, the transduction frequency for lactose metabolism was stimulated by UV irradiation of lysates from KB58. The latter indicated chromosomal linkage for lac and that integration of the lactose genes plasmid into the chromosome had occurred. PMID- 16345315 TI - Rumen protozoal degradation of structurally intact forage tissues. AB - The association with and digestion of intact leaf sections of cool- and warm season grasses by cattle rumen protozoa were investigated by light and scanning electron microscopy and by in vitro dry matter disappearance studies. Within extensively degraded areas of mesophyll tissue in cool-season forages, almost all protozoa were Epidinium ecaudatum form caudatum, with maximum numbers at 4 to 10 h of incubation. However, few protozoa were found inside warm-season forage leaves. In in vitro dry matter disappearance studies of a series of incubations with and without 1.6 mg of streptomycin per ml, which inhibited the cellulolytic activity of the bacteria, and in comparison with uninoculated controls, rumen protozoa degraded 11.0 and 3.7 percentage units of orchardgrass and bermuda grass, respectively. Scanning electron microscopy showed that the tissues degraded in orchardgrass consisted of large amounts of mesophyll and portions of the parenchyma bundle sheath and epidermis; no tissue loss due to the protozoa was observed in bermudagrass. The relationship of these observations to forage digestion is discussed. PMID- 16345316 TI - Thermophilic iron-oxidizing bacteria found in copper leaching dumps. AB - Rod-shaped bacteria capable of oxidizing ferrous iron at 55 degrees C were cultured from samples of a copper mine leach dump. Yeast extract or cysteine was required by these Thiobacillus-like bacteria for growth, using ferrous iron as an energy source. PMID- 16345317 TI - Medium for the enumeration and isolation of bacteria from a Swine waste digester. AB - A habitat-simulating medium was developed for the enumeration and isolation of bacteria from a swine waste digester. A roll tube medium with growth factors for strict anaerobes from previously studied anaerobic ecosystems was used to evaluate the effects of deletion, addition, or level of digester fluid, digester fluid treated with acid or base, rumen fluid, fecal extract, anaerobic pit extract, tissue extract, carbohydrates, peptones, short-chain fatty acids, minerals, vitamins, N and P sources, reducing and solidifying agents, buffers, and gases on colony counts. Decreasing the agar concentration from 2.5 to 1.0% increased the counts twofold. Blending increased the counts 1.7-fold. With a medium (174) containing digester fluid, peptones, minerals, cysteine, sodium carbonate, and agar, colony counts were 60% of the microscopic count and improved yields 2.5 to 20 times those obtained with media previously used for digesters or developed for other anaerobic ecosystems. Colony counts continued to increase for up to 4 weeks of incubation. Medium 174 permits the enumeration of total, methanogenic, and, with deletion of reducing agent, aerotolerant bacteria. The results suggest that the predominant bacteria grow slowly and have requirements different from those of bacteria from other ecosystems. PMID- 16345318 TI - Effect of temperature on blue-green algae (cyanobacteria) in lake mendota. AB - The temperature optimum for photosynthesis of natural populations of blue-green algae (cyanobacteria) from Lake Mendota was determined during the period of June to November 1976. In the spring, when temperatures ranged from 0 to 20 degrees C, there were insignificant amounts of blue-green algae in the lake (less than 1% of the biomass). During the summer and fall, when the dominant phytoplankton was blue-green algae, the optimum temperature for photosynthesis was usually between 20 and 30 degrees C, whereas the environmental temperatures during this period ranged from 24 degrees C in August to 12 degrees C in November. In general, the optimum temperature for photosynthesis was higher than the environmental temperature. More importantly, significant photosynthesis also occurred at low temperature in these samples, which suggests that the low temperature alone is not responsible for the absence of blue-green algae in Lake Mendota during the spring. Temperature optima for growth and photosynthesis of laboratory cultures of the three dominant blue-green algae in Lake Mendota were determined. The responses of the two parameters to changes in temperature were similar; thus, photosynthesis appears to be a valid index of growth. However, there was little photosynthesis by laboratory cultures at low temperatures, in contrast to the natural samples. Evidence for an interaction between temperature and low light intensities in their effect on photosynthesis of natural samples is presented. PMID- 16345319 TI - Diversity in the ammonia-oxidizing nitrifier population of a soil. AB - Multiple genera of ammonia-oxidizing chemoautotrophic nitrifiers in a soil were detected, isolated, and studied by means of modified most-probable-number (MPN) techniques. The soil examined was a Waukegon silt loam treated with ammonium nitrate or sewage effluent. The genera Nitrosomonas and Nitrosospira were found to occur more commonly than the genus Nitrosolobus. Three different MPN media gave approximately the same overall ammonia oxidizer counts within statistical error after prolonged incubation but differed markedly in ratios of Nitrosomonas to Nitrosospira. Selectivity and counting efficiency of MPN media were studied by observing the growth response of representative pure cultures isolated from the soil. Selectivity was evident in each medium with respect to all strains tested, and the media differed greatly in incubation times required to obtain maximum counts. PMID- 16345320 TI - Serological diversity within a terrestrial ammonia-oxidizing population. AB - Fluorescent antibodies (FAs) prepared against 16 ammonia-oxidizing nitrifying bacteria were examined as to cross-reactivity in heterologous FA staining tests. Virtually all cross-reactions to the seven Nitrosomonas FAs were confined to the Nitrosomonas isolates. The five Nitrosospira, three Nitrosolobus, and one Nitrosovibrio FAs stained isolates only within their respective genera. Within each genus shared antigens were more common among Nitrosomonas and Nitrosolobus than among Nitrosospira isolates. Isolates obtained from a single soil were stained to examine the effectiveness of the suite of FAs for study of a given ammonia-oxidizing population. Of the seven Nitrosomonas isolates, five were stained effectively through the use of four FAs; six of twelve Nitrosospira isolates were identifiable through use of four different FAs. The one Nitrosolobus isolate was stained well only by one (its homologous) FA. PMID- 16345321 TI - Production and ecological significance of yeast cell wall-degrading enzymes from oerskovia. AB - Motile actinomycetes capable of degrading walls of viable yeast cells were isolated from soil and identified as Oerskovia xanthineolytica. A lytic assay based on susceptibility of enzyme-treated cells to osmotic shock was developed, and 10 of 15 strains of O. xanthineolytica, Oerskovia turbata, and nonmotile Oerskovia- like organisms from other collections were found to possess yeast lytic activities. All lytic strains produced laminaranase and alpha-mannanase, but the amounts, determined by reducing group assays, were not proportional to the observed lytic activities. The Oerskovia isolates demonstrated chemotactic, predatory activity against various yeast strains and killed yeasts in mixed cultures. Of 15 carbon sources tested for production of lytic enzyme, purified yeast cell walls elicited the highest activity. Glucose repressed enzyme production and caused cells to remain in the microfilamentous and motile rod stages of the Oerskovia cell cycle. Crude lytic activity was optimal at pH 5.6 to 7.0 and inactivated by heating for 6 min at 50 degrees C. Partial purification by isoelectric focusing showed that all lytic activity was associated with four beta (1-->3)-glucanases. The absence of protein disulfide reductase, N-acetyl-beta-d hexosaminidase, and phosphomannanase in crude preparations indicated that the principal enzyme responsible for yeast wall lysis was a beta-(1-->3)-glucanase that produced relatively little reducing sugar from yeast glucan. PMID- 16345322 TI - Cellulolytic Enzyme System of Thermoactinomyces sp. Grown on Microcrystalline Cellulose. AB - The carboxymethyl-cellulase and Avicelase activities of Thermoactinomyces sp., strain YX, were produced simultaneously with cell growth. Throughout the growth phase these activities were primarily extracellular, with up to 50% adsorbed to residual cellulosic substrate at any one time. On the other hand, the beta glucosidase activity was associated with the culture solids throughout the entire fermentation and appears to be intracellular. Preparative isoelectric focusing of the culture filtrate, in the pH range 3 to 5, separated three major fractions with cellulolytic activities towards both carboxymethyl-cellulose and Avicel. Based on the carboxymethyl-cellulase and Avicelase activities of these separated fractions, it was not possible to discriminate between endo- and exoglucanases produced by Thermoactinomyces sp. Analytical isoelectric focusing of culture filtrates obtained throughout the growth phase of Thermoactinomyces indicated that all the extracellular cellulolytic enzymes are produced and released into the culture filtrate simultaneously, with no evidence of a sequential appearance of the various enzymes or isoenzymes. PMID- 16345323 TI - Extracellular Accumulation of a New Amino Acid, O-2-Hydroxypropylhomoserine, from 1,2-Propanediol by Flavobacterium rigense. AB - [This corrects the article on p. 1046 in vol. 35.]. PMID- 16345324 TI - Changes in proportions of acetate and carbon dioxide used as methane precursors during the anaerobic digestion of bovine waste. AB - [This corrects the article on p. 651 in vol. 35.]. PMID- 16345325 TI - Degradation of tyrosine in anaerobically stored piggery wastes and in pig feces. AB - Radioactively labeled compounds that might be intermediates in the anaerobic degradation of tyrosine were added to pig feces and to stored piggery wastes. Changes in the compounds were followed by using thin-layer and gas chromatography. In feces, p-cresol and 3-phenylpropionic acid were the end products of tyrosine metabolism; in anaerobically stored mixed wastes, phenol, p cresol, and minor quantities of phenylpropionic acid were formed. Schemes were proposed for the degradation of tyrosine in pig feces and in mixed wastes. PMID- 16345326 TI - Factors affecting the activity of cellulases isolated from the rumen digesta of sheep. AB - Sodium phosphate buffer was used to extract cellulases from the plant solids fraction of rumen contents. The mixed cellulase preparation had maximal activity at pH 6.9 and 49 degrees C. The V(max) and the apparent K(m) for wheaten hay cellulose were 19.8 glucose units/min and 6.35 mg/ml, respectively, and for microcrystalline cellulose (Sigmacell) at the same enzyme concentration, they were 33 glucose units/min and 27.5 mg/ml, respectively. For these assays a glucose unit was defined as nanomoles of glucose plus twice the nanomoles of cellobiose. Consideration of thermodynamic and kinetic data suggested that the hydrolysis of a relatively labile arabino-xylan comprising 3% of the wheaten hay cellulose was dependent on prior removal of the protecting beta-1,4-glucose chains at the outer surface of the cellulose preparation. Sequential removal of structural polysaccharides from the plant cell wall rendered the latter more susceptible to cellulase activity. Cellulase activity was stimulated by increasing the concentration of phosphate from 5 to 50 mM. The stimulation was magnified in the presence of cell-free rumen fluid. Cellulase activity was not stimulated by calcium, magnesium, iron, zinc, manganese, copper, or cobalt ions and was unaffected by the chelators ethylenediaminetetraacetic acid and ethyleneglycol-bis (beta-aminoethyl ether)-N,N'-tetraacetic acid. O phenanthroline inhibited activity by 30 to 50%, but this may have been due to nonchelate properties. Anaerobic conditions or thiol protective agents were not essential for either the activity or stability of the cellulases during assay. An ultrafiltrable inhibitor of cellulase activity was detected in cell-free rumen fluid. PMID- 16345327 TI - Selective Growth Inhibition of Sphaerotilus natans and Beggiatoa sp. by Nucleosides. AB - 9-beta-d-Arabinofuranosyladenine (ara-A) has been found to specifically inhibit the growth of Sphaerotilus natans and Beggiatoa sp. at a low concentration (0.78 mug/ml). The nucleoside had no antimicrobial activity against various microorganisms other than Candida albicans at 1,000 mug/ml. 3'-Deoxyadenosine, 2' deoxyadenosine, formycin, and some derivatives of ara-A also showed inhibitory activity against Sphaerotilus natans. The growth of Beggiatoa sp. was also inhibited by 9-beta-arabinofuranosylhypoxanthine, 3'-deoxyadenosine, 2' deoxyadenosine, formycin, toyocamycin, tubercidin, and some derivatives of ara-A. ara-A was quite stable in water and had no harmful effect on fish at 200 mug/ml. The possible uses of ara-A and some nucleosides in controlling the proliferation of S. natans and Beggiatoa sp. in the environment are discussed. PMID- 16345328 TI - Microbiological and physical properties of salt marsh and microecosystem sediments. AB - Sediments from natural marsh and four microecosystem (MES) tanks were compared with reference to ATP, carbon content, and physical properties. Mean values for bacterial numbers and ATP were about twice as high in the MES, carbon content was slightly higher in the artificial systems, and the sediment of the MES was enriched for larger grain sizes and depleted of silt and clay. In the natural marsh sediment, ATP was strongly related (Pearson's correlation coefficient) to granulometric (or physical) factors, whereas ATP was related to biological factors in the MES. Approximately 61% of the variation in ATP concentrations in the marsh sediments could be accounted for by the parameters measured. Only half that amount could be explained on the basis of those same variables in the MES. Furthermore, one of the four tanks was significantly different from the other three in terms of ATP concentration, and an anomaly in respect to silt-clay content was demonstrated in another. Careful control and some type of equilibrating procedure will be necessary before the MES can be employed as replicate units. PMID- 16345329 TI - Effects of no. 2 fuel oil, nigerian crude oil, and used crankcase oil on attached algal communities: acute and chronic toxicity of water-soluble constituents. AB - Water extracts of a no. 2 fuel oil, a Nigerian crude oil, and used crankcase oil were examined for their effects on algal communities in experiments lasting several weeks conducted under near-natural conditions. No. 2 fuel oil extracts depressed algal biomass (chlorophyll a) and resulted in blue-green algal (cyanobacterial) dominance and decreased diatom occurrence. Changes in concentrations of chlorophyll c, which was specific for diatoms in this work, and phycocyanin, which was specific for blue-green algae, confirmed the observations. Used crankcase oil extracts also depressed biomass, but Nigerian crude extracts did not, and both these extracts had less effect on community composition than did no. 2 fuel oil extracts. Photosynthetic C incorporation was both stimulated and depressed by exposure to extracts with hydrocarbon concentrations 0.038 to 0.124 mg/liter. Short-term exposure to higher concentrations (1.17 to 15.30 mg of hydrocarbons per liter) of no. 2 fuel oil extracts depressed photosynthetic C incorporation by Vaucheria-dominated communities in all tests but one. Toxicity was greater from extracts prepared in the light than from extracts prepared in the dark. PMID- 16345330 TI - Enumeration, isolation, and characterization of beggiatoa from freshwater sediments. AB - An accurate most-probable-number enumeration method was developed for counting the number of Beggiatoa trichomes from various freshwater sediments. The medium consisted of extracted hay, diluted soil extract, 0.05% acetate, and 15 to 35 U of catalase per ml. The same enrichment medium, but without the acetate, was the best enrichment medium from which to obtain pure cultures because it supported good growth of the beggiatoas without allowing them to be overgrown by other bacteria. A total of 32 strains of Beggiatoa were isolated from seven different freshwater habitats and partially characterized. The strains were separated into five groups based on several preliminary characteristics. Four of the groups contained cells with trichomes of approximately the same diameter (1.5 to 2.7 mum) and may be Beggiatoa leptomitiformis or an unnamed species. The fifth group appeared to be Beggiatoa alba. With the exception of three strains, all of the strains deposited sulfur in the presence of hydrogen sulfide, and all strains grew heterotrophically and deposited poly-beta-hydroxybutyrate and volutin when grown on acetate supplemented with low concentrations of other organic nutrients. Thin sections of sulfur-bearing trichomes indicated that the sulfur granules were external to the cytoplasmic membrane and that they were surrounded by an additional membrane. PMID- 16345332 TI - Improved methods for detecting enteric viruses in oysters. AB - [This corrects the article on p. 127 in vol. 36.]. PMID- 16345331 TI - Diversity of Rhizobium leguminosarum in the Palouse of Eastern Washington. AB - Serology, antibiotic resistance, litmus milk reaction, and ability to fix nitrogen in peas were methods employed in studying the diversity of Rhizobium leguminosarum. The two serogroups identified were related to the position on the soil slope where isolated were taken. PMID- 16345333 TI - Minimal Water Activity for Enterotoxin A Production and Growth of Staphylococcus aureus. AB - [This corrects the article on p. 378 in vol. 36.]. PMID- 16345334 TI - Electrode system for the determination of microbial populations. AB - Determinations of microbial populations were carried out by using a new electrode system composed of two electrodes. Each electrode was constructed from a platinum anode and a silver peroxide cathode. The anode of the reference electrode was covered with cellulose dialysis membrane. The response time of the electrode system was 15 min in culture broth, and current differences between the two electrodes were proportional to populations of microbial cells in cultures of Saccharomyces cerevisiae and Lactobacillus fermentum. Current differences were reproducible; the average relative error was 5%. Furthermore, cell populations of S. cerevisiae in a fermentor could be continuously estimated by using this electrochemical method. PMID- 16345336 TI - Estimation of sediment denitrification rates at in situ nitrate concentrations. AB - The denitrification rates in a marine sediment, estimated by using N-nitrate, V(max), K(m), and sediment nitrate concentrations, were 12.5 and 2.0 nmol of N(2) N cm day at 0 to 1 and 1 to 3 cm, respectively, at 12 degrees C. The total rate was 165 nmol of N(2)-N m day. PMID- 16345335 TI - Germ Tube Growth Inhibitor from Cronartium comandrae Aeciospores. AB - Two compounds showing self-inhibitory action during germination of aeciospores of the comandra blister rust fungus (Cronartium comandrae Pk.) were extracted from these aeciospores by shaking with 0.2 M NH(4)HCO(3) (pH 7.8) for 4 h. One of these, the germination self inhibitor (D. A. Eppstein and F. H. Tainter, Phytopathology 66:1395-1397, 1976), was removed from the ammonium bicarbonate buffer by using chloroform. The water layer which remained contained a substance which, at ca. 10 M concentration, had no apparent effect on germ tube emergence but which inhibited normal germ tube growth. Linear germ tube growth ceased or a dendritic or vesicular pattern of growth resulted, depending on the concentration of inhibitor added to extracted germinating spores. The germ tube growth inhibitor appears to be a peptide with a molecular weight of ca. 2,000. PMID- 16345338 TI - Influence of substratum characteristics on the attachment of a marine pseudomonad to solid surfaces. AB - The attachment of a marine Pseudomonas sp. to a variety of surfaces was investigated, and the number of bacteria which became attached was related to the surface charge and degree of hydrophobicity of the substratum. Large numbers of bacteria attached to hydrophobic plastics with little or no surface charge [Teflon, polyethylene, polystyrene, poly(ethylene terephthalate)]; moderate numbers attached to hydrophilic metals with a positive (platinum) or neutral (germanium) surface charge; and very few attached to hydrophilic, negatively charged substrata (glass, mica, oxidized plastics). The results suggest that both electrostatic and hydrophobic interactions are involved in bacterial attachment. PMID- 16345337 TI - Growth of a bacterium under a high-pressure oxy-helium atmosphere. AB - Growth of a barotolerant marine organism, EP-4, in a glutamate medium equilibrated with an oxy-helium atmosphere at 500 atmospheres (atm; total pressure) (20 degrees C) was compared with control cultures incubated at hydrostatic pressures of 1 and 500 atm. Relative to the 1-atm control culture, incubation of EP-4 at 500 atm in the absence of an atmosphere resulted in an approximately fivefold reduction in the growth rate and a significant but time variant reduction in the rate constants for the incorporation of substrate into cell material and respiration. Distinct from the pressurized control and separate from potential effects of dissolution of helium upon decompression of subsamples, exposure of the organism to high-pressure oxy-helium resulted in either a loss of viability of a large fraction of the cells or the arrest of growth for one-third of the experimental period. After these initial effects, however, the culture grew exponentially at a rate which was three times greater than the 500-atm control culture. The rate constant for the incorporation of substrate into cell material was also enhanced twofold in the presence of high-pressure oxy-helium. Dissolved oxygen was well controlled in all of the cultures, minimizing any potential toxic effects of this gas. PMID- 16345339 TI - Effect of Ammonium Chloride and Methionine Sulfoximine on the Acetylene Reduction of Detached Root Nodules of Peas (Pisum sativum). AB - Acetylene-reducing activity of detached pea nodules was determined by submerging the nodules in buffer solution [tris(hydroxymethyl)aminomethane-hydrochloride, pH 7.4] containing 100 mM sodium succinate and incubating under a gas phase of 90% O(2) and 10% C(2)H(2). The nitrogenase activity was 4 to 8 mumol of C(2)H(4) formed per g of nodule fresh weight per h and remained constant for at least 4 h. Addition of NH(4)Cl to the buffer solution (at a concentration of 10 mM or more) resulted in a significant decrease of nitrogenase activity, which was more pronounced at higher concentrations of ammonium chloride. The inhibition of nitrogenase activity by NH(4)Cl was reversible; when the NH(4)Cl-containing buffer solution was replaced by buffer without NH(4)Cl, the original activity was partly restored. Treatment of the nodules with NH(4)Cl had almost no effect on the amount of nitrogenase, as measured by the acetylene-reducing activity of ethyl-enediaminetetraacetate-toluene-treated bacteroid suspensions. The effect of NH(4)Cl was largely eliminated by simultaneous addition of 10 mM methionine sulfoximine to the assay solution. This suggests that the assimilation of ammonium ions by glutamine synthetase controls the functioning of nitrogenase activity in the nodules. However, no effect of glutamine, glutamate, or aspartate on the acetylene reduction by detached nodules could be detected. PMID- 16345340 TI - Types and distribution of anaerobic bacteria in the large intestine of pigs. AB - An examination was made of various sites along the length of the swine large intestine, using strictly anaerobic culture methods. Sites were separated by differential washing into fractions described as lumenal content, lumenal surface layer, and intestinal wall tissue. Direct microscopic clump counts averaged 13.3 x 10 organisms per g (dry weight) of material in the lumenal content, 14.0 x 10 in the surface layer, and 5.1 x 10 in the intestinal wall tissue. Both direct microscopic counts and viable culture counts were higher from the lumenal content and surface layer than from the intestinal tissue at all sites sampled in the intestine. Cultural counts averaged 56.2% of the direct microscopic counts in lumenal content and surface layer and 20.2% in intestinal tissue. Over 90% of the bacteria isolated were gram positive and consisted mainly of gram-positive cocci, lactobacilli, eubacteria, and clostridia. Of 192 isolates recovered, only 124 could be assigned to recognized species. PMID- 16345341 TI - Increased susceptibility of rainbow trout to infectious hematopoietic necrosis virus after exposure to copper. AB - Exposure of rainbow trout to sublethal levels of copper in water increased their susceptibility to infectious hematopoietic necrosis virus. In most instances, the percent mortality was twice as great in the stressed groups compared with those groups which were not stressed but received the same virus dose. Although the level of copper in the water influenced the mortality rates, the length of exposure did not prove to be critical, as similar results were obtained after 1, 3, 5, 7, or 9 days of exposure. When different virus challenges were employed, the percent mortalities were again greater in the stressed fish at all virus doses tested, and at one dose level mortalities were noted in the stressed group but not in the untreated group. PMID- 16345342 TI - Extraction and quantification of solutes in solidified agar culture media. AB - A method is described for determining the concentration of certain solutes in solidified culture media. The method is based upon the finding that under specified conditions the concentration of solute in an agar gel (C(g)) is related to the concentration of solute in a centrifugally extracted gel supernatant (C(s)) by the ratio, C(g)/C(s), which is characteristic for each solute. The method avoids direct assays of the gels and instead involves assaying the supernatants from inoculated and uninoculated (control) gels with conventional liquid assay techniques and then calculating solute concentrations in the inoculated gels by use of the C(g)/C(s) ratios determined from the controls. Uninoculated agar gels containing known concentrations of various solutes and similar gels inoculated with Neurospora crassa or Escherichia coli were centrifuged at various times, and the supernatants were assayed for solute concentrations. The solute concentrations in the supernatants from the inoculated gels multiplied by the C(g)/C(s) ratios for those solutes determined at the same times for the uninoculated controls gave calculated solute concentrations in the inoculated gels. The differences between these calculated solute concentrations and those initially present in the inoculated gels indicated the amounts of solutes utilized from the gels by the microorganisms at various incubation times. PMID- 16345343 TI - Determination of the carbon-bound electron composition of microbial cells and metabolites by dichromate oxidation. AB - The applicability of the silver sulfate-acid dichromate oxidation (chemical oxygen demand) method for determining the carbon-bound electron compositions of microbial cells, substrates, and metabolic by-products was evaluated. An approach for approximating the carbon-bound electron composition of microbial cells from CHN data is also presented. Ten aliphatic and aromatic carboxylic acids, 17 amino acids, and 8 sugars generally gave 96 to 101% (mainly >/=98%) recovery with 0.0625 N dichromate (digestion mixture of 10 ml of sample-10 ml of 0.25 N dichromate-20 ml of Ag(2)SO(4)-amended concentrated H(2)SO(4)). Recoveries of nicotinic acid (5%) and methionine (65%) were incomplete; arginine (125%) and two purine and three pyrimidine bases (105 to 120%) were overestimated. The validity of 0.0625 N dichromate for determining the carbon-bound electron composition of bacterial cells was supported by theoretical analysis of the carbon-bound electron composition of hypothetical bacterial cell material (defined monomer composition) and by the compatibility of elemental and dichromate oxidation derived carbon-bound electron compositions of typical bacterial cells. PMID- 16345344 TI - Monensin and dichloroacetamide influences on methane and volatile Fatty Acid production by rumen bacteria in vitro. AB - The effect of monensin (0 or 33 mug/g of diet) upon rumen fermentation in the presence and absence of methanogenesis was determined in vitro by using mixed rumen organisms continuously cultured for 17 days. Methane was inhibited by dichloroacetamide (DCA; 32 mg/day) or by a pH of 5.1. Monensin effected a significant decrease in the ratio of acetic to propionic acid in the presence or absence of methanogenesis. In the absence of methanogenesis, the decrease in the ratio of acetic to propionic acid was entirely the result of increased propionic acid, whereas in the presence of methanogenesis the decrease in the ratio was the result of a combination of decreased acetic acid and increased propionic acid. There was a complementary interaction between monensin and DCA on volatile fatty acid production (expressed as millimoles of carbon per day). Addition of monensin to DCA-treated cultures resulted in the production of more acid; however, monensin and DCA had no beneficial effect on total carbon formed as acid and gases as compared with nonsupplemented control cultures. The monensin and DCA also resulted in greater digestion of neutral detergent fiber and less accumulation of formic acid and hydrogen as end products than did DCA alone. l Lactic acid was produced in small but significantly greater amounts by the low-pH cultures, which also had less volatile fatty acid carbon formed from the fiber fraction of the forage supplied. PMID- 16345345 TI - Immobilization of Glucose Isomerase-Containing Streptomyces phaeochromogenes Cells in Fine-Particle Form. AB - A new preparation method for immobilizing Streptomyces phaeochromogenes cells in fine-particle form was investigated using radiation-induced polymerization at low temperatures with previously salted out hydrophilic monomers. Using this method, it was found that the glucose isomerase activity of the immobilized cell particles was markedly higher than that of immobilized cells in block form obtained without salting out of the monomer. The diameter of the particles was varied by changing the irradiation temperature or the concentrations of monomer and salt. The magnitude of the enzymatic activity increased with decreasing particle diameter. K(m) values of the immobilized cell particles were close to that of the intact cell. These facts suggested that the cells were trapped on the surface of the particle. PMID- 16345346 TI - Characterization of Plasmid Deoxyribonucleic Acid in Streptococcus lactis subsp. diacetylactis: Evidence for Plasmid-Linked Citrate Utilization. AB - The use of Streptococcus diacetylactis as a flavor producer in dairy fermentations is dependent upon its ability to produce diacetyl from citrate. Treatment of S. diacetylactis strains 18-16 and DRC1 with acridine orange resulted in the conversion of approximately 2% of the DRC1 population and 20% of the 18-16 population to citrate negative, which is indicative of the involvement of plasmid deoxyribonucleic acid (DNA). Growth in the presence of acridine orange also resulted in the appearance of 2% lactose-negative derivatives in S. diacetylactis 18-16 and 99% lactose-defective, proteinase-negative derivatives in S. diacetylactis DRC1. Cesium chloride-ethidium bromide equilibrium density gradients of cleared lysate material from each strain revealed the presence of covalently closed circular DNA. Samples of this covalently closed circular DNA were subjected to agarose gel electrophoresis to determine the plasmid composition of each strain. S. diacetylactis 18-16 was found to possess six plasmids, of approximately 41, 28, 6.4, 5.5, 3.4, and 3.0 megadaltons (Mdal). S. diacetylactis DRC1 contained six plasmids, of approximately 41, 31, 18, 5.5, 4.5, and 3.7 Mdal. Variants of S. diacetylactis 18-16 which failed to produce acetoin plus diacetyl from citrate (citrate negative) were missing a 5.5-Mdal plasmid. Lactose-negative mutants of the same strain were devoid of a 41-Mdal plasmid. Lactose-defective, proteinase-negative mutants of S. diacetylactis DRC1 were missing a 31-Mdal plasmid. The citrate-negative mutants of S. diacetylactis DRC1 isolated in this study did not possess a 5.5-Mdal plasmid. Thus, we have evidence that there is a correlation between the ability to utilize citrate and the presence of a 5.5-Mdal plasmid. A relationship was also noted between lactose fermentation and proteinase activity and plasmid DNA in S. diacetylactis. PMID- 16345347 TI - Production of Glucose Isomerase by Streptomyces flavogriseus. AB - A microorganism that produces glucose isomerase was isolated from soil and identified as a strain of Streptomyces flavogriseus. The organism produced a large quantity of glucose isomerase when grown on straw hemicellulose, xylan, xylose, and H(2)SO(4) hydrolysate of ryegrass straw. The organism produced glucose isomerase both intra- and extra-cellularly. The highest level of intracellular glucose isomerase (3.5 U/ml) was obtained in about 36 h by a culture grown on straw hemicellulose; the extracellular enzyme (1.5 U/ml) appeared in cultures grown for about 72 h. About equal levels of enzyme were produced in cultures grown on straw hemicellulose, xylan, xylose, and H(2)SO(4) hydrolysate of straw, but production of the enzyme was drastically reduced when the organism was grown on other carbon sources. As a nitrogen source, corn steep liquor produced the best results. Soy flour extract, yeast extract, and various peptones also were adequate substrates for glucose isomerase production. Addition of Mg, Mn, or Fe to the growth medium significantly enhanced enzyme production. The organism, however, did not require Co, which is commonly required by microorganisms used in the production of glucose isomerase. PMID- 16345348 TI - Mode of attack on orchardgrass leaf blades by rumen protozoa. AB - Leaf blade sections of orchardgrass were incubated with rumen fluid and examined by scanning and transmission electron microscopy for the mode of attack on tissues by rumen protozoa. Rumen protozoa resembling Epidinium ecaudatum from caudatum degraded forage tissue in diluted, whole rumen fluid suspensions of microbes containing 1.6 mg of streptomycin per ml, which inhibited bacterial fiber-digesting activity. Cell walls of mesophyll, parenchyma bundle sheath, and epidermis became swollen and frayed to reveal a microfibrillar network and loss of electron density, indicating partial degradation. Then the protozoa ingested whole cells and fragments of cell walls with the aid of their cilia. Plant cells with partially degraded walls as well as chloroplasts without walls were present within the protozoa. These entodiniomorphs digested orchardgrass leaves by partially degrading the plant cell walls apparently by extracellular enzymes and then ingestion of the plant cells and cell wall fragments. PMID- 16345349 TI - Association of actinomycete-like bacteria with soil-feeding termites (termitidae, termitinae). AB - Electron microscopy of the hindgut showed that actinomycete-like bacteria were the principal microbial associates of two African species of soil-feeding termites. Elongated cuticular spines provided attachment. PMID- 16345350 TI - Symbiotic Cellulose Degradation in Green Turtles, Chelonia mydas L. AB - A postgastric, fermentative breakdown of structural plant tissue was demonstrated for green turtles. About 90% cellulose was hydrolyzed. Bacterial and protozoan numbers compared with those of the rumen. PMID- 16345351 TI - Nitrogen fixation associated with the rice plant grown in water culture. AB - Acetylene reduction activity of intact rice plants was measured in closed assay chambers with plants grown in water culture. Acetylene was added to the liquid medium, and the ethylene formed was measured from both gas and liquid phases. After cutoff of mineral nitrogen supply and inoculation of fresh soil, rice plants grown from the seedling stage in water culture exhibited acetylene reduction activity after a lag period. However, rice plants grown in a paddy field and transferred to water culture were more suitable for N(2) fixation studies because of their higher, less variable acetylene reduction activity. The time course of acetylene reduction was monitored by continuous circulation of gas between the gas phase and the liquid phase, and the result showed an initial 2- or 3-h period of lower activity, followed by increased and almost constant activity up to 24 h. The effects on acetylene reduction activity of aeration, ammonium, chloramphenicol, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea addition are reported. Ammonium was inhibitive at 0.33 mM, and its depressive effect was alleviated by ammonium uptake by the plants. PMID- 16345352 TI - Grouping of lactic streptococci by gel electrophoresis of soluble cell extracts. AB - Soluble cell proteins obtained from 35 strains of lactic streptococci were examined by gel electrophoresis. A mathematical analysis of the densitometer scans of the gels enabled strains to be grouped according to their overall similarity. Strains which were known to be variants of the same parent strain fell into the same group, supporting the validity of the method. It is suggested that strains which are alike according to their gel electrophoretic patterns when grown under standard conditions have an overall phenotypic similarity and that this indicates a similarity in genotype. The relevance of this to selection of strains of lactic streptococci for cheesemaking is discussed. PMID- 16345353 TI - Nitrogen fixation by thermophilic blue-green algae (cyanobacteria): temperature characteristics and potential use in biophotolysis. AB - Thermophilic, nitrogen-fixing, blue-green algae (cyanobacteria) were investigated for use in biophotolysis. Three strains of Mastigocladus laminosus were tested and were found to be equally effective in biophotolysis as judged by nitrogenase activity. The alga, M. laminosus NZ-86-m, which was chosen for further study, grew well in the temperature range from 35 to 50 degrees C, with optimum growth at 45 degrees C, at which temperature acetylene reduction activity was also greatest. The maximum tolerable temperature was 55 degrees C. Acetylene reduction activity was saturated at a light intensity of 1 x 10 ergs cm s. Atmospheric oxygen tension was found to be slightly inhibitory to acetylene reduction of both slowly growing and exponentially growing cultures. Nonsterile continuous cultures, which were conducted to test problems of culture maintenance, could be operated for 2 months without any significant decrease in nitrogenase activity or contamination by other algae. Nitrogen-starved cultures of M. laminosus NZ-86-m produced hydrogen at comparable rates to Anabaena cylindrica. The conversion efficiency of light to hydrogen energy at maximum rates of hydrogen production was 2.7%. PMID- 16345354 TI - Poly-beta-Hydroxybutyrate Accumulation as a Measure of Unbalanced Growth of the Estuarine Detrital Microbiota. AB - The procaryotic endogenous storage material poly-beta-hydroxybutyrate (PHB) can be induced to accumulate in the estuarine detrital microbiota under conditions which suggest unbalanced growth, such as limitation of a critical factor(s) in the presence of carbon and energy sources. Changes in PHB-to-lipid phosphate ratios detected in field samples can be mimicked in the laboratory with common estuarine stresses. Acute anoxia or low pH induces conditions of no growth with depression of both the synthesis and catabolism of PHB without change in the lipid phosphate. Balanced growth induced by nutrients increases the lipid phosphate, depresses PHB synthesis, and stimulates PHB catabolism, resulting in a low ratio of PHB to lipid phosphate. Unbalanced growth induced to a small extent by high salinity or much more readily by dark upland runoff water results in rapid accumulation of PHB and slowing of PHB catabolism with little change in lipid phospate. Unbalanced growth conditions result in high PHB-to-lipid phosphate ratios in the detrital microbiota. PMID- 16345355 TI - Microbial biomass and activity distribution in an anoxic, hypersaline basin. AB - The Orca Basin is a hypersaline depression in the northern Gulf of Mexico with anoxic conditions observed in the lower 200 m of the water column. Measurements of adenosine 5'-triphosphate, heterotrophic potential, and uridine uptake made above and across the interface into the anoxic zone revealed the presence of an active microbial population approximately 100 m above the interface. Biomass and activity decreased at and just below the interface but increased near the bottom, consistent with similar observations made in the Cariaco Trench. The maximum adenosine 5'-triphosphate concentration above the interface of 5.9 ng/liter (2,173 m) is about eight times greater than the value found in oxygenated waters of corresponding depth in the absence of an anoxic zone. The maximum adenosine 5' triphosphate concentration in the anoxic zone is approximately 15 times greater than that found in oxygenated water of similar depth, suggesting anoxia will support the development of a larger bacterial population. Our findings suggest that autotrophic bacteria may be the dominant physiological group in the region just above the interface. PMID- 16345356 TI - Dialysis continuous process for ammonium-lactate fermentation: improved mathematical model and use of deproteinized whey. AB - Separate terms for substrate limitation and product inhibition were incorporated into an equation describing the rate of cell growth for the steady-state fermentation of lactose to lactic acid with neutralization to a constant pH by ammonia. The equation was incorporated into a generalized mathematical model of a dialysis continuous process for the fermentation, developed previously, in which the substrate is fed into the fermentor and the fermentor contents are dialyzed through a membrane against water. The improved model was used to simulate the fermentation on a digital computer, and the results agreed with previous experimental tests using whole whey as the substrate. Further simulations were then made to guide experimental tests using deproteinized whey as the substrate. Dried cheese-whey ultrafiltrate was rehydrated with tap water to contain 242 mg of lactose per ml, supplemented with 8 mg of yeast extract per ml, charged into a 5-liter fermentor without sterilization, adjusted in pH (5.5) and temperature (44 degrees C), and inoculated with an adapted culture of Lactobacillus bulgaricus. The fermentor and dialysate circuits were connected, and a series of steady-state conditions was managed nonaseptically for 71 days. The fermentation of deproteinized whey relative to whole whey, with both highly concentrated, resulted in similar extents of product accumulation but at a lesser rate. PMID- 16345357 TI - Apparent Involvement of a Plasmid in Phaseotoxin Production by Pseudomonas phaseolicola. AB - Three naturally occurring toxigenic strains (HB-36, G-50, and HB-33), one nontoxigenic strain (HB-20), and one ultraviolet light-induced toxinless mutant (G-50 Tox) of Pseudomonas phaseolicola were examined by dye-buoyant density equilibrium centrifugation for the presence of plasmid deoxyribonucleic acid. All strains contained plasmid deoxyribonucleic acid. Comparison of the plasmid deoxyribonucleic acid of different strains by agarose gel electrophoresis showed that strain G-50 harbored three plasmids, whereas the rest of the strains contained two plasmids each. Irrespective of their toxigenicity, all strains shared the large-sized first plasmid band, but differed with respect to other plasmids. Restriction endonuclease analyses of the plasmids indicated that a 22.50-megadalton plasmid was common to two of the toxigenic strains (HB-36 and G 50). However, strain HB-33, which is also toxigenic, contained a much smaller plasmid (4.23 megadaltons). It is hypothesized that this small plasmid may have arisen by a recombination event from a larger plasmid. PMID- 16345358 TI - Comparison of substrate affinities among several rumen bacteria: a possible determinant of rumen bacterial competition. AB - Five rumen bacteria, Selenomonas ruminantium, Bacteroides ruminicola, Megasphaera elsdenii, Streptococcus bovis, and Butyrivibrio fibrisolvens were grown in continuous culture. Estimates of substrate affinities were derived from Lineweaver-Burk plots of dilution rate versus substrate concentration. Each bacterium was grown on at least four of the six substrates: glucose, maltose, sucrose, cellobiose, xylose, and lactate. Wide variations in substrate affinities were seen among the substrates utilized by a species and among species for the same substrate. These wide differences indicate that substrate affinity may be a significant determinant of bacterial competition in the rumen where soluble substrate concentrations are often low. Growth of these bacteria in continuous culture did not always follow typical Michaelis-Menten kinetics. Inflated theoretical maximum growth rates and non-linear Lineweaver-Burk plots were sometimes seen. Maintenance energy expenditures and limitation of growth rate by factors other than substrate concentration (i.e., protein synthesis) are discussed as possible determinants of these deviations. PMID- 16345359 TI - Comparison of maintenance energy expenditures and growth yields among several rumen bacteria grown on continuous culture. AB - Maintenance energy expenditures were mesured for five rumen bacteria, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Bacteroides ruminicola, Megasphaera elsdenii, and Streptococcus bovis, by using a complex medium with glucose as the carbon source. Large differences (as high as 8.5-fold) in maintenance energy expenditures were seen among these bacteria. The suggestion is made that maintenance requirements could be a significant determinant of bacterial competition in the rumen. Theoretical maximum growth yields, calculated from double reciprocal plots of yield versus dilution rate, were compared to theoretical Y(ATP) values in order to estimate minimum molar adenosine 5' triphosphate yields from glucose for each bacterium. Results showed that relative yield among the bacteria was growth rate dependent. At high dilution rates, both S. ruminantium and S. bovis produced lactate as their principal fermentation product. At lower dilution rates very little lactate was formed and growth yields increased. Acetate and ethanol were the predominant fermentation products of S. bovis at low dilution rates. Other workers have shown that S. ruminantium produces acetate and propionate at low growth rates. PMID- 16345360 TI - Effects of combinations of substrates on maximum growth rates of several rumen bacteria. AB - Five rumen bacteria, Selenomonas ruminantium, Bacteroides ruminicola, Megasphaera elsdenii, Butyrivibrio fibrisolvens, and Streptococcus bovis were grown in media containing nonlimiting concentrations of glucose, sucrose, maltose, cellobiose, xylose and/or lactate. Each bacterium was grown with every substrate that it could ferment in every possible two-way combination. Only once did a combination of substrates result in a higher maximum growth rate than that observed with either substrate alone. Such stimulations of growth rate would be expected if specific factors unique to individual substrates (transport proteins and/or enzymes) were limiting. Since such synergisms were rare, it was concluded that more general factors limit maximum growth rates in these five bacteria. PMID- 16345361 TI - Hydrogen Evolution from Alfalfa and Clover Nodules and Hydrogen Uptake by Free Living Rhizobium meliloti. AB - A series of Rhizobium meliloti and Rhizobium trifolii strains were used as inocula for alfalfa and clover, respectively, grown under bacteriologically controlled conditions. Replicate samples of nodules formed by each strain were assayed for rates of H(2) evolution in air, rates of H(2) evolution under Ar and O(2), and rates of C(2)H(2) reduction. Nodules formed by all strains of R. meliloti and R. trifolii on their respective hosts lost at least 17% of the electron flow through nitrogenase as evolved H(2). The mean loss from alfalfa nodules formed by 19 R. meliloti strains was 25%, and the mean loss from clover nodules formed by seven R. trifolii strains was 35%. R. meliloti and R. trifolii strains also were cultured under conditions that were previously established for derepression of hydrogenase synthesis. Only strains 102F65 and 102F51 of R. meliloti showed measurable activity under free-living conditions. Bacteroids from nodules formed by the two strains showing hydrogenase activity under free-living conditions also oxidized H(2) at low rates. The specific activity of hydrogenase in bacteroids formed by either strain 102F65 or strain 102F51 of R. meliloti was less than 0.1% of the specific activity of the hydrogenase system in bacteroids formed by H(2) uptake-positive Rhizobium japonicum USDA 110, which has been investigated previously. R. meliloti and R. trifolii strains tested possessed insufficient hydrogenase to recycle a substantial proportion of the H(2) evolved from the nitrogenase reaction in nodules of their hosts. Additional research is needed, therefore, to develop strains of R. meliloti and R. trifolii that possess an adequate H(2)-recycling system. PMID- 16345362 TI - Colorimetric enzyme-linked immunosorbent assay for the identification of strains of Rhizobium in culture and in the nodules of lentils. AB - An indirect enzyme-linked immunosorbent assay has been developed to identify strains of Rhizobium in culture and in lentil nodules. The test can be used on cells from both fresh and frozen nodules obtained from plants grown either in a growth chamber or in the field. Test results were confirmed by immunofluorescence. The enzyme-linked immunosorbent assay technique can be used for field studies and requires less antisera than other serological techniques. PMID- 16345363 TI - Niche in Pasture-Fed Ruminants for the Large Rumen Bacteria Oscillospira, Lampropedia, and Quin's and Eadie's Ovals. AB - The little-studied large bacteria Oscillospira, Lampropedia, and ovals attach rapidly in large numbers to the cuticular surface of clover and grass leaves in the rumen. The cuticle of green leaves may constitute a specific niche for these bacteria. PMID- 16345365 TI - Production, Characterization, and Partial Amino Acid Sequence of Xylanase A from Schizophyllum commune. AB - [This corrects the article on p. 804 in vol. 36.]. PMID- 16345364 TI - Passage and Survival of Chlamydospores of Phytophthora cinnamomi Rands, the Causal Agent of Forest Dieback Disease, Through the Gastrointestinal Tracts of Termites and Wild Birds. AB - Chlamydospores of Phytophthora cinnamomi Rands have been shown to survive in the intestinal tracts of termites (Nasutitermes exitiosus) and two species of forest birds indigenous to West Australian jarrah forests. Viable chlamydospores were recovered from bird feces within the normal rate of passage time for food through the gut. The above factors would allow these creatures to function as vectors for the spores. PMID- 16345366 TI - Conversion of biovolume measurements of soil organisms, grown under various moisture tensions, to biomass and their nutrient content. AB - Direct microscopic measurements of biomass in soil require conversion factors for calculation of the mass of microorganisms from the measured volumes. These factors were determined for two bacteria, five fungi, and a yeast isolated from soil. Moisture stress conditions occurring in nature were simulated by growth in two media using shake cultures, on agar plates, and on membranes held at 34, 330, and 1,390 kPa of suction. The observed conversion factors, i.e., the ratio between dry weight and wet volume, generally increased with increasing moisture stress. The ratios for fungi ranged from 0.11 to 0.41 g/cm with an average of 0.33 g/cm. Correction of earlier data assuming 80% water and a wet-weight specific gravity of 1.1 would require a conversion factor of 1.44. The dry-weight specific gravity of bacteria and yeasts ranged from 0.38 to 1.4 g/cm with an average of 0.8 g/cm. These high values can only occur at 10% ash if no more than 50% of the cell is water, and a specific conversion factor to correct past data for bacterial biomass has not yet been suggested. The high conversion factors for bacteria and yeast could not be explained by shrinkage of cells due to heat fixing, but shrinkage during preparation could not be completely discounted. Moisture stress affected the C, N, and P content of the various organisms, with the ash contents increasing with increasing moisture stress. Although further work is necessary to obtain accurate conversion factors between biovolume and biomass, especially for bacteria, this study clearly indicates that existing data on the specific gravity and the water and nutrient content of microorganisms grown in shake cultures cannot be applied when quantifying the soil microbial biomass. PMID- 16345367 TI - Extracellular Proteases of Mucor pusillus. AB - Mucor pusillus was grown in different media for a period of 92 h, and the media were investigated for both milk-clotting and protease activities. It was observed that the ratio of extracellular milk-clotting activity to protease activity was the highest for 3% corn steep liquor containing 1% glucose as the source of carbon. Variation of both milk-clotting and protease activities was studied during the growth of the organism in the medium stated above. Separation of protease was carried out by ion-exchange chromatography at pH 8.0. Fractions collected were assayed for both activities simultaneously. The findings suggested that, instead of only one major acid protease, as reported by previous workers, two major acid proteases were produced. One of them had significant rennin-like activity, and the other lacked it. The former could be assumed to be the enzyme reported and studied by previous workers. The existence of two proteases was further confirmed by the appearance of two protease activity bands on polyacrylamide gels after electrophoresis. An attempt was made to separate the rennin-like enzyme from nonspecific protease activity by ammonium sulfate fractionation followed by ion-exchange chromatography at pH 6.0. The results indicated that the nonspecific protease activity due to the enzyme that lacked rennin action was substantially removed by the ammonium sulfate fractionation. PMID- 16345368 TI - Bacteria associated with the surface and gut of marine copepods. AB - Little is known about the nature of bacteria associated with the surface and gut of marine copepods, either in laboratory-reared animals or in the natural environment. Nor is it known whether such animals possess a gut flora. The present report deals with studies of microorganisms isolated from healthy, laboratory-reared copepods of the species Acartia tonsa Dana, from several species of wild copepods collected from a marine or estuarine environment, and from laboratory dishes containing moribund copepods. Evidence for a unique gut flora in laboratory-reared animals is presented; the predominant bacteria were represented by the genus Vibrio. Other organisms such as Pseudomonas and Cytophaga were found less abundantly associated with the copepods and not specifically associated with the gut. PMID- 16345369 TI - Method for Measuring Rates of NH(4) Turnover in Anoxic Marine Sediments, Using a N-NH(4) Dilution Technique. AB - A method is described for the determination of the net and total rates of NH(4) production and NH(4) incorporation at different depths in an anoxic marine sediment. N-NH(4) was added to the sediment NH(4) pool, and the N content was assayed after 0, 2, and 5 days of incubation. The pool size changed during incubation; this change in pool size is incorporated into a model which predicts the dynamics of N-NH(4) dilution. A simple microdiffusion of NH(3) was followed by an emission spectrometry analysis of N content. This procedure avoided all problems of cross-contamination. The model was tested and rates were measured in four sediment cores, at seven different depths. The high correlation coefficients (mean, 0.96 for the 0- to 2-, 2- to 4-, 4- to 6-, and 6- to 8-cm sediment fractions) indicated that the model was correct and that the measured rates were valid. The immediate distribution of N-NH(4) between interstitial and exchangeable NH(4) pools indicated that it was the combined pool that was turning over. In the 0- to 2-cm fraction at 17 degrees C the net rate of NH(4) production was 274 (standard deviation, 31) nmol cm day, and the mean total rate of NH(4) production was 309 (standard deviation, 39) nmol cm day; both rates decreased to <1% of these values in the 12- to 14-cm fractions. PMID- 16345370 TI - Polyacrylamide-entrapped Rhizobium as an inoculant for legumes. AB - Pot experiments showed that Rhizobium japonicum cells entrapped in a polyacrylamide gel could be used as an inoculant for soybeans and compared favorably to laboratory-made peat base inoculant containing the same bacterial strain. PMID- 16345371 TI - High numbers of prosthecate bacteria in pulp mill waste aeration lagoons. AB - Prosthecate bacteria comprised 0.6 to 10.5% of the bacterial community in samples from 11 pulp mill waste aeration lagoons. Because of their distinct morphology, the genera Ancalomicrobium, Caulobacter, Prosthecobacter, Prosthecomicrobium, Stella, and Hyphomicrobium or Hyphomonas could be identified and enumerated by direct microscopic examination. Monthly samples from one lagoon showed that several genera varied from undetectable to predominant among the appendaged organisms. Temperature (season), type of wood pulped, and pulping process did not significantly affect the density of prosthecate bacteria. PMID- 16345372 TI - Plant Growth Substances Produced by Azospirillum brasilense and Their Effect on the Growth of Pearl Millet (Pennisetum americanum L.). AB - Azospirillum brasilense, a nitrogen-fixing bacterium found in the rhizosphere of various grass species, was investigated to establish the effect on plant growth of growth substances produced by the bacteria. Thin-layer chromatography, high pressure liquid chromatography, and bioassay were used to separate and identify plant growth substances produced by the bacteria in liquid culture. Indole acetic acid and indole lactic acid were produced by A. brasilense from tryptophan. Indole acetic acid production increased with increasing tryptophan concentration from 1 to 100 mug/ml. Indole acetic acid concentration also increased with the age of the culture until bacteria reached the stationary phase. Shaking favored the production of indole acetic acid, especially in a medium containing nitrogen. A small but biologically significant amount of gibberellin was detected in the culture medium. Also at least three cytokinin-like substances, equivalent to about 0.001 mug of kinetin per ml, were present. The morphology of pearl millet roots changed when plants in solution culture were inoculated. The number of lateral roots was increased, and all lateral roots were densely covered with root hairs. Experiments with pure plant hormones showed that gibberellin causes increased production of lateral roots. Cytokinin stimulated root hair formation, but reduced lateral root production and elongation of the main root. Combinations of indole acetic acid, gibberellin, and kinetin produced changes in root morphology of pearl millet similar to those produced by inoculation with A. brasilense. PMID- 16345373 TI - Attachment to autoclaved soil of bacterial cells from pure cultures of soil isolates. AB - Pure cultures of Arthrobacter globiformis and four fresh soil isolates were incubated individually in autoclaved soil, in both the presence and absence of glucose. These bacteria grew in the soil and, except for A. globiformis, eventually attached firmly to the soil solids. Firmly attached cells were defined as those which could not be separated from the soil solids by blending combined with a series of low-speed centrifugal washings. The attachment attained by the soil isolates appeared to duplicate that of the overall bacterial population that resides naturally in unaltered, unamended soil. Cell attachment in the autoclaved soil system was accelerated slightly by glucose, but, except for one soil isolate, several months of incubation were still required before firm attachment was complete. Electron microscopy indicated that all attached cells produced extracellular polysaccharide slimes in the autoclaved soil and that these materials appeared to connect the cells to surrounding pieces of soil debris. The actual role of polysaccharides in attachment was not clear, however, because at least one of these organisms possessed extracellular slime during the long period in which it had not yet attached to the soil. PMID- 16345374 TI - Genetic Evidence for Plasmid-Linked Lactose Metabolism in Streptococcus lactis subsp. diacetylactis. AB - It has been previously observed that loss of plasmid pGK4101 occurred concomitantly with loss of lactose-fermenting ability in Streptococcus lactis subsp. diacetylactis 18-16. Transfer of this 41-megadalton plasmid to LM0230, a lactosenegative (Lac) strain of S. lactis, required cell-to-cell contact and resulted in a conversion of LM0230 to the Lac phenotype. This confirms the linkage of lactose-fermenting ability to the 41-megadalton plasmid in S. lactis subsp. diacetylactis and, in addition, demonstrates transfer by a process resembling conjugation in the group N streptococci. PMID- 16345375 TI - Hydrogen Production by the Photosynthetic Bacterium Rhodospirillum rubrum. AB - Continuous photosynthetic production of hydrogen by Rhodospirillum rubrum in batch cultures was observed up to 80 days with the hydrogen donor, pure lactate or lactic acid-containing wastes, supplied periodically. Hydrogen was produced at an average rate of 6 ml/h per g (dry weight) of cells with whey as a hydrogen donor. In continuous cultures with glutamate as a growth-limiting nitrogen source and lactate as a hydrogen donor, hydrogen was evolved at a rate of 20 ml/h per g (dry weight). The composition of the gas evolved remained practically constant (70 to 75% H(2), 25 to 30% CO(2)). Photosynthetic bacteria processing specific organic wastes could be an advantage in large-scale production of hydrogen together with food protein of high value, compared to other biological systems. PMID- 16345376 TI - Heterotrophic uptake experiments with C-labeled histidine in a histidine-limited chemostat. AB - The histidine uptake by bacterial strain HIS 42 was determined with [U C]histidine and through oxygen uptake experiments on samples taken from a histidine-limited chemostat. The uptake of [U-C]histidine was characterized by a saturation constant of 12.8 to 78.6 nM histidine. At higher growth rates, the measured maximum uptake rate of histidine was lower than the actual uptake rate in the culture. The percentage of respired substrate (76 to 93%) was about 30 to 40% higher than the comparable value for the culture. The uptake of histidine as analyzed through the measurement of oxygen uptake rates was characterized by a saturation constant of 1.7 to 10.5 muM histidine; the maximum uptake rate was always greater than the actual histidine uptake rate in the culture. By the application of the two cited methods, set up to determine the histidine uptake kinetics, two different uptake processes were analyzed. It appeared that the determination of the histidine uptake through measurement of the oxygen uptake rate showed a better reflection of the actual uptake process of histidine in the culture. With the available data it was impossible to assess a correlation between the uptake of histidine, as determined with [U-C]histidine, and the actual metabolism of the bacterial population. PMID- 16345377 TI - Growth and Polysaccharide Production by Methylocystis parvus OBBP on Methanol. AB - Methylocystis parvus OBBP, an obligate methylotroph originally isolated as a methane-utilizing bacterium, was cultivated on methanol as a sole source of carbon. After adaptation to high methanol levels, this organism grew on methanol with a maximum specific growth rate of 0.65 h. The pH optimum for growth was between 7 and 9, and the temperature optimum was between 30 and 37 degrees C. Methanol concentrations higher than 5% (by weight) were toxic. Formaldehyde, at a concentration greater than 1 mM, inhibited growth. Formate was neither a substrate nor an inhibitor. An extracellular viscous heteropolysaccharide was produced during growth. The maximum production of the total biomass was 14.5 g (dry weight) per liter of broth. The dried biomass contained 22% (wt/wt) crude protein and 62% (wt/wt) polysaccharide. The main components of the polysaccharide were d-glucose (82%) and l-rhamnose (14%). PMID- 16345378 TI - Frequency of dividing cells, a new approach to the determination of bacterial growth rates in aquatic environments. AB - Frequency of dividing cells is suggested to be an indirect measure of the mean growth rate of an aquatic bacterial community. Seasonal changes in frequency of dividing cells were found which covariated with the bacterial uptake of C-labeled phytoplankton exudates. Batch and continuous culture growth experiments, using brackish water bacteria in pure and mixed enrichment cultures, were performed to establish a relationship between frequency of dividing cells and growth rate. An improved technique for bacterial direct counts, using fluorescent staining and epifluorescence microscopy, is presented. Based on a 6-month survey in a coastal area of the Baltic Sea, the bacterial production in the photic zone is estimated. Compared to the total primary production in the area, the bacterial population during this period utilized approximately 25% of the amount of carbon originally fixed by the primary producers. PMID- 16345379 TI - Nitrogen-fixing (acetylene redution) activity and population of aerobic heterotrophic nitrogen-fixing bacteria associated with wetland rice. AB - Nitrogen-fixing activity associated with different wetland rice varieties was measured at various growth stages by an in situ acetylene reduction method after the activities of blue-green algae (cyanobacteria) in the flood water and on the lower portion of the rice stem were eliminated. Nitrogen-fixing activities associated with rice varieties differed with plant growth stages. The activities increased with plant age, and the maximum was about at heading stage. The nitrogen fixed during the whole cropping period was estimated at 5.9 kg of N per ha for variety IR26 (7 days) and 4.8 kg of N per ha for variety IR36 (95 days). The population of aerobic heterotrophic N(2)-fixing bacteria associated with rice roots and stems was determined by the most-probable-number method, using semisolid glucose-yeast extract and semisolid malate-yeast extract media. The addition of yeast extract to the glucose medium increased the number and activity of aerobic heterotrophic N(2)-fixing bacteria. The glucose-yeast extract medium gave higher counts of aerobic N(2)-fixing bacteria associated with rice roots than did the malate-yeast extract medium, on which Spirillum-like bacteria were usually observed. The lower portion of the rice stem was also inhabited by N(2) fixing bacteria and was an active site of N(2) fixation. PMID- 16345380 TI - Decomposition of blue-green algal (cyanobacterial) blooms in lake mendota, wisconsin. AB - Decomposition of natural populations of Lake Mendota phytoplankton dominated by blue-green algae (cyanobacteria) was monitored by using oxygen uptake and disappearance of chlorophyll, algal volume (fluorescence microscopy), particulate protein, particulate organic carbon, and photosynthetic ability (CO(2) up-take). In some experiments, decomposition of C-labeled axenic cultures of Anabaena sp. was also measured. In addition to decomposition, mineralization of inorganic nitrogen and phosphorus were followed in some experiments. Decomposition could be described as a first-order process, and the rate of decomposition was similar to that found by others using pure cultures of eucaryotic algae. Nitrogen and phosphorus never limited the decomposition process, even when the lake water was severely limited in soluble forms of these nutrients. This suggests that the bacteria responsible for decomposition can obtain all of their key nutrients for growth from the blue-green algal cells. Filtration of lake water through plankton netting that removed up to 90% of the algal biomass usually did not cause a similar decrease in oxygen demand, suggesting that most of the particulate organic matter used for respiration of the decomposing bacteria was in a small particle fraction. Short-term oxygen demand correlated well with the particulate chlorophyll concentration of the sample, and a relationship was derived that could be used to predict community respiration of the lake from chlorophyll concentration. Kinetic analysis showed that not all analyzed components disappeared at the same rate during the decomposition process. The relative rates of decrease of the measured parameters were as follows: photosynthetic ability > algal volume > particulate chlorophyll > particulate protein. Decomposition of C labeled Anabaena occurred at similar rates with aerobic epilimnetic water and with anaerobic sediment, but was considerably slower with anaerobic hypolimnetic water. Of the various genera present in the lake, Aphanizomenon and Anabaena were more sensitive to decomposition than was Microcystis. In addition to providing a general picture of the decomposition process, the present work relates to other work on sedimentation to provide a detailed picture of the fate of blue-green algal biomass in a eutrophic lake ecosystem. PMID- 16345381 TI - Preservation of spores of vesicular-arbuscular endophytes by L-drying. AB - The spores of four species of vesicular-arbuscular endophytes were L-dried at 22 degrees C, and their viability was tested after heating at 80 degrees C for up to 40 min. L-drying of spores in the soil in which they developed was a very effective method of preservation of all spore types examined. Slow L-drying of spores separated from soil and supported on glass fiber filters also gave high viability for spores of some species. A scheme for the long-term preservation of vesicular-arbuscular endophyte spores is proposed. PMID- 16345382 TI - Adenosine 5'-triphosphate flux through the north inlet marsh system. AB - The distribution, fluctuation, and short-term transport of total microbial biomass (measured as adenosine 5'-triphosphate [ATP]) was investigated in a large salt marsh creek. Hourly samples were collected synoptically for 25 h from 10 boats positioned across the 320-m width of the creek. Samples were collected from three depths ranging from 0.2 to 8.0 m. Hourly data obtained from each station were graphed, plotting depth against ATP. Subsequently, interpolated ATP values were generated for every one-tenth depth from the surface to the bottom with the use of an 11-point proportional divider. A total of 2,750 values were generated, and a mean value of 0.865 mg of ATP per m was determined. Maximum levels of ATP were found at high tide and minimal values were found at low tide. The distribution of ATP concentrations was found to be complex, with no suggestion of vertical stratification; however, horizontal divisions were apparent. ATP values corrected for direction of flow or velocity indicated two ebb-directed channels; however, when considered in total, there was a net import of ATP through the interface. The total import of ATP for this 25-h sampling period was calculated to be 3.58 kg, corresponding to a net transport of 39.8 mg of ATP per s through the cross section. Results suggest that detailed characterization of a creek transect in terms of ATP or any similar parameter requires the simultaneous measurements of both the concentration of the parameter in question and the velocity at the time and point from which the sample was taken. PMID- 16345383 TI - Population Densities of Rhizobium japonicum Strain 123 Estimated Directly in Soil and Rhizospheres. AB - Rhizobium japonicum serotype 123 was enumerated in soil and rhizospheres by fluorescent antibody techniques. Counting efficiency was estimated to be about 30%. Indigenous populations of strain 123 ranged from a few hundred to a few thousand per gram of field soil before planting. Rhizosphere effects from field grown soybean plants were modest, reaching a maximum of about 2 x 10 cells of strain 123 per g of inner rhizosphere soil in young (16-day-old) plants. Comparably slight rhizosphere stimulation was observed with field corn. High populations of strain 123 (2 x 10 to 3 x 10 cells per g) were found only in the disintegrating taproot rhizospheres of mature soybeans at harvest, and these populations declined rapidly after harvest. Pot experiments with the same soil provided data similar to those derived from the field experiments. Populations of strain 123 reached a maximum of about 10 cells per g of soybean rhizosphere soil, but most values were lower and were only slightly higher than values in wheat rhizosphere soil. Nitrogen treatments had little effect on strain 123 densities in legume and nonlegume rhizospheres or on the nodulation success of strain 123. No evidence was obtained for the widely accepted theory of specific stimulation, which has been proposed to account for the initiation of the Rhizobium-legume symbiosis. PMID- 16345384 TI - Evaluation of microbiological test kits for hydrocarbon fuel systems. AB - Commercially available kits were tested for their ability to detect bacterial and fungal contamination in hydrocarbon fuel systems. The handling ease of the kits was evaluated, and their sensitivity was compared with that of conventional methods. Most kits in both laboratory and field studies compared well with laboratory methods and were sufficiently sensitive to determine contamination in shipboard fuel tanks. PMID- 16345385 TI - Production of higher alcohols during indonesian tape ketan fermentation. AB - A study was made of the higher alcohols (fusel oils) produced during the Indonesian tape ketan fermentation using Amylomyces rouxii as the principal mold, alone or in combination with yeasts belonging to genera commonly found in the tape ketan fermentation (Endomycopsis, Candida, and Hansenula). Total fusel oils increased with length of fermentation. Fusel oils detected in the product distillate included isobutanol and isoamyl and active amyl alcohols. No n propanol was detected. Isobutanol and isoamyl alcohols were formed in the largest amounts. A. rouxii alone produced nearly the same quantity of fusel oils (total production, 275 mg/liter at 192 h) as it did in combination with Endomycopsis burtonii (total production, 292 mg/liter at 192 h).A. rouxii and Endomycopsis fibuliger produced fusel oils totaling 72 mg/liter at 32 h and 558 mg/liter at 192 h. A. rouxii in combination with Candida yeasts produced somewhat more fusel oils, ranging from 590 to 618 mg/liter at 192 h. A. rouxii in combination with Hansenula yeasts produced the least fusel oils, totaling 143 to 248 mg/liter at 192 h. During the first 36 h, production of fusel oils was higher at 30 and 35 degrees C than at 25 degrees C. At 48 h fusel oil production was slightly higher at 30 degrees C than at 35 degrees C. Beyond 48 h, production of fusel oils was higher at 25 degrees C. A. rouxii in combination with Hansenula anomala and Hansenula subpelliculosa produced considerable ethyl acetate, ranging from 145 to 199 mg/liter at 36 h and 354 to 369 mg/liter at 192 h. PMID- 16345386 TI - Measuring radioactive methane with the liquid scintillation counter. AB - Although a gas proportional counter is the most convenient method of measuring the radioactivity of fixed gases such as methane, it cannot be used when high nonradioactive concentrations of methane are present in the gas phase, due to quenching. If only methane and carbon dioxide are present in radioactive form in the gas phase, a liquid scintillation method for measuring these substances can be used. The procedure is described in detail, and the solubility of methane in liquid scintillation cocktails is determined. PMID- 16345387 TI - Measurement of proteolysis in natural waters. AB - Microbiological proteolysis in Lake Champlain water was measured in situ and in vitro by the spectrophotometric measurement of the rate of release of soluble color from an insoluble azure dye derivative of hide powder. Water samples sterilized by microfiltration were never proteolytic. In situ proteolysis was found to be very dependent upon water temperature (1 to 23 degrees C). No measurable activity was observed below 4 degrees C. The in vitro proteolysis rate at 20 degrees C was found to be 2.3 times the rate at 15 degrees C and 6 times the rate at 10 degrees C. Water taken from beneath the ice-covered lake throughout the winter and tested in the laboratory at 20 degrees C was found to show an increasing proteolytic potential during the winter months. The highest activity was obtained as the ice broke up in early spring. Microbiological proteolysis in water from Burlington Harbor was often four times that found in center lake water. In most experiments proteolysis was inhibited completely by 2 mug of Cu and inhibited 67% by 0.75 mug/ml. Proteolysis was markedly stimulated by 20 to 40 mug of Casitone or Casamino Acids per ml. The predominant bacteria growing in the proteolysis flasks were species of Pseudomonas and Flavobacterium. Pure cultures of Pseudomonas required traces of Casitone, Casamino Acids, or yeast extract for proteolysis of hide powder azure, whereas those of Flavobacterium did not. The requirement could not be met by a mixture of 21 amino acids and eight vitamins. PMID- 16345388 TI - Cytochrome Involvement in Mn(II) Oxidation by Two Marine Bacteria. AB - Two marine, Mn(II)-oxidizing bacterial cultures, BIII 45 and BIII 82, were examined spectrophotometrically at ambient temperature for their cytochrome complements. Membrane preparations from an ethylenediaminetetracetate-lysozyme treatment of 48-h cultures of both strains contained type b, c, and o cytochromes. No evidence for a type a cytochrome was noted. "Periplasmic" fractions of both strains also contained small amounts of cytochrome, including cytochrome o, but "intracellular" fractions did not. Type c cytochrome in membrane preparations of culture BIII 45 was consistently reduced by Mn(II) when the membranes were suspended in the periplasmic fraction of the culture. In the case of culture BIII 82, type c cytochrome in membrane preparations was consistently reduced by Mn(II) when the membranes were suspended in either periplasmic or intracellular fractions of the strain. Although, based on previous inhibitor studies, type b cytochrome was also expected to be reduced by Mn(II), no spectrophotometric evidence for its reduction was found, probably because not enough of it was reduced under the steady-state conditions of the experiments. PMID- 16345389 TI - Phanerochaete chrysosporium beta-Glucosidases: Induction, Cellular Localization, and Physical Characterization. AB - Phanerochaete chrysosporium produces intracellular soluble and particulate beta glucosidases and an extracellular beta-glucosidase. The extracellular enzyme is induced by cellulose but repressed in the presence of glucose. The molecular weight of this enzyme is 90,000. The K(m) for p-nitrophenyl-beta-glucoside is 1.6 x 10 M; the K(i) for glucose, a competitive inhibitor, is 5.0 x 10 M. The K(m) for cellobiose is 5.3 x 10 M. The intracellular soluble enzyme is induced by cellobiose; this induction is prevented by cycloheximide. The presence of 300 mM glucose in the medium, however, had no effect on induction. The K(m) for p nitrophenyl-beta-glucoside is 1.1 x 10 M. The molecular weight of this enzyme is approximately 410,000. Both enzymes have an optimal temperature of 45 degrees C and an E(act) of 9.15 kcal (ca. 3.83 x 10 J). The pH optima, however, were approximately 7.0 and 5.5 for the intracellular and extracellular enzymes, respectively. PMID- 16345390 TI - Effect of bacteria and amoebae on rhizosphere phosphatase activity. AB - The contributions of various components of soil microflora and microfauna to rhizosphere phosphatase activity were determined with hydroponic cultures. Three treatments were employed: (i) plants alone (Bouteloua gracilis (H.B.K.) Lag. ex Steud.) (ii) plants plus bacteria (Pseudomonas sp.), and (iii) plants plus bacteria plus amoebae (Acanthamoeba sp.). No alkaline phosphatase was detected, but an appreciable amount of acid phosphatase activity (120 to 500 nmol of p nitrophenylphosphate hydrolyzed per h per plant) was found in the root culture solutions. The presence of bacteria or bacteria and amoebae increased the amount of acid phosphatase in solution, and properties of additional activity were identical to properties of plant acid phosphatase. The presence of bacteria or bacteria and amoebae increased both solution and root phosphatase activities at most initial phosphate concentrations. PMID- 16345391 TI - Nitrogen requirement of iron-oxidizing thiobacilli for acidic ferric sulfate regeneration. AB - Ammonium was shown to be a limiting nutrient for iron oxidation in cultures of Thiobacillus ferrooxidans. In addition, one strain was also able to assimilate nitrate, but not nitrite, for growth and coupled iron oxidation. Some amino acids (0.5 mM) were tested as a source of nitrogen; none clearly stimulated bacterial activity and inhibition was commonly encountered. Complex nitrogenous compounds were inhibitory at high concentrations (0.1 to 0.5%, wt/vol) and, at low concentrations, some clearly stimulated the bacterial iron oxidation in ammonium limited cultures. Enhancement of iron oxidation by these compounds was also observed in ammonium-unlimited cultures, suggesting their possible role in providing trace nutrients and possibly carbon for the bacteria. PMID- 16345392 TI - Measurement of denitrification in two freshwater sediments by an in situ acetylene inhibition method. AB - An acetylene inhibition method was satisfactorily used for the in situ measurement of denitrification in two sediment-water systems incubated for not more than 22 h. In the presence of added nitrate, denitrification acted as a source of nitrous oxide in a drainage pond, but acted as a sink in its absence. The averaged rates of nitrous oxide accumulation with nitrate enrichment in the absence and presence of acetylene were 0.15 and 0.30 mg of N mh, respectively. Acetylene reduction at an average rate of 0.07 mmol of C(2)H(4) formed mh was simultaneously measured in the absence of added nitrate. In a small eutrophic lake where nitrogen was nonlimiting, the in situ rates of sediment denitrification were 0.09 and 0.11 mg of N mh in the presence and absence of macrophytes, respectively, and no acetylene reduction activity was found. PMID- 16345393 TI - Microbial activity in organic soils as affected by soil depth and crop. AB - The microbial activity of Pahokee muck, a lithic medisaprist, and the effect of various environmental factors, such as position in the profile and type of plant cover, were examined. Catabolic activity for [7-C]salicylic acid, [1,4 C]succinate, and [1,2-C]acetate remained reasonably constant in surface (0 to 10 cm) soil samples from a fallow (bare) field from late in the wet season (May to September) through January. Late in January, the microbial activity toward all three compounds decreased approximately 50%. The microbial activity of the soil decreased with increasing depth of soil. Salicylate catabolism was the most sensitive to increasing moisture deep in the soil profile. At the end of the wet season, a 90% decrease in activity between the surface and the 60- to 70-cm depth occurred. Catabolism of acetate and succinate decreased approximately 75% in the same samples. Little effect of crop was observed. Variation in the microbial activity, as measured by the catabolism of labeled acetate, salicylate, or succinate, was not significant between a sugarcane (Saccharum officinarum L.) field and a fallow field. The activity with acetate was insignificantly different in a St. Augustine grass [Stenotaphrum secundatum (Walt) Kuntz] field, whereas the catabolism of the remaining substrates was elevated in the grass field. These results indicate that the total carbon evolved from the different levels of the soil profile by the microbial community oxidizing the soil organic matter decreased as the depth of the soil column increased. However, correction of the amount of carbon yielded at each level for the bulk density of that level reveals that the microbial contribution to the soil subsidence is approximately equivalent throughout the soil profile above the water table. PMID- 16345394 TI - Extracellular Maltase of Bacillus brevis. AB - Bacillus brevis NRRL B-4389 produced extracellular maltase (alpha-glucosidase; EC 3.2.1.20) only in the presence of short alpha-1,4-glucosidic polymers, such as maltose and maltotriose. An optimum medium was developed; it contained 2.5% maltose, 0.5% nonfat dry milk, 0.4% yeast extract, and 0.01% CaCl(2). The enzyme was produced extracellularly during the logarithmic phase of growth; no cell bound activity was detected at any time. Partial purification of the maltase was accomplished by using diethylaminoethyl cellulose batch adsorption, ammonium sulfate precipitation, and Sephadex G-200 gel filtration. Maltase, isomaltase (oligo-1,6-glucosidase), and glucosyltransferase activities were purified 20.0-, 19.1-, and 11.5-fold, respectively. Some properties of the partially purified maltase were determined: optimum pH, 6.5; optimum temperature, 48 to 50 degrees C; pH stability range, 5.0 to 7.0; temperature stability range, 0 to 50 degrees C; isoelectric point, pH 5.2; and molecular weight, 52,000. The relative rates of hydrolysis of maltose (G(2)), maltotriose (G(3)), G(4), methyl-alpha-d-maltoside, G(40), dextrin, and isomaltose were 100, 22, 12, 10, 10, 8, and 5%, respectively; the K(m) on maltose was 5.8 mM; d-glucose, p-nitrophenyl-alpha-d-glucoside, and tris (hydroxymethyl) aminomethane were competitive inhibitors; transglucosylase activity of the enzyme on maltose resulted in the synthesis of isomaltose, isomaltotroise, and larger oligosaccharides. PMID- 16345395 TI - Growth of Candida ingens on Supernatant from Anaerobically Fermented Pig Waste: Effects of Temperature and pH. AB - Candida ingens, a pellicle-forming yeast utilizing volatile fatty acids, grew over a pH range of 4.1 to 6.0 on nonsterile supernatants from anaerobically fermented pig wastes; growth was inconsistent between pH 4.1 and 4.6. When ambient temperature above the pellicle was 21 degrees C and the temperature of the medium was 29 to 32 degrees C, a pH range of 4.8 to 5.0 gave yields of 1.90 to 3.31 g of dry matter per liter, and 0.059 to 0.065 mol of volatile fatty acids was utilized per liter. There was no advantage in utilization of volatile fatty acids and yield of dry matter in keeping the pH constant during a 24-h growth period. C. ingens grew at pH 4.8 and 5.0 when both ambient and medium temperatures were 30 degrees C. When ambient temperature was 10 degrees C, maximum yield and utilization of volatile fatty acids occurred at a medium temperature of 28 to 30 degrees C. PMID- 16345396 TI - Effect of Ultraviolet-B (280 to 320 nm) Radiation on Blue-Green Algae (Cyanobacteria), Possible Biological Indicators of Stratospheric Ozone Depletion. AB - The effect of ultraviolet-B (280 to 320 nm) irradiation on physiological activities of Anabaena flos-aquae and the water fern Azolla caroliniana has been studied under conditions where lethal effects of irradiation are absent. Nitrogenase activity, measured as acetylene reduction, specifically declined during irradiation with low levels of ultraviolet-B, whereas other physiological activities were unaffected. These findings indicate that measurement of acetylene reduction by cyanobacteria may serve as a specific, sensitive biochemical assay to assess environmental ultraviolet-B effects due to depletion of stratospheric ozone. PMID- 16345397 TI - Na, k, and nonspecific solute requirements for induction and function of galactose active transport in an antarctic psychrophilic marine bacterium. AB - An Antarctic psychrophilic marine Vibrio sp., with the inducible ability to accumulate non-metabolizable [C]methyl-beta-d-thiogalactoside through a galactose transport system, was isolated. Induction of [C]methyl-beta-d-thiogalactoside uptake was found to have a specific Na requirement which was higher than that required for maximal uptake and growth. A specific K requirement was found to be quantitatively the same for uptake, growth, and induction. At low suboptimal growth salinities in artificial seawater, growth, uptake, and induction were inhibited more by the generally low solute concentration than by a specific ion deficiency. Evidence was given that the effect of the nonspecific solute was not completely osmotic in nature. The nonspecific solute requirement was greatest for induction, followed by growth and substrate uptake. PMID- 16345398 TI - Nutritional Studies on Xanthan Production by Xanthomonas campestris NRRL B1459. AB - The nutritional requirements of Xanthomonas campestris NRRL B1459 for optimal xanthan production were studied in a chemically defined medium. Of the carbon sources tested, a 4% sucrose or glucose medium yielded the highest xanthan titers. The further addition of certain organic acids, such as succinate, pyruvate, and alpha-ketoglutarate, stimulated xanthan production; excess concentrations of these organic acids inhibited xanthan formation. Certain amino acids (e.g., glutamate) and nitrate salts were superior to ammonium salts for xanthan production. Concentrations of these nitrogen sources higher than the optimal levels inhibited xanthan production while stimulating growth. Xanthan production was also sensitive to high concentrations of inorganic phosphate. High xanthan potencies, up to 30 g/kg of broth, were achieved in these shake-flask studies, in which completely defined media were used. PMID- 16345399 TI - Plasmid Profiles of Lactose-Negative and Proteinase-Deficient Mutants of Streptococcus lactis C10, ML(3), and M18. AB - Lactose- and proteinase-negative (Lac Prt) mutants of Streptococcus lactis C10, ML3, and M18 were isolated after treatment with ethidium bromide. The Lac Prt mutants of C10 were missing a 40-megadalton plasmid. A 33-megadalton plasmid was absent in the ML3 mutants, and the M18 variants lacked a 45-megadalton plasmid. The results suggest a linkage of these metabolic traits to the respective plasmids. The possible complexity of the interrelationship between lactose metabolism and proteinase activity is presented. PMID- 16345400 TI - Production of Lactase by Candida pseudotropicalis Grown in Whey. AB - Lactase (beta-d-galactosidase) was produced by Candida pseudotropicalis grown in deproteinized whey. Maximum enzyme production in 2% whey was obtained by supplementation with 0.15% yeast extract, 0.1% (NH(4))(2)SO(4), and 0.05% KH(2)PO(4) (wt/vol). Highest enzyme values (4.35 U/mg of cells and 68 U/ml) were obtained with 10 to 12% whey, while enzyme yield was maximal in 2% whey (0.87 U/mg of whey). Optimal initial pH for cultivation was 3.5. The best conditions for extraction included 2% (wt/vol) chloroform, 10 h of treatment, pH 6.6 and higher, and 30 to 37 degrees C. Optimum pH and temperature for enzyme activity were 6.2 and 47 degrees C. The enzyme had a K(m) for O-nitrophenyl-beta-d galactopyranoside of 3.06 x 10 M and the initial V(max) was estimated as 6.63 x 10 M per min. It hydrolized 50 and 100% of the lactose in whey and milk within 4 and 5 h, respectively, at 37 degrees C. The lyophilized enzyme retained 95% of activity for 3 months when stored at -20 degrees C. PMID- 16345401 TI - Cross-Feeding of Lactate Between Streptococcus lactis and Bacteroides sp. Isolated from Termite Hindguts. AB - Streptococcus lactis and Bacteroides sp., isolated from hindguts of Reticulitermes flavipes termites, were grown anaerobically in monoculture and coculture. When grown in a glucose medium, S. lactis monoculture produced lactate as the major fermentation product, with small amounts of formate, acetate, ethanol, and CO(2). In coculture, glucose was completely consumed during growth of S. lactis. Lactate, produced by S. lactis, then supported much of the growth of Bacteroides and was fermented to propionate, acetate, and CO(2). Small amounts of succinate were formed during growth of Bacteroides in the coculture, but little change in the formate or ethanol concentration was observed. Monoculture growth of Bacteroides in a tryptone-yeast extract medium revealed that incorporation of 20 to 40 mM lactate increased cell yields and production of organic acids. However, initial lactate concentrations greater than 40 mM suppressed not only growth of Bacteroides but also acidic product formation. Results suggest that cross-feeding of lactate between streptococci and bacteroides constitutes one aspect of the overall hindgut fermentation in termites. PMID- 16345402 TI - Effect of concentration of organic chemicals on their biodegradation by natural microbial communities. AB - The effect of concentration on the biodegradation of synthetic organic chemicals by natural microbial communities was investigated by adding individual C-labeled organic compounds to stream water at various initial concentrations and measuring the formation of CO(2). The rate of degradation of p-chlorobenzoate and chloroacetate at initial concentrations of 47 pg/ml to 47 mug/ml fell markedly with lower initial concentrations, although half or more of the compound was converted to CO(2) in 8 days or less. On the other hand, little mineralization of 2,4-dichlorophenoxyacetate and 1-naphthyl-N-methylcarbamate, or the naphthol formed from the latter, occurred when these compounds were present at initial concentrations of 2 to 3 ng/ml or less, although 60% or more of the chemical initially present at higher concentrations was converted to CO(2) in 6 days. It is concluded that laboratory tests of biodegradation involving chemical concentrations greater than those in nature may not correctly assess the rate of biodegradation in natural ecosystems and that low substrate concentration may be important in limiting biodegradation in natural waters. PMID- 16345403 TI - The rumen ciliate epidinium in primary degradation of plant tissues. AB - Pieces of lucerne stem suspended in a sheep rumen in nylon bags were removed after different time intervals and examined by scanning electron microscopy. By 15 min large numbers of the ciliate protozoan Epidinium Crawley were attached to damaged areas of the stem, although a complex protozoal fauna was present in the rumen contents. Highest concentrations were on cortex and phloem tissues, with densely packed protozoa forming a complete ring around the transversely cut end of the stem between the epidermis and the vascular cylinder. Within 2 h there was extensive degradation of thin-walled tissues, as indicated by the amount of exposed vascular cylinder. Epidermis was not degraded but slid down the side of the stem as the underlying tissues were removed. This rapid degradation of plant tissue is explained by direct ingestion of tissues by the protozoa on the plant fragments. Many of the epidinia attached to the stem pieces were ingesting phloem elements and chlorophyllous tissue. The rumen protozoa have not previously been shown to participate on this scale in the physical degradation of plant material. PMID- 16345404 TI - Seasonal and geographic distribution of luminous bacteria in the eastern mediterranean sea and the gulf of elat. AB - Luminous bacteria in the Mediterranean Sea and the Gulf of Aqaba-Elat have different distribution patterns. In the Mediterranean Sea, Beneckea harveyi is present all year round, with different subtypes alternating in summer and winter; Photobacterium fischeri was only present during the winter. In the Gulf of Elat, P. leiognathi is present throughout the water column in similar densities during the entire year. This constancy in distribution is presumably due to the near constancy in water temperature. In summer, Photobacterium leiognathi is replaced by B. harveyi in coastal surface waters. In the hypersaline Bardawil lagoon, only B. harveyi types are present. P. fischeri, a major component of the Mediterranean Sea winter communities, is absent from the lagoon. Luminous Beneckea strains show a great diversity in properties, e.g. temperature range for growth, sensitivity to infection by phages, sensitivity to attack by Bdellovibrio strains, and differences in tolerance to high-salinity shock. Therefore, subdivision of the taxonomic cluster of B. harveyi into subtypes is indicated. The composition of the luminous bacteria communities may serve as indicators of different marine water bodies. The symbiotic luminous bacteria of the light organ of the common Gulf of Elat fish, Photoblepharon palbebratus steinitzi, is different from any of the types described. PMID- 16345405 TI - Modified Fluorescent Technique, Using Rhodamine, for Studies of Rhizobium japonicum-Soybean Symbiosis. AB - Rhodamine-conjugated capsular polysaccharides isolated from Rhizobium japonicum 61A76NS were used to examine binding between the labeled polysaccharides and soybean roots. Fluorescein-labeled polysaccharides were not satisfactory because soybean root hairs autofluoresce in the fluorescein region. PMID- 16345406 TI - Oxaloacetate Synthesis in Butyrivibrio fibrisolvens. AB - Phosphoenolpyruvate carboxykinase (adenosine 5'-triphosphate) was the only enzyme capable of carboxylating pyruvate or phosphoenolpyruvate that could be demonstrated in sonicated cells or cell-free extracts of a group 1 butyrivibrio. PMID- 16345407 TI - Examination of methods for enumerating hemicellulose-utilizing bacteria in the rumen. AB - Counts of colonies that developed after 4 days on agar medium containing 0.3% xylan and preincubated rumen fluid were similar to counts of xylanolytic bacteria obtained when total culturable counts were multiplied by the percentage of isolates capable of producing acid from xylan. Shortening the incubation period reduced the chance of including satellite colonies of non-xylanolytic organisms in the count. Nearly all of the xylanolytic isolates irrespective of the medium from which they were isolated degraded and utilized xylan extensively. The use of a culture medium containing a high concentration (3%) of xylan is also described. The number of colonies capable of producing clearings in this medium was less than 10% of the total culturable counts. Isolates from such colonies were shown to produce diffusible (extracellular) xylanases. PMID- 16345408 TI - Rumen anaerobic fungi of cattle and sheep. AB - Plant fragments obtained from natural rumen digesta of fistulated cattle and sheep were examined by scanning electron microscopy. Various plant materials suspended in the rumen for different times were examined likewise. By 2 h large numbers of phycomycetous fungal zoospores were found attached to fibrous plant fragments, particularly vascular tissues. The subsequent development of these fungi resulted in production of thalli with extensive rhizoids and with sporangia up to 175 mum long. Scanning electron microscope examination of plant fragments randomly selected from natural rumen contents of both cattle and sheep demonstrated widespread colonization by large populations of these anaerobic fungi. Furthermore, all plant fragments suspended in nylon bags in the rumen were also extensively colonized. These findings demonstrate that plant fragments in the rumen are the sites of colonization and development by the anaerobic phycomycetous fungi. In addition, the results suggest that these fungi may form a significant part of the rumen microbiota in cattle and sheep fed on fibrous diets and suggest that they may be important in fiber digestion. PMID- 16345409 TI - Isolation of extreme halophiles from seawater. AB - Extreme halophilic bacteria were isolated from the ocean off the coast of Spain. All were gram-negative cocci. One isolate was compared to Halococcus sp. NCMB 757 and was found to have similar characteristics. PMID- 16345410 TI - Improved enumeration of lactic Acid streptococci on elliker agar containing phosphate. AB - Enumeration of lactic acid streptococci from dairy starter cultures on Elliker agar was improved by the addition of 0.4% (wt/vol) diammonium phosphate. Agar plate counts were up to 7.75 times greater due to improved buffering by the phosphate salt. PMID- 16345411 TI - Methane production from cattle waste and delignified straw. AB - Methane production from added substrate per se was approximately twofold greater from delignified straw than that from untreated straw when fermented with cattle manure. PMID- 16345412 TI - Glutamate functions in osmoregulation in a marine bacterium. AB - Beneckea harveyi, growing in either minimal or complex media increased the total amino acid pool with increasing salinity of the medium. Glutamate was the predominate amino acid involved. PMID- 16345413 TI - Development of Defined, Minimal, and Complete Media for the Growth of Hyphomicrobium neptunium. AB - A complete synthetic medium containing 15 amino acids, a minimal synthetic medium (GAMS) containing 4 amino acids, and a supplemented minimal medium (GAMS + calcium pantothenate) have been developed for the cultivation of Hyphomicrobium neptunium ATCC 15444. Depending on the complexity of the synthetic media, generation times were approximately 2 to 3 times longer, and maximum cell densities were 0.3 to 0.9 log(10) lower than in ZoBell marine broth 2216. The fates of C-labeled amino acids in GAMS were monitored. Results suggested that H. neptunium was auxotrophic for methionine, utilized glutamic acid as a primary energy source, and readily anabolized and catabolized serine and aspartic acid. Individual amino acid concentrations above 125 mM induced prolonged lag periods, whereas only methionine was not growth limiting at a concentration as low as 2 mM. PMID- 16345414 TI - Distribution of aerobic bacteria which contain bacteriochlorophyll a. AB - Sixteen strains of aerobic bacteria which contain bacteriochlorophyll a were isolated from the samples collected in aerobic marine environments: thalli of Enteromorpha linza, Porphyra sp., Sargussum horneri; beach sand; and the surface seawater from Aburatsubo Inlet. When they occurred, their proportions among the aerobic heterotrophic populations ranged from 0.9 to 1.1% in the seaweed samples and from 1.2 to 6.3% in the beach sand samples and were 0.9% in the seawater sample. The results suggested that the aerobic photopigmented bacteria widely inhabit aerobic marine environments. PMID- 16345415 TI - Diurnal cycle of oxygen and sulfide microgradients and microbial photosynthesis in a cyanobacterial mat sediment. AB - The diurnal variation in the microgradients of O(2), H(2)S, and Eh were studied in the benthic cyanobacterial mats of a hypersaline desert lake (Solar Lake, Sinai). The results were related to light intensity, light penetration into the mat, temperature, pH, NH(4), photosynthetic activity, pigments, and the zonation of the microbial community. Extreme diurnal variation was found, with an O(2) peak of 0.5 mM at 1 to 2 mm of depth below the mat surface during day and a H(2)S peak of 2.5 mM at 2 to 3 mm of depth at night. At the O(2)-H(2)S interface, the two compounds coexisted over a depth interval of 0.2 to 1 mm and with a turnover time of a few minutes. The photic zone reached 2.5 mm into the mat in summer, and the main CO(2) light fixation took place at 1 to 2 mm of depth. During winter, light and photosynthesis were restricted to the uppermost 1 mm. The quantitative dynamics of O(2) and H(2)S were calculated from the chemical gradients and from the measured diffusion coefficients. PMID- 16345416 TI - Elution and inactivation of bacteriophages on soil and cation-exchange resin. AB - A variety of elution schemes was tested to determine the most effective procedure for eluting Arthrobacter bacteriophages from soil. A buffer solution of pH 8.0 was found to be the most satisfactory eluent. Bacteriophages were adsorbed to cation-exchange sites on soils, clays, and Dowex-50 resin and eluted. Eluted bacteriophages were detected by passive hemagglutination and plaque assay. Although bacteriophage antigen was successfully eluted, most recovered bacteriophages were noninfective. Inactivation was greatest in bacteriophages of Bradley group B and was associated with the elution process. Inactivation is believed to be caused by physical damage to bacteriophage tails upon elution. The significance of this inactivation relative to bacteriophage models for vertebrate viruses is discussed. PMID- 16345417 TI - Continuous Cultivation for Apparent Optimization of Defined Media for Cellulomonas sp. and Bacillus cereus. AB - Steady-state continuous culture was used to optimize lean chemically defined media for a Cellulomonas sp. and Bacillus cereus strain T. Both organisms were extremely sensitive to variations in trace-metal concentrations. However, medium optimization by this technique proved rapid, and multifactor screening was easily conducted by using a minimum of instrumentation. The optimized media supported critical dilution rates of 0.571 and 0.467 h for Cellulomonas and Bacillus, respectively. These values approximated maximum growth rate values observed in batch culture. PMID- 16345418 TI - Effect of monensin and lasalocid-sodium on the growth of methanogenic and rumen saccharolytic bacteria. AB - It is thought that monensin increases the efficiency of feed utilization by cattle by altering the rumen fermentation. We studied the effect of monensin and the related ionophore antibiotic lasalocid-sodium (Hoffman-LaRoche) on the growth of methanogenic and rumen saccharolytic bacteria in a complex medium containing rumen fluid. Ruminococcus albus, Ruminococcus flavefaciens, and Butyrivibrio fibrisolvens were inhibited by 2.5 mug of monensin or lasalocid per ml. Growth of Bacteroides succinogenes and Bacteroides ruminicola was delayed by 2.5 mug of monensin or lasalocid per ml. Populations of B. succinogenes and B. ruminicola that were resistant to 20 mug of either drug per ml were rapidly selected by growth in the presence of each drug at 5.0 mug/ml. Selenomonas ruminantium was insensitive to 40 mug of monensin or lasalocid per ml. Either antibiotic (10 mug/ml) inhibited Methanobacterium MOH, Methanobacterium formicicum, and Methanosarcina barkeri MS. Methanobacterium ruminantium PS was insensitive to 40 mug of monensin or 20 mug of lasalocid per ml. The methanogenic strain 442 was insensitive to 40 mug of monensin but sensitive to 10 mug of lasalocid per ml. The results suggest that monensin or lasalocid acts in the rumen by selecting for succinate-forming Bacteroides and for S. ruminantium, a propionate producer that decarboxylates succinate to propionate. The selection could lead to an increase in rumen propionate formation. Selection against H(2) and formate producers, e.g. R. albus, R. flavefaciens, and B. fibrisolvens, could lead to a depression of methane production in the rumen. PMID- 16345419 TI - Anaerobic biodegradation of eleven aromatic compounds to methane. AB - A range of 11 simple aromatic lignin derivatives are biodegradable to methane and carbon dioxide under strict anaerobic conditions. A serum-bottle modification of the Hungate technique for growing anaerobes was used for methanogenic enrichments on vanillin, vanillic acid, ferulic acid, cinnamic acid, benzoic acid, catechol, protocatechuic acid, phenol, p-hydroxybenzoic acid, syringic acid, and syringaldehyde. Microbial populations acclimated to a particular aromatic substrate can be simultaneously acclimated to other selected aromatic substrates. Carbon balance measurements made on vanillic and ferulic acids indicate that the aromatic ring was cleaved and that the amount of methane produced from these substrates closely agrees with calculated stoichiometric values. These data suggest that more than half of the organic carbon of these aromatic compounds potentially can be converted to methane gas and that this type of methanogenic conversion of simple aromatics may not be uncommon. PMID- 16345420 TI - Attached Growth of Sphaerotilus natans in Continuous-Flow Apparatus and Its Growth Inhibition by 9-beta-d-Arabinofuranosyladenine. AB - Sphaerotilus natans grew at the maximum specific growth rate (mu(max)) of 0.43/h when cultivated on PGY medium at 25 degrees C. The organism mainly grew attached to inside of the culture vessels when the culture medium was fed to the completely mixed continuous-flow apparatus at a dilution rate above mu(max) and the attached growth was directly related to the dilution rate. When a low concentration of the medium was supplied to the apparatus, almost all of the cells grown were filamentous and attached to the inside of the vessels. When a high concentration of the medium was fed, the organism grew as single cells or short chains and flowed out into the effluent. The attached growth of S. natans in the continuous-flow apparatus was inhibited by the minimal inhibitory concentration of 0.5 to 1.0 mug of 9-beta-d-arabinofuranosyladenine per ml. 9 beta-d-Arabinofuranosyladenine showed bacteriocidal activity against S. natans at a concentration of 50 to 100 mug/ml. PMID- 16345421 TI - Automated enzymatic method for the determination of ammonia: application to rumen fluid, gut fluid, and plasma. AB - The enzymatic method for the determination of ammonia with glutamate dehydrogenase has been adapted to the AutoAnalyzer. The automated method was applicable for NH(3) measurement in sheep rumen samples, swine gut fluid samples, and bovine plasma. Results were compared with those of the Conway diffusion method and the manual enzymatic method. The automated method allows 30 samples per h to be analyzed routinely. Values were comparable with those with the manual method, but lower than those with the Conway diffusion method. PMID- 16345422 TI - Rate-temperature curves as an unambiguous indicator of biological activity in soil. AB - Experiments are described in which we used a mass spectrometer to monitor O(2) uptake of enclosed soil samples as a function of temperature. We found that an Arrhenius plot of the rate of O(2) uptake showed pronounced local maxima attributable to biological activity, whereas similar plots of rates obtained with abiotic soils yielded straight lines. This procedure thus provides a basis for distinguishing biological from chemical activity for reactions, such as O(2) uptake, that can occur via either biological or chemical pathways. PMID- 16345423 TI - Biodegradation of C-labeled model and cornstalk lignins, phenols, model phenolase humic polymers, and fungal melanins as influenced by a readily available carbon source and soil. AB - After 6 months of incubation in a fertile neutral sandy loam, about 48% of the ring carbons and 2-carbons and 60% of the OCH(3) carbons of specifically labeled coniferyl alcohol had evolved as CO(2). After 1 year, corresponding values were 55 and 65%. When coniferyl alcohol units were linked into model and cornstalk lignins, about 23% of the ring carbons and 2-carbons and 39% of the OCH(3) carbons had evolved as CO(2) after 6 months. After 1 year, corresponding values were about 28 and 46%. The addition of orange leaves (0.5%, wt/wt) after 6 months did not significantly increase the evolution of CO(2). Addition of orange leaves (0.5%, wt/wt) with specifically C-labeled pyrocatechol, coumaryl alcohol, model lignins, humic acid-type phenolic polymers and of uniformly C-labeled fungal melanins did not increase labeled C losses or C losses from the orange leaves. Decomposition of protein and pyrocatechol linked into model humic acid polymers, coniferyl alcohol C in model lignins, and Eurotium echinulatum melanin in six soils varied from 2 to 14%. Significant differences in C losses were related to soils and were not influenced by orange leaf applications. PMID- 16345424 TI - Production of S-Methylthioacetate by Brevibacterium linens. AB - Volatile sulfur compounds production by eight strains of Brevibacterium linens isolated from cheeses was demonstrated: methanethiol, dimethyldisulfide, and 2,3,4-trithiapentane. Four of these strains also produced S-methylthioacetate, an important aroma component of smear-coated cheeses. It is the first demonstrated microbiological production of a thioester. PMID- 16345425 TI - Arylsulfatase activity in salt marsh soils. AB - The presence of arylsulfatase(s) was confirmed in salt marsh soils. The temperatures of maximum activity and inactivation, the pH range over which the enzyme was active, and the K(m) values were similar to those of soil enzymes. Unlike soil arylsulfatases, however, the salt marsh enzymes do not appear to be repressed by sulfate. It is postulated that these enzymes may be necessary for the initiation of arylsulfate ester metabolism. PMID- 16345426 TI - Muramic Acid assay in sediments. AB - An improved chromatographic assay for muramic acid which is sufficiently sensitive for marine sandy sediments is described; it involves acid hydrolysis, thin-layer chromatography, and gas-liquid chromatography. PMID- 16345427 TI - Carboxymethyl cellulose decomposition by intestinal bacteria of cockroaches. AB - The amounts of carboxymethyl cellulase present in the midguts and hindguts of two species of cockroaches were determined. Numbers of colonies from the guts which digested carboxymethyl cellulose, visualized by flooding plates with cetyltrimethylammonium bromide after incubation, were counted. The numbers indicated the presence of a resident, multiplying population of predominantly anaerobic carboxymethyl cellulose-digesting bacteria in both species of cockroaches. Anaerobic and facultatively anaerobic species of bacteria were isolated, and their carboxymethylcellulase activities were assayed. PMID- 16345428 TI - Comparative study of different iron compounds in inhibition of sphaerotilus growth. AB - The effectiveness of iron compounds on growth inhibition of Sphaerotilus species was compared. In this study, two strains of Sphaerotilus were tested with different iron concentrations in a synthetic sewage (S-medium) as formulated by Lackey and Wattie (Sewage Works J. 12:669-684, 1940). For both strains, >80% inhibition of the maximum respiration rate was obtained by the following levels of soluble iron concentrations at pH 6.0: iron citrate, 20 mg/liter as Fe; iron cysteine, 5 mg/liter as Fe; and ferrous sulfate, 10 mg/liter as Fe. At a pH of 6.7 with iron citrate (20 mg/liter as Fe), inhibition of both strains was in excess of 50%. Insoluble iron compounds, such as iron hydroxides and ferrous carbonate, were found to be much less effective than the soluble iron compounds as inhibitors of these two strains. Aged iron hydroxide (500 mg/liter as Fe) produced a 70% inhibition in the maximum respiration rate while fresh iron hydroxide (52 mg/liter as Fe) and ferrous carbonate (500 mg/liter as Fe) produced a 20% inhibition. Chemical analyses of the iron-inhibited Sphaerotilus strains showed a close relationship between the inhibition of the organism's growth and the amount of iron sorbed by the organism. PMID- 16345429 TI - Luminous enteric bacteria of marine fishes: a study of their distribution, densities, and dispersion. AB - Three taxa of luminous bacteria (Photobacterium fischeri, P. phosphoreum, and Beneckea spp.) were found in the enteric microbial populations of 22 species of surface- and midwater-dwelling fishes. These bacteria often occurred in concentrations ranging between 10 and 10 colony-forming units per ml of enteric contents. By using a genetically marked strain, it was determined that luminous cells entering the fish during ingestion of seawater or contaminated particles traversed the alimentary tract and survived the digestive processes. After excretion, luminous bacteria proliferated extensively on the fecal material and became distributed into the surrounding seawater. Thus, this enteric habitat may serve as an enrichment of viable cells entering the planktonic luminous population. PMID- 16345430 TI - Enumeration and isolation of lactate-utilizing bacteria from the rumen of sheep. AB - A highly specific medium was developed for the enumeration of lactate-utilizing bacteria in the rumen of sheep. This medium, which contained 2.0% lactate, 2.0% Trypticase, 0.2% yeast extract, and volatile fatty acids, hemin, and trace elements in place of rumen fluid, enabled high counts (42 x 10 to 190 x 10/g of ingesta) of lactate-utilizing bacteria to be made with a high degree of specificity (96%). The medium also supported the growth of all species of predominant lactate-utilizing bacteria reported to occur in the rumen and thus is of importance for ecological studies where the incidence and influence of the different species on lactate metabolism under changing conditions in the rumen cannot be predicted. The survival rate of isolates was increased from 60 to 96% by addition to the modified maintenance medium of 40% rumen fluid in place of the volatile fatty acids, hemin, and trace elements used in the counting medium. These results, together with the slow growth of colonies in roll bottles, showed that, although highly selective, the counting medium was not optimal for the types selected. PMID- 16345431 TI - Changes in Lactate-Producing and Lactate-Utilizing Bacteria in Relation to pH in the Rumen of Sheep During Stepwise Adaptation to a High-Concentrate Diet. AB - Changes in the numbers and types of lactate-producing and lactate-utilizing bacteria in the rumen of sheep were followed during stepwise adaptation from a low- to a high-concentrate diet. The mean numbers of bacteria increased after each change in diet when increasing amounts of maize grain were substituted for maize stover. A surge in number of amylolytic bacteria always preceded an increase in lactate-utilizing bacteria, and with the final diet containing 71% grain and molasses the two groups tended to balance each other, which resulted in low lactic acid accumulation. The lactate utilizers thus played a key role in controlling the fermentation. Orderly shifts occurred among the predominating amylolytic and lactate-utilizing bacteria in response to the gradual decrease in ruminal pH as the amount of maize meal in the diet increased. Among the lactate utilizers, the succession began with acid-sensitive Veillonella and Selenomonas, which were superseded by more acid-tolerant Anaerovibrio and Propionibacterium. Among the amylolytic bacteria, Bacteroides was superseded by more acid-tolerant Lactobacillus and Eubacterium. The ecological succession of predominating genera was shown to be correlated significantly with ruminal pH and, more specifically, with the length of time as well as the extent to which the pH remained below a certain critical undefined value in the rumen, arbitrarily set at pH 6.00. PMID- 16345432 TI - Hydrogen Production by the Thermophilic Alga Mastigocladus laminosus: Effects of Nitrogen, Temperature, and Inhibition of Photosynthesis. AB - Hydrogen production by nitrogen-limited cultures of a thermophilic blue-green alga (cyanobacterium), Mastigocladus laminosus, was studied to develop the concept of a high-temperature biophotolysis system. Biophotolytic production of hydrogen by solar radiation was also demonstrated. Hydrogen consumption activity in these cultures was relatively high and is the present limiting factor on both the net rate and duration of hydrogen production. PMID- 16345433 TI - Binding characteristics of n(2)-fixing bacteria to cereal roots. AB - The attachment of Rhizobium japonicum 61A89 and Rhizobium spp. 32H1 to the roots of wheat and rice seedlings is analyzed in terms of an equilibrium model. A Langmuir adsorption isotherm describes the binding. Strain 61A89 binds to a greater extent than does strain 32H1, and the equilibrium constants for each strain binding to wheat are strongly temperature dependent. Both time-dependent dissociation and association, predicted by an equilibrium model, have been found. The dissociation rate constant for 32H1 is approximately twice that of 61A89, and each is weakly temperature dependent. The rate equation for the binding of exponentially growing 61A89 to wheat roots has been solved as a function of time. Theory and experiment both indicate that the binding at very short times is much less than the equilibrium values. The binding of Azotobacter vinelandii 12837 to wheat roots has also been measured. Root-associated Azotobacter fixes nitrogen, whereas under aerobic growth conditions, root-associated 61A89 and 32H1 do not. The effect of metabolic inhibitors and antibiotics on the binding of Rhizobia and Azotobacter was examined. PMID- 16345434 TI - Decomposition of microbial cell components in a semi-arid grassland soil. AB - Cell component fractions (C-labeled) were prepared from bacterial and fungal cultures isolated from the Pawnee National Grassland in northeastern Colorado and tested for seasonal changes in degradability. The decomposition of cell component fractions was monitored from May to December of 1977 and during March of 1978, using soil samples taken at 2- to 3-week intervals. The release of CO(2) from bacterial and fungal cell walls was inversely related (P < 0.01) to the soil moisture content. Except for cytoplasm isolated from an Aspergillus sp., all other cytoplasmic and polysaccharide fractions did not demonstrate a significant relationship between soil moisture and decomposability. In general, bacterial cell walls and polysaccharides were more susceptible to decomposition than fungal cell walls, although the seasonal changes in decomposability for both fractions were similar. These patterns of cell component utilization indicate that the decomposition of cell wall material may be more closely linked, on an inverse basis, to the availability of soil moisture and release of soluble, low-molecular weight organics resulting from primary production events. PMID- 16345435 TI - Denitrification, acetylene reduction, and methane metabolism in lake sediment exposed to acetylene. AB - Samples of sediment from Lake St. George, Ontario, Canada, were incubated in the laboratory under an initially aerobic gas phase and under anaerobic conditions. In the absence of added nitrate (NO(3)) there was O(2)-dependent production of nitrous oxide (N(2)O), which was inhibited by acetylene (C(2)H(2)) and by nitrapyrin, suggesting that coupled nitrification-denitrification was responsible. Denitrification of added NO(3) was almost as rapid under an aerobic gas phase as under anaerobic conditions. The N(2)O that accumulated persisted in the presence of 0.4 atm of C(2)H(2), but was gradually reduced by some sediment samples at lower C(2)H(2) concentrations. Low rates of C(2)H(2) reduction were observed in the dark, were maximal at 0.2 atm of C(2)H(2), and were decreased in the presence of O(2), NO(3), or both. High rates of light-dependent C(2)H(2) reduction occurred under anaerobic conditions. Predictably, methane (CH(4)) production, which occurred only under anaerobiosis, was delayed by added NO(3) and inhibited by C(2)H(2). Consumption of added CH(4) occurred only under aerobic conditions and was inhibited by C(2)H(2). PMID- 16345436 TI - Lytic organisms and photooxidative effects: influence on blue-green algae (cyanobacteria) in lake mendota, wisconsin. AB - The effects of exposure to high light intensities on blue-green algal (cyanobacterial) populations were examined in Lake Mendota, Wis. The algal populations were shown to be susceptible to inhibition of photosynthetic activity and pigment bleaching as a result of exposure. These effects generally influence only a small percentage of the lake population and thus are probably not important in causing major declines in chlorophyll a. Lytic organisms were shown to increase in numbers in the lake in response to the seasonal development of blue-green algae, reaching values of greater than 1,000 plaque-forming units per ml in midsummer. Both bacteria and protozoa were observed in plaque zones, but it could not be determined whether these lytic organisms had a major role in algal biomass declines. PMID- 16345437 TI - Effects of Ammonium Ions, Oxygen, Carbon Monoxide, and Acetylene on Anaerobic and Aerobic Hydrogen Formation by Anabaena cylindrica B629. AB - An investigation was made of various factors, both experimental and physiological, which influenced the formation of hydrogen gas by the heterocystous cyanobacterium Anabaena cylindrica B629 when incubated in both argon and air. A. cylindrica B629 produces hydrogen in air in the presence of carbon monoxide, acetylene, or both, with a short lag period. The rate of production in air at optimal concentrations of these compounds was found to be comparable with that in an argon atmosphere. Whereas under argon, ammonium ions (0.5 to 6 mM) were found to inhibit hydrogen formation in a manner which was dependent on light intensity and not relieved by oxygen (1 to 20% of gas phase), in air-carbon monoxide-acetylene, these ions (up to at least 0.5 mM) slightly stimulated hydrogen production for at least 24 h. Conclusions are drawn about short-term aerobic and anaerobic hydrogen formation by A. cylindrica B629 and the effects of ammonium ions, oxygen, carbon monoxide, and acetylene on these processes. PMID- 16345438 TI - Duration of Hydrogen Formation by Anabaena cylindrica B629 in Atmospheres of Argon, Air, and Nitrogen. AB - The time course of hydrogen formation by Anabaena cylindrica was followed beneath an argon atmosphere alone and also beneath atmospheres of argon, nitrogen, and air in the presence of carbon monoxide (0.2%) and acetylene (5%). Hydrogen production beneath argon alone was comparable in rate and duration (7 to 12 days) to that which occurred beneath air in the presence of carbon monoxide (0.2%) and acetylene (5%). However, much greater longevity (16 to 26 days) and improved rates of hydrogen formation were obtained when algae were incubated beneath argon and particularly nitrogen, each supplemented with carbon monoxide and acetylene. The total hydrogen produced by these cultures was up to three times as much as that released by cultures incubated beneath argon alone. Hydrogen-oxygen ratios for argon cultures either with or without carbon monoxide and acetylene were initially 1:5 but approximated 1:2 when measured over the entire incubation period. In each case oxygen production and nitrogenase activity (acetylene reduction) continued at reduced rates after hydrogen evolution had ceased. The effects of methionine sulfoximine (2 muM), ammonium ions (0.5 mM), or both on oxygen production were generally negligible, while effects on hydrogen production were variable depending on the atmosphere used; in most cases, eventual destabilization of the system occurred. A brief comparison was made of the time courses of anaerobic and aerobic hydrogen formation by the marine cyanobacterium Calothrix membranacea. It was found that shaking of cultures was beneficial for hydrogen production but not strictly necessary. It is concluded that hydrogen production by A. cylindrica in air and particularly nitrogen in the presence of carbon monoxide and acetylene offers the best potential of the atmospheres considered on the basis of four criteria: rates and longevity of hydrogen formation, practicality of the atmosphere used, and tolerance of hydrogen evolution to slight changes in composition of the atmosphere. PMID- 16345439 TI - General Properties of Beta-Galactosidase of Xanthomonas campestris. AB - Partially purified beta-galactosidase of Xanthomonas campestris required 32 to 37 degrees C and pH 5.5 to 5.8 for optimum activity. The enzyme had low affinity for lactose hydrolysis (K(m) = 22 mM) and was inhibited by thiol group reagents, ethylenediaminetetraacetic acid, galactose, and d-galactal. PMID- 16345440 TI - Degradation of poliovirus by adsorption on inorganic surfaces. AB - [This corrects the article on p. 483 in vol. 37.]. PMID- 16345441 TI - Algae/Bacteria ratio in high-rate ponds used for waste treatment. AB - Algae, bacteria, and zooplankton were counted in samples drawn from 120- and 150 m high-rate algae ponds (those used for wastewater treatment). The fraction of nondegraded organic matter was estimated by comparing the ratio of biological and chemical oxygen demands and the bacterial, algal, and zooplankton counts to volatile suspended solids. With pond effluent quality at an acceptable level (around 18 mg of dissolved biological oxygen demand), the algae/bacteria ratio was around 1:100 or even higher, the zooplankton count was negligible, and the bacterial concentration was approximately 10 cells per liter by direct count. The data for bacteria exceeded those of earlier studies by one to three orders of magnitude. PMID- 16345442 TI - Physiological characteristics underlying the distribution patterns of luminous bacteria in the mediterranean sea and the gulf of elat. AB - Physiological characteristics of luminous bacteria isolated from the Mediterranean and Gulf of Elat were compared to determine their relationship to the specific seasonal and geographic distribution patterns of these bacteria. The effects of temperature on growth rate and yield, relative sensitivity to photooxidation, resistance to high salt concentration (8%), and ability to grow in nutrient-poor conditions appear to control these patterns. The winter appearance of Photobacterium fischeri and the succession of winter and summer types of Beneckea harveyi in the eastern Mediterranean are explained by different temperature requirements for growth. Sensitivity to photooxidation explains the disappearance of P. leiognathi, present in the main body of the Gulf of Elat throughout the year, from the shallow coastal strip. B. harveyi is present in this coastal strip which is higher in nutrients and in productivity than the open waters. Competition experiments between B. harveyi and P. leiognathi in batch and continuous culture indicate that the oligotrophic P. leiognathi is outcompeted by B. harveyi in rich and even in relatively poor media. The distribution pattern found in the Bardawil hypersaline lagoon is explained by selection of salinity resistant mutants of B. harveyi from the Mediterranean Sea. PMID- 16345443 TI - Survival of Rhizobium phaseoli in Coal-Based Legume Inoculants. AB - The long-term survival of Rhizobium phaseoli strains 127K17, 127K26, and 127K35 in legume inoculants prepared with eight different coals (one strain and one coal per inoculant) was studied. The coals used were Pennsylvania anthracite, bituminous coals from Illinois, Pennsylvania, and Utah, lignite from North Dakota and Texas, and subbituminous coals from New Mexico and Wyoming; they ranged in pH from 4.7 to 7.5 All coals, with the exceptions of Illinois bituminous coal and Texas lignite (pH's of 5.0 and 4.7, respectively), supported the growth and survival of all R. phaseoli strains. All coal-based inoculants in which rhizobial viability was maintained had more than 10 rhizobia per g for at least 7 months, and most contained more than 10 rhizobia per g after 12 months. It appears that most coals, regardless of grade or source, may be acceptable carriers for R. phaseoli inoculants. PMID- 16345444 TI - Purification and Some Properties of an Extracellular Amylase from a Moderate Halophile, Micrococcus halobius. AB - A moderate halophile, Micrococcus halobius ATCC 21727, produced an extracellular dextrinogenic amylase when cultivated in media containing 1 to 3 M NaCl. The amylase was purified from the culture filtrate to an electrophoretically homogenous state by glycogen-complex formation, diethylaminoethyl-cellulose chromatography, and Bio-Gel P-200 gel filtration. The enzyme had maximal activity at pH 6 to 7 in 0.25 M NaCl or 0.75 M KCl at 50 to 55 degrees C. The activity was lost by dialysis against distilled water. Molecular weight was estimated to be 89,000 by sodium dodecyl sulfate-gel electrophoresis. The action pattern on amylose, soluble starch, and glycogen showed that the products were maltose, maltotriose, and maltotetraose, with lesser amount of glucose. PMID- 16345445 TI - Effect of inoculation with n(2)-fixing spirilla and azotobacter on nitrogenase activity on roots of maize grown under subtropical conditions. AB - Inoculated and non-inoculated seedlings of maize were grown in fertile clayloam soils of Egypt and Belgium under subtropical conditions provided in a greenhouse. Acetylene-reducing activity and microbial counts were determined during a period ranging from 6 to 12 weeks after sowing. Irrespective of soil origin, N(2)-fixing spirilla and Azotobacter were common under maize cultivation. Inoculation resulted in a transitional increase in their numbers at early stages of growth. Nitrogenase activity was not detected in the rhizosphere of young plants. The maximum activities measured (81 to 1,436 nmol of C(2)H(4) g h) occurred close to the 50 to 70% silking stage. Inoculation with N(2)-fixing spirilla, particularly in Nile Delta soil, doubled the amount of N(2) fixed in a late period of growth (12 weeks), whereas inoculation with Azotobacter had no noticeable effect. PMID- 16345446 TI - Extracellular Polysaccharide of Erwinia amylovora: a Correlation with Virulence. AB - The extracellular polysaccharides produced as slime or capsule layers by bacterial pathogens of animals and plants have been often implicated as factors essential to pathogenesis. In the present study, virulence of the plant pathogen Erwinia amylovora was correlated with the ability to produce extracellular polysaccharide (EPS). EPS production by a series of field isolates and bacterio phage-resistant mutants differing in the extent to which they cause symptoms in host tissues was examined by quantitation with a modified Laurell rocket immunoelectrophoresis assay. The amount of EPS produced as an easily removed capsular layer or slime on solid nutrient agar approximated the capacity to exhibit symptoms in host inoculation tests. Features common to the virulent isolates are mucoid colony morphology, sensitivity to EPS-specific bacteriophages [Sphi3 and PEal(h)], and ability to produce a characteristic EPS. Mutants selected for resistance to Sphi3 or nonmucoid colony morphology are deficient in EPS production and have lost the ability to multiply in host tissue and cause symptoms. We conclude that EPS may be directly involved in symptom expression and provide a function essential to the growth of the pathogen in host tissues. PMID- 16345447 TI - Temporal change in nitrous oxide and dinitrogen from denitrification following onset of anaerobiosis. AB - Similar temporal patterns were found in three mineral soils for the composition of the gaseous products of denitrification following the onset of anaerobic conditions. During the early period of anaerobiosis (0 up to 1 to 3 h), N(2) was the dominant product of denitrification. The NO(3) --> N(2)O activity then increased, but was not accompanied by a corresponding increase in N(2)O-reducing activity. This resulted in a relatively extended period of time (1 to 3 up to 16 to 33 h) during which N(2)O was a major product. Eventually (after 16 to 33 h), an increase in N(2)O-reducing activity occurred without a comparable increase in the N(2)O-producing activity. The increase in the rate of N(2)O reduction did not occur in the presence of chloramphenicol and required the presence of N(2)O or NO(3) during the preceding anaerobic incubation. During the final period (16 to 33, up to 48 h), N(2) was generally the sole product of denitrification, since the rate of N(2)O reduction exceeded the rate of N(2)O production. A similar sequential pattern was also found for a culture of a denitrifying Flavobacterium sp. shifted to anaerobic growth. A staggered synthesis of the enzymes in the denitrification sequence apparently occurred in response to anoxia, which caused first a net production of N(2)O followed by consumption of N(2)O. PMID- 16345448 TI - Maceration of Clover and Grass Leaves by Lachnospira multiparus. AB - A strain of Lachnospira multiparus, a pectin-hydrolyzing bacterium from the rumen, was incubated in nutrient media in the presence of surface-disinfected clover leaflets. When the culture flasks containing the leaflets together with Lachnospira were shaken after overnight incubation, extensive maceration of the leaflets was seen, although uninoculated control leaflets remained intact during a similar treatment. Examination of inoculated leaflets by transmission electron microscopy showed extensive invasion of intercellular areas of the mesophyll tissue but only minor invasion of vascular tissue. Cutting the leaves before incubation greatly increased the ability of L. multiparus to colonize and macerate the leaflets. Similar experiments with grass leaves are also described, and the possible role of maceration in the digestion of plant material in the rumen is discussed. Although Lachnospira stains gram variable and often gram negative, the ultrastructure of the cell wall was that of a gram-positive bacterium. PMID- 16345449 TI - Enhanced Cellulase Production by a Mutant of Sclerotium rolfsii. AB - A mutant of Sclerotium rolfsii CPC 142 that secretes about two times more filter paper-degrading activity in NM-2 growth medium in submerged cultures than the parent strain was obtained by ultraviolet mutagenesis of crushed sclerotia. The production of endo-beta-glucanase in the mutant was affected to a lesser extent. With the parent strain, the addition of 3% rice bran to NM-2 medium was essential for optimal formation of cellulase, including filter paper-degrading activity. However, with the mutant the addition of rice bran to NM-2 medium increased the formation of endo-beta-glucanase but not filter paper-degrading or cellobiase activity. An altered control mechanism for the production of filter paper degrading enzymes is suggested. The genome(s) controlling the cellulase complex of enzymes in the UV-8 mutant is not under coordinate control. PMID- 16345450 TI - Assessment of microbial fouling in an ocean thermal energy conversion experiment. AB - A project to investigate biofouling, under conditions relevant to ocean thermal energy conversion heat exchangers, was conducted during July through September 1977 at a site about 13 km north of St. Croix (U.S. Virgin Islands). Seawater was drawn from a depth of 20 m, within the surface mixed layer, through aluminum pipes (2.6 m long, 2.5-cm internal diameter) at flow velocities of about 0.9 and 1.8 m/s. The temperature of the seawater entering the mock heat exchanger units was between 27.8 and 28.6 degrees C. After about 10 weeks of exposure to seawater, when their thermal conductivity was reported to be significantly impaired, the pipes were assayed for the accumulation of biological material on their inner surfaces. The extent of biofouling was very low and independent of flow velocity. Bacterial populations, determined from plate counts, were about 10 cells per cm. The ranges of mean areal densities for other biological components were: organic carbon, 18 to 27 mug/cm; organic nitrogen, 1.5 to 3.0 mug/cm; adenosine 5'-triphosphate, 4 to 28 ng/cm; carbohydrate (as glucose in the phenol assay), 3.8 to 7.0 mug/cm; chlorophyll a, 0.2 to 0.8 ng/cm. It was estimated from the adenosine 5'-triphosphate and nitrogen contents that the layer of live bacteria present after 10 weeks was only of the order of 1mum thick. The C/N ratio of the biological material suggested the presence of extracellular polysaccharidic material. Such compounds, because of their water-retaining capacities, could account for the related increase in thermal resistance associated with the pipes. This possibility merits further investigation, but the current results emphasize the minor degree of biofouling which is likely to be permissible in ocean thermal energy conversion heat exchangers. PMID- 16345451 TI - Role of the Salt Marsh Grass Spartina alterniflora in the Response of Soil Denitrifying Bacteria to Glucose Enrichment. AB - Long-term incubations of salt marsh soil systems in the presence of glucose resulted in a decrease in the soils' denitrification potential. Addition of nitrate or the presence of living Spartina alterniflora reversed this effect, indicating that Spartina, through the establishment of an oxidized rhizosphere where nitrification can occur, enables the denitrifying bacteria to adequately compete with the less energetically efficient components of the anaerobic soil microbial community. PMID- 16345452 TI - Calculation of k and v from substrate concentration versus time plot. AB - A simple method for the calculation of kinetic parameters (K(m), V(max)) under conditions of changing substrate concentrations is presented. An application of the method to detect shifts in groups involved in the utilization of a substrate in a mixed microbial culture is given. PMID- 16345453 TI - Agar slush as a method to insure homogeneity of monocentric fungi in continuous culture. AB - Agar granules suspended in the growth medium reduce clumping, which permits the continuous culture of monocentric chytridiomycete fungi. PMID- 16345454 TI - Oxaloacetate Synthesis in Butyrivibrio fibrisolvens. AB - [This corrects the article on p. 1247 in vol. 37.]. PMID- 16345455 TI - Two improved methods for obtaining axenic cultures of cyanobacteria. AB - Scoring the agar plate before incubation under unidirectional light led to a rapid separation of gliding filamentous cyanobacteria from their contaminating bacteria. Twenty strains were purified by the method. Additionally, 13 axenic cyanobacterial strains were isolated from pour plates made after treatment of cyanobacterial cultures in tryptone-yeast extract-glucose broth with cycloserine in darkness to select for obligate photoautotrophs. PMID- 16345456 TI - Isolation of cis-3-Amino-l-Proline from Cultured Mycelia of Morchella esculenta Fr. AB - cis-3-Amino-l-proline, identified once as a nonprotein amino acid from the fruiting bodies of Morchella esculenta Fr., was isolated also from the growth medium and cultured mycelia of the same fungus. PMID- 16345457 TI - Apparatus for monitoring the mineralization of volatile C-labeled compounds. AB - Quantitative mineralization studies on radiolabeled compounds having high vapor pressures need to cope with several technical difficulties. An incubation and trapping system is described that was successfully used in mineralization studies on highly volatile trichlorobenzenes and other xenobiotic pollutants. PMID- 16345458 TI - Continuous Removal of Lactic Acid from Wastewater by Candida utilis. AB - Under steady-state conditions, the maximum rate of lactic acid removal was found to be 588 mg/h per liter of continuous culture. PMID- 16345459 TI - Autotrophic Growth of Thiobacillus acidophilus in the Presence of a Surface Active Agent, Tween 80. AB - Cellular protein, pH, dissolved oxygen concentration, and static surface tension were measured during growth of Thiobacillus acidophilus on elemental sulfur in the absence and presence of up to 5,000 mg of Tween 80 per liter. The decrease in pH and the increase in sulfate production were observed to be less accurate measurements of growth when compared with the increase in cellular protein. The doubling time of the bacterium decreased approximately 50% with the addition of 500 mg of Tween 80 per liter. The bacteria did not appear to synthesize any wetting agents as demonstrated by the constant surface tension of the medium during growth. Morphological alterations in the presence of Tween 80 were also observed. PMID- 16345460 TI - Nutritional Requirements of Microbacterium thermosphactum. AB - Microbacterium thermosphactum requires cysteine, alpha-lipoate, nicotinate, pantothenate, p-aminobenzoate, biotin, and thiamin for aerobic growth in glucose mineral salts medium. Glucose cannot be replaced by Casamino Acids or acids of the tricarboxylic acid cycle as sole carbon and energy sources. The organism can also grow anaerobically in the minimal synthetic medium. PMID- 16345461 TI - Measurement of microbial activity and growth in the ocean by rates of stable ribonucleic Acid synthesis. AB - A relatively simple and extremely sensitive technique for measuring rates of stable ribonucleic acid (RNA) synthesis was devised and applied to bacterial cultures and seawater samples. The procedure is based upon the uptake and incorporation of exogenous radiolabeled adenine into cellular RNA. To calculate absolute rates of synthesis, measurements of the specific radioactivity of the intracellular adenosine 5'-triphosphate pools (precursor to RNA) and of the total amount of radioactivity incorporated into stable cellular RNA per unit time are required. Since the rate of RNA synthesis is positively correlated with growth rate, measurements of RNA synthesis should be extremely useful for estimating and comparing the productivities of microbial assemblages in nature. Adenosine 5' triphosphate, adenylate energy charge, and rates of stable RNA synthesis have been measured at a station located in the Columbian Basin of the Caribbean Sea. A subsurface peak in RNA synthesis (and therefore growth) was located within the dissolved oxygen minimum zone (450 m), suggesting in situ microbiological utilization of dissolved molecular oxygen. Calculations of the specific rates of RNA synthesis (i.e., RNA synthesis per unit of biomass) revealed that the middepth maximum corresponded to the highest specific rate of growth (420 pmol of adenine incorporated into RNA.day) of all depths sampled, including the euphotic zone. The existence of an intermediate depth zone of active microbial growth may be an important site for nutrient regeneration and may serve as a source of reduced carbon for mesopelagic and deep sea environments. PMID- 16345462 TI - Steady-State Growth and Chemical Composition of the Marine Chlorophyte Dunaliella tertiolecta in Nitrogen-Limited Continuous Cultures. AB - The marine chlorophyte Dunaliella tertiolecta was grown in continuous cultures under NH(4)-N, NO(2)-N, NO(3)-N, and urea-N limitations. The effect of the nitrogen cell quota (Q(n)) on the steady-state growth rate (mu) was the same regardless of the N source. The relationship between mu and Q(n) was well described by the Droop equation, but only up to the true maximum growth rate ;mu (= cell washout rate). The ratio between the minimum cell quota (k(Q)) and the maximum cell quota (Q(m)) was 0.19. Hence, there is no substitute for determining ;mu experimentally. That there was no difference in growth response to different N sources suggests that no internal pooling of inorganic nitrogen occurred. Both the carbon (Q(c)) and phosphorus (Q(p)) cell quotas under N limitation increased with increasing mu in a threshold fashion: virtually no change in either cell quota up to approximately 0.8 ;mu, followed by a rapid and large increase up to ;mu. In addition, in the region of low mu, there was an increase in Q(p) with a decreasing medium N/P ratio of between 15 and 5 (by atoms). The results generally indicate the physiological limits in cellular constituency under N limitation. The usefulness of this information, however, in describing the response of natural populations of marine phytoplankton to transient nutrient exposures on the temporal and spatial microscales that most likely exist is of limited value. PMID- 16345463 TI - Effect of interactions among algae on nitrogen fixation by blue-green algae (cyanobacteria) in flooded soils. AB - Nitrogen fixation (C(2)H(2) reduction) by algae in flooded soil was limited by interactions within the algal community. Nitrogen fixation by either indigenous algae or Tolypothrix tenuis was reduced severalfold by a dense suspension of the green alga Nephrocytium sp. Similarly, interactions between the nitrogen-fixing alga (cyanobacterium) Aulosira 68 and natural densities of indigenous algae limited nitrogen-fixing activity in one of two soils examined. This was demonstrated by developing a variant of Aulosira 68 that was resistant to the herbicide simetryne at concentrations that prevented development of indigenous algae. More nitrogen was fixed by the resistant variant in flooded soil containing herbicide than was fixed in herbicide-free soil by either the indigenous algae or indigenous algae plus the parent strain of Aulosira. Interference from indigenous algae may hamper the development of nitrogen-fixing algae introduced into rice fields in attempts to increase biological nitrogen fixation. PMID- 16345464 TI - Alkaline phosphatase assay for freshwater sediments: application to perturbed sediment systems. AB - The p-nitrophenyl phosphate hydrolysis-phosphatase assay was modified for use in freshwater sediment. Laboratory studies indicated that the recovery of purified alkaline phosphatase activity was 100% efficient in sterile freshwater sediments when optimized incubation and sonication conditions were used. Field studies of diverse freshwater sediments demonstrated the potential use of this assay for determining stream perturbation. Significant correlations between phosphatase and total viable cell counts, as well as adenosine triphosphate biomass, suggested that alkaline phosphatase activity has utility as an indicator of microbial population density and biomass in freshwater sediments. PMID- 16345465 TI - Distribution of the Luminous Bacterium Beneckea harveyi in a Semitropical Estuarine Environment. AB - Bioluminescent bacteria were found in the water column, sediment, shrimp, and gastrointestinal tract of marine fishes from the semitropical estuarine environment of the East Lagoon, Galveston Island, Tex. Populations in the water column decreased during cold weather while sedimentary populations persisted. The highest percentages of luminous organisms were isolated from the gastrointestinal tract of marine fishes, where they persisted during 5 days of starvation. The presence of chitin temporarily increased intestinal populations. All isolates were Beneckea harveyi, whose natural habitat appears to be the gut of fishes and whose free-living reservoir appears to be marine sediments. PMID- 16345466 TI - Toxicity of NO(2): Effect of Nitrite on Microbial Activity in an Acid Soil. AB - In an acid forest soil of pH 4.0 to 4.2 amended with glucose, 1.0 mug of nitrite N per g of soil inhibited the rate of O(2) utilization and CO(2) evolution. The inhibition was evident only for several hours after nitrite addition, and the subsequent rate of glucose mineralization was the same as in soil not receiving nitrite. The decomposition of protein hydrolysate was reduced by 10 mug of nitrite-N per g of soil but not lower concentrations, and the inhibition of this process by 20 mug of nitrite-N per g had dissipated after 24 h. Nitrite disappeared readily from this soil. More than 20 mug of bisulfite-S per g of soil was required to inhibit glucose decomposition. The data suggest that the possible antimicrobial effects of low levels of NO(2), which give rise to nitrite in soil, require further evaluation. PMID- 16345467 TI - Influence of substrate composition on marine microfouling. AB - Glass and metal substrates exposed to subtropical seawater exhibit a characteristic succession of periphytic microorganisms. Substrates which are biologically and physically inert (glass and stainless steel) fouled rapidly and produced a complex, two-tier microfouling layer. Characteristic microorganisms were bacteria, fungi, choanoflagellates, diatoms, ciliates, and microalgae. Active substrates (brass and copper-nickel alloys) were selective for bacteria which secrete extracellular mucoid material, fouled at a slower rate, and were characterized by a less diverse microfouling community. Substrate appears to influence microfouling at all stages of development, especially in the early stages of substrate conditioning, and primary film formation. Primary films may insulate periphytic microorganisms from toxic substrates. PMID- 16345468 TI - Isolation and Characterization of a Thermophilic Strain of Methanosarcina Unable to Use H(2)-CO(2) for Methanogenesis. AB - A thermophilic strain of Methanosarcina, designated Methanosarcina strain TM-1, was isolated from a laboratory-scale 55 degrees C anaerobic sludge digestor by the Hungate roll-tube technique. Penicillin and d-cycloserine, inhibitors of peptidoglycan synthesis, were used as selective agents to eliminate contaminating non-methanogens. Methanosarcina strain TM-1 had a temperature optimum for methanogenesis near 50 degrees C and grew at 55 degrees C but not at 60 degrees C. Substrates used for methanogenesis and growth by Methanosarcina strain TM-1 were acetate (12-h doubling time), methanol (7- to 10-h doubling time), methanol acetate mixtures (5-h doubling time), methylamine, and trimethylamine. When radioactively labeled acetate was the sole methanogenic substrate added to the growth medium, it was predominantly split to methane and carbon dioxide. When methanol was also present in the medium, the metabolism of acetate shifted to its oxidation and incorporation into cell material. Electrons derived from acetate oxidation apparently were used to reduce methanol. H(2)-CO(2) was not used for growth and methanogenesis by Methanosarcina strain TM-1. When presented with both H(2)-CO(2) and methanol, Methanosarcina strain TM-1 was capable of limited hydrogen metabolism during growth on methanol, but hydrogen metabolism ceased once the methanol was depleted. Methanosarcina strain TM-1 required a growth factor (or growth factors) present in the supernatant of anaerobic digestor sludge. Growth factor requirements and the inability to use H(2)-CO(2) are characteristics not found in other described Methanosarcina strains. The high numbers of Methanosarcina-like clumps in sludges from thermophilic digestors and the fast generation times reported here for Methanosarcina TM-1 indicate that Methanosarcina may play an important role in thermophilic methanogenesis. PMID- 16345469 TI - Liquid chromatographic determination of dipicolinic Acid from bacterial spores. AB - Dipicolinic acid was determined by reverse-phase liquid chromatography. Elution was with 0.2 M potassium phosphate, pH 1.8, containing 1.5% tert-amyl alcohol or higher concentrations of lower alcohols or acetonitrile. The normal analytical range was 50 to 1,000 muM, which is equivalent to 0.1 to 1 mg of spores per ml with a relative standard error of 2 to 4% and a detection limit of <100 pmol. Dipicolinic acid was fully extracted from spores by heating at pH 1.8 for 10 min at 100 degrees C. Sporulating cultures may be analyzed in less than 20 min without separation of cells from media. Liquid chromatography was also used to detect dipicolinic acid in more complex substrates, e.g., guinea pig feces containing Metabacterium polyspora spores and canned food. Dipicolinic acid could be detected in unspoiled canned salmon containing <10 added Bacillus cereus spores per g. PMID- 16345470 TI - Variability of Temperature, pH, and Moisture in an Aerobic Composting Process. AB - This study measured the environmental variability which exists in a commercial aerobic composting process. The specific process studied is carried out in six decomposition cells which present six different phases of the process. Temperature, pH, and moisture content were determined in several randomly chosen sample sites in each cell, both at the beginning and at the end of the time the material was left in the cell. Temperature and pH varied greatly from one sample site to another in each cell, whereas moisture content was less varied. A significant rise in both temperature and pH was observed at two stages of degradation. PMID- 16345471 TI - Conversion of Glycerol to Dihydroxyacetone by Immobilized Whole Cells of Acetobacter xylinum. AB - Enzymatic production of dihydroxyacetone (DHA) was studied by immobilization of the whole cells of acetic acid bacteria capable of oxidizing glycerol to DHA. Acetobacter xylinum A-9 cells immobilized in a polyacrylamide gel were selected as the most favorable enzyme preparation. The enzymatic properties of immobilized cells converting glycerol to DHA were investigated and compared with those of intact cells. The optimum pH for the immobilized cells was broad (4.0 to 5.5), whereas the intact cells had a narrow pH optimum at 5.5. The thermal stability of the immobilized cells was somewhat higher than that of the intact cells. Apparent K(m) values for glycerol with both intact and immobilized cells were about equal, 6.3 x 10 to 6.5 x 10 M. The complete conversion of glycerol to DHA was achieved within 40 h under optimum conditions, and pure crystalline DHA was readily isolated from the reaction mixture with over 80% yield. PMID- 16345472 TI - Neutron activation analysis for the demonstration of amphibolite rock-weathering activity of a yeast. AB - Neutron activation analysis was employed in a survey of weathering abilities of rock surface microorganisms. A yeast isolated from an amphibolite of a megalithic grave was found actively to concentrate, in media and in or on cells, iron and other elements when grown in the presence of ground rock. This was demonstrated by comparing a spectrum of neutron-activated amphibolite powder (particle size, 50 to 100 mum) with the spectra of neutron-activated, lyophilized yeast cells which had grown with or without amphibolite powder added to different media. The most active yeast (IFAM 1171) did not only solubilize Fe from the rock powder, but significant amounts of Co, Eu, Yb, Ca, Ba, Sc, Lu, Cr, Th, and U were also mobilized. The latter two elements occurred as natural radioactive isotopes in this amphibolite. When the yeast cells were grown with neutron-activated amphibolite, the cells contained the same elements. Furthermore, the growth medium contained Fe, Co, and Eu which had been solubilized from the amphibolite. This indicates the presence, in this yeast strain, of active rockweathering abilities as well as of uptake mechanisms for solubilized rock components. PMID- 16345473 TI - Evidence for the existence of "survival factors" as an explanation for some peculiarities of yeast growth, especially in grape must of high sugar concentration. AB - The retardation and arrest of fermentation, observed before the complete sugar consumption of high-sugar grape must, come from an inhibition of the yeast metabolism during its decline phase and are variable with the strain. The addition of nutritional growth factors stimulates the initial growth of the yeast but is ineffective in the decline phase. Some substances, known previously as yeast anaerobic growth factors (sterols, oleanolic acid, oxytocin), in some conditions (initially aerated grape must and aerobically cultivated yeast) act by increasing the viability of the resting cells and prolonging their fermentation activity. These substances have been named "survival factors." PMID- 16345474 TI - Occurrence and Characterization of a Phosphoenolpyruvate: Glucose Phosphotransferase System in a Marine Bacterium, Serratia marinorubra. AB - The mechanism of d-glucose transport in the marine bacterium Serratia marinorubra was investigated. Uptake is mediated by a single, constitutive phosphoenolpyruvate:sugar phosphotransferase system (PTS), resulting in phosphorylation of d-glucose to d-glucose phosphate during transport. The system is saturable (K(m) = 6.4 x 10 M) and highly temperature dependent, with a Q(10) of 3.5 between 5 and 15 degrees C. The system is highly specific for d-glucose; structurally related sugars and sugar alcohols did not significantly compete with d-glucose for transport. The PTS requires Mg (K(m) = 2.5 x 10 M), but its activity is otherwise unaffected by salinity changes over the range tested (0 to 35 per thousand). S. marinorubra differs from other gram-negative organisms (Escherichia coli and Salmonella typhimurium) in that its glycerol (non-PTS substrate) permease is not regulated by the presence of glucose (PTS substrate). PMID- 16345475 TI - Capture of arginine at low concentrations by a marine psychrophilic bacterium. AB - The cells of the marine bacterium Ant-300 were found to take up arginine when this substrate was at low concentrations. The cells possessed an uptake system(s) that specifically transported l-arginine. The kinetic parameters for uptake appeared to differ when the cells were exposed to nanomolar and micromolar concentrations of the amino acid. Uptake over this concentration range functioned in the absence of an exogenous energy source, even after the cells had been preincubated in unsupplemented artificial seawater. Respiratory activity appeared to be a more important driving force for arginine uptake than adenosine 5' triphosphate hydrolysis. The cells also exhibited chemotaxis toward l-arginine. The minimum arginine concentration needed to elicit a chemotactic response was between 10 and 10 M. It is proposed that the capture of arginine by cells of Ant 300 in nutrient-depleted waters, which are typical of the open ocean, proceeds via high-affinity active transport, whereas in substrate-enriched seawater, capture involves chemotaxis and an active transport mechanism with reduced affinity for the substrate. PMID- 16345476 TI - Purification, Immobilization, and Some Properties of Glucose Isomerase from Streptomyces flavogriseus. AB - Glucose isomerase (EC 5.3.1.5) produced from Streptomyces flavogriseus was purified by fractionation with (NH(4))(2)SO(4) and chromatography on diethylaminoethyl (DEAE)-cellulose and DEAE-Sephadex A-50 columns. The purified enzyme was homogeneous as shown by ultracentrifugation and sodium dodecyl sulfate polyacrylamide gel electrophoresis. Benzyl DEAE-cellulose, triethylaminoethyl cellulose, and DEAE-cellulose were effective in the immobilization of partially purified glucose isomerase. Several differences in properties were found between purified soluble enzyme, immobilized enzyme (DEAE-cellulose-glucose isomerase), and heat-treated whole cells. Glucose and xylose served as substrate for the enzyme. Whole cells had the highest K(m) values for glucose and xylose; the soluble enzyme had the lowest values. The optimum temperature for activity of the soluble and immobilized enzymes was 70 degrees C; that for whole cells was 75 degrees C. The pH optimum for the three enzyme preparations was 7.5. Magnesium ion or Co was required for enzyme activity; an addition effect resulted from the presence of both Mg and Co. The enzyme activity was inhibited by Hg, Ag, or Cu. The conversion ratio of the enzyme for isomerization was about 50%. The soluble and immobilized enzymes showed a greater heat stability than whole cells. The soluble enzyme was stable over a slightly wider pH (5.0 to 9.0) range than the immobilized enzyme and whole cells (pH 5.5 to 9.0). The molecular weight of the enzyme determined by the sedimentation equilibrium method was 171,000. A tetrameric structure for the enzyme was also indicated. After operating at 70 degrees C for 5 days, the remaining enzyme activity of the immobilized enzyme and whole cells, which were used for the continuous isomerization of glucose in a plug-flow type of column in the presence of Mg and Co, was 75 and 55%, respectively. Elimination of Co decreased operational stability. PMID- 16345478 TI - Nitrate and nitrite reduction by microorganisms embedded in a filter paper incubated aerobically. AB - Pseudomonas aeruginosa, grown to steric saturation between the cellulose fibers of a filter paper, reduced nitrate or nitrite or both when the cell-filled paper was washed, transferred to phosphate buffer, nitrate, or nitrite or both, and glucose agar plates, and incubated under aerobiosis as resting cells. The biological nature of the reduction was ascertained by the use of nitrate and nitrite reductaseless mutants. The mesh of cellulose fibers was necessary to create a sufficient barrier to oxygen diffusion, since denitrification was not obtained within large and thick colonies of P. aeruginosa. When a soil suspension was used to inoculate the filter paper, ammonium and nitrite accumulated. Concomitant to nitrate reduction, the total nonvolatile inorganic nitrogen decreased and then increased as if part of it was immobilized to be subsequently mineralized. PMID- 16345477 TI - Influence of Environmental Factors on Antagonism of Fungi by Bacteria in Soil: Clay Minerals and pH. AB - The soil replica plating technique was used to evaluate the influence of clay minerals and pH on antagonistic interactions between fungi and bacteria in soil. In general, the antagonistic activity of bacteria towards filamentous fungi was greater in soil than on agar. The spread of Aspergillus niger through soil was inhibited by Serratia marcescens when the organisms were inoculated into separate sites in soil, and this antagonistic effect was maintained when the soil was amended with 3, 6, 9, or 12% (vol/vol) montmorillonite, whereas the addition of kaolinite at a concentration of 3% reduced the antagonism and at 6, 9, or 12% totally eliminated it. Similar results were obtained with the inhibition of A. niger by Agrobacterium radiobacter and of Penicillium vermiculatum by either S. marcescens or Nocardia paraffinae. When A. niger and S. marcescens were inoculated into the same soil site, A. niger was inhibited in all soils, regardless of clay content, although the extent of inhibition was greater as the concentration of montmorillonite, but not of kaolinite, increased. A. niger was inhibited more when inoculated as spores than as mycelial fragments and when inoculated 96 h after S. marcescens, but a 1% glucose solution reduced the amount of inhibition when the fungus was inoculated 96 h after the bacterium. When the pH of the soil-clay mixtures was altered, the amount of antagonism usually increased as the pH increased. Antagonism appeared to be related to the cation exchange capacity and the pH of the soil-clay mixtures. Bacillus cereus and another species of Bacillus showed no activity in soil towards A. niger under any of the environmental conditions tested, even though the Bacillus sp. significantly inhibited A. niger and seven other fungi on agar. PMID- 16345479 TI - Estimation of biodegradation potential of xenobiotic organic chemicals. AB - A method is described to estimate the biodegradation potential of soluble, insoluble, and unknown organic chemicals. The method consists of two stages: (i) generation of a microbial inoculum in a bench scale semicontinuous activated sludge system during which microorganisms are acclimated to test material and the removal of dissolved organic carbon is monitored and (ii) biodegradability testing (CO(2) evolution) in a defined minimal medium containing the test material as the sole carbon and energy source and a dilute bacterial inoculum obtained from the supernatant of homogenized activated sludge generated in the semicontinuous activated sludge system. Removal and biodegradation are measured using nonspecific methods, at initial concentrations of 5 to 10 mg of dissolved organic carbon per liter. Biodegradability data are accurately described by a nonlinear computer model which allows the rate and extent of biodegradation for different compounds to be compared and statistically examined. The evaluation of data generated in the combined removability-biodegradability system allows the biodegradation potential of a variety of xenobiotic organic chemicals to be estimated. PMID- 16345480 TI - Localization of active microorganisms in cheese by autoradiography. AB - By using an autoradiographic technique, one can follow, during ripening of a cheese, the distribution, size, and metabolic activity of microcolonies. Fragments of cheese were labeled with [H]leucine, fixed, and mounted in epoxy resin. After exposure and development, sections were examined by optical microscopy. In Camembert cheese, bacterial microcolonies synthesized protein rapidly during the beginning of the ripening process. At the end of the ripening process, active bacterial clusters were scarcer and of two types: (i) large microcolonies with reduced labeling, and (ii) microcolonies having the same size as those observed at the beginning of the ripening process, but with slight or no labeling. PMID- 16345481 TI - Nondividing, Bacteroid-Like Rhizobium trifolii: In Vitro Induction Via Nutrient Enrichment. AB - Rhizobium trifolii 0403 maintained in exponential phase via periodic dilution doubled in 210 min in mannitol-salts medium and doubled in 244 min in glycerolsalts. In both media, cell number and optical density increased in parallel. When exponentially growing cells in either medium were supplemented with a mixture of glucose, Casamino Acids, succinate, and yeast extract, optical density continued to increase but within less than the time required for one doubling, division ceased. The increase in optical density coupled with division cessation resulted in the formation of large, pleomorphic, nondividing cells. Large cells apparently increased in size as a result of swelling only at regions of most recent cell envelope synthesis. Greater than 95% of the cells in a population swelled, and commitment to swelling occurred within two doubling time equivalents. Swollen cells eventually reached a characteristic maximum size and exhibited osmotic fragility. PMID- 16345482 TI - Study of Developmental Stages of Methylosinus trichosporium with the Aid of Fluorescent-Antibody Staining Techniques. AB - When stained by using an indirect fluorescent-antibody technique, Methylosinus trichosporium displayed an uneven fluorescence. Exospores and the polar tips of some vegetative cells displayed a more intense fluorescence than the other cells. Cross-absorption of the specific anti-M. trichosporium immunoglobulin G with exospores resulted in no fluorescence of exospores or exospore regions of sporulating vegetative cells. This demonstrated that antigens were present on exospores and exospore regions of vegetative cells that are different from vegetative cell antigens. Taking advantage of this phenomenon, three fluorescentantibody staining techniques were developed which were used to study the life cycle of M. trichosporium. PMID- 16345483 TI - Hydrolytic enzyme production by Rhizobium. AB - Cellulase and hemicellulase activity was detected in temperate (infective and noninfective) and tropical strains (infective) of Rhizobium. Hydrolytic enzymes were initially detected by a cup-plate assay. The presence of cellulase and hemicellulase was confirmed by viscometric assay. Implications of the presence of these enzymes in Rhizobium are discussed. PMID- 16345484 TI - Characterization of azospirillum isolated from nitrogen-fixing roots of harvested sorghum plants. AB - Root segments of harvested sorghum plants had acetylene reduction activity ranging from 11 to 61 nmol of ethylene formed per h per g (dry weight). Five strains of Azospirillum brasilense sp. nov. were isolated from root segments. PMID- 16345485 TI - Mineralization capacity of bacteria and fungi from the rhizosphere-rhizoplane of a semiarid grassland. AB - A radiotracer glucose mineralization assay was used with streptomycin and actidione to monitor the relative seasonal contributions of bacteria and fungi to mineralization processes in soils derived from the rhizosphere-rhizoplane zone of plants from a shortgrass prairie ecosystem. Bacteria played a major role in glucose mineralization in both the rhizosphere and rhizoplane. These results indicate that the bacteria may play a greater role in glucose mineralization processes in the rhizosphere and rhizoplane zones of a semiarid grassland than would be assumed, based on available biomass estimates. This technique appears to be valuable for determining bacterial versus fungal contributions to glucose mineralization in the rhizosphere and rhizoplane and may be useful for measuring the decomposition of other more complex substances in this zone of intense microbial activity. PMID- 16345486 TI - Tenax-GC Extraction Technique for Residual Polychlorinated Biphenyl and Polyaromatic Hydrocarbon Analysis in Biodegradation Assays. AB - A rapid Tenax-GC extraction technique has been evaluated for use in conjunction with aqueous biodegradation assays for polyaromatic hydrocarbons and polychlorinated biphenyls. The method was quantitatively efficient and reproducible for phenanthrene, but variable and not quantitative for Aroclor 1254 (polychlorinated biphenyls). Aqueous sample volumes and varying concentrations of organic matter influenced polychlorinated biphenyl and polyaromatic hydrocarbon extraction efficiency. Phenanthrene recovery was decreased by soil extract but unaffected by spent bacteriological culture medium. Both types of organic matter caused significant reduction of Aroclor 1254 recovery. Polyaromatic hydrocarbon and polychlorinated biphenyl biodegradation assays, performed with reservoir samples, supported the laboratory evaluation. The study demonstrated the utility of the Tenax-GC extraction technique for phenanthrene analysis in biodegradation assessment; however, Tenax-GC extraction was not appropriate for Aroclor 1254 biodegradation studies. PMID- 16345487 TI - Phenanthrene biodegradation in freshwater environments. AB - Phenanthrene, a low-molecular-weight polycyclic aromatic hydrocarbon, was incubated with water samples from various reservoir systems in Tennessee to evaluate the potential for significant polycyclic aromatic hydrocarbon degradation by the indigenous microbial populations. Biodegradation was assessed by comparison of total polycyclic aromatic hydrocarbon substrate recovery in degradation flasks relative to sterile control flasks. During 1977 field studies, the mean phenanthrene biodegradation was approximately 80% after a 4-week incubation. Within a given habitat, 45% of the total variability in phenanthrene biodegradation was attributable to the physical, chemical, and microbiological site characteristics examined. Polycyclic aromatic hydrocarbon degradation was directly related to the historical environmental pollution of the sampling sites examined, the length of biodegradation assessment, temperature, and the molecular size of the polycyclic aromatic hydrocarbon substrate. PMID- 16345488 TI - Anaerobic methane oxidation: occurrence and ecology. AB - Anoxic sediments and digested sewage sludge anaerobically oxidized methane to carbon dioxide while producing methane. This strictly anaerobic process showed a temperature optimum between 25 and 37 degrees C, indicating an active microbial participation in this reaction. Methane oxidation in these anaerobic habitats was inhibited by oxygen. The rate of the oxidation followed the rate of methane production. The observed anoxic methane oxidation in Lake Mendota and digested sewage sludge was more sensitive to 2-bromoethanesulfonic acid than the simultaneous methane formation. Sulfate diminished methane formation as well as methane oxidation. However, in the presence of iron and sulfate the ratio of methane oxidized to methane formed increased markedly. Manganese dioxide and higher partial pressures of methane also stimulated the oxidation. The rate of methane oxidation in untreated samples was approximately 2% of the CH(4) production rate in Lake Mendota sediments and 8% of that in digested sludge. This percentage could be increased up to 90% in sludge in the presence of 10 mM ferrous sulfate and at a partial pressure of methane of 20 atm (2,027 kPa). PMID- 16345489 TI - Mechanism of Ozone Inactivation of Bacteriophage f2. AB - The inactivation kinetics of bacteriophage f2 were studied by using ozone under controlled laboratory conditions. The phage were rapidly inactivated during the first 5 s of the reaction by 5 and 7 logs at ozone concentrations of 0.09 and 0.8 mg/liter, respectively. During the next 10 min, the phage were further inactivated at a slower rate in both treatments. The [H]uridine-labeled f2 phage and its ribonucleic acid (RNA) were examined to elucidate the mechanism of ozone inactivation, utilizing adsorption to host bacteria, sucrose density gradient analysis, and electron microscopy. The specific adsorption of the phage was reduced by ozonation in the same pattern as plaque-forming unit reduction. RNA was released from the phage particles during ozonation, although it had reduced infectivity for spheroplasts. Electron microscopic examination showed that the phage coat was broken by ozonation into many protein subunit pieces and that the specific adsorption of the phage to host pili was inversely related to the extent of phage breakage. The RNA enclosed in the phage coat was inactivated less by ozonation than were whole phage, but inactivated more than naked RNA. These findings suggest that ozone breaks the protein capsid into subunits, liberating RNA and disrupting adsorption to the host pili, and that the RNA may be secondarily sheared by a reduction with and/or without the coat protein molecules, which have been modified by ozonation. PMID- 16345490 TI - Association of azospirillum with grass roots. AB - The association between grass roots and Azospirillum brasilense Sp 7 was investigated by the Fahraeus slide technique, using nitrogen-free medium. Young inoculated roots of pearl millet and guinea grass produced more mucilaginous sheath (mucigel), root hairs, and lateral roots than did uninoculated sterile controls. The bacteria were found within the mucigel that accumulated on the root cap and along the root axes. Adherent bacteria were associated with granular material on root hairs and fibrillar material on undifferentiated epidermal cells. Significantly fewer numbers of azospirilla attached to millet root hairs when the roots were grown in culture medium supplemented with 5 mM potassium nitrate. Under these growth conditions, bacterial attachment to undifferentiated epidermal cells was unaffected. Aseptically collected root exudate from pearl millet contained substances which bound to azospirilla and promoted their adsorption to the root hairs. This activity was associated with nondialyzable and proteasesensitive substances in root exudate. Millet root hairs adsorbed azospirilla in significantly higher numbers than cells of Rhizobium, Pseudomonas, Azotobacter, Klebsiella, or Escherichia. Pectolytic activities, including pectin transeliminase and endopolygalacturonase, were detected in pure cultures of A. brasilense when this species was grown in a medium containing pectin. These studies describe colonization of grass root surfaces by A. brasilense and provide a possible explanation for the limited colonization of intercellular spaces of the outer root cortex. PMID- 16345491 TI - Silicates as nonspecific adsorbents of bacteriophage: a model for purification of water from viruses. AB - Amorphous silicates having small particles with a large surface area were found to have high nonspecific adsorption capacity. Investigations with Escherichia coli T4 bacteriophage have shown adsorption capacity of up to 99.9% on different tested silicates. Optimal conditions for the adsorption of T4 bacteriophage on various commercially available silicates were determined. A continuous-flow adsorption of T4 bacteriophage on silicates appears to be a model system for the removal of viruses from potable water and sewage. PMID- 16345492 TI - Indicator technique for antimetabolic toxin production by phytopathogenic species of pseudomonas. AB - A simple bacteriological technique involving inhibition zone production on a lawn of Escherichia coli was developed to detect antimetabolite toxin production by phytopathogenic species of Pseudomonas. It was established that the mechanism of E. coli inhibition paralleled that of phytotoxin-induced chlorosis of plant tissue. Derivatives of Pseudomonas tabaci and Pseudomonas phaseolicola which did not produce antimetabolite were readily identified by use of this technique. The presence of plasmid DNA in P. tabaci strains was demonstrated, but no physical evidence for plasmid involvement in tabtoxin production was found. The role of antimetabolite production in the pathogenicity of P. phaseolicola was also investigated. The method was extended to show that strains of P. maculicola, P. syringae, and P. coronofaciens also produce antimetabolites. PMID- 16345493 TI - A thick-walled organism isolated from the cockroach gut by using a spent medium technique. AB - By using a conditioning technique whereby complex media are inoculated several times with bacteria from the hindgut of the cockroach Eublaberus posticus, a succession of bacterial types occurred. An obligately anaerobic, pleomorphic, thick-walled, gram-positive organism is described which was isolated by this culturing technique. PMID- 16345494 TI - Direct Enzymatic Assay for Alcohol Oxidase, Alcohol Dehydrogenase, and Formaldehyde Dehydrogenase in Colonies of Hansenula polymorpha. AB - A procedure is described for the qualitative direct identification of alcohol oxidase, alcohol dehydrogenase, and formaldehyde dehydrogenase in yeast colonies. The method has been applied successfully to isolate mutants of Hansenula polymorpha with altered glucose repression of alcohol oxidase. PMID- 16345495 TI - Fermentation Methods for Protein Enrichment of Cassava and Corn with Candida tropicalis. AB - Candida tropicalis grows on soluble starch, corn, and cassava powders without requiring that these substrates be previously hydrolyzed. C. tropicalis possesses the enzyme needed to hydrolyze starch, namely, an alpha-amylase. That property has been used to develop a fermentation process whereby C. tropicalis can be grown directly on corn or cassava powders so that the resultant mixture of biomass and residual corn or cassava contains about 20% protein, which represents a balanced diet for either animal fodder or human food. The fact that no extra enzymes are required to hydrolyze starch results in a particularly efficient way of improving the nutritional value of amylaceous products, through a single-step fermentation process. PMID- 16345496 TI - Critical Reactions in Fluorobenzoic Acid Degradation by Pseudomonas sp. B13. AB - 3-Chlorobenzoate-grown cells of Pseudomonas sp. B13 readily cometabolized monofluorobenzoates. A catabolic pathway for the isomeric fluorobenzoates is proposed on the basis of key metabolites isolated. Only 4-fluorobenzoate was utilized and totally degraded after a short period of adaptation. The isoenzymes for total degradation of chlorocatechols, being found during growth with 3 chlorobenzoate or 4-chlorophenol, were not induced in the presence of fluorobenzoates. Correspondingly, only the ordinary enzymes of the benzoate pathway were detected in 4-fluorobenzoate-grown cells. Ring cleavage of 3 fluorocatechol was recognized as a critical step in 3-fluorobenzoate degradation. 2-Fluoro-cis,cis-muconic acid was identified as a dead-end metabolite from 2- and 3-fluorobenzoate catabolism. During 2-fluorobenzoate cometabolism, fluoride is eliminated by the initial dioxygenation. PMID- 16345497 TI - Adaptation of Alcaligenes eutrophus B9 and Pseudomonas sp. B13 to 2 Fluorobenzoate as Growth Substrate. AB - Alcaligenes eutrophus B9 and Pseudomonas sp. B13 could be adapted to 2 fluorobenzoate as the sole source of carbon and energy. The ability of the A. eutrophus B9 to use this new substrate is clearly based on the defective dihydrodihydroxybenzoate dehydrogenase. Nontoxic 6-fluoro-3,5-cyclohexadiene-1,2 diol-1-carboxylic acid is accumulated instead of 3-fluorocatechol. About 84% of the substrate is dioxygenated to catechol and utilized via the 3-oxoadipate pathway. During continuous adaptation of Pseudomonas sp. B13 regioselectivity of dioxygenation of 2-fluorobenzoate is drastically changed in favor of a 1,2 attack. Consequently, approximately 97% of the substrate is utilized via catechol. A three- to fourfold overproduction of key enzymes of the 3-oxoadipate pathway compensates for the slower turnover rates of the fluorinated substrates. PMID- 16345498 TI - Kinetics of manganese oxidation by cell-free extracts of bacteria isolated from manganese concretions from soil. AB - A study of the kinetics of Mn oxidation catalyzed by cell extracts of two bacterial isolates (E(1), Pseudomonas III [new isolate] and E(4), Citrobacter freundii) isolated from the core of manganese concretions from Greek soils is presented. The reaction velocity of Mn oxidation was determined from the rate of consumption of Mn. The oxidation of Mn was followed by measuring changes in Mn concentration by activation analysis and by atomic absorption spectrophotometry. The reaction velocity was directly proportional to cell extract concentration when the reaction time was 1 h. At longer reaction times, the relationship deviated from linearity because substrate concentration became limiting. The rate of Mn oxidation increased with the Mn concentration. Analysis of the results by application of the integrated Michaelis equation for determining Michaelis constants and maximal velocities either in the presence (K(m) = 3.33 mumol/ml and V(max) = 1.25 mumol/ml.h) or in the absence of maleate buffer (K(m) = 2.52 mumol/ml and V(max) = 2.04 mumol/ml.h) indicated a strong affinity between the oxidizing system and manganese. All results in this study are consistent with an enzymatic manganese-oxidizing system and give an indication of the mechanism of biological Mn oxidation in soil which differs from that in the marine environment. PMID- 16345499 TI - Production and biological activity of secalonic Acid d. AB - Twenty isolates of Penicillium oxalicum produced secalonic acid as their major secondary metabolite. Fermentation conditions were determined for toxin production in grain and liquid media. The 50% lethal dose value for mice ranged from 26.5 to 51.7 mg/kg dependent on animal strain and sex, males being more susceptible than females. Secalonic acid was nontoxic and nonteratogenic to the chicken embryo and exhibited poor antibiotic properties. Its potential role in mycotoxicoses is discussed. PMID- 16345500 TI - Metabolism of Dibenzo-p-Dioxin and Chlorinated Dibenzo-p- Dioxins by a Beijerinckia Species. AB - Whole cells of the parent strain of Beijerinckia, grown with succinate and biphenyl, oxidized dibenzo-p-dioxin and several chlorinated dioxins. The rate of oxidation of the chlorinated dibenzo-p-dioxins decreased with an increasing degree of chlorine substitution. A mutant strain (B8/36) of Beijerinckia oxidized dibenzo-p-dioxin to cis-1,2-dihydroxy-1,2-dihydrodibenzo-p-dioxin. The mutant organism also oxidized two monochlorinated dibenzo-p-dioxins to cis-dihydrodiols. No metabolites were detected from two dichlorinated dibenzo-p-dioxins. Growth of the parent strain of Beijerinckia on succinate was inhibited after 4 h when 0.05% dibenzo-p-dioxin was present in the culture medium. Resting cell suspensions of the parent organism, previously grown with succinate and biphenyl, oxidized dibenzo-p-dioxin to a compound identified as 1,2-dihydroxydibenzo-p-dioxin. Further degradation of this metabolite was not detected, as the compound was found to be a potent mixed-type inhibitor of two ring-fission oxygenases present in this organism. PMID- 16345501 TI - Promotion of infection thread formation by substances from Rhizobium. AB - An extrinsic substance (ES-6000) was isolated from the periplasmic space of Rhizobium trifolii (strain 4S) cells by osmotic shock, using a high-density sucrose solution. This substance promoted infection thread formation in root hairs of white clover when inoculated together with the infectious strain (4S). However, ES-6000 obtained from another rhizobial species and from strain A1, which is a noninfectious mutant strain obtained from strain 4S, did not have this effect. The promoter in the ES-6000 from strain 4S is a relatively small molecule since it passed through a hollow-fiber membrane (molecular weight, 6,000). This substance was also recognized as an R(f) 0.1 fraction by paper chromatography. Sucrose was effective in promoting nodulation and root elongation. PMID- 16345502 TI - Planktonic marine luminous bacteria: species distribution in the water column. AB - Luminous bacteria were isolated from oceanic water samples taken throughout the upper 1,000 m and ranged in density from 0.4 to 30 colony-forming units per 100 ml. Generally, two peaks in abundance were detected: one in the upper 100 m of the water column, which consisted primarily of Beneckea spp.; and a second between 250 and 1,000 m, which consisted almost entirely of Photobacterium phosphoreum. The population of P. phosphoreum remained relatively stable in abundance at one station that was visited three times over a period of 6 months. However, the abundance of luminous Beneckea spp. isolated from the upper waters fluctuated considerably; they were, as high as 30 colony-forming units per 100 ml in the spring and were not detected in the winter. Water samples from depths of 4,000 to 7,000 m contained less than 0.1 luminous colony-forming unit per 100 ml. The apparent vertical stratification of two taxa of oceanic luminous bacteria may reflect not only differences in physiology, but also depth-related, species specific symbiotic associations. PMID- 16345503 TI - Effect of carbon dioxide on growth of meat spoilage bacteria. AB - The ability of CO(2) to inhibit respiration and growth of representative strains of seven species of meat spoilage bacteria was examined. Enterobacter and Microbacterium thermosphactum were unaffected by CO(2). Both respiration and growth of the other species were inhibited. With four of the species (fluorescent and nonfluorescent Pseudomonas, Alteromonas putrefaciens, and Yersinia enterocolitica), the inhibition pattern in a complex medium was similar, and inhibition was incomplete and reached a maximum level at comparatively low concentrations of CO(2). With Acinetobacter, inhibition continued to increase with increasing CO(2) concentration. The degree of inhibition with a constant concentration of CO(2) in solution increased with decreasing temperature for all CO(2)-susceptible species except the nonfluorescent Pseudomonas. Anaerobic growth of CO(2)-susceptible facultative anaerobes was unaffected by CO(2). PMID- 16345504 TI - Biological Process for Converting Naphthalene to cis-1,2-Dihydroxy-1,2 Dihydronaphthalene. AB - A biological process for converting naphthalene to cis-1,2-dihydroxy-1,2 dihydronaphthalene (DHD) catalyzed by Pseudomonas putida strain 119 was optimized in flask experiments. These studies revealed the following: (i) P. putida 119 can propagate efficiently and produce DHD when supplied one of several carbon sources and naphthalene; (ii) maximum DHD production by P. putida 119 occurs in logarithmic-growth-phase cells and decreases at various rates in the stationary growth phase, depending upon the carbon source used; (iii) several analogs of salicylic acid can be used as effective inducers of naphthalene metabolism in P. putida cells growing on glucose; and (iv) the addition of chemical surfactants to naphthalene-cell (P. putida 119) mixtures stimulates DHD production. PMID- 16345505 TI - Enzymatic Synthesis of l-Carnitine by Reduction of an Achiral Precursor: the Problem of Reduced Nicotinamide Adenine Dinucleotide Recycling. AB - Synthesis of l-carnitine has been carried out by the enzymatic reduction of the carbonyl group of the achiral precursor 3-dehydrocarnitine with the oxidized nicotinamide adenine dinucleotide-linked carnitine dehydrogenase. Various enzymatic or chemical systems have been tested to regenerate the reduced nicotinamide adenine dinucleotide oxidized in the reduction of 3 dehydrocarnitine. Because of the instability of this compound in aqueous solutions, it was added by continuous feeding as a rate-limiting constituent in the reaction mixture. Under these conditions, conversion yields of 95% were achieved with the glucose plus glucose dehydrogenase system. A total number of 530 reduced nicotinamide adenine dinucleotide recyclings was obtained with this system for a production of 45 g of l-carnitine per liter. The stabilities of the oxidized nicotinamide adenine dinucleotide and the reduced nicotinamide adenine dinucleotide have been determined at various pH values. In view of these results, several possible strategies for enzymatic syntheses with the reduced nicotinamide adenine dinucleotide as a regenerable coenzyme are discussed. PMID- 16345506 TI - Water-to-Air Fractionation of Bacteria. AB - Differences in the ability of bacterial species to be transported from water to air by bursting bubbles were investigated. Bubbles were generated in suspensions of mixed bacterial cultures, and the concentration was measured for each species in the top jet drop ejected. This concentration divided by the concentration in the bulk menstruum is the concentration factor (CF). Bubbles were generated 2 cm below the liquid surface, and jet drops with diameters from 34 to 136 mum were studied. Serratia marinorubra and Micrococcus euryhalis had CFs which were generally 10 to 100 times greater than those of Escherichia coli, Pseudomonas bathycetes, and spores of Bacillus subtilis. P. bathycetes never had a CF significantly greater than 1 at any drop size, and spores of B. subtilis had a maximum CF of 4. E. coli had a maximum CF of 6 for 1- or 2-day old cultures, but this increased to 80 when a 5-day-old culture was used. This change in the CF with age of the cells indicates that composition of the cell may be a factor influencing its ability to concentrate in jet drops. PMID- 16345507 TI - Nitrogen fixation by photosynthetic bacteria in lowland rice culture. AB - Propanil (3',4'-dichloropropionanilide) was a potent inhibitor of the nitrogenase activity of blue-green algae (cyanobacteria) in flooded soil, but the herbicide at comparable concentrations was not toxic to rice, protozoa, and nitrogen-fixing bacteria. Ethanol-amended flooded soils treated with propanil exhibited higher rates of nitrogenase activity than those not treated with the herbicide. The enhanced nitrogenase activity in propanil-treated soils was associated with a rise in the population of purple sulfur bacteria, especially of cells resembling Chromatium and Thiospirillum. By employing propanil and a means of excluding light from the floodwater to prevent the development of phototrophs during rice growth under lowland conditions, the relative activities of blue-green algae, photosynthetic bacteria, and the rhizosphere microflora were determined. The results suggest that the potential contribution of photosynthetic bacteria may be quite high. PMID- 16345508 TI - Laboratory comparison of the effectiveness of several algicides on isolated swimming pool algae. AB - The most frequently encountered species of algae found in swimming pools in the Phoenix metropolitan area were used to evaluate the laboratory effectiveness of five commercially available pool chemicals used for algal control. The pool algae used were the xanthophyte Pleurochloris pyrenoidosa, the chlorophyte Oocystis sp., and the cyanophytes Phormidium minnesotense and Plectonema sp. Pad Algae Kill (a chlorine derivative) was effective in the control of all test organisms. Algaedyn, a silver-containing algicide, was effective on P. minnesotense and Plectonema sp., but caused only a slight inhibition in the growth of P. pyrenoidosa and Oocystis sp. Quarternary ammonium (Padicide) was more effective in controlling the growth of Phormidium and Plectonema than Pleurochloris and Oocystis. Algimycin (herbicide) only reduced the growth of Oocystis and was ineffective on the other species. Bio-Gard (copper) reduced the growth of Pleurochloris, but had no effect on the other test organisms. The technique used to quantify the influence of algicides on isolated pool algae appears to be adaptable to those algae that form distinct colonies on an agar substratum. PMID- 16345509 TI - Influence of environmental factors on antagonism of fungi by bacteria in soil: nutrient levels. AB - The addition of 0.25, 0.5, or 1.0% glucose to a soil (K) amended with either 6% kaolinite (K6K) or montmorillonite (K6M) or the adjustment of the C/N ratio of the soils from 23/1 to 10/1 with NH(4)NO(3) eliminated the inhibition of Aspergillus niger by Serratia marcescens, regardless of whether the fungus and bacterium were inoculated into the same or separate sites in the soils. The adjustment of the C/N ratio to 15/1 or of the C/P ratio from 1,000/1 to 100/1 with KH(2)PO(4) did not eliminate the antagonism. However, with the higher glucose and NH(4)NO(3) amendments, S. marcescens died out in the K and K6K (but not in the K6M) soils, apparently due to reductions in pH that resulted from the increased metabolism induced by added nutrients. In soils amended with CaCO(3), S. marcescens did not die out, but the inhibition of A. niger by S. marcescens or Agrobacterium radiobacter was eliminated or reduced by the addition of glucose, but not of NH(4)NO(3), and was influenced by the clay mineralogy and pH of the soils. When NH(4)NO(3) was added to the soils adjusted with CaCO(3) to pH values above 6.0, growth of A. niger was inhibited, regardless of whether bacteria were present or not, as a result of the volatilization of NH(3). Bacillus cereus and another species of Bacillus did not inhibit A. niger under any of the environmental conditions. There was a direct correlation between the degree of inhibition and the rate of glucose utilization by the various bacteria, indicating that the antagonism of A. niger by some bacteria in soil was the result primarily of a competition for carbon and that this competition was influenced by other environmental factors, such as pH and clay mineralogy. PMID- 16345510 TI - Destruction by anaerobic mesophilic and thermophilic digestion of viruses and indicator bacteria indigenous to domestic sludges. AB - In raw sludges and in mesophilically and thermophilically digested anaerobic sludges, large variations in numbers of viruses occurred over narrow ranges of numbers of fecal coliforms, total coliforms, and fecal streptococci, demonstrating that the bacteria were poor quantitative reflectors of the numbers of the viruses detected. Mesophilic and thermophilic digestion of anaerobic sludges destroyed all three indicator bacteria more rapidly than such digestion destroyed the viruses. The relative rates for the destruction of viruses, fecal coliforms, and fecal streptococci in the digested sludges were consistent over the 17-month study. Fecal coliforms were 7 to 8 times more sensitive than the viruses to mesophilic digestion and 9 to 10 times more sensitive to thermophilic digestion. Total coliforms were even more sensitive. The rates at which fecal streptococci were destroyed by mesophilic and thermophilic digestion of anaerobic sludges approached those at which the viruses were destroyed by those processes; this suggested that the rates at which fecal streptococci in sludges are destroyed by those processes may serve as useful indicators for the rates at which viruses in sludges are destroyed by those processes. PMID- 16345511 TI - Use of a cellulase-derepressed mutant of cellulomonas in the production of a single-cell protein product from cellulose. AB - A cellulase-derepressed mutant of a Cellulomonas species was used to produce single-cell protein from crystalline cellulose. In preliminary tests, maximum yield of single-cell protein was obtained at 30 degrees C (pH 7.0) with urea as the nitrogen source. A continuous-flow foam flotation procedure was developed for rapid and efficient separation of bacteria from the culture liquid and cellulose residue. A pH of 4.5 was optimum for foam flotation of this organism. In preliminary trials, recovery was 85% of the cells with the flotation procedure. Cellulomonas was 68% true protein and had an essential amino acid profile featuring a high lysine content (6.5% of protein). The Cellulomonas product was evaluated nutritionally with weanling rats. The net protein utilization value for the protein supplemented with methionine was 50.4% Weight gain of rats on the Cellulomonas diet was similar to that of rats fed a casein diet. PMID- 16345512 TI - Kinetics of Growth and Amylase Production of Saccharomycopsis fibuligera on Potato Processing Wastewater. AB - The kinetics of growth and amylase production of Saccharomycopsis fibuligera were studied in a chemostat on a synthetic potato processing blancher water. Dilution rates (D) from 0.101 to 0.480 h were examined. A mathematical model based on the Monod equation was developed. The yield of cell mass from carbohydrates was constant and equal to 0.84. The maximum specific growth rate and the Monod constant were determined to be 0.596 h and 0.226 mg/ml, respectively. An equation for the steady-state starch concentrations was empirically derived. The steady state noncarbohydrate carbon levels rose linearly with D. Reducing sugars were the growth-limiting substrate, and their steady-state levels conformed to Monod kinetics. The yield of amylase from the cell mass (Y(z)) declined as D rose and was described by the equation Y(z) = (-8.005D + 4.076). The model predicted that the maximum production of cell mass should occur at D = 0.35 h and the maximum production of amylase should occur at D = 0.22 h. The mathematical model presented agreed with the experimental results in its prediction of the steady state level of reducing sugar, starch, cell mass, and amylase concentrations as well as the productivity of amylase. PMID- 16345513 TI - Growth of fish cell lines on microcarriers. AB - Microcarrier beads were evaluated as substrates for the propagation of five anchorage-dependent fish cell lines. Growth of rainbow trout gonad (RTG-2) and Atlantic salmon cells was limited on microcarriers maintained in suspension. However, stationary microcarriers were suitable substrates for the growth of RTG 2, AS, Chinook salmon embryo (CHSE-214), and fathead minnow cells. Cell yields ranged from 2 x 10 to 2.9 x 10 cells per ml, representing 7- to 10-fold increases over the initial cell concentrations. The yield of new RTG-2 cells per unit volume of growth medium was 2.8 times greater in microcarrier cultures than in standard monolayer cultures. Northern pike cells failed to grow on microcarriers. Yields of infectious pancreatic necrosis virus propagated in microcarrier cultures of RTG-2 cells were more than twice the yields in standard monolayer cultures. The greater economy of microcarrier cultures in terms of growth vessel and medium requirements holds great promise for the large-scale production of anchorage-dependent fish cell cultures and fish viruses. PMID- 16345514 TI - Use of two-dimensional polyacrylamide gel electrophoresis to identify and classify Rhizobium strains. AB - Fifty-seven strains of various Rhizobium species were analyzed by two-dimensional gel electrophoresis. Since the protein pattern on such gels is a reflection of the genetic background of the tested strains, similarities in pattern allowed us to estimate the relatedness between these strains. All group II rhizobia (slow growing) were closely related and were very distinct from group I rhizobia (fast growing). Rhizobium meliloti strains formed a distinct group. The collection of R. leguminosarum and R. trifolii strains together formed another distinct group. Although there were some similarities within the R. phaseoli, sesbania rhizobia, and lotus rhizobia, the members within these seemed much more diverse than the members of the above groups. The technique also is useful to determine whether two unknown strains are identical. PMID- 16345515 TI - Rapid membrane filtration-epifluorescent microscopy technique for direct enumeration of bacteria in raw milk. AB - Membrane filtration and epifluorescent microscopy were used for the direct enumeration of bacteria in raw milk. Somatic cells were lysed by treatment with trypsin and Triton X-100 so that 2 ml of milk containing up to 5 x 10 somatic cells/ml could be filtered. The majority of the bacteria (ca. 80%) remained intact and were concentrated on the membrane. After being stained with acridine organe, the bacteria fluoresced under ultraviolet light and could easily be counted. The clump count of orange fluorescing cells on the membrane correlated well (r = 0.91) with the corresponding plate count for farm, tanker, and silo milks. Differences between counts obtained by different operators and between the membrane clump count and plate count were not significant. The technique is rapid, taking less than 25 min, inexpensive, costing less than 50 cents per sample, and is suitable for milks containing 5 x 10 to 5 x 10 bacteria per ml. PMID- 16345516 TI - Isolation and analysis of molds from soy sauce koji in Thailand. AB - Five different isolates of Aspergillus and one of Mucor were compared with a Japanese commercial strain of Aspergillus oryzae for proteolytic activity on wheat bran substrate. One isolate of Aspergillus with superior protease production, identified as Aspergillus flavus var. columnaris, showed no detectable aflatoxin production on glutinous rice or soybean substrate. Preliminary tests using this fungus as a koji mold in a traditionally operated factory resulted in a soy sauce superior in quality to that usually produced. PMID- 16345517 TI - Methanogenic decomposition of ferulic Acid, a model lignin derivative. AB - Ferulic acid, a model lignin derivative, was observed to be biodegradable to methane and carbon dioxide under strict anaerobic conditions. This conversion appears to be carried out by a consortium of bacteria similar to that previously described for the methanogenic degradation of benzoic acid. A temporary buildup of acetate in these cultures indicates that it is a likely intermediate and precursor for methane formation. An analog of coenzyme M, 2-bromoethanesulfonic acid (BESA), inhibited gas production and enhanced the buildup of propionate, butyrate, isobutyrate, and isovalerate. Phenylacetate, cinnamate, 3 phenylpropionate, benzoate, cyclohexane carboxylate, adipate, and pimelate were also detected in BESA-inhibited cultures. A pathway is proposed which includes these various acids as possible intermediates in the methanogenic degradation of ferulic acid. This model overlaps previously described benzoic acid degradation pathways, suggesting that this type of anaerobic degradation may be common for aromatic compounds. PMID- 16345518 TI - Interaction Between a Non-Nodulating and an Ineffective Mutant of Rhizobium trifolli Resulting in Effective (Nitrogen-Fixing) Nodulation. AB - A noninvasive (non-nodulating) mutant of Rhizobium trifolii when mixed with an ineffective (non-nitrogen-fixing) mutant gives rise to effective (nitrogen fixing) nodules. PMID- 16345519 TI - Sulfide Production from Cysteine by Desulfovibrio desulfuricans. AB - Two rumen nitrate-reducing isolates of Desulfovibrio desulfuricans were found to hydrolyze cysteine with the production of sulfide and pyruvate. When cultured on agar medium containing yeast extract with nitrate as the primary electron acceptor and ferrous chloride as the indicator, blackening of colonies occurred. The blackening of colonies appeared sooner and was more intense when either cysteine or sulfate was added to the culture medium with nitrate present. PMID- 16345520 TI - Use of Two-Dimensional Polyacrylamide Electrophoresis to Demonstrate that Putative Rhizobium Cross-Inoculation Mutants Actually Are Contaminants. AB - Two-dimensional polyacrylamide electrophoresis was used to determine that mutants of Rhizobium trifolii DT6, claimed to be capable of effectively nodulating soybeans, were actually Rhizobium japonicum 110 contaminants isolated from the parent DT6 culture. PMID- 16345521 TI - Desulfuration of dialkyl thiophosphoric acids by a pseudomonad. AB - A strain of Pseudomonas acidovorans used the organophosphorus pesticide breakdown products, ionic O,O-diethyl phosphorothioate and ionic O,O-diethyl phosphorodithioate, as sulfur sources. The growth yields from the thiophosphates and sulfate were 3.6 to 4.1 kg of protein per mol of sulfur. Elemental sulfur and sulfide also served as sulfur sources but gave lower growth yields. PMID- 16345522 TI - Density Centrifugation Method for Recovering Rhizobium spp. from Soil for Fluorescent-Antibody Studies. AB - A density centrifugation procedure has been developed as a replacement for soil flocculation and clarification steps employed in quantitative fluorescent antibody studies on Rhizobium in soils. Near-quantitative recovery of added cells of two strains of Rhizobium japonicum and two strains of R. phaseoli was achieved from six soils with various properties. It is proposed that this technique may prove useful in separating other soil microorganisms from soil particles in ecological studies employing fluorescent-antibody techniques. PMID- 16345523 TI - Assimilation of lemonade-processing wastewater by yeasts. AB - Of the five species of yeasts examined in shake-flask studies at 30 degrees C, only Candida utilis and Saccharomyces fragilis gave higher cell yields (>2.5 g /liter) in 32 h and reduced approximately 87% of the biological oxygen demand in lemonade-processing wastewater fortified with 0.05% ammonium sulfate and 0.01% potassium phosphate. The latter, however, settled more rapidly than the former. PMID- 16345524 TI - Bacterial Flora of the Schistosome Vector Snail Biomphalaria glabrata. AB - [This corrects the article on p. 671 in vol. 38.]. PMID- 16345525 TI - Specific inhibition of nitrite oxidation by chlorate and its use in assessing nitrification in soils and sediments. AB - A method was developed to determine the ammonium oxidation rate (potential) of unenriched natural samples by measuring the nitrite produced in shaken slurries. Addition of chlorate to the samples prevented nitrite from being oxidized to nitrate. The effectiveness and specificity of chlorate were tested with pure cultures of nitrite and ammonium oxidizers, as well as in soil and sediment slurries. It was concluded that chlorate had relatively little inhibitory effect on ammonium oxidation. However, under some conditions chlorate was not completely effective in blocking nitrite oxidation, and the causes of this were investigated. The technique was designed to check for incomplete blockage. PMID- 16345526 TI - Cell surface measurements in hydrocarbon and carbohydrate fermentations. AB - Acinetobacter calcoaceticus was grown in 11-liter batch fermentations with hexadecane or sodium citrate as the sole source of carbon. Surface and interfacial tension measurements of the microbial broth indicated that surface active compounds were being produced only during growth on the hydrocarbon substrate. Contact angle measurements of an aqueous drop on a smooth lawn of cells in a hexadecane bath indicated a highly hydrophobic surface of the cells in the initial stages of the hydrocarbon fermentation (120 degrees contact angle). At this stage, the entire cell population was bound to the hydrocarbon-aqueous interface. The contact angle dropped rapidly to approximately 45 degrees after 14 h into the fermentation. This coincided with a shift of the cell population to the aqueous phase. Thus, the cells demonstrated more hydrophilic characteristics in the later stages of the fermentation. Contact angles on cells grown on sodium citrate ranged from 18 to 24 degrees throughout the fermentation. The cells appear to be highly hydrophilic during growth on a soluble substrate. From the contact angle and aqueous-hydrocarbon interfacial tension, the surface free energy of the cells was calculated along with the cell-aqueous and cell hydrocarbon interfacial tension. The results of these measurements were useful in quantitatively evaluating the hydrophobic nature of the cell surface during growth on hydrocarbons and comparing it with the hydrophilic nature of the cell surface during growth on a soluble substrate. PMID- 16345527 TI - Metabolism of Radiolabeled beta-Guaiacyl Ether-Linked Lignin Dimeric Compounds by Phanerochaete chrysosporium. AB - Phanerochaete chrysosporium metabolized the radiolabeled lignin model compounds [gamma-C]guaiacylglycerol-beta-guaiacyl ether and [4-methoxy-C]veratrylglycerol beta-guaiacyl ether (VI) to CO(2) in stationary and in shaking cultures. CO(2) evolution was greater in stationary culture. CO(2) evolution from [gamma C]guaiacyl-glycerol-beta-guaiacyl ether and [4-methoxy-C]veratrylglycerol-beta guaiacyl ether in stationary cultures was two- to threefold greater when 100% O(2) rather than air (21% O(2)) was the gas phase above the cultures. CO(2) evolution from the metabolism of the substrates occurred only as the culture entered the stationary phase of growth. The presence of substrate levels of nitrogen in the medium suppressed CO(2) evolution from both substrates in stationary cultures. [C]veratryl alcohol and 4-ethoxy-3-methoxybenzyl alcohol were formed as products of the metabolism of VI and 4-ethoxy-3 methoxyphenylglycerol-beta-guaiacyl ether, respectively. PMID- 16345528 TI - Fixation of dinitrogen-15 associated with rice plants. AB - Rice plants (IR26 and Latisail) obtained at near heading stage from a wetland field were transferred to water culture and exposed to N(2) in a gas-tight growth chamber for 7 days to measure N(2)-fixing activities associated with the rice. The activities measured varied from 6.5 to 11.6 mumol of N(2) fixed per hill per day. The outer leaf sheath had about 2.5 times higher N(2)-fixing activities per unit weight than the root. Slight activities were also found in the basal node and inner leaf sheath. Wrapping basal parts of the stem with aluminum foil did not decrease the activities of N(2) fixation in these parts. Thus, the outer leaf sheath as well as the root are N(2)-fixing sites in rice plants. N(2) fixation found in above-ground parts is not due to photoautotrophic organisms. Less than 10% of the fixed nitrogen was translocated from the fixing sites to the leaf blades and the young panicles. PMID- 16345529 TI - Sediment distribution of methanogenic bacteria in lake erie and cleveland harbor. AB - The direct fluorescent-antibody technique was employed to determine the distribution patterns of four species of methanogens in the sediments of Lake Erie and Cleveland Harbor. Methanobacterium ruminantium was the most numerous methanogen found in regions of high-organic-silt sediments. The population of this species ranged from 10 to 10 cells/g of dry sediment. Methanobacterium strain MoH and Methanosarcina barkeri were identified in sand-silt, clay, or sand sediments. These methanogens ranged in density from 10 to 10 cells/g of dry sediment. Methanospirillum hungatii was observed only after an organic enrichment was performed on Cleveland Harbor sediments. The seasonal and selective sediment distribution of these methanogens appears to be related to the type and concentration of carbon as substrate as well as to the activities of heterotrophic and sulfate-reducing bacteria. PMID- 16345530 TI - Growth of iron-oxidizing thiobacilli in the presence of chalcopyrite and galena. AB - Iron-oxidizing thiobacilli were adapted to grow on a chalcopyrite and a galena ore concentrate. When grown on the chalcopyrite concentrate, the bacteria exhibited a doubling time of 38.4 +/- 2.9 h, with a final cellular protein concentration of 185 mug/ml and solubilization of 10.3 g of copper per liter. When grown on the galena ore concentrate, the generation time was 39.6 +/- 2.7 h, with a final cellular protein concentration of 120 mug/ml. Galena was converted to lead salts soluble in 1 M ammonium acetate to a concentration of 20.2 g of lead per liter. X-ray diffraction and refractive-image analysis indicated that the smaller-sized particles were favored in this process. Galena was converted to anglesite, and soluble copper was liberated from chalcopyrite with the concurrent formation of jarosite. PMID- 16345531 TI - Biology of azospirillum-sugarcane association: enhancement of nitrogenase activity. AB - Azospirillum brasilense was reisolated from associations with callus tissue cultures of sugarcane and compared with stock cultures of the inoculated bacterium and related strains. Although the reisolate had a growth rate similar to stock cultures, it exhibited a severalfold increase in maximum specific activity of nitrogenase. The reisolate and the parent culture had similar ultrastructure. The general ultrastructure of Azospirillum is described. The bacterium was capsulated when grown on nitrogen-free nutrient agar plates and on callus, but was not capsulated when growing in a subsurface zone in N-free semisolid nutrient agar, except rarely in aging cultures. Capsulation may be a protective mechanism against unfavorable pO(2) under dinitrogen-fixing conditions. Pleomorphism occurred in capsulated forms, and the ultrastructure of these forms is described. PMID- 16345532 TI - Spore Load of Ascosphaera Species on Emerging Adults of the Alfalfa Leafcutting Bee, Megachile rotundata. AB - The spore load of Ascosphaera species spores on larval chalkbrood cadavers and newly emergent adults of the alfalfa leafcutting bee, Megachile rotundata, was determined. The spore content of chalkbrood cadavers ranged from 3 x 10 to 5 x 10. Adults emerging through zero to nine cadavers carried spores on all body parts examined by scanning electron microscopy. Estimates of the total number of spores obtained from a series of adult washes ranged from 9 x 10 to 8 x 10. Some adult males which emerged through no cadavers carried 10 to 10 spores, indicating that nesting materials might also have been contaminated. However, the control of chalkbrood in commercial bee populations may not be accomplished simply by providing clean nesting materials as adults may still emerge through diseased larvae. PMID- 16345533 TI - Influence of seasonal temperature on the temperature optima of bacteria in sediments of lake george, new york. AB - Temperature optima for the heterotrophic utilization of glucose and an amino acid mixture were determined throughout the year in sediments from Lake George, N.Y. The temperature optimum decreased with decreasing in situ temperature in the fall and winter, suggesting that selection for or adaptation by a psychrotrophic bacterial population occurred. Replicate plating of bacterial isolates from 3 and 20 degrees C indicated that a psychrotrophic bacterial population was present in the sediments throughout the year. These results indicate that decomposition and nutrient cycling processes in the sediments within the littoral zone of Lake George were probably not completely inhibited by winter temperatures, although process rates were decreased. PMID- 16345534 TI - Beta-Amylase Production by Bacillus polymyxa on a Corn Steep-Starch-Salts Medium. AB - Bacteria were found that are capable of producing good yields of beta-amylase in unrefined media. The culture filtrates are free of alpha-amylase and isoamylase. PMID- 16345535 TI - Carbon and electron flow in mud and sandflat intertidal sediments at delaware inlet, nelson, new zealand. AB - An investigation of carbon and electron flow in mud and sandflat intertidal sediments showed that the terminal electron acceptor was principally sulfate and that the carbon flow was mainly to CO(2). Studies with thin layers of sediment exposed to H(2) showed that methane production accounted for virtually none of the H(2) utilized, whereas sulfate reduction accounted for a major proportion of the gas uptake. At all sampling sites except one (site B7), rates of methanogenesis were low but sulfate concentrations in the interstitial water were high (>18 mM). At site B7, the sulfate concentrations declined with depth from 32 mM at 2 cm to <1 mM at 10 cm or below, and active methanogenesis occurred in the low-sulfate zone. Sulfate-reducing activity at this site initially decreased and then increased with depth so that elevated rates occurred in both the active and nonactive methanogenic zones. The respiratory index (RI) [RI = CO(2)/(CO(2) + CH(4))] for [2-C]acetate catabolism at site B7 ranged from 0.98 to 0.2 in the depth range of 2 to 14 cm. Addition of sulfate to sediment from the low-sulfate zone resulted in an increase in RI and a decrease in methanogenesis. At all other sites examined, RI ranged from 0.97 to 0.99 and was constant with depth. The results suggested that although methanogenesis was inhibited by sulfate (presumably through the activity of sulfate-reducing bacteria), it was not always limited by sulfate reduction. PMID- 16345536 TI - Microbial production of 4,4'-dihydroxybiphenyl: biphenyl hydroxylation by fungi. AB - Of 15 species of fungi examined for their ability to hydroxylate biphenyl, 10 produced 4-hydroxybiphenyl. Seven of the 10 also produced 4,4'-dihydroxybiphenyl. The most efficient strains, Absidia pseudocylindrospora NRRL 2770 and Absidia sp. NRRL 1341, were more closely examined to determine their growth characteristics and the kinetics of biphenyl hydroxylation in batch fermentation. In the absence of biphenyl, A. pseudocylindrospora 2770 and Absidia sp. 1341 grew about as rapidly and efficiently in a defined glucose minimal medium as in a complex medium. Substrate yield coefficients for glucose in both media were 0.4 to 0.5 g of biomass per g of glucose, and the specific growth rate was about 0.17 h (doubling time, about 4 h). In this unoptimized system, 10 to 15 g of biomass per liter (dry weight) could be produced, using a defined salt solution and glucose as sole carbon and energy source. In the presence of biphenyl, growth was inhibited, more so for strain 1341 than for strain 2770. However, the specific activity for biphenyl hydroxylation (milligrams of biphenol per gram of biomass) was about 3.5 times greater for strain 1341. Furthermore, biphenyl hydroxylation appeared to require the presence of an oxidizable carbon and energy source (and perhaps growth) to proceed and, at least for strain 1341, hydroxylation seemed to be more efficient in the complex medium. PMID- 16345537 TI - Microbial Oxidation of Gaseous Hydrocarbons: Production of Secondary Alcohols from Corresponding n-Alkanes by Methane-Utilizing Bacteria. AB - Over 20 new strains of methane-utilizing bacteria were isolated from lake water and soil samples. Cell suspensions of these and of other known strains of methane utilizing bacteria oxidized n-alkanes (propane, butane, pentane, hexane) to their corresponding secondary alcohols (2-propanol, 2-butanol, 2-pentanol, 2-hexanol). The product secondary alcohols accumulated extracellularly. The rate of production of secondary alcohols varied with the organism used for oxidation. The average rate of 2-propanol, 2-butanol, 2-pentanol, and 2-hexanol production was 1.5, 1.0, 0.15, and 0.08 mumol/h per 5.0 mg of protein in cell suspensions, respectively. Secondary alcohols were slowly oxidized further to the corresponding methylketones. Primary alcohols and aldehydes were also detected in low amounts (rate of production were 0.05 to 0.08 mumol/h per 5.0 mg of protein in cell suspensions) as products of n-alkane (propane and butane) oxidation. However, primary alcohols and aldehydes were rapidly metabolized further by cell suspensions. Methanol-grown cells of methane-utilizing bacteria did not oxidize n alkanes to their corresponding secondary alcohols, indicating that the enzymatic system required for oxidation of n-alkanes was induced only during growth on methane. The optimal conditions for in vivo secondary alcohol formation from n alkanes were investigated in Methylosinus sp. (CRL-15). The rate of 2-propanol and 2-butanol production was linear for the 40-min incubation period and increased directly with cell protein concentration up to 12 mg/ml. The optimal temperature and pH for the production of 2-propanol and 2-butanol were 40 degrees C and pH 7.0. Metalchelating agents inhibited the production of secondary alcohols. The activities for the hydroxylation of n-alkanes in various methylotrophic bacteria were localized in the cell-free particulate fractions precipitated by centrifugation between 10,000 and 40,000 x g. Both oxygen and reduced nicotinamide adenine dinucleotide were required for hydroxylation activity. The metal-chelating agents inhibited hydroxylation of n-alkanes by the particulate fraction, indicating the involvement of a metal-containing enzyme system in the oxidation of n-alkanes. The production of 2-propanol from the corresponding n-alkane by the particulate fraction was inhibited in the presence of methane, suggesting that the subterminal hydroxylation of n-alkanes may be catalyzed by methane monooxygenase. PMID- 16345538 TI - Microbial Oxidation of Gaseous Hydrocarbons: Production of Methylketones from Corresponding n-Alkanes by Methane-Utilizing Bacteria. AB - Cell suspensions of methane-utilizing bacteria grown on methane oxidized n alkanes (propane, butane, pentane, hexane) to their corresponding methylketones (acetone, 2-butanone, 2-pentanone, 2-hexanone). The product methylketones accumulated extracellularly. The rate of production of methylketones varied with the organism used for oxidation; however, the average rate of acetone, 2 butanone, 2-pentanone, and 2-hexanone production was 1.2, 1.0, 0.15, and 0.025 mumol/h per 5.0 mg of protein in cell suspensions. Primary alcohols and aldehydes were also detected in low amounts as products of n-alkane (propane and butane) oxidation, but were rapidly metabolized further by cell suspensions. The optimal conditions for in vivo methylketone formation from n-alkanes were compared in Methylococcus capsulatus (Texas strain), Methylosinus sp. (CRL-15), and Methylobacterium sp. (CRL-26). The rate of acetone and 2-butanone production was linear for the first 60 min of incubation and directly increased with cell concentration up to 10 mg of protein per ml for all three cultures tested. The optimal temperatures for the production of acetone and 2-butanone were 35 degrees C for Methylosinus trichosporium sp. (CRL-15) and Methylobacterium sp. (CRL-26) and 40 degrees C for Methylcoccus capsulatus (Texas). Metal-chelating agents inhibited the production of methylketones, suggesting the involvement of a metal containing enzymatic system in the oxidation of n-alkanes to the corresponding methylketones. The soluble crude extracts derived from methane-utilizing bacteria contained an oxidized nicotinamide adenine dinucleotide-dependent dehydrogenase which catalyzed the oxidation of secondary alcohols. PMID- 16345539 TI - Occurrence of cytophagas in sewage plants. AB - With the application of plate count methods and of the KOH-flexirubin test, bacteria belonging to the Cytophaga group were proved to occur regularly in samples from biological sewage treatment facilities. Generally, the percentage of Cytophaga colonies of the total heterotrophic colonies was lowest in the inflow sewage water as compared with the values found in activated sludge, trickling filter, and effluent samples. During an observation period of 16 months, the highest percentages of cytophagas were found in winter samples from activated sludge and trickling filters. Furthermore, cytophagas were shown to have high percentages of the bacteria lytic to polymeric substrates such as cellulose, chitin, dextran, pectin, xylan, and gelatin. Thus, it is suggested that cytophagas may contribute to sewage purification, especially at cold temperatures and by polymer breakdown. Cytophaga strains isolated were shown to have gliding motility, flexirubin pigmentation, and a low guanine plus cytosine base ratio in common. The strains were roughly subdivided into a spreading, a nonspreading, and a cellulolytic group. PMID- 16345540 TI - Production of Eremofortins A, B, and C Relative to Formation of PR Toxin by Penicillium roqueforti. AB - The production of various eremophilane-type sesquiterpenes by Penicillium roqueforti strains has allowed us to propose a biochemical pathway for PR toxin synthesis. A time-course study of P. roqueforti metabolite production by high performance liquid chromatography was performed to check this hypothetical pathway. The results obtained suggested that eremofortin C was the direct precursor of PR toxin in the P. roqueforti cell. Attempts to determine the amount of PR toxin in the mycelium failed. It was shown that the absence of PR toxin in mycelium was due to its instability during the extraction procedure. PMID- 16345541 TI - Alteration of Acrylonitrile-Methylacrylate-Butadiene Terpolymer by Nocardia rhodochrous and Penicillium notatum. AB - [C]Barex-210, a terpolymer of acrylonitrile, methylacrylate, and butadiene, was tested for bioconversion. Powdered samples of polymer, each specifically C labeled at different carbon atoms of the polymer, were incubated with either Nocardia rhodochrous or Penicillium notatum in an enriched growth medium for various periods of time. After 6 months of incubation, the C-labeled polymer was transformed from a high-molecular-weight material completely soluble in dimethyl formamide (DMF) into both a lower-molecular-weight form still soluble in DMF and a second form that was no longer soluble in DMF. The amount of C-labeled carbon atoms converted into DMF-insoluble material was 8% of the backbone carbon-carbon atoms and 12% of the side-chain nitrile and acrylate atoms from the acrylonitrile methylacrylate copolymer and 60% of the elastomer (acrylonitrile-butadiene copolymer) atoms. Metabolism of the polymer was not established from measurements of metabolic CO(2). Evolution of CO(2) amounted to only 0.3, 0.6, 1.8, and 3.3% of these four fractions, respectively. Although the transformation of high molecular-weight polymer into DMF-insoluble material was rapid in the early stages of microbial growth, the accompanying CO(2) evolution was much slower. Further evidence of polymer alteration was indicated by the infrared spectrum of the insoluble material, which showed a disappearance of the nitrile and methylacrylate peaks. PMID- 16345542 TI - Temperature compensation of [u-C]glucose incorporation by microbial communities in a river with a fluctuating thermal regime. AB - In summer, the river Saar in the southwest of Germany exhibits distinct temperature fluctuations from 8 degrees C at the source to nearly 30 degrees C in the middle region. Temperature optima for bacterial plate counts and the uptake velocity of [U-C]glucose by the natural microbial communities of different regions ranged from 20 to 30 degrees C, which is significantly above the mean annual water temperature. A correlation between temperature optima and different seasons or habitats was not observed. Despite the relatively high temperature optima, the turnover time for glucose was shortest at temperatures around the mean annual water temperature, due to changes in the substrate affinity. At limiting substrate concentrations, the higher substrate affinity at lower temperatures may lead to a higher real activity at in situ temperatures, and a compensatory stabilization of uptake rates at fluctuating temperatures is possible. PMID- 16345543 TI - Interrelationships between microbiological and chemical parameters of sandy beach sediments, a summer aspect. AB - At 12 stations located in sandy beach sediments of the brackish water Kiel Fjord and Kiel Bight (Baltic Sea, FRG), variations and interrelationships of microbiological, chemical, and physicochemical parameters were monitored. Depending upon location, wide variations of a number of parameters reflecting dissolved organic and inorganic nutrients, chlorophyll a, microbial number, and uptake activity of glucose were measured. Whereas most of the parameters generally showed the tendency to decrease from the inner to the outer Kiel Fjord, individual parameters (oxygen, particulate nitrogen, ribose, chlorophyll a, glucose/fructose ratio) increased with increasing distance from the inner Kiel Fjord. Similarities in the local variation pattern demonstrated various relationships between individual parameters. Among those, dissolved organic nutrients on the one hand and inorganic nutrients on the other hand were closely linked together. Variations of organic and inorganic nutrients corresponded to variations of microbial activity and physicochemical parameters. By comparing standing stock carbon with carbon production, a microbial biomass turnover time of about 100 h was calculated. Approximately 50% of the microphytobenthos primary production was fixed by microorganisms. Daily microbial carbon production (43 mg of C per m) was in the range of meiofauna carbon (35 mg of C per m). PMID- 16345544 TI - Hot Acidified Cupric Acetate Soaks for Eradication of Xanthomonas campestris from Crucifer Seeds. AB - Acidified cupric acetate soaks were tested for eradication of Xanthomonas campestris from naturally infected crucifer seeds. The pathogen was eradicated from seeds by soaking in 0.5% cupric acetate dissolved in 0.005 N acetic acid for 20 min at 35, 40, 45, and 50 degrees C but not 25 degrees C. Moreover, normal bacterial flora of crucifer seeds and the seed-borne Phoma lingam and Alternaria spp. were reduced by 95, 92, and 81%, respectively, after the cupric acetate treatment at 40 degrees C. The seed germination percentage was generally reduced, but the amount of reduction depended upon the treatment temperature and plant cultivar. At 50 degrees C, less than 50% of the seed of all 12 cultivars tested germinated, whereas at 40 degrees C more than 50% of the seeds of most cultivars germinated. Treating seeds in cupric acetate at 40 degrees C should prove useful for eradicating X. campestris from seeds of breeding lines and stock seed used for hybrid seed production. Furthermore, a significant reduction in total bacterial flora and seed-borne fungi suggests the usefulness of the treatment for other microorganisms associated with other seeds or foodstuffs. PMID- 16345545 TI - Relationship between the sterol content of yeast cells and their fermentation activity in grape must. AB - In grape must of high sugar concentration, yeast growth, the viability rate of "resting" yeast cells, and fermentation activity were stimulated under certain conditions of aeration and temperature. This stimulation might be interpreted as being a result of the yeast cell sterol content. The addition of certain sterols to the fermenting medium was able to increase this sterol content. According to aeration conditions of the medium, which determined the sterol content of yeasts, the sterols added in the medium acted as (i) growth factors, (ii) fermentation inhibitors, and (iii) survival factors for the yeast. PMID- 16345546 TI - Formation and Purification of Serratia marcescens Arylsulfatase. AB - The effects of culture conditions on arylsulfatase production by six strains of the genus Serratia were studied. Synthesis of arylsulfatases in all six strains was repressed in media with inorganic sulfate or methionine as the sole source of sulfur and derepressed by the addition of tyramine. Serratia marcescens IFO 3046 grew most rapidly and produced a high level of arylsulfatase when cultured on mannitol with inorganic sulfate and tyramine. The derepressed synthesis of arylsulfatase in S. marcescens was not subject to strong catabolite repression. The molecular weight of purified arylsulfatase was determined to be between 46,000 and 49,000. Arylsulfatase from S. marcescens differed in K(m) and V(max) values, substrate specificities, fluoride inhibition, and electrophoretic mobility from the enzyme from K. aerogenes, but had the same molecular weight as the latter. PMID- 16345547 TI - Nitrite and nitrous oxide accumulation during denitrification in the presence of pesticide derivatives. AB - Temporary accumulation of nitrite and nitrous oxide was observed in soil incubated under anaerobic conditions when derivatives of the insecticide chlordimeform [(N-4-chloro-o-tolyl)-N',N' -dimethylformamidine] were added. Chlordimeform did not affect the denitrification process, but N-formyl-4-chloro-o toluidine and 4-chloro-o-toluidine caused an inhibition as determined by the accumulation of nitrite and nitrous oxide. A simultaneous application of the insecticide and its derivatives resulted in a stronger inhibitory effect than the application of each compound separately. Aniline intermediates of other pesticides also inhibited denitrification in soil, and they proved to be more effective than their parent compound. PMID- 16345548 TI - Deoxyribonucleic Acid Repair in Bacillus subtilis: Development of Competent Cells into a Tester for Carcinogens. AB - The development of competent transformed Bacillus subtilis into a tester system for carcinogens is described. Precocious or noninduced activation of SOS functions occur in competent cells. Thus, lower doses or concentrations of SOS inducing agents are needed to cause cell death due to indigenous prophage activation and lysis of bacteria. The two known defective prophages in B. subtilis enhance the sensitivity of competent cells to the carcinogens ultraviolet light, mitomycin C, and methyl methanesulfonate. However, these same cells have no enhanced sensitivity for the non-carcinogenic ethyl methanesulfonate or for nalidixic acid. Therefore, competent B. subtilis appear to be a sensitive tester for carcinogens. PMID- 16345549 TI - Cellulose Metabolism by the Termite Flagellate Trichomitopsis termopsidis. AB - The end products of cellulose metabolism by the trichomonad flagellate Trichomitopsis termopsidis from the termite Zootermopsis sp. were investigated by growing axenic flagellates on [C]cellulose. The growth of T. termopsidis resulted in the release of label into the supernatant fraction of the culture fluid, and > 75% was volatile under acid conditions. The label was analyzed for CO(2) and for [C]volatile compounds by vacuum distillation under acid and alkaline conditions in disposable micro-distillation vessels. The distillate and undistilled culture supernatant fluid were chromatographed on cellulose thin layers to identify the labeled end product. T. termopsidis produced CO(2) and [C]acetate which accounted for 25 to 30% and 55 to 60% of the labeled end products, respectively. The ratio of label in CO(2) to acetate suggests that they are produced in equimolar amounts. No neutral volatile compounds were produced. The remaining unidentified end product (10 to 20%) was not volatile nor extractable into ether. Hydrogen was produced by T. termopsidis, and the cells were killed by the drug metronidazole. Enzymatic activities were found which account for these end products: pyruvate:ferredoxin oxidoreductase and hydrogenase. The results indicate that acetate is the end product of T. termopsidis cellulose metabolism and is available to the termite for energy metabolism and biosynthesis. PMID- 16345550 TI - Populations of methane-producing bacteria and in vitro methanogenesis in salt marsh and estuarine sediments. AB - Most probable numbers (MPNs) of methanogens in various salt marsh and estuarine sediments were determined with an anaerobic, habitat-simulating culture medium with 80% H(2) plus 20% CO(2) as substrate. Average MPNs for the short Spartina (SS) marsh sediments of Sapelo Island, Ga., were maximal at the 5- to 7-cm depth (1.2 x 10/g of dry sediment). Populations decreased to approximately 880/g of dry sediment at the 34- to 36-cm depth. There was no significant difference between summer and winter populations. In tall Spartina (TS) marsh sediments, average populations were maximal (1.2 x 10/g of dry sediment) in the upper 0- to 2-cm zone; populations from the 5- to 36-cm zones were similar (average of 9 x 10/g of dry sediment). Methanogenic populations for TS sediments of James Island Creek marsh, Charleston, S.C., were similar (average of 3 x 10/g of dry sediment) for all depths tested (0 to 22 cm), which was comparable to the trend observed for TS sediments at Sapelo Island, Ga. Sediment grab samples collected along a transect of James Island Creek and its adjacent Spartina marsh had MPNs that were approximately 20 times greater for the region of Spartina growth (average of 10/g of dry sediment) compared with the channel (approximately 5 x 10 methanogens per g of dry sediment). A similar trend was found at Pawley's Island marsh, S.C., but populations were approximately one order of magnitude lower. In vitro rates of methanogenesis with SS sediments incubated under 80% H(2)-20% CO(2) showed that the 5- to 7-cm region exhibited maximal activity (51 nmol of CH(4) g h), which was greater than rates for sediments above and below this depth. SS sediment samples (5 to 7 cm) incubated under 100% N(2) and supplemented with formate exhibited rates of methanogenesis similar to those generated by samples under 80% H(2)-20% CO(2). Replacing the N(2) atmosphere with H(2) resulted in an eightfold decrease in the rate of methanogenesis. In vitro methanogenic activity by TS salt marsh sediments, incubated under 80% H(2)-20% CO(2), was similar for all depths tested (0 to 22 cm). TS sediment samples (0 to 7 cm) supplemented with formate and incubated under 100% N(2) had greater rates of methanogenesis compared with unsupplemented samples. PMID- 16345551 TI - Tracer analysis of methanogenesis in salt marsh soils. AB - Differences in paths of carbon flow have been found in soils of the tall (TS) and short (SS) Spartina alterniflora marshes of Sapelo Island, Ga. Gaseous end products of [U-C]glucose metabolism were CO(2) and CH(4) in the SS region and primarily CO(2) in the TS region. Sulfate concentration did not demonstrably affect glucose catabolism or the distribution of end products in either zone. [U C]acetate was converted to CO(2) and CH(4) in the SS soils and almost exclusively to CO(2) in the TS soils. Sulfate concentration did not affect acetate metabolism in the SS soils; however, a noticeable effect of sulfate dilution was seen in TS soils. Sulfate dilution in TS samples resulted in increased methane formation. Total glucose and acetate metabolism were similar in TS and SS soils despite differences in end products. A microbial community characterized by fermentative/sulfate-reducing processes has developed in TS soils as opposed to the fermentative/methanogenic/sulfate-reducing community found in SS soils. PMID- 16345553 TI - Microbial colonization and decomposition of carex litter in an arctic lake. AB - The decomposition and microbial colonization of Carex leaf litter were examined in an arctic lake in Alaska during the summer of 1978. Dried leaf segments in screen bags were placed at various locations and depths for 13 and 26 days. Weight loss varied from 24.15 to 33.56% and from 27.69 to 65.01% after 13 and 26 days, respectively. Abiotic controls lost approximately 19.5% with no subsequent change. Weight loss significantly correlated with microbial colonization as measured by alkaline phosphatase activity (r = 0.780), cellulase activity (r = 0.613), heterotrophic CO(2) fixation (r = 0.835), and acetate incorporation into microbial lipids (r = 0.618). Alkaline phosphatase activity correlated with cellulase activity (r = 0.889), and heterotrophic CO(2) fixation correlated with acetate incorporation into lipids (r = 0.712). Weight loss after 26 days inversely correlated with the logarithm of the depth of incubation regardless of whether incubation occurred on the sediment surface or in the water column. These findings suggest that a rapid initial period of microbial colonization is driven by nutrients derived from the litter and that the rate of these processes is controlled by a factor(s) inversely related to the logarithm of depth, such as light intensity, primary production, or turbulence. PMID- 16345552 TI - Penitrem A and Roquefortine Production by Penicillium commune. AB - Extracts of Penicillium commune, a fungus isolated from cottonseed, showed biological activity in day-old cockerels. Two neurotoxic metabolites were isolated and identified as penitrem A and roquefortine. This is the first report of roquefortine being produced by a fungus other than Penicillium roqueforti as well as the first report of penitrem A and roquefortine being produced in the same culture. Production of these toxins on liquid media and cottonseed was determined. PMID- 16345554 TI - Enrichment and association of bacteria and particulates in salt marsh surface water. AB - Elevated counts of bacteria were found during outgoing tides in surface microlayers ( approximately 300 mum) of Sippewissett salt marsh, Falmouth, Massachusetts, and Palo Alto salt marsh, Palo Alto, California. At both sampling sites, the degrees by which bacteria were concentrated into the surface microlayer were linearly dependent upon surface concentration of particulate material. A significant percentage of bacteria in the microlayer were found to be attached to particulate material, while bacterial populations in the subsurface water were largely planktonic. Proportions of the bacterial populations which could be grown on seawater nutrient agar were also greater in the microlayer than in the subsurface waters and were positively correlated with the fraction of bacteria attached to particulate matter. Data from these studies suggest that particulates in the microlayer waters of the salt marsh influenced the observed increase in both the readily grown and the total numbers of bacteria. PMID- 16345555 TI - Polyacrylamide slab gel electrophoresis of soluble proteins for studies of bacterial floras. AB - A polyacrylamide slab gel electrophoresis procedure was used to compare cellular proteins from bacterial isolates of gingival crevice floras. Isolates with identical protein patterns consistently were shown to be members of the same species. When used to screen isolates, the procedure reduced total analytical time and expense without sacrificing accuracy, and it provided additional verification of the identity of strains characterized by conventional phenotypic tests. PMID- 16345556 TI - Microbial production of pectin from citrus peel. AB - A new method for the production of pectin from citrus peel was developed. For this purpose, a microorganism which produces a protopectin-solubilizing enzyme was isolated and identified as a variety of Trichosporon penicillatum. The most suitable conditions for the pectin production were determined as follows. Citrus (Citrus unshiu) peel was suspended in water (1:2, wt/vol), the organism was added, and fermentation proceeded over 15 to 20 h at 30 degrees C. During the fermentation, the pectin in the peel was extracted almost completely without macerating the peel. By this method, 20 to 25 g of pectin was obtained per kg of peel. The pectin obtained was special in that it contained neutral sugar at high levels, which was determined to have a molecular weight suitable for practical applications. PMID- 16345557 TI - Immunofluorescent Assay for the Marine Ammonium-Oxidizing Bacterium Nitrosococcus oceanus. AB - Nitrification is one of the important microbiological transformations of nitrogen in the ocean. Traditional enrichment-culture methods for enumerating the autotrophic bacteria which oxidize ammonium to nitrite are very time consuming (months) and are believed to seriously underestimate natural abundances. A fluorescent-antibody assay for a marine ammonium-oxidizing bacterium was developed to provide a rapid and direct means of identifying these microorganisms. Antibodies to Nitrosococcus oceanus were prepared and tested against pure cultures of marine, freshwater, and soil ammonium oxidizers and against bacteria from natural seawater samples. Cell counts of culture samples determined by the fluorescent-antibody assay agreed with hemacytometer and acridine orange counts. Our results demonstrated that the immunofluorescent assay is a powerful tool for the detection of Nitrosococcus in the marine environment. PMID- 16345558 TI - New Method of Producing Protoplasts of Aureobasidium pullulans. AB - A rapid and relatively inexpensive method for producing protoplasts of the black yeast Aureobasidium pullulans is described. The procedure involves anaerobic incubation with the lytic preparation Driselase. PMID- 16345559 TI - Improved Medium for Detection of Citrate-Fermenting Streptococcus lactis subsp. diacetylactis. AB - A modified medium which distinguished between citrate-fermenting and non-citrate fermenting species of lactic streptococci within 48 h was developed. In addition, the occurrence of citrate-negative variants in citrate-positive populations of Streptococcus lactis subsp. diacetylactis could be detected. PMID- 16345560 TI - Stability of the Plasma Membrane in Saccharomyces rouxii and Its Relationship to Glucose Tolerance. AB - The stability of spheroplasts from the osmotrophic yeast Saccharomyces rouxii was studied in buffered solutions of mannitol and glucose. The plasma membranes from cells grown in high glucose concentrations were more stable to osmotic lysis than were membranes from cells grown in lower glucose concentrations. Mannitol was a better osmotic stabilizer than glucose, except when the cells were grown in a high glucose concentration. Spheroplasts from a glucose tolerant-deficient mutant were much less stable than the corresponding spheroplasts from the parent strain, especially when suspended in glucose solutions. These results suggest an involvement of the plasma membrane in the glucose-tolerant mechanism of S. rouxii. PMID- 16345561 TI - Silica gel plates for culture of marine and nonmarine organisms. AB - Marine and nonmarine species of bacteria grew well when a simplified silica gel plate preparation was used. PMID- 16345563 TI - Dispersal of Aspergillus fumigatus from Sewage Sludge Compost Piles Subjected to Mechanical Agitation in Open Air. AB - Aerosolization of the thermophilous fungal opportunist Aspergillus fumigatus from mechanically agitated compost piles was examined at a pilot-scale sewage sludge composting facility and two other selected test sites. Aerosols of A. fumigatus downwind from stationary compost piles were insignificant in comparison with those downwind from agitated piles. These aerosols were generated by a front-end loader moving and dropping compost. Aerial concentrations of the fungus at distances downwind from the point of emission were used to determine an emission rate for A. fumigatus associated with the moving operations. The maximum emission rate, 4.6 x 10A. fumigatus particles per s, was used to calculate predicted concentrations in an unobstructed plume with restrictive, neutral, and dispersive atmospheric mixing conditions up to 1 km downwind from the emission source. PMID- 16345562 TI - High Cellobiase and Xylanase Production by Sclerotium rolfsii UV-8 Mutant in Submerged Culture. AB - Sclerotium rolfsii UV-8 mutant secretes high levels of cellobiase and xylanase in addition to having high cellulase production. The apparent K(m) and V(max) of cellobiase (grown in NM-2 + 2% corn steep liquor medium) with cellobiose as a substrate were 5.6 mM and 22.2 mumol of glucose liberated per min per ml of culture filtrate, respectively. The addition of 2% corn steep liquor to NM-2 medium increased endo-beta-glucanase, cellobiase, and xylanase yields by approximately 1.5-fold. PMID- 16345564 TI - Enumeration and relative importance of acetylene-reducing (nitrogen-fixing) bacteria in a delaware salt marsh. AB - Three groups of N(2)-fixing bacteria were enumerated from the top 1 cm of the surface in four vegetational areas in a Delaware salt marsh. The results over the 9-month sampling period showed that there were no discernible seasonal patterns for any of the groups enumerated (Azotobacter sp., Clostridium sp., and Desulfovibrio sp.). Azotobacter sp. was present in numbers of 10 per g of dry mud, whereas the two anaerobic fixers were present in much lower numbers (10 to 10 per g of dry mud). There were no differences in the numbers of each group among the different vegetational areas, indicating that there was a heterogeneous population of N(2) fixers present. Additional studies indicate that the activity of sulfate reducers (Desulfovibrio sp.) may account for as much as 50% of the total observed acetylene reduction activity. Oxygen was found to exert little effect on the observed acetylene reduction activity, indicating that stable aerobic and anaerobic microenvironments exist in the surface layer of marsh sediments. PMID- 16345565 TI - Death of Micrococcus luteus in Soil. AB - Micrococcus luteus cells died relatively quickly when they were added to natural soil. The results were similar for soil in nature and as soil samples in the laboratory. The cells died more quickly when nutrients were added to the soil. Those cells that survived soil residence exhibited a temporary lengthening of the time required for colonial growth and pigment formation on laboratory media. They had not gained increased survival capability, however. This was evident when they were retested in soil. Good survival of the M. luteus cells was noted when the soil was incubated at lowered temperatures. Some protection to the cells was provided by slow drying of the soil during incubation or by addition of NaCl. Microscopic examination of the soil revealed that the M. luteus cells were being physically destroyed and that two different bacteria were growing in the areas where the cells had lysed. It was suggested that bacterial predators in the soil might be associated with the death of the M. luteus cells. PMID- 16345566 TI - Bacterial Predators of Micrococcus luteus in Soil. AB - Micrococcus luteus cells died relatively rapidly when they were added to natural soil. Microscopic observation showed that the cells were being physically destroyed by bacterial predators in the soil. Two of these predators were responsible for the initial, main attack, and they were isolated. The isolates on laboratory media lysed M. luteus cells in a manner similar to the attacks that occurred in soil. Neither predator was obligate, however, nor were they nutritionally fastidious. One of these bacteria produced mycelium and conidia. Under nutritionally poor conditions it used slender filaments of mycelium to seek out host cells. It had at least some of the characteristics of a Streptoverticillium species. The other bacterium was a short, gram-negative rod that did not easily fit into any of the known groups of gram-negative bacteria. It attached to host cells, but its mechanism of lysing these cells is not known. PMID- 16345567 TI - Fluorescent Staining Technique for Nucleoid Regions of Streptosporangium albidum and Streptosporangium brasiliense. AB - Fluorescent staining procedures were developed for elucidating the nucleoid region in Streptosporangium albidum and Streptosporangium brasiliense. In these procedures, plugs of nutrient agar were inoculated with the microorganims and then covered with a sterile glass slide. The growing cells adhered to the surface of the slide and remained attached throughout the staining procedures. Two separate staining methods were utilized, one with bisbenzimid H33258 and the other with auramine O. Fluorescent microscopy revealed intensely stained nucleoid regions within mycelia, spores, and sporangia. PMID- 16345568 TI - Separation of d-(+)-Nicotine from a Racemic Mixture by Stereospecific Degradation of the l-(-) Isomer with Pseudomonas putida. AB - Incubation of racemic mixtures of dl-(+/-)-nicotine with Pseudomonas putida resulted in a complete stereoselective degradation of the l-(-) isomer. Unnatural d-(+)-nicotine, which is of pharmacological interest for stereochemical studies of various nicotine-responsive systems, was not affected by the bacterium and was recovered by extraction. PMID- 16345569 TI - Isolation of Methanobacterium bryantii from a Deep Aquifer by Using a Novel Broth Antibiotic Disk Method. AB - Methanobacterium bryantii was isolated from a mixed-culture enrichment of a water sample from a deep aquifer by using a complex growth medium supplemented with antibiotic susceptibility disks to inhibit the growth of non-methanogenic bacteria. PMID- 16345570 TI - Acetylene reduction assay for nitrogenase activity: gas chromatographic determination of ethylene per sample in less than one minute. AB - Ammoniacal silver nitrate (10 mg/ml) was added to terminate acetylene reduction assays used to measure nitrogenase activity. Silver nitrate quantitatively precipitated acetylene as the carbide salt, but did not affect the ethylene formed. The vials containing ethylene can be analyzed gas chromatographically at the rate of about 13 samples in 10 min. PMID- 16345571 TI - Blue Light Inhibits Mycotoxin Production and Increases Total Lipids and Pigmentation in Alternaria alternata. AB - [This corrects the article on p. 1075 in vol. 38.]. PMID- 16345572 TI - Culture of five commonly used Acid-producing bacteria on banana pulp. AB - A significant fraction of the bananas and plantains produced in the humid tropics are wasted due to rapid spoilage. The use of acid-producing, anaerobic fermentation may be a safe and economical way to preserve the fruit in both large and small-scale applications. The abilities of five commonly used, acid producing bacteria to ferment media containing ripe or green bananas were examined. Three homofermentative bacteria, Lactobacillus bulgaricus, Streptococcus thermophilus, and Streptococcus faecalis, typically reduced the pH of a puree of ripe bananas from 4.8 to about 3.5 to 3.6 in 7 days at 37 degrees C. Action on green bananas was similar, but the drop in pH was less pronounced. Very little gas evolved. Lactobacillus fermentum (formerly Lactobacillus fermenti) and Leuconostoc mesenteroides showed active gas evolution and a significant drop in pH. About twice as much gas evolved from ripe banana medium as from green bananas. The fermented purees were spongy, soft solids floating in a clear liquor. The color was medium to light creamy brown, and the odor was pleasant. Fermented products stored at room temperature retained good appearance and aroma throughout a 1-month storage test. PMID- 16345573 TI - Sulfhydrolase activity in sediments of wintergreen lake, kalamazoo county, michigan. AB - The hydrolysis of p-nitrophenyl sulfate, p-nitrocatechol sulfate, and [S]sodium dodecyl sulfate was examined in anoxic sediments of Wintergreen Lake, Michigan. Significant levels of sulfhydrolase activity were observed in littoral, transition, and profundal sediment samples. Rates of sulfate formation suggest that the sulfhydrolase system would represent a major source of sulfate within these sediments. Sulfate formed by ester sulfate hydrolysis can support dissimilatory sulfate reduction as shown by the incorporation of S from labeled sodium dodecyl sulfate into H(2)S. Sulfhydrolase activity varied with sediment depth, was greatest in the littoral zone, and was sensitive to the presence of oxygen. Estimations of ester sulfate concentrations in sediments revealed large quantities of ester sulfate ( approximately 30% of total sulfur). Both total sulfur and ester sulfate concentrations varied with the sediment type and were two to three orders of magnitude greater than the inorganic sulfur concentration. PMID- 16345574 TI - Method for Testing Degree of Infectivity of Rhizobium meliloti Strains. AB - The infectiveness of different strains of Rhizobium meliloti was tested with a technique that uses the addition of tetracycline to the root medium. To stop the infection, the antibiotic was added some time after the inoculation of Medicago sativa plants. A coefficient of infectivity for each strain was calculated according to the number of nodules that appeared with and without the addition of the antibiotic. This method seems useful in infectivity studies and is simpler and easier to perform than the test of competence between strains. PMID- 16345575 TI - Distribution and identification of luminous bacteria from the sargasso sea. AB - Vibrio fischeri and Lucibacterium harveyi constituted 75 of the 83 luminous bacteria isolated from Sargasso Sea surface waters. Photobacterium leiognathi and Photobacterium phosphoreum constituted the remainder of the isolates. Luminescent bacteria were recovered at concentrations of 1 to 63 cells per 100 ml from water samples collected at depths of 160 to 320 m. Two water samples collected at the thermocline yielded larger numbers of viable, aerobic heterotrophic and luminous bacteria. Luminescent bacteria were not recovered from surface microlayer samples. The species distribution of the luminous bacteria reflected previously recognized growth patterns; i.e., L. harveyi and V. fischeri were predominant in the upper, warm waters (only one isolate of P. phosphoreum was obtained from surface tropical waters). PMID- 16345576 TI - Acetate as sole carbon and energy source for growth of methanosarcina strain 227. AB - Methanosarcina strain 227 grew rapidly and produced methane on a mineral medium containing acetate as the sole added organic substrate. Cell yields but not doubling times were affected by the presence or absence of yeast extract. Greater cell yields occurred in yeast extract medium than in mineral medium. Radioactive labeling studies showed that acetate was decarboxylated in mineral medium, as was shown previously in complex medium. The specific radioactivity of methane produced per specific acitvity of acetate added was not significantly different in yeast extract medium compared with mineral medium. Unequivocal evidence indicates that the cleavage of acetate to methane and carbon dioxide provided the energy for growth in the presence or absence of other organic compounds; these latter compounds do not serve as energy sources, electron donors, or significant sources of methane during this aceticlastic reaction. PMID- 16345577 TI - Bacterioplankton secondary production estimates for coastal waters of british columbia, antarctica, and california. AB - The principal objective of this study was to quantify the rate of heterotrophic bacterioplankton production. Production was estimated by two approaches: (i) measurement of increasing bacterial abundance with time in filtered (3-mum pore size) seawater and (ii) estimation of bacterial deoxyribonucleic acid synthesis by tritiated thymidine incorporation in unfractionated seawater. The two approaches yielded comparable results when used at the Controlled Ecosystem Population Experiment (Saanich Inlet, British Columbia, Canada), at McMurdo Sound (Antarctica), and off Scripps Pier (La Jolla, Calif.). Estimated bacterioplankton production was lower in Antarctic samples (ranging from approximately 0 to 2.9 mug of C liter day) than in those from the other two sites (ranging from 0.7 to 71 mug of C liter day). In all three regions studied, it appeared that a significant fraction of the total primary production was utilized by the bacterioplankton and that substantial growth could occur in the absence of large particles. These results support the conclusion that bacterioplankton are a quantitatively important component of coastal marine food webs. PMID- 16345578 TI - Accumulation of Soybean Lectin-Binding Polysaccharide During Growth of Rhizobium japonicum as Determined by Hemagglutination Inhibition Assay. AB - A hemagglutination inhibition assay was used to estimate the presence of soybean lectin-binding polysaccharide in whole culture, culture supernatant, and isolated exopolysaccharide of Rhizobium japonicum USDA 138. The occurrence of 0.1 to 0.2 mug of lectin-binding polysaccharide could be detected within 2 h with a 0.5-ml total sample. Lectin-binding polysaccharide was detected in all preparations during both exponential and stationary growth phases. The formation of lectin binding polysaccharide was not, whereas that of total exopolysaccharide was, markedly affected by culture conditions. PMID- 16345579 TI - Production of aseptic spores of vesicular-arbuscular endophytes and their viability after chemical and physical stress. AB - The survival of germinating spores of vesicular-arbuscular endophytes after treatments with oxidizing agents, antibiotics, moist heat, ultrasonic radiation, and ultraviolet radiation was compared with that of their contaminating microbes. Spores of three species were rapidly decontaminated by treatment with 0.42% (wt/vol) chlorine available from 5.0% (wt/vol) chloramine-T at 30 degrees C for 20 to 40 min depending on the species and the soil from which they were extracted. This treatment did not change spore viability. The survival of spores was reduced by exposure for 20 min to 1.11% chlorine at 30 degrees C for Glomus caledonius or at 35 degrees C for Acaulospora laevis. Growth of any bacteria surviving treatment with oxidizing agents was inhibited by 100 mug of chloramphenicol per ml in agar; however, spore germination and germ tube growth were reduced only by concentrations greater than 200 mug/ml in agar. Spore germination was decreased by concentration of pimaracin, which controlled fungal growth. The spores survived moist heat at 40 degrees C for 80 min, 55 degrees C for 10 min, and 60 degrees C for less than 1 min. The viability of spores was unaffected by ultrasonic irradiation for up to 4 min. Spores of G. caledonius and A. laevis were extremely resistant to ultraviolet radiation. Their viability was unaffected by exposure to 5 x 10 ergs cm from an ultraviolet source of 253.7nm. The spores had very thick, pigmented walls, and the possibility that these provided some protection against the physical and chemical treatments is discussed. The degree of physiological damage to the spores caused by the treatments demonstrated some adverse effects of basic laboratory procedures. This information, together with that on the comparative sensitivity of contaminating microbes to the treatments, was used in the development of protocol for producing large numbers of uncontaminated spores. PMID- 16345580 TI - New Selective Growth Inhibitors of Sphaerotilus natans, Anslimins A and B. AB - Selective growth inhibitors of Sphaerotilus natans were detected in the culture broths of several strains of Streptomyces. Anslimins A and B were isolated from the culture broth of Streptomyces oganonensis by adsorption on resinous adsorbent followed by elution with 50% aqueous acetone. The A and B components were then separated by cellulose powder chromatography. Physicochemical parameters of anslimins A and B revealed that these compounds are new peptide-containing structures. Anslimins A and B inhibited growth of S. natans at 0.78 and 0.39 mug/ml, respectively, but showed no activity against any of the other test microorganisms at 100 to 200 mg/ml. The anslimins had no harmful effect on guppies at a concentration of 100 mg/liter. PMID- 16345581 TI - Thiosulfate Oxidation and Tetrathionate Reduction by Intact Cells of Marine Pseudomonad Strain 16B. AB - Levels of thiosulfate-oxidizing enzyme (TSO) and tetrathionate reductase (TTR) were measured in washed cell suspensions of a heterotrophic marine thiosulfate oxidizing bacterium, strain 16B. TSO activity remained virtually constant in aerobically and anaerobically grown cells and was unaffected by the presence or absence of thiosulfate and tetrathionate in the growth medium. TTR was also present in cells grown aerobically and anaerobically, but its activity was threefold greater in cells cultured in media containing tetrathionate or thiosulfate. Tetrathionate appears to be the inducer of increased TTR activity in both aerobically and anaerobically grown cells. TTR (constitutive or induced) and TSO have different pH and temperature optima. Both TTR activities were unaffected by 10 mM KCN, which reversed oxygen inhibition of tetrathionate reduction. TSO was partially inhibited by 5 muM KCN and completely inhibited by 90 muM KCN. These findings and results of experiments to determine the influence of several inorganic electron donors and acceptors on TSO and TTR activities suggest that constitutive TSO and TTR represent reverse activities of the same enzyme, whereas inducible TTR is a separate enzyme used by strain 16B only for anaerobic respiration of tetrathionate. The bacterium appears well adapted to growth in environments characterized by low oxygen tension, dilute organic carbon concentrations, and the presence of a variety of reduced, inorganic sulfur compounds. PMID- 16345582 TI - Growth of Aeromonas hydrophila at Low Concentrations of Substrates Added to Tap Water. AB - The ability of an Aeromonas hydrophila isolate obtained from filtered river water to grow at low substrate concentrations was studied in batch experiments with tap water supplied with low concentrations of substrates. Growth was assessed by colony count determinations. The isolate only multiplied in the used tap water (2 to 3 mg of dissolved organic carbon per liter) after the addition of a small amount of an assimilable carbon compound. d-Glucose especially caused growth of the organism even at initial concentrations below 10 mug of C per liter. At initial glucose concentrations below the K(s) value (12 mug of C per liter), generation times and yield (colony-forming units per milligram of substrate-C) were nonlinear with 1/initial glucose concentrations and initial glucose concentrations, respectively. From these observations, the maintenance coefficient m was calculated (m = 0.015 mg of glucose per mg [dry wt] per h at 12 degrees C). At initial concentrations below the K(s) value of starch (73 mug of C per liter), no growth was observed, but complete use of starch occurred in these situations after the addition of 10 mug of glucose-C per liter. The results of this study show that information of ecological significance may be obtained by very simple batch experiments. Moreover, the isolate studied may be used in growth experiments to assess the maximum concentration of glucose which might be present in water, particularly tap water. PMID- 16345583 TI - Characterization of benthic microbial community structure by high-resolution gas chromatography of Fatty Acid methyl esters. AB - Fatty acids are a widely studied group of lipids of sufficient taxonomic diversity to be useful in defining microbial community structure. The extraordinary resolution of glass capillary gas-liquid chromatography can be utilized to separate and tentatively identify large numbers of fatty acid methyl esters derived from the lipids of estuarine detritus and marine benthic microbiota without the bias of selective methods requiring culture or recovery of the microbes. The gas-liquid chromatographic analyses are both reproducible and highly sensitive, and the recovery of fatty acids is quantitative. The analyses can be automated, and the diagnostic technique of mass spectral fragmentation analysis can be readily applied. Splitless injection on glass capillary gas chromatographic columns detected by mass spectral selective ion monitoring provides an ultrasensitive and definitive monitoring system. Reciprocal mixtures of bacteria and fungi, when extracted and analyzed, showed progressive changes of distinctive fatty acid methyl esters derived from the lipids. By manipulating the environment of an estuarine detrital microbial community with antibiotics and culture conditions, it was possible to produce a community greatly enriched in eucaryotic fungi, as evidenced by scanning electron microscopic morphology. The fatty acid methyl esters from the lipids in the fungus-enriched detritus showed enrichment of the C(18) dienoic and the C(18) and C(20) polyenoic esters. Manipulation of the detrital microbiota that increased the procaryotic population resulted in an absence of large structures typical of fungal mycelia or diatoms, as evidenced by scanning electron microscopy, and a significantly larger proportion of anteiso- and isobranched C(15) fatty acid esters, C(17) cyclopropane fatty acid esters, and the cis-vaccenic isomer of the C(18) monoenoic fatty acid esters. As determined by these techniques, a marine settling community showed greater differences in bacterial as contrasted to microeucaryotic populations when compared with the microbial communities of benthic cores. PMID- 16345584 TI - Isolation Method for Bacterial Isolates Capable of Growth on p-Chlorobiphenyl. AB - A method is reported for screening for p-chlorobiphenyl (pCB)-degrading bacteria from various environments. A solid medium was inoculated with the sample to be analyzed, colonies were allowed to develop, and the plates were then sprayed with a pCB solution in ether. The positive colonies were recorded as those surrounded with a clear zone in the film of pCB. That these colonies were able to degrade pCB was shown by their ability to grow on pCB in liquid medium with concomitant disappearance of the substrate and by the appearance of colored compounds in cultures grown on pCB. PMID- 16345586 TI - Simultaneous concentration of salmonella and enterovirus from surface water by using micro-fiber glass filters. AB - [This corrects the article on p. 659 in vol. 39.]. PMID- 16345585 TI - Utilization of dichloromethane by suspended and fixed-film bacteria. AB - Dichloromethane (methylene chloride) was biodegraded by and supported growth of suspended and fixed-film bacteria enriched from sewage. PMID- 16345587 TI - Uric Acid-Degrading Bacteria in Guts of Termites [Reticulitermes flavipes (Kollar)]. AB - Uricolytic bacteria were present in guts of Reticulitermes flavipes in populations up to 6 x 10 cells per gut. Of 82 strains isolated under strict anaerobic conditions, most were group N Streptococcus sp., Bacteroides termitidis, and Citrobacter sp. All isolates used uric acid (UA) as an energy source anaerobically, but not aerobically, and NH(3) was the major nitrogenous product of uricolysis. However, none of the isolates had an absolute requirement for UA. Utilization of heterocyclic compounds other than UA was limited. Fresh termite gut contents also degraded UA anaerobically, as measured by CO(2) evolution from [2-C]UA. The magnitude of anaerobic uricolysis [0.67 pmol of UA catabolized/(gut x h)] was entirely consistent with the population density of uricolytic bacteria in situ. Uricolytic gut bacteria may convert UA in situ to products usable by termites for carbon, nitrogen, energy, or all three. This possibility is consistent with the fact that R. flavipes termites from UA, but they do not void the purine in excreta despite the lack of uricase in their tissues. PMID- 16345588 TI - Anaerobic degradation of uric Acid by gut bacteria of termites. AB - A study was done of anaerobic degradation of uric acid (UA) by representative strains of uricolytic bacteria isolated from guts of Reticulitermes flavipes termites. Streptococcus strain UAD-1 degraded UA incompletely, secreting a fluorescent compound into the medium, unless formate (or a formicogenic compound) was present as a cosubstrate. Formate functioned as a reductant, and its oxidation to CO(2) by formate dehydrogenase provided 2H + 2e needed to drive uricolysis to completion. Uricolysis by Streptococcus UAD-1 thus corresponded to the following equation: 1UA + 1formate --> 4CO(2) + 1acetate + 4NH(3). Urea did not appear to be an intermediate in CO(2) and NH(3) formation during uricolysis by strain UAD-1. Formate dehydrogenase and uricolytic activities of strain UAD-1 were inducible by growth of cells on UA. Bacteroides termitidis strain UAD-50 degraded UA as follows: 1UA --> 3.5 CO(2) + 0.75acetate + 4NH(3). Exogenous formate was neither required for nor stimulatory to uricolysis by strain UAD-50. Studies of UA catabolism by Citrobacter strains were limited, because only small amounts of UA were metabolized by cells in liquid medium. Uricolytic activity of such bacteria in situ could be important to the carbon, nitrogen, and energy economy of R. flavipes. PMID- 16345589 TI - Enumeration of particle-bound and unattached respiring bacteria in the salt marsh environment. AB - Proportions of respiring bacteria determined with a 2-(p-iodophenyl)-3-(p nitrophenyl)-5-phenyl tetrazolium chloride dye-epifluorescent technique were significantly elevated in the 300-mum surface layer of a salt marsh estuary. Almost all the detectably respiring bacteria in the particle-laden surface layer and a significant proportion in subsurface waters were attached to particles. PMID- 16345590 TI - Incorporation of Specific Fatty Acid Precursors During Spore Germination and Outgrowth in Bacillus thuringiensis. AB - The selective incorporation of precursors specific for individual fatty acids in germinating and outgrowing spores of Bacillus thuringiensis is described. The specific precursors utilized were [C]butyrate, -isobutyrate, -valerate, and isovalerate, which were incorporated into even-numbered normal-chain isomers, even-numbered iso-isomers, odd-numbered normal-chain acids, and odd-numbered isohomologs, respectively. This preferential incorporation by B. thuringiensis allows the terminal carbons of specific normal and branched-chain fatty acids, contained within the cytoplasmic membrane, to be labeled with C and, potentially, C. PMID- 16345591 TI - Decomposition of Lignocellulose by Cyathus stercoreus (Schw.) de Toni NRRL 6473, a "White Rot" Fungus from Cattle Dung. AB - Cyathus stercoreus (Schw.) de Toni NRRL 6473, isolated from aged and fragmented cattle dung collected from a Michigan pasture, effected substantial losses in lignin (45%) from wheat straw during a 62-day fermentation (25 degrees C). The basidiomycete also improved wheat straw digestibility by freeing alpha-cellulose for enzymatic hydrolysis to glucose (230 mg of glucose per 1,000 mg of fermented residue). The rationale for selecting C. stercoreus in attempting to biologically modify the lignin and cellulose components in wheat straw or other gramineous agricultural residues was based on the expectation that this organism is ecologically specialized to enzymatically attack the substructures of native lignins in grasses. PMID- 16345592 TI - Inhibition of bacteria by lactulose preparations. AB - Lactulose syrups were similar to sucrose syrups in water activity-lowering effects but were more inhibitory toward test microorganisms. Heat-treated commercial lactulose syrups were most inhibitory, whereas non-heat-treated pure lactulose was only slightly more inhibitory than sucrose. PMID- 16345593 TI - Production of l-Glutamine by a Penicillin-Resistant Mutant of Flavobacterium rigense. AB - To establish a practical method for the fermentative production of l-glutamine, cultural conditions for the accumulation of a large amounts of l-glutamine were investigated by using Flavobacterium rigense 703, which was previously reported by us as a l-glutamine-producing mutant. As a result, a yield of 25 mg of l glutamine per ml was obtained after a 48-h cultivation in a medium containing glucose, yeast extract, (NH(4))(2)-fumarate, KH(2)PO(4), K(2)HPO(4), MgSO(4).7H(2)O, and CaCO(3) (pH 6.4). Accumulation of l-glutamine was dependent upon the concentration of (NH(4))(2)-fumarate, and a suboptimum growth at a relatively high concentration of (NH(4))(2)-fumarate was essential for the maximum production of l-glutamine. At the optimum conditions, glutamic acid was formed as a by-product at a concentration of less than 1 mg/ml, but accumulation of the other amino acids was negligible. The product was isolated from the culture broth and readily purified by anion-exchange chromatography. The pure crystals of l-glutamine obtained in an 80% yield were optically and chromatographically pure. PMID- 16345594 TI - Lignocellulose mineralization by arctic lake sediments in response to nutrient manipulation. AB - Mineralization of specifically labeled C-cellulose- and C-lignin-labeled lignocelluloses by Toolik Lake, Alaska, sediments was examined in response to manipulation of various environmental factors. Mineralization was measured by quantifying the amount of labeled CO(2) released from the specifically labeled substrates. Nitrogen (NH(4)NO(3)) and, to a greater degree, phosphorus (PO(4)) additions enhanced the mineralization of white pine (Pinus strobus) cellulose during the summer of 1978. Nitrogen and phosphorus together had no cumulative effect. During the summer of 1979, nitrogen or phosphorus alone had only a slight stimulatory effect on the mineralization of a sedge (Carex aquatilis) cellulose; however, together, they had a dramatic effect. This variable response of mineralization to nutrient addition between 1978 and 1979 was probably attributable to year-to-year variation in nutrient availability within the lake. Cellobiose addition and oxygen depletion inhibited the amount of pine cellulose mineralized. Whereas addition of nitrogen to oxygen-depleted treatments had limited effect, addition of phosphorus resulted in mineralizations equal to or greater than that of the controls. Nitrogen had no effect on mineralization of pine or Carex lignins. Phosphorus, however, inhibited mineralization of both lignins. With Carex lignin, the phosphorus inhibition occurred at a concentration as low as 0.1 muM. The antagonistic role of phosphorus in cellulose and lignin mineralizations may be of significance in understanding the increased proportion of lignin relative to cellulose in decomposing litter. PMID- 16345595 TI - Effects of Environmental Factors on Microbial Populations in Brackish Waters off the Southern Coast of Finland. AB - The roles played by environmental factors in seasonal changes in microbial populations were investigated in the Tvarminne area, off the southern coast of Finland. Surface-layer samples were collected at 1- or 2-week intervals in 1976 78, and 14 microbiological and 10 environmental parameters were determined. Stepwise multiple regression analysis was used to explain seasonal variation in the microbiological parameters. Separate analyses were made of the data from the open-water and ice-cover periods. In analyses of data from both periods, the environmental factors included accounted for a significant proportion of the variation in the parameters for community respiration (90%) and bacterial spores (80%), and a smaller proportion (60 to 65%) of the variation in total counts of bacteria and plate counts of psychrophiles and yeasts. Lower values (40 to 55%) were obtained for the variation in the other microbiological parameters. The environmental factors with maximal contributions were organic matter, water temperature, chlorophyll a, and salinity, but rainfall and winds also explained part of the variation in some microbiological parameters. In the winter analysis the results differed from those obtained for the other seasons, the variation being governed by parameters indicating freshwater outflows, namely, humic matter, salinity, water temperature (positive regression coefficient), and rainfall (negative regression coefficient). PMID- 16345596 TI - Factor analysis of the impact of the environment on microbial communities in the tvarminne area, southern coast of Finland. AB - Data already examined by regression analysis were subjected to factor analysis to scrutinize the effects of environmental factors on microbial populations in the brackish waters of the Tvarminne archipelago on the southern coast of Finland. Water samples were collected from 1.0-m depth at one point in Tvarminne Storfjard, 71 times over about 2 years. Twenty-six parameters were determined on each sample, 10 of environmental and 16 of microbiological type. The correlations between the parameters were factorized using the principal axis solution, and eight factors chosen for further consideration were rotated by the varimax method. The major part of the variance (about 90% of the total communality) of the microbiological parameters was covered by five factors, interpreted as phytoplankton blooms, the periods before and after the blooms, freshwater outflows, and water temperature. Wind variables were components in the factors interpreted as freshwater outflows. Rainfall played a minor part in the total variance of the microbial community, but it washed yeasts and proteolytic bacteria from the land into the study area. The eight factors selected covered about 60 to 98% of the variance of the microbiological parameters. The highest values (above 90%) were obtained for direct counts of bacteria, for plate counts of mesophilic and polymyxin-resistant bacteria, and for the two community respiration parameters; the lowest values (60 to 75%) were obtained for H(2)S producing and proteolytic bacteria. PMID- 16345597 TI - Terminal processes in the anaerobic degradation of an algal-bacterial mat in a high-sulfate hot spring. AB - The algal-bacterial mat of a high-sulfate hot spring (Bath Lake) provided an environment in which to compare terminal processes involved in anaerobic decomposition. Sulfate reduction was found to dominate methane production, as indicated by comparison of initial electron flow through the two processes, rapid conversion of [2-C]acetate to CO(2) and not to CH(4), and the lack of rapid reduction of NaHCO(3) to CH(4). Sulfate reduction was the dominant process at all depth intervals, but a marked decrease of sulfate reduction and sulfate-reducing bacteria was observed with depth. Concurrent methanogenesis was indicated by the presence of viable methanogenic bacteria and very low but detectable rates of methane production. A marked increased in methane production was observed after sulfate depletion despite high concentrations of sulfide (>1.25 mM), indicating that methanogenesis was not inhibited by sulfide in the natural environment. Although a sulfate minimum and sulfide maximum occurred in the region of maximal sulfate reduction, the absence of sulfate depletion in interstitial water suggests that methanogenesis is always severely limited in Bath Lake sediments. Low initial methanogenesis was not due to anaerobic methane oxidation. PMID- 16345598 TI - R-Plasmid Transfer in Zymomonas mobilis. AB - Conjugal transfer of three IncP1 plasmids and one IncFII plasmid into strains of the ethanol-producing bacterium Zymomonas mobilis was obtained. These plasmids were transferred at high frequencies from Escherichia coli and Pseudomonas aeruginosa into Z. mobilis and also between different Z. mobilis strains, using the membrane filter mating technique. Most of the plasmids were stably maintained in Z. mobilis, although there was some evidence of delayed marker expression. A low level of chromosomal gene transfer, mediated by plasmid R68.45, was detected between Z. mobilis strains. Genetic evidence suggesting that Z. mobilis may be more closely related to E. coli than to Pseudomonas or Rhizobium is discussed. PMID- 16345599 TI - Variation in heterotrophic and autotrophic nitrifier populations in relation to nitrification in organic soils. AB - The occurrence of heterotrophic and autotrophic nitrifiers in Pahokee muck and the role of these organisms in the ecosystem were assessed by surveying their population densities under different field conditions and by observing the relationship of these populations with aerobic bacteria and soil moisture. Heterotrophic nitrifier populations varied from 2.0 x 10 to 3.8 x 10 bacteria per cm of muck in surface fallow (bare) Pahokee muck during the annual cycle. This population decreased 40-fold between the surface and the 60- to 70-cm depths of soil. Similar variations were noted with autotrophic nitrifier populations. Significant correlations were found between heterotrophic nitrifiers and both soil moisture and aerobic bacteria. These relationships did not exist for the autotrophic nitrifiers. In soil that had been heated to kill the autotrophic nitrifiers, while preserving a population of the heterotrophs, and then amended with sodium acetate or ammonium sulfate or both, no nitrate or nitrite accumulated, although significant increases in heterotrophic nitrifiers were detected. In unheated control soil, nitrate plus nitrite-N increased from 14.3 to 181 mug/g of wet soil, and 48 mug of nitrite-N per g was produced. These data suggest that the autotrophic nitrifiers were the sole population responsible for nitrification in Pahokee muck. PMID- 16345600 TI - Comparison of bacterial extracellular polymer extraction methods. AB - Five different bacterial extracellular polymer extraction methods and a combination of two of these methods were compared on cultures of activated sludge, synthetic activated sludge, and Klebsiella aerogenes. High-speed centrifugation was the most effective extraction method for the K. aerogenes culture, based on the comparatively small amount of cell disruption and the relatively high extracellular polymer yield. Steaming treatment was the most effective extraction method for the activated sludges, since it released a significant quantity of extracellular polymers from the flocs and caused less cellular disruption than ethylenediaminetetraacetic acid and sodium hydroxide treatments. Sodium hydroxide treatment caused extensive disruption in all cultures. Ultrasonication released low concentrations of extracellular polymers from all cultures. However, it caused no significant cell disruption and therefore may be useful as a preliminary treatment in conjunction with another extraction method. PMID- 16345601 TI - Thermothrix thiopara: Growth and Metabolism of a Newly Isolated Thermophile Capable of Oxidizing Sulfur and Sulfur Compounds. AB - Thermothrix thiopara is isolated from hot sulfur springs. It occurs in situ at a temperature of 72 degrees C, a pH of 7.0, and an HS concentration of 17.4 mumol/liter (0.8 ppm). The organism was capable of autotrophic growth. Sulfite, sulfur, and polythionates were formed and subsequently degraded to sulfate during growth with thiosulfate as the sole energy source. Thiosulfate was oxidized by the polythionate pathway, and the stoichiometry of growth on thiosulfate was determined. The organism was also capable of heterotrophic growth in amino acids and simple sugars. A source of reduced sulfur (methionine, glutathione) was required for heterotrophic growth. Growth occurred aerobically or anaerobically with nitrate as a terminal oxidant. Both nitrous oxide and dinitrogen were produced. At 73 degrees C the maximum autotrophic growth rate in batch culture using thiosulfate was 0.56 generation per h. Under the same conditions in continuous culture, washout occurred at a dilution rate of 0.3 to 0.4 per h, corresponding to a cellular growth rate of 0.43 to 0.58 generation per h. This was nearly three times the growth rate for Thiobacillus denitrificans. T. thiopara is gram negative. It was also found to be both lysozyme and penicillin susceptible. As a result, this organism cannot be considered an archaebacterium. PMID- 16345602 TI - Effect of temperature and retention time on methane production from beef cattle waste. AB - The effect of temperature and retention time on the rate of methane production from waste of beef cattle fed a finishing diet was investigated by using continuously mixed 3-liter working volume anaerobic fermentors. The temperatures ranged from 30 to 65 degrees C with 5 degrees C increments between fermentors. The fermentors were fed once per day with 6% volatile solids (organic matter). Retention time for each temperature was varied from 18 to 2.5 days. After 3 volume turnovers, samples were obtained on 4 consecutive days. The highest methane production rate (liters/liter of fermentor per day) and methane yield at that rate (liters/gram of volatile solids) were 1.27 and 0.19 at 9 days and 30 degrees C, 1.60 and 0.16 at 6 days and 35 degrees C, 2.28 and 0.23 at 6 days and 40 degrees C, 2.42 and 0.24 at 6 days and 45 degrees C, 2.83 and 0.14 at 3 days and 50 degrees C, 2.75 and 0.14 at 3 days and 55 degrees C, 3.18 and 0.14 at 2.5 days and 60 degrees C, and 1.69 and 0.17 at 6 days and 65 degrees C. Volatile solids degradation at these retention times and temperatures was between 46 and 54%. The concentrations of volatile acids in the 30 to 55 degrees C fermentors were generally below 2,000 mg/liter, with the exception of the 3-day retention time. The 60 and 65 degrees C fermentors were usually above this level for all retention times. These studies indicate potential rates of methane production from the fermentation of untreated waste of beef cattle fed high-grain finishing diets. This information should serve as preliminary guidelines for various kinetic analyses and aid in economic evaluations of the potential feasibility of fermenting beef cattle waste to methane. PMID- 16345603 TI - Denitrification in Low pH Spodosols and Peats Determined with the Acetylene Inhibition Method. AB - Potential denitrification rates were determined for predominantly acid (pH >/= 3.6) horizons of forestal, miry, and agricultural soils from 22 locations in southern Finland. The acetylene inhibition method was used with nitrate-amended water-logged soils incubated in an N(2) atmosphere containing 2.5 or 5% C(2)H(2). Complete inhibition of the reduction of N(2)O to N(2) was observed in 99.3% of the samples. The denitrification rates varied from 0.12 to 53.8 mug of N.cm.day. Correlation between denitrification rate and soil pH was highly significant: r = 0.84 on a volume basis, and r = 0.44 on a weight basis. Vegetation type and amount of soil organic matter had a minor or no effect, respectively. In spodosolized soils the rates were significantly higher for B horizons than for A horizons. These results show that denitrification can occur in acid soils. PMID- 16345604 TI - Lipase Induction in Mucor hiemalis. AB - The influence on lipase induction in Mucor hiemalis of different types of triglycerides containing mainly oleic acid (olive oil), erucic acid (mustard oil), or saturated fatty acids of 8 to 16 carbons (coconut oil) was studied. The fungus was grown in shake flasks in a fermentation medium containing peptone, minerals, and glucose or one of the oils as the carbon source. Maximum lipase was produced when the initial pH of the fermentation medium was kept at 4.0. Addition of Ca to the medium did not increase lipase production. The optimum pH for activity of both the mycelial and extracellular lipases was found to be 7.0. The fungus produced a significant amount of lipase in the presence of glucose, but the lipase activity increased markedly when olive oil was added to the medium at the beginning of the fermentation. Addition of olive oil at a later stage did not induce as much enzyme. Studies with washed mycelia showed that a greater amount of lipase was released when olive oil was present than when glucose was present. Among the various types of triglycerides used as the carbon source, olive oil was found to be most effective in inducing the lipase. Olive oil and mustard oil fatty acids inhibited the lipase more than those of coconut oil. The lipase induced by a particular type of triglyceride did not seem to be specific for the same triglyceride, nor was it inhibited specifically by it. Irrespective of the triglyceride used in the fermentation medium, the lipase produced was most active against coconut oil triglyceride, and this specificity, as shown by lipase activities in an n-heptane system, was not found to be due to a better emulsification of this oil. The lipase of M. hiemalis can be considered to be both constitutive and inducible. PMID- 16345605 TI - Microbial degradation of a macrotetrolide miticide in soil. AB - Macrotetrolide, a miticide consisting of tetranactin, trinactin, and dinactin, was readily biodegradable and hence did not accumulate in soil. [U C]macrotetrolide was rapidly degraded via its constituent hydroxycarboxylic acids to carbon dioxide and water. In culture media, however, the mixture was hydrolyzed to homononactic and nonactic acids by three strains of Bacillus sp. and two of Micrococcus sp. The latter strains were able to hydrolyze 500 mug of the antibiotic per ml within a few days and to grow in the presence of 4,000 mug of the antibiotic per ml. However, they were unable to assimilate the constituent acids which accumulated in the culture medium. PMID- 16345606 TI - Improved filtration technique for concentrating and harvesting bacteria. AB - An improved technique is described for the filtrative concentration and harvesting of bacterial cultures. A pleated tangential flow filtration unit containing 1,000 cm of 0.2-mum-pore-size microporous membrane was used to rapidly (30 to 50 min) reduce the volume of 5 liters of bacterial culture of approximately 10 cells per ml to 0.2 to 0.5 liters of concentrated bacterial suspension. The effects of cell concentration, filtration pressure, and tangential flow rate were examined with respect to the rate of concentration and cell viability. Recovery efficiencies were between 60 and 75%, with no apparent impairment of organism viability. Cell concentration exerted the predominant effect on the filtration rate. PMID- 16345607 TI - Comparison of in situ and in vitro rates of methane release in freshwater sediments. AB - Anaerobic lake sediment incubated in vitro was investigated for its ability to mimic natural in situ sediment activities, using rate of methane production for the comparison. Two lakes with different rates and seasonal patterns of methanogenic activity were compared. There was good agreement (at the 97.5% confidence level) between rates of in situ methane release and initial (lasting an average of 120 h) rates of production measured in vitro in surface (0- to 3 cm) sediment. Evidence from this study, and others, indicated that it is the in situ surface sediment methane production which is primarily responsible for maintaining in situ methane release, and thus the above agreement was what was expected if surface in situ activity was maintained in vitro. When deeper sediment was investigated, however, the sum of in vitro rates from 0 to 20 cm (measured in 1.5- to 3-cm intervals) was much higher than in situ release rates and would have resulted in an impossibly high volume of gas. The extra gas could not have been stored within the sediments. We conclude that the in situ methanogenic activity of the 0- to 3-cm anaerobic surface sediments could be preserved during removal and laboratory incubation. However, similar treatment of deeper sediment appeared to stimulate methanogenic activity. PMID- 16345608 TI - Nutritional Interdependence Among Rumen Bacteria, Bacteroides amylophilus, Megasphaera elsdenii, and Ruminococcus albus. AB - Nutritional interdependence among three representatives of rumen bacteria, Bacteroides amylophilus, Megasphaera elsdenii, and Ruminococcus albus, was studied with a basal medium consisting of minerals, vitamins, cysteine hydrochloride, and NH(4). B. amylophilus grew well in the basal medium supplemented with starch and produced branched-chain amino acids after growth ceased. When cocultured with B. amylophilus in the basal medium supplemented with starch and glucose, amino acid-dependent M. elsdenii produced an appreciable amount of branched-chain fatty acids, which are essential growth factors for cellulolytic R. albus. A small addition of starch (0.1 to 0.3%) to the basal medium containing glucose and cellobiose brought about successive growth of the three species in the order of B. amylophilus, M. elsdenii, and R. albus, and successive growth was substantiated by the formation of branched-chain amino acids and fatty acids in the culture. Supplementation with 0.5% starch, however, failed to support the growth of R. albus. On the basis of these results, the effects of supplementary starch or branched-chain fatty acids on cellulose digestion in the rumen was discussed. PMID- 16345609 TI - Some effects of douglas fir terpenes on certain microorganisms. AB - The Douglas fir terpene alpha-pinene was shown to inhibit the growth of a variety of bacteria and a yeast. Other terpenes of the Douglas fir, including limonene, camphene, and isobornyl acetate, were also inhibitory to Bacillus thuringiensis. All terpenes were inhibitory at concentrations normally present in the fir needle diet of Douglas fir tussock moth larvae. The presence of such terpenes in the diet of these insects was found to strongly influence the infectivity of B. thuringiensis spores for the Douglas fir tussock moth larvae. The terpene alpha pinene destroyed the cellular integrity and modified mitochondrial activity in certain microorganisms. PMID- 16345610 TI - Variation in microbial activity in histosols and its relationship to soil moisture. AB - Microbial biomass, dehydrogenase activity, carbon metabolism, and aerobic bacterial populations were examined in cropped and fallow Pahokee muck (a lithic medisaprist) of the Florida Everglades. Dehydrogenase activity was two- to sevenfold greater in soil cropped to St. Augustinegrass (Stenotaphrum secundatum (Walt) Kuntz) compared with uncropped soil, whereas biomass ranged from equivalence in the two soils to a threefold stimulation in the cropped soil. Biomass in soil cropped to sugarcane (Saccharum spp. L) approximated that from the grass field, whereas dehydrogenase activities of the cane soil were nearly equivalent to those of the fallow soil. Microbial biomass, dehydrogenase activity, aerobic bacterial populations, and salicylate oxidation rates all correlated with soil moisture levels. These data indicate that within the moisture ranges detected in the surface soils, increased moisture stimulated microbial activity, whereas within the soil profile where moisture ranges reached saturation, increased moisture inhibited aerobic activities and stimulated anaerobic processes. PMID- 16345611 TI - Cage culture turbidostat: a device for rapid determination of algal growth rate. AB - The present cage culture turbidostat consists of a growth chamber and a control unit. The microorganisms (photoautotrophic algae) are kept in the growth chamber by porous membranes (pore size 1 to 3 mum) which retain the algae but allow efficient exchange of the growth medium. Flow rate and composition of the medium can therefore be varied independently of algal population density. A reciprocating pumping mode of the medium is introduced to obtain more gentle clearance of membranes than that provided by rotation or stirring in other membrane fermentors. Pulsed light and a light-emitting diode/light-sensitive transistor couple are used to monitor the turbidity of the culture, independent of external light needed for growth. The control unit keeps the turbidity constant by frequent activation of the dilution pump. Theoretical analysis of growth in the turbidostat shows that integrated activation time of the dilution pump is proportional to the growth rate of the organism. Theoretical analysis was also used to determine minimum flow-rate and nutrient concentration of medium to cover the requirement of the algae. Experiments with three different marine diatoms were carried out, and they demonstrated that the growth rate could be determined every hour and that the cultures could be kept at constant turbidity over 10 to 14 days at least. PMID- 16345613 TI - Short-term effects of South louisiana and kuwait crude oils on glucose utilization by marine bacterial populations. AB - Two crude oils, South Louisiana and Kuwait, were examined for their impact on glucose utilization by bacterial populations from the Gulf of Mexico. The uptake and mineralization of [U-C]glucose was assayed after a 4- to 23-h exposure to various concentrations of added crude oil (0, 0.001, 0.01, and 0.1% [vol/vol]). The effects of oil were determined in a total of 15 sediment and 13 water samples collected from offshore, open-bay, and salt marsh environments. The utilization of glucose by bacterial populations usually was not affected by added oil; in 10 sediment and 11 water samples, oil had no significant effect on either glucose uptake or mineralization. Stimulation by oil was recorded in four sediment samples. Oil inhibition occurred in one sediment and two water samples, but only in the presence of the highest concentration of added oil, i.e., 0.1%. Our data suggest that short-term exposure to either South Louisiana or Kuwait crude oil, even at 0.1%, usually has no toxic effect on glucose utilization by marine bacterial populations. PMID- 16345612 TI - Alteration of Host Specificity to Lytic Bacteriophages in Streptococcus cremoris. AB - A mutant of Streptococcus cremoris strain ML1 was isolated based on its resistance to acriflavine. The mutant strain showed resistance to the growth of virulent bacteriophages to which the parental strain was sensitive whereas it became sensitive to a number of other virulent phages to which the parental strain was resistant. At the same time, infection of the mutant strain by another bacteriophage sc607 resulted in killing of cells without production of progeny phages. The phage adsorption appeared normal, suggesting that the killing was a postadsorption event. Such killing of bacterial cells was prevented by chloramphenicol treatment, indicating that involvement of some protein either synthesized by phage or phage-induced cellular protein. Synthesis of ribonucleic acid was abruptly terminated after infection of the mutant strain by phage sc607 but not of the parental strain. The alteration of host specificity in the mutant to different lytic bacteriophages and especially abortive infection by phage sc607 resembles the prophage-mediated interference observed in other bacteria. PMID- 16345614 TI - Effect of Estuarine Sediment pH and Oxidation-Reduction Potential on Microbial Hydrocarbon Degradation. AB - Microbial mineralization rates of two petroleum hydrocarbons, as affected by pH and oxidation-reduction potential, were determined in a Barataria Bay, Louisiana, sediment using C-labeled hydrocarbons. Hydrocarbon mineralization rates were inferred from the activity of respired CO(2). Sediment pH and oxidation-reduction potential were important factors in governing the population of hydrocarbon degrading microorganisms in the sediment and subsequent mineralization rates. Highest mineralization rates occurred at pH 8.0, and the lowest occurred at pH 5.0. At all pH levels mineralization decreased with decreasing oxidation reduction potential (i.e., increasing sediment anaerobiosis). Generally, mineralization rates for octadecane were greater than those for naphthalene. Aerobic microorganisms in the oxidized sediment were more capable of degrading hydrocarbons than anaerobic microorganisms in reduced sediment of the same pH. PMID- 16345615 TI - Regulation of enzymes associated with C-1 metabolism in three facultative methylotrophs. AB - The levels of the oxidation enzyme methanol dehydrogenase and the serine pathway enzymes, hydroxypyruvate reductase, glycerate kinase, serine transhydroxymethylase, serine-glyoxylate aminotransferase, phosphoenolpyruvate carboxylase, and malyl-coenzyme A lyase, were studied in cells of the facultative methylotrophs Pseudomonas AM1, Pseudomonas 3A2 and Hyphomicrobium X grown on different substrates. Induction and dilution curves for these enzymes suggest they may be regulated coordinately in Hyphomicrobium X, but not in Pseudomonas AM1 or 3A2. Glyoxylate stimulated the serine transhydroxymethylase activity in methanol-grown cells of all three organisms. A secondary alcohol dehydrogenase activity was detected at low levels in Pseudomonas AM1 and Hyphomicrobium X, but not in Pseudomonas 3A2. PMID- 16345616 TI - Degradation of C-Labeled Lignins and C-Labeled Aromatic Acids by Fusarium solani. AB - Abilities of isolate AF-W1 of Fusarium solani to degrade the side chain and the ring structure of synthetic dehydrogenative polymerizates, aromatic acids, or lignin in sound wood were investigated under several conditions of growth substrate or basal medium and pH. Significant transformations of lignins occurred in 50 days in both unextracted and extracted sound wood substrates with 3% malt as the growth substrate and the pH buffered initially at 4.0 with 2,2 dimethylsuccinate. Degradation of lignin in such woods also occurred under unbuffered pH conditions when a basal medium of either 3% malt or powdered cellulose in deionized water was present. Decomposition of the lignin in these woods did not occur in cultures where d-glucose was present as a growth substrate. F. solani significantly transformed, as measured as evolved CO(2), both synthetic side chain (beta, gamma)-C- and U-ring-C-labeled lignins in 30 days under liquid culture conditions of only distilled deionized water and no pH adjustment. Degradation of dehydrogenative polymerizates by F. solani was reduced drastically when D(2) was the liquid medium. AF-W1 also cleaved the alpha-C from p-hydroxybenzoic acid and evolved CO(2) from the substrate, [3-C]cinnamic acid. Thus, the fungus cleaved side chain carbon from substrate that originally lacked hydroxyl substitution on the aromatic nucleus. Surprisingly, small amounts of C cleaved from aromatic acids by F. solani were incorporated into cell mass. Initial buffering of the culture medium to pH 4.0 or 5.0 with 0.1 M 2,2 dimethylsuccinate significantly increased F. solani degradation of all lignins or aromatic acids. Results indicated that AF-W1 used lignin as a sole carbon source. PMID- 16345617 TI - Characterization of two new facultative methanotrophs. AB - Two new facultative methane-oxidizing bacteria have been isolated from lake water enrichments. The organisms have been characterized in terms of colony types, growth characteristics, the guanine plus cytosine content of their deoxyribonucleic acid, thin sections, oxidation rates, and carbon assimilation pathways. Methane-grown cells of both organisms contained intracytoplasmic membranes similar to those described as type II in other methanotrophic bacteria. Neither organism had such membranes when grown heterotrophically. Both organisms assimilated methane by way of the isocitrate lyase-negative serine pathway for formaldehyde incorporation. The enzymes of this pathway were high in specific activity in cells grown on methane and were at low levels in cells grown either on heterotrophic substrates or on heterotrophic substrates plus methane. It is proposed that both organisms be classified in the genus Methylobacterium as two new species, Methylobacterium ethanolicum and Methylobacterium hypolimneticum. PMID- 16345618 TI - Bacteria Associated with the Gut Tract of Larval Stages of the Aquatic Cranefly Tipula abdominalis (Diptera; Tipulidae). AB - Scanning electron microscopy, light microscopy, and direct isolations were used to examine the distribution and diversity of bacteria in the gut tracts of larval stages of Tipula abdominalis. The animal had an enlarged hindgut which housed a diverse bacterial community in the lumen and directly attached to the gut wall. Distinct localization was noted, with the most dense and most diverse community anterior to the rectum. A distinct architecture of bacteria occurred in this region, characterized by a layering or a "weblike" array of filamentous bacteria overlying mats of bacteria closely associated with the gut wall. Although morphological diversity was high in the hindgut, filamentous bacteria were the dominant morphology observed. The attached microbiota, sloughed during ecdysis, recolonized to the same density and diversity observed before the molt. The majority of the isolatable bacterial types were facultatively anaerobic. The distinct localization and attached nature of the hindgut bacteria and the recolonization after each molt suggest they are indigenous to this region of the gut tract. PMID- 16345619 TI - Molds in brined cucumbers: cause of softening during air-purging of fermentations. AB - Softening of cucumbers in fermentations purged at high air-flow rates was caused by molds growing in the brined cucumbers, not in the brine. This conclusion is based on the following results: (i) no microorganisms were isolated in significant numbers from brines that caused softening of pasteurized brined cucumbers, (ii) no pectinolytic enzyme activities were produced in cucumber brines in the absence of cucumbers, (iii) the pickles in some air-purged fermentations became very soft without the appearance of any pectinolytic enzyme activity in the brine, (iv) mold hyphae were consistently observed in tissues of soft pickles, (v) molds consistently developed in cultures of slices of surface sterilized cucumbers taken from fermentations in which soft pickles were subsequently found, and (vi) molds belonging to the genera Alternaria, Fusarium, and Mucor isolated from slices all softened pasteurized brined cucumbers. PMID- 16345620 TI - Nitrate dissimilation under microaerophilic conditions by a magnetic spirillum. AB - During microaerophilic growth of magnetic spirillum MS-1 on tartrate and nitrate, a maximal cell density was obtained at an initial oxygen partial pressure of 17 Pa. A transient accumulation of nitrous oxide and a 1:2 (mol/mol) stoichiometry between tartrate oxidation and nitrate reduction were observed, indicating that the organism carried out a respiratory type of metabolism. PMID- 16345622 TI - Die-away kinetics of aerosolized bacteria from sprinkler application of wastewater. AB - [This corrects the article on p. 1193 in vol. 39.]. PMID- 16345621 TI - Acetate as sole carbon and energy source for growth of methanosarcina strain 227. AB - [This corrects the article on p. 995 in vol. 39.][This corrects the article on p. 996 in vol. 39.]. PMID- 16345623 TI - Inhibition of the Anaerobic Growth of Brochothrix thermosphacta by Lactic Acid. AB - Brochothrix thermosphacta can grow aerobically in the presence of 210 mM l lactate and anaerobically in its absence at pH values down to at least 5.5. Anaerobic growth is, however, inhibited by l-lactate, the concentration of undissociated lactic acid being the governing factor. Postrigor meat usually contains sufficient lactic acid to select against the anaerobic growth of B. thermosphacta. At least some Lactobacillaceae strains are more resistant to lactic acid and so their growth is favored on vacuum-packaged meat. PMID- 16345624 TI - Role of microorganisms in the consumption and production of atmospheric carbon monoxide by soil. AB - Consumption and production of atmospheric CO was measured under field conditions in three different types of soil. CO was consumed by an apparent first-order reaction and produced by an apparent zero-order reaction, resulting in a dynamic equilibrium with the consumption of atmospheric CO as the net reaction. CO consumption was higher in summer than in winter. Laboratory experiments on five different soil types showed that CO consumption was strongly inhibited by the presence of streptomycin or cycloheximide (Actidione), or both. Thus, eucaryotic as well as procaryotic microorganisms were apparently responsible for the observed CO consumption. The aerobic carboxydobacterium Pseudomonas carboxydovorans added to sterile soil was able to utilize the low amounts (ca. 0.7 ppmv) of CO present in laboratory air. CO was consumed by soil under aerobic as well as anaerobic conditions. Anaerobic preincubation of the soil stimulated the anaerobic CO consumption and reduced the aerobic CO consumption. In contrast to CO consumption, CO production was stimulated by autoclaving, by ultraviolet irradiation, by fumigation with NH(3) or CHCl(3), by treatment with streptomycin or cycloheximide or both, by addition of NaCN, NaN(3), or Na(2)HAsO(4) (or all three) in the presence of glucose under an atmosphere of pure oxygen, or by a drying and rewetting procedure. The consumption of atmospheric CO by soil is a microbial process, but the production of CO is apparently not a metabolic process. PMID- 16345626 TI - Ethanol from Sugar Cane: Flask Experiments Using the EX-FERM Technique. AB - Alcohol production at the laboratory scale from sugar cane pieces by the EX-FERM technique was studied with 37 strains of Saccharomyces spp. The EX-FERM process is novel in that it employs the simultaneous extraction and fermentation of the sucrose in a cane-water suspension. Two types of cane treatments were used: chips and shredded pith, either fresh or dried. A mother culture of the yeast was prepared in enriched cane juice and then added to the cane-water mixture. After static fermentation for 40 h at 30 degrees C, the cane was removed, and fresh cane was added to the yeast-alcohol broth. After an additional 24 h, the cane was again removed and the liquor was analyzed. After the first 40-h cycle, sugar consumption was above 99% with 10 of the 37 yeast strains tested, and ethanol reached levels of 1.29 to 4.00 g per 100 ml, depending on the yeast strain. The final ethanol concentration reached 4.27 to 5.37 g per 100 ml, and sugar consumption was above 98% in three cases during a second EX-FERM cycle employing previously air-dried chips and pith. Product yields were within accepted values. Cane treatment did not appear to affect the results at this level. PMID- 16345625 TI - Antagonism between osmophilic lactic Acid bacteria and yeasts in brine fermentation of soy sauce. AB - Brine fermentation by osmophilic lactic acid bacteria and yeasts for long periods of time is essential to produce a good quality of shoyu (Japanese fermented soy sauce). It is well known that lactic acid fermentation by osmophilic lactic acid bacteria results in the depression of alcoholic fermentation by osmophilic yeasts, but the nature of the interaction between osmophilic lactic acid bacteria and yeasts in brine fermentation of shoyu has not been revealed. The inhibitory effect of osmophilic lactic acid bacteria on the growth of osmophilic yeasts was investigated. It was recognized that osmophilic shoyu yeasts such as Saccharomyces rouxii and Torulopsis versatilis were inhibited by a metabolite produced by osmophilic lactic acid bacteria (belonging to Pediococcus halophilus) in brine fermentation of shoyu. The primary inhibitor was considered to be acetic acid, although lactic acid was slightly inhibitory. PMID- 16345627 TI - Characteristics of some lactic Acid bacteria used as starter cultures in dry sausage production. AB - The fermentation characteristics of two commercial (Duploferment 66 and Saga II) and five Norwegian lactic acid bacteria used in dry sausage production were compared with those of Lactobacillus plantarum ATCC 8014. The Norwegian strains lacked the ability to ferment mannitol, sorbitol, lactose, and d-(+)-raffinose and grew at 8 but not at 42 degrees C, in contrast to the ATCC culture and the two commercial strains. The lactate dehydrogenase activity of the Norwegian strains was not stimulated by pyruvate. All strains were capable of peroxide destruction when grown in the presence of myoglobin. PMID- 16345628 TI - Evidence Suggesting Protozoan Predation on Rhizobium Associated with Germinating Seeds and in the Rhizosphere of Beans (Phaseolus vulgaris L.). AB - Changes in populations of microorganisms around germinating bean (Phaseolus vulgaris L.) seeds, in the rhizosphere of bean, and in a model rhizosphere were studied. Strains of Rhizobium phaseoli that were resistant to streptomycin and thiram were used, and as few as 300 R. phaseoli cells per g of soil could be enumerated with a selective medium that was devised. A direct role was not evident for bacterial competitors, lytic bacteria, antibiotic-producing microorganisms, bacteriophages, and Bdellovibrio in the suppression of R. phaseoli around germinating seeds and in the rhizosphere. Protozoa increased in numbers in the soil upon planting of the seeds. The extent of colonization of soil by R. phaseoli was inversely related to the presence of large numbers of bacteria and protozoa. Colonization of R. phaseoli was improved upon suppression of protozoa with thiram and also when the soil was amended with other protozoan inhibitors and mannitol to simulate seed and root exudation. The data support the view that the decrease in numbers of R. phaseoli is caused by an increase in protozoan predation, the protozoa increasing in number because they prey on bacteria that proliferate by using seed and root exudates as nutrients. PMID- 16345629 TI - Restriction and modification in group N streptococci: effect of heat on development of modified lytic bacteriophage. AB - The appearance of lytic bacteriophage against newly introduced starter strains used during commercial cheese manufacture occurs rapidly, and their origin is not well understood. In this study, members of the group N streptococci were examined for the presence of bacteriophage restriction and modification systems. Two streptococcal phages from Streptococcus cremoris TR and Streptococcus lactis C2 (phage designations tr and c2) showed restricted lytic development on S. cremoris 799 and KH, respectively. Efficiency of plaquing was 1.9 x 10 for tr plaqued on 799 and 2.1 x 10 for c2 plaqued on KH. After passage through the restrictive hosts, these phages demonstrated high lytic ability for formerly restrictive hosts. Stress of the restrictive host strains at temperatures of 40 to 50 degrees C resulted in a significant increase in the efficiency of plaquing of restricted bacteriophages. Elevated temperatures are encountered during commercial cheese manufacture. The results suggested that the temporary loss of host restriction activity with the resulting modification of nonspecific bacteriophage may contribute directly to the appearance of lytic phage against new starter strains. PMID- 16345630 TI - Effect of rice plants on nitrogenase activity of flooded soils. AB - In samples of flooded soil containing blue-green algae (cyanobacteria), the presence of rice plants did not influence the nitrogenase activity of the algae. Nitrogenase activity of heterotrophic bacteria was enhanced by the presence of rice plants, but this activity was not affected by changes in plant density. The rate of nitrogen fixation in the rhizosphere, however, varied significantly among the 16 rice varieties tested. A simple method was devised to test the nitrogen fixing activity in the root zone of rice varieties, and data were obtained showing marked differences in the activities of the 16 varieties. In tests of two varieties with dissimilar rates of nitrogen fixation in their rhizospheres, the variety which had the greater root weight and lesser shoot weight and which supported greater methane formation had the greater nitrogenase activity. PMID- 16345631 TI - Organic carbon utilization by resting cells of thiosulfate-oxidizing marine heterotrophs. AB - Two thiosulfate-oxidizing marine heterotrophs, strains 12W and 16B, were tested for utilization of [C]glucose and [C]acetate, respectively, in the presence or absence of thiosulfate. Thiosulfate oxidation caused an increase in organic carbon incorporation and a corresponding decrease in respiration at pH 6.5, near the optimum pH for thiosulfate oxidation and thiosulfate-stimulated growth in these bacteria. The amount of glucose or acetate metabolized remained virtually unaffected by thiosulfate oxidation. The metabolic shift in carbon utilization was diminished by increasing the initial pH to 8.0. The results indicate that marine heterotrophs 12W and 16B exhibit a type of mixotrophic metabolism which differs from that observed in the thiobacilli. PMID- 16345632 TI - Production of NO(2) and N(2)O by Nitrifying Bacteria at Reduced Concentrations of Oxygen. AB - Pure cultures of the marine ammonium-oxidizing bacterium Nitrosomonas sp. were grown in the laboratory at oxygen partial pressures between 0.005 and 0.2 atm (0.18 to 7 mg/liter). Low oxygen conditions induced a marked decrease in the rate for production of NO(2), from 3.6 x 10 to 0.5 x 10 mmol of NO(2) per cell per day. In contrast, evolution of N(2)O increased from 1 x 10 to 4.3 x 10 mmol of N per cell per day. The yield of N(2)O relative to NO(2) increased from 0.3% to nearly 10% (moles of N in N(2)O per mole of NO(2)) as the oxygen level was reduced, although bacterial growth rates changed by less than 30%. Nitrifying bacteria from the genera Nitrosomonas, Nitrosolobus, Nitrosospira, and Nitrosococcus exhibited similar yields of N(2)O at atmospheric oxygen levels. Nitrite-oxidizing bacteria (Nitrobacter sp.) and the dinoflagellate Exuviaella sp. did not produce detectable quantities of N(2)O during growth. The results support the view that nitrification is an important source of N(2)O in the environment. PMID- 16345633 TI - Fluorometric determination of adenosine nucleotide derivatives as measures of the microfouling, detrital, and sedimentary microbial biomass and physiological status. AB - Adenosine, adenine, cyclic adenosine monophosphate (AMP), AMP, nicotinamide adenine dinucleotide, adenosine diphosphate, and adenosine triphosphate (ATP) were recovered quantitatively from aqueous portions of lipid extracts of microfouling, detrital, and sedimentary microbial communities. These could be detected quantitatively in the picomolar range by forming their 1-N-etheno derivatives and analyzing by high-pressure liquid chromatography with fluorescence detection. Lipid extraction and subsequent analysis allowed the simultaneous measurement of the microbial community structure, total microbial biomass with the quantitative recovery of the adenine-containing cellular components, which were protected from enzymatic destruction. This extraction and fluorescent derivatization method showed equivalency with the luciferin luciferase method for bacterial ATP measurements. Quick-freezing samples in the field with dry ice-acetone preserved the ATP and energy charge (a ratio of adenosine nucleotides) for analysis at remote laboratories. The metabolic lability of ATP in estuarine detrital and microfouling communities, as well as bacterial monocultures of constant biomass, showed ATP to be a precarious measure of biomass under some conditions. Combinations of adenosine and adenine nucleotides gave better correlations with microbial biomass measured as extractable lipid phosphate in the detrital and microfouling microbial communities than did ATP alone. Stresses such as anoxia or filtration are reflected in the rapid accumulation of intracellular adenosine and the excretion of adenosine and AMP into the surrounding milieu. Increases in AMP and adenosine may prove to be more sensitive indicators of metabolic status than the energy charge. PMID- 16345634 TI - Effects of alkaline phosphatase activity on nucleotide measurements in aquatic microbial communities. AB - Alkaline phosphatase (APase) activity was detected in aquatic microbial assemblages from the subtropics to Antarctica. The occurrence of APase in environmental nucleotide extracts was shown to significantly affect the measured concentrations of cellular nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, guanosine triphosphate, uridine triphosphate, and cytidine triphosphate), adenylate energy charge, and guanosine triphosphate/adenosine triphosphate ratios, when conventional methods of nucleotide extraction were employed. Under the reaction conditions specified in this report, the initial rate of hydrolysis of adenosine triphosphate was directly proportional to the activity of APase in the sample extracts and consequently can be used as a sensitive measure of APase activity. A method was devised for obtaining reliable nucleotide measurements in naturally occurring microbial populations containing elevated levels of APase activity. The metabolic significance of APase activity in microbial cells is discussed, and it is concluded that the occurrence and regulation of APase in nature is dependent upon microscale inorganic phosphate limitation of the autochthonous microbial communities. PMID- 16345635 TI - Collagenase production by nematode-trapping fungi. AB - A number of species of nematode-trapping fungi, which capture and digest nematodes having keratin and collagen in their cuticles, were tested for the ability to produce extracellular collagenase and keratinase. Collagenase, which is active on ichthyocol, earthworm collagen, and procollagen from chicken embryo fibroblasts, was found in the growth medium of all tested species; keratinase was not found. The enzyme from Arthrobotrys amerospora was concentrated by precipitation with (NH(4))(2)SO(4) and further purified by adsorption on collagen at 0 degrees C. The collagenase was active over a pH range of 2.5 to 10.0. It was not inactivated by dialysis against ethylenediaminetetraacetic acid for 48 h or by the sulfhydryl group inhibitors N-ethylmaleimide and p-chloromercuribenzoate. The production of collagenase may aid the fungus to penetrate the cuticle of its prey. PMID- 16345636 TI - Aspects of diversity measurement for microbial communities. AB - A useful measure of diversity was calculated for microbial communities collected from lake water and sediment samples using the Shannon index (H') and rarefaction [E(S)]. Isolates were clustered by a numerical taxonomy approach in which limited (<20) tests were used so that the groups obtained represented a level of resolution other than species. The numerical value of diversity for each sample was affected by the number of tests used; however, the relative diversity compared among several sampling locations was the same whether 11 or 19 characters were examined. The number of isolates (i.e., sample size) strongly influenced the value of H' so that unequal sized samples could not be compared. Rarefaction accounts for differences in sample size inherently so that such comparisons are made simple. Due to the type of sampling carried out by microbiologists, H' is estimated and not determined and therefore requires a statement of error associated with it. Failure to report error provided potentially misleading results. Calculation of the variance of H' is not a simple matter and may be impossible when handling a large number of samples. With rarefaction, the variance of E(S) is readily determined, facilitating the comparison of many samples. PMID- 16345637 TI - Physiological adaptations in response to environmental stress during an n(2) fixing anabaena bloom. AB - Anabaena spiroides has the ability to maintain intense biomass production for extensive periods in the epilimnion of a small eutrophic lake characterized by conditions shown to cause photooxidative death in a number of other phytoplankton. By the enhancement of carotenoid synthesis chlorophyll a was protected from photooxidation and prevented from catalyzing other photooxidative reactions within the cells. By temporally separating CO(2) and N(2) fixation, maximum utilization of photosynthetically active radiation was achieved. Because CO(2) fixation was more sensitive than N(2) fixation to a high oxygen concentration, the former was maximized during morning hours, before the afternoon buildup of dissolved oxygen. The diurnal partitioning of carbon and N(2) fixation has two additional advantages; possible competition for reductant generating compounds is minimized, and adequate endogenous pools of carbon skeletons are assured to accept newly fixed ammonia. Hence, Anabaena, far from undergoing photooxidative death, appears to utilize a physiological strategy which allows optimization of radiant energy use for reductive processes and dominance of surface waters and shading of deeper phytoplankton during summer blooms. PMID- 16345638 TI - Fine Structure of Extracellular Polysaccharide of Erwinia amylovora. AB - Virulent E(9) and avirulent E(8) strains of Erwinia amylovora were shown by means of light, transmission, and scanning microscopy to be, respectively, encapsulated and unencapsulated. Difficulty was encountered in stabilizing the fibrillar appearing capsular extracellular polysaccharide. We suggest that the ephemeral nature of extracellular polysaccharide is due to the collapse of its extended structure upon dehydration. This occurs when bacteria are prepared for either transmission or scanning electron microscopy. The electron micrographs support our previous biochemical and immunological studies contending that the capsule is composed of tightly bound and loosely held components. The preparation of bacteria in freeze-dried colonies has permitted us to observe and explain the fluidity of the encapsulated strain. We suggest that this fluidity is a reflection of the loosely held extracellular polysaccharide or slime. PMID- 16345639 TI - Sequence of events in the digestion of fresh legume leaves by rumen bacteria. AB - When fresh whole leaves of six different species of forage legumes were suspended in an artificial rumen medium and inoculated with rumen bacteria, bacterial adhesion and proliferation were noted at the stomata, and penetration of the stomate by these bacteria was documented by electron microscopy. The invading bacteria adhered to surfaces within the intercellular space of the leaf and produced very extensive exopolysaccharide-enclosed microcolonies. After some of the legume leaf cell walls were disorganized and ruptured by bacterial digestion, these cells (notably, parenchyma and epidermal cells) were invaded by bacteria, with subsequent formation of intracellular microcolonies. However, other cells were neither ruptured nor colonized (notably, stomata guard cells and vascular tissue). At all stages of the digestion of intact legume leaves, the rumen bacteria grew in microcolonies composed of cells of single or mixed morphological types, and a particular ecological niche was often completely and consistently occupied by a very large microcolony of cells of single or mixed morphological types. PMID- 16345640 TI - Propionate-Degrading Bacterium, Syntrophobacter wolinii sp. nov. gen. nov., from Methanogenic Ecosystems. AB - A new genus and species of a nonmotile gram-negative rod, Syntrophobacter wolinii, is the first bacterium described which degrades propionate only in coculture with an H(2)-using organism and in the absence of light or exogenous electron acceptors such as O(2), sulfate, or nitrate. It was isolated from methanogenic enrichments from an anaerobic municipal sewage digestor, using anaerobic roll tubes containing a medium with propionate as the energy source in association with an H(2)-using, sulfate-reducing Desulfovibrio sp. which cannot utilize fatty acids other than formate. S. wolinii produced acetate and, presumably, CO(2) and H(2) (or formate) from propionate. In media without sulfate and with Methanospirillum hungatei, a methanogen that uses only H(2)-CO(2) or formate as an energy source, acetate, methane, and, presumably, CO(2) were produced from propionate and only small amounts of Desulfovibrio sp. were present. Isolation in coculture with the methanogen was not successful. S. wolinii does not use other saturated fatty acids as energy sources. PMID- 16345641 TI - Effects of grazing by estuarine gammaridean amphipods on the microbiota of allochthonous detritus. AB - Estuarine gammaridean amphipods grazing at natural population density on detrital microbiota affected the microbial community composition, biomass, and metabolic activity without affecting the physical structure of the leaves. Total microbial biomass estimated by adenosine triphosphate and lipid phosphate or observed by scanning electron microscopy was greater on grazed than on ungrazed detritus. The rates of oxygen consumption, poly-beta-hydroxybutyrate synthesis, total lipid biosynthesis, and release of CO(2) from radioactively prelabeled microbiota were higher on grazed than on ungrazed leaves, indicating stimulation of the metabolic activity of grazed detrital microbes. This was true with rates based either on the dry leaf weight or microbial biomass. Alkaline phosphatase activity was lower in the grazed system, consistent with enhanced inorganic phosphate cycling. The loss of C from both total lipid and poly-beta-hydroxybutyrate of microorganisms prelabeled with C was greater from grazed than ungrazed microbes. There was a faster decrease in the C-glycolipid than in the C-neutral lipid or C-phospholipid fractions. Analysis of specific phospholipids showed losses of the metabolically stable [C]glycerolphosphorylcholine derived from phosphatidylcholine and much more rapid metabolism of the bacterial lipid phosphatidylglycerol measured as [C]glycerolphosphorylglycerol with amphipod grazing. The biochemical data supported scanning electron microscopy observations of a shift as the grazing proceeded from a bacterial/fungal community to one dominated by bacteria. PMID- 16345642 TI - Enrichment of Auxotrophic Mutants in Streptomyces griseus. AB - A method for the isolation and enrichment of tryptophan auxotrophic mutants of the indolmycin-producing strain Streptomyces griseus ATCC 12648 was developed, using penicillin selection. With UV irradiation of 2.2 x 10 muW cm, a mutation rate to tryptophan auxotrophy of 5.6 x 10 was achieved. With 300 mug ml of penicillin G, an enrichment of tryptophan auxotrophs of about 1,000-fold was attained. Approximately 40% of all survivors were tryptophan auxotrophs. PMID- 16345643 TI - Physiological Changes Within a Metalimnetic Layer of Oscillatoria rubescens. AB - Different depths within a metalimnetic layer of Oscillatoria rubescens were sampled, and changes in biochemical composition and photosynthetic rate were analyzed. Although differences in pigment and protein content were found, the photosynthetic rates of the samples were not significantly different. PMID- 16345644 TI - Response of n(2)-fixing cyanobacteria to salt. AB - The effect of salt on photosynthetic activity, acetylene reduction, and related activities was examined in two species of cyanobacteria, Nostoc muscorum and Calothrix scopulorum. Photosynthesis was more resistant to high salt concentration than was N(2) fixation. The salt resistance of both activities increased after a period of exposure of the cells to salinity. The transfer of electrons via ferredoxin and ferredoxin-nicotinamide adenine dinucleotide phosphate reductase was found to be extremely sensitive to salt. In comparison, the transfer of reducing power by glucose-6-phosphate dehydrogenase, isocitric dehydrogenase, and photosystem 1 was less affected by NaCl, whereas glutamine synthetase exhibited higher tolerance to salt. PMID- 16345645 TI - Influence of cadmium, zinc, and lead on growth, trap formation, and collagenase activity of nematode-trapping fungi. AB - Growth and morphogenesis of seven species of nematode-trapping fungi and the activity of a collagenase produced by Arthrobotrys amerospora were measured in the presence of various concentrations of divalent cadmium, zinc, or lead. In general, growth varied with species and was dependent on the metal present and the concentration at which it was tested. Cadmium was found to exhibit the greatest toxicity followed by zinc and lead, respectively. In most cases, inhibition of growth was directly correlated with a decreased capacity to form traps. However, in a few cases, trap formation was inhibited either more or less than was growth. The activity of the collagenase was less sensitive than was growth or trap formation to heavy-metal inhibition. PMID- 16345646 TI - Effects of adaptation on biodegradation rates in sediment/water cores from estuarine and freshwater environments. AB - Experiments were devised to determine whether exposure to xenobiotics would cause microbial populations to degrade the compounds more rapidly during subsequent exposures. Studies were done with water/sediment systems (ecocores) taken from a salt marsh and a river. Systems were tested for adaptation to the model compounds methyl parathion and p-nitrophenol. CO(2) released from radioactive parent compounds was used as a measure of mineralization. River populations preexposed to p-nitrophenol at concentrations as low as 60 mug/liter degraded the nitrophenol much faster than did control populations. River populations preexposed to methyl parathion also adapted to degrade the pesticide more rapidly, but higher concentrations were required. Salt marsh populations did not adapt to degrade methyl parathion. p-Nitrophenol-degrading bacteria were isolated from river samples but not from salt marsh samples. Numbers of nitrophenol degrading bacteria increased 4 to 5 orders of magnitude during adaptation. Results indicate that the ability of populations to adapt depends on the presence of specific microorganisms. Biodegradation rates in laboratory systems can be affected by concentration and prior exposure; therefore, adaptation must be considered when such systems are used to predict the fate of xenobiotics in the environment. PMID- 16345647 TI - Mineralization of detrital lignocelluloses by salt marsh sediment microflora. AB - Specifically radiolabeled C-(cellulose)-lignocellulose and C-(lignin) lignocellulose were isolated from labeled cuttings of Spartina alterniflora (cordgrass) and Pinus elliottii (slash pine). These were used to estimate the rates of mineralization to CO(2) of lignocelluloses of estuarine and terrestrial origin in salt marsh estuarine sediments. The lignin moiety of pine lignocellulose was mineralized 10 to 14 times more slowly than that of Spartina lignocellulose, depending on the source of inoculum. Average values for percent mineralization after 835 h of incubation were 1.4 and 13.9%, respectively. For Spartina lignocellulose, mineralization of the cellulose moiety was three times faster than that of the lignin moiety. Average values for percent mineralization after 720 h of incubation were 32.1 and 10.6%, respectively. Lignocellulose and lignin contents of live pine and Spartina plants were analyzed and found to be 60.7 and 20.9%, respectively, for pine and 75.6 and 15.1%, respectively, for Spartina. PMID- 16345648 TI - Species composition and barotolerance of gut microflora of deep-sea benthic macrofauna collected at various depths in the atlantic ocean. AB - The bacterial flora of marine animals collected at depths of 570 to 2,446 m was examined for population size and generic composition, and the barotolerant characteristics of selected bacterial isolates were determined. Total numbers of culturable, aerobic, heterotrophic bacteria were found to be low in animals collected at the greatest ocean depths sampled in this study. Vibrio spp. were predominant in 10 of 15 samples examined, and Photobacterium spp. and yeasts were the major components of the remainder. Pseudomonas, Achromobacter, and Flavobacterium spp. comprised minor components of the gut flora of deep-sea fish. Forty-six pure cultures isolated from samples of seven animals were tested for growth or viability after incubation for 1 week under pressures ranging from 100 to 750 atm. Strains of bacteria isolated from samples of fish intestine were more barotolerant than those from the stomach (P<0.01). When incubated at a pressure of 600 atm, viability of bacterial cultures originally isolated from fish caught at a depth of 570 m was significantly decreased in comparison with viability of cultures from animals caught at depths of 1,393 and 2,446 m (P<0.01). From results of this study, it is concluded that the gut microflora of animals that dwell in the deeper regions of the ocean are adapted to an increased hydrostatic pressure environment, that is, the gut microflora is less inhibited by elevated hydrostatic pressure with increasing depth from which the host animal was collected. PMID- 16345649 TI - Measurement of mercury methylation in lake water and sediment samples. AB - Biological mercury methylation was assayed by a new radiochemical technique in the water column and sediments of a mercury-contaminated lake. In 24 weeks during 1979, there were three episodes of methylating activity in surface floc and in water, each lasting 3 to 5 weeks. Periods of methylation in the water column coincided with surface sediment methylation and appeared to be related to overall microbial activity. Mercury was actively methylated in the presence of bound sulfide. PMID- 16345650 TI - Appraisal of media and methods for assay of bacteriophages of lactic streptococci. AB - To assess the relative merits of tryptone yeast extract agar, the same medium unbuffered, and medium M17 for the assay of nine bacteriophages of lactic streptococci, comparative plaque counts were made with an overlay of 3 or 9 ml. Four of the phages exhibited no significant difference in plating efficiency between media. The effect of overlay volume varied from strain to strain and was different for different media. The 3-ml overlay created suboptimal atmospheric conditions for those strains which had a special requirement for CO(2). The use of a 9-ml overlay obviated the need to incubate plates under CO(2) and overcame the problems related to special calcium requirements when tryptone yeast extract agar was used. The organic buffer (disodium beta-glycerophosphate) was inhibitory to Streptococcus cremoris ML1 and showed no advantage over the inorganic phosphate buffer (K(2)HPO(4)) for most other strains. PMID- 16345651 TI - Attack on lignified grass cell walls by a facultatively anaerobic bacterium. AB - A filamentous, facultatively anaerobic microorganism that attacked lignified tissue in forage grasses was isolated from rumen fluid with a Bermuda grass containing anaerobic medium in roll tubes. The microbe, designated 7-1, demonstrated various colony and cellular morphologies under different growth conditions. Scanning electron microscopy revealed that 7-1 attacked lignified cell walls in aerobic and anaerobic culture. 7-1 predominately degraded tissues reacting positively for lignin with the chlorine-sulfite stain (i.e., sclerenchyma in leaf blades and parenchyma in stems) rather than the more resistant acid phloroglucinol-positive tissues (i.e., lignified vascular tissue and sclerenchyma ring in stems), although the latter tissues were occasionally attacked. Turbidimetric tests showed that 7-1 in anaerobic culture grew optimally at 39 degrees C at a pH of 7.4 to 8.0. Tests for growth on plant cell wall carbohydrates showed that 7-1 grew on xylan and pectin slowly in aerobic cultures but not with pectin and only slightly with xylan in anaerobic culture. 7-1 was noncellulolytic as shown by filter paper tests. The microbe used the phenolic acids sinapic, ferulic, and p-coumaric acids as substrates for growth; the more highly methoxylated acids were used more effectively. PMID- 16345652 TI - Effects of Pesticides on the Fatty Acid and Phospholipid Composition of Escherichia coli. AB - [This corrects the article on p. 232 in vol. 40.]. PMID- 16345653 TI - Denitrification in Low pH Spodosols and Peats Determined with the Acetylene Inhibition Method. AB - [This corrects the article on p. 235 in vol. 40.][This corrects the article on p. 236 in vol. 40.][This corrects the article on p. 238 in vol. 40.][This corrects the article on p. 239 in vol. 40.]. PMID- 16345654 TI - Antibiotic Resistances of Yogurt Starter Cultures Streptococcus thermophilus and Lactobacillus bulgaricus. AB - Twenty-nine strains of Lactobacillus bulgaricus and 15 strains of Streptococcus thermophilus were tested for resistance to 35 antimicrobial agents by using commercially available sensitivity disks. Approximately 35% of the isolates had uncharacteristic resistance patterns. PMID- 16345655 TI - Purification and Partial Characterization of an Aminopeptidase from Lactobacillus lactis. AB - A surface-bound aminopeptidase of Lactobacillus lactis cells was solubilized with lysozyme, and the extract was subjected to streptomycin sulfate precipitation, ammonium sulfate fractionation, chromatography on Sephadex G-100 and diethylaminoethyl-Sephadex A-50, and preparative polyacrylamide gel electrophoresis. The purified enzyme was homogeneous in disc electrophoretic analysis and consisted of a single polypeptide chain with a molecular weight of 78,000 to 81,000. The optimal pH and optimal temperature for enzyme activity were 6.2 to 7.2 and 47.5 degrees C, respectively, for l-lysine-4-nitroanilide as the substrate. The enzyme was activated by Co and Zn ions and inhibited by Cu, Hg, and Fe ions and by the metal-complexing reagents ethylenediaminetetraacetic acid, 1,10-phenanthroline, and alpha,alpha'-dipyridyl. Higher concentrations of substrate and hydrolysis products also inhibited the activity of the enzyme. The aminopeptidase had broad substrate specificity and hydrolyzed many amino acid arylamides and many peptides with unsubstituted NH(2)-terminal amino acids. PMID- 16345656 TI - Rocket Immunoelectrophoresis of the Entomocidal Parasporal Crystal of Bacillus thuringiensis subsp. kurstaki. AB - Rocket immunoelectrophoresis was used to quantitate the soluble parasporal crystal of Bacillus thuringiensis subsp. kurstaki. The method described is rapid, reliable, specific, and extremely accurate, and it can be used to measure crystal toxin in commercial microbial insecticides that contain a mixture of spores, vegetative cells, and carrier materials. PMID- 16345657 TI - Evidence for cometabolism in sewage. AB - A procedure was developed to demonstrate cometabolism in models of natural ecosystems. The procedure involves showing the formation of metabolic products in high yield and the lack of incorporation of substrate carbon into cellular constituents. Samples of four C-labeled herbicides (trifluralin, profluralin, fluchloralin, and nitrofen) were incubated with sewage aerobically and under discontinuous anaerobiosis for 88 days, and fresh sewage was added at intervals. Products were formed from each of the herbicides in nonsterile, but not in sterile, sewage. The yield of recovered products reached 87% for profluralin and more than 90% for fluchloralin and trifluralin, and the number of products ranged from 6 for nitrofen to 12 for fluchloralin. Concentrating the sewage microflora 40-fold greatly enhanced the rate of conversion. None of the radioactivity from the herbicide entered the nucleoside pool of the sewage microflora. The lack of incorporation of substrate carbon into cells and the almost stoichiometric conversion of the substrate to organic products indicate that members of the microbial community were cometabolizing the test compounds. PMID- 16345658 TI - Diversity and Dynamics of Indigenous Rhizobium japonicum Populations. AB - A simple method, based upon the separation of cellular proteins by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, has been devised for distinguishing between isolates of Rhizobium japonicum. Eleven laboratory strains, previously classified into five serogroups, were analyzed by gel electrophoresis. Groups determined subjectively according to protein patterns matched the serogroups, with one exception. Most strains within serogroups could be distinguished from one another. For studying the ecology of Rhizobium, an important advantage of this technique compared with serology or phage typing is that it discriminates among previously unencountered indigenous bacterial isolates as well as among known laboratory strains. SDS-gels were used to analyze the Rhizobium population of 500 nodules, sampled throughout the growing season, from soybeans at two different Wisconsin localities. Although the soybeans had been inoculated with laboratory strains of R. japonicum, indigenous R. japonicum predominated. At one location, 19 indigenous gel types were distinguished and classified mainly into four groups. At the other location, 18 gel types, falling mainly into three groups, were detected. The predominance of a particular group varied, in some cases dramatically, depending upon the time and depth of nodule formation. PMID- 16345659 TI - Bacterial degradation of dichloromethane. AB - Strain DM1, a facultative methylotrophic bacterium utilizing methanol, formate, mono-, di-, and trimethylamine, as well as dichloromethane as C1 substrates was isolated as an airborne contaminant. The organism is a strictly aerobic, gram negative, oxidase-positive short rod, motile by a single lateral flagellum. Enzyme assays in crude extracts suggested that it assimilates C1 compounds by the serine/isocitrate lyase-negative pathway. Experiments with extended cultures demonstrated that dichloromethane is a growth-inhibitory substrate. The maximum specific growth rate of 0.11 h was reached between 2 and 5 mM dichloromethane. The release of Cl from dichloromethane paralleled growth in extended and continuous cultures. Molar growth yields on methanol and on dichloromethane were 18.6 and 15.7 g/mol, respectively. Since attempts to demonstrate dehalogenation of dichloromethane by crude extracts failed, a dehalogenation assay with resting cells was developed. Maximum dehalogenating activity of cell suspensions was at pH 9.0. The reaction was partially and reversibly inhibited by anaerobiosis. During a shift of a chemostat culture from methanol to dichloromethane as the carbon source, the dehalogenating activity of resting cells was increased at least 500-fold. PMID- 16345660 TI - Action of douglas fir tussock moth larvae and their microflora on dietary terpenes. AB - A single type of bacterium, tentatively identified as a member of the genus Bacillus, was isolated from 2 of 20 midguts of Douglas fir tussock moth larvae being fed a diet of fir needles. No bacteria could be isolated from most midguts. Although spherically shaped bodies were present in the food bolus, these bodies, if microorganisms, could not be distinguished from spherical bodies associated with the plant tissue. The Douglas fir tussock moth dietary terpenes were altered during their passage through the insects, with two new terpenes being detected in the feces. One of these was identified as isoborneol. The relative significance of the insect and gut microflora with respect to terpene modification is unresolved. The well-established toxicity of terpenes may account for the near absence of common gut microflora in the insects. PMID- 16345661 TI - Prophage-Cured Derivatives of Streptococcus lactis and Streptococcus cremoris. AB - Prophage curing was achieved in Streptococcus lactis and Streptococcus cremoris, and the cured derivatives were shown to be indicators for their temperate bacteriophages. Relysogenization of these cured derivatives completed the first formal demonstration of the lysogenic state in lactic streptococci. PMID- 16345662 TI - Comparison of Two Alcohol Dehydrogenases in the Fungus Rhizopus javanicus. AB - Two alcohol dehydrogenases with different catalytic properties were separated from Rhizopus javanicus, and some of their immunological and molecular properties were compared. PMID- 16345663 TI - Fungal Pathogen of Cladophora glomerata (Chlorophyta). AB - A strain of Acremonium kiliense (Fungi Imperfecti) produced a water-soluble, dialyzable, heat-stable agent that rendered Cladophora glomerata (Chlorophyta) chlorotic and inhibited its growth. PMID- 16345664 TI - Interactions Between Light and Gas Vacuoles in Halobacterium salinarium Strain 5: Effect of Ultraviolet Light. AB - The potential light shielding by intracellular gas vacuoles in Halobacterium salinarium strain 5 was examined by looking at the ultraviolet light inactivation curves of both wild-type cells and mutants which are defective in the production of gas vacuoles. Whereas strains defective in gas vacuole production were slightly more sensitive to ultraviolet inactivation, no significant differences in ultraviolet sensitivity were seen, indicating that these subcellular inclusion bodies are not effective as light-shielding organelles. In addition, it was shown that ultraviolet light acts as a plasmid-curing agent in Halobacterium. PMID- 16345666 TI - Virulence Prediction of Yersinia enterocolitica by Pyrolysis Gas-Liquid Chromatography. AB - [This corrects the article on p. 648 in vol. 40.]. PMID- 16345665 TI - Rapid radiometric method for detection of salmonella in foods. AB - [This corrects the article on p. 223 in vol. 40.][This corrects the article on p. 225 in vol. 40.][This corrects the article on p. 226 in vol. 40.][This corrects the article on p. 229 in vol. 40.]. PMID- 16345667 TI - Analysis, Production, and Isolation of an Extracellular Laccase from Polyporus anceps. AB - Methods are described for the analysis, production, and isolation of laccase produced by a strain of Polyporus anceps. A simple quantitative colorimetric assay based on the oxidation of syringaldazine to syringaldazine quinone is described. Using a defined medium supplemented with the amino acids cysteine and histidine and with elevated phosphate, consistently high titers of laccase were obtained. The enzyme was isolated directly from fermentation medium by binding to diethylaminoethyl cellulose, and, once bound to the ion exchanger, it could be stored for 6 months at -70 degrees C with minimal loss of activity. The enzyme was quantitatively recovered from the resin by elution with 0.2 M phosphate buffer (pH 5.0). PMID- 16345668 TI - Distribution of iron in sphaerotilus and the associated inhibition. AB - The distribution of iron between the sheaths and the cells of iron-inhibited Sphaerotilus cultures was determined. The experiment was conducted with different soluble iron forms as inhibitors. The growth inhibition was found to be related to the iron sorbed by the cells rather than by the sheaths. At the 90% inhibition level, iron sorbed by the cells ranged from 13 to 15 mg/g of organism for all three inhibitors tested. For 50% inhibition, the iron sorbed by the cells ranged from 7 to 8 mg/g of organism. The iron sorbed by the sheaths varied widely, ranging from 23 to 118 mg/g of organism at the 90% inhibition level and from 11 to 61 mg/g at the 50% inhibition level. The degree of inhibition is closely related to the amount of iron sorbed by the cells, which in turn is a function of the type of iron compound or complex used. The solubility of the iron is a major consideration. PMID- 16345669 TI - Effects of heavy metal pollution on oak leaf microorganisms. AB - During the growing season, comparisons were made of the leaf surface microflora of (i) two groups of mature oak trees, one in the vicinity of a smelting complex contaminated by heavy metals and the other at a relatively uncontaminated site, and (ii) two groups of oak saplings at the uncontaminated site, one of which was sprayed with zinc, lead, and cadmium to simulate the heavy metal pollution from the smelter without the complicating effects of other pollutants. Total viable counts of bacteria, yeasts, and filamentous fungi (isolated by leaf washing) were generally little affected by the spraying treatment, whereas polluted leaves of mature trees supported fewer bacteria compared with leaves of mature trees at the uncontaminated site. Numbers of pigmented yeasts were lower on polluted oaks and on metal-dosed saplings compared with their respective controls. Polluted leaves of mature trees supported both greater numbers of Aureobasidium pullulans and Cladosporium spp. and a greater percentage of metal-tolerant fungi compared with oak leaves at the uncontaminated site. There were no significant overall differences in the degree of mycelial growth between the two groups of saplings or the mature trees. PMID- 16345670 TI - Production of nitrous oxide by ammonia-oxidizing chemoautotrophic microorganisms in soil. AB - Gas chromatographic studies showed that nitrous oxide was produced in each instance when sterilized (autoclaved) soil was incubated after treatment with ammonium sulfate and inoculation with pure cultures of ammonia-oxidizing chemoautotrophic microorganisms (strains of Nitrosomonas, Nitrosospira, and Nitrosolobus). Production of N(2)O in ammonium-treated sterilized soil inoculated with Nitrosomonas europaea increased with the concentration of ammonium and the moisture content of the soil and was completely inhibited by both nitrapyrin and acetylene. Similar effects of nitrapyrin, acetylene, ammonium concentration, and soil moisture content were observed in studies of factors affecting N(2)O production in nonsterile soil treated with ammonium sulfate. These observations support the conclusion that, at least under some conditions, most of the N(2)O evolved from soils treated with ammonium or ammonium-producing fertilizers is generated by chemoautotrophic nitrifying microorganisms during oxidation of ammonium to nitrite. PMID- 16345671 TI - Incorporation of P and Growth of Pseudomonad UP-2 on n-Tetracosane. AB - Cultures of the marine pseudomonad UP-2 growing on n-tetracosane contained both free cells and cells bound to the solid hydrocarbon. After separation by filtration through a Whatman no. 1 filter, the numbers of free and bound cells were estimated from the amount of P incorporated into each fraction and the determined value of P incorporation per viable cell in the filtrate (free cells). During the early exponential growth phase, over 80% of the cells were bound to large pieces of n-tetracosane; as the culture approached the stationary phase, the number of bound cells remained constant, whereas free cells continued to accumulate. Pulse-labeling experiments indicated that cells grew both on the surface of the solid and in the aqueous medium. During the growth cycle, a portion of the n-tetracosane which was initially nonfilterable was recovered in the filtrate in a form which was largely cell associated. This cell-associated n tetracosane was preferentially utilized and could completely account for the observed growth of free cells. PMID- 16345672 TI - Scanning Electron Microscopy of Rhizobium trifolii Infection Sites on Root Hairs of White Clover. AB - White clover root hairs which were inoculated with Rhizobium trifolii 4S (infectious strain) contained infection threads which were observed by light microscopy and scanning electron microscopy. Three morphological types of root hairs retaining infection threads were recognized. The bacteria were strongly attached between the surfaces of two plant cell walls as follows: between surfaces of a root hair tip curled back on itself, between a protuberance from a root hair and its cell surface, or between two root hair tips clinging together. An anatomical analysis documented the attachment site of the infection thread sheath from the inside of the root hair cell. PMID- 16345673 TI - Transformation assay for identification of psychrotrophic achromobacters. AB - The finding that many psychrotrophic, gram-negative, nonmotile, oxidase-positive coccobacilli (achromobacters) are competent for genetic transformation made possible the development of a transformation assay that permits recognition of genetically related strains. It has been demonstrated that 109 independently isolated achromobacters are genetically related since deoxyribonucleic acid samples from all of these organisms were able to transform a single competent auxotrophic strain to prototrophy. Genetically interacting bacteria included strains that lacked one or more of the characteristics typical for most achromobacters. An oxidase-negative mutant of one of these strains reacted positively in the transformation assay, unlike other oxidase-negative bacteria. Achromobacters were derived from fish, poultry, irradiated foods, seawater, and other sources. One strain previously classified as Micrococcus cryophilus has been shown to be related to the achromobacters. Two achromobacters had an optimum growth temperature of 35 degrees C and behaved as typical mesophiles. The moraxellae and Acinetobacter were shown to be unrelated to the achromobacters by using the transformation assay. The ready demonstration of genetic relatedness provides a new basis for taxonomic grouping of the psychrotrophic achromobacters. PMID- 16345674 TI - Progression of epiphytic microflora in wheat and alfalfa silages as observed by scanning electron microscopy. AB - Wheat and alfalfa silages were examined by scanning electron microscopy and standard methods of microbial enumeration. Epiphytic microflora were present at levels of 10 to 10/g in the fresh-cut plants. This flora was initially observed microscopically primarily on the surfaces. After 4 days of fermentation, lactic acid bacteria were observed on the surface in high concentrations near open stomata and throughout the interior mesophyll air sac spaces. At 4 days, populations on interior surfaces were restricted to the exterior surfaces of the air sacs. After 8 days the mesophyllic cells showed marked deterioration, and bacteria were observed on their inner surfaces. At 32 days, the end of the fermentation, vascular bundles and epidermal cells remained intact whereas stomata and mesophyllic cells were collapsed and often contained microorganisms. It is concluded that the interior of the leaves offers substantial nutritional and environmental advantages to epiphytic flora and is an important if not major deterioration site in fermented products. Since little deterioration of exterior surfaces was observed, these sites may play a minor role in supplying nutrients for microbial growth. PMID- 16345675 TI - Cellulose degradation by cellulose-clearing and non-cellulose-clearing brown-rot fungi. AB - Cellulose degradation by four cellulose-clearing brown-rot fungi in the Coniophoraceae-Coniophora prasinoides, C. puteana, Leucogyrophana arizonica, and L. olivascens-is compared with that of a non-cellulose-clearing brown-rot fungus, Poria placenta. The cellulose- and the non-cellulose-clearing brown-rot fungi apparently employ similar mechanisms to depolymerize cellulose; most likely a nonenzymatic mechanism is involved. PMID- 16345676 TI - Effects of pH and Sugar on Acetoin Production from Citrate by Leuconostoc lactis. AB - The relationship between acetoin production and citrate utilization in Leuconostoc lactis NCW1 was studied. In a complex medium the organism utilized citrate at neutral pH (initial pH, 6.3) and at acid pH (initial pH, 4.5) but produced nine times more acetoin at the latter pH. In resting cells the utilization of citrate was optimum at pH 5.3. Production of acetoin as a function of citrate utilization increased as the pH decreased, and at pH 4.3 all of the citrate utilized was recovered as acetoin. Glucose (10 mM) and lactose (10 mM) markedly stimulated citrate utilization but totally inhibited acetoin production in glucose- and lactose-grown cells. Addition of glucose to cells actively metabolizing citrate caused an immediate increase in citrate uptake and a reduction in the level of acetoin. The apparent K(m) values of lactic dehydrogenase for pyruvate were 1.05, 0.25, and 0.15 mM at pH 7.5, 6.5, and 5.0, respectively. Several heterofermentation intermediates inhibited alpha acetolactate synthetase and decarboxylase activities. The implications of these results in regulating acetoin formatin are discussed. PMID- 16345677 TI - Detection of Lipid Degradation Products in the Water of a Reservoir During a Bloom of Synura uvella. AB - A stripping method was used to determine the volatile compounds present in the raw water of a reservoir during a heavy bloom of Synura uvella. Besides numerous pollution products, a large number of previously unidentified algal excretion products were determined, including alkenes, alcohols, ketones, aldehydes, and nor-carotenoids. The approximate amounts of some important algal excretion products present in the raw water are stated. Their biosynthetic origin and possible function in the ecosystem are discussed. The offensive cod liver oil like odor which Synura imparts to the raw water was traceable to trans,cis-deca 2,4-dienal and minor amounts of trans,cis-hepta-2,4-dienal. PMID- 16345678 TI - Bacterial density in water determined by poisson or negative binomial distributions. AB - The question of how to characterize the bacterial density in a body of water when data are available as counts from a number of small-volume samples was examined for cases where either the Poisson or negative binomial probability distributions could be used to describe the bacteriological data. The suitability of the Poisson distribution when replicate analyses were performed under carefully controlled conditions and of the negative binomial distribution for samples collected from different locations and over time were illustrated by two examples. In cases where the negative binomial distribution was appropriate, a procedure was given for characterizing the variability by dividing the bacterial counts into homogeneous groups. The usefulness of this procedure was illustrated for the second example based on survey data for Lake Erie. A further illustration of the difference between results based on the Poisson and negative binomial distributions was given by calculating the probability of obtaining all samples sterile, assuming various bacterial densities and sample sizes. PMID- 16345679 TI - Surface-Active Lipids from Nocardia erythropolis Grown on Hydrocarbons. AB - Nocardia erythropolis (ATCC 4277) was grown in a 28-liter fermentor on mineral salts medium and 4% hydrocarbon. Extraction of the neutral lipids with pentane removed approximately 90% of the surface activity of the culture medium. The residual surface activity of the culture medium was attributed to the polar lipid fraction which was not extracted with pentane. Analysis of the pentane extracts with thin-layer chromatography showed the presence of four major compounds. A fatty alcohol reached a maximum concentration in the early log phase of growth and then decreased to the end of the fermentation. A monoglyceride, an ester, and a fatty acid appeared during the log phase of growth and continued to increase until the end of the fermentation. The fatty acids isolated from the culture grown on hexadecane had a carbon skeleton with the same length as the substrate, with 70% of the component as the saturated acid and 30% as a monounsaturated homolog. When isolated from a kerosene culture, the fatty acids consisted of a number of homologs from C(18) to C(20), including branched-chain and unsaturated acids, reflecting the distribution of the branched-chain isomers in the substrate. PMID- 16345680 TI - Formation of Short-Chain Fatty Acids from H(2) and CO(2) by a Mixed Culture of Bacteria. AB - The biological utilization of CO(2) and H(2) for the formation of short-chain fatty acids was studied by using a mixed culture of bacteria. Optimization of a medium was carried out in continuous culture to identify limiting factors which controlled growth and production of organic acids. The optimal pH for growth and acid production was 7.0 at 37 degrees C; the maximal cell concentration obtained was 5.9 g of cells per liter (dry weight), and the maximal amount of volatile acids formed was 4.7 g/liter, with acetic acid as the predominant acid. With the optimized medium, it was found that the rate of transfer of hydrogen or carbon dioxide, or both, from gas to liquid was the limiting factor which controlled growth and production of acids. PMID- 16345681 TI - Carrageenan from Eucheuma striatum (Schmitz) in Bacteriological Media. AB - The use of carrageenan from the red alga Eucheuma striatum as a possible substitute for bacteriological agar was investigated. The carrageenan medium was compared with several commercially available media in terms of both their physical properties and the colony characteristics of 16 microorganisms grown on the media. PMID- 16345682 TI - Influence of Nutritional Conditions on Production of l-Glutamine by Flavobacterium rigense. AB - The nutritional conditions for the production of l-glutamine by Flavobacterium rigense strain 703 were investigated. The optimum concentration of ammonia for achieving the highest yield of l-glutamine (25 mg/ml of broth) was relatively broad, from 0.9 to 1.6%, whereas fumaric acid had a narrow optimum range, near 5.5%. High concentration of inorganic ions such as chloride or sulfate ion clearly inhibited cell growth. Therefore, ammonium salts other than (NH(4))(2) fumarate were unsuitable for the highest production. The optimum concentration of (NH(4))(2)-fumarate was 7%. To reduce the concentration of fumaric acid in the medium, many substances were evaluated as substitutes. The fumaric acid concentration required for highest l-glutamine yield could not be replaced by any one of the compounds tested. However, part of fumaric acid could be replaced with succinic acid and cupric ion; 4% (NH(4))(2)-fumarate plus 2.5% succinic acid or 5% (NH(4))(2)-fumarate plus 1 mM cupric ion produced results similar to 7% (NH(4))(2)-fumarate in the fermentation medium. PMID- 16345683 TI - Yeasts from marine and estuarine waters with different levels of pollution in the state of rio de janeiro, Brazil. AB - Yeast counts were made at 24 marine and estuarine sites in the vicinity of Rio de Janeiro, Brazil. Mean salinities of estuarine sites ranged from 14.2 to 27.4 per thousand, and mean temperatures ranged from 25 to 28 degrees C. Total coliform counts varied from 80% above 100,000 colony-forming units (CFU)/100 ml at heavily polluted sites to 100% below 100 CFU/100 ml at unpolluted sites. Total yeast counts above 100 CFU/100 ml were typical of heavily and moderately polluted water but atypical of lightly polluted and unpolluted water. Mean total yeast counts were 2,880 CFU/100 ml for heavily polluted sites, 202 CFU/100 ml for moderately polluted sites, and 3 CFU/100 ml for lightly polluted and unpolluted sites. Total yeast counts had a positive response to increased pollution levels, and Candida krusei and phenotypically similar yeasts as a group were prevalent in polluted estuarine water but rare in unpolluted seawater. The 549 strains of yeasts and yeast-like organisms isolated were grouped into 67 species, of which the 21 most prevalent made up 86% of the total yeast population. The prevalent genera in the polluted estuary were Candida, Rhodotorula, Torulopsis, Hanseniaspora, Debaryomyces, and Trichosporon. PMID- 16345684 TI - Comparing time course profiles of immediate acetylene reduction by grasses and legumes. AB - The time course profiles of C(2)H(2) reduction by intact Scirpus olneyi (bulrush), Oryza sativa (rice) and Spartina alterniflora (cordgrass) with roots in atmospheres of N(2) and 30-day-old Glycine max (soybean) in air were all immediately linear. This is the first report of immediately linear rates of C(2)H(2) reduction by grass roots removed from soil. The immediately linear profile of C(2)H(2) reduction by soil-free grass roots was achieved by preventing contact between the roots and air. Roots of soybeans and S. olneyi receiving pretreatments of O(2) above normal environmental levels for 15 min before assay exhibited a short delay in C(2)H(2) reduction. These initially nonlinear rates of C(2)H(2) reduction are attributable to transient O(2) inhibition of nitrogenase. Initial nonlinear rates of C(2)H(2) reduction were also observed with immature soybean plants and with intact plant assays of O. sativa and S. olneyi in which C(2)H(2) was injected into cylinders surrounding the plant tops. These results indicate that, apart from O(2) inhibition of nitrogenase, the diffusion of C(2)H(2) and C(2)H(4) between the nitrogen-fixing sites and the sampling ports may cause initial nonlinear rates of C(2)H(2) reduction. We conclude that in situ plant-associated nitrogenase activity should result in immediate reduction of C(2)H(2) and that linear rates are observed when the proper assay conditions are used. Our data suggest that nitrogen fixation is closely associated with the roots of S. olneyi, O. sativa, and S. alterniflora growing in salt marsh sediment. PMID- 16345685 TI - Incidence of Plasmids in Marine Vibrio spp. Isolated from an Oil Field in the Northwestern Gulf of Mexico. AB - Presumptive marine Vibrio spp. were collected from an operational oil field and control site located in the northwestern Gulf of Mexico. Of 440 isolates analyzed for the presence of extrachromosomal deoxyribonucleic acid elements or plasmids by using the cleared lysate and agarose gel techniques, 31% showed distinct plasmid bands on agarose gels. A majority of the plasmids detected were estimated to have molecular masses of 10 x 10 or less. Multiple plasmids were observed in approximately half of the plasmid-containing strains. A number of isolates contained plasmids with similar banding and mobility patterns. The oil field area had noticeably more plasmid-containing strains (35 versus 23% in the control site) and a greater number of plasmids per plasmid-containing strain (an average of 2.5 plasmids, versus 1.5 in the control site). Oil field discharges might have resulted in increased plasmid incidence and diversity. PMID- 16345686 TI - Rates of microbial transformation of polycyclic aromatic hydrocarbons in water and sediments in the vicinity of a coal-coking wastewater discharge. AB - To facilitate predictions of the transport and fate of contaminants at future coal conversion facilities, rates of microbial transformation of polycyclic aromatic hydrocarbons were measured in stream water and sediment samples collected in the vicinity of a coal-coking treated wastewater discharge from November 1977 through August 1979. Six radiolabeled polycyclic aromatic hydrocarbons were incubated with sediment and water samples; CO(2), cell-bound C, and polar transformation products were isolated and quantified. Whereas CO(2) and bound C were major transformation products in sediment assays, soluble polar C dominated transformation in water samples. Mean rate constants (measured at 20 degrees C) in sediments collected downstream from the effluent outfall were 7.8 x 10 h (naphthalene), 1.6 x 10 h (anthracene), and 3.3 x 10 h [benz(a)anthracene], which corresponded to turnover times of 13, 62, and 300 h, respectively. No unequivocal evidence for transformation of benzo(a)pyrene or dibenz(a,h)anthracene was obtained. Only naphthalene and anthracene transformations were observed in water samples; rate constants were consistently 5- and 20-fold lower, respectively, than in the corresponding sediment samples. The measured rate constants for anthracene transformation in July 1978 sediment samples were not related to total heterotroph numbers. In late July 1978, the effluent was diverted from the primary study area; however, no differences were observed either in transformation rate constants or in the downstream/upstream sediment rate constant ratio. These results are consistent with the hypothesis that continuous inputs of polycyclic aromatic hydrocarbons result in an increased ability within a microbial community to utilize certain polycyclic aromatic hydrocarbons. However, because transformation rates remained elevated for more than 1 year after removal of the polycyclic aromatic hydrocarbon source, microbial communities may shift only slowly in response to changes in polycyclic aromatic hydrocarbon concentrations. PMID- 16345687 TI - Root-associated n(2) fixation (acetylene reduction) by enterobacteriaceae and azospirillum strains in cold-climate spodosols. AB - N(2) fixation by bacteria in associative symbiosis with washed roots of 13 Poaceae and 8 other noncultivated plant species in Finland was demonstrated by the acetylene reduction method. The roots most active in C(2)H(2) reduction were those of Agrostis stolonifera, Calamagrostis lanceolata, Elytrigia repens, and Phalaris arundinacea, which produced 538 to 1,510 nmol of C(2)H(4).g (dry weight). h when incubated at pO(2) 0.04 with sucrose (pH 6.5), and 70 to 269 nmol of C(2)H(4). g (dry weight).h without an added energy source and unbuffered. Azospirillum lipferum, Enterobacter agglomerans, Klebsiella pneumoniae, and a Pseudomonas sp. were the acetylene-reducing organisms isolated. The results demonstrate the presence of N(2)-fixing organisms in associative symbiosis with plant roots found in a northern climatic region in acidic soils ranging down to pH 4.0. PMID- 16345688 TI - Utilization of low concentrations of starch by a flavobacterium species isolated from tap water. AB - Experiments in well-cleaned glass flasks revealed that addition of starch in concentrations of 10 and 25 mug of substrate C per liter to the filtrate of slow sand filters stimulated the development of a yellow-pigmented bacterium which was identified as a Flavobacterium species. The isolate was able to multiply in tap water without substrates added, but addition of starch and glucose in amounts as low as 1 mug of substrate C per liter clearly enhanced growth. The substrate affinities of the Flavobacterium for these compounds were 3.9 mug of starch C and 3.3 mug of glucose C per liter. The results of this study indicate that microorganisms which rapidly utilize starch at a level of a few micrograms per liter commonly occur in water. PMID- 16345689 TI - High Production of beta-Glucosidase in Schizophyllum commune: Isolation of the Enzyme and Effect of the Culture Filtrate on Cellulose Hydrolysis. AB - Optimization experiments with response surface statistical analysis were performed with Schizophyllum commune to obtain high beta-glucosidase yields. The factors in the optimization experiment were the concentrations of cellulose, peptone, and KH(2)PO(4). Their optimal values were 3.2, 3.0, and 0.2 g/100 ml, respectively. Enzyme assays revealed very high beta-glucosidase (22.2 U/ml) and cellobiase (68.9 U/ml) yields. The avicelase yield was low as compared with that from Trichoderma reesei. Mixtures of S. commune and T. reesei culture filtrates caused faster and more extensive saccharification of Avicel than could be achieved by either filtrate alone. A beta-glucosidase was isolated and purified from the optimized culture filtrate of S. commune. The electrophoretic mobility of the purified beta-glucosidase indicated a molecular weight of 97,000. The amino acid composition was similar to that of beta-glucosidase from T. reesei. The acidic (aspartate and glutamate) residues or their amides or both made up approximately 20% of the protein. The NH(2)-terminal amino acid of the enzyme was histidine. PMID- 16345690 TI - Effect of protozoa on bacterial degradation of an aromatic compound. AB - Prototrophic and growth factor-requiring strains of Alcaligenes spp. were used to study the effect of a protozoan, Tetrahymena pyriformis, on the degradation of p aminobenzoate. The protozoan inhibited activity of the prototrophic bacterium by reducing its population size. For the growth factor-requiring strain of Alcaligenes, T. pyriformis provided the required growth factors so that the predator permitted the bacteria to grow and to continue p-aminobenzoate degradation. T. pyriformis inhibited bacterial activity when the amino acid supply was in excess, but activity of the auxotrophic strain of Alcaligenes was stimulated by the protozoan when the amino acid supply was limiting, although the bacterial population size was reduced by the protozoan. PMID- 16345692 TI - Microbiota Associated with the Gastrointestinal Tract of the Common House Cricket, Acheta domestica. AB - The location and morphology of the bacteria associated with the gastrointestinal tract of Acheta domestica were studied, and these bacteria were partially characterized. Bacteria were associated with the peritrophic membrane in the midgut and with the gut wall and cuticular structures of the hindgut. No bacteria were associated with the fat bodies. Colony-forming unit determinations indicated that there were three times more cultivatable bacteria in the hindgut than in the midgut. Of these bacteria, 40 to 85% cleared uric acid anaerobically, and 90 to 100% cleared uric acid aerobically. Of the 25 isolates obtained, 21 belonged to the genera Citrobacter, Klebsiella, Yersinia, Bacteroides, and Fusobacterium. PMID- 16345691 TI - Microbial Cells as Biosorbents for Heavy Metals: Accumulation of Uranium by Saccharomyces cerevisiae and Pseudomonas aeruginosa. AB - Uranium accumulated extracellularly on the surfaces of Saccharomyces cerevisiae cells. The rate and extent of accumulation were subject to environmental parameters, such as pH, temperature, and interference by certain anions and cations. Uranium accumulation by Pseudomonas aeruginosa occurred intracellularly and was extremely rapid (<10 s), and no response to environmental parameters could be detected. Metabolism was not required for metal uptake by either organism. Cell-bound uranium reached a concentration of 10 to 15% of the dry cell weight, but only 32% of the S. cerevisiae cells and 44% of the P. aeruginosa cells within a given population possessed visible uranium deposits when examined by electron microscopy. Rates of uranium uptake by S. cerevisiae were increased by chemical pretreatment of the cells. Uranium could be removed chemically from S. cerevisiae cells, and the cells could then be reused as a biosorbent. PMID- 16345693 TI - Symbiotic utilization of polyvinyl alcohol by mixed cultures. AB - Polyvinyl alcohol (PVA)-utilizing cultures were obtained from various sources. They were mixed cultures even after cyclical transfer to liquid and plate media with PVA as a sole source of carbon. Component bacteria were isolated from the several mixed cultures, and it was shown that PVA was utilized symbiotically by two bacterial members which could not utilize PVA in each respective pure culture. From a mixed culture, strains VM15, VM15A (Pseudomonas putida) and VM15C (Pseudomonas sp.) were isolated as members essential for PVA utilization. VM15C was the predominant strain in the mixed-culture population and produced PVA degrading enzyme. The culture supernatant of VM15A enabled VM15C to grow on PVA. VM15A was presumed to supply VM15C with a unique growth stimulant which was distinct from usual growth factors. PMID- 16345694 TI - Disruption of the primary fouling sequence on fiber glass-reinforced plastic submerged in the marine environment. AB - Fiber glass-reinforced plastic immersed in an experimental estuarine mesocosm fouled at estimated rates of 0.5, 5.5, and 18.8 ng (wet weight) mm day over days 0 to 2, 2 to 6, and 6 to 14, respectively. Protists, dominated by diatoms, which developed between days 3 and 6 and covered 90% of the undisturbed surface in 2 weeks, were effectively removed by twice-weekly brushing of the surface to maintain an immature 3-day bacterial film which covered 12% or less of the surface and had a biomass 3 orders of magnitude smaller than surfaces with 2 weeks' unrestricted fouling. Direct brushing of the fiber glass-reinforced plastic tank walls of experimental estuarine mesocosms minimized the "wall effect" by keeping a surface that maintained a low biomass of a slowly accumulating bacterial film rather than a surface which supported the more rapid accumulation of protists which in turn may induce the settlement of invertebrates and macrophytes. PMID- 16345695 TI - Iron bacteria in drinking-water distribution systems: elemental analysis of gallionella stalks, using x-ray energy-dispersive microanalysis. AB - "Iron" bacteria belonging to the genus Gallionella were observed by scanning electron microscopy in water samples and attached to pipe surfaces in a Southern California drinking-water distribution system. The cells were recognized by their characteristic elongated helical stalks composed of numerous intertwined microfibrils. Many of the stalks were partially coated with insoluble ferric salt deposits. Stalks recovered directly from water samples were analyzed for their elemental composition by using X-ray energy-dispersive microanalysis. Silicon, aluminum, calcium, and iron were the predominant elements present in the stalks. Smaller quantities of the elements phosphorous, sulfur, chlorine, copper, and zinc were also detected. Manganese, though present in measurable quantities in the water supply, was not detected in the stalks, suggesting that this organism is unable to utilize this element as an electron donor. This represents the first such analysis of Gallionella stalks recovered from environmental samples without prior subculturing in artificial laboratory media. PMID- 16345696 TI - Effect of dietary monensin or chlortetracycline on methane production from cattle waste. AB - Wastes from feedlot cattle fed finishing diets containing either monensin, chlortetracycline, or no antibiotic were investigated as substrates for methane production. We used continuously mixed anaerobic fermentors with 3-liter working volumes at 35 and 55 degrees C; these fermentors were fed once per day. Within a few days after waste from animals fed monensin was added, the volume of methane produced began to decrease in the 55 degrees C fermentors. After 9 days of daily feeding, methane production was severely inhibited, the pH dropped from 7.6 to 5.9, and the concentration of volatile acids increased from 543 to 6,300 mg/liter (as acetate). Although additions of waste from cattle fed monensin were discontinued after 9 days, the fermentors did not resume gas production within 8 weeks. The addition of waste from cattle which had been fed chlortetracycline reduced the methane production rate approximately 20%; however, pH and volatile acid values were comparable to control fermentor values after 40 days. Similar effects were observed with the 35 degrees C fermentors. In a batch fermentation experiment in which 50-g portions of volatile solids from waste of animals fed monensin, chlortetracycline, or no antibiotics were added to fermentors, monensin delayed the onset of methane production for about 40 days, but then these fermentors began to produce methane at a rate comparable to the control rate. The ultimate methane yields from the three types of waste after 180 days were not significantly different. These studies indicate that monensin has a detrimental effect on the conversion of feedlot wastes to methane, unless microorganisms can be adapted to the levels that are present in these wastes. PMID- 16345697 TI - Formation of bacterial microcolonies on feed particles in the rumen. AB - Examination of particulate feed that had been digested in vivo in the rumen, and of the leaves of specific legumes that had been digested in vitro by a mixed population of rumen bacteria, showed that very extensive glycocalyx-enclosed bacterial microcolonies developed on many of the available surfaces. Some of these adherent bacteria colonized a surface almost exclusively and attracted another specific type of bacteria as the second members of a distinct morphological consortium. The true extent of the exopolysaccharide glycocalyces of these adherent rumen bacteria was seen in cases where the fibers were attached at multiple points, and their role in microcolony formation and adhesion could be unequivocally ascribed. PMID- 16345698 TI - In vitro nitrogen fixation by two actinomycete strains isolated from casuarina nodules. AB - Acetylene reduction activity was demonstrated in pure cultures of two actinomycete strains isolated from nodules of Casuarina equisetifolia. This activity was comparable to that of free-living Rhizobium strains, but appeared to be less sensitive to pO(2) and more sensitive to the presence of combined nitrogen. PMID- 16345699 TI - Antibiotic sensitivities in vitro of diverse spiroplasma strains associated with plants and insects. AB - Fifteen spiroplasma strains, representing five serological subgroups classified in three distinct serogroups, and four strains unassigned to serogroups were examined for sensitivity to antibiotics. The data contribute to the characterization of spiroplasmas and enlarge comparisons between plant pathogenic strains and strains that are evidently a part of the normal epiphytic microflora. PMID- 16345700 TI - Development of a Defined Medium for Growth of Cercospora rosicola Passerini. AB - A simple synthetic liquid medium containing a single amino acid, glucose, salts, trace metals, and thiamine was developed for cultivation of Cercospora rosicola Passerini. Thiamine was shown to be important to growth. Culture of C. rosicola Passerini in a chemically defined medium makes possible studies of (+)-abscisic acid biosynthesis and regulation. PMID- 16345701 TI - Inorganic carbon limitation and chemical composition of two freshwater green microalgae. AB - Two freshwater chlorophytes, Chlorella vulgaris and Scenedesmus obliquus, were grown in inorganic carbon-limited continuous cultures in which HCO(3) was the sole source of inorganic carbon. The response of the steady-state growth rate to the external total inorganic carbon concentration was reasonably well described by the Monod equation; however, the response to the internal nutrient concentration was only moderately well represented by the Droop equation when the internal carbon concentration was defined on a cellular basis. The Droop equation was totally inapplicable when total biomass (dry weight) was used to define internal carbon because the ratio of carbon to dry weight did not vary over the entire growth rate spectrum. In batch cultures, maximum growth rates were achieved at the CO(2) levels present in atmospheric air and at HCO(3) concentrations of 3 mM. No growth was observed at 100% CO(2). Both nitrogen uptake and chlorophyll synthesis were tightly coupled to carbon assimilation, as indicated by the constant C/N and C/chlorophyll ratios found at all growth rates. The main influence of inorganic carbon limitation appears to be not on the chemical structure of the biomass, but rather on cell size; higher steady-state growth rates lead to bigger cells. PMID- 16345702 TI - Production of Skatole and para-Cresol by a Rumen Lactobacillus sp. AB - The objective of this study was to examine the substrate specificity of several ruminal strains of a Lactobacillus sp. which previously was shown to produce skatole (3-methylindole) by the decarboxylation of indoleacetic acid. A total of 13 compounds were tested for decarboxylase activity. The Lactobacillus strains produced p-cresol (4-methylphenol) by the decarboxylation of p hydroxyphenylacetic acid, but did not produce either o-cresol or m-cresol from the corresponding hydroxyphenylacetic acid isomers. These strains also decarboxylated 5-hydroxyindoleacetic acid to 5-hydroxyskatole and 3,4 dihydroxyphenylacetic acid to methylcatechol. Skatole and p-cresol were produced in a 0.5:1 ratio, when indoleacetic acid and p-hydroxyphenylacetic acid were combined in equimolar concentrations. Competition studies with indoleacetic acid and p-hydroxyphenylacetic acid suggested that two different decarboxylating enzymes are involved in the production of skatole and p-cresol by these strains. This is the first demonstration of both skatole production and p-cresol production by a single bacterium. PMID- 16345703 TI - Deoxyribonucleic Acid homology among lactic streptococci. AB - A comparison was made by deoxyribonucleic acid homology of 45 strains of lactic streptococci, using two strains of Streptococcus cremoris and three strains of Streptococcus lactis as reference strains. All S. cremoris strains were grouped together by deoxyribonucleic acid homology. S. lactis strains formed a second group, except that three strains of S. lactis showed a high degree of homology with S. cremoris strains. The three Streptococcus diacetylactis strains could not be differentiated from S. lactis strains. In spite of these differences between S. lactis and S. cremoris strains, the majority of S. cremoris, S. lactis, and S. diacetylactis strains studied had at least 50% of their base sequences in common. In contrast, Streptococcus thermophilus strains generally showed little relationship with the other strains of lactic streptococci. The relevance of these findings to the selection of starter strains for cheese making is discussed. PMID- 16345704 TI - Purification and Some Properties of Diplococcin from Streptococcus cremoris 346. AB - Eleven of 150 Streptococcus cremoris strains examined produced the bacteriocin diplococcin. The diplococcin activity spectrum was restricted to S. cremoris and Streptococcus lactis strains, and none of a wide range of other gram-positive or gram-negative strains were inhibited. The diplococcin produced by S. cremoris 346 was purified by ammonium sulfate precipitation and column chromatography. Purified diplococcin was very unstable at room temperature and lost 75% of its activity after heating at 100 degrees C for 1 min. The proteolytic enzymes trypsin, pronase, and alpha-chymotrypsin completely inactivated diplococcin. The amino acid composition showed a high content of acidic and neutral acids and a correspondingly low content of basic amino acids, including one residue of ornithine per mole. From the amino acid analysis a molecular weight of 5,300 was estimated. Diplococcin was readily distinguished from the S. lactis bacteriocin nisin by its restricted activity spectrum, its biological properties, and by cross-reaction experiments. PMID- 16345705 TI - Nodulation of acacia species by fast- and slow-growing tropical strains of Rhizobium. AB - Thirteen Acacia species were classified into three groups according to effective nodulation response patterns with fast- and slow-growing tropical strains of Rhizobium. The first group nodulated effectively with slow-growing, cowpea-type Rhizobium strains; the second, with fast-growing Rhizobium strains; and the third, with both fast- and slow-growing Rhizobium strains. The Rhizobium requirements of the Acacia species of the second group were similar to those of Leucaena leucocephala. PMID- 16345706 TI - Poplar lignin decomposition by gram-negative aerobic bacteria. AB - Eleven gram-negative aerobic bacteria (Pseudomonadaceae and Neisseriaceae) out of 122 soil isolates were selected for their ability to assimilate poplar dioxane lignin without a cosubstrate. Dioxane lignin and milled wood lignin degradation rates ranged between 20 and 40% of initial content after 7 days in mineral medium, as determined by a loss of absorbance at 280 nm; 10 strains could degrade in situ lignin, as evidenced by the decrease of the acetyl bromide lignin content of microtome wood sections. No degradation of wood polysaccharides was detected. Lignin biodegradation by Pseudomonas 106 was confirmed by CO(2) release from labeled poplar wood, although in lower yields compared with results obtained through chemical analysis based on acetyl bromide residual lignin determination. PMID- 16345707 TI - Plant yield and nitrogen content of a digitgrass in response to azospirillum inoculation. AB - Two Australian soils, a vertisol (pH 6.8, 0.299% N) and a sandy yellow podzol (pH 6.2, 0.042% N), were used with digitgrass, Digitaria sp. X46-2 (PI 421785), in a growth room experiment. Comparisons were made between plants inoculated with live and autoclaved bacterial suspensions of Australian and Brazilian isolates of Azospirillum brasilense. Seedlings were inoculated on days 10 and 35. Acetylene reducing activity was measured five times during the experiment. Dry matter yields of the digitgrass on the podzol (low N) inoculated with live bacteria were 23% higher than those of the controls. On the vertisol (high N), yield increases from inoculation with live bacteria were 8.5%. The higher-yielding plants had significantly lower percent nitrogen, but when total nitrogen of the tops was calculated, the inoculated plants had a higher total N than did the controls (P=0.04). Acetylene-reducing activity was variable in the experiment, ranging from 0.5 to 11.9 mumol of C(2)H(4) core day. Live bacterial treatment induced a proliferation of roots, possible earlier maturity, higher percent dry matter, and a higher total N in the tops. PMID- 16345708 TI - Anaerobic Degradation of Lactate by Syntrophic Associations of Methanosarcina barkeri and Desulfovibrio Species and Effect of H(2) on Acetate Degradation. AB - When grown in the absence of added sulfate, cocultures of Desulfovibrio desulfuricans or Desulfovibrio vulgaris with Methanobrevibacter smithii (Methanobacterium ruminantium), which uses H(2) and CO(2) for methanogenesis, degraded lactate, with the production of acetate and CH(4). When D. desulfuricans or D. vulgaris was grown in the absence of added sulfate in coculture with Methanosarcina barkeri (type strain), which uses both H(2)-CO(2) and acetate for methanogenesis, lactate was stoichiometrically degraded to CH(4) and presumably to CO(2). During the first 12 days of incubation of the D. desulfuricans-M. barkeri coculture, lactate was completely degraded, with almost stoichiometric production of acetate and CH(4). Later, acetate was degraded to CH(4) and presumably to CO(2). In experiments in which 20 mM acetate and 0 to 20 mM lactate were added to D. desulfuricans-M. barkeri cocultures, no detectable degradation of acetate occurred until the lactate was catabolized. The ultimate rate of acetate utilization for methanogenesis was greater for those cocultures receiving the highest levels of lactate. A small amount of H(2) was detected in cocultures which contained D. desulfuricans and M. barkeri until after all lactate was degraded. The addition of H(2), but not of lactate, to the growth medium inhibited acetate degradation by pure cultures of M. barkeri. Pure cultures of M. barkeri produced CH(4) from acetate at a rate equivalent to that observed for cocultures containing M. barkeri. Inocula of M. barkeri grown with H(2)-CO(2) as the methanogenic substrate produced CH(4) from acetate at a rate equivalent to that observed for acetate-grown inocula when grown in a rumen fluid-vitamin-based medium but not when grown in a yeast extract-based medium. The results suggest that H(2) produced by the Desulfovibrio species during growth with lactate inhibited acetate degradation by M. barkeri. PMID- 16345709 TI - Growth Inhibition of Streptomyces Species by l-Serine and Its Effect on Tetracycline Biosynthesis. AB - The addition of serine to minimal medium inhibited the growth of Streptomyces aureofaciens and Streptomyces rimosus. Both the outgrowth of spores and the growth of vegetative cells were inhibited by l-serine. This effect was independent of the carbon source used. In rich nutrient medium, however, the serine effect was not observed. The presence of glycine and methionine in minimal medium reversed the growth inhibition imposed by serine, suggesting that a metabolic block related to the synthesis of these two amino acids was involved. A serine-tolerant mutant of S. aureofaciens isolated after ultraviolet irradiation showed a level of serine deaminase comparable to that of the wild-type strain, which indicated that tolerance to serine was not due to the presence of a more active deaminating enzyme in the mutant. Serine markedly reduced tetracycline and oxytetracycline biosynthesis with the parental strains of Streptomyces spp. The serine-tolerant mutant, however, produced almost the same amount of tetracycline in the presence or absence of serine. The final cell population in fermentation broth was not significantly reduced by l-serine, and the addition of glycine and methionine did not increase the tetracycline yields, which suggested that l serine inhibition of antibiotic biosynthesis was by a mechanism different from that related to growth inhibition. PMID- 16345710 TI - Purification and Characterization of an Autolysin from Clostridium acetobutylicum. AB - A proteinaceous substance with antibiotic-like activity, resembling that of a bacteriocin, was isolated from an industrial-scale acetone-butanol fermentation of Clostridium acetobutylicum. The substance, purified by acetone precipitation, diethylaminoethyl cellulose chromatography, and polyacrylamide gel electrophoresis, was characterized as a glycoprotein with a molecular weight of 28,000. The glycoprotein was partially inactivated by certain protease enzymes. It had no effect on deoxyribonucleic acid, ribonucleic acid, or protein synthesis, and it did not result in the loss of intracellular adenosine triphosphate. The glycoprotein lysed sodium dodecyl sulfate-treated cells and cell wall preparations, and therefore it is referred to as an autolysin. The autolysin gene appeared to be chromosomal since plasmid deoxyribonucleic acid was not detected in the C. acetobutylicum strain. PMID- 16345711 TI - Temporal changes in a pink feedlot lagoon. AB - Temporal changes in a feedlot drainage lagoon with a predominance of the phototrophic purple sulfur bacterium, Thiopedia rosea, were investigated during a 3-year period. The surface protein and bacteriochlorophyll concentrations, which indirectly measure T. rosea abundance, peaked annually during the fall months and coincided with the intensity of pink coloration. Surface bacteriochlorophyll concentration correlated with pH, alkalinity, and protein. The pH range was optimal for the survival of T. rosea. Surface sulfide concentration, which increased over the winter and early spring, reached low levels during the fall months. The most striking pattern to emerge was the marked increase in sulfate concentration that occurred each fall and winter. The protein peaks, which preceded the sulfate peaks, were indicative of the sulfate concentrations that would follow. During 1977 and 1978, the lagoon was essentially anaerobic and provided adequate growth conditions for T. rosea. Above-average precipitation during early 1979 raised the water level and altered the chemistry of the lagoon. Dissolved oxygen was higher during the final year, and, concurrently, concentrations of bacteriochlorophyll declined. Aeration of the lagoon resulted in a decrease in T. rosea. PMID- 16345712 TI - Selective-differential medium for isolation and differentiation of pectinatus from other brewery microorganisms. AB - An agar medium, LL-agar (lactate-lead acetate) was designed to selectively differentiate members of the genus Pectinatus (S. Y. Lee, M. S. Mabee, and N. O. Jangaard, Int. J. Syst. Bacteriol. 28:582-594, 1978; S. Y. Lee, M. S. Mabee, N. O. Jangaard, and E. K. Horiuchi, J. Inst. Brew. 86:28-30, 1980) from other brewery microorganisms. Selectivity was achieved by the use of sodium lactate as the sole source of carbon and phenylethyl alcohol as an inhibitor for aerobic gram-negative bacteria and yeast. Differentiation was established by the introduction of lead acetate into the medium, which reacted with the H(2)S liberated by Pectinatus and resulted in a blackening of the Pectinatus colonies while the other brewery organisms, when present, remained white. In combination with the Lee tube (J. E. Ogg, S. Y. Lee, and B. J. Ogg, Can. J. Microbiol. 25:987 990, 1979) and this medium, isolation of Pectinatus organisms from beer samples was accomplished with convenience and simplicity. PMID- 16345713 TI - Preparation of uridine diphosphate-N-acetylgalactosamine from uridine diphosphate N-acetylglucosamine by using microbial enzymes. AB - A method was developed for the large scale preparation of uridine diphosphate-N acetylgalactosamine (UDP-GalNAc) from uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) by means of microbial enzymes. With Bacillus subtilis cell-free extract as a source of UDP-GlcNAc 4-epimerase, about 35% of the UDP-GlcNAc added was converted to UDP-GalNAc. After the residual UDP-GlcNAc was degraded to uridine triphosphate and N-acetylglucosamine-1-phosphate with a protamine-treated extract of bakers' yeast as a source of UDP-GlcNAc pyrophosphorylase, UDP-GalNAc was separated by anion-exchange column chromatography. The nucleotide was recovered by adsorption on charcoal and elution with ammoniacal ethanol. The final yield was about 100 mumol. PMID- 16345714 TI - Anaerobic oxidation of acetylene by estuarine sediments and enrichment cultures. AB - Acetylene disappeared from the gas phase of anaerobically incubated estuarine sediment slurries, and loss was accompanied by increased levels of carbon dioxide. Acetylene loss was inhibited by chloramphenicol, air, and autoclaving. Addition of C(2)H(2) to slurries resulted in the formation of CO(2) and the transient appearance of C-soluble intermediates, of which acetate was a major component. Acetylene oxidation stimulated sulfate reduction; however, sulfate reduction was not required for the loss of C(2)H(2) to occur. Enrichment cultures were obtained which grew anaerobically at the expense of C(2)H(2). PMID- 16345715 TI - Fungicide Enhancement of Nitrogen Fixation and Colonization of Phaseolus vulgaris by Rhizobium phaseoli. AB - The number and weight of pods and the weight and nitrogen content of the tops of beans (Phaseolus vulgaris) derived from seeds inoculated with a thiram-resistant strain of Rhizobium phaseoli were increased if the seeds were treated with thiram before sowing in soil. A greater percentage of the nodules on 21-day-old plants were derived from the resistant strain, more nodules were formed, and these nodules were more effective in the presence of the fungicide than in its absence. These differences in nodule numbers were no longer present in 56-day-old plants, and only a small percentage of the nodules contained the resistant strain. The abundance of the fungicide-tolerant R. phaseoli increased rapidly soon after planting the seed and subsequently fell markedly, but the rate of decline was less if the seeds had been treated with the chemical. Protozoa also proliferated if thiram had not been applied to the seed, but their numbers were deleteriously influenced by thiram. Bdellovibrio, bacteriophages, and lytic micro-organisms acting on R. phaseoli were rare under these conditions. Ciliates and flagellated protozoa were initially suppressed by planting thiram-coated bean seeds in nonsterile soil, but the former were inhibited longer than the latter and the ciliate numbers never fully recovered if the seeds were treated with the fungicide. The resistant strain grew well in sterile soil also inoculated with a protozoa-free mixture of soil microorganisms whether thiram was added or not, but after an initial rise in numbers, its abundance fell if the mixture contained protozoa; the rate of this fall was delayed by the fungicide. The numbers of R. phaseoli were consistently less in sterile soil inoculated with the rhizobium plus a mixture of soil microorganisms containing ciliates and other protozoa than if the inoculum contained other protozoa but no ciliates. These results suggest that a suppression of protozoa, and possibly especially the ciliates, accounts for the enhanced growth of beans and the greater initial frequency of nodules formed by the thiram-resistant R. phaseoli in the presence of this fungicide. Thiram applied to uninoculated seed enhanced bean growth if thiram-resistant R. phaseoli were present in soil. PMID- 16345716 TI - Denitrification and nitrogen fixation in alaskan continental shelf sediments. AB - Rates of nitrogen fixation and denitrification were measured in Alaskan continental shelf sediments. In some regions, rates of nitrogen fixation and denitrification appeared to be equal; in other areas, rates were significantly different. Potential rates of denitrification were found to be limited primarily by the available nitrate substrate. Major regional differences in rates of denitrification were not statistically significant, but significant differences were found for nitrogen fixation rates in different regions of the Alaskan continental shelf. Estimated net losses of nitrogen from Bering Sea sediments were calculated as 1.8 x 10 g of N/yr. Experimental exposure of continental shelf sediments to petroleum hydrocarbons reduced rates of nitrogen fixation and denitrification in some cases but not others. Long-term exposure was necessary before a reduction in nitrogen fixation rates was observed; unamended rates of denitrification but not potential denitrification rates (NO(3) added) were depressed after exposure to hydrocarbons. PMID- 16345717 TI - Production of Ethanol from d-Xylose by Using d-Xylose Isomerase and Yeasts. AB - d-Xylulose, an intermediate of d-xylose catabolism, was observed to be fermentable to ethanol and carbon dioxide in a yield of greater than 80% by yeasts (including industrial bakers' yeast) under fermentative conditions. This conversion appears to be carried out by many yeasts known for d-glucose fermentation. In some yeasts, xylitol, in addition to ethanol, was produced from d-xylulose. Fermenting yeasts are also able to produce ethanol from d-xylose when d-xylose isomerizing enzyme is present. The results indicate that ethanol could be produced from d-xylose in a yield of greater than 80% by a two-step process. First, d-xylose is converted to d-xylulose by xylose isomerase. d-Xylulose is then fermented to ethanol by yeasts. PMID- 16345718 TI - Catabolism of substituted benzoic acids by streptomyces species. AB - Four thermotolerant actinomycetes from soil, identified as Streptomyces albulus 321, Streptomyces sioyaensis P5, Streptomyces viridosporus T7A, and Streptomyces sp. V7, were grown at 45 degrees C in media containing either benzoic acid or hydroxyl- and methoxyl-substituted benzoic acids as the principal carbon sources. Benzoic acid was converted to catechol; p-hydroxybenzoic, vanillic, and veratric acids were converted to protocatechuic acid; and m-hydroxybenzoic acid was converted to gentisic acid. Catechol, protocatechuic acid, and gentisic acid were cleaved by catechol 1,2-dioxygenase, protocatechuate 3,4-dioxygenase, and gentisate 1,2-dioxygenase, respectively. Dioxygenases appeared only in induced cultures. m-Hydroxybenzoic, m-anisic, and p-anisic acids were gratuitous inducers of dioxygenases in some strains. One strain converted vanillic acid to guaiacol. PMID- 16345719 TI - Tin and tin-resistant microorganisms in chesapeake bay. AB - Sediment and water samples from nine stations in Chesapeake Bay were examined for tin content and for microbial populations resistant to inorganic tin (75 mg of Sn liter as SnCl(4).5H(2)O) or to the organotin compound dimethyltin chloride [15 mg of Sn liter as (CH(3))(2)SnCl(2)]. Tin concentrations in sediments were higher (3.0 to 7.9 mg kg) at sites impacted by human activity than at open water sites (0.8 to 0.9 mg kg), and they were very high (239.6 mg kg) in Baltimore Harbor, which is impacted by both shipping and heavy industry. Inorganic tin (75 mg Sn liter) in agar medium significantly decreased viable counts, but its toxicity was markedly reduced in liquid medium; it was not toxic in medium solidified with silica gel. Addition of SnCl(4).5H(2)O to these media produced a tin precipitate which was not involved in the metal's toxicity. The data suggest that a soluble tin-agar complex which is toxic to cells is formed in agar medium. Thus, the toxicity of tin depends more on the chemical species than on the metal concentration in the medium. All sites in Chesapeake Bay contained organisms resistant to tin. The microbial flora was more sensitive to (CH(3))(2)SnCl(2) than to SnCl(4).5H(2)O. The elevated level of tin-resistant microorganisms in some aeas not containing unusually high tin concentrations suggests that factors other than tin may participate in the selection for a tin-tolerant microbial flora. PMID- 16345720 TI - Microbiological, biochemical, and electron microscopic characterization of a pectinatus strain. AB - A spoilage organism isolated from turbid beer is described. The bacterium was gram negative, catalase negative, strictly anaerobic, and rod shaped, having flagella only on one side of the cell. The main metabolic product was propionic acid. In addition acetic, succinic, and lactic acids and acetoin were formed. Malonate inhibited the production of propionic acid by the strain studied and by both Pectinatus and Propionibacterium strains. The guanine-plus-cytosine content of deoxyribonucleic acid was 36 mol%. Differences between this strain and Pectinatus strains were 2 to 5 percentage points. Immunofluorescent staining and gel diffusion precipitin tests revealed that the antigenic structure differed from those of Pectinatus strains. The isolated organism can, despite some differences, be regarded as belonging to the genus Pectinatus. PMID- 16345721 TI - Microcultural study of bacterial size changes and microcolony and ultramicrocolony formation by heterotrophic bacteria in seawater. AB - With a microculture technique and time-lapse, phase-contrast photomicrography, it was possible to follow the division of individual cells and the development of microcolonies of bacteria in freshly collected marine water samples. A certain number of marine bacteria, upon inoculation onto a nutrient rich agar surface, displayed an increase in size as well as a high growth rate. Other bacteria were identified as very small marine bacteria (ultramicrobacteria). These had a very slow growth rate when inoculated onto a nutrient-rich agar surface. These latter cells formed very small microcolonies (ultramicrocolonies), and cell size did not increase significantly. These two types of marine heterotrophs could be described in terms of zymogenous and autochthonous bacteria, a concept used by Winogradsky for describing soil microorganisms. PMID- 16345722 TI - Morphological survey of microbial mats near deep-sea thermal vents. AB - A microscopic survey is presented of the most commonly observed and morphologically conspicuous microorganisms found attached to natural surfaces or to artificial materials deposited in the immediate vicinity of thermal submarine vents at the Galapagos Rift ocean spreading zone at a depth of 2,550 meters. Of special interest were the following findings: (i) all surfaces intermittently exposed to H(2)S-containing hydrothermal fluid were covered by layers, ca. 5 to 10 mum thick, of procaryotic, gram-negative cells interspaced with amorphous metal (Mn-Fe) deposits; (ii) although some of the cells were encased by dense metal deposits, there was little apparent correlation between metal deposition and the occurrence of microbial mats, (iii) highly differentiated forms appeared to be analogues of certain cyanobacteria, (iv) isolates from massive mats of a prosthecate bacterium could be identified as Hyphomicrobium spp., (v) intracellular membrane systems similar to those found in methylotrophic and nitrifying bacteria were observed in approximately 20% of the cells composing the mats, (vi) thiosulfate enrichments made from mat material resulted in isolations of different types of sulfur-oxidizing bacteria including the obligately chemolithotrophic genus Thiomicrospira. PMID- 16345723 TI - Nitrogen Fixation by the Photosynthetic Sulfur Bacterium Chlorobium phaeobacteroides from Lake Kinneret. AB - N(2) fixation by Chlorobium phaeobacteroides from Lake Kinneret was dependent on ammonia concentration and light intensity. In the thermocline of Lake Kinneret, N(2) fixation and photosynthesis were low. It was concluded that the bacteria do not contribute significantly to the organic nitrogen load of the lake. PMID- 16345724 TI - Poliovirus Retention in 75-cm Soil Cores After Sewage and Rainwater Application. AB - [This corrects the article on p. 1033 in vol. 40.]. PMID- 16345725 TI - Analysis, Production, and Isolation of an Extracellular Laccase from Polyporus anceps. AB - [This corrects the article on p. 1003 in vol. 40.]. PMID- 16345726 TI - Pectinase Activity of Anaerobic and Facultatively Anaerobic Bacteria Associated with Soft Rot of Yam (Diascorea rotundata). AB - Anaerobic and facultatively anaerobic bacteria associated with soft rot of yam (Diascorea rotundata) were isolated by the looping-out method and found to consist of Clostridium (three isolates), Corynebacterium (three isolates), Vibrio (one isolate), and Bacillus lentus (one isolate). Enzyme assay for hydrolase, lyase, and pectinesterase activities by the cup-plate method showed that except for Vibrio sp., B. lentus, and two isolates of Corynebacterium no pectinase activity could be detected for organisms cultured on pectin medium. Most of the cultures on yam tissue, however, showed activities for the three enzymes. The viscometric assay for hydrolase and lyase enzymes indicated a significant level of hydrolase activity (a 40.90% decrease in viscosity for Vibrio sp. and Corynebacterium spp.), but no lyase activity for most of the isolates. Two isolates of Corynebacterium and B. lentus caused changes in fresh yams suggestive of soft rot. PMID- 16345727 TI - Physiological criteria and mycotoxin production as AIDS in identification of common asymmetric penicillia. AB - The taxonomy of the asymmetric (predominantly terverticillate) penicillia is based on morphological differences that leave identification difficult. The application of physiological criteria facilitated the identification of the common asymmetric penicillia investigated. Changes in the placement of some strains of these penicillia made the connection to mycotoxin-producing ability clearer. The classical criterion of conidium color was deemphasized and replaced by the following criteria: (i) growth on nitrite-sucrose agar and (ii) growth and acid (and subsequent base) production on creatine-sucrose agar (containing bromocresol purple). Other criteria used or developed were: (iii) growth on sorbic acid plus benzoic acid agar (50 + 50 ppm, pH 3.8), (iv) growth on an agar containing 1,000 ppm propionic acid (pH 3.8), (v) growth on an agar containing 0.5% acetic acid, (vi) growth at 37 degrees C, (vii) growth rate on an agar containing 0.1% pentachloronitrobenzene, (viii) production of extracellular tricaproinase, and (ix) fasciculation on a medium containing 10 ppm botran (2,6 dichloro-4-nitroanilin). The pattern of extracellular metabolites after thin layer chromatography was used as a chemotaxonomic criterion. The species investigated, the number of isolates investigated, and the toxins which some of these isolates produce were: Penicillium roqueforti (18) (patulin), P. citrinum (11) (citrinin), P. patulum (9) (patulin and griseofulvin), P. expansum (patulin and citrinin), P. hirsutum (13), P. brevicompactum (19), and P. chrysogenum (12). Widespread species of the P. cyclopium, P. viridicatum, and P. expansum series of Raper and Thom (A Manual of the Penicillia, 1949) were subdivided into four new groups: "P. crustosum pA" (29) (penitrem A), "P. melanochlorum" (29), "P. cyclopium p" (119) (penicillic acid and infrequently penitrem A), and "P. viridicatum o-c" (43) (ochratoxin A and citrinin). "P. viridicatum o-c" was separated from "P. cyclopium p" due to its ability to grow on nitrite as sole nitrogen source. The species and groups investigated were related to the new taxonomic classification of the genus Penicillium according to Pitt. PMID- 16345728 TI - Evaluation of the total carbonaceous biochemical oxygen demand method, using a compensated recording respirometer. AB - The total carbonaceous biochemical oxygen demand test is described, and experimental data demonstrating its stoichiometry, precision, and accuracy are presented. The test is more reproducible and faster than the current 5-day biochemical oxygen demand test procedure, and if a respirometer is used, the effects of toxic chemicals, pH changes, and nutrient imbalances can be routinely monitored. The design principles of a multichannel compensated recording respirometer suitable for this test are described. PMID- 16345729 TI - Second-order model to predict microbial degradation of organic compounds in natural waters. AB - The reliability of second-order rate constants for assessing microbial degradation kinetics in natural waters was examined by using three compounds that undergo hydrolytic degradation. The butoxyethyl ester of 2,4 dichlorophenoxyacetic acid was studied in water samples from 31 sites, malathion was examined in water from 14 sites, and chlorpropham was studied in samples from 11 sites. The coefficient of variation for rate constants for each compound was less than 65% over all sites. Additional studies indicated that the rate conformed to second-order kinetics; that is, the rate was proportional to both bacterial and xenobiotic concentrations. PMID- 16345730 TI - Microbial Transformation of beta-Ionone and beta-Methylionone. AB - Aspergillus niger JTS 191 was selected from many microorganisms tested as capable of converting ionones to other compounds having aromas. The individual transformation products from beta-ionone were isolated and identified by comparison with synthetically derived compounds. The major products were (R)-4 hydroxy-beta-ionone and (S)-2-hydroxy-beta-ionone. 2-Oxo-, 4-oxo-, 3,4-dehydro-, 2,3-dehydro-4-oxo-, 3,4-dehydro-2-oxo-, (S)-2-acetoxy-, (R)-4-acetoxy-, and 5,6 epoxy-beta-ionone and 4-(2,3,6-trimethylphenyl)-but-3-en-2-one were also identified. Analogous transformation products of beta-methylionone also were identified. Based on gas-liquid chromatographic analysis during the fermentation, we propose two main oxidative pathways of beta-ionone. The results of this study suggest that these transformations of beta-ionones may be useful as tobacco flavoring compounds. PMID- 16345731 TI - Fate of the benzene ring of linear alkylbenzene sulfonate in natural waters. AB - The biodegradability of linear alkylbenzene sulfonate (LAS) was studied in water samples collected from a receiving stream at locations above and below the discharge of a municipal wastewater treatment plant. Rates of primary biodegradation were determined for a commercial LAS mixture by a modified methylene blue-active substance method. Rates of LAS ultimate degradation were determined by radiochemical methods, using a C(12) LAS homolog uniformly labeled with C in the benzene ring. The C(12) LAS was tested at low concentrations (50 and 500 mug/liter) comparable to those existing in the receiving stream. Loss of methylene blue-active substance response over time occurred rapidly in water samples containing sediment collected from below the treatment plant, with an estimated half-life for LAS of 0.23 days. Evolution of CO(2) during mineralization of the benzene ring occurred rapidly in the same samples, with a half-life for the benzene ring of 0.73 day. Mineralization of the benezene ring was also observed in river water containing no sediments and in river water and sediment samples collected from above the treatment plant. However, the rate of degradation was reduced in these cases, with half-lives for ring carbon ranging from 1.4 to 14 days. Although LAS degradation was enhanced in the presence of sediments, adsorption of LAS to the clay-silt fraction of river sediments was low, and most of the radioactivity was bound to biomass. PMID- 16345732 TI - Physiological Characteristics of Cowpea Rhizobia: Evaluation of Symbiotic Efficiency in Vigna unguiculata. AB - One fast-growing and three slow-growing strains of Rhizobium (isolated from cowpeas) were evaluated for symbiotic performance on Vigna unguiculata (L.) Walp. cultivar California no. 5 blackeyes. Plants inoculated with slow-growing strains 176A22, 176A30, and 176A32 developed a maximum acetylene reduction activity of 24.6, 27.0, and 32 mumol of ethylene formed per plant per h, respectively, versus 6.4 mumol per plant per h in plants inoculated with the fast-growing strain 176A28. When inoculated with approximately equal proportions of rhizobia, the fast-growing strain 176A28 produced 95% of the nodules when challenged with the slow-growing strain 176A22, but formed only 6% of the nodules when challenged with the slow-growing strain 176A30. Consequently, there was no relation between the growth rate in vitro and the capability of rhizobia to compete for nodule forming sites. Plants inoculated with strain 176A28 and subjected to drought during the vegetative growth period recovered to the same level of nitrogen fixation and nodulation as those that received adequate irrigation. On the other hand, plants inoculated with strains 176A22, 176A30, and 176A32 failed to achieve the same levels of nodulation and nitrogen fixation under drought as compared with irrigated conditions. PMID- 16345733 TI - Mycophenolic Acid Production by Penicillium brevicompactum on Solid Media. AB - When grown on Czapek-Dox agar, Penicillium brevicompactum produced mycophenolic acid after a vegetative mycelium had been formed and as aerial hyphae were developing. Nutrients were still plenteous in the agar when the synthesis began. If aerial hyphal development was prevented by placing a dialysis membrane over the growing fungus, no mycophenolic acid was produced. When the dialysis membrane was peeled back and, as a consequence, production of aerial hyphae began, mycophenolic acid biosynthesis was observed. We concluded that mycophenolic acid was produced only by P. brevicompactum colonies that possessed an aerial mycelium. PMID- 16345734 TI - Problematic occurrence of tyrosine crystals in the thai soybean paste tao chieo. AB - In a Bangkok soy sauce factory that had recently converted to controlled inoculum and incubation for the koji stage of fermentation, a problem arose with unsightly white particles in the soybean paste condiment called tao chieo. This problem had not arisen during the previous history of the factory where the koji stage of the fermentation was uncontrolled. Microscopic examination of the particles showed that they were crystalline. Physical separation of the crystals followed by solvent extraction, recrystallization, chemical characterization, and spectroscopy showed that they consisted of tyrosine. Tao chieo prepared by using low- or high-protease strains of Aspergillus sp. indicated that the tyrosine crystals resulted from mold proteolysis of the soybean substrate. Addition of polyethylene glycol at the beginning of the moromi fermentation reduced the severity of crystal formation. PMID- 16345735 TI - Surface changes in mild steel coupons from the action of corrosion-causing bacteria. AB - Changes which occur on the surface of mild steel coupons submerged in cultures of an Fe(III)-reducing bacterium, isolated from corroded pipe systems carrying crude oil, were studied microscopically to investigate the interaction between the corrosion-causing bacterium and the corroding mild steel coupon. Under micro aerobic conditions and in the absence of the bacteria, a dense, crystalline, amorphous coat formed on the surface of the steel coupons. In the presence of bacteria the surface coat was extensively removed, exposing the bare metal to the environment. After about 2 weeks of exposure, the removal of the surface coating was followed by colonization of the metal surface by the bacteria. Colonization was mediated by fibrous, exopolysaccharidic material formed by the bacteria. Extension of studies to other bacteria isolated from crude oil and corroded pipes reveals that the formation of exopolysaccharide fibers and possession of adherent properties are common characteristics of bacteria from crude oil systems. PMID- 16345736 TI - Fate of immediate methane precursors in low-sulfate, hot-spring algal-bacterial mats. AB - The fates of acetate and carbon dioxide were examined in several experiments designed to indicate their relative contributions to methane production at various temperatures in two low-sulfate, hot-spring algal-bacterial mats. [2 C]acetate was predominantly incorporated into cell material, although some CH(4) and CO(2) was produced. Acetate incorporation was reduced by dark incubation in short-term experiments and severely depressed by a 2-day preincubation in darkness. Autoradiograms showed that acetate was incorporated by long filaments resembling phototrophic microorganisms of the mat communities. [H]acetate was not converted to CH(4) in samples from Octopus Spring collected at the optimum temperature for methanogenesis. NaHCO(3) was readily converted to CH(4) at temperatures at which methanogenesis was active in both mats. Comparisons of the specific activities of methane and carbon dioxide suggested that of the methane produced, 80 +/- 6% in Octopus Spring and 71 +/- 21% in Wiegert Channel were derived from carbon dioxide. Addition of acetate to 1 mM did not reduce the relative importance of carbon dioxide as a methane precursor in samples from Octopus Spring. Experiments with pure cultures of Methanobacterium thermoautotrophicum suggested that the measured ratio of specific activities might underestimate the true contribution of carbon dioxide in methanogenesis. PMID- 16345737 TI - Heterotrophic potentials and hydrocarbon biodegradation potentials of sediment microorganisms within the athabasca oil sands deposit. AB - Techniques for the enumeration and the determination of the potential activity of disturbed sediment mixed populations at control sites and sites within the Athabasca oil sands formation were applied to August and December samples. These techniques included the determination of general heterotrophic potential for the assimilation and respiration of glutamate, which indicated no oil sand-related changes in the sediments but which indicated a significant seasonal change. Enumeration by epifluorescence direct counts, oil sand hydrocarbon plate counts, and most-probable-number determinations of [C]hexadecane and [C]-naphthalene degraders indicated that only the plate count was sensitive to increased numbers of oil sand-related hydrocarbon-oxidizing microorganisms within the oil sands deposit. Unlike the most probable number determinations of [C]hexadecane and [C]naphthalene degraders, however, the biodegradation potential results of these substrates indicated a significant increase in activity at oil sands sites. These biodegradation potentials also showed a marked seasonal fluctuation. Although the biodegradation potentials and the endogenous hydrocarbon plate counts indicated an oil sand-adapted mixed sediment population, the results of these techniques did not correlate well with the concentrations of bituminous hydrocarbons in the sediments. The results suggest that a general capability for hydrocarbon oxidation exists in the Athabasca River system and that this capability is enhanced within the natural bounds of the Athabasca oil sands. PMID- 16345738 TI - In vitro microbial degradation of bituminous hydrocarbons and in situ colonization of bitumen surfaces within the athabasca oil sands deposit. AB - Bituminous hydrocarbons extracted from the Athabasca oil sands of north-eastern Alberta were adsorbed onto filter supports and placed at sites in the Athabasca River and its tributaries where these rivers come in contact with the oil sands formation. Colonization of the hydrocarbon surfaces at summer and winter ambient temperatures was examined by scanning and transmission electron microscopy as well as by epifluorescence microscopy of acridine orange-stained cross sections. Ruthenium red and alkaline bismuth stains visualized an association of bacteria with the hydrocarbon surface which was mediated by bacterial polysaccharides. Bacteria apparently lacking a glycocalyx were also found closely associated with the surface of the hydrophobic substrate and in channels within the substrate. A solvent precipitation and column chromatographic fractionation of the bitumen was followed by cross-tests for growth on the fractions by various isolated sediment microorganisms, as determined by epifluorescence count. All fractions except the asphaltenes supported the growth of at least two of the isolates, although fractionation of degraded bitumen revealed that the saturate, aromatic, and first polar fractions were preferentially degraded. PMID- 16345739 TI - Calcium-Induced Alteration of Cellular Morphology Affecting the Resistance of Lactobacillus acidophilus to Freezing. AB - Examination of factors affecting the resistance of Lactobacillus acidophilus NCFM culture concentrates to freeze injury induced during frozen storage at -20 degrees C revealed that calcium supplementation of the growth medium contributed to the storage stability of cells prepared in static culture. Culture concentrates of L. acidophilus NCFM were prepared from cells propagated in MRS broth or MRS broth supplemented with 0.1% calcium carbonate, calcium chloride, or calcium phosphate. After 28 days of frozen storage at -20 degrees C, concentrated cells (3.2 x 10 colony-forming units per ml) prepared from MRS broth cultures showed an 84% reduction in viable cells. Of the remaining viable cells, 88% were sublethally injured and unable to form colonies on MRS agar supplemented with 0.15% bile. Cells prepared in calcium-supplemented MRS broths demonstrated more resistance to frozen storage. Viability and injury losses in the frozen concentrates were limited to 10 to 39% and 3 to 23%, respectively. It was observed that calcium supplementation of MRS medium resulted in a morphological transition of L. acidophilus NCFM from filamentous to bacilloid rods, and the bacilloid cells were more resistant to freezing and storage at conventional freezer temperatures. The results suggest that the morphology of the L. acidophilus cell may be an important consideration in the preparation of freeze stable culture concentrates. PMID- 16345740 TI - Volatile Compounds Associated with Spoilage of Vacuum-Packaged Sliced Luncheon Meat by Brochothrix thermosphacta. AB - By using gas chromatography-mass spectrometry, seven volatile compounds were identified in vacuum-packaged sliced corned beef spoiled by Brochothrix thermosphacta under aerobic conditions. Acetoin and diacetyl appeared to be of major sensory significance. PMID- 16345741 TI - Inhibitory effect of nitrapyrin on three genera of ammonia-oxidizing nitrifiers. AB - Five strains of Nitrosomonas and one each of Nitrosospira and Nitrosolobus were examined for sensitivity to the nitrification inhibitor nitrapyrin. Considerable variation in sensitivity was observed, with some strains about five times more resistant than others. Sensitivity to nitrapyrin varied more with strain than with genus. PMID- 16345742 TI - Technique for simultaneous determination of [s]sulfide and [C]carbon dioxide in anaerobic aqueous samples. AB - A technique for the simultaneous determination of [S]sulfide and [C]carbon dioxide produced in anaerobic aqueous samples dual-labeled with [S]sulfate and a C-organic substrate is described. The method involves the passive distillation of sulfide and carbon dioxide from an acidified water sample and their subsequent separation by selective chemical absorption. The recovery of sulfide was 93% for amounts ranging from 0.35 to 50 mumol; recovery of carbon dioxide was 99% in amounts up to 20 mumol. Within these delineated ranges of total sulfide and carbon dioxide, 1 nmol of [S]sulfide and 7.5 nmol of [C]carbon dioxide were separated and quantified. Correction factors were formulated for low levels of radioisotopic cross-contamination by sulfide, carbon dioxide, and volatile organic acids. The overall standard error of the method was +/-4% for sulfide and +/-6% for carbon dioxide. PMID- 16345743 TI - Problems associated with the use of azide as an inhibitor of microbial activity in soil. AB - Problems associated with the use of sodium or potassium azide as an inhibitor of microbial activity in soil include erroneous CO(2) evolution readings, shifts in inhibitor concentration, rise in soil pH, and explosion hazard. PMID- 16345744 TI - Aberrant Forms of Streptococcus cremoris HP. AB - The cheese starter strain, Streptococcus cremoris HP, produced variant colonies when streaked on the surface of solid media and incubated at 30 or 37 degrees C or in the presence of penicillin. Serial plating and incubation at 37 degrees C or in the presence of penicillin resulted in the production of variants. Subculture followed by incubation at 25 degrees C or in the absence of penicillin resulted in the reversion or partial reversion to the parent form. Colony morphology and cell morphology exhibited the characteristics of the L-phase. Evidence suggested that the aberrant forms of S. cremoris at 30 degrees C were transitional phase variants but at 37 degrees C and in the presence of penicillin they were L-phase variants. Electron micrographs showed that the cell walls of the variant cells were defective and that there were differences in the density and the organization of the cytoplasmic constituents compared with the parent cell. PMID- 16345745 TI - Syntrophomonas wolfei gen. nov. sp. nov., an Anaerobic, Syntrophic, Fatty Acid Oxidizing Bacterium. AB - An anaerobic, nonphototrophic bacterium that beta-oxidizes saturated fatty acids (butyrate through octanoate) to acetate or acetate and propionate using protons as the electron acceptor (H(2) as electron sink product) was isolated in coculture with either a non-fatty acid-degrading, H(2)-utilizing Desulfovibrio sp. or methanogens. Three strains of the bacterium were characterized and are described as a new genus and species, Syntrophomonas wolfei. S. wolfei is a gram negative, slightly helical rod with round ends that possesses between two to eight flagella laterally inserted along the concave side of the cell. It has a multilayered cell wall of the gram-negative type. The presence of muramic acid, inhibition of growth by penicillin, and increased sensitivity of the cells to lysis after treatment with lysozyme indicate that peptidoglycan is present in the cell wall. Cells of S. wolfei contain poly-beta-hydroxybutyrate. Isoheptanoate was degraded to acetate, isovalerate, and H(2). Carbohydrates, proteinaceous materials, alcohols, or other tested organic compounds do not support growth. Common electron acceptors are not utilized with butyrate as the electron donor. Growth and degradation of fatty acids occur only in syntrophic association with H(2)-using bacteria. The most rapid generation time obtained by cocultures of S. wolfei with Desulfovibrio and Methanospirillum hungatei is 54 and 84 h, respectively. The addition of Casamino Acids but neither Trypticase nor yeast extract stimulated growth and resulted in a slight decrease in the generation time of S. wolfei cocultured with M. hungatei. The addition of H(2) to the medium stopped growth and butyrate degradation by S. wolfei. PMID- 16345746 TI - Detection of rotavirus in sewage. AB - FOR DETECTION OF ROTAVIRUS, DOMESTIC SEWAGE WAS CONCENTRATED BY TWO DIFFERENT METHODS: (i) adsorption to and elution from positively charged Seitz filters, followed by ultracentrifugation, and (ii) chemical precipitation. The concentrated fluids were tested by an enzyme-linked immunosorbent assay and electron microscopy. In 6 of 24 (25%) samples, rotavirus was detectable after the combined filtration and ultracentrifugation technique with both an enzyme-linked immunosorbent assay and electron microscopy. No positive results were obtained after chemical precipitation. PMID- 16345747 TI - Primary production and biovolume of various phototrophic plankton size fractions in three southeastern United States reservoirs. AB - Plankton size classes of <3 mum consisting largely of unicellular cyanobacteria accounted for 15 to 40% of the total primary production and generally represented <5% of the total phototrophic plankton biovolume in three South Carolina reservoirs. PMID- 16345748 TI - Chemically Defined Minimal Medium for Growth of the Anaerobic Cellulolytic Thermophile Clostridium thermocellum. AB - A minimal chemically defined medium has been developed for Clostridium thermocellum. The growth factors required are biotin, pyridoxamine, vitamin B(12), and p-aminobenzoic acid. PMID- 16345749 TI - Serological Diversity of Nitrobacter spp. from Soil and Aquatic Habitats. AB - Serotypic diversity among Nitrobacter spp. isolates is greater than previously reported. Typing with fluorescent antibodies prepared against 11 Nitrobacter spp. cultures isolated from soil and water placed the isolates into six serogroups. When these fluorescent antibodies were applied to a group of 16 additional isolates, 8 were identifiable by cross-reaction to 3 of the 11 fluorescent antibodies. Some nitrite-oxidizing enrichment cultures from different habitats contained cross-reacting strains of Nitrobacter spp. PMID- 16345750 TI - trans-Monounsaturated Acids in a Marine Bacterial Isolate. AB - A sedimentary bacterial isolate has been shown to contain trans-monounsaturated fatty acids (6% of the total fatty acids). The ratio of trans- to cis-acids in this isolate was in the range 3.2 to 7.6. The identification of trans monounsaturated acids in a marine bacterium implied that the trans-acids which have been reported in recent sediments could derive, in whole or part, from direct bacterial input. PMID- 16345751 TI - Formation and Location of 1,4-beta-Glucanases and 1,4-beta-Glucosidases from Penicillium janthinellum. AB - Formation and location of 1,4-beta-glucanases and 1,4-beta-glucosidases were studied in cultures of Penicillium janthinellum grown on Avicel, sodium carboxymethyl cellulose, cellobiose, glucose, mannose, and maltose. Endo-1,4-beta glucanases were found to be cell free, and their formation was induced by cellobiose. 1,4-beta-Glucosidases, on the other hand, were formed constitutively and were primarily cell free, but with a small amount strongly associated with the cell wall. Low 1,4-beta-glucosidase activities of periplasmic or intracellular origin were also found. A rotational viscosimetric method was developed to measure the total endo-1,4-beta-glucanase activity of the culture (broth and solids). By this method, it was possible to determine the endo-1,4 beta-glucanase activity not only in the supernatant of the culture but also on the surface of the mycelium or absorbed on residual Avicel. During a 70-liter batch cultivation of P. janthinellum, the adsorption of endo-1,4-beta-glucanases by residual and newly added 10% Avicel was measured. The adsorption of soluble protein and endo-1,4-beta-glucanases by Avicel was found to be largely independent of the pH value but dependent on temperature. PMID- 16345752 TI - Effects of pharmaceutical wastes on microbial populations in surface waters at the puerto rico dump site in the atlantic ocean. AB - A series of cruises during 1979 and 1980 to the pharmaceutical dump site located 64 km north of Arecibo, Puerto Rico, in the Atlantic Ocean, was carried out to evaluate effects of wastes on the ecology of the microflora of surface waters of the dump site. In addition to bacteriological monitoring of the waste plume created by the release of wastes from the disposal barge, stations along a series of transects, extending north from coastal waters through and beyond the dump site, were sampled. Largest numbers of culturable bacteria on marine agar were found at stations closest to shore and in the vicinity of the dump site. Bacteria recovered on marine agar were predominantly Vibrio and Aeromonas spp., with the relative abundance of these organisms decreasing as gram-positive organisms (staphylococci, micrococci, and bacilli) became dominant in areas immediately affected by waste dumping. Total numbers of bacteria (determined by acridine orange direct counts [AODC]), which were relatively stable throughout the region, and a direct estimate of viable cells (DVC), i.e., those cells responsive to additions of yeast extract and nalidixic acid, were determined by acridine orange staining and epifluorescence microscopy. Heterotrophic bacterial activity, measured by the uptake (V(max)) of C-labeled amino acids, declined relative to distance from land. Increases in specific activity indices (DVC/AODC and V(max)/AODC) were observed near the dump site. The composite results of this study, i.e., increased specific activities (determined by two methods), increased numbers of culturable marine bacteria, and marked alteration of the taxonomic composition of the culturable bacterial community in waters within and surrounding the Puerto Rico dump site, indicate demonstrable changes in the marine microbial community in the region used for waste disposal. PMID- 16345753 TI - Comparison of ethanol production by different zymomonas strains. AB - A comparison of the rates of growth and ethanol production by 11 different strains of Zymomonas revealed a wide range of characteristics, with some strains being more tolerant of high sugar or ethanol concentrations and high incubation temperatures than others. Some strains were unable to utilize sucrose; others produced large amounts of levan, and one strain grew well but produced no levan. One strain, CP4, was considerably better in all respects than most of the other strains and was chosen as a starting strain for genetic improvement of ethanol production. PMID- 16345754 TI - Enzyme thermistor analysis of penicillin in standard solutions and in fermentation broth. AB - Immobilized penicillinase was applied in an enzyme thermistor for calorimetric analysis of samples containing penicillin G. Standard solutions as well as extracts from fermentation broth were analyzed. The enzyme was applied bound either to porous glass or, when dealing with crude preparations, to the inner surface of nylon tubing. In the fermentation system studied, high concentrations of penicillin were present, thus allowing dilution to reduce the influence of the composition of the medium on the analysis. The useful linear concentration range was from 0.1 to 100 mM. The coefficient of correlation between analytical results obtained with the present method and those from conventional assays was 0.997. PMID- 16345755 TI - Separation and Some Properties of Two Intracellular beta-Glucosidases of Sporotrichum (Chrysosporium) thermophile. AB - Intracellular, inducible beta-glucosidase from the cellulolytic fungus Sporotrichum (Chrysosporium) thermophile (ATCC 42464) was fractionated by gel chromatography or isoelectric focusing into components A and B. Enzyme A (molecular weight 440,000) had only aryl-beta-glucosidase activity, whereas enzyme B (molecular weight 40,000) hydrolyzed several beta-glucosides but had only low activity against o-nitrophenyl-beta-d-glucopyranoside (ONPG). Both enzymes had temperature optima of about 50 degrees C. The pH optimum was 5.6 for enzyme A and 6.3 for enzyme B, respectively. The K(m) (ONPG) value for enzyme A was 0.5 mM, and the corresponding values for enzyme B were 0.18 mM (ONPG) and 0.28 mM (cellobiose). Enzyme B, when tested with ONPG, showed substrate inhibition at a substrate concentration above 0.4 mM which could be released by cellobiitol and other alditols. Enzyme A was isoelectric at pH 4.48, and enzyme B was isoelectric at pH 4.64. Several inhibitors were tested for their action on the activity of enzymes A and B. Both enzymes were found to be concomitantly induced in cultures with either cellobiose or cellulose as carbon source. PMID- 16345756 TI - Sensitive and accurate methodology for measuring the kinetics of concentration dependent hydrocarbon metabolism rates in seawater by microbial communities. AB - A method having sufficient sensitivity to resolve the kinetic constants for dissolved nonpolar substrate metabolism, together with the related rate constants in natural waters, is presented. The method is based on the rate of CO(2) recovery from radioactive dissolved substrate. Sensitivity is enhanced by using large seawater volumes, high-specific-activity isotopes, and by reducing background radioactivity. Before use, commercial isotopes are purified by mild alkaline hydrolysis followed by sublimation from base to remove CO(2) as well as interfering polar C-substrates. During sample analysis, chilled Tenax resin is used to remove volatile C-substrate from the nitrogen stream containing CO(2) recovered from substrate oxidation. Chromatographic evidence of purity, shown to be insufficient, is augmented by kinetic data from toluene utilization by mixed cultures and by rates in induced versus noninduced pure cultures. Accuracy is enhanced by using short (<10 h) incubation times and small hydrocarbon concentrations so that the metabolism rates in unamended natural water systems can be evaluated. Toluene metabolism rates in seawater as low as 1 pg/liter per h and at concentrations as low as 20 ng/liter have been determined. PMID- 16345757 TI - Metabolism of 4-Chloronitrobenzene by the Yeast Rhodosporidium sp. AB - The yeast Rhodosporidium sp. metabolized 4-chloronitrobenzene by a reductive pathway to give 4-chloroacetanilide and 4-chloro-2-hydroxyacetanilide as the major final metabolites. The intermediate production of 4-chloronitrosobenzene, 4 chlorophenylhydroxylamine, and 4-chloroaniline was demonstrated by high-pressure liquid chromatography. Additional studies with selected metabolites established that the metabolite 4-chloro-2-hydroxyacetanilide was produced by an initial Bamberger rearrangement of the hydroxylamine metabolite, followed by acetylation. Direct C hydroxylation of the aromatic ring was not observed in this species. No hydroxamic acid production was detected, even though significant concentrations of the nitroso and hydroxylamine precursors to this functional group were observed. PMID- 16345758 TI - Characterization of Extracellular beta-d-Galactosidase from Fusarium moniliforme Grown in Whey. AB - Extracellular lactase (beta-d-galactosidase, EC 3.2.1.23) was prepared as an ethanol precipitate from a culture of Fusarium moniliforme grown on whey. The enzyme functioned optimally at pH 3.8 to 5.0 and at 50 to 60 degrees C on both o nitrophenyl-beta-d-galactopyranoside (ONPG) and lactose. The activation energy of the enzymic hydrolysis of ONPG and lactose in the range of 20 to 55 degrees C was 8,500 and 7,200 cal (ca. 3.57 x 10 and 3.02 x 10 J)/mol, respectively. The K(m) values were 4.4 and 12.4 mM for ONPG and lactose, respectively. At optimum pH, the enzyme lost half of its activity when it was heated at 50 degrees C for 6 h; at the same pH, the loss was only 5% when the enzyme was heated at 37 degrees C for 6 h. At optimum conditions, 50% of the lactose in whey was hydrolyzed by 10 U of this enzyme in 50 h. PMID- 16345760 TI - Curvularia lunata, a New Source of Cytochalasin B. AB - A biologically active metabolite was found in crude extracts of Curvularia lunata (Wakker) Boedijn (ATCC 34690) isolated from decayed tissues of litchi fruit (Litchi chinensis Sonn.). The fungus was grown on a shredded wheat-yeast extract sucrose medium, and cultures were extracted with chloroform after 3 weeks of growth at 21 degrees C. Chloroform extracts were toxic to day-old cockerels and caused abnormal distortion of wheat coleoptile segments. The major toxin had a 50% lethal dose of 700 mg/kg (oral dose) and was a colorless crystalline material with a melting point of 218 degrees C. Elemental and high-resolution mass spectral analyses indicated a formula of C(29)H(37)NO(5) and a molecular weight of 479.62. The crystalline preparation was identified as 97% cytochalasin B and 3% cytochalasin A. The yield of cytochalasin B from C. lunata cultures grown on 4.5 kg of shredded wheat and 9 liters of yeast extract-sucrose medium was 6.24 g of purified material. PMID- 16345759 TI - R-Plasmid Transfer to and from Escherichia coli Strains Isolated from Human Fecal Samples. AB - Strains of Escherichia coli recently isolated from human feces were examined for the frequency with which they accept an R factor (R1) from a derepressed fi strain of E. coli K-12 and transfer it to fecal and laboratory strains. Colicins produced by some of the isolates rapidly killed the other half of the mating pair; therefore, conjugation was conducted by a membrane filtration procedure whereby this effect was minimized. The majority of fecal E. coli isolates accepted the R factor at lower frequencies than K-12 F, varying from 10 per donor cell to undetectable levels. The frequencies with which certain fecal recipients received the R-plasmid were increased when its R transconjugant was either cured of the R1-plasmid and remated with the fi strain or backcrossed into the parental strain. The former suggests the loss of an incompatibility plasmid, and the latter suggests the modification of the R1-plasmid deoxyribonucleic acid (DNA). In general, the fecal RE. coli transconjugants were less effective donors for K 12 F and heterologous fecal strains than was the fi K-12 strain, whereas the single strain of Citrobacter freundii examined was generally more competent. Passage of the R1-plasmid to strains of salmonellae reached mating frequencies of 10 per donor cell when the recipient was a Salmonella typhi previously cured of its resident R-plasmid. However, two recently isolated strains of Salmonella accepted the R1-plasmid from E. coli K-12 R or the RE. coli transconjugants at frequencies of 5 x 10 or less. PMID- 16345761 TI - Influence of Nitrogen Source, Thiamine, and Light on Biosynthesis of Abscisic Acid by Cercospora rosicola Passerini. AB - Abscisic acid production by Cercospora rosicola Passerini in liquid shake culture was measured with different amino acids in combination and singly as nitrogen sources and with different amounts of thiamine in the media. Production of abscisic acid was highest with aspartic acid-glutamic acid and aspartic acid glutamic acid-serine mixtures as nitrogen sources. Single amino acids that supported the highest production of abscisic acid were asparagine and monosodium glutamate. Thiamine was important for abscisic acid production. Leucine inhibited abscisic acid production. C. rosicola produced abscisic acid in the dark, but production more than doubled in the presence of light. PMID- 16345762 TI - Isolation of Streptomyces rimosus Mutants with Reduced Actinophage Susceptibility. AB - The infection of Streptomyces rimosus by the virulent actinophage RP1 was partially characterized. RP1 infection of the host cells results in a dramatic decrease in viable cell count, followed by reduced antibiotic production. Phage resistant mutants were isolated after mutagenic treatment and RP1 selective pressure. Characterization of the isolated mutants has revealed that RP1 infection had no influence on their growth and antibiotic production. However, multiplication of the phage particles in the lawns of resistant mutants was detected. Since these strains differ from the wild type in RP1 relative efficiency of plating, plaque morphology, and the time necessary for plaque appearance, they are considered to be semiresistant mutants. The propagation of RP1 on semiresistant strains is characterized by lower adsorption of phage particles and longer latent and rise periods. As a consequence, the multiplication of the phage is slower than that of their host, which consequently reduces the ratio of phage to its host, thus diluting out the phage. PMID- 16345763 TI - Derivation of Aromatic Amino Acid Mutants from a Methanol-Utilizing Yeast, Hansenula polymorpha. AB - Three classes of mutants, deregulated to enhance the flow of aromatic intermediates through the tryptophan biosynthetic branch, were obtained. 5 Fluorotryptophan, an antimetabolite of tryptophan, was employed to obtain one class of deregulated mutants. By sequential resistance development, three resistant mutants were isolated. Hansenula polymorpha strains showed greater sensitivity to 5-fluorotryptophan when growing on methanol than when growing on glucose. Yeast extract stimulated the production of total indole metabolites (indoles) by wild-type and mutant strains, with each 5-fluorotryptophan mutant producing higher amounts of these metabolites than its predecessor. Two other mutant classes were isolated: (i) a mutant resistant to anthranilate (an inhibitory intermediate in the tryptophan biosynthetic branch) and (ii) a phenylalanine-plus-tyrosine bradytroph. Each of these produced a higher extracellular titer of total indoles than its immediate parent. With respect to the overproduction of indoles, resistance to 5-fluorotryptophan was a more useful selection method than were anthranilate resistance and phenylalanine-plus tyrosine bradytrophy. PMID- 16345764 TI - Microplate technique for determining accumulation of metals by algae. AB - A microplate technique was developed to determine the conditions under which pure cultures of algae removed heavy metals from aqueous solutions. Variables investigated included algal species and strain, culture age (11 and 44 days), metal (mercury, lead, cadmium, and zinc), pH, effects of different buffer solutions, and time of exposure. Plastic, U-bottomed microtiter plates were used in conjunction with heavy metal radionuclides to determine concentration factors for metal-alga combinations. The technique developed was rapid, statistically reliable, and economical of materials and cells. Results (expressed as concentration factors) were in reasonably good agreement with literature values. All species of algae studied removed mercury from solution. Green algae proved better at accumulating cadmium than did blue-green algae. No alga studied removed zinc, perhaps because cells were maintained in the dark during the labeling period. Chlamydomonas sp. proved superior in ability to remove lead from solution. PMID- 16345765 TI - Survival of microorganisms in a rock bed under conditions simulating solar heat storage. AB - A laboratory-scale unit containing about 360 kg of washed river gravel was designed to [ill] the use of rocks for heat storage. The unit was operated under varying conditions of temperature, relative humidity, and the addition of volatile nutrients over a 4-month period. Effluent air and rock surfaces were monitored for the presence of microorganisms. After 2 weeks, virtually no microorganisms were detected in the effluent air except when dry soil or compost was added as the inoculum. A small number of heat-resistant bacteria, but no fungi, were found to survive on the rock surfaces. Microorganisms isolated were either sporeforming bacteria or actinomycetes closely resembling Thermoactinomyces vulgaris. Microbial colonization of rock beds used for solar heat storage does not appear likely under routine operation. PMID- 16345766 TI - Polyguaiacol: a useful model polymer for lignin biodegradation research. AB - A polymer of ring-labeled [C]o-methoxyphenol ([C]guaiacol) was prepared by peroxidase-H(2)O(2)-catalyzed oxidation of the C-labeled monomeric compound. The ring-labeled [C]polyguaiacol contained 67.71% carbon, 5.09% hydrogen, 27.49% oxygen, 25.44% methoxyl, and 8.60% phenolic hydroxyl. The polymer had an average molecular weight of between 5,000 and 15,000, as determined by gel chromatography. A schematic representation of the polymer, similar to previously published structures of polyguaiacols, was devised to meet these and other analytical parameters. The polymer is primarily composed of o-o and p-p-linked guaiacol moieties, with an occasional o-p-biphenyl link and some p-diphenoquinone structures. An approximate molecular formula is [C(49)O(14)H(31)](n), where n =/> 5.8. Its C(6) formula is C(6)H(2.3)O(0.3) (OH)(0.7)(OCH(3))(1.0). Polyguaiacol has many of the characteristics of a synthetic lignin. It is easier and less expensive to prepare than standard synthetic lignins (dehydrogenation polymers of coniferyl alcohol). It is degraded ([C]polyguaiacol --> CO(2)) by the lignolytic system of the white-rot fungus Phanaerochaete chrysosporium. It is suggested that [C]polyguaiacol may be of value as a substrate for lignin biodegradation research. PMID- 16345767 TI - Comparison of two direct-count methods for determining metabolizing bacteria in freshwater. AB - Planktonic bacteria collected from several freshwater environments and cultured bacteria were used to compare two methods for determining the numbers of metabolizing bacteria. The methods used were (i) reduction of 2-(rho-iodophenyl) 3-(rho-nitrophenyl)-5-phenyl tetrazolium chloride 2-(rho-iodophenyl)-3-(rho nitrophenyl)-5-phenyl to tetrazolium chloride-formazan and (ii) elongation of cells by using yeast extract and nalidixic acid. No statistically significant difference was found between methods in determining metabolizing bacteria, although significant differences (P < 0.05) were found when comparing numbers of total bacteria. A combination of the two methods yielded significant changes, both positive and negative, in the numbers of metabolizing bacteria. PMID- 16345768 TI - Physical Factors That Affect In Vitro Autographa californica Nuclear Polyhedrosis Virus Infection. AB - Of the physical parameters tested for in vitro baculovirus infection, multiplicity of infection was most important in governing percent cell infection. Most plaques formed within the first 5 min of incubation. Efficiency of infection, however, was low, and the virus titer did not diminish during prolonged incubation. Efficiency of infection improved markedly when cells or virus were preincubated with selected polyanions and polycations. Precise regulation of the pH, osmotic pressure, and ionic composition of the cell culture medium also promoted maximum in vivo infection. PMID- 16345769 TI - Degradation of phthalic acids by denitrifying, mixed cultures of bacteria. AB - Mixed cultures of bacteria, enriched from aquatic sediments, grew anaerobically on all three isomers of phthalic acid. Each culture grew anaerobically on only one isomer and also grew aerobically on the same isomer. Pure cultures were isolated from the phthalic acid (o-phthalic acid) and isophthalic acid (m phthalic acid) enrichments that grew aerobically on phthalic and isophthalic acids. Cell suspension experiments indicated that protocatechuate is an intermediate of aerobic catabolism. Pure cultures which grew aerobically on terephthalic acid (p-phthalic acid) could not be isolated from the enrichments, and neither could pure cultures that grew anaerobically on any of the isomers. Cell suspension experiments suggested that separate pathways exist for the aerobic and anaerobic oxidation of phthalic acids. Each enrichment culture used only one phthalic acid isomer under anaerobic conditions, but all isomers were simultaneously adapted for the anaerobic catabolism of benzoate. Cells grown anaerobically on a phthalic acid immediately attacked the isomer under anaerobic conditions, whereas there was a lag before aerobic breakdown occurred, and, for phthalic and terephthalic acids, chloramphenicol stopped aerobic adaptation but had no effect on anaerobic catabolism. This work suggests that phthalic acids are biodegradable in anaerobic environments. PMID- 16345770 TI - Carbon monoxide metabolism in roadside soils. AB - Air-dried soils which were equilibrated under relative humidities greater than 93% or moistened with liquid water showed marked increases in their capacities to oxidize CO to CO(2). Liquid water addition in excess of saturation resulted in lower CO oxidation rates, reflecting the limited diffusion of CO through the aqueous phase. After 35 days' storage under 100% relative humidity, the capacity for CO oxidation decreased to 21% of the value observed with a freshly collected sample. Incubation of this stored soil under an atmosphere containing 200 ppm of CO (250 mg/m) for 21 days resulted in a sevenfold increase in CO oxidation. A correlation was noted between the CO oxidative activity and the history of previous exposure of soils to high ambient levels of CO. The organisms responsible for CO oxidation apparently comprise a small fraction of the microbial population in the soils. With a roadside soil the oxidation of CO provided the driving force for the assimilation of CO(2). The stoichiometry of the oxidative and assimilatory reactions in soil was in the range of values reported from laboratory studies with CO chemoautotrophs (carboxydobacteria). It is proposed that the population and activity of CO-oxidizing microorganisms increase in response to increasing levels of CO in the environment. PMID- 16345771 TI - Methanol Inhibition in Continuous Culture of Hansenula polymorpha. AB - Growth inhibition of Hansenula polymorpha DL-1 by methanol, formaldehyde, formate, and formic acid was examined to determine the causes of unstable behavior observed during continuous cultures on methanol. The much greater inhibition of growth by formic acid than by formate and the effect of formic acid excretion and assimilation on pH helped to explain culture dynamics observed after transitory oxygen limitations. Oxygen limitation caused by temporary reduction of agitation in a continuous fermentation caused methanol to accumulate to inhibitory concentrations. Immediately after resumption of agitation, formic acid was produced and caused culture inhibition. To ensure the stability of H. polymorpha in continuous culture, it was therefore necessary to prevent transient methanol accumulation. PMID- 16345772 TI - Fermentative conversion of cellulose to acetic Acid and cellulolytic enzyme production by a bacterial mixed culture obtained from sewage sludge. AB - A simple procedure that uses a cellulose-enriched culture started from sewage sludge was developed for producing cellulolytic enzymes and converting cellulose to acetic acid rather than CH(4) and CO(2). In this procedure, the culture which converts cellulose to CH(4) and CO(2) was mixed with a synthetic medium and cellulose and heated to 80 degrees C for 15 min before incubation. The end products formed were acetic acid, propionic acid, CO(2), and traces of ethanol and H(2). Supernatants from 6- to 10-day-old cultures contained 16 to 36 mM acetic acid. Cellulolytic enzymes in the supernatant were stable at 2 degrees C under aerobic conditions for up to 4 weeks and had the ability to hydrolyze carboxymethyl cellulose, a microcystalline cellulose, cellobiose, xylan, and filter paper to reducing sugars. PMID- 16345773 TI - Fungi and Bacteria in or on Leaves of Eelgrass (Zostera marina L.) from Chesapeake Bay. AB - Samples of green and brown leaves of eelgrass (Zostera marina L.) were incubated in seawater without an additional carbon source. Parallel leaf samples were used for acridine orange bacterial counting and water-soluble aniline blue estimation of fungal biovolume. The incubations produced no evidence that there is an eelgrass counterpart for the chytridialean symbiont which is very common in turtlegrass (Thalassia testudinum Konig). Sterile mycelium (i.e., living mycelium without identifiable propagules) was the most prevalent fungal form on incubated samples from submerged sites, whereas Dendryphiella salina and Sigmoidea sp. (marina?) were prevalent on brown leaves from the wrack line. Attempts to assay fungal biovolume in field samples indicated that the sterile mycelium observed after incubation represented the outgrowth of formerly dormant propagules or weakly established microcolonies. It was calculated that fungal biomass could not account for more than 0.5% of leaf mass, and it was probably much smaller than this, for no fungal structures were observed even in concentrated leaf homogenates. Bacterial densities fell within the range reported for other particulate substrates. A speculative estimate of bacterial productivity was 1.4x the standing stock per day. PMID- 16345774 TI - Reduction of sulfur compounds in the sediments of a eutrophic lake basin. AB - Concentrations of various sulfur compounds (SO(4), H(2)S, S, acid-volatile sulfide, and total sulfur) were determined in the profundal sediments and overlying water column of a shallow eutrophic lake. Low concentrations of sulfate relative to those of acid-volatile sulfide and total sulfur and a decrease in total sulfur with sediment depth implied that the contribution of dissimilatory sulfur reduction to H(2)S production was relatively minor. Addition of 1.0 mM Na(2)SO(4) to upper sediments in laboratory experiments resulted in the production of H(2)S with no apparent lag. Kinetic experiments with S demonstrated an apparent K(m) of 0.068 mmol of SO(4) reduced per liter of sediment per day, whereas tracer experiments with S indicated an average turnover time of the sediment sulfate pool of 1.5 h. Total sulfate reduction in a sediment depth profile to 15 cm was 15.3 mmol of sulfate reduced per m per day, which corresponds to a mineralization of 30% of the particulate organic matter entering the sediment. Reduction of S occurred at a slower rate. These results demonstrated that high rates of sulfate reduction occur in these sediments despite low concentrations of oxidized inorganic compounds and that this reduction can be important in the anaerobic mineralization of organic carbon. PMID- 16345775 TI - Activation of Polynuclear Aromatic Hydrocarbons to Mutagens by the Marine Ciliate Parauronema acutum. AB - The marine ciliate Parauronema acutum converted 2-aminofluorene and 2 acetylaminofluorene to compounds with mutagenic activity in the Ames Salmonella test. The ciliate, however, did not activate benzo (alpha)pyrene or benzanthracene or destroy the mutagenic properties of nitrosoguanidine. Homogenates, when substituted for the liver S-9 fraction in the Salmonella/microsome test, activated 2-aminofluorene and 2-acetylaminofluorene to mutagens. Benzo(alpha)pyrene and benzanthracene were not activated, nor was nitrosoguanidine inactivated. Phenobarbitol did not induce or increase the amount of activating activity. The activation showed no requirement for the reduced nicotinamide adenine dinucleotide phosphate-regenerating system required by liver P-450 cytochromes. Upon differential sedimentation of a cell homogenate, the majority of the activity sedimented with a small-particle fraction with sedimentation properties like those of microsomes from higher eucaryotes. Benzo(alpha)pyrene, although not metabolized, was accumulated by cultures of P. acutum at a linear rate and was not appreciably released (10%) after removal of benzo(alpha)pyrene from the incubation medium. Hence, this ciliate could convert certain polynuclear aromatic hydrocarbons to mutagens and accumulate others. PMID- 16345776 TI - Microbiological Systems in Organic Synthesis: Preparative-Scale Resolution of (RS)-Glaucine by Fusarium solani and Stereospecific Oxidation of (R)-(-)-Glaucine by Aspergillus flavipes. AB - The destructive resolution of (6aR,S)-glaucine (Ic) was accomplished by oxidation of the (6aS)-(+)-enantiomer (Ia), using Fusarium solani ATCC 12823 to yield the unnatural alkaloid (6aR)-(-)-glaucine (Ib). Eighteen cultures were examined for their ability to metabolize the (6aR)-(-)-enantiomer (Ib), and Aspergillus flavipes ATCC 1030 was found to catalyze the stereoselective oxidation of this substrate to didehydroglaucine. Thus, it has been demonstrated that "R" and "S" organisms exist with regard to the oxidation of aporphines to didehydroaporphines. PMID- 16345777 TI - Acidophilic, heterotrophic bacteria of acidic mine waters. AB - Obligately acidophilic, heterotrophic bacteria were isolated both from enrichment cultures developed with acidic mine water and from natural mine drainage. The bacteria were grouped by the ability to utilize a number of organic acids as sole carbon sources. None of the strains were capable of chemolithotrophic growth on inorganic reduced iron and sulfur compounds. All bacteria were rod shaped, gram negative, nonencapsulated, motile, capable of growth at pH 2.6 but not at pH 6.0, catalase and oxidase positive, strictly aerobic, and capable of growth on citric acid. The bacteria were cultivatable on solid nutrient media only if agarose was employed as the hardening agent. Bacterial densities in natural mine waters ranged from approximately 20 to 250 cells per ml, depending upon source and culture medium. Ferric hydrates and stream vegetation contained from 1,500 to over 7 x 10 cells per g. PMID- 16345778 TI - Effect of silicate grain shape, structure, and location on the biomass and community structure of colonizing marine microbiota. AB - Microbiota colonizing silica grains of the same size and water pore space, but with a different microtopography, showed differences in biomass and community structure after 8 weeks of exposure to running seawater. The absence of surface cracks and crevices resulted in a marked diminution of the total microbial biomass measured as lipid phosphate and total extractable palmitic acid. With increasing smoothness of the sand grain surface, examination of the community structure showed a marked decrease in procaryotes and algal microeucaryotes, with a relative increase in microeucaryotic grazers. A comparison of the colonizing sediment incubated in running seawater or at 32 m on the sea floor with a sediment core showed a decreased bacterial biomass with a different community structure and a decreased total microeucaryotic population of both grazers and algae. The quantitative differences in microbial biomass and community structure between the microcosms and the actual benthic population in the core were determined. PMID- 16345779 TI - Fungal metabolite from members of the genus rhizopus. AB - A fluorescent metabolite present in seven members of the genus Rhizopus was isolated. This compound appeared green before spray treatment and purple after spray treatment with p-anisaldehyde in visible light. Subsequent purification and structural elucidation of the isolated compound yielded 1-[2,6,10,14-tetramethyl 17-carbomethyl heptadecyl]-1-[2,6,10,14-tetramethyl-17-methanoyl heptadecyl] benzene. PMID- 16345780 TI - Morphological survey of microbial mats near deep-sea thermal vents. AB - [This corrects the article on p. 531 in vol. 41.]. PMID- 16345781 TI - Pectinase Activity of Anaerobic and Facultatively Anaerobic Bacteria Associated with Soft Rot of Yam (Diascorea rotundata). AB - [This corrects the article on p. 564 in vol. 41.]. PMID- 16345782 TI - Induction, Isolation, and Characterization of Aspergillus niger Mutant Strains Producing Elevated Levels of beta-Galactosidase. AB - [This corrects the article on p. 594 in vol. 41.]. PMID- 16345783 TI - Carbohydrate Fermentation by Streptococcus cremoris and Streptococcus lactis Growing in Agar Gels. AB - When lactic streptococci were embedded in agar gels and incubated at 30 degrees C, the end products of carbohydrate fermentation depended on the initial cell density, which determined the subsequent distribution and size of colonies in the gel. With high initial cell densities, microcolonies formed close together and lactose and glucose were converted almost entirely to lactate. However, inoculation with a small number of cells, which then grew to form widely spaced and comparatively large colonies, resulted in up to 30% diversion of end product, usually to formate, ethanol, and acetate. In these "low-colony-density" gel cultures, the initial rate of fermentation was exponential and only lactate was formed. However, this rate then became linear and fermentation became progressively more heterolactic. Streptococcus lactis ML(8) was the only strain among the 10 tested which remained homolactic. Incubation at temperatures either above or below the optimum for growth and metabolism decreased the diversion to end products other than lactate. The change from homo- to heterolactic fermentation appears to be caused by carbohydrate depletion in the vicinity of the colony, so that fermentation is then limited by the diffusion of substrate. Growth of cells on gel surfaces exposed to air resulted in up to 40% diversion of end product from lactate, mainly to CO(2), acetoin, 2,3-butanediol, and acetate. Six of the 12 Streptococcus cremoris strains tested remained homolactic under these aerobic conditions, whereas all 8 of the S. lactis strains tested, including ML(8), were heterolactic. PMID- 16345784 TI - Seasonal rates of methane oxidation in anoxic marine sediments. AB - Methane concentrations and rates of methane oxidation were measured in intact sediment cores from an inshore marine sediment at Jutland, Denmark. The rates of methane oxidation, determined by the appearance of CO(2) from injected CH(4), varied with sediment depth and season. Most methane oxidation was anoxic, but oxygen may have contributed to methane oxidation at the sediment surface. Cumulative rates (0- to 12-cm depth) for methane oxidation at Kysing Fjord were 3.34, 3.48, 8.60, and 17.04 mumol m day for April (4 degrees C), May (13 degrees C), July (17 degrees C), and August (21 degrees C), respectively. If all of the methane was oxidized by sulfate, it would account for only 0.01 to 0.06% of the sulfate reduction. The data indicate that methane was produced, in addition to being oxidized, in the 0- to 18-cm sediment stratum. PMID- 16345785 TI - Factors affecting oxidation of thiosalts by thiobacilli. AB - The effects of temperature, initial pH, and the concentrations of ammonium, phosphate, and heavy metals on the oxidation of thiosalts by an authentic strain of Thiobacillus thiooxidans (ATCC 8085) and by a mixed culture isolated from a base metal-processing mill effluent pond were studied. The optimum temperature was 30 degrees C and the optimum initial pH was 3.75 for both cultures using thiosulfate and for the mixed culture using tetrathionate. T. thiooxidans ATCC 8085 did not oxidize tetrathionate. For a thiosalt concentration of 2,000 ppm (2,000 mg/liter), maximal rates of destruction occurred at concentrations of ammonium ion above 2 mg/liter and in the presence of 1 mg of phosphate per liter. Under optimal conditions, the rate of thiosulfate oxidation by the pure culture was 55 +/- 3 mg/liter per h; the mixed culture oxidized thiosulfate at the rate of 40 +/- 1 mg/liter per h and tetrathionate at the rate of 50 +/- 2 mg/liter per h. Metal ions caused normal inhibition kinetics in the oxidation of thiosulfate by T. thiooxidans ATCC 8085. K(i) values were calculated for cadmium (16 mg/liter), copper (0.46 mg/liter), lead (2 mg/liter), silver (3.1 mg/liter), and zinc (33 mg/liter). Only a slight additive effect was apparent in the presence of all of these metal ions. The mixed culture of thiosalt-oxidizing bacteria was less sensitive to heavy metal inhibition; the order of inhibition of thiosulfate oxidation was Cd < Zn < Pb < Ag < Cu, and that of tetrathionate oxidation was Zn < Cd < Pb < Ag < Cu. PMID- 16345786 TI - Composting process control based on interaction between microbial heat output and temperature. AB - Rational composting process control involves the interrelated factors of heat output, temperature, ventilation, and water removal. The heat is released microbially at the expense of organic material; temperature is an effect and, because it is a determinant of microbial activity, it is also a cause of heat output; ventilation supplies oxygen and removes heat, mainly through the vaporization of water; water removal results from heat removal. These relationships were implemented in a field-scale process of static-pile configuration, using a mixture of sewage sludge and wood chips. Heat removal was matched to heat output through a temperature feedback control system, thereby maintaining biologically favorable temperatures. The observations indicate that fundamentally there are two kinds of composting systems: those that are and those that are not temperature self-limiting. The self-limiting system reaches inhibitive temperatures (>60 degrees C) which debilitate the microbial community, suppressing decomposition, heat output, and water removal. In contrast, non-self limiting temperatures (<60 degrees C) support a robust community, promoting decomposition, heat output, and water removal. PMID- 16345787 TI - Ethanol Production by Thermophilic Bacteria: Fermentation of Cellulosic Substrates by Cocultures of Clostridium thermocellum and Clostridium thermohydrosulfuricum. AB - The fermentation of various saccharides derived from cellulosic biomass to ethanol was examined in mono- and cocultures of Clostridium thermocellum strain LQRI and C. thermohydrosulfuricum strain 39E. C. thermohydrosulfuricum fermented glucose, cellobiose, and xylose, but not cellulose or xylan, and yielded ethanol/acetate ratios of >7.0. C. thermocellum fermented a variety of cellulosic substrates, glucose, and cellobiose, but not xylan or xylose, and yielded ethanol/acetate ratios of approximately 1.0. At nonlimiting cellulosic substrate concentrations ( approximately 1%), C. thermocellum cellulase hydrolysis products accumulated during monoculture fermentation of Solka Floc cellulose and included glucose, cellobiose, xylose, and xylobiose. A stable coculture that contained nearly equal numbers of C. thermocellum and C. thermohydrosulfuricum was established that fermented a variety of cellulosic substrates, and the ethanol yield observed was twofold higher than in C. thermocellum monoculture fermentations. The metabolic basis for the enhanced fermentation effectiveness of the coculture on Solka Floc cellulose included: the ability of C. thermocellum cellulase to hydrolyze alpha-cellulose and hemicellulose; the enhanced utilization of mono- and disaccharides by C. thermohydrosulfuricum; increased cellulose consumption; threefold increase in the ethanol production rate; and twofold decrease in the acetate production rate. The coculture actively fermented MN300 cellulose, Avicel, Solka Floc, SO(2)-treated wood, and steam-exploded wood. The highest ethanol yield obtained was 1.8 mol of ethanol per mol of anhydroglucose unit in MN300 cellulose. PMID- 16345788 TI - Effect of steam sterilization and gamma irradiation of peat on quality of Rhizobium inoculants. AB - Data obtained by independent tests on each of 483 batches of Rhizobium inoculants for Glycine max, Medicago sativa, and Arachis hypogaea, manufactured commercially in South Africa, are reported and discussed. Whereas the average cell count per gram per batch was well in excess of 10, inoculants for G. max and M. sativa manufactured with peat treated with gamma irradiation at a dose of 50 kGr contained significantly higher numbers of Rhizobium cells than inoculants from peat which received 25 kGr. Inoculants for M. sativa manufactured with steam sterilized peat were similar in quality to those prepared with peat irradiated at a dose of 50 kGr. Contrary to the inoculants for G. max and M. sativa, the Rhizobium strain used in inoculants for A. hypogaea was apparently insensitive to the effect on peat of the higher gamma irradiation dosage. PMID- 16345789 TI - Metabolic Activity of Fatty Acid-Oxidizing Bacteria and the Contribution of Acetate, Propionate, Butyrate, and CO(2) to Methanogenesis in Cattle Waste at 40 and 60 degrees C. AB - The quantitative contribution of fatty acids and CO(2) to methanogenesis was studied by using stirred, 3-liter bench-top digestors fed on a semicontinuous basis with cattle waste. The fermentations were carried out at 40 and 60 degrees C under identical loading conditions (6 g of volatile solids per liter of reactor volume per day, 10-day retention time). In the thermophilic digestor, acetate turnover increased from a prefeeding level of 16 muM/min to a peak (49 muM/min) 1 h after feeding and then gradually decreased. Acetate turnover in the mesophilic digestor increased from 15 to 40 muM/min. Propionate turnover ranged from 2 to 5.2 and 1.5 to 4.5 muM/min in the thermophilic and mesophilic digestors, respectively. Butyrate turnover (0.7 to 1.2 muM/min) was similar in both digestors. The proportion of CH(4) produced via the methyl group of acetate varied with time after feeding and ranged from 72 to 75% in the mesophilic digestor and 75 to 86% in the thermophilic digestor. The contribution from CO(2) reduction was 24 to 29% and 19 to 27%, respectively. Propionate and butyrate turnover accounted for 20% of the total CH(4) produced. Acetate synthesis from CO(2) was greatest shortly after feeding and was higher in the thermophilic digestor (0.5 to 2.4 muM/min) than the mesophilic digestor (0.3 to 0.5 muM/min). Counts of fatty acid-degrading bacteria were related to their turnover activity. PMID- 16345790 TI - Heat Resistance and Population Stability of Lyophilized Bacillus subtilis Spores. AB - Bacillus subtilis 5230 spores were lyophilized in 0.067 M phosphate buffer and stored at 2 to 8 degrees C for 9 to 27 months. The lyophilized spores were reconstituted with buffer or 0.9% saline, and the heat resistance was determined in a thermoresistometer. Lyophilization had no effect on the heat resistance of the spores but did result in a slight decrease in population (10 mM. Both enzymes were inhibited by Cu (>20 mM), Zn (>1.0 mM), and ethylene glycol-bis(beta-aminoethyl ether)- N,N-tetraacetic acid (>10 mM). T. reesei but not C. thermocellum cellulose-solubilizing activity was 20% inhibited by glucose (73 mM) and cellobiose (29 mM). Both cellulases preferentially cleaved the internal glycosidic bonds of cellooligosaccharides. The overall rates of cellooligosaccharide degradation were higher for T. reesei than for C. thermocellum cellulase, except that the rates of conversion of cellohexaose to cellotriose were equivalent. PMID- 16345824 TI - Release of Rhizobium spp. from Tropical Soils and Recovery for Immunofluorescence Enumeration. AB - Limitations associated with immunofluorescence enumeration of bacteria in soil derive largely from the efficiency with which cells can be separated from soil particles and collected on membrane filters for staining. Many tropical soils fix added bacteria tightly, resulting in low recoveries. Eight soils, representative of three of the major soil orders found in the tropics (oxisols, vertisols, and inceptisols), were tested for recovery of added Rhizobium strains. All except one Hawaiian andept (Typic Eutrandept) yielded recoveries ranging from <1 to 13%. Recovery from the andept was 100%. In soil-sand mixtures, addition of only a small amount of soil caused a dramatic decrease in recovery of added rhizobia. Increasing the soil content of the mixture from 0% (10 g of sand) to 50% (5 g of soil-5 g of sand) reduced recoveries from >90 to <1%. Varying the ionic strength and pH of the extracting solution did not cause marked increases in recovery. Protein solutions, ethylenediaminetetraacetate, and NaHCO(3), on the other hand, improved release of bacteria. We report a modification to the usual membrane filter immunofluorescence procedure which yielded consistently high and reproducible recovery (coefficient of variation, 30%) of rhizobia from several tropical soils. In the modified procedure, partially hydrolyzed gelatin, diluted in ammonium phosphate, was used to suspend the soil. This caused dispersion of the soil and release of the bacteria from soil flocs. The efficiency of recovery of Rhizobium spp. from several tropical and two temperate soils remained high as the content of these soils in soil-sand mixtures was increased from 0 to 100%. The modified membrane filter immunofluorescence procedure was used to follow the growth of a strain of chickpea (Cicer arietinum) Rhizobium in a sterilized oxisol. The results showed a close agreement with viable counts at different stages during the growth cycle. Diluent for the hydrolyzed gelatin also had a marked effect on recovery. The efficiency of release of Rhizobium spp. from an oxisol was in the following order for the diluents used: 0.1 M (NH(4))(2)HPO(4) > 0.1 M Na(2)HPO(4) = 0.1 M sodium-phosphate-buffered saline (pH 7.2) > 0.2 M NH(4)Cl > 0.2 KCl > NaCl = LiCl > water. PMID- 16345825 TI - Role of sulfate reduction versus methanogenesis in terminal carbon flow in polluted intertidal sediment of waimea inlet, nelson, new zealand. AB - An investigation of the terminal anaerobic processes occurring in polluted intertidal sediments indicated that terminal carbon flow was mainly mediated by sulfate-reducing organisms in sediments with high sulfate concentrations (>10 mM in the interstitial water) exposed to low loadings of nutrient (equivalent to <10 kg of N . day) and biochemical oxygen demand (<0.7 x 10 kg . day) in effluents from different pollution sources. However, in sediments exposed to high loadings of nutrient (>10 kg of N . day) and biochemical oxygen demand (>0.7 x 10 kg . day), methanogenesis was the major process in the mediation of terminal carbon flow, and sulfate concentrations were low (/=0.96 for sediment with high sulfate, but in sediments with sulfate as little as 10 muM in the interstitial water, respiratory index values of 7 N,N-dimethylglycine + 1.5 acetate + 1.5 butyrate. E. limosum also grew on methanol and choline. The former was converted to acetate and butyrate, and the latter was converted to N,N dimethylethanolamine, acetate, and butyrate. The conditions for the quantitative determination of N,N-dimethylglycine by capillary tube isotachophoresis have been determined. PMID- 16345843 TI - Production of Naphthoquinone Mycotoxins and Taxonomy of Penicillium viridicatum. AB - Groups I and II of Penicillium viridicatum were further differentiated on the basis of synthesis of two mycotoxins, xanthomegnin and viomellein. Strains previously classified as group II produced these pigments, whereas those in group I did not. These napthoquinone pigments were quantitated by thin-layer chromatography and high-pressure liquid chromatography. A new mobile phase of toluene and acetic acid effected a baseline separation of the two components. It is proposed that such biochemical distinctions be incorporated into an artificial taxonomic scheme of use to nontaxonomists. PMID- 16345844 TI - Microbial Transformation of Quercetin by Bacillus cereus. AB - Biotransformation of quercetin was examined with a number of bacterial cultures. In the presence of a bacterial culture (Bacillus cereus), quercetin was transformed into two crystalline products, identified as protocatechuic acid and quercetin-3-glucoside (isoquercitrin). PMID- 16345845 TI - Enzymatic Basis for Overproduction of Tryptophan and Its Metabolites in Hansenula polymorpha Mutants. AB - 3-Deoxy-d-arabinoheptulosonate 7-phosphate (DAHP) synthetase and anthranilate synthetase are key regulatory enzymes in the aromatic amino acid biosynthetic pathway. The DAHP synthetase activity of Hansenula polymorpha was subject to additive feedback inhibition by phenylalanine and tyrosine but not by tryptophan. The synthesis of DAHP synthetase in this yeast was not repressed by exogenous aromatic amino acids, singly or in combinations. The activity of anthranilate synthetase was sensitive to feedback inhibition by tryptophan, but exogenous tryptophan did not repress the synthesis of this enzyme. Nevertheless, internal repression of anthranilate synthetase probably exists, since the content of this enzyme in H. polymorpha strain 3-136 was double that in the wild-type and less sensitive 5-fluorotryptophan-resistant strains. The biochemical mechanism for the overproduction of indoles by the 5-fluorotryptophan-resistant mutants was due primarily to a partial desensitization of the anthranilate synthetase of these strains to feedback inhibition by tryptophan. These results support the concept that inhibition of enzyme activities rather than enzyme repression is more important in the regulation of aromatic amino acid biosynthesis in H. polymorpha. PMID- 16345846 TI - Effects of nickel, cobalt, and molybdenum on performance of methanogenic fixed film reactors. AB - The conversion of acetic acid to methane and carbon dioxide by a mixed methanogenic population from an anaerobic fixed-film digestor was stimulated by the addition of nickel (100 nM) and cobalt (50 nM) and especially by the addition of these elements in combination. Molybdenum addition (50 nM) was only slightly stimulatory when added in combination with both nickel and cobalt. The addition of these trace metals to anaerobic fixed-film digestors, which treat food processing waste, greatly enhanced reactor performance. Total gas and methane productions were increased 42%, greater volumes of waste could be effectively treated, and reactor residence time was shortened. However, the lag period for reactor start-up was not reduced. Tests showed that reactor performance was increased because trace nutrient addition allowed accumulation of a thicker methanogenic fixed film. PMID- 16345847 TI - Brevianamides A and B Are Formed Only After Conidiation Has Begun in Solid Cultures of Penicillium brevicompactum. AB - In cultures of Penicillium brevicompactum grown on Czapek-Dox agar or on circles of dialysis membrane placed on Czapek-Dox agar, brevianamides A and B first appeared after conidiation had begun. The presence of these brevianamides imparted a yellow-green fluorescence to the penicilli and to the upper section of the conidiophores of actively conidiating cultures. Experiments with replacement cultures indicated that medium nutrient levels had little direct effect on brevianamide production. If P. brevicompactum was grown between two circles of dialysis membrane, no aerial hyphae were formed, and brevianamide production was not observed. Three brevianamide-minus mutants of P. brevicompactum were isolated. PMID- 16345848 TI - Microbiology of wetwood: importance of pectin degradation and clostridium species in living trees. AB - Wetwood samples from standing trees of eastern cottonwood (Populus deltoides), black poplar (Populus nigra), and American elm (Ulmus americana) contained high numbers of aerobic and anaerobic pectin-degrading bacteria (10 to 10 cells per g of wood). High activity of polygalacturonate lyase ( n-butanol > n-propanol > ethanol > glucose. Acetate and lactate caused no stimulation. PMID- 16345899 TI - Enhancement of the microbial dehalogenation of a model chlorinated compound. AB - A number of chlorinated aromatic and aliphatic compounds were dehalogenated when incubated with sewage. Preincubating the sewage with nonchlorinated organic substrates enhanced the subsequent dehalogenation of the chlorinated chemicals. Dehalogenation of 4-chloro-3,5-dinitrobenzoic acid (CDBA) in lake water occurred as a result of microbial growth both in the light in the absence of added nutrients and in the dark in the presence of acetate. No organism able to use CDBA as a carbon source was isolated. Axenic bacterial cultures and a nonaxenic Chlamydomonas culture released chloride from CDBA. The metabolism of CDBA by the latter culture, a process that was inhibited by 3-(3,4-dichlorophenyl)-1,1 dimethylurea, yielded a product that was identified as alpha-hydroxymuconic semialdehyde. This product of an apparent cometabolic transformation was mineralized by a strain of Streptomyces, thus suggesting that certain cometabolic products may not accumulate because they are carbon sources for other species. PMID- 16345900 TI - Kinetic explanation for accumulation of nitrite, nitric oxide, and nitrous oxide during bacterial denitrification. AB - The kinetics of denitrification and the causes of nitrite and nitrous oxide accumulation were examined in resting cell suspensions of three denitrifiers. An Alcaligenes species and a Pseudomonas fluorescens isolate characteristically accumulated nitrite when reducing nitrate; a Flavobacterium isolate did not. Nitrate did not inhibit nitrite reduction in cultures grown with tungstate to prevent formation of an active nitrate reductase; rather, accumulation of nitrite seemed to depend on the relative rates of nitrate and nitrite reduction. Each isolate rapidly reduced nitrous oxide even when nitrate or nitrite had been included in the incubation mixture. Nitrate also did not inhibit nitrous oxide reduction in Alcaligenes odorans, an organism incapable of nitrate reduction. Thus, added nitrate or nitrite does not always cause nitrous oxide accumulation, as has often been reported for denitrifying soils. All strains produced small amounts of nitric oxide during denitrification in a pattern suggesting that nitric oxide was also under kinetic control similar to that of nitrite and nitrous oxide. Apparent K(m) values for nitrate and nitrite reduction were 15 muM or less for each isolate. The K(m) value for nitrous oxide reduction by Flavobacterium sp. was 0.5 muM. Numerical solutions to a mathematical model of denitrification based on Michaelis-Menten kinetics showed that differences in reduction rates of the nitrogenous compounds were sufficient to account for the observed patterns of nitrite, nitric oxide, and nitrous oxide accumulation. Addition of oxygen inhibited gas production from NO(3) by Alcaligenes sp. and P. fluorescens, but it did not reduce gas production by Flavobacterium sp. However, all three isolates produced higher ratios of nitrous oxide to dinitrogen as the oxygen tension increased. Inclusion of oxygen in the model as a nonspecific inhibitor of each step in denitrification resulted in decreased gas production but increased ratios of nitrous oxide to dinitrogen, as observed experimentally. The simplicity of this kinetic model of denitrification and its ability to unify disparate observations should make the model a useful guide in research on the physiology of denitrifier response to environmental effectors. PMID- 16345901 TI - Specific Adhesion of Bacteria to Heterocysts of Anabaena spp. and Its Ecological Significance. AB - Two bacterial isolates, Pseudomonas sp. SL10 and Zoogloea sp. SL20, attach to heterocysts of Anabaena spp. with a high degree of selectivity, and this attachment can be expressed quantitatively in terms of adsorption isotherms. Adhesion of Pseudomonas sp. SL10 was restricted to a monolayer and exhibited a type I (Langmuir) isotherm, whereas adhesion of Zoogloea sp. SL20 involved multilayer attachment and exhibited a type II isotherm. The degree of adhesion by the bacteria to heterocysts of different Anabaena species may reflect the distribution and abundance of binding sites on the surface of different heterocysts. Both Pseudomonas sp. SL10 and Zoogloea sp SL20 promoted higher rates of acetylene reduction by Anabaena spp. under oxygenated culture conditions when compared with a cyanobacterial control. At ambient oxygen levels, however, only Zoogloea sp. SL20 stimulated acetylene reduction by Anabaena spp. PMID- 16345902 TI - Cellulose fermentation by a rumen anaerobic fungus in both the absence and the presence of rumen methanogens. AB - The fermentation of cellulose by an ovine rumen anaerobic fungus in the absence and presence of rumen methanogens is described. In the monoculture, moles of product as a percentage of the moles of hexose fermented were: acetate, 72.7; carbon dioxide, 37.6; formate, 83.1; ethanol, 37.4; lactate, 67.0; and hydrogen, 35.3. In the coculture, acetate was the major product (134.7%), and carbon dioxide increased (88.7%). Lactate and ethanol production decreased to 2.9 and 19%, respectively, little formate was detected (1%), and hydrogen did not accumulate. Substantial amounts of methane were produced in the coculture (58.7%). Studies with [2-C]acetate indicated that acetate was not a precursor of methane. The demonstration of cellulose fermentation by a fungus extends the range of known rumen organisms capable of participating in cellulose digestion and provides further support for a role of anaerobic fungi in rumen fiber digestion. The effect of the methanogens on the pattern of fermentation is interpreted as a shift in flow of electrons away from electron sink products to methane via hydrogen. The study provides a new example of intermicrobial hydrogen transfer and the first demonstration of hydrogen formation by a fungus. PMID- 16345903 TI - Entrance and growth of lactic Acid bacteria in gas-exchanged, brined cucumbers. AB - Entrance of lactic acid bacteria into the interior of brined cucumbers was found to be greatly influenced by gas composition of the cucumbers before brining. Exchange of the internal gas of fresh cucumbers with O(2) resulted in absorption of bacteria into the subsequently brined fruit within a few hours. Bacteria were absorbed into nonexchanged cucumbers to a lesser extent. Little bacterial absorption occurred in N(2)-exchanged cucumbers. Stomata of the cucumber skin appeared to be a likely port for bacterial entry. When Pediococcus cerevisiae or Lactobacillus plantarum cells were added to the brine of O(2)-exchanged cucumbers, the respective cell types colonized in large numbers within intercellular spaces and vascular elements of mesocarp tissue during fermentation of the cucumbers. Implications of these observations, particularly with regard to bloater formation in brined cucumbers, are discussed. PMID- 16345904 TI - Isolation, enumeration, and maintenance of rumen anaerobic fungi in roll tubes. AB - Anaerobic phycomycetous fungi have been isolated from rumen fluid by using the Hungate roll tube technique. Cultures on solid substrates in roll tubes can be stored for long periods, and fungal zoospores can be enumerated in roll tubes. PMID- 16345905 TI - Production of alcohol from apple pomace. AB - Production of ethyl alcohol from apple pomace with a Montrachet strain of Saccharomyces cerevisiae is described. More than 43 g of the ethyl alcohol could be produced per kg of apple pomace fermented at 30 degrees C in 24 h. The fermentation efficiency of this process was approximately 89%. PMID- 16345906 TI - Autolytic Activity and an Autolysis-Deficient Mutant of Clostridium acetobutylicum. AB - The optimum conditions for autolysis and autoplast formation in Clostridium acetobutylicum P262 have been defined. Autolysis was optimal at pH 6.3 in 0.04 M sodium phosphate buffer, and the bacterium produced latent and active forms of an autolytic enzyme. The ability of cells to autolyze decreased sharply when cultures entered the stationary phase. Autoplasts were induced by 0.25 to 0.5 M sucrose and were stable in media containing sucrose, CaCl(2), and MgCl(2). A pleiotropic autolysis-deficient mutant (lyt-1) was isolated. The mutant produced less autolysin than did the parent P262 strain, and it had an altered cell wall which was more resistant to both its own and P262 autolysins. The mutant formed long chains of cells, and lysozyme was required for the production of autoplasts. Growth of the P262 strain or the lyt-1 mutant was inhibited by the same concentrations of penicillin, ampicillin, and vancomycin. The lyt-1 mutant strain treated with the minimum growth-inhibitory concentration of penicillin autolyzed upon the addition of wild-type autolysin to the autolysis buffer at the same rate as did the untreated P262 strain. Chloramphenicol did not protect the penicillin treated lyt-1 cells against autolysis enhanced by exogenous wild-type autolysin. PMID- 16345907 TI - Use of a fractional factorial design to evaluate interactions of environmental factors affecting biodegradation rates. AB - For investigation of main and interactive effects of six experimentally controlled environmental factors on phenol biodegradation in a shake-flask system, a largely neglected statistical procedure was applied. A major benefit resulting from the application of the orthogonal, fractional factorial design is that the number of experiments necessary to evaluate multifactor interactions is limited. In our investigation, the required number of experiments was reduced to 81 from the 324 necessary with conventional factorial designs; information was sacrificed for only 3 of 15 possible two-factor interactions. Six experimentally controlled factors were investigated at two or three treatment levels each; the six factors were (1) amount of phenol substrate, (2) amount of bacterial inoculum, (3) filtration of inoculum, (4) type of basal salts medium, (5) initial pH of basal salts medium, and (6) flask closure. Significant main effects were found for factors 1, 2, and 4; whereas significant interactive effects were found only for factor 2 with factor 3 and for factor 2 with factor 5. Our results suggest that the application of these statistical designs will greatly reduce the number of experiments necessary to evaluate multifactor effects on degradation rates during optimization of both hazard screening systems and waste treatment systems. PMID- 16345908 TI - Evidence for Plasmid Linkage of Restriction and Modification in Streptococcus cremoris KH. AB - Restriction and modification have been demonstrated in Streptococcus cremoris KH cells when infected by Streptococcus lactis C2 phage (designated c2) at an efficiency of plating of 2 x 10. The growth of c2 phage through KH cells produces modified progeny phage capable of unrestricted growth on KH cells. The ability of single-colony isolates of S. cremoris KH cultures to restrict and modify c2 phage was found to be variable. From 2 to 6.5% of colonies isolated were partially deficient in restrictive capacity, permitting a greater plaquing ability by c2 phage of 1.8 to 2.9 log cycles. No completely restrictionless mutants were isolated from 1,000 colonies examined. Mutants were shown to be deficient in both restriction and modification capabilities of the same specificity. The frequent occurrence of a genotypic change that resulted in the loss of both restriction and modification capacities indicated the involvement of plasmid deoxyribonucleic acid in genetically determining this specific restriction and modification system. S. cremoris KH was found to harbor 11 plasmid molecules, with molecular weights (x10) estimated to be 50, 41, 24, 18, 10, 7.4, 3.3, 3.0, 2.8, 2.5, and 1.5. Of the 27 mutants examined, 25 were missing the 10-megadalton plasmid. This consistent plasmid difference among the majority of mutants isolated supports the involvement of this plasmid in restriction and modification. Plasmid linkage of restriction and modification systems provides a genetic mechanism for the rapid development of phage-sensitive starter cultures due to the inherent instability of extrachromosomal elements. PMID- 16345909 TI - Survival of Rhizobium in Acid soils. AB - A Rhizobium strain nodulating cowpeas did not decline in abundance after it was added to sterile soils at pH 6.9 and 4.4, and the numbers fell slowly in nonsterile soils at pH 5.5 and 4.1. A strain of R. phaseoli grew when added to sterile soils at pH 6.7 and 6.9; it maintained large, stable populations in soils of pH 4.4, 5.5, and 6.0, but the numbers fell markedly and then reached a stable population size in sterile soils at pH 4.3 and 4.4. The abundance of R. phaseoli added to nonsterile soils with pH values of 4.3 to 6.7 decreased similarly with time regardless of soil acidity, and the final numbers were less than in the comparable sterile soils. The minimum pH values for the growth of strains of R. meliloti in liquid media ranged from 5.3 to 5.9. Two R. meliloti strains, which differed in acid tolerance for growth in culture, did not differ in numbers or decline when added to sterile soils at pH 4.8, 5.2, and 6.3. The population size of these two strains was reduced after they were introduced into nonsterile soils at pH 4.8, 5.4, and 6.4, and the number of survivors was related to the soil pH. The R. meliloti strain that was more acid sensitive in culture declined more readily in sterile soil at pH 4.6 than did the less sensitive strain, and only the former strain was eliminated from nonsterile soil at pH 4.8; however, the less sensitive strain also survived better in limed soil. The cell density of the two R. meliloti strains was increased in pH 6.4 soil in the presence of growing alfalfa. The decline and elimination of the tolerant, but not the sensitive, strain was delayed in soil at pH 4.6 by roots of growing alfalfa. PMID- 16345910 TI - Evidence for coexistence of two distinct functional groups of sulfate-reducing bacteria in salt marsh sediment. AB - Oxidation of acetate in salt marsh sediment was inhibited by the addition of fluoroacetate, and also by the addition of molybdate, an inhibitor of sulfate reducing bacteria. Molybdate had no effect upon the metabolism of acetate in a freshwater sediment in the absence of sulfate. The inhibitory effect of molybdate on acetate turnover in the marine sediment seemed to be because of its inhibiting sulfate-reducing bacteria which oxidized acetate to carbon dioxide. Sulfide was not recovered from sediment in the presence of molybdate added as an inhibitor of sulfate-reducing bacteria, but sulfide was recovered quantitatively even in the presence of molybdate by the addition of the strong reducing agent titanium chloride before acidification of the sediment. Reduction of sulfate to sulfide by the sulfate-reducing bacteria in the sediment was only partially inhibited by fluoroacetate, but completely inhibited by molybdate addition. This was interpreted as showing the presence of two functional groups of sulfate-reducing bacteria-one group oxidizing acetate, and another group probably oxidizing hydrogen. PMID- 16345911 TI - Facile method for monitoring inhibition of anaerobic spore outgrowth. AB - A device is presented for the laboratory monitoring of spore outgrowth under controlled temperature and anaerobic conditions. Alterations in pH, redox potential, headspace composition, and optical density are followed as the activated spores grow out into vegetative cells. An interlock system allows the addition of test solutions or the removal of medium under anaerobic conditions. The device may also be used for rapid (<4 h) chemical inhibition studies or adapted for temperature injury studies of aerobic or anaerobic cells. Data on outgrowth of Clostridium sporogenes and inhibition by nitrite solutions are presented. PMID- 16345913 TI - Isolation of a Strain of Clostridium thermoaceticum Capable of Growth and Acetic Acid Production at pH 4.5. AB - Using a series of pH controlled batch fermentations operated in a fed-batch mode and adaptation and selection techniques where pH and acetic acid provided the selective pressures, we isolated a culture of Clostridium thermoaceticum that can grow and produce acetic acid at pH 4.5. At pH 4.5 the fastest mass doubling time was 36 h, and the highest acetic acid concentration reached was 4.5 g/liter. Generally, as the pH was decreased from 6.0 and the initial acetic acid concentration increased, the mass doubling time increased, and the final acetic acid concentration decreased. These observations can be explained in terms of inhibition by the free acetic acid concentration at a given pH, relative to the total acetic acid concentration (free acid plus acetate ion). We have thus reached one of the criteria determined by us to be required for an economically viable fermentation acetic acid process, i.e., pH 4.5. A second requirement for a mass doubling time of about 7 h (0.1/h dilution rate) can probably be reached by selection in continuous culture. The final requirement for an acetic acid concentration of 50 g/liter will be the most difficult to achieve in view of the organism's sensitivity to low concentrations of free acetic acid. PMID- 16345912 TI - Effect of temperature on heterotrophic glucose uptake, mineralization, and turnover rates in lake sediments. AB - The V(max) and turnover rates (TR) of [U-C]glucose uptake and mineralization of Lake Kinneret (Israel) sediment are temperature dependent. The following activation energies were determined: glucose uptake, approximately 15,000 cal (62,760 J); TR of glucose uptake, approximately 10,000 cal (41,840 J); glucose mineralization, 7,500 to 15,000 cal (31,380 to 62,760 J); and TR of glucose mineralization, approximately 15,000 cal. Q(10) values varied as follows: glucose uptake, approximately 2.3; TR of glucose uptake, approximately 1.8; and glucose mineralization, approximately 2.5. K + S(n) values increased slightly with temperature and might reflect an increased K with increased temperatures. Glucose respiration/uptake ratios were low (9.5 to 12%) and were apparently not greatly influenced by the presence or absence of oxygen or by different assay temperatures. Aerobic or anaerobic sediments assayed under either aerobic or anaerobic conditions did not exhibit greatly different V(max), TR, or K + S(n) values. PMID- 16345914 TI - Population of aerobic heterotrophic nitrogen-fixing bacteria associated with wetland and dryland rice. AB - Nitrogen-fixing activity and populations of nitrogen-fixing bacteria associated with two varieties of rice grown in dryland and wetland conditions were measured at various growth stages during the dry season. Acetylene reduction activities were measured both in the field and for the hydroponically grown rice, which was transferred from the field to water culture 1 day before assay. The activities measured by both methods were higher in wetland than in dryland rice. The population of nitrogen-fixing heterotrophic bacteria associated with rhizosphere soil, root, and basal shoots was determined by the most probable number method with semisolid glucose-yeast extract and semisolid malate-yeast extract media. The number of nitrogen-fixing bacteria was higher in wetland conditions than in dryland conditions. The difference between two conditions was most pronounced in the population associated with the basal shoot. The glucose medium gave higher counts than did the malate medium. Colonies were picked from tryptic soy agar plates, and their nitrogen-fixing activity was tested on a semisolid glucose yeast extract medium. The incidence of nitrogen-fixing bacteria among aerobic heterotrophic bacteria in association with rhizosphere soil, root, and basal shoots was much lower in dryland rice than in wetland rice. PMID- 16345915 TI - Effects of petroleum hydrocarbons on plant litter microbiota in an arctic lake. AB - The effects of petroleum hydrocarbons on the microbial community associated with decomposing Carex leaf litter colonized in Toolik Lake, Alaska, were examined. Microbial metabolic activity, measured as the rate of acetate incorporation into lipid, did not vary significantly from controls over a 12-h period after exposure of colonized Carex litter to 3.0 ml of Prudhoe Bay crude oil, diesel fuel, or toluene per liter. ATP levels of the microbiota became elevated within 2 h after the exposure of the litter to diesel fuel or toluene, but returned to control levels within 4 to 8 h. ATP levels of samples exposed to Prudhoe Bay crude oil did not vary from control levels. Mineralization of specifically labeled C [lignin]-lignocellulose and C-[cellulose]-lignocellulose by Toolik Lake sediments, after the addition of 2% (vol/vol) Prudhoe Bay crude oil, motor oil, diesel fuel, gasoline, n-hexane, or toluene, was examined after 21 days of incubation at 10 degrees C. Diesel fuel, motor oil, gasoline, and toluene inhibited C-[lignin]-lignocellulose mineralization by 58, 67, 67, and 86%, respectively. Hexane-treated samples displayed an increase in the rate of C [lignin]-lignocellulose mineralization of 33%. C-[cellulose]-lignocellulose mineralization was inhibited by the addition of motor oil or toluene by 27 and 64%, respectively, whereas diesel fuel-treated samples showed a 17% increase in mineralization rate. Mineralization of the labeled lignin component of lignocellulose appeared to be more sensitive to hydrocarbon perturbations than was the labeled cellulose component. PMID- 16345916 TI - Characterization of bacteria from a Swine manure digester. AB - One-hundred thirty bacteria isolated from a swine manure digester were predominately gram-positive anaerobes which were tentatively classified into the following genera: Peptostreptococcus, Eubacterium, Bacteroides, Lactobacillus, Peptococcus, Clostridium, and Streptococcus plus two unidentified groups. The major fermentation products formed by these organisms included acetate, propionate, succinate, lactate, and ethanol, singly or in various combinations. Acetate was the sole end product of several groups. Few of the isolates (14%) reduced the pH below 6.0. The predominate bacteria appear to differ from the predominate organisms isolated from other anaerobic ecosystems. PMID- 16345917 TI - Biodegradation of glycidol and glycidyl nitrate. AB - When calcium hydroxide is used to desensitize glycerol trinitrate (nitroglycerine)-containing waste streams, the epoxides glycidol and glycidyl nitrate are formed. The epoxide rings of both compounds are unstable to heat in aqueous solutions, and they open to form glycerol 1-mononitrate and presumably glycerol. These transformations were accelerated by microbial activity. Glycerol 1-mononitrate was slowly denitrated to form glycerol. Glycidol and glycidyl nitrate caused base-pair substitutions in the Ames test for mutagenicity, whereas glycerol 1-mononitrate tests were negative. PMID- 16345918 TI - Assimilatory Sulfur Metabolism in Marine Microorganisms: Sulfur Metabolism, Protein Synthesis, and Growth of Alteromonas luteo-violaceus and Pseudomonas halodurans During Perturbed Batch Growth. AB - The antibiotic protein synthesis inhibitor chloramphenicol specifically blocked the incorporation of [S]sulfate into the residue protein of two marine bacteria, Pseudomonas halodurans and Alteromonas luteo-violaceus. Simultaneous inhibition of total protein synthesis occurred, but incorporation of S into low-molecular weight organic compounds continued. A. luteo-violaceus rapidly autolyzed, with similar reduction in cell counts, total culture protein and cellular sulfur, whereas P. halodurans remained viable. Treatment with chloramphenicol, growth during nitrogen and carbon limitation, and the carbon and energy sources used for growth did not alter the sulfur content of P. halodurans protein. The mean value (1.09%, by weight), representing a wide variety of environmentally relevant growth conditions, was in agreement with model protein composition. The variability of cellular composition of P. halodurans and A. luteo-violaceus is discussed with respect to the measurement of bacterial growth in natural environments. Total carbon and nitrogen per cell varied greatly (coefficient of variation, ca. 100%) depending on growth conditions. Variation in total sulfur and protein per cell was much less (coefficient of variation, <50%), but the least variation was found for sulfate incorporation into residue protein (coefficient of variation, ca. 15%). Thus, sulfate incorporation into residue protein can be used as an accurate measurement of de novo protein synthesis in these bacteria. PMID- 16345919 TI - Assimilatory sulfur metabolism in marine microorganisms: considerations for the application of sulfate incorporation into protein as a measurement of natural population protein synthesis. AB - The sulfur content of residue protein was determined for pure cultures of Nitrosococcus oceanus, Desulfovibrio salexigens, 4 mixed populations of fermentative bacteria, 22 samples from mixed natural population enrichments, and 11 nutritionally and morphologically distinct isolates from enrichments of Sargasso Sea water. The average 1.09 +/- 0.14% (by weight) S in protein for 13 pure cultures agrees with the 1.1% calculated from average protein composition. An operational value encompassing all mixed population and pure culture measurements has a coefficient of variation of only 15.1% (n = 41). Short-term [S]sulfate incorporation kinetics by Pseudomonas halodurans and Alteromonas luteoviolaceus demonstrated a rapid appearance of S in the residue protein fraction which was well modelled by a simple exponential uptake equation. This indicates that little error in protein synthesis determination results from isotope dilution by endogenous pools of sulfur-containing compounds. Methionine effectively competed with sulfate for protein synthesis in P. halodurans at high concentrations (10 muM), but had much less influence at 1 muM. Cystine competed less effectively with sulfate, and glutathione did not detectably reduce sulfate S incorporation into protein. [S]sulfate incorporation was compared with [C]glucose assimilation in a eutrophic brackish-water environment. Both tracers yielded similar results for the first 8 h of incubation, but a secondary growth phase was observed only with S. Redistribution of C from low-molecular-weight materials into residue protein indicated additional protein synthesis. [S]sulfate incorporation into residue protein by marine bacteria can be used to quantitatively measure bacterial protein synthesis in unenriched mixed populations of marine bacteria. PMID- 16345920 TI - Diversity dynamics of marine bacteria studied by immunofluorescent staining on membrane filters. AB - Of 34 strains of marine bacteria isolated on a general seawater medium, 5 were selected for detailed studies of their population dynamics in the plankton. The isolates were characterized as Aeromonas sp., Chromobacterium cf. lividum, Vibrio sp., and two Pseudomonas spp. Specific antibodies were produced by immunization of rabbits, and bacterial cells were stained on black Uni-Pore membrane filters by an indirect immunofluorescent staining procedure. The method proved to be very specific and practical for use in a large-scale field sampling program. Growth of all five isolates was stimulated by high values for net primary production, chlorophyll a, and dissolved organic carbon. Calculation of a diversity index based on specific and total counts is proposed as a way of characterizing the dynamics of organotrophic bacterial populations in the sea. PMID- 16345921 TI - Contribution of particle-bound bacteria to total microheterotrophic activity in five ponds and two marshes. AB - We examined the abundance and heterotrophic uptake of bacteria attached to particulate matter suspended in five coastal ponds and two marshes near Woods Hole, Mass. Although the number of particle-bound bacteria was low (<10%), these bacteria incorporated a large proportion (>40%) of [C]glucose and [C]glutamate in selected aquatic systems. The uptake per cell was significantly higher for epibacteria than for unattached bacteria in all systems. Two groups of the aquatic environments sampled were statistically different in the contribution made by particle-bound bacteria to total bacterial abundance and to total assimilation of [C]glucose and [C]glutamate. Particle-bound bacteria were more important in those waters with a high particle concentration and not flushed regularly by tides than in waters with a low particle concentration and flushed regularly. PMID- 16345922 TI - Feasibility of Fe Autoradiography as Performed on N(2)-Fixing Anabaena spp. Populations and Associated Bacteria. AB - Fe emits low-energy X rays and Auger electrons by electron capture decay. Auger electrons are useful for autoradiographic examination of Fe incorporation among microbial communities. Attainable resolution, in terms of silver grain deposition, is excellent and comparable to H. Two known Fe-demanding processes, photosynthetic CO(2) fixation and N(2) fixation, were examined by autoradiography of Anabaena populations. During photosynthetically active (illuminated) N(2) fixing periods, biological incorporation of FeCl(3) by vegetative cells and heterocysts was evident. When N(2) fixation was suppressed by NH(4) additions, heterocysts revealed no incorporation of Fe. Conversely, when N(2)-fixing Anabaena filaments were placed in darkness, Fe incorporation decreased in vegetative cells, whereas heterocysts showed sustained rates of Fe incorporation. Bacteria actively incorporated Fe under both light and dark conditions. The chelated (by Na(2)-ethylenediaminetetraacetate) form of FeCl(3) was more readily incorporated than the nonchelated form. Furthermore, abiotic adsorption of Fe to filters and nonliving particles proved lower when chelated Fe was used in experiments. Fe autoradiography is useful for observing the fate and cellular distribution of various forms of Fe among aquatic microbial communities. PMID- 16345923 TI - Localized Tetrazolium Reduction in Relation to N(2) Fixation, CO(2) Fixation, and H(2) Uptake in Aquatic Filamentous Cyanobacteria. AB - The aquatic filamentous cyanobacteria Anabaena oscillarioides and Trichodesmium sp. reveal specific cellular regions of tetrazolium salt reduction. The effects of localized reduction of five tetrazolium salts on N(2) fixation (acetylene reduction), CO(2) fixation, and H(2) utilization were examined. During short-term (within 30 min) exposures in A. oscillarioides, salt reduction in heterocysts occurred simultaneously with inhibition of acetylene reduction. Conversely, when salts failed to either penetrate or be reduced in heterocysts, no inhibition of acetylene reduction occurred. When salts were rapidly reduced in vegetative cells, CO(2) fixation and H(2) utilization rates decreased, whereas salts exclusively reduced in heterocysts were not linked to blockage of these processes. In the nonheterocystous genus Trichodesmium, the deposition of reduced 2,3,5-triphenyl-2-tetrazolium chloride (TTC) in the internal cores of trichomes occurs simultaneously with a lowering of acetylene reduction rates. Since TTC deposition in heterocysts of A. oscillarioides occurs contemporaneously with inhibition of acetylene reduction, we conclude that the cellular reduction of this salt is of use in locating potential N(2)-fixing sites in cyanobacteria. The possible applications and problems associated with interpreting localized reduction of tetrazolium salts in cyanobacteria are presented. PMID- 16345924 TI - Transposon mutagenesis in rhizobia which can nodulate both legumes and the nonlegume parasponia. AB - The successful use of transposon mutagenesis in a slow-growing Rhizobium permitted the isolation of auxotrophic and symbiotically defective mutants. In these mutants, it was shown that different single-copy Tn5 insertions had occurred. Mutant phenotypes indicated that the control of nitrogenase activity in vitro and in planta is different. PMID- 16345925 TI - Influence of pH on Inhibitory Activity of Acetic Acid on Osmophilic Yeasts Used in Brine Fermentation of Soy Sauce. AB - The inhibitory activity of acetic acid on osmophilic yeasts (Saccharomyces rouxii and Torulopsis versatilis) used for the brine fermentation of soy sauce increased remarkably as the pH of the medium was lowered. This increased toxicity could not be attributed to a hydrogen ion effect alone. PMID- 16345926 TI - Medium for enhanced growth of bacteria from a Swine manure digester. AB - Most bacterial strains isolated from a swine manure digester grew sufficiently to permit transfer of cultures, but not characterization. Substrates, crude extracts, growth factors, and electron acceptors were evaluated for growth promotion. The growth of all but one group of the isolates was substantially increased with a medium containing glucose, cellobiose, soluble starch, pyruvate, peptone, yeast extract, minerals, volatile acids, vitamins, hemin plus vitamins K(1) and K(3), sodium bicarbonate, cysteine, and digester fluid. The strains require both known and unknown factors (in crude extracts) for maximal growth. PMID- 16345927 TI - Partial purification and characterization of hemolysin from a psychrotrophic kanagawa-positive marine Vibrio. AB - Psychrotrophic Kanagawa-positive marine vibrios were isolated from soft-shelled clams (Mya arenaria) collected in Yaquina Bay, Oreg. The 235 vibrio isolates obtained were screened for Gram reaction and morphology, Kanagawa reaction on Wagastsuma agar, and response to selected biochemical tests. The vibrio selected for further study was grown in broth, and the hemolysin was precipitated from a cleared supernatant with solid ammonium sulfate. The hemolytic substance was partially purified by DEAE-cellulose and Sephadex G-100 column chromatography. The hemolysin contained protein essential for activity, was thermolabile, and was more active against rabbit erythrocytes at 37 degrees C than at lower temperatures. The molecular weight was estimated at 55,000 by using a Sephadex G 100 column. Hemolytic activity was partially inactivated by gangliosides and lowered against horse erythrocytes. The hemolysin did not react with antibody prepared against vibriolysin from Vibrio parahaemolyticus WP-1 by the Ouchterlony method. The hemolysin was high in aspartic and glutamic acids and low in arginine and histidine. Electrophoresis on a sodium dodecyl sulfate-polyacrylamide gel gave three major bands. The hemolysin from a psychrotrophic vibrio and the hemolytic exotoxin of V. parahaemolyticus had some similar and dissimilar characteristics. The possibility that a Vibrio sp. other than V. parahaemolyticus might serve as the reservoir for the Kanagawa phenotype is discussed. PMID- 16345928 TI - Terminal reactions in the anaerobic digestion of animal waste. AB - An anaerobic mesophilic digestor was operated using beef cattle waste (diluted to 5.75% volatile solids) as substrate; retention time was 10 days with daily batch feed. Volatile solids destruction was 36%. Daily gas production rate was 1.8 liters of gas (standard temperature and pressure) per liter of digestor contents (0.99 liters of CH(4) per liter of digestor contents). Acetate turnover was measured, and it was calculated that 68% of the CH(4) was derived from the methyl group of acetate. When the methanogenic substrates acetic acid or H(2)/CO(2) were added to the digestor on a continuous basis, the microflora were able to adapt and convert them to terminal products while continuing to degrade animal waste to the same extent as without additions. The methanogenic substrates were added at a rate at least 1.5 times the microbial production rate which was measured in the absence of added substrates. Added acetate was converted directly to CH(4) by acetoclastic methanogens; H(2) addition greatly stimulated acetate production in the digestor. A method is described for the measurement of acetate turnover in batch-fed digestors. PMID- 16345929 TI - Method for studying microbial biofilms in flowing-water systems. AB - A method for the study of microbial biofilms in flowing-water systems was developed with special reference to the flow conditions in electrochemical concentration cells. Seawater was circulated in a semiclosed flow system through biofilm reactors (3 cm s) with microscope cover slips arranged in lamellar piles parallel with the flow. At fixed time intervals cover slips with their biofilm were removed from the pile, stained with crystal violet, and mounted on microscope slides. The absorbances of the slides were measured at 590 nm and plotted against time to give microbial biofilm development. From calibration experiments a staining time of 1 min and a rinse time of 10 min in a tap water flow (3 cm s) were considered sufficient. When an analysis of variance was performed on biofilm development data, 78% of the total variance was found to be due to random natural effects; the rest could be explained by experimental effects. The absorbance values correlated well with protein N, dry weight, and organic weight in two biofilm experiments, one with a biofilm with a high (75%) and one with a low ( approximately 25%, normal) inorganic content. Comparisons of regression lines revealed that the absorbance of the stained biofilms was an estimate closely related to biofilm dry weight. PMID- 16345930 TI - Two Naturally Occurring Nuclear Polyhedrosis Virus Variants of Neodiprion sertifer Geoffr. (Hymenoptera; Diprionidae). AB - Two isolates of Neodiprion sertifer (Geoffr.) nuclear polyhedrosis virus from Britain and North America were compared using three biochemical techniques. Alkaline protease assays of polyhedra revealed the presence of endogenous enzyme activity in the British isolate but not in the North American isolate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of virus particle structural polypeptides revealed only minor differences, with the exception that the North American virus was contaminated with polyhedral protein. The restriction endonucleases SalI, HindIII, and HpaII were used as a definitive method of distinguishing the two variants, with all endonucleases achieving this to a greater or lesser extent. The possible significance of all of these observations is discussed in terms of their possible influence on the registration and field application of this virus. PMID- 16345931 TI - Growth of Eubacterium limosum with Carbon Monoxide as the Energy Source. AB - Eubacterium limosum grew with CO as the sole source of energy and formed acetate and CO(2) as the major products. The generation time on CO was 7 h. Uninhibited growth occurred in cultures containing 50% CO or less, but growth occurred at all concentrations tested (i.e., up to 75% CO). The pH optimum for growth was 7.0 to 7.2, whereas growth was poor at a pH below 6.7. CO(2) stimulated growth on CO. CO was preferentially utilized when both CO and H(2) were present. PMID- 16345932 TI - Production of Maltase by Wild-Type and a Constitutive Mutant of Saccharomyces italicus. AB - The production of maltase, an inducible and repressible catabolic enzyme in Saccharomyces italicus, was studied and compared in batch, fed-batch, and continuous fermentations. Tight genetic controls on maltase synthesis limited the effect of environmental manipulations such as fed-batch or continuous culture in enhancement of maltase synthesis, and neither approach was able to improve the performance above the batch process for maltase production. S. italicus was mutated, and a constitutive producer of maltase was isolated. The mutant was detected by its ability to grow on sucrose, which is a noninducing substrate that is hydrolyzed by maltase; S. italicus does not possess invertase and will not normally grow on sucrose. Maltase production by this mutant was studied during growth on sucrose in batch and continuous cultures and marked improvement in enzyme productivity was observed. The specific activity of maltase produced by this mutant was more than twice that of the parent wild type: 2,210 and 1,370 U/g of cells for the mutant versus 890 and 510 U/g of cells for the wild type in batch and continuous cultures, respectively. Maltase specific productivity was increased from 74 to 288 U/g of cells per h by switching from batch growth of the wild type to continuous cultivation of the mutant. PMID- 16345933 TI - Membrane Filter Method for Enumeration of Acinetobacter calcoaceticus from Environmental Waters. AB - A membrane filter method was developed and evaluated for the quantitative recovery of Acinetobacter calcoaceticus from environmental waters. The procedure utilized a mineral medium, with sodium acetate and potassium nitrate as the carbon and nitrogen sources, respectively. Formic acid was included to enhance the recovery of A. calcoaceticus and to inhibit background growth. The medium was incubated for 46 h at 30 degrees C, after which fermentation and cytochrome oxidase tests were performed on the colonies as they appeared on the membrane. Background microbial growth decreased on the average by 1.77 orders of magnitude. An essentially quantitative recovery relative to that on nutrient agar spread plates was obtained from freshly prepared suspensions of eight A. calcoaceticus strains in filter-sterilized pond water and from suspensions of five of the strains held for up to 96 h in filter-sterilized pond water at 15 and 22 degrees C. Markedly reduced relative recoveries were obtained with the three remaining strains. However, these three strains, in contrast to the first five, not only did not grow, but also decreased in number in the eutrophic, filter-sterilized pond water. The confirmation rate of presumptive A. calcoaceticus colonies was 95%, whereas 8% of the presumptively negative colonies were A. calcoaceticus. The precision of the method did not exceed that expected from random error alone. Densities of A. calcoaceticus in freshwaters ranged from <1 to 7.9 x 10 organisms per 100 ml and were about 10 organisms per 100 ml in raw sewage. PMID- 16345934 TI - Correlations between predominant heterotrophic bacteria and physicochemical water quality parameters in two canadian rivers. AB - The heterotrophic bacterial populations in two contrasting rivers have been examined over a period of 1 year. The populations were analyzed (i) as total heterotrophic counts, (ii) as species numbers, using numerical taxonomy, (iii) by diversity indices, and (iv) by factor analysis. Isolates were obtained by plating directly from water samples and by chemostat enrichment. Four factors emerged which profiled the bacterial community and were common to both rivers. They were, in order of decreasing importance, fermentative metabolism, inorganic nitrogen metabolism, fluorescence-oxidative metabolism, and lack of starch hydrolysis. Several factors produced significant correlations with a range of physicochemical parameters, which were also measured. The correlations suggested an intricate algal-bacterial interaction. The oxidative metabolism factor correlated with rainfall in one river, suggesting that the oxidative bacteria may be washed in from the surrounding land. In the other river, the oxidative-fermentative factor correlated negatively with sunshine. Factor analysis was the most effective method for revealing correlations between bacterial characteristics and the environmental parameters; however, the use of a variety of methods provided more insight into the ecological aspects. PMID- 16345935 TI - Isolation of Plasmid DNA from Butyrivibrio fibrisolvens. AB - A procedure based on successive precipitation of cell lysates with sodium dodecyl sulfate-NaCl and polyethylene glycol 6000 was developed which allows the isolation of plasmid DNA from Butyrivibrio fibrisolvens. A survey of B. fibrisolvens strains isolated from the bovine rumen showed that plasmids are a common feature of this species. PMID- 16345936 TI - Effects of Polyenes, Detergents, and Other Potential Membrane Perturbants on an Osmotolerant Yeast, Saccharomyces rouxii. AB - The osmotolerance of Saccharomyces rouxii 48-28 was confirmed with both NaCl- and KCl-fortified growth media, with more tolerance being exhibited for the potassium salt. Washed and buffered cells from unfortified medium were challenged with a variety of compounds (and also with physical treatments) that potentially would elicit membrane perturbations. The efficacy of these brief treatments was judged primarily by monitoring subsequent viability. Change in the degree of expression of beta-fructofuranosidase (EC 3.2.1.26), which is cryptic in young cells of S. rouxii, was a second criterion. There was a linear correlation between cell death and enzyme expression for treatments with polyenes, detergents, some organic solvents which did not denature the enzyme, and various freeze-thaw regimens in graded amounts of glycerol. The species is relatively insensitive to polyene antimycotics, the order of decreasing effect being filipin, nystatin, and amphotericin B. S. rouxii was found to be less sensitive to osmotic shock than is Saccharomyces cerevisiae, but in neither species is beta-fructofuranosidase released to the medium. The sensitivity of S. rouxii to ionic detergents, but not to nonionic detergents, was rationalized as being due to cell wall discrimination against larger micelles for the nonionic examples. This was confirmed by showing that protoplasts were sensitive to both classes. In cultures older than 5 days the normal agreement between colony-forming units and methylene blue exclusion (another test of viability) no longer held. Delayed fermentation of sucrose by S. rouxii, which is a diagnostic feature of the species, is explained by death of some cells, expression of their beta-fructofuranosidase, and utilization of the monosaccharides by the surviving cells. PMID- 16345937 TI - Reduction of ferric iron in anaerobic, marine sediment and interaction with reduction of nitrate and sulfate. AB - Studies were carried out to elucidate the nature and importance of Fe reduction in anaerobic slurries of marine surface sediment. A constant accumulation of Fe took place immediately after the endogenous NO(3) was depleted. Pasteurized controls showed no activity of Fe reduction. Additions of 0.2 mM NO(3) and NO(2) to the active slurries arrested the Fe reduction, and the process was resumed only after a depletion of the added compounds. Extended, initial aeration of the sediment did not affect the capacity for reduction of NO(3) and Fe, but the treatments with NO(3) increased the capacity for Fe reduction. Addition of 20 mM MoO(4) completely inhibited the SO(4) reduction, but did not affect the reduction of Fe. The process of Fe reduction was most likely associated with the activity of facultative anaerobic, NO(3)-reducing bacteria. In surface sediment, the bulk of the Fe reduction may be microbial, and the process may be important for mineralization in situ if the availability of NO(3) is low. PMID- 16345939 TI - Disposition of Mycophenolic Acid, Brevianamide A, Asperphenamate, and Ergosterol in Solid Cultures of Penicillium brevicompactum. AB - When Penicillium brevicompactum was grown between layers of dialysis membrane on Czapek-Dox agar for 3 days, no synthesis of mycophenolic acid, brevianamide A, asperphenamate, or ergosterol occurred. After removal of the uppermost membrane layer, aerial mycelium developed and all four metabolites were formed. The bulk of the mycophenolic acid that was formed was excreted into the medium, the bulk of the brevianamide A and asperphenamate was found in the aerial hyphae, and ergosterol was found in both vegetative and aerial mycelia. PMID- 16345938 TI - Determination of the Number of Respiring Thiobacillus ferrooxidans Cells in Water Samples by Using Combined Fluorescent Antibody-2-(p-Iodophenyl)-3-(p-Nitrophenyl) 5-Phenyltetrazolium Chloride Staining. AB - Fluorescent antibody staining was combined with 2-(p-iodophenyl)-3-(p nitrophenyl)-5-phenyltetrazolium chloride reduction in a procedure termed FAINT to allow for the direct microscopic determination of specific actively respiring populations of bacteria in a variety of aquatic habitats. The FAINT procedure is simple, precise, and appropriate for use in a wide variety of autecological studies. The distribution of Thiobacillus ferrooxidans was examined by FAINT enumerations in both acidic and nonacidic sites. Comparisons among the FAINT technique and fluorescent antibody staining alone or most-probable-number determinations in 9K broth showed that the use of most-probable-number determinations resulted in an underestimation of the number of viable cells by one to three orders of magnitude, whereas fluorescent antibody counts resulted in an overestimation of the number of viable T. ferrooxidans. The amount of difference was not consistent but varied, depending on the sample site. PMID- 16345940 TI - Lysogenic strains of lactic Acid streptococci and lytic spectra of their temperate bacteriophages. AB - A total of 113 strains of mesophilic strains lactic streptococci of the species Streptococcus lactis, S. lactis subsp. diacetilactis, and S. cremoris, chosen from 291 strains that had been previously classified into six groups on the basis of their sensitivity to 132 virulent phages, were subjected to induction with mitomycin C. Among these strains, 43% produced phages capable of forming plaques of lysis on an indicator strain either spontaneously or after induction. There was a close correlation between the lytic spectra of temperate and virulent phages. Among the strains studied, 25% were shown to be indicator strains. These results emphasized the high probability of development of temperate phages in a starter culture containing mesophilic lactic streptococci and therefore their importance as a cause of accidents in cheese making. PMID- 16345941 TI - Purification and Properties of a Malolactic Enzyme from a Strain of Leuconostoc mesenteroides Isolated from Grapes. AB - An enzymatic complex able to transform l-malate to l-lactate was obtained from a Leuconostoc mesenteroides strain isolated from grapes. The molecular weight was about 235,000, the isoelectric point was at pH 4.35, and the optimal pH for activity was 5.75. The malolactic activity followed a sequential pattern concerning the involved substrates. At pH values substantially different from the optimum, a positive cooperativity between malate molecules was observed. Oxamate, fructose-1, 6-diphosphate, and l-lactate acted as noncompetitive inhibitors, whereas succinate, citrate, and tartrate isomers produced a competitive inhibition. PMID- 16345942 TI - Characterization of Nocardia amarae as a Potent Biological Coalescing Agent of Water-Oil Emulsions. AB - Nocardia amarae grown in a liquid medium induced coalescence of emulsions which differed in type, composition of the organic phase, and structure of stabilizing emulsifiers. De-emulsifying activity varied with the type of growth medium, culture age, and postharvest treatment. Based on extraction and degradation studies, it was concluded that de-emulsifying properties are due to the bacterial cell surface. Thus, bacteria may provide a new source of de-emulsifying agents. PMID- 16345943 TI - Transformation of Adenine to 8-Hydroxyadenine by Strains of Oerskovia xanthineolytica. AB - The 8-hydroxy derivative of adenine (6-amino-1,7-dihydro-8H-purin-8-one) is produced from adenine by two Oerskovia xanthineolytica strains. This transformation by a microorganism has not been reported previously. No novel products of dissimilation of xanthine (3,7-dihydro-1H-purine-2,6-dione) or hypoxanthine (1,7-dihydro-6H-purin-6-one) were found. Xanthine was oxidized to uric acid, but intermediates in the breakdown of hypoxanthine could not be demonstrated. PMID- 16345944 TI - Actinomycetes and fungi in surface waters and in potable water. AB - In Finnish lakes and rivers used as water supplies, mesophilic fungi and actinomycetes were common, whereas thermophilic fungi and actinomycetes were present only in low concentrations. Fungi and actinomycetes were more abundant in eutrophic and mesotrophic lakes than in oligotrophic lakes. River water contained more thermophilic actinomycetes and fungi and mesophilic actinomycetes than did lake water. Runoff from soil seemed to be an important factor contributing to the incidence of these microbes in water. Chemical coagulation removed actinomycetes and fungi efficiently, but sand filtration allowed their passage. Disinfection could not prevent actinomycetes and fungi from reaching the distribution system. During infiltration in the production of recharged groundwater, mesophilic actinomycetes could even multiply appreciably. PMID- 16345945 TI - Seasonal variability and transport of suspended microfungi in a southeastern salt marsh. AB - Tidally induced fluctuations and transports of microfungi were investigated. Samples were collected at three depths from three stations positioned at a transect in a large salt marsh creek. Samples were taken every 1.5 h for 50 consecutive h during neap tides and 50 consecutive h during the corresponding spring tides. In each season, microfungi concentrations fluctuated out of phase with the tides during both neap and spring tides. Mean concentrations of suspended microfungi did not vary appreciably throughout the year. Fungi were exported from the marsh during the majority of the tidal cycles studied. The results suggest that microfungi may serve as indicators of water mass movements. PMID- 16345946 TI - Factors Affecting Yield and Safety of Protein Production from Cassava by Cephalosporium eichhorniae. AB - The properties of Cephalosporium eichhorniae 152 (ATCC 38255) affecting protein production from cassava carbohydrate, for use as an animal feed, were studied. This strain is a true thermophile, showing optimum growth at 45 degrees to 47 degrees C, maximum protein yield at 45 degrees C, and no growth at 25 degrees C. It has an optimum pH of about 3.8 and is obligately acidophilic, being unable to sustain growth at pH 6.0 and above in a liquid medium, or pH 7.0 and above on solid media. The optimum growth conditions of pH 3.8 and 45 degrees C were strongly inhibitive to potential contaminants. It rapidly hydrolyzed cassava starch. It did not utilize sucrose, but some (around 16%) of the small sucrose component of cassava was chemically hydrolyzed during the process. Growth with cassava meal (50 g/liter [circa 45 g/liter, glucose equivalent]) was complete in around 20 h, yielding around 22.5 g/liter (dry biomass), containing 41% crude protein (48 to 50% crude protein in the mycelium) and 31% true protein (7.0 g/liter). Resting and germinating spores (10 to 10 per animal) injected by various routes into normal and gamma-irradiated 6-week-old mice and 7-day-old chickens failed to initiate infections. PMID- 16345947 TI - Effects of plutonium on soil microorganisms. AB - As a first phase in an investigation of the role of the soil microflora in Pu complex formation and solubilization in soil, the effects of Pu concentration, form, and specific activity on microbial types, colony-forming units, and CO(2) evolution rate were determined in soils amended with C and N sources to optimize microbial activity. The effects of Pu differed with organism type and incubation time. After 30 days of incubation, aerobic sporeforming and anaerobic bacteria were significantly affected by soil Pu levels as low as 1 mug/g when Pu was added as the hydrolyzable Pu(NO(3))(4) (solubility, <0.1% in soil). Other classes of organisms, except the fungi, were significantly affected at soil Pu levels of 10 mug/g. Fungi were affected only at soil Pu levels of 180 mug/g. Soil CO(2) evolution rate and total accumulated CO(2) were affected by Pu only at the 180 mug/g level. Because of the possible role of resistant organisms in complex formation, the mechanisms of effects of Pu on the soil fungi were further evaluated. The effect of Pu on soil fungal colony-forming units was a function of Pu solubility in soil and Pu specific activity. When Pu was added in a soluble, complexed form [Pu(2)(diethylenetriaminepentaacetate)(3)], effects occurred at Pu levels of 1 mug/g and persisted for at least 95 days. Toxicity was due primarily to radiation effects rather than to chemical effects, suggesting that, at least in the case of the fungi, formation of Pu complexes would result primarily from ligands associated with normal (in contrast to chemically-induced) biochemical pathways. PMID- 16345948 TI - Differences among cowpea rhizobia in tolerance to high temperature and desiccation in soil. AB - Strains of cowpea rhizobia grew in mannitol-amended, nonsterile soil at 29 to 35 degrees C but not at 40 degrees C. Little decline in numbers of these bacteria occurred in dry, nonsterile soil incubated at 42 degrees C for 7 days. Strains of cowpea rhizobia differed widely in their tolerances to drying at 30 degrees C in nonsterile and sterile soil, and from less than 1 to 50% of the bacteria were still viable after 11 days. No relation was evident between tolerance to desiccation and the degree of aridity of the site from which the bacteria were isolated or their growth rates in culture, but strains not producing extracellular polysaccharide were often more tolerant than those producing extracellular polysaccharide. It is suggested that desiccation-tolerant rhizobia be used for the production of legume inoculants. PMID- 16345949 TI - Lignocellulolytic Enzymes Produced by Volvariella volvacea, the Edible Straw Mushroom. AB - Volvariella volvacea, commonly known as the straw or paddy mushroom, had the following growth characteristics: minimum temperature, 25 degrees C; optimal temperature, 37 degrees C; maximal temperature, 40 degrees C; pH optimum 6.0. Optimal pH for cellulase production was 5.5. The optimal initial pH for cellulase production and mycelial growth was found to be 6.0. The pH and temperature optima for cellulolytic activity were 5.0 and 50 degrees C, respectively. Maximal cellulolytic activity was obtained within 5 days in shake-flask culture. The cellulases were found to be partly cell free and partly cell bound during growth on microcrystalline cellulose. The endoglucanase activity was primarily extracellular, and beta-glucosidase activity was found exclusively extracellularly. Weak cellulase activity was detected when cells were grown on cellobiose and lactose. V. volvacea could not digest the lignin portion of newspaper in shake-flask cultivation. Phenol oxidase, an important enzyme in lignin biodegradation, also was lacking in the cell-free filtrate. However, the organism oxidized phenolic compounds when it was cultured on agar plates containing commercial lignin. PMID- 16345950 TI - Distribution of methanogenic and sulfate-reducing bacteria in near-shore marine sediments. AB - The distribution of methanogenic and sulfate-reducing bacteria was examined in sediments from three sites off the coast of eastern Connecticut and five sites in Long Island Sound. Both bacterial groups were detected at all sites. Three distributional patterns were observed: (i) four sites exhibited methanogenic and sulfate-reducing populations which were restricted to the upper 10 to 20 cm, with a predominance of sulfate reducers; (ii) three sites in western Long Island Sound exhibited a methanogenic population most abundant in sediments deeper than those occupied by sulfate reducers; (iii) at one site that was influenced by fresh groundwater, methanogens and sulfate reducers were numerous within the same depths; however, the number of sulfate reducers varied vertically and temporally with sulfate concentrations. It was concluded that the distributions of abundant methanogenic and sulfate-reducing bacteria were mutually exclusive. Methanogenic enrichments yielded all genera of methanogens except Methanosarcina, with the methanobacteria predominating. PMID- 16345951 TI - Rapid screen for bacteria degrading water-insoluble, solid hydrocarbons on agar plates. AB - A rapid procedure was devised for detecting on solid media bacteria able to degrade water-insoluble, solid hydrocarbons such as the polycyclic aromatic hydrocarbons phenanthrene, anthracene, and biphenyl. After Alcaligenes faecalis AFK2 was inoculated on a plate containing mineral salts agar, an ethereal solution of phenanthrene (about 10%, wt/vol) was sprayed on the surface of the plate, and the plate was incubated at 30 degrees C for 2 to 3 days. Colonies showing degradation were surrounded with clear zones on the opaque plate. A similar clear zone also was formed around colonies which had been grown on a succinate-mineral salts agar or nutrient agar, followed by spraying of the ethereal solution of phenanthrene and further incubating for 1 day. Other phenanthrene-assimilating bacteria, including Beijerinckia Bwt and Pseudomonas SPM64, also formed clear zones on phenanthrene-covered agar plates. This method was applicable to detection of bacteria able to assimilate anthracene, naphthalene, and biphenyl. PMID- 16345952 TI - Methanogenesis in big soda lake, nevada: an alkaline, moderately hypersaline desert lake. AB - Incubated sediment slurries from Big Soda Lake, Nevada, produced significant levels of CH(4), and production was inhibited by 2-bromoethanesulfonic acid and by autoclaving. Methane production was stimulated by methanol, trimethylamine, and, to a lesser extent, methionine. Surprisingly, hydrogen, acetate, and formate amendments provided only slight or no stimulation of methanogenesis. Methane production by sediment slurries had a pH optimum of 9.7. A methanol-grown enrichment culture containing a small, epifluorescent coccus as the predominant organism was recovered from sediments. The enrichment grew best when FeS or autoclaved sediment particles were included in the media, had a pH optimum of 9.7, and produced CH(4) from CH(3)OH. The methane formed by methanolgrown enrichment cultures was depleted in C by 72 to 77 per thousand relative to the methanol. PMID- 16345953 TI - The Microaerophile SPirillum volutans: Cultivation on Complex Liquid and Solid Media. AB - Spirillum volutans grows only under microaerobic conditions in a peptone succinate-salts broth, but can grow aerobically when the peptone is replaced by vitamin-free acid-hydrolyzed casein broth. The addition of potassium metabisulfite, norepinephrine, catalase or superoxide dismutase (SOD) permitted aerobic growth in peptone-succinate-salts broth. A combination of catalase and SOD had a synergistic effect. S. volutans lacked catalase and had only a low level of peroxidase activity, but did possess SOD activity (12 to 14 U/mg of protein). The organism was found to be extraordinarily sensitive to exogenous hydrogen peroxide. Illumination of peptone-succinate-salts broth generated hydrogen peroxide and rendered the medium inhibitory to growth. A combination of catalase and SOD prevented this inhibition. Growth of S. volutans on solid media, not previously possible, was accomplished by the use of vitamin-free acid hydrolyzed casein and peptone-succinate-salts agar media; maximum growth responses were dependent on the following combination of factors: addition of bisulfite, catalase, or SOD, protection of the media from illumination, incubation in a highly humid atmosphere, and incubation under atmospheres of 12% oxygen or less. The results indicate that the microaerophilic nature of S. volutans is attributable largely to the high sensitivity of the organism to exogenous hydrogen peroxide and, to a lesser extent, superoxide radicals occurring in the culture medium. PMID- 16345954 TI - Use of epifluorescence microscopy in studies of the germination and recovery of thermoactinomycetes. AB - Spore germination and growth of thermoactinomycetes were observed by epifluorescence microscopy. Each of the principal stages in germination was recognized and found to correspond to changes in phase-contrast appearance commonly monitored during endospore germination. The effects of novobiocin and nalidixic acid on germination of Thermoactinomyces vulgaris and T. thalpophilus were studied by using epifluorescence microscopy. Outgrowth but not initiation was inhibited by 100 mug of nalidixic acid per ml and 50 mug of novobiocin per ml. When each of the compounds, at various concentrations, was incorporated into a medium for thermoactinomycete recovery, the antibiotics were found to inhibit colony development. Samples of water and sediment incubated in a growth medium containing novobiocin and selective for thermoactinomycete species were examined by epifluorescence microscopy for total numbers of outgrown spores. Direct viable counts of outgrown spores indicated that the standard plate count enumerated less than 10% of the viable population of thermoactinomycetes. PMID- 16345955 TI - Simplified microassay for pullulan synthesis. AB - A simple radiometric microassay for extracellular polysaccharide elaboration by yeast-like cells of Aureobasidium pullulans was developed, based upon a procedure originally described by Catley (FEBS Lett. 20:174-176, 1972). Incorporation of [C]glucose into pullulan was linear with respect to time and cell dose. The pH and temperature optima for elaboration were 5.3 and 30 degrees C, respectively. Polysaccharide elaboration declined linearly with culture age. PMID- 16345956 TI - Effect of hard detergents on algae in a high-rate-oxidation pond. AB - Regular concentrations of hard detergents in domestic wastewater do not affect algal growth in a high-rate-oxidation pond. The addition of nonionic hard detergent at concentrations above 60 mg/liter decreased the algal concentration in the batch culture, and complete lysis of algal cells was observed within a few days at a detergent concentration of 100 mg/liter. PMID- 16345957 TI - Isolation of Clostridium thermocellum Auxotrophs. AB - Spontaneous and UV irradiation-induced auxotrophic mutants of Clostridium thermocellum, an anaerobic cellulolytic thermophile, were isolated after penicillin enrichment in a chemically defined medium. PMID- 16345959 TI - Nitrate accumulation in aerobic hypolimnia: relative importance of benthic and planktonic nitrifiers in an oligotrophic lake. AB - [This corrects the article on p. 566 in vol. 42.]. PMID- 16345958 TI - Influence of stabilizer concentration on effectiveness of chlorine as an algicide. AB - Cyanuric acid, used as chlorine stabilizer in swimming pool waters, has a relatively minor effect on the algicidal efficiency of free chlorine. The toxicity of free chlorine to three swimming pool algae was reduced slightly by 25 mg of cyanuric acid per liter if inhibiting, but less than algicidal, concentrations of chlorine were employed. Higher stabilizer concentrations (50, 100, and 200 mg/liter) generally resulted in no further reduction in the algicidal efficiency of free chlorine beyond that observed at 25 mg/liter. PMID- 16345960 TI - Denitrification in marine sediment: measurement of capacity and estimate of in situ rate. AB - The acetylene block technique was used to assay denitrification in undisturbed sediment cores of an intertidal mud flat. Nitrogen loss due to this process was estimated at 1 to 41 kg of N/hectare (ha) per year. Anaerobic nitrate-saturated slurry of the same sediment had a denitrification capacity of 1,026 kg of N/ha per year. The acetylene block technique failed at low nitrate concentrations, so that denitrification at average in situ nitrate concentrations could not be determined. Denitrification followed zero-order kinetics at nitrate concentrations high enough to allow successful blockage of N(2)O reduction. Thus, an estimate of in situ rates based on kinetic parameters and in situ nitrate concentrations was impossible. No denitrification was observed in a slurry of the top 1.5 cm under aerated conditions and nitrate saturation. In undisturbed sediment, significant denitrification occurred in few discrete sites within a matrix of undetectable or low activity. Despite numerous errors contributing to the uncertainty of the estimate of in situ rates, the result obtained by this method was considered more valuable than the determination of denitrification capacities. Methods which include severe changes of physical and chemical parameters may frequently lead to overestimates of denitrification rates. PMID- 16345961 TI - Production and loss of nitric oxide from denitrification in anaerobic brookston clay. AB - Nitric oxide, nitrous oxide, and nitrite ion production was measured in a Brookston clay column undergoing anaerobic denitrification. A flow system method was used whereby argon carrier gas continuously stripped soil gases from the column, allowing steady-state rates to be obtained. Over several days the temporal change in rates of these gases and NO(2) followed a pattern of increase and decay which may be expected of a reaction proceeding by several consecutive steps. The method permits observation of the relatively large net production rate of NO, which is normally not observed in static systems based on head space analysis of gaseous denitrification products. In the first several hours after the onset of anoxic conditions, the net rate of NO production, f(NO), increased sharply to a maximum ( approximately 1 x 10 mol of N/g of soil per min), paralleling the rapid increase in NO(2) level, and then followed a more gradual decline extending over approximately 45 h. A similar but less pronounced pattern was observed for N(2)O, with net rates of production being considerably less than for NO. The ratio [NO-N]/[N(2)O-N] decreased with time from approximately 2.5 at 6 h to approximately 2.0 at 45 h. Estimated rates of N(2) production appeared to be initially high, decreased rapidly within a few hours, and then gradually increased with time after the establishment of anaerobic conditions. PMID- 16345962 TI - Apparent and measured rates of nitrification in the hypolimnion of a mesotrophic lake. AB - Three distinct phases were observed in the change of dissolved inorganic nitrogen concentrations in the hypolimnion of Grasmere. The second phase of decreasing ammonia and increasing nitrate concentrations was typical of the nitrification process. Observations on nitrate concentration gradients between surface sediments and the water column and experiments using the nitrification inhibitor N-Serve indicated the in situ activity of chemolithotrophic nitrifying organisms. Nitrification rates were estimated throughout the period of stratification by using the N-Serve and [C]bicarbonate uptake method. Comparison of the field nitrate concentrations with the predicted nitrate concentrations (from estimates of the nitrification rate) indicated that the method underestimated the true rate of nitrification. Possible reasons for this are discussed. PMID- 16345963 TI - Intermediary metabolism of organic matter in the sediments of a eutrophic lake. AB - The rates, products, and controls of the metabolism of fermentation intermediates in the sediments of a eutrophic lake were examined. C-fatty acids were directly injected into sediment subcores for turnover rate measurements. The highest rates of acetate turnover were in surface sediments (0- to 2-cm depth). Methane was the dominant product of acetate metabolism at all depths. Simultaneous measurements of acetate, propionate, and lactate turnover in surface sediments gave turnover rates of 159, 20, and 3 muM/h, respectively. [2-C]propionate and [U-C]lactate were metabolized to [C]acetate, CO(2), and CH(4). [C]formate was completely converted to CO(2) in less than 1 min. Inhibition of methanogenesis with chloroform resulted in an immediate accumulation of volatile fatty acids and hydrogen. Hydrogen inhibited the metabolism of C(3)-C(5) volatile fatty acids. The rates of fatty acid production were estimated from the rates of fatty acid accumulation in the presence of chloroform or hydrogen. The mean molar rates of production were acetate, 82%; propionate, 13%; butyrates, 2%; and valerates, 3%. A working model for carbon and electron flow is presented which illustrates that fermentation and methanogenesis are the predominate steps in carbon flow and that there is a close interaction between fermentative bacteria, acetogenic hydrogen producing bacteria, and methanogens. PMID- 16345964 TI - Isolation and characterization of ultramicrobacteria from a gulf coast estuary. AB - Viable bacteria were recovered from estuarine waters passed through a 0.2-mum polycarbonate membrane filter. The recovery method included the use of a dilute nutrient broth for primary enrichment followed by conditioning of the organism to a dilute nutrient solid medium. These bacteria were gram-negative rods and coccobacilli having an NaCl requirement and, upon initial culturing, low nutritional requirements. In response to increased nutrient preparations, these microorganisms underwent an increase in size and growth rate, giving rise to visible colonies. Phenotypic characterization suggests that species of Vibrio, Aeromonas, Pseudomonas, and Alcaligenes were among the isolates. The abundance and the nutritional requirements of these ultramicrobacteria imply that they represent a class of microorganisms which have successfully adjusted to poor nutrient conditions. PMID- 16345965 TI - Transformation of 25-Hydroxycholesterol by Rhizoctonia muneratii. AB - Rhizoctonia muneratii ATCC 13247 was capable of catalyzing the conversion of 25 hydroxycholesterol to the 3-oxo-4-ene. Hydroxylation at the 6alpha and 6beta positions with the concomitant 3-oxo-4-ene transformation was also observed. The addition of Triton X-100 to the growth medium immediately before steroid addition selectively prevented the appearance of the respective 6alpha-hydroxylated products. The observed conversions were indicative of the utility of microbial transformations in preparing hydroxylated steroids which can be useful for further synthetic modification or for study as pharmacological agents. PMID- 16345966 TI - Survival of cowpea rhizobia in soil as affected by soil temperature and moisture. AB - Successful inoculation of peanuts and cowpeas depends on the survival of rhizobia in soils which fluctuate between wide temperature and moisture extremes. Survival of two cowpea rhizobial strains (TAL309 and 3281) and two peanut rhizobial strains (T-1 and 201) was measured in two soils under three moisture conditions (air-dry, moist (-0.33 bar), and saturated soil) and at two temperatures (25 and 35 degrees C) when soil was not sterilized and at 40 degrees C when soil was sterilized. Populations of rhizobia were measured periodically for 45 days. The results in nonsterilized soil indicated that strain 201 survived relatively well under all environmental conditions. The 35 degrees C temperature in conjunction with the air-dry or saturated soil was the most detrimental to survival. At this temperature, the numbers of strains T-1, TAL309, and 3281 decreased about 2 logs in dry soil and 2.5 logs in saturated soil during 45 days of incubation. In sterilized soil, the populations of all strains in moist soil increased during the first 2 weeks, but decreased rapidly when incubated under dry conditions. The populations did not decline under saturated soil conditions. From these results it appears that rhizobial strains to be used for inoculant production should be screened under simulated field conditions for enhanced survival before their selection for commercial inoculant production. PMID- 16345967 TI - Aminopeptidase profiles of four species of phytophthora. AB - Phytophthora palmivora, P. cinnamomi, P. drechsleri, and P. cactorum were readily separated on the basis of the aminopeptidase substrate specificities of their mycelial suspensions. Variability among isolates of the same species was not significant with most substrates and isolates tested, regardless of source. Both qualitative and quantitative differences in enzyme activity were useful for species identification. Differentiation of these four species was possible with comparative reactions of l-alanyl-, l-arginyl-, l-benzoylarginyl, l-gamma glutamyl-, l-glycyl-, l-hydroxyprolyl-, l-leucyl-, l-lysyl-, 4-methoxyleucyl-, l prolyl-, and 4-methoxyalanyl-beta-naphthylamides. Variation in peptidase activity was usually detectable after 4 h of incubation, with increased activity sometimes manifest after 24 h of incubation. P. palmivora exhibited the lowest, and P. drechsleri exhibited the highest, overall peptidase activity. This fluorescent aminopeptidase profile procedure provides a relatively rapid method to supplement other taxonomic criteria for identification of these species of Phytophthora. PMID- 16345968 TI - Biodegradation of [C]Tri-p-Cresyl Phosphate in a Laboratory Activated-Sludge System. AB - Biodegradation of [C]tri-p-cresyl phosphate was studied in a laboratory model sewage treatment system to develop information on the nature of its transformation products. In 24-h experiments, 70 to 80% of tri-p-cresyl phosphate added at the 1-mug/ml level was degraded. The remaining parent compound was associated with the sludge solids. The major metabolite extracted with ethyl ether from the aqueous phase was identified as p-hydroxybenzoic acid by gas chromatography-mass spectrometry. Two unstable ether-extractable metabolites were not identified. The half-life of [C]tri-p-cresyl phosphate was estimated to be 7.5 h. PMID- 16345969 TI - Nodulation efficiency of legume inoculation as determined by intrinsic antibiotic resistance. AB - Patterns of intrinsic resistance and susceptibility to different levels of antibiotics were determined for strains of both fast- and slow-growing rhizobia. These patterns were stable to plant passage when they were used to identify Rhizobium strains in nodule suspensions or nodule isolates. The method of identification by intrinsic resistance and susceptibility patterns was reliable for identifying strains in field nodules when strains were first isolated from the nodules to provide a standard inoculum size and then typed on antibiotic containing media. Other patterns of resistance were encountered during identification of field isolates; these patterns may have resulted from acquired resistance to certain antibiotics or from mixed infections of the nodules. The occurrence of resistance patterns identical to those of inoculant strains among native strains was directly related to the size of the soil population. High strain recovery was associated directly with high rates of inoculation. PMID- 16345970 TI - Single-Cell Protein Production by the Acid-Tolerant Fungus Scytalidium acidophilum from Acid Hydrolysates of Waste Paper. AB - The bioconversion of waste paper to single-cell protein at pH <1 by Scytalidium acidophilum is described. Waste paper pretreated with 72% H(2)SO(4) at 4 degrees C was diluted with water to a pH of <0.1 and hydrolyzed. This yielded an adequate sugar-containing substrate for the growth of the fungus. A total of 97% of the sugars (glucose, galactose, mannose, xylose, arabinose) in the hydrolysates were converted to cell biomass. Microbial contamination was not observed. Based on the sugars consumed, S. acidophilum produced higher yields in shake cultures than many other Fungi Imperfecti. In aerated cultures, productivity increased, and yields of 43 to 46% containing 44 to 47% crude protein were obtained. This compares favorably with Candida utilis, a yeast used commercially to produce single-cell protein. The chemical constituents and the essential amino acids of the fungal cells were similar to those of other fungi. The nucleic acid content was characteristic of microbes containing low levels of nucleic acid. The advantages of using S. acidophilum for single-cell protein production are discussed. PMID- 16345971 TI - Denitrification, nitrate reduction, and oxygen consumption in coastal and estuarine sediments. AB - Denitrification and consumption of oxygen and nitrate in sediments from Tama Estuary, Odawa Bay, and Tokyo Bay were measured in an experimental sediment-water system. Filtered seawater containing [N]nitrate flowed continuously over undisturbed sediments, and the concentrations of O(2), nitrate, and nitrite in the influent and effluent and of N(2) in the effluent were monitored. Under steady-state conditions, the rate of nitrate consumption was the same order of magnitude as the rate of oxygen consumption in Tama Estuary sediments, whereas the former rate was one order of magnitude lower than the latter rate in Odawa Bay and Tokyo Bay sediments. Denitrification accounted for 27 to 57% of the nitrate consumption. PMID- 16345972 TI - Characteristics of Protease Production by Cephalosporium sp. AB - We investigated protease formation by Cephalosporium sp. strain KM388, which produced trypsin inhibitor in the same cultures, in medium containing polypeptone, meat extract, and glucose (natural medium) and in medium containing NaNO(3), glucose, and yeast extract (semisynthetic medium). In natural medium, protease was secreted into the culture broth after cessation of growth caused by consumption of the polypeptone, the growth-limiting substrate. Enzyme formation in the stationary growth phase was due to de novo and so-called preferential synthesis, because cycloheximide immediately inhibited enzyme formation. In semisynthetic medium, protease was produced in parallel with mycelial growth, but production was repressed by the addition of polypeptone to the medium; protease production began after the added polypeptone was consumed. On the other hand, if glucose was eliminated from natural medium, the lag period of initiation of enzyme production was reduced until the late exponential phase. The addition of phosphate up to a concentration of 1.0% to natural medium also shortened the lag period and damped the pH change of the broth during cultivation. PMID- 16345973 TI - Impact of mount st. Helens eruption on bacteriology of lakes in the blast zone. AB - Lakes lying within the blast zone of Mount St. Helens showed dramatic increases in heterotrophic bacterial numbers after the eruption of 18 May 1980. The total microscopic counts of bacteria in some of the most severely affected lakes were more than 10 cells per ml, an order of magnitude above the counts in outlying control lakes. Likewise, the numbers of viable bacteria reached levels of more than 10 cells per ml, compated with fewer than 10 cells per ml in control lakes. The CPS medium used for enumeration provided growth of up to 81.5% of the bacteria during sampling of one of the blast zone lakes. The high numbers of bacteria and the efficacy of the viable enumeration procedure are evidence that the lakes have been transformed rapidly from oligotrophy to eutrophy due to the eruption and its aftermath. Organic material leached from the devastated forest vegetation is thought to be responsible for the enrichment of heterotrophs. Total coliform bacteria were found in all of the blast zone lakes, and some lakes contained fecal coliform bacteria. Klebsiella pneumoniae was the predominant total coliform and was also identified as one of the fecal coliform bacteria, although Escherichia coli was the predominant species in that category. Our data indicate that bacterial populations peaked in the outer blast zone lakes in the summer of 1980 and in most of the inner lakes during the summer of 1981. PMID- 16345974 TI - Production of Extracellular Pigment by a Mutant of Monascus kaoliang sp. nov. AB - A hyperpigment-producing mutant, R-10847, was derived from Monascus kaoliang F-2 (ATCC 26264) through a series of mutagenesis steps. The mutant produced a large quantity of Monascus pigment when grown in mantou (steamed bread) by solid culture. The mutant produced pigments extracellularly by extruding the pigments outside the cell in a lump together with some viscous substances. The productivity of pigment was about 100-fold greater than that of the wild type. The mutant lost the capability of spore formation, the growth rate decreased, and both the size and quantity of conidia were reduced. The color of the pigment produced by the mutant changed from orange to deep red. PMID- 16345975 TI - Recognition of leguminous hosts by a promiscuous Rhizobium strain. AB - The lima bean (Phaseolus lunatus L.) and the pole bean (Phaseolus vulgaris L.) are nodulated by rhizobia of two different cross-inoculation groups. Rhizobium sp. 127E15, a cowpea-type Rhizobium, can induce effective nodules on the lima bean and partially effective nodules on the pole bean. Rhizobium phaseoli 127K14 can induce effective nodules on the pole bean but does not reciprocally nodulate the lima bean. Root hairs of the lima bean when inoculated with Rhizobium sp. 127E15 showed tip curling and swelling and infection thread formation as observed by light microscopy and scanning electron microscopy. When lima bean root hairs were inoculated with R. phaseoli 127K14, no host-specific responses were observed. Pole bean root hairs that had been inoculated with R. phaseoli 127K14 or Rhizobium sp. 127E15 also showed tip curling and swelling and infection thread formation. Colonization of lima bean root hairs by Rhizobium sp. 127E15 and pole bean root hairs by R. phaseoli 127K14 or Rhizobium sp. 127E15 appeared to involve the elaboration of microfibrils. This study showed that when Rhizobium sp. 127E15 nodulates a host of a different cross-inoculation group, it elicits the same specific host responses as it does from a host of the same cross-inoculation group. PMID- 16345976 TI - Methods for studying aquatic bacteriophage ecology. AB - Methods are described for enumerating the different kinds of bacteriophage present in virus concentrates prepared from 120 liters of water. Although developed specifically for use with coliphages, they should be applicable to viruses infecting other hosts. These methods involve mixed indicators, equilibrium buoyant-density centrifugation, use of enzymes or inhibitors or both, and specific hybridization probes, either alone or in combination. With these methods, it was possible to specifically enumerate filamentous and isometric male specific (F-specific) phages, the different classes of P-group plasmid-specific phages, phiX174-like phages, and lambda-like phages. Some applications of these methods, including measurement of virus inactivation in the environment and the extent of fecal pollution, are discussed. PMID- 16345977 TI - Microbial dynamics of an epilithic mat community in a high alpine stream. AB - Studies were conducted to examine interrelationships between the heterotrophic and phototrophic populations within an epilithic community in the outlet stream of a high alpine lake. Levels of nitrates, phosphates, and total organic compounds in the lake were consistently near the lower limits of detectability. Microscopic examination of the community by phase-contrast light microscopy and scanning electron microscopy revealed diatoms, filamentous algae, and bacteria embedded within a dense gelatinous matrix. Chlorophyll a and primary productivity measurements had peak values in early August, with subsequent declines. Bacterial heterotrophic activity, as measured by V(max), turnover rate, and relative activity, increased significantly as the phototrophic community declined. This trend in heterotrophic activity was not accompanied by an increase in total bacterial numbers as determined by epi-illuminated fluorescence microscopy. These results suggest that the phototrophic community responded to changes in, or interactions among, various chemical and physical factors throughout the study period. The catabolic activity of the sessile bacteria appeared to be positively influenced by changes in the mat environment resulting from the decline of the phototrophic populations. PMID- 16345978 TI - Geosmin and 2-methylisoborneol from cyanobacteria in three water supply systems. AB - Three Oscillatoria strains and one Anabaena species were isolated from three different water supply systems in California that experienced earthy-musty taste and odor problems in their drinking water. Unialgal cultures, free of actinomycetes, were purged using the Grob closed-loop stripping analysis method, and the resulting methylene chloride extracts were analyzed on a gas chromatograph/mass spectrometer. Geosmin was produced by Oscillatoria simplicissima and Anabaena scheremetievi, and 2-methylisoborneol was produced by O. curviceps and O. tenuis. These compounds are the two major causes of earthy musty tastes and odors in water. In three instances, the major odorant found in culture was previously identified in the water or sediment sample from which the respective organism was isolated. O. curviceps was implicated in a taste and odor episode involving 2-methylisoborneol in a major reservoir. Geosmin and 2 methylisoborneol were easily detected with culture samples of only 4 to 25 ml. PMID- 16345979 TI - Effect of Several Algicides and Surfactants on the Filamentous Bacterium Leucothrix mucor Oersted. AB - Leucothrix mucor Oersted, the primary cause of filamentous gill disease in cultured shrimp, was treated with algicides and surfactants by a serial dilution method. It was found to be highly sensitive to those algicides containing quaternary ammonium compounds. Copper-containing compounds and Formalin were less effective, whereas potassium permanganate and two surfactants, Dreft and Alconox, were ineffective. PMID- 16345980 TI - Clostridium acetobutylicum Protoplast Formation and Regeneration. AB - Techniques and media for the production and regeneration of stable Clostridium acetobutylicum protoplasts are described. PMID- 16345981 TI - Influence of acetylene on growth of sulfate-respiring bacteria. AB - At a concentration of 20% of the atmosphere of the culture flasks, acetylene inhibited growth and carbon dioxide production by Desulfovibrio desulfuricans and Desulfovibrio gigas. The bacteria did not reduce acetylene to ethylene, and neither acetylene dicarboxylic acid nor ethylene was inhibitory. At 10%, acetylene was partially inhibitory for the desulfovibrios. At 5%, acetylene impeded the rate but did not limit the extent of growth and catabolism of the desulfovibrios. Desulfotomaculum ruminis was affected only negligibly, if at all, by acetylene and ethylene at any of these concentrations. PMID- 16345982 TI - Isolation and Characterization of Two New Trichothecenes from Fusarium sporotrichioides Strain M-1-1. AB - [This corrects the article on p. 542 in vol. 42.]. PMID- 16345983 TI - Free ammonia inhibition of algal photosynthesis in intensive cultures. AB - The effect of free NH(3) inhibition on short-term photosynthesis was investigated in three microalgal species: the freshwater chlorophyte Scenedesmus obliquus, the marine diatom Phaeodactylum tricornutum and the marine chlorophyte Dunaliella tertiolecta. By performing a series of assays at various concentrations of added NH(4)Cl and culture pH, we demonstrated that the inhibitory compound was free NH(3) and that pH played no role in determining the magnitude of inhibition, other than in establishing the degree of dissociation of nontoxic NH(4) to toxic NH(3). When corrections were made for pH, all three species displayed the same sigmoidal response curve to free NH(3) concentration; 1.2 mM NH(3) led to 50% reduction in photoassimilation of C. Based on literature values, some marine phytoplankton appear to be significantly more sensitive to free NH(3) than were the test species, which are noted for their excellent growth characteristics. However, the combination of low algal biomass and strong pH buffering commonly found in most marine and many freshwater environments probably limits the possibilities for NH(3) toxicity to low alkalinity freshwaters and intensive algal cultures in which NH(4) is the main source of N. Such conditions occur commonly in algal wastewater treatment systems. PMID- 16345984 TI - Interactions of bacterial and amoebal populations in soil microcosms with fluctuating moisture content. AB - Sterilized soil samples (20 g of soil per 50-ml flask), amended with 600 mug of glucose-carbon and 60 mug of NH(4)-N . g of dry soil, were inoculated with bacteria (Pseudomonas paucimobilis) alone or with bacteria and amoebae (Acanthamoeba polyphaga). We used wet-dry treatments, which involved air drying the samples to a moisture content of approximately 2% and remoistening the samples three times during the 83-day experiment. Control treatments were kept moist. In the absence of amoebae, bacterial populations were reduced by the first drying to about 60% of the moist control populations, but the third drying had no such effect. With amoebae present, bacterial numbers were not significantly affected by the dryings. Amoebal grazing reduced bacterial populations to 20 to 25% of the ungrazed bacterial populations in both moisture treatments. Encystment was an efficient survival mechanism for amoebae subjected to wet-dry cycles. The amoebal population was entirely encysted in dry soil, but the total number of amoebae was not affected by the three dryings. Growth efficiencies for amoebae feeding on bacteria were 0.33 and 0.39 for wet-dry and constantly moist treatments, respectively, results that compared well with those previously reported for Acanthamoeba spp. PMID- 16345985 TI - Effect of sodium chloride on bakers' yeast growing in gelatin. AB - In recent years, industrial fermentation researchers have shifted their attention from liquid to solid and semisolid culture conditions. We converted liquid cultures to the semisolid mode by adding high levels of gelatin. Previous studies on liquid cultures have revealed the inhibitory activity of mineral salts, such as NaCl, on the fermentation of sugars by yeasts. We made a kinetic study of the effects of 1 to 5% (wt/vol) NaCl on the alcoholic fermentations of glucose by Saccharomyces cerevisiae in a growth medium containing 16% gelatin. Our results showed that the effect of high salt content on semisolid culture is essentially the same as the effect on liquid culture; i.e., as the salt content increased, the following occurred: (i) the growth of yeasts decreased, (ii) the lag period of the yeast biomass curve lengthened, (iii) the sugar intake was lowered, (iv) the yield of ethanol was reduced, and (v) the production of glycerol was increased. We observed a new relationship correlating the area of kinetic hysteresis with ethanol production rate, acetaldehyde concentration, and the initial NaCl concentration. PMID- 16345986 TI - Relative Efficacy of Different Alfalfa Cultivar-Rhizobium meliloti Strain Combinations for Symbiotic Nitrogen Fixation. AB - Five Rhizobium meliloti isolates known to have different capabilities for expression of nitrogenase activity under symbiotic conditions were used to inoculate four representative Medicago sativa cultivars under aseptic conditions. Nitrogenase activities and respiratory activity were measured for whole plants and excised nodules. Dry weights and nodule numbers were also recorded after 4 weeks of growth in plastic pouches on a nitrogen-free nutrient medium. Hydrogen evolution and acetylene reduction rates were used to calculate the fraction of reducing power allocated to dinitrogen reduction. Although the efficiency of the system defined in this way was poorly correlated with plant yield, a very high linear correlation was obtained between yield and the algebraic product of nitrogenase activity and efficiency. High correlation (r > 0.78) was obtained between respiration and nitrogenase activity for whole plants as well as for excised nodules. Nodular respiration accounted for between 10 and 20% of the total plant dark respiration. The four test cultivars exhibited significantly different symbiotic responses to the inocula, although trends in potential for expression of the nitrogenase system by the five R. meliloti strains were evident. There was significant interaction between the host plant and symbiont in determining nitrogenase activity and yield. This screening method allows quantitative discrimination between effective and ineffective host-inoculum combinations. PMID- 16345987 TI - Production and vertical flux of attached bacteria in the hudson river plume of the new york bight as studied with floating sediment traps. AB - We investigated the growth and vertical flux of attached bacteria with floating sediment traps in the Hudson River Plume of the New York Bight during the spring diatom blooms. Traps were floated at the base of the mixed layer (ca. 10 m) for 1 day periods. After recovery, we measured bacterial abundance and rates of [methyl H]thymidine incorporation in the trap samples. The vertical flux of attached bacteria was estimated with a model formulated to distinguish between bacterial accumulation in traps due to in situ growth and that due to vertical flux. Attached bacterial flux ranged from 0.6 x 10 to 2.0 x 10 cells m day, and attached bacterial settling rates of 0.1 to 1.0 m day were observed during periods of vertical particulate organic carbon flux ranging from 254 to 1,267 mg of C m day. In situ growth of bacteria in sediment traps was unimportant as a source of bacterial increase when compared with vertical flux during our study. The vertical flux of attached bacteria removed 3 to 67% of the total daily bacterial production from the water column. Particulate organic carbon is not significantly mineralized by attached bacteria during its descent to the sea floor in the plume area during this period, when water temperature and grazing rates are at their annual minima. PMID- 16345988 TI - Ametryne and prometryne as sulfur sources for bacteria. AB - Bacteria were isolated that could utilize quantitatively the s-triazine herbicide prometryne [N,N' -bis(1-methylethyl)-6-(methylthio)-1,3,5-triazine-2,4-diamine] or ametryne [N-ethyl-N'-(1-methylethyl)-6-(methylthio)-1,3,5-triazine- 2,4 diamine], or both, as a sole source of sulfur for growth. The success of enrichments depended on previous exposure of the soil inoculum to s-triazine herbicides. Deaminoethylametryne [4-(1-methylethyl)amino-6-(methylthio)-1,3,5 triazine-2-(1H)-one], methylsulfonic acid, and sodium sulfate could also be used as sulfur sources. Utilization of a compound was quantified as the growth yield per mole of sulfur supplied. Yields were about 6 kg of protein per mol of sulfur. The product of the desulfuration of an s-triazine was identified as the corresponding hydroxy-derivative. This is the first substantiated report of the utilization of these s-triazines as sulfur sources by bacteria. PMID- 16345989 TI - Legume-Rhizobium interactions: cowpea root exudate elicits faster nodulation response by Rhizobium species. AB - Preinfection events in legume-Rhizobium symbiosis were analyzed by studying the different nodulation behaviors of two rhizobial strains in cowpeas (Vigna sinensis). Log-phase cultures of Rhizobium sp. strain 1001, an isolate from the plant nodule, initiated host responses leading to infection within 2 h after inoculation, whereas log-phase cultures of Rhizobium sp. strain 32H1 took at least 7 h to trigger a discernible response. The delay observed with strain 32H1 could be eliminated by incubating the rhizobial suspension, before inoculation, for 4.5 h either in the cowpea rhizosphere/rhizoplane condition or in the root exudate of cowpea plants, grown without NH(4) in the rooting medium. The delay could not be eliminated by incubating the rhizobial suspension in the rooting medium of plants grown in the presence of 5 mM NH(4), indicating that there is a regulatory role of combined nitrogen in triggering preinfection events by the legume. The substance(s) in the root exudate which elicited the faster nodulation response by Rhizobium sp. strain 32H1 could be separated into a high-molecular weight fraction by Sephadex G-100 gel filtration. The data support the notion that legume roots release substances that favor the development of rhizobial features essential for infection and nodulation. PMID- 16345990 TI - Respiration of tritiated substrates in heterotrophic activity assays. AB - The respiration percentage of H-labeled glucose by heterotrophic microorganisms in water samples was determined by a method based on equimolar addition of [H]- and [C]glucose to samples. The respiration percentage of [H]glucose exceeded that of [C]glucose by 10% over the assayed range of respiration rates. Respiration percentages diminished with decreasing temperature, the relation between respiration of [H]- and [C]glucose remaining the same. The significance of tritiated water respiration with regard to experimental design is emphasized. PMID- 16345991 TI - Production of Plant Growth-Regulating Substances by the Vesicular-Arbuscular Mycorrhizal Fungus Glomus mosseae. AB - Glomus mosseae, a representative species of Endogonaceae (Phycomycetes) able to form vesicular-arbuscular mycorrhiza, was investigated for phytohormone production. Spores of G. mosseae were axenically germinated in water, and the resultant mycelial growth was assayed by standard procedures for extracting plant hormones from microbial cultures. Paper partition chromatography and specific bioassays were used to separate and identify plant growth-regulating substances. The microorganism synthesized at least two gibberellin-like substances, one with R(f) corresponding in position to authentic gibberellic acid, and four substances with the properties of cytokinins. PMID- 16345992 TI - Nitrogen fixation (acetylene reduction) associated with duckweed (lemnaceae) mats. AB - Duckweed (Lemnaceae) mats in Texas and Florida were investigated, using the acetylene reduction assay, to determine whether nitrogen fixation occurred in these floating aquatic macrophyte communities. N(2)-fixing microorganisms were enumerated by plating or most-probable-number techniques, using appropriate N free media. Results of the investigations indicated that substantial N(2) fixation (C(2)H(2)) was associated with duckweed mats in Texas and Florida. Acetylene reduction values ranged from 1 to 18 mumol of C(2)H(4) g (dry weight) day for samples incubated aerobically in light. Dark N(2) fixation was always two to fivefold lower. 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (7 to 10 muM) reduced acetylene reduction to levels intermediate between light and dark incubation. Acetylene reduction was generally greatest for samples incubated anaerobically in the light. It was estimated that 15 to 20% of the N requirement of the duckweed could be supplied through biological nitrogen fixation. N(2)-fixing heterotrophic bacteria (10 cells g [wet weight] and cyanobacteria (10 propagules g [wet weight] were associated with the duckweed mats. Azotobacter sp. was not detected in these investigations. One diazotrophic isolate was classified as Klebsiella. PMID- 16345993 TI - Enhancement by Sodium Dodecyl Sulfate of Pigment Formation in Serratia marcescens O8. AB - Three methods were used to determine the enhancement by sodium dodecyl sulfate (SDS) of prodigiosin formation in Serratia marcescens O8. The results of the agar disk diffusion method indicated that pigment formation was dependent upon the concentration of SDS. Diameters of the pigment zones were proportional to the logarithm of SDS concentrations of 300 to 1,500 mug/ml. When bacteria were grown in broth containing SDS from 0 to 800 mug/ml and the pigment extracts were analyzed spectrophotometrically, a similar enhancement of pigment formation was observed. Finally, these results were confirmed by high-performance liquid chromatographic analysis of the extracts. Prodigiosin appeared to be the sole component with increased synthesis. The possible mechanism of the SDS enhancement effect could be explained by an increase in negative binding sites by the association of SDS with a cell envelope component(s). These binding sites may be required for prodigiosin synthesis. PMID- 16345994 TI - Dissimilatory Reduction of NO(2) to NH(4) and N(2)O by a Soil Citrobacter sp. AB - Dissimilatory reduction of NO(2) to N(2)O and NH(4) by a soil Citrobacter sp. was studied in an attempt to elucidate the physiological and ecological significance of N(2)O production by this mechanism. In batch cultures with defined media, NO(2) reduction to NH(4) was favored by high glucose and low NO(3) concentrations. Nitrous oxide production was greatest at high glucose and intermediate NO(3) concentrations. With succinate as the energy source, little or no NO(2) was reduced to NH(4) but N(2)O was produced. Resting cell suspensions reduced NO(2) simultaneously to N(2)O and free extracellular NH(4). Chloramphenicol prevented the induction of N(2)O-producing activity. The K(m) for NO(2) reduction to N(2)O was estimated to be 0.9 mM NO(2), yet the apparent K(m) for overall NO(2) reduction was considerably lower, no greater than 0.04 mM NO(2). Activities for N(2)O and NH(4) production increased markedly after depletion of NO(3) from the media. Amendment with NO(3) inhibited N(2)O and NH(4) production by molybdate-grown cells but not by tungstate-grown cells. Sulfite inhibited production of NH(4) but not of N(2)O. In a related experiment, three Escherichia coli mutants lacking NADH-dependent nitrite reductase produced N(2)O at rates equal to the wild type. These observations suggest that N(2)O is produced enzymatically but not by the same enzyme system responsible for dissimilatory reduction of NO(2) to NH(4). PMID- 16345995 TI - Uptake of Cyclic AMP by Natural Populations of Marine Bacteria. AB - The major objective of this study was to describe the mechanism(s) of cyclic AMP uptake by natural populations of marine bacteria. A second objective was to determine whether this uptake could contribute to the intracellular regulatory pool of cyclic AMP. Using high-specific-activity P-labeled cyclic AMP, we found several high-affinity uptake systems. The highest-affinity system had a half saturation constant of <10 pM. This system was extremely specific for cyclic nucleotides, particularly cyclic AMP. It appeared to meet the criteria for active transport. Uptake of cyclic AMP over a wide concentration range (up to 2 muM) showed multiphasic kinetics, with half-saturation constants of 1 nM and greater. These lower-affinity systems were much less specific for cyclic nucleotides. Although much of the labeled cyclic AMP taken up by the high-affinity systems was metabolized, some remained as intact cyclic AMP within the cells during 1 h of incubation. This suggests that at least some of the bacteria use cyclic AMP dissolved in seawater to augment their intracellular pools. PMID- 16345996 TI - Iron Oxidation and Precipitation of Ferric Hydroxysulfates by Resting Thiobacillus ferrooxidans Cells. AB - The oxidation of ferrous ions, in acid solution, by resting suspensions of Thiobacillus ferrooxidans produced sediments consisting of crystalline jarosites, amorphous ferric hydroxysulfates, or both. These products differed conspicuously in chemical composition and infrared spectra from precipitates formed by abiotic oxidation under similar conditions. The amorphous sediments, produced by bacterial oxidation, exhibited a distinctive fibroporous microstructure when examined by scanning electron microscopy. Infrared spectra indicated outer-sphere coordination of Fe(III) by sulfate ions, as well as inner-sphere coordination by water molecules and bridging hydroxo groups. In the presence of excess sulfate and appropriate monovalent cations, jarosites, instead of amorphous ferric hydroxysulfates, precipitated from bacterially oxidized iron solutions. It is proposed that the jarositic precipitates result from the conversion of outer sphere (T(d)) sulfate, present in a soluble polymeric Fe(III) complex, to inner sphere (C(3v)) bridging sulfate. The amorphous precipitates result from the further polymerization of hydroxo-linked iron octahedra and charge stabilized aggregation of the resulting iron complexes in solution. This view was supported by observations that bacterially oxidized iron solutions gave rise to either amorphous or jarositic sediments in response to ionic environments imposed after oxidation had been completed and the bacteria had been removed by filtration. PMID- 16345997 TI - Use of hoechst dyes 33258 and 33342 for enumeration of attached and planktonic bacteria. AB - The DNA-specific fluorochromes Hoechst 33258 and 33342 were used to enumerate aquatic bacteria by epifluorescent direct counts. Cultures of estuarine bacteria gave identical counts when stained with Hoechst 33258 or acridine orange, whereas natural populations of aquatic bacteria gave 92 to 98.5% of the acridine orange counts. The technique had distinct advantages over acridine orange when enumerating bacteria on surfaces which bind acridine orange, such as polystyrene. PMID- 16345998 TI - Use of nitrifier activity measurements to estimate the efficiency of viable nitrifier counts in soils and sediments. AB - A procedure for estimating the efficiency of the most-probable number (MPN) technique for counting ammonium-oxidizing bacteria was tested on sediments and soils collected from Delaware Inlet, Nelson, New Zealand. The procedure involved estimating the nitrifier populations required to produce observed activities and comparing these estimates with the MPN-countable populations. MPN counts ranged between 0.15 x 10 to 3.0 x 10 cells g in sediments and between 4.4 x 10 to 19 x 10 cells g in soils. These counts were only 0.1 to 5.0% of the estimated populations that would be required to produce the observed activity. Similar efficiency calculations were made for data already in the literature, and these calculations gave much higher percentages. Thus, we concluded that for the soils and sediments we studied, the MPN counting technique greatly underestimated the populations present and that the efficiency calculation could be used as a counting efficiency index. PMID- 16346000 TI - Isolation of a Strain of Clostridium thermoaceticum Capable of Growth and Acetic Acid Production at pH 4.5. AB - [This corrects the article on p. 118 in vol. 43.]. PMID- 16345999 TI - Tetrazolium reduction-malachite green method for assessing the viability of filamentous bacteria in activated sludge. AB - A method was developed to assess the activity of filamentous bacteria in activated sludge. It involves the incubation of activated sludge with 2(p iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride followed by staining with malachite green. Both cells and 2(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride-formazan crystals can be observed in prepared specimens by using bright-field microscopy. This procedure allowed us to distinguish between inactive and actively metabolizing filaments after chlorine application to control the bulking of activated sludge. PMID- 16346001 TI - Water-to-Air Transfer and Enrichment of Bacteria in Drops from Bursting Bubbles. AB - An electrostatic induction technique was used to determine both drop size distribution and concentration of bacteria in the film drops produced by bubbles bursting at the surface of a suspension of Serratia marcescens. Film drops are produced from the collapse of the thin film of water that just before bursting separates the air in the bubble from the atmosphere. Bubbles of 1.7-mm diameter produced from 10 to 20 film drops which ranged from <2 mum to over 30 mum in diameter. Half the drops were <10 mum. For bubbles rising a distance of less than 2 cm through the bacterial suspension, bacterial enrichment factors in the drops were between 10 and 20. Electrostatic methods can be used to determine the enrichment of bacteria in film drops as a function of bubble size and distance of rise through the bacterial suspension. PMID- 16346002 TI - Sorption of cadmium by microorganisms in competition with other soil constituents. AB - The fate of cadmium in soil is influenced to a great extent by microbial activity. Microorganisms were compared with abiotic soil components for their ability to sorb Cd from a liquid medium. When the same amount (on a dry weight basis) of bacterial cells (Serratia marcescens and Paracoccus sp.), clay (montmorillonite), or sand was separately incubated in 0.05 M phosphate buffer, pH 7.2, containing 10 ppm of Cd (10 mug/ml), bacterial cells removed the largest quantity of Cd. Dead cells sorbed much more Cd from the medium than live cells. A comparative study of Cd removal from the medium by seven soil bacteria and four fungi did not indicate appreciable differences. With increasing microbial biomass, the relative efficiency of 0.1 M NaOH as an extractant of sorbed Cd increased, whereas the extraction efficiency of 0.005 M DTPA (diethylenetriaminepentaacetic acid) decreased. It appeared that NaOH and DTPA extracted different chemical forms of Cd. This assumption was supported by vastly different correlation coefficients in the relative amount of Cd extracted by the two solvents. PMID- 16346003 TI - Cellular location and some properties of proteolytic enzymes of rumen bacteria. AB - Several physical and chemical techniques were used to extract, and to identify the location of, proteolytic enzymes associated with mixed rumen bacteria. Most activity was removable by gentle physical methods such as shaking and brief blending, without cell disruption, indicating that it was associated with coat and capsular material. Proteases were present also in the cell envelope, corresponding to the inner membrane fraction of gram-negative bacteria, and intracellularly and were removable by detergent and French press treatment. Temperature and pH profiles were obtained for the coat enzymes, likely to be the most important in the digestion of food protein. Inhibition studies indicated that these proteases, and those of the whole bacterial fraction from rumen fluid, were predominantly of the cysteine protease type. PMID- 16346004 TI - Strain-Specific Synthesis of Mycophenolic Acid by Penicillium roqueforti in Blue Veined Cheese. AB - Twenty of 80 strains of Penicillium roqueforti were able to produce up to 600 mg of mycophenolic acid (MPA) liter in 2% yeast extract-5% sucrose broth. Sixty-two of these strains had been isolated from the main blue-veined cheese varieties of western Europe or from starter cultures. Of these 62 dairy strains, only 7 had MPA-producing potential in vitro. These seven strains had all been isolated during the period 1975 to 1981 from the blue cheese of one individual factory. In cheese from the market, MPA (up to 5 mg kg) was only found in samples of this same factory. With MPA-producing and -nonproducing strains for the experimental manufacture of blue cheese, MPA synthesis in cheese was only detected with strains which form MPA in yeast extract-sucrose broth. The maximum MPA level at 4 mg kg was similar to that in commercial cheese. Toxicity of MPA was tested with two established human cell lines (Detroit 98 and Girardi Heart) and one established pig kidney cell line (AmII). PMID- 16346005 TI - Production of 2-Keto-l-Gulonic Acid from d-Glucose by Two-Stage Fermentation. AB - A practical method for the production of calcium 2-keto-l-gulonate (an intermediate in the Reichstein synthesis of l-ascorbic acid) from d-glucose has been established by using a two-stage fermentation system. d-Glucose was first converted to calcium 2,5-diketo-d-gluconate by a mutant strain of Erwinia sp. in a medium containing d-glucose, corn steep liquor, (NH(4))(2)HPO(4), and CaCO(3). After a 26-h cultivation, 328.6 mg of calcium 2,5-diketo-d-gluconate per ml was obtained, with a 94.5% yield from d-glucose. This broth was used directly for the next conversion without removal of cells by treatment with sodium dodecyl sulfate. The stereospecific reduction of calcium 2,5-diketo-d-gluconate to calcium 2-keto-l-gulonate was performed with a mutant strain of Corynebacterium sp. When the cell growth reached a maximum (about 16 h) in a medium containing d glucose, corn steep liquor, NaNO(3), KH(2)PO(4), and trace elements, NaNO(3) was added to the culture, and then the calcium 2,5-diketo-d-gluconate broth was fed over a period of about 50 h. Since the mutant strain requires a hydrogen donor for reduction, the calcium 2,5-diketo-d-gluconate broth was mixed with d-glucose before being fed. The results of four two-stage fermentations in 10-m conventional fermentors showed that an average of 106.3 mg of calcium 2-keto-l gulonate per ml was obtained, with a 84.6% yield from d-glucose, the starting material of calcium 2,5-diketo-d-gluconate production. Calcium 2-keto-l-gulonate was stable in the broth. Neither 2-keto-d-gluconic acid nor 5-keto-d-gluconic acid was detected in the final broth. PMID- 16346006 TI - Effect of sulfide on nitrogen fixation in a stream sediment-water system. AB - Nitrogen fixation (C(2)H(2) reduction) in a sediment-water system was studied under anaerobic incubation conditions. Sodium sulfide at low concentrations stimulated activity, with a twofold increase in C(2)H(4) production occurring in the presence of 8 mumol of S per ml of stream water. Sodium sulfide at concentrations of 16 mumol of S per ml or greater inhibited nitrogen fixation, with 64 mumol of S per ml being completely inhibitory. Sulfide at levels of 16 mumol/ml or above inhibited CO(2) production, and the degree of inhibition increased with increasing concentration of sulfide. Titanium (III) citrate (used to modify Eh levels) stimulated both nitrogen fixation and CO(2) production, but could not duplicate, at any concentration tested, the twofold increase in nitrogen fixation caused by 8 mumol of S per ml. Sulfide additions caused pH changes in the sediment, and when the sediment was adjusted and maintained at pH 7.0 all concentrations of sulfide inhibited nitrogen fixation activity. From considerations of the redox equilibria of H(2), H(2)S, and other sulfur species at various pH values, it appeared that H(2)S was the toxic entity and that HS was less toxic. The observed stimulation of activity was apparently due to a pH change coupled with the concurrent production of HS from H(2)S. PMID- 16346007 TI - Agar-like polysaccharide produced by a pseudomonas species: production and basic properties. AB - A new species of Pseudomonas was isolated that produced copious amounts of an exocellular heteropolysaccharide (PS-60) after incubation for 3 days at 30 degrees C in media containing 3% glucose as a carbon source. The polysaccharide was composed of approximately 46% glucose and 30% rhamnose and, in addition, contained 21% uronic acid and 3% O-acetyl. Upon deacetylation by a mild alkaline treatment, PS-60 produced a brittle, firm, and optically clear gel. This gelling property was thermoreversible. The PS-60 gel exhibited excellent heat stability that withstood autoclaving (i.e., 121 degrees C for 15 min) for several cycles. The gel strength, melting point, and setting point of the polysaccharide were controlled primarily by the concentration of cations. PS-60 was not affected by a variety of enzymes. The results of tests involving various culture media and biochemical test media indicate that PS-60 is an excellent alternative gelling agent to agar. PMID- 16346008 TI - Microbial activities in undecompressed and decompressed deep-seawater samples. AB - Microbial transformations of C-labeled substrates (sodium glutamate, Casamino Acids, glucose, and sodium acetate) were measured in undecompressed seawater samples collected from depths of 1,800 to 6,000 m, during 14- to 21-day incubation periods at in situ temperature (3 degrees C). Each substrate was tested at two concentrations (ca. 0.5 and 5.0 mug/ml) and two in situ pressures. The data were compared to 1-atmosphere (ca. 1.013 x 10 kPa) controls. The rates of C incorporation and CO(2) production as well as the amounts of total substrate utilization were generally lower at pressure than in the decompressed controls but were significantly different for each of the four substrates used. The utilization of acetate was the least affected by pressure; rates were similar to those measured at 1 atmosphere in two out of four experiments. In contrast, transformation rates of the amino acids at pressure averaged to only 38% of those in the controls. A single but reproducible "barophilic" response was observed with glucose as a substrate in samples collected from a depth of 4,500 m at a specific area in the northwestern Atlantic Ocean. Except for this latter set of experiments, the transformation of all substrates showed an increased lag period at pressure as compared to the 1-atmosphere controls. PMID- 16346009 TI - Saccharification of Complex Cellulosic Substrates by the Cellulase System from Clostridium thermocellum. AB - True cellulase activity has been demonstrated in cell-free preparations from the thermophilic anaerobe Clostridium thermocellum. Such activity depends upon the presence of Ca and a thiol-reducing agent of which dithiothreitol is the most promising. Under these conditions, native (cotton) and derived forms of cellulose (Avicel and filter paper) were all extensively solubilized at rates comparable with cellulase from Trichoderma reesei. Maximum activity of the Clostridium cellulase was displayed at 70 degrees C and at pH 5.7 and 6.1 on Avicel and carboxymethylcellulose, respectively. In the absence of substrate at temperatures up to 70 degrees C, carboxymethylcellulase was much more unstable than the Avicel hydrolyzing activity. PMID- 16346010 TI - Rates of mineralization of trace concentrations of aromatic compounds in lake water and sewage samples. AB - The rates of mineralization of phenol, benzoate, benzylamine, p-nitrophenol, and di(2-ethylhexyl) phthalate added to lake water at concentrations ranging from a few picograms to nanograms per milliliter were directly proportional to chemical concentration. The rates were still linear at levels of <1 pg of phenol or p nitrophenol per ml, but it was less than the predicted value at 1.53 pg of 2,4 dichlorophenoxyacetate per ml. Mineralization of 2,4-dichlorophenoxyacetate was not detected in samples of lake water containing 200 ng of the chemical per ml. The slope of a plot of the rate of phenol mineralization in samples of three lakes as a function of its initial concentration was lower at levels of 1 to 100 mug/ml than at higher concentrations. In lake water and sewage supplemented with <60 ng of C-labeled benzoate or phenylacetate per ml, 95 to 99% of the radioactivity disappeared from solution, indicating that the microflora assimilated little or none of the carbon. The extent of mineralization of some compounds in samples of two lakes and sewage was least in the water with the lowest nutrient levels. No mineralization of 2,4-dichlorophenoxyacetate and the phthalate ester was observed in samples of an oligotrophic lake. These data suggest that mineralization of some chemicals at concentrations of <1 mug/ml is the result of activities of organisms different from those functioning at higher concentrations or of organisms that metabolize the chemicals at low concentrations but assimilate little or none of the substrate carbon. PMID- 16346011 TI - Kinetics and extent of mineralization of organic chemicals at trace levels in freshwater and sewage. AB - A sensitive and rapid method was developed to measure the mineralization of C labeled organic compounds at picogram-per-milliliter or lower levels in samples of natural waters and sewage. Mineralization was considered to be equivalent to the loss of radioactivity from solutions. From 93 to 98% of benzoate, benzylamine, aniline, phenol, and 2,4-dichlorophenoxyacetate at one or more concentrations below 300 ng/ml was mineralized in samples of lake waters and sewage, indicating little or no incorporation of carbon into microbial cells. Assimilation of C by the cells mineralizing benzylamine in lake water was not detected. Mineralization in lake waters was linear with time for aniline at 5.7 pg to 500 ng/ml, benzylamine at 310 ng/ml, phenol at 102 fg to 10 mg/ml, 2,4 dichlorophenoxyacetate at 1.5 pg/ml, and di-(2-ethylhexyl) phthalate at 21 pg to 200 ng/ml, but it was exponential at several p-nitrophenol concentrations. The rate of mineralization of 50 and 500 ng of aniline per ml and 200 pg and 2.0 ng of the phthalate per ml increased with time in lake waters. The phthalate and 2,4 dichlorophenoxyacetate were mineralized in samples from a eutrophic but not an oligotrophic lake. Addition to eutrophic lake water of a benzoate-utilizing bacterium did not increase the rate of benzoate mineralization at 34 and 350 pg/ml but did so at 5 and 50 ng/ml. Glucose and phenol reduced the percentage of p-nitrophenol mineralized at p-nitrophenol concentrations of 200 ng/ml but not at 22.6 pg/ml and inhibited the rates of mineralization at both concentrations. These results show that the kinetics of mineralization, the capacity of the aquatic community to assimilate carbon from the substrate or the extent of assimilation, and the sensitivity of the mineralizing populations to organic compounds are different at trace levels than at higher concentrations of organic compounds. PMID- 16346012 TI - Estimations of uronic acids as quantitative measures of extracellular and cell wall polysaccharide polymers from environmental samples. AB - The extracellular polysaccharide polymers can bind microbes to surfaces and can cause physical modification of the microenvironment. Since uronic acids appear to be the components of these extracellular films that are most concentrated in a location outside the cell membrane, a quantitative assay for uronic acids was developed. Polymers containing uronic acids are resistant to quantitative hydrolysis, and the uronic acids, once released, form lactones irreproducibly and are difficult to separate from the neutral sugars. These problems were obviated by the methylation of the uronic acids and their subsequent reduction with sodium borodeuteride to the corresponding alcohol while they were in the polymer and could not form lactones. This caused the polymers to lose the ability to adhere to their substrates, so they could be quantitatively recovered. The hydrolysis of the dideuterated sugars was reproducible and could be performed under conditions that were mild enough that other cellular and extracellular polymers were not affected. The resulting neutral sugars were readily derivatized and then were separated and assayed by glass capillary gas-liquid chromatography. The dideuterated portion of each pentose, hexose, or heptose, identified by combined capillary gas-liquid chromatography and mass spectrometry, accurately provided the proportion of each uronic acid in each carbohydrate of the polymer. Examples of the applications of this methodology include the composition of extracellular polymers in marine bacteria, invertebrate feeding tubes and fecal structures, and the microfouling films formed on titanium and aluminum surfaces exposed to seawater. PMID- 16346013 TI - Effect of interfaces on small, starved marine bacteria. AB - The copiotrophic marine Vibrio sp. strain DW1, shown previously in batch culture to increase in numbers at the onset of starvation and then to form viable small cells with low endogenous respiration, appears to have a survival advantage at interfaces. Vibrio sp. strain DW1 behaved differently at interfaces compared with the aqueous phase under starvation conditions: (i) small cells were observed at an air-water interface without nutrients, (ii) nutrients added to the air-water interface quickly produced larger cells at the surface, (iii) motility persisted many hours longer at the solid-water interface of a dialysis membrane in a microchamber at the onset of starvation, and (iv) regrowth and division at the solid-liquid interface occurred quickly and at nutrient concentrations too low to permit growth in the aqueous phase. It was concluded that, if small starved cells from copiotrophic bacteria can reach an interface, additional survival mechanisms become available to them: (i) interfaces constitute areas of favorable nutrient conditions, and (ii) interfaces lacking a sufficient amount of nutrient, nevertheless, trigger cells to become smaller, thus increasing their surface/volume ratio and the packing density. PMID- 16346014 TI - Pock Formation of Streptomycetes endus with Production of Phage Taillike Particles. AB - Plate (or slope) cultures of endomycin-producing Streptomyces endus (KCC S-0213) showed spontaneously developing pocks which increased in number during subculturing. Neither spore formation nor typical aerial hyphae formation was observed in the pocks, whereas formation of substrate hyphase was not inhibited. Almost all of the hyphae were broken or lysed in the pocks, and many phage tail tiplike particles were observed in the pocks. No self-replication activity was associated with the particles. The particles often formed a hexagonal crystal or a large crystal mass. The production of these particles did not occur in the liquid culture or in young or normal plate cultures having no pocks. These results were similar to those obtained from the plaque-making phenomenon, except for active phage production, in thiostrepton-producing Streptomyces azureus (ATCC 14921), which has been described previously. PMID- 16346015 TI - Effects of Acid on plant litter decomposition in an arctic lake. AB - The effects of acid on the microbial decomposition of the dominant aquatic macrophyte (Carex sp.) in Toolik Lake, Alaska were studied in microcosms during the ice-free season of 1980. Toolik Lake is slightly buffered, deep, and very oligotrophic. Microbial activities, as determined by C-acetate incorporation into extractable lipids, associated with Carex litter were significantly (P < 0.01) reduced within 2 days at pHs of 3.0 and 4.0, but not 5.0, 5.5, or 6.0, as compared with ambient controls (pH 7.4). ATP levels were significantly reduced at pH 3.0, but not at the other pHs tested. After 18 days, microbial activity significantly correlated with weight loss (P < 0.05), nitrogen content (P < 0.01), and C/N ratios (P < 0.01) of the litter, but did not correlate with ATP levels. Scanning electron microscopy of the litter surface revealed that the fungi present at ambient pH did not become dominant at pHs below 5.5, diatoms were absent below pH 4.0, and bacterial numbers and extracellular slime were greatly reduced at pH 4.0 and below. Mineralization of CarexC-lignin-labeled or C cellulose-labeled lignocellulose was reduced at pH 2.0, but not at pH 4.0, 5.0, or 6.0, compared with controls (pH 7). We concluded that if the pH of the water from this slightly buffered lake was sufficiently reduced, rates of litter decomposition would be significantly reduced. PMID- 16346016 TI - Flagella and pili of iron-oxidizing thiobacilli isolated from a uranium mine in northern ontario, Canada. AB - Five strains of Thiobacillus ferrooxidans, which included three recent isolates from a uranium mine, possessed flagella. Three of the strains had several pili per cell. The dimensions, fine structure, and orientation of the flagella were different. Both polar and peritrichous flagella were observed, indicating strain dependent ultrastructural variation in acidophilic thiobacilli. Neither flagella nor pili were detected in eight other strains of T. ferrooxidans and two strains of Thiobacillus acidophilus by electron microscopy, although all of the cultures contained motile cells. PMID- 16346017 TI - Variation in Numbers and Mass of Ciliate Protozoa in the Rumens of Sheep Fed Chaffed Alfalfa (Medicago sativa). AB - Masses and numbers of rumen ciliate protozoa were markedly different in individual sheep fed chaffed alfalfa hay under different feeding regimens. Studies on the ciliate contribution to specific aspects of rumen fermentation should take into account the size of members of each genus in individual animals as well as the numbers present. PMID- 16346018 TI - Purification of Poly-beta-Hydroxybutyrate by Density Gradient Centrifugation in Sodium Bromide. AB - Fractionation of fully sporulated cultures of Bacillus thuringiensis by density gradient centrifugation in NaBr produced two bands which were identified as poly beta-hydroxybutyrate. This technique generated high yields of membrane-bound and unbound granules of exceptional purity and degree of polymerization. PMID- 16346019 TI - Transformation of Streptococcus lactis Protoplasts by Plasmid DNA. AB - Polyethylene glycol-treated protoplasts prepared from Streptococcus lactis LM3302, a lactose-negative (Lac) derivative of S. lactis ML3, were transformed to lactose-fermenting ability by a transductionally shortened plasmid (pLM2103) coding for lactose utilization. PMID- 16346021 TI - Rapid screen for bacteria degrading water-insoluble, solid hydrocarbons on agar plates. AB - [This corrects the article on p. 454 in vol. 43.]. PMID- 16346020 TI - Microbial production of 2,3-butylene glycol from cheese whey. AB - Six microorganisms that produced acetoin or diacetyl or both from glucose were tested for the production of 2,3-butylene glycol from lactose. Bacillus polymyxa and Streptococcus faecalis gave positive results and were tested in unmodified wheys. Cottage cheese whey was unsatisfactory, but B. polymyxa produced large amounts of the glycol in sweet whey, about 60 mmol of glycol per 100 mmol of lactose utilized. Aeration and an increased ratio of surface area to volume of whey enhanced the production of glycol. 2,3-Butylene was separated from the spent whey and from acetoin and diacetyl with a Sephadex G-10 column. PMID- 16346022 TI - Sulfate reduction and methanogenesis in the sediment of a saltmarsh on the East coast of the United kingdom. AB - The rates of sulfate reduction, methanogenesis, and methane loss were measured in saltmarsh sediment at monthly intervals. In addition, dissolved methane and sulfate concentrations together with pS and pH were determined. Methane formation from carbon dioxide, but not from acetate, was detected within the same horizon of sediment where sulfate reduction was most active. Sulfate reduction was about three orders of magnitude greater than annual methanogenesis. The two processes were not separated either spatially or temporally, but occurred within the same layer of sediment at the same time of the year. Their coexistence did not seem to be the result of sulfate-depleted microenvironments within which methanogenesis could occur, but the methanogenic bacteria persisted at very low rates of activity within the same environment as the sulfate reducers. PMID- 16346023 TI - Attached and free-floating bacteria in a diverse selection of water bodies. AB - The contribution of attached and free-floating bacteria to the bacterial numbers and heterotrophic uptake in 44 diverse aquatic environments was studied. A factor analysis reduced the variability of the raw data base to three major factors explaining 53.6% of total variance. These factors were (i) salinity, (ii) heterotrophic uptake, and (iii) particulate load. A cluster analysis categorized the 44 habitats into five distinct environmental types based on these three factors. There was no significant pattern in the distribution of attached versus free-floating bacteria when assessed by epifluorescent microscopy. However the contribution of attached bacteria to the uptake of an amino acids mix was reduced in marine waters. Heavy particulate loads resulted in an increased percentage uptake of amino acids and glucose from the attached bacteria. Uptake response was found to be substrate specific especially in oliogotrophic freshwater. Amino acid uptake was more associated with the attached fraction, whereas glucose uptake was mediated more by the free-floating fraction. PMID- 16346024 TI - Nitrogenase Activity Associated with Halodule wrightii Roots. AB - Nitrogen fixation (acetylene reduction) associated with roots of the seagrass Halodule wrightii was measured offshore near Beaufort and Moorhead City, N.C. Rates of acetylene reduction were higher in aerobic than in anaerobic assays and were linear for up to 5 days. The temperature range for acetylene reduction was 15 to 35 degrees C with a maximum activity at 35 degrees C. Nitrogenase activity was shown to vary seasonally with highest activities occurring during warmer summer months (23 mug of N(2) fixed per m per day). At in situ temperature, nitrogenase activities associated with surface-sterilized and non-surface sterilized roots were similar. One morphological bacterial type was isolated from surface-sterilized roots and identified as Klebsiella pneumoniae type 4B. PMID- 16346025 TI - Improved method for determination of respiring individual microorganisms in natural waters. AB - A method is reported that combines the microscopic determinations of specific, individual, respiring microorganisms by the detection of electron transport system activity and the total number of organisms of an estuarine population by epifluorescence microscopy. An active cellular electron transport system specifically reduces 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) to INT-formazan, which is recognized as opaque intracellular deposits in microorganisms stained with acridine orange. In a comparison of previously described sample preparation techniques, a loss of >70% of the counts of INT-reducing microorganisms was shown to be due to the dissolution of INT formazan deposits by immersion oil (used in microscopy). In addition, significantly fewer fluorescing microorganisms and INT-formazan deposits, both 80% of total sulfur) in sediment. Although sediment sulfur is predominantly organic, sulfate reduction is the most significant process in terms of the quantities of sulfur transformed in surface sediments. Rates of sulfate reduction in these sediments average 7 mmol/m per day. On an annual basis, this rate is 19-fold greater than net rates of organic sulfur mineralization and 65-fold greater than sulfate ester hydrolysis. PMID- 16346038 TI - Solvent Production and Morphological Changes in Clostridium acetobutylicum. AB - The morphological and cytological changes which occurred in Clostridium acetobutylicum P262 during the production of acetone, butanol, and ethanol in an industrial fermentation medium were identified and correlated with the growth and physiological changes. The swollen, cigar-shaped clostridial forms were involved in the conversion of acids to neutral solvents, and there was a correlation between the number of clostridial forms and the production of solvents. Sporulation mutants which were unable to form clostridial stages (cls mutants) did not produce solvents. Oligosporogenous mutants which showed reduced clostridial stage formation produced intermediate levels of solvents. Sporulation mutants blocked after the clostridial stage, which were unable to form mature spores (spo mutants), produced normal levels of solvents. PMID- 16346039 TI - Isolation and properties of ferromanganese-depositing budding bacteria from baltic sea ferromanganese concretions. AB - Hyphal budding bacteria were observed by electron microscopy in thin sections of surface material from Baltic Sea ferromanganese concretions. Similar bacteria were also observed in and isolated from enrichment cultures prepared from the same concretion material. Three morphologically similar strains of Mn-Fe depositing budding bacteria were isolated from the enrichment cultures. Strain B 4 possessed extracellular anionic polymers that accumulated Mn oxides. Mn deposition by B-4 was inhibited by elevated concentrations of Mn, 0.05% glutaraldehyde, 0.1 mM HgCl(2), and heating at 93 degrees C for 15 min, suggesting the participation of an enzyme protein in the Mn-depositing activity. PMID- 16346040 TI - Formation of Crystalline delta-Endotoxin or Poly-beta-Hydroxybutyric Acid Granules by Asporogenous Mutants of Bacillus thuringiensis. AB - Parental strains and asporogenous mutants of Bacillus thuringiensis subspp. kurstaki and aizawai produced high yields of delta-endotoxin on M medium, which contained 330 mug of potassium per ml, but not on ST and ST-a media, each of which contained only 11 mug of potassium per ml. On ST and ST-a media, refractile granules were formed instead. These granules had no insecticidal activity against silkworms and were isolated and identified as poly-beta-hydroxybutyric acid. Supplementation of the potassium-deficient ST-a medium with 0.1% KH(2)PO(4) (3.7 mM) led to the formation of crystalline delta-endotoxin. The replacement of KH(2)PO(4) with equimolar amounts of KCl, KNO(3), and potassium acetate or an equivalent amount of K(2)SO(4) had a similar effect, whereas the addition of an equimolar amount of NaH(2)PO(4) or NH(4)H(2)PO(4) did not cause the endotoxin to form. An asporogenous mutant, B. thuringiensis subsp. kurstaki strain 290-1, produced delta-endotoxin on ST-a medium supplemented with 3 mM or more potassium but formed only poly-beta-hydroxybutyric acid granules on the media containing 4)-linked glucan. These products correlate with the known structure of the gum. The complexity of the product mixture indicated that the xanthanase was a mixture of carbohydrases. The xanthanase complexes were similar to one another in temperature stability, pH and temperature optima, degree of substrate degradation, and enzymolysis products. Differences in pH stability, salt tolerance, recoverability, and yields of enzyme were observed. PMID- 16346069 TI - Phenylpropanoic Acid: Growth Factor for Ruminococcus albus. AB - Phenylpropanoic acid accounted for part of the stimulatory effect of rumen fluid on the rate of growth and of cellulose digestion by cultures of Ruminococcus albus strain 8 grown on a chemically defined medium. As little as 3 muM concentration gave maximum response. PMID- 16346071 TI - Root Surface Association in Relation to Nodulation of Medicago sativa. AB - Nine strains of Rhizobium meliloti, ranging in competitive ability on Medicago sativa from excellent to poor in autoclaved soils, were paired in 29 combinations and used to inoculate M. sativa in a liquid rooting medium. A positive correlation (r = 0.545) between strain ratios in nodules after 28 days and root surface cell ratios after 7 days was determined. Two cell fractions from the root surface, representing loosely and firmly adhering cells, were investigated. Infectivity was linked to the more firmly adhering cells. A significant relationship was established between the cell ratios of competing strains in the two fractions. In another experiment, adherence of cells of both infective and noninfective Rhizobium strains to roots of M. sativa and Trifolium repens was demonstrated; the ratios of loosely to firmly adhering cells on the root surface were significantly narrower with the infective combinations than with noninfective strain-legume associations. PMID- 16346070 TI - Synergistic Interaction Between Anabaena and Zoogloea spp. in Carbon Dioxide Limited Continuous Cultures. AB - Flocs consisting of Anabaena and Zoogloea spp. were used as a model system for the study of planktonic phototroph-heterotroph interactions. In CO(2)-limited continuous culture (3.2 mumol of NaHCO(3) liter h, 1.5 mumol of glucose liter h, pH 8.5, D = 0.026 h), the biomass of the phototroph increased 8.6-fold due to association. However, direct CO(2) exchange accounted for only a 3.8-fold increase. When the glucose supply rate was increased to 7.5 mumol liter h, there was a 26-fold increase in biomass. When CO(2) was supplied in excess, there was no difference due to association. In batch culture, using the same medium, the specific growth rate was 0.029 h for the phototroph alone and 0.047 h for the phototroph in association with the heterotroph. The stimulatory effect of the heterotroph was found only under CO(2)-limiting conditions and was directly related to the concentration of organic matter supplied in the medium. Both the biomass and the growth rate of the Anabaena sp. were increased by association with the Zoogloea sp. Thus, dissolved organic matter may substitute for CO(2) to maximize both growth rate and biomass production by phototrophs when heterotrophic bacteria are present. PMID- 16346072 TI - Competitive Abilities of Rhizobium meliloti Strains Considered to Have Potential as Inoculants. AB - Twenty four strains of Rhizobium meliloti considered to have potential for inoculant production were grouped in pairs and tested for their ability to compete for nodulation on Medicago sativa, Medicago truncatula, and Medicago littoralis. At the outset, each pair of strains, which consisted of a wild type and a selected streptomycin-resistant mutant of another strain, was tested in an autoclaved soil. Six strain pairs, each consisting of a good and a poor competitor, reacted consistently when tested in each of five other autoclaved soils; eight pairs consisting of strains with comparable competitive abilities varied in their reactions in some of the soils, or even in the same soil when retested. An effect of soil pH on competitive ability was observed with some of these strains. Not all of the strains identified as good competitors on one or more of the Medicago spp. in the autoclaved soils were able to nodulate these plants satisfactorily in a field soil containing an established population of R. meliloti. Strain RF24, which seemed to be the best competitor on each of the three Medicago spp., grouped among the less effective strains on two of the legumes. Two strains of R. meliloti frequently used for inoculant production differed markedly with regard to competitive ability; this places some doubt on the relevancy of singling out competitive ability for special attention when selecting a strain for inoculant production. PMID- 16346073 TI - Mass spectrometric quantification of indole-3-acetic Acid in Rhizobium culture supernatants: relation to root hair curling and nodule initiation. AB - Indole-3-acetic acid (IAA) has been unequivocally identified in culture supernatants of Rhizobium strains by gas chromatography-mass spectrometry. A method for accurately quantitating IAA in bacterial culture supernatants, employing deuterium-labeled IAA as an internal standard, has been developed. Similar IAA concentrations were found in culture supernatants of chosen Rhizobium mutants (defective in nodule formation) and their corresponding parent strains. Since some of the mutants are known to adhere to root hairs, it can be concluded that root hair curling is not simply a consequence of IAA production by rhizobia. PMID- 16346074 TI - Role of aerobic microbial populations in cellulose digestion by desert millipedes. AB - I examined the role of aerobic microbial populations in cellulose digestion by two sympatric species of desert millipedes, Orthoporus ornatus and Comanchelus sp. High numbers of bacteria able to grow on media containing cellulose, carboxymethyl cellulose, or cellobiose as the substrate were found in the alimentary tracts of the millipedes. Enzyme assays indicated that most cellulose and hemicellulose degradation occurred in the midgut, whereas the hindgut was an important site for pectin degradation. Hemicellulase and beta-glucosidase in both species and possibly C(x)-cellulase and pectinase in O. ornatus were of possible microbial origin. Degradation of [C]cellulose by millipedes whose gut floras were reduced by antibiotic treatment and starvation demonstrated a reduction in CO(2) release and C assimilation and an increase in C excretion over values for controls. It appears that the millipede-bacterium association is mutualistic and makes available to millipedes an otherwise mostly unutilizable substrate. Such an association may be an important pathway for decomposition in desert ecosystems. PMID- 16346075 TI - Dilution of liquid Rhizobium cultures to increase production capacity of inoculant plants. AB - Experiments were undertaken to test whether peat-based legume seed inoculants, which are prepared with liquid cultures that have been deliberately diluted, can attain and sustain acceptable numbers of viable rhizobia. Liquid cultures of Rhizobium japonicum and Rhizobium phaseoli were diluted to give 10, 10, or 10 cells per ml, using either deionized water, quarter-strength yeast-mannitol broth, yeast-sucrose broth, or yeast-water. The variously diluted cultures were incorporated into gamma-irradiated peat, and the numbers of viable rhizobia were determined at intervals. In all of the inoculant formulations, the numbers of rhizobia reached similarly high ceiling values by 1 week after incorporation, irrespective not only of the number of cells added initially but also of the nature of the diluent. During week 1 of growth, similar multiplication patterns of the diluted liquid cultures were observed in two different peats. Numbers of rhizobia surviving in the various inoculant formulations were not markedly different after 6 months of storage at 28 degrees C. The method of inoculant preparation did not affect the nitrogen fixation effectiveness of the Rhizobium strains. PMID- 16346076 TI - Single-cell protein production in alkaline mesquite wood extracts. AB - A nine-member, mixed, cellulolytic, bacterial culture was used to evaluate the effects of sodium hydroxide normality, length and temperature of treatment, and the ratio of volume of alkali to mesquite wood on the suitability of alkali mesquite wood extractives as nutrients for bacterial growth. The presence or absence of air during the extraction process did not significantly affect results. The amount of lignin extracted and the total loss in weight of the wood during extraction were correlated to both alkali concentration and temperature. Neutralized extracts supported bacterial growth; growth was inversely related to the final salt concentration of the neutralized extract. Deionized extracts were superior to acid-neutralized extracts for the support of bacterial growth. The optimum conditions for extraction were 2.5 N NaOH at 30 degrees C for 12 h. The study demonstrated that nutrients as well as growth inhibitory compounds are released from wood by alkali treatment. This study of alkali wood extracts and the previous study of washed alkali treated wood residues provide a data base for the optimization of alkali treatments of hard woods that are to be used as nutrients for the growth of cellulolytic cultures. PMID- 16346077 TI - Inhibition of Expression of Fusarium semitectum and Other Fungi in Immature Cottonseed. AB - Fusarium semitectum Berkeley and Ravenel and other nonpathogenic fungi appear to contaminate immature cottonseed but cannot express themselves until immediately after seed maturation. It is hypothesized that catechin prevents fungal outgrowth and that inhibition is released when catechin is polymerized during seed maturation. PMID- 16346078 TI - Field observations of methane concentrations and oxidation rates in the southeastern bering sea. AB - Measurements of methane oxidation rates were made in southeastern Bering Sea water samples with [C]methane. The rate at which CO(2) evolved from samples exposed to one methane concentration was defined as the relative methane oxidation rate. Rate determinations at three methane concentrations were used to estimate methane oxidation kinetics. The rate constant calculated from the kinetics and the observed methane concentration in the same water sample were used to calculate an in situ methane oxidation rate and the turnover time. First order kinetics were observed in essentially all experiments in which methane oxidation kinetics were measured. Relative methane oxidation rates were greater in waters collected at inshore stations than at the offshore stations and were greater in bottom samples than in surface samples. In most water samples analyzed, there was essentially no radioactivity associated with the cells. The resulting respiration percentages were therefore very high with a mean of >98%. These data suggest that most of the methane was used by the microflora as an energy source and that very little of it was used in biosynthesis. The relative methane oxidation rates were not closely correlated with methane concentrations and did not appear to be linked to either oxygen or dissolved inorganic nitrogen concentrations. However, there was a significant correlation with relative microbial activity. Our data suggest that the methane oxidizers were associated with the general microbial heterotrophic community. Since these organisms did not appear to be using methane as a carbon source, it is unlikely that they have been isolated and identified as methane oxidizers in the past. PMID- 16346079 TI - Isolation of frankia strains from alder actinorhizal root nodules. AB - A simple procedure, based on the rapid filtration and washing of Frankia vesicle clusters, was devised for the isolation of Frankia strains from alder actinorhizal root nodules. Of 46 Alnus incana subsp. rugosa nodules prepared, 42 yielded isolates. A simple medium containing mineral salts, Casamino Acids, and sodium pyruvate proved to be the most effective for isolation. In general, colonies appeared 6 to 20 days after inoculation. On the basis of hyphal morphology, two distinct types of Frankia strains were characterized. Randomly selected isolates were tested for infectivity, and all formed root nodules on A. glutinosa. Because of its simplicity and efficiency, the procedure is an improved method for the study of Frankia diversity in alder root nodules. PMID- 16346080 TI - Effects of surface area and flow rate on marine bacterial growth in activated carbon columns. AB - The colonization of granular activated carbon columns by bacteria can have both beneficial and potentially detrimental consequences. Bacterial growth on the carbon surface can remove adsorbed organics and thus partially regenerate the carbon bed. However, growth can also increase the levels of bacteria in the column effluents, which can adversely affect downstream uses of the treated water. This study of a sand column and several activated carbon columns demonstrated that considerable marine bacterial growth occurred in both sand and carbon columns and that this growth increased the number of bacteria in column effluents. Activated carbon supported approximately 50% more bacteria than did sand. Bacterial growth on activated carbon was reduced by increasing the flow rate through a carbon column and increasing the carbon particle size. Scanning electron micrographs showed that bacteria preferred to attach in the protected crevices on both the sand and carbon surface. The results of this study indicated that the colonization of activated carbon by marine bacteria was enhanced because of carbon's high surface area, its rough surface texture, and its ability to absorb organic materials. PMID- 16346081 TI - Alteration of the Trifoliin A-Binding Capsule of Rhizobium trifolii 0403 by Enzymes Released from Clover Roots. AB - The effect of white clover root exudate on capsules of Rhizobium trifolii 0403 was examined. The clover lectin trifoliin A was detected in root exudate of two clover varieties by indirect immunofluorescence with antibody against this lectin purified from clover seed. Trifoliin A bound uniformly to encapsulated, heat fixed cells during 1 h of incubation with root exudate. After 4 to 8 h of incubation, trifoliin A was only bound to one pole of the cells. Transmission electron microscopy showed that the capsule itself was altered. The disorganization of the acidic polymers of the capsule began in the equatorial center of the rod-shaped cell and then progressed toward the poles at unequal rates. Trifoliin A could no longer be detected on heat-fixed cells after 12 h of incubation with root exudate. However, trifoliin A was detected in situ on one pole of cells grown for 4 days in the clover root environment of Fahraeus slide cultures. Inhibition studies with the hapten 2-deoxy-d-glucose showed that trifoliin A in root exudate had a higher affinity for one of the cell poles. Immunoelectrophoresis was used to monitor the alteration of the extracellular polysaccharides from R. trifolii 0403 by concentrated root exudate. These polysaccharides were converted into products which eventually lost their ability to immunoprecipitate with homologous antibody. This progressive loss of antigenic reactivity proceeded more rapidly with root exudate from seedlings grown under nitrogen-free conditions than with root exudate from plants grown with 15 mM KNO(3). The root exudate, depleted of trifoliin A by immunoaffinity chromatography, was still able to alter the capsule of R. trifolii 0403. Reconstitution experiments showed that the substance(s) in root exudate which induced this alteration of the capsule was of a high molecular weight, heat labile, trypsin sensitive, and antigenically unrelated to trifoliin A. A variety of glycosidase activities were also detected in the fraction depleted of trifoliin A. These results suggest that enzymes in clover root exudate alter the trifoliin A-binding capsule in a way which would favor polar attachment of R. trifolii to clover root hairs. PMID- 16346082 TI - Microbial metal tolerance in bermuda carbonate sediments. AB - The recovery of aerobic heterotrophic bacteria from Bermuda carbonate sediments on metal-supplemented media varied as much as 44-fold over a 15-cm depth. Distributional relationships with sulfate-reducing bacteria and sediment character indicated that metal tolerance was a function of metal bioavailability. PMID- 16346083 TI - Use of gentamicin in axenic culturing of ectomycorrhizal plants. AB - Gentamicin is inhibitory to bacteria associated with Pinus ectomycorrhizae, but does not appear to affect the growth and metabolic activity of the fungus or root. PMID- 16346085 TI - Estimations of uronic acids as quantitative measures of extracellular and cell wall polysaccharide polymers from environmental samples. AB - [This corrects the article on p. 1153 in vol. 43.]. PMID- 16346084 TI - Widespread distribution of ability to oxidize manganese among freshwater bacteria. AB - Manganese-oxidizing heterotrophic bacteria were found to comprise a significant proportion of the bacterial community of Lake Washington (Seattle, Wash.) and Lake Virginia (Winter Park, Fla.). Identification of these freshwater bacteria showed that members of a variety of genera are capable of oxidizing manganese. Isolates maintained in the laboratory spontaneously lost the ability to oxidize manganese. A direct correlation was found between the presence of plasmid DNA and the ability of the organism to oxidize manganese. PMID- 16346086 TI - Airstream fractionation of vesicular-arbuscular mycorrhizal fungi: concentration and enumeration of propagules. AB - Spores and fragments of vesicular-arbuscular mycorrhizal fungi in dry soils were concentrated up to 100-fold when the soils were partitioned by fluidization and elutriation with a series of upward airstreams at progressively increasing velocities. The propagules were transported with the finer soil particles according to their equivalent spherical diameters. The system was used to predict the transport of propagules by wind. Concentrated propagules were rapidly separated from the soil particles in each soil fraction by an aqueous flotation method. The technique is proposed as a quantitative method for estimating the numbers of spores and fragments of mycorrhizae. The scheme includes a viability test that was used to differentiate between potentially infective propagules and those that were either dormant or incapable of regrowth. PMID- 16346087 TI - Kinetics of Formate Metabolism in Methanobacterium formicicum and Methanospirillum hungatei. AB - The kinetics of formate metabolism in Methanobacterium formicicum and Methanospirillum hungatei were studied with log-phase formate-grown cultures. The progress of formate degradation was followed by the formyltetrahydrofolate synthetase assay for formate and fitted to the integrated form of the Michaelis Menten equation. The K(m) and V(max) values for Methanobacterium formicicum were 0.58 mM formate and 0.037 mol of formate h g (dry weight), respectively. The lowest concentration of formate metabolized by Methanobacterium formicicum was 26 muM. The K(m) and V(max) values for Methanospirillum hungatei were 0.22 mM and 0.044 mol of formate h g (dry weight), respectively. The lowest concentration of formate metabolized by Methanospirillum hungatei was 15 muM. The apparent K(m) for formate by formate dehydrogenase in cell-free extracts of Methanospirillum hungatei was 0.11 mM. The K(m) for H(2) uptake by cultures of Methanobacterium formicicum was 6 muM dissolved H(2). Formate and H(2) were equivalent electron donors for methanogenesis when both substrates were above saturation; however, H(2) uptake was severely depressed when formate was above saturation and the dissolved H(2) was below 6 muM. Formate-grown cultures of Methanobacterium formicicum that were substrate limited for 57 h showed an immediate increase in growth and methanogenesis when formate was added to above saturation. PMID- 16346088 TI - Metabolism of the Plasticizer di-n-Butylphthalate by Pseudomonas pseudoalcaligenes Under Anaerobic Conditions, with Nitrate as the Only Electron Acceptor. AB - The metabolism of di-n-butylphthalate by a denitrifying strain of Pseudomonas pseudoalcaligenes B20b1 was studied under anaerobic conditions, with nitrate as the only electron acceptor. Thin-layer chromatography and mass spectral analysis of culture extracts (20 days at 30 degrees C) showed mono-n-butylphthalate and phthalic acid as the only products, suggesting that one butanol moiety served essentially as the carbon source for growth and denitrification. N(2) and N(2)O were detected by gas chromatography. In contrast to aerobic metabolism, phthalate was not degraded further if nitrate was the only electron acceptor. PMID- 16346089 TI - Quantitative study of nodulation competitiveness in Rhizobium strains. AB - We compared the nodulation competitiveness of three strains of Rhizobium leguminosarum by counting the number of nodules formed on faba bean plants after the application at sowing time of different concentrations of the strains to soils already containing Rhizobium strains of the same species. A relationship of type y = ax was found to exist between the ratio of the nodules formed by the applied inoculum strain to the nodules formed by the soil strains and the ratio of Rhizobium cells in the inoculum to the cells in the soil. This relationship was also confirmed in another competition experiment in which two R. meliloti strains of identical competitiveness were mixed in various proportions. The relationship can also be applied to the majority of results reported in the literature. Should it prove to be more widely applicable, it could be used to estimate the relative competitiveness of Rhizobium strains and thus predict the performance of an inoculum in a given soil. PMID- 16346090 TI - Influence of plant phenolic acids on growth and cellulolytic activity of rumen bacteria. AB - Isolated rumen bacteria were examined for growth and, where appropriate, for their ability to degrade cellulose in the presence of the hydroxycinnamic acids trans-p-coumaric acid and trans-ferulic acid and the hydroxybenzoic acids vanillic acid and 4-hydroxybenzoic acid. Ferulic and p-coumaric acids proved to be the most toxic of the acids examined and suppressed the growth of the cellulolytic strains Ruminococcus albus, Ruminococcus flavefaciens, and Bacteroides succinogenes when included in a simple sugars medium at concentrations of >5 mM. The extent of cellulose digestion by R. flavefaciens and B. succinogenes but not R. albus was also substantially reduced. Examination of rumen fluid from sheep maintained on dried grass containing 0.51% phenolic acids showed the presence of phloretic acid (0.1 mM) and 3-methoxyphloretic acid (trace) produced by hydrogenation of the 2-propenoic side chain of p-coumaric and ferulic acids, respectively. The parent acids were found in trace amounts only, although they represented the major phenolic acids ingested. Phloretic and 3 methoxyphloretic acids proved to be considerably less toxic than their parent acids. All of the cellulolytic strains (and Streptococcus bovis) showed at least a limited ability to hydrogenate hydroxycinnamic acids, with Ruminococcus spp. proving the most effective. No further modification of hydroxycinnamic acids was produced by the single strains of bacteria examined. However, a considerable shortfall in the recovery of added phenolic acids was noted in media inoculated with rumen fluid. It is suggested that hydrogenation may serve to protect cellulolytic strains from hydroxycinnamic acids. PMID- 16346091 TI - Cowpea rhizobia producing dark nodules: use in competition studies. AB - During a program of screening rhizobia from West Africa, it was found that some strains produced nodules of unusually dark appearance on cowpeas, but not on peanuts, soybeans, pigeon peas, or mung beans. The dark pigmentation was in the bacteroid zone, was not correlated with nodule effectiveness, and was additional to the leghemoglobin pigment. Only rhizobial strains with a nongummy ("dry") colony morphology produced dark nodules. Visually distinguishable pink and dark nodules formed on the same root when a mixture of pink and dark strains was applied as inoculum. The dark-nodule phenotype was therefore appraised as a marker and found to be useful for studying nodulation competition with strains of the orthodox pink-nodule type. The competitiveness of 10 pink-nodule strains was examined relative to a black-nodule strain, IRc 256; a range of competitiveness was obtained of less competitive than, equally competitive to, or more competitive than IRc 256. Patterns of primary (early) nodulation were generally the same as patterns of secondary (later) nodulation. Mixed infections by dark and pink strains produced piebald nodules, the frequency of occurrence of which was much greater among primary than among secondary nodules. PMID- 16346092 TI - Saccharomyces cerevisiae Mutants Resistant to Catabolite Repression: Use in Cheese Whey Hydrolysate Fermentation. AB - Mutants of an industrial-type strain of Saccharomyces cerevisiae which rapidly and completely fermented equimolar mixtures of glucose and galactose to ethanol were isolated. These mutants fell into two general phenotypic classes based upon their fermentation kinetics and enzyme induction patterns. One class apparently specifically effects the utilization of galactose and allows sequential utilization of first glucose and then galactose in an anaerobic fermentation. The second class of mutants was resistant to general catabolite repression and produced maltase, invertase, and galactokinase in the presence of repressive levels of glucose. These mutants were completely dominant and appear to represent an as yet undescribed class of mutant. PMID- 16346093 TI - Variations in Ability of Rhizobium japonicum Strains To Nodulate Soybeans and Maintain Fixation in the Presence of Nitrate. AB - This study investigated differences in sensitivity to nitrate of soybean (Glycine max cv. Davis) symbioses with 16 different Rhizobium japonicum strains. When nitrate (20 mM) was added to established symbioses, there were no significant differences in the degree of inhibition of acetylene reduction for any of the 16 strains. When nitrate was present during the establishment of nodules, high levels of nitrate (10 mM) were equally inhibitory on all symbioses, whereas specific strain effects appeared at low (0.5 mM) to medium (2.0 mM) levels of nitrate. At 1.5 mM nitrate in solution culture, the days to emergence of nodules varied from less than 10 (CB:1809, Nit61A118) to more than 16 (11 of 16 strains). In a clay-pot trial maintained at the low nitrate level (0.5 mM), symbioses with CB:1809 increased total nodule mass by 30% relative to nitrate-free controls. In the presence of 2.0 mM nitrate, CB:1809 maintained total nodule mass. For the remaining 6 strains tested, total nodule mass decreased to below the levels of the nitrate-free controls. In a separate clay pot trial, CB:1809 increased its competitive ability relative to USDA:110 when nitrate was added. If no nitrate was added, CB:1809 occupied 0.97 times as many nodules as USDA:110; when 10 mM nitrate was added, CB:1809 occupied 1.75 times as many nodules as USDA:110. Attempts to select nitrogen-adapted substrains of R. japonicum through sequential isolation and infection of plants grown on nitrate were not successful. PMID- 16346094 TI - Competitive Dominance by a Bacteriocin-Producing Vibrio harveyi Strain. AB - Vibrio (Beneckea) harveyi, a bioluminescent marine bacterium, has been shown to produce a bacteriocin-like substance the production of which is mediated by a plasmid. This substance is assumed to be proteinaceous because of its sensitivity to certain proteolytic enzymes. It is stable at low temperatures and can be concentrated by ammonium sulfate precipitation or negative-pressure dialysis. The molecular weight of the bacteriocin was determined to be 2.4 x 10 by molecular exclusion chromatography. Competition experiments indicated that bacteriocin producing strains predominated over cured variants of the same strain in broth culture experiments. We studied several environmental parameters (pH, salinity, temperature, nutrient concentration) to determine their effects on the competitive advantage bestowed on a bacteriocin-producing strain. Under simulated free-living conditions, no competitive advantage attributable to bacteriocin production was observed. In a simulated enteric habitat, a bacteriocin-producing strain showed dramatic (>90%) inhibition of the sensitive strain within 24 h. PMID- 16346095 TI - Effect of sorption on mineralization of low concentrations of aromatic compounds in lake water samples. AB - Montmorillonite-benzylamine complexes were formed immediately upon addition of 20 pg to 20 mug of amine per ml of suspensions containing the clay. The extent of amine sorbed was a linear function of equilibrium amine concentration in lake water. Increases in the clay concentration decreased the percentage of the organic compound that was mineralized at amine levels of 20 pg to 200 ng, but not at 20 mug/ml. A larger percentage of the chemical was released from the complex during mineralization in the presence of high clay concentrations than in the presence of low clay concentrations. The rates of desorption and mineralization increased linearly with benzylamine levels up to 200 ng/ml. Montmorillonite did not enhance mineralization rates at amine levels of 200 ng/ml or lower, but it was stimulatory at 20 mug/ml. Except at high amine and clay concentrations, mineralization was more rapid than desorption during the early periods of decomposition when the amine concentration in solution was relatively high. However, relative to the microbial demand, desorption was more rapid during later periods of decomposition when the amine level in solution was very low. Mineralization of benzoate was not usually affected by montmorillonite, kaolinite, or glass beads. More than 90% of the carbon from benzylamine and benzoate was often mineralized when the substrate concentration was 250 ng/ml or less. After incubation of the chemical in lake water, none of the radioactivity from benzylamine was in the particulate fraction containing natural sediment and microbial cells. The data indicate that clay may have a significant effect on the microbial decomposition of low concentrations of certain organic compounds. PMID- 16346096 TI - Biomagnification of aroclor 1242 in decomposing spartina litter. AB - The accumulation of a polychlorinated biphenyl (PCB) mixture (Aroclor 1242) in the process of detritus formation by a shredded marshgrass (Spartina alterniflora) under aerobic conditions was monitored in percolators for 4 months at 20 degrees C. Dissolved PCB in the influent solution was 14 to 16 mug/liter. Parameters monitored in addition to PCB accumulation were CO(2) evolution, NH(4) uptake, NO(3) production, and total organic nitrogen and carbon in the effluent. An NaN(3-) poisoned control served to assess nonbiological PCB absorption. Up to 90% of the PCB dissolved in the influent water was removed by the biologically active detritus. Biomagnification resulted in three to four times higher PCB concentrations in the active detritus than in the abiotic control. Evolution of CO(2) was slightly depressed by PCB, but the overall quality (C:N ratio) of the detritus was not affected. The results indicate that bioaccumulation of PCB in detritus is an important means of entry for this pollutant into estuarine food webs. PMID- 16346097 TI - Lognormal distribution of epiphytic bacterial populations on leaf surfaces. AB - Total populations of epiphytic bacteria and selected components thereof were determined on sets of 24 to 36 individual leaves (corn, rye) or leaflets (snap bean, soybean, tomato) of field-grown plants by washing and dilution plating. In general, levels of component populations (e.g., bacteria that are ice nucleation active) were quantitatively more variable from leaf to leaf within a set than were total epiphytic bacterial populations. Populations of a given component frequently varied by a factor of 100 to 1,000 within a set of leaves. Total bacterial populations usually varied by a factor of about 10. For each set of leaves, total and component epiphytic bacterial populations were found to approximate a lognormal distribution by the Shapiro-Wilk test for normality. Due to the lognormal distribution of epiphytic bacterial populations, estimates of population size based on the common practice of using bulked samples (wherein several leaves are washed together) will overestimate the population median by a factor of approximately 1.15sigma. From the known probability distribution of bacterial populations, the frequency with which a given bacterial population size is met or exceeded on individual leaves can be estimated. If the bacterial component is phytopathogenic, the frequency estimates could be used to relate quantitatively pathogen populations and disease incidence. PMID- 16346098 TI - Commensalistic Interaction Between Lactobacillus acidophilus and Propionibacterium shermanii. AB - Propionibacterium shermanii and Lactobacillus acidophilus were grown in batch mixed culture in a 5-liter fermenter under controlled conditions of pH 5.8 and 35 degrees C on a semisynthetic medium with glucose as an energy source. Cellular efficiencies and fermentation balances were developed for this pair and compared with P. shermanii grown in pure culture on glucose, lactate, and a mixture of these substrates and with L. acidophilus grown on glucose. P. shermanii had ATP yield coefficient values of 17 for each substrate alone but had an average value of 30 for substrate mixtures. Growth rates were similar for P. shermanii on glucose or lactate but higher cell yields were observed for glucose. P. shermanii used both lactate and glucose in mixed substrate until lactate was exhausted, and growth rates slowed thereafter. L. acidophilus had a similar ATP yield coefficient of 15 but produced lower cell yields than did P. shermanii on glucose. Mixed culture of both microorganisms on glucose resulted in much faster and nearly equal growth rates for both and no lactate accumulation in the medium. Acetic acid production rates per generation were lower in mixed culture, suggesting use by the growing culture. The cause of the synergistic effect was not determined but may be due to the rapid production and removal of lactate or CO(2) enhancement in mixed culture. PMID- 16346099 TI - Ultrastructural Localization of Hydrogen Peroxide Production in Ligninolytic Phanerochaete chrysosporium Cells. AB - Previous studies have shown that the hydroxyl radical derived from hydrogen peroxide (H(2)O(2)) is involved in lignin degradation by Phanerochaete chrysosporium. In the present study, the ultrastructural sites of H(2)O(2) production in ligninolytic cells of P. chrysosporium were demonstrated by cytochemically staining cells with 3,3'-diaminobenzidine (DAB). Hydrogen peroxide production, as evidenced by the presence of oxidized DAB deposits, appeared to be localized in the periplasmic space of cells from ligninolytic cultures grown for 14 days in nitrogen-limited medium. When identical cells were treated with DAB in the presence of aminotriazole, periplasmic deposits of oxidized DAB were not observed, suggesting that the deposits resulted from the H(2)O(2)-dependent peroxidatic oxidation of DAB by catalase. Cells from cultures grown for 3 or 6 days in nitrogen-limited medium or for 14 days in nitrogen-sufficient medium had little ligninolytic activity and low specific activity for H(2)O(2) production and did not contain periplasmic oxidized DAB deposits. The results suggest that in cultures grown in nitrogen-limited medium, there is a positive correlation between the occurrence of oxidized DAB deposits, the specific activity for H(2)O(2) production in cell extracts, and ligninolytic activity. PMID- 16346100 TI - Aerobic, Selenium-Utilizing Bacillus Isolated from Seeds of Astragalus crotalariae. AB - Bacillus sp. strain SS, an aerobic, gram-positive sporeformer, was isolated from seeds of Astragalus crotalariae, a selenium-accumulating plant. This bacillus grew in a nutrient broth (containing beef extract and peptone) if the medium was supplemented with high concentrations of selenium. Concentrations of Na(2)SeO(3) that supported growth ranged from 3 to 100 mM. After 24 h of growth, the culture developed a deep red color characteristic of elemental selenium. When selenium was provided in the form of selenate, the pattern of growth showed a prolonged lag period, from 24 to 48 h. Final growth remained below that of cells cultured in the presence of selenite, and only a light red color developed. Concentrations of selenate below 40 mM failed to support growth. Tellurate, though not tellurite, could replace selenite, but only over a narrow concentration range, 5 to 10 mM. By 24 h, the typical black color of elemental tellurium developed. Bacillus sp. strain SS grew also in brain heart infusion broth and Trypticase soy broth (BBL Microbiology Systems, Cockeysville, Md.) without the addition of selenium or tellurium compounds. When added to these media, 50 mM selenite was tolerated and metabolized by the organism. The crucial distinction between this bacillus and other selenium-tolerant organisms (e.g., Salmonella) remains: under certain conditions, growth requirements of Bacillus sp. strain SS are fulfilled by selenium (and tellurium) compounds. PMID- 16346101 TI - Selective Isolation of Fusarium graminearum Mutants Derepressed for Production of beta-Glucosidase. AB - A strain of Fusarium graminearum produced extracellular beta-glucosidase (beta-d glucoside glucohydrolase [EC 3.2.1.21]) subject to carbon catabolite repression. Derepressed mutants were selectively isolated by the use of 2-deoxyglucose, a nonmetabolizable catabolic repressor. On each plate of cellobiose and 2-deoxy glucose inoculated with 10 spores and irradiated with UV light, a few colonies emerged. Comparative growth experiments with one of the derepressed mutants and the parent strain showed that the mutant produced beta-glucosidase in the presence of glucose. Furthermore, the same mutant produced more beta-glucosidase than did the wild-type strain on cellobiose alone. PMID- 16346102 TI - Improved method for detection of starch hydrolysis. AB - A new starch hydrolysis detection method which does not rely on iodine staining or the use of color-complexed starch is described. A linear relationship was obtained with agar-starch plates when net clearing zones around colonies of yeasts were plotted against enzyme levels (semilogarithm scale) produced by the same yeast strains in liquid medium. A similar relationship between starch clearing zones and alpha-amylase levels from three different sources was observed. These observations suggest that the method is useful in mutant isolations, strain improvement programs, and the prediction of alpha-amylase activities in culture filtrates or column effluents. PMID- 16346103 TI - Kaolin powder as a fungal carrier. AB - Kaolin powder may be used to carry and preserve an Aspergillus sp. for at least 90 days without contamination and with retention of its activity in softening and upgrading barky jute. PMID- 16346104 TI - Isolation of marine bacteria capable of producing specific lyases for alginate degradation. AB - Alginate lyases (EC 4.2.2.3) were isolated from cultures of several marine bacterial isolates. The lyases were induced by native alginate and had activity toward both the mannuronic acid and the guluronic acid blocks of the alginate polymer. The guluronic acid-specific lyase was recovered from the medium, whereas the mannuronic acid-specific lyase was retained with the bacteria. PMID- 16346106 TI - Sulfate reduction and methanogenesis in the sediment of a saltmarsh on the East coast of the United kingdom. AB - [This corrects the article on p. 989 in vol. 43.]. PMID- 16346105 TI - Inactivation of Glutamine Synthetase by Ammonia Shock in the Gram-Positive Bacterium Streptomyces cattleya. AB - In cultures of the gram-positive bacterium Streptomyces cattleya, a rapid inactivation of glutamine synthetase was seen after ammonia shock. pH activity curves for ammonia-shocked and control cultures are shown. A peak of glutamine synthetase activity was seen during fermentation for production of the antibiotic thienamycin. PMID- 16346107 TI - Enhanced Production of 2,3-Butanediol by Klebsiella pneumoniae Grown on High Sugar Concentrations in the Presence of Acetic Acid. AB - The bioconversion of sugars present in wood hemicellulose to 2,3-butanediol (hereafter referred to as butanediol) by Klebsiella pneumoniae grown on high initial concentrations (up to 10%) of sugars was investigated. Initial fermentation studies with a chemically defined medium suggested that sugar levels in excess of 2% could not be utlized even when a higher inoculum size (5 to 10%) was used. The addition of nutrient supplements, viz., yeast extract, urea, ammonium sulfate, and trace elements resulted in a 10 to 50% increase in butanediol yields, although sugar utilization remained incomplete. The concentration of end products normally found at the termination of fermentation was shown to be noninhibitory to growth and substrate utilization. Acetic acid was inhibitory at concentrations above 1%, although growth and butanediol yield were stimulated in cultures supplemented with lower levels of acetic acid. The efficient utilization of 4% substrate concentrations of d-glucose and d-xylose was achieved, resulting in butanediol yields of 19.6 and 22.0 g/liter, respectively. PMID- 16346108 TI - Initial Stages in the Biodegradation of the Surfactant Sodium Dodecyltriethoxy Sulfate by Pseudomonas sp. Strain DES1. AB - The biodegradation of the surfactant sodium dodecyltriethoxy sulfate by Pseudomonas sp., strain DES1 (isolated from activated sludge plant effluent) has been studied. Growth of the organism when the S-labeled surfactant was present as the sole source of carbon and energy led to the appearance in the culture fluid of five S-labeled organic metabolites. These have been identified as mono-, di-, and triethylene glycol monosulfates (major metabolites) and acetic acid 2-(ethoxy sulfate) and acetic acid 2-(diethoxy sulfate), authentic samples of which have been prepared and characterized. Evidence is presented that the major metabolites were produced by rupture of one or another of the three ether linkages present in the surfactant molecule, probably via the agency of a single etherase enzyme. Acetic acid 2-(ethoxy sulfate) and acetic acid 2-(diethoxy sulfate) were formed by the oxidation of the free alcohol groups of di- and triethylene glycol monosulfates, respectively, and increased in amount during the stationary phase of growth. Inorganic S-sulfate also appeared in significant quantities in culture fluids and arose from the parent surfactant (presumably via the action of an alkylsulfatase) and not from any of the five metabolites. The appearance of sulfated organic metabolites during the exponential phase of growth and their quantitative relationship remained remarkably constant, even when additional carbon and energy sources (succinate or yeast extract) were also present in the growth media. PMID- 16346109 TI - Temperature adaptations in the terminal processes of anaerobic decomposition of yellowstone national park and icelandic hot spring microbial mats. AB - The optimum temperatures for methanogenesis in microbial mats of four neutral to alkaline, low-sulfate hot springs in Yellowstone National Park were between 50 and 60 degrees C, which was 13 to 23 degrees C lower than the upper temperature for mat development. Significant methanogenesis at 65 degrees C was only observed in one of the springs. Methane production in samples collected at a 51 or 62 degrees C site in Octopus Spring was increased by incubation at higher temperatures and was maximal at 70 degrees C. Strains of Methanobacterium thermoautotrophicum were isolated from 50, 55, 60, and 65 degrees C sites in Octopus Spring at the temperatures of the collection sites. The optimum temperature for growth and methanogenesis of each isolate was 65 degrees C. Similar results were found for the potential rate of sulfate reduction in an Icelandic hot spring microbial mat in which sulfate reduction dominated methane production as a terminal process in anaerobic decomposition. The potential rate of sulfate reduction along the thermal gradient of the mat was greatest at 50 degrees C, but incubation at 60 degrees C of the samples obtained at 50 degrees C increased the rate. Adaptation to different mat temperatures, common among various microorganisms and processes in the mats, did not appear to occur in the processes and microorganisms which terminate the anaerobic food chain. Other factors must explain why the maximal rates of these processes are restricted to moderate temperatures of the mat ecosystem. PMID- 16346110 TI - Chemical and Physical Characterization of Interfacial-Active Lipids from Rhodococcus erythropolis Grown on n-Alkanes. AB - Lipophilic compounds of the culture suspension containing Rhodococcus erythropolis DSM43215 had surfactant properties when the bacteria were cultivated with n-alkanes as the sole carbon source. Thirteen main components from a dichloromethane-methanol extract of the R. erythropolis cultures were isolated and characterized to specify quantitatively their surfactant properties, e.g., minimum surface and interfacial tensions and critical micelle concentrations. The interfacial activity of the organic extract was dominated by alpha,alpha trehalose-6,6'-dicorynomycolates which reduced interfacial tension from 44 to 18 mN/m. Phosphatidylethanolamines which were also present in the organic extract reduced the interfacial tension below 1 mN/m. The trehalose corynomycolates had extremely low critical micelle concentrations in high-salinity solutions, and the interfacial properties were stabile in solutions with a wide range of pH and ionic strength. PMID- 16346111 TI - Bacterial nitration of 4-chlorobiphenyl. AB - In the course of a study dealing with the biodegradation of 4-chlorobiphenyl by strain B-206, we noticed that the gram-negative bacterium accumulated different metabolic intermediates depending on the nitrogen source of the medium. Hence, in the presence of nitrate, strain B-206 produced four compounds which were identified as 2- and 4-hydroxy-4'-chlorobiphenyl and 2- and 4-hydroxy-mononitro 4'-chlorobiphenyl. The accumulation of these compounds in the culture medium indicated the presence of a monooxygenase in strain B-206 leading to the production of arene oxide intermediates. The possible transformation of 4 chlorobiphenyl to an arene oxide by this bacterial strain is a matter of concern because of the high reactivity of these arene oxides with biological material. PMID- 16346112 TI - Removal of Sulfur Compounds from Coal by the Thermophilic Organism Sulfolobus acidocaldarius. AB - The thermophilic, reduced-sulfur, iron-oxidizing bacterium Sulfolobus acidocaldarius was used for the removal of sulfur compounds from coal. The inclusion of complex nutrients such as yeast extract and peptone, and chemical oxidizing agents, 0.01 M FeCl(3) into leaching medium, reduced the rate and the extent of sulfur removal from coal. The rate of sulfur removal by S. acidocaldarius was strongly dependent on the sulfur content of the coal and on the total external surface area of coal particles. Approximately 96% of inorganic sulfur was removed from a 5% slurry of coal which had an initial total sulfur content of 4% and an inorganic (pyritic S and sulfate) sulfur content of 2.1%. This resulted in removal of 50% of initial total sulfur present in coal. PMID- 16346113 TI - Effect of salinity on Rhizobium growth and survival. AB - This study examines the effect of salinity on the growth and survival of Rhizobium spp. in culture media and soil. Eleven isolates from saline and nonsaline environments were compared. The growth (mean doubling time) of all strains and species tested decreased when the electrical conductivity of the culture medium (yeast extract-mannitol) was raised from 1.2 mS cm to 6.7 mS cm (15% seawater equivalent) or to 13.1 mS cm (28% seawater equivalent). Three of eleven strains failed to grow at 13.1 mS cm. Although growth was affected by salinity, four strains selected from the growth rate study could survive in extremely high concentrations of salt. Two strains with growth rates sensitive to salt and two strains with growth rates relatively unaffected by salt were inoculated into solutions with electrical conductivities of up to 43.0 mS cm (92% seawater equivalent). Not only did all four strains survive the initial osmotic shock (at 5 h after inoculation), but it was not until 27 days after inoculation that the sensitive strains exhibited a significant reduction in viable numbers. The salt-tolerant strains survived for more than 65 days with no reduction in viable counts. The interaction between soil moisture tension and soil salinity in relation to Rhizobium survival in gamma-irradiated soil was also examined. Six treatment combinations were used, ranging from -0.1 bars and 0.2 mS cm to -15 bars and 12 mS cm. Sensitive strains declined from 10 to 10 organisms per g of soil after 84 days of incubation at -15 bars and 12 mS cm. Tolerant strains survived for the same period with no loss in viable numbers. The results of these experiments indicate that many strains of Rhizobium can grow and survive at salt concentrations which are inhibitory to most agricultural legumes. The emphasis of research concerning the effects of salinity on symbiotic nitrogen fixation should, therefore, be directed to aspects of the symbiosis other than the survival of the Rhizobium spp. PMID- 16346114 TI - Selected nucleic Acid precursors in studies of aquatic microbial ecology. AB - The use of radiolabeled nucleosides and nucleic acid bases to estimate the rates of RNA and DNA synthesis in naturally occurring microbial assemblages requires numerous assumptions, several of which are evaluated herein. Comparative time series analyses of the uptake and incorporation, labeling specificity, and extent of catabolism of [2-H]adenine, [methyl-H]thymidine, and [5-H]uridine were performed with pure bacterial and algal cultures, as well as with environmental samples. [H]thymidine yielded the most variable results, especially with regard to the extent of nonspecific macromolecular labeling. The pathways of [H]thymidine and [H]adenine metabolism were further evaluated by isotope dilution methods and by comparing incorporation patterns of thymidine labeled at different sites of the molecule. The advantages, uncertainties, and limitations of the use of radiolabeled nucleic acid precursors in studies of aquatic microbial ecology are discussed and a prospectus for future studies presented. PMID- 16346115 TI - Concurrent Production and Consumption of Ethanol by Cultures of Pachysolen tannophilus Growing on d-Xylose. AB - Growing cultures of Pachysolen tannophilus concurrently consumed and produced ethanol in the presence of substantial concentrations of d-xylose. Ethanol was also assimilated in the presence of other sugars, the amount depending on the sugar. Less ethanol assimilation occurred with d-glucose than with d-xylose. The rate of ethanol consumption decreased as the concentration of glucose was increased, but some consumption still occurred when 2% glucose was present. The rate increased with the amount of oxygen available to the culture when d-xylose or ethanol was the carbon source. In most instances, estimates of consumption were based on the extent of incorporation of C from [1-C]ethanol into trichloroacetic acid-insoluble material. The results are pertinent to the use of P. tannophilus for the production of ethanol from d-xylose. PMID- 16346116 TI - Changes in Viability, Cell Composition, and Enzyme Levels During Starvation of Continuously Cultured (Ammonia-Limited) Selenomonas ruminantium. AB - Under nitrogen (ammonia)-limited continuous culture conditions, the ruminal anaerobe Selenomonas ruminantium was grown at various dilution rates (D). The proportion of the population that was viable increased with D, being 91% at D = 0.5 h. Washed cell suspensions were subjected to long-term nutrient starvation at 39 degrees C. All populations exhibited logarithmic linear declines in viability that were related to the growth rate. Cells grown at D = 0.05, 0.20, and 0.50 lost about 50% viability after 8.1, 4.6, and 3.6 h, respectively. The linear rates of decline in total cell numbers were dramatically less and constant regardless of dilution rate. All major cell constituents declined during starvation, with the rates of decline being greatest with RNA, followed by DNA, carbohydrate, cell dry weight, and protein. The rates of RNA loss increased with cells grown at higher D values, whereas the opposite was observed for rates of carbohydrate losses. The majority of the degraded RNA was not catabolized but was excreted into the suspending buffer. At all D values, S. ruminantium produced mainly lactate and lesser amounts of acetate, propionate, and succinate during growth. With starvation, only small amounts of acetate were produced. Addition of glucose, vitamins, or both to the suspending buffer or starvation in the spent culture medium resulted in greater losses of viability than in buffer alone. Examination of extracts made from starving cells indicated that fructose diphosphate aldolase and lactate dehydrogenase activities remained relatively constant. Both urease and glutamate dehydrogenase activities declined gradually during starvation, whereas glutamine synthetase activity increased slightly. The data indicate that nitrogen (ammonia)-limited S. ruminantium cells have limited survival capacity, but this capacity is greater than that found previously with energy (glucose)-limited cells. Apparently no one cellular constituent serves as a catabolic substrate for endogenous metabolism. Relative to losses in viability, cellular enzymes are stable, indicating that nonviable cells maintain potential metabolic activity and that generalized, nonspecific enzyme degradation is not a major factor contributing to viability loss. PMID- 16346117 TI - Nitrogen Redox Metabolism of a Heterotrophic, Nitrifying-Denitrifying Alcaligenes sp. from Soil. AB - Metabolic characteristics of a heterotrophic, nitrifier-denitrifier Alcaligenes sp. isolated from soil were further characterized. Pyruvic oxime and hydroxylamine were oxidized to nitrite aerobically by nitrification-adapted cells with specific activities (V(max)) of 0.066 and 0.003 mumol of N x min x mg of protein, respectively, at 22 degrees C. K(m) values were 15 and 42 muM for pyruvic oxime and hydroxylamine, respectively. The greater pyruvic oxime oxidation activity relative to hydroxylamine oxidation activity indicates that pyruvic oxime was a specific substrate and was not oxidized appreciably via its hydrolysis product, hydroxylamine. When grown as a denitrifier on nitrate, the bacterium could not aerobically oxidize pyruvic oxime or hydroxylamine to nitrite. However, hydroxylamine was converted to nearly equimolar amounts of ammonium ion and nitrous oxide, and the nature of this reaction is discussed. Cells grown as heterotrophic nitrifiers on pyruvic oxime contained two enzymes of denitrification, nitrate reductase and nitric oxide reductase. The nitrate reductase was the dissimilatory type, as evidenced by its extreme sensitivity to inhibition by azide and by its ability to be reversibly inhibited by oxygen. Cells grown aerobically on organic carbon sources other than pyruvic oxime contained none of the denitrifying enzymes surveyed but were able to oxidize pyruvic oxime to nitrite and reduce hydroxylamine to ammonium ion. PMID- 16346118 TI - Role of Thin Fimbriae in Adherence and Growth of Acinetobacter calcoaceticus RAG 1 on Hexadecane. AB - Acinetobacter calcoaceticus RAG-1, a hydrocarbon-degrading bacterium which adheres avidly to hydrocarbons and other hydrophobic surfaces, possesses numerous thin fimbriae (ca. 3.5-nm diameter) on the cell surface. MR-481, a nonadherent mutant of RAG-1 which is unable to grow on hexadecane under conditions of limited emulsification and low initial cell density, lacks these fimbriae. Prolonged incubation of MR-481 in hexadecane medium enriched for partial adherence revertants. The reappearance of thin fimbriae was observed in all such revertant strains. RAG-1 cells and partial revertant strains were agglutinated in the presence of antibody, whereas MR-481 cells were not. Another mutant, AB15, which was previously isolated on the basis of its nonagglutinability in the presence of antibody, also lacked thin fimbriae and was conditionally nonadherent. Furthermore, strain AB15 was unable to grow on hexadecane medium. Adherence of RAG-1 cells to hexadecane was considerably reduced after shearing treatment. The material removed from the cell surface by shearing of RAG-1 and the partial revertant strains yielded a single antigenic band in RAG-1 and partial revertant strains, as observed by crossed immunoelectrophoresis. This band was absent in both fimbriae-less mutants, MR-481 and AB15. The data demonstrate that the thin fimbriae of RAG-1 (i) are a major factor in adherence to polystyrene and hydrocarbon, (ii) may be crucial in enabling growth of cells on hexadecane, and (iii) constitute the major cell surface agglutinogen. PMID- 16346119 TI - Adaptation to Cadmium by Klebsiella aerogenes Growing in Continuous Culture Proceeds Mainly via Formation of Cadmium Sulfide. AB - The adaptation of Klebsiella aerogenes to high levels of cadmium was studied in continuous culture under conditions of glucose limitation. When up to 6 x 10 M cadmium was added to a culture in steady state, growth ceased instantaneously but resumed within 5 h (dilution rate, 0.1 h). When again in steady state, these adapted cells exhibited a far greater tolerance to cadmium than did unadapted cells (not previously exposed to cadmium) when tested on solid media containing different concentrations of cadmium. This relative insensitivity of adapted cells to cadmium was subsequently lost in continuous culture within 5 days after omitting cadmium from the influent medium. Thus, the phenomenon was an inducible physiological process. Adapted cells contained substantial amounts of cadmium (up to 2.4% of the bacterial dry weight). The cadmium content of the cells was dependent on growth conditions and was found to be proportional to the inorganic sulfide content of the cells in all cases. This suggested that formation of CdS is probably the most important mechanism of detoxification in this organism. The presence of large numbers of electron-dense granules on the cell surface (absent in cultures without added cadmium) provided additional support for this conclusion. PMID- 16346120 TI - Improved microautoradiographic method to determine individual microorganisms active in substrate uptake in natural waters. AB - We report a method which combines epifluorescence microscopy and microautoradiography to determine both the total number of microorganisms in natural water populations and those individual organisms active in the uptake of specific substrates. After incubation with H-labeled substrate, the sample is filtered and, while still on the filter, mounted directly in a film of autoradiographic emulsion on a microscope slide. The microautoradiogram is processed and stained with acridine orange, and, subsequently, the filter is removed before microscopic observation. This novel preparation resulted in increased accuracy in direct counts made from the autoradiogram, improved sensitivity in the recognition of uptake-active (H-labeled) organisms, and enumeration of a significantly greater number of labeled organisms compared with corresponding samples prepared by a previously reported method. PMID- 16346121 TI - Nodulation of Pole Bean (Phaseolus vulgaris L.) by Rhizobium Species of Two Cross Inoculation Groups. AB - Physiology and morphology of pole bean (Phaseolus vulgaris L. cv Kentucky Wonder) root nodules induced by two Rhizobium species of different cross-inoculation groups have been compared. Root nodules induced by Rhizobium sp. 127E15, which is a strain of the cowpea group Rhizobium, were pinkish, had irregular shapes, and were only partially effective. Their peak acetylene reduction activity was 4.36 mumol of C(2)H(4) formed per g of fresh nodules per h at 30 days after inoculation. The effective nodules induced by Rhizobium phaseoli 127K14, which is a strain of the bean group Rhizobium, were dark red, spherical, and showed peak acetylene reduction activity of 15.95 mumol of C(2)H(4) formed per g of fresh nodules per h at 15 days after inoculation. The partial effectiveness of 127E15 induced nodules was associated with fewer infected cells, a delay in the increase of bacteroid population within the host cells, abundance of cytoplasmic vesicles in the host cells, more bacteroids within a membrane envelope (peribacteroid membrane), and the inability of bacteroids to completely fill up the host cytoplasmic space. The 127K14-induced nodules were fully mature, with host cells filled with bacteroids by 12 days after inoculation. In contrast, the 127E15 induced nodules did not reach a similar developmental stage even 30 days after inoculation. PMID- 16346122 TI - Fungal invertase as an aid for fermentation of cane molasses into ethanol. AB - Comparative studies of the fermentation of cane molasses into ethanol by Saccharomyces cerevisiae in the presence or absence of fungal invertase were performed. When cane molasses was fermented by the yeast at 30 degrees C and pH 5.0, the presence of the enzyme had no effect on ethanol production. At pH 3.5, ethanol production was increased by the addition of invertase. At 40 degrees C, the addition of invertase increased ethanol production by 5.5% at pH 5.0 and by 20.9% at pH 3.5. PMID- 16346123 TI - Improved Medium for Isolation of Azospirillum spp. AB - Colonies of Azospirillum spp. could be readily distinguished from colonies of other diazotrophs by scarlet coloration in culture media in which Congo red was included. PMID- 16346124 TI - Isolation and Complementation Studies of Auxotrophic Mutants of the Lignin Degrading Basidiomycete Phanerochaete chrysosporium. AB - A variety of auxotrophic strains of Phanerochaete chrysosporium were isolated after treatment of conidia with UV and X rays. Complementation studies with these strains demonstrated heterokaryotic mycelia and conidia in this organism. Nuclear staining also showed that conidia can be mono-, di-, or multinucleate. Complementation tests allowed the separation of each auxotrophic class with the same phenotype into complementation groups. PMID- 16346125 TI - Optimization of Conditions for Photoproduction of Ammonia from Nitrate by Anacystis nidulans. AB - The effect of several relevant environmental factors influencing the photoproduction of ammonia from nitrate by Anacystis nidulans cells treated with the glutamine synthetase inhibitor l-methionine-dl-sulfoximine has been investigated. The optimal ratio between l-methionine-dl-sulfoximine concentration (micro-molar) and cell density (micrograms of chlorophyll per milliliter) was around 1, the process taking place at maximal rate at a temperature of about 40 degrees C, within the pH range of 7 to 10. Ammonia production was stimulated by CO(2) or bicarbonate and was not affected by the accumulation of ammonia in the medium up to concentrations of 30 mM. The rate of ammonia production was found to be determined by the interaction of at least four factors, namely, irradiance and the density, depth, and turbulence of the cell suspension. Ammonia photoproduction from nitrate and water represents an interesting process for the conversion of light energy into chemical energy, which can operate at high efficiency, around 30% of its theoretical maximum. PMID- 16346126 TI - Sustained Photoproduction of Ammonia from Nitrate by Anacystis nidulans. AB - Conditions that lengthen the time during which l-methionine-dl-sulfoximine (MSX) promotes excretion of ammonia produced by photosynthetic nitrate reduction in Anacystis nidulans have been sought. If MSX was added every 24 h, maximal rates of ammonia production were maintained for 3 days. After this time, ammonia production ceased due to a specific deficiency of glutamine in the cells, which finally led to cell lysis. The effective ammonia production period could be further extended either by adding a low amount of glutamine at the end of the 3 day period or by allowing the cells to recover for 8 h in the absence of MSX after every 48-h period in the presence of inhibitor. In this way, a steady production of ammonia lasting for at least 10 days was achieved. The MSX-treated cyanobacterial cells thus represent a system relatively stable with time for the conversion of light energy into chemical energy through the photoreduction of nitrate to ammonia. PMID- 16346127 TI - Production of a Thermostable beta-d-Galactosidase by Alternaria alternata Grown in Whey. AB - In the course of exploring new microbial sources of extracellular beta-d galactosidase (EC. 3.2.1.23), Alternaria alternata was found to excrete elevated quantities of a thermostable form of the enzyme when cultivated in whey growth medium. Optimum cultural conditions for maximum enzyme production were a whey lactose concentration of 6%, supplementation of the medium with 0.050 M (NH(4))(2)SO(4), an inoculum size of 10 conidia per ml, and a cultivation time at 28 to 30 degrees C of 5 days. The fungus utilized whey lactose for the production of the enzyme most efficiently, and the observed maximum yield, 280 nanokatals of hydrolyzed o-nitrophenyl-beta-d-galactopyranoside per g of whey lactose, was comparable to maximum yields reported for certain commercial fungi. The optimum pH and temperature of the enzymatic reaction were 4.5 to 5.5 and 60 to 70 degrees C, respectively, and the enzyme lost half of its activity when heated at 65 degrees C for 84 min. These properties make the enzyme particularly suitable for processing acid and less-acid (pH 5 to 6) dairy products and by-products. PMID- 16346128 TI - Direct Fermentation of d-Xylose to Ethanol by Kluyveromyces marxianus Strains. AB - Eight strains of Kluyveromyces marxianus were screened, and all of them were found to ferment the aldopentose d-xylose directly to ethanol under aerobic conditions. One of these strains, K. marxianus SUB-80-S, was grown in a medium containing 20 g of d-xylose per liter, and the following results were obtained: maximum ethanol concentration, 5.6 g/liter; ethanol yield, 0.28 g of ethanol per g of d-xylose (55% of theoretical); maximum specific growth rate, 0.12 h; 100% d xylose utilization was completed in 48 h. PMID- 16346129 TI - Plants as sources of airborne bacteria, including ice nucleation-active bacteria. AB - Vertical wind shear and concentration gradients of viable, airborne bacteria were used to calculate the upward flux of viable cells above bare soil and canopies of several crops. Concentrations at soil or canopy height varied from 46 colony forming units per m over young corn and wet soil to 663 colony-forming units per m over dry soil and 6,500 colony-forming units per m over a closed wheat canopy. In simultaneous samples, concentrations of viable bacteria in the air 10 m inside an alfalfa field were fourfold higher than those over a field with dry, bare soil immediately upwind. The upward flux of viable bacteria over alfalfa was three- to fourfold greater than over dry soil. Concentrations of ice nucleation-active bacteria were higher over plants than over soil. Thus, plant canopies may constitute a major source of bacteria, including ice nucleation-active bacteria, in the air. PMID- 16346130 TI - Competitiveness of Rhizobium trifolii Strains Associated with Red Clover (Trifolium pratense L.) in Mississippi Soils. AB - Five strains of Rhizobium trifolii were evaluated in competition with indigenous populations in nodulating red clover (Trifolium pratense L.) cv. Kenland in two different soils in Mississippi. Double antibiotic resistance acquisition was used to measure the proportion of nodules occupied by the introduced mutant strains. In vertisol soil, strains RP113-7, 162BB1, LM1, and 162P17 were recovered in at least 94% of the assayed nodules, whereas TA1 was found in 83.8% of the nodules. At an ultisol location, significant differences were detected within the introduced rhizobia. Strain RP113-7 was recovered at very high rates (99.2% of the assayed nodules), whereas strains 162BB1, LM1, 162P17, and TA1 were all found in 84.9 to 96.0% of the nodules sampled. Forage yield and percent crude protein levels were lower with the less effective but competitive strain (TA1) at both locations. Results indicated that more effective strains of R. trifolii can increase red clover production and symbiotic nitrogen fixation under different environmental conditions in Mississippi. PMID- 16346131 TI - Microcalorimetric Measurements of Glucose Metabolism by Marine Bacterium Vibrio alginolyticus. AB - Microcalorimetric measurements of heat production from glucose by Vibrio alginolyticus were made to assess the viability of calorimetry as a technique for studying the metabolism of marine bacteria at organic nutrient concentrations found in marine waters. The results show that the metabolism of glucose by this bacterium can be measured by calorimetry at submicromolar concentrations. A linear correlation between glucose concentration and total heat production was observed over a concentration range of 8 mM to 0.35 muM. It is suggested that these data indicate a constant efficiency of metabolism for this bacterium over the wide range of glucose concentrations studied. PMID- 16346132 TI - Impact of coal-coking effluent on sediment microbial communities: a multivariate approach. AB - The functional response to and recovery from coal-coking waste effluent was evaluated for sediment microbial communities. Twenty estimates of microbial population density, biomass, and activity were measured five times during a 15 month period. Significant effects on microbial communities were observed in response to both wastewater contamination and diversion of the wastewater. Multivariate analysis of variance and discriminant analysis indicated that accurate differentiation between uncontaminated and contaminated sediments required a minimum of nine estimates of community response. Total viable population density, ATP, alkaline phosphatase, naphthalene, and phenanthrene mineralization rates were found to be highly weighted variables in site discrimination. Lipid and glucose mineralization, nitrogen fixation, and sediment protein also contributed significantly to explaining variation among sites. Estimates of anaerobic population densities and rates of methane production contributed little to discrimination among sites in the environment examined. In general, total viable population density, ATP, and alkaline phosphatase activity were significantly depressed in contaminated sediments. However, after removal of this contamination, the previously affected sites demonstrated greater temporal variability but a closer approximation of the mean response at the control site. Naphthalene and phenanthrene mineralization did not follow the general trend and were elevated at the contaminated sites throughout the investigation. Results of the investigation supported the hypothesis that multiple functional measures of microbial community response are required to evaluate the effect of and recovery from environmental contamination. In addition, when long-term effects are evaluated, select physiological traits, i.e., polyaromatic hydrocarbon mineralization, may not reflect population and biomass estimates of community response. PMID- 16346133 TI - Microbial Oxidation of Hydrocarbons: Properties of a Soluble Methane Monooxygenase from a Facultative Methane-Utilizing Organism, Methylobacterium sp. Strain CRL-26. AB - Methylobacterium sp. strain CRL-26 grown in a fermentor contained methane monooxygenase activity in soluble fractions. Soluble methane monooxygenase catalyzed the epoxidation/hydroxylation of a variety of hydrocarbons, including terminal alkenes, internal alkenes, substituted alkenes, branched-chain alkenes, alkanes (C(1) to C(8)), substituted alkanes, branched-chain alkanes, carbon monoxide, ethers, and cyclic and aromatic compounds. The optimum pH and temperature for the epoxidation of propylene by soluble methane monooxygenase were found to be 7.0 and 40 degrees C, respectively. Among various compounds tested, only NADH(2) or NADPH(2) could act as an electron donor. Formate and NAD (in the presence of formate dehydrogenase contained in the soluble fraction) or 2 butanol in the presence of NAD and secondary alcohol dehydrogenase generated the NADH(2) required for the methane monooxygenase. Epoxidation of propylene catalyzed by methane monooxygenase was not inhibited by a range of potential inhibitors, including metal-chelating compounds and potassium cyanide. Sulfhydryl agents and acriflavin inhibited monooxygenase activity. Soluble methane monooxygenase was resolved into three components by ion-exchange chromatography. All three compounds are required for the epoxidation and hydroxylation reactions. PMID- 16346134 TI - Preservation of estuarine sediments for lipid analysis of biomass and community structure of microbiota. AB - Various methods were tested for preserving estuarine sediments in the field before biochemical analysis of the microbiota. Total microbial biomass was determined as lipid phosphate (LP), and the fatty acids of the microbial lipids were used as indicators of community structure. Control samples were sieved to remove macroinvertebrates and plant materials and were extracted immediately in the field. Other samples were preserved both before and after sieving and stored for 5 days before analysis. Freezing resulted in a 50% decline in LP and significant decreases in many fatty acids. Refrigeration resulted in a 19% decrease in LP but no change in the fatty acids. Samples preserved with Formalin before sieving exhibited no significant change in LP but substantial increases in many fatty acids, which were probably derived from the macroinvertebrates. Sieved samples preserved with Formalin showed a 17 to 18% decline in LP but no change in the fatty acids. Ideally, samples should be sieved and extracted immediately in the field. However, short-term refrigeration and longer-term preservation of sieved samples with Formalin may be acceptable compromises. PMID- 16346135 TI - Optimal Cultural and Physiological Conditions for Handling Streptomyces rimosus Protoplasts. AB - A general procedure for manipulating protoplasts of three Streptomyces rimosus strains was developed. More than 50% regeneration efficiency was obtained by optimizing the osmotic stabilizer concentrations and modifying the plating procedure. Preparation and regeneration of protoplasts were studied by both phase contrast and electron microscopy. After cell wall degradation with lysozyme, protoplasts about 1,000 to 1,500 nm in diameter appeared. The reversion process exhibited normal and aberrant regeneration of protoplasts to hyphae and to spherical cells, respectively. Spherical cells contained no alpha, epsilon-ll diaminopimelic acid and were colorless or red after Gram staining. They showed consistent stability during at least five subsequent subcultivations. However, the omission of glycine from the precultivation medium reduced the unusual process of regeneration almost completely. After normal protoplast regeneration, the production of oxytetracycline by single isolates was not affected. PMID- 16346136 TI - Factors regulating microbial biofilm development in a system with slowly flowing seawater. AB - Microbial biofilm development was followed under growth conditions similar to those of a projected salinity power plant. Microscope glass cover slips were piled in biofilm reactors to imitate the membrane stacks in such a plant. A staining technique closely correlating absorbance values with biofilm dry weight was used for the study. Generally, the biofilms consisted of solitary and filamentous bacteria which were evenly distributed with considerable amounts of various protozoa and entrapped debris of organic origin. Protozoa predation was shown to decrease the amount of biofilm produced. The biofilm development lag phase was longer at lower temperatures. The subsequent growth phase was approximately arithmetic until stationary phase appeared. Adaptation of a hyperbolic saturation function gave curves that agreed well with the logarithm of the amount of biofilm as a function of time. Increased flow velocity, temperature, and nutrient concentration increased the biofilm production rate. An exponential relationship was shown between biofilm production rate and flow velocity within the range of 0 to 15 cm s. Intervals in which the biofilms were exposed to fresh water decreased the biofilm production rate more than four times. If the cover slips were inoculated with untreated seawater for 24 h, subsequent UV treatment had an insignificant effect on the biofilm formation. PMID- 16346137 TI - Barophilic bacteria associated with digestive tracts of abyssal holothurians. AB - Abyssal holothurians and sediment samples were collected at depths of 4,430 to 4,850 m in the Demerara abyssal plain. Bacterial concentrations in progressive sections of the holothurian digestive tract, as well as in surrounding surface sediments, were determined by epifluorescence microscopy. Total bacterial counts in sediments recently ingested by the animals were 1.5- to 3-fold higher than in surrounding sediments at the deepest station. Lowest counts were observed consistently in the foregut, where the digestive processes of the holothurian are believed to occur. In most animals, counts increased 3- to 10-fold in the hindgut. Microbial activity at 3 degrees C and in situ and atmospheric pressure were determined for gut and sediment samples by measuring the utilization of [C]glutamic acid, the doubling time of the mixed-population of culturable bacteria, and the percentage of the total bacterial count responsive to yeast extract in the presence of nalidixic acid, using epifluorescence microscopy. A barophilic microbial population, showing elevated activity under deep-sea pressure, was detected by all three methods in sediments removed from the hindgut. Transmission electron micrographs revealed intact bacteria directly associated with the intestinal lining only in the hindgut. The bacteria are believed to be carried as an actively metabolizing, commensal gut flora that transforms organic matter present in abyssal sediments ingested by the holothurian. Using data obtained in this study, it was calculated that sediment containing organic matter altered by microbial activity cleared the holothurian gut every 16 h, suggesting that abyssal holothurians and their associated gut flora are important participants in nutrient cycles of the abyssal benthic ocean. PMID- 16346138 TI - Production of Extracellular Polysaccharide by Zoogloea ramigera. AB - In batch cultures of Zoogloea ramigera the maximum rate of exopolysaccharide synthesis occurred in a partly growth-linked process. The exopolysaccharide was attached to the cells as a capsule. The capsules were released from the cell walls after 150 h of cultivation, which caused the fermentation broth to be highly viscous. Ultrasonication could be used to release capsular polysaccharide from the microbial cell walls. Treatment performed after 48 to 66 h of cultivation revealed exopolysaccharide concentration and apparent viscosity values in accordance with values of untreated samples withdrawn after 161 h of cultivation. The yield coefficient of exopolysaccharide on the basis of consumed glucose was in the range of 55 to 60% for batch cultivations with an initial glucose concentration of 25 g liter. An exopolysaccharide concentration of up to 38 g liter could be attained if glucose, nitrogen, and growth factors were fed into the batch culture. The oxygen consumption rate in batch fermentations reached 25 mmol of O(2) liter h during the exopolysaccharide synthesis phase and then decreased to values below 5 mmol of O(2) liter h during the release phase. The fermentation broth showed pseudoplastic flow behavior, and the polysaccharide was not degraded when growth had ceased. PMID- 16346139 TI - Strain-specific differences in surface antigens of symbiotic algae. AB - Intracellular symbiotic dinoflagellates (zooxanthellae) isolated from different invertebrate hosts are morphologically similar but could be differentiated by the fluorescent-antibody technique. PMID- 16346140 TI - Ammonium production by dissimilatory nitrate reducers isolated from baltic sea water, as indicated by N study. AB - Bacteria able to perform dissimilatory nitrate reduction to ammonium were isolated from low-oxygen masses in the Baltic Sea. In liquid media enriched with NO(3) and incubated anaerobically, the NH(4)-producing isolates transformed 25 to 72% of the NO(3) to NH(4). PMID- 16346141 TI - Comparison of the Pour, Spread, and Drop Plate Methods for Enumeration of Rhizobium spp. in Inoculants Made from Presterilized Peat. AB - Inoculants prepared with presterilized peat were enumerated by the pour, spread, and drop plate techniques. Results indicated that the three plating methods were interchangeable. The drop plate method was preferred because of its economy in materials and labor. PMID- 16346142 TI - Microbial Production of Ursodeoxycholic Acid from Lithocholic Acid by Fusarium equiseti M41. AB - A fungus identified as Fusarium equiseti was isolated from soil and found to carry out 7beta-hydroxylation of lithocholic acid to ursodeoxycholic acid (35% yield; 350 mg/liter) in 112 h. PMID- 16346143 TI - Extracellular Isoamylase Produced by the Yeast Lipomyces kononenkoae. AB - A strain of the starch-converting yeast Lipomyces kononenkoae produced, when grown on starch, a debranching enzyme that proved to be an isoamylase (glycogen 6 glucanohydrolase; E.C. 3.2.1.68). So far, only bacteria have been found to produce extracellular isoamylases. The yeast isoamylase enhanced beta-amylolysis of amylopectin and glycogen and completely hydrolyzed these substrates into maltose when combined with a beta-amylase but had no action on dextran or pullulan. By isopropanol precipitation and carboxymethyl cellulose chromatography, L. kononenkoae isoamylase was partially purified from the supernatant of cultures grown on a mineral medium with soluble starch. Optimum temperature and pH for activity of the isoamylase were 30 degrees C and 5.6. The molecular weight was around 65,000, and the pI was at pH 4.7 to 4.8. The K(m) (30 degrees C, pH 5.5) for soluble starch was 9 g liter. PMID- 16346144 TI - Methanogenesis and sulfate reduction: competitive and noncompetitive substrates in estuarine sediments. AB - Sulfate ions did not inhibit methanogenesis in estuarine sediments supplemented with methanol, trimethylamine, or methionine. However, sulfate greatly retarded methanogenesis when hydrogen or acetate was the substrate. Sulfate reduction was stimulated by acetate, hydrogen, and acetate plus hydrogen, but not by methanol or trimethylamine. These results indicate that sulfate-reducing bacteria will outcompete methanogens for hydrogen, acetate, or both, but will not compete with methanogens for compounds like methanol, trimethylamine, or methionine, thereby allowing methanogenesis and sulfate reduction to operate simultaneously within anoxic, sulfate-containing sediments. PMID- 16346145 TI - Autolytic Activity and Butanol Tolerance of Clostridium acetobutylicum. AB - The effects of acetone and butanol on the growth of vegetative cells and the stability of swollen-phase bright-stationary-phase cells (clostridial forms) of Clostridium acetobutylicum P262 and an autolytic deficient mutant (lyt-1) were investigated. There was little difference in the sensitivity of strain P262 and the lyt-1 mutant vegetative cells and clostridial forms to acetone. The stability of the different morphological stages was unaffected by acetone concentrations far in excess of those encountered in factory fermentations. Butanol concentrations between 7 and 16 g/liter, which are within the range obtained in industrial fermentations, increased the degeneration of strain P262 clostridial forms but had no effect on the stability of lyt-1 clostridial forms which never underwent autolysis. Vegetative cells of the lyt-1 mutant were able to grow in higher concentrations of butanol than strain P262 vegetative cells. It was concluded that there is a relationship between butanol tolerance and autolytic activity. PMID- 16346146 TI - Influence of Temperature Adaptation on Glucose Metabolism in a Psychrotrophic Strain of Cytophaga johnsonae. AB - Selective enrichment of yellow-orange-pigmented, gram-negative bacteria related to Cytophaga johnsonae from lake sediment was dependent on low temperatures (ca. 5 degrees C). However, this temperature effect was abolished when excessive amounts of dissolved organic carbon (10 mM N-acetylglucosamine) were added. A psychrotrophic freshwater isolate of C. johnsonae was used to study the physiological versatility of this group. Exponential growth rates were found to be dependent on the temperature to which the cells used as the inocula were acclimated. Glucose incorporation and respiration were also dependent upon the acclimation temperature of the inocula. Patterns of CO(2) evolution obtained from position-labeled [C]glucose indicated that glucose was predominantly metabolized via the Embden-Meyerhof-Parnas pathway, which, however, was greatly reduced at 25 degrees C when the concentration of glucose was as low as 5 muM/liter. Transport, respiration, and incorporation of glucose (0.2- to 20,000-muM/liter concentrations) into macromolecular cellular compounds were characterized by multiple K(m) values which were a function of substrate concentration and temperature. It appeared possible that these multiple K(m) values reflected the changing participation of the Embden-Meyerhof-Parnas pathway in glucose metabolism. These results may provide a physiological explanation for the selective enrichment of psychrotrophic freshwater cytophagas. Moreover, they exhibit the limits of interpreting kinetic data based on conventional heterotrophic potential measurements, especially when some complications may arise from temperature and substrate adaptations of the more versatile members of the chemoorganotrophic microflora such as C. johnsonae. PMID- 16346147 TI - Purification and Properties of beta-Glucosidase from Aspergillus terreus. AB - A beta-glucosidase (EC 3.2.1.21) from the fungus Aspergillus terreus was purified to homogeneity as indicated by disc acrylamide gel electrophoresis. Optimal activity was observed at pH 4.8 and 50 degrees C. The beta-glucosidase had K(m) values of 0.78 and 0.40 mM for p-nitrophenyl-beta-d-glucopyranoside and cellobiose, respectively. Glucose was a competitive inhibitor, with a K(i) of 3.5 mM when p-nitrophenyl-beta-d-glucopyranoside was used as the substrate. The specific activity of the enzyme was found to be 210 IU and 215 U per mg of protein on p-nitrophenyl-beta-d-glucopyranoside and cellobiose substrates, respectively. Cations, proteases, and enzyme inhibitors had little or no effect on the enzyme activity. The beta-glucosidase was found to be a glycoprotein containing 65% carbohydrate by weight. It had a Stokes radius of 5.9 nm and an approximate molecular weight of 275,000. The affinity and specific activity that the isolated beta-glucosidase exhibited for cellobiose compared favorably with the values obtained for beta-glucosidases from other organisms being studied for use in industrial cellulose saccharification. PMID- 16346148 TI - Glucose metabolism in sediments of a eutrophic lake: tracer analysis of uptake and product formation. AB - The uptake of glucose and the formation of end products from glucose catabolism have been measured for sediments of eutrophic Wintergreen Lake with a combination of tritiated and C-labeled tracers. Time course analyses of the loss of [H]glucose from sediments were used to establish rate constants for glucose uptake at natural substrate concentrations. Turnover times from these analyses were about 1 min for littoral and profundal sediments. No seasonal or site differences were noted in turnover times. Time course analyses of [U-C]glucose uptake and C-labeled end product formation indicated that glucose mass flow could not be calculated from end product formation since the specific activity of added [C]glucose was significantly diluted by pools of intracellular glucose and glucose metabolites. Mass flow could only be accurately estimated by use of rates of uptake from tracer studies. Intermediate fermentation end products included acetate (71%), propionate (15%), lactate (9%), and only minor amounts of butyrates or valerates. Addition of H(2) to sediments resulted in greater production of lactate (28%) and decreased formation of acetate (50%), but did not affect glucose turnover. Depth profiles of glucose uptake indicated that rates of uptake decreased with depth over the 0- to 18-cm interval and that glucose uptake accounted for 30 to 40% of methanogenesis in profundal sediments. PMID- 16346149 TI - Acetone and Butanol Production by Clostridium acetobutylicum in a Synthetic Medium. AB - The effect of the component concentrations of a synthetic medium on acetone and butanol fermentation by Clostridium acetobutylicum ATCC 824 was investigated. Cell growth was dependent on the presence of Mg, Fe, and K in the medium. Mg and Mn had deleterious effects when in excess. Ammonium acetate in excess caused acid fermentation. The metabolism was composed of two phases: an acid phase and a solvent one. Low concentrations of glucose allowed the first phase only. The theoretical ratio of the conversion of glucose to solvents, which was 28 to 33%, was obtained with the following medium: MgSO(4), 50 to 200 mg/liter; MnSO(4), 0 to 20 mg/liter; KCl, 0.015 to 8 g/liter (an equivalent concentration of K was supplied in the form of KH(2)PO(4) and K(2)HPO(4)); FeSO(4), 1 to 50 mg/liter; ammonium acetate, 1.1 to 2.2 g/liter; para-aminobenzoic acid, 1 mg/liter; biotin, 0.01 mg/liter; glucose, 20 to 60 g/liter. PMID- 16346150 TI - Ethanol Inhibition Kinetics of Kluyveromyces marxianus Grown on Jerusalem Artichoke Juice. AB - The kinetics of ethanol inhibition on cell growth and ethanol production by Kluyveromyces marxianus UCD (FST) 55-82 were studied during batch growth. The liquid medium contained 10% (wt/vol) inulin-type sugars derived from an extract of Jerusalem artichoke (Helianthus tuberosus) tubers, supplemented with small amounts of Tween 80, oleic acid, and corn steep liquor. Initial ethanol concentrations ranging from 0 to 80 g/liter in the liquid medium were used to study the inhibitory effect of ethanol on the following parameters: maximum specific growth rate (mu(max)), cell and ethanol yields, and sugar utilization. It was found that as the initial ethanol concentration increased from 0 to 80 g/liter, and maximum specific growth rate of K. marxianus cells decreased from 0.42 to 0.09 h, whereas the ethanol and cell yields and sugar utilization remained almost constant. A simple kinetic model was used to correlate the mu(max) results and the rates of cell and ethanol production, and the appropriate constants were evaluated. PMID- 16346151 TI - Exopolysaccharide Distribution of and Bioemulsifier Production by Acinetobacter calcoaceticus BD4 and BD413. AB - The heavily encapsulated Acinetobacter calcoaceticus BD4 and the "miniencapsulated" single-step mutant A. calcoaceticus BD413 produced extracellular polysaccharides in addition to the capsular material. The molar ratio of rhamnose to glucose (3:1) in the extracellular BD413 polysaccharide fraction was similar to the composition of the capsular material. In both strains, the increase in capsular polysaccharide was parallel to cell growth and remained constant in stationary phase. The extracellular polysaccharides were detected starting from mid-logarithmic phase and continued to accumulate in the growth medium for 5 to 8 h after the onset of stationary phase. Strain BD413 produced one-fourth the total rhamnose exopolysaccharide per cell that strain BD4 did. Depending on the growth medium, 32 to 63% of the rhamnose polysaccharide produced by strain BD413 was extracellular, whereas in strain BD4 only 7 to 14% was extracellular. In all cases, strain BD413 produced more extracellular rhamnose polysaccharide than strain BD4 did. In glucose medium, strain BD413 also produced approximately 10 times more extracellular emulsifying activity than strain BD4 did. The isolated capsular polysaccharide obtained after shearing of BD4 cells showed no emulsifying activity. Thus, strain BD413 either produces a modified extracellular polysaccharide or excretes an additional substance(s) that is responsible for the emulsifying activity. Emulsions induced by the ammonium sulfate-precipitated BD413 extracellular emulsifier require the presence of magnesium ion and a mixture of an aliphatic and an aromatic hydrocarbon. PMID- 16346152 TI - Nitrous oxide production by organisms other than nitrifiers or denitrifiers. AB - Heterotrophic bacteria, yeasts, fungi, plants, and animal breath were investigated as possible sources of N(2)O. Microbes found to produce N(2)O from NO(3) but not consume it were: (i) all of the nitrate-respiring bacteria examined, including strains of Escherichia, Serratia, Klebsiella, Enterobacter, Erwinia, and Bacillus; (ii) one of the assimilatory nitrate-reducing bacteria examined, Azotobacter vinelandii, but not Azotobacter macrocytogenes or Acinetobacter sp.; and (iii) some but not all of the assimilatory nitrate reducing yeasts and fungi, including strains of Hansenula, Rhodotorula, Aspergillus, Alternaria, and Fusarium. The NO(3)-reducing obligate anaerobe Clostridium KDHS2 did not produce N(2)O. Production of N(2)O occurred only in stationary phase. The nitrate-respiring bacteria produced much more N(2)O than the other organisms, with yields of N(2)O ranging from 3 to 36% of 3.5 mM NO(3). Production of N(2)O was apparently not regulated by ammonium and was not restricted to aerobic or anaerobic conditions. Plants do not appear to produce N(2)O, although N(2)O was found to arise from some damaged plant tops, probably due to microbial growth. Concentrations of N(2)O above the ambient level in the atmosphere were found in human breath and appeared to increase after a meal of high-nitrate food. PMID- 16346153 TI - Dependence of reproduction rate on pressure as a hallmark of deep-sea bacteria. AB - Strains of bacteria in axenic culture were isolated from samples of depths between 1,957 and 10,476 m of the Pacific Ocean. All of the bacteria from this range of depths were barophilic. The pressure at which the rate of reproduction was maximal was found to be correlated with the depth of origin of the isolates. PMID- 16346154 TI - Kinetics of hydrogen consumption by rumen fluid, anaerobic digestor sludge, and sediment. AB - Michaelis-Menten kinetic parameters for H(2) consumption by three methanogenic habitats were determined from progress curve and initial velocity experiments. The influences of mass transfer resistance, endogenous H(2) production, and growth on apparent parameter estimates were also investigated. Kinetic parameters could not be determined for undiluted rumen fluid and some digestor sludge from gas-phase measurements of H(2), since mass transfer of H(2) across the gas-liquid interface was rate limiting. However, accurate values were obtained once the samples were diluted. H(2) consumption by digestor sludge with a long retention time and by hypereutrophic lake sediment was not phase transfer limited. The K(m) values for H(2) uptake by these habitats were similar, with means of 5.8, 6.0, and 7.1 muM for rumen fluid, digestor sludge, and sediment, respectively. V(max) estimates suggested a ratio of activity of approximately 100 (rumen fluid):10 (sludge):1 (sediment); their ranges were as follows: rumen fluid, 14 to 28 mM h; Holt sludge, 0.7 to 4.3 mM h; and Wintergreen sediment, 0.13 to 0.49 mM h. The principles of phase transfer limitation, studied here for H(2), are the same for all gaseous substrates and products. The limitations and errors associated with gas phase determination of kinetic parameters were evaluated with a mathematical model that combined mass transport and Michaelis-Menten kinetics. Three criteria are described which can be used to evaluate the possibility that a phase transfer limitation exists. If it does not exist, (i) substrate consumption curves are Michaelis-Menten and not first order, (ii) the K(m) is independent of initial substrate concentration, and (iii) the K(m) is independent of biomass (V(max)) and remains constant with dilution of sample. Errors in the Michaelis-Menten kinetic parameters are caused by endogenously produced H(2), but they were <15% for rumen fluid and 10% for lake sediment and digestor sludge. Increases in V(max) during the course of progress curve experiments were not great enough to produce systematic deviations from Michaelis-Menten kinetics. PMID- 16346155 TI - Nitrite and nitric oxide as inhibitors of nitrogenase from soybean bacteroids. AB - Nitrite was able to strongly inhibit C(2)H(2) reduction by nitrogenase from soybean bacteroids, whereas H(2) evolution was unaffected under the same conditions. NO inhibited both C(2)H(2) reduction and H(2) evolution; during C(2)H(2) reduction, sensitivity of nitrogenase to NO was higher than to NO(2), and the K(i) values were, respectively, 0.056 and 0.52 mM. Production of NO resulting from a reduction of NO(2) by dithionite in nitrogenase incubations was observed. However, the characteristics of inhibitions and the low level of NO generated by nitrite reduction ruled out the suggestion concerning a direct role of NO to explain the inhibitory effect of NO(2) on nitrogenase. PMID- 16346156 TI - Methane production by fermentor cultures acclimated to waste from cattle fed monensin, lasalocid, salinomycin, or avoparcin. AB - The ability of microorganisms to ferment waste from cattle fed monensin, lasalocid, or salinomycin to methane was determined. Continuously mixed anaerobic fermentors with 3-liter working volumes at 55 degrees C were used; fermentors were fed once per day. Initially, all fermentors were fed waste without antibiotics at 6% volatile solids (VSs, organic matter) and a 20-day retention time (RT) for 60 days. Waste from animals fed monensin, lasalocid, or salinomycin at 29, 20, and 16.5 mg per kg of feed, respectively, was added to duplicate fermentors at the above VSs, and RT. Avoparcin (5 to 45 mg/liter) was not fed to animals but was added directly to duplicate fermentors. Lasalocid and salinomycin had minimal effects on the rate of methane production at RTs of 20 days and later at 6.5 days. Avoparcin caused an increase in organic acids from 599 to 1,672 mg/liter (as acetate) after 4 weeks, but by 6 weeks, acid concentrations declined and the rate of methane production was similar to controls at a 6.5-day RT. The monensin fermentors stopped producing methane 3 weeks after antibiotic addition. However, after a 6-month acclimation period, the microorganisms apparently adapted, and methane production rates of 1.65 and 2.51 liters per liter of fermentor volume per day were obtained with 6% VSs, and RTs of 10 and 6.5 days, respectively. This compares with 1.78 and 2.62 liters/liter per day for controls (P > 0.05). All fermentors that were fed waste containing antibiotics had lower pH values and ammonia and alkalinity concentrations, suggesting less buffering capacity and protein catabolism than in controls. Acclimation results obtained with fermentors at 35 degrees C were similar to those for fermentors at 55 degrees C. These studies indicate that waste from cattle fed these selected growth-promoting antibiotics can be thermophilically fermented to methane at RTs of 6.5 days or longer and VS concentrations of 6%, at rates comparable to waste without antibiotics. PMID- 16346157 TI - Ultrastructural Analysis of Spores and Parasporal Crystals Formed by Bacillus sphaericus 2297. AB - Bacillus sphaericus 2297, growing from a boiled, relatively nontoxic spore inoculum, increased about 30-fold in toxicity for mosquito larvae during early exponential growth but showed an approximately 1,000-fold toxicity increase during the late-exponential phase, as spores began to appear in the culture. The development of spores in the bacterial cells was accompanied by the formation of parasporal crystals. These parasporal crystals appeared during stage III as the forespore septum engulfed the incipient forespore. The paraspores were separated from the forespores by a branch of the exosporium across the cell. Measurements of the parasporal substructure revealed a 6.3-nm distance between the striations. When spores and paraspores were fed to mosquito larvae and the larvae were fixed 15 min after feeding, it was found that the spores remained relatively unchanged but that the matrix of the paraspores was dissolved. After dissolution of the paraspore matrix, a meshlike envelope remained which retained the paraspore shape and which was often in contact with the cross-cell portion of the exosporium. The parasporal crystals may be a source of the mosquito larval toxin in this strain of B. sphaericus, but proof will require their isolation from other cellular components. PMID- 16346159 TI - Measurement of aquatic biodegradation rates by determining heterotrophic uptake of radiolabeled pollutants. AB - [This corrects the article on p. 164 in vol. 44.]. PMID- 16346158 TI - Pock Formation of Streptomyces endus with Production of Phage Taillike Particles. AB - [This corrects the article on p. 1182 in vol. 43.]. PMID- 16346160 TI - Uptake of Branched-Chain Amino Acids by Streptococcus thermophilus. AB - The transport of branched-chain amino acids in Streptococcus thermophilus was energy dependent. The metabolic inhibitors of glycolysis and ATPase enzymes were active, but the proton-conducting uncouplers were not. Transport was optimal at temperatures of between 30 and 45 degrees C and at pH 7.0 for the three amino acids leucine, valine, and isoleucine; a second peak existed at pH 5.0 with valine and isoleucine. By competition and kinetics studies, the branched-chain amino acids were found to share at least a common transport system. PMID- 16346161 TI - Utilization of enzymatically hydrolyzed wood hemicelluloses by microorganisms for production of liquid fuels. AB - Hemicellulose-derived sugars were obtained from a variety of pretreated wood substrates such as water-soluble fractions from steam-exploded aspen, solvent extracted aspen, and commercial xylan. These fractions were enzymatically hydrolyzed by commercial enzyme preparations and by the culture filtrates of eight highly cellulolytic fungi. The sugars released were assayed by high pressure liquid chromatography. Over 30% of the hemicellulose fractions, at a 10% substrate concentration, could be hydrolyzed to monosaccharides. These hemicellulose hydrolysates were used as the substrates for growth of Clostridium acetobutylicum and Klebsiella pneumoniae. Comparatively low butanol values were obtained with C. acetobutylicum, although over 50% of the hemicellulose fraction, at a 1% substrate concentration, could be converted to 2,3-butanediol, ethanol, and acetic acid by K. pneumoniae. PMID- 16346162 TI - Methanogenesis from Choline by a Coculture of Desulfovibrio sp. and Methanosarcina barkeri. AB - A sulfate-reducing vibrio was isolated from a methanogenic enrichment with choline as the sole added organic substrate. This organism was identified as a member of the genus Desulfovibrio and was designated Desulfovibrio strain G1. In a defined medium devoid of sulfate, a pure culture of Desulfovibrio strain G1 fermented choline to trimethylamine, acetate, and ethanol. In the presence of sulfate, more acetate and less ethanol were formed from choline than in the absence of sulfate. When grown in a medium containing sulfate, a coculture of Desulfovibrio strain G1 and Methanosarcina barkeri strain Fusaro degraded choline almost completely to methane, ammonia, and hydrogen sulfide and presumably to carbon dioxide. Methanogenesis occurred in two distinct phases separated by a lag of about 6 days. During the first phase of methanogenesis choline was completely converted to trimethylamine, acetate, hydrogen sulfide, and traces of ethanol by the desulfovibrio. M. barkeri fermented trimethylamine to methane, ammonia, and presumably carbon dioxide via dimethyl- and methylamine as intermediates. Simultaneously, about 60% of the acetate expected was metabolized. In the second phase of methanogenesis, the residual acetate was almost completely catabolized. PMID- 16346163 TI - Growth Kinetics and Yield Coefficients of the Extreme Thermophile Thermothrix thiopara in Continuous Culture. AB - Thermothrix thiopara did not appear to be stressed at high temperature (72 degrees C). Both the actual and theoretical yields were higher than those of analogous mesophilic sulfur bacteria, and the specific growth rate (mu(max)) was more rapid than that of most autotrophs. The specific growth rate (0.58 h), specific maintenance rate (0.11 h), actual molar growth yield at mu(max) (Y(max) = 16 g mol), and theoretical molar growth yield (Y(G) = 24 g mol) were all higher for T. thiopara (72 degrees C) than for mesophilic (25 to 30 degrees C) Thiobacillus spp. The growth efficiencies for T. thiopara at 70 and 75 degrees C (0.84 and 0.78) were significantly higher than at 65 degrees C (0.47). Corresponding specific maintenance rates were highest at 65 degrees C (0.41 h) and lowest at 70 and 75 degrees C (0.11 and 0.15 h, respectively). Growth efficiencies of metabolically similar mesophiles were generally higher than for T. thiopara. However, the actual yields at mu(max) were higher for T. thiopara because its theoretical yield was higher. Thus, at 70 degrees C, T. thiopara was capable of deriving more metabolically useful energy from thiosulfate than were mesophilic sulfur bacteria at 25 and 30 degrees C. The low growth efficiency of T. thiopara reflected higher maintenance expenditures. T. thiopara had higher maintenance rates than Thiobacillus ferroxidans or Thiobacillus denitrificans, but also attained higher molar growth yields. It is concluded that sulfur metabolism may be more efficient overall at extremely high temperatures due to increased theoretical yields despite increased maintenance requirements. PMID- 16346164 TI - Sulfate reducers can outcompete methanogens at freshwater sulfate concentrations. AB - Acetate and hydrogen metabolism by sulfate reducers and methanogens in the profundal sediments of an oligotrophic lake were examined. Inhibition of sulfate reduction with molybdate stimulated methane production from both hydrogen and acetate. Molybdate did not stimulate methane production in sediments that were preincubated to deplete the sulfate pool. Sulfate reduction accounted for 30 to 81% of the total of terminal metabolism proceeding through sulfate reduction and methane production in Eckman grab samples of surface sediments. The ability of sulfate reducers to effectively compete with methanogens for acetate was related to the sulfate reducers' lower half-saturation constant for acetate metabolism at in situ sulfate concentrations. Processes other than sulfate reduction and methanogenesis consumed hydrogen at elevated hydrogen partial pressures and prevented a kinetic analysis of hydrogen uptake by sulfate reducers and methanogens. The demonstration that sulfate reducers can successfully compete with methanogens for hydrogen and acetate in sediments at in situ sulfate concentrations of 60 to 105 muM extends the known range of sediment habitats in which sulfate reduction can be a dominant terminal process. PMID- 16346165 TI - Substrates for sulfate reduction and methane production in intertidal sediments. AB - The activity of and potential substrates for methane-producing bacteria and sulfate-reducing bacteria were examined in marsh, estuary, and beach intertidal sediments. Slow rates of methane production were detected in all sediments, although rates of sulfate reduction were 100- to 1,000-fold higher. After sulfate was depleted in sediments, the rates of methane production sharply increased. The addition of methylamine stimulated methanogenesis in the presence of sulfate, and [C]methylamine was rapidly converted to CH(4) and CO(2) in freshly collected marsh sediment. Acetate, hydrogen, or methionine additions did not stimulate methanogenesis. [methyl-C]methionine and [2-C]acetate were converted to CO(2) and not to CH(4) in fresh sediment. No reduction of CO(2) to CH(4) occurred in fresh sediment. Molybdate, an inhibitor of sulfate reduction, inhibited [2-C]acetate metabolism by 98.5%. Fluoracetate, an inhibitor of acetate metabolism, inhibited sulfate reduction by 61%. These results suggest that acetate is a major electron donor for sulfate reduction in marine sediments. In the presence of high concentrations of sulfate, methane may be derived from novel substrates such as methylamine. PMID- 16346166 TI - Potential of lactic streptococci to produce bacteriocin. AB - A survey was made on the bacteriocin-producing potential of lactic streptococci. Bacteriocin-like activities were isolated and partially purified from about 5% of the 280 strains investigated. The frequency of production varied from about 1% in Streptococcus lactis subsp. diacetylactis to 9 and 7.5% in S. lactis and Streptococcus cremoris, respectively. Eight strains of S. cremoris produced bacteriocins which, on the basis of heat stability at different pH values and inhibitory spectrum, could be divided into four types. From 54 S. lactis strains, 5 strains produced inhibitory substances, namely, three nisin-like antibiotics and two different bacteriocins. Only 1 of 93 S. lactis subsp. diacetylactis strains produced a bacteriocin which was very similar to bacteriocins of type I in S. cremoris. All of the bacteriocins that were partially purified by ammonium sulfate precipitation showed very limited inhibitory spectra. Most of the lactic streptococci and a few members of the genera Clostridium, Leuconostoc, and Pediococcus were inhibited. None of the bacteriocins acted on gram-negative bacteria. The bacteriocinogenic strains were also characterized on the basis of plasmid content. All strains possessed between one and nine plasmids ranging from 1 to 50 megadaltons. PMID- 16346167 TI - Breakdown of diazotized proteins and synthetic substrates by rumen bacterial proteases. AB - Several different kinds of substrate were used to investigate the proteolytic activity of rumen bacteria and of proteases released from rumen bacteria by blending ("coat proteases"). These substrates included diazotized feed proteins and diazotized soluble and insoluble pure proteins. It was concluded that, while solubility was an important factor, the secondary and tertiary structure of a protein had a major influence on its rate of digestion. The resistance of elastin congo red to digestion indicated that similar fibrous proteins in plant material might resist proteolytic attack by rumen bacteria. Coat proteases had a broad specificity, including several exo- and endopeptidase activities, as determined by using synthetic peptide substrates. PMID- 16346168 TI - Halophilic Nuclease of a Moderately Halophilic Bacillus sp.: Production, Purification, and Characterization. AB - A moderately halophilic bacterium, Bacillus sp., isolated from rotting wood on the seashore in Nauru, produced an extracellular nuclease when cultivated aerobically in media containing 1 to 2 M NaCl. The enzyme was purified from the culture filtrate to an electrophoretically homogeneous state by ethanol precipitation, DEAE-Sephadex A-50 column chromatography, and Sephadex G-200 gel filtration. The enzyme consisted of two charge isomers and showed both RNase and DNase activities. Molecular weight was estimated to be 138,000 by Sephadex G-200 gel filtration. The enzyme had marked halophilic properties, showing maximal activities in the presence of 1.4 to 3.2 M NaCl or 2.3 to 3.2 M KCl. The enzyme hydrolyzed thymidine-5'-monophosphate-p-nitrophenyl ester at a rate that increased with NaCl concentration up to 4.8 M. In the presence of both Mg and Ca, activity was greatly enhanced. The activity was lost by dialysis against water and low-salt buffer, but it was protected when 10 mM Ca was added to the dialysis buffer. When the inactivated enzyme was dialyzed against 3.5 M NaCl buffer as much as 68% of the initial activity could be restored. The enzyme exhibited maximal activity at pH 8.5 and at 50 degrees C on DNA and at 60 degrees C on RNA and attacked RNA and DNA exonucleolytically and successively, producing 5' mononucleotides. PMID- 16346169 TI - Method for establishing a bacterial inoculum on corn roots. AB - Few bacteria from the corn rhizosphere grew in media with 50 mug of mancozeb per ml. A mancozeb-resistant Pseudomonas strain from the rhizosphere was serially subcultured in media containing mancozeb and spectinomycin until it was resistant to 175 mug of mancozeb and 850 mug of spectinomycin per ml. The population of the pseudomonad added to soil fell to low numbers in 6 days in unamended or glucose amended soil, but its numbers exceeded 10/g for at least 12 days if the soil was supplemented with mancozeb. The numbers of this organism remained small on corn roots derived from untreated, inoculated seeds, but the population was two or more orders of magnitude greater on roots derived from mancozeb-coated seeds. The abundance of the inoculum strain on the 3-cm portion of roots nearest the stem declined markedly after about 1 week, but applying urea to the foliage reduced or prevented the decline. The numbers of the pseudomonad on segments of roots 3- to 6- and 6- to 9-cm from the stem were higher on plants derived from the mancozeb coated seeds. Applying spectinomycin to the foliage did not promote growth of the bacterium. This method is proposed as a means to establish an introduced bacterium on plant roots. PMID- 16346170 TI - Improvements in and environmental applications of double-vial radiorespirometry for the study of microbial mineralization. AB - Several variations in the scintillation mixture and the filter paper arrangements for double-vial radiorespirometry were compared. Improved efficiencies (44%) and shorter response times were found by adding wetting agents and methanolic NaOH to the scintillation mixture in the filter paper. The scintillation chemicals used did not contain dioxane and were found to be nontoxic to the test microbiota in this system. Covering the inner reaction vial with aluminum foil minimized the reduction in counting efficiency when testing colored or dense environmental samples. Mineralization rates were determined with C-labeled glucose, acetate, and glutamate and [C]cellulose- and [C]lignin-labeled lignocellulose for composting cow manure, forest soil, and arctic lake sediment microbiota. This improved method can be used in a variety of procedures involving the measurement of microbial mineralizations of organic compounds. Since no liquid scintillation cocktail is used for counting, the radioactive wastes are aqueous or can be incinerated, making disposal easy. PMID- 16346171 TI - Isolation and characterization of an h(2)-oxidizing thermophilic methanogen. AB - A thermophilic methanogen was isolated from enrichment cultures originally inoculated with sludge from an anaerobic kelp digester (55 degrees C). This isolate exhibited a temperature optimum of 55 to 60 degrees C and a maximum near 70 degrees C. Growth occurred throughout the pH range of 5.5 to 9.0, with optimal growth near pH 7.2. Although 4% salt was present in the isolation medium, salt was not required for optimal growth. The thermophile utilized formate or H(2) CO(2) but not acetate, methanol, or methylamines for growth and methanogenesis. Growth in complex medium was very rapid, and a minimum doubling time of 1.8 h was recorded in media supplemented with rumen fluid. Growth in defined media required the addition of acetate and an unknown factor(s) from digester supernatant, rumen fluid, or Trypticase. Cells in liquid culture were oval to coccoid, 0.7 to 1.8 mum in diameter, often occurring in pairs. The cells were easily lysed upon exposure to oxygen or 0.08 mg of sodium dodecyl sulfate per ml. The isolate was sensitive to tetracycline and chloramphenicol but not penicillin G or cycloserine. The DNA base composition was 59.69 mol% guanine plus cytosine. PMID- 16346172 TI - Dialysis membrane technique for studying microbial interaction. AB - A dialysis membrane method is described which allows (i) cultivation of fungi on an agar support, (ii) observation of growth and development by direct light microscopy, (iii) transfer of cultures from agar surfaces for subsequent treatments or for biochemical analysis, and (iv) preparation for scanning electron microscopy. The method is used routinely in studies of fungus-nematode and fungus-fungus interactions. PMID- 16346173 TI - Tubular heads in bacteriophages from lactic streptococci. AB - Tubular bacteriophage heads were observed in the lysate of two phages from Streptococcus lactis obtained from single plaques without mutagenesis. The frequency of appearance of the tubular heads was 2.5 and 16%. PMID- 16346174 TI - Transformation of Clostridium acetobutylicum Protoplasts with Bacteriophage DNA. AB - Techniques for the transformation of Clostridium acetobutylicum protoplasts with bacteriophage DNA are described. Transformation required regeneration of protoplasts and a 2-h eclipse period. PMID- 16346175 TI - Azo Dye Colony Staining Technique To Detect Fungal Mutants Lacking beta Glucosidase Activity. AB - A simple, rapid, convenient, easily interpreted azo dye colony-staining technique was developed to detect fungal mutants lacking beta-glucosidase activity on a 6 bromo-2-naphthyl substrate. The procedure includes net replication of colonies and chemical restriction of colony growth to allow testing of many colonies per culture dish. PMID- 16346176 TI - Prototheca zopfii Kruger Strain UMK-13 Growth on Acetate or n-Alkanes. AB - A new strain of Prototheca zopfii Kruger was grown on acetate or on pure n alkanes. A maximum acetate-supported exponential growth of 12 divisions day occurred at pH 5 and 30 degrees C. At 25 degrees C, growth on n-alkanes was almost as fast, but no growth occurred at 30 degrees C. After 4 days at 25 degrees C, 34 to 45% of the n-alkanes had been removed, whereas at 21 degrees C and slower growth, utilization was twofold greater after 15 days. Rates of growth and utilization increased markedly after a point of sudden emulsification. PMID- 16346177 TI - Congo Red Absorption by Rhizobium leguminosarum. AB - Congo red absorption is generally considered a contraindication of Rhizobium. However, R. leguminosarum takes up the dye on yeast extract-mannitol agar. The uptake of congo red varies among strains of R. leguminosarum, as shown elsewhere with strains of R. trifolii and R. meliloti. Congo red absorption does not distinguish rhizobia from other bacteria, but may be useful as a strain marker. PMID- 16346178 TI - Production of rosamicin: improvement of synthetic medium. AB - Rosamicin is one of the important macrolide antibiotics that has clinical efficacy and broad-spectrum antibacterial activity. Using a mutant strain of Micromonospora rosaria (NRRL 3718), a chemically defined medium was developed, and some fermentation conditions that are important to rosamicin biosynthesis were optimized to achieve rosamicin productivity of 230 mug/ml. Soluble starch and l-asparagine were found to be the best carbon and nitrogen sources, and a stimulative effect of magnesium and zinc ions was also found. The medium developed contains: soluble starch, 4%; l-asparagine, 0.15%; K(2)HPO(4), 0.075%; CaCO(3), 0.6%; MgSO(4) . 7H(2)O, 0.05%; FeSO(4) . 7H(2)O, 10 M; CuSO(4) . 5H(2)O, 10 M; ZnSO(4) . 7H(2)O, 10 M; and MnSO(4) . (4-6)H(2)O, 10 M. The required air supply was about 40 mmol of O(2) liter . h . atm, and the favorable culture temperature was 28 to 29 degrees C. PMID- 16346179 TI - Multiplication of Legionella pneumophila in Unsterilized Tap Water. AB - [This corrects the article on p. 1333 in vol. 43.]. PMID- 16346180 TI - Responses of marine bacteria under starvation conditions at a solid-water interface. AB - Size changes during starvation of 17 marine bacterial isolates at a solid-water interface and in the liquid phase were examined. Twelve rod-shaped, hydrophilic bacteria decreased in size more rapidly at the solid surface than in the liquid phase, a result parallel to that observed previously for one of the strains at an air-water interface. On the other hand, three rod-shaped, hydrophobic bacteria diminished in size more rapidly in the liquid phase than at the solid-water interface. The rapid size decrease (defined here as the dwarfing phase) in either situation appeared to be an active process which occurred more rapidly when the cells were in an early stage of logarithmic growth at the onset of starvation. Dwarfing was reversibly inhibited by low temperature and low pH but was not inhibited by chloramphenicol. Three coccoidal bacteria showed little tendency to become smaller upon starvation in the liquid phase or at a surface. PMID- 16346181 TI - Spatial distribution of biochemical parameters indicating biomass and community composition of microbial assemblies in estuarine mud flat sediments. AB - The spatial distribution of communities was examined in estuarine mud flat sediments by the biochemical analysis of the lipids and lipid components extracted from the sediments. Total phospholipid was used as a measure of total biomass, and fatty acids were used as indicators of community composition. Comparisons were made among 2- by 2-m (location) and 0.2- by 0.2-m (cluster) sampling plots by using a nested analysis of variance to design an optimal sampling strategy to define the microbial content of a large, relatively homogenous area. At two of the three stations, a 2- by 2-m plot was representative of the station, but 0.2- by 0.2-m areas were in no case representative of the station. The biomass measured by the extractable phospholipid and the total lipid palmitic acid showed excellent correlation with the fatty acid "signatures" characteristic of bacteria, but showed a lower correlation with the long-chain polyenoic fatty acids characteristic of the microfauna. PMID- 16346182 TI - Linalyl Acetate Is Metabolized by Pseudomonas incognita with the Acetoxy Group Intact. AB - Metabolism of linalyl acetate by Pseudomonas incognita isolated by enrichment culture on the acyclic monoterpene alcohol linalool was studied. Biodegradation of linalyl acetate by this strain resulted in the formation of linalool, linalool 8-carboxylic acid, oleuropeic acid, and Delta-4-acetoxy-4-methyl hexenoic acid. Cells adapted to linalyl acetate metabolized linalyl acetate-8-aldehyde to linalool-8-carboxylic acid, linalyl acetate-8-carboxylic acid, Delta-4-acetoxy-4 methyl hexenoic acid, and geraniol-8-carboxylic acid. Resting cell suspensions previously grown with linalyl acetate oxidized linalyl acetate-8-aldehyde to linalyl acetate-8-carboxylic acid, Delta-4-acetoxy-4-methyl hexenoic acid, and pyruvic acid. The crude cell-free extract (10,000 g of supernatant), obtained from the sonicate of linalyl acetate-grown cells, was shown to contain enzyme systems responsible for the formation of linalyl acetate-8-carboxylic acid and linalool-8-carboxylic acid from linalyl acetate. The same supernatant contained NAD-linked alcohol and aldehyde dehydrogenases involved in the formation of linalyl acetate-8-aldehyde and linalyl acetate-8-carboxylic acid, respectively. On the basis of various metabolites isolated from the culture medium, resting cell experiments, growth and manometric studies carried out with the isolated metabolites as well as related synthetic analogs, and the preliminary enzymatic studies performed with the cell-free extract, a probable pathway for the microbial degradation of linalyl acetate with the acetoxy group intact is suggested. PMID- 16346183 TI - Relationship Between Physiological Status and Formation of Extracellular Polysaccharide Glycocalyx in Pseudomonas atlantica. AB - Marine pseudomonads, such as Pseudomonas atlantica, are readily isolated from sediments. These organisms form extracellular polysaccharide polymers (glycocalyx). The factors affecting the composition and amount of glycocalyx in batch culture of these organisms were examined. The formation of glycocalyx was stimulated by the inclusion of galactose as the carbon source and by increased surface area resulting from addition of sand to the medium. The composition of the glycocalyx changed during the growth cycle, with a marked increase in the proportions and absolute amounts of uronic acids as the rate of synthesis increased. In estuarine sediments, the glycocalyx contained a carbon content at least as great as in the microbes themselves. The greatest accumulation of these polymers occurred late in the stationary phase when the physiological status of the cells, as measured by the adenylate energy charge, showed maximal stress. Maximal formation of glycocalyx possibly could be used as an estimate of the nutritional status of these microbes. PMID- 16346184 TI - Polymeric Beta-Hydroxyalkanoates from Environmental Samples and Bacillus megaterium. AB - The procaryotic endogenous storage polymer known as poly-beta-hydroxybutyrate is actually a mixed polymer of short-chain beta-hydroxy fatty acids. A method for the quantitative recovery of this mixed polymer, called poly-beta hydroxyalkanoate (PHA), with analysis by capillary gas-liquid chromatography, showed the presence of at least 11 short-chain beta-hydroxy acids in polymers extracted from marine sediments. Polymers extracted from Bacillus megaterium monocultures were also a complex mixture of beta-hydroxy acids with chain lengths between four and eight carbons. Lyophilized sediments were extracted in a modified Soxhlet extractor, and the polymer was purified with ethanol and diethyl ether washes. The purified polymer was treated with ethanol-chloroform hydrochloric acid (8.5:2.5:1) for 4 h at 100 degrees C, a treatment which resulted in the formation of the ethyl esters of the constituent beta-hydroxy acids. Subsequent assay of the products by gas-liquid chromatography indicated excellent reproducibility and sensitivity (detection limit, 100 fmol). Disturbing sediments mechanically or adding natural chelators increased all major PHA components relative to the bacterial biomass. Gardening of sedimentary microbes by Clymenella sp., an annelid worm, induced decreases in PHA, with changes in the relative proportion of component beta-hydroxy acids. The concentration of PHA relative to the bacterial biomass can reflect the recent metabolic status of the microbiota. PMID- 16346185 TI - End Products of Glucose Fermentation by Brochothrix thermosphacta. AB - Anaerobically, Brochothrix thermosphacta fermented glucose primarily to l lactate, acetate, formate, and ethanol. The ratio of these end products varied with growth conditions. Both the presence of acetate and formate and a pH below about 6 increased l-lactate production from glucose. Small amounts of butane-2,3 diol were also produced when the pH of the culture was low ( 1 R-COOH + 1 CO(2) + 1 NH(3) + 2 trimethylamine + 2 acetate. Growth and production of trimethylamine were strictly dependent on the presence of selenite in the medium. With cell suspensions it was shown that C. sporogenes was unable to catabolize betaine as a single substrate. Betaine, however, was reduced to trimethylamine and acetate under an atmosphere of molecular hydrogen. For the reduction of betaine by cell extracts of C. sporogenes, dimercaptans such as 1,4-dithiothreitol could serve as electron donors. No betaine reductase activity was detected in cells grown in a complex medium without betaine. The pH optimum of betaine reductase was at pH 7.3. When C. sporogenes was cocultured with Methanosarcina barkeri strain Fusaro on betaine together with l-alanine, an almost complete conversion of the two substrates to CH(4), NH(3), and presumably CO(2) was observed. PMID- 16346197 TI - Dynamics of Baculovirus Growth and Dispersal in Mamestra brassicae L. (LepidopteraNoctuidae) Larval Populations Introduced into Small Cabbage Plots. AB - The nuclear polyhedrosis virus of Mamestra brassicae has been studied in larval populations of the moth introduced into small plots of cabbages. Primary dispersal of virus from single foci of infected larvae resulted from enhanced movement of the larvae, which colonized new plants logarithmically. Virus growth within the host population was quantified, and infection of young larvae in the following generation was related directly to the concentration of virus produced during the primary phase. Secondary cycling of virus resulted in dispersal of inoculum from multiple foci, and a large proportion of plants were ultimately colonized by infected larvae. The dynamics of virus growth during secondary dispersal were quantified and contrasted with results from the primary phase. The significance of these results is discussed in relation to possible control of insect pests through dispersal of virus by the host insect. PMID- 16346198 TI - Production and consumption of hydrogen in a eutrophic lake. AB - The vertical distribution of hydrogen was measured in the Loclat, a eutrophic and holomictic lake near Neuchatel, Switzerland, before and during summer stratification. H(2) concentrations decreased with depth in the anaerobic hypolimnion and were often below the detection limit (2.5 nl of H(2) liter) in the water adjacent to the lake sediment. H(2) was apparently not released from the lake sediment. The highest H(2) concentrations (>4 mul of H(2) liter) were observed in the aerobic water of the epilimnion and metalimnion. There, the H(2) concentrations changed with time, indicating a turnover of H(2). The H(2) production processes could not be studied in the laboratory since incubation of water samples in light or darkness did not result in H(2) production but rather always in H(2) consumption. The possible role of cyanobacteria and algae for H(2) production is discussed. Aerobic or anaerobic H(2) consumption activities were observed at all depths of the water column, with highest activities in the hypolimnion. Aerobic H(2) consumption activity was insensitive to azide inhibition, but sensitive to heat, mercuric chloride, or cyanide. It was restricted to a particle fraction of 0.2 to 3.0 mum in size, so that it must be due to single bacterial cells. Aerobic hydrogen bacteria, on the other hand, occurred in clusters of >3.0 mum. Therefore, the hydrogen bacteria could not have caused the H(2) consumption in lake water. The aerobic H(2) consumption activity followed Michaelis-Menten kinetics, with a K(m) of 67 nM H(2). This is an exceptionally low value compared with K(m) values of hydrogenases in hydrogen bacteria and other species, but is similar to that for H(2)-decomposing abiontic soil hydrogenases. PMID- 16346199 TI - Biological Control of Olive Green Mold in Agaricus bisporus Cultivation. AB - Successful methods to control the damaging weed mold Chaetomium olivaceum (olive green mold) in mushroom beds are not presently known. An attempt was made to control C. olivaceum by biological means. A thermophilic Bacillus sp. which showed dramatic activity against C. olivaceum on Trypticase soy agar (BBL Microbiology Systems)-0.4% yeast extract agar plates was isolated from commercial mushroom compost (phase I). When inoculated into conventional and hydroponic mushroom beds, the bacillus not only provided a significant degree of protection from C. olivaceum, but also increased yields of Agaricus bisporus. PMID- 16346200 TI - Bacterial activity associated with the decomposition of woody substrates in a stream sediment. AB - Ground bark and heartwood from Alnus rubra and Pseudotsuga menziesii were added to a muddy sediment from a small Oregon stream and incubated in situ. Carbon dioxide and methane production rates were increased by all amendments, the biggest increase being shown with A. rubra wood. Except for sediment amended with A. rubra wood, nitrogen fixation rates from all treatments (including the control) were approximately 0.1 nmol/g per h throughout the 6-month study period. Contrary to expectations, neither bark had a noticeable adverse effect on microbial activity, but the A. rubra wood promoted nitrogen fixation. These results help to explain the faster rate of decomposition of A. rubra wood in water compared with that of P. menziesii described in the literature. The uptake kinetics of glucose (V(max)) did not follow the same pattern as gas evolution. PMID- 16346201 TI - Microbial Transformation of Squalene: Terminal Methyl Group Oxidation by Corynebacterium sp. AB - Corynebacterium sp. strain SY-79 was isolated from soil, using squalene as a carbon source. Microbiological properties of this bacterium are described. The metabolic product of this bacterium from squalene was identified as 2,6,10,15,19, 23-hexamethyl-2,6,10,14,18,22-tetracosahexaenedioic acid (squalenedioic acid). PMID- 16346202 TI - Production of Antimicrobial and Bacteriocin-Like Substances by Rhizobium trifolii. AB - Rhizobium trifolii strains IARI and Rel-1 produced substances with broad and narrow activity spectra, respectively. Reproducible inhibitory zones of various sizes produced by R. trifolii IARI (2 to 14 mm) and R. trifolii Rel-1 (2 to 6 mm) were detected, depending upon the indicator organism used. The maximum production of these substances by both strains of R. trifolii was observed on l-arabinose agar. A preliminary characterization of the antimicrobial substance produced by strain IARI showed resistance to heat (75 to 80 degrees C for 45 min), trypsin, lysozyme, DNase I, and RNase A. On the other hand, the substance produced by strain Rel-1 showed sensitivity to heat (75 to 80 degrees C for 45 min) and trypsin, but resistance to lysozyme, RNase A, and DNase I. PMID- 16346203 TI - Population Changes and Persistence of Rhizobium phaseoli in Soil and Rhizospheres. AB - The impact of legume cultivation on the establishment and persistence of an inoculant strain of Rhizobium phaseoli and its ability to compete with a resident population of R. phaseoli for nodule occupancy was examined utilizing strain specific fluorescent antibodies. The soil (Hubbard loamy sand) was inoculated homogeneously with 5 x 10 cells per g of soil and confined in plastic cylinders kept in field plots. Inoculated and uninoculated cylinders were either left fallow or planted to two seeds of legumes. Two hosts, navy bean (Phaseolus vulgaris L.) cv. Seafarer and snap bean cv. Picker, as well as a nonhost, soybean (Glycine max (L.) Merr.) cv. Wilkin, were used. Inoculant Viking 1 was highly stimulated in all three rhizospheres sampled at 6 (flowering), 10 (podfill), and 17 (decay) weeks and in the following spring, whereas counts in fallow soil decreased rapidly. Although the overwintering population remained highest in the vicinity of decaying host roots, Viking 1 persisted, even in fallow soil, to produce abundant nodulation of host plants the following spring. Viking 1 was an excellent competitor for nodulation sites on the roots of the hosts; it thoroughly outcompeted the resident population of R. phaseoli, occupying virtually 100% of the nodules under inoculated conditions in all experiments. PMID- 16346204 TI - Pseudomonas aeruginosa Chemotaxis Associated with Blooms of N(2)-Fixing Blue Green Algae (Cyanobacteria). AB - Pseudomonas aeruginosa (Schroeter) Migula, a numerically significant bacterium found during N(2)-fixing blooms of the blue-green algae (cyanobacteria) Anabaena sp. in the Chowan River, North Carolina, was chemotactically attracted to amino acids when tested in a radioassay. The bacterium was labeled with P(i), and the disintegrations per minute determined by liquid scintillation counting were proportional to the number of cells accumulating in microcapillaries containing amino acids. Positive chemotaxis was observed toward all of the amino acids tested, although the degrees of response varied. Since many nitrogen-fixing blue green algae secrete nitrogenous compounds, this attraction may be instrumental in establishing a symbiotic relationship between this bacterium and blue-green algae in freshwater. PMID- 16346205 TI - Nitrogenase activity (acetylene reduction) of root-associated, cold-climate azospirillum, enterobacter, Klebsiella, and pseudomonas species during growth on various carbon sources and at various partial pressures of oxygen. AB - A comprehensive view of the diazotrophic bacterial flora of plants requires that attention be paid to the appropriate carbon and oxygen requirements during isolation of the bacteria. Twenty compounds (monosaccharides, disaccharides, polyols, and organic acids) were therefore examined as carbon and energy sources for nitrogenase activity in semisolid stab cultures at pO(2) values of 0.21, 0.02, and 12. Swelling of crystals occurred in the buffer between pH 10 and 11, and crystals dissolved in the same buffer supplemented with gut juice protease of the silkworm Bombyx mori. The proteolytic dissolution of crystals occurred after a time lag of several minutes in 0.1 M carbonate-NaOH buffer, pH 10.2. The time lag was not observed when crystals were suspended in the buffer for 30 min before the addition of protease. After the dissolution of the crystals and further degradation of the solubilized protein, the appearance of a toxic protein with a molecular weight of 59,000, designated P-59, was observed. Lower-molecular-weight peptides (less than 40,000) showed no toxicity to the silkworm larvae on feeding. Digestion of the 120,000-dalton subunit of the crystal by gut juice protease also produced P-59. These observations suggest the occurrence of a similar process in vivo, i.e., the swelling of crystals due to the alkalinity of gut juice and the production of P 59, dependent on the hydrolysis of swollen crystals by gut juice protease. PMID- 16346207 TI - Enzyme-Linked Immunosorbent Assay for Quantitative Detection of Bacillus thuringiensis Crystal Protein. AB - Accurate measurement of the toxic protein crystal produced during deep-tank fermentation of Bacillus thuringiensis is critical for optimum process yield. The currently accepted method is a bioassay that requires more time to generate data than to complete the fermentation itself. A noncompetitive enzyme-linked immunosorbent assay has been developed with purified B. thuringiensis crystals to generate rabbit antiserum. This technique gives a quantitative crystal protein value with a colorimetric endpoint for either liquids or powders within 4 h of sampling. Reproducibility of this enzyme-linked immunosorbent assay satisfies criteria for use in a commercial process. PMID- 16346208 TI - Lipid-Enhanced Ethanol Production by Kluyveromyces fragilis. AB - The fermentation ability of a strain of Kluyveromyces fragilis, already selected for rapid lactose-fermenting capability, was improved dramatically by the addition of unsaturated fatty acids and ergosterol to the medium. The fermentation time of a 20% whey-lactose medium was decreased from over 90 h to less than 60 h. The lipids were shown to be taken up by the organism, and the effects on specific growth rate and biomass production were determined. PMID- 16346209 TI - Microbial production of lysine and threonine from whey permeate. AB - Extracellular accumulation of lysine and threonine was investigated in modified whey permeate by using Brevibacterium lactofermentum ATCC 21086 and Escherichia coli ATCC 21151. Whey permeate was prepared from whey by membrane ultrafiltration, and lactose was hydrolyzed by treating permeate with HCl or beta galactosidase. The highest amount of lysine (3.3 g/liter) was produced from a mixture of acid-hydrolyzed whey permeate and yeast extract (0.2%). The highest amount of threonine (3.6 g/liter) was produced from a mixture of whey permeate, (NH(4))(2)SO(4) (1.4%), yeast extract (0.1%), and Na(2)CO(3) (0.3%). PMID- 16346210 TI - Microbial Transformations of Natural Antitumor Agents: Products of Rotenone and Dihydrorotenone Transformation by Cunninghamella blakesleeana. AB - Various species of Absidia, Aspergillus, Cunninghamella, Trichothecium, Penicillium, and Phanerochaete were found to transform rotenone to one or more metabolites. Two biotransformation products were isolated from a preparative scale incubation of rotenone with Cunninghamella blakesleeana and identified as 1',2'-dihydro-1',2'-dihydroxyrotenone and 3'-hydroxyrotenone (amorphigenin). The catalytic reduction of the isopropylene side chain of rotenone resulted in the formation of 1',2'-dihydrorotenone. The latter was transformed by C. blakesleeana to 2'-hydroxy-1',2'-dihydrorotenone. PMID- 16346211 TI - Application of response surface methodology to evaluation of bioconversion experimental conditions. AB - Using Candida tenuis, a yeast isolated from the digestive tube of the larva of Phoracantha semipunctata (Cerambycidae, Coleoptera), we were able to demonstrate the bioconversion of citronellal to citronellol. Response surface methodology was used to achieve the optimization of the experimental conditions for that bioconversion process. To study the proposed second-order polynomial model, we used a central composite experimental design with multiple linear regression to estimate the model coefficients of the five selected factors believed to influence the bioconversion process. Only four were demonstrated to be predominant: the incubation pH, temperature, time, and the amount of substrate. The best reduction yields (close to 90%) were obtained with alkaline pH conditions (pH 7.5), a low temperature (25 degrees C), a small amount of substrate (15 mul), and short incubation time (16 h). This methodology was very efficient: only 36 experiments were necessary to assess these conditions, and model adequacy was very satisfactory as the coefficient of determination was 0.9411. PMID- 16346212 TI - Tn5-Induced Mutations in the Enterobacterial Phytopathogen Erwinia chrysanthemi. AB - Escherichia coli (2492/pJB4JI) matings with Erwinia chrysanthemi produced kanamycin resistant (Km) transconjugants, a majority of which were gentamicin sensitive (Gm). A small proportion (about 0.8%) of the Km Gm clones were either auxotrophic or failed to catabolize galacturonate (Gtu). The R plasmid (pJB4JI) DNA was detected in the parent E. coli strain and in a Km Gm transconjugant, but not in Km GmE. chrysanthemi strains carrying Tn5-induced mutations. In Hfr crosses, Km (Tn5) was found linked with most mutations. A majority (>95%) of prototrophic recombinants were Km, except for Leu and Arg recombinants which were 30 to 50% Km. Spontaneous revertants were obtained for all markers except car, gtu, lys, thr, and trp. Prototrophic revertants, with the exception of Met, Leu, or His clones, were Km. We conclude from both genetic and physical data that Tn5 transposed from pJB4JI into different sites on the chromosome of E. chrysanthemi. PMID- 16346213 TI - Exogenous Valine Reduces Conversion of Leucine to 3-Methyl-1-Butanol in Saccharomyces cerevisiae. AB - Mutant strains of the yeast Saccharomyces cerevisiae that require branched-chain amino acids must be supplemented with large concentrations (up to 10 mM) of these amino acids to satisfy their nutritional requirement. The utilization of one branched-chain amino acid, leucine, was examined in several leul strains of yeast grown aerobically in a glucose-ammonium salts minimal medium containing a limiting concentration (0.2 mM) of leucine. In this medium, the leucine requirement of the auxotrophic strains could be reduced by valine, another branched-chain amino acid. Increasing the valine concentration increased the cell yields of cultures and also reduced the levels of 3-methyl-1-butanol detected in the medium by gas chromatography. The concentration of 3-methyl-1-butanol was reduced from 122.0 to 48.9 muM when 5.0 mM valine was supplemented to limiting leucine cultures. The amino acids isoleucine, threonine, norleucine, norvaline, alpha-amino-butyrate, alanine, and glycine also spared the leucine requirement of leucine auxotrophs, most likely because they resembled leucine and competed for its uptake. We propose that leucine analogs restrict the entry and degradation of leucine and thus reduce its conversion to 3-methyl-1-butanol, a major component of fusel oil. PMID- 16346214 TI - Prophage Origin of a Virulent Phage Appearing on Fermentations of Lactobacillus casei S-1. AB - For protection from the abnormal fermentation of Lactobacillus casei S-1 caused by contamination of a virulent phage, phiFSV, the origin of this phage was studied. Morphologies, viral structural proteins, and DNA structures of three independent isolates of phiFSV were compared with those of phiFSW, which is lysogenized in strain S-1. The results showed (i) that the morphology of phiFSV phages is indistinguishable from that of phiFSW and (ii) that all viral structural components found in phiFSW are present in the particles of phiFSV's. In addition, restriction endonuclease analyses of viral DNA showed that the HindIII-digested fragments of phiFSW DNA, the sum of which covered at least 94.7% of this phage genome, were conserved in the phiFSV DNA digests. Results of Southern filter hybridization of the S-1 and prophage-cured cell (C239) DNAs with phiFSV DNA as a probe revealed that C239 had lost most of the phiFSV DNA sequence, whereas S-1 had about one copy of the phiFSV DNA sequence. These results indicate that virulent phage phiFSV is derived from the lysogenized phage phiFSW. Therefore, the appearance of phiFSV can be eliminated by using the prophage-cured derivative of S-1. PMID- 16346215 TI - Isolation and Characterization of a Methylotrophic Marine Methanogen, Methanococcoides methylutens gen. nov., sp. nov. AB - A new genus of marine methanogenic bacteria is described that utilizes trimethylamine, diethylamine, monomethylamine, and methanol as substrates for growth and methanogenesis. Methane was not produced from H(2)-CO(2), sodium formate, or sodium acetate. Growth on trimethylamine was stimulated by yeast extract, Trypticase (BBL Microbiology Systems, Cockeysville, Md.), rumen fluid, or B vitamins. The optimal growth temperature was 30 to 35 degrees C. The maximum growth rate was between pH 7.0 and 7.5. Na (0.4 M) and MgSO(4) (0.05 M) were required for maximum growth. Colonies of the type strain, TMA-10, were yellow, circular, and convex with entire edges. Cells were nonmotile, nonsporeforming, irregular cocci 1 mum in diameter which stained gram negative and occurred singly or in pairs. Micrographs of thin sections revealed a monolayered cell wall approximately 10-nm thick which consisted of protein. Cells were lysed in 0.01% sodium dodecyl sulfate or 0.001% Triton X-100. The DNA base composition was 42 mol% guanine plus cytosine. Methanococcoides is the proposed genus and Methanococcoides methylutens is the type species. TMA-10 is the type strain (ATCC 33938). PMID- 16346216 TI - Methanogenic bacteria from the bondyuzhskoe oil field: general characterization and analysis of stable-carbon isotopic fractionation. AB - Selective enrichment culture techniques were employed to obtain mixed cultures of methanogenic rods and sarcina from surface flooding waters and deep subsurface ( approximately 1650 m) oil-bearing sedimentary rocks and formation waters sampled from an old oil field in the U.S.S.R. previously reported to display active biological methanogenesis. The methanogens were selectively isolated as colonies on agar petri dishes that were incubated in a novel container. The general cellular and growth features of three Methanobacterium isolates were determined. These strains grew optimally at 37 to 45 degrees C in anaerobic pressure tube cultures with a doubling time of 16 to 18 h on H(2)-CO(2) and proliferated as autotrophs. Acetate addition significantly enhanced the final cell yield. Growth of these strains was completely inhibited by either 0.6 g of sodium sulfide per liter or 31.0 of sodium chloride per liter, but growth was not inhibited by either 0.3 g of sodium sulfide per liter or 1.0 g of sodium sulfate per liter. One novel isolate, Methanobacterium sp. strain ivanov, was grown on H(2)-CO(2), and the stable-carbon isotopic fractionations that occurred during synthesis of methane, cell carbon, and lipids were determined. The results of this study were used to examine the anomalous relationship between the isotopic and chemical compositions of natural gas occurring in the deep subsurface environment of the oil field. PMID- 16346217 TI - Method for rapid detection of cyanogenic bacteria. AB - An agar plate method is described in which the production of hydrogen cyanide by as many as 50 microbial isolates per plate may be detected. Cyanide produced by the organisms reacts with copper(II) ethylacetoacetate and 4,4'-methylenebis-(N,N dimethylaniline) in a paper disk suspended above the microbial colonies. Cell growth occurs in depressions in the agar surface, which allows separation of colonies and enhances sensitivity of hydrogen cyanide detection. PMID- 16346218 TI - Survival of Rhizobium phaseoli in Coal-Based Legume Inoculants Applied to Seeds. AB - Eight coals used as carriers in legume inoculants promoted the survival of Rhizobium phaseoli on pinto bean seeds. Although peat was more protective, most coal-based inoculants provided >10 viable rhizobia per seed after 4 weeks. PMID- 16346219 TI - Replica plating method for estimating phenanthrene-utilizing and phenanthrene cometabolizing microorganisms. AB - A replica plating method was developed for detecting and enumerating phenanthrene degrading microorganisms. The method is designed to discriminate between aquatic organisms that utilize phenanthrene as the sole carbon and energy source and organisms that cometabolize phenanthrene. The method was used to demonstrate that phenanthrene utilizers and phenanthrene cometabolizers coexist in estuarine sediments. PMID- 16346220 TI - Free-living Rhizobium strain able to grow on n(2) as the sole nitrogen source. AB - A Rhizobium strain isolated from stem nodules of the legume Sesbania rostrata was shown to grow on atmospheric nitrogen (N(2)) as the sole nitrogen source. Non N(2)-fixing mutants isolated directly on agar plates formed nodules that did not fix N(2) when inoculated into the host plant. PMID- 16346221 TI - Adaptation of Rhodopseudomonas sphaeroides to Growth on d-(-)-Tartrate and Large Scale Production of a Constitutive d-(-)-Tartrate Dehydratase During Growth on dl Malate. AB - Of 10 strains of the purple non-sulfur bacterium Rhodopseudomonas sphaeroides, 8 acquired the ability to grow on d-(-)-tartrate; however, growth occurred only after extended lag phases ranging from 2 to 14 days. These lag phases occurred because only a small number of inoculum cells were able to grow by forming the enzyme d-(-)-tartrate dehydratase [d-(-)-tartrate hydro-lyase; EC number not yet available]. Once cells had grown on d-(-)-tartrate, d-(-)-tartrate dehydratase was formed constitutively. Therefore, mass cultivation of R. sphaeroides for production of large quantities of enzyme was possible on substrates much cheaper than d-(-)-tartrate. When 0.38 mol of dl-malate was used as a substrate in a chemotrophic fed-batch culture, a final biomass of 15 g (dry weight) liter and 1,500 U of d-(-)-tartrate dehydratase liter of culture were formed. The enzyme can be used for selective cleavage of racemic tartaric acid and for quantitative determination of d-(-)-tartrate. PMID- 16346222 TI - Effect of Sugar Concentration in Jerusalem Artichoke Extract on Kluyveromyces marxianus Growth and Ethanol Production. AB - The effect of inulin sugars concentration on the growth and ethanol production by Kluyveromyces marxianus UCD (FST) 55-82 was studied. A maximum ethanol concentration of 102 g/liter was obtained from 250 g of sugars per liter initial concentration. The maximum specific growth rate varied from 0.44 h at 50 g of sugar per liter to 0.13 h at 300 g of sugar per liter, whereas the ethanol yield remained almost constant at 0.45 g of ethanol per g of sugars utilized. PMID- 16346224 TI - Detection of industrial pollutants and toxic chemical wastes in sewage treatment plant influents by use of a biological monitor. AB - Heterogeneous mixtures of toxic chemical wastes affecting the normal operations of a sewage treatment facility employing anaerobic digestion were detected by dark-field observation of uncoordinate flagellar orientation patterns displayed by cells of Spirillum volutans. These results were compared with plant parameters determined by gas production, volatile fatty acids, and alkalinity measurements. PMID- 16346223 TI - Nutritional interdependence among rumen bacteria during cellulose digestion in vitro. AB - A study has been made of the promoting effect of starch on cellulose digestion by mixed rumen bacteria in a cellulose-urea medium. Starch supplementation of the medium promoted the growth of bacteria that required neither amino acids (AA) nor branched-chain fatty acids (BrFA). The growth of these bacteria was followed by the growth of AA-dependent bacteria, AA- or BrFA-dependent bacteria, BrFA producing bacteria, and finally, BrFA-dependent cellulolytic bacteria. Population changes of these bacterial groups corresponded with a cross-feeding of AA and BrFA and the overall disappearance of cellulose. The data suggest that the nutritional interdependence among rumen bacteria affects the rate of cellulose digestion. PMID- 16346225 TI - Effect of Antecedent Growth Conditions on Sensitivity of Escherichia coli to Chlorine Dioxide. AB - [This corrects the article on p. 818 in vol. 44.]. PMID- 16346226 TI - Effects of Iron Starvation on the Ultrastructure of the Cyanobacterium Agmenellum quadruplicatum. AB - The effects of iron starvation on the ultrastructure of the unicellular cyanobacterium Agmenellum quadruplicatum were studied by using thin sectioning and transmission electron microscopy. Intracellular polysaccharide began to accumulate at the onset of iron limitation. This was followed by degradation of ribosomes and (later) degradation of the thylakoid membranes, both of which were virtually absent by 200 h. The thylakoids underwent structural modifications and rearrangements before they actually began to break down. Iron starvation did not appear to affect carboxysomes or the extracellular glyocalyx. On the other hand, polyphosphate bodies may have been partially degraded, and an electrontransparent gap eventually appeared between the cell wall and the cytoplasmic membrane. All of these changes were reversed when iron was added back to 200-h starved cultures. The sequence of ultrastructural changes observed during iron starvation clearly differed from those previously reported to occur during nitrogen, phosphorous, or carbon limitation. PMID- 16346227 TI - Construction and Characterization of Isogenic Series of Saccharomyces cerevisiae Polyploid Strains. AB - Tetraploid cells of Saccharomyces cerevisiae are generated spontaneously in a homothallic MATa/MATalpha diploid population at low frequency (approximately 10 per cell) through the homozygosity of mating-type alleles by mitotic recombination followed by homothallic switching of the mating-type alleles. To isolate tetraploid clones more effectively, a selection method was developed that used a dye plate containing 40 mg each of eosin Y and amaranth in synthetic nutrient agar per liter. It was possible to isolate tetraploid clones on the dye plate at a frequency of 1 to 3% among the colonies colored dark red in contrast to the light red of the original diploid colonies. Isogenic series of haploid to tetraploid clones with homozygous or heterozygous genomic configurations were easily constructed with the tetraploid strains. No significant differences in specific growth rate or fermentative rate were observed corresponding to differences in ploidy, although the haploid clones showed a higher frequency of spontaneous respiratory-deficient cells than did the others. However, a significant increment in the fermentative rate in glucose nutrient medium was observed in the hybrid strains constructed with two independent homozygous cell lines. These observations strongly suggest that the polyploid strains favored by the brewing and baking industries perform well not because of the physical increment of the cellular volume by polyploidy but because of the genetic complexity or heterosis by heterozygosity of the genome in the hybrid polyploid cells. PMID- 16346228 TI - Starvation-survival processes of a marine Vibrio. AB - Levels of DNA, RNA, protein, ATP, glutathione, and radioactivity associated with [S]methionine-labeled cellular protein were estimated at various times during the starvation-survival process of a marine psychrophilic heterotrophic Vibrio sp., Ant-300. Values for the macromolecules were analyzed in terms of total, viable, and respiring cells. Electron micrographs (thin sections) were made on log-phase and 5.5-week-starved cells. On a per-cell basis, the levels of protein and DNA rapidly decreased until a constant level was attained. A second method in which radioactive sulfur was used for monitoring protein demonstrated that the cellular protein level decreased for approximately 2.5 weeks and then remained constant. An initial decrease in the RNA level with starvation was noted, but with time the RNA (orcinol-positive material) level increased to 2.5 times the minimum level. After 6 weeks of starvation, 45 to 60% of the cells remained capable of respiration, as determined by iodonitrotetrazolium violet-formazan granule production. Potential respiration and endogenous respiration levels fell, with an intervening 1-week peak, until at 2 weeks no endogenous respiration could be measured; respiratory potential remained high. The cell glutathione level fell during starvation, but when the cells were starved in the presence of the appropriate amino acids, glutathione was resynthesized to its original level, beginning after 1 week of starvation. The cells used much of their stored products and became ultramicrocells during the 6-week starvation-survival process. Ant-300 underwent many physiological changes in the first week of starvation that relate to the utilization or production of ATP. After that period, a stable pattern for long-term starvation was demonstrated. PMID- 16346229 TI - In Vitro Survey of Autographa californica Nuclear Polyhedrosis Virus Interaction with Nontarget Vertebrate Host Cells. AB - Thirty-five nontarget host cell lines, 23 of human and 12 of nonhuman vertebrate origin, were exposed to Autographa californica nuclear polyhedrosis virus preparations derived from four different sources: polyhedra, hemolymph, cell culture medium, and cultured cells. The virus and cells were incubated together at two different temperatures, 28 or 37 degrees C, for four different lengths of time, 16, 40, 64, or 168 h, and the cells were assayed for the presence of virus by a peroxidase-antiperoxidase detection method. The estimated sensitivity of the assay as routinely conducted was 0.98 ng of alkali-liberated viral protein and 1.95 ng of budded viral protein per mm. No evidence of frank replication was obtained in any of the 35 cell lines tested, although virus uptake appeared to be quite common. Virus uptake was confirmed in some cases by electron microscopy. The degree of virus uptake appeared to be dependent on cell type, time and temperature of incubation, and viral phenotype. Virus purified from polyhedra was generally taken up more readily than were the other forms tested. PMID- 16346230 TI - Diagenesis of metals chemically complexed to bacteria: laboratory formation of metal phosphates, sulfides, and organic condensates in artificial sediments. AB - Cells of Bacillus subtilis, when suspended in a 5mM metal solution, bind metals tenaciously to their cell walls. These metal-loaded cells, when mixed with a synthetic sediment and put under laboratory conditions to simulate low temperature sediment diagenesis, nucleate the formation of a mixed assemblage of crystalline metal phosphates, metal sulfides, and polymeric, metal-complexed, organic residues. The sequential series of diagenetic events leading to the formation of authigenic mineral phases was followed by transmission electron microscopy and energy-dispersive X-ray analysis. The minerals quartz (SiO(2)) and calcite (CaCO(3)) were employed in the synthetic sediment. Crystalline magnetite (Fe(2)O(3)) and elemental sulfur were added as redox buffering agents to ensure anoxic conditions. Quartz and magnetite appeared unreactive throughout the experimental conditions. Elemental sulfur interacted with the metal-loaded cells, affected both the eventual chemistry and crystal habit of the metal phosphates, and formed a variety of crystalline metal sulfides. Calcite raised the pH of the fluid phase of the sediment, which influenced phosphate mineralization and inhibited metal sulfide genesis. PMID- 16346231 TI - Starvation-survival patterns of sixteen freshly isolated open-ocean bacteria. AB - Sixteen marine isolates from a NORPAX cruise, which were transferred once on medium after initial isolation, survived nutrient deprivation for at least 8 months (longest period test). All but one isolate remained cellularly intact, although their sizes and shapes changed greatly, and all became smaller, decreasing in size from 40 to 79%. Three starvation-survival patterns were demonstrated, namely (i) an initial increase in viable cells followed by a decrease until a constant number was reached, (ii) an increase in viable cells until a constant number was reached, and (iii) a decrease in viable cells until a constant number was reached. One isolate from each starvation-survival pattern was starved for 8 months and then was tested in comparison with 4-month-starved Ant-300 for [C]glutamic acid uptake, respiration, and incorporation. The response to glutamic acid was rapid and linear in each case. The data indicate that the starvation-survival of Ant-300 is not an anomalous situation and that open ocean bacteria can withstand nutrient deprivation for long periods of time and still retain the capacity for active metabolism, if the nutrients become available. PMID- 16346232 TI - Microbial treatment of soil to remove pentachlorophenol. AB - Direct inoculation of bacteria capable of degrading pentachlorophenol (PCP) into PCP-contaminated soil was investigated as a prophylactic measure to reduce the hazards of runoffs when spills occur or when wooden poles freshly treated with PCP-containing preservatives are located near streams and lakes. In laboratory tests at 30 degrees C, the direct addition of 10 PCP-utilizing Arthrobacter cells per g of dry soil reduced the half-life of the pesticide from 2 weeks to <1 day. Soil inoculation also was shown to be an effective way to increase the PCP disappearance rate in a test conducted in an outdoor shed. PMID- 16346233 TI - Effects of selected assay parameters on measurement of lignocellulose mineralization with a radiolabeled substrate. AB - Varying the amount of labeled substrate or the amount of leaf material resulted in significant nonlinear changes in lignocellulose mineralization, as measured with natural [C]lignin-labeled lignocellulose. The use of periodic rather than continuous aeration was found not to have significant effects on measured mineralization. PMID- 16346234 TI - Rapid method for detection and quantitation of hydroxylated aromatic intermediates produced by microorganisms. AB - Tetrazotized o-dianisidine was used for quantitative measurements of the amount of 1-naphthol formed from naphthalene by fungi in liquid cultures. The reagent was also used to detect the accumulation of phenols and cis-dihydrodiols in fungal and bacterial colonies on solid media. PMID- 16346235 TI - Production of Epoxides from alpha,beta-Halohydrins by Flavobacterium sp. AB - The relative activity of Flavobacterium whole cells on the enzymatic synthesis of epoxides from alpha,beta-chlorohydrins, -bromohydrins, and -iodohydrins is described. PMID- 16346236 TI - Effect of phenol molecular structure on bacterial transformation rate constants in pond and river samples. AB - Microbial transformation rate constants for a series of phenols were correlated with a property of the substituents, van der Waal's radius. Transformation products were the corresponding catechols, with the exception of p-hydroxybenzoic acid, the product of p-acetylphenol. A different product suggested a different pathway; p-acetylphenol, therefore, was deleted from the data base. PMID- 16346237 TI - Acetone, Isopropanol, and Butanol Production by Clostridium beijerinckii (syn. Clostridium butylicum) and Clostridium aurantibutyricum. AB - Thirty-four strains representing 15 species of anaerobic bacteria were screened for acetone, isopropanol, and n-butanol (solvent) production. Under our culture conditions, several strains of Clostridium beijerinckii and C. aurantibutyricum produced at least 40 mM n-butanol (C. acetobutylicum strains produced up to 41 mM n-butanol under similar conditions). Both solvent-producing and non-solvent producing strains of C. beijerinckii have high DNA homology with a reference strain of C. beijerinckii. Strains labeled "Clostridium butylicum" are phenotypically similar to C. beijerinckii and showed at least 78% DNA homology to a reference strain of C. beijerinckii. Therefore, these "C. butylicum" strains are members of C. beijerinckii. An earlier DNA homology study has shown that C. beijerinckii, C. aurantibutyricum, and C. acetobutylicum are distinct species. PMID- 16346238 TI - Metabolism of ochratoxin a by rats. AB - [This corrects the article on p. 785 in vol. 44.]. PMID- 16346239 TI - Identification of Rhizobium phaseoli Strains That Are Tolerant or Sensitive to Soil Acidity. AB - A study was conducted to determine whether the survival of Rhizobium phaseoli in acid soils could be predicted on the basis of the tolerance of the organism to acidity in culture. Of 16 strains tested, all grew in culture at pH 4.6, but only those that grew at pH 3.8 survived in soils having pH values of 4.1 to 4.6. Strains that tolerated the lowest pH values in culture were tolerant of the highest aluminum concentrations. In one acid soil, an acid-tolerant strain was unable to survive in numbers greater than 100/g, but the poor survival was not related to the level of extractable aluminum or manganese in the soil. Reproduction of an acid-tolerant strain of R. phaseoli was enhanced in the rhizosphere of Phaseolus vulgaris in both acid and limed soils, but stimulation of an acid-sensitive strain by the plant occurred only in the limed soil. These results indicate that cultural tests can be used to predict the ability of R. phaseoli to survive in acid soil. PMID- 16346240 TI - Reduced sulfur in ashes and slags from the gasification of coals: availability for chemical and microbial oxidation. AB - This study was initiated to determine if reduced sulfur contained in coal gasifier ash and slag was available for microbial and chemical oxidation because eventual large-quantity landfill disposal of these solid wastes is expected. Continuous application of distilled water to a column containing a high-sulfur content (4% [wt/wt]) gasifier slag yielded leachates with high sulfate levels (1,300 mg of sulfate liter) and low pH values (4.2). At the end of the experiment, a three-tube most-probable-number analysis indicated that the waste contained 1.3 x 10 thiosulfate-oxidizing bacteria per g. Slag samples obtained aseptically from the column produced sulfate under both abiotic and biotic conditions when incubated in a mineral nutrient solution. Both microbial and chemical sulfate syntheses were greatly stimulated by the addition of thiosulfate to the slag-mineral nutrient solution. These results led to a test of microbial versus chemical sulfur oxidation in ashes and slags from five gasification processes. Sulfate production was measured in sterile (autoclaved) and nonsterile suspensions of the solid wastes in a mineral nutrient solution. These ashes and slags varied in sulfur content from 0.3 to 4.0% (wt/wt). Four of these wastes demonstrated both chemical (2.0 to 27 mug of sulfate g day) and microbial (3.1 to 114 mug of sulfate g day) sulfur oxidation. Obvious relationships between sulfur oxidation rate and either sulfur content or particle size distribution of the wastes were not immediately evident. We conclude that the sulfur contained in all but one waste is available for oxidation to sulfuric acid and that microorganisms play a partial role in this process. PMID- 16346241 TI - Oxygen Microelectrode That Is Insensitive to Medium Chemical Composition: Use in an Acid Microbial Mat Dominated by Cyanidium caldarium. AB - A novel oxygen microelectrode with a tip diameter of 2 to 20 mum was constructed which could function satisfactorily under a variety of environmental conditions and in a variety of media, including human blood serum, citric acid at pH 2.5, moist air, and paraffin oil. Measurement of oxygen by this electrode does not require stirring of the medium. Electrodes could be made so that the 90% response time necessary to detect changes in oxygen concentration was less than 0.2 s, and response was linear with oxygen concentration. To demonstrate the performance of the electrode, oxygen and photosynthesis profiles of an acid microbial mat (pH 2.8) dominated by the eucaryotic alga Cyanidium caldarium were made. Photosynthetic rates as high as 95 mmol of O(2) dm h were measured within the most active 0.1-mm layer, which was ca. 0.2 mm below the surface of the microbial mat. The total photosynthetic activity was 47 mmol of O(2) m h. Vertical profiles of photosynthesis at different light intensities showed that the microalgae within the mat were not photoinhibited at bright sunlight (2,090 muEinsteins m s). PMID- 16346242 TI - Bacterial methylation of chlorinated phenols and guaiacols: formation of veratroles from guaiacols and high-molecular-weight chlorinated lignin. AB - Two strains of bacteria, provisionally assigned to the genus Arthrobacter, were shown to metabolize mono-, di-, tri-, and tetrachloroguaiacols and pentachlorophenol to the corresponding O-methyl compounds. Hydroxylated intermediates were formed only transiently, except for the synthesis by one strain of 3,4,5-trichlorosyringol from 3,4,5-trichloroguaiacol. Two isomeric trichloroveratroles and tetrachloroveratrole were formed by three of the strains from a high-molecular-weight chlorinated lignin isolated from kraft pulp mill bleach plant. The concentrations of methylated metabolites varied widely and did not appear to be correlated with degradation. The possible environmental consequences resulting from synthesis of these highly lipophilic substances are discussed briefly. PMID- 16346243 TI - Influence of substratum wettability on attachment of freshwater bacteria to solid surfaces. AB - We studied the attachment of a number of freshwater bacteria from River Sowe, Coventry, England, to test substrata. The attachment of each organism to hydrophobic and hydrophilic surfaces was evaluated, and further studies evaluated the attachment of selected isolates to a number of substrata with a range of water wettabilities. The wettability of each substratum was determined by contact angle measurements and was expressed as the work of adhesion (W(A)). No generic pattern of attachment to the test surfaces was found, although the majority of the organisms isolated showed a preference for the hydrophobic surface. A more detailed study of selected isolates showed a relationship between W(A) and number of attached cells. Each bacterium attached in maximum numbers to a surface that was characteristic of that organism and that had a W(A) between 75 and 105 mJ m. PMID- 16346244 TI - Amino Acid assimilation and electron transport system activity in attached and free-living marine bacteria. AB - Amino acid assimilation and electron transport system activity of a marine Pseudomonas sp. was evaluated to determine whether the activity of bacteria attached to solid surfaces differed from that of free-living bacteria or bacteria which had been attached but subsequently desorbed from the substratum (detached bacteria). Bacteria were allowed to attach to glass and to a range of plastic surfaces (Thermanox, polyvinylidene fluoride, polyethylene, polytetrafluoroethylene). Microautoradiography and staining with a tetrazolium salt to demonstrate electron transport system activity were used to compare the activity of these organisms with that of free-living or detached cells. The water wettability of the surfaces was evaluated by measuring the advancing contact angle (theta(A)) of water on each surface, to determine whether there was a relationship between activity and substratum hydrophilicity. There was an increase in the proportion of leucine-assimilating attached bacteria and in the proportion of attached cells demonstrating electron transport system activity with an increase in substratum theta(A), but the relationship between activity of attached and free-living cells depended on the substratum. Activity appeared to promote firm attachment, and detached bacteria assimilated fewer amino acids than did attached cells. There was no general effect of surfaces on attached bacterial activity, and attached cells may be more, or less, active than free-living cells, depending on the amino acid, its concentration, and substratum properties. PMID- 16346245 TI - Mineralization of parathion in the rice rhizosphere. AB - We studied CO(2) evolution from ring-labeled [2,6-C]parathion (O,O-diethyl-O-p nitrophenyl phosphorothioate) in the rhizosphere of rice seedlings. The soil samples (nonflooded [60% water-holding capacity] and flooded) were treated first with technical parathion (20 mug/g) and then after 10 days with ring-labeled [C]parathion. In unplanted soil, less than 5.5% of the C in the parathion was evolved as CO(2) in 15 days under both flooded and nonflooded conditions. In soil planted with rice, 9.2% of the radiocarbon was evolved as CO(2) under nonflooded conditions, and 22.6% was evolved under flooded conditions. These results suggest that soil planted with rice permits significant ring cleavage, especially under flooded conditions. PMID- 16346246 TI - Effects of Nitrogen Supplements on Degradation of Aspen Wood Lignin and Carbohydrate Components by Phanerochaete chrysosporium. AB - A supplement of KH(2)PO(4), MgSO(4), CaCl(2), trace elements, and thiamine accelerated the initial rate of aspen wood decay by Phanerochaete chrysosporium but did not increase the extent of lignin degradation. Asparagine, casein hydrolysate, and urea supplements (1% added N) strongly inhibited lignin degradation and weight loss. The complex nitrogen sources peptone and yeast extract stimulated lignin degradation and weight loss. Albumen and NH(4)Cl had intermediate effects. Conversion of [C]lignin to CO(2) and water-soluble materials underestimated lignin degradation in the presence of the complex N sources. The highest ratio of lignin degradation to total weight loss and the largest increase in cellulase digestibility occurred during the decay of unsupplemented wood. Rotting of aspen wood by P. chrysosporium gives smaller digestibility increases than have been found with some other white-rot fungi. PMID- 16346247 TI - Effects of Nitrogen Sources on Cellulose and Synthetic Lignin Degradation by Phanerochaete chrysosporium. AB - Phanerochaete chrysosporium degraded cellulose faster with organic nitrogen sources than with NH(4)Cl. Simple and complex nitrogen sources added at the time of inoculation to N-limited cultures of P. chrysosporium, with glucose as carbon/energy source, transiently stimulated degradation of synthetic [C]lignin to CO(2). The same nitrogen sources added 5 days after inoculation, when the cultures were entering secondary metabolism, delayed CO(2) production. The various N sources affected synthetic lignin degradation in defined medium differently than lignin degradation in aspen wood. PMID- 16346248 TI - Hydrogen Recycling by Rhizobium leguminosarum Isolates and Growth and Nitrogen Contents of Pea Plants (Pisum sativum L.). AB - The ability to recycle H(2) evolved by nitrogenase is thought to be of importance in increasing the efficiency of N(2) fixation and to be a factor in increasing plant yield in symbiotic systems. To determine whether this ability is a significant factor in the Rhizobium leguminosarum-Pisum sativum L. system, plants were inoculated with R. leguminosarum isolates which differed in their ability to oxidize H(2) and in their relative efficiency of N(2) fixation. These plants were grown at three levels of irradiance and harvested after 3, 4, and 5 weeks of growth for determination of C(2)H(2) reduction, H(2) evolution and uptake, plant dry weight, and N content. Plants inoculated with uptake hydrogenase-positive (Hup) isolates did not exhibit higher dry weight or N content than those inoculated with Hup isolates under any of the growth conditions studied. The efficiency of the nitrogenase system of Hup isolates increased at a low irradiance, a factor which may allow them to compete successfully with Hup isolates. In some HupR. leguminosarum isolates, H(2) oxidation is coupled to ATP formation, whereas in others, it is not. There were no differences in plant dry weight and N content in plants inoculated with the two types and grown for 5 weeks at three irradiance levels. The addition of H(2) to Hup nodules whose supply of photosynthate had been removed by stem excision did not increase C(2)H(2) reduction in either coupled or uncoupled types. PMID- 16346249 TI - Effects of Fusariotoxin T-2 on Saccharomyces cerevisiae and Saccharomyces carlsbergensis. AB - A Fusarium metabolite, T-2 toxin, inhibits the growth of Saccharomyces carlsbergensis and Saccharomyces cerevisiae. The growth inhibitory concentrations of T-2 toxin were 40 and 100 mug/ml, respectively, for exponentially growing cultures of the two yeasts. S. carlsbergensis was more sensitive to the toxin and exhibited a biphasic dose-response curve. Addition of the toxin at 10 mug/ml of S. carlsbergensis culture resulted in a retardation of growth as measured turbidimetrically, after only 30 to 40 min. This action was reversible upon washing the cells free of the toxin. The sensitivity of the yeasts to the toxin was dependent upon the types and concentrations of carbohydrates used in the growth media. The sensitivity of the cells to the toxin decreased in glucose repressed cultures. These results suggest that T-2 toxin interferes with mitochondrial functions of these yeasts. PMID- 16346250 TI - Light as an energy source in continuous cultures of bacteriorhodopsin-containing halobacteria. AB - The role of light as an energy source for slightly aereated cultures of halobacteria was studied, using continuous cultures with low nutrient concentrations and a low oxygen supply. A series of experiments were carried out with non-illuminated and differently illuminated cultures and with different oxygen transfer rates. Under low oxygen availability, light proved to be a decisively important energy source that allowed the populations to reach higher growth rates and much higher population densities. Oxygen influenced the growth over only a minimal level, below which neither the illuminated nor the dark cultures were affected by the oxygen transfer rate. From these results, it appears that the bacteriorhodopsin-mediated energy supply could have a very important role for the ecology of halobacteria in their microaerophilic habitats. In the illuminated cultures, cells that originated purple colonies on plates appeared. These cells, which could be bacteriorhodopsin-constitutive mutants, are now being studied. PMID- 16346251 TI - Microbial Resolution of alpha-Hydroxy Acids by Enantiospecifically Dehydrogenating Bacteria from Soil. AB - Thirty-three bacterial strains were isolated from soil, utilizing optically asymmetric degradation of dl-2-hydroxy-4-methylpentanoic acid (dl-HMPA) as the screening probe. Those strains were distributed in the following group and genera: Coryneform and Bacillus, Pseudomonas, and Streptomyces. Among them, the most potent strains, Bacillus freudenreichii NRS-137KH20B and Brevibacterium albidum NRS-130KH20B, could perform the resolution of more than 30 g of dl-HMPA per liter within 4 to 5 days of fermentation. Optically pure l- and d-HMPA enantiomers were obtained in more than 80% theoretical yield, whereas the transformed enantiomer was almost quantitatively recovered as 2-oxo-4-methyl pentanoic acid in the culture broth. The enantiospecific dehydrogenation responsible for this resolution reaction had a rather wide substrate specificity on straight or branched aliphatic C(4) to C(16) 2-hydroxy acids, exhibiting the optima at chain lengths of either C(7) or C(5), although the enantiospecificity was not changed by chain length. The process was thus successfully extended to the preparation of optically pure C(5) to C(9) 2-hydroxy acids. PMID- 16346252 TI - Rapid Colony Screening Method for Identifying Hydrogenase Activity in Rhizobium japonicum. AB - A method has been developed for the rapid screening of Rhizobium japonicum colonies for hydrogenase activity based on their ability to reduce methylene blue in the presence of respiratory inhibitors and hydrogen. Hydrogen uptake-positive (Hup) colonies derepressed for hydrogenase activity were visualized by their localized decolorization of filter paper disks impregnated with the dye. Appropriate responses were seen with a number of Hup and Hup wild-type strains of R. japonicum as well as Hup mutants. Its specificity was further confirmed in selected strains on the basis of comparisons with chemolithotrophic growth and the presence of other genetic markers. Utilization of the method in identifying Hup colonies among 16,000 merodiploid derivatives of the Hup mutant strain PJ17nal containing cloned DNA fragments of the Hup strain 122 DES has demonstrated its applicability as a screening procedure in the genetic analysis of the R. japonicum hydrogen uptake system. PMID- 16346253 TI - Lignin Degradation by Streptomyces viridosporus: Isolation and Characterization of a New Polymeric Lignin Degradation Intermediate. AB - A new, quantitatively significant intermediate formed during lignin degradation by Streptomyces viridosporus T7A was isolated and characterized. In Streptomyces inoculated cultures, the intermediate, an acid-precipitable, polyphenolic, polymeric lignin (APPL), accumulated in the growth medium. The APPL was a water soluble polymer probably consisting of a heterogeneous mixture of molecular weight components of >/=20,000. APPLs were precipitable from culture filtrates after they had been acidified to pH <3 to 5. Noninoculated controls yielded little APPL, but supernatant solutions from inoculated cultures produced quantities of APPL that correlated with the biodegradability of the lignocellulose type. Maximal recovery of APPL was obtained from corn lignocellulose, reaching 30% of the initial lignin present in the substrate. APPLs contained small amounts of carbohydrate, organic nitrogen, and inorganic materials. The lignin origin of APPLs was confirmed by chemical analyses, which included acidolysis, permanganate oxidation, elemental analyses, functional group analyses, nuclear magnetic resonance spectroscopy, and C isotopic techniques. Analyses of APPLs from corn lignocelluloses showed that S. viridosporus-degraded APPLs were lignin derived but significantly different in structure from APPLs derived from uninoculated controls or from a standard corn milled-wood lignin. Degraded APPLs were enriched in phenolic hydroxyl groups and, to a small extent, in carboxyl groups. Degradative changes appeared to be largely oxidative and were thought to involve substantial cleavage of p-hydroxy ether linkages and methoxyl groups in the lignin. PMID- 16346254 TI - Production and Characteristics of Raw Starch-Digesting Glucoamylase O from a Protease-Negative, Glycosidase-Negative Aspergillus awamori var. kawachi Mutant. AB - Production of a raw starch-digesting glucoamylase O (GA O) by protease-negative, glycosidase-negative mutant strain HF-15 of Aspergillus awamori var. kawachi was undertaken under submerged culture conditions. The purified GA O was electrophoretically homogeneous and similar to the parent glucoamylase I (GA I) in the hydrolysis curves toward gelatinized potato starch, raw starch, and glycogen and in its thermostability and pH stability, but it was different in molecular weight and carbohydrate content (250,000 and 24.3% for GA O, 90,000 and ca. 7% for GA I, respectively). The chitin-bound GA O hydrolyzed raw starch but the chitin-bound GA I failed to digest raw starch because chitin was adsorbed at the raw starch affinity site of the GA I molecule. The removal of the raw starch affinity site of GA O with subtilisin led to the formation of a modified GA O (molecular weight, 170,000), which hydrolyzed glycogen 100%, similar to GA O and GA I, and was adsorbed onto chitin and fungal cell wall but not onto raw starch, Avicel, or chitosan. The modified GA I (molecular weight, 83,000) derived by treatment with substilisin hydrolyzed glycogen up to only 80% and failed to be adsorbed onto any of the above polysaccharides. The N-bromosuccinimide-oxidized GA O lost its activity toward gelatinized and raw starches, but the abilities to be adsorbed onto raw starch and chitin were preserved. It was thus suggested that both the raw starch affinity site essential for raw starch digestion and the chitin-binding site specific for the binding with chitin in the cell wall could be different from the active site, located in the three respective positions in the GA O molecule. PMID- 16346255 TI - Chlorobium limicola forma thiosulfatophilum: Biocatalyst in the Production of Sulfur and Organic Carbon from a Gas Stream Containing H(2)S and CO(2). AB - Chlorobium limicola forma thiosulfatophilum (ATCC 17092) was grown in a 1-liter continuously stirred tank reactor (800-ml liquid volume) at pH 6.8, 30 degrees C, saturated light intensity, and a gas flow rate of 23.6 ml/min from a gas cylinder blend consisting of 3.9 mol% H(2)S, 9.2 mol% CO(2), 86.4 mol% N(2), and 0.5 mol% H(2). This is the first demonstration of photoautotrophic growth of a Chlorobium sp. on a continuous inorganic gas feed. A significant potential exists for applying this photoautotrophic process to desulfurization and CO(2) fixation of gases containing acidic components (H(2)S and CO(2)). PMID- 16346256 TI - Production and Characterization of Amylase from Calvatia gigantea. AB - alpha-Amylase (EC 3.2.1.1) was excreted by Calvatia gigantea in liquid growth media containing different sources of starch. Among the factors affecting enzyme production in shake flasks were the type and the concentration of starch and the nitrogen source supplied. Optimum cultural conditions for maximum enzyme production were: soluble starch concentration, 5%; inoculum size, 3.75 x 10 conidia per ml; 5-day cultivation time at 28 to 30 degrees C. The observed maximum yield of 81.3 U of saccharifying enzyme activity per ml of growth medium was the highest ever reported in the literature for submerged cultures. Partially purified enzyme functioned optimally at pH 4.5 to 5.5 and 53 to 58 degrees C. The activation energy of enzymic hydrolysis of starch in the range of 20 to 40 degrees C was 8,125 cal/mol (ca. 3.41 x 10 J). The apparent K(m) value of the enzyme at 25 degrees C was 7.68 x 10 g/ml. Some of the properties of the enzyme under investigation were similar to those of alpha-amylases excreted from molds producing large amounts of the enzyme. PMID- 16346257 TI - Competition Among Rhizobium leguminosarum Strains for Nodulation of Lentils (Lens esculenta). AB - Thirty-one cultures of Rhizobium leguminosarum were screened for effectiveness (C(2)H(2) reduction) on lentils (Lens esculenta). Fluorescent antibodies prepared against three of the most effective strains (Hawaii 5-0, Nitragin 92A3, and Nitragin 128A12) exhibited a high degree of strain specificity; the antibodies reacted strongly with their homologous rhizobia in culture and with bacteroids in nodules. They did not cross-react with one another, and only weakly with 5 of the 47 other R. leguminosarum cultures tested. In competition studies in the growth chamber, whenever strain Nitragin 92A3 was included in the inoculum mixture, it consistently (but not always significantly, P = 0.05) occupied the majority of nodules on all four cultivars used. However, some degree of strain X cultivar interaction was apparent: Hawaii 5-0 was of equal competitiveness (P = 0.05) with Nitragin 92A3 on three of the varieties (Commercial, Tekoa, and Benewah), but inferior (P = 0.01) on the Chilean variety; Nitragin 92A3 completely dominated (P = 0.01) Nitragin 128A12 on all cultivars; and Hawaii 5-0 was of equal competitiveness (P = 0.05) to Nitragin 128A12 on the Chilean variety and more competitive (P = 0.01) on the commercial variety and less so on the other two varieties. In field experiments, Hawaii 5-0 proved of equal competitiveness (P = 0.01) with Nitragin 92A3 in one soil (an Inceptisol) and superior (P 2NH(3) + H(2) which gives a conversion factor between C(2)H(2) and N(2) of 4. It is proposed that, in general, 4 is a more appropriate value than 3 for the conversion factor. PMID- 16346273 TI - Method for measuring changes in surface tension on agar. AB - The surface tension of agar surfaces was determined by measuring the contact angles formed by drops of various hydrophobic liquids on the surface and then calculating the composite surface free energy function by solving a series of simultaneous equations derived from these data. This method was used to measure the change in the surface tension of agar produced by the addition of various concentrations of albumin. The resulting curve was typical of the effect of increasing concentrations of surfactants on surface tension. The method was compared with other methods of determining surface tension of solids, and it was concluded that the technique used here provided the most reliable results. PMID- 16346274 TI - Regulation of expression of nitrate and dinitrogen assimilation by anabaena species. AB - Anabaena sp. strain 7120 appeared more responsive to nitrogen control than A. cylindrica. Growth in the presence of nitrate strongly repressed the differentiation of heterocysts and fixation of dinitrogen in Anabaena sp. strain 7120, but only weakly in A. cylindrica. Nitrate assimilation by ammonium-grown cultures was strongly repressed in Anabaena sp. strain 7120, but less so in A. cylindrica. The repressive effect of nitrate on dinitrogen assimilation in Anabaena sp. strain 7120, compared to A. cylindrica, did not correlate with a greater rate of nitrate transport, reduction to ammonium, assimilation into amino acids, or growth. Although both species grew at similar rates with dinitrogen, A. cylindrica grew faster with nitrate, incorporated more NO(3) into amino acids, and assimilated (transported) nitrate at the same rate as Anabaena sp. strain 7120. Full expression of nitrate assimilation in the two species occurred within 2.5 h (10 to 14% of their generation times) after transfer to nitrate medium. The induction and continued expression of nitrate assimilation was dependent on protein synthesis. The half-saturation constants for nitrate assimilation and for nitrate and ammonium repression of dinitrogen assimilation have ecological significance with respect to nitrogen-dependent growth and competitiveness of the two Anabaena species. PMID- 16346275 TI - Ensifer adhaerens Predatory Activity Against Other Bacteria in Soil, as Monitored by Indirect Phage Analysis. AB - An indirect phage analysis procedure was used to detect and follow the activity of the bacterial predator Ensifer adhaerens in situ in natural soil. The soil was percolated with an aqueous suspension of washed bacterial host cells so that the E. adhaerens cells naturally present in the soil would multiply in response to the host cells. The natural phage development which ensued against these multiplying E. adhaerens cells in the soil was then monitored by noting plaques which developed when the percolation fluid was plated with laboratory strains of E. adhaerens on laboratory media. The activities of the other members of the predation system that includes E. adhaerens (Streptomyces sp. strain 34 and a myxobacter) could not be monitored directly by phage analysis because phage were not found for them. Indirect monitoring was possible, however, because they were susceptible to attack by E. adhaerens. In general, the results were in agreement with previous observations by other methods of the predation sequence. E. adhaerens attacked Micrococcus luteus, Streptomyces sp. strain 34, and the myxobacter but did not attack several other possible species of hosts. It also did not respond to percolation of the soil with various nutrient solutions. E. adhaerens phage activity was not present in half of the soils percolated with M. luteus cells. This seemed to reflect too great a phage-host specificity for the technique as regards these soils, because E. adhaerens-like bacteria other than the strains used for plaquing were present in at least some of these soils. Although E. adhaerens did not attack Escherichia coli or Pseudomonas aeruginosa in soil, there was an overproduction of E. adhaerens phage if these bacteria were percolated simultaneously with M. luteus cells. The possibility is discussed that this represents an activation by M. luteus (or by a heat-extractable factor from it) of other bacterial predators that attack E. coli or P. aeruginosa and that these predators subsequently are themselves attacked by E. adhaerens. PMID- 16346276 TI - Sporulation of Clostridium acetobutylicum P262 in a Defined Medium. AB - A defined minimal sporulation medium for Clostridium acetobutylicum P262, which produces high levels of solvents, is described. The overall sporulation sequence was similar to that of other endospore-forming bacteria. However, we observed a presporulation stage, during which swollen phase-bright cells which contained large amounts of granulose formed. During sporulation, the initiation of spore coat formation occurred before the onset of cortex formation. Other Clostridium strains tested showed marked variations in ability to grow and sporulate in various minimal media. PMID- 16346277 TI - Occurrence of the Rumen Ciliate Oligoisotricha bubali in Domestic Cattle (Bos taurus). AB - Oligoisotricha bubali, previously observed twice in water buffalo, was detected in rumen contents of domestic cattle (Bos taurus) in two different areas of Tennessee. Concentrations ranged from <1 to 35% of the total protozoa in unweaned calves and up to 72% in older animals in feedlot. In contrast to the other genera of holotrichs, both total numbers and percent composition of O. bubali increased when animals were fed a corn silage-concentrate diet. PMID- 16346278 TI - Aggregate stabilization of volcanic ash and soil during microbial degradation of straw. AB - Bacterial and fungal colonists of straw in soil promoted aggregate stabilization of volcanic ash and soil. PMID- 16346279 TI - Regiochemistry of Camphor Analog Oxidation by Pseudomonas putida. AB - Pseudomonas putida cooxidized norcamphor and pericyclocamphanone to hydroxylated and lactonized products during growth on camphor. Norcamphor was hydroxylated at the 5 position, similar to the corresponding process in camphor, but pericyclocamphanone was oxidized at the 6 position. We conclude that the regiochemistry of the hydroxylation may be substrate controlled. PMID- 16346280 TI - Effects of 2-bromoethanesulfonic Acid and 2- chloroethanesulfonic Acid on acetate utilization in a continuous-flow methanogenic fixed-film column. AB - 2-Bromoethanesulfonic acid (BESA) and 2-chloroethanesulfonic acid (CESA) have been reported to be potent inhibitors of methane formation during methanogenic decomposition in batch cultures. However, in a laboratory-scale continuous-flow methanogenic fixed-film column containing a predominance of acetate decarboxylating methanogens, BESA at 6 x 10 M produced only a 41% inhibition of acetate utilization, and CESA at 5.4 x 10 M produced a 37% inhibition of acetate utilization. BESA and CESA concentrations were not monitored in the effluent, so their fate is unknown. The organisms in the column were capable of degrading trace halogenated aliphatic compounds ( approximately 30 mug/liter) with acetate (100 mg/liter) as the primary substrate. Previous exposure of the cells to halogenated organic compounds may have conferred resistance to BESA and CESA. Degradation of the inhibitor compounds is another possible explanation for the observed effects. PMID- 16346281 TI - Pathways of propionate degradation by enriched methanogenic cultures. AB - A mixed methanogenic culture was highly enriched in a growth medium containing propionate as the sole organic carbon and energy source. With this culture, the pathways of propionate degradation were studied by use of C-radiotracers. Propionate was first metabolized to acetate, carbon dioxide, and hydrogen by nonmethanogenic organisms. Formate was not excreted. The carbon dioxide originated exclusively from the carboxyl group of propionate, whereas both [2-C]- and [3-C]propionate lead to the production of radioactive acetate. The methyl and carboxyl groups of the acetate produced were equally labeled, regardless of whether [2-C]- or [3-C]propionate was used. These observations suggest that in the culture, propionate was degraded through a randomizing pathway. PMID- 16346282 TI - Phytol degradation by marine bacteria. AB - Microbial degradation of phytol is often proposed to be the primary source of the acyclic isoprenoid acids observed in sediments, yet only a limited number of these acids have been found in bacterial cultures grown on phytol. This study reports detailed capillary gas chromatography and gas chromatography-mass spectrometry analyses of the products resulting from growth of marine bacteria on phytol as the sole carbon source. We examined two strains of bacteria which were able to oxidize phytol to phytenic acid but were unable to further degrade phytol. The third isolate studied converted phytol to a mixture of five saturated isoprenoid acids. The C(17) isoprenoid acid produced was of particular interest, since its genesis from phytol would have involved several unusual intermediates. It is suggested that this acid is produced by bacterial metabolism of the C(18) isoprenoid ketone (produced from phytol abiologically under oxic conditions) and that its abundance is thus a sensitive indicator of sedimentary depositional conditions. PMID- 16346283 TI - Selection of wine yeasts for growth and fermentation in the presence of ethanol and sucrose. AB - To optimize the conversion of carbohydrates to ethanol, strains of several Saccharomyces species were examined for the ability to grow and ferment in a range of sucrose and ethanol concentrations. A total of 632 wine yeasts, most of them isolated from wineries in Andalusia and Extremadura, southwestern Spain, were subjected to screening and selection. Growth and fermentative capacity in different ethanol and sucrose concentrations varied from one strain to another. There was no correlation between growth and fermentative capacity. The best 35 strains grew in 15% ethanol and fermented in 18% ethanol. Ethanol accumulated, although at a reduced rate, after the cells stopped growing. Most yeast strains were highly fermentative in 50% sucrose. Some of them effectively utilized the carbohydrates of the culture, yielding final ethanol concentrations of > 14%. Of the 35 selected strains, 16 were promising for genetic analysis and breeding because of their capacity to sporulate. These strains were homothallic, and their spores were viable. The meiotic products analyzed so far were also homothallic. PMID- 16346284 TI - Reductive dehalogenations of halobenzoates by anaerobic lake sediment microorganisms. AB - Methane-producing freshwater lake sediment was found to dehalogenate chloro-, bromo-, and iodobenzoates by a reductive reaction in which the halogen was replaced by a hydrogen atom. The identity of the dehalogenated products was confirmed by mass spectrometry, nuclear magnetic resonance, or cochromatography. Removal of the halogens to produce benzoate was necessary before mineralization to CH(4) + CO(2) could occur. The dehalogenation occurred after a lag period which lasted from 1 week to more than 6 months, depending on the chemical. Dehalogenation was not observed in the absence of CH(4) production, and it was inhibited by the addition of 20% O(2). Once sediment was acclimated to halobenzoate dehalogenation, new additions of the halobenzoate were degraded without lag. Acclimation was observed regardless of whether the parent substrates were eventually mineralized to CH(4) + CO(2). Sediment acclimated to bromo- and chlorobenzoate degradation generally metabolized bromo- and chlorobenzoates, but sediment acclimated to iodobenzoate degradation only metabolized iodobenzoate. Prior acclimation of sediment to benzoate decomposition did not alter the pattern of dehalogenation, and sediment acclimated to dehalogenation was not concurrently acclimated to benzoate degradation. The presence of this apparent specificity, the lag period, and subsequent acclimation, together with our findings of the absence of dehalogenation in sterile sediments and by sediments previously incubated at >/=39 degrees C, suggests that this reaction was biologically catalyzed. Apparently, a pathway for the reductive dehalogenation of aryl halides is present in anaerobic microorganisms of this methanogenic sediment. PMID- 16346285 TI - Kinetics of microbial dehalogenation of haloaromatic substrates in methanogenic environments. AB - The kinetic parameters associated with the microbial dehalogenation of 3 chlorobenzoate, 3,5-dichlorobenzoate, and 4-amino-3,5-dichlorobenzoate were measured in anoxic sediment slurries and in an enriched methanogenic culture grown on 3-chlorobenzoate. The initial dehalogenation of the substrates exhibited Michaelis-Menten kinetics. The apparent K(m) values for the above substrates ranged from 30 to 67 muM. The pattern of degradation, however, was unusual. The enrichment culture accumulated partially dehalogenated intermediates to 72 and 98% of that possible when incubated with either 3,5-dichloro- or 4-amino-3,5 dichlorobenzoate, respectively, but did not accumulate significant amounts of benzoate when 3-chlorobenzoate was the sole carbon and energy source. The accumulated intermediates were rapidly metabolized only after the parent substrate concentrations were nearly depleted (<5 muM). A sequential Michaelis Menten model was developed to account for the observed pattern of biodegradation. Using this model, we found that relative differences in the K(m) and V(max) parameters for substrate and intermediate dehalogenations alone were insufficient to explain the transitory accumulation of intermediates. However, by inserting a competitive inhibition term, with the primary substrate as the inhibitor, the observed pattern of degradation was simulated. Apparently, the dichlorinated substrates competitively inhibit the dehalogenation of the monochlorinated substrates. Similar kinetic patterns were noted for sediments, although the rates were slower than in the enrichment culture. PMID- 16346286 TI - Entrainment of viruses from septic tank leach fields through a shallow, sandy soil aquifer. AB - A study was conducted which focused on movement of naturally occurring human enteroviruses from a subsurface wastewater disposal system through a shallow aquifer. The potential for significant entrainment of virus particles was evidenced by their recovery at down-gradient distances of 67.05 m and from aquifer depths of 18 m. A significant negative correlation was observed between virus occurrence and the distance from the "septage" (leaching pool) source. Virus occurrence could not be statistically correlated with either total or fecal coliforms, indicating the limitations of current microbial water quality indicators for predicting the virological quality of groundwater. PMID- 16346287 TI - Frequencies of Bacteriophage-Resistant and Slow Acid-Producing Variants of Streptococcus cremoris. AB - The frequencies of bacteriophage-resistant and slow acid-producing variants in 10 purified strains of Streptococcus cremoris were studied. There were considerable differences among the strains in the occurrence of both slow acid-producing and phage-resistant mutants. Nevertheless, the spontaneous rates of mutation to slow acid production were three to five orders of magnitude greater than the corresponding rates of mutation to phage resistance, suggesting that slow acid production and phage resistance are not genetically linked, although they appear in cultures concomitantly. The frequencies of slow acid-producing variants among resistant and sensitive isolates from the same parent culture were similar and appeared to be strain dependent. All phage-resistant mutants tested were found to be deficient in adsorption of the homologous bacteriophage. PMID- 16346288 TI - Radioassay for hydrogenase activity in viable cells and documentation of aerobic hydrogen-consuming bacteria living in extreme environments. AB - An isotopic tracer assay based on the hydrogenase-dependent formation of tritiated water from tritium gas was developed for in life analysis of microbial hydrogen transformation. This method allowed detection of bacterial hydrogen metabolism in pure cultures or in natural samples obtained from aquatic ecosystems. A differentiation between chemical-biological and aerobic-anaerobic hydrogen metabolism was established by variation of the experimental incubation temperature or by addition of selective inhibitors. Hydrogenase activity was shown to be proportional to the consumption or production of hydrogen by cultures of Desulfovibrio vulgaris, Clostridium pasteurianum, and Methanosarcina barkeri. This method was applied, in connection with measurements of free hydrogen and most-probable-number enumerations, in aerobic natural source waters to establish the activity and document the ecology of hydrogen-consuming bacteria in extreme acid, thermal, or saline environments. The utility of the assay is based in part on the ability to quantify bacterial hydrogen transformation at natural hydrogen partial pressures, without the use of artificial electron acceptors. PMID- 16346289 TI - Hydrogen metabolism by decomposing cyanobacterial aggregates in big soda lake, nevada. AB - Hydrogen production by incubated cyanobacterial epiphytes occurred only in the dark, was stimulated by C(2)H(2), and was inhibited by O(2). Addition of NO(3) inhibited dark, anaerobic H(2) production, whereas the addition of NH(4) inhibited N(2) fixation (C(2)H(2) reduction) but not dark H(2) production. Aerobically incubated cyanobacterial aggregates consumed H(2), but light incubated rates (3.6 mumol of H(2) g h) were statistically equivalent to dark uptake rates (4.8 mumol of H(2) g h), which were statistically equivalent to dark, anaerobic production rates (2.5 to 10 mumol of H(2) g h). Production rates of H(2) were fourfold higher for aggregates in a more advanced stage of decomposition. Enrichment cultures of H(2)-producing fermentative bacteria were recovered from freshly harvested, H(2)-producing cyanobacterial aggregates. Hydrogen production in these cyanobacterial communities appears to be caused by the resident bacterial flora and not by the cyanobacteria. In situ areal estimates of dark H(2) production by submerged epiphytes (6.8 mumol of H(2) m h) were much lower than rates of light-driven N(2) fixation by the epiphytic cyanobacteria (310 mumol of C(2)H(4) m h). PMID- 16346290 TI - Broad-Host-Range Agrocin of Agrobacterium tumefaciens. AB - Eighteen strains of Agrobacterium tumefaciens isolated from crown galls were tested for agrocin production. Of six agrocin-producing strains, one (D286) produced a broad-host-range agrocin active against strains carrying nopaline, octopine, and agropine type Ti plasmids. Sensitivity to agrocin D286 was found to map in the 11- to 18-megadalton region of the nopaline Ti plasmid pTiC58. The agrocin was partially purified, and its physical characteristics were consistent with its being a nucleotide, as is agrocin 84. Agrocin D286 was shown to inhibit DNA, RNA, and protein syntheses. Strain D286 spontaneously lost its pathogenicity, and its potential for use in the biological control of crown gall is discussed. PMID- 16346291 TI - Microbiological Survey of Adirondack Lakes with Various pH Values. AB - Nine high-altitude oligotrophic Adirondack lakes in upstate New York having water of pH 4.3 to 7.0 were surveyed for total bacterial numbers and possible adaptation of the microbial communities to environmental pH. The number of heterotrophic bacteria from water samples recoverable on standard plate count agar were low (10 to 10 per ml) for most of the lakes. Acridine orange direct counts were approximately two orders of magnitude higher than plate counts for each lake. Sediment aerobic heterotrophs recovered on standard plate count agar ranged from 1.4 x 10 to 1.3 x 10 per g of sediment. Direct epifluorescence counts of bacteria in sediment samples ranged from 3.0 x 10 to 1.4 x 10 per g. Low density values were consistent with the oligotrophic nature of all the lakes surveyed. There were no apparent differences in numbers of bacteria originally isolated at pH 5.0 and pH 7.0 between circumneutral lakes (pH > 6.0) and acidic lakes (pH < 5.0). Approximately 1,200 isolates were recultured over a range of pH from 3.0 to 7.0. Regardless of the original isolation pH (pH 5.0 or pH 7.0), less than 10% of the isolates grew at pH < 5.0. Those originally isolated at pH 5.0 also grew at pH 6.0 and 7.0. Those originally isolated at pH 7.0 preferred pH 7.0, with 98% able to grow at pH 6.0 and 44% able to grow at pH 5.0. A chi-square contingency test clearly showed (P < 0.005) that two distinct heterotrophic populations had been originally isolated at pH 5.0 and pH 7.0, although there is undoubtedly some overlap between the two populations. PMID- 16346292 TI - Fermentation of Peptides by Bacteroides ruminicola B(1)4. AB - The maximum growth rate of Bacteroides ruminicola B(1)4 was significantly improved when either Trypticase or acetate and C(4)-C(5) fatty acids were added to defined medium containing macrominerals, microminerals, vitamins, hemin, cysteine hydrochloride, and glucose. The organism was unable to grow with peptides as the sole energy source, but growth yields from glucose were significantly improved when Trypticase was added to batch cultures containing basal medium, acetate, and C(4)-C(5) volatile fatty acids. During periods of rapid growth, very little peptide was deaminated to ammonia, but after growth ceased there was a linear increase in ammonia. Fifteen grams of Trypticase per liter resulted in maximum ammonia production. In glucose-limited chemostats, ammonia production from peptides was inversely proportional to the dilution rate, and 87% of the variation in ammonia production could be explained by retention time in the culture vessel. Chemostats receiving Trypticase had higher theoretical maximum growth yields and lower maintenance energy expenditures than similar cultures not receiving peptide. Cells from the Trypticase cultures contained more carbohydrate, and this difference was most evident at rapid dilution rates. When corrections were made for cell composition and the amount of peptides that were fermented, it appeared that peptide carbon skeletons could be used for maintenance energy. B. ruminicola B(1)4 was unable to grow on peptides alone because it was unable to utilize peptides at a fast enough rate to meet its maintenance requirement. PMID- 16346293 TI - Novel haloperoxidase reaction: synthesis of dihalogenated products. AB - The enzymatic synthesis of vicinal, dihalogenated products from alkenes and alkynes is described. The enzymatic reaction required an alkene or alkyne, dilute hydrogen peroxide, a haloperoxidase, and molar amounts of halide ions. Vicinal dichloro, dibromo, and diiodo products could be formed. A hydroxyl group on the carbon adjacent to the carbon-carbon double or triple bond lowered the halide ion concentration needed to produce the dihalo product. This reaction offers one explanation for the origin of natural, vicinal, dihalogenated products, such as those found frequently in marine microogranisms. PMID- 16346294 TI - Effects of metals on methanogenesis, sulfate reduction, carbon dioxide evolution, and microbial biomass in anoxic salt marsh sediments. AB - The effects of several metals on microbial methane, carbon dioxide, and sulfide production and microbial ATP were examined in sediments from Spartina alterniflora communities. Anaerobically homogenized sediments were amended with 1,000 ppm (ratio of weight of metal to dry weight of sediment) of various metals. Time courses in controls were similar for CH(4), H(2)S, and CO(2), with short initial lags (0 to 4 h) followed by periods of constant gas production (1 to 2 days) and declining rates thereafter. Comparisons were made between control and experimental assays with respect to initial rates of production (after lag) and overall production. Methane evolution was inhibited both initially and overall by CH(3)HgCl, HgS, and NaAsO(2). A period of initial inhibition was followed by a period of overall stimulation with Hg, Pb, Ni, Cd, and Cu, all as chlorides, and with ZnSO(4), K(2)CrO(4), and K(2)Cr(2)O(7). Production of CO(2) was generally less affected by the addition of metals. Inhibition was noted with NaAsO(2), CH(3)HgCl, and Na(2)MoO(4). Minor stimulation of CO(2) production occurred over the long term with chlorides of Hg, Pb, and Fe. Sulfate reduction was inhibited in the short term by all metals tested and over the long term by all but FeCl(2) and NiCl(2). Microbial biomass was decreased by FeCl(2), K(2)Cr(2)O(7), ZnSO(4), CdCl(2), and CuCl(2) but remained generally unaffected by PbCl(2), HgCl(2), and NiCl(2). Although the majority of metals produced an immediate inhibition of methanogenesis, for several metals this was only a transient phenomenon followed by an overall stimulation. The initial suppression of methanogenesis may be relieved by precipitation, complexation, or transformation of the metal (possibly by methylation), with the subsequent stimulation resulting from a sustained inhibition of competing organisms (e.g., sulfate-reducing bacteria). For several environmentally significant metals, severe metal pollution may substantially alter the flow of carbon in sediments. PMID- 16346295 TI - Physiology of Ex Planta Nitrogenase Activity in Rhizobium japonicum. AB - Thirty-nine wild-type strains of Rhizobium japonicum have been studied for their ability to synthesize nitrogenase ex planta in defined liquid media under microaerobic conditions. Twenty-one produced more than trace amounts of acetylene reduction activity, but only a few of these yielded high activity. The oxygen response curves were similar for most of the nitrogenase-positive strains. The strains derepressible for activity had several phenotypic characteristics different from non-derepressible strains. These included slower growth and lower oxygen consumption under microaerobic conditions and lower extracellular polysaccharide production. Extracellular polysaccharide production during growth on gluconate in every nitrogenase-positive strain assayed was lower under both aerobic and microaerobic conditions than the non-derepressible strains. These phenotypic characteristics may be representative of a genotype of a subspecies of R. japonicum. These studies were done in part to enlarge the base number of strains available for studies on the physiology, biochemistry, and genetics of nitrogen fixation. PMID- 16346296 TI - Volatile Fatty Acid production by the hindgut microbiota of xylophagous termites. AB - Acetate dominated the extracellular pool of volatile fatty acids (VFAs) in the hindgut fluid of Reticulitermes flavipes, Zootermopsis angusticollis, and Incisitermes schwarzi, where it occurred at concentrations of 57.9 to 80.6 mM and accounted for 94 to 98 mol% of all VFAs. Small amounts of C(3) to C(5) VFAs were also observed. Acetate was also the major VFA in hindgut homogenates of Schedorhinotermes lamanianus, Prorhinotermes simplex, Coptotermes formosanus, and Nasutitermes corniger. Estimates of in situ acetogenesis by the hindgut microbiota of R. flavipes (20.2 to 43.3 nmol . termite . h) revealed that this activity could support 77 to 100% of the respiratory requirements of the termite (51.6 to 63.6 nmol of O(2) . termite . h). This conclusion was buttressed by the demonstration of acetate in R. flavipes hemolymph (at 9.0 to 11.6 mM), but not in feces, and by the ability of termite tissues to readily oxidize acetate to CO(2). About 85% of the acetate produced by the hindgut microbiota was derived from cellulose C; the remainder was derived from hemicellulose C. Selective removal of major groups of microbes from the hindgut of R. flavipes indicated that protozoa were primarily responsible for acetogenesis but that bacteria also functioned in this capacity. H(2) and CH(4) were evolved by R. flavipes (usually about 0.4 nmol . termite . h), but these compounds represented a minor fate of electrons derived from wood dissimilation within R. flavipes. A working model is proposed for symbiotic wood polysaccharide degradation in R. flavipes, and the possible roles of individual gut microbes, including CO(2)-reducing acetogenic bacteria, are discussed. PMID- 16346297 TI - Bacterioplankton in antarctic ocean waters during late austral winter: abundance, frequency of dividing cells, and estimates of production. AB - Bacterioplankton productivity in Antarctic waters of the eastern South Pacific Ocean and Drake Passage was estimated by direct counts and frequency of dividing cells (FDC). Total bacterioplankton assemblages were enumerated by epifluorescent microscopy. The experimentally determined relationship between in situ FDC and the potential instantaneous growth rate constant (mu) is best described by the regression equation ln mu = 0.081 FDC - 3.73. In the eastern South Pacific Ocean, bacterioplankton abundance (2 x 10 to 3.5 x 10 cells per ml) and FDC (11%) were highest at the Polar Front (Antarctic Convergence). North of the Subantarctic Front, abundance and FDC were between 1 x 10 to 2 x 10 cells per ml and 3 to 5%, respectively, and were vertically homogeneous to a depth of 600 m. In Drake Passage, abundance (10 x 10 cells per ml) and FDC (16%) were highest in waters south of the Polar Front and near the sea ice. Subantarctic waters in Drake Passage contained 4 x 10 cells per ml with 4 to 5% FDC. Instantaneous growth rate constants ranged between 0.029 and 0.088 h. Using estimates of potential mu and measured standing stocks, we estimated productivity to range from 0.62 mug of C per liter . day in the eastern South Pacific Ocean to 17.1 mug of C per liter . day in the Drake Passage near the sea ice. PMID- 16346298 TI - Initial effects of the mount st. Helens eruption on nitrogen cycle and related chemical processes in ryan lake. AB - Ryan Lake, a 1.6-hectare basin lake near the periphery of the tree blowdown area in the blast zone 19 km north of Mount St. Helens, was studied from August to October 1980 to determine the microbial and chemical response of the lake to the eruption. Nutrient enrichment through the addition of fresh volcanic material and the organic debris from the surrounding conifer forest stimulated intense microbial activity. Concentrations of such nutrients as phosphorus, sulfur, manganese, iron, and dissolved organic carbon were markedly elevated. Nitrogen cycle activity was especially important to the lake ecosystem in regulating biogeochemical cycling owing to the limiting abundance of nitrogen compounds. Nitrogen fixation, both aerobic and anaerobic, was active from aerobic benthic and planktonic cyanobacteria with rates up to 210 nmol of N(2) cm h and 667 nmol of N(2) liter h, respectively, and from anaerobic bacteria with rates reaching 220 nmol of N(2) liter h. Nitrification was limited to the aerobic epilimnion and littoral zones where rates were 43 and 261 nmol of NO(2) liter day, respectively. Potential denitrification rates were as high as 30 mumol of N(2)O liter day in the anaerobic hypolimnion. Total bacterial numbers ranged from 1 x 10 to 3 x 10 ml with the number of viable sulfur-metal-oxidizing bacteria reaching 2 x 10 ml in the hypolimnion. A general scenario for the microbial cycling of nitrogen, carbon, sulfur, and metals is presented for volcanically impacted lakes. The important role of nitrogen as these lakes recover from the cataclysmic eruption and proceed back towards their prior status as oligotrophic alpine lakes is emphasized. PMID- 16346299 TI - Light and electron microscopic studies of microorganisms growing in rotating biological contactor biofilms. AB - The biofilms growing in the first compartments of two rotating biological contactors used to treat municipal wastewater were examined by light and electron microscopy. The biofilms were found to contain a complex and varied microbial community that included filamentous and unicellular bacteria, protozoa, metazoa, and (possibly) bacteriophage. The predominant microorganism among these appeared to be a filamentous bacterium that was identical to Sphaerotilus in both morphological and ultrastructural characteristics. It was possible to isolate a Sphaerotilus-like bacterium from each contactor. Both the Sphaerotilus filaments and the wide variety of unicellular bacteria present tended to contain poly-beta hydroxybutyrate inclusions, a probable indication that these organisms were removing carbon from the wastewater and storing it. The microbial population of the biofilms appeared to be metabolically active, as evidenced by the presence of microcolonies and dividing cells. PMID- 16346300 TI - Isolation and Partial Characterization of Two Aeromonas hydrophila Bacteriophages. AB - Two Aeromonas hydrophila bacteriophages, Aeh1 and Aeh2, were isolated from sewage. Both phages showed binal symmetry. The dimensions of A. hydrophila phages Aeh1 and Aeh2 differed from those of the other Aeromonas phages. Also, phage Aeh2 was the largest Aeromonas phage studied to date. Phage Aeh1 formed small, clear plaques, and phage Aeh2 formed turbid plaques with clear centers. Both phages were sensitive to chloroform treatment, being totally inactivated after treatment for 1 h at 60 degrees C at pH 3 and 11. However, the infectivity of Aeh1 phage stocks increased by approximately fivefold after they were treated at pH 10 for 1 h at 22 degrees C. Phages Aeh1 and Aeh2 were serologically unrelated and had latent periods of 39 and 52 min, respectively. The average burst sizes of phages Aeh1 and Aeh2 were 17 and 92 PFU per cell, respectively. Phage Aeh1 infected 13 of 22 A. hydrophila strains tested, whereas phage Aeh2 infected only its original host. Phage Aeh1 infected some A. hydrophila strains only at or below 37 degrees C. Neither phage infected the two A. (Plesiomonas) shigelloides strains used in this study. PMID- 16346302 TI - Microbial Transformations of Natural Antitumor Agents: Products of Rotenone and Dihydrorotenone Transformation by Cunninghamella blakesleeana. AB - [This corrects the article on p. 617 in vol. 45.]. PMID- 16346301 TI - Isolation of the siderophore schizokinen from soil of rice fields. AB - Schizokinen, a siderophore previously isolated from low-iron cultures of Bacillus megaterium and Anabaena sp., was isolated from the soil of rice fields after the annual flooding period. PMID- 16346303 TI - Aerobic, Selenium-Utilizing Bacillus Isolated from Seeds of Astragalus crotalariae. AB - [This corrects the article on p. 738 in vol. 44.]. PMID- 16346305 TI - l-Serine Production by a Mutant of Sarcina albida Defective in l-Serine Degradation. AB - For improved l-serine production, an l-serine dehydratase-defective mutant of Sarcina albida IAM 1012 was obtained. In the mutant, the activities of the enzymes responsible for l-serine production were as high as those in the parent strain, and, at a low glycine concentration, the mutant accumulated l-serine more efficiently than the parent. Under optimum conditions, 21 mg of l-serine per ml accumulated from 100 mg of glycine per ml. l-Serine was isolated from a reaction mixture as l-serine m-xylene-4-sulfonate, and free amino acid was obtained in high yields by use of an ion-exchange resin. Residual glycine was recovered at a yield of 61%. PMID- 16346304 TI - Estimating Bacterioplankton Production by Measuring [H]thymidine Incorporation in a Eutrophic Swedish Lake. AB - Bacterioplankton abundance, [H]thymidine incorporation, CO(2) uptake in the dark, and fractionated primary production were measured on several occasions between June and August 1982 in eutrophic Lake Norrviken, Sweden. Bacterioplankton abundance and carbon biomass ranged from 0.5 x 10 to 2.4 x 10 cells liter and 7 to 47 mug of C liter, respectively. The average bacterial cell volume was 0.185 mum. [H]thymidine incorporation into cold-trichloroacetic acid-insoluble material ranged from 12 x 10 to 200 x 10 mol liter h. Bacterial carbon production rates were estimated to be 0.2 to 7.1 mug of C liter h. Bacterial production estimates from [H]thymidine incorporation and CO(2) uptake in the dark agreed when activity was high but diverged when activity was low and when blue-green algae (cyanobacteria) dominated the phytoplankton. Size fractionation indicated negligible uptake of [H]thymidine in the >3-mum fraction during a chrysophycean bloom in early June. We found that >50% of the H activity was in the >3-mum fraction in late August; this phenomenon was most likely due to Microcystis spp., their associated bacteria, or both. Over 60% of the CO(2) uptake in the dark was attributed to algae on each sampling occasion. Algal exudate was an important carbon source for planktonic bacteria. Bacterial production was roughly 50% of primary production. PMID- 16346306 TI - Influence of Environmental Factors and Medium Composition on Vibrio gazogenes Growth and Prodigiosin Production. AB - Vibrio gazogenes ATCC 29988 growth and prodigiosin synthesis were studied in batch culture on complex and defined media and in chemostat cultures on defined medium. In batch culture on complex medium, a maximum growth rate of 0.75 h and a maximum prodigiosin concentration of 80 ng of prodigiosin . mg of cell protein were observed. In batch culture on defined medium, maximum growth rates were lower (maximum growth rate, 0.40 h), and maximum prodigiosin concentrations were higher (1,500 ng . mg of protein). In batch culture on either complex or defined medium, growth was characterized by a period of logarithmic growth followed by a period of linear growth; on either medium, prodigiosin biosynthesis was maximum during linear growth. In batch culture on defined medium, the initial concentration of glucose optimal for growth and pigment production was 3.0%; higher levels of glucose suppressed synthesis of the pigment. V. gazogenes had an absolute requirement for Na; optimal growth occurred in the presence of 100 mM NaCl. Increases in the concentration of Na up to 600 mM resulted in further increases in the concentration of pigment in the broth. Prodigiosin was synthesized at a maximum level in the presence of inorganic phosphate concentrations suboptimal for growth. Concentrations of KH(2)PO(4) above 0.4 mM caused decreased pigment synthesis, whereas maximum cell growth occurred at 1.0 mM. Optimal growth and pigment production occurred in the presence of 8 to 16 mg of ferric ion . liter, with higher concentrations proving inhibitory to both growth and pigment production. Both growth and pigment production were found to decrease with increased concentrations of p-aminobenzoic acid. The highest specific concentration of prodigiosin (3,480 ng . mg protein) was observed in chemostat cultures at a dilution rate of 0.057 h. The specific rate of prodigiosin production at this dilution rate was approximately 80% greater than that observed in batch culture on defined medium. At dilution rates greater than 0.057 h, the concentration of cells decreased with increasing dilution rate, resulting in a profile comparable to that expected for linear growth kinetics. No explanation could be found for the linear growth profiles obtained for both batch and chemostat cultures. PMID- 16346307 TI - Decolorization of Several Polymeric Dyes by the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium. AB - The polymeric dyes Poly B-411, Poly R-481, and Poly Y-606 were examined as possible alternatives to the radiolabeled lignin previously used as a substrate in lignin biodegradation assays. Like lignin degradation, the decolorization of these dyes by the white rot basidiomycete Phanerochaete chrysosporium occurred during secondary metabolism, was suppressed in cultures grown in the presence of high levels of nitrogen, and was strongly dependent on the oxygen concentration in the cultures. A variety of inhibitors of lignin degradation, including thiourea, azide, and 4'-O-methylisoeugenol, also inhibited dye decolorization. A pleiotropic mutant of P. chrysosporium, 104-2, lacking phenol oxidase and ligninolytic activity was also not able to decolorize the polymeric dyes, whereas a phenotypic revertant strain, 424-2, regained this capacity. All of these results suggest that the ligninolytic degradation activity of the fungus was responsible for the decolorization of these dyes. PMID- 16346308 TI - Protein Patterns of Growing and Starved Cells of a Marine Vibrio sp. AB - Fingerprint protein patterns were produced by two-dimensional polyacrylamide electrophoresis on lysed cells of a Vibrio sp., Ant-300, which were prepared from growing and starved cultures. The cells were labeled with [S]methionine during growth and subsequently starved for up to 30 days. Samples were taken at selected time points representing stages in the starvation-survival process. Unlabeled starved cells were allowed to recover in the presence of [S]methionine to determine protein changes associated with the recovery from starvation. All growth conditions produced similar protein fingerprints; however, some protein spots disappeared, whereas others were seen only during starvation. PMID- 16346309 TI - Heterotrophic activity throughout a vertical profile of seawater and sediment in halifax harbor, Canada. AB - The relative heterotrophic activity of marine microorganisms was determined at two sites by the heterotrophic uptake technique throughout the water column, the sediment-water interface, and the surface layer of sediment. In the water column, uptake was greatest at the surface and steadily decreased with depth. The percentage of the substrate that was respired also decreased with depth from 69 to 56%. The activity of the sediment-water interface was several orders of magnitude greater than that of the overlying water and twice that of the sediment immediately below. Hand-collected water samples carefully taken as close as 1 cm from the sediment-water interface had the same characteristically low activity as the bottom few meters of water. Microautoradiography with H-labeled glucose, glutamic acid, or thymidine revealed a general decrease in the percentage of active cells with depth from 35 to <1%. The number of active cells in the interface and sediment averaged <10% of the total population. The data indicate that the sediment-water interface is the most active region in this system due to an increased number of active cells rather than an increased percentage of active cells or increased per-cell activity. PMID- 16346310 TI - Microbial activity at the sediment-water interface in halifax harbor, Canada. AB - The sediment-water interface in Halifax Harbor supports a microbial population of 6.95 x 10 cells per g (dry weight). As determined by the standard technique of suspending subsamples in filtered seawater, the uptake of added glutamic acid by this population is 113.5 ng g (dry weight) h. An alternate technique was developed to measure the heterotrophic activity of the interface over longer periods of time, using undisturbed cores with the sediment-water interface intact. Under these conditions, the microbes in the water column and the interface increased exponentially in number, with mean doubling times of 9.6 and 4.5 days, respectively. The uptake of glutamic acid by the microbial population of the interface was determined to be 12.7 ng g (dry weight) h, almost an order of magnitude less than the uptake determined by the previous method. This indicates that substrate diffusion and competition for substrate by the microbes in the water column are important factors when considering the heterotrophic activity of the sediment microbial population. After 48 h of incubation, uptake and respiration ceased, probably due to the exhaustion of labeled substrate. Additional substrate added after 48 h of incubation was taken up at a rate similar to that measured after the first addition. It appears that the microbial population of the interface is able to respond quickly and repeatedly to relatively large nutrient additions. After 10 days of incubation, the number of "viable" cells as determined by autoradiography was much smaller than the increase in numbers as determined by direct counts. Apparently a large part of the viable population is unaffected by nutrient addition. PMID- 16346311 TI - Enhanced Mineral Uptake by Zea mays and Sorghum bicolor Roots Inoculated with Azospirillum brasilense. AB - Inoculation of corn (Zea mays) seeds with Azospirillum brasilense strain Cd or Sp 7 significantly enhanced (30 to 50% over controls) the uptake of NO(3), K, and H(2)PO(4) into 3- to 4-day- and 2-week-old root segments. No gross changes in root morphology were observed; altered cell arrangement in the outer four or five layers of the cortex was seen in photomicrographs of cross sections of inoculated corn roots. The surface activity involved in ion uptake probably increased, as shown by the darker staining by methylene blue of the affected area. Shoot dry weight increased 20 to 30% in inoculated plants after 3 weeks, presumably by enhancement of mineral uptake. Corn and sorghum plants grown to maturity on limiting nutrients in the greenhouse showed improved growth from inoculation approaching that of plants grown on normal nutrient concentrations. Enhanced ion uptake may be a significant factor in the crop yield enhancement reported for Azospirillum inoculation. PMID- 16346312 TI - Distribution of Thermus spp. in Icelandic Hot Springs and a Thermal Gradient. AB - The growth range in nature of bacteria belonging to the genus Thermus was investigated by sampling 55 different hot springs in Iceland. The springs ranged in temperature from 32 to 99 degrees C, and in pH from 2.1 to 10.1. Viable counts of Thermus spp. ranging from 10 to 10 CFU/100 ml of spring water were found in 27 of the springs sampled. The temperature range for these bacteria was found to be 55 to 85 degrees C, and the pH range was from about 6.5 to above 10. Thermus spp. were found in springs containing up to 1 mM dissolved sulfide and having conductivity up to 2,000 muS/cm. The distribution of Thermus spp. in a hot spring thermal gradient was also investigated and found to agree well with the overall distribution in individual springs. PMID- 16346313 TI - Effects of Carrier and Temperature on Survival of Rhizobium spp. in Legume Inocula: Development of an Improved Type of Inoculant. AB - The effects of inoculant carrier, temperature, and storage period on the survival of Rhizobium strains were determined by plate count and most-probable-number analyses. Preliminary experiments showed that survival of rhizobia was affected by each of these factors and their interactions. Results of further studies indicated that six strains of rhizobia survived better at high temperatures when lyophilized and suspended in an oil carrier as compared to finely ground peat. The oil base inocula contained ca. 10 viable rhizobia per g after 56 days of incubation at 60 degrees C, whereas peat base inocula contained 8 h) and were characterized by formation of both CH(4) and CO(2). Ratios of CH(4)/CO(2) from [C]trimethylamine and methylamine in uninhibited sediments indicated that a significant fraction of these substrates were catabolized via a non-methanogenic process. Data from inhibition experiments involving sodium molybdate and 2-bromoethanesulfonic acid supported this interpretation. [C]methanol was oxidized relatively slowly compared with the other substrates and was catabolized mainly to CO(2). Results from experiments with molybdate and 2-bromoethanesulfonic acid suggested that methanol was oxidized primarily through sulfate reduction. In Lowes Cove sediments, trimethylamine accounted for 35.1 to 61.1% of total methane production. PMID- 16346318 TI - Generalized Transduction in the Phytopathogen Pseudomonas syringae. AB - Bacteriophages isolated from culture supernatants of Pseudomonas syringae pv. syringae and from sewage transferred various chromosomal genes to P. syringae PS224. Linkage between arginine and tryptophan loci was demonstrated. The number of transductants recovered per milliliter was not altered appreciably by UV irradiation of selected phage isolates. In addition, the presence of the IncP2 plasmid R38 in a P. syringae PS224 arginine auxotroph did not increase the transduction frequency as it does in Pseudomonas aeruginosa. Increasing the multiplicity of infection of transducing phage Pssy15 from 1 to 10 resulted in up to a 10-fold increase in the number of transductants recovered, although the actual transductional frequency remained about the same. Treatment of transduction mixtures with DNase did not affect transductional frequency. PMID- 16346319 TI - Isolation and Partial Characterization of Bacteriophages of the Phytopathogen Pseudomonas syringae. AB - Bacteriophages isolated from culture supernatants of Pseudomonas syringae pv. syringae and from sewage were identified. The DNA from each phage was isolated and digested with the restriction endonuclease EcoRI. Eight isolates were determined to be different, with two phage isolates from sewage having restriction patterns identical to two phages from culture supernatants. The sizes of the phage DNA ranged from 24 to49 kilobases for isolates from sewage and from 39 to 52.5 kilobases for the isolates from culture supernatants. Buoyant densities of phage particles in CsCl varied from 1.498 to 1.507 g/cm for isolates from sewage and from 1.506 to 1.516 g/cm for isolates from culture supernatants. Electron microscopy revealed four morphological types. Based on plaque-forming ability of culture supernatants, 31 out of 47 strains of P. syringae are probably lysogenic. PMID- 16346320 TI - Controlling Morphological Instability of Zymomonas mobilis Strains in Continuous Culture. AB - Growth of Zymomonas mobilis ATCC 29191 and CP4 in a continuous stirred tank fermentor resulted in the selection of stable flocculating variants. Factors responsible for enhancing the system pressures selective for the morphological variants were identified. By incorporating some modifications into the design of the fermentor, it was possible to achieve steady-state operation of the chemostat with both wild-type and flocculating strains. Biochemical and microscopic studies were performed to elucidate the mechanism of flocculation in Z. mobilis. PMID- 16346321 TI - Enumeration and Localization of N(2)-Fixing Bacteria Associated with Roots of Spartina alterniflora Loisel. AB - Numbers and possible locations of N(2)-fixing bacteria were investigated in roots of Spartina alterniflora Loisel, which support nitrogenase activity in the undisturbed native habitat. N(2)-fixing bacteria were recovered in cultures both from S. alterniflora roots and from the surrounding sediment, and they formed a greater proportion of the bacteria recovered from root homogenates than from salt marsh sediment. N(2)-fixing bacteria were recovered in high numbers from the rhizoplane of S. alterniflora after roots were treated with 1 or 5% chloramine-T for 1 h or with 1% NaOCl for 1 or 2 h. Immersing S. alterniflora roots in 5% NaOCl for 1 h was more effective in distinguishing bacteria inside the roots since this treatment nearly eliminated N(2)-fixing bacteria recoverable from the rhizoplane, although high numbers of N(2)-fixing bacteria were recovered from homogenates of roots treated with 5% NaOCl for 1 h. However, this treatment was less effective with roots of Zea mays L. (Funks G4646) and Sorghum bicolor (L.) Moench (CK-60 A), indicating that techniques to surface sterilize roots should be evaluated for different plants. Bacteria were observed by light and electron microscopy inter- and intracellularly in the cortex and in the aerenchyma of S. alterniflora roots. This study clearly shows that bacteria, including N(2) fixers, colonize the interior of roots of S. alterniflora growing in a Chesapeake Bay, Maryland, salt marsh. PMID- 16346322 TI - Galactose fermentation and classification of thermophilic lactobacilli. AB - The ability to ferment galactose is a major characteristic which can be used to differentiate Lactobacillus helveticus (galactose positive) from Lactobacillus lactis and Lactobacillus bulgaricus (galactose negative). In milk cultures, galactose-positive strains produced d- and l-lactic acid with little galactose accumulation, whereas galactose-negative strains produced d-lactic acid, and galactose accumulated to high levels. PMID- 16346323 TI - Microbiological reduction of quininone to quinidine. AB - A yeast identified as Hansenula anomala var. schneggii was found capable of reducing quininone to quinidine in 50% yields in 7 days of fermentation. PMID- 16346324 TI - Adsorption of Bacteriophage eb7 on Streptococcus cremoris EB7. AB - When M17 broth was used as growth medium before preparation of cell walls, adsorption of phage eb7 on Streptococcus cremoris EB7 at pH 4.0 was stimulated. When preparation included treatment with trypsin, absorption of phage was 65%. Without trypsin treatment, absorption was 81%. Only 10 to 20% of the adsorption was irreversible. Treatment with pepsin or commercial rennet but not pure chymosin prevented adsorption on non-trypsin-treated cell walls. d-Galactosamine treatment of phage eb7 had an inactivating effect which was enhanced by l rhamnose. PMID- 16346325 TI - Plasmid Involvement in Acyclic Isoprenoid Metabolism by Pseudomonas putida. AB - An organism identified as Pseudomonas putida was found to utilize citronellol or geraniol as the sole carbon and energy source. The ability to degrade these acyclic isoprenols was associated with pSRQ50, a 50-megadalton transmissible plasmid. PMID- 16346326 TI - Simple Assay for Accurate Determination of [S]sulfate Reduction Activity. AB - A method is described for the assay of [S]sulfate reduction in which filter paper wicks are used to trap [S]sulfide. The simplicity of the technique enables large numbers of samples to be conveniently processed. Enhanced sensitivity is achieved since all acid-volatile [S]sulfides produced during the incubation period are counted. Recovery of radioactivity from added Na(2)S is excellent (mean, 100.1%; standard deviation, 1.81; n = 9) and is unaffected by sulfide concentrations of up to 400 mug per sample. Field trial results with anoxic sediment samples are presented. PMID- 16346327 TI - Comparison of verification procedures for the membrane filter total coliform technique. AB - [This corrects the article on p. 1127 in vol. 45.]. PMID- 16346328 TI - Influence of substratum wettability on attachment of freshwater bacteria to solid surfaces. AB - [This corrects the article on p. 811 in vol. 45.]. PMID- 16346329 TI - Amino Acid assimilation and electron transport system activity in attached and free-living marine bacteria. AB - [This corrects the article on p. 818 in vol. 45.]. PMID- 16346330 TI - Exogenous Valine Reduces Conversion of Leucine to 3-Methyl-1-Butanol in Saccharomyces cerevisiae. AB - [This corrects the article on p. 662 in vol. 45.]. PMID- 16346331 TI - l-Tryptophan Production by Achromobacter liquidum. AB - Conditions for the production of tryptophanase from Achromobacter liquidum and for the conversion of l-serine and indole to l-tryptophan were studied. The enzyme could be produced in amounts as great as 0.750 U/ml (degradation) and 0.294 U/ml (synthesis) by shaking cultures at 30 degrees C in a medium containing dextrin, yeast extract, l-tryptophan, and l-glutamic acid. l-Tryptophan was produced most efficiently by shaking the cells at 37 degrees C in a reaction mixture containing 60 mg of l-serine per ml, 60 mg of indole per ml, and 0.5 mM pyridoxal phosphate. After 3 days, 96 mg of l-tryptophan per ml was formed, and l tryptophan was easily isolated to 85.4% yield by concentration of the reaction mixture. PMID- 16346332 TI - Anaerobic oxalate degradation: widespread natural occurrence in aquatic sediments. AB - Significant concentrations of oxalate (dissolved plus particulate) were present in sediments taken from a diversity of aquatic environments, ranging from 0.1 to 0.7 mmol/liter of sediment. These included pelagic and littoral sediments from two freshwater lakes (Searsville Lake, Calif., and Lake Tahoe, Calif.), a hypersaline, meromictic, alkaline lake (Big Soda Lake, Nev.), and a South San Francisco Bay mud flat and salt marsh. The oxalate concentration of several plant species which are potential detrital inputs to these aquatic sediments ranged from 0.1 to 5.0% (wt/wt). In experiments with litter bags, the oxalate content of Myriophyllum sp. samples buried in freshwater littoral sediments decreased to 7% of the original value in 175 days. This suggests that plant detritus is a potential source of the oxalate within these sediments. [C]oxalic acid was anaerobically degraded to CO(2) in all sediment types tested, with higher rates evident in littoral sediments than in the pelagic sediments of the lakes studied. The turnover time of the added [C]oxalate was less than 1 day in Searsville Lake littoral sediments. The total sediment oxalate concentration did not vary significantly between littoral and pelagic sediments and therefore did not appear to be controlling the rate of oxalate degradation. However, depth profiles of [C]oxalate mineralization and dissolved oxalate concentration were closely correlated in freshwater littoral sediments; both were greatest in the surface sediments (0 to 5 cm) and decreased with depth. The dissolved oxalate concentration (9.1 mumol/liter of sediment) was only 3% of the total extractable oxalate (277 mumol/liter of sediment) at the sediment surface. These results suggest that anaerobic oxalate degradation is a widespread phenomenon in aquatic sediments and may be limited by the dissolved oxalate concentration within these sediments. PMID- 16346333 TI - Toxicity of smoke to epiphytic ice nucleation-active bacteria. AB - Wheat straw smoke aerosols and liquid smoke condensates reduced significantly both the viability and the ice-nucleating activity of Pseudomonas syringae pv. syringae and Erwinia herbicola in vitro and on leaf surfaces in vivo. Highly significant reductions in numbers of bacterial ice nuclei on the surface of both corn and almond were observed after exposure to smoke aerosols. At -5 degrees C, frost injury to corn seedlings colonized by ice nucleation-active bacteria was reduced after exposure to smoke aerosols. Effects on -9 degrees C ice nuclei, although significant, were less than on ice nuclei active at -5 degrees C. These results suggest that smoke from wildfires or smudge pots may reduce plant frost susceptibility and sources of ice nuclei important in other natural processes under some conditions. PMID- 16346334 TI - Iron-chelating compounds produced by soil pseudomonads: correlation with fungal growth inhibition. AB - Strains of Pseudomonas putida, Pseudomonas sp., and Pseudomonas aeruginosa were examined for their ability to grow in the presence of the iron chelator, ethylenediamine-di-(o-hydroxyphenylacetic acid). In vitro fungal inhibition assays showed that the isolates varied in their ability to inhibit the growth of representative fungal plant pathogens. Fungal inhibition in vitro was superior to that of previously reported Pseudomonas sp. Studies with Fusarium oxysporum forma sp. lycopersici and a susceptible tomato cultivar demonstrated that Pseudomonas putida PPU3.1 was able to significantly reduce wilt disease. PMID- 16346335 TI - Improved colorimetric determination of cell wall chitin in wood decay fungi. AB - The Svennerholm modification of the Elson-Morgan method for glucosamine analysis was evaluated for its applicability to the rapid determination of chitin in wood decay fungi. The evaluation included extent of chromogen interference, sensitivity, color stability, and hydrolysis conditions for maximum release of glucosamine from fungal cell walls. With our further modification, the Svennerholm method was shown to be suitable for rapid quantitative determination of fungal chitin without chromatographic separation of hydrolysate chromogens. PMID- 16346336 TI - Predicting 2,4-dichlorophenoxyacetic Acid ester transformation rates in periphyton-dominated ecosystems. AB - Using batch cultures, we determined transformation rate coefficients for microbial transformation of 2,4-dichlorophenoxyacetic acid butoxyethyl ester (2,4 DBE) in periphyton-dominated ecosystems. Rates of 2,4-DBE loss were measured over short periods of time (usually less than 10 h), and first-order transformation rate coefficients (k(1)) were determined under the specific conditions of low 2,4 DBE concentrations and no growth. Values for k(1) were divided by total plate counts and by biomass measured as ash-free dry weight to give second-order rate coefficients (k(b) and k(AFDW), respectively) for use in predictive models. Using periphyton attached to Teflon strips, we also determined second-order rate coefficients based on the ratio of colonized surface area to container volume (k(A)). Mean second-order rate coefficients were used to predict 2,4-DBE transformation rates in microcosms having diverse chemical and biological environments. The observed transformation rates among the microcosms were most accurately predicted by using k(A). PMID- 16346337 TI - Comparison of beta-Glucosidase Activities in Different Streptomyces Strains. AB - Cellobiase (beta-glucosidase) production was compared for two streptomycetes: Streptomyces flavogriseus, a known producer of cellulase complex, and Streptomyces sp. strain CB-12, a strain isolated for its rapid growth on cellobiose. The optimal conditions for enzyme activity were established in relation to pH, temperature, enzyme stability, and substrate affinity. The production of beta-glucosidase by the two strains depended on the carbon substrate in the medium. Cellobiose was found to repress the biosynthesis of the enzyme in S. flavogriseus and to stimulate its production in strain CB-12. The biosynthesis of the enzyme correlated well with the accumulation of glucose in the culture filtrates. The combined action of the beta-glucosidases produced by the two Streptomyces strains might allow a better utilization of the reaction products which arise during the biodegradation of cellulose. PMID- 16346338 TI - Epoxidation of short-chain alkenes by resting-cell suspensions of propane-grown bacteria. AB - Sixteen new cultures of propane-utilizing bacteria were isolated from lake water from Warinanco Park, Linden, N.J. and from lake and soil samples from Bayway Refinery, Linden, N.J. In addition, 19 known cultures obtained from culture collections were also found to be able to grow on propane as the sole carbon and energy source. In addition to their ability to oxidize n-alkanes, resting-cell suspensions of both new cultures and known cultures grown on propane oxidize short-chain alkenes to their corresponding 1,2-epoxides. Among the substrate alkenes, propylene was oxidized at the highest rate. In contrast to the case with methylotrophic bacteria, the product epoxides are further metabolized. Propane and other gaseous n-alkanes inhibit the epoxidation of propylene. The optimum conditions for in vivo epoxidation are described. Results from inhibition studies indicate that a propane monooxygenase system catalyzes both the epoxidation and hydroxylation reactions. Experiments with cell-free extracts show that both hydroxylation and epoxidation activities are located in the soluble fraction obtained after 80,000 x g centrifugation. PMID- 16346339 TI - Production of Methyl Ketones from Secondary Alcohols by Cell Suspensions of C(2) to C(4)n-Alkane-Grown Bacteria. AB - Nineteen new C(2) to C(4)n-alkane-grown cultures were isolated from lake water from Warinanco Park, Linden, N.J., and from lake and soil samples from Bayway Refinery, Linden, N.J. Fifteen known liquid alkane-utilizing cultures were also found to be able to grow on C(2) to C(4)n-alkanes. Cell suspensions of these C(2) to C(4)n-alkane-grown bacteria oxidized 2-alcohols (2-propanol, 2-butanol, 2 pentanol, and 2-hexanol) to their corresponding methyl ketones. The product methyl ketones accumulated extracellularly. Cells grown on 1-propanol or 2 propanol oxidized both primary and secondary alcohols. In addition, the activity for production of methyl ketones from secondary alcohols was found in cells grown on either alkanes, alcohols, or alkylamines, indicating that the enzyme(s) responsible for this reaction is constitutive. The optimum conditions for in vivo methyl ketone formation from secondary alcohols were compared among selected strains: Brevibacterium sp. strain CRL56, Nocardia paraffinica ATCC 21198, and Pseudomonas fluorescens NRRL B-1244. The rates for the oxidation of secondary alcohols were linear for the first 3 h of incubation. Among secondary alcohols, 2 propanol and 2-butanol were oxidized at the highest rate. A pH around 8.0 to 9.0 was found to be the optimum for acetone or 2-butanone formation from 2-alcohols. The temperature optimum for the production of acetone or 2-butanone from 2 propanol or 2-butanol was rather high at 60 degrees C, indicating that the enzyme involved in the reaction is relatively thermally stable. Metal-chelating agents inhibit the production of methyl ketones, suggesting the involvement of a metal(s) in the oxidation of secondary alcohols. Secondary alcohol dehydrogenase activity was found in the cell-free soluble fraction; this activity requires a cofactor, specifically NAD. Propane monooxygenase activity was also found in the cell-free soluble fraction. It is a nonspecific enzyme catalyzing both terminal and subterminal oxidation of n-alkanes. PMID- 16346340 TI - Degradation of Phenolic Compounds and Ring Cleavage of Catechol by Phanerochaete chrysosporium. AB - POL-88, a mutant of the white-rot fungus Phanerochaete chrysosporium, was selected for diminished phenol-oxidizing enzyme activity. A wide variety of phenolic compounds were degraded by ligninolytic cultures of this mutant. With several o-diphenolic substrates, degradation intermediates were produced that had UV spectra consistent with muconic acids. Extensive spectrophotometric and polarographic assays failed to detect classical ring-cleaving dioxygenases in cell homogenates or in extracts from ligninolytic cultures. Even so, a sensitive carrier-trapping assay showed that intact cultures degraded [U-C]catechol to [C]muconic acid, establishing the presence of a system capable of 1,2-intradiol fission. Significant accumulation of [C]muconic acid into carrier occurred only when evolution of CO(2) from [C]catechol was inhibited by treating cultures with excess nutrient nitrogen (e.g., l-glutamic acid) or with cycloheximide. l Glutamic acid is known from past work to repress the ligninolytic system in P. chrysosporium and to mimic the effect of cycloheximide. The results here indicate, therefore, that the enzyme system responsible for degrading ring cleavage products to CO(2) turns over faster than does the system responsible for ring cleavage. PMID- 16346341 TI - Microcosm and experimental pond evaluation of microbial community response to synthetic oil contamination in freshwater sediments. AB - A multivariate approach was used to evaluate the significance of synthetic oil induced perturbations in the functional activity of sediment microbial communities. Total viable cell densities, ATP-biomass, alkaline phosphatase and dehydrogenase activity, and mineralization rates of glucose, protein, oleic acid, starch, naphthalene, and phenanthrene were monitored on a periodic basis in microcosms and experimental ponds for 11 months, both before and after exposure to synthetic oil. All variables contributed to significant discrimination between sediment microbial responses in control communities and communities exposed to a gradient of synthetic oil contamination. At high synthetic oil concentrations (4,000 ml/12 m), a transient reduction in sediment ATP concentrations and increased rates of oleic acid mineralization were demonstrated within 1 week of exposure. These transient effects were followed within 1 month by a significant increase in rates of naphthalene and phenanthrene mineralization. After initial construction, both control and synthetic oil-exposed microbial communities demonstrated wide variability in community activity. All experimental microbial communities approached equilibrium and demonstrated good replication. However, synthetic oil perturbation was demonstrated by wide transient variability in community activity. This variability was primarily the result of the stimulation of polyaromatic hydrocarbon mineralization rates. In general, microcosms and pond communities demonstrated sufficient resiliency to recover from the effects of synthetic oil exposure within 3 months, although polyaromatic hydrocarbon mineralization rates remained significantly elevated. PMID- 16346342 TI - Growth and plating efficiency of methanococci on agar media. AB - Plating techniques for cultivation of methanogenic bacteria have been optimized for two members of the genus Methanococcus. Methanococcus maripaludis and Methanococcus voltae were cultivated on aerobically and anaerobically prepared agar media. Maintenance of O(2) levels below 5 mul/liter within an anaerobic glove box was necessary during plating and incubation for 90% recovery of plated cells. Under an atmosphere of H(2), CO(2), and H(2)S (79:20:1), 2 to 3 days of incubation at 37 degrees C were sufficient for the formation of visible colonies. The viability of plated cells was significantly affected by the growth phase of the culture, H(2)S concentration, and the volume of medium per plate. In addition, colony size of methanococci was affected by agar type, as well as by the concentrations of agar, H(2)S, and bicarbonate. PMID- 16346343 TI - Comparison of degradative ability, enzymatic activity, and palatability of aquatic hyphomycetes grown on leaf litter. AB - Stream fungi have the capacity to degrade leaf litter and, through their activities, to transform it into a more palatable food source for invertebrate detritivores. The objectives of the present study were to characterize various aspects of fungal modification of the leaf substrate and to examine the effects these changes have on leaf palatability to detritivores. Fungal species were grown on aspen leaves for two incubation times. Leaves were analyzed to determine the weight loss, the degree of softening of the leaf matrix, and the concentrations of ATP and nitrogen associated with leaves. The activities of a protease and 10 polysaccharide-degrading enzymes produced by each fungus were also determined. Most fungi caused similar changes in physicochemical characteristics of the leaves. All fungi exhibited the capability to depolymerize pectin, xylan, and cellulose. Differences among fungi were found in their capabilities to produce protease and certain glycosidases. Leaf palatability was assessed by offering leaves of all treatments to larvae of two caddisfly shredders (Trichoptera). Feeding preferences exhibited by the shredders were similar and indicated that they perceived distinct differences among fungi. Two fungal species were highly consumed, some moderately and others only slightly. No relationships were found between any of the fungal characteristics measured and detritivore feeding preferences. Apparently, interspecific differences among fungi other than parameters associated with biomass or degradation of structural polysaccharides influence fungal palatability to caddisfly detritivores. PMID- 16346344 TI - Partitioning of CO(2) Fixation in the Colonial Cyanobacterium Microcystis aeruginosa: Mechanism Promoting Formation of Surface Scums. AB - Constraints on inorganic carbon (C(i)) availability stimulated buoyancy in natural, photosynthetically active populations of the colonial blue-green alga (cyanobacterium) Microcystis aeruginosa. In nonmixed eutrophic river water and cultures, O(2) evolution determinations indicated C(i) limitation of photosynthesis, which was overcome either by CO(2) additions to the aqueous phase or by exposure of buoyant colonies to atmospheric CO(2). Microautoradiographs of M. aeruginosa colonies revealed partitioning of CO(2) fixation and photosynthate accumulation between peripheral and internal cells, particularly in large colonies. When illuminated colonies were suspended in the aqueous phase, peripheral cells accounted for at least 90% of the CO(2) assimilation, whereas internal cells remained unlabeled. However, when CO(2) was allowed to diffuse into colonies 15 min before illumination, a more uniform distribution of labeling was observed. Resultant differences in labeling patterns were most likely due to peripheral cells more exclusively utilizing CO(2) when ambient C(i) concentrations were low. Among colonies located at the air-water interface, internal cells showed an increased share of photosynthate production when atmospheric CO(2) was supplied. This indicated that C(i) transport was restricted in large colonies below the water surface, forcing internal cells to maintain a high degree of buoyancy, thus promoting the formation of surface scums. At the surface, C(i) restrictions were alleviated. Accordingly, scum formation appears to have an ecological function, allowing cyanobacteria access to atmospheric CO(2) when the C(i) concentration is growth limiting in the water column. PMID- 16346345 TI - Formation, Fusion, and Regeneration of Protoplasts from Wild-Type and Auxotrophic Strains of the White Rot Basidiomycete Phanerochaete chrysosporium. AB - A preparation of two commercial enzymes was used to liberate protoplasts from 16 h-old mycelium of Phanerochaete chrysosporium. Regeneration frequencies of up to 5% were attained when the protoplasts were plated in a medium containing 10% sorbose and 3% agar. Fusion of protoplasts from different auxotrophic strains in polyethylene glycol-Ca produced heterokaryons. Separation of the heterokaryons into their constituent homokaryotic strains could be effected through protoplast release and formation of colonies on regeneration agar. PMID- 16346346 TI - Ethanol Production by Saccharomyces cerevisiae Immobilized in Hollow-Fiber Membrane Bioreactors. AB - Saccharomyces cerevisiae ATCC 4126 was grown within the macroporous matrix of asymmetric-walled polysulfone hollow-fiber membranes and on the exterior surfaces of isotropic-walled polypropylene hollow-fiber membranes. Nutrients were supplied and products were removed by single-pass perfusion of the fiber lumens. Growth of yeast cells within the macrovoids of the asymmetric-walled membranes attained densities of greater than 10 cells per ml and in some regions accounted for nearly 100% of the available macrovoid volume, forming a tissue-like mass. A radial distribution of cell packing existed across the fiber wall, indicating an inadequate glucose supply to cells located beyond 100 mum from the lumen surface. By comparison, yeast cell growth on the exterior surfaces of the isotropic-walled membranes resulted in an average density of 3.5 x 10 viable cells per ml. Ethanol production by reactors containing isotropic polypropylene fibers reached a maximum value of 26 g/liter-h based on the total reactor volume. Reactor performance depended on the fiber packing density and on the glucose medium flow rate and was limited by low nutrient and product transport rates. The inhibition of ethanol production and the reduction in fermentation efficiency arose primarily from the accumulation of CO(2) gas within the sealed reactor shell space. PMID- 16346347 TI - Bacteria and acidic drainage from coal refuse: inhibition by sodium lauryl sulfate and sodium benzoate. AB - The application of an aqueous solution of sodium lauryl sulfate and sodium benzoate to the surface of high-sulfur coal refuse resulted in the inhibition of iron-and sulfur-oxidizing chemoautotrophic bacteria and in the decrease of acidic drainage from the refuse, suggesting that acid drainage can be abated in the field by inhibiting iron- and sulfur-oxidizing bacteria. PMID- 16346349 TI - Ochratoxin A Production by Aspergillus ochraceus Wilhelm Grown Under Controlled Atmospheres. AB - [This corrects the article on p. 1136 in vol. 45.]. PMID- 16346348 TI - Isolation of cellulolytic actinomycetes from marine sediments. AB - The cellulolytic activity of 36 actinomycetes strains isolated from marine sediments was investigated by the cellulose-azure method. Approximately 50% of the isolates exhibited various degrees of cellulolytic activity. PMID- 16346350 TI - Microbial transformation of nitroaromatic compounds in sewage effluent. AB - [This corrects the article on p. 1239 in vol. 45.]. PMID- 16346351 TI - Inhibitory effect of solar radiation on amino Acid uptake in chesapeake bay bacteria. AB - The effect of solar radiation on a natural bacterial population from the Chesapeake Bay was evaluated from measured changes in numbers of organisms engaged in amino acid uptake. From July through May, freshly collected water samples were exposed in quartz containers to 3.5 h of total sunlight both with and without UV-absorbing filters. Water samples were subsequently incubated with tritiated amino acids, and the uptake-active bacteria were assayed by microauto radiography-epifluorescence microscopy. The survival index, defined as the fraction of the uptake-active population that remained active after the exposure to sunlight, ranged from 0.93 to 0.20. Decreased survival was correlated with increased solar intensity. The inhibition of amino acid uptake was attributed not only to the UV-B component of the solar spectrum (280 to 320 nm), but also to longer UV and visible wavelengths. PMID- 16346352 TI - Fungal biodegradation and biotransformation of soluble lignocarbohydrate complexes from straw. AB - Aspergillus japonicus is an efficient degrader of phenolics and carbohydrates present in a mixture of soluble lignocarbohydrate complexes extracted from wheat straw. Trichoderma sp. attacked part of the carbohydrate but hardly affected the aromatic portion of this solution. Polyporus versicolor had a complex effect; polymerization of low-molecular-size phenolics accompanied the degradation of aromatic and carbohydrate polymers. The addition of xylose to the medium facilitated depolymerization of lignin by the fungi tested and prevented the polymerization of low-molecular-size fractions of lignocarbohydrate complexes by P. versicolor. P. versicolor, in contrast to A. japonicus and Trichoderma sp., also excreted into the medium considerable amounts of laccase, but only in the absence of endogenous or exogenous carbohydrates. Apparently, laccase is involved in polymerization rather than degradation of lignin in this organism. A number of extracellular glycanases were also secreted by these fungi. PMID- 16346353 TI - Production of 3-hydroxypropionaldehyde from glycerol. AB - 3-Hydroxypropionaldehyde is a precursor to acrolein, which can be used as an intermediate for making acrylic acid and a variety of other useful industrial chemicals. Conversion of glycerol, a renewable resource, to 3 hydroxypropionaldehyde was attempted via action of glycerol dehydrase isolated from Lactobacillus sp. strain NRRL B-1720. This method, however, was unsatisfactory because enzyme activity was lost within 60 to 90 min after the reaction initiation. Fermentation of glycerol by whole cells of Klebsiella pneumoniae NRRL B-199 in the presence of optimal semicarbazide hydrochloride proved more effective. Using this technique, glycerol solutions of 30 g/liter yielded 3-hydroxypropionaldehyde solutions of 13.1 g/liter. Thus, a conversion efficiency equal to 55% of the theoretical maximum was realized. PMID- 16346354 TI - Effects of fermentation and its control on the sensory characteristics of conservolea variety green olives. AB - The various factors assumed to retard or even prevent the lactic acid fermentation of Conservolea variety green olives have been investigated. Of these, the sugar supplement along with the appropriate lactic starter culture supported a complete lactic acid fermentation. Acidification, particularly of starting brine, was shown to have little or no effect, whereas heat shock treatment improved the olive fermentability slightly without modifying the course of fermentation. These findings indicate that Conservolea olives do not contain phenolic compounds in concentrations inhibitory to lactic acid bacteria. PMID- 16346355 TI - Characterization of Gypsy Moth (Lymantria dispar) Nuclear Polyhedrosis Virus. AB - Characterization of the proteins and nucleic acid of the gypsy moth nuclear polyhedrosis virus isolated in Ithaca, N.Y. (LdNPV-IT) is presented. A total of 29 viral structural proteins were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis when the virus was isolated in the absence of alkaline protease activity. Fourteen surface envelope viral proteins were identified by lactoperoxidase iodination. Eleven proteins were associated with nucleocapsids prepared by Nonidet P-40 detergent treatment. Distinct alterations of viral proteins were documented when virions were purified in the presence of occlusion body-associated alkaline protease(s). Restriction enzyme digests of viral DNA indicated that this isolate was composed of a large number of genetic variants. On the basis of the major molar fragments resulting from EcoRI, BamHI, BglII, and HindIII digests, the molecular weight of the LdNPV genome was approximately 88 x 10. PMID- 16346356 TI - Conservation in soil of h(2) liberated from n(2) fixation by hup nodules. AB - Pigeon peas (Cajanus cajan) were grown in large soil columns (90-cm length by 30 cm diameter) and inoculated with four different strains of cowpea rhizobia, which varied with respect to hydrogen uptake activity (Hup). Despite the profuse liberation of H(2) from Hup nodules in vitro, H(2) gas was not detected in any of the soil columns. When H(2) was injected into the columns, the rates of consumption were highest in the treatments (including control) containing Hup nodules (218 and 177 nmol . h . cm) and lowest in the Hup treatments (158, 92, and 64 nmoles . h . cm). In situ H(2) uptake rates in small soil cores at fixed distances from the nodules decreased exponentially with distance from the nodule (R = 0.99). This decrease in H(2) consumption was associated with a similar decrease in numbers of H(2)-oxidizing chemolithotrophic bacteria as determined by the most-probable-number method. On the basis of two equations derived separately upon diffusive theory (Fix's Law) and kinetic theory (Michaelis-Menten), the empirically derived rate constants and coefficients indicated that all of the H(2) emitted from Hup nodules would be consumed by H(2)-oxidizing bacteria within a 3- to 4.5-cm radius of the nodule surface. It is concluded that H(2) is not lost from the soil-plant ecosystem during N(2) fixation in C. cajan but is conserved by H(2)-oxidizing bacteria. PMID- 16346357 TI - Isolation of a Pseudomonas sp. Which Utilizes the Phosphonate Herbicide Glyphosate. AB - A strain of bacteria has been isolated which rapidly and efficiently utilizes the herbicide glyphosate (N-phosphonomethylglycine) as its sole phosphorus source in a synthetic medium. The strain (PG2982) was isolated by subculturing Pseudomonas aeruginosa ATCC 9027 in a synthetic broth medium containing glyphosate as the sole phosphorus source. Strain PG2982 differs from the culture of P. aeruginosa in that it is nonflagellated, does not produce pyocyanin, and has an absolute requirement for thiamine. Strain PG2982 has been tentatively identified as a Pseudomonas sp. strain by its biochemical activities and moles percent guanine plus cytosine. Measurements of glyphosate with an amino acid analyzer show that glyphosate rapidly disappears from the medium during exponential growth of strain PG2982. In batch culture at 30 degrees C, this isolate completely utilized 1.0 mM glyphosate in 96 h and yielded a cell density equal to that obtained with 1.0 mM phosphate as the phosphorus source. However, a longer lag phase and greater generation time were noted in the glyphosate-containing medium. Strain PG2982 can efficiently utilize glyphosate as an alternate phosphorus source. PMID- 16346358 TI - Acidic Conditions Are Not Obligatory for Onset of Butanol Formation by Clostridium beijerinckii (Synonym, C. butylicum). AB - Factors that may initiate the metabolic transition for butanol production were investigated in batch cultures of Clostridium beijerinckii (synonym, Clostridium butylicum) VPI 13436. Cultures maintained at pH 6.8 produced nearly as much butanol as those incubated without pH control, indicating that neither a change in the culture pH nor acid conditions per se are always required to initiate solvent formation. Acetate and butyrate levels at the onset of butanol production were dependent on the pH at which the cultures were maintained. Cultures maintained at pH 6.8 could be accelerated into solvent production by artificially lowering the pH to 5.0 or by the addition of acetate plus butyrate without a pH change (but neither acid alone was effective). Solvent production was associated with slower rates of growth and general metabolism, and it did not show a requirement for mature spore formation. We speculate that a slowdown in metabolism, which may be brought about by several conditions, is mechanistically related to the onset of butanol production. Extracts of solvent-producing cells contained acetoacetate decarboxylase activity as well as higher NADP-linked butanol dehydrogenase and lower hydrogenase activities than extracts of acid producing cells. Solvent production did not appear to involve an enhanced ability to catalyze H(2) oxidation. PMID- 16346359 TI - Improved Microfouling Assay Employing a DNA-Specific Fluorochrome and Polystyrene as Substratum. AB - With a direct count assay, 10 fouling bacterial isolates have been characterized for their ability to adhere to glass cover slips and polystyrene dishes. Although most adhered in greater numbers to polystyrene, the preference was statistically significant for only seven isolates at the 95% confidence level, due in part to the greater variability in cell attachment to glass (coefficient of variation, 32.3% for glass compared with 10.0% for polystyrene). Employing polystyrene dishes, a novel microfouling assay was developed, based on the extraction and fluorometric determination of DNA. The assay was rapid, enabled the detection of as little as 0.15 mug of DNA per dish ( approximately 5,000 cells per mm), and showed good agreement with the direct count assay. The DNA method resulted in less variability among three replicates (average coefficient of variation, 7.06%) and allowed for estimation of bacterial density over a larger surface area per sample (1.89 x 10 mm) than was feasible with epifluorescence microscopy (0.06 to 0.1 mm). PMID- 16346360 TI - Enzyme-linked immunosorbent assays for detection of xylem-limited bacteria: use of trinder reagent. AB - Quantitation and detection of xylem-limited bacteria with an enzyme-linked immunosorbent assay with a peroxidase conjugate is described. The use of the Trinder reagent (4-aminoantipyrine) allows the determination of extremely small quantities of peroxidase with no precipitate formation or inactivation of the enzyme by H(2)O(2). Comparison of the enzyme-linked immunosorbent assay method with microscopic and histochemical tests for the presence of the phony peach disease bacterium in 9-year-old "June Gold" peach trees gave comparable results. The peroxidase conjugate with the Trinder reagent is more sensitive than the alkaline phosphatase conjugate typically used for enzyme-linked immunosorbent assay quantitation. PMID- 16346361 TI - Effect of H(2)-CO(2) on Methanogenesis from Acetate or Methanol in Methanosarcina spp. AB - Growth of Methanosarcina sp. strain 227 and Methanosarcina mazei on H(2)-CO(2) and mixtures of H(2)-CO(2) and acetate or methanol was examined. The growth yield of strain 227 on H(2)-CO(2) in complex medium was 8.4 mg/mmol of methane produced. Growth in defined medium was characteristically slower, and cell yields were proportionately lower. Labeling studies confirmed that CO(2) was rapidly reduced to CH(4) in the presence of H(2), and little acetate was used for methanogenesis until H(2) was exhausted. This resulted in a biphasic pattern of growth similar to that reported for strain 227 grown on methanol-acetate mixtures. Biphasic growth was not observed in cultures on mixtures of H(2)-CO(2) and methanol, and less methanol oxidation occurred in the presence of H(2). In M. mazei the aceticlastic reaction was also inhibited by the added H(2), but since the cultures did not immediately metabolize H(2), the duration of the inhibition was much longer. PMID- 16346362 TI - Quantitative high-pressure liquid chromatography-fluorescence determination of some important lower Fatty acids in lake sediments. AB - For the quantitative determination of traces of fatty acids in pore water, several gas and liquid chromatographic methods were tested and discussed. Direct determination by gas-liquid chromatography with the use of formic acid-saturated carrier gas was found to be the least laborious method, but it is only recommended for the determination of volatile acids such as acetate and higher homologs. For the determination of lactate and formate, a derivatization procedure is necessary. The determination of these acids as phenacyl or benzyl esters was complicated by contaminants in the reagents. For this reason, a high pressure liquid chromatography procedure with 4-bromomethyl-7-methoxycoumarin as a fluorescent labeling reagent is preferred. With this method, lactic, acetic, and formic acids could be demonstrated simultaneously at the nanogram level in 5 ml samples. Profiles of these acids in the sediment of Lake Vechten were measured, and they showed correlations with sulfate-reducing and methanogenic bacterial activities. PMID- 16346363 TI - Effect of methyl substitution on microbial degradation of linear styrene dimers by two soil bacteria. AB - The microbial degradation of 10 linear unsaturated dimers (I to IV) prepared from styrene and o-, m-, or p-methylstyrene was investigated with two soil bacteria, Alcaligenes sp. strain 559 and Pseudomonas sp. strain 419. The two strains decomposed styrene dimer I and all styrene-methylstyrene codimers II and III, but methylstyrene homodimers IV remained intact. The degradation rates of codimers II and III of o- and m-methylstyrenes were found to depend on both their structure and the strain used; i.e., Alcaligenes sp. strain 559 decomposed III faster than II, whereas the reverse order (II > III) was obtained with Pseudomonas sp. strain 419. In biodegradation by the former strain, the codimers were degraded faster in the presence of styrene dimer I than in its absence, but no such effect of dimer I was observed with the latter. PMID- 16346364 TI - Characterization of two extracellular polysaccharides from marine bacteria. AB - Two bacterial isolates from the intertidal zone produced significant quantities of extracellular polysaccharide with interesting properties. One polysaccharide was named PS 3a24; the other was named PS 3a35. The relative proportion of sugars in PS 3a35 was 51.6% glucose, 39.0% galactose, 3.1% mannose, and 6.3% rhamnose, with a trace of an unidentified sugar. PS 3a24 was composed of 40.2% glucose, 57.2% galactose, and 2.6% mannose. PS 3a35 contained 6% pyruvate, whereas PS 3a24 contained no pyruvate. Both exhibited high specific viscosity, pseudoplasticity, and stability over a wide range of pH in the presence of a variety of salts. The viscosity of PS 3a35 was relatively insensitive to increasing temperature, whereas that of PS 3a24 showed an irreversible drop on heating. PMID- 16346365 TI - Protection of sediment-adsorbed transforming DNA against enzymatic inactivation. AB - The action of DNase I on transforming DNA, both adsorbed to marine sediment and in solution, was investigated. DNase I reduced the transformation frequencies of free DNA more than of adsorbed DNA. Changes in salt concentration or pH did not have a significant influence on the DNA-sediment complex. Soil components other than organic materials and clay minerals can bind DNA and retard its enzymatic degradation. PMID- 16346366 TI - Purification and Characterization of alpha-Amylase from Bacillus licheniformis CUMC305. AB - alpha-Amylase produced by Bacillus licheniformis CUMC305 was purified 212-fold with a 42% yield through a series of four steps. The purified enzyme was homogeneous as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and discontinuous gel electrophoresis. The purified enzyme showed maximal activity at 90 degrees C and pH 9.0, and 91% of this activity remained at 100 degrees C. The enzyme retained 91, 79, and 71% maximal activity after 3 h of treatment at 60 degrees C, 3 h at 70 degrees C, and 90 min at 80 degrees C, respectively, in the absence of substrate. On the contrary, in the presence of substrate (soluble starch), the alpha-amylase enzyme was fully stable after a 4-h incubation at 100 degrees C. The enzyme showed 100% stability in the pH range 7 to 9; 95% stability at pH 10; and 84, 74, 68, and 50% stability at pH values of 6, 5, 4, and 3, respectively, after 18 h of treatment. The activation energy for this enzyme was calculated as 5.1 x 10 J/mol. The molecular weight was estimated to be 28,000 by sodium dodecyl sulfate-gel electrophoresis. The relative rates of hydrolysis of soluble starch, amylose, amylopectin, and glycogen were 1.27, 1.8, 1.94, and 2.28 mg/ml, respectively. V(max) values for hydrolysis of these substrates were calculated as 0.738, 1.08, 0.8, and 0.5 mg of maltose/ml per min, respectively. Of the cations, Na, Ca, and Mg, showed stimulatory effect, whereas Hg, Cu, Ni, Zn, Ag, Fe, Co, Cd, Al, and Mn were inhibitory. Of the anions, azide, F, SO(3), SO(4), S(2)O(3), MoO(4), and Wo(4) showed an excitant effect. p Chloromercuribenzoic acid and sodium iodoacetate were inhibitory, whereas cysteine, reduced glutathione, thiourea, beta-mercaptoethanol, and sodium glycerophosphate afforded protection to enzyme activity. alpha-Amylase was fairly resistant to EDTA treatment at 30 degrees C, but heating at 90 degrees C in presence of EDTA resulted in the complete loss of enzyme activity, which could be recovered partially by the addition of Cu and Fe but not by the addition of Ca or any other divalent ions. PMID- 16346367 TI - Some Properties of Thiosulfate-Oxidizing Enzyme from Marine Heterotroph 16B. AB - Thiosulfate-oxidizing enzyme has been demonstrated in cell-free extracts of the marine, thiosulfate-oxidizing pseudomonad strain 16B. The enzyme, partially purified by ion-exchange chromatography and calcium phosphate gel treatment, catalyzed the oxidation of thiosulfate to tetrathionate with the concomitant reduction of ferricyanide. Native but not mammalian cytochrome c was also reduced by the enzyme in the presence of thiosulfate. The enzyme was located exclusively in the supernatant of ultracentrifuged cell extracts. The most purified enzyme preparation, like intact cells, exhibited a temperature optimum of 30 to 31 degrees C. However, it exhibited no definite pH optimum. At pH 6.1 to 6.3 and 30 degrees C, the K(m) for thiosulfate was 1.57 mM. At lower temperatures, the apparent K(m) for thiosulfate increased, but the apparent maximum velocity remained virtually unchanged. Thiosulfate oxidation in intact cells exhibited an increase in the pH optimum at lower temperatures. The thiosulfate-oxidizing enzyme of marine heterotroph 16B is compared with thiosulfate-oxidizing enzymes from other bacteria, and the effect of temperature on the relationship between pH and thiosulfate oxidation is discussed with reference to the natural habitat of the bacterium. PMID- 16346368 TI - Inexpensive flow microcalorimeter for measuring heat production of attached and sedimentary aquatic microorganisms. AB - A relatively inexpensive flow microcalorimeter is described which is capable of detecting heat outputs as low as 3 muW (precision, +/-2%). Its use is illustrated on river epilithon (0.8 to 6.8 muW cm), river sand (9.8 muW cm), and marine sand (15.3 muW cm); however, it could be used to detect the heat output from any biotic material over which a flow of water can be passed, provided that such an action would not be disruptive to chemical and biological equilibria. PMID- 16346369 TI - Novel Biotransformations of 7-Ethoxycoumarin by Streptomyces griseus. AB - Biotransformation of 7-ethoxycoumarin by Streptomyces griseus resulted in the accumulation of two metabolites which were isolated and identified as 7 hydroxycoumarin and 7-hydroxy-6-methoxycoumarin. A novel series of biotransformation reactions is implicated in the conversion of the ethoxycoumarin substrate to these products, including O-deethylation, 6-hydroxylation to form a 6,7-dihydroxycoumarin catechol, and subsequent O-methylation. Either 7 hydroxycoumarin or 6,7-dihydroxycoumarin was biotransformed to 7-hydroxy-6 methoxycoumarin by S. griseus. Trace amounts of the isomeric 6-hydroxy-7 methoxycoumarin were detected when 6,7-dihydroxycoumarin was used as the substrate. Efforts to obtain a cell-free catechol-O-methyltransferase enzyme system from S. griseus were unsuccessful. However, [methyl-C]methionine was used with cultures of S. griseus to form 7-hydroxy-6-[C]methoxycoumarin. PMID- 16346370 TI - Microbial transformation of macrocyclic trichothecenes. AB - A resting culture of Rhizopus arrhizus (ATCC 11145) transformed verrucarin A into 16-hydroxyverrucarin A, whereas R. arrhizus transformed verrucarin B into a mixture of 16-hydroxyverrucarin B and 3'-hydroxyverrucarin A. Relative to verrucarins A and B, the 16-hydroxy derivatives showed marked increases in activity, as tested in vivo against P388 mouse leukemia. PMID- 16346371 TI - Isolation and Characterization of Methanomicrobium paynteri sp. nov., a Mesophilic Methanogen Isolated from Marine Sediments. AB - A new mesophilic methanogenic bacterial species isolated from marine sediments collected in the Cayman Islands is described. Cells are small rods occuring singly without filaments, are not motile, and do not possess flagella. Colonies are semitransparent and off-white in color. After 2 weeks of incubation at 37 degrees C colonies are 1 to 2 mm in size, circular, and have entire edges. Only hydrogen-carbon dioxide is a substrate for growth and methane formation. Cells can tolerate a variety of organic secondary buffers (bicarbonate-CO(2) being the primary buffer). Cells do not require yeast extract or Trypticase, but do require acetate, for growth. The optimum growth temperature is 40 degrees C. The optimum sodium concentration is 0.15 M. The optimum pH for growth is 7.0. The minimum generation time is 4.8 h. The DNA base composition is 44.9 mol% guanine plus cytosine. The name Methanomicrobium paynteri is proposed in honor of M. J. B. Paynter. The type strain is G-2000 (=ATCC 33997, =DSM 2545). PMID- 16346372 TI - Technique for enumeration of heterotrophic and phototrophic nanoplankton, using epifluorescence microscopy, and comparison with other procedures. AB - A new method is described that uses the fluorochrome primulin and epifluorescence microscopy for the enumeration of heterotrophic and phototrophic nanoplankton (2 to 20 mum). Phototrophic microorganisms are distinguished from heterotrophs by the red autofluorescence of chlorophyll a. Separate filter sets are used which allow visualization of the primulin-stained nanoplankton without masking chlorophyll a fluorescence, thus allowing easy recognition of phototrophic cells. Comparison with existing epifluorescence techniques for counting heterotrophic and phototrophic nanoplankton shows that primulin provides more accurate counts of these populations than the fluorescein isothiocyanate or proflavine techniques. Accuracy is comparable to that with the acridine orange technique, but this method requires only one filter preparation for the enumeration of both phototrophic and heterotrophic populations. PMID- 16346373 TI - Weighing technique for determining bacterial dry mass based on rate of moisture uptake. AB - Weights were recorded at 30-s intervals timed from initial exposure of dried cells to atmosphere. Linear regressions of data over 3 min gave moisture-free dry mass estimates. PMID- 16346374 TI - Improved method for isolation of bacterial inhibitors from oleuropein hydrolysis. AB - A new high-pressure liquid chromatography multidetection quantitative method for the isolation of the products of oleuropein hydrolysis is described. A single analysis yields sufficient amounts of the compounds to test their inhibitory effect on bacterial growth. PMID- 16346375 TI - Isolation and Characterization of a Xylanase from Bacillus subtilis. AB - Partial characterization of an extracellular xylanase isolated by chromatography from Bacillus subtilis gave a molecular weight of 32,000 and optimum pH and temperature of 5.0 and 50 degrees C, respectively. K(m) and V(max) values, determined with a soluble larchwood xylan, were 0.16% and 7.0 x 10 mumol min mg of enzyme respectively. The amino acid composition showed more basic amino acid residues than in a previously characterized xylanase from a white-rot fungus. PMID- 16346376 TI - Evaluation of Nitrate Reductase Activity in Rhizobium japonicum. AB - Nitrate reductase activity was evaluated by four approaches, using four strains of Rhizobium japonicum and 11 chlorate-resistant mutants of the four strains. It was concluded that in vitro assays with bacteria or bacteroids provide the most simple and reliable assessment of the presence or absence of nitrate reductase. Nitrite reductase activity with methyl viologen and dithionite was found, but the enzyme activity does not confound the assay of nitrate reductase. PMID- 16346377 TI - Isolation, Purification, and Some Properties of Penicillium chrysogenum Tannase. AB - Tannase isolated from Penicillium chrysogenum was purified 24-fold with 18.5% recovery after ammonium sulfate precipitation, DEAE-cellulose column chromatography, and Sephadex G-200 gel filtration. Optimum enzyme activity was recorded at pH 5.0 to 6.0 and at 30 to 40 degrees C. The enzyme was stable up to 30 degrees C and within the pH range of 4.0 to 6.5. The K(m) value was found to be 0.48 x 10 M when tannic acid was used as the substrate. Metal salts at 20 mM inhibited the enzyme to different levels. PMID- 16346379 TI - Production and Characteristics of Raw Starch-Digesting Glucoamylase O from a Protease-Negative, Glycosidase-Negative Aspergillus awamori var. kawachi Mutant. AB - [This corrects the article on p. 906 in vol. 45.]. PMID- 16346378 TI - Sampling strategy for detecting viruses in a sewage treatment plant. AB - [This corrects the article on p. 1769 in vol. 45.]. PMID- 16346380 TI - Fermentative Degradation of Polyethylene Glycol by a Strictly Anaerobic, Gram Negative, Nonsporeforming Bacterium, Pelobacter venetianus sp. nov. AB - [This corrects the article on p. 1913 in vol. 45.]. PMID- 16346381 TI - Sensitivity of an oligotrophic lake planktonic bacterial community to oxygen stress. AB - Dissolved oxygen at approximately four times normal saturation (42 mg liter) inhibited the growth and metabolism of summer planktonic bacteria in the surface water of alpine oligotrophic Mountain Lake (Giles County, Va.). Data were derived from growth of CFU on membrane filters, d-[U-C]glucose incorporation into the extractable lipid of these CFU, and respiration and assimilation of d-[U C]glucose by lake water samples. Statistically significant (alpha < 0.05) differences were not detected in either CFU or C incorporation in lipid when superoxide dismutase (30 U ml) or catalase (130 U ml) was added to the medium. Thus, exogenous oxygen by-products apparently are not responsible for the observed inhibition of growth and metabolism. PMID- 16346382 TI - Identification and enumeration of marine chroococcoid cyanobacteria by immunofluorescence. AB - We used an indirect immunofluorescence technique to permit the identification and enumeration of individual or closely related strains of chroococcoid cyanobacteria of the general Synechococcus and Synechocystis in natural seawater samples. Antisera directed against each of five strains (two phycoerythrin containing Synechococcus strains, two phycocyanin-containing Synechococcus strains, and one Synechocystis strain) were produced and tested for cross reactions with cultures of a variety of cyanobacteria and representatives of other algae and bacteria. Each antiserum was relatively specific. The observed cross-reactions occurred between strains that were isolated from similar oceanic environments. We were able, therefore, to apply this technique to field samples. Preliminary results for April to December 1982 in Great South Bay, New York, show that Synechocystis populations are present only during spring and summer, phycocyanin-containing Synechococcus strains are only a minor component in the spring and summer, and phycoerythrin-containing Synechococcus populations become significant in summer and remain so until late fall or winter. PMID- 16346383 TI - Wide-host-range plasmids function in the genus thiobacillus. AB - Plasmids S-a, RP4, R388, and several RP4 derivatives (pMD101, pDT387, and pDT566) were transmissible by conjugation to Thiobacillus novellus from Escherichia coli. Genetic markers were expressed in T. novellus, with the exception of chloramphenicol resistance and ampicillin resistance. Plasmids were not transmissible by conjugation from E. coli donors to Thiobacillus intermedius, T. perometabolis, T. neapolitanus, or T. acidophilus recipients, although they could be mated into these strains from T. novellus. All Thiobacillus species tested could transfer plasmids back to E. coli, with the exception of T. acidophilus. The donor-specific bacteriophages PRR1 and PRD1 were incapable of initiating the lytic cycle in RP4-bearing strains of T. novellus. The cosmid cloning vehicle pVK100 could be mobilized from E. coli to T. novellus with the aid of the "helper" plasmid pRK2013. pVK100 is stable in T. novellus, but pRK2013 is not maintained in this species. pRK2013 was also used to mobilize another cloning vector, R300B, to T. novellus. A previously unreported cryptic plasmid of approximately 24 megadaltons was observed in T. intermedius. No native plasmids were demonstrated in the other Thiobacillus species except in T. acidophilus, which contained cryptic plasmids ranging in size from 7.6 to 56 megadaltons (molecular mass). PMID- 16346384 TI - Highly Efficient Mutagenesis of Claviceps purpurea by Using Protoplasts. AB - Claviceps purpurea ATCC 20102, which is aconidial under laboratory conditions, was grown in submerged culture in the presence of mutagens and various nutritional additives. Protoplasts from such cultures were prepared and regenerated on solid medium to obtain colonies from single cell units. Frequencies of auxotrophs and high alkaloid producers were on the order of 1 to 2%. Some of the auxotrophic mutants derived from strain ATCC 20102 were constantly segregating prototrophs. High-alkaloid-producing derivatives showed sclerotia-like morphology and violet-brown pigmentation, in contrast to the parent strain; some of them also showed segregation sectors when grown as giant colonies. Mutagenesis of strain 1029, isolated during this study and having an increased level of alkaloid synthesis and sclerotia-like cell morphology, was done in the same fashion as with the original parent strain, ATCC 20102. Mutants obtained from this strain were all stable with respect to their genotypes. However, a large proportion of colonies derived from regenerated protoplasts, even in the mutagen-free controls, showed a lowered level of alkaloid production and were morphologically more similar to the original wild type, ATCC 20102. The influence of protoplast preparation or regeneration or both on the stability of genes involved in differentiation is discussed. PMID- 16346385 TI - Cell Wall Composition of Neurospora crassa Under Conditions of Copper Toxicity. AB - The mycelia of Neurospora crassa grown in the presence of high concentrations of copper were blue in color, but only on a medium containing inorganic nitrate and phosphate as the nitrogen and phosphate sources, respectively. The cell wall isolate of the blue mycelia contained large amounts (12%) of copper and higher amounts of chitosan, phosphate, and amino groups, with a 42% decrease in the chitin content. Although all the glucosamine of the cell wall of control cultures could be released within 6 h of hydrolysis with acid, that of the blue mycelium required prolonged hydrolysis for 24 h. On removal of copper, the cell wall of the blue mycelium could quantitatively bind again to copper as well as to zinc. Although zinc binding was fivefold greater, copper alone was preferentially bound from a mixture of the two metal ions. Supplementation of iron along with copper in the culture medium resulted in the disappearance of the blue color of the mycelium and restoration of normal growth and composition of the cell wall, probably by limiting the uptake of copper from the medium. The possibility of the cell wall being a specific site of lesion in copper toxicity in the mold is discussed. PMID- 16346386 TI - N(2) Fixation by Azospirillum brasilense and Its Incorporation into Host Setaria italica. AB - Growth and nitrogen fixation were followed during the life cycle of Setaria italica (foxtail millet) inoculated with Azospirillum brasilense in controlled environment growth chambers. The plants were fertilized at seeding with a limiting amount of combined nitrogen and maintained with an N-free mineral solution. During maturation of the plants, substantial nitrogenase activity, measured by acetylene reduction, developed in the rhizosphere, with total fixation estimated to be equivalent to 20% of the N in the inoculated plants. The peak of this activity coincided with depletion of soluble nitrogen from the system, which in turn was reflected by a sharp decrease in the nitrate reductase activity of the leaves. A. brasilense was found in association with the root populations at 8 x 10 cells per gram of dry weight. An increase in shoot growth occurred at this time, but no significant increase in total plant nitrogen could be demonstrated. N(2) enrichment experiments confirmed that fixation was occurring, but only about 5% of the nitrogen fixed by A. brasilense was incorporated into the plants within 3 weeks. There was thus no evidence of direct bacterium-to-plant transport of fixed nitrogen, but rather a slow transfer suggesting the gradual death of bacteria and subsequent mineralization of their nitrogen, at least under growth-room conditions. PMID- 16346387 TI - Analysis of Distributed Growth of Saccharomyces cerevisiae Cells Immobilized in Polyacrylamide Gel. AB - A technique is described for the quantitative determination of the distributed growth of Saccharomyces cerevisiae immobilized in polyacrylamide gel. Gel specimens were embedded in paraffin or gelatin and paraffin before sectioning and staining. Photomicrographs of specimen sections were enlarged, and cell microcolony volumes were determined as a function of position in the gel by grid transparency analysis. Overall cell densities within the gel were calculated for a quantitative comparison with values measured by a second spectrophotometric method. The results show good agreement and demonstrate the sigmoidal growth of the immobilized cells, reaching a maximum steady-state value. The technique shows promise as a general method for following the transient growth of organisms immobilized within gel particles. PMID- 16346388 TI - Mesophilic cellulolytic clostridia from freshwater environments. AB - Eight strains of obligately anaerobic, mesophilic, cellulolytic bacteria were isolated from mud of freshwater environments. The isolates (C strains) were rod shaped, gram negative, and formed terminal spherical to oval spores that swelled the sporangium. The guanine plus cytosine content of the DNA of the C strains ranged from 30.7 to 33.2 mol% (midpoint of thermal denaturation). The C strains fermented cellulose with formation primarily of acetate, ethanol, CO(2), and H(2). Reducing sugars accumulated in the supernatant fluid of cultures which initially contained >/=0.4% (wt/vol) cellulose. The C strains resembled Clostridium cellobioparum in some phenotypic characteristics and Clostridium papyrosolvens in others, but they were not identical to either of these species. The C strains differed from thermophilic cellulolytic clostridia (e.g., Clostridium thermocellum) not only in growth temperature range but also because they fermented xylan and five-carbon products of plant polysaccharide hydrolysis such as d-xylose and l-arabinose. At 40 degrees C, cellulose was degraded by cellulolytic mesophilic cells (strain C7) at a rate comparable to that at which C. thermocellum degrades cellulose at 60 degrees C. Substrate utilization and growth temperature data indicated that the C strains contribute to the anaerobic breakdown of plant polymers in the environments they inhabit. PMID- 16346389 TI - Conjugal Transfer of Broad-Host-Range Plasmid pAMbeta1 into Enteric Species of Lactic Acid Bacteria. AB - The broad-host-range plasmid pAMbeta1, which codes for erythromycin and lincomycin resistance, was transferred by conjugation into Lactobacillus acidophilus, Lactobacillus reuteri, and Lactobacillus salivarius. A novel 17 megadalton plasmid molecule was detected in the transconjugants, confirming the introduction of pAMbeta1 into each species. PMID- 16346390 TI - Fungal toxicity of mobilized soil aluminum and manganese. AB - Inhibition of Aspergillus flavus growth from spores was used as a simple bioassay for toxicity of Al and Mn mobilized by simulated acid precipitation. Al was identified as being toxic in soil leachates resulting from acid inputs of pH less than 2.7. Inhibition by Mn was not detectable. The addition of fluoride significantly reduced Al toxicity, suggesting that biotoxicity of Al is partially dependent on the anionic composition of the soil solution. PMID- 16346391 TI - Identification of Low-Molecular-Weight Nucleic Acid-Related Substances Secreted by Streptomyces aureofaciens. AB - Streptomyces aureofaciens growth in chemically defined medium is actively associated with the secretion of low-molecular-weight nucleic acid-related substances and is linked to low availability of phosphate. Thirteen pure compounds were isolated, of which seven were identified. PMID- 16346392 TI - Effects of pentachloronitrobenzene and some of its known and possible metabolites on fungi. AB - Fungicidal activities of pentachloronitrobenzene and derivatives were tested with Candida albicans, Aspergillus fumigatus, Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporon canis. In relation to pentachloronitrobenzene, no increasing fungistatic activities were found with cysteine derivatives. Rising fungistatic activities were seen with pentachlorophenylmethylsulfone and, in some of the strains, with pentachloronitrosobenzene, pentachlorothiophenol, pentachlorophenol, pentachloroaniline, pentachlorophenylmethylsulfoxide, the isomeric tetrachloronitrobenzenes, tetrachlorothiophenols, tetrachlorophenols, and tetrachloroanilines. PMID- 16346393 TI - Controlling Morphological Instability of Zymomonas mobilis Strains in Continuous Culture. AB - [This corrects the article on p. 1900 in vol. 45.]. PMID- 16346395 TI - Effect of microorganisms on in situ uranium mining. AB - The extraction of some metal values, e.g., uranium or copper, may be accomplished by using solutions to remove metals from ore bodies without practicing conventional mining. This process is referred to as in situ leaching and has been used industrially to recover uranium. The growth of microbial populations during in situ leaching is believed to be one of the causes of flow path plugging in the ore body, which results in decreased uranium production. Leach solution and solid samples from well casings and submersible pumps were collected from an in situ mining operation experiencing plugging problems. Bacillus sp., Micrococcus sp., pseudomonads, and xanthomonads were isolated from these samples in concentrations of 10 CFU ml. A mixed culture of these organisms was inoculated into a uranium core specimen in the laboratory to assess the role of microbes in the plugging problem. A one-third decrease in permeability was effected in 16 days. Hydrogen peroxide (0.2 g liter) killed the microorganisms in the core and alleviated the plugging problem. Periodically injecting hydrogen peroxide into the ore body through the production wells may reduce microbial plugging problems. PMID- 16346394 TI - Freeze-Recovery Physiology of Nitrogenase Activity in Terrestrial Nostoc sp. Colonies. AB - Nostoc sp. colonies from field collections were cultured and propagated on silica sand with aqueous N-free BG-11 medium. Laboratory experiments were conducted to characterize the in vivo freeze-recovery physiology of nitrogenase activity. Nitrogenase activity was monitored by the acetylene reduction technique. Frozen Nostoc sp. colonies were thawed and warmed to 10, 15, 20, 25, or 30 degrees C. At 25 and 30 degrees C, nitrogenase activity was detected within 6 h after thawing. At 20 degrees C or lower, nitrogenase activity was not detected until 12 h after thawing. Optimum thawing temperature with respect to the recovery of nitrogenase activity was 25 degrees C. In subsequent experiments, laboratory-grown Nostoc colonies were used along with the following conditions: prefreezing treatment of 3 days of exposure to light or darkness, freezing, and then thawing to 25 degrees C in light or darkness with or without metabolic inhibitors [3-(3,4 dichlorophenyl)-1, 1-dimethylurea (DCMU), monofluoroacetate, or chloramphenicol]. Approximately 30% of the energy in the initial recovery of nitrogenase activity (to 12 h after thawing) appeared to be supplied via the utilization of carbon compounds stored before freezing. Photosynthetic conditions (i.e., light and without DCMU) were necessary for maximum recovery of nitrogenase activity. In the presence of the protein synthesis inhibitor chloramphenicol, nitrogenase activity was still detected at 12 to 48 h after thawing. Although damage may occur to nitrogenase, some of the enzyme was capable of surviving the freeze-thaw period in vivo. However, complete recovery of nitrogenase activity (equal to prefreezing activity) may entail some de novo synthesis of nitrogenase. PMID- 16346397 TI - Role of tween 80 and monoolein in a lipid-sterol-protein complex which enhances ethanol tolerance of sake yeasts. AB - An exogenous ternary complex composed of Tween 80, ergosterol, and albumin increased the final ethanol concentration of fermentation by sake yeasts from 17.2 to 19.0% (vol/vol) and reduced the fermentation time from 30 to 25 days. Likewise, a complex of monoolein, albumin, and either ergosterol or ergosteryl oleate increased the final ethanol concentration of fermentation to 19.7 or 19.8% (vol/vol), respectively, and reduced the fermentation time to 25 days. Both Tween 80 and monoolein promoted the fermentative activity (Q(CO(2) )) of cells, and the effect was enhanced by the presence of ergosterol. PMID- 16346396 TI - Influence of Calcium and Manganese on Dechaining of Lactobacillus bulgaricus. AB - The events responsible for the transition of Lactobacillus bulgaricus 1243-F from long filamentous chains to short bacilloid rods were examined in a cation depleted liquid medium. In the presence of magnesium only, cells grew as long chains of unseparated cells. The addition of 100 muM to 1 mM calcium or manganese to this medium resulted in the dechaining of these cells to short bacilloid rods. Fe, Zn, Co, and Cu failed to induce dechaining. Induction of calcium and manganese dechaining functioned under controlled pH maintained at 5.0 and 6.0 but not at pH 7.0. This was consistent with a previous report showing failure in synthesis of dechaining enzymes by L. bulgaricus under pH conditions approaching alkalinity (S. K. Rhee and M. Y. Pack, J. Bacteriol. 144:865-868, 1980). We conclude that under pH conditions which permit synthesis of dechaining enzymes, calcium and manganese are necessary for dechaining activity. PMID- 16346398 TI - GELRITE as an Agar Substitute in Bacteriological Media. AB - GELRITE gellan gum (formerly known as PS-60 and S-60) is a new naturally derived, highly purified polysaccharide which displays several interesting properties, including selfgelling. The suitability of GELRITE as an agar substitute was tested by evaluating the performance of several media selected from among those most commonly used in the isolation, identification, and enumeration of microorganisms in clinical laboratories. Fifty different bacterial species previously implicated in human infections served as test strains. On the basis of the various parameters considered, namely, colony characteristics, biochemical reactions, hemolytic patterns, and plating efficiency, media gelled by agar and by GELRITE compared quite favorably. PMID- 16346399 TI - Medium Promoting Sporulation of Bacillus larvae and Metabolism of Medium Components. AB - A new medium, designated TMYGP broth, was developed that allowed the honeybee pathogen Bacillus larvae NRRL B-3650 to produce up to 5 x 10 spores per ml of culture (microscopic count). This species normally sporulates poorly, if at all, in artificial broth media. An aeration rate lower than that normally used to cultivate other Bacillus species was required for sporulation. During the exponential growth phase, acids were produced by catabolism of yeast extract components, causing a decrease in pH of the medium. Thereafter, the pH began to increase, probably because of derepression of the citric acid cycle and consumption of the acids. Only after this time did usage of glucose from the medium occur. Thus, glucose usage seems to be regulated by catabolite repression. The presence of glucose was needed for one or more of the later events of sporulation. Of many substances tested, only gluconic acid and glucosamine partially substituted for glucose as a requirement for sporulation. Pyruvate was also required for good sporulation. It was metabolized during the late exponential phase of growth. PMID- 16346400 TI - Competition of Rhizobium japonicum Strains in Early Stages of Soybean Nodulation. AB - The effects of preexposure of soybean (Glycine max L. Merrill) roots to Rhizobium japonicum strains and subsequent establishment of other strains in the nodules were investigated by using combinations of effective strains (USDA 110 and USDA 138) and effective-ineffective strains (USDA 110 and SM-5). Strain USDA 110 was a better competitor than either USDA 138 or SM-5 on cultivars Lee and Peking. However, when either of the two less-competitive strains was inoculated into 2 day-old seedlings before USDA 110 was, their nodule occupancy increased significantly on both cultivars. With USDA 138 as the primary inoculum and USDA 110 delayed for 6, 48, and 168 h, the incidence of USDA 138 nodules increased on cultivar Peking from 6% (at zero time) to 28, 70, and 82% and on cultivar Lee from 17% (at zero time) to 32, 88, and 95% for the three time delays, respectively. Preexposure of 2-week-old roots of cultivar Lee to USDA 138 had essentially the same effect: the incidence of USDA 138 nodules increased from 23% at zero time to 89 and 97% when USDA 110 was delayed for 24 and 72 h, respectively. When the ineffective strain SM-5 was used as the primary inoculum, followed by USDA 110 72 h later, the percentage of nodules containing SM-5 increased from 7 to 76%. These results indicate that the early events in the nodulation process of soybeans are perhaps the most critical for competition among R. japonicum strains. PMID- 16346401 TI - Occurrence of lactic Acid bacteria during the different stages of vinification and conservation of wines. AB - We showed that the growth of lactic acid bacteria during alcoholic fermentation depends on the composition of the must. We illustrated how the addition of sulfur dioxide to the must before fermentation and the temperature of storage both affect the growth of these bacteria in the wine. Whereas species of Lactobacillus and Leuconostoc mesenteroides were isolated from grapes and must, Leuconostoc oenos was the only species isolated after alcoholic fermentation. This organism was responsible for the malolactic fermentation. Isolates of this species varied in their ability to ferment pentoses and hexoses. The survival of Leuconostoc oenos in wines after malolactic fermentation depended on wine pH, alcohol concentration, SO(2) concentration, and temperature of storage. PMID- 16346402 TI - Interaction of Agromyces ramosus with Other Bacteria in Soil. AB - Agromyces ramosus occurs in very high numbers in most soils and, based on studies of laboratory isolates, does not require host cells for growth. Nevertheless, it attacked and destroyed most of the gram-positive and gram-negative bacterial species tested as possible host organisms. A. ramosus also attacked and destroyed Saccharomyces cerevisiae. The possibility of attack on fungi was unclear. Among the bacteria serving as hosts were the important soil species Azotobacter vinelandii, Rhizobium leguminosarum, Rhizobium meliloti, and Agrobacterium tumefaciens. Dead cells were not attacked. A. vinelandii cysts were attacked but left unharmed. To some extent, A. vinelandii seemed to survive this attack by encysting. Attack by A. ramosus occurred in natural soil and over a broad range of nutritional levels in laboratory media. The attack did not seem to be a means for obtaining an increased supply of commonly available nutrients. Instead, it seemed to be a means of obtaining something produced, perhaps in small amounts, by a variety of organisms, but not by all organisms. Several types of culture filtrates were tested for activity. The filtrates neither stimulated nor inhibited the growth of A. ramosus or the host organisms. The availability of catalase activity in host organisms did not seem to be involved. It is not known whether the attack by Agromyces ramosus in soil can be manipulated to cause a decrease in numbers of Agrobacterium tumefaciens or other pathogens without simultaneously depressing the numbers of beneficial organisms in this habitat. PMID- 16346403 TI - Possible involvement of a megaplasmid in nodulation of soybeans by fast-growing rhizobia from china. AB - Several isolates from a newly described group of fast-growing acid-producing soybean rhizobia, Rhizobium japonicum, were analyzed for plasmid content. All contained from one to four plasmids with molecular weights of 100 x 10 or larger. Although most of the isolates shared plasmids of similar size, the restriction endonuclease (BamHI, EcoRI, and HindIII) patterns of the plasmids from three of the isolates were vastly different. Growth in the presence of acridine orange was effective in producing mutants cured of the largest plasmid in one of the strains. These mutants also lost the ability to form nodules on soybeans. High temperature curing of a smaller plasmid in another strain did not lead to loss of nodulating ability or alteration of symbiotic effectiveness on soybean cultivars. The identities of all of the isolates and mutants were ascertained by immunofluoresence and immunodiffusion. The new fast-growing strains of R. japonicum may provide a better genetic system for the study of the soybean symbiosis than the slow-growing R. japonicum, not all of which can be shown to contain plasmids. PMID- 16346404 TI - Microbial Production of Optically Pure l-Iditol by Yeast Strains. AB - Several yeast strains belonging to genus Candida were found to selectively hydrogenate l-sorbose with enantiomeric specificity, yielding optically pure l iditol in the culture broth. The most active strain, isolated from a commercial lemon, was identified as Candida intermedia, which produced 50 g of l-iditol per liter from 150 g of l-sorbose per liter during a 5-day fermentation period (35% yield). PMID- 16346405 TI - Effect of UV Light on Spore Viability and Mosquito Larvicidal Activity of Bacillus sphaericus 1593. AB - UV light from a germicidal lamp rapidly reduced the viability of Bacillus sphaericus 1593 spores, but insecticidal activity was resistant to inactivation by continuous exposure to UV light for 4 h. PMID- 16346406 TI - Effect of storm events on benthic nitrifying activity. AB - Storm events resulted in a marked reduction in the benthic nitrifying activity of a stream receiving geothermal inputs of ammonium. Subsequently, nitrifying activity demonstrated a logarithmic increase until a stable activity was reached 10 to 12 days after the storm event. The rate of increase of poststorm nitrifying activity was used to calculate the nitrifier growth rate in situ (0.0075 to 0.0116 h) which was lower than the growth rates observed for laboratory cultures of nitrifying bacteria isolated from the sediments (0.0233 to 0.0990 h). PMID- 16346407 TI - Nonlinear estimation of monod growth kinetic parameters from a single substrate depletion curve. AB - [This corrects the article on p. 1454 in vol. 45.]. PMID- 16346408 TI - Predicting 2,4-dichlorophenoxyacetic Acid ester transformation rates in periphyton-dominated ecosystems. AB - [This corrects the article on p. 149 in vol. 46.]. PMID- 16346409 TI - Ecology and Physiology of the Competition for Light Between Chlorobium limicola and Chlorobium phaeobacteroides in Natural Habitats. AB - Accurate studies of the pigment composition and isolation in pure cultures of Chlorobiaceae from samples of eight Spanish lakes show that there are two main coexisting groups of green and brown Chlorobium spp. represented respectively by Chlorobium limicola and Chlorobium phaeobacteroides. Laboratory experiments with pure and mixed cultures of the isolated strains show that light quality plays a selective role on the species composition among Chlorobiaceae. This selection depends on the pigment composition which determines the in vivo absorption spectrum of the cells as well as on their ability to adjust the intracellular concentration of light-harvesting pigments to the spectral distribution and energy of light. Correlation analysis performed with field data resulted in significant, but low, correlation coefficients. Nevertheless, they were consistent with laboratory data showing that brown Chlorobiaceae were dominant in deep layers in meromictic lakes, whereas green Chlorobiaceae dominated in layers nearer the surface or underneath plates of Chromatiaceae. The combination of laboratory and field observations stress the role of biological light filtering in determining the species composition among Chlorobiaceae in lakes. PMID- 16346410 TI - System development for linked-fermentation production of solvents from algal biomass. AB - Five species of the genus Dunaliella (D. tertiolecta, D. primolecta, D. parva, D. bardawil, and D. salina) were examined for glycerol accumulation, growth rate, cell density, and protein and chlorophyll content. The suitability of each algal species for use as a fermentation substrate was judged according to glycerol accumulation and quantities of neutral solvents produced after sequential bacterial fermentations. When grown in 2 M NaCl, with 24 mM NaHCO(3) or 3% CO(2) at 28 degrees C and with 10,000 to 15,000 lx of incident light on two sides of a glass aquarium, four of the five species tested produced ca. 10 to 20 mg of glycerol per liter of culture. Clostridium pasteurianum was found to convert an algal biomass mixture supplemented with 4% glycerol to ca. 16 g of mixed solvents (n-butanol, 1,3-propanediol, and ethanol) per liter. Acetone was not detected. Additionally, it has been demonstrated that Dunaliella concentrates of up to 300 fold can be directly fermented to an identical pattern of mixed solvents. Overall solvent yields were reduced by >50% when fermentations were performed in the presence of 2% NaCl. These results are discussed in terms of practical application in tropical coastal zones. PMID- 16346411 TI - Long-Term Changes in Chemostat Cultures of Cytophaga johnsonae. AB - Long-term studies with a gliding, heterotrophic bacterium, Cytophaga johnsonae, were conducted in a glucose-limited chemostat at a high and a low dilution rate. To test the stability of the steady state during long-term experiments the following parameters were monitored: optical density, glucose concentration, glucose uptake potential, ATP content of the cells, and plate counts on two different agar media. Biomass remained relatively constant, although the observed changes could have been possible in both directions. During all steady states, glucose uptake showed a stepwise increase and the glucose concentration showed a corresponding decrease. Glucose uptake potential and glucose concentration in the chemostat were inversely proportional. The ATP content of the cells varied up to 33% during the steady state, but did not show a general trend. After long cultivation in all chemostats, plate counts on both agars dropped to values less than 20% of the original steady-state level. These decreases were due to an inability of the cells to grow on agar plates, not to a lack of vitality of the cells in the chemostat. This study showed that even during shorter chemostat runs, e.g., 1 week, changes in important parameters with the steady state must be expected, especially in the uptake potential and the concentration of the limiting substrate. PMID- 16346412 TI - Immunological Characterization of Pectinatus cerevisiophilus Strains. AB - Eleven Pectinatus cerevisiophilus strains of brewery origin were classified serologically by gel diffusion precipitin tests, immunoelectrophoresis, and the fluorescent antibody staining technique. The Pectinatus strains could be assigned immunologically to three different groups. Groups I and III were found to be very closely related, and only some of the antisera used showed differences. The antisera against the strains belonging to group II contained a common group antigen. A strong precipitation band found near the antigen was shown to represent the interaction of the lipopolysaccharide antibody and the respective antigen. PMID- 16346413 TI - Cloning and Expression of Thermostable alpha-Amylase Gene from Bacillus stearothermophilus in Bacillus stearothermophilus and Bacillus subtilis. AB - The structural gene for a thermostable alpha-amylase from Bacillus stearothermophilus was cloned in plasmids pTB90 and pTB53. It was expressed in both B. stearothermophilus and Bacillus subtilis. B. stearothermophilus carrying the recombinant plasmid produced about fivefold more alpha-amylase (20.9 U/mg of dry cells) than did the wild-type strain of B. stearothermophilus. Some properties of the alpha-amylases that were purified from the transformants of B. stearothermophilus and B. subtilis were examined. No significant differences were observed among the enzyme properties despite the difference in host cells. It was found that the alpha-amylase, with a molecular weight of 53,000, retained about 60% of its activity even after treatment at 80 degrees C for 60 min. PMID- 16346414 TI - Selection of bacteria with favorable transport properties through porous rock for the application of microbial-enhanced oil recovery. AB - This paper presents a bench-scale study on the transport in highly permeable porous rock of three bacterial species-Bacillus subtilis, Pseudomonas putida, and Clostridium acetobutylicum-potentially applicable in microbial-enhanced oil recovery processes. The transport of cells during the injection of bacterial suspension and nutrient medium was simulated by a deep bed filtration model. Deep bed filtration coefficients and the maximum capacity of cells in porous rock were measured. Low to intermediate ( approximately 10/ml) injection concentrations of cellular suspensions are recommended because plugging of inlet surface is less likely to occur. In addition to their resistance to adverse environments, spores of clostridia are strongly recommended for use in microbial-enhanced oil recovery processes since they are easiest among the species tested to push through porous rock. After injection, further transport of bacteria during incubation can occur by growth and mobility through the stagnant nutrient medium which fills the porous rock. We have developed an apparatus to study the migration of bacteria through a Berea sandstone core containing nutrient medium. PMID- 16346415 TI - Confirmation of Occurrence of Hydroxamate Siderophores in Soil by a Novel Escherichia coli Bioassay. AB - The occurrence of ferrichrome-type hydroxamate siderophores in soil was confirmed. In the presence of the iron-scavenging chelator ethylenediamine[di(o hydroxyphenylacetic)acid], soil extract stimulated the growth of an Escherichia coli strain possessing the ferrichrome transport protein (TonA) but did not stimulate growth of a strain lacking this protein (TonA). The siderophore concentration in a 1:1 (soil-water) extract was estimated to be approximately 78 nM. Specificity of the assay was supported by the absence of significant differential strain responses to ferric citrate, ferric 2,3-dihydroxybenzoate, enterochelin, ferrioxamine B, coprogen, and triacetylfusigen. PMID- 16346416 TI - Sediment nitrification, denitrification, and nitrous oxide production in a deep arctic lake. AB - We used a combination of N tracer methods and a C(2)H(2) blockage technique to determine the role of sediment nitrification and denitrification in a deep oligotrophic arctic lake. Inorganic nitrogen concentrations ranged between 40 and 600 nmol . cm, increasing with depth below the sediment-water interface. Nitrate concentrations were at least 10 times lower, and nitrate was only detectable within the top 0 to 6 cm of sediment. Eh and pH profiles showed an oxidized surface zone underlain by more reduced conditions. The lake water never became anoxic. Sediment Eh values ranged from -7 to 484 mV, decreasing with depth, whereas pH ranged from 6.0 to 7.3, usually increasing with depth. The average nitrification rate (49 ng of N . cm . day) was similar to the average denitrification rate (44 ng of N . cm . day). In situ N(2)O production from nitrification and denitrification ranged from 0 to 25 ng of N . cm . day. Denitrification appears to depend on the supply of nitrate by nitrification, such that the two processes are coupled functionally in this sediment system. However, the low rates result in only a small nitrogen loss. PMID- 16346417 TI - Binding characteristics of lectins involved in the trapping of nematodes by fungi. AB - Seventeen saccharides were tested for their ability to bind to the trap lectins of three species of nematode-trapping fungi and prevent nematode capture. The lectin of Arthrobotrys conoides was found to be inhibited by alpha-d-glucose/d mannose and similar saccharides. The lectins of Monacrosporium eudermatum and Monacrosporium rutgeriensis were inhibited by alpha-l-fucose and 2-deoxy-d glucose, respectively. Human group O(H) erythrocytes agglutinated to traps of M. eudermatum but not A. conoides or M. rutgeriensis. There was no agglutination of group A or B to traps formed by all three fungi. Exposure of the traps to trypsin eliminated the ability to capture nematodes. The presence of d-glucose/d-mannose and l-fucose residues on the nematode cuticle was suggested through the use of commercially prepared lectin-peroxidase conjugates. PMID- 16346418 TI - Denitrification and Assimilatory Nitrate Reduction in Aquaspirillum magnetotacticum. AB - Aquaspirillum magnetotacticum MS-1 grew microaerobically but not anaerobically with NO(3) or NH(4) as the sole nitrogen source. Nevertheless, cell yields varied directly with NO(3) concentration under microaerobic conditions. Products of NO(3) reduction included NH(4), N(2)O, NO, and N(2). NO(2) and NH(2)OH, each toxic to cells at 0.2 mM, were not detected as products of cells growing on NO(3). NO(3) reduction to NH(4) was completely repressed by the addition of 2 mM NH(4) to the growth medium, whereas NO(3) reduction to N(2)O or to N(2) was not. C(2)H(2) completely inhibited N(2)O reduction to N(2) by growing cells. These results indicate that A. magnetotacticum is a microaerophilic denitrifier that is versatile in its nitrogen metabolism, concomitantly reducing NO(3) by assimilatory and dissimilatory means. This bacterium appears to be the first described denitrifier with an absolute requirement for O(2). The process of NO(3) reduction appears well adapted for avoiding accumulation of several nitrogenous intermediates that are toxic to cells. PMID- 16346419 TI - Characterization of Phage-Sensitive Mutants from a Phage-Insensitive Strain of Streptococcus lactis: Evidence for a Plasmid Determinant that Prevents Phage Adsorption. AB - A phage-insensitive strain of Streptococcus lactis, designated ME2, was used as a prototype strain for the study of mechanisms and genetics of phage resistance in the lactic streptococci. Mutants sensitive to a Streptococcus cremoris phage, varphi18, were isolated at a level of 17% from cultures of ME2 after sequential transfer at 30 degrees C. Phage-sensitive mutants of ME2 were not fully permissive to varphi18. The efficiency of plating of varphi18 on the mutants was 5 x 10 as compared with <10 for varphi18 on ME2. Further characterization of the mutants showed that they efficiently adsorbed varphi18 at levels of >99.8%, whereas ME2 adsorbed only 20 to 40% of varphi18. These results suggest that increased phage susceptibility of the mutants may result from the loss of a mechanism that inhibits phage adsorption. Moreover, the high frequency of spontaneous mutation in ME2 indicates the involvement of an unstable genetic determinant in this phage defense mechanism. ME2 was shown to possess 13 plasmids ranging in size from 1.6 to 34 megadaltons. Of 40 mutants examined that had increased efficiencies of plating, all were missing a 30-megadalton plasmid, pME0030. These data suggest that pME0030 codes for a function that prevents phage adsorption. Further phenotypic characterization of the phage-sensitive mutants showed that some mutants were deficient in the ability to ferment lactose (Lac) and hydrolyze milk proteins (Prt). However, the Lac and Prt phenotype segregated independently of the phage-sensitivity phenotype. One phage-sensitive adsorption mutant, designated N1, was tested for susceptibility to 14 different phages. N1 showed increased capacity to adsorb 4 and to replicate 2 of these 14 phages, thereby indicating a phage resistance mechanism in ME2 that generalizes to phage interactions other than the specific varphi18-ME2 phage-host interaction. These data provide evidence for a unique plasmid-linked phage defense mechanism in phage-insensitive strains of lactic streptococci. PMID- 16346420 TI - Relationship Between Lignin Degradation and Production of Reduced Oxygen Species by Phanerochaete chrysosporium. AB - The relationship between the production of reduced oxygen species, hydrogen peroxide (H(2)O(2)), superoxide (O(2)), and hydroxyl radical (.OH), and the oxidation of synthetic lignin to CO(2) was studied in whole cultures of the white rot fungus Phanerochaete chrysosporium Burds. The kinetics of the synthesis of H(2)O(2) coincided with the appearance of the ligninolytic system; also, H(2)O(2) production was markedly enhanced by growth under 100% O(2), mimicking the increase in ligninolytic activity characteristic of cultures grown under elevated oxygen tension. Lignin degradation by whole cultures was inhibited by a specific H(2)O(2) scavenger, catalase, implying a role for H(2)O(2) in the degradative process. Superoxide dismutase also inhibited lignin degradation, suggesting that O(2) is also involved in the breakdown of lignin. The production of .OH was assayed in whole cultures by a benzoate decarboxylation assay. Neither the kinetics of .OH synthesis nor the final activity of its producing system obtained under 100% O(2) correlated with that of the lignin-degrading system. However, lignin degradation was inhibited by compounds which react with .OH. It is concluded that H(2)O(2), and perhaps O(2), are involved in lignin degradation; because these species are relatively unreactive per se, their role must be indirect. Conclusions about a role for .OH in ligninolysis could not be reached. PMID- 16346421 TI - Energetics of Growth of a Defined Mixed Culture of Desulfovibrio vulgaris and Methanosarcina barkeri: Interspecies Hydrogen Transfer in Batch and Continuous Cultures. AB - Interspecies hydrogen transfer was studied in Desulfovibrio vulgaris Methanosarcina barkeri mixed cultures. Experiments were performed under batch and continuous growth culture conditions. Lactate or pyruvate was used as an energy source. In batch culture and after 30 days of simultaneous incubation, these organisms were found to yield 1.5 mol of methane and 1.5 mol of carbon dioxide per mol of lactate fermented. When M. barkeri served as the hydrogen acceptor, growth yields of D. vulgaris were higher compared with those obtained on pyruvate without any electron acceptor other than protons. In continuous culture, all of the carbon derived from the oxidation of lactate was recovered as methane and carbon dioxide, provided the dilution rate was minimal. Increasing the dilution rate induced a gradual accumulation of acetate, causing acetate metabolism to cease at above mu = 0.05 h. Under these conditions all of the methane produced originated from carbon dioxide. The growth yields of D. vulgaris were measured when sulfate or M. barkeri was the electron acceptor. Two key observations resulted from the present study. First, although sulfate was substituted by M. barkeri, metabolism of D. vulgaris was only slightly modified. The coculture fermented lactate produced equimolar quantities of carbon dioxide and methane. Second, acetogenesis and methane formation from acetate were completely separable. PMID- 16346422 TI - Expression of a Lactose Transposon (Tn951) in Zymomonas mobilis. AB - The potential utility of Zymomonas mobilis as an organism for the commercial production of ethanol would be greatly enhanced by the addition of foreign genes which expand its range of fermentable substrates. We tested various plasmids and mobilizing factors for their ability to act as vectors and introduce foreign genes into Z. mobilis CP4. Plasmid pGC91.14, a derivative of RP1, was found to be transferred from Escherichia coli to Z. mobilis at a higher frequency than previously reported for any other plasmids. Both tetracycline resistance and the lactose operon from this plasmid were expressed in Z. mobilis CP4. Plasmid pGC91.14 was stably maintained in Z. mobilis at 30 degrees C but rapidly lost at 37 degrees C. PMID- 16346423 TI - Growth of a strictly anaerobic bacterium on furfural (2-furaldehyde). AB - A strictly anaerobic bacterium was isolated from a continuous fermentor culture which converted the organic constituents of sulfite evaporator condensate to methane and carbon dioxide. Furfural is one of the major components of this condensate. This furfural isolate could degrade furfural as the sole source of carbon and energy in a defined mineral-vitamin-sulfate medium. Acetic acid was the major fermentation product. This organism could also use ethanol, lactate, pyruvate, or fumarate and contained cytochrome c(3) and desulfoviridin. Except for furfural degradation, the characteristics of the furfural isolate were remarkably similar to those of the sulfate reducer Desulfovibrio gigas. The furfural isolate has been tentatively identified as Desulfovibrio sp. strain F-1. PMID- 16346424 TI - Isolation of a bacterium capable of degrading peanut hull lignin. AB - Thirty-seven bacterial strains capable of degrading peanut hull lignin were isolated by using four types of lignin preparations and hot-water-extracted peanut hulls. One of the isolates, tentatively identified as Arthrobacter sp., was capable of utilizing all four lignin preparations as well as extracted peanut hulls as a sole source of carbon. The bacterium was also capable of degrading specifically labeled [C]lignin-labeled lignocellulose and [C]cellulose-labeled lignocellulose from the cordgrass Spartina alterniflora and could also degrade [C]Kraft lignin from slash pine. After 10 days of incubation with [C]cellulose labeled lignocellulose or [C]lignin-labeled lignocellulose from S. alterniflora, the bacterium mineralized 6.5% of the polysaccharide component and 2.9% of the lignin component. PMID- 16346425 TI - Effect of Acidity on the Composition of an Indigenous Soil Population of Rhizobium trifolii Found in Nodules of Trifolium subterraneum L. AB - Acidity affected which members of an indigenous soil population of Rhizobium trifolii nodulated Trifolium subterraneum L. cv. Mt. Barker. In three experiments involving plants grown either in mineral salts agar adjusted to pH 4.8 or 6.8 and inoculated with a soil suspension or grown directly in samples of unamended soil (pH 4.8) or soil amended with CaCO(3) (pH 6.4), 121 of 151 isolates of R. trifolii were placed into four serogroups. Seventy-nine of these isolates were placed into two serogroups (6 and 36) whose nodulating ability was affected by the pH of the plant root environment. Representatives of serogroup 6 occupied the greatest percentage of the nodules at the low pH in both mineral salts agar (77%) and in unlimed soil (47 and 57%). The same serogroup was a minor nodule occupant at the higher pH in mineral salts agar (0%) and in limed soil (0 and 10%). In contrast, serogroup 36 was virtually absent in nodules formed at the low pH, whereas it was the dominant serogroup at the higher pH in both mineral salts agar (32%) and in limed soil (35 and 49%). Despite the isolates from within each serogroup being antigenically identical, separation of cellular proteins by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis revealed four and six different gel types within serogroups 6 and 36, respectively. Isolates represented by one or two gel types dominated the contribution of each serogroup to the nodule population. Further evidence for differences between isolates within each gel type were revealed from measurements of symbiotic effectiveness. PMID- 16346426 TI - Predominant bacteria in an activated sludge reactor for the degradation of cutting fluids. AB - For the first time, an activated sludge reactor, established for the degradation of cutting fluids, was examined for predominant bacteria. In addition, both total and viable numbers of bacteria in the reactor were determined so that the percentage of each predominant type in the total reactor population could be determined. Three samples were studied, and a total of 15 genera were detected. In each sample, the genus Pseudomonas and the genus Microcyclus were present in high numbers. Three other genera, Acinetobacter, Alcaligenes, and Corynebacterium, were also found in every sample but in lower numbers. In one sample, numerous appendaged bacteria were present, and one of these, the genus Seliberia, was the most predominant organism in that sample. However, in the other two samples no appendaged bacteria were detected. Six genera were found in this reactor which have not been previously reported in either cutting fluids in use or in other activated sludge systems. These genera were Aeromonas, Hyphomonas, Listeria, Microcyclus, Moraxella, and Spirosoma. None of the predominant bacteria belonged to groups of strict pathogens. PMID- 16346427 TI - Production of soy sauce koji mold spore inoculum in plastic bags. AB - An innovation is described for producing soy sauce koji mold spore inoculum by using inexpensive autoclavable plastic bags and reuseable plastic enclosures to make culture vessels. After growth, the spore mass could be dried and packaged in the same bag after removing the enclosure. Broken rice was used as the substrate for mold cultivation. Viable spore counts of 10 spores per g were obtained under optimal conditions. After drying at 50 degrees C for 6 h, the moisture content of the spore mass decreased from 35.22 to 6.32% with no significant effect on spore viability. The dry spores could be stored in the refrigerator or at room temperature for at least 3 months. PMID- 16346428 TI - Enhancement of goldfish virus type 2 replication by 1- naphthol, the major hydrolysis product of the pesticide carbaryl. AB - The major hydrolysis product of the pesticide carbaryl, 1-naphthol, enhanced the replication of goldfish virus type 2 in piscine cell culture at concentrations below any detectable cytotoxicity. Enhancement indices of 2.3, 3.7, and 7.1 were observed at 1, 5, and 10 ppm (mul/liter), respectively. Replication at 0.5 ppm was equivalent to that of controls. PMID- 16346429 TI - Influence of a calcium-specific chelant on biofilm removal. AB - This paper describes the influence of ethylene glycol-bisbeta-aminoethyl ether)- N, N-tetraacetic acid (EGTA) on biofilm removal. The addition of EGTA resulted in the immediate detachment of biofilm which suggests that the chelant removed essential calcium from the biofilm, causing it to detach. PMID- 16346430 TI - Free-living Rhizobium strain able to grow on n(2) as the sole nitrogen source. AB - [This corrects the article on p. 711 in vol. 45.]. PMID- 16346431 TI - In Situ Studies with Membrane Diffusion Chambers of Antibiotic Resistance Transfer in Escherichia coli. AB - [This corrects the article on p. 840 in vol. 44.]. PMID- 16346432 TI - Effects of environmental toxicants on metabolic activity of natural microbial communities. AB - Two methods of measuring microbial activity were used to study the effects of toxicants on natural microbial communities. The methods were compared for suitability for toxicity testing, sensitivity, and adaptability to field applications. This study included measurements of the incorporation of C-labeled acetate into microbial lipids and microbial glucosidase activity. Activities were measured per unit biomass, determined as lipid phosphate. The effects of various organic and inorganic toxicants on various natural microbial communities were studied. Both methods were useful in detecting toxicity, and their comparative sensitivities varied with the system studied. In one system, the methods showed approximately the same sensitivities in testing the effects of metals, but the acetate incorporation method was more sensitive in detecting the toxicity of organic compounds. The incorporation method was used to study the effects of a point source of pollution on the microbiota of a receiving stream. Toxic doses were found to be two orders of magnitude higher in sediments than in water taken from the same site, indicating chelation or adsorption of the toxicant by the sediment. The microbiota taken from below a point source outfall was 2 to 100 times more resistant to the toxicants tested than was that taken from above the outfall. Downstream filtrates in most cases had an inhibitory effect on the natural microbiota taken from above the pollution source. The microbial methods were compared with commonly used bioassay methods, using higher organisms, and were found to be similar in ability to detect comparative toxicities of compounds, but were less sensitive than methods which use standard media because of the influences of environmental factors. PMID- 16346433 TI - Initial phases of starvation and activity of bacteria at surfaces. AB - The activity of the hydrophilic Vibrio sp. strain DW1 and the hydrophobic Pseudomonas sp. strain S9, which both undergo starvation-induced responses, was examined at nutrient-enriched and nutrient-deficient interfaces. The initial period of response to a starvation regime ("dwarfing" phase) is a sequence of two processes: fragmentation and continuous size reduction of the fragmented cells. This dwarfing phase is also one of intense metabolic activity as supported by O(2) uptake measurements of the endogenous metabolism and the use of inhibitors of the proton flow, the electron transport chain, and membrane-bound ATPase. Hydrophilic bacteria become even smaller at nutrient-deficient surfaces than in the liquid phase upon starvation, and this is reflected in a higher endogenous metabolism exhibited by surface-associated cells compared with those in the liquid phase. On the other hand, hydrophobic bacteria dwarfing at surfaces did not exhibit a greater size reduction and exhibited an endogenous metabolism that was only slightly higher than that of cells in the liquid phase. Bacterial scavenging of surface-localized nutrients is related to the degree of irreversible binding of dwarf and starved bacteria, which in turn may be related to the degree of cell surface hydrophobicity. PMID- 16346434 TI - Initiation of Activation of a Preemergent Herbicide by a Novel Alkylsulfatase of Pseudomonas putida FLA. AB - The activation of the preemergent herbicide 2-(2,4-dichlorophenoxy)ethyl sulfate (Crag herbicide) is initiated by soil microorganisms that are presumed to act by removing the ester sulfate group via some type of sulfatase enzyme. An enrichment technique with the herbicide as the sole source of sulfur led to the isolation of several pure cultures that could produce 2-(2,4-dichlorophenoxy)ethanol from the herbicide. One of these, a strain of Pseudomonas putida, was particularly active. Polyacrylamide gel zymograms of extracts of cells grown on nutrient broth showed the presence of three secondary and three primary alkylsulfatases. One of the latter enzymes was active toward Crag herbicide as well as sodium dodecyl sulfate. Maximum activity was obtained in the late-stationary phase of growth, and enzyme yields were not affected by either the presence or the absence of the herbicide in the growth medium. The enzyme was purified 2,670-fold to homogeneity by a combination of streptomycin sulfate treatment, heat treatment, and column chromatography on DEAE-cellulose, Sephacryl 200-S, and butyl agarose. The pure enzyme was tetrameric (molecular weight, 295,000) and most active at pH 6.0. Saturation kinetics with inhibition by excess substrate were observed for Crag herbicide and octyl sulfate. 2-Butox-yethyl sulfate was a relatively poor substrate, and dodecyltriethoxy sulfate was not hydrolyzed at all. Enzymatic hydrolysis of each substrate in the presence of H(2)O led to incorporation of O exclusively into SO(4) ions in all three cases. The Crag herbicide sulfatase therefore acts by cleaving the O-S bond of the C-O-S ester linkage, in contrast with other alkylsulfatases acting on long-chain alkyl sulfates. PMID- 16346435 TI - Motility and Chemotaxis of Rhizobium meliloti in Soil. AB - The spreading of Rhizobium meliloti strains in various autoclaved soils was measured by using simple modifications of established techniques. Behavioral mutants were used to demonstrate that efficient spreading requires active motility and chemotaxis. The rate of spreading was affected by physical or chemical differences among soils and by changes in water content. PMID- 16346436 TI - Factors determining annual changes in bacterial photosynthetic pigments in holomictic lake ciso, Spain. AB - The pigments and biomass of anoxygenic phototrophic bacteria were measured during a year cycle in Lake Ciso (Girona, Spain). Two genera, Chromatium and Chlorobium, accounted for most of the bacterial population. The bacteria were present throughout the year despite complete mixing of the lake during fall and winter. This was possible because the sulfide production in the sediment was high enough to make the lake anaerobic to the very surface. Solar radiation, temperature, and biomass of Chromatium sp. were found to be important in determining pigment concentrations by correlation analysis. Sulfide concentration and biomass of Chlorobium spp. were found to be unimportant. A path analysis was performed to determine what percentage of the variability of pigments could be explained by the variables studied. Since a high percentage could be explained, it was possible to conclude that solar radiation, temperature, and biomass of Chromatium sp. were the main variables. PMID- 16346437 TI - Chemically defined medium for cultivation of several epiphytic and phytopathogenic spiroplasmas. AB - A chemically defined medium, LD82, was formulated for in vitro cultivation of spiroplasmas. Medium LD82 supported good growth for four epiphytic and insect pathogenic spiroplasmas, Spiroplasma floricola 23-6, Spiroplasma sp. strain SR3, Spiroplasma sp. strain brevi, and Spiroplasma sp. strain AS576, and of the phytopathogenic spiroplasmas Spiroplasma citri Maroc R8A2 and PC1. Titers of all six strains grown in defined medium LD82 reached 2.0 x 10 to 6.0 x 10 CFU/ml of culture. All spiroplasma strains tested formed colonies readily on agar medium LD82. None of the spiroplasmas formed typical fried-egg colonies. All formed diffuse colonies, but the forms of colonies differed somewhat among the spiroplasma strains. In preliminary studies of nutritional requirements, phospholipids slightly enhanced the growth of the epiphytic and insect-pathogenic strains in medium LD82 and were found essential for good growth of S. citri. PMID- 16346438 TI - Growth of bacteria in inorganic medium at different levels of airborne organic substances. AB - Invasion rates of airborne organic substances into sterile mineral medium were compared by using flasks closed with cotton stoppers, silicone stoppers, and screw caps with Teflon gaskets. The resulting increases of dissolved organic carbon were 0.5, 0.2, and 0 mg/liter per week, respectively. The compounds supported the growth of lake water bacteria and a strain of Pseudomonas fluorescens. Growth rates were correlated to the permeability of the stoppers used. The measured input of organic carbon in the sterile mineral medium is considered to be a minimum value for the actual contribution of organic compounds by the air. Multiplication rates of the bacteria suggest that the organisms prevent the escape of volatile organic substances from the medium by rapid utilization. The steady nutrient supply through the air should be considered in growth experiments with bacteria at low concentrations of nutrients. PMID- 16346439 TI - Corrected Version Distribution of Heterogeneous and Homologous Plasmids in Bacillus spp. AB - A total of 75 strains (including 5 reference strains) of Bacillus amyloliquefaciens, B. cereus, B. circulans, B. licheniformis, B. megaterium, B. pumilus, B. sphaericus, B. subtilis, and B. thuringiensis and 36 species unidentified Bacillus strains were surveyed for plasmids by cesium chloride ethidium bromide equilibrium centrifugation of cell lysates in a study of antibiotic resistance in host cells. Of the 111 strains, 13 (including 3 reference strains) were found to harbor plasmids, and 5 of the 13 showed antibiotic resistance. This antibiotic resistance appeared not to be due to the plasmids, however, because the trait was not cured by cultivation of cells in nutrient medium containing ethidium bromide (1 mug/ml), sodium dodecyl sulfate (0.2 mug/ml), or novobiocin (1 mug/ml), except in one strain, in which kanamycin and streptomycin resistances were cured by novobiocin. One strain of B. amyloliquefaciens, S294, was found to harbor a plasmid, pFTB14, which differed from the plasmid species of classes 1 to 6 in B. subtilis and B. amyloliquefaciens, as determined by restriction analysis and DNA contour length determination. However, in DNA-DNA hybridization on a filter after Southern blotting from an agarose gel, the pFTB14 DNA hybridized with plasmids of classes 1 to 5. Three strains of B. thuringiensis each carried at least 4 to 11 plasmid species, whereas no plasmids were detected in four strains of B. cereus, which, in relation to B. thuringiensis, is closely related taxonomically and has highly homologous DNA sequences. The plasmid DNAs prepared from species other than B. subtilis and B. amyloliquefaciens did not hybridize with that of pFTB14. PMID- 16346440 TI - Bacterial Decarboxylation of o-Phthalic Acids. AB - The decarboxylation of phthalic acids was studied with Bacillus sp. strain FO, a marine mixed culture ON-7, and Pseudomonas testosteroni. The mixed culture ON-7, when grown anaerobically on phthalate but incubated aerobically with chloramphenicol, quantitatively converted phthalic acid to benzoic acid. Substituted phthalic acids were also decarboxylated: 4,5-dihydroxyphthalic acid to protocatechuic acid; 4-hydroxyphthalic and 4-chlorophthalic acids to 3 hydroxybenzoic and 3-chlorobenzoic acids, respectively; and 3-fluorophthalic acid to 2-and 3-fluorobenzoic acids. Bacillus sp. strain FO gave similar results except that 4,5-dihydroxyphthalic acid was not metabolized, and both 3- and 4 hydroxybenzoic acids were produced from 4-hydroxyphthalic acid. P. testosteroni decarboxylated 4-hydroxyphthalate (to 3-hydroxybenzoate) and 4,5 dihydroxyphthalate but not phthalic acid and halogenated phthalates. Thus, P. testosteroni and the mixed culture ON-7 possessed 4,5-dihydroxyphthalic acid decarboxylase, previously described in P. testosteroni, that metabolized 4,5 dihydroxyphthalic acid and specifically decarboxylated 4-hydroxyphthalic acid to 3-hydroxybenzoic acid. The mixed culture ON-7 and Bacillus sp. strain FO also possessed a novel decarboxylase that metabolized phthalic acid and halogenated phthalates, but not 4,5-dihydroxyphthalate, and randomly decarboxylated 4 hydroxyphthalic acid. The decarboxylation of phthalic acid is suggested to involve an initial reduction to 1,2-dihydrophthalic acid followed by oxidative decarboxylation to benzoic acid. PMID- 16346441 TI - Aerobic and Anaerobic Catabolism of Vanillic Acid and Some Other Methoxy-Aromatic Compounds by Pseudomonas sp. Strain PN-1. AB - Vanillic acid (4-hydroxy-3-methoxybenzoic acid) supported the anaerobic (nitrate respiration) but not the aerobic growth of Pseudomonas sp. strain PN-1. Cells grown anaerobically on vanillate oxidized vanillate, p-hydroxybenzoate, and protocatechuic acid (3,4-dihydroxybenzoic acid) with O(2) or nitrate. Veratric acid (3,4-dimethoxybenzoic acid) but not isovanillic acid (3-hydroxy-4 methoxybenzoic acid) induced cells for the oxic and anoxic utilization of vanillate, and protocatechuate was detected as an intermediate of vanillate breakdown under either condition. Aerobic catabolism of protocatechuate proceeded via 4,5-meta cleavage, whereas anaerobically it was probably dehydroxylated to benzoic acid. Formaldehyde was identified as a product of aerobic demethylation, indicating a monooxygenase mechanism, but was not detected during anaerobic demethylation. The aerobic and anaerobic systems had similar but not identical substrate specificities. Both utilized m-anisic acid (3-methoxybenzoic acid) and veratrate but not o- or p-anisate and isovanillate. Syringic acid (4-hydroxy-3,5 dimethoxybenzoic acid), 3-O-methylgallic acid (3-methoxy-4,5-dihydroxybenzoic acid), and 3,5-dimethoxybenzoic acid were attacked under either condition, and formaldehyde was liberated from these substrates in the presence of O(2). The anaerobic demethylating system but not the aerobic enzyme was also active upon guaiacol (2-methoxyphenol), ferulic acid (3-[4-hydroxy-3-methoxyphenyl]-2 propenoic acid), 3,4,5-trimethoxycinnamic acid (3-[3,4,5-trimethoxyphenyl]-2 propenoic acid), and 3,4,5-trimethoxybenzoic acid. The broad specificity of the anaerobic demethylation system suggests that it probably is significant in the degradation of lignoaromatic molecules in anaerobic environments. PMID- 16346442 TI - Siderophores Produced by Nitrogen-Fixing Azotobacter vinelandii OP in Iron Limited Continuous Culture. AB - Azotobacter vinelandii requires a high complement of iron and an efficient iron acquisition system to support nitrogen fixation. To circumvent problems inherent in batch culture trace metal studies, continuous cultures were used to measure the response of A. vinelandii to iron stress. Iron was found to be growth limiting for nitrogen-fixing A. vinelandii at a concentration as high as 12.5 muM; iron was growth sufficient at 25 muM. Iron-stressed A. vinelandii in continuous culture formed 2,3-hydroxybenzoic acid (DHB), 2-N,6-N-di-(2,3 dihydroxybenzoyl)-l-lysine (DHBL), and a chromophoric yellow-green fluorescent peptide (YGFP). At a fixed dilution rate of 0.1 h, steady-state growth occurred at growth-limiting iron concentrations. DHB and DHBL were quantitatively measured during iron-limited steady states and iron-sufficient states by Arnow colorimetric assays. YGFP was determined by absorbance measurements taken at 380 nm, and the concentration was calculated from the reported specific absorption coefficient. Biomass increased and DHBL, DHB, and YGFP concentrations decreased as the concentration of growth-limiting iron was increased in the culture vessel and medium reservoirs. DHBL was the major siderophore and YGFP was the minor siderophore species produced during iron-limited equilibrium growth. A low level of DHB and YGFP, but no DHBL, was formed under iron-sufficient conditions. These results provide further physiological evidence that DHB, YGFP, and especially DHBL may function as siderophores in nitrogen-fixing A. vinelandii. PMID- 16346443 TI - Analysis of Poly-beta-Hydroxybutyrate in Rhizobium japonicum Bacteroids by Ion Exclusion High-Pressure Liquid Chromatography and UV Detection. AB - Ion-exclusion high-pressure liquid chromatography (HPLC) was used to measure poly beta-hydroxybutyrate (PHB) in Rhizobium japonicum bacteroids. The products in the acid digest of PHB-containing material were fractionated by HPLC on Aminex HPX 87H ion-exclusion resin for organic acid analysis. Crotonic acid formed from PHB during acid digestion was detected by its intense absorbance at 210 nm. The Aminex-HPLC method provides a rapid and simple chromatographic technique for routine analysis of organic acids. Results of PHB analysis by Aminex-HPLC were confirmed by gas chromatography and spectrophotometric analysis. PMID- 16346444 TI - Reproduction of Bacillus stearothermophilus as a Function of Temperature and Pressure. AB - The colony-forming ability and the rate of reproduction of Bacillus stearothermophilus were determined as a function of temperature and pressure. Colonies were formed between 39 and 70 degrees C at atmospheric pressure and between 54 and 67 degrees C at 45 MPa. Colonies did not form at 55.9 MPa. The rate of reproduction in broth cultures decreased with increasing pressure at all temperatures. The rate of reproduction diminished rapidly with pressure above 10.4 MPa. Therefore, increased hydrostatic pressure was not sufficient to enable B. stearothermophilus to function beyond the temperature limiting growth and reproduction at atmospheric pressure, and B. stearothermophilus should grow in naturally or artificially warmed regions of the deep sea, where the pressure is less than approximately 50 MPa, although growth rates would be low above 10 MPa. PMID- 16346445 TI - Distribution, population dynamics, and characteristics of ice nucleation-active bacteria in deciduous fruit tree orchards. AB - Deciduous fruit tree orchards located in the Pacific Northwest were surveyed over a 3-year period for the presence of ice nucleation-active (INA) bacteria. In the Yakima Valley, only about 30% of the fruit tree orchards contained INA bacteria (median population ca. 3 x 10 CFU/g [fresh weight]) in contrast to nearly 75% of the orchards in the Hood River Valley (median population ca. 5 x 10 CFU/g [fresh weight]). These INA populations ranged from less than 10 to over 10 CFU/g (fresh weight) of blossoms and, in Hood River Valley orchards, generally comprised over 10% of the total bacterial population. Populations of INA bacteria fluctuated during the year with highest levels developing on buds and flowers during the cool, wet spring, followed by a drop in populations during the warmer, drier, summer months and finally a gradual increase in the autumn. The INA bacteria persisted on dormant buds from which they again colonized young developing vegetative tissues. All INA bacteria were identified as Pseudomonas syringae. The frequency of ice nucleation at -5 degrees C for these strains ranged from nearly every cell being INA to less than 1 in 10 cells. The median frequency of ice nucleation at -5 degrees C was 10 cells per ice nucleus. The INA P. syringae strains from individual orchards were diverse with respect to bacteriocin typing and in ice nucleation frequency. The consistent absence of detectable INA bacteria or presence of low populations in most of the orchards surveyed during periods when critical temperatures (i.e., -2 to -5 degrees C) were common indicated a limited role for INA bacteria in frost susceptibility of most Pacific Northwest orchards. PMID- 16346446 TI - Double-staining epifluorescence technique to assess frequency of dividing cells and bacteriovory in natural populations of heterotrophic microprotozoa. AB - We have developed a double-staining procedure for use with epifluorescence microscopy which allows the detection both of dividing cells and of ingested bacteria in food vacuoles of heterotrophic microprotozoa. Microprotozoan cells are stained sequentially with the DNA-specific fluorochrome DAPI (4',6-diami-dino 2-phenylindole) and the nonspecific protein stain fluorescein isothiocyanate. During microscopic examination, heterotrophic microprotozoan cells are first located with fluorescein isothiocyanate fluorescence and then epifluorescence filter sets are switched to permit inspection under DAPI fluorescence of the cell nuclei and of the contents of food vacuoles. Among in situ populations of estuarine microprotozoa sampled over a tidal cycle, we found from 2.2 to 5.2% of the heterotrophic cells in a recognizable stage of division (nuclei elongated or double). Batch culture growth experiments were also carried out both with natural populations and with two isolated species of estuarine microprotozoa. In these experiments, the frequency of dividing cells ranged from 1.2 to 3.8% and appeared to be negatively correlated with growth rate. Microprotozoan populations sampled in continental shelf waters off Savannah, Ga., had mean frequencies of dividing cells ranging from 2.0 to 5.0%. A large fraction of cells in heterotrophic microprotozoan populations (an average of 27.4 +/- 1.0% in estuarine water and of 30.1 +/- 4.8% in shelf water) had DAPI-stained inclusions, presumably recently ingested bacteria, in their food vacuoles. PMID- 16346447 TI - Utilization of PP(i) as an Energy Source by a Clostridium sp. AB - The growth of an anaerobic, spore-forming rod we have isolated from the cockroach gut after enrichment on media containing PP(i) was stimulated by the presence of PP(i). The doubling time decreased and cell yield increased proportionately to PP(i) concentrations of up to 0.35%. A similar stimulation of the growth of Desulfotomaculum sp. by PP(i) has been reported. The PP(i)-stimulated Clostridium sp. fermented a number of sugars with the production of hydrogen, acetate, and butyrate, with smaller amounts of ethanol and butanol being produced from some substrates. The fermentation products were not qualitatively changed by the presence of PP(i), but significantly more hydrogen was produced. The organism contained several of the enzymes previously reported from Entamoeba sp. and Propionibacterium sp., in which PP(i) serves as a source of a high-energy bond in place of ATP. These include significant amounts of pyruvate-phosphate dikinase and phosphoenolpyruvate carboxytransphosphorylase. The activities of many of the catabolic enzymes of the organism, as well as of its phosphatases and pyrophosphatase, were similar whether it was grown in the presence or absence of PP(i). The organism did not accumulate intracellular polyphosphate granules but stored large amounts of glycogen. PMID- 16346448 TI - Microbial decomposition of wood in streams: distribution of microflora and factors affecting [C]lignocellulose mineralization. AB - The distribution and lignocellulolytic activity of the microbial community was determined on a large log of Douglas fir (Pseudotsuga menziesii) in a Pacific Northwest stream. Scanning electron microscopy, plate counts, and degradation of [C]lignocelluloses prepared from Douglas fir and incubated with samples of wood taken from the surface and within the log revealed that most of the microbial colonization and lignocellulose-degrading activity occurred on the surface. Labeled lignocellulose and surface wood samples were incubated in vitro with nutrient supplements to determine potential limiting factors of [C]lignocellulose degradation. Incubations carried out in a nitrogenless mineral salts and trace elements solution were no more favorable to degradation than those carried out in distilled water alone. Incubations supplemented with either (NH(4))(2)SO(4) or organic nitrogen sources showed large increases in the rates of mineralization over incubations with mineral salts and trace elements alone, with the greatest effect being observed from an addition of (NH(4))(2)SO(4). Subsequent incubations with (NH(4))(2)SO(4), KNO(3), and NH(4)NO(3) revealed that KNO(3) was the most favorable for lignin degradation, whereas all three supplements were equally favorable for cellulose degradation. Supplementation with glucose repressed both lignin and cellulose mineralization. The results reported in this study indicate that nitrogen limitation of wood decomposition may exist in streams of the Pacific Northwest. The radiotracer technique was shown to be a sensitive and useful tool for assessing relative patterns of lignocellulose decay and microbial activity in wood, along with the importance of thoroughly characterizing the experimental system before its general acceptance. PMID- 16346449 TI - Torulopsis petrophilum and Surface Activity. AB - Torulopsis petrophilum can synthesize either a glycolipid surfactant or a protein emulsifier depending on the substrate used. These compounds were not produced to facilitate the uptake of an insoluble carbon source. The glycolipids produced were identical to the mixture isolated from T. bombicola. PMID- 16346451 TI - Toxicity studies of metabolites of some fungal isolates in albino mice. AB - [This corrects the article on p. 944 in vol. 46.]. PMID- 16346450 TI - Purification and Characterization of Extracellular Proteinases of Aspergillus oryzae. AB - [This corrects the article on p. 507 in vol. 30.]. PMID- 16346452 TI - Methanogenesis from sucrose by defined immobilized consortia. AB - A bacterial consortium capable of sucrose degradation primarily to CH(4) and CO(2) was constructed, with acetate as the key methanogenic precursor. In addition, the effect of agar immobilization on the activity of the consortium was determined. The primary fermentative organism, Escherichia coli, produced acetate, formate, H(2), and CO(2) (known substrates for methanogens), as well as ethanol and lactate, compounds that are not substrates for methanogens. Oxidation of the nonmethanogenic substrates, lactate and ethanol, to acetate was mediated by the addition of Acetobacterium woodii and Desulfovibrio vulgaris. The methanogenic stage was accomplished by the addition of the acetophilic methanogen Methanosarcina barkeri and the hydrogenophilic methanogen Methanobacterium formicicum. Results of studies with low substrate concentrations (0.05 to 0.2% [wt/vol]), a growth-limiting medium, and the five-component consortium indicated efficient conversion (40%) of sucrose carbon to CH(4). Significant decreases in yields of CH(4) and rates of CH(4) production were observed if any component of the consortium was omitted. Approximately 70% of the CH(4) generated occurred via acetate. Agar-immobilized cells of the consortium exhibited yields of CH(4) and rates of CH(4) production from sucrose similar to those of nonimmobilized cells. The rate of CH(4) production decreased by 25% when cysteine was omitted from reaction conditions and by 40% when the immobilized consortium was stored for 1 week at 4 degrees C. PMID- 16346453 TI - Hydrophobicity as an adhesion mechanism of benthic cyanobacteria. AB - The capacity of benthic cyanobacteria to adhere to solid substrates was examined in terms of their cell surface properties. By using a biphasic water-hydrocarbon test system, it was demonstrated that benthic cyanobacteria from divergent habitats were all hydrophobic, whereas all the planktonic cyanobacteria tested were hydrophilic. Divalent cations were found more efficient than monovalent cations in effecting the expression of hydrophobicity. Mechanical shearing of the cell surface, as well as chemical removal of the cell wall, demonstrated that the hydrophobicity was confined to the outer surface layers. The hydrophobic sites were distributed along the whole length of the cyanobacterial filament. Hydrophilic hormogonia of benthic cyanobacteria became hydrophobic within 48 h when grown in the light; chloramphenicol, 3(3,4-dichlorophenyl)1,1 dimethylurea, or incubation in the dark prevented this transition. Hydrophobicity of Phormidium filaments was masked in late stationary phase; this effect was removed by gentle washing. PMID- 16346454 TI - Deterministic three-half-order kinetic model for microbial degradation of added carbon substrates in soil. AB - The kinetics of mineralization of carbonaceous substrates has been explained by a deterministic model which is applicable to either growth or nongrowth conditions in soil. The mixed-order nature of the model does not require a priori decisions about reaction order, discontinuity period of lag or stationary phase, or correction for endogenous mineralization rates. The integrated equation is simpler than the integrated form of the Monod equation because of the following: (i) only two, rather than four, interdependent constants have to be determined by nonlinear regression analysis, (ii) substrate or product formation can be expressed explicitly as a function of time, (iii) biomass concentration does not have to be known, and (iv) the required initial estimate for the nonlinear regression analysis can be easily obtained from a linearized form rather than from an interval estimate of a differential equation. CO(2) evolution data from soil have been fitted to the model equation. All data except those from irradiated soil gave better fits by residual sum of squares (RSS) by assuming growth in soil was linear (RSS = 0.71) as opposed to exponential (RSS = 2.87). The underlying reasons for growth (exponential versus linear), no growth, and relative degradation rates of substrates are consistent with the basic mechanisms from which the model is derived. PMID- 16346455 TI - Production of a Biosurfactant from Torulopsis bombicola. AB - Two types of carbon sources-carbohydrate and vegetable oil-are necessary to obtain large yields of biosurfactant from Torulopsis bombicola ATCC 22214. Most of the surfactant is produced in the late exponential phase of growth. It is possible to grow the yeast on a single carbon source and then add the other type of substrate, after the exponential growth phase, and cause a burst of surfactant production. This product is a mixture of glycolipids. The maximum yield is 70 g liter, or 35% of the weight of the substrate used. An economic comparison demonstrated that this biosurfactant could be produced significantly more cheaply than any of the previously reported microbial surfactants. PMID- 16346456 TI - Location of Peptidases Outside and Inside the Membrane of Streptococcus cremoris. AB - Peptidase activity determinations involving native cells of Streptococcus cremoris and completely disrupted cell preparations, as well as experiments concerned with peptidase activity distribution among cell fractions obtained by a damage-restrictive removal of the cell wall and release of intracellular material, suggest the presence of peptidases with distinguishable locations. Alanyl, leucyl, and prolyl aminopeptidase activities are most likely located in the cell wall-membrane interface, showing no detectable association with the membrane. Lysyl aminopeptidase is present not only in this location, but also as an intracellular enzyme. Endopeptidase activity and glutamate aminopeptidase activity appear to be weakly associated with the membrane. The locations of these two peptidase activities, unlike those of the former aminopeptidase activities, impose a restriction on their expression. Results of experiments concerned with permeabilization of the membrane and findings regarding an effect of the local environment of the enzymes on their pH activity profiles are evaluated and considered as being indicative of the proposed location. The possible implications of these findings with respect to protein utilization during growth of the organism in milk are discussed. PMID- 16346457 TI - Bacterial Symbionts in the Sugar Beet Root Maggot, Tetanops myopaeformis (von Roder). AB - Aerobic heterotrophic and facultative anaerobic bacteria were isolated from all developmental stages of the sugar beet root maggot, Tetanops myopaeformis (von Roder). Two distinct bacterial symbiotic relationships were observed. Serratia liquefaciens and Serratia marcescens were found to be associated with all developmental stages. Bacterial symbiont transmission occurred from one generation to the next. Symbionts were transferred from the male reproductive system to the female reproductive system, where both an internal infiltration of the egg chorion and an external smearing of the eggs occurred during oviposition. Pseudomonas maltophilia was found in association with the larval gut and the inner surface of the puparium. Electron microscopy of the inner puparial surface revealed symbionts within the chitinous wall. In vitro symbiont chitinase production was found, using both nephelometric (turbidimetric) and N acetylglucosamine assays. A relationship appeared to exist between adult fly emergence and enzymatic chitin degradation of the puparium by the bacterial symbionts. PMID- 16346458 TI - Nitrogen-Fixing Bacteria from Warty Lenticellate Bark of a Mangrove Tree, Bruguiera gymnorrhiza (L.) Lamk. AB - Detached warty lenticellate bark of a mangrove tree species, Bruguiera gymnorrhiza (L.) Lamk. from Iriomote Island, Okinawa, a subtropical region of Japan, showed development of acetylene reduction activity when incubated in a mineral nutrient solution lacking nitrogen under an atmosphere consisting of 5% O(2), 90% N(2), and 5% C(2)H(2). The bacteria responsible for nitrogen fixation were isolated from the bark, and their capacity for acetylene reduction and the incorporation of N(2) into the bacterial cells was confirmed. Four representative strains of the isolates were subjected to taxonomic classification. Two strains were similar to Enterobacter cloacae, and another resembled Enterobacter aerogenes. The characteristics of the fourth strain were similar to those of Klebsiella planticola (Bagley et al., Curr. Microbiol. 6:105-109, 1981). The results of this investigation suggest that the acetylene reduction activity of lenticellate warts of mangrove trunk bark is due to the presence in the warts of nitrogen-fixing bacteria belonging to the family Enterobacteriaceae. PMID- 16346459 TI - Microbial transformation of esters of chlorinated carboxylic acids. AB - Two groups of compounds were selected for microbial transformation studies. In the first group were carboxylic acid esters having a fixed aromatic moiety and an increasing length of the alkyl component. Ethyl esters of chlorine-substituted carboxylic acids were in the second group. Microorganisms from environmental waters and a pure culture of Pseudomonas putida U were used. The bacterial populations were monitored by plate counts, and disappearance of the parent compound was followed by gas-liquid chromatography as a function of time. The products of microbial hydrolysis were the respective carboxylic acids. Octanol water partition coefficients (K(ow)) for the compounds were measured. These values spanned three orders of magnitude, whereas microbial transformation rate constants (k(b)) varied only 50-fold. The microbial rate constants of the carboxylic acid esters with a fixed aromatic moiety increased with an increasing length of alkyl substituents. The regression coefficient for the linear relationships between log k(b) and log K(ow) was high for group 1 compounds, indicating that these parameters correlated well. The regression coefficient for the linear relationships for group 2 compounds, however, was low, indicating that these parameters correlated poorly. PMID- 16346460 TI - Mineral soils as carriers for Rhizobium inoculants. AB - Mineral soil-based inoculants of Rhizobium meliloti and Rhizobium phaseoli survived better at 4 degrees C than at higher temperatures, but ca. 15% of the cells were viable at 37 degrees C after 27 days. Soil-based inoculants of R. meliloti, R. phaseoli, Rhizobium japonicum, and a cowpea Rhizobium sp. applied to seeds of their host legumes also survived better at low temperatures, but the percent survival of such inoculants was higher than peat-based inoculants at 35 degrees C. Survival of R. phaseoli, R. japonicum, and cowpea rhizobia was not markedly improved when the cells were suspended in sugar solutions before drying them in soil. Nodulation was abundant on Phaseolus vulgaris derived from seeds that had been coated with a soil-based inoculant and stored for 165 days at 25 degrees C. The increase in yield and nitrogen content of Phaseolus angularis grown in the greenhouse was the same with soil-and peat-based inoculants. We suggest that certain mineral soils can be useful and readily available carriers for legume inoculants containing desiccation-resistant Rhizobium strains. PMID- 16346461 TI - Nodularia (cyanobacteriaceae) akinetes in the sediments of the peel-harvey estuary, Western australia: potential inoculum source for nodularia blooms. AB - The populations of viable Nodularia (Cyanobacteriaceae) propagules in the sediments of the Peel-Harvey Estuary in Western Australia were assessed over location, time, and depth. The sediments of the Harvey Estuary had greater numbers of Nodularia propagules than those of the contiguous Peel Inlet. This was consistent with the distribution of Nodularia blooms in the system. The sediment populations of Nodularia propagules has increased up to 100-fold at the study sites over a 4-year period during which three blooms have occurred. It is considered that the majority of the propagules are akinetes. The significance of the sediment akinetes in providing the inoculum for rapid onset of Nodularia blooms is discussed. The population of akinetes decreased with depth, but viable akinetes were still found at 35 cm, the maximum depth sampled. Bioturbation by polychaete worms is likely to be significant in the distribution of akinetes to these depths. PMID- 16346462 TI - Application of a microcomputer-based system to control and monitor bacterial growth. AB - A modular microcomputer-based system was developed to control and monitor various modes of bacterial growth. The control system was composed of an Apple II Plus microcomputer with 64-kilobyte random-access memory; a Cyborg ISAAC model 91A multichannel analog-to-digital and digital-to-analog converter; paired MRR-1 pH, pO(2), and foam control units; and in-house-designed relay, servo control, and turbidimetry systems. To demonstrate the flexibility of the system, we grew bacteria under various computer-controlled and monitored modes of growth, including batch, turbidostat, and chemostat systems. The Apple-ISAAC system was programmed in Labsoft BASIC (extended Applesoft) with an average control program using ca. 6 to 8 kilobytes of memory and up to 30 kilobytes for datum arrays. This modular microcomputer-based control system was easily coupled to laboratory scale fermentors for a variety of fermentations. PMID- 16346463 TI - In Vivo Cloning of lac Genes in Streptococcus lactis ML3. AB - The isolation and characterization of a Streptococcus lactis ML3 strain which possessed a recombinant lactose plasmid is described. The recombination events generating this plasmid occurred in vivo in a recombination-deficient strain and appeared to be mediated by transposition events. Restriction mapping revealed that the recombinant plasmid, pDA0307, contained a region of the lactose plasmid, pSK08, linked to another resident plasmid, pSK07. Copy number determinations indicated that the lac genes were present at approximately 20 copies per cell in pDA0307, whereas the lac genes are normally present at approximately 10 copies per cell in pSK08. The strain containing pDA0307 displayed a 21 to 54% increase in the expression of the Lac enzyme phospho-beta-d-galactosidase. However, the strain containing pDA0307 both grew and produced lactic acid in milk at rates identical to that of a strain containing pSK08. This result suggests that lac gene dosage of plasmid-linked lac genes was not limiting the rate at which these derivatives of S. lactis ML3 fermented milk. PMID- 16346464 TI - Genetic Construction of Lactose-Utilizing Xanthomonas campestris. AB - Xanthomonas campestris, the producer of xanthan gum, possesses a beta galactosidase of very low specific activity. Plasmid pGC9114 (RP1::Tn951), generated by the transposition of the lactose transposon Tn951 to RP1, was conjugally transferred into XN1, a nalidixic acid-resistant derivative of X. campestris NRRL B-1459S-4L. Transfer occurred on membrane filters and in broth. The beta-galactosidase gene of Tn951 was expressed in X. campestris. The specific activity of beta-galactosidase in transconjugants was over 200-fold higher than that in XN1, and transconjugants grew as well in lactose-based media as in glucose-based media. The lactose-utilizing transconjugants could potentially be used to produce xanthan gum from cheese whey. PMID- 16346465 TI - Effects of Nitrapyrin [2-Chloro-6-(Trichloromethyl) Pyridine] on the Obligate Methanotroph Methylosinus trichosporium OB3b. AB - Nitrapyrin inhibited growth, CH(4) oxidation, and NH(4) oxidation, but not the oxidation of CH(3)OH, HCHO, or HCOONa, by Methylosinus trichosporium OB3b, suggesting that nitrapyrin acts against the methane monooxygenase enzyme system. The inhibition of CH(4) oxidation could be reversed by repeated washing of nitrapyrin-inhibited cells, indicating that its effect is bacteriostatic. The addition of Cu did not release the inhibition. Methane oxidation was also inhibited by 6-chloro-2-picoline. These data suggest that the mode of action of nitrapyrin on M. trichosporium is different from that on chemoautotrophic NH(4) oxidizers or methanogens. PMID- 16346466 TI - Fermentation of 6-Deoxyhexoses by Bacillus macerans. AB - Under anaerobic conditions Bacillus macerans ATCC 7068 fermented 6-deoxyhexoses (l-rhamnose, l-fucose, and d-fucose) to a mixture of 1,2-propanediol (PD), acetone, H(2), CO(2), and ethanol. The final PD concentration was proportional to the amount of l-rhamnose fermented ( approximately 0.9 mol of PD per mol of rhamnose). PD was not produced from hexoses (e.g., d-glucose or l-mannose), despite active fermentation of these substrates. Relative to the fermentation of d-glucose, the fermentation of l-rhamnose was accompanied by a twofold reduction in yield of H(2), CO(2), and cell mass. Exposure of cell extracts to l-rhamnose resulted in the transient appearance of an aldehyde intermediate. Cell extracts contained a pyridine nucleotide-linked lactaldehyde reductase activity which converted synthetic d- or l-lactaldehyde to PD. The data suggest an Embden Meyerhof pathway for 6-deoxyhexose catabolism, with the formation of lactaldehyde by a conventional aldolase cleavage reaction and subsequent reduction to PD. PMID- 16346467 TI - Aerobic microbial degradation of glucoisosaccharinic Acid. AB - alpha-Glucoisosaccharinic acid (GISA), a major by-product of kraft paper manufacture, was synthesized from lactose and used as the carbon source for microbial media. Ten strains of aerobic bacteria capable of growth on GISA were isolated from kraft pulp mill environments. The highest growth yields were obtained with Ancylobacter spp. at pH 7.2 to 9.5. GISA was completely degraded by cultures of an Ancylobacter isolate. Ancylobacter cell suspensions consumed oxygen and produced carbon dioxide in response to GISA addition. A total of 22 laboratory strains of bacteria were tested, and none was capable of growth on GISA. GISA-degrading isolates were not found in forest soils. PMID- 16346468 TI - Phosphate Nutrition of Rhizobium spp. AB - This study was conducted to determine the behavior of 40 strains from six species of Rhizobium in liquid defined media containing orthophosphate at levels likely to be encountered naturally, ranging from the high concentrations expected in nodules and artificial media to the low concentrations of soil solutions. Storage capacity in strains with high levels (2 mM) of P and ability to utilize this stored P for growth after transfer to low levels (0.06 muM) of P varied with each strain. Storage varied from about 1 to 2% P (dry weight) for all strains, with the number of generations supported dependent on the quantity of P stored and on the utilization efficiency. The ability to store P at high levels is probably less important than the uptake and utilization efficiency of P supplied at low levels. Strains varied greatly in tolerance to low levels of P maintained in solution by an iron oxide buffering system. Differences in growth rate at low levels of P were large enough to be agronomically important. PMID- 16346469 TI - Cloning the Gene for the Malolactic Fermentation of Wine from Lactobacillus delbrueckii in Escherichia coli and Yeasts. AB - The gene responsible for the malolactic fermentation of wine was cloned from the bacterium Lactobacillus delbrueckii into Escherichia coli and the yeast Saccharomyces cerevisiae. This gene codes for the malolactic enzyme which catalyzes the conversion of l-malate to l-lactate. A genetically engineered yeast strain with this enzymatic capability would be of considerable value to winemakers. L. delbrueckii DNA was cloned in E. coli on the plasmid pBR322, and two E. coll clones able to convert l-malate to l-lactate were selected. Both clones contained the same 5-kilobase segment of L. delbrueckii DNA. The DNA segment was transferred to E. coli-yeast shuttle vectors, and gene expression was analyzed in both hosts by using enzymatic assays for l-lactate and l-malate. When grown nonaerobically for 5 days, E. coli cells harboring the malolactic gene converted about 10% of the l-malate in the medium to l-lactate. The best expression in S. cerevisiae was attained by transfer of the gene to a shuttle vector containing both a yeast 2-mum plasmid and yeast chromosomal origin of DNA replication. When yeast cells harboring this plasmid were grown nonaerobically for 5 days, ca. 1.0% of the l-malate present in the medium was converted to l lactate. The L. delbrueckii controls grown under these same conditions converted about 25%. A laboratory yeast strain containing the cloned malolactic gene was used to make wine in a trial fermentation, and about 1.5% of the l-malate in the grape must was converted to l-lactate. Increased expression of the malolactic gene in wine yeast will be required for its use in winemaking. This will require an increased understanding of the factors governing the expression of this gene in yeasts. PMID- 16346470 TI - Elucidation of Growth Inhibition and Acetic Acid Production by Clostridium thermoaceticum. AB - The production of acetic acid by Clostridium thermoaceticum was studied by using batch fermentations. In a pH-controlled fermentation with sodium hydroxide (pH 6.9), this organism was able to produce 56 g of acetic acid per liter. On the other hand, when the pH was not controlled and was decreased during fermentation to 5.4, the maximum attainable acetic acid concentration was only 15.3 g/liter. To obtain a better understanding of the end product inhibition, various salts were tested to determine their effect on the growth rate of C. thermoaceticum. An inverse linear relationship between the growth rate and the final cell concentration to the sodium acetate concentration was found. By using different concentrations of externally added sodium salts, the relative growth inhibition caused by the anion was found to be in the order of acetate > chloride > sulfate. Various externally added cations of acetate were also examined with respect to their inhibitory effects on growth. The relative magnitude of inhibition on the growth rate was found to be ammonium > potassium > sodium. The combined results have shown that the undissociated acetic acid was much more inhibitory than the ionized acetate ion. Complete growth inhibition resulted when the undissociated acetic acid concentration was between 0.04 and 0.05 M and when the ionized acetate concentration was 0.8 M. Therefore, at low pH (below 6.0), undissociated acetic acid is responsible for growth inhibition, and at high pH (above 6.0), ionized acetate ion is responsible for growth inhibition. PMID- 16346471 TI - Conidium germination rate in wild and domesticated yellow-green aspergilli. AB - Conidia of domesticated yellow-green aspergilli from strains of Aspergillus oryzae (Ahlburg) Cohn and Aspergillus sojae Sakaguchi and Yamada ex Murakami, used in the preparation of koji inoculum, germinate approximately 3 h sooner than conidia produced by related wild species, Aspergillus flavus Link ex Fr. and Aspergillus parasiticus Speare. There was no consistent relationship between average conidium size and estimated 50% germination time. Germination trials were conducted on Czapek agar at 28 degrees C. A hypothesis is offered that, in the propagation of koji inoculum, selection has favored those individuals capable of rapid conidium germination and germ tube extension, attributes that enable them to gain the available substrate during intraspecific competition. PMID- 16346472 TI - Salt stress control of intracellular solutes in streptomycetes indigenous to saline soils. AB - Actinomycetes were isolated from a number of saline and saline-sodic California soils. From these isolates, two species of Streptomyces (S. griseus and S. californicus) were selected to assess their physiological response to salinity. NaCl was more inhibitory to growth rates and specific growth yields than were equivalent concentrations of KCl. Intracellular concentrations of the free amino acid pool increased in response to salt stress. Whereas the neutral free amino acids proline, glutamine, and alanine accumulated as salinity increased, concentrations of the acidic free amino acids glutamate and aspartate were reduced. Accumulation of free amino acids by streptomycetes under salt stress suggests a response typical of procaryotes, although the specific amino acids involved differ from those associated with other gram-positive bacteria. Above a salinity threshold of about 0.75 M (-3.8 MPa), there was little further intracellular accumulation of free amino acids, whereas accumulation of K salts sharply increased. PMID- 16346473 TI - Persistence of Nosema locustae Spores in Soil as Determined by Fluorescence Microscopy. AB - Nosema locustae, a protozoan parasite of grasshoppers, is used as a bioinsecticide. In the present study, the persistence of N. locustae spores in soil and the interaction of these spores with the indigenous soil microflora were examined with various forms of microscopy and staining. Fluorescence microscopy was found to be better than phase-contrast or bright-field microscopy for detecting and viewing spores in soil. Fluorescein isothiocyanate was a better fluorescent stain than acridine orange or fluorescein diacetate; water-soluble aniline blue did not stain spores. The eight bright-field microscopy stains tested (phenolic erythrosin, phenolic rose bengal, malachite green, crystal violet, safranin, Congo red, methyl red, and eosin B) were not satisfactory, as spore staining characteristics were either poor or masked by overstained soil debris. A procedure was developed which allowed spores to be extracted from soil with a peptone-phosphate buffer, recovered on a membrane filter, and stained with fluorescein isothiocyanate for microscopic counting. This procedure was used to assess the persistence of N. locustae spores in field and laboratory soils. The number of N. locustae spores in a laboratory model soil system persisted at a high level for over 8 weeks when the soil was incubated at 5 degrees C but exhibited a 1,000-fold decrease after 1 week of incubation at 27 degrees C. Persistence was related to the temperature-dependent activity of the indigenous soil microflora, which, on the basis of microscopic observations, appeared to prey on N. locustae spores. N. locustae spores were detected in an N. locustae treated field soil at a low level consistent with the level for laboratory soil incubated at 27 degrees C, and they persisted at this level for over 2 months. No spores were detected on vegetation from this field or in the soil from an adjacent, nontreated control field. N. locustae-like spores were also detected in soil from nontreated fields supporting large grasshopper populations. PMID- 16346474 TI - Competition for sulfate and ethanol among desulfobacter, desulfobulbus, and desulfovibrio species isolated from intertidal sediments. AB - Competition for sulfate and ethanol among Desulfobacter, Desulfobulbus, and Desulfovibrio species isolated from estuarine sediments was studied in energy limited chemostats. Desulfovibrio baculatus was the most successful competitor for limiting amounts of sulfate and ethanol, followed by Desulfobulbus propionicus. The success of Desulfovibrio baculatus was dependent on the availability of sufficient iron. Of the three species studied, Desulfobacter postgatei was the least successful competitor for limiting amounts of sulfate. Although stimulating the growth of Desulfobacter postgatei, addition of Ca saturated illite particles to culture media did not affect the outcome of competition for sulfate. Thus, under sulfate limitation acetate accumulated. This phenomenon was briefly discussed in relation to the flow of electrons during anaerobic mineralization in marine and estuarine sulfate-limited sediments. PMID- 16346475 TI - Transient Responses of Glucose-Limited Cultures of Cytophaga johnsonae to Nutrient Excess and Starvation. AB - Cells from glucose-limited chemostat cultures of Cytophaga johnsonae were subjected to a sudden relaxation of substrate limitation by injecting the cells into fresh batch cultures. Starvation experiments were carried out by injecting glucose-limited cells into batch cultures lacking glucose. Transient responses of biomass, glucose uptake and mineralization, ATP content, and viability on different agar media were monitored during these nutrient-shift experiments. Cells reacted differently depending on growth rate and time spent in the chemostat. Fast-growing cells showed an immediate adaptation to the new growth conditions, despite some initial overshoot reactions in ATP and uptake potential. In contrast, slowly growing cells and long-term-adapted cells showed extensive transient growth responses. Glucose uptake and mineralization potentials changed considerably during the transient growth phase before reaching new levels. During the starvation experiments, all cell types displayed a fast decrease in ATP, but the responses of the substrate uptake and mineralization potentials were strongly dependent upon the previous growth rate. Both potentials decreased rapidly in cells with high growth rates. On the other hand, cells with low growth rates maintained 80% of their uptake and mineralization potentials after 8 h of starvation. Thus, slowly growing cells are much better adapted for starvation than are fast-growing cells. PMID- 16346476 TI - Protoplast Fusion of Trichoderma reesei, Using Immature Conidia. AB - Protoplast fusion of strains derived from Trichoderma reesei QM9414 and QM9136 and the segregation of the resulting fusants were studied. Combinations of protoplasts prepared from young conidia with double amino acid requirements, one of which was a common requirement and the other uncommon, were fused in the presence of polyethylene glycol 6000. Fusants were selected as regenerant colonies requiring only the commonly deficient amino acid. The frequency of fusion was 0.9 x 10 to 4.0 x 10 for the starting conidia and 3.0 x 10 to 4.9 x 10 for the regenerated protoplasts, which was significantly higher than the expected reversion frequencies by mutation. Conidia generated on the fusant colonies showed diverse phenotypes, i.e., parental types (40 to 80%) and nonparental types (20 to 60%). Colonies developed from single conidia of the nonparental phenotype contained special spots called "knobs" that have a higher density of mycelia. The phenotype of the knobs was again varied among prototrophs, parental types, and recombinant types; and their traits were inherited stably. The phenotype of the mycelia in the nonknob part was essentially the same as that of the original conidia and again formed knobs in colonies upon transfer of a piece of mycelia to a fresh medium. The conidial DNA content of the knob clone was almost the same as that of the parents, but that of the fusants was 1.2 to 2.0 times higher than that of the parents. From these results, we conclude that knobs are the segregants from the fusants. One knob clone showed twice the carboxymethyl cellulose hydrolyzing activity of the parents, suggesting the possibility of breeding T. reesei cells by the protoplast fusion technique. PMID- 16346477 TI - Preparation, characterization, and microbial degradation of specifically radiolabeled [C]lignocelluloses from marine and freshwater macrophytes. AB - Specifically radiolabeled [C-lignin]lignocelluloses were prepared from the aquatic macrophytes Spartina alterniflora, Juncus roemerianus, Rhizophora mangle, and Carex walteriana by using [C]phenylalanine, [C]tyrosine, and [C]cinnamic acid as precursors. Specifically radiolabeled [C-polysaccharide]lignocelluloses were prepared by using [C]glucose as precursor. The rates of microbial degradation varied among [C-lignin]lignocelluloses labeled with different lignin precursors within the same plant species. To determine the causes of these differential rates, [C-lignin]lignocelluloses were thoroughly characterized for the distribution of radioactivity in nonlignin contaminants and within the lignin macromolecule. In herbaceous plants, significant amounts (8 to 24%) of radioactivity from [C]phenylalanine and [C]tyrosine were found associated with protein, although very little (3%) radioactivity from [C]cinnamic acid was associated with protein. Microbial degradation of radiolabeled protein resulted in overestimation of lignin degradation rates in lignocelluloses derived from herbaceous aquatic plants. Other differences in degradation rates among [C lignin]lignocelluloses from the same plant species were attributable to differences in the amount of label being associated with ester-linked subunits of peripheral lignin. After acid hydrolysis of [C-polysaccharide]lignocelluloses, radioactivity was detected in several sugars, although most of the radioactivity was distributed between glucose and xylose. After 576 h of incubation with salt marsh sediments, 38% of the polysaccharide component and between 6 and 16% of the lignin component (depending on the precursor) of J. roemerianus lignocellulose was mineralized to CO(2); during the same incubation period, 30% of the polysaccharide component and between 12 and 18% of the lignin component of S. alterniflora lignocellulose was mineralized. PMID- 16346479 TI - Mutation and Selection of Lactobacillus plantarum Strains That Do Not Produce Carbon Dioxide from Malate. AB - A differential medium was developed to distinguish between malate-decarboxylating (MDC) and -non-decarboxylating (MDC) strains of Lactobacillus plantarum. MDC strains produced a visible acid reaction in the medium, whereas MDC strains did not. Use of the medium allowed for rapid screening and isolation of mutagenized cells that had lost the ability to produce CO(2) from malate. PMID- 16346478 TI - Kinetics of Sulfate and Acetate Uptake by Desulfobacter postgatei. AB - The kinetics of sulfate and acetate uptake was studied in the sulfate-reducing bacterium Desulfobacter postgatei (DSM 2034). Kinetic parameters (K(m) and V(max)) were estimated from substrate consumption curves by resting cell suspensions with [S]sulfate and [C]acetate. Both sulfate and acetate consumption followed Michaelis-Menten saturation kinetics. The half-saturation constant (K(m)) for acetate uptake was 70 muM with cells from either long-term sulfate- or long-term acetate-limited chemostat cultures. The average K(m) value for sulfate uptake by D. postgatei was about 200 muM. K(m) values for sulfate uptake did not differ significantly when determined with cells derived either from batch cultures or sulfate- or acetate-limited chemostat cultures. Acetate consumption was observed at acetate concentrations of C(2) > C(3) > C(4). The carbazole-enriched culture was still able to degrade n-alkanes, isoprenoids, aromatic hydrocarbons, and sulfur heterocycles in the crude soil. PMID- 16346525 TI - Autotrophic Growth of Gas Vacuolate Strains of Microcyclus aquaticus on Methanol and Hydrogen. AB - Seven strains of Microcyclus aquaticus were found to be capable of growth on methanol and hydrogen as energy sources. Carbon was incorporated as CO(2) via the Calvin cycle, as shown by the presence of ribulosebisphosphate carboxylase activity in methanol-grown cells and by the absence of key enzymes of the ribulose monophosphate and serine pathways. In addition, incoporation of [C]methanol into cells was diminished when cultures were incubated in gas atmospheres enriched with carbon dioxide. PMID- 16346526 TI - Modification of the gelatin-matrix method for enumeration of respiring bacterial cells for use with salt-marsh water samples. AB - In this report I describe a modification of the gelatin-matrix method for determining the percentages of bacterial cells having active electron transport systems. This modification is one-step fluorochroming followed by use of an antioxidant solution instead of the secondary gelatin layer. The modified technique is more successfully applicable to water samples containing high densities of flocculant particulate matter, owing to the brighter green fluorescence of the cells, which contrasts with the red-orange fluorescence of the floc. PMID- 16346527 TI - Effect of Plasmid pIJ1008 from Rhizobium leguminosarum on Symbiotic Function of Rhizobium meliloti. AB - Plasmid pIJ1008, which carries determinants for uptake hydrogenase (Hup) activity, was transferred from Rhizobium leguminosarum to Rhizobium meliloti without impairing the capacity of the latter species to form root nodules on alfalfa. The plasmid was still present in rhizobia reisolated from the root nodules of 12 different alfalfa cultivars, but only low levels of Hup activity were detected in alfalfa. PMID- 16346528 TI - Composition of Regenerated Cell Walls of Reverting Aureobasidium pullulans Protoplasts. AB - Protoplasts of Aureobasidium pullulans, formed by treating normal blastospores with polysaccharide-hydrolyzing enzymes, synthesized glycans when incubated in shallow culture. The polysaccharides formed a loose, thick coating on the cells. The glycans that were formed are similar to the glycans in normal cell walls; however, they lack the branching that is normally found. This result is consistent with the findings of others who have studied the cell wall composition of reverting yeast protoplasts. PMID- 16346529 TI - Quantitative method for colorimetric determination of formate in fermentation media. AB - The effects of fermentation products and media supplements on a colorimetric assay for formate were evaluated. Formate was detected at concentrations as low as 0.5 mM in fermentation media. PMID- 16346530 TI - Antimicrobial effects of ionizing radiation on artificially and naturally contaminated cacao beans. AB - With an initial microbial level of ca. 10 microorganisms per g of Ivory Coast cacao beans, 5 kGy of gamma radiation under an atmosphere of air reduced the microflora per g by 2.49 and 3.03 logs at temperatures of 35 and 50 degrees C, respectively. Bahia cacao beans were artificially contaminated with dried spores of Aspergillus flavus and Penicillium citrinum, giving initial fungal levels of 1.9 x 10 and 1.4 x 10 spores per g of whole Bahia cacao beans, respectively. The average D(10) values for A. flavus and P. citrinum spores on Bahia cacao beans were 0.66 and 0.88 kGy, respectively. PMID- 16346531 TI - Hypochlorite Injury of Clostridium botulinum Spores Alters Germination Responses. AB - [This corrects the article on p. 1361 in vol. 45.]. PMID- 16346532 TI - Observations on the distinction between oligotrophic and eutrophic marine bacteria. AB - The nutritional requirements of two marine bacteria designated as oligotrophic because they could grow on media containing 10 mg of C per liter supplied as peptone and two classified as eutrophic because they could grow only at higher concentrations of C supplied as peptone were examined. Each of the four organisms was found to have its own unique group of compounds which could serve either individually or in combination as sources of carbon and energy for growth. When the peptone in the medium was replaced by another appropriate source of carbon and energy, the difference in the capacity of the organisms examined to grow at 10 mg of C per liter disappeared, and all four organisms could be described as being oligotrophic. Some of the organisms required a low concentration of one specific carbon source but a higher concentration of another. One of the organisms was inhibited by high concentrations of one specific carbon source but not by another. The observations indicate that current methods of enumeration based on the capacity of cells to grow in the presence of high or low concentrations of complex mixtures of nutrients such as peptone do not distinguish between two broad classes of bacteria differing intrinsically in their ability to grow at high and low concentrations of nutrients. Whether two such broad classes exist seems extremely doubtful. Which organisms will multiply in a particular environment will depend on both the specific nutrients available and their concentrations. PMID- 16346533 TI - Liquid chromatographic-fluorescence determination of ammonia from nitrogenase reactions: a 2-min assay. AB - The analytical potential of the reaction of ammonia with o-phthalaldehyde mercaptoethanol reagent at pH 7 (an atypical fluorescence) has already been demonstrated. This, coupled with additional findings reported here, has led to an ammonia determination well suited to nitrogenase studies. As a result, large numbers of samples can be rapidly analyzed by high-pressure liquid chromatrography methods under mild conditions and without prior microdiffusion. Neither sodium dithionite (or other components of the usual nitrogenase assay), nor alternative substrates (cyanide, azide, methyl isonitrile), nor their products (methylamine, dimethylamine, hydrazine) interfere. High-pressure liquid chromatography showed that the fluorescent "product" of the o-phthalaldehyde mercaptoethanol reagent-ammonia reaction was, in fact, more than just a single compound. Despite this, once the proper solvent composition was found, high pressure liquid chromatography with a small inexpensive C(18) "guard" column proved quite fast and reproducible for this measurement. Fluorescence response to ammonia was linear to at least 40 nmol/ml. A previous problem, long-term stability of the fluorescence, was solved by running the reactions in the dark. Background ammonia in the buffer could be substantially reduced by an analogous o phthalaldehyde mercaptoethanol reagent reaction, using t-butyl mercaptan, and solvent extraction. PMID- 16346534 TI - DNA-DNA Homology Between Lactic Streptococci and Their Temperate and Lytic Phages. AB - Temperate phages were induced from Streptococcus cremoris R(1), BK(5), and 134. DNA from the three induced phages was shown to be homologous with prophage DNA in the bacterial chromosomes of their lysogenic hosts by the Southern blot hybridization technique. P-labeled DNA from 11 lytic phages which had been isolated on cheese starters was similarly hybridized with DNA from 36 strains of lactic streptococci. No significant homology was detected between the phage and bacterial DNA. Phages and lactic streptococci used included phages isolated in a recently opened cheese plant and all the starter strains used in the plant since it commenced operation. The three temperate phages were compared by DNA-DNA hybridizations with 25 lytic phages isolated on cheese starters. Little or no homology was found between DNA from the temperate and lytic phages. In contrast, temperate phages showed a partial relationship with one another. Temperate phage DNA also showed partial homology with DNA from a number of strains of lactic streptococci, many of which have been shown to be lysogenic. This suggests that many temperate phages in lactic streptococci may be related to one another and therefore may be homoimmune with one another. These findings indicate that the release of temperate phages from starter cells currently in use is unlikely to be the predominant source of lytic phages in cheese plants. PMID- 16346535 TI - Response of an antarctic lake heterotrophic community to high dissolved oxygen. AB - The upper waters of Lake Hoare, Antarctica, contain dissolved oxygen at about three times the normal saturation (>/=42 mg liter). The response of the heterotrophic plankton community to this high dissolved oxygen was evaluated by the criteria of CFU and d-[U-C]glucose assimilated-respired. High dissolved oxygen was not inhibitory to d-[U-C]glucose assimilation-respiration compared with normal atmospheric dissolved oxygen in Lake Hoare water. The d-[U-C]glucose was assimilated and respired optimally at 12 degrees C in Lake Hoare. The d-[U C]glucose assimilated-respired in the upper saturated atmospheric dissolved oxygen waters of Mountain Lake, Va., was inhibited in contrast to Lake Hoare (P < 0.05). CFU formation was inhibited in both lakes. CFU represent <1% of the fluorochrome-stained direct counts in Lake Hoare. Lake Hoare planktobacteria are smaller than the planktobacteria in Mountain Lake. ATP size fractionation revealed that 39% of the ATP biomass was <0.5 mum in Lake Hoare. PMID- 16346536 TI - Dependence of betaine stimulation of vitamin b(12) overproduction on protein synthesis. AB - The betaine-stimulated differential synthesis of vitamin B(12), i.e., the increase in B(12) per increase in dry cell weight, by Pseudomonas denitrificans was inhibited by rifampin and chloramphenicol but not by benzylpenicillin and carbenicillin at concentrations of antibiotic that inhibit growth. The level of the first enzyme of corrin (and porphyrin) biosynthesis, delta-aminolevulinic acid synthetase, was decreased to a much greater degree by rifampin and chloramphenicol than by the penicillins. These data support the concept that betaine stimulation of B(12) synthesis is a result of its stimulation of synthesis of delta-aminolevulinic acid synthetase, a labile and presumably rate limiting enzyme of corrin formation requiring continuous induction. In further support of this hypothesis, it was found that chloramphenicol immediately interfered with both vitamin B(12) and delta-aminolevulinic acid synthetase formation, no matter when it was added to the system. PMID- 16346537 TI - Interaction of Rhizobium sp. with Toxin-Producing Fungus in Culture Medium and in a Tropical Soil. AB - Experiments were conducted to determine the influence of a toxin-producing fungus on a rhizobial population in yeast-mannitol medium and in a tropical soil. The fungus, which was isolated from a highly weathered soil (Tropeptic Eutrustox), was identified as a Metarhizum sp. The density of rhizobial populations established in yeast-mannitol medium in the absence of the fungus was 10 times higher than that established in its presence. However, the fungus did not exert similar antagonistic influence on the rhizobial population incubated with it in the sterilized test soil. Rhizobial growth activity in yeast-mannitol medium was also insensitive to the presence of the fungus when the medium was amended with 1% (wt/vol) kaolinite or montmorillonite. The results suggest that clay minerals may be responsible for protecting rhizobia against toxin-producing fungi in soil. PMID- 16346538 TI - Chemical Characterization of the Lipopolysaccharide of Pseudomonas solanacearum. AB - The carbohydrates present in lipopolysaccharide (LPS) from Pseudomonas solanacearum are rhamnose, xylose, 2-amino-2-deoxyglucose, glucose, heptose, and 2-keto-3-deoxyoctonate. LPS extracted from cultures grown on either glycerol or glucose (as the major source of carbon) and extracted after various incubation periods had similar compositions. The LPS from several strains of the bacterium contained the same component sugars, but the amounts of each sugar varied considerably. It was observed, however, that xylose and 2-amino-2-deoxyglucose increased proportionately with rhamnose, the major component. Phenol-water extracted LPS contained measurable amounts of nucleic acid, protein, and arabinan, but none of these polymers were detected in LPS extracted with phenol chloroform-petroleum ether. Polysaccharides liberated from LPS by mild acid hydrolysis were purified by gel filtration. Carbohydrate analysis of the LPS from a virulent, fluidal strain (K60) showed that the O-specific antigen consisted of rhamnose, xylose, and 2-amino-2-deoxyglucose in the proportions 4:1:1. The LPS of an avirulent, afluidal strain (B1) lacked the O-specific antigen; the R-core region consisted of rhamnose, glucose, heptose, and 2-keto-3-deoxyoctonate. Methylation analysis indicated that the K60 O-specific antigen was composed of a hexasaccharide repeating unit containing 3-, 2-, and 3,4-substituted rhamnopyranosyl residues, 3-substituted 2-amino-2-deoxyglucose, and terminal xylopyranose in the molar ratios 2:1:1:1:1. PMID- 16346539 TI - Denitrification in san francisco bay intertidal sediments. AB - The acetylene block technique was employed to study denitrification in intertidal estuarine sediments. Addition of nitrate to sediment slurries stimulated denitrification. During the dry season, sediment-slurry denitrification rates displayed Michaelis-Menten kinetics, and ambient NO(3) + NO(2) concentrations ( NO(2) were calculated to be 50 +/- 9 kJ mol for Brookston clay and 55 +/- 13 kJ mol for Fox sandy loam. The overall reaction, NO(2) --> NO (chemodenitrification), in both sterile soils was accurately first order with respect to NO(2); the activation energy was 70 +/- 2.8 kJ mol in Brookston clay and 79 +/- 1.2 kJ mol in the sandy loam, and the preexponential factors were (2.3 +/- 1.2) x 10 and (5.7 +/- 1.2) x 10 min, respectively. PMID- 16346552 TI - Methanosarcina acetivorans sp. nov., an Acetotrophic Methane-Producing Bacterium Isolated from Marine Sediments. AB - A new acetotrophic marine methane-producing bacterium that was isolated from the methane-evolving sediments of a marine canyon is described. Exponential phase cultures grown with sodium acetate contained irregularly shaped cocci that aggregated in the early stationary phase and finally differentiated into communal cysts that released individual cocci when ruptured or transferred to fresh medium. The irregularly shaped cocci (1.9 +/- 0.2 mm in diameter) were gram negative and occurred singly or in pairs. Cells were nonmotile, but possessed a single fimbria-like structure. Micrographs of thin sections showed a monolayered cell wall approximately 10 nm thick that consisted of protein subunits. The cells in aggregates were separated by visible septation. The communal cysts contained several single cocci encased in a common envelope. An amorphous form of the communal cyst that had incomplete septation and internal membrane-like vesicles was also present in late exponential phase cultures. Sodium acetate, methanol, methylamine, dimethylamine, and trimethylamine were substrates for growth and methanogenesis; H(2)-CO(2) (80:20) and sodium formate were not. The optimal growth temperature was 35 to 40 degrees C. The optimal pH range was 6.5 to 7.0. Both NaCl and Mg were required for growth, with maximum growth rates at 0.2 M NaCl and 0.05 M MgSO(4). The DNA base composition was 41 +/- 1% guanine plus cytosine. Methanosarcina acetivorans is the proposed species. C2A is the type strain (DSM 2834, ATCC 35395). PMID- 16346553 TI - Phage Resistance in a Phage-Insensitive Strain of Streptococcus lactis: Temperature-Dependent Phage Development and Host-Controlled Phage Replication. AB - Streptococcus lactis ME2 is a dairy starter strain that is insensitive to a variety of phage, including phi18. The efficiency of plating of phi18 on ME2 and N1 could be increased from <1 x 10 to 5.0 x 10 and from 7.6 x 10 to 2.1 x 10, respectively, when the host strains were subcultured at 40 degrees C before plating the phage and the phage assay plates were incubated at 40 degrees C. Host dependent replication was demonstrated in N1 at 30 degrees C and in N1 and ME2 at 40 degrees C, suggesting the operation of a temperature-sensitive restriction and modification system in ME2 and N1. The increased sensitivity of ME2 and N1 to phi18 at 40 degrees C was also demonstrated by lysis of broth cultures and increased plaque size. ME2 grown at 40 degrees C showed an increased ability to adsorb phi18, indicating a second target for temperature-dependent phage sensitivity in ME2. Challenge of N1 with a phi18 preparation that had been previously modified for growth on N1 indicated that at 40 degrees C phage development was characterized by a shorter latent period and larger burst size than at 30 degrees C. The evidence presented suggests that the high degree of phage insensitivity expressed by ME2 consists of a variety of temperature sensitive mechanisms, including (i) the prevention of phage adsorption, (ii) host controlled restriction of phage, and (iii) suppression of phage development. At 30 degrees C these factors appear to act cooperatively to prevent the successful emergence of lytic phage active against S. lactis ME2. PMID- 16346554 TI - Anaerobic biodegradation of the lignin and polysaccharide components of lignocellulose and synthetic lignin by sediment microflora. AB - Specifically radiolabeled [C-lignin]lignocelluloses and [C polysaccharide]lignocelluloses were prepared from a variety of marine and freshwater wetland plants including a grass, a sedge, a rush, and a hardwood. These [C]lignocellulose preparations and synthetic [C]lignin were incubated anaerobically with anoxic sediments collected from a salt marsh, a freshwater marsh, and a mangrove swamp. During long-term incubations lasting up to 300 days, the lignin and polysaccharide components of the lignocelluloses were slowly degraded anaerobically to CO(2) and CH(4). Lignocelluloses derived from herbaceous plants were degraded more rapidly than lignocellulose derived from the hardwood. After 294 days, 16.9% of the lignin component and 30.0% of the polysaccharide component of lignocellulose derived from the grass used (Spartina alterniflora) were degraded to gaseous end products. In contrast, after 246 days, only 1.5% of the lignin component and 4.1% of the polysaccharide component of lignocellulose derived from the hardwood used (Rhizophora mangle) were degraded to gaseous end products. Synthetic [C]lignin was degraded anaerobically faster than the lignin component of the hardwood lignocellulose; after 276 days, 3.7% of the synthetic lignin was degraded to gaseous end products. Contrary to previous reports, these results demonstrate that lignin and lignified plant tissues are biodegradable in the absence of oxygen. Although lignocelluloses are recalcitrant to anaerobic biodegradation, rates of degradation measured in aquatic sediments are significant and have important implications for the biospheric cycling of carbon from these abundant biopolymers. PMID- 16346555 TI - Effect of substrate concentration and organic and inorganic compounds on the occurrence and rate of mineralization and cometabolism. AB - Isopropyl N-phenylcarbamate (IPC) at 400 pg and 1 mug/ml was mineralized in samples of sewage, but only the lower concentration was mineralized in lake water samples in a 50-day period. IPC at 1 mug/ml disappeared from lake water, but it was converted to organic products. Mineralization of IPC at 400 pg/ml in lake water was enhanced by additions of inorganic nutrients or a mixture of nonchlorinated water pollutants but not by yeast extract or mixtures containing aromatic compounds or excretions of primary producers. The mineralization of 200 pg of 2,4-dichlorophenoxyacetate per ml of lake water was not affected by additions of low levels of yeast extract or compounds excreted by primary producers but was enhanced by low concentrations of mixtures of water pollutants. It is suggested that some chemicals that are found to be converted only to organic products, presumably by cometabolism, in tests using the concentrations commonly employed in laboratory evaluations may be mineralized at the lower concentrations prevailing in natural waters. PMID- 16346556 TI - Isolation of Yersinia ruckeri Bacteriophages. AB - Eight bacteriophages effective against Yersinia ruckeri, the enteric redmouth disease bacterium, were isolated. Phage YerA41, a tailed icosahedral virus isolated from sewage enrichments, lysed 34 of 35 strains of Y. ruckeri serovar I, but was inactive against 15 strains belonging to three other serological groups. Six other phages lysed strains of serovars II, V, and I', a subgroup of serovar I. YerL62, a phage obtained by mitomycin C induction, was specific for one of three serovar V strains. These bacteriophages, particularly YerA41, have potential value for fish disease diagnostic work. PMID- 16346557 TI - Selective isolation of acidophilic streptomyces strains for glucose isomerase production. AB - Approximately 260 Streptomyces strains were isolated from neutral pH farmland soil and evaluated for their ability to produce glucose isomerase. The number of acidophilic Streptomyces organisms growing at pH 4.0 was low, i.e., 10 organisms per g of soil. All of the isolates showed glucose isomerase activity when they were grown in a medium containing d-xylose, an inducer for glucose isomerase. More than half of the strains tested developed heavy growth in 24 h, and many produced high titers of glucose isomerase after 24 h of growth in a medium buffered at pH 5.0. PMID- 16346558 TI - Alcoholic Fermentation of d-Xylose by Yeasts. AB - Type strains of 200 species of yeasts able to ferment glucose and grow on xylose were screened for fermentation of d-xylose. In most of the strains tested, ethanol production was negligible. Nineteen were found to produce between 0.1 and 1.0 g of ethanol per liter. Strains of the following species produce more than 1 g of ethanol per liter in the fermentation test with 2% xylose: Brettanomyces naardenensis, Candida shehatae, Candida tenuis, Pachysolen tannophilus, Pichia segobiensis, and Pichia stipitis. Subsequent screening of these yeasts for their capacity to ferment d-cellobiose revealed that only Candida tenuis CBS 4435 was a good fermenter of both xylose and cellobiose under the test conditions used. PMID- 16346559 TI - Synthesis of Extracellular Proteinase by Pseudomonas fluorescens Under Conditions of Limiting Carbon, Nitrogen, and Phosphate. AB - The influence of carbon, nitrogen, and phosphate concentrations on growth and proteinase production by Pseudomonas fluorescens 32A was examined. In mineral salts medium containing dialyzed skim milk supernatant as an inducer, maximum growth was obtained at 1.0 and 2.5 mM orthophosphate at 20 and 5 degrees C, respectively. At both temperatures, 5 mM orthophosphate was required for maximum proteinase production, whereas significant inhibition was found at 10 mM. Orthophosphate was the only phosphate compound able to support growth. With sodium pyruvate as the carbon source, maximum enzyme synthesis was at 100 mM carbon at both temperatures. At both 20 and 5 degrees C maximum growth and enzyme production was found with 10 mM NH(4)Cl. A bioassay for available phosphate based on the growth of P. fluorescens 32A in phosphate-limited mineral salts medium showed that skim milk and skim milk supernatant contained 50 and 10 mM orthophosphate, respectively. Proteinase production in skim milk was 2.6- and 12 fold greater than that in optimal mineral salts medium at 20 and 5 degrees C, respectively. These results suggest that proteinase production in milk does not occur as a result of nutrient limitation and may be regulated in part by milk phosphates. PMID- 16346560 TI - Protection of Bacillus larvae from Oxygen Toxicity with Emphasis on the Role of Catalase. AB - Sporulation of Bacillus larvae NRRL B-3650 occurred only at aeration rates lower than those used for cultivation of most Bacillus species. One possible explanation for the requirement for a low level of aeration in B. larvae is that toxic forms of oxygen such as H(2)O(2) and superoxide are involved. The superoxide dismutase levels of strain B-3650 were similar to those of Bacillus subtilis 168 during sporulation, and no NADH peroxidase was present. Catalase activity was absent during exponential growth and first appeared near the start of the stationary phase. The catalase activity was 2,700 times less than that in B. subtilis 168 at the same stage of development. Therefore, the relative deficiency of catalase (and NADH peroxidase) might be the cause of the apparent O(2) toxicity. It was postulated that B. larvae might accumulate H(2)O(2) in the medium and exhibit more than normal sensitivity to H(2)O(2). Experimental results did not verify either postulate, but the possibilities of intracellular accumulation of H(2)O(2) and unusual sensitivity to endogenous H(2)O(2) were not excluded. The catalase present in early-stationary-phase cells was soluble, heat labile, and inhibited by cyanide, azide, and hydroxylamine. An increase in catalase activity also occurred at the time of appearance of refractile spores in both B. larvae NRRL B-3650 and B. subtilis 168. The level of catalase activity in strain B-3650 was 5,400 times less than that in B. subtilis 168 at this stage. In B. larvae, this second increase occurred primarily within the developing endospore. The activity in spore extracts was particulate, heat stable, and inhibited by hydroxylamine but not by azide or cyanide. Synthesis of catalase in B. larvae was unaffected by H(2)O(2), O(2), or glucose. PMID- 16346561 TI - Inhibition of alcoholic fermentation of grape must by Fatty acids produced by yeasts and their elimination by yeast ghosts. AB - In a complete nutritive medium rich in sugar, such as grape must, the inhibition of alcoholic fermentation is caused by substances produced by the yeast which, acting synergistically with ethanol, are toxic to the yeasts themselves. Among these are decanoic and octanoic acids and their corresponding ethyl esters. Their adsorption by yeast ghosts permits the prevention and treatment of fermentation stoppages. PMID- 16346562 TI - Use of gas-liquid chromatography to determine the end products of growth of lactic Acid bacteria. AB - A simple gas-liquid chromatographic procedure for analyzing ethanol, acetic acid, acetoin, and racemic and meso-2,3-butylene glycol in broth media is described. Overnight broth cultures were filtered or centrifuged, and the filtrate or supernatant was treated with formic acid to aid separation of volatile fatty acids. Samples were then directly analyzed by gas-liquid chromatography on a 20% Tween 80-Chromosorb W-AW column and propionic acid as an internal standard. A complete analysis took ca. 8 min. The method can be used to distinguish homofermentative from heterofermentative lactic acid bacteria based on the level of ethanol produced and citrate-utilizing from non-citrate-utilizing lactic acid bacteria based on the levels of acetic acid produced. The method also has potential in distinguishing other bacterial fermentations. Of the 13 species of lactic acid bacteria tested, Streptococcus lactis subsp. diacetylactis was the major producer of 2,3-butylene glycol (total range, 0.3 to 3.5 mM), and, except for strain DRC1, both the racemic and meso isomers were produced in approximately equal amounts. PMID- 16346563 TI - Seasonal study of methane oxidation in lake washington. AB - The distribution of methane and methane-oxidizing bacteria in the water column of Lake Washington was determined monthly for 1 year. The methane profiles were relatively constant, with little stratification and low concentrations (0.05 to 0.5 muM). The number of methane-oxidizing bacteria detected by a filter-plating method was routinely <1/ml throughout the water column, and no incorporation or oxidation of methane was detected by radioisotopic labeling, even after methane was added. However, samples taken from the sediment-water interface contained as much as 3 muM methane and 50 CFU of methane-oxidizing bacteria per ml and showed significant rates of methane oxidation and incorporation. To define the region of maximum activity more precisely, vertical profiles of the sediment were examined. The concentration of methane increased with depth to a maximum of 150 to 325 muM at 2.5 cm, and significant rates of methane oxidation were found within the top 2.5 cm. The apparent K(m)s for methane and oxygen were determined for samples from the top 1.0 cm of the sediment and found to be ca. 10 and 20 muM, respectively. Projected values for methane oxidation rates suggested that maximum methane oxidation occurred in the top 0.5 cm of the sediment. PMID- 16346564 TI - Agglutination Typing of Vibrio anguillarum Isolates from Diseased Fish and from the Environment. AB - Agglutinating activity was widely distributed among 101 Vibrio anguillarum strains of different origin and three Vibrio ordalii strains from salmonids. The spectrum of cells which were agglutinated comprised yeast cells and human (type O), poultry, guinea pig, and trout erythrocytes, whereas ovine, bovine, and tanned bovine erythrocytes were not affected. Mannose-sensitive hemagglutination, mannose-resistant hemagglutination, and non-agglutinating strains were recognized. The three V. ordalii strains showed mannose-resistant hemagglutination, whereas V. anguillarum exhibited either mannose-sensitive hemagglutination or was non-agglutinating. Among V. anguillarum, sensitivity to d galactose and l-fucose occurred sporadically. An agglutination typing scheme was developed for strains of V. anguillarum based on the agglutination pattern of human, poultry, guinea pig, and trout erythrocytes and yeast cells. Eight different agglutination types (A through H) were defined. The distribution of these types among fish pathogenic and environmental V. anguillarum strains were studied. The application of the typing scheme in ecological and epidemiological studies and for preventive medical purposes is discussed. PMID- 16346565 TI - Methane production in Minnesota peatlands. AB - Rates of methane production in Minnesota peats were studied. Surface (10- to 25 cm) peats produced an average of 228 nmol of CH(4) per g (dry weight) per h at 25 degrees C and ambient pH. Methanogenesis rates generally decreased with depth in ombrotrophic peats, but on occasion were observed to rise within deeper layers of certain fen peats. Methane production was temperature dependent, increasing with increasing temperature (4 to 30 degrees C), except in peats from deeper layers. Maximal methanogenesis from these deeper regions occurred at 12 degrees C. Methane production rates were also pH dependent. Two peats with pHs of 3.8 and 4.3 had an optimum rate of methane production at pH 6.0. The addition to peat of glucose and H(2)-CO(2) stimulated methanogenesis, whereas the addition of acetate inhibited methanogenesis. Cysteine-sulfide, nitrogen-phosphorus-trace metals, and vitamins-yeast extract affected methane production very little. Various gases were found to be trapped or dissolved (or both) within peatland waters. Dissolved methane increased linearly to a depth of 210 cm. The accumulation of metabolic end products produced within peat bogs appears to be an important mechanism limiting carbon turnover in peatland environments. PMID- 16346566 TI - Intermediary Metabolism in Clostridium acetobutylicum: Levels of Enzymes Involved in the Formation of Acetate and Butyrate. AB - The levels of seven intermediary enzymes involved in acetate and butyrate formation from acetyl coenzyme A in the saccharolytic anaerobe Clostridium acetobutylicum were investigated as a function of time in solvent-producing batch fermentations. Phosphate acetyltransferase and acetate kinase, which are known to form acetate from acetyl coenzyme A, both showed a decrease in specific activity when the organism reached the solvent formation stage. The three consecutive enzymes thiolase, beta-hydroxybutyrylcoenzyme A dehydrogenase, and crotonase exhibited a coordinate expression and a maximal activity after growth had ceased. Only low levels of butyryl coenzyme A dehydrogenase activity were found. Phosphate butyryltransferase activity rapidly decreased after 20 h from 5 to 11 U/mg of protein to below the detection limit (1 mU/mg). Butyrate no longer can be formed, and the metabolic flux may be diverted to butanol. Butyrate kinase showed a 2.5- to 10-fold increase in specific activity after phosphate butyryltransferase activity no longer could be detected. These results suggest that the uptake of acetate and butyrate during solvent formation can not proceed via a complete reversal of the phosphate transferase and kinase reactions. The activities of all enzymes investigated as a function of time in vitro are much higher than the metabolic fluxes through them in vivo. This indicates that none of the maximal activities of the enzymes assayed is rate limiting in C. acetobutylicum. PMID- 16346567 TI - Production of nitric oxide in loam under aerobic and anaerobic conditions. AB - Measurements of gas flow through soil columns of loam from Kjettslinge, Uppland, Sweden, gave average NO production rates of 0.06 +/- 0.01 ng of NO N g of soil min in aerobic conditions and 3.7 +/- 0.6 ng of NO N g of soil min in anaerobic conditions at 25 degrees C. Approximately 30% of the NO(3) loss in anaerobic conditions was as NO. In aerobic conditions an equilibrium concentration for NO was found. Above this concentration there was uptake of NO. Autoclaved samples indicated that less than 10% of the NO production was abiological, and there was no abiological NO uptake. The NO production reached anaerobic rates at soil O(2) levels between 0.5 and 0.05%. PMID- 16346568 TI - Identification of nitroso compounds from biotransformation of 2,4-dinitrotoluene. AB - The intermediates of microbial transformation of 2,4-dinitrotoluene by a mixed bacterial culture derived from activated sludge were identified as 2-amino-4 nitrotoluene, 4-amino-2-nitrotoluene, 2-nitroso-4-nitrotoluene, and 4-nitroso-2 nitrotoluene. The biotransformation of 2,4-dinitrotoluene occurred only under anaerobic conditions with an exogenous carbon source. The two nitroso compounds were unstable and could be observed only at the early stage of 2,4-dinitrotoluene anaerobic biotransformation. PMID- 16346569 TI - Separation of fungal propagules by partition in aqueous polymer two-phase systems. AB - Conidia of Penicillium chrysogenum and Penicillium frequentans and sporangiospores of Rhizopus rhizopodiformis, Rhizomucor pusillus, and Mucor racemosus were subjected to partition in aqueous polymer two-phase systems. The partition behavior differed drastically between the conidia of the two Penicillium species and the sporangiospores of the three species of phycomycetes. This difference in partition behavior can be used for purification of fungi belonging to different taxonomical groups. P. frequentans was completely separated from M. racemosus by two extractions, whereas four extractions were needed to purify M. racemosus. This method was used on an air sample from a locality where wood fuel chips are handled. The conidia of the fungi Trichoderma viride and Rhizopus rhizopodiformis were removed completely by only two extractions. PMID- 16346570 TI - Pigment Production from Immobilized Monascus sp. Utilizing Polymeric Resin Adsorption. AB - Pigment production by the fungus Monascus sp. was studied to determine why Monascus sp. provides more pigment in solid culture than in submerged culture. Adding a sterilized nonionic polymeric adsorbent resin directly to the growing submerged culture did not enhance the pigment production, thus indicating that pigment extraction is probably not a factor. Monascus cells immobilized in hydrogel were studied and exhibited decreased pigment production as a result of immobilization. This result is thought to be due to diffusional resistance of the pigment through the hydrogel beads. Addition of the adsorbent resin to the immobilized Monascus culture increased both the maximum pigment yield and the production rate above those of the free-cell fermentations. The provision of a support for the mycelium may explain enhanced pigment production by the solid state culture. These results indicate that product diffusion from immobilized cell systems can be the limiting factor and that in situ extraction is one possible way to circumvent this problem. PMID- 16346571 TI - Responses of Sorghum and Pennisetum Species to the N(2)-Fixing Bacterium Azospirillum brasilense. AB - Three field inoculation experiments, two in Florida and one in New Mexico, were conducted with Azospirillum brasilense Cd. Each of the Florida experiments evaluated two crop species. One species in each of the Florida experiments responded to inoculation with a significant dry matter yield increases of 11 to 24% and nitrogen yield increases of 9 to 39%. No inoculation response was noted in the New Mexico experiment. The responding species were Sorghum bicolor (L.) Moench (sorghum) and the interspecific hybrid between Pennisetum americanum (L.) K. Schum. (pearl millet) and P. purpureum Schumach. (napiergrass). Nonresponding species were pearl millet (Florida) and Sorghum sudanense (Piper) Staph. (New Mexico). Acetylene reduction activity of inoculated plots in Florida was low, showing no increase over the natural uninoculated background rates and, in one case, was negatively correlated with yield. Acetylene reduction activity was not measured in New Mexico. In Florida, A. brasilense populations were found to decline from 5 x 10 to 5 x 10 bacteria g of soil in about 3 weeks (quadratic regressions). Continued decline to less than 10 by week 5 indicated that the inoculated bacteria did not become established in the soil in high numbers. The A. brasilense population declined at about the same rate in the New Mexico experiment. The erractic inoculation responses in these experiments are similar to those observed in earlier work at the University of Florida. The lack of acetylene reduction activity response to inoculation and the rapid population decline of the inoculated bacteria suggest that N(2) fixation is not the major mechanism causing yield responses after inoculation. PMID- 16346572 TI - Selective inhibition by 2-bromoethanesulfonate of methanogenesis from acetate in a thermophilic anaerobic digestor. AB - The effects of 2-bromoethanesulfonate, an inhibitor of methanogenesis, on metabolism in sludge from a thermophilic (58 degrees C) anaerobic digestor were studied. It was found from short-term experiments that 1 mumol of 2 bromoethanesulfonate per ml completely inhibited methanogenesis from CH(3)COO, whereas 50 mumol/ml was required for complete inhibition of CO(2) reduction. When 1 mumol of 2-bromoethanesulfonate per ml was added to actively metabolizing sludge which was then incubated for 24 h. it caused a 60% reduction in methanogenesis and a corresponding increase in acetate accumulation; at 50 mumol/ml it caused complete inhibition of methanogenesis and accumulation of acetate. H(2), and ethanol. PMID- 16346573 TI - Isolation of Streptococcus lactis Bacteriophages and Their Interaction with the Host Cell. AB - Phages may cause lysis of lactic acid bacteria used in cheese production. Three virulent bacteriophages specific for Streptococcus lactis subsp. lactis C2 were isolated and purified from cheese whey. They showed distinct plaque sizes, and although they had similar morphology by electron microscope examination, their dimensions were slightly different. The phage heads were elongated and hexagonal in shape, and the flexible tails appeared periodically cross-striated. They were DNA phages based on the acridine orange test. On infection, phage was adsorbed on the bacterial surface by the free end of the tail. After 80 min of incubation at 25 degrees C, the phage heads appeared empty, slightly collapsed, and possessed a visible hollow tube through which the genetic material had been injected. PMID- 16346574 TI - Role of N-Acetylgalactosamine-Specific Protein in Trapping of Nematodes by Arthrobotrys oligospora. AB - An N-acetylgalactosamine-specific protein was purified from mycelial homogenates of the nematode-trapping fungus Arthrobotrys oligospora by using affinity chromatography. The molecular weight of the protein was estimated at 22,000 by its comparative mobility on sodium dodecyl sulfate-polyacrylamide slab gels. Pretreatment of nematodes with the purified protein reduced entrapment, indicating a role for the sugar-binding protein in recognition and capture of prey by the fungus. PMID- 16346576 TI - Factors determining annual changes in bacterial photosynthetic pigments in holomictic lake ciso, Spain. AB - [This corrects the article on p. 1000 in vol. 46.]. PMID- 16346575 TI - Anaerobic degradation of coniferyl alcohol by methanogenic consortia. AB - [This corrects the article on p. 1445 in vol. 46.]. PMID- 16346577 TI - Sustained Photoproduction of Ammonia from Dinitrogen and Water by the Nitrogen Fixing Cyanobacterium Anabaena sp. Strain ATCC 33047. AB - Conditions have been developed that lengthen the time during which photosynthetic dinitrogen fixation by filaments of the cyanobacterium Anabaena sp. strain ATCC 33047 proceeds freely, whereas the subsequent conversion of ammonia into organic nitrogen remains blocked, with the resulting ammonia released to the outer medium. When l-methionine-dl-sulfoximine was added every 20 h, maximal rates of ammonia production (25 to 30 mumol/mg of chlorophyll per h) were maintained for about 50 h. After this time, ammonia production ceased due to a deficiency of glutamine and other nitrogenous compounds in the filaments, conditions which finally led to cell lysis. The effective ammonia production period could be further extended to about 7 days by adding a small amount of glutamine at the end of a 40-h production period or by allowing the cells to recover for 8 h in the absence of l-methionine-dl-sulfoximine after every 40-h period in the presence of the inhibitor. A more prolonged steady production of ammonia, lasting for longer than 2 weeks, was achieved by alternating treatments with the glutamine synthetase inhibitors l-methionine-dl-sulfoximine and phosphinothricin, provided that 8-h recovery periods in the absence of either compound were also alternated throughout. The biochemically manipulated cyanobacterial filaments thus represent a system that is relatively stable with time for the conversion of light energy into chemical energy, with the net generation of a valuable fuel and fertilizer through the photoreduction of dinitrogen to ammonia. PMID- 16346578 TI - Mixed-culture fermentor for simulating methanogenic digestors. AB - Propionate degradation in an anaerobic digestor degrading animal waste (10-day retention time, 5.75 g liter day volatile solids loading rate, 40 degrees C) was 0.304 mM h, measured with [2-C]propionate; this value indicated that CH(4) produced from propionate accounted for 14.8% of the CH(4) produced in the digestor (34.5%, including acetate produced from propionate). The mean propionate concentration was 0.67 mM, giving a propionate turnover rate of 0.46 h. A continuous-, mixed-culture fermentor was developed to mimic the digestor. When degradation rates of methanogenic precursors (H(2), CO(2), and acetate) equalled those measured in the digestor, propionate degradation was inhibited. When the H(2) turnover rate was lowered by decreasing addition of H(2)-generating substrates or by allowing a portion of the H(2) degradation to occur in an isolated compartment, propionate degradation in the fermentor resumed. The possibility is discussed that in digestors, much of the H(2) is produced and degraded within microenvironments associated with particles. Thus, the gross turnover rate of H(2) measured in digestors is an average, and specific microenvironments within the digestor may have different rates of turnover. PMID- 16346579 TI - Light and electron microscopic examinations of methane-producing biofilms from anaerobic fixed-bed reactors. AB - Ultrastructural examinations were performed on biofilms from eight anaerobic fixed-bed reactors filled with various packing materials and operated on fresh swine waste. By using light, UV, scanning, and transmission electron microscopy, the distribution of a diverse microbial population composed of bacteria and a few yeasts was determined. This is the first time that the ultrastructure of in situ anaerobic digestor biofilms has been reported. A large number of methanogenic bacteria were identified by their fluorescence under 420 nm of radiation. Of these, two morphologically distinct types were most prevalent in the films. Methanothrix spp. was present in high numbers at the film surface, whereas Methanosarcina spp. were commonly embedded in the lower regions of the film. Inhabitants of the film were surrounded by an exopolysaccharide matrix that was very dense toward the base. An extensive network of channels was observed throughout the matrix that may facilitate gas and nutrient exchange to the lower regions of the film. PMID- 16346580 TI - Propagation of Rhizopus javanicus Biosorbent. AB - After propagation of Rhizopus javanicus in defined media containing glucose, urea, and mineral salts in deionized distilled water, the ability of the nonliving biomass to sequester cupric ion was assayed. Growth, uptake capacity (saturation uptake at >1 mM Cu concentration in solution), and biosorptive yield (biomass concentration x uptake capacity) were increased by augmentation of the growth medium with mineral salts once growth was under way. In the stationary phase, the uptake capacity of mycelia, which were normally a poor biosorbent, was improved within 4 h of trace metal addition to the growth medium. Growth of the culture was inhibited by excessive concentrations (0.04 to 40 muM) of metals in the medium in the following order: Cu > Co >/= Ni > Mn > Mo; zinc was not inhibitory at 40 muM, and chromium was stimulatory at 0.53 muM but slightly inhibitory at higher levels. Iron and potassium phosphate stimulated growth at levels of 0.53 and 40 mM, respectively. When R. javanicus was propagated in a medium with a high salt concentration, exponential growth (0.23 h) to a biomass concentration of >3 g/liter and a biosorptive yield of >500 mumol/liter was achieved. It is evident that the powerful biosorbent characteristics of Rhizopus biomass led to depletion of available trace minerals in suspension culture, which in turn limited growth. PMID- 16346581 TI - Evolution of acetic Acid bacteria during fermentation and storage of wine. AB - Acetic acid bacteria were present at all stages of wine making, from the mature grape through vinification to conservation. A succession of Gluconobacter oxydans, Acetobacter pasteurianus, and Acetobacter aceti during the course of these stages was noted. Low levels of A. aceti remained in the wine; they exhibited rapid proliferation on short exposure of the wine to air and caused significant increases in the concentration of acetic acid. Higher temperature of wine storage and higher wine pH favored the development and metabolism of these species. PMID- 16346582 TI - Size of suspended bacterial cells and association of heterotrophic activity with size fractions of particles in estuarine and coastal waters. AB - The size of bacteria and the size distribution of heterotrophic activity were examined in estuarine, neritic, and coastal waters. The data indicated the small size of suspended marine bacteria and the predominance of free-living cells in numerical abundance and in the incorporation of dissolved amino acids. The average per-cell volume of suspended marine bacteria in all environments was less than 0.1 mum. Cell volume ranged from 0.072 to 0.096 mum at salinities of 0 to 34.3 per thousand in the Newport River estuary, N.C., and from 0.078 to 0.096 mum in diverse areas of the Gulf of Mexico. Thus, the free-living bacteria were too small to be susceptible to predation by copepods. In the Newport River estuary, ca. 93 to 99% of the total number of cells and 75 to 97% of incorporated tritium (from H-labeled mixed amino acids) retained by a 0.2-mum-pore-size filter passed through a 3.0-mum-pore-size filter. Although the amino acid turnover rate per cell was higher for the bacteria in the >3.0-mum size fraction than in the <3.0 mum size fraction, the small number of bacteria associated with the >3.0-mum size particles resulted in the low relative contribution of attached bacteria to total heterotrophic activity in the estuary. For coastal and neritic samples, collected off the coast of Georgia and northeast Florida and in the plume of the Mississippi River, 56 to 98% of incorporated label passed through a 3.0-mum-pore size filter. The greatest activity in the >3.0-mum fraction in the Georgia Bight was at nearshore stations and in the bottom samples. Our data were consistent with the hypothesis that resuspension of bottom material is an important factor in influencing the proportion of heterotrophic activity attributable to particle associated bacteria. PMID- 16346583 TI - Diel production and microheterotrophic utilization of dissolved free amino acids in waters off southern california. AB - Diel patterns of dissolved free amino acid (DFAA) concentration and microheterotrophic utilization were examined in the spring and fall of 1981 in euphotic waters from the base of the mixed layer off the southern California coast. The average depths of the isotherms sampled were 19.2 m for spring and 9.0 m for fall. Total DFAA levels were generally higher in the spring than in the fall, 18 to 66 nM and 14 to 20 nM, respectively. Two daily concentration maxima and minima were observed for total DFAAs as well as for most individual DFAAs. Maxima were usually measured in the mid-dark period and in the early afternoon; minima were typically observed in early morning and late afternoon. Bacterial cell numbers reached maximal values near midnight in both seasons. These increases coincided with one of the total DFAA maxima. The second total DFAA maximum occurred in early to midafternoon, during the time of maximum photosynthetic carbon production and rapid dissolved amino acid utilization. Microbial metabolism (incorporation plus respiration) of selected H-amino acids was 2.7 to 4.1 times greater during the daylight hours. DFAA turnover times, based on these metabolic measurements, ranged between 11 and 36 h for the amino acids tested, and rates were 1.7 to 3.7 times faster in the daylight hours than at night. DFAA distributions were related to primary production and chlorophyll a concentrations. Amino acids were estimated to represent 9 to 45% of the total phytoplankton exudate. Microheterotrophic utilization or production of total protein amino acids was estimated as 3.6 mug of C liter day in spring and 1.9 mug of C liter day in the fall. Assimilation efficiency for dissolved amino acids averaged 65% for marine microheterotrophs. PMID- 16346584 TI - Development of Rapidly Fermenting Strains of Saccharomyces diastaticus for Direct Conversion of Starch and Dextrins to Ethanol. AB - Alcoholic fermentation, growth, and glucoamylase production by 12 strains of Saccharomyces diastaticus were compared by using starch and dextrins as substrates. Haploid progeny produced from a rapidly fermenting strain, SD2, were used for hybridization with other S. diastaticus and Saccharomyces cerevisiae haploids. Alcoholic fermentation and enzyme production by hybrid diploids and their haploid parents were evaluated. Although the dosage of the STA or DEX (starch or dextrin fermentation) genes may enhance ethanol production, epistatic effects in certain strain combinations caused decreases in starch-fermenting activity. Both the nature of the starch or dextrin used and the fermentation medium pH had substantial effects on alcohol production. Commercial dextrin was not as good a substrate as dextrins prepared by digesting starch with alpha amylase. Crude manioc starch digested by alpha-amylase was fermented directly by selected hybrids with almost 100% conversion efficiency. The manioc preparation contained adequate minerals and growth factors. This procedure should be suitable for direct commercial application in manioc-producing regions in Brazil and elsewhere. A rapidly fermenting haploid strain, SD2-A8, descended from strain SD2, contains two unlinked genes controlling formation of extracellular amylase. A convenient method for detecting these genes (STA genes) in replica plates containing large numbers of meiotic progeny was developed. PMID- 16346585 TI - Ethanol Production by Thermophilic Bacteria: Physiological Comparison of Solvent Effects on Parent and Alcohol-Tolerant Strains of Clostridium thermohydrosulfuricum. AB - The effects of temperature, solvents, and cultural conditions on the fermentative physiology of an ethanol-tolerant (56 g/liter at 60 degrees C) and parent strain of Clostridium thermohydrosulfuricum were compared. An ethanol-tolerant mutant was selected by successive transfer of the parent strain into media with progressively higher ethanol concentrations. Physiological differences noted in the mutant included enhanced growth, tolerance to various solvents, and alterations in the substrate range and the fermentation end product ratio. Ethanol tolerance was temperature dependent in the mutant but not in the parent strain. The mutant grew with ethanol concentrations up to 8.0% (wt/vol) at 45 degrees C, but only up to 3.3% (wt/vol) at 68 degrees C. Low ethanol concentration (0.2 to 1.6% [wt/vol]) progressively inhibited the parent strain to where glucose was not fermented at 2.0% (wt/vol) ethanol. Both strains grew and produced alcohols on glucose complex medium at 60 degrees C in the presence of either 5% methanol or acetone, and these solvents when added at low concentration stimulated fermentative metabolism. The mutant produced ethanol at high concentrations and displayed an ethanol/glucose ratio (mole/mole) of 1.0 in media where initial ethanol concentrations were 210 > 250. Results of in vitro studies concerning tolerance responses of ectomycorrhizal fungi to aluminum and manganese were not consistent with field observations of the successional sequence of these fungi on acid coal spoils. PMID- 16346624 TI - Formation of n-Butanol from d-Glucose by Strains of the "Clostridium tetanomorphum" Group. AB - A clostridial strain has been isolated that produced n-butanol, ethanol, butyrate, and acetate as major fermentation products from glucose but no acetone. At a pH of 6.6, n-butanol was formed by this microorganism only during growth. On the basis of its physiological characteristics and DNA-DNA homology data, the strain was assigned to the "Clostridium tetanomorphum" group (S. Nakamura, I. Okado, T. Abe, and S. Nishida, J. Gen. Microbiol. 113:29-35, 1979). All members of this group were shown to produce n-butanol from glucose as the major fermentation product, whereas C. cochlearium produced it in only minor amounts. PMID- 16346625 TI - Ethanol from Whey: Continuous Fermentation with a Catabolite Repression-Resistant Saccharomyces cerevisiae Mutant. AB - An alternative method for the conversion of cheese whey lactose into ethanol has been demonstrated. With the help of continuous-culture technology, a catabolite repression-resistant mutant of Saccharomyces cerevisiae completely fermented equimolar mixtures of glucose and galactose into ethanol. The first step in this process was a computer-controlled fed-batch operation based on the carbon dioxide evolution rate of the culture. In the absence of inhibitory ethanol concentrations, this step allowed us to obtain high biomass concentrations before continuous fermentation. The continuous anaerobic process successfully incorporated a cell-recycle system to optimize the fermentor productivity. Under conditions permitting a low residual sugar concentration (85% of the total population was found to be active by at least one direct activity measurement. Thus, active rather than dormant organisms compose a major portion of the microbial population in this region of Chesapeake Bay. PMID- 16346660 TI - Degradation of 4-Chlorobenzoic Acid by Arthrobacter sp. AB - A mixed population, enriched and established in a defined medium, from a sewage sludge inoculum was capable of complete mineralization of 4-chlorobenzoate. An organism, identified as Arthrobacter sp., was isolated from the consortium and shown to be capable of utilizing 4-chlorobenzoate as the sole carbon and energy source in pure culture. This organism (strain TM-1), dehalogenated 4 chlorobenzoate as the initial step in the degradative pathway. The product, 4 hydroxybenzoate, was further metabolized via protocatechuate. The ability of strain TM-1 to degrade 4-chlorobenzoate in liquid medium at 25 degrees C was improved by the use of continuous culture and repeated sequential subculturing. Other chlorinated benzoates and the parent compound benzoate did not support growth of strain TM-1. An active cell extract was prepared and shown to dehalogenate 4-chloro-, 4-fluoro-, and 4-bromobenzoate. Dehalogenase activity had an optimum pH of 6.8 and an optimum temperature of 20 degrees C and was inhibited by dissolved oxygen and stimulated by manganese (Mn). Strain improvement resulted in an increase in the specific activity of the cell extract from 0.09 to 0.85 nmol of 4-hydroxybenzoate per min per mg of protein and a decrease in the doubling time of the organism from 50 to 1.6 h. PMID- 16346661 TI - Evolution of yeasts and lactic Acid bacteria during fermentation and storage of bordeaux wines. AB - The levels of yeasts and lactic acid bacteria that naturally developed during the vinification of two red and two white Bordeaux wines were quantitatively examined. Yeasts of the genera Rhodotorula, Pichia, Candida, and Metschnikowia occurred at low levels in freshly extracted grape musts but died off as soon as fermentation commenced. Kloeckera apiculata (Hanseniaspora uvarum), Torulopsis stellata, and Saccharomyces cerevisiae, the dominant yeasts in musts, proliferated to conduct alcoholic fermentation. K. apiculata and eventually T. stellata died off as fermentation progressed, leaving S. cerevisiae as the dominant yeast until the termination of fermentation by the addition of sulfur dioxide. At least two different strains of S. cerevisiae were involved in the fermentation of one of the red wines. Low levels of lactic acid bacteria (Pediococcus cerevisiae, Leuconostoc mesenteroides, and Lactobacillus spp.) were present in grape musts but died off during alcoholic fermentation. The malolactic fermentation developed in both red wines soon after alcoholic fermentation and correlated with the vigorous growth of at least three different strains of Leuconostoc oenos. PMID- 16346662 TI - Phosphonate Utilization by the Glyphosate-Degrading Pseudomonas sp. Strain PG2982. AB - The glyphosate-degrading Pseudomonas sp. strain PG2982 was found to utilize each of 10 organophosphonate compounds as a sole phosphorus source. Representative compounds tested included alkylphosphonates, 1-amino-substituted alkylphosphonates, amino-terminal phosphonates, and an arylphosphonate. This report demonstrates that PG2982 is capable of utilizing a wider range of structurally different organophosphonate compounds than any organism described to date. PMID- 16346663 TI - Production of d-Tagatose from Dulcitol by Arthrobacter globiformis. AB - A process for the bacterial oxidation of dulcitol to d-tagatose has been developed. The strain Arthrobacter globiformis ST48 used in this fermentation was isolated from soil. The yield of d-tagatose accumulated in the medium from dulcitol was as high as 85%. About 14 g of d-tagatose crystals was isolated from 1 liter of 2% dulcitol medium. PMID- 16346664 TI - Anaerobic biodegradation of indole to methane. AB - Methane gas was produced from a laboratory, granular activated carbon, anaerobic filter treating a synthetically prepared mixture of polycyclic N-aromatic compounds. The biodegradability of the individual polycyclic N-aromatic compound present in the mixture was investigated. Experimental results obtained from test bottles containing methanogenic enrichment cultures suggested that indole was degraded to methane and carbon dioxide under strict anaerobic conditions. PMID- 16346665 TI - Selection of an Asporogenous Strain of Clostridium acetobutylicum in Continuous Culture Under Phosphate Limitation. AB - Based on the observation that cells of Clostridium acetobutylicum unable to store granulose do not initiate sporulation, a staining procedure was developed for the detection of asporogenous mutants. By application of this procedure it was shown that an asporogenous strain of C. acetobutylicum was selected in continuous culture under phosphate limitation. PMID- 16346668 TI - Comparison of p-Nitrophenol Biodegradation in Field and Laboratory Test Systems. AB - Acclimation of microbial communities exposed to p-nitrophenol (PNP) was measured in laboratory test systems and in a freshwater pond. Laboratory tests were conducted in shake flasks with water, shake flasks with water and sediment, eco cores, and two sizes of microcosm. The sediment and water samples used in the laboratory experiments were obtained from the pond. After a 6-day acclimation period, PNP was biodegraded rapidly in the pond. When the pond was treated with PNP a second time, biodegradation began immediately. The acclimation periods in laboratory test systems that contained sediment were similar to that in the pond. The acclimation period was threefold longer in shake flasks without sediment. PNP was biodegraded more slowly by microbial communities acclimated in the laboratory than it was in the pond, and the rate of biodegradation varied with the type of test. The number of bacteria able to mineralize PNP increased by 3 orders of magnitude in the pond during the acclimation period. Similar increases accompanied acclimation in the laboratory systems. PMID- 16346669 TI - Multilocus Electrophoretic Assessment of the Genetic Structure and Diversity of Yersinia ruckeri. AB - Multilocus isoenzyme electrophoresis was used to screen 47 field isolates of Yersinia ruckeri for electrophoretic variation at 15 enzyme loci. Only four electrophoretic types were observed, thus indicating that the genetic structure of Y. ruckeri is clonal. Forty-two isolates were of one electrophoretic type, a reflection of the low amount of genetic diversity extant in this species. Although sorbitol fermentation has been considered to be indicative of a second biotype, no significant gene frequency differences were found between the group of 20 isolates that readily used sorbitol as the sole carbon source and the group of 27 that did not. PMID- 16346670 TI - Production and Regeneration of Lactobacillus casei Protoplasts. AB - Methods for the production and regeneration of Lactobacillus casei protoplasts are described. Protoplasts of L. casei strains were obtained by treatment with mutanolysin or with mutanolysin and lysozyme together in a protoplast formation buffer containing 0.02 M HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) (pH 7.0), 1 mM MgCl(2), 0.5% gelatin, and 0.3 M raffinose. Cells were regenerated on a complex medium supplemented with bovine serum albumin, MgCl(2), CaCl(2), gelatin, and raffinose. Lengthy digestion with lytic enzymes inhibited the capacity of protoplasts to regenerate. The optimum conditions of protoplast formation varied from strain to strain. Using predetermined optimal conditions it was possible to prepare protoplasts of several L. casei strains and regenerate them with 10 to 40% efficiency. The methods were applicable to other species of lactobacilli as well. PMID- 16346671 TI - Germination of Rhizopus oligosporus Sporangiospores. AB - The morphology of Rhizopus oligosporus (NRRL 2710) sporangiospores and their physiological requirements for germination were studied. Germination proceeded in two separable phases: phase I (swelling) and phase II (germ tube protrusion). The optimal conditions for germination were 42 degrees C and pH 4.0. Sporangiospores contained insufficient endogenous carbon for swelling or germination to occur in distilled water. Initial swelling during phase I occurred only in the presence of a suitable carbohydrate. Subsequent production of germ tubes during phase II required exogenous sources of both carbon and nitrogen. Spores germinated most rapidly in mixtures of amino acids; l-proline and l-alanine were the most effective. These amino acids, at concentrations as low as 10 M, supported germination when combined with glucose and McIlvaine (citric acid-phosphate) buffer. d-Glucose, d-xylose, and d-mannose were the most effective carbohydrates tested for promotion of germination. PMID- 16346672 TI - Influence of pH on Terminal Carbon Metabolism in Anoxic Sediments from a Mildly Acidic Lake. AB - The carbon and electron flow pathways and the bacterial populations responsible for transformation of H(2)-CO(2), formate, methanol, methylamine, acetate, glycine, ethanol, and lactate were examined in sediments collected from Knaack Lake, Wis. The sediments were 60% organic matter (pH 6.2) and did not display detectable sulfate-reducing activity, but they contained the following average concentration (in micromoles per liter of sediment) of metabolites and end products: sulfide, 10; methane, 1,540; CO(2), 3,950; formate, 25; acetate, 157; ethanol, 174; and lactate, 138. Methane was produced predominately from acetate, and only 4% of the total CH(4) was derived from CO(2). Methanogenesis was limited by low environmental temperature and sulfide levels and more importantly by low pH. Increasing in vitro pH to neutral values enhanced total methane production rates and the percentage of CO(2) transformed to methane but did not alter the amount of CO(2) produced from [2-C]acetate ( approximately 24%). Analysis of both carbon transformation parameters with C-labeled tracers and bacterial trophic group enumerations indicated that methanogenesis from acetate and both heterolactic- and acetic acid-producing fermentations were important to the anaerobic digestion process. PMID- 16346673 TI - Bicarbonate Uptake by Nitrifiers: Effects of Growth Rate, pH, Substrate Concentration, and Metabolic Inhibitors. AB - The ratios of bicarbonate uptake to substrate oxidation were measured for three genera of nitrifying bacteria. The ratios for the two ammonium oxidizers tested were essentially the same; 0.0863 +/- 0.0055 and 0.0868 +/- 0.0091 mumol of bicarbonate were taken up per umol ammonium oxidized for Nitrosomonas europaea and a Nitrosospira strain, respectively. For Nitrobacter sp., a ratio of 0.0236 +/- 0.0013 mumol of bicarbonate taken up per umol of nitrite oxidized was obtained. Cells were grown in substrate-limited continuous culture and in batch culture, with generation times ranging between 16 and 189 h for the ammonium oxidizers and 18 and 69 h for Nitrobacter sp. All ratios appeared to be independent of growth rates and pH. However, short-term changes in substrate concentration and certain metabolic inhibitors significantly changed the efficiency of bicarbonate uptake. The significance of these results to the application of the nitrapyrin-sensitive bicarbonate uptake method for measuring nitrification rates in natural samples is discussed. PMID- 16346674 TI - The Proteolytic Systems of Streptococcus cremoris: an Immunological Analysis. AB - The cell-wall-associated proteolytic systems of several Streptococcus cremoris strains were analyzed by crossed immunoelectrophoresis. At least four immunologically different components of the proteolytic system's were found. One of these proteins was produced by all strains tested. The proteolytic activity of this enzyme was demonstrated with a zymogram staining technique which is based on the degradation of Coomassie-brilliant-blue-stainable casein. The crossed immunoelectrophoresis patterns of the proteolytic systems of different S. cremoris strains indicated that each strain produces a characteristic combination of proteins. On the basis of these combinations, the different S. cremoris strains were classified into four groups. PMID- 16346675 TI - Production of Coenzyme A by a Mutant of Brevibacterium ammoniagenes Resistant to Oxypantetheine. AB - For improved production of coenzyme A (CoA), a mutant of Brevibacterium ammoniagenes IFO127071 resistant to oxypantetheine, the corresponding oxygen analog of pantetheine, was obtained. In the mutant, activity of pantothenate kinase (EC 2.7.1.33), the first-step enzyme for the biosynthesis of CoA from pantothenic acid, l-cysteine, and ATP, was about threefold higher than that in the parent strain. As the main regulation mechanism of CoA biosynthesis in this bacterium is negative feedback inhibition of pantothenate kinase by CoA, the mutant is very useful as a catalyst for practical production of CoA. When added to culture broth of the mutant, pantothenate, l-cysteine, and AMP gave 9.3 mg of CoA per ml. With pantetheine and AMP, 11.5 mg of CoA per ml accumulated. These values were about threefold higher than those with the parent strain, and more than 70% of the added AMP was converted to CoA. PMID- 16346676 TI - Isolation and characterization of a novel thermophilic, freshwater methanogen. AB - A novel thermophilic, coccoid methanogen isolated from nonthermal freshwater sediments is described. Hydrogen plus carbon dioxide and formate were substrates for methanogenesis, and methane production was stimulated by yeast extract, Casamino Acids, and tryptose. Growth also occurred autotrophically. Elevated levels of sodium chloride were not required for maximum growth and were inhibitory above 2%. The minimum doubling time occurred at 57 degrees C, and the upper and lower limits for methane production were 62 and 26 degrees C, respectively. The optimum pH for growth was between 7.0 and 7.5. Inhibitory antibiotics included metronidazole, anisomycin, chloramphenicol, and lasalocid. Colonies were circular, dark yellow, shiny, and convex with entire edges. Cells were 1 to 2.5 mum in diameter, nonmotile, occurring singly or in pairs, and fimbriated. Cells were lysed by pronase or trypsin digestion, glass-distilled water, and 1% sodium dodecyl sulfate. Electron micrographs of thin sections showed a monolayered cell wall ca. 20 nm thick. The DNA base ratio was 49.2 mol% guanine plus cytosine. The whole cell protein pattern differed from that of other named coccoid methanogens. PMID- 16346677 TI - Uncoupling by Acetic Acid Limits Growth of and Acetogenesis by Clostridium thermoaceticum. AB - When cells of the anaerobic thermophile Clostridium thermoaceticum grow in batch culture and homoferment glucose to acetic acid, the pH of the medium decreases until growth and then acid production cease, at about pH 5. We postulated that the end product of fermentation limits growth by acting as an uncoupling agent. Thus, when the pH of the medium is low, the cytoplasm of the cells becomes acidified below a tolerable pH. We have therefore measured the internal pH of growing cells and compared these values with those of nongrowing cells incubated in the absence of acetic acid. Growing cells maintained an interior about 0.6 pH units more alkaline than the exterior throughout most of batch growth (i.e., DeltapH = 0.6). We also measured the transmembrane electrical potential (DeltaPsi), which decreased from 140 mV at pH 7 at the beginning of growth to 80 mV when the medium had reached pH 5. The proton motive force, therefore, was 155 mV at pH 7, decreasing to 120 mV at pH 5. When further fermentation acidified the medium below pH 5, both the DeltapH and the DeltaPsi collapsed, indicating that these cells require an internal pH of at least 5.5 to 5.7. Cells harvested from stationary phase and suspended in citrate-phosphate buffer maintained a DeltapH of 1.5 at external pH 5.0. This DeltapH was dissipated by acetic acid (at the concentrations found in the growth medium) and other weak organic acids, as well as by ionophores and inhibitors of glycolysis and of the H-ATPase. Nongrowing cells had a DeltaPsi which ranged from about 116 mV at external pH 7 to about 55 mV at external pH 5 and which also was sensitive to ionophores. Since acetic acid, in its un-ionized form, diffuses passively across the cytoplasmic membrane, it effectively renders the membrane permeable to protons. It therefore seems unlikely that mutations at one or a few loci would result in C. thermoaceticum cells significantly more acetic acid tolerant than their parental type. PMID- 16346678 TI - Production of Solvents by Clostridium acetobutylicum Cultures Maintained at Neutral pH. AB - The formation of acetone and n-butanol by Clostridium acetobutylicum NCIB 8052 (ATCC 824) was monitored in batch culture at 35 degrees C in a glucose (2% [wt/vol]) minimal medium maintained throughout at either pH 5.0 or 7.0. At pH 5, good solvent production was obtained in the unsupplemented medium, although addition of acetate plus butyrate (10 mM each) caused solvent production to be initiated at a lower biomass concentration. At pH 7, although a purely acidogenic fermentation was maintained in the unsupplemented medium, low concentrations of acetone and n-butanol were produced when the glucose content of the medium was increased (to 4% [wt/vol]). Substantial solvent concentrations were, however, obtained at pH 7 in the 2% glucose medium supplemented with high concentrations of acetate plus butyrate (100 mM each, supplied as their potassium salts). Thus, C. acetobutylicum NCIB 8052, like C. beijerinckii VPI 13436, is able to produce solvents at neutral pH, although good yields are obtained only when adequately high concentrations of acetate and butyrate are supplied. Supplementation of the glucose minimal medium with propionate (20 mM) at pH 5 led to the production of some n-propanol as well as acetone and n-butanol; the final culture medium was virtually acid free. At pH 7, supplementation with propionate (150 mM) again led to the formation of n-propanol but also provoked production of some acetone and n butanol, although in considerably smaller amounts than were obtained when the same basal medium had been fortified with acetate and butyrate at pH 7. PMID- 16346679 TI - Measurement and study of substrate specificity of exoglucosidase activity in eutrophic water. AB - The alpha- and beta-glucosidase activity in natural samples can be readily measured during short incubation times (20 min) by using the artificial substrates 4-methylumbelliferyl-alpha-d-glucoside and 4-methylumbelliferyl-beta-d glucoside. The apparent K(m) of both alpha- and beta-glucosidase for these respective substrates is 0.01 muM. The homologous disaccharides maltose and cellobiose competitively inhibit alpha- and beta-glucosidase, respectively. Absolute substrate specificity of the alpha- and beta-glucosidase is observed with respect to the configuration of carbon atoms 1 and 4. Enrichment cultures on either alpha- and beta-glucoside result in increasing activity of the corresponding glucosidase, both in absolute terms and with respect to the other glucosidase. PMID- 16346680 TI - Root hair deformation, bacterial attachment, and plant growth in wheat azospirillum associations. AB - Seven Azospirillum strains induced more deformation of root hairs of wheat than did strains of Rhizobium leguminosarum, Azotobacter chroococcum, or Escherichia coli. Azospirillum sp. strain Sp245 caused the most deformation. Strain Sp245 (isolated from surface sterile roots of wheat) and strain Sp7 (isolated from the rhizosphere of a forage grass) were compared with regard to their effects on root hair deformation, their attachment to roots, and their effects on the growth of four wheat cultivars. The amount of deformation caused by the two strains in the four cultivars increased in the following order: cv. Tobari, cv. Tonari, cv. BH1146, cv. Lagoa. Strain Sp245 attached to the roots of all cultivars in low numbers, and attachment did not increase with time (up to 48 h). Strain Sp7 attached in higher numbers, and attachment increased with time. Inoculation of the four cultivars of wheat had pronounced effects on root mass measured at maturity. The magnitude of the effects in the four cultivars increased in the following order: Tobari, Tonari, BH1146, Lagoa; these effects were progressively more positive for strain Sp245 and progressively more negative for strain Sp7. Concentrations of N in wheat did not vary substantially between cultivars or strains. Concentrations of K and P did not vary substantially between cultivars but did vary between strains, Sp245 effecting increases and Sp7 effecting decreases. PMID- 16346681 TI - Assessing phytoplankton and bacterioplankton production during early spring in lake erken, sweden. AB - The spring development of both phytoplankton and bacterioplankton was investigated between 18 April and 7 May 1983 in mesotrophic Lake Erken, Sweden. By using the lake as a batch culture, our aim was to estimate, via different methods, the production of phytoplankton and bacterioplankton in the lake and to compare these production estimates with the actual increase in phytoplankton and bacterioplankton biomass. The average water temperature was 3.5 degrees C. Of the phytoplankton biomass, >90% was the diatom Stephanodiscus hantzchii var. pusillus, by the peak of the bloom. The C and O(2) methods of estimating primary production gave equivalent results (r = 0.999) with a photosynthetic quotient of 1.63. The theoretical photosynthetic quotient predicted from the C/NO(3) N assimilation ratio was 1.57. The total integrated incorporation of [C]bicarbonate into particulate material (>1 mum) was similar to the increase in phytoplankton carbon determined from cell counts. Bacterioplankton increased from 0.5 x 10 to 1.52 x 10 cells liter ( approximately 0.5 mug of C liter day). Estimates of bacterioplankton production from rates of [H]thymidine incorporation were ca. 1.2 to 1.7 mug of C liter day. Bacterial respiration, measured by a high-precision Winkler technique, was estimated as 4.8 mug of C liter day, indicating a bacterial growth yield of 25%. The bulk of the bacterioplankton production was accounted for by algal extracellular products. Gross bacterioplankton production (production plus respiration) was 20% of gross primary production, per square meter of surface area. We found no indication that bacterioplankton production was underestimated by the [H]thymidine incorporation method. PMID- 16346682 TI - Variation in Preference for Rhizobium meliloti Within and Between Medicago sativa Cultivars Grown in Soil. AB - Variation in nodulation preferences for Rhizobium strains within and between Medicago sativa cultivars was assessed in the greenhouse with plants grown in Leonard jars and two soils of diverse origin (Lanark and Ottawa), using inocula consisting of effective individual or paired strains of R. meliloti which could be recognized by high-concentration antibiotic resistance. The results indicated considerable variability in host preferences for R. meliloti among plants within cultivars but not between cultivars. The implications of this variation are discussed from the point of view of possible improvement of symbiotic nitrogen fixation. With one exception, the differences in nodulation success between inoculant R. meliloti strains were consistent in Leonard jars and both soils. All introduced strains formed significantly more nodules in Renfrew soil containing few native rhizobia than in Ottawa soil with a large resident R. meliloti population. Plants grown in Lanark soil without inoculation were ineffectively nodulated by native rhizobia and yielded significantly less growth than those receiving inoculation. In contrast, the yield of inoculated plants in Ottawa soil did not significantly differ from those without inoculation due to effective nodulation by native R. meliloti. The data indicated synergistic effects on yield by certain paired strain inocula relative to the same strains inoculated individually in Lanark but not in Ottawa soil or Leonard jars. PMID- 16346683 TI - Isolate 761M: a New Type I Methanotroph That Possesses a Complete Tricarboxylic Acid Cycle. AB - A methanotrophic bacterium, isolate 761M, grows slowly with methane as the sole carbon and energy source. Growth was stimulated by peptone, casein hydrolysate, glucose, and acetate plus malate. Sugars other than glucose did not stimulate growth. Growth yields, based on the amount of methane consumed, increased when other carbon sources were present, and less methane carbon was assimilated under these conditions. Methane was obligately required for growth of isolate 761M. This bacterium does not grow on rich media. Isolate 761M was found to possess hexulose phosphate synthase and intracytoplasmic membranes characteristic of other type I methanotrophs. Unlike other type I methanotrophs, this bacterium possessed alpha-ketoglutarate dehydrogenase and oxidized [2-C]acetate to carbon dioxide. PMID- 16346684 TI - Substrate degradation and pressure tolerance of free-living and attached bacterial populations in the intestines of shallow-water fish. AB - Bacterial populations attached to intestinal linings of shallow-water fish were compared to those free in the lumen for response to hydrostatic pressure and ability to degrade a variety of substrates. Results suggested that, unlike reports on gut-associated deep-sea bacteria, the two shallow-water populations were not significantly different in their pressure or substrate responsiveness. PMID- 16346685 TI - Protoplast Regeneration in Clostridium tertium: Isolation of Derivatives with High-Frequency Regeneration. AB - A procedure was developed for the formation and regeneration of protoplasts in Clostridium tertium. Although in the parent strain most of the protoplasts gave rise to L-form colonies, derivatives could be isolated which exhibited high regeneration frequencies. PMID- 16346686 TI - Underexpression of ap from R-plasmids in fast-growing Rhizobium species. AB - The presence of the plasmid RP1 in the cells of Rhizobium leguminosarum strains Rld1, 300, and 248, R. phaseoli 1233, R. trifolii strains T1 and 6661, and R. meliloti 4013 was found to appreciably increase bacterial resistance toward kanamycin and tetracycline but not toward ampicillin. The presence of 16 other R plasmids in R. leguminosarum was also found to either not increase or only marginally increase bacterial resistance toward ampicillin. It appears now that underexpression of the plasmid-specified ampicillin function is common to most fast- and slow-growing rhizobia. PMID- 16346687 TI - N(2)O evolution by green algae. AB - Evidence is presented here that axenic cultures of Chlorella, Scenedesmus, Coelastrum, and Chlorococcum spp. evolve N(2)O when grown on NO(2), showing that the Chlorophyceae are a source of N(2)O in aquatic systems. PMID- 16346688 TI - Effect of Culture Conditions on the Production of Tyrosine Phenol-Lyase by Erwinia herbicola. AB - The effect of environmental parameters on the growth and the tyrosine phenol lyase content of Erwinia herbicola was investigated. On mineral medium containing glycerol, l-tyrosine increased the enzyme content 23-fold. When the l-tyrosine was also the carbon source, bacterial growth was 300 times greater than the basal level. On a rich medium, tyrosine phenol-lyase production was strongly dependent on pH and aeration. Catabolite repression and induction both probably control enzyme content. PMID- 16346689 TI - Hydroxyproline-Rich Protein Material in Wood and Lignin of Fagus sylvatica. AB - The nitrogen content, distribution, and amino acid composition of protein material were determined in wood and lignin of Fagus sylvatica. The data indicated that the nitrogen originated from hydroxyproline-rich cell wall glycoprotein, about half of which may be bound to the lignin polymer. The implications for lignocellulose biodegradation are discussed. PMID- 16346690 TI - Role of chemotaxis in the ecology of denitrifiers. AB - A modification of the Adler capillary assay was used to evaluate the chemotactic responses of several denitrifiers to nitrate and nitrite. Strong positive chemotaxis was observed to NO(3) and NO(2) by soil isolates of Pseudomonas aeruginosa, Pseudomonas fluorescens, and Pseudomonas stutzeri, with the peak response occurring at 10 M for both attractants. In addition, a strong chemoattraction to serine (peak response at 10 M), tryptone, and a soil extract, but not to NH(4), was observed for all denitrifiers tested. Chemotaxis was not dependent on a previous growth on NO(3), NO(2), or a soil extract, and the chemoattraction to NO(3) occurred when the bacteria were grown aerobically or anaerobically. However, the best response to NO(3) was usually observed when the cells were grown aerobically with 10 mM NO(3) in the growth medium. Capillary tubes containing 103 M NO(3) submerged into soil-water mixtures elicited a significant chemotactic response to NO(3) by the indigenous soil microflora, the majority of which were Pseudomonas spp. A chemotactic strain of P. fluorescens also was shown to survive significantly better in aerobic and anaerobic soils than was a nonmotile strain of the same species. Both strains had equal growth rates in liquid cultures. Thus, chemotaxis may be one mechanism by which denitrifiers successfully compete for available NO(3) and NO(2), and which may facilitate the survival of naturally occurring populations of some denitrifiers. PMID- 16346691 TI - Enrichment and Isolation of Rumen Bacteria That Reduce trans- Aconitic Acid to Tricarballylic Acid. AB - Bacteria from the bovine rumen capable of reducing trans-aconitate to tricarballylate were enriched in an anaerobic chemostat containing rumen fluid medium and aconitate. After 9 days at a dilution rate of 0.07 h, the medium was diluted and plated in an anaerobic glove box. Three types of isolates were obtained from the plates (a crescent-shaped organism, a pleomorphic rod, and a spiral-shaped organism), and all three produced tricarballylate in batch cultures that contained glucose and trans-aconitate. In glucose-limited chemostats (0.10 h), trans-aconitate reduction was associated with a decrease in the amount of reduced products formed from glucose. The crescent-shaped organism produced less propionate, the pleomorphic rod produced less ethanol, and the spiral made less succinate and possibly H(2). Aconitate reduction by the pleomorphic rod and the spiral organism was associated with a significant increase in cellular dry matter. Experiments with stock cultures of predominant rumen bacteria indicated that Selenomonas ruminantium, a species taxonomically similar to the crescent shaped isolate, was an active reducer of trans-aconitate. Strains of Bacteroides ruminicola, Butyrivibrio fibrisolvens, and Megasphaera elsdenii produced little if any tricarballylate. Wolinella succinogenes produced some tricarballylate. Based on its stability constant for magnesium (K(eq) = 115), tricarballylate could be a factor in the hypomagnesemia that leads to grass tetany. PMID- 16346692 TI - Effectiveness of Rhizobium strains used in inoculants after their introduction into soil. AB - Rhizobium strains used in inoculants for Trifolium spp., Medicago spp., Glycine max, and Lotus pedunculatus were isolated from nodules of these legumes grown in soils into which the rhizobia had been introduced 4 to 8 years before. Isolations were made from a total of 420 nodules. Nodule occupancy by the inoculant strains varied from 17.7% for a soybean strain to 100% in the case of L. pedunculatus whose specific rhizobia did not occur in the soils studied. In general, inoculant strains isolated from nodules did not differ in effectiveness from cultures of the same strains concurrently maintained in lyophilized form. The average effectiveness of all of the isolates (identified and unidentified) from a legume was 7.1 to 73.3% higher than that of the unidentified isolates alone, demonstrating the prolonged effect that a single-seed inoculation has on the rhizobial population in a soil which had not been planted with legumes before. Relatively weak recovery of a Rhizobium japonicum strain introduced into soil 4 years after soybean seed inoculated with a different strain had been planted in the same soil confirmed the advantage of a resident population over an introduced inoculant strain. PMID- 16346693 TI - Degassing of Pore Water Methane during Sediment Incubations. AB - Laboratory experiments were used to examine the degassing of CH(4) from a muddy sediment. Sediment containing dissolved CH(4) showed a hyperbolic time course of CH(4) release when allowed to degas in stoppered 20-ml vials. Equilibration required ca. 24 h for 5 ml of sediment. The rate of CH(4) release was found to be dependent on the ratio of exposed sediment surface area to sediment volume. The water content of the sediment was a factor in the total amount of CH(4) released but did not affect the rate of degassing. Addition of water to sediment samples (to form a slurry) accelerated CH(4) release, with a 1:1 dilution giving ca. 80% of maximum release after only 2 min. Shaking (vortexing) the sediments also facilitated CH(4) exchange, with 2 min of vigorous agitation giving 77% of maximum release. The organic content of the sediment did not affect either the amount or the rate of CH(4) degassing. Rubber stoppers exposed to CH(4) were found to absorb CH(4) rapidly and to subsequently release it in proportion to the concentration to which they were exposed. Artifacts may be associated with CH(4) production measurements if sediment and stopper degassing are not considered. It is recommended that any study of methane production or distribution include preliminary experiments to determine the degassing kinetics for the specific sediment system being used. PMID- 16346694 TI - d-Glucose Transport System of Zymomonas mobilis. AB - The properties of the d-glucose transport system of Zymomonas mobilis were determined by measuring the uptake of nonmetabolizable analogs (2-deoxy-d-glucose and d-xylose) by wild-type cells and the uptake of d-glucose itself by a mutant lacking glucokinase. d-Glucose was transported by a constitutive, stereospecific, carrier-mediated facilitated diffusion system, whereby its intracellular concentration quickly reached a plateau close to but not above the external concentration. d-Xylose was transported by the d-glucose system, as evidenced by inhibition of its uptake by d-glucose. d-Fructose was not an efficient competitive inhibitor of d-glucose uptake, indicating that it has a low affinity for the d-glucose transport system. The apparent K(m) of d-glucose transport was in the range of 5 to 15 mM, with a V(max) of 200 to 300 nmol min mg of protein. The K(m) of Z. mobilis glucokinase (0.25 to 0.4 mM) was 1 order of magnitude lower than the K(m) for d-glucose transport, although the V(max) values for transport and phosphorylation were similar. Thus, glucose transport cannot be expected to be rate limiting at concentrations of extracellular glucose normally used in fermentation processes, which greatly exceed the K(m) for the transport system. The low-affinity, high-velocity, nonconcentrative system for d-glucose transport described here is consistent with the natural occurrence of Z. mobilis in high-sugar environments and with the capacity of Z. mobilis for rapid conversion of glucose to metabolic products with low energetic yield. PMID- 16346695 TI - Influence of Serratia marcescens Pigmentation on Cell Concentrations in Aerosols Produced by Bursting Bubbles. AB - For eight strains of Serratia marcescens, increased cell concentrations were found in aerosols produced from bursting bubbles, with concentrations ranging from a maximum of ca. 80 times the bulk concentration for pigmented strains 4180, 933, and 274 to a minimum approximately equal to the bulk concentration for nonpigmented strain 8100. The increased cell concentration in the aerosol was suppressed when pigmented strains were grown at 37 degrees C, a temperature at which the pigment prodigiosin is not synthesized, resulting in lower concentrations similar to those of nonpigmented strains. Strains that produce higher concentrations of prodigiosin after 1, 2, 4, and 8 days of growth show increasing concentrations in bubble-produced drops; duplicate cultures grown at 37 degrees C did not show any increases. In four concurrent experiments, cells starved for 24 h showed greater concentrations than nonstarved cells for chromogenic strain NIMA, whereas for nonchromogenic strain WF, starved cells showed greater concentrations in three cases and a decreased concentration in the fourth. Bacterial concentrations in aerosol drops from bursting bubbles appear to be predominantly influenced by the surface condition of the bacterial cell. PMID- 16346696 TI - Sulfate reduction in freshwater sediments receiving Acid mine drainage. AB - One arm of Lake Anna, Va., receives acid mine drainage (AMD) from Contrary Creek (SO(4) concentration = 2 to 20 mM, pH = 2.5 to 3.5). Acid-volatile sulfide concentrations, SO(4) reduction rates, and interstitial SO(4) concentrations were measured at various depths in the sediment at four stations in four seasons to assess the effects of the AMD-added SO(4) on bacterial SO(4) reduction. Acid volatile sulfide concentrations were always an order of magnitude higher at the stations receiving AMD than at a control station in another arm of the lake that received no AMD. Summer SO(4) reduction rates were also an order of magnitude higher at stations that received AMD than at the control station (226 versus 13.5 mmol m day), but winter values were inconclusive, probably due to low sediment temperature (6 degrees C). Profiles of interstitial SO(4) concentrations at the AMD stations showed a rapid decrease with depth (from 1,270 to 6 muM in the top 6 cm) due to rapid SO(4) reduction. Bottom-water SO(4) concentrations in the AMD receiving arm were highest in winter and lowest in summer. These data support the conclusion that there is a significant enhancement of SO(4) reduction in sediments receiving high SO(4) inputs from AMD. PMID- 16346697 TI - Solid-State Fermentation with Trichoderma reesei for Cellulase Production. AB - Cellulase yields of 250 to 430 IU/g of cellulose were recorded in a new approach to solid-state fermentation of wheat straw with Trichoderma reesei QMY-1. This is an increase of ca. 72% compared with the yields (160 to 250 IU/g of cellulose) in liquid-state fermentation reported in the literature. High cellulase activity (16 to 17 IU/ml) per unit volume of enzyme broth and high yields of cellulases were attributed to the growth of T. reesei on a hemicellulose fraction during its first phase and then on a cellulose fraction of wheat straw during its later phase for cellulase production, as well as to the close contact of hyphae with the substrate in solid-state fermentation. The cellulase system obtained by the solid-state fermentation of wheat straw contained cellulases (17.2 IU/ml), beta glucosidase (21.2 IU/ml), and xylanases (540 IU/ml). This cellulase system was capable of hydrolyzing 78 to 90% of delignified wheat straw (10% concentration) in 96 h, without the addition of complementary enzymes, beta-glucosidase, and xylanases. PMID- 16346698 TI - Degradation of dehydrodivanillin by anaerobic bacteria from cow rumen fluid. AB - Dehydrodivanillin (DDV; 0.15 g/liter) was biodegradable at 37 degrees C under strictly anaerobic conditions by microflora from cow rumen fluid to the extent of 25% within 2 days in a yeast extract medium. The anaerobes were acclimated on DDV for 2 weeks, leading to DDV-degrading microflora with rates of degradation eight times higher than those initially. Dehydrodivanillic acid and vanillic acid were detected in an ethylacetate extract of a DDV-enriched culture broth by thin layer, gas, and high-performance liquid chromatographies and by mass spectrometry. PMID- 16346699 TI - Sulfide alleviation of the acetylene inhibition of nitrous oxide reduction in soil. AB - The alleviation of the acetylene blockage of nitrous oxide reduction by sulfide was studied in anaerobically incubated Brookston soil to better characterize the process. Removal of nitrate-derived nitrous oxide from soil amended with acetylene and sulfide occurred earlier in the presence of glucose than it did in its absence. This was attributed to the influence of glucose on nitrous oxide production rather than reduction during the early stages of the soil incubation. Glucose was found to have no effect on reduction of injected nitrous oxide in the presence of acetylene- and sulfide-amended soil, whereas carbon dioxide significantly stimulated reduction. It is suggested that the microorganism(s) involved may use carbon dioxide as a cellular carbon source. The sulfide added to the soil probably did not act solely as an electron donor, as the number of electrons required to reduce the added nitrous oxide in our systems was greater than the amount supplied by the sulfide. The soil pH at which the alleviation occurred was 6.7 and was not affected by the sulfide treatment. PMID- 16346700 TI - Autoradiographic Studies of [methyl-H]Thymidine Incorporation in a Cyanobacterium (Microcystis wesenbergii)-Bacterium Association and in Selected Algae and Bacteria. AB - The present investigation showed by means of autoradiography that the cyanobacterium Microcystis wesenbergii did not incorporate [H]thymidine at nanomolar concentrations, whereas its associated heterotrophic bacteria appearing in the gelatinous cover of the cyanobacterium became labeled. Several other tested cyaobacteria and algae did not incorporate [H]thymidine. PMID- 16346701 TI - Response of bacteria isolated from a pristine prairie stream to concentration and source of soluble organic carbon. AB - Responses of native aquatic bacteria to source and concentration of dissolved organic carbon was observed by using gradient plates. Dissolved organic carbon of prairie (grasses) or gallery forest (bur oak) origin was used as the carbon source in these observations. Bacteria isolated from sediments in the grassland reaches of a prairie stream showed growth only on grass leachates. Bacteria isolated from the gallery forest reaches of the same stream were able to grow on plates made from either source of leachate. The differing quality, quantity, and rate of supply of these dissolved organic carbon sources should select for variation in the bacterial assemblages. PMID- 16346702 TI - Hybridization and Polyploidization of Saccharomyces cerevisiae Strains by Transformation-Associated Cell Fusion. AB - Hybrid or polyploid clones of Saccharomyces cerevisiae produced by protoplast fusion were easily isolated by selecting transformants with the plasmid phenotype because the transformation was directly associated with cell fusion. When haploid cells were used as the original strain, the transformants were mostly diploids with a significant fraction of polyploids (triploids or tetraploids). Repeated transformation after curing the plasmid gave rise to clones with higher ploidy, but the frequency of cell fusion was severely reduced as ploidy increased. PMID- 16346703 TI - Production and Consumption of H(2) during Growth of Methanosarcina spp. on Acetate. AB - Methanosarcina sp. strain TM-1 and Methanosarcina acetivorans produced and consumed H(2) to maintain H(2) partial pressures of 16 to 92 Pa in closed cultures during growth on acetate. Strain TM-1 produced H(2) continuously when H(2) was continuously removed from the culture. The potential physiological significance of H(2) in acetate metabolism to methane is discussed. PMID- 16346704 TI - Cytochemical Localization of Urease in a Rumen Staphylococcus sp. by Electron Microscopy. AB - We describe a technique whereby intracellular urease activity can be localized by transmission electron microscopy. The ammonia produced from the enzymatic hydrolysis of urea is first precipitated with sodium tetraphenylboron and then replaced with silver to produce electron-dense silver tetraphenylboron. This direct reaction product deposition procedure was used to demonstrate the presence of membrane-bound urease of Staphylococcus sp. H3-22, a gram-positive ruminal bacterium. PMID- 16346705 TI - New selective media for enumeration and recovery of fluorescent pseudomonads from various habitats. AB - New media (S1 and S2) were formulated that provide a high degree of selectivity and detection of fluorescent pseudomonads on initial plating. The selectivity of the S-type media was based on a detergent, sodium lauroyl sarcosine, and an antibiotic, trimethoprim. A total of five soils from different geographical locations and one sewage sludge sample were examined. On S1 medium, isolates from two soils with low fluorescent pseudomonad populations exhibited a high frequency of arginine dihydrolase (78%) and oxidase-positive (95%) phenotypes, but no fermentative isolates were recovered. Medium S2 was more defined and selective than S1, but lower numbers of fluorescent pseudomonads were recovered on S2. In soils in which fluorescent pseudomonads represent a small proportion of the total population, S1 medium consistently recovered high percentages of fluorescent phenotypes (82.5%). PMID- 16346706 TI - Change in Microbial Numbers during Thermophilic Composting of Sewage Sludge with Reference to CO(2) Evolution Rate. AB - Dewatered sewage sludge was composted in a laboratory-scale autothermal reactor in which a constant temperature of 60 degrees C was kept as long as possible by regulating the air feed rate. The change in CO(2) evolution rate was measured continuously from the start up through the cessation of compositing. The succession of mesophilic bacteria, thermophilic bacteria, and thermophilic actinomycetes was also observed during the composting. Specific CO(2) evolution rates of thermophilic bacteria and actinomycetes in the constant-temperature region of 60 degrees C were assessed quantitatively. It was found that the CO(2) evolution rate was attributed to thermophilic bacteria at the initial stage of 60 degrees C and to thermophilic actinomycetes at the later stage of 60 degrees C. PMID- 16346707 TI - Characteristics of Mesophilic Bacteria Isolated during Thermophilic Composting of Sewage Sludge. AB - The degree of inactivation by UV irradiation was different between vegetative cells and spores of bacteria isolated from sewage sludge composting at 60 degrees C. By using this property, a method to estimate the spore ratio of a mixture of vegetative cells and spores was presented. This UV irradiation method was applied to the estimation of the spore ratio of sewage sludge compost samples collected at several stages of composting. The spore ratio of mesophilic bacteria in the samples obtained at the thermophilic stage of 60 degrees C was 40% at most. The vegetative form of mesophilic bacteria showed a thermotolerance property at 60 degrees C by forming colonies but showed no respiratory activity at that temperature. PMID- 16346708 TI - Effects of Some Environmental Factors on Growth Characteristics of Candida utilis on Peat Hydrolysates. AB - Samples of peat from Pine Island and Brookston bogs in Minnesota were hydrolyzed with various concentrations of HCl or H(2)SO(4) solutions, before or after debituminization (an extraction process used to remove waxy materials, bitumens, from peat), to produce peak hydrolysates as growth substrates for Candida utilis. Hydrolysates were neutralized with concentrated NaOH solution to pH 3.5, 4.5, 5.0, 5.5, 6.0, and 7.0. The precipitated humates were removed by filtration. The resulting peat hydrolysates were amended with reagent-grade K(2)HPO(4), K(2)SO(4), and MgSO(4), 200, 100, and 50 mg per liter of peat hydrolysate, respectively. The debituminized peat produced more total nitrogen (TN) and reducing substances (RS) than the nondebituminized peat. Peat hydrolysates produced by HCl solutions contained slightly higher RS and TN than those produced by H(2)SO(4) solutions; however, the latter were better growth substrates than the former. The yield coefficients in both H(2)SO(4) and HCl hydrolysates initially decreased at 12 to 24 h and then increased gradually over the remaining incubation period (24 to 96 h). As TN and RS were increased, an increase in cell density, biomass, and productivity was observed. In contrast, a decrease in specific growth rate occurred as the RS and TN were increased. The generation time of C. utilis was affected by the concentrations of RS and TN. A peak substrate yield coefficient was found at pH 5.0 in HCl hydrolysates and at pH 6.0 to 6.5 in H(2)SO(4) hydrolysates. Good linear correlation coefficients were found between RS and biomass of C. utilis. The coefficients of correlation increased as the TN level in hydrolysates was increased. PMID- 16346709 TI - Root Hair Deformations Associated with Fractionated Extracts from Rhizobium trifolii. AB - Components from culture fluid and whole cells of Rhizobium trifolii were examined for effects on root hair morphology of white clover seedlings (Trifolium repens var. Ladino). Cell-free culture fluid, exopolysaccharides, supernatant fluid from the precipitation of the exopolysaccharides, capsular polysaccharides, lipopolysaccharides, and a protein fraction from culture fluids were assayed for morphogenetic effects on the root hairs of axenically grown clover seedlings. Crude fractions were chromatographed on Bio Gel A-5m (Bio-Rad Laboratories), and fractions collected were similarly assayed. Hexose, uronic acid, and protein concentrations were determined for all fractions assayed. Gel chromatography indicated the materials with deforming ability to be of high molecular weight (>10,000). For all fractions except exopolysaccharide, deforming ability was associated with a protein component. This suggested that two components were associated with deformation; both contained polysaccharides and one contained protein. Crude fractions differed in their ability to cause deformations and indicated the following relative ability (in decreasing order) to deform root hairs: cell-free culture fluid, capsular polysaccharides, protein from culture fluids, exopolysaccharide, and cell envelope. Lipopolysaccharides had no effect. PMID- 16346710 TI - Factors Affecting the Germination of Akinetes of Nodularia spumigena (Cyanobacteriaceae). AB - Nutritional and physical factors which influence the germination of akinetes of Nodularia spumigena (Cyanobacteriaceae) were examined. Low concentrations of phosphorus (<0.9 muM) were required for germination. Nitrate had no effect, but ammonia, at concentrations of >45 muM, inhibited germination. Salinities of >20 per thousand were inhibitory to germination. Optimum temperatures were 22 degrees C or greater. Germination did not take place in the dark, but only very low light intensities (0.5 microeinstein m s) were necessary to initiate germination. Red light (620 to 665 nm) was required. More than 24 h of continuous exposure to light was necessary for any significant germination to occur. The conditions for germination corresponded with conditions in the Peel-Harvey Estuary, Western Australia, 2 to 3 weeks before large summer Nodularia blooms. PMID- 16346711 TI - Localization of Polyvinyl Alcohol Oxidase Produced by a Bacterial Symbiont, Pseudomonas sp. Strain VM15C. AB - An axenic culture of a polyvinyl alcohol (PVA)-degrading symbiont, Pseudomonas sp. strain VM15C, was established on PVA with a crude preparation of the growth factor (factor A) produced by the symbiotic partner Pseudomonas putida VM15A. An increase of factor A in the culture medium enhanced the cell-associated PVA oxidase activity as well as the growth rate, but decreased production of extracellular PVA oxidase. PVA oxidase in cells grown on PVA was present in the periplasmic space at a higher ratio than in cells grown on peptone. PVA degradation occurred rapidly with washed cells. PVA was also degraded by immobilized cells entrapped in agar gels. PMID- 16346712 TI - Characterization of a Marine Bacterium Associated with Crassostrea virginica (the Eastern Oyster). AB - A gram-negative bacterium found to be closely associated with oysters has been isolated and characterized. The organism, designated LST, has a generation time of 106 min in Marine broth under optimal growth conditions at 25 degrees C. During the decline phase of growth, it exhibits a morphological transition from a motile rod (ca. 1 mum in length) to an elongated, 3- to 40-mum, nonmotile, tightly coiled helix. LST synthesizes and releases a pigment in the stationary and decline phases of growth. Identified as melanin on the basis of chemical properties and UV absorbance maxima, the pigment comprises polymers of heterogeneous molecular weights, ranging from 12,000 to 120,000. The guanosine plus-cytosine content of the LST DNA is 46%, and results of phenetic analysis and DNA-DNA hybridization indicate that this bacterium represents a new species. LST adheres to a variety of surfaces, including glass, plastics, and oyster shell, and has been shown to promote the settlement of oyster larvae. PMID- 16346713 TI - Control of Oxidative Sulfur Metabolism of Chlorobium limicola forma thiosulfatophilum. AB - A metered blend of anaerobic-grade N(2), CO(2), and H(2)S gases was introduced into an illuminated, 800-ml liquid volume, continuously stirred tank reactor. The system, described as an anaerobic gas-to-liquid phase fed-batch reactor, was used to investigate the effects of H(2)S flow rate and light energy on the accumulation of oxidized sulfur compounds formed by the photoautotroph Chlorobium limicola forma thiosulfatophilum during growth. Elemental sulfur was formed and accumulated in stoichiometric quantities when light energy and H(2)S molar flow rate levels were optimally adjusted in the presence of nonlimiting CO(2). Deviation from the optimal H(2)S and light energy levels resulted in either oxidation of sulfur or complete inhibition of sulfide oxidation. Based on these observations, a model of sulfide and sulfur oxidases electrochemically coupled to the photosynthetic reaction center of Chlorobium spp. is presented. The dynamic deregulation of oxidative pathways may be a mechanism for supplying the photosynthetic reaction center with a continuous source of electrons during periods of varying light and substrate availability, as in pond ecosystems where Chlorobium spp. are found. Possible applications for a sulfide gas removal process are discussed. PMID- 16346714 TI - Production and Characterization of Polymeric Lignin Degradation Intermediates from Two Different Streptomyces spp. AB - Previous investigations have identified a quantitatively major intermediate of lignin degradation by Streptomyces viridosporus. The intermediate, a modified lignin polymer, acid-precipitable polymeric lignin (APPL), is released as a water soluble catabolite and has been recovered in amounts equivalent to 30% of the lignin originally present in a corn stover lignocellulose substrate after degradation by this actinomycete. In the present work, APPLs were collected at various time intervals from cultures of two highly ligninolytic Streptomyces sp. strains, S. viridosporus T7A and S. badius 252, growing on corn stover lignocellulose. APPL production was measured over time, and the chemistry of APPLs produced by each organism after different time intervals was compared. Chemical characterizations included assays for lignin, carbohydrate, and ash contents, molecular weight distributions by gel permeation chromatography, and chemical degradation analyses by permanganate oxidation, acidolysis, and alkaline ester hydrolysis. Differences between the organisms were observed in the cultural conditions required for APPL production and in the time courses of APPL accumulation. S. viridosporus produced APPL in solid-state fermentation over a 6- to 8-week incubation period, whereas S. badius produced as much or more APPL, but only in liquid culture and over a 7- to 8-day incubation period. The chemistry of the APPLs produced also differed. S. viridosporus APPL was more lignin-like than that of S. badius and was slowly modified further over time, although no change in molecular weight distribution over time was observed. In contrast, S. badius APPL was less lignin-like and increased substantially in average molecular weight over time. Results indicated that differing mechanisms of lignin metabolism may exist in these two Streptomyces sp. strains. S. viridosporus APPL probably originates from the heart of the lignin and is released largely as the result of beta-ether cleavage and other oxidative reactions. S. badius APPL probably originates in the same manner; however, after release as a water-soluble catabolite, lower-molecular-weight intermediates of lignin degradation are repolymerized with APPL in a reaction catalyzed by an extracellular phenol oxidase. The chemical analyses and the presence of extracellular phenol oxidase in S. badius, but not in S. viridosporus, support this conclusion. PMID- 16346715 TI - Bacterial o-methylation of chloroguaiacols: effect of substrate concentration, cell density, and growth conditions. AB - O-methylation of chloroguaiacols has been examined in a number of gram-positive and gram-negative bacteria to elucidate the effects of substrate concentration, growth conditions, and cell density. Substrate concentrations between 0.1 and 20.0 mg liter were used, and it was found that (i) yields of the O-methylated products were significantly higher at the lowest concentrations and (ii) rates of O-methylation were not linear functions of concentration. With 3,4,5 trichloroguaiacol, the nature of the metabolites also changed with concentration. During growth with a range of substrates, O-methylation of chloroguaiacols also took place. With vanillate, however, de-O-methylation occurred: the chlorocatechol formed from 4,5,6-trichloroguaiacol was successively O-methylated to 3,4,5-trichloroguaiacol and 3,4,5-trichloroveratrole, whereas that produced from 4,5-dichloroguaiacol was degraded without O-methylation. Effective O methylation in nonproliferating suspensions occurred at cell densities as low as 10 cells ml, although both the yields and the rates were lower than in more dense cultures. By using disk assays, it was shown that, compared with their precursors, all of the O-methylated metabolites were virtually nontoxic to the strains examined. It is therefore proposed that O-methylation functions as a detoxification mechanism for cells exposed to chloroguaiacols and chlorophenols. In detail, significant differences were observed in the response of gram-positive and gram-negative cell strains to chloroguaiacols. It is concluded that bacterial O-methylation is to be expected in the natural environment subjected to discharge of chloroguaiacols. PMID- 16346716 TI - Factors Involved in the Regulation of a Ligninase Activity in Phanerochaete chrysosporium. AB - The regulation of an H(2)O(2)-dependent ligninolytic activity was examined in the wood decay fungus Phanerochaete chrysosporium. The ligninase appears in cultures upon limitation for nitrogen or carbohydrate and is suppressed by excess nutrients, by cycloheximide, or by culture agitation. Activity is increased by idiophasic exposure of cultures to 100% O(2). Elevated levels of ligninase and, in some cases, of extracellular H(2)O(2) production are detected after brief incubation of cultures with lignins or lignin substructure models, with the secondary metabolite veratryl alcohol, or with other related compounds. It is concluded that lignin degradation (lignin --> CO(2)) by this organism is regulated in part at the level of the ligninase, which is apparently inducible by its substrates or their degradation products. PMID- 16346717 TI - Persistence of denitrifying enzyme activity in dried soils. AB - The effects of air drying soil on denitrifying enzyme activity, denitrifier numbers, and rates of N gas loss from soil cores were measured. Only 29 and 16% of the initial denitrifying enzyme activity in fresh, near field capacity samples of Maury and Donerail soils, respectively, were lost after 7 days of air drying. The denitrifying activity of bacteria added to soil and activity recently formed in situ were not stable during drying. When dried and moist soil cores were irrigated, evolution of N gas began, and it maximized sooner in the dried cores. This suggests that the persistence of denitrifying enzymes permits accelerated denitrification when dried soils are remoistened. Enzyme activity increased significantly in these waterlogged cores, but fluctuations in enzyme activity were small compared with fluctuations in actual denitrification rate, and enzyme activities were always greater than denitrification rates. Apparent numbers of isolatable denitrifiers (most-probable-number counts) decreased more than enzyme activity as the soils were dried, but after the soils were rewetted, the extent of apparent growth was not consistently related to the magnitude of N loss. We hypothesize that activation-inactivation of existing enzymes by soil O(2) is of greater significance in transient denitrification events than is growth of denitrifiers or synthesis of new enzymes. PMID- 16346718 TI - Role of the Cell Surface of Methanosarcina mazei in Cell Aggregation. AB - Colonial aggregates of Methanosarcina (= Methanococcus) mazei were examined with scanning and transmission electron microscopy. Cells are irregular and grouped into multicellular sarcinal colonies, which may disaggregate in older cultures. The protoplast is bounded by a typical trilaminar plasma membrane, outside of which is a matrix of loose fibrils. The presence and compactness of matrix material are responsible for the close packing of cells, and colony disaggregation seems to be the result of matrix shedding and degradation. The cell envelope contains complex hetero polysaccharides of N-acetylgalactosamine and galacturonic and glucuronic acids. Polymers extruded by M. mazei are likely quite adhesive in nature, accounting for its strong adherence to surfaces and hardiness compared with many other methanogens. PMID- 16346719 TI - Partial Isolation and Degradation of Caseins by Cell Wall Proteinase(s) of Streptococcus cremoris HP. AB - The cell wall proteinase fraction of Streptococcus cremoris HP has been isolated. This preparation did not exhibit any activity due to either specific peptidases known to be located near the outside surface of and in the membrane or intracellular proteolytic enzymes. By using thin-layer chromatography for the detection of relatively small hydrolysis products which remain soluble at pH 4.6, it was shown that beta-casein is preferentially attacked by the cell wall proteinase. This was also the case when whole casein or micelles were used as the substrate. kappa-casein hydrolysis is a relatively slow process, and alpha(s) casein degradation appeared to proceed at an extremely low rate. These results could be confirmed by using CH(3)-labeled caseins. A relatively fast and linear initial progress of CH(3)-labeled beta-casein degradation is not inhibited by alpha(s)-casein and only slightly by kappa-casein at concentrations of these components which reflect their stoichiometry in the micelles. Possible implications of beta-casein degradation for growth of the organism in milk are discussed. PMID- 16346720 TI - Effect of metal-rich sewage sludge application on the bacterial communities of grasslands. AB - The effect of long-term application of heavy metal-laden sewage sludge on the total heterotrophic aerobic and the cadmium-resistant soil bacterial communities was studied. Gram-positive bacteria were completely absent from resistant communities. These findings suggest that this group is highly susceptible to Cd. Shannon's diversity indices estimated for total communities did not reveal negative effects on the communities that developed in the presence of sludge. However, Cd-resistant communities isolated from long-term sludge-amended soils were more diverse than the resistant communities from a control sample, suggesting that adaptation to Cd as a stressor had occurred in the presence of sludge constituents. This higher diversity was attributed to Cd resistance in pseudomonads and gram-negative fermenters. Resistance did not develop by dissemination of Cd resistance plasmids, because these were rarely detected in the genomes of resistant strains. PMID- 16346721 TI - Anaerobic degradation of soluble fractions of [C-lignin]lignocellulose. AB - [C-lignin]lignocellulose was solubilized by alkaline heat treatment and separated into different molecular size fractions for use as the sole source of carbon in anaerobic enrichment cultures. This study is aimed at determining the fate of low molecular-weight, polyaromatic lignin derivatives during anaerobic degradation. Gel permeation chromatography was used to preparatively separate the original C lignin substrate into three component molecular size fractions, each of which was then fed to separate enrichment cultures. Biodegradability was assessed by monitoring total carbon dioxide and methane production, evolution of labeled gases, loss of C-activity from solution, and changes in gel permeation chromatographic elution patterns. Results indicated that the smaller the size of the molecular weight fraction, the more extensive the degradation to gaseous end products. In addition, up to 30% of the entire soluble lignin-derived carbon was anaerobically mineralized to carbon dioxide and methane. PMID- 16346722 TI - Aromatic and Volatile Acid Intermediates Observed during Anaerobic Metabolism of Lignin-Derived Oligomers. AB - Anaerobic enrichment cultures acclimated for 2 years to use a C-labeled, lignin derived substrate with a molecular weight of 600 as a sole source of carbon were characterized by capillary and packed column gas chromatography. After acclimation, several of the active methanogenic consortia were inhibited with 2 bromoethanesulfonic acid, which suppressed methane formation and enhanced accumulation of a series of metabolic intermediates. Volatile fatty acids levels in 2-bromoethanesulfonic acid-amended cultures were 10 times greater than those in the uninhibited, methane-forming consortia with acetate as the predominant component. Furthermore, in the 2-bromoethanesulfonic acid-amended consortia, almost half of the original substrate carbon was metabolized to 10 monoaromatic compounds, with the most appreciable quantities accumulated as cinnamic, benzoic, caffeic, vanillic, and ferulic acids. 2-Bromoethanesulfonic acid seemed to effectively block CH(4) formation in the anaerobic food chain, resulting in the observed buildup of volatile fatty acids and monoaromatic intermediates. Neither fatty acids nor aromatic compounds were detected in the oligolignol substrate before its metabolism, suggesting that these anaerobic consortia have the ability to mediate the cleavage of the beta-aryl-ether bond, the most common intermonomeric linkage in lignin, with the subsequent release of the observed constituent aromatic monomers. PMID- 16346723 TI - Flocculation effects on bound water in sludges as measured by nuclear magnetic resonance spectroscopy. AB - Nuclear magnetic resonance relaxation times (T(1) and T(2)) were measured for flocculated and unflocculated samples of activated sludge. The weight of water and solids in the sludge samples was found and related to T(1) to find the relative percentage of bound water. The results suggest that the amount of bound water increases as the samples become more unflocculated. The values of T(1) and T(2) also indicate that unflocculated individual particles are characterized by loose packing of shorter molecules and that the addition of larger molecules may induce flocculation. PMID- 16346724 TI - Solid-state fermentation: an alternative to improve the nutritive value of coffee pulp. AB - Coffee pulp was subjected to a solid-state fermentation process, using Aspergillus niger. The initial moisture content of the pulp, as well as the fermentation time and temperature, had a significant effect on the increase in total amino acid content of the material. The increase in total amino acids showed a significant correlation with the dry matter recovered (r = -0.98) and the increase in pH during the process (r = 0.98). With a moisture content of 80%, a pH of 3.5, a temperature of 35 degrees C, and an aeration of 8 liters/min per kg as fermentation conditions, it was found that the maximum concentration of total amino acids was attained after 43 h. The fermented product had a higher total amino acid content and a lower cell wall constituent value (primarily cellulose and hemicellulose) than the original pulp. A growing chicken's ration containing 10% of the fermented product had a feed efficiency (2.14) similar to that of the standard ration (2.19) and was significantly better than that of the diet containing 10% of the original pulp (2.53). The difference observed in feed intake and weight gain between the standard diet and that with 10% of the fermented product is considered to be due to palatability factors which should be studied further. PMID- 16346725 TI - Influence of easily degradable naturally occurring carbon substrates on biodegradation of monosubstituted phenols by aquatic bacteria. AB - The influence of readily degradable, naturally occurring carbon substrates on the biodegradation of several monosubstitued phenols (m-cresol, m-aminophenol, p chlorophenol) was examined. The natural substrate classes used were amino acids, carbohydrates, and fatty acids. Samples of the microbial community from Lake Michie, a mesotrophic reservoir, were adapted to different levels of representatives from each natural substrate class in chemostats. After an extended adaptation period, the ability of the microbial community to degrade the monosubstituted phenols was determined by using a radiolabeled substrate uptake and mineralization method. Several microbiological characteristics of the communities were also measured. Adaptation to increasing concentrations of amino acids, carbohydrates, or fatty acids enhanced the ability of the microbial community to degrade all three phenols. The stimulation was largest for m-cresol and m-aminophenol. The mechanism responsible for the enhancement of monosubstituted phenol metabolism was not clearly identified, but the observation that adaptation to amino acids also increased the biodegradation of glucose and, to a lesser extent, naphthalene suggests a general stimulation of microbial metabolism. This study demonstrates that prior exposure to labile, natural substrates can significantly enhance the ability of aquatic microbial communities to respond to xenobiotics. PMID- 16346726 TI - Influence of naturally occurring humic acids on biodegradation of monosubstituted phenols by aquatic bacteria. AB - Samples of the microbial community from Lake Michie, a mesotrophic reservoir in central North Carolina, were adapted to various levels (100 to 1,000 mug/liter) of natural humic acids in chemostats. The humic acids were extracted from water samples from Black Lake, a highly colored lake in the coastal plain of North Carolina. After adaptation, the microbial community was tested for its ability to degrade the monosubstituted phenols m-cresol, m-aminophenol, and p-chlorophenol. Adaptation to increasing levels of humic acids significantly reduced the ability of the microbial communities to degrade all three phenols. The decline in biodegradation was accompanied by a decrease in the number of specific compound degraders in the adapted communities. Short-term exposure of the community to increasing levels of humic acids had no significant effect on the ability of the community to degrade m-cresol. Thus the suppressive effect of humic acids on monosubstituted phenol metabolism was the result of long-term exposure to the humic materials. Increasing the levels of inorganic nutrients fed to the chemostats during the humic acid adaptation had little effect on the suppressive influence of the humic acids, indicating that nutrient limitation was probably not responsible for the metabolic suppression. The results of the study suggest that long-term exposure to humic acids can reduce the ability of microbial communities to respond to monosubstituted phenols. PMID- 16346727 TI - Microcalorimetric Approach to Determine Relationships between Energy Supply and Metabolism in River Epilithon. AB - In a study to determine the relative importance of various apparent molecular weight fractions to the metabolism (heat output) of attached microorganisms in rivers, a marked lack of response to gross changes in the external organic matter supply was noted over the course of several hours. Such a response by a mixed autotrophic-heterotrophic community could be attributed to the autotrophs being the sole organic energy source for the heterotrophs. However, substantial metabolic activities were noted in attached microbial communities grown in darkness (algae free), thus indicating that riverborne organic matter was a major energy source. This paradox was attributed to a proposed lag in the availability of dissolved and colloidal organic matter to the heterotrophic microorganisms. PMID- 16346728 TI - Effect of drying medium on residual moisture content and viability of freeze dried lactic Acid bacteria. AB - The effect of various substances on the relationship between residual moisture content and the viability of freeze-dried lactic acid bacteria has been studied. Compounds such as polymers, which display considerable ability in displacing water, showed no protective action during freeze-drying. Adonitol, on the other hand, produced the smallest change in water content at various times during drying and allowed the highest rate of survival. PMID- 16346729 TI - Population Dynamics of Soil Pseudomonads in the Rhizosphere of Potato (Solanum tuberosum L.). AB - Rhizosphere population dynamics of seven Pseudomonas fluorescens and Pseudomonas putida strains isolated from rhizospheres of various agricultural plants were studied on potato (Solanum tuberosum L.) in field soil under controlled environmental conditions. Rhizosphere populations of two strains (B10 and B4) were quantitatively related to initial seed piece inoculum levels when plants were grown at -0.3 bar matric potential. At a given inoculum level, rhizosphere populations of strain B4 were consistently greater than those of strain B10. In vivo growth curves on 4-cm root tip-proximal segments indicated that both strains grew at similar rates in the potato rhizosphere, but large populations of strain B10 were not maintained at 24 degrees C after 7 h, whereas those of strain B4 were maintained for at least 40 h. Although both strains grew more rapidly in the rhizosphere at 24 degrees C than at 12 degrees C, their rhizosphere populations after seed piece inoculation were generally greater at 12 or 18 degrees C, indicating that in vivo growth did not solely determine rhizosphere populations in these studies. In vitro osmotolerance of seven Pseudomonas strains (including strains B4 and B10) was correlated with their abilities to establish stable populations in the rhizosphere of potato. Stability of rhizosphere populations of the Pseudomonas strains studied here was maximized at low (i.e., 12 degrees C) soil temperatures. These results indicate that Pseudomonas strains differ in their capacity to maintain stable rhizosphere populations in association with potato. This capacity, distinct from the ability to grow in the rhizosphere, may limit the establishment of rhizosphere populations under some environmental conditions. PMID- 16346730 TI - Straw and Xylan Utilization by Pure Cultures of Nitrogen-Fixing Azospirillum spp. AB - Azospirillum spp. were shown to utilize both straw and xylan, a major component of straw, for growth with an adequate combined N supply and also under N-limiting conditions. For most strains examined, a semisolid agar medium was satisfactory, but several strains appeared to be capable of slow metabolism of the agar. Subsequently, experiments were done with acid-washed sand supplemented with various carbon sources. In these experiments, authenticated laboratory strains, and all 16 recent field isolates from straw-amended soils, of both A. brasilense and A. lipoferum possessed the ability to utilize straw and xylan as energy sources for nitrogen fixation. Neither carboxymethyl cellulose nor cellulose was utilized. The strains and isolates differed in their abilities to utilize xylan and straw and in the efficiency of nitrogenase activity (CO(2)/C(2)H(2) ratio). Reasonable levels of activity could be maintained for at least 14 days in the sand cultures. Nitrogenase activity (acetylene reduction) was confirmed by N(2) incorporation. The level of nitrogenase activity observed was dependent on the time of the addition of acetylene to the culture vessels. PMID- 16346731 TI - Degradation of the herbicide mecoprop [2-(2-methyl-4-chlorophenoxy)propionic Acid] by a synergistic microbial community. AB - A microbial community isolated from wheat root systems was capable of growth on mecoprop as the sole carbon and energy source. When exposed to fresh herbicide additions, the community was able to shorten the lag phase from 30 days to less than 24 h. The community comprised two Pseudomonas species, an Alcaligenes species, a Flavobacterium species, and Acinetobacter calcoaceticus. None of the pure cultures was capable of growing on mecoprop. Certain combinations of two or more community constituents were required before growth commenced. The mecoprop degrading community could also degrade 2,4-dichlorophenoxyacetic acid and 2 methyl-4-chlorophenoxyacetic acid but not 2,4,5-trichlorophenoxyacetic acid. PMID- 16346732 TI - Kinetic studies of bacterial sulfate reduction in freshwater sediments by high pressure liquid chromatography and microdistillation. AB - Indirect photometric chromatography and microdistillation enabled a simultaneous measurement of sulfate depletion and sulfide production in the top 3 cm of freshwater sediments to be made. The simultaneous measurement of sulfate depletion and sulfide production rates provided added insight into microbial sulfur metabolism. The lower sulfate reduction rates, as derived from the production of acid-volatile S only, were explained by a conversion of this pool to an undistillable fraction under acidic conditions during incubation. A mathematical model was applied to calculate sulfate reduction from sulfate gradients at the sediment-water interface. To avoid disturbance of these gradients, the sample volume was reduced to 0.2 g (wet weight) of sediment. Sulfate diffusion coefficients in the model were determined (D(s) = 0.3 x 10 cm s at 6 degrees C). The results of the model were compared with those of radioactive sulfate turnover experiments by assessing the actual turnover rate constants (2 to 5 day) and pool sizes of sulfate at different sediment depths. PMID- 16346733 TI - Fruiting Body Formation from Regenerated Mycelium of Pleurotus ostreatus Protoplasts. AB - Fruiting bodies were induced from mycelium regenerated from Pleurotus ostreatus protoplasts. Mycelia originated from protoplasts conformed to parental strain mycelia in morphology. Six strains selected at random from the dikaryotic regenerants were able to form normal fruiting bodies, yielding 18% more than the parent. PMID- 16346734 TI - Activated sludge biodegradation of 12 commercial phthalate esters. AB - The activated sludge biodegradability of 12 commercial phthalate esters was evaluated in two test systems: (i) a semicontinuous activated sludge test and (ii) an acclimated 19-day die-away procedure. Both procedures demonstrated that phthalate esters are rapidly biodegraded under activated sludge conditions when loss of the parent phthalate ester (primary degradation) is measured. PMID- 16346735 TI - Improved fixation of cellulose-acetate reverse-osmosis membrane for scanning electron microscopy. AB - Fixation of cellulose-acetate membranes with either glutaraldehyde-osmium tetroxide or glutaraldehyde-ruthenium tetroxide resulted in extensive electron beam damage. Beam damage was eliminated and the bacterial surface structure was preserved, however, when cellulose-acetate membranes were fixed with glutaraldehyderuthenium tetroxide and treated successively with thiocarbohydrazide and osmium tetroxide. PMID- 16346736 TI - Possible Involvement of Phage-Like Structures in Antagonism of Cowpea Rhizobia by Rhizobium trifolii. AB - A reduction in the viability of cowpea rhizobia was observed when Rhizobium trifolii IARI and cowpea Rhizobium strain 3824 were inoculated together in soil. The reduction in number of cowpea rhizobia in soil was found to be associated with the reduction in number of nodules per plant and retardation in plant growth. An antimicrobial substance was isolated from R. trifolii which, on electron microscopic investigation, demonstrated the presence of several phage like structures. PMID- 16346737 TI - Quantitative addition of dissolved oxygen to in situ benthic chamber systems by use of catalase and hydrogen peroxide. AB - A methodology for reoxygenation of in situ benthic chamber systems by enzymatic catalysis of hydrogen peroxide with catalase was developed. For a 10-liter benthic chamber, the injection of 1 ml of catalase suspension (26,000 U ml) followed by 10 ml of 0.5 M hydrogen peroxide solution resulted in complete reoxygenation within 2.5 min at 25 degrees C. PMID- 16346738 TI - Inoculant maturity influences survival of rhizobia on seed. AB - Survival of Rhizobium trifolii on seeds of arrowleaf clover (Trifolium versiculosum Savi) and subclover (Trifolium subterraneum L.) was affected by the maturity of peat-, vermiculite-, and charcoal-based inoculants. Ten times more rhizobia survived on seed 4 days after inoculation when inoculants were stored (cured) before being utilized as compared with uncured inoculants. Increasing the curing time of inoculants beyond 4 weeks had little effect on increasing survival of seed-applied rhizobia. PMID- 16346739 TI - Construction of tn3-containing plasmids from plant-pathogenic pseudomonads and an examination of their biological properties. AB - Indigenous plasmids isolated from Pseudomonas tabaci ATCC 11528(pJP1), Pseudomonas angulata 45(pJP30), and P. tabaci BR2(pBPW1) (M. Obukowicz and P. D. Shaw, J. Bacteriol. 155:438-442, 1983) were labeled with Tn3, and the strains were subsequently cured of their respective plasmids. Plasmid-containing and cured isolates caused plant symptoms that were nearly indistinguishable, and the same amount of tabtoxin was produced by P. tabaci strains ATCC 11528 and BR2. PMID- 16346740 TI - Characteristics and Adaptability of Some New Isolates of Clostridium thermocellum. AB - Six strains of Clostridium thermocellum isolated from various environments were characterized as to growth rate, production of reducing sugars, ethanol, and acetic acid from cellulose, base composition of DNA, and the abilities to adapt to ethanol and to grow at 45 degrees C. Five of the six new isolates produced 7 to 15% more ethanol and two produced about 45% more reducing sugars than a standard reference strain. One strain (MC-6) adapted more readily to growth in 2% ethanol than the others. PMID- 16346741 TI - Decomposition of [C]Lignocelluloses of Spartina alterniflora and a Comparison with Field Experiments. AB - Decomposition of lignocelluloses from Spartina alterniflora in salt-marsh sediments was measured by using C-labeled compounds. Rates of decomposition were fastest in the first 4 days of incubation and declined later. Lignins labeled in side chains were mineralized slightly faster than uniformly labeled lignins; 12% of the [side chain-C]lignin-labeled lignocellulose was mineralized after 816 h of incubation, whereas only 8% of the [U-C]lignin-labeled lignocelluloses were degraded during this period. The carbohydrate moiety within the lignocellulose complex was degraded about four times faster than the lignin moiety; after 816 h of incubation, 29 to 37% of the carbohydrate moiety had been mineralized. Changes in concentration of lignin and cellulose in litter of S. alterniflora were followed over 2 years of decay. Cellulose disappeared from litter more rapidly than lignin; 50% of the initial content of cellulose was lost after 130 days, whereas lignin required 330 to 380 days for 50% loss. The slow loss of lignin compared with other litter components resulted in a progressive enrichment of litter in lignin content. The rates of mineralization of [C]lignocelluloses in marsh sediments were similar to the rates of lignocellulose decomposition in litter on the marsh. PMID- 16346742 TI - Primary and bacterial production in two dimictic indiana lakes. AB - The relationship between primary and bacterial production in two dimictic Indiana lakes with different primary productivities was examined during the summer stratification period in 1982. Primary production rates were calculated from rates of HCO(3) incorporation by natural samples, and bacterial production was calculated from rates of [H-methyl]thymidine incorporation by natural samples. Both vertical and seasonal distributions of bacterial production in the more productive lake (Little Crooked Lake) were strongly influenced by primary production. A lag of about 2 weeks between a burst in primary production and the subsequent response in bacterial production was observed. The vertical distribution of bacterial production in the water column of the less productive lake (Crooked Lake) was determined by the vertical distribution of primary production, but no clear relationship between seasonal maxima of primary and bacterial production in this lake was observed. High rates of bacterial production in Crooked Lake during May indicate the importance of allochthonous carbon washed in by spring rains. Bacterial production accounted for 30.6 and 31.8% of total (primary plus bacterial) production in Crooked Lake and Little Crooked Lake, respectively, from April through October. High rates of bacterial production during late September and October were observed in both lakes. Calculation of the fraction of bacterial production supported by phytoplankton excretion implies an important role for other mechanisms of supplying carbon, such as phytoplankton autolysis. Several factors affecting the calculation of bacterial production from the thymidine incorporation rates in these lakes were examined. PMID- 16346743 TI - Seasonal bacterial production in a dimictic lake as measured by increases in cell numbers and thymidine incorporation. AB - Rates of primary and bacterial production in Little Crooked Lake were calculated from the rates of incorporation of HCO(3) and [methyl-H]thymidine, respectively. Growth rates of bacteria in diluted natural samples were determined for epilimnetic and metalimnetic bacterial populations during the summers of 1982 and 1983. Exponential growth was observed in these diluted samples, with increases in cell numbers of 30 to 250%. No lag was observed in bacterial growth in 14 of 16 experiments. Correlation of bacterial growth rates to corresponding rates of thymidine incorporation by natural samples produced a conversion factor of 2.2 x 10 cells produced per mole of thymidine incorporated. The mass of the average bacterial cell in the lake was 1.40 x 10 +/- 0.05 x 10 g of C cell. Doubling times of natural bacteria calculated from thymidine incorporation rates and in situ cell numbers ranged from 0.35 to 12.00 days (median, 1.50 days). Bacterial production amounted to 66.7 g of C m from April through September, accounting for 29.4% of total (primary plus bacterial) production during this period. The vertical and seasonal distribution of bacterial production in Little Crooked Lake was strongly influenced by the distribution of primary production. From April through September 1983, the depth of maximum bacterial production rates in the water column was related to the depth of high rates of primary production. On a seasonal basis, primary production increased steadily from May through September, and bacterial production increased from May through August and then decreased in September. PMID- 16346744 TI - Thymidine incorporation by free-living and particle-bound bacteria in a eutrophic dimictic lake. AB - The percentage of [methyl-H]thymidine incorporated into samples from a dimictic eutrophic lake and retained on polycarbonate membranes of 3.0-, 1.0-, and 0.2-mum pore size was studied in a lake with filamentous cyanobacteria as the dominant phytoplankton type throughout the period of thermal stratification. Water samples were also examined by epifluorescence microscopy for evidence of algal senescence and bacterial colonization of intact and damaged cyanobacterial filaments. A small percentage (2 to 20%) of bacterial activity was retained by filters with pore sizes >/= 1 mum in epilimnetic samples. Epilimnetic samples also had a small percentage of cyanobacterial filaments, either intact or damaged, which were visibly colonized by bacteria in summer and fall samples. A significant proportion (20 to 35%) of bacterial activity was retained by filters with pore sizes >/= 1 mum in samples collected from the metalimnion and hypolimnion during late summer and fall. The proportion of damaged cyanobacterial filaments was higher in these samples than in those from the epilimnion or from those obtained early in the summer. Furthermore, the filaments in these samples were more heavily colonized by bacteria. Overall, particle-bound production accounted for only 2 to 19% of total bacterial production from April to August in all water layers. It appears that the supply of colonizable particles (damaged cyanobacterial filaments) is an important factor affecting the level of particle bound bacterial activity in this lake. PMID- 16346745 TI - Kinetics of denitrifying growth by fast-growing cowpea rhizobia. AB - Two fast-growing strains of cowpea rhizobia (A26 and A28) were found to grow anaerobically at the expense of NO(3), NO(2), and N(2)O as terminal electron acceptors. The two major differences between aerobic and denitrifying growth were lower yield coefficients (Y) and higher saturation constants (K(s)) with nitrogenous oxides as electron acceptors. When grown aerobically, A26 and A28 adhered to Monod kinetics, respectively, as follows: K(s), 3.4 and 3.8 muM; Y, 16.0 and 14.0 g . cells eq; mu(max), 0.41 and 0.33 h. Yield coefficients for denitrifying growth ranged from 40 to 70% of those for aerobic growth. Only A26 adhered to Monod kinetics with respect to growth on all three nitrogenous oxides. The apparent K(s) values were 41, 270, and 460 muM for nitrous oxide, nitrate, and nitrite, respectively; the K(s) for A28 grown on nitrate was 250 muM. The results are kinetically and thermodynamically consistent in explaining why O(2) is the preferred electron acceptor. Although no definitive conclusions could be drawn regarding preferential utilization of nitrogenous oxides, nitrite was inhibitory to both strains and effected slower growth. However, growth rates were identical (mu(max), 0.41 h) when A26 was grown with either O(2) or NO(3) as an electron acceptor and were only slightly reduced when A28 was grown with NO(3) (0.25 h) as opposed to O(2) (0.33 h). PMID- 16346746 TI - Modulation of acetone-butanol-ethanol fermentation by carbon monoxide and organic acids. AB - Metabolic modulation of acetone-butanol-ethanol fermentation by Clostridium acetobutylicum with carbon monoxide (CO) and organic acids is described. CO, which is a known inhibitor of hydrogenase, was found to be effective in the concentration range of dissolved CO corresponding to a CO partial pressure of 0.1 to 0.2 atm. Metabolic modulation by CO was particularly effective when organic acids such as acetic and butyric acids were added to the fermentation as electron sinks. The uptake of organic acids was enhanced, and increases in butyric acid uptake by 50 to 200% over control were observed. Hydrogen production could be reduced by 50% and the ratio of solvents could be controlled by CO modulation and organic acid addition. Acetone production could be eliminated if desired. Butanol yield could be increased by 10 to 15%. Total solvent yield could be increased 1 to 3% and the electron efficiency to acetone-butanol-ethanol solvents could be increased from 73 to 78% for controls to 80 to 85% for CO- and organic acid modulated fermentations. Based on these results, the dynamic nature of electron flow in this fermentation has been elucidated and mechanisms for metabolic control have been hypothesized. PMID- 16346747 TI - Bacterial Alginate Produced by a Mutant of Azotobacter vinelandii. AB - The optimum conditions in shaken flasks for production of bacterial alginate by mutant C-14 of Azotobacter vinelandii NCIB 9068 and a comparison of the properties of bacterial and algal alginates were investigated. The largest amount of bacterial alginate was obtained in about 110 h by a culture grown on optimum medium at 34 degrees C and 170-rpm shaking speed. The viscosity of the culture broth was 18,400 cps and the alginate concentration reached 6.22 g/liter. The viscosity of the purified bacterial alginate was as high as 11,200 cps at a low concentration (0.6%). A greater than fivefold concentration of algal alginate was required to reach the same viscosity at a low shear rate. A solution of bacterial alginate was more pseudoplastic than that of algal alginate was. No significant differences were observed in other properties of bacterial and algal alginates such as gel formation with calcium ion, thermostability, and effect of temperature, pH, and sodium chloride on viscosity. PMID- 16346748 TI - Occurrence of zearalenols (diastereomeric mixture) in corn stalk rot and their production by associated fusarium species. AB - Zearalenol was extracted from Fusarium-infected stems of corn from southern Italy. The toxin, which appeared as a single compound in various thin-layer chromatography systems, was resolved by high-pressure liquid chromatography into two components. A gas chromatography-mass spectrometry examination of a purified fraction confirmed the natural occurrence of zearalenol as a diastereomeric mixture and led to the identification of alpha (56 ng/g) and beta (27 ng/g) isomers. Among nine Fusarium species found associated with stalk rot in corn, only Fusarium culmorum (F. roseum ;Culmorum') and F. equiseti (F. roseum ;Gibbosum') produced zearalenol and always produced it in a diastereomeric mixture of alpha and beta isomers. PMID- 16346749 TI - Differential Adhesion and Infection of Nematodes by the Endoparasitic Fungus Meria coniospora (Deuteromycetes). AB - The conidia of the endoparasitic fungus Meria coniospora (Deuteromycetes) had different patterns of adhesion to the cuticles of the several nematode species tested; adhesion in some species was only to the head and tail regions, on others over the entire cuticle, whereas on others there was a complete lack of adhesion. After adhesion, the fungus usually infected the nematode. However, adhesion to third-stage larvae of five animal parasitic nematodes, all of which carry the cast cuticle from the previous molt, did not result in infection. M. coniospora infected animal parasitic nematodes when this protective sheath was removed. Seven preparations of sialic acid (N-acetylneuraminic acid) gave three types of response in adhesion-infection of nematodes: (i) a significant reduction in conidial adhesions; (ii) no interference with adhesion, but a 10-day delay in infection; and (iii) a delay in infection by 2 to 3 days. The current results support previous findings indicating involvement of sialic acids localized on nematode cuticles in recognition of prey by M. coniospora. PMID- 16346750 TI - Influence of Manganese on Growth of a Sheathless Strain of Leptothrix discophora. AB - Mn exerted various effects on the growth of Leptothrix discophora strain SS-1 in batch cultures depending on the concentration added to the medium. Concentrations of 0.55 to 5.5 muM Mn, comparable to those in the environment from which strain SS-1 was isolated, decreased cell yield and prolonged stationary-phase survival, but did not affect growth rate. Elevated concentrations of 55 to 910 muM Mn also decreased cell yield and prolonged survival, but growth rate was decreased as well. The addition of 1,820 muM Mn caused a decline in cell numbers followed by an exponential rise after 80 h of incubation, indicating the development of a population of cells resistant to Mn toxicity. When 360 muM Mn or less was added to growth flasks, Mn was oxidized to manganese oxide (MnO(x), where x is approximately 2), which appeared as brown particles in the medium. Quantification of Mn oxidation during growth of cultures to which 55 muM Mn was added showed that nearly all of the Mn was oxidized by the beginning of the stationary phase of growth (15 to 25 h). This result suggested that the decrease in cell yield observed at low and moderate concentrations of Mn was related to the formation of MnO(x), which may have bound cationic nutrients essential to the growth of SS-1. The addition of excess Fe to cultures containing 55 muM Mn increased cell yield to levels near those found in cultures with no added Mn, indicating that iron deprivation by MnO(x) was at least partly responsible for the decreased cell yield. PMID- 16346751 TI - Insect feeding deterrents in endophyte-infected tall fescue. AB - The presence of an endophytic fungus, Acremonium coenophialum, in tall fescue (Festuca arundinacea) deterred aphid feeding by Rhopalosiphum padi and Schizaphis graminum. Both species of aphid were unable to survive when confined to endophyte infected tall fescue plants. Feeding deterrents and toxic factors to R. padi and Oncopeltus fasciatus, large milkweed bug, were primarily associated with a methanol extract obtained when endophyte-infected tall fescue seed was serially extracted with hexane, ethyl acetate, and methanol. The concentrations of pyrrolizidine alkaloids were determined to be 30 to 100 times greater in the methanol extract than in the hexane and ethyl acetate extracts. PMID- 16346752 TI - Anaerobic Degradation of Carrageenan from the Red Macroalga Eucheuma cottonii. AB - Anaerobic degradation of the sulfated polysaccharide carrageenan was investigated by batch digestion of the red macroalga Eucheuma cottonii. During a 10-week incubation, ca. 60% of the starting E. cottonii biomass was fermented to CO(2), methane, and volatile fatty acids (predominantly acetate). Carrageenan degradation paralleled the loss of total biomass, suggesting no preferential degradation or preservation. After 10 weeks of incubation, the carrageenan content of the remaining biomass was 51%, as opposed to 61% of the original E. cottonii biomass. Carrageenan recovered after 10 weeks of digestion had a lower average molecular weight (319,000 versus 510,000) and formed solutions with considerably lower viscosities than did intact carrageenan. The percent C and percent N content of the particulate material in the digestors increased over time, probably as a result of microbial growth. In contrast, the percent S content decreased continuously; the loss of sulfur was most likely a result of the hydrolysis of carrageenan. Results from this study indicate that it is not economically viable to process E. cottonii simultaneously for hydrocolloids and methane. PMID- 16346753 TI - Methanosarcina mazei LYC, a New Methanogenic Isolate Which Produces a Disaggregating Enzyme. AB - A methanogenic coccoid organism, Methanosarcina mazei LYC, was isolated from alkaline sediment obtained from an oil exploration drilling site. The isolate resembled M. mazei S-6 by exhibiting different morphophases during its normal growth cycle. It differed from M. mazei S-6 by undergoint a spontaneous shift from large, irregular aggregates of cells to small, individual, irregular, coccoid units. In batch cultures at pH 7.0, M. mazei LYC grew as aggregates during the early growth stage. As the batch culture began exponential growth, the cell aggregates spontaneously dispersed: the culture liquid became turbid, and myriads of tiny (diameter, 1 to 3 mum) coccoid units were observed under phase contrast microscopy. Disaggregation apparently was accomplished by the production of an enzyme which hydrolyzed the heteropolysaccharide component of the cell wall; the enzyme was active on other Methanosarcina strains as well. Although the enzyme was active when tested at pH 6.0, it apparently was not produced at that pH: when strain LYC was grown at pH 6.0, only cell aggregates were present throughout batch growth. Individual coccoid cells of M. mazei LYC were sensitive to sodium dodecyl sulfate, but the large aggregates of cells were not. Strain LYC rapidly used H(2)-CO(2), in addition to methanol, and mono-, di-, and trimethylamine as methanogenic substrates; acetate was used very slowly. Its optimum growth temperature was 40 degrees C, and its optimum pH was 7.2. PMID- 16346754 TI - Nutrition and Growth Characteristics of Trichomitopsis termopsidis, a Cellulolytic Protozoan from Termites. AB - Putatively axenic cultures of Trichomitopsis termopsidis 6057, isolated by M. A. Yamin (J. Protozool. 25:535-538, 1978) from the hindgut of Zootermopsis termites, apparently contained methanogenic bacteria, inasmuch as small amounts of CH(4) were produced during growth. However, T. termopsidis could be "cured" of methanogenic activity by incubation in the presence of bromoethanesulfonate. Both the cured derivative (6057C) and the parent strain (6057) required NaHCO(3) and fetal bovine serum for good growth; the presence of yeast extract in media was stimulatory. Growth of both strains was markedly improved by substituting heat killed cells of Bacteroides sp. strain JW20 (a termite gut isolate) for heat killed rumen bacteria in media as a source of bacterial cell material. Heat killed Bacteroides sp. strain JW20 was the best of a number of bacteria tested, and under these conditions H(2) was a major protozoan fermentation product. Growth of T. termopsidis strains was further improved by co-cultivation in the presence of Methanospirillum hungatii. M. hungatii was the best of a number of H(2)-consuming bacteria tested, and under these conditions CH(4), but not H(2), was produced, indicating interspecies transfer of H(2) between the protozoa and M. hungatii. Both strains of T. termopsidis used powdered, particulate forms of cellulose (e.g., pure cellulose, corncob, cereal leaves) as fermentable energy sources, although powdered wood, chitin, or xylan supported little or no growth. Cells of the cellulose-forming coccus Sarcina ventriculi also served as a fermentable energy source, but these were used poorly as a source of bacterial cell material. The only substantial difference between T. termopsidis 6057 and 6057C was that the latter grew poorly or not at all with rumen bacteria as a source of bacterial cell material. The improved growth of T. termopsidis in vitro should facilitate further studies on the cell biology and biochemistry of these symbiotic, anaerobic protozoa. PMID- 16346755 TI - Cellulase and Other Polymer-Hydrolyzing Activities of Trichomitopsis termopsidis, a Symbiotic Protozoan from Termites. AB - Crude extracts of the anaerobic, cellulolytic protozoan Trichomitopsis termopsidis possessed endo-beta-1,4-glucanase and cellobiase activities, as evidenced by hydrolytic action on carboxymethyl cellulose and cellobiose, respectively. Cell extracts also hydrolyzed microcrystalline cellulose. Hydrolysis of microcrystalline cellulose displayed optima at pH 5 and at 30 degrees C, and glucose was the sole product liberated. Cellulolytic activities of T. termopsidis appeared to be entirely cell associated. Hydrolytic activity was also detected against Douglas fir wood powder, xylan, starch, and protein, but not chitin. The importance of these enzymes in the nutrition of T. termopsidis is discussed in terms of the natural habitat of this protozoan (the hindgut of wood eating termites). PMID- 16346756 TI - Transfer of Sucrose-Fermenting Ability and Nisin Production Phenotype among Lactic Streptococci. AB - Transfer of sucrose fermentation ability, nisin production, and nisin resistance from Streptococcus lactis to S. lactis and Streptococcus lactis subsp. diacetylactis occurred between cells immobilized on nitrocellulose filters in the presence of DNase. Transconjugants were able to act as donors to transfer the Suc Nis phenotype in subsequent mating. No changes in sensitivity to lytic phage c2 were noted in S. lactis transconjugants. However, temperature-independent restriction of lytic phage 18-16 was noted in transconjugants of S. lactis subsp. diacetylactis 18-16. Adsorption studies with phage-resistant transconjugants showed that resistance was not due to lack of adsorption by the lytic phage. Physical evidence for the presence of introduced plasmid DNA was not found in lysates of transconjugants. PMID- 16346757 TI - Annual cycle of bacterial secondary production in five aquatic habitats of the okefenokee swamp ecosystem. AB - Rates of bacterial secondary production by free-living bacterioplankton in the Okefenokee Swamp are high and comparable to reported values for a wide variety of marine and freshwater ecosystems. Bacterial production in the water column of five aquatic habitats of the Okefenokee Swamp was substantial despite the acidic (pH 3.7), low-nutrient, peat-accumulating character of the environment. Incorporation of [H]thymidine into cold-trichloroacetic acid-insoluble material ranged from 0.03 to 2.93 nmol liter day) and corresponded to rates of bacterial secondary production of 3.4 to 342.2 mug of carbon liter day (mean, 87.8 mug of carbon liter day). Bacterial production was strongly seasonal and appeared to be coupled to annual changes in temperature and primary production. Bacterial doubling times ranged from 5 h to 15 days and were fastest during the warm months of the year, when the biomass of aquatic macrophytes was high, and slowest during the winter, when the plant biomass was reduced. The high rates of bacterial turnover in Okefenokee waters suggest that bacterial growth is an important mechanism in the transformation of dissolved organic carbon into the nutrient rich bacterial biomass which is utilized by microconsumers. PMID- 16346758 TI - Differences in Xylan Degradation by Various Noncellulolytic Thermophilic Anaerobes and Clostridium thermocellum. AB - Hemicellulose fractions with a predetermined distribution of xylose, xylooligomers, and xylan fractions were obtained through steam explosion of wood by the steam explosion-extraction process of BFA-Hamburg, Hamburg, Federal Republic of Germany. A differential utilization of various molecular-weight fractions by several thermophilic anaerobic bacteria was determined during their growth on the hemicellulose preparations. Clostridium thermocellum (60 degrees C) first utilized the high-molecular-weight fractions (polymerization degree of 15 to 40 xylose units). Xylose and xylooligomers of n = 2 to 5 accumulated while C. thermocellum was not growing, as evident from the fermentation products formed. Whereas the xylan was hydrolyzed and the small oligoxylans were utilized after more than 100 h of incubation, xylose was not significantly utilized. In contrast to this, C. thermohydrosulfuricum (70 degrees C) and Thermoanaerobium brockii (70 degrees C) utilized xylose first and then xylooligomers of n = 2 to 5, but xylooligomers of n greater than 6 were only slowly utilized. Thermoanaerobacter ethanolicus (70 degrees C), Thermobacteroides acetoethylicus (70 degrees C), and C. thermosaccharolyticum (60 degrees C) utilized xylose preferentially. Xylooligomers of n = 2 to 5 and n = 6 and greater were apparently concomitantly utilized without significant differences. In contrast to C. thermocellum, the non cellulolytic organisms grew during xylan hydrolysis, producing ethanol, lactate, acetate, CO(2), and H(2). PMID- 16346759 TI - Role of Penicillic Acid in the Phytotoxicity of Penicillium cyclopium and Penicillium canescens to the Germination of Corn Seeds. AB - Penicillium cyclopium and Penicillium canescens cultures inhibited the germination of corn. The phytotoxic compound was isolated by solvent extraction and thin-layer chromatography on silica gel. The phytotoxin was identified as penicillic acid by mass spectrometry, nuclear magnetic resonance, and infrared spectroscopy. Gas-liquid chromatography on a capillary glass column separated the two epimeric forms of penicillic acid. The maximum production of penicillic acid was obtained with P. cyclopium cultures grown at 25 degrees C. The phytotoxicity of penicillic acid was manifested by its ability to alter the germination of corn. The percent inhibition of germination was directly proportional to the logarithm of the penicillic acid concentration. Growth of the main root was reduced 50% by concentrations of 500 mug/ml. PMID- 16346760 TI - Bacterial fouling in a model core system. AB - We have used a sintered glass bead core to simulate the spaces and surfaces of reservoir rock in studies of the bacterial plugging phenomenon that affects waterflood oil recovery operations. The passage of pure or mixed natural populations of bacteria through this solid matrix was initially seen to promote the formation of adherent bacterial microcolonies on available surfaces. Bacteria within these microcolonies produced huge amounts of exopolysaccharides and coalesced to form a confluent plugging biofilm that eventually caused a >99% decrease in core permeability. Aerobic bacteria developed a plugging biofilm on the inlet face of the core, facultative anaerobes plugged throughout the core, and dead bacteria did not effectively plug the narrow (33-mum) spaces of this solid matrix because they neither adhered extensively to surfaces nor produced the extensive exopolysaccharides characteristic of living cells. The presence of particles in the water used in these experiments rapidly decreased the core permeability because they became trapped in the developing biofilm and accelerated the plugging of pore spaces. Once established, cells within the bacterial biofilm could be killed by treatment with a biocide (isothiazalone), but their essentially inert carbohydrate biofilm matrix persisted and continued to plug the pore spaces, whereas treatment with 5% sodium hypochlorite killed the bacteria, dissolved the exopolysaccharide biofilm matrix, and restored permeability to these plugged glass bead cores. PMID- 16346762 TI - Modified agar medium for detecting environmental salmonellae by the most-probable number method. AB - [This corrects the article on p. 1027 in vol. 48.]. PMID- 16346761 TI - Influence of calcium addition on growth of highly purified syntrophic cultures degrading long-chain Fatty acids. AB - Two highly purified syntrophic associations resulting in acetogenesis from stearate (SM) and oleate (OM) were obtained from the sludges of a sewage digestor. In both cases, Methanospirillum hungatei together with short, motile, gram-negative, nonfluorescent rods morphologically similar to Syntrophomonas wolfei were identified by microscopic examination. Besides growing on volatile fatty acids (butyrate through caproate), both cultures grew on oleate (C(18:1)) and numerous even-numbered, saturated long-chain fatty acids (LCFA [decanoate through stearate]). In addition, during growth on LCFA, supplementation of the culture media with calcium chloride was an absolute requirement. The sole difference between the associations was observed when SM and OM cultures were transferred from a stearate to an oleate medium. The SM culture needed 10 days before starting to degrade oleate, whereas the OM culture grew immediately, but the OM culture also grew immediately when transferred to stearate medium. Saturated LCFA degradation occurred in the presence of equinormal amounts of calcium (fatty acid/Ca ratio, 2). On the other hand, OM degradation only took place in the presence of an equimolar amount of calcium (fatty acid/Ca ratio, 1). These observations are discussed by considering the solubility constants of LCFA as calcium salts and the toxicity of the free acids against microorganisms. PMID- 16346763 TI - General method for the identification of plasmid species in fast-growing soil microorganisms. AB - Using a horizontal gel electrophoresis method, we demonstrated reproducibly the presence of indigenous plasmids in different Rhizobium, Agrobacterium, and Pseudomonas strains. The method yields a large amount of plasmid DNA and is sensitive in detecting megaplasmids with molecular weights higher than 5 x 10. In two Rhizobium meliloti strains, a megaplasmid other than the low-mobility plasmid already known was detected. PMID- 16346764 TI - Isolation of Noninhibitory Strains of Zymomonas mobilis. AB - Wild-type Zymomonas mobilis strains inhibit the growth of Escherichia coli. We report the first isolation of noninhibitory strains, called Zymomonas inhibition negative (Zin), after treatment with N-methyl-N'-nitro-N-nitrosoguanidine. A standardized soft-agar overlay procedure for detecting E. coli growth inhibition was also developed. PMID- 16346765 TI - Simplified Enzyme-Linked Immunosorbent Assay for Routine Identification of Rhizobium japonicum Antigens. AB - A simple, reliable, and flexible modification of the indirect enzyme-linked immunosorbent assay was developed for the identification of Rhizobium japonicum antigens from cultures and nodules. The procedure emphasizes efficient use of time and reagents, adaptability to variously equipped laboratories, and maintenance of sensitivity levels that are adequate for ecological studies. PMID- 16346766 TI - Distribution of Alginate Lyase Activity among Strains of Bacillus circulans. AB - Strains from four different DNA relatedness groups of Bacillus circulans showed apparent alginate lyase activity; the activity of three strains examined had mannuronidase specificity. A representative strain of group 4 also produced apparent inducible unsulfated chrondroitin lyase activity. PMID- 16346767 TI - Anaerobic Growth and Denitrification among Different Serogroups of Soybean Rhizobia. AB - We screened soybean rhizobia originating from three germplasm collections for the ability to grow anaerobically in the presence of NO(3) and for differences in final product formation from anaerobic NO(3) metabolism. Denitrification abilities of selected strains as free-living bacteria and as bacteroids were compared. Anaerobic growth in the presence of NO(3) was observed in 270 of 321 strains of soybean rhizobia. All strains belonging to the 135 serogroup did not grow anaerobically in the presence of NO(3). An investigation with several strains indicated that bacteria not growing anaerobically in the presence of NO(3) also did not utilize NO(3) as the sole N source aerobically. An exception was strain USDA 33, which grew on NO(3) but failed to denitrify. Dissimilation of NO(3) by the free-living cultures proceeded without the significant release of intermediate products. Nitrous oxide reductase was inhibited by C(2)H(2), but preceding steps of denitrification were not affected. Final products of denitrification were NO(2), N(2)O, or N(2); serogroups 31, 46, 76, and 94 predominantly liberated NO(2), whereas evolution of N(2) was prevalent in serogroups 110 and 122, and all three were formed as final products by strains belonging to serogroups 6 and 123. Anaerobic metabolism of NO(3) by bacteroid preparations of Bradyrhizobium japonicum proceeded without delay and was evident by NO(2) accumulation irrespective of which final product was formed by the strain as free-living bacteria. Anaerobic C(2)H(2) reduction in the presence of NO(3) was observed in bacteroid preparations capable of NO(3) respiration but was absent in bacteria that were determined to be deficient in dissimilatory nitrate reductase. PMID- 16346768 TI - Simplified procedure for recovery of lignin acidolysis products for determining the lignin-degrading abilities of microorganisms. AB - A simplified procedure for the identification and measurement of single-ring aromatic products of lignin acidolysis is described. The procedure employed a 6-h hydrolysis of spruce milled wood lignin in acidic dioxane at 87 degrees C, followed by a series of organic extractions to recover acidolysis products which were quantified by gas chromatography of trimethylsilyl derivatives. Complex gel permeation chromatography procedures utilized by other workers were avoided in the modified procedure, but equivalent results were obtained. The simplified procedure was utilized to hydrolyze sound and actinomycete-decayed spruce milled wood lignins and was shown to be useful as a technique for the rapid screening of microorganisms for their ability to alter lignin. PMID- 16346769 TI - Studies on the Inoculation and Competitiveness of a Rhizobium leguminosarum Strain in Soils Containing Indigenous Rhizobia. AB - The competitiveness of a Rhizobium leguminosarum strain was investigated at two separate locations in field inoculation studies on commercially grown peas. The soil at each location (sites I and II) contained an indigenous R. leguminosarum population of ca. 3 x 10 rhizobia per g of soil. At site I it was necessary to use an inoculum concentration as large as 4 x 10 CFU ml (2 x 10 bacteria seed) to establish the inoculum strain in the majority of nodules (73%). However, at site II the inoculum strain formed only 33% of nodules when applied at this (10 CFU ml) level. Establishment could not be further improved by increasing the inoculum concentration even as high as 10 CFU ml (9.6 x 10 bacteria seed). The inoculum strain could be detected at both sites 19 months after inoculation. Analysis by intrinsic antibiotic resistance patterns and plasmid DNA profiles indicated that a dominant strain(s) and plasmid pool existed among the indigenous population at site II. Competition experiments were carried out under laboratory conditions between a dominant indigenous isolate and the inoculum strain. Both strains were shown to be equally competitive. PMID- 16346770 TI - Effect of Colloidal Materials on Cellulase Production by Trichoderma reesei Rut C30. AB - The addition of positively charged colloidal materials to the growth medium markedly increased the concentration of cellulase enzymes produced by Trichoderma reesei Rut-C30. Filter paper activities of up to 4 and 13 IU/ml have been achieved by the addition of colloidal materials, using 3% lactose and 3% cellulose, respectively, as a substrate. The particles exert their effect by binding soluble sugars and slowing their uptake by the organism. PMID- 16346771 TI - Nitrogen Fixation Associated with the New Zealand Mangrove (Avicennia marina (Forsk.) Vierh. var. resinifera (Forst. f.) Bakh.). AB - Nitrogenase activity in mangrove forests at two locations in the North Island, New Zealand, was measured by acetylene reduction and N(2) uptake. Nitrogenase activity (C(2)H(2) reduction) in surface sediments 0 to 10 mm deep was highly correlated (r = 0.91, n = 17) with the dry weight of decomposing particulate organic matter in the sediment and was independent of light. The activity was not correlated with the dry weight of roots in the top 10 mm of sediment (r = -0.01, n = 13). Seasonal and sample variation in acetylene reduction rates ranged from 0.4 to 50.0 mumol of C(2)H(4) m h under air, and acetylene reduction was depressed in anaerobic atmospheres. Nitrogen fixation rates of decomposing leaves from the surface measured by N(2) uptake ranged from 5.1 to 7.8 nmol of N(2) g (dry weight) h, and the mean molar ratio of acetylene reduced to nitrogen fixed was 4.5:1. Anaerobic conditions depressed the nitrogenase activity in decomposing leaves, which was independent of light. Nitrogenase activity was also found to be associated with pneumatophores. This activity was light dependent and was probably attributable to one or more species of Calothrix present as an epiphyte. Rates of activity were generally between 100 and 500 nmol of C(2)H(4) pneumatophore h in summer, but values up to 1,500 nmol of C(2)H(4) pneumatophore h were obtained. PMID- 16346772 TI - Influence of azospirillum strains on the nodulation of clovers by Rhizobium strains. AB - Mixed cultures of several Azospirillum and Rhizobium trifolii strains caused either an inhibition or stimulation of nodule formation on plant hosts as compared with nodulation of plants inoculated with R. trifolii alone. Azospirillum strains affected the nodulation process at a precise cell ratio (R. trifolii/Azospirillum cells) and time of inoculation. All Azospirillum strains used showed a variation in their ability to inhibit or enhance nodulation by R. trifolii strains. When nonviable cell preparations of Azospirillum strains were used for mixing experiments, no effect on nodulation was observed. A decrease in the effectiveness of normally Nod FixR. trifolii strains was observed when an Azospirillum strain caused an increase in nodule number. PMID- 16346773 TI - Interaction of azospirillum and Rhizobium strains leading to inhibition of nodulation. AB - Rhizobium-Azospirillum interactions during establishment of Rhizobium-clover symbiosis were studied. When mixed cultures of Azospirillum and Rhizobium trifolii strains were simultaneously inoculated onto clover plants, no nodulation by R. trifolii was observed. R. trifolii ANU1030, which nodulated clover plants without attacking root hairs, i.e., does not cause root hair curling (Hac), did not show inhibition of nodulation when inoculated together with Azospirillum strains. Isolation of bacteria from surface-sterilized roots showed that azospirilla could be isolated both from within root segments and from nodules. Inhibition of nodulation could be mimicked by the addition of auxins to the plant growth medium. PMID- 16346774 TI - Partial Purification and Characterization of a Polysaccharide Depolymerase Associated with Phage-Infected Erwinia amylovora. AB - Erwinia amylovora infected with bacteriophage ERA103 produced an enzyme which degraded the extracellular polysaccharide of noninfected cells. The depolymerase enzyme was purified 15-fold by a procedure which included ammonium sulfate precipitation, ultracentrifugation, CM-Sephadex batchwise separation, Sephadex G 50 column chromatography, and Sephacryl S-200 column chromatography. The enzyme had a molecular weight of approximately 21,000 and a pH optimum of 6.0. Activity was enhanced by supplements of 2-mercaptoethanol or dithiothreitol. PMID- 16346775 TI - Use of the mushroom tissue block rapid pitting test to detect brown blotch pathogens. AB - Isolates of an Arthrobacter species known to produce a positive host pathogenicity test were evaluated by the mushroom tissue block rapid pitting test. All 11 isolates of the Arthrobacter species yielded a positive test response. The time required for a positive test was the same for Pseudomonas tolaasi and the Arthrobacter species. PMID- 16346776 TI - Optimization of conditions for production of channel catfish ovary cells and channel catfish virus DNA. AB - Medium supplements were examined for their effect on the growth of channel catfish ovary cells. It was found that the usual serum supplement of 10% fetal calf serum could be successfully replaced with a combination of 5% fetal calf serum and a mixture of insulin, transferrin, and selenous acid. It was also found that these cells could be grown in a more efficient manner on microcarrier beads. This type of culture produced 14 times the number of cells per milliliter of total medium used compared with the usual tissue culture flasks used for cell growth. The microcarrier system also provided for greater production efficiency of DNA from channel catfish virus, a virus that infects this cell line. PMID- 16346777 TI - Spatial Distribution, Structure, Biomass, and Physiology of Microbial Assemblages across the Southern Ocean Frontal Zones during the Late Austral Winter. AB - We examined the spatial distributions of picoplankton, nanoplankton, and microplankton biomass and physiological state relative to the hydrography of the Southern Ocean along 90 degrees W longitude and across the Drake Passage in the late austral winter. The eastern South Pacific Ocean showed some large-scale biogeographical differences and size class variability. Microbial ATP biomass was greatest in euphotic surface waters. The horizontal distributions of microbial biomass and physiological state (adenylate energy charge ratio) coincided with internal currents (fronts) of the Antarctic Circumpolar Current. In the Drake Passage, the biological scales in the euphotic and aphotic zones were complex, and ATP, total adenylate, and adenylate energy charge ratio isopleths were compressed due to the extension of the sea ice from Antarctica and constriction of the Circumpolar Current through the narrow passage. The physiological state of microbial assemblages and biomass were much higher in the Drake Passage than in the eastern South Pacific Ocean. The temperature of Antarctic waters, not dissolved organic carbon, was the major variable controlling picoplankton growth. Estimates of picoplankton production based on ATP increments with time suggest that production under reduced predation pressure was 1 to 10 mug of carbon per liter per day. Our results demonstrate the influence of large-scale hydrographic processes on the distribution and structure of microplankton, nanoplankton, and picoplankton across the Southern Ocean. PMID- 16346778 TI - Transformation of Halogen-, Alkyl-, and Alkoxy-Substituted Anilines by a Laccase of Trametes versicolor. AB - The laccase of the fungus Trametes versicolor was able to polymerize various halogen-, alkyl-, and alkoxy-substituted anilines, showing substrate specificity similar to that of horseradish peroxidase, whereas the laccase of Rhizoctonia praticola was active only with p-methoxyaniline. The substrate specificities of the enzymes were determined by using gas chromatography to measure the decrease in substrate concentration during incubation. With p-chloroaniline as the substrate, the peroxidase and the Trametes laccase showed maximum activity near pH 4.2. The transformation of this substrate gave rise to a number of oligomers, ranging from dimers to pentamers, as determined by mass spectrometry. The product profiles obtained by high-pressure liquid chromatography were similar for the two enzymes. A chemical reaction was observed between p-chloroaniline and an enzymatically formed dimer, resulting in the formation of a trimer. All three enzymes oxidized p-methoxyaniline to 2-amino-5-p-anisidinobenzoquinone di-p methoxyphenylimine, but only the T. versicolor laccase and the peroxidase caused the formation of a pentamer (2,5-di-p-anisidinobenzoquinone di-p methoxyphenylimine). Our results demonstrate that in addition to horseradish peroxidase, a T. versicolor laccase can also polymerize aniline derivatives. PMID- 16346779 TI - Adaptation to High-Intensity, Low-Wavelength Light among Surface Blooms of the Cyanobacterium Microcystis aeruginosa. AB - Natural populations of the nuisance bloom cyanobacterium Microcystis aeruginosa obtained from the eutrophic Neuse River, N.C., revealed optimal chlorophyll a normalized photosynthetic rates and resistance to photoinhibition at surface photosynthetically active radiation (PAR) intensities. At saturating PAR levels these populations exhibited higher photosynthetic rates in quartz than in Pyrex vessels. Eucaryotic algal populations obtained from the same river failed to counteract photoinhibition. At saturating PAR levels, such populations generally yielded lower photosynthetic rates in quartz containers than they did in Pyrex containers. Cultivation of natural Microcystis populations under laboratory conditions led to physiologically distinct populations which had photoinhibitory characteristics similar to those of other cultured cyanobacterial and eucaryotic algae. Our findings indicate that (i) photosynthetic production among natural surface populations is best characterized and quantified in quartz rather than Pyrex incubation vessels; (ii) extrapolation of natural photoinhibitory trends from laboratory populations is highly subjective to culture and PAR histories and may yield contradictory results; and (iii) buoyant surface-dwelling populations, rather than exhibiting senescence, are poised at optimizing PAR utilization, thereby maintaining numerical dominance in eutrophic waters when physico-chemical conditions favor bloom formation. PMID- 16346780 TI - Adaptation of Denitrifying Populations to Low Soil pH. AB - Natural denitrification rates and activities of denitrifying enzymes were measured in an agricultural soil which had a 20-year past history of low pH (pH ca. 4) due to fertilization with acid-generating ammonium salts. The soil adjacent to this site had been limed and had a pH of ca. 6.0. Natural denitrification rates of these areas were of similar magnitude: 158 ng of N g of soil day for the acid soil and 390 ng of N g of soil day at the neutral site. Estimates of in situ denitrifying enzyme activity were higher in the neutral soil, but substantial enzyme activity was also detected in the acid soil. Rates of nitrous oxide reduction were very low, even when NO(3) and NO(2) were undetectable, and were ca. 400 times lower than the rates of N(2)O production from NO(3). Denitrification rates measured in slurries of the acid and neutral soil showed distinctly different pH optima (pH 3.9 and pH 6.3) which were near the pH values of the two soils. This suggests that an acid-tolerant denitrifying population had been selected during the 20-year period of low pH. PMID- 16346781 TI - Biological and abiological sulfur reduction at high temperatures. AB - Reduction of elemental sulfur was studied in the presence and absence of thermophilic sulfur-reducing bacteria, at temperatures ranging from 65 to 110 degrees C, in anoxic artificial seawater media. Above 80 degrees C, significant amounts of sulfide were produced abiologically at linear rates, presumably by the disproportionation of sulfur. These rates increased with increasing temperature and pH and were enhanced by yeast extract. In the same medium, the sulfur respiration of two recent thermophilic isolates, a eubacterium and an archaebacterium, resulted in sulfide production at exponential rates. Although not essential for growth, sulfur increased the cell yield in both strains up to fourfold. It is suggested that sulfur respiration is favored at high temperatures and that this process is not limited to archaebacteria, but is shared by other extreme thermophiles. PMID- 16346782 TI - Inhibitory effect of pseudobactin on the uptake of iron by higher plants. AB - Purified pseudobactin inhibits the uptake of ferric iron by the roots of peas and maize plants sufficiently to reduce the synthesis of chlorophyll. This inhibition is interpreted as competitive binding, as described for synthetic chelating compounds. PMID- 16346783 TI - Test Medium for the Growth of Nitrosomonas europaea. AB - A mineral medium for studying the growth of Nitrosomonas europaea was developed and examined. The medium was defined in terms of chemical speciation by using chemical equilibrium computer models. The medium significantly increased the metabolic activity of the organisms compared with previously developed media, yielding a specific growth rate as high as 3.0 day (generation time, 5.5 h). The specific growth rate was enhanced by increasing the inoculum and was linearly correlated with the inoculum-to-total-culture volume ratio on a semilog scale. A reproducible growth rate for N. europaea was obtained with this medium under controlled experimental conditions. PMID- 16346784 TI - Impact of nitrogen and phosphorus on [C]lignocellulose decomposition by stream wood microflora. AB - Nutritional and physical factors affecting the decomposition of [C]lignocellulose prepared from Douglas fir (Pseudotsuga menziesii) were examined by incubating the labeled substrate with homogenized surface wood scrapings obtained from a Douglas fir log in a Pacific Northwest stream. Incubations were conducted in distilled water, in stream water collected from four different sources, or in a defined mineral salts solution with or without supplemental N (KNO(3)). Decomposition rates of [C]lignocellulose, as measured by CO(2) evolution, were greater in each of the four filter-sterilized sources of stream water than in distilled water alone. Decomposition experiments conducted in stream water media with the addition of defined mineral salts demonstrated that [C]cellulose decomposition was stimulated 50% by the addition of either KNO(3) or KH(2)PO(4)/K(2)HPO(4) and further enhanced (167%) by a combination of both. In contrast, [C]lignin decomposition was stimulated (65%) only by the addition of both N and P. Decomposition of [C]lignocellulose was greatest when supplemental KNO(3) was supplied in concentrations of at least 10.0 mg of N liter but not increased further by higher concentrations. The decomposition of [C]lignocellulose increased as the incubation temperature was raised and NO(3)-N supplementation further increased these rates between three-and sevenfold over the range of temperatures examined (5 to 22 degrees C). Accumulation of NH(4) (2 to 4 mg of N liter) was always observed in culture filtrates of incubations which had been supplemented with KNO(3), the quantity being independent of NO(3) concentrations >/= 10 mg of N liter. The role of supplemental NO(3) in the decomposition of [C]lignocellulose is discussed in relation to wood decomposition and the low concentrations of N found in stream ecosystems of the Pacific Northwest. PMID- 16346785 TI - Nitrogen dynamics in stream wood samples incubated with [C]lignocellulose and potassium [N]nitrate. AB - Surface wood samples obtained from a Douglas fir log (Pseudotsuga menziesii) in a Pacific Northwest stream were incubated in vitro with [C]lignocellulose in a defined mineral salts medium supplemented with 10 mg of N liter of N-labeled NO(3) (50 atom% N). Evolution of CO(2), distribution and isotopic dilution of N, filtrate N concentrations, and the rates of denitrification, N(2) fixation, and respiration were measured at 6, 12, and 18 days of incubation. The organic N content of the lignocellulose-wood sample mixture had increased from 132 mug of N to a maximum of 231 mug of N per treatment after 6 days of incubation. Rates of [C]lignocellulose decomposition were greatest during the first 6 days and then began to decline over the remaining 12 days. Total CO(2) evolution was also highest at day 6 and declined steadily over the remaining duration of the incubation. Filtrate NH(4)-N increased from background levels to a final value of 57 mug of N per treatment. Filtrate NO(3) N completely disappeared by day 6, and organic N showed a slight decline between days 12 and 18. The majority of the N that could be recovered appeared in the particulate organic fraction by day 6 (41 mug of N), and the filtrate NH(4) N fraction contained 11 mug of N by day 18. The N enrichment values of the filtrate NH(4) and the inorganic N associated with the particulate fraction had increased to approximately 20 atom% N by 18 days of incubation, whereas the particulate organic fraction reached its highest enrichment by day 6. Measurements of N(2) fixation and denitrification indicated an insignificant gain or loss of N from the experimental system by these processes. The data show that woody debris in stream ecosystems might function as a rapid and efficient sink for exogenous N, resulting in stimulation of wood decomposition and subsequent activation of other N cycling processes. PMID- 16346786 TI - Influence of Environmental Factors on Interstrain Competition in Rhizobium japonicum. AB - The effect of several biotic and abiotic factors on the pattern of competition between two strains of Rhizobium japonicum was examined. In two Minnesota soils, Waseca and Waukegan, strain USDA 123 occupied 69% (Waseca) and 24% (Waukegan) of the root nodules on Glycine max L. Merrill cv. Chippewa. USDA 110 occupied 2% of the root nodules in the Waseca soil and 12% of the nodules in the Waukegan soil. Under a variety of other growth conditions-vermiculite, vermiculite amended with Waseca soil, and two Hawaiian soils devoid of naturalized Rhizobium japonicum strains-USDA 110 was more competitive than USDA 123. The addition of nitrate to or the presence of antibiotic-producing actinomycetes in the rhizosphere of soybeans did not affect the pattern of competition between the two strains. However, preexposure of young seedings to USDA 110 or USDA 123 before transplantation into soil altered the pattern of competition between the two strains significantly. In the Waseca soil, preexposure of cv. Chippewa to USDA 110 for 72 h increased the percentage of nodules occupied by USDA 110 from 2 to 55%. Similarly, in the Hawaiian soil Waimea, nodule occupancy by USDA 123 increased from 7 to 33% after a 72-h preexposure. PMID- 16346787 TI - N Kinetic Analysis of N(2)O Production by Nitrosomonas europaea: an Examination of Nitrifier Denitrification. AB - A series of N isotope tracer experiments showed that Nitrosomonas europaea produces nitrous oxide only under oxygen-limiting conditions and that the labeled N from nitrite, but not nitrate, is incorporated into nitrous oxide, indicating the presence of the "denitrifying enzyme" nitrite reductase. A kinetic analysis of the m/z 44, 45, and 46 nitrous oxide produced by washed cell suspensions of N. europaea when incubated with 4 mM ammonium (99% N) and 0.4 mM nitrite (99% N) was performed. No labeled nitrite was reduced to ammonium. All labeled material added was accounted for as either nitrite or nitrous oxide. The hypothesis that nitrous oxide is produced directly from nitrification was rejected since (i) it does not allow for the large amounts of double-labeled (m/z 46) nitrous oxide observed; (ii) the observed patterns of m/z 44, 45, and 46 nitrous oxide were completely consistent with a kinetic analysis based on denitrification as the sole mechanism of nitrous oxide production but not with a kinetic analysis based on both mechanisms; (iii) the asymptotic ratio of m/z 45 to m/z 46 nitrous oxide was consistent with denitrification kinetics but inconsistent with nitrification kinetics, which predicted no limit to m/z 45 production. It is concluded that N. europaea is a denitrifier which, under conditions of oxygen stress, uses nitrite as a terminal electron acceptor and produces nitrous oxide. PMID- 16346788 TI - Comparison of methane production rate and coenzyme f(420) content of methanogenic consortia in anaerobic granular sludge. AB - The coenzyme F(420) content of granular sludge grown on various substrates and substrate combinations was measured, and the potential of the sludge to form methane (maximum specific methane production rate) from hydrogen, formate, acetate, propionate, and ethanol was determined. The F(420) content varied between 55 nmol g of volatile suspended solids (VSS) for sludge grown on acetate and 796 nmol g of VSS for sludge grown on propionate. The best correlation was found between the F(420) content and the potential activity for methane formation from formate; almost no correlation, however, was found with acetate as the test substrate. The ratio between the potential methanogenic activities (qch(4)) of sludges grown on various substrates and their F(420) content was in general highest for formate (48.2 mumol of CH(4) mumol of F(420) min) and lowest for propionate (6.9 mumol of CH(4) mumol of F(420) min) as test substrates. However, acetate-grown granular sludge with acetate as test substrate showed the highest ratio, namely, 229 mumol of CH(4) mumol of F(420) min. The data presented indicate that the F(420) content of methanogenic consortia can be misleading for the assessment of their potential acetoclastic methanogenic activity. PMID- 16346789 TI - General Biochemical Characterization of Thermostable Extracellular beta-Amylase from Clostridium thermosulfurogenes. AB - Clostridium thermosulfurogenes, an anaerobic bacterium which ferments starch into ethanol at 62 degrees C, produced an active extracellular amylase and contained intracellular glucoamylase but not pullulanase activity. The extracellular amylase was purified 2.4-fold, and its general physicochemical and catalytic properties were examined. The extracellular amylase was characterized as a beta amylase (1,4-alpha-d-glucan maltohydrolase) based on demonstration of exocleavage activity and the production of maltose with a beta-anomeric configuration from starch. The beta-amylase activity was stable and optimally active at 80 and 75 degrees C, respectively. The pH optimum for activity and the pH stability range was 5.5 to 6 and 3.5 to 6.5, respectively. The apparent [S](0.5V) and V(max) for beta-amylase activity on starch was 1 mg/ml and 60 U/mg of protein. Similar to described beta-amylase, the enzyme was inhibited by p-chloromercuribenzoate, Cu, and Hg; however, alpha- and beta-cyclodextrins were not competitive inhibitors. The beta-amylase was active and stable in the presence of air or 10% (vol/vol) ethanol. The beta-amylase and glucoamylase activities enabled the organism to actively ferment raw starch in the absence of significant pullulanase or alpha amylase activity. PMID- 16346790 TI - General Biochemical Characterization of Thermostable Pullulanase and Glucoamylase from Clostridium thermohydrosulfuricum. AB - Cell extracts of Clostridium thermohydrosulfuricum, an anaerobic bacterium which ferments starch into ethanol at 65 degrees C, contained both pullulanase and glucoamylase activities. The general physiochemical and catalytic properties of these enzyme activities were compared. Pullulanase and glucoamylase activities were stable and optimally active at 85 and 75 degrees C, respectively. The pH optima for activity and pH stability ranges were, respectively, 5.5 to 6 and 4.5 to 5.5 for pullulanase and 4 to 6 and 5 to 6 for glucoamylase. The apparent [S](0.5v) and V(max) for pullulanase activity on pullulan were 0.33 mg/ml and 2.6 U/mg of protein. The apparent [S](0.5v) and V(max) for glucoamylase activity on starch were of 0.41 mg/ml and 0.31 U/mg of protein. These enzymes were active and stable in the presence of air or 10% (vol/vol) ethanol. These enzyme activities allowed the organism to actively degrade raw starch into glucose in the absence of significant alpha-amylase activity. PMID- 16346791 TI - Simultaneous and Enhanced Production of Thermostable Amylases and Ethanol from Starch by Cocultures of Clostridium thermosulfurogenes and Clostridium thermohydrosulfuricum. AB - Clostridium thermohydrosulfuricum and Clostridium thermosulfurogenes produced ethanol and amylases with different components as primary metabolites of starch fermentation. Starch fermentation parameters were compared in mono- and cocultures of these two thermoanaerobes to show that the fermentation was dramatically improved as a consequence of coordinate action of amylolytic enzymes and synergistic metabolic interactions between the two species. Under given monoculture fermentation conditions, neither species completely degraded starch during the time course of the study, whereas in coculture, starch was completely degraded. In monoculture starch fermentation, C. thermohydrosulfuricum produced lower levels of pullulanase and glucoamylase, whereas C. thermosulfurogenes produced lower levels of beta-amylase and glucoamylase. In coculture fermentation, improvement of starch metabolism by each species was noted in terms of increased amounts and rates of increased starch consumption, amylase production, and ethanol formation. The single-step coculture fermentation completely degraded 2.5% starch in 30 h at 60 degrees C and produced 9 U of beta amylase per ml, 1.3 U of pullulanase per ml, 0.3 U of glucoamylase per ml, and >120 mM ethanol with a yield of 1.7 mol/mol of glucose in starch. The potential industrial applications of the coculture fermentation and the physiological basis for the interspecies metabolic interactions are discussed. PMID- 16346792 TI - Type 1 fimbria-mediated adhesion of enteric bacteria to grass roots. AB - Type 1 fimbriae of Klebsiella pneumoniae and Enterobacter agglomerans mediated bacterial adhesion to the roots of bluegrass, Poa pratensis. Purified, radiolabeled fimbriae bound to grass roots in vitro; binding was inhibited by alpha-methyl-d-mannoside or Fab fragments to the fimbriae. Anti-type 1 fimbriae Fab fragments and alpha-methyl-d-mannoside also inhibited adhesion of type 1 fimbriated bacteria to P. pratensis roots. It is proposed that associative nitrogen fixation by Klebsiella and Enterobacter strains also involves type 1 fimbriae, in addition to the type 3 fimbriae of Klebsiella spp. (T. K. Korhonen, E. Tarkka, H. Ranta, and K. Haahtela, J. Bacteriol. 155:860-865, 1983). PMID- 16346793 TI - In vitro adhesion of n(2)-fixing enteric bacteria to roots of grasses and cereals. AB - Nitrogen-fixing Klebsiella and Enterobacter strains isolated from several plants were assayed for fimbriae and for adhesion to plant roots in vitro. All eight Klebsiella strains formed type 3 fimbriae, and five strains also formed type 1 fimbriae; all 21 Enterobacter strains had type 1 fimbriae. Three strains of Klebsiella carrying either type 1, type 3, or no fimbriae were used as model organisms in developing an in vitro adhesion test. Adhesion was assayed with bacterial cells labeled with [H]leucine. Fifteen N(2)-fixing strains and the three model strains were compared for adhesion to the roots of seven grasses and five cereals. Type 3-fimbriated Klebsiella strains adhered better than the other strains, and type 3 fimbriae appeared to be major adhesins for the Klebsiella strains. Although variations between plants were observed, no host specificity for bacterial adhesion was found. PMID- 16346794 TI - Construction of killer wine yeast strain. AB - A double-stranded RNA plasmid which confers the superkiller phenotype was transferred into a wine yeast (Montrachet strain 522) and its leucine-requiring derivative (strain 694) by cytoduction, using the protoplast fusion technique. The killer wine yeast constructed completely suppressed the growth of killer sensitive strains of Saccharomyces cerevisiae in yeast extract-peptone-glucose medium at pH 4.5, whereas the killer effect was somewhat decreased at pH 3.5. The wine yeast harboring the killer factor also inhibited the growth of killer sensitive cells satisfactorily when it was grown in grape juice. PMID- 16346795 TI - Metabolism and Solubilization of Cellulose by Clostridium cellulolyticum H10. AB - When Clostridium cellulolyticum was grown with cellulose MN300 as the substrate, the rates of growth and metabolite production were found to be lower than those observed with soluble sugars as the substrate. At low cellulose concentrations, the growth yields were equal to those obtained with cellobiose. The main fermentation products from cellulose and soluble sugars were the same. Up to 15 mM of consumed hexose, a change in the metabolic pathway favoring lactate production similar to that observed with soluble sugars was found to occur concomitantly with a decrease in molar growth yield. With cellulose concentrations above 5 g/liter, accumulation of soluble sugars occurred once growth had ceased. Glucose accounted for 30% of these sugars. A kinetic analysis of cellulose solubilization revealed that cellulolysis by C. cellulolyticum involved three stages whatever cellulose concentration was used. Analysis of these kinetics showed three consecutive enzymatic activity levels having the same K(m) (0.8 g of cellulose per liter, i.e., 5 mM hexose equivalent) but decreasing values of V(max). The hypothesis is suggested that each step corresponds to differences in cellulose structure. PMID- 16346796 TI - Isolation and Characterization of a Cyclohexane-Metabolizing Xanthobacter sp. AB - An unusual Xanthobacter sp., capable of independent growth on cyclohexane as the sole source of carbon and energy, has been isolated from soil by using classical enrichment techniques. The mean generation time for growth on cyclohexane was 6 h. The microorganism showed a limited ability to utilize hydrocarbons, with only alicyclic hydrocarbons closely related to cyclohexane supporting growth. Ultrastructural studies indicated the presence of electron-transparent vesicles in the cyclohexane-grown Xanthobacter sp., but the presence of complex intracytoplasmic membranes could not be identified. A soluble inducible enzyme capable of oxidizing cyclohexane was identified in cell extracts. This enzyme had a pH optimum of 6.5, an absolute specificity for NADPH, and a stoichiometric requirement for molecular O(2) which was consistent with the formation of cyclohexanol. The enzyme showed no activity towards straight chain alkanes and only a limited activity towards unsaturated ring compounds. Enzymatic studies with cell extracts have indicated the main route of metabolism of cyclohexane by this Xanthobacter sp. to proceed via cyclohexane --> cyclohexanol --> cyclohexanone --> 1-oxa-2-oxocycloheptane (epsilon-caprolactone) --> 6 hydroxyhexanoate (6-hydroxycaproate) --> --> adipic acid. Alternative routes involving initial double hydroxylation of the cyclohexane ring may operate fortuituously but are unlikely to represent major pathways for the dissimilation of cyclohexane by this microorganism. PMID- 16346798 TI - A gram-negative anaerobic bacterium that utilizes o-methyl substituents of aromatic acids. AB - A novel strain of gram-negative anaerobic rods which utilized O-methyl substituents of monoaromatic acids as a sole organic source of carbon was isolated from municipal sewage sludge. Energy for growth seemed to be generated by an acetate formation pathway. The growth yield in defined medium was 7.9 g (dry weight) of cells per mol of ferulate utilized. This isolate and other O demethylating anaerobes may play a role in the turnover of acetate and the metabolism of highly methoxylated lignaceous materials in anaerobic environments. PMID- 16346799 TI - Effects of Nitrapyrin [2-Chloro-6-(Trichloromethyl) Pyridine] on the Obligate Methanotroph Methylosinus trichosporium OB3b. AB - [This corrects the article on p. 261 in vol. 47.]. PMID- 16346800 TI - Isotherm for Adsorption of Agrobacterium tumefaciens to Susceptible Potato (Solanum tuberosum L.) Tissues. AB - Potato tuber disks were submerged in suspensions containing 10 to 10 cells of Agrobacterium tumefaciens B6 per ml. After 60 min, the disks were rinsed and homogenized, and portions of the homogenates were plated to measure the number of adsorbed bacteria. At low initial bacterial concentrations (10/ml), 5 to 23% of the bacteria adsorbed. At higher bacterial concentrations, the corresponding value was approximately 1.2%. Adsorption was a reversible equilibrium process. Binding saturation was not achieved, and adsorbed bacteria were confined to monolayers on the surfaces of tissue prepared for scanning electron microscopy. Adsorption of strain B6 to potato tuber tissues is described accurately by the Freundlich adsorption isotherm and may be a nonspecific phenomenon. PMID- 16346801 TI - Study of Bacillus subtilis Endospores in Soil by Use of a Modified Endospore Stain. AB - The Schaeffer-Fulton endospore stain was modified so that it would stain Bacillus subtilis endospores in soil smears. The modified stain differentiated among dormant spores, spores undergoing activation, and spores which had germinated but had not yet shown outgrowth. These differentiations were seen for spores in soil and for pure spore preparations in the laboratory. This stain was used to show reversible B. subtilis spore activation promoted by an Ensifer adhaerens-like indigenous bacterium in soil and by pure cultures of E. adhaerens added to spores in the laboratory. Under the specific conditions in the laboratory, spore germination did not proceed beyond the activation stage, and relatively little change occurred in the numbers of both E. adhaerens and B. subtilis. This was also true in soil, although some germination with destruction of spores and vegetative cells did occur if the soil had been nutritionally enriched by preincubation with incorporated ground alfalfa. PMID- 16346802 TI - Characterization of bacteria by particle beam mass spectrometry. AB - A technique is described for detecting and characterizing bacteria on a single particle basis by mass spectrometry. The method involves generation of a particle beam of single whole cells which are rapidly volatilized and ionized in vacuum in the ion source of a quadrupole mass spectrometer. The particle beam can be generated, with minimal sample handling, from a naturally occurring aerosol or from a solution of bacteria that can be dispersed as an aerosol. The mass spectrum is generated by successively measuring the average intensities of different mass peaks. The average intensity is obtained by measuring the ion intensity distribution at the particular mass (m/e) for ion pulses from more than 1,000 bacteria particles. Bacillus cereus, Bacillus subtilis, and Pseudomonas putida samples were analyzed to test the capability of the instrument for differentiating among species of bacteria. Significant ion-intensity information was produced over the m/e range of 50 to 300, an improvement over previous pyrolysis-mass spectrometry results. The complex mass spectra contained a few unique peaks which could be used for the differentiation of the bacteria. A statistical analysis of the variations in peak intensities among the three bacteria provided a quantitative measure of the reproducibility of the instrument and its ability to differentiate among bacteria. The technique could lead to a new rapid method for the analysis of microorganisms and could be used for the detection of airborne pathogens on a continuous, real-time basis. PMID- 16346803 TI - Effect of Sym Plasmid Curing on Symbiotic Effectiveness in Rhizobium fredii. AB - A mutant, USDA 206C, of Rhizobium fredii USDA 206 was obtained by passage on acridine plates. This mutant was cured of its 197-megadalton Sym plasmid but retained its symbiotic effectiveness. Multiple plasmid and chromosomally borne nif gene copies have previously been shown in R. fredii USDA 206. HindIII and EcoRI restriction enzyme digests of plasmid and total DNA showed that at least two nif gene copies are probably missing in USDA 206C. To compare the symbiotic effectiveness of USDA 206 and USDA 206C, plant tests were carried out. Statistically significant differences were obtained for nodule number, nodule mass, nitrogenase activity per plant, nitrogenase specific activity, and total plant dry weight. There was an apparent correlation between loss of Sym plasmidborne nif gene copies and reduction of overall symbiotic effectiveness. Delayed nodulation by strain USDA 206C relative to strain USDA 206 also indicated an association with the loss of plasmidborne nodulation functions and the reduced symbiotic effectiveness of strain USDA 206C. PMID- 16346804 TI - Enhancement of Pyrroloquinoline Quinone Production and Polyvinyl Alcohol Degradation in Mixed Continuous Cultures of Pseudomonas putida VM15A and Pseudomonas sp. Strain VM15C with Mixed Carbon Sources. AB - In a mixed continuous culture of Pseudomonas putida VM15A and Pseudomonas sp. strain VM15C with polyvinyl alcohol (PVA) as the sole source of carbon, growth of the PVA-degrading bacterium VM15C and, hence, PVA degradation were limited by the growth factor, pyrroloquinoline quinone, produced by VM15A. Feeding of a carbon source for VM15A, ethanol, with PVA enhanced pyrroloquinoline quinone production and caused increases in the VM15C population and PVA degradation in a mixed continuous culture. There was an optimum range for PVA degradation of the ethanol concentration, although pyrroloquinoline quinone concentrations in continuous mixed cultures increased with increasing ethanol concentration. PMID- 16346805 TI - Mechanism of the Heat Sensitization of Bacillus subtilis Spores by Ethidium Bromide. AB - Pretreatment with ethidium bromide (5 mug/ml) followed by a water wash had no effect on unheated Bacillus subtilis spores, but the viability of these spores after heating was much lower than that of similarly heated spores exposed to water alone. The fate of water- or ethidium bromide-treated spores, unheated or heated, was followed by allowing them to germinate and outgrow in a minimal or a complex liquid medium. Spores exposed to ethidium bromide and then heated (85 degrees C, 10 min) exhibited a developmental block during germination and outgrowth. Many of them were blocked at the stage when the bacterium emerged from the germinated spore. When 0.35 mug of ethidium bromide per ml was added to heated spores in the germination-growth medium, the outgrowth of heated spores was inhibited to the same extent as were pretreated spores. Ethidium bromide acted in the first hour of germination of heated spores since addition after this time was ineffective in inhibiting recovery events. Repair of heat-damaged spore DNA was detected during the first 2 h of germination. The addition of ethidium bromide (final concentration, 0.35 mug/ml) inhibited DNA repair during early outgrowth. Increased sensitivity of spores to heat after pretreatment with sublethal concentrations of ethidium bromide was due to the inhibition of the repair of heat-damaged DNA. PMID- 16346806 TI - Role of a Ferric Ion-Reducing System in Sulfur Oxidation of Thiobacillus ferrooxidans. AB - The properties of a ferric ion-reducing system which catalyzes the reduction of ferric ion with elemental sulfur was investigated with a pure strain of Thiobacillus ferrooxidans. In anaerobic conditions, washed intact cells of the strain reduced 6 mol of Fe with 1 mol of elemental sulfur to give 6 mol of Fe, 1 mol of sulfate, and a small amount of sulfite. In aerobic conditions, the 6 mol of Fe produced was immediately reoxidized by the iron oxidase of the cell, with a consumption of 1.5 mol of oxygen. As a result, Fe production was never observed under aerobic conditions. However, in the presence of 5 mM cyanide, which completely inhibits the iron oxidase of the cell, an amount of Fe production comparable to that formed under anaerobic conditions was observed under aerobic conditions. The ferric ion-reducing system had a pH optimum between 2.0 and 3.8, and the activity was completely destroyed by 10 min of incubation at 60 degrees C. A short treatment of the strain with 0.5% phenol completely destroyed the ferric ion-reducing system of the cell. However, this treatment did not affect the iron oxidase of the cell. Since a concomitant complete loss of the activity of sulfur oxidation by molecular oxygen was observed in 0.5% phenol-treated cells, it was concluded that the ferric ion-reducing system plays an important role in the sulfur oxidation activity of this strain, and a new sulfur-oxidizing route is proposed for T. ferrooxidans. PMID- 16346807 TI - Dissimilation of Carbon Monoxide to Acetic Acid by Glucose-Limited Cultures of Clostridium thermoaceticum. AB - Clostridium thermoaceticum was cultivated in glucose-limited media, and the dissimilation of CO to acetic acid was evaluated. We found that cultures catalyzed the rapid dissimilation of CO to acetic acid and CO(2), with the stoichiometry obtained for conversion approximating that predicted from the following reaction: 4CO + 2H(2)O --> CH(3)CO(2)H + 2CO(2). Growing cultures formed approximately 50 mmol (3 g) of CO-derived acetic acid per liter of culture, with the rate of maximal consumption approximating 9.1 mmol of CO consumed/h per liter of culture. In contrast, resting cells were found not to dissimilate CO to acetic acid. CO was incorporated, with equal distribution between the carboxyl and methyl carbons of acetic acid when the initial cultivation gas phase was 100% CO, whereas CO(2) preferentially entered the carboxyl carbon when the initial gas phase was 100% CO(2). Significantly, in the presence of saturating levels of CO, CO(2) preferentially entered the methyl carbon, whereas saturating levels of CO(2) yielded CO-derived labeling predominantly in the carboxyl carbon. These findings are discussed in relation to the path of carbon flow to acetic acid. PMID- 16346808 TI - Effect of Cultural Practices, Soil Phosphorus, Potassium, and pH on the Incidence of Fusarium Head Blight and Deoxynivalenol Levels in Wheat. AB - In a survey conducted in 1984 in Essex, Lambton, and Middlesex Counties of Ontario, Canada, greater incidences of head blight and greater concentrations of deoxynivalenol in grain were observed in fields of winter wheat planted after corn than in those planted after soybeans, barley, and mixed grains. Neither head blight nor deoxynivalenol level were correlated significantly with soil P, K, or pH. Head blight was reduced in wheat planted after corn where the residues from a preceding corn crop were plowed down or where seed was treated with Vitaflo 280. However, head blight and deoxynivalenol levels were not affected significantly by the level of N application, cultivar, other diseases or herbicides. PMID- 16346809 TI - Conserved Nodulation Genes in Rhizobium meliloti and Rhizobium trifolii. AB - Plasmids which contained wild-type or mutated Rhizobium meliloti nodulation (nod) genes were introduced into NodR. trifolii mutants ANU453 and ANU851 and tested for their ability to nodulate clover. Cloned wild-type and mutated R. meliloti nod gene segments restored ANU851 to Nod, with the exception of nodD mutants. Similarly, wild-type and mutant R. meliloti nod genes complemented ANU453 to Nod, except for nodCII mutants. Thus, ANU851 identifies the equivalent of the R. meliloti nodD genes, and ANU453 specifies the equivalent of the R. meliloti nodCII genes. In addition, cloned wild-type R. trifolii nod genes were introduced into seven R. meliloti Nod mutants. All seven mutants were restored to Nod on alfalfa. Our results indicate that these genes represent common nodulation functions and argue for an allelic relationship between nod genes in R. meliloti and R. trifolii. PMID- 16346810 TI - Transpositional Mutagenesis of Thiobacillus novellus and Thiobacillus versutus. AB - Transpositional mutagenesis of Thiobacillus novellus by Tn501 was achieved by means of the incompatibility of IncP plasmids. Tn501 insertion caused three types of mutant phenotypes: isoleucine auxotrophy, lysine auxotrophy, and a reduced ability to oxidize reduced sulfur compounds and to fix CO(2). Oxidation rates for elemental sulfur (S), thiosulfate (S(2)O(3)), and tetrathionate (S(4)O(6)) in mutants of the latter type were reduced relative to those of the nonmutant control strain. Incorporation of labeled bicarbonate (HCO(3)) was also significantly impaired. Although suicide vehicles were not useful for the introduction of transposons into T. novellus, this method was effective for the Tn1721-induced mutagenesis of Thiobacillus versutus. Tn1721 insertions resulted in the loss of the natural resistance of T. versutus to arsenate and gentamicin and in auxotrophies for isoleucine-valine, arginine, phenylalanine, valine, and panthothenate. Transpositional mutagenesis by either method should prove to be a useful tool for further study of these and other members of the genus Thiobacillus. PMID- 16346811 TI - Rate of bacterial mortality in aquatic environments. AB - A method is proposed which provides a minimum estimate of the rate of bacterial mortality in growing natural populations of planktonic bacteria. This estimate is given by the rate of decrease of radioactivity from the DNA of a [H]thymidine labeled natural assemblage of bacteria after all added thymidine has been exhausted from the medium. Results obtained from river water, estuarine water, and seawater show overall bacterial mortality rates in the range 0.010 to 0.030 h, in good agreement with the range of growth rates measured in the same environments. Use of selective filtration through Nuclepore filters (pore size, 2 mum) allowed us to determine the contribution of microzooplankton grazing to overall bacterial mortality. Grazing rates estimated by this method ranged from 0 to 0.02 h. PMID- 16346812 TI - Isolation and Some Properties of the Enzyme That Transforms Eremofortin C to PR Toxin. AB - PR toxin and eremofortin C are secondary metabolites of Penicillium roqueforti. The chemical structures of these two compounds are closely related to each other and differ only by an aldehyde and an alcohol group at the C-12 position. In an effort to better understand the biosynthesis of PR toxin, we discovered the enzyme of P. roqueforti that is responsible for the transformation of eremofortin C to PR toxin. The maximum activity of the enzyme in the culture medium was found to occur on day 13, which corresponded to the maximal production of PR toxin in the medium. The enzyme was isolated and purified from the culture medium and the mycelium of the fungus, respectively, through a procedure involving ammonium sulfate fractionation and DEAE-cellulose chromatography. The specific activity increased 20- and 8-fold, respectively, and the yield was 33.3 and 21.6%, respectively, for the enzyme from the medium and mycelium. The optimal pH for the enzyme reaction was ca. pH 5.6. The enzyme reaction was temperature dependent. The rates followed a linear time course when it catalyzed the transformation at 30 degrees C and decayed with time when reacted at higher temperatures. At 100 degrees C, the enzyme activity was completely lost. The K(m) and V(max) of the enzyme as determined at 30 degrees C were 0.02 mM and 4.0 mumol/min per mg, respectively. The molecular weight of the enzyme was estimated by gel filtration on a high-pressure liquid chromatography I-250 protein column to be ca. 40,000. PMID- 16346813 TI - Isolation of a Butyrate-Utilizing Bacterium in Coculture with Methanobacterium thermoautotrophicum from a Thermophilic Digester. AB - Sludge from a thermophilic, 55 degrees C digester produced methane without a lag period when enriched with butyrate. The sludge was found by most-probable-number enumeration to have ca. 5 x 10 butyrate-utilizing bacteria per ml. A thermophilic butyrate-utilizing bacterium was isolated in coculture with Methanobacterium thermoautotrophicum. This bacterium was a gram-negative, slightly curved rod, occurred singly, was nonmotile, and did not appear to produce spores. When this coculture was incubated with Methanospirillum hungatei at 37 degrees C, the quantity of methane produced was less than 5% of the methane produced when the coculture was incubated at 55 degrees C, the routine incubation temperature. The coculture required clarified digester fluid. The addition of yeast extract to medium containing 5% clarified digester fluid stimulated methane production when a Methanosarcina sp. was present. Hydrogen in the gas phase prevented butyrate utilization. However, when the hydrogen was removed, butyrate utilization began. Penicillin G and d-cycloserine caused the complete inhibition of butyrate utilization by the coculture. The ability of various ecosystems to convert butyrate to methane was studied. Marine sediments enriched with butyrate required a 2-week incubation period before methanogenesis began. Hypersaline sediments did not produce methane after 3 months when enriched with butyrate. PMID- 16346814 TI - Methanogenesis in an Upflow Anaerobic Sludge Blanket Reactor at pH 6 on an Acetate-Propionate Mixture. AB - High-rate anaerobic digestion can be applied in upflow anaerobic sludge blanket reactors for the treatment of various wastewaters. In upflow anaerobic sludge blanket reactors, sludge retention time is increased by a natural immobilization mechanism (viz. the formation of a granular type of sludge). When this sludge is cultivated on acid-containing wastewater, the granules mainly consist of an acetoclastic methanogen resembling Methanothrix soehngenii. This organism grows either in rods or in long filaments. Attempts to cultivate a stable sludge consisting predominantly of Methanosarcina sp. on an acetate-propionate mixture as substrate by lowering the pH from 7.5 during the start-up to approximately 6 failed. After 140 days of continuous operation of the reactor a filamentous organism resembling Methanothrix soehngenii prevailed in the sludge. The specific methanogenic activity of this sludge on acetate-propionate was optimal at pH 6.6 to 6.8 and 7.0 to 7.2, respectively. PMID- 16346815 TI - Selective Isolation of Bacillus sphaericus from Soil by Use of Acetate as the Only Major Source of Carbon. AB - A simple chemically defined medium containing sodium acetate (37 or 74 mM) as the only major source of carbon was inoculated with soil from various locations. The bacteria which grew most rapidly at 30 degrees C were almost entirely strains of Bacillus sphaericus and Arthrobacter species. Pasteurization of the soils made the medium selective for B. sphaericus. Several new strains were isolated, and the peptidoglycans of their cell walls were examined. PMID- 16346816 TI - Separation and purification of bacteria from soil. AB - Bacteria were released and separated from soil by a simple blending centrifugation procedure. The percent yield of bacterial cells (microscopic counts) in the supernatants varied over a wide range depending on the soil type. The superantants contained large amounts of noncellular organic material and clay particles. Further purification of the bacterial cells was obtained by centrifugation in density gradients, whereby the clay particles and part of the organic materials sedimented. A large proportion of the bacteria also sedimented through the density gradient, showing that they had a buoyant density above 1.2 g/ml. Attachment to clay minerals and humic material may account for this apparently high buoyant density. The percent yield of cells was negatively correlated with the clay content of the soils, whereas the purity was positively correlated with it. The cell size distribution and the relative frequency of colony-forming cells were similar in the soil homogenate, the supernatants after blending-centrifugation, and the purified bacterial fraction. In purified bacterial fraction from a clay loam, the microscopically measured biomass could account for 20 to 25% of the total C and 30 to 40% of the total N as cellular C and N. The amount of cellular C and N may be higher, however, owing to an underestimation of the cell diameter during fluorescence. A part of the contamination could be ascribed to extracellular structures as well as partly decayed cells, which were not revealed by fluorescence microscopy. PMID- 16346817 TI - Bacterial biovolume and biomass estimations. AB - The biomass of bacterial populations in aquatic ecosystems is often estimated by measuring bacterial biovolume and converting this into biomass in terms of carbon. A reliable conversion factor relating the measured bacterial biovolume to bacterial carbon content is essential for this approach. Based on direct measurements of bacterial cell carbon content, cell number, and biovolume, I have derived an average conversion factor of 5.6 x 10 g of C mum. This conversion factor is 3.4 to 6.6 times higher than most theoretically derived factors currently in use. Both bacterial biomass and bacterial production in aquatic ecosystems may thus have been seriously underestimated. PMID- 16346818 TI - Growth of Agaricus campestris NRRL 2334 in the Form of Pellets. AB - The production of pellets of the fungus Agaricus campestris NRRL 2334 was studied in submerged fermentation with peat extract as the main substrate source. Pellets up to 6 mm in diameter were obtained when the peat extract was diluted to reduce the concentration of growth inhibitors. Yeast extract and yeast extract plus glucose were the most effective nutrient supplements in the diluted peat extract media and stimulated the formation of large pellets which contained 44.4% crude protein, 2.8% fat, and 9% ash (dry weight basis). No solid supports were required for the growth of the pellets. The effects on the growth morphology of several dilution ratios of the peat extract, rates of agitation and aeration, and time were investigated. PMID- 16346820 TI - Minimum threshold for hydrogen metabolism in methanogenic bacteria. AB - Methanogenic isolates did not consume hydrogen below partial pressures of 6.5 Pa. Thus, in contrast to a previous report, results from pure-culture studies do not invalidate the threshold model for methane production from hydrogen in sediments. PMID- 16346819 TI - Association of Rhizobium Strains with Roots of Trifolium repens. AB - TWO TECHNIQUES WERE USED TO ASSESS THE BINDING OF RHIZOBIA TO CLOVER ROOTS: indirect counting after radiolabeling the bacteria and direct counting by using phase-contrast microscopy. Microscopic observations revealed a large variability in the number of bacteria associated with individual root hairs. This variability made unbiased counting by microscopy difficult. Systematic examination of all visible root hairs and "blind" counting of coded strains and treatments were adopted to minimize observer bias. The validity of the radiolabeling method was also examined in some detail. The reproducibility of results from this method was satisfactory. However, drawbacks of this method included its lack of sensitivity and its failure to distinguish between bacteria attached to mature root hairs, emerging root hairs, and undifferentiated epidermal cells. The method also failed to distinguish between individual bacteria and any aggregates that may be present. The ability of a number of chosen mutant strains of Rhizobium trifolii and their corresponding parent strains, as well as a number of nonhomologous strains, to bind to clover roots was assessed by using both of these methods. Our results gave no indication of specificity of R. trifolii binding to clover roots. 2-Deoxy-d-glucose did not appear to have a major inhibitory effect on the attachment of rhizobia to the host root, which suggests that lectin cross bridging is not an obligatory step in the initiation of infection even though it may occur under some conditions. The presence or absence of the symbiotic plasmid was not correlated with bacterial adherence to the host plant root. Since host specificity functions are carried on this plasmid, our results suggest that binding of rhizobia to the legume root is not the basis of host specificity. PMID- 16346821 TI - Selective Medium for Mosquito-Pathogenic Strains of Bacillus sphaericus. AB - A selective medium (BATS), which contains arginine as the sole carbon and nitrogen source and which also contains streptomycin, allowed growth of 18 mosquito-pathogenic strains of Bacillus sphaericus. It inhibited the growth of 68% of the nonpathogenic B. sphaericus strains tested as well as other Bacillus spp. and aquatic bacteria. PMID- 16346822 TI - Effect of Photon Fluence Rate and Specific Growth Rate on Geosmin Production of the Cyanobacterium Oscillatoria brevis (Kutz.) Gom. AB - The production of geosmin from the cyanobacterium Oscillatoria brevis was studied as a function of the photon fluence rate and appears to be related to the chlorophyll content. PMID- 16346823 TI - Modified medium for the oxidation-fermentation test in the identification of marine bacteria. AB - Using the medium of ZoBell as a basal formulation, we developed a medium to determine the oxidative-fermentative metabolism in marine bacteria. Different oxidative and fermentative collection strains, as well as a group of unidentified marine bacteria, were tested on modified ZoBell medium and other conventional media. In general, superior results were obtained with the modified ZoBell medium since it supported good bacterial growth and gave more clear-cut oxidative reactions. PMID- 16346824 TI - Genetic Rearrangement of Plasmids: In Vivo Recombination between a Dehalogenation Plasmid and Multiple-Resistance Plasmid RP4 in Pseudomonas sp. AB - When Moraxella plasmid pUO1 encoding haloacetate dehalogenase and mercury resistance coexisted with IncP-1 plasmid RP4 in Pseudomonas sp., genetic exchange between the plasmids often occurred, probably by site-specific recombination. The recombinant plasmids obtained were classified into four groups on the basis of phenotype. Representative plasmids for each group were analyzed for DNA composition and function, and the mechanism for the formation of these plasmids was sought. They were inherited stably in Escherichia coli and a Pseudomonas sp. PMID- 16346825 TI - High Production of Thermostable beta-Galactosidase of Bacillus stearothermophilus in Bacillus subtilis. AB - By cloning the beta-galactosidase gene of Bacillus stearothermophilus IAM11001 (ATCC 8005) into Bacillus subtilis, enzyme production was enhanced 50 times. beta Galactosidase could be purified to 80% homogeneity by incubating the cell extract of B. subtilis at 70 degrees C for 15 min, followed by centrifugation to remove the denatured proteins. Because of its heat stability and ease of production, beta-galactosidase is suitable for application in industrial processes. PMID- 16346826 TI - Improvement of protoplast regeneration by growing yeast cells hypertonically. AB - The high osmotic pressure provided for in vitro transformation processes to support naked protoplasts of Saccharomyces cerevisiae was found to hurt regeneration of the protoplasts to walled cells. Preadaptation of the cells by growth under the same hypertonic conditions as those used for transformation doubled the rate of protoplast regeneration and in turn resulted in a higher yield of transformants. PMID- 16346827 TI - Degradation of Halogenated Aliphatic Compounds by Xanthobacter autotrophicus GJ10. AB - [This corrects the article on p. 674 in vol. 49.]. PMID- 16346828 TI - Only Part of the Protoxin Gene of Bacillus thuringiensis subsp. berliner 1715 Is Necessary for Insecticidal Activity. AB - [This corrects the article on p. 708 in vol. 49.]. PMID- 16346829 TI - Survival of bacteria and fungi in relation to water activity and the solvent properties of water in biopolymer gels. AB - Survival of bacteria (Rhizobium, Agrobacterium, and Arthrobacter spp.), fungal spores (Penicillium sp.), and yeasts (Saccharomyces sp.) was studied in relation to water activity (a(w)) and the presence of nutritive solutes. The cells were entrapped in polysaccharide gels, as is done to immobilize cells or enzymes, and then dehydrated. The number of living cells (10 g of dry polymer) remained constant for periods of storage of >3 years at 28 degrees C when the inocula were kept at an a(w) of <0.069. At a(w) values between 0.069 and 0.83 the number of survivors diminished more and more rapidly as the a(w) was raised. For a given a(w) and organism, there were large differences in survival rate as a function of the nutritive solutes used to culture the microorganisms. Low-molecular-weight compounds (with three or five carbon atoms) had a deleterious effect on survival, whereas compounds of higher molecular weight (C(6) to C(12)) had a protecting effect. Thus, the a(w) alone was not a sufficient explanation for the deterioration of the inocula. Survival seemed to be more directly related to some properties of the water in the biopolymer. New concepts such as the discontinuity of properties of water and the point of mobilization of solutes, already proposed by Duckworth and Kelly (J. Food Technol. 8:105-113, 1973) and Seow (J. Sci. Food Agric., 26:535-536, 1975), have been taken into consideration to explain the interactions of water with the biopolymer and their specific effects on the microorganisms. PMID- 16346830 TI - Effect of reactor turbulence on the binding-protein-mediated aspartate transport system in thin wastewater biofilms. AB - This research documents an effect of reactor turbulence on the ability of gram negative wastewater biofilm bacteria to actively transport l-aspartate via a binding-protein-mediated transport system. Biofilms which were not preadapted to turbulence and which possessed two separate and distinct aspartate transport systems (systems 1 and 2) were subjected to a turbulent flow condition in a hydrodynamically defined closed-loop reactor system. A shear stress treatment of 3.1 N . m for 10 min at a turbulent Reynolds number (Re = 11,297) inactivated the low-affinity, high-capacity binding-protein-mediated transport system (system 2) and resolved the high-affinity, low-capacity membrane-bound proton symport system (system 1). The K(t) and V(max) values for the resolved system were statistically similar to K(t) and V(max) values for system 1 when system 2 was inactivated either by osmotic shock or arsenate, two treatments which are known to inactivate binding-protein-mediated transport systems. We hypothesize that shear stress disrupts system 2 by deforming the outer membranes of the firmly adhered gram negative bacteria. PMID- 16346831 TI - Biological Delignification of Aspen Wood by Solid-State Fermentation with the White-Rot Fungus Merulius tremellosus. AB - Solid-state fermentation of aspen (Populus tremuloides) wood with Merulius tremellosus for 8 weeks removed 52% of the lignin but only 12% of the total wood weight, and increased the cellulase digestibility to 53% from 18%. Water-soluble and enzyme-solubilized lignin degradation products accumulated. Delignification was fastest at temperatures between 25 and 32.5 degrees C and at a water-to-wood ratio of 2. Initial pH values between 4 and 6 were optimal; M. tremellosus acidified the wood to below pH 3.5 as it grew. The fungus tolerated CO(2) concentrations of at least 14% and O(2) concentrations down to 7% in the bulk gas phase. Both simple and complex nitrogen supplements inhibited delignification. Supplementary KH(2)PO(4), MgSO(4), CaCl(2), thiamine, and trace elements had little effect on the fermentation. Four isolates of M. tremellosus had very similar abilities to delignify aspen wood. Biological delignification with M. tremellosus may be a useful pretreatment for enzymatic saccharification or ruminant feeding. PMID- 16346832 TI - Isolation and characterization of a moderately halophilic methanogen from a solar saltern. AB - A moderately halophilic methanogenic bacterium was enriched with trimethylamine and isolated from the sediment of a solar salt pond (total dissolved solids of pond water, 250 g/liter; pH 7.5). The isolate (strain SF1, DSM 3243) was an irregular coccus which stained gram negative, with a diameter of 1 mum and a thin monolayered cell wall. The organism grew singly, in pairs, and in irregular clumps. Colonies were tannish yellow, circular, with entire edges, and about 1 mm in diameter within 1 week. Only methylamines or methanol was used for growth and methanogenesis. Most rapid growth (doubling time, 10.2 h) occurred at a temperature of 37 degrees C and a pH of 7.4. The optimum NaCl concentration was 2.1 M. Yeast extract or rumen fluid was required. The isolate was lysed by sodium dodecyl sulfate (0.1 g/liter) and was sensitive to chloramphenicol. The G+C content of the DNA was 41 (+/-1) mol%. PMID- 16346833 TI - Regulation of beta-1, 4-Glucosidase Expression by Candida wickerhamii. AB - Candida wickerhamii NRRL Y-2563 expressed beta-glucosidase activity (3 to 8 U/ml) constitutively when grown aerobically in complex medium containing either glycerol, succinate, xylose, galactose, or cellobiose as the carbon source. The addition of a high concentration of glucose (>75 g/liter) repressed beta glucosidase expression (<0.3 U/ml); however, this yeast did produce beta glucosidase when the initial glucose concentration was 50 mum in size) were measured during a 19-day period in eutrophic Frederiksborg Slotsso, Denmark, as well as in experimental enclosures (containing 5.3 m of lake water). The fish and nutrients of the enclosures were manipulated. In enclosures without fish, large increases in ingestion by zooplankton >140 mum in size were found (up to 3 mug of C liter h), compared with values less than 0.3 mug of C liter h in the enclosures with fish and in the open lake. Daphnia cucullata and D. galeata dominated the community of zooplankton of >140 mum. Ingestion rates for zooplankton between 50 and 140 mum decreased after a period of about 8 days, in all enclosures and in the lake, to values below 0.1 mug of C liter h. On the last 2 sampling days, somewhat higher values were observed in the enclosures with fish present. The >50-mum zooplankton ingested 48 to 51% of the bacterial net secondary production in enclosures without fish, compared to 4% in the enclosures with added fish. Considering the sum of bacterial secondary production plus biomass change, 35 to 41% of the available bacteria were ingested by zooplankton of >50 mum in the enclosures without fish, compared with 4 to 6% in the enclosures with added fish and 21% in the open lake. Fish predation reduced the occurrence of zookplankton sized >50 mum and thus left a large proportion of the available bacteria to zooplankton sized <50 mum. In fact, there were 4.6 x 10 to 5.0 x 10 flagellates (4 to 8 mum in size) ml in the enclosures with fish added as well as in the lake, compared with 0.5 x 10 to 2.3 x 10 ml in the enclosures without fish. This link in the food chain was reduced when fish predation on zooplankton was eliminated and a direct route of dissolved organic matter, via the bacteria to the zooplankton, was established. PMID- 16346845 TI - Marine ammonia- and nitrite-oxidizing bacteria: serological diversity determined by immunofluorescence in culture and in the environment. AB - Immunofluorescence assays for marine ammonium- and nitrite-oxidizing bacteria were used to assess the diversity of nitrifying bacteria isolated from marine environments. The antisera show relatively broad specificity, in that each reacts with several strains of the same physiological type as the strain to which the antiserum was prepared. The antisera do not, however, react with any strains of differing physiological type. Seventy percent of the 30 unidentified ammonium oxidizing isolates tested reacted with one or both of the antisera produced to marine ammonium-oxidizing strains, and 8 of the 9 unidentified nitrite-oxidizing strains tested reacted with 1 or more of the 3 nitrite oxidizer antisera used. Ammonium- and nitrite-oxidizing bacteria were enumerated in samples taken in a depth profile (to 750 m) in the Southern California Bight by immunofluorescence assays for two ammonium oxidizers and two nitrite oxidizers. Average abundances of the two types of nitrifiers were 3.5 x 10 and 2.8 x 10 cells liter, respectively. Nitrifiers constitute 0.1 to 0.8% of the total bacterial population in these samples. PMID- 16346846 TI - Rates of Drying and Survival of Rhizobium meliloti Strains During Storage at Different Relative Humidities. AB - An investigation was made of the survival of six strains of Rhizobium meliloti filtered on membrane filters and held in atmospheres of controlled relative humidities (RH) of from 0 to 100% at 30 degrees C in the presence of air. The rate of water loss in the desiccator was determined by the humidity-controlling solution used. Drying was accelerated by a mild evacuation of the desiccator during the drying step. Survival rates of R. meliloti strains were much higher after slow drying to 0% RH than immediately after rapid drying. Fast drying (drying period less than 3.4 h) was shown to adversely affect the tolerance to storage at all RH values tested (no survival after 2 to 5 days of storage). When survival during storage was measurable (after slow drying), the optimum RH values for storage were 43% for strains A145 and Wu498, 22 to 43% for strains RCR2011, Wu499, and Ar16, and 83% for strain RCR2004. The most favorable drying periods were 8, 9.2, 14.2, and 50.1 h for the subsequent storage of strain RCR2011 at RH values of 0, 22, 43, and 83%, respectively. The damaging effects of rapid drying on the tolerance of strain RCR2011 to storage at different RH values could be prevented either by rehydration and subsequent slow redrying or incomplete rapid drying followed by slow drying. It is suggested that R. meliloti strains are susceptible to desiccation stresses. However, the quantitative differences among strains appear to be large enough to permit selection with regard to tolerance to desiccation and storage in dried states. PMID- 16346847 TI - Cellulolytic Activity of Clostridium acetobutylicum. AB - Clostridium acetobutylicum NRRL B527 and ATCC 824 exhibited extracellular and cell-bound endoglucanase and cellobiase activities during growth in a chemically defined medium with cellobiose as the sole source of carbohydrate. For both strains, the endoglucanase was found to be mainly extracellular (70 to 90%) during growth in continuous or batch cultures with the pH maintained at 5.2, whereas the cellobiase was mainly cell associated (60 to 90%). During continuous cultivation of strain B527 with cellobiose as the limiting nutrient, maximum production of the endoglucanase and cellobiase occurred at pH values of 5.2 and 4.8, respectively. In the carbon-limited continuous cultures, strain 824 produced similar levels of endoglucanase, cellobiosidase, and cellobiase activities regardless of the carbon source used. However, in ammonium- or phosphate-limited cultures, with an excess of glucose, only 1/10 of the endoglucanase was produced, and neither cellobiosidase nor cellobiase activities were detectable. A crude extracellular enzyme preparation from strain B527 hydrolyzed carboxymethylcellulose and phosphoric acid-swollen cellulose readily and microcrystalline cellulose (A vicel) to a lesser extent. Glucose accounted for more than 90% of the reducing sugar produced by the hydrolysis of acid-swollen cellulose and Avicel. Strain B527 did not grow in medium with acid-swollen cellulose as the sole source of carbohydrate, although it grew readily on the products obtained by hydrolyzing the cellulose in vitro with a preparation of extracellular cellulase derived from the same organism. PMID- 16346848 TI - Muramic Acid measurements for bacterial investigations in marine environments by high-pressure liquid chromatography. AB - Muramic acid, a constituent of procaryotic cell walls, was assayed by high pressure liquid chromatography in samples from several marine environments (water column, surface microlayer, and sediment) and a bacterial culture. It is used as a microbial biomass indicator. The method gave a good separation of muramic acid from interfering compounds with satisfactory reproducibility. A pseudomonad culture had a muramic acid content of 4.7 x 10 to 5.3 x 10 mug per cell during growth. In natural water samples, highly significant relationships were found between muramic acid concentrations and bacterial numbers for populations of 10 to 10 cells per liter. The muramic acid content in natural marine water decreased from 5.3 x 10 to 1.6 x 10 mug per cell with increasing depth. In coastal sediments exposed to sewage pollution, concentrations of muramic acid, ATP, organic carbon, and total amino acids displayed a parallel decrease with increasing distance from the sewage outlet. Advantages of muramic acid measurement by high-pressure liquid chromatography are its high sensitivity and reduction of preparation steps, allowing a short time analysis. PMID- 16346849 TI - Studies on Cellulose Hydrolysis by Acetivibrio cellulolyticus. AB - Acetivibrio cellulolyticus extracellular cellulase extensively hydrolyzed crystalline celluloses such as Avicel (FMC Corp., Food and Pharmaceutical Products Div., Philadelphia, Pa.) but only if it was desalted and supplemented with Ca. The Ca effect was one of increased enzyme stability in the presence of the ion. Although preincubation of the cellulase complex at 40 degrees C for 5 h without added Ca had a negligible effect on endoglucanase activity or on the subseqent hydrolysis of amorphous cellulose, the capacity of the enzyme to hydrolyze crystalline cellulose was almost completely lost. Adsorption studies showed that 90% of the Avicel-solubilizing component of the total enzyme preparation bound to 2% Avicel at 40 degrees C. Under these conditions, only 15% of the endoglucanase and 25% of the protein present in the enzyme preparation adsorbed to the substrate. The protein profile of the bound enzyme, as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was complex and distinctly different from the profile observed for total cellulase preparations. The specific activity of A. cellulolyticus cellulase with respect to Avicel hydrolysis was compared with that of commercially available Trichoderma reesei cellulase. PMID- 16346850 TI - Comparative Amperometric Study of Uptake Hydrogenase and Hydrogen Photoproduction Activities between Heterocystous Cyanobacterium Anabaena cylindrica B629 and Nonheterocystous Cyanobacterium Oscillatoria sp. Strain Miami BG7. AB - Heterocystous filamentous cyanobacterium Anabaena cylindrica B629 and nonheterocystous filamentous cyanobacterium Oscillatoria sp. strain Miami BG7 were cultured in media with N(2) as the sole nitrogen source; and activities of oxygen-dependent hydrogen uptake, photohydrogen production, photooxygen evolution, and respiration were compared amperometrically under the same or similar experimental conditions for both strains. Distinct differences in these activities were observed in both strains. The rates of hydrogen photoproduction and hydrogen accumulation were significantly higher in Oscillatoria sp. strain BG7 than in A. cylindrica B629 at every light intensity tested. The major reason for the difference was attributable to the fact that the heterocystous cyanobacterium had a high rate of oxygen-dependent hydrogen consumption activity and the nonheterocystous cyanobacterium did not. The activity of oxygen photoevolution and respiration also contributed to the difference. Oscillatoria sp. strain BG7 had lower O(2) evolution and higher respiration than did A. cylindrica B629. Thus, the effect of O(2) on hydrogen photoproduction was minimized in Oscillatoria sp. strain BG7. PMID- 16346851 TI - Methane fermentation of ferulate and benzoate: anaerobic degradation pathways. AB - The anaerobic biodegradation of ferulate and benzoate in stabilized methanogenic consortia was examined in detail. Up to 99% of the ferulate and 98% of the benzoate were converted to carbon dioxide and methane. Methanogenesis was inhibited with 2-bromoethanesulfonic acid, which reduced the gas production and enhanced the buildup of intermediates. Use of high-performance liquid chromatography and two gas chromatographic procedures yielded identification of the following compounds: caffeate, p-hydroxycinnamate, cinnamate, phenylpropionate, phenylacetate, benzoate, and toluene during ferulate degradation; and benzene, cyclohexane, methylcyclohexane, cyclohexanecarboxylate, cyclohexanone, 1-methylcyclohexanone, pimelate, adipate, succinate, lactate, heptanoate, caproate, isocaproate, valerate, butyrate, isobutyrate, propionate, and acetate during the degradation of either benzoate or ferulate. Based on the identification of the above compounds, more complete reductive pathways for ferulate and benzoate are proposed. PMID- 16346852 TI - PA-1, a Versatile Anaerobe Obtained in Pure Culture, Catabolizes Benzenoids and Other Compounds in Syntrophy with Hydrogenotrophs, and P-2 plus Wolinella sp. Degrades Benzenoids. AB - Methanogenic enrichments catabolizing 13 mM phenylacetate or 4 mM phenol were established at 37 degrees C, using a 10% inoculum from a municipal anaerobic digester. By using agar roll tubes of the basal medium plus 0.1% yeast extract-25 mM fumarate, a hydrogenotrophic lawn of Wolinella succinogenes and phenol or phenylacetate, strains P-2 and PA-1, respectively, were isolated in coculture with W. succinogenes. With the lawn deleted, PA-1 was isolated in pure culture. Strain P-2 is apparently a new species of anaerobic, motile, gram-negative, spindle-shaped, small rod that as yet has been grown only in coculture with W. succinogenes. It used phenol, hydrocinnamate, benzoate, and phenylacetate as energy sources. Product recovery by the coculture, per mole of phenol and 4.4 mol of fumarate used, included 2.03, 0.12, 0.08, and 3.23 mol, respectively, of acetate, propionate, butyrate, and succinate. Carbon recovery was 75% and H recovery was 80%, although CO(2) and a few other possible products were not determined. That P-2 is an obligate proton-reducing acetogen and possible pathways for its degradation of phenol are discussed. Strain PA-1 is apparently a new species of anaerobic, motile, relatively small, gram-negative rod. It utilized compounds such as phenylacetate, hydrocinnamate, benzoate, phenol, resorcinol, gallate, 4-aminophenol, 2-aminobenzoate, pyruvate, Casamino Acids, and aspartate as energy sources in coculture with W. succinogenes. Per mole of phenylacetate and 1.44 mol of fumarate used, 1.04, 0.53, and 0.78 mol of acetate, propionate, and succinate, respectively, were recovered from the coculture. Only about 50% of the carbon and H were recovered. In coculture with Methanospirillum hungatei, 0.96 mol of acetate and 0.25 mol of methane were recovered per mol of pyruvate used; 0.90 mol of acetate and 0.33 mol of methane, per mol of fumarate used; 0.93 mol of acetate and 0.54 mol of methane, per mol of aspartate used; and 1.71 mol of acetate and 0.57 mol of methane, per mol of glucose used. Carbon and H recoveries, assuming CO(2) and ammonia were produced in stoichiometric amounts, were 97 and 98% for pyruvate, 72.5 and 82% for fumarate, 96.5 and 98% for aspartate, and 61.8 and 76% for glucose. No explanation such as contamination could be found for the fact that the coculture PA-1 plus Wolinella sp. did not use glucose; after growth with M. hungatei on pyruvate, however, the latter coculture used glucose. The PA-1 pure culture produced 0.86 mol of propionate per mol of succinate used during growth. PA-1 produced a small amount of H(2). Strain PA-1 is the most versatile anaerobic bacterium yet known that catabolizes monobenzenoids in the absence of electron acceptors such as sulfate or nitrate. PMID- 16346853 TI - Microbial decomposition of cellulose in acidifying lakes of South-central ontario. AB - The rate of cellulose breakdown and density of bacterial populations were measured in the epilimnetic sediments and water columns of lakes in central Ontario that differ in pH, alkalinity, and nutrient status and are particularly sensitive to acidic inputs from atmospheric decomposition. There was no significant difference in decomposition rate in either oxic or anoxic sediment when mean epilimnetic pHs were in the range 5.5 to 6.9. The importance of these findings for the breakdown of autochthonous detritus in Canadian Shield lakes is discussed. Furthermore, the results of these experiments, in which dyed strips of cellophane (regenerated cellulose) were used as substrate, were compared with results of earlier decomposition studies carried out with coarse litter (leaves, twigs). Acridine orange direct counts of bacteria in the top 1 cm of sediment ranged from 5.5 x 10 to 1.0 x 10 per g and in planktonic water samples from 1.1 x 10 to 1.8 x 10 per ml. Bacterial densities were significantly higher in both the shallow sediment (P < 0.01) and the water column (P < 0.05) of dystrophic lakes than at these sites in oligotrophic lakes. PMID- 16346854 TI - Effects of chemical speciation on the mineralization of organic compounds by microorganisms. AB - The mineralization of 1.0 to 100 ng each of four complexing compounds-oxalate, citrate, nitrilotriacetate (NTA), and EDTA-per ml was tested in media prepared in accordance with equilibrium calculations by a computer program so that the H, Ca, Mg, Fe, or Al complex (chemical species) was predominant. Sewage microorganisms mineralized calcium citrate more rapidly than iron, aluminum, or hydrogen citrate, and magnesium citrate was degraded slowest. Aluminum, hydrogen, and iron oxalates were mineralized more rapidly than calcium oxalate, and magnesium oxalate was decomposed slowest. Sewage microorganisms mineralized calcium NTA but not aluminum, magnesium, hydrogen, or iron NTA or any of the EDTA complexes. Pseudomonas sp. mineralized calcium and iron citrates but had no activity on hydrogen, aluminum, or magnesium citrate. Pseudomonas pseudoalcaligenes mineralized calcium, iron, hydrogen, and aluminum citrates but had little activity on magnesium citrate. Pseudomonas alcaligenes used calcium, iron, hydrogen, and aluminum oxalates readily, but it used magnesium oxalate at a slower rate. Listeria sp. destroyed calcium NTA but had no effect on hydrogen, iron, or magnesium NTA. Increasing the Ca concentration in the medium enhanced the breakdown of NTA by Listeria sp. The different activities of the bacterial isolates were not a result of the toxicity of the complexes or the lack of availability of a nutrient element. NTA mineralization was not enhanced by the addition of Ca to Beebe Lake water, but it was enhanced when Ca and an NTA degrading inoculum were added to water from an oligotrophic lake. The data show that chemical speciation influences the mineralization of organic compounds by naturally occurring microbial communities and by individual bacterial populations. PMID- 16346855 TI - Enumeration, isolation, and characterization of n(2)-fixing bacteria from seawater. AB - Marine pelagic N(2)-fixing bacteria have not, in general, been identified or quantified, since low or negligible rates of N(2) fixation have been recorded for seawater when blue-green algae (cyanobacteria) are absent. In the study reported here, marine N(2)-fixing bacteria were found in all samples of seawater collected and were analyzed by using a most-probable-number (MPN) method. Two different media were used which allowed growth of microaerophiles, as well as that of aerobes and facultative anaerobes. MPN values obtained for N(2)-fixing bacteria ranged from 0.4 to 1 x 10 per liter for water collected off the coast of Puerto Rico and from 2 to 5.5 x 10 per liter for Chesapeake Bay water. Over 100 strains of N(2)-fixing bacteria were isolated from the MPN tubes and classified, yielding four major groups of NaCl-requiring bacteria based on biochemical characteristics. Results of differential filtration studies indicate that N(2) fixing bacteria may be associated with phytoplankton. In addition, when N(2) fixing bacteria were inoculated into unfiltered seawater and incubated in situ, nitrogenase activity could be detected within 1 h. However, no nitrogenase activity was detected in uninoculated seawater or when bacteria were incubated in 0.2-mum-filtered (phytoplankton-free) seawater. The ability of these isolates to fix N(2) at ambient conditions in seawater and the large variety of N(2)-fixing bacteria isolated and identified lead to the conclusion that N(2) fixation in the ocean may occur to a greater degree than previously believed. PMID- 16346856 TI - Formation of Filaments by Pseudomonas putida. AB - When Pseudomonas putida 40 was grown on a variety of liquid media in which oxygen became a limiting factor during growth, the latter stages of growth involved the elongation of cells without septation, which can result in the complete filamentation of the culture (up to several hundred micrometers long). The filaments appeared to consist of a chain of protoplasts within a common sacculus. Later these filaments were capable of a rapid fragmentation by septation to give a population of ordinary rods with a corresponding increase in the number of viable particles but no appreciable change in total bacterial mass. Filamentation did not occur if slow growth rates were maintained by restriction of oxygen availability from the beginning of growth. In complex media filaments were not formed during growth on 1% peptone alone, but the addition of 0.1 M phosphate or 6.6 x 10 M EDTA induced extensive filamentation that was reversed by the addition of 6.6 x 10 M Mg. In minimal media a much higher Mg concentration than that required for active growth or present in the complex media was usually required for filamentation. A very narrow range of Mg concentration promoted filamentation, and this optimum differed markedly depending on the carbon source used. Other medium variations which influenced the level of filamentation are reported. We found that most strains of P. putida (including the neotype strain) and P. fluorescens gave filaments under the conditions developed with strain 40, whereas several strains of P. aeruginosa failed to give filaments on the same media. PMID- 16346857 TI - Oxygen Responses and Mat Formation by Beggiatoa spp. AB - The behavioral response of single Beggiatoa sp. filaments moving on a gas permeable membrane was studied by the combined use of microscopy and oxygen microelectrodes during controlled oscillations of oxygen tension. The bacteria reacted to increasing oxygen by reversing the direction of movement. The same step-up phobic response to oxygen was observed when a filament tip or loop glided into a stable microgradient of increasing oxygen. The response was sensitive to a change in oxygen tension of <5% of air saturation min. The response time was 20 to 50 s. Frequently, only part of the filament responded, which led to the formation of sharp bends, loops, and coils. This partial response facilitated the positioning of the long filaments within the narrow O(2)-H(2)S interface. The structure of whole Beggiatoa mats on sediment surfaces varied from loose to dense in relation to shallow or steep oxygen gradients in the 0.3- to 2-mm-thick, unstirred boundary layer. In an illuminated sediment Beggiatoa spp. lived together with photosynthetic organisms and migrated vertically in accordance with light/dark variations. The combined effect of phobic responses to light and oxygen can explain this migration. PMID- 16346858 TI - Microbial Penetration through Nutrient-Saturated Berea Sandstone. AB - Penetration times and penetration rates for a motile Bacillus strain growing in nutrient-saturated Berea sandstone cores were determined. The rate of penetration was essentially independent of permeabilities above 100 mdarcys and rapidly declined for permeabilities below 100 mdarcys. It was found that these penetration rates could be grouped into two statistically distinct classes consisting of rates for permeabilities above 100 mdarcys and rates for those below 100 mdarcys. Instantaneous penetration rates were found to be zero order with respect to core length for cores with permeabilities above 100 mdarcys and first order with respect to core length for cores with permeabilities below 100 mdarcys. The maximum observed penetration rate was 0.47 cm . h, and the slowest was 0.06 cm . h; however, these rates may be underestimates of the true penetration rate, since the observed rates included the time required for growth in the flask as well as the core. The relationship of penetration time to the square of the length of the core suggested that cells penetrated high permeability cores as a band and low-permeability cores in a diffuse fashion. The motile Enterobacter aerogenes strain penetrated Berea sandstone cores three to eight times faster than did the nonmotile Klebsiella pneumoniae strain when cores of comparable length and permeability were used. A penetration mechanism based entirely on motility predicted penetration times that were in agreement with the observed penetration times for motile strains. The fact that nonmotile strains penetrated the cores suggested that filamentous or unrestricted growth, or both, may also be important. PMID- 16346859 TI - Relationship between Rapid, Firm Adhesion and Long-Term Colonization of Roots by Bacteria. AB - For rhizobacteria to exert physiological effects on plant growth, the bacteria must first effectively colonize the root surface. To examine the relationship between long-term colonization of root systems and adherence to roots in the short term, a binding assay was developed. Adherence was determined by incubating roots of intact radish seedlings with bacteria, washing and homogenizing the roots, and dilution plating the resulting homogenate. Irreversible binding of bacteria was rapid, reaching half-maximum by 5 min. All of the rhizosphere bacteria tested showed similar, concentration-dependent binding (ranging from 10 to 10 CFU/ml), as well as long-term colonization of radish roots under sterile conditions. Escherichia coli, which is not a root colonizer, showed about 10-fold less binding, but still demonstrated concentration-dependent binding and rapid kinetics of adherence at high concentrations (10 to 10 CFU/ml). The bacteria tested were very different with respect to source or habitat and plant response, yet they showed similar concentration-dependent binding. There was no correlation between the relative hydrophobicities of the cell surfaces of strains and the adherence of the strains to roots. Binding of Pseudomonas fluorescens E6-22 was promoted by divalent cations (Ca and Mg) at concentrations of 5 to 10 mM, whereas monovalent cations (Na and K) had little effect; electrostatic phenomena may partially explain adherence in the short term, an important prelude to long-term colonization of root surfaces. PMID- 16346860 TI - Inoculant Production with Diluted Liquid Cultures of Rhizobium spp. and Autoclaved Peat: Evaluation of Diluents, Rhizobium spp., Peats, Sterility Requirements, Storage, and Plant Effectiveness. AB - Fully grown broth cultures of various fast- and slow-growing rhizobia were deliberately diluted with various diluents before their aseptic incorporation into autoclaved peat in polypropylene bags (aseptic method) or mixed with the peat autoclaved in trays (tray method). In a factorial experiment with the aseptic method, autoclaved and irradiated peat samples from five countries were used to prepare inoculants with water-diluted cultures of three Rhizobium spp. When distilled water was used as the diluent, the multiplication and survival of rhizobia in the peat was similar to that with diluents having a high nutrient status when the aseptic method was used. In the factorial experiment, the mean viable counts per gram of inoculant were log 9.23 (strain TAL 102) > log 8.92 (strain TAL 82) > log 7.89 (strain TAL 182) after 24 weeks of storage at 28 degrees C. The peat from Argentina was the most superior for the three Rhizobium spp., with a mean viable count of log 9.0 per g at the end of the storage period. The quality of inoculants produced with diluted cultures was significantly (P = 0.05) better with irradiated than with autoclaved peat, as shown from the factorial experiment. With the tray method, rhizobia in cultures diluted 1,000 fold or less multiplied and stored satisfactorily in the presence of postinoculation contaminants, as determined by plate counts, membrane filter immunofluorescence, and plant infection procedures. All strains of rhizobia used in both the methods showed various degrees of population decline in the inoculants when stored at 28 degrees C. Fast- and slow-growing rhizobia in matured inoculants produced by the two methods showed significant (P < 0.01) decline in viability when stored at 4 degrees C, whereas the viability of some strains increased significantly (P < 0.01) at the same temperature. The plant effectiveness of inoculants produced with diluted cultures and autoclaved peat did not differ significantly from that of inoculants produced with undiluted cultures and gamma-irradiated peat. PMID- 16346861 TI - Evidence for Separate Adhesion Mechanisms for Hydrophilic and Hydrophobic Surfaces in Vibrio proteolytica. AB - The proteolytic enzymes pronase, trypsin, and chymotrypsin and the surfactant Triton X-100 inhibited attachment of Vibrio proteolytica to the hydrophobic substratum polystyrene by >97%. These treatments had no effect on attachment to hydrophilic substrata such as glass or tissue culture dishes. Both pronase and Triton X-100 effected the removal of previously attached cells from polystyrene but not from hydrophilic surfaces. Removal of cells from polystyrene by pronase left material (which we have termed footprints) that stained with the protein specific stain Hoechst 2495 but not with the DNA-specific stain Hoechst 33342. Pronase treatment also caused a significant decrease in cell surface hydrophobicity as determined by phase partitioning in hexane or petroleum ether. Collectively, these results imply the existence of separate mechanisms for the adhesion of V. proteolytica to hydrophilic and hydrophobic substrata and suggest a role for protein in the latter mechanism. PMID- 16346862 TI - Carbon mineralization in acidic, xeric forest soils: induction of new activities. AB - Carbon mineralization was examined in Lakehurst and Atsion sands collected from the New Jersey Pinelands and in Pahokee muck from the Everglades Agricultural Area. Objectives were (i) to estimate the carbon mineralization capacities of acidic, xeric Pinelands soils in the absence of exogenously supplied carbon substrate (nonamended carbon mineralization rate) and to compare these activities with those of agriculturally developed pahokee muck, and (ii) to measure the capacity for increased carbon mineralization in the soils after carbon amendment. In most cases, nonamended carbon mineralization rates were greater in samples of the acid- and moisture-stressed Pinelands soils than in Pahokee muck collected from a fallow (bare) field. Carbon amendment resulted in augmented catabolic activity in Pahokee muck samples, suggesting that the microbial community was carbon limited in this soil. With many of the substrates, no stimulation of the catabolic rate was detected after amendment of Pinelands soils. This was documented by the observation that amendment of Pahokee muck with an amino acid mixture, glucose, or acetate resulted in a 3.0-, 3.9-, or 10.5-fold stimulation of catabolic activity, respectively, for the added substrate. In contrast, amendment of the Pinelands soils resulted in increased amino acid and acetate catabolic rates in Lakehurst sand and increased acetate metabolism only in Atsion sand. Other activities were unchanged. The increased glucose respiration rates resulted from stimulation of existing microbial activity rather than from microbial proliferation since no change in the microbial growth rate, as estimated by the rate of incorporation of C-labeled acetate into cell membranes, occurred after glucose amendment of the soils. A stimulation of microbial growth rate was recorded with glucose-amended Lakehurst sand collected from the B horizon. PMID- 16346863 TI - Growth kinetics of attached iron-oxidizing bacteria. AB - A model of growth and substrate utilization for ferrous-iron-oxidizing bacteria attached to the disks of a rotating biological contactor was developed and tested. The model describes attached bacterial growth as a saturation function in which the rate of substrate utilization is determined by a maximum substrate oxidation rate constant (P), a half-saturation constant (K(s)), and the concentration of substrate within the rotating biological contactor (S(1)). The maximum oxidation rate constant was proportional to flow rate, and the substrate concentration in the reactor varied with influent substrate concentration (S(0)). The model allowed the prediction of metabolic constants and included terms for both constant and growth-rate-dependent maintenance energies. Estimates for metabolic constants of the attached population of acidophilic, chemolithotrophic, iron-oxidizing bacteria limited by ferrous iron were: maximum specific growth rate (mu(max)), 1.14 h; half-saturation constant (K(s)) for ferrous iron, 54.9 mg/liter; constant maintenance energy coefficient (m(1)), 0.154 h; growth-rate dependent maintenance energy coefficient (m'), 0.07 h; maximum yield (Y(g)), 0.063 mg of organic nitrogen per mg of Fe(II) oxidized. PMID- 16346864 TI - Immobilized Clostridium acetobutylicum P262 Mutants for Solvent Production. AB - The production of acetone, butanol, and ethanol by two immobilized, sporulation deficient (spo) Clostridium acetobutylicum P262 mutants which were held in the solventogenic phase was investigated. The spoA2 mutant, which was an early sporulation mutant and did not form a forespore septum, produced higher solvent yields than did the spoB mutant which was a late-sporulation mutant and was blocked at a stage after forespore septum formation. The spoA2 mutant was also granulose and capsule negative. In a conventional batch fermentation, the wild type strain produced 15.44 g of solvents per liter after 50 h at a productivity of 7.41 g of solvents per liter per day. The spoA2 mutant produced 15.42 g of solvents per liter at a productivity of 72.4 g of solvents per liter per day, with a retention time of 2.4 h in a continuous immobilized cell system employing a fluidized bed reactor. This represents a major advance, since the immobilization of wild-type cells showed similar increases in productivity but a ca. fivefold reduction in final product concentrations. PMID- 16346865 TI - Microbial manganese reduction by enrichment cultures from coastal marine sediments. AB - Manganese reduction was catalyzed by enrichment cultures of anaerobic bacteria obtained from coastal marine sediments. In the absence of oxygen, these enrichment cultures reduced manganates when grown on either lactate, succinate, or acetate in both sulfate-free and sulfate-containing artificial seawaters. Sodium azide as well as oxygen completely inhibited microbial manganese reduction by these enrichment cultures, whereas molybdate had no effect on them. The addition of nitrate to the medium slightly decreased the rate of Mn production by these enrichment cultures. These findings are consistent with the hypothesis that the manganese-reducing organisms in these enrichment cultures use manganates as terminal electron acceptors and couple manganese reduction in some way to the oxidation of organic matter. PMID- 16346866 TI - Sulfate-reducing bacteria: principal methylators of mercury in anoxic estuarine sediment. AB - Substrate-electron acceptor combinations and specific metabolic inhibitors were applied to anoxic saltmarsh sediment spiked with mercuric ions (Hg) in an effort to identify, by a direct approach, the microorganisms responsible for the synthesis of hazardous monomethylmercury. 2-Bromoethane sulfonate (30 mM), a specific inhibitor of methanogens, increased monomethylmercury synthesis, whereas sodium molybdate (20 mM), a specific inhibitor of sulfate reducers, decreased Hg methylation by more than 95%. Anaerobic enrichment and isolation procedures yielded a Desulfovibrio desulfuricans culture that vigorously methylated Hg in culture solution and also in samples of presterilized sediment. The Hg methylation activity of sulfate reducers is fully expressed only when sulfate is limiting and fermentable organic substrates are available. To date, sulfate reducers have not been suspected of Hg methylation. Identification of these bacteria as the principal methylators of Hg in anoxic sediments raises questions about the environmental relevance of previous pure culture-based methylation work. PMID- 16346868 TI - Plasmid DNA in Strains of Pediococcus cerevisiae and Pediococcus pentosaceus. AB - Five parental strains of Pediococcus were examined for plasmid content. Each strain contained three to six resident plasmids, ranging in size from 4.5 to 39.5 megadaltons. A bacteriocin-like substance produced by Pediococcus cerevisiae FBB63 was tentatively linked to a 10.5-megadalton plasmid after being cured with novobiocin. PMID- 16346867 TI - Bacterial biomass, metabolic state, and activity in stream sediments: relation to environmental variables and multiple assay comparisons. AB - Bacterial biomass, metabolic condition, and activity were measured over a 16 month period in the surface sediments of the following four field sites with differing dissolved organic matter regimes: a woodlot spring seep, a meadow spring seep, a second-order stream, and a third-order stream. Total bacterial biomass was measured by lipid phosphate and epifluorescence microscopic counts (EMC), and viable biomass was measured by C most probable number, EMC with 2-(p iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride reduction, and ATP. Bacterial metabolic condition was determined from the percentage of respiring cells, poly-beta-hydroxybutyrate concentrations, and adenylate energy charge. Activity measures included C-lipid synthesis, P-phospholipid synthesis, the rate of uptake of algal lysate dissolved organic carbon, and respiration, from which biosynthesis was calculated (dissolved organic carbon uptake corrected for respiration). Total bacterial biomass (from EMC) ranged from 0.012 to 0.354 mug of C/mg of dry sediment and was usually lowest in the third-order stream. The percentage of cells respiring was less than 25% at all sites, indicating that most bacteria were dormant or dead. Adenylate energy charge was measured only in the third-order stream and was uniformly low. Poly-beta-hydroxybutyrate concentrations were greater in the woodlot spring seep than in the second- and third-order streams. Uptake of algal lysate dissolved organic carbon ranged from undetectable levels to 166 mg of C . m . h. Little community respiration could be attributed to algal lysate metabolism. Phospholipid synthesis ranged from 0.006 to 0.354 pmol . mg of dry sediment . h. Phospholipid synthesis rates were used to estimate bacterial turnover at the study sites. An estimated 375 bacterial generations per year were produced in the woodlot spring seep, and 67 per year were produced in the third-order stream. PMID- 16346869 TI - Determination of the hydrogenase status of individual legume nodules by a methylene blue reduction assay. AB - We adapted a method for the rapid screening of colonies of free-living Rhizobium japonicum for hydrogenase activity to determine the hydrogenase status of individual soybean nodules. Crude bacteroid suspensions from nodules containing strains known to be hydrogen uptake positive (Hup) caused a localized decolorization of filter paper disks, whereas suspensions from nodules arising from inoculation with hydrogen uptake-negative (Hup) mutants or strains did not decolorize the disks. The reliability of the method was demonstrated by its successful application to 29 slow-growing rhizobia. The Hup phenotype on methylene blue filters agreed with that determined amperometrically with either methylene blue or oxygen as the electron acceptor. PMID- 16346870 TI - Occurrence of beta-Aminoglutaric Acid in Marine Bacteria. AB - beta-Aminoglutaric acid, a nonprotein amino acid isomer of glutamic acid, was found in the free amino acid pool of a marine bacterium, Alteromonas luteoviolacea. It was also found in a mixed culture of fermenting bacteria enriched from an anoxic marine sediment. PMID- 16346871 TI - Low-Temperature Production of Urocanic Acid by Spoilage Bacteria Isolated from Mahimahi (Coryphaena hippurus). AB - Urocanic acid production was studied in 166 bacterial cultures isolated from mahimahi which had been stored for 14 days on ice in seawater. After 4 days of incubation at 10 degrees C in histidine-supplemented Trypticase soy broth (BBL Microbiology Systems, Cockeysville, Md.), urocanic acid was measured by thin layer chromatography. Fifty-eight of the cultures were positive for urocanic acid, with the eight most active isolates producing ca. 10 mg/100 ml. These observations, coupled with a report that urocanic acid may be the predominant histidine metabolite in fish held at low temperatures, all suggest that urocanic acid may be a useful alternative to histamine as a spoilage index in scombroid and other fish that are rich in endogenous histidine. PMID- 16346872 TI - Comparison of Media for the Isolation of Cryptococcus neoformans. AB - Four media, Staib's Guizotia abyssinica, trypan blue, and Staib's with 2 and 10 mg of methyl violet per liter, were compared for the selective and differential isolation of Cryptococcus neoformans from environmental samples. Trypan blue medium allowed for the differentiation of C. neoformans colonies from Candida albicans colonies several days earlier than did Staib's medium. The addition of methyl violet to Staib's medium was found to be inhibitory to some strains of all species tested. Diphenyl in Staib's medium inhibited the growth of 30 strains of C. neoformans and C. albicans. PMID- 16346873 TI - Toxigenic potential of some fusarium isolates from southeast Asia. AB - The isolation and characterization of 10 isolates of six Fusarium spp. from plant and soil samples collected in Southeast Asia is reported. The ability of these isolates to produce trichothecenes both in liquid cultures (CZ, GYEP, and MYRO) and on rice was assessed, and their toxigenic potential was examined by skin assay and gavage studies with culture filtrates. Although culture filtrates of all the isolates caused minor damage to test animals, only that of F. equiseti DAOM 189762 produced trichothecenes. PMID- 16346874 TI - Multiphasic kinetics for transformation of methyl parathion by flavobacterium species. AB - Transformation rates of an insecticide, methyl parathion, in pure cultures of Flavobacterium sp. followed multiphasic kinetics involving at least two systems (I and II). System I was a high-affinity, low-capacity system, and system II was a low-affinity, high-capacity system. Data from rate experiments suggested that metabolites formed via system II inhibited system I such that only one system operated at a time. System I operated at approximately 20 mug liter and less; system II operated at approximately 4 mg liter and less. These results show that xenobiotic chemicals, like naturally occurring substrates, can be transformed via multiple uptake and transformation systems even by a pure culture. Furthermore, computer simulation models of pollutant transformation rates based on kinetic constants determined in this study show that large errors can occur in predicted rates when the multiphasicity of kinetics is neglected. PMID- 16346875 TI - Selective delignification of aspen wood blocks in vitro by three white rot basidiomycetes. AB - Aspen wood blocks were selectively delignified in the laboratory by Ischnoderma resinosum, Poria medulla-panis, and Xylobolus frustulatus. After 8 weeks only the outer surfaces of wood blocks were selectively delignified. The percentages of weight loss obtained after 4, 8, and 12 weeks showed that decay occurred at a relatively constant rate. Selectively delignified wood could be identified by using scanning electron microscopy only when lignin had been extensively removed from cell walls. X. frustulatus was able to form pockets of delignified wood throughout blocks after 12 weeks. PMID- 16346876 TI - Glucose catabolism in strains of acidophilic, heterotrophic bacteria. AB - Pathways of glucose catabolism, potentially operational in six strains of obligately aerobic, acidophilic bacteria, including Acidiphilium cryptum strain Lhet2, were investigated by short-term radiorespirometry and enzyme assays. Short term radiorespirometry was conducted at pH 3.0 with specifically labeled [C]glucose. The high rate and yield of C-1 oxidized to CO(2) indicated that the Entner-Doudoroff, pentose phosphate, or both pathways were operational in all strains. Apparent nonequivalent yields of CO(2) from C-1 and estimated CO(2) from C-4 (C-1 > C-4) were suggestive of simultaneous glucose catabolism by both pathways in all strains tested. Variation in the relative contribution of the two pathways of glucose catabolism appears to account for observed strain differences. Calculation of the actual percent pathway participation was not feasible. Enzyme assays were completed with crude extracts of glucose-grown cells to substantiate the results obtained by radiorespirometry. The key enzymes of the pentose phosphate pathway (6-phosphogluconate dehydrogenase) and the Entner Doudoroff pathway (2-keto-3-deoxy-6-phosphogluconate aldolase and 6 phosphogluconate dehydrase) were present in all strains examined (PW2, Lhet2, KLB, OP, and QBP). However, none of the strains exhibited detectable levels of the key enzyme of the Embden-Meyerhof-Parnas pathway, 6-phosphofructokinase. All strains contained glucose-6-phosphate dehydrogenase and fructose bisphosphate aldolase. The results of the enzyme study supported the contention that the pentose phosphate and Entner-Doudoroff pathways are operational for glucose catabolism in the acidophilic heterotrophs, and that the Embden-Meyerhof-Parnas pathway is apparently absent. PMID- 16346877 TI - Characterization of subsurface bacteria associated with two shallow aquifers in oklahoma. AB - The bacterial microflora of two shallow aquifers (saturated subsurface zones) in Oklahoma was characterized by direct observation with light and electron microscopy, by plating, and by examination of colony morphology and distribution. Isolated bacterial strains were also examined. Total cell counts varied only slightly (2.9 x 10 to 9.8 x 10 g [dry wt]) from sample to sample, whereas colony counts varied widely (6.3 x 10 to 6.5 x 10 CFU g [dry wt]). Colony counts on nutritionally rich media were lower than on low-nutrient media, especially in samples from the saturated zone. The variety of colony types growing on nutritionally rich media decreased with increasing depth and saturation. Colony counts of anaerobic bacteria also decreased with depth but were at least 100-fold lower than aerobic counts on most media. Cell morphologies of bacteria grown aerobically on plates included short rods, cocci, and actinomycete-like forms. Direct light microscopic observation of sediments revealed short, rod-shaped, and coccoid bacterial cells; endospores, actinomycete spores, and eucaryotic forms were not observed by light microscopy. Electron microscopic observation of bacteria released from the samples revealed that 85 to 90% of them were coccoid, gram-positive, Arthrobacter-like organisms, some of which were dividing or contained completed division septa; other types of gram-positive and gram negative bacteria were present in lower numbers. Isolated bacterial strains were able to grow on both nutritionally rich and low-nutrient media. A higher proportion of gram-negative organisms was isolated than gram-positive organisms. Most of the isolates were capable of storing polyphosphate, poly-beta hydroxybutyrate, or polysaccharide. The results of this study suggest that the microbial population of these two shallow aquifers is dominated by aerobic, nutritionally versatile bacteria that can subsist on low concentrations of organic compounds without forming specialized resting cells. Other types of microorganisms, such as facultatively anaerobic bacteria and microeucaryotes, may also be present, but they represent only a small fraction of the microflora. PMID- 16346878 TI - Sulfate-Dependent Interspecies H(2) Transfer between Methanosarcina barkeri and Desulfovibrio vulgaris during Coculture Metabolism of Acetate or Methanol. AB - We compared the metabolism of methanol and acetate when Methanosarcina barkeri was grown in the presence and absence of Desulfovibrio vulgaris. The sulfate reducer was not able to utilize methanol or acetate as the electron donor for energy metabolism in pure culture, but was able to grow in coculture. Pure cultures of M. barkeri produced up to 10 mumol of H(2) per liter in the culture headspace during growth on acetate or methanol. In coculture with D. vulgaris, the gaseous H(2) concentration was 2 degrees C. PMID- 16346886 TI - Characterization of Leuconostoc oenos Isolated from Oregon Wines. AB - This study was designed to characterize isolates of Leuconostoc species from Oregon wines. Gram-positive cocci were isolated, and their biochemical properties and abilities to decompose malic acid were determined. All of the isolates were heterofermentative, catalase negative, and facultatively anaerobic and occurred in pairs and chains. They produced acid from glucose, fructose, mannose, ribose, cellobiose, trehalose, and salicin but not from sucrose or lactose. They did not produce ammonia from arginine or dextran from sucrose. They grew at pH values of less than 4 and in 10% ethanol. Most but not all strains produced lactic acid and carbon dioxide from malic acid, as determined by paper chromatography and respirometry, respectively. These malolactic bacteria were considered to be strains of Leuconostoc oenos. We compared these isolates with reference strains for relative growth at pH values of 4.0, 3.5, 3.0, and 2.8 at 22 degrees C. The isolates were similar in their growth responses at the two highest pH levels. At pH 3.0 and 2.8, however, the strains failed to grow but revealed variable abilities to dissimilate malic acid. PMID- 16346887 TI - Ultrastructure of Sporulating Bacillus larvae in a Broth Medium. AB - An electron microscopic study of sporulation of Bacillus larvae, a honeybee pathogen, in TMYGP broth (D. W. Dingman and D. P. Stahly, Appl. Environ. Microbiol. 46:860-869, 1983) was conducted. No parasporal structures were evident in the sporangial cytoplasm. The stages of sporulation were similar to those observed in other sporeformers. A rather unusual inner coat layer consisting of seven lamellae was apparent. PMID- 16346888 TI - Genetic Recombination and Transformation in Protoplasts of Thermomonospora fusca. AB - Protoplasts were produced from the thermophilic actinomycete Thermomonospora fusca and were regenerated to 0.1% of the direct count on regeneration agar. Recombination after protoplast fusion was demonstrated with drug-resistant mutants of T. fusca YX. A single thiostrepton-resistant colony was isolated after transformation of T. fusca YX with the streptomycete vector pIJ702, providing the first evidence for transformation in the genus Thermomonospora and suggesting that some mesophilic streptomycete genes can be expressed in thermophilic actinomycetes. Of 20 thermophilic actinomycete strains isolated from self-heated composts, 3 were found to harbor native plasmid DNA, providing potential sequences for the development of Thermomonospora-Streptomyces shuttle vectors. PMID- 16346889 TI - Anaerobic Production of a Biosurfactant by Bacillus licheniformis JF-2. AB - Bacillus licheniformis JF-2 anaerobically produced a biosurfactant when grown in a glucose-mineral salts medium containing yeast extract and NaNO(3). Surface tension of the medium was reduced from 70 to 74 mN/m to as low as 28 mN/m due to the production of an anionic biosurfactant. PMID- 16346890 TI - Response of Sesbania grandiflora to Inoculation of Soil with Vesicular-Arbuscular Mycorrhizal Fungi. AB - A greenhouse experiment was conducted to determine the influence of two tropical isolates of Glomus fasciculatum and Glomus mosseae on the nutrient uptake and growth of Sesbania grandiflora. Inoculation of sterile soil with the fungi significantly improved growth and nutrient uptake by S. grandiflora, but the response of the legume was markedly better when the soil was inoculated with G. fasciculatum than when it was inoculated with G. mosseae. Nutrient uptake and growth of S. grandiflora in nonsterile soil was also significantly stimulated by inoculation, but the legume did not respond differently to the two endophytes under this condition. PMID- 16346891 TI - Measurement of Microbial Nucleic Acid Synthesis and Specific Growth Rate by PO(4) and [H]Adenine: Field Comparison. AB - We compared two radiotracer methods for measuring total microbial nucleic acid synthesis and specific growth rate. Using a sample from an oligotrophic environment, we found that there was excellent agreement between the two methods. PMID- 16346892 TI - Rhizobium sp. Degradation of Legume Root Hair Cell Wall at the Site of Infection Thread Origin. AB - Using a new microinoculation technique, we demonstrated that penetration of Rhizobium sp. into the host root hair cell occurs at 20 to 22 h after inoculation. It did this by dissolving the cell wall maxtrix, leaving a layer of depolymerized wall microfibrils. Colony growth pressure "stretched" the weakened wall, forming a bulge into an interfacial zone between the wall and plasmalemma. At the same time vesicular bodies, similar to plasmalemmasomes, accumulated at the penetration site in a manner which parallels host-pathogen systems. PMID- 16346893 TI - Production and Regeneration of Thiobacillus ferrooxidans Spheroplasts. AB - We investigated the production and regeneration of Thiobacillus ferrooxidans spheroplasts. Regeneration frequencies of up to 90% reversion were obtained. Attempts to transform T. ferrooxidans spheroplasts with a recombinant T. ferrooxidans plasmid were unsuccessful. PMID- 16346894 TI - Growth of Natural Yeast Flora during the Fermentation of Inoculated Wines. AB - The growth of yeasts that occur naturally in grape juice was quantitatively examined during the fermentation of four wines that had been inoculated with Saccharomyces cerevisiae. Although S. cerevisiae dominated the wine fermentations, there was significant growth of the natural species Kloeckera apiculata, Candida stellata, Candida colliculosa, Candida pulcherrima, and Hansenula anomala. PMID- 16346896 TI - Gibberella fujikuroi Mutants Obtained with UV Radiation and N-Methyl-N'-Nitro-N Nitrosoguanidine. AB - [This corrects the article on p. 187 in vol. 49.]. PMID- 16346895 TI - Stem and Root Nodulation in Aeschynomene spp. AB - Nodulation ability of 15 Rhizobium strains isolated from root and stem nodules of tropical Aeschynomene species was studied on 20 different Aeschynomene species and four other legumes-Arachis hypogaea, Stylosanthes guianensis, Macroptilium atropurpureum, and Sesbania rostrata. The results of this investigation showed that Aeschynomene species could be divided into three groups according to the cross-inoculation group concept. PMID- 16346897 TI - Observations of barophilic microbial activity in samples of sediment and intercepted particulates from the demerara abyssal plain. AB - To better understand the ecological significance of pressure effects on bacteria in the abyssobenthic boundary layer, experimental suspensions of sediments and sinking particulates were prepared from samples collected in boxcore and bottom moored sediment traps at two stations (depth, 4,470 and 4,850m) in the Demerara abyssal plain off the coast of Brazil. Replicate samples were incubated shipboard at 3 degrees C and at both atmospheric and deep-sea pressures (440 or 480 atm [4.46 x 10 or 4.86 x 10 kPa]) following the addition of [C]glutamic acid (<10 mug liter) or yeast extract (0.025%) and the antibiotic nalidixic acid (0.002%). In seven of the eight samples supplemented with isotope, a barophilic microbial response was detected, i.e., substrate incorporation and respiration were greater under in situ pressure than at 1 atm (101.3 kPa). In the remaining sample, prepared from a sediment trap warmed to 24 degrees C before recovery, pressure was observed to inhibit substrate utilization. Total bacterial counts by epifluorescence microscopy decreased with depth in each sediment core, as did utilization of glutamic acid. Significant percentages of the total bacterial populations in cold sediment trap samples (but not the prewarmed one or any boxcore sample) were abnormally enlarged and orange fluorescing after incubation with yeast extract and nalidixic acid under deep-sea conditions. Results indicated that in the deep sea, barophilic bacteria play a predominant role in the turnover of naturally low levels of glutamic acid, and the potential for intense microbial activity upon nutrient enrichment is more likely to occur in association with recently settled particulates, especially fecal pellets, than in buried sediments. PMID- 16346898 TI - Cellulose Decomposition and Associated Nitrogen Fixation by Mixed Cultures of Cellulomonas gelida and Azospirillum Species or Bacillus macerans. AB - Mixed cultures of Cellulomonas gelida plus Azospirillum lipoferum or Azospirillum brasilense and C. gelida plus Bacillus macerans were shown to degrade cellulose and straw and to utilize the energy-yielding products to fix atmospheric nitrogen. This cooperative process was followed over 30 days in sand-based cultures in which the breakdown of 20% of the cellulose and 28 to 30% of the straw resulted in the fixation of 12 to 14.6 mg of N per g of cellulose and 17 to 19 mg of N per g of g straw consumed. Cellulomonas species have certain advantages over aerobic cellulose-degrading fungi in being able to degrade cellulose at oxygen concentrations as low as 1% O(2) (vol/vol) which would allow a close association between cellulose-degrading and microaerobic diazotrophic microorganisms. Cultures inoculated with initially different proportions of A. brasilense and C. gelida all reached a stable ratio of approximately 1 Azospirillum/3 Cellulomonas cells. PMID- 16346899 TI - Starch-Ampicillin Agar for the Quantitative Detection of Aeromonas hydrophila. AB - Interest in Aeromonas hydrophila as a food-borne and human pathogen is increasing. Isolation media from the clinical laboratory were evaluated for food use and either did not give quantitative recovery of A. hydrophila or did not permit ready differentiation of A. hydrophila from the background microflora. A new medium was developed which permitted quantitative recovery of A. hydrophila from foods. The medium consisted of phenol red agar base (Difco Laboratories), soluble starch (10 g/liter), and ampicillin (10 mg/liter). All foods surveyed contained A. hydrophila. Foods sampled included red meats, chicken, raw milk, and seafood (fish, shrimp, scallops, crab, and oysters). The count of A. hydrophila at the time of purchase ranged from 1 x 10/g (lower limit of detection) to 5 x 10/g. In most instances, the count of A. hydrophila increased during 1 week of storage at 5 degrees C. The starch-ampicillin agar developed permitted rapid quantitative recovery of A. hydrophila from foods in the presence of very large numbers of competing microflora. PMID- 16346900 TI - Significance of Extracellular Protease for Growth of a Heterotrophic Bacterium, Aeromonas salmonicida. AB - The role of protease produced by a heterotrophic bacterium during growth was investigated with Aeromonas salmonicida, the pathogen of fish furunculosis, strain A-7301 and its protease-deficient mutant NTG-1 induced by mutagenesis. Strain A-7301 produced extracellular protease in a mixed amino acid medium (composed of Gly, Ala, Val, Ile, Leu, Thr, Ser, Cys, Met, Phe, Tyr, Lys, Arg, Pro, His, Try, Asp, Asn, Glu, and Gln at equal concentrations of 0.1 g/liter). Its multiplication rate was limited by the amounts of amino acids present, whereas strain NTG-1 showed no protease production despite considerable growth similar to that of A-7301. There was no difference between A-7301 and NTG-1 in amino acid requirements for growth, and seven amino acids (Gly, Ala, Val, Thr, Cys, Met, and His) were found to be indispensable. A defined level of the mixed amino acids (0.4 to 0.5 g/liter) was needed for A-7301 to initiate a large production of protease. Neither of the strains grew well in a casein medium, to which no amino acids were added. However, when a protease fraction obtained from extracellular products of A-7301 by DEAE-cellulose column chromatography was added, NTG-1 successfully reproduced in the casein medium. These results indicate that the extracellular protease plays an important role in supplying A. salmonicida cells with available amino acids as nutrients and that higher growth is closely associated with protease production which stimulates further reproduction. PMID- 16346901 TI - Pyrazines responsible for the potatolike odor produced by some serratia and cedecea strains. AB - The pyrazines responsible for the potatolike odor produced by some Serratia and Cedecea cultures were identified by gas-liquid chromatography (with flame ionization and thermoionic ionization detectors) and mass spectrometry. Alkylpyrazines were produced by the six strains studied irrespective of their odors. The major alkyl-methoxypyrazine produced by Cedecea davisae was 3-sec butyl-2-methoxypyrazine, and that produced by odoriferous Serratia strains (S. rubidaea, S. odorifera, and S. ficaria) was 3-isopropyl-2-methoxy-5-methyl pyrazine. Other pyrazines produced by some strains were 3-isopropyl-2 methoxypyrazine, 3-isobutyl-2-methoxypyrazine, 3-sec-butyl-2-methoxy-5 methylpyrazine, and 3-isobutyl-2-methoxy-6-methylpyrazine. Some of these pyrazines had not previously been found as natural products or to be produced by bacteria. PMID- 16346902 TI - Kinetics of biphenyl and polychlorinated biphenyl metabolism in soil. AB - The metabolism of C-labeled PCBs (polychlorinated biphenyls), which comprised the Aroclor 1242 mixture, was greatly enhanced by the addition of biphenyl (BP) to soil. After 49 days, only 25 to 35% of the original PCBs remained in the soil, and 48 to 49% was converted to CO(2) (including soil carbonates) in treatments enriched with BP; by contrast, 92% of the PCBs remained and less than 2% was converted to CO(2) in the unenriched control. Although the mineralization of PCBs in soils inoculated with Acinetobacter strain P6 was not greater than that in uninoculated BP-enriched soils, the initial and maximum mineralization rates and the disappearance of more highly chlorinated PCBs were greater with Acinetobacter strain P6. The mineralization of BP was consistent with kinetic models based upon linear-no growth and exponential growth; lower cell densities (<10/g) of BP oxidizing bacteria gave a better fit for exponential growth, whereas the highest cell density (10/g) gave a better fit for linear-no growth. The numbers of BP oxidizing bacteria declined exponentially upon depletion of the substrate. Since the mineralization of the chlorinated cometabolites was brought about by microorganisms (commensals) other than BP oxidizers, CO(2) production could not be fit to either of the two growth models. However, CO(2) production from the highest-density inoculum could be fit to a first-order (no-growth) sequential reaction series. Although the population dynamics of the commensals could not be determined, the rate-limiting step in the cometabolic-commensal metabolism of PCBs to CO(2) had to be the initial oxidation, since the rate of CO(2) production was directly related to the population density of BP oxidizers. PMID- 16346903 TI - Improvement of a Wine Saccharomyces cerevisiae Strain by a Breeding Program. AB - Hybridization by spore conjugation was used to develop new and improved wine yeasts of Saccharomyces cerevisiae. The procedure was achieved with diploid, homothallic strains with high sporulation frequency and high spore viability. The method was verified by crossing flocculent and non-H(2)S-forming strains. Single spore descendants of the hybrids were studied by tetrad analysis with regard to the aforementioned characters and the other two winemaking traits, i.e., ethanol production and fermentation rate. A highly flocculent, non-H(2)S-forming wine yeast strain with a high fermentation rate and high ethanol production was obtained. PMID- 16346904 TI - Xylanolytic Activity of Clostridium acetobutylicum. AB - Of 20 strains of Clostridium spp. screened, 17 hydrolyzed larch wood xylan. Two strains of Clostridium acetobutylicum, NRRL B527 and ATCC 824, hydrolyzed xylan but failed to grow on solid media with larch xylan as the sole carbon source; however, strain ATCC 824 was subsequently found to grow on xylan under specified conditions in a chemostat. These two strains possessed cellulolytic activity and were therefore selected for further studies. In cellobiose-limited continuous cultures, strain NRRL B527 produced maximum xylanase activity at pH 5.2. Strain ATCC 824 produced higher xylanase, xylopyranosidase, and arabinofuranosidase activities in chemostat culture with xylose than with any other soluble carbon source as the limiting nutrient. The activities of these enzymes were markedly reduced when the cells were grown in the presence of excess glucose. The xylanase showed maximum activity at pH 5.8 to 6.0 and 65 degrees C. The enzyme was stable on the alkaline side of pH 5.2 but was unstable below this pH value. The extracellular xylanolytic activity from strain ATCC 824 hydrolyzed 12% of the larch wood xylan during a 24-h incubation period, yielding xylose, xylobiose, and xylotriose as the major hydrolysis products. Strain ATCC 824, after being induced to grow in batch culture in xylan medium supplemented with a low concentration of xylose, failed to grow reproducibly in unsupplemented xylan medium. A mutant obtained by mutagenesis with ethyl methanesulfonate was able to grow reproducibly in batch culture on xylan. Both the parent strain and the mutant were able to grow with xylan as the sole source of carbohydrate in continuous culture with the pH maintained at either 5.2 or 6.0. Under these conditions, the cells utilized approximately 50% of the xylan. PMID- 16346905 TI - Biodegradation of N-nitrosodimethylamine in aqueous and soil systems. AB - N-Nitrosodimethylamine (NDMA) was mineralized by microorganisms in aqueous and soil systems. Initial rates of mineralization (micrograms per milliliter per day) were calculated for a wide range of initial concentrations of NDMA (micrograms per milliliter to picograms per milliliter). Log-log plots of the data were fitted with both linear and nonlinear least-squares analyses; however, linear models provided better fits for the kinetic data in all cases. The slopes of the linear fits were not significantly different than 1.0 (P < 0.05); thus, first order reaction kinetics were in effect over the range of concentrations tested, and saturation kinetics were not achieved. Rate constants (day) and total percent mineralized increased with decreasing initial concentrations of NDMA. Rates of mineralization were reduced in aqueous systems when supplemental carbon was available, whereas in soils, percentages of organic matter and supplemental carbon had little effect on rates of mineralization. Implications of these results for predictions of rates and threshold limits of mineralization activity in natural systems are discussed. A laboratory scale simulated trickling filter containing an activated charcoal bed provided a suitable environment for mineralization of NDMA at concentrations of 50 and 100 mug/ml on a continuous basis. NDMA was not toxic to natural populations of microorganisms at concentrations up to 10 mg/ml. Using high-pressure liquid chromatography coupled with radioactivity detection, we identified formaldehyde and methylamine as intermediates produced during the biodegradation of NDMA. PMID- 16346906 TI - GELRITE as an Agar Substitute for the Cultivation of Mesophilic Methanobacterium and Methanobrevibacter Species. AB - GELRITE was evaluated as a gelling agent for the growth of mesophilic Methanobacterium and Methanobrevibacter species. GELRITE was shown to be superior to agar in its gel strength and clarity. Viable cell counts and colony sizes of Methanobacterium species were greater on GELRITE-based medium compared with agar based medium. PMID- 16346907 TI - Passive diffusion technique for concentration of short-chain volatile Fatty acids from seawater. AB - A diffusion chamber is described which concentrates short-chain, volatile fatty acids from seawater while simultaneously separating them from interfering salts. The procedure relies on the passive diffusion of volatile compounds from acidified seawater samples and their subsequent absorption onto a base impregnated filter. The method is simple, efficient, and adaptable to most commonly used methods of volatile acid analysis. PMID- 16346908 TI - Regulation of Nodulation by Rhizobium meliloti 102F15 on Its Mutant Which Forms an Unusually High Number of Nodules on Alfalfa. AB - A mutant (WL3A150) of Rhizobium meliloti 102F51 that elicits an unusually high number of nodules on its host, alfalfa (Medicago sativa), supports the idea that the host may rely on early bacteroid development in the nodule or on metabolites produced in the infection thread as one of the signals to control further nodulation. This mutant was initially isolated because of its Fix phenotype. It consistently formed many more nodules than all the other Fix mutants isolated from strain 102F51 (a total of 11 mutants). Nodules formed by this mutant were small and white and were indistinguishable in appearance from nodules formed by the other Fix mutants. An ultrastructural study of the nodules, however, showed that this mutant, although forming numerous infection threads, failed to develop into bacteroids. The ability of the mutant to form an unusually high number of nodules coulde be suppressed in a time-dependent manner by the presence of the wild type. PMID- 16346909 TI - Maintenance of Different Mannitol Uptake Systems during Starvation in Oxidative and Fermentative Marine Bacteria. AB - The mannitol uptake systems in marine Vibrio and Pseudomonas isolates from the kelp beds off the South African west coast were examined. The fermentative Vibrio isolate possessed a constitutive rapid mannitol uptake system and also a soluble mannitol-1-phosphate dehydrogenase, indicative of a mannitol phosphotransferase system. An inducible, relatively less active mannitol uptake system was detected in the oxidative Pseudomonas isolate, and this strain possessed a mannitol dehydrogenase. The maintenance of these systems during starvation survival was studied. The Vibrio isolate maintained its initial uptake system for approximately 5 weeks of starvation, after which time the uptake system was replaced by one with a higher affinity for mannitol. The mannitol transport system of the Pseudomonas isolate was depressed early in starvation (30 h) but could be readily induced by exogenous mannitol after 6 weeks of starvation. The relative proportions of mannitol which was incorporated and respired were determined in starved Vibrio and Pseudomonas strains. PMID- 16346910 TI - Aerobic Fermentation of D-Xylose to Ethanol by Clavispora sp. AB - Eleven strains of an undescribed species of Clavispora fermented D-xylose directly to ethanol under aerobic conditions. Strain UWO(PS)83-877-1 was grown in a medium containing 2% D-xylose and 0.5% yeast extract, and the following results were obtained: ethanol yield coefficient (ethanol/D-xylose), 0.29 g g (57.4% of theoretical); cell yield coefficient (dry biomass/D-xylose), 0.25 g g; maximum ethanol concentration, 5.9 g liter; maximum volumetric ethanol productivity, 0.11 g liter h. With initial D-xylose concentrations of 40, 60, and 80 g liter, maximum ethanol concentrations of 8.8, 10.9, and 9.8 g liter were obtained, respectively (57.2, 57.1, and 48.3% of theoretical). Ethanol was found to inhibit the fermentation of D-xylose (K(p) = 0.58 g liter) more than the fermentation of glucose (K(p) = 6.5 g liter). The performance of this yeast compared favorably with that reported for some other D-xylose-fermenting yeasts. PMID- 16346911 TI - Habitat segregation and biochemical activities of marine members of the family vibrionaceae. AB - A comparative study of marine members of the family Vibrionaceae with the technique of numerical taxonomy revealed habitat segregation as well as a cosmopolitan nature of species distribution among the vibrios in different marine environments. The bacterial strains analyzed were isolated from seawater, sediments, phyto- and zooplankton, and fish in the Indian Ocean, the South and East China Sea, and West Pacific Ocean, and coastal areas of Japan. A total of 155 morphological, physiological, and biochemical tests were carried out for each of 405 strains examined. The results showed that most of the large taxonomical clusters which emerged from the computation corresponded to ecological groups which have particular niches. For instance, each group of seawater vibrios inhabited a particular water layer of limited depth range, in spite of the fact that strains of the group were isolated from sampling locations spread over a wide area from the Indian Ocean to Japanese coast. Various vibrio groups showed remarkable differences in their physiological and biochemical activities, and the activities of each group seemed to correspond with its ecological niche. The strains which inhabited surface-water layers grew fast and actively utilized many high-molecular-weight organic compounds and carbohydrates that are derived from fresh, easily degradable organic matter present in the surface waters, whereas the middle- and deep-water vibrios did not decompose most of the high-molecular weight organic compounds except chitin but, rather, utilized some carbohydrates and organic acids which seemed to be derived from refractory particulate organic matter present in the deeper waters. PMID- 16346912 TI - Uptake Hydrogenase Activity Determined by Plasmid pRL6JI in Rhizobium leguminosarum Does Not Increase Symbiotic Nitrogen Fixation. AB - Six mutants of Rhizobium leguminosarum 3855 lacking uptake hydrogenase activity (Hup phenotype) as a result of Tn5-mob mutagenesis of the hup-containing plasmid pRL6JI were tested for symbiotic performance on Pisum sativum L. and Vicia benghalensis L. Three pea cultivars and one vetch line, which induce four different levels of Hup activity in strain 3855, were grown to flowering under microbiologically controlled conditions in the absence of combined N. Direct Kjeldahl N measurements showed that in every case at least one Hup mutant fixed as much N(2) as the isogenic Hup strain. Measures of C(2)H(2) reduction, H(2) evolution, H(2) incorporation, and plant dry weight were consistent with the interpretation that the oxidation of H(2) produced by the nitrogenase enzyme complex was not necessarily associated with increased N(2) fixation in these symbiotic associations. Tests with a smaller subset of the Hup strains under four different root environments ranging from pH 5.0 to 8.2 likewise showed no significant advantage for the isogenic Hup strain. It was concluded that the improvements in symbiotic N(2) fixation produced by pRL6JI are associated with some trait other than the Hup phenotype. PMID- 16346913 TI - Denitrification rates in the low-oxygen waters of the stratified baltic proper. AB - Denitrification activity was shown in the deep, low-oxygen waters of the Baltic proper by both in vitro and in situ methods. The vertical distribution of NO(3) in the water column showed nitrate consumption and NO(2) and N(2)O maxima in the deep waters when O(2) was below 0.2 ml liter, which is suggestive evidence for denitrification. Direct in situ evidence for denitrification was obtained by finding an N(2) saturation of up to 108% in the deep waters. When these waters were incubated with NO(3), N(2) was produced. Quantification of the denitrification rate done by the addition of C(2)H(2) to water samples from the active depths showed a rate of about 0.10 mumol liter day. PMID- 16346914 TI - Anaerobic cellulolytic bacteria from wetwood of living trees. AB - Obligately anaerobic, mesophilic, cellulolytic bacteria were isolated from the wetwood of elm and maple trees. The isolation of these bacteria involved inoculation of selective enrichment cultures with increment cores taken from trees showing evidence of wetwood. Cellulolytic bacteria were present in the cores from seven of nine trees sampled, as indicated by the disappearance of cellulose from enrichment cultures. With two exceptions, cellulolytic activity was confined to the darker, wetter, inner section of the cores. Cellulolytic bacteria were also present in the fluid from core holes. The cellulolytic isolates were motile rods that stained gram negative. Endospores were formed by some strains. The physiology of one of the cellulolytic isolates (strain JW2) was studied in detail. Strain JW2 fermented cellobiose, d-glucose, glycerol, l arabinose, d-xylose, and xylan in addition to cellulose. In a defined medium, p aminobenzoic acid and biotin were the only exogenous growth factors required by strain JW2 for the fermentation of cellobiose or cellulose. Acetate and ethanol were the major nongaseous end products of cellulose fermentation. The guanine plus-cytosine content of the DNA of strain JW2 was 33.7 mol%. Cellulolytic bacteria have not previously been reported to occur in wetwood. The isolation of such bacteria indicates that cellulolytic bacteria are inhabitants of wetwood environments and suggests that they may be involved in wetwood development. PMID- 16346915 TI - Degradation of bermuda and orchard grass by species of ruminal bacteria. AB - Fiber degradation in Bermuda grass and orchard grass was evaluated gravimetrically and by scanning and transmission electron microscopy after incubation with pure cultures of rumen bacteria. Lachnospira multiparus D-32 was unable to degrade plant cell wall components. Butyrivibrio fibrisolvens 49 degraded 6 and 14.9% of the fiber components in Bermuda grass and orchard grass, respectively, and Ruminococcus albus 7 degraded 11.4% orchard grass fiber but none in Bermuda grass. Both B. fibrisolvens and R. albus lacked capsules, did not adhere to fiber, and degraded only portions of the more easily available plant cell walls. R. flavefaciens FD-1 was the most active fiber digester, degrading 8.2 and 55.3% of Bermuda and orchard grass fiber, respectively. The microbe had a distinct capsule and adhered to fiber, especially that which is slowly degraded, but was able to cause erosion and disorganization of the more easily digested cell walls, apparently by extracellular enzymes. Results indicated that more digestible cell walls could be partially degraded by enzymes disassociated from cellulolytic and noncellulolytic bacteria, and data were consistent with the hypothesis that the more slowly degraded plant walls required attachment. Microbial species as well as the cell wall architecture influenced the physical association with and digestion of plant fiber. PMID- 16346916 TI - Partial Chemical and Physical Characterization of Two Extracellular Polysaccharides Produced by Marine, Periphytic Pseudomonas sp. Strain NCMB 2021. AB - The marine bacterium Pseudomonas sp. strain NCMB 2021, which can attach to solid, and especially hydrophobic, surfaces, elaborates two different extracellular polysaccharides in batch cultures. One (polysaccharide A) was produced only during exponential growth and contained glucose, galactose, glucuronic acid, and galacturonic acid in a molar ratio of 1.00:0.81:0.42:0.32. It produced viscous solutions, formed gels at high concentrations, and precipitated with several multivalent cations. The other (polysaccharide B) was released at the end of the exponential phase and in the stationary phase. It contained equimolar amounts of N-acetylglucosamine, 2-keto-3-deoxyoctulosonic acid, an unidentified 6 deoxyhexose, and also O-acetyl groups. Despite its high molecular weight (10 to 10 as judged by gel filtration), the polysaccharide produced aqueous solutions with very low viscosities and was also soluble in 90% aqueous phenol, 80% methanol, and 80% ethanol. PMID- 16346917 TI - Purification and Characterization of Liposan, a Bioemulsifier from Candida lipolytica. AB - The inducible water-soluble bioemulsifier liposan (M. C. Cirigliano and G. M. Carman, Appl. Environ. Microbiol. 48:747-750, 1984) was purified from the yeast Candida lipolytica. The purification procedure included repeated solvent extractions of a concentrated culture filtrate and Affi-Gel concanavalin A affinity chromatography. The procedure yielded a preparation containing a major band (M(r) = 27,600) which stained for protein and carbohydrate upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Liposan is composed of approximately 83% carbohydrate and 17% protein. Acid and enzymatic digestions of the emulsifier revealed that the carbohydrate portion is a heteropolysaccharide consisting of glucose, galactose, galactosamine, and galacturonic acid. Liposan effected and stabilized oil-in-water emulsions with a variety of commercial vegetable oils. Emulsification and stabilization properties of liposan were compared to those of a number of commercial emulsifiers and stabilizers. PMID- 16346918 TI - Extracellular Enzymes Produced by the Cultivated Mushroom Lentinus edodes during Degradation of a Lignocellulosic Medium. AB - Although the commercially important mushroom Lentinus (= Lentinula) edodes (Berk.) Sing. can be rapidly cultivated on supplemented wood particles, fruiting is not reliable. This study addressed the problem by developing more information about growth and development on a practical oakwood-oatmeal medium. The study determined (i) the components degraded during a 150-day incubation at 22 degrees C, (ii) the apparent vegetative growth pattern, (iii) the likely growth-limiting nutrient, and (iv) assays that can be used to study key extracellular enzymes. All major components of the medium were degraded, lignin selectively so. The vegetative growth rate was most rapid during the initial 90 days, during which weight loss correlated with glucosamine accumulation (assayed after acid hydrolysis). The rate then slowed; in apparent preparation for fruiting, the cultures rapidly accumulated glucosamine (or its oligomer or polymer). Nitrogen was growth limiting. Certain enzyme activities were associated with the pattern of medium degradation, with growth, or with development. They included cellulolytic system enzymes, hemicellulases, the ligninolytic system, (gluco )amylase, pectinase, acid protease, cell wall lytic enzymes (laminarinase, 1,4 beta-d-glucosidase, beta-N-acetyl-d-glucosaminidase, alpha-d-galactosidase, beta d-mannosidase), acid phosphatase, and laccase. Enzyme activities over the 150-day incubation period with and without a fruiting stimulus are reported. These results provide a basis for future investigations into the physiology and biochemistry of growth and fruiting. PMID- 16346919 TI - Isolation and characterization of a halophilic methanogen from great salt lake. AB - A halophilic methanogenic microorganism isolated from sediments collected from the southern arm of Great Salt Lake, Utah, is described. Cells were irregular, nonmotile cocci approximately 1.0 mum in diameter and stained gram negative. Colonies from anaerobic plates and roll tubes were foamy, circular, and cream yellow. Methanol, methylamine, dimethylamine, and trimethylamine supported growth and methanogenesis. Hydrogen-carbon dioxide, formate, and acetate were not utilized. Sodium and magnesium were required for growth; the optimum NaCl concentration ranged between 1.0 and 2.0 M, with the minimum doubling time occurring at 2.0 M. The optimum growth temperature was 35 degrees C, with maximum growth rate occurring at pH 7.5. The DNA base composition was 48.5 mol% guanine + cytosine. SLP is the type strain designation (= ATCC 35705). PMID- 16346920 TI - Potential for Development of Tolerance by Penicillium digitatum and Penicillium italicum after Repeated Exposure to Potassium Sorbate. AB - Two strains of Penicillium digitatum and one strain of Penicillium italicum were exposed to various levels of sorbic acid and potassium sorbate, and the MICs were determined. Selected strains of the molds were then repeatedly exposed to subinhibitory levels of the compounds to determine whether increased tolerance might develop. The MIC of sorbic acid (pH 4.75) to P. digitatum was between 0.02 and 0.025%. The MIC of sorbate (pH 5.5) to two strains of P. digitatum and P. italicum was found to be between 0.06 and 0.08%. Increasing levels of sorbate resulted in increasing growth suppression of the molds. Populations of P. digitatum were tested for increased tolerance to sorbic acid, and none was found. Individual molds that started from the same parent colony were examined for increased tolerance to potassium sorbate. Two P. digitatum strains developed no observable increased tolerance, but P. italicum developed a slight increase in tolerance to sorbate. When spores of P. italicum and P. digitatum were exposed to higher levels of sorbate for prolonged times, the fungicidal or fungistatic activity of the inhibitor was dependent upon pH, length of exposure time, level of sorbate, and the mold strain. PMID- 16346921 TI - Inhibition experiments on anaerobic methane oxidation. AB - Anaerobic methane oxidation is a general process important in controlling fluxes of methane from anoxic marine sediments. The responsible organism has not been isolated, and little is known about the electron acceptors and substrates involved in the process. Laboratory evidence indicates that sulfate reducers and methanogens are able to oxidize small quantities of methane. Field evidence suggests anaerobic methane oxidation may be linked to sulfate reduction. Experiments with specific inhibitors for sulfate reduction (molybdate), methanogenesis (2-bromoethanesulfonic acid), and acetate utilization (fluoroacetate) were performed on marine sediments from the zone of methane oxidation to determine whether sulfate-reducing bacteria or methanogenic bacteria are responsible for methane oxidation. The inhibition experiment results suggest that methane oxidation in anoxic marine sediments is not directly mediated by sulfate-reducing bacteria or methanogenic bacteria. Our results are consistent with two possibilities: anaerobic methane oxidation may be mediated by an unknown organism or a consortium involving an unknown methane oxidizer and sulfate reducing bacteria. PMID- 16346922 TI - Identification of Glycolytic Enzyme Polypeptides on the Two-Dimensional Protein Map of Saccharomyces cerevisiae and Application to the Study of Some Wine Yeasts. AB - Using a modification of the basic two-dimensional polyacrylamide gel electrophoresis technique, the polypeptides of the protein map of Saccharomyces cerevisiae involved in glycolysis were investigated. This study resulted in a reassignment of two of the seven glycolytic enzyme polypeptides previously identified (Ludwig et al., Mol. Cell. Biol. 2:117-126, 1982), those corresponding to phosphoglycerate kinase and to alcohol dehydrogenase. It also resulted in the identification of two additional glycolytic polypeptides, the enolase B monomer and the glyceraldehyde phosphate dehydrogenase B monomer. The glycolytic enzymes polypeptides so identified were investigated in 5 laboratory strains (all S. cerevisiae) and in 11 commerical strains used for wine making (S. cerevisiae and Saccharomyces bayanus). It appeared highly significant that a particular electrophoretic variant of the glyceraldehyde phosphate dehydrogenase B monomer was found only in the wine yeasts. Furthermore, it was strongly suggested that S. cerevisiae and S. bayanus strains are distinguishible on the basis of a different electrophoretic migration of the enolase B monomer. PMID- 16346923 TI - Microbial degradation of natural rubber vulcanizates. AB - An actinomycete, Nocardia sp. strain 835A, grows well on unvulcanized natural rubber and synthetic isoprene rubber, but not on other types of synthetic rubber. Not only unvulcanized but also various kinds of vulcanized natural rubber products were more or less utilized by the organism as the sole source of carbon and energy. The thin film from a latex glove was rapidly degraded, and the weight loss reached 75% after a 2-week cultivation period. Oligomers with molecular weights from 10 to 10 were accumulated during microbial growth on the latex glove. The partially purified oligomers were examined by infrared and H nuclear magnetic resonance and C nuclear magnetic resonance spectroscopy, and the spectra were those expected of cis-1, 4-polyisoprene with the structure, OHC-CH(2)-[ CH(2)-C(-CH(3))=CH -CH(2)-](n)-CH(2)-C(=O)- CH(3), with average values of n of about 114 and 19 for the two oligomers. PMID- 16346924 TI - Thermophilic anaerobic biodegradation of [C]lignin, [C]cellulose, and [C]lignocellulose preparations. AB - Thermophilic (55 degrees C) anaerobic enrichment cultures were incubated with [C lignin]lignocellulose, [C-polysaccharide]lignocellulose, and kraft [C]lignin prepared from slash pine, Pinus elliottii, and C-labeled preparations of synthetic lignin and purified cellulose. Significant but low percentages (2 to 4%) of synthetic and natural pine lignin were recovered as labeled methane and carbon dioxide during 60-day incubations, whereas much greater percentages (13 to 23%) of kraft lignin were recovered as gaseous end products. Percentages of label recovered from lignin-labeled substrates as dissolved degradation products were approximately equal to percentages recovered as gaseous end products. High pressure liquid chromatographic analyses of CuO oxidation products of sound and degraded pine lignin indicated that no substantial chemical modifications of the remaining lignin polymer, such as demethoxylation and dearomatization, occurred during biodegradation. The polysaccharide components of pine lignocellulose and purified cellulose were relatively rapidly mineralized to methane and carbon dioxide; 31 to 37% of the pine polysaccharides and 56 to 63% of the purified cellulose were recovered as labeled gaseous end products. An additional 10 to 20% of the polysaccharide substrates was recovered as dissolved degradation products. Overall, these results indicate that elevated temperatures can greatly enhance rates of anaerobic degradation of lignin and lignified substrates to methane and low-molecular-weight aromatic compounds. PMID- 16346925 TI - Population Dynamics of Sugar Beets, Rhizoctonia solani, and Laetisaria arvalis: Responses of a Host, Plant Pathogen, and Hyperparasite to Perturbation in the Field. AB - Rhizoctonia solani causes crown rot of sugar beets, a severe disease that has destroyed up to 60% of the plants in a test field in western Nebraska. Laetisaria arvalis, a natural hyperparasite of Rhizoctonia spp., was isolated from fields in western Nebraska. To test for the potential for biological control of R. solani, in November 1980 (following harvest) we applied various combinations of a nematicide (Telone II; Dow Chemical Co.), a nutrition source (sugar beet pulp), and an inoculum of L. arvalis in a randomized block design. Populations of R. solani, L. arvalis, and sugar beets were monitored monthly through October 1981 (just after harvest). In control and nematicide plots, the R. solani population did not change significantly through time. In plots inoculated with L. arvalis, the R. solani populations declined through March, concomitant with an increase in L. arvalis. L. arvalis then declined with a corresponding increase in the R. solani populations. Beet plant numbers declined significantly in all treatments. We suggest that reduction of the R. solani populations with the hyperparasite L. arvalis is possible but that a stable equilibrium naturally exists. PMID- 16346926 TI - Predominance of Fast-Growing Rhizobium japonicum in a Soybean Field in the People's Republic of China. AB - Soybean rhizobia were isolated from two soils with different cropping histories from Hubei province in central China. The first, from Honghu county, has been under soybean cultivation for decades. All of the isolates obtained from nodules on soybeans growing in this soil were fast-growing, acid-producing rhizobia. However, slow-growing, alkali-producing isolates were obtained at higher dilutions of the same soil. The second soil, from Wuchang county, has been under rice cultivation with no record of previous soybean cultivation. All of the soybean rhizobia recovered from this soil, and at higher dilutions of the soil, were typical slow-growing, alkali-producing isolates. The isolates from both soils were grouped by using intrinsic antibiotic resistance, gel immunodiffusion, and fluorescent-antibody procedures. Representative isolates were tested for symbiotic effectiveness with four soybean cultivars (Peking, Davis, Williams, and Ai Jiao Zao) in a pot experiment. There were significant cultivar-rhizobial interactions. Moreover, on each cultivar, there was at least one fast-growing isolate among these new rhizobia that was as effective as the highly effective slow-growing reference strain USDA 110. PMID- 16346927 TI - Photochemical and microbial degradation of 2,4,5-trichloroaniline in a freshwater lake. AB - Surface waters from a eutrophic lake in northern Georgia were incubated with C labeled 2,4,5-trichloroaniline to study the disappearance of the parent compound and production of CO(2). There was no degradation of the compound in the dark. Under 12 h of sunlight and 12 h of darkness, 28% of the trichloroaniline was degraded in both poisoned and untreated samples. Mineralization after 24 h in poisoned and untreated lake water was 5.5 and 6.8%, respectively. Thus, 81% of the mineralization was attributable to photochemical processes, and 19% was attributable to microbial processes. Most biological mineralization was due to microbes of bacterial size (<1.0 mum). Approximately 90% of the trichloroaniline bioaccumulated was associated with organisms larger than 1.0 mum, e.g., algae. When algae were removed by filtration, the amount of trichloroaniline mineralized increased to 9.4%, compared with 6.8% in the presence of algae. The excretion of organic compounds by algae may have inhibited bacterial mineralization of photoproducts. PMID- 16346928 TI - Aerial Dispersal of Epiphytic Bacteria over Bean Plants. AB - Plant canopies are strong sources of bacterial aerosols during sunny days when the leaves are dry. Bacterial concentration, upward flux, and deposition onto exposed petri plates were measured over snap beans during three growing seasons. A net upward flux of bacteria occurred only during the warm part of sunny days, not at night when leaves were wet with dew or when a thermal inversion was present. Aerosol source strength was positively correlated with wind speed. Upward fluxes were higher on days after rain than on days when the soil was dry. Other unidentified sources of variability in source strength probably exist. Canopy-level deposition, apparently due to intermediate-scale transport of bacteria in fairly concentrated clouds, can occur in the early evening. PMID- 16346929 TI - Uncoupling of Methanogenesis from Growth of Methanosarcina barkeri by Phosphate Limitation. AB - Production of methane by Methanosarcina barkeri from H(2)-CO(2) was studied in fed-batch culture under phosphate-limiting conditions. A transition in the kinetics of methanogenesis from an exponentially increasing rate to a constant rate was due to depletion of phosphate from the medium. The period of exponentially increasing rate of methanogenesis was extended by increasing the initial concentration of phosphate in the medium. Addition of phosphate during the constant period changed the kinetics to an exponentially increasing rate of methanogenesis, indicating the reversibility of phosphate depletion. The relation between methanogenesis and growth of M. barkeri was investigated by measuring the incorporation of phosphorus, supplied as KH(2)PO(4), in the medium. At a low (1 muM) initial concentration of phosphate in the medium and during the constant period of methanogenesis, there was no net cell growth. At a higher (10 muM) initial concentration of phosphate, cell growth proceeded linearly with time after phosphate had been removed from the medium by uptake into cells. PMID- 16346930 TI - Interactions in syntrophic associations of endospore-forming, butyrate-degrading bacteria and h(2)-consuming bacteria. AB - Butyrate is an important intermediate in the anaerobic degradation of organic matter. In sulfate-depleted environments butyrate is oxidized to acetate and hydrogen by obligate proton reducers, in syntrophic association with hydrogen consuming methanogens. This paper describes two enrichments of endospore-forming bacteria degrading butyrate in consortia with methanogens. The isolates are readily established in coculture with H(2)-consuming, sulfate-reducing bacteria by pasteurizing the culture. The two original enrichments differed in that one grew to an optically dense culture while the second grew in clumps. Examination by scanning electron microscopy showed that clumping resulted from the production of large amounts of extracellular polymer. Several H(2)-consuming methanogens were identified in the enrichments. Some of them grew closely associated to the butyrate degraders. This attachment to the hydrogen producer may permit some methanogens to compete for the growth substrate against other bacteria having higher substrate affinity. PMID- 16346931 TI - Effect of Oxygen Tension, Mn(II) Concentration, and Temperature on the Microbially Catalyzed Mn(II) Oxidation Rate in a Marine Fjord. AB - We present evidence that the oxidation of Mn(II) in a zone above the O(2)/H(2)S interface in the water column of Saanich Inlet, British Columbia, Canada, is microbially catalyzed. We measured the uptake of Mn(II) in water samples under in situ conditions of pH and temperature and in the presence and absence of oxygen. Experiments in the absence of oxygen provided a measure of the exchange of the tracer between the dissolved and solid pools of Mn(II); we interpret the difference between experiments in the presence and absence of oxygen to be a measure of Mn(II) oxidation. Using this method we examined the effect of oxygen tension, Mn(II) concentration, and temperature on the initial in situ Mn(II) oxidation rate (V(0)). Mn(II) oxidation was almost twice as fast under conditions of 67% air saturation (V(0)=5.5 nM h) as with the in situ concentration of 15 muM (5% air saturation; V(0)=3.1 nM h). Additions of ca. 18 muM Mn(II) completely inhibited all Mn(II) oxidation at three different depths in the oxidizing zone, and there was a temperature optimum for Mn(II) oxidation of around 20 degrees C. These results are consistent with biologically mediated Mn(II) oxidation and indicate that the rate is limited by both oxygen and the concentration of microbial binding sites in this environment. PMID- 16346932 TI - Production of Ligninases and Degradation of Lignin in Agitated Submerged Cultures of Phanerochaete chrysosporium. AB - Research on the extracellular hemeprotein ligninases of Phanerochaete chrysosporium has been hampered by the necessity to produce them in stationary culture. This investigation examined the effects of detergents on development of ligninase activity in agitated submerged cultures. Results show that addition of Tween 80, Tween 20, or 3-[(3-colamidopropyl)dimethylammonio]1-propanesulfonate to the cultures permits development of ligninase activity comparable to that routinely obtained in stationary cultures. The detergent-amended cultures express the entire ligninolytic system, assayed as the complete oxidation of [C]lignin to CO(2). The detergent effect is evidently not merely in facilitating release of extant enzyme. Development of ligninolytic activity in the agitated cultures, as in stationary cultures, is idiophasic. Ion-exchange fast protein-liquid chromatography indicated that the heme protein profiles in agitated and stationary cultures are very similar. These findings should make it possible to scale up production of ligninolytic enzymes in stirred tank fermentors. PMID- 16346934 TI - Influence of leaching parameters on the biological removal of uranium from coal by a filamentous cyanobacterium. AB - Axenic cultures of the filamentous cyanobacterium LPP OL3 were incubated with samples of uraniumbearing coal from a German mining area. The influence of leaching parameters such as coal concentration (pulp density), initial biomass, particle size, temperature, and composition of the growth medium on the leaching of uranium from the ore by the cyanobacterial strain was studied. When low pulp densities were applied, the yield of biologically extracted uranium was optimal (reaching 96% at 1% [wt/vol] coal) and all released uranium was found in the culture liquid. Above 10% (wt/vol) coal in the medium, the amount of cell-bound uranium increased. Initial biomass concentration (protein content of the cultures) and particle size were not critical parameters of leaching by LPP OL3. However, temperature and composition of the growth medium profoundly influenced the leaching of uranium and growth of the cyanobacterium. The yield of leached uranium (at 10% [wt/vol] coal) could not be raised with a tank leaching apparatus. Also, coal ashes were not suitable substrates for the leaching of uranium by LPP OL3. In conclusion, the reactions of the cyanobacterium to variations in leaching parameters were different from reactions of acidic leaching organisms. PMID- 16346933 TI - Effect of fall turnover on terminal carbon metabolism in lake mendota sediments. AB - The carbon and electron flow pathways and the bacterial populations responsible for the transformation of H(2)-CO(2), formate, methanol, methylamine, acetate, ethanol, and lactate were examined in eutrophic sediments collected during summer stratification and fall turnover. The rate of methane formation averaged 1,130 mumol of CH(4) per liter of sediment per day during late-summer stratification versus 433 mumol of CH(4) per liter of sediment per day during the early portion of fall turnover, whereas the rate of sulfate reduction was 280 mumol of sulfate per liter of sediment per day versus 1,840 mumol of sulfate per liter of sediment per day during the same time periods, respectively. The sulfate-reducing population remained constant while the methanogenic population decreased by one to two orders of magnitude during turnover. The acetate concentration increased from 32 to 81 mumol per liter of sediment while the acetate transformation rate constant decreased from 3.22 to 0.70 per h, respectively, during stratification versus turnover. Acetate accounted for nearly 100% of total sedimentary methanogenesis during turnover versus 70% during stratification. The fraction of CO(2) produced from all C-labeled substrates examined was 10 to 40% higher during fall turnover than during stratification. The addition of sulfate, thiosulfate, or sulfur to stratified sediments mimicked fall turnover in that more CO(2) and CH(4) were produced. The addition of Desulfovibrio vulgaris to sulfate-amended sediments greatly enhanced the amount of CO(2) produced from either [C]methanol or [2-C]acetate, suggesting that H(2) consumption by sulfate reducers can alter methanol or acetate transformation by sedimentary methanogens. These data imply that turnover dynamically altered carbon transformation in eutrophic sediments such that sulfate reduction dominated over methanogenesis principally as a consequence of altering hydrogen metabolism. PMID- 16346935 TI - Incidence of Plasmids in Thermus spp. Isolated in Yellowstone National Park. AB - Forty-eight strains of Thermus spp. were isolated from thermal sites in Yellowstone National Park, Wyo., and 62.5% showed evidence of plasmid DNA. Attempts to assign function to the plasmid DNA were unsuccessful, and the presence of plasmid DNA could not be correlated with antibiotic or heavy metal resistance. A number of these cryptic plasmids are now being investigated for their potential as vectors for molecular cloning in Thermus spp. PMID- 16346936 TI - Isolation of Gregatin A from Phialophora gregata by Preparative High-Pressure Liquid Chromatography. AB - A method was developed for the production and purification of gregatin A from Phialophora gregata NRRL 13198 cultured on rice at 20 degrees C for 28 days. Liquid extraction followed by high-pressure liquid chromatography afforded 247.0 mg of crystalline gregatin A per kg of rice. PMID- 16346937 TI - Nutrient-Enhanced Production of Remarkably High Concentrations of Ethanol by Saccharomyces bayanus through Soy Flour Supplementation. AB - The supplementation of a simple medium with soy flour led to an increase in the specific growth rate and viable cell concentration of Saccharomyces bayanus during fermentation. Increasing the amount of soy flour led to an increase in the maximum number of viable yeast cells and the percentage of glucose fermented. It was possible in 64 h to reach 12.8% (wt/vol) ethanol by adding 4% soy flour (wt/vol) to a simple medium with 300 g of glucose per liter. The aqueous extract from soy flour was nearly as effective as whole-soy flour, whereas the lipidic fraction had no positive effect. PMID- 16346938 TI - Diffusion through a Double-Sided Plate: Development of a Method to Study Alga Bacterium Interactions. AB - Bacteria and algae isolated from a wastewater oxidation pond were inoculated onto opposing surfaces of double-layer agar plates (Lutri plates) to determine the usefulness of such plates for studying microbial interactions. The altered growth characteristics of various algae depending on the species of bacteria on the adjacent medium surface indicated that there was diffusion of extracellular products through the agar, suggesting that this simple assay can be used for screening potential interactions of actively growing organisms. PMID- 16346939 TI - Responses of Ruminococcus flavefaciens, a Ruminal Cellulolytic Species, to Nutrient Starvation. AB - Ruminococcus flavefaciens strain C94, a strictly anaerobic, cellulolytic ruminal bacterial species, was grown either in batch or continuous cultures (cellobiose limited or nitrogen limited) at various dilution rates. Washed cell suspensions were incubated anaerobically at 39 degrees C without nutrients for various times up to 24 h. The effects of starvation on direct and viable cell counts, cell composition (DNA, RNA, protein, and carbohydrate), and endogenous production of volatile fatty acids by the cell suspensions were determined. In addition, the effect of the pH of the starvation buffer on direct and viable cell counts was determined. Survival of batch-grown cells during starvation was variable, with an average time for one-half the cells to lose viability (ST(50)) of 10.9 h. We found with continuous cultures that viable cell counts declined faster when the initial cell suspensions had been grown at faster dilution rates; this effect was more pronounced for suspensions that had been limited by cellobiose (ST(50) = 6.6 h at a dilution rate of 0.33 h) than for suspensions that had been limited by nitrogen (ST(50) = 9.5 h at a dilution rate of 0.33 h). With continuous cultures, viable cell counts in all cases declined faster than direct cell counts did. The rates of disappearance of specific cell components during starvation varied with the initial growth conditions, but could not be correlated with the loss of viability. Volatile fatty acid production by starving cells was very low, and acetate was the main product. Starved cells survived longer at pH 7.0 than they did at pH 5.5, and this effect of pH was greater for cellobiose-limited cells (mean ST(50) = 7.1 h) than for nitrogen-limited cells (mean ST(50) = 12 h). Although it has relatively low ST(50) values, R. flavefaciens has sufficient survival abilities to maintain reasonable numbers in domestic animals having maintenance or greater feed intake. PMID- 16346940 TI - Physical and chemical correlates of microbial activity and biomass in composting municipal sewage sludge. AB - Various physical and chemical parameters were monitored to evaluate their influence on the microbial communities present in composting municipal sewage sludge. Temperature, moisture content, depth, pH, protein content, total nitrogen, total carbon, lipid phosphate biomass, and the rates of microbial incorporation of substrates into lipids were measured at several times throughout the 17- to 19-day composting runs. Temperature was found to have the most consistent and dramatic effect on microbial activity and biomass. When temperatures exceeded 55 to 60 degrees C, microbial activity fell dramatically, usually by more than 1 order of magnitude. Microbial activity was generally greatest in samples taken from the 35 to 50 degrees C areas of the composting piles. Changes in the composition of the compost over time included increased pH, increased protein content, and decreased total organic content. The changes in these parameters appeared to reflect the microbial activity and biomass present. The results of this study indicate that the rate of composting may best be optimized by controlling the composting temperatures, provided that the other parameters fall within reasonable limits in the starting material. PMID- 16346941 TI - Effects of Low-Temperature Acclimation and Oxygen Stress on Tocopheron Production in Euglena gracilis Z. AB - The effects of low-temperature acclimation and oxygen stress on tocopheron production were examined in the unicellular phytoflagellate Euglena gracilis Z. Cells were cultured photoheterotrophically at 27.5 +/- 1 degrees C with 5% carbon dioxide-95% air and 740 microeinsteins m s (photosynthetically active radiation) and served as controls. Low-temperature acclimation (12.5 +/- 1 degrees C) and high-oxygen stress (5% carbon dioxide-95% oxygen) were individually examined in the mass culturing of the algae. Chromatographic analyses demonstrated a six-to sevenfold enhancement of alpha-tocopherol production in temperature-stressed cells, along with a concomitant decline in the levels of alpha-tocotrienol and the absence of other tocopherol homologs. Oxygen-stressed cultures demonstrated the presence of high levels of alpha-tocopherylquinone; alpha-tocopheron and its homologs and precursors were absent or declined markedly. These findings are discussed in terms of the feasibility of microbial production of natural tocopherols. In addition, these results lend themselves to speculation regarding the biological role(s) of tocopherols as antioxidants and free radical scavengers in reducing photo-induced oxidative damage or lipid peroxidation toxicities or both in photosynthetically active E. gracilis Z. PMID- 16346942 TI - Characterization of Rhizobia from Ineffective Alfalfa Nodules: Ability to Nodulate Bean Plants [Phaseolus vulgaris (L.) Savi.]. AB - This study was initiated to characterize Rhizobium isolates obtained from root nodules of ineffectively nodulated, field-grown alfalfa (Medicago sativa L.) plants. The purpose was to determine if these isolates possessed characteristics which would explain either their ineffectiveness in N(2) fixation or their apparent ability to tolerate the moderately acid soil conditions from which they originated. Isolates were characterized by analysis of growth rate, 39 degrees C tolerance, acid production on conventional media, and symbiotic performance. All isolates were ineffective in N(2) fixation on alfalfa, and they contained one or more anomalous characteristics. These included either slow growth rate, lack of 39 degrees C tolerance, or lack of acid production on conventional media. Infectiveness tests on a broad range of legumes revealed that the isolates formed root nodules on M. sativa, Medicago lupulina L., and Phaseolus vulgaris (L.) Savi. (common bean). These results provide evidence that, in some situations, ineffective nodulation of M. sativa in the field may be due to the presence of promiscuous, native Rhizobium species. PMID- 16346943 TI - Examination of thermophilic methane-producing digesters by analysis of bacterial lipids. AB - Thermophilic methane-producing digesters were examined by the analysis of lipids to determine the microbial biomass, community structure, and nutritional status of the microbes within the digesters. The digesters received a daily feedstock of cattle feed and Bermuda grass, with some digesters receiving additional supplements of propionate, butyrate, or nitrate. Microbial biomass, measured as total extractable lipid phosphate, was decreased in slurries from digesters receiving continuous addition of the fermentation intermediates propionate or butyrate as compared with slurries from control digesters receiving the feedstock alone. In slurries from digesters that received continuous addition of nitrate, the microbial biomass was higher than in the slurries from control digesters. The control digesters had ca. 2.5 x 10 bacteria per g (dry weight) as determined from total extractable lipid phosphate. Shifts in microbial community structure were observed by analysis of ester-linked phospholipid fatty acids. Statistical analysis of the patterns of phospholipid fatty acids indicated that the digesters receiving different supplements could be distinguished from the control digester and from each other. Poly-beta-hydroxybutyric acid, an indicator of metabolic stress, was detected in slurries from all the digesters. Slurries from the nitrate-amended digester had the highest concentration of poly-beta hydroxybutyric acid, whereas slurries from the propionate-amended digester had the lowest concentration. These chemical analyses offer a quantitative means to correlate shifts in microbial biomass, community structure, and nutritional status in complex fermentation systems to the production of a specific end product. PMID- 16346944 TI - Influence of deep ocean sewage outfalls on the microbial activity of the surrounding sediment. AB - The microbial activity near two deep ocean sewage outfalls off the coast of the island of Oahu, Hawaii, was characterized. Water samples and sediment samples to a depth of 4.5 cm were analyzed from an area of approximately 4.5 x 10 m surrounding the outfalls. Although the effluent water at both sites exhibited heterotrophic activity that was 2 orders of magnitude greater than water from a control site, ambient water samples taken within 1 m of the discharge ports exhibited activity only twice that of the control water. The heterotrophic activity of the outfall sediment was only elevated above that of the control site for surface samples collected within 10 m of the outfall. Likewise, the rates of microbial nucleic acid synthesis and carbon production in the sediment were only elevated immediately adjacent to the outfalls. Total microbial biomass, as determined by the ATP content of the sediment, varied spatially but was generally elevated at the outfall sites. The specific growth rates calculated for the sediment microbial populations, however, were not greater at the outfall sites. At one site the rocks surrounding the diffuser pipe were covered with copious amounts of slime that appeared to be composed entirely of microbial cells and filaments. This microbial mat was extremely active with respect to heterotrophic activity and biomass production. Overall, it appears that the impact of the sewage discharge on the ambient seawater microbiota is slight and that the effect on the sediment microbiota is confined to an area immediately adjacent to the diffuser ports. In the sand itself, the effect is limited to the upper 2 cm at most. PMID- 16346945 TI - Purification and Characterization of Extracellular Amylolytic Enzymes from the Yeast Filobasidium capsuligenum. AB - The extracellular amylolytic system of Filobasidium capsuligenum consisted of an alpha-amylase (1,4-alpha-d-glucan glucanhydrolase, EC 3.2.1.1) and two forms of glucoamylase (1,4-alpha-d-glucan glucohydrolase, EC 3.2.1.3). The enzymes were purified by ammonium sulfate fractionation, repeated ion-exchange chromatography (DEAE-Sephadex A-50), and gel filtration (Sephadex G-25, Sephadex G-100 sf). alpha-Amylase had an optimum pH of 5.6 and an optimum temperature of 50 degrees C but was rapidly inactivated at higher temperature. The molecular weight was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 64,000. An acarbose concentration of 20 mug/ml was required for 50% inhibition of the alpha-amylase. Both glucoamylases are glycoproteins of identical molecular weight (60,000) and produce only glucose by exohydrolysis. The debranching activity of the glucoamylases was evidenced with substrates containing alpha-1,6 linkages. The pH optima were 5.0 to 5.6 for glucoamylase I and 4.8 to 5.3 for glucoamylase II. Glucoamylase I had a higher optimum temperature (55 degrees C) than glucoamylase II (50 degrees C) and was also more resistant to thermal inactivation. Only low acarbose concentrations (<0.1 mug/ml) were required to reduce the activity of the glucoamylases by 50%. PMID- 16346946 TI - Production of High-Viscosity Whey Broths by a Lactose-Utilizing Xanthomonas campestris Strain. AB - Xanthomonas campestris BB-1L was isolated by enrichment and selection by serial passage in a lactose-minimal medium. When BB-1L was subsequently grown in medium containing only 4% whey and 0.05% yeast extract, the lactose was consumed and broth viscosities greater than 500 cps at a 12 s shear rate were produced. Prolonged maintenance in whey resulted in the loss of the ability of BB-1L to produce viscous broths in whey, indicating a reversion to preferential growth on whey protein, like the parent strain. PMID- 16346947 TI - Isolation and Screening of Yeasts That Ferment d-Xylose Directly to Ethanol. AB - Natural habitats of yeasts were examined for the presence of strains able to produce ethanol from d-xylose. Black knots, insect frass, and tree exudates were screened by enrichment in liquid d-xylose-yeast extract medium. These and each d xylose-assimilating yeast in a collection from cactus fruits and Drosophila spp. were tested for alcohol production from this sugar. Among the 412 isolates examined, 36 produced more than 1 g of ethanol liter from 20 g of d-xylose liter, all under aerated conditions. Closer examination of the strains indicated that their time courses of d-xylose fermentation followed different patterns. Some strains produced more biomass than ethanol, and among these, ethanol may or may not be assimilated rapidly after depletion of d-xylose. Others produced more ethanol than biomass, but all catabolized ethanol after carbohydrate exhaustion. Ethanol production appeared best at low pH values and under mild aeration. Possible correlations between the nutritional profiles of the yeasts and their ability to produce ethanol from d-xylose were explored by multivariate analysis. d-Xylose appeared slightly better utilized by yeasts which rate poorly in terms of fermentation. The fermentation of d-glucose had no bearing on d-xylose fermentation. No specific nutritional trait could discriminate well between better d-xylose fermentors and other yeasts. PMID- 16346948 TI - Temporal Occurrence of Vibrio Species and Aeromonas hydrophila in Estuarine Sediments. AB - Marine sediments were assayed for their concentration of Vibrio spp. and Aeromonas hydrophila over 1 year. A temporal variation was observed in which A. hydrophila, and to a lesser degree V. fluvialis, were found in the winter months, V. parahaemolyticus and V. vulnificus predominated in the spring and summer, with non-O-1 V. cholerae and V. alginolyticus detected in the late summer and fall. These organisms were found in greatest numbers in the top 5 cm of sediment, but were detectable down to 15 cm. Epidemiological data revealed a predominance of non-O-1 V. cholerae infections at the time the organisms were observed to flourish in the sediments. PMID- 16346949 TI - Survival of Bacillus thuringiensis Spores in Soil. AB - Bacillus thuringiensis spores and parasporal crystals were incubated in natural soil, both in the laboratory and in nature. During the first 2 weeks, the spore count decreased by approximately 1 log. Thereafter, the number of spore CFU remained constant for at least 8 months. B. thuringiensis did not lose its ability to make the parasporal crystals during its residence in soil. Spore survival was similar for a commercial spore-crystal preparation (the insecticide) and for laboratory-grown spores. In contrast to these results, spores that were produced in situ in soil through multiplication of added vegetative cells survived for only a short time. For spore additions to soil, variations in soil pH had little effect on survival for those spores that survived the first 2 weeks of incubation. Also without effect were various pretreatments of the spores before incubation in soil or nutritional amendment or desiccation of the soil. Remoistening of a desiccated soil, however, caused a decrease in spore numbers. Spores incubated in soil in the field did not show this, but the degree of soil desiccation in nature probably never reached that for the laboratory samples. The good survival of B. thuringiensis spores after the first 2 weeks in soil seemed to be a result of their inability to germinate in soil. We found no evidence for the hypothesis that rapid germination ability for spores in soil conferred a survival advantage. PMID- 16346950 TI - Arylglycerol-gamma-Formyl Ester as an Aromatic Ring Cleavage Product of Nonphenolic beta-O-4 Lignin Substructure Model Compounds Degraded by Coriolus versicolor. AB - 4-Ethoxy-3-methoxyphenylglycerol-gamma-formyl ester (compound IV) was identified as a degradation product of both 4-ethoxy-3-methoxyphenylglycerol-beta syringaldehyde ether (compound I) and 4-ethoxy-3-methoxyphenylglycerol-beta-2,6 dimethoxyphenyl ether (compound II) by a ligninolytic culture of Coriolus versicolor. An isotopic experiment with a C-labeled compound (compound II') indicated that the formyl group of compound IV was derived from the beta-phenoxyl group of beta-O-4 dimer as an aromatic ring cleavage fragment. However, compound IV was not formed from 4-ethoxy-3-methoxyphenylglycerol-beta-guaiacyl ether (compound III). gamma-Formyl arylglycerol (compound IV) could be a precursor of 4 ethoxy-3-methoxyphenylglycerol (compound VI), because 3-(4-ethoxy-3 methoxyphenyl)-1-formyloxy propane (compound VII) was cleaved to give 3-(4-ethoxy 3-methoxyphenyl)-1-propanol (compound VIII) by C. versicolor. 4-Ethoxy-3 methoxyphenylglycerol-beta,gamma-cyclic carbonate (compound V), previously found as a degradation product of compound III by Phanerochaete chrysosporium (T. Umezawa, and T. Higuchi, FEBS Lett., 25:123-126, 1985), was also identified from the cultures with compound I, II, and III and degraded to give the arylglycerol (compound VI). An isotopic experiment with C-labeled compounds II' and III' indicated that the carbonate carbon of compound V was derived from the beta phenoxyl groups of beta-O-4 substructure. PMID- 16346951 TI - Nitrous oxide emission associated with autotrophic ammonium oxidation in Acid coniferous forest soil. AB - Aerobic N(2)O production was studied in nitrifying humus from urea-fertilized pine forest soil. Acetylene and nitrapyrin inhibited both NH(4) oxidation and N(2)O production, indicating that N(2)O production was closely associated with autotrophic NH(4) oxidation. N(2)O production was enhanced by low soil pH; it was negligible above pH 4.7. When soil pH decreased from 4.7 to 4.1, the relative amount of N(2)O-N produced from NH(4)-N oxidized increased exponentially to 20%. There was also some evidence that N(2)O formation was stimulated by salts (potassium sulfate and sodium phosphates). The maximum rate of N(2)O-N production was 0.17 mug of N(2)O-N per g of soil per h. When humus was treated with NO(2), N(2)O evolved immediately, indicating chemical formation, but no N(2)O was formed on the addition of NO(3). The amount of N(2)O-N evolved was 0.6 to 4.6% of NO(2) N added. A high concentration of NO(2) and low soil pH enhanced chemical production of N(2)O. There was no accumulation of NO(2) during nitrification. The calculations indicated that chemical formation of N(2)O was not the main source of N(2)O during NH(4) oxidation. After the addition of inhibitors of NH(4) oxidation the soils contained NO(3), but no N(2)O was produced. The results suggest that enhanced autotrophic NH(4) oxidation is a potential source of N(2)O in fertilized acid forest soil. PMID- 16346952 TI - Effect of temperature on composting of sewage sludge. AB - The effect of temperature on the composting reaction of sewage sludge was investigated at 50, 60, and 70 degrees C. The total amount of CO(2) evolved and the final conversion of volatile matter were maximum at 60 degrees C., suggesting that the optimal temperature for composting was around 60 degrees C. The specific CO(2) evolution rate (moles of CO(2) evolved per hour per viable cell) was maximum at 70 degrees C. The isolated thermophilic bacterium which was dominant at 60 degrees C but did not grow at 70 degrees C showed that the rate of O(2) consumption measured on the agar plate at 70 degrees C was four times higher than that at 60 degrees C. This showed that the energy yielded from catabolism is rather uncoupled with the anabolism at 70 degrees C in the metabolism of microorganisms indigenous in the compost. A higher respiratory quotient was observed at 70 degrees C than at any other temperature. PMID- 16346953 TI - Yellow Blotch of Pleurotus ostreatus. AB - Yellow blotch disease of the oyster mushroom (Pleurotus ostreatus) was first observed in a commercial mushroom farm in California in 1983. The disease, caused by Pseudomonas agarici, is characterized by primordia, with yellow droplets on their surface, which become stunted, yellow to orange, and deformed as they mature. PMID- 16346954 TI - Association of a 13.6-Megadalton Plasmid in Pediococcus pentosaceus with Bacteriocin Activity. AB - Pediococcus pentosaceus FBB61 and L7230, isolated from cucumber fermentations, produced a bacteriocin, designated pediocin A, which had identical activity spectra against species of Pediococcus, Clostridium, Staphylococcus, and Streptococcus. Both strains possessed a 13.6-megadalton plasmid (pMD136). Plasmid curing experiments suggested that both bacteriocin immunity and production determinants were encoded by pMD136. Use of pediocin-producing strains in food fermentations is discussed. PMID- 16346955 TI - Methanogenic bacteria, including an Acid-tolerant strain, from peatlands. AB - Five pure cultures of methanogenic bacteria were isolated from Minnesota peatlands by enrichment culture techniques. One strain, identified as a member of the family Methanobacteriaceae by antigenic fingerprinting, was acid tolerant and able to produce methane at pH 3.1. Growth could not be demonstrated at pH less than 5.3. PMID- 16346956 TI - Interaction of Pseudomonas putida ATCC 12633 and Bacteriophage gh-1 in Berea Sandstone Rock. AB - Measurements of the passage of Pseudomonas putida ATCC 12633 and a phage resistant mutant through Berea sandstone rock were made. When bacteriophage gh-1 was adsorbed within the rock matrix, a reduction in the passage of the susceptible but not the resistant cells through the rock was observed. PMID- 16346957 TI - Selective medium for recovering specific populations of rhizobia introduced into tropical soils. AB - Experiments were designed to evaluate the usefulness of antifungal agents and streptomycin for recovering low densities of rhizobia inoculated into tropical soils. The results showed that yeast-mannitol agar (pH 6.0) containing 500 mug of streptomycin, 400 mug of cycloheximide and 50 mug of benomyl or chlorothalonil per ml was the best selective medium. PMID- 16346958 TI - Evidence for Plasmid Linkage of Raffinose Utilization and Associated alpha Galactosidase and Sucrose Hydrolase Activity in Pediococcus pentosaceus. AB - The ability to ferment the trisaccharide raffinose was linked with the presence of plasmid DNA in three strains of Pediococcus pentosaceus. Parental strains showed associated inducible alpha-galactosidase and sucrose hydrolase activities when grown in alpha-galactosides and sucrose, respectively. Derivative strains of PPE1.0, PPE2.0, and PPE5.0, which had lost 30-, 28-, and 23-megadalton plasmids, respectively, had no alpha-galactosidase or sucrose hydrolase activity. PMID- 16346959 TI - Methods for measuring specific rates of mercury methylation and degradation and their use in determining factors controlling net rates of mercury methylation. AB - A method was developed to estimate specific rates of demethylation of methyl mercury in aquatic samples by measuring the volatile C end products of CH(3)HgI demethylation. This method was used in conjunction with a Hg radiochemical method which determines specific rates of mercury methylation. Together, these methods enabled us to examine some factors controlling the net rate of mercury methylation. The methodologies were field tested, using lake sediment samples from a recently flooded reservoir in the Southern Indian Lake system which had developed a mercury contamination problem in fish. Ratios of the specific rates of methylation/demethylation were calculated. The highest ratios of methylation/demethylation were calculated. The highest ratios of methylation/demethylation occurred in the flooded shorelines of Southern Indian Lake. These results provide an explanation for the observed increases in the methyl mercury concentrations in fish after flooding. PMID- 16346960 TI - Intermediate-scale, semicontinuous solid-phase fermentation process for production of fuel ethanol from sweet sorghum. AB - A novel, semicontinuous solid-phase fermentation system was used to produce fuel ethanol from sweet sorghum. The process was at an intermediate scale. In the process, dried and shredded sweet sorghum was rehydrated to 70% moisture, acidified to pH 2.0 to 3.0, and either pasteurized (12 h at 70 to 80 degrees C) or not pasteurized before spray inoculation with a broth culture of Saccharomyces cerevisiae. Fermented pulp exited the semicontinuous fermentor after a retention time of 72 h and contained approximately 6% (vol/vol) ethanol. Ethanol yields from dry sweet sorghum were 176 to 179 liters/10 kg (85% of theoretical). Production costs for a greatly scaled-up (x1,400) conceptual version of this system were projected by calculation to average $0.47/liter for 95% ethanol. The calculated energy balance (energy output/energy input ratio) was estimated to be 1.05 when pasteurization was included and 1.31 when pasteurization was omitted. In calculating the energy balances, the output energy of the protein feed byproduct and the input energy for growing the sweet sorghum were not considered. A design for the scaled-up plant (farm scale) is provided. PMID- 16346961 TI - Comparison of Cellulolytic Activities in Clostridium thermocellum and Three Thermophilic, Cellulolytic Anaerobes. AB - Avicelase, carboxymethyl cellulase (CMCase), and beta-glucosidase activities have been compared between Clostridium thermocellum and three extremely thermophilic, cellulolytic anaerobes, isolates TP8, TP11, and KT8. The three isolates were all small, gram-negative staining, oval-ended rods which occurred singly and, at exponential phase, in long chains. They were nonflagellated and no spores were visible. The KT8 and TP11 isolates caused clumping of the cellulose during growth. In all four organisms the CMCase activity paralleled cell growth; however, in C. thermocellum and TP8 the avicelase activity did not increase until early stationary phase. Total CMCase activity in C. thermocellum was significantly higher than in the three isolates; however, avicelase activities were much more comparable among the four organisms. C. thermocellum produced higher levels of ethanol, and all four organisms produced similar concentrations of acetate. The amounts of free and bound CMCase and avicelase activities were investigated. In C. thermocellum and TP8 most of the CMCase and avicelase activities were bound to the cellulose in the medium. In contrast, most of the CMCase activity in TP11 and KT8 was free in the culture supernatant; a significant percentage of avicelase activity was also free. The TP8 isolate was also grown on a defined medium with urea as sole nitrogen source and cellulose serving as the carbon source. Under these conditions the pattern of enzyme production was the same as that in the enriched medium, although the level of that production was considerably reduced. PMID- 16346962 TI - Survival of Azotobacter spp. in Dry Soils. AB - Dry soils stored in glass containers in the laboratory and protected from contamination for periods of 22 to 24 years yielded numerous colonies of Azotobacter chroococcum and other members of the family Azotobacteraceae. These results were compared with those reported in 1974, and the findings are uniformly consistent in terms of surviving populations. The data prove that these bacteria remain viable after prolonged periods of dormancy in much the same way as do the endospores of gram-positive bacteria. PMID- 16346963 TI - Effect of nickelous and other metal ions on the inhibition of rumen bacterial metabolism by 3-(3'-isocyanocyclopent-2-enylidene)propionic Acid and related isocyanides. AB - 3-(3'-Isocyanocyclopent-2-enylidene)propionic acid at a concentration of 2 to 5 mug ml inhibited cellulose digestion by a mixed culture of rumen microorganisms and in other experiments inhibited the degradation of timothy hay (Phleum pratense) in a digestibility test. At isocyanide concentrations of 12 mug ml the fermentation activity of rumen fluid, measured by its dehydrogenase activity, was inhibited but not abolished. All of these isocyanide effects were reversed by the incorporation of nickelous ion into the solutions of the systems under study. The activity of 1 mol of isocyanide is reversed by about 1 mol of Ni and, in the case of the cellulose digestion test, by about 1 mol of Co. Of some 15 other ions tested only Pd and possibly chromium reversed the effect of the isocyanide. PMID- 16346964 TI - Activity of an Attached and Free-Living Vibrio sp. as Measured by Thymidine Incorporation, p-Iodonitrotetrazolium Reduction, and ATP/DNA Ratios. AB - Three independent techniques, [H]thymidine incorporation, the reduction rate of p iodonitrotetrazolium violet (INT) to INT formazan normalized to DNA, and the ratio of ATP to DNA, were adapted to measure the activity of attached and unattached estuarine bacteria. In experiments employing the estuarine isolate Vibrio proteolytica, nutrient concentrations were manipulated by varying the concentration of peptone-yeast extract. In the presence of exogenous nutrients, the activity of free-living cells was greater than that of attached cells as measured by [H]thymidine incorporation and ATP/DNA ratios. In the absence of peptone-yeast extract, however, the activity of attached cells surpassed that of free-living cells as determined by [H]thymidine incorporation and INT formazan normalized to DNA. Of the three techniques, [H]thymidine incorporation was deemed most sensitive for detecting changes in activity resulting from slight differences in nutrient concentration. By this technique, attached cells were much less sensitive to changing nutrient concentrations than were free-living cells. Below a threshold concentration, attached cell activity remained constant, while the activity of unattached cells decreased as a function of decreasing nutrient concentration. The results suggest that loss of cell surface area available for substrate uptake due to attachment may be an important factor in determining the relative activities of attached and free-living cells. PMID- 16346965 TI - Activity measurements of planktonic microbial and microfouling communities in a eutrophic estuary. AB - [H]thymidine incorporation, the rate of reduction of iodonitrotetrazolium violet (INT) to INT formazan normalized to DNA, and the ratio of ATP to DNA were adapted to measure the activity of attached and unattached microbial assemblages of Bayboro Harbor, Fla. Activity measurements by [H]thymidine incorporation were made of cells attached to polystyrene culture dishes, in unfiltered water samples, and in the <1-mum-filtered fraction. In most cases, the activity of attached cells was greater than that of unattached cells either in unfiltered water samples or in the <1-mum fraction. The calculated thymidine incorporation rates for cells in the >1-mum fraction were higher than those for cells either in unfiltered water or in the <1-mum-filtered fraction. By the rate of reduction of INT to INT formazan normalized to DNA and by ATP-to-DNA ratios, attached cells were also more active than cells in unfiltered water samples. These results indicate that the microenvironment afforded by attachment is a more beneficial habitat for microbial growth. Reasons for greater activity by natural populations of attached bacteria are discussed. PMID- 16346966 TI - Metabolism and metal binding by surface-colonizing bacteria: results of microgradient measurements. AB - Short-term (65-h) bacterial colonization of 0.2-mum (pore size) filters submerged in water from Lake Charlotte, Nova Scotia, was characterized by a well-defined succession of cell types, in which small cocci gave way to larger, rod-shaped cells. This succession agrees with the concept of attachment as a strategy for survival, in which inactive cocci can attach to a surface and grow into larger, rod-shaped cells by using endogenous nutrients and the nutrients accumulated at the solid-liquid interface. Analyses of oxygen and CO(2) microgradients above colonized surfaces indicated that a peak of respiration accompanied the succession of rods from cocci. CO(2) fixation then became apparent as the rods began to bind manganese and iron to their surfaces. This means that survival by attachment may not be just the province of heterotrophs. It could also be a strategy adopted by metal-oxidizing chemoautotrophs. Long-term (34-day) colonization of similar filters indicated that, while a succession of attached cell types may indeed be a natural occurrence, other factors (such as the selective grazing of larger cells) tend to obscure the development of this succession. PMID- 16346967 TI - Catabolic Fate of Streptomyces viridosporus T7A-Produced, Acid-Precipitable Polymeric Lignin upon Incubation with Ligninolytic Streptomyces Species and Phanerochaete chrysosporium. AB - Degradation of ground and hot-water-extracted corn stover (Zea mays) lignocellulose by Streptomyces viridosporus T7A generates a water-soluble lignin degradation intermediate termed acid-precipitable polymeric lignin (APPL). The further catabolism of T7A-APPL by S. viridosporus T7A, S. badius 252, and S. setonii 75Vi2 was followed for 3 weeks in aerated shake flask cultures at 37 degrees C in a yeast extract-glucose medium containing 0.05% (wt/vol) T7A-APPL. APPL catabolism by Phanerochaete chrysosporium was followed in stationary cultures in a low-nitrogen medium containing 1% (wt/vol) glucose and 0.05% (wt/vol) T7A-APPL. Metabolism of the APPL was followed by turbidometric assay (600 nm) and by direct measurement of APPL recoverable from the medium. Accumulation and disappearance of soluble low-molecular-weight products of APPL catabolism were followed by gas-liquid chromatography and by high-pressure liquid chromatography, utilizing a diode array detector. Identified and quantified compounds present in culture media included p-coumaric acid, ferulic acid, p hydroxybenzoic acid, p-hydroxybenzaldehyde, protocatechuic acid, vanillic acid, and vanillin. The further catabolism of these APPL-derived aromatic compounds varied with the culture examined, and only S. setonii and P. chrysosporium completely degraded all of them. Some new intermediates of APPL metabolism also appeared in culture media, but the patterns were culture specific. Additional evidence from high-pressure liquid chromatography analyses indicated that one strain, S. badius, converted a water-soluble fraction evident by high-pressure liquid chromatography (7 to 10 min retention time range) into new products appearing at shorter retention times. Mineralization of a [C-lignin]APPL was also followed. The percent C recovered as CO(2), C-APPL, C-labeled water-soluble products, and cell mass-associated radioactivity, were determined for each microorganism after 1 and 3 weeks of incubation in bubbler tube cultures at 37 degrees C. P. chrysosporium evolved the most CO(2) (10%), and S. viridosporus gave the greatest decrease in recoverable C-APPL (23%). The results show that S. badius was not able to significantly degrade the APPL, while the other microorganisms demonstrated various APPL-degrading abilities. The significance of these findings relative to the fate of APPLs in nature was discussed. PMID- 16346968 TI - Measurement of nitrous oxide reductase activity in aquatic sediments. AB - Denitrification in aquatic sediments was measured by an N(2)O reductase assay. Sediments consumed small added quantities of N(2)O over short periods (a few hours). In experiments with sediment slurries, N(2)O reductase activity was inhibited by O(2), C(2)H(2), heat treatment, and by high levels of nitrate (1 mM) or sulfide (10 mM). However, ambient levels of nitrate (<100 muM) did not influence activity, and moderate levels (about 150 muM) induced only a short lag before reductase activity began. Moderate levels of sulfide (<1 mM) had no effect on N(2)O reductase activity. Nitrous oxide reductase displayed Michaelis-Menten kinetics in sediments from freshwater (K(m) = 2.17 muM), estuarine (K(m) = 14.5 muM), and alkaline-saline (K(m) = 501 muM) environments. An in situ assay was devised in which a solution of N(2)O was injected into sealed glass cores containing intact sediment. Two estimates of net rates of denitrification in San Francisco Bay under approximated in situ conditions were 0.009 and 0.041 mmol of N(2)O per m per h. Addition of chlorate to inhibit denitrification in these intact-core experiments (to estimate gross rates of N(2)O consumption) resulted in approximately a 14% upward revision of estimates of net rates. These results were comparable to an in situ estimate of 0.022 mmol of N(2)O per m per h made with the acetylene block assay. PMID- 16346969 TI - Microbial ecophysiology of whey biomethanation: intermediary metabolism of lactose degradation in continuous culture. AB - The intermediary carbon and electron flow routes for lactose degradation during whey biomethanation were studied in continuous culture. The chemostat was operated under lactose-limited conditions with a 100-h retention time. The carbon balance observed for lactose degradation was 4.65 mmol of CH(4), 4.36 mmol of CO(2) and 1.15 mmol of cellular carbon per mmol of lactose consumed, with other intermediary metabolites (i.e., acetate, lactate, etc.) accounting for less than 2% of the lactose consumed. The carbon and electron recoveries for this biomethanation were 87 and 90%, respectively. C tracer studies demonstrated that lactose biomethanation occurred in three distinct but simultaneous phases. Lactose was metabolized primarily into lactate, ethanol, acetate, formate, and carbon dioxide. During this hydrolytic phase, 82% of the lactose was transformed into lactate. These metabolites were transformed into acetate and H(2)-CO(2) in a second, acetogenic, phase. Finally, the direct methane precursors were transformed during the methanogenic phase, with acetate accounting for 81% of the methane formed. A general scheme is proposed for the exact carbon and electron flow route during lactose biomethanation, which predicts the prevalent microbial populations in this ecosystem. PMID- 16346970 TI - Microbial ecophysiology of whey biomethanation: characterization of bacterial trophic populations and prevalent species in continuous culture. AB - The organization and species composition of bacterial trophic groups associated with lactose biomethanation were investigated in a whey-processing chemostat by enumeration, isolation, and general characterization studies. The bacteria were spatially organized as free-living forms and as self-immobilized forms appearing in flocs. Three dominant bacterial trophic group populations were present (in most probable number per milliliter) whose species numbers varied with the substrate consumed: hydrolytic, 10; acetogenic, 10 to 10; and methanogenic, 10 to 10. The three prevalent species utilizing lactose were identified as Leuconostoc mesenteroides, Klebsiella oxytoca, and Clostridium butyricum. Clostridium propionicum and Desulfovibrio vulgaris were the dominant lactate-consuming, hydrogen-producing acetogenic bacteria, while D. vulgaris was the only significant ethanol-degrading species. Methanosarcina barkeri and Methanothrix soehngenii were identified as the dominant acetate-utilizing methanogens, and Methanobacterium formicicum was the prevalent hydrogen-utilizing methanogen. A microbial food chain is proposed for lactose biomethanation that comprises multiple species in three different groups, with the major hydrogen-producing acetogen being a sulfate-reducing species, D. vulgaris, which functioned in the absence of significant levels of environmental sulfate. PMID- 16346971 TI - Production of High-Viscosity Whey-Glucose Broths by a Xanthomonas campestris Strain. AB - Crude broth viscosities, as high as 700 centipoise at a 12-s shear rate, were produced by fermenting a whey-glucose medium with a Xanthomonas campestris strain. PMID- 16346972 TI - Accumulation of Poly (beta-Hydroxybutyrate) by Halobacteria. AB - Some species of extremely halophilic archaebacteria, Halobacteriaceae, have been shown to accumulate large amounts of poly (beta-hydroxybutyrate) under conditions of nitrogen limitation and abundant carbon source. The production of poly (beta hydroxybutyrate), at least in large quantities, was restricted to two carbohydrate-utilizing species, Halobacterium mediterranei and H. volcanii. In addition to the nutrients in the media, the salt concentration also influenced poly (beta-hydroxybutyrate) accumulation, which was greater at lower salt concentrations. The possible application of these microorganisms for the production of biodegradable plastics is discussed. PMID- 16346973 TI - Some Effects of Diflubenzuron on Growth and Sporogenesis in Streptomyces spp. AB - Diflubenzuron, an insect growth regulator that blocks chitin deposition in insect cuticles, was tested for its effects on morphogenesis of Streptomyces spp. Use of diflubenzuron resulted in reduced dominance of spore hairs, reduced the width of the outer wall, and prevented formation of the inner spore wall in S. bambergiensis. In S. coelicolor, diflubenzuron altered the structure of the fibrillar pattern of spore envelopes. Exposure to diflubenzuron resulted in small increases in exported protein and in a ca. 20% increase in chitinase in both Streptomyces spp. PMID- 16346974 TI - Effect of Sterilization by Dry Heat or Autoclaving on Bacterial Penetration through Berea Sandstone. AB - A study was undertaken to determine why bacteria could penetrate lengths of consolidated sandstone (Berea) faster when the sandstone was sterilized by autoclaving than when dry heat (150 degrees C, 3 h) was used. Changes in permeability, porosity, and pore entrance size of the rock as a result of autoclaving were not sufficient to explain the differences in penetration times observed, but electron dispersion spectroscopy and electron microscopy of the rock revealed changes in mineral composition and clay morphology. Autoclaved cores contained more chloride than dry-heated cores, and the clays of autoclaved cores were aggregated and irregularly shaped. Therefore, the decreases in bacterial penetration rates caused by autoclave sterilization were probably the result of a change in surface charge of the pores of the rock and of a reduction in surface area of clays available for adhesion. The results implied that dry heat sterilization was preferable to autoclaving when examining biotic and abiotic interactions in a native-state rock model. PMID- 16346975 TI - Large plasmids from soil bacteria enriched on halogenated alkanoic acids. AB - Four Pseudomonas species and two Alcaligenes species were isolated from soil with a capacity to grow on halogenated alkanoic acids. They were shown to contain one of five large plasmids. The plasmids had molecular weights ranging from 98,800 to 190,000. They were associated with the ability to utilize the halogenated substrates 2-monochloropropionic acid and 2-monochloroacetic acid and with resistance towards one or more of the heavy metals mercury, selenium, and tellurium. The largest plasmid, pUU204, was shown to be unstable in continuous flow culture when the organism was supplied with succinate as the sole carbon source. The dehalogenase gene associated with pUU204 appeared to be readily transferred to an incP group plasmid, R68-45. PMID- 16346976 TI - Transposon Mutagenesis and Excision of R' Plasmids by Conjugative, Chimeric Plasmid pUW942 in Extra-Slow-Growing Rhizobium japonicum Strains. AB - Transposons Tn501 (specifying mercury resistance) and Tn7 (specifying resistance to trimethoprim and streptomycin) were introduced into extra-slow-growing Rhizobium japonicum by conjugal transfer of the 82 kilobase chimeric plasmid pUW942. Mercury-resistant transconjugants were obtained at a frequency of 10 to 10. The transfer frequency of streptomycin resistance was lower than that of mercury resistance, and Tn7 was relatively unstable. pUW942 was not maintained as an autonomously replicating plasmid in R. japonicum strains. However, some of the Hg transconjugants from the RJ19FY, RJ17W, and RJ12S strains acquired antibiotic markers of the vector plasmid pUW942. Southern hybridization of plasmid and chromosomal DNA of R. japonicum strains with P-labeled pUW942 and pAS8Rep-1, the same plasmid as pUW942 except that it does not contain Tn501, revealed the formation of cointegrates between pUW942 and the chromosome of R. japonicum. More transconjugants with only Tn501 insertions in plasmids or the chromosome were obtained in crosses with strains RJ19FY and RJ17W than with RJ12S. These retained stable Hg both in plant nodules and under nonselective in vitro growth conditions. One of the RJ19FY and two of the RJ12S Hg transconjugants with vector plasmid-chromosome cointegrates conjugally transferred plasmids of 82, 84 or 86, and 90 kilobases, respectively, into plasmidless Escherichia coli C. These plasmids strongly hybridized to pUW942 and EcoRI digests of total DNA of each respective R. japonicum strain but not to indigenous plasmid DNA of the R. japonicum strains. These R' plasmids consisted of pUW942-specific EcoRI fragments and an additional one or two new fragments derived from the R. japonicum chromosome. PMID- 16346977 TI - Enzymatic aryl-o-methyl-C labeling of model lignin monomers. AB - Aryl-O-methyl ethers are abundant in aerobic and anaerobic environments. In particular, lignin is composed of units of this type. Lignin monomers specifically radiolabeled in methoxy, side chain, and ring carbons have been synthesized by chemical procedures and are important in studies of lignin synthesis and degradation, humus formation, and microbial O-demethylation. In this paper attention is drawn to an enzymatic procedure for preparing O-methyl-C labeled aromatic lignin monomers which has not previously been exploited in microbial ecology and physiology studies and which has several advantages compared with chemical synthesis procedures. O-[methyl-C]vanillic and O-[methyl C]ferulic acids were prepared with S-[methyl-C]adenosyl-l-methionine as the methyl donor, using commercially obtained porcine liver catechol-O methyltransferase (EC 2.1.1.6). The specific activity of the methylated products was the same as that of the methyl donor, a maximum of about 58 muCi/mumol, and the yields were 42% (vanillate) and 35% (ferulate). Thus lignin monomers are readily prepared as O-methylated products of the catechol-O-methyltransferase reaction and, with this enzyme method of preparation, would be more widely available than labeled compounds which require chemical synthesis. PMID- 16346978 TI - Anaerobic c(1) metabolism of the o-methyl-C-labeled substituent of vanillate. AB - The O-methyl substituents of aromatic compounds constitute a C(1) growth substrate for a number of taxonomically diverse anaerobic acetogens. In this study, strain TH-001, an O-demethylating obligate anaerobe, was chosen to represent this physiological group, and the carbon flow when cells were grown on O-methyl substituents as a C(1) substrate was determined by C radiotracer techniques. O-[methyl-C]vanillate (4-hydroxy-3-methoxy-benzoate) was used as the labeled C(1) substrate. The data showed that for every O-methyl carbon converted to [C]acetate, two were oxidized to CO(2). Quantitation of the carbon recovered in the two products, acetate and CO(2), indicated that acetate was formed in part by the fixation of unlabeled CO(2). The specific activity of C in acetate was 70% of that in the O-methyl substrate, suggesting that only one carbon of acetate was derived from the O-methyl group. Thus, it is postulated that the carboxyl carbon of the product acetate is derived from CO(2) and the methyl carbon is derived from the O-methyl substituent of vanillate. The metabolism of O-[methyl C]vanillate by strain TH-001 can be described as follows: 3CH(3)OC(7)H(5)O(3) + CO(2) + 4H(2)O --> CH(3)COOH + 2CO(2) + 10H + 10e + 3HOC(7)H(5)O(3). PMID- 16346979 TI - Energetics of Leucyl-Leucine Hydrolysis in Streptococcus cremoris Wg(2). AB - The hydrolysis of the dipeptide leucyl-leucine by whole cells of Streptococcus cremoris Wg(2) was dependent on the presence of the energy source lactose. Incubation of cells with uncouplers or ATPase inhibitors prevented the increase of peptidase activity upon the addition of lactose. Incubation with the ionophore nigericin resulted in decreased peptide hydrolysis activity, while incubation with valinomycin led to increased hydrolysis activity. In the presence of nigericin the DeltapH component of the proton motive force was decreased, while the electrical potential was increased. With valinomycin, the electrical potential was collapsed and the DeltapH was increased. When the external pH was decreased from 8 to 5, the rate of peptide hydrolyzing activity by whole cells increased with increasing DeltapH component. In contrast, the peptide hydrolyzing activity in the cell extract decreased with decreasing external pH. These results indicate that the DeltapH component of the proton motive force determines the leucyl-leucine hydrolyzing activity in S. cremoris Wg(2). PMID- 16346980 TI - Kinetic concepts for measuring microbial rate constants: effects of nutrients on rate constants. AB - We investigated the effect of preincubation of environmental waters amended with inorganic nutrients (nitrogen, phosphorus, and traces of iron and magnesium) on the kinetics of the microbial transformation of phenol, propanil, propyl ester of (2,4-dichlorophenoxy)acetic acid, methyl parathion, Ronnel, and methoxychlor in pond and river waters. No effect on the second-order rate constants for these compounds was observed, although there was an increase in the bacterial populations and the pseudo-first-order rate constants. The use of nutrient amended waters could be a useful tool for estimating second-order rate constants for an expanded number of compounds. This technique would provide a larger data base for predicting the behavior of xenobiotic compounds in the environment by using currently available mathematical models. PMID- 16346981 TI - Solid Medium for Culturing Black Smoker Bacteria at Temperatures to 120 degrees C. AB - A solid, highly thermostable medium, based on the new gelling agent GELRITE, was devised to facilitate the culturing of extremely thermophilic microorganisms from submarine hydrothermal vents. The medium remained solid at temperatures to 120 degrees C at vapor pressures and hydrostatic pressures to 265 atm. It proved useful to its maximum tested limits in isolating colonies of black smoker bacteria from hydrothermal fluids recently collected at the Juan de Fuca Ridge in the Pacific Ocean. PMID- 16346982 TI - Nitrification rates in the baltic sea: comparison of three isotope techniques. AB - Simultaneous measurements of nitrification in the Baltic Sea were made at 10- to 30-m intervals in the months of June and November by three isotope techniques: [N]nitrate dilution, N-serve sensitive [C]bicarbonate incorporation, and [N]ammonium oxidation to nitrite and nitrate. Nitrification rates of 1 to 280 nmol liter day were recorded, and each method showed that the highest rates of nitrification occurred below the halocline. Even in the presence of ammonium, dark incubations of mixed layer (above ca. 50 m) waters never yielded nitrification rates exceeding 45 nmol liter day. The rates measured by the ammonium oxidation method were two- to sevenfold greater than those obtained by C incorporation or N dilution. The merits of each technique are discussed, and it is suggested that the [N]ammonium oxidation method should be used in conjunction with the [C]bicarbonate incorporation method. PMID- 16346983 TI - Nitrogen-fixing pseudomonads isolated from roots of plants grown in the canadian high arctic. AB - Root-associated bacteria capable of reducing acetylene to ethylene (biological nitrogen fixation) were isolated from various native plants grown in the Canadian High Arctic. All the strains belonged to the genus Pseudomonas but varied in several physiological characteristics. The rates of acetylene reduction at 14 or 20 degrees C were higher than at 25 or 9 degrees C. Six strains reduced acetylene at 4 degrees C. All the strains exhibited chemotaxis to l-asparagine in semisolid agar at 4 to 25 degrees C. Eleven strains colonized roots of canola (Brassica campestris cv. Tobin) in field soil at population densities of log 4.3 to log 5.1 CFU/g of fresh root at 14 degrees C and log 4.0 to log 5.2 CFU/g of fresh root at 25 degrees C. Some of these nitrogen-fixing pseudomonad strains demonstrated a competitive advantage for root colonization over other rhizosphere bacteria at low temperatures. The combined capabilities of nitrogen fixation and root colonization by diazotrophic pseudomonads may be useful for the development of a biofertilizer inoculant for temperate and cold regions. PMID- 16346984 TI - Semicontinuous and Continuous Production of Chloroperoxidase by Caldariomyces fumago Immobilized in k-Carrageenan. AB - Three strains of Caldariomyces fumago were immobilized in 4% k-carrageenan and tested for semicontinuous production of chloroperoxidase (CPO). Over an 80-day period, growing in defined medium, C. fumago strains CMI 89362 and ATCC 11925 produced enzyme concentrations of 99 and 71 mg/liter, respectively, during six production periods of 12 to 14 days, while C. fumago DAOM 137632 produced only 24 mg of CPO per liter during six growth periods of 10 days. CPO production was unaffected by various regimens of washing between transfers. Mycelial growth was primarily restricted to the head surface, and bead size increased linearly with time. Attempts to restrict growth but maintain CPO production were unsuccessful. Pigment production, fructose utilization, and pH change in the immobilized cell cultures compared closely with the growth characteristics of free cell cultures. By using an airlift tower fermentor with an external loop run with continuous medium replacement of 20 ml/h (D = 0.016), strain CMI 89362 in bead form produced CPO at 40 mg/liter for 11 days. PMID- 16346985 TI - Quantitative influences of butyrate or propionate on thermophilic production of methane from biomass. AB - Sodium butyrate and sodium propionate were continuously infused into separate 4 liter thermophilic digesters. These digesters were operated at 55 degrees C, had a retention time of 20 days, and had a pH of 7.8. Infusion rates were started at 10 mM day and were increased incrementally when new stable external organic acid pool sizes and new stable gas production rates were observed. Stable conditions were obtained in both digesters at an infusion rate of 15 mM day, with methanogenesis elevated over that of control digesters. Calculations based on expected CH(4) at this infusion rate and measured CH(4) production in the treated and control digesters, however, showed an approximately 25% inhibition of methanogenesis in both digesters. A digester infused with sodium chloride showed little or no inhibition at this infusion rate, but was totally inhibited when its infusion rate was increased to 20 mM day, and cumulative added NaCl reached 0.38 M. The butyrate and propionate-amended digesters tolerated addition rates of 20 mM day, but both failed when they were increased to 25 mM day. These results indicate that the thermophilic digesters could function stably at higher external pool sizes of butyrate or propionate than routinely observed. PMID- 16346986 TI - Influence of macrophyte decomposition on growth rate and community structure of okefenokee swamp bacterioplankton. AB - Dissolved substances released during decomposition of the white water lily (Nymphaea odorata) can alter the growth rate of Okefenokee Swamp bacterioplankton. In microcosm experiments dissolved compounds released from senescent Nymphaea leaves caused a transient reduction in the abundance and activity of water column bacterioplankton, followed by a period of intense bacterial growth. Rates of [H]thymidine incorporation and turnover of dissolved d glucose were depressed by over 85%, 3 h after the addition of Nymphaea leachates to microcosms containing Okefenokee Swamp water. Bacterial activity subsequently recovered; after 20 h [H]thymidine incorporation in leachate-treated microcosms was 10-fold greater than that in control microcosms. The recovery of activity was due to a shift in the composition of the bacterial population toward resistance to the inhibitory compounds present in Nymphaea leachates. Inhibitory compounds released during the decomposition of aquatic macrophytes thus act as selective agents which alter the community structure of the bacterial population with respect to leachate resistance. Soluble compounds derived from macrophyte decomposition influence the rate of bacterial secondary production and the availability of microbial biomass to microconsumers. PMID- 16346987 TI - Purification and Characterization of Benzonitrilases from Arthrobacter sp. Strain J-1. AB - We found two kinds of benzonitrilases, designated benzonitrilases A and B, in a cell extract of Arthrobacter sp. strain J-1 grown on benzonitrile as a sole carbon and nitrogen source. Benzonitrilases A and B were purified approximately 409-fold and 38-fold, respectively. Purified benzonitrilase A appeared to be homogeneous according to the criteria of polyacrylamide gel electrophoresis. Both the enzymes hydrolyzed benzonitrile to benzoic acid and ammonia without forming benzamide as an intermediate. The molecular weights of benzonitrilases A and B were found to be 30,000 and 23,000, respectively. The subunit molecular weight of benzonitrilase A was the same as its molecular weight. The isoelectric points of benzonitrilases A and B were 4.95 and 4.80, respectively. The optimum temperature and pH, respectively, for benzonitrilase A were 40 degrees C and 8.5, and those for benzonitrilase B were 30 degrees C and 7.5. The K(m) values for benzonitrilases A and B were 6.7 mM and 4.5 mM, respectively. Both the enzymes degraded p-tolunitrile, 4-cyanopyridine, and p-chlorobenzonitrile, but they did not attack aliphatic nitriles or amides. Both the enzymes were inhibited by thiol reagents. PMID- 16346988 TI - Generation and Characterization of Tn5 Insertion Mutations in Pseudomonas syringae pv. tomato. AB - Tn5-induced insertion mutations were generated in the Pseudomonas syringae pv. tomato genome by mating this plant pathogen with an Escherichia coli strain carrying the suicide plasmid vector for Tn5, pGS9. Km transconjugants occurred at frequencies ranging from 2 x 10 to 9 x 10; approximately 5.5% of these transconjugants were also Cm, indicating the presence of additional pGS9 DNA sequences. Approximately 1% of the Km Cm mutants were auxotrophic. Southern blot analysis revealed that the Tn5 element had inserted into one unique site on the chromosome for each Km Cm transconjugant examined. Physical and genetic tests of Tn5-induced auxotrophs showed that Tn5 mutations in P. syringae pv. tomato were very stable and that secondary transposition of Tn5 or its insertion sequence IS50 was a rare event. Nine of 920 Km Cm transconjugants screened on tomato seedlings either were avirulent or produced very mild symptoms. Each of the virulence mutants was the result of a unique single-site Tn5 insertion. Five mutants also failed to induce a hypersensitivity reaction on tobacco. PMID- 16346989 TI - Lectin Binding to the Root and Root Hair Tips of the Tropical Legume Macroptilium atropurpureum Urb. AB - Ten fluorescein isothiocyanate-labeled lectins were tested on the roots of the tropical legume Macroptilium atropurpureum Urb. Four of these (concanavalin A, peanut agglutinin, Ricinis communis agglutinin I [RCA-I], wheat germ agglutinin) were found to bind to the exterior of root cap cells, the root cap slime, and the channels between epidermal cells in the root elongation zone. One of these lectins, RCA-I, bound to the root hair tips in the mature and emerging hair zones and also to sites at which root hairs were only just emerging. There was no RCA-I binding to immature trichoblasts. Preincubation of these lectins with their hapten sugars eliminated all types of root cell binding. By using a microinoculation technique, preincubation of the root surface with RCA-I lectin was found to inhibit infection and nodulation by Rhizobium spp. Preincubation of the root surface with the RCA-I hapten beta-d-galactose or a mixture of RCA-I lectin and its hapten failed to inhibit nodulation. Application of RCA-I lectin to the root surface caused no apparent detrimental effects to the root hair cells and did not prevent the growth of root hairs. The lectin did not prevent Rhizobium sp. motility or viability even after 24 h of incubation. It was concluded that the RCA-I lectin-specific sugar beta-d-galactose may be involved in the recognition or early infection stages, or both, in the Rhizobium sp. infection of M. atropurpureum. PMID- 16346990 TI - Characterization of Pseudomonas maltophilia Isolates from Fleece Rot. AB - A total of 286 isolates of Pseudomonas maltophilia was collected from sheep exhibiting brown or yellowish fleece rot and from fly-strike lesions. Enzyme activities for 10 of the isolates were examined by plate tests and with the API ZYM system and compared with the enzymatic profile of a human type strain of P. maltophilia. The fact that ovine isolates of P. maltophilia are biochemically similar to pathogenic human strains suggests there may be an association between this organism and the brown to yellow type of fleece rot. PMID- 16346991 TI - Adaptation of aquatic microbial communities to quaternary ammonium compounds. AB - The effects of long-chain (C(12) to C(18)) quaternary ammonium compounds (QACs) on the density, heterotrophic activity, and biodegradation capabilities of heterotrophic bacteria were examined in situ in a lake ecosystem. Monoalkyl and dialkyl substituted QACs were tested over a range of concentrations (0.001 to 10 mg/liter) in both acute (3 h) and chronic (21 day) exposures. In general, none of the QACs tested had significant adverse effects on bacterial densities in either acute or chronic studies. However, significant decreases in bacterial heterotrophic activity were noted in acute studies at QAC concentrations from 0.1 to 10 mg/liter. Chronic exposure of lake microbial communities to a specific monoalkyl QAC resulted in an adaptive response and recovery of heterotrophic activity. No-observable-effect level in the adapted populations was >10 mg/liter. Chronic exposure also resulted in significant increases in the number and activity of bacteria capable of biodegrading the material. The increase in biodegradation capability was observed at low (microgram per liter) concentrations which are approximately the same as realistic environmental levels. In general, our studies indicated that exposure of lake microbial communities to QACs results in the development of adapted communities which are less sensitive to potential toxic effects and more active in the biodegradation of these materials. PMID- 16346992 TI - Construction of lactose-assimilating and high-ethanol-producing yeasts by protoplast fusion. AB - The availability of a yeast strain which is capable of fermenting lactose and at the same time is tolerant to high concentrations of ethanol would be useful for the production of ethanol from lactose. Kluyveromyces fragilis is capable of fermenting lactose, but it is not as tolerant as Saccharomyces cerevisiae to high concentrations of ethanol. In this study, we have used the protoplast fusion technique to construct hybrids between auxotrophic strains of S. cerevisiae having high ethanol tolerance and an auxotrophic strain of lactose-fermenting K. fragilis isolated by ethyl methanesulfonate mutagenesis. The fusants obtained were prototrophic and capable of assimilating lactose and producing ethanol in excess of 13% (vol/vol). The complementation frequency of fusion was about 0.7%. Formation of fusants was confirmed by the increased amount of chromosomal DNA per cell. Fusants contained 8 x 10 to 16 x 10 mug of DNA per cell as compared with about 4 x 10 mug of DNA per cell for the parental strains, suggesting that multiple fusions had taken place. PMID- 16346993 TI - Stimulation of tentoxin synthesis by aged-culture filtrates and continued synthesis in the presence of protein inhibitors. AB - Tentoxin, a cyclic tetrapeptide produced by Alternaria alternata (Fries) Keissler, induces chlorosis in certain seedling plants. It can be extracted from culture filtrates of the fungus. Tentoxin production is stimulated and increased by using a mixture of aged culture filtrates and modified Richards solution. Aged culture filtrates can be obtained from 3-week-old or older cultures of A. alternata in modified Richards solution or Pratts solution. A mixture of aged culture filtrate and fresh medium in the ratio 2:3 gives the maximal enhancement of tentoxin production. This growth system provided us with a model for studying the effects of protein synthesis inhibitors on tentoxin production. Two antibiotics which inhibit protein synthesis at the ribosomal level were tested on growth, protein synthesis, and tentoxin production in A. alternata cultures. Cycloheximide at concentrations of 500 mug/ml or emetine at concentrations of 250 mug/ml did not inhibit tentoxin synthesis, although they stopped mycelial growth and protein synthesis of the fungus at the logarithmic growth stage in the enhancement medium. These results led us to conclude that tentoxin, like certain other bioactive cyclic peptides, is synthesized by a nonribosomal peptide synthesis mechanism. PMID- 16346994 TI - Characterization of beta-Glucosidase Activity in Intertidal Marine Sediments. AB - Glycoside derivatives of 4-methylumbelliferone (MUF) were used to characterize the polysaccharidase enzyme systems present in sediments from an intertidal mud flat. The formation of highly fluorescent MUF on hydrolysis of the various glycosides was determined at low substrate concentrations (<1 muM) and with short incubation periods (>5 min). The hydrolysis of MUF-beta-d-glucose in sediments from depth intervals of 0 to 2 cm was insensitive to the presence of oxygen, dissolved sulfide, and iron; magnesium and calcium were stimulatory, however. A pronounced temperature optimum was observed at 40 degrees C, a salinity optimum at 30 per thousand, and a pH optimum at 8.5. Rates of hydrolysis were completely inhibited by the addition of mercuric chloride and sodium azide, but only partially inhibited by toluene and the specific beta-glucosidase inhibitor delta 1,5-gluconolactone. The response to delta-1,5-gluconolactone suggested that about 50% of the observed hydrolysis of MUF-beta-d-glucoside was due to exo- and endoglucanases. A wide variety of hydrolytic activities was observed, with at least some nonspecificity occurring in the case of MUF-beta-d-fucoside. Depth profiles indicated maximal activity in surface sediments with a rapid decline below 2 cm. MUF-glycosides provided a convenlent tool for initial analyses of the dynamics and controls of polymer hydrolysis in marine sediments. PMID- 16346995 TI - Presumed Sexual Isolation in Yeast Populations during Production of Sherrylike Wine. AB - The three stages in wine production in the Montilla-Moriles region of Spain are (i) fermentation, (ii) film formation in vessels, and (iii) aging, all of which selected three yeast populations. All isolates from the three stages were classified as Saccharomyces cerevisiae. The distribution of characteristics of sugar fermentation (sucrose, galactose, maltose), Li tolerance, and Cu tolerance was different in the strains isolated in the three stages. This finding suggests that sexual isolation in the yeast populations prevents the random distribution of taxonomic characters. PMID- 16346996 TI - Adaptation to Hydrogen Sulfide of Oxygenic and Anoxygenic Photosynthesis among Cyanobacteria. AB - Four different types of adaptation to sulfide among cyanobacteria are described based on the differential toxicity to sulfide of photosystems I and II and the capacity for the induction of anoxygenic photosynthesis. Most cyanobacteria are highly sensitive to sulfide toxicity, and brief exposures to low concentrations cause complete and irreversible cessation of CO(2) photoassimilation. Resistance of photosystem II to sulfide toxicity, allowing for oxygenic photosynthesis under sulfide, is found in cyanobacteria exposed to low H(2)S concentrations in various hot springs. When H(2)S levels exceed 200 muM another type of adaptation involving partial induction of anoxygenic photosynthesis, operating in concert with partially inhibited oxygenic photosynthesis, is found in cyanobacterial strains isolated from both hot springs and hypersaline cyanobacterial mats. The fourth type of adaptation to sulfide is found at H(2)S concentrations higher than 1 mM and involves a complete replacement of oxygenic photosynthesis by an effective sulfide-dependent, photosystem II-independent anoxygenic photosynthesis. The ecophysiology of the various sulfide-adapted cyanobacteria may point to their uniqueness within the division of cyanobacteria. PMID- 16346997 TI - Transition from Anoxygenic to Oxygenic Photosynthesis in a Microcoleus chthonoplastes Cyanobacterial Mat. AB - Benthic cyanobacterial mats with the filamentous Microcoleus chthonoplastes as the dominant phototroph grow in oxic hypersaline environments such as Solar Lake, Sinai. The cyanobacteria are in situ exposed to chemical variations between 200 mumol of sulfide liter at night and 1 atm pO(2) during the day. During experimental H(2)S to O(2) transitions the microbial community was shown to shift from anoxygenic photosynthesis, with H(2)S as the electron donor, to oxygenic photosynthesis. Microcoleus filaments could carry out both types of photosynthesis concurrently. Anoxygenic photosynthesis dominated at high sulfide levels, 500 mumol liter, while the oxygenic reaction became dominant when the sulfide level was reduced below 100 to 300 mumol liter (25 to 75 mumol of H(2)S liter). An increasing inhibition of the oxygenic photosynthesis was observed upon transition to oxic conditions from increasing sulfide concentrations. Oxygen built up within the Microcoleus layer of the mat even under 5 mmol of sulfide liter (500 mumol of H(2)S liter) in the overlying water. The implications of such a localized O(2) production in a highly reducing environment are discussed in relation to the evolution of oxygenic photosynthesis during the Proterozoic era. PMID- 16346998 TI - Inhibitory Effect of Autoclaving Whey-Based Medium on Propionic Acid Production by Propionibacterium shermanii. AB - Propionic acid production by Propionibacterium shermanii was compared in pasteurized and autoclaved whey-based media. Propionic acid production decreased with increasing whey concentration in autoclaved media but not in pasteurized media. Increasing the yeast extract concentration from 5 to 10 g/liter greatly reduced the inhibitory effect of autoclaving. PMID- 16346999 TI - Glyphosate-degrading microorganisms from industrial activated sludge. AB - A plating medium was developed to isolate N-phosphonomethylglycine (glyphosate) degrading microorganisms, with glyphosate as the sole phosphorus source. Two industrial biosystems treating glyphosate wastes contained elevated microbial counts on the medium. One purified isolate metabolized glyphosate to aminomethylphosphonic acid, mineralizing this accumulating intermediate during log growth. This microorganism has been identified as a Flavobacterium species. PMID- 16347000 TI - Fungal Metabolism of n-Alkylbenzenes. AB - Isolates of Paecilomyces, Verticillium, Beauveria, and Penicillium species were tested for ability to metabolize a variety of n-alkylbenzenes. Minimum side chain lengths were required for metabolism of these substrates. These were C(4) for the Paecilomyces sp., C(8) for the Verticillium sp., and C(9) for the other two isolates. Growth on dodecylbenzene yielded benzoic and phenylacetic acids as transient intermediates, and these acids supported growth of the isolates. PMID- 16347001 TI - Isolation of Non-O1 Vibrio cholerae Serovars from Oregon Coastal Environments. AB - Water, sediment, and shellfish from three Oregon estuaries were cultured for pathogenic Vibrio species. Non-O1 serovars of V. cholerae were the most common pathogenic Vibrio species recovered. Non-O1 V. cholerae were isolated from all three estuaries sampled, covering an area of about 170 miles along the Oregon coast. Non-O1 V. cholerae were isolated from water and sediment, but not shellfish, at temperatures ranging from 11 to 19 degrees C and salinities of 2.3 to 26 per thousand. Sixteen isolates representing 12 different non-O1 serovars were identified, while four non-O1 V. cholerae isolates failed to react with any of the 54 antisera tested. These results indicate that non-O1 V. cholerae serovars can be found over a large geographic area and under a variety of environmental conditions. These organisms are apparently an autochthonous component of these estuarine microbial communities. PMID- 16347003 TI - Elimination of Viruses and Indicator Bacteria at Each Step of Treatment during Preparation of Drinking Water at Seven Water Treatment Plants. AB - [This corrects the article on p. 1418 in vol. 49.]. PMID- 16347002 TI - Stimulation of Agrobacterium tumefaciens Growth by Azotobacter vinelandii Ferrisiderophores. AB - Azotobacter vinelandii stimulated the growth of Agrobacterium tumefaciens H2, H23, H24, H27, and ATCC 15955 on media containing insoluble iron sources. The Azotobacter vinelandii siderophores appeared to promote Agrobacterium tumefaciens growth by solubilizing mineral iron, and the ferrisiderophores so formed then acted as iron sources for Agrobacterium tumefaciens. Agrobactin, the Agrobacterium siderophore, appeared to be inefficient in solubilizing mineral iron directly. PMID- 16347004 TI - Rhizobins, a group of peptides in the free-amino-Acid pool of the soybean Rhizobium system. AB - Free-living Rhizobium (according to Bergey's Manual of Systematic Bacteriology, [1984, The Williams & Wilkins Co., Baltimore], Bradyrhizobium) japonicum was found to release a peptide into the nutrient media. Soybean nodules contained this peptide and exuded it into the soil. The name "rhizobin A" is suggested for this peptide. Nodules also contained another peptide, rhizobin B, as well as an unidentified, ninhydrin-positive compound, rhizobin C. The three peptides were confined to the free-amino-acid pool of the soluble fraction and eluted consecutively from a cation-exchange column. Rhizobin A was isolated in a highly purified form; its molecular mass was approximately 1,600 daltons as determined by Sephadex gel filtration and mass spectrometry. The amino-acid composition could be determined only approximately, because a long time was necessary for acid hydrolysis, possibly due to unusual linkages. The rhizobin concentration in soybean nodules continually increased during 50 days of growth, from 2 to approximately 400 mug/g (fresh weight). When combined nitrogen was added to nodulated soybean and subsequently removed, nitrogenase activity, nodulation, and nodule growth first decreased and then recovered. The relative amount of rhizobin A followed a similar pattern. Rhizobins were not detected in the roots, stems, and leaves of nodulated soybean plants. They were present in Lupinus nodules, but absent in alder nodules. PMID- 16347005 TI - Improved flotation technique for microscopy of in situ soil and sediment microorganisms. AB - An improved flotation method for microscopy of in situ soil and sediment microorganisms was developed. Microbial cells were released into gellike flotation films that were stripped from soil and sediment aggregates as these aggregates were submerged in 0.5% solutions of polyvinylpyrrolidone. The use of polyvinylpyrrolidone solutions instead of water facilitated the release of films from saturated samples such as aquifer sediments as well as from typical surface soils. In situ microbial morphological characteristics could then be surveyed rapidly by light microscopy of films stained with acridine orange. This method effectively determined the ranges of morphological diversity in a variety of sample types. It also detected microcolonies and other spatial relationships among microbial cells. Only a small fraction (3.4 to 10.1%) of the microflora was released into the flotation films, but plating and direct evaluations by microscopy showed that this fraction was representative of the total population. PMID- 16347006 TI - Modulation of affinity of a marine pseudomonad for toluene and benzene by hydrocarbon exposure. AB - Trace (microgram liter) quantities of either toluene or benzene injected into an amino-acid-limited continuous culture of Pseudomonas sp. strain T2 were utilized immediately with affinities of 2.6 and 6.8 liters g of cells h, respectively, and yielded large amounts of organic products, carbon dioxide, and cells. The immediate utilization of hydrocarbons by hydrocarbon-deprived organisms helps to establish the nutritional value of nonpolar substrates in the environment. The observation of small Michaelis constants for toluene transport led to tests of metabolic competition between hydrocarbons; however, competitive inhibition of toluene metabolism was not found for benzene, naphthalene, xylene, dodecane, or amino acids. Benzene and terpenes were inhibitory at milligram liter concentrations. Toluene was metabolized by a strongly inducible system when compared with benzene. The capacity of toluene to effect larger affinity values increased with exposure time and concentration. The kinetics of induction suggested saturation phenomena, resulting in an induction constant, K(ind), of 96 mug of toluene liter. Maximal induction of amino-acid-grown cells required about 80 h, with the affinity reaching 317 liters g of cells h. PMID- 16347007 TI - Restriction Endonuclease and nif Homology Patterns of Bradyrhizobium japonicum USDA 110 Derivatives With and Without Nitrogen Fixation Competence. AB - DNAs from Bradyrhizobium japonicum USDA 110 derivatives that differ in nitrogen fixing ability produced similar electrophoretic patterns with five different restriction enzymes. Our data support the hypothesis of common ancestry for these derivatives. Derivatives I-110 and L1-110 differed as much as 100-fold in acetylene reduction activity when they were tested with several soybean cultivars in both greenhouse and field experiments. While possessing nodulating ability, derivative L1-110 is deficient in symbiotic nitrogen-fixing ability, whereas derivative I-110 is symbiotically competent. Hybridization of nifDK and nifH probes from B. japonicum to Southern blots of restricted DNAs from strain USDA 110 derivatives produced similar patterns. This finding indicates similar structural gene organization for both derivative I-110 and derivative L1-110 and implies that the difference in symbiotic nitrogen fixation is probably not due to structural gene rearrangements. However, our hybridization data do not rule out the possibility of differences in expression of structural nif genes or alterations in the structure or expression of other genes required for symbiotic nitrogen fixation. PMID- 16347008 TI - Isolation and Characterization of Coproporphyrin Produced by Four Subspecies of Bacillus thuringiensis. AB - It was found by using spectrophotometric, spectrofluorometric, and high-pressure liquid chromatography that four subspecies of Bacillus thuringiensis produce coproporphyrin. The porphyrin isomer was identified as coproporphyrin I for B. thuringiensis subsp. kurstaki (HD1). The porphyrin was isolated both from spores and from a variety of spent growth media. The quantity of porphyrin released by each Bacillus subspecies differed. The rank order of porphyrin production follows: B. thuringiensis subsp. kurstaki HD1 > B. thuringiensis subsp. thuringiensis HD27 > B. thuringiensis subsp. thuringiensis HD41 > B. thuringiensis subsp. darmstadiensis HD199. PMID- 16347009 TI - Identification of Bradyrhizobium japonicum Nodule Isolates from Wisconsin Soybean Farms. AB - One-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis was a more discriminating method than serotyping for identifying strains of Bradyrhizobium japonicum. Analysis of 543 nodule isolates from southeastern Wisconsin soybean farms revealed that none of the isolates were formed by any of the inoculant strains supplied by either of two inoculant companies. Twenty-nine indigenous strains and six inoculant strains were identified. Strain 61A76, the most competitive indigenous strain, formed 21% of the nodules. Indigenous strains 3030, 3058, 0336, and 3052 formed 15, 11, 9, and 9% of the nodules, respectively. These predominant strains were not associated with a particular soybean cultivar, soil type, or farm location. PMID- 16347010 TI - Trichothecene Biosynthesis in Fusarium sporotrichioides: Origin of the Oxygen Atoms of T-2 Toxin. AB - Mass spectral analysis of T-2 toxin formed during the growth of Fusarium sporotrichioides (ATCC 24043) in the presence of H(2)O showed incorporation of up to three O atoms per toxin molecule. The carbonyl oxygens of the acetates at C-4 and C-15 and of the isovalerate at C-8 were derived from H(2)O. Toxin formed in the presence of O molecular oxygen incorporated up to six O atoms per toxin molecule. The overall incorporation was 78 and 92% of toxin molecules labeled for H(2)O and O(2) labeled samples, respectively. The oxygens of position 1, the 12,13-epoxide, and the hydroxyl groups at C-3, C-4, C-8, and C-15 were all derived from molecular oxygen. PMID- 16347011 TI - Optimized batch fermentation of cheese whey-supplemented feedlot waste filtrate to produce a nitrogen-rich feed supplement for ruminants. AB - An optimized batch fermentation process for the conversion of cattle feedlot waste filtrate, supplemented with cheese whey, into a nitrogenous feed supplement for ruminants is described. Feedlot waste filtrate supplemented with cheese whey (5 g of whey per 100 ml) was fermented by the indigenous microbial flora in the feedlot waste filtrate. Ammonium hydroxide was added to the fermentation not only to maintain a constant pH but also to produce ammonium salts of organic acids, which have been shown to be valuable as nitrogenous feed supplements for ruminants. The utilization of substrate carbohydrate at pH 7.0 and 43 degrees C was >94% within 8 h, and the crude protein (total N x 6.25) content of the product was 70 to 78% (dry weight basis). About 66 to 69% of the crude protein was in the form of ammonia nitrogen. Lactate and acetate were the predominant acids during the first 6 to 8 h of fermentation, but after 24 h, appreciable levels of propionate and butyrate were also present. The rate of fermentation and the crude protein content of the product were optimal at pH 7.0 and decreased at a lower pH. For example, fermentation did not go to completion even after 24 h at pH 4.5. Fermentation proceeded optimally at 43 degrees C, less so at 37 degrees C, and considerably more slowly at 23 and 50 degrees C. Concentrations of up to 15 g of cheese whey per 100 ml of feedlot waste filtrate were fermented efficiently. Fermentation of feedlot waste filtrate obtained from animals fed low silage-high grain, high silage-low grain, or dairy rations resulted in similar products in terms of total nitrogen and organic acid composition. PMID- 16347012 TI - Cloning of a Serratia marcescens Gene Encoding Chitinase. AB - Serratia marcescens, a chitinase-producing microorganism, was shown to produce five unique chitinolytic proteins with subunit molecular masses of 21, 36, 48, 52, and 57 kilodaltons. A cosmid library of S. marcescens DNA was constructed in the broad-host-range cosmid pLAFR1 and screened in Escherichia coli for clones capable of degrading chitin. A total of four independent clones (22- to 27 kilobase inserts) were isolated, characterized by restriction endonuclease digestion, and shown to share a common 9.5-kilobase EcoR1 fragment apparently encoding the same 57-kilodalton chitinase, the most abundant chitinase produced by S. marcescens. Chitinase expression from these constructs in both E. coli and Pseudomonas fluorescens 701E1 is apparently driven by an S. marcescens promoter. The significantly higher chitinase levels produced in E. coli relative to those in P. fluorescens 701E1 suggest that E. coli may recognize this promoter sequence more efficiently than P. fluorescens. PMID- 16347013 TI - Sodium-Dependent Growth of Azotobacter chroococcum. AB - The majority of Azotobacter chroococcum strains in a collection obtained from Alberta soils were absolutely dependent on Na for growth. Two strains from the American Type Culture Collection also were either Na dependent or were stimulated by Na. Optimal growth required 0.8 mM Na and was limited at 0.2 to 0.35 mM Na. Growth promoted by 0.8 mM Na was inhibited by Rb > K >> NH(4) but was not affected by Li. Growth inhibition by Rb and K was overcome by increasing the Na concentration present in the medium. Excellent growth in media containing limiting Na was obtained when 25 mM Li or 25 mM Mg was added. Li was significantly more effective in replacing Na than was Mg. Na was required for growth on all C sources (glucose, sucrose, melibiose, and mannitol) and N sources (N(2), NH(4), and NO(3)) and was required throughout the pH range of growth. Cells suspended in Na -deficient medium or in distilled water did not appear to lyse or lose viability. PMID- 16347014 TI - Biotransformation of Unsaturated Long-Chain Fatty Acids by Eubacterium lentum. AB - Eubacterium lentum (33 strains) isomerized the 12-cis double bond of C(18) fatty acids with cis double bonds at C-9 and C-12 into an 11-trans double bond before reduction of the 9-cis double bond. The 14-cis double bond of homo-gamma linolenic acid was isomerized by 29 strains into a 13-trans double bond. The same strains isomerized the 14-cis double bond of arachidonic acid into a 13-trans double bond and then isomerized the 8-cis double bond into a 7-trans double bond; the 13-cis double bond of 10-cis, 13-cis-nonadecadienoic acid was isomerized into a 12-trans double bond. None of these isomerization products was further reduced. Studies with resting cells showed optimal isomerization velocity at a linoleic acid concentration of 37.5 muM; higher concentrations were inhibitory. The pH optimum for isomerization was 7.5 to 8.5. The isomerase was inhibited by the sulfhydryl reagents iodoacetamide, bromoacetate, and N-ethylmaleimide and by the chelators EDTA and 1,10-phenanthroline. PMID- 16347015 TI - Growth and Metabolism of Lactic Acid Bacteria during and after Malolactic Fermentation of Wines at Different pH. AB - Commercially produced red wines were adjusted to pH 3.0, 3.2, 3.5, 3.7, or 4.0 and examined during and after malolactic fermentation for growth of lactic acid bacteria and changes in the concentrations of carbohydrates, organic acids, amino acids, and acetaldehyde. With one exception, Leuconostoc oenos conducted the malolactic fermentation in all wines and was the only species to occur in wines at pH below 3.5. Malolactic fermentation by L. oenos was accompanied by degradation of malic, citric, and fumaric acids and production of lactic and acetic acids. The concentrations of arginine, histidine, and acetaldehyde also decreased at this stage, but the behavior of hexose and pentose sugars was complicated by other factors. Pediococcus parvulus conducted the malolactic fermentation in one wine containing 72 mg of total sulfur dioxide per liter. Fumaric and citric acids were not degraded during this malolactic fermentation, but hexose sugars were metabolized. P. parvulus and species of Lactobacillus grew after malolactic fermentation in wines with pH adjusted above 3.5. This growth was accompanied by the utilization of wine sugars and production of lactic and acetic acids. PMID- 16347016 TI - Comparison of hup trait and intrinsic antibiotic resistance for assessing rhizobial competitiveness axenically and in soil. AB - The competitiveness of dual-strain inocula of cowpea rhizobia for nodulation of Vigna unguiculata (L.) Walp. was studied axenically between one slow-growing strain (P132, HP147, 401, or 22A1) and one fast-growing strain (176A26 or 176A28) at logarithmic inoculum ratios ranging from 10 to 10. Nodule infectivity was determined by multiple intrinsic antibiotic resistance, since both fast-growing strains were sensitive. Different hydrogen uptake (Hup) efficiencies of dual strain inocula allowed for the comparison of an indirect rapid method. Infectivity data based on antibiotic resistance and Hup efficiency were fit to linearized fractile plots of log-normal distributions to determine C(AB) (percent infectivity at a 1:1 inoculum density) or I(50) (inoculum ratio at 50% infectivity). The slow growers were always better competitors and had I(50) values which ranged from 7 to 160,000 and C(AB) values which ranged from 62 to 97%. P132 was the best competitor of all those tested. Antibiotic resistance and Hup efficiency methods were in agreement with 401 (Hup) and 176A26 (Hup), but the Hup efficiency method overestimated the I(50) index with 22A1 (Hup) and 176A28 (Hup). The competition of each of the four slow-growing strains with indigenous rhizobia was examined in Cajanus cajan from three tropical soils. Nodule infectivity for all strains ranged from 42 to 96%, and P132 was the best competitor in all the soils. Hup efficiency overestimated infectivity by about 2 fold when Hup inocula (P132 and HP147) were used but underestimated infectivity by more than 100-fold when Hup inocula (401 and 22A1) were used. Although the Hup trait has limited quantitative usage axenically, it is only qualitative in soil competition studies and can only be used with Hup inocula. PMID- 16347017 TI - Biotransformations of chloroguaiacols, chlorocatechols, and chloroveratroles in sediments. AB - The occurrence of trichloro- and tetrachloroguaiacols, -catechols, and veratroles and their transformation was studied in freshwater and brackish water sediments putatively exposed to bleachery discharge. The samples contained both chloroguaiacols and chlorocatechols, of which >90% could not be removed by simple extraction. The bound concentrations varied and ranged from 550 mug kg of organic C for 3,4,5-trichloroguaiacol to 8,250 mug kg of organic C for tetrachlorocatechol. Chlorinated substrates added to the aqueous phase were rapidly bound to the sediment with K(p) values between 1.3 and 2.8 ml kg of organic C for the chloroguaiacols and chloroveratroles and 22 to 36 ml kg of organic C for the chlorocatechols. Sediment samples incubated aerobically brought about O-methylation of 4,5,6-trichloroguaiacol to 3,4,5-trichloroveratrole in a yield of ca. 25%. Under anaerobic conditions, however, de-O-methylation of both the chloroguaiacols and chloroveratroles took place with synthesis of the corresponding chlorocatechols. In separate experiments, the chlorocatechols were not completely stable under anaerobic conditions, but their ultimate fate has not yet been resolved. Sediment which had been autoclaved twice at 121 degrees C for 20 min was unable to bring about any of these transformations; we therefore conclude that they were mediated by biological processes. These results emphasize that, in determining the fate of chloroguaiacols and related compounds discharged into the aquatic environment, the cardinal roles of sorption to the sediment phase and of the oxygen tension must be taken into account. We propose a hypothetical guaiacol cycle to accommodate our observations. PMID- 16347018 TI - Sulfate reduction relative to methane production in high-rate anaerobic digestion: technical aspects. AB - The effect of different substrates and different levels of sulfate and sulfide on methane production relative to sulfate reduction in high-rate anaerobic digestion was evaluated. Reactors could be acclimated so that sulfate up to a concentration of 5 g of sulfate S per liter did not significantly affect methanogenesis. Higher levels gave inhibition because of salt toxicity. Sulfate reduction was optimal at a relatively low level of sulfate, i.e., 0.5 g of sulfate S per liter, but was also not significantly affected by higher levels. Both acetoclastic and hydrogenotrophic methane-producing bacteria adapted to much higher levels of free H(2)S than the values reported in the literature (50% inhibition occurred only at free H(2)S levels of more than 1,000 mg/liter). High levels of free H(2)S affected the sulfate-reducing bacteria only slightly. Formate and acetate supported the sulfate-reducing bacteria very poorly. In the high-rate reactors studied, intensive H(2)S formation occurred only when H(2) gas or an H(2) precursor such as ethanol was supplied. PMID- 16347019 TI - Sulfate reduction relative to methane production in high-rate anaerobic digestion: microbiological aspects. AB - In the high-rate anaerobic reactors studied (ca. 10 g of chemical oxygen demand [COD] removed per liter of reactor per day), the sulfate-reducing bacteria (SRB) were poor competitors of methane-producing bacteria (MPB), scavenging only on the order of 10 to 20% of the total electron flow. The relatively noncompetitive nature of the SRB in this type of reactor is in sharp contrast to the tendency of the SRB to dominate in natural environments and in other types of anaerobic digesters. Various factors such as the feedback inhibition of H(2)S on the SRB, iron limitation, the origin of the SRB inocula, biokinetics, and thermodynamics were investigated. The outcome of the SRB-MPB competition under the reactor conditions studied appeared to be particularly determined by two factors. The SRB, as predicted by the V(max)-K(m) kinetics, competed most effectively at low substrate levels (<0.5 g of COD per liter). The MPB, however, appeared to colonize and adhere much more effectively to the polyurethane carrier matrix present in the reactor, thus compensating for the apparent lower growth rates. Even if the reactor was initially allowed to be predominantly colonized by SRB, the MPB could regain dominance. PMID- 16347020 TI - Ethylene Dibromide Mineralization in Soils under Aerobic Conditions. AB - 1,2-Dibromoethane (EDB), which is a groundwater contaminant in areas where it was once used as a soil fumigant, was shown to be degraded aerobically by microorganisms in two types of surface soils from an EDB-contaminated groundwater discharge area. At initial concentrations of 6 to 8 mug/liter, EDB was degraded in a few days to near or below the detection limit of 0.02 mug/liter. At 15 to 18 mg/liter, degradation was slower. Bromide ion release at the higher concentrations was 1.4 +/- 0.3 and 2.1 +/- 0.2 molar equivalents for the two soils. Experiments with [C]EDB showed that EDB was converted to approximately equal amounts of CO(2) and apparent cellular carbon; only small amounts of added C were not attributable to these products or unreacted EDB. These results are encouraging, because they indicate that groundwater bacteria may hasten the removal of EDB from contaminated aerobic groundwater supplies. This report also provides evidence for soil-mediated chemical transformations of EDB. PMID- 16347021 TI - Nutrient limitation and adaptation of microbial populations to chemical transformations. AB - Using field-collected periphyton and bacterial isolates, we determined adaptation lag periods for microbial transformation of p-cresol. Lag periods were longer for periphyton samples collected from field sites that were low in dissolved inorganic nitrogen and phosphorus. Moreover, lag periods decreased in samples amended with N or P. Our data suggest that adaptation lag periods for microbial transformation of low concentrations of chemicals may correlate with limiting nutrient concentrations, and this correlation may provide a basis for predictive mathematical modeling of lag periods. PMID- 16347022 TI - Influence of cyanobacterial hyperscum on heterotrophic activity of planktonic bacteria in a hypertrophic lake. AB - The response of the planktonic heterotrophic bacterial community to the buildup and breakdown of a semipermanent, crusted, floating cyanobacterial mat, or hyperscum, that covered 1 to 2 ha was studied in a hypertrophic lake (Hartbeespoort Dam, South Africa). The initial response of bacteria in the main basin to the release of dissolved organic carbon (DOC) from the hyperscum 1 km away was an increase in activity per cell from 35 x 10 to 153 x 10 mug of C cell h for total cell counts, while activity per cell for metabolically active cells increased from 19 x 10 to 85 x 10 mug of C cell h. No major population growth occurred at this stage. Later, with the continuous supply of DOC from the hyperscum, total bacterial numbers increased from 6.6 x 10 to 20 x 10 cells ml, while the activity per cell declined. Metabolically active bacteria followed the same trend. Shorter-term DOC increases caused only increases in bacterial activity per cell. The data from Hartbeespoort Dam demonstrate an interesting and little-documented mechanism by which aquatic bacteria respond to increased DOC concentration and which may be universal for aquatic systems. PMID- 16347023 TI - Attached and free-floating bacterioplankton in howe sound, british columbia, a coastal marine fjord-embayment. AB - Factors which influence the attachment of bacterioplankton to particles (including phytoplankton) were investigated by using (i) water samples removed from a coastal temperate fjord over an annual cycle and (ii) unialgal cultures of Prorocentrum minimum, Dunaliella tertiolecta, and Skeletonema costatum. Silt and salinity levels in this fjord seawater did not appear to influence bacterial attachment, but the percent attached bacteria was inversely related to both chlorophyll a concentrations and primary productivities. During periods of high primary productivities the percent attached bacteria was low, whereas during periods of low, increasing, and declining primary productivities the percent attached bacteria was high. A similar pattern of bacterial attachment was observed when the three phytoplankton were grown as batch cultures. The percent attached bacterial numbers increased upon the initiation of algal growth and after these cells stopped growing, but not while the algae were growing. We suggest that a major factor influencing the attachment of bacterioplankton is the physiological condition of their major nutrient source, the phytoplankton; mainly free-living bacteria are associated with growing phytoplankton, whereas a much greater proportion of the bacteria are attached among senescent phytoplankton populations. PMID- 16347024 TI - Mode of Action of Bipyramidal delta-Endotoxin of Bacillus thuringiensis subsp. kurstaki HD-1. AB - The mode of action of the toxic fragment (P-59) derived from bipyramidal-shaped delta-endotoxin of Bacillus thuringiensis subsp. kurstaki HD-1 on the silkworm Bombyx mori was investigated. An enzyme-linked immunosorbent assay showed that there was no translocation of P-59 from the gut lumen to the hemocoel. When membrane vesicles prepared from silkworm midgut were incubated with P-59, normally smooth surface of vesicles became rough, and patch formation was observed on the surface. Vesicles treated with P-59 tended to agglutinate. The vesicle-denaturing activity of a 130,000-dalton subunit protein of bipyramidal toxin was enhanced by treatment with a gut juice protease of the silkworm. P-59 did not cause any uncoupling effect on mitochondria of the silkworm midgut. These results suggest that the attacking site of this toxin is not the mitochondrion but the cell membrane of the susceptible cell. PMID- 16347025 TI - Evaluation of different assay systems for identification of environmental Aeromonas strains. AB - Important biochemical reactions in conventional tests were compared with counterpart reactions in two multiple test systems, API-20E (Analytab Products, Plainview, N.Y.) and Aeromonas hydrophila medium, to evaluate their accuracy for the identification of motile Aeromonas spp. isolated from fish. In a total of 49 Aeromonas spp. isolates and 10 A. hydrophila reference strains, false-negative or -positive reactions were detected in the Voges-Proskauer test, indole production, gelatinase activity, production of gas, fermentation of arabinose, and lysine decarboxylase reaction. A good correlation was found, among the three identification systems, for the fermentation of mannitol and inositol as well as for the arginine dihydrolase and ornithine decarboxylase tests. The failure of A. hydrophila medium in the detection of gas indicates that this medium is not entirely suitable for defining aerogenic or anaerogenic strains. From the results of the present study, we consider that of the identification method and taxonomic scheme to be adopted for environmental Aeromonas spp. must be standardized. PMID- 16347026 TI - Bacteria associated with the ectoperitrophic space in the midgut of the larva of the midge xylotopus par (Diptera: chironomidae). AB - An ectoperitrophic association of bacteria with the midgut of Xylotopus par larvae was investigated by scanning electron microscopy and transmission electron microscopy. The bacteria attached to the epithelium as a well-defined band in the posterior midgut. They were morphotypically uniform and formed short filaments with endosporelike structures. The consistent presence and well-defined location of the bacteria in a region of the insect digestive tract usually void of microbes indicates a highly evolved symbiotic association, the nature of which is unknown at present. PMID- 16347027 TI - Role for Emulsan in Growth of Acinetobacter calcoaceticus RAG-1 on Crude Oil. AB - When Acinetobacter calcoaceticus RAG-1 was grown together with an emulsan deficient mutant on crude oil, only the emulsan-producing RAG-1 was found to grow, regardless of whether the medium was supplemented with emulsan. The results suggested that the cell-associated form of the bioemulsifier is the biologically active species required for growth on crude oil. A revertant of an emulsan deficient strain was isolated which simultaneously regained the ability to produce both cell-associated and cell-free emulsan as well as the ability to grow on crude oil. PMID- 16347028 TI - Comparison of acridine orange, acriflavine, and bisbenzimide stains for enumeration of bacteria in clear and humic waters. AB - In highly humic water, acridine orange precipitated with dissolved humic matter, resulting in such bright background fluorescence that no bacteria could be seen. With bisbenzimide staining, a similar precipitate was nonfluorescent but obscured many cells. An acriflavine staining method proved useful and reproducible both in clear and in humic waters. Fading of fluorescence was not a problem, and stained samples could be stored after preparation. The fluorescence of cells stained with acriflavine was weaker than that with acridine orange, making counting extremely small cells slightly more difficult with the former stain. PMID- 16347030 TI - Inability of Pseudomonas stutzeri Denitrification Mutants with the Phenotype of Pseudomonas aeruginosa to Grow in Nitrous Oxide. AB - [This corrects the article on p. 1301 in vol. 50.]. PMID- 16347029 TI - Effects of Low-Temperature Acclimation and Oxygen Stress on Tocopherol Production in Euglena gracilis Z. AB - [This corrects the article on p. 1406 in vol. 50.]. PMID- 16347031 TI - Involvement of a Plasmid in Production of Ropiness (Mucoidness) in Milk Cultures by Streptococcus cremoris MS. AB - Curing and genetic transfer experiments showed that lactose-fermenting ability (Lac) and the ability to produce mucoidness in milk cultures (Muc) in Streptococcus cremoris MS were coded on plasmids. The Lac phenotype was associated with a 75.8-megadalton plasmid, pSRQ2201. The Muc phenotype was associated with a 18.5-megadalton plasmid, pSRQ2202. The Lac plasmid, pSRQ2201, was first conjugatively transferred from S. cremoris MS to LacS. lactis ML-3/2.2. Later, the Muc plasmid, pSRQ2202, was conjugatively transferred from Lac MucS. cremoris MS04 to Lac nonmucoid S. lactis transconjugant ML-3/2.201. Subsequently, pSRQ2201 and pSRQ2202 were cotransferred from Lac MucS. lactis transconjugant ML 3/2.202 to Lac, nonmucoid, malty S. lactis 4/4.2 and S. lactis subsp. diacetylactis SLA3.25. Transconjugants showing pSRQ2201 were Lac; those containing pSRQ2202 were Muc. With the transfer of pSRQ2202, the transconjugants S. lactis ML-3/2.202 and S. lactis subsp. diacetylactis SLA3.2501 not only acquired the Muc phenotype but also resistance to bacteriophages, which were lytic to the respective parent strains S. lactis ML-3/2.201 and S. lactis subsp. diacetylactis SLA3.25. PMID- 16347032 TI - Organic matter mineralization with reduction of ferric iron in anaerobic sediments. AB - The potential for ferric iron reduction with fermentable substrates, fermentation products, and complex organic matter as electron donors was investigated with sediments from freshwater and brackish water sites in the Potomac River Estuary. In enrichments with glucose and hematite, iron reduction was a minor pathway for electron flow, and fermentation products accumulated. The substitution of amorphous ferric oxyhydroxide for hematite in glucose enrichments increased iron reduction 50-fold because the fermentation products could also be metabolized with concomitant iron reduction. Acetate, hydrogen, propionate, butyrate, ethanol, methanol, and trimethylamine stimulated the reduction of amorphous ferric oxyhydroxide in enrichments inoculated with sediments but not in uninoculated or heat-killed controls. The addition of ferric iron inhibited methane production in sediments. The degree of inhibition of methane production by various forms of ferric iron was related to the effectiveness of these ferric compounds as electron acceptors for the metabolism of acetate. The addition of acetate or hydrogen relieved the inhibition of methane production by ferric iron. The decrease of electron equivalents proceeding to methane in sediments supplemented with amorphous ferric oxyhydroxides was compensated for by a corresponding increase of electron equivalents in ferrous iron. These results indicate that iron reduction can outcompete methanogenic food chains for sediment organic matter. Thus, when amorphous ferric oxyhydroxides are available in anaerobic sediments, the transfer of electrons from organic matter to ferric iron can be a major pathway for organic matter decomposition. PMID- 16347033 TI - Effects of long-term preservation of frankia strains on infectivity, effectivity, and in vitro nitrogenase activity. AB - Frankia strain HFP ArI3 which had been preserved for 27 months by being lyophilized, frozen in glycerol, or stored in complex medium was successfully used as an inoculum after being subcultured for inducing nodulation and nitrogen fixation of Alnus rubra. Glycerol-preserved HFP ArI3 produced significantly lower rates of nitrogenase activity than did lyophilized or complex-medium-preserved inocula. Bacteria that had been preserved by all three methods were successfully induced to fix atmospheric nitrogen by being cultured in nitrogen-free medium. Subculturing of these cells in nitrogen-free medium a second and third time yielded increasing rates of nitrogenase activity. Initial nitrogenase activity was detected on days 5, 4, and 3 during the first, second, and third subcultures after preservation, respectively. Maximum activity was observed on days 11, 10, and 8 during the first, second, and third subcultures, respectively. A description is given of standard culture techniques used in our laboratory for Frankia isolates, and methods used to distribute Frankia cultures by mail are described. PMID- 16347034 TI - Symbiotic Relationship of Bacteroides cellulosolvens and Clostridium saccharolyticum in Cellulose Fermentation. AB - In coculture, Bacteroides cellulosolvens and Clostridium saccharolyticum fermented 33% more cellulose than did B. cellulosolvens alone. Also, cellulose digestion continued at a maximum rate 48 h longer in coculture. B. cellulosolvens hydrolyzes cellulose and supplies C. saccharolyticum with sugars and a growth factor replaceable by yeast extract. Alone, B. cellulosolvens exhibited an early cessation of growth which was not due to nutrient depletion, low pH, or toxic accumulation of acetic acid, ethanol, lactic acid, H(2), CO(2), cellobiose, glucose, or xylose. However, a 1-h incubation of B. cellulosolvens spent-culture medium with C. saacharolyticum cells starved for growth factor allowed a resumption of B. cellulosolvens growth. The symbiotic relationship of this naturally occurring coculture is one of mutualism, in which the cellulolytic microbe supplies the saccharolytic microbe with nutrients, and in turn the saccharolytic microbe removes a secondary metabolite toxic to the primary microbe. PMID- 16347035 TI - Improved Method of Typing Bradyrhizobium japonicum in Soybean Nodules. AB - An improved method for antibiotic resistance recovery of Bradyrhizobium japonicum from soybean (Glycine max (L.) Merr.) nodules that is simple, time saving, and economical was developed. This technique involves the use of two 96-well microtiter plates as a multinodule sterilization chamber and a template and a third plate as a 16-point replicator constructed with steel nails affixed to the plate with epoxy cement. With this system a team of four technicians could type 3,000 nodules per day. This method was useful in assessing strain establishment and interstrain competition when one or more uniquely labeled strains of B. japonicum were inoculated onto either growth-room- or field-grown soybeans. Contamination was low and reproducibility across replicates approached the theoretical upper limit. Simplicity in design and use made this recovery method especially adaptable for field studies in which large numbers of nodules were required to provide a representative statistical sample offering good precision. PMID- 16347036 TI - Influence of Oxygen and pH on Methanethiol Production from l-Methionine by Brevibacterium linens CNRZ 918. AB - The effects of dissolved oxygen concentration and pH on the growth of Brevibacterium linens CNRZ 918 and its production of methanethiol from l methionine were investigated. Optimal specific methanethiol production was obtained at 25% saturation of dissolved oxygen and at a pH between 8 and 9, whereas optimal cell growth occurred at 50% oxygen saturation and when the pH was maintained constantly at 7. Methanethiol production by nonproliferating bacteria required the presence of l-methionine (7 mM) in the culture medium. This was probably due to the induction of enzyme systems involved in the process. The intracellular concentration of l-methionine seemed to play a key role in this process. B. linens CNRZ 918 tolerated alkaline pHs with a maximal growth pH of approximately 9. Its orange pigmentation seemed to depend on the presence of l methionine in the culture medium and on the concentration of dissolved oxygen. PMID- 16347037 TI - Biochemical function and ecological significance of novel bacterial lipids in deep-sea procaryotes. AB - The fatty acid composition of the membrane lipids in 11 deep-sea bacterial isolates was determined. The fatty acids observed were typical of marine vibrios except for the presence of large amounts of long-chain polyunsaturated fatty acids (PUFAs). These long-chain PUFAs were previously thought to be absent in procaryotes, with the notable exception of a single marine Flexibacter sp. In three barophilic strains tested at 2 degrees C, there was a general increase in the relative amount of PUFAs as pressure was increased from a low growth pressure towards the optimal growth pressure. In Vibrio marinus MP-1, a psychrophilic strain, PUFAs were found to increase as a function of decreasing temperature at constant atmospheric pressure. These results suggest the involvement of PUFAs in the maintenance of optimal membrane fluidity and function over environmentally relevant temperatures and pressures. Furthermore, since these lipids are essential nutrients for higher taxa and are found in large amounts in the lipids of deep-sea vertebrates and invertebrates, an important, specific role for deep sea bacteria in abyssal food webs is implicated. PMID- 16347038 TI - Production and Properties of Xylan-Degrading Enzymes from Cellulomonas uda. AB - Xylan degradation and production of beta-xylanase and beta-xylosidase activities were studied in cultures of Cellulomonas uda grown on purified xylan from birchwood. beta-Xylanase activity was found to be associated with the cells, although in various degrees. The formation of beta-xylanase activity was induced by xylotriose and repressed by xylose. beta-Xylosidase activity was cell bound. Both constitutive and inducible beta-xylosidase activities were suggested. beta Xylanase and beta-xylosidase activities were inhibited competitively by xylose. beta-Xylanase activity had a pronounced optimum pH of 5.8, whereas the optimum pH of beta-xylosidase activity ranged from 5.4 to 6.1. The major products of xylan degradation by a crude preparation of beta-xylanase activity, in decreasing order of amount, were xylobiose, xylotriose, xylose, and small amounts of xylotetraose. This pattern suggests that beta-xylanase activity secreted by C. uda is of the endosplitting type. Supernatants of cultures grown on cellulose showed not only beta-glucanase but also beta-xylanase activity. The latter could be attributed to an endo-1,4-beta-glucanase activity which had a low beta-xylanase activity. PMID- 16347039 TI - Radioisotopic method for measuring cell division rates of individual species of diatoms from natural populations. AB - Silicon is an essential element for diatom frustule synthesis and is usually taken up only by dividing cells. With Ge, a radioactive analog of Si, the cell cycle marker event of frustule formation was identified for individual species of diatom. The frequency of cells within a population undergoing this division event was estimated, and the cell division rate was calculated. In laboratory cultures, these rates of cell division and those calculated from changes in cell numbers were similar. By dual labeling with Ge(OH)(4) and NaHCO(3), rates of cell division and photosynthesis were coincidently measured for diatoms both in laboratory cultures and when isolated from natural populations in estuarine, offshore, and polar environments. These techniques permit the coupling between photosynthesis and cell division to be examined in situ for individual species of diatom. PMID- 16347040 TI - Method for detection of microorganisms that produce gaseous nitrogen oxides. AB - A method was developed to detect NO- or N(2)O-producing bacteria in solid or liquid medium by their ability to oxidize the redox indicator resazurin from its reduced colorless form to its oxidized pink form. The method was sensitive to as little as 35 nM N(2)O or 0.5 nM NO. Ninety-one percent of the colonies that oxidized resazurin on plates also produced N(2)O in slant cultures. Forty-four percent of the colonies that did not oxidize resazurin did produce N(2)O. This percentage was reduced to 15% when colonies in which the coloration was difficult to discern were picked to slants to determine whether they oxidized the slant. The production of N(2)O preceded the oxidation of resazurin by liquid cultures of Escherichia coli and a sludge isolate. With the denitrifying sewage isolate, the disappearance of N(2)O was followed by the return of resazurin to its reduced state. Wolinella succinogenes was found to produce small amounts of N(2)O from NO(3), which resulted in a transient oxidation of resazurin. PMID- 16347041 TI - Transport and metabolism of lactose, glucose, and galactose in homofermentative lactobacilli. AB - A number of species of lactobacilli were examined for their ability to ferment both the glucose and galactose moieties of lactose. Lactobacillus helveticus strains metabolized both the glucose and galactose moieties, whereas L. bulgaricus, L. lactis, and L. acidophilus strains metabolized only the glucose moiety and released galactose into the growth medium. All four species tested contained beta-galactosidase activity, and no significant phospho-beta galactosidase activity was observed. L. bulgaricus and L. helveticus had a phosphoenolpyruvate (PEP):glucose phosphotransferase system for the uptake of glucose, but no evidence for a PEP:lactose phosphotransferase or PEP:galactose phosphotransferase system was obtained. PMID- 16347042 TI - Nitrogen Fixation Associated with Development and Localization of Mixed Populations of Cellulomonas sp. and Azospirillum brasilense Grown on Cellulose or Wheat Straw. AB - Mixed cultures of Cellulomonas sp. and Azospirillum brasilense were grown with straw or cellulose as the carbon source under conditions favoring the fixation of atmospheric nitrogen. Rapid increases in cell numbers, up to 10 cells per g of substrate, were evident after 4 and 5 days of incubation at 30 degrees C for cellulose and straw, respectively. Nitrogen fixation (detected by acetylene reduction measured on parallel cultures) commenced after 2 and 4 days of incubation for straw and cellulose, respectively, and continued for the duration of the experiment. Pure cultures of Cellulomonas sp. showed an increase in cell numbers, but CO(2) production was low, and acetylene reduction was not detected on either cellulose or straw. Pure cultures of A. brasilense on cellulose showed an initial increase in cell numbers (10 cells per g of substrate) over 4 days, followed by a decline presumably caused by the exhaustion of available carbon substrate. On straw, A. brasilense increased to 10 cells per g of substrate over 5 days and then declined slowly; this growth was accompanied by acetylene reduction. Scanning electron micrographs of straw incubated with a mixed culture under the above conditions for 8 days showed cells of both species in close proximity to each other. Evidence was furnished that the close spatial relationship of cells from the two species facilitated the mutually beneficial association between them and thus increased the efficiency with which the products of straw breakdown were used for nitrogen fixation. PMID- 16347043 TI - Comparison of Two Cellulomonas Strains and Their Interaction with Azospirillum brasilense in Degradation of Wheat Straw and Associated Nitrogen Fixation. AB - A mutant strain of Cellulomonas sp. CS1-17 was compared with Cellulomonas gelida 2480 as the cellulolytic component of a mixed culture which was responsible for the breakdown of wheat straw to support asymbiotic nitrogen fixation by Azospirillum brasilense Sp7 (ATCC 29145). Cellulomonas sp. strain CSI-17 was more efficient than was C. gelida in cellulose breakdown at lower oxygen concentrations and, in mixed culture with A. brasilense, it supported higher nitrogenase activity (C(2)H(2) reduction) and nitrogen fixation with straw as the carbon source. Based on gravimetric determinations of straw breakdown and total N determinations, the efficiency of nitrogen fixation was 72 and 63 mg of N per g of straw utilized for the mixtures containing Cellulomonas sp. and C. gelida, respectively. Both Cellulomonas spp. and Azospirillum spp. exhibited a wide range of pH tolerance. When introduced into sterilized soil, the Cellulomonas sp. Azospirillum brasilense association was more effective in nitrogen fixation at a pH of 7.0 than at the native soil pH (5.6). This was also true of the indigenous diazotrophic microflora of this soil. The potential implications of this work to the field situation are discussed. PMID- 16347044 TI - Cloning and Expression in Escherichia coli of the Polysaccharide Depolymerase Associated with Bacteriophage-Infected Erwinia amylovora. AB - The bacteriophage-encoded polysaccharide depolymerase produced in Erwinia amylovora has been cloned and expressed in Escherichia coli. The bacteriophage ERA103 genome was observed to consist of five EcoRI fragments, labeled as follows: A, 7.5 kilobases (kb); B, 5.0 kb; C, 2.7 kb; D, 2.1 kb; and E, 1.8 kb. A restriction map for ERA103 was also prepared. Each of the fragments were cloned into the positive-selection vector pOP203(A(2)) and pBR322. PMID- 16347045 TI - Asymbiotic acetylene reduction by a fast-growing cowpea Rhizobium strain with nitrogenase structural genes located on a symbiotic plasmid. AB - A procedure was designed which enabled the detection of ex planta nitrogenase activity in the fast-growing cowpea Rhizobium strain IHP100. Nitrogenase activity in agar culture under air occurred at a rate similar to that found for Bradyrhizobium strain CB756 but lower than that for Rhizobium strain ORS571. Hybridization studies showed that both nod and nif genes were located on a 410 kilobase Sym plasmid in strain IHP100. PMID- 16347046 TI - Inoculation response of legumes in relation to the number and effectiveness of indigenous Rhizobium populations. AB - The response of legumes to inoculation with rhizobia can be affected by many factors. Little work has been undertaken to examine how indigenous populations or rhizobia affect this response. We conducted a series of inoculation trials in four Hawaiian soils with six legume species (Glycine max, Vigna unguiculata, Phaseolus lunatus, Leucaena leucocephala, Arachis hypogaea, and Phaseolus vulgaris) and characterized the native rhizobial populations for each species in terms of the number and effectiveness of the population for a particular host. Inoculated plants had, on average, 76% of the nodules formed by the inoculum strain, which effectively eliminated competition from native strains as a variable between soils. Rhizobia populations ranged from less than 6 x 10/g of soil to 1 x 10/g of soil. The concentration of nitrogen in shoots of inoculated plants was not higher than that in uninoculated controls when the most probable number MPN counts of rhizobia were at or above 2 x 10/g of soil unless the native population was completely ineffective. Tests of random isolates from nodules of uninoculated plants revealed that within most soil populations there was a wide range of effectiveness for N(2) fixation. All populations had isolates that were ineffective in fixing N(2). The inoculum strains generally did not fix more N(2) than the average isolate from the soil population in single-isolate tests. Even when the inoculum strain proved to be a better symbiont than the soil rhizobia, there was no response to inoculation. Enhanced N(2) fixation after inoculation was related to increased nodule dry weights. Although inoculation generally increased nodule number when there were less than 1 x 10 rhizobia per g of soil, there was no corresponding increase in nodule dry weight when native populations were effective. Most species compensated for reduced nodulation in soils with few rhizobia by increasing the size of nodules and therefore maintaining a nodule dry weight similar to that of inoculated plants with more nodules. Even when competition by native soil strains was overcome with a selected inoculum strain, it was not always possible to enhance N(2) fixation when soil populations were above a threshold number and had some effective strains. PMID- 16347047 TI - Purification and Properties of beta-N-Acetylhexosaminidase from Mucor fragilis Grown in Bovine Blood. AB - Mucor fragilis grown on bovine blood powder as the sole carbon source abundantly produced beta-N-acetylhexosaminidase. The enzyme activity was several times higher than that of a culture obtained with glucose medium. The enzyme had two different molecular weight forms. The high-molecular-weight form had somewhat higher beta-N-acetylgalactosaminidase activity than the lower-molecular-weight enzyme which had beta-N-acetylgalactosaminidase activity equivalent to about 40% of its beta-N-acetylglucosaminidase activity. Bovine blood seemed to induce both enzymes, but N-acetylamino sugars specifically induced the low-molecular-weight form. N-Acetylgalactosamine had an especially marked effect on activity. The low molecular-weight form of enzyme was purified from the culture filtrate by fractionation with ammonium sulfate and various column chromatographies. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis. The optimum pH was 4.0 to 5.0 for beta-N-acetylglucosaminidase activity and 5.5 to 6.5 for beta-N-acetylgalactosaminidase activity. The enzyme hydrolyzed natural substrates such as di-N-acetylchitobiose, tri-N acetylchitotriose, and a glycopeptide obtained by modification of fetuin. PMID- 16347048 TI - Improvement of an l-Leucine-Producing Mutant of Brevibacterium lactofermentum 2256 by Genetically Desensitizing It to alpha-Acetohydroxy Acid Synthetase. AB - Genetic improvement of l-leucine productivity in strain 218, an ile 2 thiazolealanine-resistant mutant of Brevibacterium lactofermentum 2256, was attempted. In strain 218, which produced 28 mg of l-leucine per ml from 13% glucose, alpha-isopropylmalate synthetase was genetically desensitized and derepressed to the effect of l-leucine, whereas alpha-acetohydroxy acid synthetase remained unaltered, although it could be derepressed phenotypically by limiting the isoleucine concentration in the culture. From strain 218 we isolated 103 mutants resistant to beta-hydroxyleucine (4 mg/ml). Among these, three were found to produce mere l-leucine than the parent. The alpha-acetohydroxy acid synthetase of all three mutant strains was found to be genetically desensitized to all of the branched-chain amino acids l-isoleucine, l-valine, and l-leucine. The repression mechanism in alpha-acetohydroxy acid synthetase formation was the same as in the parent strain. The improved strains typically produced 34 mg of l leucine per ml, the highest productivity ever reported. PMID- 16347049 TI - Production and Characteristics of Avicel-Disintegrating Endoglucanase from a Protease-Negative Humicola grisea var. thermoidea Mutant. AB - Mutational experiments were performed to decrease the protease productivity of Humicola grisea var. thermoidea YH-78 using UV light and N-methyl-N'-nitro-N nitrosoguanidine. A protease-negative mutant, no. 140, exhibited higher endoglucanase activity than the parent strain in mold bran culture at 50 degrees C for 4 days. The culture extract rapidly disintegrated filter paper but produced a small amount of reducing sugar. About 30% of total endoglucanase activity in the extract was adsorbed onto Avicel. The electrophoretically homogeneous preparation of Avicel-adsorbable endoglucanase (molecular weight, 128,000) showed intensive filter-paper-disintegrating activity but did not release reducing sugar. The preparation also exhibited a highly synergistic effect with the cellulase preparation from Trichoderma reesei in the hydrolysis of microcrystalline cellulose. This endoglucanase was observed via scanning electron microscopy to disintegrate Avicel fibrils layer by layer from the surface, yielding thin sections with exposed chain ends. A mutant, no. 191, producing higher protease activity and an Avicel-unadsorbable, Avicel-nondisintegrating endoglucanase was isolated. The purified enzyme (molecular weight, 63,000) showed no disintegrating activity on filter paper and Avicel and a less synergistic effect with the T. reesei cellulase in hydrolyzing microcrystalline cellulose than did the former enzyme. Endoglucanase was therefore divided into two types, Avicel disintegrating and Avicel nondisintegrating. PMID- 16347050 TI - Growth of methanogenic bacteria in pure culture with 2-propanol and other alcohols as hydrogen donors. AB - Two types of mesophilic, methanogenic bacteria were isolated in pure culture from anaerobic freshwater and marine mud with 2-propanol as the hydrogen donor. The freshwater strain (SK) was a Methanospirillum species, the marine, salt-requiring strain (CV), which had irregular coccoid cells, resembled Methanogenium sp. Stoichiometric measurements revealed formation of 1 mol of CH(4) by CO(2) reduction, with 4 mol of 2-propanol being converted to acetone. In addition to 2 propanol, the isolates used 2-butanol, H(2), or formate but not methanol or polyols. Acetate did not serve as an energy substrate but was necessary as a carbon source. Strain CV also oxidized ethanol or 1-propanol to acetate or propionate, respectively; growth on the latter alcohols was slower, but final cell densities were about threefold higher than on 2-propanol. Both strains grew well in defined, bicarbonate-buffered, sulfide-reduced media. For cultivation of strain CV, additions of biotin, vitamin B(12), and tungstate were necessary. The newly isolated strains are the first methanogens that were shown to grow in pure culture with alcohols other than methanol. Bioenergetic aspects of secondary and primary alcohol utilization by methanogens are discussed. PMID- 16347051 TI - Aerobic Hydrogen Accumulation by a Nitrogen-Fixing Cyanobacterium, Anabaena sp. AB - Hydrogen evolution by a nitrogen-fixing cyanobacterium, Anabaena sp. strain N 7363, was tested in order to develop a water biophotolysis system under aerobic conditions. A culture of the strain supplemented with carbon dioxide under an air atmosphere evolved hydrogen and oxygen gas, which reached final concentrations of 9.7 and 69.8%, respectively, after 12 days of incubation. Hydrogen uptake activity was not observed during incubation, and nitrogenase was thought to be the sole enzyme responsible for the hydrogen evolution. PMID- 16347052 TI - Enhancement of Wheat Root Colonization and Plant Development by Azospirillum brasilense Cd. Following Temporary Depression of Rhizosphere Microflora. AB - Inoculation of wheat with Azospirillum brasilense, combined with the application of four fungal and bacterium-inhibiting substances to which A. brasilense is resistant in the soil, decreased the rhizosphere population, while it increased wheat root colonization by A. brasilense, even in cases of poor inoculation. The inoculation significantly increased the following wheat plant parameters as well: plant dry weight, number of tillers per plant, spikelet fertility, harvest index, and grain yield. This model may provide a new approach to improve control of root colonization by beneficial bacteria. PMID- 16347053 TI - Fermentation of aqueous plant seed extracts by lactic Acid bacteria. AB - The effects of lactic acid bacterial fermentation on chemical and physical changes in aqueous extracts of cowpea (Vigna unguiculata), peanut (Arachis hypogea), soybean (Glycine max), and sorghum (Sorghum vulgare) were studied. The bacteria investigated were Lactobacillus helveticus, L. delbrueckii, L. casei, L. bulgaricus, L. acidophilus, and Streptococcus thermophilus. Organisms were inoculated individually into all of the seed extracts; L. bulgaricus and S. thermophilus were also evaluated together as inocula for fermenting the legume extracts. During fermentation, bacterial population and changes in titratable acidity, pH, viscosity, and color were measured over a 72-h period at 37 degrees C. Maximum bacterial populations, titratable acidity, pH, and viscosity varied depending upon the type of extract and bacterial strain. The maximum population of each organism was influenced by fermentable carbohydrates, which, in turn, influenced acid production and change in pH. Change in viscosity was correlated with the amount of protein and titratable acidity of products. Color was affected by pasteurization treatment and fermentation as well as the source of extract. In the extracts inoculated simultaneously with L. bulgaricus and S. thermophilus, a synergistic effect resulted in increased bacterial populations, titratable acidity, and viscosity, and decreased pH in all the legume extracts when compared to the extracts fermented with either of these organisms individually. Fermented extracts offer potential as substitutes for cultured dairy products. PMID- 16347054 TI - Influence of Location, Host Cultivar, and Inoculation on the Composition of Naturalized Populations of Rhizobium meliloti in Medicago sativa Nodules. AB - A phage typing system was used to evaluate the composition of indigenous populations of Rhizobium meliloti inhabiting nodules of Medicago sativa cultivars grown with and without inoculation at two field sites during 1983 and 1984. Soil at both locations contained established populations of R. meliloti at planting. Analysis of 1,920 nodule isolates revealed 55 unique phage types of indigenous R. meliloti at one site and 65 indigenous types at the other location. The distributions of phage types differed markedly between locations. At one site, the nodule population was dominated by two phage types; seven others occurred consistently but at lower frequency, and the remainder were encountered infrequently. No indigenous types predominated at the other location, although nine occurred more frequently than the remaining types. Indigenous R. meliloti predominated in nodules from inoculated plots at both sites, with inoculant recovery varying between 10 and 38% in each of two years. The frequency of occurrence of particular phage types at one location was significantly influenced by both M. sativa cultivar and inoculation. At this location, the interaction of cultivar and inoculation on the incidence of phage types suggests that the presence of an inoculant strain differentially affected nodule occupancy of M. sativa cultivars by members of the indigenous R. meliloti population. At both sites, the frequency of specific phage types differed between years. The data emphasize the importance of understanding the ecology and characteristics of indigenous Rhizobium populations as a prerequisite for elucidating problems of inoculant establishment and persistence in competitive situations. PMID- 16347055 TI - Alginate beads as synthetic inoculant carriers for slow release of bacteria that affect plant growth. AB - Uniform synthetic beads were developed as carriers for the bacterial inoculation of plants. The beads are made of sodium alginate and skim milk and contain a large reservoir of bacterial culture which releases the bacteria at a slow and constant rate. The beads are biodegradable and produce no environmental pollution. The strength of the beads, the rate of bacterial release, and the time of their survival in the soil can be controlled by several hardening treatments. The final product, lyophilized beads, is simple to use and is applied to the seeds concomitantly with sowing. The released bacteria are available for root colonization immediately at seed germination. Dry beads containing bacteria can be stored at ambient temperature over a long period without loss of bacterial content; storage requires a limited space, and the quality control of a number of bacteria in the bead is simple. The level of plant inoculation with beads was similar to that with previously used peat inoculants, but the former method yielded more consistent results, as the frequency of inoculated plants was much higher. The former method provides a different approach for inoculation of plants with beneficial rhizosphere bacteria. PMID- 16347056 TI - Bioaccumulation of Germanium by Pseudomonas putida in the Presence of Two Selected Substrates. AB - The uptake of germanium by Pseudomonas putida ATCC 33015 was studied in the presence of catechol or acetate or both as representative substrates differing in their ability to form complexes with this element. The bacteria were taken from a batch culture grown on acetate as the sole carbon source. Cells introduced into a medium containing germanium and either catechol or a mixture of catechol and acetate accumulated germanium in a biphasic way. After a lower level of accumulation that corresponded to the value obtained in the presence of acetate was reached, a further increase in the germanium content up to a higher saturation level was observed. The appearance of the second step of accumulation, which corresponded to the linear degradation of catechol, proved that catechol facilitated the transport of germanium into the cells through the nonspecific uptake of the germanium-catechol complex by an inducible catechol transport system. PMID- 16347057 TI - Methodology for estimating numbers of free-living and attached bacteria in estuarine water. AB - A fundamental problem in estuarine microbiology studies is the accurate determination of the density in the water column of both free-living bacteria and those attached to suspended particulate matter. When a water sample is filtered and the filter is viewed by epifluorescence microscopy, counts can be made of the numbers of bacteria which are seen on the filter background (free-living) and those which appear to lie on sediment particles (both free-living and attached). With only the additional knowledge of the proportion of the filter area covered by particles (a quantity that is straightforwardly determined by stereological point counting), results from geometric probability were used to determine the expected number of bacteria which are hidden by particles and hence to provide an estimation scheme for the true densities of free-living and attached bacteria. Variance equations based on a Taylor series are given, and a partial check of the method is attempted with controlled mixtures of bacteria and sediment. An alternative procedure is also proposed, in which the natural attached/free-living ratio is altered by an intervention experiment, allowing an estimation which is less model dependent but more labor intensive. Both methods are applied to a series of samples from the Tamar estuary, United Kingdom, taken in April 1985. A notable conclusion is that there are always more free-living than attached bacteria in the water column throughout the estuary. PMID- 16347058 TI - Phosphotransferase Activity in Clostridium acetobutylicum from Acidogenic and Solventogenic Phases of Growth. AB - Clostridium acetobutylicum cells, when energized with fructose, transported and phosphorylated the glucose analog 2-deoxyglucose by a phosphoenolpyruvate dependent phosphotransferase (PT) system. Butanol up to 2% did not inhibit PT activity, although its chaotropic effect on the cell membrane caused cellular phosphoenolpyruvate and the 2-deoxyglucose-6-phosphate to leak out. Cells harvested from the solventogenic phase of batch growth had a significantly lower PT activity than did cells from the acidogenic phase. PMID- 16347059 TI - Improved agar bottle plate for isolation of methanogens or other anaerobes in a defined gas atmosphere. AB - A bottle plate for the cultivation of methanogens or other organisms in a defined pressurized-gas atmosphere was developed. The bottle provides the convenience of an agar streak plate, solves the problem of the water exudate from agar medium, and provides a convenient way of adding or sampling a defined gas atmosphere. PMID- 16347060 TI - Adsorption of Agrobacterium tumefaciens to Susceptible Potato Tissues: a Physisorption Process. AB - Adsorption of Agrobacterium tumefaciens cells to potato tuber disks reached equilibrium after coincubation for about 30 min. More than 10% of the number of bacteria bound at equilibrium were adsorbed within 30 s. Adsorption isotherms obtained at three temperatures showed that the heat of adsorption was nearly zero. PMID- 16347061 TI - Catabolism of Amino Acids by Megasphaera elsdenii LC1. AB - The amino acids in an acid hydrolysate of casein were catabolized more extensively by Megasphaera elsdenii than those in an enzymic hydrolysate. Threonine and serine were most actively degraded, but no resultant increase in growth yield occurred. Branched-chain volatile fatty acid production, which increased as the dilution rate of a glucose-limited chemostat decreased, seemed to be associated with maintenance rather than with growth. PMID- 16347062 TI - Binding of Germanium to Pseudomonas putida Cells. AB - The binding of germanium to Pseudomonas putida ATCC 33015 was investigated by using whole intact cells grown in a medium supplemented with GeO(2) and catechol or acetate. Electron-microscopic examination of the control and metal-loaded samples revealed that germanium was bound within the cell envelope. A certain number of small electron-dense deposits of the bound element were found in the cytoplasm when the cells were grown in the presence of GeO(2) and catechol. The study of germanium distribution in cellular fractions revealed that catechol facilitated the intracellular accumulation of this element. PMID- 16347064 TI - Enumeration by DNA Colony Hybridization of Virulent Yersinia enterocolitica Colonies in Artificially Contaminated Food. AB - [This corrects the article on p. 441 in vol. 51.]. PMID- 16347063 TI - Analysis of Rhizobium meliloti Sym Mutants Obtained by Heat Treatment. AB - Deletions in the pSym megaplasmid of Rhizobium meliloti were produced at a high frequency, and their lengths varied according to incubation temperature. Morphological differentiation into large and small colonies occurred after heat treatment. Small colonies elicited pseudonodules on alfalfa roots. PMID- 16347065 TI - Bioreactor for the study of defined interactions of toxic metals and biofilms. AB - [This corrects the article on p. 1155 in vol. 50.]. PMID- 16347066 TI - Symbiotic Effectiveness and Host-Strain Interactions of Rhizobium fredii USDA 191 on Different Soybean Cultivars. AB - Nodulation, acetylene reduction activity, dry matter accumulation, and total nitrogen accumulation by nodulated plants growing in a nitrogen-free culture system were used to compare the symbiotic effectiveness of the fast-growing Rhizobium fredii USDA 191 with that of the slow-growing Bradyrhizobium japonicum USDA 110 in symbiosis with five soybean (Glycine max (L.) Merr.) cultivars. Measurement of the amount of nitrogen accumulated during a 20-day period of vegetative growth (28 to 48 days after transplanting) showed that USDA 110 fixed 3.7, 39.1, 4.6, and 57.3 times more N(2) than did USDA 191 with cultivars Pickett 71, Harosoy 63, Lee, and Ransom as host plants, respectively. With the unimproved Peking cultivar as the host plant, USDA 191 fixed 3.3 times more N(2) than did the USDA 110 during the 20-day period. The superior N(2) fixation capability of USDA 110 with the four North American cultivars as hosts resulted primarily from higher nitrogenase activity per unit nodule mass (specific acetylene reduction activity) and higher nodule mass per plant. The higher N(2)-fixation capability of USDA 191 with the Peking cultivar as host resulted primarily from higher nodule mass per plant, which was associated with higher nodule numbers. There was significant variation in the N(2)-fixation capabilities of the four North American cultivar-USDA 191 symbioses. Pickett 71 and Lee cultivars fixed significantly more N(2) in symbiosis with USDA 191 than did the Harosoy 63 and Ransom cultivars. This quantitative variation in N(2)-fixation capability suggests that the total incompatibility (effectiveness of nodulation and efficiency of N(2) fixation) of host soybean plants and R. fredii strains is regulated by more than one host plant gene. These results indicate that it would not be prudent to introduce R. fredii strains into North American agricultural systems until more efficient N(2)-fixing symbioses between North American cultivars and these fast-growing strains can be developed. When inoculum containing equal numbers of USDA 191 and of strain USDA 110 was applied to the unimproved Peking cultivar in Perlite pot culture, 85% of the 160 nodules tested were occupied by USDA 191. With Lee and Ransom cultivars, 99 and 85% of 140 and 96 nodules tested, respectively, were occupied by USDA 110. PMID- 16347067 TI - Variables affecting two electron transport system assays. AB - Several methodological variables were critical in two commonly used electron transport activity assays. The dehydrogenase assay based on triphenyl formazan production exhibited a nonlinear relationship between formazan production (dehydrogenase activity) and sediment dilution, and linear formazan production occurred for 1 h in sediment slurries. Activity decreased with increased time of sediment storage at 4 degrees C. Extraction efficiencies of formazan from sediment varied with alcohol type; methanol was unsatisfactory. Phosphate buffer (0.06 M) produced higher activity than did either U.S. Environmental Protection Agency reconstituted hard water or Tris buffer sediment diluents. Intracellular formazan crystals were dissolved within minutes when in contact with immersion oil. Greater crystal production (respiration) detected by a tetrazolium salt assay occurred at increased substrate concentrations. Test diluents containing macrophyte exudates produced greater activity than did phosphate buffer, U.S. Environmental Protection Agency water, or ultrapure water diluents. Both assays showed decreases in sediment or bacterial activity through time. PMID- 16347068 TI - Relative rates of nitric oxide and nitrous oxide production by nitrifiers, denitrifiers, and nitrate respirers. AB - Biogenic emissions of nitric and nitrous oxides have important impacts on the photochemistry and chemistry of the atmosphere. Although biogenic production appears to be the overwhelming source of N(2)O, the magnitude of the biogenic emission of NO is very uncertain. In soils, possible sources of NO and N(2)O include nitrification by autotrophic and heterotrophic nitrifiers, denitrification by nitrifiers and denitrifiers, nitrate respiration by fermenters, and chemodenitrification. The availability of oxygen determines to a large extent the relative activities of these various groups of organisms. To better understand this influence, we investigated the effect of the partial pressure of oxygen (pO(2)) on the production of NO and N(2)O by a wide variety of common soil nitrifying, denitrifying, and nitrate-respiring bacteria under laboratory conditions. The production of NO per cell was highest by autotrophic nitrifiers and was independent of pO(2) in the range tested (0.5 to 10%), whereas N(2)O production was inversely proportional to pO(2). Nitrous oxide production was highest in the denitrifier Pseudomonas fluorescens, but only under anaerobic conditions. The molar ratio of NO/N(2)O produced was usually greater than unity for nitrifiers and much less than unity for denitrifiers. Chemodenitrification was the major source of both the NO and N(2)O produced by the nitrate respirer Serratia marcescens. Chemodenitrification was also a possible source of NO and N(2)O in nitrifier cultures but only when high concentrations of nitrite had accumulated or were added to the medium. Although most of the denitrifiers produced NO and N(2)O only under anaerobic conditions, chemostat cultures of Alcaligenes faecalis continued to emit these gases even when the cultures were sparged with air. Based upon these results, we predict that aerobic soils are primary sources of NO and that N(2)O is produced only when there is sufficient soil moisture to provide the anaerobic microsites necessary for denitrification by either denitrifiers or nitrifiers. PMID- 16347069 TI - Microbial 7alpha-hydroxylation of 3-ketobisnorcholenol. AB - The transformation of 22-hydroxy-23,24-bisnorchol-4-en-3-one to 7alpha-22 dihydroxy-23,24-bisnorchol-4-en-3-one by Botryodiploida theobromae, Lasiodiplodia theobromae, and various Botryosphaeria strains is described. Factors affecting the reaction were incubation temperature, sonication of the substrate, and addition of 2,2'-dipyridyl, extra carbohydrate, and Amberlite XAD-7. The enzyme responsible for the reaction appeared to be very specific and was not characteristic of all members of the genera listed above. PMID- 16347070 TI - Degradation of Pyridine by Micrococcus luteus Isolated from Soil. AB - An organism capable of growth on pyridine was isolated from soil by enrichment culture techniques and identified as Micrococcus luteus. The organism oxidized pyridine for energy and released N contained in the pyridine ring as ammonium. The organism could not grow on mono- or disubstituted pyridinecarboxylic acids or hydroxy-, chloro-, amino-, or methylpyridines. Cell extracts of M. luteus could not degrade pyridine, 2-, 3-, or 4-hydroxypyridines or 2,3-dihydroxypyridine, regardless of added cofactors or cell particulate fraction. The organism had a NAD-linked succinate-semialdehyde dehydrogenase which was induced by pyridine. Cell extracts of M. luteus had constitutive amidase activity, and washed cells degraded formate and formamide without a lag. These data are consistent with a previously reported pathway for pyridine metabolism by species of Bacillus, Brevibacterium, and Corynebacterium. Cells of M. luteus were permeable to pyridinecarboxylic acids, monohydroxypyridines, 2,3-dihydroxypyridine, and monoamino- and methylpyridines. The results provide new evidence that the metabolism of pyridine by microorganisms does not require initial hydroxylation of the ring and that permeability barriers do not account for the extremely limited range of substrate isomers used by pyridine degraders. PMID- 16347071 TI - Population Dynamics of Chlamydomonas sajao and Its Influence on Soil Aggregate Stabilization in the Field. AB - Chlamydomonas sajao, a single-celled, eucaryotic microalga, was inoculated onto replicated field plots cropped to corn at two rates (5 x 10 and 5 x 10 log-phase cells ha) to assess colonization, reproduction, and persistence, changes in soil carbohydrate content, and wet stability of 0.92- to 1.68-mm-sized aggregates from the surface 2- to 3-mm soil veneer. The most-probable-number technique and extraction and fluorometric quantification of chlorophyll were used as indices of microalgal abundance. Cell numbers ranged up to 6.9 x 10 g of soil by most probable number and were significantly greater on high-rate than on low-rate and control plots over a 10-week period. Chlorophyll content and most probable number were positively correlated (r = 0.64) for the high-rate plots but not for low rate plots or the controls. Increased wet aggregate stability measurements (33 to 77%) for the high-rate plots during the growing season were significantly greater than for low-rate and control plots, which were not different from each other. The work leads support to the hypothesis that mass-cultured palmelloid microalgae are a feasible means for conditioning soil biologically. PMID- 16347072 TI - Common components of industrial metal-working fluids as sources of carbon for bacterial growth. AB - Water-based metal-working fluids used in large-scale industrial operations consist of many components, but in the most commonly used formulations only three classes of components are present in high enough concentrations that they could, in principle, provide enough carbon to support the high bacterial densities (10 CFU/ml) often observed in contaminated factory fluids. These components are petroleum oil (1 to 5%), petroleum sulfonates (0.1 to 0.5%), and fatty acids (less than 0.1%, mainly linoleic and oleic acids supplied as tall oils). We isolated pure strains of predominating bacteria from contaminated reservoirs of two metal-working systems and randomly selected 12 strains which we tested in liquid culture for growth with each of the metal-working fluid components as the sole source of carbon. Of the 12 strains, 7 reached high density (10 CFU/ml from an initial inoculum of less than 2 x 10) in 24 h, and 1 strain did the same in 48 h with 0.05% oleic or linoleic acid as the carbon source. These same strains also grew on 1% naphthenic petroleum oil but required up to 72 h to reach densities near 10 CFU/ml. One strain grew slightly and the others not at all on the petroleum sulfonates. The four remaining strains did not grow on any of the components, even though they were among the predominating bacteria in the contaminated system. Of the seven strains that grew best on the fatty acids and on the naphthenic petroleum oil, five were tentatively identified as Acinetobacter species and two were identified as Pseudomonas species. Four of the bacteria that did not grow were tentatively identified as species of Pseudomonas, and one could not be identified. PMID- 16347073 TI - Characterization of Leucine Auxotrophs of the White Rot Basidiomycete Phanerochaete chrysosporium. AB - Six leucine auxotrophic strains of the white rot basidiomycete Phanerochaete chrysosporium were characterized genetically and biochemically. Complementation studies involving the use of heterokaryons identified three leucine complementation groups. Since all of the leucine auxotrophs grew on minimal medium supplemented with alpha-ketoisocaproate as well as with leucine, the transaminase catalyzing the last step in the leucine pathway was apparently normal in all strains. Therefore, the wild-type, auxotrophic, and several heterokaryotic strains were assayed for the activities of the other enzymes specific to leucine biosynthesis. Leu2 and Leu4 strains (complementation group I) lacked only alpha-isopropylmalate synthase activity; Leu3 and Leu6 strains (group III) lacked isopropylmalate isomerase activity; and Leu1 and Leu5 strains (group II) lacked beta-isopropylmalate dehydrogenase. Heterokaryons formed from leucine auxotrophs of different complementation groups had levels of activity for all three enzymes similar to those found in the wild-type strain. PMID- 16347074 TI - Temporal variation of denitrification activity in plant-covered, littoral sediment from lake hampen, denmark. AB - Diel and seasonal variations in denitrification were determined in a littoral lake sediment colonized by the perennial macrophyte Littorella uniflora (L.) Aschers. In the winter, the activity was low (5 mumol of N m h) and was restricted to the uppermost debris layer at a depth of 0 to 1 cm. By midsummer, the activity increased to 50 mumol of N m h and was found throughout the root zone to a depth of 10 cm. The root zone accounted for as much as 50 to 70% of the annual denitrification. A significant release of organic substrates from the roots seemed to determine the high activities of root zone denitrification in the summer. The denitrification in the surface layer and in the root zone formed two distinct activity zones in the summer, when the root zone also contained nitrification activity, as indicated from the accumulations of NO(3). Light conditions inhibited denitrification in both the surface layer and the upper part of the root zone, suggesting that a release of O(2) by benthic algae and from the roots of L. uniflora controlled a diel variation of denitrification. In midsummer, the rate of denitrification in both the surface layer and the upper part of the root zone was limited by NO(3). In the growth season, there was evidence for a significant population of denitrifiers closely associated with the root surface. PMID- 16347075 TI - A new sulfur-reducing, extremely thermophilic eubacterium from a submarine thermal vent. AB - A newly described bacterial isolate, designated strain NS-E, differs from presently known extremely thermophilic bacteria in various characteristics. It is a strictly heterotrophic eubacterium of marine origin and has a temperature range for growth of 50 to 95 degrees C with an optimum at 77 degrees C and a pH of 7.5. Its DNA base composition is 41.3 mol% guanine + cytosine. It is obligately anaerobic, utilizes various sugars as well as yeast extract, and reduces elemental sulfur facultatively to hydrogen sulfide. In 24-h cultures cell densities are up to fourfold higher in the presence than in the absence of elemental sulfur. Sulfide concentrations of 1.0 and 10.0 mM limit growth by 65 and 95%, respectively. Oxygen sensitivity is apparent only at or above that range of temperature at which growth occurs. PMID- 16347076 TI - Countermeasures to microbiofouling in simulated ocean thermal energy conversion heat exchangers with surface and deep ocean waters in hawaii. AB - Countermeasures to biofouling in simulated ocean thermal energy conversion heat exchangers have been studied in single-pass flow systems, using cold deep and warm surface ocean waters off the island of Hawaii. Manual brushing of the loops after free fouling periods removed most of the biofouling material. However, over a 2-year period a tenacious film formed. Daily free passage of sponge rubber balls through the tubing only removed the loose surface biofouling layer and was inadequate as a countermeasure in both titanium and aluminum alloy tubes. Chlorination at 0.05, 0.07, and 0.10 mg liter for 1 h day lowered biofouling rates. Only at 0.10 mg liter was chlorine adequate over a 1-year period to keep film formation and heat transfer resistance from rising above the maximum tolerated values. Lower chlorination regimens led to the buildup of uneven or patchy films which produced increased flow turbulence. The result was lower heat transfer resistance values which did not correlate with the amount of biofouling. Surfaces which were let foul and then treated with intermittent or continuous chlorination at 0.10 mg of chlorine or less per liter were only partially or unevenly cleaned, although heat transfer measurements did not indicate that fact. It took continuous chlorination at 0.25 mg liter to bring the heat transfer resistance to zero and eliminate the fouling layer. Biofouling in deep cold seawater was much slower than in the warm surface waters. Tubing in one stainless steel loop had a barely detectable fouling layer after 1 year in flow. With aluminum alloys sufficient corrosion and biofouling material accumulated to require that some fouling coutermeasure be used in long-term operation of an ocean thermal energy conversion plant. PMID- 16347077 TI - Automatic determination of bacterioplankton biomass by image analysis. AB - Image analysis was applied to epifluorescense microscopy of acridine orange stained plankton samples. A program was developed for discrimination and binary segmentation of digitized video images, taken by an ultrasensitive video camera mounted on the microscope. Cell volumes were estimated from area and perimeter of the objects in the binary image. The program was tested on fluorescent latex beads of known diameters. Biovolumes measured by image analysis were compared with directly determined carbon biomasses in batch cultures of estuarine and freshwater bacterioplankton. This calibration revealed an empirical conversion factor from biovolume to biomass of 0.35 pg of C mum (+/- 0.03 95% confidence limit). The deviation of this value from the normally used conversion factors of 0.086 to 0.121 pg of C mum is discussed. The described system was capable of measuring 250 cells within 10 min, providing estimates of cell number, mean cell volume, and biovolume with a precision of 5%. PMID- 16347078 TI - Effect of nitrate on biogenic sulfide production. AB - The addition of 59 mM nitrate inhibited biogenic sulfide production in dilute sewage sludge (10% [vol/vol]) amended with 20 mM sulfate and either acetate, glucose, or hydrogen as electron donors. Similar results were found when pond sediment or oil field brines served as the inoculum. Sulfide production was inhibited for periods of at least 6 months and was accompanied by the oxidation of resazurin from its colorless reduced state to its pink oxidized state. Lower amounts of nitrate (6 or 20 mM) and increased amounts of sewage sludge resulted in only transient inhibition of sulfide production. The addition of 156 mM sulfate to bottles with 59 mM nitrate and 10% (vol/vol) sewage sludge or pond sediment resulted in sulfide production. Nitrate, nitrite, and nitrous oxide were detected during periods where sulfide production was inhibited, whereas nitrate, nitrite, and nitrous oxide were below detectable levels at the time sulfide production began. The oxidation of resazurin was attributed to an increase in nitrous oxide which persisted in concentration of about 1.0 mM for up to 5 months. The numbers of sulfate-reducing organisms decreased from 10 CFU ml sludge to less than detectable levels after prolonged incubation of oxidized bottles. The addition of 10 mM glucose to oxidized bottles after 14.5 weeks of incubation resulted in rereduction of the resazurin and subsequent sulfide production. The prolonged inhibition of sulfide production was attributed to an increase in oxidation-reduction potential due to biogenic production of nitrous oxide, which appeared to have a cytotoxic effect on sulfate-reducing populations. PMID- 16347079 TI - Bradyrhizobium japonicum Serocluster 123 and Diversity among Member Isolates. AB - Diversity was examined within a group of 79 isolates of Bradyrhizobium japonicum reactive to fluorescent antibodies (FAs) prepared against B. japonicum USDA 123. Analyses were by means of cross-adsorbed FAs, bacteriophage typing, and endonuclease restriction digest patterns. Serogroups 127 and 129 shared antigenic determinants with serogroup 123 but not with each other. Bacteriophage and DNA digest patterns reflected more common features between serogroups 123 and 127 than between 123 and 129. Serogroups 129 and 122 showed FA cross-reactivity. The term serocluster was proposed to reflect interrelationships observed among the serogroups. PMID- 16347080 TI - Isolation of halotolerant, thermotolerant, facultative polymer-producing bacteria and characterization of the exopolymer. AB - Over 200 bacterial strains were selected for anaerobic growth at 50 degrees C and extracellular polysaccharide production in a sucrose-mineral salts medium with NaNO(3) and up to 10% NaCl. The predominant cell type was an encapsulated gram positive, motile, facultative sporeforming rod similar to Bacillus species. Strain SP018 grew and produced the polysaccharide on a variety of substrates at salinities up to 12% NaCl. Good polymer production only occurred anaerobically and was optimal between 4 and 10% NaCl. The ethanol-precipitated SP018 polymer was a charged heteropolysaccharide that contained glucose, mannose, arabinose, ribose, and low levels of allose and glucosamine. The SP018 polymer showed pseudoplastic behavior, was resistant to shearing, and had a higher viscosity at dilute concentrations and at elevated temperatures than xanthan gum. High-ionic strength solutions reversibly decreased the viscosity of SP018 polymer solutions. The bacterium and the associated polymer have many properties that make them potentially useful for in situ microbially enhanced oil recovery processes. PMID- 16347081 TI - Influence of External pH and Fermentation Products on Clostridium acetobutylicum Intracellular pH and Cellular Distribution of Fermentation Products. AB - Clostridium acetobutylicum ATCC 824 cells harvested from a phosphate-limited chemostat culture maintained at pH 4.5 had intracellular concentrations of acetate, butyrate, and butanol which were 13-, 7-, and 1.3-fold higher, respectively, than the corresponding extracellular concentrations. Cells from a culture grown at pH 6.5 had intracellular concentrations of acetate and butyrate which were only 2.2-fold higher than the respective external concentrations. The highest intracellular concentrations of these acids were attained at ca. pH 5.5. When cells were suspended in anaerobic citrate-phosphate buffer at pH 4.5, exogenous acetate and butyrate caused a concentration-dependent decrease in the intracellular pH, while butanol had relatively little effect until the external concentration reached 150 mM. Acetone had no effect at concentrations up to 200 mM. These data demonstrate that acetate and butyrate are concentrated within the cell under acidic conditions and thus tend to lower the intracellular pH. The high intracellular butyrate concentration presumably leads to induction of solvent production, thereby circumventing a decrease in the intracellular pH great enough to be deleterious to the cell. PMID- 16347082 TI - Stimulation of methanogenesis by aldicarb and several other N-methyl carbamate pesticides. AB - Aldicarb and several other N-methyl carbamate pesticides stimulated methane production in anaerobic salt marsh soils and organic-rich aquifer soils. Stimulation was biological and linearly related to the amount of carbamate added. Of the four carbamates studied, methomyl gave the greatest stimulation followed by carbaryl, aldicarb, and baygon. The percent conversions [(moles of CH(4) in excess of control/mole of carbamate added) x 100] for methomyl, carbaryl, aldicarb, and baygon were 88, 57, 40, and 11, respectively. Using aldicarb as a model carbamate, we found that monomethylamine (MA) accumulated in sediments as a result of aldicarb addition. MA arises from the N-methyl carbamoyl portion of the carbamates as a result of presumptive biological hydrolysis. MA levels decreased as CH(4) production was stimulated, and 2-bromoethane sulfonic acid (a specific inhibitor of mathanogenesis) partially inhibited the loss of MA. These findings suggest that N-methyl carbamates are readily hydrolyzed to MA in the presence of an active microbial population under anaerobic conditions and that methanogenesis is stimulated as a result of the consumption of MA by methanogenic bacteria. PMID- 16347083 TI - Mutants of Pachysolen tannophilus with Improved Production of Ethanol from d Xylose. AB - The conversion of d-xylose to ethanol by the yeast Pachysolen tannophilus is relatively inefficient in batch culture. The inefficiency has been attributed in part to concurrent utilization of ethanol in the presence of appreciable concentrations of d-xylose and to the formation of xylitol and other by-products. To increase the concentration of ethanol accumulated in batch cultures, UV induced mutants of P. tannophilus were selected on the basis of diminished growth on ethanol. Eleven independent mutant loci that conferred the ethanol-defective phenotype were identified. Three led to a greater yield and volumetric rate of production of ethanol than the wild type. One also produced less xylitol and was characterized by a deficiency in activity for malate dehydrogenase. PMID- 16347084 TI - Impact of storms on heterotrophic activity of epilimnetic bacteria in a southwestern reservoir. AB - The impact of storm conditions on the heterotrophic activity of planktonic bacteria in a southwestern reservoir was investigated. Storm events were considered as rainfall in excess of 2.5 cm in a 24-h period before sampling. Storm conditions stimulated heterotrophic activities and resulted in increased uptake rates and decreased turnover times of glutamate and acetate. Uptake rates were 45 to 75% faster immediately after storm conditions than they were during calm conditions. Activity levels appeared to return to prestorm levels within 48 h. Bacterial cell numbers did not change substantially during storm events. Cell specific activity indicated that increases in heterotrophic activity were the result of increased activity of individual cells. Light penetration, levels of particulate organic carbon, K(t) + S(n) values, and population levels of attached bacteria suggest that immediate sediment loading of the reservoir or increased substrate levels could not account for abrupt increases in heterotrophic activities. PMID- 16347085 TI - Conjugal Transfer of Bacteriophage Resistance Determinants on pTR2030 into Streptococcus cremoris Strains. AB - Agar surface conjugal matings were used to introduce heat-sensitive phage resistance (Hsp) determinants carried on the conjugal plasmid pTR2030 into Streptococcus cremoris KH, HP, 924, and TDM1. Lactose-fermenting (Lac) transconjugants were selected from matings of Lac variants of S. cremoris KH, HP, 924, and TDM1 with Streptococcus lactis ME2 or a high-frequency donor, S. lactis T-EK1 (pTR1040, Lac; pTR2030, Hsp). For all of the S. cremoris strains examined, select Lac transconjugants were completely resistant to plaquing by their homologous lytic phages. In all cases the plaquing efficiencies were less than 10. Acquisition of a 30-megadalton plasmid (pTR2030) in the S. cremoris phage resistant transconjugants was demonstrated by direct plasmid analysis, by hybridization with P-labeled probes, or by conjugal transfer of pTR2030 out of the phage-resistant transconjugants into a plasmid-cured recipient, S. lactis LM2302. Acid production, coagulation ability, and proteolytic activity of phage resistant transconjugants in milk were comparable to those of their phage sensitive parents. Further, S. cremoris phage-resistant transconjugants were not attacked by phage in starter culture activity tests, which included a 40 degrees C incubation period. The results demonstrated that phage resistance determinants on pTR2030 could be conjugally transferred to a variety of S. cremoris strains and confer resistance to phage under conditions encountered during cheese manufacture. Phage-resistant transconjugants of S. cremoris M43 and HP were also constructed without the use of antiblotic markers to select conjugal recipients from mating mixtures. PMID- 16347086 TI - Bacteriophage Resistance Conferred on Lactic Streptococci by the Conjugative Plasmid pTR2030: Effects on Small Isometric-, Large Isometric-, and Prolate Headed Phages. AB - A series of reactions between phages, sensitive hosts, and transconjugants where the sensitivity of small isometric-, large isometric-, and prolate-headed phages to pTR2030-induced phage resistance was evaluated in Streptococcus lactis and Streptococcus cremoris strains. Phage-resistant transconjugants were constructed in the desired host by conjugal transfer of lactose-fermenting ability (Lac, pTR1040) and phage resistance (Hsp, pTR2030) from S. lactis TEK1. S. lactis and S. cremoris transconjugants harboring pTR2030 were resistant to all small isometric-headed phages examined. In contrast, prolate- and large isometric headed phages were either not inhibited in the pTR2030 transconjugants or exhibited a reduction in plaque size without a reduction in the efficiency of plaquing. Small isometric-headed phages subject to pTR2030 induced inhibition shared no significant DNA homology with pTR2030, suggesting that phage immunity genes are not harbored on the plasmid or responsible for resistance. The general effectiveness of pTR2030 against small isometric-headed phages was highly significant since these are the phages which have been isolated most commonly from dairy fermentation plants. PMID- 16347087 TI - Thermal Tolerance of Zymomonas mobilis: Temperature-Induced Changes in Membrane Composition. AB - The membrane composition of Zymomonas mobilis changed dramatically in response to growth temperature. With increasing temperature, the proportion of vaccenic acid declined with an increase in myristic acid, the proportion of phosphatidylcholine and cardiolipin increased with decreases in phosphatidylethanolamine and phosphatidylglycerol, and the phospholipid/protein ratio of the membrane declined. These changes in membrane composition were correlated with changes in thermal tolerance and with changes in membrane fluidity. Cells grown at 20 degrees C were more sensitive to inactivation at 45 degrees C than were cells grown at 30 degrees C, as expected. However, cells grown at 41 degrees C (near the maximal growth temperature for Z. mobilis) were hypersensitive to thermal inactivation, suggesting that cells may be damaged during growth at this temperature. When cells were held at 45 degrees C, soluble proteins from cells grown at 41 degrees C were rapidly lost into the surrounding buffer in contrast to cells grown at lower temperatures. The synthesis of phospholipid-deficient membranes during growth at 41 degrees C was proposed as being responsible for this increased thermal sensitivity. PMID- 16347088 TI - Properties of a Clostridium thermocellum Endoglucanase Produced in Escherichia coli. AB - A cellulase gene of Clostridium thermocellum was transferred to Escherichia coli by molecular cloning with bacteriophage lambda and plasmid vectors and shown to be indentical with the celA gene. The celA gene product was purified from extracts of plasmid-bearing E. coli cells by heat treatment and chromatography on DEAE-Trisacryl. It was characterized as a thermophilic endo-beta-1,4-glucanase, the properties of which closely resemble those of endoglucanase A previously isolated from C. thermocellum supernatants. On sodium dodecyl sulfate polyacrylamide gel electrophoresis the enzyme purified from E. coli exhibited two protein bands with molecular weights of 49,000 and 52,000. It had a temperature optimum at 75 degrees C and was stable for several hours at 60 degrees C. Endoglucanase activity was optimal between pH 5.5 and 6.5. The enzyme was insensitive against end product inhibition by glucose and cellobiose and remarkably resistant to the denaturing effects of detergents and organic solvents. It was capable of degrading, in addition to cellulosic substrates, glucans with alternating beta-1,4 and beta-1,3 linkages such as barley beta glucan and lichenan. PMID- 16347089 TI - High-Efficiency Carbohydrate Fermentation to Ethanol at Temperatures above 40 degrees C by Kluyveromyces marxianus var. marxianus Isolated from Sugar Mills. AB - A number of yeast strains, isolated from sugar cane mills and identified as strains of Kluyveromyces marxianus var. marxianus, were examined for their ability to ferment glucose and cane syrup to ethanol at high temperatures. Several strains were capable of rapid fermentation at temperatures up to 47 degrees C. At 43 degrees C, >6% (wt/vol) ethanol was produced after 12 to 14 h of fermentation, concurrent with retention of high cell viability (>80%). Although the type strain (CBS 712) of K. marxianus var. marxianus produced up to 6% (wt/vol) ethanol at 43 degrees C, cell viability was low, 30 to 50%, and the fermentation time was 24 to 30 h. On the basis of currently available strains, we suggest that it may be possible by genetic engineering to construct yeasts capable of fermenting carbohydrates at temperatures close to 50 degrees C to produce 10 to 15% (wt/vol) ethanol in 12 to 18 h with retention of cell viability. PMID- 16347090 TI - Chemical Composition of the Edible Mushroom Pleurotus ostreatus Produced by Fermentation. AB - Pleurotus ostreatus ;Florida' was grown in submerged liquid culture. The biomass yield of the fungus, grown for 3 days in 2-liter fermentors, where the mycelial pellets measuring 5 mm in diameter were formed, was 11.7 g (dry weight)/liter. Comparing the chemical constituents of fruiting bodies produced on cotton straw and mycelial pellets revealed several similarities in total nitrogen, protein, glycogen, fatty acids, RNA, and ash content. Differences were observed in the contents of six amino acids. Although the total fatty acid content was similar, there were more saturated fatty acids in the mycelium. Cell wall composition, typical for basidiomycetes, was observed in both mycelium and fruiting bodies, with laminarin as the main polymer. PMID- 16347091 TI - Resistance to In Vitro Restriction of DNA from Lactic Streptococcal Bacteriophage c6A. AB - DNA isolated from streptococcal bacteriophage c6A was cut only infrequently by many restriction endonucleases. Fragments of c6A DNA cloned in Escherichia coli plasmids were similarly resistant to cleavage. We conclude that the low frequency of cleavage is due to an unusually low number of restriction enzyme recognition sequences in c6A DNA. PMID- 16347092 TI - Isolation of Endoglucanase Genes from Pseudomonas fluorescens subsp. cellulosa and a Pseudomonas sp. AB - Endoglucanase genes from Pseudomonas fluorescens subsp. cellulosa and Pseudomonas sp. were cloned and characterized. DNA hybridization studies showed that these genes are homologous and that each species has one copy of the gene per genome. The DNA fragment from Pseudomonas sp. codes for, at most, a 23-kilodalton endoglucanase. PMID- 16347093 TI - Protoplast Transformation in Coryneform Bacteria and Introduction of an alpha Amylase Gene from Bacillus amyloliquefaciens into Brevibacterium lactofermentum. AB - [This corrects the article on p. 638 in vol. 51.]. PMID- 16347094 TI - Evaluation of different assay systems for identification of environmental Aeromonas strains. AB - [This corrects the article on p. 655 in vol. 51.]. PMID- 16347095 TI - Sensitivity of the Entomogenous Fungus Beauveria bassiana to Selected Plant Growth Regulators and Spray Additives. AB - Mefluidide was the only one of four plant growth regulators that caused little to no significant inhibition of in vitro germination and growth of the entomogenous fungus Beauveria bassiana. Silaid, paclobutrazol, and flurprimidol significantly inhibited germination and growth. Mortality of fall armyworm, Spodoptera frugiperda, resulting from B. bassiana was significantly reduced when larvae were exposed to conidia plus soil treated with paclobutrazol. Larval mortality resulting from conidia plus soil treated with mefluidide did not differ significantly from mortality resulting from untreated conidia. Triton CS-7 was the only one of eight spray adjuvants that significantly inhibited germination of B. bassiana conidia. PMID- 16347096 TI - Use of metabolic inhibitors to estimate protozooplankton grazing and bacterial production in a monomictic eutrophic lake with an anaerobic hypolimnion. AB - Inhibitors of eucaryotes (cycloheximide and amphotericin B) and procaryotes (penicillin and chloramphenicol) were used to estimate bacterivory and bacterial production in a eutrophic lake. Bacterial production appeared to be slightly greater than protozoan grazing in the aerobic waters of Lake Oglethorpe. Use of penicillin and cycloheximide yielded inconsistent results in anaerobic water and in aerobic water when bacterial production was low. Production measured by inhibiting eucaryotes with cycloheximide did not always agree with [H]thymidine estimates or differential filtration methods. Laboratory experiments showed that several common freshwater protozoans continued to swim and ingest bacterium-size latex beads in the presence of the eucaryote inhibitor. Penicillin also affected grazing rates of some ciliates. We recommend that caution and a corroborating method be used when estimating ecologically important parameters with specific inhibitors. PMID- 16347097 TI - Bacterial inhibitors in lake water. AB - The populations of six bacterial genera fell rapidly after their addition to sterile lake water but not after their addition to buffer. The decline in numbers of two species that were studied further, Klebsiella pneumoniae and Micrococcus flavus, occurred even when the buffer was added to sterile lake water. The inhibition of K. pneumoniae by substances in lake water varied with the season of the year, and the rate and extent of decline of both species were different in sterile samples of different lakes. The extent of reduction in the density of K. pneumoniae was independent of initial population size and was diminished by the addition of 10 mug of glucose per ml of lake water. The toxin was removed from lake water by dialysis and by a cation-exchange resin but not by an anion exchange resin, and it was destroyed by heating. The inhibition of K. pneumoniae was not evident in lake water buffered at a pH value above 8.0. We suggest that toxins may be important in determining the composition of the bacterial community of lakes. PMID- 16347098 TI - Production and Use of Monoclonal Antibodies for Identification of Strains of Rhizobium trifolii. AB - We produced a monoclonal antibody against Rhizobium trifolii 162x95. This antibody in cell culture supernatant was used in an indirect enzyme-linked immunosorbent assay to differentiate strain 162x95 from naturalized strains in the Appalachian region. Nodules crushed in 0.1 to 0.2 ml of phosphate-buffered saline and used to charge enzyme-linked immunosorbent assay plates gave strong absorbance readings. Heat-inactivated and noninactivated portions of 162x95 cultures were strongly reactive, indicating that the antigen is probably a carbohydrate. Of 10 strains from California, where 162x95 was isolated, 6 strongly cross-reacted with the antibody. The cellular protein patterns in a sodium dodecyl sulfate-polyacrylamide gradient gel of cross-reactive strains were essentially identical. A Western blot analysis indicated that the antibody was against a 19.8-kilodalton band. The Western blot analysis also revealed that the polyvalent antiserum contained other strongly reacting antibodies with molecular weights of approximately 20,000, indicating the possibility that other monoclonal antibodies to detect strain 162x95 may be produced. However, the available antibody has been shown to be useful for short-term experiments. Based upon protein profiles and immunological reactions, there are 4 or 5 California strains rather than 10. PMID- 16347099 TI - Biochemical Aspects of Fumaric Acid Accumulation by Rhizopus arrhizus. AB - The accumulation and excretion of fumaric acid, and to a lesser extent malic and succinic acids, by Rhizopus arrhizus occurs under aerobic conditions in a high glucose medium containing a limiting amount of nitrogen and a neutralizing agent (CaCO(3)). An overall four-carbon dicarboxylic acid molar yield of up to 145% (moles of acid produced per mole of glucose utilized) is obtained after incubation for 4 to 5 days. Evidence is presented that fumarate is synthesized from pyruvate via a carboxylation reaction yielding oxaloacetate, which is then converted to malate and further on to fumarate via the reductive reactions of the tricarboxylic acid cycle. The possible formation of fumarate from the normal (oxidative) operation of the tricarboxylic acid cycle was not excluded by the data. Yield, C nuclear magnetic resonance, and enzymatic activity studies were carried out in a strain of R. arrhizus which produces high levels of fumarate from glucose and carbonate. The observed high fumarate molar yield (greater than 100%) can therefore be explained in terms of the carboxylation of pyruvate and the operation of the reductive reactions of the tricarboxylic acid cycle under aerobic conditions. PMID- 16347100 TI - Role of Pili (Fimbriae) in Attachment of Bradyrhizobium japonicum to Soybean Roots. AB - Pili (fimbriae) were observed on cells of each of the five strains of Bradyrhizobium japonicum and the one strain of Rhizobium trifolii examined. Pili on B. japonicum were about 4 nm in diameter and polarly expressed. Piliated cells were estimated by transmission electron microscopy and hydrophobic attachment to polystyrene to constitute only a small percentage of the total population. The proportion of piliated cells in these populations was dependent on culture age in some strains. Piliated B. japonicum cells were selectively and quantitatively removed from suspension when cultures were incubated with either soybean roots or hydrophobic plastic surfaces, indicating that pili were involved in the attachment of the bacteria to these surfaces. Pili from B. japonicum 110 ARS were purified and found to have a subunit molecular weight of approximately 21,000. Treatment of B. japonicum suspensions with antiserum against the isolated pili reduced attachment to soybean roots by about 90% and nodulation by about 80%. Pili appear to be important mediators of attachment of B. japonicum to soybean roots under the conditions examined. PMID- 16347101 TI - Use of a nylon manufacturing waste as an industrial fermentation substrate. AB - Nonvolatile residue (NVR), a waste stream from the manufacture of nylon 6'6', contains mainly small carboxylic acids and alcohols, making it a potential fermentation substrate. Above a concentration of 1.3% (wt/vol), NVR inhibited the growth of all microorganisms tested. The most inhibitory of the major NVR components were the monocarboxylic acids (C(4) to C(6)) and epsilon-caprolactone. The inhibitory effects of NVR could be avoided by using a carbon-limited chemostat. Microorganisms were found that could use all of the major NVR components as carbon and energy sources. One such organism, Pseudomonas cepacia, was grown in a carbon-limited chemostat with a medium feed concentration of 20.5 g of NVR liter. At a dilution rate of 0.14 h the yield of biomass (Y(x/s), where x is biomass produced and s is substrate used) from NVR was 18% (neglecting the water content of NVR). It was concluded that NVR would be a suitable carbon source for certain industrial fermentation processes such as the production of poly-beta-hydroxybutyric acid. PMID- 16347102 TI - Isolation and Characterization of an Fe(III)-Chelating Compound Produced by Pseudomonas syringae. AB - The phytopathogenic bacterium Pseudomonas syringae produces a fluorescent pigment when it is grown in iron-deficient media. This pigment forms a very stable Fe(III) complex that was purified in this form by using a novel procedure based on ultrafiltration and column chromatography. The Fe(III) complex has a molecular weight of 1,100 and contains 1 mol of Fe(III). The pigment is composed of an amino acid moiety with three threonines, three serines, one lysine, delta-N hydroxyornithine, and a quinoline-type fluorescent chromophore. These features and its stability constant (in the range of 10) suggest that the fluorescent pigment of P. syringae is related to the siderophores produced by another Pseudomonas species. PMID- 16347103 TI - Microoxic-Anoxic Niche of Beggiatoa spp.: Microelectrode Survey of Marine and Freshwater Strains. AB - Beggiatoa spp. grow optimally in media containing opposed gradients of oxygen and soluble sulfide, although some strains also require an organic substrate. By using microelectrodes, we characterized oxygen and sulfide gradients during their initial development in uninoculated media and in cultures of marine and freshwater strains. In gradient media, Beggiatoa strains always grew some distance below the air/agar interface as a dense "plate" of constantly gliding filaments with sharply demarcated upper and lower boundaries. Within established plates, the maximum oxygen partial pressure was 0.6 to 6.0% of air saturation and not significantly lower if filaments were fixing nitrogen. Oxygen penetrated only 100 to 300 mum into the plate, and the anoxic fraction increased from less than 10% to approximately 90% during later stages of growth. For lithoautotrophically grown marine strains, the linearity of the oxygen profile above the plate plus its drop to zero therein indicated that oxygen uptake for the entire tube occurred only within the Beggiatoa plate. Consequently, oxygen consumption could be predicted solely from the distance between the air/agar interface and the top of a plate, given the diffusion coefficient for oxygen. By contrast, for freshwater strains grown heterotrophically (with sulfide also in the medium), oxygen profiles were frequently nonlinear because of nonbiological reaction with sulfide which had diffused past the aggregated filaments. For all strains tested, microoxic aggregation also occurred in the absence of sulfide, apparently reflecting a step-up phobic response to oxygen. PMID- 16347104 TI - Nutritional Factors Affecting the Ratio of Solvents Produced by Clostridium acetobutylicum. AB - Fermentation of whey by Clostridium acetobutylicum yielded butanol and acetone in a ratio of approximately 100:1. This ratio amounted to only 2:1 in synthetic media with glucose, lactose, or glucose plus galactose as substrates. Removal of citrate from whey and addition of minerals resulted in an increase in the amount of acetone produced. Experiments carried out in a chemostat with a low-phosphate synthetic medium revealed that the butanol/acetone ratio could be increased from 2:1 to 3.8:1 by cofermentation of l-lactate and from 2:1 to 8:1 by iron limitation. The performance of the fermentation in a low-iron glucose medium above pH 5.1 yielded l-lactate as the main product. PMID- 16347105 TI - Life Cycles in the Methanogenic Archaebacterium Methanosarcina mazei. AB - Methanosarcina mazei S6 and LYC were used to study the structure and differentiation of the aggregating methanogens. Cultures harvested under various conditions are described at the ultrastructural level. Cells of strain S6 are enclosed by a layer 12 nm thick in contact with the plasma membrane. In sarcinal colonies, cells are held in close association by a fibrous matrix up to 60 nm thick. Colony maturation was examined in strain S6 over a period of 1 year. Changes occurred in the shape and staining of individual cells. Also, various inclusion bodies were observed that either persist throughout colony maturation or are only found at certain growth stages. Two types of cores that are composed of double membranes in M. mazei S6 are described. One has a 90-nm diameter and contains electron-dense granules similar to those found in the cytoplasm. The other core type does not contain granules, is more numerous, and is found in older cultures. Two life cycles are described for M. mazei based on electron microscope examinations. A complex life cycle involving the release of single cells is described with two variations for strains S6 and LYC. When released cells of strain S6 are placed in fresh medium they can repeat the cycle. In addition, a limited cycle is described for both strains of M. mazei. This limited cycle contains the only sarcinal morphotypes observed in M. barkeri. When M. mazei S6 remains in the limited cycle and does not disaggregate in stationary phase, several types of possible resting forms are found. PMID- 16347106 TI - Seasonal population changes and characterization of ice-nucleating bacteria in farm fields of central alberta. AB - During the summer of 1983 in central Alberta, changes in the bacterial population inhabiting the leaves of field beans (Phaseolus vulgaris L.) and canola (Brassica napus L. Altex) were studied to determine if ice-nucleating bacteria were present on these plants. Three colony types (white, yellow, and peach-colored) were found on field beans and canola leaves. Approximately 25% of the isolates from the white colony group, which dominated the population, were ice-nucleating bacteria. No ice-nucleating bacteria were present on canola leaves. Out of a total of 76 ice-nucleating bacteria isolated, 5 representative cultures were characterized in detail and identified as Pseudomonas fluorescens. The fatty acid composition of these cultures was essentially identical to that of typical P. fluorescens cultures and was altered by varying the growth temperature from 10 to 30 degrees C. PMID- 16347107 TI - Culture Conditions for Production of Thermostable Amylase by Bacillus stearothermophilus. AB - Bacillus stearothermophilus grew better on complex and semisynthetic medium than on synthetic medium supplemented with amino acids. Amylase production on the complex medium containing beef extract or corn steep liquor was higher than on semisynthetic medium containing peptone (0.4%). The synthetic medium, however, did not provide a good yield of extracellular amylase. Among the carbohydrates which favored the production of amylase are, in order starch > dextrin > glycogen > cellobiose > maltohexaose-maltopeptaose > maltotetraose and maltotriose. The monosaccharides repressed the enzyme production, whereas inositol and d-sorbitol favored amylase production. Organic and inorganic salts increased amylase production in the order of KCI > sodium malate > potassium succinate, while the yield was comparatively lower with other organic salts of Na and K. Amino acids, in particular isoleucine, cysteine, phenylalanine, and aspartic acids, were found to be vital for amylase synthesis. Medium containing CaCl(2) 2H(2)O enhanced amylase production over that on Ca -deficient medium. The detergents Tween-80 and Triton X-100 increased biomass but significantly suppressed amylase synthesis. The amylase powder obtained from the culture filtrate by prechilled acetone treatment was stable over a wide pH range and liquefied thick starch slurries at 80 degrees C. The crude amylase, after (NH(4))(2)SO(4) fractionation, had an activity of 210.6 U mg. The optimum temperature and pH of the enzyme were found to be 82 degrees C and 6.9, respectively. Ca was required for the thermostability of the enzyme preparation. PMID- 16347108 TI - Characterization, Biosynthesis, and Regulation of Granulose in Clostridium acetobutylicum. AB - Synthesis of granulose was investigated in 15 solvent-producing Clostridium strains. Only one of the strains did not produce granulose. The structure of granulose in Clostridium acetobutylicum P262 consisted of a high-molecular-weight polyglucan containing only (1-->4) linked d-glucopyranose units. Biosynthesis of granulose in C. acetobutylicum P262 was dependent on ADPglucose pyrophosphorylase, and granulose synthase and mutants defective in granulose accumulation lacked either one or both enzyme activities. Granulose-positive revertants exhibited both enzyme activities. ADPglucose pyrophosphorylase and granulose synthase were not subject to allosteric control by metabolites. Granulose accumulation and the biosynthetic enzyme activities were initiated immediately before the pH breakpoint and were detected in cells only at the end of the exponential growth phase. Granulose accumulation did not occur under conditions of nitrogen limitation, excess carbon, or excess energy. PMID- 16347109 TI - Incorporation of Oxygen from Water into Toluene and Benzene during Anaerobic Fermentative Transformation. AB - Toluene and benzene were anaerobically transformed and eventually mineralized in mixed methanogenic cultures. However, the source of oxygen for the initial oxidation step had been unknown, owing to the presence of both methanol and water. No exogenous electron acceptors other than carbon dioxide, toluene, and benzene were present in the defined mineral medium. Through the use of O-labeled water, the oxygen incorporated into the monoaromatic compounds was shown to come from water. The cresol from the toluene and the phenol from the benzene contained up to 8% O label after incubation in 9% O-labeled medium. Gas chromatography-mass spectrometry was used to detect the O-labeled aromatic metabolites. PMID- 16347110 TI - Oxidation of chloroform in an aerobic soil exposed to natural gas. AB - Acclimation of a sandy soil to an air-natural gas mixture stimulated the biological oxidation of chloroform to carbon dioxide. Acetylene and methane inhibited chloroform oxidation. Chloroform oxidation continued up to 31 days in the absence of methane. Chloroform oxidation rates increased at chloroform concentrations up to 5 mug g of soil. PMID- 16347111 TI - Effect of 3-phenylpropanoic Acid on growth of and cellulose utilization by cellulolytic ruminal bacteria. AB - The growth of several cellulolytic species of ruminal bacteria was measured in media containing either cellobiose or cellulose as the energy source and with or without added 3-phenylpropanoic acid (PPA). With Ruminoccoccus albus 7 and 8, the addition of PPA greatly enhanced the rate of cellulose utilization but had little effect on the rate of growth when cellobiose was the energy source. Comparative rates of growth obtained on either cellobiose or cellulose for Ruminococcus flavefaciens FD1 or C94 and Butyrivibrio fibrisolvens 12, 49, or A38 were similar regardless of the PPA content of the growth medium. PMID- 16347112 TI - Agrocin-Producing Agrobacterium tumefaciens Strain Active against Grapevine Isolates. AB - Agrobacterium tumefaciens J73, a biotype 2 strain harboring a nopaline Ti plasmid, was found to produce an agrocin active against a broad range of A. tumefaciens strains, including grapevine isolates. Sensitivity to J73 is not encoded by a Ti plasmid. Optimal conditions for the production of the agrocin were determined. PMID- 16347113 TI - Hydrophobic interactions involved in attachment of a baculovirus to hydrophobic surfaces. AB - The hydrophobic interactions of Trichoplusia ni nuclear polyhedrosis virus were characterized by hydrophobic interaction chromatography. The determination of the hydrophobic force and some of the factors that influence its size is discussed in relation to the attachment to leaf surfaces of polyhedra during their use as biological control agents against insect pests. PMID- 16347114 TI - Carbon and nitrogen content of natural planktonic bacteria. AB - A method of estimating carbon and nitrogen content per unit of natural bacterial cell volume was developed. This method is based on the difference in the retentiveness of bacteria between two kinds of glass fiber filter, GF/C and GF/F (Whatman, Inc., Clifton, N.J.). Biovolume and biomass (carbon and nitrogen content) of bacteria which passed through the GF/C but not the GF/F filter were estimated with an epifluorescence microscopy and a CHN analyzer, respectively. From seasonal determinations of natural planktonic bacteria in epilimnetic waters of a mesotrophic lake, the conversion factors of 106 fg of C/mum and 25 fg of N/mum were derived as average values. By using these values, the contribution of bacteria to the biomass of lake plankton is discussed. PMID- 16347115 TI - Purification and Properties of a Novel Xanthan Depolymerase from a Salt-Tolerant Bacterial Culture, HD1. AB - A novel xanthan depolymerase (endo-beta-1,4-glucanase) was isolated from a salt tolerant bacteria culture (HD1) grown on xanthan. The depolymerase was purified 55-fold through chromatography on ion-exchange and molecular sieve columns, including high-performance liquid chromatography. The purified enzyme fraction was homogeneous as judged by polyacrylamide gel electrophoresis. The molecular weight of this enzyme is 60,000. Optimum pH and temperature for xanthan depolymerase activity were around 5 and 30 to 35 degrees C, respectively. The enzyme was not stable at a temperature higher than 45 degrees C. The activation energy calculated from an Arrhenius plot was 6.40 kcal (26.78 kJ). The enzyme molecule contains no sugar moiety. The amino acid composition of the enzyme protein was determined. Xanthan depolymerase cleaves the endo-beta-1,4-glucosidic linkage of the xanthan molecule, freeing reducing groups of some sugars and decreasing viscosity of the polymer solution. Only the backbones of beta-1,4 linked glucans with side chains or other substituents were cleaved. No monosaccharide was produced by the action of this enzyme. The oligosac-charide(s) in the low-molecular weight fraction consisted of 15 to 58 monosaccharide units. The enzymic reaction resulted in the decrease in weight-average molecular weight of xanthan from 6.5 x 10 to 8.0 x 10 in 0.5 h. This enzyme alone could not degrade xanthan to a single or multiple pentasaccharide unit(s). Results suggest that there may be regions inside the xanthan molecule that are susceptible to the attack of this enzyme. Xanthan depolymerase activity was not inhibited by many chemicals, including thiols, antioxidants, chlorinated hydrocarbons, metal chelating agents, and inorganic compounds, except ferric chloride and arsenomolybdate. Many biocides were tested and found not to be inhibitory. Conditions used in enhanced oil recovery operations, i.e., the presence of formaldehyde, Na(2)S(2)O(4), 2,2-dibromo-3-nitrilopropionamide, and an anaerobic environment, did not inhibit xanthan depolymerase activity. PMID- 16347116 TI - Use of genetically marked minicells as a probe in measurement of predation on bacteria in aquatic environments. AB - Minicells produced by Escherichia coli M2141 were used as probes to measure predation on pelagic bacteria in situ. The minicells, labeled with [S]methionine in one specific protein, were shown to disappear in the presence of a microflagellate (Ochromonas sp.), as seen by a decrease in the amount of labeled marker protein with time. Incubation in filtered (pore size, 0.2 mum) and autoclaved seawater did not affect the amount of labeled marker protein in the minicell. The generation time of flagellates feeding on minicells was determined to be similar to that found for flagellates grown on seawater bacteria or living E. coli NC3. Data indicate that minicells are seen as true food particles by the flagellates. The minicell probe was used in recapture experiments, in which predation in situ on pelagic bacteria was demonstrated. The rate of bacterial production showed a clear covariation with the rate of predation, both in different sea areas and in depth profiles. The obtained results (11 field experiments) showed that the rate of predation, on average, accounts for the consumption of 62% of the bacteria produced. PMID- 16347117 TI - Analysis of the Symbiotic Performance of Bradyrhizobium japonicum USDA 110 and Its Derivative I-110 and Discovery of a New Mannitol-Utilizing, Nitrogen-Fixing USDA 110 Derivative. AB - Previously, Bradyrhizobium japonicum USDA 110 was shown to contain colony morphology variants which differed in nitrogen-fixing ability. Mannitol-utilizing derivatives L1-110 and L2-110 have been shown to be devoid of symbiotic nitrogen fixation ability, and non-mannitol-utilizing derivatives I-110 and S-110 have been shown to be efficient at nitrogen fixation. The objectives of this study were to determine the effect of media carbon sources on the symbiotic N(2)-fixing ability of strain USDA 110 and to compare the effectiveness of strain USDA 110 and derivative I-110. Based on acetylene reduction activity and the nitrogen content of 41-day-old soybean plants, neither derivative I-110 nor cultures of USDA 110 grown in media favoring non-mannitol-using derivatives had symbiotic nitrogen fixation that was statistically superior to that of cultures of USDA 110 grown in media favoring mannitol-using derivatives. In another experiment 200 individual nodules formed by strain USDA 110 grown in yeast extract gluconate were screened for colony morphology of occupying variant(s) and acetylene reduction activity. Nodules occupied by mannitol-using derivatives (large colony type on 0.1% yeast extract-0.05% K(2)HPO(4)-0.08% MgSO(4) . 7H(2)O-0.02% NaCl 0.001% FeCl(3) . 6H(2)O [pH 6.7] with 1% mannitol [YEM] plates) had a mean acetylene reduction activity equal to that of nodules occupied by non-mannitol using derivatives (small colony type on YEM plates). A total of 20 large colonial derivatives and 10 small colonial derivatives (I-110-like) were isolated and purified by repeated culture in YEM and YEG (same as YEM except 1% gluconate instead of 1% mannitol) media, respectively, followed by dilution in solutions containing 0.05% Tween 40. After 25 days of growth, soybean plants inoculated with the large colony isolates had mean whole-plant acetylene reduction activity, whole-plant dry weight, and whole-plant nitrogen contents equal to or better than those of plants inoculated with either the small colony isolates (I-110-like) or the I-110 (non-mannitol-using) derivative. Hence, the existence of a mannitol utilizing derivative that fixes nitrogen in a culture of strain USDA 110 obtained from the U.S. Department of Agriculture, Beltsville, Md., was established. This new USDA 110 derivative was designated as MN-110 because it was a mannitol utilizing nitrogen-fixing USDA 110 derivative. This derivative was morphologically indistinguishable from the non-nitrogen-fixing derivative L2-110 found in cultures obtained earlier from the U.S. Department of Agriculture, Beltsville. DNA-DNA homology and restriction enzyme analyses indicated that MN 110 is genetically related to other USDA 110 derivatives that have been characterized previously. PMID- 16347118 TI - Characterization of a Mannitol-Utilizing, Nitrogen-Fixing Bradyrhizobium japonicum USDA 110 Derivative. AB - We have isolated a colonial derivative of Bradyrhizobium japonicum USDA 110 (designated MN-110) that is both mannitol utilizing and N(2) fixing. Derivative MN-110 showed growth on mannitol and glucose similar to that of non-N(2)-fixing, mannitol-utilizing L2-110. Derivative MN-110 showed high constitutive and induced d-mannitol dehydrogenase activity (similar to L2-110) relative to N(2)-fixing, non-mannitol-utilizing I-110. Hybridization to EcoRI and HindIII total DNA digests with cloned USDA 110 nif DK and nif H genes revealed similar patterns for non-N(2)-fixing mannitol-utilizing derivative L1-110 and derivative MN-110. Symbiotic tests with soybean cultivars Ransom and Lee indicate MN-110 to be a superior N(2)-fixing derivative compared with derivative I-110 and the parent strain USDA 110. However, these differences were not revealed when comparing 28 day-old soybean-B. japonicum associations but were apparent in 49-day-old associations. It was apparent from this work that mannitol utilization was not necessarily correlated to symbiotic effectiveness in B. japonicum and that gene rearrangements were not responsible for differences in N(2) fixation between L1 110 or L2-110 and MN-110. PMID- 16347119 TI - Intracellular Conditions Required for Initiation of Solvent Production by Clostridium acetobutylicum. AB - We investigated the intracellular physiological conditions associated with the induction of butanol-producing enzymes in Clostridium acetobutylicum. During the acidogenic phase of growth, the internal pH decreased in parallel with the decrease in the external pH, but the internal pH did not go below 5.5 throughout batch growth. Butanol was found to dissipate the proton motive force of fermenting C. acetobutylicum cells by decreasing the transmembrane pH gradient, whereas the membrane potential was affected only slightly. In growing cells, the switch from acid to solvent production occurred when the internal undissociated butyric acid concentration reached 13 mM and the total intracellular undissociated acid concentration (acetic plus butyric acids) was at least 40 to 45 mM. Similar values were obtained when cultures were supplemented with 50 mM butyric acid initially or when a phosphate-buffered medium was used instead of an acetate-buffered medium. To measure the induction of the enzymes involved in solvent synthesis, we determined the rates of conversion of butyrate to butanol in growing cells. The rate of butanol formation reached a maximum in the mid solvent phase, when the butanol concentration was 50 mM. Although more solvent accumulated later, de novo enzyme synthesis decreased and then ceased. PMID- 16347120 TI - Demethylation of Veratrole by Cytochrome P-450 in Streptomyces setonii. AB - The actinomycete Streptomyces setonii 75Vi2 demethylates vanillic acid and guaiacol to protocatechuic acid and catechol, respectively, and then metabolizes the products by the beta-ketoadipate pathway. UV spectroscopy showed that this strain could also metabolize veratrole (1,2-dimethoxybenzene). When grown in veratrole-containing media supplemented with 2,2'-dipyridyl to inhibit cleavage of the aromatic ring, S. setonii accumulated catechol, which was detected by both liquid chromatography and gas chromatography. Reduced cell extracts from veratrole-grown cultures, but not sodium succinate-grown cultures, produced a carbon monoxide difference spectrum with a peak at 450 nm that indicated the presence of soluble cytochrome P-450. Addition of veratrole or guaiacol to oxidized cell extracts from veratrole-grown cultures produced difference spectra that indicated that these compounds were substrates for cytochrome P-450. My results suggest that S. setonii produces a cytochrome P-450 that is involved in the demethylation of veratrole and guaiacol to catechol, which is then catabolized by the beta-ketoadipate pathway. PMID- 16347121 TI - Growth Pattern and Yield of a Chemoautotrophic Beggiatoa sp. in Oxygen-Sulfide Microgradients. AB - Recently developed techniques involving opposed, gel-stabilized gradients of O(2) and H(2)S permit cultivation of a marine Beggiatoa strain as a chemolithoautotroph which uses gliding motility to precisely track the interface between H(2)S and O(2). In the current study with microelectrodes, vertical profiles of H(2), O(2), and pH were measured in replicate cultures grown for various intervals. After an initial period of exponential biomass increase (doubling time, 11 h), linear growth prevailed throughout much of the time course. This H(2)S-limited growth was followed by a transition to stationary phase when the declining H(2)S flux was sufficient only to supply maintenance energy. During late-exponential and linear growth phases, the Beggiatoa sp. consumed a constant 0.6 mol of H(2)S for each 1.0 mol of O(2), the ratio anticipated for balanced lithoautotrophic growth at the expense of complete oxidation of H(2)S to SO(4). Over the entire range of conditions studied, this consumption ratio varied by approximately twofold. By measuring the extent to which the presence of the bacterial plate diminished the overlap of O(2) and H(2)S, we demonstrated that oxidation of H(2)S by Beggiatoa sp. is approximately 3 orders of magnitude faster than spontaneous chemical oxidation. By integrating sulfide profiles and comparing sulfide consumed with biomass produced, a growth yield of 8.4 g (dry weight) mol of H(2)S was computed. This is higher than that found for sulfide-grown thiobacilli, indicating very efficient growth of Beggiatoa sp. as a chemoautotroph. The methods used here offer a unique opportunity to determine the yield of H(2)S-oxidizing chemolithoautotrophs while avoiding several problems inherent in the use of homogeneous liquid culture. Finally, by monitoring time-dependent formation of H(2)S profiles under anoxic conditions, we demonstrate a method for calculating the molecular diffusion coefficient of soluble substrates in gel-stabilized media. PMID- 16347122 TI - Susceptibility of Agrobacterium tumefaciens Strains to Two Agrocin-Producing Agrobacterium Strains. AB - Sixty-five strains and isolates of Agrobacterium tumefaciens representing each of the known biotypes, were tested for in vitro and in vivo susceptibility to the agrocin-producing strains Agrobacterium radiobacter 84 and A. tumefaciens D286. No biotype 3 strain was susceptible to the effects of either of the agrocinogenic strains in vitro. On datura and tobacco, the best inhibition of tumor formation was obtained when the agrocinogenic strains were applied to wounds 24 h before the pathogens and by the concomitant application of agrocin producer and pathogen at cell ratios of 10:1 or 3:1; inhibition of infection tended to decrease progressively as the cell ratio decreased from 10:1 to 3:1 to 1:1. Generally, strain 84 was superior to D286 in inhibiting tumor formation. A combined cell suspension of 84 and D286 was as effective as 84 alone. The overall pattern of inhibition of tumor formation by biotype 1 and 2 pathogens resistant to the agrocinogenic strains in vitro was similar to that obtained with strains that were susceptible in vitro. PMID- 16347123 TI - Ultrastructural Aspects of Wood Delignification by Phlebia (Merulius) tremellosus. AB - Wood from aspen and birch that had been decayed for 12 weeks by Phlebia tremellosus had averages of 30 and 31% weight loss, respectively, and 70% lignin loss. Digestibility increased from averages of 21 and 13% for sound aspen and birch to 54 and 51% for decayed aspen and birch. Individual wood sugar analyses of decayed birch blocks indicated an average loss of 10% glucose, 45% xylose, and 19% mannose. Micromorphological studies demonstrated the removal of middle lamellae and separation of cells. Vessels also separated at perforation plates. Electron microscopy with OsO(4)-glutaraldehyde-fixed and KMnO(4)-fixed wood showed that lignin was progressively removed first from the secondary cell wall layers, beginning at the lumen surface, and later from the compound middle lamella. Extensive degradation of lignin was found throughout the secondary wall and middle lamella region between cells. In cells with advanced decay, the middle lamella between cells was completely degraded, but cell corner regions remained. PMID- 16347124 TI - Effects of pH on Lignin and Cellulose Degradation by Streptomyces viridosporus. AB - Lignocellulose degradation by Streptomyces viridosporus results in the oxidative depolymerization of lignin and the production of a water-soluble lignin polymer, acid-precipitable polymeric lignin (APPL). The effects of the culture pH on lignin and cellulose metabolism and APPL production by S. viridosporus are reported. Dry, ground, hot-water-extracted corn (Zea mays) lignocellulose was autoclaved in 1-liter reagent bottles (5 g per bottle) and inoculated with 50-ml volumes of S. viridosporus cells suspended in buffers of specific pH (pH 6.0 to 9.2 at 0.4 pH unit intervals). Four replicates of inoculated cultures and of uninoculated controls at each pH were incubated as solid-state fermentations at 37 degrees C. After 6 weeks of incubation the percent loss of lignocellulose, lignin, and carbohydrate and the amount of APPL produced were determined for each replicate. Optimal lignocellulose degradation, as shown by substrate weight loss, was observed in the pH range of 8.4 to 8.8. Only minor differences were seen in the Klason lignin, carbohydrate, protein, and ash contents of the APPLS produced by cultures at each pH. The effects of pH on the degradation of a spruce (Picea pungens) [C-lignin]lignocellulose and a Douglas fir (Pseudotsuga menziesii) [C glucan]-lignocellulose were also determined at pH values between 6.5 and 9.5 (0.5 pH unit intervals). The incubations were carried out for 3 weeks at 37 degrees C with bubbler-tube cultures. The percentage of initial C recovered as CO(2), C labeled water-soluble products, and [C]APPL was then determined. The mineralization of lignin and cellulose to CO(2) was optimal at pHs 6.5 and 7.0, respectively. However, the optimum for lignin and cellulose solubilization was pH 8.5, which correlated with the pH 8.5 optimum for APPL production. Overall, the data show that, whereas lignin mineralization is optimal at neutral to slightly acidic pHs, lignocellulose degradation with lignin solubilization and APPL production is promoted by alkaline pHs. These findings indicate that lignin solubilizing actinomycetes may play an important role in the metabolism of lignin in neutral to alkaline soils in which ligninolytic fungi are not highly competitive. PMID- 16347125 TI - Role of Veratryl Alcohol in Regulating Ligninase Activity in Phanerochaete chrysosporium. AB - Ligninase activity in Phanerochaete chrysosporium is stimulated by incubating cultures with various substrates for the enzyme, including veratryl (3,4 dimethoxybenzyl) alcohol, which is a secondary metabolite of this fungus. This study was designed to provide insight into the mechanism involved in this stimulation. Ligninase activity increased 2 to 4 h after the addition of exogenous veratryl alcohol to ligninolytic cultures. This increase was prevented by inhibitors of protein synthesis. Analysis of the extracellular proteins by high-performance anion-exchange liquid chromatography revealed increases in the amounts of some, but not all, ligninase species. The normal rapid decrease in ligninase activity in aging cultures was not prevented or retarded by veratryl alcohol, indicating that veratryl alcohol does not increase ligninase activity by protecting extant enzyme. We conclude that veratryl alcohol probably functions via an induction type of mechanism, affecting only certain ligninase species. Results with an isolated lignin indicate that lignin (or its biodegradation products) functions in the same way that veratryl alcohol does. PMID- 16347126 TI - Utilization of Methanol plus Hydrogen by Methanosarcina barkeri for Methanogenesis and Growth. AB - Methanosarcina barkeri grew on methanol plus H(2). Both substrates were consumed in equimolar amounts. Growth was strictly dependent on the presence of acetate, which was required for the biosynthesis of cellular constituents. Only about 0.4% of the methane produced originated from acetate. By using deuterated methanol, it was demonstrated that methanogenesis from this compound under H(2) did not occur via oxidation of methanol to CO(2) and subsequent reduction but by direct reduction with H(2). Growth yields with methanol plus H(2) and with methanol alone were not significantly different: 2.8 g of cells per mol of methanol in mineral medium and 4.6 g of cells per mol of methanol in complex medium, respectively. Growth of M. barkeri on methanol plus H(2) depended strictly on the presence of sodium ions in the medium. In the presence of 50 mM K the K(s) for Na was 5 mM. PMID- 16347128 TI - Production of Deepoxy-Diacetoxyscirpenol in a Culture of Fusarium graminearum. AB - Deepoxy-diacetoxyscirpenol was isolated from a laboratory culture of Fusarium graminearum grown on a solid rice substrate. It was characterized as 3-hydroxy 4,15-diacetoxy-trichothec-9,12-diene by proton nuclear magnetic resonance and mass spectrometry. This is the first report of the occurrence of this metabolite in a fungus culture. PMID- 16347127 TI - Influence of forage phenolics on ruminal fibrolytic bacteria and in vitro fiber degradation. AB - In vitro cultures of ruminal microorganisms were used to determine the effect of cinnamic acid and vanillin on the digestibility of cellulose and xylan. Cinnamic acid and vanillin depressed in vitro dry matter disappearance of cellulose 14 and 49%, respectively, when rumen fluid was the inoculum. The number of viable Bacteroides succinogenes cells, the predominant cellulolytic organism, was threefold higher for fermentations which contained vanillin than for control fermentations. When xylan replaced cellulose as the substrate, a 14% decrease in the digestibility of xylan was observed with vanillin added; however, the number of viable xylanolytic bacteria cultured from the batch fermentation was 10-fold greater than that of control fermentations. The doubling time of B. succinogenes was increased from 2.32 to 2.58 h when vanillin was added to cellobiose medium, and absorbance was one-half that of controls after 18 h. The growth rate of Ruminococcus albus and Ruminococcus flavefaciens was inhibited more by p-coumaric acid than by vanillin, although no reduction of final absorbance was observed in their growth cycles. Vanillin, and to a lesser extent cinnamic acid, appeared to prevent the attachment of B. succinogenes cells to cellulose particles, but did not affect dissociation of cells from the particles. B. succinogenes, R. albus, R. flavefaciens, and Butyrivibrio fibrisolvens all modified the parent monomers cinnamic acid, p-coumaric acid, ferulic acid, and vanillin, with B. fibrisolvens causing the most extensive modification. These results suggest that phenolic monomers can inhibit digestibility of cellulose and xylan, possibly by influencing attachment of the fibrolytic microorganisms to fiber particles. The reduced bacterial attachment to structural carbohydrates in the presence of vanillin may generate more free-floating fibrolytic organisms, thus giving a deceptively higher viable count. PMID- 16347129 TI - Novel method of lactic Acid production by electrodialysis fermentation. AB - In lactic acid fermentation by Lactobacillus delbrueckii, the produced lactic acid affected the lactic acid productivity. Therefore, for the purpose of alleviating this inhibitory effect, an electrodialysis fermentation method which can continuously remove produced lactic acid from the fermentation broth was applied to this fermentation process. As a result, the continuation of fermentation activity was obtained, and the productivity was three times higher than in non-pH-controlled fermentation. In electrodialysis fermentation, the amount of produced lactic acid was 82.2 g/liter, which was about 5.5 times greater than that produced in non-pH-controlled fermentation. It was concluded that these good results were obtained on account of alleviating the lactic acid inhibitory effect by electrodialysis fermentation. However, the fouling of anion exchange membranes by cells was observed in electrodialysis fermentation. PMID- 16347130 TI - Utilization of Xylan by Yeasts and Its Conversion to Ethanol by Pichia stipitis Strains. AB - Yeasts able to grow on d-xylose were screened for the ability to hydrolyze xylan. Xylanase activity was found to be rare; a total of only 19 of more than 250 strains yielded a positive test result. The activity was localized largely in the genus Cryptococcus and in Pichia stipitis and its anamorph Candida shehatae. The ability to hydrolyze xylan was generally uncoupled from that to hydrolyze cellulose; only three of the xylan-positive strains also yielded a positive test for cellulolytic activity. Of the 19 xylanolytic strains, 2, P. stipitis CBS 5773 and CBS 5775, converted xylan into ethanol, with about 60% of a theoretical yield computed on the basis of the amount of d-xylose present originally that could be released by acid hydrolysis. PMID- 16347131 TI - Occurrence of trichothecin in wheat. AB - Trichothecin, a toxic metabolite of Trichothecium roseum, was detected in 3 of 13 wheat samples examined at levels of 560, 290, and 270 ng/g. In addition, all the T. roseum isolates from the wheat produced the toxin on wheat. PMID- 16347132 TI - Nitrogen Limitation in Natural Populations of Cyanobacteria (Spirulina and Oscillatoria spp.) and Its Effect on Macromolecular Synthesis. AB - Natural populations of the cyanobacteria Spirulina species and Oscillatoria species obtained from Israeli fishponds were limited in growth by nitrogen availability in summer. Physiological indicators for nitrogen limitation, such as phycocyanin, chlorophyll a, and carbohydrate content, did not show clear evidence for nitrogen limited growth, since these organisms are capable of vertical migration from and to the nitrogen-rich bottom. By means of C labeling of the cells under simulated pond conditions followed by cell fractionation into macromolecular compounds, we found that carbohydrates synthesized at the lighted surface were partially utilized for dark protein synthesis at the bottom of these ponds. PMID- 16347133 TI - Kinetics of phenol biodegradation by an immobilized methanogenic consortium. AB - A phenol-degrading methanogenic enrichment was successfully immobilized in agar as shown by the stoichiometric conversion of phenol to CH(4) and CO(2). The enrichment contained members of three physiological groups necessary for the syntrophic mineralization of phenol: a phenol-oxidizing bacterium, a Methanothrix like bacterium, and an H(2)-utilizing methanogen. The immobilization technique resulted in the cells being embedded in a long, thin agar strand (1 mm in diameter by 2 to 50 cm in length) that resembled spaghetti. Immobilization had three effects as shown by a comparative kinetic analysis of phenol degradation by free versus immobilized cells. (i) The maximum rate of degradation was reduced from 14.8 to 10.0 mug of phenol per h; (ii) the apparent K(m) for the overall reaction was reduced from 90 to 46 mug of phenol per ml, probably because of the retention of acetate, H(2) and CO(2) in the proximity of immobilized methanogens; and (iii) the cells were protected from substrate inhibition caused by high concentrations of phenol, which increased the apparent K(i) value from 900 to 1,725 mug of phenol per ml. Estimates for the kinetic parameters K(m), K(i), and V(max) were used in a modified substrate inhibition model that simulated rates of phenol degradation for given phenol concentrations. The simulated rates were in close agreement with experimentally derived rates for both stimulatory and inhibitory concentrations of phenol. PMID- 16347134 TI - Utilization of Lactate Isomers by Propionibacterium freudenreichii subsp. shermanii: Regulatory Role for Intracellular Pyruvate. AB - Five strains of Propionibacterium freudenreichii subsp. shermanii utilized the l (+) isomer of lactate at a faster rate than they did the d-(-) isomer when grown with a mixture of lactate isomers under a variety of conditions. ATCC 9614, grown anaerobically in defined medium containing 160 mM dl-lactate, utilized only 4 and 15% of the d-(-)-lactate by the time 50 and 90%, respectively, of the l-(+) lactate was used. The intracellular pyruvate concentration was high (>100 mM) in the initial stages of lactate utilization, when either dl-lactate or the l-(+) isomer was the starting substrate. The concentration of this intermediate dropped during dl-lactate fermentation such that when only d-(-)-lactate remained, the concentration was <20 mM. When only the d-(-) isomer was initially present, a similar relatively low concentration of intracellular pyruvate was present, even at the start of lactate utilization. The NAD-independent lactate dehydrogenase activities in extracts showed different kinetic properties with regard to pyruvate inhibition, depending upon the lactate isomer present. Pyruvate gave a competitive inhibitor pattern with l-(+)-lactate and a mixed-type inhibitor pattern with d-(-)-lactate. It is suggested that these properties of the lactate dehydrogenases and the intracellular pyruvate concentrations explain the preferential use of the l-(+) isomer. PMID- 16347135 TI - Metabolism of Aspartate by Propionibacterium freudenreichii subsp. shermanii: Effect on Lactate Fermentation. AB - More than 90% of the aspartate in a defined medium was metabolized after lactate exhaustion such that 3 mol of aspartate and 1 mol of propionate were converted to 3 mol of succinate, 3 mol of ammonia, 1 mol of acetate, and 1 mol of CO(2). This pathway was also evident when propionate and aspartate were the substrates in complex medium in the absence of lactate. In complex medium with lactate present, about 70% of the aspartate was metabolized to succinate and ammonia during lactate fermentation, and as a consequence of aspartate metabolism, more lactate was fermented to acetate and CO(2) than was fermented to propionate. The conversion of aspartate to fumarate and ammonia by the enzyme aspartase and subsequent reduction of fumarate to succinate occurred in the five strains of Propionibacterium freudenreichii subsp. shermanii studied. The ability to metabolize aspartate in the presence of lactate appeared to be related to aspartase activity. The specific activity of aspartase increased during and after lactate utilization, and the levels of this enzyme were lower in cells grown in defined medium than levels in those cells grown in complex medium. Under the conditions used, no other amino acids were readily metabolized in the presence of lactate. The possibility that aspartate metabolism by propionibacteria in Swiss cheese has an influence on CO(2) production is discussed. PMID- 16347136 TI - Transfer of Plasmids to an Antibiotic-Sensitive Mutant of Zymomonas mobilis. AB - Wild-type strains of Zymomonas mobilis exhibit multiple antibiotic resistance and thus restrict the use of many broad-host-range plasmids in them as cloning vehicles. Antibiotic-sensitive mutants of Z. mobilis were isolated and used as hosts for the conjugal transfer of broad-host-range plasmids from Escherichia coli. Such antibiotic-sensitive strains can facilitate the application of broad host-range plasmids to the study of Z. mobilis. PMID- 16347137 TI - Quantitation of adsorption of rhizobia in low numbers to small legume roots. AB - Bacteria adsorbed in low numbers to alfalfa or clover root surfaces were counted after incubation of seedlings in mineral solution with very dilute inocula (less than 10 bacteria per ml) of an antibiotic-resistant strain under defined conditions. After specified washing, bacteria which remained adsorbed to roots were selectively quantitated by culturing the roots embedded in yeast extract mannitol-antibiotic agar and counting the microcolonies along the root surface; the range was from about 1 bacterium per root (estimated as the most probable number) to 50 bacteria per cm of root length (by direct counting). This simple procedure can be used with any pair of small-rooted plant and antibiotic resistant bacterium, requires bacterial concentrations comparable to those frequently found in soils, and yields macroscopic localization and distribution data for adsorbed bacteria over the root surface. The number of adsorbed bacteria was proportional to the size of the inoculum. One of every four Rhizobium meliloti cells adsorbed in very low numbers to alfalfa roots resulted in the formation of a nodule. Overall adsorption of various symbiotic and nonsymbiotic bacterial strains to alfalfa and clover roots did not reflect the specificities of these legumes for their respective microsymbionts, R. meliloti and R. trifolii. PMID- 16347139 TI - Aerobic metabolism of trichloroethylene by a bacterial isolate. AB - A number of soil and water samples were screened for the biological capacity to metabolize trichloroethylene. One water sample was found to contain this capacity, and a gram-negative, rod-shaped bacterium which appeared to be responsible for the metabolic activity was isolated from this sample. The isolate degraded trichloroethylene to CO(2) and unidentified, nonvolatile products. Oxygen and water from the original site of isolation were required for degradation. PMID- 16347138 TI - Host-Symbiont Specificity Expressed during Early Adsorption of Rhizobium meliloti to the Root Surface of Alfalfa. AB - Early (4 h) adsorption of Rhizobium meliloti L5-30 in low numbers to alfalfa roots in mineral solution was examined for competition with other bacterial strains. All tested competitor strains decreased the adsorption of L5-30 by extents which depended on the strain and its concentration. The decrease of adsorption by R. meliloti competitors (all of them infective in alfalfa) was nearly complete at saturation (97 to 99% decrease). All other heterologous rhizobia and Agrobacterium tumefaciens at saturating concentrations (10 to 10 per ml) decreased adsorption of L5-30 only partially, less than 60%. The differential effects of homologous and heterologous competitors indicate that initial adsorption of R. meliloti to the root surface of its host occurs in symbiont specific as well as nonspecific modes and suggest the existence of binding sites on roots which are highly selective for the specific microsymbiont in the presence of other heterologous bacteria even in very unfavorable (less than 10) symbiont-competitor concentration ratios. PMID- 16347140 TI - Comparison of yeast growth in mesquite wood hydrolysate. AB - Hot-water extracts of mesquite (Prosopis glandulosa) wood were assayed for their total carbohydrate, reducing sugar, and glucose content. These hydrolysates were then used as complete media for yeast growth. A total of 10 strains of yeasts were evaluated for their biomass production in the mesquite wood hydrolysates. Levels of utilizable carbohydrate proved to be the limiting factor for yeast growth in the hydrolysates. PMID- 16347141 TI - Long-term storage of plant-pathogenic bacteria in sterile distilled water. AB - This study was made to determine the effectiveness of the preservation of plant pathogenic bacteria in sterile distilled water. After 20 or 24 years of storage in distilled water, a very high percentage (90 to 92%) of the isolates of Agrobacterium tumefaciens and Pseudomonas spp. were still alive. Moreover, 12 of 13 viable (after 24 years) isolates of P. syringae subsp. syringae maintained their ability to produce syringomycin and were pathogenic to bean seedlings. The water-stored cells of two isolates of P. syringae subsp. syringae, when observed by electron microscopy, were smaller than cells of 24-h-old subcultures of bacterial cells grown in nutrient broth; the water-stored cells appeared plasmolysed with an electron-dense cytoplasm and thickened cell wall. PMID- 16347142 TI - Ability of pH-Selected Mixed Ruminal Microbial Populations to Digest Fiber at Various pHs. AB - Inocula from pH 5.5-selected rumen microorganisms failed to digest fiber any better in batch culture at pH 5.0 or 5.5 than did inocula from pH 6.5-selected organisms. Furthermore, upon extended batch incubation, both the viable cellulolytic and amylolytic bacteria became increasingly associated with the fiber portion and less with the liquid portion of the cultures. PMID- 16347143 TI - Cloning, Expression, and Purification of Glutamine Synthetase from Clostridium acetobutylicum. AB - A glutamine synthetase (GS) gene, glnA, from the gram-positive obligate anaerobe Clostridium acetobutylicum was cloned on recombinant plasmid pHZ200 and enabled Escherichia coli glnA deletion mutants to utilize (NH(4))(2)SO(4) as a sole source of nitrogen. The cloned C. acetobutylicum gene was expressed from a regulatory region contained within the cloned DNA fragment. glnA expression was subject to nitrogen regulation in E. coli. This cloned glnA DNA did not enable an E. coli glnA ntrB ntrC deletion mutant to utilize arginine or low levels of glutamine as sole nitrogen sources, and failed to activate histidase activity in this strain which contained the Klebsiella aerogenes hut operon. The GS produced by pHZ200 was purified and had an apparent subunit molecular weight of approximately 59,000. There was no DNA or protein homology between the cloned C. acetobutylicum glnA gene and GS and the corresponding gene and GS from E. coli. The C. acetobutylicum GS was inhibited by Mg in the gamma-glutamyl transferase assay, but there was no evidence that the GS was adenylylated. PMID- 16347144 TI - Identification of eubacteria isolated from leaf cavities of four species of the N fixing azolla fern as arthrobacter conn and dimmick. AB - The nutritional and physiological characteristics of 15 isolates from four species of the Azolla fern were determined. Although some minor variation existed in levels of urease activity, ability to utilize xylose, and formation of N(2) gas from NO(3), all 15 isolates were rather similar and believed to represent a single species. These eubacteria exhibited aminopeptidase activity and became viscous when treated with KOH, similar to gram-negative organisms; however, the absence of lipopolysaccharide and 2-keto-3-deoxyoctonate in cell walls indicated that they are truly gram-positive organisms. They are unusual because peptidoglycan could not be detected during most of their growth cycle. The presence of lysine as the major diamino acid in cell wall hydrolysates, the inability to hydrolyze cellulose, and the distinctive developmental pattern with rods and "V" forms present during log phase, becoming progressively shorter until cocci dominated during stationary and death phases, indicated that these organisms belong to the genus Arthrobacter Conn and Dimmick. With the exception of the inability to hydrolyze gelatin, their characteristics are consistent with those of the type species, Arthrobacter globiformis Conn and Dimmick. PMID- 16347145 TI - Variability of the influence of physicochemical factors affecting bacterial adhesion to polystyrene substrata. AB - The role of electrostatic and hydrophobic interactions and solid and liquid surface tensions in the adhesion of four bacterial species (Pseudomonas fluorescens, Enterobacter cloacae, Chromobacterium sp., and Flexibacter sp.) to hydrophobic polystyrene petri dishes and to more hydrophilic polystyrene tissue culture dishes was investigated. The effect of electrostatic interactions was investigated by determining the effects of different electrolyte solutions on attachment to and of different electrolyte and pH solutions on detachment from the polystyrene substrate. The significance of solid and liquid surface tensions and hydrophobic interactions was investigated by measuring the effects of different surfactants (including a concentration series of dimethyl sulfoxide) on adhesion and detachment. Adhesion varied with bacterial species, substratum, and electrolyte type and concentration, with no apparent correlation between adhesion and electrolyte valence or concentration. The influence of different pH and detergent solutions on bacterial detachment also varied with species, substratum, pH, and detergent type; however, the greatest degree of detachment of all strains from the surfaces was produced by detergent treatment. The results suggest that adhesion cannot be attributed to any one type of adhesive interaction. There was some evidence for both electrostatic and hydrophobic interactions, but neither interaction could wholly account for the data. PMID- 16347146 TI - Alginate production by plant-pathogenic pseudomonads. AB - Eighteen plant-pathogenic and three non-plant-pathogenic pseudomonads were tested for the ability to produce alginic acid as an exopolysaccharide in vitro. Alginate production was demonstrated for 10 of 13 fluorescent plant-pathogenic pseudomonads tested with glucose or gluconate as the carbon source, but not for all 5 nonfluorescent plant pathogens and all 3 non-plant pathogens tested. With sucrose as the carbon source, some strains produced alginate while others produced both polyfructan (levan) and alginate. Alginates ranged from <1 to 28% guluronic acid, were acetylated, and had number-average molecular weights of 11.3 x 10 to 47.1 x 10. Polyfructans and alginates were not elicitors of the soybean phytoalexin glyceollin when applied to wounded cotyledon surfaces and did not induce prolonged water soaking of soybean leaf tissues. All or most pseudomonads in rRNA-DNA homology group I may be capable of synthesizing alginate as an exopolysaccharide. PMID- 16347147 TI - Decomposition Studies in Two Central Ontario Lakes Having Surficial pHs of 4.6 and 6.6. AB - The rates of cellulose breakdown, composition of detrital microflora, and density of bacterial populations were determined in the epilimnetic sediments and water columns of two poorly buffered, oligotrophic, Canadian Shield lakes having mean surficial pHs of 4.6 (Bat Lake) and 6.6 (Harp Lake). The decomposition rate was significantly lower in oxic sediment of the acidified lake than of the circumneutral lake, but water column rates were almost identical in the two lakes. These results are explained in terms of the groups of cellulolytic microorganisms which were observed by phase-contrast microscopy as being active at the different sites: fungi in Bat Lake water and Cytophaga-like bacteria in the water and sediment of Harp Lake. Cytophaga-like bacteria were also the main decomposers in Bat Lake sediment, but their activity was restricted at porewater pHs of <5.0. Acridine orange direct counts of bacteria in the top centimeter of sediment ranged from 3.7 x 10 to 1.0 x 10 per g, and counts in planktonic water samples ranged from 4.9 x 10 to 1.2 x 10 per ml. Bacterial densities at most sites decreased significantly (P < 0.001) from August to late October, but did not show a consistent pattern of differences related to pH. PMID- 16347148 TI - Degradation of dead microbial biomass in a marine sediment. AB - The availability of dead microbial biomass in a marine beach sand to degradation and mineralization was examined. Microbial sand populations were labeled with [C]glutamic acid, [H]adenine, or [H]thymidine and killed with chloroform. Live sand or seawater (or both) was added to the sterile labeled sand, and biochemical components of the populations were monitored for 10 days. Labeled RNA was degraded more quickly than labeled DNA, but both nucleic acids were degraded to approximately the same extent (60 to 70%). H(2)O was a major acid-soluble breakdown product. RNA (and possibly DNA) breakdown products were reincorporated into DNA (and possibly RNA) during the incubation period. In addition to metabolite salvage, 32% of the total macromolecular C was respired in the 10-day period regardless of whether sand or seawater was used as the inoculum. Respiration was essentially complete in 3 days, whereas nucleic acid degradation continued throughout the 10-day incubation. The results indicate that dead microbial biomass is a labile component of the sediment ecosystem. PMID- 16347149 TI - Close association of azospirillum and diazotrophic rods with different root zones of kallar grass. AB - The populations of diazotrophic and nondiazotrophic bacteria were estimated in the endorhizosphere and on the rhizoplane of Kallar grass (Leptochloa fusca) and in nonrhizosphere soil. Microaerophilic diazotrophs were counted by the most probable-number method, using two semisolid malate media, one of them adapted to the saline-sodic Kallar grass soil. Plate counts of aerobic heterotrophic bacteria were done on nutrient agar. The dominating N(2)-fixing bacteria were differentiated by morphological, serological, and physiological criteria. Isolates, which could not be assigned to a known species, were shown to fix nitrogen unequivocally by N(2) incorporation. On the rhizoplane we found 2.0 x 10 diazotrophs per g (dry weight) of root, which consisted in equal numbers of Azospirillum lipoferum and Azospirillum-like bacteria showing characteristics different from those of known Azospirillum species. Surface sterilization by NaOCI treatment effectively reduced the rhizoplane population, so that bacteria released by homogenization of roots could be regarded as endorhizosphere bacteria. Azospirillum spp. were not detected in the endorhizosphere, but diazotrophic, motile, straight rods producing a yellow pigment occurred with 7.3 x 10 cells per g (dry weight) of root in the root interior. In nonrhizosphere soil we found 3.1 x 10 nitrogen-fixing bacteria per g. Diazotrophs were preferentially enriched in the Kallar grass rhizosphere. In nonrhizosphere soil they made up 0.2% of the total aerobic heterotrophic microflora, on the rhizoplane they made up 7.1%, and in the endorhizosphere they made up 85%. Owing to high numbers in and on roots and their preferential enrichment, we concluded that diazotrophs are in close association with Kallar grass. They formed entirely different populations on the rhizoplane and in the endorhizosphere. PMID- 16347150 TI - Antagonism of Lactic Acid Bacteria against Phytopathogenic Bacteria. AB - A variety of lactic acid bacteria, isolated from plant surfaces and plant associated products, were found to be antagonistic to test strains of the phytopathogens Xanthomonas campestris, Erwinia carotovora, and Pseudomonas syringae. Effective "in vitro" inhibition was found both on agar plates and in broth cultures. In pot trials, treatment of bean plants with a Lactobacillus plantarum strain before inoculation with P. syringae caused a significant reduction of the disease incidence. PMID- 16347151 TI - Magnesium and manganese content of halophilic bacteria. AB - Magnesium and manganese contents were measured by atomic absorption spectrophotometry in bacteria of several halophilic levels, in Vibrio costicola, a moderately halophilic eubacterium growing in 1 M NaCl, Halobacterium volcanii, a halophilic archaebacterium growing in 2.5 M NaCl, Halobacterium cutirubrum, an extremely halophilic archaebacterium growing in 4 M NaCl, and Escherichia coli, a nonhalophilic eubacterium growing in 0.17 M NaCl. Magnesium and manganese contents varied with the growth phase, being maximal at the early log phase. Magnesium and manganese molalities in cell water were shown to increase with the halophilic character of the logarithmically growing bacteria, from 30 mmol of Mg per kg of cell water and 0.37 mmol of Mn per kg of cell water for E. coli to 102 mmol of Mg per kg of cell water and 1.6 mmol of Mn per kg of cell water for H. cutirubrum. The intracellular concentrations of manganese were determined independently by a radioactive tracer technique in V. costicola and H. volcanii. The values obtained by Mn loading represented about 70% of the values obtained by atomic absorption. The increase of magnesium and manganese contents associated with the halophilic character of the bacteria suggests that manganese and magnesium play a role in haloadaptation. PMID- 16347152 TI - New procedure for extraction of ammonium from natural waters for N isotopic ratio determinations. AB - A new method for extracting ammonium from natural waters for N isotopic ratio determination is described. The method employs the conversion of the ammonium nitrogen into indophenol, which is then concentrated onto an octadecylsilane column. The method shows accuracy and precision comparable to those of other methods described in the literature. Some results from field experiments on the Swedish west coast are presented. PMID- 16347153 TI - Differential Effects of Lithium Chloride on In Vitro Growth of Clavibacter michiganense subsp. nebraskense Depending upon Inoculum Source. AB - The bacterium Clavibacter michiganense subsp. nebraskense (Corynebacterium michiganense subsp. nebraskense) was grown in broth cultures and inoculated into corn plants. The plating efficiency of cells from broth cultures was essentially the same on nutrient broth-yeast extract and the semiselective medium for this bacterium, CNS. However, when cells were isolated from Goss bacterial wilt- and blight-infected corn, very few were recovered on CNS compared with the amount recovered on nutrient broth-yeast extract agar. When lithium chloride was omitted from the CNS, recoveries from infected corn were nearly the same as on nutrient broth-yeast extract agar. No other ingredient of CNS was inhibitory, nor did substitution of other salts for lithium chloride cause equal inhibition. The amount of inhibition was proportional to lithium chloride concentration. The inhibition by lithium chloride occurred with several strains of the bacterium isolated from one corn cultivar and with one of the strains recovered from three different cultivars of infected corn. PMID- 16347154 TI - Direct acridine orange counting of bacteria preserved with acidified lugol iodine. AB - Acidified and nonacidified Lugol iodine solution was tested under several storage temperatures and at several times as a preservative for marine bacteria. Direct counts with acridine orange showed no significant difference between glutaraldehyde- and Lugol iodine solution-preserved samples under any storage temperature when samples were counted within 1 week of collection. Specimens in long-term (up to 6 months) storage required refrigeration and treatment with acidified Lugol iodine solution for adequate preservation. Lugol iodine solution preserved bacteria appeared intact under scanning electron microscopy. Lugol iodine solution did not preserve chlorophyll autofluorescence in phytoplankton. PMID- 16347155 TI - Biodelignification of lemon grass and citronella bagasse by white-rot fungi. AB - Twelve white-rot fungi were grown in solid-state culture on lemon grass (Cymbopogon citratus) and citronella (Cymbopogon winterianus) bagasse. The two lignocellulosic substrates had 11% permanganate lignin and a holocellulose fraction of 58%. After 5 to 6 weeks at 20 degrees C, nine fungi produced a solid residue from lemon grass with a higher in vitro dry matter enzyme digestibility than the original bagasse; seven did the same for citronella. The best fungus for both substrates was Bondarzewia berkeleyi; it increased the in vitro dry matter enzyme digestibility to 22 and 24% for lemon grass and citronella, respectively. The increases were correlated with weight loss and lignin loss. All fungi decreased lignin contents: 36% of the original value for lemon grass and 28% for citronella. Practically all fungi showed a preference for hemicellulose over cellulose. PMID- 16347156 TI - Effect of Sulfur-Containing Compounds on Growth of Methanosarcina barkeri in Defined Medium. AB - Methanosarcina barkeri Fusaro (DSM 804) could grow on methanol in a mineral medium containing cysteine or thiosulfate as the sole sulfur source. Optimum growth occurred at cysteine concentrations of 1 to 2.8 mM and at thiosulfate concentrations of 2.5 to 5 mM. No inhibition of growth was observed even when these concentrations were doubled in the culture medium. Under the optimum cysteine and thiosulfate concentrations, the generation times of the organism were about 8 to 10 and 10 to 12 h, respectively, giving a cell yield of about 0.14 to 0.17 and 0.08 to 0.11 g (dry weight)/g of methanol consumed. The organism metabolized cysteine and thiosulfate during growth, giving rise to sulfide in the culture medium. H(2)S evolution from cysteine and thiosulfate was catalyzed by two enzymes, namely cysteine desulfhydrase and thiosulfate reductase, respectively, as revealed by enzyme assay in the crude cell-free extract of the organism. PMID- 16347157 TI - Acetate Synthesis from H(2) plus CO(2) by Termite Gut Microbes. AB - Gut microbiota from Reticulitermes flavipes termites catalyzed an H(2)-dependent total synthesis of acetate from CO(2). Rates of H(2)-CO(2) acetogenesis in vitro were 1.11 +/- 0.37 mumol of acetate g (fresh weight) h (equivalent to 4.44 +/- 1.47 nmol termite h) and could account for approximately 1/3 of all the acetate produced during the hindgut fermentation. Formate was also produced from H(2) + CO(2), as were small amounts of propionate, butyrate, and lactate-succinate. However, H(2)-CO(2) formicogenesis seemed largely unrelated to acetogenesis and was believed not to be a significant reaction in situ. Little or no CH(4) was formed from H(2) + CO(2) or from acetate. H(2)-CO(2) acetogenesis was inhibited by O(2), KCN, CHCl(3), and iodopropane and could be abolished by prefeeding R. flavipes with antibacterial drugs. By contrast, prefeeding R. flavipes with starch resulted in almost complete defaunation but had little effect on H(2) CO(2) acetogenesis, suggesting that bacteria were the acetogenic agents in the gut. H(2)-CO(2) acetogenesis was also observed with gut microbiota from Prorhinotermes simplex, Zootermopsis angusticollis, Nasutitermes costalis, and N. nigriceps; from the wood-eating cockroach Cryptocercus punctulatus; and from the American cockroach Periplaneta americana. Pure cultures of H(2)-CO(2)-acetogenic bacteria were isolated from N. nigriceps, and a preliminary account of their morphological and physiological properties is presented. Results indicate that in termites, CO(2) reduction to acetate, rather than to CH(4), represents the main electron sink reaction of the hindgut fermentation and can provide the insects with a significant fraction (ca. 1/3) of their principal oxidizable energy source, acetate. PMID- 16347158 TI - Preliminary characterization of bacteriophages infecting the thermophilic actinomycete thermomonospora. AB - Bacteriophages lysing strains of Thermomonospora alba and T. fusca were isolated, following specific enrichment, from vegetable composts. Four Thermomonospora phages were distinguished by plaque morphology and host range. Electron microscopy of phage particles, termperature inactivation profiles, and electrophoretic analyses of major virion proteins and genomic DNA were used in the comparison and initial characterization of these phages. The four phages studied possessed polyhedral heads and long tails; genomes were linear double stranded DNA molecules, 35 to 45 kilobases in length, which probably contain cohesive ends. Transfection of Thermomonospora protoplasts with purified genomic DNA from one of the phages was demonstrated. PMID- 16347159 TI - Production of l-Phenylalanine from Starch by Analog-Resistant Mutants of Bacillus polymyxa. AB - p-Fluorophenylalanine-resistant mutants of starch-degrading Bacillus polymyxa ATCC 842, generated by ethyl methanesulfonate mutagenesis followed by incubation with caffeine, overproduced small amounts of l-phenylalanine (l-phe) from starch. A beta-2-thienylalanine-resistant mutant (BT-7) derived from p fluorophenylalanine mutant (C-4000 FP-4) and resistant to both p fluorophenylalanine and beta-2-thienylalanine produced 0.5 g of l-phe and 0.15 g of l-tyrosine per liter from 10 g of starch per liter when growing in a minimal medium. trans-Cinnamic acid (CA) was also excreted by both mutants, indicating the possibility of l-phenylalanine ammonia-lyase-induced deamination of l-phe to CA. The amount of l-phe-derived CA detected in BT-7 was less compared with mutant C-4000 FP-4. CA production was induced in the parent only when l-phe was used as a sole nitrogen source. Time of CA production in the two mutants could be delayed by addition of other nitrogen sources, an indication of possible l-phenylalanine ammonia-lyase inhibition or repression. The presence of l-phenylalanine ammonia lyase in B. polymyxa mutant C-4000 FP-4 was confirmed by assays of cell-free extracts from cells grown in starch minimal medium containing l-phe as the sole nitrogen source. Preliminary studies of the regulation of deoxy-d-arabino heptulosonate-7-phosphate synthase and prephenate dehydratase in the wild-type strain showed that deoxy-d-arabino-heptulosonate-7-phosphate synthase was subject to feedback inhibition by l-phe, l-tyrosine, and l-tryptophan. Inhibition by each amino acid was to a similar extent singly or in combination at a 0.5 mM level of each amino acid. Prephenate dehydratase was feedback inhibited by l-phe, but not by l-tyrosine or l-tryptophan or both. In the double analog-resistant mutant BT 7, deoxy-d-arabino-heptulosonate-7-phosphate synthase had specific activity similar to that in the wild type, and the enzyme was still subject to feedback inhibition. However, prephenate dehydratase had increased specific activity and it was also insensitive to feedback inhibition by l-phe. The overproduction of aromatic amino acids by BT-7 was thought to be due, at least in part, to deregulation of feedback inhibition of prephenate dehydratase. Chorismate mutase was not subject to feedback inhibition in the wild type and was unaffected in the mutant. PMID- 16347160 TI - Simplified method for dissolved DNA determination in aquatic environments. AB - A method was developed for the determination of dissolved DNA in aquatic environments. The method is based upon the concentration of dissolved DNA by ethanol precipitation of 0.2-mum-pore-size filtered water. The DNA in concentrated extracts was quantified by the fluorescence of Hoechst 33258-DNA complexes. Fluorescence not attributable to DNA was corrected for by DNase I digestion of the extracts and averaged 25% of the total fluorescence for all samples. The effectiveness of the procedure for concentrating dissolved DNA was demonstrated by the efficient (>90%) recovery of internal standards. Concentrations of dissolved DNA from a variety of marine and freshwater environments ranged from 0.2 to 44 mug/liter, with the highest values being obtained for estuarine and river environments. The method is simple, specific for DNA, and more sensitive than previously described methods for the determination of extracellular DNA. PMID- 16347161 TI - Affinity Purifications of Aldose Reductase and Xylitol Dehydrogenase from the Xylose-Fermenting Yeast Pachysolen tannophilus. AB - Although xylose is a major product of hydrolysis of lignocellulosic materials, few yeasts are able to convert it to ethanol. In Pachysolen tannophilus, one of the few xylose-fermenting yeasts found, aldose reductase and xylitol dehydrogenase were found to be key enzymes in the metabolic pathway for xylose fermentation. This paper presents a method for the rapid and simultaneous purification of both aldose reductase and xylitol dehydrogenase from P. tannophilus. Preliminary studies indicate that this method may be easily adapted to purify similar enzymes from other xylose-fermenting yeasts. PMID- 16347162 TI - Comparative study of the ability of three xanthobacter species to metabolize cycloalkanes. AB - The ability of three species of Xanthobacter to metabolize cyclohexane and its derivatives has been compared. Xanthobacter flavus was unable to utilize any of the cycloalkanes under investigation. X. autotrophicus was unable to utilize cyclohexane but was able to grow with a limited range of substituted cycloalkanes, including cyclohexanol and cyclohexanone. Comparison of a previously isolated cyclohexane growing Xanthobacter sp. with X. flavus and X. autotrophicus indicated it to be closely related to X. autotrophicus. Studies with cell-free extracts have indicated that the route of metabolism for cyclohexanol by X. autotrophicus is the same as that shown for the cyclohexane growing Xanthobacter sp., proceeding via cyclohexanol-->cyclohexanone--> epsilon caprolactone-->--> adipic acid. A comparison of the cyclohexanol dehydrogenase found in X. autotrophicus with that found in the cyclohexane-growing Xanthobacter sp. indicated these enzymes to be distinctly different from one another on the basis of substrate specificity, molecular weight, and pH optima. The cyclohexanone monooxygenase enzymes found in the two bacteria were also found to be different when the pH optima and cofactor specificity of the two enzymes were compared. Preliminary genetic studies on the cyclohexane-growing Xanthobacter sp. have indicated that there are no plasmids present in this bacterium. The presence of RP4 in the Xanthobacter sp. can be detected following its conjugation with an RP4-carrying Escherichia coli strain. PMID- 16347163 TI - Measurement of Glucose Utilization by Pseudomonas fluorescens That Are Free Living and That Are Attached to Surfaces. AB - The assimilation and respiration of glucose by attached and free-living Pseudomonas fluorescens were compared. The attachment surfaces were polyvinylidene fluoride, polyethylene, and glass. Specific uptake of [C]glucose was determined after bacterial biomass was measured by (i) microscopic counts or (ii) prelabeling of cells by providing [H]leucine as substrate, followed by dual labeling scintillation counting. The glucose concentration was 1.4, 3.5, 5.5, 7.6, or 9.7 muM. Glucose assimilation by cells which became detached from the surfaces during incubation with glucose was also measured after the detached cells were collected by filtration. The composition of the substratum had no effect on the amount of glucose assimilated by attached cells. Glucose assimilation by attached cells exceeded that by free-living cells by a factor of between 2 and 5 or more, and respiration of glucose by surface-associated cells was greater than that by free-living bacteria. Glucose assimilation by detached cells was greater than that by attached bacteria. Measurements of biomass by microscopic counts gave more consistent results that those obtained with dual labeling, but in general, results obtained by both methods were corroborative. PMID- 16347164 TI - Denitrification by Chromobacterium violaceum. AB - One host (Rana catesbiana)-associated and two free-living mesophilic strains of bacteria with violet pigmentation and biochemical characteristics of Chromobacterium violaceum were isolated from freshwater habitats. Cells of each freshly isolated strain and of strain ATCC 12472 (the neotype strain) grew anaerobically with glucose as the sole carbon and energy source. The major fermentation products of cells grown in Trypticase soy broth (BBL Microbiology Systems, Cockeysville, Md.) supplemented with glucose included acetate, small amounts of propionate, lactate, and pyruvate. The final cell yield and culture growth rate of each strain cultured anaerobically in this medium increased approximately twofold with the addition of 2 mM NaNO(3). Final growth yields increased in direct proportion to the quantity of added NaNO(3) over the range of 0.5 to 5 mM. Each strain reduced NO(3), producing NO(2), NO, and N(2)O. NO(2) accumulated transiently. With 2 mM NaNO(3) in the medium, N(2)O made up 85 to 98% of the N product recovered with each strain. N-oxides were recovered in the same quantity and distribution whether 0.01 atm (ca. 1 kPa) of C(2)H(2) (added to block N(2)O reduction) was present or not. Neither N(2) production nor gas accumulation was detected during NO(3) reduction by growing cells. Cell growth in media containing 0.5 to 5 mM NaNO(2) in lieu of NaNO(3) was delayed, and although N(2)O was produced by the end of growth, NO(2) -containing media did not support growth to an extent greater than did medium lacking NO(3) or NO(2). The data indicate that C. violaceum cells ferment glucose or denitrify, terminating denitrification with the production of N(2)O, and that NO(2) reduction to N(2)O is not coupled to growth but may serve as a detoxification mechanism. No strain detectably fixed N(2) (reduced C(2)H(2)). PMID- 16347165 TI - Purification and Biochemical Analysis of the Cytoplasmic Membrane from the Desiccation-Tolerant Cyanobacterium Nostoc commune UTEX 584. AB - The cytoplasmic membrane of the heterocystous cyanobacterium Nostoc commune UTEX 584 was isolated free of thylakoids and phycobiliprotein-membrane complexes by flotation centrifugation. Purified membranes had a buoyant density of 1.07 g cm and were bright orange. Twelve major proteins were detected in the membrane, and of these, the most abundant had molecular masses of 83, 71, 68, 51, and 46 kilodaltons. The ester-linked fatty acids of the methanol fraction contained 16:0, 18:0, 18:1omega9c, 20:0, and 20:3omega3 with no traces of hydroxy fatty acids. Compound 20:3omega3 represented 56.8% of the total fatty acid methyl esters, a feature which distinguishes the cell membrane of N. commune UTEX 584 from those of all other cyanobacteria which have been characterized to date. Fatty acid 18:3 was not detected. Carotenoids were analyzed by highperformance liquid chromatography. The cytoplasmic membrane contained beta-carotene and echinenone as the dominant carotenoids and lacked chlorophyll a and pheophytin a. Whole cells contained beta-carotene and echinenone, and lesser amounts of zeaxanthin and (3R)-cryptoxanthin. PMID- 16347166 TI - Anaerobic fermentation of woody biomass pretreated with supercritical ammonia. AB - The degradability of ground hardwood by thermophilic anaerobic bacteria (Clostridium thermocellum with or without Thermoanaerobacter strain B6A) was greatly enhanced by pretreatment of the substrate with supercritical ammonia. Relative to C. thermocellum monocultures, cocultures of C. thermocellum and Thermoanaerobacter strain B6A degraded 1.5-fold more pretreated soft maple but produced 2- to 5-fold more fermentation endproducts because Thermoanaerobacter sp. removed reducing sugars produced by C. thermocellum during the fermentation. Dry weight losses were not totally accounted for in end products, due to formation of partially degraded material (<0.4 mum diameter wood particles) during the fermentation. One pretreated hardwood, Southern red oak, was fermented poorly because it released soluble inhibitors at the 60 degrees C incubation temperature. Considerable (6- to 11-fold) increases in substrate degradability were also noted for supercritical ammonia-pretreated wood materials fermented in an in vitro rumen digestibility assay. Degradation of pretreated softwoods by either thermophilic or mesophilic fermentation was not measurable under the conditions tested. PMID- 16347167 TI - Palladium-Mediated Hydrogenation of Unsaturated Hydrocarbons with Hydrogen Gas Released during Anaerobic Cellulose Degradation. AB - Among five hydrogenation catalysts, palladium on charcoal was the most reactive one when suspended in anaerobic culture medium, and Lindlar catalyst (Pd on CaCO(3)) was the most reactive one when suspended in the gas phase of culture tubes. Palladium on charcoal in the culture medium (40 to 200 mg 10 ml) completely inhibited growth of Neocallimastix frontalis and partly inhibited Ruminococcus albus. Lindlar catalyst (40 to 200 mg per tube) suspended in a glass pouch above the culture medium did not affect the rate of cellulose degradation or the ratio of fermentation products by these organisms. Acetylene added to tubes containing Lindlar catalyst in pouches, and either of the two organisms in monoculture or coculture with Methanospirillum hungatei, was reduced to ethylene and then ethane, followed by hydrogen production. Similar results were obtained with 1-pentene. Neither acetylene nor 1-pentene affected cellulose degradation but both inhibited methanogenesis. In the presence of Lindlar catalyst and propylene or 1-butene, fermenter-methanogen cocultures continued to produce methane at the same rate as controls and no olefin reduction occurred. Upon addition of bromoethanesulfonic acid, methanogenesis stopped and olefin reduction took place followed by hydrogen evolution. In a gas mixture consisting of propylene, 1-butene, and 1-pentene, the olefins were reduced at rates which decreased with increasing molecular size. These results demonstrate the technical feasibility of combining in one reactor the volatile fatty acid production by anaerobic digestion with chemical catalyst-mediated reductions, using the valuable by-product hydrogen. PMID- 16347168 TI - Availability of ferric iron for microbial reduction in bottom sediments of the freshwater tidal potomac river. AB - The distribution of Fe(III), its availability for microbial reduction, and factors controlling Fe(III) availability were investigated in sediments from a freshwater site in the Potomac River Estuary. Fe(III) reduction in sediments incubated under anaerobic conditions and depth profiles of oxalate-extractable Fe(III) indicated that Fe(III) reduction was limited to depths of 4 cm or less, with the most intense Fe(III) reduction in the top 1 cm. In incubations of the upper 4 cm of the sediments, Fe(III) reduction was as important as methane production as a pathway for anaerobic electron flow because of the high rates of Fe(III) reduction in the 0- to 0.5-cm interval. Most of the oxalate-extractable Fe(III) in the sediments was not reduced and persisted to a depth of at least 20 cm. The incomplete reduction was not the result of a lack of suitable electron donors. The oxalate-extractable Fe(III) that was preserved in the sediments was considered to be in a form other than amorphous Fe(III) oxyhydroxide, since synthetic amorphous Fe(III) oxyhydroxide, amorphous Fe(III) oxyhydroxide adsorbed onto clay, and amorphous Fe(III) oxyhydroxide saturated with adsorbed phosphate or fulvic acids were all readily reduced. Fe(3)O(4) and the mixed Fe(III)-Fe(II) compound(s) that were produced during the reduction of amorphous Fe(III) oxyhydroxide in an enrichment culture were oxalate extractable but were not reduced, suggesting that mixed Fe(III)-Fe(II) compounds might account for the persistence of oxalate-extractable Fe(III) in the sediments. The availability of microbially reducible Fe(III) in surficial sediments demonstrates that microbial Fe(III) reduction can be important to organic matter decomposition and iron geochemistry. However, the overall extent of microbial Fe(III) reduction is governed by the inability of microorganisms to reduce most of the Fe(III) in the sediment. PMID- 16347169 TI - Bacterial physiological diversity in the rhizosphere of range plants in response to retorted shale stress. AB - Bacterial populations were isolated from the soil-root interface and root-free regions of Agropyron smithii Rydb. and Atriplex canescens (Pursh) Nutt. grown in soil, retorted shale, or soil over shale. Bacteria isolated from retorted shale exhibited a wider range of tolerance to alkalinity and salinity and decreased growth on amino acid substrates compared with bacteria from soil and soil-over shale environments. Exoenzyme production was only slightly affected by growth medium treatment. Viable bacterial populations were higher in the rhizosphere and rhizoplane of plants grown in retorted shale than in plants grown in soil or soil over shale. In addition, a greater number of physiological groups of rhizosphere bacteria was observed in retorted shale compared with soil alone. Two patterns of community similarity were observed in comparisons of bacteria from soil over shale with those from soil and retorted-shale environments. Root-associated populations from soil over shale had a higher proportion of physiological groups in common with those from the soil control than with those from the retorted shale treatment. However, in non-rhizosphere populations, bacterial groups from soil over shale more closely resembled the physiological groups from retorted shale. PMID- 16347170 TI - Ultrastructure and Host Specificity of Bacteriophages of Streptococcus cremoris, Streptococcus lactis subsp. diacetylactis, and Leuconostoc cremoris from Finnish Fermented Milk "Viili". AB - "Viili," a fermented milk product, has a firm but viscous consistency. It is produced with traditional mesophilic mixed-strain starters, which have various stabilities in dairy practice. Thirteen morphologically different types of phages were found in 90 viili samples studied by electron microscopy. Ten of the phage types had isometric heads with long, noncontractile tails, two had elongated heads with long, noncontractile tails, and one had a unique, very long elongated head with a short tail. Further morphological differences were found in the tail size and in the presence or absence of a collar, a baseplate, and a tail fiber. To find hosts for the industrially significant phages, we examined the sensitivities of 500 bacterial isolates from starters of the viili. Seven of the phages attacked Streptococcus cremoris strains, three attacked S. lactis subsp. diacetylactis strains, and four attacked Leuconostoc cremoris strains. Some phages differed only in their host specificity. Hosts were not found for 4 of the 13 morphological types of phages. PMID- 16347171 TI - Inhibition of ammonium oxidation by nitrapyrin in soil and liquid culture. AB - Inhibition of growth of axenic cultures of Nitrosomonas europaea by nitrapyrin was investigated in liquid culture and in soil. In liquid culture, exponentially growing cells were more sensitive than stationary-phase cells, possibly due to a requirement for uptake of nitrapyrin, metabolism of nitrapyrin, or both before inhibition. Differences in sensitivity were observed between the parent strain and two strains, sp1 and sp2, that were selected through repeated subculturing. These differences were reflected in the length of the lag period induced by nitrapyrin and in the specific growth rate and were due to different bactericidal and bacteriostatic effects. Soil provided significant protection from inhibition, with concentrations of nitrapyrin approximately one order of magnitude greater than those required for equivalent inhibition in liquid culture. The data show that strain differences alone do not explain differences in sensitivity between nitrification in soil and in liquid culture and suggest that the inhibitor may be more effective against actively nitrifying soils. PMID- 16347172 TI - Acetylene metabolism and stimulation of denitrification in an agricultural soil. AB - The effects of C(2)H(2) metabolism on N(2)O production were examined in soil slurries. Enrichment of C(2)H(2) consumption activity occurred only in aerobic incubations. Rapid disappearance of subsequent C(2)H(2) additions, stimulation of CO(2) production, and most-probable-number enumerations of C(2)H(2) utilizers indicated enrichment of the population responsible. During C(2)H(2) consumption in slurries incubated statically under air, maximal rates of N(2)O evolution were 19 times higher than those in anaerobic incubations. After 20 days of enrichment with C(2)H(2), the production of N(2)O by slurries supplemented with C(2)H(2) and nitrate was 10 times higher than that in the unenriched controls. A Nocardia- or Arthrobacter-like bacterium was isolated that grew on C(2)H(2) but did not denitrify. The behavior of soil inoculated with this bacterium became similar to that of C(2)H(2)-enriched soil incubated aerobically. Ethanol, acetate, and acetaldehyde were identified in enrichment experiments, and denitrification in soil slurries was stimulated by addition of the supernatant from a pure culture grown on mineral medium with C(2)H(2). These results indicate that denitrification can be stimulated by the actions of an aerobic, nondenitrifying C(2)H(2)-metabolizing population. Utilization of intermediate metabolites by denitrifiers and enhanced O(2) consumption are two possible mechanisms for this stimulation. PMID- 16347173 TI - Bacterial Growth Rates and Competition Affect Nodulation and Root Colonization by Rhizobium meliloti. AB - The addition of streptomycin to nonsterile soil suppressed the numbers of bacterial cells in the rhizosphere of alfalfa (Medicago sativa L.) for several days, resulted in the enhanced growth of a streptomycin-resistant strain of Rhizobium meliloti, and increased the numbers of nodules on the alfalfa roots. A bacterial mixture inoculated into sterile soil inhibited the colonization of alfalfa roots by R. meliloti, caused a diminution in the number of nodules, and reduced plant growth. Enterobacter aerogenes, Pseudomonas marginalis, Acinetobacter sp., and Klebsiella pneumoniae suppressed the colonization by R. meliloti of roots grown on agar and reduced nodulation by R. meliloti, the suppression of nodulation being statistically significant for the first three species. Bradyrhizobium sp. and "Sarcina lutea" did not suppress root colonization nor nodulation by R. meliloti. The doubling times in the rhizosphere for E. aerogenes, P. marginalis, Acinetobacter sp., and K. pneumoniae were less and the doubling times for Bradyrhizobium sp. and "S. lutea" were greater than the doubling time of R. meliloti. Under the same conditions, Arthrobacter citreus injured alfalfa roots. We suggest that competition by soil bacteria reduces nodulation by rhizobia in soil and that the extent of inhibition is related to the growth rates of the rhizosphere bacteria. PMID- 16347174 TI - Thirteen Virulent and Temperate Bacteriophages of Lactobacillus bulgaricus and Lactobacillus lactis Belong to a Single DNA Homology Group. AB - Thirteen virulent phages and two temperate phages of two closely related bacterial species (Lactobacillus lactis and L. bulgaricus) were compared for their protein composition, their antigenic properties, their restriction endonuclease patterns, and their DNA homology. The immunoblotting studies and the DNA-DNA hybridizations showed that the phages could be differentiated into two groups. One group contained 2 temperate phages of L. bulgaricus and 11 virulent phages of L. lactis. Inside each group, at least two common proteins of identical sizes could be detected for each phage. These proteins were able to cross-react in immunoblotting experiments with an antiserum raised against one phage of the same group. Temperate phage DNAs showed partial homology with DNAs from some virulent phages. These homologies seem to be located on the region coding for the structural proteins since recombinant plasmids coding for one of the major phage proteins of one phage were able to hybridize with the DNAs from phages of the same group. These results suggest that temperate and virulent phages may be related to one another. PMID- 16347175 TI - Gram-negative versus gram-positive (actinomycete) nonobligate bacterial predators of bacteria in soil. AB - The existence of nonobligate bacterial predators of bacteria in soil has been previously reported. Several additional predators were isolated from soil and tested for predation in situ in soil by use of the indirect bacteriophage analysis technique. The trials were conducted with nutritionally poor and nutritionally enriched soil. Certain of the predators that were gram negative were found to attack a range of both gram-positive and gram-negative host cell species, including at least some of the other predator bacteria, both gram positive and gram negative. The attack occurred in both the nutritionally poor and rich soils, but in some instances it was somewhat depressed in the rich soil. This may be due to the nonobligate nature of the predation. The gram-positive predators attacked a relatively narrow range of prey species, and the attack occurred only in the nutritionally rich soil. In addition, the gram-positive predators were subject to attack by certain of the gram-negative predators. These gram-negative predators therefore appeared to play a dominant role in the control of bacterial numbers in soil. PMID- 16347176 TI - Large-Scale Production of Rhizobium meliloti on Whey. AB - Whey, a by-product of the cheese industry, can sustain the growth of fast-growing rhizobia. To avoid any latency of growth, rhizobial inoculum must be prepared under inducing conditions. In unsupplemented whey, the number of cells of Rhizobium meliloti Balsac reached 5 x 10 CFU/ml in 48 h of incubation. This is comparable to the yield obtained with yeast-mannitol broth, the standard medium for the growth of rhizobia. In raw whey supplemented with yeast extract (1.0 g/liter) and phosphate (0.5 g/liter), the number of cells reached 10 CFU/ml in 48 h of incubation. This is a twofold increase compared with the population normally obtained in industrial production. Whey represents a relatively inexpensive and efficient substrate medium for the large-scale production of fast-growing rhizobia. PMID- 16347177 TI - Rapid Colored-Nodule Assay for Assessing Root Exudate-Enhanced Competitiveness of Bradyrhizobium japonicum. AB - The effects of root exudate (RE) treatment on nodule occupancy by Bradyrhizobium japonicum were investigated by a rapid colored-nodule assay, which is based on the observation that B. japonicum L-110 and its antibiotically marked derivatives form dark-red nodules on certain soybean (Glycine max) cultivars, whereas other strains form beige nodules. The efficacy of the assay was confirmed by direct immunofluorescence and by antibiotic platings of nodule bacteria. Both logarithmic- and stationary-phase cultures of the reference strain, L-110Nal, were used in paired-competition studies with RE-treated or untreated cells of seven challenge strains. On the basis of field and greenhouse competition studies, these strains were placed into three competitiveness groups: high (AN 11, AN-16aStrRif, and AN-6), intermediate (AN-3 and 122SR), and low (I-110ARS and AN-18). Seedlings of G. max cv. Centennial were inoculated with two ratios of challenge to reference strain, 1:1 and 1:9, and nodule occupancy was determined after the V4 to V5 stage of ontogeny. Two of the strains showed increased occupancy in response to RE treatment at the 1:1 inoculation ratio. Logarithmic- and stationary-phase cultures of AN-6 showed increased occupancy, from 22 to 38% (P < 0.10) and from 23 59 39% (P < 0.05), respectively. While the maximum increase for stationary-phase cultures of AN-16aStrRif was from 34 to 47% (P < 0.05), logarithmic-phase cultures failed to respond to RE treatment. The results of these studies indicate that RE treatment increases the nodule occupancy of some, but not all, B. japonicum strains and that the colored-nodule assay could be rapidly and reliably used to determine the competitive ability of B. japonicum. PMID- 16347178 TI - Method for Assessing Heterogeneity in Turnover Rates within Microbial Communities. AB - A method is presented for determining both the average turnover rate and the standard deviation of the average turnover rate of the adenine nucleotide (AN) pool within a population of microorganisms. The method requires the calculation of the initial slope and curvature of a plot of AN specific activity versus time following the introduction of [H]adenine. An analysis of noise-corrupted data indicated that the method is capable of detecting a lack of uniformity in the turnover rate when the coefficient of variation of the turnover rate exceeds 39%. An analysis of field data revealed a significant lack of uniformity in the turnover rates of microbial communities in a marine sediment sample and freshwater pond but no significant nonuniformity in the turnover rates of microbial communities in a seawater sample and in a second freshwater pond. Although the method has been applied only to the analysis of AN turnover rates, it is applicable to any intracellular pool for which a suitable radioactive precursor exists. PMID- 16347179 TI - In Vitro Growth of Acremonium coenophialum, an Endophyte of Toxic Tall Fescue Grass. AB - Acremonium coenophialum, an endophytic fungus present in toxic tall fescue grass and seed, grew very slowly or not at all with conventional media and cultural practices. However, a considerable increase in growth was achieved in a relatively dilute medium consisting solely of glucose and yeast extract. The optimal levels of glucose and yeast extract were 3 to 6% and 0.35% (wt/vol), respectively. The addition of salts which lowered the pH suppressed growth. Even when the pH was controlled, the addition of KH(2)PO(4) at a level of 3.2% or more greatly inhibited growth. A. coenophialum grew better in shake culture than in stationary culture. The optimal temperature was 23 degrees C, and the optimal pH was 6.5. PMID- 16347180 TI - Synthesis of [C]Cellobiose with Clostridium thermocellum Cellobiose Phosphorylase. AB - Cellobiose labeled either at the reducing end (4-O-beta-d-glucopyranosyl-d-[U C]glucopyranose) or at the nonreducing end (4-O-beta-d-[U-C]glucopyranosyl-d glucopyranose) was synthesized with Clostridium thermocellum cellobiose phosphorylase at greater than 98% purity. The radioactive cellobioses were identical to authentic cellobiose in melting point, optical rotation, isotopic dilution, and chromatographic properties. PMID- 16347181 TI - Combined Determination of Poly-beta-Hydroxyalkanoic and Cellular Fatty Acids in Starved Marine Bacteria and Sewage Sludge by Gas Chromatography with Flame Ionization or Mass Spectrometry Detection. AB - Extraction of lipids from bacterial cells or sewage sludge samples followed by simple and rapid extraction procedures and room temperature esterification with pentafluorobenzylbromide allowed combined determinations of poly-beta hydroxyalkanoate constituents and fatty acids. Capillary gas chromatography and flame ionization or mass spectrometric detection was used. Flame ionization permitted determination with a coefficient of variation ranging from 10 to 27% at the picomolar level, whereas quantitative chemical ionization mass spectrometry afforded sensitivities for poly-beta-hydroxyalkanoate constituuents in the attomolar range. The latter technique suggests the possibility of measuring such components in bacterial assemblies with as few as 10 cells. With the described technique using flame ionization detection, it was possible to study the rapid formation of poly-beta-hydroxyalkanoate during feeding of a starved marine bacterium isolate with a complex medium or glucose and correlate the findings to changes in cell volumes. Mass spectrometric detection of short beta-hydroxy acids in activated sewage sludge revealed the presence of 3-hydroxybutyric, 3 hydroxyhexanoic, and 3-hydroxyoctanoic acids in the relative proportions of 56, 5 and 39%, respectively. No odd-chain beta-hydroxy acids were found. PMID- 16347182 TI - Fractionation and Estimation of Particle-Attached and Unattached Bradyrhizobium japonicum Strains in Soils. AB - Rhizobial cells attached or unattached to soil particles were estimated. Nonsterile soils into which antibiotic-resistant mutants of Bradyrhizobium japonicum had been introduced were fractionated by a centrifugation technique into two fractions: A, which contained mainly rhizobial cells attached to soil particles, and F, which contained mainly rhizobial cells unattached to them. Rhizobial counts decreased in both fractions during incubation of the soil at 30 degrees C, with a concomitant decrease in the proportion of the count of fraction F to that of fraction A. Sonication of fraction A of the soil incubated for more than 3 weeks caused an increase in the rhizobial count. The ratio of the count of fraction A estimated by the plant infection method to that estimated by the dilution plate method increased after 5 days of soil incubation. More than 90% of the indigenous rhizobia in an agricultural field existed in fraction A. These results suggest that the majority of rhizobial cells are attached to soil particles. PMID- 16347183 TI - Alteration of a salt marsh bacterial community by fertilization with sewage sludge. AB - The effects of long-term fertilization with sewage sludge on the aerobic, chemoheterotrophic portion of a salt marsh bacterial community were examined. The study site in the Great Sippewissett Marsh, Cape Cod, Mass., consisted of experimental plots that were treated with different amounts of commercial sewage sludge fertilizer or with urea and phosphate. The number of CFUs, percentage of mercury- and cadmium-resistant bacteria, and percentage of antibiotic-resistant bacteria were all increased in the sludge-fertilized plots. Preliminary taxonomic characterization showed that sludge fertilization markedly altered the taxonomic distribution and reduced diversity within both the total heterotrophic and the mercury-resistant communities. In control plots, the total heterotrophic community was fairly evenly distributed among taxa and the mercury-resistant community was dominated by Pseudomonas spp. In sludge-fertilized plots, both the total and mercury-resistant communities were dominated by a single Cytophaga sp. PMID- 16347184 TI - Isolation and Characterization of Linear DNA Elements from the Mitochondria of Gaeumannomyces graminis. AB - Different Gaeumannomyces graminis strains of diverse geographic origin contain one or two small DNAs ranging in size from 7.2 to 10 kilobases. These DNAs exhibit different degrees of homology with each other. We have characterized these low-molecular-weight DNAs from one strain, Ha-01. These small DNAs, E1 and E2, are mitochondrial in origin and were isolated as linear molecules which exhibited an intrinsic difference in density from the high-molecular-weight DNA. PMID- 16347185 TI - Effects of flow cytometric analysis and cell sorting on photosynthetic carbon uptake by phytoplankton in cultures and from natural populations. AB - Photosynthetic rates of phytoplankton were significantly lower after analysis by flow cytometry (FCM) than before. Exposure to the laser beam during the sorting process caused significant physiological damage. The cellular content of a radiolabel accumulated prior to FCM was not affected by FCM. Although it may not be possible to use FCM to preconcentrate cells for further physiological studies, samples may be incubated with stable or radioactive isotopes and then analyzed by FCM. PMID- 16347186 TI - Plasmid Involvement in Linalool Metabolism by Pseudomonas fluorescens. AB - A bacterial strain was isolated from a wastewater lagoon and identified as Pseudomonas fluorescens. This isolate was able to utilize linalool as a sole carbon and energy source. The ability was found to be encoded on a 60-megadalton transmissible plasmid, pSRQ60. The plasmid was also mated into a commercial waste treatment strain, which expanded its ability to utilize other isoprenoid compounds. PMID- 16347187 TI - Selection of yeasts for breadmaking by the frozen-dough method. AB - Eleven yeast strains suitable for frozen dough were selected from over 300 Saccharomyces strains. All of these were identified as Saccharomyces cerevisiae from morphological, cultural, and physiological characteristics. The selected yeast cells accumulated a higher amount of trehalose than did commercial bakers' yeast cells. PMID- 16347188 TI - Rapid microbial mineralization of toluene and 1,3-dimethylbenzene in the absence of molecular oxygen. AB - Up to 0.4 mM 1,3-dimethylbenzene (m-xylene) was rapidly mineralized in a laboratory aquifer column operated in the absence of molecular oxygen with nitrate as an electron acceptor. Under continuous flow conditions, the degradation rate constant (pseudo-first order) was >0.45 h. Based on a carbon mass balance with [ring-C]m-xylene and a calculation of the electron balance, m xylene was shown to be quantitatively (80%) oxidized to CO(2) with a concomitant reduction of nitrate. The mineralization of m-xylene in the column also took place after reducing the redox potential, E', of the inflowing medium with sulfide to <-0.11 V. Microorganisms adapted to growth on m-xylene were also able to degrade toluene under denitrifying conditions. These results suggest that aromatic hydrocarbons present in anoxic environments such as lake sediments, sludge digestors, and groundwater infiltration zones from landfills and polluted rivers are not necessarily persistent but may be mineralized in the absence of molecular oxygen. PMID- 16347189 TI - Comparison of bacterial lipopolysaccharides by high-performance liquid chromatography. AB - A comparison of lipid-free polysaccharides from gram-negative bacteria was rapidly accomplished by using high-performance liquid chromatography of underivatized hydrolysates. Examination of a number of such products revealed that, contrary to earlier reports, Xanthomonas campestris lipopolysaccharide contained heptose, together with rhamnose and galactose, but not mannose. The polymers from the methanotrophs "Methylomonas albus" and "Methylosinus trichosporium" contained heptose and glucose, and that from a "Klebsiella aerogenes" strain contained heptose, glucose, and galactose. The absence of heptose from the lipopolysaccharide of Myxococcus xanthus was confirmed. PMID- 16347190 TI - Growth of fast- and slow-growing rhizobia on ethanol. AB - Free-living soybean rhizobia and Bradyrhizobium spp. (lupine) have the ability to catabolize ethanol. Of the 30 strains of rhizobia examined, only the fast- and slow-growing soybean rhizobia and the slow-growing Bradyrhizobium sp. (lupine) were capable of using ethanol as a sole source of carbon and energy for growth. Two strains from each of the other Rhizobium species examined (R. meliloti, R. loti, and R. leguminosarum biovars phaseoli, trifolii, and viceae) failed to grow on ethanol. One Rhizobium fredii (fast-growing) strain, USDA 191, and one (slow growing) Bradyrhizobium japonicum strain, USDA 110, grew in ethanol up to concentrations of 3.0 and 1.0%, respectively. While three of the R. fredii strains examined (USDA 192, USDA 194, and USDA 205) utilized 0.2% acetate, only USDA 192 utilized 0.1% n-propanol. None of the three strains utilized 0.1% methanol, formate, or n-butanol as the sole carbon source. PMID- 16347191 TI - Semienclosed Tube Cultures of Bean Plants (Phaseolus vulgaris L.) for Enumeration of Rhizobium phaseoli by the Most-Probable-Number Technique. AB - The semienclosed tube culture technique of Gibson was modified to permit growth of common bean (Phaseolus vulgaris L.) roots in humid air, enabling enumeration of the homologous (nodule forming) symbiont, Rhizobium phaseoli, by the most probable-number plant infection method. A bean genotype with improved nodulation characteristics was used as the plant host. This method of enumeration was accurate when tubes were scored 3 weeks after inoculation with several R. phaseoli strains diluted from aqueous suspensions, peat-based inoculants, or soil. A comparison of population sizes obtained by most-probable-number tube cultures and plate counts indicated that 1 to 3 viable cells of R. phaseoli were a sufficient inoculant to induce nodule formation. PMID- 16347192 TI - Dinitrogen production from nitrite by a nitrosomonas isolate. AB - A chemolithotrophic ammonium-oxidizing bacterium that was able to reduce NO(2) to N(2) (m/z 30) while oxidizing ammonium under conditions of oxygen stress was isolated from stream sediments. Energy was derived from ammonium oxidation, as evidence by growth, with CO(2) serving as the sole C source. The organism was a gram-negative, motile, short rod that failed to grow either aerobically or anaerobically in heterotroph media. The organism was identified as a Nitrosomonas sp. PMID- 16347193 TI - Effects of inoculum size on solid-phase fermentation of fodder beets for fuel ethanol production. AB - This fuel ethanol study examined the effects of Saccharomyces cerevisiae inoculum size on solid-phase fermentation of fodder beet pulp. A 5% inoculum (wt/wt) resulted in rapid yeast and ethanol (9.1% [vol/vol]) production. Higher inocula showed no advantages. Lower inocula resulted in lowered final yeast populations and increased fermentation times. PMID- 16347194 TI - Inhibitor Studies of Dissimilative Fe(III) Reduction by Pseudomonas sp. Strain 200 ("Pseudomonas ferrireductans"). AB - [This corrects the article on p. 383 in vol. 52.]. PMID- 16347195 TI - Electrostatic Mechanism of Survival of Virulent Aeromonas salmonicida Strains in River Water. AB - [This corrects the article on p. 1343 in vol. 51.]. PMID- 16347196 TI - Conjugal strategy for construction of fast Acid-producing, bacteriophage resistant lactic streptococci for use in dairy fermentations. AB - Bacteriophage-resistant dairy streptococci were obtained following conjugal transfer of pTR2030 from a lactose-negative donor, Streptococcus lactis TEK12, to lactose-positive recipient strains, Streptococcus cremoris LMA13 and 924 and S. lactis LMA12. Fast acid-producing, phage-resistant transconjugants were selected by challenge with homologous phage on fast-slow differential agar or lactose indicator agar. Acquisition of pTR2030 by the transconjugants was confirmed by DNA-DNA hybridization. Resistance of transconjugants to homologous phage was complete. Curing or deletion of pTR2030 in the transconjugants confirmed that phage resistance was due to pTR2030 acquisition and not to coincident background mutation. Phage-sensitive pTR2030 deletion derivatives of LMA12 transconjugants were isolated in vivo. The HindIII fragment B of pTR2030 was subcloned into pBR322 to yield a recombinant plasmid, pMET2, useful as a source of pTR2030 DNA. A specific, chemically synthesized oligomer useful as a pTR2030 probe was derived from the sequence of a small portion of pTR2030. The conjugal strategy presented here was effective in yielding fast acid-producing, phage-resistant S. cremoris and S. lactis strains without the use of antibiotic resistance markers and without interfering with the acid-producing ability of the recipient strain. PMID- 16347197 TI - Enrichment for Hydrogen-Oxidizing Acinetobacter spp. in the Rhizosphere of Hydrogen-Evolving Soybean Root Nodules. AB - Field soybean plants were inoculated with Hup wild-type or H(2) uptake-negative (Hup) mutants of Bradyrhizobium japonicum. For two consecutive summers we found an enrichment for acinetobacters associated with the surfaces of the H(2) evolving nodules. Soybean root nodules that evolved H(2) had up to 12 times more Acinetobacter spp. bacteria associated with their surfaces than did nodules incapable of evolving H(2). All of the newly isolated strains identified as Acinetobacter obtained from the surfaces of root nodules, as well as known established Acinetobacter strains, were capable of oxidizing H(2), a property not previously described for this alkane-degrading soil bacterium. PMID- 16347198 TI - Autecology in Rhizospheres and Nodulating Behavior of Indigenous Rhizobium trifolii. AB - Indigenous serotype 1-01 of Rhizobium trifolii occupied significantly fewer nodules (6%) on plants of soil-grown noninoculated subterranean clover (Trifolium subterraneum L.) cv. Woogenellup than on cv. Mt. Barker (36%) sampled at the flowering stage of growth. Occupancy by indigenous serotype 2-01, was not significantly different on the two cultivars (16 and 26%). Serotype-specific, fluorescent-antibody conjugates were synthesized and used to enumerate the indigenous serotypes in host (clovers) and nonhost (annual rye-grass, Lolium multiflorum L.) rhizospheres and in nonplanted soil. The form and concentration of Ca in the flocculating mixture and the presence of phosphate anions in the extracting solution were both critical for enumerating R. trifolii in Whobrey soil. The two serotypes were present in similar numbers in nonplanted soil (ca. 10 per g of soil) and each represented ca. 10% of the total R. trifolii population. Although host rhizospheres did not preferentially stimulate either serotype, the mean population densities of serotype 2-01 were significantly greater (P = 0.05) than those of serotype 1-01 in clover rhizospheres on 8 of 14 samplings made between the time of seeding and the appearance of nodules (day 12). In this experiment, and in contrast to our earlier findings, serotype 1-01 occupied significantly fewer (P 90% of the nodules on both cultivars were occupied by the more abundant strain in the inoculum regardless of sampling date (4 or 8 weeks). In contrast, large percentages of nodules on 4-week-old plants of both cultivars exposed to a 5:1 inoculum mixture were doubly occupied (64 and 74%). By week 8 these values had decreased significantly (P C(2)H(4)) in Azolla filiculoides and on the nitrogen fixation and growth of free-living Anabaena azollae 2B organisms. Inoculation of azolla medium with Chlamydomonas sp. was associated with decreased nitrogenase activity in A. filiculoides and with increases in the density of a fungal population identified as Acremonium sp. Subsequent inoculation of azolla medium with this fungus was also accompanied by a significant decrease in nitrogenase activity of A. filiculoides. However, the extent of depression of nitrogenase activity was significantly higher when azolla medium was inoculated with Chlamydomonas sp. than when it was inoculated with Acremonium sp. Inoculation of nitrogen-free Stanier medium with either Acremonium sp. or Chlamydomonas sp. did not adversely affect the growth or nitrogenase activity of free-living A. azollae. Decreased nitrogenase activity in A. filiculoides is apparently related to the adverse influence of the green alga and the fungus on the macrosymbiont. The mechanisms that might be involved are discussed. PMID- 16347212 TI - Evaluation of Diverse Antisera, Conjugates, and Support Media for Detecting Bradyrhizobium japonicum by Indirect Enzyme-Linked Immunosorbent Assay. AB - We evaluated three antisera and four enzyme conjugates for the detection of Bradyrhizobium japonicum by an indirect enzyme-linked immunosorbent assay in microtiter plates. Nitrocellulose membrane sheets were then evaluated as an alternative support medium by using some combinations. Partially purified immunoglobulin G (IgG) or unpurified antisera to strain USDA 110 raised in rabbits, goats, or sheep was reacted in microtiter plates with alkaline phosphatase conjugated to protein A, goat anti-rabbit (GAR), sheep anti-rabbit (SAR), or rabbit anti-goat (RAG) IgG. Cultures or nodules containing homologous rhizobia were detected with equal sensitivity when protein A, GAR, or SAR was reacted with 5 mug of protein IgG per ml or a 1:800 titer of antisera from rabbits, but not goats or sheep. RAG reacted with IgG or antisera from goats or sheep. The detection limit was 2 x 10 rhizobia per well. Rhizobia were spotted on nitrocellulose sheets as an alternative support medium, followed by soaking in 5 mug of protein per ml as IgG and 1:4,000 dilutions of protein A or GAR conjugate. Rhizobia in serogroup 110 were detected with the dye combination Nitro Blue Tetrazolium-5-bromo-4-chloro-3-indolyl phosphate (NBT-BCIP), and rhizobia in serogroup 122 were detected with fast red-naphthol phosphate (FR-NP). At the conclusion of the 5-h assay, purple (NBT-BCIP) or red (FR-NP) spots were visible in positive reactions. The sensitivity of detection was about 1,000 rhizobial cells or 3 mug of nodules tissue. PMID- 16347213 TI - Structure and dynamics of anaerobic bacterial aggregates in a gas-lift reactor. AB - Anaerobic mixed-culture aggregates, which converted glucose to acetic, propionic, butyric, and valeric acids, were formed under controlled conditions of substrate feed (carbon limitation) and hydraulic regimen. The continuous-flow system used (anaerobic gas-lift reactor) was designed to retain bacterial aggregates in a well-mixed reactor. Carrier availability (i.e., liquid-suspended sand grains) proved necessary for bacterial aggregate formation from individual cells during reactor start-up. Electron microscopic examination revealed that incipient colonization of sand grains by bacteria from the bulk liquid occurred in surface irregularities, conceivably reflecting local quiescence. Subsequent confluent biofilm formation on sand grains proved to be unstable, however. Substrate depletion in the bulk liquid is assumed to weaken deeper parts of the biofilm due to cellular lysis, after which production of gas bubbles and liquid shearing forces cause sloughing. The resulting fragments, although sand free, were nevertheless large enough to be retained in the reactor and gradually grew larger through bacterial growth and by clumping together with other fragments. In the final steady state, high cell densities were maintained in the form of aggregates, while sand had virtually disappeared due to sampling losses and wash out. Numerical cell densities within aggregates ranged from 10/ml at the periphery to very low values in the center. The cells were enmeshed in a polymer matrix containing polysaccharides; nevertheless, carbon sufficiency was not a prerequisite to sustain high hold-up ratios. PMID- 16347214 TI - Production, Purification, and Characterization of alpha-Galactosidase from Monascus pilosus. AB - A Monascus pilosus strain was selected for production of intracellular alpha galactosidase. Optimum conditions for mycelial growth and enzyme induction were determined. Galactose was one of the best enzyme inducers. The enzyme was purified by ammonium sulfate precipitation, gel filtration, and ion exchange chromatography and was demonstrated to be homogeneous by slab gel electrophoresis. The molecular weight of this enzyme, estimated by gel filtration, was about 150,000. The optimum conditions for the enzyme reaction was pH 4.5 to 5.0 at 55 degrees C. The purified enzyme was stable at 55 degrees C or below and in buffer at pH 3 to 8. The activity was inhibited by mercury, silver, and copper ions. The kinetics of this enzyme, with p-nitrophenyl-alpha-d galactoside as substrate, was determined: K(m) was about 0.8 mM, and V(max) was 39 mumol/min per mg of protein. Enzymatic hydrolysis of melibiose, raffinose, and stachyose was analyzed by thin-layer chromatography. PMID- 16347215 TI - Comparative Study of Action of Cell Wall Proteinases from Various Strains of Streptococcus cremoris on Bovine alpha(s1)-, beta-, and kappa-Casein. AB - Partially purified cell wall proteinases of eight strains of Streptococcus cremoris were compared in their action on bovine alpha(s1)-, beta-, and kappa casein, as visualized by starch gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and thin-layer chromatography. Characteristic degradation profiles could be distinguished, from which the occurrence of two proteinases, represented by strain HP and strain AM(1), was concluded. The action of the HP-type proteinase P(1) (also detectable in strains Wg(2), C(13), and TR) was established by electrophoretic methods to be directed preferentially towards beta-casein. The AM(1)-type proteinase P(III) (also detectable in strain SK(11)) was also able to degrade beta-casein, but at the same time split alpha(s1)- and kappa-casein more extensively than did P(I). Strain FD(27) exhibited mainly P(I) activity but also detectable P(III) degradation characteristics. The cell wall proteinase preparation of strain E(8) showed low P(I) as well as low P(III) activity. All proteinase preparations produced from kappa-casein positively charged degradation products with electrophoretic mobilities similar to those of degradation products released by the action of the milk-clotting enzyme chymosin. The differences between P(I) and P(III) in mode of action, as detected by gel electrophoresis and thin-layer chromatography, were reflected by the courses of the initial degradation of methyl-C-labeled beta-casein and by the effect of alpha(s1)- plus kappa-casein on these degradations. The results are discussed in the light of previous comparative studies of cell wall proteinases in strains of S. cremoris and with respect to the growth of this organism in milk. PMID- 16347216 TI - Microbial iron reduction by enrichment cultures isolated from estuarine sediments. AB - Microbial Fe reduction in acetate- and succinate-containing enrichment cultures initiated with an estuarine sediment inoculum was studied. Fe reduction was unaffected when SO(4) reduction was inhibited by MoO(4), indicating that both processes could occur independently. Bacterially produced sulfide precipitated as FeS but was not completely responsible for Fe reduction. The separation of oxidized Fe particles from bacteria by dialysis tubing demonstrated that direct bacterial contact was necessary for Fe reduction. Fe reduction in cultures amended with NO(3) was delayed until NO(3) and NO(2) were removed. However, bacterial attachment to oxidized Fe particles in NO(3)-amended cultures occurred early during growth in a manner similar to NO(3)-free cultures. During late stages of growth, bacteria not attached to Fe particles became pale and swollen, while attached cells remained bright blue when examined by 4',6-diamidine-2 phenylindole epifluo-rescence microscopy. The presence of added oxidized Mn had no effect on Fe reduction. The results suggested that enzymatic Fe reduction was responsible for reducing Fe in these cultures even in the presence of sulfide and that cells incapable of Fe reduction became unhealthy when Fe(III) was the only available electron acceptor. PMID- 16347217 TI - Isolation of Pathogenic Bacillus circulans from Callus Cultures and Healthy Offshoots of Date Palm (Phoenix dactylifera L.). AB - Pure cultures of a gram-negative, rod-shaped bacterium, which produced endospores after 3 to 5 days on solid medium, were isolated exclusively from tissue cultures of the date palm Phoenix dactylifera L. Electron microscopic examination of thin sections of the bacteria revealed the bilayer membrane typical of gramnegative bacteria and confirmed the nature of the spores as true endospores. Biochemical and physiological tests indicated that the bacteria were Bacillus circulans. B. circulans was consistently isolated from the internal tissues, including the meristem, of apparently healthy offshoots of date palm. When meristem and embryo callus tissue culture samples were injected with B. circulans isolated from similar tissue culture samples and from offshoots, the majority of the isolates produced a rapid, destructive soft rot of the tissues. PMID- 16347218 TI - Stimulation of bacterial DNA synthesis by algal exudates in attached algal bacterial consortia. AB - Algal-bacterial consortia attached to polystyrene surfaces were prepared in the laboratory by using the marine diatom Amphora coffeaeformis and the marine bacterium Vibrio proteolytica (the approved name of this bacterium is Vibrio proteolyticus [W. E. C. Moore, E. P. Cato, and L. V. H. Moore, Int. J. Syst. Bacteriol. 35:382-407, 1985]). The organisms were attached to the surfaces at cell densities of approximately 5 x 10 cells cm (diatoms) and 5 x 10 cells cm (bacteria). The algal-bacterial consortia consistently exhibited higher rates of [H]thymidine incorporation than did biofilms composed solely of bacteria. The rates of [H]thymidine incorporation by the algal-bacterial consortia were fourfold greater than the rates of incorporation by monobacterial biofilms 16 h after biofilm formation and were 16-fold greater 70 h after biofilm formation. Extracellular material released from the attached Amphora cells supported rates of bacterial activity (0.8 x 10 to 17.9 x 10 mol of [H]thymidine incorporated cell h) and growth (doubling time, 29.5 to 1.4 days) comparable to values reported for a wide variety of marine and freshwater ecosystems. In the presence of sessile diatom populations, DNA synthesis by attached V. proteolytica cells was light dependent and increased with increasing algal abundance. The metabolic activity of diatoms thus appears to be the rate-limiting process in biofilm development on illuminated surfaces under conditions of low bulk-water dissolved organic carbon. PMID- 16347219 TI - Bacteriocinlike Activity within the Genus Thermus. AB - Members of the genus Thermus were examined for the presence of bacteriocinlike inhibitory activity. Testing was done by the deferred antagonism technique. Antagonistic activity, as evidenced by zones of inhibition, was expressed by Thermus rubens against all other Thermus strains tested. T. rubens itself was immune to this activity. Plasmid analysis of T. rubens revealed the presence of one plasmid of approximately 64 megadaltons. PMID- 16347220 TI - Thymidine Incorporation by the Microbial Community of Standing Dead Spartina alterniflora. AB - Thymidine incorporation by the microbial community on standing dead leaves of Spartina alterniflora did not obey many of the assumptions inherent in the use of the technique in planktonic systems. Incorporation rates of [methly-H]thymidine were nonsaturable over a wide concentration range (10 to 10 nM). Owing to metabolism by both fungi and bacteria, a major fraction of the radiolabel (mean, 48%) appeared in protein. Extraction of the radiolabeled macromolecules was inefficient, averaging 8.8%. Based on an empirically derived conversion factor, 4 x 10 cells . mol of thymidine, doubling times ranged from 4 to 69 h for the epiphytic bacterial assemblage. PMID- 16347221 TI - Further Verification of the Isotope Dilution Approach for Estimating the Degree of Participation of [H]thymidine in DNA Synthesis in Studies of Aquatic Bacterial Production. AB - The optimal concentration of [H]thymidine (i.e., the maximal degree of participation in DNA synthesis) as determined by adding increasing amounts of labeled thymidine at the same specific activity was similar to the concentration of thymidine inhibiting the de novo pathway as determined by isotope dilution plots. These experiments provide further verification of the isotope dilution approach for determining the degree of participation of [H]thymidine in DNA synthesis. PMID- 16347222 TI - Production and Rheological Properties of the Extracellular Polysaccharide Synthesized by Pseudomonas sp. Strain EPS-5028. AB - During batch aerobic submerged fermentation, the exopolysaccharide synthesis by Pseudomonas sp. strain EPS-5028 occurred in growth- and non-growth-linked processes. Polysaccharide formation increased when the pH was controlled at 7 during fermentation. Exopolysaccharide production depended on the phosphate content of the medium. The polymer exhibited a pseudoplastic nature, had good thermostability, and was affected neither by pH nor by high concentrations of salt. PMID- 16347223 TI - Lysozyme-sensitive bioemulsifier for immiscible organophosphorus pesticides. AB - Two Bacillus strains capable of emulsifying immiscible organophosphorus pesticides were isolated by enrichment methods. The emulsifying factor produced by Bacillus strain FE-2 has a high molecular weight, is Iysozyme sensitive and thermostable, and can be precipitated with trichloroacetic acid or ammonium sulfate; it may be a glycolipopeptide. It is specific for immiscible organophosphorus pesticides and is secreted during growth in the presence of such pesticides. PMID- 16347224 TI - Isolation and characterization of a fast-growing, thermophilic methanobacterium species. AB - A thermophilic, autotrophic methanogen (strain CB12, DSM 3664) was isolated from a mesophilic biogas digestor. This bacterium used H(2)-CO(2) or formate as a substrate and grew as short rods, sometimes in pairs and in crooked filaments. Motility was not observed. Its optimum temperature (56 degrees C) was lower than that of other thermophilic members of the genus Methanobacterium. The maximum observed specific growth rate was 0.564 h (74-min doubling time). PMID- 16347227 TI - Growth characteristics of a novel nitrogen-fixing cellulolytic bacterium. AB - Growth characteristics of a cellulolytic nitrogen-fixing bacterium isolated from a marine shipworm by Waterbury et al. (J. B. Waterbury, C. B. Calloway, and R. D. Turner, Science 221:1401-1403, 1983) are described. When grown microaerobically, the bacterium exhibited doubling times of about 2 days in cellulose-supplemented synthetic medium devoid of combined nitrogen. Maximum growth was reached 12 to 16 days after inoculation. Growth optima for pH, temperature, and NaCl concentration were 8.5, 30 to 35 degrees C, and 0.3 M, respectively. During growth the bacterium produced succinic acid (0.026%) and acetic acid (0.010%). Formic acid (0.010%) was produced during the stationary growth phase. No growth was observed when glucose was the sole carbon source. Cellobiose supported weak growth, while longer-chain-length cellodextrins supported extensive growth. Analysis of residual carbohydrates in the medium during growth indicated that the bacterium catabolized a terminal glucose moiety from the cellodextrin chain. PMID- 16347228 TI - Denitrification in marl and peat sediments in the Florida everglades. AB - The potential for denitrification in marl and peat sediments in the Shark River Slough in the Everglades National Park was determined by the acetylene blockage assay. The influence of nitrate concentration on denitrification rate and N(2)O yield from added nitrate was examined. The effects of added glucose and phosphate and of temperature on the denitrification potential were determined. The sediments readily denitrified added nitrate. N(2)O was released from the sediments both with and without added acetylene. The marl sediments had higher rates than the peat on every date sampled. Denitrification was nitrate limited; however, the yields of N(2)O amounted to only 10 to 34% of the added nitrate when 100 muM nitrate was added. On the basis of measured increases in ammonium concentration, it appears that the balance of added nitrate may be converted to ammonium in the marl sediment. The sediment temperature at the time of sampling greatly influenced the denitrification potential (15-fold rate change) at the marl site, indicating that either the number or the specific activity of the denitrifiers changed in response to temperature fluctuations (9 to 25 degrees C) in the sediment. It is apparent from this study that denitrification in Everglades sediments is not an effective means of removing excess nitrogen which may be introduced as nitrate into the ecosystem with supply water from the South Florida watershed and that sporadic addition of nitrate-rich water may lead to nitrous oxide release from these wetlands. PMID- 16347230 TI - Inhibitor Evaluation with Immobilized Nitrobacter agilis Cells. AB - Nitrobacter agilis was entrapped in calcium alginate beads and used as a floating bed supplied with a continuous flow of nitrite medium. Complete nitrite oxidation was achieved within 30 h, and the system could be maintained for at least 210 h. The immobilized Nitrobacter system was subjected to sulfur oxyanions, acidity, and metal ions. Thiosulfate and tetrathionate (up to 20 mM each) did not inhibit the nitrite oxidation activity. A low pH of 4.2 resulted in the complete cessation of nitrite oxidation, and the activity was not restored upon increasing the pH to 7. Nitrite oxidation by N. agilis was sensitive to 10 mM each Ni and Al but insensitive to 10 mM MoO(4). PMID- 16347229 TI - Depth distribution of bacterial production in a stratified lake with an anoxic hypolimnion. AB - The purpose of this study was to determine the depth distribution of bacterial biomass and production in a stratified lake and to test techniques to measure bacterial production in anaerobic waters. Bacterial abundance and incorporation of both [H]thymidine and [H]leucine into protein were highest in the metalimnion, at the depth at which oxygen first became unmeasurable. In contrast, [H]thymidine incorporation into DNA was highest in the epilimnion. The ratios of incorporation into DNA/protein averaged 2.2, 0.49, and 0.95 for the epilimnion, metalimnion, and hypolimnion, respectively. Low incorporation into DNA was not due to artifacts associated with the DNA isolation procedure. Recovery of added [H]DNA was about 90% in waters in which the portion of [H]thymidine incorporation into DNA was about 40%. At least some obligate anaerobic bacteria were capable of assimilating thymidine since aeration of anaerobic hypolimnion waters substantially inhibited thymidine incorporation. The depth profile of bacterial production estimated from total thymidine and leucine incorporation and the frequency of dividing cells were all similar, with maximal rates in the metalimnion. However, estimates of bacterial production based on frequency of dividing cells and leucine incorporation were usually significantly higher than estimates based on thymidine incorporation (using conversion factors from the literature), especially in anaerobic hypolimnion waters. These data indicate that the thymidine approach must be examined carefully if it is to be applied to aquatic systems with low oxygen concentrations. Our results also indicate that the interface between the aerobic epilimnion and anaerobic hypolimnion is the site of intense bacterial mineralization and biomass production which deserves further study. PMID- 16347231 TI - Factors affecting high-oxygen survival of heterotrophic microorganisms from an antarctic lake. AB - We sought to determine factors relating to the survival of heterotrophic microorganisms from the high-dissolved-oxygen (HDO) waters of Lake Hoare, Antarctica. This lake contains perpetual HDO about three times that of normal saturation (40 to 50 mg liter). Five isolates, one yeast and four bacteria, were selected from Lake Hoare waters by growth with the membrane filter technique with oxygen added to yield dissolved concentrations 14 times that in situ, 175 mg liter. One bacterial isolate was obtained from the microbial mat beneath the HDO waters. This organism was isolated at normal atmospheric oxygen saturation. The bacteria were gram-negative rods, motile, oxidase positive, catalase positive, and superoxide dismutase positive; they contained carotenoids. The planktonic isolates grew in media containing 10 mg of Trypticase soy (BBL Microbiology Systems)-peptone (2:1) liter but not at 10 g liter. Under low-nutrient levels simulating Lake Hoare waters (10 mg liter), two of the planktonic isolates tested were not inhibited by HDO. Growth inhibition by HDO increased as nutrient concentration was increased. A carotenoid-negative mutant of one isolate demonstrated a decreased growth rate, maximal cell density, and increased cell lysis in the death phase under HDO compared with the parent strain. The specific activity of superoxide dismutase was increased by HDO in four of the five bacterial isolates. The superoxide dismutase was of the manganese type on the basis of inhibition and electrophoretic studies. The bacterial isolates from Lake Hoare possess several adaptations which may aid their survival in the HDO waters, as well as protection due to the oligotrophic nature of the lake. PMID- 16347232 TI - Fixation, counting, and manipulation of heterotrophic nanoflagellates. AB - Quantitative effects of several fixatives on heterotrophic nanoflagellates (HNAN) and phototrophic nanoflagellates (PNAN) were investigated by hemacytometer and epifluorescence counting techniques. Counts of Monas sp. cultures before and after fixation with unbuffered 0.3% glutaraldehyde and 5% formaldehyde showed no loss of cells during fixation, and cell concentrations remained constant for several weeks after fixation. Buffering of fixatives with borax caused severe losses, up to 100% within 2 h. Field samples from Lake Vechten showed no decline of HNAN and total nanoflagellate concentrations for at least 1 week after fixation with 5% formaldehyde and with 1% glutaraldehyde. With 1% glutaraldehyde, the chlorophyll autofluorescence of PNAN was much brighter than with 5% formaldehyde, although it was lost after a few days and thus limited the storage time of samples. However, when primulin-stained slides were prepared soon after fixation and stored at -30 degrees C, the loss of autofluorescence was prevented and PNAN and HNAN concentrations were stable for at least 16 weeks. Effects of filtration and centrifugation on HNAN were also studied. Filtration vacuum could not exceed 3 kPa since 10 kPa already caused losses of 15 to 20%. Similar losses were caused by centrifugation, even at low speed (500 x g). PMID- 16347233 TI - Distinguishing between Nitrification and Denitrification as Sources of Gaseous Nitrogen Production in Soil. AB - The source of N(2)O produced in soil is often uncertain because denitrification and nitrification can occur simultaneously in the same soil aggregate. A technique which exploits the differential sensitivity of these processes to C(2)H(2) inhibition is proposed for distinguishing among gaseous N losses from soils. Denitrification N(2)O was estimated from 24-h laboratory incubations in which nitrification was inhibited by 10-Pa C(2)H(2). Nitrification N(2)O was estimated from the difference between N(2)O production under no C(2)H(2) and that determined for denitrification. Denitrification N(2) was estimated from the difference between N(2)O production under 10-kPa C(2)H(2) and that under 10 Pa. Laboratory estimates of N(2)O production were significantly correlated with in situ N(2)O diffusion measurements made during a 10-month period in two forested watersheds. Nitrous oxide production from nitrification was most important on well-drained sites of a disturbed watershed where ambient NO(3) was high. In contrast, denitrification N(2)O was most important on poorly drained sites near the stream of the same watershed. Distinction between N(2)O production from nitrification and denitrification was corroborated by correlations between denitrification N(2)O and water-filled pore space and between nitrification N(2)O and ambient NO(3). This technique permits qualitative study of environmental parameters that regulate gaseous N losses via denitrification and nitrification. PMID- 16347234 TI - Environmental Parameters Regulating Gaseous Nitrogen Losses from Two Forested Ecosystems via Nitrification and Denitrification. AB - Gaseous N losses from disturbed and reference forested watersheds at the Coweeta Hydrologic Laboratory in western North Carolina were studied by in situ N(2)O diffusion measurements and laboratory incubations throughout a 10-month period. Soil temperature, percent base saturation, and water-filled pore space accounted for 43% of the variation in in situ N(2)O diffusion measurements. Laboratory incubations distinguished the gaseous N products of nitrification and denitrification. Nitrifying activity, ambient NO(3), and nitrification N(2)O were positively correlated with percent base saturation. However, differences between watersheds in soil N substrate caused by presence of leguminous black locust in the disturbed watershed were confounded with differences in soil acidity. Denitrification was most strongly affected by soil moisture, which in turn was determined by precipitation events and slope position. Gaseous N losses from well drained midslope and toeslope landscape positions appeared to be minor relative to other N transformations. Favorable conditions for denitrification occurred at a poorly drained site near the stream of the disturbed watershed. Laboratory incubations revealed high rates of NO(3) reduction in these soils. We speculate that the riparian zone is a major site of depletion of NO(3) from the soil solution via denitrification. PMID- 16347235 TI - Palo Podrido: Model for Extensive Delignification of Wood by Ganoderma applanatum. AB - Chemical and micromorphological analysis revealed that South Chilean "palo podrido" results from a white-rot fungus that causes highly selective and extensive delignification. Palo podrido samples from 10 different hardwood trunks (Eucryphia cordifolia, Drimys winteri, and Nothofagus dombeyi) decayed by Ganoderma applanatum were analyzed. Of 14 samples, 11 had extremely low Klason lignin values, ranging from 6.1 to 0.4% (dry weight). The most remarkable and unusual feature was that delignification and defibration were not restricted to small pockets but extended throughout large areas in the interior of trunks subjected to undisturbed rotting over long periods of time. Comparative analysis of water content, swelling capacity, and lignin content led to the conclusion that besides lignin degradation, suppression of the cellulolytic activity of the rotting organisms plays a decisive role. Among various nutrients added to a palo podrido sample (3% residual Klason lignin), the nitrogen source was the only one leading to almost complete cellulose degradation. We suggest that the extremely low nitrogen content (0.037 to 0.073% [dry weight]) of the investigated wood species was the primary cause for the extensive delignification as well as the concomitant suppression of cellulose breakdown. The low temperatures, high humidity, and microaerobic conditions maintained within the decaying trunks are discussed as additional ecological factors favoring delignification in South Chilean rain forests. PMID- 16347236 TI - Isolation of Halotolerant Thermus spp. from Submarine Hot Springs in Iceland. AB - Thermophilic, aerobic bacteria of the genus Thermus were isolated from submarine alkaline hot springs in Iceland. Five submarine hot springs were sampled, and all had viable counts of Thermus spp. of about 10 CFU/ml. All submarine strains grew in the presence of NaCl at 3% or higher, but no strains from terrestrial hot springs would grow at concentrations higher than 1% NaCl. The growth rate of submarine Thermus strains was not stimulated by NaCl and was reduced at NaCl concentrations higher than 1%. The pattern of growth of these isolates on single carbon sources was similar to that of terrestrial isolates. PMID- 16347237 TI - Production and turnover of planktonic bacteria in two southeastern blackwater rivers. AB - Production by attached and free-living planktonic bacteria in two blackwater rivers in the Southeastern United States was measured over a period of 14 months by using the rate of incorporation of [methyl-H]thymidine into DNA. Production rates and biomass dynamics were compared to determine the potential for in situ production to supply planktonic biomass. Bacterial production in these rivers was moderate and varied seasonally. Rates varied from 0.058 to 2.120 mg of C m h in the Ogeechee River and from 0.002 to 2.418 mg of C m h in Black Creek. Regressions of growth rate on various environmental variables showed that temperature and total dissolved organic carbon concentration were the best predictors of growth. Although attached bacteria were <21% of the total biomass, they accounted for up to 53% of the total production. Turnover times for attached bacteria ranged from <1 day to >3 years depending on season. Turnover times of free-living bacteria varied from 4.4 days to 11.8 years. Comparisons of biomass with production indicated that during most seasons, the majority of bacterial biomass in these rivers was of allochthonous origin. During summer, when water temperatures were high, bacterial growth in the river may have supplied a greater percentage of the standing stock of bacteria than allochthonous inputs. PMID- 16347238 TI - Novel actinomycete isolated from bulking industrial sludge. AB - A novel actinomycete was the predominant filamentous microorganism in bulking activated sludge in a bench-scale reactor treating coke plant wastewater. The bacterium was isolated and identified as an actinomycete that is biochemically and morphologically similar to Amycolatopsis orientalis; however, a lack of DNA homology excludes true relatedness. At present, the isolate (NRRL B 16216) cannot be assigned to the recognized taxa of actinomycetes. PMID- 16347239 TI - Effect of temperature on growth, respiration, and nutrient regeneration by an omnivorous microflagellate. AB - The effect of temperature on the rates and extent of carbon and nitrogen cycling by the heterotrophic microflagellate Paraphysomonas imperforata (diameter, 7 to 12 mum) fed with the diatom Phaeodactylum tricornutum was investigated over an ecologically pertinent temperature range (14 to 26 degrees C). All physiological rates investigated increased with increasing temperature. Q(10) values were similar for all rate changes and were comparable to those which have been reported for other protozoa. In contrast to all rates, microflagellate gross growth efficiency and cell volume were unaffected by temperature. Decreases in the concentrations of particulate carbon and particulate nitrogen from grazed diatom cultures also were similar when summed over the entire growth phase of the microflagellate population. Therefore, the proportions of ingested carbon and nitrogen which were incorporated or remineralized by the microflagellate were independent of temperature between 14 and 26 degrees C. At temperatures above 18 degrees C, growth rates of P. imperforata were greater than the maximum growth rates reported for most phytoplankton. We conclude that the impact of P. imperforata on natural phytoplankton communities is not controlled by temperature above 18 degrees C but may be affected by the rate at which zooplankton or microzooplankton prey on the microflagellate, as well as the inability of the microflagellate to graze efficiently when phytoplankton are present at low cell densities. PMID- 16347240 TI - Cloning of a Gene Cluster from Cellvibrio mixtus which Codes for Cellulase, Chitinase, Amylase, and Pectinase. AB - The soil isolate Cellvibrio mixtus UQM2294 degraded a variety of polysaccharides including microcrystalline cellulose. Among 6,000 cosmid clones carrying C. mixtus DNA, constructed in Escherichia coli with pHC79, 50 expressed the ability to degrade one or more of the following substrates: carboxymethyl cellulose, chitin, pectin (polygalacturonic acid), cellobiose, and starch. These degradative genes are encoded in a single 94.1-kilobase segment of the C. mixtus genome; a preliminary order of the genes is starch hydrolysis, esculin hydrolysis, cellobiose utilization, chitin hydrolysis, carboxymethyl cellulose hydrolysis, and polygalacturonic acid hydrolysis. A restriction endonuclease cleavage map was constructed, and the genes for starch, carboxymethyl cellulose, cellobiose, chitin, and pectin hydrolysis were subcloned. PMID- 16347241 TI - Spatial and Temporal Variations in Bacterial Macromolecule Labeling with [methyl H]Thymidine in a Hypertrophic Lake. AB - The incorporation of [methyl-H]thymidine into three macromolecular fractions, designated as DNA, RNA, and protein, by bacteria from Hartbeespoort Dam, South Africa, was measured over 1 year by acid-base hydrolysis procedures. Samples were collected at 10 m, which was at least 5 m beneath the euphotic zone. On four occasions, samples were concurrently collected at the surface. Approximately 80% of the label was incorporated into bacterial DNA in surface samples. At 10 m, total incorporation of label into bacterial macromolecules was correlated to bacterial utilization of glucose (r = 0.913, n = 13, P < 0.001). The labeling of DNA, which ranged between 0 and 78% of total macromolecule incorporation, was inversely related to glucose uptake (r = -0.823), total thymidine incorporation (r = -0.737), and euphotic zone algal production (r = -0.732, n = 13, P < 0.005). With decreased DNA labeling, increasing proportions of label were found in the RNA fraction and proteins. Enzymatic digestion followed by chromatographic separation of macromolecule fragments indicated that DNA and proteins were labeled while RNA was not. The RNA fraction may represent labeled lipids or other macromolecules or both. The data demonstrated a close coupling between phytoplankton production and heterotrophic bacterial activity in this hypertrophic lake but also confirmed the need for the routine extraction and purification of DNA during [methyl-H]thymidine studies of aquatic bacterial production. PMID- 16347242 TI - Ingestion and Adsorption of Bacillus thuringiensis subsp. israelensis by Gammarus lacustris in the Laboratory. AB - Several groups of Gammarus lacustris adults were exposed to solutions containing 0.5 and 5.0 mg of Bacillus thuringiensis subsp. israelensis per liter for 1- or 24-h periods by using traditional static bioassay exposure procedures. During a postexposure holding period, fecal pellets were removed and plated on tryptic soy agar to determine B. thuringiensis subsp. israelensis spore content. The experiments verified that traditional exposure procedures assure ingestion of B. thuringiensis subsp. israelensis spores and provided a mean dose estimate of 1,948 spores ingested per test animal with a 95% confidence interval ranging from 891 to 4,296 (1-h exposure, 5.0 mg/liter). It was also found that dose level is highly dependent upon both exposure duration and concentration and that relatively short exposures can result in a relatively long-term retention of spores postexposure (>/=30 days). Body burden experiments established that large numbers of spores adsorb to the bodies of test animals during exposure and may in part explain the long-term retention of spores in the test system postexposure. These results imply that in field applications of microbial control agents, toxicologically unaffected but exposed organisms might transport the agent to untreated sites, expanding the effective treatment area and the number of organisms exposed. PMID- 16347243 TI - Bacterial enhancement of vinyl fouling by algae. AB - The role of bacteria in the development of algae on low-density vinyl was investigated. Unidentified bacterial contaminants in unialgal stock cultures of Phormidium faveolarum and Pleurochloris pyrenoidosa enhanced, by 1 to 2 orders of magnitude, colonization of vinyl by these algae, as determined by epifluorescence microscopy counts and chlorophyll a in extracts of colonized vinyl. Colonization by bacteria always preceded that by algae. Scanning electron microscopy of the colonized Phormidium-bacteria mixture revealed the presence of a slime matrix engulfing both bacteria and algae that may have facilitated algal attachment. Slime was not evident in the Pleurochloris-bacteria mixture, suggesting that the attachment mechanisms differ for the two algae. PMID- 16347244 TI - Microbial oxidation of hydrocarbons and related compounds by whole-cell suspensions of the methane-oxidizing bacterium h-2. AB - Previously, a thermophilic obligate methane-oxidizing bacterium, H-2 (type I), was isolated in our laboratory. H-2 is a new type of methylotroph because of the G+C content of DNA; it uses both the ribulose monophosphate pathway and the serine pathway for carbon assimilation and possesses a new quinone. In addition, we found that resting cell suspensions of H-2 had the ability to oxidize a variety of compounds different from the other methane-oxidizing bacteria as follows. (i) C(1) to C(8)n-alkanes are hydroxylated and further oxidized, yielding mixtures of the corresponding alcohols, aldehydes, acids, and ketones. Liquid alkanes are transformed through a different oxidative pathway from that of gaseous ones. (ii) Both gaseous (C(2) to C(4)) and liquid (C(5), C(6)) n-alkenes are oxidized to their corresponding 1,2-epoxides. (iii) Liquid monochloro and dichloro n-alkanes (C(5), C(6)) are oxidized, yielding their corresponding acids or haloacids. (iv) Diethyl ether is oxidized to acetic acid; no ethanol and acetaldehyde are detected. (v) Cyclic and aromatic compounds are also oxidized. (vi) Secondary alcohols (C(3) to C(10)) are oxidized to their corresponding methyl ketones. PMID- 16347245 TI - Mutagenesis of Protoplasts and Regeneration of Mycelium in the Mushroom Volvariella volvacea. AB - Regenerating protoplasts were obtained from mycelial culture of the mushroom Volvariella volvacea by the action of the lytic enzyme Novozym 234 in the presence of 0.01 M phosphate buffer (pH 6.0) containing 0.6 M NaCl. Regeneration was found to be poor in liquid medium, but more than 50% regeneration was achieved on solid 2% agar medium overlaid with 0.5% agar. Protoplasts of V. volvacea were found to be highly sensitive to the killing action of both UV irradiation and N-methyl-N'-nitro-N-nitrosoguanidine. However, no morphological or auxotrophic mutants could be obtained from protoplasts by chemical mutagenesis. Four types of morphological mutants and one auxotrophic (adenine negative) mutant were obtained from UV-irradiated protoplasts. The adenine negative mutant of V. volvacea was found to be stable, not losing auxotrophy on repeated subculture. PMID- 16347246 TI - End products of anaerobic chitin degradation by salt marsh bacteria as substrates for dissimilatory sulfate reduction and methanogenesis. AB - The anaerobic pathway of chitin decomposition by chitinoclastic bacteria was examined with an emphasis on end product coupling to other salt marsh bacteria. Actively growing chitinoclastic bacterial isolates produced primarily acetate, H(2), and CO(2) in broth culture. No sulfate-reducing or methanogenic isolates grew on chitin as sole carbon source or produced any measurable degradation products. Mixed cultures of chitin degraders with sulfate reducers resulted in positive sulfide production. Mixed cultures of chitin-degrading isolates with methanogens resulted in the production of CH(4) with reductions in headspace CO(2) and H(2). The combination of all three metabolic types resulted in the simultaneous production of methane and sulfide, with more methane being produced in mixed cultures containing CO(2)-reducing methanogens and acetoclastic sulfate reducers because of less interspecific H(2) competition. PMID- 16347247 TI - Changes in Protein Composition of Three Bacterial Isolates from Marine Waters during Short Periods of Energy and Nutrient Deprivation. AB - Two-dimensional gel electrophoresis analysis of and total cell protein determination for three bacterial isolates from marine waters at the onset and after 24 h of energy and nutrient deprivation demonstrated that the three isolates exhibited different pathways of starvation survival. Two strains appeared to synthesize new proteins during starvation. PMID- 16347248 TI - Symbiotic Characteristics and Rhizobium Requirements of a Leucaena leucocephala x Leucaena diversifolia Hybrid and Its Parental Genotypes. AB - In 56-day-old plants, Leucaena leucocephala and its hybrid with L. diversifolia showed 100% more total N than did L. diversifolia. Significant (P < 0.01) host inoculation interaction in total N was 14.4% of the total phenotypic variation. The most effective and competitive Rhizobium sp. for the leucaenas was TAL 1145. Three-strain mixed inoculation was inferior to TAL 1145 alone. PMID- 16347249 TI - Effects of genotype, spawn run time, and substrate formulation on biological efficiency of shiitake. AB - Substrate formulations of sawdust, wheat bran, and millet were inoculated with spawns from one hybrid and two parental genotypes of Lentinula edodes. Biological efficiency (BE) and size data on mushrooms harvested from two substrate formulations with spawn run times of 60, 90, and 120 days were analyzed. A significant genotype-spawn run time-substrate formulation interaction was observed for BE. The longer spawn runs resulted in greater BE than the shorter spawn runs. This study points to the need for more work to determine the underlying variability within genotypes. PMID- 16347250 TI - Comparative growth of natural bacterial isolates on various lignin-related compounds. AB - Bacterial strains were isolated on the basis of their ability to proliferate in a minimal medium containing one of a series of lignin-related compounds as the sole carbon and energy source. These included the aromatic monomers guaiacol, vanillic and coumaric acids, a dimer and a trimer possessing the arylglycerol-beta-aryl ether linkage, anisoin, and both the ether-soluble and -insoluble fractions of kraft lignin. The growth of the strains on each of these compounds was measured. The results showed that the metabolic properties of the strains varied according to the structure of the carbon sources used for their selection. Spectrophotometric tracings of the culture medium during the log phase of growth of one of the strains on the beta-O-4 dimer revealed decomposition with the release of guaiacol. PMID- 16347251 TI - Methane Production from Formate by Syntrophic Association of Methanobacterium bryantii and Desulfovibrio vulgaris JJ. AB - Coculture of a sulfate-reducing bacterium, when grown in the absence of added sulfate, with Methanobacterium bryantii, which uses only H(2) and CO(2) for methanogenesis, degraded formate to CH(4). A pure culture of Desulfovibrio vulgaris JJ was able to produce small amounts of H(2). Such a syntrophic relationship might provide an additional way to avoid formate accumulation in anaerobic environments. PMID- 16347252 TI - Effects of Monovalent and Divalent Salts on the Phospholipid and Fatty Acid Compositions of a Halotolerant Planococcus sp. AB - [This corrects the article on p. 581 in vol. 52.]. PMID- 16347253 TI - Validity of eucaryote inhibitors for assessing production and grazing mortality of marine bacterioplankton. AB - Application of eucaryote inhibitors to the estimation of production and grazing mortality of bacterioplankton was evaluated. Exposure to a range of concentrations of thiram, cycloheximide, and neutral red (0.4 to 210, 36 to 1,777, 4 to 346 muM, respectively) was 98 to 100% effective at inhibiting growth of a chrysomonad in culture. Exposure to colchicine and griseofulvin (50 to 1,000 muM for both) yielded only 24 to 94 and 53 to 79% inhibition, respectively. Exposures to thiram, neutral red, and griseofulvin were 90 to 100% effective at inhibiting growth in culture of a ciliate, Cyclidium sp., and the responses to colchicine and cycloheximide were variable (64 to 100 and 0 to 100% inhibition, respectively). Thiram and neutral red inhibited field populations of nanozooplankton more effectively than cycloheximide and colchicine. Direct effects of eucaryote inhibitors on growing cultures of bacterioplankton varied with parameters measured and duration of exposure. After 3-day exposures, specific growth rates and "instantaneous" heterotrophic potential ([C]glucose uptake) were not consistently affected, but biosynthetic activity (RNA and DNA syntheses) was depressed. The degree of inhibition of isolates and field populations of phytoplankton depended upon type of inhibitor and phytoplankton species. In field experiments, it was possible to calculate rates of bacterioplankton production and grazing mortality for only 16 of 29 inhibitor experiments and for 4 of 10 size fractionation experiments. Bacterioplankton production and mortality estimates varied greatly with the eucaryote inhibitor used, and those derived from inhibition techniques were substantially different from those derived from fractionation techniques. The poor performances of both techniques are attributed to the following: (i) effects of inhibitors on phytoplankton, (ii) indirect effects of the inhibitors on bacterioplankton, and (iii) insufficient separation of grazers from prey by filtration techniques. Because of the inconsistent results obtained in this investigation, we strongly recommend exercising caution in the application of inhibitor techniques to ecological problems, especially in phototrophically dominated systems. PMID- 16347254 TI - Microbiological Degradation of Malodorous Substances of Swine Waste under Aerobic Conditions. AB - Phenol, p-cresol, and volatile fatty acids (VFA; acetic, propionic, isobutyric, butyric, isovaleric, and valeric acids) were used as odor indicators of swine waste. Aeration of the waste allowed the indigenous microorganisms to grow and degrade these malodorous substances. The time required for degradation of these substances varied according to the waste used, and it was not necessarily related to their concentrations. Using a minimal medium which contained one of the malodorous compounds as sole carbon source, we have selected from swine waste microorganisms that can grow in the medium. The majority of these microorganisms were able to degrade the same substrate when inoculated in sterilized swine waste but with an efficiency varying from one strain to the other. None of these strains was able to degrade all malodorous substances studied. Within 6 days of incubation these selected strains degraded the following: Acinetobacter calcoaceticus, phenol and all VFA; Alcaligenes faecalis, p-cresol and all VFA; Corynebacterium glutamicum and Micrococcus sp., phenol, p-cresol, and acetic and propionic acids; Arthrobacter flavescens, all VFA. On a laboratory scale, the massive inoculation of swine waste with C. glutamicum or Micrococcus sp. accelerated degradation of the malodorous substances. However, this effect was not observed with all of the various swine wastes tested. These results suggest that an efficient deodorization process of various swine wastes could be developed at the farm level based on the aerobic indigenous microflora of each waste. PMID- 16347255 TI - Cricket Paralysis Virus, a Potential Control Agent for the Olive Fruit Fly, Dacus oleae Gmel. AB - Representatives of several families of insect viruses were tested for growth and pathogenicity in the olive fruit fly, Dacus oleae Gmel. The viruses included nuclear polyhedrosis viruses, an iridovirus, two picornaviruses, and Trichoplusia ni small RNA virus (a member of the Nudaurelia beta family), in addition to two naturally occurring viruses of the olive fruit fly. Two viruses, one of the two picornaviruses (cricket paralysis virus [CrPV] and the iridovirus (type 21 from Heliothis armigera), were found to replicate in adult flies. Flies which were fed on a solution containing CrPV for 1 day demonstrated a high mortality with 50% dying within 5 days and nearly 80% dying within 12 days of being fed. The virus was transmissible from infected to noninfected flies by fecal contamination. The CrPV which replicated in the infected flies was demonstrated to be the same as input virus by infection of Drosophila melanogaster cells and examination of the expressed viral proteins, immunoprecipitation of the virus purified from flies, and electrophoretic analysis of the structural proteins. PMID- 16347256 TI - Detection of Specific Strains and Variants of Streptococcus cremoris in Mixed Cultures by Immunofluorescence. AB - Antisera against four different strains of Streptococcus cremoris were raised by injecting rabbits with washed suspensions of whole cells. These antisera interacted specifically with the corresponding strain in a mixture of up to nine different S. cremoris strains. The antisera could be used for analyzing the composition of mixed cultures containing these strains by immunofluorescence. Competition experiments were performed in batch and continuous cultures under amino acid limitation. A bacteriophage-sensitive variant of S. cremoris SK11 (SK1128) could be distinguished from a bacteriophage-resistant variant (SK1143) by the same immunofluorescence technique. The competition between the two variants and the stability of both variants in pure cultures were followed with the specific antibodies. Antibodies against the purified proteolytic system of S. cremoris Wg2 were used to determine the presence of proteases by immunofluorescence in several S. cremoris strains under different culture conditions. The described immunofluorescence methods can be used to analyze complex mixed starter cultures common in the dairy industry as the strains and variants present in these mixtures can be recognized microscopically. PMID- 16347257 TI - Root-Zone-Specific Oxygen Tolerance of Azospirillum spp. and Diazotrophic Rods Closely Associated with Kallar Grass. AB - The effect of oxygen on N(2)-dependent growth of two Azospirillum strains and two diazotrophic rods closely associated with roots of Kallar grass (Leptochloa fusca) was studied. To enable precise comparison, bacteria were grown in dissolved-oxygen-controlled batch and continuous cultures. Steady states were obtained from about 1 to 30 muM O(2), some of them being carbon limited. All strains needed a minimum amount of oxygen for N(2)-dependent growth. Nitrogen contents between 10 and 13% of cell dry weight were observed. The response of steady-state cultures to increasing O(2) concentrations suggested that carbon limitation shifted to internal nitrogen limitation when N(2) fixation became so low that the bacteria could no longer meet their requirements for fixed nitrogen. For Azospirillum lipoferum Rp5, increase of the dilution rate resulted in decreased N(2) fixation in steady-state cultures with internal nitrogen limitation. Oxygen tolerance was found to be strain specific in A. lipoferum with strain Sp59b as a reference organism. Oxygen tolerance of strains from Kallar grass was found to be root zone specific. A. halopraeferens Au 4 and A. lipoferum Rp5, predominating on the rhizoplane of Kallar grass, and strains H6a2 and BH72, predominating in the endorhizosphere, differed in their oxygen tolerance profiles. Strains H6a2 and BH72 still grew and fixed nitrogen in steady-state cultures at O(2) concentrations exceeding those which absolutely inhibited nitrogen fixation of both Azospirillum strains. It is proposed that root-zone specific oxygen tolerance reflects an adaptation of the isolates to the microenvironments provided by the host plant. PMID- 16347258 TI - Filter Paper Dot-Immunobinding Assay for Detection of Spiroplasma citri. AB - A rapid filter paper dot-immunobinding assay was adapted to detect the wall-less mollicute Spiroplasma citri in medium, plants, or insects. Filter paper spotted with sample was incubated first in dilute antiserum, then in protein A peroxidase, and finally in a substrate of 4-chloro-1-naphthol plus hydrogen peroxide. The detection limit averaged 2.3 x 10 CFU/ml in cultures, and S. citri was detected in single infected leafhoppers. This assay was less sensitive but more rapid and economical than an enzyme-linked immunosorbent assay. PMID- 16347260 TI - Close association of azospirillum and diazotrophic rods with different root zones of kallar grass. AB - [This corrects the article on p. 522 in vol. 52.]. PMID- 16347259 TI - Nitrosubstituted aromatic compounds as nitrogen source for bacteria. AB - Bacteria which utilized nitroaromatic compounds (0.5 mM) as sole source of nitrogen were isolated from soil. With 2,6-dinitrophenol and succinate as carbon source, a Pseudomonas strain was isolated which liberated and assimilated nitrite. Approximately 2 mol of NO(2) per mol of 2,6-dinitrophenol was released by resting cells. The xenobiotic compound was totally degraded, although specific growth yields were low even with succinate as a carbon source. PMID- 16347261 TI - Phenotypic characteristics of coliform and noncoliform bacteria from a public water supply compared with regional and national clinical species. AB - [This corrects the article on p. 476 in vol. 52.]. PMID- 16347262 TI - Lipoteichoic acids in lactobacillus strains that colonize the mouse gastric epithelium. AB - [This corrects the article on p. 302 in vol. 52.]. PMID- 16347263 TI - Biofilm Dynamics and Kinetics during High-Rate Sulfate Reduction under Anaerobic Conditions. AB - The sulfate kinetics in an anaerobic, sulfate-reducing biofilm were investigated with an annular biofilm reactor. Biofilm growth, sulfide production, and kinetic constants (K(m) and V(max)) for the bacterial sulfate uptake within the biofilm were determined. These parameters were used to model the biofilm kinetics, and the experimental results were in good agreement with the model predictions. Typical zero-order volume rate constants for sulfate reduction in a biofilm without substrate limitation ranged from 56 to 93 mumol of SO(4)-cm h at 20 degrees C. The temperature dependence (Q(10)) of sulfate reduction was equivalent to 3.4 at between 9 and 20 degrees C. The measured rates of sulfate reduction could explain the relatively high sulfide levels found in sewers and wastewater treatment systems. Furthermore, it has been shown that sulfate reduction in biofilms just a few hundred micrometers thick is limited by sulfate diffusion into biofilm at concentrations below 0.5 mM. This observation might, in some cases, be an explanation for the relatively poor capacity of the sulfate-reducing bacteria to compete with the methanogenic bacteria in anaerobic wastewater treatment in submerged filters. PMID- 16347264 TI - Rhizobial Ecology of the Woody Legume Mesquite (Prosopis glandulosa) in the Sonoran Desert. AB - Soil samples were collected from the surface (0 to 0.6 m) and phreatic (3.9 to 4.5 m) root systems of a Prosopis glandulosa woodland in the Sonoran Desert of southern California. P. glandulosa seedlings were inoculated with these soils, and rhizobia were isolated from nodules. The phreatic soil, characterized by constant moisture and temperature but low nutrient availability, favored slow growing (SG) isolates as nodule occupants (85%). SG isolates from the surface and phreatic soil were distinct based on differences in colony morphology. Isolates from the surface soil, characterized by high nutrient availability and widely fluctuating water content and temperature, were equally represented by fast growing and SG rhizobia. Most SG isolates (83%) had nodule relative efficiencies of <0.80, whereas 54% of the fast-growing isolates had relative efficiency values of >0.80. PMID- 16347265 TI - Variation in Phospholipid Ester-Linked Fatty Acids and Carotenoids of Desiccated Nostoc commune (Cyanobacteria) from Different Geographic Locations. AB - Profiles of phospholipid fatty acids and carotenoids in desiccated Nostoc commune (cyanobacteria) collected from China, Federal Republic of Germany, and Antarctica and in axenic cultures of the desiccation-tolerant strains N. commune UTEX 584 and Hydrocoleum strain GOEI were analyzed. The phospholipid fatty acid contents of the three samples of desiccated Nostoc species were all similar, and the dominant compounds were 16:1omega7c, 16:0, 18:2omega6, 18:3omega3, and 18:1omega7c. In comparison with the field materials, N. commune UTEX 584 had a much higher ratio of 18:2omega6 to 18:3omega3 (5.36) and a significantly lower ratio of 18:1omega7c to 18:1omega9c (1.86). Compound 18:3 was present in large amounts in the samples of desiccated Nostoc species which had been subject, in situ, to repeated cycles of drying and rewetting, but represented only a small fraction of the total fatty acids of the strains grown in liquid culture. This finding is in contrast to the data obtained from studies on the effects of drought and water stress on higher plants. Field materials of Nostoc species contained, in contrast to the axenic strains, significant amounts of apocarotenoids and a P384 pigment which, upon reduction with NaBH(4), yielded a mixture of a chlorophyll derivative and a compound with an absorption maximum of 451 nm. A clear distinction can be made between the carotenoid contents of the axenic cultures and the desiccated field materials. In the former, beta-carotene and echinenone predominate; in the latter, canthaxanthin and the beta-gamma series of carotenoids are found. PMID- 16347266 TI - Formation and regeneration of protoplasts of the actinorhizal nitrogen-fixing actinomycete frankia. AB - Procedures for forming and regenerating protoplasts of four Frankia strains are described. Cells obtained from growth medium containing 0.1% glycine were digested with lysozyme (250 mug/ml) in a medium containing 0.5 M sucrose, 5.0 mM CaCl(2), and 5.0 mM MgCl(2). Protoplasts were formed during 15 to 120 min of digestion at 25 degrees C. Optimum conditions for protoplast regeneration involved placing protoplasts on a layer of complex growth medium containing 0.3 M sucrose, 5.0 mM CaCl(2), and 5.0 mM MgCl(2) which was overlaid with a layer of 0.8% low-melting-point agarose containing 0.5 M sucrose, 5.0 mM MgCl(2), and 5.0 mM CaCl(2). The maximum regeneration efficiency was 36.9% for strain CpI1, 1.3% for strain ACN1, 27% for strain EAN1pec, and 20% for strain EuI1c. PMID- 16347267 TI - Role of Carbon Dioxide in Catabolism of Propane by "Nocardia paraffinicum" (Rhodococcus rhodochrous). AB - The catabolism of propane by "Nocardia paraffinicum" (Rhodococcus rhodochrous) has been shown to involve CO(2) fixation after its oxidation to propionic acid. "N. paraffinicum" failed to grow on either propane or 1-propanol in the absence of CO(2). The rate of propane utilization was directly related to the initial CO(2) concentration, and Warburg respirometry suggested that CO(2) was required for the catabolism of 1-propanol, propionaldehyde, and propionate but not for 2 propanol. These data also suggested that the predominant pathway for the utilization of propane by "N. paraffinicum" was through 1-propanol. The use of [2 C]propane and CO(2) confirmed the catabolism of propane and the fixation of CO(2). Through the use of these isotopes and the pyruvate carboxylase inhibitor sodium arsenite, the labeled 2,4-dinitrophenylhydrazine derivative of pyruvate was trapped and isolated via thin-layer chromatography. The trapping of [C]pyruvate in this manner was considered to be indicative of the presence of the methylmalonyl coenzyme A pathway for CO(2) fixation. PMID- 16347268 TI - Development of High-Frequency Delivery System for Transposon Tn919 in Lactic Streptococci: Random Insertion in Streptococcus lactis subsp. diacetylactis 18 16. AB - The conjugative transposon Tn919, originally isolated in Streptococcus sanguis FC1, is capable of low-frequency transfer (10 and 10 per recipient) on membrane filters to a wide number of streptococcal recipients including the industrially important lactic streptococci. The introduction of pMG600 (Lac Lax; a lactose plasmid capable of conjugative transfer at high frequencies and which, in certain hosts, confers an unusual clumping phenotype) into a Streptococcus lactis CH919 donor, generating S. lactis CH001, resulted in a significant improvement in the transfer frequency of Tn919 to S. lactis CK50 (1.25 x 10 per recipient). In addition, these matings could be performed on agar surfaces, allowing the recovery of a greater number of recipients than with filter matings. Tn919 also transferred at high frequency to S. lactis subsp. diacetylactis 18-16S but not to Streptococcus cremoris strains. Insertion in 18-16S transconjugants generated from filter matings with an S. lactis CH919 donor was random, occurring at different sites on the chromosome and also in plasmid DNA. Thus, the conditions necessary for the practical exploitation of Tn919 in the targeting and cloning of genes from a member of the lactic streptococci, namely, high-frequency delivery and random insertion in host DNA, were achieved. PMID- 16347269 TI - Effects of Hydrogen Pressure during Growth and Effects of Pregrowth with Hydrogen on Acetate Degradation by Methanosarcina Species. AB - Methanosarcina barkeri 227 and Methanosarcina mazei S-6 grew with acetate as the substrate; we found little effect of H(2) on the rate of aceticlastic growth in the presence of various H(2) pressures between 2 and 810 Pa. We used physical (H(2) addition or flushing the headspace to remove H(2)) and biological (H(2) producing or -utilizing bacteria in cocultures) methods for controlling H(2) pressure in Methanosarcina cultures growing on acetate. Added H(2) (ca. 100 Pa) was removed rapidly (a few hours) by M. barkeri and slowly (within a day) by M. mazei. When the H(2) produced by the aceticlastic methanogens was removed by coculturing with an H(2)-using Desulfovibrio sp., the H(2) pressure was about 2.2 Pa. Under these conditions the stoichiometry of aceticlastic methanogenesis did not change. H(2)-grown inocula of M. barkeri grew with acetate as the sole catabolic substrate if the inoculum culture was transferred during logarithmic growth to acetate-containing medium or if the transfer was accomplished within 1 or 2 days after exhaustion of H(2). H(2)-grown cultures incubated for 4 or more days after exhaustion of H(2) were able to grow with H(2) but not with acetate as the sole catabolic substrate. Addition of small quantities of H(2) to acetate containing medium permitted these cultures to initiate growth on acetate. PMID- 16347270 TI - Estimation of bacterial nitrate reduction rates at in situ concentrations in freshwater sediments. AB - A method was developed to follow bacterial nitrate reduction in freshwater sediments by using common high-performance liquid chromatographic equipment. The low detection limit (14 pmol) of the method enabled us to study concentration profiles and reaction kinetics under natural conditions. Significant nitrate concentrations (1 to 27 muM) were observed in the sediment of Lake Vechten during the nonstratified period; the concentration profiles showed a successive depletion of oxygen, nitrate, and sulfate with depth. The profiles were restricted to the upper 3 cm of the sediment which is rich in organics and loosely structured. Nitrate reduction in the sediment-water interface followed first-order reaction kinetics at in situ concentrations. Remarkably high potential nitrate-reducing activity was observed in the part of the sediment in which nitrate did not diffuse. This activity was also observed throughout the whole year. Estimates of K(m) varied between 17 and 100 muM and V(max) varied between 7.2 and 36 mumol cm day for samples taken at different depths. The diffusion coefficient of nitrate ([10 +/- 0.4] x 10 cm s) across the sediment water interface was estimated by a constant-source technique and applied to a mathematical model to estimate the net nitrate reduction during the nonstratified period. In this period, observed nitrate reduction rates by the model, 0.2 to 0.4 mmol m day, were lower than those found for oxygen (27 mmol m day) and sulfate (0.4 mmol m day). During the summer stratification, nitrate was absent in the sediment and reduction could not be estimated by the model. PMID- 16347271 TI - Surface-active agents from two bacillus species. AB - Two Bacillus species were studied which produced bioemulsifiers; however, they were distinctly different compounds. Bacillus sp. strain IAF 343 produced unusually high yields of extracellular biosurfactant when grown on a medium containing only water-soluble substrates. The yield of 1 g/liter was appreciably better than those of most of the biosurfactants reported previously. This neutral lipid product, unlike most lipid biosurfactants, had significant emulsifying properties. It did not appreciably lower the surface tension of water. On the same medium, Bacillus cereus IAF 346 produced a more conventional polysaccharide bioemulsifier, but it also produced a monoglyceride biosurfactant. The bioemulsifier contained substantial amounts of glucosamine and originated as part of the capsule layer. The monoglyceride lowered the surface tension of water to 28 mN/m. It formed a strong association with the polysaccharide, and it was necessary to use ultrafiltration to effect complete separation. The removal of the monoglyceride caused the polysaccharide to precipitate. It is suggested that earlier reports of biopolymers which both stabilized emulsions and lowered surface tension were actually similar aggregates of lipid and bioemulsifier. PMID- 16347272 TI - Expression Vector for Zymomonas mobilis. AB - This study describes the construction of several useful cloning vectors which can be conjugated from Escherichia coli into Zymomonas mobilis at high frequency, approaching 10 per donor or recipient. These vectors contain a broad-host-range replicon and mob site from RSF1010, a chloramphenicol acyltransferase gene under the control of an enteric consensus promoter, and a second mob site (originally derived from RP4). The addition of this second mob site appears to be responsible for a 2-order-of-magnitude increase in the efficiency of transfer into Z. mobilis. Such vectors may be useful for other gram-negative bacteria in which conjugation efficiencies are low. These vectors are stably maintained in Z. mobilis with no detectable loss of plasmid after 50 generations in the absence of selective pressure. One of these, pLOI193, contains the tetracycline gene from pBR322 and associated cloning sites for insertional inactivation. Another, pLOI204, contains a Z. mobilis promoter immediately upstream from a BamHI site which can be used for cloning. This promoter has been shown to efficiently drive the expression of beta-galactosidase in both Z. mobilis and E. coli. This promoter fragment from Z. mobilis has been sequenced, and the site for transcriptional initiation in E. coli and Z. mobilis has been identified. PMID- 16347273 TI - Selection and improvement of lignin-degrading microorganisms: potential strategy based on lignin model-amino Acid adducts. AB - The purpose of this investigation was to test a potential strategy for the ligninase-dependent selection of lignin-degrading microorganisms. The strategy involves covalently bonding amino acids to lignin model compounds in such a way that ligninase-catalyzed cleavage of the models releases the amino acids for growth nitrogen. Here we describe the synthesis of glycine-N-2-(3,4 dimethoxyphenyl)ethane-2-ol (I) and demonstrate that growth (as measured by mycelial nitrogen content) of the known lignin-degrading basidiomycete Phanerochaete chrysosporium Burds. with compound I as the nitrogen source depends on its production of ligninase. Ligninase is shown to catalyze the oxidative C-C cleavage of compound I, releasing glycine, formaldehyde, and veratraldehyde at a 1:1:1 stoichiometry. P. chrysosporium utilizes compound I as a nitrogen source, but only after the cultures enter secondary metabolism (day 3 of growth), at which time the ligninase and the other components of the ligninolytic system (lignin --> CO(2)) are expressed. Compound I and related adducts have potential not only in the isolation of lignin-degrading microbes but, perhaps of equal importance, in strain improvement. PMID- 16347274 TI - Effect of salinity on mercury-methylating activity of sulfate-reducing bacteria in estuarine sediments. AB - The biomethylation of mercury was measured in anoxic estuarine sediments that ranged in salinity from 0.03 to 2.4% with or without added molybdate, an inhibitor of sulfate reducers. Mercury methylation was inhibited by molybdate by more than 95%, regardless of sediment salinity. In the absence of inhibitor, high salinity sediments methylated mercury at only 40% of the level observed in low salinity sediments. In response to molybdate inhibition of sulfate reducers, methanogenesis increased up to 258% in high-salinity sediments but only up to 25% in low-salinity sediments. In contrast to an earlier low-salinity isolate, a Desulfovibrio desulfuricans strain from high-salinity sediment required 0.5 M sodium for optimal growth and mercury methylation activity. The formation of negatively charged mercuric chloride complexes at high salinity did not noticeably interfere with the methylation process. Results of these studies demonstrate that sulfate reducers are responsible for mercury methylation in anoxic estuarine sediments, regardless of the prevailing salinity. PMID- 16347275 TI - Particle counter determination of bacterial biomass in seawater. AB - The applicability of the Elzone particle counter to the determination of marine bacterial biomass was investigated. The biomass of bacterial pure cultures and a mixed natural population were followed by using the particle counter, a CHN analyzer, and an ATP analyzer. The particle counter showed the precise size distribution of number and volume of submicron-size particles in seawater. For the pure cultured bacterial strains, the conversion factor from volume to carbon is 0.209 mg of C per mm, and for natural bacterial cells of >0.6 mum in diameter, it is 0.184 mg of C per mm. It is recommended that 0.2 be used as the conversion factor for both pure cultured marine bacterial cells and natural bacteria from coastal and near-shore marine environments. PMID- 16347276 TI - Evaluation of slime-producing bacteria in oil field core flood experiments. AB - Epifluorescence microscopy and carbohydrate determinations indicated that the decrease in permeability of oil reservoir sand to reclaimed sewage water was partially the result of biological plugging. Filtration and biocide addition studies demonstrated that the increase in bacterial densities and slime concentrations in flooded oil field cores appeared to be due to both deposition from the reclaimed water and in situ microbial growth and slime production. Although these biological components increased throughout the cores during flooding, the region where the water entered the core exhibited the highest cell densities and slime concentrations. The approach described in this report should be useful in predicting the potential of a water source to induce biological plugging of oil reservoir sand. PMID- 16347277 TI - Production of Microbial Biomass Protein from Potato Processing Wastes by Cephalosporium eichhorniae. AB - The use of Cephalosporium eichhorniae 152 (ATCC 38255) (reclassified as Acremonium alabamense; see Addendum in Proof), a thermophilic, acidophilic, amylolytic fungus, for the conversion of potato processing wastes into microbial protein for use as animal feed was studied. The fungus was not inhibited by alpha solanine or beta-2-chaconine, antimicrobial compounds in potatoes, or by morpholine or cyclohexylamine (additives to steam used in the peeling process) at levels likely to be encountered in this substrate. Mixed effluent from holding tanks at a potato-processing plant contained about 10 bacteria per ml and inhibited fungal growth. The fungus grew well on fresh potato wastes containing up to 5% total carbohydrate and utilized both starch and protein at 45 degrees C and pH 3.75. On potato homogenate medium containing 2% carbohydrate (about 14% fresh potato) supplemented with monoammonium phosphate (0.506 g/liter) and ferric iron (0.1 g/liter), with pH control (at 3.75) and additional nitrogen supplied by the automatic addition of ammonium hydroxide, typical yields were 0.61 g (dry weight) of product and 0.3 g of crude protein per g of carbohydrate supplied. An aerobic, spore-forming bacterium, related to Bacillus brevis, commonly contaminated nonsterilized batch cultures but was destroyed by heating for 15 min at 100 degrees C. PMID- 16347278 TI - Quantification and removal of some contaminating gases from acetylene used to study gas-utilizing enzymes and microorganisms. AB - Acetylene generated from various grades of calcium carbide and obtained from commercial- and purified-grade acetylene cylinders was shown to contain high concentrations of various contaminants. Dependent on the source of acetylene, these included, at maximal values, H(2) (0.023%), O(2) (0.779%), N(2) (3.78%), PH(3) (0.06%), CH(4) (0.073%), and acetone (1 to 10%). The concentration of the contaminants in cylinder acetylene was highly dependent on the extent of cylinder discharge. Several conventional methods used to partially purify cylinder acetylene were compared. A small-scale method for extensively purifying acetylene is described. An effect of acetylene quality on acetylene reduction assays conducted with purified nitrogenase from Azotobacter vinelandii was demonstrated. PMID- 16347279 TI - Factors Involved in Hydrolysis of Microcrystalline Cellulose by Acetivibrio cellulolyticus. AB - Acetivibrio cellulolyticus cellulase obtained by the water elution of residual cellulose from the growth medium was compared with the cellulase activity present in culture supernatants. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that water elution released most of the protein bands which adhered to undigested cellulose from the culture medium. The enzyme in the culture supernatant and that eluted from residual cellulose had specific activities for Avicel hydrolysis that were 20- to 40-fold greater than that of Trichoderma reesei cellulase. However, Ca and a reducing agent such as dithiothreitol were required for maximum Avicel hydrolysis rates by these A. cellulolyticus enzyme preparations. The effect of these agents on p-nitrophenyl lactopyranoside hydrolysis suggested that they were required by an exoglucanase component. Supernatant enzyme preparations contained large amounts of carbohydrate which was separated from most of the cellulase protein by phenyl Sepharose chromatography. Removal of this carbohydrate, which interfered with protein fractionations, allowed for an activity stain analysis of the supernatant enzyme. PMID- 16347280 TI - Selection of Protease-Positive and Protease-Negative Variants of Streptococcus cremoris. AB - Protease-negative variants were shown to outcompete the wild-type strains of Streptococcus cremoris E(8), HP, and Wg(2) at pH values higher than 6.0 in milk. For S. cremoris E(8) this process was studied in more detail. At lower pH values the wild type had a selective advantage. This pH-dependent selection was not found in all media tested. The poor growth of the protease-negative variant at low pH was not due to lower internal pH values. By growing S. cremoris E(8) and Wg(2) in acidified milk (pH 5.9) the proteolytic activity of the cultures could be stabilized. In continuous cultures under amino acid limitation the wild type S. cremoris E(8) and HP strains had a selective advantage over the protease negative variants at low dilution rates (D < 0.2) at all pH values of the medium. This was apparently due to a lower affinity-constant (K(s)) of the protease positive variants for amino acids. Finally, a high fraction of protease-positive variants could be maintained in continuous cultures by using a growth medium with low concentrations of casein as a nitrogen source. At high dilution rates nearly all cells were protease positive. PMID- 16347281 TI - Long-Term Effects of Metal-Rich Sewage Sludge Application on Soil Populations of Bradyrhizobium japonicum. AB - The application of sewage sludge to land may increase the concentration of heavy metals in soil. Of considerable concern is the effect of heavy metals on soil microorganisms, especially those involved in the biocycling of elements important to soil productivity. Bradyrhizobium japonicum is a soil bacterium involved in symbiotic nitrogen fixation with Glycine max, the common soybean. To examine the effect of metal-rich sludge application on B. japonicum, the MICs for Pb, Cu, Al, Fe, Ni, Zn, Cd, and Hg were determined in minimal media by using laboratory reference strains representing 11 common serogroups of B. japonicum. Marked differences were found among the B. japonicum strains for sensitivity to Cu, Cd, Zn, and Ni. Strain USDA 123 was most sensitive to these metals, whereas strain USDA 122 was most resistant. In field studies, a silt loam soil amended 11 years ago with 0, 56, or 112 Mg of digested sludge per ha was examined for total numbers of B. japonicum by using the most probable number method. Nodule isolates from soybean nodules grown on this soil were serologically typed, and their metal sensitivity was determined. The number of soybean rhizobia in the sludge-amended soils was found to increase with increasing rates of sludge. Soybean rhizobia strains from 11 serogroups were identified in the soils; however, no differences in serogroup distribution or proportion of resistant strains were found between the soils. Thus, the application of heavy metal-containing sewage sludge did not have a long-term detrimental effect on soil rhizobial numbers, nor did it result in a shift in nodule serogroup distribution. PMID- 16347282 TI - Selection of Streptococcus lactis Mutants Defective in Malolactic Fermentation. AB - An enrichment medium and a new sensitive medium were developed to detect malolactic variants in different strains of lactic bacteria. Factors such as the concentration of glucose and l-malate, pH level, and the type of indicator dye used are discussed with regard to the kinetics of malic acid conversion to lactic acid. Use of these media allowed a rapid and easier screening of mutagenized streptococcal cells unable to ferment l-malate. A collection of malolactic negative mutants of Streptococcus lactis induced by UV, nitrosoguanidine, or transposonal mutagenesis were characterized. The results showed that several mutants were apparently defective in the structural gene of malolactic enzyme, whereas others contained mutations which may either inactivate a putative permease or affect a regulatory sequence. PMID- 16347283 TI - Bacterial communities in acidic and circumneutral streams. AB - The relationship between pH and the abundance and activity of bacteria in streams was examined as part of a study of the effect of acidification on stream communities. Of the bacterial communities examined, the epilithic community appeared to be the most significantly affected by acidification. Microbial biomass, as quantified by measuring the ATP level, on rock surfaces was significantly correlated with pH. Also, bacterial production by the epilithic bacteria, indicated by incorporation of tritiated thymidine into DNA, was always higher at high-pH sites than at low-pH sites of the same stream order and elevation. Bacterioplankton concentrations varied between 0.53 x 10 and 9.42 x 10 cells . ml in the first- to fourth-order streams examined. The bacterioplankton concentration in one sample from a spring was 0.17 x 10 cells . ml. Bacterioplankton concentrations were not correlated with pH but were significantly correlated with seston concentrations. The correlation with seston is a result of increases in particle-associated bacteria at high seston concentrations. The proportion of bacterioplankton attached to particles varied from 0 to 70%. Bacterial numbers and production in the sediments were significantly correlated with the organic content of the sediment rather than with the pH of the overlying water. Thus, reduced abundance and activity of bacteria as a result of acidification could be detected only for the relatively active community on rock surfaces; this community was exposed to the low pH because of the unbuffered nature of its environment. PMID- 16347284 TI - Enzyme-Linked Immunosorbent Assay for Specific Identification and Enumeration of Azospirillum brasilense Cd. in Cereal Roots. AB - The enzyme-linked immunosorbent assay is suggested as a reliable, sensitive, and highly specific method for the identification and enumeration of Azospirillum brasilense Cd. As few as 10 CFU/ml can be practically identified by this method. At higher bacterial numbers, sensitivity increased linearly up to 5 x 10 CFU/ml, yielding useful standard curves. No cross-reaction was found either with different closely related Azospirillum strains or with other rhizosphere bacteria. The method allows for a specific identification of A. brasilense Cd. both in pure cultures and in mixtures with other bacterial species, even when the colony morphology is variable. The method was successfully applied to assess the degree of root colonization on various cereals by A. brasilense Cd. PMID- 16347285 TI - Flowthrough reactor flasks for study of microbial metabolism in sediments. AB - Flowthrough reactor flasks are described that allow continuous low-level nutrient input to mixed anoxic sediments without dilution of the sediment. The flasks were tested by simulating sulfate inputs into sediments collected from a freshwater eutrophic lake. After an initial 2-day adaptation within the reactor system, rates of methane production and sulfate consumption were constant for the duration of a 12-day incubation. A sulfate input rate of 0.15 mmol liter of sediment day resulted in an equivalent rate of sulfate removal, which was unaffected by inputs of acetate (1.0 mmol liter of sediment day). The rate of methane production in control reactors, 0.18 mmol liter of sediment day, was doubled by the addition of acetate, whereas sulfate consumption was only stimulated by additions of high concentrations of sulfate plus acetate (1.5 and 1.0 mmol liter of sediment day, respectively). The reactor system appears to be effective in maintaining the balance between sulfate reduction and methane production in freshwater sediments and is potentially useful for study of the response of sediment populations to varying inputs of naturally occurring substrates, selected inhibitors, or xenobiotic compounds. PMID- 16347286 TI - Bacteriophage Resistance Plasmid pTR2030 Inhibits Lytic Infection of r(1)t Temperate Bacteriophage but Not Induction of r(1)t Prophage in Streptococcus cremoris R1. AB - The effects of pTR2030 on the replication of four small isometric bacteriophages were examined in Streptococcus cremoris R1. Three lytic phages (652, 720, and 751), which were isolated independently over a 29-year period, were unable to form plaques on a pTR2030 transconjugant of S. cremoris R1. The fourth phage evaluated, phage r(1)t, was a temperate phage induced from S. cremoris R1 by treatment with mitomycin C. A prophage-cured derivative of S. cremoris R1, designated R1Cs, was isolated and served as a lytic indicator for phage r(1)t. Strain R1Cs and a derivative of this strain that was relysogenized with r(1)t, designated R1Cs(r(1)t), were used as conjugal recipients for transfer of the phage resistance plasmid pTR2030. pTR2030 transconjugants of strains R1Cs and R1Cs(r(1)t) were evaluated for sensitivity to r(1)t phage and induction of r(1)t prophage, respectively. The temperate phage r(1)t adsorbed eficiently but did not form plaques on the prophage-cured, pTR2030 transconjugant strain T-R1Cs. However, in the r(1)t lysogen [T-R1Cs(r(1)t)], pTR2030 did not inhibit prophage induction with mitomycin C, cell lysis, or production of infective r(1)t phage particles. The data demonstrated that pTR2030-induced resistance inhibited lytic infection by r(1)t phage from without but did not retard lytic development after prophage induction within the cell. It was suggested that pTR2030-encoded phage resistance to small isometric phages may, therefore, act at the cell surface or membrane to prevent phage DNA passage into the host cell or inhibit early events required for lytic replication of externally infecting phage. PMID- 16347287 TI - Rhizobium Population Genetics: Enzyme Polymorphism in Rhizobium leguminosarum from Plants and Soil in a Pea Crop. AB - A population of Rhizobium leguminosarum biovar viceae symbiotic on the roots of a commercial pea (Pisum sativum cv. Maro) crop was sampled by extracting a total of 249 isolates from root nodules on nine plants. Another 104 isolates were obtained by using soil from the same site to inoculate test plants, and a further 86 isolates were similarly obtained from soil 20 m distant within the crop. Each isolate was characterized for mobility variants of the enzymes glucose-6 phosphate dehydrogenase, superoxide dismutase, and beta-galactosidase by polyacrylamide gel electrophoresis. All three enzymes were polymorphic, and there was a strong disequilibrium among them. Of the 15 observed combinations of alleles (electrophoretic types [ETs]), 12 were indistinguishable from those previously described for isolates from a site 25 km distant. ET frequencies were significantly different among isolates from nodules on primary roots as opposed to lateral roots. The population on each individual plant was very diverse, but ET frequencies were similar from plant to plant. The ETs nodulating the primary roots were almost, although not perfectly, mixed, since the incidence of the same ETs in adjacent nodules was only about twice that expected by chance. The two samples derived from soil had the same ET frequencies but were significantly different from the field nodule sample, although the level of diversity was similar and there were no new ETs. PMID- 16347288 TI - Inhibition of methanogenesis from acetate in granular sludge by long-chain Fatty acids. AB - The effect of four saturated long-chain fatty acids (caprylic, capric, lauric, and myristic) and one unsaturated long-chain fatty acid (oleic) on the microbial formation of methane from acetate was investigated in batch anaerobic toxicity assays. The tests were carried out with granular sludge from an upflow anaerobic sludge bed reactor. In this sludge, Methanothrix spp. are the predominant acetoclastic methanogens. Lauric acid appeared to be the most versatile inhibitor: inhibition started at 1.6 mM, and at 4.3 mM the maximum specific acetoclastic methanogenic activity had been reduced to 50%. Caprylic acid appeared to be only slightly inhibitory. Oleic acid was almost as inhibitory as lauric acid. Although adsorption of the inhibitor on the cell wall might play an important role in the mechanism of inhibition, the inhibition was found to be correlated with concentration rather than with the amount per unit of biomass. In practical situations, as in anaerobic waste treatment processes, synergism can be expected to enhance the inhibition of methanogenesis. In the present research a background concentration of lauric acid below its MIC strongly enhanced the toxicity of capric acid and (to an even greater extent) myristic acid. PMID- 16347289 TI - Transposon Mutagenesis of Azospirillum brasilense and Azospirillum lipoferum: Physical Analysis of Tn5 and Tn5-Mob Insertion Mutants. AB - Tn5-induced insertion mutants were generated in Azospirillum brasilense Sp7 and A. lipoferum SpBr17 by mating with Escherichia coli strains carrying suicide plasmid vectors. The sources of Tn5 were the suicide plasmids pGS9 and pSUP2021. Kanamycin-resistant Azospirillum colonies appeared from crosses with E. coli at maximum frequencies of 10 per recipient cell. Transposon Tn5 also conferred streptomycin resistance on Azospirillum colonies as was observed earlier for Rhizobium sp. Eight Tn5-induced Km SmA. brasilense Sp7 mutants with reduced nitrogen-fixing capacity were isolated. The potential use of Tn5-Mob for labeling and mobilization of Azospirillum-indigenous plasmids was demonstrated by isolating Tn5-Mob insertions in the megaplasmids of A. brasilense Sp7. PMID- 16347290 TI - Toxic Trypsin Digest Fragment from the Bacillus thuringiensis Parasporal Protein. AB - Enzymatic digestion in vitro of the Bacillus thuringiensis protoxin presumably releases and activates the toxin in a manner analogous to that which occurs when a B. thuringiensis sporulated fermentation preparation passes through the midgut of a lepidopteran larva. Therefore, a sporulated culture of B. thuringiensis subsp. kurstaki (serotype 3a3b) HD-263 was treated with trypsin to release an activated toxin soluble in bicarbonate buffer. A 63-kilodalton protein, toxic to cabbage looper larvae (Trichoplusia ni) and to lepidopteran cells in culture, was purified to homogeneity from this trypsin digest. The larvicide, a glycoprotein containing 5% carbohydrate (wt/wt), was purified from the soluble B. thuringiensis trypsin digest by using ammonium sulfate precipitation, anion exchange chromatography, and hydrophobic-interaction chromatography. Its amino acid composition was high in nonpolar residues and unusually low in lysine and histidine. The isoelectric point was 6.5, and the amino acid on the N terminus was identified as isoleucine. The toxin was only slightly soluble in aqueous buffers unless the chaotropic agent potassium thiocyanate was added. Partial characterization of the toxin indicated that it corresponds well with reported sequences deduced from cloned genes. PMID- 16347291 TI - Symbiotic Expression of Cosmid-Borne Bradyrhizobium japonicum Hydrogenase Genes. AB - The expression of cosmid-borne Bradyrhizobium japonicum hydrogenase genes in alfalfa, clover, and soybean nodules harboring Rhizobium transconjugants was studied. Cosmid pHU52 conferred hydrogen uptake (Hup) activity in both free living bacteria and in nodules on the different plant hosts, although in nodules the instability of the cosmid resulted in low levels of Hup activity. In contrast, cosmid pHU1, which does not confer Hup activity on free-living bacteria, gave a Hup phenotype in nodules on alfalfa and soybean. Nodules formed by B. japonicum USDA 123Spc(pHU1) recycled about 90% of nitrogenase-mediated hydrogen evolution. Both subunits of hydrogenase (30- and 60-kilodalton polypeptides) were detected in enzyme-linked immunosorbent assays of bacteroid preparations from nodules harboring B. japonicum strains with pHU1 or pHU52. Neither pHU53 nor pLAFR1 conferred detectable Hup activity in either nodules or free-living bacteria. Based on the physical maps of pHU1 and pHU52, it is suggested that a 5.5-kilobase EcoRI fragment unique to pHU52 contains a gene or part of a gene required for Hup activity in free-living bacteria but not in nodules. This conclusion is supported by the observation that two Tn5 insertions in the chromosome of B. japonicum USDA 122DES obtained by marker exchange with Tn5-mutagenized pHU1 abolished Hup activity in free-living bacteria but not in nodules. PMID- 16347292 TI - Thermophilic anaerobic degradation of butyrate by a butyrate-utilizing bacterium in coculture and triculture with methanogenic bacteria. AB - We studied syntrophic butyrate degradation in thermophilic mixed cultures containing a butyrate-degrading bacterium isolated in coculture with Methanobacterium thermoautotrophicum or in triculture with M. thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic bacterium. Butyrate was beta-oxidized to acetate with protons as the electron acceptors. Acetate was used concurrently with its production in the triculture. We found a higher butyrate degradation rate in the triculture, in which both hydrogen and acetate were utilized, than in the coculture, in which acetate accumulated. Yeast extract, rumen fluid, and clarified digestor fluid stimulated butyrate degradation, while the effect of Trypticase was less pronounced. Penicillin G, d cycloserine, and vancomycin caused complete inhibition of butyrate utilization by the cultures. No growth or degradation of butyrate occurred when 2 bromoethanesulfonic acid or chloroform, specific inhibitors of methanogenic bacteria, was added to the cultures and common electron acceptors such as sulfate, nitrate, and fumarate were not used with butyrate as the electron donor. Addition of hydrogen or oxygen to the gas phase immediately stopped growth and butyrate degradation by the cultures. Butyrate was, however, metabolized at approximately the same rate when hydrogen was removed from the cultures and was metabolized at a reduced rate in the cultures previously exposed to hydrogen. PMID- 16347293 TI - Kinetics of butyrate, acetate, and hydrogen metabolism in a thermophilic, anaerobic, butyrate-degrading triculture. AB - Kinetics of butyrate, acetate, and hydrogen metabolism were determined with butyrate-limited, chemostat-grown tricultures of a thermophilic butyrate utilizing bacterium together with Methanobacterium thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic rod. Kinetic parameters were determined from progress curves fitted to the integrated form of the Michaelis-Menten equation. The apparent half-saturation constants, K(m), for butyrate, acetate, and dissolved hydrogen were 76 muM, 0.4 mM, and 8.5 muM, respectively. Butyrate and hydrogen were metabolized to a concentration of less than 1 muM, whereas acetate uptake usually ceased at a concentration of 25 to 75 muM, indicating a threshold level for acetate uptake. No significant differences in K(m) values for butyrate degradation were found between chemostat- and batch grown tricultures, although the maximum growth rate was somewhat higher in the batch cultures in which the medium was supplemented with yeast extract. Acetate utilization was found to be the rate-limiting reaction for complete degradation of butyrate to methane and carbon dioxide in continuous culture. Increasing the dilution rate resulted in a gradual accumulation of acetate. The results explain the low concentrations of butyrate and hydrogen normally found during anaerobic digestion and the observation that acetate is the first volatile fatty acid to accumulate upon a decrease in retention time or increase in organic loading of a digestor. PMID- 16347294 TI - Characterization of a Copper Resistance Plasmid Conserved in Copper-Resistant Strains of Pseudomonas syringae pv. tomato. AB - A 35-kilobase plasmid was conserved among 12 copper-resistant strains of Pseudomonas syringae pv. tomato. Restriction patterns of this plasmid from each strain were identical, and a cloned copper resistance gene from 1 strain hybridized to the same location on the 35-kilobase plasmid of all 12 strains. PMID- 16347295 TI - Ethanol Perturbs Glycosylation and Inhibits Hypersecretion in Trichoderma reesei. AB - The effects of ethanol and phenylethanol on the growth of and glycoprotein secretion by Trichoderma reesei were studied. Low levels (1.5%, vol/vol) of ethanol perturbed the glycosylation process, as shown by alterations in the isoelectric profile of the secreted proteins and a reduction in the rate of incorporation of mannose into oligosaccharides. In addition to these effects on posttranslational modification, ethanol drastically lowered the protein secretion level of a hypersecretory strain. PMID- 16347296 TI - Bacterial growth in mixed cultures on dissolved organic carbon from humic and clear waters. AB - Interactions between bacterial assemblages and dissolved organic carbon (DOC) from different sources were investigated. Mixed batch cultures were set up with water from a humic and a clear-water lake by a 1:20 dilution of the bacterial assemblage (1.0 mum of prefiltered lake water) with natural medium (sterile filtered lake water) in all four possible combinations of the two waters and their bacterial assemblages. Bacterial numbers and biomass, DOC, thymidine incorporation, ATP, and uptake of glucose and phenol were followed in these cultures. Growth curves and exponential growth rates were similar in all cultures, regardless of inoculum or medium. However, bacterial biomass produced was double in cultures based on water from the humic lake. The fraction of DOC consumed by heterotrophic bacteria during growth was in the same range, 15 to 22% of the total DOC pool, in all cultures. Bacterial growth efficiency, calculated from bacterial biomass produced and DOC consumed, was in the order of 20%. Glucose uptake reached a peak during exponential growth in all cultures. Phenol uptake was insignificant in the cultures based on the clear-water medium, but occurred in humic medium cultures after exponential growth. The similarity in the carbon budgets of all cultures indicated that the source of the bacterial assemblage did not have a significant effect on the overall carbon flux. However, fluxes of specific organic compounds differed, as reflected by glucose and phenol uptake, depending on the nature of the DOC and the bacterial assemblage. PMID- 16347297 TI - Growth of Azotobacter vinelandii on Soil Nutrients. AB - Azotobacter vinelandii cells grew well in a medium made from soil and distilled water which contained little or no carbohydrate. They utilized p-hydroxybenzoic acid and other phenolic acids, soil nitrogen, and water-soluble mineral substances. Seventeen soils which supported excellent growth of A. vinelandii contained 11 to 18 different phenolic acids each, including p-hydroxybenzoic, m hydroxybenzoic, vanillic, p-coumeric, syringic, cis- and trans-ferrulic, and other unidentified aromatic acids. Three white, chalky "caliche" soils which were taken from areas where no plants grew failed to support the growth of A. vinelandii, and these contained no, two, and three phenolic acids, respectively. A. vinelandii did not fix nitrogen when growing in dialysates of soils which contained numerous phenolic acids. Growth was ample and rapid in most of the soils tested, but cell morphology was different from that usually seen in chemically defined, nitrogen-free media which contain glucose. PMID- 16347298 TI - Applicability of yeast extracellular proteinases in brewing: physiological and biochemical aspects. AB - A general screening survey for expression of extracellular acid proteinase production was performed on over 100 cultures belonging to the genus Saccharomyces. Although two strains of Saccharomyces cerevisiae showed positive extracellular proteinase phenotypes in plate tests, it was not possible to demonstrate proteolytic activities in cell-free culture supernatants in assays performed at beer pH values. Of several yeasts from other genera examined, Saccharomycopsis fibuligera and Torulopsis magnoliae produced extracellular proteinases with desirable properties. Proteolytic activities were detected in assays performed at beer pH values and at lower temperature. Brewer's wort served as a highly inducing medium for extracellular proteinase production, with T. magnoliae yielding enzyme of highest specific activity. In fact, commencement of enzyme production was detected shortly after the onset of exponential growth in brewer's wort. Inclusion of crude enzyme preparations in brewer's wort inoculated simultaneously with brewer's yeast reduced final ethanol yields slightly and was found to be effective in reducing chill haze formation in bottled beer. PMID- 16347299 TI - Specificity of Bacillus thuringiensis Delta-Endotoxin. AB - The insecticidal activity of the delta-endotoxins of 14 Bacillus thuringiensis strains belonging to 12 subspecies was determined against Pieris brassicae, Heliothis virescens, and Spodoptera littoralis. Larvae of P. brassicae were highly susceptible to purified crystals of strains of B. thuringiensis subsp. thuringiensis and B. thuringiensis subsp. morrisoni, whereas H. virescens responded best to B. thuringiensis subsp. kenyae and B. thuringiensis subsp. kurstaki. The crystals of the B. thuringiensis subsp. entomocidus strain were the most potent against S. littoralis. It was shown that the solubility of the crystals within the gut of the three insect species is a first important step in the mode of action. Predissolution of the crystals especially enhanced the insecticidal activity against H. virescens. When in vitro-activated toxins were applied, the relative potency range varied greatly from one insect species to another. It can be concluded that at least three factors influence the potency of B. thuringiensis delta-endotoxins: the strain-related origin of the toxin, the degree of solubility of the crystals in the gut juice, and the intrinsic susceptibility of the insect to the toxin. PMID- 16347300 TI - Lysis of Bacillus subtilis Cells by Glycerol and Sucrose Esters of Fatty Acids. AB - The lytic action of glycerol and sucrose esters of fatty acids with different carbon chain lengths on the exponentially growing cells of Bacillus subtilis 168 was investigated. Of each series of esters, glycerol dodecanoate and sucrose hexadecanoate were the most active. Lysis at 1 h after the addition of 0.1 mM glycerol dodecanoate or 20 mug of sucrose hexadecanoate per ml was 81 or 79%, respectively, as evaluated by the reduction in optical density. During this treatment a great loss of viability occurred that preceded lysis. The results that were obtained suggest that autolysis is induced by these esters. The esters caused morphological changes in the cells, but a seeming adaptation of the cells to esters was seen. PMID- 16347301 TI - One-Step Microbial Conversion of a Racemic Mixture of Pantoyl Lactone to Optically Active d-(-)-Pantoyl Lactone. AB - Washed cells of Rhodococcus erythropolis IFO 12540 were found to convert only the l-(+)-isomer of pantoyl lactone to the d-(-)-isomer in a racemic mixture of pantoyl lactone. Under suitable reaction conditions, the amount of d-(-)-pantoyl lactone synthesized was 18.2 mg/ml (94.4% enantiomer excess; molar yield, 90.5%). This conversion was suggested to proceed through the following successive reactions: first, the enzymatic oxidation of l-(+)-pantoyl lactone to ketopantoyl lactone; second, the rapid and spontaneous hydrolysis of the ketopantoyl lactone to ketopantoic acid; and then, the enzymatic reduction of the ketopantoic acid to d-(-)-pantoic acid. After the reaction d-(-)-pantoic acid could be lactonized by means of acid treatment. During the conversion, the d-(-)-isomer, which was initially present in the reaction mixture, did not undergo any modification. PMID- 16347302 TI - Properties of the glucose transport system in some deep-sea bacteria. AB - Many deep-sea bacteria are specifically adapted to flourish under the high hydrostatic pressures which exist in their natural environment. For better understanding of the physiology and biochemistry of these microorganisms, properties of the glucose transport systems in two barophilic isolates (PE-36, CNPT-3) and one psychrophilic marine bacterium (Vibrio marinus MP1) were studied. These bacteria use a phosphoenol-pyruvate:sugar phosphotransferase system (PTS) for glucose transport, similar to that found in many members of the Vibrionaceae and Enterobacteriaceae. The system was highly specific for glucose and its nonmetabolizable analog, methyl alpha-glucoside (a-MG), and exhibited little affinity for other sugars tested. The temperature optimum for glucose phosphorylation in vitro was approximately 20 degrees C. Membrane-bound PTS components of deep-sea bacteria were capable of enzymatically cross-reacting with the soluble PTS enzymes of Salmonella typhimurium, indicating functional similarities between the PTS systems of these organisms. In CNPT-3 and V. marinus, increased pressure had an inhibitory effect on a-MG uptake, to the greatest extent in V. marinus. Relative to atmospheric pressure, increased pressure stimulated sugar uptake in the barophilic isolate PE-36 considerably. Increased hydrostatic pressure inhibited in vitro phosphoenolpyruvate-dependent a MG phosphorylation catalyzed by crude extracts of V. marinus and PE-36 but enhanced this activity in crude extracts of the barophile CNPT-3. Both of the pressure-adapted barophilic bacteria were capable of a-MG uptake at higher pressures than was the nonbarophilic psychrophile, V. marinus. PMID- 16347303 TI - Effect of oxygen on lactose metabolism in lactic streptococci. AB - Three strains of Streptococcus lactis, two of Streptococcus cremoris, and one of Streptococcus thermophilus metabolized oxygen in the presence of added carbohydrate primarily via a closely coupled NADH oxidase/NADH peroxidase system. No buildup of the toxic intermediate H(2)O(2) was detected with the three S. lactis strains. All six strains contained significant superoxide dismutase activity and are clearly aerotolerant. Lactose- or glucose-driven oxygen consumption was biphasic, with a rapid initial rate followed by a slower secondary rate which correlated with factors affecting the in vivo activation of lactate dehydrogenase. The rate of oxygen consumption was rapid under conditions that led to a reduction in lactate dehydrogenase activity (low intracellular fructose 1,6-bisphosphate concentration). These conditions could be achieved with nongrowing cells by adding lactose at a constant but limiting rate. When the rate of lactose fermentation was limited to 5% of its maximum, nongrowing cells of S. lactis strains ML3 and ML8 carried out an essentially homoacetic fermentation under aerobic conditions. These same cells carried out the expected homolactic fermentation when presented with excess lactose under anaerobic conditions. Homoacetic fermentation leads to the generation of more energy, by substrate level phosphorylation via acetate kinase, than the homolactic fermentation. However, it was not observed in growing cells and was restricted to slow fermentation rates with nongrowing cells. PMID- 16347304 TI - Conjugal Transfer of Plasmid-Encoded Determinants for Bacteriocin Production and Immunity in Lactobacillus acidophilus 88. AB - Lactobacillus acidophilus 88 produced a bacteriocin, designated lactacin F, that demonstrated inhibitory activity toward L. acidophilus 6032, L. lactis 970, L. helveticus 87, L. bulgaricus 1489, L. leichmanii 4797, L. fermentum 1750, and Streptococcus faecalis 19433. Production of lactacin F was pH dependent and could be maximized in MRS broth cultures maintained at pH 7.0. Lactacin F was heat stable and sensitive to ficin, proteinase K, trypsin, and Bacillus subtilis protease. L. acidophilus 88 harbored plasmids of 4 and 27 megadaltons. Variants of L. acidophilus 88 which were deficient in lactacin F production (Laf) and lactacin F immunity (Laf) retained the two resident plasmids. A Laf Laf derivative, L. acidophilus 89, was used as a recipient in agar surface mating experiments with L. acidophilus 88 (Laf Laf). Two types of Laf Laf transconjugants were recovered. One type (T-E) had acquired two plasmids of 68 (pPM68) and 52 (pPM52) megadaltons that were not detected in either the conjugal donor or the other type of Laf Laf transconjugants (T-89). Laf and Laf were unstable in the plasmid-bearing transconjugant. Plasmid analysis of Laf Laf variants revealed that pPM52 and pPM68 were cured with loss of Laf and Laf. Bacteriocin production and immunity phenotypes were genetically stable in Laf Laf transconjugants not harboring pPM52 and pPM68, suggesting chromosomal integration of the transferred determinants. The data demonstrated intragenic conjugation in L. acidophilus and provided direct evidence for involvement of transient plasmid determinants in Laf and Laf. PMID- 16347305 TI - Analysis of yeast flora associated with grape sour rot and of the chemical disease markers. AB - The frequency and the density of the species associated with grape sour rot in different cultivars were determined. The most frequent species in the rotten grapes, Candida krusei, Kloeckera apiculata, and Metschnikowia pulcherrima, and a less frequent species, Issatchenkia occidentalis, when inoculated with Saccharomycopsis crataegensis were able to induce in vitro the symptoms of the disease. The gas chromatographic determination of the volatile compounds in the headspace was used to evaluate the metabolic role of the different species associated with the disease. These analyses made it possible to presume that, whereas some species, such as Candida krusei and Hanseniaspora uvarum, can be considered responsible for these modifications and in particular for the ethyl acetate production, others, such as Saccharomycopsis crataegensis, can promote the development of the former species. PMID- 16347306 TI - Purification and Some Properties of a Membrane-Bound Aminopeptidase A from Streptococcus cremoris. AB - A membrane-bound l-alpha-glutamyl (aspartyl)-peptide hydrolase (aminopeptidase A) (EC 3.4.11.7) from Streptococcus cremoris HP has been purified to homogeneity. The free gamma-carboxyl group rather than the amino group of the N-terminal l alpha-glutamyl (aspartyl) residue appeared to be essential for catalysis. No endopeptidase activity could be established with this enzyme. The native enzyme is a polymeric, most probably trimeric, metalloenzyme (relative molecular weight, approximately 130,000) which shows on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels apparent high relative molecular weight values due to (lipid?) material dissociable with butanol. The subunit (relative molecular weight, approximately 43,000) is catalytically inactive. The enzyme is inactivated completely by dithiothreitol, chelating agents, and the bivalent metal ions Cu and Hg. Of the sulfhydryl-blocking reagents tested, only p hydroxymercuribenzoate appeared to inhibit the enzyme. Activity lost by treatment with a chelating agent could be restored by Co and Zn. The importance of the occurrence of an aminopeptidase A in S. cremoris with respect to growth in milk is discussed. PMID- 16347307 TI - Influence of Soil and Nonsoil Environments on Nodulation by Rhizobium trifolii. AB - Indigenous serotypes 1-01 and 2-02 of Rhizobium trifolii occupied similar percentages (18 to 23%) of root nodules on soil-grown subclover (Trifolium subterraneum L.) and were virtually absent (4.5%) from nodules of soil-grown white clover (Trifolium repens L.). In contrast (with the exception of one dilution [10]), serotype 1-01 occupied a substantial portion of nodules (16 to 40%) on white clover seedlings grown on mineral salts agar and exposed to samples of the same soil in the form of a 10-fold dilution series (10 to 10). Under the latter conditions, occupancy of subclover nodules by 1-01 and of nodules of both plant species by 2-02 was consistent with the results obtained with soil-grown plants. PMID- 16347308 TI - Measurement of surface charge of baculovirus polyhedra. AB - The isoelectric points of three baculoviruses, Trichoplusia ni nuclear polyhedrosis virus (NPV), T. ni granulosis virus, and Spodoptera littoralis NPV were identified by cell electrophoresis. At neutral pH polyhedra were negatively charged. T. ni NPV polyhedra were reacted with a number of reagents which could potentially attach to or degrade their surface structure. This gave information on the components that contribute to the charge profile of T. ni NPV. This is discussed in relation to the use of polyhedra as biological control agents against insect pests. PMID- 16347309 TI - Influence of the Bradyrhizobium japonicum Hydrogenase on the Growth of Glycine and Vigna Species. AB - The effect of the Bradyrhizobium japonicum hydrogenase on nitrogen fixation was evaluated by comparing the growth of Vigna and Glycine species inoculated with a Hup mutant and its Hup revertant. In all experiments, the growth of plants inoculated with the strain without hydrogenase was at least equal to the growth of the strain with hydrogenase. For Glycine usuriensis and Glycine max cv. Hodgson in liquid culture, the growth was higher with the Hup strain. It is possible that reduced rates of nitrogen fixation in the presence of hydrogenase are due to O(2) depletion caused by the hydrogen oxidizing, since the oxygen pressure in the air appears to be a limiting factor of symbiotic nitrogen fixation in the soybean. PMID- 16347310 TI - Microplate assay for colletotrichum spore production. AB - A simple microplate method was devised to assay spore production by Colletotrichum gloeosporioides by growing the fungus on 1 ml of solid media in the wells of tissue culture plates. Growth and sporulation on microplates were compared at days 4 and 8 with growth and sporulation in 100-ml liquid batch cultures that involved 11 common media. Spore production per unit volume of medium was the same for solid and liquid forms of the media. Qualitative assessment of mycelial growth measured on microplates agreed with that of growth measured in liquid cultures. The microplate assay indicated that V8 juice was the best medium and that an organic content of about 6 mg/ml was optimal for high sporulation and low mycelium production. The assay provides a convenient, rapid, and inexpensive means of screening media for the production of fungal conidia in large numbers, to be used, for example, in biological control programs. PMID- 16347311 TI - Production of 1,3-Propanediol from Glycerol by Clostridium acetobutylicum and Other Clostridium Species. AB - Glycerol was fermented with the production of 1,3-propanediol as the major fermentation product by four strains of Clostridium acetobutylicum, six of C. butylicum, two of C. beijerinckii, one of C. kainantoi, and three of C. butyricum. 1,3-Propanediol was identified by its retention times in gas chromatography and high-pressure liquid chromatography and by its mass spectrum. During growth of C. butylicum B593 in a chemostat culture at pH 6.5, 61% of the glycerol fermented was converted to 1,3-propanediol. When the pH was decreased to 4.9, growth and 1,3-propanediol production were substantially reduced. PMID- 16347312 TI - Purification and Characterization of Two Endoxylanases from Clostridium acetobutylicum ATCC 824. AB - Two endoxylanases produced by C. acetobutylicum ATCC 824 were purified to homogeneity by column chromatography. Xylanase A, which has a molecular weight of 65,000, hydrolyzed larchwood xylan randomly, yielding xylohexaose, xylopentaose, xylotetraose, xylotriose, and xylobiose as end products. Xylanase B, which has a molecular weight of 29,000, also hydrolyzed xylan randomly, giving xylotriose and xylobiose as end products. Xylanase A hydrolyzed carboxymethyl cellulose with a higher specific activity than xylan. It also exhibited high activity on acid swollen cellulose. Xylanase B showed practically no activity against either cellulose or carboxymethyl cellulose but was able to hydrolyze lichenan with a specific activity similar to that for xylan. Both xylanases had no aryl-beta xylosidase activity. The smallest oligosaccharides degraded by xylanases A and B were xylohexaose and xylotetraose, respectively. The two xylanases demonstrated similar K(m) and V(max) values but had different pH optima and isoelectric points. Ouchterlony immunodiffusion tests showed that xylanases A and B lacked antigenic similarity. PMID- 16347313 TI - Isolation and Some Properties of a beta-d-Xylosidase from Clostridium acetobutylicum ATCC 824. AB - A beta-d-xylosidase from C. acetobutylicum ATCC 824 was purified by column chromatography on CM-Sepharose, hydroxylapatite, Phenyl Sepharose, and Sephadex G 200. The enzyme had an apparent molecular weight of 224,000 as estimated by gel filtration. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the enzyme consisted of two subunits of 85,000 and one subunit of 63,000 daltons. It exhibited optimal activity at pH 6.0 to 6.5 and 45 degrees C. the enzyme had an isoelectric point of 5.85. It hydrolyzed p-nitrophenylxyloside readily with a K(m) of 3.7 mM. The enzyme hydrolyzed xylo-oligosaccharides with chain lengths of 2 to 6 units by cleaving a single xylose from the chain end. It showed little or no activity against xylan, carboxymethyl cellulose, and other p nitrophenylglycosides. PMID- 16347314 TI - Population Heterogeneity of Agrobacterium tumefaciens in Galls of Populus L. from a Single Nursery. AB - This study focused on the natural crown gall infections occurring in a Leuce poplar nursery. Soil effects on crown gall frequency were detected, indicating that contamination was due to a resident Agrobacterium tumefaciens population, which was present before seedling plantation. The crown gall frequency on poplar progenies varied from 3 to 67%, indicating the feasibility of improvement in crown gall resistance. Of 129 tumor isolates, 128 were pathogenic. These isolates were of biotype 1 or 2. Biochemical, serological, and antibiotic resistance typing results concurred, indicating the presence of four biotype 1 and two biotype 2 resident subpopulations. No significant change was noticed in the relative proportions of subpopulations from one year to another. Pathogenic subpopulations both in vitro and in planta were susceptible to Kerr K84 (P. B. New and A. Kerr, J. Appl. Bacteriol. 90:172-179, 1972). In addition, no serological cross-reactions were found to occur between K84 and the pathogenic subpopulations. PMID- 16347315 TI - A Combined Immunofluorescence-DNA-Fluorescence Staining Technique for Enumeration of Thiobacillus ferrooxidans in a Population of Acidophilic Bacteria. AB - An antiserum raised against whole cells of Thiobacillus ferrooxidans was allowed to react with a variety of acidophilic and nonacidophilic bacteria in an enzyme linked immunosorbent assay and an indirect immunofluorescence assay. Both experiments demonstrated that the antiserum was specific at the species level. This preparation was used to evaluate the role of T. ferrooxidans in the microbial desulfurization process. Leaching experiments were performed, and the numbers of T. ferrooxidans cells and other bacteria were estimated by using a combined immunofluorescence-DNA-fluorescence staining technique that was adapted for this purpose. Nonsterile coal samples inoculated with T. ferrooxidans yielded high concentrations of soluble iron after 16 days. After this period, however, T. ferrooxidans cells could no longer be detected by the immunofluorescence assay, whereas the DNA-fluorescence staining procedure demonstrated a large number of microorganisms on the coal particles. These results indicate that T. ferrooxidans is removed by competition with different acidophilic microorganisms that were originally present on the coal. PMID- 16347316 TI - Effect of Increased beta-Glucosidase Activity on Virulence of Erwinia amylovora. AB - Plant tissues often contain beta-glucosides that can be enzymatically hydrolyzed to produce toxic aglycones. It has been suggested that the low beta-glucosidase activity found in Erwinia amylovora contributes to bacterial virulence by allowing the bacteria to infect plants that contain beta-glucosides without inducing the formation of toxic aglycones. To test this suggestion, we created strains of E. amylovora which had high beta-glucosidase activities and studied the ability of these strains to cause fire blight disease in pears (Pyrus communis). We isolated spontaneous mutants that were able to utilize beta glucosides as the sole carbon source and showed that one class had about 10 times as much beta-glucosidase activity as the wild-type strain. In addition, we constructed several plasmids that carry the Escherichia coli bgl operon under the control of a transposon Tn5 promoter that is expressed in E. amylovora. These plasmids were introduced in E. amylovora by transformation. Pathogenesis studies in immature Bartlett pear fruits, etiolated sprouts, and young shoots showed that a 100-fold increase in beta-glucosidase activity does not interfere with normal development of fire blight disease in these model systems. PMID- 16347317 TI - Butanol-Ethanol Dehydrogenase and Butanol-Ethanol-Isopropanol Dehydrogenase: Different Alcohol Dehydrogenases in Two Strains of Clostridium beijerinckii (Clostridium butylicum). AB - Alcohol-producing strains of Clostridium beijerinckii (Clostridium butylicum) produce, besides acetone, either n-butanol and ethanol or n-butanol, ethanol, and isopropanol as their characteristic products. Alcohol dehydrogenase has been isolated from a strain (NRRL B593) of C. beijerinckii producing isopropanol and from a strain (NRRL B592) not producing isopropanol. Butanol-ethanol dehydrogenase activities were present in both strains, but isopropanol dehydrogenase activity was present only in the isopropanol-producing strain. The butanol-ethanol dehydrogenase of strain NRRL B592 had M(r) 66,000 and a K(m) of 6 muM for butyraldehyde. In contrast, the butanol-ethanol-isopropanol dehydrogenase of strain NRRL B593 had a M(r) 100,000 and K(m)s of 9.5 and 1.0 mM for butyraldehyde and acetone, respectively. In a purification by four different types of separatory methods (DEAE-cellulose, hydroxyapatite, Sephacryl S-300, and Matrex Gel Red A), butanol-ethanol-isopropanol dehydrogenase activities of strain NRRL B593 were purified up to 200-fold (10 to 30% yield), and these activities were not separated. Gel electrophoresis followed by activity stain also revealed distinct mobilities for the butanol-ethanol dehydrogenase of strain NRRL B592 and the butanol-ethanol-isopropanol dehydrogenase of strain NRRL B593. In cell extracts from both strains, a higher alcohol dehydrogenase activity was measured with NADP(H) than with NAD(H). The 150- to 200-fold-purified alcohol dehydrogenase from strain NRRL B593 did not show any NAD(H)-linked activities. The K(m) for NADPH was 31 muM (with butyraldehyde as cosubstrate) and 18 muM (with acetone as cosubstrate) for the alcohol dehydrogenase of strain NRRL B593. This study showed that the alcohol dehydrogenases from two strains of C. beijerinckii differed significantly. PMID- 16347318 TI - Isolation of different agrobacterium biovars from a natural oak savanna and tallgrass prairie. AB - Populations of agrobacteria in excess of 10 CFU/g were recovered from 12 soil and root samples obtained from the Allison Savanna, Minn., a natural oak savanna and tallgrass prairie which has never been disturbed agriculturally. Of 126 strains picked randomly from selective media, 54 were identified as Agrobacterium spp. Biovar 2 strains predominated (35 of 54), but these strains were distributed into three phenotypically distinct subgroups. Of the remaining Agrobacterium strains, four were biovar 1-2, one was biovar 1, and none were biovar 3. The last 14 Agrobacterium strains formed a homogeneous group which differed biochemically from the hitherto reported biovars. Opine utilization (coded for by genes on the tumor-inducing plasmid in pathogenic Agrobacterium spp.) by these agrobacteria was limited to two biovar 2 strains. In contrast, 10 nonfluorescent gram-negative strains utilized either nopaline or octopine as the sole carbon and nitrogen source. There may be a need to reexamine the source and role of opines in the terrestrial environment because (i) all of these opine utilizers were isolated from an environment free of crown gall, the only known terrestrial source of opines, and (ii) 83% of the opine utilizers were not Agrobacterium spp. PMID- 16347319 TI - Nitrifying bacteria in wastewater reservoirs. AB - Deep wastewater reservoirs are used throughout Israel to store domestic wastewater effluents for summer irrigation. These effluents contain high concentrations of ammonia (/= 10 muM. Pyoverdin(pss) was isolated, chemically characterized, and found to resemble previously characterized pyoverdins in spectral traits (absorbance maxima of 365 and 410 nm for pyoverdin(pss) and its ferric chelate, respectively), size (1,175 molecular weight), and amino acid composition. Nevertheless, pyoverdin(pss) was structurally unique since amino acid analysis of reductive hydrolysates yielded beta-hydroxyaspartic acid, serine, threonine, and lysine in a 2:2:2:1 ratio. Pyoverdin(pss) exhibited a relatively high affinity constant for Fe(III), with values of 10 at pH 7.0 and 10 at pH 10.0. Iron uptake assays with [Fe]pyoverdin(pss) demonstrated rapid active uptake of Fe(III) by P. syringae pv. syringae B301D, while no uptake was observed for a mutant strain unable to acquire Fe(III) from ferric pyoverdin(pss). The chemical and biological properties of pyoverdin(pss) are discussed in relation to virulence and iron uptake during plant pathogenesis. PMID- 16347353 TI - Isolation and characterization of thermophilic bacterial strains with inulinase activity. AB - Nine bacterial strains growing on inulin as the sole carbon and energy source were isolated from soil samples by enrichment culture on a mineral medium. Four of the strains were thermophilic and belong to the genus Bacillus. The thermophilic strains synthesized a beta-fructosidase that was active on both inulin and sucrose. The presence of inulin in the culture medium is necessary for enzyme synthesis. Most of the activity on inulin was recovered in the culture medium, and the enzyme was synthesized during cell growth. PMID- 16347354 TI - H(2)-CO(2) Recirculation and pH Control for Growth of Methanogens in Mass Culture. AB - We modified a fermentor (10-liter liquid volume) for the growth of anaerobic, H(2)-CO(2)-catabolizing bacteria. Gas in the fermentor (ca. 10% CO(2), 50% H(2), 40% CH(4)) was recirculated by a diaphragm pump. During growth, the gas composition was maintained by the addition of a mixture of 80% H(2) and 20% CO(2), and this addition was controlled by a pH auxostat. During gas addition, gas was discharged from the recirculating gas stream and was collected by the displacement of an acidified salt solution. PMID- 16347355 TI - Use of monodispersed, fluorescently labeled bacteria to estimate in situ protozoan bacterivory. AB - We have developed a procedure for preparing monodispersed, fluorescently labeled bacteria (FLB), which may be used to measure virtually instantaneous rates of protozoan bacterivory in natural waters. FLB can be prepared both from natural bacterioplankton assemblages and from clonal isolates and can be stored in frozen suspension or freeze-dried without apparent loss of fluorescence intensity. They are not toxic to protozoa and can be metabolized to support bacterivorous protozoan growth rates equal to those on the same strain of unstained, viable bacteria. In experiments comparing uptake of FLB with uptake of fluorescent latex microspheres by protozoan assemblages in a salt marsh tidal creek, we found that both pelagic oligotrichous ciliates and phagotrophic flagellates ingested FLB with a frequency 4- to 10-fold greater than they ingested the microspheres. Consequently, it appears that the use of latex microspheres leads to underestimation of protozoan bacterivory and that the FLB technique is superior for estimating instantaneous rates of in situ protozoan grazing on bacterioplankton. PMID- 16347356 TI - Uptake of Glyphosate by an Arthrobacter sp. AB - The uptake of glyphosate (N-[phosphonomethyl]glycine) by an Arthrobacter sp. which can utilize this herbicide as its sole source of phosphorus was investigated. Orthophosphate suppressed the expression of the uptake system for glyphosate and also competed with glyphosate for uptake. The K(m) for glyphosate uptake was 125 muM, and the K(i) for orthophosphate was 24 muM. Organophosphonates as well as organophosphates inhibited glyphosate uptake, but only organophosphates and orthophosphate suppressed the uptake system. Glyphosate uptake was energy dependent, had a pH optimum of 6 to 7, and was differentially affected by divalent cations. PMID- 16347357 TI - Altered Electron Flow in Continuous Cultures of Clostridium acetobutylicum Induced by Viologen Dyes. AB - The physiological response of Clostridium acetobutylicum to methyl and benzyl viologen was investigated. Viologen dyes at low concentrations (at levels of parts per million [micrograms per milliliter]) caused significant metabolic shifts. Altered electron flow appeared to direct carbon flow from acid to alcohol production accompanied by decreased hydrogen evolution. Reducing equivalents normally released as free hydrogen were directed toward formation of NADH which, in turn, resulted in increased alcohol production. In addition, it was shown that solvent production can take place at pH 6.3. Contrary to previous reports, butanol production appears to be independent of high levels of acetate-butyrate and glucose. PMID- 16347358 TI - Citrate Metabolism by Pediococcus halophilus. AB - Several strains of non-citrate-metabolizing Pediococcus halophilus have previously been isolated from soy sauce mash or moromi. The factors controlling the metabolism of citrate in soy pediococci were studied. All the soy pediococcal strains tested which failed to decompose citrate did not possess citrate lyase [citrate (pro-3S)-lyase; EC 4.1.3.6] activity. In P. halophilus, citrate lyase was an inducible enzyme, and the optimum pH for activity was 7.0. The metabolism of citrate in P. halophilus was different from that observed in lactic streptococci. The main products from citrate were acetate and formate, and this bacterium produced no acetoin or diacetyl. Formate production from citrate was greatly reduced in the presence of glucose. P. halophilus 7117 (Cit) was proved to contain citrate lyase, pyruvate formate-lyase (EC 2.3.1.54) phosphotransacetylase (phosphate acetyltransferase; EC 2.3.1.8), and acetate kinase (EC 2.7.2.1), i.e., all the enzymes necessary to convert citrate to acetate and formate. PMID- 16347359 TI - Selective Process for Efficient Isolation of Soil Bacillus spp. AB - We were able to isolate Bacillus thuringiensis from environmental samples with a background of 10 bacteria per g of soil. Our selection process differed significantly from classical selection methods which permit only the desired organism to grow. In our process, germination of B. thuringiensis spores was selectively inhibited by sodium acetate, while most of the undesired sporeformers germinated. Next, all of the nonsporulated microbes were eliminated by heat treatment at 80 degrees C for 3 min. The surviving spores were then plated on a rich agar medium and allowed to grow until they sporulated. Of random colonies picked from agar, 20 to 96% were crystal-forming Bacillus species. B. thuringiensis and B. sphaericus were routinely selected by this method. PMID- 16347360 TI - Annual bacterioplankton biomasses and productivities in a temperate west coast canadian fjord. AB - Bacterioplankton numbers, biomasses, and productivities, as well as chlorophyll a concentrations and phytoplankton productivities, were assayed from 1 March 1984 to 12 August 1985 through a 250-m-deep seawater column in Howe Sound, a temperate fjord-sound on the southern coast of British Columbia, Canada. Primary production during this 18-month period was 845 g of C m. Bacterial production was assayed over this same period as 193 g of C m (thymidine incorporation) and 77 g of C m (frequency of dividing cells). Bacterial productivities per cubic meter were usually greater in the euphotic zone than in deeper aphotic water, but when integrated through the water column, approximately half of the bacterial production occurred in the deeper aphotic portion. Bacterial production occurred throughout the year, although at reduced rates in late fall and early winter; primary production almost ceased during late fall and early winter. Because of this heterotrophic bacterioplankton production was a very large portion of the microbial (bacterial plus phyto-plankton) production at this time. In mid-summer bacterial production was a small proportion of the microbial production. Because of this asynchrony in peaks and troughs of bacterial and phytoplankton production through the year, data comparison is best done over an annual cycle. On this basis the bacterial production in the Howe Sound water column was between 23 and 9% of the phytoplankton production when a bacterial C to biovolume ratio of 0.107 pg of C mum was assumed; the corresponding values were 64 and 29% when a ratio of 0.300 pg of bacterial C mum was assumed. PMID- 16347361 TI - Denitrification in aquifer soil and nearshore marine sediments influenced by groundwater nitrate. AB - We estimated rates of denitrification at various depths in sediments known to be affected by submarine discharge of groundwater, and also in the parent aquifer. Surface denitrification was only measured in the autumn; at 40-cm depth, where groundwater-imported nitrate has been measured, denitrification occurred consistently throughout the year, at rates from 0.14 to 2.8 ng-atom of N g day. Denitrification consistently occurred below the zone of sulfate reduction and was sometimes comparable to it in magnitude. Denitrification occurred deep (14 to 40 cm) in the sediments along 30 km of shoreline, with highest rates occurring where groundwater input was greatest. Denitrification rates decreased with distance offshore, as does groundwater influx. Added glucose greatly stimulated denitrification at depth, but added nitrate did not. High rates of denitrification were measured in the aquifer (17 ng-atom of N g day), and added nitrate did stimulate denitrification there. The denitrification measured was enough to remove 46% of the nitrate decrease observed between 40- and 14-cm depth in the sediment. PMID- 16347362 TI - Relationships between Biovolume and Biomass of Naturally Derived Marine Bacterioplankton. AB - Microscopic estimation of bacterial biomass requires determination of both biovolume and biovolume-to-biomass conversion. Both steps have uncertainty when applied to the very small bacteria typically found in natural seawater. In the present study, natural bacterioplankton assemblages were freshly collected, passed through 0.6-mum-pore-size Nuclepore filters to remove larger particulate materials, and diluted for growth in 0.22-mum-pore-size Millipore filter sterilized unenriched seawater. This provided cells comparable in size and morphology to those in natural seawater, but the cultures were free of the interfering particulate detritus naturally present. Cells were collected on glass fiber GF/F filters, and biovolumes were corrected for cells passing these filters; C and N were measured with a CHN analyzer. Our criteria for size measurement by epifluorescence photomicrography were confirmed with fluorescent microspheres of known diameters. Surprisingly, in six cultures with average per cell biovolumes ranging from 0.036 to 0.073 mum, the average per-cell carbon biomass was relatively constant at 20 +/- 0.08 fg of C (mean +/- standard error of the mean). The biovolume-to-biomass conversion factor averaged 0.38 +/- 0.05 g of C cm, which is about three times higher than the value previously estimated from Escherichia coli, and decreased with increasing cell volume. The C:N ratio was 3.7 +/- 0.2. We conclude that natural marine bacterial biomass and production may be higher than was previously thought and that variations in bacterial size may not reflect variations in biomass per cell. PMID- 16347363 TI - Delignification of Wheat Straw by Pleurotus spp. under Mushroom-Growing Conditions. AB - Pleurotus sajor-caju, P. sapidus, P. cornucopiae, and P. ostreatus mushrooms were produced on unsupplemented wheat straw. The yield of mushrooms averaged 3.6% (dry weight basis), with an average 18% straw weight loss. Lignin losses (average, 11%) were lower than cellulose (20%) and hemicellulose (50%) losses. The cellulase digestibility of the residual straw after mushroom harvest was generally lower than that of the original straw. It does not appear feasible to simultaneously produce Pleurotus mushrooms and a highly delignified residue from wheat straw. PMID- 16347364 TI - Application of most-probable-number statistics to direct enumeration of microorganisms. AB - A novel method for rapid determination of total microbial cell numbers was investigated. The method involves the application of most-probable-number estimation statistics to direct microscopic counting of microbial cells by using a particle sizing graticule. Its accuracy and reliability were tested with computer simulations of bacterial cell distributions encountered in ecological studies. Good estimates of cell numbers were obtained when the cell density varied from 3 to 6,000 cells per field, i.e., over 3 orders of magnitude. Low levels of contagion did not markedly influence cell estimates, although high levels, corresponding to discrete scattered microcolonies, did. However, these could be recognized visually. Estimates of cell numbers in Breed smears confirmed its speed and good correlation with the standard quadrat counting technique under real experimental conditions. PMID- 16347365 TI - Regulation of beta-1,4-Endoglucanase Synthesis in Thermomonospora fusca. AB - In Thermomonospora fusca YX, endocellulase synthesis varies over a 100-fold range depending on the carbon source used. This study shows that the variation is caused by two regulatory mechanisms: an induction mechanism that increases the rate of endocellulase synthesis about 20-fold and a growth rate-dependent repression mechanism that changes the rate of synthesis over a 6-fold range in both induced and noninduced cells. In T. fusca, endocellulase synthesis can be induced by cellulose, cellobiose, or cellodextrin. Cellulase is involved in inducer generation from cellulose. Growth rate-dependent repression can be reversed by limiting cultures for carbon, nitrogen, or, to a lesser extent, phosphorus. Further evidence for two separate regulatory mechanisms is provided by the isolation of mutants (CC-1 and CC-2) whose endocellulases are synthesized constitutively but are still sensitive to growth rate-dependent repression. These conclusions about total endocellulase synthesis were extended to the individual endocellulases by showing that three T. fusca endocellulases are coordinately regulated. PMID- 16347366 TI - Reduction of selenate to selenide by sulfate-respiring bacteria: experiments with cell suspensions and estuarine sediments. AB - Washed cell suspensions of Desulfovibrio desulfuricans subsp. aestuarii were capable of reducing nanomolar levels of selenate to selenide as well as sulfate to sulfide. Reduction of these species was inhibited by 1 mM selenate or tungstate. The addition of 1 mM sulfate decreased the reduction of selenate and enhanced the reduction of sulfate. Increasing concentrations of sulfate inhibited rates of selenate reduction but enhanced sulfate reduction rates. Cell suspensions kept in 1 mM selenate were incapable of reducing either selenate or sulfate when the selenate/sulfate ratio was >/=0.02, indicating that irreversible inhibition occurs at high selenate concentrations. Anoxic estuarine sediments having an active flora of sulfate-respiring bacteria were capable of a small amount of selenate reduction when ambient sulfate concentrations were low (<4 mM). These results indicate that sulfate is an inhibitor of the reduction of trace quantities of selenate. Therefore, direct reduction of traces of selenate to selenide by sulfate-respiring bacteria in natural environments is constrained by the ambient concentration of sulfate ions. The significance of this observation with regard to the role sediments play in sequestering selenium is discussed. PMID- 16347367 TI - A differential medium for the enumeration of homofermentative and heterofermentative lactic Acid bacteria. AB - A medium was developed for the differential enumeration of homofermentative and heterofermentative lactic acid bacteria. Essential components of the medium included fructose (14 mM), KH(2)PO(4) (18 mM), bromcresol green (as a pH indicator), and other nutrients to support growth. In agar medium, homofermentative colonies were blue to green, while heterofermentative colonies remained white. A total of 21 Lactobacillus, Pediococcus, Leuconostoc, and Streptococcus species were correctly classified with the medium. PMID- 16347368 TI - Construction of Streptococcus lactis subsp. lactis Strains with a Single Plasmid Associated with Mucoid Phenotype. AB - Lactose-fermenting mucoid (Lac Muc) variants of plasmid-free Streptococcus lactis subsp. lactis MG1614 were obtained by protoplast transformation with total plasmid DNA from MucS. lactis subsp. cremoris ARH87. By using plasmid DNA from these variants for further transformations followed by novobiocininduced plasmid curing, Lac Muc MG1614 strains containing only a single 30-megadalton plasmid could be constructed. This plasmid, designated pVS5, appeared to be associated with the Muc phenotype. PMID- 16347369 TI - Methods for Isolation of Auxotrophic Mutants of Methanobacterium ivanovii and Initial Characterization of Acetate Auxotrophs. AB - To develop a biochemical genetic approach to understanding cell carbon synthesis or metabolic pathways in methanogens, Methanobacterium ivanovii was selected as a model organism for genetic manipulation studies. The organism displayed a colony size of 3 to 6 mm in less than 2 weeks and had a plating efficiency of about 90%, which made it suitable for replica plating. Mutagenesis and selection techniques were developed for selection of acetate auxotrophs. Chemical mutagenesis with ethyl methanesulfonate, followed by enrichment with bacitracin as a selective agent, resulted in stable acetate auxotrophs. M. ivanovii was very sensitive to UV, but UV-induced acetate auxotrophs were unstable and reverted within two to four transfers. The acetate auxotrophs were analyzed in relation to wild type for carbon monoxide dehydrogenase enzyme activity. PMID- 16347370 TI - Production of Pili (Fimbriae) by Pseudomonas fluorescens and Correlation with Attachment to Corn Roots. AB - Pseudomonas fluorescens isolates 13525 and 2-79 were grown in Luria broth and low nutrient medium (LNM). Pililike fibrils were very rarely produced in Luria broth but were abundantly produced in LNM. In LNM the pili were peritrichously distributed and had diameters ranging from 3 to 8 nm. Pili were purified from strain 2-79, and the pilin subunit was found to have a molecular weight of about 34,000. Strain 2-79 produced two colony types on Luria agar, nonmucoidal and mucoidal. Cells in LNM cultures of the nonmucoidal colony type were highly piliated, and cells from the mucoidal type were nearly devoid of pili. The presence of pili on nonmucoidal isolate 2-79 was quantitatively correlated with hydrophobic attachment to polystyrene, hemagglutination, and attachment to corn roots. PMID- 16347371 TI - Sulfate reduction in peat from a new jersey pinelands cedar swamp. AB - Microbial sulfate reduction rates in acidic peat from a New Jersey Pine Barrens cedar swamp in 1986 were similar to sulfate reduction rates in freshwater lake sediments. The rates ranged from a low of 1.0 nmol cm day in February at 7.5- to 10.0-cm depth to 173.4 nmol cm day in July at 5.0- to 7.5-cm depth. The presence of living Sphagnum moss at the surface generally resulted in reduced rates of sulfate reduction. Pore water sulfate concentrations and water table height also apparently affected the sulfate reduction rate. Concentrations of sulfate in pore water were nearly always higher than those in surface water and groundwater, ranging from 26 to 522 muM. The elevated pore water sulfate levels did not result from the evapotranspiratory concentration of infiltrating stream water or groundwater, but probably resulted from oxidation of reduced sulfur compounds, hydrolysis of ester sulfates present in the peat, or both. The total sulfur content of peat that had no living moss at the surface was 164.64 +/- 1.5 and 195.8 +/- 21.7 mumol g (dry weight) for peat collected from 2.5 to 5.0 and 7.5 to 10.0 cm, respectively. Organosulfur compounds accounted for 84 to 88% of the total sulfur that was present in the peat. C-bonded sulfur accounted for 91 to 94% of the organic sulfur, with ester sulfate being only a minor constituent. Reduced inorganic sulfur species in peat from 2.5 to 7.5 cm were dominated by H(2)S-FeS (68%), while pyritic sulfide was the predominant inorganic sulfur species in the peat from depths of 7.5 to 10.0 cm (75%). PMID- 16347372 TI - Iron-Dependent Production of Hydroxamate by Sodium-Dependent Azotobacter chroococcum. AB - The sodium-dependent strain 184 of Azotobacter chroococcum was unable to grow significantly in iron-limited medium, but did produce iron-repressible outer membrane proteins. Siderophores were not produced under these conditions. Citric acid was excreted, but not in response to iron limitation. This strain, however, was able to grow in insoluble mineral iron sources, and under these conditions the cells produced a hydroxamate. Growth on minerals and hydroxamate production was dependent on a low level of freely exchangeable iron. Optimal hydroxamate production was observed with 0.75 muM ferric citrate, and hydroxamate production was repressed by >5 muM iron. Despite this iron requirement, hyroxamate was only formed during internal iron limitation of the cells. Iron-containing cells were able to grow in iron-limited medium but only produced hydroxamate when their iron per-cellular-protein content was low. These results, the spectral changes observed upon Fe addition, and iron-uptake coincident with hydroxamate production suggested that the hydroxamate was a siderophore. PMID- 16347373 TI - Zymomonas mobilis Mutants with an Increased Rate of Alcohol Production. AB - Two new derivatives of Zymomonas mobilis CP4 were isolated from enrichment cultures after 18 months of serial transfers. These new strains were selected for the ability to grow and produce ethanol rapidly on transfer into fresh broth containing ethanol and allyl alcohol. Ethanol production by these strains was examined in batch fermentations under three sets of conditions. Both new derivatives were found to be superior to the parent strain CP4 with respect to the speed and completeness of glucose conversion to ethanol. The best of these, strain YO2, produced 9.5% ethanol (by weight; 11.9% by volume) after 17.4 h compared with 31.8 h for the parent strain CP4. The addition of 1 mM magnesium sulfate improved ethanol production in all three strains. Two factors contributed to the decrease in fermentation time required by the mutants: more rapid growth with minimal lag on subculturing and the retention of higher rates of ethanol production as fermentation proceeded. Alcohol dehydrogenase isozymes were altered in both new strains and no longer catalyzed the oxidation of allyl alcohol into the toxic product acrolein. This loss of allyl alcohol-oxidizing capacity is proposed as a primary factor contributing to increased allyl alcohol resistance, although it is likely that other mutations affecting glycolysis also contribute to the improvement in ethanol production. PMID- 16347374 TI - Detailed characterization and comparison of four lactic streptococcal bacteriophages based on morphology, restriction mapping, DNA homology, and structural protein analysis. AB - Bacteriophages uc1001 and uc1002, which are lytic for Streptococcus cremoris UC501 and UC502, respectively, were characterized in detail. Comparisons were made with a previously characterized phage, P008, which is lytic for Streptococcus lactis subsp. diacetylactis F7/2, and uc3001, which is a lytic phage for S. cremoris UC503. Phages uc1001 and uc1002 had small isometric heads (diameters, 52 and 50 nm, respectively) and noncontractile tails (lengths, 152 and 136 nm, respectively), and uc1002 also had a collar. Both had 30.1 +/- 0.6 kilobase pairs (kbp) of DNA with cross-complementary cohesive ends. Restriction endonuclease maps made with seven endonucleases showed no common fragments. Despite this there was a very high level of homology between uc1001 and uc1002, and results of cross-hybridization experiments showed that the organization of both phage genomes was similar. Heteroduplex analysis confirmed this and quantified the level of homology at 83%. The regions of nonhomology comprised 2.1 , 1.1-, and 1.0-kbp deletion loops and 13 smaller loops and bubbles. The sodium dodecyl sulfate-polyacrylamide gel electrophoretic structural protein profiles were related, with a major band of about 40,000 molecular weight and minor bands of 35,000 and 34,000 molecular weight in common. There were also differences, however, in that uc1001 had a second major band of 68,000 molecular weight and two extra minor bands. Except for the restriction maps, which were strain specific, phages uc1001, uc1002, and P008 were closely related by all the criteria listed above. Their DNAs also showed a very significant bias against the cleavage sites of 9 of 11 restriction endonucleases. Phage uc3001 was unrelated to uc1001, uc1002, or P008 in that it had a prolate head (53 by 39 nm) and a shorter tail (105 nm), contained approximately 22 kbp of DNA, had unrelated cohesive ends, showed no DNA homology with the isometric-headed phages, and displayed a very different structural protein profile. PMID- 16347375 TI - Mating System and Basidiospore Formation in the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium. AB - Prototrophic strains recovered from crosses between auxotrophic strains of the lignin-degrading basidiomycete Phanerochaete chrysosporium were induced to fruit. The progeny of most of these self-crosses were prototrophic, indicating that the nuclei of the original prototroph were wild-type recombinants rather than complementary heterokaryons and that the binucleate basidiospores of this organism are homokaryotic. Various wild-type strains were shown to have multinucleate cells lacking clamp connections and to possess a variable number of sterigmata per basidium. Colonies arising from single conidia of various wild type strains were all capable of producing fruit bodies and basidiospores. In addition, single basidiospores from three wild-type strains all produced fruit bodies and basidiospores. Nonfruiting as well as fruiting isolates were obtained from single basidiospores of five other wild-type strains. Basidiospores from these fruiting isolates always yielded colonies that fruited, again indicating that the spores are homokaryotic. Nonfruiting isolates from the same strain did not produce basidiospores when allowed to form a heterokaryon, implying that these isolates do not represent mating types. All this evidence indicates that P. chrysosporium has a primary homothallic mating system. In addition to fruiting and nonfruiting phenotypes, basidiospores from strain OGC101, a derivative of ME 446, gave rise to colonies which did not grow on cellulose (Cel). The fruiting, nonfruiting, and Cel phenotypes differed from each other and from the parental wild-type strain in a variety of characteristics, including growth, conidiation, and evolution of CO(2) from C-side chain-labeled lignin, indicating that strain OCG101 is a heterokaryon. PMID- 16347376 TI - Expression of a Thermomonospora fusca Cellulase Gene in Streptomyces lividans and Bacillus subtilis. AB - A cellulase gene from Thermomonospora fusca coding for endocellulase E(5) was introduced into Streptomyces lividans by using shuttle plasmids that can replicate in either S. lividans or Escherichia coli. Plasmid DNA isolated from E. coli was used to transform S. lividans, selecting for thiostrepton resistance. The transformants expressed and excreted the endocellulase, but the ability to produce the endocellulase was unstable. This instability was shown to result from deletion of the endocellulase gene from the plasmid. Plasmid DNA prepared from a culture in which plasmid modification had occurred was used to transform E. coli, selecting for Amp cells, and all of the transformants were cellulase positive, showing that pBR322 and T. fusca DNA were deleted together. When a plasmid was constructed containing only T. fusca DNA in plasmid pIJ702, the transformants were more stable, and the level of endocellulase activity produced in the culture supernatant after growth on 0.2% glucose was close to the level produced by T. fusca cultures grown on 0.2% cellulose. About 50% of the total protein in the culture supernatant of the S. lividans transformant was endocellulase E(5). The enzyme produced by the S. lividans transformant was identical to pure T. fusca E(5) in its electrophoretic mobility and was completely inhibited by antiserum to E(5). Shuttle plasmids containing the E(5) gene that could replicate in Bacillus subtilis and E. coli were also constructed and used to transform B. subtilis. Again there was extensive deletion of the plasmid DNA during transformation and growth in B. subtilis. There was no evidence of E(5) activity, even in those B. subtilis transformants that retained the E(5) gene. PMID- 16347377 TI - Efficient Implementation of Consecutive Reactions by Peptidases at the Periphery of the Streptococcus cremoris Membrane. AB - Activities detectable in Streptococcus cremoris with the chymotrypsin substrate N glutaryl-l-phenylalanine-4-nitroanilide and formerly designated endopeptidases P37 and P50 (F. A. Exterkate, Appl. Environ. Microbiol. 47:177-183, 1984) are both coupled peptidase reactions. These coupled reactions involve a membrane bound, restricted l-alpha-glutamyl aminopeptidase which is responsible for the initial release of the glutaryl moiety. The subsequent reaction is catalyzed by either a so-called low-temperature or a high-temperature phenylalanyl aminopeptidase activity, both located at the outside surface of the membrane. Altered microenvironmental conditions created by the membrane-perturbing action of n-butanol or obtained by solubilization resulted in the removal of a restriction on the activity of l-alpha-glutamyl aminopeptidase and in a less efficient functioning of the coupled reactions; a long transient phase occurred before the steady state was reached. The results suggest that the in situ spatial organization is conducive to an efficient attuning of at least three peptidases which are located at the outer membrane surface and in the membrane. The possibility that peptidases in these locations exist as a cluster with physiological significance is discussed in relation to growth of S. cremoris in milk. PMID- 16347379 TI - Induction and Characterization of Penicillium caseicolum Mutants Resistant to Ergosterol Biosynthesis Inhibitors. AB - The isolation of Penicillium caseicolum mutants resistant to different fungicides which inhibit ergosterol biosynthesis is reported. Mutational frequencies for resistance were high (3 x 10 to 3 x 10). The levels of resistance toward the inhibitors of sterol C-14 demethylation were always low (<10), whereas high values were obtained with mutants resistant to inhibitors of sterol Delta14 reduction or Delta8-->Delta7 isomerization, or both. Generally, there was a positive cross-resistance between fungicides showing the same biochemical mode of action but not between compounds of two different groups. Mycelial growth rate and sporulation were tested; several mutants were not affected for these characteristics. We conclude that resistance to ergosterol biosynthesis inhibitors may be used as a good marker for genetic studies through protoplast fusion. PMID- 16347378 TI - Variation in Quantitative Requirements for Na for Transport of Metabolizable Compounds by the Marine Bacteria Alteromonas haloplanktis 214 and Vibrio fischeri. AB - The rates of uptake by Alteromonas haloplanktis of 19 metabolizable compounds and by V. fischeri of 16 of 17 metabolizable compounds were negligible in the absence of added alkali-metal cations but rapid in the presence of Na. Only d-glucose uptake by V. fischeri occurred at a reasonable rate in the absence of alkali metal cations, although the rate was further increased by added Na, K, or Li. Quantitative requirements for Na for the uptake of 11 metabolites by A. haloplanktis and of 6 metabolites by V. fischeri and the characteristics of the Na response at constant osmotic pressure varied with each metabolite and were different from the Na effects on the energy sources used. Li stimulated transport of some metabolites in the presence of suboptimal Na concentrations and for a few replaced Na for transport but functioned less effectively. K had a small capacity to stimulate lysine transport. The rate of transport of most of the compounds increased to a maximum at 50 to 300 mM Na, depending on the metabolite, and then decreased as the Na concentration was further increased. For a few metabolites, the rate of transport continued to increase in a biphasic manner as the Na concentration was increased to 500 mM. Concentrations of choline chloride equimolar to inhibitory concentrations of NaCl were either not inhibitory or appreciably less inhibitory than those of NaCl. All metabolites examined accumulated inside the cells against a gradient of unchanged metabolite in the presence of Na, even though some were very rapidly metabolized. The transport of l-alanine, succinate, and d-galactose into A. haloplanktis and of l-alanine and succinate into V. fischeri was inhibited essentially completely by the uncoupler 3,5,3',4'-tetrachlorosalicylanilide. Glucose uptake by V. fischeri was inhibited partially by 3,5,3',4'-tetrachlorosalicylanilide and also by arsenate and iodoacetate. PMID- 16347380 TI - Induction and Characterization of Artificial Diploids from the Haploid Yeast Torulaspora delbrueckii. AB - The yeast Torulaspora delbrueckii, which propagates as a haploid, was made into a diploid by treatment with dimethyl sulfoxide (DMSO) on the regeneration of protoplasts. The diploid state was stably inherited; the cell volume was three times that of the parent strain and the cellular DNA content was two times that of the parental strain. No essential difference was found between diploids induced by DMSO and those formed through intraspecific protoplast fusion. The diploid strains sporulated fairly well, with their cells converting directly into asci. Random spore analysis revealed that diploids induced through protoplast fusion gave rise to auxotrophic segregants (haploids) with the parental genetic marker or to segregants formed by recombination, while diploids induced by DMSO from a doubly auxotrophic parent gave rise to no recombinant, indicating that it was chromosomally homoallelic in nature. The magnesium level in the protoplast regeneration medium was found to be an important factor for inducing diploid formation. At 0.2 mM magnesium diploids appeared even in the absence of DMSO, while at 2 mM magnesium diploids never appeared unless DMSO was added to the regeneration medium. Evidence is provided that the diploids induced by DMSO or a low magnesium level are due to direct diploidization but not protoplast fusion. UV light irradiation of intact cells (without protoplasts), 10% of which survived, also produced diploids among this surviving population. From these results we conclude that the perturbation of protoplast regeneration or of cell division by the treatments mentioned above somehow induced direct diploidization of T. delbrueckii. PMID- 16347381 TI - Industrial Application of Artificially Induced Diploid Strains of Torulaspora delbrueckii. AB - Diploid strains of Torulaspora delbrueckii were tested for industrial application. Because the cell volume of the diploid strain was three times as large as that of the parental haploid strain, collection and subsequent dehydration to make compressed yeast cakes were greatly improved with the diploid YL3. The time required for dehydration of the diploid strain was shortened to 1/2.5 that of the parent strain under conventional conditions. Moreover, for the diploid cells frequent filter changes for dehydration were not required, which was the case with parental cells. Fermentation activity and tolerance to freeze thawing in dough were succesfully inherited by the diploid strains. The diploid YL3 showed nearly the same activity as the diploid F31 in bread making. However, the endurance period of yeast cakes when stored at 30 degrees C without softening to lead to liquefaction was much longer in YL3 (199 h) than in F31 (132 h). This superiority was ascribed to the fact that YL3 was induced through direct diploidization and had no genetic defect on chromosomes because the wild-type strain was employed as the parent, whereas F31 was obtained through protoplast fusion from two auxotrophic mutants and carried at least two mutagenized genes that were masked by heterolallelism. PMID- 16347382 TI - Relation to copper of N-1, a nonobligate bacterial predator. AB - Nonobligate bacterial predator strain N-1 was highly resistant to copper. In fact, it required more than minimal amounts of copper to initiate growth, but not for growth that followed growth initiation. Strain N-1 made a peptide growth initiation factor (GIF) to marshal copper from its environment for growth initiation. Production of this GIF occurred before the onset of growth initiation, but production was shut down if excess copper was present. At high copper levels, the time required for onset of growth initiation was directly related to the amount of copper that was present. At low copper levels, a similar graded response occurred for increments of added GIF. Agromyces ramosus is a predator in its own right, but it also is a prey species for strain N-1. A. ramosus was found to be very sensitive to copper and to the copper GIF produced by N-1. It is possible that the copper GIF is the means used by N-1 to kill A. ramosus. PMID- 16347383 TI - Diversity among Opine-Utilizing Bacteria: Identification of Coryneform Isolates. AB - Bacteria were isolated from soil and crown gall tumors by selection in minimal medium with an opine, such as succinamopine or mannopine, as the sole carbon source. The isolates were characterized for the pattern of opine utilization and identified. They were classified as mannityl opine or imino diacid utilizers and exhibited specificity of utilization similar to that described previously for Agrobacterium species. A minority of isolates were gram negative and were identified as Agrobacterium or Pseudomonas species; most were gram positive and belonged to the coryneform group. These results indicate that any specific effect of opines on the ecology of Agrobacterium tumefaciens is modulated by activities of other types of soil- and plant-associated bacteria. PMID- 16347384 TI - Rapid assay for microbially reducible ferric iron in aquatic sediments. AB - The availability of ferric iron for microbial reduction as directly determined by the activity of iron-reducing organisms was compared with its availability as determined by a newly developed chemical assay for microbially reducible iron. The chemical assay was based on the reduction of poorly crystalline ferric iron by hydroxylamine under acidic conditions. There was a strong correlation between the extent to which hydroxylamine could reduce various synthetic ferric iron forms and the susceptibility of the iron to microbial reduction in an enrichment culture of iron-reducing organisms. When sediments that contained hydroxylamine reducible ferric iron were incubated under anaerobic conditions, ferrous iron accumulated as the concentration of hydroxylamine-reducible ferric iron declined over time. Ferrous iron production stopped as soon as the hydroxylamine-reducible ferric iron was depleted. In anaerobic incubations of reduced sediments that did not contain hydroxylamine-reducible ferric iron, there was no microbial iron reduction, even though the sediments contained high concentrations of oxalate extractable ferric iron. A correspondence between the presence of hydroxylamine reducible ferric iron and the extent of ferric iron reduction in anaerobic incubations was observed in sediments from an aquifer and in fresh- and brackish water sediments from the Potomac River estuary. The assay is a significant improvement over previously described procedures for the determination of hydroxylamine-reducible ferric iron because it provides a correction for the high concentrations of solid ferrous iron which may also be extracted from sediments with acid. This is a rapid, simple technique to determine whether ferric iron is available for microbial reduction. PMID- 16347385 TI - Ultrastructure of Thiothrix spp. and "Type 021N" Bacteria. AB - The ultrastructural features of two groups of filamentous sulfur bacteria, Thiothrix spp. and an unnamed organism designated "type 021N," were examined by transmission electron microscopy. Negative staining of whole cells and filaments with uranyl acetate revealed the presence of tufts of fimbriae located at the ends of individual gonidia of Thiothrix sp. strain A1 and "type 021N" strain N7. Holdfast material present at the center of mature rosettes was observed in thin sections stained with ruthenium red. A clearly defined sheath enveloped the trichomes of two of three Thiothrix strains but was absent from "type 021N" filaments. The outer cell wall appeared more complex in "type 021N" strains than in Thiothrix isolates. Bulbs or clusters of irregularly shaped cells, often present in filaments of "type 021N" bacteria, appeared to result from crosswalls which formed at angles oblique to the filament axis. The multicellular nature of these sulfur bacteria was apparent in that only the cytoplasmic membrane and peptidoglycan layer of the cell wall were involved in the septation process. Sulfur inclusions which developed in the presence of sodium thiosulfate were enclosed by a single-layered envelope and located within invaginations of the cytoplasmic membrane. PMID- 16347386 TI - Cloning of a Streptococcus lactis subsp. lactis Chromosomal Fragment Associated with the Ability To Grow in Milk. AB - A chromosomal fragment of 6.7 megadaltons (MDa), apparently containing the genes for milk protein utilization by Streptococcus lactis subsp. lactis SSL135, was cloned in S. lactis subsp. lactis MG1614, a proteinase-negative strain. For the cloning, the chromosomal DNA of SSL135 was cleaved with restriction enzyme BamHI and the resulting fragments were ligated to the single BclI site of pVS2, a 3.3 MDa chloramphenicol-erythromycin double-resistance plasmid constructed in this laboratory. S. lactis subsp. lactis MG1614 was transformed by using this ligation mixture and selecting for chloramphenicol resistance and growth in citrated milk medium. One clone containing a 10.0-MDa plasmid, subsequently designated as pVS6, was chosen for further studies. Despite the lack of homology with previously characterized proteinase genes of lactic streptococci, the cloned insert consistently conveyed the ability to grow in milk to proteinase-negative recipients in repeated transformation experiments. The genetic evidence suggests that the main part of the gene(s) for the proposed proteinase activity is located within a 3.8-MDa BglII fragment of the clone. PMID- 16347387 TI - Effects of pH, Temperature, and Nutrients on Propionate Degradation by a Methanogenic Enrichment Culture. AB - Enrichment cultures were used to determine the conditions promoting fastest methanogenic propionate degradation and growth by adapting the cultures to various physical and chemical conditions and measuring the specific growth rate. We found that the fastest growth of propionate oxidizers occurred at pH 6.8 to 8.5 and 32 to 45 degrees C. Acetate-degrading populations showed narrower optima for fastest growth (pH 6.8 to 7.2 and 37 to 43 degrees C). Enrichment cultures grew as well in minimal medium as in complex medium, although individual microbial populations appeared to require growth factors which could be met by cross-feeding. PMID- 16347388 TI - Isozyme Variation among 40 Frankia Strains. AB - Forty Frankia strains belonging to the Alnus and Elaeagnus host specificity groups and isolated from various plant species from different geographical areas were characterized by the electrophoretic separation of isozymes of eight enzymes. All the enzyme systems that were investigated showed large variation. Diaphorases and esterases gave multiple band patterns and confirmed the identification of specific Frankia strains. Less variability was observed with enzymes such as phosphoglucose isomerase, leucine aminopeptidase, and malate dehydrogenase, which allowed for the delineation of larger groups of Frankia strains. Cluster analysis, based on the pair-wise similarity coefficients calculated between strains, delineated three large, dissimilar groups of Frankia strains, although each of these groups contained a large amount of heterogeneity. However, numerous Frankia strains, mainly from the Alnus host specificity group, demonstrated a perfect homology for all the enzymes tested. PMID- 16347389 TI - Production of ethane, ethylene, and acetylene from halogenated hydrocarbons by methanogenic bacteria. AB - Several methanogenic bacteria were shown to produce ethane, ethylene, and acetylene when exposed to the halogenated hydrocarbons bromoethane, dibromo- or dichloroethane, and 1,2-dibromoethylene, respectively. They also produced ethylene when exposed to the coenzyme M analog and specific methanogenic inhibitor bromoethanesulfonic acid. The production of these gases from halogenated hydrocarbons has a variety of implications concerning microbial ecology, agriculture, and toxic waste treatment. All halogenated aliphatic compounds tested were inhibitory to methanogens. Methanococcus thermolithotrophicus, Methanococcus deltae, and Methanobacterium thermoautotrophicum DeltaH and Marburg were completely inhibited by 7 muM 1,2 dibromoethane and, to various degrees, by 51 to 1,084 muM 1,2-dichloroethane, 1,2 dibromoethylene, 1,2-dichloroethylene, and trichloroethylene. In general, the brominated compounds were more inhibitory. The two Methanococcus species were fully inhibited by 1 muM bromoethanesulfonic acid, whereas both Methanobacterium strains were only partly inhibited by 2,124 muM. Coenzyme M protected cells from bromoethanesulfonic acid but not from any of the other inhibitors. PMID- 16347390 TI - Split-Root Assays Using Trifolium subterraneum Show that Rhizobium Infection Induces a Systemic Response That Can Inhibit Nodulation of Another Invasive Rhizobium Strain. AB - Subterranean clover plants possessing two equally infectible and robust lateral root systems ("split roots") were used in conjunction with several specific mutant strains (derived from Rhizobium trifolii ANU843) to investigate a systemic plant response induced by infective Rhizobium strains. This plant response controls and inhibits subsequent nodulation on the plant. When strain ANU843 was inoculated onto both root systems simultaneously or 24, 48, 72, or 96 h apart, an inhibitory response occurred which retarded nodulation on the root exposed to the delayed inoculum but only when the delay period between inocula was greater than 24 h. Equal numbers of nodules were generated on both roots when ANU843 was inoculated simultaneously or 24 h apart. The ability to infect subterranean clover plants was required to initiate the plant inhibitory response since preexposure of one root system to non-nodulating strains did not retard the ability of the wild-type strain to nodulate the opposing root system (even when the delay period was 96 h). Moreover, the use of specific Tn5-induced mutants subtly impaired in their ability to nodulate demonstrated that the plant could effectively and rapidly discriminate between infections initiated by either the parent or the mutant strains. When inoculated alone onto clover plants, these mutant strains were able to infect the most susceptible plant cells at the time of inoculation and induce nitrogen-fixing nodules. However, the separate but simultaneous inoculation on opposing root systems of the parent and the mutant strains resulted in the almost complete inhibition of the nodulation ability of the mutant strains. We concluded that the mutants were affected in their competitive ability, and this finding was reflected by poor nodule occupancy when the mutants were coinoculated with the parent strain onto a single root system. Thus the split-root system may form the basis of a simple screening method for the ranking of competitiveness of various rhizobia on small seeded legumes. PMID- 16347391 TI - Importance of Hydrogen Sulfide, Thiosulfate, and Methylmercaptan for Growth of Thiobacilli during Simulation of Concrete Corrosion. AB - Biogenic sulfuric acid corrosion of concrete surfaces caused by thiobacilli was reproduced in simulation experiments. At 9 months after inoculation with thiobacilli, concrete blocks were severely corroded. The sulfur compounds hydrogen sulfide, thiosulfate, and methylmercaptan were tested for their corrosive action. With hydrogen sulfide, severe corrosion was noted. The flora was dominated by Thiobacillus thiooxidans. Thiosulfate led to medium corrosion and a dominance of Thiobacillus neapolitanus and Thiobacillus intermedius. Methylmercaptan resulted in negligible corrosion. A flora of heterotrophs and fungi grew on the blocks. This result implies that methylmercaptan cannot be degraded by thiobacilli. PMID- 16347392 TI - Thermostable amylolytic enzymes from a new clostridium isolate. AB - A new Clostridium strain was isolated on starch at 60 degrees C. Starch, pullulan, maltotriose, and maltose induced the synthesis of alpha-amylase and pullulanase, while glucose, ribose, fructose, and lactose did not. The formation of the amylolytic enzymes was dependent on growth and occurred predominantly in the exponential phase. The enzymes were largely cell bound during growth of the organism with 0.5% starch, but an increase of the starch concentration in the growth medium was accompanied by the excretion of alpha-amylase and pullulanase into the culture broth; but also by a decrease of total activity. alpha-Amylase, pullulanase, and alpha-glucosidase were active in a broad temperature range (40 to 85 degrees C) and displayed temperature optima for activity at 60 to 70 degrees C. During incubation with starch under aerobic conditions at 75 degrees C for 2 h, the activity of both enzymes decreased to only 90 or 80%. The apparent K(m) values of alpha-amylase, pullulanase, and alpha-glucosidase for their corresponding substrates, starch, pullulan, and maltose were 0.35 mg/ml, 0.63 mg/ml, and 25 mM, respectively. PMID- 16347393 TI - Production of Thermostable alpha-Amylase, Pullulanase, and alpha-Glucosidase in Continuous Culture by a New Clostridium Isolate. AB - The production of alpha-amylase, pullulanase, and alpha-glucosidase and the formation of fermentation products by the newly isolated thermophilic Clostridium sp. strain EM1 were investigated in continuous culture with a defined medium and an incubation temperature of 60 degrees C. Enzyme production and excretion were greatly influenced by the dilution rate and the pH of the medium. The optimal values for the formation of starch-hydrolyzing enzymes were a pH of 5.9 and a dilution rate of 0.075 to 0.10 per h. Increase of the dilution rate from 0.1 to 0.3 per h caused a drastic drop in enzyme production. The ethanol concentration and optical density of the culture, however, remained almost constant. Growth limitation in the chemostat with 1% (wt/vol) starch was found optimal for enzyme production. Under these conditions 2,800 U of pullulanase per liter and 1,450 U of alpha-amylase per liter were produced; the amounts excreted were 70 and 55%, respectively. PMID- 16347394 TI - Free-living dinitrogen-fixing bacteria isolated from petroleum refinery oily sludge. AB - Dinitrogen-fixing activity (acetylene reduction and N(2) fixation) was found in an oily sludge originating from a petroleum refinery. Two representative dinitrogen-fixing bacterial strains were isolated from this oily waste. Their nitrogenase activity was effective when they were cultivated on sterilized sludge or simple carbon substrates (organic acid salts, sugars). Using the classical methods, these strains could not be unambiguously related to other diazotrophic taxa. The landfarming process is widely used for oily sludge disposal; this study shows that oily sludges are more than a simple carbon input into the soil but that they must also be considered as real sources of dinitrogen-fixing and probably degradative microorganisms. PMID- 16347395 TI - Purification and Properties of an Extracellular Protease Produced by the Entomopathogenic Fungus Beauveria bassiana. AB - Beauveria bassiana GK2016 grown in a medium with gelatin as the sole carbon and nitrogen source produced an extracellular protease. The protease production was highest when the fungus was grown on a semiliquid medium and was purified about 18-fold, with a recovery of 21%. The protease molecular weight was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be about 35,000. It had an optimum activity at pH 8.5 and 37 degrees C and was rapidly inactivated at 50 degrees C. Its enzymatic activity was that of an endopeptidase which hydrolyzed elastin, casein, and gelatin but was much less active on bovine serum albumin and collagen. No trypsinlike activity was detected on N-alpha-benzoyl-dl arginine-p-nitroanilide. It was, however, inhibited by phenylmethylsulfonyl fluoride, indicating that a serine residue is present in the active site. The protease was unaffected by metal-chelating agents, sulfhydryl reagents, trypsin inhibitor, and chymotrypsin inhibitor. PMID- 16347396 TI - Sulfur Reduction by the Extremely Thermophilic Archaebacterium Pyrodictium occultum. AB - The relationship between growth and biological sulfur reduction for the extremely thermophilic archaebacterium Pyrodictium occultum was studied over a temperature range of 98 to 105 degrees C. The addition of yeast extract (0.2 g/liter) to the medium was found to increase hydrogen sulfide production significantly, especially at higher temperatures. Sulfide production in uninoculated controls with and without yeast extract was noticeable but substantially below the levels observed in samples containing the microorganism. PMID- 16347397 TI - Optimization of Protoplast Formation and Regeneration in Leuconostoc mesenteroides. AB - Conditions are reported for efficient protoplast formation and regeneration in four strains of Leuconostoc mesenteroides. Protoplasts were produced from each strain at frequencies greater than 99%, although the rate of their production was variable from strain to strain. Bovine serum albumin and Mg were required for maximal regeneration, while the presence of Ca was inhibitory. Regeneration frequencies of 16% could be obtained with strain ATCC 10830. This frequency was four- to eightfold higher than the frequencies of the other strains examined. PMID- 16347398 TI - Sporulation of Clostridium cylindrosporum on a Defined, Low-Manganese Medium. AB - Clostridium cylindrosporum HC-1 grew and sporulated well on a defined medium. This is the first demonstration of sporulation of a purinolytic clostridium on a defined medium; manganese levels were below those considered essential for sporulation of most Bacillus species. Sporulation appeared to be initiated before exhaustion of the purine substrate. PMID- 16347399 TI - Effects of Calcium, Magnesium, pH, and Extent of Growth on the Morphology of Methanosarcina mazei S-6. AB - Methanosarcina mazei S-6 grew faster and its morphology changed to individual coccoid cells in medium with elevated concentrations of divalent cations and a large amount of catabolic substrate. PMID- 16347400 TI - Identification of lotus rhizobia by direct DNA hybridization of crushed root nodules. AB - Hybridization of crushed Lotus pedunculatus root nodules with P-labeled total genomic DNA probes was used to identify Rhizobium loti and Bradyrhizobium sp. (Lotus rhizobia). Probes always hybridized with homologous target DNA and frequently with DNAs of other strains from the same genus. Intergeneric hybridization did not occur. Results were comparable to those from colony hybridization. PMID- 16347401 TI - Methods To Alter the Recovery and Nodule Location of Bradyrhizobium japonicum Inoculant Strains on Field-Grown Soybeans. AB - Three strains of Bradyrhizobium japonicum, I17, 110, and 61A76, were evaluated for their ability to form nodules on field-grown soybeans in soil with a highly competitive indigenous B. japonicum population. The predominant indigenous strain, 0336, in the field site used was unlike the more common isolates from Midwestern soils which belong to the 123 or 138 serogroups. This strain persisted in the soil for at least 30 years without any soybean crops. The three inoculant strains differed in their ability to compete with indigenous strains for nodule formation. Four different inoculation treatments were tested in three adjacent fields. When the amount of inoculum was increased, a higher proportion of nodules contained the inoculant strain. The most competitive inoculant strain was I17, a recent field isolate. Strain 61A76 was better than 110. There was no difference in recovery of the inoculant strains on the Hodgson or Corsoy soybean cultivars, nor was there a difference in recovery of the inoculant strains during the growing season. The vertical distribution of nodules containing the inoculant strains was affected by the method of adding the inoculant to the soil. Inoculant added to the seed furrow produced nodules mainly in the top region of the soybean root. Inoculant tilled into the soil produced nodules primarily in the bottom part of the root. The nodules that were produced in the bottom part of the root are younger and may contribute significant amounts of fixed nitrogen to the soybean during seed formation. PMID- 16347402 TI - Preferential Utilization of Cellobiose by Thermomonospora curvata. AB - Thermomonospora curvata was cultivated on mineral salts medium containing glucose and cellobiose under conditions that increasingly favored the uptake of glucose. In each case cellobiose was utilized in preference to glucose and induced beta glucosidase and endoglucanase activity. [C]glucose metabolism studies indicated that cellobiose was not cleaved by extracellular beta-glucosidase and transported as glucose. No evidence of cellobiose phosphorylase or a cellobiose-specific phosphoenolpyruvate-phosphotransferase system was observed. PMID- 16347403 TI - Seasonal and spatial distribution of extracellular enzymatic activities and microbial incorporation of dissolved organic substrates in marine sediments. AB - Seasonal and spatial distributions of extracellular enzymatic activities and microbial incorporations of dissolved organic substrates were followed in sediments of the brackish water Kiel Bight (Baltic Sea, Federal Republic of Germany). Enzymatic hydrolysis of polymeric organic compounds was determined by means of fluorogenic substrates (4-methylumbelliferyl-beta-d-glucoside, l-leucine 4-methylcoumarinyl-7-amide hydrochloride); incorporation of dissolved organic substrates into microbial biomass was measured by using tritiated substances (acetate, leucine, and thymidine). Based on a recently developed core injection technique, substrates were injected in microliter portions into undisturbed sediment cores. Enzymatic and incorporation activities underwent strong seasonal variations related to the enrichment of organic material in the sediment surface following sedimentation events. The input of the phytoplankton bloom during autumn caused stimulation of both enzymatic hydrolysis of polymeric organic compounds and microbial incorporation of dissolved organic substrates. Following input by spring phytoplankton bloom, mainly incorporation activities were stimulated. In late spring the development of the benthic fauna obviously greatly influenced microbial activities. During summer individual periods of high microbial activities were observed which might be traced back to short-term sedimentation events. The high microbial incorporation of leucine and thymidine during winter demonstrated that the nutrient supply rather than temperature is the dominating factor determining microbial production. Stimulation of microbial activities arose from the sediment surface and spread out relatively quickly into deeper horizons. Generally, the sediments were characterized by distinct patterns of interrelationships between the individual parameters of microbial activities measured. PMID- 16347404 TI - Serological Relatedness of Rhizobium fredii to Other Rhizobia and to the Bradyrhizobia. AB - Several isolates of Rhizobium fredii were examined for their serological relatedness to each other, to Bradyrhizobium japonicum, and to other fast- and slow-growing rhizobia. Immunofluorescence, agglutination, and immunodiffusion analyses indicated that R. fredii contains at least three separate somatic serogroups, USDA 192, USDA 194, and USDA 205. There was no cross-reaction between any of the R. fredii isolates and 13 of the 14 B. japonicum somatic serogroups tested. Cross-reactions were obtained with antisera from R. fredii and serogroup 122 of B. japonicum, Rhizobium meliloti, and several fast-growing Rhizobium spp. for Leucaena, Sesbania, and Lablab species. The serological relationship between R. fredii and R. meliloti was examined in more detail, and of 23 R. meliloti strains examined, 8 shared somatic antigens with the type strains from all three R. fredii serogroups. The serological relatedness of R. fredii to B. japonicum and R. meliloti appears to be unique since the strains are known to be biochemically and genetically diverse. PMID- 16347405 TI - High-Efficiency Conversion of Pyruvate to Acetoin by Lactobacillus plantarum during pH-Controlled and Fed-Batch Fermentations. AB - The influence of pH on the type and concentration of metabolites produced from pyruvate by Lactobacillus plantarum ATCC 8014 was examined in pH-controlled fermentors at pH values of 4.5 to 6.5. Specific growth rates, cell dry weights, and diacetyl concentrations were highest at pH 5.5, with values of 0.78 h, 190 mg/liter, and 1.2 mM, respectively. While the conversion efficiency (millimoles of acetoin formed per millimoles of pyruvate utilized) was highest (94.6%) at pH 4.5, acetoin levels were similar (20 mM) between pH 4.5 and 5.5. Feeding stationary-phase cells exogenous pyruvate increased acetoin levels to 78 mM. PMID- 16347406 TI - Isolation and Characterization of a Catabolite Repression-Insensitive Mutant of a Methanol Yeast, Candida boidinii A5, Producing Alcohol Oxidase in Glucose Containing Medium. AB - Mutants exhibiting alcohol oxidase (EC 1.1.3.13) activity when grown on glucose in the presence of methanol were found among 2-deoxyglucose-resistant mutants derived from a methanol yeast, Candida boidinii A5. One of these mutants, strain ADU-15, showed the highest alcohol oxidase activity in glucose-containing medium. The growth characteristics and also the induction and degradation of alcohol oxidase were compared with the parent strain and mutant strain ADU-15. In the parent strain, initiation of alcohol oxidase synthesis was delayed by the addition of 0.5% glucose to the methanol medium, whereas it was not delayed in mutant strain ADU-15. This showed that alcohol oxidase underwent repression by glucose. On the other hand, degradation of alcohol oxidase after transfer of the cells from methanol to glucose medium (catabolite inactivation) was observed to proceed at similar rates in parent and mutant strains. The results of immunochemical titration experiments suggest that catabolite inactivation of alcohol oxidase is coupled with a quantitative change in the enzyme. Mutant strain ADU-15 was proved to be a catabolite repression-insensitive mutant and to produce alcohol oxidase in the presence of glucose. However, it was not an overproducer of alcohol oxidase and, in both the parent and mutant strains, alcohol oxidase was completely repressed by ethanol. PMID- 16347407 TI - Phototaxis in the Marine Fungus Rhizophydium littoreum. AB - Phototaxis appears to be a factor that influences the dispersal of zoospores of the marine fungus Rhizophydium littoreum. By using a quantitative method to study phototaxis, zoospores were found to be positively phototactic toward blue wavelengths of light. White light in the range of 20 to 6,000 microeinsteins m s and blue light in the range of 1 to 300 microeinsteins m s gave positive responses in the laboratory. Results of field studies confirmed the ability of zoospores to respond to light under natural conditions. Phototaxis may be an important adaptive mechanism which influences vertical migration of zoospores in the photic zone where plant products are available as nutrients. PMID- 16347408 TI - Incorporation of Exogenous Purines and Pyrimidines by Methanococcus voltae and Isolation of Analog-Resistant Mutants. AB - Methanococcus voltae incorporated exogenous adenine, guanine, hypoxanthine, and uracil, but not thymine. Growth of M. voltae was also sensitive to purine and pyrimidine analogs. Of the 20 analogs tested, 12 were inhibitory at 1 mg/ml. The most effective inhibitors were purine analogs with endocyclic substitutions. Nucleoside analogs and analogs with exocyclic substitutions or additions were less effective. Four purine analogs, 8-aza-2,6-diaminopurine, 8-azaguanine, 8 azahypoxanthine, and 6-mercaptopurine and one pyrimidine analog, 6-azauracil, were especially toxic. The MICs were 20, 0.5, 2.0, 80, and 10 mug/ml, respectively. Spontaneous resistance mutants were isolated for these five analogs. The MICs for these mutants were 20.5, 8.2, >65, >41, and 20.5 mg/ml, respectively. These concentrations far exceeded the solubilities of the analogs and represented an increase in resistance of at least three orders of magnitude. In addition to demonstrating cross resistance to several of the analogs, four of these mutants lost the ability to incorporate exogenous bases. These appeared to be mutations in the salvage pathways for purines and pyrimidines. In contrast, the mutant resistant to 6-mercaptopurine was not defective in purine uptake. Instead, it degraded 6-mercaptopurine. In the presence or absence of high concentrations of the analogs, the growth rates of the resistant mutants were no less than one-half of the growth rate of the wild type in the absence of the analog. The high level of resistance and rapid growth are very desirable properties for the application of the mutants in genetic experiments. PMID- 16347409 TI - Auxin production by plant-pathogenic pseudomonads and xanthomonads. AB - Pathogenic strains of Xanthomonas campestris pv. glycines which cause hypertrophy of leaf cells of susceptible soybean cultivars and nonpathogenic strains which do not cause hypertrophy were compared for their ability to produce indole compounds, including the plant hormone indole-3-acetic acid (IAA) in liquid media with or without supplementation with l-tryptophan. Several additional strains of plant-pathogenic xanthomonads and pseudomonads were also tested for IAA production to determine whether in vitro production of IAA is related to the ability to induce hypertrophic growth of host tissues. Indoles present in culture filtrates were identified by thin-layer chromatography, high-performance liquid chromatography, UV spectroscopy, mass spectroscopy, and gas chromatography-mass spectrometry and were quantitated by high-performance liquid chromatography. All strains examined produced IAA when liquid media were supplemented with l tryptophan. The highest levels of IAA were found in culture filtrates from the common bean pathogen Pseudomonas syringae pv. syringae, and this was the only bacterium tested which produced IAA without addition of tryptophan to the medium. Additional indoles identified in culture filtrates of the various strains included indole-3-lactic acid, indole-3-aldehyde, indole-3-acetamide, and N acetyltryptophan. Pseudomonads and xanthomonads could be distinguished by the presence of N-acetyltryptophan, which was found only in xanthomonad culture filtrates. PMID- 16347410 TI - Interactions of Lactobacillus bulgaricus Temperate Bacteriophage 0448 with Host Strains. AB - Lactobacillus bulgaricus LT4(0448) is a lysogenic strain from which a temperate bacteriophage can be induced by mitomycin C or UV irradiation. Lactobacillus lactis CNRZ 326 is an indicator strain for the temperate phage 0448, but this strain lyses only in the presence of Ca ions. A resistant culture developed secondarily after phage lysis and grew normally in MRS broth but again lysed abruptly if Ca ions were added after two or three transfers. This behavior of the secondary culture and its subcultures is explained by a heterogeneous and fluctuating bacterial population, including clones identical to L. lactis 326, which were sensitive to 0448 and which formed rough colonies, as does the indicator. The proportion of these clones increased in the course of transfers in MRS, explaining lysis when Ca was added. The population also included clones which formed smooth colonies (S clones). SI clones, which could not be induced by mitomycin C, were the major type in the initial culture, although they were sensitive to temperate phage 0448. The SI population then decreased and was gradually replaced by SII clones, inducible by mitomycin C and resistant to 0448. These SII clones were lysogenized clones, 326(0448), whose stability was confirmed by growth in the presence of an antiphage serum. When L. bulgaricus LT4(0448) was treated with mitomycin C, several cured LT4 clones were obtained that were related to the clones of the indicator L. lactis 326; they formed rough colonies. They also became sensitive to lytic phages or temperate phages active against L. lactis 326 and insensitive to lytic phages which lysed L. bulgaricus LT4(0448). This suggests that phage 0448 can lead to a lysogenic conversion of host strain LT4. PMID- 16347411 TI - Rhizobacteria of maize and their antifungal activities. AB - During the growing season of 1984, the rhizobacteria (including organisms from the rhizosphere soil, the rhizoplane, and internal root zones) of 47 maize plants (two varieties) sampled from different locations in France and at different growth stages were inventoried. Isolates were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of their total cell proteins and were found to represent 352 different protein electrotypes. Maize seedlings were initially colonized by a small number of different strains. Densities reached up to 10 CFU/g of root. Later in the season, the population density decreased but the heterogeneity of the rhizobacterial populations increased. Fluorescent pseudomonads represented up to 35% of the total rhizobacterial population and comprised 43 different electrotypes. Other bacteria regularly present were Xanthomonas maltophilia, Serratia liquefaciens, Pseudomonas paucimobilis, and Bacillus spp. There was a very low similarity between rhizobacterial populations of plants of the same cultivar (LG5) within one field at different growth stages and also between rhizobacterial populations of the cultivars LG5 and BRIO42 on the same field. Most electrotypes (76%) were found on a single occasion. None of the 352 electrotypes was present on all plants. In the 1985 analysis the rhizobacteria of maize seedlings (one variety) sampled from one field were characterized. They represented 236 different protein electrotypes. Thirty-three isolates showed antifungal activity against major maize pathogens; they comprised four Pseudomonas cepacia strains, producing pyrrolnitrin as well as another unknown antifungal compound. PMID- 16347412 TI - Ecology of Indigenous Soil Rhizobia: Response of Bradyrhizobium japonicum to Readily Available Substrates. AB - Populations of indigenous Bradyrhizobium japonicum serocluster 123 and serogroups 110 and 138 were studied after various sugars were added to their soil habitat. Loam soil with approximately 10 cells of each group per g of soil were amended every 3 days with 0.1% glucose, sucrose, arabinose, xylose, or galactose. Enumerations of the populations were made every 12 days by immunofluorescence assay. Each B. japonicum population in the sugar-treated soils increased by about 1 log during the first 12 days, to a maximum of about 10 cells by day 36 or 48, irrespective of the sugar added. Maximum growth rates were similar for each group and occurred during the 12-day incubation period. The most rapid growth was in response to arabinose, with a mean generation time of about 3.0 days. Other mean doubling times were 4.0 days with glucose and galactose treatments, 4.5 days with xylose treatment, and 5.4 days with sucrose amendment. These data provide the first direct evidence that indigenous soil rhizobia can compete successfully with other soil bacteria for readily available substrates in soil in the absence of host legume roots or other rhizospheres. The growth rates in soil of the specific B. japonicum populations studied were nearly the same with a given sugar treatment but varied considerably with different sugars. The mean generation times of 3 to 5 days are among the first reported growth rates for heterotrophic bacteria in natural soil. PMID- 16347413 TI - High-pressure equipment for growing methanogenic microorganisms on gaseous substrates at high temperature. AB - High-pressure, high-temperature investigations on thermophilic microorganisms that grow on hydrogen or other gaseous substrates require instrumentation which provides sufficient substrate for cell proliferation up to 2 x 10 to 3 x 10 cells per ml under isothermal and isobaric conditions. To minimize H(2) leakage and to optimize reproducibility at high pressure and high temperature, 10-ml nickel tubes with a liquid/gas ratio of 1:2 were used in a set of autoclaves connected in series. By applying a hydraulic pump and a 2.5-kW heating device, fast changes in temperature (up to 400 degrees C) and pressure (up to 400 MPa) can be accomplished within less than 10 min. To quantify bacterial growth, determinations of cell numbers per unit volume yielded optimum accuracy. Preliminary experiments with the thermophilic, methanogenic archaebacterium Methanococcus thermolithotrophicus showed that bacterial growth depends on both temperature and pressure. At the optimum temperature, increased hydrostatic pressure up to 50 MPa enhanced the growth yield; at a pressure of >75 MPa, cell lysis dominated. Changes in cell proliferation were accompanied by changes in morphology. PMID- 16347414 TI - Properties of Alanine Dehydrogenase and Aspartase from Propionibacterium freudenreichii subsp. shermanii. AB - During lactate fermentation by Propionibacterium freudenreichii subsp. shermanii ATCC 9614, the only amino acid metabolized was aspartate. After lactate exhaustion, alanine was one of the two amino acids to be metabolized. For every 3 mol of alanine metabolized, 2 mol of propionate, 1 mol each of acetate and CO(2), and 3 mol of ammonia were formed. The specific activity of alanine dehydrogenase was 0.08 U/mg of protein during lactate fermentation, and it increased to 0.9 U/mg of protein after lactate exhaustion. Alanine dehydrogenase and aspartase, key enzymes in the metabolism of alanine and aspartate, respectively, were partially purified, and some of their properties were studied. Alanine dehydrogenase had a pH optimum of 9.2 to 9.6 and high K(m) values for both NAD (1 to 4 mM) and alanine (7 to 20 mM). Activity was inhibited by low concentrations of pyruvate and NADH. The pH optimum of aspartase decreased from approximately 7.5 to approximately 6.4 when the MgCl(2) and aspartate concentrations were decreased. Plots of aspartate concentration versus activity showed either hyperbolic or sigmoidal kinetics (interaction coefficient, up to a value of 3.1), depending on pH and MgCl(2) concentration. MgCl(2) was either an activator or an inhibitor, depending on pH and its concentration. Aspartase activity was inhibited by low concentrations of fumarate. The properties of alanine dehydrogenase and aspartase are consistent with the finding that aspartate is metabolized during lactate fermentation, while alanine is only fermented after lactate exhaustion and then at a slow rate. PMID- 16347415 TI - Growth of Thiobacillus ferrooxidans on Elemental Sulfur. AB - Growth kinetics of Thiobacillus ferrooxidans in batch cultures, containing prills of elementary sulfur as the sole energy source, were studied by measuring the incorporation of radioactive phosphorus in free and adsorbed bacteria. The data obtained indicate an initial exponential growth of the attached bacteria until saturation of the susceptible surface was reached, followed by a linear release of free bacteria due to successive replication of a constant number of adsorbed bacteria. These adsorbed bacteria could continue replication provided the colonized prills were transferred to fresh medium each time the stationary phase was reached. The bacteria released from the prills were unable to multiply, and in the medium employed they lost viability with a half-life of 3.5 days. The spreading of the progeny on the surface was followed by staining the bacteria on the prills with crystal violet; this spreading was not uniform but seemed to proceed through distortions present in the surface. The specific growth rate of T. ferrooxidans ATCC 19859 was about 0.5 day, both before and after saturation of the sulfur surface. The growth of adsorbed and free bacteria in medium containing both ferrous iron and elementary sulfur indicated that T. ferrooxidans can simultaneously utilize both energy sources. PMID- 16347416 TI - Chemically Defined Medium for the Accumulation of Intracellular Malate Dehydrogenase by Streptomyces aureofaciens. AB - A chemically defined medium was developed for the production of intracellular malate dehydrogenases by Streptomyces aureofaciens NRRL-B 1286. The composition of the medium (per liter) was as follows: 50 g of starch, 4 g of ammonium sulfate, 7.32 g of l-aspartic acid, 13.8 g of MgSO(4) . 7H(2)O, 1.7 g of K(2)HPO(4), 0.01 g of ZnSO(4) . 7H(2)O, 0.01 g of FeSO(4) . 7H(2)O, 0.01 g of MnSO(4) . H(2)O, and 0.005 g of CoSO(4) . 7H(2)O. The pH of the medium was adjusted to 6.7 to 7.0 after sterilization. The activity of the intracellular malate dehydrogenases of the crude cell extract was greatest after 40 h of mycelium growth in a rotary shaker at 30 degrees C. The best temperature for the enzyme reactions was approximately 35 degrees C for NAD activity at pH 9.7 and 40 degrees C for NADP -linked enzyme at pH 9.0. The NAD activity required Mg, and both activities were sensitive to SH-group reagents. The NADP -dependent activity remained completely stable, and the NAD -dependent activity decreased to a very low residual level after 30 min at 60 degrees C. PMID- 16347417 TI - Role of Chemotaxis in Solvent Production by Clostridium acetobutylicum. AB - The motility of Clostridium acetobutylicum has been investigated during a typical batch fermentation process for solvent production. The motility is characterized by "runs" during the early phase of sugar utilization and acid production, but this changes to "tumbles" during the onset of solventogenesis. Sugars and undissociated acetic and butyric acids have been shown to be attractants for the bacterium, while acetone, butanol, ethanol, and dissociated acetate and butyrate are repellents. It is suggested that chemotactic responses explain why highly motile cells are strongly solventogenic. PMID- 16347418 TI - Hyphal Elongation of Glomus fasciculatus in Response to Root Exudates. AB - The spore germination rates on water agar of the vesicular-arbuscular mycorrhizal fungus Glomus fasciculatus were highest at water potentials of -4 to -6 bars. Root exudates from plants grown in a sterile nutrient solution, with or without phosphorus, did not affect germination. Root exudates collected from 2-, 4-, and 6-week-old Trifolium repens cv. ;Ladino' seedlings that were deprived of P enabled hyphal growth from germinated Glomus fasciculatus spores of 21.4, 14.7, and 7.6 mm, respectively. Hyphal elongation in the presence of exudates from plants grown with P, or in the absence of exudates, was negligible (<1 mm). Root P at 2 weeks was not significantly different between plants grown with and without P. There were no significant differences between the quantities of exudates from plants grown with or without P at 2, 4, and 6 weeks. The data suggest that it is the quality of exudates from plants experiencing P deprivation that is important in stimulating vesicular-arbuscular mycorrhizal hyphal elongation. PMID- 16347419 TI - Relationship between Sodium Influx and Salt Tolerance of Nitrogen-Fixing Cyanobacteria. AB - The relationship between sodium uptake and cyanobacterial salt (NaCl) tolerance has been examined in two filamentous, heterocystous, nitrogen-fixing species of Anabaena. During diazotrophic growth at neutral pH of the growth medium, Anabaena sp. strain L-31, a freshwater strain, showed threefold higher uptake of Na than Anabaena torulosa, a brackish-water strain, and was considerably less salt tolerant (50% lethal dose of NaCl, 55 mM) than the latter (50% lethal dose of NaCl, 170 mM). Alkaline pH or excess K (>25 mM) in the medium causes membrane depolarization and inhibits Na influx in both cyanobacteria (S. K. Apte and J. Thomas, Eur. J. Biochem. 154:395-401, 1986). The presence of nitrate or ammonium in the medium caused inhibition of Na influx accompanied by membrane depolarization. These experimental manipulations affecting Na uptake demonstrated a good negative correlation between Na influx and salt tolerance. All treatments which inhibited Na influx (such as alkaline pH, K above 25 mM, NO(3), and NH(4)), enhanced salt tolerance of not only the brackish-water but also the freshwater cyanobacterium. The results indicate that curtailment of Na influx, whether inherent or effected by certain environmental factors (e.g., combined nitrogen, alkaline pH), is a major mechanism of salt tolerance in cyanobacteria. PMID- 16347420 TI - Fractionation of Stable Carbon Isotopes during Chemoautotrophic Growth of Sulfur Oxidizing Bacteria. AB - Laboratory-grown strains of chemoautotrophic Thiomicrospira sp. strain L-12 and Thiobacillus neapolitanus produced cell carbon that was 24.6 to 25.1 ppt (24.6 to 25.1 mg/g) lower in C isotope abundance than the ambient source of carbon dioxide and bicarbonate. This degree of C isotope depletion was comparable to that found in organic material produced in deep-sea hydrothermal-vent communities. PMID- 16347421 TI - Transposon Mutants of Bradyrhizobium japonicum Altered in Attachment to Host Roots. AB - Transposon mutants of Bradyrhizobium japonicum 110 ARS were produced and screened for changes in attachment ability. Mutant CFK4 produced twice as many piliated cells, attached in 2.5-fold-higher numbers to soybean root segments, and colonized roots in about 2-fold-higher numbers than did the parental strain, 110 ARS. Mutants CFK35 and CFK38 were reduced in their attachment about 2-fold and 3.5-fold, respectively. This corresponded to reductions in piliated cells in their populations, reduced reaction with anti-pilus antiserum, and reduced hydrophobic attachment. Mutants CFK4 and CFK38 nodulated soybeans at about the same level as the parent strain, but CFK35 induced only pseudonodules. Two dimensional gel analyses of the proteins from the mutants showed relatively few changes in proteins. PMID- 16347422 TI - Use of chloroflexus-specific antiserum to evaluate filamentous bacteria of a hot spring microbial mat. AB - Polyclonal antiserum prepared against Chloroflexus aurantiacus reacted with all Chloroflexus strains examined but not with other morphologically or physiologically similar bacteria. Only one of three filament types in a natural hot spring cyanobacterial mat reacted with this antiserum. Reacting filaments remained antigenically positive deep within the mat in material estimated to be several years old. PMID- 16347423 TI - Inadequacy of the eucaryote inhibitor cycloheximide in studies of protozoan grazing on bacteria at the freshwater-sediment interface. AB - Four guilds from a lake sediment-water interface microbial community were isolated and tested for sensitivity to cycloheximide (0.1 to 200 mg liter). Field experiments were conducted to compare the inhibition, dilution, and filtration methods for determining grazing rates. Cycloheximide inhibited anaerobic bacteria at 50 mg liter, and inhibition of bacterial growth was observed in the grazing experiments. The results show that the assumption of selective inhibition of heterotrophic eucaryotes was violated and preclude the use of cycloheximide in grazing experiments. PMID- 16347424 TI - Determining [H]Thymidine Incorporation into Bacterioplankton DNA: Improvement of the Method by DNase Treatment. AB - Determination of [H] thymidine incorporation into bacterial DNA versus other macromolecules is usually achieved by NaOH and hot trichloroacetic acid hydrolysis. This procedure was found not to be specific enough. An alternative method founded on DNase treatment is proposed. Under the new method, the fraction of thymidine incorporated into DNA ranged from 10 to 83%. PMID- 16347425 TI - Oxidation of dimethyl sulfide to dimethyl sulfoxide by phototrophic purple bacteria. AB - Enrichment cultures of phototrophic purple bacteria rapidly oxidized up to 10 mM dimethyl sulfide (DMS) to dimethyl sulfoxide (DMSO). DMSO was qualitatively identified by proton nuclear magnetic resonance. By using a biological assay, DMSO was always quantitatively recovered from the culture media. DMS oxidation was not detected in cultures incubated in the dark, and it was slow in cultures exposed to full daylight. Under optimal conditions, the second-order rate constant for DMS oxidation was 6 day mg of protein ml. The rate constant was reduced in the presence of high concentration of sulfide (>1 mM), but was not affected by the addition of acetate. DMS was also oxidized to DMSO by a pure strain (tentatively identified as a Thiocystis sp.) isolated from the enrichment cultures. DMS supported growth of the enrichment cultures and of the pure strain by serving as an electron source for photosynthesis. A determination of the amount of protein produced in the cultures and an estimation of the electron balance suggested that the two electrons liberated during the oxidation of DMS to DMSO were quantitatively used to reduce carbon dioxide to biomass. The oxidation of DMS by phototrophic purple bacteria may be an important source of DMSO detected in anaerobic ponds and marshes. PMID- 16347426 TI - Fermentation of d-Xylose and l-Arabinose to Ethanol by Erwinia chrysanthemi. AB - Erwinia spp. are gram-negative facultative anaerobes within the family Enterobacteriacae which possess several desirable traits for the conversion of pentose sugars to ethanol, such as the ability to ferment a broad range of carbohydrates and the ease with which they can be genetically modified. Twenty eight strains of Erwinia carotovora and E. chrysanthemi were screened for the ability to ferment d-xylose to ethanol. E. chrysanthemi B374 was chosen for further study on the basis of its superior (4%) ethanol tolerance. We have characterized the fermentation of d-xylose and l-arabinose by the wild type and mutants which bear plasmids containing the pyruvate decarboxylase gene from Zymomonas mobilis. Expression of the gene markedly increased the yields of ethanol (from 0.7 up to 1.45 mol/mol of xylose) and decreased the yields of formate, acetate, and lactate. However, the cells with pyruvate decarboxylase grew only one-fourth as fast as the wild type and tolerated only 2% ethanol. Alcohol tolerance was stimulated by the addition of yeast extract to the growth medium. Xylose catabolism was characterized by a high saturation constant K(s) (4.5 mM). PMID- 16347427 TI - Fermentation of d-Xylose to Ethanol by Genetically Modified Klebsiella planticola. AB - d-Xylose is a plentiful pentose sugar derived from agricultural or forest residues. Enteric bacteria such as Klebsiella spp. ferment d-xylose to form mixed acids and butanediol in addition to ethanol. Thus the ethanol yield is normally low. Zymomonas spp. and most yeasts are unable to ferment xylose, but they do ferment hexose sugars to ethanol in high yield because they contain pyruvate decarboxylase (EC 4.1.1.1), a key enzyme that is absent from enteric bacteria. This report describes the fermentation of d-xylose by Klebsiella planticola ATCC 33531 bearing multicopy plasmids containing the pdc gene inserted from Zymomonas mobilis. Expression of the gene markedly increased the yield of ethanol to 1.3 mol/mol of xylose, or 25.1 g/liter. Concurrently, there were significant decreases in the yields of formate, acetate, lactate, and butanediol. Transconjugant Klebsiella spp. grew almost as fast as the wild type and tolerated up to 4% ethanol. The plasmid was retained by the cells during at least one batch culture, even in the absence of selective pressure by antibiotics to maintain the plasmid. Ethanol production was 31.6 g/liter from 79.6 g of mixed substrate per liter chosen to simulate hydrolyzed hemicellulose. The physiology of the wild type of K. planticola is described in more detail than in the original report of its isolation. PMID- 16347428 TI - Evaluation of Pseudomonas fluorescens for Suppression of Sheath Rot Disease and for Enhancement of Grain Yields in Rice (Oryza sativa L.). AB - Pseudomonas fluorescens strains antagonistic to Sarocladium oryzae, the sheath rot (Sh-R) pathogen of rice (Oryza sativa L.), were evaluated in greenhouse and field tests for suppression of Sh-R severity and enhancement of grain yields of rice. Imprints of rice seedlings and a direct-observation technique of staining roots with fluorochromes confirmed the association of P. fluorescens with roots and the ability of the strain to move along shoot tips. In greenhouse tests, P. fluorescens-treated rice plants (cv. IR 20) showed a 54% reduction in the length of Sh-R lesions. In three field tests, treatment with P. fluorescens reduced the severity of Sh-R by 20 to 42% in five rice cultivars. Bacterization of rice cultivars with P. fluorescens enhanced plant height, number of tillers, and grain yields from 3 to 160%. PMID- 16347429 TI - Increased Effectiveness of Competitive Rhizobium Strains upon Inoculation of Cajanus cajan. AB - A field study was conducted in lysimeters containing N-enriched soil to determine the effects of four competitive rhizobium strains upon yield parameters of pigeon peas (Cajanus cajan). The greatest differences observed were in seed yields; strain P132 effected the highest seed yield (121 +/- 20 g per plant), and the control strain (indigenous rhizobia) effected the lowest yield (43.9 +/- 8 g per plant). With the exception of seeds and pods, the dry matter weights were not different. Although there appeared to be no effect by inoculum strains on the fractional content of N derived from biological nitrogen fixation when the total plant biomass was considered, strains P132 and 401 partitioned more of the N derived from fixation into seeds and leaves than did the other strains. Because the seeds comprised the major portion of plant N, more total N and more N derived from biological nitrogen fixation (about half of total N) were found in plants inoculated with P132, whereas the smallest amount was found in the uninoculated controls. P132 was also the best competitor with respect to indigenous rhizobia and acounted for all of the nodules found on the plants in which it was inoculated. PMID- 16347430 TI - Algal and Bacterial Activities in Acidic (pH 3) Strip Mine Lakes. AB - Reservoir 29 and Lake B are extremely acid lakes (epilimnion pHs of 2.7 and 3.2, respectively), because they receive acidic discharges from coal refuse piles. They differ in that the pH of profundal sediments in Reservoir 29 increased from 2.7 to 3.8 during the period of thermal stratification, whereas permanently anoxic sediments in Lake B had a pH of 6.2. The pH rise in Reservoir 29 sediments was correlated with a temporal increase in H(2)S concentration in the anaerobic hypolimnion from 0 to >1 mM. The chlorophyll a levels in the epilimnion of Reservoir 29 were low, and the rate of primary production was typical of an oligotrophic system. However, there was a dense 10-cm layer of algal biomass at the bottom of the metalimnion. Production by this layer was low owing to light limitation and possibly H(2)S toxicity. The specific photosynthetic rates of epilimnetic algae were low, which suggests that nutrient availability is more important than pH in limiting production. The highest photosynthetic rates were obtained in water samples incubated at pH 2.7 to 4. Heterotrophic bacterial activity (measured by [C]glucose metabolism) was greatest at the sediment/water interface. Bacterial production (assayed by thymidine incorporation) was as high in Reservoir 29 as in a nonacid mesotrophic Indiana lake. PMID- 16347431 TI - Influence of Lime and Phosphate on Nodulation of Soil-Grown Trifolium subterraneum L. by Indigenous Rhizobium trifolii. AB - Previous research had identified four serogroups of Rhizobium trifolii indigenous to the acidic Abiqua soil (fine, mixed, mesic Cumulic Ultic Haploxeroll). Nodulation of subterranean clover (Trifolium subterraneum L.) by two of the serogroups, 6 and 36, was differentially influenced by an application of CaCO(3) which raised the pH of the soil from 5.0 to 6.5. These studies were designed to characterize this phenomenon more comprehensively. Liming the soil with either CaCO(3), Ca(OH)(2), MgO, or K(2)CO(3) significantly (P = 0.05) increased the percent nodule occupancy by serogroup 36, whereas the percent nodule occupancy by serogroup 6 was decreased, but the decrease was significant (P = 0.05) only after application of either CaCO(3) or Ca(OH)(2). Application of KH(2)PO(4) (25 mg of P kg of soil), which did not change soil pH, also significantly (P = 0.05) increased the percent nodule occupancy by serogroup 36. Application of KH(2)PO(4) in combination with Ca(OH)(2) produced the same increase in nodule occupancy by serogroup 36 as did individual application of the two materials. Soil populations of serogroup 36 consistently, and in the majority of cases significantly (P = 0.05), outnumbered those of serogroup 6 before planting and after harvest regardless of soil treatment or the outcome of nodulation. Soil chemical and plant analyses provided no evidence that liming was simulating phosphate addition by increasing the availability and subsequent uptake of soil P(i) by the subclover plants. Liming did, however, result in a significant transformation (30 to 50 mg of P kg of soil) of P(i) from the residual soil P(i) fraction into an NaOH-extractable organic P fraction during the preplant equilibration period. PMID- 16347432 TI - Influence of Phosphate on the Growth and Nodulation Characteristics of Rhizobium trifolii. AB - The growth and nodulating characteristics of Rhizobium trifolii 6 and 36 differed under different external phosphate conditions. Under growth conditions designed to deplete the internal phosphate content of the rhizobia, strain 6 maintained a generation time of 5 h during the exponential phase over two cycles of growth in phosphate-depleted medium. In contrast, the generation time of strain 36 was extended from 3.5 to 9.8 h over two cycles of phosphate-depleted growth, although the organism eventually achieved the same cell density and cellular phosphate content as that of strain 6 at stationary phase. Phosphate-depleted strain 6 required 0.51 +/- 0.08 muM phosphate to commence proliferation, whereas phosphate depleted strain 36 required 0.89 +/- 0.04 muM phosphate under the same conditions. Phosphate-depleted strain 6 maintained viability when exposed to external phosphate concentrations subcritical for growth to occur, whereas phosphate-depleted strain 36 lost viability within 48 h when exposed to medium containing phosphate at concentrations subcritical for growth. Phosphate-depleted strain 36 was inferior to phosphate-depleted strain 6 at nodulating subterranean clover (Trifolium subterraneum L. cv. Mt. Barker) by taking 2 to 4 days longer to develop nodules in phosphatedepleted plant grown medium at pH 5.5. Nodulation by phosphate-depleted strain 36 was accelerated either by including phosphate in the plant growth medium at pH 5.5 or by raising the solution pH of phosphate-depleted plant growth medium to pH 6.5. External phosphate and pH effects were not observed on the nodulating capabilities of phosphate-depleted strain 6 or on luxury phosphate-grown cells of either strain. Phosphatedepleted strains 6 and 36 proliferated to a similar extent on the rhizoplanes even under stringently low external P(i) concentrations. The phosphatase activities of both phosphate depleted strains were significantly (P = 0.05) higher at pH 6.5 than at pH 5.5, and the activity of strain 6 was significantly higher (P = 0.05) than that of strain 36 at pH 5.5 and 5.0. PMID- 16347433 TI - Microcycle Conidiation and Spore-Carrying Capacity of Colletotrichum gloeosporioides on Solid Media. AB - The effect of spore inoculum density, medium concentration, and temperature on slime-spot formation, spore yield, and mycelium production by Colletotrichum gloeosporioides on agar media were studied with a simple microplate assay. A steady-state spore yield (spore-carrying capacity) independent of inoculum density was reached only on media that supported good fungal growth and sporulation. The spore-carrying capacity was reached earlier, the denser the inoculum. On standard mycological media a high inoculum density (2.5 x 10 spores per ml) resulted in a slimy mass of conidia forming a slime spot, a phenomenon associated with greatly reduced mycelium formation and indicative of microcycle conidiation. In contrast, for a similar inoculum density, enhanced mycelial growth preceded sporulation and overrode slime-spot formation on highly concentrated media; a very low medium concentration resulted in much less mycelium, but spore production was also decreased. Exposure to suboptimal growth temperatures of 36 to 48 degrees C for up to 8 days did not induce microcycle conidiation from inocula that did not form a slime spot at 28 degrees C. PMID- 16347434 TI - Production of bacterial inoculants by direct fermentation on nutrient supplemented vermiculite. AB - When supplemented with a nutrient source and moisture, sterile finely ground vermiculite can be used to directly ferment bacterial cultures to prepare bacterial inoculants. All tested bacterial species, including Rhizobium japonicum, R. phaseoli, R. meliloti, R. leguminosarum, Bacillus megaterium, and several Pseudomonas strains, grew at least 10,000-fold in 1 week at room temperature. The final product was stable and had no special storage or handling requirements. Due to the unique properties of vermiculite, direct fermentation of bacteria on nutrient-supplemented vermiculite offers a reliable process for manufacturing bacterial inoculants. PMID- 16347435 TI - Diel Vertical Movements of the Cyanobacterium Oscillatoria terebriformis in a Sulfide-Rich Hot Spring Microbial Mat. AB - Oscillatoria terebriformis, a thermophilic cyanobacterium, carried out a diel vertical movement pattern in Hunter's Hot Springs, Oreg. Throughout most daylight hours, populations of O. terebriformis covered the surface of microbial mats in the hot spring outflows below an upper temperature limit of 54 degrees C. Upon darkness trichomes moved downward by gliding motility into the substrate to a depth of 0.5 to 1.0 mm, where the population remained until dawn. At dawn the population rapidly returned to the top of the mats. Field studies with microelectrodes showed that the dense population of O. terebriformis moved each night across an oxygen-sulfide interface, entering a microenvironment which was anaerobic and reducing, a dramatic contrast to the daytime environment at the mat surface where oxygenic photosynthesis resulted in supersaturated O(2). Laboratory experiments on motility with the use of sulfide gradients produced in agar revealed a negative response to sulfide at concentrations similar to those found in the natural mats. The motility response may help explain the presence of O. terebriformis below the mat surface at night. The movement back to the surface at dawn appears to be due to a combination of phototaxis, photokinesis, and the onset of oxygenic photosynthesis which consumes sulfide. PMID- 16347436 TI - Enhanced Survival of the Cyanobacterium Oscillatoria terebriformis in Darkness under Anaerobic Conditions. AB - Oscillatoria terebriformis, a thermophilic cyanobacterium, maintained viability in darkness under anaerobic conditions by fermenting exogenous glucose or fructose to lactic acid. The time period of survival was greatly extended when the environmental redox potential was lowered by the addition of sodium thioglycolate or titanium(III) citrate. When exposed to aerobic conditions in darkness, many trichomes underwent lysis in 6 h, and death of all cells occurred in 2 to 3 days. The endogenous aerobic respiration rate was high, and the limited dark aerobic survival period appeared to be due to depletion of stored glycogen. Fructose or glucose did not support or increase aerobic respiration in darkness or lengthen aerobic survival time. Enhanced survival of O. terebriformis in darkness under anaerobic, reducing conditions correlates well with the natural nighttime position of this species within sulfide-rich microbial mats associated with hot springs of western North America. PMID- 16347437 TI - Na-Stimulated Transport of l-Methionine in Brevibacterium linens CNRZ 918. AB - The transport of l-methionine by the gram-positive species Brevibacterium linens CNRZ 918 is described. The one transport system (K(m) = 55 muM) found is constitutive for l-methionine, stereospecific, and pH and temperature dependent. Entry of l-methionine into cells is controlled by the internal methionine pool. Competition studies indicate that l-methionine and alpha-aminobutyric acid share a common carrier for their transport. Neither methionine derivatives substituted on the amino or carboxyl groups nor d-methionine was an inhibitor, whereas powerful inhibition was shown by l-cysteine, s-methyl-l-cysteine, dl selenomethionine and dl-homocysteine. Sodium plays important and varied roles in l-methionine transport by B. linens CNRZ 918: (i) it stimulates transport without affecting the K(m), (ii) it increases the specific activity (on a biomass basis) of the l-methionine transport system when present with methionine in the medium, suggesting a coinduction mechanism. l-Methionine transport requires an exogenous energy source, which may be succinic, lactic, acetic, or pyruvic acid but not glucose or sucrose. The fact that l-methionine transport was stimulated by potassium arsenate and to a lesser extent by potassium fluoride suggests that high-energy phosphorylated intermediates are not involved in the process. Monensin eliminates stimulation by sodium. Gramicidin and carbonyl cyanide-m chlorophenylhydrazone act in the presence or absence of Na. N-Ethylmaleimide, p chloromercurobenzoate, valinomycin, sodium azide, and potassium cyanide have no or only a partial inhibitory effect. These results tend to indicate that the proton motive force reinforced by the Na gradient is involved in the mechanism of energy coupling of l-methionine transport by B. linens CNRZ 918. Thus, this transport is partially similar to the well-described systems in gram-negative bacteria, except for the role of sodium, which is very effective in B. linens, a species adapted to the high sodium levels of its niche. PMID- 16347438 TI - Occurrence and Growth of Killer Yeasts during Wine Fermentation. AB - Sixteen wine fermentations were examined for the presence of killer yeasts. Killer property and sensitivity to killer action were found in isolates of Saccharomyces cerevisiae but not in isolates of Kloeckera, Candida, Hansenula, and Torulaspora spp. Several killer and killer-sensitive strains of S. cerevisiae were differentiated by colony morphology, and this property was used to monitor their growth kinetics in mixed cultures in grape juice. Killer-sensitive strains died off within 24 to 48 h during mixed-strain fermentation. Killer action was demonstrated at pH 3.0 and pH 3.5 and over the range of 15 to 25 degrees C but depended on the proportion of killer to killer-sensitive cells at the commencement of fermentation. The dominance of killer strains in mixed-strain fermentations was reflected in the production of ethanol, acetic acid, and glycerol. PMID- 16347439 TI - Sporotrichum thermophile Growth, Cellulose Degradation, and Cellulase Activity. AB - The activity of components of the extracellular cellulase system of the thermophilic fungus Sporotrichum thermophile showed appreciable differences between strains; beta-glucosidase (EC 3.2.1.21) was the most variable component. Although its endoglucanase (EC 3.2.1.4) and exoglucanase (EC 3.2.1.91) activities were markedly lower, S. thermophile degraded cellulose faster than Trichoderma reesei. The production of beta-glucosidase lagged behind that of endoglucanase and exoglucanase. The latter activities were produced during active growth. When growth was inhibited by cycloheximide treatment, the hydrolysis of cellulose was lower than in the control in spite of the presence of both endoglucanase and exoglucanase activities in the culture medium. Degradation of cellulose was a growth-associated process, with cellulase preparations hydrolyzing cellulose only to a limited extent. The growth rate and cell density of S. thermophile were similar in media containing cellulose or glucose. A distinctive feature of fungal development in media incorporating cellulose or lactose (inducers of cellulase activity) was the rapid differentiation of reproductive units and autolysis of hyphal cells to liberate propagules which were capable of renewing growth immediately. PMID- 16347441 TI - Identification of a vesicular-arbuscular mycorrhizal fungus by using monoclonal antibodies in an enzyme-linked immunosorbent assay. AB - Spore morphology is currently used to identify species of vesicular-arbuscular mycorrhizal fungi. We report the first use of a highly specific immunological method for identification of a vesicular-arbuscular mycorrhizal fungus. Two monoclonal antibodies were produced against Glomus occultum. Monoclonal antibodies reacted strongly with both spores and hyphae in an indirect enzyme linked immunosorbent assay. All other mycorrhizal (29 species) and nonmycorrhizal (5 species) fungi tested were nonreactive with the monoclonal antibodies. A single spore of G. occultum was detectable in the presence of high numbers of spores of other vesicular-arbuscular mycorrhizal fungi. Variation in the reaction of G. occultum isolates from West Virginia, Florida, and Colombia suggests that monoclonal antibodies may differentiate strains. PMID- 16347440 TI - Toluene induction and uptake kinetics and their inclusion in the specific affinity relationship for describing rates of hydrocarbon metabolism. AB - The kinetics of concentration-dependent toluene metabolism were examined by evaluating each term in the second-order rate equation. Marine and freshwater pseudomonads were used. Uptake for Pseudomonas sp. strain T2 was characterized by a completely saturatable system with small transport constant (K(t) = 44 mug/liter) and large specific affinity. Kinetics for Pseudomonas putida PpF1 were similar. Induction had little effect on K(t), but it caused the specific affinity to increase from about 0.03 to 320 liters/g of cells per h. The level of induction depended on the time of exposure, the concentration of inducer, and the initial level of induction. If loss of the inducible system was not severe, toluene caused a linear increase in specific affinity with time, and the maximal value achieved at intermediate times (1 to 3 days) was hyperbolic with concentration when K(ind) was 96 mug/liter (A. T. Law and D. K. Button, Appl. Environ. Microbiol. 51:469-476, 1986). As repression became complete, specific affinities were greatly reduced. Then induction required higher toluene concentrations and longer times, and the shape of the specific-affinity curve became sigmoidal with concentration. Cell yields (0.10 to 0.17 g of cells per g of toluene used) were low owing to liberation of organic products: 2-hydroxy-6 oxohepta-2,4-dienoic acid, toluene dihydrodiol, 3-methylcatechol, acetate, formate, and possibly pyruvate, which in turn caused lower rates of growth. Michaelis constants for the reaccumulation of products exceeded those for toluene, but specific affinities were lower and maximal velocities were higher, so that recycling was favored in cultures with high toluene concentration. Although these kinetics predict deviation from the linear relationship between uptake rate and biomass, we could detect none. Effects of saturation and induction were incorporated into the basic specific-affinity relationship. The result appears to be an improvement in the equation used for describing the kinetics of uptake and growth. PMID- 16347442 TI - Characterization of Macromolecular Flocculants Produced by Phormidium sp. Strain J-1 and by Anabaenopsis circularis PCC 6720. AB - Several benthic cyanobacteria were found to produce significant amounts of extracellular flocculants. The macromolecular flocculants produced by Phormidium sp. strain J-1 and Anabaenopsis circularis PCC 6720 were characterized. The Phormidium flocculant is a sulfated heteropolysaccharide to which fatty acids and protein are bound. The polysaccharide backbone is composed of uronic acids, rhamnose, mannose, and galactose. The A. circularis flocculant is also an acidic polysaccharide containing keto acid residues and neutral sugars, but to which no fatty acids, proteins, or sulfates are linked. Both flocculants could be recovered from growth medium by precipitation with cetyltrimethylammonium bromide and were found to bind the cationic dye Alcian-blue in a linear proportion to their concentration in solution. The latter property was used to quantify flocculant concentrations in culture supernatants and natural water samples and to compute their anion densities. PMID- 16347443 TI - Purification and Characterization of an Endoglucanase from Pseudomonas solanacearum. AB - Comparative analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of culture supernatants of a virulent Pseudomonas solanacearum strain and of a spontaneous avirulent mutant derived from it was performed. The results show that the levels of two major polypeptides with molecular masses of 43 and 25 kilodaltons (kDa) were markedly reduced in the spent culture medium of the avirulent mutant. In addition, enzyme assays showed that the level of carboxymethyl cellulase (endoglucanase) activity in the culture supernatants of the avirulent mutant was reduced over 25-fold, whereas polygalacturonase activities in both strains were nearly identical. Purification of the endoglucanase from the spent culture medium of the virulent P. solanacearum strain by adsorption to phosphocellulose, salt elution, and gel-filtration chromatography yielded a >95% pure preparation of the 43-kDa polypeptide. The kinetic and enzymatic properties of the purified endoglucanase were subsequently analyzed. Antibody prepared against the purified 43-kDa endoglucanase was used to demonstrate its production by several strains of P. solanacearum races 1 and 2. PMID- 16347444 TI - Influence of Molecular Size and Ligninase Pretreatment on Degradation of Lignins by Xanthomonas sp. Strain 99. AB - The purpose of this study was to examine the relationship between the molecular size of lignin in several preparations and extent of degradation (mineralization) by Xanthomonas sp. strain 99. The influence of ligninase pretreatment was also examined. Five synthetic lignins and one C-methylated spruce lignin were used. The extent of mineralization to CO(2) was greatest for the samples containing the most low-molecular-weight material, and the low-molecular-weight portions were preferentially (or perhaps solely) degraded. Pretreatment of the five synthetic lignins with crude ligninase increased their molecular size and decreased their degradability by the xanthomonad. Pretreatment of the methylated spruce lignin with crude ligninase caused both polymerization and depolymerization but resulted in a net decrease in bacterial degradability. Our results suggest that the xanthomonad can degrade lignins only up to a molecular weight of 600 to 1,000. PMID- 16347445 TI - Mapping of a Bradyrhizobium japonicum DNA Region Carrying Genes for Symbiosis and an Asymmetric Accumulation of Reiterated Sequences. AB - Spontaneous kanamycin-sensitive derivatives were obtained from Bradyrhizobium japonicum (strain 110) carrying Tn5 insertions in symbiotic gene cluster I; the derivatives were shown to have deletions of cluster I plus flanking DNA which was indicated by the absence of different copies of the repeated sequences RSalpha and RSbeta. The deletion endpoints were mapped using cloned wild-type DNA fragments containing RSalpha copies which also served as origins for overlapped cosmid cloning. The majority of the deletions resulted from recombinational fusion of two remote RSalpha copies. Novel types of repeated sequences (RSgamma, RSdelta, and RSepsilon) occurring in 12, 10, and 4 copies per genome were detected. Seven, nine, and three copies of RSgamma, RSdelta, and RSepsilon, respectively, were located near cluster I. It is concluded that the B. japonicum genome has an unusual DNA segment of >230 kilobase pairs characterized by the presence of repeated sequences and genes for symbiotic N(2) fixation. PMID- 16347446 TI - Conservation of a symbiotic DNA region in soybean root nodule bacteria. AB - Bradyrhizobium japonicum USDA 3I1b110 contains a DNA region in which symbiotic genes and many repeated sequences are closely linked. Hybridization analysis revealed that this region was highly conserved in some B. japonicum strains (USDA 24, USDA 122, USDA 123, ATCC 10324, 61A24) but not in others (USDA 76, 61A76, 61A101). The genomic presence of multiple copies of one of the repeated sequences (RSalpha) appeared to be specifically characteristic for soybean root nodule bacteria, including the fast-growing Rhizobium fredii, which carries most of these RSalpha copies on the symbiotic plasmid. PMID- 16347447 TI - C nuclear magnetic resonance studies of propionate catabolism in methanogenic cocultures. AB - Propionate catabolism was monitored in anaerobic cocultures of propionate degrading and methanogenic bacteria. Metabolism was monitored by use of C enriched propionate and succinate. The intermediates identified indicated that the methylmalonyl coenzyme A pathway was used in these cultures. The data also indicated that a transcarboxylation reaction between succinate and propionyl coenzyme A occurred, yielding propionate and methylmalonyl coenzyme A. PMID- 16347448 TI - Betaine Transport Imparts Osmotolerance on a Strain of Lactobacillus acidophilus. AB - Unlike most Lactobacillus acidophilus strains, a specific strain, L. acidophilus IFO 3532, was found to grow in rich medium containing 1 M sodium acetate, KCl, or NaCl. This strain could also grow with up to 1.8 M NaCl or 3 M nonelectrolytes (fructose, xylose, or sorbitol) added. Thus, this strain was tolerant to osmotic pressures up to 2.8 osM. A search for an intracellular solute which conferred osmoprotection led to the identification of glycine betaine (betaine). Betaine was accumulated to high concentrations in cells growing in MRS medium supplemented with 1 M KCl or NaCl. Uptake of [C]betaine by L. acidophilus 3532 cells suspended in buffer was stimulated by increasing the medium osmotic pressure with 1 M KCl or NaCl. The accumulated betaine was not metabolized further; transport was relatively specific for betaine and was dependent on an energy source. Other lactobacilli, more osmosensitive than strain 3532, including L. acidophilus strain E4356, L. bulgaricus 8144, and L. delbrueckii 9649, showed lower betaine transport rates in response to an osmotic challenge than L. acidophilus 3532. Experiments with chloramphenicol-treated L. acidophilus 3532 cells indicated that the transport system was not induced but appeared to be activated by an increase in osmotic pressure. PMID- 16347449 TI - Temperature characteristics of photosynthetic and heterotrophic activities: seasonal variations in temperate microbial plankton. AB - The seasonal variation in temperature characteristics of photosynthetic and heterotrophic activities in the microbial plankton of Bedford Basin, Nova Scotia, was investigated. Measurements were made of the photosynthetic uptake of [C]bicarbonate and its incorporation into cellular protein as well as the heterotrophic uptake of H-labeled amino acids and their incorporation into cellular protein. Activity-temperature curves were analyzed objectively by nonlinear estimation of parameters from various mathematical models. Over the seasonal cycle, the cardinal temperatures and a parameter formally equivalent to the thermodynamic enthalpy of activation for most of the four processes measured were positively correlated with the water temperature. The temperature sensitivity of metabolic activity (i.e., change in activity per unit change in temperature) was indexed by the tangent to the activity-temperature curves. When this index was expressed in dimensionless form by normalization to the scaling factor of the activity-temperature curves, the resulting relative temperature sensitivity, evaluated at the prevailing temperature, proved to be statistically invariant throughout the year. During the height of the spring bloom, the water temperature (-0.3 degrees C) was not so low as to inhibit metabolic activity of either the phytoplankton or the bacterioplankton. The evidence suggests that heterotrophic utilization of products is not suppressed during the spring phytoplankton bloom. PMID- 16347450 TI - Purification and Characterization of a Substrate-Size-Recognizing Metalloendopeptidase from Streptococcus cremoris H61. AB - During the ripening of Gouda-type cheese, two kinds of endopeptidases were found to participate in the degradation of alphas1-CN(f1-23), a specific product from alphas1-casein hydrolyzed by chymosin. One of the endopeptidases, lactic acid bacteria endopeptidase (LEP-II), which can recognize the size of its substrates, has already been purified and characterized (T. R. Yan, N. Azuma, S. Kaminogawa, and K. Yamauchi, Eur. J. Biochem. 163:259-265, 1987). The other endopeptidase, LEP-I, was purified to homogeneity by conventional chromatographic techniques from Streptococcus cremoris H61. The enzyme appeared to be monomeric, with an apparent molecular weight of 98,000, and its isoelectric point was 5.1. For the hydrolysis of alphas1-CN(f1-23), the enzyme had an optimum pH and temperature of 7.0 to 7.5 and 40 degrees C, respectively. Its activity was inhibited by such chelating agents as EDTA and 1,10-phenanthrolin, and it could be fully reactivated by Mn. Inhibitors specific for serine and thiol proteases had no effect on the protease activity. The enzyme showed a high affinity toward the Glu Asn peptide bond of alphas1-CN(f1-23) and alphas1-CN(f91-100) but showed no hydrolysis activity toward alphas1-CN(f1-52), alphas1-CN(61-122), alphas1-CN(136 196), alphas1-casein, beta-casein, kappa-casein, alpha-lactalbumin, and beta lactoglobulin. The K(m) and V(max) of LEP-I for alphas1-CN(f1-23) were 14.2 pM and 139 U, respectively. PMID- 16347451 TI - Seasonal variation in population density and heterotrophic activity of attached and free-living bacteria in coastal waters. AB - The abundance and heterotrophic activity of attached and free-living bacteria were examined seasonally in coastal water. Heterotrophic activity was determined by the uptake of [C]glucose. The density of attached bacteria was always minor, not showing a seasonal variation, whereas the free-living bacteria were more numerous and showed a marked seasonal variation, their density being higher under warmer conditions. The contribution of the attached bacteria to the total assimilation of [C]glucose (from 10 to 38%) was lower than that of the free living bacteria, neither of them showing a seasonal variation. On a cellular basis, attached bacteria were more active, since they assimilated more [C]glucose and showed, under warmer conditions, a higher cellular volume (0.102 versus 0.047 mum). We consider that the factors responsible for these observations were the amount and quality of the particulate material, the different availability of organic matter for the two types of bacteria, and in a fundamental way, the variation in water temperature. PMID- 16347452 TI - Multiplication and Virulence in Plant Tissues of Escherichia coli Clones Producing Pectate Lyase Isozymes PLb and PLe at High Levels and of an Erwinia chrysanthemi Mutant Deficient in PLe. AB - The phytopathogenic enterobacterium Erwinia chrysanthemi strain EC16 produces four isozymes of pectate lyase (PL), an extracellular enzyme that macerates parenchymatous plant tissues and kills plant cells. A 1.8-kilobase EcoRI DNA fragment containing the entire pelE gene was deleted from the E. chrysanthemi chromosome by marker exchange of a cloned fragment that had been modified in vitro. The resulting mutant, UM1001, produced the isozymes PLa, PLb, and PLc, but not PLe. Mutant UM1001 was compared with wild-type E. chrysanthemi, with Escherichia coli JA221, and with JA221 containing expression vectors with cloned pel genes producing high levels of PLe (pPEL748) or PLb (pPEL343) for the ability to multiply and cause symptoms in intact potato tubers. Tubers were injected with less than 100 bacteria per inoculation site and incubated aerobically or anaerobically. While maceration occurred only in anaerobically incubated tubers, all of the bacteria, including nonpectolytic E. coli controls, multiplied substantially under all conditions. E. coli JA221(pPEL748) caused significantly more maceration than E. coli JA221(pPEL343) or wild-type E. chrysanthemi. Mutant UM1001 caused significantly less maceration than the wild-type E. chrysanthemi. The results establish the importance of PLe in the pectolytic arsenal of E. chrysanthemi by demonstrating that production of PLe can enable E. coli to aggressively macerate tuber tissue and that deletion of pelE significantly diminishes the virulence of E. chrysanthemi. PMID- 16347453 TI - Isolation and Characterization of Frankia sp. Strain FaC1 Genes Involved in Nitrogen Fixation. AB - Genomic DNA was isolated from Frankia sp. strain FaC1, an Alnus root nodule endophyte, and used to construct a genomic library in the cosmid vector pHC79. The genomic library was screened by in situ colony hybridization to identify clones of Frankia nitrogenase (nif) genes based on DNA sequence homology to structural nitrogenase genes from Klebsiella pneumoniae. Several Frankia nif clones were isolated, and hybridization with individual structural nitrogenase gene fragments (nifH, nifD, and nifK) from K. pneumoniae revealed that they all contain the nifD and nifK genes, but lack the nifH gene. Restriction endonuclease mapping of the nifD and nifK hybridizing region from one clone revealed that the nifD and nifK genes in Frankia sp. are contiguous, while the nifH gene is absent from a large region of DNA on either side of the nifDK gene cluster. Additional hybridizations with gene fragments derived from K. pneumoniae as probes and containing other genes involved in nitrogen fixation demonstrated that the Frankia nifE and nifN genes, which play a role in the biosynthesis of the iron molybdenum cofactor, are located adjacent to the nifDK gene cluster. PMID- 16347454 TI - Correlation of direct viable counts with heterotrophic activity for marine bacteria. AB - Viable-bacteria counts, heterotrophic activity, and substrate responsiveness of viable bacteria have been used to measure microbial activity. However, the relationship between these parameters is not clear. Thus, the direct viable count (DVC) method was used to analyze seawater samples collected from several different geographical locations. Samples collected from offshore waters of the South China Sea and western Pacific Ocean yielded DVC that indicated the presence of surface and subsurface peaks of viable, substrate-responsive bacteria which could be correlated with turnover rates of amino acids obtained by using uniformly C-labeled amino acids. DVC were always less than total viable counts (acridine orange direct counts), and the DVC subsurface peak occurred close to and within the chlorophyll a zone, suggesting algal-bacterial interactions within the layer. For comparison with the open-ocean samples, selected substrates were used to determine the response of viable bacteria present in seawater samples collected near an ocean outfall of the Barceloneta Regional Waste Treatment Plant, Barceloneta, Puerto Rico. The number of specific substrate-responsive bacteria at the outfall stations varied depending on the substrate used and the sampling location. Changes in the population size or physiological condition of the bacteria were detected and found to be associated with the presence of pharmaceutical waste. PMID- 16347455 TI - Formation and fate of fermentation products in hot spring cyanobacterial mats. AB - The fate of representative fermentation products (acetate, propionate, butyrate, lactate, and ethanol) in hot spring cyanobacterial mats was investigated. The major fate during incubations in the light was photoassimilation by filamentous bacteria resembling Chloroflexus aurantiacus. Some metabolism of all compounds occurred under dark aerobic conditions. Under dark anaerobic conditions, only lactate was oxidized extensively to carbon dioxide. Extended preincubation under dark anaerobic conditions did not enhance anaerobic catabolism of acetate, propionate, or ethanol. Acetogenesis of butyrate was suggested by the hydrogen sensitivity of butyrate conversion to acetate and by the enrichment of butyrate degrading acetogenic bacteria. Accumulation of fermentation products which were not catabolized under dark anaerobic conditions revealed their importance. Acetate and propionate were the major fermentation products which accumulated in samples collected at temperatures ranging from 50 to 70 degrees C. Other organic acids and alcohols accumulated to a much lesser extent. Fermentation occurred mainly in the top 4 mm of the mat. Exposure to light decreased the accumulation of acetate and presumably of other fermentation products. The importance of interspecies hydrogen transfer was investigated by comparing fermentation product accumulation at a 65 degrees C site, with naturally high hydrogen levels, and a 55 degrees C site, where active methanogenesis prevented significant hydrogen accumulation. There was a greater relative accumulation of reduced products, notably ethanol, in the 65 degrees C mat. PMID- 16347456 TI - Diel interactions of oxygenic photosynthesis and n(2) fixation (acetylene reduction) in a marine microbial mat community. AB - Diel variations in N(2) fixation (acetylene reduction), CO(2) fixation, and oxygen concentrations were measured, on three separate occasions, in a marine microbial mat located on Shackleford Banks, North Carolina. Nitrogenase activity (NA) was found to be inversely correlated with CO(2) fixation and, in two of the three diel periods studied, was higher at night than during the day. Oxygen concentrations within the top 3 mm of the mat ranged from 0 to 400 muM on a diel cycle; anaerobic conditions generally persisted below 4 mm. NA in the mat was profoundly affected by naturally occurring oxygen concentrations. Experimentally elevated oxygen concentrations resulted in a significant depression of NA, whereas the addition of the Photosystem II inhibitor 3(3,4-dichlorophenyl)-1,1 dimethylurea decreased oxygen concentrations within the mat and resulted in a significant short-term enhancement of NA. Mat N(2)-fixing microorganisms include cyanobacteria and heterotrophic, photoautotrophic, and chemolithotrophic eubacteria. Measured (whole-mat) NA is probably due to a combination of the NA of each of these groups of organisms. The relative contributions of each group to whole-mat NA probably varied during diel and seasonal (successional) cycles. Reduced compounds derived from photosynthetic CO(2) fixation appeared to be an important source of energy for NA during the day, whereas heterotrophic or chemolithotrophic utilization of reduced compounds appeared to be an important source of energy for NA at night, under reduced ambient oxygen concentrations. Previous estimates of N(2) fixation calculated on the basis of daytime measurements may have seriously underestimated diel and seasonal nitrogen inputs in mat systems. PMID- 16347457 TI - Microbial growth rates and biomass production in a marine sediment: evidence for a very active but mostly nongrowing community. AB - Biomass, nucleic acid synthesis, and specific growth rates of the microbial communities were measured throughout a vertical profile of a coastal marine sediment. The microbial biomass, as determined by ATP concentrations, in the sediment-water interface was over twice that measured in the other horizons of a 10-cm profile. Likewise, biomass carbon production, as determined by DNA synthesis, and the specific growth rate, as determined from the kinetics of [H]ATP pool labeling, were also elevated at the interface. These results indicate that, due to a large and active community in the interface, the greatest amount of microbial activity, growth, and biosynthesis occurs within the first few millimeters of sediment. These results notwithstanding, a combination of two independent techniques established that over 90% of the sediment-water interface community was not actively growing. PMID- 16347458 TI - Role of Amino Acids and Vitamins in Nutrition of Mesophilic Methanococcus spp. AB - In this study we found that autotrophic methanococci similar to Methanococcus maripaludis obtained up to 57% of their cellular carbon from exogenous amino acids. About 85% of the incorporation was into protein. Primarily nonpolar and basic amino acids and glycine were incorporated; only small amounts of acidic and some polar amino acids were taken up. An additional 10% of the incorporation was into the nucleic acid fraction. Because little CO(2) was formed from the C-amino acids, little metabolism of the amino acids occurred. Therefore the growth stimulation by amino acids was probably due to the sparing of anabolic energy requirements. Of the amino acids incorporated, only alanine was also a sole nitrogen source for these methanococci. In contrast, Methanococcus vannielii and "Methanococcus aeolicus" are autotrophic methanococci which did not incorporate amino acids and did not utilize alanine as a sole nitrogen source. Although glutamine served as a sole nitrogen source for the autotrophic methanococci and Methanococcus voltae, a heterotrophic methanococcus, growth was due to chemical deamination in the medium. M. voltae requires leucine and isoleucine for growth. However, these amino acids were not significant nitrogen sources, and alanine was not a sole nitrogen source for the growth of M. voltae. The branched-chain amino acids were not extensively metabolized by M. voltae. Pantoyl lactone and pantoic acid were readily incorporated by M. voltae. The intact vitamin pantothenate was neither stimulatory to growth nor incorporated. In conclusion, although amino acids and vitamins are nutritionally important to both autotrophic and heterotrophic methanococci, generally they are not subject to extensive catabolism. PMID- 16347459 TI - Wood degradation by white rot fungi: cytochemical studies using lignin peroxidase immunoglobulin-gold complexes. AB - Using an anti-lignin peroxidase antiserum-protein A-gold complex, we found lignin peroxidase mainly intracellularly in several white rot fungi colonizing sawdust under laboratory conditions. This enzyme was also present in fungi found in naturally decayed wood. However, in all cases, lignin peroxidase was located mainly inside the fungal cells. Labeled lignin peroxidase did not bind to the lignocellulosic samples tested, with the exception of poplar milled-wood lignin. These results are discussed in relation to the role of lignin peroxidase during wood degradation. PMID- 16347460 TI - Seasonal and spatial variations in mercury methylation and demethylation in an oligotrophic lake. AB - Microbial mercury methylation and methylmercury decomposition were examined in Lake Clara, an oligotrophic northern Wisconsin seepage lake, using radioisotopic tracers. Methylation activity was near background in the water column, was greatest in the profundal surficial sediments, and decreased with depth in sediment cores. Active demethylation occurred in the water column but was variable. Demethylation was greatest in the surficial sediments and decreased slightly with sediment depth. The methylation/demethylation ratio (M/D) was >1 in the water column, exhibited a sharp peak in surface sediments, and decreased in deeper sediments. Methylation and demethylation activity varied in surface sediments collected along a lake transect. The M/D ratio in surface sediments ranged from 1.4 to 5.8. Methylation in attached microbial communities was near background, while demethylation was high. The M/D ratios in the attached communities were all <0.20. Methylation activity in surface sediments incubated at in situ temperature increased from spring to late summer and decreased in the fall. Demethylation increased from early to midsummer and then declined. The M/D ratio in surface sediments increased from mid- to late summer, and decreased in the fall. These results indicate that the greatest potential for methylation in Lake Clara occurs in the surficial sediments and that methylation in surficial sediments is greatest from mid-July through September. In addition, the net rate of methylmercury production may be significantly affected by demethylation. PMID- 16347461 TI - Production and fate of methylated sulfur compounds from methionine and dimethylsulfoniopropionate in anoxic salt marsh sediments. AB - Anoxic salt marsh sediments were amended with dl-methionine and dimethylsulfoniopropionate (DMSP). Microbial metabolism of methionine yielded methane thiol (MSH) as the major volatile organosulfur product, with the formation of lesser amounts of dimethylsulfide (DMS). Biological transformation of DMSP resulted in the rapid release of DMS and only small amounts of MSH. Experiments with microbial inhibitors indicated that production of MSH from methionine was carried out by procaryotic organisms, probably sulfate-reducing bacteria. Methane-producing bacteria did not metabolize methionine. The involvement of specific groups of organisms in DMSP hydrolysis could not be determined with the inhibitors used, because DMSP was hydrolyzed in all samples except those which were autoclaved. Unamended sediment slurries, prepared from Spartina alterniflora sediments, contained significant (1 to 10 muM) concentrations of DMS. Endogenous methylated sulfur compounds and those produced from added methionine and DMSP were consumed by sediment microbes. Both sulfate reducing and methane-producing bacteria were involved in DMS and MSH consumption. Methanogenesis was stimulated by the volatile organosulfur compounds released from methionine and DMSP. However, apparent competition for these compounds exists between methanogens and sulfate reducers. At low (1 muM) concentrations of methionine, the terminal S-methyl group was metabolized almost exclusively to CO(2) and only small amounts of CH(4). At higher (>100 muM) concentrations of methionine, the proportion of the methyl-sulfur group converted to CH(4) increased. The results of this study demonstrate that methionine and DMSP are potential precursors of methylated sulfur compounds in anoxic sediments and that the microbial community is capable of metabolizing volatile methylated sulfur compounds. PMID- 16347462 TI - Ammonium production in sediments inhibited with molybdate: implications for the sources of ammonium in anoxic marine sediments. AB - Ammonium production in the presence of specific inhibitors of sulfate reduction and methanogenesis was investigated in six marine sediments which differed in bulk properties and organic matter input. In all cases, little effect of the inhibitors on ammonium production was observed, although sulfate reduction was suppressed by molybdate. This gives evidence that the processes of fermentation and hydrolysis are of primary importance in ammonium generation at the sites studied. Although sulfate reduction rates may appear to be coupled to ammonium production rates, sulfate reduction does not necessarily contribute directly to generation of ammonium in marine environments. PMID- 16347463 TI - Rapid and pervasive occupation of fallen mangrove leaves by a marine zoosporic fungus. AB - Samples of leaves of red mangrove (Rhizophora mangle) were incubated on an agar medium selective for pythiaceous oomycetes. Leaves on trees above the water did not contain oomycetes. Marine oomycetes, principally Phytophthora vesicula, had colonized leaves within 2 h of leaf submergence, probably finding them by chemotaxis. The frequency of occurrence of P. vesicula in submerged leaves reached 100% within 30 h of submergence. By 43 h most, if not all, parts of leaves were occupied, and surface treatment with a biocide indicated that leaves were occupied internally. Frequencies of P. vesicula remained near 100% through about 2 weeks of submergence and then declined to about 60% in older (>/=4 weeks) leaves. Leaves of white mangrove (Laguncularia racemosa) were also extensively occupied by P. vesicula after falling into the water column, but decaying leaves of turtlegrass (Thalassia testudinum) were not colonized by oomycetes. Ergosterol analysis indicated that the standing crop of living, non-oomycete (ergosterol containing) fungal mass in submerged red-mangrove leaves did not rise above that which had been present in senescent leaves on the tree; decaying turtlegrass leaves had an ergosterol content that was only about 2% of the maximum concentration detected for red-mangrove leaves. These results suggest that oomycetes are the predominant mycelial eucaryotic saprotrophs of mangrove leaves that fall into the water column and that for turtlegrass leaves which live, die, and decompose under submerged conditions, mycelial eucaryotes make no substantial contribution to decomposition. PMID- 16347464 TI - Homology of pCS1 Plasmid Sequences with Chromosomal DNA in Clavibacter michiganense subsp. sepedonicum: Evidence for the Presence of a Repeated Sequence and Plasmid Integration. AB - Restriction fragments of pCS1, a 50.6-kilobase (kb) plasmid present in many strains of Clavibacter michiganense subsp. sepedonicum ("Corynebacterium sepedonicum"), have been cloned in an M13mp11 phage vector. Radiolabeled forms of these cloned fragments have been used as Southern hybridization probes for the presence of plasmid sequences in chromosomal DNA of this organism. These studies have shown that all tested strains lacking the covalently closed circular form of pCS1 contain the plasmid in integrated form. In each case the site of integration exists on a single plasmid restriction fragment with a size of 5.1 kb. Southern hybridizations with these probes have also revealed the existence of a major repeated sequence in C. michiganense subsp. sepedonicum. Hybridizations of chromosomal DNA with deletion subclones of a 2.9-kb plasmid fragment containing the repeated sequence indicate that the size of the repeated sequence is approximately 1.3 kb. One of the copies of the repeated sequence is on the plasmid fragment containing the site of integration. PMID- 16347465 TI - Microbial hydroxylation of 1,4-cineole. AB - Microorganisms were examined for their potential to hydroxylate the oxygenated monoterpene 1,4-cineole. Using gas chromatography and thin-layer chromatography, screening experiments revealed that hydroxylation at position 2 was the most commonly observed microbial transformation reaction. In most microorganisms, the predominant alcohol metabolite was the 2-endo-alcohol isomer. Preparative-scale incubations were conducted in order to isolate and characterize microbial transformation products by comparison of proton nuclear magnetic resonance, mass spectrometry, and chromatography profiles with those of cineole standards. Streptomyces griseus yielded 8-hydroxy-1,4-cineole as the major hydroxylation product together with 2-exo- and 2-endo-hydroxy-1,4-cineoles. PMID- 16347466 TI - A Technique for Predicting the Solvent-Producing Ability of Clostridium acetobutylicum. AB - Changes in colony morphology were associated with the degeneration of solvent producing strains of Clostridium acetobutylicum. The most efficient solvent producing strains gave rise exclusively to colonies with dense centers containing large numbers of spores. Many outgrowths of various morphologies developed from the perimeter of such colonies after several days of incubation. The most degenerate cultures did not produce solvents and gave rise to large diffuse colonies that did not contain spores. These diffuse colonies did not produce outgrowths. Intermediate colony types were also observed. These could be derived from liquid cultures that were relatively poor solvent producers or from the outgrowths of colonies of efficient solvent-producing strains. Some of these intermediate types produced spores but did so less frequently than the high solvent-producing strains. The spores of the intermediate types could not be distinguished from those of the most efficient solvent producers on the basis of heat sensitivity. The relationship observed between colony morphology and solvent production provides a method for predicting the solvent-producing potential of C. acetobutylicum cultures. PMID- 16347467 TI - Spontaneous Disaggregation of Methanosarcina mazei S-6 and Its Use in the Development of Genetic Techniques for Methanosarcina spp. AB - When monomethylamine was the growth substrate, spontaneous disaggregation of Methanosarcina mazei S-6 commenced at the mid-exponential phase and resulted in the formation of a suspension containing 10 to 10 free cells per ml. Free cells were osmotically fragile and amenable to extraction of DNA. Hypertonic media for the manipulation and regeneration of free cells into aggregates were developed, and plating efficiencies of 100% were achieved for M. mazei S-6 and LYC. Free cells of strain S-6 required MgCl(2) (10 mM) for growth, whereas aggregates did not. Specific growth rates of strains S-6 and LYC were increased by MgCl(2). Treatment with pronase caused sphere formation and removal of the protein wall of cells of strain S-6, but protoplasts could not be regenerated. The disaggregating enzyme produced by strain S-6 facilitated the preparation of suspensions of free cells of some strains of Methanosarcina barkeri. Although this provided a means of extracting high-molecular-weight DNA from M. barkeri, less than 0.1% of free cells were viable. PMID- 16347468 TI - Competitive Exclusion of Epiphytic Bacteria by IcePseudomonas syringae Mutants. AB - The growth of ice nucleation-active and near-isogenic ice nucleation-deficient (Ice) Pseudomonas syringae strains coexisting on leaf surfaces was examined to determine whether competition was sufficient to account for antagonism of phylloplane bacteria. The ice nucleation frequency spectra of 46 IceP. syringae mutants, obtained after mutagenesis with ethyl methanesulfonate, differed both quantitatively and qualitatively, but the mutants could be grouped into four distinct phenotypic classes. The numbers of ice nucleation-active bacteria and ice nuclei active at -5 degrees C were reduced on plants colonized with IceP. syringae mutant strains before challenge inoculations with an IceP. syringae wild type strain. Frost injury to plants pretreated with IceP. syringae strains was also reduced significantly compared with that to control plants and was correlated with the population size of the IceP. syringae strain and with the numbers of ice nuclei active at -5 degrees C. An IceP. syringae strain colonized leaves, flowers, and young fruit of pears in field experiments and significantly reduced the colonization of these tissues by IceP. syringae strains and Erwinia amylovora as compared with untreated trees. PMID- 16347469 TI - Flagellar Motility Confers Epiphytic Fitness Advantages upon Pseudomonas syringae. AB - The role of flagellar motility in determining the epiphytic fitness of an ice nucleation-active strain of Pseudomonas syringae was examined. The loss of flagellar motility reduced the epiphytic fitness of a normally motile P. syringae strain as measured by its growth, survival, and competitive ability on bean leaf surfaces. Equal population sizes of motile parental or nonmotile mutant P. syringae strains were maintained on bean plants for at least 5 days following the inoculation of fully expanded primary leaves. However, when bean seedlings were inoculated before the primary leaves had expanded and bacterial populations on these leaves were quantified at full expansion, the population size of the nonmotile derivative strain reached only 0.9% that of either the motile parental or revertant strain. When fully expanded bean primary leaves were coinoculated with equal numbers of motile and nonmotile cells, the population size of a nonmotile derivative strain was one-third of that of the motile parental or revertant strain after 8 days. Motile and nonmotile cells were exposed in vitro and on plants to UV radiation and desiccating conditions. The motile and nonmotile strains exhibited equal resistance to both stresses in vitro. However, the population size of a nonmotile strain on leaves was less than 20% that of a motile revertant strain when sampled immediately after UV irradiation. Epiphytic populations of both motile and nonmotile P. syringae declined under desiccating conditions on plants, and after 8 days, the population size of a nonmotile strain was less than one-third that of the motile parental or revertant strain. PMID- 16347470 TI - Plasmid-Associated Bacteriocin Production and Sucrose Fermentation in Pediococcus acidilactici. AB - Production of bacteriocin activity designated pediocin PA-1 was associated with the presence of a 6.2-megadalton plasmid in Pediococcus acidilactici PAC1.0. The bacteriocin exhibited activity against P. acidilactici, P. pentosaceus, Lactobacillus plantarum, L. casei, L. bifermentans, and Leuconostoc mesenteroides subsp. dextranicum. Partial characterization of pediocin PA-1 is described. The molecular weight of pediocin PA-1 was ca. 16,500. Additionally, strain PAC1.0 was found to contain a 23-megadalton plasmid associated with sucrose-fermenting ability. PMID- 16347471 TI - Conserved Plasmid Hydrogen-Uptake (hup)-Specific Sequences within HupRhizobium leguminosarum Strains. AB - Thirteen Rhizobium leguminosarum strains previously reported as H(2)-uptake hydrogenase positive (Hup) or negative (Hup) were analyzed for the presence and conservation of DNA sequences homologous to cloned Bradyrhizobium japonicum hup specific DNA from cosmid pHU1 (M. A. Cantrell, R. A. Haugland, and H. J. Evans, Proc. Natl. Acad. Sci. USA 80:181-185, 1983). The Hup phenotype of these strains was reexamined by determining hydrogenase activity induced in bacteroids from pea nodules. Five strains, including H(2) oxidation-ATP synthesis-coupled and uncoupled strains, induced significant rates of H(2)-uptake hydrogenase activity and contained DNA sequences homologous to three probe DNA fragments (5.9-kilobase [kb] HindIII, 2.9-kb EcoRI, and 5.0-kb EcoRI) from pHU1. The pattern of genomic DNA HindIII and EcoRI fragments with significant homology to each of the three probes was identical in all five strains regardless of the H(2)-dependent ATP generation trait. The restriction fragments containing the homology totalled about 22 kb of DNA common to the five strains. In all instances the putative hup sequences were located on a plasmid that also contained nif genes. The molecular sizes of the identified hup-sym plasmids ranged between 184 and 212 megadaltons. No common DNA sequences homologous to B. japonicum hup DNA were found in genomic DNA from any of the eight remaining strains showing no significant hydrogenase activity in pea bacteroids. These results suggest that the identified DNA region contains genes essential for hydrogenase activity in R. leguminosarum and that its organization is highly conserved within Hup strains in this symbiotic species. PMID- 16347472 TI - Dynamics of methane production, sulfate reduction, and denitrification in a permanently waterlogged alder swamp. AB - The dynamics of sulfate reduction, methane production, and denitrification were investigated in a permanently waterlogged alder swamp. Molybdate, an inhibitor of sulfate reduction, stimulated methane production in soil slurries, thus suggesting competition for common substrates between sulfate-reducing and methane producing bacteria. Acetate, hydrogen, and methanol were found to stimulate both sulfate reduction and methane production, while trimethylamine mainly stimulated methane production. Nitrate addition reduced both methane production and sulfate reduction, either as a consequence of competition or poisoning of the bacteria. Sulfate-reducing bacteria were only slightly limited by the availability of electron acceptors, while denitrifying bacteria were seriously limited by low nitrate concentrations. Arrhenius plots of the three processes revealed different responses to temperature changes in the slurries. Methane production was most sensitive to temperature changes, followed by denitrification and sulfate reduction. No significant differences between slope patterns were observed when comparing summer and winter measurements, indicating similar populations regarding temperature responses. PMID- 16347473 TI - Effect of Plasmid pSa and of Auxin on Attachment of Agrobacterium tumefaciens to Carrot Cells. AB - When the plasmid pSa is introduced into Agrobacterium tumefaciens, its presence results in the suppression of bacterial virulence. A. tumefaciens(pSa) cells are virulent on Bryophyllum diagremontiana only when inoculated with auxin. A. tumefaciens(pSa) cells also bind to plant cells only in the presence of auxin. The effect of auxin is on the bacteria rather than on the plant cells, since the bacteria require auxin to bind to heat-killed carrot cells. Bacteria containing pSa and grown in the absence of auxin showed a lag in binding to carrot cells in auxin-containing medium. This lag was not seen during the binding of wild-type strains. Tetracycline inhibited the binding of A. tumefaciens(pSa) in auxin containing medium, suggesting that bacterial protein synthesis is required for the auxin effect. No difference was seen in the size or ability to inhibit bacterial binding of lipopolysaccharides from bacteria containing or lacking pSa and grown with or without auxin. A. tumefaciens(pSa) cells grown in the absence of auxin lacked surface polypeptide(s) found in bacteria grown in the presence of auxin and in the wild-type bacteria, which do not contain pSa. Thus, the presence of certain polypeptides appears to be associated with the ability of the bacteria to bind to plant cells. PMID- 16347474 TI - Effect of Proteolytic Enzymes on Transfection and Transformation of Streptococcus lactis Protoplasts. AB - With both chymotrypsin and mutanolysin used to form protoplasts, consistent transformation frequencies of 10 to 10 transformants and transfectants per mug of DNA were achieved. The procedure was used to transform protoplasts of Streptococcus cremoris CS224 at low frequency (5 transformants per mug of DNA). PMID- 16347475 TI - Freeze-Thawing of Aquaspirillum magnetotacticum Cells Selectively Releases Periplasmic Proteins. AB - Cells of the gram-negative bacterium Aquaspirillum magnetotacticum, when suspended in buffer and freeze-thawed, produced pinkish orange supernatant fluid. The fluid contained CO(2) > acetate. The isotopic fractionation for methane derived from acetate was only one-third of that observed with methanol as the substrate. The data presented indicate that the last enzyme of methanogenesis, methylreductase, is not the primary site of isotopic discrimination during methanogenesis from methanol or CO(2). These results also support biogeochemical interpretations that gas produced in environments in which acetate is the primary methane precursor will have higher C/C ratios than those from environments where other substrates predominate. PMID- 16347477 TI - Citrate Cycle Intermediates in the Metabolism of Aspartate and Lactate by Propionibacterium freudenreichii subsp. shermanii. AB - Propionibacterium freudenreichii subsp. shermanii metabolized 7 mol of aspartate to 6 mol of succinate, 4 mol of CO(2), and 7 mol of ammonia. When lactate, sparged with 100% CO(2), was fermented at pH 5.5, unexpectedly high ratios of propionate to acetate were obtained (i.e., 3.2 to 3.8:1). Citrate cycle intermediates may be involved in these fermentations. PMID- 16347478 TI - Role of dissolution rate and solubility in biodegradation of aromatic compounds. AB - [This corrects the article on p. 294 in vol. 53.]. PMID- 16347479 TI - Metabolism of Lignin Model Compounds of the Arylglycerol-beta-Aryl Ether Type by Pseudomonas acidovorans D(3). AB - A natural bacterial isolate that we have classified as Pseudomonas acidovorans grows on the lignin model compounds 1-(3,4-dimethoxyphenyl)-2-(2 methoxyphenoxy)propane-1,3-diol (compound 1) and 1-(4-hydroxy-3-methoxyphenyl)-2 (2-methoxyphenoxy)propane-1,3-diol (compound 1'), as well as on the corresponding 1-oxo compounds (2 and 2') as sole sources of carbon and energy. Metabolic intermediates present in cultures growing on compound 1 included compound 2, 2 methoxyphenol (guaiacol [compound 3]), beta-hydroxypro-pioveratrone (compound 4), acetoveratrone (compound 5), and veratric acid (compound 6). Also identified were compounds 1', 2', beta-hydroxypropiovanillone (compound 4'), and acetovanillone (compound 5'), indicating that 4-O demethylation also occurs. The phenolic intermediates were the same as those found in cultures growing on compound 1'. Compounds 2 and 2' were in part also reduced to compounds 1 and 1', respectively. Compound 3 was shown to be derived from the 2-methoxyphenoxy moiety. A suggested degradation scheme is as follows: compound 1-->2-->(3 + 4)-->5-->6 (and similarly for 1'). In this scheme, the key reaction is cleavage of the ether linkage between C-2 (C(beta)) of the phenylpropane moiety and the 2-methoxyphenoxy moiety in compounds 2 and 2' (i.e., beta-aryl ether cleavage). On the basis of compounds identified, viz., 3 and 4 (4'), cleavage appears formally to be reductive. Because this is unlikely, the initial cleavage products probably were not detected. The implications of these results for the enzyme(s) responsible are discussed. PMID- 16347480 TI - Biogeochemical Conditions Favoring Magnetite Formation during Anaerobic Iron Reduction. AB - Several anaerobic bacteria isolated from the sediments of Contrary Creek, an iron rich environment, produced magnetite when cultured in combinations but not when cultured alone in synthetic iron oxyhydroxide medium. When glucose was added as a carbon source, the pH of the medium decreased (to 5.5) and no magnetite was formed. When the same growth medium without glucose was used, the pH increased (to 8.5) and magnetite was formed. In both cases, Fe was released into the growth medium. Geochemical equilibrium equations with E(h) and pH as master variables were solved for the concentrations of iron and inorganic carbon that were observed in the system. Magnetite was predicted to be the dominant iron oxide formed at high pHs, while free Fe or siderite were the dominant forms of iron expected at low pHs. Thus, magnetite formation occurs because of microbial alteration of the local E(h) and pH conditions, along with concurrent reduction of ferric iron (direct biological reduction or abiological oxidation-reduction reactions). PMID- 16347481 TI - Genetic Diversity in Bradyrhizobium japonicum Serogroup 123 and Its Relation to Genotype-Specific Nodulation of Soybean. AB - The genetic diversity among 20 field isolates of Bradyrhizobium japonicum serogroup 123 was examined by using restriction endonuclease digestions, one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total cell proteins, Southern hybridization analysis of nif and nod genes, and intrinsic antibiotic resistance profiles. All of the isolates were previously separated into three broad nodulation classes (low, medium, and high) based on their ability to form symbioses with specific soybean genotypes. Results of our studies indicate that there is a relationship between these three genotype specific nodulation classes and groupings that have been made based on genomic DNA digestion patterns, sodium dodecyl sulfate-protein profiles, and Southern hybridizations to a nifHD gene probe. Intrinsic antibiotic resistance profiles and nodAB gene hybridizations were not useful in determining interrelationships between isolates and nodulation classes. Southern hybridizations revealed that two of the isolates had reiterated nod genes; however, there was no correlation between the presence of extra nodAB genes and the nodulation classes or symbiotic performance on permissive soybean genotypes. Hybridizations with the nif gene probe indicated that there is a relationship among serogroup, nodulation class, and the physical organization of the genome. PMID- 16347482 TI - Nodulation and Competition for Nodulation of Selected Soybean Genotypes among Bradyrhizobium japonicum Serogroup 123 Isolates. AB - Twenty recently obtained field isolates of Bradyrhizobium japonicum serogroup 123 were tested for their nodule mass production on the standard commercial soybean (Glycine max (L.) Merr. cv. Williams) and on two soybean plant introduction (PI) genotypes previously determined to restrict nodulation by strain USDA 123. Four of the field isolates showed similar restricted nodulation on the two genotypes, while all 20 isolates produced a normal amount of nodules on G. max cv. Williams. Serological analyses with adsorbed fluorescent antibodies showed that members of the 123 serotype ranked low in nodulation of the two PIs, in contrast to members of serotypes 127 and 129. Competition studies on the PIs indicated that isolates which were restricted were not competitive for nodule occupancy against strain USDA 110. However, unrestricted isolates of serogroup 123 were very competitive against USDA 110. On G. max cv. Williams, all serogroup 123 isolates tested were very competitive against USDA 110. PMID- 16347483 TI - Competitive mechanisms for inhibition of sulfate reduction and methane production in the zone of ferric iron reduction in sediments. AB - Mechanisms for inhibition of sulfate reduction and methane production in the zone of Fe(III) reduction in sediments were investigated. Addition of amorphic iron(III) oxyhydroxide to sediments in which sulfate reduction was the predominant terminal electron-accepting process inhibited sulfate reduction 86 to 100%. The decrease in electron flow to sulfate reduction was accompanied by a corresponding increase in electron flow to Fe(III) reduction. In a similar manner, Fe(III) additions also inhibited methane production in sulfate-depleted sediments. The inhibition of sulfate reduction and methane production was the result of substrate limitation, because the sediments retained the potential for sulfate reduction and methane production in the presence of excess hydrogen and acetate. Sediments in which Fe(III) reduction was the predominant terminal electron-accepting process had much lower concentrations of hydrogen and acetate than sediments in which sulfate reduction or methane production was the predominant terminal process. The low concentrations of hydrogen and acetate in the Fe(III)-reducing sediments were the result of metabolism by Fe(III)-reducing organisms of hydrogen and acetate at concentrations lower than sulfate reducers or methanogens could metabolize them. The results indicate that when Fe(III) is in a form that Fe(III)-reducing organisms can readily reduce, Fe(III)-reducing organisms can inhibit sulfate reduction and methane production by outcompeting sulfate reducers and methanogens for electron donors. PMID- 16347484 TI - Initial steps in the pathway for bacterial degradation of two tetrameric lignin model compounds. AB - We investigated the metabolic route by which a lignin tetramer-degrading mixed bacterial culture degraded two tetrameric lignin model compounds containing beta O-4 and 5-5 biphenyl structures. The alpha-hydroxyl groups in the propane chain of both phenolic and nonphenolic tetramers were first oxidized symmetrically in two successive steps to give monoketones and diketones. These ketone metabolites were decomposed through C(alpha)(=O)-C(beta) cleavage, forming trimeric carboxyl acids which were further metabolized through another C(alpha)(=O)-C(beta) cleavage. Dehydrodiveratric acid, which resulted from the cleavage of the carbon bonds of the nonphenol tetramer, was demethylated twice. Four metabolites of the phenolic tetramer were purified and identified. All of these were stable compounds in sterile mineral medium, but were readily degraded by lignin tetramer degrading bacteria along the same pathway as the phenol tetramer. No monoaromatic metabolites accumulated. All metabolites were identified by mass and proton magnetic resonance spectrometry. The metabolic route by which the mixed bacterial culture degraded tetrameric lignin model compounds was different from the route of the main ligninase-catalyzed C(alpha)-C(beta) cleavage by Phanerochaete chrysosporium. PMID- 16347485 TI - Complementary methodologies to identify specific agrobacterium strains. AB - Serological techniques and restriction enzyme cleavage patterns of total DNA were used to differentiate strains of Agrobacterium spp. Forty-five wild-type and plasmid-cured Agrobacterium strains were tested by immunodiffusion and immunofluorescence against polyclonal antisera to a crude ribosome preparation from Agrobacterium strains K84, U11, B6, A323, NT1, and C58. In immunodiffusion gels, these antisera reacted only with water-phenol extracts of the homologous strain, producing a single, strain-specific precipitin line. In contrast, when the same antisera were used in immunofluorescence staining, cross-reactions occurred with a limited number of heterologous Agrobacterium strains. However, the cross-reacting heterologous cells fluoresced generally less brightly than the homologous cells. When the EcoRI-digested DNA profiles from the same Agrobacterium strains were compared, 34 distinct cleavage patterns were observed. The DNA profiles were the same for all strains sharing a common chromosomal background and correlated with the strain-specific serological reaction. The presence or absence of plasmid DNA did not alter the strain-specific serological reaction or the DNA cleavage patterns. Both the serological reaction and the restriction enzyme digestion of total DNA were complementary to each other. These methods were used successfully to identify A. radiobacter K84 strains which were recovered 6 months after being inoculated to young trees in the field. PMID- 16347486 TI - Effects of flow cytometric analysis on morphology and viability of fragile phytoplankton. AB - We assessed damage done to especially delicate marine phytoplankton cells by passage through a Coulter Epics V flow cytometer. The cells did not distort or lyse after exposure to fluidics or to laser light to 1,000 mW. The cells did sustain damage evidenced by temporary growth rate depressions. The four clones tested eventually resumed control growth rates after growth lags to 48 h. PMID- 16347487 TI - Increased Phosphorus Uptake by Wheat and Field Beans Inoculated with a Phosphorus Solubilizing Penicillium bilaji Strain and with Vesicular-Arbuscular Mycorrhizal Fungi. AB - Greenhouse and field experiments were conducted to test the effect of a P solubilizing isolate of Penicillium bilaji on the availability of Idaho rock phosphate (RP) in a calcareous soil. Under controlled greenhouse conditions, inoculation of soils with P. bilaji along with RP at 45 mug of P per g of soil resulted in plant dry matter production and P uptake by wheat (Triticum aestivum) and beans (Phaseolus vulgaris) that were not significantly different from the increases in dry matter production and P uptake caused by the addition of 15 mug of P per g of soil as triple superphosphate. Addition of RP alone had no effect on plant growth. Addition of vesicular-arbuscular mycorrhizal fungi was necessary for maximum effect in the sterilized soil in the greenhouse experiment. Under field conditions, a treatment consisting of RP (20 kg of P per ha of soil) plus P. bilaji plus straw resulted in wheat yields and P uptake equivalent to increases due to the addition of monoammonium phosphate added at an equivalent rate of P. RP added alone had no effect on wheat growth or P uptake. The results indicate that a biological system of RP solubilization can be used to increase the availability of RP added to calcareous soils. PMID- 16347488 TI - Adaptation of aquatic microbial communities to hg stress. AB - The mechanism of adaptation to Hg in four aquatic habitats was studied by correlating microbially mediated Hg volatilization with the adaptive state of the exposed communities. Community diversity, heterotrophic activity, and Hg resistance measurements indicated that adaptation of all four communities was stimulated by preexposure to Hg. In saline water communities, adaptation was associated with rapid volatilization after an initial lag period. This mechanism, however, did not promote adaptation in a freshwater sample, in which Hg was volatilized slowly, regardless of the resistance level of the microbial community. Distribution of the mer operon among representative colonies of the communities was not related to adaptation to Hg. Thus, although volatilization enabled some microbial communities to sustain their functions in Hg-stressed environments, it was not mediated by the genes that serve as a model system in molecular studies of bacterial resistance to mercurials. PMID- 16347489 TI - Annual cycle of bacterial specific biovolumes in howe sound, a canadian west coast fjord sound. AB - The mean specific biovolumes (biovolume cell) of the bacterioplankton within a 250-m-deep water column in Howe Sound, British Columbia, were determined for the period of 4 September 1984 to 23 October 1985. These bacteria had an annual cycle in mean specific biovolume; they were small (ca. 0.058 mum) in mid-winter, larger in spring (ca. 0.076 mum), larger again in summer (up to 0.102 mum), and largest (ca. 0.133 mum) in early fall (immediately after the decrease in phytoplankton production). The mean specific biovolumes changed coincidently through the water column with time, although the larger bacterioplankton tended to occur in the surface and deepest water. Although the mean specific biovolumes correlated better with in situ temperature (r = 0.65, a = 0.01) than with in situ chlorophyll a concentration (r = 0.34, a = 0.25), modeling experiments with batch cultures of the dinoflagellate Prorocentrum minimum (Pavillard) and the green alga Dunaliella tertiolecta (Butcher) indicated that the biomass and physiological condition of the phytoplankters may be more important than temperature in determining these bacterial specific biovolumes. PMID- 16347490 TI - Plant Cell Wall Carbohydrates as Substrates for Azospirillum brasiliense. AB - Carbohydrate components (simple sugars and polysaccharides) of cell walls of pearl millet (Pennisetum americanum L., cv. Gahi) were studied as potential substrates for the root-associated diazotroph Azospirillum brasiliense Sp. 7. Simple sugars were utilized, but no evidence was obtained to support the suggestion that the polysaccharide components tested might serve as substrates for growth following hydrolysis by the associated azospirilla. PMID- 16347491 TI - Preservation of ATP in hypersaline environments. AB - High concentrations of particulate ATP were found in the anoxic brines of the Orca Basin and East Flower Garden, Gulf of Mexico. Other measurements indicative of growth and respiration suggested that the microbial community in the brines was inactive, but somehow the ATP associated with the cells persisted. Conceivably, when cells growing just above the interface sank into the brine, the increased osmotic stress could elicit an osmoregulatory response resulting in increased ATP. It was also possible that hydrolytic enzymes were inactivated, resulting in the preservation of ATP. Experiments in which a culture of marine bacteria was suspended in menstrua of different salinities comparable to those found across the Orca Basin interface revealed that as salinity increased, ATP increased three- to sixfold. Within 24 h the ATP fell to its initial level and remained at that concentration for 3 days, at which time the experiment was terminated. In contrast, the control suspensions, at a salinity of 28% (grams per liter) had 1/10th of the initial ATP concentration when the experiment was ended. Cells were also exposed to killing UV irradiation, enabling us to demonstrate with absolute certainty that cellular ATP could be preserved. At the end of the experiment, the viable component of the population was reduced by orders of magnitude by UV irradiation, but the ATP levels of the cells suspended in brine did not decrease. In certain environments it appears that the conventional analytical tools of the microbial ecologist must be interpreted with caution. PMID- 16347492 TI - Extracellular Enzyme Activities during Lignocellulose Degradation by Streptomyces spp.: A Comparative Study of Wild-Type and Genetically Manipulated Strains. AB - The wild-type ligninolytic actinomycete Streptomyces viridosporus T7A and two genetically manipulated strains with enhanced abilities to produce a water soluble lignin degradation intermediate, an acid-precipitable polymeric lignin (APPL), were grown on lignocellulose in solid-state fermentation cultures. Culture filtrates were periodically collected, analyzed for APPL, and assayed for extracellular lignocellulose-catabolizing enzyme activities. Isoenzymes were analyzed by polyacrylamide gel electrophoresis and activity staining on the gels. Two APPL-overproducing strains, UV irradiation mutant T7A-81 and protoplast fusion recombinant SR-10, had higher and longer persisting peroxidase, esterase, and endoglucanase activities than did the wild-type strain T7A. Results implicated one or more of these enzymes in lignin solubilization. Only mutant T7A 81 had higher xylanase activity than the wild type. The peroxidase was induced by both lignocellulose and APPL. This extracellular enzyme has some similarities to previously described ligninases in fungi. This is the first report of such an enzyme in Streptomyces spp. Four peroxidase isozymes were present, and all catalyzed the oxidation of 3,4-dihydroxyphenylalanine, while one also catalyzed hydrogen peroxide-dependent oxidation of homoprotocatechuic acid and caffeic acid. Three constitutive esterase isozymes were produced which differed in substrate specificity toward alpha-naphthyl acetate and alpha-naphthyl butyrate. Three endoglucanase bands, which also exhibited a low level of xylanase activity, were identified on polyacrylamide gels as was one xylanase-specific band. There were no major differences in the isoenzymes produced by the different strains. The probable role of each enzyme in lignocellulose degradation is discussed. PMID- 16347493 TI - Clostridium acetobutylicum Mutants That Produce Butyraldehyde and Altered Quantities of Solvents. AB - Spontaneous mutants of Clostridium acetobutylicum NRRL B643 that were resistant to allyl alcohol (AA) were selected and characterized. These mutants contained 10 to 100-fold reduced activities of butanol and ethanol alcohol dehydrogenase. The AA mutants formed two groups and produced no ethanol. Type 1 AA mutants produced significant amounts of a new solvent, butyraldehyde, and contained normal levels of the coenzyme A-dependent butyraldehyde dehydrogenase (BAD). Type 2 AA mutants produced no significant butyraldehyde and lower levels of all solvents, and they contained 45- to 100-fold lower activity levels of BAD. Following ethyl methanesulfonate mutagenesis, low-acid-producing (Acid) mutants were selected and characterized as superinduced solvent producers, yielding more than 99% of theoretical glucose carbon as solvents and only small amounts of acetate and butyrate. Following ethyl methanesulfonate mutagenesis, 13 sporulation-negative (Spo) mutants were characterized; and 3 were found to produce only butyrate and acetate, a minor amount of acetone, and no alcohols. These Spo mutants contained reduced butanol dehydrogenase activity and no BAD enzyme activity. The data support the view that the type 2 AA, the Acid, and the Spo mutants somehow alter normal regulated expression of the solvent pathway in C. acetobutylicum. PMID- 16347494 TI - Effects of Temperature and pH on Survival of Free Nuclear Polyhedrosis Virus of Autographa californica. AB - The effects of temperature and low pH on replication and survival of nonoccluded Autographa californica nuclear polyhedrosis virus were investigated. No virus replication or formation of polynuclear inclusion bodies occurred at 37 degrees C. The virus was immediately inactivated upon exposure to pH 2.0 and was inactivated within 1 h at pH 4.0. The virus titer slowly declined, a 3-orders of magnitude reduction in virus titer, at pH 5.0 during a 4-h exposure. Virus survival at pH 6.0 was equal to that of the control in cell culture medium 199 MK (pH 7.12). PMID- 16347495 TI - Macromolecular Organization of the Cellulolytic Enzyme Complex of Clostridium thermocellum as Revealed by Electron Microscopy. AB - Clostridium thermocellum JW20 and YM4 both synthesize cellulolytic enzyme complexes, cellulosomes, when grown on medium containing cellulose. Electron microscopic studies showed that, in the early stages of growth of strain JW20, clusters of tightly packed cellulosomes, i.e., polycellulosomes, were located on the cell surface and were bound to cellulose. The polycellulosome was estimated to have a particle mass of 50 x 10 to 80 x 10 daltons (Da), while that of the cellulosome was estimated to be 2 x 10 to 2.5 x 10 Da and to contain about 35 polypeptides ranging from 20 to 200 kDa. The cellulosome produced by strain YM4 was found to be somewhat larger, with the estimated particle mass being 3.5 x 10 Da, and the number of polypeptides was counted to be 45 to 50, ranging from 20 to 200 kDa. In the early stages of cultivation, the cellulosomes from both species exist as tightly packed complexes (tight cellulosomes). These subsequently decompose to loosely packed complexes (loose cellulosomes) and ultimately to free polypeptides. Examination of the loose cellulosomal particles showed that they contain rows of equidistantly spaced, similarly sized polypeptide subunits, with an apparently identical orientation arranged parallel to the major axis of the cellulosome. It is postulated that on binding of a cellulose chain alongside such a row of subunits a simultaneous multicutting event occurs that leads to the release of cellooligosaccharides of four cellobiose units in length (C(4)). Rows of smaller-sized subunits with lower center-to-center distances, which are also present in the cellulosome, subsequently cleave the C(4) fragments (or cellulose) to C(2) (cellotetraose) or C(1) (cellobiose). In this way the cellulosome can catalyze the complete hydrolysis of cellulose. PMID- 16347496 TI - Identification and characterization of rhizosphere-competent bacteria of wheat. AB - To obtain rhizosphere-competent bacteria which could subsequently be modified for the development of biological control agents, bacteria were isolated from the rhizosphere and rhizoplane of wheat and barley plants by standard techniques. Of these isolates, 60 were selected for field testing as spring wheat seed inoculants in 1985. Isolates were marked genetically for resistance to antibiotics via selection of spontaneous mutants to detect and monitor isolates in the field. Forty-three days after planting, the average log(10) CFU/mg (dry weight) of roots and rhizosphere soil for the mutant isolates sampled ranged from 0 to 3.4. Twenty mutant isolates were retested in 1986. A total of 4 isolates were not detected, but the other 16 had an average root colonization value of log(10) 2.1 CFU and a range of log(10) 0.9 CFU to log(10) 3.2 CFU when sampled 32 days after planting. The average colonization value dropped to log(10) 1.1 CFU 51 days later. Some isolates detected previously were not detected in the second sampling; others had root colonization values similar to those obtained in the first sampling. Mutant isolates of rhizosphere bacteria included Bacillus pumilus, Bacillus subtilis, Pseudomonas fluorescens, Streptomyces spp., Xanthomonas maltophilia, and a saprophytic coryneform. Mixtures of isolates from different genera and species were compatible on seeds and roots. PMID- 16347497 TI - Production of Acetaldehyde by Zymomonas mobilis. AB - Mutants of Zymomonas mobilis were selected for decreased alcohol dehydrogenase activity by using consecutively higher concentrations of allyl alcohol. A mutant selected by using 100 mM allyl alcohol produced acetaldehyde at a level of 4.08 g/liter when the organism was grown in aerated batch cultures on a medium containing 4.0% (wt/wt) glucose. On the basis of the amount of glucose utilized, this level of acetaldehyde production represents nearly 40% of the maximum theoretical yield. Acetaldehyde produced during growth was continuously air stripped from the reactor. Acetaldehyde present in the exhaust stream was then trapped as the acetaldehyde-bisulfite addition product in an aqueous solution of sodium bisulfite and released by treatment with base. Acetaldehyde was found to inhibit growth of Z. mobilis at concentrations as low as 0.05% (wt/wt) acetaldehyde. An acetaldehyde-tolerant mutant of Z. mobilis was isolated after both mutagenesis with nitrosoguanidine and selection in the presence of vapor phase acetaldehyde. The production of acetaldehyde has potential advantages over that of ethanol: lower energy requirements for product separation, efficient separation of product from dilute feed streams, continuous separation of product from the reactor, and a higher marketplace value. PMID- 16347498 TI - Some Properties of Extracellular Acetylxylan Esterase Produced by the Yeast Rhodotorula mucilaginosa. AB - The red yeast Rhodotorula mucilaginosa produced an esterase that accumulated in the culture supernatant on induction with triacetin. The enzyme was specific for substrates bearing an O-acetyl group, but was relatively nonspecific for the rest of the molecule, which could consist of a phenol, a monosaccharide, a polysaccharide, or an aliphatic alcohol. The esterase was more active against acetylxylan and glucose beta-d-pentaacetate than were a number of esterases from plant and animal sources, when activities on 4-nitrophenyl acetate were compared. The enzyme exhibited Michaelis-Menten kinetics and was active over a broad pH range (5.5 to 9.2), with an optimum between pH 8 and 10. In addition, the enzyme retained its activity for 2 h at 55 degrees C. The yeast that produced the enzyme did not produce xylanase and, hence, is of interest for the production of acetylxylan esterase that is free of xylanolytic activity. PMID- 16347499 TI - Induction of Cellulolytic and Xylanolytic Enzyme Systems in Streptomyces spp. AB - Extracellular enzyme preparations from Streptomyces flavogriseus and Streptomyces olivochromogenes cultures grown on cellulose contained primarily cellulase activities, but similar preparations from cultures grown on xylan-containing materials possessed high levels of both cellulase and xylanase activities. Growth conditions that gave high endoxylanase levels also resulted in the production of enzymes involved in the hydrolysis of the nonxylose components of xylan. Specific acetyl xylan esterase activities were identified in enzyme preparations from both organisms. Both organisms also produced alpha-l-arabinofuranosidase activity that was not associated with endoxylanase activity. Other activities produced were alpha-l-O-methylglucuronidase and ferulic acid esterase. The latter enzyme was produced only by S. olivochromogenes and is an activity which has not previously been identified as a component of hemicellulase preparations. PMID- 16347500 TI - Cell-Free Solubilization of Coal by Polyporus versicolor. AB - Solubilization of coal was demonstrated with filtrates (0.45-mum-pore-size filters) obtained from the broth in which Polyporus versicolor had grown. The rate and extent of solubilization were dependent on the age of the fungal cultures, the particle size of the coal, the pH of the filtrates, and the presence of proteins in the filtrates. The rate of solubilization of coal was significantly reduced after proteins in the filtrates were denatured by acid hydrolysis. PMID- 16347501 TI - Solubilization of Leonardite by an Extracellular Fraction from Coriolus versicolor. AB - Coriolus versicolor has previously been shown to degrade leonardite, an oxidized form of lignite. An extracellular fraction containing protein purified from a C. versicolor culture solubilized leonardite in vitro. Expression of the activity did not require the presence of leonardite and appeared during idiophase. During ion-exchange and gel filtration column chromatography, leonardite-biosolubilizing activity eluted with syringaldazine oxidase activity and with protein, as measured by A(280) and the biuret protein assay. Syringaldazine is a substrate of the polyphenol oxidase formed by C. versicolor. Comparison of leonardite biosolubilizing activity with the effects of chelators and surface-active agents on leonardite showed that biosolubilization was not due to either surfactant or chelating ability. Heat treatment of the preparation at 60 degrees C for 30 min significantly reduced both syringaldazine oxidase and leonardite-biosolubilizing activities. Cyanide, azide, and thioglycolate, which are known inhibitors of syringaldazine oxidase activity of C. versicolor, also inhibited leonardite biosolubilization. From these data, we conclude that the purified protein fraction from C. versicolor contains a syringaldazine oxidase activity that participates in leonardite biosolubilization by enzymatic action. PMID- 16347502 TI - Effect of Butanol Challenge and Temperature on Lipid Composition and Membrane Fluidity of Butanol-Tolerant Clostridium acetobutylicum. AB - The effect of butanol challenge (0, 1.0, 1.5% [vol/vol]) and growth temperature (22, 37, 42 degrees C) on the membrane composition and fluidity of Clostridium acetobutylicum ATCC 824 and a butanol-tolerant mutant, SA-2, was examined in chemically defined medium. Growth of strain ATCC 824 into the stationary phase coincided with a gradual increase in the percent saturated to percent unsaturated (SU) fatty acid ratio. When challenged with butanol at 22 and 37 degrees C, ATCC 824 demonstrated an immediate (within 30 min) dose-response increase in the SU ratio. This strain showed little additional change over a 48-h fermentation. Compared with ATCC 824, growth of SA-2 into the late stationary phase at 22 or 37 degrees C resulted in an overall greater increase in the SU ratio for both unchallenged and challenged cells. This effect was minimized when SA-2 was challenged at 42 degrees C, probably due to the combination of the membrane fluidizing effect of butanol and the elevated temperature. Growth at 42 degrees C resulted in an increase in longer acyl chain fatty acids at the expense of shorter acyl chains for both strains. The membrane fluidity exhibited by SA-2 remained essentially constant at various butanol challenge and temperature combinations, while that for the ATCC 824 strain increased with increasing butanol challenge. By synthesizing an increased amount of saturated fatty acids, the butanol-tolerant SA-2 strain has apparently developed a mechanism for maintaining a more stable membrane environment. Growth of the microorganism is necessary for butanol to fluidize the membrane. Incorporation of exogenous fatty acids (18:1) did not significantly improve the butanol tolerance of either strain. Since SA-2 was able to produce only trace amounts of either butanol or acetone, increased tolerance to butanol does not necessarily coincide with greater solvent yields in this strain. PMID- 16347503 TI - Production rate of planktonic bacteria in the north basin of lake biwa, Japan. AB - Vertical and seasonal variations in the cell number and production rate of planktonic bacteria were investigated at a pelagic site (water depth, ca. 72 m) of the north basin of Lake Biwa during April to October 1986. The [methyl H]thymidine uptake rate into a cold trichloroacetic acid-insoluble fraction and the frequency of dividing cells (FDCs) were measured for each sample as indices of the bacterial production rate. The seasonal data of bacterial number, thymidine uptake rate, and bacterial growth rate based on the FDCs were correlated with one another (rank correlation analysis, P < 0.05). These bacterial variables were not correlated positively with the chlorophyll a concentration. Vertically, the maxima of both bacterial number and the thymidine uptake rate were found in the euphotic zone. The direct counting of bacteria and the measurements of thymidine uptake rate combined with the size-fractionation method revealed that more than 90% of the bacterial biomass and production rate were attributed to unattached bacteria throughout the investigation period. The carbon flux estimates of bacterial production were less certain due to the variability of the conversion factor for the thymidine uptake method and that of the calibration for the FDC method, but even when the conservative range of bacterial net production rate was used (5 to 60 mug of carbon per liter per day), it can be suggested that bacterial net production in the investigated area was a significant fraction (ca. 30%) of the level of the primary production rate in the same water basin. PMID- 16347504 TI - A Phosphate-Bond-Driven Dipeptide Transport System in Streptococcus cremoris Is Regulated by the Internal pH. AB - The uptake of amino acids and peptides by Streptococcus cremoris is mediated by different highly specific transport systems. The leucine transport system has a high affinity only for leucine, isoleucine, and valine and no affinity for leucyl peptides. The transport system for leucyl-leucine is strongly inhibited by several dipeptides with hydrophobic, neutral, N-terminal amino acids but not by leucine. The leucyl-leucine transport system has a high affinity for dipeptides containing beta-methyl groups in the side chain; the C terminus of the dipeptide affects the affinity to a much lower extent. Leucyl-leucine transport in whole cells was studied as a function of the internal pH at different external pH values in the presence and absence of nigericin. The internal pH was shown to be an important controlling factor in leucyl-leucine uptake, but the DeltapH was not involved as a driving force. At increasing external pH values, the affinity of the transport system for leucyl-leucine decreased. Uptake of leucyl-leucine was also studied in the presence of arsenate, which inhibited ATP synthesis by substrate-level phosphorylation. The rate of leucyl-leucine transport appeared to be dependent on the intracellular ATP concentrations. These results indicate that the energy for the leucyl-leucine transport is directly supplied by ATP. PMID- 16347505 TI - Oxidation of gaseous and volatile hydrocarbons by selected alkene-utilizing bacteria. AB - Eleven strains of alkene-utilizing bacteria belonging to the genera Mycobacterium, Nocardia, and Xanthobacter were tested for their ability to grow with C(1) to C(6) alkanes, C(2) to C(6) alkenes, alkadienes, and monoterpenes furnished individually as sole sources of carbon and energy in a mineral salts medium. A limited number of alkenes and alkanes supported growth of the bacteria; some bacteria were unable to grow on any of the saturated hydrocarbons tested. Monoterpenes were frequently used as carbon and energy sources by alkene utilizing bacteria belonging to the genera Mycobacterium and Nocardia. Washed cell suspensions of alkene-grown bacteria attacked the whole range of alkenes tested, whereas only three strains were able to oxidize alkanes as well. The alkenes tested were oxidized either to water and carbon dioxide or to epoxyalkanes. Few epoxides accumulated in stoichiometric amounts from the corresponding alkenes, because most epoxides formed were further converted to other compounds like alkanediols. PMID- 16347506 TI - Biosynthesis of Indole-3-Acetic Acid by the Pine Ectomycorrhizal Fungus Pisolithus tinctorius. AB - Previous work has indicated that anatomical and morphological changes (stunting and dichotomy) in roots of various conifers may be influenced by plant-growth regulating substances secreted by mycorrhizae. Indole-3-acetic acid (IAA) has been tentatively identified as a major auxin produced by some selected ectomycorrhizae. We report the isolation and detection of IAA as a secondary metabolite from Pisolithus tinctorius by thin-layer chromatography, high performance liquid chromatography (HPLC), enzyme-linked immunosorbent (monoclonal antibody) assay (ELISA), and unequivocal identification by gas chromatography mass spectrometry (GC-MS). The thin-layer chromatography methods for auxin isolation described here are novel, with the use of heptane-acetone-glacial acetic acid as the migrating solvent and formaldehyde, H(2)SO(4), and vanadate in detection. The acidic extract of the culture supernatant was methylated with ethereal diazomethane to detect IAA as methyl-3-IAA by HPLC, ELISA, and GC-MS. The quantitative amount of IAA detected ranged from 4 to 5 mumol liter by HPLC and ELISA. Another unidentified metabolite was detected by GC-MS with a typical indole nucleus (m/z = 130), indicating that it could be an intermediate in auxin metabolism. Plant response (Pseudotsuga menziesii, Douglas fir) was monitored upon inoculation of P. tinctorius and l-tryptophan. There was a consistent increase in plant height and stem diameter as a result of the two treatments, with statistical differences in dry weights of the shoots and roots. PMID- 16347507 TI - Tests of the critical assumptions of the dilution method for estimating bacterivory by microeucaryotes. AB - The critical assumptions of the dilution method for estimating grazing rates of microzooplankton were tested by using a community from the sediment-water interface of Lake Anna, Va. Determination of the appropriate computational model was achieved by regression analysis; the exponential model was appropriate for bacterial growth at Lake Anna. The assumption that the change in grazing pressure is linearly proportional to the dilution factor was tested by analysis of variance with a lack-of-fit test. There was a significant (P < 0.0001) linear (P > 0.05) relationship between the dilution factor and time-dependent change in ln bacterial abundance. The assumption that bacterial growth is not altered by possible substrate enrichment in the dilution treatment was tested by amending diluted water with various amounts of dissolved organic carbon (either yeast extract or extracted carbon from lake sediments). Additions of carbon did not significantly alter bacterial growth rates during the incubation period (24 h). On the basis of these results, the assumptions of the dilution method proved to be valid for the system examined. PMID- 16347508 TI - Calorimetric versus Growth Microbial Analysis of Cellulase Enzymes Acting on Cellulose. AB - ASSAY OF CELLULASE ENZYMOLOGY ON CELLULOSE WAS INVESTIGATED BY TWO METHODS: (i) plate colony counting to determine microbial growth and (ii) microbial calorimetry. These methods were chosen because they accept raw samples and have the potential to be far more specific than spectrophotometric reducing sugar assays. Microbial calorimetry requires ca. 0.5 to 1 h and 10 to 100 muM concentrations of cellulolytic lower sugars (glucose and cellobiose). Growth assay (liquid culture, plating, colony counting) requires 15 to 20 h and ca. 0.5 mM sugars. Microbial calorimetry requires simply aerobic metabolism, whereas growth assay requires completion of the cell cycle. A stripping technique is described for use in conjunction with the calorimetric method to enable separate analysis of the two sugars. Mixtures of glucose and cellobiose are equilibrated with Escherichia coli and spun out to remove glucose. The supernatant is calorimetrically combusted with Klebsiella sp. to quantitate cellobiose, and the same organism combusting the nonstripped mixture gives heat proportional to the sum of the two sugars. Calorimetry of cellulolysis products from individual exo- and endocellulases, and from their reconstituted mixture, was carried out to develop a microbial calorimetric means for demonstrating enzyme synergism. PMID- 16347509 TI - Evidence for genetic exchange and recombination of Rhizobium symbiotic plasmids in a soil population. AB - A soil population of 16 Rhizobium leguminosarum bv. trifolii isolates was characterized by using three Sym (for symbiotic) plasmid-specific DNA hybridization probes: (i) an R. leguminosarum bv. trifolii-specific, repeated sequence probe; (ii) a nifHDK gene probe, and (iii) a nod gene probe. A predominant Sym plasmid family was identified among the isolates. Three other unrelated Sym plasmid families were also identified. The isolates were also classified either by using a chromosomal DNA hybridization probe or by serological relatedness to 25 different R. leguminosarum bv. trifolii antisera. With either method, it was possible to group the 16 soil isolates into identical or related families. However, the correlation between the two techniques was not high. Irrespective of the means used to classify the bacterial host strain, it was possible to identify the same Sym plasmids in unrelated strains, as well as unrelated Sym plasmids in identical host strains. These data indicate that, within this soil population, there has been genetic exchange of Sym plasmids, and in one instance the hybridization pattern indicates that in vivo recombination of two different Sym plasmids may have occurred. Symbiotic effectiveness tests on red, strawberry, and subterranean clovers clearly differentiated the isolates. In general, the pattern of response was similar within groupings on the basis of Sym plasmid and chromosomal profiles but different between such groups. PMID- 16347510 TI - Chelating Properties of Extracellular Polysaccharides from Chlorella spp. AB - Chlorella stigmatophora LB 993 was grown in artificial seawater under controlled conditions. The production of cell wall polysaccharides attached to the cells and dissolved in the growth medium was monitored during algal growth. Preliminary characterization of the dissolved polysaccharides of C. stigmatophora and other Chlorella species is presented. The capacity of dissolved polysaccharides of C. stigmatophora to bind toxic heavy metals was also studied and compared with that of polysaccharides produced by other marine Chlorella species. The differences in metal-complexing capacity observed for dissolved polysaccharides obtained from various Chlorella species is attributable to differences in the composition of the polysaccharides, notably the uronic acids content. PMID- 16347511 TI - Characteristics of Cyclic AMP Transport by Marine Bacteria. AB - Uptake and autoradiography experiments with natural populations of marine bacteria, seawater cultures, and cultured isolates showed that the high-affinity cyclic AMP transport system in marine bacteria has stringent structural requirements, is found in a minority of cells in mixed bacterial assemblages, and appears to be related to the culture growth state. PMID- 16347512 TI - Variability in Effectiveness of Rhizobia during Culture and in Nodules. AB - The ability of three strains of Bradyrhizobium sp. (Vigna) to fix dinitrogen in symbiotic association with siratro (Macropitilium atropurpureum) was measured after culture in broth and after isolation from nodules. Seven transfers were made between the initial broth culture and the final broth culture. A total of 40 single-colony isolates were obtained from cultures 1 and 7 to test effectiveness. Variation in dinitrogen-fixing effectiveness of the population of one strain did not change on culturing, whereas there was considerable variation in effectiveness of populations of the other two strains. Generally, single-colony isolates from individual nodules had similar levels of effectiveness, but some exceptions occurred. Isolates from different nodules formed by the same Bradyrhizobium strain often differed in their effectiveness. PMID- 16347513 TI - Cloning of the Pachysolen tannophilus Xylulokinase Gene by Complementation in Escherichia coli. AB - The gene coding for xylulokinase has been isolated from the yeast Pachysolen tannophilus by complementation of Escherichia coli xylulokinase (xylB) mutants. Through subcloning, the gene has been localized at one end of a 3.2-kilobase EcoRI-PstI fragment. Expression of the cloned gene was insensitive to glucose inhibition. Furthermore, the cloned gene did not cross-hybridize with E. coli and Saccharomyces cerevisiae xylulokinase genes. PMID- 16347514 TI - Factors Affecting the Methanogenic Activity of Methanothrix soehngenii VNBF. AB - Methane production by Methanothrix soehngenii VNBF grown on acetate (50 mM) as the sole carbon and energy source was influenced by the addition of Fe, trace elements, and pesticides. The addition of Fe and trace elements significantly enhanced the rate of CH(4) production. The addition of pesticides in the early growth phase caused complete inhibition. However, less inhibition was noted when pesticides were added during early exponential growth phase. Addition to culture tubes of Co, Ni, or Mo at 2 muM produced 64, 41, or 17%, respectively, more CH(4) than that produced in tubes lacking the corresponding trace element. A concentration of more than 5 muM of these trace elements in the medium resulted in decreased CH(4) production, presumably because of toxic effects. PMID- 16347515 TI - Properties of Lactose Plasmid pLY101 in Lactobacillus casei. AB - A starter strain, Lactobacillus casei C257, was found to carry a lactose plasmid, pLY101. Restriction mapping showed that pLY101 DNA was 68.2 kilobases long. Since a non-lactose-utilizing variant of C257, MSK248, lost phospho-beta-galactosidase (P-beta-gal) activity and pLY101 DNA had a sequence(s) homologous to the streptococcal fragment including a P-beta-gal gene, pLY101 is likely to encode a P-beta-gal gene required for lactose metabolism in C257. MSK248 grew in galactose medium at a rate identical to that of C257 and retained phosphoenolpyruvate dependent phosphotransferase system activity for lactose similar to that of C257. Therefore, the C257 chromosome appears to encode a complete set of genes for the lactose-phosphotransferase system and the predominant galactose metabolic pathway in C257. pLY101 DNA had a sequence homologous to a lactobacillus insertion sequence, ISL1, which mapped more than 12 kilobases from the sequence homologous to the streptococcal P-beta-gal fragment. PMID- 16347516 TI - Influence of physical disruption on growth of attached bacteria. AB - Attached bacterial populations cultured without an exogenous carbon source or grown in conjunction with attached diatoms incorporated [H]thymidine at a rate between four and five times lower than that of replicate bacterial populations which were dispersed before being assayed. PMID- 16347517 TI - Control of Interspecies Electron Flow during Anaerobic Digestion: Role of Floc Formation in Syntrophic Methanogenesis. AB - The flora of an anaerobic whey-processing chemostat was separated by anaerobic sedimentation techniques into a free-living bacterial fraction and a bacterial floc fraction. The floc fraction constituted a major part (i.e., 57% total protein) of the total microbial population in the digestor, and it accounted for 87% of the total CO(2)-dependent methanogenic activity and 76% of the total ethanol-consuming acetogenic activity. Lactose was degraded by both cellular fractions, but in the free flora fraction it was associated with higher intermediary levels of H(2), ethanol, butyrate, and propionate production. Electron microscopic analysis of flocs showed bacterial diversity and juxtapositioning of tentative Desulfovibrio and Methanobacterium species without significant microcolony formation. Ethanol, an intermediary product of lactose hydrolyzing bacteria, was converted to acetate and methane within the flocs by interspecies electron transfer. Ethanol-dependent methane formation was compartmentalized and closely coupled kinetically within the flocs but without significant formation of H(2) gas. Physical disruption of flocs into fragments of 10- to 20-mum diameter initially increased the H(2) partial pressure but did not change the carbon transformation kinetic patterns of ethanol metabolism or demonstrate a significant role for H(2) in CO(2) reduction to methane. The data demonstrate that floc formation in a whey-processing anaerobic digestor functions in juxtapositioning cells for interspecies electron transfer during syntrophic ethanol conversion into acetate and methane but by a mechanism which was independent of the available dissolved H(2) gas pool in the ecosystem. PMID- 16347518 TI - Isolation and Characterization of a Thermophilic Bacterium Which Oxidizes Acetate in Syntrophic Association with a Methanogen and Which Grows Acetogenically on H(2)-CO(2). AB - We previously described a thermophilic (60 degrees C), syntrophic, two-membered culture which converted acetate to methane via a two-step mechanism in which acetate was oxidized to H(2) and CO(2). While the hydrogenotrophic methanogen Methanobacterium sp. strain THF in the biculture was readily isolated, we were unable to find a substrate that was suitable for isolation of the acetate oxidizing member of the biculture. In this study, we found that the biculture grew on ethylene glycol, and an acetate-oxidizing, rod-shaped bacterium (AOR) was isolated from the biculture by dilution into medium containing ethylene glycol as the growth substrate. When the axenic culture of the AOR was recombined with a pure culture of Methanobacterium sp. strain THF, the reconstituted biculture grew on acetate and converted it to CH(4). The AOR used ethylene glycol, 1,2 propanediol, formate, pyruvate, glycine-betaine, and H(2)-CO(2) as growth substrates. Acetate was the major fermentation product detected from these substrates, except for 1,2-propanediol, which was converted to 1-propanol and propionate. N,N-Dimethylglycine was also formed from glycine-betaine. Acetate was formed in stoichiometric amounts during growth on H(2)-CO(2), demonstrating that the AOR is an acetogen. This reaction, which was carried out by the pure culture of the AOR in the presence of high partial pressures of H(2), was the reverse of the acetate oxidation reaction carried out by the AOR when hydrogen partial pressures were kept low by coculturing it with Methanobacterium sp. strain THF. The DNA base composition of the AOR was 47 mol% guanine plus cytosine, and no cytochromes were detected. PMID- 16347519 TI - Methanogenesis from Methylated Amines in a Hypersaline Algal Mat. AB - Methane ebullition and high rates of methane production were observed in sediments of a hypersaline pond (180 per thousand) which contained sulfate in excess of 100 mM. The highest rates of methane production were observed in surface sediments associated with an algal mat dominated by a Spirulina sp. The mat contained a methylated amine, glycine betaine (GBT), at levels which accounted for up to 20% of the total mat nitrogen. GBT was apparently the source of trimethylamine (TMA), which was also present in the sediment at relatively high concentrations. Patterns of substrate metabolism by the methanogenic populations in sediment slurries suggested that TMA was a major methane precursor. Neither exogenous hydrogen nor acetate stimulated methanogenesis, while addition of a variety of amines including TMA, trimethylamine oxide, GBT, and choline resulted in substantial increases with yields of >70%. The temperature optimum for methanogenesis in this system was 45 to 55 degrees C, which coincided with the observed sediment temperature. Patterns and rates of methane production in this and other hypersaline algal mats may be determined by a complex interaction between salinity, the use of methylated amines for osmoregulation by algae, and the formation of TMA by fermentation. PMID- 16347520 TI - Acetic Acid Production by an Electrodialysis Fermentation Method with a Computerized Control System. AB - In acetic acid fermentation by Acetobacter aceti, the acetic acid produced inhibits the production of acetic acid by this microorganism. To alleviate this inhibitory effect, we developed an electrodialysis fermentation method such that acetic acid is continuously removed from the broth. The fermentation unit has a computerized system for the control of the pH and the concentration of ethanol in the fermentation broth. The electrodialysis fermentation system resulted in improved cell growth and higher productivity over an extended period; the productivity exceeded that from non-pH-controlled fermentation. During electrodialysis fermentation in our system, 97.6 g of acetic acid was produced from 86.0 g of ethanol; the amount of acetic acid was about 2.4 times greater than that produced by non-pH-controlled fermentation (40.1 g of acetic acid produced from 33.8 g of ethanol). Maximum productivity of electrodialysis fermentation in our system was 2.13 g/h, a rate which was 1.35 times higher than that of non-pH-controlled fermentation (1.58 g/h). PMID- 16347521 TI - Synthesis of an Iron-Oxidizing System during Growth of Thiobacillus ferrooxidans on Sulfur-Basal Salts Medium. AB - It was found that the de novo synthesis of not only sulfur:ferric ion oxidoreductase (ferric ion-reducing system) but also iron oxidase was absolutely required when Thiobacillus ferrooxidans AP19-3 was grown on sulfur-salts medium. The results strongly suggest that iron oxidase is involved in sulfur oxidation. This bacterium could not grow on sulfur-salts medium under anaerobic conditions with Fe as a terminal electron acceptor, suggesting that energy conservation by electron transfer between elemental sulfur and Fe is not available for this bacterium. PMID- 16347522 TI - Development of a Mutant Strain of Bacillus polymyxa Showing Enhanced Production of 2,3-Butanediol. AB - 2,3-Butanediol is a feedstock chemical of potential industrial importance. It can serve as a monomer for many polymers of consumer interest that are currently supplied by the fossil fuel industry. Bacillus polymyxa can grow on inexpensive waste products of the food-processing industry and produce this glycol. This paper describes a mutant strain of B. polymyxa which displays constitutive production of catabolic alpha-acetolactate synthase, an enzyme in the 2,3 butanediol pathway which is normally produced only in the late log or stationary phase of growth. The mutant was obtained by treating the wild type with nitrosoguanidine and subjecting it to a penicillin counterselection procedure. One of the selected mutant strains produced four times as much of the glycol as the wild type and utilized approximately 25% of the energy source, compared with essentially complete utilization of the energy source by the wild type. Studies are under way to optimize the production of the glycol by the mutant. PMID- 16347523 TI - Photosynthetic Potential and Light-Dependent Oxygen Consumption in a Benthic Cyanobacterial Mat. AB - The potential to carry out oxygenic photosynthesis after prolonged burial below the photic zone was studied at 0.1-mm depth intervals in the thick, laminated Microcoleus chthonoplastes mats growing in Solar Lake, Sinai. The buried mat community lost about 20% of its photosynthetic potential with depth per annual layer down to 8- to 10-year-old layers at a 14-mm depth. In some of the older layers, below a 30-mm depth, light-dependent oxygen consumption which increased with increasing light intensity was observed. Possible mechanisms for this phenomenon are (i) pseudocyclic electron transport (Mehler reaction), (ii) interactions between respiratory electron transport and photosynthetic electron transport, (iii) photorespiration, and (iv) photooxidation. PMID- 16347524 TI - High Survivability of Cheese Whey-Grown Rhizobium meliloti Cells upon Exposure to Physical Stress. AB - Whey, a by-product of the dairy industry, has been found to protect the rhizobia cells during freezing and thawing. Cells of rhizobia grown on whey sustained freezing better at -18 degrees C than did cells grown on mannitol or sucrose. Suspensions of cells grown on whey or mannitol that were suspended in whey performed equally well at -18 and -80 degrees C, with 94 and 100% survival, respectively. Whey-grown rhizobia in pellets withstood desiccation better than did their mannitol-grown equivalents. Rhizobia that were grown on whey and then inoculated onto commercial peat showed a survival rate of 100% after 23 weeks at 4 degrees C. Whey-grown cells in peat performed better at various temperatures during storage, even when they were exposed to desiccation, than did mannitol grown cells in peat. Whey, therefore, offers interesting possibilities as a Rhizobium protectant for the inoculum industry. PMID- 16347525 TI - A Study of 33 Bacteriophages of Rhizobium meliloti. AB - A total of 33 Rhizobium meliloti bacteriophages were studied. Of those, 21 were isolated in northern France from field soil in which Medicago sativa L. was grown. The other 12 phages were obtained by UV light and mitomycin C induction from 46 R. meliloti strains. Rhizobiophages were characterized by their morphology, host range, serological properties, restriction endonuclease patterns, DNA-DNA homologies, and DNA molecular weights. Five morphotypes were observed showing tailed phages with icosahedral heads. The categories of morphotypes included the Myoviridae (11 phages), Siphoviridae (3 morphotypes and 20 phages), and Podoviridae (2 phages). Type NM1 phage (Siphoviridae) is highly unusual because of the presence of transverse bars on the phage tail. Soil phages had broad host ranges, whereas phages isolated from bacterial cultures showed more or less narrow host ranges. Restriction endonuclease patterns and DNA-DNA hybridization experiments showed that the five phage type genomes were unrelated. Molecular weights of phage type DNAs were estimated, and they corresponded to values expected for capsid sizes, except for phage NM8. Type phiM11S (Siphoviridae) did not correspond to any other described Rhizobium phages and represents a new species. PMID- 16347526 TI - Control of Interspecies Electron Flow during Anaerobic Digestion: Significance of Formate Transfer versus Hydrogen Transfer during Syntrophic Methanogenesis in Flocs. AB - Microbial formate production and consumption during syntrophic conversion of ethanol or lactate to methane was examined in purified flocs and digestor contents obtained from a whey-processing digestor. Formate production by digestor contents or purified digestor flocs was dependent on CO(2) and either ethanol or lactate but not H(2) gas as an electron donor. During syntrophic methanogenesis, flocs were the primary site for formate production via ethanol-dependent CO(2) reduction, with a formate production rate and methanogenic turnover constant of 660 muM/h and 0.044/min, respectively. Floc preparations accumulated fourfold higher levels of formate (40 muM) than digestor contents, and the free flora was the primary site for formate cleavage to CO(2) and H(2) (90 muM formate per h). Inhibition of methanogenesis by CHCl(3) resulted in formate accumulation and suppression of syntrophic ethanol oxidation. H(2) gas was an insignificant intermediary metabolite of syntrophic ethanol conversion by flocs, and its exogenous addition neither stimulated methanogenesis nor inhibited the initial rate of ethanol oxidation. These results demonstrated that >90% of the syntrophic ethanol conversion to methane by mixed cultures containing primarily Desulfovibrio vulgaris and Methanobacterium formicicum was mediated via interspecies formate transfer and that <10% was mediated via interspecies H(2) transfer. The results are discussed in relation to biochemical thermodynamics. A model is presented which describes the dynamics of a bicarbonate-formate electron shuttle mechanism for control of carbon and electron flow during syntrophic methanogenesis and provides a novel mechanism for energy conservation by syntrophic acetogens. PMID- 16347527 TI - Characterization of Clostridium thermocellum JW20. AB - Clostridium thermocellum JW20 (ATCC 31549), which was isolated from a Louisiana cotton bale, grew on cellulose, cellobiose, and xylooligomers and, after adaptation, on glucose, fructose, and xylose in the pH range of 7.5 to 6.1 with T(opt) of 60 degrees C, T(max) of 69 degrees C, and T(min) of above 28 degrees C. Doubling times during growth on cellulose and cellobiose were 6.5 and 2.5 h, respectively. The G+C content of the DNA was 40 mol% (chemical analysis). Growth on cellulose as substrate was totally inhibited in the presence of more than 125 mM sodium sulfate, 300 mM sodium chloride, 250 mM potassium chloride, 200 mM calcium chloride, 125 mM magnesium chloride, 40 mM lactate, or 250 mM acetate. The ratio of the fermentation products ethanol to acetate plus H(2) decreased when the culture was agitated. Agitation otherwise increased the rate of cellulose degradation in a growing culture but not under nongrowth conditions or with cell-free culture supernatant containing the extracellular cellulase. Shaking lowered the concentration of H(2) in the culture broth and thus minimized inhibition by the H(2) formed. Externally added H(2) caused an increased formation of ethanol during growth on cellulose or cellobiose. However, at an atmospheric pressure as high as 355 kPa (50 lb/in), H(2) did not cause significant growth inhibition beyond an increasing lag phase (up to 24 h). Several criteria to specifically prove the purity of C. thermocellum cultures were suggested. PMID- 16347528 TI - Effects of Cellulolytic Ruminal Bacteria and of Cell Extracts on Germination of Euonymus americanus L. Seeds. AB - In past attempts, the experimental germination of the seeds of Euonymus americanus L. in vitro has had little success. However, treatment of seeds with ruminal fluid containing viable microflora has been successful in stimulating germination. In the presence of the cellulolytic ruminal bacterium, Clostridium cellobioparum ATCC 15832, seeds of E. americanus were stimulated to germinate. Subsequent studies were designed to determine whether the bacterium synthesized a cellulolytic enzyme responsible for initiating germination. The cell-free endocellulase from C. cellobioparum induced germination of the seeds. To support the hypothesis that the endocellulase from C. cellobioparum was responsible for triggering germination, a 1,4-beta-d-glucan glucanohydrolase (EC 3.2.1.4) from Penicillum funiculosum was used to treat the seeds. In addition, no germination was obtained from seeds treated with a commercial exocellulase enzyme. Also, Ruminococcus flavefaciens FD-1 was found to initiate germination of E. americanus seeds. Thus, cellulase activity is indicated in the degradation of the testa of the seed, allowing imbibition and germination. PMID- 16347529 TI - Construction of a Stable alpha-Galactosidase-Producing Baker's Yeast Strain. AB - Molasses is widely used as a substrate for commercial yeast production. The complete hydrolysis of raffinose, which is present in beet molasses, by Saccharomyces strains requires the secretion of alpha-galactosidase, in addition to the secretion of invertase. Raffinose is not completely utilized by commercially available yeast strains used for baking, which are Mel. In this study we integrated the yeast MEL1 gene, which codes for alpha-galactosidase, into a commercial mel baker's yeast strain. The Mel phenotype of the new strain was stable. The MEL1 gene was expressed when the new Mel baker's yeast was grown in molasses medium under conditions similar to those used for baker's yeast production at commercial factories. The alpha-galactosidase produced by this novel baker's yeast strain hydrolyzed all the melibiose that normally accumulates in the growth medium. As a consequence, additional carbohydrate was available to the yeasts for growth. The new strain also produced considerably more alpha galactosidase than did a wild-type Mel strain and may prove useful for commercial production of alpha-galactosidase. PMID- 16347530 TI - Growth of Indigenous Rhizobium leguminosarum and Rhizobium meliloti in Soils Amended with Organic Nutrients. AB - The ability of indigenous Rhizobium leguminosarum and Rhizobium meliloti to use organic nutrients as growth substrates in soil was assessed by indirect bacteriophage analysis. A total of 17 organic compounds, including 9 carbohydrates, 3 organic acids, and 5 amino acids, were tested (1,000 mug g) in three soils with different cropping histories. Four additional soils were screened with a glucose amendment. Nutrient amendments stimulated growth of indigenous rhizobia, allowing subsequent replication of indigenous bacteriophages. Phage populations were enumerated by plating soil extracts on 19 R. leguminosarum and 9 R. meliloti indicator strains, including root nodule isolates from the soils assayed. On the basis of indirect phage analysis, all soils contained native rhizobia similar to one or more of the indicator strains, although not all indicator strains were detected in soil. All organic compounds stimulated growth of indigenous rhizobia, but the growth response varied for each rhizobial strain depending on the nutrient, the nutrient concentration, and the soil. Indigenous rhizobia readily utilized most organic compounds except phenylalanine, glycine, and aspartic acid. The ability of indigenous rhizobia to utilize a wide range of organic compounds as growth substrates in situ indicates their ability to successfully compete with other soil bacteria for nutrients in these soils. PMID- 16347532 TI - Induction and Characterization of Artificial Diploids from the Haploid Yeast Torulaspora delbrueckii. AB - [This corrects the article on p. 1508 in vol. 53.]. PMID- 16347531 TI - Isolation of Auxotrophic Mutants of Methylophilus methylotrophus by Modified Marker Exchange. AB - A method for stabilizing a transposon (Tn5) has been developed which allows the isolation of stable auxotrophic mutants of Methylophilus methylotrophus ASI. Insertion of Tn5 into a cloned M. methylotrophus ASI DNA fragment encoding anthranilate synthase followed by transfer of the vector with the modified trpE gene to M. methylotrophus ASI resulted in unstable auxotrophs among the recombinants. Deletion of IS50R, which encodes transposase production from Tn5, stabilized the transposon after mobilization to M. methylotrophus ASI. When trpE genes with the modified Tn5 inserts were mobilized into M. methylotrophus ASI, stable, kanamycin-resistant tryptophan auxotrophs were obtained by double crossover homologous recombination with the chromosome. PMID- 16347534 TI - New Selective and Differential Medium for Vibrio cholerae and Vibrio vulnificus. AB - [This corrects the article on p. 2263 in vol. 53.]. PMID- 16347533 TI - Industrial Application of Artificially Induced Diploid Strains of Torulaspora delbrueckii. AB - [This corrects the article on p. 1512 in vol. 53.]. PMID- 16347535 TI - Metabolism of Lignin Model Compounds of the Arylglycerol-beta-Aryl Ether Type by Pseudomonas acidovorans D(3). AB - [This corrects the article on p. 2608 in vol. 53.]. PMID- 16347536 TI - Cloning and Gene Replacement Mutagenesis of a Pseudomonas atlantica Agarase Gene. AB - An agarase gene (agrA) was isolated by cloning genomic DNA prepared from Pseudomonas atlantica. The agarase activity in recombinant Escherichia coli was found in cell-free culture supernatants and could pass through a 0.45-mum-pore size membrane separating cells from agar, suggesting that the gene product was exported in E. coli. The enzyme was specific for agar and agarose and did not digest alginate or carrageenan. Mutations generated by transposon mini-Mu d1(lacZ Km) were used to define the agrA coding region, as well as the direction of transcription of the gene. A procedure was developed to produce a P. atlantica agrA mutant. This required construction of an agrA::kan insertion mutation in vitro and subsequent introduction of the defect into the chromosome of P. atlantica by recombinational exchange. Transformation of P. atlantica with plasmids containing agrA::kan utilized a Tris-polyethylene glycol 6000-CaCl(2) treatment for making competent cells. Replacement of wild-type agrA with agrA::kan resulted in loss of agarase activity. Uses of the agrA gene probe and an Agr mutant for environmental studies are discussed. PMID- 16347537 TI - Purification and Characterization of a Dipeptidase from Streptococcus cremoris Wg2. AB - A dipeptidase was purified to homogeneity from a crude cell extract of Streptococcus cremoris Wg2 by DEAE-Sephacel column chromatography followed by preparative disc gel electrophoresis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed a single protein band with a molecular weight of 49,000. The dipeptidase is capable of hydrolyzing a range of dipeptides, but not peptides with longer chains. The enzyme was shown to be a metallo-Mn enzyme with a pH optimum of 8 and a temperature optimum of 50 degrees C. The enzyme is strongly inhibited by thiol-reducing reagents but not by sulfhydryl reagents. Kinetic studies indicated that the enzyme has a relatively low affinity for leucyl-leucine and alanyl-alanine (K(m), 1.6 and 7.9 mM, respectively) but can hydrolyze these substrates at very high rates (V(max), 3,700 and 13,000 mumol/min per mg of protein, respectively). PMID- 16347538 TI - Induction of Xylose Reductase and Xylitol Dehydrogenase Activities in Pachysolen tannophilus and Pichia stipitis on Mixed Sugars. AB - The induction of xylose reductase and xylitol dehydrogenase activities on mixed sugars was investigated in the yeasts Pachysolen tannophilus and Pichia stipitis. Enzyme activities induced on d-xylose served as the controls. In both yeasts, d glucose, d-mannose, and 2-deoxyglucose inhibited enzyme induction by d-xylose to various degrees. Cellobiose, l-arabinose, and d-galactose were not inhibitory. In liquid batch culture, P. tannophilus utilized d-glucose and d-mannose rapidly and preferentially over d-xylose, while d-galactose consumption was poor and lagged behind that of the pentose sugar. In P. stipitis, all three hexoses were used preferentially over d-xylose. The results showed that the repressibility of xylose reductase and xylitol dehydrogenase may limit the potential of yeast fermentation of pentose sugars in hydrolysates of lignocellulosic substrates. PMID- 16347539 TI - Cloning and Mapping of a Novel Nodulation Region from Bradyrhizobium japonicum by Genetic Complementation of a Deletion Mutant. AB - The phenotypes of a set of Bradyrhizobium japonicum 110 mutants with large deletions in the region of symbiotic gene cluster I were tested. The majority of the mutants showed a delayed nodulation on soybean and, by mixed-infection experiments, were found to be strongly reduced in their competitiveness. Phenotypic comparison of mutants with different deletion endpoints allowed a preliminary localization of two genomic regions, called nod-1 and nod-2, which were required for normal nodulation on soybean. Loss of nod-1 was found to result in a Nod phenotype on cowpea, mung bean, and siratro. A recombinant cosmid was identified which fully restored nodulation ability of a mutant lacking nod-1. Using Tn5-containing derivatives and subclones of this cosmid for complementation, we delimited the nod-1 region to a DNA segment of 3.1 to 3.5 kilobase pairs. PMID- 16347540 TI - Delignification of Wood Chips and Pulps by Using Natural and Synthetic Porphyrins: Models of Fungal Decay. AB - Kraft pulps, prepared from softwoods, and small chips of birch wood were treated with heme and tert-butyl hydroperoxide in aqueous solutions at reflux temperature. Analyses of treated pulps showed decreases in kappa number (a measure of lignin content) from about 36 to less than 2, with concomitant increases in brightness (80% increase in the better samples). Analyses of treated wood chips revealed selective delignification and removal of hemicelluloses. After 48 h of treatment, lignin losses from the wood chips approached 40%, and xylose/mannose (hemicellulose) losses approached 70%, while glucose (cellulose) losses were less than 10%. Examination of delignified chips by transmission electron microscopy showed that the removal of lignin occurred in a manner virtually indistinguishable from that seen after decay by white rot fungi. Various metalloporphyrins, which act as biomimetic catalysts, were compared to horseradish peroxidase and fungal manganese peroxidase in their abilities to oxidize syringaldazine in an organic solvent, dioxane. The metalloporphyrins and peroxidases behaved similarly, and it appeared that the activities of the peroxidases resulted from the extraction of heme into the organic phase, rather than from the activities of the enzymes themselves. We concluded that heme-tert butyl hydroperoxide systems in the absence of a protein carrier mimic the decay of lignified tissues by white rot fungi. PMID- 16347541 TI - Quantitative immunologic analysis of the methanogenic flora of digestors reveals a considerable diversity. AB - To determine which methanogens occur in digestors, we performed a quantitative immunologic analysis of a variety of samples. A comprehensive panel of calibrated polyclonal antibody probes of predefined specificity spectra was used. This allowed precise identification of bacteria by antigenic fingerprinting. A considerable diversity of methanogens was uncovered, much larger than previously reported, encompassing at least 14 strains of 11 species. Strategies were developed to measure the load of any given methanogen in a sample and to compare samples quantitatively. Two methanogens were found to predominate which were antigenically closely related with either Methanobacterium formicicum MF or Methanobrevibacter arboriphilus AZ. Fundamental data, probes, and methods are now available to monitor methanogenic subpopulations during digestor operation and thus learn about their respective roles and predictive significance. PMID- 16347542 TI - Nodulation, Nitrogen Fixation, and Hydrogen Oxidation by Pigeon Pea Bradyrhizobium spp. in Symbiotic Association with Pigeon Pea, Cowpea, and Soybean. AB - The pigeon pea strains of Bradyrhizobium CC-1, CC-8, UASGR(S), and F4 were evaluated for nodulation, effectiveness for N(2) fixation, and H(2) oxidation with homologous and nonhomologous host plants. Strain CC-1 nodulated Macroptilium atropurpureum, Vigna unguiculata, Glycine max, and G. soja but did not nodulate Pisum sativum, Phaseolus vulgaris, Trigonella foenum-graecum, and Trifolium repens. Strain F4 nodulated G. max cv. Peking and PI 434937 (Malayan), but the symbioses formed were poor. Similarly, G. max cv. Peking, cv. Bragg, PI 434937, PR 13-28-2-8-7, and HM-1 were nodulated by strain CC-1, and symbioses were also poor. G. max cv. Williams and cv. Clark were not nodulated. H(2) uptake activity was expressed with pigeon pea and cowpea, but not with soybean. G. max cv. Bragg grown in Bangalore, India, in local soil not previously exposed to Bradyrhizobium japonicum formed nodules with indigenous Bradyrhizobium spp. Six randomly chosen isolates, each originating from a different nodule, formed effective symbioses with pigeon pea host ICPL-407, nodulated PR 13-28-2-8-7 soybean forming moderately effective symbioses, and did not nodulate Williams soybean. These results indicate the six isolates to be pigeon pea strains although they originated from soybean nodules. Host-determined nodulation of soybean by pigeon pea Bradyrhizobium spp. may depend upon the ancestral backgrounds of the cultivars. The poor symbioses formed by the pigeon pea strains with soybean indicate that this crop should be inoculated with B. japonicum for its cultivation in soils containing only pigeon pea Bradyrhizobium spp. PMID- 16347543 TI - Characterization of an Endoglucanase from Pseudomonas fluorescens subsp. cellulosa Produced in Escherichia coli and Regulation of the Expression of Its Cloned Gene. AB - Several enzymatic properties of an endoglucanase produced in Escherichia coli by a gene from Pseudomonas fluorescens subsp. cellulosa were investigated. Gel filtration revealed a single peak of M(r) 36,000 with endoglucanase activity. The pH optimum of the enzyme was 7.0. Carboxymethyl cellulose and barley beta-glucan (mixed beta-1,3 and 1,4 linkages) were good substrates, but not laminarin (beta 1,3 linkages), amylose, filter paper, microcrystalline cellulose (Avicel), or cellotriose. The mode of action was typical of an "endo"-acting enzyme. Taken together, these properties do not correspond to those of any of the endoglucanases described in P. fluorescens subsp. cellulosa. Consequently, the gene was designated egIX. The enzyme was sensitive to end-product inhibition by cellobiose but was only moderately inhibited by glucose. The enzyme was formed constitutively in E. coli throughout the growth phase. Urea had no effect on endoglucanase synthesis, but glucose acted as a catabolite repressor. The formation of the enzyme in E. coli was partially dependent on cyclic AMP. PMID- 16347544 TI - Production of Biodispersan by Acinetobacter calcoaceticus A2. AB - Bacterial strains A2 and HE5, isolated by enrichment culture techniques, were shown to produce extracellular, nondialyzable materials which disperse limestone powders in water. These materials are referred to as biodispersans. Strains A2 and HE5 were classified as Acinetobacter calcoaceticus by physiological and genetic tests. An assay for limestone-dispersing activity was developed which is based on the settling time of a standard 10% limestone-in-water mixture. The assay was proportional to biodispersan concentration between 40 and 100 mug/ml. Dispersion was optimum between pH 9 and 12. Phosphate (2 mM) and magnesium (8 mM) ions caused a 50% inhibition of activity. An optimized medium for the production of biodispersan was developed with ethanol as the substrate. Biodispersan was produced only after the bacteria completed their exponential growth phase and continued during the stationary phase, reaching over 4 g of crude biodispersan per liter. PMID- 16347545 TI - Purification and Chemical Properties of Acinetobacter calcoaceticus A2 Biodispersan. AB - The extracellular dispersant of Acinetobacter calcoaceticus A2, referred to as biodispersan, was concentrated by ammonium sulfate precipitation and deproteinized by hot phenol treatment. The active component was an anionic polysaccharide (PS-A2). The specific activity of PS-A2 was approximately three times greater than that of crude biodispersan. PS-A2 had a sedimentation constant of 1.39 S, a diffusion coefficient of 18.8 x 10 cm s, and a partial molar volume of 0.65 cm g, yielding an average molecular weight of 51,400. Titration of the polymer gave two inflection points: pK(1) = 3.1 (1.15 muEq/mg) and pK(2) = 8.0 (0.4 muEq/mg). PS-A2 slowly consumed 1.10 mumol of periodate per mg. The C nuclear magnetic resonance spectrum of PS-A2 indicated four methyl groups, four carbonyl C atoms, and four signals in the anomeric region (95 to 110 ppm), indicative of the presence of four different monosaccharides. Strong acid hydrolysis of PS-A2 yielded four reducing sugars: glucosamine, a 6-methyl aminohexose, galactosamine uronic acid, and an unidentified amino sugar. Ruthenium red binding to PS-A2 was stoichiometric: 1 molecule of dye bound per 2.0 carboxyl groups. PMID- 16347547 TI - Establishment of vesicular-arbuscular mycorrhizal fungi and other microorganisms on a beach replenishment site in Florida. AB - Beach replenishment is a widely used method of controlling coastal erosion. To reduce erosional losses from wind, beach grasses are often planted on the replenishment sands. However, there is little information on the microbial populations in this material that may affect plant establishment and growth. The objectives of this research were to document changes in the populations of vesicular-arbuscular mycorrhizal (VAM) fungi and other soil microorganisms in replenishment materials and to determine whether roots of transplanted beach grasses become colonized by beneficial microbes. The study was conducted over a 2 year period on a replenishment project in northeastern Florida. Three sampling locations were established at 1-km intervals along the beach. Each location consisted of three plots: an established dune, replenishment sand planted with Uniola paniculata and Panicum sp., and replenishment sand left unplanted. Fungal and bacterial populations increased rapidly in the rhizosphere of beach grasses in the planted plots. However, no bacteria were recovered that could fix significant amounts of N(2). The VAM fungi established slowly on the transplanted grasses. Even after two growing seasons, levels of root colonization and sporulation were significantly below those found in the established dune. There was a shift in the dominant VAM fungi found in the planted zone with respect to those in the established dunes. The most abundant species recovered from the established dunes were Glomus deserticola, followed by Acaulospora scrobiculata and Scutellospora weresubiae. The VAM fungi that colonized the planted zone most rapidly were Glomus globiferum, followed by G. deserticola and Glomus aggregatum. PMID- 16347546 TI - Effect of 5-fluoro-2'-deoxyuridine on [h]thymidine incorporation by bacterioplankton in the waters of southwest Florida. AB - The effect of 5-fluoro-2'-deoxyuridine (FdUrd) on [methyl-H] thymidine incorporation by bacterioplankton populations in subtropical freshwater, estuarine, and oceanic environments was examined. In estuarine waters, intracellular isotope dilution was inhibited by FdUrd, which enabled us to estimate both intracellular and extracellular isotope dilution. In 2 of 10 cases, extracellular isotope dilution was significant. At low concentrations of [methyl H]thymidine or [6-H]thymidine, FdUrd completely inhibited incorporation of radioactivity into protein and RNA. At high concentrations of [H]thymidine, however, FdUrd had little effect on labeling patterns. The dihydrofolate reductase inhibitors amethopterin and trimethoprim had no effect on macromolecular labeling patterns. These results suggest that thymidylate synthase is not involved in nonspecific labeling and that FdUrd inhibits nonspecific labeling by blocking some other enzyme involved in thymidine catabolism. In oligotrophic oceanic and freshwater samples, FdUrd did not inhibit intracellular isotope dilution or [H]thymidine labeling of protein and RNA, but caused some inhibition of [H]thymidine incorporation into DNA. The ability of FdUrd to inhibit nonspecific macromolecular labeling during [H]thymidine incorporation was significantly correlated (r = 0.84) with total thymidine incorporation (in picomoles per liter per hour). The results are discussed in terms of applications of FdUrd to routine bacterial production measurements and the general assumptions of [H]thymidine incorporation. PMID- 16347548 TI - Production of vesicular-arbuscular mycorrhizal fungus inoculum in aeroponic culture. AB - Bahia grass (Paspalum notatum) and industrial sweet potato (Ipomoea batatas) colonized by Glomus deserticola, G. etunicatum, and G. intraradices were grown in aeroponic cultures. After 12 to 14 weeks, all roots were colonized by the inoculated vesicular-arbuscular mycorrhizal fungi. Abundant vesicles and arbuscules formed in the roots, and profuse sporulation was detected intra-and extraradically. Within each fungal species, industrial sweet potato contained significantly more roots and spores per plant than bahia grass did, although the percent root colonization was similar for both hosts. Mean percent root colonization and sporulation per centimeter of colonized root generally increased with time, although with some treatments colonization declined by week 14. Spore production ranged from 4 spores per cm of colonized root for G. etunicatum to 51 spores per cm for G. intraradices. Infectivity trials with root inocula resulted in a mean of 38, 45, and 28% of bahia grass roots colonized by G. deserticola, G. etunicatum, and G. intraradices, respectively. The germination rate of G. etunicatum spores produced in soil was significantly higher than that produced in aeroponic cultures (64% versus 46%) after a 2-week incubation at 28 degrees C. However, infectivity studies comparing G. etunicatum spores from soil and aeroponic culture indicated no biological differences between the spore sources. Aeroponically produced G. deserticola and G. etunicatum inocula retained their infectivity after cold storage (4 degrees C) in either sterile water or moist vermiculite for at least 4 and 9 months, respectively. PMID- 16347549 TI - Transposon Tn5-Generated Bradyrhizobium japonicum Mutants Unable To Grow Chemoautotrophically with H(2). AB - Twelve Tn5-induced mutants of Bradyrhizobium japonicum unable to grow chemoautotrophically with CO(2) and H(2) (Aut) were isolated. Five Aut mutants lacked hydrogen uptake activity (Hup). The other seven Aut mutants possessed wild type levels of hydrogen uptake activity (Hup), both in free-living culture and symbiotically. Three of the Hup mutants lacked hydrogenase activity both in free living culture and as nodule bacteroids. The other two mutants were Hup only in free-living culture. The latter two mutants appeared to be hypersensitive to repression by oxygen, since Hup activity could be derepressed under 0.4% O(2). All five Hup mutants expressed both ex planta and symbiotic nitrogenase activities. Two of the seven Aut Hup mutants expressed no free-living nitrogenase activity, but they did express it symbiotically. These two strains, plus one other Aut Hup mutant, had CO(2) fixation activities 20 to 32% of the wild-type level. The cosmid pSH22, which was shown previously to contain hydrogenase related genes of B. japonicum, was conjugated into each Aut mutant. The Aut Hup mutants that were Hup both in free-living culture and symbiotically were complemented by the cosmid. None of the other mutants was complemented by pSH22. Individual subcloned fragments of pSH22 were used to complement two of the Hup mutants. PMID- 16347550 TI - Molecular Studies on the Role of a Root Surface Agglutinin in Adherence and Colonization by Pseudomonas putida. AB - Pseudomonas putida aggressively colonizes root surfaces and is agglutinated by a root surface glycoprotein. Mutants of P. putida derived chemically or by Tn5 insertion demonstrated enhanced or decreased agglutinability. Two nonagglutinable Tn5 mutants (Agg) and two mutants with enhanced agglutinability (Agg) possessed Tn5 in unique restriction sites. Agg mutants colonized root surfaces of seedlings grown from inoculated seeds, but at levels lower than those observed with the Agg parent. In short-term binding studies, Agg cells adhered at levels that were 20- to 30-fold less than those for Agg parental cells. These data suggest that the agglutination interaction plays a role in the attachment of P. putida to root surfaces. PMID- 16347551 TI - Oxidative Pathway from Squalene to Geranylacetone in Arthrobacter sp. Strain Y 11. AB - The reaction pathway from squalene to trans-geranylacetone in Arthrobacter sp. strain Y-11 was studied. The enzyme or enzymes catalyzing squalene degradation were found to be membrane bound. Stoichiometric analysis of a cell-free system revealed that the ratio of squalene to trans-geranylacetone changed from 1:2 to 1:1 as the reaction proceeded, indicating two steps in geranylacetone formation. The initial step was found to be oxygenase catalyzed, from the absolute requirement for molecular oxygen in geranylacetone formation and the incorporation of O into geranylacetone under O(2) atmosphere. By using [H]squalene as the substrate, we detected an intermediate in the pathway and identified it as 5,9,13-trimethyltetradeca-4,8,12-trienoic acid by mass spectrometry, infrared spectrometry, nuclear magnetic resonance spectrometry, and chemical synthesis. We deduced that squalene was first oxidatively cleaved to geranylacetone and the intermediate, and that the intermediate was further metabolized to geranylacetone. We also synthesized some of the presumptive metabolites, such as 4,8,12-trimethyltrideca-4,8,12-trien-2-one, and confirmed that they served as active precursors for geranylacetone formation. Based on these lines of evidence, we present here the pathway from squalene to trans geranylacetone in Arthrobacter sp. strain Y-11. PMID- 16347552 TI - Effect of Phosphate on the Corrosion of Carbon Steel and on the Composition of Corrosion Products in Two-Stage Continuous Cultures of Desulfovibrio desulfuricans. AB - A field isolate of Desulfovibrio desulfuricans was grown in defined medium in a two-stage continuous culture apparatus with different concentrations of phosphate in the feed medium. The first state (V1) was operated as a conventional chemostat (D = 0.045 h) that was limited in energy source (lactate) or phosphate. The second stage (V2) received effluent from V1 but no additional nutrients, and contained a healthy population of transiently starved or resting cells. An increase in the concentration of phosphate in the medium fed to V1 resulted in increased corrosion rates of carbon steel in both V1 and V2. Despite the more rapid corrosion observed in growing cultures relative to that in resting cultures, corrosion products that were isolated under strictly anaerobic conditions from the two culture modes had similar bulk compositions which varied with the phosphate content of the medium. Crystalline mackinawite (Fe(9)S(8)), vivianite [Fe(3)(PO(4))(2) . 8H(2)O], and goethite [FeO(OH)] were detected in amounts which varied with the culture conditions. Chemical analyses indicated that the S in the corrosion product was almost exclusively in the form of sulfides, while the P was present both as phosphate and as unidentified components, possibly reduced P species. Some differential localization of S and P was observed in intact corrosion products. Cells from lactate-limited, but not from phosphate-limited, cultures contained intracellular granules that were enriched in P and Fe. The results are discussed in terms of several proposed mechanisms of microbiologically influenced corrosion. PMID- 16347553 TI - Expression of Different Levels of Ethanologenic Enzymes from Zymomonas mobilis in Recombinant Strains of Escherichia coli. AB - The expression of Zymomonas mobilis genes encoding pyruvate decarboxylase and alcohol dehydrogenase II in Escherichia coli converted this organism from the production of organic acids to the production of ethanol. Ethanol was produced during both anaerobic and aerobic growth. The extent to which these ethanologenic enzymes were expressed correlated with the extent of ethanol production. The replacement of organic acids with ethanol as a metabolic product during aerobic and anaerobic growth resulted in dramatic increases in final cell density, indicating that these acids (and the associated decline in pH) are more damaging than the production of ethanol. Of the plasmids examined, the best plasmid for growth and ethanol production expressed pyruvate decarboxylase and alcohol dehydrogenase II at levels of 6.5 and 2.5 IU/mg of total cell protein, respectively. PMID- 16347554 TI - Isolation of a DNA Probe for Lactobacillus curvatus. AB - A genomic library of Lactobacillus curvatus DSM 20019 was constructed in bacteriophage lambda gt11. A 1.2-kilobase DNA probe specific for L. curvatus was isolated from this library. When this probe was hybridized to DNA from Lactobacillus isolates from different sources classified by conventional techniques, differing degrees of hybridization were obtained. This could imply that these isolates may have been incorrectly classified. PMID- 16347555 TI - Induction of Symbiotically Defective Auxotrophic Mutants of Rhizobium fredii HH303 by Transposon Mutagenesis. AB - Symbiotically defective auxotrophic mutants were isolated by transposon Tn5 mutagenesis of Rhizobium fredii HH303, a fast-growing microsymbiont of North American commercial soybean cultivars such as Glycine max cv. Williams. Three different Tn5-carrying suicide vectors, pBLK1-2, pSUP1011, and pGS9, were used for mutagenesis with transposition frequencies of 4 x 10, 3 x 10, and 1 x 10, respectively, while the frequency of background mutation resistant to 500 mug of kanamycin per ml was 1 x 10. From 2,600 Tn5-induced mutants, 14 auxotrophic mutants were isolated and classified in seven groups including adenosine (four), aspartate (two), cysteine or methionine (two), isoleucine and valine (two), nicotinic acid (one), pantothenic acid (one), and uracil (two). All the auxotrophs induced nodulation on soybean, but the symbiotic effectiveness of each mutant was different. Three auxotrophs (two cysteine or methionine and one pantothenic acid) formed effective nodules similar to those of the wild type. Three auxotrophs (one nicotinic acid and two aspartate) produced mature nodules like those of the wild type, but the nodules lacked the characteristic pink color inside and were unable to fix nitrogen. Four auxotrophs (two adenosine and two uracil) induced pseudonodules unable to fix nitrogen. The other four auxotrophs repeatedly induced both effective and ineffective nodules, but bacteroids isolated from the effective nodules were prototrophic revertants. The symbiotic phenotype and the degree of effectiveness of the auxotrophic mutants varied with the type of mutation. PMID- 16347556 TI - Prediction of substrate removal rates of attached microorganisms and of relative contributions of attached and suspended communities at field sites. AB - A mathematical model composed of a direct proportionality relationship between bulk water velocities and field-determined second-order microbial transformation rate coefficients, and the relative rate coefficient of a benchmark chemical, was developed for estimating the substrate removal rates of rapidly degraded chemicals by attached organisms in shallow (<1 m deep) aquatic ecosystems. Data from 31 field experiments involving the addition of 2,4-dichlorophenoxyacetic acid methyl ester (2,4-DME) in nine field areas were used to determine a field derived second-order rate coefficient for microbial transformation of the ester. By using 2,4-DME as a benchmark chemical, the model was used to predict microbial transformation rates of the butoxyethyl ester of 2,4-dichlorophenoxyacetic acid (2,4-DBE) at five other field sites. The predicted half-lives of 2,4-DBE varied 1,500-fold and were within about a threefold range or less of the measured half lives. Under conditions of mass transport limitation, the contributions of attached microorganisms relative to total microbial activities at various field sites were related to the ratio of water velocity, U, and depth, D, showing that historical definitions of ecosystems according to flow and depth characteristics are also valid for describing the process-related structure of ecosystems. An equation was developed for predicting the relative contributions of attached and suspended communities with values of U and D for lotic and lentic ecosystems. On the basis of this equation, attached microorganisms were expected to be insignificant in deep lentic ecosystems and suspended microorganisms were expected to be insignificant in shallow lotic systems for the same process carried out by both populations. Neglecting epiphytic microorganisms, both suspended and attached organisms were expected to be significant in wetlands. PMID- 16347557 TI - Reductive carboxylation of propionate to butyrate in methanogenic ecosystems. AB - During the batch degradation of sodium propionate by the anaerobic sludge from an industrial digestor, we observed a significant amount of butyrate formation. Varying the initial propionate concentrations did not alter the ratio of maximal butyrate accumulation to initial propionate concentration within a large range. By measuring the decrease in the radioactivity of [1-C]butyrate during propionate degradation, we estimated that about 20% of the propionate was converted to butyrate. Labeled butyrate was formed from [1-C]propionate with the same specific radioactivity, suggesting a possible direct pathway from propionate to butyrate. We confirmed this hypothesis by nuclear magnetic resonance studies with [C]propionate. The results showed that [1-C]-, [2-C]-, and [3-C]propionate were converted to [2-C]-, [3-C]-, and [4-C]butyrate, respectively, demonstrating the direct carboxylation on the carboxyl group of propionate without randomization of the other two carbons. In addition, we observed an exchange reaction between C-2 and C-3 of the propionate, indicating that acetogensis may proceed through a randomizing pathway. The physiological significance and importance of various metabolic pathways involved in propionate degradation are discussed, and an unusual pathway of butyrate synthesis is proposed. PMID- 16347558 TI - Relationship of Intracellular Coenzyme F(420) Content to Growth and Metabolic Activity of Methanobacterium bryantii and Methanosarcina barkeri. AB - The use of F(420) as a parameter for growth or metabolic activity of methanogenic bacteria was investigated. Two representative species of methanogens were grown in batch culture: Methanobacterium bryantii (strain M.o.H.G.) on H(2) and CO(2), and Methanosarcina barkeri (strain Fusaro) on methanol or acetate. The total intracellular content of coenzyme F(420) was followed by high-resolution fluorescence spectroscopy. F(420) concentration in M. bryantii ranged from 1.84 to 3.65 mumol . g of protein; and in M. barkeri grown with methanol it ranged from 0.84 to 1.54 mumol . g depending on growth conditions. The content of F(420) in M. barkeri was influenced by a factor of 2 depending on the composition of the medium (minimal or complex) and by a factor of 3 to 4 depending on whether methanol or acetate was used as the carbon source. A comparison of F(420) content with protein, cell dry weight, optical density, and specific methane production rate showed that the intracellular content of F(420) approximately followed the increase in biomass in both strains. In contrast, no correlation was found between specific methane production rate and intracellular F(420) content. However, qCH(4)(F(420)), calculated by dividing the methane production rate by the coenzyme F(420) concentration, almost paralleled qCH(4)(protein). These results suggest that F(420) may be used as a specific parameter for estimating the biomass, but not the metabolic activity, of methanogens; hence qCH(4)(F(420)) determined in mixed populations with complex carbon substrates must be considered as measure of the actual methanogenic activity and not as a measure of potential activity. PMID- 16347559 TI - Purification and Properties of Primary and Secondary Alcohol Dehydrogenases from Thermoanaerobacter ethanolicus. AB - Thermoanaerobacter ethanolicus (ATCC 31550) has primary and secondary alcohol dehydrogenases. The two enzymes were purified to homogeneity as judged from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration. The apparent M(r)s of the primary and secondary alcohol dehydrogenases are 184,000 and 172,000, respectively. Both enzymes have high thermostability. They are tetrameric with apparently identical subunits and contain from 3.2 to 5.5 atoms of Zn per subunit. The two dehydrogenases are NADP dependent and reversibly convert ethanol and 1-propanol to the respective aldehydes. The V(m) values with ethanol as a substrate are 45.6 mumol/min per mg for the primary alcohol dehydrogenase and 13 mumol/min per mg for the secondary alcohol dehydrogenase at pH 8.9 and 60 degrees C. The primary enzyme oxidizes primary alcohols, including up to heptanol, at rates similar to that of ethanol. It is inactive with secondary alcohols. The secondary enzyme is inactive with 1-pentanol or longer chain alcohols. Its best substrate is 2-propanol, which is oxidized 15 times faster than ethanol. The secondary alcohol dehydrogenase is formed early during the growth cycle. It is stimulated by pyruvate and has a low K(m) for acetaldehyde (44.8 mM) in comparison to that of the primary alcohol dehydrogenase (210 mM). The latter enzyme is formed late in the growth cycle. It is postulated that the secondary alcohol dehydrogenase is largely responsible for the formation of ethanol in fermentations of carbohydrates by T. ethanolicus. PMID- 16347560 TI - Influence of Veratryl Alcohol and Hydrogen Peroxide on Ligninase Activity and Ligninase Production by Phanerochaete chrysosporium. AB - Veratryl alcohol, added as a supplement to cultures of Phanerochaete chrysosporium, enhanced ligninase activity through protection of the ligninase against inactivation by hydrogen peroxide produced by this fungus in cultures. In the presence of veratryl alcohol, the loss of ligninase activity observed in non protein-synthesizing cultures (cycloheximide-treated) equaled the extracellular protein turnover. When cultures were not supplemented with veratryl alcohol, inactivation of ligninase by hydrogen peroxide added to protein turnover, resulting in a more rapid loss of ligninase activity. Although all ligninase isoenzymes are sensitive to inactivation by hydrogen peroxide, only the isoenzyme of the highest specific activity (80.6 nkat . mg of protein; M(r), 41,800; pI, 3.96) was found to be protected by veratryl alcohol. The concentration of veratryl alcohol necessary for full protection of ligninase activity varied according to the concentration of hydrogen peroxide present in the medium, which depended on the nature of the carbon source (glucose or glycerol). It is proposed that the nature of the carbon source influences the overall ligninase activity not only directly, by affecting the rate and the type of synthesized ligninase, but also by affecting the rate of hydrogen peroxide production, bringing about different rates of inactivation. PMID- 16347561 TI - Effects of Mannose on the Growth of N(2)-Fixing Azotobacter vinelandii. AB - Mannose is not a suitable substrate for N(2)-fixing Azotobacter vinelandii. However, when H(2) gas is provided, A. vinelandii can grow mixotrophically with H(2) as the energy source and mannose as the carbon source (T.-Y. Wong and R. J. Maier, J. Bacteriol. 163:528-533, 1985). In this report, seven sugars were used to determine whether A. vinelandii could derive energy from these sugars for mannose utilization. Supplementation of fructose- or galactose-limited medium with mannose did not influence the biomass produced by N(2)-fixing A. vinelandii. The presence of mannose in glucose- or maltose-limited cultures increased cell yield slightly. The addition of mannose decreased the total biomass in the melibiose-limited culture slightly. Mannose was a potent inhibitor of growth when sucrose or turanose was used as the primary sugar. The inhibitory effect of mannose on utilization of sucrose and turanose seems to be related to the energy requirement of the N(2)-fixing processes. PMID- 16347562 TI - Expression and Secretion of a Cellulomonas fimi Exoglucanase in Saccharomyces cerevisiae. AB - We used the yeast MEL1 gene for secreted alpha-galactosidase to construct cartridges for the regulated expression of foreign proteins from Saccharomyces cerevisiae. The gene for a Cellulomonas fimi beta-1,4-exoglucanase was inserted into one cartridge to create a fusion of the alpha-galactosidase signal peptide to the exoglucanase. Yeast transformed with plasmids containing this construction produced active extracellular exoglucanase when grown under conditions appropriate to MEL1 promoter function. The cells also produced active intracellular enzyme. The secreted exoglucanase was N-glycosylated and was produced continuously during culture growth. It hydrolyzed xylan, carboxymethyl cellulose, 4-methylumbelliferyl-beta-d-cellobiose, and p-nitrophenyl-beta-d cellobiose. A comparison of the recombinant S. cerevisiae enzyme with the native C. fimi enzyme showed the yeast version to have an identical K(m) and pH optimum but to be more thermostable. PMID- 16347563 TI - Production of Formaldehyde by Detergent-Treated Cells of a Methanol Yeast, Candida boidinii S2 Mutant Strain AOU-1. AB - Treatment of cells of a methanol yeast, Candida boidinii, with the cationic detergent cetyldimethylbenzyl-ammonium chloride (Cation M2) improved the production of formaldehyde. Formaldehyde production was improved twofold with respect to the initial amount of formaldehyde and 1.61-fold with respect to the final amount of formaldehyde after a 12-h reaction under optimized detergent treatment conditions. The treatment caused formaldehyde and formate dehydrogenases to leak out of the cells more rapidly than catalase, but there was no leakage of alcohol oxidase. The improvement in formaldehyde production was considered to be due to the increased permeability of yeast cell membranes and to lower activities of formaldehyde and formate dehydrogenases in Cation M2-treated cells than in intact cells. Changes in the ultrastructure of the cells were observed upon Cation M2 treatment. Several developed peroxisomes were observed in intact cells. After Cation M2 treatment, the cells were obviously damaged, and several peroxisomes seemed to have fused with each other. PMID- 16347564 TI - Expression in Escherichia coli of the Cellulomonas fimi Structural Gene for Endoglucanase B. AB - Endoglucanase B (EB) of Cellulomonas fimi has an M(r) of 110,000 when it is produced in Escherichia coli. The level of expression of the cenB gene (encoding EB) was significantly increased by replacing its normal transcriptional and translational regulatory signals with those of the E. coli lac operon. EB was purified to homogeneity from the periplasmic fraction of E. coli in one step by affinity chromatography on microcrystalline cellulose (Avicel). Alignment of the NH(2)-terminal amino acid sequence with the partial nucleotide sequence of a fragment of C. fimi DNA showed that EB is preceded by a putative signal polypeptide of 33 amino acids. The signal peptide functions and is processed correctly in E. coli, even when its first 15 amino acids are replaced by the first 7 amino acids of beta-galactosidase. The intact EB polypeptide is not required for enzymatic activity. Active polypeptides with M(r)s of 95,000 and 82,000 also appear in E. coli, and a deletion mutant of cenB encodes an active polypeptide with an M(r) of 72,000. PMID- 16347565 TI - Methylation of halogenated phenols and thiophenols by cell extracts of gram positive and gram-negative bacteria. AB - O-methylation of 2,6-dibromophenol was studied in cell extracts prepared from Rhodococcus sp. strain 1395. O-methylation activity was enhanced by the addition of S-adenosyl-l-methionine but was not affected by the addition of 5 methyltetrahydrofolate nor by up to 10 mM MgCl(2) or EDTA. By using 2,6 dibromophenol, 4,5,6-trichloroguaiacol, and pentachlorothiophenol as the substrates, O-methylation activity was also demonstrated in extracts from two other Rhodococcus sp. strains, an Acinetobacter sp. strain, and a Pseudomonas sp. strain. A diverse range of chloro- and bromophenols, chlorothiophenols, chloro- and bromoguaiacols, and chloro- and bromocatechols were assayed as the substrates by using extracts prepared from strain 1395; all of the compounds were methylated to the corresponding anisoles, veratroles, or guaiacols, which have been identified previously from experiments using whole cells. The specific activity of the enzyme towards the thiophenols was significantly higher than it was towards all the other substrates-high activity was found with pentafluorothiophenol, although the activity with pentafluorophenol was undetectable with the incubation times used. For the chlorophenols, the position of the substituents was of cardinal importance. The enzyme had higher activity towards the halogenated catechols than towards the corresponding guaiacols, and selective O-methylation of the 3,4,5-trihalogenocatechols yielded predominantly the 3,4,5-trihalogenoguaiacols. As in experiments with whole cells, neither 2,4 dinitrophenol, hexachlorophene, nor 5-chloro- or 5-bromovanillin was O methylated. The results showed conclusively that the methylation reactions were enzymatic and confirmed the conclusion from extensive studies using whole cells that methylation of halogenated phenols may be a significant alternative to biodegradation. PMID- 16347566 TI - Preparation and Purification of Xylitol-5-Phosphate from a Cell Extract of Lactobacillus casei Cl-16. AB - A simple procedure which yields pure xylitol-5-phosphate is described. A cell extract of Lactobacillus casei Cl-16 from a 6-liter culture was used to synthesize up to 70 mg of xylitol-5-phosphate overnight from xylitol and phosphoenolpyruvate via a xylitol phosphoenolpyruvate:phosphotransferase system with a 53% yield. Centrifugation, filtration, precipitation as a barium salt, and ion-exchange batch chromatography permitted recovery of nearly 90% of the phosphorylated product synthesized. Thin-layer chromatography and enzymatic analysis indicated a purity level of more than 99%. The method was used to synthesize [U-C]xylitol-5-phosphate, and it is suitable for the synthesis of many other nonmetabolizable sugar phosphates. PMID- 16347567 TI - A Positive Strain Identification Method for Rhizobium meliloti. AB - About 80% of Rhizobium meliloti strains contain 1 to 11 copies of insertion sequence ISRm1 in their genomes (R. Wheatcroft and R. J. Watson, J. Gen. Microbiol. 134:113-121, 1988). Hybridization to separated genomic DNA fragments with an ISRm1-specific probe produces patterns of hybridization bands which are distinctive for each strain. These patterns can be compared between strains to prove or disprove common identity. In most cases relatedness can be inferred despite phenotypic differences or minor genomic alterations. PMID- 16347568 TI - Preparation of a Cell-Free, Isobutene-Forming System from Rhodotorula minuta. AB - The cell-free isobutene-forming system of Rhodotorula minuta IFO 1102 was prepared. This system consisted of isovaleric acid, NADPH, and a cell extract. The maximal activity obtained by using the described cell-free system was 17 nl/mg of protein per h. It appears that this cell-free isobutene-forming system operates in living cells of R. minuta. PMID- 16347569 TI - Biosynthesis of Territrems by Aspergillus terreus. AB - Different radioactive precursors were added to 8-day potato-dextrose liquid cultures of Aspergillus terreus 23-1. Territrems were isolated from chloroform extracts of the cultures at day 14 and purified by thin-layer chromatography and high-pressure liquid chromatography. The territrem B obtained was treated with alkaline hydrogen peroxide, and 3, 4, 5-trimethoxy benzoic acid was isolated from an ethyl acetate extract of the reaction mixture and purified by thin-layer chromatography and high-pressure liquid chromatography. By comparison of the specific radioactivities of territrem B and its cleaved aromatic product (disintegrations per minute per micromole of compound), it was demonstrated that the radioactivity of territrem B was located mainly on its aromatic moiety when [U-C]shikimate, l-[methyl-C]methionine, and l-[methyl-H]methionine were precursors; however, the radioactivity of territrem B was located mainly on its nonaromatic moiety when [2-C]mevalonate was the precursor. Mevinolin, a specific inhibitor of beta-hydroxyl beta-methyl glutaryl coenzyme A reductase, was shown to inhibit production of territrems by A. terreus 23-1. When [U-C]acetate was used as a precursor, mevinolin inhibited the incorporation of radioactive carbon into territrem but mevinolin did not inhibit incorporation of radioactive carbon from [2-C]mevalonate into territrem. PMID- 16347570 TI - Changes in Endoglucanase Patterns during Growth of Thermomonospora curvata on Cellulose. AB - The endoglucanases of the thermophilic actinomycete Thermomonospora curvata were characterized. Early-exponential-phase culture fluid contained at least three endoglucanases, with molecular weights of 23,000, 46,000, and 146,000 and K(m) values of 1.54, 3.60, and 1.32% carboxymethyl cellulose, respectively. The stationary-phase pattern was altered to include three enzymes with molecular weights of 52,000, 114,000, and 106,000, with respective K(m) values of 1.77, 8.30, and 1.91%. PMID- 16347571 TI - Resistance of Thermus spp. to Potassium Tellurite. AB - Two members of the genus Thermus were examined for their resistance to toxic inorganic compounds. They both proved to be fairly resistant to tellurite and selenite and to many other heavy metal salts. Cell extracts of Thermus thermophilus HB8 and of T. flavus AT-62 catalyze the reduction of K(2)TeO(3) in a reaction which is dependent on NADH oxidation. PMID- 16347572 TI - Aggregated and free bacteria as food sources for heterotrophic microflagellates. AB - Laboratory evidence for the selective grazing of aggregated bacteria by a Bodo sp. and free bacteria by a Paraphysomonas sp. is presented. The relevance of different microflagellate feeding mechanisms with respect to prey distribution in the natural environment is discussed. PMID- 16347573 TI - Enumeration of thermophilic heterotrophs in geothermally heated soils from mount erebus, ross island, antarctica. AB - Soil samples with temperatures up to 64 degrees C were collected from Mount Erebus, an active volcano located on Ross Island, Antarctica. Acridine orange direct counts and most probable number counts of soil samples stored at 4 degrees C for 2 months showed a wide variation in the number of thermophilic microorganisms in different soils. Organisms similar to Clostridium thermohydrosulfuricum, Bacillus schlegelii, and Bacillus acidocaldarius, as well as neutrophilic Bacillus strains, were isolated. PMID- 16347574 TI - Chemotaxis by Pseudomonas syringae pv. tomato. AB - Optimal laboratory conditions for studying chemotaxis by Pseudomonas syringae pv. tomato were determined by using the Adler capillary tube assay. Although they are not an absolute requirement for chemotaxis, the presence of 0.1 mM EDTA and 1 mM MgCl(2) in the chemotaxis buffer (10 mM potassium phosphate [pH 7.2]) significantly enhanced the response to attractant. The addition of mannitol as an energy source had little effect. The optimal temperature for chemotaxis was 23 degrees C, which is 5 degrees C below the optimal growth temperature for this pathogen. The best response occurred when the bacteria were exposed to attractant for 60 min at a concentration of approximately 5 x 10 CFU/ml. P. syringae pv. tomato was strongly attracted to citric and malic acids, which are the predominant organic acids in tomato fruit. With the exception of asparagine, the major amino acids of tomatoes were weak to moderate attractants. Glucose and fructose, which account for approximately 47% of tomato dry matter, also elicited poor responses. In assays with tomato intercellular fluid and leaf surface water, the bacterial speck pathogen could not chemotactically distinguish between a resistant and a susceptible cultivar of tomato. PMID- 16347575 TI - Expression of Solvent-Forming Enzymes and Onset of Solvent Production in Batch Cultures of Clostridium beijerinckii ("Clostridium butylicum"). AB - Clostridium beijerinckii ("Clostridium butylicum") NRRL B592 and NRRL B593 were grown in batch cultures without pH control. The use of more sensitive and accurate procedures for the determination of solvents in cultures led to the recognition of the onset of solvent production about 2 h earlier than the previously assigned point and at a higher culture pH for both strains. Reliable assays for solvent-forming enzyme activities in cell extracts have also been developed. The results showed that activities of solvent-forming enzymes in strain NRRL B592 started to increase about 1 h before the measured onset of solvent production and that the increase in activities of solvent-forming enzymes was not simultaneous. The degree of increase of these enzyme activities for both strains ranged from 2- to 165-fold, with acetoacetate decarboxylase and butanol isopropanol dehydrogenase showing the largest activity increases. However, the pattern of increase of enzyme activities differed significantly in the two strains of C. beijerinckii. When an increase in solvent-forming enzyme activities was first detected in strain NRRL B592, the culture pH was at 5.7 and the concentrations of total acetic and butyric acids were 5.2 and 3.6 mM, respectively. For strain NRRL B593, the corresponding pH was 5.5. Thus, the culture conditions immediately preceding the expression of solvent-forming enzyme activities differed significantly from those that have been correlated with the production of solvents at later stages of growth. PMID- 16347576 TI - A Simple and Rapid Method for Genetic Transformation of Lactic Streptococci by Electroporation. AB - An electroporation procedure for the plasmid-mediated genetic transformation of intact cells of Streptococcus cremoris and Streptococcus lactis was performed. Ten different strains were transformed. The method was simple and rapid and yielded transformant colonies in 14 to 24 h. The method was optimized for S. lactis LM0230, and transformation frequencies of between 1 x 10 and 5 x 10 transformants per mug of purified plasmid (pMU1328) were achieved routinely. The optimized procedure involved lysozyme treatment of cells. Transformation of LM0230 occurred at comparable frequencies with pLS1 (4.4 kilobase pair [kbp]), pMU1328 (7.4 kbp), and pAMbeta1 (26.5 kbp). Plasmid DNA isolated from transformants had not undergone detectable deletions or rearrangements. Transformation was possible with plasmid DNA which was religated after restriction endonuclease digestion. Phage DNA-dependent transfection of S. lactis LM0230 and S. lactis C6 was also achieved. PMID- 16347577 TI - Primary and Bacterial Secondary Production in a Southwestern Reservoir. AB - Rates of primary and bacterial secondary production in Lake Arlington, Texas, were determined. The lake is a warm (annual temperature range, 7 to 32 degrees C), shallow, monomictic reservoir with limited macrophyte development in the littoral zone. Samples were collected from six depths within the photic zone from a site located over the deepest portion of the lake. Primary production and bacterial production were calculated from NaHCO(3) and [methyl-H]thymidine incorporation, respectively. Peak instantaneous production ranged between 14.8 and 220.5 mug of C liter h. There were two distinct periods of high rates of production. From May through July, production near the metalimnion exceeded 100 mug of C liter h. During holomixis, production throughout the water column was in excess of 100 mug of C liter h and above 150 mug of C liter h near the surface. Annual areal primary production was 588 g of C m. Bacterial production was markedly seasonal. Growth rates during late fall through spring were typically around 0.002 h, and production rates were typically 5 mug of C liter h. Growth rates were higher during warmer parts of the year and reached 0.03 h by August. The maximum instantaneous rate of bacterial production was approximately 45 mug of C liter h. Annual areal bacterial production was 125 g of C m. Temporal and spatial distributions of bacterial numbers and activities coincided with temporal and spatial distributions of primary production. Areal primary and bacterial secondary production were highly correlated (r = 0.77, n = 15, P < 0.002). PMID- 16347578 TI - Adsorption of Lithocholic Acid to Fusarium equiseti M41 as an Essential Process in Its Conversion to Ursodeoxycholic Acid. AB - Fusarium equiseti M41 converts lithocholic acid to ursodeoxycholic acid. Adsorption of lithocholic acid particles to mycelia of F. equiseti M41 is essential in the conversion of lithocholic acid to ursodeoxycholic acid. Production of ursodeoxycholic acid was negligible when particles of lithocholic acid were absent. As the concentration of lithocholic acid particles increased, both the amount of mycelium-bound lithocholic acid and the production of ursodeoxycholic acid increased hyperbolically (K(1/2) = 1.9 g/liter and K(m(apparent) ) = 1.9 g/liter. A fluorescent lithocholic acid derivative was used to confirm that insoluble particles of lithocholic acid attached to the surface of the mycelia. The hydrophobic nature of this binding was estimated from the close relationship observed between the hydrophobicity of bile acids and their binding capacity to the mycelia. By repeated washing with 30% dimethyl sulfoxide, two binding modes of lithocholic acid were distinguished, i.e., surface binding (59% of bound lithocholic acid) and tight binding (41% of bound lithocholic acid). From the amount of tightly bound lithocholic acid, the intracellular concentration of lithocholic acid was calculated to be 1,433-fold higher than its saturating concentration in the reaction mixture, thus promoting effective conversion to ursodeoxycholic acid in the mycelia. Several lines of evidence indicated that glycoproteins of the cell wall participated in the binding of lithocholic acid. PMID- 16347579 TI - Cloning and Expression of a Clostridium acetobutylicum Alcohol Dehydrogenase Gene in Escherichia coli. AB - An alcohol dehydrogenase (ADH) gene from Clostridium acetobutylicum was cloned on a recombinant plasmid, pCADH100. Escherichia coli HB101, and an allyl alcohol resistant mutant, HB101-adh1, containing this plasmid were unable to grow aerobically or anaerobically on agar media containing sublethal concentrations of allyl alcohol. E. coli HB101 and HB101-adh1 transformed with the plasmid pCADH100 produced increased levels of ethanol when grown anaerobically under alkaline conditions in the absence of nitrate. Cell extracts from aerobically and anaerobically grown E. coli HB101(pCADH100) and HB101-adhl(pCADH100) cells exhibited increased levels of NADP-dependent ADH activity with either ethanol or butanol as the substrate. The inability of E. coli HB101(pCADH100) to grow in the presence of allyl alcohol correlated with the appearance of an NADP-dependent ADH activity band on nondenaturing polyacrylamide gel electrophoresis with either ethanol or butanol as the substrate. The position of the cloned NADP-dependent ADH activity bands in E. coli HB101(pCADH100) cell extracts with either ethanol or butanol as the substrate coincided with the position of a single NADP dependent ADH activity band in extracts of C. acetobutylicum cells. E. coli HB101(pCADH100) cell extracts prepared from both aerobically and anaerobically grown cells exhibited an additional protein band with an apparent M(r) of approximately 33,000 on sodium dodecyl sulfate-polyacryl-amide gel electrophoresis which was absent in cell extracts of E. coli HB101. A protein band with a similar apparent M(r) was observed in cell extracts of C. acetobutylicum, and in vitro transcription and translation experiments with pCADH100 produced a major protein product with a similar apparent M(r). PMID- 16347580 TI - Effects of Temperature and Relative Humidity on the Growth of and Enzyme Production by Actinomucor taiwanensis during Sufu Pehtze Preparation. AB - The growth of and production of protease, alpha-amylase, alpha-galactosidase, and lipase by Actinomucor taiwanensis in relation to temperature and relative humidity during the preparation of sufu (Chinese cheese) pehtze were investigated. The incubation temperature, humidity, and cultivation time greatly affected the growth of and enzyme production by A. taiwanensis on tofu. It grew best at 97% humidity and 30 degrees C. The highest yields of protease (112 U/g of dry tofu) and lipase (1,448 U/g of dry tofu) were found after 60 h of incubation at 97% humidity and 25 degrees C. On the other hand, the highest yield of alpha amylase (1,949 U/g of dry tofu) was observed after 48 h of incubation at 96 to 97% humidity and 30 degrees C, and the highest amount of alpha-galactosidase (387 U/g of dry tofu) was observed at 35 degrees C and 96% humidity after 60 h of growth. The results suggest that the temperature and humidity should be controlled at 25 to 30 degrees C and around 97%, respectively, during the commercial preparation of sufu pehtze for better growth of and production of enzymes by A. taiwanensis. PMID- 16347581 TI - Direct characterization of methanogens in two high-rate anaerobic biological reactors. AB - The methanogenic flora from two types of turbulent, high-rate reactors was studied by immunologic methods as well as by phase-contrast, fluorescence, and scanning electron microscopy. The reactors were a fluidized sand-bed biofilm ANITRON reactor and an ultrafiltration membrane-associated suspended growth MARS reactor (both trademarks of Air Products and Chemicals, Inc., Allentown, Pa.). Conventional microscopic methods revealed complex mixtures of microbes of a range of sizes and shapes, among which morphotypes resembling Methanothrix spp. and Methanosarcina spp. were noticed. Precise identification of these and other methanogens was accomplished by antigenic fingerprinting with a comprehensive panel of calibrated antibody probes of predefined specificity spectra. The methanogens identified showed morphotypes and antigenic fingerprints indicating their close similarity with the following reference organisms: Methanobacterium formicicum MF and Methanosarcina barkeri W in the ANITRON reactor only; Methanosarcina barkeri R1M3, M. mazei S6, Methanogenium cariaci JR1, and Methanobrevibacter arboriphilus AZ in the MARS reactor only; and Methanobrevibacter smithii ALI and Methanothrix soehngenii Opfikon in both reactors. Species diversity and distribution appeared to be, at least in part, dependent on the degree of turbulence inside the reactor. PMID- 16347582 TI - DNA Probe Method for the Detection of Specific Microorganisms in the Soil Bacterial Community. AB - We developed a protocol which yields purified bacterial DNA from the soil bacterial community. The bacteria were first dispersed and separated from soil particles in the presence of polyvinylpolypyrrolidone, which removes humic acid contaminants by adsorption to this insoluble polymer. The soil bacteria were then collected by centrifugation and lysed by using a comprehensive protocol designed to maximize disruption of the various types of bacteria present. Total bacterial DNA was purified from the cell lysate and remaining soil contaminants by using equilibrium density gradients. The isolated DNA was essentially pure as determined by UV spectral analysis, was at least 48 kilobases long, and was not subject to degradation, which indicated that there was no contaminating nuclease activity. The isolated DNA was readily digested by exogenously added restriction endonucleases and successfully analyzed by slot blot and Southern blot hybridizations. Using single-stranded, P-labeled DNA probes, we could detect and quantitate the presence of a specific microbial population in the natural soil community on the basis of the presence of a DNA sequence unique to that organism. The sensitivity of our methodology was sufficient to detect Bradyrhizobium japonicum at densities as low as 4.3 x 10 cells per g (dry weight) of soil, which corresponds to about 0.2 pg of hybridizable DNA in a 1-mug DNA sample. PMID- 16347583 TI - Seasonal and Diel Variability in Dissolved DNA and in Microbial Biomass and Activity in a Subtropical Estuary. AB - Dissolved DNA and microbial biomass and activity parameters were measured over a 15-month period at three stations along a salinity gradient in Tampa Bay, Fla. Dissolved DNA showed seasonal variation, with minimal values in December and January and maximal values in summer months (July and August). This pattern of seasonal variation followed that of particulate DNA and water temperature and did not correlate with bacterioplankton (direct counts and [H]thymidine incorporation) or phytoplankton (chlorophyll a and CO(2) fixation) biomass and activity. Microautotrophic populations showed maxima in the spring and fall, whereas microheterotrophic activity was greatest in late summer (September). Both autotrophic and heterotrophic microbial activity was greatest at the high estuarine (low salinity) station and lowest at the mouth of the bay (high salinity station), irrespective of season. Dissolved DNA carbon and phosphorus constituted 0.11 +/- 0.05% of the dissolved organic carbon and 6.6 +/- 6.5% of the dissolved organic phosphorus, respectively. Strong diel periodicity was noted in dissolved DNA and in microbial activity in Bayboro Harbor during the dry season. A noon maximum in primary productivity was followed by an 8 p.m. maximum in heterotrophic activity and a midnight maximum in dissolved DNA. This diel periodicity was less pronounced in the wet season, when microbial parameters were strongly influenced by episodic inputs of freshwater. These results suggest that seasonal and diel production of dissolved DNA is driven by primary production, either through direct DNA release by phytoplankton, or more likely, through growth of bacterioplankton on phytoplankton exudates, followed by excretion and lysis. PMID- 16347584 TI - Optimization of Banana Juice Fermentation for the Production of Microbial Oil. AB - Apiotrichum curvatum ATCC 20509 (formerly Candida curvata D), a lipid accumulating yeast, was grown in banana juice. The optimum conditions for biomass production in shake flasks were 30 degrees C growth temperature, efficient aeration, a juice concentration of 25%, and preliminary heat treatment at less than sterilization conditions. Under controlled conditions in a fermentor, 20% banana juice was optimum. High concentrations of yeast extract (0.3%) increased biomass production by 40% but decreased oil production by 30%. A lower yeast extract concentration (0.05%) increased biomass production by 2% and oil production by 25%. The best growth and oil production were observed when asparagine (1.4 g/liter) and mineral salts were added to the banana juice. The addition of minerals seemed to improve the utilization of carbon. Growth inhibition was observed when the fermentor was aerated with pure oxygen, even when additional nutrients were present. A fed-batch process permitted the juice concentration to be increased from 15 to 82%; biomass accumulation was three times higher than in batch fermentations. However, the cellular lipid content was only 30% of dry weight, and chemical oxygen demand reduction was slow and inefficient. PMID- 16347585 TI - Conjugal Transfer in Lactic Streptococci of Plasmid-Encoded Insensitivity to Prolate- and Small Isometric-Headed Bacteriophages. AB - Eight of 40 strains of Streptococcus lactis and S. lactis subsp. diacetylactis were able to conjugally transfer a degree of phage insensitivity to Streptococcus lactis LM0230. Transconjugants from one donor strain, S. lactis subsp. diacetylactis 4942, contained a 106-kilobase (kb) cointegrate plasmid, pAJ1106. The plasmid was conjugative (Tra) and conferred phage insensitivity (Hsp) and lactose-fermenting ability (Lac) in S. lactis and Streptococcus cremoris transconjugants. The phage resistance mechanism was effective against prolate- and small isometric-headed phages at 30 degrees C. In S. lactis transconjugants, the phage resistance mechanism was considerably weakened at elevated temperatures. A series of deletion plasmids was isolated from transconjugants in S. cremoris 4854. Deletion plasmids were pAJ2074 (74 kb), Lac, Hsp, Tra; pAJ3060 (60 kb), Lac, Hsp; and pAJ4013 (13 kb), Lac. These plasmids should facilitate mapping Hsp and tra genes, with the aim of constructing phage-insensitive strains useful to the dairy industry. PMID- 16347586 TI - Microbial Decomposition in Aquatic Environments: Combined Process of Extracellular Enzyme Activity and Substrate Uptake. AB - The aim of this study was to define a model for the coupling between extracellular enzyme activity and substrate uptake by bacterial populations in natural waters. The balance between uptake of leucine and extracellular hydrolytic production of leucine from a peptide model substrate was investigated in a combined fluorescence-radiotracer experiment with [H]leucine as a marker for the leucine pool and l-leucine-4 methyl-7-coumarinylamide (Leu-MCA) as a marker for the pool of dissolved peptide substrates. Results show that at low concentrations of the model substrate the input and uptake processes of leucine are nearly balanced, whereas at high concentrations of the model substrate much more leucine is liberated than taken up. In addition, samples from one polluted and one less polluted station in the Kiel Fjord were investigated for their extracellular enzymatic and uptake properties in an annual cycle. It was found that turnover rates of leucine (T(r), percent per hour) and hydrolysis rates of Leu-MCA (H(r), percent per hour), as well as the quotient T(r)/H(r), reflect the impact of environmental conditions on decomposition processes at both sampling sites. The quotient T(r)/H(r) is interpreted as an indirect measurement of the pool size ratio (polymers/monomers), which may serve as an index of hydrolysis uptake coupling in bacterial utilization of dissolved protein. Calculated on an annual average basis, turnover rates are ca. nine times higher than hydrolysis rates at the polluted station and ca. five times higher at the less polluted station. From the described model, this would mean that the relative fraction of polymers within the total dissolved organic carbon pool (with regard to the substrate combination dissolved protein-leucine) is about twice that at the polluted than at the less polluted station. PMID- 16347587 TI - Dual-Label Radioisotope Method for Simultaneously Measuring Bacterial Production and Metabolism in Natural Waters. AB - Bacterial production and amino acid metabolism in aquatic systems can be estimated by simultaneous incubation of water samples with both tritiated methyl thymidine and C-labeled amino acids. This dual-label method not only saves time, labor, and materials, but also allows determination of these two parameters in the same microbial subcommunity. Both organic carbon incorporation and respiration can be estimated. The results obtained with the dual-label technique are not significantly different from single-radiolabel methods over a wide range of bacterial activity. The method is particularly suitable for large-scale field programs and has been used successfully with eutrophic estuarine samples as well as with oligotrophic oceanic water. In the mesohaline portion of Chesapeake Bay, thymidine incorporation ranged seasonally from 2 to 635 pmol liter h and amino acid turnover rates ranged from 0.01 to 28.4% h. Comparison of thymidine incorporation with amino acid turnover measurements made at a deep, midbay station in 1985 suggested a close coupling between bacterial production and amino acid metabolism during most of the year. However, production-specific amino acid turnover rates increased dramatically in deep bay waters during the spring phytoplankton bloom, indicating transient decoupling of bacterial production from metabolism. Ecological features such as this are readily detectable with the dual label method. PMID- 16347588 TI - Application of the Fluorescent-Antibody Technique for the Detection of Sphaerotilus natans in Activated Sludge. AB - Sphaerotilus natans, one of the most widely reported causes of bulking in activated sludge, can exist both within and outside of a sheath. It can easily be confused with similar activated sludge bacteria and thus can be overlooked when present in low numbers. Fluorescent antiserum was successfully prepared against the nonfilamentous form and was shown to be highly specific, showing no reaction with either pure cultures of similar filamentous bacteria or entirely unrelated organisms. It did, however, show a lack of strain specificity since it reacted with S. natans isolates from the Federal Republic of Germany and the United States and with filamentous bacteria in South African activated sludges. Fluorescent antibody is capable of penetrating the filaments of S. natans to stain the cells individually. The use of fluorescent antiserum in the identification of S. natans filaments obscured by activated sludge flocs and other suspended matter was simple since the cells stained brightly and could be observed through the less dense matter, while the use of other microscope techniques would be hampered by these obstructions. The use of fluorescent antibody will facilitate ecological studies of S. natans in activated sludge and other aqueous environments. PMID- 16347589 TI - Influence of Glycine spp. on Competitiveness of Bradyrhizobium japonicum and Rhizobium fredii. AB - The displacement of indigenous Bradyrhizobium japonicum in soybean nodules with more effective strains offers the possibility of enhanced N(2) fixation in soybean (Glycine max (L.) Merr.). Our objective was to determine whether the wild soybean (G. soja Sieb. & Zucc.) genotype PI 468397 would cause reduced competitiveness of important indigenous B. japonicum strains USDA 31, 76, and 123 and thereby permit nodulation by Rhizobium fredii, the fast-growing microsymbiont of soybean. In an initial experiment, PI 468397 nodulated and fixed moderate amounts of N(2) with USDA 31 and 76 but, despite the formation of nodules, fixed essentially no N(2) with USDA 123. In contrast, PI 468397 formed a highly effective symbiosis with R. fredii strain USDA 193. In two subsequent experiments, Williams soybean and PI 468397 were grown in a pasteurized soil mixture or in soybean rhizobium-free soil and inoculated with both USDA 123 and USDA 193. In each experiment, more than 90% of the nodules of Williams contained USDA 123, while only a maximum of 2% were occupied with USDA 193. In contrast, in the two experiments, 16 and 11%, respectively, of the nodules produced on PI 468397 were occupied by USDA 123, while in both experiments 87% contained USDA 193. Thus, in relation to the cultivar Williams, which is commonly grown and used as a parent in soybean breeding programs in the United States, PI 468397 substantially reduced the competitive ability of B. japonicum strain USDA 123 in relation to R. fredii strain USDA 193. PMID- 16347590 TI - Isolation and Characterization of Marine Caulobacters and Assessment of Their Potential for Genetic Experimentation. AB - A total of 25 marine caulobacters were isolated from littoral marine sources. Several aspects of their physiology and morphology were examined, as well as their suitability for genetic manipulation in laboratory cultivation. Caulobacters were readily isolated from all sources, including samples from areas containing pollution-related organic compounds. All isolates grew best in media containing seawater, but eight strains grew if sea salts were replaced with NaCl alone, three strains grew at 1/10 the normal sea salt concentration, and one isolate grew, albeit poorly, in freshwater medium. Of the marine isolates, 12 strains grew under anaerobic conditions, indicating that some caulobacters are not obligately aerobic bacteria, as they are currently categorized. Although some freshwater caulobacters are able to oxidize manganese, this capability was not found in these marine caulobacters. Of the marine isolates, 10 strains were resistant to mercury chloride concentrations 10- to 20-fold greater than that tolerated by sensitive bacteria. However, a mercury reductase gene comparable with that found in R100-type plasmids was not detected by gene hybridization. With respect to the potential for genetic experimentation, most strains grew rapidly (3- to 4-h generation time at 30 degrees C), producing colonies on solid media in 2 to 3 days. The isolates were sensitive to antibiotics commonly used in recombinant DNA experiments, and spontaneous drug-resistant mutants were selectable. Conjugal transfer of plasmids from Escherichia coli to several marine caulobacters was demonstrated for four broad-host-range plasmid incompatibility groups, by using both self-transmissible plasmids and cloning-oriented plasmids that require a helper plasmid. Conjugal transfer of broad-host-range plasmids between freshwater and marine caulobacters was also demonstrated in both directions. Native plasmids of approximately 100- to 150-kilobase sizes were found in 2 of the 25 marine Caulobacter strains. The native plasmids were present in relatively high copy number and appeared stable in laboratory culture. In short, the marine caulobacters appeared appropriate as candidates for genetic manipulation and the expression of selected genes in the marine environment. PMID- 16347591 TI - Isolation and characterization of methanogenic bacteria from landfills. AB - Methanogenic bacteria were isolated from landfill sites in the United Kingdom. Strains of Methanobacterium formicicum, Methanosarcina barkeri, several different immunotypes of Methanobacterium bryantii, and a coccoid methanogen distinct from the reference immunotypes were identified. PMID- 16347592 TI - Effect of Iron Limitation on "Pseudomonas plantarii" Growth and Tropolone and Protein Production. AB - The addition of iron to an iron-deficient medium markedly enhanced the growth of "Pseudomonas plantarii" and the production of red crystals. However, it markedly reduced the amount of dissolved tropolone, a product of the bacterium and an iron chelator, and the production of an iron-regulated protein (78 kilodaltons). The red crystal was complex, composed of tropolone and iron, the ratio being 3:1. PMID- 16347593 TI - Effect of Naturally Occurring nif Reiterations on Symbiotic Effectiveness in Rhizobium phaseoli. AB - Most naturally occurring strains of Rhizobium phaseoli possess reiteration of the nif genes. Three regions contain nitrogenase structural genes in strain CFN42. Two of these regions (a and b) have copies of nifH, nifD, and nifK, whereas the third region (c) contains only nifH. Strains containing mutations in either nif region a or nif region b had significantly diminished symbiotic effectiveness compared with the wild-type strain on the basis of nodule mass, total nitrogenase activity per plant, nitrogenase specific activity, total nitrogen in the shoot, and percentage of nitrogen. A strain containing mutations in both nif region a and nif region b was totally ineffective. These data indicate that both nif region a and nif region b are needed for full symbiotic effectiveness in R. phaseoli. PMID- 16347594 TI - Enzyme-Linked Immunosorbent Assays for the Specific and Sensitive Quantification of Methanosarcina mazei and Methanobacterium bryantii. AB - Three microtitration plate enzyme-linked immunosorbent assays (ELISAs) have been developed: a competitive ELISA and a two-site (or indirect sandwich) ELISA for Methanosarcina mazei S6 and a two-site ELISA for Methanobacterium bryantii FR-2. The assays were sensitive, with limits of cell protein detection of 3 ng ml, 5 ng ml, and 50 ng ml, respectively, and showed good precision. The M. mazei assays used monoclonal antibodies and were entirely species specific, showing no cross reaction with methanogens of other genera or with other species of the same genus. The Methanobacterium bryantii assay, which used two polyclonal antisera, showed only a slight cross-reaction with one other Methanobacterium species but no cross-reaction with methanogens of other genera. The use of the ELISAs for quantitative analysis of mixed cultures and of sewage sludge samples was investigated. Sludge diluted at 1:10 or more caused no significant interference in any of the three ELISAs. Various cultures of bacteria, methanogens, and nonmethanogens at a protein concentration of 50 mug ml showed no significant interference in the M. mazei competitive assay and the Methanobacterium bryantii two-site assay, although they did cause falsely low results in the M. mazei two site assay. PMID- 16347595 TI - Anaerobic microbial dissolution of transition and heavy metal oxides. AB - Anaerobic microbial dissolution of several crystalline, water-insoluble forms of metal oxides commonly associated with the waste from energy production was investigated. An anaerobic N-fixing Clostridium sp. with an acetic, butyric, and lactic acid fermentation pattern, isolated from coal-cleaning waste, solubilized Fe(2)O(3) and MnO(2) by direct enzymatic reduction; CdO, CuO, PbO, and ZnO were solubilized by indirect action due to the production of metabolites and the lowering of the pH of the growth medium. Extracellular heat-labile components of the cell-free spent medium obtained from cultures without oxide solubilized a significant amount of Fe(2)O(3) (1.7 mumol); however, direct contact with the bacterial cells resulted in the complete dissolution (4.8 mumol) of the oxide. Under identical conditions, the cell-free spent medium solubilized only a small amount of MnO(2) (0.07 mumol), whereas 2.3 mumol of the oxide was solubilized by direct bacterial contact. Reduction of Fe(2)O(3) and MnO(2) by Clostridium sp. proceeds at different rates and, possibly, by different enzymatic systems. Fe(III) and Mn(IV) oxides appear to be used as sinks for excess electrons generated from glucose fermentation, since there is no apparent increase in growth of the bacterium concomitant with the reduction of the oxides. Dialysis bag experiments with Co(2)O(3) indicate that there is a slight dissolution of Co (0.16 mumol) followed by precipitation or biosorption. Although Mn(2)O(3), Ni(2)O(3), and PbO(2) may undergo reductive dissolution from a higher to a lower oxidation state, dissolution by direct or indirect action was not observed. Also, Cr(2)O(3) and NiO were not solubilized by direct or indirect action. Significant amounts of solubilized Cd, Cu, and Pb were immobilized by the bacterial biomass, and the addition of Cu inhibited the growth of the bacterium. PMID- 16347596 TI - Biotransformation of Dehydroabietic, Abietic, and Isopimaric Acids by Mortierella isabellina Immobilized in Polyurethane Foam. AB - Polyurethane foam supports immobilization of Mortierella isabellina ATCC 38063, a zygomycete that hydroxylates and thereby detoxifies resin acids found in certain pulp mill effluents. The fungus becomes entrapped as it threads growing mycelia through the open foam matrix. The tenacity of binding and the amount of biomass bound depend on initial spore numbers and nutrient concentration. Optimal dehydroabietic acid transformation occurs with early-stationary-phase foam-bound mycelia suspended in buffer at pH 6.5 to 8.5 with aeration >0.1 liter liter min and near a temperature maximum of 33 degrees C. Neither a greater number of similarly loaded foam cubes nor a greater surface area for similar amounts of biomass increased transformation rates. Compared to free mycelial cultures, foam presence retards growth and decreases the rate of precursor transformation but does not alter the nature of the metabolites formed from dehydroabietic acid, abietic acid, or isopimaric acid. Diffusional barriers or binding of substrates in an inaccessible manner may cause the observed inhibitions. Overall, foam immobilization stabilizes enzymic activity and is a facile laboratory process with scaleup potential. PMID- 16347597 TI - Molecular Cloning and Expression of a Xylanase Gene from Bacillus polymyxa in Escherichia coli. AB - Genomic fragments of Bacillus polymyxa derived from separate and complete digestion by EcoRI, HindIII, and BamHI were ligated into the corresponding sites of pBR322, and the resulting chimeric plasmids were transformed into Escherichia coli. Of 6,000 transformants screened, 1 (pBPX-277) produced a clear halo on Remazol brilliant blue xylan plates. The insert in the pBPX-277 recombinant, identified as an 8.0-kilobase BamHI fragment of B. polymyxa, was subsequently subjected to extensive mapping and a series of subclonings into pUC19. A 2.9 kilobase BamHI-EcoRI subfragment was found to code for xylanase activity. Xylanase activity expressed by E. coli harboring the cloned gene was located primarily in the periplasm and corresponded to one of two distinct xylanases produced by B. polymyxa. Xylanase expression by the cloned gene occurred in the absence of xylan and was reduced by glucose and xylose. Southern blot hybridization with the cloned fragment as a probe against complete genomic digests of the bacilli B. polymyxa, B. circulans, and B. subtilis revealed that the cloned xylanase gene was unique to B. polymyxa. The xylanase expressed by the cloned gene had a molecular weight of approximately 48,000 and an isoelectric point of 4.9. PMID- 16347598 TI - Nitrilase-Catalyzed Production of Nicotinic Acid from 3-Cyanopyridine in Rhodococcus rhodochrous J1. AB - The nitrilase which occurs abundantly in cells of Rhodococcus rhodochrous J1 catalyzes the direct hydrolysis of 3-cyanopyridine to nicotinic acid without forming nicotinamide. By using resting cells, the reaction conditions for nicotinic acid production were optimized. Under the optimum conditions, 100% of the added 3-cyanopyridine could be converted to nicotinic acid, the highest yield achieved being 172 mg of nicotinic acid per 1.0 ml of reaction mixture containing 2.89 mg (dry weight) of cells in 26 h. PMID- 16347599 TI - Colony dimorphism in bradyrhizobium strains. AB - Ten isolates of Bradyrhizobium spp. which form two colony types were studied; the isolates originated from a range of legume species. The two colony types differed in the amount of gum formed or size or both, depending on the strain. Whole 7-day old colonies of each type were subcultured to determine the proportion of cells which had changed to the other type. An iterative computerized procedure was used to determine the rate of switching per generation between the two types and to predict proportions reached at equilibrium for each strain. The predicted proportions of the wetter (more gummy) or larger colony type at equilibrium differed significantly between strains, ranging from 0.9999 (strain CIAT 2383) to 0.0216 (strain CIAT 2469), because some strains switched faster from dry to wet (or small to large) and others switched faster from wet to dry (or large to small). Predicted equilibrium was reached after about 140 generations in strain USDA 76. In all but one strain (CIAT 3030) the growth rate of the wetter colony type was greater than or similar to that of the drier type. The mean difference in generation time between the two colony types was 0.37 h. Doubling times calculated for either colony type after 7 days of growth on the agar surface ranged from 6.0 to 7.3 h. The formation of two persistent colony types by one strain (clonal or colony dimorphism) may be a common phenomenon among Bradyrhizobium strains. PMID- 16347600 TI - Circular Permutation of the Genome of a Temperate Bacteriophage from Streptococcus cremoris BK5. AB - The temperate bacteriophage BK5-T was isolated from Streptococcus cremoris BK5 by induction with mitomycin C. Electron microscopy revealed that BK5-T DNA consists of linear molecules, ranging in size from 39.7 to 46 kilobase pairs. Restriction analysis of self-ligated BK5-T DNA showed that the ends of the DNA were not cohesive. The EcoRI restriction fragments of the phage genome were cloned into pACYC184. Restriction enzyme analysis of both the phage DNA and the cloned EcoRI fragments with EcoRI, BstEII, PstI, ClaI, and XbaI yielded a 37.6-kilobase-pair long circular restriction map for the phage genome. It was concluded that the BK5 T DNA molecules in the population differ in their sequence by a circular permutation and that individual DNA molecules are terminally redundant. The map location of the sites at which packaging of BK5-T DNA into phage heads is initiated (pac) and at which the phage integrates into the bacterial chromosome (att) were established. PMID- 16347601 TI - Comparative analysis of the chemical composition of mixed and pure cultures of green algae and their decomposed residues by C nuclear magnetic resonance spectroscopy. AB - It is known that macromolecular organic matter in aquatic environments, i.e., humic substances, is highly aliphatic. These aliphatic macromolecules, predominantly paraffinic in structure, are prevalent in marine and lacustrine sediments and are believed to originate from algae or bacteria. A comparative study of mixed and pure cultures of green algae and their decomposed residues was performed by using solid-state C nuclear magnetic resonance spectroscopy as the primary analytical method. Results obtained in this study confirm the presence of components that are chemically refractory and that are defined as alghumin and hydrolyzed alghumin. These were detected in heterogeneous, homogeneous, and axenic biomasses composed of several genera of Chlorophyta. Although the chemical composition of algal biomass varied with culture conditions, the chemical structure of the alghumin and hydrolyzed alghumin, demonstrated by C nuclear magnetic resonance spectroscopy appeared to be constant for members of the Chlorophyta examined in this study. The alghumin was dominated by carbohydrate carbon, with minor amounts of amide or carboxyl carbon and paraffinic carbon, the latter surviving strong hydrolysis by 6 N HCI (hydrolyzed alghumin). Bacterial decomposition of heterogeneous algal biomass labeled with C was conducted under both aerobic and anaerobic conditions to determine chemical structure and stability of the refractory material. The refractory fraction ranged from 33% in aerobic to 44% in anaerobic cultures. The refractory fraction recovered from either aerobic or anaerobic degradation comprised 40% alghumin, which represented an enrichment by 10% relative to the proportion of alghumin derived from whole cells of algae. The paraffinic component in the hydrolyzed alghumin of whole algal cells was found to be 1.8% and increased to 5.1 and 6.9% after aerobic and anaerobic bacterial degradation, respectively. It is concluded that members of the Chlorophyta contain a common insoluble structure composed of paraffinic carbon that is resistant to chemical and bacterial degradation under conditions used in this study. The paraffinic structure is identical to those constituting humin of aquatic origin. Thus, alga-derived macromolecular compounds deposited in aquatic environments (alghumin) probably contribute to sedimentary humic substances. PMID- 16347602 TI - Isolation of a new polysaccharide-digesting bacterium from a salt marsh. AB - A new marine bacterium that digested a variety of storage and structural polysaccharides, including agar, was isolated. Strain 2-40 is a nonfermentative gram-negative, polarly flagellated rod that sometimes grew as a filamentous helix and secreted a melaninlike pigment. Its characteristics conform to those of no previously described species. PMID- 16347603 TI - Effects of Metals on 3-Acetyldeoxynivalenol Production by Fusarium graminearum R2118 in Submerged Cultures. AB - The effects of selected metals (Mg, Mn, Zn, and Fe) on 3-acetyldeoxynivalenol (3 ADN) production by Fusarium graminearum R2118 and on its mycelial growth were investigated by using a two-stage submerged-culture technique. In certain concentrations ranges, Mg and Fe stimulated growth but suppressed 3-ADN production; at other concentrations, Mg, Fe, and Zn suppressed growth but stimulated 3-ADN production. In contrast, Mn stimulated growth but totally inhibited 3-ADN production at all concentrations tested. In general, the production of 3-ADN was inversely related to the growth rate of the fungus with these metals. Mn appears to be a crucial factor regulating the onset of 3-ADN biosynthesis. PMID- 16347604 TI - Isolation of Bacteria Which Can Grow at Both High pH and Low Temperature. AB - Fifty bacterial strains able to grow at pH 10 and 0 degrees C were isolated from soils, and growth characteristics of three selected strains were investigated. Strain 207, which showed the best growth rate of all the isolates at the conditions described above, could grow at a temperature of -5 to 39 degrees C at pH 8.5. The optimum pH for this strain changed from 9.5 at 10 degrees C to 9.0 at 20 degrees C. PMID- 16347606 TI - Poliovirus retention in soil columns after application of chemical- and polyelectrolyte-conditioned dewatered sludges. AB - [This corrects the article on p. 119 in vol. 54.]. PMID- 16347605 TI - Effect of a killer toxin of yeast on eucaryotic systems. AB - The Saccharomyces cerevisiae killer toxin KT 28, which inhibits sensitive yeasts, was shown to have no effect on several pathogenic fungi or on the protozoan Trichomonas vaginalis. At concentrations of about 0.1 mg/ml, a partial inhibition of the skin pathogenic fungi Trichophyton rubrum and Microsporum canis was observed at pH 6.5. No pharmacological activity was detected in various tests with several animal organs. PMID- 16347607 TI - Interference Competition among Coprophilous Fungi: Production of (+)-Isoepoxydon by Poronia punctata. AB - (+)-Isoepoxydon has been established as the major causative agent of interference competition between Poronia punctata (NRRL 6457), a late fungal colonist of cattle dung, and two early-occurring dung colonists, Ascobolus furfuraceus (NRRL 6460) and Sordaria fimicola (NRRL 6459). This compound was isolated from ethyl acetate extracts of liquid cultures of P. punctata by silica gel chromatography and identified by mass spectrometry and proton and carbon nuclear magnetic resonance spectroscopy. The isolation process was guided by in vitro bioassays for antifungal activity against A. furfuraceus and S. fimicola. (+)-Isoepoxydon has been implicated as an intermediate in the biosynthesis of patulin, a mycotoxin produced by Penicillium spp., but no patulin could be detected in cultures of P. punctata. PMID- 16347608 TI - Effect of Plasmid Incompatibility on DNA Transfer to Streptococcus cremoris. AB - Several Streptococcus cremoris strains were used in protoplast transformation and interspecific protoplast fusion experiments with Streptococcus lactis and Bacillus subtilis, with pGKV110, pGKV21, and DeltapAMbeta1 as the marker plasmids. DeltapAMbeta1 is a 15.9-kilobase nonconjugative, deletion derivative of pAMbeta1, which is considerably larger than the pGKV plasmids (approximately 4.5 kilobases). In general, DeltapAMbeta1 was transferred more efficiently than the pGKV plasmids. Using electroporation, we were able to demonstrate that failure of efficient transfer for the pGKV plasmids was, except for one case, caused by incompatibility of these plasmids with resident plasmids of the recipient strain. PMID- 16347609 TI - Development of Host Range Mutants of Xanthomonas campestris pv. translucens. AB - Xanthomonas campestris pv. translucens is the causal agent of bacterial leaf streak of cereal grains and grasses, and individual strains within the pathovar differ in their host range among the cereals. Coinoculation of a wide-host-range and a narrow-host-range strain resulted in the wide-host-range reaction. Transposon and chemical mutagenesis of the wide-host-range strain Xct4, pathogenic on barley, wheat, rye, and triticale, resulted in variants with reduced host range. When pathogenicity was inactivated independently for barley, wheat, triticale, and rye, wild-type symptoms were retained on the other members in the host range. Testing of some host range mutants on additional varieties of the cereals indicated some cultivar specificity. In addition, mutants nonpathogenic on combinations of the hosts or on all hosts were isolated. This suggests that there are independent positive factors determining host range in this species, rather than an avirulence gene system such as those determining race specificity in other plant pathogens. PMID- 16347610 TI - Inhibitory Effects of Methylcellulose on Cellulose Degradation by Ruminococcus flavefaciens. AB - Highly methylated, long-chain celluloses strongly inhibited cellulose degradation by several species of cellulolytic bacteria of ruminal origin. Specifically, the inhibitory effects of methylcellulose on the growth of Ruminococcus flavefaciens FD1 were concentration dependent, with complete inhibition at 0.1% (wt/vol). However, methylcellulose did not inhibit growth on cellobiose or cellulooligosaccharides. Mixtures of methylated cellulooligosaccharides having an average degree of polymerization of 6.7 to 9.5 inhibited cellulose degradation, but those with an average degree of polymerization of 1.0 to 4.5 did not. Similar inhibitory effects by methylcellulose and, to a lesser extent, by methyl cellulooligosaccharides were observed on cellulase activity, as measured by hydrolysis of p-nitrophenyl-beta-d-cellobioside. R. flavefaciens cultures hydrolyzed cellulooligosaccharides to cellobiose and cellotriose as final end products. Cellopentaose and cellohexaose were cleaved to these end products, but cellotetraose was also formed from cellohexaose. Methylcellulose did not inhibit hydrolysis of cellulooligosaccharides. These data are consistent with the presence of separate cellulase (beta-1,4-glucanase) and cellulodextrinase activities in R. flavefaciens. PMID- 16347611 TI - Influence of Storage at Freezing and Subsequent Refrigeration Temperatures on beta-Galactosidase Activity of Lactobacillus acidophilus. AB - The ability of three strains of Lactobacillus acidophilus to survive and retain beta-galactosidase activity during storage in liquid nitrogen at -196 degrees C and during subsequent storage in milk at 5 degrees C was tested. The level of beta-galactosidase activity varied among the three strains (0.048 to 0.177 U/10 organisms). Freezing and storage at -196 degrees C had much less adverse influence on viability and activity of the enzyme than did storage in milk at 5 degrees C. The strains varied in the extent of the losses of viability and beta galactosidase activity during both types of storage. There was not a significant interaction between storage at -196 degrees C and subsequent storage at 5 degrees C. The strains that exhibited the greatest losses of beta-galactosidase activity during storage in milk at 5 degrees C also exhibited the greatest losses in viability at 5 degrees C. However, the losses in viability were of much greater magnitude than were the losses of enzymatic activity. This indicates that some cells of L. acidophilus which failed to form colonies on the enumeration medium still possessed beta-galactosidase activity. Cultures of L. acidophilus to be used as dietary adjuncts to improve lactose utilization in humans should be carefully selected to ensure that adequate beta-galactosidase activity is provided. PMID- 16347612 TI - Ethanol-Induced Leakage in Saccharomyces cerevisiae: Kinetics and Relationship to Yeast Ethanol Tolerance and Alcohol Fermentation Productivity. AB - Ethanol stimulated the leakage of amino acids and 260-nm-light-absorbing compounds from cells of Saccharomyces cerevisiae. The efflux followed first-order kinetics over an initial period. In the presence of lethal concentrations of ethanol, the efflux rates at 30 and 36 degrees C were an exponential function of ethanol concentration: k(e) = k(e)e, where k(e) and k(e) are the efflux rate constants, respectively, in the presence of a concentration X of ethanol or the minimal concentration of ethanol, X(m), above which the equation was applicable, coincident with the minimal lethal concentration of ethanol. E is the enhancement constant. At 36 degrees C, as compared with the corresponding values at 30 degrees C, the efflux rates were higher and the minimal concentration of ethanol (X(m)) was lower. The exponential constants for the enhancement of the rate of leakage (E) had similar values at 30 or 36 degrees C and were of the same order of magnitude as the corresponding exponential constants for ethanol-induced death. Under isothermic conditions (30 degrees C) and up to 22% (vol/vol) ethanol, the resistance to ethanol-induced leakage of 260-nm-light-absorbing compounds was found to be closely related with the ethanol tolerance of three strains of yeasts, Kluyveromyces marxianus, Saccharomyces cerevisiae, and Saccharomyces bayanus. The resistance to ethanol-induced leakage indicates the possible adoption of the present method for the rapid screening of ethanol tolerant strains. The addition to a fermentation medium of the intracellular material obtained by ethanol permeabilization of yeast cells led to improvements in alcohol fermentation by S. cerevisiae and S. bayanus. The action of the intracellular material, by improving yeast ethanol tolerance, and the advantages of partially recycling the fermented medium after distillation were discussed. PMID- 16347613 TI - Selection of Ethanol-Tolerant Yeast Hybrids in pH-Regulated Continuous Culture. AB - Hybrids between naturally occurring wine yeast strains and laboratory strains were formed as a method of increasing genetic variability to improve the ethanol tolerance of yeast strains. The hybrids were subjected to competition experiments under continuous culture controlled by pH with increasing ethanol concentrations over a wide range to select the fastest-growing strain at any concentration of ethanol. The continuous culture system was obtained by controlling the dilution rate of a chemostat connected to a pH-meter. The nutrient pump of the chemostat was switched on and off in response to the pH of the culture, which was thereby kept near a critical value (pH(c)). Under these conditions, when the medium was supplemented with ethanol, the ethanol concentration of the culture increased with each pulse of dilution. A hybrid strain was selected by this procedure that was more tolerant than any of the highly ethanol-tolerant wine yeast strains at any concentration of ethanol and was able to grow at up to 16% (vol/vol) ethanol. This improvement in ethanol tolerance led to an increase in both the ethanol production rate and the total amount of ethanol produced. PMID- 16347614 TI - Two-stage mineralization of phenanthrene by estuarine enrichment cultures. AB - The polycyclic aromatic hydrocarbon phenanthrene was mineralized in two stages by soil, estuarine water, and sediment microbial populations. At high concentrations, phenanthrene was degraded, with the concomitant production of biomass and accumulation of Folin-Ciocalteau-reactive aromatic intermediates. Subsequent consumption of these intermediates resulted in a secondary increase in biomass. Analysis of intermediates by high-performance liquid chromatography, thin-layer chromatography, and UV absorption spectrometry showed 1-hydroxy-2 naphthoic acid (1H2NA) to be the predominant product. A less pronounced two-stage mineralization pattern was also observed by monitoring CO(2) production from low concentrations (0.5 mg liter) of radiolabeled phenanthrene. Here, mineralization of C-labeled 1H2NA could explain the incremental CO(2) produced during the later part of the incubations. Accumulation of 1H2NA by isolates obtained from enrichments was dependent on the initial phenanthrene concentration. The production of metabolites during polycyclic aromatic hydrocarbon biodegradation is discussed with regard to its possible adaptive significance and its methodological implications. PMID- 16347615 TI - Interaction of vesicular-arbuscular mycorrhizal fungi with erosion in an oxisol. AB - The development of vesicular-arbuscular mycorrhizal (VAM) symbiosis was monitored in Leucaena leucocephala grown in an Oxisol subjected to incremental simulated erosion. The density of VAM infective propagules in the soil diminished as the level of simulated erosion (removal of surface soil) was increased from 0 to 50 cm. The level of infection on L. leucocephala roots observed at harvest was not significantly influenced by simulated erosion unless removal of surface soil exceeded 25 cm. Inoculation of this soil and the uneroded soil with Glomus aggregatum enhanced the early onset of infection but did not significantly influence the level of infection observed at the time of harvest. Simulated erosion in excess of 7.5 cm of surface soil removal significantly delayed the development of VAM effectiveness monitored in terms of the P status of L. leucocephala subleaflets and also curtailed the level of maximum effectiveness observed. Decreases in VAM effectiveness were significantly correlated with decreases in soil chemical constituents. However, VAM effectiveness in a soil subjected to 30 cm of surface soil removal was not restored to a significant extent unless the soil was amended with P, even though other nutrients were restored to sufficiency levels. Our results demonstrate that the development of VAM effectiveness is the phase of the VAM symbiosis that is most adversely influenced by simulated erosion and that this effect appears to be caused primarily by insufficient P in the soil solution. PMID- 16347616 TI - Trichloroethylene biodegradation by a methane-oxidizing bacterium. AB - Trichloroethylene (TCE), a common groundwater contaminant, is a suspected carcinogen that is highly resistant to aerobic biodegradation. An aerobic, methane-oxidizing bacterium was isolated that degrades TCE in pure culture at concentrations commonly observed in contaminated groundwater. Strain 46-1, a type I methanotrophic bacterium, degraded TCE if grown on methane or methanol, producing CO(2) and water-soluble products. Gas chromatography and C radiotracer techniques were used to determine the rate, methane dependence, and mechanism of TCE biodegradation. TCE biodegradation by strain 46-1 appears to be a cometabolic process that occurs when the organism is actively metabolizing a suitable growth substrate such as methane or methanol. It is proposed that TCE biodegradation by methanotrophs occurs by formation of TCE epoxide, which breaks down spontaneously in water to form dichloroacetic and glyoxylic acids and one-carbon products. PMID- 16347617 TI - Transfer and occurrence of large mercury resistance plasmids in river epilithon. AB - In situ mating experiments were done in the River Taff, South Wales, United Kingdom, by using a natural mercury resistance plasmid (pQM1) isolated from a mixture of epilithic bacteria in vitro. The river temperature from March to November was found to influence transfer frequencies strongly (6.8 x 10 to 1.5 x 10 per recipient). A linear relationship existed between log(10) transfer frequency and river temperature (6 to 21 degrees C), a 2.6 degrees C change in temperature giving a 10-fold change in transfer frequency. In vitro experiments showed that pQM1 transferred most efficiently between fluorescent pseudomonads and that one epilithic isolate (Pseudomonas fluorescens) was an efficient donor in situ. Experiments with a P. putida recipient showed that intact epilithic bacterial communities could transfer mercury resistance plasmids in situ at frequencies of up to 3.75 x 10 per recipient. Nineteen of the large (>250 kilobase) plasmids isolated by transfer into P. putida were studied in detail and grouped into seven types by restriction digests. Mercury resistance and UV resistance were found to be common linked phenotypes in 19 of the 23 plasmids tested. PMID- 16347618 TI - Production and Purification of d-Aminoacylase from Alcaligenes denitrificans and Taxonomic Study of the Strain. AB - A d-aminoacylase-producing microorganism, strain DA181, isolated from soil was identified as Alcaligenes denitrificans subsp. denitrificans. This strain produced about 29,300 units (micromoles of product formed per hour) of d aminoacylase and 2,300 units of l-aminoacylase per gram of cells (wet weight) when cultivated in a medium containing 1% N-acetyl-dl-leucine as the carbon source. The d-aminoacylase was purified 345-fold. The specific activity of the purified enzyme was 108,600 units per mg of protein when N-acetyl-d-methionine was used as a substrate. The apparent molecular weight was 58,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. N-Acetyl-d methionine was the favored substrate, followed by N-acetyl-d-phenylalanine. This enzyme had a high stereospecificity, and its hydrolysis of N-acetyl-l-amino acids was almost negligible. PMID- 16347619 TI - Lysis of halobacteria in bacto-peptone by bile acids. AB - All tested strains of halophilic archaebacteria of the genera Halobacterium, Haloarcula, Haloferax, and Natronobacterium lysed in 1% Bacto-Peptone (Difco) containing 25% NaCl, whereas no lysis was observed with other strains belonging to archaebacteria of the genera Halococcus, Natronococcus, and Sulfolobus, methanogenic bacteria, and moderately halophilic eubacteria. Substances in Bacto Peptone which caused lysis of halobacteria were purified and identified as taurocholic acid and glycocholic acid. High-performance liquid chromatography analyses of peptones revealed that Bacto-Peptone contained nine different bile acids, with a total content of 9.53 mg/g, whereas much lower amounts were found in Peptone Bacteriological Technical (Difco) and Oxoid Peptone. Different kinds of peptones can be used to distinguish halophilic eubacteria and archaebacteria in mixed cultures from hypersaline environments. PMID- 16347620 TI - Strain Improvement of Rhodotorula graminis for Production of a Novel l Phenylalanine Ammonia-Lyase. AB - l-Phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) from Rhodotorula rubra has been used in the commercial manufacture of l-phenylalanine from trans-cinnamic acid. In this study, R. graminis PAL was investigated. Mutant strain GX6000 was isolated after ethyl methanesulfonate mutagenesis of wild-type R. graminis GX5007 by selecting for resistance to phenylpropiolic acid, an analog of trans-cinnamic acid. Mutant strain GX6000 produced inducible PAL at levels four- to fivefold higher than had wild-type R. graminis. Furthermore, this strain had several other physiological traits that make it more commercially useful than R. rubra. For example, during fermentation, the PAL half-life was three- to fivefold longer, PAL specific activity was six to seven times higher, and PAL synthesis was significantly less inhibited by temperatures above 30 degrees C. Induction of PAL in strain GX6000 appeared to be less tightly regulated; l-leucine acted synergistically with l-phenylalanine, the physiological inducer, to increase the PAL specific activity and titer to 165 U/g (dry weight) and 3,000 U/liter, respectively, a 40% increase over the effect of l-phenylalanine alone. Strain GX6000 PAL showed significantly greater stability in bioreactors for the synthesis of l-phenylalanine, a finding that is consistent with the stability properties observed during fermentation. PMID- 16347621 TI - Denitrification in a sand and gravel aquifer. AB - Denitrification was assayed by the acetylene blockage technique in slurried core material obtained from a freshwater sand and gravel aquifer. The aquifer, which has been contaminated with treated sewage for more than 50 years, had a contaminant plume greater than 3.5-km long. Near the contaminant source, groundwater nitrate concentrations were greater than 1 mM, whereas 0.25 km downgradient the central portion of the contaminant plume was anoxic and contained no detectable nitrate. Samples were obtained along the longitudinal axis of the plume (0 to 0.25 km) at several depths from four sites. Denitrification was evident at in situ nitrate concentrations at all sites tested; rates ranged from 2.3 to 260 pmol of N(2)O produced (g of wet sediment) h. Rates were highest nearest the contaminant source and decreased with increasing distance downgradient. Denitrification was the predominant nitrate reducing activity; no evidence was found for nitrate reduction to ammonium at any site. Denitrifying activity was carbon limited and not nitrate limited, except when the ambient nitrate level was less than the detection limit, in which case, even when amended with high concentrations of glucose and nitrate, the capacity to denitrify on a short-term basis was lacking. These results demonstrate that denitrification can occur in groundwater systems and, thereby, serve as a mechanism for nitrate removal from groundwater. PMID- 16347622 TI - Isolation and Characterization of Xylan-Degrading Strains of Butyrivibrio fibrisolvens from a Napier Grass-Fed Anaerobic Digester. AB - Six new xylanolytic bacterial strains have been isolated from a Napier grass-fed anaerobic digester. These strains were identified as Butyrivibrio fibrisolvens and were similar in many respects to ruminal isolates described previously. The new isolates exhibited a high degree of DNA homology with several ruminal strains of B. fibrisolvens. Xylan or xylose was required to induce the production of enzymes for xylan degradation, xylanase and xylosidase. Production of these enzymes was repressed in the presence of glucose. Xylanase activity was predominantly extracellular, while that of xylosidases was cell associated. The new isolates of B. fibrisolvens grew well in defined medium containing xylan as the sole carbon source and did not produce obvious slime or capsular layers. These strains may be useful for future genetic investigations. PMID- 16347623 TI - Rates of digestion of bacteria by marine phagotrophic protozoa: temperature dependence. AB - The effect of temperature on length of time for digestion of bacteria was evaluated, by using fluorescently labeled bacteria (FLB), for phagotrophic flagellates and ciliates isolated from coastal northwest Mediterranean waters. Accumulation of FLB in protozoan food vacuoles was followed until a plateau of FLB per cell occurred; then after a 1:10 dilution of FLB with unlabeled bacteria, disappearance of FLB in food vacuoles was monitored. For both 3- to 5-mum flagellates and 10- to 40-mum ciliates, the absolute linear slopes of FLB uptake and disappearance were nearly identical in individual experiments over a temperature range of 12 to 22 degrees C. We inferred from these results that the leveling off of the uptake curves resulted when equilibrium between ingestion and digestion of bacteria was attained. The time to leveling off then represented the average time needed for complete digestion of the bacteria ingested at the start of the experiment, and the inverse of this time represented a bacterial digestion rate. The digestion rate increased exponentially from 12 to 22 degrees C for both a mixed flagellate assemblage and the oligotrichous ciliate Strombidium sulcatum, with a Q(10) of 2.8 for the flagellates and 2.0 for the ciliate. Although bacterial ingestion rates varied greatly, depending on protozoan cell size, total bacterial abundance, and temperature, digestion times appeared to be significantly influenced only by protozoan cell size (or type of protozoan) and by temperature. PMID- 16347624 TI - Ecological indicators of native rhizobia in tropical soils. AB - The relationship between environment and abundance of rhizobia was described by determining the populations of root nodule bacteria at 14 diverse sites on the island of Maui. Mean annual rainfall at the sites ranged from 320 to 1,875 mm, elevation from 37 to 1,650 m, and soil pH from 4.6 to 7.9. Four different soil orders were represented in this study: inceptisols, mollisols, ultisols, and an oxisol. The rhizobial populations were determined by plant infection counts of five legumes (Trifolium repens, Medicago sativa, Vicia sativa, Leucaena leucocephala, and Macroptilium atropurpureum). Populations varied from 1.1 to 4.8 log(10) cells per g of soil. The most frequently occurring rhizobia were Bradyrhizobium spp., which were present at 13 of 14 sites with a maximum of 4.8 log(10) cells per g of soil. Rhizobium trifolii and R. leguminosarum occurred only at higher elevations. The presence of a particular Rhizobium or Bradyrhizobium sp. was correlated with the occurrence of its appropriate host legume. Total rhizobial populations were significantly correlated with mean annual rainfall, legume cover and shoot biomass, soil temperature, soil pH, and phosphorus retention. Regression models are presented which describe the relationship of legume hosts, soil climate, and soil fertility on native rhizobial populations. PMID- 16347625 TI - Relationship between Methanogenic Cofactor Content and Maximum Specific Methanogenic Activity of Anaerobic Granular Sludges. AB - In this study we investigated whether a relationship exists between the methanogenic activity and the content of specific methanogenic cofactors of granular sludges cultured on different combinations of volatile fatty acids in upflow anaerobic sludge blanket or fluidized-bed reactors. Significant correlations were measured in both cases between the contents of coenzyme F(420) 2 or methanopterin and the maximum specific methanogenic activities on propionate, butyrate, and hydrogen, but not acetate. For both sludges the content of sarcinapterin appeared to be correlated with methanogenic activities on propionate, butyrate, and acetate, but not hydrogen. Similar correlations were measured with regard to the total content of coenzyme F(420)-4 and F(420)-5 in sludges from fluidized-bed reactors. The results indicate that the contents of specific methanogenic cofactors measured in anaerobic granular sludges can be used to estimate the hydrogenotrophic or acetotrophic methanogenic potential of these sludges. PMID- 16347626 TI - Production, Distribution, and Kinetic Properties of Inulinase in Continuous Cultures of Kluyveromyces marxianus CBS 6556. AB - From a screening of several Kluyveromyces strains, the yeast Kluyveromyces marxianus CBS 6556 was selected for a study of the parameters relevant to the commercial production of inulinase (EC 3.2.1.7). This yeast exhibited superior properties with respect to growth at elevated temperatures (40 to 45 degrees C), substrate specificity, and inulinase production. In sucrose-limited chemostat cultures growing on mineral medium, the amount of enzyme decreased from 52 U mg of cell dry weight at D = 0.1 h to 2 U mg of cell dry weight at D = 0.8 h. Experiments with nitrogen-limited cultures further confirmed that synthesis of the enzyme is negatively controlled by the residual sugar concentration in the culture. High enzyme activities were observed during growth on nonsugar substrates, indicating that synthesis of the enzyme is a result of a derepression/repression mechanism. A substantial part of the inulinase produced by K. marxianus was associated with the cell wall. The enzyme could be released from the cell wall via a simple chemical treatment of cells. Results are presented on the effect of cultivation conditions on the distribution of the enzyme. Inulinase was active with sucrose, raffinose, stachyose, and inulin as substrates and exhibited an S/I ratio (relative activities with sucrose and inulin) of 15 under standard assay conditions. The enzyme activity decreased with increasing chain length of the substrate. PMID- 16347627 TI - Characterization of Lactobacillus bulgaricus Bacteriophage ch2. AB - Bacteriophage ch2, a virulent bacteriophage of Lactobacillus bulgaricus CH2, was characterized according to its morphology, genome size, structural proteins, and growth kinetics. Electron micrographs revealed that bacteriophage ch2 has an icosahedral head of 50-nm diameter and a long tail of 170 nm. Its genome is linear and 35 kilobases in length, and its structural proteins consist of two major and eight minor proteins. One-step growth kinetics of bacteriophage ch2 under optimal conditions (45 degrees C in MRS medium [Oxoid Ltd.]) showed that the latent time was 40 min, the rise period was 15 min, and the burst size was 130 bacteriophages per cell. To monitor the effects of bacteriophage infection on host growth and beta-galactosidase production, the absorbance of the culture and the beta-galactosidase activity were followed during the infection cycle. Before lysis the infected culture continued to grow and produce beta-galactosidase at the same rate as the uninfected culture. PMID- 16347628 TI - Populations of methanogenic bacteria in a georgia salt marsh. AB - Methanogens represented about 0.5% of the total bacteria in sediments from a Georgia salt marsh in which Spartina alterniflora is the predominant vegetation. The population of methanogens was composed of at least two groups of nearly equal size. One group was represented by cocci which were able to utilize trimethylamine and were unable to use H(2) or acetate. The second group was composed of two subgroups which were able to utilize H(2) but were unable to use trimethylamine or acetate. The more common subgroup included rod- or plate-shaped methanogens which could utilize isopropanol in addition to H(2) and formate. The second subgroup included Methanococcus maripaludis, which utilized only H(2) and formate. Other groups of methanogens were also present, including Methanosarcina sp. which utilized acetate, H(2), and methylamines. In addition to the overall variability in the types of methanogens, the numbers of methanogens in sediments also exhibited significant spatial variability both within and between tall- and short-Spartina zones. PMID- 16347629 TI - Ferulic Acid Esterase Activity from Schizophyllum commune. AB - Schizophyllum commune produced an esterase which released ferulic acid from starch-free wheat bran and from a soluble ferulic acid-sugar ester that was isolated from wheat bran. The preferred growth substrate for the production of ferulic acid esterase was cellulose. Growth on xylan-containing substrates (oat spelt xylan and starch-free wheat bran) resulted in activity levels that were significantly lower than those observed in cultures grown on cellulose. Similar observations were made for endoglucanase, p-nitrophenyllactopyranosidase, xylanase, and acetyl xylan esterase. Of the enzymes studied, only arabinofuranosidase was produced at maximum levels during growth on xylan containing materials. Ferulic acid esterase that had been partially purified by DEAE chromatography released significant amounts of ferulic acid from wheat bran only in the presence of a xylanase-rich fraction, indicating that the esterase may not be able to readily attack high-molecular-weight substrates. The esterase acted efficiently, without xylanase addition, on a soluble sugar-ferulic acid substrate. PMID- 16347630 TI - Microbial Ecology of the Gut in Laboratory Stocks of the Migratory Grasshopper, Melanoplus sanguinipes (Fab.) (Orthoptera: Acrididae). AB - Mean pH values in pooled samples of foregut, midgut, and hindgut from adult Melanoplus sanguinipes, which had been raised in the laboratory on barley shoots and wheat bran, were 5.15, 6.39, and 5.98, respectively. Homogenates of midgut/hindgut sections and frass (feces) yielded colony counts of bacteria by the spread plate method of 5.7 to 5.9 and 5.3 to 5.5 log(10) colonies per mg, respectively; there were no significant differences (P > 0.05) between counts obtained on several media or on media incubated aerobically or anaerobically. There was no evidence of significant populations of protozoa, fungi, or obligately anaerobic bacteria associated with the gut. A total of 168 pure strains of bacteria isolated from the gut sections were characterized and assigned to 11 taxonomic groups, including Enterococcus spp., Serratia liquefaciens, Pseudomonas spp., and Enterobacter spp. Numbers of Enterococcus spp. in the gut were 2 to 3 orders of magnitude higher than those of the other genera. Strains representing only four of the groups were recovered from bran fed to the grasshoppers; the barley shoots, which were raised in sterile soil, appeared virtually sterile. Examination of the gut wall by scanning electron microscopy revealed the presence of epimural bacteria in the foregut and hindgut but not in the midgut. The distribution of epimural cocci and bacilli differed with the gut section examined. Numerous spherical to ovoid structures up to 10 mum in diameter, which were not identified, were associated with the microvillous surface of the midgut epithelium. Acetate was present in gut, hemolymph, and frass, and it was shown that representative isolates of Enterococcus spp. and Enterobacter agglomerans produced acetate when incubated in an aqueous suspension of bran. The egestion time of solid digesta, as measured with methylene blue stained barley shoots, was 3.0 to 5.7 h. The results show that M. sanguinipes supported extensive indigenous populations of luminal and epimural bacteria in the gut which were composed predominantly of facultatively anaerobic species; the relatively short egestion time, indicating rapid passage of digesta through the gut, was consistent with the microscopic appearance of digesta residues in frass and could account, at least in part, for the absence of a significant population of obligately anaerobic bacteria from the gut. PMID- 16347631 TI - Extremely thermophilic fermentative archaebacteria of the genus desulfurococcus from deep-sea hydrothermal vents. AB - Two strains of extremely thermophilic, anaerobic bacteria are described that are representative of isolates obtained from a variety of oceanic hydrothermal vent sites at depths from 2,000 to 3,700 m. The isolates were similar in their requirements for complex organic media, elemental sulfur, and seawater-range salinities (optimum, 2.1 to 2.4%); their high tolerance for sulfide (100 mM) and oxic conditions below growth-range temperatures (50 to 95 degrees C); and their archaebacterial characteristics: absence of murein, presence of certain diand tetraethers, and response to specific antibiotics. The two strains (S and SY, respectively) differed slightly in their optimum growth temperatures (85 and 90 degrees C, optimum pHs for growth (7.5 and 7.0), and DNA base compositions (52.01 and 52.42 G+C mol%). At their in situ pressure of about 250 atm (25,313 kPa), growth rates at 80 and 90 degrees C were about 40% lower than those at 1 atm (101.29 kPa), and no growth occurred at 100 and 110 degrees C, respectively, at either pressure. In yeast extract medium, only 2% of the organic carbon was used and appeared to stem largely from the proteinaceous constituents. According to physiological criteria, the isolates belong to the genus Desulfurococcus. PMID- 16347632 TI - Effects of Stirring and Hydrogen on Fermentation Products of Clostridium thermocellum. AB - Clostridium thermocellum produces ethanol, acetate, H(2), and CO(2) as major fermentation products from cellulose and cellobiose. The performance of three strains of this microorganism was studied to assess the potential use in producing ethanol directly from cellulosic fiber. Depending on the bacterial strain, an ethanol/acetate product ratio from 1 to as high as 3 was observed in unstirred cultures. Vigorous stirring during growth resulted in a threefold decrease in the ethanol/acetate ratio. The H(2) content in the unstirred culture broth was three times greater than that in the stirred one. Addition of exogenous H(2) to the gas phase during growth increased the ethanol/acetate ratio much more in the stirred than in the unstirred fermentations. The addition of sufficient H(2) to the gas phase almost relieved the effect of stirring, and the ethanol/acetate ratio approached that in the unstirred condition. Addition of tritium to the gas phase of the culture resulted in the formation of tritiated water (H(2)O), which indicates that C. thermocellum possesses hydrogenase(s) that catalyzes the reverse reaction. The rate of H(2)O formation was about three times higher in the stirred culture than in the unstirred culture. These results demonstrate that the H(2) concentration in the broth plays an important role in the product formation. The H(2) supersaturation present in the unstirred cultures is responsible for the observed effect of stirring. A hydrogen feedback control mechanism regulating the relative concentrations of reduced and oxidized electron carriers is proposed to account for the effect of hydrogen on the metabolite distribution. PMID- 16347633 TI - Role of antibiosis in competition of erwinia strains in potato infection courts. AB - Erwinia carotovora subsp. betavasculorum strains produced a bactericidal antibiotic in vitro that inhibited a wide spectrum of gram-negative and gram positive bacteria. The optimum temperature for production was 24 degrees C, and the addition of glycerol to culture media enhanced antibiotic production. Antibiotic production by these strains in the infection court of potato was the principal determinant enabling it to gain ascendancy over competing antibiotic sensitive Erwinia carotovora subsp. carotovora strains. There was a complete correlation between antibiotic production by E. carotovora subsp. betavasculorum in vitro and inhibition of competing E. carotovora subsp. carotovora strains in planta. Inhibition of the latter by the former was apparent after 10 h of incubation in potato tuber wounds. Population densities of sensitive E. carotovora subsp. carotovora strains in mixed potato tuber infections with E. carotovora subsp. betavasculorum were approximately 10-fold lower after 48 h of incubation than in corresponding single sensitive strain infections. E. carotovora subsp. carotovora were not inhibited in tuber infections that were incubated anaerobically. This correlated with the absence of antibiotic production during anaerobic incubation in vitro. Antibiotic-resistant strains of E. carotovora subsp. carotovora were not inhibited in planta or in vitro by E. carotovora subsp. betavasculorum. Moreover, isogenic antibiotic-negative (Ant) mutant E. carotovora subsp. betavasculorum strains were not inhibitory to sensitive E. carotovora subsp. carotovora strains in tuber infections. PMID- 16347634 TI - Intergeneric Protoplast Fusion between Ruminococcus albus and an Anaerobic Recombinant, FE7. AB - Intergeneric protoplast fusion between Ruminococcus albus, a cellulolytic, gram positive, anaerobic bacterium (Pc Sm Km), and an anaerobic recombinant, FE7 (Pc Sm Km), having lignin-related compound-degrading activities, was performed under strictly anaerobic conditions to introduce cellulase genes into strain FE7. The fusion frequency varied with different selected markers from 3.0 x 10 to 3.3 x 10. Two fusants, obtained from a synthetic medium with selective pressures of penicillin and streptomycin and with cellooli-gomer as the sole carbon source, were gram-negative rods. One of them, named FE7R2, showed 45 to 47% of the beta glucosidase and cellobiosidase activities of its parent R. albus and still maintained a level of degradation activity against dehydrodivanillin, a lignin related compound, of up to 87% of that of the parent strain FE7. To verify that the cellulolytic activities expressed in the fusant FE7R2 originated from R. albus cellulase genes, the beta-glucosidase gene of R. albus was cloned into Escherichia coli HB101 with plasmid pBR322. Cells bearing a recombinant plasmid, pRAII, produced high enzyme activities against both p-nitrophenyl-beta-d glucoside and p-nitrophenyl-beta-d-cellobioside and could degrade cellobiose to glucose. Southern blot results showed that the cloned DNA fragment could hybridize with chromosomal DNAs of both R. albus and FE7R2, but did not with the chromosomal DNA of FE7, indicating that the beta-glucosidase gene fragment was introduced into the chromosome of FE7R2 from R. albus via the protoplast fusion. The fusant FE7R2 could utilize simultaneously both cellobiose and dehydrodivanillin. These results gave evidence that the fusion product FE7R2 is a recombinant strain between its parents R. albus and FE7. This recombinant has stably kept the above properties for about 2 years. PMID- 16347635 TI - High Pressure Enhances the Growth Rate of the Thermophilic Archaebacterium Methanococcus thermolithotrophicus without Extending Its Temperature Range. AB - Temperature and hydrostatic pressure are essential in determining the assemblage of species in their specific biotopes. To evaluate the effect of high pressure on the range of viability of thermophiles, the pressure and temperature dependence of the growth of the methanogenic archaebacterium Methanococcus thermolithotrophicus was investigated. High pressure up to 50 MPa enhanced the growth rate without extending the temperature range of viability. The optimum temperature remained unaltered (65 degrees C). Beyond 50 MPa, cell lysis predominated over cell proliferation. Destabilization was also observed at temperatures below and above the optimum growth temperature (<60 degrees C, >/=70 degrees C) and at low substrate concentrations. PMID- 16347636 TI - Diversity of Plasmid Profiles and Conservation of Symbiotic Nitrogen Fixation Genes in Newly Isolated Rhizobium Strains Nodulating Sulla (Hedysarum coronarium L.). AB - Forty-five Rhizobium strains nodulating sulla (Hedysarum coronarium L.), isolated from plants grown in different sites in Menorca Island and southern Spain, were examined for plasmid content and the location and organization of nif (nitrogen fixation) and nod (nodulation) sequences. A great diversity in both number and size of the plasmids was observed in this native population of strains, which could be distributed among 19 different groups according to their plasmid profiles. No correlation was found between plasmid profile and geographical origin of the strains. In each strain a single plasmid ranging from 187 to 349 megadaltons hybridized to Rhizobium meliloti nifHD and nodD DNA, and in three strains the spontaneous loss of this plasmid resulted in the loss of the nodulation capacity. In addition to the symbiotic plasmid, 18 different cryptic plasmids were identified. A characteristic cryptic plasmid of >1,000 megadaltons was present in all strains. Total DNA hybridization experiments, with nifHD and portions of nodC and nodD genes (coding for common nodulation functions) from R. meliloti as probes, demonstrated that both the sequence and organization of nif and common nod genes were highly conserved within rhizobia nodulating sulla. Evidence for reiteration of nodD sequences and for linkage of nodC to at least one copy of nodD was obtained for all the strains examined. From these results we conclude that Rhizobium strains nodulating sulla are a homogeneous group of symbiotic bacteria that are closely related to the classical fast-growing group of rhizobia. PMID- 16347637 TI - Narrow- and Broad-Host-Range Symbiotic Plasmids of Rhizobium spp. Strains That Nodulate Phaseolus vulgaris. AB - Agrobacterium transconjugants containing symbiotic plasmids from different Rhizobium spp. strains that nodulate Phaseolus vulgaris were obtained. All transconjugants conserved the parental nodulation host range. Symbiotic (Sym) plasmids of Rhizobium strains isolated originally from P. vulgaris nodules, which had a broad nodulation host range, and single-copy nitrogenase genes conferred a Fix phenotype to the Agrobacterium transconjugants. A Fix phenotype was obtained with Sym plasmids of strains isolated from P. vulgaris nodules that had a narrow host range and reiterated nif genes, as well as with Sym plasmids of strains isolated from other legumes that presented single nif genes and a broad nodulation host range. This indicates that different types of Sym plasmids can confer the ability to establish an effective symbiosis with P. vulgaris. PMID- 16347638 TI - Frequency and characteristics of plasmids in bacteria isolated from deep-sea amphipods. AB - Bacterial strains isolated from deep-sea amphipods were identified, classified, and screened for plasmid content. Plasmids were common, with 11 of 16 isolates carrying one or more plasmids; these ranged in size from 2.9 to 63 megadaltons. Several of the strains demonstrated distinctly different phenotypic traits yet contained plasmids of the same molecular weight. Results of agarose gel electrophoresis, DNA hybridization, and restriction analysis indicate that the plasmids detected in these deep-sea isolates are identical, suggesting that transmission may occur in the deep-sea environment and that plasmids are common in some deep-sea habitats. PMID- 16347639 TI - Degradation of the Phosphonate Herbicide Glyphosate by Arthrobacter atrocyaneus ATCC 13752. AB - Of nine authentic Arthrobacter strains tested, only A. atrocyaneus ATCC 13752 was capable of using the herbicide glyphosate [N-(phosphonomethyl)glycine] as its sole source of phosphorus. Contrary to the previously isolated Arthrobacter sp. strain GLP-1, which degrades glyphosate via sarcosine, A. atrocyaneus metabolized glyphosate to aminomethylphosphonic acid. The carbon of aminomethylphosphonic acid was entirely converted to CO(2). This is the first report on glyphosate degradation by a bacterial strain without previous selection for glyphosate utilization as a source of phosphorus. PMID- 16347640 TI - A proposed pathway for catabolism of propionate in methanogenic cocultures. AB - A metabolic pathway for the catabolism of propionate is proposed. This pathway incorporates a transcarboxylation reaction involving propionyl coenzyme A and oxaloacetate and a carboxylation of pyruvate to regenerate oxaloacetate. Data indicated that the proposed pathway is reversible. The proposed pathway and its apparent reversibility provide a reasonable explanation of observations obtained from metabolism of labeled substrate. PMID- 16347641 TI - One-Step Purification of a Lipolytic Enzyme from Bacillus pumilus by Mild Hydrophobic Interaction Chromatography on Polyoxyalkylene Glycol-Bound Sepharose 6B. AB - An esterase from Bacillus pumilus was obtained in a highly purified form, starting from a crude extract, by a new chromatographic technique requiring no detergent during the elution step. The stationary phase involved was mildly hydrophobic and was prepared by covalent immobilization of a polyoxyalkylene glycol onto Sepharose 6B. PMID- 16347642 TI - Transfer of Plasmids between Bacillus subtilis and Streptococcus lactis. AB - The shuttle plasmid pGK12, as well as several Staphylococcus aureus plasmids, was introduced into Streptococcus lactis by intergeneric protoplast fusion with Bacillus subtilis. The S. aureus plasmids were stably inherited in S. lactis, and so they may possibly be used as cloning vectors in lactic streptococci. PMID- 16347643 TI - Enumeration of free-living aerobic n(2)-fixing h(2)-oxidizing bacteria by using a heterotrophic semisolid medium and most-probable-number technique. AB - A heterotrophic semisolid medium was used with two sensitive assay methods, C(2)H(2) reduction and O(2)-dependent tritium uptake, to determine nitrogenase and hydrogenase activities, respectively. Organisms known to be positive for both activities showed hydrogenase activity in both the presence and absence of 1% C(2)H(2), and thus, it was possible to test a single culture for both activities. Hydrogen uptake activity was detected for the first time in N(2)-fixing strains of Pseudomonas stutzeri. The method was then applied to the most-probable-number method of counting N(2)-fixing and H(2)-oxidizing bacteria in some natural systems. The numbers of H(2)-oxidizing diazotrophs were considerably higher in soil surrounding nodules of white beans than they were in the other systems tested. This observation is consistent with reports that the rhizosphere may be an important ecological niche for H(2) transformation. PMID- 16347644 TI - Evaluation of the Role of Syringomycin in Plant Pathogenesis by Using Tn5 Mutants of Pseudomonas syringae pv. syringae Defective in Syringomycin Production. AB - Syringomycin is a necrosis-inducing phytotoxin produced by Pseudomonas syringae pv. syringae. To determine whether syringomycin production is a determinant in virulence or pathogenicity, we isolated nontoxigenic (Tox) Tn5-containing mutants and then quantitatively evaluated them for the ability to multiply and cause disease in immature sweet-cherry fruits. Transposon Tn5 was delivered to Tox strain B301D-R by using the suicide vector, pGS9, and the resultant kanamycin resistant (Km) colonies were screened for changes in syringomycin production by testing for antibiosis against Geotrichum candidum. Southern blot analysis of KpnI-and EcoRI-digested DNA showed that 15 (0.3%) Tox mutants were isolated which had Tn5 inserted into 1 of 14 distinct loci. Phenotypic characterization of the Tox mutants identified three major groups, which were differentiated by pathogenicity and ability to cause a tobacco hypersensitive reaction (HR). The eight strains in group A were pathogenic (Path) in cherry fruit assays, but the disease index was 17 to 66% lower (significant at P = 0.01) than for the parental Tox strain, B301D-R. The population dynamics of group A strains W4S770 and W4S116 in cherry fruits were, however, indistinguishable from that of strain B301D-R. The remaining seven Tox strains were nonpathogenic; group B strain W4S2545 (Path HR) and group C strain W4S468 (Path HR) developed significantly lower populations (10 to 10 CFU per cherry fruit) 3 days after inoculation than strain B301D-R did (nearly 10 CFU per fruit). The data indicate that syringomycin is not essential for pathogenicity, but contributes significantly to virulence. PMID- 16347645 TI - Bioenergetic conditions of butyrate metabolism by a syntrophic, anaerobic bacterium in coculture with hydrogen-oxidizing methanogenic and sulfidogenic bacteria. AB - The butyrate-oxidizing, proton-reducing, obligately anaerobic bacterium NSF-2 was grown in batch cocultures with either the hydrogen-oxidizing bacterium Methanospirillum hungatei PM-1 or Desulfovibrio sp. strain PS-1. Metabolism of butyrate occurred in two phases. The first phase exhibited exponential growth kinetics (phase a) and had a doubling time of 10 h. This value was independent of whether NSF-2 was cultured with a methanogen or a sulfate reducer and likely represents the maximum specific growth rate of NSF-2. This exponential growth phase was followed by a second phase with a nearly constant rate of degradation (phase b) which dominated the time course of butyrate degradation. The specific activity of H(2) uptake by the hydrogen-oxidizing bacterium controlled the bioenergetic conditions of metabolism in phase b. During this phase both the Gibbs free energy (DeltaG') and the butyrate degradation rate (v) were greater for NSF-2-Desulfovibrio sp. strain PS-1 (DeltaG' = -17.0 kJ/mol; v = 0.20 mM/h) than for NSF-2-M. hungatei PM-1 (DeltaG' = -3.8 kJ/mol, v = 0.12 mM/h). The DeltaG' value remained stable and characteristic of the two hydrogen oxidizers during phase b. The stable DeltaG' resulted from the close coupling of the rates of butyrate and H(2) oxidation. The addition of 2-bromoethanesulfonate to a NSF-2 methanogen coculture resulted in the total inhibition of butyrate degradation; the inhibition was relieved when Desulfovibrio sp. strain PS-1 was added as a new H(2) sink. When the specific activity of H(2) consumption was increased by adding higher densities of the Desulfovibrio sp. to 2-bromoethanesulfonate-inhibited NSF 2-methanogen cocultures, lower H(2) pool sizes and higher rates of butyrate degradation resulted. Thus, it is the kinetic parameters of H(2) consumption, not the type of H(2) consumer per se, that establishes the thermodynamic conditions which in turn control the rate of fatty acid degradation. The bioenergetic homeostasis we observed in phase b was a result of the kinetics of the coculture members and the feedback inhibition by hydrogen which prevents butyrate degradation rates from reaching their theoretical V(max). PMID- 16347646 TI - Populations of Anaerobic Phototrophic Bacteria in a Spartina alterniflora Salt Marsh. AB - Habitat-simulating media were used with the Hungate anaerobic roll tube technique to enumerate culturable anaerobic photosynthetic bacteria in sediment, tidal waters, and Spartina alterniflora plant samples collected from the salt marsh at Sapelo Island, Ga. No phototrophs were detected in samples of creekside (low marsh) sediment or in tidal waters in creekside regions. In the high marsh region, 90% of anaerobic phototrophic bacteria occurred in the top 5 mm of sediment and none were detected below 6 mm. There was a seasonal variation, with maximal populations occurring in summer and fall (mean, 4.4 x 10 phototrophs g of dry sediment) and minimal numbers occurring in winter (mean, 3.9 x 10 phototrophs g of dry sediment). During winter and late spring, phototrophs had a patchy distribution over the high marsh sediment surface. In contrast, during late summer they had a random uniform distribution. Tidal water collected over high marsh sediment contained an average of 8.7 x 10 phototrophs ml, with no significant seasonal variation. Anaerobic phototrophic bacteria were also cultured from the lower stem tissue of S. alterniflora growing in both the high (4.3 x 10 phototrophs g of dry tissue) and creekside (4.9 x 10 phototrophs g of dry tissue) marsh regions. Chromatium buderi, Chromatium vinosum, Thiospirillum sanguineum, Rhodospirillum molischianum, and Chlorobium phaeobacteroides were the predominant anaerobic phototrophic species cultured from high marsh sediment. The two Chromatium species were dominant. PMID- 16347647 TI - Plugging of a model rock system by using starved bacteria. AB - The effects of starvation on bacterial penetration through artificial rock cores were examined. Klebsiella pneumoniae was starved in a simple salts solution for a duration of up to 4 weeks. These cell suspensions were injected into sintered glass bead cores, and the resulting reductions in core permeabilities were recorded. Vegetative cell cultures of K. pneumoniae grown in a sodium citrate medium were injected into other, similar cores, and the reductions in core permeabilities were recorded. The starved cell suspensions did not completely block the core pores, whereas the vegetative cultures reduced core permeability to less than 1%. Scanning electron microscopy of core sections infiltrated with either vegetative or starved cells showed that the former produced shallow "skin" plugs and copious amounts of glycocalyx at the inlet face, whereas the latter produced very little glycocalyx and the cells were distributed evenly throughout the length of the core. The use of a DNA assay to produce a cell distribution profile showed that, compared with the vegetative cells, starved bacteria were able to penetrate deeper into the cores. This was due to the smaller size of the cells and the reduction in biofilm production. This ability of starved bacteria to penetrate further into cores than the normal-size vegetative cells can be usefully applied to selective plugging for enhanced oil recovery. To further test the suitability of starved cells for use in selective plugging, the activities of starved cells present within cores were monitored before and after nutrient stimulation. Our data indicate that with nutrient stimulation, the starved cells lose their metabolic dormancy and produce reductions in core permeability due to cell growth and polymer production. PMID- 16347648 TI - Nutrient resuscitation and growth of starved cells in sandstone cores: a novel approach to enhanced oil recovery. AB - Klebsiella pneumoniae, which was reduced in size (0.25 by 0.5 mum) by carbon deprivation, was injected into a series of sandstone cores and subjected to separate treatments. Scanning electron microscopy of 400-mD cores showed these small starved cells in nearly every core section. The cells were a mixture of small rods and cocci with little or no biofilm production. Continuous or dose stimulation with sodium citrate allowed the cells to grow throughout the sandstone and completely plug the length of the core. The resuscitated cells were larger than the starved cells (up to 1.7 mum) and were encased in glycocalyx. Scanning electron microscopic results of resuscitation in situ with half-strength brain heart infusion broth showed that a shallow "skin" plug of cells formed at the core inlet and that fewer cells were located in the lower sections. Starved cells also penetrated 200-mD cores and were successfully resuscitated in situ with sodium citrate, so that the entire core was plugged. Nutrient resuscitation of injected starved cells to produce full-size cells which grow and block the rock pores may be successfully applied to selective plugging and may effectively increase oil recovery. PMID- 16347649 TI - Microbial biomass, activity, and community structure of water and particulates retrieved by backflow from a waterflood injection well. AB - Oil field injection water was allowed to back flow from two wells at the Packard drill site in Los Angeles, Calif., and was sampled at various times to obtain information about the biomass, potential activity, and community structure of the microbiota in the reservoir formation and in the injection water. Biomass was greatest in water samples that came from the zone near the injection site and dropped off sharply in subsequent samples, which were assumed to come from zones farther away from the well. Samples obtained from near the well also had visible exopolysaccharide blankets, as seen in scanning electron microscopic preparations. In one of the wells that was sampled, rates of glucose or acetate incorporation into microbial lipids correlated with biomass; but in the other well, activities correlated with the sampling time (volume of water that back flowed). Transmission electron micrographs showed a diverse, gram-negative bacterial population in a variety of physiological states. The analysis of the phospholipid ester-linked fatty acid profiles of the samples revealed consistently large proportions of 18:1omega7c fatty acids, indicating the presence of many anaerobes, facultative organisms, or both. Proportions of cyclopropyl fatty acids and ratios of trans/cis monoenoic compounds increased with the volume of water that back flowed (analogous with the distance into the formation), while the ratio of unsaturated/saturated compounds decreased, possibly indicating higher levels of stress or starvation in the microbial communities farthest from the injection well. Greater than 90% of the total biomass was trapped on glass fiber filters, indicating that the microbiota were largely attached to particles or were clumped. These sampling techniques and analytical methods may prove useful in monitoring for problems with microbes (e.g., plugging) in waterflood operations and in the preparation of water injection wells for enhanced oil recovery by the use of microbes. PMID- 16347650 TI - Steady-State Effects of 2,5,2',5'-Tetrachlorobiphenyl on Growth, Photosynthesis, and P Uptake in Selenastrum capricornutum. AB - The steady-state effect of 2,5,2',5'-tetrachlorobiphenyl (TCBP) on the green alga Selenastrum capricornutum was investigated in a P-limited two-stage chemostat system. The partition coefficient of this polychlorinated biphenyl congener was 5.9 x 10 in steady-state cultures. At a cellular TCBP concentration of 12.2 x 10 ng . cell, growth rate was not affected. However, photosynthetic capacity (P(max)) was significantly enhanced by TCBP (56 x 10 mumol of C . cell . h versus 34 x 10 mumol of C . cell . h in the control). Photosynthetic efficiency, or the slope of the photosynthesis-irradiance curve, was also significantly higher. There was little difference in the cell chlorophyll a content, and therefore the difference in these photosynthetic characteristics was the same even when they were expressed on a per-chlorophyll a basis. Cell C content was higher in TCBP containing cells than in TCBP-free cells, but approximately 36% of the C fixed by cells with TCBP was not incorporated as cell C. The maximum P uptake rate was also enhanced by TCBP, but the half-saturation concentration appeared to be unaffected. PMID- 16347651 TI - Sub-Parts-Per-Billion Nitrate Method: Use of an N(2)O-Producing Denitrifier to Convert NO(3) or NO(3) to N(2)O. AB - A more sensitive analytical method for NO(3) was developed based on the conversion of NO(3) to N(2)O by a denitrifier that could not reduce N(2)O further. The improved detectability resulted from the high sensitivity of the Ni electron capture gas chromatographic detector for N(2)O and the purification of the nitrogen afforded by the transformation of the N to a gaseous product with a low atmospheric background. The selected denitrifier quantitatively converted NO(3) to N(2)O within 10 min. The optimum measurement range was from 0.5 to 50 ppb (50 mug/liter) of NO(3) N, and the detection limit was 0.2 ppb of N. The values measured by the denitrifier method compared well with those measured by the high-pressure liquid chromatographic UV method above 2 ppb of N, which is the detection limit of the latter method. It should be possible to analyze all types of samples for nitrate, except those with inhibiting substances, by this method. To illustrate the use of the denitrifier method, NO(3) concentrations of <2 ppb of NO(3) N were measured in distilled and deionized purified water samples and in anaerobic lake water samples, but were not detected at the surface of the sediment. The denitrifier method was also used to measure the atom% of N in NO(3). This method avoids the incomplete reduction and contamination of the NO(3) -N by the NH(4) and N(2) pools which can occur by the conventional method of NO(3) analysis. N(2)O-producing denitrifier strains were also used to measure the apparent K(m) values for NO(3) use by these organisms. Analysis of N(2)O production by use of a progress curve yielded K(m) values of 1.7 and 1.8 muM NO(3) for the two denitrifier strains studied. PMID- 16347652 TI - Extraction from natural planktonic microorganisms of DNA suitable for molecular biological studies. AB - We developed a simple technique for the high-yield extraction of purified DNA from mixed populations of natural planktonic marine microbes (primarily bacteria). This is a necessary step for several molecular biological approaches to the study of microbial communities in nature. The microorganisms from near shore marine and brackish water samples, ranging in volume from 8 to 40 liters, were collected on 0.22-mum-pore-size fluorocarbon-based filters, after prefiltration through glass fiber filters, to remove most of the eucaryotes. DNA was extracted directly from the filters in 1% sodium dodecyl sulfate that was heated to 95 to 100 degrees C for 1.5 to 2 min. This procedure lysed essentially all the bacteria and did not significantly denature the DNA. The DNA was purified by phenol extraction, and precautions were taken to minimize shearing. Agarose gel electrophoresis showed that most of the final preparation had a large molecular size (>23 kilobase pairs). The DNA was sufficiently pure to allow complete digestion by the restriction endonuclease Sau3AI and ligation to vector DNA. In a sample in which the extracted DNA was quantified by binding to the dye Hoechst H33258, DNA was quantitatively extracted, and 45% of the initially extracted DNA was recovered after purification. Final yields were a few micrograms of DNA per liter of seawater and were roughly 25 to 50% of the total bacterial DNA in the sample. Alternatives to the initial harvest by filtration method, including continuous-flow centrifugation and thin-channel or hollow-fiber concentration followed by centrifugation, were less efficient than filtration in terms of both time and yield, largely because of the difficulty of centrifuging the very small bacteria typical of marine plankton. These methods were judged to be less appropriate for studies of natural populations as they impose a strong selection for the larger bacteria. PMID- 16347653 TI - Sources of variability in the measurement of fungal spore yields. AB - VARIABILITY IN THE PRODUCTION OF FUNGAL SPORES AND IN THE MEASUREMENT OF SPORE YIELDS WAS INVESTIGATED IN FOUR SPECIES OF FUNGI: Colletotrichum gloeosporioides, Colletotrichum coccodes, Colletotrichum phomoides, and Acremonium strictum. When the fungi were grown on solid medium in microplates and spore yields were measured by counting the subsamples with a hemacytometer, the variability among hemacytometer squares was always the largest source of variation, accounting for 51 to 91% of the total variation. Variability among replicate cultures and results of repeat experiments were generally also significant. The effect of square-to-square variability on the precision of spore yield measurement was minimized by counting a moderate number (ca. 30) of squares per culture. Culture to-culture variability limited the practical precision of spore production measurements to a 95% confidence interval of approximately the mean +/- 25%. We provide guidelines for determining the number of replicate cultures required to attain this or other degrees of precision. Particle counter-derived spore counts and counts based on spore weights were much less variable than were hemacytometer counts, but they did not improve spore production estimates very much because of culture-to-culture variability. Results obtained by both of these methods differed from those obtained with a hemacytometer; particle counter measurements required a correction for spore pairs, while the relationship between spore weights and spore counts changed as the cultures aged. PMID- 16347654 TI - Bioluminescence Assay for Measuring the Number of Bacteria Adhering to the Hydrocarbon Phase in the BATH Test. AB - A thorough validation of the bacterial adherence to hydrocarbons (BATH) test was performed by means of a bioluminescence assay. Ten different gram-negative strains were subjected to the BATH test. For the calculation of the adhesion index, several factors had to be taken into account: ATP leakage, the action of ATP-hydrolyzing enzymes, the change in the extraction efficiency of Nucleotide Releasing Reagent for Microbial Cells (NRB; Lumac bv) after vortexing and the difference in light production after the addition of NRB. When the adhesion index values obtained by bioluminescence measurement were used as reference, the total plate count technique appeared to be unreliable in estimating the number of bacteria adhering to the hydrocarbon phase. A highly significant correlation was established, however, between those reference values and the adhesion index values obtained by the optical density reading for octane and especially for hexadecane. With xylene, no correlation was found between the optical density reading values and the total plate count or bioluminescence values. PMID- 16347655 TI - Microbially Mediated Mn(II) Oxidation in an Oligotrophic Arctic Lake. AB - Rates of oxidation of Mn(II) were measured by an in situ incubation technique in the water column of Toolik Lake, Alaska. Measured rates were lower than those observed in other aquatic systems but were sufficient to oxidize all Mn(II) in the lake within a 3-month period. Measured rates compared favorably with rates estimated from a previous study of the geochemical cycling of Mn in Toolik Lake. The Mn(II) oxidation was largely microbially mediated, as indicated by inhibition of oxidation rates by sodium azide. Azide had been previously demonstrated to be a suitable microbial poison for studying Mn(II) oxidation in seawater. This study demonstrates that azide is also a suitable poison for freshwaters and that it inhibits microbial but not abiotic oxidation of Mn(II). Manganese(II) oxidation rates were similar during cold, under-ice conditions in early spring and during warmer summer conditions. This observation suggests that Mn(II) concentration, rather than temperature or oxygen concentration, is the most important factor regulating Mn(II) oxidation rates in Toolik Lake. PMID- 16347656 TI - Hydrogen partial pressures in a thermophilic acetate-oxidizing methanogenic coculture. AB - Hydrogen partial pressures were measured in a thermophilic coculture comprised of a eubacterial rod which oxidized acetate to H(2) and CO(2) and a hydrogenotrophic methanogen, Methanobacterium sp. strain THF. Zinder and Koch (S. H. Zinder and M. Koch, Arch. Microbiol. 138:263-272, 1984) originally predicted, on the basis of calculations of Gibbs free energies of reactions, that the H(2) partial pressure near the midpoint of growth of the coculture should be near 4 Pa (ca. 4 x 10 atm; ca. 0.024 muM dissolved H(2)) for both organisms to be able to conserve energy for growth. H(2) partial pressures in the coculture were measured to be between 20 and 50 Pa (0.12 to 0.30 muM) during acetate utilization, approximately one order of magnitude higher than originally predicted. However, when DeltaG(f) (free energy of formation) values were corrected for 60 degrees C by using the relationship DeltaG(f) = DeltaH(f) - TDeltaS (DeltaH(f) is the enthalpy or heat of formation, DeltaS is the entropy value, and T is the temperature in kelvins), the predicted value was near 15 Pa, in closer agreement with the experimentally determined values. The coculture also oxidized ethanol to acetate, a more thermodynamically favorable reaction than oxidation of acetate to CO(2). During ethanol oxidation, the H(2) partial pressure reached values as high as 200 Pa. Acetate was not used until after the ethanol was consumed and the H(2) partial pressure decreased to 40 to 50 Pa. After acetate utilization, H(2) partial pressures fell to approximately 10 Pa and remained there, indicating a threshold for H(2) utilization by the methanogen. Axenic cultures of the acetate-oxidizing organism were combined with pure cultures of either Methanobacterium sp. strain THF or Methanobacterium thermoautotrophicum DeltaH to form reconstituted acetate oxidizing cocultures. The H(2) partial pressures measured in both of these reconstituted cocultures were similar to those measured in the original acetate oxidizing rod coculture. Since M. thermoautotrophicum DeltaH did not use formate as a substrate, formate is not necessarily involved in interspecies electron transfer in this coculture. PMID- 16347657 TI - Isolation and preliminary characterization of twenty bacteriophages infecting either brevibacterium or arthrobacter strains. AB - Thirty-seven bacteriophages plaquing on Corynebacterium, Brevibacterium, or Arthrobacter strains were isolated from soil or vegetation samples. Restriction analysis of phage DNA indicated that 20 phages were unique; one of them produced entirely turbid plaques on Brevibacterium ketoglutamicum and was characterized as temperate. All these phages were assigned to group B of the classification of Bradley (Bacteriol. Rev. 31:230-314, 1967) and had relatively narrow host ranges. PMID- 16347658 TI - Novel mode of microbial energy metabolism: organic carbon oxidation coupled to dissimilatory reduction of iron or manganese. AB - A dissimilatory Fe(III)- and Mn(IV)-reducing microorganism was isolated from freshwater sediments of the Potomac River, Maryland. The isolate, designated GS 15, grew in defined anaerobic medium with acetate as the sole electron donor and Fe(III), Mn(IV), or nitrate as the sole electron acceptor. GS-15 oxidized acetate to carbon dioxide with the concomitant reduction of amorphic Fe(III) oxide to magnetite (Fe(3)O(4)). When Fe(III) citrate replaced amorphic Fe(III) oxide as the electron acceptor, GS-15 grew faster and reduced all of the added Fe(III) to Fe(II). GS-15 reduced a natural amorphic Fe(III) oxide but did not significantly reduce highly crystalline Fe(III) forms. Fe(III) was reduced optimally at pH 6.7 to 7 and at 30 to 35 degrees C. Ethanol, butyrate, and propionate could also serve as electron donors for Fe(III) reduction. A variety of other organic compounds and hydrogen could not. MnO(2) was completely reduced to Mn(II), which precipitated as rhodochrosite (MnCO(3)). Nitrate was reduced to ammonia. Oxygen could not serve as an electron acceptor, and it inhibited growth with the other electron acceptors. This is the first demonstration that microorganisms can completely oxidize organic compounds with Fe(III) or Mn(IV) as the sole electron acceptor and that oxidation of organic matter coupled to dissimilatory Fe(III) or Mn(IV) reduction can yield energy for microbial growth. GS-15 provides a model for how enzymatically catalyzed reactions can be quantitatively significant mechanisms for the reduction of iron and manganese in anaerobic environments. PMID- 16347659 TI - Adaptation of phytoplankton-degrading microbial communities to thermal reactor effluent in a new cooling reservoir. AB - In water column and sediment inocula from a nuclear reactor cooling reservoir, natural phytoplankton substrate labeled with C was used to determine aerobic and anaerobic mineralization rates for a range of temperatures (25, 40, 55, and 70 degrees C) expected during reactor operation. For experiments that were begun during reactor shutdown, aerobic decomposition occurred at temperatures of <55 degrees C. After 2 months of reactor operation, aerobic rates increased substantially at 55 and 70 degrees C, although maximum rates were observed at temperatures of 0.99) was the highest (97 to 99%) with Vicia sativa, Trifolium repens, and Glycine max and lowest (72%) with Leucaena leucocephala. Medicago sativa and Macroptilium atropurpureum yielded 87 and 75% acceptable MPN codes, respectively. The acceptability of the MPN data obtained for a host species was related to rooting habit and time to nodulation. Comparison of data for each soil indicated that, despite large differences in characteristics, the soil was not a major source of variability in the MPN counts. There was no significant interaction of the range of transition of rhizobium-legume plant infection count data between species and site. PMID- 16347662 TI - Production and Characteristics of Raw-Potato-Starch-Digesting alpha-Amylase from Bacillus subtilis 65. AB - A newly isolated bacterium, identified as Bacillus subtilis 65, was found to produce raw-starch-digesting alpha-amylase. The electrophoretically homogeneous preparation of enzyme (molecular weight, 68,000) digested and solubilized raw corn starch to glucose and maltose with small amounts of maltooligosaccharides ranging from maltotriose to maltoheptaose. This enzyme was different from other amylases and could digest raw potato starch almost as fast as it could corn starch, but it showed no adsorbability onto any kind of raw starch at any pH. The mixed preparation with Endomycopsis glucoamylase synergistically digested raw potato starch to glucose at 30 degrees C. The raw-potato-starch-digesting alpha amylase showed strong digestibility to small substrates, which hydrolyzed maltotriose to maltose and glucose, and hydrolyzed p-nitrophenyl maltoside to p nitrophenol and maltose, which is different from the capability of bacterial liquefying alpha-amylase. PMID- 16347663 TI - Production and Characteristics of Avicel-Digesting and Non-Avicel-Digesting Cellobiohydrolases from Aspergillus ficum. AB - Two immunologically related cellobiohydrolases, cellobiohydrolase I (CBH I) and cellobiohydrolase II (CBH II), were purified from Aspergillus ficum. The Avicel adsorbable CBH I (molecular weight, 128,000) digested Avicel, cotton, and cellulose powder to cellobiose, but the Avicel-unadsorbable CBH II (molecular weight, 50,000) could not digest those substrates. Both enzymes hydrolyzed insoluble cellooligosaccharides (DP 25) to cellobiose. High-pressure liquid chromatographic analysis of soluble cellooligosaccharide hydrolysates revealed that both enzymes split off strictly cellobiose units from the nonreducing end of the cellulose chain with an exowise mechanism. CBH I showed glucosyltransferase activity, but CBH II did not. The N-bromosuccinimideoxidized CBH I was completely inactive but retained the ability to adsorb to Avicel. This suggested that CBH I has separate sites for binding to cellulose and for catalyzing cleavage of glycosidic linkages. Cellobiohydrolases were of two types, CBH I and CBH II. The former can adsorb to and digest Avicel, while the latter can do neither. PMID- 16347664 TI - An acidophilic and a neutrophilic nitrobacter strain isolated from the numerically predominant nitrite-oxidizing population of an Acid forest soil. AB - Two physiologically and serologically distinct strains of chemoautotrophic nitrite-oxidizing bacteria were isolated as numerically predominant members of the nitrite-oxidizer population of an undisturbed forest soil with a pH range of 4.3 to 5.2. One isolate responded as a neutrophile, characteristic of the family Nitrobacteraceae, and cross-reacted strongly with fluorescent antibody to Nitrobacter strain Engel. The second isolate responded as an acidophile in pure culture, demonstrated maximal nitrite oxidation activity at pH 5.5, and had a pH tolerance range of pH 4.1 to 7.2. Nitrite oxidase in whole cells of the acidophile sustained activity to at least pH 3.5. Cell morphology of both strains typified the genus Nitrobacter in all respects when cultured at pH 7. However, under more acidic conditions the acidophile tended to elongate and at times appeared to branch. These data provide the first evidence for the existence of an acidophilic chemoautotrophic nitrifying bacterium. Isolation of the neutrophilic Nitrobacter strain reported here complements the earlier isolation of a neutrophilic Nitrosospira strain to provide further evidence of a prominent acid intolerant population of chemoautotrophic nitrifiers in this acid forest soil. PMID- 16347665 TI - Cloning of Trametes versicolor Genes Induced by Nitrogen Starvation. AB - We have screened a genomic library of Trametes versicolor for genes whose expression is associated with nitrogen starvation, which has been shown to induce ligninolytic activity. Using two different approaches based on differential expression, we isolated 29 clones. These were shown by restriction mapping and cross-hybridization to code for 11 distinct differentially expressed genes. Northern analysis of the kinetics of expression of these genes revealed that at least four of them have kinetics of induction that parallel kinetics of induction of ligninolytic activity. PMID- 16347666 TI - Aerial Dispersal and Epiphytic Survival of Pseudomonas syringae during a Pretest for the Release of Genetically Engineered Strains into the Environment. AB - Prospective experimental field evaluation of genetically engineered microorganisms, such as microbial pest control agents, raises issues of how to properly ascertain their fate and survival in the environment. Field trials with recombinant organisms must reflect requirements for sampling and monitoring. Field trials were conducted at Tulelake, Calif., to monitor the numbers of viable cells of a nonrecombinant strain of Pseudomonas syringae that entered the atmosphere and landed on plants and soil during and after an aerosol spray application. An exponential decrease in numbers of viable cells deposited at increasing distances from three sprayed plots was observed. The relative rate of survival of cells sprayed directly on plants was more than 10 times higher than that of cells dispersed through the air to similar adjacent plants. Results are being used to gain experience with the characteristics of a release site that influence containment or dispersal and to develop appropriate sampling methodologies for evaluating survival and dispersal characteristics of genetically engineered bacteria released into the environment. The ability to make predictions about microbial dispersal and survival will reduce the uncertainties associated with environmental releases of recombinant organisms. PMID- 16347667 TI - Phospholipid Fatty Acid Composition of the Syntrophic Anaerobic Bacterium Syntrophomonas wolfei. AB - The membrane phospholipid fatty acids (PLFAs) from several cocultures and a pure culture of Syntrophomonas wolfei were determined by capillary column gas chromatography. Cocultures of S. wolfei with a Desulfovibrio sp. contained PLFAs from both organisms, whereas PLFAs from a coculture with Methanospirillum hungatei contained very little biomass to analyze. The pure culture of S. wolfei grown on crotonate provided the best material for analysis of the PLFAs. The predominant PLFAs of S. wolfei were the monounsaturated 16:1omega7c and 16:1omega9c and the saturated 16:0 and 14:0. A low concentration of the diunsaturated 18:2omega6 was detected. The PLFA analysis provides additional information for consideration in the determination of the profile of PLFAs obtained from anaerobic environments. In addition, this information may aid in the understanding of the physiology and phylogeny of S. wolfei and other syntrophic bacteria. PMID- 16347668 TI - l-Methionine, a Precursor of Trace Methane in Some Proteolytic Clostridia. AB - The in vivo formation of methane and of several S-methyl volatile compounds from the terminal S-methyl group of l-methionine is reported for growing cultures of four Clostridium strains (C. hastiforme, C. histolyticum, C. subterminale, and Clostridium sp. strain DSM 1786). After growth in 5 ml of unamended medium, C. hastiforme formed the highest amount of methane (408 nmol per tube in the headspace). When the culture medium was amended with 100 mM l-[S-methyl H(3)]methionine, the four strains formed [H(3)]methane (proportion in the methane peak, >85%) as well as methanethiol, dimethyl disulfide, dimethyl trisulfide, and S-methyl thioacetate labeled on the methyl moiety. Methanethiol is also a precursor of methane for Clostridium sp. strain DSM 1786. The trace methane formation observed for these four proteolytic, nonglucidolytic Clostridium strains can be of ecological interest, particularly in aquatic sediments and in the gastrointestinal tract of humans and animals. It can explain in part the trace methane formation which cannot be ascribed to methanogens sensu stricto. PMID- 16347669 TI - Genetic Transformation of an argB Mutant of Aspergillus oryzae. AB - An argB mutant of Aspergillus oryzae NRRL 492 has been genetically transformed with the Aspergillus nidulans argB gene. Protoplasts were generated with a combination of Novozyme 234 and beta-glucuronidase and regenerated on sucrose stabilized minimal medium without arginine as described for A. nidulans. A frequency of 5 to 10 transformants per mug of DNA was obtained; however, most transformants appeared abortive. The A. nidulans argB gene and vector sequences appeared to be integrated into the A. oryzae chromosome. PMID- 16347670 TI - Synthesis of Exopolysaccharide by Bradyrhizobium japonicum during Growth on Hydroaromatic Substrates. AB - The hydroaromatic acids shikimate and quinate, which may be available as carbon sources in the soil, supported production of only low levels of acidic exopolysaccharide by Bradyrhizobium japonicum. Exopolysaccharide production (micrograms per 10 cells) was 4.9 on quinate and 4.5 on shikimate; in comparison, it was 128 on adipate, 18 on l-arabinose, and 39 on d-glucose. PMID- 16347671 TI - A pH Control System Based on Malate Decarboxylation for the Cultivation of Lactic Acid Bacteria. AB - Most species of lactic acid bacteria decarboxylate l-malate to lactate and CO(2) if an energy source such as glucose is present. A proton is taken up in the reaction, which prevents pH decreases in the growth medium caused by lactic acid production from glucose fermentation. MRS broth (pH 7.0) (Difco Laboratories) containing 10 mM glucose and various concentrations of l-malate (0, 25, 50, 75, and 100 mM) was used to cultivate Lactobacillus plantarum. After 72 h at 37 degrees C, all malate was decarboxylated and all glucose was fermented, with resultant final pH values of 4.5, 6.3, 6.9, 7.3, and 7.5, respectively. When d malate (which cannot be decarboxylated) was substituted for l-malate, the final pH values were 4.5, 5.2, 5.6, 5.8, and 5.9. By varying the ratios of glucose to l malate in the growth medium, it was possible to obtain pH values which were lower, the same, or higher than the initial pH values. In contrast, buffers such as phosphate only retard decreases in pH. l-Malate, when compared with K(2)PO(4) on an equal molar basis, provided greater resistance to decreases in pH. Higher specific growth rates were observed for L. plantarum and Leuconostoc mesenteroides when l-malate rather than K(2)PO(4) was incorporated into the growth medium. PMID- 16347673 TI - Trichloroethylene metabolism by microorganisms that degrade aromatic compounds. AB - [This corrects the article on p. 605 in vol. 54.]. PMID- 16347672 TI - A simple, rapid method for enumerating total viable and metabolically active bacteria in groundwater. AB - We report here a new staining procedure which uses both the enzymatic dehydrogenation of 2-(p-iodophenyl)-3-p-(nitrophenyl)-5 phenyltetrazolium chloride to a pink intracellular formazan and the DNA-specific fluorochrome 4',6' diamidino-2-phenylindole. Application of this staining procedure to cells concentrated on filters and then transferred to microscope slides by the filter transfer-freeze technique has proven valuable for statistically accurate enumeration of the total viable and metabolically active cells in groundwaters. PMID- 16347674 TI - Crosses among Homokaryons from Commercial and Wild-Collected Strains of the Mushroom Agaricus brunnescens (= A. bisporus). AB - A new technique for the production of hybrid strains of the cultivated mushroom Agaricus brunnescens is described. Homokaryons were recovered from regenerated protoplasts obtained from several heterokaryotic strains. A total of 16 novel hybrids were produced in 63 attempted crosses between paired homokaryons. Recovery of both homokaryons and hybrids was verified by analysis of restriction fragment length polymorphisms. Three of four hybrids fruited in small-scale tests, further confirming that the isolates were true hybrids. Colony morphology alone was found to be a poor indicator of hybrid status. In two instances, three homokaryons crossed successfully in all combinations, suggesting that there are at least three alleles at the putative mating-type locus. Crosses between homokaryons from commercial and wild-collected isolates indicated that these strains belong to the same biological species. PMID- 16347675 TI - Diazotrophy and Nitrogenase Activity in the Archaebacterium Methanosarcina barkeri 227. AB - Nitrogen fixation (diazotrophy) has recently been demonstrated in several methanogenic archaebacteria. To compare the process in an archaebacterium with that in eubacteria, we examined the properties of diazotrophic growth and nitrogenase activity in Methanosarcina barkeri 227. Growth yields with methanol or acetate as a growth substrate were significantly lower in N(2)-grown cultures than in NH(4)-grown cultures, and the culture doubling times were increased, indicating that diazotrophy was energetically costly, as it is in eubacteria. Growth of nitrogen-fixing cells was inhibited when molybdenum was omitted from the medium; addition of 10 nM molybdate stimulated growth, while 1 muM molybdate restored maximum diazotrophic growth. Omission of molybdenum did not inhibit growth of ammonia-grown cells. Tungstate (100 muM) strongly inhibited growth of molybdenum-deficient diazotrophic cells, while ammonia-grown cells were unaffected. The addition of 100 nM vanadate or chromate did not stimulate diazotrophic growth of molybdenum-starved cells. These results are consistent with the presence of a molybdenum-containing nitrogenase in M. barkeri. Acetylene, the usual substrate for assaying nitrogenase activity, inhibited methanogenesis by M. barkeri and consequently needed to be used at a low partial pressure (0.3% of the headspace) when acetylene reduction by whole cells was assayed. Whole cells reduced 0.3% acetylene to ethylene at a very low rate (1 to 2 nmol h mg of protein), and they "switched off" acetylene reduction in response to added ammonia or glutamine. Crude extracts from diazotrophic cells reduced 10% acetylene at a rate of 4 to 5 nmol of C(2)H(4) formed h mg of protein when supplied with ATP and reducing power, while extracts of Klebsiella pneumoniae prepared by the same procedures had rates 100-fold higher. Acetylene reduction by extracts required ATP and was completely inhibited by 1 mM ADP in the presence of 5 mM ATP. The low rates of C(2)H(2) reduction could be due to improper assay conditions, to switched-off enzyme, or to the nitrogenase's having lower activity towards acetylene than towards dinitrogen. PMID- 16347676 TI - Enhancement of Butanol Formation by Clostridium acetobutylicum in the Presence of Decanol-Oleyl Alcohol Mixed Extractants. AB - Extractive fermentation has been proposed to enhance the productivity of fermentations that are end product inhibited. Unfortunately, good extractants for butanol, such as decanol, are toxic to Clostridium acetobutylicum. The use of mixed extractants, namely, mixtures of toxic and nontoxic coextractants, was proposed to circumvent this toxicity. Decanol appeared to inhibit butanol formation by C. acetobutylicum when present in a mixed extractant that also contained oleyl alcohol. However, maintenance of the pH at 4.5 alleviated the inhibition of butanol production and the consumption of butyrate during solventogenesis. A mixed extractant that contained 20% decanol in oleyl alcohol enhanced butanol formation by 72% under pH-controlled conditions. The production of acetone and acetoin was also increased, even though these two products were not extractable. The enhancement of butanol formation was not limited by the toxicity of decanol. Supplementation of glucose and butyrate in the extractive fermentation yielded a 47% increase in butanol. The enhancement of butanol formation appeared to be dependent on the presence of dissolved decanol in the broth but was not observed unless an organic phase was present to extract butanol. A mechanism for the effects of decanol on product formation is proposed. PMID- 16347677 TI - Chemotaxis in the Marine Fungus Rhizophydium littoreum. AB - Zoospores of the marine chytrid Rhizophydium littoreum are attracted to a variety of substances common to their environment. In general, carbohydrates and polysaccharides elicited strong concentration-dependent positive responses. There was no direct correlation between all substances used as foods and those stimulating positive responses. The chemotactic activities of this organism should, however, tend to bring it toward concentrated food sources. PMID- 16347678 TI - Ice nucleation activity in lichens. AB - A newly discovered form of biological ice nucleus associated with lichens is described. Ice nucleation spectra of a variety of lichens from the southwestern United States were measured by the drop-freezing method. Several epilithic lichen samples of the genera Rhizoplaca, Xanthoparmelia, and Xanthoria had nuclei active at temperatures as warm as -2.3 degrees C and had densities of 2.3 x 10 to more than 1 x 10 nuclei g at -5 degrees C (2 to 4 orders of magnitude higher than any plants infected with ice nucleation-active bacteria). Most lichens tested had nucleation activity above -8 degrees C. Lichen substrates (rocks, plants, and soil) showed negligible activity above -8 degrees C. Ice nucleation-active bacteria were not isolated from the lichens, and activity was not destroyed by heat (70 degrees C) or sonication, indicating that lichen-associated ice nuclei are nonbacterial in origin and differ chemically from previously described biological ice nuclei. An axenic culture of the lichen fungus Rhizoplaca chrysoleuca showed detectable ice nucleation activity at -1.9 degrees C and an ice nucleation density of 4.5 x 10 nuclei g at -5 degrees C. It is hypothesized that these lichens, which are both frost tolerant and dependent on atmospheric moisture, derive benefit in the form of increased moisture deposition as a result of ice nucleation. PMID- 16347679 TI - Mechanisms of DNA utilization by estuarine microbial populations. AB - The mechanisms of utilization of DNA by estuarine microbial populations were investigated by competition experiments and DNA uptake studies. Deoxyribonucleoside monophosphates, thymidine, thymine, and RNA all competed with the uptake of radioactivity from [H]DNA in 4-h incubations. In 15-min incubations, deoxyribonucleoside monophosphates had no effect or stimulated [H]DNA binding, depending on the concentration. The uptake of radioactivity from [H]DNA resulted in little accumulation of trichloroacetic acid-soluble intracellular radioactivity and was inhibited by the DNA synthesis inhibitor novobiocin. Molecular fractionation studies indicated that some radioactivity from [H]DNA appeared in the RNA (10 and 30% at 4 and 24 h, respectively) and protein (approximately 3%) fractions. The ability of estuarine microbial assemblages to transport gene sequences was investigated by plasmid uptake studies, followed by molecular probing. Although plasmid DNA was detected on filters after filtration of plasmid-amended incubations, DNase treatment of filters removed this DNA, indicating that there was little transport of intact gene sequences. These observations led to the following model for DNA utilization by estuarine microbial populations. (i) DNA is rapidly bound to the cell surface and (ii) hydrolyzed by cell-associated and extracellular nonspecific nucleases. (iii) DNA hydrolysis products are transported, and (iv) the products are rapidly salvaged into nucleic acids, with little accumulation into intracellular nucleotide pools. PMID- 16347680 TI - Effects of Glucose Concentrations on Cadmium, Copper, Mercury, and Zinc Toxicity to a Klebsiella sp. AB - The influence of glucose concentration on Cd, Cu, Hg, and Zn toxicity to a Klebsiella sp. was studied by following the degradation of C-labeled glucose at pH 6.0. Uptake of C into the cells was also determined. The carbon concentrations ranged from 0.01 to 40 mg liter, which are equivalent to soluble C concentrations in natural environments. The toxicity of Cu, Cd, and Zn to a Klebsiella sp. was affected considerably by the C concentration. Copper at 10 M was toxic when the carbon concentration was 10 or 40 mg liter, while at 0.01 to 1.0 mg liter no toxicity was observed. Cadmium and zinc were toxic at 10 M in media containing 0.01 to 1.0 mg of C liter. At C concentrations greater than 1.0 mg liter, the inhibition of glucose degradation and carbon assimilation was observed at 10 M Cd and Zn. The toxicity of mercury seemed to be independent of the C concentration. Results of this study showed that the nutritional state of an organism may have a profound effect on its sensitivity to metals. Metals taken up by an energy-driven transport system may be less toxic under conditions of C starvation. The C concentration should be taken into account when evaluating results from toxicity studies, especially as most microorganisms in nature live under energy-limited conditions. PMID- 16347681 TI - Oxidation of Ferrous Iron and Elemental Sulfur by Thiobacillus ferrooxidans. AB - The oxidation of ferrous iron and elemental sulfur by Thiobacillus ferrooxidans that was absorbed and unabsorbed onto the surface of sulfur prills was studied. Unadsorbed sulfur-grown cells oxidized ferrous iron at a rate that was 3 to 7 times slower than that of ferrous iron-grown cells, but sulfur-grown cells were able to reach the oxidation rate of the ferrous iron-adapted cells after only 1.5 generations in a medium containing ferrous iron. Bacteria that were adsorbed to sulfur prills oxidized ferrous iron at a rate similar to that of unadsorbed sulfur-grown bacteria. They also showed the enhancement of ferrous iron oxidation activity in the presence of ferrous iron, even though sulfur continued to be available to the bacteria in this case. An increase in the level of rusticyanin together with the enhancement of the ferrous iron oxidation rate were observed in both sulfur-adsorbed and unadsorbed cells. On the other hand, sulfur oxidation by the adsorbed bacteria was not affected by the presence of ferrous iron in the medium. When bacteria that were adsorbed to sulfur prills were grown at a higher pH (ca. 2.5) in the presence of ferrous iron, they rapidly lost both ferrous iron and sulfur oxidation capacities and became inactive, apparently because of the deposition of a jarosite-like precipitate onto the surface to which they were attached. PMID- 16347682 TI - Mineralization of trichloroethylene by heterotrophic enrichment cultures. AB - Microbial consortia capable of aerobically degrading more than 99% of exogenous trichloroethylene (TCE) (50 mg/liter) were collected from TCE-contaminated subsurface sediments and grown in enrichment cultures. TCE at concentrations greater than 300 mg/liter was not degraded, nor was TCE used by the consortia as a sole energy source. Energy sources which permitted growth included tryptone yeast extract, methanol, methane, and propane. The optimum temperature range for growth and subsequent TCE consumption was 22 to 37 degrees C, and the pH optimum was 7.0 to 8.1. Utilization of TCE occurred only after apparent microbial growth had ceased. The major end products recovered were hydrochloric acid and carbon dioxide. Minor products included dichloroethylene, vinylidine chloride, and, possibly, chloroform. PMID- 16347683 TI - Immobilization of leachable toxic soil pollutants by using oxidative enzymes. AB - Screening of leachable toxic chemicals in a horseradish peroxidase-H(2)O(2) immobilization system established that immobilization was promising for most phenolic pollutants but not for benzoic acid, 2,6-dinitrocresol, or dibutyl phthalate. The treatment did not mobilize inherently nonmobile pollutants such as anilines and benzo[a]pyrene. In a separate study, an extracellular laccase in the culture filtrate of Geotrichum candidum was selected from five fungal enzymes evaluated as a cost-effective substitute for horseradish peroxidase. This enzyme was used in demonstrating the immobilization and subsequent fate of C-labeled 4 methylphenol and 2,4-dichlorophenol in soil columns. When applied to Lakewood sand, 98.1% of 4-methylphenol was leached through with distilled water. Two days after immobilization treatment with the G. candidum culture filtrate, only 9.1% of the added 4-methylphenol was leached with the same volume of water. Of the more refractory test pollutant 2,4-dichlorophenol, 91.6% had leached at time zero and 48.5% had leached 1 day after the immobilization treatment. However, 2 weeks after immobilization, only 12.0% of the 2,4-dichlorophenol was leached compared with 61.7% from the control column that received no immobilization treatment. No remobilization of the bound pollutants was detected during 3- and 4-week incubation periods. Enzymatic immobilization of phenolic contaminants in soil appears to be a promising technique for the reduction of groundwater pollution by such substances. PMID- 16347684 TI - Photoexcretion and fate of glycolate in a hot spring cyanobacterial mat. AB - Photosynthesis by Synechococcus lividus, the sole oxygenic phototroph inhabiting the surface of the 55 degrees C cyanobacterial mat in Mushroom Spring, Yellowstone National Park, causes superoxic and alkaline conditions which promote glycolate photoexcretion. At O(2) concentrations characteristic of the top 2 mm of mat during the day, up to 11.8% of NaHCO(3) fixed in the light was excreted, and glycolate accounted for up to 58% of the excreted photosynthate. Glycolate was neither incorporated nor metabolized by S. lividus, but it was incorporated by filamentous microorganisms in the mat. Incubation of mat samples with NaHCO(3) resulted in labeling of both S. lividus and filaments, but the addition of nonradioactive glycolate increased the level of C in the aqueous phase and decreased the extent of labeling of filaments. This suggests that cross-feeding of glycolate from S. lividus to filamentous heterotrophs occurs and that underestimation of the extent of photoexcretion is probable. PMID- 16347685 TI - Molecular Cloning and Expression of Cellulase Genes from Ruminococcus albus 8 in Escherichia coli Bacteriophage lambda. AB - A genomic library of Ruminococcus albus 8 DNA was constructed by using the Escherichia coli bacteriophage lambdaDASH. Recombinants were screened for cellulolytic activity by plating in soft agar (0.7%) overlays containing either 1% (wt/vol) carboxymethyl cellulose (CMC), 4-methylumbelliferyl-beta-d cellobioside (MUC, 1 mg/ml), or 1% (wt/vol) Ostazin brilliant red-hydroxyethyl cellulose (OBR-HEC). One hundred and three recombinant phage exhibiting activity against OBR-HEC were found, and these fell into different classes based on the size of the zone of hydrolysis. Twenty-one recombinant phage exhibiting activity against CMC and 19 recombinant phage exhibiting activity against MUC were isolated. Four OBR-HEC, five CMC, and seven MUC clones were further analyzed by restriction endonuclease mapping and cellulase substrate specificity to identify unique clones and to determine their cellulase type. Three different clone types representing endoglucanase activity were identified. Three clones that appeared to encode exoglucanase type activity and four clones that had a mixed specificity, including beta-glucosidase activity, were also identified. PMID- 16347686 TI - Nitrile Hydratase-Catalyzed Production of Nicotinamide from 3-Cyanopyridine in Rhodococcus rhodochrous J1. AB - Nitrile hydratase, which occurs abundantly in cells of Rhodococcus rhodochrous J1 isolated from soil samples, catalyzes the hydration of 3-cyanopyridine to nicotinamide. By using resting cells, the reaction conditions for nicotinamide production were optimized. Under the optimum conditions, 100% of the added 12 M 3 cyanopyridine was converted to nicotinamide without the formation of nicotinic acid, and the highest yield achieved was 1,465 g of nicotinamide per liter of reaction mixture containing resting cells (1.48 g as dry cell weight) in 9 h. The nicotinamide produced was crystallized and then identified physicochemically. The further conversion of the nicotinamide to nicotinic acid was due to the low activity of nicotinamide as a substrate for the amidase(s) present in this organism. PMID- 16347687 TI - Conversion of Glucose to 2-Keto-l-Gulonate, an Intermediate in l-Ascorbate Synthesis, by a Recombinant Strain of Erwinia citreus. AB - A gene for 2,5-diketo-d-gluconate (25DKG) reductase, which encodes an enzyme composed of 277 amino acid residues catalyzing the reduction of 25DKG to 2-keto-l gulonate (2KLG), was cloned from Corynebacterium sp. strain SHS752001 and expressed in Erwinia citreus SHS2003, a strain which oxidizes glucose to 25DKG. The recombinant microorganism converted glucose to 2KLG, a compound which can be readily converted to l-ascorbate (vitamin C). Improvements in the yield of 2KLG were obtained by changing fermentation conditions, using the p(l) promoter of bacteriophage lambda to express the reductase, and selecting a mutant of E. citreus which could use neither 25DKG nor 2KLG as a sole carbon source for growth. When a culture of the recombinant strain was fed with glucose to a total of 40 g/liter, 49.4% of the glucose was converted to 2KLG during a 72-h fermentation. PMID- 16347688 TI - Production of mannitol by fungi from cotton dust. AB - Cotton dust associated with high pulmonary function decrements contains relatively high levels of mannitol. In this study, cotton leaf and bract tissue and dust isolated from cotton leaf tissue were examined by optical microscopy, scanning electron microscopy, and capillary gas chromatography. Alternaria alternata, Cladosporium herbarum, Epicoccum purpurascens, and Fusarium pallidoroseum were isolated from cotton leaf dust. The fungal samples, cotton dust, and cotton leaf contained mannitol. This study demonstrates that fungi from a late-fall harvest of cotton leaf material produce mannitol and are a probable source of the mannitol found in cotton dust. PMID- 16347689 TI - Nonspecific factors involved in attachment of entomopathogenic deuteromycetes to host insect cuticle. AB - The attachment of the conidia of the insect-pathogenic fungi Nomuraea rileyi, Beauveria bassiana, and Metarrhizium anisopliae to insect cuticle was mediated by strong binding forces. The attachment was passive and nonspecific in that the conidia adhered readily to both host and nonhost cuticle preparations. The hydrophobicity of the conidial wall and the insect epicuticle appeared to mediate the adhesion process. Detergents, solvents, and high-molecular-weight proteins known to neutralize hydrophobicity reduced conidial binding when added to conidium-cuticle preparations. However, these chemicals did not remove the hydrophobic components from the epicuticle or from conidial preparations. The outer surface of the conidium consists of a resilient layer of well-organized fascicles of rodlets. Intact rodlets extracted from B. bassiana conidia bound to insect cuticle and exhibited the hydrophobicity expressed by intact conidia. Both electrostatic charges and various hemagglutinin activities were also present on the conidial surface. However, competitive-inhibition studies indicated that these forces played little, if any, role in the adhesion process. PMID- 16347690 TI - Melanin production by Rhizobium strains. AB - Different Rhizobium and Bradyrhizobium strains were screened for their ability to produce melanin. Pigment producers (Mel) were found among strains of R. leguminosarum biovars viceae, trifolii, and phaseoli, R. meliloti, and R. fredii; none of 19 Bradyrhizobium strains examined gave a positive response. Melanin production and nod genes were plasmid borne in R. leguminosarum biovar trifolii RS24. In R. leguminosarum biovar phaseoli CFN42 and R. meliloti GR015, mel genes were located in the respective symbiotic plasmids. In R. fredii USDA 205, melanin production correlated with the presence of its smallest indigenous plasmid. PMID- 16347691 TI - O-Methylation of Chlorinated para-Hydroquinones by Rhodococcus chlorophenolicus. AB - Rhodococcus chlorophenolicus PCP-I, a degrader of polychlorinated phenols, guaiacols (2-methoxyphenols), and syringols (2,6-dimethoxyphenols), was shown to O-methylate the degradation intermediate, a chlorinated para-hydroquinone, into 4 methoxyphenol. O-methylation was constitutively expressed, whereas the degradation of chlorophenols and chlorohydroquinones was inducible in R. chlorophenolicus. The O-methylating reaction required two hydroxyl groups in positions para to each other. R. chlorophenolicus selectively methylated the hydroxyl group flanked by two chlorine substituents. Tetrachlorohydroquinone, trichlorohydroquinone, and 2,6-dichlorohydroquinone were methylated into tetrachloro-4-methoxyphenol, 2,3,5-trichloro-4-methoxyphenol, and 3,5-dichloro-4 methoxyphenol, respectively. Chlorohydroquinones with only one chlorine adjacent to a hydroxyl group were methylated only in trace amounts, and no metabolite was formed from hydroquinone. The degradation intermediates formed in hydroxylation of tetrachloroguaiacol and trichlorosyringol by R. chlorophenolicus were O methylated into two isomeric trichlorodimethoxyphenols and two isomeric dichlorotrimethoxyphenols, respectively. R. chlorophenolicus also degraded the polychlorinated methylation products (tetrachlorinated and trichlorinated 4 methoxyphenols), but not mono- and dichlorinated 4-methoxyphenols. PMID- 16347692 TI - Numerical Taxonomic Analysis of Some Strains of Rhizobium spp. That Uses a Qualitative Coding of Immunodiffusion Reactions. AB - Antigenic relationships among seven strains of Bradyrhizobium japonicum were examined by immunodiffusion reactions, in which cells of each strain were reacted against each of the seven corresponding antisera. Similar analyses were performed with Rhizobium trifolii (28 strains), Rhizobium meliloti (9 strains), and rhizobia of the cowpea miscellany (13 strains). Antigens and antisera were reacted within each species only; serological interspecies cross-reactions were not performed. The results, scored qualitatively as reactions of identity, cross reactions, or no reaction, were formed into datum matrices and used to analyze the relationships between strains by applying the association measure of Bray and Curtis (J. R. Bray and J. T. Curtis, Ecol. Monogr. 27:325-349, 1957) and the UPGMA clustering algorithm (P. H. A. Sneath and R. R. Sokal, Numerical Taxonomy, 1973). No two strains were regarded as being serologically identical unless each gave the same results as the other in each immunodiffusion reaction against every antiserum. Despite the high level of cross-reactions and reactions of identity (totalling 93% of all cell-antiserum combinations) among strains of R. trifolii and R. meliloti, no strains were identical by the criterion described above; however, the strains of these species clustered rapidly and fused at the 70% similarity level. The B. japonicum strains and the rhizobia of the cowpea miscellany were much less cross-reactive (67 and 86% of all combinations were negative, respectively), and they clustered more slowly. The strains of B. japonicum fused completely only at the 4% similarity level, whereas of the 13 cowpea-nodulating strains, 4 reacted as two pairs of identical strains and 6 remained unfused. PMID- 16347693 TI - Fluorescein Isothiocyanate-Labeled Lectin Analysis of the Surface of the Nitrogen Fixing Bacterium Azospirillum brasilense by Flow Cytometry. AB - The cell surface of Azospirillum brasilense was probed by using fluorescein isothiocyanate (FITC)-labeled lectins, with binding determined by fluorescence activated flow cytometry. Cells from nitrogen-fixing or ammonium-assimilating cultures reacted similarly to FITC-labeled lectins, with lectin binding in the following order: Griffonia simplicifolia II agglutinin > Griffonia simplicifolia I agglutinin > Triticum vulgaris agglutinin > Glycine max agglutinin > Canavalia ensiformis agglutinin > Limax flavus agglutinin > Lotus tetragonolobus agglutinin. The fluorescence intensity of cells labeled with FITC-labeled G. simplicifolia I, C. ensiformis, T. vulgaris, and G. max agglutinins was influenced by lectin concentration. Flow cytometry measurements of lectin binding to cells was consistent with measurements of agglutination resulting from lectin cell interaction. Capsules surrounding nitrogen-fixing and ammonium-assimilating cells were readily demonstrated by light and transmission electron microscopies. PMID- 16347694 TI - Effect of root agglutinin on microbial activities in the rhizosphere. AB - A total of 220 bacterial isolates were obtained from pea rhizosphere and nonrhizosphere samples. Of these samples, 100 isolates were chosen randomly to test for their agglutinative reaction against pea root exudate. The percentage of positive agglutination of bacteria isolated from the nonrhizosphere sample was significantly lower than that of bacteria isolated from the rhizosphere sample. Moreover, this agglutinative reaction could not be blocked either by treating the bacterial cells or root exudate with different carbohydrates before they were mixed or by boiling the root exudate first. Bacteria that could be agglutinated by pea root exudate followed the downward growth of the pea root through the soil profile. The greater abilities of such bacteria to colonize the pea rhizosphere were indicated by their higher rhizosphere-colonizing (rhizosphere/nonrhizosphere) ratios, whether the bacteria were added alone or together with nonagglutinating bacteria. However, bacteria did show different agglutinative reactions toward root exudates obtained from different plants. PMID- 16347695 TI - Effect of Temperature and Prey Availability on Growth of Paramoeba invadens in Monoxenic Culture. AB - Paramoeba invadens Jones 1985 is a pathogenic marine amoeba responsible for mass mortalities of sea urchins (Strongylocentrotus droebachiensis) of Nova Scotia between 1980 and 1983. A direct relationship between temperature and sea urchin paramoebiasis has been shown in previous laboratory and field studies. This study examined the effect of prey availability and temperature on the growth of P. invadens in monoxenic culture (with the marine bacterium Pseudomonas nautica). At 15 degrees C, the specific growth rate of P. invadens increased with bacterial prey concentration and was highest at 10 bacterial cells ml. Growth rate of P. invadens was maximal at 15 to 20 degrees C (which corresponds to annual sea temperature maxima in the natural environment) and the minimum generation time was 19.41 h at 20 degrees C. At 10 and 12 degrees C, generation times were 91.18 and 73.39 h, respectively; at 2 and 5 degrees C, there was no growth. P. invadens did not survive in monoxenic culture at 27 degrees C. Growth rates of P. invadens in vitro were positively correlated with time to morbidity of infected S. droebachiensis. PMID- 16347696 TI - Isolation of Plasmid-Harboring Serratia plymuthica from Facultative Gut Microflora of the Tobacco Hornworm, Manduca sexta. AB - Aseptic isolation of the facultative gut microflora of the tobacco hornworm, Manduca sexta, yielded four microorganisms. Two were gram-positive Bacillus spp., one was Serratia plymuthica, and another was the yeast Candida guilliermondii. The three bacterial species were screened for extrachromosomal DNA, and S. plymuthica was found to have a 6.4-kilobase plasmid, which was designated pCP-1. PMID- 16347697 TI - Oxidation of Thianthrene by the Ligninase of Phanerochaete chrysosporium. AB - The oxidation of heterocyclic sulfur compounds reported to be part of the macrostructure of coal and petroleum was investigated. The oxidation of thianthrene solubilized in 10% dimethylformamide to thianthrene monosulfoxide in the presence of hydrogen peroxide was catalyzed by the ligninase from Phanerochaete chrysosporium. PMID- 16347698 TI - Consequences of accounting for isotopic dilution in thymidine incorporation assays. AB - Rates of thymidine incorporation into DNA were corrected for isotope dilution by internal nucleotide pools and were compared with rates obtained from uncorrected data. Differences as large as 109% were observed between corrected and uncorrected estimates of thymidine incorporation. The degree of underestimation varied seasonally and, to a lesser extent, spatially. PMID- 16347699 TI - Acetate inhibition of methanogenic, syntrophic benzoate degradation. AB - Acetate inhibited benzoate degradation by a syntrophic coculture of an anaerobic benzoate degrader (strain BZ-2) and Methanospirillum strain PM-1; the apparent K(i) for acetate was approximately 40 mM. The addition of acetate resulted in a decrease in the hydrogen concentration in the coculture, indicating that phenomena related to interspecies hydrogen transfer affected this value and that the effect of acetate on the benzoate-degrading partner was probably greater than the apparent K(i) for the coculture suggests. PMID- 16347700 TI - Fundamental procedures for determining ergosterol content of decaying plant material by liquid chromatography. AB - Portions of published procedures for measurement of ergosterol content of decomposing plants were examined for their influence upon ergosterol yield. Common methods of treatment of plant samples prior to sterol extraction (e.g., oven drying, freezing, lyophilization) led to reduced recoveries of ergosterol (ca. 20 to 80%). The least destructive method was direct placement and storage in methanol. Photoconversion of ergosterol is not likely to cause losses during analysis, but losses are likely if there is insufficient mixing during neutral lipid partitioning from base-hydrolysis reagents. Homogenization (two times for 2 min) and refluxing (2 h) in methanol were equally effective in extracting ergosterol. Direct extraction in base-hydrolysis reagents was less effective (by ca. 40%). PMID- 16347701 TI - Phenolic Compound Utilization by the Soft Rot Fungus Lecythophora hoffmannii. AB - Nine phenolic compounds were metabolized by the soft rot fungus Lecythophora hoffmannii via protocatechuic acid and subsequently cleaved by protocatechuate 3,4-dioxygenase as determined by oxygen uptake, substrate depletion, and ring cleavage analysis. Catechol was metabolized by catechol 1,2-dioxygenase. Fungal utilization of these aromatic compounds may be important in the metabolism of wood decay products. PMID- 16347703 TI - Some Changes in Gut Bacterial Flora of Field-Grown Peridroma saucia (Lepidoptera: Noctuidae) When Brought into the Laboratory. AB - Removal of Peridroma saucia from the field to the laboratory caused little change in the quantity of facultative and aerobic bacteria in the gut but produced significant qualitative and quantitative changes in distinguishable groups of the family Enterobacteriaceae in the gut. PMID- 16347702 TI - Polysaccharide Production by Propionibacteria during Lactose Fermentation. AB - The fermentation products from 10 strains of propionibacteria accounted for only 72% (average value) of the lactose carbon utilized. The balance of the carbon was accounted for by the production of a polysaccharide containing methylpentose (the major component), glucose, and galactose. The presence of methylpentose explained the low ratios of propionate to acetate (<2:1). PMID- 16347704 TI - Restriction Fragment Length Polymorphisms in the Mushrooms Agaricus brunnescens and Agaricus bitorquis. AB - [This corrects the article on p. 817 in vol. 53.]. PMID- 16347705 TI - Nodulation and Nitrogen Fixation Efficacy of Rhizobium fredii with Phaseolus vulgaris Genotypes. AB - Phaseolus plant introduction (PI) genotypes (consisting of 684 P. vulgaris, 26 P. acutifolius, 39 P. lunatus, and 5 P. coccineus accessions) were evaluated for their ability to form effective symbioses with strains of six slow-growing (Bradyrhizobium) and four fast-growing (Rhizobium fredii) soybean rhizobia. Of the 684 P. vulgaris genotypes examined, three PIs were found to form effective nitrogen-fixing symbioses with the R. fredii strains. While none of the Bradyrhizobium strains nodulated any of the genotypes tested, some produced large numbers of undifferentiated root proliferations (hypertrophies). A symbiotic plasmid-cured R. fredii strain failed to nodulate the P. vulgaris PIs and cultivars, suggesting that P. vulgaris host range genes are Sym plasmid borne in the fast-growing soybean rhizobia. PMID- 16347706 TI - Estimating bacterial production in marine waters from the simultaneous incorporation of thymidine and leucine. AB - We examined the simultaneous incorporation of [H]thymidine and [C]leucine to obtain two independent indices of bacterial production (DNA and protein syntheses) in a single incubation. Incorporation rates of leucine estimated by the dual-label method were generally higher than those obtained by the single label method, but the differences were small (dual/single = 1.1 +/- 0.2 [mean +/- standard deviation]) and were probably due to the presence of labeled leucyl-tRNA in the cold trichloroacetic acid-insoluble fraction. There were no significant differences in thymidine incorporation between dual- and single-label incubations (dual/ single = 1.03 +/- 0.13). Addition of the two substrates in relatively large amounts (25 nM) did not apparently increase bacterial activity during short incubations (<5 h). With the dual-label method we found that thymidine and leucine incorporation rates covaried over depth profiles of the Chesapeake Bay. Estimates of bacterial production based on thymidine and leucine differed by less than 25%. Although the need for appropriate conversion factors has not been eliminated, the dual-label approach can be used to examine the variation in bacterial production while ensuring that the observed variation in incorporation rates is due to real changes in bacterial production rather than changes in conversion factors or introduction of other artifacts. PMID- 16347707 TI - Concomitant conjugal transfer of reduced-bacteriophage-sensitivity mechanisms with lactose- and sucrose-fermenting ability in lactic streptococci. AB - Ten previously reported lactose-positive (Lac) transconjugants from Streptococcus lactis, S. cremoris, and S. lactis subsp. diacetylactis and one sucrose-positive (Suc) transconjugant from S. lactis were examined for their sensitivity to prolate- and small isometric-headed bacteriophages. Four of the Lac transconjugants showed a 10- to 100-fold reduction in the efficiency of plating (EOP) as well as a reduced plaque size for the prolate phage c2 and were insensitive to the small isometric phage 712. A fifth Lac transconjugant demonstrated a similar reduced sensitivity to phage c2; however, this transconjugant was able to plaque phage 712, but with a reduced plaque size and EOP. The other five Lac transconjugants were sensitive to both c2 and 712 phages. The Suc transconjugant plaqued phage 712 with a reduced plaque size and EOP, but no reduction in plaque size or EOP was observed for phage c2. The Lac and reduced bacteriophage sensitivity (Rbs) phenotypes were correlated with specific plasmids in the Lac transconjugants. As four of the Lac transconjugants exhibited a phenotypically indistinguishable Rbs, one (AB001) was selected for further study. The Rbs in AB001 for both small isometric- and prolate-headed phages was not related to adsorption, and the reduced EOP for phage c2 was not related to the presence of a restriction and modification system. The latent period for phage c2 was unchanged, but the burst size was reduced 80%. The presence of the plasmid coding for Rbs retarded the lysis of a mitomycin C-induced prophage-containing strain. The Rbs mechanism appears to be abortive phage infection. This study supports previous observations that Rbs and conjugal transfer ability are physically linked among some group N streptococci. The results presented have implications in the identification of plasmids coding for Rbs and may also aid in explaining the dissemination of Rbs genes among lactic streptococci. PMID- 16347708 TI - Pseudomonas oleovorans as a Source of Poly(beta-Hydroxyalkanoates) for Potential Applications as Biodegradable Polyesters. AB - Pseudomonas oleovorans was grown in homogeneous media containing n-alkanoic acids, from formate to decanoate, as the sole carbon sources. Formation of intracellular poly(beta-hydroxyalkanoates) was observed only for hexanoate and the higher n-alkanoic acids. The maximum isolated polymer yields were approximately 30% of the cellular dry weight with growth on either octanoate or nonanoate. In most cases, the major repeating unit in the polymer had the same chain length as the n-alkanoic acid used for growth, but units with two carbon atoms less or more than the acid used as a carbon source were also generally present in the polyesters formed. Indeed, copolymers containing as many as six different types of beta-hydroxyalkanoate units were formed. The weight average molecular weights of the poly(beta-hydroxyalkanoate) copolymers produced by P. oleovorans ranged from 90,000 to 370,000. In spite of the higher cell yields obtained with octanoate and nonanoate, the use of hexanoate and heptanoate yielded higher-molecular-weight polymers. These copolyesters represent an entirely new class of biodegradable thermoplastics. PMID- 16347709 TI - Stress- and Growth Phase-Associated Proteins of Clostridium acetobutylicum. AB - The response of Clostridium acetobutylicum ATCC 4259 to the stresses produced by a temperature upshift from 28 degrees C to 45 degrees C and by exposure of the organisms to 0.1% n-butanol or to air was examined by analysis of pulse-labeled proteins. The stress response was the induction of the synthesis of a number of proteins, some of which were elicited by the three forms of stress. Eleven heat shock proteins were identified by two-dimensional electrophoresis, as were two proteins whose synthesis was heat sensitive. In the absence of applied stress, the synthesis of four proteins was found to be associated with the growth phase in batch culture; three of these proteins had a higher rate of de novo synthesis when the cells entered the solvent production phase. One of the stress-induced proteins, hsp74, was partially purified an found to be immunologically related to Escherichia coli heat shock protein Dnak. The similarities of the proteins induced at the onset of solventogenesis and by stress suggest a relationship between the two processes. PMID- 16347710 TI - Early Infection and Competition for Nodulation of Soybean by Bradyrhizobium japonicum 123 and 138. AB - Interactions of soybean with Bradyrhizobium japonicum 123 (serogroup 123) and 138 (serogroup c1) were used to examine the relationship between early infection rates, competition for nodulation, and patterns of nodule occupancy. Both strains formed more infections in autoclaved soil (sterile soil) than in untreated soil (unsterile soil). Inoculation did not increase numbers of infection threads in unsterile soil-grown plants, where infection of proximal portions of primary roots was complete by 5 days after planting. Both strains infected and nodulated at similar rates in sterile soil. Nodules were always clustered on the upper root system, regardless of inoculation and soil treatment. Sixty-seven percent of the nodules of uninoculated plants grown in unsterile soil were occupied by rhizobia belonging to serogroups other than 123 or c1. Inoculation with strain 123 or 138 increased occupancy by that strain at the expense of residency by other rhizobia. Eighty-three percent of all nodules on plants dually inoculated with both strains in sterile soil contained strain 138. The corresponding value for plants inoculated in unsterile soil was 31%. Neither inoculum strain dominated occupancy of first-formed nodules in unsterile soil. It appears that north central Missouri soil may not have populations of highly competitive serogroup 123 and that early infection and nodulation rates do not contribute to the competitive success of strain 138. PMID- 16347711 TI - Monoclonal Antibody Analysis and Insecticidal Spectrum of Three Types of Lepidopteran-Specific Insecticidal Crystal Proteins of Bacillus thuringiensis. AB - We have investigated the protein composition and the insecticidal spectrum of crystals of 29 Bacillus thuringiensis strains active against lepidopteran larvae. All crystals contained proteins of 130 to 140 kilodaltons (kDa) which could be grouped into three types by the molecular weight of the protoxin and the trypsin activated core fragment. Proteins of the three types showed a characteristic insecticidal spectrum when tested against five lepidopteran species. Type A crystal proteins were protoxins of 130 or 133 kDa, which were processed into 60 kDa toxins by trypsin. Several genes encoding crystal proteins of this type have been cloned and sequenced earlier. They are highly conserved in the N-terminal half of the toxic fragment and were previously classified in three subtypes (the 4.5-, 5.3-, and 6.6-kilobase subtypes) based on the restriction map of their genes. The present study shows that different proteins of these three subtypes were equally toxic against Manduca sexta and Pieris brassicae and had no detectable activity against Spodoptera littoralis. However, the 4.5-, 5.3-, and 6.6-kilobase subtypes differed in their toxicity against Heliothis virescens and Mamestra brassicae. Type B crystal proteins consisted of 140-kDa protoxins with a 55-kDa tryptic core fragment. These were only active against one of the five insect species tested (P. brassicae). The protoxin and the trypsin-activated toxin of type C were 135- and 63-kDa proteins, respectively. Proteins of this type were associated with high toxicity against S. littoralis and M. brassicae. A panel of 35 monoclonal antibodies was used to compare the structural characteristics of crystal proteins of the three different types and subtypes. Each type of protein could be associated with a typical epitope structure, indicating an unambiguous correlation between antigenic structure and insect specificity. PMID- 16347713 TI - Fusarium Wilt Suppression and Agglutinability of Pseudomonas putida. AB - Mutants of Pseudomonas putida (Agg) that lack the ability to agglutinate with components present in washes of bean and cucumber roots showed limited potential to protect cucumber plants against Fusarium oxysporum f. sp. cucumerinum. However, a higher level of protection was observed against Fusarium wilt in cucumber plants coinoculated with the parental bacterium (Agg), which was agglutinable. The Agg mutants did not colonize the roots of cucumber plants as extensively as the Agg parental isolate did. In competition experiments involving bean roots inoculated with a mixture of Agg and Agg bacteria, the Agg strains colonized roots to a greater extent than the Agg cells did. These data suggest that the Agg phenotype provides additional interactions that aid in the beneficial character of P. putida. PMID- 16347712 TI - Experimental evaluation of conversion factors for the [h]thymidine incorporation assay of bacterial secondary productivity. AB - The relationship between bacterial growth and incorporation of [methyl H]thymidine in oligotrophic lake water cultures was investigated. Prescreening, dilution, and addition of organic and inorganic nutrients were treatments used to prevent bacterivory and stimulate bacterial growth. Growth in unmanipulated samples was estimated through separate measurements of grazing losses. Both bacterial number and biovolume growth responses were measured, and incorporation of [H]thymidine in both total macromolecules and nucleic acids was assayed. The treatments had significant effects on conversion factors used to relate thymidine incorporation to bacterial growth. Cell number-based factors ranged from 1.1 x 10 to 38 x 10 cells mol of total thymidine incorporation and varied with treatment up to 10-fold for the same initial bacterial assemblage. In contrast, cell biovolume-based conversion factors were similar for two treatment groups across a 16-fold range of [H]thymidine incorporation rates: 5.54 x 10 mum mol of total thymidine incorporation and 15.2 x 10 mum mol of nucleic acid incorporation. Much of the variation in cell number-based conversion factors was related to changes in apparent mean cell volume of produced bacteria. Phosphorus addition stimulated [H]thymidine incorporation more than it increased bacterial growth, which resulted in low conversion factors. PMID- 16347714 TI - Properties of Thermus ruber Strains Isolated from Icelandic Hot Springs and DNA:DNA Homology of Thermus ruber and Thermus aquaticus. AB - Seventeen pink-pigmented strains of the genus Thermus were isolated from samples collected from thermal areas of Iceland. The strains were examined by using phenotypic characterization and DNA:DNA homology and were compared with recognized strains. Visually, the strains could be divided into three groups based on their pigmentation; however, spectroscopic studies of the pigments indicated little difference among them. Most strains required a vitamin supplement for growth and used fructose, maltose, mannose, or sucrose as the sole carbon source. In the presence of nitrate, two strains were able to grow under anaerobic conditions. The optimum growth temperature was 60 degrees C; growth did not occur at 30 or 70 degrees C. PMID- 16347715 TI - Kinetics of mixed microbial assemblages enhance removal of highly dilute organic substrates. AB - Our experiments with selected organic substrates reveal that the rate-limiting process governing microbial degradation rates changes with substrate concentration, S, in such a manner that substrate removal is enhanced at lower values of S. This enhancement is the result of the dominance of very efficient systems for substrate removal at low substrate concentrations. The variability of dominant kinetic parameters over a range of S causes the kinetics of complex assemblages to be profoundly dissimilar to those of systems possessing a single set of kinetic parameters; these findings necessitate taking a new approach to predicting substrate removal rates over wide ranges of S. PMID- 16347716 TI - Identification and characteristics of actinomycetes useful for semicontinuous treatment of domestic animal feces. AB - Selected strains of actinomycetes useful for practicing semicontinuous treatment of swine and poultry feces were identified as Streptomyces antibioticus S-4, S. puniceus N-50-2, S. nigrifaciens N-9-3, Thermoactinomyces vulgaris HIR-60, and Thermomonospora viridis HIR-50. These five obligately aerobic strains grew preferably on nonsterilized fresh swine feces in 24 h without any additives. They assimilated offensive volatile fatty acids in the swine and poultry feces. Cultures of these five strains were mixed and used as seed for the practical treatments of 1 ton of swine feces over the wide temperature range of 15 to 60 degrees C. Strain HIR-50 grew most predominantly on both fresh swine and poultry feces at 50 to 55 degrees C and decomposed uric acid. For the efficient penetration of mycelia into the feces, manures were mixed once a day so as not to break the solid mass, and the dehydration rate of feces had to be controlled in proportion to the mycelial growth rate. The actinomycete biofertilizer thus manufactured in 10 days was odorless and promotive of plant growth. PMID- 16347717 TI - Control of the Life Cycle of Methanosarcina mazei S-6 by Manipulation of Growth Conditions. AB - The morphology of Methanosarcina mazei was controlled by magnesium, calcium, and substrate concentrations and by inoculum size; these factors allowed manipulation of the morphology and interconversions between pseudosarcinal aggregates and individual, coccoid cells. M. mazei grew as aggregates in medium with a low concentration of catabolic substrate (either 50 mM acetate, 50 mM methanol, or 10 mM trimethylamine) unless Ca and Mg concentrations were high. Growth in medium high in Ca, Mg, and substrate (i.e., 150 mM acetate, 150 mM methanol, or 40 mM trimethylamine) converted pseudosarcinal aggregates to individual cocci. In such media, aggregates separated into individual cells which continued to grow exclusively as single cells during subsequent transfers. Conversion of single cells back to aggregates was complicated, because conditions which supported the aggregated morphology (e.g., low calcium or magnesium concentration) caused lysis of coccoid inocula. We recovered aggregates from coccoid cells by inoculating serial dilutions into medium high in calcium and magnesium. Cells from very dilute inocula grew into aggregates which disaggregated on continued incubation. However, timely transfer of the aggregates to medium low in calcium, magnesium, and catabolic substrates allowed continued growth as aggregates. We demonstrated the activity of the enzyme (disaggregatase) which caused the dispersion of aggregates into individual cells; disaggregatase was produced not only during disaggregation but also in growing cultures of single cells. Uronic acids, the monomeric constituents of the Methanosarcina matrix, were also produced during disaggregation and during growth as coccoids. PMID- 16347718 TI - Characterization of the adhesive holdfast of marine and freshwater caulobacters. AB - Caulobacters are prosthecate (stalked) bacteria that elaborate an attachment organelle called a holdfast at the tip of the cellular stalk. We examined the binding of lectins to the holdfasts of 16 marine Caulobacter strains and 10 freshwater species or strains by using a panel of fluorescein-conjugated lectins and fluorescence microscopy. The holdfasts of all the marine isolates bound to only wheat germ agglutinin (WGA) and other lectins that bind N-acetylglucosamine (GlcNac) residues. The freshwater caulobacters showed more variability in holdfast composition. Some bound only to WGA and comparable lectins as the marine strains did. Others bound additional or other lectins, and some did not bind to the lectins tested. The binding of WGA appeared to involve the regions of the holdfast involved with adhesion; a holdfast bound to WGA was significantly less adhesive to glass. Competition experiments with WGA-binding holdfasts and oligomers of GlcNac demonstrated that trimers of GlcNac (the preferred substrate for WGA binding) were more effective than dimers or monomers in preventing WGA binding to holdfasts, suggesting that stretches of contiguous GlcNac residues occur in the WGA-binding holdfasts. In addition, differences between freshwater and marine holdfasts in the strength of WGA binding were noted. The effect of a number of proteolytic and glycolytic enzymes on holdfast integrity was examined; the proteases had no effect for all caulobacters. None of the glycolytic enzymes had an effect on marine caulobacter holdfasts, but chitinase and lysozyme (both attack oligomers of GlcNac) disrupted the holdfasts of those freshwater caulobacters that bound WGA. Despite some similarity to chitin, holdfasts did not bind Calcofluor and no measurable effects on holdfast production were detectable after cell growth in the presence of diflubenzuron or polyoxin D, inhibitors of chitin synthesis in other systems. Finally, the holdfasts of all caulobacters bound to colloidal gold particles, without regard to the coating used to stabilize the gold particles. This binding was stronger or more specific than WGA binding; treatment with colloidal gold particles prevented WGA binding, but the reverse was not the case. PMID- 16347719 TI - Hydrogen peroxide metabolism in yeasts. AB - A catalase-negative mutant of the yeast Hansenula polymorpha consumed methanol in the presence of glucose when the organism was grown in carbon-limited chemostat cultures. The organism was apparently able to decompose the H(2)O(2) generated in the oxidation of methanol by alcohol oxidase. Not only H(2)O(2) generated intracellularly but also H(2)O(2) added extracellularly was effectively destroyed by the catalase-negative mutant. From the rate of H(2)O(2) consumption during growth in chemostat cultures on mixtures of glucose and H(2)O(2), it appeared that the mutant was capable of decomposing H(2)O(2) at a rate as high as 8 mmol . g of cells . h. Glutathione peroxidase (EC 1.11.1.9) was absent under all growth conditions. However, cytochrome c peroxidase (CCP; EC 1.11.1.5) increased to very high levels in cells which decomposed H(2)O(2). When wild-type H. polymorpha was grown on mixtures of glucose and methanol, the CCP level was independent of the rate of methanol utilization, whereas the level of catalase increased with increasing amounts of methanol in the substrate feed. Also, the wild type decomposed H(2)O(2) at a high rate when cells were grown on mixtures of glucose and H(2)O(2). In this case, an increase of both CCP and catalase was observed. When Saccharomyces cerevisiae was grown on mixtures of glucose and H(2)O(2), the level of catalase remained low, but CCP increased with increasing rates of H(2)O(2) utilization. From these observations and an analysis of cell yields under the various conditions, two conclusions can be drawn. (i) CCP is a key enzyme of H(2)O(2) detoxification in yeasts. (ii) Catalase can effectively compete with mitochondrial CCP for hydrogen peroxide only if hydrogen peroxide is generated at the site where catalase is located, namely in the peroxisomes. PMID- 16347720 TI - Effect of benzoic Acid on growth yield of yeasts differing in their resistance to preservatives. AB - Yeasts grown in the presence of benzoic acid tolerated 40 to 100% higher benzoic acid concentrations than did those grown in the absence of weak-acid-type preservatives. They also accumulated less benzoate in the presence of glucose. In chemostat cultures, benzoic acid reduced growth yield and the rate of cell production but increased specific fermentation rates. Benzoate contents were lower than those required for equilibrium when cells were impermeable to benzoate anion. Intracellular pHs were maintained near neutrality. Between species, stimulation of fermentation was inversely related to preservation resistance but was unrelated to the maximum rate of fermentation. The results show that a major effect of benzoic acid on yeasts in the presence of an energy source is the energy requirement for the reduction in cytoplasmic benzoate concentration and maintenance of pH. This energy source is unavailable for growth, resulting in lower growth yields and rates. Resistant species may be less permeable to undissociated benzoic acid. PMID- 16347721 TI - Sodium-Dependent Azotobacter chroococcum Strains Are Aeroadaptive, Microaerophilic, Nitrogen-Fixing Bacteria. AB - Na -dependent strains of Azotobacter chroococcum were observed to have very low reactivities with the H(2)O(2) spot test for catalase. The cell extract of the representative Na -dependent strain 184 contained a catalase specific activity that was 10-to 600-fold lower than those found in Na -independent strains of A. chroococcum. Peroxidase and superoxide dismutase activities existed in all strains, although only certain Na -dependent strains contained a peroxidase reactive with p-phenylenediamine. The activities of catalase and peroxidase in the Na -dependent strain 184 were dependent on iron availability, which helped to explain the iron-dependent growth characteristic of this strain. The activities of these enzymes were not increased by subjecting the cells to increased aeration, nitrogen-fixing conditions, or paraquat. Strain 184 was found to be very sensitive to H(2)O(2) or paraquat, even under iron-sufficient conditions, and was difficult to recover quantitatively on solid plating media. Strain 184 was more susceptible to H(2)O(2) when grown under low-aeration, nitrogen-fixing conditions than when it was grown in the presence of NH(4). Low population densities of strain 184 grew in nitrogen-free medium under microaerophilic conditions, while more dense populations were able to fix nitrogen under aerobic conditions. Therefore, these bacteria appeared to be aeroadaptive, microaerophilic, nitrogen-fixing bacteria. PMID- 16347722 TI - Mineralization of carbofuran by a soil bacterium. AB - A bacterium, tentatively identified as an Arthrobacter sp., was isolated from flooded soil that was incubated at 35 degrees C and repeatedly treated with carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl N-methylcarbamate). This bacterium exhibited an exceptional capacity to completely mineralize the ring labeled C in carbofuran to CO(2) within 72 to 120 h in a mineral salts medium as a sole source of carbon and nitrogen under aerobic conditions. Mineralization was more rapid at 35 degrees C than at 20 degrees C. No degradation of carbofuran occurred even after prolonged incubation under anaerobic conditions. The predicted metabolites of carbofuran, 7-phenol (2,3-dihydro-2,2-dimethyl-7 benzofuranol) and 3-hydroxycarbofuran, were also metabolized rapidly. 7-Phenol, although formed during carbofuran degradation, never accumulated in large amounts, evidently because of its further metabolism through ring cleavage. The bacterium readily hydrolyzed carbaryl (1-naphthyl N-methylcarbamate), but its hydrolysis product, 1-naphthol, resisted further degradation by this bacterium. PMID- 16347723 TI - Oxidation of Manganese and Formation of Mn(3)O(4) (Hausmannite) by Spore Coats of a Marine Bacillus sp. AB - Isolated spore coats of a marine Bacillus species were incubated in 25 mM MnCl(2) at pH 7.5. Manganese precipitates, formed on the coat surfaces, were analyzed by transmission electron microscopy, electron diffraction, and energy-dispersive X ray spectroscopy. Initially, an amorphous manganese oxide was observed on the coats which recrystallized to hausmannite after prolonged incubation in the MnCl(2) solution. The spore coats catalyze the oxidation of Mn(II) and have no structural influence on the final mineral phase precipitated. PMID- 16347724 TI - Improved method for determining bacterial filtration rates in zooplankton. AB - Filtration rates were determined for a natural population of zooplankton grazers (Bosmina longirostris [Mull.], Cyclops vicinus vicinus [Ulianine], Acanthodiaptomus denticornis [Wierz.], and Daphnia longispina [Mull.]) by using H labeled bacteria as food for these organisms. There was a relationship between filtration rates of the major zooplankton grazers and the prevailing algal and bacterial composition in the lake water. Low filtration rates were obtained in the presence of colonial and filamentous cyanobacteria. The rapid process of bacterial adhesion to the external organs of grazers can result in an overestimation of filtration rates. By using the simple method presented here, filtration rates, with simultaneous correction for bacterial adhesion, can be quickly determined. PMID- 16347725 TI - Induction and Catabolite Repression of beta-Glucosidase Synthesis in Myceliophthora thermophila D-14 (= ATCC 48104). AB - beta-Glucosidase activity in Myceliophthora thermophila D-14 (= ATCC 48104) was inducible and was produced in culture filtrate during growth with various inducers, of which PNPG (p-nitrophenyl-beta-d-glucoside) was the most efficient. Induction of beta-glucosidase also occurred when the organism was grown in medium supplemented with different carbon sources. Carboxymethyl cellulose, cellobiose, and Solka-Floc were found effective for induction of enzyme biosynthesis. The addition of glucose to the culture medium severely repressed beta-glucosidase synthesis, which could not be reversed by exogenous cyclic AMP or dibutyryl cyclic AMP. PMID- 16347726 TI - Microbial Degradation of n-Alkyl Tetrahydrothiophenes Found in Petroleum. AB - [This corrects the article on p. 1245 in vol. 54.]. PMID- 16347727 TI - Response in Soil of Cupriavidus necator and Other Copper-Resistant Bacterial Predators of Bacteria to Addition of Water, Soluble Nutrients, Various Bacterial Species, or Bacillus thuringiensis Spores and Crystals. AB - Soil was incubated with various species of bacteria, Bacillus subtilis, or Bacillus thuringiensis spores and crystals. These were added to serve as potential prey for indigenous, copper-resistant, nonobligate bacterial predators of bacteria in the soil. Alternatively, the soil was incubated with soluble nutrients or water only to cause potential indigenous prey cells to multiply so the predator cells would multiply. All of these incubation procedures caused excessive multiplication of some gram-negative bacteria in soil. Even greater multiplication, however, often occurred for certain copper-resistant bacterial predators of bacteria that made up a part of the gram-negative response. Incubation of the soil with copper per se did not give these responses. In most cases, the copper-resistant bacteria that responded were Cupriavidus necator, bacterial predator L-2, or previously unknown bacteria that resembled them. As was the case for C. necator and L-2, these new bacteria did not use glucose, had white colonies, produced copper-related growth initiation factor (GIF), and attacked B. thuringiensis spores on laboratory media. The results were different, however, when B. thuringiensis spores and crystals per se were added to the soil. The copper-resistant bacterial response in the soil did not, to any extent, include C. necator-like bacteria. Instead, the main copper-resistant bacterial predators that developed had yellow colonies and did not resemble C. necator or L 2 in other ways. They were not seen before, and they did not develop on the addition of B. subtilis spores to soil. Apparently, they could not produce a C. necator-like GIF. Nevertheless, they did respond very quickly to B. thuringiensis spores and crystals in soil, as if a GIF of some sort were involved. These results suggest that, under various conditions of soil incubation, gram-negative bacterial predators of bacteria multiply and that several copper-resistant types among them can be detected, counted, and isolated by plating dilutions of the soil onto media containing excess copper. PMID- 16347728 TI - Effect of pentoses and pentitols on fermentation of hay by mixed populations of ruminal microorganisms. AB - Consecutive batch culture, a technique which involves sequential transfer of cultures to fresh medium at regular intervals, was used to establish mixed ruminal-microbial populations in an anaerobic medium containing highly digestible hay. Once volatile fatty acid production was stable, perturbations were imposed in consecutive cultures by the addition of one of each of the following pentoses or analogous pentitols: l-arabinose, d-lyxose, d-ribose, d-xylose, l-arabitol, d arabitol (lyxitol), ribitol, and xylitol. With the exception of d-lyxose, the addition of pentoses caused marked increases in propionate and valerate production, and except for d-arabitol, pentitol addition caused increases in butyrate and valerate production. On transfer to and continued incubation in the control medium, volatile fatty acid production reverted to preperturbed levels. The presence of pentitols and pentoses significantly reduced the endpoint pH of cultures and the proportion of hay that was fermented. With all added substrates, the response to the perturbation was at its maximum within one incubation (i.e., within 48 h). Similarly, the variables being monitored all returned to control levels within one incubation. On the basis of these results, it is suggested that changes were related to the need to maintain a redox balance within anaerobic cultures rather than any significant changes in the microbial population that was present. PMID- 16347729 TI - Effects of Lactic Acid Bacteria and Organic Acids on Growth and Germination of Bacillus cereus. AB - Growth and germination of vegetative cells and endospores of Bacillus cereus were affected by Streptococcus lactis, Streptococcus thermophilus, Lactobacillus acidophilus, and Lactobacillus bulgaricus in nonfat milk medium and by salts of organic acids in broth medium. Growth of the lactic acid bacteria was not affected by B. cereus. B. cereus increased rapidly to about 10 CFU/ml when cells were added at the beginning of growth of lactic acid bacteria; it was inactivated slowly when added after 24 h and rapidly when added after 72 h of lactic acid bacterial growth. Streptococci were more inhibitory to the growth of B. cereus than lactobacilli were at 24 h. Spore germination was not affected after 24 h, but it was inhibited after 48 and 72 h of lactic acid bacterial growth. Acetate was more inhibitory to the growth of vegetative cells, while formate was more inhibitory to spore germination. Acetate, formate, and lactate (all at 0.1 M) completely inactivated multiplication of B. cereus at pH 6.1, 6.0, and 5.6, respectively. Spores of B. cereus were more resistant to these organic acids compared with the resistance of vegetative cells. Formate, lactate, and acetate (all at 0.1 M) caused 50% inhibition of spore germination at pH 4.4, 4.3, and 4.2, respectively. PMID- 16347730 TI - Plasmid frequency fluctuations in bacterial populations from chemically stressed soil communities. AB - The frequency of plasmids in chemically stressed bacterial populations was investigated by individually adding various concentration of kanamycin, ampicillin, and mercuric chloride to soil samples. Viable bacterial populations were enumerated, soil respiration was monitored for up to 6 weeks as an indicator of physiological stress, and bacterial isolates from stressed and control soils were screened for the presence of plasmids. Low levels of the chemical stress factors did not for the most part significantly alter population viability, soil respiration, or plasmid frequency. Exposure to high stress levels of mercury and ampicillin, however, resulted in altered numbers of viable organisms, soil respiration, and plasmid frequency. Plasmid frequency increased in response to ampicillin exposure but was not significantly changed after exposure to kanamycin. In mercuric chloride-stressed soils, there was a decrease in plasmid frequency despite an increase in overall mercury resistance of the isolates, suggesting that mercury resistance in these populations is largely, if not completely, chromosome encoded. Chemical stress did not cause an increase in plasmid-mediated multiple resistance. A genetic response (change in plasmid frequency) was not found unless a physiological (phenotypic) response (change in viable cells and respiratory activity) was also observed. The results indicate that a change in plasmid frequency is dependent on both the amount and type of chemical stress. PMID- 16347731 TI - Biodegradation of nitriles in shale oil. AB - Enrichment cultures were obtained, after prolonged incubation on a shale oil as the sole source of nitrogen, that selectively degraded nitriles. Capillary gas chromatographic analyses showed that the mixed microbial populations in the enrichments degraded the homologous series of aliphatic nitriles but not the aliphatic hydrocarbons, aromatic hydrocarbons, or heterocyclic-nitrogen compounds found in this oil. Time course studies showed that lighter nitriles were removed more rapidly than higher-molecular-weight nitriles. A Pseudomonas fluorescens strain isolated from an enrichment, which was able to completely utilize the individual nitriles undecyl cyanide and undecanenitrile as sole sources of carbon and nitrogen, was unable to attack stearonitrile when provided alone as the growth substrate. A P. aeruginosa strain, also isolated from one of the enrichments, used nitriles but not aliphatic or aromatic hydrocarbons when the oil was used as a sole nitrogen source. However, when the shale oil was used as the sole source of carbon, aliphatic hydrocarbons in addition to nitriles were degraded but aromatic hydrocarbons were still not attacked by this P. aeruginosa strain. PMID- 16347732 TI - Demethylation of dimethylsulfoniopropionate and production of thiols in anoxic marine sediments. AB - Dimethylsulfoniopropionate (DMSP) is a natural product of algae and aquatic plants, particularly those from saline environments. We investigated whether DMSP could serve as a precursor of thiols in anoxic coastal marine sediments. The addition of 10 or 60 muM DMSP to anoxic sediment slurries caused the concentrations of 3-mercaptopropionate (3-MPA) and methanethiol (MSH) to increase. Antibiotics prevented the appearance of these thiols, indicating biological formation. Dimethyl sulfide (DMS) and acrylate also accumulated after the addition of DMSP, but these compounds were rapidly metabolized by microbes and did not reach high levels. Acrylate and DMS were probably generated by the enzymatic cleavage of DMSP. MSH arose from the microbial metabolism of DMS, since the direct addition of DMS greatly increased MSH production. Additions of 3 methiolpropionate gave rise to 3-MPA at rates similar to those with DMSP, suggesting that sequential demethylation of DMSP leads to 3-MPA formation. Only small amounts of MSH were liberated from 3-methiolpropionate, indicating that demethiolation was not a major transformation for 3-methiolpropionate. We conclude that DMSP was degraded in anoxic sediments by two different pathways. One involved the well-known enzymatic cleavage to acrylate and DMS, with DMS subsequently serving as a precursor of MSH. In the other pathway, successive demethylations of the sulfur atom proceeded via 3-methiolpropionate to 3-MPA. PMID- 16347733 TI - Calculation of cell production from [h]thymidine incorporation with freshwater bacteria. AB - The conversion factor for the calculation of bacterial production from rates of [H]thymidine incorporation was examined with diluted batch cultures of freshwater bacteria. Natural bacterial assemblages were grown in aged, normal, and enriched media at 10 to 20 degrees C. The generation time during 101 growth cycles covered a range from 4 to >200 h. The average conversion factor was 2.15 x 10 cells mol of thymidine incorporated into the trichloroacetic acid (TCA) precipitate (standard error = 0.29 x 10; n = 54), when the generation time exceeded 20 h. At generation times of <20 h, the average conversion factor was 11.8 x 10 cells mol of thymidine incorporated into TCA precipitate (standard error = 1.72 x 10; n = 47). The amount of radioactivity in purified DNA increased with decreasing generation time and increasing conversion factor (calculated from the TCA precipitate), corresponding to a decrease in the percentage in protein. The conversion factors calculated from purified DNA or from the TCA precipitate gave the same variability. Conversion factors did not change significantly with the medium, but were significantly higher at 20 degrees C than at 15 and 10 degrees C. A detailed examination of the [H]thymidine concentrations that were needed to achieve maximum labeling in DNA was carried out 6 times during a complete growth cycle. During periods with low generation times and high conversion factors, 15 nM [H]thymidine was enough for the maximum labeling of the TCA precipitate. This suggests that incorporation of [H]thymidine into DNA is probably limited by uptake during periods with generation times of <20 h and that freshwater bacterioplankton cell production sometimes is underestimated when a conversion factor of 2.15 x 10 cells mol of thymidine incorporated is used. PMID- 16347734 TI - Heat Production by the Denitrifying Bacterium Pseudomonas fluorescens and the Dissimilatory Ammonium-Producing Bacterium Pseudomonas putrefaciens during Anaerobic Growth with Nitrate as the Electron Acceptor. AB - The heat production rate and the simultaneous nitrate consumption and production and consumption of nitrite and nitrous oxide were monitored during the anaerobic growth of two types of dissimilatory nitrate reducers. Pseudomonas fluorescens, a denitrifier, consumed nitrate and accumulated small amounts of nitrite or nitrous oxide. The heat production rate increased steadily during the course of nitrate consumption and decreased rapidly concomitant with the depletion of the electron acceptors. A mean experimental enthalpy change value of -800 kJ/mol of nitrate and a mean growth yield value of 33 g (dry weight)/mol of nitrate consumed were obtained for different concentrations of nitrate. For Pseudomonas putrefaciens, a dissimilatory ammonium producer, the nitrate consumption resulted in an accumulation of nitrite and nitrous oxide. Nitrite consumption commenced after depletion of the nitrate; consequently, two phases were noted in the heat production rate curve during growth. A mean experimental enthalpy change value of -810 kJ/mol of nitrate was obtained for different concentrations of nitrate. PMID- 16347735 TI - Transformations of halogenated aromatic aldehydes by metabolically stable anaerobic enrichment cultures. AB - Metabolically stable enrichment cultures of anaerobic bacteria obtained by elective enrichment of sediment samples from the Baltic Sea and Gulf of Bothnia have been used to study the oxidation and reduction of the aldehyde group of various halogenated aromatic aldehydes. During the transformation of 5- and 6 chlorovanillin, 6-bromovanillin, 3-chloro-4-hydroxybenzaldehyde, 3,5-dichloro-4 hydroxybenzaldehyde, and 3,5-dibromo-4-hydroxybenzaldehyde, it was shown that synthesis of the corresponding carboxylic acids, which were the principal metabolites, was invariably accompanied by partial reduction of the aldehyde to a hydroxymethyl group in yields of between 3 and 30%. Complete reduction to a methyl group was observed with some of the halogenated vanillins, but to an extremely limited extent with the halogenated 4-hydroxybenzaldehydes. One consortium produced both the hydroxymethyl and methyl compounds from both 5- and 6-chlorovanillin: it was therefore assumed that the methyl compound was the ultimate reduction product. On the basis of the kinetics of formation of the metabolites, it was concluded that the oxidation and reduction reactions were mechanistically related. In addition to these oxidations and reductions, dehalogenation was observed with one of the consortia. In contrast to the transformations of 5- and 6-chlorovanillin, which produced chlorinated methylcatechols, the corresponding compounds were not observed with 5- and 6 bromovanillin: the former was debrominated, forming 4-methylcatechol, whereas the latter produced 6-bromovanillyl alcohol without demethylation. Similarly, although 3-chloro-4-hydroxybenzaldehyde formed the chlorinated carboxylic acid and the benzyl alcohol, the 3-bromo compound was debrominated with formation of 4 hydroxybenzoic acid and, ultimately, phenol. On prolonged incubation, the halogenated carboxylic acids were generally decarboxylated, so that the final products from these substrates were halogenated catechols or phenols. Reductive processes of the type revealed in this study might therefore plausibly occur in the environment during anaerobic transformation of halogenated aromatic aldehydes containing hydroxyl and/or methoxyl groups. PMID- 16347736 TI - Production by Streptomyces viridosporus T7A of an Enzyme Which Cleaves Aromatic Acids from Lignocellulose. AB - The lignocellulose-degrading actinomycete Streptomyces viridosporus T7A produced an extracellular esterase when grown in a mineral salts-yeast extract medium. Extracellular esterase activity was first detected during the late stationary phase and typically followed the appearance of intracellular activity. When the organism was grown in lignocellulose-supplemented medium, esterase activity was not increased, but lignocellulose-esterified p-coumaric acid and vanillic acid were released into the medium. Polyacrylamide gels showed that several extracellular esterases differing in substrate specificity were produced. Ultrafiltration was used to concentrate the esterase prior to purification. Activity was recovered mostly in the molecular weight fraction between 10,000 and 100,000. Concentrated esterase was further purified by DEAE-Sepharose anion exchange chromatography to a specific activity 11.82 times greater than that in the original supernatant. There were seven detectable esterase active proteins in the partially purified enzyme solution. Three were similar esterases that may be isoenzymes. The partially purified esterase had a pH optimum for activity of 9.0, a temperature optimum of 45 to 50 degrees C, and a K(m) and V(max) of 0.030 mM and 0.097 mumol/min per ml, respectively, when p-nitrophenyl butyrate was the substrate. The enzyme was unstable above 40 degrees C but retained activity when stored at 4 or -20 degrees C. It lost some activity (20%) when lyophilized. Substrate specificity assays showed that it hydrolyzed ester linkages of p nitrophenyl butyrate, alpha-naphthyl acetate, alpha-naphthyl butyrate, and lignocellulose. Vanillic and p-coumaric acids were identified as products released from lignocellulose. The enzyme is thought to be a component of the lignocellulose-degrading enzyme system of S. viridosporus. PMID- 16347737 TI - Combined oxygen and nitrous oxide microsensor for denitrification studies. AB - The construction of a microsensor which can be used to measure O(2) and N(2)O simultaneously is described. The microsensor exhibited a linear response to both O(2) and N(2)O, and the response to N(2)O was independent of the O(2) concentration and vice versa. The N(2)O detection limit of a microsensor with a tip diameter of 20 mum was around 1 mumol liter. The signals for O(2) and N(2)O were affected by hydrogen sulfide, but other interfering agents were not observed in the biofilms and sediments analyzed. Microprofiles of O(2) and N(2)O were measured in a biofilm which was exposed to acetylene to block the N(2)O reductase activity of denitrifying bacteria. O(2) penetrated about 0.5 mm into the biofilm and was not affected by acetylene, but the N(2)O concentration at 1.4 mm depth increased from 32 to 411 mumol liter after the addition of the inhibitor. The shape of the N(2)O profile after the addition of acetylene showed that denitrification (denitrifying activity) was detectable in all anoxic layers of the biofilm. PMID- 16347738 TI - Monoclonal Antibodies to the Cell-Wall-Associated Proteinase of Lactococcus lactis subsp. cremoris Wg2. AB - Twelve monoclonal antibodies directed to the cell-wall-associated proteinase of Lactococcus lactis subsp. cremoris Wg2 were isolated after immunization of BALB/c mice with a partially purified preparation of the proteinase. The monoclonal antibodies reacted with the 126-kilodalton proteinase band in a Western immunoblot. All but one of the monoclonal antibodies reacted with protein bands with a molecular weight below 126,000, possibly degradation products of the proteinase. The monoclonal antibodies could be divided into six groups according to their different reactions with the proteinase degradation products in the Western blot. Different groups of monoclonal antibodies reacted with different components of the L. lactis subsp. cremoris Wg2 proteinase. Crossed immunoelectrophoresis showed that monoclonal antibody groups I, II, and III react with proteinase component A and that groups IV, V, and VI react with proteinase component B. The isolated monoclonal antibodies cross-reacted with the proteinases of other L. lactis subspecies. Monoclonal antibodies of group IV cross-reacted with proteinase component C of other L. lactis subsp. cremoris strains. The molecular weight of the proteinase attached to the cells of L. lactis subsp. cremoris Wg2 was 200,000, which is different from the previously reported values. This could be analyzed by immunodetection of the proteinase on a Western blot. This value corresponds to the molecular weight calculated from the amino acid sequence of the cloned L. lactis subsp. cremoris Wg2 proteinase gene. PMID- 16347739 TI - Microbiologically catalyzed enantio- and diastereoselective oxidation of chrysanthemol stereoisomers to chrysanthemic acids. AB - The diastereo- and enantioselective microbial oxidation of a mixture of racemic cis/trans-chrysanthemols to the corresponding stereoisomeric chrysanthemic acids by Aspergillus species is described. Of the three microorganisms which were found capable of oxidizing racemic cis/trans-chrysanthemols, A. ochraceus ATCC 18500 showed complete enantioselectivity for (+)-stereoisomers [(+)-trans-chrysanthemol and (+)-cis-chrysanthemol), whereas A. flavipes ATCC 1030 and ATCC 11013 showed complete enantioselectivity for the (+)-cis-chrysanthemol but a time-dependent enantioselectivity during oxidation of trans-chrysanthemol [oxidation of (+) trans-chrysanthemol prior to (-)-trans-chrysanthemol]. The diastereoselectivity of all three microorganisms was time dependent, in that the trans-stereoisomers were oxidized prior to the cis-isomers. PMID- 16347740 TI - Host Ranges of the IncN Group Plasmid pCU1 and Its Minireplicon in Gram-Negative Purple Bacteria. AB - The bacterial host ranges of the conjugatively self-transmissible IncN group plasmid pCU1 and its mobilizable miniderivative, pCU785, were examined. Species of the gram-negative purple bacteria were chosen for this study. Conjugative mobilization of pCU785 into a wide variety of bacteria was facilitated by the presence of oriT of the broad-host-range plasmid RK2 in pCU785. Although the host range of the IncN tra system of pCU1 is broad, the host range of its replicon is limited. However, the pCU1 replicon can be maintained in Agrobacterium, Bradyrhizobium, and Rhizobium species under conditions that select for plasmid maintenance. It is lost efficiently from these populations on release of selection. PMID- 16347741 TI - Effect of Plant Species and Environmental Conditions on Ice Nucleation Activity of Pseudomonas syringae on Leaves. AB - Selected plant species and environmental conditions were investigated for their influences on expression of ice nucleation activity by 15 Pseudomonas syringae strains grown on plants in constant-temperature growth chamber studies. Ice nucleation frequencies (INFs), the fraction of cells that expressed ice nucleation at -5 or -9 degrees C, of individual strains varied greatly, both on plants and in culture. This suggests that the probability of frost injury, which is proportional to the number of ice nuclei on leaf surfaces, is strongly determined by the particular bacterial strains that are present on a leaf surface. The INFs of strains were generally higher when they were grown on plants than when they were grown in culture. In addition, INFs in culture did not correlate closely with INFs on plants, suggesting that frost injury prediction should be based on INF measurements of cells grown on plants rather than in culture. The relative INFs of individual strains varied with plant host and environment. However, none of seven plant species tested optimized the INFs of all 15 strains. Similarly, incubation for 48 h at near 100% relative humidity with short photoperiods did not always decrease the INF when compared with a 72 h, 40% relative humidity, long-photoperiod incubation. Pathogenic strains on susceptible hosts were not associated with higher or lower INFs relative to their INFs on nonsusceptible plant species. The ice nucleation activity of individual bacterial strains on plants therefore appears to be controlled by complex and interacting factors such as strain genotype, environment, and host plant species. PMID- 16347742 TI - Production of alpha-Amylase by the Ruminal Anaerobic Fungus Neocallimastix frontalis. AB - alpha-Amylase production was examined in the ruminal anaerobic fungus Neocallimastix frontalis. The enzyme was released mainly into the culture fluid and had temperature and pH optima of 55 degrees C and 5.5, respectively, and the apparent K(m) for starch was 0.8 mg ml. The products of alpha-amylase action were mainly maltotriose, maltotetraose, and longer-chain oligosaccharides. No activity of the enzyme was observed towards these compounds or pullulan, but activity on amylose was similar to starch. Evidence for the endo action of alpha-amylase was also obtained from experiments which showed that the reduction in iodine-staining capacity and release in reducing power by action on amylose was similar to that for commercial alpha-amylase. Activities of alpha-amylase up to 4.4 U ml (1 U represents 1 mumol of glucose equivalents released per min) were obtained for cultures grown on 2.5 mg of starch ml in shaken cultures. No growth occurred in unshaken cultures. With elevated concentrations of starch (>2.5 mg ml), alpha amylase production declined and glucose accumulated in the cultures. Addition of glucose to cultures grown on low levels of starch, in which little glucose accumulated, suppressed alpha-amylase production, and in bisubstrate growth studies, active production of the enzyme only occurred during growth on starch after glucose had been preferentially utilized. When cellulose, cellobiose, glucose, xylan, and xylose were tested as growth substrates for the production of alpha-amylase (initial concentration, 2.5 mg ml), they were found to be less effective than starch, but maltose was almost as effective. The fungal alpha amylase was found to be stable at 60 degrees C in the presence of low concentrations of starch ( 300 nm). The results reinforce recent evidence for the broad potential of aromatic hydroxylase systems for the destruction of pollutants. PMID- 16347747 TI - Microbial Conversion of alpha-Ionone, alpha-Methylionone, and alpha Isomethylionone. AB - alpha-Ionone, alpha-methylionone, and alpha-isomethylionone were converted by Aspergillus niger JTS 191. The individual bioconversion products from alpha ionone were isolated and identified by spectrometry and organic synthesis. The major products were cis-3-hydroxy-alpha-ionone, trans-3-hydroxy-alpha-ionone, and 3-oxo-alpha-ionone. 2,3-Dehydro-alpha-ionone, 3,4-dehydro-beta-ionone, and 1-(6,6 dimethyl-2-methylene-3-cyclohexenyl)-buten-3-one were also identified. Analogous bioconversion products from alpha-methylionone and alpha-isomethylionone were also identified. From results of gas-liquid chromatographic analysis during the fermentation, we propose a metabolic pathway for alpha-ionones and elucidation of stereochemical features of the bioconversion. PMID- 16347748 TI - Pressure-Induced Alterations in the Protein Pattern of the Thermophilic Archaebacterium Methanococcus thermolithotrophicus. AB - Elevated hydrostatic pressure has been shown to affect the growth rate of the thermophilic methanobacterium Methanococcus thermolithotrophicus without extending its temperature range of viability. Analysis of the cell inventory after approximately 10 h of incubation at 65 degrees C and 50 MPa (applying high pressure liquid chromatography and two-dimensional gel electrophoresis) proved that pressure induces alterations in the protein pattern and the amino acid composition of the total cell hydrolysate. Gels showed that after pressurization a series of (basic) proteins with a molecular mass in the range of 38 and 70 kilodaltons occurs which is not detectable in cells grown at normal atmospheric pressure. The question of whether the observed alterations are caused by the perturbation of the balance of protein synthesis and turnover or by the pressure induced synthesis of compounds analogous to heat shock proteins remains unanswered. PMID- 16347749 TI - Production of an Extracellular Polysaccharide by Haloferax mediterranei. AB - The extremely halophilic archaebacterium Haloferax mediterranei produces an exocellular polymeric substance that gives the colonies a typical mucous character and is responsible for the appearance of a superficial layer in unshaken liquid medium. This exocellular polymeric substance can be obtained from the supernatant of shaken liquid cultures by cold ethanol precipitation, and yields as high as 3 mg/ml have been detected. The substance was produced under all the conditions tested and with all substrates assayed, although higher yields were obtained with sugars, particularly glucose, as carbon and energy source. The total exocellular polymeric substance produced was proportional to the total biomass. The polymer is a heteropolysaccharide containing mannose as the major component. Glucose, galactose, and another unidentified sugar were also present, as well as amino sugars, uronic acids, and a considerable amount of sulfate, which accounts for the acidic nature of the polymer. The infrared spectrum and specific assays showed the absence of acyl groups. The rheological properties of polymer solutions were studied, showing a pseudoplastic behavior and a high apparent viscosity at relatively low concentrations. Viscosity was remarkably resistant to extremes of pH, temperature, or salinity. These characteristics make this polymer interesting for enhanced oil recovery and other applications for which a very resistant thickening agent is required. PMID- 16347750 TI - Effects of Bradyrhizobium japonicum and Soybean (Glycine max (L.) Merr.) Phosphorus Nutrition on Nodulation and Dinitrogen Fixation. AB - Cells of Bradyrhizobium japonicum were grown in media containing either 1.0 mM or 0.5 muM phosphorus. In growth pouch experiments, infection of the primary root of soybean (Glycine max (L.) Merr.) by B. japonicum USDA 31, 110, and 142 was significantly delayed when P-limited cells were applied to the root. In a greenhouse experiment, B. japonicum USDA 31, 110, 122, and 142 grown with sufficient and limiting P were used to inoculate soybeans which were grown with either 5 muM or 1 mM P nutrient solution. P-limited cells of USDA 31 and 110 formed significantly fewer nodules than did P-sufficient cells, but P-limited cells of USDA 122 and 142 formed more nodules than P-sufficient cells. The increase in nodule number by P-limited cells of USDA 142 resulted in significant increases in both nodule mass and shoot total N. In plants grown with 1 mM P, inoculation with P-limited cells of USDA 110 resulted in lower total and specific nitrogenase activities than did inoculation with P-sufficient cells. Nodule numbers, shoot dry weights, and total N and P were all higher in plants grown with 1 mM P, and plants inoculated with USDA 31 grew poorly relative to plants receiving strains USDA 110, 122, and 142. Although the effects of soybean P nutrition were more obvious than those of B. japonicum P nutrition, we feel that it is important to develop an awareness of the behavior of the bacterial symbiont under conditions of nutrient limitation similar to those found in many soils. PMID- 16347751 TI - Product inhibition of butyrate metabolism by acetate and hydrogen in a thermophilic coculture. AB - Studies on product inhibition of a thermophilic butyrate-degrading bacterium in syntrophic association with Methanobacterium thermoautotrophicum showed that a gas phase containing more than 2 x 10 atm (2.03 kPa) of hydrogen prevented growth and butyrate consumption, while a lower hydrogen partial pressure of 1 x 10 to 2 x 10 atm (0.1 to 2.03 kPa) gradually inhibited the butyrate consumption of the coculture. No inhibition of butyrate consumption was found on the addition of 0.75 x 10 atm (76 Pa) of hydrogen to the gas phase. A slight inhibition of butyrate consumption by the coculture occurred at an acetate concentration of 16.4 mM. Inhibition gradually increased with increasing acetate concentration up to 81.4 mM, when complete inhibition of butyrate consumption occurred. When the culture contained an acetate-utilizing methanogen in addition to M. thermoautotrophicum, the inhibition of the triculture by acetate was gradually reversed as the acetate concentration was lowered by the aceticlastic methanogen. The results show that optimal growth conditions for the thermophilic butyrate degrading bacterium depend on both hydrogen and acetate removal. PMID- 16347752 TI - Effect of Nitrogen on Polysaccharide Production in a Porphyridium sp. AB - Porphyridium cultures grown on either nitrate or ammonium as the nitrogen source showed similar patterns of growth and cell wall polysaccharide production. The effect of nitrogen on growth and cell wall polysaccharide production was studied by applying three regimens of supply: batch mode, in which nitrate was supplied at the beginning of the experiment and became depleted at day 6; continual mode, in which nitrate was added daily; and deficient mode, in which the cells were cultured in a nitrate-free medium. Growth was similar in the batch- and continual mode cultures, whereas it was totally inhibited in the deficient-mode culture. Polysaccharide content (per volume) was highest in the batch-mode culture and lowest in the deficient-mode culture. However, polysaccharide production per cell was similar in the continual- and deficient-mode cultures, the highest value being found in the batch-mode culture. In addition to its effect on polysaccharide content, nitrogen affected the polysaccharide distribution between soluble and bound polysaccharides. In the deficientmode culture, most of the cell wall polysaccharide was dissolved in the medium. PMID- 16347753 TI - Degradation of 2-methylisoborneol by aquatic bacteria. AB - 2-Methylisoborneol (MIB) is a musty- or muddy-smelling compound which occurs in some natural waters and which is difficult to remove by conventional water treatment methods. Bacterial degradation of MIB was examined in batch culture experiments. Cultures able to metabolize MIB were enriched in a mineral salts medium supplemented with milligram-per-liter levels of the compound and were inoculated with water and sediment samples from reservoirs where MIB is seasonally produced. Bacteria from degrading cultures were isolated on R2A agar and identified as predominantly Pseudomonas spp. Degradation occurred only in cultures consisting of three or more different bacteria. MIB supported growth as the sole added carbon source at 1 to 6.7 mg/liter. MIB was also degraded at microgram-per-liter levels in sterile filtered lake water inoculated with washed bacteria and in synthetic medium supplemented with various sugars or acetate. Complete degradation of MIB took from 5 days to more than 2 weeks. Enrichment with isoborneol, a structural analog of MIB, failed as a preenrichment for MIB degraders. Isoborneol at 20 to 40 mg/liter readily supported bacterial growth, whereas MIB at 12 to 20 mg/liter took months to degrade. The relative recalcitrance of MIB compared with isoborneol may be a result of the additional methyl group in MIB. PMID- 16347754 TI - Influence of Calcium Ion on Ethanol Tolerance of Saccharomyces bayanus and Alcoholic Fermentation by Yeasts. AB - The addition of Ca (as CaCl(2)) in optimal concentrations (0.75 to 2.0 mM) to a fermentation medium with a trace contaminating concentration of Ca (0.025 mM) led to the rapid production of higher concentrations of ethanol by Saccharomyces cerevisiae, Saccharomyces bayanus, and Kluyveromyces marxianus. The positive effect of calcium supplementation (0.75 mM) on alcoholic fermentation by S. bayanus was explained by the increase in its ethanol tolerance. The ethanol inhibition of growth and fermentation followed the equation mu(xi) = mu(oi) [1 - (X/X(mi))], where mu(oi) and mu(xi) are, respectively, the specific growth (i = g) and fermentation (i = f) rates in the absence or presence of a concentration (X) of added ethanol, and X(mi) is the maximal concentration of ethanol which allows growth or fermentation. The toxic power is given by n(i). In Ca - supplemented medium (0.75 mM), n(g) = 0.42 for growth and n(f) = 0.43 for fermentation compared with 0.52 and 0.55, respectively, in unsupplemented medium; for both media, X(mg) = 10% (vol/vol) and X(mf) = 13% (vol/vol). For lethal concentrations of ethanol, the specific death rates were minimal for cells that were grown and incubated with ethanol in medium with an optimal concentration of Ca, maximal for cells grown and incubated with ethanol in unsupplemented medium, and intermediate for cells grown in unsupplemented medium and incubated with ethanol in calcium-supplemented medium. The effect of Ca on the acidification curve of energized cells in the presence of ethanol was found to be closely associated with its protective effect on growth, fermentation, and viability. PMID- 16347755 TI - New amylolytic yeast strains for starch and dextrin fermentation. AB - Yeast strains capable of fermenting starch and dextrin to ethanol were isolated from samples collected from Brazilian factories in which cassava flour is produced. Considerable alcohol production was observed for all the strains selected. One strain (DI-10) fermented starch rapidly and secreted 5 times as much amylolytic enzyme than that observed for Schwanniomyces alluvius UCD 54-83. This strain and three other similar isolates were classified as Saccharomyces cerevisiae var. diastaticus by morphological and physiological characteristics and molecular taxonomy. PMID- 16347756 TI - Fruiting at High Temperature and Its Genetic Control in the Basidiomycete Flammulina velutipes. AB - Of 10 geographic strains of Flammulina velutipes, 4 were found capable of fruiting at 22 degrees C (Fr(H)) rather than at the typical 15 degrees C (Fr(L)). Crosses made between Fr(H) and Fr(L) monokaryons were never observed to fruit at 22 degrees C. However, some hybrids did fruit at the intermediate temperature of 18 degrees C when grown on appropriate substrates, indicating incomplete dominance of the low-temperature requirement. Analysis of progeny of five Fr(H) x Fr(L) crosses indicated that a minimum of two genes appears to control the requirement for fruiting at 1.07 mumol mg of fungal dry weight. At levels below this, protease synthesis was initiated; subsequently, free amino nitrogen appeared in the medium and diauxic growth was observed. Slow feeding with GlcNAc (35.34 mug ml h) did not repress protease synthesis nor did GlcNAc accumulate in the medium above 0.5 mg ml. Increasing the rate of release of GlcNAc (83.51 mug ml h) resulted in the accumulation of GlcNAc in the medium to 2.0 mg ml, a 45% increase in growth and a decrease in protease synthesis by about 81%. Free amino acids generated from the hydrolysis of gelatin did not repress protease synthesis. These data are interpreted in terms of known interaction of B. bassiana with insect cuticular components. We suggest that the action of extracellular chitinases synthesized by B. bassiana on insect cuticle, and pursuant release of GlcNAc, may have important consequences on the regulation of other extracellular catabolic enzymes such as the protease. PMID- 16347773 TI - Dynamics of Soil Denitrifier Populations: Relationships between Enzyme Activity, Most-Probable-Number Counts, and Actual N Gas Loss. AB - To better understand temporal variability in soil denitrification, denitrifying enzyme activity (DEA) and denitrifier populations (as determined by most-probable number [MPN] counts) were measured in field and laboratory experiments. Measurements of DEA and MPN provided highly contradictory indications of denitrifier dynamics. In laboratory incubations, under conditions favoring active denitrification, the synthesis of new denitrifying enzymes and the actual amount of denitrification were closely related. In other experiments, however, both DEA and MPN counts were poor indicators of actual denitrification. In some cases, we found significant increases in DEA but no significant production of N gas. Except with unnaturally high substrate amendments, changes in DEA were small relative both to the persistently high DEA background and to changes in MPN. As estimated by MPN counts, denitrifier populations increased significantly during denitrification events. It was apparent that only a small fraction of the denitrifiers were included in the MPN counts, but it appeared that this isolatable fraction increased during periods of active denitrifier growth. Use of DEA as an index of biomass of cells which have synthesized denitrifying enzymes suggested that denitrifier populations were persistent, stable, and much larger than indicated by MPN procedures. PMID- 16347774 TI - Thiolase from Clostridium acetobutylicum ATCC 824 and Its Role in the Synthesis of Acids and Solvents. AB - Thiolase (acetyl-coenzyme A [CoA] acetyltransferase, E.C. 2.3.1.19) from Clostridium acetobutylicum ATCC 824 has been purified 70-fold to homogeneity. Unlike the thiolase in Clostridium pasteurianum, this thiolase has high relative activity throughout the physiological range of internal pH of 5.5 to 7.0, indicating that change in internal pH during acid production is not an important factor in the regulation of this thiolase. In the condensation direction, the thiolase is inhibited by micromolar levels of CoA, and this may be an important factor in modulating the net condensation of acetyl-CoA to acetoacetyl-CoA. Other cofactors and metabolites that were tested and shown to be inhibitors are ATP and butyryl-CoA. The native enzyme consists of four 44,000-molecular-weight subunits. The kinetic binding mechanism is ping-pong. The K(m) value for acetyl-CoA is 0.27 mM at 30 degrees C and pH 7.4. The K(m) values for sulfhydryl-CoA and acetoacetyl CoA are, respectively, 0.0048 and 0.032 mM at 30 degrees C and pH 8.0. The active site apparently contains a sulfhydryl group, but unlike other thiolases, this thiolase is relatively stable in the presence of 5,5'-dithiobis(2-nitrobenzoic acid). Studies of thiolase specific activity under various types of continuous fermentations show that regulation of this enzyme at both the genetic and enzyme levels is important. PMID- 16347775 TI - Influence of ethylene produced by soil microorganisms on etiolated pea seedlings. AB - There is indirect evidence that soil microorganisms producing ethylene (C(2)H(4)) can influence plant growth and development, but unequivocal proof is lacking in the literature. A laboratory study was conducted to demonstrate the validity of this speculation. Four experiments were carried out to observe the characteristic "triple" response of etiolated pea seedlings to C(2)H(4) microbially derived from l-methionine as a substrate in the presence or absence of Ag(I), a potent inhibitor of C(2)H(4) action. In two experiments, the combination of l-methionine and Acremonium falciforme (as an inoculum) was used, while in another study the indigenous soil microflora was responsible for C(2)H(4) production. A standardized experiment was conducted with C(2)H(4) gas to compare the contribution of the microflora to plant growth. In all cases, etiolated pea seedlings exhibited the classical triple response, which includes reduction in elongation, swelling of the hypocotyl, and a change in the direction of growth (horizontal). The presence of Ag(I) afforded protection to the pea seedlings against the microbially derived C(2)H(4). This study demonstrates that microbially produced C(2)H(4) in soil can influence plant growth. PMID- 16347776 TI - Production of d-Aminoacylase from Alcaligenes denitrificans subsp. xylosoxydans MI-4. AB - A bacterial strain that produces d-aminoacylase was isolated from soil and identified as Alcaligenes denitrificans subsp. xylosoxydans MI-4. l-Aminoacylase activity in this strain was only 1 to 2% of d-aminoacylase activity. d Aminoacylase was inducibly produced. N-Acetyl-dl-leucine was the best inducer, and the d-isomer had the ability to induce the enzyme. Enzymatic resolution of N acetyl-dl-methionine with the crude enzyme was carried out, and the d/l ratio in the resolved methionine was approximately 100/7, suggesting that resolution with crude enzymes may become possible by removing small amounts of the contaminated l form with l-amino acid oxidase. PMID- 16347777 TI - Characterization of an Extracellular Protease from the Insect Pathogen Xenorhabdus luminescens. AB - Xenorhabdus luminescens Hm cultured in gelatin broth produced a single extracellular protease. The protease was purified by a factor of 500 and characterized as a monomeric protein with an approximate molecular weight of 61,000. On the basis of inhibitor studies and its pH optimum, the protease was classified as an alkaline metalloprotease with a pH optimum near 8; the isoelectric point of the enzyme is 4.2 +/- 0.2. The protease may be a major factor in the ecology of X. luminescens, which is carried as a symbiom of some parasitic nematodes. PMID- 16347778 TI - Identification of Erwinia amylovora, the Fireblight Pathogen, by Colony Hybridization with DNA from Plasmid pEA29. AB - All strains of Erwinia amylovora characterized carry a medium-size plasmid of 29 kilobases (pEA29). We mapped this plasmid with various restriction enzymes, cloned the whole DNA into an Escherichia coli plasmid, and subcloned restriction fragments. These DNA species were used for identification of E. amylovora after handling of strains in the laboratory and also in field isolates. About 70 strains of E. amylovora and 24 strains from nine other species, mainly found in plant habitats, were checked in a colony hybridization test. Virulent and avirulent E. amylovora strains reacted positively, whereas the other species were negative. Apart from the hybridization assay, the positive strains were additionally tested for ooze production on rich agar with 5% sucrose and on immature-pear slices. Unspecific background hybridization of non-E. amylovora strains found for hybridization with the whole E. amylovora plasmid was almost eliminated when a 5-kilobase SalI fragment from pEA29 was used as a probe and when the washes after the hybridization procedure were done with high stringency. Under these conditions, E. amylovora could be readily identified from field isolates. PMID- 16347779 TI - Properties of the Cell Walls of Lactococcus lactis subsp. cremoris SK110 and SK112 and Their Relation to Bacteriophage Resistance. AB - Resistance of Lactococcus lactis subsp. cremoris SK110 to bacteriophage sk11G, encoded on the plasmid pSK112, is due to poor phage adsorption. Its phage sensitive variant SK112, cured of pSK112, adsorbs phages effectively. Incubation of SK112 with concanavalin A remarkably reduced phage adsorption to this strain. This treatment also caused agglutination of SK112 that was not found with SK110, indicating different concanavalin A adsorption characteristics of cell walls of both strains. The differences between the two strains were reduced by a mild alkali treatment of cells. This resulted in a positive agglutination with concanavalin A for both strains and in parallel adsorption of phage sk11G to both. Moreover, isolated cell walls of the two strains were investigated, and both bound phage sk11G. These observations suggest the presence of phage receptor material in SK112 as well as in SK110. SK110 contained a relatively high level of bound galactose when compared with the phage-sensitive SK112. After the mild alkali treatment, however, the galactose content of SK110 was diminished such that it became comparable with that of SK112. It is hypothesized that the alkali treatment liberates a galactose-containing component from the cell wall and causes phage sensitivity in L. lactis subsp. cremoris SK110. PMID- 16347780 TI - Simultaneous Nitrification and Denitrification in Aerobic Chemostat Cultures of Thiosphaera pantotropha. AB - Thiosphaera pantotropha is capable of simultaneous heterotrophic nitrification and aerobic denitrification. Consequently, its nitrification potential could not be judged from nitrite accumulation, but was estimated from complete nitrogen balances. The maximum rate of nitrification obtained during these experiments was 93.9 nmol min mg of protein. The nitrification rate could be reduced by the provision of nitrate, nitrite, or thiosulfate to the culture medium. Both nitrification and denitrification increased as the dissolved oxygen concentration fell, until a critical level was reached at approximately 25% of air saturation. At this point, the rate of (aerobic) denitrification was equivalent to the anaerobic rate. At this dissolved oxygen concentration, the combined nitrification and denitrification was such that cultures receiving ammonium as their sole source of nitrogen appeared to become oxygen limited and the nitrification rate fell. It appeared that, under carbon-and energy-limited conditions, a high nitrification rate was correlated with a reduced biomass yield. To facilitate experimental design, a working hypothesis for the mechanism behind nitrification and denitrification by T. pantotropha was formulated. This involved the basic assumption that this species has a "bottleneck" in its cytochrome chain to oxygen and that denitrification and nitrification are used to overcome this. The nitrification potential of other heterotrophic nitrifiers has been reconsidered. Several species considered to be "poor" nitrifiers also simultaneously nitrify and denitrify, thus giving a falsely low nitrification potential. PMID- 16347781 TI - Analysis of Indole-3-Acetic Acid and Related Indoles in Culture Medium from Azospirillum lipoferum and Azospirillum brasilense. AB - Analysis of neutral and acidic ethyl acetate extracts from culture medium of Azospirillum brasilense 703Ebc by high-performance liquid chromatography (HPLC) and combined gas chromatography-mass spectrometry demonstrated the presence of indole-3-acetic acid (IAA), indole-3-ethanol, indole-3-methanol, and indole-3 lactic acid. IAA in media of 20 strains of A. brasilense and Azospirillum lipoferum was analyzed quantitatively by both the colorimetric Salkowski assay and HPLC-based isotopic dilution procedures. There was little correlation between the estimates obtained with the two procedures. For instance, the Salkowski assay suggested that the culture medium from A. brasilense 703Ebc contained 26.1 mug of IAA ml, whereas HPLC revealed the presence of only 0.5 mug of IAA ml. Equivalent estimates with A. brasilense 204Ed were 10.5 and 0.01 mug of IAA ml, respectively. The data demonstrate that the Salkowski assay is not a reliable method for measuring the IAA content of Azospirillum culture medium and that estimates in excess of 10 mug of IAA ml should be viewed with particular caution. Metabolism of [2'-C]IAA by A. brasilense 703Ebc yielded radiolabeled indole-3 methanol, whereas roots of maize (Zea mays L.) seedlings gave rise to [C]oxindole 3-acetic acid and an array of polar metabolites. Metabolism of [2'-C]IAA by maize roots inoculated with A. brasilense 703Ebc produced a metabolic profile characteristic of maize rather than Azospirillum species. PMID- 16347782 TI - Isolation of Rhizobium loti Strain-Specific DNA Sequences by Subtraction Hybridization. AB - Mixed-phase (heterogeneous) and single-phase (homogeneous) DNA subtraction hybridization methods were used to isolate specific DNA probes for closely related Rhizobium loti strains. In the heterogeneous method, DNA from the prospective probe strain was repeatedly hybridized to a mixture of DNA from cross hybridizing strains (subtracter DNA) which was immobilized on an epoxy-activated cellulose matrix. Probe strain sequences which shared homology with the matrix bound subtracter DNA hybridized to it, leaving unique probe strain sequences in the mobile phase. In the homogeneous method, probe strain sequences were hybridized in solution to biotinylated, mercurated subtracter DNA. Biotinylated, mercurated subtracer DNA and probe strain sequences hybridized to it were removed by two-step affinity chromatography on streptavidin-agarose and thiol-Sepharose. The specificity of the sequences remaining after subtraction hybridization by both methods was assessed and compared by colony hybridization with R. loti strains. Both methods allowed the rapid isolation of strain-specific DNA fragments which were suitable for use as probes. PMID- 16347783 TI - Improved Assay for Rhodanese in Thiobacillus spp. AB - Rhodanese (thiosulfate:cyanide sulfurtransferase; EC 2.8.1.1) catalyzes the conversion of thiosulfate and cyanide to thiocyanate and sulfite. Conventional rhodanese assays colorimetrically measure the formation of one or the other of the products. These assays suffer from the fact that there is significant nonbiological formation of these products in addition to the enzymatically catalyzed reaction. In the present report, we describe a modified procedure for assaying rhodanese in which a separate boiled control was prepared for each assay trial. The boiled control corrected for the nonbiological contributions to product formation. PMID- 16347784 TI - Isolation and Characterization of a Mutant of Arthrobacter sp. Strain GLP-1 Which Utilizes the Herbicide Glyphosate as Its Sole Source of Phosphorus and Nitrogen. AB - Arthrobacter sp. strain GLP-1, grown on glucose as a carbon source, utilizes the herbicide glyphosate [N-(phosphonomethyl)glycine] as its sole source of phosphorus as well as its sole source of nitrogen. The mutant strain GLP-1/Nit-1 utilizes glyphosate as its sole source of nitrogen as well. In strain GLP-1, P(i) was a potent competitive inhibitor of glyphosate uptake (K(i), 24 muM), while the affinity of P(i) for the uptake system of strain GLP-1/Nit-1 was reduced by 2 orders of magnitude (K(i), 2.3 mM). It is concluded that the inability of strain GLP-1 to utilize glyphosate as a source of nitrogen is due to the stringent control of glyphosate uptake by excess phosphate released during the degradation of the herbicide. PMID- 16347786 TI - Averaging of TNTC Counts. AB - [This corrects the article on p. 2069 in vol. 54.]. PMID- 16347785 TI - Purification and Characterization of Thermostable Pullulanase from Bacillus stearothermophilus and Molecular Cloning and Expression of the Gene in Bacillus subtilis. AB - A thermostable pullulanase (alpha-dextrin 6-glucanohydrolase [EC 3.2.1.41]) from a newly isolated Bacillus stearothermophilus strain (TRS128) was purified and characterized. The enzyme hydrolyzed (1-->6)-alpha-d-glucosidic linkages of pullulan to produce maltotriose, and the optimum temperature was 65 degrees C. About 90% of the enzyme activity was retained after treatment at 65 degrees C for 60 min. By using pTB522 as a vector plasmid, the pullulanase gene was cloned and expressed in Bacillus subtilis. PMID- 16347787 TI - Effects of Light and CO on the Survival of a Marine Ammonium-Oxidizing Bacterium during Energy Source Deprivation. AB - The chemolithotrophic ammonium-oxidizing bacterium Nitrosomonas cryotolerans responds uniquely to nutrient deprivation by lowering its endogenous respiration and anabolic processes to undetectable levels during starvation, thus appearing to enter a dormant state. To ascertain whether this state protects the cells from further stresses (as seen with endospore-forming bacteria), the starved cells were subjected to two known inhibitors, CO and light. It was found that long-term starved cells were less resistant than freshly starved cells to light inhibition. Both long-term-starved cells and freshly starved cells were unaffected by CO. PMID- 16347788 TI - Role of Cations in Accumulation and Release of Phosphate by Acinetobacter Strain 210A. AB - Cells of the strictly aerobic Acinetobacter strain 210A, containing aerobically large amounts of polyphosphate (100 mg of phosphorus per g [dry weight] of biomass), released in the absence of oxygen 1.49 mmol of P(i), 0.77 meq of Mg, 0.48 meq of K, 0.02 meq of Ca, and 0.14 meq of NH(4) per g (dry weight) of biomass. The drop in pH during this anaerobic phase was caused by the release of 1.8 protons per PO(4) molecule. Cells of Acinetobacter strain 132, which do not accumulate polyphosphate aerobically, released only 0.33 mmol of P(i) and 0.13 meq of Mg per g (dry weight) of biomass but released K in amounts comparable to those released by strain 210A. Stationary-phase cultures of Acinetobacter strain 210A, in which polyphosphate could not be detected by Neisser staining, aerobically took up phosphate simultaneously with Mg, the most important counterion in polyphosphate. In the absence of dissolved phosphate in the medium, no Mg was taken up. Cells containing polyphosphate granules were able to grow in a Mg-free medium, whereas cells without these granules were not. Mg was not essential as a counterion because it could be replaced by Ca. The presence of small amounts of K was essential for polyphosphate formation in cells of strain 210A. During continuous cultivation under K limitation, cells of Acinetobacter strain 210A contained only 14 mg of phosphorus per g (dry weight) of biomass, whereas this element was accumulated in amounts of 59 mg/g under substrate limitation and 41 mg/g under Mg limitation. For phosphate uptake in activated sludge, the presence of K seemed to be crucial. PMID- 16347789 TI - Plasmid incidence in bacteria from deep subsurface sediments. AB - Bacteria were isolated from deep terrestrial subsurface sediments underlying the coastal plain of South Carolina. A total of 163 isolates from deep sediments, surface soil, and return drill muds were examined for plasmid DNA content and resistance to the antibiotics penicillin, ampicillin, carbenicillin, streptomycin, kanamycin, and tetracycline. MICs of Cu, Cr, and Hg for each isolate were also determined. The overall frequency of plasmid occurrence in the subsurface bacteria was 33%. Resistance was most frequent to penicillin (70% of all isolates), ampicillin (49%), and carbenicillin (32%) and was concluded to be related to the concentrations of the individual antibiotics in the disks used for assaying resistance and to the production of low levels of beta-lactamase. The frequencies of resistance to penicillin and ampicillin were significantly greater for isolates bearing plasmids than for plasmidless isolates; however, resistance was not transferable to penicillin-sensitive Escherichia coli. Hybridization of subsurface bacterial plasmids and chromosomal DNA with a whole-TOL-plasmid (pWWO) probe revealed some homology of subsurface bacterial plasmid and chromosomal DNAs, indicating a potential for those bacteria to harbor catabolic genes on plasmids or chromosomes. The incidences of antibiotic resistance and MICs of metals for subsurface bacteria were significantly different from those for drill mud bacteria, ruling out the possibility that bacteria from sediments were derived from drill muds. PMID- 16347790 TI - Formation of Polyesters by Pseudomonas oleovorans: Effect of Substrates on Formation and Composition of Poly-(R)-3-Hydroxyalkanoates and Poly-(R)-3 Hydroxyalkenoates. AB - Pseudomonas oleovorans grows on C(6) to C(12)n-alkanes and 1-alkenes. These substrates are oxidized to the corresponding fatty acids, which are oxidized further via the beta-oxidation pathway, yielding shorter fatty acids which have lost one or more C(2) units. P. oleovorans normally utilizes beta-oxidation pathway intermediates for growth, but in this paper we show that the intermediate 3-hydroxy fatty acids can also be polymerized to intracellular poly-(R)-3 hydroxyalkanoates (PHAs) when the medium contains limiting amounts of essential elements, such as nitrogen. The monomer composition of these polyesters is a reflection of the substrates used for growth of P. oleovorans. The largest monomer found in PHAs always contained as many C atoms as did the n-alkane used as a substrate. Monomers which were shorter by one or more C(2) units were also observed. Thus, for C-even substrates, only C-even monomers were found, the smallest being (R)-3-hydroxyhexanoate. For C-odd substrates, only C-odd monomers were found, with (R)-3-hydroxyheptanoate as the smallest monomer. 1-Alkenes were also incorporated into PHAs, albeit less efficiently and with lower yields than n alkanes. These PHAs contained both saturated and unsaturated monomers, apparently because the 1-alkene substrates could be oxidized to carboxylic acids at either the saturated or the unsaturated ends. Up to 55% of the PHA monomers contained terminal double bonds when P. oleovorans was grown on 1-alkenes. The degree of unsaturation of PHAs could be modulated by varying the ratio of alkenes to alkanes in the growth medium. Since 1-alkenes were also shortened before being polymerized, as was the case for n-alkanes, copolymers which varied with respect to both monomer chain length and the percentage of terminal double bonds were formed during nitrogen-limited growth of P. oleovorans on 1-alkenes. Such polymers are expected to be useful for future chemical modifications. PMID- 16347791 TI - Presence of an unusual methanogenic bacterium in coal gasification waste. AB - Methanogenic bacteria growing on a pilot-scale, anaerobic filter processing coal gasification waste were enriched in a mineral salts medium containing hydrogen and acetate as potential energy sources. Transfer of the enrichments to methanol medium resulted in the initial growth of a strain of Methanosarcina barkeri, but eventually small cocci became dominant. The cocci growing on methanol produced methane and exhibited the typical fluorescence of methanogenic bacteria. They grew in the presence of the cell wall synthesis-inhibiting antibiotics d cycloserine, fosfomycin, penicillin G, and vancomycin as well as in the presence of kanamycin, an inhibitor of protein synthesis in eubacteria. The optimal growth temperature was 37 degrees C, and the doubling time was 7.5 h. The strain lysed after reaching stationary phase. The bacterium grew poorly with hydrogen as the energy source and failed to grow on acetate. Morphologically, the coccus shared similarities with Methanosarcina sp. Cells were 1 mum wide, exhibited the typical thick cell wall and cross-wall formation, and formed tetrads. Packets and cysts were not formed. PMID- 16347792 TI - Floc Formation by Azospirillum lipoferum Grown on Poly-beta-Hydroxybutyrate. AB - Azospirillum lipoferum RG6xx was grown under conditions similar to those resulting in encystment of Azotobacter spp. A. lipoferum produced cells of uniform shape when grown on nitrogen-free beta-hydroxybutyrate agar. Cells accumulated poly-beta-hydroxybutyrate and often grew as chains or filaments that eventually lost motility and formed capsules. Within 1 week, vegetative A. lipoferum inocula were converted into microflocs arising from filaments or chains. Cells within microflocs were pleomorphic, contained much poly-beta hydroxybutyrate, and were encapsulated. Some cells had a cystlike morphology. Up to 57% of the dry weight of encapsulated flocs was poly-beta-hydroxybutyrate, whereas vegetative cells grown in broth with combined nitrogen had only 3% of their dry weight as poly-beta-hydroxybutyrate. Neither encapsulated cells in flocs nor nonencapsulated vegetative cells were significantly desiccation resistant. Under starvation conditions (9 days) only 25% of encapsulated cells remained viable, whereas vegetative cells multiplied severalfold. In short-term germination experiments with encapsulated flocs, nitrate, ammonium, and soil extract promoted formation of motile vegetative cells. Most cells in treatments lacking combined nitrogen eventually depleted their visible poly-beta hydroxybutyrate reserves without germinating. The remaining cells retained the reserve polymer and underwent size reduction. PMID- 16347793 TI - Identification and Characterization of a Bacteroid-Specific Dehydrogenase Complex in Rhizobium leguminosarum PRE. AB - In membranes of Rhizobium leguminosarum bacteroids isolated from nitrogen-fixing pea root nodules, two different protein complexes with NADH dehydrogenase activity were detected. One of these complexes, with a molecular mass of 110 kilodaltons, was also found in membranes of free-living rhizobia, but the other, with a molecular mass of 550 kilodaltons, appeared to be present only in bacteroids. The bacteroid-specific complex, referred to as DH1, probably consists of at least four different subunits. Using antibodies raised against the separate polypeptides, we found that a 35,000-molecular-weight polypeptide (35K polypeptide) in the DH1 complex is bacteroid specific, while the other proposed subunits were also detectable in cytoplasmic membranes of free-living bacteria. Dehydrogenase complex DH1 is also present in bacteroids of a R. leguminosarum nifA mutant, indicating that the synthesis of the dehydrogenase is not dependent on the gene product of this nif-regulatory gene. A possible involvement of the bacteroid-specific DH1 complex in electron transport to nitrogenase is discussed. PMID- 16347794 TI - Pressure and Temperature Effects on Growth and Methane Production of the Extreme Thermophile Methanococcus jannaschii. AB - The marine archaebacterium Methanococcus jannaschii was studied at high temperatures and hyperbaric pressures of helium to investigate the effect of pressure on the behavior of a deep-sea thermophile. Methanogenesis and growth (as measured by protein production) at both 86 and 90 degrees C were accelerated by pressure up to 750 atm (1 atm = 101.29kPa), but growth was not observed above 90 degrees C at either 7.8 or 250 atm. However, growth and methanogenesis were uncoupled above 90 degrees C, and the high-temperature limit for methanogenesis was increased by pressure. Substantial methane formation was evident at 98 degrees C and 250 atm, whereas no methane formation was observed at 94 degrees C and 7.8 atm. In contrast, when argon was substituted for helium as the pressurizing gas at 250 atm, no methane was produced at 86 degrees C. Methanogenesis was also suppressed at 86 degrees C and 250 atm when the culture was pressurized with a 4:1 mix of H(2) and CO(2), although limited methanogenesis did occur when the culture was pressurized with H(2). PMID- 16347795 TI - Production of succinic Acid from citric Acid and related acids by lactobacillus strains. AB - A number of Lactobacillus strains produced succinic acid in de Man-Rogosa-Sharpe broth to various extents. Among 86 fresh isolates from fermented cane molasses in Thailand, 30 strains (35%) produced succinic acid; namely, 23 of 39 Lactobacillus reuteri strains, 6 of 18 L. cellobiosus strains, and 1 of 6 unidentified strains. All of 10 L. casei subsp. casei strains, 5 L. casei subsp. rhamnosus strains, 6 L. mali strains, and 2 L. buchneri strains did not produce succinic acid. Among 58 known strains including 48 type strains of different Lactobacillus species, the strains of L. acidophilus, L. crispatus, L. jensenii, and L. parvus produced succinic acid to the same extent as the most active fresh isolates, and those of L. alimentarius, L. collinoides, L. farciminis, L. fructivorans (1 of 2 strains tested), L. malefermentans, and L. reuteri were also positive, to lesser extents. Diammonium citrate in de Man-Rogosa-Sharpe broth was determined as a precursor of the succinic acid produced. Production rates were about 70% on a molar basis with two fresh strains tested. Succinic acid was also produced from fumaric and malic acids but not from dl-isocitric, alpha-ketoglutaric, and pyruvic acids. The present study is considered to provide the first evidence on the production of succinic acid, an important flavoring substance in dairy products and fermented beverages, from citrate by lactobacilli. PMID- 16347796 TI - Degradation of Kraft Indulin Lignin by Streptomyces viridosporus and Streptomyces badius. AB - Crawford and collaborators have studied extensively the solubilization of lignocellulose by two Streptomyces species, S. badius and S. viridosporus. Using a condensed industrial lignin essentially devoid of carbohydrates, Indulin AT, as the sole source of carbon, similar results were obtained: (i) the growths of the bacteria were optimum at pH 7.5 to 8.5; (ii) yeast extract was a better source of nitrogen than NH(4)Cl; (iii) the products of the depolymerization of Indulin were soluble, acid-precipitable polymers. When d-glucose was added as a secondary carbon source, it was used preferentially and the production of acid-precipitable polymers began only after the complete depletion of the sugar. On the assumption that the degradation of Indulin was catalyzed by enzymes, proteins found in the culture media and soluble and insoluble intracellular proteins were incubated with Indulin at pH 7.0 at 37 degrees C. Proteins in all fractions from S. badius had ligninolytic activities which, with the exception of those in the intracellular soluble fraction, were increased in the presence of H(2)O(2). In S. viridosporus, both extra- and intracellular soluble activities were found which were not increased by H(2)O(2). The extracellular activity of S. viridosporus was not affected by heat, resisted partially an exposure to pH 1.0, and was completely destroyed by proteolysis. PMID- 16347797 TI - Examination of Genetic Relatedness of Marine Synechococcus spp. by Using Restriction Fragment Length Polymorphisms. AB - The relatedness of several marine Synechococcus spp. was estimated by DNA hybridization. Strains isolated from various geographical locations and representing a diversity of DNA base compositions and phycobiliprotein profiles were compared by restriction fragment length polymorphisms for a number of genes. DNAs from two marine red algae and a cryptomonad alga (which exhibit a phycobiliprotein composition similar to that of the marine Synechococcus spp.) and Synechococcus strain PCC6301 (Anacystis nidulans) were also included in the comparison. Strains WH8008, WH8018, and WH7805 were shown to be very similar to one another, as were strains WH7802 and WH7803. Strains WH8110 and WH5701 were clearly unrelated to any of the other strains, and no marine Synechococcus isolate showed any similarity to the freshwater Synechococcus strain PCC6301 or the eucaryotic algae. The method is relatively straightforward and sensitive and uses a variety of basic molecular biology techniques. Its utility in ascertaining the genetic relatedness and diversity of marine Synechococcus spp. and possible extension to field studies are discussed. PMID- 16347798 TI - Fast kinetics of fe oxidation in packed-bed reactors. AB - Thiobacillus ferrooxidans was used in fixed-film bioreactors to oxidize ferrous sulfate to ferric sulfate. Glass beads, ion-exchange resin, and activated-carbon particles were tested as support matrix materials. Activated carbon was tested in both a packed-bed bioreactor and a fluidized-bed bioreactor; the other matrix materials were used in packed-bed reactors. Activated carbon displayed the most suitable characteristics for use as a support matrix of T. ferrooxidans fixed film formation. The reactors were operated within a pH range of 1.35 to 1.5, which effectively reduced the amount of ferric iron precipitation and eliminated diffusion control of mass transfer due to precipitation. The activated-carbon packed-bed reactor displayed the most favorable biomass holdup and kinetic performance related to ferrous sulfate oxidation. The fastest kinetic performance achieved with the activated-carbon packed-bed bioreactor was 78 g of Fe oxidized per liter per h (1,400 mmol of Fe oxidized per liter per h) at a true dilution rate of 40/h, which represents a hydraulic retention time of 1.5 min. PMID- 16347799 TI - Characterization of Jarosite Formed upon Bacterial Oxidation of Ferrous Sulfate in a Packed-Bed Reactor. AB - A packed-bed bioreactor with activated-carbon particles as a carrier matrix material inoculated with Thiobacillus ferrooxidans was operated at a pH of 1.35 to 1.5 to convert ferrous sulfate to ferric sulfate. Despite the low operating pH, trace amounts of precipitates were produced in both the reactor and the oxidized effluent. X-ray diffraction and chemical analyses indicated that the precipitates were well-ordered potassium jarosite. The chemical analyses also revealed a relative deficiency of Fe and an excess of S in the reactor sample compared with the theoretical composition of potassium jarosite. PMID- 16347800 TI - Quantitative assessment of factors affecting the recovery of indigenous and released thermophilic bacteria from compost. AB - Thermophilic actinomycetes and bacilli were recovered from mushroom compost by conventional dilution plating and sedimentation chamber-Andersen sampler methods. Excessive growth of thermophilic bacilli on dilution plates accounted for the poor recovery and limited diversity of actinomycete colonies, and this result was largely unaffected by the use of modified extraction procedures and diluents. Assessment of the actinomycete population was more successfully achieved by applying the sedimentation chamber method, by using selective media, or both. Background resistance of the compost microflora to selective agents (kanamycin, novobiocin, tetracycline, thiostrepton, and NaCl) was extremely varied, but both actinomycetes and bacilli were particularly sensitive to tetracycline. The selective isolation of Thermoactinomyces spp. and Thermomonospora chromogena by novobiocin and kanamycin, respectively, was shown to be reproducible, and the use of high concentrations of kanamycin resulted in the isolation of a novel group of unidentified thermophilic actinomycetes. Comparison of nonselective nutrient media demonstrated that the nutrient-rich protoplast regeneration medium R5 was surprisingly efficient for actinomycete recovery. This medium was found to be particularly appropriate for the recovery of Saccharomonospora viridis BD125, introduced as spores into both sterile and fresh samples of mushroom compost. This stable pigmented variant of the S. viridis strains indigenous to compost was released at concentrations of up to 10 spores g of compost in order to provide information for future experiments on the release and recovery of genetically manipulated strains. The detection limits for this strain were in the region of 10 g from sterilized compost but only 10 g from nonsterile compost. These figures correspond to mean recovery efficiencies of approximately 70% (sterilized compost) and 53% (fresh compost) of viable spores released. Further improvements in the detection and recovery of S. viridis strains released into compost should be achieved by the introduction of selectable markers developed from this information on the antibiotic resistance profile of the indigenous compost microflora. PMID- 16347801 TI - Protozoan grazing, bacterial activity, and mineralization in two-stage continuous cultures. AB - In two-stage continuous cultures, at bacterial concentrations, biovolumes, and growth rates similar to values found in Lake Vechten, ingestion rates of heterotrophic nanoflagellates (HNAN) increased from 2.3 bacteria HNAN . h at a growth rate of 0.15 day to 9.2 bacteria . HNAN . h at a growth rate of 0.65 day. On a yeast extract medium with a C/N/P ratio of 100:15:1.2 (Redfield ratio), a mixed bacterial population showed a yield of 18% (C/C) and a specific carbon content of 211 fg of C . mum. The HNAN carbon content and yield were estimated at 127 fg of C . mum and 47% (C/C). Although P was not growth limiting, HNAN accelerated the mineralization of PO(4)-P from dissolved organic matter by 600%. The major mechanism of P remineralization appeared to be direct consumption of bacteria by HNAN. N mineralization was performed mainly (70%) by bacteria but was increased 30% by HNAN. HNAN did not enhance the decomposition of the relatively mineral-rich dissolved organic matter. An accelerated decomposition of organic carbon by protozoa may be restricted to mineral-poor substrates and may be explained mainly by protozoan nutrient regeneration. Growth and grazing in the cultures were compared with methods for in situ estimates. Thymidine incorporation by actively growing bacteria yielded an empirical conversion factor of 1.1 x 10 bacteria per mol of thymidine incorporated into DNA. However, nongrowing bacteria also showed considerable incorporation. Protozoan grazing was found to be accurately measured by uptake of fluorescently labeled bacteria, whereas artificial fluorescent microspheres were not ingested, and selective prokaryotic inhibitors blocked not only bacterial growth but also protozoan grazing. PMID- 16347802 TI - Colonization of Tomato Plants by Two Agrocin-Producing Strains of Agrobacterium tumefaciens. AB - For a bacterium to be a successful biocontrol agent against crown gall disease, it must produce an effective agrocin specific for Agrobacterium tumefaciens and be able to colonize host plants efficiently. The colonization abilities of K84 and J73, successful and potential biocontrolling strains, respectively, were compared both in vivo and in vitro. Both strains produced fibrils attaching them to tomato root surfaces and had similar colonization efficiencies up to 14 days after inoculation. However, the ability of J73 to colonize plants for longer periods was significantly less than that of K84. Thus, the presence of fibrils is not sufficient to ensure colonization. No correlation was found between hydrophobicity and colonization. PMID- 16347803 TI - Induction of Yellow Pigmentation in Serratia marcescens. AB - The appearance of yellow pigmentation in nonpigmented strains of Serratia sp. has been demonstrated to be due to the production of a muconic acid, 2-hydroxy-5 carboxymethylmuconic acid semialdehyde. The 3,4-dihydroxyphenylacetate 2,3 dioxygenase responsible for the synthesis of this muconic acid was induced in all strains tested. Another muconic acid, the beta-cis-cis-carboxymuconic acid, could also be synthesized from 3,4-dihydroxybenzoate, but this product was not colored. Mutants that were unable to grow on tyrosine and produced yellow pigment were isolated from nonpigmented strains. These mutants had properties similar to those of the yellow-pigmented strains. The ability to produce pigment may be more widespread among Serratia marcescens strains than is currently known. PMID- 16347804 TI - Production and further characterization of an alkaline elastase produced by alkalophilic bacillus strain ya-B. AB - The characteristics of the obligate alkalophilic Bacillus sp. strain Ya-B, which produces alkaline elastase extracellularly, were examined. This strain grew at pH 7.0 only in the presence of 1% or more NaCl. Its fatty acid distribution pattern was similar to that of other Bacillus species in which iso-C(15) and anteiso C(15) were the most abundant fatty acids. About 120 mg of enzyme was recovered from 1 liter of culture broth in a medium (pH 10.1) containing mainly glucose, soymeal, and glycerol. The antiserum against this enzyme did not recognize microbial proteinases, such as subtilisins, but reacted with proteinase C, which was purified from commercial pronase. Chemical modification studies revealed that certain histidine and tyrosine residues might be involved in the enzyme activity. This enzyme underwent a partial unfolding at pHs higher than 12.0, as indicated by the circular dichroism study. PMID- 16347805 TI - Useful Host-Vector Systems in Bacillus stearothermophilus. AB - We isolated a highly transformable thermophile, Bacillus stearothermophilus SIC1, which exhibited the following features. The growth temperature ranged from 45 to 65 degrees C in L broth. The maximum cell concentration in 2L broth (2% tryptone, 1% yeast extract, 0.5% NaCl, pH 7.2) was determined as an optical density at 660 nm of 7.8, and the generation time was 11 min at 60 degrees C. Strain SIC1 was a prototroph and was transformed by the protoplast procedure not only with repB plasmids (high-copy-number plasmids such as pTB913 and pUB110) but also with repA plasmids (low-copy-number plasmids such as pTB53). Transformation efficiencies with repB and repA plasmids were about 2 x 10 to 5 x 10 and 5 x 10 transformants per mug of DNA, respectively. The transformant carrying plasmid pTB913Y/K could grow at 63 degrees C in the presence of kanamycin. The regeneration frequency of protoplasts was 60%, and only 1 day was needed for regeneration at 55 degrees C. PMID- 16347806 TI - Underestimation of DNA synthesis by [h]thymidine incorporation in marine bacteria. AB - A direct comparison of [H]thymidine incorporation with DNA synthesis was made by using an exponentially growing estuarine bacterial isolate and the naturally occurring bacterial populations in a eutrophic subtropical estuary and in oligotrophic offshore waters. Simultaneous measurements of [H]thymidine incorporation into DNA, fluorometrically determined DNA content, and direct counts were made over time. DNA synthesis estimated from thymidine incorporation values was compared with fluorometrically determined changes in DNA content. Even after isotope dilution, nonspecific macromolecular labeling, and efficiency of DNA recovery were accounted for, [H]thymidine incorporation consistently underestimated DNA synthesized by six- to eightfold. These results indicate that although the relationship of [H]thymidine incorporation to DNA synthesis appears consistent, there are significant sources of thymine bases incorporated into DNA which cannot be accounted for by standard [H]thymidine incorporation and isotope dilution assays. PMID- 16347807 TI - Tn5-Mediated Cloning of a Genetic Region from Pseudomonas putida Involved in the Stimulation of Plant Root Elongation. AB - Transposon (Tn5) mutagenesis was applied to Pseudomonas putida GR12-2R3, which promotes root elongation (a phenotype designated Pre) of Brassica campestris under gnotobiotic conditions. Of 3,000 Tn5 transconjugants, only one mutant that lost Pre activity but remained prototrophic and capable of plant root colonization was detected. This mutant was complemented by plasmid pRE53, which contained a 15.0-kilobase DNA insert isolated from a parental strain. The complemented mutant regained full Pre activity comparable to that of the wild type. PMID- 16347808 TI - Transfer and expression of lithoautotrophy and denitrification in a host lacking these metabolic activities. AB - The conjugative 450-kilobase-pair megaplasmid pHG1 from Alcaligenes eutrophus H16 was transferred to the herbicide-degrading soil bacterium A. eutrophus JMP134. This transfer was achieved by means of RP4 mobilization and a Tn5-Mob insertion provided in trans on the megaplasmid replicon. Although kanamycin-resistant transconjugants also occurred with other gram-negative species such as Rhizobium, Agrobacterium, and thiobacteria, A. eutrophus JMP134 was the only recipient which stably maintained the megaplasmid. pHG1-containing transconjugants derived from JMP134 expressed all metabolic functions associated with the plasmid: the ability to oxidize hydrogen through catalysis of two hydrogenases, to assimilate carbon dioxide via the Calvin cycle pathway, and to grow with nitrate anaerobically. All of these metabolic activities were absent in the original strain JMP134. PMID- 16347809 TI - Disk inoculum-solid medium method to test carbon and nitrogen assimilation by yeast isolates. AB - Carbon and nitrogen assimilation for 50 yeasts isolated from White Riesling fermentation were tested by using a disk inoculum-solid medium method. This method was quicker and gave results comparable to the conventional liquid medium methods. Yeast characteristics (growth response, pigment production, morphology) could also be compared with this method. PMID- 16347810 TI - Interaction of Ammonia Monooxygenase from Nitrosomonas europaea with Alkanes, Alkenes, and Alkynes. AB - Ammonia monooxygenase of Nitrosomonas europaea catalyzes the oxidation of alkanes (up to C(8)) to alcohols and alkenes (up to C(5)) to epoxides and alcohols in the presence of ammonium ions. Straight-chain, N-terminal alkynes (up to C(10)) all exhibited a time-dependent inhibition of ammonia oxidation without effects on hydrazine oxidation. PMID- 16347811 TI - Ethanol extraction requirement for purification of protein labeled with [h]leucine in aquatic bacterial production studies. AB - The trichloroacetic acid (TCA)-insoluble fraction of water column bacteria labeled with [H]leucine contained an ethanol-soluble fraction accounting for up to 44% of the label. A component of the ethanol-soluble fraction is [H]leucine. Labeled-protein purification requires an ethanol wash step. Cold TCA can replace hot TCA for precipitation of labeled proteins. PMID- 16347812 TI - Control of Lignin Peroxidase Production by Phanerochaete chrysosporium INA-12 by Temperature Shifting. AB - There are two temperature optima connected with lignin peroxidase synthesis by Phanerochaete chrysosporium INA-12. One, at 37 degrees C, is for the mycelium growing phase; the other, at 30 degrees C, is for the lignin peroxidase-producing phase. One of six extracellular proteins with ligninase activity increased when cultures were grown at 30 degrees C for the entire fermentation period or when cultures were grown at 37 degrees C for the first 2 days of incubation and then shifted to 30 degrees C, compared with the activity of control cultures grown at 37 degrees C for the entire fermentation period. The unsaturation of fatty acid (Delta/mole) of P. chrysosporium INA-12 mycelium decreased from 1.25 to 1.03 when the growth temperature was shifted from 20 to 40 degrees C. PMID- 16347813 TI - National Field Evaluation of a Defined Substrate Method for the Simultaneous Enumeration of Total Coliforms and Escherichia coli from Drinking Water: Comparison with the Standard Multiple Tube Fermentation Method. AB - [This corrects the article on p. 1596 in vol. 54.]. PMID- 16347814 TI - Diversity within Serogroups of Rhizobium leguminosarum biovar viceae in the Palouse Region of Eastern Washington as Indicated by Plasmid Profiles, Intrinsic Antibiotic Resistance, and Topography. AB - Serology, plasmid profiles, and intrinsic antibiotic resistance (IAR) were determined for 192 isolates of Rhizobium leguminosarum biovar viceae from nodules of peas (Pisum sativum L.) grown on the south slope and bottomland topographic positions in eastern Washington State. A total of 3 serogroups and 18 plasmid profile groups were identified. Nearly all isolates within each plasmid profile group were specific for one of the three serogroups. Cluster analysis of IAR data showed that individual clusters were dominated by one serogroup and by one or two plasmid profile groups. Plasmid profile analysis and IAR analysis grouped 72% of the isolates similarly. Most plasmid profile groups and several IAR clusters favored either the south slope or the bottomland topographic position. These findings show that certain intraserogroup strains possess a greater competitiveness for nodulation and/or possess a greater ability to survive in adjacent soil environments. PMID- 16347815 TI - Isolation of Phaffia rhodozyma Mutants with Increased Astaxanthin Content. AB - Plating of the astaxanthin-producing yeast Phaffia rhodozyma onto yeast-malt agar containing 50 muM antimycin A gave rise to colonies of unusual morphology, characterized by a nonpigmented lower smooth surface that developed highly pigmented vertical papillae after 1 to 2 months. Isolation and purification of the pigmented papillae, followed by testing for pigment production in shake flasks, demonstrated that several antimycin isolates were increased two- to fivefold in astaxanthin content compared with the parental natural isolate (UCD FST 67-385). One of the antimycin strains (ant-1) and a nitrosoguanidine derivative of ant-1 (ant-1-4) produced considerably more astaxanthin than the parent (ant-1 had 800 to 900 mug/g; ant-1-4 had 900 to 1,300 mug/g; and 67-385 had 300 to 450 mug/g). The mutant strains were compared physiologically with the parent. The antimycin mutants grew slower on ammonia, glutamate, or glutamine as nitrogen sources compared with the natural isolate and also had lower cell yields on several carbon sources. Although isolated on antimycin plates, they were found to be more susceptible to antimycin A, apparently owing to the spatial separation of the papillae from the agar. They were also more susceptible than the parent to the respiratory inhibitor thenoyltrifluoroacetone and were slightly more susceptible to cyanide, but did not differ from the natural isolate in susceptibility to azide. The antimycin-derived strains were also killed faster than the parent by hydrogen peroxide. The carotenoid compositions of the parent and the antimycin-derived strains were similar to those previously determined in the type strain (UCD-FST 67-210) except that two carotenoids not previously found in the type strain were present in increased quantities in the antimycin mutants and phoenicoxanthin was a minor component. The chemical properties of the unknown carotenoids suggested that the strains isolated on antimycin agar tended to oxygenate and desaturate carotene precursors to a greater extent than the parent. The physiology of the antimycin isolates and the known specificity of antimycin for cytochrome b in the respiratory chain suggests that alteration of cytochrome b or cytochrome P-450 components involved in oxygenation and desaturation of carotenes in mitochondria are affected, which results in increased astaxanthin production. These astaxanthin-overproducing mutants and more highly pigmented derivative strains could be useful in providing a natural source of astaxanthin for the pen-reared-salmon industry or for other farmed animals that contain astaxanthin as their principal carotenoid. PMID- 16347816 TI - Characteristics of Hormogonia Formation by Symbiotic Nostoc spp. in Response to the Presence of Anthoceros punctatus or Its Extracellular Products. AB - Nostocacean cyanobacteria typically produce gliding filaments termed hormogonia at a low frequency as part of their life cycle. We report here that all Nostoc spp. competent in establishing a symbiotic association with the hornwort Anthoceros punctatus formed hormogonial filaments at a high frequency in the presence of A. punctatus. The hormogonia-inducing activity was produced by A. punctatus under nitrogen-limited culture conditions. The hormogonia of the symbiotically competent Nostoc spp. were characterized as motile (gliding) filaments lacking heterocysts and with distinctly smaller cells than those of vegetative filaments; the small cells resulted from a continuation of cell division uncoupled from biomass increase. An essentially complete conversion of vegetative filaments to hormogonia occurred within 12 h of exposure of Nostoc sp. strain 7801 to A. punctatus growth-conditioned medium. Hormogonia formation was accompanied by loss of nitrogen fixation (acetylene reduction) and by decreases in photosynthetic CO(2) fixation and in vivo NH(4) assimilation of 30% and approximately 40%, respectively. The rates of acetylene reduction and CO(2) fixation returned to approximately the control rates within 72 to 96 h after hormogonia induction, as the cultures of Nostoc sp. strain 7801 differentiated heterocysts and reverted to the vegetative growth state. The relationship between hormogonia formation and symbiotic competence is discussed. PMID- 16347817 TI - Characterization of Xylose Uptake in the Yeasts Pichia heedii and Pichia stipitis. AB - The kinetics of xylose uptake were investigated in the efficient xylose fermenter Pichia stipitis and in the more readily genetically manipulated, strictly respiratory yeast Pichia heedii. Both yeasts demonstrated more than one xylose uptake system, differing in substrate affinity. The K(m) of high-affinity xylose uptake in both organisms was similar to that of the efficient high-affinity glucose uptake system of Saccharomyces cerevisiae. In P. heedii, low-affinity xylose uptake was enhanced with growth on 2% but not 0.05% xylose and high affinity uptake was reduced. In contrast to glucose uptake, xylose uptake in P. heedii was inhibited by dinitrophenol. Dinitrophenol inhibited both glucose and xylose uptake by P. stipitis. Glucose uptake was not inhibited by a 100-fold molar excess of xylose in P. heedii. It is suggested that xylose uptake in P. heedii is via a carrier system(s) distinct from those for glucose uptake. PMID- 16347818 TI - Acetate-Activating Enzymes of Bradyrhizobium japonicum Bacteroids. AB - Acetyl coenzyme A (acetyl-CoA) synthetase and acetate kinase were localized within the soluble portion of Bradyrhizobium japonicum bacteroids, and no appreciable activity was found elsewhere in the nodule. The presence of each acetate-activating enzyme was confirmed by separation of the two enzyme activities on a hydroxylapatite column, by substrate dependence of each enzyme in both the forward and reverse directions, by substrate specificity, by inhibition patterns, and also by identification of the reaction products by C(18) reverse phase high-pressure liquid chromatography. Phosphotransacetylase activity, found in the soluble portion of the bacteroid, was dependent on the presence of potassium and was inhibited by added sodium. The greatest acetyl-CoA hydrolase activity was found in the root nodule cytosol, although appreciable activity also was found within the bacteroids. The combined specific activities of acetyl-CoA synthetase and acetate kinase-phosphotransacetylase were approximate to that of the pyruvate dehydrogenase complex, thus providing B. japonicum with sufficient capacity to generate acetyl-CoA. PMID- 16347819 TI - Continuous and Batch Production of Chloroperoxidase by Mycelial Pellets of Caldariomyces fumago in an Airlift Fermentor. AB - Batch and continuous production of the extracellular heme glycoprotein chloroperoxidase (CPO) was studied with an airlift fermentor. We induced Caldariomyces fumago CMI 89362 to form pellets by transferring a small inoculum volume in preculture prior to growth in a 1-liter fermentor. Continuous replacement of the fructose-salts medium (dilution rate, 0.008 h) supported continuous CPO formation at an average concentration of 128 +/- 10 mg of CPO liter for 8 days. Optimum CPO production rates averaged 1.2 +/- 0.1 mg of CPO h at dilution rates below 0.033 h. Varying the carbohydrate content of the feed solution or the time of starting the feed did not significantly alter the amount of CPO produced. Batch fermentation in the airlift fermentor resulted in maximum CPO concentrations of 280 +/- 80 mg of CPO liter, although on two separate occasions CPO concentrations reached 400 to 450 mg liter, which was double the amount obtained by free hyphae in shake flask culture. PMID- 16347820 TI - Cloning of the Membrane-Bound Aldehyde Dehydrogenase Gene of Acetobacter polyoxogenes and Improvement of Acetic Acid Production by Use of the Cloned Gene. AB - A genomic clone bank of Acetobacter polyoxogenes NBI1028 constructed in Escherichia coli by use of the expression vector pUC18 was screened with antibody raised against membrane-bound aldehyde dehydrogenase (ALDH; 75 kilodaltons [kDa]) from A. polyoxogenes NBI1028. A clone that synthesized a 41-kDa protein cross reactive with anti-ALDH antibody was isolated. For cloning of the full-length ALDH structural gene, a cosmid gene bank was screened by Southern blot hybridization with the cloned DNA as a probe, and subcloning from the positive cosmid clone was performed with shuttle vector pMV24. Plasmid pAL25, containing the full-length ALDH structural gene, was isolated and expressed in both E. coli and Acetobacter aceti to produce a fused protein (78 kDa) with a short NH(2) terminal beta-galactosidase peptide. pAL25 conferred ALDH production on a mutant of A. aceti lacking the enzyme activity. Transformation of A. aceti subsp. xylinum NBI2099 with pAL25 caused 2- and 1.4-fold increases in the production rate and in the maximum concentration of acetic acid in submerged fermentation, respectively. PMID- 16347821 TI - Purification and Characterization of an Endo-(1,3)-beta-d-Glucanase from Trichoderma longibrachiatum. AB - A laminarinase [endo-(1,3)-beta-d-glucanase] has been purified from Trichoderma longibrachiatum cultivated with d-glucose as the growth substrate. The enzyme was found to hydrolyze laminarin to oligosaccharides varying in size from glucose to pentaose and to lesser amounts of larger oligosaccharides. The enzyme was unable to cleave laminaribiose but hydrolyzed triose to laminaribiose and glucose. The enzyme cleaved laminaritetraose, yielding laminaritriose, laminaribiose, and glucose, and similarly cleaved laminaripentaose, yielding laminaritetraose, laminaritriose, laminaribiose, and glucose. The enzyme cleaved only glucans containing beta-1,3 linkages. The pH and temperature optima were 4.8 and 55 degrees C, respectively. Stability in the absence of a substrate was observed at temperatures up to 50 degrees C and at pH values between 4.9 and 9.3. The molecular mass was determined to be 70 kilodaltons by sodium dodecyl sulfate 12.5% polyacrylamide gel electrophoresis, and the pI was 7.2. Enzyme activity was significantly inhibited in the presence of HgCl(2), MnCl(2), KMnO(4), and N bromosuccinimide. The K(m) of the enzyme on laminarin was 0.0016%, and the V(max) on laminarin was 3,170 mumol of glucose equivalents per mg of the pure enzyme per min. PMID- 16347822 TI - Sugar Utilization and Acid Production by Free and Entrapped Cells of Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis subsp. lactis in a Whey Permeate Medium. AB - Cells of Streptococcus salivarius subsp. thermophilus and Lactococcus lactis subsp. lactis entrapped in k-carrageenan-locust bean gum gel performed similarly to free cells in the conversion of lactose to lactic acid. Bead diameter influenced the fermentation rate. Cells entrapped in smaller beads (0.5 to 1.0 mm) showed higher release rates, higher lactose, glucose, and formic acid utilization, higher galactose accumulation, and higher lactic acid production than did cells entrapped in larger beads (1.0 to 2.0 mm). Values for smaller beads were comparable with those for free cells. Immobilization affected the fermentation rate of lactic acid bacteria, especially Lactobacillus delbrueckii subsp. bulgaricus. Entrapped cells of L. delbrueckii subsp. bulgaricus demonstrated a lower lactic acid production than did free cells in batch fermentation. The kinetics of the production of formic and pyruvic acids by L. lactis subsp. lactis and S. salivarius subsp. thermophilus are presented. PMID- 16347823 TI - Intact soil-core microcosms for evaluating the fate and ecological impact of the release of genetically engineered microorganisms. AB - Intact soil-core microcosms were studied to determine their applicability for evaluating the transport, survival, and potential ecosystem effects of genetically engineered microorganisms before they are released into the environment. Soil-core microcosms were planted with wheat and maize seeds and inoculated with Azospirillum lipoferum SpBr17 and SpRG20a Tn5 mutants, respectively. Microcosm leachate, rhizosphere soil, plant endorhizosphere, insects, and xylem exudate were sampled for A. lipoferum Tn5 mutant populations. A. lipoferum Tn5 populations, determined by most-probable-number technique-DNA hybridization, varied from below detection to 10 g of dry root in the rhizosphere, with smaller populations detected in the endorhizosphere. Intact soil-core microcosms were found to maintain some of the complexities of the natural ecosystem and should be particularly useful for initial evaluations of the fate of plant-associated genetically engineered bacteria. PMID- 16347824 TI - Cellulose Fermentation by an Asporogenous Mutant and an Ethanol-Tolerant Mutant of Clostridium thermocellum. AB - Two mutants of Clostridium thermocellum were isolated after UV light mutagenesis. Mutant A1, selected as asporogenous, exhibited a fermentation pattern similar to that of the wild type. However, at pH 6.5, the mutant degraded 12% more cellulose than did the wild type, leading to enhanced ethanol production. Mutant 647, selected as ethanol tolerant, was able to grow in medium containing 4% ethanol. During the early stage of the exponential growth phase, ethanol was produced as the main product, up to a concentration of about 9 g/liter. After 3 days of culture, 48.3 g (89% of the initial amount) of degraded cellulose per liter was fermented into 12.7 g of ethanol per liter. PMID- 16347825 TI - Enhanced Cellulose Fermentation by an Asporogenous and Ethanol-Tolerant Mutant of Clostridium thermocellum. AB - A mutant of Clostridium thermocellum isolated after UV mutagenesis and selection for resistance to fluoropyruvate was found to be asporogenous and ethanol tolerant. The mutant was also an ethanol hyperproducer, able to ferment 63 g of cellulose into 14.5 g of ethanol per liter of medium. The ratio of ethanol to total organic acids produced by the mutant was increased, and H(2) production was decreased. Culture conditions were optimized for ethanol production by the new strain. PMID- 16347826 TI - Inhibition of Yeast Growth by Octanoic and Decanoic Acids Produced during Ethanolic Fermentation. AB - The inhibition of growth by octanoic or decanoic acids, two subproducts of ethanolic fermentation, was evaluated in Saccharomyces cerevisiae and Kluyveromyces marxianus in association with ethanol, the main product of fermentation. In both strains, octanoic and decanoic acids, at concentrations up to 16 and 8 mg/liter, respectively, decreased the maximum specific growth rate and the biomass yield at 30 degrees C as an exponential function of the fatty acid concentration and increased the duration of growth latency. These toxic effects increased with a decrease in pH in the range of 5.4 to 3.0, indicating that the undissociated form is the toxic molecule. Decanoic acid was more toxic than octanoic acid. The concentrations of octanoic and decanoic acids were determined during the ethanolic fermentation (30 degrees C) of two laboratory media (mineral and complex) by S. cerevisiae and of Jerusalem artichoke juice by K. marxianus. Based on the concentrations detected (0.7 to 23 mg/liter) and the kinetics of growth inhibition, the presence of octanoic and decanoic acids cannot be ignored in the evaluation of the overall inhibition of ethanolic fermentation. PMID- 16347827 TI - Effect of fluoride on nitrification of a concentrated industrial waste. AB - The potential for biological nitrification of an industrial waste containing 4,000 mg of ammonia N (NH(4)-N) and 10,000 mg of fluoride per liter was investigated. Ammonium sulfate and sodium fluoride were tested in various combinations of 100 to 2,000 mg of NH(4)-N per liter and 0 to 5,000 mg of F per liter in suspended-growth stirred-tank reactors containing enriched cultures of nitrifying bacteria from a municipal sewage treatment plant. The stirred-tank reactors were fed once per day at a constant hydraulic retention period and cell retention time of 10 days. Temperature was 23 degrees C, and pH was 7.0 to 7.5. Clarified secondary effluent was used to make up feeds and to provide minor nutrients. Steady-state data, confirmed by mass balances, were obtained after five to six retention periods. In the absence of fluoride, nitrification efficiency was near 100% for up to 500 mg of NH(4)-N per liter. The influence of fluoride was studied at a low ammonia concentration (100 mg/liter) and exerted no significant effect on nitrification at concentrations of up to 200 mg/liter. Maximum effect of fluoride was reached at 800 mg of F per liter, and no greater inhibition was observed for up to 5,000 mg of F per liter. At the highest concentrations studied, ion pairing of ammonium and fluoride may exert a significant effect on kinetic coefficients. Kinetic analyses showed maximum specific substrate removal rates (q(max)) of NH(4)-N to be about 2.3 mg of N per mg of volatile suspended solids per day in the absence of fluoride and 0.85 mg of N per mg of volatile suspended solids per day in the presence of fluoride. The form of inhibition due to the presence of fluoride was shown to be not competitive, conforming to a mixed inhibition model. PMID- 16347828 TI - Chemokinetic Motility Responses of the Cyanobacterium Oscillatoria terebriformis. AB - Oscillatoria terebriformis, a gliding, filamentous, thermophilic cyanobacterium, exhibited an inhibition of gliding motility upon exposure to fructose. The observed response was transient, and the duration of nonmotility was directly proportional to the concentration of fructose. Upon resumption of motility, the rate of motility was also inversely proportional to the concentration of fructose. Sulfide caused a similar response. The effect of sulfide was specific and not due to either anoxia or negative redox potential. Exposure to glucose, acetate, lactate, or mat interstitial water did not elicit any motility response. PMID- 16347829 TI - Chemotaxis in Methanospirillum hungatei. AB - Methanospirillum hungatei gave a positive chemotactic response to acetate. PMID- 16347830 TI - Cytokinins in Azotobacter vinelandii Culture Medium. AB - Azotobacter vinelandii OP was grown to stationary phase in defined medium. The cell-free culture medium was analyzed for cytokinin content by XAD-2 and Sephadex LH-20 chromatography, thin-layer chromatography, tobacco callus bioassay, and enzyme immunoassay. Three cytokinin-active fractions were detected and tentatively identified as trans-zeatin, isopentenyladenosine, and isopentenyladenine. The total cytokinin activity was equivalent to 0.75 mug of kinetin per liter. PMID- 16347831 TI - Heat Shock Affects Permeability and Resistance of Bacillus stearothermophilus Spores. AB - [This corrects the article on p. 2517 in vol. 54.]. PMID- 16347833 TI - Acute copper and cupric ion toxicity in an estuarine microbial community. AB - Copper was acutely toxic to the estuarine microbial community of Middle Marshes, N.C. Under ambient water quality conditions, 10 mug of added total copper [Cu(II)] liter reduced the CFU bacterial abundance by up to 60% and inhibited the amino acid turnover rate (AATR) by as much as 30%. Copper toxicity, however, was a quantitative function of free cupric ion (Cu) activity that was not directly related to Cu(II) or ligand-bound copper. By using a nitrilotriacetic acid-cupric ion buffer to control pCu (-log Cu activity), it was found that an in situ pCu of 10.1 was bactericidal, reducing the CFU by 60%, but inhibited the AATR by only about 10%. A bacterial bioassay that was used to estimate the pCu in Cu(II) treated Middle Marshes samples indicated that less than 0.5% of added Cu(II) was in the free cupric ion form. CFU was a more sensitive indicator of low-level copper stress than was AATR. When tested at different times, native microbial community responses to acute cupric ion stress were quantitatively quite similar even when there were large differences in bacterial abundances and in situ metabolic rates. Variations were observed in response to Cu(II) treatments at different times, but these were likely due to differences in water quality, which would quantitatively influence the distribution of copper complexes that were present. Asymptotic response curves suggest that some degree of copper resistance exists in this community. At a pCu of 8, more than 2 orders of magnitude above the minimum inhibitory level, the CFU was still 5 to 10% and the AATR was about 3% of the control values. PMID- 16347832 TI - Effect of emulsan on biodegradation of crude oil by pure and mixed bacterial cultures. AB - Crude oil was treated with purified emulsan, the heteropolysaccharide bioemulsifier produced by Acinetobacter calcoaceticus RAG-1. A mixed bacterial population as well as nine different pure cultures isolated from various sources was tested for biodegradation of emulsan-treated and untreated crude oil. Biodegradation was measured both quantitatively and qualitatively. Recovery of CO(2) from mineralized C-labeled substrates yielded quantitative data on degradation of specific compounds, and capillary gas chromatography of residual unlabeled oil yielded qualitative data on a broad spectrum of crude oil components. Biodegradation of linear alkanes and other saturated hydrocarbons, both by pure cultures and by the mixed population, was reduced some 50 to 90% after emulsan pretreatment. In addition, degradation of aromatic compounds by the mixed population was reduced some 90% in emulsan-treated oil. In sharp contrast, aromatic biodegradation by pure cultures was either unaffected or slightly stimulated by emulsification of the oil. PMID- 16347834 TI - Effects of prechilling and sequential washing on enumeration of microorganisms from refuse. AB - Techniques were evaluated for formation of a liquid inoculum from shredded municipal refuse, including chilling the refuse at 4 degrees C prior to blending and multiple washing and blending cycles. The average count of cellulolytic bacteria from six different detachment treatments was 5.1 x 10 cells per g (dry weight) of refuse with a range of 0.7 x 10 to 12.7 x 10 cells per g (dry weight). The liquid obtained from blending the refuse in phosphate buffer followed by hand squeezing was the selected detachment procedure. The inoculum formation procedure was validated by the addition of ruminal cellulolytic bacteria to refuse and recovery of the cellulolytic bacteria by most-probable-number enumerations. The ratio of measured to expected cell counts among tests in which different volumes of ruminal fluid were added to refuse ranged from 2.7 to 14.4. There was no evidence of anaerobic cellulolytic fungi in a refuse sample. PMID- 16347835 TI - Bacterial population development and chemical characteristics of refuse decomposition in a simulated sanitary landfill. AB - Population development of key groups of bacteria involved in municipal refuse conversion to methane was measured from the time of initial incubation through the onset of methane production. Hemicellulolytic bacteria, cellulolytic bacteria, hydrogen-producing acetogens, and acetate- and H(2)-plus-CO(2) utilizing methanogens were enumerated by the most-probable-number technique with media containing oat spelt xylan, ball-milled cellulose, butyrate, acetate, and H(2) plus CO(2), respectively. Refuse decomposition was monitored in multiple replicate laboratory-scale sanitary landfills. A laboratory-scale landfill was dismantled weekly for microbial and chemical analysis. Leachate was neutralized and recycled to ensure methanogenesis. The methane concentration of the sampled containers increased to 64% by day 69, at which time the maximum methane production rate, 929 liters of CH(4) per dry kg-year, was measured. Population increases of 2, 4, 5, 5, and 6 orders of magnitude were measured between fresh refuse and the methane production phase for the hemicellulolytic bacteria, cellulolytic bacteria, butyrate-catabolizing acetogens, and acetate- and H(2) CO(2)-utilizing methanogens, respectively. The cellulolytic bacteria and acetogens increased more slowly than the methanogens and only after the onset of methane production. The initial decrease in the pH of the refuse ecosystem from 7.5 to 5.7 was attributed to the accumulation of acidic end products of sugar fermentation, to the low acid-consuming activity of the acetogenic and methanogenic bacteria, and to levels of oxygen and nitrate in the fresh refuse sufficient for oxidation of only 8% of the sugars to carbon dioxide and water. Cellulose and hemicellulose decomposition was most rapid after establishment of the methanogenic and acetogenic populations and a reduction in the initial accumulation of carboxylic acids. A total of 72% of these carbohydrates were degraded in the container sampled after 111 days. Initially acetate utilization, but ultimately polymer hydrolysis, limited the rate of refuse conversion to methane. Microbial and chemical composition data were combined to formulate an updated description of refuse decomposition in four phases: an aerobic phase, an anaerobic acid phase, an accelerated methane production phase, and a decelerated methane production phase. PMID- 16347836 TI - Transformation of Metalaxyl by the Fungus Syncephalastrum racemosum. AB - The fungus Syncephalastrum racemosum (Cohn) Schroeter was found to transform the fungicide metalaxyl [N-(2,6-dimethylphenyl)-N-(methoxyacetyl)-alanine methyl ester] in pure culture. After 21 days of incubation in a basal medium amended with 5 mug of metalaxyl per ml, more than 80% of the compound was transformed by the fungus. The transformation rates decreased as the concentrations of metalaxyl increased from 5 to 100 mug/ml. No transformation was observed when the concentration of metalaxyl was higher than 200 mug/ml. Two isomeric metabolites and a mixture of two other isomeric metabolites were isolated from the organic extract of the growth medium and identified as N-(2-methyl-6-hydroxymethylphenyl) N- and N-(2-hydroxymethyl-6-methylphenyl)-N-(methoxyacetyl)-alanine methyl ester and N-(3-hydroxy- and N-(5-hydroxy-2,6-dimethyl-phenyl)-N-(methoxyacetyl)-alanine methyl ester according to their mass-spectral and nuclear magnetic resonance spectral characteristics. Benzylic hydroxylation of the methyl side chains and/or aromatic hydroxylation appeared to be the major reactions involved in the metabolism of metalaxyl. PMID- 16347837 TI - Photodynamic production of superoxide in vitro by altertoxins in the presence of reducing agents. AB - Superoxide production by the three 4,9-dihydroxyperylene-3,10-quinone fungal toxins, altertoxins I, II, and III, was stimulated on illumination with broad spectrum light. As determined previously for cercosporin, superoxide production by illuminated altertoxins was increased by the addition of the reducing substances ergothioneine or urate; ascorbate also effectively increased superoxide production. Illuminated urate alone engendered some superoxide production. PMID- 16347838 TI - Stochastic models of soil denitrification. AB - Soil denitrification is a highly variable process that appears to be lognormally distributed. This variability is manifested by large sample coefficients of variation for replicate estimates of soil core denitrification rates. Deterministic models for soil denitrification have been proposed in the past, but none of these models predicts the approximate lognormality exhibited by natural denitrification rate estimates. In this study, probabilistic (stochastic) models were developed to understand how positively skewed distributions for field denitrification rate estimates result from the combined influences of variables known to affect denitrification. Three stochastic models were developed to describe the distribution of measured soil core denitrification rates. The driving variables used for all the models were denitrification enzyme activity and CO(2) production rates. The three models were distinguished by the functional relationships combining these driving variables. The functional relationships used were (i) a second-order model (model 1), (ii) a second-order model with a threshold (model 2), and (iii) a second-order saturation model (model 3). The parameters of the models were estimated by using 12 separate data sets (24 replicates per set), and their abilities to predict denitrification rate distributions were evaluated by using three additional independent data sets of 180 replicates each. Model 2 was the best because it produced distributions of denitrification rate which were not significantly different (P > 0.1) from distributions of measured denitrification rates. The generality of this model is unknown, but it accurately predicted the mean denitrification rates and accounted for the stochastic nature of this variable at the site studied. The approach used in this study may be applicable to other areas of ecological research in which accounting for the high spatial variability of microbiological processes is of interest. PMID- 16347839 TI - Field study comparing growth and viability of a population of phototrophic bacteria. AB - The growth and viability of an anoxygenic, phototrophic bacterial community in the hypolimnion of Zaca Lake, Calif., were compared throughout the summer. The community is dominated by a single species, "Thiopedia rosea," that inhabits the entire hypolimnion (6 to 8 m) for approximately 11 months. Suboptimal conditions in the hypolimnion (extremely low light intensity, high or low H(2)S levels) result in zero or extremely low growth rates (doubling times > 1 month) for most of the population, most of the time, yet cells remain viable and capable of high specific growth rates (doubling times of 1 to 10 days) when placed under favorable conditions (higher light intensities and temperatures). We first conclude that phototrophic bacterial populations in situ may frequently exist in a viable yet nongrowing state. Second, the viability of cells is likely to be reduced with depth owing to higher concentrations of potentially toxic chemicals and to changes in the physiological state associated with the prolonged periods of darkness commonly found at the bottom of bacterial plates. PMID- 16347840 TI - Evidence from liposome encapsulation for transport-limited microbial metabolism of solid alkanes. AB - The recalcitrance of xenobiotics may be caused by an absence of transforming enzymes or by their inability to enter microbial cells. A nondestructive method for differentiating between these two possibilities is described. The solid n alkanes octadecane (C(18)) and hexatriacontane (C(36)) were encapsulated into phosphatidylcholine bilayers (liposomes). The uptake and metabolism rates of encapsulated and unencapsulated substrates were then compared. During 1 h at 25 degrees C, a Pseudomonas isolate took up 1.3% of radiolabeled and unencapsulated C(18) (solid state) versus 23.5% of labeled and encapsulated C(18). Growth at 25 degrees C occurred with an apparent k(s) of 2453 +/- 148 mg/liter. Liposome encapsulation decreased this K(s) to 60 +/- 12 mg/liter. At 34 degrees C, growth on C(18) (liquid state) occurred with an apparent K(s) of 819 +/- 83 mg/liter and on the readily available carbon source succinate, K(s) values were 80 +/- 10 and 13 +/- 7 mg/liter at 25 and 34 degrees C, respectively. At 25 degrees C, the isolate grew on C(36) with an apparent K(s) of 2,698 +/- 831 mg/liter. Liposome encapsulation decreased the K(s) more than 60-fold to 41 +/- 7 mg/liter, resulting in the complete utilization of 400 mg of C(36) per liter in 16 h. Since controls excluded the metabolic utilization of phosphatidylcholine, the results clearly identify transport limitation as the cause for C(36) recalcitrance. PMID- 16347841 TI - Pyrroloquinoline Quinone-Dependent Cytochrome Reduction in Polyvinyl Alcohol Degrading Pseudomonas sp. Strain VM15C. AB - A polyvinyl alcohol (PVA) oxidase-deficient mutant of Pseudomonas sp. strain VM15C, strain ND1, was shown to possess PVA dehydrogenase, in which pyrroloquinoline quinone (PQQ) functions as a coenzyme. The mutant grew on PVA and required PQQ for utilization of PVA as an essential growth factor. Incubation of the membrane fraction of the mutant with PVA caused cytochrome reduction of the fraction. Furthermore, it was found that in spite of the presence of PVA oxidase, the membrane fraction of strain VM15C grown on glucose without PQQ required PQQ for cytochrome reduction during incubation with PVA. The results provide evidence that PVA dehydrogenase couples with the electron transport chain of PVA-degrading bacteria but that PVA oxidase does not. PMID- 16347842 TI - DNA Probe for Identification of the Take-All Fungus, Gaeumannomyces graminis. AB - A 4.3-kilobase mitochondrial DNA fragment was cloned from Gaeumannomyces graminis var. tritici, the causative agent of take-all disease of wheat. Although this DNA fragment hybridized with all three varieties of G. graminis, it showed little homology with DNA from other fungi and thus should be useful for identification of Gaeumannomyces sp. recovered from infected plants. PMID- 16347843 TI - Isolation and Preliminary Characterization of Hydroxamic Acids Formed by Nitrogen Fixing Azotobacter chroococcum B-8. AB - The free-living diazotroph Azotobacter chroococcum B-8 responded to iron-limited growth conditions by forming hydroxamic acids and an 85,000-dalton outer membrane protein. The Fe(III)-binding hydroxamate compounds stimulated the growth of Arthrobacter flavescens JG-9 and gave a positive Csaky reaction for bound hydroxylamines. The hydroxamates were isolated from liquid cultures by benzyl alcohol extraction and purified by size exclusion chromatography and high performance liquid chromatography. Four high-performance liquid chromatography fractions, designated A, B, C, and D, had the characteristic hydroxamate absorption maximum at 420 to 423 nm, which did not shift over a pH range from 3.0 to 9.0. Cyclic voltammograms of the iron-hydroxamate complexes exhibited reduction potentials of -0.426 to -0.442 V for fractions A, B, and D and of -0.56 V for fraction C versus the normal hydrogen electrode at pH 8.0. Based on mass spectra, nominal molecular weights of 800 and 844 were assigned to ferrated compounds A and B, respectively. Reductive hydrolysis of compounds A and B in 57% hydriodic acid yielded ornithine as detected by gas chromatography-mass spectrometry. All of these physiological and chemical data strongly support the hypothesis that the high-affinity iron-binding compounds isolated from A. chroococcum B-8 are hydroxamic acids and probably function as siderophores for this diazotroph. PMID- 16347844 TI - Microbiological oxidation of ferrous iron at low temperatures. AB - Acidophilic iron-oxidizing bacteria were enriched from mine water samples with ferrous sulfate as the substrate at incubation temperatures in the range of 4 to 46 degrees C. After several subcultures at each test temperature except 46 degrees C, which was prohibitive to growth, the rates of iron oxidation were determined in batch cultures. The results yielded linear rates in a semilogarithmic scale. The rate constants of iron oxidation by growing cultures were fitted into the Arrhenius equation, which displayed linearity in the 4 to 28 degrees C range and yielded an activation energy value of 83 +/- 3 kJ/mol. PMID- 16347845 TI - Possible Involvement of Toluene-2,3-Dioxygenase in Defluorination of 3-Fluoro Substituted Benzenes by Toluene-Degrading Pseudomonas sp. Strain T-12. AB - Pseudomonas sp. strain T-12 cells in which the toluene-degradative pathway enzymes have been induced can transform many 3-fluoro-substituted benzenes to the corresponding 2,3-catechols with simultaneous elimination of the fluorine substituent as inorganic fluoride. Substrates for this transformation included 3 fluorotoluene, 3-fluorotrifluorotoluene, 3-fluorohalobenzenes, 3-fluoroanisole, and 3-fluorobenzonitrile. While 3-fluorotoluene and 3-fluoroanisole produced only defluorinated catechols, other substrates generated catechol products with and without the fluorine substituent. The steric size of the C-1 substituent affected the ratio of defluorinated to fluorinated catechols formed from a substrate. A mechanism for the defluorination reaction involving toluene-2,3-dioxygenase is proposed. PMID- 16347846 TI - Bioconversion of Gelatin to Methane by a Coculture of Clostridium collagenovorans and Methanosarcina barkeri. AB - A simple, stable, and transferable coculture of Clostridium collagenovorans and Methanosarcina barkeri that readily degraded gelatin into methane and carbon dioxide was developed. In monoculture, C. collagenovorans fermented all of the amino acids in gelatin except proline into acetate and carbon dioxide as the main products, with hydrogen, isovalerate, and isobutyrate detected in trace amounts (<1 mM). In coculture with M. barkeri, gelatin was transformed into methane and carbon dioxide, with varying levels of intermediary acetate formed as a function of incubation time. Various complex proteinaceous polymers could be readily transformed into methane and carbon dioxide at 30 to 40 degrees C by a stable coculture which did not require exogenous growth factor additions. In addition, the coculture was readily transferable and preserved in the viable state for long periods, and methanogenesis could be initiated rapidly without the need for exogenous pH control. PMID- 16347847 TI - Characterization of a Pseudomonas sp. Capable of Aniline Degradation in the Presence of Secondary Carbon Sources. AB - Pseudomonas strain K1 is a gram-negative rod which grows aerobically on minimal media containing aniline with a doubling time of 2 h at 30 degrees C. The half saturation parameter for aniline metabolism by aniline-grown cells was 3.8 mumol . liter. Concentrations of aniline as low as 50 nM were metabolized. Neither substituted anilines nor other aromatic compounds (other than aromatic amino acids) supported growth. Cells grew as fast on aniline as on nonaromatic substrates such as lactate. The aromatic ring was cleaved via the meta pathway. Catechol 2,3-oxygenase activity was induced by aniline, even in cultures containing alternative carbon sources such as lactate. Cultures grown on a mixture of aniline and lactate mineralized aniline in the presence of the second substrate. Lactate-grown cultures lacked catechol oxygenase activity, and resting cells from these cultures did not respire aniline. Resting cells from aniline grown cultures exhibited high respiratory activity upon the addition of aniline or catechol, some activity with toluidine, and no activity after addition of a wide variety of other aromatic compounds, including dihydroxybenzylamine, chloroanilines, ethylanilines, aminophenols, aminobenzoates, and dihydroxybenzoates. Although substituted anilines were not metabolized, 3-or 4 chloroaniline did induce the enzymes for aniline oxidation. PMID- 16347848 TI - Transformation by Complementation of an Adenine Auxotroph of the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium. AB - Swollen basidiospores of an adenine auxotroph of Phanerochaete chrysosporium were protoplasted with Novozyme 234 and transformed to prototrophy by using a plasmid containing the gene for an adenine biosynthetic enzyme from Schizophyllum commune. Transformation frequencies of 100 transformants per mug of DNA were obtained. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Analysis of conidia and basidiospores from transformants demonstrated that the transforming character was mitotically and meiotically stable on both selective and nonselective media. Genetic crosses between double mutants transformed for adenine prototrophy and other auxotrophic strains yielded Ade progeny, which indicated that integration occurred at a site(s) other than the resident adenine biosynthetic gene. PMID- 16347849 TI - Effects of ozone and sulfur dioxide on phyllosphere fungi from three tree species. AB - Short-term effects of ozone (O(3)) on phyllosphere fungi were studied by examining fungal populations from leaves of giant sequoia (Sequoiadendron giganteum (Lindl.) Buchholz) and California black oak (Quercus kelloggii Newb.). Chronic effects of both O(3) and sulfur dioxide (SO(2)) were studied by isolating fungi from leaves of mature Valencia orange (Citrus sinensis L.) trees. In this chronic-exposure experiment, mature orange trees were fumigated in open-top chambers at the University of California, Riverside, for 4 years with filtered air, ambient air plus filtered air (1:1), ambient air, or filtered air plus SO(2) at 9.3 parts per hundred million. Populations of Alternaria alternata (Fr.) Keissler and Cladosporium cladosporioides (Fres.) de Vries, two of the four most common fungi isolated from orange leaves, were significantly reduced by chronic exposure to ambient air. In the short-term experiments, seedlings of giant sequoia or California black oak were fumigated in open-top chambers in Sequoia National Park for 9 to 11 weeks with filtered air, ambient air, or ambient air plus O(3). These short-term fumigations did not significantly affect the numbers of phyllosphere fungi. Exposure of Valencia orange trees to SO(2) at 9.3 parts per hundred million for 4 years reduced the number of phyllosphere fungi isolated by 75% compared with the number from the filtered-air treatment and reduced the Simpson diversity index value from 3.3 to 2.5. A significant chamber effect was evident since leaves of giant sequoia and California black oak located outside of chambers had more phyllosphere fungi than did seedlings within chambers. Results suggest that chronic exposure to ambient ozone or SO(2) in polluted areas can affect phyllosphere fungal communities, while short-term exposures may not significantly disturb phyllosphere fungi. PMID- 16347850 TI - Effect of light on the cell cycle of a marine synechococcus strain. AB - Light-dependent regulation of cell cycle progression in the marine cyanobacterium Synechococcus strain WH-8101 was demonstrated through the use of flow cytometry. Our results show that, similar to eucaryotic cells, marine Synechococcus spp. display two gaps in DNA synthesis, at the beginning and at the end of the cell cycle. Progression through each of these gaps requires light, and their durations lengthen under light limitation. PMID- 16347851 TI - Survey of the anaerobic biodegradation potential of organic chemicals in digesting sludge. AB - The degradation potential of 77 organic chemicals under methanogenic conditions was examined with an anaerobic digesting sludge from the United Kingdom. Degradation was assessed in terms of net total gas (CH(4) plus CO(2)) produced, expressed as a percentage of the theoretical production (ThGP). The compounds tested were selected from various chemical groups and included substituted phenols and benzoates, pesticides, phthalic acid esters, homocyclic and heterocyclic ring compounds, glycols, and monosubstituted benzenes. The results obtained were in good agreement with published surveys of biodegradability in U.S. digesting sludges and other methanogenic environments. In general, the presence of chloro or nitro groups inhibited anaerobic gas production, while carboxyl and hydroxyl groups facilitated biodegradation. The relationship between substituent position and susceptibility to methanogenic degradation was compound dependent. The following chemicals were completely degraded (>/=80% ThGP) at a concentration of 50 mg of carbon per liter: phenol, 2-aminophenol, 4-cresol, catechol, sodium benzoate, 4-aminobenzoic acid, 3-chlorobenzoic acid, phthalic acid, ethylene glycol, diethylene glycol, triethylene glycol, sodium stearate, and quinoline. 3-Cresol, 4-chlorobenzoic acid, dimethyl phthalate, and pyridine were partially degraded. Although the remaining chemicals tested were either persistent or toxic, their behavior may differ at more environmentally realistic chemical-to-biomass ratios. Our findings suggest that biodegradability assessments made with sludge from one source can be extrapolated to sludge from another source with a reasonable degree of confidence and should help in predicting the fate of an organic chemical during the anaerobic digestion of sewage sludge. PMID- 16347852 TI - Methanogenesis from ethanol by defined mixed continuous cultures. AB - Methanogenesis from ethanol by defined mixed continuous cultures was studied. Under sulfate-free conditions, a Desulfovibrio strain was used as the ethanol degrading species producing acetic acid and hydrogen. In a two-membered mutualistic coculture, the hydrogen was converted to methane by a Methanobacterium sp. and pH was maintained at neutrality by the addition of alkali. Introduction of a third species, the acetate-utilizing Methanosarcina mazei, obviated the need for external pH control. Methanogenesis by the co-and triculture was studied at various dilution rates in the steady state. The mutualistic coculture performed like a composite single species, as predicted from the theory of mutualistic interactions. Coupling between the mutualistic coculture and the acetate-utilizing methanogen was less tight. Increasing the dilution rate destabilized the triculture; at low dilution rates, instability was soon recovered, but at higher dilution rates imbalance between the rates of production and removal of acetic acid led to a drop in pH. Flocs formed in the triculture. An annulus of the Methanobacterium sp. and Desulfovibrio sp. was retained around the Methanosarcina sp. by strands of material probably derived from the Methanosarcina sp. PMID- 16347853 TI - Survival of Azorhizobium caulinodans in the Soil and Rhizosphere of Wetland Rice under Sesbania rostrata-Rice Rotation. AB - The survival of indigenous and introduced strains of Azorhizobium caulinodans in flooded soil and in the rice rhizosphere, where in situ Sesbania rostrata was incorporated before the rice crop, is reported. The azorhizobia studied were both root and stem nodulating. In a pot experiment, two crop cycles each of inoculated and noninoculated Sesbania-rice were compared with two crop cycles of flooded fallow-rice. In a field experiment, the effect of repeated incorporation of in situ S. rostrata in the Sesbania-rice sequence was studied. Soils in which inoculated S. rostrata was incorporated contained about 3,000 times more azorhizobia than did soils in the flooded fallow treatment and about 50 times more azorhizobia than did soils in the noninoculated Sesbania treatment. Azorhizobial numbers in the inoculated Sesbania treatment declined toward rice harvest but remained much higher than in the flooded fallow-rice treatment. Repeated incorporation of S. rostrata increased the population density of indigenous soil azorhizobia, whereas the population of inoculated strain ORS571 (Str Spc) declined to an undetectable level; this finding suggested low competitiveness by the introduced strain. In the incorporated Sesbania treatment, the rice rhizosphere harbored significantly more A. caulinodans and supported higher nitrogenase activity per plant than did the rhizosphere of the flooded fallow-rice treatment. Sterile rice seedlings inoculated with A. caulinodans showed nitrogenase activity comparable to that of seedlings inoculated with Azospirillum lipoferum 34H, a rice root isolate. Rhizobia from Sesbania aculeata, Sesbania sesban, a Trifolium sp., and Vigna unguiculata did not support appreciable nitrogenase activity. PMID- 16347854 TI - Degradation of 4-Chlorobenzoate by Facultatively Alkalophilic Arthrobacter sp. Strain SB8. AB - A facultative alkalophile capable of utilizing 4-chlorobenzoate (4-CBA), strain SB8, was isolated from soil with an alkaline medium (pH 10.0) containing the haloaromatic compound as the carbon source. The strain, identified as an Arthrobacter sp., showed rather extensive 4-CBA-degrading ability. 4-CBA utilization by the strain was possible in the alkaline medium containing up to 10 g of the compound per liter. The 4-CBA-dechlorinating activity of resting cells was almost completely uninhibited by substrate concentrations up to 150 mM. The bacterium dehalogenated 4-CBA in the initial stage of the degradation and metabolized the compound via 4-hydroxybenzoate and protocatechuate. O(2) was needed for 4-CBA dechlorination by resting cells but not by cell extracts. O(2) was inhibitory to the 4-CBA dechlorination activity of cell extracts. These facts suggest dechlorination of 4-CBA by halide hydrolysis and an energy requirement for the transport of 4-CBA into cells. PMID- 16347855 TI - Coupling of Solar Energy to Hydrogen Peroxide Production in the Cyanobacterium Anacystis nidulans. AB - Hydrogen peroxide production by blue-green algae (cyanobacteria) under photoautotrophic conditions is of great interest as a model system for the bioconversion of solar energy. Our experimental system was based on the photosynthetic reduction of molecular oxygen with electrons from water by Anacystis nidulans 1402-1 as the biophotocatalyst and methyl viologen as a redox intermediate. It has been demonstrated that the metabolic conditions of the algae in their different growth stages strongly influence the capacity for hydrogen peroxide photoproduction, and so the initial formation rate and net peroxide yield became maximum in the mid-log phase of growth. The overall process can be optimized in the presence of certain metabolic inhibitors such as iodoacetamide and p-hydroxymercuribenzoate, as well as by permeabilization of the cellular membrane after drastic temperature changes and by immobilization of the cells in inert supports such as agar and alginate. PMID- 16347856 TI - Kinetics of Acetate Utilization by Two Thermophilic Acetotrophic Methanogens: Methanosarcina sp. Strain CALS-1 and Methanothrix sp. Strain CALS-1. AB - The kinetics of acetate utilization were examined for washed concentrated cell suspensions of two thermophilic acetotrophic methanogens isolated from a 58 degrees C anaerobic digestor. Progress curves for acetate utilization by cells of Methanosarcina sp. strain CALS-1 showed that the utilization rate was concentration independent (zero order) above concentrations near 3 mM and that acetate utilization ceased when a threshold concentration near 1 mM was reached. Acetate utilization by cells of Methanothrix sp. strain CALS-1 was concentration independent down to 0.1 to 0.2 mM, and threshold values of 12 to 21 muM were observed. Typical utilization rates in the concentration-independent stage were 210 and 130 nmol min mg of protein for the methanosarcina and the methanothrix, respectively. These results are in agreement with a general model in which high acetate concentrations favor Methanosarcina spp., while low concentrations favor Methanothrix spp. However, acetate utilization by these two strains did not follow simple Michaelis-Menton kinetics. PMID- 16347857 TI - 3-nitrobenzenesulfonate, 3-aminobenzenesulfonate, and 4-aminobenzenesulfonate as sole carbon sources for bacteria. AB - Aerobic, carbon-limited, enrichment cultures containing 3-aminobenzenesulfonate or 3-nitrobenzenesulfonate as the sole source of carbon and energy yielded growth and complete substrate disappearance. Pure cultures of putative pseudomonads were isolated which utilized these compounds quantitatively. Degradation was compared with that of 2- and 4-aminobenzenesulfonate. PMID- 16347858 TI - Threshold acetate concentrations for acetate catabolism by aceticlastic methanogenic bacteria. AB - Marked differences were found for minimum threshold concentrations of acetate catabolism by Methanosarcina barkeri 227 (1.180 mM), Methanosarcina mazei S-6 (0.396 mM), and a Methanothrix sp. (0.069 mM). This indicates that the aceticlastic methanogens responsible for the conversion of acetate to methane in various ecosystems might be different, depending on the prevailing in situ acetate concentrations. PMID- 16347859 TI - Hydroxylation and Dechlorination of Tetrachlorohydroquinone by Rhodococcus sp. Strain CP-2 Cell Extracts. AB - A cell extract of a polychlorophenol-degrading bacterium, Rhodococcus sp. strain CP-2, isolated from chlorophenol-contaminated soil, was shown to dechlorinate tetrachlorohydroquinone, the first intermediate in pentachlorophenol and 2,3,5,6 tetrachlorophenol degradation. Degradation of tetrachlorohydroquinone was catalyzed by a soluble enzyme(s). The reaction sequence for complete dechlorination involved hydroxylation and three reductive dechlorinations, producing 1,2,4-trihydroxybenzene. All chlorines were thus removed from the polychlorinated compound before ring cleavage. PMID- 16347860 TI - Rhizobium strain identification and quantification in commercial inoculants by immunoblot analysis. AB - An immunoblot procedure for the strain-specific quantitative analysis of commercial Rhizobium inoculants was developed. The technique greatly reduced the time required for inoculant analysis. Correlation between immunoblot analysis and traditional plant nodule grow-out most-probable-number techniques was r = 0.90 for 16 commercial alfalfa inoculants tested. PMID- 16347861 TI - In Vitro Evolution and Preliminary Characterization of a Cadmium-Resistant Population of Chlamydomonas reinhardtii. AB - A cadmium-tolerant population of Chlamydomonas reinhardtii was derived from a Cd sensitive cell wall-deficient strain by long-term selection in liquid culture. A comparison of Cd-sensitive and Cd-tolerant cells revealed that Cd tolerance was due to genetically determined alterations of metabolism rather than to increased efficiency of a detoxification system. PMID- 16347862 TI - Scanning Electron Microscopy of Invasion of Apple Leaves and Blossoms by Pseudomonas syringae pv. syringae. AB - Scanning electron microscopy indicated that Pseudomonas syringae pv. syringae L795 entered leaves through stomata and multiplied in the substomatal chambers. Strain L195 applied to blossoms colonized stigmas and also occurred in intercellular spaces of styles. Nonpathogenic strain L796 failed to colonize blossoms. This study suggests that inoculum of pathogenic P. syringae pv. syringae builds up on apple leaves and blossoms. PMID- 16347863 TI - Dynamics of Microbial Populations during Fermentation of Wines from the Utiel Requena Region of Spain. AB - The dynamics of fungi, yeasts, and lactic acid bacteria during fermentation of four musts were studied. Fungi disappeared quickly in the fermenting must. The lactic acid bacteria population diminished during alcoholic fermentation, then they increased and performed malolactic fermentation. Yeasts grew quickly, reaching maximum populations at different times depending on the vinification treatment. PMID- 16347864 TI - Ruminal ciliated protozoa in bison. AB - [This corrects the article on p. 2734 in vol. 54.]. PMID- 16347865 TI - Enhanced Conversion of Lactose to Glycerol by Kluyveromyces fragilis Utilizing Whey Permeate as a Substrate. AB - Kluyveromyces fragilis (CBS 397) is a nonhalophilic yeast which is capable of lactose utilization from whey permeate and high glycerol production under anaerobic growth conditions. However, the optimum yields of glycerol (11.6 mg/ml of whey permeate medium) obtained in this study occurred only in the presence of 1% Na(2)SO(3) as a steering agent. The use of other concentrations of Na(2)SO(3), as well as 5% NaCl and 1% ascorbic acid, had no or detrimental effects on cell growth, lactose utilization, and glycerol production. Glycerol yields were greater in cultures grown from a light inoculum of K. fragilis than in cultures in which a resuspended mass of cells was introduced into the medium. The results of this study suggest that this strain of K. fragilis may be useful commercially in the utilization of cheese whey lactose and the concomitant production of glycerol. PMID- 16347866 TI - Characterization of exopolysaccharides produced by plant-associated fluorescent pseudomonads. AB - A total of 214 strains of plant-associated fluorescent pseudomonads were screened for the ability to produce the acidic exopolysaccharide (EPS) alginate on various solid media. The fluorescent pseudomonads studied were saprophytic, saprophytic with known biocontrol potential, or plant pathogenic. Approximately 10% of these strains exhibited mucoid growth under the conditions used. The EPSs produced by 20 strains were isolated, purified, and characterized. Of the 20 strains examined, 6 produced acetylated alginate as an acidic EPS. These strains included a Pseudomonas aeruginosa strain reported to cause a dry rot of onion, a strain of P. viridiflava with soft-rotting ability, and four strains of P. fluorescens. However, 12 strains of P. fluorescens produced a novel acidic EPS (marginalan) composed of glucose and galactose (1:1 molar ratio) substituted with pyruvate and succinate. Three of these strains were soft-rotting agents. Two additional soft rotting strains of P. fluorescens produced a third acidic novel EPS composed of rhamnose, mannose, and glucose (1:1:1 molar ratio) substituted with pyruvate and acetate. When sucrose was present as the primary carbon source, certain strains produced the neutral polymer levan (a fructan) rather than an acidic EPS. Levan was produced by most strains capable of synthesizing alginate or the novel acidic EPS containing rhamnose, mannose, and glucose but not by strains capable of marginalan production. It is now evident that the group of bacteria belonging to the fluorescent pseudomonads is capable of elaborating a diverse array of acidic EPSs rather than solely alginate. PMID- 16347867 TI - Production of Poly-beta-Hydroxyalkanoic Acid by Pseudomonas cepacia. AB - The possibility of using the nutritionally versatile bacterium Pseudomonas cepacia to produce poly-beta-hydroxyalkanoic acid was evaluated. Chemostat culture showed that growth of P. cepacia became nitrogen limited when the molar carbon-to-nitrogen ratio of the medium fed into the fermentor was above 15. When grown under nitrogen limitation in batch culture with fructose as the sole source of carbon, P. cepacia accumulated poly-beta-hydroxybutyric acid (PHB) in excess of 50% of the dry weight of its biomass. In batch culture, almost no PHB was produced until the onset of nitrogen limitation. After this point, PHB was produced at a linear rate of 0.12 g liter h (from a constant value of 1.6 g of cellular protein liter). PHB produced by P. cepacia had a weight-average molecular weight of 5.37 x 10 g mol and a polydispersivity index of 3.9. Poly(beta-hydroxybutyric acid-beta-hydroxyvaleric acid) copolymer was produced with a poly-beta-hydroxybutyric acid-poly-beta-hydroxyvaleric acid ratio of up to 30% by weight when propionic acid was added to the medium. PMID- 16347868 TI - Influence of alternate electron acceptors on the metabolic fate of hydroxybenzoate isomers in anoxic aquifer slurries. AB - The biodegradation of hydroxybenzoate isomers was investigated with samples obtained from two sites within a shallow anoxic aquifer. The metabolic fates of the substrates were compared in denitrifying, sulfate-reducing, and methanogenic incubations. Under the latter two conditions, phenol was detected as a major intermediate of p-hydroxybenzoate, but no metabolites were initially found with m or o-hydroxybenzoate. However, benzoate accumulation was noted when metabolic inhibitors were used with these samples. About 9 to 17 days was required for >95% removal of the parent isomers under these conditions. When aquifer slurries were amended with nitrate, the equivalent removal of the hydroxybenzoates occurred within 4 days. In the denitrifying incubations, phenol was formed from all three hydroxybenzoates and accounted for about 30% of the initial substrate amendment. No benzoate was measured in these samples. All metabolites were identified by chromatographic mobility, mass spectral profiles, or both. Autoclaved controls were uniformly incapable of transforming the parent substrates. These results suggest that the anaerobic fate of hydroxybenzoate isomers depends on the relative substitution pattern and the prevailing ecological conditions. Furthermore, since these compounds are central metabolites formed during the breakdown of many aromatic chemicals, our findings may help provide guidelines for the reliable extrapolation of metabolic fate information from diverse anaerobic environments. PMID- 16347869 TI - Physical degradation of lignified stem tissues by ruminal fungi. AB - Ruminal bacteria or fungi were selected by the addition of cycloheximide or streptomycin and penicillin, respectively, to ruminal fluid, and the weakening and degradation of lignified tissues in alfalfa and Bermuda grass stems by these treatments and whole ruminal fluid were evaluated in vitro. Dry weight loss in alfalfa was similar for whole ruminal fluid and streptomycin-penicillin treatment, whereas that with streptomycin-penicillin treatment was significantly higher (P 0.4-mum direct count regardless of media, season of sampling, or soil depth. In the same soil samples, the viable nodulating populations of biovar trifolii determined by the plant infection soil dilution technique ranged between 1 and 10% of the >0.4-mum direct-immunofluorescence count of biovar trifolii. The <0.4 mum cell populations of both total soil bacteria and biovar trifolii changed abruptly between the 10- to 15-cm and 15- to 20-cm soil depth increments, increasing from 5 to 20% and from 20 to 50%, respectively, of their direct-count totals. The increase in density of the small-cell population corresponded to a significant increase in soil bulk density (1.07 to 1.21 g cm). The percent contribution of the <0.4-mum direct count to individual serogroup totals increased with soil depth by approximately 2-fold (39 to 87%) for serogroups 17 and 21 and by 12-fold (6 to 75%) for serogroups 6 and 36. PMID- 16347897 TI - Betaine fermentation and oxidation by marine desulfuromonas strains. AB - Two bacterial strains were dominant in anaerobic enrichment cultures with betaine (N,N,N-trimethylglycine) as a substrate and intertidal mud as an inoculum. One was a coccoid bacterium which was a trimethylamine (TMA)-fermenting methanogen similar to Methanococcoides methylutens. The other strain, a rod-shaped, gram negative, motile bacterium, fermented betaine. On the basis of its ability to oxidize acetate and ethanol to CO(2) with sulfur as an electron acceptor, its inability to reduce sulfate and sulfite, its morphology, the presence of c-type cytochromes, and other characteristics, the isolated strain PM1 was identified as Desulfuromonas acetoxidans. Although only malate and fumarate were known as substrates for fermentative growth of this species, the type strain (DSM 684) also fermented betaine. Strain PM1 grew with a doubling time of 9.5 h at 30 degrees C on betaine and produced approximately 1 mol of TMA per mol of betaine, 0.75 mol of acetate, and presumably CO(2) as fermentation products but only in the presence of selenite (100 nM). In this fermentation, betaine is probably reductively cleaved to TMA and acetate, and part of the acetate is then oxidized to CO(2) to provide the reducing equivalents for the initial cleavage reaction. In the presence of sulfur, betaine was converted to TMA and presumably CO(2) with the formation of sulfide; then, only traces of acetate were produced. PMID- 16347898 TI - Isolation and Characterization of Mutants of Clostridium acetobutylicum ATCC 824 Deficient in Acetoacetyl-Coenzyme A:Acetate/Butyrate:Coenzyme A-Transferase (EC 2.8.3.9) and in Other Solvent Pathway Enzymes. AB - Mutants of Clostridium acetobutylicum ATCC 824 exhibiting resistance to 2 bromobutyrate or rifampin were isolated after nitrosoguanidine treatment. Mutants were screened for solvent production by using an automated alcohol test system. Isolates were analyzed for levels of butanol, ethanol, acetone, butyrate, acetate, and acetoin in stationary-phase batch cultures. The specific activities of NADH- and NADPH-dependent butanol dehydrogenase and butyraldehyde dehydrogenase as well as those of acetoacetyl-coenzyme A:acetate/butyrate:coenzyme A-transferase (butyrate-acetoacetate coenzyme A transferase [EC 2.8.3.9]) (CoA-transferase), butyrate kinase, and phosphotransbutyrylase were measured at the onset of stationary phase. Rifampin resistant strain D10 and 2-bromobutyrate mutant R were found to be deficient in only CoA-transferase, while several other mutants exhibited reduced butyraldehyde dehydrogenase and butanol dehydrogenase activities as well. The colony morphology of 2-bromobutyrate mutant R was similar to that of the parent on RCM medium; however, it had about 1/10 the level of CoA-transferase and increased levels of butanol dehydrogenase and butyraldehyde dehydrogenase. A nonsporulating, spontaneously derived degenerated strain exhibited reduced levels of butyraldehyde dehydrogenase, butanol, dehydrogenase, and CoA-transferase compared with those of the original strain. When C. acetobutylicum ATCC 824 was grown on medium containing low levels of 2-bromobutyrate, an altered colony morphology was observed. Not all strains resistant to 2-bromobutyrate (12 mM) were non-solvent producing strains. PMID- 16347899 TI - Effects of Organic Acid Anions on the Growth and Metabolism of Syntrophomonas wolfei in Pure Culture and in Defined Consortia. AB - The effects of organic acid anions on the growth of Syntrophomonas wolfei was determined by varying the initial concentration of the acid anion in the medium. The addition of 15 mM acetate decreased the growth rate of a butyrate catabolizing coculture containing Methanospirillum hungatei from 0.0085 to 0.0029 per hour. Higher initial acetate concentrations decreased the butyrate degradation rate and the yield of cells of S. wolfei per butyrate degraded. Inhibition was not due to the counter ion or the effect of acetate on the methanogen. Initial acetate concentrations above 25 mM inhibited crotonate-using pure cultures and cocultures of S. wolfei. Benzoate and lactate inhibited the growth of S. wolfei on crotonate in pure culture and coculture. Lactate was an effective inhibitor of S. wolfei cultures at concentrations greater than 10 mM. High concentrations of acetate and lactate altered the electron flow in crotonate catabolizing cocultures, resulting in the formation of less methane and more butyrate and caproate. The inclusion of the acetate-using methanogen, Methanosarcina barkeri, in a methanogenic butyrate-catabolizing coculture increased both the yield of S. wolfei cells per butyrate degraded and the efficacy of butyrate degradation. Butyrate degradation by acetate-inhibited cocultures occurred only after the addition of Methanosarcina barkeri. These results showed that the metabolism of S. wolfei was inhibited by high levels of organic acid anions. The activity of acetate-using methanogens is important for the syntrophic degradation of fatty acids when high levels of acetate are present. PMID- 16347900 TI - Description of an estuarine methylotrophic methanogen which grows on dimethyl sulfide. AB - Characteristics of an obligately methylotrophic coccoid methanogen (strain GS-16) previously isolated from estuarine sediment are described. Growth was demonstrated on dimethyl sulfide (DMS) or trimethylamine (TMA), but not on methane thiol, methane thiol plus hydrogen, dimethyl disulfide, or methionine. DMS-grown cells were able to metabolize DMS and TMA simultaneously when inoculated into media containing substrate levels of these compounds. However, TMA-grown cells could not metabolize [C]DMS to CH(4), although they could convert [C]methanol to CH(4). These results suggest that metabolism of DMS proceeds along a somewhat different route than that of TMA and perhaps also that of methanol. The organism exhibited doubling times of 23 and 32 h for growth (25 degrees C) in mineral media on TMA and DMS, respectively. Doubling times were more rapid ( approximately 6 h) when the organisms were grown on TMA in complex broth. In mineral media, the fastest growth on DMS occurred between pH levels of 7.0 and 8.7, at 29 degrees C, and with 0.2 to 0.4 M Na and 0.04 M Mg. Somewhat different results occurred for growth on TMA in complex broth. Cells had a moles percent G+C value of 44.5% for their DNA. Growth on DMS, TMA, and methanol yielded stable carbon isotope fractionation factors of 1.044, 1.037, and 1.063, respectively. Fractionation factors for hydrogen were 1.203 (DMS) and 1.183 (TMA). PMID- 16347901 TI - Comparative Replication of Lymantria dispar Nuclear Polyhedrosis Virus Strains in Three Continuous-Culture Cell Lines. AB - We compared the replication of the gypsy moth (Lymantria dispar) nuclear polyhedrosis virus in two new cell lines, from embryos and fat body of L. dispar, and in a previously available ovarian cell line. Three virus isolates (the Hamden strain [LDP-67] used commercially as GYPCHEK, a plaque-purified clone of Hamden [5-7d], and an isolate from Abington, Mass. [Ab]) were each tested on the three cell lines. The fat-body-derived cell line proved best in terms of occlusion body production for all three virus strains, with the highest yield produced by the Abington strain. On the basis of these results, we conclude that a more efficient in vitro production of gypsy moth virus can be obtained by using the fat body cell line in conjunction with the Abington strain of the virus. PMID- 16347902 TI - Vertical and horizontal variations in the physiological diversity of the aerobic chemoheterotrophic bacterial microflora in deep southeast coastal plain subsurface sediments. AB - Aerobic chemoheterotrophic bacteria were isolated from surface soils and coastal plain subsurface (including deep aquifer) sediments (depths to 265 m) at a study site near Aiken, S.C., by plating on concentrated and dilute media. Morphologically distinct colonies were purified, and their responses to 21 selected physiological tests were determined. These isolates were quite diverse; 626 physiologically distinct types (i.e., types with a unique pattern of responses to the 21 tests) were detected among the 1,112 isolates obtained. Physiologically distinct types were isolated on concentrated and dilute media (only 11% overlap between the groups); isolates from surface soils and subsurface sediments were also quite different (only 3% overlap). The surface soil isolates more readily utilized all but 1 of 12 carbon sources offered, and a significantly larger proportion of them hydrolyzed esculin and gelatin. Only 4% of the subsurface isolates fermented glucose, even though 82% of them could use it aerobically. l-Malate and d-gluconate were utilized by at least 75% of the subsurface isolates, and seven other carbon sources were used by at least 40% of them. Subsurface isolates from different geological formations (depths) and, to a lesser extent, from the same geological formation at different boreholes differed distinctly in their group responses to certain physiological tests. Moreover, sediments from different depths and boreholes contained physiologically distinct types of bacteria. Thus, considerable bacterial diversity was observed in coastal plain subsurface sediments, even within defined geological formations. PMID- 16347903 TI - Physical modeling of the composting ecosystem. AB - A composting physical model with an experimental chamber with a working volume of 14 x 10 cm (0.5 ft) was designed to avoid exaggerated conductive heat loss resulting from, relative to field-scale piles, a disproportionately large outer surface-area-to-volume ratio. In the physical model, conductive flux (rate of heat flow through chamber surfaces) was made constant and slight through a combination of insulation and temperature control of the surrounding air. This control was based on the instantaneous conductive flux, as calculated from temperature differentials via a conductive heat flow model. An experiment was performed over a 10-day period in which control of the composting process was based on ventilative heat removal in reference to a microbially favorable temperature ceiling (temperature feedback). By using the conduction control system (surrounding air temperature controlled), 2.4% of the total heat evolved from the chamber was through conduction, whereas the remainder was through the ventilative mechanisms of the latent heat of vaporization and the sensible temperature increase of air. By comparison, with insulation alone (the conduction control system was not used) conduction accounted for 33.5% of the total heat evolved. This difference in conduction resulted in substantial behavioral differences with respect to the temperature of the composting matrix and the amount of water removed. By emphasizing the slight conduction system (2.4% of total heat flow) as being a better representative of field conditions, a comparison was made between composting system behavior in the laboratory physical model and field-scale piles described in earlier reports. Numerous behavioral patterns were qualitatively similar in the laboratory and field (e.g., temperature gradient, O(2) content, and water removal). It was concluded that field-scale composting system behavior can be simulated reasonably faithfully in the physical model. PMID- 16347904 TI - Competitive Inhibition of Ferrous Iron Oxidation by Thiobacillus ferrooxidans by Increasing Concentrations of Cells. AB - The oxidation of ferrous iron (Fe) to ferric iron (Fe) with dioxygen (O(2)) by various strains of Thiobacillus ferrooxidans was studied by measuring the rate of O(2) consumption at various Fe concentrations and cell concentrations. The apparent K(m) values for Fe remained constant at different cell concentrations of laboratory strains ATCC 13661 and ATCC 19859 but increased with increasing cell concentrations of mine isolates SM-4 and SM-5. The latter results are explained by the competitive inhibition of the Fe-binding site of a cell by other cells in the reaction mixture. Possible mechanisms involving cell surface properties are discussed. PMID- 16347905 TI - Effect of growth rate and starvation-survival on the viability and stability of a psychrophilic marine bacterium. AB - Cell populations of the marine bacterium ANT-300, from either batch or continuous culture with dilution rates ranging from D = 0.015 h to D = 0.200 h, were monitored for viability, direct counts, and optical density for 98 days under starvation conditions. Three stages of starvation survival were observed for each of the cell populations. Although direct counts remained at 2 x 10 to 3 x 10 cells ml throughout the starvation period, large fluctuations occurred in cell viability during stage 1 (0 to 14 days) of starvation survival. Stage 2 (14 to 70 days) involved an overall decrease in viability for each of the cell populations; the rate of viability loss was dependent upon the growth rate. Cell viability stabilized at approximately 0.3% of the direct count in stage 3 (70 to 98 days). Long-term starvation corresponded to the prolongation of stage 3 starvation survival. Cell volumes for each of the cell populations decreased with the length of the starvation period. However, the cell volume of starved cells was also dependent more on growth rate than on the length of the time starved. We hypothesize that the cell population with the slowest growth rate is most closely representative of cells found in the oligotrophic marine environment. PMID- 16347906 TI - Colonial and Cellular Polymorphism in Xenorhabdus luminescens. AB - A highly polymorphic Xenorhabdus luminescens strain was isolated. The primary form of X. luminescens was luminescent and nonswarming and produced a yellow pigment and antimicrobial substances. The primary form generated a secondary form that had a distinct orange pigmentation, was weakly luminescent, and did not produce antimicrobial substances. Both the primary and secondary forms generated a set of colony variants at frequencies that exceeded normal rates for spontaneous mutation. The variant forms include nonswarming and swarming forms that formed large colonies and a small-colony (SC) form. The primary and secondary forms generated their SC forms at frequencies of between 1 and 14% and 1 and 2%, respectively. The SC forms were distinct from their parental primary and secondary forms in colony and cellular morphology and in protein composition. The cellular morphology and protein patterns of the nonswarming and swarming colony variants were all very similar. The DNA fingerprints of all forms were similar. Each SC-form colony reverted at high frequency to the form from which it was derived. The proportion of parental-type cells in the SC-form colonies varied with age, with young colonies containing as few as 0.0002% parental-type cells. The primary-to-secondary switch was stable, but all the other colony forms were able to switch at high frequencies to the alternative colony phenotypes. PMID- 16347907 TI - Relationship between Fungal Biomass Production and the Brightening of Hardwood Kraft Pulp by Coriolus versicolor. AB - The white-rot fungus Coriolus versicolor increased the brightness of hardwood kraft pulp by two mechanisms depending on the concentration of available nitrogen. In low-nitrogen conditions, the brightening process was a chemical effect mediated by the fungus, associated with the removal of residual lignin in the pulp; kappa number was used as an indicator of lignin concentration. A five day treatment in low-nitrogen conditions increased the brightness of hardwood kraft pulp from 36.2 to 54.5%, with a corresponding decrease in kappa number from 12.0 to 8.5, equivalent to a reduction in the lignin concentration from ca. 2.0% (wt/wt) to ca. 1.4% (wt/wt). Under these conditions, we concluded that the brightening of the pulp was a secondary metabolic event initiated after the depletion of available nitrogen. This method of brightening has been described as bleaching or biobleaching. By contrast, in high-nitrogen conditions, the brightening was a physical effect associated with the dilution of the dark pulp fibers by the relatively high levels of brighter fungal mycelium produced. Since this method of brightening was not evidently associated with lignin removal, it cannot be described as bleaching. In pulp samples brightened in high-nitrogen conditions, as brightness increased, there was a corresponding increase in kappa number. This observation was explained by the consumption of potassium permanganate by the fungal mycelium, which interfered with kappa number determinations at high fungal biomass levels. PMID- 16347908 TI - Extraction of Zinc from Industrial Waste by a Penicillium sp. AB - Zinc was extracted from a filter residue of a copper works (58.6% zinc) by a Penicillium sp. isolated from a metal-containing location. By isotachophoresis citric acid was identified as the leaching agent. Citrate was only formed when the leaching substrate was present. This production of citrate was different in several ways from that achieved by Aspergillus niger: glucose was utilized before fructose; the initial concentration of zinc was 50 to 500 times higher than usual in citrate fermentations with A. niger; citrate production stopped when 80 to 90% of the zinc was leached, although sufficient sugar for further synthesis was still present; and in synthetic media citrate production by A. niger needs an acidic environment (pH 2), while the formation of citric acid by Penicillium sp. occurred in a pH range of 7 to 4. Tests with different concentrations of waste material (0.5, 2.5, and 5%) showed that the highest yield of solubilized zinc occurred with a 2.5% substrate (93% zinc extracted after 13 days). PMID- 16347909 TI - Production of Major Extracellular Enzymes during Lignocellulose Degradation by Two Streptomycetes in Agitated Submerged Culture. AB - Streptomyces viridosporus T7A and S. badius 252 were grown in 1 to 2% (wt/vol) slurry cultures with mineral salts solution containing 0.6% yeast extract and 100/200 mesh ground and extracted corn lignocellulose at 37 degrees C. Enzyme activities rapidly increased in the first 3 to 4 days and then declined and remained at a relatively constant level. Concentrations of endoglucanase and xylanase produced by S. badius were lower than those produced by S. viridosporus. However, the lignin-peroxidase peak concentration was threefold higher than with S. viridosporus and was obtained at 9 to 10 days of incubation. By polyacrylamide gel analysis, it was determined that peroxidases from both species consisted of four enzymes, with only one, the lignin peroxidase, having high activity. A culture pH of 8.5 was preferable for lignocellulose degradation by S. badius. PMID- 16347910 TI - Solutions to problems in enumerating sediment bacteria by direct counts. AB - We examined the effect of different sediment types on the staining effectiveness of the fluorochrome DAPI (4'-6-diamidino-2-phenylindole dihydrochloride) over a wide range of concentrations and on the masking effect of sediment particles on DAPI-stained bacteria. Sediment type greatly affects the staining efficiency of DAPI, and most published studies seem to have underestimated bacterial abundances by using suboptimal concentrations of the fluorochrome. A DAPI concentration of 5 mug ml is required to effectively stain the bacteria in most sediments that can be sampled with a gravity corer. When the sediments are diluted 687 times (a dilution factor similar to those most often used in the literature), sediment particle masking of stained bacteria is highly variable for different sediment types. By using a measure of turbidity (A(750)) to indicate masking and the quartz-corrected water content as a measure of the initial (in situ) dilution of each sediment type, it becomes possible to show a linear relationship between masking and the integrated (initial x experimental) dilution of various sediments. This relationship allows the development of a correction procedure for masking which makes accurate and unbiased counts possible. Data so obtained show a strong relationship between bacteria (cells per milliliter of fresh sediment) and sediment organic matter (grams [dry weight] per milliliter of fresh sediment), one that is not discernable without the correction. The proposed method of staining and correction for sediment masking provides the basis for a standardized interpretation of sediment bacterial counts. PMID- 16347911 TI - Microbial transformation of the tetrazolinone herbicide f5231. AB - Microbial transformation of the tetrazolinone herbicide F5231 was accomplished with the filamentous fungus Absidia pseudocylindrospora Hesseltine et Ellis (ATCC 24169). The fungus converted the herbicide to six metabolites which were identified spectrally by mass, infrared, and nuclear magnetic resonance spectroscopy. PMID- 16347912 TI - Microzonation of denitrification activity in stream sediments as studied with a combined oxygen and nitrous oxide microsensor. AB - Microzonation of denitrification was studied in stream sediments by a combined O(2) and N(2)O microsensor technique. O(2) and N(2)O concentration profiles were recorded simultaneously in intact sediment cores in which C(2)H(2) was added to inhibit N(2)O reduction in denitrification. The N(2)O profiles were used to obtain high-resolution profiles of denitrification activity and NO(3) distribution in the sediments. O(2) penetrated about 1 mm into the dark-incubated sediments, and denitrification was largely restricted to a thin anoxic layer immediately below that. With 115 muM NO(3) in the water phase, denitrification was limited to a narrow zone from 0.7 to 1.4 mm in depth, and total activity was 34 nmol of N cm h. With 1,250 muM NO(3) in the water, the denitrification zone was extended to a layer from 0.9 to 4.8 mm in depth, and total activity increased to 124 nmol of N cm h. Within most of the activity zone, denitrification was not dependent on the NO(3) concentration and the apparent K(m) for NO(3) was less than 10 muM. Denitrification was the only NO(3)-consuming process in the dark incubated stream sediment. Even in the presence of C(2)H(2), a significant N(2)O reduction (up to 30% of the total N(2)O production) occurred in the reduced, NO(3)-free layers below the denitrification zone. This effect must be corrected for during use of the conventional C(2)H(2) inhibition technique. PMID- 16347913 TI - Competition by Bradyrhizobium Strains for Nodulation of the Nonlegume Parasponia andersonii. AB - Bradyrhizobium strains isolated from the nonlegume Parasponia spp. formed a group of strains that were highly competitive for nodulation of P. andersonii when paired with strains isolated from legumes. Strains from legumes, including those of similar effectiveness to NGR231 and CP283, were not able to form nodules as single occupants on P. andersonii in the presence of Parasponia strains. However, NGR86, an isolate from Macroptilium lathyroides, jointly occupied one-third of the nodules formed with each of the three strains isolated from Parasponia spp. Time taken for nodules to appear may have influenced the outcome of competition, since CP283 and all isolates from legumes were slow to nodulate P. andersonii. Among the Parasponia strains, competitiveness for nodulation of P. andersonii was not associated with effectiveness of nitrogen fixation. The highly effective strain CP299 was a poor competitor when paired with the least effective strain NGR231. CP283 was the least competitive of the Parasponia strains but was still able to dominate nodules when paired with legume isolates. Dual occupancy was high, up to 67% when the inoculum contained CP299 and CP273. Both the Muc and Muc types of CP283 form a symbiosis of similar effectiveness and were similarly competitive at high inoculation densities, but the Muc form was more competitive at low inoculum densities. Both forms frequently occupied the same nodule. Bradyrhizobium strains isolated from Parasponia spp. may have specific genetic information that favor their ability to competitively and effectively infect plants in the genus Parasponia (Ulmaceae) outside the Leguminosae. PMID- 16347914 TI - Metal Interactions with Microbial Biofilms in Acidic and Neutral pH Environments. AB - Microbial biofilms were grown on strips of epoxy-impregnated filter paper submerged at four sites in water contaminated with metals from mine wastes. At two sample stations, the water was acidic (pH 3.1); the other sites were in a lake restored to a near neutral pH level by application of a crushed limestone slurry. During a 17-week study period, planktonic bacterial counts increased from 10 to 10 CFU/ml at all sites. Biofilm counts increased rapidly over the first 5 weeks and then leveled to 10 CFU/cm in the neutral pH system and 10 CFU/cm at the acidic sites. In each case, the biofilms bound Mn, Fe, Ni, and Cu in excess of the amounts adsorbed by control strips covered with nylon filters (pore size, 0.22 mum) to exclude microbial growth; Co bound under neutral conditions but not under acidic conditions. Conditional adsorption capacity constants, obtained graphically from the data, showed that biofilm metal uptake at a neutral pH level was enhanced by up to 12 orders of magnitude over acidic conditions. Similarly, adsorption strength values were usually higher at elevated pH levels. In thin sections of the biofilms, encapsulated bacterial cells were commonly found enmeshed together in microcolonies. The extracellular polymers often contained iron oxide precipitates which generated weak electron diffraction patterns with characteristic reflections for ferrihydrite (Fe(2)O(3) . H(2)O) at d equaling 0.15 and 0.25 nm. At neutral pH levels, these deposits incorporated trace amounts of Si and exhibited a granular morphology, whereas acicular crystalloids containing S developed under acidic conditions. PMID- 16347915 TI - Temperature Compensation in Methanosarcina barkeri by Modulation of Hydrogen and Acetate Affinity. AB - The affinity of Methanosarcina barkeri 227 for acetate and hydrogen at different incubation temperatures was investigated. Increasing the temperature from 20 to 37 degrees C resulted in a 4.5-fold increase in K(m) for acetate and a 4.8-fold increase for hydrogen. The corresponding increase in V(max) for acetate was 8.3 fold (5.4-fold for hydrogen). This response implied a decrease in the temperature coefficient (Q(10)) and hence a decrease in the temperature dependency as a function of decreasing substrate concentration. PMID- 16347916 TI - Uptake and incorporation of thymidine by bacterial isolates from an upwelling environment. AB - Thymidine uptake and incorporation by marine bacterial isolates from an upwelling environment were studied. Of 17 isolates each from upwelled and downwelled water, 1 and 6 isolates, respectively, were found to be negative for [H]thymidine incorporation at a substrate concentration of 19 muM. Strains lacking the ability to take up thymidine were not confined to one genus. The measurable rates of uptake and incorporation by the 34 isolates varied greatly. Studies carried out using starved Vibrio, Pseudomonas, and Cytophaga cells showed that these isolates transported and incorporated thymidine after periods of as long as 5 weeks of nutrient deprivation. This occurred in the absence of any other exogenously supplied nutrients. Overall, these results indicate that not all marine bacteria take up thymidine and that those that do incorporate the nucleoside may do so at very different rates. The assumption that only actively growing or dividing cells incorporate thymidine must be viewed with caution. PMID- 16347917 TI - Use of the Most-Probable-Number Technique To Detect Polymyxa betae (Plasmodiophoromycetes) in Soil. AB - The fungus Polymyxa betae is an obligate parasite of the roots of many plants of the family Chenopodiaceae. In the sugar beet, it acts as a vector of beet necrotic yellow vein virus, the agent of a serious disease known as rhizomania. With indirect methods of analysis, such as bioassay, one can establish only the presence or absence, but not the quantity, of P. betae in soil. A new method based on the technique of the most probable number (MPN) of infective units of P. betae present in the soil was developed on the basis of the biological characteristics of this microorganism. Compared with traditional bioassay methods, the MPN method is suitable for determining the contamination level of P. betae in a soil, and it appears promising for the routine analysis of many soil samples, whether they were affected by rhizomania or presumed noninfested. The instrumentation designed especially for the recovery of viable P. betae from soil with the MPN technique is made from commercially available materials, results in a saving of space during sample incubation, and permits this method to be used for any laboratory analysis. PMID- 16347918 TI - Separation of Protein Crystals from Spores of Bacillus thuringiensis by Ludox Gradient Centrifugation. AB - A method is described for the purification of Bacillus thuringiensis protein crystals by Ludox gradient centrifugation. This method is simple, inexpensive, fast, and efficient compared with other techniques. It has been successfully used to purify and characterize the protein crystals from several B. thuringiensis strains. PMID- 16347919 TI - Biodegradation of Natural and Synthetic Humic Acids by the White Rot Fungus Phanerochaete chrysosporium. AB - Biodegradation of natural and synthetic (melanoidin) humic acids by Phanerochaete chrysosporium BKM-F 1767 was demonstrated by decolorization in batch culture, reduction in molecular weight, and CO(2) production from labeled melanoidin. This biodegradation occurred during secondary metabolism of the fungus in nitrogen limited cultures; experimental results suggest that all or a part of the lignin degrading system of BKM-F 1767 plays a part in biodegradation. PMID- 16347920 TI - Styrofoam cup-membrane assembly for studying microorganism-root interactions. AB - An assembly consisting of Styrofoam cups with membranes of varying porosities was developed to study microorganism-root interactions. The assembly permitted uniform distribution of a bacterium in soil and was simple, easy to use, and disposable. In tests with the bacterium Pseudomonas solanacearum, little difference in P. solanacearum survival was observed in the rhizosphere or nonrhizosphere of tomato. PMID- 16347921 TI - Regulation of Glycolytic Flux and Ethanol Production in Saccharomyces cerevisiae: Effects of Intracellular Adenine Nucleotide Concentrations on the In Vitro Activities of Hexokinase, Phosphofructokinase, Phosphoglycerate Kinase, and Pyruvate Kinase. AB - The progressive decline in the glycolytic activity of Saccharomyces cerevisiae during batch fermentation is accompanied by changes in adenine nucleotide pools. The relative activities of four glycolytic enzymes were examined in vitro in the presence of nucleotide concentrations equivalent to intracellular pools. Phosphofructokinase and pyruvate kinase were not inhibited. Phosphoglycerate kinase was inhibited by AMP but was judged unlikely to be of physiological consequence owing to enzyme abundance. Both isoenzymes of hexokinase were strongly inhibited by AMP. The degree of hexokinase inhibition was sufficient to account for the observed decline in glycolytic activity during batch fermentation. PMID- 16347922 TI - Variation in Deoxynivalenol, 15-Acetyldeoxynivalenol, 3-Acetyldeoxynivalenol, and Zearalenone Production by Fusarium graminearum Isolates. AB - Of 88 isolates of Fusarium graminearum collected from soil or cereals in the United States, 49 produced 15-acetyldeoxynivalenol (15-ADON) as the major isomer; one produced 3-acetyldeoxynivalenol (3-ADON). A total of 26 isolates collected from cereals or soil in Australia, New Zealand, Norway, China, and Poland were used for comparison. Of these, 15 produced 3-ADON as the major isomer and 2 produced 15-ADON. PMID- 16347923 TI - Synthetic Medium for Acetobacter xylinum That Can Be Used for Isolation of Auxotrophic Mutants and Study of Cellulose Biosynthesis. AB - Acetobacter xylinum is a bacterium that can synthesize cellulose when grown in an undefined medium containing glucose. We developed a defined minimal medium for A. xylinum that contains 1% glucose, 0.1% NH(4)Cl, 0.115% citric acid, 0.33% Na(2)HPO(4), 0.01% KCl, 0.025% MgSO(4). 7H(2)O, and 7.5 mg of nicotinamide per liter which both allows cellulose synthesis and can be used to isolate a variety of auxotrophic mutants. PMID- 16347924 TI - Use of robotics to dispense culture media. AB - Liquid and agar-containing media can be aseptically dispensed by a laboratory robot. Five errors occurred during the filling of 700 tubes with medium; four of these were due to imperfections of the screw-threaded caps of the culture tubes. The mean volume dispensed was 14.8 ml (standard deviation, 0.5; n = 50) when the system was set to deliver 15 ml. PMID- 16347925 TI - Hyperproduction of Poly-beta-Hydroxybutyrate during Exponential Growth of Azotobacter vinelandii UWD. AB - The transformation of Azotobacter vinelandii UW with A. vinelandii 113 DNA resulted in the formation of rifampin-resistant colonies, 13% of which also inherited a previously unrecognized mutation in the respiratory NADH oxidase. These transformants produced colonies with a white-sectored phenotype after prolonged incubation. Cells from these sectors were separated and purified by streaking and were named UWD. The dense white phenotype was due to the production of a large amount of poly-beta-hydroxybutyrate during the exponential growth of strain UWD. The polymer accounted for 65 or 75% of the cell dry weight after 24 h of incubation of cultures containing glucose and either ammonium acetate or N(2), respectively, as the nitrogen source. Under the same conditions, strain UW cells contained 22 to 25% poly-beta-hydroxybutyrate, but O(2)-limited growth was required for these optimal production values. Polymer production was not dependent on O(2) limitation in strain UWD, but the efficiency of conversion of glucose to poly-beta-hydroxybutyrate was enhanced in O(2)-limited cultures. Conversion efficiencies were >0.25 and 0.33 mg of poly-beta-hydroxybutyrate per mg of glucose consumed under vigorous- and low-aeration conditions, respectively, compared with an efficiency of 0.05 achieved by strain UW. Strain UWD, therefore, appeared to from poly-beta-hydroxybutyrate under novel conditions, which may be useful in designing new methods for the industrial production of biodegradable plastics. PMID- 16347926 TI - Effect of Surface-Active Pseudomonas spp. on Leaf Wettability. AB - Different strains of Pseudomonas putida and P. fluorescens isolated from the rhizosphere and phyllosphere were tested for surface activity in droplet cultures on polystyrene. Droplets of 6 of the 12 wild types tested spread over the surface during incubation, and these strains were considered surface active; strains not showing this reaction were considered non-surface active. Similar reactions were observed on pieces of wheat leaves. Supernatants from centrifuged broth cultures behaved like droplets of suspensions in broth; exposure to 100 degrees C destroyed the activity. Average contact angles of the supernatants of surface active and non-surface-active strains on polystyrene were 24 degrees and 72 degrees , respectively. The minimal surface tension of supernatants of the surface-active strains was about 46 mN/m, whereas that of the non-surface-active strains was 64 mN/m (estimations from Zisman plots). After 6 days of incubation, wheat flag leaves sprayed with a dilute suspension of a surface-active strain of P. putida (WCS 358RR) showed a significant increase in leaf wettability, which was determined by contact angle measurements. Increasing the initial concentration of bacteria and the amount of nutrients in the inoculum sprayed on leaves reduced the contact angles from 138 degrees on leaves treated with antibiotics (control) to 43 degrees on leaves treated with surface-active bacteria. A closely related strain with no surface activity on polystyrene did not affect leaf wettability, although it was present in densities similar to those of the surface-active strain. Nutrients alone could occasionally also increase leaf wettability, apparently by stimulating naturally occurring surface active bacteria. When estimating densities of Pseudomonas spp. underneath droplets with low contact angles, it appeared that populations on leaves treated with a surface-active strain could vary from about 10 to 10 CFU cm, suggesting that the surface effect may be prolonged after a decline of the population. The possible ecological implications are discussed. PMID- 16347927 TI - Isolation, growth characteristics, and long-term storage of fungi cultivated by attine ants. AB - Seven pure-culture strains of fungi cultivated by attine ants (ant-garden fungi) were isolated from locally maintained leaf-cutting ant colonies. An ant-garden fungus strain obtained from an Atta cephalotes colony, when offered to ants of the colony from which the fungus was isolated, was accepted as their own. Young fungus cultures were harvested and incorporated into the fungus garden, and cultures of intermediate age were used to begin a new fungus garden; old cultures were simply harvested. To facilitate further research on this fungus, growth characteristics of the different isolates were studied under a variety of conditions. They grew better at 24 degrees C than at 30 degrees C, and growth did not occur at an incubation temperature of 37 degrees C. In a broth culture medium, growth was enhanced by aeration of the culture and by addition of yeast extract, olive oil, sesame oil, peanut oil, soybean oil, corn oil, sunflower oil, cottonseed oil, walnut oil, safflower oil, or mineral oil. Glycerol did not noticeably affect growth, but Tween 80 inhibited growth. These fungi were extremely sensitive to cycloheximide, growth being totally inhibited at cycloheximide concentrations ranging from 0.4 to 4.0 mug/ml. To date, the ant garden fungus isolates have remained viable in long-term mineral oil-overlay storage cultures for up to 4 years. PMID- 16347928 TI - Biodegradation of asphalt cement-20 by aerobic bacteria. AB - Seven gram-negative, aerobic bacteria were isolated from a mixed culture enriched for asphalt-degrading bacteria. The predominant genera of these isolates were Pseudomonas, Acinetobacter, Alcaligenes, Flavimonas, and Flavobacterium. The mixed culture preferentially degraded the saturate and naphthene aromatic fractions of asphalt cement-20. A residue remained on the surface which was resistant to biodegradation and protected the underlying asphalt from biodegradation. The most potent asphalt-degrading bacterium, Acinetobacter calcoaceticus NAV2, excretes an emulsifier which is capable of emulsifying the saturate and naphthene aromatic fractions of asphalt cement-20. This emulsifier is not denatured by phenol. PMID- 16347929 TI - Growth of Neocallimastix sp. Strain R1 on Italian Ryegrass Hay: Removal of Neutral Sugars from Plant Cell Walls. AB - The anaerobic fungus Neocallimastix sp. strain R1 was grown for up to 5 days on a medium containing autoclaved Italian ryegrass hay as the carbon source. Culture supernatants and digested cell walls were harvested at 12-h intervals. Supernatants were analyzed for the fermentation products formate and acetate, and residual cell walls were analyzed for dry-matter and neutral-sugar losses. Fungal growth was accompanied by the digestion of plant cell walls and the accumulation of fermentation products in culture media. Dry-matter losses were accounted for by removal of four major neutral sugars (arabinose, galactose, glucose, and xylose) from the plant cell walls. First-order reaction kinetics could be used to describe the loss of each sugar. All cell wall sugars, including arabinose and galactose, which are not fermented by Neocallimastix sp. strain R1 were removed simultaneously. Although the rates of removal of individual sugars were similar, there were significant differences in their extents of removal: the extent of removal of arabinose exceeded that of the other three sugars, and xylose was the least digestible. This study provides the first account of simultaneous (nonpreferential) removal of neutral sugars from plant cell walls by an anaerobic fungus. Although in vitro techniques were used, these results indicate a potentially significant role for the anaerobic fungi as fiber digesters in the rumen. PMID- 16347930 TI - Characterization of bacteria that suppress rhizoctonia damping-off in bark compost media by analysis of Fatty Acid biomarkers. AB - Examination of cucumber roots (Cucumis sativus L.) grown in bark compost media and of the surrounding edaphic substrate showed profiles of polar lipid fatty acids commonly found in bacteria. The composition of fatty acids in these profiles differed significantly between roots grown in a medium naturally suppressive to Rhizoctonia damping-off and roots from a conducive medium. Cucumber roots from the suppressive medium had higher proportions of cis-vaccenic acid (18:1 omega 7c) and the iso-branched monoenoic fatty acid i17:1 omega 8 but lower proportions of several iso- and anteiso-branched fatty acids compared with roots from the conducive medium. The concentrations of the bacterial fatty acids were significantly lower in the surrounding media. However, the suppressive and conducive growth substrates had differences in the composition of the bacterial fatty acids similar to those found between the cucumber roots proper. These results suggest major differences in bacterial community composition between suppressive and conducive systems. Fatty acid analyses were also utilized to examine the effects on bacterial community composition of root colonization by Flavobacterium balustinum 299, a biocontrol agent. The concentration of the most prominent fatty acid in this bacterium, i17:1 omega 8, was increased on roots produced from inoculated seeds in a medium rendered suppressive by the treatment. This change was concomitant with a significant increase in the concentration of 18:1 omega 7c, not present in the lipids of the antagonist, indicating a shift in the microflora from a conducive to a suppressive bacterial community. PMID- 16347931 TI - Germination of Clostridium cylindrosporum Spores on Medium Containing Uric Acid. AB - Clostridium cylindrosporum spores germinated rapidly under reducing conditions when bicarbonate, uric acid, and calcium were present. Germination rates on 10 mM urate increased with increasing Ca (maximum rate at 5 mM Ca or greater). Germination rates on urate (limiting Ca) increased with increasing urate concentrations to 10 mM urate. At 10 mM Ca, germination rates reached a maximum at 1 mM urate and remained constant thereafter. Cations (Na, K, Li, and Mg), purines, purine analogs, and EDTA inhibited germination at limiting calcium concentrations but not (except for 10 mM adenine) at 10 mM Ca. Methyl viologen or formate did not inhibit germination. Germination was not observed in solutions containing xanthine, hypoxanthine, caffeine, theophylline, 6,8-dihydroxypurine, adenine, allopurinol, formate, glycine, or acetate, even though some of the purines are growth substrates. PMID- 16347932 TI - Seasonal Biotransformation of Naphthalene, Phenanthrene, and Benzo[a]pyrene in Surficial Estuarine Sediments. AB - Transformation rates of naphthalene, phenanthrene, and benzo[a]pyrene in oxidized surficial sediments of a polluted urban estuary, Boston Harbor, Mass., were determined over a period of 15 months. Three sites characterized by muddy sediments were selected to represent a >300-fold range of ambient polycyclic aromatic hydrocarbon (PAH) concentration. Transformation rates were determined by a trace-level radiolabel PAH assay which accounted for PAH mineralization, the formation of polar metabolites, residue, and recovered parental PAHs in sediment slurries. Transformation rates of the model PAHs increased with increasing ambient PAH concentrations. However, turnover times for a given PAH were similar at all sites. The turnover times were as follows: naphthalene, 13.2 to 20.1 days; phenanthrene, 7.9 to 19.8 days, and benzo[a]pyrene, 53.7 to 82.3 days. At specific sites, rates were significantly affected by salinity, occasionally affected by temperature, but not affected by pH over the course of the study. Seasonal patterns of mineralization were observed for each of the PAHs at all sites. The timing of seasonal maxima of PAH mineralization varied from site to site. Seasonal potential heterotrophic activities as measured by acetate and glutamate mineralization rates did not always coincide with PAH mineralization maxima and minima, suggesting that the two processes are uncoupled in estuarine sediments. PMID- 16347933 TI - Volatilization of Selenium by Alternaria alternata. AB - Seleniferous water continues to be a serious problem to wildlife in the central valley of California. Water samples collected from Kesterson Reservoir, Peck Ranch, and Lost Hills evaporation pond facilities contained between 0.005 and 5 mg of Se per liter. The objective of this study was to isolate Se-methylating organisms in evaporation pond water and to assess, through enrichment and manipulation of their optimal growth parameters, the environmental factors which govern microbial Se methylation. Alternaria alternata was isolated as an active Se-methylating organism. The volatile product was identified as dimethylselenide. The effects of pH, temperature, Se substrates, and methyl donors on the ability of A. alternata to methylate Se were investigated in liquid medium containing 100 mg of Se per liter. The optimum pH and temperature for methylation were 6.5 and 30 degrees C, respectively. Selenate and selenite were methylated more rapidly than selenium sulfide and various organic Se compounds (6-selenoguanosine, 6 selenoinosine, seleno-dl-methionine, and 6-selenopurine). l-Methionine and methyl cobalamine (0.1 muM) stimulated dimethylselenide production. This study demonstrates that Se-methylating organisms are present in evaporation pond water and are capable of liberating substantial quantities of Se in the volatile dimethylselenide form. By determining the optimum environmental conditions which stimulate volatilization, it may be possible to design a way to remove Se from seleniferous water in situ. PMID- 16347934 TI - Isolation and Ultrastructure of the Flagella of Methanococcus thermolithotrophicus and Methanospirillum hungatei. AB - The flagella of the archaebacteria Methanococcus thermolithotrophicus and Methanospirillum hungatei enter the cells in regions with ultrastructure resembling that of the polar organelles found in a variety of eubacteria. Flagella of both organisms consist of a filament, a hook, and a basal body with two rings similar to those of gram-positive eubacteria. The integrity of the flagella of M. thermolithotrophicus is lost in the absence of high salt concentrations, and those of both organisms are unstable at high pH. The flagellar filaments of M. hungatei are composed of two flagellins of 24 and 26 kilodaltons. PMID- 16347935 TI - Role of DNA Superhelicity in Regulation of Bacteroid-Associated Functions of Bradyrhizobium sp. Strain 32H1. AB - Bradyrhizobium sp. strain 32H1 cells express a number of bacteroid-associated functions and repress some functions related to the free-living state when grown ex planta under conditions of low (0.2%) oxygen tension and relatively high levels (>8 mM) of medium K. Expression of the bacteroid-associated phenotype was blocked by the DNA gyrase inhibitor novobiocin. Because the degree of negative supercoiling of DNA is the result of the activities of both DNA gyrase and topoisomerase I, we measured these enzymes in cells grown under nitrogen-fixing (low O(2), high K) and non-nitrogen-fixing conditions (low O(2), low [50 muM] K or high O(2), high K). Lower topoisomerase I activities were seen in extracts from nitrogen-fixing cells than in those from non-nitrogen-fixing cells. In contrast, DNA gyrase levels were lower in high-O(2)-grown cells than under the other conditions tested. These differences are consistent with an increase in DNA superhelicity associated with growth under low-O(2), high-K conditions. A spontaneous mutant resistant to the DNA gyrase inhibitor ciprofloxacin was found to be constitutive with respect to the K requirement, because it expressed the bacteroid-associated phenotype when grown under low-O(2), low-K conditions. The mutant cells gave rise to effective nodules on Macroptilium atropurpureum and possessed the low topoisomerase I activities and high DNA gyrase levels of low O(2)-, high-K-grown wild-type cells. Our data suggest that changes in DNA supercoiling resulting from low O(2) tension and a high K concentration exert a major influence on the expression of the bacteroid-associated phenotype. PMID- 16347936 TI - Expression by Soil Bacteria of Nodulation Genes from Rhizobium leguminosarum biovar trifolii. AB - Gram-negative, rod-shaped bacteria from the soil of white clover-ryegrass pastures were screened for their ability to nodulate white clover (Trifolium repens) cultivar Grasslands Huia and for DNA homology with genomic DNA from Rhizobium leguminosarum biovar trifolii ICMP2668 (NZP582). Of these strains, 3.2% were able to hybridize with strain ICMP2668 and nodulate white clover and approximately 19% hybridized but were unable to nodulate. Strains which nodulated but did not hybridize with strain ICMP2668 were not detected. DNA from R. leguminosarum biovar trifolii (strain PN165) cured of its symbiotic (Sym) plasmid and a specific nod probe were used to show that the relationship observed was usually due to chromosomal homology. Plasmid pPN1, a cointegrate of the broad host-range plasmid R68.45 and a symbiotic plasmid pRtr514a, was transferred by conjugation to representative strains of nonnodulating, gram-negative, rod-shaped soil bacteria. Transconjugants which formed nodules were obtained from 6 of 18 (33%) strains whose DNA hybridized with that of PN165 and 1 of 9 (11%) strains containing DNA which did not hybridize with that of PN165. The presence and location of R68.45 and nod genes was confirmed in transconjugants from three of the strains which formed nodules. Similarly, a pLAFR1 cosmid containing nod genes from a derivative of R. leguminosarum biovar trifolii NZP514 formed nodules when transferred to soil bacteria. PMID- 16347937 TI - Bacterial Colonization of Cod (Gadus morhua L.) and Halibut (Hippoglossus hippoglossus) Eggs in Marine Aquaculture. AB - Aquaculture has brought about increased interest in mass production of marine fish larvae. Problems such as poor egg quality and mass mortality of fish larvae have been prevalent. The intensive incubation techniques that often result in bacterial overgrowth on fish eggs could affect the commensal relationship between the indigenous microflora and opportunistic pathogens and subsequently hamper egg development, hatching, larval health, and ongrowth. Little information about the adherent microflora on fish eggs is available, and the present study was undertaken to describe the microbial ecology during egg development and hatching of two fish species of potential commercial importance in marine aquaculture. Attachment and development of the bacterial flora on cod (Gadus morhua L.) eggs from fertilization until hatching was studied by scanning electron microscopy. The adherent microflora on cod (G. morhua L.) and halibut (Hippoglossus hippoglossus) eggs during incubation was characterized and grouped by cluster analysis. Marked bacterial growth could be demonstrated 2 h after fertilization, and at hatching eggs were heavily overgrown. Members of the genera Pseudomonas, Alteromonas, Aeromonas, and Flavobacterium were found to dominate on the surface of both cod and halibut eggs. The filamentous bacterium Leucothrix mucor was found on eggs from both species. While growth of L. mucor on halibut eggs was sparse, cod eggs with a hairy appearance due to overgrowth by this bacterium close to hatching were frequently observed. Vibrio fischeri could be detected on cod eggs only, and pathogenic vibrios were not detected. Members of the genera Moraxella and Alcaligenes were found only on halibut eggs. Caulobacter and Seliberia spp. were observed attached to eggs dissected from cod ovaries under sterile conditions, indicating the presence of these bacteria in ovaries before spawning. Adherent strains did not demonstrate antibiotic resistance above a normal level. Attempts to regulate the egg microflora by incubation of gnotobiotic eggs with defined antibiotic-producing strains did not result in persistent protection against subsequent colonization by the microflora of the incubator. PMID- 16347938 TI - Selection by Anion-Exchange Chromatography of Exopolysaccharide Mutants of the Cyanobacterium Synechocystis Strain PCC 6803. AB - The degree of retention of whole cells of Synechocystis strain PCC 6803 on DEAE cellulose columns was shown to depend on their content of exopolysaccharides, which are at least in part responsible for the external negative charge of the cells. This feature was used for the isolation of mutants modified in the apparent viscosity caused by these macromolecular constituents. When a wild-type suspension was loaded onto a DE52 column, the cells eluting in the two extreme fractions of a 0 to 5 M NaCl step gradient represented 10 to 10 of the total eluted population. The accuracy of the procedure was established through the analysis of four clones: Suc(0)32 and Suc(0)65 (0 M) and Suc(5)64A and Suc(5)61 (5 M). The decreased viscosity of the exopolymers of the two 0 M clones, which appeared identical, could be related to the production of molecules less charged in uronic acids and more readily liberated from the cells. The two 5 M clones exhibited a lower sedimentation velocity, correlating with either a 60% increase in uronic acid and a doubling of the specific viscosity of the exopolysaccharides [clone Suc(5)64A] or a doubling of the per-cell production of polymers otherwise identical to those from wild-type cells [clone Suc(5)61]. PMID- 16347939 TI - Detection of lignin peroxidase and xylanase by immunocytochemical labeling in wood decayed by basidiomycetes. AB - The white rot fungi used in this study caused two different forms of degradation. Phanerochaete chrysosporium, strain BKM-F-1767, and Phellinus pini caused a preferential removal of lignin from birch wood, whereas Trametes (Coriolus) versicolor caused a nonselective attack of all cell wall components. Use of polyclonal antisera to H8 lignin peroxidase and monoclonal antisera to H2 lignin peroxidase followed by immunogold labeling with protein A-gold or protein G-gold, respectively, showed lignin peroxidase extra-and intracellularly to fungal hyphae and within the delignified cell walls after 12 weeks of laboratory decay. Lignin peroxidase was localized at sites within the cell wall where electron-dense areas of the lignified cell wall layers remained. In wood decayed by Trametes versicolor, lignin peroxidase was located primarily along the surface of eroded cell walls. No lignin peroxidase was evident in brown-rotted wood, but slight labeling occurred within hyphal cells. Use of polyclonal antisera to xylanase followed by immunogold labeling showed intense labeling on fungal hyphae and surrounding slime layers and within the woody cell wall, where evidence of degradation was apparent. Colloidal-gold-labeled xylanase was prevalent in wood decayed by all fungi used in this study. Areas of the wood with early stages of cell wall decay had the greatest concentration of gold particles, while little labeling occurred in cells in advanced stages of decay by brown or white rot fungi. PMID- 16347940 TI - Anaerobic Degradation of Chloroaromatic Compounds in Aquatic Sediments under a Variety of Enrichment Conditions. AB - Anaerobic degradation of monochlorophenols and monochlorobenzoates in a variety of aquatic sediments was compared under four enrichment conditions. A broader range of compounds was degraded in enrichments inoculated with sediment exposed to industrial effluents. Degradation of chloroaromatic compounds was observed most often in methanogenic enrichments and in enrichments amended with 1 mM bromoethane sulfonic acid. Degradation was observed least often in enrichments with added nitrate or sulfate. The presence of 10 mM bromoethane sulfonic acid prevented or inhibited degradation of most compounds tested. Primary enrichments in which KNO(3) was periodically replenished to maintain enrichment characteristics degraded chlorobenzoates, but not chlorophenols. In contrast, primary enrichments in which Na(2)SO(4) was periodically replenished failed to degrade any chloroaromatic compounds. Upon transfer to fresh medium, none of the sulfate enrichments required the presence of Na(2)SO(4) for degradation, while only two nitrate enrichments required the presence of KNO(3) for degradation. As a class of compounds, chlorophenols were degraded more readily than chlorobenzoates. However, as individual compounds 3-chlorobenzoate, 2 chlorophenol, and 3-chlorophenol degradation was observed most often and with an equal frequency. Within the chlorophenol class, the relative order of degradability was ortho > meta > para, while that of chlorobenzoates was meta > ortho > para, In laboratory transfers, 2-chlorobenzoate, 3-chlorobenzoate, and 2 chlorophenol degradation was most easily maintained, while degradation of para chlorinated compounds was very difficult to maintain. PMID- 16347941 TI - Characterization of anaerobic dechlorinating consortia derived from aquatic sediments. AB - Four methanogenic consortia which degraded 2-chlorophenol, 3-chlorophenol, 2 chlorobenzoate, and 3-chlorobenzoate, respectively, and one nitrate-reducing consortium which degraded 3-chlorobenzoate were characterized. Degradative activity in these consortia was maintained by laboratory transfer for over 2 years. In the methanogenic consortia, the aromatic ring was dechlorinated before mineralization to methane and carbon dioxide. After dechlorination, the chlorophenol consortia converted phenol to benzoate before mineralization. All methanogenic consortia degraded both phenol and benzoate. The 3-chlorophenol and 3-chlorobenzoate consortia also degraded 2-chlorophenol. No other cross acclimation to monochlorophenols or monochlorobenzoates was detected in the methanogenic consortia. The consortium which required nitrate for the degradation of 3-chlorobenzoate degraded benzoate and 4-chlorobenzoate anaerobically in the presence of KNO(3), but not in its absence. This consortium also degraded benzoate, but not 3-chlorobenzoate, aerobically. PMID- 16347942 TI - Genetically Engineered Erwinia carotovora in Aquatic Microcosms: Survival and Effects on Functional Groups of Indigenous Bacteria. AB - The survival of genetically engineered Erwinia carotovora L-864, with a kanamycin resistance gene inserted in its chromosome, was monitored in the water and sediment of aquatic microcosms. The density of genetically engineered and wild type E. carotovora strains declined at the same rate, falling in 32 days below the level of detection by viable counts. We examined the impact of the addition of genetically engineered and wild-type strains on indigenous bacteria belonging to specific functional groups important in nutrient cycling. For up to 16 days, the densities of total and proteolytic bacteria were significantly higher (P < 0.05) in microcosms inoculated with genetically engineered or wild-type E. carotovora, but by 32 days after inoculation, they had decreased to densities similar to those in control microcosms. Inoculation of genetically engineered or wild-type E. carotovora had no apparent effect on the density of amylolytic and pectolytic bacteria in water and sediment. Genetically engineered and wild-type E. carotovora did not have significantly different effects on the densities of specific functional groups of indigenous bacteria (P > 0.05). PMID- 16347943 TI - Principal-Component Analysis of the Characteristics Desirable in Baker's Yeasts. AB - Twenty-seven properties considered to be required for good bakery products were examined in 56 industrial and 2 laboratory yeast strains. The data obtained were applied to principal-component analysis, one of the multivariate statistical analyses. The first and second principal components together were extracted, and these accounted for 77.7% of the variance. The first principal component was interpreted as the glycolytic activity of yeast in dough, and the second one was interpreted as the balance of leavening abilities in sweet and flour doughs from the factor loadings. The scattergram on the two principal components was effective in grouping the 58 yeast strains used. PMID- 16347944 TI - Regulation of Trichodiene Synthase in Fusarium sporotrichioides and Gibberella pulicaris (Fusarium sambucinum). AB - The regulation of trichodiene synthase (TS) and its relationship to trichothecene biosynthesis was investigated in Fusarium sporotrichioides NRRL 3299 and Gibberella pulicaris R-6380. Cultures were analyzed for the presence of TS activity, trichothecenes, and immunodetectable TS polypeptide over a time period of 144 h. Enzyme activity increased from barely detectable to maximum levels over a period of 3 h for F. sporotrichioides, while in G. pulicaris, a steady increase was observed over 144 h. Increases in TS activity of 50-fold for F. sporotrichioides and 10-fold for G. pulicaris R-6380 preceded by several hours the detection of trichothecenes. Immunoblot analysis employing polyclonal serum specific for the enzyme from F. sporotrichioides showed that increases in the levels of TS polypeptide corresponded to the observed changes in enzyme activity for both organisms. These data indicate that the regulation of TS activity is accomplished through increases in its cellular concentration and that TS may serve as a useful indicator of trichothecene biosynthetic activity. PMID- 16347945 TI - Regulation of bacterioplankton production and cell volume in a eutrophic estuary. AB - During three periods of 16 to 25 days, bacterioplankton production, bacterial cell volume, chlorophyll a, CO(2) assimilation, and particulate organic carbon were measured in enclosures situated in the eutrophic estuary Roskilde Fjord, Denmark. The enclosures were manipulated with respect to sediment contact and contents of inorganic nutrients, planktivorous fish, and suspension-feeding bivalves. Nutrient enrichment, the presence of suspension feeders, and sediment contact induced pronounced changes in bacterial production, as well as minor changes in bacterial cell volume; however, these effects seemed to be indirect, transmitted via phytoplankton. Bacterial production, measured as [H]thymidine incorporation, closely followed changes in phytoplankton biomass and production, with time lags of 5 to 10 days. Good correlations of mean bacterioplankton production to chlorophyll a concentration and CO(2) assimilation suggested phytoplankton to be the dominating source of bacterial substrate, apparently independent of nutrient stress. Zooplankton >140 mum, bivalves, and sediment seemed to provide insignificant, if any, substrate for bacterioplankton, and benthic suspension feeders seemed not to act as direct competitors for dissolved organic carbon. The bacterioplankton mean cell volume, measured by image analysis, changed seasonally, with the smallest cells during the summer. Within each period, the bacterial cell volume correlated positively to growth rate and negatively to temperature. PMID- 16347946 TI - Novel Alterations in Plasmid DNA Associated with Aromatic Hydrocarbon Utilization by Pseudomonas putida R5-3. AB - Subcultures of Pseudomonas putida R5-3 altered their plasmid DNA content in specific ways depending on the particular aromatic hydrocarbon utilized as the sole carbon source. Two indigenous plasmids, 115 and 95 kilobases (kb) in size, were observed in R5-3A, which was derived from R5-3 by growth on minimal medium containing p-methylbenzoate as the sole carbon source. When R5-3A was transferred to medium containing m-xylene or toluene, derivative strains were obtained in which the 95-kb plasmid was lost and a new plasmid of 50 or 60 kb appeared. Reversion to the original plasmid profile of R5-3A was observed when xylene- or toluene-grown cells were returned to medium containing p-methylbenzoate. Restriction enzyme analysis and Southern blot hybridizations of total plasmid DNA indicated deletions and rearrangements of DNA restriction fragments in the derivatives maintained on m-xylene and toluene when compared with the original R5 3A. In the derivatives which retrieved the original plasmid profile, the restriction enzyme fragment pattern was identical to that in the original R5-3A, in that the fragments which were missing after growth on m-xylene or toluene were again present. Southern blot hybridizations revealed that part of the plasmid DNA lost from the original plasmid profile was integrated into the chromosomal DNA of xylene-grown R5-3B and that these plasmid fragments were associated with aromatic hydrocarbon metabolism. Hybridization with pathway-specific DNA fragments from the TOL plasmid pWWO indicated that this 95-kb plasmid contains DNA homologous to the meta-fission pathway genes. PMID- 16347947 TI - Resistance against Industrial Bacteriophages Conferred on Lactococci by Plasmid pAJ1106 and Related Plasmids. AB - Plasmid pAJ1106 and its deletion derivative, plasmid pAJ2074, conferred lactose fermenting ability (Lac) and bacteriophage resistance (Hsp) at 30 degrees C to Lac proteinase (Prt)-negative Lactococcus lactis subsp. lactis and L. lactis subsp. lactis var. diacetylactis recipient strains. An additional plasmid, pAJ331, isolated from the original source strain of pAJ1106, retained Hsp and conjugative ability without Lac. pAJ331 was conjugally transferred to two L. lactis subsp. lactis and one L. lactis subsp. cremoris starter strains. The transconjugants from such crosses acquired resistance to the phages which propagated on the parent recipient strains. Of 10 transconjugant strains carrying pAJ1106 or one of the related plasmids, 8 remained insensitive to phages through five activity test cycles in which cultures were exposed to a large number of industrial phages at incubation temperatures used in lactic casein manufacture. Three of ten strains remained phage insensitive through five cycles of a cheesemaking activity test in which cultures were exposed to approximately 80 different phages through cheesemaking temperatures. Three phages which propagated on transconjugant strains during cheesemaking activity tests were studied in detail. Two were similar (prolate) in morphology and by DNA homology to phages which were shown to be sensitive to the plasmid-encoded phage resistance mechanism. The third phage was a long-tailed, small isometric phage of a type rarely found in New Zealand cheese wheys. The phage resistance mechanism was partially inactivated in most strains at 37 degrees C. PMID- 16347948 TI - beta-Glucose-1-Phosphate, a Possible Mediator for Polysaccharide Formation in Maltose-Assimilating Lactococcus lactis. AB - Homolactic fermentation of glucose and heterolactic fermentation of maltose with Lactococcus lactis 65.1 were confirmed. When moles of glucose were compared, the uptake rates of the two carbon sources were similar. The intracellular concentration of fructose-1,6-diphosphate (FDP) in maltose-assimilating cells was half of that in glucose-assimilating cells. Similarly, formation of FDP and lactate from maltose by extracts of maltose-grown cells was half of that formed from glucose by extracts of glucose-grown cells, indicating a difference in the utilization of the two carbon sources for energy metabolism. Concentrations of adenine nucleotides were similar in both types of cells. Glucose-1-phosphate was found in extracts of maltose-grown cells given maltose and, in addition, an inducible and low beta-specific phosphoglucomutase activity was observed. beta Glucose-1-phosphate was not metabolized by cell extracts to either FDP or lactate, suggesting an alternative metabolic route. The amount of [C]maltose incorporated into the cell material of maltose-grown cells was four times greater than that of [C]glucose incorporated into the cell material of glucose-grown cells. The intracellular concentration of UTP was lower in maltose-assimilating cells than in glucose-assimilating cells. Cells grown on maltose were more spherical and less fragile than cells grown on glucose. PMID- 16347949 TI - Antibacterial Activity of Sirodesmin PL Phytotoxin: Application to the Selection of Phytotoxin-Deficient Mutants. AB - Sirodesmin PL, a phytotoxin and mycotoxin produced by Leptosphaeria maculans, the causal agent of stem-canker disease of crucifers, exhibited antibacterial activity against gram-positive bacteria and particularly Bacillus subtilis. The importance of the disulfide bridge of the molecule in antibacterial activity was demonstrated. A simple and reliable bioassay based on the antibacterial activity of the toxin was performed for screening sirodesmin PL-deficient mutants when grown on solid culture medium. A mutant was selected and found to produce 3,700 fold less toxin than did the wild-type strain. A sensitive procedure for quantification of the toxin by high-pressure liquid chromatography was developed. Levels of product as low as 100 ng could be detected by this procedure. PMID- 16347950 TI - Effect of yeast hulls on stuck and sluggish wine fermentations: importance of the lipid component. AB - The effect of yeast hulls (yeast ghosts) on sluggish or stuck white wine fermentations was studied. The enhancing effect on yeast growth and fermentation rate displayed by the hulls was shown to be similar to the effect provided by lipid extract from the same hulls. Unsaturated fatty acids and sterols were incorporated into the yeast from lipid extracts during fermentation carried out under oxygen-limited conditions. Adsorption of toxic medium-chain fatty acid (decanoic acid) onto the yeast hulls took place through a dialysis membrane. However, when the hulls were placed inside a dialysis bag, the increase in yeast growth and fermentation rate seen when freely suspended hulls were used did not occur. Accordingly, the effect of yeast hulls in preventing stuck fermentations cannot be attributed only to the adsorption and consequent removal of medium chain fatty acids from the juice. PMID- 16347951 TI - Survival and activity of a 3-chlorobenzoate-catabolic genotype in a natural system. AB - A chlorobenzoate-degrading Alcaligenes strain, BR60, was introduced to flowthrough lake microcosms and exposed to 3-chlorobenzoate (3Cba) concentrations from 0 to 25 muM. A DNA probe specific for BR60 chlorobenzoate catabolic genes was used with the most probable number (MPN) technique to enumerate bacteria harboring this genetic information. This MPN-DNA hybridization method combined with [U-C]3Cba uptake rate measurements allowed the correlation of the size and activity of a specific catabolic population in a natural mixed community for the first time. An experiment involving the release of a streptomycin-resistant strain of BR60 indicated that estimates of bacteria carrying the introduced catabolic genotype often outnumbered plate count estimates of viable BR60 by as much as 3 orders of magnitude, particularly when 3Cba inputs were high. The MPN DNA hybridization method provided catabolic population estimates highly correlated to 3Cba exposure levels and the [U-C]3Cba uptake rates in the microcosms. Plate counts of BR60 were poorly correlated with both 3Cba exposure levels and uptake rates. In the absence of chlorobenzoate selection, the catabolic genotype declined to very low levels by the MPN-DNA hybridization technique after 8 weeks in the microcosms. PMID- 16347952 TI - Competition among Strains of Rhizobium leguminosarum biovar trifolii and Use of a Diallel Analysis in Assessing Competition. AB - Competition between indigenous Rhizobium leguminosarum biovar trifolii strains and inoculant strains or between mixtures of inoculant strains was assessed in field and growth-room studies. Strain effectiveness under competition was compared with strain performance in the absence of competition. Field inoculation trials were conducted at Elora, Ontario, Canada, with soil containing indigenous R. leguminosarum biovar trifolii. The indirect fluorescent-antibody technique was used for the identification of nodule occupants. Treatments consisted of 10 pure strains, a commercial peat inoculant containing a mixture of strains, and an uninoculated control. Inoculant strains occupied 17.5 to 85% of nodules and resulted in increased dry weight and nitrogen content, as compared with the uninoculated control. None of the strains was capable of completely overcoming resident rhizobia, which occupied, on average, 50% of the total nodules tested. In growth-room studies single commercial strains were mixed in all possible two way combinations and assessed in a diallel mating design. Significant differences in plant dry weight of red clover were observed among strain combinations. Specific combining ability effects were significant at the 10% level, suggesting that the effectiveness of strain mixtures depended on the specific strain combinations. Strains possessing superior effectiveness and competitive abilities were identified by field and growth-room studies. No relationship was detected between strain effectiveness and competitive ability or between strain recovery and host cultivar. The concentration of indigenous populations was not considered to be a limiting factor in the recovery of introduced strains at this site. PMID- 16347953 TI - Experimental tests of nutrient limitation in freshwater picoplankton. AB - On the basis of correlative studies, picoplankton in Calder Lake, New York, are apparently unaffected by seasonal fluxes in nutrient (N and P) levels. In this small eutrophic lake, picoplankton (<2.0- to 0.2-mum size) and nanoplankton (<20 to >2 mum) predominate. Microplankton (>20 mum) are typically least important. Experiments were conducted in situ to test whether N, P or N/P ratios affect the predominance of these smaller organisms. Manipulations were run in 4-liter microcosms during June, July, and August 1988, corresponding to periods of increasing stratification and nutrient depletion. Following nutrient additions, phytoplankton were harvested and fractionated into three size classes. Microplankton and nanoplankton were significantly stimulated by both N (2.5 to 50 muM) and P (1 to 20 muM) additions. The severity of nutrient limitation was greatest during July. Picoplankton responded less strongly to N additions and were never P limited. These field data support laboratory studies which indicate that bacterium-sized phytoplankton use nutrients more efficiently and are superior competitors within mixed communities. PMID- 16347954 TI - Pectin Lyase Production by a Penicillium italicum Strain. AB - Growth and concomitant production of an extracellular pectin lyase (PL) [poly(methoxylgalactosiduronate) endolyase; EC 4.2.2.10] were investigated in a group of 16 fungi grown in liquid medium containing pectin as a supplementary carbon source. Culture filtrates of both Penicillium italicum (CECT 2294) and P. expansum (CECT 2275) showed the highest PL activity and contained polygalacturonase but not pectinesterase activity. The effect of the inoculum size, the carbon source (sucrose and glucose syrup), and the presence of pectin on the production of PL by P. italicum was studied. The presence of 2.6 mM glycerophosphate in the culture medium enhanced the appearance of PL but was not inhibitory for the in vitro activity. However, glycerol inhibited the enzyme nearly 50% at such a concentration. PMID- 16347955 TI - Freon 11 extraction of volatile metabolites formed by certain lactic Acid bacteria. AB - The volatile metabolites formed by 18 lactic acid bacteria, representing three genera, were extracted from a complex medium by using a Freon 11 extraction method. The Freon extracts were then analyzed by capillary gas chromatography, and certain extracts were analyzed by gas chromatography-mass spectrometry. A total of 35 major peaks, of which 20 were positively identified, were used to differentiate between the various strains. On the basis of the results obtained, it was possible to differentiate between the members of the genera Lactobacillus, Pediococcus, and Leuconostoc, as well as between various species within the genus Leuconostoc. Of the 10 Leuconostoc oenos strains included in this study, 9 yielded similar results, but it was still possible to differentiate between the various strains. L. oenos B66 differed from the other L. oenos strains. Use of the Freon 11 extraction technique to determine volatile metabolites formed by lactic acid bacteria was shown to be highly reproducible and of great value. Furthermore, certain compounds not previously known to be formed by lactic acid bacteria were found. PMID- 16347956 TI - Novel pathway of toluene catabolism in the trichloroethylene-degrading bacterium g4. AB - o-Cresol and 3-methylcatechol were identified as successive transitory intermediates of toluene catabolism by the trichloroethylene-degrading bacterium G4. The absence of a toluene dihydrodiol intermediate or toluene dioxygenase and toluene dihydrodiol dehydrogenase activities suggested that G4 catabolizes toluene by a unique pathway. Formation of a hybrid species of O- and O-labeled 3 methylcatechol from toluene in an atmosphere of O(2) and O(2) established that G4 catabolizes toluene by successive monooxygenations at the ortho and meta positions. Detection of trace amounts of 4-methylcatechol from toluene catabolism suggested that the initial hydroxylation of toluene was not exclusively at the ortho position. Further catabolism of 3-methylcatechol was found to proceed via catechol-2,3-dioxygenase and hydroxymuconic semialdehyde hydrolase activities. PMID- 16347957 TI - Changes in cytoplasmic carbon and nitrogen pools in a soil bacterium and a fungus in response to salt stress. AB - The effects of water potential on the cellular compositions of a soil bacterium and a fungus were examined by growing the organisms in media with various KCl concentrations. In media containing up to 1 M KCl, C/N ratios in Aspergillus flavus increased significantly, while those for a Pseudomonas sp. did not. For both organisms, the proportions of cellular C and N in cytoplasm increased by a factor of 10 as salinity increased from 0 to 1 M KCl. Such compositional changes have implications for microbial biomass dynamics in soils of varying water potential and for biomass measurement by chloroform fumigation. PMID- 16347958 TI - Thermostable, Raw-Starch-Digesting Amylase from Bacillus stearothermophilus. AB - An endospore-forming thermophilic bacterium, which produced amylase and was identified as Bacillus stearothermophilus, was isolated from soil. The amylase had an optimum temperature of 70 degrees C and strongly degraded wheat starch granules (93%) and potato starch granules (80%) at 60 degrees C. PMID- 16347959 TI - Degradation of lignin-related compounds by actinomycetes. AB - Evidence for activity against the lignin fraction of straw was produced for a range of actinomycete strains. Decolorization of the polymeric dye Poly R and oxidation of veratryl alcohol, indicators of ligninolytic activity in white rot fungi, and utilization of fractionated Kraft lignin and low-molecular-weight methoxylated aromatic compounds were the criteria used. The relationships between these activities and the solubilization of native lignin are discussed. PMID- 16347960 TI - Threshold acetate concentrations for acetate catabolism by aceticlastic methanogenic bacteria. AB - [This corrects the article on p. 514 in vol. 55.]. PMID- 16347961 TI - Transformation of Tetrachloromethane to Dichloromethane and Carbon Dioxide by Acetobacterium woodii. AB - [This corrects the article on p. 2822 in vol. 54.]. PMID- 16347962 TI - Isolation and Characterization of an Enterobacter cloacae Strain That Reduces Hexavalent Chromium under Anaerobic Conditions. AB - An Enterobacter cloacae strain (HO1) capable of reducing hexavalent chromium (chromate) was isolated from activated sludge. This bacterium was resistant to chromate under both aerobic and anaerobic conditions. Only the anaerobic culture of the E. cloacae isolate showed chromate reduction. In the anaerobic culture, yellow turned white with chromate and the turbidity increased as the reduction proceeded, suggesting that insoluble chromium hydroxide was formed. E. cloacae is likely to utilize toxic chromate as an electron acceptor anaerobically because (i) the anaerobic growth of E. cloacae HO1 accompanied the decrease of toxic chromate in culture medium, (ii) the chromate-reducing activity was rapidly inhibited by oxygen, and (iii) the reduction occurred more rapidly in glycerol- or acetate-grown cells than in glucose-grown cells. The chromate reduction in E. cloacae HO1 was observed at pH 6.0 to 8.5 (optimum pH, 7.0) and at 10 to 40 degrees C (optimum, 30 degrees C). PMID- 16347963 TI - Survival of Ice Nucleation-Active and Genetically Engineered Non-Ice-Nucleating Pseudomonas syringae Strains after Freezing. AB - The survival after freezing of ice nucleation-active (INA) and genetically engineered non-INA strains of Pseudomonas syringae was compared. Each strain was applied to oat seedlings and allowed to colonize for 3 days, and the plants were subjected to various freezing temperatures. Plant leaves were harvested before and after freezing on two consecutive days, and bacterial populations were determined. Populations of the INA wild-type strain increased 15-fold in the 18 h after the oat plants incurred frost damage at -5 and -12 degrees C. Plants colonized by the non-INA strain were undamaged at -5 degrees C and exhibited no changes in population size after two freeze trials. As freezing temperatures were lowered (-7, -9, and -12 degrees C), oat plants colonized by the non-INA strain suffered increased frost damage concomitant with bacterial population increases following 18 h. At -12 degrees C, both strains behaved identically. The data show a relationship between frost damage to plants and increased bacterial population size during the following 18 h, indicating a potential competitive advantage of INA strains of P. syringae over non-INA strains in mild freezing environments. PMID- 16347964 TI - Influence of Bradyrhizobium japonicum Location and Movement on Nodulation and Nitrogen Fixation in Soybeans. AB - The influence of seed and soil inoculation on bradyrhizobial migration, nodulation, and N(2) fixation was examined by using two Bradyrhizobium japonicum strains of contrasting effectiveness in N(2) fixation. Seed-inoculated strains formed fewer nodules on soybeans (mostly restricted to the tap and crown roots within 0 to 5 cm from the stem base) than did bradyrhizobia distributed throughout the soil or inoculated at specific depths. Nodulation was greater below the depths at which bradyrhizobial cells were located rather than above, even though watering was done from below to minimize passive bradyrhizobial migration with percolating water. The most profuse nodulation occurred within approximately 5 cm below the point of placement and was generally negligible below 10 cm. These and other results suggest that bradyrhizobial migration from the initial point of placement was very limited. Nevertheless, the more competitive strain, effective strain THA 7, migrated into soil to a greater extent than the ineffective strain THA 1 did. Nitrogen fixation resulting from the dual-strain inoculations differed depending on the method of inoculation. For example, the amount of N(2) fixed when both strains were slurried together onto the seed was about half that obtained from mixing the effective strain into the soil with the ineffective strain on the seed. The results indicate the importance of rhizobial distribution or movement into soil for nodulation, nodule distribution, strain competitiveness, and N(2) fixation in soil-grown legumes. PMID- 16347965 TI - Isolation and Characterization of Aminopeptidase-Deficient Lactobacillus bulgaricus Mutants. AB - Lactobacillus bulgaricus CNRZ 397 is able to hydrolyze many amino-acyl- and dipeptidyl-beta-naphthylamides. Analysis of heat inactivation kinetics, protease inhibitor effects, and the subcellular location of aminopeptidase (AP) activities from the parental strain and mutant derivatives dificient in alanyl- or leucyl beta-naphthylamide hydrolysis pointed out the existence of four APs. All mutants isolated were totally deficient in AP II, a cell wall metallo-enzyme with a broad substrate specificity but that is specifically responsible for lysyl-AP activity and is characterized by a molecular mass of 95,000 daltons. AP I and AP III are cytoplasmic enzymes that exhibit arginyl-AP activity; both enzymes are inducible during growth in rich peptide MRS medium (Difco Laboratories, Detroit, Mich.). The existence of a fourth AP (AP IV) that is involved in leucyl-AP activity was suggested. Moreover, we showed that X-prolyl-dipeptidyl-AP activity, which was not catalyzed by an AP, involved an enzyme(s) that is controlled by a regulatory mechanism that is common to that of AP II. PMID- 16347967 TI - Genetic Analysis of Haploids from Industrial Strains of Baker's Yeast. AB - Strains of baker's yeast conventionally used by the baking industry in Japan were tested for the ability to sporulate and produce viable haploid spores. Three isolates which possessed the properties of baker's yeasts were obtained from single spores. Each strain was a haploid, and one of these strains, YOY34, was characterized. YOY34 fermented maltose and sucrose, but did not utilize galactose, unlike its parental strain. Genetic analysis showed that YOY34 carried two MAL genes, one functional and one cryptic; two SUC genes; and one defective gal gene. The genotype of YOY34 was identified as MATalpha MAL1 MAL3g SUC2 SUC4 gall. The MAL1 gene from this haploid was constitutively expressed, was dominant over other wild-type MAL tester genes, and gave a weak sucrose fermentation. YOY34 was suitable for both bakery products, like conventional baker's yeasts, and for genetic analysis, like laboratory strains. PMID- 16347966 TI - Diffusion of the Interspecies Electron Carriers H(2) and Formate in Methanogenic Ecosystems and Its Implications in the Measurement of K(m) for H(2) or Formate Uptake. AB - We calculated the potential H(2) and formate diffusion between microbes and found that at H(2) concentrations commonly found in nature, H(2) could not diffuse rapidly enough to dispersed methanogenic cells to account for the rate of methane synthesis but formate could. Our calculations were based on individual organisms dispersed in the medium, as supported by microscopic observations of butyrate degrading cocultures. We isolated an axenic culture of Syntrophomonas wolfei and cultivated it on butyrate in syntrophic coculture with Methanobacterium formicicum; during growth the H(2) concentration was 63 nM (10.6 Pa). S. wolfei contained formate dehydrogenase activity (as does M. formicicum), which would allow interspecies formate transfer in that coculture. Thus, interspecies formate transfer may be the predominant mechanism of syntrophy. Our diffusion calculations also indicated that H(2) concentration at the cell surface of H(2) consuming organisms was low but increased to approximately the bulk-fluid concentration at a distance of about 10 mum from the surface. Thus, routine estimation of kinetic parameters would greatly overestimate the K(m) for H(2) or formate. PMID- 16347968 TI - N(2) fixation by bacteria associated with maize roots at a low partial o(2) pressure. AB - Nitrogen fixation by bacteria associated with roots of intact maize plants was measured by exposing the roots to N(2) at a partial O(2) pressure (pO(2)) of 2 or 10 kPa. The plants were grown in a mixture of Weswood soil and sand and then transferred to plastic cylinders containing an N-free plant nutrient solution. The solution was sparged continuously with a mixture of air and N(2) at a pO(2) of 2 or 10 kPa. Acetylene reduction was measured after the roots were exposed to the low pO(2) overnight. The air-N(2) atmosphere in the cylinders was then replaced with an O(2)-He atmosphere at the same pO(2), and the roots were exposed to 20 kPa of N(2) for 20 to 22 h. Incorporation of N into the roots was 200 times greater at 2 kPa of O(2) than at 10 kPa of O(2). Adding l-malate (1 g of C liter) to the nutrient solution increased root-associated nitrogenase activity, producing a strong N label which could be traced into the shoots. Fixed N was detected in the shoots within 5 days after the plants were returned to unfertilized soil. In a similar experiment with undisturbed plants grown in fritted clay, movement of fixed N into the shoots was evident within 4 days after the roots were exposed to N(2) at 2 kPa of O(2). Inoculation with Azospirillum lipoferum yielded no significant differences in shoot dry weight, total nitrogen content, percent nitrogen, or N enrichment of plant tissues. Inoculated plants did exhibit greater root dry weight than uninoculated plants, however. PMID- 16347969 TI - Effect of Na Concentration and Nutritional Factors on the Lag Phase and Exponential Growth Rates of the Marine Bacterium Deleya aesta and of Other Marine Species. AB - Growth of the marine bacterium Deleya aesta in a succinate minimal medium showed increasingly long lag phases as Na was decreased below the optimum (200 to 500 mM). The minimum Na concentration permitting growth consistently was 15 mM. Supplementation of the medium with KHCO(3) (as a source of CO(2)) or yeast extract, especially in combination, reduced the lag phase, increased the rate of exponential growth, and allowed growth at 8 mM Na. KHCO(3) did not reduce the lag period but did increase the rate of exponential growth of Deleya venusta, Deleya pacifica, and Alteromonas haloplanktis 214. Yeast extract was active for all three. The effect of yeast extract on D. aesta could be reproduced by a mixture of amino acids approximating its amino acid composition. l-Alanine, l-aspartate, and l-methionine, in combination, were the most effective in reducing the lag phase, although not as effective as the complete mixture. Succinate, l-aspartate, and l-alanine were transported into the cells by largely independent pathways and oxidized at rates which were much lower at 10 than at 200 mM Na. l-Methionine was transported at a low rate in the absence of Na and at a higher rate at 10 mM but was not oxidized. Above 25 mM Na, the rate of transport of the carbon source was not the rate-limiting step for growth. It is concluded that a combination of transportable carbon sources reduced the lag period and increased the rate of exponential growth because they can be taken up independently and at low Na utilized simultaneously. PMID- 16347970 TI - Effects of fixation on cell volume of marine planktonic protozoa. AB - The effects of fixation on the cell volume of marine heterotrophic nanoflagellates and planktonic ciliates were investigated. Decreases in cell volume depended on the combination of the protozoan taxa and the particular fixative. For a particular fixative and protozoan species, degree of shrinkage was independent of physiological state. The volume of fixed cells was found to be approximately 20 to 55% lower than the cell volume of live organisms. For the heterotrophic microflagellates, the fixatives ranked, in order of decreasing effect on cell volume, as glutaraldehyde, formaldehyde, acid Lugol's solution, and modified van der Veer solution. With oligotrichous ciliates and a tintinnid ciliate, formaldehyde caused less shrinkage than glutaraldehyde or acid Lugol's solution. With the aldehyde fixatives, the microflagellates were found to shrink more than the ciliates. Differential effects of fixation on cell volumes may result in an underestimation of the biomass of certain protozoan taxa in natural samples. PMID- 16347971 TI - Comparison of two methods for enumeration of anaerobe numbers on forages and evaluation of ethylene oxide treatment for forage sterilization. AB - Experiments were conducted to (i) compare most-probable-number (MPN) procedures with roll tube procedures for enumeration of forage anaerobic bacteria and (ii) evaluate the efficacy of using ethylene oxide to sterilize wet herbage. Alfalfa, corn, and alfalfa-orchardgrass silages and alfalfa and orchardgrass herbages were analyzed for total anaerobic bacteria (medium pH, 6.8) and acid-tolerant anaerobic bacteria (medium pH, 4.5) by both roll tube and MPN procedures. No difference was found between the roll tube and MPN procedures for total bacteria; however, higher counts were obtained for acid-tolerant bacteria when the MPN procedure was used. Although MPN procedures require less time to obtain an estimate of bacterial numbers, isolation and identification of the microbial population is not possible. Alfalfa herbage was treated with ethylene oxide for 12, 24, or 36 h, incubated for 7 days at 37 degrees C with or without addition of a bacterial inoculant, and analyzed for total bacteria by MPN procedures. Microbial growth after inoculation of ethylene oxide-treated herbage indicated that there was insufficient residual ethylene oxide to inhibit subsequent microbial growth. The results also indicated that 24 h was required to adequately sterilize fresh herbage. Thus, ethylene oxide can be used to sterilize wet herbage for use as a substrate for pure cultures of silage bacteria. PMID- 16347972 TI - Protozoan grazing and bacterial production in stratified lake vechten estimated with fluorescently labeled bacteria and by thymidine incorporation. AB - In stratified Lake Vechten, The Netherlands, protozoan grazing was estimated on the basis of uptake of fluorescently labeled bacteria and compared with bacterial production estimated on the basis of thymidine incorporation. By using a grazer free mixed bacterial population from the lake in continuous culture, an empirical relationship between cell production and thymidine incorporation was established. Thymidine incorporation into total cold-trichloroacetic-acid-insoluble macromolecules yielded a relatively constant empirical conversion factor of ca. 10 (range, 0.38 x 10 to 1.42 x 10) bacteria mol of thymidine at specific growth rates (mu) ranging from 0.007 to 0.116 h. Although thymidine incorporation has been assumed to measure DNA synthesis thymidine incorporation appeared to underestimate the independently measured bacterial DNA synthesis by at least 1.5- to 13-fold, even if all incorporated label was assumed to be in DNA. However, incorporation into DNA was found to be insignificant as measured by conventional acid-base hydrolysis. Methodological problems of the thymidine technique are discussed. Like the cultures, Lake Vechten bacteria showed considerable thymidine incorporation into total macromolecules, but no significant incorporation into DNA was found by acid-base hydrolysis. This applied not only to the low-oxygen hypo- and metalimnion but also to the aerobic epilimnion. Thus, the established empirical conversion factor for thymidine incorporation into total macromolecules was used to estimate bacterial production. Maximum production rates (141 x 10 bacteria liter h; mu, 0.012 h) were found in the metalimnion and were 1 order of magnitude higher than in the epi- and hypolimnion. In all three strata, the estimated bacterial production was roughly balanced by the estimated protozoan grazing. Heterotrophic nanoflagellates were the major consumers of the bacterial production and showed maximum numbers (up to 40 x 10 heterotrophic nanoflagellates liter) in the microaerobic metalimnion. PMID- 16347973 TI - Determination of biomass by ultrasonic measurements. AB - The quantitative determination of biomass in a suspension by means of ultrasound velocity is a simple and on-line-applicable method. Such an ultrasonic sensor offers the advantage of being long-term stable, reliable, and sterilizable. In this paper we present sound velocity measurements made with different microorganisms. The experimental results which we have obtained with an impulse echo method will be compared with theoretical predictions and discussed with respect to previous findings (Y. Ishimori, I. Karube, and S. Suzuki, Appl. Environ. Microbiol. 42:632-637, 1981). PMID- 16347974 TI - Catechol Formation and Melanization by Na -Dependent Azotobacter chroococcum: a Protective Mechanism for Aeroadaptation? AB - Aeroadaptive microaerophilic Azotobacter chroococcum 184 produced a cell associated black pigment when grown at high aeration rates under nitrogen-fixing conditions. This pigment was shown to be a catechol melanin. Polyphenol oxidase activity was detected in cell extracts of cells grown for 72 h. Melanin formation was optimal in the later stages of growth, and there was no correlation between nitrogenase activity and melanization. Nitrogenase activity in strain 184 was optimal at 10% O(2), and melanin formation was suppressed by O(2) limitation. In the presence of charcoal, an adsorbent of toxic oxygen intermediates, and benzoic acid, a scavenger of hydroxyl radicals, melanization was inhibited. However, in the presence of copper, the intensity of pigment color increased and melanization was accelerated. Copper also eliminated catalase and peroxidase activities of the organism but still permitted aerobic growth. In the presence of low levels of iron, melanization was accelerated under high aeration rates, and under low rates of aeration, melanization was observed only at higher levels of iron. Hydroxamate siderophore production was detectable in the presence of soluble iron under high rates of aeration but was repressed by the same levels of iron under low aeration rates. Unlike melanization and hydroxamate formation, catechol formation was observed under both low and high rates of aeration under nitrogen-fixing conditions. Catechol formation and melanization were repressed by 14 mM NH(4), at which level nitrogenase activity was also repressed. Copper reversed the repressive effect of NH(4). A role for catechol formation and melanization in aeroadaptation is proposed. PMID- 16347975 TI - Selective Recovery of 16S rRNA Sequences from Natural Microbial Communities in the Form of cDNA. AB - Cloning of cDNA obtained from 16S rRNA (16S rcDNA) selectively retrieves species specific sequence information useful for analyzing the composition and structure of natural microbial communities. With this technique we obtained recombinant 16S rcDNA libraries from Escherichia coli and from a model hot-spring cyanobacterial mat community. The recombinant plasmids contained exclusively 16S rRNA-derived inserts. This selective approach is independent of biasing culture techniques and eliminates the laborious screening required to locate 16S rRNA gene-bearing recombinants in genomic DNA libraries obtained from natural communities. PMID- 16347976 TI - Turnover of extracellular DNA in eutrophic and oligotrophic freshwater environments of southwest Florida. AB - The turnover of extracellular DNA was investigated in oligotrophic springs of the Crystal River and the eutrophic Medard Reservoir of southwest Florida. The Medard Reservoir possessed large populations of bacterioplankton and phytoplankton (6.8 x 10 cells per liter and 28.6 mug of chlorophyll a per liter, respectively), while the Crystal River springs only contained a fraction of the microbial biomass found in the Medard Reservoir. Although dissolved DNA values were greater in the Medard Reservoir, higher rates of DNA removal resulted in similar extracellular DNA turnover times in both environments (9.62 +/- 3.6 h in the Crystal River and 10.5 +/- 2.1 h in the Medard Reservoir). These results indicate that regardless of trophic status or microbial standing stock, extracellular DNA turns over rapidly in subtropical planktonic freshwater environments. Therefore, recombinant DNA sequences from released genetically engineered microorganisms might not be expected to survive for long periods of time in freshwater planktonic environments. PMID- 16347977 TI - Molybdenum Involvement in Aerobic Degradation of 2-Furoic Acid by Pseudomonas putida Fu1. AB - An organism identified as Pseudomonas putida was isolated from an enrichment culture with 2-furoic acid as its sole source of carbon and energy. The organism contained a 2-furoyl-coenzyme A (CoA) synthetase to form 2-furoyl-CoA and a 2 furoyl-CoA dehydrogenase to form 5-hydroxy-2-furoyl-CoA as the first two enzymes involved in the degradation. Tungstate, the specific antagonist of molybdate, decreased growth rate and consumption of 2-furoic acid but had no influence on growth with succinate. Correspondingly, the 2-furoyl-CoA dehydrogenase activity decreased when the organism was grown on 2-furoic acid in the presence of increasing amounts of tungstate. The addition of molybdate reversed the negative effect on 2-furoyl-CoA dehydrogenase activity, which points to the involvement of a molybdoenzyme in this reaction. Both enzymes studied were inducible. No plasmid was detected in this organism. PMID- 16347978 TI - Production and Utilization of Ethanol by the Homoacetogen Acetobacterium woodii. AB - Acetobacterium woodii formed ethanol as a fermentation product in addition to acetate when the phosphate concentration of the medium was between 0.2 and 8.4 mM. Considerable amounts of alanine were also found (2 to 11 mM). Supplementation with phosphate caused a shift to acetate as the only end product. Ethanol could also serve as a substrate for A. woodii. The fermentation yielded predominantly acetate and was strictly dependent on high bicarbonate concentrations. 1 Propanol, 1-butanol, and 1-pentanol were converted to the corresponding fatty acids but allowed only marginal growth. A. wieringae and A. carbinolicum grown under identical conditions were also able to form ethanol, and A. wieringae could use ethanol as a substrate, too. Alcohol dehydrogenase and acetaldehyde dehydrogenase activities were determined in A. woodii. Activity stains of polyacrylamide gels with crude extracts allowed the detection of acetaldehyde dehydrogenase but not of alcohol dehydrogenase. Trace amounts of methane were detected during growth of A. woodii on glucose and ethanol. PMID- 16347979 TI - Annual pattern of denitrification and nitrate ammonification in estuarine sediment. AB - The seasonal variation and depth distribution of the capacity for denitrification and dissimilatory NO(3) reduction to NH(4) (NO(3) ammonification) were studied in the upper 4 cm of the sediment of Norsminde Fjord estuary, Denmark. A combination of C(2)H(2) inhibition and N isotope techniques was used in intact sediment cores in short-term incubations (maximum, 4 h). The denitrification capacity exhibited two maxima, one in the spring and one in the fall, whereas the capacity for NO(3) ammonification was maximal in the late summer, when sediments were progressively reduced. The denitrification capacity was always highest in the uppermost 1 cm of the sediment and declined with depth. The NO(3) ammonification was usually higher with depth, but the maximum activity in late summer was observed within the upper 1 cm. The capacity for NO(3) incorporation into organic material was investigated on two occasions in intact sediment cores and accounted for less than 5% of the total NO(3) reduction. Denitrification accounted for between 13 and 51% of the total NO(3) reduction, and NH(4) production accounted for between 4 and 21%, depending on initial rates during the time courses. Changes of the rates during the incubation were observed in the late summer, which reflected synthesis of denitrifying enzymes. This time lag was eliminated in experiments with mixed sediment because of preincubation with NO(3) and alterations of the near environmental conditions. The initial rates obtained in intact sediment cores therefore reflect the preexisting enzyme content of the sediment. PMID- 16347980 TI - Exopolysaccharide-Deficient Mutants of Rhizobium fredii HH303 Which Are Symbiotically Effective. AB - Nineteen Tn5-induced mutants of Rhizobium fredii HH303 defective in acidic exopolysaccharide synthesis were isolated by screening for lack of Calcofluor fluorescence. They were grouped by complementation analysis by using Rhizobium meliloti cosmids carrying exo genes. All of the 19 mutants were symbiotically effective or partially effective, indicating that the major bacterial acidic exopolysaccharide of this strain of R. fredii may not be required for symbiotic development in the soybean. PMID- 16347981 TI - Improvement of Rhizobium inoculants. AB - [This corrects the article on p. 865 in vol. 55.]. PMID- 16347982 TI - Combined Effect of Water Activity and pH on Inhibition of Toxin Production by Clostridium botulinum in Cooked, Vacuum-Packed Potatoes. AB - [This corrects the article on p. 658 in vol. 55.]. PMID- 16347983 TI - Protozoan Response to the Addition of Bacterial Predators and Other Bacteria to Soil. AB - Representatives of several categories of bacteria were added to soil to determine which of them might elicit responses from the soil protozoa. The various categories were nonobligate bacterial predators of bacteria, prey bacteria for these predators, indigenous bacteria that are normally present in high numbers in soil, and non-native bacteria that often find their way in large numbers into soil. The soil was incubated and the responses of the indigenous protozoa were determined by most-probable-number estimations of total numbers of protozoa. Although each soil was incubated with only one species of added bacteria, the protozoan response for the soil was evaluated by using most-probable-number estimations of several species of bacteria. The protozoa did not respond to incubation of the soil with either Cupriavidus necator, a potent bacterial predator, or one of its prey species, Micrococcus luteus. C. necator also had no effect on the protozoa. Therefore, in this case, bacterial and protozoan predators did not interact, except for possible competition for bacterial prey cells. The soil protozoa did not respond to the addition of Arthrobacter globiformis or Bacillus thuringiensis. Therefore, the autochthonous state of Arthrobacter species in soil and the survival of B. thuringiensis were possibly enhanced by the resistance of these species to protozoa. The addition of Bacillus mycoides and Escherichia coli cells caused specific responses by soil protozoa. The protozoa that responded to E. coli did not respond to B. mycoides or any other bacteria, and vice versa. Therefore, addition to soil of a nonsoil bacterium, such as E. coli, did not cause a general increase in numbers of protozoa or in protozoan control of the activities of other bacteria in the soil. PMID- 16347984 TI - Maintenance of Intracellular pH and Acid Tolerance in Rhizobium meliloti. AB - The development and function of the Rhizobium meliloti-Medicago sp. symbiosis are sensitive to soil acidity. Physiological criteria that can be measured in culture which serve to predict acid tolerance in soil would be valuable. The intracellular pH of R. meliloti was measured using either radioactively labeled weak acids (5,5-dimethyloxazolidine-2,4-dione and butyric acid) or pH-sensitive fluorescent compounds; both methods gave similar values. Six acid-tolerant strains (WSM419, WSM533, WSM539, WSM540, WSM852, and WSM870) maintained an alkaline intracellular pH when the external pH was between 5.6 and 7.2. In contrast, two Australian commercial inoculant strains (CC169 and U45) and four acid-sensitive strains from alkaline soils in Iraq (WSM244, WSM301, WSM365, and WSM367) maintained an alkaline intracellular pH when the external pH was >/=6.5, but had intracellular pH values of Cd > Co > Zn > Ni > Cu > Cr > Mn. PMID- 16347996 TI - Bacteriophage Transport in Sandy Soil and Fractured Tuff. AB - Bacteriophage transport was investigated in laboratory column experiments using sandy soil, a controlled field study in a sandy wash, and laboratory experiments using fractured rock. In the soil columns, the phage MS-2 exhibited significant dispersion and was excluded from 35 to 40% of the void volume but did not adsorb. Dispersion in the field was similiar to that observed in the laboratory. The phage f2 was largely excluded from the porous matrix of the two fractured-rock cores studied, coming through 1.2 and 2.0 times later than predicted on the basis of fracture flow alone. Because of matrix diffusion, nonsorbing solutes were retarded by over a factor of three relative to fracture flow. The time for a solute tracer to equilibrate with the porous matrix of 6.5-cm-diameter by 25-cm long cores was measured in days. Results of both granular-medium and fractured rock experiments illustrate the inability of a solute tracer to provide estimates for dispersion and effective porosity that are applicable to a colloid. Bacteriophage can be used to better estimate the maximum subsurface transport rate of colloidal contaminants through a porous formation. PMID- 16347997 TI - Induction of Lipomyces starkeyi Dextranase. AB - Lipomyces starkeyi ATCC 20825 is a derepressed mutant derived from L. starkeyi ATCC 12659. It requires the presence of an inducer before it produces dextranase. This study was undertaken to determine the most efficient, commercially feasible method for inducing this enzyme. The following compounds induced dextranase synthesis: 1-O-beta-methyl-glucopyranoside, 1-O-alpha-methyl-glucopyranoside, dextran, isomaltopentose, isomaltotetraose, isomaltotriose, and isomaltose. 1-O beta-Methyl-glucopyranoside was found to be a gratuitous inducer. Early in the growth phase, cells produced higher specific levels of enzyme than they did in late log phase. The length of exposure of the yeast cells to the inducer also affected the amount of dextranase produced. The maximum amount of enzyme was produced after 12 h of exposure to the inducer. The saturation concentration was the same for all inducers tested, i.e., approximately 1 mg of inducer for every 2 x 10 cells. PMID- 16347995 TI - Localization of Bacteria and Hemoglobin in Root Nodules of Parasponia andersonii Containing Both Bradyrhizobium Strains and Rhizobium leguminosarum biovar trifolii. AB - Dual occupancy of Parasponia andersonii nodules with different Bradyrhizobium strains and Rhizobium leguminosarum biovar trifolii was frequently obtained when two strains were inoculated into plants grown aseptically in tubes. Since reisolates of Bradyrhizobium strains from dually occupied nodules acquired the ability to nodulate Trifolium repens, the spatial relationship of the two species of bacteria during nodule initiation and development was investigated and their proximity was demonstrated. By using light microscopy and electron microscopy and immunogold labeling, R. leguminosarum biovar trifolii NGR66 inoculated alone onto P. andersonii produced small ineffective nodules, with bacteria embedded in matrix material in intercellular spaces and in a few nonliving host cells rather than in infection threads (CP299). In dual infections, the two bacterial species were shown to be adjacent to one another in the matrix of nodule intercellular spaces and in some host nodule cells. However, when two different Bradyrhizobium strains occupied a single nodule, they were located in different lobes of the same nodule. Immunogold labeling showed that Parasponia hemoglobin was localized in the cytoplasm of young infected nodule cells. This suggests that the nitrogen fixing phase of Parasponia nodule cells is short-lived and correlates with previous acetylene reduction data from nodule slices. Hemoglobin was associated only with areas of nodule tissue infected with the effective nitrogen-fixing strain CP299 and absent from areas infected with R. leguminosarum biovar trifolii. PMID- 16347998 TI - Cultivation Techniques for Hyperthermophilic Archaebacteria: Continuous Culture of Pyrococcus furiosus at Temperatures near 100 degrees C. AB - A system which allows continuous cultivation of hyperthermophilic archaebacteria at temperatures approaching 100 degrees C has been developed. Continuous cultivation of the hyperthermophile Pyrococcus furiosus was carried out with this system; the resulting dilution rate and gas production profiles are discussed. PMID- 16347999 TI - Mineralization of Surfactants by Microbiota of Aquatic Plants. AB - The biodegradation of linear alkylbenzene sulfonate (LAS) and linear alcohol ethoxylate (LAE) by the microbiota associated with duckweed (Lemna minor) and the roots of cattail (Typha latifolia) was investigated. Plants were obtained from a pristine pond and a pond receiving wastewater from a rural laundromat. Cattail roots and duckweed plants were incubated in vessels containing sterile water amended with [C]LAS, [C]LAE, or C-labeled mixed amino acids (MAA). Evolution of CO(2) was determined over time. The microbiota of cattail roots from both ponds mineralized LAS, LAE, and MAA without lag periods, and the rates and extents of mineralization were not significantly affected by the source of the plants. Mineralization of LAS and LAE was more rapid in the rhizosphere than in nearby root-free sediments, which exhibited differences as a function of pond. The microbiota of duckweed readily mineralized LAE and MAA but not LAS. The rate and extent of mineralization were not affected by the source of the duckweed. PMID- 16348000 TI - Cadmium Resistance Screening in Nitrilotriacetate-Buffered Minimal Media. AB - Media used to determine the MICs of heavy metals for bacteria are unreliable because organic components in the media bind or chelate most of the metal being studied. To define specific metal activity in media and to maintain metal activity at a constant level, HEPES-MES [N-2-hydroxyethylpiperazine-N' -2 ethanesulfonic acid-2-(N-morpholine)ethanesulfonic acid] salts medium with arabinose medium was modified, and the modified medium was used to examine the MIC of cadmium for Rhizobium fredii USDA 201. Arabinose-HEPES-MES was modified by addition of the chelator nitrilotriacetate to buffer the supply of free Cd ion to maintain a constant Cd activity and by the use of only MES to buffer pH (buffered arabinose-MES medium [BAM]). Ca and Mg were supplied at the normal levels for soil solutions, and other trace elements were supplied at the levels required for normal growth of plants. The concentration of free Cd ion was calculated by using the computer program GEOCHEM-PC with a corrected data base. The Cd MIC in BAM was 14.0 muM, while that in a tryptone-yeast extract medium was 107 muM. The results indicate that substantial free Cd is removed from solution in most standard media, resulting in falsely high MICs. The new BAM medium allows for the precise determination of MICs, thus avoiding the uncertainties associated with other media. PMID- 16348001 TI - The K1 Toxin of Saccharomyces cerevisiae Kills Spheroplasts of Many Yeast Species. AB - The Saccharomyces cerevisiae K1 toxin killed spheroplasts from the genera Candida, Kluyveromyces, and Schwanniomyces. Cells of these organisms were toxin insensitive. The toxin bound poorly to Kluyveromyces lactis cells. In contrast, Candida albicans bound the toxin to an extent similar to that seen with S. cerevisiae. Thus, wall receptors can define toxin specificity and are necessary but not sufficient for toxin action on intact cells. PMID- 16348002 TI - Nitrosovibrio spp., the Dominant Ammonia-Oxidizing Bacteria in Building Sandstone. AB - In five historical buildings in the Federal Republic of Germany, ammonia oxidizing bacteria of the genera Nitrosovibrio, Nitrosospira, and Nitrosomonas were detected in high cell numbers. In building stones, Nitrosovibrio was the most abundant ammonia-oxidizing organism. In the soil at the foot of each building, Nitrosomonas spp. were the most common ammonia oxidizers, whereas Nitrosovibrio spp. were not detected. PMID- 16348003 TI - Production of Major Extracellular Enzymes during Lignocellulose Degradation by Two Streptomycetes in Agitated Submerged Culture. AB - [This corrects the article on p. 1167 in vol. 55.]. PMID- 16348004 TI - Continuous-Culture Responses of Candida shehatae to Shifts in Temperature and Aeration: Implications for Ethanol Inhibition. AB - Temperature and aeration shifts were used to perturb steady-state continuous cultures to determine the effects of ethanol on xylose metabolism by Candida shehatae. The accumulation of ethanol exerted a delayed inhibitory effect on the specific rate of substrate utilization. A second effect was also observed in which the specific rate of xylitol production increased at the expense of the specific rate of ethanol production. Both effects were enhanced at higher temperature. Inhibitory effects also occurred in glucose metabolism. PMID- 16348005 TI - Synergism in degradation and utilization of intact forage cellulose, hemicellulose, and pectin by three pure cultures of ruminal bacteria. AB - Pure cultures of ruminal bacteria characterized as using only a single forage polysaccharide (Fibrobacter succinogenes A3c, cellulolytic; Bacteroides ruminicola H2b, hemicellulolytic; Lachnospira multiparus D15d, pectinolytic) were inoculated separately and in all possible combinations into fermentation tubes containing orchard grass as the sole substrate. Fermentations were run to completion, and then cultures were analyzed for digestion of cellulose plus degradation and utilization of hemicellulose and pectin. Addition of the noncellulolytic organisms, in any combination, to the cellulolytic organism F. succinogenes had little effect on overall cellulose utilization. F. succinogenes degraded but could not utilize hemicellulose; however, when it was combined with B. ruminicola, total utilization of hemicellulose increased markedly over that by B. ruminicola alone. L. multiparus was inactive in hemicellulose digestion, alone or in any combination. Although unable to degrade and utilize purified pectin, B. ruminicola degraded and utilized considerable quantities of the forage pectin. In contrast, L. multiparus was very active against purified pectin, but had extremely limited ability to degrade and utilize pectin from the intact forage. Both degradation and utilization of forage pectin increased when F. succinogenes was combined with B. ruminicola. Sequential addition of two cultures, allowing one to complete its fermentation before adding the second, was used to study synergism between cultures on forage pectin digestion. In general, synergistic effects did not appear to be related to a particular sequence of utilization. The ability of F. succinogenes to degrade and B. ruminicola to degrade and utilize forage pectin contradicts both previous and present data obtained with purified pectin. Thus, isolation and characterization of ruminal bacteria on purified substrates may be misleading with regard to their role in the overall ruminal fermentation. PMID- 16348006 TI - Acetate production by methanogenic bacteria. AB - Methanosarcina barkeri MS and 227 and Methanosarcina mazei S-6 produced acetate when grown on H(2)-CO(2), methanol, or trimethylamine. Marked differences in acetate production by the two bacterial species were found, even though methane and cell yields were nearly the same. M. barkeri produced 30 to 75 mumol of acetate per mmol of CH(4) formed, but M. mazei produced only 8 to 9 mumol of acetate per mmol of CH(4). PMID- 16348007 TI - Modification of Lignins by Growing Cells of the Sulfate-Reducing Anaerobe Desulfovibrio desulfuricans. AB - The anaerobic sulfate-reducing bacterium Desulfovibrio desulfuricans was grown on medium supplemented with either Kraft lignin or lignosulfonate. Only lignosulfonate contributed to the growth of D. desulfuricans cells, by replacing sulfate, a natural electron acceptor for this microorganism. Kraft lignin added to the culture medium could not substitute for lactate or sulfate, both necessary culture medium components. However, it was found to enhance the viability of D. desulfuricans cells. When changes occurring in lignin during growth of Desulfovibrio cultures were monitored, it was found that both lignin preparations could be partially depolymerized. Spectrophotometric and elemental analysis of biologically treated lignins suggested that both the polyphenolic backbone and lignin functional groups were affected by D. desulfuricans. After treatment, a twofold increase in the sulfur content of Kraft lignin and a minor decrease (14%) in the sulfur content of lignosulfonate were observed. After biological treatment, Kraft lignin and lignosulfonate both bound larger quantities of heavy metals. PMID- 16348008 TI - Degradation of Acetonitrile by Pseudomonas putida. AB - A bacterium capable of utilizing high concentrations of acetonitrile as the sole source of carbon and nitrogen was isolated from soil and identified as Pseudomonas putida. This bacterium could also utilize butyronitrile, glutaronitrile, isobutyronitrile, methacrylonitrile, propionitrile, succinonitrile, valeronitrile, and some of their corresponding amides, such as acetamide, butyramide, isobutyramide, methacrylamide, propionamide, and succinamide as growth substrates. Acetonitrile-grown cells oxidized acetonitrile with a K(m) of 40.61 mM. Mass balance studies with [C]acetonitrile indicated that nearly 66% of carbon of acetonitrile was released as CO(2) and 14% was associated with the biomass. Metabolites of acetonitrile in the culture medium were acetic acid and ammonia. The acetate formed in the early stages of growth completely disappeared in the later stages. Cell extracts of acetonitrile-grown cells contained activities corresponding to nitrile hydratase and amidase, which mediate the breakdown of actonitrile into acetic acid and ammonia. Both enzymes were intracellular and inducible and hydrolyzed a wide range of substrates. The specific activity of amidase was at least 150-fold higher than the activity of the enzyme nitrile hydratase. PMID- 16348009 TI - Colloidal Gold Cytochemistry of Endo-1,4-beta-Glucanase, 1,4-beta-D-Glucan Cellobiohydrolase, and Endo-1,4-beta-Xylanase: Ultrastructure of Sound and Decayed Birch Wood. AB - Colloidal gold coupled to endo-1,4-beta-glucanase II (EG II) and 1,4-beta-D glucan cellobiohydrolase I (CBH I), isolated from Trichoderma reesei (QM9414), and endo-1,4-beta-xylanase from Aureobasium pullulans (NRRLY-2311-1) was used successfully to determine the ultrastructural localization of cellulose and xylan in sound birch wood. In addition, these enzyme-gold complexes demonstrated the distribution of cellulose and xylan after decay by three white rot fungi, Phanerochaete chrysosporium, Phellinus pini, and Trametes versicolor, and one brown rot fungus, Fomitopis pinicola. Transverse sections of sound wood showed that EG II was localized primarily on the S(1) layer of the secondary wall, whereas CBH I labeled all layers of the secondary wall. Oblique sections showed a high concentration of gold labeling, using EG II or CBH I. Preference for the sides of the microfibrillar structure was observed for both EG II and CBH I, whereas only CBH I had a specificity for the cut ends of microfibrils. Labeling with the xylanase-gold complex occurred primarily in the inner regions of the S(2) layer, S(1), and the middle lamella. In contrast, little labeling occurred in the middle lamella with EG II or CBH I. Intercellular regions within the cell corners of the middle lamella were less electron dense and labeled positively when EG II- and xylanase-gold were used. Wood decayed by P. chrysosporium or P. pini was delignified, and extensive degradation of the middle lamella was evident. The remaining secondary walls labeled with EG II and CBH I, but little labeling was found with the xylanase-gold complex. Wood decayed by T. versicolor was nonselective, and erosion of all cell wall layers was apparent. Remaining wall layers near sites of erosion labeled with both EG II and CBH I. Erosion troughs that reached the S(1) layer or the middle lamella had less xylanase-gold labeling in the adjacent cell wall that remained. Brown-rotted wood had very low levels of gold particles present in sections treated with EG II or xylanase. Labeling with CBH I had the lowest concentrations in the S(2) layer near cell lumina and corresponded to sites with the most extensive degradation. PMID- 16348010 TI - Purification and Characterization of an Aminopeptidase from Lactococcus lactis subsp. cremoris AM2. AB - An aminopeptidase was purified from cell extracts of Lactococcus lactis subsp. cremoris AM2 by ion-exchange chromatography. After electrophoresis of the purified enzyme in the presence or absence of sodium dodecyl sulfate, one protein band was detected. The enzyme was a 300-kilodalton hexamer composed of identical subunits not linked by disulfide bridges. Activity was optimal at 40 degrees C and pH 7 and was inhibited by classical thiol group inhibitors. The aminopeptidase hydrolyzed naphthylamide-substituted amino acids, as well as dipeptides and tripeptides. Longer protein chains such as the B chain of insulin were hydrolyzed, but at a much slower rate. The Michaelis constant (K(m)) and the maximal rate of hydrolysis (V(max)) were, respectively, 4.5 mM and 3,600 pkat/mg for the substrate l-histidyl-beta-naphthylamide. Amino acid analysis showed that the enzyme contained low levels of hydrophobic residues. The partial N-terminal sequence of the first 19 residues of the mature enzyme was determined. Polyclonal antibodies were obtained from the purified enzyme, and after immunoblotting, there was no cross-reaction between these antibodies and other proteins in the crude extract. PMID- 16348011 TI - Intracellular Location and O(2) Sensitivity of Uptake Hydrogenase in Azospirillum spp. AB - Uptake hydrogenase activity of Azospirillum brasilense in vitro (cell-free extract) was very much more sensitive to O(2) than was that of A. amazonense, and the O(2) pressure optima for uptake hydrogenase activities were 0.01 and 0.4 to 3 kPa for A. brasilense and A. amazonense, respectively. The addition of superoxide dismutase did not increase uptake hydrogenase activity of A. brasilense either in vivo or in vitro. The O(2) uptake rates of A. brasilense and A. amazonense were nearly the same. Inhibition of A. brasilense O(2)-dependent uptake hydrogenase activity by O(2) was highly reversible under the conditions tested. O(2) also markedly inhibited in vitro methylene blue-dependent uptake hydrogenase activity of A. brasilense, and this inhibition was highly reversible. It is concluded that the difference in O(2) tolerance of the uptake hydrogenases is not due to a difference in respiratory protection in the two species and may be due to inherent differences in the two enzymes. For the three species, A. brasilense, A. amazonense, and A. lipoferum, almost all the recovered methylene blue-dependent uptake hydrogenase activity was associated with the membrane fraction. PMID- 16348012 TI - Fungal Growth Stimulation by CO(2) and Root Exudates in Vesicular-Arbuscular Mycorrhizal Symbiosis. AB - Transformed roots of carrot were used to determine the effects of root metabolites on hyphal development from spores of the vesicular-arbuscular mycorrhizal fungus Gigaspora margarita. Hyphal growth of this obligately biotrophic symbiont was greatly stimulated by a synergistic interaction between volatile and exudated factors produced by roots. Root volatiles alone provided little stimulation, and root exudates alone had no effect. For the first time, carbon dioxide was demonstrated to be a critical root volatile involved in the enhancement of hyphal growth. C-labeled root volatiles were fixed by the fungus and thus strongly suggested that CO(2) served as an essential carbon source. PMID- 16348013 TI - Colonization of a Submersed Aquatic Plant, Eurasian Water Milfoil (Myriophyllum spicatum), by Fungi under Controlled Conditions. AB - A laboratory assay to assess colonization of a submersed aquatic plant, Eurasian water milfoil (Myriophyllum spicatum), by fungi was developed and used to evaluate the colonization potential of Colletotrichum gloeosporioides, Acremonium curvulum, Cladosporium herbarum, Aureobasidium pullulans, a Paecilomyces sp., and an unidentified sterile, septate fungus. Stem segments of plants were first immersed in suspensions of fungal propagules for 24 h and then washed to remove all but the tightly attached component of the population. Inoculation was followed by two growth cycles of 3 days each. At the start of each cycle, washed plants were transferred to a mineral salts medium to provide an opportunity for the attached fungal populations to grow. After each growth period, plants were again washed, and fungal populations in the medium (nonattached), loosely attached and tightly attached to the plant, and within the plant (endophytic) were assayed by dilution plating. The fungi differed in the extent to which they attached to water milfoil and in their ability to grow in association with it. There were relatively few significant differences among the tightly attached fungal populations after 24 h, but growth of the better colonizers led to a greater number of significant differences after 4 and 7 days. In addition, the better colonizers showed sustained regrowth of loosely and nonattached fungal propagules in the face of intermittent removal by washing. A milfoil pathogen, C. gloeosporioides, was the only endophytic colonizer; it was also among the best epiphytic colonizers but was not demonstrably better than A. curvulum, a fungus commonly found as an epiphyte on watermilfoil. The yeastlike hyphomycete Aureobasidium pullulans was the only fungus that consistently failed to establish an increasing population on the plant. PMID- 16348014 TI - Selenate reduction to elemental selenium by anaerobic bacteria in sediments and culture: biogeochemical significance of a novel, sulfate-independent respiration. AB - Interstitial water profiles of SeO(4), SeO(3), SO(4), and Cl in anoxic sediments indicated removal of the seleno-oxyanions by a near-surface process unrelated to sulfate reduction. In sediment slurry experiments, a complete reductive removal of SeO(4) occurred under anaerobic conditions, was more rapid with H(2) or acetate, and was inhibited by O(2), NO(3), MnO(2), or autoclaving but not by SO(4) or FeOOH. Oxidation of acetate in sediments could be coupled to selenate but not to molybdate. Reduction of selenate to elemental selenium was determined to be the mechanism for loss from solution. Selenate reduction was inhibited by tungstate and chromate but not by molybdate. A small quantity of the elemental selenium precipitated into sediments from solution could be resolublized by oxidation with either nitrate or FeOOH, but not with MnO(2). A bacterium isolated from estuarine sediments demonstrated selenate-dependent growth on acetate, forming elemental selenium and carbon dioxide as respiratory end products. These results indicate that dissimilatory selenate reduction to elemental selenium is the major sink for selenium oxyanions in anoxic sediments. In addition, they suggest application as a treatment process for removing selenium oxyanions from wastewaters and also offer an explanation for the presence of selenite in oxic waters. PMID- 16348015 TI - Simulation of airborne microbial droplet transport. AB - The framework for a simulation model which describes the dispersion of individual droplets of water containing viable microbes is presented. The model accounts for physical, chemical, biological, and measured meteorological parameters of each droplet at each of many short time steps. Repeating the modeling process for many droplets will simulate a cloud of droplets. The model is compared with the Tulelake, Calif., release in 1988 and found to show very similar patterns of deposition within 30 m (the maximum observation distance of the source. A hypothesis for the survival sequence in the microbe-containing droplets is discussed. PMID- 16348016 TI - Detection, growth, and amine-producing capacity of lactobacilli in cheese. AB - A differential plating medium was developed to detect decarboxylating lactobacilli in cheese. With this medium, 15 cheeses made from raw milk were investigated for the presence of these bacteria. Five histidine-decarboxylating strains and one tyrosine-decarboxylating strain were isolated. The isolates were identified with the API 50L system. Accordingly, each of the five histidine decarboxylating strains was identified as Lactobacillus buchneri, whereas the tyrosine-decarboxylating strain is a representative of Lactobacillus brevis. Cheesemaking experiments using a low inoculum concentration of the histidinedecarboxylating L. buchneri strain St2A (0.2 CFU/ml of milk) showed that, under conditions of accelerated proteolysis, histamine may accumulate rapidly; after 3 months of ripening, 410 mg/kg was found. An inoculum concentration of 5 CFU/ml gave rise to the formation of 1,060 mg/kg. PMID- 16348017 TI - Physiological and Morphological Modifications in Immobilized Gibberella fujikuroi Mycelia. AB - Constraints created by immobilization conditions modified the physiological behavior and morphological characteristics of Gibberella fujikuroi mycelia in comparison with their development in free-cell conditions. G. fujikuroi mycelia were immobilized in different support matrices (polyurethane, carrageenan, and alginate) and showed a variety of reactions in response to the different microenvironmental factors encountered during and after immobilization. The best support with respect to gibberellic acid yield and biocatalyst stability was found to be an alginate with a high degree of polymerization. The most visible effects of immobilization included changes in growth development, morphological appearance, metabolite production, mycelial pigmentation, mycelial viability under starvation conditions, and induction of resting forms when previously immobilized mycelia were subcultured. PMID- 16348019 TI - Epiphytic Occurrence of Azorhizobium caulinodans and Other Rhizobia on Host and Nonhost Legumes. AB - A large population of Azorhizobium caulinodans was present on Sesbania rostrata; up to 5 x 10 cm were found on leaves and fewer were found on flowers. Although A. caulinodans was also present on the leaves of Sesbania aculeata (nonhost), the populations were much smaller than that observed on S. rostrata. The population of S. aculeata rhizobia on host leaves was less than 30 cm, and their presence on host flowers was sporadic. Aeschynomene afraspera and Aeschynomene aspera rhizobia, which are profusely stem nodulating, were found on the leaves of host and nonhost plants and on the flowers of host plants, but, Aeschynomene pratensis and Aeschynomene sensitiva rhizobia were not found on the leaves and flowers of host plants. PMID- 16348018 TI - Metabolism of Ferulic Acid by Paecilomyces variotii and Pestalotia palmarum. AB - Ferulic acid metabolism was studied in cultures of two micromycetes producing different amounts of phenol oxidases. In cultures of the low phenol oxidase producer Paecilomyces variotii, ferulic acid was decarboxylated to 4 vinylguaiacol, which was converted to vanillin and then either oxidized to vanillic acid or reduced to vanillyl alcohol. Vanillic acid underwent simultaneously an oxidative decarboxylation to methoxyhydroquinone and a nonoxidative decarboxylation to guaiacol. Methoxyhydroquinone and guaiacol were demethylated to yield hydroxyquinol and catechol, respectively. Catechol was hydroxylated to pyrogallol. Degradation of ferulic acid by Paecilomyces variotii proceeded mainly via methoxyhydroquinone. The high phenol oxidase producer Pestalotia palmarum catabolized ferulic acid via 4-vinylguaiacol, vanillin, vanillyl alcohol, vanillic acid, and methoxyhydroquinone. However, the main reactions observed with this fungus involved polymerization reactions. PMID- 16348020 TI - Microbiological comparison of surface soil and unsaturated subsurface soil from a semiarid high desert. AB - Thirty-two chemoheterotrophic bacteria were isolated from unsaturated subsurface soil samples obtained from ca. 70 m below land surface in a high desert in southeastern Idaho. Most isolates were gram positive (84%) and strict aerobes (79%). Acridine orange direct counts of microbes in one subsurface sample showed lower numbers than similar counts performed on surface soils from the same location (ca. 5 x 10 versus 2 x 10 cells per g [dry weight] of soil), but higher numbers than those from plate counts performed on the subsurface material. Another sample taken from the same depth at another location showed no evidence of colonies under identical conditions. Soil analyses indicated that subsurface sediments versus surface soils were slightly alkaline (pH 7.9 versus 7.4), had a higher water content (25.7 versus 6.3%), and had lower organic carbon concentrations (0.05 to 0.17 versus 0.25% of soil dry weight). Analyses of biologically relevant gases from the unsaturated subsurface indicated an aerobic environment. As in other unsaturated soil environments, either a high proportion of bacteria in these subsurface sediments are not viable or they are incapable of growth on conventional media under aerobic conditions. The presence and numbers of bacteria in these deep sediments may be influenced by colonization opportunities afforded by periodic percolation of surface water through fractures in overlying strata. PMID- 16348022 TI - Worldwide Abundance and Distribution of Bacillus thuringiensis Isolates. AB - We found the insect control agent Bacillus thuringiensis to be a ubiquitous soil microorganism. Using acetate selection to screen soil samples, we isolated B. thuringiensis in 785 of 1,115 soil samples. These samples were obtained in the United States and 29 other countries. A total of 48% of the B. thuringiensis isolates (8,916 isolates) fit the biochemical description of known varieties, while 52% represented undescribed B. thuringiensis types. Over 60% (1,052 isolates) of the isolates tested for toxicity were toxic to insects in the orders Lepidoptera or Diptera. Soil samples were collected from various habitats, including those habitats with different numbers of insects. The current presence of insects did not predict the presence of B. thuringiensis in a particular soil sample. B. thuringiensis was most abundant in samples from Asia. PMID- 16348021 TI - Growth and Nodulation Responses of Rhizobium meliloti to Water Stress Induced by Permeating and Nonpermeating Solutes. AB - Isolates of Rhizobium meliloti, representing antigenically distinct indigenous serogroups 31 and 17, were grown in yeast extract-mannitol broth (YEM) containing NaCl or polyethylene glycol (PEG) to provide external water potentials ranging from -0.15 to -1.5 MPa. Several differences were found between representatives of the two groups in their abilities to adapt to water stress induced by the nonpermeating solute PEG. At potentials below -0.5 MPa, strain 31 had a lower specific growth rate than strain 17 and an irregular cell morphology. In contrast, neither growth nor cell morphology of either strain was affected significantly over the same range of water potentials created by a permeating solute, NaCl. Despite the superior growth of strain 17 at the low water potentials imposed by PEG, upshock of water-stressed cells (-1.0 MPa; PEG) into normal YEM (-0.15 MPa) resulted in a faster recovery of growth by strain 31 than by strain 17. Different responses of the two strains to a water potential increase were also revealed in nodulation studies. Strain 31 required significantly fewer days to nodulate alfalfa than strain 17 did when the strains were transferred from YEM with PEG at -1.0 MPa onto the roots of alfalfa seedlings in plant growth medium (-0.1 MPa). The addition of supplemental calcium (0.1 mM) to growth medium with PEG (-1.0 MPa) reduced the differences between strains in their responses to water stress. The severe growth restriction and morphological abnormalities shown by strain 31 were corrected, and the prolonged recovery time shown by water-stressed cells (-1.0 MPa; PEG) of strain 17 upon transfer to normal YEM was shortened. The latter strain also nodulated earlier and more rapidly after growth in PEG medium at -1.0 MPa in the presence of supplemental calcium ions. These results indicate that the efficacy of osmoregulation can vary among strains of the same species and that the mechanism of osmoregulation may differ depending on the nature of the water stress. PMID- 16348023 TI - Alternative Environmental Roles for Cellulose Produced by Acetobacter xylinum. AB - The cellulose-producing bacterium Acetobacter xylinum has been considered a strict aerobe, and it has been suggested that the function of cellulose is to hold cells in an aerobic environment. In this study, we showed that A. xylinum is capable of growing microaerophilically. Cellulose pellicles provided significant protection to A. xylinum cells from the killing effects of UV light. In experiments measuring colonization by A. xylinum, molds, and other bacteria on pieces of apple, cellulose pellicles enhanced colonization of A. xylinum on the substrate and provided protection from competitors which use the same substrate as a source of nutrients. Cellulose pellicles produced by A. xylinum may have multiple functions in the growth and survival of the organism in nature. PMID- 16348024 TI - Effect of growth conditions and trehalose content on cryotolerance of bakers' yeast in frozen doughs. AB - The cryotolerance in frozen doughs and in water suspensions of bakers' yeast (Saccharomyces cerevisiae) previously grown under various industrial conditions was evaluated on a laboratory scale. Fed-batch cultures were very superior to batch cultures, and strong aeration enhanced cryoresistance in both cases for freezing rates of 1 to 56 degrees C min. Loss of cell viability in frozen dough or water was related to the duration of the dissolved-oxygen deficit during fed batch growth. Strongly aerobic fed-batch cultures grown at a reduced average specific rate (mu = 0.088 h compared with 0.117 h) also showed greater trehalose synthesis and improved frozen-dough stability. Insufficient aeration (dissolved oxygen deficit) and lower growth temperature (20 degrees C instead of 30 degrees C) decreased both fed-batch-grown yeast cryoresistance and trehalose content. Although trehalose had a cryoprotective effect in S. cerevisiae, its effect was neutralized by even a momentary lack of excess dissolved oxygen in the fed-batch growth medium. PMID- 16348025 TI - Production of Leuconostoc oenos Biomass under pH Control. AB - Leuconostoc oenos was grown on apple juice-based media. The effect of pH control on metabolism and biomass production was studied. Without pH control, L. oenos acidified the apple juice media to approximately pH 3.6. More than 75% of the malic acid was used under these conditions, but less than half of the carbohydrates was assimilated. Under pH control, biomass yields increased by 60%; most of the malic acid was used, but high levels of unfermented carbohydrates remained. The addition of tomato juice, vitamins, nucleotides, Mn, and malic acid did not permit further increases in the cell counts; however, malic acid did induce further acidification. Growth without pH control favored a more homofermentative metabolism. Biomass production was higher in filter-sterilized apple juice media compared with that in the autoclaved media. PMID- 16348026 TI - Bradyrhizobium japonicum Inoculant Mobility, Nodule Occupancy, and Acetylene Reduction in the Soybean Root System. AB - In the American Midwest, superior inoculant rhizobia applied to soybeans usually occupy only 5 to 20% of nodules, and response to inoculation is the exception rather than the rule. Attempts to overcome this problem have met with limited success. We evaluated the ability of Bradyrhizobium japonicum, supplied as a seed coat inoculant, to stay abreast of the infectible region of the developing soybean root system. The rhizoplane population of the inoculant strain declined with distance from site of placement, the decrease being more pronounced on lateral than on taproots. This decline was paralleled by a decrease in inoculant strain nodule occupancy. Inoculant bradyrhizobia contributed little to nodulation of lateral roots, which at pod-fill accounted for more than 50% of nodule number and mass, and were major contributors to acetylene reduction activity. From these data, it appears that inoculant bradyrhizobia are competitive with indigenous soil strains at the point of placement in the soil but have limited mobility and so are incapable of sustaining high populations throughout the developing root system. The result is low nodule occupancy by the inoculant strain in the tapand lateral roots. Future studies should address aspects of inoculant placement and establishment. PMID- 16348027 TI - Reinvestigation of Brevibacterium sp. Strain KY-4313 as a Source of Canthaxanthin. AB - The hydrocarbon-utilizing Brevibacterium sp. strain KY-4313 was reevaluated for its potential to produce canthaxanthin, a carotenoid pigment of strong commercial interest. Three approaches were used to optimize the canthaxanthin yield from this organism, i.e., the preparation of mutants, the addition of supposedly carotenogenic chemicals to the growth medium, and growth promotion. Following treatment of the parent strain with N-nitrosomethylurea, a presumed mutant was isolated which showed a 32% increase in cellular canthaxanthin content. No effective carotenogenic chemicals were found in connection with hydrocarbon fermentations, in which mainly growth promotion through periodic medium renewal proved conducive to enhanced pigment production. Carotenogenesis could be stimulated in brain heart infusion broth by adding alcohols or retinol. Improved growth in this medium was generally not associated with higher canthaxanthin yields. Both superior growth and pigment levels were obtained in a newly designed medium based on fumaric acid-molasses. The maximum yields of canthaxanthin in shake flasks were (in milligrams per liter) 4.2 (brain heart infusion broth plus propanol-zinc sulfate), 3.6 (hydrocarbon medium), and 9.3 (fumaric acid molasses), which represent a significant improvement over the originally reported optimal result (1 mg/liter). The corresponding yields of echinenone, the direct precursor of canthaxanthin, were 1.2, 1.6, and 2.3 mg/liter, respectively. Two liter hydrocarbon batch fermentations involving medium renewal maximally produced 7.2 mg of canthaxanthin and 3.7 mg of echinenone per liter. PMID- 16348028 TI - Alkaline Phosphatase and Other Hydrolyases Produced by Cenococcum graniforme, an Ectomycorrhizal Fungus. AB - Cell extracts of Cenococcum graniforme have been found to contain the following hydrolytic enzymes: protease, esterase, alpha-d-galactopyranosidase, beta-d galactopyranosidase, alpha-d-mannopyranosidase, beta-d-xylopyranosidase, alpha-d glucopyranosidase, beta-d-glucopyranosidase, and alkaline phosphatase. Sulfatase, inorganic pyrophosphatase, and beta-d-mannopyranosidase were not detected in the extracts. beta-d-Xylopyranosidase and alpha-d-mannopyranosidase were most active in the neutral pH range, protease and phosphatase were most active in the alkaline pH range, and other enzymes were most active in the acidic pH range. These enzymes showed a high association with cell wall material, and the release of enzymes from the cells into the culture fluid appeared to occur only when the cells were undergoing autolysis. Alkaline phosphatase in C. graniforme is a constitutive enzyme, and examination of the alkaline phosphatase following a purification of 265-fold produced the following characteristics: pH optimum of 9.5, M(r) of 60,000, K(m) of 2.1 x 10 M for p-nitrophenylphosphate, and activation energy for hydrolysis of the substrate at 9.9 kcal (1 cal = 4.184 J)/mol. PMID- 16348029 TI - Host Plant Effects on Nodulation and Competitiveness of the Bradyrhizobium japonicum Serotype Strains Constituting Serocluster 123. AB - Strains in Bradyrhizobium japonicum serocluster 123 are the major indigenous competitors for nodulation in a large portion of the soybean production area of the United States. Serocluster 123 is defined by the serotype strains USDA 123, USDA 127, and USDA 129. The objective of the work reported here was to evaluate the ability of two soybean genotypes, PI 377578 and PI 417566, to restrict the nodulation and reduce the competitiveness of serotype strains USDA 123, USDA 127, and USDA 129 in favor of the highly effective strain CB1809 and to determine how these soybean genotypes alter the competitive relationships among the three serotype strains in the serocluster. The soybean genotypes PI 377578 and PI 417566 along with the commonly grown cultivar Williams were planted in soil essentially free of soybean rhizobia and inoculated with single-strain treatments of USDA 123, USDA 127, USDA 129, or CB1809 and six dual-strain competition treatments of USDA 123, USDA 127, or USDA 129 versus CB1809, USDA 123 versus USDA 127, USDA 123 versus USDA 129, and USDA 127 versus USDA 129. PI 377578 severely reduced the nodulation and competitiveness of USDA 123 and USDA 127, while PI 417566 similarly affected the nodulation and competitiveness of USDA 129. Thus, the two soybean genotypes can reduce the nodulation and competitiveness of each of the three serocluster 123 serotype strains. Our results indicate that host control of restricted nodulation and reduced competitiveness is quite specific and effectively discriminates between B. japonicum strains which are serologically related. PMID- 16348030 TI - Development of a solid medium for growth and isolation of axenic microcystis strains (cyanobacteria). AB - Solid media on a base of B-12 or CB medium with agarose or agarose of low melting temperature were developed for the cultivation of Microcystis species. The media with 0.4% gel showed the highest number of CFU, and increasing the gel concentration resulted in a reduction of the number of CFU. There was no difference in the numbers of CFU between pour and spread plates made of the solid media. By using the solid media, 31 clones of Microcystis species were isolated from natural blooms in Lake Kasumigaura, and 5 axenic strains (1 of M. wesenbergii and 4 of M. aeruginosa) were established from the clones. PMID- 16348031 TI - Synergistic Competitive Inhibition of Ferrous Iron Oxidation by Thiobacillus ferrooxidans by Increasing Concentrations of Ferric Iron and Cells. AB - Oxidation of ferrous iron by Thiobacillus ferrooxidans SM-4 was inhibited competitively by increasing concentrations of ferric iron or cells. A kinetic analysis showed that binding of one inhibitor did not exclude binding of the other and led to synergistic inhibition by the two inhibitors. Binding of one inhibitor, however, was affected by the other inhibitor, and the apparent inhibition constant increased with increasing concentrations of the other inhibitor. PMID- 16348032 TI - Corky root of lettuce caused by strains of a gram-negative bacterium from muck soils of Florida, new york, and wisconsin. AB - Slow-growing bacteria similar to the bacterium causing lettuce corky root (CR) in California (strain CA1) were isolated from muck soils of Florida, New York, and Wisconsin, using lettuce seedlings as bait. All strains were tested for reaction with polyclonal antibodies produced against strain CA1 and for pathogenicity on CR-susceptible (Salinas) and CR-resistant (Green Lake) lettuce cultivars in a greenhouse. Five strains from Florida, three from New York, and three from Wisconsin induced severe CR symptoms on Salinas and mild symptoms on Green Lake. All strains were gram-negative, aerobic, oxidase positive, and catalase positive and reduced nitrate to ammonia. Whole-cell fatty acid compositions were similar for all strains and resembled that of Pseudomonas paucimobilis. Since this fatty acid pattern is unique, it is suggested that CR of lettuce is caused by strains of the same bacterium in Florida, New York, Wisconsin, and California. PMID- 16348033 TI - Identification of Mycelium-Associated Cellulase from Streptomyces reticuli. AB - Among 180 Streptomyces strains tested, 25 were capable of hydrolyzing microcrystalline cellulose (Avicel) at 30 degrees C. Streptomyces reticuli was selected for further studies because of its ability to grow at between 30 and 50 degrees C on Avicel. Enzymatic activities degrading Avicel, carboxymethyl cellulose, and cellobiose were found both in the culture supernatant and in association with the mycelium and crystalline substrate. The bound enzymes were efficiently solubilized by repeated washes with buffer of low ionic strength (50 mM Tris hydrochloride [pH 7.5]) and further purified by fast protein liquid chromatography. A high-molecular-weight Avicelase of >300 kilodaltons could be separated from carboxymethyl cellulase (CMCase) and beta-glucosidase activities (molecular mass, 40 to 50 kilodaltons) by gel filtration on Superose 12. The CMCase fraction was resolved by Mono Q anion-exchange chromatography into two enzymes designated CMCase 1 and CMCase 2. The beta-glucosidase activity was found to copurify with CMCase 2. The purified cellulase components showed optimal activity at around pH 7.0 and temperatures of between 45 and 50 degrees C. Avicelase (but not CMCase) activity was stimulated significantly by the addition of CaCl(2). PMID- 16348034 TI - Differential Expression of Xylanases and Endoglucanases in the Hybrid Derived from Intergeneric Protoplast Fusion between a Cellulomonas sp. and Bacillus subtilis. AB - A stable hybrid obtained by protoplast fusion between a Cellulomonas sp. and Bacillus subtilis exhibits an altered pattern of enzyme induction with different cellulosic substrates. Unlike in the Cellulomonas sp., xylanase was induced in the hybrid organism specifically by xylan, and endoglucanase was induced by carboxymethyl cellulose. The amount and specific activity of xylanase produced by the hybrid were more than those produced by the Cellulomonas sp. beta-Glucosidase which is cell bound or intracellular in the Cellulomonas sp. was secreted by the hybrid organism, and relative amounts of extracellular beta-glucosidase were high. Furthermore, this extracellular beta-glucosidase activity was dependent on the nature of the cellulosic substrate. Endoglucanases synthesized in the hybrid differed in their electrophoretic mobilities as compared with the parental enzymes. PMID- 16348035 TI - Peptide Utilization Encoded by Lactococcus lactis subsp. lactis SSL135 Chromosomal DNA. AB - A cloned chromosomal fragment of Lactococcus lactis subsp. lactis SSL135 on plasmid p VS8 in an L. lactis subsp. lactis MG1614 background enabled proteinase negative strain MG1614 to grow in autoclaved milk. The strain (VS230) did not, however, degrade milk proteins and did not grow in pasteurized milk. In contrast, a strain (VS150) carrying p VS9, the proteinase plasmid of SSL135, in an MG1614 background degraded beta-casein but did not grow in milk. VS230 was shown to utilize peptides produced by VS150 in growth experiments in pasteurized milk preincubated with the latter strain. The peptide utilization phenotype linked with p VS8 was further confirmed by growth of VS230 on tryptic peptide fractions, on which the plasmid-free but otherwise isogenic strain MG1614 failed to grow. Plasmid p VS8 produced 69-, 42-, 38-, and 36-kilodalton proteins, as determined by in vitro transcription-translation. At least three of these proteins affected the peptide utilization phenotype. We suggest that there could be a coupled peptidase-peptide transport system encoded by the chromosomal fragment. PMID- 16348036 TI - Localization of Separate Genetic Loci for Reduced Sensitivity towards Small Isometric-Headed Bacteriophage sk1 and Prolate-Headed Bacteriophage c2 on pGBK17 from Lactococcus lactis subsp. lactis KR2. AB - The mechanism of reduced sensitivity to the small isometric-headed bacteriophage sk1 encoded on a 19-kilobase (kb) HpaII fragment subcloned from pKR223 of Lactococcus lactis subsp. lactis KR2 was examined. The reduced sensitivity to phage sk1 was due to a modest restriction/modification (R/M) system that was not active against prolate-headed phage c2. The genetic loci for the R/M system against sk1 and the abortive phage infection (Abi) mechanism effective against phage c2 were then localized by restriction mapping, subcloning, and deletion analysis. The restriction gene was localized to a region of a 2.7-kb EcoRV fragment and included an EcoRI site within that fragment. The modification gene was found to be physically separable from the restriction gene and was present on a 1.75-kb BstEII-XbaI fragment. The genetic locus for the Abi phenotype against phage c2 was localized to a region containing a 1.3-kb EcoRI fragment. Attempts to clone the c2 Abi mechanism independent of the sk1 R/M system were unsuccessful, suggesting that expression of the abi genes required sequences upstream of the modification gene. Some pGBK17 (vector pGB301 plus a 19-kb HpaII insert fragment) transformants exhibited the R/M system against phage sk1 but lost the Abi mechanism against phage c2. These transformants contained a 1.2- to 1.3-kb insertion in the Abi region. The data identified genetic loci on a cloned 19-kb HpaII fragment responsible for restriction activity and for modification activity against a small isometric-headed phage and for Abi activity against prolate-headed phage c2. A putative insertion element was also found to inactivate the abi gene(s). PMID- 16348037 TI - Effect of growth rate and starvation-survival on cellular DNA, RNA, and protein of a psychrophilic marine bacterium. AB - DNA, RNA, and protein concentrations from starved ANT-300 cell populations grown at different growth rates fluctuated corresponding to the three stages of starvation-survival on total and viable cell bases. During stage 1 of starvation survival, two to three peaks in the concentration levels for all three macromolecules were characteristic. During stage 2, DNA per total cell dropped to between 4.2 and 8.3% of the original amount for all of the cell populations examined, and it stabilized throughout stage 3. The decrease in DNA per cell was also observed in electron micrographs of cellular DNA in unstarved compared with starved cells. The fluctuations of RNA and protein per total cell concentrations observed during stage 2 coincided in all cases, except for the cells from dilution rate (D) = 0.015 h. This ANT-300 cell population showed a decrease in RNA per total cell to only 29.2% and an increase in protein to 129.7% of the original amount after 98 days of starvation. During stage 3, DNA, RNA, and protein concentrations per total cell also stabilized to continuous levels. Cells from the faster-growth-rate cell populations of D = 0.170 h and batch culture had elevated protein per total cell concentrations, which remained primarily residual during the starvation period. Starved cells from D = 0.015 h had estimated nucleoid and cell volumes of 0.018 and 0.05 mum, respectively, yielding a nucleoid volume/cell volume ratio of 0.40. We consider these data to indicate that slow-growth-rate cells are better adapted for starvation-survival than their faster-growth-rate counterparts. PMID- 16348038 TI - Iron-binding compounds produced by wood-decaying basidiomycetes. AB - The chrome azurol-S universal siderophore assay and the rapid paper electrophoresis siderophore assay were used to screen 10 wood-decaying basidiomycete isolates for the formation of iron-chelating compounds. All 10 isolates were positive for chrome azurol-S reactivity on solid plating medium and in liquid cultures, and 9 of the 10 isolates produced fluorescent iron-binding compounds in the paper electrophoresis assay. PMID- 16348039 TI - Toxicity of Trichloroethylene to Pseudomonas putida F1 Is Mediated by Toluene Dioxygenase. AB - Trichloroethylene was metabolically activated by toluene dioxygenase to produce toxic effects in Pseudomonas putida F1. Cytotoxicity was indicated by growth inhibition and by the covalent modification of cellular molecules in P. putida F1 exposed to [C]trichloroethylene. With a toluene dioxygenase mutant, neither growth inhibition nor alkylation of intracellular molecules was observed. PMID- 16348040 TI - Electron Spin Resonance Analysis of the Effect of Butanol on the Membrane Fluidity of Intact Cells of Clostridium acetobutylicum. AB - Analysis of electron spin resonance spectra of 5-doxyl stearic acid in aqueous suspensions of Clostridium acetobutylicum ATCC 824 and the butanol-tolerant SA-2 derivative during a small-scale fermentation at three different butanol challenge levels indicated that the SA-2 strain is able to respond to the physical fluidizing effect of high (1.5%) butanol challenge by reducing its membrane fluidity at 12 and 30 h. The wild-type 824 strain was unable to so respond when challenged at the 1.5% level. PMID- 16348041 TI - Simple method for determination of biodegradable dissolved organic carbon in water. AB - Determination of biodegradable dissolved organic carbon in waters is of particular importance for the water treatment industry. A simple method for determining biodegradable dissolved organic carbon which is applicable to surface and drinking water is proposed. It consists of sterilizing the water sample, inoculating it with autochthonous bacteria, and measuring the decrease in dissolved organic carbon concentration due to the carbon oxidization by bacteria. The detailed experimental procedure is discussed, and validation of the method is presented. The method has been used for studying river waters and for drinking water treatment plant design. PMID- 16348042 TI - Inducible Secretion of a Cellulase from Clostridium thermocellum in Bacillus subtilis. AB - A host-vector system for inducible secretion during the logarithmic growth phase in Bacillus subtilis has been developed. The B. subtilis levansucrase gene promoter and the region encoding its signal sequence have been used. The endoglucanase A of Clostridium thermocellum was used as a model protein to test the efficiency of the system. Effective inducible secretion of the endoglucanase A was observed when either the levansucrase signal sequence or its own signal sequence was used. Expression of the endoglucanase A in different genetic backgrounds of B. subtilis showed that its regulation was similar to that of levansucrase, and high enzyme activity was recovered from the culture supernatant of a hyperproducing B. subtilis sacU(Hy) strain. The molecular weight of 46,000 estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the secreted endoglucanase A is compatible with the calculated molecular weight of the mature polypeptide. PMID- 16348043 TI - Rhizobacteria of cotton and their repression of seedling disease pathogens. AB - During the 1983 field season, the rhizobacteria (including organisms from rhizosphere soil and the root rhizoplane) of cotton plants at one location in Mississippi were inventoried at different plant growth stages. Isolates (1,000) were identified to the genus level and characterized for repression of Pythium ultimum and Rhizoctonia solani. Cotton seedlings were initially colonized by bacteria of many different genera, and populations quickly reached 10 CFU/g of root tissue. As the season progressed, the bacterial populations declined as root mass increased and the roots became more woodlike in consistency. Fluorescent pseudomonads were the most numerous gram-negative rhizobacterial isolates of those that were randomly collected and identified, and they provided the largest number of isolates with fungal repressive activity. Several other gram-negative bacterial genera were recovered throughout the growing season, and some gram positive bacteria were also isolated routinely, but at lower numbers. There was no correlation between the proportion of rhizobacterial isolates that possessed fungal repressive activity and the plant growth stage from which the isolates were obtained. Approximately twice as many bacterial isolates demonstrated fungal repression in the agar assay compared with the inplanta assay, and isolates were found more frequently with fungal repressive activity against P. ultimum than against R. solai. PMID- 16348044 TI - Specificity of Cellular DNA-Binding Sites of Microbial Populations in a Florida Reservoir. AB - The substrate specificity of the DNA-binding mechanism(s) of bacteria in a Florida reservoir was investigated in short- and long-term uptake studies with radiolabeled DNA and unlabeled competitors. Thymine oligonucleotides ranging in size from 2 base pairs to 19 to 24 base pairs inhibited DNA binding in 20-min incubations by 43 to 77%. Deoxynucleoside monophosphates, thymidine, and thymine had little effect on short-term DNA binding, although several of these compounds inhibited the uptake of the radiolabel from DNA in 4-h incubations. Inorganic phosphate and glucose-1-phosphate inhibited neither short- nor long-term binding of [H]- or [P]DNA, indicating that DNA was not utilized as a phosphorous source in this reservoir. RNA inhibited both short- and long-term radiolabeled DNA uptake as effectively as unlabeled DNA. Collectively these results indicate that aquatic bacteria possess a generalized nucleic acid uptake/binding mechanism specific for compounds containing phosphodiester bonds and capable of recognizing oligonucleotides as short as dinucleotides. This binding site is distinct from nucleoside-, nucleotide-, phosphomonoester-, and inorganic phosphate-binding sites. Such a nucleic acid-binding mechanism may have evolved for the utilization of extracellular DNA (and perhaps RNA), which is abundant in many marine and freshwater environments. PMID- 16348045 TI - Microbial utilization of naturally occurring hydrocarbons at the guaymas basin hydrothermal vent site. AB - The Guaymas Basin (Gulf of California; depth, 2,000 m) is a site of hydrothermal activity in which petroliferous material is formed by thermal alteration of deposited planktonic and terrestrial organic matter. We investigated certain components of these naturally occurring hydrocarbons as potential carbon sources for a specific microflora at these deep-sea vent sites. Respiratory conversion of [1-C]hexadecane and [1(4,5,8)-C]naphthalene to CO(2) was observed at 4 degrees C and 25 degrees C, and some was observed at 55 degrees C, but none was observed at 80 degrees C. Bacterial isolates were capable of growing on both substrates as the sole carbon source. All isolates were aerobic and mesophilic with respect to growth on hydrocarbons but also grew at low temperatures (4 to 5 degrees C). These results correlate well with previous geochemical analyses, indicating microbial hydrocarbon degradation, and show that at least some of the thermally produced hydrocarbons at Guaymas Basin are significant carbon sources to vent microbiota. PMID- 16348046 TI - Sequential photochemical and microbial degradation of organic molecules bound to humic Acid. AB - We studied the effects of photochemical processes on the mineralization by soil microorganisms of [2-C]glycine bound to soil humic acid. Microbial mineralization of these complexes in the dark increased inversely with the molecular weight of the complex molecules. Sunlight irradiation of glycine-humic acid complexes resulted in loss of absorbance in the UV range and an increase in the amount of C labeled low-molecular-weight photoproducts and the rate and extent of mineralization. More than half of the radioactivity in the low-molecular-weight photoproducts appears to be associated with carboxylic acids. Microbial mineralization of the organic carbon increased with solar flux and was proportional to the loss of A(330). Mineralization was proportional to the percentage of the original complex that was converted to low-molecular-weight photoproducts. Only light at wavelengths below 380 nm had an effect on the molecular weight distribution of the products formed from the glycine-humic acid complexes and on the subsequent microbial mineralization. Our results indicate that photochemical processes generate low-molecular-weight, readily biodegradable molecules from high-molecular-weight complexes of glycine with humic acid. PMID- 16348047 TI - Metabolism of Styrene Oxide and 2-Phenylethanol in the Styrene-Degrading Xanthobacter Strain 124X. AB - Styrene oxide and 2-phenylethanol metabolism in the styrene-degrading Xanthobacter sp. strain 124X was shown to proceed via phenylacetaldehyde and phenylacetic acid. In cell extracts 2-phenylethanol was oxidized by a phenazine methosulfate-dependent enzyme, probably a pyrroloquinoline quinone enzyme. Xanthobacter sp. strain 124X also contains a novel enzymatic activity designated as styrene oxide isomerase. Styrene oxide isomerase catalyzes the isomerization of styrene oxide to phenylacetaldehyde. The enzyme was partially purified and shown to have a very high substrate specificity. Of the epoxides tested, styrene oxide was the only substrate transformed. The initial step in styrene metabolism in Xanthobacter sp. strain 124X is oxygen dependent and probably involves oxidation of the aromatic nucleus. PMID- 16348048 TI - Nitrate reduction in a groundwater microcosm determined by N gas chromatography mass spectrometry. AB - Aerobic and anaerobic groundwater continuous-flow microcosms were designed to study nitrate reduction by the indigenous bacteria in intact saturated soil cores from a sandy aquifer with a concentration of 3.8 mg of NO(3)-N liter. Traces of NO(3) were added to filter-sterilized groundwater by using a Darcy flux of 4 cm day. Both assimilatory and dissimilatory reduction rates were estimated from analyses of N(2), N(2)O, NH(4), and N-labeled protein amino acids by capillary gas chromatography-mass spectrometry. N(2) and N(2)O were separated on a megabore fused-silica column and quantified by electron impact-selected ion monitoring. NO(3) and NH(4) were analyzed as pentafluorobenzoyl amides by multiple-ion monitoring and protein amino acids as their N-heptafluorobutyryl isobutyl ester derivatives by negative ion-chemical ionization. The numbers of bacteria and their [methyl-H]thymidine incorporation rates were simultaneously measured. Nitrate was completely reduced in the microcosms at a rate of about 250 ng g day. Of this nitrate, 80 to 90% was converted by aerobic denitrification to N(2), whereas only 35% was denitrified in the anaerobic microcosm, where more than 50% of NO(3) was reduced to NH(4). Assimilatory reduction was recorded only in the aerobic microcosm, where N appeared in alanine in the cells. The nitrate reduction rates estimated for the aquifer material were low in comparison with rates in eutrophic lakes and coastal sediments but sufficiently high to remove nitrate from an uncontaminated aquifer of the kind examined in less than 1 month. PMID- 16348049 TI - Genetic and Biochemical Characterization of Mutations Affecting the Ability of the Yeast Pachysolen tannophilus To Metabolize d-Xylose. AB - Induced mutants, selected for their defective growth on d-xylose while retaining the ability to grow normally on d-glucose, were studied in Pachysolen tannophilus, a yeast capable of converting d-xylose to ethanol. Fourteen of the mutations were found to occur at nine distinct loci, and data indicated that many more loci remain to be detected. Most of the mutations were pleiotropic in character, and the expression of some of them was much affected by nutritional conditions and by genetic background. Mutations at several loci resulted in poor growth on at least one compound that was either an intermediate of the tricarboxylic acid cycle, succinate or alpha-ketoglutarate, or on compounds metabolizable via this cycle, ethanol or glycerol. An initial biochemical characterization of the mutants was undertaken. Analysis for xylose reductase, xylitol dehydrogenase, and xylulose kinase activity showed that one or more of these activities was affected in 12 of 13 mutants. However, drastic reduction in activity of a single enzyme was confined to that of xylitol dehydrogenase by mutations at three different loci and to that of d-xylose reductase by mutation at another locus. Growth of these latter four mutants was normal on all carbon sources tested that were not five-carbon sugars. PMID- 16348050 TI - Physiological Properties of a Mutant of Pachysolen tannophilus Deficient in NADPH Dependent d-Xylose Reductase. AB - A d-xylose reductase mutant of Pachysolen tannophilus was isolated on the basis of its poor growth on d-xylose but normal growth on xylitol and d-glucose. Fractionation of cell extracts indicated that the mutant was deficient in d xylose reductase activity that used NADPH exclusively as a cofactor, but not in activity that used both NADH and NADPH. Mutant cultures grown on d-xylose as the sole carbon source exhibited some properties that would be desired in improved strains. Growth rate, growth yield, and d-xylose consumption rate of the mutant were less sensitive than those of the wild type to changes in aeration rate. d Xylose was utilized more efficiently in that less of a by-product, xylitol, was produced. In addition, under low aeration conditions, more ethanol was produced. A disadvantage was a relatively slow rate of d-xylose utilization. PMID- 16348051 TI - Efficacy of phospholipid analysis in determining microbial biomass in sediments. AB - Improvements in the analysis of lipid-bound phosphates resulted in a simplified and sensitive method for determining microbial biomass in sediments. Sensitivity was enhanced over previous methods by use of a dye, malachite green, which when complexed with phosphomolybdate at low pH has a high extinction coefficient (at 610 nm). The use of a persulfate oxidation technique to liberate phosphate from lipids increased the simplicity and safety of the method relative to the traditional perchloric acid digestions. The modified method was both accurate (yielding quantitative recoveries of cells added to sediments) and precise (coefficient of variation of less than 5% for cells and sediments). A comparison with an epifluorescence technique indicated that the analysis of lipid-bound phosphate was more rapid and less tedious and could be successfully applied to a wider variety of sediment types. An estimate of the lipid-bound phosphate-to carbon conversion factor based on a diverse enrichment culture from sediments suggested that previous factors for pure cultures may have been too low. PMID- 16348052 TI - Bacterial metabolism of 2,6-xylenol. AB - Strain DM1, a Mycobacterium sp. that utilizes 2,6-xylenol, 2,3,6-trimethylphenol, and o-cresol as sources of carbon and energy, was isolated. Intact cells of Mycobacterium strain DM1 grown with 2,6-xylenol cooxidized 2,4,6-trimethylphenol to 2,4,6-trimethylresorcinol. 4-Chloro-3,5-dimethylphenol prevents 2,6-xylenol from being totally degraded; it was quantitatively converted to 2,6 dimethylhydroquinone by resting cells. 2,6-Dimethylhydroquinone, citraconate, and an unidentified metabolite were detected as products of 2,6-xylenol oxidation in cells that were partially inactivated by EDTA. Under oxygen limitation, 2,6 dimethylhy-droquinone, citraconate, and an unidentified metabolite were released during 2,6-xylenol turnover by resting cells. Cell extracts of 2,6-xylenol-grown cells contained a 2,6-dimethylhydroquinone-converting enzyme. When supplemented with NADH, cell extracts catalyzed the reduction of 2,6-dimethyl-3-hydroxyquinone to 2,6-dimethyl-3-hydroxyhydroquinone. Since a citraconase was also demonstrated in cell extracts, a new metabolic pathway with 2,6-dimethyl-3-hydroxyhydroquinone as the ring fission substrate is proposed. PMID- 16348053 TI - Characterization of Large, Autotrophic Beggiatoa spp. Abundant at Hydrothermal Vents of the Guaymas Basin. AB - Filamentous bacteria, identified as members of the genus Beggiatoa by gliding motility and internal globules of elemental sulfur, occur in massive aggregations at the deep-sea hydrothermal vents of the Guaymas Basin, Gulf of California. Cell aggregates covering the surface of sulfide-emanating sediments and rock chimneys were collected by DS R/V Alvin and subjected to shipboard and laboratory experiments. Each sample collected contained one to three discrete width classes of this organism usually accompanied by a small number of "flexibacteria" (width, 1.5 to 4 mum). The average widths of the Beggiatoa classes were 24 to 32, 40 to 42, and 116 to 122 mum. As indicated by electron microscopy and cell volume/protein ratios, the dominant bacteria are hollow cells, i.e., a thin layer of cytoplasm surrounding a large central liquid vacuole. Activities of Calvin cycle enzymes indicated that at least two of the classes collected possess autotrophic potential. Judging from temperature dependence of enzyme activities and whole-cell CO(2) incorporation, the widest cells were mesophiles. The narrowest Beggiatoa sp. was either moderately thermophilic or mesophilic with unusually thermotolerant enzymes. This was consistent with its occurrence on the flanks of hot smoker chimneys with highly variable exit temperatures. In situ CO(2) fixation rates, sulfide stimulation of incorporation, and autoradiographic studies suggest that these Beggiatoa spp. contribute significantly as lithoautrophic primary producers to the Guaymas Basin vent ecosystems. PMID- 16348054 TI - Rate Equations and Kinetic Parameters of the Reactions Involved in Pyrite Oxidation by Thiobacillus ferrooxidans. AB - Rate equations and kinetic parameters were obtained for various reactions involved in the bacterial oxidation of pyrite. The rate constants were 3.5 muM Fe per min per FeS(2) percent pulp density for the spontaneous pyrite dissolution, 10 muM Fe per min per mM Fe for the indirect leaching with Fe, 90 muM O(2) per min per mg of wet cells per ml for the Thiobacillus ferrooxidans oxidation of washed pyrite, and 250 muM O(2) per min per mg of wet cells per ml for the T. ferrooxidans oxidation of unwashed pyrite. The K(m) values for pyrite concentration were similar and were 1.9, 2.5, and 2.75% pulp density for indirect leaching, washed pyrite oxidation by T. ferrooxidans, and unwashed pyrite oxidation by T. ferrooxidans, respectively. The last reaction was competitively inhibited by increasing concentrations of cells, with a K(i) value of 0.13 mg of wet cells per ml. T. ferrooxidans cells also increased the rate of Fe production from Fe plus pyrite. PMID- 16348056 TI - Influence of Water Potential on gamma-Decalactone Production by the Yeast Sporidiobolus salmonicolor. AB - The influence of water potential on gamma-decalactone production by the yeast Sporidiobolus salmonicolor cultivated in a liquid medium was evaluated by gas chromatographic analysis. Modifications in water potential led to a number of variations in the aroma production. Maximum extracellular production occurred at water activity (a(w)) with a value of 0.99. Further analyses revealed an important phenomenon of cellular accumulation of aroma for a(w) values between 0.97 and 0.99. PMID- 16348055 TI - Nuclear Magnetic Resonance Studies of Poly(3-Hydroxybutyrate) and Polyphosphate Metabolism in Alcaligenes eutrophus. AB - The metabolic pathways of poly(3-hydroxybutyrate) (PHB) and polyphosphate in the microorganism Alcaligenes eutrophus H16 were studied by H, C, and P nuclear magnetic resonance (NMR) spectroscopy and by conventional analytical techniques. A. eutrophus cells accumulated two storage polymers of PHB and polyphosphate in the presence of carbon and phosphate sources under aerobic conditions after exhaustion of nitrogen sources. The solid-state cross-polarization/magic-angle spinning C NMR spectroscopy was used to study the biosynthetic pathways of PHB and other cellular biomass components from C-labeled acetate. The solid-state C NMR analysis of lyophilized intact cells grown on [1-C]acetate indicated that the carbonyl carbon of acetate was selectively incorporated both into the carbonyl and methine carbons of PHB and into the carbonyl carbons of proteins. The P NMR analysis of A. eutrophus cells in suspension showed that the synthesis of intracellular polyphosphate was closely related to the synthesis of PHB. The roles of PHB and polyphosphate in the cells were studied under conditions of carbon, phosphorus, and nitrogen source starvation. Under both aerobic and anaerobic conditions PHB was degraded, whereas little polyphosphate was degraded. The rate of PHB degradation under anaerobic conditions was faster than that under aerobic conditions. Under anaerobic conditions, acetate and 3-hydroxybutyrate were produced as the major extracellular metabolites. The implications of this observation are discussed in connection with the regulation of PHB and polyphosphate metabolism in A. eutrophus. PMID- 16348057 TI - Immunochemical localization of nitrogenase in marine trichodesmium aggregates: relationship to n(2) fixation potential. AB - Colonial aggregation among nonheterocystous filaments of the planktonic marine cyanobacterium Trichodesmium is known to enhance N(2) fixation, mediated by the O(2)-sensitive enzyme complex nitrogenase. Expression of nitrogenase appears linked to the formation of O(2)-depleted microzones within aggregated bacterium associated colonies. While this implies a mechanism by which nonheterocystous N(2) fixation can take place in an oxygenated water column, both the location and regulation of the N(2)-fixing apparatus remain unknown. We used an antinitrogenase polyclonal antibody together with postsection immunocolloidal gold staining and transmission electron microscopy to show that (i) virtually all Trichodesmium cells within a colony possessed nitrogenase, (ii) nitrogenase showed no clear intracellular localization, and (iii) certain associated bacteria contained nitrogenase. Our findings emphasize the critical role coloniality plays in regulating nitrogenase expression in nature. We interpret the potential for a large share of Trichodesmium cells to fix N(2) as an opportunistic response to the dynamic nature of the sea state; during quiescent conditions, aggregation and consequent expression of nitrogenase can proceed rapidly. PMID- 16348058 TI - Response of plant-colonizing pseudomonads to hydrogen peroxide. AB - Colonization of plant root surfaces by Pseudomonas putida may require mechanisms that protect this bacterium against superoxide anion and hydrogen peroxide produced by the root. Catalase and superoxide dismutase may be important in this bacterial defense system. Stationary-phase cells of P. putida were not killed by hydrogen peroxide (H(2)O(2)) at concentrations up to 10 mM, and extracts from these cells possessed three isozymic bands (A, B, and C) of catalase activity in native polyacrylamide gel electrophoresis. Logarithmic-phase cells exposed directly to hydrogen peroxide concentrations above 1 mM were killed. Extracts of logarithmic-phase cells displayed only band A catalase activity. Protection against 5 mM H(2)O(2) was apparent after previous exposure of the logarithmic phase cells to nonlethal concentrations (30 to 300 muM) of H(2)O(2). Extracts of these protected cells possessed enhanced catalase activity of band A and small amounts of bands B and C. A single form of superoxide dismutase and isoforms of catalase were apparent in extracts from a foliar intercellular pathogen, Pseudomonas syringae pv. phaseolicola. The mobilities of these P. syringae enzymes were distinct from those of enzymes in P. putida extracts. PMID- 16348059 TI - Relationship between the Unicellular Red Alga Porphyridium sp. and Its Predator, the Dinoflagellate Gymnodinium sp. AB - Contamination of algae cultivated outdoors by various microorganisms, such as bacteria, fungi, algae, and protozoa, can affect growth and product quality, sometimes causing fast collapse of the cultures. The main contaminant of Porphyridium cultures grown outdoors in Israel is a Gymnodinium sp., a dinoflagellate that feeds on the alga. Comparison of the effects of various environmental conditions, i.e., pH, salinity, and temperature, on Gymnodinium and Porphyridium species revealed that the Gymnodinium sp. has sharp optimum curves, whereas the Porphyridium sp. has a wider range of optimum conditions and is also more resistant to extreme environmental variables. The mode of preying on the alga was observed, and the specificity of the Gymnodinium sp. for the Porphyridium sp. was shown. In addition, Gymnodinium extract was shown to contain enzymatic degrading activity specific to the Porphyridium sp. cell wall polysaccharide. PMID- 16348060 TI - Relationships among Cell Size, Membrane Permeability, and Preservative Resistance in Yeast Species. AB - The rate of uptake of propanoic acid and the cell dimensions were measured for 23 yeasts differing in their resistance to weak-acid-type preservatives. Relationships between reciprocal uptake rate, reciprocal permeability, cell volume, cell area, volume/area, and the MICs of benzoic acid and propanoic acid for the yeasts were tested by correlation analysis on pairs of parameters. The MIC of methylparaben, which is not a weak-acid-type preservative, was included. The most significant relationships found were between both reciprocal uptake rate and reciprocal permeability and the MICs of propanoic and benzoic acids Cell volume, area, and volume/area were each individually correlated with propanoic and benzoic acid MICs, but less strongly. In multiple regression analyses, inclusion of terms for volume, area, or volume/area did not markedly increase the significance. The MIC of methylparaben was unrelated to the uptake and permeability parameters, but did show a correlation with cell volume/area. Schizosaccharomyces pombe was anomalous in having very low permeability. Exclusion of these outlying data revealed particularly strong relationships (P < 0.001) between both reciprocal uptake rate and reciprocal permeability and the benzoic acid MIC. MICs for Zygosaccharomyces bailii isolates were substantially higher than for the other species, and therefore Z. baillii isolates had a large influence on the regressions. However, the relationships observed remained significant even after removal of the Z. bailii data. In showing a correlation between the rate at which propanoic acid enters yeast cells and the ability of the cells to tolerate this and other weak-acid-type preservatives, but not methylparaben, the results suggest that the resistance mechanism, in which preservative is continuously removed from the cell, is a common and major determinant of the preservative tolerance of yeast species. PMID- 16348061 TI - Unaltered Nodulation Competitiveness of a Strain of Bradyrhizobium sp. (Lotus) after a Decade in Soil. AB - A Bradyrhizobium sp. (Lotus) strain that formed a soil population that was highly competitive for nodulation of Lotus pedunculatus 11 years after its introduction into a field soil and a culture of the same strain stored lyophilized were compared with an antibiotic-resistant mutant in respect of their nodulation competitiveness. The mutant was less competitive than the wild-type strain it was isolated from and had to be present at a cell ratio of 5.76:1 in mixed inoculum in sand culture to form 50% of the nodules on L. pedunculatus (50% nodulation value, 5.76). The 50% nodulation values for a soil population of the mutant mixed with soil populations of the lyophilized and field soil strain were, respectively, 6.83 and 5.77, indicating that the field soil strain was not significantly different from the lyophilized strain in nodulation competitiveness. A 50% nodulation value of 11.18 obtained when soil containing a recently established mutant population was mixed with the field soil containing the population established 11 years before, indicating that the plant infection technique underestimated cell numbers of the field soil population by 100%. Nodulation competitiveness was unaffected by the size of the strain populations in the range of 100 to 1,000 cells per g of soil; at 10 cells per g a significant correlation between strain ratios in nodules and in soil was still evident. The results indicated that apparently superior nodulation competitiveness of a well established soil population relative to that of a subsequently introduced strain may not necessarily reflect the intrinsic competitive abilites of the strain(s) involved. The soil strain did not differ from laboratory-maintained cultures in antigenic properties, effectiveness, or whole cell protein electrophoresis profiles. PMID- 16348062 TI - Yeast lipid estimation by enzymatic and nuclear magnetic resonance methods. AB - Low-resolution nuclear magnetic resonance and enzymatic glycerol estimation were compared with a solvent extraction method for estimating the intracellular lipid content of lipid-accumulating yeasts. Both methods correlated well with the solvent extraction procedure and were more convenient with large numbers of samples. PMID- 16348063 TI - High Efficiency of Plating of the Thermophilic Sulfur-Dependent Archaebacterium Sulfolobus acidocaldarius. AB - A procedure for plating Sulfolobus acidocaldarius using Gelrite as the gelling agent is presented. The technique uses a supporting gel of 0.8% (wt/vol) Gelrite and an overlay soft gel of 0.4% (wt/vol) Gelrite, in which the colonies are grown. The plating efficiency was essentially 100%. PMID- 16348064 TI - Comparisons of ruminal fermentation characteristics and microbial populations in bison and cattle. AB - [This corrects the article on p. 2511 in vol. 54.]. PMID- 16348065 TI - Epiphytic Occurrence of Azorhizobium caulinodans and Other Rhizobia on Host and Nonhost Legumes. AB - [This corrects the article on p. 2407 in vol. 55.]. PMID- 16348066 TI - Identification of a Locus Upstream from the Hydrogenase Structural Genes That Is Involved in Hydrogenase Expression in Bradyrhizobium japonicum. AB - A locus involved in the expression of the uptake hydrogenase system of Bradyrhizobium japonicum was identified adjacent to genes encoding the hydrogenase subunits. A cloned fragment of DNA was used to complement to autotrophy a Hup putative regulatory mutant of B. japonicum. The mutant strain lacked hydrogenase activity and synthesized low levels of the large subunit of hydrogenase as determined by Western gels. Tn5-induced mutagenesis located the region within the fragment which was necessary for complementation of the mutant phenotype. The locus identified is adjacent to that encoding the small subunit of hydrogenase; its right border is <0.5 kilobase upstream from the hydrogenase transcriptional start site, and its left border is between 1 and 2.5 kilobases from that start site. However, the locus is outside the region previously shown to contain hup-related genes of B. japonicum. Thus, the localization of this gene describes a previously unidentified hup-related gene on a region of DNA not previously shown to contain hup-specific DNA. PMID- 16348067 TI - Characteristics of woodland rhizobial populations from surface- and deep-soil environments of the sonoran desert. AB - A collection of 74 rhizobial isolates recovered from nodules of the desert woody legumes Prosopis glandulosa, Psorothamnus spinosus, and Acacia constricta were characterized by using 61 nutritional and biochemical tests. We compared isolates from A. constricta and Prosopis glandulosa and tested the hypothesis that the rhizobia from a deep-phreatic rooting zone of a Prosopis woodland in the Sonoran Desert of southern California were phenetically distinct from rhizobia from surface soils. Cluster analysis identified four major homogeneous groups. The first phenon contained slow-growing (SG) Prosopis rhizobia from surface and deep phreatic-soil environments. These isolates grew poorly on most of the media used in the study, probably because of their requirement for a high medium pH. The second group of isolates primarily contained SG Prosopis rhizobia from the deep phreatic rooting environment and included two fast-growing (FG) Psorothamnus rhizobia. These isolates were nutritionally versatile and grew over a broad pH range. The third major phenon was composed mainly of FG Prosopis rhizobia from surface and dry subsurface soils. While these isolates used a restricted range of carbohydrates (including sucrose) as sole carbon sources, they showed better growth on a range of organic acids as sole carbon sources and amino acids as sole carbon and nitrogen sources than did other isolates in the study. They grew better at 36 degrees C than at 26 degrees C. The FG Acacia rhizobia from surface soil environments formed a final major phenon that was distinct from the Prosopis isolates. They produced very high absorbance readings on all of the carbohydrates tested except sucrose, grew poorly on many of the other substrates tested, and preferred a 36 to a 26 degrees C incubation temperature. The surface populations of Prosopis rhizobia required a higher pH for growth and, under the conditions used in this study, were less tolerant of low solute potential and high growth temperature than were phreatic-soil isolates. SG Prosopis rhizobia from phreatic and surface soils were physiologically distinct, suggesting adaptation to their respective soil environments. PMID- 16348068 TI - Profiling and quantitation of bacterial carotenoids by liquid chromatography and photodiode array detection. AB - An analytical method for the profiling and quantitative determination of carotenoids in bacteria is described. Exhaustive extraction of the pigments from four selected bacterial strains required treatment of the cells with potassium hydroxide or liquefied phenol or both before the addition of the extracting solvent (methanol or diethyl ether). The carotenoids in the extracts were separated by nonaqueous reversed-phase liquid chromatography in conjunction with photodiode array absorption detection. The identity of a peak was considered definitive only when both its retention time and absorption spectrum, before and after chemical reactions, matched those of a reference component. In the absence of the latter, most peaks could be tentatively identified. Two examples illustrate how in the analysis of pigmented bacteria errors may result from using nonchromatographic procedures or liquid chromatographic methods lacking sufficient criteria for peak identification. Carotenoids of interest were determined quantitatively when the authentic reference substance was available or, alternatively, were determined semiquantitatively. PMID- 16348069 TI - Action of a fluoranthene-utilizing bacterial community on polycyclic aromatic hydrocarbon components of creosote. AB - Cultures enriched by serial transfer through a mineral salts medium containing fluoranthene were used to establish a stable, seven-member bacterial community from a sandy soil highly contaminated with coal tar creosote. This community exhibited an ability to utilize fluoranthene as the sole carbon source for growth, as demonstrated by increases in protein concentration and changes in absorption spectra when grown on fluoranthene in liquid culture. Biotransformation of other polycyclic aromatic hydrocarbons (PAHs) was verified by demonstrating their disappearance from an artificial PAH mixture by capillary gas chromatography. When grown on fluoranthene as the sole carbon source and subsequently exposed to fluoranthene plus 16 additional PAHs typical of those found in creosote, this community transformed all PAHs present in this defined mixture. After 3 days of incubation, 13 of the original 17 PAH components were degraded to levels below the limit of detection (10 ng/liter). Continued incubation resulted in extensive degradation of the remaining four compounds. The ability of this community to utilize a high-molecular-weight PAH as the sole carbon source, in conjunction with its ability to transform a diverse array of PAHs, suggests that it may be of value in the bioremediation of environments contaminated with PAHs, such as those impacted by creosote. PMID- 16348070 TI - Physiological and Environmental Studies of Sclerotium Formation and Maturation in Isolates of Morchella crassipes. AB - This study provides a set of nutritional and environmental parameters suitable for the growth of morel (Morchella crassipes) sclerotia in the laboratory, using a modification of the jar method of Ower et al. (U.S. patent 4,594,809, June 1986). The optimum nutritional and environmental conditions for morel sclerotium formation and maturation as determined in this study consist of a layer of rye grain supplemented with peptone, yeast extract, trace elements, and Casamino Acids overlaid with perforated aluminum foil and covered with a layer of nutrient poor soil medium in an 8-oz. (ca. 237-ml) glass jar in the dark. We noted that addition of asparagine or aspartic acid as a nitrogen source to the rye also had a beneficial effect on sclerotium formation, while addition of carbon sources had no significant effect. PMID- 16348071 TI - Mechanism of Proteinase Release from Lactococcus lactis subsp. cremoris Wg2. AB - The procedure generally used for the isolation of extracellular, cell-associated proteinases of Lactococcus lactis species is based on the release of the proteinases by repeated incubation and washing of the cells in a Ca-free buffer. For L. lactis subsp. cremoris Wg2, as many as five incubations for 30 min at 29 degrees C are needed in order to liberate 95% of the proteinase. Proteinase release was not affected by chloramphenicol, which indicates that release is not the result of protein synthesis during the incubations. Ca inhibited, while ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) stimulated, proteinase release from the cells. The pH optimum for proteinase release ranged between 6.5 and 7.5, which was higher than the optimum pH of the proteinase measured for casein hydrolysis (i.e., 6.4). Treatment of cells with the serine proteinase inhibitor phenylmethylsulfonyl fluoride prior to the incubations in Ca-free buffer reduced the release of the proteinase by 70 to 80%. The residual proteinase remained cell associated but could be removed by the addition of active L. lactis subsp. cremoris Wg2 proteinase. This suggests that proteinase release from cells of L. lactis subsp. cremoris Wg2 is the result of autoproteolytic activity. From a comparison of the N-terminal amino acid sequence of the released proteinase with the complete amino acid sequence determined from the nucleotide sequence of the proteinase gene, a protein of 180 kilodaltons would be expected. However, a proteinase with a molecular weight of 165,000 was found, which indicated that further hydrolysis had occurred at the C terminus. PMID- 16348072 TI - Induction and Purification of Endo-beta-N-Acetylglucosaminidase from Arthrobacter protophormiae Grown in Ovalbumin. AB - Arthrobacter protophormiae produced a high level of extracellular endo-beta-N acetylglucosaminidase when cells were grown in a medium containing ovalbumin. The enzyme was induced by the glycopeptide fraction of ovalbumin prepared by pronase digestion. Production of the enzyme was also induced by glycoproteins such as yeast invertase and bovine ribonuclease B but not by monosaccharides such as mannose, N-acetylglucosamine, and galactose. The enzyme was purified to homogeneity as demonstrated by polyacrylamide gel electrophoresis and has an apparent molecular weight of about 80,000. The enzyme showed a broad optimum pH in the range of pH 5.0 to 11.0. The enzyme hydrolyzed all heterogeneous ovalbumin glycopeptides, although the hydrolysis rates for hybrid type glycopeptides were very low. The substrate specificity of A. protophormiae endo-beta-N acetylglucosaminidase was very similar to that of Endo-C(II) from Clostridium perfringens. Therefore, the enzyme induction by A. protophormiae seems to have a close relation to the substrate specificity of the enzyme. PMID- 16348073 TI - High-Frequency Transformation, by Electroporation, of Lactococcus lactis subsp. cremoris Grown with Glycine in Osmotically Stabilized Media. AB - An efficient method for genetic transformation of lactococci by electroporation is presented. Highly competent lactococci for electrotransformation were obtained by growing cells in media containing high concentrations of glycine and 0.5 M sucrose as the osmotic stabilizers. These cells could be stored at -85 degrees C without loss of competence. With Lactococcus lactis subsp. cremoris BC101, a transformation frequency of 5.7 x 10 transformants per mug of pIL253 DNA was obtained, which represents 5% of the surviving cells. All the lactococcal strains tested could be transformed by the present method. PMID- 16348074 TI - Organization of Genes Required for the Oxidation of Methanol to Formaldehyde in Three Type II Methylotrophs. AB - Restriction maps of genes required for the synthesis of active methanol dehydrogenase in Methylobacterium organophilum XX and Methylobacterium sp. strain AM1 have been completed and compared. In these two species of pink-pigmented, type II methylotrophs, 15 genes were identified that were required for the expression of methanol dehydrogenase activity. None of these genes were required for the synthesis of the prosthetic group of methanol dehydrogenase, pyrroloquinoline quinone. The structural gene required for the synthesis of cytochrome c(L), an electron acceptor uniquely required for methanol dehydrogenase, and the genes encoding small basic peptides that copurified with methanol dehydrogenases were closely linked to the methanol dehydrogenase structural genes. A cloned 22-kilobase DNA insert from Methylsporovibrio methanica 81Z, an obligate type II methanotroph, complemented mutants that contained lesions in four genes closely linked to the methanol dehydrogenase structural genes. The methanol dehydrogenase and cytochrome c(L) structural genes were found to be transcribed independently in M. organophilum XX. Only two of the genes required for methanol dehydrogenase synthesis in this bacterium were found to be cotranscribed. PMID- 16348075 TI - Fermentation of Cellulosic Substrates in Batch and Continuous Culture by Clostridium thermocellum. AB - Fermentation of dilute-acid-pretreated mixed hardwood and Avicel by Clostridium thermocellum was compared in batch and continuous cultures. Maximum specific growth rates per hour obtained on cellulosic substrates were 0.1 in batch culture and >0.13 in continuous culture. Cell yields (grams of cells per gram of substrate) in batch culture were 0.17 for pretreated wood and 0.15 for Avicel. Ethanol and acetate were the main products observed under all conditions. Ethanol:acetate ratios (in grams) were approximately 1.8:1 in batch culture and generally slightly less than 1:1 in continuous culture. Utilization of cellulosic substrates was essentially complete in batch culture. A prolonged lag phase was initially observed in batch culture on pretreated wood; the length of the lag phase could be shortened by addition of cell-free spent medium. In continuous culture with approximately 5 g of glucose equivalent per liter in the feed, substrate conversion relative to theoretical ranged from 0.86 at a dilution rate (D) of 0.05/h to 0.48 at a D of 0.167/h for Avicel and from 0.75 at a D of 0.05/h to 0.43 at a D of 0.11/h for pretreated wood. At feed concentrations of <4.5 g of glucose equivalent per liter, conversion of pretreated wood was 80 to 90% at D = 0.083/h. Lower conversion was obtained at higher feed substrate concentrations, consistent with a limiting factor other than cellulose. Free Avicelase activities of 12 to 84 mU/ml were observed, with activity increasing in this order: batch cellobiose, batch pretreated wood < batch Avicel, continuous pretreated wood < continuous Avicel. Free cellulase activity was higher at increasing extents of substrate utilization for both pretreated wood and Avicel under all conditions tested. The results indicate that fermentation parameters, with the exception of free cellulase activity, are essentially the same for pretreated mixed hardwood and Avicel under a variety of conditions. Hydrolysis yields obtained with C. thermocellum cellulase acting either in vitro or in vivo were comparable to those previously reported for Trichoderma reesei on the same substrates. PMID- 16348076 TI - Adsorption-desorption characteristics of polychlorinated biphenyls on various polymers commonly found in laboratories. AB - Adsorption and desorption of Aroclor 1254 (a mixture of polychlorinated biphenyls [PCBs]) from 0.25% (wt/vol) aqueous solutions of Triton X-100 on polymeric materials common in laboratories (red vacuum rubber, latex, Tygon, Norprene, manosil, polyethylene, polypropylene, phenoxy resin, nylon, and Teflon) is described. Teflon, nylon, and phenoxy resins showed the lowest adsorptions, with efficiencies of 3.4, 22.9, and 33.0%, respectively. The other polymers adsorbed more than 90% of Aroclor 1254 under similar conditions. Adsorption of PCBs was found to increase with the lipophilicity of the polymer and to be irreversible on soft polymers. The variation in the adsorption efficiencies of these polymers toward PCBs is apparently related to variations in the chemical composition, electronic properties, and degree of softness of these polymers. The present study showed that Teflon, with less than 4% adsorption and more than 99% desorption of Aroclor 1254, is the best candidate for use in biological laboratory work. PMID- 16348077 TI - Cloning and Expression of a Schwanniomyces occidentalis alpha-Amylase Gene in Saccharomyces cerevisiae. AB - An alpha-amylase gene (AMY) was cloned from Schwanniomyces occidentalis CCRC 21164 into Saccharomyces cerevisiae AH22 by inserting Sau3AI-generated DNA fragments into the BamHI site of YEp16. The 5-kilobase insert was shown to direct the synthesis of alpha-amylase. After subclones containing various lengths of restricted fragments were screened, a 3.4-kilobase fragment of the donor strain DNA was found to be sufficient for alpha-amylase synthesis. The concentration of alpha-amylase in culture broth produced by the S. cerevisiae transformants was about 1.5 times higher than that of the gene donor strain. The secreted alpha amylase was shown to be indistinguishable from that of Schwanniomyces occidentalis on the basis of molecular weight and enzyme properties. PMID- 16348078 TI - Beneficial Effects of Nickel on Pseudomonas saccharophila under Nitrogen-Limited Chemolithotrophic Conditions. AB - Addition of nickel stimulated growth and nitrogenase activity of Pseudomonas saccharophila under nitrogen-limited chemolithotrophic conditions, apparently because of a significant increase in expression of uptake hydrogenase activity. Inhibition of hydrogenase expression by 50 muM EDTA was relieved by nickel over a wide concentration range (1 to 200 muM). Co, Zn, Mn, and Cu stimulated expression of hydrogenase activity, but to a much lesser degree than nickel, and Fe, Mg, SeO(4), and SeO(3) did not increase expression. Nickel in individual combination with Mg, Fe, SeO(3), and SeO(4) resulted in activities that were essentially the same as that with nickel alone. Hydrogenase synthesis required the presence of nickel, and repression by O(2) was alleviated by increasing the concentration of added nickel. Cells placed under hydrogenase derepression conditions showed progressive incorporation of radioactive nickel to a much greater extent than did cells which were not derepressed. PMID- 16348079 TI - Host Restriction and Transduction in Rhizobium meliloti. AB - A host restriction difference exists between Rhizobium meliloti Rm41 and SU47 exists as indicated by the reduce plating efficiency of transducing phage PhiM12h1. Restriction can be attenuated by incubating cells at 42 degrees C for 3 h; this procedure overcomes a block to transduction from SU47 to Rm41. PMID- 16348080 TI - Requirement for a Microbial Consortium To Completely Oxidize Glucose in Fe(III) Reducing Sediments. AB - In various sediments in which Fe(III) reduction was the terminal electron accepting process, [C]glucose was fermented to C-fatty acids in a manner similar to that observed in methanogenic sediments. These results are consistent with the hypothesis that, in Fe(III)-reducing sediments, fermentable substrates are oxidized to carbon dioxide by the combined activity of fermentative bacteria and fatty acid-oxidizing, Fe(III)-reducing bacteria. PMID- 16348081 TI - Simple Method To Detect beta-Galactosidase. AB - A simple and rapid method was developed to detect beta-galactosidase by using alpha- or beta-naphthyl-beta-d-galactopyranoside as substrate and fast garnet GBC as a dye coupler following polyacrylamide gel electrophoresis. This method was specific for beta-galactosidase but not for phospho-beta-galactosidase. PMID- 16348082 TI - Development of an Adhesion Assay and Characterization of an Adhesion-Deficient Mutant of Pseudomonas fluorescens. AB - A sand column adhesion assay was developed which distinguishes the adhesion abilities of a number of pseudomonads isolated from fine sandy loam. Pseudomonas fluorescens Pf0-1 which adhered at >90% of the total cells added was subjected to transposon Tn5 insertion mutagenesis. From 2,500 Pf0-1::Tn5 mutants examined in the sand column assay, two adhesion-deficient Pf0-1 mutants showing <50% attachment were isolated. Marker exchange analysis of one of these mutants, Pf0 5, confirmed that the decreased adhesion was linked to the Tn5 insertion in the chromosome. The growth rate of Pf0-5 in enriched media and sterile soil was similar to that of the wild type; in minimal medium, however, Pf0-5 grew faster. In a soil column assay, less Pf0-5 than wild-type bacteria were recovered, suggesting a decreased ability to persist in soil. A 34-kilodalton major outer membrane protein present in the wild type was missing in Pf0-5. Transmission electron microscopy of the cell surface revealed that the wild-type possessed polar flagella which were absent in the mutant. PMID- 16348083 TI - Intermediary Metabolite Concentrations in Xylulose- and Glucose-Fermenting Saccharomyces cerevisiae Cells. AB - Glucose and xylulose fermentation and product formation by Saccharomyces cerevisiae were compared in batch culture under anaerobic conditions. In both cases the main product was ethanol, with glycerol, xylitol, and arabitol produced as by-products. During glucose and xylulose fermentation, 0.74 and 0.37 g of cell mass liter, respectively, were formed. In glucose-fermenting cells, the carbon balance could be closed, whereas in xylulose-fermenting cells, about 25% of the consumed sugar carbon could not be accounted for. The rate of sugar consumption was 3.94 mmol g of initial biomass h for glucose and 0.39 mmol g of initial biomass h for xylulose. Concentrations of the intermediary metabolites fructose 1,6-diphosphate (FDP), pyruvate (PYR), sedoheptulose 7-phosphate (S7P), erytrose 4-phosphate, citrate (CIT), fumarate, and malate were compared for both types of cells. Levels of FDP, PYR, and CIT were lower, and levels of S7P were higher in xylulose-fermenting cells. After normalization to the carbon consumption rate, the levels of FDP were approximately the same, whereas there was a significant accumulation of S7P, PYR, CIT, and malate, especially of S7P, in xylulose fermenting cells compared with in glucose-fermenting cells. In the presence of 15 muM iodoacetate, an inhibitor of the enzyme glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12), FDP levels increased and S7P levels decreased in xylulose-assimilating cells compared with in the absence of the inhibitor, whereas fermentation was slightly slowed down. The specific activity of transaldolase (EC 2.2.1.2), the pentose phosphate pathway enzyme reacting with S7P and glyceraldehyde-3-phosphate, was essentially the same for both glucose- and xylulose-fermenting cells. It was, however, several orders of magnitude lower than that reported for a Torula yeast and Candida utilis. The presence of iodoacetate did not influence the activity of transaldolase in xylulose fermenting cells. The results are discussed in terms of a competition between the pentose phosphate pathway and glycolysis for the common metabolite, glyceraldehyde-3-phosphate, which would explain the low rates of xylulose assimilation and ethanol production from xylulose by S. cerevisiae. PMID- 16348084 TI - Esterase Activities of Fibrobacter succinogenes subsp. succinogenes S85. AB - Cells of the anaerobic ruminal bacterium Fibrobacter succinogenes subsp. succinogenes S85 (formerly Bacteroides succinogenes) exhibit arylesterase activity. When cells were grown on cellulose, it was found that 69% of the total esterase activity was extracellular while 65% was nonsedimentable upon centrifugation of the culture supernatant at 100,000 x g. Disruption of the cells by various different methods failed to increase the esterase activity, indicating that the substrate was fully accessible to esterase enzymes in intact cells. During growth of cells with either glucose or cellulose as the sole carbon source, the increase in acetylesterase activity corresponded to an increase in cell density, suggesting constitutive production. The enzyme(s) hydrolyzed alpha naphthyl, p-nitrophenyl, and 4-methylumbelliferyl derivatives of acetic acid; xylose tetraacetate; glucose pentaacetate; acetylxylan; and a polymer composed of ferulic acid, arabinose, and xylose in molar proportions of 1:1.1:2.2 (FAX). These data demonstrate the presence of an acetylxylan esterase and a ferulic acid esterase. The cleavage of FAX also documents the presence of an alpha-l arabinofuranosidase. PMID- 16348085 TI - Plasmid-Determined Copper Resistance in Pseudomonas syringae from Impatiens. AB - A strain of Pseudomonas syringae was recently identified as the cause of a new foliar blight of impatiens. The bacterium was resistant to copper compounds, which are used on a variety of crops for bacterial and fungal disease control. The bacterium contained a single 47-kilobase plasmid (pPSI1) that showed homology to a copper resistance operon previously cloned and characterized from P. syringae pv. tomato plasmid pPT23D (D. Cooksey, Appl. Environ. Microbiol. 53:454 456, 1987). pPSI1 was transformed by electroporation into a copper-sensitive P. syringae strain, and the resulting transformants were copper resistant. A physical map of pPSI1 was constructed, and the extent of homology to pPT23D outside the copper resistance operon was determined in Southern hybridizations. The two plasmids shared approximately 20 kilobases of homologous DNA, with the remainder of each plasmid showing no detectable homology. The homologous regions hybridized strongly, but there was little or no conservation of restriction enzyme recognition sites. PMID- 16348086 TI - Biodegradation of Dinoseb (2-sec-Butyl-4,6-Dinitrophenol) in Several Idaho Soils with Various Dinoseb Exposure Histories. AB - We examined the ability of native microorganisms in various Idaho soils to degrade dinoseb and studied some physical and chemical soil characteristics which might affect the biodegradation process. Dinoseb biodegradation rates were higher in silt-loam soils than in loamy-sand soils. Biodegradation rates were not influenced by previous exposure of the soils to dinoseb. Bacterial numbers, measured by standard plate counts on soil extract agar, were the best predictors of biodegradation rates, accounting for 53% of the variability between soils. Soil nitrate-N inhibited dinoseb biodegradation and accounted for 39% of the variability. Sorption of dinoseb to soil surfaces also appeared to influence biodegradation rates. No other soil parameter contributed significantly to the variability in biodegradation rates. Persistence of dinoseb in one soil was due to inhibition of biodegradation by nitrate, while in another soil persistence appeared to be due to lack of native degradative microorganisms. PMID- 16348087 TI - Fungal catabolism of crown gall opines. AB - This study was conducted to determine the capacities of 37 fungi to utilize various crown gall opines as their sole carbon and nitrogen source. One strain of Fusarium solani, two of Cylindrocarpon destructans, and six of Cylindrocarpon heteronema catabolized octopine, mannopine, octopinic acid, succinamopine, or a combination of these opines. One C. heteronema and one Fusarium dimerum strain grew only on succinamopine. None of the fungal isolates had the ability to grow on nopaline. The catabolism of opines by fungi was confirmed by the disappearance of the opine from the growth medium and by an increase in final mycelial dry weight with rising initial concentration of test substrate. This study thus shows that the catabolism of opines is not restricted to bacteria. PMID- 16348088 TI - Sulfur-containing amino acids as precursors of thiols in anoxic coastal sediments. AB - Sulfur-containing amino acids were examined as precursors for thiols in anoxic coastal sediments. Substrates (10 to 100 muM) were anaerobically incubated with sediment slurries; thiols were assayed as isoindole derivatives by high performance liquid chromatography; and microbial transformations of thiols, in contrast to their chemical binding by sediment particles, were identified by inhibition with a mixture of chloramphenicol and tetracycline. Methionine and homocysteine were transformed to methanethiol and 3-mercaptopropionate (3-MPA); methionine stimulated mainly methanethiol production, whereas homocysteine generated more 3-MPA than methanethiol. 2-Keto-4-methiolbutyrate yielded results similar to those with methionine, indicating that demethiolation yields methanethiol at the keto-acid level. Glutathione gave rise to cysteine, which was further transformed to 3-mercaptopyruvate and thence to mercaptoacetate and mercaptoethanol. Mercaptoethanol was oxidized to mercaptoacetate, which was biologically consumed. In conclusion, sulfurcontaining amino acids contribute to the range of thiols that occur in anoxic coastal sediments. New metabolic and environmental transformations were identified: the production of 3-MPA as a metabolite of methionine and the transformation of mercaptopyruvate to mercaptoethanol and mercaptoacetate. PMID- 16348089 TI - Characterization of pXV10A, a Copper Resistance Plasmid in Xanthomonas campestris pv. vesicatoria. AB - The efficacy of copper bactericides for control of Xanthomonas campestris pv. vesicatoria in eastern Oklahoma tomato fields was evaluated. Copper bactericides did not provide adequate control, and copper-resistant (Cu) strains of the pathogen were isolated. The Cu genes in these strains were located on a large indigenous plasmid designated pXV10A. The host range of pXV10A was investigated; this plasmid was efficiently transferred into 8 of 11 X. campestris pathovars. However, the transfer of pXV10A to other phytopathogenic genera was not detected. DNA hybridization experiments were performed to characterize the Cu genes on pXV10A. A probe containing subcloned Cu genes from X. campestris pv. vesicatoria E3C5 hybridized to pXV10A; however, a subclone containing Cu genes from P. syringae pv. tomato PT23 failed to hybridize to pXV10A. Further DNA hybridization experiments were performed to compare pXV10A with pXvCu plasmids, a heterogenous group of Cu plasmids present in strains of X. campestris pv. vesicatoria from Florida. These studies indicated that the Cu genes on pXV10A and pXvCu plasmids share nucleotide sequence homology and may have a common origin. Further experiments showed that these plasmids are distinctly different because pXV10A did not contain sequences homologous to IS476, an insertion sequence present on pXvCu plasmids. PMID- 16348090 TI - Odorous Sulfur Compounds Emitted during Production of Compost Used as a Substrate in Mushroom Cultivation. AB - Large-scale composting facilities are known to cause environmental problems, mainly through pungent air emitted by composting material. In air samples taken above stacks set up to prepare compost used as a substrate in mushroom cultivation, several volatile compounds were identified by means of the coupled techniques of gas chromatography and mass spectrography. Among the compounds identified, sulfur-containing compounds [H(2)S, COS, CH(3)SH, CS(2), (CH(3))(2)S, (CH(3))(2)S(2), and (CH(3))(2)S(3)] are the most conspicuous in causing a nuisance. Quantification of these compounds was performed by concentrating a relatively small air sample on Tenax GC. The sampling method appeared to be very useful under field conditions. During the composting process, the concentration of the volatile sulfur compounds in emitted air ranged from 1 to 35 mumol/m. The highest concentrations were obtained at the end of the outdoor process. Total sulfur emission amounted to 8.3 mg of sulfur per kg (fresh weight) of compost. The end product still contained 2.58 g of sulfur per kg (fresh weight) of compost. Suggestions about the origin of the volatile sulfur compounds are made. PMID- 16348091 TI - pAMbeta1-Associated Mobilization of Proteinase Plasmids from Lactococcus lactis subsp. lactis UC317 and L. lactis subsp. cremoris UC205. AB - A combination of plasmid curing and DNA-DNA hybridization data facilitated the identification of proteinase plasmids of 75 (pCI301) and 35 kilobases (pCI203) in the multi-plasmid-containing strains Lactococcus lactis subsp. lactis UC317 and L. lactis subsp. cremoris UC205, respectively. Both plasmids were transferred by conjugation to a plasmid-free background only after introduction of the conjugative streptococcal plasmid, pAMbeta1. All Prt transconjugants from matings involving either donor contained enlarged recombinant Prt plasmids. UC317-derived transconjugants were separable into different classes based on the presence of differently sized cointegrate plasmids and on segregation of the pCI301-derived Lac and Prt markers. All UC205-derived transconjugants harbored a single enlarged plasmid that was a cointegrate between pCI203 and pAMbeta1. The identification of prt genes on pCI301 and pCI203 derivatives was achieved by a combination of restriction enzyme and hybridization analyses. PMID- 16348092 TI - Identification of the Minimal Replicon of Lactococcus lactis subsp. lactis UC317 Plasmid pCI305. AB - Replication functions of the stable, cryptic 8.7-kilobase (kb) plasmid pCI305 from multi-plasmid-containing Lactococcus lactis subsp. lactis UC317 were studied. Analysis of this replicon was facilitated by the construction of replication probe vectors that consisted of the pBR322 replication region, a pUC18-derived multiple cloning site, and either the cat gene of pC194 (pCI341; 3.1 kb) or the erm gene of pAMbeta1 (pCI3330; 4.0 kb). Plasmid pCI305 was introduced into plasmid-free L. lactis subsp. lactis MG1363Sm, a streptomycin resistant derivative of MG1363, by a transformation procedure with the 75-kb lactose-proteinase plasmid pCI301 of UC317 as a marker plasmid. A combination of transposon Tn5 mutagenesis and subcloning in pCI341 and pCI3330 with individual Tn5 insertions around the replication region facilitated the identification of a 1.6-kb minimal replicon on pCI305. This region was separable into two domains: (i) a 1.3-kb region (repB) encoding a trans-acting function (in vitro transcription-translation studies suggested the involvement of a 48-kilodalton protein); and (ii) a 0.3-kb region (repA) sufficient to direct replication when provided with repB in trans and thus probably containing the origin of replication. Lactococcus-Escherichia coli shuttle vectors based on the pCI305 replication region were constructed. PMID- 16348093 TI - Mn(II) Regulation of Lignin Peroxidases and Manganese-Dependent Peroxidases from Lignin-Degrading White Rot Fungi. AB - Two families of peroxidases-lignin peroxidase (LiP) and manganese-dependent lignin peroxidase (MnP)-are formed by the lignin-degrading white rot basidiomycete Phanerochaete chrysosporium and other white rot fungi. Isoenzymes of these enzyme families carry out reactions important to the biodegradation of lignin. This research investigated the regulation of LiP and MnP production by Mn(II). In liquid culture, LiP titers varied as an inverse function of and MnP titers varied as a direct function of the Mn(II) concentration. The extracellular isoenzyme profiles differed radically at low and high Mn(II) levels, whereas other fermentation parameters, including extracellular protein concentrations, the glucose consumption rate, and the accumulation of cell dry weight, did not change significantly with the Mn(II) concentration. In the absence of Mn(II), extracellular LiP isoenzymes predominated, whereas in the presence of Mn(II), MnP isoenzymes were dominant. The release of CO(2) from C-labeled dehydrogenative polymerizate lignin was likewise affected by Mn(II). The rate of CO(2) release increased at low Mn(II) and decreased at high Mn(II) concentrations. This regulatory effect of Mn(II) occurred with five strains of P. chrysosporium, two other species of Phanerochaete, three species of Phlebia, Lentinula edodes, and Phellinus pini. PMID- 16348094 TI - Exoprotease Activity of Two Marine Bacteria during Starvation. AB - Exoprotease activity during 120 h of total energy and nutrient starvation was examined in two marine bacteria, Vibrio sp. strain S14 and Pseudomonas sp. strain S9. The activity was determined by spectrophotometric measurement of the rate of release of soluble color from an insoluble azure dye derivative of hide powder (hide powder azure). Starved cells of both strains (5 h for S14, and 4 or 24 h for S9) showed greater extracellular proteolytic activity than at the onset of starvation. The exoprotease activity of cells starved for longer periods of time then decreased, but was found to be present at significant levels throughout the starvation period studied (120 h). The accumulation of exoprotease activity in the bulk phase during starvation indicated that both strains constitutively excreted extracellular proteases. As deduced from experiments with chloramphenicol, de novo protein synthesis during starvation was required for the production and/or release of the exoproteases into the surrounding environment. The degradation of hide powder azure allowed an immediate increase in respiration rate, also by long-term-starved cells. This suggests that metabolic systems are primed to respond to the availability of substrates, allowing the cells to recover rapidly. The regulation of exoprotease activity was also studied and found to be different in the two strains. Casamino Acids repressed exoprotease activity in Pseudomonas sp. strain S9, whereas a mechanism similar to catabolite repression was found for Vibrio sp. strain S14 in that glucose repressed activity and cyclic AMP reversed this effect. The exoproteases appeared to be metalloproteinases because the addition of EDTA to cell-free starvation supernatants from both strains significantly inhibited the activity of the proteases. PMID- 16348095 TI - Symbiotic effectiveness of indigenous soybean bradyrhizobia as related to serological, morphological, rhizobitoxine, and hydrogenase phenotypes. AB - A Collection of 360 isolates of Bradyrhizobium japonicum was developed from soybean (Glycine max [L.] Merrill) nodules taken from 18 locations in Delaware. The isolates were characterized serologically with an enzyme-linked immunosorbent assay, morphologically by colony type on yeast extract-mannitol agar, and for production of rhizobitoxine symptoms with soybean plants. These analyses revealed 12 and 3 groups based on serology and morphology, respectively. The more common identifiable isolates were in serogroups 94, 6, 122, and 76. Nearly 33% of the isolates were rated nonreactive with all of the antisera tested. Overall, 18% of the isolates produced rhizobitoxine symptoms, and these were associated with five known serogroups (31, 46, 76, 94, and 130) and the nonreactive grouping, but with only one colony type. A subsample of 92 isolates was rated for N(2)-fixing ability in the greenhouse and for hydrogenase phenotype in the laboratory. The nitrogen content of plant shoots was strongly and comparably related to both the serological and morphological groupings. Rhizobitoxine and hydrogenase phenotypes were relatively poor predictors of symbiotic effectiveness. Among the serologically reactive isolates, those in serogroups 38-115, 122, and 110 fixed the most N(2), whereas one colony type (that containing isolates producing rhizobitoxine) was clearly inferior to the remaining two morphological groupings. Isolates displaying hydrogenase activity (approximately 15% of the isolates tested) correlated with three serologically reactive groupings (serogroups 110 and 122 and a 122/123 cross-reactive group) and two colony types, none of which coincided with groupings containing bradyrhizobia rated positive for rhizobitoxine production. PMID- 16348096 TI - Efficiencies of recovery of bdellovibrios from brackish- water environments by using various bacterial species as prey. AB - A total of 44 bacterial species subdivided into 10 trial experiments have been used as prey for the recovery of bdellovibrios from samples of water from a brackish tidal pond and an aquarium saltwater tank. In an initial investigation, the recovery efficiency of each of the test bacterial species was compared with that of a designated standard prey, Vibrio parahaemolyticus P-5. The results revealed that in each case strain P-5 yielded an equal or significantly greater number of plaques of bdellovibrios than the test prey with but a single exception, strain CS5. In repeat experiments, CS5 yielded fewer plaques than P-5. To determine whether the use of multiple bacterial species compared with a single species as prey would increase the number of PFU of bdellovibrios recovered, material from plaques appearing on each of the test prey in the respective trials was sequentially subcultured onto two respective agar plates, the first containing as prey V. parahaemolyticus P-5 and the second containing the initial test organism. In nearly every case, subculture of plaques from lawns of the test prey to P-5 resulted in plaque formation. On the basis of the results, the use of several test prey and P-5 did not result in the recovery of any more bdellovibrio PFU than the use of P-5 alone. In this study, V. parahaemolyticus P-5 was observed to be the most efficient prey for the recovery of bdellovibrios from moderate salt water. PMID- 16348097 TI - Effects of Acid stress on aerobic decomposition of algal and aquatic macrophyte detritus: direct comparison in a radiocarbon assay. AB - Radiolabeled phytoplankton and macrophyte lignocelluloses were incubated at pHs 4 and 7 in water from a naturally acidic freshwater wetland (Okefenokee Swamp; ambient pH, 3.8 to 4.2), a freshwater reservoir (L-Lake; pH 6.7 to 7.2), and a marine marsh (Sapelo Island; pH approximately 7.8). The data suggest that acidity is an important factor in explaining the lower decomposition rates of algae in Okefenokee Swamp water relative to L-Lake or Sapelo Island water. The decomposition of algal substrate was less sensitive to low pH ( approximately 5 to 35% inhibition) than was the decomposition of lignocellulose ( approximately 30 to 70% inhibition). These substrate-dependent differences were greater and more consistent in salt marsh than in L-lake incubations. In both freshwater sites, the extent to which decomposition was suppressed by acidity was greater for green algal substrate than for mixed diatom or blue-green algal (cyanobacteria) substrates. The use of different bases to adjust pH or incubation in a defined saltwater medium had no significant effect on substrate-dependent differences. Although pH differences with lignocellulose were larger in marine incubations, amendment of lakewater with marine bacteria or with calcium, known to stabilize exoenzymes in soils, did not magnify the sensitivity of decomposition to acid stress. PMID- 16348098 TI - Oxidation of Lignin-Related Aromatic Alcohols by Cell Suspensions of Methylosinus trichosporium. AB - Cell suspensions of Methylosinus trichosporium oxidized the aromatic alcohols benzyl alcohol, vanillyl alcohol, and veratryl alcohol to the corresponding aldehydes, and with the exception of vanillyl alcohol, the aldehydes were further oxidized to the corresponding aromatic acids. No other transformation was observed, and the methoxyl moieties attached to the aromatic nucleus remained intact. More than 70% of the alcohol oxidized could be accounted for by aldehyde and/or acid. Investigation of the inhibitor kinetics of EDTA or p nitrophenylhydrazine (specific for NAD-independent methanol dehydrogenase in methylotrophs) on aromatic alcohol oxidation revealed noncompetitive inhibition in which the V(max) was decreased but the K(m) remained unchanged. The pattern of inhibition of aromatic alcohol oxidation matched that of methanol oxidation, and the K(m) values for all of the substrates were similar (12 to 16 mM). The results indicate that the initial step in the oxidation of aromatic alcohols was similar to that for methanol, and because oxidation was incomplete (i.e., only the corresponding aldehyde or acid was produced), there may be some biotechnological advantages in using whole cells of methylotrophs to facilitate aromatic biotransformations. PMID- 16348099 TI - Mannitol Metabolism in Lentinus edodes, the Shiitake Mushroom. AB - Mannitol metabolism was evaluated in fruiting bodies of Lentinus edodes. Cell extracts were prepared from fruiting bodies, and key enzymes involved in mannitol metabolism were assayed, including hexokinase, mannitol dehydrogenase, mannitol-1 phosphate dehydrogenase, mannitol-1-phosphatase, and fructose-6-phosphatase. Mannitol dehydrogenase, fructose-6-phosphatase, mannitol-1-phosphatase, and hexokinase activities were found in extracts of fruiting bodies. However, mannitol-1-phosphate dehydrogenase activity was not detected. Mycelial cultures were grown in an enriched liquid medium, and enzymes of the mannitol cycle were assayed in cell extracts of rapidly growing cells. Mannitol-1-phosphate dehydrogenase activity was also not found in mycelial extracts. Hence, evidence for a complete mannitol cycle both in vegetative mycelia and during mushroom development was lacking. The pathway of mannitol synthesis in L. edodes appears to utilize fructose as an intermediate. PMID- 16348100 TI - Rubber-degrading enzyme from a bacterial culture. AB - Rubber-degrading activity was found in the extracellular culture medium of Xanthomonas sp. strain 35Y which was grown on natural rubber latex. Natural rubber in the latex state was degraded by the crude enzyme, and two fractions were separately observed by gel permeation chromatography of the reaction products. One fraction was of higher molecular weight (HMW) with a very wide MW distribution from 10 to 10, and the other fraction was of lower molecular weight (LMW) with a MW of a few hundred. H-nuclear magnetic resonance spectra of the partially purified fractions were those expected of cis-1,4-polyisoprene mixtures with the structure OHC-CH(2)-(-CH(2)-C(-CH(3)) = CH-CH(2)-)(n)-CH(2)-C(=O)-CH(3), with average values of n of about 113 and 2 for HMW and LMW fractions, respectively. The LMW fraction consisted mostly of one component in gas-liquid chromatography as well as in gel permeation chromatography, and the main component was identified as 12-oxo-4,8-dimethyl trideca-4,8-diene-1-al (acetonyl diprenyl acetoaldehyde, A(l)P(2)A(t)) by C-nuclear magnetic resonance and gas chromatography-mass spectra. Not only the latices of natural and synthetic isoprene rubber, but also some kinds of low-MW polyisoprene compounds of cis-1,4 type, were degraded by the crude enzyme. The rubber-degrading reaction was found to be at least partly oxygenase catalyzed from the incorporation of O into A(l)P(2)A(t) under an O(2) atmosphere. PMID- 16348101 TI - Transient-state analysis of metabolic fluxes in crabtree-positive and crabtree negative yeasts. AB - In bakers' yeast, an immediate alcoholic fermentation begins when a glucose pulse is added to glucose-limited, aerobically grown cells. The mechanism of this short term Crabtree effect was investigated via a comparative enzymic analysis of eight yeast species. It was established that the fermentation rate of the organisms upon transition from glucose limitation to glucose excess is positively correlated with the level of pyruvate decarboxylase (EC 4.1.1.1). In the Crabtree negative yeasts, the pyruvate decarboxylase activity was low and did not increase when excess glucose was added. In contrast, in the Crabtree-positive yeasts, the activity of this enzyme was on the average sixfold higher and increased after exposure to glucose excess. In Crabtree-negative species, relatively high activities of acetaldehyde dehydrogenases (EC 1.2.1.4 and EC 1.2.1.5) and acetyl coenzyme A synthetase (EC 6.2.1.1), in addition to low pyruvate decarboxylase activities, were present. Thus, in these yeasts, acetaldehyde can be effectively oxidized via a bypass that circumvents the reduction of acetaldehyde to ethanol. Growth rates of most Crabtree-positive yeasts did not increase upon transition from glucose limitation to glucose excess. In contrast, the Crabtree-negative yeasts exhibited enhanced rates of biomass production which in most cases could be ascribed to the intracellular accumulation of reserve carbohydrates. Generally, the glucose consumption rate after a glucose pulse was higher in the Crabtree-positive yeasts than in the Crabtree-negative yeasts. However, the respiratory capacities of steady-state cultures of Crabtree-positive yeasts were not significantly different from those of Crabtree-negative yeasts. Thus, a limited respiratory capacity is not the primary cause of the Crabtree effect in yeasts. Instead, the difference between Crabtree-positive and Crabtree-negative yeasts is attributed to differences in the kinetics of glucose uptake, synthesis of reserve carbohydrates, and pyruvate metabolism. PMID- 16348102 TI - Initial test of the benchmark chemical approach for predicting microbial transformation rates in aquatic environments. AB - Using 2,4-dichlorophenoxyacetic acid methyl ester (2,4-DME) as a benchmark chemical, we determined relative pseudo-first-order rate coefficients for the butoxyethyl ester of 2,4-dichlorophenoxyacetic acid (2,4-DBE), methyl parathion, and methyl-3-chlorobenzoate in a diversity of microbial samples, including water, sediment, biofilm, and floating microbial mats collected from a laboratory mesocosm as well as from streams, lakes, and wetlands in Georgia and Florida. The decreasing order of reactivity for relative microbial transformation rates was 2,4-DBE > 2,4-DME > methyl-3-chlorobenzoate > methyl parathion. Half-lives of the chemicals varied about 60-fold depending on the chemical and microbial sample. Relative rate coefficients, however, typically varied only about threefold for field-collected samples. Relative rate coefficients determined with samples from a laboratory mesocosm were consistently low compared with the field sample data. Overall, the data indicated that microbial transformation rates of a chemical can be satisfactorily inferred for a wide variety of microbial habitats-such as water, biofilm, or a sediment-on the basis of its transformation rate relative to that of an appropriate benchmark chemical by using a single type of microbial sample. PMID- 16348103 TI - Reductive dehalogenation of a nitrogen heterocyclic herbicide in anoxic aquifer slurries. AB - We studied the metabolic fate of bromacil in anaerobic aquifer slurries held under denitrifying, sulfate-reducing, or methanogenic conditions. Liquid chromatograhy-mass spectrometry of the slurries confirmed that bromacil was debrominated under methanogenic conditions but was not degraded under the other incubation conditions. This finding extends the range of aryl reductive dehalogenation reactions to include nitrogen heterocyclic compounds. PMID- 16348104 TI - Carbon Flow in Mercury Biomethylation by Desulfovibrio desulfuricans. AB - Radiocarbon incorporation from pyruvate and serine into monomethylmercury by Desulfovibrio desulfuricans was consistent with the proposal that the methyl group originates from C-3 of serine. Immunodiagnostic assays measured 4 to 35 mug of tetrahydrofolate and 58 to 161 ng of cobalamin or a closely related cobalt porphyrin per g of cell protein in D. desulfuricans. The light-reversible inhibition of mercury methylation by propyl iodide in D. desulfuricans indicates methyl transfer by a cobalt porphyrin. PMID- 16348105 TI - Incidence of coliphage in potable water supplies. AB - [This corrects the article on p. 1632 in vol. 54.]. PMID- 16348106 TI - Characterization of a Symbiotic Coculture of Clostridium thermohydrosulfuricum YM3 and Clostridium thermocellum YM4. AB - Clostridium thermohydrosulfuricum YM3 and C. thermocellum YM4 were isolated from a coculture which was obtained from an enrichment culture inoculated with volcanic soil in Izu Peninsula, Japan. Strain YM3 had advantages over reported C. thermohydrosulfuricum strains in that it fermented inulin and could accumulate ethanol up to 1.3% (wt/vol). The highest ethanol yield obtained was 1.96 mol/mol of anhydroglucose unit in cellobiose. Strain YM4 had features different from those reported in C. thermocellum strains: it formed spores rarely (at a frequency of <10), it required CO(2) and Na(2)CO(3) for growth, and it fermented sucrose. Strain YM4 completely decomposed 1% Avicel within 25 h when the inoculum constituted 2% of the culture medium volume, and it produced 0.22 U of Avicelase and 2.21 U of carboxymethylcellulase per ml of the medium. The doubling times on Avicel, cellobiose, and glucose were 2.7, 1.1, and 1.6 h, respectively. Reconstructed cocultures of strains YM3 and YM4 were very stable and degraded Avicel more rapidly than did strain YM4 monoculture. Without yeast extract, neither microorganism was able to grow. However, the coculture grew on cellulose without yeast extract and produced ethanol in high yield. Moreover, cell-free spent culture broth of strain YM3 could replace yeast extract in supporting the growth of strain YM4. The symbiotic relationship of the two bacteria in cellulose fermentation is probably a case of mutualism. PMID- 16348107 TI - Characterization of palo podrido, a natural process of delignification in wood. AB - Chemical and morphological changes of incipient to advanced stages of palo podrido, an extensively delignified wood, and other types of white rot decay found in the temperate forests of southern Chile were investigated. Palo podrido is a general term for white rot decay that is either selective or nonselective for the removal of lignin, whereas palo blanco describes the white decayed wood that has advanced stages of delignification. Selective delignification occurs mainly in trunks of Eucryphia cordifolia and Nothofagus dombeyi, which have the lowest lignin content and whose lignins have the largest amount of beta-aryl ether bonds and the highest syringyl/guaiacyl ratio of all the native woods included in this study. A Ganoderma species was the main white rot fungus associated with the decay. The structural changes in lignin during the white rot degradation were examined by thioacidolysis, which revealed that the beta-aryl ether-linked syringyl units were more specifically degraded than the guaiacyl ones, particularly in the case of selective delignification. Ultrastructural studies showed that the delignification process was diffuse throughout the cell wall. Lignin was first removed from the secondary wall nearest the lumen and then throughout the secondary wall toward the middle lamella. The middle lamella and cell corners were the last areas to be degraded. Black manganese deposits were found in some, but not all, selectively delignified samples. In advanced stages of delignification, almost pure cellulose could be found, although with a reduced degree of polymerization. Cellulolytic enzymes appeared to be responsible for depolymerization. A high brightness and an easy refining capacity were found in an unbleached pulp made from selectively delignified N. dombeyi wood. Its low viscosity, however, resulted in poor resistance properties of the pulp. The last stage of degradation (i.e., decomposition of cellulose-rich secondary wall layers) resulted in a gelatinlike substance. Ultrastructural and chemical analyses of this substance showed the matrix to have no microfibrillar structure characteristic of woody cell walls but to still be rich in glucan. PMID- 16348108 TI - Effects of toxic substances on natural bacterial assemblages determined by means of [h]thymidine incorporation. AB - The effects of 3,5-dichlorophenol, 2,4-dinitrophenol, and potassium dichromate on natural bacterial assemblages were examined by means of [H]thymidine incorporation into trichloroacetic acid-insoluble material. Results from a large number of coastal marine and freshwater samples suggest the following. (i) The effects of the three toxicants included reductions in the bacterial cell number as well as changes in rates of [H]thymidine incorporation and in [H]thymidine incorporation per cell. The concentrations that inhibited [H]thymidine incorporation by 50% ranged from 3 to 11 mg liter for 3,5-dichlorophenol, 6 to 10 mg liter for 2,4-dinitrophenol, and 21 to 123 mg liter for potassium dichromate, with a tendency to higher values in bacterial assemblages from more eutrophic environments. (ii) The effects of 3,5-dichlorophenol and potassium dichromate determined by [H]leucine incorporation into bacterial protein were similar or larger than those obtained from [H]thymidine incorporation. (iii) Two to four hours of exposure to the toxicants was necessary before stable maximum effects were found in [H]thymidine incorporation. (iv) Storage of natural environmental samples should be avoided, since tests with water stored for 1 to 3 days sometimes produced results different from results obtained from in situ tests. (v) The effects of 3,5-dichlorophenol, 2,4-dinitrophenol, and potassium dichromate on natural bacterial assemblages were relatively constant during periods with different growth rates in the assemblages, during various periods of the year, and between samples from freshwater and marine localities. With some precautions, [H]thymidine incorporation can be used as a quick and sensitive method for determining the effects of toxicants on aquatic bacterial assemblages from natural environmental samples. PMID- 16348109 TI - Construction of a Symbiotically Effective Strain of Rhizobium leguminosarum bv. trifolii with Increased Nodulation Competitiveness. AB - Genes involved in nodulation competitiveness (tfx) were inserted by marker exchange into the genome of the effective strain Rhizobium leguminosarum bv. trifolii TA1. Isogenic strains of TA1 were constructed which differed only in their ability to produce trifolitoxin, an antirhizobial peptide. Trifolitoxin production by the ineffective strain R. leguminosarum bv. trifolii T24 limited nodulation of clover roots by trifolitoxin-sensitive strains of R. leguminosarum bv. trifolii. The trifolitoxin-producing exconjugant TA1::10-15 was very competitive for nodulation on clover roots when coinoculated with a trifolitoxin sensitive reference strain. The nonproducing exconjugant TA1::12-10 was not competitive for nodule occupancy when coinoculated with the reference strain. Tetracycline sensitivity and Southern analysis confirmed the loss of vector DNA in the exconjugants. Trifolitoxin production by TA1::10-15 was stable in the absence of selection pressure. Transfer of tfx to TA1 did not affect nodule number or nitrogenase activity. These experiments represent the first stable genetic transfer of genes involved in nodulation competitiveness to a symbiotically effective Rhizobium strain. PMID- 16348110 TI - Thiobacillus cuprinus sp. nov., a Novel Facultatively Organotrophic Metal Mobilizing Bacterium. AB - Five strains of mesophilic, facultatively organotrophic, ore-leaching eubacteria were isolated from solfatara fields in Iceland and a uranium mine in the Federal Republic of Germany. The new organisms are aerobic gram-negative rods. They can use sulfidic ores or elemental sulfur as sole energy source, indicating that they belong to the genus Thiobacillus. Alternatively, they grow on organic substrates such as yeast extract, peptone, and pyruvate. In contrast to the other leaching bacteria known so far, the new isolates are unable to oxidize ferrous iron. They consist of extreme and moderate acidophiles growing optimally at pH 3 and 4, respectively. The extreme acidophiles showed leaching characteristics similar to those shown by Thiobacillus ferrooxidans, while the moderate acidophiles exhibited a pronounced preference for copper leaching on some chalcopyrite ores. The G+C content of the DNA is between 66 and 69 mol%, depending on the isolate. In DNA-DNA hybridization experiments, the new strains showed homologies among each other of >70%, indicating that they belong to the same species. No significant DNA homology to Thiobacillus reference strains was detectable. Therefore, the new isolates represent a new species of Thiobacillus, which we name Thiobacillus cuprinus. Isolate Ho5 is designated as the type strain (DSM 5495). PMID- 16348111 TI - Mineralization of surfactants by the microbiota of submerged plant detritus. AB - In wetlands and canopied bodies of water, plant detritus is an important source of carbon and energy. Detrital materials possess a large surface area for sorption of dissolved organics and are colonized by a large and diverse microbiota. To examine the biodegradation of surfactants by these microorganisms, submerged oak leaves were obtained from a laundromat wastewater pond, its overflow, and a pristine control pond. Leaves were cut into disks and incubated in sterile water amended with 50 mug of C-labeled linear alkylbenzene sulfonate (LAS), linear alcohol ethoxylate, stearyltrimethyl ammonium chloride, distearyldimethyl ammonium chloride, benzoic acid, or mixed amino acids per liter. Sorption of the test compounds to the detritus and evolution of CO(2) were followed with time. All of the compounds sorbed to the detritus to various degrees, with LAS and stearyltrimethyl ammonium chloride the most sorptive and benzoic acid the least. All compounds were mineralized without a lag. With leaves from the laundromat wastewater pond, half-lives were 12.6 days for LAS, 8.4 days for linear alcohol ethoxylate, 14.2 days for stearyltrimethyl ammonium chloride, 1.0 days for benzoic acid, and 2.7 days for mixed amino acids. Mineralization of LAS and linear alcohol ethoxylate by control pond leaves was slower and exhibited an S-shaped rather than a typical first-order pattern. This study shows that detritus represents a significant site of surfactant removal in detritus-rich systems. PMID- 16348112 TI - Occurrence of a Thiothrix sp. Attached to Mayfly Larvae and Presence of Parasitic Bacteria in the Thiothrix sp. AB - Larvae of the mayfly (Drunella grandis [Eaton]) from Diamond Fork Creek, Utah, were covered with a heavy growth of the sulfide-oxidizing bacterium Thiothrix. The bacterium did not seem to harm the mayfly, but the Thiothrix trichomes were parasitized by three morphologically distinct bacteria, two of which were cytoplasmic and one of which was probably periplasmic. At least two of the parasites destroyed the cytoplasmic contents of the Thiothrix sp., thus killing the host cell. Attempts to obtain the parasites in pure culture were unsuccessful. PMID- 16348113 TI - Purification and Partial Characterization of a Prolyl-Dipeptidyl Aminopeptidase from Lactobacillus helveticus CNRZ 32. AB - X-prolyl-dipeptidyl aminopeptidase, which hydrolyzed Gly-Pro-p-nitroanilide (relative activity [RA] = 100%) and Arg-Pro-p-nitroanilide (RA, 130%), was purified to homogeneity from the cell extract of Lactobacillus helveticus CNRZ 32. The enzyme also hydrolyzed Ala-Pro-Gly (RA, 11%) and Ala-Ala-p-nitroanilide (RA, 2%) but was not active on Ala-Leu-Ala, dipeptides, and endopeptidase and carboxypeptidase substrates. The enzyme was purified 145-fold by streptomycin sulfate precipitation, ammonium sulfate fractionation, and a series of column chromatographies on DEAE-cellulose, arginine-Sepharose 4B, and glycyl-prolyl-AH Sepharose 4B. The purified enzyme appeared as a single band on native polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoreses and had a molecular weight of 72,000. Optima for activity by the purified enzyme were pH 7.0 and 40 degrees C. The enzyme was incubated at 40 degrees C for 15 min with various metal ions. It was activated by Mg (2.5 mM), Ca (0.1 to 2.5 mM), Na (10 to 50 mM), and K (10 to 50 mM) and was inhibited by Hg (0.1 to 2.5 mM), Cu (0.1 to 2.5 mM), and Zn (0.1 to 2.5 mM). Enzyme activity was partially inhibited by EDTA (1.0 mM, 20 h at 40 degrees C), 1,10-phenanthroline (1.0 mM, 15 min at 40 degrees C), phenylmethylsulfonyl fluoride (1.0 mM), N-ethylmaleimide (1.0 mM), and iodoacetate (1.0 mM). It was completely inhibited by diisopropyl fluorophosphate (1.0 mM, 2 h at 40 degrees C) and p-chloromercuribenzoate (1.0 mM, 15 min at 40 degrees C). The enzyme was not affected by dithioerythritol (1.0 to 10 mM). PMID- 16348114 TI - Effect of Environmental Conditions on Extracellular Protease Activity in Lignolytic Cultures of Phanerochaete chrysosporium. AB - Two different types of extracellular protease activity were identified in the culture fluid of Phanerochaete chrysosporium wild-type BKM-F grown in submerged batch culture on N-limited media. The first activity, which appears to be inherent to the active growth phase, displayed a maximum on day 2 and decreased to a very low level on day 4. The second activity, which appeared at day 8 following the peak of ligninase activity, seems to be characteristic of late secondary metabolism and is stimulated by carbon starvation. Cultures started with half the amount of glucose of other cultures showed a remarkably earlier development of secondary activity. In contrast, the fed-batch addition of glucose started when ligninase activity was at a maximum (day 6) completely repressed secondary protease activity and enhanced ligninase production. The addition of exogenous veratryl alcohol increased the level of secondary protease activity, whereas the oxygen supply pattern significantly affected both the time course and the level of overall proteolytic activity. The addition of phenylmethylsulfonyl fluoride to growing cultures (0, 1, or 6 days) diminished overall protease activity, while it significantly enhanced ligninase activity. In all cases, the time courses of protease and ligninase activities were negatively correlated, indicating that protease activity promotes the decline of ligninase activity in batch culture. PMID- 16348115 TI - Increased sporulation of vesicular-arbuscular mycorrhizal fungi by manipulation of nutrient regimens. AB - Adjustment of pot culture nutrient solutions increased root colonization and sporulation of vesicular-arbuscular mycorrhizal (VAM) fungi. Paspalum notatum Flugge and VAM fungi were grown in a sandy soil low in N and available P. Hoagland nutrient solution without P enhanced sporulation in soil and root colonization of Acaulospora longula, Scutellospora heterogama, Gigaspora margarita, and a wide range of other VAM fungi over levels produced by a tap water control or nutrient solutions containing P. However, Glomus intraradices produced significantly more spores in plant roots in the tap water control treatment. The effect of the nutrient solutions was not due solely to N nutrition, because the addition of NH(4)NO(3) decreased both colonization and sporulation by G. margarita relative to levels produced by Hoagland solution without P. PMID- 16348116 TI - Monoclonal Antibodies to Ferric Pseudobactin, the Siderophore of Plant Growth Promoting Pseudomonas putida B10. AB - Monoclonal antibodies to ferric pseudobactin, the siderophore (microbial iron transport agent) of plant growth-promoting Pseudomonas putida B10, have been developed. Three immunoglobulin G subclass 1-type monoclonal antibodies have been characterized. Each antibody appears to be unique on the basis of their reactions with ferric pseudobactin and with culture supernatants from other pseudomonads. None of the three cross-reacts with ferric pseudobactin-type siderophores produced by seven other pseudomonads. However, P. aeruginosa ATCC 15692 and P. fluorescens ATCC 17400 produced relatively high-molecular-mass compounds (mass greater than approximately 30,000 daltons) that did react with the antibodies. The compound from P. aeruginosa was not iron regulated, while the compound from P. fluorescens was produced only under iron-limiting conditions. A competitive assay using these antibodies has a detection limit of 5 x 10 mol of ferric pseudobactin. This is, to our knowledge, the first report of monoclonal antibodies reactive with siderophores. PMID- 16348117 TI - Production of bakers' yeast in cheese whey ultrafiltrate. AB - A process for the production of bakers' yeast in whey ultrafiltrate (WU) is described. Lactose in WU was converted to lactic acid and galactose by fermentation. Streptococcus thermophilus was selected for this purpose. Preculturing of S. thermophilus in skim milk considerably reduced its lag. Lactic fermentation in 2.3x-concentrated WU was delayed compared with that in unconcentrated whey, and fermentation could not be completed within 60 h. The growth rate of bakers' yeast in fermented WU differed among strains. The rate of galactose utilization was similar for all strains, but differences in lactic acid utilization occurred. Optimal pH ranges for galactose and lactic acid utilization were 5.5 to 6.0 and 5.0 to 5.5, respectively. The addition of 4 g of corn steep liquor per liter to fermented WU increased cell yields. Two sources of nitrogen were available for growth of Saccharomyces cerevisiae: amino acids (corn steep liquor) and ammonium (added during the lactic acid fermentation). Ammonium was mostly assimilated during growth on lactic acid. This process could permit the substitution of molasses by WU for the industrial production of bakers' yeast. PMID- 16348118 TI - Copper resistance gene homologs in pathogenic and saprophytic bacterial species from tomato. AB - Copper-resistant strains of Xanthomonas campestris pv. vesicatoria, Pseudomonas cichorii, Pseudomonas putida, Pseudomonas fluorescens, and a yellow Pseudomonas sp. were isolated from tomato plants or seeds. In Southern hybridizations, DNA from each strain showed homology with the copper resistance (cop) operon previously cloned from Pseudomonas syringae pv. tomato PT23. Homology was associated with plasmid and chromosomal DNA in X. compestris pv. vesicatoria, P. putida, and the yellow Pseudomonas sp. Homology was detected only in the chromosomal DNA of P. cichorii and P. fluorescens. Homology with cop was also detected in chromosomal DNA from copper-sensitive strains of P. cichorii, P. fluorescens, and P. syringae pv. tomato, suggesting that the cop homolog may be indigenous to certain Pseudomonas species and have some function other than copper resistance. No homology was detected in DNA from a copper-sensitive X. campestris pv. vesicatoria strain. Copper-inducible protein products were detected in each copper-resistant bacterium by immunoblot analysis with antibodies raised to the CopB protein from the cop operon. The role of the homologous DNA in copper resistance was confirmed for the X. campestris pv. vesicatoria strain by cloning and transferring the cop homolog to a copper sensitive strain of X. campestris pv. vesicatoria. The possibility and implications of copper resistance gene exchange between different species and genera of pathogenic and saprophytic bacteria on tomato plants are discussed. PMID- 16348119 TI - Stability of Markers Used for Identification of Two Rhizobium galegae Inoculant Strains after Five Years in the Field. AB - The stability of identification markers was examined for two Rhizobium galegae inoculant strains after 5 years in the field. The two strains are genetically closely related, but differ in their lipopolysaccharides. Strain HAMBI 540 has lipopolysaccharide of the rough type, whereas that of strain HAMBI 1461 is of the smooth type. The properties that were examined for 10 field isolates of each inoculant type were symbiotic phenotype, phage type, intrinsic antibiotic resistance, maximum growth temperature, lipopolysaccharide and total soluble protein patterns, immunological properties, DNA restriction profiles, and DNA hybridization patterns, which were determined by using nifHDK and recA sequences as probes. Of these properties, all remained stable in soil, with the exception of some variation in intrinsic antibiotic resistance and the acquisition of an extra EcoRI restriction fragment by one of the isolates. Thus, both the rough and the smooth lipopolysaccharide phenotypes persisted equally well in soil. PMID- 16348120 TI - Diel nitrogen fixation by cyanobacterial surface blooms in sanctuary lake, pennsylvania. AB - Diel nitrogen fixation studies were conducted with assemblages of cyanobacteria sampled from surface blooms on Sanctuary Lake, Pa. The studies were conducted between July and September of 1982 to 1985 by using the acetylene reduction technique. Assemblages with the lowest cell concentrations (0.9 x 10 to 1.0 x 10 cells per liter) exhibited nitrogen fixation activity throughout the day, with maximum fixation rates occurring in mid to late afternoon; fixation proceeded throughout the night at rates equivalent to 23 to 28% of the afternoon maximum. In studies conducted with the highest cell concentrations (3.7 x 10 to 6.7 x 10 cells per liter), fixation rates reached maximum values in mid to late morning. The rates declined rapidly throughout the midday period and subsequently ceased from late afternoon until sunrise on the following day. The afternoon decline and cessation of fixation exhibited by high cell concentrations correlated with photosynthetically induced low total CO(2) and supersaturating O(2) concentrations. The midday decline could be prevented and partially reversed by experimentally lowering O(2) and increasing total CO(2) concentrations. Under experimental conditions which simultaneously prevented supersaturating O(2) concentrations and maintained high total CO(2) availability, nitrogen fixation continued throughout the solar day, with maximum rates occurring at midday. These observations indicate that temporal changes in photosynthetic activity may affect diel fluctuations in nitrogen fixation. PMID- 16348121 TI - CO(2) Incorporation and 4-Hydroxy-2-Methylbenzoic Acid Formation during Anaerobic Metabolism of m-Cresol by a Methanogenic Consortium. AB - The metabolism of m-cresol by methanogenic cultures enriched from domestic sewage sludge was investigated. In the initial studies, bromoethanesulfonic acid was used to inhibit methane production. This led to the accumulation of 4.0 +/- 0.8 mol of acetate per mol of m-cresol metabolized. These results suggested that CO(2) incorporation occurred because each molecule of m-cresol contained seven carbon atoms, whereas four molecules of acetate product contained a total of eight carbon atoms. To verify this, [C]bicarbonate was added to bromoethanesulfonic acid-inhibited cultures, and those cultures yielded [C]acetate. Of the label recovered as acetate, 89% was found in the carboxyl position. Similar cultures fed [methyl-C]m-cresol yielded methyl-labeled acetate. A C-labeled transient intermediate was detected in cultures given either m-cresol and [C]bicarbonate or bicarbonate and [methyl-C]m-cresol. The intermediate was identified as 4-hydroxy-2-methylbenzoic acid. In addition, another metabolite was detected and identified as 2-methylbenzoic acid. This compound appeared to be produced only sporadically, and it accumulated in the medium, suggesting that the dehydroxylation of 4-hydroxy-2-methylbenzoic acid led to an apparent dead-end product. PMID- 16348122 TI - Enhanced yield of shiitake by saccharide amendment of the synthetic substrate. AB - Three experiments were performed to determine the effect of selected saccharides on mushroom yield and basidiome size of shiitake (Lentinula edodes) when grown on a synthetic substrate. Substrate formulations of sawdust, wheat bran, and millet were nonamended or amended with sucrose, fructose, or glucose. Addition of sucrose (0.6 to 1.2% [dry weight]) to the substrate stimulated mushroom yield by 11 to 20% or more. Addition of fructose at 1.2% and glucose at 0.6% resulted in similar yield increases. Most of the yield increase occurred on the first break. The substrate amended with 1.2% sucrose tended to have a more synchronous maturation for the second break resulting in fewer days when mushrooms were harvested. PMID- 16348123 TI - Bacterial production and growth rate estimation from [h]thymidine incorporation for attached and free-living bacteria in aquatic systems. AB - Production and specific growth rates of attached and free-living bacteria were estimated in an oligotrophic marine system, La Salvaje Beach, Vizcaya, Spain, and in a freshwater system having a higher nutrient concentration, Butron River, Vizcaya, Spain. Production was calculated from [methyl-H]thymidine incorporation by estimating specific conversion factors (cells or micrograms of C produced per mole of thymidine incorporated) for attached and free-living bacteria, respectively, in each system. Conversion factors were not statistically different between attached and free-living bacteria: 6.812 x 10 and 8.678 x 10 mug of C mol for free-living and attached bacteria in the freshwater system, and 1.276 x 10 and 1.354 x 10 mug of C mol for free-living and attached bacteria in the marine system. Therefore, use of a unique conversion factor for the mixed bacterial population is well founded. However, conversion factors were higher in the freshwater system than in the marine system. This could be due to the different trophic conditions of the two systems. Free-living bacteria contributed the most to production in the two systems (85% in the marine system and 67% in the freshwater system) because of their greater contribution to total biomass. Specific growth rates calculated from production data and biomass data were similar for attached and free-living bacteria. PMID- 16348124 TI - Osmoregulation in Rhizobium meliloti: Production of Glutamic Acid in Response to Osmotic Stress. AB - Rhizobium meliloti, like many other bacteria, accumulates high levels of glutamic acid when osmotically stressed. The effect was found to be proportional to the osmolarity of the growth medium. NaCl, KCI, sucrose, and polyethylene glycol elicited this response. The intracellular levels of glutamate and K began to increase immediately when cells were shifted to high-osmolarity medium. Antibiotics that inhibit protein synthesis did not affect this increase in glutamate production. Cells growing in conventional media at any stage in the growth cycle could be suspended in medium causing osmotic stress and excess glutamate accumulated. The excess glutamate did not appear to be excreted, and the intracellular level eventually returned to normal when osmotically stressed cells were suspended in low-osmolarity medium. A glt mutant lacking glutamate synthase and auxotrophic for glutamate accumulated excess glutamate in response to osmotic stress. Addition of isoleucine, glutamine, proline, or arginine stimulated glutamate accumulation to wild-type levels when the mutant cells were suspended in minimal medium with NaCl to cause osmotic stress. In both wild-type and mutant cells, inhibitors of transaminase activity, including azaserine and aminooxyacetate, reduced glutamate levels. The results suggest that the excess glutamate made in response to osmotic stress is derived from degradation of amino acids and transamination of 2-ketoglutarate. PMID- 16348125 TI - Role of Microniches in Protecting Introduced Rhizobium leguminosarum biovar trifolii against Competition and Predation in Soil. AB - The importance of microniches for the survival of introduced Rhizobium leguminosarum biovar trifolii cells was studied in sterilized and recolonized sterilized loamy sand and silt loam. The recolonized soils contained several species of soil microorganisms but were free of protozoa. Part of these soil samples was inoculated with the flagellate Bodo saltans, precultured on rhizobial cells. The introduced organisms were enumerated in different soil fractions by washing the soil, using a standardized washing procedure. With this method, free organisms and organisms associated with soil particles or aggregates >50 mum were separated. The total number of rhizobia was influenced slightly (silt loam) or not at all (loamy sand) by the recolonization with microorganisms or by the addition of flagellates alone. However, when both flagellates and microorganisms were present, numbers of rhizobia decreased drastically. This decrease was more than the sum of both effects separately. Nevertheless, populations of rhizobia were still higher than in natural soil. In the presence of flagellates, higher percentages of rhizobia and other microorganisms were associated with soil particles or aggregates >50 mum than in the absence of flagellates. In recolonized soils, however, the percentages of particle-associated rhizobia were lower than in soils not recolonized previous to inoculation. Thus, the presence of other microorganisms hindered rhizobial colonization of sites where they are normally associated with soil particles or aggregates. PMID- 16348126 TI - Characterization of naturally occurring atrazine-resistant isolates of the purple non-sulfur bacteria. AB - Six isolates of the purple non-sulfur bacteria, which upon primary isolation were naturally resistant to the herbicide atrazine, were characterized with respect to their taxonomic identity and the mechanism of their resistance. On the basis of electron microscopy, photopigment analysis, and other criteria, they were identified as strains of Rhodopseudomonas acidophila, Rhodopseudomonas palustris, or Rhodocyclus gelatinosus. These isolates exhibited degrees of atrazine resistance which ranged from 1.5 to about 4 times greater than that of cognate reference strains (American Type Culture Collection) tested. Furthermore, all of the reference strains tested were more intrinsically resistant to atrazine than was Rhodobacter sphaeroides. No unique plasmids which might encode for herbicide degradation or inactivation were found in these isolates. Resistance to the herbicide in these isolates was not the result of diminished binding of the herbicide to the L subunit of the bacterial reaction center. Differences in herbicide resistance among the various species of this group may be the result of compositional and chemical differences in the individual reaction centers. However, the increase in atrazine resistance for the isolates characterized in this study probably occurs by undefined mechanisms and not necessarily by changes in the binding of the herbicide to the L subunit of the photosynthetic reaction center. PMID- 16348127 TI - Chlorine Dioxide Gas Sterilization under Square-Wave Conditions. AB - Experiments were designed to study chlorine dioxide (CD) gas sterilization under square-wave conditions. By using controlled humidity, gas concentration, and temperature at atmospheric pressure, standard biological indicators (BIs) and spore disks of environmental isolates were exposed to CD gas. The sporicidal activity of CD gas was found to be concentration dependent. Prehumidification enhanced the CD activity. The D values (time required for 90% inactivation) of Bacillus subtilis subsp. niger ATCC 9372 BIs were estimated to be 1.5, 2.5, and 4.2 min when exposed to CD concentrations of 30, 15, and 7 mg/liter, respectively, at 23 degrees C and ambient (20 to 40%) relative humidity (RH). Survivor tailings were observed. Prehumidification of BIs to 70 to 75% RH in an environmental chamber for 30 min resulted in a D value of 1.6 min after exposure to a concentration of 6 to 7 mg of CD per liter at 23 degrees C and eliminated survivor tailing. Prolonging prehumidification at 70 to 75% RH for up to 16 h did not further improve the inactivation rate. Prehumidification by ultrasonic nebulization was found to be more effective than prehumidification in the environmental chamber, improving the D value to 0.55 min at a CD concentration of 6 to 7 mg/liter. Based on the current observations, CD gas is estimated, on a molar concentration basis, to be 1,075 times more potent than ethylene oxide as a sterilant at 30 degrees C. A comparative study showed B. subtilis var. niger BIs were more resistant than other types of BIs and most of the tested bacterial spores of environmental isolates. PMID- 16348128 TI - Purification and Characterization of an Aminopeptidase from Lactococcus lactis subsp. cremoris Wg2. AB - An aminopeptidase was purified to homogeneity from a crude cell extract of Lactococcus lactis subsp. cremoris Wg2 by a procedure that included diethyl aminoethane-Sephacel chromatography, phenyl-Sepharose chromatography, gel filtration, and high-performance liquid chromatography over an anion-exchange column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed a single protein band with a molecular weight of 95,000. The aminopeptidase was capable of degrading several peptides by hydrolysis of the N terminal amino acid. The peptidase had no endopeptidase or carboxypeptidase activity. The aminopeptidase activity was optimal at pH 7 and 40 degrees C. The enzyme was completely inactivated by the p-chloromecuribenzoate mersalyl, chelating agents, and the divalent cations Cu and Cd. The activity that was lost by treatment with the sulfhydryl-blocking reagents was restored with dithiothreitol or beta-mercapto-ethanol, while Zn or Co restored the activity of the 1,10-phenantroline-treated enzyme. Kinetic studies indicated that the enzyme has a relatively low affinity for lysyl-p-nitroanilide (K(m), 0.55 mM) but that it can hydrolyze this substrate at a high rate (V(max), 30 mumol/min per mg of protein). PMID- 16348129 TI - Additive Effects of Alcohols, Their Acidic By-Products, and Temperature on the Yeast Pachysolen tannophilus. AB - The effects of alcohols on the growth and fermentation of the yeast Pachysolen tannophilus were investigated at both 30 and 35 degrees C. Addition of alcohols to the culture medium decreased both the growth rate and the final cell yield in a dose-dependent manner, and this decrease was more severe at 35 degrees C. The concentration for 50% growth rate inhibition decreased as the chain length of the alcohol increased. In fermentations using a high initial cell density, production of acids was always observed when the medium was supplemented with alcohols. Supplementation of the culture medium with a short-chain alcohol plus the corresponding acid was shown to exert an additive deleterious effect on fermentation, and this effect increased with temperature. Production of acids was associated with the presence of alcohol dehydrogenase activity in cell extracts. PMID- 16348130 TI - Kinetics of sulfur oxidation at suboptimal temperatures. AB - Chemolithoautotrophic bacteria were enriched from mine water at incubation temperatures ranging from 4 to 46 degrees C, using elemental sulfur as a substrate in acid mineral salts media. Thiobacillus-type bacteria were successfully enriched for at all test temperatures except 46 degrees C. Changes in pH (-dpH/dt) were used to estimate the rate constants for the enrichment cultures. The rate constants yielded a linear Arrhenius plot, an activation energy of 65 kJ/mol, and a temperature coefficient (Q(10)) of 2.1 for the 4 to 37 degrees C temperature interval. PMID- 16348131 TI - Pathovar-Specific Antigens of Xanthomonas campestris pv. begoniae and X. campestris pv. pelargonii Detected with Monoclonal Antibodies. AB - Two monoclonal antibodies specific for lipopolysaccharide antigens of Xanthomonas campestris pv. begoniae and pv. pelargonii reacted with all of their respective pathovar strains and not with 130 strains of other xanthomonads or 89 nonxanthomonads tested. These results, as well as previous results, indicate that pathovar-specific monoclonal antibodies were readily generated to strains of X. campestris pathovars that generally infect single hosts. PMID- 16348132 TI - Purine Auxotrophic Mutants with Altered Spore Color in Streptomyces azureus. ATCC 14921. AB - A purine auxotroph with pale purple spores and a derivative with yellowish orange spores were obtained from the wild-type strain of Streptomyces azureus ATCC 14921, which has bluish green spores. The changed color or pigmentation in the mutants was limited to the spores. They accumulated AIR (5'-phosphoribosyl-5 aminoimidazole) due to the lack of AIR carboxylase activity. PMID- 16348134 TI - Ecology of Vibrio mimicus in Aquatic Environments. AB - [This corrects the article on p. 2077 in vol. 55.]. PMID- 16348133 TI - Direct Selection of Clostridium acetobutylicum Fermentation Mutants by a Proton Suicide Method. AB - Clostridium acetobutylicum ATCC 10132 mutants altered in acetic acid synthesis or in the shift to solventogenesis were directly selected by a proton suicide method after mutagenic treatment, by using bromide and bromate as selective agents. The mutants were characterized according to their solvent and acid production. On the selection plates they differed in colony phenotype from the parent strain. PMID- 16348135 TI - Improved method of enumeration of attached bacteria for study of fluctuation in the abundance of attached and free-living bacteria in response to diel variation in seawater turbidity. AB - Sample preparation for enumerating attached bacteria in turbid seawater by epifluorescence microscopy was improved by treating samples with a surfactant (Tween 80) followed by sonication. With optimal treatment with Tween 80 (final concentration, 10 ppm [10 mug/ml]) and sonication, as many as 10 times more attached bacteria were enumerated from turbid seawater relative to the number enumerated from an untreated control. Dispersion of bacteria by sonication alone resulted in the enumeration of only 42 to 72% of the attached bacteria. By this technique, fluctuations in the number of attached and free-living bacteria were determined in water from Aransas Pass, Tex., where surface sediments are resuspended on a regular basis by tidal currents. The abundance of attached bacteria increased in proportion to the seawater turbidity that resulted from sediment resuspension. The variation in abundance of free-living bacteria was not directly related to seawater turbidity. However, the magnitude of fluctuation in the abundance of free-living bacteria was related to the extent of turbidity variation during diurnal tides. PMID- 16348136 TI - Nitrogen fixation dynamics of two diazotrophic communities in mono lake, california. AB - Two types of diazotrophic microbial communities were found in the littoral zone of alkaline hypersaline Mono Lake, California. One consisted of anaerobic bacteria inhabiting the flocculent surface layers of sediments. Nitrogen fixation (acetylene reduction) by flocculent surface layers occurred under anaerobic conditions, was not stimulated by light or by additions of organic substrates, and was inhibited by O(2), nitrate, and ammonia. The second community consisted of a ball-shaped association of a filamentous chlorophyte (Ctenocladus circinnatus) with diazotrophic, nonheterocystous cyanobacteria, as well as anaerobic bacteria (Ctenocladus balls). Nitrogen fixation by Ctenocladus balls was usually, but not always, stimulated by light. Rates of anaerobic dark fixation equaled those in the light under air. Fixation in the light was stimulated by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea and by propanil [N-(3,4 dichlorophenyl)propanamide]. 3-(3,4-Dichlorophenyl)-1,1-dimethyl urea-elicited nitrogenase activity was inhibited by ammonia (96%) and nitrate (65%). Fixation was greatest when Ctenocladus balls were incubated anaerobically in the light with sulfide. Dark anaerobic fixation was not stimulated by organic substrates in short-term (4-h) incubations, but was in long-term (67-h) ones. Areal estimates of benthic N(2) fixation were measured seasonally, using chambers. Highest rates ( approximately 29.3 mumol of C(2)H(4) m h) occurred under normal diel regimens of light and dark. These estimates indicate that benthic N(2) fixation has the potential to be a significant nitrogen source in Mono Lake. PMID- 16348137 TI - Bioconversion of cinnamic Acid to acetophenone by a pseudomonad: microbial production of a natural flavor compound. AB - A mutant derivative of a novel pseudomonad isolated from the soil accumulated acetophenone when supplied with cinnamic acid. The microorganism has been identified as an unclassified Pseudomonas sp., similar to Pseudomonas acidovorans. Mass spectrum analysis of the product acetophenone derived from catabolism of cinnamic acid in the presence of O(2) or H(2)O supported the conclusion that cinnamic acid degradation is initiated by addition of water to the double bond of the side chain, followed by dehydrogenation to generate 3-keto 3-phenylpropionic acid. The intermediate 3-keto-3-phenylpropionic acid is accumulated in cultures of the mutant during active cinnamic acid catabolism. However, this intermediate is unstable so a portion of it spontaneously decarboxylates to form acetophenone. Neither 3-keto-3-phenylpropionic acid nor acetophenone is a precedented intermediate in cinnamic acid degradation. Isolation of the novel strain and mutant provide the rudiments for a process to produce natural acetophenone by biotransformation of natural cinnamic acid. PMID- 16348138 TI - Effects of jet fuel spills on the microbial community of soil. AB - Hydrocarbon residues, microbial numbers, and microbial activity were measured and correlated in loam soil contaminated by jet fuel spills resulting in 50 and 135 mg of hydrocarbon g of soil. Contaminated soil was incubated at 27 degrees C either as well-aerated surface soil or as poorly aerated subsurface soil. In the former case, the effects of bioremediation treatment on residues, microbial numbers, and microbial activity were also assessed. Hydrocarbon residues were measured by quantitative gas chromatography. Enumerations included direct counts of metabolically active bacteria, measurement of mycelial length, plate counts of aerobic heterotrophs, and most probable numbers of hydrocarbon degraders. Activity was assessed by fluorescein diacetate (FDA) hydrolysis. Jet fuel disappeared much more rapidly from surface soil than it did from subsurface soil. In surface soil, microbial numbers and mycelial length were increased by 2 to 2.5 orders of magnitude as a result of jet fuel contamination alone and by 3 to 4 orders of magnitude as a result of the combination of jet fuel contamination and bioremediation. FDA hydrolysis was stimulated by jet fuel and bioremediation, but was inhibited by jet fuel alone. The latter was traced to an inhibition of the FDA assay by jet fuel biodegradation products. In subsurface soil, oxygen limitation strongly attenuated microbial responses to jet fuel. An increase in the most probable numbers of hydrocarbon degraders was accompanied by a decline in other aerobic heterotrophs, so that total plate counts changed little. The correlations between hydrocarbon residues, microbial numbers, and microbial activity help in elucidating microbial contributions to jet fuel elimination from soil. PMID- 16348139 TI - Bioremediation potential of terrestrial fuel spills. AB - A bioremediation treatment that consisted of liming, fertilization, and tilling was evaluated on the laboratory scale for its effectiveness in cleaning up a sand, a loam, and a clay loam contaminated at 50 to 135 mg g of soil by gasoline, jet fuel, heating oil, diesel oil, or bunker C. Experimental variables included incubation temperatures of 17, 27, and 37 degrees C; no treatment; bioremediation treatment; and poisoned evaporation controls. Hydrocarbon residues were determined by quantitative gas chromatography or, in the case of bunker C, by residual weight determination. Four-point depletion curves were obtained for the described experimental variables. In all cases, the disappearance of hydrocarbons was maximal at 27 degrees C and in response to bioremediation treatment. Poisoned evaporation controls underestimated the true biodegradation contribution, but nevertheless, they showed that biodegradation makes only a modest contribution to gasoline disappearance from soil. Bunker C was found to be structurally recalcitrant, with close to 80% persisting after 1 year of incubation. The three medium distillates, jet fuel, heating oil, and diesel oil, increased in persistence in the listed order but responded well to bioremediation treatment under all test conditions. With bioremediation treatment, it should be possible to reduce hydrocarbons to insignificant levels in contaminated soils within one growing season. PMID- 16348140 TI - Identification of Phytophthora citrophthora with Cloned DNA Probes. AB - Two different DNA fragments, one of 2.9 kilobases and the other of 5.1 kilobases, were cloned from Phytophthora citrophthora and showed no homology with DNA from plants and other related fungi. These DNA probes hybridized with DNA from 12 different P. citrophthora isolates obtained from a variety of hosts but did not hybridize with DNA from 6 P. citrophthora isolates obtained from cacao. Southern blot analysis revealed that the probes contained repetitive DNA, and restriction fragment length polymorphisms were identified among several P. citrophthora isolates. Of the isolates tested, two major groups were observed whose genetic similarity correlated with geographical distribution. One of the DNA probes was used to detect P. citrophthora growing from infected citrus roots incubated on semiselective medium. P. citrophthora was not detected by a hybridization assay of total DNA extracted directly from infected roots. PMID- 16348141 TI - Effects of Metals on Streptomyces coelicolor Growth and Actinorhodin Production. AB - Actinorhodin production by Streptomyces coelicolor was used as a model system to study the effects of metals on growth and polyketide synthesis in a streptomycete. Numerous metals were tested in cultures grown in liquid media. Mercury and cadmium were highly toxic, and copper, nickel, and lead were less so, but all tended to inhibit both growth and antibiotic synthesis to a similar extent. Unexpectedly, manganese, cobalt, zinc, and, to a lesser extent, chromium caused complex effects that in general resulted in some enhancement of growth yield but a reduction in antibiotic titers. These complex effects meant that cobalt, manganese, and zinc had lower 50% inhibitory concentrations for antibiotic yields compared with those for biomass. The physiologically active divalent cations calcium and magnesium were also tested. Calcium at high concentrations was particularly effective in reducing antibiotic titers and enhancing growth yields. By adding calcium at different phases of growth, it could be demonstrated that it was most effective in reducing the antibiotic yield when added during the early growth phase. Addition during the antibiotic producing phase resulted in little reduction of final actinorhodin titers. PMID- 16348142 TI - Physiological and Morphological Changes Induced by Nutrient Limitation of Pseudomonas fluorescens 378 in Continuous Culture. AB - Pseudomonas fluorescens 378 was studied in continuous culture at a dilution rate of 0.05 or 0.15 h and under a limitation of carbon/energy, nitrogen, phosphorus, iron(III), or oxygen. Cultures were examined for nutritional consumption, production of biosurfactant AP-6 and lipase, and electron microscopy morphology. Morphological features were lysis and plasmolysis of the cells, vacuoles in the cells, granules in cell nuclei, and DNA coagulation during transmission electron microscopy preparation. Biosurfactant and lipase production were lost after 8 to 15 retention times, but under iron limitation and at low dilution rate they were maintained for more than 30 retention times. Consumption of nutrients varied between different cultures. Between 2.4 and 6.0 g of succinic acid per g (dry weight) was consumed; the highest value was obtained under phosphorus limitation. The uptake of nitrogen was mostly about 0.16 g/g (dry weight), and that of phosphorus varied between 13 and 58 mg/g (dry weight). Phosphorus-limited cells reduced their phosphorus consumption by at least 50% compared with other limitations. Cell morphology varied among different cultures. Up to 25% cell lysis occurred at the higher dilution rate. The frequencies of plasmolysis varied between 0 and 85%. Granules in nuclei were found in 65 to 100% of the cells. Vacuoles appeared mostly in low numbers, but at the lower dilution rate under phosphorus or iron limitation the frequencies increased to between 25 and 85%. At high dilution rate, the DNA coagulated in 30 to 70% of the cells. Multivariate data analysis demonstrated a general difference between the two tested dilution rates; i.e., both nutritional and morphological features differed more between the two tested dilution rates than between the different limitations. Cultures at the lower dilution rate changed more with time; this was especially pronounced for phosphorus or iron limitation. The data analysis also showed a correlation between plasmolysis or vacuoles in the cells and an increased carbon uptake under phosphorus limitation. PMID- 16348143 TI - Reduction of Cupric Ions with Elemental Sulfur by Thiobacillus ferrooxidans. AB - In anaerobic or aerobic conditions in the presence of 5 mM sodium cyanide, an inhibitor of iron oxidase, cupric ion (Cu) was reduced enzymatically with elemental sulfur (S) by washed intact cells of Thiobacillus ferrooxidans AP19-3 to give cuprous ion (Cu). The rate of Cu reduction was proportional to the concentrations of S and Cu added to the reaction mixture. The pH optimum for the cupric ion-reducing system was 5.0, and the activity was completely destroyed by 10-min incubation of cells at 70 degrees C. The activity of Cu reduction with S by this strain was strongly inhibited by inhibitors of hydrogen sulfide: ferric ion oxidoreductase (SFORase), such as alpha,alpha'-dipyridyl, 4,5-dihydroxy-m benzene disulfonic acid disodium salts, and diazine dicarboxylic acid bis-(N, N dimethylamide). A SFORase purified from this strain, which catalyzes oxidation of both hydrogen sulfide and S with Fe or Mo as an electron acceptor in the presence of glutathione, catalyzed a reduction of Cu by S, and the Michaelis constant of SFORase for Cu was 7.2 mM, indicating that a SFORase catalyzes the reduction of not only Fe and Mo but also Cu. PMID- 16348144 TI - Acetic Acid Bacterial Biota of the Pink Sugar Cane Mealybug, Saccharococcus sacchari, and Its Environs. AB - Saccharococcus sacchari is the primary colonizer of the developing "sterile" tissue between the leaf sheath and stem of sugar cane. The honeydew secreted by the mealybugs is acidic (about pH 3) and supports an atypical epiphytic microbiota dominated by acetobacter-like bacteria and acidophilic yeast species. However, Erwinia and Leuconostoc species predominate within the leaf sheath pocket region when the mealybugs die out. The unidentified acetobacters were readily isolated from S. sacchari throughout its life cycle and from other genera of mealybugs on sugar cane and various other plants, both above and below ground. No other insect present on sugar cane was a significant vector of acetic acid bacteria. The major factors restricting microbial diversity within the environs of mealybugs were considered to be yeast activity along with bacterial production of acetic acid, ketogluconic acids, and gamma-pyrones, in association with their lowering of pH. The microbial products may aid in suppressing the attack by the parasitic mold Aspergillus parasiticus on mealybugs but could act as attractants for the predatory fruit fly Cacoxenus perspicax. PMID- 16348145 TI - Biological Control of Damping-Off of Alfalfa Seedlings with Bacillus cereus UW85. AB - We explored the potential of biological control of alfalfa (Medicago sativa L.) seedling damping-off caused by Phytophthora megasperma f. sp. medicaginis by screening root-associated bacteria for disease suppression activity in a laboratory bioassay. A total of 700 bacterial strains were isolated from the roots of field-grown alfalfa plants by using Trypticase soy agar. A simple, rapid assay was developed to screen the bacteria for the ability to reduce the mortality of Iroquois alfalfa seedlings that were inoculated with P. megasperma f. sp. medicaginis zoospores. Two-day-old seedlings were planted in culture tubes containing moist vermiculite, and each tube was inoculated with a different bacterial culture. Sufficient P. megasperma f. sp. medicaginis zoospores were added to each tube to result in 100% mortality of control seedlings. Of the 700 bacterial isolates tested, only 1, which was identified as Bacillus cereus and designated UW85, reduced seedling mortality to 0% in the initial screen and in two secondary screens. Both fully sporulated cultures containing predominantly released spores and sterile filtrates of these cultures of UW85 were effective in protecting seedlings from damping-off; filtrates of cultures containing predominantly vegetative cells or endospores inside the parent cell had low biocontrol activity. Cultures grown in two semidefined media had significantly greater biocontrol activities than cultures grown in the complex tryptic soy medium. In a small-scale trial in a field infested with P. megasperma f. sp. medicaginis, coating seeds with UW85 significantly increased the emergence of alfalfa. The results suggest that UW85 may have potential as a biocontrol agent for alfalfa damping-off, thus providing an alternative to current disease control strategies. PMID- 16348146 TI - Inhibition of the fermentation of propionate to methane by hydrogen, acetate, and propionate. AB - Inhibition of the fermentation of propionate to methane and carbon dioxide by hydrogen, acetate, and propionate was analyzed with a mesophilic propionate acclimatized sludge that consisted of numerous flocs (size, 150 to 300 mum). The acclimatized sludge could convert propionate to methane and carbon dioxide stoichiometrically without accumulating hydrogen and acetate in a propionate minimal medium. Inhibition of propionate utilization by propionate could be analyzed by a second-order substrate inhibition model (shown below) given that the substrate saturation constant, K(s), was 15.9 muM; the substrate inhibition constant, K(i), was 0.79 mM; and the maximum specific rate of propionate utilization, q(m), was 2.15 mmol/g of mixed-liquor volatile suspended solids (MLVSS) per day: q(s) = q(m)S/[K(s) + S + (S/K(i))], where q(s) is the specific rate of propionate utilization and S is the initial concentration of undissociated propionic acid. For inhibition by hydrogen and acetate to propionate utilization, a noncompetitive product inhibition model was used: q(s) = q(m)/[1 + (P/K(p))], where P is the initial concentration of hydrogen or undissociated acetic acid and K(p) is the inhibition constant. Kinetic analysis gave, for hydrogen inhibition, K(p(H(2))) = 0.11 atm (= 11.1 kPa, 71.5 muM), q(m) = 2.40 mmol/g of MLVSS per day, and n = 1.51 and, for acetate inhibition, K(p(HAc)) = 48.6 muM, q(m) = 1.85 mmol/g of MLVSS per day, and n = 0.96. It could be concluded that the increase in undissociated propionic acid concentration was a key factor in inhibition of propionate utilization and that hydrogen and acetate cooperatively inhibited propionate degradation, suggesting that hydrogenotrophic and acetoclastic methanogens might play an important role in enhancing propionate degradation to methane and carbon dioxide. PMID- 16348147 TI - HCO(3) Fixation by Naturally Occurring Tufts and Pure Cultures of Thiothrix nivea. AB - Naturally occurring tufts of the mixotroph Thiothrix nivea blanketed the East Everglades (Dade County, Fla.) Chekika artesian well and runoff areas. The rate of HCO(3) fixation by these Thiothrix tufts was determined to be 14.0 +/- 5.4 nmol of HCO(3) per min per mg of dry weight, which reflected a growth rate of 5.0%/h. The addition of 10 mM glucose, ribose, acetate, or pyruvate or 0.05% Casamino Acids (Difco Laboratories, Detroit, Mich.) did not appear to alter the HCO(3) fixation rate. Whereas 1 mM acetate or 10 mM lactate, ethanol, glycerol, alpha-ketoglutarate, succinate, fumarate, or citrate slightly stimulated HCO(3) fixation, 5 to 10 mM malate inhibited HCO(3) fixation by 90%. Pure Thiothrix cultures isolated from Chekika fixed HCO(3) at rates as high as 29.9 +/- 2.8 nmol of HCO(3) per min per mg of dry weight in the presence of growth medium. Malate did not have a suppressive effect but rather slightly stimulated in vivo HCO(3) fixation. PMID- 16348148 TI - Bacterioplankton and organic carbon dynamics in the lower mesohaline chesapeake bay. AB - The mesohaline portion of the Chesapeake Bay is subject to annual summertime hypoxia and anoxia in waters beneath the pycnocline. This dissolved oxygen deficit is directly related to salinity-based stratification of the water column in combination with high levels of autochthonously produced organic matter and a very high abundance of metabolically active bacteria. Throughout the water column in the lower, mesohaline part of the bay, between the Potomac and Rappahannock rivers, near the southern limit of the mainstem anoxia, bacterial abundance often exceeded 10 x 10 cells per ml and bacterial production exceeded 7 x 10 cells per liter per day during summer. Bacterial biomass averaged 34% (range, 16 to 126%) of the phytoplankton biomass in summer. These values are equal to or greater than those found farther north in the bay, where the oxygen deficit is more severe. Seasonal variations in bacterial abundance and production were correlated with phytoplankton biomass (lag time, 7 to 14 days), particulate organic carbon and nitrogen, and particulate biochemical oxygen demand in spring; but during summer, they were significantly correlated only with dissolved biochemical oxygen demand. During summer, dissolved biochemical oxygen demand can account for 50 to 60% of the total biochemical oxygen demand throughout the water column and 80% in the bottom waters. There is a clear spring-summer seasonal shift in the production of organic matter and in the coupling of bacteria and autochthonous organic matter. The measurement of dissolved, microbially labile organic matter concentrations is crucial in understanding the trophic dynamics of the lower mesohaline part of the bay. The absolute levels of organic matter in the water column and the bacterial organic carbon relationships suggest that a lower bay source of organic matter fuels the upper mesohaline bay oxygen deficits. PMID- 16348149 TI - Die-away kinetic analysis of the capacity of epilithic and planktonic bacteria from clean and polluted river water to biodegrade sodium dodecyl sulfate. AB - The capacities of epilithic and planktonic river bacterial populations to degrade sodium dodecyl sulfate (SDS) in samples taken at two times during 1987 from one clean and four polluted sites in a South Wales river were estimated in die-away tests under simulated environmental conditions. There was a relatively slow disappearance of SDS in die-away tests for both planktonic and epilithic populations taken from the clean source site, as compared with those taken from the downstream polluted sites, for which the rate of biodegradation was accelerated, sometimes after an apparent initial lag period. The kinetic components contributing to the die-away curves were quantified by nonlinear regression analysis in which the experimental data were fitted to a variety of possible kinetic models. All samples except for one from the polluted sites best fitted a model which describes the biodegradation of SDS at concentrations well below its K(m) by bacteria whose growth is exponential and unaffected by the addition of a test substrate. Each sample from the clean source site fitted a different model, but there was generally little or no growth on endogenous carbon. A consideration of the numerical values of constants derived from the modeling of epilithic and planktonic populations from polluted sites indicated clearly that the biodegradative capacity of epilithic bacterial populations towards SDS is more stable than that of planktonic bacterial populations. PMID- 16348150 TI - Enhanced Growth of Wheat and Soybean Plants Inoculated with Azospirillum brasilense Is Not Necessarily Due to General Enhancement of Mineral Uptake. AB - The capacity of Azospirillum brasilense to enhance the accumulation of K, P, Ca, Mg, S, Na, Mn, Fe, B, Cu, and Zn in inoculated wheat and soybean plants was evaluated by using two different analytical methods with five A. brasilense strains originating from four distinct geographical regions. A Pseudomonas isolate from the rhizosphere of Zea mays seedlings was included as a control. All A. brasilense strains significantly improved wheat and soybean growth by increasing root and shoot dry weight and root surface area. The degree of plant response to inoculation varied among the different strains of A. brasilense. All strains were capable of colonizing roots, but the best root colonizer, Pseudomonas sp., had no effect on plant growth. The numbers of organisms of Brazilian strains Sp-245 and Sp-246 colonizing roots were similar regardless of the host plant. Numbers of organisms for the other strains were directly dependent on the host plant. The main feature characterizing mineral accumulation in inoculated plants was that all inoculation treatments changed the mineral balance of the plants, but in an inconsistent manner. Enhancement of mineral uptake by plants also varied among strains to a great extent and was directly dependent on the strain-plant combination; i.e., a strain capable of increasing accumulation of a particular ion in one plant species or cultivar often lacked the ability to do so in another. Minerals in inoculated plants were not evenly distributed in different plant tissues, and the changes varied among groups of plants within each bacterial strain inoculation treatment. We suggest that, although A. brasilense strains are capable of changing the mineral balance and content of plants, it is unlikely that this ability is a general mechanism responsible for plant improvement by A. brasilense. PMID- 16348151 TI - Toxicity of Irradiated Media for Xenorhabdus spp. AB - Bacterial isolates of the genus Xenorhabdus were shown to be extremely sensitive to photoproducts produced in a number of common media irradiated by fluorescent light. Two forms of toxic oxygen, hydrogen peroxide and superoxide radical, were produced in the media upon exposure to fluorescent light. The addition of pyruvate or catalase to the irradiated media eliminated the toxicity. The poor plating efficiencies previously reported for Xenorhabdus spp. are likely due to the uncontrolled exposure of media to ambient lighting. PMID- 16348152 TI - Use of robotics to transfer cultures on agar-containing media to fresh media. AB - The automation of transfer of microorganisms to fresh growth medium is described. The system was used to transfer our collection of about 1,500 bacteria and fungi without detectable contamination in a total time of 125 h. PMID- 16348153 TI - A Translation Fusion Product of Two Different Insecticidal Crystal Protein Genes of Bacillus thuringiensis Exhibits an Enlarged Insecticidal Spectrum. AB - Two truncated Bacillus thuringiensis crystal protein genes, belonging to the classes cryIA(b) and cryIC and both coding for insecticidal N-terminal fragments of the corresponding crystal proteins, were translationally fused. Expression of the gene fusion in Escherichia coli showed a biologically active protein with a toxicity spectrum that overlapped those of both contributing crystal proteins. PMID- 16348155 TI - Isolation and characterization of a pseudomonas strain that restricts growth of various phytopathogenic fungi. AB - The characterization of a novel Pseudomonas strain exhibiting antagonism towards many important corn fungal pathogens is presented. This strain was isolated from the caryopses of the grass Tripsacum dactyloides and was identified as Pseudomonas cepacia. The antagonistic activity is due to the production of an antifungal compound. The chromatographic properties of this partially purified compound isolated from growth medium differ from those reported previously for other pseudomonads. The suppression of the growth of economically important phytopathogens by this strain and by the partially purified compound indicates a potential biocontrol agent. PMID- 16348154 TI - Growth, Respiration, and Polypeptide Patterns of Bradyrhizobium sp. (Arachis) Strain 3G4b20 from Succinate- or Oxygen-Limited Continuous Cultures. AB - Succinate- or oxygen-limited continuous cultures were used to study the influences of different concentrations of dissolved oxygen and ammonia on the growth, respiration, and polypeptide patterns of Bradyrhizobium sp. (Arachis) strain 3G4b20. During succinate-limited growth, molar growth yields on succinate (Y(succ)) ranged from 38.9 to 44.4 g (dry weight) of cells mol of succinate and were not greatly influenced by changes in dilution rates or changes in the oxygen concentrations that we tested. Succinate, malate, and fumarate induced the highest rates of oxygen uptake in all of the steady states in which the supply rates of (NH(4))(2)SO(4) ranged between 322 and 976 mumol h. However, the amino acids aspartate, asparagine, and glutamate could also be used as respiratory substrates, especially when the (NH(4))(2)SO(4) supply rate was decreased to 29 mumol h. Glutamine-dependent respiration was seen only when the (NH(4))(2)SO(4) supply rate was 29 mumol h and thus appears to be under tight ammonia control. Nitrogenase activity was detected only when the culture was switched from a succinate-limited steady state to an oxygen-limited steady state. Comparison of major silver-stained proteins from three steady states by two-dimensional gel electrophoresis revealed that nearly 60% were affected by oxygen and 24% were affected by ammonia. These data are consistent with reports that oxygen has a major regulatory role over developmental processes in Rhizobium sp. and Bradyrhizobium sp. PMID- 16348156 TI - Enumeration of anaerobic chytridiomycetes as thallus-forming units: novel method for quantification of fibrolytic fungal populations from the digestive tract ecosystem. AB - An endpoint dilution procedure, based on the technique of most probable numbers, was developed to enumerate anaerobic chytridiomycetes as thallus-forming units. The method does not distinguish between zoospores and thalli, but does permit enumeration of fungal populations with respect to their ability to digest plant cell walls. Fibrolytic populations in batch culture, ruminal contents, and feces were compared by relating viable counts to the dry matter content of enumerated samples (i.e., thallus-forming units per gram of dry matter). Batch cultures of Neocallimastix sp. strain R1 grown on wheat straw were used to assess the enumeration procedure and to demonstrate the potential of the technique for quantification of anaerobic fungi in vivo. Determination of total ruminal contents from steers enabled the quantification of the entire population of fiber degrading anaerobic fungi in the reticulorumen. The enumeration procedure revealed substantial populations of fibrolytic anaerobic fungi in fresh and air dried feces. Populations in fresh feces were equivalent to those in ruminal contents, but declined exponentially with time in dry feces. Minimum values were obtained from dry feces 90 days after drying, and anaerobic fungi were detectable for up to 210 days thereafter. PMID- 16348157 TI - Isolation of Rhizobium leguminosarum (biovar trifolii) Strains from Ethiopian Soils and Symbiotic Effectiveness on African Annual Clover Species. AB - Strains of Rhizobium leguminosarum (biovar trifolii) isolated from two Ethiopian soils or obtained from a commercial source were evaluated for symbiotic effectiveness on five African annual clover species. Numerous Rhizobium trifolii strains that exhibited varying levels of symbiotic effectiveness were isolated from both soils (a nitosol and a vertisol), and it was possible to identify strains that were highly effective for each clover species. The soil isolates were, as a group, superior to the strains from the commercial source. Several R. trifolii strains were found to be effective on more than one clover species, and there appeared to be at least two and possibly three distinct cross-inoculation effectiveness groups. PMID- 16348158 TI - Identification and Plant Interaction of a Phyllobacterium sp., a Predominant Rhizobacterium of Young Sugar Beet Plants. AB - The second most abundant bacterium on the root surface of young sugar beet plants was identified as a Phyllobacterium sp. (Rhizobiaceae) based on a comparison of the results of 39 conventional identification tests, 167 API tests, 30 antibiotic susceptibility tests, and sodium dodecyl sulfate-polyacrylamide gel electrophoretic fingerprints of total cellular proteins with type strains of Phyllobacterium myrsinacearum and Phyllobacterium rubiacearum. It was found on 198 of 1,100 investigated plants between the 2nd and 10th leaf stage on three different fields in Belgium and one field in Spain. Densities ranged from 2 x 10 to 2 x 10 CFU/g of root. Five isolates exerted a broad-spectrum in vitro antifungal activity. DNA-DNA hybridizations showed that Phyllobacterium sp. does not contain DNA sequences that are homologous with the attachment genes chvA, chvB, the transferred-DNA (T-DNA) hormone genes iaaH and ipt from Agrobacterium tumefaciens, iaaM from A. tumefaciens and Pseudomonas savastanoi, or the nitrogenase genes nifHDK from Klebsiella pneumoniae. Phyllobacterium sp. produces indolylacetic acid in in vitro cultures and induces auxinlike effects when cocultivated with callus tissue of tobacco. When Phyllobacterium sp. was transformed with a Ti plasmid derivative, it gained the capacity to induce tumors on Kalanchoe daigremontiana. The potential role of Phyllobacterium sp. in this newly recognized niche is discussed. PMID- 16348160 TI - Transformation of Dibenzo-p-Dioxin by Pseudomonas sp. Strain HH69. AB - Dibenzo-p-dioxin was oxidatively cleaved by the dibenzofuran-degrading bacterium Pseudomonas sp. strain HH69 to produce minor amounts of 1-hydroxydibenzo-p-dioxin and catechol, while a 2-phenoxy derivative of muconic acid was formed as the major product. Upon acidic methylation, the latter yielded the dimethylester of cis, trans-2-(2-hydroxyphenoxy)-muconic acid. PMID- 16348159 TI - Metabolism of Dibenzofuran by Pseudomonas sp. Strain HH69 and the Mixed Culture HH27. AB - A Pseudomonas sp. strain, HH69, and a mixed culture, designated HH27, were isolated by selective enrichment from soil samples. The pure strain and the mixed culture grew aerobically on dibenzofuran as the sole source of carbon and energy. Degradation proceeded via salicylic acid which was branched into the gentisic acid and the catechol pathway. Both salicylic acid and gentisic acid accumulated in the culture medium of strain HH69. The acids were slowly metabolized after growth ceased. The enzymes responsible for their metabolism showed relatively low activities. Besides the above-mentioned acids, 2-hydroxyacetophenone, benzopyran 4-one (chromone), several 2-substituted chroman-4-ones, and traces of the four isomeric monohydroxydiben-zofurans were identified in the culture medium. 2,2',3 Trihydroxybiphenyl was isolated from the medium of a dibenzofuran-converting mutant derived from parent strain HH69, which can no longer grow on dibenzofuran. This gives evidence for a novel type of dioxygenases responsible for the attack on the biarylether structure of the dibenzofuran molecule. A meta-fission mechanism for cleavage of the dihydroxylated aromatic nucleus of 2,2',3 trihydroxybiphenyl is suggested as the next enzymatic step in the degradative pathway. PMID- 16348161 TI - Nitrogen Fixation in Continuous Culture with NH(4)Cl-Containing Media. AB - N balance and N dilution were determined from growth of Azospirillum brasilense Sp7 and two unidentified gram-negative nitrogen-fixing microorganisms in continuous culture supplied with NH(4)Cl. At the 1.1 and 2.2 mM NH(4)Cl steady states (N-to-C ratios of 1:68 and 1:34, respectively), the organisms grew with NH(4)Cl and N(2) as N sources simultaneously under carbon limitation. No ammonium could be detected in the supernatant of these cultures. PMID- 16348162 TI - Identification of deep-sea-sediment bacteria which produce tetrodotoxin. AB - Forty-nine bacterial strains were isolated from deep-sea sediments. Among them, 22 strains were shown by the tissue culture assay method to produce sodium channel blockers. For some strains, high-performance liquid chromatography analysis confirmed that the blocker was tetrodotoxin. Tetrodotoxin-producing bacteria seem to be widespread in marine sediment. PMID- 16348163 TI - Direct determination of nitrification in marine waters by using the short-lived radioisotope of nitrogen, N. AB - Biological oxidation of radiolabeled NH(4) (half-life = 10 min) was observed within minutes in assays of an estuarine ammonium oxidizer and in natural populations of nitrifiers in coastal waters. Our estimates of turnover of the ammonium pool and rates of nitrification based on experiments using N are consistent with previous values in the literature based on longer-term N tracer experiments or on indirect methods and thus provide corroboration for the estimates by other researchers. PMID- 16348164 TI - Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Pseudomonas fluorescens Proteases in Ultrahigh-Temperature-Treated Milk. AB - An inhibition enzyme-linked immunosorbent assay was developed to detect low levels of the proteases extracted from four strains of Pseudomonas fluorescens. The assay detected between 0.24 and 7.8 ng of protease per ml of ultrahigh temperature-treated milk and could be completed within 6 h. It could be used as a framework for a test system for quantifying spoilage proteases in dairy products. PMID- 16348165 TI - Composition and thermal properties of membrane lipids in cryptoendolithic lichen microbiota from antarctica. AB - In Antarctica there exists a cryptoendolithic microbiota which survives extremely low temperatures. Fatty acid analysis of the membrane phospholipids of this microbial community showed a predominance of polyunsaturated fatty acids. Artificial membranes made from the purified community phospholipids remained fluid to below -20 degrees C and had unusual hydration properties. PMID- 16348166 TI - Purification and Characterization of Glycerol Dehydratase from Lactobacillus reuteri. AB - A coenzyme B(12)-dependent glycerol dehydratase from Lactobacillus reuteri has been purified and characterized. The dehydratase has a molecular weight of approximately 200,000, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielded a single major band with a molecular weight of 52,000. K(m) values for substrates and coenzyme B(12) were in the millimolar and the submicromolar range, respectively. PMID- 16348167 TI - Dechlorination of chloroform by methanosarcina strains. AB - Dehalogenation of carbon tetrachloride, chloroform, and bromoform in pure cultures of Methanosarcina sp. strain DCM and Methanosarcina mazei S6 was demonstrated. The initial dechlorination product of chloroform was methylene chloride (dichloromethane), which accumulated transiently to about 70% of the added chloroform; trace amounts of chloromethane were also detected. The amount of chloroform dechlorinated per mole of methane produced was approximately 10 times greater than the ratio observed previously for tetrachloroethene dechlorination by these strains. The production of CO(2) from [C]chloroform and the absence of CH(4) imply that processes in addition to reductive dechlorination operate. PMID- 16348168 TI - Selective Plating Medium for Quantitative Recovery of Food-Borne Listeria monocytogenes. AB - [This corrects the article on p. 167 in vol. 56.]. PMID- 16348169 TI - Purification and Characterization of an Extracellular Acid Proteinase from the Ectomycorrhizal Fungus Hebeloma crustuliniforme. AB - Hebeloma crustuliniforme produced an extracellular acid proteinase in a liquid medium containing bovine serum albumin as the sole nitrogen source. The proteinase was purified 26-fold with 20% activity recovery and was shown to have a molecular weight of 37,800 (as indicated by sodium dodecyl sulfate polyacrylamide gel electrophoresis) and an isoelectric point of 4.8 +/- 0.2. The enzyme was most active at 50 degrees C and pH 2.5 against bovine serum albumin and was stable in the absence of substrates at temperatures up to 45 degrees C and pHs between 2.0 and 5.0. Pepstatin A, diazoacetyl-dl-norleucine methylester, metallic ions Fe and Fe, and phenolic acids severely inhibited the enzyme activity, while antipain, leupeptin, N-alpha-p-tosyl-l-lysine chloromethyl ketone, and trypsin inhibitor inhibited the activity moderately. The proteinase hydrolyzed bovine serum albumin and cytochrome c rapidly compared with casein and azocasein but failed to hydrolyze any of the low-molecular-weight peptide derivatives tested. PMID- 16348170 TI - Purification and Characterization of Three Chitosanase Activities from Bacillus megaterium P1. AB - Bacillus megaterium P1, a bacterial strain capable of hydrolyzing chitosan, was isolated from soil samples. Chitosan-degrading activity was induced by chitosan but not by its constituent d-glucosamine. Extracellular secretion of chitosanase reached levels corresponding to 1 U/ml under optimal conditions. Three chitosan degrading proteins (chitosanases A, B, and C) were purified to homogeneity. Chitosanase A (43 kilodaltons) was highly specific for chitosan and represented the major chitosan-hydrolyzing species. Chitosanases B (39.5 kilodaltons) and C (22 kilodaltons) corresponded to minor activities and possessed comparable specific activities toward chitosan, chitin, and cellulose. Chitosanase A was active from pH 4.5 to 6.5 and was stable on the basis of activity up to 45 degrees C. The optimum temperature for enzymatic chitosan hydrolysis was 50 degrees C. Kinetic studies on chitosanase A suggest that the enzyme is substrate inhibited. The apparent K(m) and V(max) determined at 22 degrees C and pH 5.6 were 0.8 mg/ml and 280 U/mg, respectively. End products of chitosan hydrolysis by each of the three chitosanases were identified as glucosamine oligomers, similar to those obtained for previously reported chitosanase digestions. PMID- 16348171 TI - Conversion of Wine Strains of Saccharomyces cerevisiae to Heterothallism. AB - A general method to convert homothallic strains of the yeast Saccharomyces cerevisiae to heterothallism is described which is applicable to genetically well behaved diploids, as well as to strains that sporulate poorly or produce few viable and mating-competent spores. The heterothallic (ho) allele was introduced into three widely used wine strains through spore x cell hybridization. The resultant hybrids were sporulated, and heterothallic segregants were isolated for use in successive backcrosses. Heterothallic progeny of opposite mating type and monosomic for chromosome III produced by sixth-backcross hybrids or their progeny were mated together to reconstruct heterothallic derivatives of the wine strain parents. A helpful prerequisite to the introduction of ho was genetic purification of the parental strains based on repeated cycles of sporulation, ascus dissection, and clonal selection. A positive selection to isolate laboratory-wine strain hybrids requiring no prior genetic alteration of the industrial strains, coupled with a partial selection to reduce the number of spore progeny needed to be screened to isolate heterothallic segregants of the proper genotype made the procedure valuable for genetically intractable strains. Trial grape juice fermentations indicated that introduction of ho had no deleterious effect on fermentation behavior. PMID- 16348172 TI - Partial Purification and Characterization of Two Hydrogenases from the Extreme Thermophile Methanococcus jannaschii. AB - F(420)-nonreactive and F(420)-reactive hydrogenases have been partially purified from Methanococcus jannaschii, an extremely thermophilic methanogen isolated from a submarine hydrothermal vent. The molecular weights of both hydrogenases were determined by native gradient electrophoresis in 5 to 27% polyacrylamide gels. The F(420)-nonreactive hydrogenase produced one major band (475 kilodaltons), whereas the F(420)-reactive hydrogenase produced two major bands (990 and 115 kilodaltons). The F(420)-nonreactive hydrogenase consisted of two subunits (43 and 31 kilodaltons), and the F(420)-reactive hydrogenase contained three subunits (48, 32, and 25 kilodaltons). Each hydrogenase was active at very high temperatures. Methyl viologen-reducing activity of the F(420)-nonreactive hydrogenase was maximal at 80 degrees C but was still detectable at 103 degrees C. The maximum activities of F(420)-reactive hydrogenase for F(420) and methyl viologen were measured at 80 and 90 degrees C, respectively. Low but measureable activity toward methyl viologen was repeatedly observed at 103 degrees C. Moreover, the half-life of the F(420)-nonreactive hydrogenase at 70 degrees C was over 9 h, and that of the F(420)-reactive enzyme was over 3 h. PMID- 16348173 TI - Adhesion of an Amylolytic Arthrobacter sp. to Starch-Containing Plastic Films. AB - Cells of the amylolytic bacterium KB-1 (thought to be an Arthrobacter sp.) adhered ( approximately 70%) to the surface of plastic films composed of starch poly (methylacrylate) graft copolymer (starch-PMA), but did not adhere (<10%) to films composed of polymethylacrylate (PMA), polyethylene (PE), carboxymethyl cellulose, or a mixture of PE plus poly (ethylene-coacrylic acid) (EAA), starch plus PE, or starch plus PE and EAA. About 30% of the cells adhered to gelatinized insoluble starch. Dithiothreitol (5 mM), EDTA (5 mM), and soluble starch (1%, wt/vol) had little effect on the adhesion of KB-1 cells to starch-PMA films. However, glutaraldehyde-fixed cells, azide-treated cells, and heat-killed cells did not bind to starch-PMA plastic, suggesting that the observed adhesion required cell viability. Culture supernatant from 5-day-old KB-1 cultures contained a proteolytic enzyme that inhibited cell adhesion to starch-PMA plastics. Trypsin-treated KB-1 cells also lost their ability to bind to starch PMA plastic. When washed free of trypsin and suspended in fresh medium, trypsin treated bacteria were able to recover adhesion activity in the absence, but not in the presence, of the protein synthesis inhibitor chloramphenicol. These results suggested that adhesion of KB-1 to starch-PMA plastic may be mediated by a cell surface protein. Although KB-1 bacteria bound to starch-PMA plastic, they did not appear to degrade starch in these films. Evidence of starch degradation was observed for starch-PE-EAA plastics, where <10% of the bacteria was bound, suggesting that cell adhesion may not be a prerequisite for degradation of some starch-containing plastics. PMID- 16348174 TI - Characterization of an endo-Acting Amylopullulanase from Thermoanaerobacter Strain B6A. AB - A thermoanaerobe (Thermoanaerobacter sp.) grown in TYE-starch (0.5%) medium at 60 degrees C produced both extra- and intracellular pullulanase (1.90 U/ml) and amylase (1.19 U/ml) activities. Both activities were produced at high levels on a variety of carbon sources. The temperature and pH optima for both pullulanase and amylase activities were 75 degrees C and pH 5.0, respectively. Both the enzyme activities were stable up to 70 degrees C (without substrate) and at pH 4.5 to 5.0. The half-lives of both enzyme activities were 5 h at 70 degrees C and 45 min at 75 degrees C. The enzyme activities did not show any metal ion activity, and both activities were inhibited by beta- and gamma-cyclodextrins but not by alpha cyclodextrin. A single amylolytic pullulanase responsible for both activities was purified to homogeneity by DEAE-Sepharose CL-6B column chromatography, gel filtration using high-pressure liquid chromatography, and pullulan-Sepharose affinity chromatography. It was a 450,000-molecular-weight glycoprotein composed of two equivalent subunits. The pullulanase cleaved pullulan in alpha1,6 linkages and produced multiple saccharides from cleavage of alpha-1,4 linkages in starch. The K(m)s for pullulan and soluble starch were 0.43 and 0.37 mg/ml, respectively. PMID- 16348175 TI - Batch tests to determine activity distribution and kinetic parameters for acetate utilization in expanded-bed anaerobic reactors. AB - Batch tests to measure maximum acetate utilization rates were used to determine the distribution of acetate utilizers in expanded-bed sand and expanded-bed granular activated carbon (GAC) reactors. The reactors were fed a mixture of acetate and 3-ethylphenol, and they contained the same predominant aceticlastic methanogen, Methanothrix sp. Batch tests were performed both on the entire reactor contents and with media removed from the reactors. Results indicated that activity was evenly distributed within the GAC reactors, whereas in the sand reactor a sludge blanket on top of the sand bed contained approximately 50% of the activity. The Monod half-velocity constant (K(s)) for the acetate-utilizing methanogens in two expanded-bed GAC reactors was searched for by combining steady state results with batch test data. All parameters necessary to develop a model with Monod kinetics were experimentally determined except for K(s). However, K(s) was a function of the effluent 3-ethylphenol concentration, and batch test results demonstrated that maximum acetate utilization rates were not a function of the effluent 3-ethylphenol concentration. Addition of a competitive inhibition term into the Monod expression predicted the dependence of K(s) on the effluent 3 ethylphenol concentration. A two-parameter search determined a K(s) of 8.99 mg of acetate per liter and a K(i) of 2.41 mg of 3-ethylphenol per liter. Model predictions were in agreement with experimental observations for all effluent 3 ethylphenol concentrations. Batch tests measured the activity for a specific substrate and determined the distribution of activity in the reactor. The use of steady-state data in conjunction with batch test results reduced the number of unknown kinetic parameters and thereby reduced the uncertainty in the results and the assumptions made. PMID- 16348176 TI - Production of the antibiotic phenazine-1-carboxylic Acid by fluorescent pseudomonas species in the rhizosphere of wheat. AB - Pseudomonas fluorescens 2-79 and P. aureofaciens 30-84 produce the antibiotic phenazine-1-carboxylic acid and suppress take-all, an important root disease of wheat caused by Gaeumannomyces graminis var. tritici. To determine whether the antibiotic is produced in situ, wheat seeds were treated with strain 2-79 or 30 84 or with phenazine-nonproducing mutants or were left untreated and then were sown in natural or steamed soil in the field or growth chamber. The antibiotic was isolated only from roots of wheat colonized by strain 2-79 or 30-84 in both growth chamber and field studies. No antibiotic was recovered from the roots of seedlings grown from seeds treated with phenazine-nonproducing mutants or left untreated. In natural soils, comparable amounts of antibiotic (27 to 43 ng/g of root with adhering soil) were recovered from roots colonized by strain 2-79 whether or not the pathogen was present. Roots of plants grown in steamed soil yielded larger bacterial populations and more antibiotic than roots from natural soils. In steamed and natural soils, roots from which the antibiotic was recovered had significantly less disease than roots with no antibiotic, indicating that suppression of take-all is related directly to the presence of the antibiotic in the rhizosphere. PMID- 16348177 TI - Utilization of Glycerol as a Hydrogen Acceptor by Lactobacillus reuteri: Purification of 1,3-Propanediol:NAD Oxidoreductase. AB - Lactobacillus reuteri utilizes exogenously added glycerol as a hydrogen acceptor during carbohydrate fermentations, resulting in higher growth rates and cell yields than those obtained during growth on carbohydrates alone. Glycerol is first converted to 3-hydroxypropionaldehyde by a coenzyme B(12)-dependent glycerol dehydratase and then reduced to 1,3-propanediol by an NAD -dependent oxidoreductase. The latter enzyme was purified and determined to have a molecular weight of 180,000; it is predicted to exist as a tetramer of identical 42,000 molecular-weight subunits. PMID- 16348178 TI - Metabolism of One-Carbon Compounds by the Ruminal Acetogen Syntrophococcus sucromutans. AB - Syntrophococcus sucromutans is the predominant species capable of O demethylation of methoxylated lignin monoaromatic derivatives in the rumen. The enzymatic characterization of this acetogen indicated that it uses the acetyl coenzyme A (Wood) pathway. Cell extracts possess all the enzymes of the tetrahydrofolate pathway, as well as carbon monoxide dehydrogenase, at levels similar to those of other acetogens using this pathway. However, formate dehydrogenase could not be detected in cell extracts, whether formate or a methoxyaromatic was used as electron acceptor for growth of the cells on cellobiose. Labeled bicarbonate, formate, [1-C] pyruvate, and chemically synthesized O-[methyl-C]vanillate were used to further investigate the catabolism of one-carbon (C(1)) compounds by using washed-cell preparations. The results were consistent with little or no contribution of formate dehydrogenase and pointed out some unique features. Conversion of formate to CO(2) was detected, but labeled formate predominantly labeled the methyl group of acetate. Labeled CO(2) readily exchanged with the carboxyl group of pyruvate but not with formate, and both labeled CO(2) and pyruvate predominantly labeled the carboxyl group of acetate. No CO(2) was formed from O demethylation of vanillate, and the acetate produced was position labeled in the methyl group. The fermentation pattern and specific activities of products indicated a complete synthesis of acetate from pyruvate and the methoxyl group of vanillate. PMID- 16348179 TI - Involvement of a Collar Structure in Polar Growth and Cell Division of Strain DCB 1. AB - Microscopic methods were used to investigate the unique collar structure of the gram-negative sulfate-reducing bacterium, strain DCB-1. Polar cell growth apparently occurred from the collar. When the daughter cell was approximately equal in length to the mother cell and the collar was thus centrally located, cell division occurred within the collar region. Division was by a novel mechanism which conserved the collar of the mother cell and gave rise to a new collar of the daughter cell. Cells of DCB-1 were also found to contain stacked internal membranes and glycogen bodies. PMID- 16348180 TI - Catabolism of tritiated thymidine by aquatic microbial communities and incorporation of tritium into RNA and protein. AB - The incorporation of tritiated thymidine by five microbial ecosystems and the distribution of tritium into DNA, RNA, and protein were determined. All microbial assemblages tested exhibited significant labeling of RNA and protein (i.e., nonspecific labeling), as determined by differential acid-base hydrolysis. Nonspecific labeling was greatest in sediment samples, for which >/=95% of the tritium was recovered with the RNA and protein fractions. The percentage of tritium recovered in the DNA fraction ranged from 15 to 38% of the total labeled macromolecules recovered. Nonspecific labeling was independent of both incubation time and thymidine concentration over very wide ranges. Four different RNA hydrolysis reagents (KOH, NaOH, piperidine, and enzymes) solubilized tritium from cold trichloroacetic acid precipitates. High-pressure liquid chromatography separation of piperidine hydrolysates followed by measurement of isolated monophosphates confirmed the labeling of RNA and indicated that tritium was recovered primarily in CMP and AMP residues. We also evaluated the specificity of [2-H]adenine incorporation into adenylate residues in both RNA and DNA in parallel with the [H]thymidine experiments and compared the degree of nonspecific labeling by [H]adenine with that derived from [H]thymidine. Rapid catabolism of tritiated thymidine was evaluated by determining the disappearance of tritiated thymidine from the incubation medium and the appearance of degradation products by high-pressure liquid chromatography separation of the cell-free medium. Degradation product formation, including that of both volatile and nonvolatile compounds, was much greater than the rate of incorporation of tritium into stable macromolecules. The standard degradation pathway for thymidine coupled with utilization of Krebs cycle intermediates for the biosynthesis of amino acids, purines, and pyrimidines readily accounts for the observed nonspecific labeling in environmental samples. PMID- 16348181 TI - Role of Polysulfides in Reduction of Elemental Sulfur by the Hyperthermophilic Archaebacterium Pyrococcus furiosus. AB - Polysulfides formed through the breakdown of elemental sulfur or other sulfur compounds were found to be reduced to H(2)S by the hyperthermophilic archaebacterium Pyrococcus furiosus during growth. Metabolism of polysulfides by the organism was dissimilatory, as no incorporation of S-labeled elemental sulfur was detected. However, [S]cysteine and [S]methionine were incorporated into cellular protein. Contact between the organism and elemental sulfur is not necessary for metabolism. The sulfide generated from metabolic reduction of polysulfides dissociates to a strong nucleophile, HS, which in turn opens up the S(8) elemental sulfur ring. In addition to H(2)S, P. furiosus cultures produced methyl mercaptan in a growth-associated fashion. PMID- 16348182 TI - DNA probes show genetic variation in cyanobacterial symbionts of the azolla fern and a closer relationship to free-living nostoc strains than to free-living anabaena strains. AB - Twenty-two isolates of Anabaena azollae derived from seven Azolla species from various geographic and ecological sources were characterized by DNA-DNA hybridization. Cloned DNA fragments derived from the genomic sequences of three different A. azollae isolates were used to detect restriction fragment length polymorphism among all symbiotic anabaenas. DNA clones were radiolabeled and hybridized against southern blot transfers of genomic DNAs of different isolates of A. azollae digested with restriction endonucleases. Eight DNA probes were selected to identify the Anabaena strains tested. Two were strain specific and hybridized only to A. azollae strains isolated from Azolla microphylla or Azolla caroliniana. One DNA probe was section specific (hybridized only to anabaenas isolated from Azolla ferns representing the section Euazolla), and five other probes gave finer discrimination among anabaenas representing various ecotypes of Azolla species. These cloned genomic DNA probes identified 11 different genotypes of A. azollae isolates. These included three endosymbiotic genotypes within Azolla filiculoides species and two genotypes within both A. caroliniana and Azolla pinnata endosymbionts. Although we were not able to discriminate among anabaenas extracted from different ecotypes of Azolla nilotica, Azolla mexicina, Azolla rubra and Azolla microphylla species, each of the endosymbionts was easily identified as a unique genotype. When total DNA isolated from free-living Anabaena sp. strain PCC7120 was screened, none of the genomic DNA probes gave detectable positive hybridization. Total DNA of Nostoc cycas PCC7422 hybridized with six of eight genomic DNA fragments. These data imply that the dominant symbiotic organism in association with Azolla spp. is more closely related to Nostoc spp. than to free-living Anabaena spp. PMID- 16348183 TI - Evaluation and reliability of a simplified method for identification of food borne yeasts. AB - A simplified identification key described by Deak and Beuchat (T. Deak and L. R. Beuchat, J. Food Prot. 50:243-264, 1987) and the computer method of Barnett et al. (J. A. Barnett, R. W. Payne, and D. Yarrow, Yeast Identification Program, 1985) were used to identify 12 reference strains and 382 yeasts isolated from cultured milk products. Because the simplified key failed to account for species variability with regard to physiological, morphological, and sexual reproduction characteristics, poor agreement of the identification results was obtained. A reevaluation of the basic theoretical assumptions of the simplified key only confirmed the practical results and indicates that this identification method is unsatisfactory. PMID- 16348184 TI - Carbon- and Nitrogen-to-Volume Ratios of Bacterioplankton Grown under Different Nutritional Conditions. AB - Carbon- and nitrogen-to-volume (C/V and N/V) ratios were determined for freshwater bacterial assemblages grown in lake water filtrate or in water enriched with nutrients (aqueous extract of lake seston, glucose, arginine, phosphate, or ammonium). Biovolume was measured by epifluorescence microphotography, and carbon and nitrogen biomasses were measured with a CHN analyzer. Despite large variations of nutritional conditions (i.e., the composition and concentration of the dissolved organic carbon) and different mean cell sizes of the bacterial assemblage (0.17 to 1.8 mum per cell), the C/V, N/V, and carbon-to-nitrogen weight ratios varied little (C/V ratio, 0.14 pg of C per mum [standard deviation, 0.057; n = 15]; N/V ratio, 0.027 pg of N per mum [standard deviation; 0.011, n = 15]; carbon-to-nitrogen weight ratio, 5.6 [standard deviation, 2.2, n = 15]). An average C/V ratio of 0.12 pg of C per mum that was derived from natural and cultured bacterial assemblages is proposed as an appropriate conversion factor for estimation of the biomass of freshwater bacteria. PMID- 16348185 TI - Adhesive properties of a symbiotic bacterium from a wood-boring marine shipworm. AB - Adhesive properties of a cellulolytic, nitrogen-fixing bacterium isolated from a marine shipworm by Waterbury et al. (J. B. Waterbury, C. B. Calloway, and R. D. Turner, Science 221:1401-1403, 1983) are described. S-labeled cells of the shipworm bacterium bound preferentially to Whatman no. 1 cellulose filter paper, compared with its binding to other cellulose substrata or substrata lacking cellulose. The ability of the bacteria to bind to Whatman no. 1 filter paper was significantly reduced by glutaraldehyde or heat treatment of cells. Pretreatment of cells with azide, valinomycin, gramicidin-D, bis-hexafluoroacetylacetone (1799), or carbonyl cyanide-p-trifluoromethoxyphenylhydrazone inhibited adhesion activity. Cells pretreated with pronase or trypsin also exhibited reduced binding activity, but chymotrypsin and peptidase had no effect on adhesion activity. Cellodextrins and methyl cellulose 15 inhibited the adhesion of shipworm bacteria to filter paper, whereas glucose, cellobiose, and soluble carboxymethyl cellulose had no significant effect. The divalent cation chelators EDTA and EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N'N'-tetraacetic acid] had little or no effect on adhesive properties of shipworm bacteria. Also, preabsorbing the substratum with extracellular endoglucanase isolated from the shipworm bacterium or 1% bovine serum albumin had no apparent effect on bacterial binding. Low concentrations (0.01%) of sodium dodecyl sulfate solubilized a fraction from whole cells, which appeared to be involved in cellular binding activity. After removal of sodium dodecyl sulfate, several proteins in this fraction associated with intact cells. These cells exhibited up to 50% enhanced binding to filter paper in comparison to cells which had not been exposed to the sodium dodecyl sulfate-solubilized fraction. PMID- 16348186 TI - Strain-Specific Inhibition of nod Gene Induction in Bradyrhizobium japonicum by Flavonoid Compounds. AB - A broad-host-range plasmid, pEA2-21, containing a Bradyrhizobium japonicum nodABC'-'lacZ translational fusion was used to identify strain-specific inhibitors of the genes required for soybean nodulation, the common nod genes. The responses of type strains of B. japonicum serogroups USDA 110, USDA 123, USDA 127, USDA 129, USDA 122, and USDA 138 to nod gene inhibitors were compared. Few compounds inhibited nod gene expression in B. japonicum USDA 110. In contrast, nod gene expression in strains belonging to several other serogroups was inhibited by most of the flavonoids tested. However, the application of two of these strain-specific compounds, chrysin and naringenin, had little effect on the pattern of competition between indigenous and inoculum strains of B. japonicum in greenhouse and field trials. Preliminary studies with radiolabeled chrysin and naringenin suggest that the different responses to nod gene inhibitors may be partly due to the degree to which plant flavonoids can be metabolized by each strain. PMID- 16348187 TI - Oligonucleotide Probes That Hybridize with rRNA as a Tool To Study Frankia Strains in Root Nodules. AB - Oligonucleotide probes that hybridize with specific sequences in variable regions of the 16S rRNA of the nitrogen-fixing actinomycete Frankia were used for the identification of Frankia strains in nodules. Frankia cells were released from plant tissue by grinding glutaraldehyde-fixed root nodules in guanidine hydrochloride solution. rRNA was obtained after sonication, precipitation with ethanol, and purification by phenolchloroform extraction. Degradation of rRNA, evident in Northern blots, did not affect hybridization with the oligonucleotides. Nodules of about 1 mg (fresh weight) provided sufficient rRNA for reliable detection of the Frankia strain. The utility of this rRNA extraction method was tested in a competition experiment between two effective Frankia strains on cloned Alnus glutinosa plants. PMID- 16348188 TI - Coupled metabolic and photolytic pathway for degradation of pyridinedicarboxylic acids, especially dipicolinic Acid. AB - Three isomers of pyridinedicarboxylic acid (PDCA) (2,3-, 2,5-, and 2,6-PDCA) were partially oxidized by marine bacteria when grown aerobically on the corresponding phthalate analogs. The metabolites, unlike the parent PDCAs, absorbed light in the solar actinic range (wavelengths greater than 300 nm) and were readily degraded in sunlight. The principal product from 2,6-PDCA (dipicolinic acid) metabolism was extracted from a culture fluid, purified by column chromatography, and analyzed by UV-visible, infrared, and C nuclear magnetic resonance spectroscopy. The compound was identified as 2,3-dihydroxypicolinic acid (2,3 DHPA). 2,3-DHPA was photolyzed in aqueous solution (pH 8.0) with a half-life of 100 min. Eight photoproducts, three of which were photolabile, were detected by high-performance liquid chromatography. Ammonia was also photoproduced from 2,3 DHPA. Analysis of the photoproducts by UV-visible spectroscopy and by high performance liquid chromatography of 2,4-dinitrophenylhydrazones indicated that the products were conjugated carbonyls and carboxylic acids. Six of the photoproducts were readily consumed by bacterial strain CC9M. In illuminated aquatic environments, coupled bio- and photodegradative mechanisms probably contribute to the degradation of PDCAs. PMID- 16348189 TI - Effect of Indoleacetic Acid and Related Indoles on Lactobacillus sp. Strain 11201 Growth, Indoleacetic Acid Catabolism, and 3-Methylindole Formation. AB - A study was conducted to determine the activity of the 3-methylindole (3MI) forming enzyme in Lactobacillus sp. strain 11201. Cells were incubated anaerobically with 17 different indolic and aromatic compounds. Indoleacetic acid (IAA), 5-hydroxyindoleacetic acid, 5-methoxy-3-indoleacetic acid, indole-3 pyruvate, or indole-3-propionic acid induced 3MI-forming activity. The highest total enzyme activity induced by IAA was observed in cells incubated with an initial concentration of 1.14 mM IAA. Peak activity of the 3MI-forming enzyme occurred 4 h after bacteria were incubated with either 0.114 or 1.14 mM IAA. Enzyme activity peaked earlier (2 h) and disappeared more rapidly at 5.7 mM IAA than at other concentrations of IAA. The effects of IAA and 3MI on the growth of Lactobacillus sp. strain 11201 and formation of 3MI from IAA also were determined. Bacterial growth and 3MI formation from IAA were reduced in medium containing exogenous 3MI. The growth depression observed in medium containing 5.7 mM IAA appears to be due to the toxicity of 3MI rather than IAA. The formation of 3MI in this ruminal Lactobacillus sp. is mediated by an inducible enzyme, and as 3MI accumulates, bacterial growth and rates of 3MI formation from IAA are reduced. PMID- 16348190 TI - Viruses as partners in spring bloom microbial trophodynamics. AB - Population sizes of algae, bacteria, heterotrophic flagellates, and viruses were observed through the 1989 spring diatom bloom in Raunefjorden in western Norway. The culmination of the diatom bloom was followed by a peak in the concentration of bacteria and an increase in the concentration of heterotrophic flagellates, a pattern consistent with the concept of a food chain from photosynthetically produced organic material, through bacteria, to bacterivorous flagellates. The concentration of viruses varied through the spring bloom from 5 x 10 in the prebloom situation to a maximum of 1.3 x 10 viruses ml 1 week after the peak of the diatom bloom. Coinciding with the collapse in the diatom bloom, a succession of bacteria and viruses was observed in the mucous layer surrounding dead or senescent diatoms, with an estimated maximum of 23% of the total virus population attached to the diatoms. The dynamic behavior observed for the virus population rules out the possibility that it is dominated by inactive species, and the viruses are suggested to be active members of the microbial food web as agents causing lysis in parts of the bacterial population, diverting part of the bacterial production from the predatory food chain. PMID- 16348191 TI - New method for extraction of streptomycete spores from soil and application to the study of lysogeny in sterile amended and nonsterile soil. AB - A new method for the isolation and enumeration of streptomycete spores from soil was developed. This method makes use of a cation-exchange resin to disperse soil particles. It allowed the detection of 10 spores in 100 g of sterile soil, while ca. 10 could be accurately enumerated in 100 g. This method was applied to studying the fate of a marked actinophage in soil. In sterile amended and nonsterile soil, relatively high numbers of actinophages were only found during the first few days of the experiment when the host streptomycete was in the mycelial form. Later, after sporulation, lysogens could be detected in sterile amended soil and could still be found 60 days after inoculation. Although no lysogens were found in nonsterile soil, the introduced phage could still be detected in the free state after 60 days, albeit at a low titer. PMID- 16348192 TI - Growth and survival of streptomycete inoculants and extent of plasmid transfer in sterile and nonsterile soil. AB - The growth and survival of strains of Streptomyces lividans and S. violaceolatus in sterile and nonsterile soil was investigated by using inoculated soil microcosms run as batch systems. It was evident that, after an initial short mycelial growth phase of 2 to 3 days, sporulation occurred and inoculants survived as spores. The transfer of a high-copy-number, self-transmissible plasmid, pIJ673, was detected by using intra- and interspecific crosses. The initial detection of transconjugants correlated with the development of the mycelial state of the inoculants (as confirmed by scanning electron microscopy) after 2 days of incubation. Subsequent spread of the plasmid was attributed to spread within existing mycelium followed by sporulation. In natural soil, inoculant numbers remained constant or declined, but plasmid transfer was readily detected. PMID- 16348193 TI - Particulate DNA in smoker fluids: evidence for existence of microbial populations in hot hydrothermal systems. AB - As part of an interdisciplinary study of hydrothermal vents on the Endeavour Segment of the Juan de Fuca Ridge, we used the submersible ALVIN to collect 57 fluid samples in titanium syringes and Go Flo Niskin bottles from 17 different hot vents (smokers and flanges) and their environs for the purpose of extracting particulate DNA. The relative purity of the vent fluids collected was determined by Mg content as an indicator of seawater entrainment. Particulate material concentrated from these samples was lysed enzymatically (enz) and by a combination of enzyme and French press treatment (fp). Concentrations of partially purified DNA recovered from these lysates were determined spectrofluorometrically by using the dye Hoechst 33258. Ambient seawater surrounding the vents was found to contain low DNA concentrations, 0.18 to 0.32 ng of DNA per ml (n = 4; mean(enz) = 0.23 +/- 0.05; mean(fp) = 0.26 +/- 0.05), while low-temperature vent samples yielded significantly higher concentrations of 0.37 to 2.12 ng of DNA per ml (n = 4; mean(enz) = 0.97 +/- 0.68; mean(fp) = 1.05 +/- 0.54). Although DNA recovery values from superheated (210 to 345 degrees C) flange samples (mean(enz) = 0.14 +/- 0.10; mean(fp) = 0.12 +/- 0.14) were not significantly different from ambient seawater values, most of the superheated (174 to 357 degrees C) smoker fluid samples contained particulate DNA in concentrations too high to be attributable to entrained seawater. Detailed sampling at one smoker site demonstrated not only the existence of significant levels of particulate DNA in the superheated smoker fluids but also the presence of an elevated microbial population in the buoyant plume 20 to 100 m above the smoker. These results underscore the heterogeneity of smoker environments within a given hydrothermal vent field and indicate that microorganisms exist in some superheated fluids. PMID- 16348194 TI - Specificity of octopine uptake by Rhizobium and pseudomonas strains. AB - The octopine-utilizing strain Agrobacterium tumefaciens B6S3 and three nonagrobacteria which had the capacity to utilize this opine were compared for octopine uptake. The characteristics of uptake by Rhizobium meliloti A3 and strain B6S3 were similar. In both bacteria, uptake activity was inducible by octopine and by the related opine octopinic acid, and competition assays showed that these two opine substrates were accepted by the same uptake system with an equivalent affinity. Cells of Pseudomonas putida 203 accumulated octopine against a concentration gradient, and this activity was induced specifically by octopine. While strain 203 did not utilize octopinic acid, a spontaneous mutant with a combined capacity for octopine and octopinic acid utilization was obtained. Both opines induced octopine uptake by this mutant, but octopinic acid was not a substrate for the induced system. Thus, the Pseudomonas uptake system exhibited a different specificity for octopine than the corresponding Agrobacterium system. The nonfluorescent pseudomonad GU187j, which utilized the three related opines octopine, octopinic acid, and nopaline, was constitutive for octopine uptake. Strain GU187j possessed a system which accepted these three opines, but not arginine or ornithine, with a similar affinity. PMID- 16348195 TI - Nonradioactive screening method for isolation of disease-specific probes to diagnose plant diseases caused by mycoplasmalike organisms. AB - DNA fragments of tomato big bud (BB) mycoplasmalike organism (MLO) in diseased periwinkle plants (Catharanthus roseus L.) were cloned to pSP6 plasmid vectors and amplified in Escherichia coli JM83. A nonradioactive method was developed and used to screen for MLO-specific recombinants. Cloned DNA probes were prepared by nick translation of the MLO recombinant plasmids by using biotinylated nucleotides. The probes all hybridized with nucleic acid from BB MLO-infected, but not healthy, plants. Results from dot hybridization analyses indicated that several MLOs, e.g., those of Italian tomato big bud, periwinkle little leaf, and clover phyllody, are closely related to BB MLO. The Maryland strain of aster yellows and maize bushy stunt MLOs are also related to BB MLO. Among the remaining MLOs used in this study, Vinca virescence and elm yellows MLOs may be very distantly related, if at all, to BB MLO. Potato witches' broom, clover proliferation, ash yellows, western X, and Canada X MLOs are distantly related to BB MLO. Southern hybridization analyses revealed that BB MLO contains extrachromosomal DNA that shares sequence homologies with extrachromosomal DNAs from aster yellows and periwinkle little leaf MLOs. PMID- 16348196 TI - DNA probe-mediated detection of resistant bacteria from soils highly polluted by heavy metals. AB - Alcaligenes eutrophus CH34 DNA fragments encoding resistance to Cd, Co, Zn (czc), or Hg (merA) were cloned and used as probes in colony hybridization procedures with bacteria isolated from polluted environments such as a zinc factory area (desertified because of the toxic effects of zinc contamination) and from sediments from factories of nonferrous metallurgy in Belgium and mine areas in Zaire. From the different soil samples, strains could be isolated and hybridized with the czc probe (resistance to Cd, Co, and Zn from plasmid pMOL30). Percentages of CFU isolated on nonselective plates which hybridized with the czc and the mercury resistance probes were, respectively, 25 and 0% for the zinc desert, 15 to 20 and 10 to 20% for the two Belgian factories, and 40 and 40% for the Likasi mine area. Most of these strains also carried two large plasmids of about the same size as those of A. eutrophus CH34 and shared many phenotypic traits with this strain. These findings indicated a certain correlation between the heavy-metal content in contaminated soils and the presence of heavy-metal resistant megaplasmid-bearing A. eutrophus strains. PMID- 16348197 TI - Transconjugation between Bacteria in the Digestive Tract of the Cutworm Peridroma saucia. AB - Transconjugants arising from transfer of plasmid R388::Tn1721 between donor and recipient strains of Enterobacter cloacae were detected in samples from the digestive tracts and fecal pellets of variegated cutworms (Peridroma saucia). PMID- 16348198 TI - Improved Method for the Isolation of Biosurfactant Glycolipids from Rhodococcus sp. Strain H13A. AB - An improved method for the isolation of the biosurfactant glycolipids from Rhodococcus sp. strain H13A by using XM 50 diafiltration and isopropanol precipitation was devised. This procedure was advantageous since it removes protein coisolated when the glycolipids are obtained by organic extraction and silicic acid chromatography. The protein apparently does not contribute any biosurfactant characteristics to the glycolipids. The deacylated glycolipid backbone included only a disaccharide. PMID- 16348199 TI - Isolation, identification, and characterization of a feather-degrading bacterium. AB - A feather-degrading culture was enriched with isolates from a poultry waste digestor and adapted to grow with feathers as its primary source of carbon, sulfur, and energy. Subsequently, a feather-hydrolytic, endospore-forming, motile, rod-shaped bacterium was isolated from the feather-degrading culture. The organism was Gram stain variable and catalase positive and demonstrated facultative growth at thermophilic temperatures. The optimum rate of growth in nutrient broth occurred at 45 to 50 degrees C and at pH 7.5. Electron microscopy of the isolate showed internal crystals. The microorganism was identified as Bacillus licheniformis PWD-1. Growth on hammer-milled-feather medium of various substrate concentrations was determined by plate colony count. Maximum growth (approximately 10 cells per ml) at 50 degrees C occurred 5 days postinoculation on 1% feather substrate. Feather hydrolysis was evidenced as free amino acids produced in the medium. The most efficient conditions for feather fermentation occurred during the incubation of 1 part feathers to 2 parts B. licheniformis PWD 1 culture (10 cells per ml) for 6 days at 50 degrees C. These data indicate a potential biotechnique for degradation and utilization of feather keratin. PMID- 16348200 TI - Simple, sensitive zymogram technique for detection of xylanase activity in polyacrylamide gels. AB - A method capable of detecting as little as 0.11 U of xylanase activity in polyacrylamide gels was developed. The method entails incubation of protein gels in contact with substrate gels containing unmodified xylan, followed by immersion of substrate gels in 95% ethanol. Resulting zymograms contain transparent bands corresponding to enzymatic activity against an opaque background. PMID- 16348201 TI - Fractionation of the beta-Linked Glucans of Bradyrhizobium japonicum and Their Response to Osmotic Potential. AB - Bradyrhizobium japonicum USDA 110 synthesized both extracellular and periplasmic polysaccharides when grown on mannitol minimal medium. The extracellular polysaccharides were separated into a high-molecular-weight acidic capsular extracellular polysaccharide fraction (90% of total hexose) and three lower molecular-weight glucan fractions by liquid chromatography. Periplasmic glucans, extracted from washed cells with 1% trichloroacetic acid, gave a similar pattern on liquid chromatography. Linkage analysis of the major periplasmic glucan fractions demonstrated mainly 6-linked glucose (63 to 68%), along with some 3,6- (8 to 18%), 3- (9 to 11%), and terminal (7 to 8%) linkages. The glucose residues were beta-linked as shown by H-nuclear magnetic resonance analysis. Glucan synthesis by B. japonicum cells grown on mannitol medium with 0 to 350 mM fructose as osmolyte was measured. Fructose at 150 mM or higher inhibited synthesis of periplasmic and extracellular 3- and 6-linked glucans but had no effect on the synthesis of capsular acidic extracellular polysaccharides. PMID- 16348202 TI - Photoadaptation and Protection against Active Forms of Oxygen in the Symbiotic Procaryote Prochloron sp. and Its Ascidian Host. AB - Superoxide dismutase, ascorbate peroxidase, and catalase activities were studied in the symbiotic photosynthetic procaryote Prochloron sp. and its ascidian host Lissoclinum patella. The protein-specific activities of these antioxidant enzymes in the Prochloron sp. and L. patella collected at different depths from the Great Barrier Reef, Australia, were directly proportional to irradiance, whereas the pigment concentrations in the Prochloron sp. were inversely proportional to irradiance. The presence of a cyanide-sensitive superoxide dismutase, presumably a Cu-Zn metalloprotein, in the Prochloron sp. extends the possible phylogenetic distribution of this protein. The concentration of UV-absorbing mycosporine-like amino acids is inversely proportional to irradiance in both the host and symbiont, suggesting that these compounds may not provide sufficient protection against UV radiation in high-irradiance environments. The significant differences in the specific activities of these antioxidant enzymes, cellular photosynthetic pigment concentrations, and UV-absorbing compounds from high- and low-irradiance habitats constitute an adaptive response to different photic environments. These photoadaptive responses are essential to prevent inhibition of photosynthesis by high fluxes of visible and UV radiation. PMID- 16348203 TI - Supplemental substrate enhancement of 2,4-dinitrophenol mineralization by a bacterial consortium. AB - A Janthinobacterium sp. and an actinomycete, both capable of mineralizing 2,4 dinitrophenol (DNP), were used to construct a consortium to mineralize DNP in nonaxenic bench-scale sequencing batch reactors (SBRs). Average K(m) values for DNP mineralization by pure cultures of the Janthinobacterium sp. and the actinomycete were 0.01 and 0.13 mug/ml, respectively, and the average maximum specific growth rate (mu(max)) values for them were 0.06 and 0.23/h, respectively. In the presence of NH(4)Cl, nitrite accumulation in pure culture experiments and in the SBRs was stoichiometric to initial DNP concentration and the addition of nitrogen enhanced DNP mineralization in the SBRs. Mineralization of 10 mug of DNP per ml was further enhanced in SBRs by the addition of glucose at concentrations of 100 and 500 mug/ml but not at 10 mug/ml. Possible mechanisms for this enhanced DNP mineralization in SBRs were suggested by kinetic analyses and biomass measurements. Average mu(max) values for DNP mineralization in the presence of 0, 10, 100, and 500 mug of glucose per ml were 0.33, 0.13, 0.42, and 0.59/h, respectively. In addition, there was greater standing biomass in reactors amended with glucose. At steady-state operation, all SBRs contained heterogeneous microbial communities but only one organism, an actinomycete, that was capable of mineralizing DNP. This research demonstrates the usefulness of supplemental substrates for enhancing the degradation of toxic chemicals in bioreactors that contain heterogeneous microbial communities. PMID- 16348204 TI - Hydrocarbon mineralization in sediments and plasmid incidence in sediment bacteria from the campeche bank. AB - Rates of degradation of radiolabeled hydrocarbons and incidence of bacterial plasmid DNA were investigated in sediment samples collected from the Campeche Bank, Gulf of Mexico, site of an offshore oil field containing several petroleum platforms. Overall rates of mineralization of [C]hexadecane and [C]phenanthrene measured for sediments were negligible; <1% of the substrate was converted to CO(2) in all cases. Low mineralization rates are ascribed to nutrient limitations and to lack of adaptation by microbial communities to hydrocarbon contaminants. Plasmid frequency data for sediment bacteria similarly showed no correlation with proximity to the oil field, but, instead, showed correlation with water column depth at each sampling site. Significant differences between sites were observed for proportion of isolates carrying single or multiple plasmids and mean number of plasmids per isolate, each of which increased as a function of depth. PMID- 16348206 TI - Liquid-to-Gas Mass Transfer in Anaerobic Processes: Inevitable Transfer Limitations of Methane and Hydrogen in the Biomethanation Process. AB - Liquid-to-gas mass transfer in anaerobic processes was investigated theoretically and experimentally. By using the classical definition of k(L)a, the global volumetric mass transfer coefficient, theoretical development of mass balances in such processes demonstrates that the mass transfer of highly soluble gases is not limited in the usual conditions occurring in anaerobic fermentors (low-intensity mixing). Conversely, the limitation is important for poorly soluble gases, such as methane and hydrogen. The latter could be overconcentrated to as much as 80 times the value at thermodynamic equilibrium. Such overconcentrations bring into question the biological interpretations that have been deduced solely from gaseous measurements. Experimental results obtained in three different methanogenic reactors for a wide range of conditions of mixing and gas production confirmed the general existence of low mass transfer coefficients and consequently of large overconcentrations of dissolved methane and hydrogen (up to 12 and 70 times the equilibrium values, respectively). Hydrogen mass transfer coefficients were obtained from the direct measurements of dissolved and gaseous concentrations, while carbon dioxide coefficients were calculated from gas phase composition and calculation of related dissolved concentration. Methane transfer coefficients were based on calculations from the carbon dioxide coefficients. From mass balances performed on a gas bubble during its simulated growth and ascent to the surface of the liquid, the methane and carbon dioxide contents in the gas bubble appeared to be controlled by the bubble growth process, while the bubble ascent was largely responsible for a slight enrichment in hydrogen. PMID- 16348205 TI - Ferrous Iron and Sulfur Oxidation and Ferric Iron Reduction Activities of Thiobacillus ferrooxidans Are Affected by Growth on Ferrous Iron, Sulfur, or a Sulfide Ore. AB - Eight strains of Thiobacillus ferrooxidans (laboratory strains Tf-1 [= ATCC 13661] and Tf-2 [= ATCC 19859] and mine isolates SM-1, SM-2, SM-3, SM-4, SM-5, and SM-8) and three strains of Thiobacillus thiooxidans (laboratory strain Tt [= ATCC 8085] and mine isolates SM-6 and SM-7) were grown on ferrous iron (Fe), elemental sulfur (S), or sulfide ore (Fe, Cu, and Zn). The cells were studied for their aerobic Fe - and S-oxidizing activities (O(2) consumption) and anaerobic S oxidizing activity with ferric iron (Fe) (Fe formation). Fe-grown T. ferrooxidans cells oxidized S aerobically at a rate of 2 to 4% of the Fe oxidation rate. The rate of anaerobic S oxidation with Fe was equal to the aerobic oxidation rate in SM-1, SM-3, SM-4, and SM-5, but was only one-half or less that in Tf-1, Tf-2, SM 2, and SM-8. Transition from growth on Fe to that on S produced cells with relatively undiminished Fe oxidation activities and increased S oxidation (both aerobic and anaerobic) activities in Tf-2, SM-4, and SM-5, whereas it produced cells with dramatically reduced Fe oxidation and anaerobic S oxidation activities in Tf-1, SM-1, SM-2, SM-3, and SM-8. Growth on ore 1 of metal-leaching Fe-grown strains and on ore 2 of all Fe-grown strains resulted in very high yields of cells with high Fe and S oxidation (both aerobic and anaerobic) activities with similar ratios of various activities. Sulfur-grown Tf-2, SM-1, SM-4, SM-6, SM-7, and SM-8 cultures leached metals from ore 3, and Tf-2 and SM-4 cells recovered showed activity ratios similar to those of other ore-grown cells. It is concluded that all the T. ferrooxidans strains studied have the ability to produce cells with Fe and S oxidation and Fe reduction activities, but their levels are influenced by growth substrates and strain differences. PMID- 16348207 TI - Genes Responsible for Size Reduction of Marine Vibrios during Starvation Are Located on the Chromosome. AB - In a survey of 21 marine Vibrio spp., all responded to nutrient deprivation by undergoing a reduction in size (dwarfing). However, only 43% of these strains possessed one or more plasmids, suggesting that the genes responsible for dwarfing were located on the chromosome rather than on the plasmids. This conclusion was confirmed by the observation that fragmentation and subsequent size reduction occurred in three strains from which the plasmids had been removed by curing. The cured strains lost certain characteristics, such as resistance to some heavy metals and antibiotics, that were restored when the plasmids were reintroduced by either transformation or electroporation. PMID- 16348208 TI - Characterization of rhizobacteria associated with weed seedlings. AB - Rhizobacteria were isolated from seedlings of seven economically important weeds and characterized for potential phytopathogenicity, effects on seedling growth, and antibiosis to assess the possibility of developing deleterious rhizobacteria as biological control agents. The abundance and composition of rhizobacteria varied among the different weed species. For example, fluorescent pseudomonads represented from 11 to 42% of the total rhizobacterial populations from jimsonweed and lambsquarters, respectively. Other bacteria frequently isolated were nonfluorescent pseudomonads, Erwinia herbicola, Alcaligenes spp., and Flavobacterium spp. Only 18% of all isolates were potentially phytopathogenic, based on an Escherichia coli indicator bioassay. However, the proportion of isolates that inhibited growth in seedling assays ranged from 35 to 65% depending on the weed host. Antibiosis was most prevalent among isolates of fluorescent Pseudomonas spp., the activity of which was due to siderophore production in over 75% of these isolates. Overall, rhizobacterial isolates exhibited a complex array of properties that were inconsistent with accepted definitions for plant growth promoting and deleterious rhizobacteria. It is suggested that for development of effective biological control agents for weed control, deleterious rhizobacteria must be screened directly on host seedlings and must possess several properties including high colonizing ability, specific phytotoxin production, and resistance or tolerance to antibiotics produced by other rhizosphere microorganisms, and they must either synthesize or utilize other bacterial siderophores. PMID- 16348209 TI - Properties of 2,3-Butanediol Dehydrogenases from Lactococcus lactis subsp. lactis in Relation to Citrate Fermentation. AB - Two 2,3-butanediol dehydrogenases (enzymes 1 and 2; molecular weight of each, 170,000) have been partially purified from Lactococcus lactis subsp. lactis (Streptococcus diacetylactis) D10 and shown to have reductase activity with either diacetyl or acetoin as the substrate. However, the reductase activity with 10 mM diacetyl was far greater for both enzymes (7.0- and 4.7-fold for enzymes 1 and 2, respectively) than with 10 mM acetoin as the substrate. In contrast, when acetoin and diacetyl were present together, acetoin was the preferred substrate for both enzymes, with enzyme 1 showing the more marked preference for acetoin. meso-2,3-Butanediol was the only isomeric product, with enzyme 1 independent of the substrate combinations. For enzyme 2, both the meso and optical isomers of 2,3-butanediol were formed with acetoin as the substrate, but only the optical isomers were produced with diacetyl as the substrate. With batch cultures of strain D10 at or near the point of citrate exhaustion, the main isomers of 2,3 butanediol present were the optical forms. If the pH was sufficiently high (>pH 5), acetoin reduction occurred over time and was followed by diacetyl reduction, and meso-2,3-butanediol became the predominant isomer. Interconversion of the optical isomers into the meso isomer did occur. The properties of 2,3-butanediol dehydrogenases are consistent with diacetyl and acetoin removal and the appearance of the isomers of 2,3-butanediol. PMID- 16348210 TI - Bacterioplankton growth on fractions of dissolved organic carbon of different molecular weights from humic and clear waters. AB - The ability of fractions of dissolved organic carbon (DOC) of different molecular weights (MW) to support bacterial growth was studied in batch culture experiments. Natural pelagic bacteria were inoculated into particle-free (0.2-mum filtered) water, taken from 10 oligotrophic lakes of differing humic content, and either used without further treatments or ultrafiltered to remove DOC of >10,000 MW or >1,000 MW. Stationaryphase abundance of bacteria in the cultures was used as an estimate of bacterial carrying capacity. High-MW DOC (>10,000) comprised an increasing fraction of total DOC with increasing total DOC and humic content of the lakes. High-MW DOC was generally more available to bacteria (i.e., more bacteria were produced per unit of organic carbon initially present) than low-MW (<10,000) DOC. The availability to bacteria of this high-MW DOC decreased with increasing humic content. However, although less available in humic lakes than in clearwater lakes, the higher abundance of high-MW DOC made it quantitatively more important as a bacterial substrate; i.e., a larger fraction of the total bacterial yield of the cultures was due to high-MW DOC compounds in humic lakes than in clearwater lakes. On the average, 48% of bacterial growth occurred at the expense of DOC of <10,000 MW. DOC of <1,000 MW was responsible for an average of 22% of bacterial growth, with no significant correlation to humic content and DOC concentration of lakes. The DOC which supports bacterial growth, as well as the total DOC, is of different quality in humic and clearwater lakes. PMID- 16348211 TI - Effect of cetylpyridinium chloride on microbial adhesion to hexadecane and polystyrene. AB - Microbial adhesion at the oil-water interface is a subject of both basic interest (e.g., as a technique for the measurement of hydrophobicity) and applied interest (e.g., for use in two-phase oil-water mouthwashes for the desorption of oral microorganisms). In general, surfactants inhibit microbial adhesion to oils and other hydrophobic surfaces. In the present study, we demonstrated that the cationic surfactant cetylpyridinium chloride (CPC) significantly enhanced microbial adhesion to hexadecane and various oils, as well as to the solid hydrophobic surface polystyrene. CPC increased adhesion to hexadecane of Escherichia coli, Candida albicans and Acinetobacter calcoaceticus MR-481 and of expectorated oral bacteria from near 0% to over 90%. The CPC concentration required for optimal enhancement of adhesion was a function of the initial cell density. This phenomenon was inhibited by high salt concentrations and, in the case of E. coli, by a low pH. CPC-pretreated cells were able to bind to hexadecane, but CPC-pretreated hexadecane was unable to bind untreated cells. Another cationic, surface-active antimicrobial agent, chlorhexidine gluconate, was similarly able to promote microbial adhesion to hexadecane. The results suggest that (i) CPC enhances microbial adhesion to hexadecane by binding via electrostatic interactions at the cell surface, thus diminishing surface charge and increasing cell surface hydrophobicity, and (ii) this phenomenon may have applications in oral formulations and in the use of hydrocarbon droplets as a support for cell immobilization. PMID- 16348212 TI - Genetically Engineered Erwinia carotovora: Survival, Intraspecific Competition, and Effects upon Selected Bacterial Genera. AB - Environmental use of genetically engineered microorganisms has raised concerns about potential ecological impact. This research evaluated the survival, competitiveness, and effects upon selected bacterial genera of wild-type and genetically engineered Erwinia carotovora subsp. carotovora to ascertain if differences between the wild-type and genetically engineered strains exist in soil microcosms. The engineered strain contained a chromosomally inserted gene for kanamycin resistance. No significant differences in survival in nonsterile soil over 2 months or in the competitiveness of either strain were observed when the strains were added concurrently to microcosms. For reasons that remain unclear, the engineered strain did survive longer in sterilized soil. The effects of both strains on total bacteria, Pseudomonas and Staphylococcus strains, and actinomycetes were observed. While some apparent differences were observed, they were not statistically significant. A better understanding of the microbial ecology of engineered bacteria, especially pathogens genetically altered for use as biological control agents, is essential before commercial applications can be accomplished. PMID- 16348213 TI - Biodegradation of trichloroethylene in continuous-recycle expanded-bed bioreactors. AB - Experimental bioreactors operated as recirculated closed systems were inoculated with bacterial cultures that utilized methane, propane, and tryptone-yeast extract as aerobic carbon and energy sources and degraded trichloroethylene (TCE). Up to 95% removal of TCE was observed after 5 days of incubation. Uninoculated bioreactors inhibited with 0.5% Formalin and 0.2% sodium azide retained greater than 95% of their TCE after 20 days. Each bioreactor consisted of an expanded-bed column through which the liquid phase was recirculated and a gas recharge column which allowed direct headspace sampling. Pulses of TCE (20 mg/liter) were added to bioreactors, and gas chromatography was used to monitor TCE, propane, methane, and carbon dioxide. Pulsed feeding of methane and propane with air resulted in 1 mol of TCE degraded per 55 mol of substrate utilized. Perturbation studies revealed that pH shifts from 7.2 to 7.5 decreased TCE degradation by 85%. The bioreactors recovered to baseline activities within 1 day after the pH returned to neutrality. PMID- 16348214 TI - Pyrolytic methylation-gas chromatography of whole bacterial cells for rapid profiling of cellular Fatty acids. AB - A novel, on-line derivatization technique has been developed which enables generation of fatty acid methyl ester (FAME) profiles from microorganisms by gas chromatography-mass spectrometry without the need for laborious and time consuming sample preparation. Microgram amounts of bacterial cells are directly applied to a thin ferromagnetic filament and covered with a single drop of methanolic solution of tetramethylammonium hydroxide. After air drying, the filament is inserted into a special gas chromatograph inlet equipped with a high frequency coil, thus enabling rapid inductive heating of the ferromagnetic filament. This so-called Curie-point heating technique is shown to produce patterns of bacterial FAMEs which are qualitatively and quantitatively nearly identical to those obtained from extracts of methylated lipids prepared by conventional sample pretreatment methods. Relatively minor differences involve the loss of hydroxy-substituted fatty acids by the pyrolytic approach as well as strongly enhanced signals of FAMEs derived from mycolic acids. This type of pyrolysis enables on-line derivatization and thermal extraction of volatile derivatives for analysis, whereas the residual components remain on a disposable probe (ferromagnetic wire) of a pyrolytic device. The reduced sample size (micrograms instead of milligrams) and the lack of sample preparation requirements open up the possibility of rapid microbiological identification of single colonies (thus overcoming the need for time-consuming subculturing) as well as analysis of FAME profiles directly from complex environmental samples. PMID- 16348215 TI - Construction and Use of a Nonradioactive DNA Hybridization Probe for Detection of Pseudomonas syringae pv. Tomato on Tomato Plants. AB - Pseudomonas syringae pv. tomato, the causal agent for bacterial speck of tomato, produces the phytotoxin coronatine. A 5.3-kilobase XhoI fragment from the chromosomal region controlling toxin production was cloned into the plasmid pGB2, and the resulting recombinant plasmid, pTPR1, was tested for its ability to serve as a diagnostic probe for P. syringae pv. tomato. In a survey of 75 plant associated bacteria, pTPR1 hybridized exclusively to those strains that produced coronatine. The detection limit for this probe, which was labeled with the Chemiprobe nonradioactive reporter system, was approximately 4 x 10 CFU of lesion bacteria. During the 1989 growing season, a total of 258 leaf and fruit lesions from nine tomato fields were screened for P. syringae pv. tomato by using pTPR1 and the culture method of detection. The best agreement between the two methods, 90%, occurred early in the season with samples taken from relatively young (5 week-old) plants. Young plants also had a higher percentage of P. syringae pv. tomato-positive lesions. P. syringae pv. tomato was the only coronatine producer recovered from the nine tomato fields. All 244 P. syringae pv. tomato strains isolated during this study reacted strongly with the probe. The P. syringae pv. tomato population of healthy field tomato leaves was determined by a pTPR1 colony hybridization procedure. Every probe-positive colony that was isolated and characterized was identified as P. syringae pv. tomato. The pTPR1 probe should expedite disease diagnosis and facilitate epidemiological studies of this pathogen. It also should aid in screening transplant seedlings for bacterial speck infestation. PMID- 16348216 TI - Construction of a simple, inexpensive multiple enzyme-linked immunosorbent assay microdilution plate washer. AB - In this paper, plans are given for the construction of an inexpensive enzyme linked immunosorbent assay plate washer from readily available materials. The wash unit uses an intermittent wash cycle based on a wash manifold cycling over the microdilution plates for a predetermined time. Laboratory tests showed that the unit provided reliable, rapid washing of plates with tap water, with no detectable contamination between wells. Substrate absorbance values for test samples from machine-washed plates were equal to or greater than absorbance values for corresponding samples from plates washed manually by an accepted protocol, by using either enzyme-linked immunosorbent assay wash buffer or tap water. PMID- 16348217 TI - DNA Hybridization Probe for Use in Determining Restricted Nodulation among Bradyrhizobium japonicum Serocluster 123 Field Isolates. AB - Several soybean plant introduction (PI) genotypes have recently been described which restrict nodulation of Bradyrhizobium japonicum serocluster 123 in an apparently serogroup-specific manner. While PI 371607 restricts nodulation of strains in serogroup 123 and some in serogroup 127, those in serogroup 129 are not restricted. When DNA regions within and around the B. japonicum I-110 common nodulation genes were used as probes to genomic DNA from the serogroup strains USDA 123, USDA 127, and USDA 129, several of the probes differentially hybridized to the nodulation-restricted and -unrestricted strains. One of the gene regions, cloned in plasmid pMJS12, was subsequently shown to hybridize to 4.6-kilobase EcoRI fragments from DNAs from nodulation-restricted strains and to larger fragments in nodulation-unrestricted strains. To determine if the different hybridization patterns could be used to predict nodulation restriction, we hybridized pMJS12 to EcoRI-digested genomic DNAs from uncharacterized serocluster 123 field isolates. Of the 36 strains examined, 15 were found to have single, major, 4.6-kilobase hybridizing EcoRI fragments. When tested for nodulation, 80% (12 of 15) of the strains were correctly predicted to be restricted for nodulation of the PI genotypes. In addition, hybridization patterns obtained with pMJS12 and nodulation phenotypes on PI 371607 indicated that there are at least three types of serogroup 127 strains. Our results suggest that the pMJS12 gene probe may be useful in selecting compatible host-strain combinations and in determining the suitability of field sites for the placement of soybean genotypes containing restrictive nodulation alleles. PMID- 16348218 TI - Use of ti plasmid DNA probes for determining tumorigenicity of agrobacterium strains. AB - Probes consisting of T-DNA genes from the Ti plasmid of Agrobacterium tumefaciens were used for determining tumorigenicity of strains. Two P-labeled probes hybridized with 28 of 28 tumorigenic strains of the pathogen but not with 20 of 22 nontumorigenic strains. One probe, pTHE17, consists of all but the far left portion of the T-DNA of strain C58. Probe SmaI7 consists of SmaI fragment 7 of pTiC58, including onc genes 1, 4, and 6a and most of 2. Another probe, pAL4044, consisting of the vir region of strain Ach-5, hybridized with several nontumorigenic as well as tumorigenic strains. Colony hybridizations were done with 28 tumorigenic and 22 nontumorigenic Agrobacterium strains. About 10 CFU of the different tumorigenic strains were detectable with this method. Southern analyses confirmed the presence or absence of Ti plasmids in strains for which tumorigenicity was questioned. Colony hybridization with the T-DNA probes provides a rapid and sensitive means for determining the tumorigenic nature of Agrobacterium strains. PMID- 16348219 TI - Effect of Postemergence, Supplemental Inoculation on Nodulation and Symbiotic Performance of Soybean (Glycine max (L.) Merrill) at Three Levels of Soil Nitrogen. AB - The influence of a supplementary bradyrhizobial inoculation after an initial seed slurry inoculation with the same strain on nodulation and N(2) fixation in soybeans was examined in the greenhouse. The plants were grown in a Typic Eutrocrepts soil: sand mixture containing 25, 65, or 83 mg of N per kg (i.e., native soil N plus N-labeled ammonium sulfate). Harvests were made at early flowering and physiological maturity. The supplementary inoculations which were made 14 or 21 days after planting (DAP) caused formation of substantially more nodules than the single slurry inoculation did. Autoregulation was therefore not completely successful in preventing subsequent infections. For the slurry inoculated plants, at both harvests the proportion of N derived from fixation was greatest in the soil containing the least N, and only slight increases in N(2) fixation resulted from a second inoculation. The inhibition of N(2) fixation at the higher N levels was significantly reduced by a second inoculation at 21 DAP; this treatment resulted in at least a doubling of both the percentage and total amount of N(2) fixed by the single slurry inoculation at physiological maturity. The N(2) fixation increases resulting from the supplementary inoculation at 14 DAP were less pronounced and not significant. Greater N(2) fixation was frequently not reflected by increased total N or dry matter yield, suggesting that the major benefit of the increased fixation was a decreased dependence of plants on soil N for growth. PMID- 16348220 TI - Contributions of Autotrophic and Heterotrophic Nitrifiers to Soil NO and N(2)O Emissions. AB - Soil emission of gaseous N oxides during nitrification of ammonium represents loss of an available plant nutrient and has an important impact on the chemistry of the atmosphere. We used selective inhibitors and a glucose amendment in a factorial design to determine the relative contributions of autotrophic ammonium oxidizers, autotrophic nitrite oxidizers, and heterotrophic nitrifiers to nitric oxide (NO) and nitrous oxide (N(2)O) emissions from aerobically incubated soil following the addition of 160 mg of N as ammonium sulfate kg. Without added C, peak NO emissions of 4 mug of N kg h were increased to 15 mug of N kg h by the addition of sodium chlorate, a nitrite oxidation inhibitor, but were reduced to 0.01 mug of N kg h in the presence of nitrapyrin [2-chloro-6-(trichloromethyl) pyridine], an inhibitor of autotrophic ammonium oxidation. Carbon-amended soils had somewhat higher NO emission rates from these three treatments (6, 18, and 0.1 mug of N kg h after treatment with glucose, sodium chlorate, or nitrapyrin, respectively) until the glucose was exhausted but lower rates during the remainder of the incubation. Nitrous oxide emission levels exhibited trends similar to those observed for NO but were about 20 times lower. Periodic soil chemical analyses showed no increase in the nitrate concentration of soil treated with sodium chlorate until after the period of peak NO and N(2)O emissions; the nitrate concentration of soil treated with nitrapyrin remained unchanged throughout the incubation. These results suggest that chemoautotrophic ammonium oxidizing bacteria are the predominant source of NO and N(2)O produced during nitrification in soil. PMID- 16348221 TI - Mineralization of C-Ring-Labeled Synthetic Lignin Correlates with the Production of Lignin Peroxidase, not of Manganese Peroxidase or Laccase. AB - Recently, Mn(II) has been shown to induce manganese peroxidases (MnPs) and repress lignin peroxidases (LiPs) in defined liquid cultures of several white rot organisms. The present work shows that laccase is also regulated by Mn(II). We therefore used Mn(II) to regulate production of LiP, MnP, and laccase activities while determining the effects of Mn(II) on mineralization of ring-labeled synthetic lignin. At a low Mn(II) level, Phanerochaete chrysosporium and Phlebia brevispora produced relatively high titers of LiPs but only low titers of MnPs. At a high Mn(II) level, MnP titers increased 12- to 20-fold, but LiPs were not detected in crude broths. P. brevispora formed much less LiP than P. chrysosporium, but it also produced laccase activity that increased more than sevenfold at the high Mn(II) level. The rates of synthetic lignin mineralization by these organisms were similar and were almost seven times higher at low than at high Mn(II). Increased synthetic lignin mineralization therefore correlated with increased LiP, not with increased MnP or laccase activities. PMID- 16348222 TI - Detection of horizontal gene transfer by natural transformation in native and introduced species of bacteria in marine and synthetic sediments. AB - Both naturally occurring marine sediments and artificial sediments were used as supports for natural transformation of marine bacteria. While transformation of Pseudomonas stutzeri ZoBell suspended in artificial seawater was not detected when recipient cells and rifampin resistance DNA were loaded onto sterile sediment columns, transformation could be detected at frequencies 4 to 20 times that of spontaneous resistance when recipient cells and rifampin resistance DNA were loaded onto sterile sediment columns. Treatment of these columns with DNase I reduced transformation frequencies to levels comparable to those of spontaneous resistance frequencies. Sediments with higher organic contents supported higher frequencies of transformation than did those with lower amounts of organic matter. Transformation was also detected when recipient cells and DNA were loaded on columns prepared from nonsterile sediments, although the frequencies of transformation were lower than when sterile sediments were used. Finally, nonsterilized sediments that were not supplemented with laboratory strains did not support detectable levels of transformation in sediment columns, but when these same sediments were transferred to filters and placed on complex media, transformation was detected at a frequency three times that for spontaneous resistance. This transformation frequency was partially reduced to levels near that for spontaneous resistance by the addition of DNase I to sediment filters. These results indicate that marine sediments facilitate the uptake and expression of exogenous DNA by transformable marine bacteria and that sediments are a more likely niche for natural transformation than the water column in the marine environment. PMID- 16348223 TI - Anaerobic biodegradation of 2,4,5-trichlorophenoxyacetic Acid in samples from a methanogenic aquifer: stimulation by short-chain organic acids and alcohols. AB - The herbicide 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) was dehalogenated in samples from a methanogenic aquifer to form 2,4- and 2,5-dichlorophenoxyacetic acids as the first detected intermediates. Further incubation of the aquifer slurries resulted in the formation of several intermediates including monochlorophenoxyacetic acids, di- and monochlorophenols, as well as phenol. No transformation of the parent substrate or production of intermediates was detected in autoclaved controls. The pattern of intermediate formation suggested that the anaerobic degradation of 2,4,5-T proceeded by a series of sequential dehalogenation steps with side-chain cleavage reactions occurring at some point before ring cleavage. The addition of short-chain organic acids or alcohols stimulated the onset and rate of 2,4,5-T dehalogenation and decreased the amount of parent substrate still detectable as halogenated intermediates at the end of the experiment. Sulfate addition had the opposite effect on dehalogenation regardless of whether supplemental carbon was added to the aquifer slurries. The inhibitory effect of sulfate on dehalogenation could sometimes be relieved with molybdate, although this effect seemed to be related to the supplemental carbon compound that was used. PMID- 16348224 TI - Purification and Characterization of a Tripeptidase from Lactococcus lactis subsp. cremoris Wg2. AB - A tripeptidase from a cell extract of Lactococcus lactis subsp. cremoris Wg2 has been purified to homogeneity by DEAE-Sephacel and phenyl-Sepharose chromatography followed by gel filtration over a Sephadex G-100 SF column and a high-performance liquid chromatography TSK G3000 SW column. The enzyme appears to be a dimer with a molecular weight of between 103,000 and 105,000 and is composed of two identical subunits each with a molecular weight of about 52,000. The tripeptidase is capable of hydrolyzing only tripeptides. The enzyme activity is optimal at pH 7.5 and at 55 degrees C. EDTA inhibits the activity, and this can be reactivated with Zn, Mn, and partially with Co. The reducing agents dithiothreitol and beta mercaptoethanol and the divalent cation Cu inhibit tripeptidase activity. Kinetic studies indicate that the peptidase hydrolyzes leucyl-leucyl-leucine with a K(m) of 0.15 mM and a V(max) of 151 mumol/min per mg of protein. PMID- 16348225 TI - Differential rates of digestion of bacteria by freshwater and marine phagotrophic protozoa. AB - Differential decreases over time of two bacterial species, Escherichia coli and Enterococcus faecalis, in a freshwater and a marine ecosystem were observed and explained by a differential rate of digestion of these bacteria by phagotrophic flagellates and ciliates. For this purpose, fluorescence-labeled bacteria (FLB) were used and prepared from the two species cited above. The number of FLB was observed for 5 days in fresh and marine waters in the presence or absence (0.2 mum-pore-size-filtered water) of natural microbiota. These experiments showed a longer persistence of Enterococcus faecalis FLB as opposed to Escherichia coli FLB in the presence of natural microbiota. Removal of FLB was due to protozoan grazing because no decrease of FLB number was observed in the absence of natural microbiota. In short-term (about 40 min) ingestion experiments, we found similar clearance rates of Escherichia coli and Enterococcus faecalis FLB by assemblages of flagellates from the freshwater and the marine ecosystem and by cultured assemblages of ciliates from the marine ecosystem. Clearance rates of Enterococcus faecalis FLB were greater than those of Escherichia coli FLB for assemblages of ciliates from the freshwater ecosystem. Comparison of rates of ingestion and digestion of FLB by protozoa showed that Escherichia coli FLB were digested and ingested at similar rates. However, Enterococcus faecalis FLB were digested slower than they were ingested. These results suggest that a longer persistence of Enterococcus faecalis as opposed to Escherichia coli can be explained by a differential digestion by flagellates and ciliates in aquatic ecosystems. Moreover, rates of ingestion and digestion were strongly correlated for both FLB types. PMID- 16348226 TI - Anaerobic Oxidation of Toluene, Phenol, and p-Cresol by the Dissimilatory Iron Reducing Organism, GS-15. AB - The dissimilatory Fe(III) reducer, GS-15, is the first microorganism known to couple the oxidation of aromatic compounds to the reduction of Fe(III) and the first example of a pure culture of any kind known to anaerobically oxidize an aromatic hydrocarbon, toluene. In this study, the metabolism of toluene, phenol, and p-cresol by GS-15 was investigated in more detail. GS-15 grew in an anaerobic medium with toluene as the sole electron donor and Fe(III) oxide as the electron acceptor. Growth coincided with Fe(III) reduction. [ring-C]toluene was oxidized to CO(2), and the stoichiometry of CO(2) production and Fe(III) reduction indicated that GS-15 completely oxidized toluene to carbon dioxide with Fe(III) as the electron acceptor. Magnetite was the primary iron end product during toluene oxidation. Phenol and p-cresol were also completely oxidized to carbon dioxide with Fe(III) as the sole electron acceptor, and GS-15 could obtain energy to support growth by oxidizing either of these compounds as the sole electron donor. p-Hydroxybenzoate was a transitory extracellular intermediate of phenol and p-cresol metabolism but not of toluene metabolism. GS-15 oxidized potential aromatic intermediates in the oxidation of toluene (benzylalcohol and benzaldehyde) and p-cresol (p-hydroxybenzylalcohol and p-hydroxybenzaldehyde). The metabolism described here provides a model for how aromatic hydrocarbons and phenols may be oxidized with the reduction of Fe(III) in contaminated aquifers and petroleum-containing sediments. PMID- 16348227 TI - Rates of microbial metabolism in deep coastal plain aquifers. AB - Rates of microbial metabolism in deep anaerobic aquifers of the Atlantic coastal plain of South Carolina were investigated by both microbiological and geochemical techniques. Rates of [2-C]acetate and [U-C]glucose oxidation as well as geochemical evidence indicated that metabolic rates were faster in the sandy sediments composing the aquifers than in the clayey sediments of the confining layers. In the sandy aquifer sediments, estimates of the rates of CO(2) production (millimoles of CO(2) per liter per year) based on the oxidation of [2 C] acetate were 9.4 x 10 to 2.4 x 10 for the Black Creek aquifer, 1.1 x 10 for the Middendorf aquifer, and <7 x 10 for the Cape Fear aquifer. These estimates were at least 2 orders of magnitude lower than previously published estimates that were based on the accumulation of CO(2) in laboratory incubations of similar deep subsurface sediments. In contrast, geochemical modeling of groundwater chemistry changes along aquifer flowpaths gave rate estimates that ranged from 10 to 10 mmol of CO(2) per liter per year. The age of these sediments (ca. 80 million years) and their organic carbon content suggest that average rates of CO(2) production could have been no more than 10 mmol per liter per year. Thus, laboratory incubations may greatly overestimate the in situ rates of microbial metabolism in deep subsurface environments. This has important implications for the use of laboratory incubations in attempts to estimate biorestoration capacities of deep aquifers. The rate estimates from geochemical modeling indicate that deep aquifers are among the most oligotrophic aquatic environments in which there is ongoing microbial metabolism. PMID- 16348228 TI - Modeling of the bacterial growth curve. AB - Several sigmoidal functions (logistic, Gompertz, Richards, Schnute, and Stannard) were compared to describe a bacterial growth curve. They were compared statistically by using the model of Schnute, which is a comprehensive model, encompassing all other models. The t test and the F test were used. With the t test, confidence intervals for parameters can be calculated and can be used to distinguish between models. In the F test, the lack of fit of the models is compared with the measuring error. Moreover, the models were compared with respect to their ease of use. All sigmoidal functions were modified so that they contained biologically relevant parameters. The models of Richards, Schnute, and Stannard appeared to be basically the same equation. In the cases tested, the modified Gompertz equation was statistically sufficient to describe the growth data of Lactobacillus plantarum and was easy to use. PMID- 16348229 TI - Comparative studies of delignification caused by ganoderma species. AB - Isolates of six species of Ganoderma in the G. lucidum complex were evaluated for their ability to decay wood of Quercus hypoleucoides A. Camus and Abies concolor (Gord. and Glend.) Lindl. ex. Hildebr. by using in vitro agar block decay tests. Morphological, ultrastructural, and chemical studies of decayed wood were used to determine the extent of delignification or simultaneous decay caused by each species of Ganoderma. All species decayed both white fir and oak wood; however, less percent weight loss (%WL) occurred in white fir than oak. In white fir, isolates of two undescribed Ganoderma species (RLG16161, RLG16162, JEA615, and JEA625) caused significantly higher%WL (21 to 26%) than that in G. colossum, G. oregonense, G. meredithiae, and G. zonatum (10 to 16%). Only Ganoderma sp. isolates JEA615 and JEA625 caused delignification, with JEA615 causing a lignin to-glucose gram loss ratio of 1.6:1. Morphological and ultrastructural studies confirmed delignification by this fungus and showed that some delignification had occurred by all of the species, although areas of delignification were limited to small regions adjacent to simultaneously decayed cells. In oak, G. colossum caused significantly less%WL (22 to 35%) than the other species (38 to 52%). All of the species, except G. meredithiae, caused delignification with lignin-to glucose gram loss ratios ranging from 1.4 to 4.9:1. Extensive delignification by isolates of G. colossum and G. oregonense was observed; moderate delignification was caused by the other species. Ganoderma meredithiae caused a simultaneous decay, with only small localized regions of cells delignified, while delignification by G. zonatum was irregular, with specific zones within the cell wall delignified. The thermophilic and chlamydosporic G. colossum has the capacity to cause extensive delignification and appears ideally suited for use in lignin degradation studies and biotechnological applications of lignin-degrading fungi. PMID- 16348230 TI - Mannanase Components from Bacillus pumilus. AB - beta-d-Mannanase from Bacillus pumilus was purified into two components, A and B, which exhibited homogeneity on polyacrylamide gel electrophoresis. These components had molecular weights of 55,000 (A) and 37,000 (B); carbohydrate contents of 15.3% (A) and 7.2% (B); specific activities of 78 (A) and 1,616 (B) U/mg; pH optima of 5.5 to 6.9 (A) and 6.0 (B); and half-lives of 60 (A) and 21 (B) min at 70 degrees C. PMID- 16348231 TI - Several uses for isobutyric Acid-ammonium hydroxide solvent in endotoxin analysis. AB - Many steps in the analysis of rough and semirough endotoxins were found to be facilitated by the use of isobutyric acid-ammonium hydroxide solvent. PMID- 16348232 TI - DNA Is Packaged within Membrane-Derived Vesicles of Gram-Negative but Not Gram Positive Bacteria. AB - Recently, DNA packaged within nuclease-resistant membrane vesicles of Neisseria gonorrhoeae and Borrelia burgdorferi was described. This study assayed 18 species of gram-negative and gram-positive eubacteria for nuclease-protected DNA associated with extracellular membrane vesicles. Vesicles from only the gram negative bacteria contained nuclease-protected linear or supercoiled DNAs or both. PMID- 16348233 TI - DNA Probe for Lactobacillus delbrueckii. AB - From a genomic DNA library of Lactobacillus delbrueckii subsp. bulgaricus, a clone was isolated which complements a leucine auxotrophy of an Escherichia coli strain (GE891). Subsequent analysis of the clone indicated that it could serve as a specific DNA probe. Dot-blot hybridizations with over 40 different Lactobacillus strains showed that this clone specifically recognizes L. delbrueckii subsp. delbrueckii, bulgaricus, and lactis. The sensitivity of the method was tested by using an alpha-P-labeled DNA probe. PMID- 16348234 TI - Purification and Properties of Catechol 1,2-Dioxygenase from Rhizobium leguminosarum biovar viceae USDA 2370. AB - The catechol 1,2-dioxygenase of Rhizobium leguminosarum biovar viceae USDA 2370 was purified 296-fold, yielding a homogeneous preparation with a specific activity of 51.1 U mg of protein. The molecular weight of the native protein was 70,000, with two identical subunits of 34,500 and 1 g-atom of iron per mol. The optimum pH for catalytic activity was 9.0 to 9.5. PMID- 16348235 TI - Characterization of Amylolytic Enzyme Activities Associated with the Hyperthermophilic Archaebacterium Pyrococcus furiosus. AB - The hyperthermophilic archaebacterium Pyrococcus furiosus produces several amylolytic enzymes in response to the presence of complex carbohydrates in the growth medium. These enzyme activities, alpha-glucosidase, pullulanase, and alpha amylase, were detected in both cell extracts and culture supernatants. All activities were characterized by temperature optima of at least 100 degrees C as well as a high degree of thermostability. The existence of this collection of activities in P. furiosus suggests that polysaccharide availability in its growth environment is a significant aspect of the niche from which it was isolated. PMID- 16348236 TI - Interaction of rhizosphere bacteria, fertilizer, and vesicular-arbuscular mycorrhizal fungi with sea oats. AB - Plants must be established quickly on replenished beaches in order to stabilize the sand and begin the dune-building process. The objective of this research was to determine whether inoculation of sea oats (Uniola paniculata L.) with bacteria (indigenous rhizosphere bacteria and N(2) fixers) alone or in combination with vesicular-arbuscular mycorrhizal fungi would enhance plant growth in beach sand. At two fertilizer-N levels, Klebsiella pneumoniae and two Azospirillum spp. did not provide the plants with fixed atmospheric N; however, K. pneumoniae increased root and shoot growth. When a sparingly soluble P source (CaHPO(4)) was added to two sands, K. pneumoniae increased plant growth in sand with a high P content. The phosphorus content of shoots was not affected by bacterial inoculation, indicating that a mechanism other than bacterially enhanced P availability to plants was responsible for the growth increases. When sea oats were inoculated with either K. pneumoniae or Acaligenes denitrificans and a mixed Glomus inoculum, there was no consistent evidence of a synergistic effect on plant growth. Nonetheless, bacterial inoculation increased root colonization by vesicular-arbuscular mycorrhizal fungi when the fungal inoculum consisted of colonized roots but had no effect on colonization when the inoculum consisted of spores alone. K. pneumoniae was found to increase spore germination and hyphal growth of Glomus deserticola compared with the control. The use of bacterial inoculants to enhance establishment of pioneer dune plants warrants further study. PMID- 16348237 TI - Agrobacterium tumefaciens Is a Diazotrophic Bacterium. AB - This is the first report that Agrobacterium tumefaciens can fix nitrogen in a free-living condition as shown by its abilities to grow on nitrogen-free medium, reduce acetylene to ethylene, and incorporate N supplied as N(2). As with most other well-characterized diazotrophic bacteria, the presence of NH(4) in the medium and aerobic conditions repress nitrogen fixation by A. tumefaciens. The system requires molybdenum. No evidence for nodulation was found with pea, peanut, or soybean plants. Further understanding of the nitrogen-fixing ability of this bacterium, which has always been considered a pathogen, should cast new light on the evolution of a pathogenic versus symbiotic relationship. PMID- 16348238 TI - Acid Tolerance of Leuconostoc mesenteroides and Lactobacillus plantarum. AB - In this study, we determined the internal cellular pH response of Leuconostoc mesenteroides and Lactobacillus plantarum to the external pH created by the microorganisms themselves or by lactic or acetic acids and their salts added to the growth medium. Growth of Leuconostoc mesenteroides stopped when its internal pH reached 5.4 to 5.7, and growth of L. plantarum stopped when its internal pH reached 4.6 to 4.8. Variation in growth medium composition or pH did not alter the growth-limiting internal pH reached by these microorganisms. L. plantarum maintained its pH gradient in the presence of either 160 mM sodium acetate or sodium lactate down to an external pH of 3.0 with either acid. In contrast, the DeltapH of Leuconostoc mesenteroides was zero at pH 4.0 with acetate and 5.0 with lactate. No differences were found between d-(-)- and l-(+)-lactic acid for the limiting internal pH for growth of either microorganism. The comparatively low growth-limiting internal pH and ability to maintain a pH gradient at high organic acid concentration may contribute to the ability of L. plantarum to terminate vegetable fermentations. PMID- 16348239 TI - X-Prolyl-Dipeptidyl Aminopeptidase of Lactobacillus delbrueckii subsp. bulgaricus: Characterization of the Enzyme and Isolation of Deficient Mutants. AB - Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397 is able to hydrolyze X proline-para-nitroanilides and X-proline-beta-naphthylamides (X for alanyl- or glycyl-). A single metal-independent cytoplasmic enzyme with a molecular weight estimated to be 82,000 is responsible for these activities and was named X-prolyl dipeptidyl aminopeptidase (X-Pro-DPAP). Isolation and analysis of mutants totally deficient for X-Pro-DPAP activity showed that a total lack of this enzyme induces (i) a decrease in the growth rate; (ii) an increase in cell wall proteinase activity; (iii) the loss of three cell wall proteins with respective molecular masses of 16, 40, and 52 kilodaltons; and (iv) enhancement of a cell wall protein with a molecular mass of 150 kilodaltons. The involvement of X-Pro-DPAP in casein catabolism is discussed. PMID- 16348240 TI - New Field Isolates of Rhizobium leguminosarum Biovar Viciae That Nodulate the Primitive Pea Cultivar Afghanistan in Addition to Modern Cultivars. AB - A collection of 13 field isolates of Rhizobium leguminosarum bv. viciae that have the ability to nodulate the roots of current North American cultivars of peas as well as a "primitive" cultivar, Afghanistan, was examined. These isolates originated in diverse geographical regions of the world, which indicates that this phenotype is not restricted to isolates from any one region. When subclones of the nodulation region from one plasmid were used to examine EcoRI-fragment length polymorphisms in this collection of strains as well as in a collection comprising strains that do not nodulate the primitive cultivar, polymorphism was found in both collections. With one exception, RisphiA6, all strains that nodulated cv. Afghanistan pea contained a region called nodX as an extension to the nodA BCIJ operon that has been observed in all R. leguminosarum bv. viciae strains, including those that do not nodulate cv. Afghanistan pea. RisphiA6 was also the only strain in which nodulating ability could not be associated with a conjugative plasmid. PMID- 16348241 TI - Fate of Ice Nucleation-Active Pseudomonas syringae Strains in Alpine Soils and Waters and in Synthetic Snow Samples. AB - The stability of the ice nucleation activity (INA) and viability of INA Pseudomonas syringae 31a, used as an ice nucleator in the manufacture of synthetic snow, was determined in snow. The viability of P. syringae 1-2b, a rifampin-resistant mutant selected from strain 31a to improve recovery from test samples, was determined in laboratory tests of three alpine soil and water samples from three different sources. Snow samples were exposed to environmental conditions or held in darkness at -20 degrees C. Samples of soil and water were maintained in darkness at 0, 7.5, or 15 degrees C. Parent strain 31a INA decreased significantly (>99.0%) in snow exposed to sunlight and freeze-thaw, while the INA of the cell population in snow held in darkness at -20 degrees C remained essentially unchanged. No viable strain 31a was detected in snow exposed to the environment after 7 days, while the viability of strain 31a in snow held in darkness at -20 degrees C decreased to <3% of the original inoculation at the test conclusion. Mutant strain 1-2b viability was undetectable or had decreased significantly 19 days postinoculation in soil samples held at 0 or 15 degrees C. In contrast, 1-2b viability remained detectable at low levels for the duration of the test in soils held at 7.5 degrees C. The 1-2b population demonstrated a significantly longer half-life in peatlike soil than in the loam soils tested. The rate of decrease in 1-2b viability was essentially the same in the three alpine water samples tested with respect to water temperature and sample location. PMID- 16348242 TI - Rapid quantification of planktonic ciliates: comparison of improved live counting with other methods. AB - THE FOLLOWING EFFICIENT AND QUANTITATIVELY VALID METHOD TO FILTER CONCENTRATE AND COUNT LIVE PLANKTONIC CILIATES WAS DEVELOPED AND COMPARED WITH OTHER TREATMENTS: unconcentrated (raw) samples and centrifuged samples were counted live, and the effects of five different fixatives (HgCl(2), Lugol's iodine, formaldehyde, glutaraldehyde, and Champy-DaFano) on the counts were monitored. Samples originated from a eutrophic mountain lake (Lake Aydat, near Clermont-Ferrand, France). Overall, live filtered counts were similar to counts of raw samples, but they were significantly higher (2 to 2.3 fold, P < 0.05) by analysis of variance than counts from centrifuged samples. Nevertheless, some taxa, i.e., Halteria and Loxodes spp., were sensitive to filtration. The live filtered counts were also comparable to counts of raw HgCl(2)-fixed and settled samples. HgCl(2) and Lugol fixation consistently gave the highest total counts, while significantly lower counts were always obtained with Champy-DaFano-fixed samples. Losses due to fixation were insignificant for raw samples but were substantial and statistically significant in concentrated samples (15% after filtration and 71% after centrifugation, compared with counts from the corresponding live samples). Live counting of passively filter-concentrated ciliates has many advantages over other methods. It is two to four times quicker and more efficient. Ciliates are recognized with certainty, more species are identified, and enumeration of dead organisms (e.g., tintinnid loricas) is avoided. It should be recommended as a quantitatively valid alternative to classical methods for assessing planktonic ciliate populations. PMID- 16348243 TI - Plate assay for simultaneous detection of alginate lyases and determination of substrate specificity. AB - A plate assay to detect the presence of alginate lyases (EC 4.2.2.3) has been developed. The simultaneous use of specific alginate block structures of defined composition allows the substrate specificity of the enzymes to be determined. Clearing zones in the alginate-containing media are visualized with either cetyl pyridinium chloride or ruthenium red. PMID- 16348244 TI - Effect of Methanobrevibacter smithii on Xylanolytic Activity of Anaerobic Ruminal Fungi. AB - Three different ruminal anaerobic fungi, Neocallimastix frontalis PNK2, Sphaeromonas communis B7, and Piromonas communis B19, were grown axenically or in coculture with Methanobrevibacter smithii on xylan. N. frontalis and S. communis in monoculture and coculture accumulated xylobiose, xylose, and arabinose in the growth medium; arabinose was not metabolized, but xylobiose and xylose were subsequently used. The transient accumulation of xylose was much less evident in cocultures. Both the rate and extent of xylan utilization were increased by coculturing, and metabolite profiles became acetogenic as a result of interspecies hydrogen transfer; more acetate and less lactate were formed, while formate and hydrogen did not accumulate. For each of the three fungi, there were marked increases in the specific activities of extracellular xylanase (up to fivefold), alpha-l-arabinofuranosidase (up to fivefold), and beta-d-xylosidase (up to sevenfold) upon coculturing. The stimulating effect on fungal enzymes from coculturing with M. smithii was independent of the growth substrate, and the magnitude of the stimulation varied according to the enzymes and the incubation time. For an N. frontalis-M. smithii coculture, the positive stimulation was maintained during an extended (18-day) incubation period, and this affected not only hemicellulolytic enzymes but also polysaccharidase and glycoside hydrolase enzymes that were not involved in xylan breakdown. The specific activity of cell bound endopeptidase was not increased under the coculture conditions used in this study. The higher enzyme activities in cocultures are discussed in relation to catabolite repression. PMID- 16348245 TI - Spore Yield and Microcycle Conidiation of Colletotrichum gloeosporioides in Liquid Culture. AB - The effect of V8 juice concentration (5 to 40%, vol/vol), spore inoculum density (10 and 10 spores per ml), and liquid batch or fed-batch culture condition on mycelium and spore production by Colletotrichum gloeosporioides was evaluated. The amount of mycelium produced, the time required for initiation of sporulation following attainment of maximum mycelium, and the time for attainment of maximum spore concentration increased with increasing V8 juice concentration in batch culture. Cultures containing V8 juice at >10% achieved a similar spore density (apparent spore-carrying capacity) of about 0.8 mg of spores per ml (1 x 10 to 2 x 10 spores per ml) independent of inoculum density and V8 juice concentration. The relative spore yield decreased from a high of 64% of the total biomass for the low-inoculum 5% V8 culture, through 13% for the analogous 40% V8 culture, to a low of 2% for the high-inoculum 27% V8 culture. Fed-batch cultures were used to establish conditions of high spore density and low substrate availability but high substrate flux. The rate of addition of V8 juice was adjusted to approximate the rate of substrate utilization by the (increasing) biomass. The final spore concentration was about four times higher (3.0 mg of spores per ml) than the apparent spore-carrying capacity in batch culture. This high spore yield was obtained at the expense of greatly reduced mycelium, resulting in a high relative spore yield (62% of the total biomass). Microcycle conidiation occurred in the fed-batch but not batch systems. These data indicate that substrate-limited, fed batch culture can be used to increase the amount and efficiency of spore production by C. gloeosporioides by maintaining microcycle conidiation conditions favoring allocation of nutrients to spore rather than mycelium production. PMID- 16348246 TI - Spectral Irradiance and Distribution of Pigments in a Highly Layered Marine Microbial Mat. AB - The spectral irradiance from 400 to 1,100 nm was measured with depth in the intertidal sand mats at Great Sippewissett Salt Marsh, Mass. These mats contained at least four distinct layers, composed of cyanobacteria, purple sulfur bacteria containing bacteriochlorophyll a (Bchl a), purple sulfur bacteria containing Bchl b, and green sulfur bacteria. Spectral irradiance was measured directly by layering sections of mat on a cosine receptor. Irradiance was also approximated by using a calibrated fiber-optic tip. With the tip, irradiance measurements could be obtained at depth intervals less than 250 mum. The irradiance spectra were correlated qualitatively and quantitatively with the distribution of the diverse chlorophyll pigments in this mat and were compared with spectra recorded in plain sand lacking pigmented phototrophs. We found that the shorter wavelengths (400 to 550 nm) were strongly attenuated in the top 2 mm of the mat. The longer wavelengths (red and near infrared) penetrated to much greater depths, where they were attenuated by Bchl a, b, and c-containing anoxygenic phototrophic bacteria. The specific attenuation bands in the irradiance spectra correlated with the specific in vivo absorption bands of the Bchl-protein complexes in the bacteria. We concluded that the pigments in the phototrophs had a profound affect on the light environment within the mat. It seems likely that the diverse Bchl protein complexes found in the anoxygenic phototrophs evolved in dense mat environments as a result of competition for light. PMID- 16348247 TI - Mutant Strain of Bradyrhizobium japonicum with Increased Symbiotic N(2) Fixation Rates and Altered Mo Metabolism Properties. AB - Mutant strains of Bradyrhizobium japonicum that required higher levels of molybdate than the wild-type strain for growth on NO(3)-containing medium were obtained after transposon Tn5 mutagenesis of the wild-type strain. The mutant strains expressed more than fivefold-greater nitrate reductase activities in the range of 0.1 to 1.0 mM added molybdate compared with activities expressed upon incubation in non-Mo-supplemented medium, whereas the nitrate reductase activity of the wild-type strain (JH) was not markedly influenced by Mo supplementation. In free-living culture, mutant strains JH310 and JH359 expressed substantial nitrogenase activity, even in medium treated to remove molybdate, and nitrogenase activity was influenced little by Mo supplementation, whereas the wild-type strain required 100 nM added Mo for highest nitrogenase activity. Double reciprocal plots of Mo uptake rates versus Mo concentration showed that both bacteroids and free-living cells of mutant strain JH359 had about the same affinity for Mo as did the parent strain. Bacteroids of both the mutants and the wild type also exhibited similar Mo accumulation rates over a 9-min period under very-low-Mo (4 nM) conditions. Nitrogenase activities for strain JH359 and for the wild-type strain in free-living culture were both strongly inhibited by tungsten; thus, the nitrogenase activities of both strains are probably the result of a "conventional" Mo-containing nitrogenase. Soybeans inoculated with strain JH359 and grown under either Mo-supplemented or Mo-deficient conditions had greater specific acetylene reduction rates and significantly greater plant fresh weight than those inoculated with the wild-type strain. Under Mo-deficient conditions, the acetylene reduction rates and plant fresh weights were up to 35 and 58% greater, respectively, for mutant-nodulated plants compared with wild type-strain-nodulated plants. PMID- 16348248 TI - Conjugal Transfer of Megaplasmid 2 between Rhizobium meliloti Strains in Alfalfa Nodules. AB - A DNA fragment containing the RP4 mob function, as well as the gentamicin and spectinomycin resistance genes, was inserted by gene replacement onto the megaplasmid 2 (pM2) of Rhizobium meliloti 0540 (Inf EPS), resulting in PG101 (Inf EPS). The self-transfer of pM2 and the mobilization of pM2 by plasmid RP4-4 were investigated during conjugation between PG101 and R. meliloti 2526 (Nod). In filter conjugations, pM2 was readily mobilized by RP4-4. In addition to this, the self-transfer of one megaplasmid (pM) was detected at a frequency of 3 x 10. Bacteria isolated from the nodules of alfalfa and coinoculated with strains PG101 and 2526 showed that pM2 was mobilized at a frequency of approximately 7 x 10. Bacterial cell numbers were too low in the nodules for detection of the self transfer of pM2 to occur. No pM2 transfer was detected in the inoculum. A comparison of the transfer frequencies for the various conjugation conditions revealed that pM2 transfer occurred as frequently in the nodules as in filter conjugations. These results indicate that the nodule creates conditions for gene transfer that are comparable to optimal laboratory conditions. PMID- 16348249 TI - Dechlorination of Four Commercial Polychlorinated Biphenyl Mixtures (Aroclors) by Anaerobic Microorganisms from Sediments. AB - The rate, extent, and pattern of dechlorination of four Aroclors by inocula prepared from two polychlorinated biphenyl (PCB)-contaminated sediments were compared. The four mixtures used, Aroclors 1242, 1248, 1254, and 1260, average approximately three, four, five, and six chlorines, respectively, per biphenyl molecule. All four Aroclors were dechlorinated with the loss of meta plus para chlorines ranging from 15 to 85%. Microorganisms from an Aroclor 1242 contaminated site in the upper Hudson River dechlorinated Aroclor 1242 to a greater extent than did microorganisms from Aroclor 1260-contaminated sediments from Silver Lake, Mass. The Silver Lake inoculum dechlorinated Aroclor 1260 more rapidly than the Hudson River inoculum did and showed a preferential removal of meta chlorines. For each inoculum the rate and extent of dechlorination tended to decrease as the degree of chlorination of the Aroclor increased, especially for Aroclor 1260. The maximal observed dechlorination rates were 0.3, 0.3, and 0.2 mug-atoms of Cl removed per g of sediment per week for Aroclors 1242, 1248, and 1254, respectively. The maximal observed dechlorination rates for Hudson River and Silver Lake organisms for Aroclor 1260 were 0.04 and 0.21 mug-atoms of Cl removed per g of sediment per week, respectively. The dechlorination patterns obtained suggested that the Hudson River microorganisms were more capable than the Silver Lake organisms of removing the last para chlorine. These results suggest that there are different PCB-dechlorinating microorganisms at different sites, with characteristic specificities for PCB dechlorination. PMID- 16348250 TI - Stereoselective Reduction of Ethyl 4-Chloro-3-Oxobutanoate by a Microbial Aldehyde Reductase in an Organic Solvent-Water Diphasic System. AB - Enzyme-catalyzed asymmetric reduction of ethyl 4-chloro-3-oxobutanoate in an organic solvent-water diphasic system was studied. NADPH-dependent aldehyde reductase isolated from Sporobolomyces salmonicolor AKU4429 and glucose dehydrogenase were used as catalysts for reduction of ethyl 4-chloro-3 oxobutanoate and recycling of NADPH, respectively, in this system. In an aqueous system, the substrate was unstable. Inhibition of the reaction and inactivation of the enzymes by the substrate and the product were also observed. An n-butyl acetate-water diphasic system very efficiently overcame these limitations. In a 1,600-ml-1,600-ml scale diphasic reaction, ethyl (R)-4-chloro-3-hydroxybutanoate (0.80 mol; 86% enantiomeric excess) was produced from the corresponding oxoester in a molar yield of 95.4% with an NADPH turnover of 5,500 mol/mol. PMID- 16348251 TI - Catabolite Inactivation in the Methylotrophic Yeast Pichia pastoris. AB - Inactivation of the alcohol oxidase enzyme system of Pichia pastoris, during the whole-cell bioconversion of ethanol to acetaldehyde, was due to catabolite inactivation. Electron microscopy showed that methanol-grown cells contained peroxisomes but were devoid of these microbodies after the bioconversion. Acetaldehyde in the presence of O(2) was the effector of catabolite inactivation. The process was initiated by the appearance of free acetaldehyde, and was characterized by an increase in the level of cyclic AMP, that coincided with a rapid 55% drop in alcohol oxidase activity. Further enzyme inactivation, believed to be due to proteolytic degradation, then proceeded at a constant but slower rate and was complete 21 h after acetaldehyde appearance. The rate of catabolite inactivation was dependent on acetaldehyde concentration up to 0.14 mM. It was temperature dependent and occurred within 24 h at 37 degrees C and by 6 days at 15 degrees C but not at 3 degrees C. Alcohol oxidase activity was psychrotolerant, with only a 17% decrease in initial specific activity over a temperature drop from 37 to 3 degrees C. In contrast, protease activity was inhibited at temperatures below 15 degrees C. When the bioconversion was run at 3 degrees C, catabolite inactivation was prevented. In the presence of 3 M Tris hydrochloride buffer, 123 g of acetaldehyde per liter was produced at 3 degrees C, compared with 58 g/liter at 30 degrees C. By using 0.5 M Tris in a cyclic batch procedure, 140.6 g of acetaldehyde was produced. PMID- 16348252 TI - Increased Bean (Phaseolus vulgaris L.) Nodulation Competitiveness of Genetically Modified Rhizobium Strains. AB - Rhizobium leguminosarum bv. phaseoli strain collections harbor heterogeneous groups of bacteria in which two main types of strains may be distinguished, differing both in the symbiotic plasmid and in the chromosome. We have analyzed under laboratory conditions the competitive abilities of the different types of Rhizobium strains capable of nodulating Phaseolus vulgaris L. bean. R. leguminosarum bv. phaseoli type I strains (characterized by nif gene reiterations and a narrow host range) are more competitive than type II strains (that have a broad host range), and both types are more competitive than the promiscuous rhizobia isolated from other tropical legumes able to nodulate beans. Type I strains become even more competitive by the transfer of a non-Sym, 225-kilobase plasmid from type II strain CFN299. This plasmid has been previously shown to enhance the nodulation and nitrogen fixation capabilities of Agrobacterium tumefaciens transconjugants carrying the Sym plasmid of strain CFN299. Other type I R. leguminosarum bv. phaseoli transconjugants carrying two symbiotic plasmids (type I and type II) have been constructed. These strains have a diminished competitive ability. The increase of competitiveness obtained in some transconjugants seems to be a transient property. PMID- 16348253 TI - Quantitative microbiological analysis of bacterial community shifts in a high rate anaerobic bioreactor treating sulfite evaporator condensate. AB - The bacterial population of a high-rate, anaerobic, fixed-bed loop reactor treating sulfite evaporator condensate from the pulp industry was studied over a 14-month period. This period was divided into seven cycles that included a startup at the beginning of each cycle. Some 82% of the total biomass was immobilized on and between the porous glass rings filling the reactor. The range of the total number of microorganisms in these biofilms was 2 x 10 to 7 x 10 cells per ml. Enumeration and characterization by microbiological methods and by phase-contrast, epifluorescence, and electron microscopy showed that the samples consisted mainly of the following methanogens: a Methanobacterium sp., a Methanosarcina sp., a Methanobrevibacter sp., and a Methanothrix sp., as well as furfural-degrading sulfate-reducing bacteria resembling Desulfovibrio furfuralis. Viable counts of hydrogenotrophic methanogens were relatively stable (mostly within the range of 3.2 x 10 to 7.5 x 10 cells per ml), but Methanobrevibacter cells increased from <5 to 30% of the total hydrogenotrophic count after transfer of the fixed bed into a second reactor vessel. Acetotrophic methanogens reached their highest numbers of 1.3 x 10 to 2.6 x 10 cells per ml in the last fermentation cycles. They showed a morphological shift from sarcinalike packets in early samples to single coccoid forms in later phases of the fermentation. Furfural-degrading sulfate reducers reached counts of 1 x 10 to 5.8 x 10 cells per ml. The distribution of the chief metabolic groups between free fluid and biofilms was analyzed in the fifth fermentation cycle: 4.5 times more furfural degraders were found in the free fluid than in the biofilms. In contrast, 5.8 times more acetotrophic and 16.6 times more hydrogenotrophic methanogens were found in the biofilms than in the free liquid. The data concerning time shifts of morphotypes among the trophic groups of methanogens corroborated the trends observed by using immunological assays on the same samples. PMID- 16348254 TI - Enhanced Nodulation and Nitrogen Fixation by a Revertant of a Nodulation Defective Bradyrhizobium japonicum Tryptophan Auxotroph. AB - In greenhouse studies, the symbiotic properties of a prototrophic revertant (TA11 NOD) of a nodulation defective tryptophan auxotroph of Bradyrhizobium japonicum were compared with those of the normally nodulating wild-type strain, B. japonicum I-110 ARS. Strain I-110 ARS was the parent of auxotrophic mutant TA11. Plants inoculated with TA11 NOD contained significantly more nitrogen per plant than did plants inoculated with wild-type bacteria (275.9 +/- 35 versus 184 +/- 18 mg). Also, plants that received the revertant were larger, averaging 8.4 +/- 0.9 g (dry weight) versus 6.4 +/- 0.6 g for those that received the wild-type bacterial strain. Additionally, plants that received the NOD strain had 56% more nodules and 41% more nodule mass than did control plants. With both inocula, average nodule size and amount of nitrogen fixed per gram of nodule were about the same. These data indicated that the improvement in nitrogen fixation observed with the TA11 NOD resulted from an increase in the overall nodule number. The physiological basis for this increase in nodulation is not known, but enhanced tryptophan catabolism does not appear to be involved. PMID- 16348255 TI - Transfer of Isolated Nuclei into Protoplasts of Trichoderma harzianum. AB - Protoplasts released from young hyphae of Trichoderma harzianum contained 0 to 10 nuclei per protoplast, and most (about 80%) contained from 4 to 6 nuclei. Most protoplasts were larger than 3 mum in diameter. Nuclei were isolated from protoplasts of an auxotrophic mutant of T. harzianum and transferred into protoplasts obtained from another auxotroph of the same strain. This intrastrain nuclear transfer gave rise to numerous progeny which were stable, prototrophic, and heterokaryotic. Interstrain transfers in which nuclei from a wild-type prototroph of one strain were transferred into protoplasts from a lysine deficient auxotroph of a second strain were also done. Heterokaryotic progeny were recovered from these interstrain transfers when the regenerating protoplasts were provided with a low concentration of lysine 48 h after the initial plating. Heterokaryotic progeny contained 11 to 17% of donor-type nuclei. Progeny homokaryotic for donor-type nuclei were obtained as single-spore isolates. These homokaryotic isolates expressed the isozyme pattern and colony morphology phenotype of the nuclear donor. When regenerating protoplasts were provided with lysine 10 days after the initial plating, only a single progeny was obtained. However, single-spore subprogeny of this nuclear transfer were prototrophic and exhibited a wide range of unstable morphological phenotypes. PMID- 16348256 TI - Regulation of Sugar Transport Systems in Fusarium oxysporum var. lini. AB - Fusarium oxysporum var. lini (ATCC 10960) formed a facilitated diffusion system for glucose (K(s), about 10 mM) when grown under repressed conditions. Under conditions of derepression, the same system was present together with a high affinity (K(s), about 40 muM) active system. The maximum velocity of the latter was about 5% of that of the facilitated diffusion system. The high-affinity system was under the control of glucose repression and glucose inactivation. When lactose was the only carbon source in the medium, a facilitated diffusion system for lactose was found (K(s), about 30 mM). PMID- 16348257 TI - Description of a Novel Symbiotic Bacterium from the Brittle Star, Amphipholis squamata. AB - A gram-negative, marine, facultatively anaerobic bacterial isolate designated strain AS-1 was isolated from the subcuticular space of the brittle star, Amphipholis squamata. Its sensitivity to O/129 and novobiocin, overall morphology, and biochemical characteristics and the moles percent guanine-plus cytosine composition of its DNA (42.9 to 44.4) suggest that this isolate should be placed in the genus Vibrio. Strain AS-1 was not isolated from ambient seawater and is distinct from described Vibrio species. This symbiotic bacterium may assist its host as one of several mechanisms of nutrient acquisition during the brooding of developing embryos. PMID- 16348258 TI - Characterization of Root Surface and Endorhizosphere Pseudomonads in Relation to Their Colonization of Roots. AB - An extensive colonization of the endorhizosphere by fluorescent pseudomonads was observed in tomato plants grown on artificial substrates. These studies reveal that a significantly higher percentage of pseudomonads obtained from the endorhizosphere (30%) reduced plant growth than those obtained from the root surface (4%). Lipopolysaccharide patterns, cell envelope protein patterns, and other biochemical characteristics indicated that Pseudomonas isolates obtained from the endorhizosphere are distinct from Pseudomonas isolates obtained from the root surface. Isolates from the endorhizosphere especially were able to recolonize the endorhizosphere of both sterile and nonsterile tomato roots. The ability of the endorhizosphere isolates to colonize the endorhizosphere significantly correlated with their agglutination by tomato root agglutinin but did not correlate with chemotaxis to seed exudates of tomato. No correlation between colonization of the endorhizosphere and agglutination by root agglutinin could be demonstrated for the root surface isolates. We propose that agglutination of specific Pseudomonas strains by root agglutinin is of importance in the initial phase of adherence of bacteria to the root surface. PMID- 16348259 TI - Transport of beta-Galactosides in Lactobacillus plantarum NC2. AB - The ability of Lactobacillus plantarum NC2 to transport thiomethyl-beta-d galactoside in the presence or absence of various inhibitors was investigated to determine the mechanism of beta-galactoside transport in this bacterium. A novel system employing l-arabinose as an energy-generating compound is described, and evidence that this transport is energized by an ATP-driven proton motive force is presented. PMID- 16348260 TI - Toxicity of 1,1,1-Trichloroethane and Trichloroethene on a Mixed Culture of Methane-Oxidizing Bacteria. AB - The influence of trichloroethene (TCE; 0 to 65 mg/liter) and 1,1,1 trichloroethane (1,1,1-TCA; 0 to 103 mg/liter) on methane consumption of a mixed culture of methane-oxidizing bacteria was studied in laboratory batch experiments. Increasing concentrations of TCE or 1,1,1-TCA resulted in decreasing methane consumption. Methane consumption was totally inhibited at a concentration of 13 mg of TCE per liter, while methane consumption was still observed at the upper studied concentration of 103 mg of 1,1,1-TCA per liter. The inhibition of methane consumption by TCE depended on the initial concentration of methane. A model accounting for competitive inhibition between methane and TCE or 1,1,1-TCA was used to simulate methane consumption at various concentrations of TCE or 1,1,1-TCA. The simulations indicated that competitive inhibition may be the mechanism causing the inhibitory effect of TCE on methane consumption, while this does not seem to be the case for 1,1,1-TCA. PMID- 16348261 TI - Effects of Culture Conditions on Poly(beta-Hydroxybutyric Acid) Production by Haloferax mediterranei. AB - The halobacterium Haloferax mediterranei accumulates poly(beta-hydroxybutyrate) (PHB) as intracellular granules. The conditions for PHB production in batch and continuous cultures have been studied and optimized. Phosphate limitation is essential for PHB accumulation in large quantities. Glucose and starch are the best carbon sources. With 2% starch, 0.00375% KH(2)PO(4), and 0.2% NH(4)Cl in batch culture, a production of ca. 6 g of PHB per liter was reached, being 60% of the total biomass dry weight, and giving a yield over the carbon source of 0.33 g/g. The PHB production in continuous cultures was stable over a 3-month period. Our results demonstrate that H. mediterranei is an interesting candidate for industrial production of biological polyesters. PMID- 16348262 TI - Interrelationship of Xylanase Induction and Cellulase Induction of Trichoderma longibrachiatum. AB - Xylose oligomers rapidly induced xylanase activity of Trichoderma longibrachiatum, whereas induction was delayed in the presence of glucose. Cellobiose, cellopentaose, and xylobiose did not induce detectable levels of cellulase activity. However, mixtures of xylobiose with cellobiose or cellopentaose rapidly induced cellulase activity. In addition, mixtures of xylobiose with cellopentaose or cellobiose induced xylanase activity more effectively than xylobiose alone. Both xylanase and cellulase activity were detected after a lag period in the presence of lactose. PMID- 16348263 TI - Effectiveness of Different Frankia Cell Types as Inocula for the Actinorhizal Plant Casuarina. AB - The soil bacterium Frankia of the Actinomycetales, capable of forming N(2)-fixing symbiotic root nodules on a diverse array of actinorhizal plants, has several morphological forms when grown in pure culture. Fresh hydrated preparations of whole cells, hyphae, and spores were all infective on seedlings of Casuarina at different dilutions. Desiccated hyphae showed no infection capacity, while desiccated spores remained infective, although at a reduced level. On the basis of most-probable-number statistics, spore suspensions were 3 orders of magnitude more infective than hyphae. PMID- 16348264 TI - Biodegradation of Halogenated Hydrocarbon Fumigants by Nitrifying Bacteria. AB - Three species of nitrifying bacteria were tested for the ability to degrade the halocarbon fumigants methyl bromide, 1,2-dichloropropane, and 1,2-dibromo-3 chloropropane. The soil nitrifiers Nitrosomonas europaea and Nitrosolobus multiformis degraded all three fumigants, while the marine nitrifier Nitrosococcus oceanus degraded only methyl bromide under the conditions tested. Inhibition of biodegradation by allylthiourea and acetylene, specific inhibitors of ammonia monooxygenase, suggests that ammonia monooxygenase is the enzyme which catalyzes fumigant degradation. PMID- 16348265 TI - Characterization of the Opine-Utilizing Microflora Associated with Samples of Soil and Plants. AB - Microorganisms utilizing an opine as the sole carbon source were recovered from crown gall tumors, soil, and surface-disinfected potato tubers. The effect of the opines octopine, nopaline, succinamopine, and mannopine as selective substrates was compared with that of the auxin indoleacetic acid. Selection on octopine and indoleacetic acid favored the fluorescent pseudomonads, whereas mannopine allowed the frequent recovery of agrobacteria. Coryneforms which utilized succinamopine or mannopine were detected in soil, but not in tumors. Fungi growing on succinamopine or mannopine and a mannopine-utilizing Pseudomonas putida were isolated from tumor and soil, respectively. PMID- 16348266 TI - Bacterial Accumulation of Metallic Compounds. AB - Xanthomonas maltophilia accumulated metals or metallic compounds within the cell in the form of one or a few particles, in a peculiar condition similar to that of an electrolyte solution in an electric cell. The particles consisted of many little granules, most of which were assumed to be composed mainly of a compound of zinc and phosphorus in an amorphous state. PMID- 16348267 TI - Occurrence of an Affinity Site apart from the Active Site on the Raw-Starch Digesting but Non-Raw-Starch-Adsorbable Bacillus subtilis 65 alpha-Amylase. AB - alpha-Cyclodextrin specifically inhibited raw starch digestion by Bacillus subtilis 65 alpha-amylase. The raw starch digestibility and alpha-cyclodextrin Sepharose 6B adsorbability of this alpha-amylase were simultaneously lost when the specific domain corresponding to the affinity site essential for raw starch digestion was deleted by proteolysis. Occurrence of the affinity site on raw starch-digesting enzymes was proven also with bacterial amylase. PMID- 16348268 TI - Carbon Metabolism Enzymes of Rhizobium meliloti Cultures and Bacteroids and Their Distribution within Alfalfa Nodules. AB - Several carbon metabolism enzymes were measured in cultured cells and bacteroids of Rhizobium meliloti 102F51 and in alfalfa root nodule cytosol. The enzyme activity levels of the pentose phosphate pathway were much higher than those of the Embden-Meyerhof-Parnas or Entner-Doudoroff pathways in extracts of cultured cells. The pattern of enzyme activities in the bacteroids was different from that of cultured cells. PMID- 16348269 TI - Nitrogen Fixation Dynamics of Two Diazotrophic Communities in Mono Lake, California. AB - [This corrects the article on p. 614 in vol. 56.]. PMID- 16348270 TI - Effects of organic substrates on dechlorination of aroclor 1242 in anaerobic sediments. AB - The effects of different organic substrates on the abilities of anaerobic sediment enrichments to reductively dechlorinate polychlorinated biphenyls (PCBs) were studied. Sediments collected from a site previously contaminated with PCBs were dosed with additional PCBs (Aroclor 1242; approximately 300 ppm [300 mug/g], sediment dry weight) and incubated anaerobically with acetate, acetone, methanol, or glucose. The pattern of dechlorination was similar for each substrate-fed batch; however, the extents and rates of dechlorination were different. Significant dechlorination over time was observed, with the relative rates and extents of dechlorination being greatest for methanol-, glucose-, and acetone-fed batches and least for acetate-fed batches. Dechlorination occurred primarily on the meta- and para- positions of the highly chlorinated congeners, resulting in the accumulation of less-chlorinated, primarily ortho-substituted products. No significant dechlorination was observed in incubation batches receiving no additional organic substrate, even though identical inorganic nutrients were added to all incubation batches. In addition, dechlorination was not observed in autoclaved controls that received substrate and nutrients. PMID- 16348271 TI - Interactions between Bacillus thuringiensis subsp. israelensis and Fathead Minnows, Pimephales promelas Rafinesque, under Laboratory Conditions. AB - Interactions between Bacillus thuringiensis subsp. israelensis and fathead minnows, Pimephales promelas, were studied in laboratory exposures to two commercial formulations, Vectobac-G and Mosquito Attack. Mortality among fatheads exposed to 2.0 x 10 to 6.5 x 10 CFU/ml with both formulations was attributed to severe dissolved oxygen depletion due to formulation ingredients rather than to direct toxicity from the parasporal crystal. No adverse effects were observed at 6.4 x 10 CFU/ml and below. Fathead minnows rapidly accumulated high numbers of spores with 1 h of exposure to 2.2 x 10 CFU of Mosquito Attack per ml, producing whole-body counts of 4.0 x 10 CFU per fish. Comparison of counts on gastrointestinal tract samples and whole-body samples and high numbers of spores in feces indicated that ingestion was the major route of exposure. B. thuringiensis subsp. israelensis spore counts decreased rapidly after transfer of fish to clean water, with a drop of over 3 orders of magnitude in 1 day. Spores were rarely detected in fish after 8 days but were detectable in feces for over 2 weeks. These findings suggest that fish could influence the dissemination of B. thuringiensis subsp. israelensis, and possibly other microbial agents, in the aquatic environment. PMID- 16348272 TI - Formation and Action of Lignin-Modifying Enzymes in Cultures of Phlebia radiata Supplemented with Veratric Acid. AB - Transformation of veratric (3,4-dimethoxybenzoic) acid by the white rot fungus Phlebia radiata was studied to elucidate the role of ligninolytic, reductive, and demeth(ox)ylating enzymes. Under both air and a 100% O(2) atmosphere, with nitrogen limitation and glucose as a carbon source, reducing activity resulted in the accumulation of veratryl alcohol in the medium. When the fungus was cultivated under air, veratric acid caused a rapid increase in laccase (benzenediol:oxygen oxidoreductase; EC 1.10.3.2) production, which indicated that veratric acid was first demethylated, thus providing phenolic compounds for laccase. After a rapid decline in laccase activity, elevated lignin peroxidase (ligninase) activity and manganese-dependent peroxidase production were detected simultaneously with extracellular release of methanol. This indicated apparent demethoxylation. When the fungus was cultivated under a continuous 100% O(2) flow and in the presence of veratric acid, laccase production was markedly repressed, whereas production of lignin peroxidase and degradation of veratryl compounds were clearly enhanced. In all cultures, the increases in lignin peroxidase titers were directly related to veratryl alcohol accumulation. Evolution of CO(2) from 3 OCH(3)-and 4-OCH(3)-labeled veratric acids showed that the position of the methoxyl substituent in the aromatic ring only slightly affected demeth(ox)ylation activity. In both cases, more than 60% of the total C was converted to CO(2) under air in 4 weeks, and oxygen flux increased the degradation rate of the C-labeled veratric acids just as it did with unlabeled cultures. PMID- 16348273 TI - Effect of sulfate and organic carbon supplements on reductive dehalogenation of chloroanilines in anaerobic aquifer slurries. AB - When di-, tri-, and tetrachloroaniline were incubated in methanogenic groundwater slurries, they were reductively dehalogenated by the aquifer microbiota. 2,3,4 Trichloroaniline was metabolized by two pathways. Primary dehalogenation occurred at either the meta or ortho position of this substrate to form 2,4- and 3,4 dichloroaniline, respectively. The latter chemical could be stoichiometrically converted to 3-chloroaniline. 2,3,4,5-Tetrachloroaniline was degraded by the sequential removal of halogens from the para and then the ortho position to form 3,5-dichloroaniline. An additional pathway was observed with this substrate when the aquifer slurries were amended with butyrate. That is, halogens could be removed from both the meta and ortho positions of tetrachloroaniline. The amendment of sulfate to methanogenic aquifer slurries slowed the rate of 2,3,4,5 tetrachloroaniline degradation and increased the amount of substrate channeled through the additional pathway. The reported intermediates or end products are identified by their chromatographic mobility and mass-spectral profiles. PMID- 16348274 TI - Acetoin Fermentation by Citrate-Positive Lactococcus lactis subsp. lactis 3022 Grown Aerobically in the Presence of Hemin or Cu. AB - CitrLactococcus lactis subsp. lactis 3022 produced more biomass and converted most of the glucose substrate to diacetyl and acetoin when grown aerobically with hemin and Cu. The activity of diacetyl synthase was greatly stimulated by the addition of hemin or Cu, and the activity of NAD-dependent diacetyl reductase was very high. Hemin did not affect the activities of NADH oxidase and lactate dehydrogenase. These results indicated that the pyruvate formed via glycolysis would be rapidly converted to diacetyl and that the diacetyl would then be converted to acetoin by the NAD-dependent diacetyl reductase to reoxidize NADH when the cells were grown aerobically with hemin or Cu. On the other hand, the Y(Glu) value for the hemincontaining culture was lower than for the culture without hemin, because acetate production was repressed when an excess of glucose was present. However, in the presence of lipoic acid, an essential cofactor of the dihydrolipoamide acetyltransferase part of the pyruvate dehydrogenase complex, hemin or Cu enhanced acetate production and then repressed diacetyl and acetoin production. The activity of diacetyl synthase was lowered by the addition of lipoic acid. These results indicate that hemin or Cu stimulates acetyl coenzyme A (acetyl-CoA) formation from pyruvate and that lipoic acid inhibits the condensation of acetyl-CoA with hydroxyethylthiamine PP(i). In addition, it appears that acetyl-CoA not used for diacetyl synthesis is converted to acetate. PMID- 16348275 TI - Modification of water-soluble coal-derived products by dibenzothiophene-degrading microorganisms. AB - To study mechanisms by which microorganisms oxidize thiophenic sulfur in coal, we tested bacterial cultures for the ability to degrade dibenzothiophene (DBT), DBT 5-oxide, and DBT-sulfone and to modify water-soluble coal products derived from Illinois no. 6 and Ugljevik coals. In yeast extract medium, the majority of selected isolates degraded DBT and accumulated DBT-5-oxide in culture fluids; all but one of the cultures degraded DBT-5-oxide, and none of them degraded DBT sulfone. Elemental analysis data indicated that the microbial cultures were able to decrease the amount of sulfur in soluble coal products derived from Illinois no. 6 and Ugljevik coals. However, these data suggested that microbially mediated sulfur removal from soluble Ugljevik coal occurred by nonspecific mechanisms. That is, extensive degradation of the carbon structure was concurrent with the loss of sulfur. This conclusion was supported by X-ray photoelectron spectroscopic data which indicated that the reduced sulfur forms in the soluble Ugljevik coal product was not oxidized by microbial treatment. PMID- 16348276 TI - Effect of Agitation on Ligninase Activity and Ligninase Production by Phanerochaete chrysosporium. AB - The white rot fungus Phanerochaete chrysosporium produces extracellular ligninases as part of its idiophasic ligninolytic system. Agitation has been widely reported to suppress both ligninase production and lignin degradation. Results show that mechanical inactivation of ligninase is possibly the reason why ligninase accumulation is low or absent in agitated shake-flask cultures. Agitation seems to affect the catalytic activity of ligninase and has no apparent effect on either the rate of ligninase production or the physiology of P. chrysosporium. The detergents Tween 20, Tween 40, Tween 60, Tween 80, and 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) are able to protect both purified ligninase and extant ligninase in culture fluids (free of biomass) against mechanical inactivation due to agitation. Addition of Tween 80 at the end of primary growth to agitated shake flasks containing either pelleted or immobilized mycelial cultures results in production and maintenance of high levels of ligninase activity over several days under conditions of high agitation. Possible mechanisms by which the detergents could protect ligninase are discussed. PMID- 16348277 TI - Coexisting bacterial populations responsible for multiphasic mineralization kinetics in soil. AB - Experiments were conducted to study populations of indigenous microorganisms capable of mineralizing 2,4-dinitrophenol (DNP) in two soils. Previous kinetic analyses indicated the presence of two coexisting populations of DNP-mineralizing microorganisms in a forest soil (soil 1). Studies in which eucaryotic and procaryotic inhibitors were added to this soil indicated that both populations were bacterial. Most-probable-number counts with media containing different concentrations of DNP indicated that more bacteria could mineralize low concentrations of DNP than could metabolize high concentrations of it. Enrichments with varying concentrations of DNP and various combinations of inhibitors consistently resulted in the isolation of the same two species of bacteria from soil 1. This soil contained a large number and variety of fungi, but no fungi capable of mineralizing DNP were isolated. The two bacterial isolates were identified as a Janthinobacterium sp. and a Rhodococcus sp. The Janthinobacterium sp. had a low mu(max) and a low K(m) for DNP mineralization, whereas the Rhodococcus sp. had much higher values for both parameters. These differences between the two species of bacteria were similar to differences seen when soil was incubated with different concentrations of DNP. Values for mu(max) from soil incubations were similar to mu(max) values obtained in pure culture studies. In contrast, K(s) and K(m) values showed greater variation between soil and pure culture studies. The results of this study help to confirm predictions that two physiologically distinct bacterial populations are responsible for the multiphasic mineralization kinetics observed in the soil studied. PMID- 16348278 TI - Adhesion of cellulolytic ruminal bacteria to barley straw. AB - Adhesion of the cellulolytic ruminal bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes to barley straw was measured by incubating bacterial suspensions with hammer-milled straw for 30 min, filtering the mixtures through sintered glass filters, and measuring the optical densities of the filtrates. Maximum adhesion of both species occurred at pH 6.0 and during mid- to late exponential phase. Adhesion was saturable at 33 and 23 mg (dry weight) g of straw for R. flavefaciens and F. succinogenes, respectively. Methyl cellulose and carboxymethyl cellulose inhibited adhesion by 24 to 33%. Competition between species was determined by measuring characteristic cell-associated enzyme activities in filtrates of mixtures incubated with straw; p-nitrophenyl-beta-d lactopyranoside hydrolysis was used as a marker for F. succinogenes, while either beta-xylosidase or carboxymethyl cellulase was used for R. flavefaciens, depending on the other species present. R. flavefaciens had no influence on F. succinogenes adhesion, and F. succinogenes had only a minor (<20%) effect on R. flavefaciens adhesion. The noncellulolytic ruminal bacteria Bacteroides ruminicola and Selenomonas ruminantium had no influence on adhesion of either cellulolytic species, although these organisms also adhered to the straw. We concluded that R. flavefaciens and F. succinogenes have separate, specific adhesion sites on barley straw that are not obscured by competition with non cellulolytic species. PMID- 16348279 TI - Risk Assessment Studies: Detailed Host Range Testing of Wild-Type Cabbage Moth, Mamestra brassicae (Lepidoptera: Noctuidae), Nuclear Polyhedrosis Virus. AB - The host range of a multiply enveloped nuclear polyhedrosis virus (NPV) (Baculoviridae) isolated from the cabbage moth, Mamestra brassicae (Lepidoptera: Noctuidae), was determined by challenging a wide range of insect species with high (10 polyhedral inclusion bodies) and low (10 polyhedral inclusion bodies) doses of the virus. The identity of the progeny virus was confirmed by dot blotting. Analysis of 50% lethal dose was carried out on selected species, and the progeny virus was identified by using restriction enzyme analysis and Southern blotting. Other than the Lepidoptera, none of the species tested was susceptible to M. brassicae NPV. Within the Lepidoptera, M. brassicae NPV was infective to members of four families (Noctuidae, Geometridae, Yponomeutidae, and Nymphalidae). Of 66 lepidopterous species tested, M. brassicae NPV was cross infective to 32 of them; however, 91% of the susceptible species were in the Noctuidae. The relevance of host range data in risk assessment studies is discussed. PMID- 16348280 TI - Formation of stable bdelloplasts as a starvation-survival strategy of marine bdellovibrios. AB - Several wild-type isolates of marine bdellovibrios formed stable bdelloplasts when they infected gram-negative bacterial prey under certain culture conditions. Synchronous predator-prey cultures and low nutrient concentrations increased the yield of stable bdelloplasts. The bdellovibrio cells retained in the stable bdelloplasts showed a high survival capacity in nutrient-depleted saline solution (10% viable Bdellovibrio cells after 3 months at 25 degrees C), whereas Bdellovibrio attack-phase cells kept under the same starvation conditions lost viability more quickly (1% viable cells after 48 h). The addition of yeast extract to a stable bdelloplast suspension induced lysis of the bdelloplasts and release of motile infecting attack-phase Bdellovibrio cells. Other substances, such as free amino acids, protein hydrolysates, NH(4), carbohydrates, and organic amines, did not induce such a release. Stable bdelloplasts were highly hydrophobic and had a lower endogenous respiration rate than attack-phase cells. In general, stable bdelloplasts were almost as sensitive to temperature changes, desiccation, sonication, tannic acid, and Triton X-100 treatment as attack-phase cells. Electron microscopy of stable bdelloplasts did not reveal any extra cell wall layer, either in the bdelloplast envelope or in the retained Bdellovibrio cells, unlike the bdellocysts of the soil bacterium Bdellovibrio sp. strain W. We propose that formation of stable bdelloplasts is a survival strategy of marine bdellovibrios which occurs in response to nutrient- and prey-poor seawater habitats. PMID- 16348281 TI - Stability of Integrated Plasmids in the Chromosome of Lactococcus lactis. AB - Derivatives of plasmids pBR322, pUB110, pSC101, and pTB19, all containing an identical fragment of lactococcal chromosomal DNA, were integrated via a Campbell like mechanism into the same chromosomal site of Lactococcus lactis MG1363, and the transformants were analyzed for the stability of the integrated plasmids. In all cases the erythromycin resistance gene of pE194 was used as a selectable marker. Transformants obtained by integration of the pBR322 derivatives contained a head-to-tail arrangement of several plasmid copies, which most likely was caused by integration of plasmid multimers. Single-copy integrations were obtained with the pSC101 and pTB19 derivatives. In all of these transformants no loss of the erythromycin gene was detected during growth for 100 generations in the absence of the antibiotic. In contrast, transformants containing integrated amplified plasmid copies of pUB110 derivatives were unstable under these conditions. Since pUB110 appeared to have replicative activity in L. lactis, we suggest that this activity destabilized the amplified structures in L. lactis. PMID- 16348282 TI - Hemoproteins of Bradyrhizobium japonicum Cultured Cells and Bacteroids. AB - The hemoprotein content of 17 strains of Bradyrhizobium japonicum bacteroids from field-grown plants and the corresponding strains of cultured cells was determined spectrally. The major terminal oxidases, cytochromes (cyt) aa(3) and o, were present in all strains of cultured cells. cyt aa(3) was present in significant amounts in bacteroids only in strains of DNA homology group II. cyt o appeared to be present in bacteroids of all strains, and the average level was the same as in cultured cells. cyt b and c in the membrane fractions were higher in bacteroids of all strains compared with cultured cells. cyt P-450 was present in both the membrane and soluble fractions of bacteroids of most strains. The total P-450 content varied sixfold among strains. A CO-reactive hemoprotein, P-422, was present in the soluble fraction of all strains of cultured cells. P-422 may be a hemoglobinlike protein, and it was present in significant amounts in bacteroids only in DNA homology group I strains. PMID- 16348283 TI - Enzyme Activities Affecting End Product Distribution by Lactobacillus plantarum in Response to Changes in pH and O(2). AB - Lactobacillus plantarum catabolic end products changed in response to environmental conditions. While lactate was always the major end product, acetate was produced in alkaline and aerobic environments. Acetoin levels decreased under alkaline conditions. Changes in acetoin dehydrogenase, acetate kinase, NADH oxidase, pyruvate oxidase, and acetate kinase activities correlated with changes in end product distribution. PMID- 16348284 TI - Insecticidal activity of a bacterial crystal protein expressed by a recombinant baculovirus in insect cells. AB - Baculoviruses are insect pathogens with a relatively slow speed of action, and this has limited their use as control agents of insect pests. Introduction into baculoviruses of genes which code for proteins interfering specifically with insect metabolism or metamorphosis, such as toxins, hormones, and enzymes, may enhance the pathogenicity of these viruses. The complete insecticidal crystal protein gene cryIA(b) of Bacillus thuringiensis subsp. aizawai 7.21 was engineered into the nuclear polyhedrosis virus of Autographa californica (AcNPV) in place of the polyhedrin gene. In infected Spodoptera frugiperda cells, the cryIA(b) gene was expressed at a high level without interference with AcNPV production. The crystal protein was found in the cytoplasm of S. frugiperda cells, mainly as large crystals with an ultrastructure similar to that of B. thuringiensis crystals. Infected-cell extracts inhibited feeding of the large cabbage white Pieris brassicae. The toxicity of the crystal protein expressed by AcNPV recombinants was comparable with that of the crystal protein expressed by a corresponding Escherichia coli recombinant. PMID- 16348285 TI - Enhanced Propionate Formation by Propionibacterium freudenreichii subsp. freudenreichii in a Three-Electrode Amperometric Culture System. AB - In order to influence the fermentation pattern of Propionibacterium freudenreichii towards enhanced propionate formation, growth and product formation with glucose and lactate as energy sources were studied in a three electrode poised-potential amperometric culture system. With anthraquinone 2,6 disulfonic acid (E(0)' = -184 mV; poised electron potential = -224 mV) or cobalt sepulchrate (E(0)' = -350 mV; -390 mV) as mediator and an activated platinum working electrode, reduction of bacterially oxidized mediator occurred fast enough to keep more than 50% of the respective mediator (in minimum 0.4 mM) in the reduced state, up to a current of 2 mA. With glucose as substrate, 90.0 or 97.3% propionate was formed during exponential growth in the presence of 0.5 mM anthraquinone 2,6-disulfonic acid or 0.4 mM cobalt sepulchrate, respectively. Growth yields of 56.3 or 53.8 g of cell material per mol of substrate degraded were calculated, respectively, and the electrons were transferred quantitatively from the working electrode to the bacterial cells. With l-lactate, only 68.6 or 72.9% propionate was formed with the same mediators. The results are discussed with respect to energetics, electron transfer potentials, and potential application of the new technique in technical propionate production. PMID- 16348286 TI - Binding of Colloidal MnO(2) by Extracellular Polysaccharides of Pedomicrobium manganicum. AB - The extracellular acidic polysaccharides of the manganese-oxidizing bacterium Pedomicrobium manganicum were able to bind preformed colloidal MnO(2). The capacity of the cells to bind MnO(2) was pH dependent. Enhanced binding capacity below pH 5 suggests that ionic bonding forces are involved in the binding mechanism and that there is a charge reversal on the acidic polysaccharides between pH 5 and 4 that is due to increased protonation of carboxyl groups. PMID- 16348287 TI - Characterization of Egg Yolk Antibodies for Detection and Quantification of Selenomonas ruminantium by Using an Enzyme-Linked Immunosorbent Assay. AB - The specificity of polyclonal antibodies prepared against strains of Selenomonas ruminantium, the effect of assay conditions, and quantification of individual strains in mixed-cell suspensions of selenomonad strains were examined in this study. Whole-cell suspensions were prepared with pure cultures of S. ruminantium PC18, HD(4), GA192, and D. Each cell suspension was injected into a Leghorn laying hen, and polyclonal antibodies were harvested from eggs laid in week 3 or 7 following initial immunization. Antibodies made to the S. ruminantium strains readily discerned the homologous strain from the heterologous strains. Cross reactivity among antibodies and the heterologous S. ruminantium strains ranged from 5 to 26%. Among non-S. ruminantium species, cross-reactivity of S. ruminantium antibodies was greatest with Selenomonas sputigena (3 to 34%) and Succinivibrio dextrinosolvens (0 to 37%). Antibodies made to strains GA192 and D were used to quantify a mixture of the two strains. Both antibodies responded to graded concentrations of the homologous antigen in the biculture mixtures in accord with the change in the direct cell counts for each strain (strain D, R = 0.92; strain GA192, R = 0.90). This enzyme-linked immunosorbent assay enabled concurrent and accurate quantification of two strains of S. ruminantium subsp. ruminantium in a mixed-cell suspension with a precision of much less than 1 order of magnitude. PMID- 16348288 TI - Growth and Maintenance of Thiobacillus ferrooxidans Cells. AB - A rapid and sensitive spectrophotometric procedure was developed for monitoring the growth of Thiobacillus ferrooxidans in liquid culture. Values determined for the optical densities at 500 nm of washed T. ferrooxidans cell suspensions were directly proportional to both total cell number and total cell protein concentration and provided an accurate measurement of culture growth rate. The utility of this procedure was demonstrated by conducting physiological studies on the influence of CO(2) and FeSO(4) availability on the growth of T. ferrooxidans. In addition, we describe a procedure for the long-term maintenance of cells T. ferrooxidans that ensures culture purity and genetic stability. PMID- 16348289 TI - Regulation of Dissimilatory Fe(III) Reduction Activity in Shewanella putrefaciens. AB - Under anaerobic conditions, Shewanella putrefaciens is capable of respiratory chain-linked, high-rate dissimilatory iron reduction via both a constitutive and inducible Fe(III)-reducing system. In the presence of low levels of dissolved oxygen, however, iron reduction by this microorganism is extremely slow. Fe(II) trapping experiments in which Fe(III) and O(2) were presented simultaneously to batch cultures of S. putrefaciens indicated that autoxidation of Fe(II) was not responsible for the absence of Fe(III) reduction. Inhibition of cytochrome oxidase with CN resulted in a high rate of Fe(III) reduction in the presence of dissolved O(2), which suggested that respiratory control mechanisms did not involve inhibition of Fe(III) reductase activities or Fe(III) transport by molecular oxygen. Decreasing the intracellular ATP concentrations by using an uncoupler, 2,4-dinitrophenol, did not increase Fe(III) reduction, indicating that the reduction rate was not controlled by the energy status of the cell. Control of electron transport at branch points could account for the observed pattern of respiration in the presence of the competing electron acceptors Fe(III) and O(2). PMID- 16348290 TI - Transformation of Rhodococcus fascians by High-Voltage Electroporation and Development of R. fascians Cloning Vectors. AB - The analysis of the virulence determinants of phytopathogenic Rhodococcus fascians has been hampered by the lack of a system for introducing exogenous DNA. We investigated the possibility of genetic transformation of R. fascians by high voltage electroporation of intact bacterial cells in the presence of plasmid DNA. Electrotransformation in R. fascians D188 resulted in transformation frequencies ranging from 10/mug of DNA to 10/mug of DNA, depending on the DNA concentration. The effects of different electrical parameters and composition of electroporation medium on transformation efficiency are presented. By this transformation method, a cloning vector (pRF28) for R. fascians based on an indigenous 160-kilobase (chloramphenicol and cadmium resistance-encoding) plasmid pRF2 from strain NCPPB 1675 was developed. The origin of replication and the chloramphenicol resistance gene on pRF28 were used to construct cloning vectors that are capable of replication in R. fascians and Escherichia coli. The electroporation method presented was efficient enough to allow detection of the rare integration of replication-deficient pRF28 derivatives in the R. fascians D188 genome via either homologous or illegitimate recombination. PMID- 16348291 TI - Potential of Agrobacterium tumefaciens and Octopine-Utilizing Fluorescent Pseudomonas Strains To Attach to Susceptible Potato Tissues. AB - The binding characteristics of two octopine-catabolizing pseudomonads, Pseudomonas fluorescens B99A and E175D, which were isolated from crown galls, have been examined. The binding of strain B99A to potato disks was very weak, followed a Freundlich isotherm, and was temperature and pH independent. Strain E175D displayed strong attachment and followed a Langmuir isotherm. Despite these fundamental differences in binding characteristics, when each strain was placed in competitive binding assays with either Agrobacterium tumefaciens B6 or A. tumefaciens ATCC 15955, the number of bound pseudomonad cells decreased compared with those obtained in independent trials. Furthermore, the binding of A. tumefaciens cells was increased. In prebinding experiments, in which the potato disks were bound with the pseudomonads before exposure to the agrobacteria, the number of bound pseudomonad cells again decreased. This implies that increased desorption was occurring. In these prebinding studies, the numbers of bound A. tumefaciens ATCC 15955 increased, but the number of bound A. tumefaciens B6 remained the same. The mechanism for this observed synergism on the binding of agrobacterial cells and the depression in bound pseudomonad cells is believed to be alterations in the electrostatic or ionic charges on the plant and bacterial cell surfaces. The synergistic effect on A. tumefaciens undermines the use of these pseudomonads as potential biocontrol agents for crown gall. PMID- 16348292 TI - Growth of Octopine-Catabolizing Pseudomonas spp. under Octopine Limitation in Chemostats and Their Potential To Compete with Agrobacterium tumefaciens. AB - The growth characteristics of five octopine-catabolizing pseudomonads have been determined in batch and continuous cultures. All five strains belonged to rRNA homology group I and showed a more psychrotrophic growth pattern than did Agrobacterium tumefaciens B6 and ATCC 15955. In chemostats limited by octopine, either as the source of carbon and nitrogen or the sole source of nitrogen, maximum specific growth rates and substrate affinities were lower than those in chemostats limited by glutamate. These growth dynamics were similar to those observed for Agrobacterium strains B6 and ATCC 15955 even though the catabolic genes and pathways are believed to be different in the two genera. An analysis of the yields in octopine-limited chemostats indicated that the use of octopine as the sole source of carbon and nitrogen was grossly inefficient. Octopine and presumably lysopine and octopinic acid provided a better source of nitrogen than of carbon. One of the Pseudomonas fluorescens strains, E175D, was able to produce its highest yield on octopine as a nitrogen source. Competition models formulated on pure culture parameters indicated that two of the Pseudomonas spp. would dominate A. tumefaciens B6 and ATCC 15955 when in simple competition for octopine as a limiting substrate. PMID- 16348293 TI - Competition of Octopine-Catabolizing Pseudomonas spp. and Octopine-Type Agrobacterium tumefaciens for Octopine in Chemostats. AB - The ability of two octopine-catabolizing Pseudomonas spp. and two virulent octopine-type Agrobacterium tumefaciens to compete for substrates has been examined in chemostats. In dual cultures with octopine or glutamate as the limiting carbon or nitrogen source, Pseudomonas fluorescens B99A and E175D always dominated over A. tumefaciens B6 or ATCC 15955. The growth dynamics of each strain in pure culture indicated that some form of antagonism was occurring in dual culture to permit the predominance of the pseudomonads under certain conditions. Although both pseudomonads fluoresce, pyoverdine was not responsible for the observed inhibition. An unidentified antibiotic secreted by both pseudomonads is believed to be responsible. A. tumefaciens B6 grew synergistically in the presence of P. fluorescens B99A with octopine as the limiting nitrogen source. This behavior of Agrobacterium strain B6 may help overcome its grossly inefficient use of octopine as previously reported. The ability of these two pseudomonads to outcompete the agrobacteria under all conditions tested raises the possibility that under field conditions, infectious agrobacteria may be succeeded by opine-catabolizing pseudomonads around crown gall tumors and in the rhizosphere. PMID- 16348294 TI - Limiting an Insect Infestation of Nitrogen-Fixing Root Nodules of the Pigeon Pea (Cajanus cajan) by Engineering the Expression of an Entomocidal Gene in Its Root Nodules. AB - A region of DNA which determined the production of the insecticidal toxin of Bacillus thuringiensis subsp. israelensis was cloned into a derivative of a broad host-range group IncQ plasmid vector of gram-negative bacteria. The plasmid which we constructed was transferred by conjugative mobilization into a Bradyrhizobium species that nodulates pigeon peas. In this species the construction was maintained stably in the absence of selection and expressed the gene that was installed. Experiments in a greenhouse with the strain which we constructed indicated that this organism provides protection against root nodule damage by the larvae of the insect Rivellia angulata (Diptera). PMID- 16348295 TI - Preferential osmolyte accumulation: a mechanism of osmotic stress adaptation in diazotrophic bacteria. AB - A common cellular mechanism of osmotic-stress adaptation is the intracellular accumulation of organic solutes (osmolytes). We investigated the mechanism of osmotic adaptation in the diazotrophic bacteria Azotobacter chroococcum, Azospirillum brasilense, and Klebsiella pneumoniae, which are adversely affected by high osmotic strength (i.e., soil salinity and/or drought). We used natural abundance C nuclear magnetic resonance spectroscopy to identify all the osmolytes accumulating in these strains during osmotic stress generated by 0.5 M NaCl. Evidence is presented for the accumulation of trehalose and glutamate in Azotobacter chroococcum ZSM4, proline and glutamate in Azospirillum brasilense SHS6, and trehalose and proline in K. pneumoniae. Glycine betaine was accumulated in all strains grown in culture media containing yeast extract as the sole nitrogen source. Alternative nitrogen sources (e.g., NH(4)Cl or casamino acids) in the culture medium did not result in measurable glycine betaine accumulation. We suggest that the mechanism of osmotic adaptation in these organisms entails the accumulation of osmolytes in hyperosmotically stressed cells resulting from either enhanced uptake from the medium (of glycine betaine, proline, and glutamate) or increased net biosynthesis (of trehalose, proline, and glutamate) or both. The preferred osmolyte in Azotobacter chroococcum ZSM4 shifted from glutamate to trehalose as a consequence of a prolonged osmotic stress. Also, the dominant osmolyte in Azospirillum brasilense SHS6 shifted from glutamate to proline accumulation as the osmotic strength of the medium increased. PMID- 16348296 TI - Direct Comparison of Phosphate Uptake by Adnate and Loosely Attached Microalgae within an Intact Biofilm Matrix. AB - We report a direct comparison of phosphate uptake by adnate and loosely attached microalgae in an intact biofilm matrix, with resolution at the level of individual cells. Track scanning electron microscope autoradiography enabled assay of [P]phosphate uptake from the overlying water by adnate algae left undisturbed on mature leaves of the macrophyte Potamogeton illinoensis or on artificial plant mimics. The epiphyte communities developed in either phosphate poor or moderately phosphate-enriched water, and they were assayed on both natural and artificial plants. All adnate taxa examined from both natural and artificial plants in both habitats took up significantly less radiolabel when assayed beneath the overlying matrix than when they were exposed to the water upon removal of the overstory material. Track scanning electron microscope autoradiography and track light microscope autoradiography were intercalibrated to enable comparison of [P]phosphate uptake by adnate and loosely attached components of the epiphyte matrix. Loosely attached cells on substrata from both habitats took up significantly more radiolabel than did underlying adnate cells, indicating that access to phosphate supplies from the water depended on the position of microbial cells in the matrix. In this short-term assay, the adnate microalgae were relatively isolated from the water column nutrient source. PMID- 16348297 TI - Floating filters, a novel technique for isolation and enumeration of fastidious, acidophilic, iron-oxidizing, autotrophic bacteria. AB - Nuclepore polycarbonate filters floating on a liquid, FeSO(4)-containing medium (pH 1.6) were used to isolate a moderately thermophilic bacterium from a pyrite oxidizing enrichment culture. The isolate failed to grow on any of the conventional solid media tried. To test the general applicability of the method, the enumeration of a fastidious acidophilic organism, Thiobacillus ferrooxidans, was carried out and the results compared with those obtained with other filters, solid media, and the most probable number technique. T. ferrooxidans showed better viability on the floating polycarbonate filters and grew in a much shorter time (4 to 5 days) than with the other techniques. PMID- 16348298 TI - Characterization of thermoanaerobacter glucose isomerase in relation to saccharidase synthesis and development of single-step processes for sweetener production. AB - Regulation of glucose isomerase synthesis was studied in Thermoanaerobacter strain B6A, which fermented a wide variety of carbohydrates including glucose, xylose, lactose, starch, and xylan. Glucogenic amylase activities and beta galactosidase were produced constitutively, whereas the synthesis of glucose isomerase was induced by either xylose or xylan. Production of these saccharidase activities was not significantly repressed by the presence of glucose or 2 deoxyglucose in the growth media. Glucose isomerase production was optimized by controlling the culture pH at 5.5 during xylose fermentation. The apparent temperature and pH optima for these cell-bound saccharidase activities were as follows: glucose isomerase, 80 degrees C, pH 7.0 to 7.5; glucogenic amylase, 70 degrees C, pH 5.0 to 5.5; and beta-galactosidase, 60 degrees C, pH 6.0 to 6.5 Glucose isomerase, glucogenic amylase, and beta-galactosidase were produced in xylose-grown cells that were active and stable at 60 to 70 degrees C and pH 6.0 to 6.5. Under single-step process conditions, these saccharidase activities in whole cells or cell extracts converted starch or lactose directly into fructose mixtures. A total of 96% of initial liquefied starch was converted into a 49:51 mixture of glucose and fructose, whereas 85% of initial lactose was converted into a 40:31:29 mixture of galactose, glucose, and fructose. PMID- 16348299 TI - Distribution and rate of methane oxidation in sediments of the Florida everglades. AB - Rates of methane emission from intact cores were measured during anoxic dark and oxic light and dark incubations. Rates of methane oxidation were calculated on the basis of oxic incubations by using the anoxic emissions as an estimate of the maximum potential flux. This technique indicated that methane oxidation consumed up to 91% of the maximum potential flux in peat sediments but that oxidation was negligible in marl sediments. Oxygen microprofiles determined for intact cores were comparable to profiles measured in situ. Thus, the laboratory incubations appeared to provide a reasonable approximation of in situ activities. This was further supported by the agreement between measured methane fluxes and fluxes predicted on the basis of methane profiles determined by in situ sampling of pore water. Methane emissions from peat sediments, oxygen concentrations and penetration depths, and methane concentration profiles were all sensitive to light-dark shifts as determined by a combination of field and laboratory analyses. Methane emissions were lower and oxygen concentrations and penetration depths were higher under illuminated than under dark conditions; the profiles of methane concentration changed in correspondence to the changes in oxygen profiles, but the estimated flux of methane into the oxic zone changed negligibly. Sediment-free, root-associated methane oxidation showed a pattern similar to that for methane oxidation in the core analyses: no oxidation was detected for roots growing in marl sediment, even for roots of Cladium jamaicense, which had the highest activity for samples from peat sediments. The magnitude of the root-associated oxidation rates indicated that belowground plant surfaces may not markedly increase the total capacity for methane consumption. However, the data collectively support the notion that the distribution and activity of methane oxidation have a major impact on the magnitude of atmospheric fluxes from the Everglades. PMID- 16348300 TI - Heat Resistance Correlated with DNA Content in Bacillus megaterium Spores. AB - Two subpopulations of Bacillus megaterium spores (1.360 and 1.355 g/ml) were obtained by density gradient centrifugation. The heavier spores had a higher thermoresistance (e.g., D(80) = 186 versus 81 min) and a higher DNA content (1.25 x 10 versus 0.65 x 10 g per spore, apparently corresponding to digenomic versus monogenomic spores). No appreciable differences were found in the mineral and dipicolinic acid contents or in the inactivation kinetics of the two subpopulations. The implications of the findings are discussed with regard to mechanisms of heat resistance and of inactivation. PMID- 16348301 TI - Isolation of roquefortine C from feed grain. AB - Roquefortine C was isolated from feed grain heavily infected by Penicillium roqueforti. The identity of the mycotoxin was confirmed by mass spectrometry. Other mycotoxins that are known to be produced by P. roqueforti such as PR toxin, patulin, and penicillic acid were not detected in the grain. PMID- 16348302 TI - Isolation and enological characterization of malolactic bacteria from the vineyards of northwestern Spain. AB - Thirty-five strains of malolactic bacteria were isolated from grapes and alcoholic and malolactic fermentations in two vineyards from northwestern Spain. These belonged to six species of the genera Lactobacillus and Leuconostoc. The results of their partial enological characterization showed that 47.5% utilized more than 80% of the initial malic acid. PMID- 16348303 TI - Method designed to detect alginate-degrading bacteria. AB - A simple turbidimetric method was developed to detect alginate degradation. Bacteria were grown in alginate-containing media, and culture fluids were mixed with an acidic albumin solution. Failure to develop a white turbidity indicated an alginate degrader. The method showed alginate degradation by Vibrio alginolyticus ATCC 17749, in contrast to prior descriptions. PMID- 16348304 TI - Selection of Astaxanthin-Overproducing Mutants of Phaffia rhodozyma with beta Ionone. AB - beta-Ionone, an end ring analog of beta-carotene, inhibits astaxanthin production in the red yeast Phaffia rhodozyma. Astaxanthin-overproducing mutants of this yeast are easily spotted on beta-ionone-containing yeast malt agar plates. beta Ionone appears to block astaxanthin synthesis at the beta-carotene level. PMID- 16348305 TI - Isolation and Structural Identification of a New Metabolite of Fusarium equiseti. AB - A fluorescent compound was isolated and purified from rice cultures of Fusarium equiseti (Alaska 2-2). Mass spectrometry and nuclear magnetic resonance data indicated that its structure is 2,2-dimethyl-5-amino-6-(3'-hydroxyl-4'-methoxyl butyryl)-4-chromone. It is an analog of the mycotoxin fusarochromanone, in which the amino group on C-3' is replaced by a hydroxyl group and the hydroxyl group on C-4' is replaced by a methoxyl group. PMID- 16348306 TI - Rapid detection of chlorine-induced bacterial injury by the direct viable count method using image analysis. AB - [This corrects the article on p. 389 in vol. 56.]. PMID- 16348307 TI - Nucleotide Sequence and Distribution of the pTR2030 Resistance Determinant (hsp) Which Aborts Bacteriophage Infection in Lactococci. AB - [This corrects the article on p. 2257 in vol. 56.]. PMID- 16348308 TI - Anaerobic Aryl Reductive Dehalogenation of Halobenzoates by Cell Extracts of "Desulfomonile tiedjei". AB - We studied the transformation of halogenated benzoates by cell extracts of a dehalogenating anaerobe, "Desulfomonile tiedjei." We found that cell extracts possessed aryl reductive dehalogenation activity. The activity was heat labile and dependent on the addition of reduced methyl viologen, but not on that of reduced NAD, NADP, flavin mononucleotide, flavin adenine dinucleotide, desulfoviridin, cytochrome c(3), or benzyl viologen. Dehalogenation activity in extracts was stimulated by formate, CO, or H(2), but not by pyruvate plus coenzyme A or by dithionite. The pH and temperature optima for aryl dehalogenation were 8.2 and 35 degrees C, respectively. The rate of dehalogenation was proportional to the amount of protein in the assay mixture. The substrate specificity of aryl dehalogenation activity for various aromatic compounds in "D. tiedjei" cell extracts was identical to that of whole cells, except differences were observed in the relative rates of halobenzoate transformation. Dehalogenation was 10-fold greater in "D. tiedjei" extracts prepared from cells cultured in the presence of 3-chlorobenzoate, suggesting that the activity was inducible. Aryl reductive dehalogenation in extracts was inhibited by sulfite, sulfide, and thiosulfate, but not sulfate. Experiments with combinations of substrates suggested that cell extracts dehalogenated 3 iodobenzoate more readily than either 3,5-dichlorobenzoate or 3-chlorobenzoate. Dehalogenation activity was found to be membrane associated. This is the first report characterizing aryl dehalogenation activity in cell extracts of an obligate anaerobe. PMID- 16348309 TI - Lignocarbohydrate solubilization from straw by actinomycetes. AB - Actinomycetes grown on wheat straw solubilized a lignocarbohydrate fraction which could be recovered by acid precipitation. Further characterization of this product (APPL) during growth of Streptomyces sp. strain EC1 revealed an increase in carboxylic acid and phenolic hydroxyl content, suggesting progressive modification. This was also observed in dioxane-extracted lignin fractions of degraded straw, and some similarity was further suggested by comparative infrared spectroscopy. However, the molecular weight profile of APPL was relatively constant during growth of Streptomyces sp. strain EC1 on straw, while analysis of the dioxane-extracted lignin fractions appeared to show fragmentation followed by repolymerization. Lignocarbohydrate solubilization could be monitored in all cultures by routine assay of APPL-associated protein, which accounted for up to 20% of the extracellular culture protein in some cases. Interestingly, this protein fraction was found to include active hydrolytic and oxidative enzymes involved in the degradation of lignocellulose, and specific enzyme activities were often increased in the acid-insoluble fractions of culture supernatants. This was particularly important for peroxidase and veratryl oxidase activities, which could be readily detected in the acid-precipitable lignocarbohydrate complex but were virtually undetectable in untreated culture supernatants. PMID- 16348310 TI - Biomass and Biological Activity during the Production of Compost Used as a Substrate in Mushroom Cultivation. AB - The production of a suitable substrate for the cultivation of the common white button mushroom, Agaricus bisporus, is referred to as composting. High microbiological activity causes temperatures of the composting material to rise as high as 80 degrees C. At stacking, an optimal oxygen consumption rate of 140 mumol of O(2) h g (dry weight) was found in the compost at 50 degrees C, whereas the oxygen consumption rate of the end product was lower at all temperatures tested. No significant differences were observed between biomass content and mineralization rate of C-labeled glutamate of the two composts. Biomass content was shown to be a major function of both temperature and the sampling site position in the stack. On the basis of the results reported here, a minimal composting time of 3.3 days for the phase I process was calculated. Further suggestions are made to reduce the time necessary for the production of a substrate for A. bisporus considerably. PMID- 16348311 TI - Competitive Ability and Efficiency in Nodule Formation of Strains of Bradyrhizobium japonicum. AB - In the American Midwest, superior N(2)-fixing inoculant strains of Bradyrhizobium japonicum consistently fail to produce the majority of nodules on the roots of field-grown soybean. Poor nodulation by inoculant strains is partly due to their inability to stay abreast of the expanding soybean root system in numbers sufficient for them to be competitive with indigenous bradyrhizobia. However, certain strains are noncompetitive even when numerical dominance is not a factor. In this study, we tested the hypothesis that the nodule occupancy achieved by strains is related to their nodule-forming efficiency. The nodulation characteristics and competitiveness of nine strains of B. japonicum were compared at both 20 and 30 degrees C. The root tip marking technique was used, with the nodule-forming efficiency of each strain estimated from the average position of the uppermost nodule and the number of nodules formed above the root tip mark. The competitiveness of the nine strains relative to B. japonicum USDA 110 was determined by using immunofluorescence to identify nodule occupants. The strains differed significantly in competitiveness with USDA 110 and in nodulation characteristics, strains that were poor competitors usually proving to be inferior in both the average position of the uppermost root nodule and the number of nodules formed above the root tip mark. Thus, competitiveness was correlated with both the average position of the uppermost nodule (r = 0.5; P = 0.036) and the number of nodules formed above the root tip mark (r = 0.64; P = 0.005), while the position of the uppermost nodule was also correlated to the percentage of plants nodulated above the root tip mark (r = 0.81; P < 0.001) and the percentage of plants nodulated on the taproot (r = 0.67; P = 0.002). PMID- 16348312 TI - Genetic Transformation of the Biocontrol Fungus Gliocladium virens to Benomyl Resistance. AB - Methodology was developed to isolate and regenerate protoplasts from the biocontrol fungus Gliocladium virens and to transform them to benomyl resistance with a Neurospora crassa beta-tubulin gene. Southern blots demonstrated that multiple copies of the vector integrated into the chromosomal DNA of stable biotypes but not of abortive transformants. Analysis of nuclear condition in vegetative and asexual structures demonstrated that no structure of G. virens is dependably uninucleate and thus preferentially suitable for transformation. PMID- 16348313 TI - Co-occlusion and persistence of a baculovirus mutant lacking the polyhedrin gene. AB - A co-occlusion process was evaluated as a commercially and ecologically acceptable strategy for the development of genetically improved baculovirus insecticides. Coinfection of Spodoptera frugiperda (IPLB-SF-21) tissue culture cells with Autographa californica nuclear polyhedrosis virus (AcMNPV) and an AcMNPV mutant (Ac-E10) lacking the polyhedrin gene resulted in occlusion of both virus types within polyhedra. The amount of occluded Ac-E10 virions in progeny polyhedra populations during serial passage in Trichoplusia ni larvae was evaluated. Maintenance of the mutant in progeny polyhedra required polyhedra inocula containing equal numbers of the two virus types at a high dose. A significant reduction in occluded mutant nucleocapsids occurs with inoculum levels below a 100% lethal dose. At inoculum levels below a 30% lethal dose, the majority of fourth-instar larvae were infected with only one type of virus. The commercial application and ecological advantages of the co-occlusion process are discussed. PMID- 16348314 TI - Mathematical Model of Thermal Destruction of Bacillus stearothermophilus Spores. AB - The experimental survival curves of Bacillus stearothermophilus spores in aqueous suspension, for six constant temperatures ranging from 105 to 130 degrees C, displayed an initial shoulder before a linear decline. To interpret these observations, we supposed that, before the heat treatment, the designated spore suspension contained a countable and mortal N(0) population of activated spores and an M(0) population of dormant spores which remained masked during spore counting and had to be activated before being destroyed by heat. We also hypothesized that the mechanisms of both activation and destruction are, at constant temperature, ruled by first-order kinetics, with velocity constants k(A) and k(D), respectively. Mathematical analysis showed that this model could represent not only our experimental survival curves, but also all other shapes (linear and biphasic) of survival curves found in the literature; also, there is an inherent symmetry in the model formulation between the activation and destruction reactions, and we showed that the dormancy rate (tau = M(0)/N(0)) is the only parameter which permits a distinction between the two reactions. By applying the model to our experimental data and considering that the dormancy rate is not dependent on the treatment temperature, we showed that, for the studied suspension, the limiting reaction was the activation reaction. PMID- 16348315 TI - Effects of Physicochemical Factors on the Adhesion to Cellulose Avicel of the Ruminal Bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes subsp. succinogenes. AB - Ruminococcus flavefaciens adhered instantly to cellulose, while Fibrobacter succinogenes had the highest percentage of adherent cells after about 25 min of contact between bacteria and cellulose. Adhesion of R. flavefaciens was unaffected by high concentrations of sugars (5%), temperature, pH, oxygen, metabolic inhibitors, and lack of Na. In contrast, the attachment was affected by the removal of divalent cations (Mg and Ca), the presence of cellulose derivatives (methylcellulose and hydroxyethylcellulose), and cystine. Adhesion of F. succinogenes was sensitive to low and high temperatures, high concentrations of glucose and cellobiose (5%), hydroxyethylcellulose (0.1%), redox potential, pH, lack of monovalent cations, and the presence of an inhibitor of membrane ATPases or lasalocid and monensin. Cells of F. succinogenes heated at 100 degrees C no longer were adherent. On the other hand, adhesion was insensitive to the lack of divalent cations (Mg and Ca), the presence of 2,4-dinitrophenol, tetrachlorosalicylanilide, or inhibitors of the electron transfer chains. Adhesion of F. succinogenes seems to be related to the metabolic functions of the cell. External proteins and/or cellulases themselves might play a part in the attachment process. Several mechanisms are probably involved in the adhesion of R. flavefaciens, the main one being the interaction between the large glycocalyx and the divalent cations Ca and Mg. Hydrophobic bonds and enzymes may also be involved. PMID- 16348316 TI - Selective Medium for Isolating Phanerochaete chrysosporium from Soil. AB - A selective medium was developed that is capable of isolating Phanerochaete chrysosporium from soil. This medium contains 15 ppm of benomyl (15 mug g) and 550 ppm of streptomycin sulfate in 2% malt agar and is held at 39 degrees C after inoculation. P. chrysosporium was isolated from three nonsterile forest soils to which the fungus had been added. These soils contained large microbial populations. PMID- 16348317 TI - In Situ Depletion of Pentachlorophenol from Contaminated Soil by Phanerochaete spp. AB - The ability of two white rot fungi to deplete pentachlorophenol (PCP) from soil, which was contaminated with a commercial wood preservative, was examined in a field study. Inoculation of soil containing 250 to 400 mug of PCP g with either Phanerochaete chrysosporium or P. sordida resulted in an overall decrease of 88 to 91% of PCP in the soil in 6.5 weeks. This decrease was achieved under suboptimal temperatures for the growth and activity of these fungi, and without the addition of inorganic nutrients. Since the soil had a very low organic matter content, peat was included as a source of organic carbon for fungal growth and activity. A small percentage (8 to 13%) of the decrease in the amount of PCP was a result of fungal methylation to pentachloroanisole. Gas chromatographic analysis of sample extracts did not reveal the presence of extractable transformation products other than pentachloroanisole. Thus, when losses of PCP via mineralization and volatilization were negligible, as they were in laboratory scale studies (R. T. Lamar, J. A. Glaser, and T. K. Kirk, Soil Biol. Biochem. 22:433-440, 1990), most of the PCP was converted to nonextractable soil-bound products. The nature, stability, and toxicity of soil-bound transformation products, under a variety of conditions, must be elucidated before use of these fungi in soil remediation efforts can be considered a viable treatment method. PMID- 16348318 TI - Pulsed-Field Gel Electrophoresis of SmaI Digests of Lactococcal Genomic DNA, a Novel Method of Strain Identification. AB - The pulsed-field gel electrophoresis (PFGE) pattern of SmaI digests of 29 strains of Lactococcus lactis subsp. lactis and subsp. cremoris were determined. Unrelated strains yielded markedly different patterns of digestion products. Bacteriophage-resistant derivatives of four strains, generated by a method analogous to that used regularly in some cheese factories, yielded patterns that were identical or almost identical to that of the parent strain. It is proposed that a 16-h PFGE run with a pulse time increasing linearly from 1 to 20 s, which separates fragments between 50 and 240 kilobase pairs (kbp) and produces a pattern containing around 15 bands, can be used as a reliable procedure for strain identification in the lactococci. SmaI digests of 24 of the strains were analyzed by PFGE at three different pulse times to determine accurately the sizes of fragments bigger than 8 kbp. The sum of the sizes of all of the fragments in the digest of a strain provided an estimate of the genome size of the strain. For all the strains analyzed, this estimate was within the range of 2.0 to 2.7 Mbp, with no apparent difference between L. lactis subsp. lactis, L. lactis subsp. lactis biovar diacetylactis and L. lactis subsp. cremoris strains. PMID- 16348319 TI - Enhancing the Viability of Lactobacillus plantarum Inoculum by Immobilizing the Cells in Calcium-Alginate Beads Incorporating Cryoprotectants. AB - Many literature reports have cited the importance of the rehydration conditions of lyophilized cultures in determining viability. The rate of rehydration and the volume of fluid used have been identified as two important factors. One possible means of controlling these is by immobilizing the cells before lyophilization within a gel matrix in which the subsequent rehydration rate and fluid volume would be controlled by the properties of the gel. In this study Lactobacillus plantarum was immobilized and lyophilized in Ca-alginate beads in which 1 M glycerol or 0.75 M adonitol with skim milk were incorporated as a cryoprotectant. The properties of these Ca-alginate beads were examined before and after lyophilization and rehydration. The beads incorporating glycerol were smaller and stronger than those with adonitol. After lyophilization, size decreased and strength increased but to a greater extent in the beads with glycerol, indicating that the microenvironment within the two bead types was probably different. The protective effect of the bead microenvironment on immobilized L. plantarum was also examined. Lyophilization and rehydration within the alginate beads with either polyol yielded higher survival rates than that attained with free cell cultures during rehydration in optimal or suboptimal conditions. During rehydration under suboptimal conditions, the immobilized cell survival was greatest when 0.75 M adonitol was the incorporated cryoprotectant. PMID- 16348320 TI - Biosynthesis of Poly-beta-Hydroxyalkanoates from Pentoses by Pseudomonas pseudoflava. AB - The potential of Pseudomonas pseudoflava to produce poly-beta-hydroxyalkanoates (PHAs) from pentoses was studied. This organism was able to use a hydrolysate from the hemicellulosic fraction of poplar wood as a carbon and energy source for its growth. However, in batch cultures, growth was inhibited completely at hydrolysate concentrations higher than 30% (vol/vol). When P. pseudoflava was grown on the major sugars present in hemicelluloses in batch cultures, poly-beta hydroxybutyric acid (PHB) accumulated when glucose, xylose, or arabinose was the sole carbon source, with the final PHB content varying from 17% (wt/wt) of the biomass dry weight on arabinose to 22% (wt/wt) of the biomass dry weight on glucose and xylose. Specific growth rates were 0.58 h on glucose, 0.13 h on xylose, and 0.10 h on arabinose, while the specific PHB production rates based on total biomass ranged from 0.02 g g h on arabinose to 0.11 g g h on glucose. PHB weight-average molecular weights were 640,000 on arabinose and 1,100,000 on glucose and xylose. The absolute amount of PHB in the cells decreased markedly when nitrogen limitation was relaxed by feeding ammonium sulfate at the end of the PHB accumulation stage of the arabinose and xylose fermentations. Copolymers of beta-hydroxybutyric and beta-hydroxyvaleric acids were produced when propionic acid was added to shake flasks containing 10 g of glucose liter. The beta hydroxyvaleric acid monomer content attained a maximum of 45 mol% when the initial propionic acid concentration was 2 g liter. PMID- 16348321 TI - Mathematical Model of Plasmid Transfer between Strains of Streptomycetes in Soil Microcosms. AB - A mathematical model was developed and used to simulate the long-term dynamics of growth and plasmid transfer in nutrient-limited soil microcosms of Streptomyces lividans TK24 carrying chromosomal resistance to streptomycin, S. lividans 1326; and S. violaceolatus ISP5438. Donor, recipient, and transconjugant survival was modelled by an extension to the Verhulst logistic equation which takes account of nutrient limitation, and plasmid transfer was modelled by a mass action model. Rate parameters were derived from experimental data on the early stages of the development of sterile systems. The model predicted donor, recipient, and transconjugant populations in 2.4-h (0.1-day) steps and was tested against the long-term behavior of the experimental sterile systems and independent experimental data on nonsterile systems. Bacteria were periodically enumerated onto selective media over a 20-day period. The effects of long-term nutrient moisture depletion were correctly predicted. PMID- 16348322 TI - Digestion of barley, maize, and wheat by selected species of ruminal bacteria. AB - Differences in the digestion of barley, maize, and wheat by three major ruminal starch-digesting bacterial species, Streptococcus bovis 26, Ruminobacter amylophilus 50, and Butyrivibrio fibrisolvens A38, were characterized. The rate of starch digestion in all cereal species was greater for S. bovis 26 than for R. amylophilus 50 or B. fibrisolvens A38. Starch digestion by S. bovis 26 was greater in wheat than in barley or maize, whereas starch digestion by R. amylophilus 50 was greater in barley than in maize or wheat. B. fibrisolvens A38 digested the starch in barley and maize to a similar extent but was virtually unable to digest the starch in wheat. The higher ammonia concentration in cultures of B. fibrisolvens A38 when grown on wheat than when grown on barley or maize suggests that B. fibrisolvens A38 utilized wheat protein rather than starch. Scanning electron microscopy revealed that B. fibrisolvens A38 initially colonized cell wall material, while S. bovis 26 randomly colonized the endosperm and R. amylophilus 50 preferentially colonized starch granules. There was subsequent colonization but only superficial digestion of wheat starch granules by B. fibrisolvens A38. Variation in the association between starch and protein within the endosperm of cereal grains contributes to the differential effectiveness with which amylolytic species can utilize cereal starch. PMID- 16348323 TI - Kinetics of the methanogenic fermentation of acetate. AB - Inhibition of the fermentation of acetate to methane and carbon dioxide by acetate was analyzed with an acetate-acclimatized sludge and with Methanosarcina barkeri Fusaro under mesophilic conditions. A second-order substrate inhibition model, q(ch(4) ) = q(m)S/[K(s) + S + (S/K(i))], where S was the concentration of undissociated acetic acid, not ionized acetic acid, could be applicable in both cases. The analysis resulted in substrate saturation constants, K(s), of 4.0 muM for the acclimatized sludge and 104 muM for M. barkeri. The threshold concentrations of undissociated acetic acid when no further acetate utilization was observed were 0.078 muM (pH 7.50) for the acclimatized sludge and 4.43 muM (pH 7.45) for M. barkeri. These kinetic results suggested that the concentration of undissociated acetic acid became a key factor governing the actual threshold acetate concentration for acetate utilization and that the acclimatized sludge in which Methanothrix spp. appeared dominant could utilize acetate better and survive at a lower concentration of undissociated acetic acid than could M. barkeri. PMID- 16348324 TI - Purification and Characterization of an Aspecific Glycoside Hydrolase from the Anaerobic Ruminal Fungus Neocallimastix frontalis. AB - A glycoside hydrolase characterized by beta-fucosidase (EC 3.2.1.38) and beta glucosidase (EC 3.2.1.21) activities was purified from the culture medium of the anaerobic ruminal phycomycete Neocallimastix frontalis grown on 0.5% Avicel. The enzyme had a molecular mass of 120 kilodaltons and a pI of 3.85. Optimal activity against p-nitrophenyl-beta-d-fucoside and p-nitrophenyl-beta-D-glucoside occurred at pH 6.0 and 50 degrees C. The beta-fucosidase and beta-glucosidase activities were stable from pH 6.0 to pH 7.8 and up to 40 degrees C. They were both inhibited by gluconolactone, sodium dodecyl sulfate, p-chloromercuribenzoate, and Hg cation. The enzyme had K(m)s of 0.26 mg/ml for p-nitrophenyl-beta-d-fucoside and 0.08 mg/ml for p-nitrophenyl-beta-d-glucoside. The purified protein also had low beta-galactosidase activity. PMID- 16348325 TI - Carbohydrate Storage in the Entomopathogenic Fungus Beauveria bassiana. AB - The entomopathogenic fungus Beauveria bassiana was grown in 1% (wt/vol) gelatin liquid media singly supplemented with a monosaccharide (glucose or fructose), a disaccharide (maltose or trehalose), a polyol (glycerol, mannitol, or sorbitol), or the amino sugar N-acetyl-d-glucosamine. The relative contributions of the carbohydrate, protein, and water contents in the fungal biomass were determined. Carbohydrates composed 18 to 42% of the mycelial dry weight, and this value was lowest in unsupplemented medium and highest in medium supplemented with glucose, glycerol, or trehalose. Biomass production was highest in liquid cultures supplemented with trehalose. When liquid cultures were grown in medium supplemented with 0 to 1% (wt/vol) glucose, trehalose, or N-acetyl-d-glucosamine, there was an increase in the biomass production and the contribution of carbohydrate to mycelial dry weight. Regardless of the glucose concentration in the culture, water content of the mycelia remained about 77.5% (wt/wt). Mycelial storage carbohydrates were determined by capillary gas chromatography. In gelatin liquid medium supplemented with 1% (wt/vol) glucose, B. bassiana stored glycogen (12.0%, wt/dry wt) and the polyols mannitol (2.2%), erythritol (1.6%), glycerol (0.4%), and arabitol (0.1%). Without glucose, B. bassiana stored glycogen (5.4%), mannitol (0.8%), glycerol (0.6%), and erythritol (0.6%) but not arabitol. To our knowledge, this is the first report of carbohydrate storage in an entomopathogenic fungus, and the results are discussed in relation to other fungi and the potential implications to commercial formulation and insect-fungus interactions. PMID- 16348326 TI - Increased Accumulation of Trehalose in Rhizobia Cultured under 1% Oxygen. AB - The growth of rhizobia under 1% O(2) induced the accumulation of alpha,alpha trehalose, and the effect of low O(2) was independent of medium composition and Rhizobium species. Trehalose concentration in cells declined rapidly when microaerobic cultures were supplied with 21% O(2). Trehalose formation in nodules may be induced by the microaerobic environment. PMID- 16348327 TI - 3-Phenylpropanoic Acid Improves the Affinity of Ruminococcus albus for Cellulose in Continuous Culture. AB - A continuous-culture device, adapted for use with solid substrates, was used to evaluate the effects of 3-phenylpropanoic acid (PPA) upon the ability of the South African strain Ruminococcus albus Ce63 to ferment cellulose. Steady states of fermentation were established with a dilution rate of 0.17 h, and the extent and volumetric rates of cellulose fermentation were determined over four consecutive days. When the growth medium contained no additions (control), 25 muM phenylacetate alone, 25 muM PPA alone, or 25 muM each of phenylacetate and PPA, the extent of cellulose hydrolysis was determined to be 41.1, 35.7, 90.2, and 86.9%, respectively, and the volumetric rate of cellulose hydrolysis was 103.0, 97.9, 215.5, and 230.4 mg liter h, respectively. To evaluate the effect of PPA availability on affinity for cellulose, the values for dilution rate and extent of cellulose hydrolysis were used in combination with values for maximum specific growth rate determined from previous studies of growth rates and kinetics of cellulose hydrolysis. The findings support the contention that PPA maintains a competitive advantage for R. albus when grown in a dynamic, fiber-rich environment. PMID- 16348328 TI - Nutritional Features of Syntrophomonas wolfei. AB - Syntrophomonas wolfei subsp. wolfei grew poorly in a defined medium with crotonate as the energy source in the absence of rumen fluid. Thiamine, lipoic acid, biotin, cyanocobalamin, and para-aminobenzoic acid were required for growth comparable to that obtained with the rumen fluid-based medium. Iron and cobalt were also required for the growth of S. wolfei in the chemically defined medium. PMID- 16348329 TI - Evidence that Cellulolysis by an Anaerobic Ruminal Fungus Is Catabolite Regulated by Glucose, Cellobiose, and Soluble Starch. AB - A Piromyces-like ruminal fungus was used to study preferential carbohydrate utilization of [U-C]cellulose, both alone and in combination with several soluble sugars. For cells grown on cellulose alone, cellulolytic activity was immediate and, initially, greater than that observed in the presence of added carbohydrate. Cellulolytic activity remained minimal in cultures containing cellulose plus glucose or cellobiose until the soluble sugar was depleted. Soluble starch also regulated cellulose activity but to a lesser extent. The results presented suggest that some fungal cellulases are susceptible to catabolite regulatory mechanisms. PMID- 16348330 TI - Cell Surface Characteristics of Bacteriophage-Resistant Lactococcus lactis subsp. cremoris SK110 and Its Bacteriophage-Sensitive Variant SK112. AB - Several cell surface characteristics of bacteriophage-resistant Lactococcus lactis subsp. cremoris SK110 were compared with those of its phage-sensitive derivative SK112. After centrifugation, SK110 cells resisted suspension more strongly than SK112 cells. SK112 was more negatively charged and had a more hydrophobic cell surface than SK110. Furthermore, SK112 was agglutinated in the presence of concanavalin A, whereas SK110 was not. The opposite was observed upon incubation of cells of either strain with a lectin from Ricinus communis. A mild alkali treatment decreased the differences in the cell surface characteristics of the two strains remarkably. PMID- 16348331 TI - Effects of Partial O(2) Pressure, Partial CO(2) Pressure, and Agitation on Growth Kinetics of Azospirillum lipoferum under Fermentor Conditions. AB - Azospirillum lipoferum crt1 was grown in batch cultures under standard conditions at 85% saturation of dissolved oxygen (DO) and 30-g/liter glucose concentrations. Kinetic studies revealed nutritional limitations of growth and the presence of an initial lag phase prior to consumption of glucose. The influences of various gaseous environments and shear stress on growth, i.e., various conditions of agitation-aeration, were characterized. Faster growth in the first stages of the culture and shorter duration of the lag phase were observed at DO concentrations of <30% saturation. The possible influences of dissolved CO(2) concentration or shear stress or both were discounted, and we confirmed the detrimental effect of high DO levels (up to 80% saturation) and the favorable influence of low DO concentrations (lower than 30% saturation) on growth. It was concluded that the gaseous environment, i.e., the DO concentration, needs to be considered as an operating parameter and be accurately controlled to ensure optimal growth of Azospirillum cells. PMID- 16348332 TI - Double-antibody sandwich enzyme-linked immunosorbent assay for cellobiohydrolase I. AB - A double-antibody sandwich enzyme-linked immunosorbent assay was developed for quantifying cellobiohydrolase I (CBH I) in crude preparations of the cellulase complex from Trichoderma reesei. The other enzymes (endoglucanase and beta glucosidase) in this complex and other ingredients in culture broth did not interfere with this assay. The antibody configuration that resulted in the highest specificity for the assay of CBH I employed a monoclonal antibody to coat wells in polystyrene plates and peroxidase-labeled polyclonal antibody to detect cellobiohydrolase bound to the immobilized monoclonal antibody. Previously, procedures have not been available for the direct assay of CBH I activity in the presence of the other enzymes in the complex, and current indirect procedures are cumbersome and inaccurate. The direct procedure described here is highly specific for CBH I and useful for quantifying this enzyme in the range of 0.1 to 0.8 mug/ml. PMID- 16348333 TI - Effects of Hg, CH(3)-Hg, and Temperature on the Expression of Mercury Resistance Genes in Environmental Bacteria. AB - Twenty different bacterial isolates obtained from a mercury-contaminated site in Oak Ridge, Tenn., were grown on plate count agar amended with 25 mug of Hg or 3 mug of CH(3)-Hg (R-Hg) per ml. The total cellular RNA was extracted from each isolate by an acid-guanidine-thiocyanate-phenol-chloroform method. The transcripts of merA and merB were detected and quantitated by Northern (RNA) hybridization. A qualitative assay of mercuric reductase was used to confirm the enzyme activity. Low temperature (4 degrees C) with the presence of Hg (25 mug/ml) significantly increased the net merA transcripts of mid-log-phase cells of six environmental isolates. The net merA transcript production by 18 of the isolates increased when they were grown on 50% plate count broth with 15 mug of Hg per ml, but only 8 isolates showed increased production of merB transcripts. The MICs of Hg and R-Hg for 10 methyl mercury-resistant isolates ranged from 45 to 110 mug of Hg and 0.6 to 4.5 mug of R-Hg per ml. R-Hg was able to induce the expression of merB in 70% of methyl mercury-resistant strains. PMID- 16348334 TI - Selective Medium for Isolation of Mycoleptodiscus terrestris from Soil Sediments of Aquatic Environments. AB - A selective medium was developed for the dilution plate isolation of Mycoleptodiscus terrestris from natural soils and sediments from aquatic environments. The ingredients per liter of the selective medium are as follows: KH(2)PO(4), 0.5 g; MgSO(4) 7H(2)O, 0.5 g; dextrose, 10.0 g; peptone, 5.0 g; chloramphenicol, 0.25 g; rose bengal, 50 mg; oxgall, 5.0 g; Terraclor (pentachloronitrobenzene, 75% active ingredients), 0.5 g; agar, 15.0 g. After autoclaving, the following ingredients were aseptically added: sorbic acid (0.7% autoclave-sterilized aqueous solution), 5.0 ml; Subdue (25.1% emulsion of metalaxyl), 0.5 ml; Truban (40.7% suspension of etridiazol), 0.05 ml. The colony restrictive properties of this medium enabled its use in the drop plate method, originally developed for viable counts of bacteria. Alfalfa sprouts as baits were not suitable for quantitative recovery of the fungus, although 5% of alfalfa sprouts were infected with M. terrestris when incubated on soil containing 1.5 x 10 CFU/g. PMID- 16348335 TI - Isolation and Identification of the Coal-Solubilizing Agent Produced by Trametes versicolor. AB - Low-ranked coals were dissolved by using cell extracts derived from liquid cultures of Trametes versicolor. The coal-solubilizing agent (CSA) was separated from the broth components by a multistep isolation procedure including reverse phase high-pressure liquid chromatography, size exclusion chromatography, ethanol fractionation, and recrystallization. Staircase voltammetry was used to show that two CSA moieties can coordinate to aqueous copper(II) ion. A molecular weight determination (using amperometry) gave an apparent molecular weight of 1.3(4) x 10 g/mol +/- 8%. Nuclear magnetic resonance indicated that all protons on CSA are exchangeable in D(2)O and that there is only one type of carbon in CSA. The infrared spectrum of recrystallized CSA is identical to that of ammonium oxalate, and X-ray studies confirmed the crystal structure and composition of CSA to be that of ammonium oxalate monohydrate. The equivalent weight of the coal in solution, when the coal was dissolved by ammonium oxalate, is 7,940 g of coal per mol of iron present in the coal. PMID- 16348336 TI - Dimethyl sulfide production from dimethylsulfoniopropionate in coastal seawater samples and bacterial cultures. AB - Dimethyl sulfide (DMS) was produced immediately after the addition of 0.1 to 2 muM beta-dimethylsulfonio-propionate (DMSP) to coastal seawater samples. Azide had little effect on the initial rate of DMS production from 0.5 muM added DMSP, but decreased the rate of production after 6 h. Filtration of water samples through membrane filters (pore size, 0.2 mum) greatly reduced DMS production for approximately 10 h, after which time DMS production resumed at a high rate. Autoclaving completely eliminated the production of DMS. The antibiotics chloramphenicol, tetracycline, kanamycin, and vancomycin all had little effect on the accumulation of DMS over the first few hours of incubation, but produced significant inhibition thereafter. The effects of individual antibiotics were additive. Chloroform over a range of concentrations (0.25 to 1.25 mM) had no effects on DMS production. Similarly, organic amendments, including acrylate, glucose, protein, and starch, did not affect DMS accumulation from DMSP. Acrylate, a product of the enzymatic cleavage of DMSP, was metabolized in seawater samples, and two strains of bacteria were isolated with this compound as the growth substrate. These bacteria produced DMS from DMSP. The sensitivity to inhibitors with respect to growth and DMSP-lyase activity varied from strain to strain. These results illustrate the significant potential for microbial conversion of dissolved DMSP to DMS in coastal seawater. PMID- 16348337 TI - Single-Strain versus Multistrain Inoculation: Effect of Soil Mineral N Availability on Rhizobial Strain Effectiveness and Competition for Nodulation on Chick-Pea, Soybean, and Dry Bean. AB - The nitrogen-fixing effectiveness of multistrain inoculants was found to be determined by both the effectiveness of the component strains and the percentage of the nodules occupied by them. Multistrain formulations were always either as good as the most effective single-strain inoculant or intermediate between the most and the least effective. The percentage of nodules occupied and the amount of nitrogen fixed by the component strains of a multistrain inoculant showed highly significant linear correlation. The availability of soil N had a significant influence on the nitrogen fixation potential of each strain. The mineral N status of the soil was clearly a significant factor in affecting the competition pattern of Rhizobium loti (chick-pea) and Bradyrhizobium japonicum strains. Differences between the effectiveness of strains were masked under conditions of soil N availability. However, when soil N was immobilized with sugarcane bagasse, the differences became significant. In the chick-pea system, R. loti TAL 1148 (Nit 27A8) was the most effective but not the most competitive of the three strains used. In the soybean and dry bean systems, B. japonicum TAL 102 (USDA 110) and R. leguminosarum bv. phaseoli TAL 182, respectively, were consistently the most effective and, more often than not, the most competitive of the strains used for each species. PMID- 16348338 TI - Susceptibility to Heavy Metals and Characterization of Heterotrophic Bacteria Isolated from Two Hydrothermal Vent Polychaete Annelids, Alvinella pompejana and Alvinella caudata. AB - Specimens of alvinellid polychaetes and their tubes were collected in the Parigo hydrothermal vent field on the East Pacific Rise (13 degrees N) in October and November 1987. Heterotrophic bacterial strains were isolated on metal-amended media from the tube and dorsal integument of one specimen of Alvinella pompejana, from the dorsal integument of another from the whole integument of a specimen of Alvinella caudata, and from undetermined alvinellid tubes. The strains were characterized and tested for susceptibility to five heavy metals by using a microdilution method for MIC determinations. All strains were gram-negative rods. Most of them were characterized by the ability to degrade Tween 80 and gelatin and to produce hydrogen sulfide from cysteine. Numerous strains, from all sample origins, displayed resistance to cadmium, zinc, arsenate, and silver and tolerated high amounts of copper. Metal resistance was exhibited by 92.3% of the total isolates. The occurrence of multiply resistant bacteria may demonstrate an adaptation of alvinellid-associated microflora to the general enrichment of metals in the hydrothermal vent environment. PMID- 16348339 TI - Isolation and Characterization of Pichia heedii Mutants Defective in Xylose Uptake. AB - To investigate the role of xylose uptake in xylose metabolism in yeasts, we isolated a series of mutated strains of the yeast Pichia heedii which are defective in xylose utilization. Four of these demonstrated defects in xylose uptake. Overlaps between the functional or regulatory mechanisms for glucose and xylose uptake may exist in this yeast since some of the mutants defective in xylose uptake were also defective in glucose transport. None of the mutants were defective in xylose reductase or xylitol dehydrogenase activities. PMID- 16348340 TI - Degradation of the Ferric Chelate of EDTA by a Pure Culture of an Agrobacterium sp. AB - A pure culture of an Agrobacterium sp. (deposited as ATCC 55002) that mineralizes the ferric chelate of EDTA (ferric-EDTA) was isolated by selective enrichment from a treatment facility receiving industrial waste containing ferric-EDTA. The isolate grew on ferric-EDTA as the sole carbon source at concentrations exceeding 100 mM. As the degradation proceeded, carbon dioxide, ammonia, and an unidentified metabolite(s) were produced; the pH increased, and iron was precipitated from solution. The maximum rate of degradation observed with sodium ferric-EDTA as the substrate was 24 mM/day. At a substrate concentration of 35 mM, 90% of the substrate was degraded in 3 days and 70% of the associated chemical oxygen demand was removed from solution. Less than 15% of the carbon initially present was incorporated into the cell mass. Significant growth of this strain was not observed with uncomplexed EDTA as the sole carbon source at comparable concentrations; however, the ferric chelate of propylenediaminetetraacetic acid (ferric-PDTA) did support growth. PMID- 16348341 TI - Accumulation of a Polyhydroxyalkanoate Containing Primarily 3-Hydroxydecanoate from Simple Carbohydrate Substrates by Pseudomonas sp. Strain NCIMB 40135. AB - A number of Pseudomonas species have been identified which accumulate a polyhydroxyalkanoate containing mainly 3-hydroxydecanoate monomers from sodium gluconate as the sole carbon source. One of these, Pseudomonas sp. strain NCIMB 40135, was further investigated and shown to accumulate such a polyhydroxyalkanoate from a wide range of carbon sources (C(2) to C(6)); however, when supplied with octanoic acid it produced a polyhydroxyalkanoate containing mainly 3-hydroxyoctanoate monomers. Polymer synthesis occurred in batch culture after cessation of growth due to exhaustion of nitrogen. In continuous culture under nitrogen limitation up to 16.9% (wt/wt) polyhydroxyalkanoate was synthesized from glucose as the carbon source. The monomer units are mainly of the R-(-) configuration. Nuclear magnetic resonance studies confirmed the composition of the polymer. Differential scanning calorimetry suggested that the solvent-extracted polymer contained a significant proportion of crystalline material. The weight-average molecular weight of the polymer from glucose-grown cells was 143,000. PMID- 16348342 TI - Fast-growing, aerobic, heterotrophic bacteria from the rhizosphere of young sugar beet plants. AB - Fast-growing, aerobic, heterotrophic bacteria from the root surface of young sugar beet plants were inventoried. Isolation of the most abundant bacteria from the root surface of each of 1,100 plants between the second and tenth leaf stage yielded 5,600 isolates. These plants originated from different fields in Belgium and Spain. All isolates were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis of total cellular proteins. Comparison of protein fingerprints allowed us to inventory the bacteria of individual plants of different fields or leaf stages and to analyze the composition and variability of the rhizobacterial population of young sugar beet plants. Each field harbored a specific population of bacteria which showed a highly hierarchic structure. A small number of bacteria occurring frequently at high densities dominated in each field. The major bacteria were identified as Pseudomonas fluorescens, Xanthomonas maltophilia, Pseudomonas paucimobilis, and Phyllobacterium sp. The former three species showed a high genetic variability as they were represented by different protein fingerprint types on the same or different fields or leaf stages. Twinspan analysis and relative abundance plots showed that the structure and composition of the bacterial populations varied strongly over time. Pseudomonads were typically early colonizers which were later replaced by X. maltophilia or Phyllobacterium sp. PMID- 16348343 TI - Effect of Oxygenation on Xylose Fermentation by Pichia stipitis. AB - The effect of oxygen limitation on xylose fermentation by Pichia stipitis (CBS 6054) was investigated in continuous culture. The maximum specific ethanol productivity (0.20 g of ethanol g dry weight h) and ethanol yield (0.48 g/g) was reached at an oxygen transfer rate below 1 mmol/liter per h. In the studied range of oxygenation, the xylose reductase (EC 1.1.1.21) and xylitol dehydrogenase (EC 1.1.1.9) activities were constant as well as the ratio between the NADPH and NADH activities of xylose reductase. No xylitol production was found. The pyruvate decarboxylase (EC 4.1.1.1) activity increased and the malate dehydrogenase (EC 1.1.1.37) activity decreased with decreasing oxygenation. With decreasing oxygenation, the intracellular intermediary metabolites sedoheptulose 7 phosphate, glucose 6-phosphate, fructose 1,6-diphosphate, and malate accumulated slightly while pyruvate decreased. The ratio of the xylose uptake rate under aerobic conditions, in contrast to that under anaerobic assay conditions, increased with increasing oxygenation in the culture. The results are discussed in relation to the energy level in the cell, the redox balance, and the mitochondrial function. PMID- 16348344 TI - Mixotrophic and Autotrophic Growth of Thiobacillus acidophilus on Glucose and Thiosulfate. AB - Mixotrophic growth of the facultatively autotrophic acidophile Thiobacillus acidophilus on mixtures of glucose and thiosulfate or tetrathionate was studied in substrate-limited chemostat cultures. Growth yields in mixotrophic cultures were higher than the sum of the heterotrophic and autotrophic growth yields. Pulse experiments with thiosulfate indicated that tetrathionate is an intermediate during thiosulfate oxidation by cell suspensions of T. acidophilus. From mixotrophic growth studies, the energetic value of thiosulfate and tetrathionate redox equivalents was estimated to be 50% of that of redox equivalents derived from glucose oxidation. Ribulose 1,5-bisphosphate carboxylase (RuBPCase) activities in cell extracts and rates of sulfur compound oxidation by cell suspensions increased with increasing thiosulfate/glucose ratios in the influent medium of the mixotrophic cultures. Significant RuBPCase and sulfur compound-oxidizing activities were detected in heterotrophically grown T. acidophilus. Polyhedral inclusion bodies (carboxysomes) could be observed at low frequencies in thin sections of cells grown in heterotrophic, glucose-limited chemostat cultures. Highest RuBPCase activities and carboxysome abundancy were observed in cells from autotrophic, CO(2)-limited chemostat cultures. The maximum growth rate at which thiosulfate was still completely oxidized was increased when glucose was utilized simultaneously. This, together with the fact that even during heterotrophic growth the organism exhibited significant activities of enzymes involved in autotrophic metabolism, indicates that T. acidophilus is well adapted to a mixotrophic lifestyle. In this respect, T. acidophilus may have a competitive advantage over autotrophic acidophiles with respect to the sulfur compound oxidation in environments in which organic compounds are present. PMID- 16348345 TI - Inverse Diauxy in the Yeast Hansenula anomala: Mutants Derepressed for Malic Acid Utilization in the Presence of Glucose. AB - Utilization of l-malic acid by yeast strain Hansenula anomala IGC 4380 is subject to glucose repression. Derepressed mutants were obtained with UV light by use of the nonmetabolizable glucose analog 2-deoxyglucose as a selective agent. Three mutant strains degraded l-malic acid in the presence of up to 30% (wt/vol) glucose and are of potential interest for the biological deacidification of grape must. The mutant strains, as compared with the parent strain, displayed inverse diauxy in glucose-malate medium, glucose being metabolized only after malate consumption had been completed. PMID- 16348346 TI - Rapid methane oxidation in a landfill cover soil. AB - Methane oxidation rates observed in a topsoil covering a retired landfill are the highest reported (45 g m day) for any environment. This microbial community had the capacity to rapidly oxidize CH(4) at concentrations ranging from <1 ppm (microliters per liter) (first-order rate constant [k] = -0.54 h) to >10 ppm (k = -2.37 h). The physiological characteristics of a methanotroph isolated from the soil (characteristics determined in aqueous medium) and the natural population, however, were similar to those of other natural populations and cultures: the Q(10) and optimum temperature were 1.9 and 31 degrees C, respectively, the apparent half-saturation constant was 2.5 to 9.3 muM, and 19 to 69% of oxidized CH(4) was assimilated into biomass. The CH(4) oxidation rate of this soil under waterlogged (41% [wt/vol] H(2)O) conditions, 6.1 mg liter day, was near rates reported for lake sediment and much lower than the rate of 116 mg liter day in the same soil under moist (11% H(2)O) conditions. Since there are no large physiological differences between this microbial community and other CH(4) oxidizers, we attribute the high CH(4) oxidation rate in moist soil to enhanced CH(4) transport to the microorganisms; gas-phase molecular diffusion is 10-fold faster than aqueous diffusion. These high CH(4) oxidation rates in moist soil have implications that are important in global climate change. Soil CH(4) oxidation could become a negative feedback to atmospheric CH(4) increases (and warming) in areas that are presently waterlogged but are projected to undergo a reduction in summer soil moisture. PMID- 16348348 TI - Carbon Concentration and Carbon-to-Nitrogen Ratio Influence Submerged-Culture Conidiation by the Potential Bioherbicide Colletotrichum truncatum NRRL 13737. AB - We assessed the influence of various carbon concentrations and carbon-to-nitrogen (C:N) ratios on Colletotrichum truncatum NRRL 13737 conidium formation in submerged cultures grown in a basal salts medium containing various amounts of glucose and Casamino Acids. Under the nutritional conditions tested, the highest conidium concentrations were produced in media with carbon concentrations of 4.0 to 15.3 g/liter. High carbon concentrations (20.4 to 40.8 g/liter) inhibited sporulation and enhanced the formation of microsclerotiumlike hyphal masses. At all the carbon concentrations tested, a culture grown in a medium with a C:N ratio of 15:1 produced more conidia than cultures grown in media with C:N ratios of 40:1 or 5:1. While glucose exhaustion was often coincident with conidium formation, cultures containing residual glucose sporulated and those with high carbon concentrations (>25 g/liter) exhausted glucose without sporulation. Nitrogen source studies showed that the levels of C. truncatum NRRL 13737 conidiation were similar for all protein hydrolysates tested. Reduced conidiation occurred when amino acid and inorganic nitrogen sources were used. Of the nine carbon sources evaluated, acetate as the sole carbon source resulted in the lowest level of sporulation. PMID- 16348347 TI - Evidence for a Plasmid-Linked Restriction-Modification System in Lactobacillus helveticus. AB - The presence of a restriction-modification (R/M) system against two bacteriophages, 328-B1 and hv, was demonstrated in three Lactobacillus helveticus strains, CNRZ 1094, CNRZ 1095, and CNRZ 1096. In addition, the burst size of phage 328-B1 in the three restrictive strains CNRZ 1094, CNRZ 1095, and CNRZ 1096 was reduced with respect to the values obtained in its propagating strain, CNRZ 328. Heating at 60 degrees C did not inactivate the R/M system. Nonrestrictive variants from CNRZ 1094 were easily obtained under several culture conditions, but treatment with novobiocin at 42 degrees C followed by storage at -20 degrees C resulted in drastic elimination of the R/M phenotype from all clones tested. Electrophoretic analysis of CNRZ 1094 nonrestrictive variants revealed the concomitant loss of a 34-kb plasmid. Four EcoRI fragments from the 34-kb plasmid were cloned in the Escherichia coli vector pACYC184. The use of one or several of these fragments as probes confirmed the plasmidic location of the genes responsible for the R/M system. These probes also showed the presence of R/M plasmids in the two other restrictive strains, CNRZ 1095 and CNRZ 1096. Lactose fermenting ability and/or proteolytic capacity was not linked to the 34-kb plasmid. PMID- 16348349 TI - Bacteriochlorophyll and Photosynthetic Reaction Centers in Rhizobium Strain BTAi 1. AB - Rhizobium strain BTAi 1, which nodulates both stems and roots of Aeschynomene indica L., formed bacteriochlorophyll and photosynthetic reaction centers resembling those of purple photosynthetic bacteria when grown aerobically ex planta under a light-dark cycle. Bacteriochlorophyll formation was not observed under continuous dark or light growth conditions. The amount of pigment formed was similar to that previously found in aerobic photosynthetic bacteria. Stem nodules appear to fix nitrogen photosynthetically, as illumination of A. indica stem nodules with near-infrared light resulted in an enhanced rate of acetylene reduction. Near-infrared light did not enhance acetylene reduction when either A. indica or soybean root nodules were illuminated. The BTAi 1 isolate can be differentiated from members of the family Rhodospirillaceae by several criteria. PMID- 16348350 TI - Chloromethane, Methyl Donor in Veratryl Alcohol Biosynthesis in Phanerochaete chrysosporium and Other Lignin-Degrading Fungi. AB - Chloromethane, a gaseous natural product implicated in methylation processes in Phellinus pomaceus, has been shown to act as methyl donor in veratryl alcohol biosynthesis in the lignin-degrading fungi Phanerochaete chrysosporium, Phlebia radiata, and Coriolus versicolor, none of which released detectable amounts of CH(3)Cl during growth. When P. chrysosporium was grown in a medium containing CH(3)Cl, levels of CH(3) incorporation into the 3- and 4-O-methyl groups of veratryl alcohol were very high and initially similar to those observed when the medium was supplemented with l-[methyl-H(3)]methionine. When CH(3)Cl was added to cultures actively synthesizing veratryl alcohol, incorporation of CH(3) was very rapid, with 81% of veratryl alcohol labeled after 12 h. By contrast, incorporation of CH(3) from l-[methyl-H(3)]methionine was comparatively slow, attaining 10% after 12 h. It is proposed that these lignin-degrading fungi possess a tightly channeled multienzyme system in which CH(3)Cl biosynthesis is closely coupled to CH(3)Cl utilization for methylation of veratryl alcohol precursors. PMID- 16348351 TI - Oxidation of Manganese and Iron by Leptothrix discophora: Use of N,N,N',N' Tetramethyl-p-Phenylenediamine as an Indicator of Metal Oxidation. AB - A new method for the quantification and characterization of manganese-oxidizing activity by spent culture medium of Leptothrix discophora SS-1 was developed. It is based on the formation of the dye Wurster blue from N,N,N',N'-Tetramethyl-p phenylenediamine by oxidized manganese generated in the spent medium. The kinetic parameters thus obtained agreed well with data obtained with other methods. It was also possible to demonstrate iron oxidation by spent culture medium. The kinetics of the process and inhibition by enzyme poisons suggest that iron oxidation is enzymatically catalyzed. Probably two different factors are involved in manganese and iron oxidation. PMID- 16348352 TI - Immobilization of chloroperoxidase on aminopropyl-glass. AB - Chloroperoxidase (CPO) purified from Caldariomyces fumago CMI 89362 was covalently bound to aminopropyl-glass by using a modification of an established method. Acid-washed glass was derivatized by using aminopropyltriethoxysilane, and the enzyme was ionically bound at low ionic strength. Further treatment with glutaraldehyde covalently linked the enzyme to the glass beads in an active form. No elution of bound activity from glass beads could be detected with a variety of washings. The loading of enzyme protein to the glass beads was highest, 100 mg of CPO per g of glass, at high reaction ratios of CPO to glass, but the specific activity of the immobilized enzyme was highest, 36% of theoretical, at low enzyme to-carrier ratios. No differences in the properties of the soluble and immobilized enzymes could be detected by a number of criteria: their pH-activity and pH-stability profiles were similar, as were their thermal stabilities. After five uses, the immobilized enzyme retained full activity between pH 6.0 and 6.7. PMID- 16348353 TI - Ammonium Excretion by an l-Methionine-dl-Sulfoximine-Resistant Mutant of the Rice Field Cyanobacterium Anabaena siamensis. AB - An ammonium-excreting mutant (SS1) of the rice field nitrogen-fixing cyanobacterium Anabaena siamensis was isolated after ethyl methanesulfonate mutagenesis by selection on 500 muM l-methionine-dl-sulfoximine. SS1 grew in the presence and absence of (l)-methionine-dl-sulfoximine at a rate comparable to that of the wild-type strain, with a doubling time of 5.6 h. The rate of ammonium release by SS1 depended on cell density; it peaked at the 12th hour of growth with 8.7 mumol mg of chlorophyll h (at a chlorophyll concentration of 5 mug ml) and slowed down to almost nil at the fourth day of growth. A similar pattern of release by immobilized SS1 was observed between 12 to 20 h after loading alginate beads in packed-bed reactors at the rate of 11.6 mumol mg of chlorophyll h. The rate was later reduced significantly due to the fast growth of SS1 on the substrate. Prolonged release of ammonium at the peak level was achieved only by maintaining SS1 under continuous cultivation at low chlorophyll levels (5 to 7 mug ml). Under these conditions, nitrogen fixation in the mutant was 30% higher than that in its parent and glutamine synthetase activity was less by 50%. Immunoblot analysis revealed that SS1 and its parent have similar quantities of glutamine synthetase protein under ammonium excretion conditions. In addition, a protein with a molecular weight of about 30,000 seems to have been lost, as seen by electrophoretic separation of total proteins from SS1. PMID- 16348354 TI - Effect of Dicarboxylic Acids and Aspergillus oryzae Fermentation Extract on Lactate Uptake by the Ruminal Bacterium Selenomonas ruminantium. AB - The objective of this study was to determine the effects of l-aspartate, fumarate, l-malate, and an Aspergillus oryzae fermentation extract (Amaferm) on growth on lactate as well as lactate uptake by Selenomonas ruminantium HD4. Growth of S. ruminantium in medium that contained 2 g of dl-lactate per liter was stimulated approximately twofold by 10 mM l-aspartate, fumarate, or l-malate after 24 h. Both l-aspartate and fumarate increased lactate uptake over 4-fold, while l-malate stimulated uptake over 10-fold. Amaferm enhanced lactate uptake at all concentrations tested (0.5 to 50 g/liter), and the 10-g/liter level increased uptake over 12-fold. A filter-sterilized Amaferm filtrate increased lactate uptake over sevenfold, and growth on lactate was stimulated over twofold by either 2 or 5% (vol/vol) Amaferm filtrate. The Amaferm filtrate also increased the production of acetate, propionate, total volatile fatty acids, and Y(lactate) from lactate-grown cells. Since the increase in propionate production was greater relative to acetate, a decrease in the acetate:propionate ratio was observed. The concentration of l-malate in the Amaferm filtrate was 1.45 mM, and it appeared that the l-malate content of Amaferm played a role in the stimulation of growth on lactate as well as lactate uptake by S. ruminantium treated with Amaferm. PMID- 16348355 TI - Sensitivity to and Degradation of Pentachlorophenol by Phanerochaete spp. AB - This research measured mycelial extension rates of selected strains of Phanerochaete chrysorhiza, Phanerochaete laevis, Phanerochaete sanguinea, Phanerochaete filamentosa, Phanerochaete sordida, Inonotus circinatus, and Phanerochaete chrysosporium and the ability of these organisms to tolerate and degrade the wood preservative pentachlorophenol (PCP) in an aqueous medium and in soil. Most of the tested species had mycelial extension rates in the range of P. sordida > P. laevis > P. chrysorhiza = P. sanguinea > I. circinatus = P. filamentosa. There were also significant intraspecific differences in mycelial extension rates. For example, mycelial extension rates among strains of P. sordida ranged from 1.78 to 4.81 cm day. Phanerochaete spp. were very sensitive to PCP. Growth of several species was prevented by the presence of 5 ppm (5 mug/g) PCP. However, P. chrysosporium and P. sordida grew at 25 ppm PCP, albeit at greatly decreased mycelial extension rates. In an aqueous medium, mineralization of PCP by P. sordida 13 (ca. 12% after 30 days) was significantly greater than that by all other tested P. sordida strains and P. chrysosporium. After 64 days, the level of PCP had decreased by 96 and 82% in soil inoculated with P. chrysosporium and P. sordida, respectively. Depletion of PCP by these fungi occurred in a two-stage process. The first stage was characterized by a rapid depletion of PCP that coincided with an accumulation of pentachloroanisole (PCA). At the end of the first stage, ca. 64 and 71% of the PCP was converted to PCA in P. chrysosporium and P. sordida cultures, respectively. In the second stage, levels of PCP and PCA were reduced by 9.6 and 18%, respectively, in soil inoculated with P. chrysosporium and by 3 and 23%, respectively, in soil inoculated with P. sordida. PCA was mineralized by both P. chrysosporium and P. sordida in an aqueous medium. PMID- 16348356 TI - Antiserum to Nitrogenase Generated from an Amplified DNA Fragment from Natural Populations of Trichodesmium spp. AB - A fragment of the nifH gene was amplified from natural populations of Trichodesmium spp. and cloned into a maltose-binding protein (MBP) expression vector. The peptide product of the amplified 359-bp fragment of nifH was cleaved from the fusion protein, purified, and used to generate a specific antibody to the Fe protein of nitrogenase. The antiserum recognized the MBP-nitrogenase fusion protein and the cleaved nif peptide product but not MBP. The antibody cross-reacted with nitrogenase from natural populations of Trichodesmium spp. from the Caribbean Sea and with a cultured isolate from the Kuroshio waters (Trichodesmium sp. strain NIBB1067). The same nifH fragment was amplified, cloned, and sequenced from Trichodesmium sp. strain NIBB1067 and was found to be 98% identical at both the protein and DNA levels to nifH from the Caribbean populations. Three of the six nucleotide differences between the Trichodesmium sp. strain NIBB1067 and the Trichodesmium spp. nifH sequence had also been found in a second sequence from the natural populations, indicating either that there is more than one strain of Trichodesmium sp. in natural assemblages or that there are multiple copies of nifH in the genome. This DNA fragment, which is easily amplified with the polymerase chain reaction, may provide a good indicator of species relatedness without requiring extensive cloning or sequencing. Furthermore, the use of the polymerase chain reaction in combination with a MBP protein fusion vector provides a rapid method for production of highly specific sera, starting with a small amount of DNA. PMID- 16348357 TI - Basis for Diel Variation in Nitrogenase Activity in the Marine Planktonic Cyanobacterium Trichodesmium thiebautii. AB - Natural populations of the nonheterocystous marine cyanobacterium Trichodesmium thiebautii exhibit a diel periodicity in nitrogenase activity (NA). NA "turns on" near dawn and "turns off" near dusk, independent of photic conditions. Chloramphenicol (CAP) and ammonium prevented turn on of NA in T. thiebautii when added to samples collected before dawn but were progressively less effective in inhibiting NA in samples collected later in the morning. In samples collected after turn on, activities declined with time with both CAP and ammonium treatments, with ammonium having a stronger effect. In contrast, CAP added to samples collected in late afternoon prolonged NA, compared with controls, which turned off. Direct analysis of the presence of the Fe protein of nitrogenase in T. thiebautii by using a Western immunoblot procedure found a strong protein band present in samples collected after 0800 h through the late evening but little or no Fe protein in samples collected within the 2 to 4 h preceding dawn. We conclude that the diel cycle of NA in T. thiebautii results from de novo synthesis of nitrogenase each morning and from the inactivation and degradation of nitrogenase in the late afternoon and night. PMID- 16348358 TI - Identification of different agrobacterium strains isolated from the same forest nursery. AB - Several Agrobacterium strains isolated from the same forest nursery from 1982 to 1988 were compared by serological, biochemical, and DNA-DNA hybridization methods. Similarities among strains belonging to biovar 2 were observed by indirect immunofluorescence, whereas biovar 1 strains showed serological heterogeneity. Electrophoretic analysis of bacterial envelope-associated proteins showed that few bands appeared in the strains belonging to biovar 1, whereas many proteins appeared in the case of biovar 2 strains. Chromosomal DNA was analyzed with six random C58 chromosomal fragments. None of the six probes hybridized to the DNA of the two biovar 2 strains. One of the probes gave the same hybridization pattern with all biovar 1 strains, whereas the other probes yielded different patterns. The vir regions were closely related in the different pathogenic strains. The T-DNA and replication regions were less conserved and showed some variations among the strains. PMID- 16348359 TI - Dissimilatory selenate reduction potentials in a diversity of sediment types. AB - We measured potential rates of bacterial dissimilatory reduction of SeO(4) to Se in a diversity of sediment types, with salinities ranging from freshwater (salinity = 1 g/liter) to hypersaline (salinity = 320 g/liter and with pH values ranging from 7.1 to 9.8. Significant biological selenate reduction occurred in all samples with salinities from 1 to 250 g/liter but not in samples with a salinity of 320 g/liter. Potential selenate reduction rates (25 nmol of SeO(4) per ml of sediment added with isotope) ranged from 0.07 to 22 mumol of SeO(4) reduced liter h. Activity followed Michaelis-Menten kinetics in relation to SeO(4) concentration (K(m) of selenate = 7.9 to 720 muM). There was no linear correlation between potential rates of SeO(4) reduction and salinity, pH, concentrations of total Se, porosity, or organic carbon in the sediments. However, potential selenate reduction was correlated with apparent K(m) for selenate and with potential rates of denitrification (r = 0.92 and 0.81, respectively). NO(3), NO(2), MoO(4), and WO(4) inhibited selenate reduction activity to different extents in sediments from both Hunter Drain and Massie Slough, Nev. Sulfate partially inhibited activity in sediment from freshwater (salinity = 1 g/liter) Massie Slough samples but not from the saline (salinity = 60 g/liter) Hunter Drain samples. We conclude that dissimilatory selenate reduction in sediments is widespread in nature. In addition, in situ selenate reduction is a first-order reaction, because the ambient concentrations of selenium oxyanions in the sediments were orders of magnitude less than their K(m)s. PMID- 16348360 TI - Catalase and superoxide dismutase of root-colonizing saprophytic fluorescent pseudomonads. AB - Root-colonizing, saprophytic fluorescent pseudomonads of the Pseudomonas putida P. fluorescens group express similar levels of catalase and superoxide dismutase activities during growth on a sucrose- and amino acid-rich medium. Increased specific activities of catalase but not superoxide dismutase were observed during growth of these bacteria on components washed from root surfaces. The specific activities of both enzymes were also regulated during contact of these bacteria with intact bean roots. Increased superoxide dismutase and decreased catalase activities were observed rapidly, by 10 min upon inoculation of cells onto intact bean roots. Catalase specific activity increased with time to peak at 12 h before declining. By 48 h, the cells displayed this low catalase but maintained high superoxide dismutase specific activities. Catalase with a low specific activity and a high superoxide dismutase activity also were present in extracts of cells obtained from 7-day-old roots colonized from inoculum applied to seed. This specific activity of superoxide dismutase of root-contacted cells was about fourfold-higher in comparison to cells grown on rich medium, whereas the specific activity for catalase was reduced about fivefold. A single catalase isozyme, isozyme A, and one isozyme of superoxide dismutase, isozyme 1, were detected during growth of the bacteria on root surface components and during exposure of cells to intact bean roots for 1 h. An additional catalase, isozyme B, was detected from bacteria after exposure to the intact bean roots for 12 h. Catalase isozyme A and superoxide dismutase isozyme 1 were located in the cytoplasm and catalase band B was located in the membrane of P. putida. PMID- 16348361 TI - Characterization and reclassification of yeasts used for biological control of postharvest diseases of fruits and vegetables. AB - In previous studies workers have shown that three yeast strains (strains US-7, 82, and 101) have biological control activity against various postharvest fungal pathogens of fruits and vegetables, including Penicillium rots of apples and citrus and Botrytis rot of apples. In these reports the researchers have described these strains as Debaryomyces hansenii (anamorph, Candida famata) or Candida sp. strains. In this study we performed additional physiological, DNA reassociation, and mannan characterization tests that clearly established a new taxonomic classification for these strains, Candida guilliermondii. We also propose amendment of the physiological test profile in the taxonomic description of C. guilliermondii. PMID- 16348362 TI - Stereospecific reduction of ethyl 2' -ketopantothenate to ethyl d-(+) pantothenate with microbial cells as a catalyst. AB - A novel enzymatic process for the synthesis of D-(+)-pantothenic acid through the asymmetric reduction of the 2' -ketopantothenate ester is described. Candida macedoniensis AKU 4588 was found to convert ethyl 2' -ketopantothenate (80 mg/ml) almost specifically to ethyl D-(+)-pantothenate (>98% enantiomeric excess), with a molar yield of 97.2%. PMID- 16348363 TI - Ecophysiology of Bacteriophage S5100 Infecting Halobacterium cutirubrum. AB - Increasing salinity reduces burst size and increases the latent period of infection of Halobacterium cutirubrum by lytic bacteriophage S5100. Cells become reversibly and persistently infected at saturation-level concentrations of NaCl. We propose that high salinity provides a natural refuge for sensitive host bacteria and that phage S5100 acts as a scavenger, proliferating when host viability is threatened by dilution of the environment. PMID- 16348364 TI - Selective Medium for Enumeration of Tannin-Protein Complex-Degrading Streptococcus spp. in Feces of Koalas. AB - A selective agar plate medium (tannin-treated brain heart infusion agar supplemented with colistin-oxolinic acid) was developed to enumerate tannin protein complex-degrading Streptococcus bovis in the feces of koalas. This medium was successfully used to enumerate strains from fecal samples but failed to enumerate those from pure cultures. PMID- 16348365 TI - Degradation of Alpha-, Beta-, and Gamma-Hexachlorocyclohexane by a Soil Bacterium under Aerobic Conditions. AB - A Pseudomonas sp., isolated from sugarcane rhizosphere soil, readily metabolized not only alpha and gamma isomers of hexachlorocyclohexane, but also the thermodynamically more stable beta isomer, under aerobic conditions. Bacterial degradation of the three isomers led to the accumulation of a transitory metabolite and eventual release of covalently linked chlorine as chloride in stoichiometric amounts. PMID- 16348366 TI - Limited Multiplication of Symbiotic Cyanobacteria of Azolla spp. on Artificial Media. AB - We examined various media and conditions to isolate symbiotic cyanobacteria from the leaf cavities of Azolla spp. Cyanobacteria survived and multiplied to a limited extent on a medium with fructose, Casamino Acids, yeast extract, and NaNO(3) under 1% O(2). These cyanobacteria were antigenically identical to the endosymbionts. PMID- 16348367 TI - Latex and vinyl nonsterile examination gloves: status report on laboratory evaluation of defects by physical and biological methods. AB - [This corrects the article on p. 1627 in vol. 56.]. PMID- 16348368 TI - DNA, a Possible Site of Action of Aluminum in Rhizobium spp. AB - Al was found to penetrate the cell envelopes of both sensitive and tolerant Rhizobium strains and bind to DNA in vivo. Despite causing a reduction in viability, Al stimulated DNA synthesis in the sensitive strain, which suggested that an excision repair mechanism was operating. The Al-stimulated DNA synthesis was reduced by the simultaneous addition of chloramphenicol. In contrast to the sensitive strain, DNA synthesis was unaffected by Al binding to DNA in the tolerant strain. It is proposed that Al enters the cell and binds to the DNA helix, increasing stabilization and preventing successful replication. Different repair mechanisms appear to operate in response to Al in tolerant and sensitive strains. PMID- 16348369 TI - Degradation of adsorbed protein by attached bacteria in relationship to surface hydrophobicity. AB - The relationships among surface energy, adsorbed organic matter, and attached bacterial growth were examined by measuring the degradation of adsorbed ribulose 1,5-bisphosphate carboxylase (a common algal protein) by attached bacteria (Pseudomonas strain S9). We found that surface energy (work of adhesion of water) determined the amount and availability of adsorbed protein and, consequently, the growth of attached bacteria. Percent degradation of adsorbed ribulose-1,5 bisphosphate carboxylase decreased with increasing hydrophobicity of the surface (decreasing work of adhesion). As a result, growth rates of attached bacteria were initially higher on hydrophilic glass than on hydrophobic polyethylene. However, during long (6-h) incubations, growth rates increased with surface hydrophobicity because of increasing amounts of adsorbed protein. Together with previous studies, these results suggest that the number of attached bacteria over time will be a complex function of surface energy. Whereas both protein adsorption and bacterial attachment decrease with increasing surface energy, availability of adsorbed protein and consequently initial bacterial growth rates increase with surface energy. PMID- 16348370 TI - Binding of Dissolved Strontium by Micrococcus luteus. AB - Resting cells of Micrococcus luteus have been shown to remove strontium (Sr) from dilute aqueous solutions of SrCl(2) at pH 7. Loadings of 25 mg of Sr per g of cell dry weight were achieved by cells exposed to a solution containing 50 ppm (mg/liter) of Sr. Sr binding occurred in the absence of nutrients and did not require metabolic activity. Initial binding was quite rapid (<0.5 h), although a slow, spontaneous release of Sr was observed over time. Sr binding was inhibited in the presence of polyvalent cations but not monovalent cations. Ca and Sr were bound preferentially over all other cations tested. Sr-binding activity was localized on the cell envelope and was sensitive to various chemical and physical pretreatments. Bound Sr was displaced by divalent ions or by H. Other monovalent ions were less effective. Bound Sr was also removed by various chelating agents. It was concluded that Sr binding by M. luteus is a reversible equilibrium process. Both ion exchange mediated by acidic cell surface components and intracellular uptake may be involved in this activity. PMID- 16348371 TI - Metal-Binding Characteristics of the Gamma-Glutamyl Capsular Polymer of Bacillus licheniformis ATCC 9945. AB - The metal-binding affinity of the anionic poly-gamma-d-glutamyl capsule of Bacillus licheniformis was investigated by using Na, Mg, Al, Ca, Cr, Mn, Fe, Ni, and Cu. Purified capsule was suspended in various concentrations of the chloride salts of the various metals, and after dialysis the bound metals were analyzed either by graphite furnace atomic absorption spectroscopy or by inductively coupled plasma-mass spectrometry. Exposure of purified capsule to excess concentrations of Na revealed it to contain 8.2 mumol of anionic sites per mg on the basis of Na binding. This was confirmed by titration of the capsule with HCl and NaOH. Other metal ions were then added in ionic concentrations equivalent to 25, 50, 75, 100, 200, and 400% of the available anionic sites. The binding characteristics varied with the metal being investigated. Addition of Cu, Al, Cr, or Fe induced flocculation. These metal ions showed the greatest affinity for B. licheniformis capsule in competitive-binding experiments. Flocculation was not seen with the addition of other metal ions. With the exception of Ni and Fe all capsule-metal-binding sites readily saturated. Ni had low affinity for the polymer, and its binding was increased at high metal concentrations. Fe binding resulted in the development of rust-colored ferrihydrite which itself could bind additional metal. Metal-binding characteristics of B. licheniformis capsule appear to be influenced by the chemical and physical properties of both the capsule and the metal ions. PMID- 16348372 TI - Bacterial degradation of ring-chlorinated acetophenones. AB - Two strains, Alcaligenes sp. strain ACA and Pseudomonas fluorescens ACB, isolated from acetophenone and 4'-hydroxyacetophenone enrichments, respectively, cometabolize a range of chlorinated acetophenones (CAs). A biological Baeyer Villiger reaction converts the CA to chlorophenyl acetate. This is evident only in the presence of an esterase inhibitor, since the CA is normally rapidly hydrolyzed to a chlorophenol which has the same substitution pattern as the original ketone. The oxygenase that attacks the ketone uses NADPH in the incorporation of one atom of O(2) and is strongly inhibited by phenols that bear an ortho or meta chlorine or bromine, but much less by cresols or phenol itself. A feedback phenomenon may thus account for the inability of strain ACA to grow on CAs, which also fail to induce the cells for their own metabolism. PMID- 16348373 TI - Chemotactic Responses of Marine Vibrio sp. Strain S14 (CCUG 15956) to Low Molecular-Weight Substances under Starvation and Recovery Conditions. AB - The chemotactic responses by starved cells of marine Vibrio sp. strain S14 differed from those elicited by cells that were not nutrient limited. The rate of chemotaxis at different concentrations of several attractants varied for starved and growing cells. Vibrio sp. strain S14 showed positive chemotaxis to leucine, valine, arginine, and glucose at the onset of energy and nutrient deprivation. A continued, though decreased, positive response was demonstrated fro leucine, arginine, and glucose at 10 h of starvation. Cells starved for 3 h displayed a stronger response to glucose than those starved for shorter or longer times. However, cells starved for 5 and 10 h responded more strongly to a lower concentration of glucose than did cells starved for 0 and 3 h. Starvation for 24 h elicited no measurable chemotaxis to leucine, arginine, or glucose. The motility decreased by over 95% in the cell population after 24 h of starvation, which resulted in a low sensitivity in the chemotaxis assay. A switch in the response to valine was observed by 3 h of starvation. The addition of nutrients of 22-h-starved cells elicited a temporary positive chemotactic response to leucine by 2 and 4 h of nutrient recovery, while cells at 1 and 6 h of recovery showed no response. At 2 h of recovery, the greatest response was recorded to 10 M leucine, whereas at 4 h it was to 10 M leucine. Ten to fifty percent of the 22 h-starved cell population regained their motility after 4 h of nutrient-aided recovery. It is possible that two types of chemosensory systems exist in marine bacteria. Starved and growing cells responded to different concentrations of the attractant, and growing cells displayed a saturated chemotactic system with leucine as the attractant, unlike the response during starvation. PMID- 16348374 TI - Streptomyces thermoautotrophicus sp. nov., a Thermophilic CO- and H(2)-Oxidizing Obligate Chemolithoautotroph. AB - The novel thermophilic CO- and H(2)-oxidizing bacterium UBT1 has been isolated from the covering soil of a burning charcoal pile. The isolate is gram positive and obligately chemolithoautotrophic and has been named Streptomyces thermoautotrophicus on the basis of G+C content (70.6 +/- 0.19 mol%), a phospholipid pattern of type II, MK-9(H(4)) as the major quinone, and other chemotaxonomic and morphological properties. S. thermoautotrophicus could grow with CO (t(d) = 8 h), H(2) plus CO(2) (t(d) = 6 h), car exhaust, or gas produced by the incomplete combustion of wood. Complex media or heterotrophic substrates such as sugars, organic acids, amino acids, and alcohols did not support growth. Molybdenum was required for CO-autotrophic growth. For growth with H(2), nickel was not necessary. The optimum growth temperature was 65 degrees C; no growth was observed below 40 degrees C. However, CO-grown cells were able to oxidize CO at temperatures of 10 to 70 degrees C. Temperature profiles of burning charcoal piles revealed that, up to a depth of about 10 to 25 cm, the entire covering soil provides a suitable habitat for S. thermoautotrophicus. The K(m) was 88 mul of CO liter and V(max) was 20.2 mul of CO h mg of protein. The threshold value of S. thermoautotrophicus of 0.2 mul of CO liter was similar to those of various soils. The specific CO-oxidizing activity in extracts with phenazinemethosulfate plus 2,6-dichlorophenolindophenol as electron acceptors was 246 mumol min mg of protein. In exception to other carboxydotrophic bacteria, S. thermoautotrophicus CO dehydrogenase was able to reduce low potential electron acceptors such as methyl and benzyl viologens. PMID- 16348375 TI - Production, Purification, and Properties of Extracellular Carboxyl Esterases from Bacillus subtilis NRRL 365. AB - Bacillus subtilis NRRL 365 produced high extracellular carboxyl esterase activity in submerged culture media containing wheat bran, corn steep liquor, and salts. Supplementation of this medium with glucose reduced esterase activity to 37% of that in the unsupplemented control. Esterase activity was purified by ammonium sulfate fractionation, DEAE-Sephadex A-50 ion-exchange chromatography with sodium chloride gradient elution, and preparative polyacrylamide gel electrophoresis. The resultant purified components, esterases I and II, manifested single bands following silver staining of polyacrylamide gel electrophoresis gels and had final specific activities of 80 and 520 U/mg, respectively. Molecular weights for components I and II were 36,000 and 105,000 to 110,000, respectively. Esterases I and II both had a pH optimum of 8.0, with relative activities of 10 and 85%, respectively, at pH 9.0. K(m)s with p-nitrophenylacetate were 0.91 mM for esterase I and 0.67 mM for esterase II. In general, patterns of enzyme inhibition were similar for both components. Differences were observed in the relative activities of esterases I and II towards p-nitrophenyl esters of acetate, propionate, and butyrate; Activity ratios for components I and II were 100:94:48 and 100:36:23, respectively. The purified components did not hydrolyze long-chain triglycerides and did not manifest proteolytic activity. PMID- 16348376 TI - Distribution of Hydrogenase Genes in Desulfovibrio spp. and Their Use in Identification of Species from the Oil Field Environment. AB - The distribution of genes for [Fe], [NiFe], and [NiFeSe] hydrogenases was determined for 22 Desulfovibrio species. The genes for [NiFe] hydrogenase were present in all species, whereas those for the [Fe] and [NiFeSe] hydrogenases had a more limited distribution. Sulfate-reducing bacteria from 16S rRNA groups other than the genus Desulfovibrio (R. Devereux, M. Delaney, F. Widdel, and D. A. Stahl, J. Bacteriol. 171:6689-6695, 1989) did not react with the [NiFe] hydrogenase gene probe, which could be used to identify different Desulfovibrio species in oil field samples following growth on lactate-sulfate medium. PMID- 16348377 TI - Pectin Lyase Activity in a Penicillium italicum Strain. AB - An extracellular pectin lyase (PNL) [poly-(methoxygalacturonide)lyase; EC 4.2.2.10] produced by Penicillium italicum CECT 2294 grown on a surface bran (natural medium) or in a submerged (synthetic medium) culture was investigated. Both culture filtrates showed macerating activity at low pH on cucumber, potato, and orange tissues. The physicochemical properties of the enzyme obtained from both culture methods were identical, as well as its catalytic properties, which were assayed by different methods. The molecular mass of the PNL obtained by gel filtration chromatography was 22 kDa; the isoelectric point was 8.6, as determined by chromatofocusing; and the enzyme was able to catalyze the eliminative cleavage of pectins with low (37%) and high (from 54 to 82%) degrees of esterification. The PNL produced in liquid medium showed a K(m) for pectin (degree of esterification, 70%) of 3.2 mg/ml, and the optimum pH was 6.0 to 7.0. This enzyme was stable at 50 degrees C and at pH 8.0. The ability of this PNL to macerate plant tissues in acidic environmental conditions, its stability at low pH and temperatures up to 50 degrees C (thus preventing mesophilic microbial growth), and the absence of pectinesterase make this preparation useful for the food industry. PMID- 16348378 TI - Occurrence of Nitrate Reductase and Molybdopterin in Xanthomonas maltophilia. AB - Fifteen of 23 ATCC strains and 2 of 9 clinical isolates of Xanthomonas maltophilia, all of which grew aerobically on ammonia, but not nitrate, as a sole nitrogen source, reduced nitrate to nitrite. X. maltophilia failed to grow anaerobically on complex medium with or without nitrate, so it is considered an obligate aerobe. Nitrate-reducing strains contained reduced methyl viologen nitrate reductase (MVH-NR) with specific activities ranging from 49.2 to 192 U mg of protein. Strain ATCC 17666 doubled its cell mass after 3 h of growth on nitrate broth under low aeration, possessed maximal MVH-NR activity, and converted the added nitrate to nitrite, which accumulated. Dissolved oxygen above 15% saturation greatly suppressed nitrite formation. All strains, except ATCC 14535, possessed between 0.25 and 5.05 pmol of molybdopterin mg of protein as measured by the Neurospora crassa nit-1 assay. The molybdopterin activity in the soluble fraction sedimented as a single symmetrical peak with an s(20,w) of 5.1. Studies identified MVH-NR in selected strains as a membrane-bound protein. The deoxycholate-solubilized MVH-NR sedimented as a single peak in sucrose density gradients with an s(20,w) of 8.8. The MVH-NR of X. maltophilia has the physical characteristics of a respiratory nitrate reductase and may enable cells to use nitrate as an electron sink under semiaerobic conditions. PMID- 16348379 TI - Adhesion of Macroconidia to the Plant Surface and Virulence of Nectria haematococca. AB - To study spore attachment of the cucurbit pathogen Nectria haematococca (anamorph, Fusarium solani f. sp. cucurbitae), mutants with adhesion-deficient macroconidia were isolated. The adhesion-deficient mutants were selected after treatment with N-methyl-N' -nitro-N-nitrosoguanidine followed by repeated enrichment for macroconidia which did not attach to polystyrene. Two independently derived mutants produced macroconidia with an approximately 50% reduction in attachment to polystyrene and to zucchini fruits. When macroconidia were inoculated into wounded zucchini fruits, the adhesion-deficient mutants were as virulent as the wild-type strain. However, in disease assays in which macroconidia were deposited onto the surface of unwounded zucchini, the mutants were less virulent than the wild type. Thus, adhesion of N. haematococca macroconidia to its host surface appears to be a virulence factor. PMID- 16348380 TI - Subunit Composition and Glycosidic Activities of the Cellulase Complex from Clostridium thermocellum JW20. AB - The subunit composition of the extracellular complex from Clostridium thermocellum was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Twenty-six bands, representing proteins with apparent molecular sizes ranging from 37,500 to 185,000 Da, could be detected by silver staining. Cultivation of the bacteria with the substrate Avicel, Sigma cellulose, Solka floc, or cellobiose as the carbon source had no influence on the number of detectable protein bands. By activity staining with the substrate carboxymethyl cellulose or xylan added to the SDS-polyacrylamide gels, 15 of the 26 bands exhibited endoglucanase activity and 13 showed xylanase activity. In 8 of the 26 bands, both activities could be found. As minor activities, beta-glucosidase, beta-xylosidase, beta-galactosidase, and beta-mannosidase activities could be demonstrated in the cellulase complex. Upon measuring the release of para nitrophenol (PNP) from PNP-cellobioside and determining the amount of glucose formed, the presence of exoglucanase activity was indicated. Upon glycoprotein staining of SDS-polyacrylamide gels, 14 of the 26 bands reacted positive, indicating the glycoprotein nature of the respective proteins. Four proteins (apparent molecular sizes, 58,000, 72,500, 94,000, and 110,000 Da) could be enriched from the originally bound cellulase complex by preparative SDS-PAGE. The two smaller proteins exhibited xylanase activity, whereas the 94,000-Da protein had endo- and exoglucanase activity, and the 110,000-Da protein degraded PNP pyranosides. PMID- 16348381 TI - Characterization of Peroxidase Secretion and Subcellular Organization of Phanerochaete chrysosporium INA-12 in the Presence of Various Soybean Phospholipid Fractions. AB - Stimulation of lignin peroxidase production by exogenous phospholipids depends on the composition of the phospholipid fraction prepared by using the Nattermann process. The fraction composed mainly of negatively charged phospholipids (NAT 89) was the most efficient source for exoprotein secretion by Phanerochaete chrysosporium INA-12. The results of biochemical marker assays and ultrastructural morphology determination by electron microscopy were correlated. Activities of succinate dehydrogenase, a mitochondrial marker, and cytochrome c oxidoreductase, an endoplasmic reticulum (ER) marker, were increased 1.3- and 2.2 fold, respectively, in the presence of NAT 89. Electron microscopy observations suggested that the amount of mitochondria and ER in culture containing phospholipids was increased at the optimum day of lignin peroxidase production. Therefore, phospholipids enhanced energetic metabolism of strain INA-12 and markedly modified fungus physiology. Since ER is involved in enzyme synthesis, we suggest that its increased amount in mycelium cultured with NAT 89 is directly associated with the higher production of lignin peroxidase. PMID- 16348382 TI - Interactions between Shewanella colwelliana, Oyster Larvae, and Hydrophobic Organophosphate Pesticides. AB - Shewanella colwelliana (strain D) is a periphytic estuarine bacterium that forms biofilms beneficial to oyster set. Our study examined whether these and other films concentrated two hydrophobic, organophosphate pesticides, Abate and malathion, that are detected in Chesapeake Bay oyster waters. Both biofilms and purified exopolysaccharide of S. colwelliana did not adsorb more of the Abate or malathion than could be accounted for by adsorption to control surfaces. Similar results were obtained by using Deleya marina, Hyphomonas MHS3, and autochthonous biofilms. Conversely, decapsulated S. colwelliana D cells, prepared in the laboratory, bioconcentrated Abate. Significantly, the S. colwelliana D biofilms exposed to Abate did not inhibit the settlement and metamorphosis of Crassostrea gigas larvae. PMID- 16348383 TI - Evidence for a Third Uptake Hydrogenase Phenotype among the Soybean Bradyrhizobia. AB - The existence of a hydrogen uptake host-regulated (Hup-hr) phenotype was established among the soybean bradyrhizobia. The Hup-hr phenotype is characterized by the expression of uptake hydrogenase activity in symbiosis with cowpea but not soybean. Uptake hydrogenase induction is not possible under free living cultural conditions by using techniques developed for uptake hydrogenase positive (Hup) Bradyrhizobium japonicum. Hydrogen oxidation by Hup-hr phenotype USDA 61 in cowpea symbioses was significant because hydrogen evolution from nitrogen-fixing nodules was not detected. An examination for uptake hydrogenase activity in soybean and cowpea with 123 strains diverse in origin and serology identified 16 Hup and 28 Hup-hr phenotype strains; the remainder appeared to be Hup. The Hup-hr phenotype was associated with serogroups 31, 76, and 94, while strains belonging to serogroups 6, 31, 110, 122, 123, and 38/115 were Hup. Hup strains of the 123 serogroup typed positive with USDA 129-specific antiserum. The presence of the uptake hydrogenase protein in cowpea bacteroids of Hup strains was demonstrated with immunoblot analyses by using antibodies against the 65-kDa subunit of uptake hydrogenase purified from strain SR470. However, the hydrogenase protein of Hup-hr strains was not detected. Results of Southern hybridization analyses with pHU1 showed the region of DNA with hydrogenase genes among Hup strains to be similar. Hybridization was also obtained with Hup-hr strains by using a variety of cloned DNA as probes including hydrogenase structural genes. Both hydrogenase structural genes also hybridized with the DNA of four Hup strains. PMID- 16348384 TI - Phenotypic Switching Affecting Chemotaxis, Xanthan Production, and Virulence in Xanthomonas campestris. AB - The chemotaxis towards sucrose and yeast extract of nine strains of Xanthomonas campestris representing pathovars campestris, armoraciae, translucens, vesicatoria, and pelargonii was analyzed by using swarm plates. Unexpectedly, each of these strains formed small or reduced swarms typical of nonmotile or nonchemotactic bacteria. With time, however, chemotactic cells appeared on the swarm plates as blebs of bacteria. These cells were strongly chemotactic and were concomitantly deficient in exopolysaccharide production. The switch from the wild type (exopolysaccharide producing and nonchemotactic) to the swarmer type (exopolysaccharide deficient and chemotactic) appeared irreversible ex planta in bacteriological medium. However, in radish leaves swarmer-type strains of X. campestris pv. campestris were able to revert to the wild type. Swarmer-type derivatives of two X. campestris pv. campestris wild-type isolates showed reduced virulence and growth in the host plants cauliflower and radish. However, exocellular complementation of X. campestris pv. campestris Hrp (nonpathogenic) mutant was achieved by coinoculation with a swarmer-type strain. PMID- 16348385 TI - Dynamics of Yeast Populations during Cider Fermentation in the Asturian Region of Spain. AB - The influence of different cidermaking systems and apple mixtures on the dynamics of yeast populations in cider manufactured in Asturias (northern Spain) were studied. Results obtained in an experimental pilot plant were compared with those found in Asturian cider plants by using traditional techniques. Saccharomyces cerevisiae and Kloeckera apiculata were found in all cases. PMID- 16348386 TI - Sheen screen, a miniaturized most-probable-number method for enumeration of oil degrading microorganisms. AB - Sheen Screen is a miniaturized method for enumerating oil-degrading microorganisms. The technique relies on the ability of oil-degrading microorganisms to emulsify oil when provided as a sole carbon source in 24-well tissue culture plates. Sediments that actively respire hydrocarbons have high numbers of Sheen Screen-positive microorganisms. PMID- 16348387 TI - Hydraulic permeability of immobilized bacterial cell aggregates. AB - A dense aggregate of cells was retained in a reactor by a supported porous membrane. A continuous flow of nutrient medium was maintained through the cell aggregate and membrane. The hydraulic resistance of the cell aggregate was monitored throughout experiments with either growing or chemically cross-linked cells, under conditions of varying flow rates. Digital image analysis was used to characterize the sizes, separations, and orientations of several thousand individual cells in electron micrographs of chemically cross-linked cell aggregates. Two nonlinear phenomena were observed. First, the hydraulic resistance varied in direct relation to and reversibly with flow rate. Second, in constant flow-rate experiments the hydraulic resistance increased with time at a faster rate than could be attributed to cell growth. Both of these phenomena were dependent upon and could be explained by the ability of cells to move with respect to one another, under the influences of Brownian motion and of convection. Such relative motion could allow changes in net alignment of cells in the direction of flow and in the volume fraction of cells in the aggregate. This explanation is consistent with image analysis data. The observed sensitivity of hydraulic resistance to flow rate was inconsistent with a model that assumed elastic deformation of individual cells, and no evidence of cell deformation was found in electron micrographs. PMID- 16348388 TI - Methylmercury decomposition in sediments and bacterial cultures: involvement of methanogens and sulfate reducers in oxidative demethylation. AB - Demethylation of monomethylmercury in freshwater and estuarine sediments and in bacterial cultures was investigated with CH(3)HgI. Under anaerobiosis, results with inhibitors indicated partial involvement of both sulfate reducers and methanogens, the former dominating estuarine sediments, while both were active in freshwaters. Aerobes were the most significant demethylators in estuarine sediments, but were unimportant in freshwater sediments. Products of anaerobic demethylation were mainly CO(2) as well as lesser amounts of CH(4). Acetogenic activity resulted in fixation of some CO(2) produced from CH(3)HgI into acetate. Aerobic demethylation in estuarine sediments produced only CH(4), while aerobic demethylation in freshwater sediments produced small amounts of both CH(4) and CO(2). Two species of Desulfovibrio produced only traces of CH(4) from CH(3)HgI, while a culture of a methylotrophic methanogen formed traces of CO(2) and CH(4) when grown on trimethylamine in the presence of the CH(3)HgI. These results indicate that both aerobes and anaerobes demethylate mercury in sediments, but that either group may dominate in a particular sediment type. Aerobic demethylation in the estuarine sediments appeared to proceed by the previously characterized organomercurial-lyase pathway, because methane was the sole product. However, aerobic demethylation in freshwater sediments as well as anaerobic demethylation in all sediments studied produced primarily carbon dioxide. This indicates the presence of an oxidative pathway, possibly one in which methylmercury serves as an analog of one-carbon substrates. PMID- 16348389 TI - Temperature effects on bacterial leaching of sulfide minerals in shake flask experiments. AB - The microbiological leaching of a sulfide ore sample was investigated in shake flask experiments. The ore sample contained pyrite, pyrrhotite, pentlandite, sphalerite, and chalcopyrite as the main sulfide minerals. The tests were performed at eight different temperatures in the range of 4 to 37 degrees C. The primary data were used for rate constant calculations, based on kinetic equations underlying two simplified models of leaching, i.e., a shrinking particle model and a shrinking core model. The rate constants thus derived were further used for the calculation of activation energy values for some of the sulfide minerals present in the ore sample. The chalcopyrite leaching rates were strongly influenced by the interaction of temperature, pH, and redox potential. Sphalerite leaching could be explained with the shrinking particle model. The data on pyrrhotite leaching displayed good fit with the shrinking core model. Pyrite leaching was found to agree with the shrinking particle model. Activation energies calculated from the rate of constants suggested that the rate-limiting steps were different for the sulfide minerals examined; they could be attributed to a chemical or biochemical reaction rather than to diffusion control. PMID- 16348390 TI - Bioconversion of alpha-Damascone by Botrytis cinerea. AB - Bioconversion of alpha-damascone (compound 1) was studied with four strains of Botrytis cinerea in grape must (pH 3.2). As biotransformation products of compound 1, 3-oxo-alpha-damascone, cis- and trans-3-hydroxy-alpha-damascone, gamma-damascenone, 3-oxo-8, 9-dihydro-alpha-damascone, and cis- and trans-3 hydroxy-8,9-dihydro-alpha-damascone were identified. In addition, acid-catalyzed chemical transformation of compound 1 to the diastereomers of 9-hydroxy-8,9 dihydro-alpha-damascone was observed. Identifications were performed by capillary gas chromatography (HRGC) and coupled HRGC techniques, i.e., on-line HRGC-mass spectrometry and HRGC-Fourier transform infrared spectroscopy, after extractive sample preparation. PMID- 16348391 TI - Genetic Mapping in the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium. AB - A method of meiotic segregation analysis based on recombinant selection in the homothallic basidiomycete Phanerochaete chrysosporium was developed. Using this method, we were able to reveal linkage relationships and to estimate recombination frequencies between seven mutations to auxotrophy. We detected two linkage groups, the first containing four and the second three of the seven mapped mutations. PMID- 16348392 TI - Effect of conditioning, betaine, and sucrose on survival of rhizobacteria in powder formulations. AB - To assess the feasibility of simplified dry formulations of beneficial rhizobacteria with improved shelf life, strains of Pseudomonas and members of the family Enterobacteriaceae were conditioned by either aging, exposure to osmotica, or growth on media amended with sucrose or betaine. Strains thus treated were formulated in 1% methylcellulose and talc, and survival was assessed over 10 to 12 months. Survival of 24-h-old cultures of the same strains suspended in 20% xanthan gum and talc over the same interval was used as the standard of comparison. The survival of strains treated with sucrose, with or without betaine, and formulated in methylcellulose and talc was equal to or greater than that of 24-h-old cultures suspended in 20% xanthan gum and talc. Aging of strains on unamended King's medium B, growth of strains on KCl-amended media, and addition of betaine to KCl-amended King's medium B, whether or not strains were suspended in buffer or a diluent isosmotic to culture conditions, failed to improve survival of the strains tested. The practicality of specific amendments, such as sucrose or other disaccharides, for promoting survival of beneficial bacteria in simplified dry formulations was thus demonstrated. PMID- 16348393 TI - Influence of the size of indigenous rhizobial populations on establishment and symbiotic performance of introduced rhizobia on field-grown legumes. AB - Indigenous rhizobia in soil present a competition barrier to the establishment of inoculant strains, possibly leading to inoculation failure. In this study, we used the natural diversity of rhizobial species and numbers in our fields to define, in quantitative terms, the relationship between indigenous rhizobial populations and inoculation response. Eight standardized inoculation trials were conducted at five well-characterized field sites on the island of Maui, Hawaii. Soil rhizobial populations ranged from 0 to over 3.5 x 10 g of soil for the different legumes used. At each site, no less than four but as many as seven legume species were planted from among the following: soybean (Glycine max), lima bean (Phaseolus lunatus), cowpea (Vigna unguiculata), bush bean (Phaseolus vulgaris), peanut (Arachis hypogaea), Leucaena leucocephala, tinga pea (Lathyrus tingeatus), alfalfa (Medicago sativa), and clover (Trifolium repens). Each legume was (i) inoculated with an equal mixture of three effective strains of homologous rhizobia, (ii) fertilized at high rates with urea, or (iii) left uninoculated. For soybeans, a nonnodulating isoline was used in all trials as the rhizobia negative control. Inoculation increased economic yield for 22 of the 29 (76%) legume species-site combinations. While the yield increase was greater than 100 kg ha in all cases, in only 11 (38%) of the species-site combinations was the increase statistically significant (P 70 ppm for 5 weeks and the concentration of 2,4-D was reduced to <100 ppm. Coincident with the accumulation of 2,4-DCP was a >400-fold decline in the numbers of fungal propagules and a marked reduction in the rate of CO(2) evolution, whereas 2,4-D did not depress either fungal propagules or respiration of the soil microbiota. 2,4-DCP did not appear to depress the numbers of total heterotrophic, sporeforming, or chitin-utilizing bacteria. In vitro and in situ assays conducted with 2,4-DCP and fungal isolates from the soil demonstrated that 2,4-DCP was toxic to fungal propagules at concentrations below those detected in the soil. PMID- 16348409 TI - Isolation and Identification of Vesicular-Arbuscular Mycorrhiza-Stimulatory Compounds from Clover (Trifolium repens) Roots. AB - Two isoflavonoids isolated from clover roots grown under phosphate stress were characterized as formononetin (7-hydroxy,4'-methoxy isoflavone) and biochanin A (5,7-dihydroxy,4'-methoxy isoflavone). At 5 ppm, these compounds stimulated hyphal growth in vitro and root colonization of an undescribed vesicular arbuscular mycorrhiza, a Glomus sp. (INVAM-112). The permethylated products of the two compounds were inactive. These findings suggest that the isoflavonoids studied may act as signal molecules in vesicular-arbuscular mycorrhiza symbiosis. PMID- 16348410 TI - Purification and Properties of beta-1, 4-Xylanase from Aeromonas caviae W-61. AB - Aeromonas caviae W-61, which was isolated from water samples at the Faculty of Agriculture, Tohoku University, produced beta-1, 4-xylanase (1,4-beta-d-xylan xylanohydrolase; EC 3.2.1.8) extracellularly. The xylanase was purified to homogeneity by using DEAE-Sephadex A-50, CM-Sephadex C-50, and Sephadex G-100 column chromatographies. The molecular weight of the purified enzyme was estimated to be 22,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of the enzyme was 9.2. The optimal pH and temperature for the activity of the enzyme were 7.0 and 55 degrees C, respectively. The enzyme was stable at pH 7.0 at temperatures of up to 50 degrees C. As enzymatic products, various xylo-oligosaccharides such as xylobiose, xylotriose, xylotetraose, and xylopentaose were formed, and only a small amount of xylose was detected. The purified enzyme did not hydrolyze starch, cellulose, carboxymethylcellulose, or beta-1, 3-xylan. PMID- 16348411 TI - Fermentation of Inulin by Clostridium thermosuccinogenes sp. nov., a Thermophilic Anaerobic Bacterium Isolated from Various Habitats. AB - Four closely related strains of thermophilic bacteria were isolated via enrichment in batch and continuous culture with inulin as the sole source of carbon and energy by using inoculations from various sources. These new strains were isolated from beet pulp from a sugar refinery, soil around a Jerusalem artichoke, fresh cow manure, and mud from a tropical pond in a botanical garden. The cells of this novel species of strictly anaerobic, gram-positive bacteria were rod shaped and nonmotile. Growth on inulin was possible between 40 and 65 degrees C, with optimum growth at 58 degrees C. All strains were capable of fermenting a large number of sugars. Formate, acetate, ethanol, lactate, H(2), and succinate were the main organic fermentation products after growth on fructose, glucose, or inulin. Synthesis of inulinase in batch culture closely paralleled growth, and the enzyme was almost completely cell bound. Strain IC is described as the type strain of a new species, Clostridium thermosuccinogenes sp. nov., with a G+C content of 35.9 mol%. PMID- 16348412 TI - Lipid content and cryotolerance of bakers' yeast in frozen doughs. AB - The relationship between lipid content and tolerance to freezing at -50 degrees C was studied in Saccharomyces cerevisiae grown under batch or fed-batch mode and various aeration and temperature conditions. A higher free-sterol-to-phospholipid ratio as well as higher free sterol and phospholipid contents correlated with the superior cryoresistance in dough or in water of the fed-batch-grown compared with the batch-grown cells. For both growth modes, the presence of excess dissolved oxygen in the culture medium greatly improved yeast cryoresistance and trehalose content (P. Gelinas, G. Fiset, A. LeDuy, and J. Goulet, Appl. Environ. Microbiol. 26:2453-2459, 1989) without significantly changing the lipid profile. Under the batch or fed-batch modes, no correlation was found between the cryotolerance of bakers' yeast and the total cellular lipid content, the total sterol content, the phospholipid unsaturation index, the phosphate or crude protein content, or the yeast cell morphology (volume and roundness). PMID- 16348413 TI - Bacterial and Fungal Numbers in Ruminal and Cecal Contents of the Blue Duiker (Cephalophus monticola). AB - Total and cellulolytic bacterial and fungal numbers were determined in ruminal and cecal contents of 20 blue duikers (Cephalophus monticola). The animals were equally divided by sex and fed two diets, either high roughage or high concentrate. The mean concentration for total bacterial numbers in the rumen was 26.0 x 10/g of contents, with values ranging from 2 x 10/g to 93 x 10/g. Cellulolytic numbers averaged 6.0 x 10/g with a range of 1.5 x 10/g to 24.0 x 10/g. No differences related to sex or diet were found. In contrast, total bacterial numbers in the cecum differed between diets (P < 0.02), i.e., 1,046 x 10 bacteria per g for animals fed the high-forage diet compared with 166 x 10/g for those fed the high-concentrate diet. Cellulolytic bacterial counts in the cecal contents averaged 3.1 and 7.0% of the total counts for the high-forage and high-concentrate diets, respectively. Low concentrations of fungi were found in both ruminal and cecal contents of some, but not all, animals. Unexpectedly, concentrations of bacteria and fungi in the rumen and cecum were highly correlated with their total numbers (concentration multiplied by total weight of contents). PMID- 16348414 TI - Ecophysiological and trophic implications of light-stimulated amino Acid utilization in marine picoplankton. AB - By using microautoradiography, light-stimulated utilization of dissolved amino acids for natural marine phytoplankton assemblages was demonstrated. The <2-mum size (diameter) picoplankton, known to be a dominant fraction of marine primary production, revealed a widespread capability for this process. Autofluorescent (chlorophyll a-containing) picoplankton and some larger phytoplankton from diverse oceanic locations, as well as isolates of the representative cyanobacterial picoplankton Synechococcus spp. (WH7803, WH8101), showed light stimulated incorporation of amino acids at trace concentrations. Dark-mediated amino acid utilization was dominated by nonfluorescent bacterial populations. Among autofluorescent picoplankton, light-stimulated exceeded dark-mediated amino acid incorporation by 5 to 75%; light-stimulated amino acid incorporation was only partially blocked by the photosystem II inhibitor 3(3,4-dichloro-phenyl)-1,1 dimethy-lurea (2 x 10 M), suggesting a photoheterotrophic incorporation mechanism. Parallel light versus dark incubations with glucose and mannitol indicated a lack of light-stimulated utilization of these nonnitrogenous compounds. Since marine primary production is frequently nitrogen limited, light mediated auxotrophic utilization of amino acids and possibly other dissolved organic nitrogen (DON) constituents may represent exploitation of the relatively large DON pool in the face of dissolved inorganic nitrogen depletion. This process (i) increases the efficiency of DON retention at the base of oceanic food webs and (ii) may in part be responsible for relatively high rates of picoplankton production under conditions of chronic dissolved inorganic nitrogen limitation. Picoplanktonic recycling of organic matter via this process has important ramifications with respect to trophic transfer via the "microbial loop." PMID- 16348415 TI - Homologous Streptomycin Resistance Gene Present among Diverse Gram-Negative Bacteria in New York State Apple Orchards. AB - The streptomycin resistance gene of Pseudomonas syringae pv. papulans Psp36 was cloned into Escherichia coli and used to develop a 500-bp DNA probe that is specific for streptomycin resistance in P. syringae pv. papulans. The probe is a portion of a 1-kb region shared by three different DNA clones of the resistance gene. In Southern hybridizations, the probe hybridized only with DNA isolated from streptomycin-resistant strains of P. syringae pv. papulans and not with the DNA of streptomycin-sensitive strains. Transposon insertions within the region of DNA shared by the three clones resulted in loss of resistance to streptomycin. Colony hybridization of bacteria isolated from apple leaves and orchard soil indicated that 39% of 398 streptomycin-resistant bacteria contained DNA that hybridized to the probe. These included all strains of P. syringae pv. papulans and some other fluorescent pseudomonads and nonfluorescent gram-negative bacteria, but none of the gram-positive bacteria. The same-size restriction fragments hybridized to the probe in P. syringae pv. papulans. Restriction fragment length polymorphism of this region was occasionally observed in strains of other taxonomic groups of bacteria. In bacteria other than P. syringae pv. papulans, the streptomycin resistance probe hybridized to different-sized plasmids and no relationship between plasmid size and taxonomic group or between plasmid size and orchard type, soil association, or leaf association could be detected. PMID- 16348417 TI - Pseudomonas stutzeri YPL-1 Genetic Transformation and Antifungal Mechanism against Fusarium solani, an Agent of Plant Root Rot. AB - An actively antagonistic bacterium that could be used as a biocontrol agent against Fusarium solani, which causes root rots with considerable losses in many important crops, was isolated from a ginseng rhizosphere and identified as a strain of Pseudomonas stutzeri. In several biochemical tests with culture filtrates of P. stutzeri YPL-1 and in mutational analyses of antifungal activities of reinforced or defective mutants, we found that the anti-F. solani mechanism of the bacterium may involve a lytic enzyme rather than a toxic substance or antibiotic. P. stutzeri YPL-1 produced extracellular chitinase and laminarinase when grown on different polymers such as chitin, laminarin, or F. solani mycelium. These lytic extracellular enzymes markedly inhibited mycelial growth rather than spore germination and also caused lysis of F. solani mycelia and germ tubes. Scanning electron microscopy revealed degradation of the F. solani mycelium. Abnormal hyphal swelling and retreating were caused by the lysing agents from P. stutzeri YPL-1, and a penetration hole was formed on the hyphae in the region of interaction with the bacterium; the walls of this region were rapidly lysed, causing leakage of protoplasm. Genetically bred P. stutzeri YPL-1 was obtained by transformation of the bacterium with a broad-host-range vector, pKT230. Also, the best conditions for the transformation were investigated. PMID- 16348416 TI - Rapid Immunocapture of Pseudomonas putida Cells from Lake Water by Using Bacterial Flagella. AB - Monoclonal antibodies to Pseudomonas putida Paw340 cells were produced. In an enzyme-linked immunosorbent assay (ELISA) against whole bacterial cells, a hybridoma cell line termed MLV1 produced a monoclonal antibody that reacted with P. putida Paw340 but showed no cross-reaction with 100 medical isolates and 150 aquatic isolates. By ELISA, immunogold electron microscopy, and Western blot (immunoblot) analysis, MLV1 antibody was found to react with purified bacterial flagella. The surfaces of magnetic polystyrene beads were coated with MLV1 antibody. By mixing MLV1 antibody-coated beads with lake water samples containing the target P. putida host, bead-cell complexes which could be recovered by attraction towards a magnet were formed. Prevention of nonspecific attachment of cells to the beads required the incorporation of detergents in the isolation protocol. These detergents affected colony-forming ability; however, the cells remained intact for direct detection. When reisolated by standard cultural methods, approximately 20% of the initial target population was recovered. Since the beads and bead-cell complexes were recovered in a magnetic field, target bacteria were separated from other lake water organisms and from particulate material which was not attracted towards the magnet and were thereby enriched. This method may now provide a useful system for recovering recombinant bacteria selectively from environmental samples. PMID- 16348418 TI - Electrophoretic and immunological comparisons of developmentally regulated proteins in members of the sclerotiniaceae and other sclerotial fungi. AB - The fungal stroma is a distinct developmental stage, a compact mass of hyphal cells enveloped by a melanized layer of rind cells which is produced from vegetative mycelium. Two types of stromata that are characteristic of members of the Sclerotiniaceae but are also produced in a wide range of other fungi, i.e., the determinate tuberlike sclerotium and the indeterminate platelike substratal stroma, were compared in these studies. Developmental proteins found in determinate sclerotial and indeterminate substratal stromata, but not in mycelia, were characterized and compared in 52 isolates of fungi, both ascomycetes (including 18 species in the Sclerotiniaceae and 5 species of Aspergillus) and the basidiomycete Sclerotium rolfsii. One-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis of mycelial, stromatal initial, and stromatal extracts demonstrated that all members of the Sclerotiniaceae produced proteins unique to stromatal extracts within a molecular weight range of 31,000 to 39,500 which composed 13 to 58% of the total protein in stromata. Proteins unique to the sclerotial stage were also produced in Sclerotium rolfsii and the Aspergillus species but within a generally lower-molecular-weight range of 11,000 to 30,000. The proteins were then characterized by two-dimensional electrophoresis to determine the number and isoelectric point of polypeptides composing each protein. Polyclonal antibodies were raised to the major 36-kDa sclerotial protein of Sclerotinia sclerotiorum (Ssp). Immunoblots demonstrated that all sclerotial proteins from species in the Sclerotiniaceae cross-reacted with anti-Ssp antibodies, while no cross-reaction was observed with proteins from substratal stromatal species in the Sclerotiniaceae, sclerotial species of Aspergillus, or Sclerotium rolfsii. Results of discriminant analysis of the data from competitive inhibition enzyme-linked immunosorbent assays were consistent with the results of immunoblotting. Three groupings, sclerotial species in the Sclerotiniaceae, substratal stromatal species in the family, and sclerotial species outside the family, were delimited on the basis of relative decreasing ability to compete for anti-Ssp antibody. These data demonstrate that stromatal proteins differ among different taxonomic groups of fungi and suggest that the Sclerotiniaceae may include two distinct lineages of genera. PMID- 16348419 TI - Rapid in situ assay for indoleacetic Acid production by bacteria immobilized on a nitrocellulose membrane. AB - We have developed a new assay that differentiates between indoleacetic acid (IAA) producing and -nonproducing bacteria on a colony plate lift. Medium supplemented with 5 mM L-tryptophan is inoculated with isolates of interest, overlaid with a nitrocellulose membrane, and then incubated until bacterial colonies reach 1 to 2 mm in diameter. The membrane is removed to a filter paper saturated with Salkowski reagent and incubated until distinct red haloes form around the colonies. The colorimetric reaction to IAA is limited to a region immediately surrounding each colony, is specific to isolates producing IAA, occurs within 1 h after the membrane is placed in the reagent, and is sensitive to as little as 50 pmol of IAA in a 2-mm spot. We have used this assay for quantifying epiphytic and endophytic populations of IAA-producing isolates of Pseudomonas syringae subsp. savastanoi and for detecting IAA-producing colonies of other pseudomonads and Erwinia herbicola. The assay provides a rapid and convenient method to screen large numbers of bacteria. PMID- 16348420 TI - Use of Polymerase Chain Reaction To Detect the Take-All Fungus, Gaeumannomyces graminis, in Infected Wheat Plants. AB - Gaeumannomyces graminis, the causative agent of take-all disease of wheat, barley, and oats, was detected in infected wheat seedlings by using the polymerase chain reaction to amplify Gaeumannomyces-specific DNA fragments. Nested primers and two rounds of amplification were used to amplify two fragments, approximately 287 and 188 bp in size, from G. graminis-infected wheat seedlings. The use of nested primers greatly decreased the number of nonspecific amplification products. Polymerase chain reaction products were not obtained with DNA from seedlings infected with several other phytopathogenic fungi or with DNA from uninfected seedlings. Amplified products were visualized on agarose gels, and their identities were confirmed by DNA hybridization. This method did not require culturing the fungus and has potential for detecting G. graminis in infested wheat, barley, or oat fields. PMID- 16348421 TI - Evolution of Volatile Sulfur Compounds during Laboratory-Scale Incubations and Indoor Preparation of Compost Used as a Substrate in Mushroom Cultivation. AB - Volatile sulfur compounds are known to be produced during the preparation of compost used as a substrate in mushroom cultivation. Because they cause odor problems, attempts have been made to reduce the production of these compounds. The influences of temperature and various additions on the production of volatile sulfur compounds from composting material were tested on laboratory-scale preparations. The production of H(2)S, COS, CH(3)SH, and (CH(3))(2)S was proven to be a biological process with an optimal temperature that coincides with the optimal temperature for biological activity. The formation of CS(2) and (CH(3))(2)S(2) was shown to be a nonbiological process. The emission of volatile sulfur compounds during the indoor preparation of mushroom compost appeared to be remarkably reduced (about 90%) as compared with the emission during the conventional outdoor process. Introduction of this indoor composting process would result in a significant reduction in environmental pollution. PMID- 16348422 TI - Thermoanaerobacter ethanolicus Growth and Product Yield from Elevated Levels of Xylose or Glucose in Continuous Cultures. AB - The performance of Thermoanaerobacter ethanolicus was evaluated in continuous culture with media containing concentrations of xylose (8 to 20 g/liter) greater than those previously reported. The ethanol yield declined from to 0.42 to 0.29 g of ethanol per g of xylose consumed when input xylose was increased from 4 to 20 g/liter. Yields of both total C(2) and C(3) products from consumed xylose and of cell biomass from ATP produced declined as the input xylose concentration was increased, which was not the case when glucose was the substrate. This suggested that yeast extract functioned as a significant energy and carbon source for cells in fermentations of xylose but not of glucose. The feasibility of this interpretation was confirmed by (i) the calculation of the products theoretically obtainable from yeast extract and (ii) the observation of significant quantities of fermentation products in inoculated sugar-free media. Markedly different patterns of metabolism for the two sugar substrates were also evidenced by the cell yield for glucose being twice that of xylose at elevated sugar concentrations. It was noted that caution must be exerted when results obtained at low xylose concentrations are extrapolated to predict those which can be obtained at higher concentrations. PMID- 16348423 TI - Hydrogen Gas Production by an Ectothiorhodospira vacuolata Strain. AB - A hydrogen gas (H(2))-producing strain of Ectothiorhodospira vacuolata isolated from Soap Lake, Washington, possessed nitrogenase activity. Increasing evolution of H(2) with decreasing ammonium chloride concentrations provided evidence that nitrogenase was the catalyst in gas production. Cells were grown in a mineral medium plus 0.2% acetate with sodium sulfide as an electron donor. Factors increasing H(2) production included addition of reduced carbon compounds such as propionate and succinate, increased reducing power by increasing sodium sulfide concentrations, and increased energy charge (ATP) by increasing light intensity. PMID- 16348424 TI - Controlling Wine Malolactic Fermentation with Nisin and Nisin-Resistant Strains of Leuconostoc oenos. AB - The polypeptide nisin (100 U/ml) prevented malolactic fermentation in wines by indigenous or intentionally added lactic acid bacteria. Nisin (100 U/ml) resistant mutants of Leuconostoc oenos were obtained and used with nisin in wine to carry out a pure-culture malolactic fermentation in the presence or absence of other lactic acid bacteria. Nisin degradation by mutants was not observed, and residual nisin was detectable in wines 4 months after it was added. Results indicated that nisin or nisin with resistant bacterial starter cultures can be used to control malolactic fermentation in wines. PMID- 16348425 TI - High-molecular-weight amylase activities from bacteria degrading starch-plastic films. AB - Amylases having unusually high molecular weights (M(r), >150,000) were found in culture supernatants of an environmentally derived microbial mixed culture selected for its ability to utilize starch-containing plastic films as sole carbon sources. The mixed culture produced amylases active at pHs 5.5 and 8.0. PMID- 16348426 TI - Temperature-Dependent Regulation by Molybdenum and Vanadium of Expression of the Structural Genes Encoding Three Nitrogenases in Azotobacter vinelandii. AB - Temperature affects the expression of the three different nitrogenases in Azotobacter vinelandii. Molybdenum repressed the vnfH and anfH operons relatively more at 30 degrees C than at 20 degrees C; at 14 degrees C molybdenum did not repress these genes at all. Similarly, V repressed the anf operon at 30 degrees C but not at 20 or 14 degrees C. Mo was poorly transported into cells grown at the lower temperatures. A. vinelandii thus has the potential to synthesize any of the three nitrogenases at 14 to 20 degrees C regardless of the presence of Mo or V. PMID- 16348427 TI - Continuous Production of Long-Side-Chain Poly-beta-Hydroxyalkanoates by Pseudomonas oleovorans. AB - Shake flask experiments showed that Pseudomonas oleovorans began to be growth inhibited at 4.65 g of sodium octanoate liter, with total inhibition at 6 g liter. In chemostat studies with 2 g of ammonium sulfate and 8 g of octanoate liter in the feed, the maximum specific growth rate was 0.51 h, and the maximum specific rate of poly-beta-hydroxyalkanoate (PHA) production was 0.074 g of PHA g of cellular protein h at a dilution rate (D) of 0.25 h. When the specific growth rate (mu) was <0.3 h, the PHA composition was relatively constant with a C(4)/C(6)/C(8)/C(10) ratio of 0.1:1.7:20.7:1.0. At mu > 0.3 h, a decrease in the percentage of C(8) with a concomitant increase in C(10) monomers as mu increased was probably due to the effects of higher concentrations of unmetabolized octanoate in the fermentor. At D = 0.24 h and an increasing carbon/nitrogen ratio, the percentage of PHA in the biomass was constant at 13% (wt/wt), indicating that nitrogen limitation did not affect PHA accumulation. Under carbon limited conditions, the yield of biomass from substrate was 0.76 g of biomass g of octanoate consumed, the yield of PHA was 0.085 g of PHA g of octanoate used, and 7.9 g of octanoate was consumed for each gram of NH(4) supplied. The maintenance coefficient was 0.046 g of octanoate g of biomass h. Replacement of sodium octanoate with octanoic acid appeared to result in transport-limited growth due to the water insolubility of the acid. PMID- 16348428 TI - Carbon Source Control of Cellobiohydrolase I and II Formation by Trichoderma reesei. AB - Regulation of the formation and secretion of two cellulase components from Trichoderma reesei QM 9414, cellobiohydrolases I and II (CBH I and CBH II, respectively), by the carbon source was investigated. With monoclonal antibodies against CBH I and CBH II it was found that during cultivation on carbon sources which enable fast growth (glucose, glycerol, and fructose), no formation of CBH I occurred, whereas low levels of CBH II were formed. Lactose and cellulose, which allow comparably slower growth, promoted the formation of both CBH I and CBH II. However, noncarbohydrate carbon sources as citrate or acetate, which also enable only slow growth, did not promote the formation of CBH I or CBH II. The addition of glucose or glycerol to lactose- or cellulose-pregrown mycelia, on the other hand, only partially reduced the formation of CBH I. This reduction was also achieved by several other metabolizable and nonmetabolizable carbon compounds, e.g., fructose, galactose, beta-methylglucoside, 2-deoxyglucose, and rhamnose, as well as by transfer to no carbon source at all. This result indicates that the control of CBH I synthesis by the carbon source is due to induction and not to repression. The use of cycloheximide and 5-fluorouracil as inhibitors at and before translation, respectively, revealed a half-life for CBH I mRNA of at least several hours, which may, at least in part, account for the prolonged synthesis of some CBH I under these conditions. Northern (RNA) hybridization with full copies of cbh1 and cbh2 genes indicated that the control of CBH I and CBH II biosyntheses by the carbon source operates mainly at the pretranslational level. We conclude that the low rate of cellulase synthesis on glucose and some other carbon sources is due to the lack of an inducer and not to carbon source repression. PMID- 16348429 TI - Identification of a Carboxylic Acid Metabolite from the Catabolism of Fluoranthene by a Mycobacterium sp. AB - A Mycobacterium sp. previously isolated from oil-contaminated estuarine sediments was capable of extensively mineralizing the high-molecular-weight polycyclic aromatic hydrocarbon fluoranthene. A carboxylic acid metabolite accumulated and was isolated by thin-layer and high-pressure liquid chromatographic analyses of ethyl acetate extracts from acidified culture media. The metabolite reached a maximum concentration of approximately 0.65% after 24 h of incubation. On the basis of comparisons with authentic compound in which we used UV and fluorescence spectrophotometry and R(f) values, as well as mass spectral and proton and carbon nuclear magnetic resonance spectral analyses, the metabolite was identified as 9 fluorenone-1-carboxylic acid. This is the first report in a microbial system of a fluoranthene metabolite in which significant degradation of one of the aromatic rings has occurred. PMID- 16348430 TI - Rate of Pyrite Bioleaching by Thiobacillus ferrooxidans: Results of an Interlaboratory Comparison. AB - Ten laboratories participated in an interlaboratory comparison of determination of bioleaching rates of a pyrite reference material. A standardized procedure and a single strain of Thiobacillus ferrooxidans were used in this study. The mean rate of bioleaching of the pyrite reference material was 12.4 mg of Fe per liter per h, with a coefficient of variation (percent relative standard deviation) of 32% as determined by eight laboratories. These results show the precision among laboratories of the determination of rates of pyrite bioleaching when a standard test procedure and reference material are used. PMID- 16348431 TI - Accumulation of trehalose and sucrose in cyanobacteria exposed to matric water stress. AB - The drought-resistant cyanobacteria Phormidium autumnale, strain LPP(4), and a Chroococcidiopsis sp. accumulated trehalose, sucrose, and both trehalose and sucrose, respectively, in response to matric water stress. Accumulated sugar concentrations reached values of up to 6.2 mug of trehalose per mug of chlorophyll in P. autumnale, 6.9 mug of sucrose per mug of chlorophyll in LPP(4), and 4.1 mug of sucrose and 3.2 mug of trehalose per mug of chlorophyll in the Chroococcidiopsis sp. The same sugars were accumulated by these cyanobacteria in similar concentrations under osmotic water stress. Cyanobacteria that did not show drought resistance (Plectonema boryanum and Synechococcus strain PCC 7942) did not accumulate significant amounts of sugars when matric water stress was applied. PMID- 16348432 TI - Population Dynamics of Rhizobium leguminosarum Tn5 Mutants with Altered Cell Surface Properties Introduced into Sterile and Nonsterile Soils. AB - The influence of cell surface properties on attachment to soil particles and on population dynamics of introduced bacteria was studied in sterilized and nonsterilized loamy sand and silt loam. Rhizobium leguminosarum RBL5523 and three Tn5 mutants (RBL5762, RBL5810, and RBL5811) with altered cell surface properties were used. Cellulose fibrils were not produced by RBL5762. Both RBL5810 and RBL5811 produced 80 to 90% less soluble exopolysaccharides and RBL5811 had, in addition, an altered lipopolysaccharide composition. In sterilized soil the total number of cells as well as the number of particle-associated cells of RBL5523 and RBL5810 were, in general, higher as compared with cell numbers of RBL5762 and RBL5811. Differences between strains in percentage of particle-associated cells in sterilized soil were only found at high inoculum densities, when populations increased little. In the nonsterilized silt loam, final population sizes, as well as numbers of particle-associated cells, of the parental strain (RBL5523) were higher than those of strains with altered cell surface properties after 56 and 112 days of incubation. But in general, differences in survival among the strains were not very marked. The importance of association with soil particles or aggregates for the survival of introduced cells was affirmed by the pronounced increase of the percentage of particle-associated cells during incubation in nonsterilized as well as sterilized soil. However, no clear relation among altered cell surface properties, particle association, and survival was found. PMID- 16348433 TI - Polygalacturonase Production by Agrobacterium tumefaciens Biovar 3. AB - Agrobacterium tumefaciens biovar 3 causes both crown gall and root decay of grape. Twenty-two Agrobacterium strains representing biovars 1, 2, and 3 were analyzed for tumorigenicity, presence of a Ti plasmid, ability to cause grape seedling root decay, and pectolytic activity. All of the biovar 3 strains, regardless of their tumorigenicity or presence of a Ti plasmid, caused root decay and were pectolytic, whereas none of the biovar 1 and 2 strains had these capacities. Isoelectrically focused gels that were activity stained with differentially buffered polygalacturonate-agarose overlays revealed that all of the biovar 3 strains produced a single polygalacturonase with a pH optimum of 4.5 and pIs ranging from 4.8 to 5.2. The enzyme was largely extracellular and was produced constitutively in basal medium supplemented with a variety of carbon sources including polygalacturonic acid. Lesions on grape seedling roots inoculated with A. tumefaciens biovar 3 strain CG49 yielded polygalacturonase activity with a pI similar to that of the enzyme produced by the bacterium in culture. These observations support the hypothesis that the polygalacturonase produced by A. tumefaciens biovar 3 has a role in grape root decay. PMID- 16348434 TI - Biodegradation of degradable plastic polyethylene by phanerochaete and streptomyces species. AB - The ability of lignin-degrading microorganisms to attack degradable plastics was investigated in pure shake flask culture studies. The degradable plastic used in this study was produced commercially by using the Archer-Daniels-Midland POLYCLEAN masterbatch and contained pro-oxidant and 6% starch. The known lignin degrading bacteria Streptomyces viridosporus T7A, S. badius 252, and S. setonii 75Vi2 and fungus Phanerochaete chrysosporium were used. Pro-oxidant activity was accelerated by placing a sheet of plastic into a drying oven at 70 degrees C under atmospheric pressure and air for 0, 4, 8, 12, 16, or 20 days. The effect of 2-, 4-, and 8-week longwave UV irradiation at 365 nm on plastic biodegradability was also investigated. For shake flask cultures, plastics were chemically disinfected and incubated-shaken at 125 rpm at 37 degrees C in 0.6% yeast extract medium (pH 7.1) for Streptomyces spp. and at 30 degrees C for the fungus in 3% malt extract medium (pH 4.5) for 4 weeks along with an uninoculated control for each treatment. Weight loss data were inconclusive because of cell mass accumulation. For almost every 70 degrees C heat-treated film, the Streptomyces spp. demonstrated a further reduction in percent elongation and polyethylene molecular weight average when compared with the corresponding uninoculated control. Significant (P < 0.05) reductions were demonstrated for the 4- and 8-day heat-treated films by all three bacteria. Heat-treated films incubated with P. chrysosporium consistently demonstrated higher percent elongation and molecular weight average than the corresponding uninoculated controls, but were lower than the corresponding zero controls (heat-treated films without 4-week incubation). The 2- and 4-week UV-treated films showed the greatest biodegradation by all three bacteria. Virtually no degradation by the fungus was observed. To our knowledge, this is the first report demonstrating bacterial degradation of these oxidized polyethylenes in pure culture. PMID- 16348435 TI - Use of a Plasmid DNA Probe To Monitor Populations of Bacillus pumilus Inoculant Strains in Hay. AB - We are evaluating naturally occurring isolates of Bacillus pumilus for use as microbial hay preservatives. Seven isolates of B. pumilus from hay contained a 42 kb cryptic plasmid (pMGD296). We wished to determine whether pMGD296 could be used as a molecular marker to follow populations of these isolates in hay over time. Southern blots and colony blots of 69 isolates of B. pumilus and other Bacillus spp. were probed with P-labeled pMGD296. Twenty-nine probe-positive isolates were identified; of these, 28 contained a plasmid with a restriction profile identical to that of pMGD296. One isolate from untreated hay contained a 40-kb plasmid (pMGD150) that was homologous to pMGD296 but had a different restriction fragment pattern. Regions of homology between the two plasmids were identified by Southern blotting, and a 1.9-kb HindIII-PstI fragment of pMGD296 lacking strong homology to pMGD150 was cloned in pUC18. The cloned fragment hybridized only with isolates containing pMGD296 and was used to estimate populations of these isolates in treated and untreated hay. PMID- 16348436 TI - Partial Purification and Characterization of a Bacteriocin Produced by Propionibacterium thoenii. AB - A partially purified bacteriocin produced by Propionibacterium thoenii designated propionicin PLG-1 was found to be active against closely related species and exhibited a broad spectrum of activity against other microorganisms. Propionicin PLG-1 was found to be heat labile, sensitive to several proteolytic enzymes, and stable at pH 3 to 9. Propionicin PLG-1 was isolated from solid medium, partially purified by ammonium sulfate precipitation, and purified further by gel filtration. Gel filtration experiments revealed that bacteriocin PLG-1 was present as two different protein aggregates with apparent molecular weights of more than 150,000 and approximately 10,000. Resolution of these protein aggregates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a protein common to both with an apparent molecular weight of 10,000. PMID- 16348437 TI - Production of Alkaline Protease by a New Aspergillus flavus Isolate under Solid Substrate Fermentation Conditions for Use as a Depilation Agent. AB - The production of alkaline protease by an Aspergillus flavus strain isolated in our laboratory by solid-substrate fermentation for use as a depilation agent and the influence of various factors on enzyme production are reported. The optimum conditions for maximum production were a growth temperature of 32 degrees C, 63% substrate moisture, and a growth period of 48 h. Enrichment with corn steep liquor or Casitone increased productivity. Scaling-up experiments indicated that flask-scale results could be reproduced at 1 and 30 kg of substrate. The enzyme preparation exhibited maximum activity at both pH 7.5 and pH 9.5. The use of this enzyme as a depilation agent was confirmed by experiments in a tannery. PMID- 16348438 TI - Methodological modifications for accurate and efficient determination of contaminant biodegradation in unsaturated calcareous soils. AB - Many techniques for quantifying microbial biodegradation of C-labeled compounds use soil-water slurries and trap mineralization-derived CO(2) in solution wells suspended within the incubation flasks. These methods are not satisfactory for studies of arid-region soils that are highly calcareous and unsaturated because (i) slurries do not simulate unsaturated conditions and (ii) the amount of CO(2) released from calcareous soils exceeds the capacity of the suspended well. This report describes simple, inexpensive methodological modifications for quantifying microbial degradation of [C]benzene and 1,2-dichloro[U-C]ethane in calcareous soils under unsaturated conditions. Soils at 50% water holding capacity were incubated with labeled contaminants for periods up to 10 weeks, followed by acidification of the soil and trapping of the evolved CO(2) in a separate container of 2 N NaOH. The CO(2) was transferred from the incubation flask to the trap solution by a gas transfer shunt containing activated charcoal to remove any volatilized labeled organics. The amount of CO(2) in the trap solution was measured by scintillation counting (disintegrations per minute). The method was tested by using two regional unamended surface soils, a sandy aridisol and a clay rich riparian soil. The results demonstrated that both [C]benzene and 1,2 dichloro[U-C]ethane were mineralized to release substantial amounts of CO(2) within 10 weeks. Levels of mineralization varied with contaminant type, soil type, and aeration status (anaerobic vs. aerobic); no significant degradation was observed in abiotic control samples. Methodological refinements of this technique resulted in total CO(2) recovery efficiency of approximately 90%. PMID- 16348439 TI - Use of ultrafiltration to isolate viruses from seawater which are pathogens of marine phytoplankton. AB - Viruses may be major structuring elements of phytoplankton communities and hence important regulators of nutrient and energy fluxes in aquatic environments. In order to ascertain whether viruses are potentially important in dictating phytoplankton community structure, it is essential to determine the extent to which representative phytoplankton taxa are susceptible to viral infection. We used a spiral ultrafiltration cartridge (30,000-molecular-weight cutoff) to concentrate viruses from seawater at efficiencies approaching 100%. Natural virus communities were concentrated from stations in the Gulf of Mexico, a barrier island pass, and a hypersaline lagoon (Laguna Madre) and added to cultures of potential phytoplankton hosts. By following changes in in vivo fluorescence over time, it was possible to isolate several viruses that were pathogens to a variety of marine phytoplankton, including a prasinophyte (Micromonas pusilla), a pennate diatom (likely a Navicula sp.), a centric diatom (of unknown taxa), and a chroococcoid cyanobacterium (a Synechococcus sp.). As well, we observed changes in fluorescence in cultures of a cryptophyte (a Rhodomonas sp.) and a chlorophyte (Nannochloropsis oculata) which were consistent with the presence of viral pathogens. Although pathogens were isolated from all stations, all the pathogens were not isolated from every station. Filterability studies on the viruses infecting M. pusilla and the Navicula sp. showed that the viruses were consistently infective after filtration through polycarbonate and glass-fiber filters but were affected by most other filter types. Establishment of phytoplankton-pathogen systems will be important in elucidating the effect that viruses have on primary producers in aquatic systems. PMID- 16348440 TI - Immunological cross-reactivities of adenosine-5'-phosphosulfate reductases from sulfate-reducing and sulfide-oxidizing bacteria. AB - Crude extracts from 14 species of sulfate-reducing bacteria comprising the genera Desulfovibrio, Desulfotomaculum, Desulfobulbus, and Desulfosarcina and from three species of sulfide-oxidizing bacteria were tested in an enzyme-linked immunosorbent assay with polyclonal antisera to adenosine 5'-phosphosulfate reductase from Desulfovibrio desulfuricans G100A. The results showed that extracts from Desulfovibrio species were all highly cross-reactive, whereas extracts from the other sulfate-reducing genera showed significantly less cross reaction. An exception was Desulfotomaculum orientis, which responded more like Desulfovibrio species than the other Desulfotomaculum strains tested. Extracts from colorless or photosynthetic sulfur bacteria were either unreactive or exhibited very low levels of reactivity with the antibodies to the enzyme from sulfate reducers. These results were confirmed by using partially purified enzymes from sulfate reducers and the most cross-reactive sulfide oxidizer, Thiobacillus denitrificans. Two types of monoclonal antibodies to adenosine 5' phosphosulfate reductase were also isolated. One type reacted more variably with the enzymes of the sulfate reducers and poorly with the Thiobacillus enzyme, whereas the second reacted strongly with Desulfovibrio, Desulfotomaculum orientis, and Thiobacillus enzymes. PMID- 16348442 TI - Leaching with Penicillium simplicissimum: Influence of Metals and Buffers on Proton Extrusion and Citric Acid Production. AB - In the presence of insoluble metal oxides (industrial filter dust, zinc oxide, synthetic mixture of metal oxides), Penicillium simplicissimum developed the ability to excrete considerable amounts of citric acid (>100 mM). Parallel with the increase of citric acid concentration in the culture broth, zinc was solubilized from zinc oxide. The adsorption of filter dust onto the mycelium (the pellets formed were less than 1 mm in diameter) was required for not only the citric acid excretion but also the leaching of zinc. When the filter dust was replaced with a synthetic mixture of metal oxides or with zinc oxide in combination with trace elements, levels of adsorption and citric acid production were observed to be similar to those in experiments where industrial filter dust was used. The two most important properties of the filter dust were its heavy metal content and its buffering capacity. These properties were simulated by adding heavy metals in soluble form (as chlorides, sulfates, or nitrates) or soluble buffers to the medium. Both heavy metals and buffers were not able to induce a citric acid efflux. As with citric acid production by Aspergillus niger, the addition of manganese lowered citric acid excretion (by 40% with metal oxide induced citric acid efflux and by 100% with urea-induced citric acid efflux). Copper antagonized the effect of manganese. The mechanism for the bulk of citric acid excretion by P. simplicissimum, however, seemed to be different from that described for citric acid accumulation by A. niger. Because of the inefficiency of metals in solubilized form and of soluble buffers to induce a strong citric acid efflux, adsorption of an insoluble metal compound (zinc oxide) turned out to be essential. Surface phenomena possibly involving the plasma membrane H-ATPase are thought to participate in the induction of citric acid excretion by P. simplicissimum in the presence of industrial filter dust. PMID- 16348441 TI - Rapid method for direct extraction of mRNA from seeded soils. AB - A protocol for direct extraction of mRNA from soil samples was developed. Soil samples (10 g) were washed twice with 120 mM phosphate buffer (pH 5.2). The lysis of cells, fixation of RNA, and hydrolysis of DNA were achieved by vigorously shaking the washed soil in a 4 M guanidine thiocyanate solution containing 25 mM sodium citrate, 0.5% sarcosyl, and 0.1 M 2-mercaptoethanol. The pH of the homogenized mixture was adjusted with 2 M sodium acetate (pH 4.0); the mRNA was then extracted with phenol and chloroform. Total RNA was precipitated with isopropanol. This method extracts up to 17 mug of total RNA per g (wet weight) of soil containing 8.0 x 10 cells of Pseudomonas aeruginosa PU21, and mRNA has been detected in 160-ng total RNA fractions. This method has been used for the detection of mRNA transcribed from specific biodegradative genes, including the nah and mer operons, in contaminated soils. This extraction method can be completed within a few hours and has tremendous potential for ecological studies of in situ gene expression among soil microbiotas. PMID- 16348443 TI - Impact of water column acidification on protozoan bacterivory at the lake sediment-water interface. AB - Although the impact of acidification on planktonic grazer food webs has been extensively studied, little is known about microbial food webs either in the water column or in the sediments. Protozoon-bacterium interactions were investigated in a chronically acidified (acid mine drainage) portion of a lake in Virginia. We determined the distribution, abundance, apparent specific grazing rate, and growth rate of protozoa over a pH range of 3.6 to 6.5. Protozoan abundance was lower at the most acidified site, while abundance, in general, was high compared with other systems. Specific grazing rates were uncorrelated with pH and ranged between 0.02 and 0.23 h, values similar to those in unacidified systems. The protozoan community from an acidified station was not better adapted (P = 0.95) to low-pH conditions than a community from an unacidified site (multivariate analysis of variance on growth rates for each community incubated at pHs 4, 5, and 6). Both communities had significantly lower (P < 0.05) growth rates at pHs 4 and 5 than at pH 6. Reduced protozoan growth rates coupled with high grazing rates and relatively higher bacterial yields (ratio of bacterial protozoan standing stock) at low pH indicate reduced net protozoan growth efficiency and a metabolic cost of acidification to the protozoan community. However, the presence of an abundant, neutrophilic protozoan community and high bacterial grazing rates indicates that acidification of Lake Anna has not inhibited the bacterium-protozoon link of the sediment microbial food web. PMID- 16348444 TI - Laboratory Study of Chemical Speciation of Mercury in Lake Sediment and Water under Aerobic and Anaerobic Conditions. AB - Chemical speciation and partitioning of radiolabeled HgCl(2) were studied in model aquatic systems consisting of undisturbed eutrophic lake sediment and water in plastic cylinders. The cylinders were either gradually made anaerobic by a gentle flow of N(2)-CO(2) or kept aerobic by air flow. The proportion of methylated Hg was significantly higher, in both water and sediment, in the anaerobic systems than in the aerobic systems. The composition and total concentration of fatty acids originating from bacterial phospholipids, as well as the concentration of vitamin B(12), including related cobalamins, were similar in sediments from the anaerobic and aerobic systems. Bacterial cell numbers were, on average, 3.6 times higher in the anaerobic water columns than in the aerobic ones. Volatilization of Hg occurred in all systems except in an autoclaved control and was of similar magnitudes in the anaerobic and aerobic systems. Incorporation of Hg into the sediment was significantly faster in the aerobic systems than in the anaerobic systems. These results suggest that episodes of anoxia in bottom waters and sediment cause an increase in net mercury methylation and, hence, an increase in bioavailable mercury. PMID- 16348445 TI - Isolation and characterization of a subsurface bacterium capable of growth on toluene, naphthalene, and other aromatic compounds. AB - A bacterium, designated F199, utilized toluene, naphthalene, dibenzothiophene, salicylate, benzoate, p-cresol, and all isomers of xylene as a sole carbon and energy source. This bacterium was isolated from Middendorf sediments, a Cretaceous age formation that underlies the Southeast Coastal Plain in South Carolina, at a depth of approximately 410 m. F199 is a gram-positive, irregular shaped bacterium that has a varied cell morphology that is dependent on culture medium type and growth stage. F199 required microaerobic conditions (40 to 80 muM O(2)) for growth on hydrocarbons, glucose, acetate, and lactate in mineral salts medium but not for growth on rich media. [C]naphthalene mineralization by F199 was induced by either naphthalene or toulene; however, [C]toluene mineralization by this strain was induced by toluene but not naphthalene. F199 was also found to harbor two plasmids larger than 100 kb. Restricted F199 plasmid and genomic DNA did not hybridize with toluene (pWW0) or naphthalene (NAH7) catabolic plasmid DNA probes. The presence in the Middendorf formation of bacteria with the capacity for degrading a variety of aromatic compounds suggests that indigenous microorganisms may have potential for in situ degradation of organic contaminants. PMID- 16348446 TI - Pathway for Biodegradation of p-Nitrophenol in a Moraxella sp. AB - A Moraxella strain grew on p-nitrophenol with stoichiometric release of nitrite. During induction of the enzymes for growth on p-nitrophenol, traces of hydroquinone accumulated in the medium. In the presence of 2,2'-dipyridyl, p nitrophenol was converted stoichiometrically to hydroquinone. Particulate enzymes catalyzed the conversion of p-nitrophenol to hydroquinone in the presence of NADPH and oxygen. Soluble enzymes catalyzed the conversion of hydroquinone to gamma-hydroxymuconic semialdehyde, which was identified by high-performance liquid chromatography (HPLC)-mass spectroscopy. Upon addition of catalytic amounts of NAD, gamma-hydroxymuconic semialdehyde was converted to beta ketoadipic acid. In the presence of pyruvate and lactic dehydrogenase, substrate amounts of NAD were required and gamma-hydroxymuconic semialdehyde was converted to maleylacetic acid, which was identified by HPLC-mass spectroscopy. Similar results were obtained when the reaction was carried out in the presence of potassium ferricyanide. Extracts prepared from p-nitrophenol-growth cells also contained an enzyme that catalyzed the oxidation of 1,2,4-benzenetriol to maleylacetic acid. The enzyme responsible for the oxidation of 1,2,4-benzenetriol was separated from the enzyme responsible for hydroquinone oxidation by DEAE cellulose chromatography. The results indicate that the pathway for biodegradation of p-nitrophenol involves the initial removal of the nitro group as nitrite and formation of hydroquinone. 1,4-Benzoquinone, a likely intermediate in the initial reaction, was not detected. Hydroquinone is converted to beta ketoadipic acid via gamma-hydroxymuconic semialdehyde and maleylacetic acid. PMID- 16348447 TI - Influence of substituents on reductive dehalogenation of 3-chlorobenzoate analogs. AB - The biochemical effects of aryl substituents on the reductive dechlorination of 3 chlorobenzoate analogs were quantified with (i) a stable 3-chlorobenzoate-grown methanogenic sludge enrichment, (ii) Desulfomonile tiedjei DCB-1, isolated from this enrichment and able to catalyze the reductive dechlorination of 3 chlorobenzoate, and (iii) a defined 3-chlorobenzoate-degrading methanogenic consortium with D. tiedjei as the key dechlorinating organism. The addition of hydrogen stimulated the dechlorination rate in the consortium. The extent of this stimulation depended on the substituent. The data were evaluated with various sets of substituent constants compiled for the Hammett equation. None of the sets yielded a satisfactory correlation between experimental values and theoretical constants. This suggests that the microbially catalyzed reductive dechlorination of 3-chlorobenzoate cannot be described simply as either a nucleophilic or an electrophilic substitution reaction. Nevertheless, observations that the presence of a para-amino or -hydroxy group inhibited the rate of dechlorination suggest that the rate-limiting step in the reductive dechlorination of 3-chlorobenzoate is a nucleophilic attack on the negatively charged pi electron cloud around the benzene nucleus. PMID- 16348448 TI - Chemotactic Behavior of Azotobacter vinelandii. AB - Chemotaxis was exhibited by Azotobacter vinelandii motile cells. Exposure of cells to sudden increases in attractant concentration suppressed the frequency of tumbling and resulted in smooth swimming. Cells responded chemotactically to a chemical gradient produced during metabolism. Motility occurred over a temperature range of 25 to 37 degrees C with an optimum pH range of between pH 7.0 and 8.0. The average speed of motile cells was determined to be 74 mum/s or 37 body lengths per s. The speed of cells appeared to increase as a function of attractant concentration. Chemotactic systems for fructose, glucose, xylitol, and mannitol were inducible. A. vinelandii exhibited chemotaxis for a number of compounds, including hexoses, hexitols, pentitols, pentoses, disaccharides, and amino sugars. We conclude from these studies that A. vinelandii exhibits a temporal chemotactic sensing system. PMID- 16348449 TI - Portable device for preparation and delivery of gas mixtures. AB - A simple, portable device for the preparation and delivery of gas mixtures has been designed and constructed. The basic feature of the device is the use of gas flow controllers to maintain stable flow rates over a wide range of downstream pressures, instead of the capillary tubes and water-filled barostats commonly used in gas-mixing devices. Elimination of the barostat avoids problems such as water leakage, the loss of gases through the barostat, and changes in gas pressure due to evaporative loss of water from the barostat. The absence of a barostat also provides a closed system, allowing the use of the device for mixing and delivering of toxic gases. The prototype of the device has been used to prepare mixtures of different gases for more than 1 year and has been found to operate consistently and reproducibly. The actual concentrations of O(2), CO(2), and N(2) in gas mixtures (determined by gas chromatography) immediately after mixing were between 2.2 and 6.6% of the desired values in four performance tests. Fluctuations in concentration of gases in mixtures after 9 days of continuous gas delivery was less than 2% in four performance tests. PMID- 16348450 TI - Pathways and microbiology of thiosulfate transformations and sulfate reduction in a marine sediment (kattegat, denmark). AB - Reductive and oxidative pathways of the sulfur cycle were studied in a marine sediment by parallel radiotracer experiments with SO(4), H(2)S, and S(2)O(3) injected into undisturbed sediment cores. The distributions of viable populations of sulfate- and thiosulfate-reducing bacteria and of thiosulfate disproportionating bacteria were concurrently determined. Sulfate reduction occurred both in the reducing sediment layers and in oxidized and even oxic surface layers. The population density of sulfate-reducing bacteria was >10 cm in the oxic layer, high enough that it could possibly account for the measured rates of sulfate reduction. The bacterial numbers counted in the reducing sediment layers were 100-fold lower. The dominant sulfate reducers growing on acetate or H(2) were gas-vacuolated motile rods which were previously undescribed. The products of sulfide oxidation, which took place in both oxidized and reduced sediment layers, were 65 to 85% S(2)O(3) and 35 to 15% SO(4). Thiosulfate was concurrently oxidized to sulfate, reduced to sulfide, and disproportionated to sulfate and sulfide. There was a gradual shift from predominance of oxidation toward predominance of reduction with depth in the sediment. Disproportionation was the most important pathway overall. Thiosulfate disproportionation occurred only as cometabolism in the marine acetate-utilizing sulfate-reducing bacteria, which could not conserve energy for growth from this process alone. Oxidative and reductive cycling of sulfur thus occurred in all sediment layers with an intermediate "thiosulfate shunt" as an important mechanism regulating the electron flow. PMID- 16348451 TI - Electron Transport in the Dissimilatory Iron Reducer, GS-15. AB - Mechanisms for electron transport to Fe(III) were investigated in GS-15, a novel anaerobic microorganism which can obtain energy for growth by coupling the complete oxidation of organic acids or aromatic compounds to the reduction of Fe(III) to Fe(II). The results indicate that Fe(III) reduction proceeds through a type b cytochrome and a membrane-bound Fe(III) reductase which is distinct from the nitrate reductase. PMID- 16348452 TI - Aerial Nodules in Casuarina cunninghamiana. AB - A complete survey of La Reunion Island showed that, in 40- to 50-year-old Casuarina cunninghamiana plantations located in the northeast at an altitude above 400 m, some trees bore aerial nodules as high as 6 to 7 m up the trunk. The nodules exhibited a significant specific acetylene reduction by the ARA method (0.77 mumol of C(2)H(4) per h/g [dry weight] of nodule) at the time of sampling (June 1990). Aerial nodules were also found on a Casuarina glauca trunk. Preliminary observations show that anatomically aerial and underground nodules do not differ significantly. In addition to host plant genetic determinants, aerial nodule formation is assumed to require sufficient rainfall, an abundance of Frankia spp. in the soil and air, and rhytidome on the tree trunk. PMID- 16348453 TI - Analysis of free amino acids in microbially colonized sandstone by precolumn phenyl isothiocyanate derivatization and high-performance liquid chromatography. AB - A fast, sensitive method for extraction and analysis of soluble free amino acids from microbially colonized sandstone is described. After precolumn phenyl isothiocyanate (PITC) derivatization, the PITC-amino acids were identified and quantified by reversed-phase high-performance liquid chromatography. This kind of analysis could be used to elucidate the role and function of amino acids in the nutrition of epi- and endolithic microorganisms active in biological weathering processes. PMID- 16348454 TI - Symbiotic Potential, Competitiveness, and Serological Properties of Bradyrhizobium japonicum Indigenous to Korean Soils. AB - The symbiotic potential of Bradyrhizobium japonicum isolates indigenous to seven Korean soils was evaluated by inoculating soybeans with 10- and 1,000-fold diluted soil suspensions (whole-soil inocula). At both levels, significant differences in the symbiotic potential of the indigenous B. japonicum isolates were demonstrated. The relationship between rhizobial numbers in the whole-soil inocula (x) and nitrogen fixation parameters (y) was best predicted by a straight line (y = a + bx) when the numbers in the inocula were 100 to 10,000 ml, while the power curve (y = ax) predicted the variation when the numbers were 1 to 100 ml. Thirty isolates from three soils showed wide differences in effectiveness (measured as milligrams of shoot N per plant), and several were of equal or greater effectiveness than reference strain B. japonicum USDA 110 on soybean cultivars Clark and Jangbaekkong. On both of the soybean cultivars grown in a Hawaiian mollisol, the Korean B. japonicum isolate YCK 213 and USDA 110 were of equal effectiveness; USDA 110 was the superior strain in colonization (nodule occupancy). Korean isolates YCK 117 and YCK 141 were superior colonizers compared with USDA 110. However, B. japonicum USDA 123 was the superior colonizer compared with isolates YCK 213, YCK 141, and YCK 117. In an immunoblot analysis of 97 indigenous Korean isolates of B. japonicum, 41% fell into the USDA 110 and USDA 123 serogroups. Serogroups USDA 110 and USDA 123 were represented in six of the seven soils examined. In one Korean soil, 100% of the B. japonicum isolates reacted only with antisera of YCK 117, an isolate from the same soil. PMID- 16348455 TI - Kinetics of Iron Oxidation by Thiobacillus ferrooxidans. AB - A statistical relationship between the rate of ferric ion production by a strain of Thiobacillus ferrooxidans and various levels of cell concentration, Fe concentration, Na concentration, and temperature was studied by a direct colorimetric method at 304 nm. The relationship was linear (90 to 93%), cross product (3 to 4%), and quadratic (1 to 2%). The levels of cell concentration and Fe concentration and their respective interactions with one another and the other factors had the most significant effects on the regression models. The solution of the quadratic response surface for optimum oxidation was a saddle point, and the predicted critical levels of temperature, cell concentration, Fe concentration, and Na concentration ranged between -6 and 2 degrees C, 0.43 and 0.62 mg/ml, 72 and 233 mM, and 29.6 mM, respectively. PMID- 16348456 TI - Immunocytochemical Identification and Localization of Active and Inactive alpha Amylase and Pullulanase in Cells of Clostridium thermosulfurogenes EM1. AB - Clostridium thermosulfurogenes EM1 formed blebs, i.e., protrusions still in contact with the cytoplasmic membrane, that originated from the cytoplasmic membrane during growth in batch culture and continuous culture. They could be observed squeezed between the cell wall and cytoplasmic membrane in cells with seemingly intact wall layers (surface layer and peptidoglycan layer) as well as in cells with wall layers in different states of degradation caused by phosphate limitation or high dilution rates. Blebs were found to turn into membrane vesicles by constriction in cases when the cell wall was heavily degraded. Bleb and vesicle formation was also observed in the absence of substrates that induce alpha-amylase and pullulanase synthesis. No correlations existed between bleb formation and the presence of active enzyme. Similar blebs could also be observed in a number of other gram-positive bacteria not producing these enzymes, but they were not observed in gram-negative bacteria. For immunoelectron-microscopic localization of alpha-amylase and pullulanase in C. thermosulfurogenes EM1, two different antisera were applied. One was raised against the enzymes isolated from the culture fluid; the other was produced against a peptide synthesized, as a defined epitope, in analogy to the N-terminal amino acid sequence (21 amino acids) of the native extracellular alpha-amylase. By using these antisera, alpha amylase and pullulanase were localized at the cell periphery in samples taken from continuous culture or batch culture. In samples prepared for electron microscopy by freeze substitution followed by ultrathin sectioning, blebs could be seen, and the immunolabel pinpointing alpha-amylase enzyme particles was seen not only randomly distributed in the cell periphery, but also lining the surface of the cytoplasmic membrane and the blebs. Cells exhibiting high or virtually no enzyme activity were labeled similarly with both antisera. This finding strongly suggests that alpha-amylase and pullulanase may occur in both active and inactive forms, depending on growth conditions. PMID- 16348457 TI - DNA-DNA Solution Hybridization Studies of the Bacterial Symbionts of Hydrothermal Vent Tube Worms (Riftia pachyptila and Tevnia jerichonana). AB - The giant tube worm, Riftia pachyptila (phylum Vestimentifera), is known only from four widely separated sulfide-rich deep-sea hydrothermal vent systems. This invertebrate is nourished by intracellular, chemoautotrophic bacterial symbionts which reside in a specialized trophosome tissue. The symbiont has not been cultured independently and is believed to be acquired de novo by host larvae of each generation. In the current study, R. pachyptila symbiont DNA was purified from the two most distant sites on the basis of its difference in density versus host DNA. These two standards were hybridized against trophosome DNAs of 13 individuals from the Guaymas Basin, Galapagos Rift, and 13 degrees N vents. This indicated that all R. pachyptila symbionts are conspecific and that the variability in DNA-DNA hybridization (relative binding ratio [RBR]) was comparable within or between widely separated vents. The symbiont of another tube worm, Tevnia jerichonana, was found to be the same as that of R. pachyptila, the first case in which distinct hosts possess the same sulfur bacterial symbiont. By contrast, Lamellibrachia sp. (same class as T. jerichonana) showed insignificant RBR with the R. pachyptila symbiont. DNA derived from solely eucaryotic tissue of R. pachyptila showed a surprisingly high RBR (20 to 50) with density-separated DNA standards. With DNAs obtained from physically separated symbionts, independent solution hybridization experiments confirmed the above-described conclusions. Possible explanations for this host-symbiont homology are discussed. PMID- 16348458 TI - Antisense mRNA-Mediated Bacteriophage Resistance in Lactococcus lactis subsp. lactis. AB - Resistance to a broad class of isometric bacteriophages that infect strains of Lactococcus lactis has been engineered into a dairy starter by expression of antisense mRNA targeted against a conserved bacteriophage gene. Maximum protection is obtained only when the entire 1,654-bp coding sequence for a 51-kDa protein is positioned in the antisense orientation with respect to a promoter sequence that functions in L. lactis subsp. lactis. Expression of the antisense mRNA results in more than 99% reduction of the total number of PFU. Plaques that do form are characterized by their relatively small size and irregular shape. A variety of truncated genes, including the open reading frame expressed in the sense orientation, fail to provide any significant measure of resistance as compared with that of the intact open reading frame. Southern hybridization with probes specific for the conserved region reveal that the [ill] plasmid constructs are maintained despite the presence of a large complement of other indigenous plasmids. Strains harboring the antisense mRNA plasmid construct grow and produce acid at a rate equivalent to that of the host strain alone, suggesting that antisense expression is not deleterious to normal cellular metabolism. PMID- 16348459 TI - Microbial formation of manganese oxides. AB - Microbial manganese oxidation was demonstrated at high Mn concentrations (5 g/liter) in bacterial cultures in the presence of a microalga. The structure of the oxide produced varied depending on the bacterial strain and mode of culture. A nonaxenic, acid-tolerant microalga, a Chlamydomonas sp., was found to mediate formation of manganite (gamma-MnOOH). Bacteria isolated from associations with crude cultures of this alga grown in aerated bioreactors formed disordered gamma MnO(2) from Mn at concentrations of 5 g/liter over 1 month, yielding 3.3 g of a semipure oxide per liter. All algal-bacterial cultures removed Mn from solution, but only those with the highest removal rates formed an insoluble oxide. While the alga was an essential component of the reaction, a Pseudomonas sp. was found to be primarily responsible for the formation of a manganese precipitate. Medium components-algal biomass and urea-showed optima at 5.7 and 10 g/liters, respectively. The scaled-up culture (50 times) gave a yield of 22.3 g (53 mg/liter/day from a 15-liter culture) of semipure disordered gamma-MnO(2), identified by X-ray diffraction and Fourier transform infrared (FTIR) spectroscopy, and had a manganese oxide O/Mn ratio of 1.92. The Mn(IV) content in the oxide was low (30.5%) compared with that of mined or chemically formed gamma MnO(2) (ca. 50%). The shortfall in the bacterial oxide manganese content was due to biological and inorganic contaminants. FTIR spectroscopy, transmission electron microscopy, and electron diffraction studies have identified manganite as a likely intermediate product in the formation of disordered gamma-MnO(2). PMID- 16348460 TI - Adhesion of Lactobacillus amylovorus to Insoluble and Derivatized Cornstarch Granules. AB - Approximately 70% of the cells in a suspension of the amylolytic bacterium Lactobacillus amylovorus bind to cornstarch granules within 30 min at 25 degrees C. More than 60% of the bound bacteria were removed by formaldehyde (2%) or glycine (1 M) at pH 2.0. More than 90% of the bound bacteria were removed by MgCl(2) (2 M; pH 7.0). Binding of L. amylovorus to cornstarch was inhibited in heat-killed cells and in cells that had been pretreated with glutaraldehyde, formaldehyde, sodium azide, trypsin, or 1% soluble potato starch. Bacterial binding to cornstarch appeared to correlate with both the concentration of cornstarch in the suspension and the amylose content in the granules. The ability of L. amylovorus to adhere to cornstarch granules was reduced for granules that had been extracted with HCl-ethanol, HCl-methanol, HCl-propanol, or HCl-butanol. Chemical derivatization of cornstarch resulted in a wide variety of adhesion responses by these bacteria. For example, 2-O-butyl starch (degree of substitution, 0.09) enhanced adhesion, whereas two palmitate starches (degree of substitution, 0.48 and 0.09) exhibited reduced adhesion activities. 2-O-(2 hydroxybutyl) starch and starch-poly(ethylene-co-acrylic acid) ester showed adhesion activities similar to those of the nonderivatized starch controls. PMID- 16348461 TI - Metabolic status of bacteria and fungi in the rhizosphere of ponderosa pine seedlings. AB - We determined the quantity and metabolic status of bacteria and fungi in rhizosphere and nonrhizosphere soil from microcosms containing ponderosa pine seedlings. Rhizosphere soil was sampled adjacent to coarse, fine, or young roots. The biovolume and metabolic status of bacterial and fungal cells was determined microscopically and converted to total and active biomass values. Cells were considered active if they possessed the ability to reduce the artificial electron acceptor 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloride (INT) to visible intracellular deposits of INT formazan. A colorimetric assay of INT formazan production was also used to assess dehydrogenase activity. INT-active microorganisms made up 44 to 55% of the microbial biomass in the soils studied. The proportion of fungal biomass that exhibited INT-reducing activity (40 to 50%) was higher than previous estimates of the active proportion of soil fungi determined by using fluorescein diacetate. Comparison between soils from different root zones revealed that the highest total and INT-active fungal biomass was adjacent to fine mycorrhizal roots, whereas the highest total and active bacterial biomass was adjacent to the young growing root tips. These observations suggest that fungi are enhanced adjacent to the fine roots compared with the nonrhizosphere soil, whereas bacteria are more responsive than fungi to labile carbon inputs in the young root zone. Colorimetric dehydrogenase assays detected gross differences between bulk and rhizosphere soil activity but were unable to detect more subtle differences due to root types. Determination of total and INT-active biomass has increased our understanding of the role of spatial compartmentalization of bacteria and fungi in rhizosphere carbon flow. PMID- 16348462 TI - Characterization of Tn5-Induced Mutants of Xenorhabdus nematophilus ATCC 19061. AB - A negative-selection vector, pHX1, was constructed for use in transposon mutagenesis of Xenorhabdus nematophilus ATCC 19061. pHX1 contains the Bacillus subtilis levansucrase gene which confers sucrose sensitivity. In addition, various Tn5-containing plasmids with different replication origins were transferred by conjugation from Escherichia coli into X. nematophilus ATCC 19061, and one of these plasmids, pGS9, yields Tn5 insertion mutants of X. nematophilus ATCC 19061. By using these two delivery vehicles, more than 250 putative Tn5 insertion mutants of X. nematophilus ATCC 19061 were isolated and were then characterized. Mutants that were altered in bromothymol blue adsorption, ability to lyse sheep erythrocytes, production of antibiotics on a variety of media, and virulence for Galleria mellonella were found. PMID- 16348463 TI - High Frequency of Natural Genetic Transformation of Pseudomonas stutzeri in Soil Extract Supplemented with a Carbon/Energy and Phosphorus Source. AB - Agar medium (SME) prepared from aqueous soil extract was used to examine genetic transformation of Pseudomonas stutzeri JM302 (his-1) by homologous his DNA in a plate transformation assay. Growth studies indicated that SME was strongly limited in carbon and nitrogen sources. Transformation was observed on SME supplemented with pyruvate, phosphate, and ammonium. A 25-fold increase of the transformation frequency was obtained with nitrogen limitation when SME was supplemented with only pyruvate plus phosphate. Similar results were obtained with artificial soil extract medium prepared on the basis of the chemical analysis of the soil extract. On a standard minimal medium, transformation frequencies also increased (10- to 60-fold) when ammonium, phosphate, or pyruvate was growth limiting. Limitation of two or three nutrients did not stimulate transformation. The size of the inoculum (2 x 10 to 2 x 10 cells) was irrelevant to the enhanced transformation under nitrogen limitation on SME or standard minimal medium. We further show that P. stutzeri can use a variety of carbon and energy sources for competence development. It is concluded that genetic transformation of P. stutzeri is possible in the chemical environment of soil upon supply of nutrients and may be strongly stimulated by a growth-limiting concentration of single nutrients including sources of C, N, or P. PMID- 16348464 TI - Arachidonic Acid production by fungi. AB - After preliminary screening, Mortierella alpina and Mortierella elongata were compared with respect to arachidonic acid content. M. alpina ATCC 16266 produced 2.1 g of arachidonic acid per liter in media containing 10% glucose while the highest percentage of arachidonic acid in lipid (43.3%) was observed at a glucose concentration of 2%. Arachidonic acid content in lipids increased to 66% during storage. PMID- 16348465 TI - Purification and Characterization of d-Aminoacylase from Alcaligenes faecalis DA1. AB - A d-aminoacylase from Alcaligenes faecalis DA1 has been purified to homogeneity by a simple purification procedure with two columns, Fractogel DEAE-650 and HW 50. The specific activity of the purified enzyme was found to be 580 U/mg of protein with N-acetyl-dl-methionine as the reaction substrate. The apparent molecular weight and isoelectric point of this enzyme were determined to be 55,000 and 5.4, respectively. PMID- 16348466 TI - Effect of Copper on Methylomonas albus BG8. AB - Addition of copper to the medium for Methylomonas albus BG8 increased cell yield and methane monooxygenase activity. Intracytoplasmic membrane was formed only in cells grown with copper supplementation. Additionally, the abundances of two major membrane proteins were affected by copper in the growth medium. These findings indicate that effects of copper on the physiology of methanotrophic bacteria are not limited to those on types II and X. PMID- 16348467 TI - Influence of Growth Conditions on the Production of a Bacteriocin, Pediocin AcH, by Pediococcus acidilactici H. AB - The influence of growth parameters on the production of pediocin AcH by Pediococcus acidilactici H was studied. This strain produced large quantities of pediocin AcH in TGE broth (Trypticase [1%], glucose [1%], yeast extract [1%], Tween 80 [0.2%], Mn [0.033 mM], Mg [0.02 mM] [pH 6.5]) within 16 to 18 h at 30 to 37 degrees C (final pH, 3.6 to 3.7). Pediocin AcH production was negligible when the pH of the medium was maintained at 5.0 or above, even in the presence of high cell mass. PMID- 16348468 TI - Evaluation of Four Aerobiological Sampling Methods for the Retrieval of Aerosolized Pseudomonas syringae. AB - The Andersen six-stage impactor, the SAS (Surface Air System) impactor, the AGI 30 impinger, and gravity plates were evaluated for the retrieval of aerosol released Pseudomonas syringae. The upper limits of the impactor samplers were exceeded at a spray concentration of 10 CFU/ml, indicating that these samplers are not appropriate for monitoring high airborne concentrations. Decreased cell concentrations were retrieved with increased sampling time for the Andersen and AGI samplers, indicating that a minimum sampling time is preferable for monitoring aerosolized vegetative cells. PMID- 16348469 TI - Effects of hydrostatic pressure on growth of hyperthermophilic archaebacteria from the juan de fuca ridge. AB - Two new strains (AL1 and AL2) of hyperthermophilic, sulfur-reducing, heterotrophic archaebacteria from high-temperature (350 degrees C) vents on the Juan de Fuca Ridge were highly barotolerant at their optimal growth temperatures (90 and 100 degrees C, respectively). A trend towards barophily at pressures greater than those encountered in situ at the sea floor was demonstrated for the more extremely thermophilic strain (AL2), implying an ability to thrive in (unexplored) habitats well below accessible vent formations. PMID- 16348470 TI - Transformation of a Methylotrophic Bacterium, Methylobacterium extorquens, with a Broad-Host-Range Plasmid by Electroporation. AB - An efficient method for the transformation of the methylotrophic bacterium Methylobacterium extorquens NR-2 with a broad-host-range plasmid, pLA2917, by electroporation was examined. Transformants of M. extorquens NR-2 expressing resistance to kanamycin were obtained after electric pulse. These transformants were found to harbor a single plasmid which was electrophoretically identical and homologous to pLA2917 obtained from Escherichia coli. Several factors which determined the transformation efficiency were optimized, resulting in a transformation efficiency of up to 8 x 10 transformants per mug of plasmid DNA by 10 pulses at a field strength of 10 kV/cm and a pulse duration of 300 mus. PMID- 16348471 TI - Aerobic microbial cometabolism of benzothiophene and 3-methylbenzothiophene. AB - A culture enriched by growth on 1-methylnaphthalene was used to study the aerobic biotransformations of benzothiophene and 3-methylbenzothiophene. Neither of the sulfur heterocyclic compounds would support growth, but they were transformed by the culture growing on 1-methylnaphthalene or glucose or peptone. Cometabolism of benzothiophene yielded benzothiophene-2,3-dione, whereas that of 3 methylbenzothiophene yielded 3-methylbenzothiophene sulfoxide and the corresponding sulfone. The identities of the dione and sulfone were verified by comparison with authentic standards. The identity of the sulfoxide was surmised from gas chromatography-mass spectrometry and gas chromatography- Fourier transform infrared spectroscopy results. Oxidation preferentially occurred at carbons 2 and 3 in benzothiophene, but when carbon 3 was substituted with a methyl group, as in 3-methylbenzothiophene, the sulfur atom was oxygenated. The predominant microorganism in the enrichment culture was a Pseudomonas strain, designated BT1, which mineralized aromatic but not aliphatic hydrocarbons. This isolate cometabolized benzothiophene and 3-methylbenzothiophene. There was no evidence that it could metabolize 3-methylbenzothiophene sulfone. When 3 methylbenzothiophene was added to Prudhoe Bay crude oil, the sulfur heterocycle was oxidized to its sulfoxide and sulfone by strain BT1 as it grew on the aromatic hydrocarbons in the crude oil. Benzothiophene-2,3-dione was found to be chemically unstable when incubated with Prudhoe Bay crude oil. Thus its formation from benzothiophene in the presence of crude oil could not be determined. PMID- 16348472 TI - Lactose Uptake Driven by Galactose Efflux in Streptococcus thermophilus: Evidence for a Galactose-Lactose Antiporter. AB - Galactose-nonfermenting (Gal) Streptococcus thermophilus TS2 releases galactose into the extracellular medium when grown in medium containing excess lactose. Starved and de-energized Gal cells, however, could be loaded with galactose to levels approximately equal to the extracellular concentration (0 to 50 mM). When loaded cells were separated from the medium and resuspended in fresh broth containing 5 mM lactose, galactose efflux occurred. De-energized, galactose loaded cells, resuspended in buffer or medium, accumulated [C]lactose at a greater rate and to significantly higher intracellular concentrations than unloaded cells. Uptake of lactose by loaded cells was inhibited more than that by unloaded cells in the presence of extracellular galactose, indicating that a galactose gradient was involved in the exchange system. When de-energized, galactose-loaded cells were resuspended in carbohydrate-free medium at pH 6.7, a proton motive force (Deltap) of 86 to 90 mV was formed, whereas de-energized, nonloaded cells maintained a Deltap of about 56 mV. However, uptake of lactose by loaded cells occurred when the proton motive force was abolished by the addition of an uncoupler or in the presence of a proton-translocating ATPase inhibitor. These results support the hypothesis that galactose efflux in GalS. thermophilus is electrogenic and that the exchange reaction (lactose uptake and galactose efflux) probably occurs via an antiporter system. PMID- 16348473 TI - The Fe Chelator Proferrorosamine A Is Essential for the Siderophore-Mediated Uptake of Iron by Pseudomonas roseus fluorescens. AB - Pseudomonas roseus fluorescens produces, besides the Fe chelator proferrorosamine A, Fe -chelating compounds, called siderophores. The production of proferrorosamine A and siderophores by P. roseus fluorescens appears to be controlled in a similar way by the concentration of available iron and by the concentration of dissolved oxygen. The higher the concentration of iron available for the microorganism, the lower the production of both chelating compounds. However, the production of siderophores was much more sensitive to iron availability than was proferrorosamine A production. Proferrorosamine A and siderophores were only produced in minimal medium C if the concentration of dissolved oxygen ranged from 4.5 to 2.0 ppm. At higher or lower concentrations, none of the iron-chelating compounds were produced. Furthermore, it has been shown that proferrorosamine-negative Tn5 mutants of P. roseus fluorescens were able to form siderophores only under iron-limiting conditions when proferrorosamine A was added to the medium. Our data suggest that proferrorosamine A production is essential for siderophore synthesis by P. roseus fluorescens; the production of siderophores occurred only when proferrorosamine A was present. PMID- 16348474 TI - Identification of para-Cresol as a Growth Factor for Methanoplanus endosymbiosus. AB - Growth of the methanogenic bacterium Methanoplanus endosymbiosus is dependent on the presence of ruminal fluid. Ruminal fluid could be replaced by the eluate of a rumen-derived anaerobic digester. From the eluate of the digester, a growth stimulatory component was purified and identified as p-cresol. Authentic p-cresol supported a half-maximal growth rate of the organism at 50 nM concentration. PMID- 16348475 TI - Production, Purification, and Characterization of beta-(1-4)-Endoxylanase of Streptomyces roseiscleroticus. AB - Twelve species of Streptomyces that formerly belonged to the genus Chainia were screened for the production of xylanase and cellulase. One species, Streptomyces roseiscleroticus (Chainia rosea) NRRL B-11019, produced up to 16.2 IU of xylanase per ml in 48 h. A xylanase from S. roseiscleroticus was purified and characterized. The enzyme was a debranching beta-(1-4)-endoxylanase showing high activity on xylan but essentially no activity against acid-swollen (Walseth) cellulose. It had a very low apparent molecular weight of 5,500 by native gel filtration, but its denatured molecular weight was 22,600 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It had an isoelectric point of 9.5. The pH and temperature optima for hydrolysis of arabinoxylan were 6.5 to 7.0 and 60 degrees C, respectively, and more than 75% of the optimum enzyme activity was retained at pH 8.0. The xylanase had a K(m) of 7.9 mg/ml and an apparent V(max) of 305 mumol . min . mg of protein. The hydrolysis rate was linear for xylan concentrations of less than 4 mg/ml, but significant inhibition was observed at xylan concentrations of more than 10 mg/ml. The predominant products of arabinoxylan hydrolysis included arabinose, xylobiose, and xylotriose. PMID- 16348476 TI - Conservation of Plasmid DNA Sequences in Coronatine-Producing Pathovars of Pseudomonas syringae. AB - In Pseudomonas syringae pv. tomato PT23.2, plasmid pPT23A (101 kb) is involved in synthesis of the phytotoxin coronatine (C. L. Bender, D. K. Malvick, and R. E. Mitchell, J. Bacteriol. 171:807-812, 1989). The physical characterization of mutations that abolished coronatine production indicated that at least 30 kb of pPT23A DNA are required for toxin synthesis. In the present study, P-labeled DNA fragments from the 30-kb region of pPT23A hybridized to plasmid DNAs from several coronatine-producing pathovars of P. syringae under conditions of high stringency. These experiments indicated that this region of pPT23A was strongly conserved in large plasmids (90 to 105 kb) that reside in P. syringae pv. atropurpurea, glycinea, and morsprunorum. The functional significance of the observed homology was demonstrated in marker-exchange experiments in which Tn5 inactivated sequences from the 30-kb region of pPT23A were used to mutate coronatine synthesis genes in the three heterologous pathovars. Physical characterization of the Tn5 insertions generated by marker exchange indicated that genes controlling coronatine synthesis in P. syringae pv. atropurpurea 1304, glycinea 4180, and morsprunorum 567 and 3714 were located on the large indigenous plasmids where homology was originally detected. Therefore, coronatine biosynthesis genes are strongly conserved in the plasmid DNAs of four producing pathovars, despite their disparate origins (California, Japan, New Zealand, Great Britain, and Italy). PMID- 16348477 TI - Identification and characterization of a pseudomonas strain capable of metabolizing phenoxybenzoates. AB - 3-Phenoxybenzoate is a transient metabolite from the breakdown of a number of pyrethroid insecticides in soil. In this study, we identified and characterized a bacterium which could grow on 3-phenoxybenzoate, converting it to phenol. On the basis of morphological and biochemical features, the 3-phenoxybenzoatedegrading isolate was determined to be a Pseudomonas species, probably a strain of Pseudomonas delafieldii, now designated Pseudomonas strain ET1. Pseudomonas strain ET1 grew on 3-phenoxybenzoate with a generation time of 3 h and a specific rate of metabolism of (2.6 +/- 0.9) x 10 g of 3-phenoxybenzoate consumed cell h. The K(m) for 3-phenoxybenzoate metabolism was 1.4 +/- 0.8 muM. The metabolism of 3-phenoxybenzoate was constitutive and not subject to catabolite repression. The metabolism of a variety of substituted diaryl ether compounds was examined. 3- and 4-Phenoxybenzoates were metabolized, but 2-phenoxybenzoate was not. Phenoxy substituted benzyl aldehyde was metabolized, but phenoxy-substituted benzyl alcohol, benzene, phenol, and aniline were not. Derivatives of 3-phenoxybenzoate substituted in the 4' position with hydroxyl, methyl, or chlorine were metabolized, yielding the corresponding 4-substituted phenol. 3-(2 Hydroxyphenoxy)benzoate was not metabolized, but 3-phenoxy-4-fluorobenzoate was. These results indicate that the metabolism of the tested diaryl ether compounds was restricted to 4-phenoxybenzoate, 3-phenoxybenzyl aldehyde, and 3 phenoxybenzoate derivatives without a substitution in the 2' position. PMID- 16348478 TI - 5'-nucleotidase activity in a eutrophic lake and an oligotrophic lake. AB - Differences in enzymatic hydrolysis of dissolved organic phosphorus and subsequent phosphorus uptake were compared by using dual-labeled (gamma-P and 2 H) ATP in oligotrophic Lake Michigan and a moderately eutrophic lake in southeastern Michigan. More than 50% of the phosphate that was hydrolyzed was immediately taken up into bacterium-sized particles in the eutrophic lake and at a near-shore site in Lake Michigan. Less than 50% of the hydrolyzed phosphate was taken up into bacterium-sized particles at an offshore site in Lake Michigan. It is hypothesized that differences in size-fractionated uptake were the result of greater phosphorus utilization capacity in bacteria in habitats where loading of organic carbon is greater. Substantial isotope dilution of labeled phosphate uptake by unlabeled phosphate occurred, which implied that the phosphate was hydrolyzed extracellularly in both systems. Comparable nucleotidase activities were measured in the eutrophic lake and Lake Michigan, but the significance of the phosphate regenerated relative to particulate phosphorus pools was an order of magnitude greater in Lake Michigan. Seventy percent of the nucleotidase activity was inhibited by 100 muM phosphate in the eutrophic lake, which suggests that most hydrolysis was by phosphatase. Therefore, nucleotidase activity may be more important to phosphorus regeneration in oligotrophic habitats than phosphatase activity. PMID- 16348479 TI - Molecular Characterization of Three Small Isometric-Headed Bacteriophages Which Vary in Their Sensitivity to the Lactococcal Phage Resistance Plasmid pTR2030. AB - Lactococcus lactis LMA12-4 is a pTR2030 transconjugant that has been used as an industrial starter culture because of its resistance to phages predominant in cheese plants. Plasmid pTR2030 interferes with susceptible phages in this host strain via two mechanisms, restriction and modification (R/M) and abortive infection (Hsp). After prolonged use of LMA12-4 transconjugants in the industry, two different bacteriophages, designated nck202.phi48 (phi48) and nck202.phi50 (phi50), were isolated which could produce plaques on LMA12-4 containing pTR2030. In this study, these two phages were characterized and compared with a third phage, nck202.phi31 (phi31), which is susceptible to both the R/M and Hsp activities encoded by pTR2030. Phage phi48 was not susceptible to inhibition by Hsp, whereas phi50 was unaffected by either the R/M or Hsp mechanisms. All three were small isometric-headed phages, but small differences were noted between the phages in the structural details of the tail base plate, susceptibility to chloroform treatment, and requirements for calcium infectivity. The phage genomes were all between 29.9 and 31.9 kb in length. Phages phi31 and phi48 harbored cohesive ends, whereas the phage phi50 genome was circularly permuted, terminally redundant, and carried a putative packaging initiation site. DNA-DNA hybridization experiments conducted between the phages revealed a common region in phi48 and phi50 that may correlate with the resistance of the two phages to the Hsp-abortive infection induced by pTR2030. Phage phi50 also harbored DNA sequences that shared homology to pTR2030 in the region where R/M activities have been localized on the plasmid. Molecular characterization of the three phages localized regions within the genomes of the pTR2030-resistant phages that may be responsible for circumventing plasmid-encoded Hsp and R/M defense mechanisms in lactococci. PMID- 16348480 TI - Comparison of representative strains of infectious hematopoietic necrosis virus by serological neutralization and cross-protection assays. AB - Infectious hematopoietic necrosis virus (IHNV) is a pathogen of young salmon and trout. Viral epizootics among these fish in private and public rearing facilities have been a problem in the northwestern United States from California to Alaska, and an IHNV vaccine has been sought by the aquaculture experts. Since an IHNV vaccine must be designed to immunize against all viral serotypes, an analysis of IHNV serotypes was made. A large number of viruses from widely separated geographic locations and different fish species had already been placed in one of five electropherotypes by the migration of the virion proteins in sodium dodecyl sulfate-polyacrylamide gels. Also, there was evidence that some of these virus isolates had differences in virulence for chinook salmon, rainbow trout, or kokanee salmon. Previous serological studies with polyclonal rabbit antisera and three IHNV isolates indicated that there was only one serotype (B. B. McCain, J. L. Fryer, and K. S. Pilcher, Proc. Soc. Exp. Biol. Med. 137:1042-1046, 1971). A substantial number of new IHNV isolations have been made since that study, and thus a more extensive comparison was made of 10 different IHNV isolates representing the five electropherotypes. This report shows that the glycoprotein from a single isolate of IHNV can induce a protective immune response in vivo to the five IHNV electropherotypes. Plaque reduction neutralization assays indicated that there was only one serotype. Thus, despite the differences observed in the migration of the structural proteins for IHNV isolated from separate geographic locations and different fish species, only one neutralizing virus type was identified. PMID- 16348481 TI - Inactivation of infectious hematopoietic necrosis virus by low levels of iodine. AB - The fish rhabdovirus infectious hematopoietic necrosis virus (IHNV) was rapidly inactivated by extremely low concentrations of iodine in water. A 99.9% virus reduction was obtained in 7.5 s when virus (10 PFU/ml) and iodine (0.1 mg/liter, final concentration) were combined in distilled-deionized or hatchery water. Iodine efficacy decreased at pHs greater than 7.5 or when proteinaceous material was added to the water. Bovine serum albumin blocked iodine inactivation of the virus more effectively than did equal concentrations of fetal bovine serum or river sediment. Sodium thiosulfate effectively neutralized free iodine. Powder, iodophor, and crystalline iodine solutions inactivated IHNV equally. Iodine rapidly inactivated IHNV isolates representing each of the five electropherotypes. Under the conditions used in this study, inactivation was not affected by temperature, salinity, or water hardness. When Dworshak National Fish Hatchery water was continuously treated to provide a free iodine concentration of 0.14 mg/liter, a 7.5-s exposure to iodine was sufficient to inactivate 99.9% of the IHNV. Iodine added to water that contained IHNV prevented infection of rainbow trout (Oncorhynchus mykiss) fry. These results suggest that the waterborne route of IHNV transmission can be blocked by adding low iodine concentrations to the water supplies of hatcheries. PMID- 16348482 TI - Characterization of the Heat Shock Response in Lactococcus lactis subsp. lactis. AB - The heat shock response in Lactococcus lactis subsp. lactis was characterized with respect to synthesis of a unique set of proteins induced by thermal stress. A shift in temperature from 30 to 42 degrees C was sufficient to arrest the growth of L. lactis subsp. lactis, but growth resumed after a shift back to 30 degrees C. Heat shock at 50 degrees C reduced the viable cell population by 10; however, pretreatment of the cells at 42 degrees C made them more thermoresistant to exposure at 50 degrees C. The enhanced synthesis of approximately 13 proteins was observed in cells labeled with S upon heat shock at 42 degrees C. Of these heat shock-induced proteins, two appeared to be homologs of GroEL and DnaK, based on their molecular weights and reactivity with antiserum against the corresponding Escherichia coli proteins. Therefore, we conclude that L. lactis subsp. lactis displays a heat shock response similar to that observed in other mesophilic bacteria. PMID- 16348483 TI - Mechanisms of h(2)s production from cysteine and cystine by microorganisms isolated from soil by selective enrichment. AB - Hydrogen sulfide (H(2)S) is a major component of biogenic gaseous sulfur emissions from terrestrial environments. However, little is known concerning the pathways for H(2)S production from the likely substrates, cysteine and cystine. A mixed microbial culture obtained from cystine-enriched soils was used in assays (50 min, 37 degrees C) with 0.05 M Tris-HCl (pH 8.5), 25 mumol of l-cysteine, 25 mumol of l-cystine, and 0.04 mumol of pyridoxal 5'-phosphate. Sulfide was trapped in a center well containing zinc acetate, while pyruvate was measured by derivatization with 2,4-dinitrophenylhydrazine. Sulfide and total pyruvate production were 17.6 and 17.2 nmol mg of protein min, respectively. Dithiothreitol did not alter reaction stoichiometry or the amount of H(2)S and total pyruvate, whereas N-ethylmaleimide reduced both H(2)S and total pyruvate production equally. The amount of H(2)S produced was reduced by 96% when only l cystine was included as the substrate in the assay and by 15% with the addition of propargylglycine, a specific suicide inhibitor of cystathionine gamma-lyase. These data indicate that the substrate for the reaction was cysteine and the enzyme responsible for H(2)S and pyruvate production was cysteine desulfhydrase (EC 4.4.1.1). The enzyme had a K(m) of 1.32 mM and was inactivated by temperatures greater than 60 degrees C. Because cysteine is present in soil and cysteine desulfhydrase is an inducible enzyme, the potential for H(2)S production by this mechanism exists in terrestrial environments. The relative importance of this mechanism compared with other processes involved in H(2)S production from soil is unknown. PMID- 16348484 TI - Degradation of 1,2,4-trichloro- and 1,2,4,5-tetrachlorobenzene by pseudomonas strains. AB - Two Pseudomonas sp. strains, capable of growth on chlorinated benzenes as the sole source of carbon and energy, were isolated by selective enrichment from soil samples of an industrial waste deposit. Strain PS12 grew on monochlorobenzene, all three isomeric dichlorobenzenes, and 1,2,4-trichlorobenzene (1,2,4-TCB). Strain PS14 additionally used 1,2,4,5-tetrachlorobenzene (1,2,4,5-TeCB). During growth on these compounds both strains released stoichiometric amounts of chloride ions. The first steps of the catabolism of 1,2,4-TCB and 1,2,4,5-TeCB proceeded via dioxygenation of the aromatic nuclei and furnished 3,4,6 trichlorocatechol. The intermediary cis-3,4,6-trichloro-1,2-dihydroxycyclohexa 3,5-diene (TCB dihydrodiol) formed from 1,2,4-TCB was rearomatized by an NAD dependent dihydrodiol dehydrogenase activity, while in the case of 1,2,4,5-TeCB oxidation the catechol was obviously produced by spontaneous elimination of hydrogen chloride from the initially formed 1,3,4,6-tetrachloro-1,2 dihydroxycyclohexa-3,5-diene. Subsequent ortho cleavage was catalyzed by a type II catechol 1,2-dioxygenase producing the corresponding 2,3,5-trichloromuconate which was channeled into the tricarboxylic acid pathway via an ordinary degradation sequence, which in the present case included 2-chloro-3-oxoadipate. From the structure-related compound 2,4,5-trichloronitrobenzene the nitro group was released as nitrite, leaving the above metabolite as 3,4,6-trichlorocatechol. Enzyme activities for the oxidation of chlorobenzenes and halogenated metabolites were induced by both strains during growth on these haloaromatics and, to a considerable extent, during growth of strain PS12 on acetate. PMID- 16348485 TI - Biodegradation by an arthrobacter species of hydrocarbons partitioned into an organic solvent. AB - An Arthrobacter strain mineralized naphthalene and n-hexadecane dissolved in 2,2,4,4,6,8,8-heptamethylnonane. The extent of mineralization increased with greater volumes of solvent. Measurements under aseptic conditions of the partitioning of naphthalene into the aqueous phase from the solid phase or from heptamethylnonane showed that the rates were rapid and did not limit mineralization. The rate of mineralization of hexadecane was rapid, although partitioning of the compound into aqueous solution was not detected. The Arthrobacter sp. grown in media with or without heptamethylnonane did not excrete products that increased the aqueous solubility of naphthalene and hexadecane. Measurements of the number of cells in the aqueous phase showed that the Arthrobacter sp. attached to the heptamethylnonane-water interface, but attachment was evident even without a substrate in the heptamethylnonane. Tests with small inocula of the Arthrobacter sp. demonstrated that at least a portion of naphthalene or hexadecane dissolved in heptamethylnonane was degraded by cells attached to the solvent-water interface. The cells did not adhere in the presence of 0.1% Triton X-100. The surfactant prevented mineralization of the hexadecane initially dissolved in heptamethylnonane, but it increased the rate and extent of mineralization of naphthalene initially dissolved in heptamethylnonane. The data show that organic solvents into which hydrophobic compounds partition affect the biodegradation of those compounds and that attachment of microorganisms to the organic solvent-water interface may be important in the transformation. PMID- 16348486 TI - Method for isolation and purification of cyanobacteria. AB - A method employing nutrient saturated glass fiber filters allowed the isolation of the same numbers of cyanobacteria from freshwater as were obtained with medium solidified with agar, while providing a 2- to 15-fold reduction in the number of accompanying heterotrophic bacteria. Imipenem, a broad-spectrum beta-lactam antibiotic which inhibits peptidoglycan biosynthesis, was superior to some other beta-lactam antibiotics for reducing the numbers of heterotrophic bacterial contaminants associated with freshly isolated cyanobacteria to a level which facilitated the production of axenic cyanobacterial cultures. PMID- 16348487 TI - Production of Cell Wall-Degrading Enzymes by the Phytopathogenic Fungus Sclerotinia sclerotiorum. AB - The range of polysaccharide-degrading enzymes and glycosidases formed by the phytopathogenic fungus Sclerotinia sclerotiorum was monitored following growth on 16 carbohydrate substrates. Endo- and exoenzymes capable of degrading cellulosic, hemicellulosic, and pectinolytic polysaccharides were secreted. Pectinolytic activities were produced constitutively on all of the substrates tested. Cellulolytic enzymes were not induced in simple sugar (i.e., glucose or xylose) media. Polysaccharide growth substrates and cellulase inducers increased all of the enzyme activities tested. Gel filtration analysis revealed the appearance of new molecular forms of pectinase, beta-xylosidase, and cellobiosidase during induction on pectin and carboxymethyl cellulose media. PMID- 16348488 TI - Flavonoids released naturally from alfalfa promote development of symbiotic glomus spores in vitro. AB - Because flavonoids from legumes induce transcription of nodulation genes in symbiotic rhizobial bacteria, it is reasonable to test whether these compounds alter the development of vesicular-arbuscular mycorrhizal (VAM) fungi that infect those plants. Quercetin-3-O-galactoside, the dominant flavonoid released naturally from alfalfa (Medicago sativa L.) seeds, promoted spore germination of Glomus etunicatum and Glomus macrocarpum in vitro. Quercetin produced the maximum increases in spore germination, hyphal elongation, and hyphal branching in G. etunicatum at 1 to 2.5 muM concentrations. Two flavonoids exuded from alfalfa roots, 4',7-dihydroxyflavone and 4',7-dihydroxyflavanone, also enhanced spore germination of this fungal species. Formononetin, an isoflavone that is released from stressed alfalfa roots, inhibited germination of both Glomus species. These in vitro results suggest that plant flavonoids may facilitate or regulate the development of VAM symbioses and offer new hope for developing pure, plant-free cultures of VAM fungi. PMID- 16348489 TI - Ultrastructural changes in an obligately barophilic marine bacterium after decompression. AB - The bacterial isolate MT-41 from 10,476 m, nearly the greatest ocean depth, is obligately barophilic. The purpose of this study was to describe the morphological changes in MT-41 due to nearly isothermal decompression followed by incubation at atmospheric pressure. Two cultures were grown at 103.5 MPa and 2 degrees C and then decompressed to atmospheric pressure (0.101 MPa). One of the cultures was fixed just before decompression. The other culture, kept at 0 degrees C, was sampled immediately and four more times over 168 h. The number of CFU (assayed at 103.5 MPa and 2 degrees C) declined with incubation time at atmospheric pressure. Decompression itself did not lead to immediate morphological changes. The ultrastructure, however, was altered with increasing time at atmospheric pressure. The first aberrations were intracellular vesicles and membrane fragments in the medium. After these changes were plasmolysis, cell lysis, the formation of extracellular vesicles, and the formation of ghost cells. Intact cells in the longest incubation at atmospheric pressure had the normal cytoplasmic granularity suggestive of ribosomes but had few and poorly stained fibrils in the bacterial nucleoids. From the practical standpoint, samples of hadal deep-sea regions need to be fixed either in situ or shortly after arrival at the sea surface even when recovered in insulated sampling gear. This should prevent drastic structural degradation of sampled cells, thus allowing both accurate estimates of deep-sea benthic standing stock and realistic morphological descriptions. PMID- 16348490 TI - Conditional-suicide containment system for bacteria which mineralize aromatics. AB - A model conditional-suicide system to control genetically engineered microorganisms able to degrade substituted benzoates is reported. The system is based on two elements. One element consists of a fusion between the promoter of the Pseudomonas putida TOL plasmid-encoded meta-cleavage pathway operon (P(m)) and the lacI gene encoding Lac repressor plus xylS, coding for the positive regulator of P(m). The other element carries a fusion between the P(tac) promoter and the gef gene, which encodes a killing function. In the presence of XylS effectors, LacI protein is synthesized, preventing the expression of the killing function. In the absence of effectors, expression of the P(tac)::gef cassette is no longer prevented and a high rate of cell killing is observed. The substitution of XylS for XylSthr45, a mutant regulator with altered effector specificity and increased affinity for benzoates, allows the control of populations able to degrade a wider range of benzoates at micromolar substrate concentrations. Given the wide effector specificity of the key regulators, the wild-type and mutant XylS proteins, the system should allow the control of populations able to metabolize benzoate; methyl-, dimethyl-, chloro-, dichloro-, ethyl-, and methoxybenzoates; salicylate; and methyl- and chlorosalicylates. A small population of genetically engineered microorganisms became Gef resistant; however, the mechanism of such survival remains unknown. PMID- 16348491 TI - Gene transfer in marine water column and sediment microcosms by natural plasmid transformation. AB - We investigated the possibility for natural transformation in the marine environment by using broad-host-range plasmid multimers and a high-frequency-of transformation (HFT) Vibrio strain as the recipient. Water and sediment samples were taken from Tampa Bay, the eastern Gulf of Mexico, the Florida Shelf near Miami, and the Bahamas Bank. In water column microcosms, transformation frequencies ranged from 1.7 x 10 to 2.7 x 10 transformants per recipient, with highest frequencies occurring when low levels of nutrients (peptone and yeast extract) were added. The presence of the ambient community either reduced transformation frequency by an order of magnitude or had no effect. In sterile sediments, nutrient additions had no consistent effect on transformation, with transfer frequencies similar to those observed in the water column. Transformation was not observed in any sediment experiment when the ambient microbial community was present. These findings are the first report of natural plasmid transformation in seawater and in the presence of the ambient microbial community. This process may be a mechanism for the acquisition of small, nonconjugative plasmids, which are commonly found in aquatic bacteria. Our data also suggest that natural transformation may be more likely to occur in the water column than in native marine sediments, contradicting prior conclusions based on studies with sterile sediments. PMID- 16348492 TI - Subgroups of the Cowpea Miscellany: Symbiotic Specificity within Bradyrhizobium spp. for Vigna unguiculata, Phaseolus lunatus, Arachis hypogaea, and Macroptilium atropurpureum. AB - Rhizobia classified as Bradyrhizobium spp. comprise a highly heterogeneous group of bacteria that exhibit differential symbiotic characteristics on hosts in the cowpea miscellany cross-inoculation group. To delineate the degree of specificity exhibited by four legumes in the cowpea miscellany, we tested the symbiotic characteristics of indigenous cowpea bradyrhizobia on cowpea (Vigna unguiculata), siratro (Macroptilium atropurpureum), lima bean (Phaseolus lunatus), and peanut (Arachis hypogaea). The most-probable-number counts of indigenous bradyrhizobia at three sites on Maui, Hawaii, were substantially different on the four hosts: highest on siratro, intermediate on cowpea, and significantly lower on both lima bean and peanut. Bradyrhizobia from single cowpea nodules from the most-probable number assays were inoculated onto the four hosts. Effectiveness patterns of these rhizobia on cowpea followed a normal distribution but were strikingly different on the other legumes. The effectiveness profiles on siratro and cowpea were similar but not identical. The indigenous cowpea-derived bradyrhizobia were of only moderate effectiveness on siratro and were in all cases lower than the inoculant-quality reference strain. Between 5 and 51% of the bradyrhizobia, depending on site, failed to nodulate peanut, whereas 0 to 32% failed to nodulate lima bean. No significant correlation was observed between the relative effectiveness of the bradyrhizobia on cowpea and their corresponding effectiveness on either lima bean or peanut. At all sites, bradyrhizobia that were ineffective on cowpea but that effectively nodulated lima bean, peanut, or both were found. Eighteen percent or fewer of the bradyrhizobia were as effective on lima bean as the reference inoculant strain; 44% or fewer were as effective on peanut as the reference strain. Only 18% of all cowpea-derived bradyrhizobia tested were able to form N(2)-fixing nodules on both lima bean and peanut. These results indicate the need to measure indigenous bradyrhizobial population characteristics directly with the crop of interest to obtain an accurate assessment of the need to inoculate. PMID- 16348493 TI - Transfer and Expression of the Catabolic Plasmid pBRC60 in Wild Bacterial Recipients in a Freshwater Ecosystem. AB - 3-Chlorobenzoate (3Cba)-degrading bacteria were isolated from the waters and sediments of flowthrough mesocosms dosed with various concentrations of 3Cba and inoculated with a 3Cba-degrading Alcaligenes sp., strain BR60. Bacteria capable of 3Cba degradation which were distinct from BR60 were isolated. They carried pBRC60, a plasmid introduced with Alcaligenes sp. strain BR60 that carries a transposable element (Tn5271) encoding 3Cba degradation. The isolates expressed these genes in different ways. The majority of pBRC60 recipients were motile, yellow-pigmented, gram-negative rods related to the group III pseudomonads and to BR60 by substrate utilization pattern. They were capable of complete 3Cba degradation at both millimolar and micromolar concentrations. Two isolates, Pseudomonas fluorescens PR24B(pBRC60) and Pseudomonas sp. strain PR120(pBRC60), are more distantly related to BR60 and both produced chlorocatechol when exposed to 3Cba at millimolar concentrations in the presence of yeast extract. These species showed poor growth in liquid 3Cba minimal medium but could degrade 3Cba in continuous cultures dosed with micromolar levels of the chemical. Laboratory matings confirm that pBRC60 can transfer from BR60 to species in both the beta and gamma subgroups of the proteobacteria and that 3Cba gene expression is variable between species. Selection pressures acting on pBRC60 recipients are discussed. PMID- 16348494 TI - Pseudomonas putida Which Can Grow in the Presence of Toluene. AB - A Pseudomonas putida strain able to grow in the presence of more than 50% toluene was isolated from soil. The strain was tolerant of other toxic solvents, including aliphatic hydrocarbons, alicyclic hydrocarbons, aromatic hydrocarbons, alcohols, and ethers. The stability of the solvent tolerance of strain IH-2000 was stimulated by addition of Mg and Ca to the medium containing toluene. PMID- 16348495 TI - Biotransformation of the Pentahydroxy Flavone Quercetin by Rhizobium loti and Bradyrhizobium Strains (Lotus). AB - Lotus rhizobia catabolized quercetin in an arabinose-based medium via a novel form of C-ring cleavage, yielding phloroglucinol and protocatechuic acid. Conservation of the A and B rings of the flavone suggests that a chalcone could be formed as a transient intermediate. PMID- 16348496 TI - Mineralization of linear alkylbenzene sulfonate by a four-member aerobic bacterial consortium. AB - A bacterial consortium capable of linear alkylbenzene sulfonate (LAS) mineralization under aerobic conditions was isolated from a chemostat inoculated with activated sludge. The consortium, designated KJB, consisted of four members, all of which were gram-negative, rod-shaped bacteria that grew in pairs and short chains. Three isolates had biochemical properties characteristic of Pseudomonas spp.; the fourth showed characteristics of the Aeromonas spp. Cell suspensions were grown together in minimal medium with [C]LAS as the only carbon source. After 13 days of incubation, more than 25% of the [C]LAS was mineralized to CO(2) by the consortium. Pure bacterial cultures and combinations lacking any one member of the KJB bacterial consortium did not mineralize LAS. Three isolates carried out primary biodegradation of the surfactant, and one did not. This study shows that the four bacteria complemented each other and synergistically mineralized LAS, indicating catabolic cooperation among the four consortium members. PMID- 16348497 TI - Phenotypic Diversity among Strains of Bradyrhizobium japonicum Belonging to Serogroup 110. AB - Thirty-four strains of Bradyrhizobium japonicum within serogroup 110 were examined for phenotypic diversity. The strains differed in their abilities to nodulate and fix dinitrogen with Glycine max (L.) Merr. cv. Williams. Thirteen strains expressed uptake hydrogenase activity when induced as free-living cultures in the presence of 2% hydrogen and oxygen. Six bacteriophage susceptibility reactions were observed. Each of the strains produced either a large, mucoid or a small, dry colony morphology, but colony type was not related to effectiveness for nitrogen fixation. PMID- 16348498 TI - Reductive ortho and meta Dechlorination of a Polychlorinated Biphenyl Congener by Anaerobic Microorganisms. AB - We used gas chromatography-mass spectrometry to study the metabolic fate of 2,3,5,6-tetrachlorobiphenyl (2356-CB) (350 muM) incubated with unacclimated methanogenic pond sediment. The 2356-CB was dechlorinated to 25-CB (21%), 26-CB (63%), and 236-CB (16%) in 37 weeks. This is the first experimental demonstration of ortho dechlorination of a polychlorinated biphenyl by anaerobic microorganisms. PMID- 16348499 TI - Monohydroxylation of Phenol and 2,5-Dichlorophenol by Toluene Dioxygenase in Pseudomonas putida F1. AB - [This corrects the article on p. 2650 in vol. 55.]. PMID- 16348500 TI - Bacterial and Fungal Numbers in Ruminal and Cecal Contents of the Blue Duiker (Cephalophus monticola). AB - [This corrects the article on p. 469 in vol. 57.]. PMID- 16348501 TI - Distribution of DNA Sequences Encoding Narrow- and Broad-Spectrum Mercury Resistance. AB - The distribution of DNA sequences homologous with three mer genes was determined in unselected and mercury-resistant water and sediment isolates. The maximum proportions of unselected bacterial isolates containing DNA hybridizing with the 358merA, 358merB, and 501merR probes, derived from gram-negative organisms, were 93.8, 21, and 100%, respectively. Up to 53.3% of mercury chloride-resistant isolates and 54% of methylmercury hydroxide-resistant isolates did not contain DNA homologous with 358merA or 358merB, respectively. Hybridizations performed at high and low stringencies demonstrated that divergence of the merA gene accounted for many of the mercury-resistant but probe-negative isolates. Sixteen mercury resistant Bacillus spp. isolated from the least contaminated site all contained DNA homologous with 258merA, originally from a gram-positive organism, but only four hybridized weakly with 358merA. The results demonstrate the wide distribution of mercury resistance genes but, because of the diversity of genetic determinants, highlight the importance of using multiple detection techniques and gene probes derived from a variety of origins for such studies. PMID- 16348502 TI - Procedure for Obtaining Efficient Root Nodulation of a Pea Cultivar by a Desired Rhizobium Strain and Preempting Nodulation by Other Strains. AB - The specificity between the sym-2 gene bred into certain cultivars of pea (Pisum sativum L.) and the nodX gene, present only rarely in isolates of Rhizobium leguminosarum, can be exploited to preempt competition or nodulation blocking by a Rhizobium strain indigenous to a soil environment. The principle is to isolate an R. leguminosarum strain prevalent in a locale, convert it into a strain that will nodulate a desirable pea cultivar carrying sym-2 by establishing nodX in it, and then use the resulting Rhizobium strain with the pea cultivar carrying sym-2. To accomplish this, we first constructed a transposon Tn5 derivative called Tn5nodX and an efficient delivery vehicle that is suicidal in R. leguminosarum. We tested the potential utility of the system in greenhouse experiments. The results are encouraging enough to warrant extensive experiments under field conditions. PMID- 16348503 TI - Evidence that beta-Galactosidase of Sulfolobus solfataricus Is Only One of Several Activities of a Thermostable beta-d-Glycosidase. AB - A survey of Sulfolobus isolates showed all to contain thermostable enzyme activities hydrolyzing various glycosidic compounds. Of those not previously reported, the beta-glucosidase activity of Sulfolobus solfataricus isolate P2 was chosen for further study and found to have the same kinetics of inactivation, apparent molecular weight, and many (though not all) other biochemical properties of the beta-galactosidase also present in this strain. The two activities copurified approximately 850-fold to apparent homogeneity. The enzyme, whose subunit M(r) was estimated to be 60,000 to 65,000 by gel permeation chromatography of the active enzyme and 70,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis of the denatured form, hydrolyzed a variety of low-molecular-weight, beta-linked glycosides and could account for most of the corresponding activities found in crude extract. Kinetic analyses indicated that chromogenic beta-d-galactosides and beta-d-glucosides are hydrolyzed at a common active site and that beta-glucosides and beta-fucosides represent the preferred substrates. The liberation of aglycone from aryl beta-d-glucosides was stimulated by alcohols in a manner suggesting specific interaction between alcohol and enzyme. PMID- 16348504 TI - Specificity of Activated CryIA Proteins from Bacillus thuringiensis subsp. kurstaki HD-1 for Defoliating Forest Lepidoptera. AB - The insecticidal activity of the CryIA(a), CryIA(b), and CryIA(c) toxins from Bacillus thuringiensis subsp. kurstaki HD-1 was determined in force-feeding experiments with larvae of Choristoneura fumiferana, C. occidentalis, C. pinus, Lymantria dispar, Orgyia leucostigma, Malacosoma disstria, and Actebia fennica. The toxins were obtained from cloned protoxin genes expressed in Escherichia coli. The protoxins were activated with gut juice from Bombyx mori larvae. Biological activity of the individual gene products as well as the native HD-1 toxin was assessed as the dose which prevented 50% of the insects from producing frass within 3 days (frass failure dose [FFD(50)]). The three toxins were about equally active against M. disstria. In the Choristoneura species, CryIA(a) and CryIA(b) were up to fivefold more toxic than CryIA(c). In the lymantriid species, CryIA(a) and CryIA(b) were up to 100-fold more toxic than CryIA(c). The toxicity of HD-1 was similar to that of the individual CryIA(a) or CryIA(b) toxins in all of these species. None of the CryIA toxins or HD-1 exhibited and toxicity towards A. fennica. Comparison of the observed FFD(50) of HD-1 with the FFD(50) expected on the basis of its crystal composition suggested a possible synergistic effect of the toxins in the two lymantriid species. Our results further illustrate the diversity of activity spectra of these highly related proteins and provide a data base for studies with forest insects to elucidate the molecular basis of toxin specificity. PMID- 16348505 TI - DNA Synthesis and Tritiated Thymidine Incorporation by Heterotrophic Freshwater Bacteria in Continuous Culture. AB - Continuous cultivation of heterotrophic freshwater bacteria was used to assess the relationship between DNA synthesis and tritiated thymidine incorporation. The bacteria were grown on a yeast extract medium with generation times of 0.25 to 3.7 days. In six different continuous cultures, each inoculated with a grazer free mixed bacterial sample from Lake Vechten (The Netherlands), tritiated thymidine incorporation into a cold trichloroacetic acid precipitate and bacterial cell production were measured simultaneously. Empirical conversion factors were determined by division of both parameters. They ranged from 0.25 x 10 to 1.31 x 10 cells mol of tritiated thymidine (mean, 0.60 x 10 cells mol of tritiated thymidine). In addition, DNA concentrations were measured by fluorometry with Hoechst 33258. The validity of this technique was confirmed. Down to a generation time of 0.67 day, bacterial DNA content showed little variation, with values of 3.8 to 4.9 fg of DNA cell. Theoretical conversion factors, which can be derived from DNA content under several assumptions, were between 0.26 x 10 and 0.34 x 10 cells mol of thymidine (mean, 0.30 x 10 cells mol of thymidine). Isotope dilution was considered the main factor in the observed discrepancy between the conversion factors. In all experiments, a tritiated thymidine concentration of 20 nM was used. Control experiments indicated maximum incorporation at this concentration. It was therefore concluded that the observed difference resulted from intracellular isotope dilution which cannot be detected by current techniques for isotope dilution analysis. PMID- 16348506 TI - Chemical Characterization of Polysaccharide from the Slime Layer of the Cyanobacterium Microcystis flos-aquae C3-40. AB - Macromolecular material from the slime layer of the cyanobacterium Microcystis flos-aquae C3-40 was defined as material that adhered to cells during centrifugation in growth medium but was dislodged by washing with deionized water and retained within dialysis tubing with a molecular-weight cutoff of 3,500. At each step of this isolation procedure, the slime was observed microscopically. Cells in the centrifugal pellet were surrounded by large amounts of slime that excluded negative stain, whereas cells that had been washed with water lacked visible slime. Two independently isolated lots of slime contained no detectable protein (<1%, wt/wt) and consisted predominantly of anthrone-reacting polysaccharide. Sugars in a hydrolysate of slime polysaccharide were derivatized with trimethylsilylimidazole and examined by gas chromatography-mass spectrometry. The composition of the slime polysaccharide was 1.5% (wt/wt) galactose, 2.0% glucose, 3.0% xylose, 5.0% mannose, 5.5% rhamnose, and 83% galacturonic acid. This composition resembles that of the plant polysaccharide pectin, which was treated in parallel as a control. Consistent with earlier indications that M. flos-aquae slime preferentially binds certain cations, the ratio of Fe to Na in the dialyzed slime was 10 times that in the growth medium. The composition of the slime is discussed with respect to possible mechanisms of cation binding in comparison with other cyanobacterial exopolysaccharides and pectin. PMID- 16348507 TI - Detection of Gene Expression in Genetically Engineered Microorganisms and Natural Phytoplankton Populations in the Marine Environment by mRNA Analysis. AB - A simple method that combines guanidinium isothiocyanate RNA extraction and probing with antisense and sense RNA probes is described for analysis of microbial gene expression in planktonic populations. Probing of RNA sample extracts with sense-strand RNA probes was used as a control for nonspecific hybridization or contamination of mRNA with target DNA. This method enabled detection of expression of a plasmid-encoded neomycin phosphotransferase gene (nptII) in as few as 10Vibrio cells per ml in 100 ml of seawater. We have used this method to detect expression of the ribulose-1,5-bisphosphate carboxylase large-subunit gene (rbcL) in Synechococcus cultures and natural phytoplankton populations in the Dry Tortugas, Florida. During a 36-h diel study, rbcL expression of the indigenous phytoplankton was greatest in the day, least at night (1100, 0300, and 0100 h), and variable at dawn or dusk (0700 and 1900 h). These results are the first report of gene expression in natural populations by mRNA isolation and probing. This methodology should be useful for the study of gene expression in microorganisms released into the environment for agricultural or bioremediation purposes and indigenous populations containing highly conserved target gene sequences. PMID- 16348508 TI - Diversity and population dynamics of methanogenic bacteria in a granular consortium. AB - Upflow anaerobic sludge blanket bioreactor granules were used as an experimental model microbial consortium to study the dynamics and distribution of methanogens. Immunologic methods revealed a considerable diversity of methanogens that was greater in mesophilic granules than in the same granules 4 months after a temperature shift from 38 to 55 degrees C. During this period, the sizes of the methanogenic subpopulations changed with distinctive profiles after the initial reduction caused by the shift. Methanogens antigenically related to Methanobrevibacter smithii PS and ALI, Methanobacterium hungatei JF1, and Methanosarcina thermophila TM1 increased rapidly, reached a short plateau, and then fell to lower concentrations that persisted for the duration of the experiment. A methanogen related to Methanogenium cariaci JR1 followed a similar profile at the beginning, but it soon diminished below detection levels. Methanothrix rods weakly related to the strain Opfikon increased rapidly, reaching a high-level, long-lasting plateau. Two methanogens related to Methanobrevibacter arboriphilus AZ and Methanobacterium thermoautotrophicum DeltaH emerged from very low levels before the temperature shift and multiplied to attain their highest numbers 4 months after the shift. Histochemistry and immunohistochemistry revealed thick layers, globular clusters, and lawns of variable density which were distinctive of the methanogens related to M. thermoautotrophicum DeltaH, M. thermophila TM1, and M. arboriphilus AZ and M. soehngenii Opfikon, respectively, in thin sections of granules grown at 55 degrees C for 4 months. Mesophilic granules showed a different pattern of methanogenic subpopulations. PMID- 16348509 TI - DNA Polymorphisms in Lentinula edodes, the Shiitake Mushroom. AB - DNA restriction fragment length polymorphisms (RFLPs) were examined in Lentinula edodes strains. Genomic DNA from strain 70 was cloned in plasmid vector pUC19, and 18 random clones containing low-copy DNA sequences were used to probe seven strains in Southern DNA-DNA hybridizations. Each cloned fragment revealed DNA polymorphism. An RFLP genotype was determined for each strain and the genetic relatedness was assessed. The coefficients of genetic similarity among the seven strains ranged from 0.43 to 0.90. The inheritance of RFLP markers was examined in single spore isolates. Homokaryons displayed a loss of polymorphic bands compared with the parent dikaryon. Hybrids constructed by crossing compatible homokaryons displayed the inheritance of RFLP markers from each parent homokaryon. PMID- 16348510 TI - Control of the Lysine Biosynthesis Sequence in Corynebacterium glutamicum as Analyzed by Overexpression of the Individual Corresponding Genes. AB - The gene cluster that codes for feedback-resistant aspartate kinase (lysCalpha and lysCbeta) and aspartate semialdehyde dehydrogenase (asd) was cloned from a mutant strain of Corynebacterium glutamicum. Its functional analysis by subcloning, enzyme assays, and type of aspartate kinase regulation enabled the isolation of a fragment for separate expression of the feedback-resistant kinase without aspartate semialdehyde dehydrogenase expression. This was used together with other clones constructed (J. Cremer, L. Eggeling, and H. Sahm, Mol. Gen. Genet. 220:478-480, 1990) to overexpress individually each of the six genes that convert aspartate to lysine. Analysis of lysine formation revealed that overexpression of the feedback-resistant kinase alone suffices to achieve lysine formation (38 mM). Also, sole overexpression of wild-type dihydrodipicolinate synthase resulted in lysine formation but in a lower amount (11 mM). The other four enzymes had no effect on lysine secretion. With a plasmid overexpressing both relevant enzymes together, a further increase in lysine yield was obtained. This shows that of the six enzymes that convert aspartate to lysine the kinase and the synthase are responsible for flow control in the wild-type background and can be useful for construction of lysine-producing strains. PMID- 16348511 TI - Characterization of the Cell Wall-Bound Proteinase of Lactobacillus casei HN14. AB - Lactobacillus casei HN14, which was isolated from homemade cheese, produces an extracellular, cell wall-bound proteinase. The HN14 proteinase can be removed from the cell envelope by washing the cells in a Ca-free buffer. The activity of the crude proteinase extract is inhibited by phenylmethylsulfonyl fluoride, showing that the enzyme is a serine-type proteinase. Considering the substrate specificity, the HN14 proteinase is similar to the lactococcal PI-type enzyme, since it hydrolyzes beta-casein only. Lactobacillus casei HN14 appeared to be plasmid free, which suggests that the proteinase gene is chromosomally located. Chromosomal DNA of this strain hybridizes with DNA probes Q1 (which contains a fragment of the prtM gene) and Q6 and Q92 (which contain fragments of the prtP gene); all three probes originated from the proteinase gene region of Lactococcus lactis subsp. cremoris Wg2. A restriction enzyme map of the proteinase region of Lactobacillus casei HN14 was constructed on the basis of hybridization experiments. Comparison of the restriction enzyme maps of the Lactobacillus casei HN14 proteinase gene region and those of lactococcal proteinase gene regions studied so far indicates that they are highly similar. PMID- 16348513 TI - Genetic Variation of Lactobacillus delbrueckii subsp. lactis Bacteriophages Isolated from Cheese Processing Plants in Finland. AB - The genomes of four Lactobacillus delbrueckii subsp. lactis bacteriophages were characterized by restriction endonuclease mapping, Southern hybridization, and heteroduplex analysis. The phages were isolated from different cheese processing plants in Finland between 1950 and 1972. All four phages had a small isometric head and a long noncontractile tail. Two different types of genome (double stranded DNA) organization existed among the different phages, the pac type and the cos type, corresponding to alternative types of phage DNA packaging. Three phages belonged to the pac type, and a fourth was a cos-type phage. The pac-type phages were genetically closely related. In the genomes of the pac-type phages, three putative insertion/deletions (0.7 to 0.8 kb, 1.0 kb, and 1.5 kb) and one other region (0.9 kb) containing clustered base substitutions were discovered and localized. At the phenotype level, three main differences were observed among the pac-type phages. These concerned two minor structural proteins and the efficiency of phage DNA packaging. The genomes of the pac-type phages showed only weak homology with that of the cos-type phage. Phage-related DNA, probably a defective prophage, was located in the chromosome of the host strain sensitive to the cos type phage. This DNA exhibited homology under stringent conditions to the pac type phages. PMID- 16348512 TI - Degradation of the Herbicide Glyphosate by Members of the Family Rhizobiaceae. AB - Several strains of the family Rhizobiaceae were tested for their ability to degrade the phosphonate herbicide glyphosate (isopropylamine salt of N phosphonomethylglycine). All organisms tested (seven Rhizobium meliloti strains, Rhizobium leguminosarum, Rhizobium galega, Rhizobium trifolii, Agrobacterium rhizogenes, and Agrobacterium tumefaciens) were able to grow on glyphosate as the sole source of phosphorus in the presence of the aromatic amino acids, although growth on glyphosate was not as fast as on P(i). These results suggest that glyphosate degradation ability is widespread in the family Rhizobiaceae. Uptake and metabolism of glyphosate were studied by using R. meliloti 1021. Sarcosine was found to be the immediate breakdown product, indicating that the initial cleavage of glyphosate was at the C-P bond. Therefore, glyphosate breakdown in R. meliloti 1021 is achieved by a C-P lyase activity. PMID- 16348514 TI - Evaluation of Polymyxa betae Keskin Contaminated by Beet Necrotic Yellow Vein Virus in Soil. AB - The fungus Polymyxa betae Keskin belongs to the family Plasmodiophoraceae and lives in the soil as an obligatory parasite of the roots of the Chenopodiaceae. When contaminated by beet necrotic yellow vein virus, this viruliferous fungus causes a serious disease of sugar beet known as rhizomania, whereas the infection by the fungus alone (aviruliferous fungus) causes only slight damage to the plant with little economic consequence. The manifestation of rhizomania in sugar beet is directly related to the concentration of infecting units of viruliferous P. betae present in the soil. (One infecting unit is a group of one or more sporosori that liberate zoospores capable of visibly infecting a plant.) By using current methods of analysis, it is possible to estimate the total quantity of P. betae present in the soil, but one cannot distinguish quantitatively the infecting units of aviruliferous from viruliferous P. betae. A new method has been developed based on the technique of the most probable number and enzyme linked immunosorbent assay to estimate the concentration of infecting units of viruliferous P. betae in soil. The method is suitable for the routine analysis of numerous soil samples and allows one to estimate the concentration of viable forms of the fungus P. betae, whether or not contaminated by beet necrotic yellow vein virus, present in a soil affected by rhizomania or presumed healthy. The analyses performed with this method are economical and use a reagent kit and equipment in wide use. PMID- 16348515 TI - Incompatibility of Lactobacillus Vectors with Replicons Derived from Small Cryptic Lactobacillus Plasmids and Segregational Instability of the Introduced Vectors. AB - Three new Lactobacillus vectors based on cryptic Lactobacillus plasmids were constructed. The shuttle vector pLP3537 consists of a 2.3-kb plasmid from Lactobacillus pentosus MD353, an erythromycin resistance gene from Staphylococcus aureus plasmid pE194, and pUC19 as a replicon for Escherichia coli. The vectors pLPE317 and pLPE323, which do not contain E. coli sequences, were generated by introducing the erythromycin resistance gene of pE194 into a 1.7- and a 2.3-kb plasmid from L. pentosus MD353, respectively. These vectors and the shuttle vector pLP825 (M. Posno, R. J. Leer, J. M. M. van Rijn, B. C. Lokman, and P. H. Pouwels, p. 397-401, in A. T. Ganesan and J. A. Hoch, ed., Genetics and biotechnology of bacilli, vol. 2, 1988) could be introduced by electroporation into Lactobacillus casei, L. pentosus, L. plantarum, L. acidophilus, L. fermentum, and L. brevis strains with similar efficiencies. Transformation efficiencies were strain dependent and varied from 10 to 10 transformants per mug of DNA. Plasmid DNA analysis of L. pentosus MD353 transformants revealed that the introduction of pLP3537 or pLPE323 was invariably accompanied by loss of the endogenous 2.3-kb plasmid. Remarkably, pLPE317 could only be introduced into an L. pentosus MD353 strain that had been previously cured of its endogenous 1.7-kb plasmid. The curing phenomena are most likely to be explained by the incompatibility of the vectors and resident plasmids. Lactobacillus vectors are generally rapidly lost when cells are cultivated in the absence of selective pressure. However, pLPE323 is stable in three of four Lactobacillus strains tested so far. PMID- 16348516 TI - Bacteriocin-Producing Strain of Lactococcus lactis subsp. diacitilactis S50. AB - Lactococcus lactis subsp. diacitilactis S50 produces a bacteriocin, designated bacteriocin S50, which has a narrow antibacterial spectrum. It was active only against Lactococcus species, including a nisin producer exhibiting a bactericidal effect. The activity of bacteriocin S50 was sensitive to proteases. It retained antimicrobial activity after being heated to 100 degrees C for up to 60 min and in the pH range 2 to 11. PMID- 16348517 TI - Correlation between Possession of a Respiration-Dependent Na Pump and Na Requirement for Growth of Marine Bacteria. AB - The possession of a respiration-dependent primary sodium pump and the requirement of Na for growth were investigated in bacterial isolates from marine environments. The bacteria in which NADH oxidase specifically required Na for maximum activity were believed to possess a primary sodium pump. All bacteria that failed to grow without the addition of NaCl possessed a primary Na pump. All bacteria that had no primary Na pump grew without additional NaCl. The primary Na pump seems to be involved in the Na requirement of marine bacteria, and this can be regarded as a criterion for the definition of marine bacteria. PMID- 16348518 TI - Hydrogen Oxidation by the Host-Controlled Uptake Hydrogenase Phenotype of Bradyrhizobium japonicum in Symbiosis with Soybean Host Plants. AB - Symbioses between uptake hydrogenase host-regulated (Hup-hr) phenotypes of Bradyrhizobium japonicum and exotic, agronomically unadapted soybean germ plasm were examined for expression of uptake hydrogenase activity. Determinations for hydrogen evolution and uptake hydrogenase activity identified five plant introduction (PI) lines which formed hydrogen-oxidizing symbioses with strains USDA 61 and PA3 6c. Hup-hr strains belonging to serogroup 94 expressed uptake hydrogenase activity in symbioses with PI 181696 and PI 219655 at rates sufficient to prevent hydrogen from escaping the nodules. The identification of soybean germ plasm forming hydrogen-oxidizing symbioses with Hup-hr bradyrhizobia potentially has implications for enhancing nitrogen fixation efficiency in soybean production. PMID- 16348519 TI - Unexpected Errors in Gas Chromatographic Analysis of Methane Production by Thermophilic Bacteria. AB - Unexpected errors in methane measurement by gas chromatography occurred when samples at thermophilic temperatures were analyzed. With a standard curve prepared at room temperature (25 degrees C), stoppered bottles incubated and sampled at 37 to 85 degrees C showed more methane upon analysis than bottles incubated at 25 degrees C: values at 50, 63, and 85 degrees C were 109, 126, and 125%, respectively, of the 25 degrees C value. All variation between 4 and 50 degrees C can be explained by the temperature difference between culture bottle and sampling syringe, and the variation of methane concentration can be predicted by the gas law. Between 50 and 63 degrees C, there was a more dramatic rise than predicted by theory. These variations are important to consider if thermophilic methane production is to be measured accurately. Methods to avoid errors are discussed. PMID- 16348520 TI - Laboratory-based model of microbiologically induced corrosion of copper. AB - The interactions of bacteria isolated from corroded copper coupons on thin films of copper evaporated onto germanium internal reflection elements were evaluated nondestructively in real time by attentuated total reflectance Fourier transform infrared spectroscopy. The films were stable in the presence of flowing or static sterile culture medium. When exposed to and colonized by the bacterium CCI 8, the copper thin film corroded. Corrosion was enhanced under quiescent conditions. In conjunction with corrosion of the copper thin film was an increase in the concentration of polysaccharide material at the copper-biofilm interface. A different bacterium (CCI 11) did not corrode the copper thin film, and the establishment of this bacterium on the copper surface prevented corrosion of the thin film by CCI 8. PMID- 16348521 TI - Anaerobic Growth and Fermentation Characteristics of Paecilomyces lilacinus Isolated from Mullet Gut. AB - The anaerobic growth and fermentation of a marine isolate of Paecilomyces lilacinus is described. The fungus was isolated from mullet gut and grew optimally at 30 degrees C and at a salinity of >/=10%. The best growth was obtained with glucose or laminarin as substrate, and the growth yield was 5.0 g (dry weight of fungus) per mol of hexose fermented. Moles of products as a percentage of moles of hexose fermented were acetate, 29.0%; ethanol, 156.6%; CO(2), 108.0%; and lactate, 4.3%. Together these products accounted for >80% of hexose carbon. Hydrogen and formate were not detectable as fermentation end products (<0.5%). Other substrates utilized for growth, although less effectively than laminarin or glucose, included the monosaccharides galactose, fructose, arabinose, and xylose and the disaccharides maltose and cellobiose. No growth of the fungus occurred on cellulose, and of a variety of other polysaccharides tested only xylan supported growth. PMID- 16348522 TI - Production and degradation of oxalic Acid by brown rot fungi. AB - Our results show that all of the brown rot fungi tested produce oxalic acid in liquid as well as in semisolid cultures. Gloeophyllum trabeum, which accumulates the lowest amount of oxalic acid during decay of pine holocellulose, showed the highest polysaccharide-depolymerizing activity. Semisolid cultures inoculated with this fungus rapidly converted C-labeled oxalic acid to CO(2) during cellulose depolymerization. The other brown rot fungi also oxidized C-labeled oxalic acid, although less rapidly. In contrast, semisolid cultures inoculated with the white rot fungus Coriolus versicolor did not significantly catabolize the acid and did not depolymerize the holocellulose during decay. Semisolid cultures of G. trabeum amended with desferrioxamine, a specific iron-chelating agent, were unable to lower the degree of polymerization of cellulose or to oxidize C-labeled oxalic acid to the extent or at the rate that control cultures did. These results suggest that both iron and oxalic acid are involved in cellulose depolymerization by brown rot fungi. PMID- 16348523 TI - Construction of an Acid-Tolerant Rhizobium leguminosarum Biovar Trifolii Strain with Enhanced Capacity for Nitrogen Fixation. AB - Strain ANU1173 is an acid-tolerant Rhizobium leguminosarum biovar trifolii strain that is able to nodulate subterranean clover plants growing in agar culture at pH 4.4 At pH 6.5, its symbiotic effectiveness in association with Trifolium subterraneum cv. Mt. Barker was 80% relative to that of strain ANU794, a Sm derivative of the commercial inoculant R. leguminosarum bv. trifolii TA1. Strain ANU1173 contained four indigenous megaplasmids, the smallest of these being the symbiotic (Sym) plasmid. The critical pH requirement for growth of strain ANU1173 in laboratory media was shown not to be associated with this plasmid. When the Sym plasmid of strain ANU1173(pSym-1173) was mobilized into a Nod strain of R. leguminosarum bv. viciae, the plasmid conferred to the transconjugant a level of symbiotic effectiveness in association with T. subterraneum that was similar to that observed with ANU1173. The symbiotic effectiveness of strain ANU1173 was improved by first curing pSym-1173 (generating strain ANU1184) and replacing it with another R. leguminosarum bv. trifolii Sym plasmid, pBR1AN. Subterranean clover plants inoculated with strain ANU1184 (pBR1AN) exhibited a 35 or 53% increase in acetylene reduction activity and a 20 or 17% increase in dry weight when grown at pH 6.5 and pH 4.4, respectively, compared with plants inoculated with strain ANU1173 and grown under the same pH conditions. It was further shown that pBR1AN was stably maintained in strain ANU1184 under free-living and symbiotic conditions. These results indicate that it is possible to construct an acid-tolerant strain of R. leguminosarum bv. trifolii with an enhanced capacity for nitrogen fixation. PMID- 16348524 TI - Biochemical basis for whole-cell uptake kinetics: specific affinity, oligotrophic capacity, and the meaning of the michaelis constant. AB - Formulations are presented that describe the concentration dependency of nutrient limited transport and growth in molecular terms. They relate the rate of transport at steady state through a two-sequence process, transport and metabolism, to ambient concentrations according to the amounts and kinetic characteristics of the two rate-limiting proteins in these sequences. Sequences are separated by a metabolic pool. A novel feature of these formulations is the translation coefficient, which relates the transport rate attained at given ambient nutrient concentrations and membrane transporter characteristics to the nutrient concentrations sustained in the metabolic pools. The formulations, termed janusian kinetics, show that hyperbolic kinetics are retained during independent changes in transporter and enzyme contents or characteristics. Specific affinity (a degrees (A)) depends strongly on the amount and kinetic characteristics of the transporters; it is also mildly affected by the amount and characteristics of the rate-limiting enzyme. This kinetic constant best describes the ability to accumulate substrate from limiting concentrations. Maximal velocity (V(max)) describes uptake from concentrated solutions and can depend strongly on either limiting enzyme content or the associated content of transporters. The whole-cell Michaelis constant (K(T)), which depends on the ratio of rate-limiting enzyme to transporter, can be relatively independent of change in a degrees (A) and is best used to describe the concentration at which saturation begins to occur. Theory specifies that good oligotrophs have a large a degrees (A) for nutrient collection and a small V(max) for economy of enzyme, giving a small K(T). The product of the two constants is universally rather constant so that oligotrophy is scaled on a plot of a degrees (A) versus K(T), with better oligotrophs toward one end. This idea is borne out by experimental data, and therefore typical small difficult-to-culture aquatic bacteria may be classified as oligobacteria. PMID- 16348525 TI - Growth of Thiobacillus ferrooxidans on Formic Acid. AB - A variety of acidophilic microorganisms were shown to be capable of oxidizing formate. These included Thiobacillus ferrooxidans ATCC 21834, which, however, could not grow on formate in normal batch cultures. However, the organism could be grown on formate when the substrate supply was growth limiting, e.g., in formate-limited chemostat cultures. The cell densities achieved by the use of the latter cultivation method were higher than cell densities reported for growth of T. ferrooxidans on ferrous iron or reduced sulfur compounds. Inhibition of formate oxidation by cell suspensions, but not cell extracts, of formate-grown T. ferrooxidans occurred at formate concentrations above 100 muM. This observation explains the inability of the organism to grow on formate in batch cultures. Cells grown in formate-limited chemostat cultures retained the ability to oxidize ferrous iron at high rates. Ribulose 1,5-bisphosphate carboxylase activities in cell extracts indicated that T. ferrooxidans employs the Calvin cycle for carbon assimilation during growth on formate. Oxidation of formate by cell extracts was NAD(P) independent. PMID- 16348526 TI - Energy Transduction by Anaerobic Ferric Iron Respiration in Thiobacillus ferrooxidans. AB - Formate-grown cells of the obligately chemolithoautotrophic acidophile Thiobacillus ferrooxidans were capable of formate- and elemental sulfur-dependent reduction of ferric iron under anaerobic conditions. Under aerobic conditions, both oxygen and ferric iron could be simultaneously used as electron acceptors. To investigate whether anaerobic ferric iron respiration by T. ferrooxidans is an energy-transducing process, uptake of amino acids was studied. Glycine uptake by starved cells did not occur in the absence of an electron donor, neither under aerobic conditions nor under anaerobic conditions. Uptake of glycine could be driven by formate- and ferrous iron-dependent oxygen uptake. Under anaerobic conditions, ferric iron respiration with the electron donors formate and elemental sulfur could energize glycine uptake. Glycine uptake was inhibited by the uncoupler 2,4-dinitrophenol. The results indicate that anaerobic ferric iron respiration can contribute to the energy budget of T. ferrooxidans. PMID- 16348527 TI - Photocatabolism of Aromatic Compounds by the Phototrophic Purple Bacterium Rhodomicrobium vannielii. AB - The phototrophic purple non-sulfur bacterium Rhodomicrobium vannielii grew phototrophically (illuminated anaerobic conditions) on a variety of aromatic compounds (in the presence of CO(2)). Benzoate was universally photocatabolized by all five strains of R. vannielii examined, and benzyl alcohol was photocatabolized by four of the five strains. Catabolism of benzyl alcohol by phototrophic bacteria has not been previously reported. Other aromatic substrates supporting reasonably good growth of R. vannielii strains were the methoxylated benzoate derivatives vanillate (4-hydroxy-3-methoxybenzoate) and syringate (4 hydroxy-3,5-dimethoxybenzoate). However, catabolism of vanillate and syringate led to significant inhibition of bacteriochlorophyll synthesis in R. vannielii cells, eventually causing cultures to cease growing. No such effect on photopigment synthesis in cells grown on benzoate or benzyl alcohol was observed. Along with a handful of other species of anoxygenic phototrophic bacteria, the ability of the species R. vannielii to photocatabolize aromatic compounds indicates that this organism may also be ecologically significant as a consumer of aromatic derivatives in illuminated anaerobic habitats in nature. PMID- 16348528 TI - Inorganic phosphorus stimulation of bacterioplankton production in a meso eutrophic lake. AB - Experiments were conducted to determine whether production of heterotrophic bacterioplankton in a small meso-eutrophic lake was influenced by the dissolved inorganic phosphorus (DIP) supply. DIP may indirectly limit bacterial production by limiting phytoplankton, which in turn may limit the carbon available to bacteria. Direct DIP limitation of bacteria occurs where the availability of DIP for bacteria is insufficient to maintain growth. This work examined direct DIP limitation of bacteria by removing phytoplankton and incubating flasks with or without added P in the dark. Bacterial production was measured via the rate of incorporation of [H]thymidine ([H]TdR) into DNA. Bacterial abundance was followed with epifluorescent direct counts. Rates of [H]TdR incorporation were significantly greater in flasks with added DIP, and changes in cell abundances generally paralleled increases in [H]TdR incorporation. Even very small additions of P (0.05 muM) were sufficient to stimulate production. DIP addition to whole lakewater also stimulated [H]TdR incorporation relative to that in zero-addition controls, but there was not a concurrent increase in bacterial cell numbers. The stimulation of [H]TdR incorporation after DIP addition to whole lakewater was significantly less than the stimulation due to DIP addition to 1-mum-pore-size filtered lakewater. In this study, addition of DIP caused as much as an eightfold stimulation of [H]TdR incorporation. PMID- 16348529 TI - Hyphomonas spp., Shewanella spp., and Other Marine Bacteria Lack Heterogeneous (Ladderlike) Lipopolysaccharides. AB - The lipopolysaccharides (LPS) of 19 marine bacteria were examined for size heterogeneities by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis in conjunction with an LPS-specific silver staining method. Fifteen marine bacteria had an R-type LPS instead of the ladderlike LPS array characteristic of most bacteria. In addition, three marine bacteria also had a single large LPS molecule. Without constraints (e.g., surface masking), R-type LPS, a more hydrophobic molecule, predominates in Shewanella and Hyphomonas species and in other marine bacteria. PMID- 16348530 TI - Effect of pH on Anaerobic Mild Steel Corrosion by Methanogenic Bacteria. AB - Methanogens can use H(2) produced by cathodic depolarization-mediated oxidation of elemental iron to produce methane. Thermodynamic consideration of the cathodic depolarization mechanism predicts more oxidation of Fe at lower pH. Methanogenic responses to pH by Methanococcus deltae, Methanococcus thermolithotrophicus, and Methanosarcina barkeri were examined. When grown on H(2)-CO(2), these bacteria had pH optima from 6.2 to 7.0, but when all H(2) was supplied from Fe, methanogenic pH optima were lower, 5.4 to 6.5. Corrosion was monitored with and without cultures and at various pHs; more corrosion occurred when cultures were present, biologically induced corrosion was greatest at the pH optima for methanogenesis from Fe, and corrosion without cultures increased with a drop in pH. PMID- 16348531 TI - Purification and Cooperative Activity of Enzymes Constituting the Xylan-Degrading System of Thermomonospora fusca. AB - The thermophilic actinomycete Thermomonospora fusca produced endoxylanase, alpha arabinofuranosidase, beta-xylosidase, and acetyl esterase activities maximally during growth on xylan. Growth yields on glucose, xylose, or arabinose were comparable, but production of endoxylanase and beta-xylosidase was not induced on these substrates. The crude xylanase activity was thermostable and relatively resistant to end product inhibition by xylobiose and xylan hydrolysis products. Six proteins with xylanase activity were identified by zymogram analysis of isoelectric focusing gels, but only a 32-kDa protein exhibiting three isomeric forms could be purified by fast protein liquid chromatography. Endoglucanases were also identified in carboxymethylcellulose-grown cultures, and their distinction from endoxylanases was confirmed. alpha-Arabinofuranosidase activity was due to a single dimeric protein of 92 kDa, which was particularly resistant to end product inhibition by arabinose. Three bands of acetyl esterase activity were detected by zymogram analysis, and there was evidence that these mainly consisted of an intracellular 80-kDa protein secreted to yield active 40-kDa subunits in the culture supernatant. The acetyl esterases were found to be responsible for acetyl xylan esterase activity in T. fusca, in contrast to the distinction proposed in some other systems. The addition of purified betaxylosidase to endoxylanase increased the hydrolysis of xylan, probably by relieving end product inhibition. The enhanced saccharification of wheat straw caused by the addition of purified alpha-arabinofuranosidase to T. fusca endoxylanase suggested a truly synergistic relationship, in agreement with proposals that arabinose side groups on the xylan chain participate in cross linking within the plant cell wall structure. PMID- 16348532 TI - Effects of fungal root pathogens on the population dynamics of biocontrol strains of fluorescent pseudomonads in the wheat rhizosphere. AB - The influences of Gaeumannomyces graminis var. tritici (which causes take-all of wheat), Rhizoctonia solani AG-8 (which causes rhizoctonia root rot of wheat), Pythium irregulare, P. aristosporum, and P. ultimum var. sporangiiferum (which cause pythium root rot of wheat) on the population dynamics of Pseudomonas fluorescens 2-79 and Q72a-80 (bicontrol strains active against take-all and pythium root rot of wheat, respectively) in the wheat rhizosphere were examined. Root infection by either G. graminis var. tritici or R. solani resulted in populations of both bacterial strains that were equal to or significantly larger than their respective populations maintained on roots in the absence of these pathogens. In contrast, the population of strain 2-79 was significantly smaller on roots in the presence of any of the three Pythium species than on noninfected roots and was often below the limits of detection (50 CFU/cm of root) on Pythium infected roots after 40 days of plant growth. In the presence of either P. aristosporum or P. ultimum var. sporangiiferum, the decline in the population of Q72a-80 was similar to that observed on noninfected roots; however, the population of this strain declined more rapidly on roots infected by P. irregulare than on noninfected roots. Application of metalaxyl (which is selectively inhibitory to Pythium spp.) to soil naturally infestated with Pythium spp. resulted in significantly larger rhizosphere populations of the introduced bacteria over time than on plants grown in the same soil without metalaxyl. It is apparent that root infections by fungal pathogens may either enhance or depress the population of fluorescent pseudomonads introduced for their control, with different strains of pseudomonads reacting differentially to different genera and species of the root pathogens. PMID- 16348533 TI - Enhancement of the Potential To Utilize Octopine in the Nonfluorescent Pseudomonas sp. Strain 92. AB - The nonfluorescent Pseudomonas sp. strain 92 requires the presence of a supplementary carbon source for growth on octopine, whereas the spontaneous mutant RB100 has acquired the capacity to utilize this opine as the sole carbon and nitrogen source. Insertional mutagenesis of RB100 with transposon Tn5 generated mutants which were unable to grow on octopine and others which grew slowly on this substrate. Both types of mutants yielded revertants that had regained the ability to utilize octopine. Some of the revertants had lost the transposon, whereas in others the transposon was retained but with rearrangements of the insertion site. Genes of octopine catabolism from strain 92 were cloned on a cosmid vector to generate pK3. The clone pK3 conferred the ability to utilize octopine as the sole carbon and nitrogen source on the host Pseudomonas putida KT2440. Although they conferred an equivalent growth phenotype, the mutant genes carried by RB100 and the cloned genes on pK3 differed in their regulation. Utilization of [C]octopine was inducible by octopine in RB100 and was constitutive in KT2440(pK3). PMID- 16348534 TI - Hydrolysis of Protein and Model Dipeptide Substrates by Attached and Nonattached Marine Pseudomonas sp. Strain NCIMB 2021. AB - Rates of substrate hydrolysis by nonattached bacteria and by bacteria attached to particles derived from marine diatom frustules were estimated by using two substrates, a dipeptide analog and a protein. Adsorption of the two substrates onto the particles was also evaluated. Methyl-coumarinyl-amide-leucine (MCA leucine) was used to estimate hydrolysis of dipeptides by measuring an increase in fluorescence as MCA-leucine was hydrolyzed to leucine and the fluorochrome methylcoumarin. To examine hydrolysis of a larger molecule, we prepared a radiolabeled protein by C-methylation of bovine serum albumin. The rate of protein hydrolysis in samples of particle-attached or nonattached bacteria was estimated by precipitating all nonhydrolyzed protein with cold trichloroacetic acid and then determining the trichloroacetic acid-soluble radiolabeled material, which represented methyl-C-peptides and -amino acids. About 25% of the MCA leucine adsorbed to the particles. MCA-leucine was hydrolyzed faster by nonattached than attached bacteria, which was probably related to its tendency to remain dissolved in the liquid phase. In contrast, almost 100% of the labeled protein adsorbed to the particles. Accordingly, protein was much less available to nonattached bacteria but was rapidly hydrolyzed by attached bacteria. PMID- 16348535 TI - Efficiency of nodule initiation and autoregulatory responses in a supernodulating soybean mutant. AB - We compared the formation of nodules on the primary roots of a soybean cultivar (Glycine max (L.) Merr. cv. Bragg) and a supernodulating mutant derivative, nts382. Inoculation with Bradyrhizobium japonicum USDA 110 at different times after seed imbibition showed that the roots acquired full susceptibility to infection only between 3 and 4 days postgermination. When the plants were inoculated with serial dilutions of a bacterial suspension, the number of nodules formed in the initially susceptible region of the roots was linearly dependent on the logarithm of the inoculum dose until an optimum dose was reached. At least 10 fold-lower doses were required to induce half-maximal nodulation responses on nts382 than on the wild type. However, at optimal doses, about six times as many nodules formed in the initially susceptible region of the roots in nts382. Since there was no appreciable difference in the apparent rates of nodule emergence, the increased efficiency of nodule initiation in the supernodulating mutant could have resulted from a lower threshold of response to bacterial symbiotic signals. Two inoculations (24 h apart) of G. max cv. Bragg revealed that there was a host mediated regulatory response that suppressed nodulation in younger portions of the primary roots, as reported previously for other soybean cultivar Bradyrhizobium combinations. Similar experiments with nts382 revealed a comparable suppressive response, but this response was not as pronounced as it was in the wild type. This and other results suggest that there are additional control mechanisms for nodulation that are different from the systemic autoregulatory control of nodulation altered in supernodulating mutants. PMID- 16348536 TI - Ultrasound treatment for harvesting an aminopeptidase from lactic Acid bacteria and quantitation of the enzyme by enzyme-linked immunosorbent assays. AB - Ultrasound treatment of Lactococcus lactis subsp. cremoris AM2 was optimized to release a maximum amount of intracellular aminopeptidase without modifying the antigenicity of the enzyme. The cells were sonicated three times for 30 s at 23 W. Antibodies produced against the aminopeptidase purified from L. lactis subsp. cremoris AM2 enabled us to use immunoblotting to detect the enzyme in the lysates of all of the lactococci tested but not in the lysates of Leuconostoc strains, lactobacilli, and Streptococcus salivarus subsp. thermophilus. A sandwich enzyme linked immunosorbent assay (ELISA) was developed to quantify the purified aminopeptidase; the detection limit was 4 ng/ml. The aminopeptidase in the supernatant obtained after the ultrasound treatment of strain AM2 cells was detected with the ELISA starting with a total protein concentration of 200 ng/ml. The proportion of equivalent purified aminopeptidase in the supernatant of L. lactis subsp. cremoris AM2 was about 2% of the total protein. Similarly, the aminopeptidase was quantified in different lactococci; the percentages varied between 0.16 and 2%, depending on the strain. The aminopeptidase content in a mixture of several lactic bacteria was also determined with the sandwich ELISA. PMID- 16348537 TI - Anaerobic growth of microorganisms with chlorate as an electron acceptor. AB - The ability of microorganisms to use chlorate (ClO(3)) as an electron acceptor for respiration under anaerobic conditions was studied in batch and continuous tests. Complex microbial communities were cultivated anaerobically in defined media containing chlorate, all essential minerals, and acetate as the sole energy and carbon source. It was shown that chlorate was reduced to chloride, while acetate was oxidized to carbon dioxide and water and used as the carbon source for synthesis of new biomass. A biomass yield of 1.9 to 3.8 g of volatile suspended solids per equivalent of available electrons was obtained, showing that anaerobic growth with chlorate as an electron acceptor gives a high energy yield. This indicates that microbial reduction of chlorate to chloride in anaerobic systems is coupled with electron transport phosphorylation. PMID- 16348538 TI - Spore-forming thermophilic sulfate-reducing bacteria isolated from north sea oil field waters. AB - Thermophilic sulfate-reducing bacteria were isolated from oil field waters from oil production platforms in the Norwegian sector of the North Sea. Spore-forming rods dominated in the enrichments when lactate, propionate, butyrate, or a mixture of aliphatic fatty acids (C(4) through C(6)) was added as a carbon source and electron donor. Representative strains were isolated and characterized. The isolates grew autotrophically on H(2)-CO(2) and heterotrophically on fatty acids such as formate, propionate, butyrate, caproate, valerate, pyruvate, and lactate and on alcohols such as methanol, ethanol, and propanol. Sulfate, sulfite, and thiosulfate but not nitrate could be used as an electron acceptor. The temperature range for growth was 43 to 78 degrees C; the spores were extremely heat resistant and survived 131 degrees C for 20 min. The optimum pH was 7.0. The isolates grew well in salt concentrations ranging from 0 to 800 mmol of NaCl per liter. Sulfite reductase P582 was present, but cytochrome c and desulfoviridin were not found. Electron micrographs revealed a gram-positive cell organization. The isolates were classified as a Desulfotomaculum sp. on the basis of spore formation, general physiological characteristics, and submicroscopic organization. To detect thermophilic spore-forming sulfate-reducing bacteria in oil field water, polyvalent antisera raised against antigens from two isolates were used. These bacteria were shown to be widespread in oil field water from different platforms. The origin of thermophilic sulfate-reducing bacteria in the pore water of oil reservoirs is discussed. PMID- 16348539 TI - Photosynthetic carbon incorporation and turnover in antarctic cryptoendolithic microbial communities: are they the slowest-growing communities on Earth? AB - The main forms of terrestrial life in the cold, desolate Ross Desert of Antarctica are lichen-dominated or cyanobacterium-dominated cryptoendolithic (hidden in rock) microbial communities. Though microbial community biomass (as measured by extractable lipid phosphate) was well within the range of values determined for other microbial communities, community lipid carbon turnover times (calculated from community lipid biomass, rates of community photosynthetic carbon incorporation into lipids versus temperature, and the in situ temperature record) were among the longest on Earth (ca. 20,000 years). When the temperature is above freezing and moisture is present, moderate rates of photosynthesis can be measured. Lichen communities had a psychrophilic temperature response (maximal rate of 4.5 ng of C h m at 10 degrees C) while cyanobacteria communities had maximal rates at 20 to 30 degrees C (3 ng of C h m). These extraordinarily slowly growing communities were not nutrient limited. No significant changes in photosynthetic metabolism were observed upon additions of 100 nM to 1 mM nitrate, ammonium, phosphate, and manganese. These simple, tenacious microbial communities demonstrate strategies of survival under conditions normally considered too extreme for life. PMID- 16348540 TI - Photoadaptation Alters the Ingestion Rate of Paramecium bursaria, a Mixotrophic Ciliate. AB - Bacteriovorous protozoa harboring symbiotic algae are abundant in aquatic ecosystems, yet despite a recent interest in protozoan bacterivory, the influence of light on their ingestion rates has not been investigated. In this study, Paramecium bursaria containing endosymbiotic Chlorella was tested for the effect of light on its ingestion rate. P. bursaria was grown for 4 to 6 days under five different light fluxes ranging from 1 to 90 microeinsteins s m. Ingestion rates were determined by using 0.77-mum-diameter fluorescent microspheres. 4',6 Diamidino-2-phenylindole dihydrochloride-labeled Enterobacter cloacae was used in one experiment to confirm differences in uptake rates of bacteria by P. bursaria. Unlike phagotrophic phytoflagellates, the ciliates demonstrated different ingestion rates in response to different light intensities. Although symbionts contribute carbon to their host via photosynthesis, the paramecia of the present study fed faster after exposure to higher light intensities, whereas their aposymbiotic counterparts (lacking endosymbionts) were unaffected. Light-induced changes in ingestion rates were not immediate, but corresponded to the period of time required for endosymbiont populations to change significantly. This strongly suggests that the symbionts, stimulated by higher light levels, may dictate the feeding rates of their hosts. Thus, light, apart from temperature, may influence the impact of certain protists on natural bacteria and may affect laboratory based determinations of protistan feeding rates. PMID- 16348541 TI - Purification and Properties of a Thermoactive Glucoamylase from Clostridium thermosaccharolyticum. AB - A bacterial glucoamylase was purified from the anaerobic thermophilic bacterium Clostridium thermosaccharolyticum and characterized. The enzyme, which was purified 63-fold, with a yield of 36%, consisted of a single subunit with an apparent molecular mass of 75 kDa. The purified enzyme was able to attack alpha 1,4- and alpha-1,6-glycosidic linkages in various alpha-glucans, liberating glucose with a beta-anomeric configuration. The purified glucoamylase, which was optimally active at 70 degrees C and pH 5.0, attacked preferentially polysaccharides such as starch, glycogen, amylopectin, and maltodextrin. The velocity of oligosaccharide hydrolysis decreased with a decrease in the size of the substrate. The K(m) values for starch and maltose were 18 mg/ml and 20 mM, respectively. Enzyme activity was not significantly influenced by Ca, EDTA, or alpha- or beta-cyclodextrins. PMID- 16348542 TI - Mutation to Wider Virulence in Puccinia graminis f. sp. tritici: Evidence for the Existence of Loci Which Allow the Fungus To Overcome Several Host Stem Rust Resistance Genes Simultaneously. AB - Mutants of Puccinia graminis f. sp. tritici were obtained which were able to overcome simultaneously several host stem rust resistance (Sr) genes effective against the wild-type culture. These results suggest that, in addition to those Psr loci which relate specifically to host Sr genes in a "gene for gene" manner, one or more general loci may be present in this pathogen. The product(s) of these general genes may be necessary for the expression of various host Sr genes. The evolution of a super race capable of overcoming many Sr genes for resistance seems likely, as such a pathogen would not have to give up the many proteins predicted by the gene-for-gene relationship. Moreover, it appears that specificity in the wheat rust system is more complicated than suggested by the gene-for-gene concept. PMID- 16348543 TI - Classification and characterization of heterotrophic microbial communities on the basis of patterns of community-level sole-carbon-source utilization. AB - The BLOLOG redox technology based on tetrazolium dye reduction as an indicator of sole-carbon-source utilization was evaluated as a rapid, community-level method to characterize and classify heterotrophic microbial communities. Direct incubation of whole environmental samples (aquatic, soil, and rhizosphere) in BIOLOG plates containing 95 separate carbon sources produced community-dependent patterns of sole-carbon-source utilization. Principal-component analysis of color responses quantified from digitized images of plates revealed distinctive patterns among microbial habitats and spatial gradients within soil and estuarine sites. Correlation of the original carbon source variables to the principal components gives a functional basis to distinctions among communities. Intensive spatial and temporal analysis of microbial communities with this technique can produce ecologically relevant classifications of heterotrophic microbial communities. PMID- 16348544 TI - Evidence for bacterial chemotaxis to cyanobacteria from a radioassay technique. AB - Lyngbya birgei and Aphanizomenon flos-aquae elicited a significant chemotactic attraction of Aeromonas hydrophila compared with controls lacking cyanobacteria. There was a positive exponential relationship between biomass (chlorophyll a) of L. birgei and A. flos-aquae and chemotactic attraction of A. hydrophila. The assay equipment was simple and reliable and could be used to study bacterial chemotaxis in other species in situ. PMID- 16348546 TI - Evaluation of Four Aerobiological Sampling Methods for the Retrieval of Aerosolized Pseudomonas syringae. AB - [This corrects the article on p. 1268 in vol. 57.]. PMID- 16348545 TI - Amine oxidases of marine phytoplankton. AB - Some phytoplankton utilized a novel mechanism for obtaining nitrogen from primary amines. They oxidized the primary amines to produce extracellular hydrogen peroxide and aldehydes and used the third reaction product, ammonium, as a nitrogen source. The specificity, regulation, inhibition by bromoethylamine, and potential dependence on copper of this process are described. PMID- 16348547 TI - Detection of Escherichia coli and Shigella spp. in Water by Using the Polymerase Chain Reaction and Gene Probes for uid. AB - [This corrects the article on p. 1013 in vol. 57.]. PMID- 16348548 TI - Effects of sulfuroxy anions on degradation of pentachlorophenol by a methanogenic enrichment culture. AB - We studied the degradation of pentachlorophenol (PCP) under methanogenic and sulfate-reducing conditions with an anaerobic mixed culture derived from sewage sludge. The consortium degraded PCP via 2,3,4,5-tetrachlorophenol, 3,4,5 trichlorophenol, and 3,5-dichlorophenol and eventually accumulated 3 chlorophenol. Dechlorination of PCP and metabolites was inhibited in the presence of sulfate, thiosulfate, and sulfite. A decrease in the rate of PCP transformation was noted when the endogenous dissolved H(2) was depleted below 0.11 muM in sulfate-reducing cultures. The effect on dechlorination observed with sulfate could be relieved by addition of molybdate, a competitive inhibitor of sulfate reduction. Addition of H(2) reduced the inhibition observed with sulfuroxy anions. The inhibitory effect of sulfuroxy anions may be due to a competition for H(2) between sulfate reduction and dechlorination. When cultured under methanogenic conditions, the consortium degraded several chlorinated and brominated phenols. PMID- 16348549 TI - Characterization of an Unusual New Agrobacterium tumefaciens Strain from Chrysanthemum morifolium Ram. AB - We characterized five isolates of Agrobacterium tumefaciens from naturally occurring galls on Chrysanthemum morifolium. The isolates are similar, possibly identical, members of a single strain of A. tumefaciens that we designate Chry5. The strain is a biotype I, as indicated by its response to both newly described and traditional biotype tests. Chry5 produces tumors on at least 10 plant species. It is unusual in its ability to form efficiently large tumors on soybean (Glycine max), a species normally refractory to transformation. Chry5 is unable to utilize octopine or mannopine as a carbon source. Although Chry5 can catabolize a single isomer each of nopaline and succinamopine, it differs from other known nopaline and succinamopine strains in its insensitivity to agrocin 84. This pattern of opine catabolism is unique among Agrobacterium strains examined to date. All five isolates of Chry5 contain at least two plasmids, one of which shares homology with pTiB6. PMID- 16348550 TI - Purification and characterization of a pyruvated-mannose-specific xanthan lyase from heat-stable, salt-tolerant bacteria. AB - A xanthanase complex secreted by a consortium of heat-stable, salt-tolerant bacteria includes a lyase that specifically removes terminal pyruvated beta-d mannose residues from the side chains of xanthan gum. The enzyme was purified to homogeneity from the culture broth following ion-exchange chromatography and gel permeation chromatography. It consists of a single subunit of molecular weight 33,000. The enzyme is stable to 55 degrees C for more than 6 h in 20 mM sodium phosphate buffer (pH 5.0) containing 0.25 M NaCl. Optimal enzyme activity was observed at 0.05 M NaCl and a pH of 5. The enzyme has a pI of 3.7. It does not remove unsubstituted terminal beta-d-mannose residues from xanthan side chains nor does it hydrolyze p-nitrophenyl-beta-d-mannose. Treatment of xanthan with purified lyase results in a polysaccharide containing side chains terminating in an unsaturated 4,5-ene-glucuronic acid. PMID- 16348551 TI - Isolation and characterization of microorganisms associated with the traditional sorghum fermentation for production of sudanese kisra. AB - Sorghum flour obtained from Sudan was mixed with water in a 1:2 (wt/vol) ratio and fermented at 30 degrees C for 24 h. The bacterial populations increased with fermentation time and reached a plateau at approximately 18 h. At the end of 24 h, sorghum batter pH had dropped from 5.95 to 3.95 and the batter had a lactic acid content of 0.80%. The microbial population during the 24 h of fermentation consisted of bacteria (Pediococcus pentosaceus, Lactobacillus confusus, Lactobacillus brevis, Lactobacillus sp., Erwinia ananas, Klebsiella pneumoniae, and Enterobacter cloacae), yeasts (Candida intermedia and Debaryomyces hansenii), and molds (Aspergillus sp., Penicillium sp., Fusarium sp., and Rhizopus sp.). P. pentosaceus was the dominant microorganism at the end of the 24-h fermentation. When three consecutive fermentations using an inoculum from the previous fermentation were carried out, the bacterial population increase plateaued at 9 h. The microbial populations in these fermentations were dominated by P. pentosaceus. PMID- 16348552 TI - Autoproteolysis of the Extracellular Serine Proteinase of Lactococcus lactis subsp. cremoris Wg2. AB - The molecular masses of purified extracellular serine proteinase of a number of Lactococcus lactis strains vary significantly, and these molecular mass values do not correspond to the values estimated on the basis of genetic data. The discrepancies can only partially be explained by N-terminal processing during maturation of the precursor enzyme and by C-terminal cleaving during the release from the cell envelope. With a monoclonal antibody that binds in the active site region of the L. lactis proteinase, the processing of the released proteinase was followed. At 30 degrees C the proteinase was degraded with a concomitant loss of beta-casein hydrolytic activity. In the presence of CaCl(2), proteinase degradation was inhibited, and new degradation products were detected. The specific serine proteinase inhibitors phenylmethylsulfonyl fluoride and diisopropylfluorophosphate also inhibited proteinase degradation. Two major high molecular-mass proteinase fragments (165 and 90 kDa) were found to have the same N-terminal amino acid sequence as the mature proteinase, i.e., [Asp-1-Ala-2-Lys-3 Ala-4-Asn-5-Ser-6, indicating that both fragments were formed by cleavage at the C terminus. The N terminus of a proteinase fragment with low molecular mass (58 kDa) started with Gln-215. In this fragment part of the active site region was eliminated, suggesting that it is proteolytically inactive. Unlike larger fragments, this 58-kDa fragment remained intact after prolonged incubations. These results indicate that autoproteolysis of the L. lactis subsp. cremoris Wg2 proteinase ultimately leads to inactivation of the proteinase by deletion of the active site region. PMID- 16348553 TI - Isolation of Light-Enhanced cDNAs of Cercospora kikuchii. AB - Cercospora kikuchii is a fungal pathogen of soybeans which produces a photosensitizing phytotoxic polyketide metabolite, cercosporin. Cercosporin synthesis in culture is modulated by several environmental factors. In addition to the light requirement for toxin action, cercosporin biosynthesis is also highly light regulated. As a first step towards identifying genes involved in cercosporin regulation and biosynthesis, we have used subtractive hybridization to isolate light-enhanced cDNA clones. Six distinct cDNA clones representing genes from a wild-type C. kikuchii strain for which transcript accumulation is positively regulated by light were isolated. To assess the relationship of these light-enhanced cDNAs to cercosporin biosynthesis, we compared corresponding steady-state RNA levels in the wild type and in three mutant strains altered in toxin biosynthesis. Two of the mutant C. kikuchii strains which fail to accumulate cercosporin in response to light also fail to exhibit light-enhanced accumulation of transcripts corresponding to all six light-enhanced cDNAs. Cercosporin accumulation in the third mutant strain, S2, is regulated by medium composition as well as light. S2 fails to accumulate cercosporin in complete medium, a medium which allows significant cercosporin accumulation by the wild type strain. When cultured in complete medium, this mutant strain also fails to show the wild-type, light-enhanced accumulation of transcripts corresponding to five of the six light-enhanced cDNAs. Kinetic analysis demonstrated that transcript accumulation for two of the six light-enhanced cDNAs strongly paralleled cercosporin accumulation in light-grown wild-type culture. PMID- 16348554 TI - Expression of Proteins and Glycoproteins Encoded by the Haploid Nuclei in the Dikaryotic State in the Basidiomycete Agrocybe aegerita. AB - The total proteins and concanavalin A-binding glycoproteins of the cultivated mushroom Agrocybe aegerita were studied in homokaryotic siblings and in dikaryotic strains. The glycoproteins exhibited considerable variability compared with the proteins; the genetic diversity detected in homokaryons in the glycoprotein analysis was 30-fold higher than the genetic diversity revealed by protein analysis, and the glycoprotein patterns could be used to characterize individual genotypes. We found that the expression of glycoproteins in haploid nuclei was significantly asymmetric when the nuclei were paired in dikaryons. The expression levels of the two component nuclei depended on their genotypes, and each haploid nucleus was characterized by its level of expression. Furthermore, some specific glycoproteins that were not detected in all of the homokaryons were newly synthesized in the dikaryotic strains. Among these was a glycoprotein designated gpAa-65, which was identified in all of the dikaryotic strains and appeared to be a good molecular marker of the dikaryotic state. PMID- 16348555 TI - Genomic Relatedness of Xanthomonas campestris Strains Causing Diseases of Citrus. AB - Xanthomonas campestris strains that cause disease in citrus were compared by restriction endonuclease analysis of DNA fragments separated by pulsed-field gel electrophoresis and by DNA reassociation. Strains of X. campestris pv. citrumelo, which cause citrus bacterial spot, were, on average, 88% related to each other by DNA reassociation, although these strains exhibited diverse restriction digest patterns. In contrast, strains of X. campestris pv. citri groups A and B, which cause canker A and canker B, respectively, had relatively homogeneous restriction digest patterns. The groups of strains causing these three different citrus diseases were examined by DNA reassociation and were found to be from 55 to 63% related to one another. Several pathovars of X. campestris, previously shown to cause weakly aggressive symptoms on citrus, ranged from 83 to 90% similar to X. campestris pv. citrumelo by DNA reassociation. The type strain of X. campestris pv. campestris ranged from 30 to 40% similar in DNA reassociation experiments to strains of X. campestris pv. citrumelo and X. campestris pv. citri groups A and B. Whereas DNA reassociation quantified the difference between relatively unrelated groups of bacterial strains, restriction endonuclease analysis distinguished between closely related strains. PMID- 16348556 TI - Abundance of viruses in marine waters: assessment by epifluorescence and transmission electron microscopy. AB - Abundance of bacteria and tiny DNA-associated particles in the upper layer of Japanese coastal and offshore waters was evaluated by epifluorescence microscopy with 0.015-mum-pore-size Nuclepore filters. The number of tiny DNA-associated particles was compared with the abundance of virus particles estimated by transmission electron microscopy. Although a large variation in virus abundance (1.2 x 10 to 35 x 10 ml) was obtained with the transmission electron microscopy method, the ratio of 4',6-diamidino-2-phenylindole-reactive tiny particles to viruses was in a rather narrow range (1.0 to 1.6), indicating that the majority of the tiny DNA-associated particles identified by epifluorescence microscopy were actually virus particles. This result implies the possibility of using epifluorescence microscopy for the evaluation of virus abundance in marine environments. PMID- 16348558 TI - Carbon Concentration and Carbon-to-Nitrogen Ratio Influence Submerged-Culture Conidiation by the Potential Bioherbicide Colletotrichum truncatum NRRL 13737. AB - [This corrects the article on p. 3435 in vol. 56.]. PMID- 16348557 TI - Identification of the proton source for the microbial reductive dechlorination of 2,3,4,5,6-pentachlorobiphenyl. AB - Microbially mediated reductive dechlorination of polychlorinated biphenyls (PCBs) in anaerobic sediments has been observed during laboratory experiments. Reductive dechlorination is a two-electron transfer reaction which involves the release of chlorine as a chloride ion and its replacement on the aromatic ring by hydrogen. The exact mechanism of the electron transfer for PCBs is unknown; however, this work shows that the source of the hydrogen atom is the proton (H) from water. PMID- 16348559 TI - Chromosomal Integration and Expression of Two Bacterial alpha-Acetolactate Decarboxylase Genes in Brewer's Yeast. AB - A bacterial gene encoding alpha-acetolactate decarboxylase, isolated from Klebsiella terrigena or Enterobacter aerogenes, was expressed in brewer's yeast. The genes were expressed under either the yeast phosphoglycerokinase (PGK1) or the alcohol dehydrogenase (ADH1) promoter and were integrated by gene replacement by using cotransformation into the PGK1 or ADH1 locus, respectively, of a brewer's yeast. The expression level of the alpha-acetolactate decarboxylase gene of the PGK1 integrant strains was higher than that of the ADH1 integrants. Under pilot-scale brewing conditions, the alpha-acetolactate decarboxylase activity of the PGK1 integrant strains was sufficient to reduce the formation of diacetyl below the taste threshold value, and no lagering was needed. The brewing properties of the recombinant yeast strains were otherwise unaltered, and the quality (most importantly, the flavor) of the trial beers produced was as good as that of the control beer. PMID- 16348560 TI - Effect of hydrogenase and mixed sulfate-reducing bacterial populations on the corrosion of steel. AB - The importance of hydrogenase activity to corrosion of steel was assessed by using mixed populations of sulfate-reducing bacteria isolated from corroded and noncorroded oil pipelines. Biofilms which developed on the steel studs contained detectable numbers of sulfate-reducing bacteria (10 increasing to 10/0.5 cm). However, the biofilm with active hydrogenase activity (i.e., corrosion pipeline organisms), as measured by a semiquantitative commercial kit, was associated with a significantly higher corrosion rate (7.79 mm/year) relative to noncorrosive biofilm (0.48 mm/year) with 10 sulfate-reducing bacteria per 0.5 cm but no measurable hydrogenase activity. The importance of hydrogenase and the microbial sulfate-reducing bacterial population making up the biofilm are discussed relative to biocorrosion. PMID- 16348561 TI - Propionic Acid Production by a Propionic Acid-Tolerant Strain of Propionibacterium acidipropionici in Batch and Semicontinuous Fermentation. AB - A propionic acid-tolerant derivative of Propionibacterium acidipropionici P9 was obtained by serially transferring strain P9 through broth that contained increasing amounts of propionic acid. After 1 year of repeated transfers, a strain (designed P200910) capable of growth at higher propionic acid concentrations than P9 was obtained. An increase in the proportion of cellular straight-chain fatty acids and uncoupling of propionic acid production from growth were observed for strain P200910. Growth rate, sugar utilization, and acid production were monitored during batch and semicontinuous fermentations of semidefined medium and during batch fermentation of whey permeate for both strain P200910 and strain P9. The highest propionic acid concentration (47 g/liter) was produced by P200910 in a semicontinuous fermentation. Strain P200910 produced a higher ratio of propionic acid to acetic acid, utilized sugar more efficiently, and produced more propionic acid per gram of biomass than did its parent in all fermentations. PMID- 16348562 TI - Method to enhance growth and sporulation of pelletized biocontrol fungi. AB - The biocontrol fungi Trichoderma harzianum, used to control soilborne plant pathogens, and Beauveria bassiana, used to control insect pests, were formulated as mycelial biomass in alginate pellets with wheat bran added. After drying for 0, 4, or 16 h, pellets were placed in water or in aqueous solutions of polyethylene glycol (PEG) 8000 for 4 to 24 h and then allowed to continue drying. PEG-treated pellets containing T. harzianum showed significantly greater proliferation of hyphae in soil than untreated pellets or pellets treated with water. Production of conidia of T. harzianum from PEG-treated pellets was lower than production from untreated pellets after 4 days, although rates were equivalent after 7 days. In contrast, production of conidia of B. bassiana was significantly more rapid from PEG-treated pellets than from untreated pellets. Biocontrol of soilborne plant pathogens or insect pests may be enhanced by rapid hyphal growth of T. harzianum in soil or rapid sporulation of B. bassiana on foliage, respectively. Therefore, PEG treatment may improve the efficacy of these biocontrol agents. PMID- 16348563 TI - Proteases and Their Involvement in the Infection and Immobilization of Nematodes by the Nematophagous Fungus Arthrobotrys oligospora. AB - The nematophagous fungus Arthrobotrys oligospora produced extracellular proteases when grown in a liquid culture, as revealed by measuring the hydrolysis of the chromogenic substrate Azocoll. The extracellular protease activity was inhibited by phenylmethylsulfonyl fluoride (PMSF) and other serine protease inhibitors and partly inhibited by the aspartate protease inhibitor pepstatin and by a cysteine protease inhibitor [l-trans-epoxysuccinyl-leucylamide-(4-guanidino)-butane, or E 64]. Substrate gel electrophoresis showed that the fungus produced several different proteases, including multiple serine proteases. The function of proteases in the infection of nematodes was examined by treating the fungus with various protease inhibitors. None of the inhibitors tested affected the adhesion of nematodes to the traps, but incubating trap-bearing mycelium with a serine protease inhibitor, PMSF, antipain, or chymostatin, or the metalloprotease inhibitor phenanthroline significantly decreased the immobilization of nematodes captured by the fungus. Inhibitors of cysteine or aspartic proteases did not affect the immobilization of captured nematodes. The effects of PMSF on the immobilization of nematodes were probably due to serine proteases produced by the fungus, since the effects were observed when unbound inhibitor was washed away from the fungus before the nematodes were added to the system. No effects were observed when the nematodes only were pretreated with PMSF. PMID- 16348564 TI - Self-Cycling Fermentation Applied to Acinetobacter calcoaceticus RAG-1. AB - The self-cycling fermentation (SCF) technique was applied to a culture of Acinetobacter calcoaceticus RAG-1. This method was shown to result in synchronization of the cells, achieving a 77% improvement in cell synchrony over that of the batch case. Cellular occurrences, averaged out by asynchronous batch cultures, were magnified by the temporal alignment of metabolic events brought about by the synchronization associated with SCFs. The cell population doubled only once per cycle, thus establishing an equality between cycle time and doubling time. Parameters of interest were biomass concentration, total bioemulsifier (emulsan) production, cycle time, and residual carbon concentration. Cycle-to-cycle variation of these parameters was, in most cases, insignificant. Repeatability of doubling time estimates (based on 95% confidence intervals) was roughly 7 to 10 times better between cycles in an SCF than between batch replicates. The carbon substrate was completely utilized in all cases in which it was measured, giving this technique an advantage over chemostat-type fermentations. The dissolved-oxygen profiles monitored throughout a cycle were found to be repeatable. A characteristic shape, which can be related to the growth of the organism, was associated with each carbon source. The specific emulsan productivity of SCFs was found to be approximately 50 times greater than that of the batch process and 2 to 9 times greater than that of the chemostat, depending on the dilution rate considered. With respect to specific emulsan production, a 25-fold improvement over that in an immobilized cell system recently introduced was obtained. Thus, SCFs are a viable alternative to established fermentation techniques. PMID- 16348565 TI - Chemotaxis of Pseudomonas syringae subsp. savastanoi Virulence Mutants. AB - Several mutants of Pseudomonas syringae subsp. savastanoi were tested for their ability to sense and respond to a chemotactic gradient in low concentrations of yeast extract. The mutants were deficient in one or both of the genes coding for the synthesis of the plant hormones indole-3-acetic acid (IAA) and isopentenyl adenosine. Mutations which resulted in the loss of IAA production were due to the loss of the entire plasmid containing the iaa operon or to an 18-kb deletion of the iaa region. Additional mutants tested were deficient in their ability to produce isopentenyl adenosine as a result of the loss of the ptz-bearing plasmid. In all cases, strains which had lost the ability to produce IAA exhibited enhanced motility of up to 2.5 times that of the wild type (IAA) in medium containing 0.01% yeast extract. No differences in motility were observed on medium containing lower concentrations of yeast extract. The presence or absence of the cytokinin plasmid and the presence or absence of inorganic nitrogen in the medium had no effect on the relative mobility of the strains. PMID- 16348566 TI - Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii. AB - An altered beta-tubulin gene that confers resistance to the fungicide benomyl was isolated from a genomic library of a UV-induced mutant of Cercospora kikuchii and used as a selectable marker for transformation. The level of benomyl resistance conferred to the transformants was at least 150-fold greater than the intrinsic resistance of the C. kikuchii recipient protoplasts. In the majority of cases, the tubulin fragment was integrated at the native beta-tubulin locus, apparently by gene replacement or gene conversion. The frequency of transformation ranged from 0.2 to 6 transformants per mug of DNA, depending on the recipient strain. Transformation with linearized plasmid resulted in a higher frequency, without changing the type of integration event. Transformants were phenotypically stable after eight consecutive transfers on medium without benomyl. This is the first report of a genetic transformation system for a Cercospora species. PMID- 16348567 TI - Mutants of Cercospora kikuchii Altered in Cercosporin Synthesis and Pathogenicity. AB - We have obtained spontaneous and UV-induced stable mutants, altered in the synthesis of cercosporin, of the fungal soybean pathogen Cercospora kikuchii. The mutants were isolated on the basis of colony color on minimal medium. The UV induced mutants accumulated, at most, 2% of wild-type cercosporin levels on all media tested. In contrast, cercosporin accumulation by the spontaneous mutants was strongly medium regulated, occurring only on potato dextrose medium but at concentrations comparable to those produced by the wild-type strain. UV-induced mutants unable to synthesize cercosporin on any medium were unable to incite lesions when inoculated onto the soybean host. Cercosporin was reproducibly isolated from all inoculated leaves showing lesions. Although cercosporin involvement in disease has been indirectly suggested by many previous studies, this is the first report in which mutants blocked in cercosporin synthesis have been used to demonstrate that cercosporin is a crucial pathogenicity factor for this fungal genus. PMID- 16348568 TI - Factors Limiting Aliphatic Chlorocarbon Degradation by Nitrosomonas europaea: Cometabolic Inactivation of Ammonia Monooxygenase and Substrate Specificity. AB - The soil nitrifying bacterium Nitrosomonas europaea is capable of degrading trichloroethylene (TCE) and other halogenated hydrocarbons. TCE cometabolism by N. europaea resulted in an irreversible loss of TCE biodegradative capacity, ammonia-oxidizing activity, and ammonia-dependent O(2) uptake by the cells. Inactivation was not observed in the presence of allylthiourea, a specific inhibitor of the enzyme ammonia monooxygenase, or under anaerobic conditions, indicating that the TCE-mediated inactivation required ammonia monooxygenase activity. When N. europaea cells were incubated with [C]TCE under conditions which allowed turnover of ammonia monooxygenase, a number of cellular proteins were covalently labeled with C. Treatment of cells with allylthiourea or acetylene prior to incubation with [C]TCE prevented incorporation of C into proteins. The ammonia-oxidizing activity of cells inactivated in the presence of TCE could be recovered through a process requiring de novo protein synthesis. In addition to TCE, a series of chlorinated methanes, ethanes, and other ethylenes were screened as substrates for ammonia monooxygenase and for their ability to inactivate the ammonia-oxidizing system of N. europaea. The chlorocarbons could be divided into three classes depending on their biodegradability and inactivating potential: (i) compounds which were not biodegradable by N. europaea and which had no toxic effect on the cells; (ii) compounds which were cooxidized by N. europaea and had little or no toxic effect on the cells; and (iii) compounds which were cooxidized and produced a turnover-dependent inactivation of ammonia oxidation by N. europaea. PMID- 16348569 TI - NH(4)-Excreting Azospirillum brasilense Mutants Enhance the Nitrogen Supply of a Wheat Host. AB - Spontaneous ethylenediamine-resistant mutants of Azospirillum brasilense were selected on the basis of their excretion of NH(4). Two mutants exhibited no repression of their nitrogenase enzyme systems in the presence of high (20 mM) concentrations of NH(4). The nitrogenase activities of these mutants on nitrogen free minimal medium were two to three times higher than the nitrogenase activity of the wild type. The mutants excreted substantial amounts of ammonia when they were grown either under oxygen-limiting conditions (1 kPa of O(2)) or aerobically on nitrate or glutamate. The mutants grew well on glutamate as a sole nitrogen source but only poorly on NH(4)Cl. Both mutants failed to incorporate [C]methylamine. We demonstrated that nitrite ammonification occurs in the mutants. Wild-type A. brasilense, as well as the mutants, became established in the rhizospheres of axenically grown wheat plants at levels of > 10 cells per g of root. The rhizosphere acetylene reduction activity was highest in the preparations containing the mutants. When plants were grown on a nitrogen-free nutritional medium, both mutants were responsible for significant increases in root and shoot dry matter compared with wild-type-treated plants or with noninoculated controls. Total plant nitrogen accumulation increased as well. When they were exposed to a N(2)-enriched atmosphere, both A. brasilense mutants incorporated significantly higher amounts of N inside root and shoot material than the wild type did. The results of our nitrogen balance and N enrichment studies indicated that NH(4)-excreting A. brasilense strains potentially support the nitrogen supply of the host plants. PMID- 16348571 TI - Influence of Potassium in the Agar Medium on the Growth Pattern of the Filamentous Fungus Fusarium solani. AB - A decrease in the concentration of K ions below 3 mM in agar medium which also contained starch, casein hydrolysate, MgSO(4), and K(2)HPO(4) changed the growth pattern of Fusarium solani illuminated in diurnal 12-h light/12-h dark cycles from zonation to a feathery growth mode. Rubidium or cesium ions could replace potassium, but lithium, sodium, and the bivalent alkaline earth ions could not. PMID- 16348570 TI - Influence of CO(2)-HCO(3) Levels and pH on Growth, Succinate Production, and Enzyme Activities of Anaerobiospirillum succiniciproducens. AB - Growth and succinate versus lactate production from glucose by Anaerobiospirillum succiniciproducens was regulated by the level of available carbon dioxide and culture pH. At pH 7.2, the generation time was almost doubled and extensive amounts of lactate were formed in comparison with growth at pH 6.2. The succinate yield and the yield of ATP per mole of glucose were significantly enhanced under excess-CO(2)-HCO(3) growth conditions and suggest that there exists a threshold level of CO(2) for enhanced succinate production in A. succiniciproducens. Glucose was metabolized via the Embden-Meyerhof-Parnas route, and phosphoenolpyruvate carboxykinase levels increased while lactate dehydrogenase and alcohol dehydrogenase levels decreased under excess-CO(2)-HCO(3) growth conditions. Kinetic analysis of succinate and lactate formation in continuous culture indicated that the growth rate-linked production rate coefficient (K) cells was much higher for succinate (7.2 versus 1.0 g/g of cells per h) while the non-growth-rate-related formation rate coefficient (K') was higher for lactate (1.1 versus 0.3 g/g of cells per h). The data indicate that A. succiniciproducens, unlike other succinate-producing anaerobes which also form propionate, can grow rapidly and form high final yields of succinate at pH 6.2 and with excess CO(2)-HCO(3) as a consequence of regulating electron sink metabolism. PMID- 16348572 TI - Differences between Lactobacillus casei subsp. casei 2206 and Citrate-Positive Lactococcus lactis subsp. lactis 3022 in the Characteristics of Diacetyl Production. AB - Lactobacillus casei subsp. casei 2206 exhibited much lower levels of diacetyl reductase activity than CitrLactococcus lactis subsp. lactis 3022 but two-, three , and more than eightfold-higher levels of diacetyl synthase, lactate dehydrogenase, and NADH oxidase activities, respectively. A requirement for metal ions by the diacetyl synthases in both species was observed. The extracts of strain 2206 but not strain 3022 produced more diacetyl from pyruvate when the reaction for diacetyl synthase was aerated than when it was conducted statically. PMID- 16348573 TI - Use of immunogold labelling with scanning electron microscopy to identify phytopathogenic bacteria on leaf surfaces. AB - Scanning electron microscopy coupled with high-resolution back-scattered electron imaging was used to detect gold-labelled specific immunoglobulins attached to epiphytic bacteria. Strains of Xanthomonas citri and X. campestris pv. citrumelo were specifically identified on grapefruit leaf surfaces when labelled with homologous gold-labelled immunoglobulins. PMID- 16348574 TI - Reverse sample genome probing, a new technique for identification of bacteria in environmental samples by DNA hybridization, and its application to the identification of sulfate-reducing bacteria in oil field samples. AB - A novel method for the identification of bacteria in environmental samples by DNA hybridization is presented. It is based on the fact that, even within a genus, the genomes of different bacteria may have little overall sequence homology. This allows the use of the labeled genomic DNA of a given bacterium (referred to as a "standard") to probe for its presence and that of bacteria with highly homologous genomes in total DNA obtained from an environmental sample. Alternatively, total DNA extracted from the sample can be labeled and used to probe filters on which denatured chromosomal DNA from relevant bacterial standards has been spotted. The latter technique is referred to as reverse sample genome probing, since it is the reverse of the usual practice of deriving probes from reference bacteria for analyzing a DNA sample. Reverse sample genome probing allows identification of bacteria in a sample in a single step once a master filter with suitable standards has been developed. Application of reverse sample genome probing to the identification of sulfate-reducing bacteria in 31 samples obtained primarily from oil fields in the province of Alberta has indicated that there are at least 20 genotypically different sulfate-reducing bacteria in these samples. PMID- 16348575 TI - Determinants encoding resistance to several heavy metals in newly isolated copper resistant bacteria. AB - Three copper-resistant, gram-negative bacteria were isolated and characterized. Of the three strains, Alcaligenes denitrificans AH tolerated the highest copper concentration (MIC = 4 mM CuSO(4)). All three strains showed various levels of resistance to other metal ions. A. denitrificans AH contains sequences which cross-hybridized with the mer (mercury resistance) determinant of Tn21 and the czc (cobalt, zinc, and cadmium resistance), cnr (cobalt and nickel resistance), and chr (chromate resistance) determinants of A. eutrophus CH34. DNA-DNA hybridization with probes prepared from A. eutrophus CH34 and Tn21 revealed the presence of chr-, cnr-, and mer-like sequences on the 200-kb plasmid pHG27 and of czc, cnr, and mer homologs located on the chromosome. The second strain, classified as Alcaligenes sp. strain PW, carries czc, cnr, and mer homologs on the 240-kb plasmid pHG29-c and a chr determinant on the 290-kb plasmid pHG29-a; a third plasmid, the 260-kb large plasmid pHG29-b, is cryptic. In contrast to the Alcaligenes strains, which were isolated from metal-contaminated water, Pseudomonas paucimobilis CD was isolated from the air. This strain harbors two cryptic plasmids: the 210-kb large plasmid pHG28-a and the 40-kb plasmid pHG28-b. Southern analysis revealed no homology between the metal ion resistance determinants of A. eutrophus CH34 and P. paucimobilis CD. PMID- 16348576 TI - Contemporaneous N(2) Fixation and Oxygenic Photosynthesis in the Nonheterocystous Mat-Forming Cyanobacterium Lyngbya aestuarii. AB - The nonheterocystous filamentous cyanobacterial genus Lyngbya is a widespread and frequently dominant component of marine microbial mats. It is suspected of contributing to relatively high rates of N(2) fixation associated with mats. The ability to contemporaneously conduct O(2)-sensitive N(2) fixation and oxygenic photosynthesis was investigated in Lyngbya aestuarii isolates from a North Carolina intertidal mat. Short-term (<4-h) additions of the photosystem II (O(2) evolution) inhibitor 3(3,4-dichlorophenyl)-1,1-dimethylurea stimulated light mediated N(2) fixation (nitrogenase activity), indicating potential inhibition of N(2) fixation by O(2) production. However, some degree of light-mediated N(2) fixation in the absence of 3(3,4-dichlorophenyl)-1,1-dimethylurea was observed. Electron microscopic immunocytochemical localization of nitrogenase, coupled to microautoradiographic studies of CO(2) fixation and cellular deposition of the tetrazolium salt 2,4,5-triphenyltetrazolium chloride, revealed that (i) nitrogenase was widely distributed throughout individual filaments during illuminated and dark periods, (ii) CO(2) fixation was most active in intercalary regions, and (iii) daylight 2,4,5-triphenyltetrazolium chloride reduction (formazan deposition) was most intense in terminal regions. Results suggest lateral partitioning of photosynthesis and N(2) fixation during illumination, with N(2) fixation being confined to terminal regions. During darkness, a larger share of the filament appears capable of N(2) fixation. PMID- 16348577 TI - Increased bacterial uptake of macromolecular substrates with fluid shear. AB - To investigate the effect of fluid shear on uptake rates of low-diffusivity macromolecular substrates by suspended cultures, we measured the uptake of two compounds as models of macromolecules, a protein (bovine serum albumin [BSA]) and a polysaccharide (dextran), using pure cultures of Zoogloea ramigera and Escherichia coli, respectively. Oxygen utilization rates of stirred samples grown on BSA and dextran were 2.3 and 2.9 times higher, respectively, than those of undisturbed (still) samples. Uptake rates of H-BSA and [H]dextran by stirred samples were 12.6 and 6.2 times higher, respectively, than those by still samples. These experimentally obtained increases are larger than those predicted with a mass transfer model. Model results indicated that stirring would increase uptake by factors of 1.6 and 1.8 for BSA and dextran. As predicted by the model, we also found that uptake rates of low-molecular-weight substrates with high diffusivities, such as leucine and glucose, were only slightly affected by fluid shear. Since macromolecules can make up a major portion of bacterial substrate in natural, laboratory, and engineered systems, the demonstrated effect of fluid shear has wide implications for kinetic studies performed in basic metabolic research as well as in the evaluation of engineered bioreactors used for wastewater treatment. PMID- 16348578 TI - Differential fermentation of cellulose allomorphs by ruminal cellulolytic bacteria. AB - In addition to its usual native crystalline form (cellulose I), cellulose can exist in a variety of alternative crystalline forms (allomorphs) which differ in their unit cell dimensions, chain packing schemes, and hydrogen bonding relationships. We prepared, by various chemical treatments, four different alternative allomorphs, along with an amorphous (noncrystalline) cellulose which retained its original molecular weight. We then examined the kinetics of degradation of these materials by two species of ruminal bacteria and by inocula from two bovine rumens. Ruminococcus flavefaciens FD-1 and Fibrobacter succinogenes S85 were similar to one another in their relative rates of digestion of the different celluloses, which proceeded in the following order: amorphous > III(I) > IV(I) > III(II) > I > II. Unlike F. succinogenes, R. flavefaciens did not degrade cellulose II, even after an incubation of 3 weeks. Comparisons of the structural features of these allomorphs with their digestion kinetics suggest that degradation is enhanced by skewing of adjacent sheets in the microfibril, but is inhibited by intersheet hydrogen bonding and by antiparallelism in adjacent sheets. Mixed microflora from the bovine rumens showed in vitro digestion rates quite different from one another and from those of both of the two pure bacterial cultures, suggesting that R. flavefaciens and F. succinogenes (purportedly among the most active of the cellulolytic bacteria in the rumen) either behave differently in the ruminal ecosystem from the way they do in pure culture or did not play a major role in cellulose digestion in these ruminal samples. PMID- 16348579 TI - Carbohydrate signatures of aquatic macrophytes and their dissolved degradation products as determined by a sensitive high-performance ion chromatography method. AB - The sugar contents of emergent macrophytes from a freshwater lake, a freshwater swamp, and a salt marsh in the southeastern United States were examined together with the dissolved free sugars produced during macrophyte degradation and in natural water samples collected adjacent to macrophyte stands. Simultaneous separation of up to 13 neutral and 2 amino sugars together with 3 uronic acids and muramic acid was achieved by anion-exchange high-performance ion chromatography. As little as 10 pmol or a concentration of 20 nM sugar can be detected by pulsed amperometry, a greater sensitivity for sugar quantification than that of previously reported detection techniques used in conjunction with either gas or liquid chromatographic systems. Optimum conditions for hydrolysis of plant material by using trifluoroacetic acid were determined, and internal standards were used to quantify losses due to matrix effects and solid-phase extraction of samples. Our data demonstrate that ratios of certain indicator sugars in undegraded macrophytes differ significantly from ratios of dissolved free sugars formed during macrophyte degradation, reflecting the complex processes (biological and physical) involved in vascular plant degradation in aquatic ecosystems. Natural water samples collected adjacent to macrophyte beds contained dissolved free sugars at concentrations of 620 nM (lake), 890 nM (freshwater swamp), and 2,300 nM (salt marsh). Sugar signatures of these natural water samples were similar to those of macrophyte degradation products. PMID- 16348580 TI - Horizontal variation of bacterioplankton in the baltic sea. AB - Variations in the thymidine incorporation rate, bacterial abundance, and mean cell volumes in the surface water (0.5 m) of the Baltic Sea in spring and summer were compared in studies with different spatial scales (570 nautical miles [nmi] [ca. 1056 km], 220 nmi [ca. 407 km], 24 nmi [ca. 44 km], 12 nmi [ca. 22 km], and 200 m). The objective of the comparison was to investigate whether a single sample taken at one sampling point is representative enough for researchers to make generalizations about a larger water area. Bacterioplankton variation was connected more to seasonal characteristics than to the spatial scale of sampling. Variation was greater and more random in spring than in summer. The state variables (bacterial abundance and mean cell volume) varied less than the rate variable (thymidine incorporation). The results suggest that the sampling design for bacterioplankton studies in northern temperate seas should be planned primarily according to the season and that more stress should be put on rate variable measurements than on those of state variables. PMID- 16348581 TI - Visualization of Trichoderma reesei Cellobiohydrolase I and Endoglucanase I on Aspen Cellulose by Using Monoclonal Antibody-Colloidal Gold Conjugates. AB - Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH I) and endoglucanase I (EG I) were conjugated to 10- and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I was visualized by utilizing the MAb colloidal gold probes. The visualization procedure involved immobilization of cellulose microfibrils on copper electron microscopy grids, incubation of the cellulose-coated grids with cellulase(s), binding of MAb-colloidal gold conjugates to cellulase(s), and visualization via transmission electron microscopy. CBH I was seen bound to apparent crystalline cellulose as well as apparent amorphous cellulose. EG I was seen bound extensively to apparent amorphous cellulose with minimal binding to crystalline cellulose. PMID- 16348582 TI - Attachment, Chemotaxis, and Multiplication of Agrobacterium tumefaciens Biovar 1 and Biovar 3 on Grapevine and Pea. AB - Tumorigenic (CG49) and nontumorigenic (CG484) strains of Agrobacterium tumefaciens bv. 3 attached to grape roots at a higher level than did a nonpectinolytic mutant of CG49 (CG50) or a tumorigenic strain of A. tumefaciens bv. 1 (CG628). Strains attached equally well to wounded and unwounded grape roots. Strains responded differently to pea plants in that biovar 3 strains consistently attached to unwounded roots at a lower level than they did to wounded roots, whereas CG628 attached equally well regardless of wounding. The lowest levels of attachment to pea roots were consistently observed for CG50. Population curves were calculated for the strains inoculated into wound sites on grape and pea roots. A. tumefaciens bv. 3 wild-type strains developed greater populations at wound sites on grape roots after 100 h (resulting in root decay) than did CG50 or CG628. Population curves for strains at wound sites on pea roots were different from those on grape roots. There were no significant differences in populations after 100 h, and no strains caused root decay. No differences in the chemotaxis of wild-type and mutant A. tumefaciens bv. 3 strains towards grape roots, crown pieces, or root extracts were observed, but the biovar 1 strain, CG628, always migrated the greatest distance towards all substrates. Polygalacturonase production may affect attachment to grape roots and multiplication of A. tumefaciens bv. 3 at wound sites and thus be associated with the specificity of the bacterium for grape. PMID- 16348583 TI - Hydrophobicity, adhesion, and surface-exposed proteins of gliding bacteria. AB - The cell surface hydrophobicities of a variety of aquatic and terrestrial gliding bacteria were measured by an assay of bacterial adherence to hydrocarbons (BATH), hydrophobic interaction chromatography, and the salt aggregation test. The bacteria demonstrated a broad range of hydrophobicities. Results among the three hydrophobicity assays performed on very hydrophilic strains were quite consistent. Bacterial adhesion to glass did not correlate with any particular measure of surface hydrophobicity. Several adhesion-defective mutants of Cytophaga sp. strain U67 were found to be more hydrophilic than the wild type, particularly by the BATH assay and hydrophobic interaction chromatography. The very limited adhesion of these mutants correlated well with hydrophilicity as determined by the BATH assay. The hydrophobicities of several adhesion-competent revertants ranged between those of the wild type and the mutants. As measured by the BATH assay, starvation increased hydrophobicity of both the wild type and an adhesion-defective mutant. During filament fragmentation of Flexibacter sp. strain FS-1, marked changes in hydrophobicity and adhesion were accompanied by changes in the arrays of surface-exposed proteins as detected by an immobilized radioiodination procedure. PMID- 16348584 TI - Effects of genetically engineered microorganisms on nitrogen transformations and nitrogen-transforming microbial populations in soil. AB - The principal concern about releasing genetically engineered microorganisms (GEMs) into the environment is their potential adverse effects on the environment, whether caused directly or indirectly by the GEMs. The effects of five GEMs on ammonification, nitrification, and denitrification in soil were studied. With the possible exception of a strain of Enterobacter cloacae carrying a plasmid, no consistent statistically or ecologically significant differences in effects on these processes or on the population dynamics of the microorganisms responsible for the processes were observed between soils inoculated with the GEMs or their homologous plasmidless hosts and those that were not inoculated. Increasing the concentration of montmorillonite in the soil enhanced the rate of nitrification, regardless of the inoculum, indicating that the perfusion technique used was sensitive enough to detect changes in nitrification rates when they occurred. PMID- 16348585 TI - Oxygen Uptake and Hydrogen-Stimulated Nitrogenase Activity from Azorhizobium caulinodans ORS571 Grown in a Succinate-Limited Chemostat. AB - Succinate-limited continuous cultures of an Azorhizobium caulinodans strain were grown on ammonia or nitrogen gas as a nitrogen source. Ammonia-grown cells became oxygen limited at 1.7 muM dissolved oxygen, whereas nitrogen-fixing cells remained succinate limited even at dissolved oxygen concentrations as low as 0.9 muM. Nitrogen-fixing cells tolerated dissolved oxygen concentrations as high as 41 muM. Succinate-dependent oxygen uptake rates of cells from the different steady states ranged from 178 to 236 nmol min mg of protein and were not affected by varying chemostat-dissolved oxygen concentration or nitrogen source. When equimolar concentrations of succinate and beta-hydroxybutyrate were combined, oxygen uptake rates were greater than when either substrate was used alone. Azide could also used alone as a respiratory substrate regardless of nitrogen source; however, when azide was added following succinate additions, oxygen uptake was inhibited in ammonia-grown cells and stimulated in nitrogen-fixing cells. Use of 25 mM succinate in the chemostat resevoir at a dilution rate of 0.1 h resulted in high levels of background respiration and nitrogenase activity, indicating that the cells were not energy limited. Lowering the reservoir succinate to 5 mM imposed energy limitation. Maximum succinate-dependent nitrogenase activity was 1,741 nmol of C(2)H(4)h mg (dry weight), and maximum hydrogen-dependent nitrogenase activity was 949 nmol of C(2)H(4) h mg (dry weight). However, when concentration of 5% (vol/vol) hydrogen or greater were combined with succinate, nitrogenase activity decreased by 35% in comparison to when succinate was used alone. Substitution of argon for nitrogen in the chemostat inflow gas resulted in "washout," proving that ORS571 can grow on N(2) and that there was not a nitrogen source in the medium that could substitute. PMID- 16348586 TI - Production and consumption of dimethylsulfoniopropionate in marine microbial mats. AB - The fate of dimethylsulfoniopropionate (DMSP), a major sulfonium compound in marine ecosystems, was examined in Microcoleus chthonoplastes-dominated microbial mats. Chemical decomposition of DMSP was observed under laboratory conditions at pH values higher than 10.0. pH profiles measured in situ showed that these highly alkaline conditions occurred in microbial mats. Axenic cultures of M. chthonoplastes contained 37.3 mumol of DMSP g of protein, which was partially liberated when the cells were subjected to an osmotic shock. DMSP-amended mat slurries showed a rapid turnover of this compound. The addition of glutaraldehyde blocked DMSP decrease, indicating biological consumption. Populations of potential dimethyl sulfide consumers were found in the top 10 mm of the mat. PMID- 16348587 TI - Alteration of the Magnetic Properties of Aquaspirillum magnetotacticum by a Pulse Magnetization Technique. AB - The presence of a narrow shape and size distribution for magnetite crystals within magnetotactic organisms suggests strongly that there are species-specific mechanisms that control the process of biomineralization. In order to explore the extent of this control, cultures of Aquaspirillum magnetotacticum in the exponential growth phase were exposed to increasing magnetic pulses with the aim of separating cell populations on the basis of their magnetic coercivities. Isothermal remanent magnetization and anhysteretic remanent magnetization studies were performed with freeze-dried magnetic cells after the remagnetization treatment. Subpopulations of A. magnetotacticum that showed an increase in coercivity correlated with the intensity of the magnetic pulses were isolated. After successive subcultures of the remaining north-seeking cells, a maximum bulk coercivity (H(b)) of 40 mT was obtained after treatment with a 55-mT pulse. Although we obtained A. magnetotacticum variants displaying higher coercivities than the wild-type strain, changes in crystal size or shape of the magnetite crystals were below reliable detection limits with transmission electron microscopy. Attempts to shift the coercivity towards higher values caused it to decrease, a change which was accompanied by an increase in magnetostatic interactions of the magnetosome chains as well as an increase in the cell population displaying an abnormal distribution of the magnetosome chains. Ultrastructural analyses of cells and magnetosomes revealed the appearance of cystlike bodies which occasionally contained magnetosomes. The increase in cystlike cells and abnormal magnetosome chains when higher magnetic pulses were used suggested that magnetosomes were collapsing because of stronger interparticle magnetostatic forces. PMID- 16348588 TI - Competition for Ammonium between Nitrifying and Heterotrophic Bacteria in Dual Energy-Limited Chemostats. AB - The absence of nitrification in soils rich in organic matter has often been reported. Therefore, competition for limiting amounts of ammonium between the chemolithotrophic ammonium-oxidizing species Nitrosomonas europaea and the heterotrophic species Arthrobacter globiformis was studied in the presence of Nitrobacter winogradskyi in continuous cultures at dilution rates of 0.004 and 0.01 h. Ammonium limitation of A. globiformis was achieved by increasing the glucose concentration in the reservoir stepwise from 0 to 5 mM while maintaining the ammonium concentration at 2 mM. The numbers of N. europaea and N. winogradskyi cells decreased as the numbers of heterotrophic bacteria rose with increasing glucose concentrations for both dilution rates. Critical carbon-to nitrogen ratios of 11.6 and 9.6 were determined for the dilution rates of 0.004 and 0.01 h, respectively. Below these critical values, coexistence of the competing species was found in steady-state situations. Although the numbers were strongly reduced, the nitrifying bacteria were not fully outcompeted by the heterotrophic bacteria above the critical carbon-to-nitrogen ratios. Nitrifying bacteria could probably maintain themselves in the system above the critical carbon-to-nitrogen ratios because they are attached to the glass wall of the culture vessels. The numbers of N. europaea decreased more than did those of N. winogradskyi. This was assumed to be due to heterotrophic growth of the latter species on organic substrates excreted by the heterotrophic bacteria. PMID- 16348589 TI - Transfer of the Pea Symbiotic Plasmid pJB5JI in Nonsterile Soil. AB - Transfer of the pea (Pisum sativum L.) symbiotic plasmid pJB5JI between strains of rhizobia was examined in sterile and nonsterile silt loam soil. Sinorhizobium fredii USDA 201 and HH003 were used as plasmid donors, and symbiotic plasmid cured Rhizobium leguminosarum 6015 was used as the recipient. The plasmid was carried but not expressed in S. fredii strains, whereas transfer of the plasmid to R. leguminosarum 6015 rendered the recipient capable of nodulating pea plants. Confirmation of plasmid transfer was obtained by acquisition of plasmid-encoded antibiotic resistance genes, nodulation of pea plants, and plasmid profiles. Plasmid transfer in nonsterile soil occurred at frequencies of up to 10 per recipient and appeared to be highest at soil temperatures and soil moisture levels optimal for rhizobial growth. Conjugation frequencies were usually higher in sterile soil than in nonsterile soil. In nonsterile soil, transconjugants were recovered only with strain USDA 201 as the plasmid donor. Increasing the inoculum levels of donor and recipient strains up to 10 cells g of soil increased the number of transconjugants; peak plasmid transfer frequencies, however, were found at the lower inoculum level of 10 cells g of soil. Plasmid transfer frequencies were raised in the presence of the pea rhizosphere or by additions of plant material. Transconjugants formed by the USDA 201(pJB5JI) x 6015 mating in soil formed effective nodules on peas. PMID- 16348590 TI - High-Level Nickel Resistance in Alcaligenes xylosoxydans 31A and Alcaligenes eutrophus KTO2. AB - Two new nickel-resistant strains of Alcaligenes species were selected from a large number (about 400) of strains isolated from ecosystems polluted by heavy metals and were studied on the physiological and molecular level. Alcaligenes xylosoxydans 31A is a heterotrophic bacterium, and Alcaligenes eutrophus KTO2 is an autotrophic aerobic hydrogen-oxidizing bacterium. Both strains carry-among other plasmids-a megaplasmid determining resistance to 20 to 50 mM NiCl(2) and 20 mM CoCl(2) (when growing in defined Tris-buffered media). Megaplasmids pTOM8, pTOM9 from strain 31A, and pGOE2 from strain KTO2 confer nickel resistance to the same degree to transconjugants of all strains of A. eutrophus tested but were not transferred to Escherichia coli. However, DNA fragments carrying the nickel resistance genes, cloned into broad-hostrange vector pVDZ'2, confer resistance to A. eutrophus derivatives as well as E. coli. The DNA fragments of both bacteria, TBA8, TBA9, and GBA (14.5-kb BamHI fragments), appear to be identical. They share equal size, restriction maps, and strong DNA homology but are largely different from fragment HKI of nickel-cobalt resistance plasmid pMOL28 of A. eutrophus CH34. PMID- 16348591 TI - Branched-Chain Amino Acid Transport in Cytoplasmic Membranes of Leuconostoc mesenteroides subsp. dextranicum CNRZ 1273. AB - Membrane vesicles of Leuconostoc mesenteroides subsp. dextranicum fused with proteoliposomes prepared from Escherichia coli phospholipids containing beef heart cytochrome c oxidase were used to study the transport of branched-chain amino acids in a strain isolated from a raw milk cheese. At a medium pH of 6.0, oxidation of an electron donor system comprising ascorbate, N,N,N',N'-tetramethyl p-phenylenediamine, and horse heart cytochrome c resulted in a membrane potential (Deltapsi) of -60 mV, a pH gradient of -36 mV, and an l-leucine accumulation of 76-fold (Deltamu(Leu)/F = 108 mV). Leucine uptake in hybrid membranes in which a Deltapsi, DeltapH, sodium ion gradient, or a combination of these was imposed artificially revealed that both components of the proton motive force (Deltap) could drive leucine uptake but that a chemical sodium gradient could not. Kinetic analysis of leucine (valine) transport indicated three secondary transport systems with K(t) values of 1.7 (0.8) mM, 4.3 (5.9) muM, and 65 (29) nM, respectively. l-Leucine transport via the high-affinity leucine transport system (K(t) = 4.3 muM) was competitively inhibited by l-valine and l-isoleucine (K(i) and K(t) values were similar), demonstrating that the transport system translocates branched-chain amino acids. Similar studies with these hybrid membranes indicated the presence of high-affinity secondary transport systems for 10 other amino acids. PMID- 16348592 TI - Study of the Citrate Metabolism of Lactococcus lactis subsp. lactis Biovar Diacetylactis by Means of C Nuclear Magnetic Resonance. AB - The metabolic fate of citrate and pyruvate in four strains of Lactococcus lactis subsp. lactis biovar diacetylactis has been studied by means of C nuclear magnetic resonance, using as a substrate either [3-C]pyruvic acid or custom synthesized citric acid that is C labeled either at carbons 2 and 4 or at carbon 3. The fermentations were carried out batchwise in modified M17 broth. For the actual conversions of the C-labeled substrates, cells at the end of their logarithmic growth phase were used to minimize the conversion to lactic acid. A mass balance of the main citric acid metabolites was obtained; the four strains produced from 50 to 70% (on a molar basis) lactic acid from either citrate or pyruvate. The remaining 50 to 30% was converted mainly to either alpha acetolactic acid (for one strain) or acetoin (for the other three strains). One of the strains produced an exceptionally high concentration of the diacetyl precursor alpha-acetolactic acid. Another strain (SDC6) also produced alpha acetolactic acid, but this was decarboxylated to acetoin at a high rate. The C nuclear magnetic resonance method confirmed that the biosynthesis of alpha acetolactic acid occurs via condensation of pyruvate and "active" acetaldehyde. Diacetyl was not found as a direct metabolite of citrate or pyruvate metabolism. PMID- 16348593 TI - Gibberellins and Carotenoids in the Wild Type and Mutants of Gibberella fujikuroi. AB - A new screening procedure was used to isolate 14 gib mutants of Gibberella fujikuroi with modifications in the production of gibberellins. The production of carotenoids and gibberellins was investigated in the gib mutants and in representative car mutants with various modifications of carotenoid biosynthesis. The determinations of gibberellins were carried out with a simplified fluorescence method. One of the mutants lacked both gibberellins and carotenoids. In many mutants the two pathways compensated each other: an increase in the production of one group of compounds was accompanied by a decrease in the production of the other. Under certain conditions the compensation was quantitative when the output of the two pathways was measured in moles of the common precursor, geranylgeranyl pyrophosphate. alpha-Picoline, an inhibitor of lycopene cyclase in G. fujikuroi, inhibits gibberellin biosynthesis. Other agents that affect the accumulation of carotenoids have no noticeable effect on the accumulation of gibberellins; such is the case with diphenylamine and beta ionone, two inhibitors of phytoene dehydrogenation, and visible light, which stimulates carotenogenesis. PMID- 16348594 TI - Production and Extracellular Secretion of Aqualysin I (a Thermophilic Subtilisin Type Protease) in a Host-Vector System for Thermus thermophilus. AB - Aqualysin I is synthesized as a large precursor, processed, and secreted into the culture medium by Thermus aquaticus YT-1. An expression plasmid for the aqualysin I gene in T. thermophilus HB27 was constructed. T. thermophilus cells harboring the recombinant plasmid produced correctly processed aqualysin I, and the mature enzyme was secreted into the culture medium. PMID- 16348595 TI - Construction of a Bacteriophage-Resistant Derivative of Lactococcus lactis subsp. lactis 425A by Using the Conjugal Plasmid pNP40. AB - Lactococcus lactis subsp. lactis 425A is an atypical strain which excretes a high concentration of alpha-acetolactate when grown in milk. The conjugative lactococcal plasmid pNP40, which encodes phage and nisin resistance, was introduced to strain 425A by conjugation, using resistance to phage and nisin as a selection. No phage-nisin resistance mutants were encountered. Transconjugants display complete resistance at both 21 and 39 degrees C to those phage previously identified as lytic for 425A. Transconjugants lose their resistance characteristics when spontaneously cured of pNP40. The commercially important property of 425A-production of high levels of alpha-acetolactic acid-is unaffected by the presence of pNP40. PMID- 16348596 TI - Screening of natural waters for viruses which infect chlorella cells. AB - By using a plaque assay with the unicellular green alga Chlorella sp. strain NC64A as a host, viruses were screened from natural pond waters collected in Kyoto and Higashi-Hiroshima, Japan. From some samples tested, two kinds of plaques, large (phi = 6 to 10 mm) and small (phi = 2 to 3 mm), were detected with various frequencies. The frequency of plaques in each of the water sources was seasonal; generally, it reached a peak value (8,000 PFU/ml) in May and gradually decreased to the limit of detection (<1) in November before increasing again in early spring. Electron microscopy revealed that the purified and negatively stained viruses were very large (125 to 200 nm) icosahedral particles. The genome isolated from these particles was always a linear double-stranded DNA of 340 to 370 kbp. Electrophoresis patterns of the DNA fragments produced by digestion with restriction enzymes differed considerably from plaque to plaque, even for plaques from the same water source. However, Southern hybridization showed strong homology among all of the virus DNAs tested, indicating relatedness of those viruses. A possible use of the Chlorella virus assay system to monitor the natural population of algal cells and water quality is discussed. PMID- 16348597 TI - Serogrouping of halophilic bdellovibrios from chesapeake bay and environs by immunodiffusion and immunoelectrophoresis. AB - Little has been reported on the serological relationship of halophilic bdellovibrios (Bd). Immunodiffusion analysis performed with rabbit or mouse Bd antisera developed against eight halophilic Bd isolates and one terrestrial Bd isolate, when reacted with soluble antigen preparations of 45 isolates of halophilic Bd, allowed separation into seven serogroups, which were distinct from the terrestrial isolate. Soluble antigen preparations of prey bacteria, Vibrio parahaemolyticus P-5 (P-5) and Escherichia coli ML 35 (ML 35), exhibited no reactivity with the antisera by immunodiffusion. Immunoelectrophoresis revealed the presence of three distinct antigens in homologous reactions and one shared antigen in heterologous Bd reactions. Shared antigens were noted between halophilic and terrestrial Bd, in addition to between halophilic Bd strains, indicating the possible existence of an antigen(s) which may be shared among all Bd. Again, no shared antigen was noted when P-5 or ML 35 was allowed by immunoelectrophoresis to react with the antisera. Prey susceptibility testing of the seven distinct groups of halophilic Bd, using 20 test prey, produced essentially identical spectra for each group, indicating that this was not a useful technique in delineating the Bd. While immunoelectrophoresis was able to demonstrate an antigen common to all Bd tested, immunodiffusion was able to delineate strains on the basis of a "serogroup specific" antigen. This suggests that immunological tools may serve as important means to study the taxonomy of halophilic Bd, as well as in the formation of a clearer taxonomic picture of the genus Bdellovibrio. PMID- 16348598 TI - Measurement of acetate concentrations in marine pore waters by using an enzymatic approach. AB - Acetate concentrations in marine and freshwater matrices were measured by an enzymatic technique which coupled the synthesis of acetyl coenzyme A to AMP production. The resulting AMP was assayed by a sensitive and relatively rapid high-pressure liquid chromatography method, using an aqueous, isocratic mobile phase for elution. The method was insensitive to the presence of seawater salts and required no sample prepurification or distillation. Propionate caused a minor, but statistically insignificant, interference when equimolar with acetate; butyrate caused no interference, even at relatively high concentrations. Detection limits for acetate were approximately 100 nM with a precision of about 5%. Pore waters from two intertidal sediments contained approximately 1 to 12 muM acetate; the concentrations were linearly but inversely correlated with porewater sulfate. PMID- 16348599 TI - Detection of Plasmid Transfer from Pseudomonas fluorescens to Indigenous Bacteria in Soil by Using Bacteriophage phiR2f for Donor Counterselection. AB - The transfer of a genetically marked derivative of plasmid RP4, RP4p, from Pseudomonas fluorescens to members of the indigenous microflora of the wheat rhizosphere was studied by using a bacteriophage that specifically lyses the donor strain and a specific eukaryotic marker on the plasmid. Transfer of RP4p to the wheat rhizosphere microflora was observed, and the number of transconjugants detected was approximately 10 transconjugants per g of soil when 10 donor cells per g of soil were added; transfer in the corresponding bulk soil was slightly above the limit of detection. All of the indigenous transconjugants which we analyzed contained a 60-kb plasmid and were able to transfer this plasmid to a Nx RpP. fluorescens recipient strain. The indigenous transconjugants were identified as belonging to Pseudomonas spp., Enterobacter spp., Comamonas spp., and Alcaligenes spp. PMID- 16348600 TI - Genetic Diversity among Rhizobium leguminosarum bv. Trifolii Strains Revealed by Allozyme and Restriction Fragment Length Polymorphism Analyses. AB - Allozyme electrophoresis and restriction fragment length polymorphism (RFLP) analyses were used to examine the genetic diversity of a collection of 18 Rhizobium leguminosarum bv. trifolii, 1 R. leguminosarum bv. viciae, and 2 R. meliloti strains. Allozyme analysis at 28 loci revealed 16 electrophoretic types. The mean genetic distance between electrophoretic types of R. leguminosarum and R. meliloti was 0.83. Within R. leguminosarum, the single strain of bv. viciae differed at an average of 0.65 from strains of bv. trifolii, while electrophoretic types of bv. trifolii differed at a range of 0.23 to 0.62. Analysis of RFLPs around two chromosomal DNA probes also delineated 16 unique RFLP patterns and yielded genetic diversity similar to that revealed by the allozyme data. Analysis of RFLPs around three Sym (symbiotic) plasmid-derived probes demonstrated that the Sym plasmids reflect genetic divergence similar to that of their bacterial hosts. The large genetic distances between many strains precluded reliable estimates of their genetic relationships. PMID- 16348601 TI - Isolation and Characterization of a Competition-Defective Bradyrhizobium japonicum Mutant. AB - Tn5 mutagenesis was coupled with a competition assay to isolate mutants of Bradyrhizobium japonicum defective in competitive nodulation. A double selection procedure was used, screening first for altered extracellular polysaccharide production (nonmucoid colony morphology) and then for decreased competitive ability. One mutant, which was examined in detail, was deficient in acidic polysaccharide and lipopolysaccharide production. The wild-type DNA region corresponding to the Tn5 insertion was isolated, mapped, and cloned. A 3.6-kb region, not identified previously as functioning in symbiosis, contained the gene(s) necessary for complementation of the mutation. The mutant was motile, grew normally on minimal medium, and formed nodules on soybean plants which fixed almost as much nitrogen as the wild type during symbiosis. PMID- 16348602 TI - Citrate Fermentation by Lactococcus and Leuconostoc spp. AB - Citrate and lactose fermentation are subject to the same metabolic regulation. In both processes, pyruvate is the key intermediate. Lactococcus lactis subsp. lactis biovar diacetylactis homofermentatively converted pyruvate to lactate at high dilution (growth) rates, low pH, and high lactose concentrations. Mixed-acid fermentation with formate, ethanol, and acetate as products was observed under conditions of lactose limitation in continuous culture at pH values above 6.0. An acetoin/butanediol fermentation with alpha-acetolactate as an intermediate was found upon mild aeration in continuous culture and under conditions of excess pyruvate production from citrate. Leuconostoc spp. showed a limited metabolic flexibility. A typical heterofermentative conversion of lactose was observed under all conditions in both continuous and batch cultures. The pyruvate produced from either lactose or citrate was converted to d-lactate. Citrate utilization was pH dependent in both L. lactis and Leuconostoc spp., with maximum rates observed between pH 5.5 and 6.0. The maximum specific growth rate was slightly stimulated by citrate, in L. lactis and greatly stimulated by citrate in Leuconostoc spp., and the conversion of citrate resulted in increased growth yields on lactose for both L. lactis and Leuconostoc spp. This indicates that energy is conserved during the metabolism of citrate. PMID- 16348603 TI - alpha-Glucuronidase and Other Hemicellulase Activities of Fibrobacter succinogenes S85 Grown on Crystalline Cellulose or Ball-Milled Barley Straw. AB - Fibrobacter succinogenes produces an alpha-glucuronidase which cleaves 4-O-methyl alpha-d-glucuronic acid from birch wood 4-O-methyl-alpha-d-glucuronoxylan. Very low levels of alpha-glucuronidase activity were detected in extracellular enzyme preparations of F. succinogenes on birch wood xylan substrate. The release of 4-O methyl-alpha-d-glucuronic acid was enhanced when the birch wood xylan substrate was predigested by either a purified Schizophyllum commune xylanase or a cloned F. succinogenes S85 xylanase. These data suggest that the alpha-glucuronidase is unable to cleave 4-O-methyl-alpha-d-glucuronic acid from intact xylan but can act on unique low-molecular-weight glucuronoxylan fragments created by the cloned F. succinogenes xylanase. The cloned xylanase presumably must account for a small proportion of the indigenous xylanase activity of F. succinogenes cultures, since this xylanase source does not support high glucuronidase activity. The alpha glucuronidase and associated hemicellulolytic enzymes exhibited higher activities in culture fluid from cells grown on ball-milled barley straw than in that of cellulose-grown cells. The profile of xylanases separated by isoelectric focusing (zymogram) of culture filtrate from cells grown on barley straw was more complex than that of culture filtrates from cells grown on cellulose. These data demonstrate that F. succinogenes produces an alpha-glucuronidase with an exacting substrate specificity which enables extensive cleavage of glucuronic acid residues from xylan as a consequence of synergistic xylanase action. PMID- 16348604 TI - Genetic Interrelatedness among Clover Proliferation Mycoplasmalike Organisms (MLOs) and Other MLOs Investigated by Nucleic Acid Hybridization and Restriction Fragment Length Polymorphism Analyses. AB - DNA was isolated from clover proliferation (CP) mycoplasmalike organism (MLO) diseased periwinkle plants (Catharanthus roseus (L.) G. Don.) and cloned into pSP6 plasmid vectors. CP MLO-specific recombinant DNA clones were biotin labeled and used as probes in dot hybridization and restriction fragment length polymorphism analyses to study the genetic interrelatedness among CP MLO and other MLOs, including potato witches'-broom (PWB) MLO. Results from dot hybridization analyses indicated that both a Maryland strain of aster yellows and a California strain of aster yellows are distantly related to CP MLO. Elm yellows, paulownia witches'-broom, peanut witches'-broom, loofah witches'-broom, and sweet potato witches'-broom may be very distantly related, if at all, to CP MLO. A new Jersey strain of aster yellows MLO, tomato big bud MLO, clover phyllody MLO, beet leafhopper-transmitted virescence MLO, and ash yellows MLO are related to CP MLO, but PWB MLO is the most closely related. Similarity coefficients derived from restriction fragment length polymorphism analyses revealed that PWB and CP MLOs are closely related strains and thus provided direct evidence of their relatedness in contrast to reliance solely on biological characterization. PMID- 16348605 TI - Cloning and detection of chromosomal and extrachromosomal DNA from mycoplasmalike organisms that cause yellow dwarf disease of rice. AB - DNA was extracted from rice plants infected with mycoplasmalike organisms (MLOs) causing yellow dwarf disease. DNA of the causal agent was separated from the host DNA by repeated bisbenzimide-CsCl equilibrium density gradient centrifugations. MLO DNA cut by HindIII was ligated into plasmid Bluescript II and cloned in Escherichia coli NM522. The DNA inserts were labeled with peroxidase and employed as probes in hybridization. Southern analysis revealed that the insert in pRYD-12 consisted of one, presumably chromosomal, piece of MLO DNA, whereas the insert in pRYD-19, another recombinant plasmid, consisted of one, presumably extrachromosomal, piece of MLO DNA. Cloned DNA probes were successfully applied in dot blot hybridization for the detection of rice yellow dwarf disease MLOs in rice plants and in an insect vector, the green rice leafhopper (Nephotettix cincticeps). PMID- 16348606 TI - Effects of hyperbaric pressure on a deep-sea archaebacterium in stainless steel and glass-lined vessels. AB - The effects of hyperbaric helium pressures on the growth and metabolism of the deep-sea isolate ES4 were investigated. In a stainless steel reactor, cell growth was completely inhibited but metabolic gas production was observed. From 85 to 100 degrees C, CO(2) production proceeded two to three times faster at 500 atm (1 atm = 101.29 kPa) than at 8 atm. At 105 degrees C, no CO(2) was produced until the pressure was increased to 500 atm. Hydrogen and H(2)S were also produced biotically but were not quantifiable at pressures above 8 atm because of the high concentration of helium. In a glass-lined vessel, growth occurred but the growth rate was not accelerated by pressure. In most cases at temperatures below 100 degrees C, the growth rate was lower at elevated pressures; at 100 degrees C, the growth rates at 8, 250, and 500 atm were nearly identical. Unlike in the stainless steel vessel, CO(2) production was exponential during growth and continued for only a short time after growth. In addition, relatively little H(2) was produced in the glass-lined vessel, and there was no growth or gas production at 105 degrees C at any pressure. The behavior of ES4 as a function of temperature and pressure was thus very sensitive to the experimental conditions. PMID- 16348607 TI - New routes for aerobic biodegradation of dimethylsulfoniopropionate. AB - Dimethylsulfoniopropionate (DMSP), an osmolyte in marine plants, is biodegraded by cleavage of dimethyl sulfide (DMS) or by demethylation to 3-methiolpropionate (MMPA) and 3-mercaptopropionate (MPA). Sequential demethylation has been observed only with anoxic slurries of coastal sediments. Bacteria that grew aerobically on MMPA and DMSP were isolated from marine environments and phytoplankton cultures. Enrichments with DMSP selected for bacteria that generated DMS, whereas MMPA enrichments selected organisms that produced methanethiol (CH(3)SH) from either DMSP or MMPA. A bacterium isolated on MMPA grew on MMPA and DMSP, but rapid production of CH(3)SH from DMSP occurred only with DMSP-grown cells. Low levels of MPA accumulated during growth on MMPA, indicating demethylation as well as demethiolation of MMPA. The alternative routes for DMSP biodegradation via MMPA probably impact on net DMS fluxes to the marine atmosphere. PMID- 16348608 TI - Nitrification at Low pH by Aggregated Chemolithotrophic Bacteria. AB - A study was performed to gain insight into the mechanism of acid-tolerant, chemolithotrophic nitrification. Microorganisms that nitrified at pH 4 were enriched from two Dutch acid soils. Nitrate production in the enrichment cultures was indicated to be of a chemolithoautotrophic nature as it was (i) completely inhibited by acetylene at a concentration as low as 1 mumol/liter and (ii) strongly retarded under conditions of carbon dioxide limitation. Electron microscopy of the enrichment cultures showed the presence of bacteria that were morphologically similar to strains of known chemolithotrophic nitrifying genera. Many of the enriched bacteria, in particular those that were identified as ammonium oxidizers, were aggregated. Filtration experiments indicated that aggregated cells were able to nitrify at low pH, whereas single cells were not. It is hypothesized that cells inside the aggregates are protected against the toxicity of nitrous acid. Nitrification by aggregated chemolithoautotrophic bacteria may be the dominating process of nitrate formation in many acid soils as it does not appear to depend on the existence of microsites of high pH (acid sensitive autotrophic nitrification) or on the availability of organic carbon (heterotrophic nitrification). PMID- 16348609 TI - F Nuclear Magnetic Resonance Analysis of 5-Fluorouracil Metabolism in Four Differently Pigmented Strains of Nectria haematococca. AB - F nuclear magnetic resonance spectroscopy was used to study the metabolism of 5 fluorouracil in four strains of Nectria haematococca which displayed similar sensitivities to growth inhibition by this compound but differed in their pigmentation. The major metabolites, 5-fluorouridine and alpha-fluoro-beta alanine, were excreted into the medium by all four strains. The classical ribofluoronucleotides (5-fluorouridine-5'-monophosphate, -diphosphate, and triphosphate) and alpha-fluoro-beta-alanine were identified in the acid-soluble fraction of perchloric acid extracts of mycelia. Two hydrolysis products of 5 fluorouracil incorporated into RNA were found in the acid-insoluble pool. They were unambiguously assigned to 5-fluorouridine-2'-monophosphate and 3' monophosphate with specific hydrolysis reactions on isolated RNA. The lack of fluorodeoxyribonucleotides and the fact that the four strains incorporated similar amounts of fluororibonucleotides into their RNAs strongly suggest an RNA directed mechanism of cytotoxicity for 5-fluorouracil. The heavily pigmented wild type differed from the three low-pigmented strains in its low uptake of 5 fluorouracil and, consequently, in its reduced biosynthesis of 5-fluorouridine and alpha-fluoro-beta-alanine. At present, it is not clear whether this change in 5-fluorouracil metabolism is a side effect of pigment production or results from another event. PMID- 16348610 TI - Use of Bioluminescence Markers To Detect Pseudomonas spp. in the Rhizosphere. AB - The use of bioluminescence as a sensitive marker for detection of Pseudomonas spp. in the rhizosphere was investigated. Continuous expression of the luxCDABE genes, required for bioluminescence, was not detectable in the rhizosphere. However, when either a naphthalene-inducible luxCDABE construct or a constitutive luxAB construct (coding only for the luciferase) was introduced into the Pseudomonas cells, light emission could be initiated just prior to measurement by the addition of naphthalene or the substrate for luciferase, n-decyl aldehyde, respectively. These Pseudomonas cells could successfully be detected in the rhizosphere by using autophotography or optical fiber light measurement techniques. Detection required the presence of 10 to 10 CFU/cm of root, showing that the bioluminescence technique is at least 1,000-fold more sensitive than beta-galactosidase-based systems. PMID- 16348611 TI - Diglyceride Kinase Activity in Cell Extracts of Rhizobium meliloti: Evidence for a Diglyceride Cycle during Cyclic beta-1,2-Glucan Biosynthesis. AB - In this article, we provide evidence for the presence of diglyceride kinase activity in cell extracts of Rhizobium meliloti 1021. Characterization of the rhizobial enzyme revealed that it shares many properties with the diglyceride kinase of Escherichia coli. A possible role for this enzyme during cyclic beta 1,2-glucan biosynthesis is discussed. PMID- 16348612 TI - Separation of methanotrophic bacteria by using percoll and its application to isolation of mixed and pure cultures. AB - Type I and II methanotrophic bacteria can be cleanly separated by using Percoll buoyant density gradients. This simple and inexpensive method can be used to screen new isolates rapidly for preliminary classification. In addition, Percoll gradients can also be used to enhance standard enrichment techniques for methanotrophs from natural water samples. PMID- 16348613 TI - Improved technique for identification and enumeration of methanogenic bacterial colonies on roll tubes by epifluorescence microscopy. AB - Methanogenic fluorescent colonies can be clearly identified on roll tubes by using an epifluorescence microscope equipped with a x 2 objective. Methanogenic and nonmethanogenic colonies could be counted in roll tubes prepared from methanogenic enrichment cultures. Late-developing colonies appearing after 25 days of incubation were mainly methanogenic. PMID- 16348615 TI - Osmotically Regulated Transport of Proline by Lactobacillus acidophilus IFO 3532. AB - [This corrects the article on p. 2831 in vol. 57.]. PMID- 16348614 TI - New culture medium containing ionic concentrations of nutrients similar to concentrations found in the soil solution. AB - A new growth medium which closely approximates the composition of the soil solution is presented. This soil solution equivalent (SSE) medium contains the following components (millimolar): NO(3), 2.5; NH(4), 2.5; HPO(4), 0.005; Na, 2.5; Ca, 4.0; Mg, 2.0; K, 0.503; Cl, 4.0; SO(4), 5.0; ethylenediamine-di(o hydroxyphenylacetic acid), 0.02; and MES [2-(N-morpholino)ethanesulfonic acid] (to maintain the pH at 6.0), 10, plus 0.1% arabinose. The advantages of the SSE medium are discussed. PMID- 16348616 TI - Treating soil solution samplers to prevent microbial removal of analytes. AB - Soil microorganisms colonizing soil water sampling devices (lysimeters) reduced concentrations of biodegradable organic chemicals, including 2,4 dichlorophenoxyacetic acid methyl ester, alachlor, methyl m-chlorobenzoate, and metolachlor as water entered through porous ceramic cups. In some cases, losses exceeded 99%. Additions of either a biocide (sodium hypochlorite) or a bacteriostat (copper salt) prevented microbial activity so that concentrations of test chemicals inside lysimeters equaled those outside. Field studies further indicated that treating lysimeters with a copper salt effectively prevented microbial activity. Thus, chemically treating soil water samplers could improve the accuracy of soil water data for a wide variety of analytes, including environmentally important organics, such as pesticides and industrial wastes, and inorganics, such as ammonia and nitrate. PMID- 16348617 TI - Induction of Cellulase by Gentiobiose and Its Sulfur-Containing Analog in Penicillium purpurogenum. AB - Cellulase induction by beta-glucodisaccharides was investigated by using non cellulase-induced mycelia of Penicillium purpurogenum P-26, a highly-cellulase producing fungus. Gentiobiose induced significant amounts of cellulase compared with cellobiose when nojirimycin was added to the induction medium to inhibit extracellular beta-glucosidase activity. Thiogentiobiose (6-S-beta-d glucopyranosyl-6-thio-d-glucose), a sulfur-containing analog of gentiobiose, was more effective for cellulase induction than gentiobiose even in the absence of nojirimycin. Thiogentiobiose appeared to be a gratuitous inducer since it was not metabolized during cellulase induction. Gentiobiose was formed from cellobiose by the intracellular beta-glucosidase of P. purpurogenum. These findings indicate that gentiobiose is an active inducer of cellulase for this fungus and may possibly be formed by intracellular beta-glucosidase from cellobiose. PMID- 16348618 TI - Differential siderophore utilization and iron uptake by soil and rhizosphere bacteria. AB - The differential availabilities of the hydroxamate siderophores ferrioxamine B (FOB) and ferrichrome (FC) and the pseudobactin siderophores St3, 7NSK(2), and WCS 358 as sources of Fe for soil and rhizosphere bacteria were studied. About 20% of the total bacterial CFU from the rhizospheres of four plant species were able to use FOB as the sole Fe source in an Fe-deficient medium, while about 12, 10, 2, and > 1% were able to use FC and pseudobactins 7NSK(2), St3, and WCS 358, respectively. Of the 165 colonies isolated from plates containing pseudobactins, 64 were able to use the pseudobactin on which they were isolated as the sole Fe source in pure culture. Cross-feeding tests showed that almost all of these 64 strains were also able to use at least one of the other siderophores studied (pseudobactin, FOB, or FC). Pseudomonas putida StS2, Pseudomonas maltophilia 7NM1, and Vibrio fluvialis WS1, which were originally isolated on pseudobactins St3, 7NSK(2), and WCS 358, respectively, were selected for their ability to grow with pseudobactin St3 as the sole Fe source. They incorporated Fe mediated by pseudobactin St3 at various rates (71.5, 4, and 23 pmol/min/mg [dry weight] of cells, respectively). Similarly, P. putida St3 was shown to incorporate Fe mediated by FOB and FC. We suggest that the ability of bacteria to utilize a large variety of siderophores confers an ecological advantage. PMID- 16348620 TI - Effects of nutrients on specific growth rate of bacterioplankton in oligotrophic lake water cultures. AB - The effects of organic and inorganic nutrient additions on the specific growth rates of bacterioplankton in oligotrophic lake water cultures were investigated. Lake water was first passed through 0.8-mum-pore-size filters (prescreening) to remove bacterivores and to minimize confounding effects of algae. Specific growth rates were calculated from changes in both bacterial cell numbers and biovolumes over 36 h. Gross specific growth rates in unmanipulated control samples were estimated through separate measurements of grazing losses by use of penicillin. The addition of mixed organic substrates alone to prescreened water did not significantly increase bacterioplankton specific growth rates. The addition of inorganic phosphorus alone significantly increased one or both specific growth rates in three of four experiments, and one experiment showed a secondary stimulation by organic substrates. The stimulatory effects of phosphorus addition were greatest concurrently with the highest alkaline phosphatase activity in the lake water. Because bacteria have been shown to dominate inorganic phosphorus uptake in other P-deficient systems, the demonstration that phosphorus, rather than organic carbon, can limit bacterioplankton growth suggests direct competition between phytoplankton and bacterioplankton for inorganic phosphorus. PMID- 16348619 TI - Production of an Amylase-Sensitive Bacteriocin by an Atypical Leuconostoc paramesenteroides Strain. AB - An atypical Leuconostoc paramesenteroides strain isolated from retail lamb produced a bacteriocin, leuconocin S, that was inactivated by alpha-amylase, trypsin, alpha-chymotrypsin, protease, and proteinase K but not by lipase or heat treatment at 60 degrees C for 30 min. Supernatants from culture broths produced two glycoprotein bands on sodium dodecyl sulfate-polyacrylamide gels; these had molecular weights of 2,000 and 10,000 and activity against Lactobacillus sake ATCC 15521. The crude bacteriocin preparation was bacteriostatic and dissipated proton motive force. Bacteriocin activity was produced over a wide pH range (5.2 to 7.9) on buffered agar medium, with an optimum pH of pH 6.15. The optimum pH for production in broth was 6.5 to 7.0. PMID- 16348621 TI - High-Frequency Plasmid Transduction by Lactobacillus gasseri Bacteriophage phiadh. AB - The temperate bacteriophage phiadh mediates plasmid DNA transduction in Lactobacillus gasseri ADH at frequencies in the range of 10 to 10 transductants per PFU. BglII-generated DNA fragments from phage phiadh were cloned into the BclI site of the transducible plasmid vector pGK12 (4.4 kb). Phage phiadh lysates induced from Lactobacillus lysogens harboring pGK12 or the recombinant plasmids were used to transduce strain ADH to chloramphenicol resistance. The transduction frequencies of recombinant plasmids were 10- to 10-fold higher than that of native pGK12. The increase in frequency generally correlated with the extent of DNA-DNA homology between plasmid and phage DNAs. The highest transduction frequency was obtained with plasmid pTRK170 (6.6 kb), a pGK12 derivative containing the 1.4- and 0.8-kb BglII DNA fragments of phiadh. DNA hybridization analysis of pTRK170-transducing phage particles revealed that pTRK170 had integrated into the phiadh genome, suggesting that recombination between homologous sequences present in phage and plasmid DNAs was responsible for the formation of high-frequency transducing phage particles. Plasmid DNA analysis of 13 transductants containing pTRK170 showed that each had acquired intact plasmids, indicating that in the process of transduction a further recombination step was involved in the resolution of plasmid DNA monomers from the recombinant pTRK170::phiadh molecule. In addition to strain ADH, pTRK170 could be transduced via phiadh to eight different L. gasseri strains, including the neotype strain, F. Gasser 63 AM (ATCC 33323). PMID- 16348622 TI - Sheared-root inocula of vesicular-arbuscular mycorrhizal fungi. AB - For efficient handling, vesicular-arbuscular mycorrhizal fungi should be processed into small and uniform inocula; however, processing can reduce the inoculum density. In this article we describe the preparation and use of sheared root inocula of Glomus spp. in which inoculum densities were increased during processing. Our objectives were to determine inoculum viability and density after shearing and to ascertain if the sheared inocula could be pelletized or used with a gel carrier. Root samples were harvested from aeroponic cultures, blotted dry, cut into 1-cm lengths, and sheared in a food processor for up to 80 s. After shearing, the inoculum was washed over sieves, and the propagule density in each fraction was determined. Sheared inocula were also encapsulated in carrageenan or used in a gel carrier. Shearing aeroponically produced root inocula reduced particle size. Propagule density increased with decreasing size fraction down to a size of 63 mum, after which propagule density decreased. The weighted-average propagule density of the inoculum was 135,380 propagules g (dry weight) of sheared root material. Sheared roots were encapsulated successfully in carrageenan, and the gel served as an effective carrier. Aeroponic root inoculum was stored dry at 4 degrees C for 23 months without significant reduction in propagule density; however, this material was not appropriate for shearing. Moist roots, useful for shearing, began to lose propagule density after 1 month of storage. Shearing proved to be an excellent method to prepare viable root inocula of small and uniform size, allowing for more efficient and effective use of limited inoculum supplies. PMID- 16348623 TI - Fate of Fumonisin B(1) in Naturally Contaminated Corn during Ethanol Fermentation. AB - Two lots of corn naturally contaminated with fumonisin B(1) (15 and 36 ppm) and a control lot (no fumonisin B(1) detected) were used as substrates for ethanol production in replicate 8.5-liter yeast fermentations. Ethanol yields were 8.8% for both the control and low-fumonisin corn, while the high-fumonisin corn contained less starch and produced 7.2% ethanol. Little degradation of fumonisin occurred during fermentation, and most was recovered in the distillers' grains, thin stillage, and distillers' solubles fractions. No toxin was detected in the distilled alcohol or centrifuge solids. Ethanol fermentation of fumonisin contaminated corn coupled with effective detoxification of distillers' grains and aqueous stillage is suggested as a practical process strategy for salvaging contaminated corn. PMID- 16348624 TI - Regulatory influences on the production of gamma-aminobutyric Acid by a marine pseudomonad. AB - A pseudomonad capable of producing gamma-aminobutyric acid (GABA) was isolated from seawater via an enrichment in which glutamate was the sole carbon and nitrogen source. The organism grew optimally at pH 7.3 and at 25 degrees C. Putrescine, alanine, and glucose-nitrate also served as effective growth substrates. The isolate grew poorly on GABA. Cell suspensions of the organism in 0.02 M phosphate buffer (pH 7.6) containing NaCl (19.4 g liter) and MgCl(2). 6H(2)O(3 g liter) produced GABA from succinic semialdehyde in combination with glutamate or alanine but not from any substrate alone. Little or no GABA was produced with putrescine or glucose-nitrate as substrates. GABA production in the amino acid cosubstrate systems was transitory with optimum levels occurring in the suspension fluid after 3 h of incubation (0.3 and 0.03 mM for glutamate and alanine cosubstrates, respectively). However, yields of GABA in the cell suspension fluid were low, and quantities near that predicted from stoichiometry could be obtained only by extracting cell suspensions with methanol. GABA release in the suspension fluid was increased with higher pH or by decreasing NaCl. Substitution of the salt by the equivalent Tris-HCl or KCl likewise resulted in increased GABA release. When nigericin (10 mug ml) was added to cell suspensions in which NaCl was not decreased, GABA release increased in a way similar to that observed in suspensions with decreased NaCl. The ionophore also decreased GABA uptake by cell suspensions of GABA-grown cells, and the effect was duplicated by lowering NaCl in cell suspensions. The results indicate a role for an Na dependent transport system in GABA release. PMID- 16348626 TI - Isolation and Characterization of Haloanaerobacter chitinovorans gen. nov., sp. nov., a Halophilic, Anaerobic, Chitinolytic Bacterium from a Solar Saltern. AB - Two halophilic anaerobic bacteria, one of which had chitinolytic activity, were isolated from a solar saltern in southern California. These organisms were long, gram-negative, motile, flexible rods. The biochemical and physiological characteristics of these bacteria were very similar but were different from the characteristics of other haloanaerobic bacteria. Both grew at salt concentrations ranging from 0.5 to 5 M and at temperatures ranging from 23 to 50 degrees C. They were sensitive to chloramphenicol but resistant to penicillin, carbenicillin, d cycloserine, streptomycin, and tetracycline. An analysis of DNAs and whole-cell proteins showed that they were closely related taxonomically and distinguishable from other halophilic anaerobic bacteria. They exhibited 92.3 to 100% DNA homology as determined by DNA-DNA hybridization. The guanine-plus-cytosine contents of their DNAs were 34.8+/-1 mol%. The two isolates, strains W5C8 and W3C1, differed from other halophilic anaerobic bacteria sufficiently to support establishment of a new genus and species, Haloanaerobacter chitinovorans. Strain W5C8 exhibited chitinolytic activity and is designated the type strain. Two chitin-induced extracellular proteins with molecular weights of 38 x 10 and 40 x 10 were detected in strain W5C8. PMID- 16348625 TI - Survival of Bacillus licheniformis in Seawater Model Ecosystems. AB - The fate of Bacillus licheniformis DSM 13 was monitored after introduction into laboratory microcosms and mesocosms established in the Knebel Vig estuary, Denmark. The model organism was detected by a combination of immunofluorescence microscopy and nonselective plating followed by colony blotting. This allowed simultaneous quantification of intact cells and culturable cells. B. licheniformis DSM 13 adapted poorly to the conditions in filtered (0.2-mum-pore size filter) seawater. Results from additional microcosm studies using natural seawater demonstrated that protozoan grazing also was important in regulating the population of the introduced model organism. In experiments using mesocosms, B. licheniformis DSM 13 also showed a rapid die-off. The introduction of the organism led to increased nutrient levels and to increased growth of both autotrophic and heterotrophic components of the plankton community compared with those of control enclosures. Thereby, a more intensive predation impact on the bacterioplankton community was induced. The combination of microcosm and mesocosm experiments provides a scenario in which the influence of single biotic and abiotic factors on survival of introduced organisms can be tested and in which the effect of the introduction on ecosystem structure and function can be evaluated. This test concept might prove useful in risk assessment of genetically modified microorganisms. PMID- 16348627 TI - Accumulation of copper and other metals by copper-resistant plant-pathogenic and saprophytic pseudomonads. AB - Copper-resistant strains of Pseudomonas syringae carrying the cop operon produce periplasmic copper-binding proteins, and this sequestration outside the cytoplasm has been proposed as a resistance mechanism. In this study, strain PS61 of P. syringae carrying the cloned cop operon accumulated more total cellular copper than without the operon. Several other copper-resistant pseudomonads with homology to cop were isolated from plants, and these bacteria also accumulated copper. Two highly resistant species accumulated up to 115 to 120 mg of copper per g (dry weight) of cells. P. putida 08891 was more resistant to several metals than P. syringae pv. tomato PT23, but this increased resistance was not correlated with an increased accumulation of metals other than copper. Several metals were accumulated by both PT23 and P. putida, but when copper was added to induce the cop operon, there was generally no increase of accumulation of the other metals, suggesting that the cop operon does not contribute to accumulation of these other metals. The exceptions were aluminum for PT23 and iron for P. putida, which accumulated to higher levels when copper was added to the cultures. The results of this study support the role of copper sequestration in the copper resistance mechanism of P. syringae and suggest that this mechanism is common to several copper-resistant Pseudomonas species found on plants to which antimicrobial copper compounds are applied for plant disease control. PMID- 16348628 TI - Purification and Partial Characterization of Lacticin 481, a Lanthionine Containing Bacteriocin Produced by Lactococcus lactis subsp. lactis CNRZ 481. AB - Lacticin 481, a bacteriocin produced during the growth of Lactococcus lactis subsp. lactis CNRZ 481, was purified sequentially by ammonium sulfate precipitation, gel filtration, and preparative and analytical reversed-phase high pressure liquid chromatography. Ammonium sulfate precipitations resulted in a 455 fold increase in total lacticin 481 activity. The entire purification protocol led to a 107, 506-fold increase in the specific activity of lacticin 481. On the basis of its electrophoretic pattern in sodium dodecyl sulfate-polyacrylamide gels, lacticin 481 appeared as a single peptide band of 1.7 kDa. However, dimers of 3.4 kDa also exhibiting lacticin activity were detected. Derivatives of the lacticin-producing strain which did not produce lacticin 481 (Bac) were sensitive to this bacteriocin (Bac) and failed to produce the 1.7-kDa band. Amino acid composition analysis of purified lacticin 481 revealed the presence of lanthionine residues, suggesting that lacticin 481 is a member of the lantibiotic family of antimicrobial peptides. Seven residues (K G G S G V I) were sequenced from the N-terminal portion of lacticin 481, and these did not shown any homology with nisin or other known bacteriocin sequences. PMID- 16348629 TI - Localization of peptidases in lactococci. AB - The localization of two aminopeptidases, an X-prolyl-dipeptidyl aminopeptidase, an endopeptidase, and a tripeptidase in Lactococcus lactis was studied. Polyclonal antibodies raised against each purified peptidase are specific and do not cross-react with other peptidases. Experiments were performed by immunoblotting after cell fractionation and by electron microscopy of immunogold labeled peptidases. All peptidases were found to be intracellular. However, immunogold studies showed a peripheral labeling of the X-prolyl-dipeptidyl aminopeptidase, the tripeptidase, and the endopeptidase. This peripheral location was further supported by the detection of these three enzymes in cell membrane fractions in which none of the two aminopeptidases was present. PMID- 16348630 TI - Denitrification, dissimilatory reduction of nitrate to ammonium, and nitrification in a bioturbated estuarine sediment as measured with N and microsensor techniques. AB - Nitrogen and oxygen transformations were studied in a bioturbated (reworked by animals) estuarine sediment (Norsminde Fjord, Denmark) by using a combination of N isotope (NO(3)), specific inhibitor (C(2)H(2)), and microsensor (N(2)O and O(2)) techniques in a continuous-flow core system. The estuarine water was NO(3) rich (125 to 600 muM), and NO(3) was consistently taken up by the sediment on the four occasions studied. Total NO(3) uptake (3.6 to 34.0 mmol of N m day) corresponded closely to N(2) production (denitrification) during the experimental steady state, which indicated that dissimilatory, as well as assimilatory, NO(3) reduction to NH(4) was insignificant. When C(2)H(2) was applied in the flow system, denitrification measured as N(2)O production was often less (58 to 100%) than the NO(3) uptake because of incomplete inhibition of N(2)O reduction. The NO(3) formed by nitrification and not immediately denitrified but released to the overlying water, uncoupled nitrification, was calculated both from NO(3) dilution and from changes in NO(3) uptake before and after C(2)H(2) addition. These two approaches gave similar results, with rates ranging between 0 and 8.1 mmol of N m day on the four occasions. Attempts to measure total nitrification activity by the difference between NH(4) fluxes before and after C(2)H(2) addition failed because of non-steady-state NH(4) fluxes. The vertical distribution of denitrification and oxygen consumption was studied by use of N(2)O and O(2) microelectrodes. The N(2)O profiles measured during the experimental steady state were often irregularly shaped, and the buildup of N(2)O after C(2)H(2) was added was much too fast to be described by a simple diffusion model. Only bioturbation by a dense population of infauna could explain these observations. This was corroborated by the relationship between diffusive and total fluxes, which showed that only 19 to 36 and 29 to 62% of the total O(2) uptake and denitrification, respectively, were due to diffusion-reaction processes at the regular sediment surface, excluding animal burrows. PMID- 16348631 TI - Competitive ability and survival in soil of pseudomonas strain 679-2, a dominant, nonobligate bacterial predator of bacteria. AB - A copper-resistant, nonobligate, bacterial predator of bacteria was isolated from soil. It was a Pseudomonas species, designated strain 679-2. It attacked most other nonobligate bacterial predators and hence could control their predatory and other activities in nature. It also inhibited various fungi. It attached to prey cells and produced a toxic, copper-related, growth initiation factor like that produced by Cupriavidus necator. In addition, it produced a second, novel compound that was both antibacterial and antifungal. Strain 679-2 appeared to have only a very limited natural occurrence. It was found only in the soil from one small area in one field. It was absent on the leaves of the plant species that were examined. Regardless of its rarity, however, it was highly competitive in soil. An inoculum consisting of only a few cells added to soil multiplied rapidly to become a major component of the soil microflora within 24 h. A small amount of glutamic acid could be added along with the cells to stimulate production of the toxic compounds noted above, but this was not necessary. After this multiplication, or when large numbers of cells were added to soil, the numbers decreased only slowly during the next several months. Cell survival also was good on plant leaves. The survival in soil and on plant leaves occurred in both laboratory and field experiments. Other than desiccation, the natural mechanism for controlling the numbers or activities of strain 679-2 in soil is not known. The various characteristics of this bacterium, as noted above, are of particular interest because they indicate a possible use of the cells or inhibitor compounds for controlling organisms in soil or on plant surfaces. PMID- 16348632 TI - Enrichment of Thermophilic Propionate-Oxidizing Bacteria in Syntrophy with Methanobacterium thermoautotrophicum or Methanobacterium thermoformicicum. AB - Thermophilic propionate-oxidizing, proton-reducing bacteria were enriched from the granular methanogenic sludge of a bench-scale upflow anaerobic sludge bed reactor operated at 55 degrees C with a mixture of volatile fatty acids as feed. Thermophilic hydrogenotrophic methanogens had a high decay rate. Therefore, stable, thermophilic propionate-oxidizing cultures could not be obtained by using the usual enrichment procedures. Stable and reproducible cultivation was possible by enrichment in hydrogen-pregrown cultures of Methanobacterium thermoautotrophicum DeltaH which were embedded in precipitates of FeS, achieved by addition of FeCl(2) to the media. The propionate-oxidizing bacteria formed spores which resisted pasteurization for 30 min at 90 degrees C or 10 min at 100 degrees C. Highly purified cultures were obtained with either M. thermoautotrophicum DeltaH or Methanobacterium thermoformicicum Z245 as the syntrophic partner organism. The optimum temperature for the two cultures was 55 degrees C. Maximum specific growth rates of cultures with M. thermoautotrophicum DeltaH were somewhat lower than those of cultures with M. thermoformicicum Z245 (0.15 and 0.19 day, respectively). Growth rates were even higher (0.32 day) when aceticlastic methanogens were present as well. M. thermoautotrophicum DeltaH is an obligately hydrogen-utilizing methanogen, showing that interspecies hydrogen transfer is the mechanism by which reducing equivalents are channelled from the acetogens to this methanogen. Boundaries of hydrogen partial pressures at which propionate oxidation occurred were between 6 and 34 Pa. Formate had a strong inhibitory effect on propionate oxidation in cultures with M. thermoautotrophicum. Inhibition by formate was neutralized by addition of the formate-utilizing methanogen or by addition of fumarate. Results indicate that formate inhibited succinate oxidation to fumarate, an intermediate step in the biochemical pathway of propionate oxidation. PMID- 16348633 TI - Isolation of 2,4-diacetylphloroglucinol from a fluorescent pseudomonad and investigation of physiological parameters influencing its production. AB - Pseudomonas sp. strain F113 was isolated from the rhizosphere of sugar beets and shown to inhibit a range of plant pathogenic fungi by producing an antibioticlike compound. An antibiotic-negative mutant strain, F113G22, was generated by transposon mutagenesis. This mutant has lost the ability to inhibit both bacterial and fungal microorganisms on high-iron medium. The antibioticlike compound was subsequently identified as 2,4-diacetylphloroglucinol (DAPG), and a high-pressure liquid chromatographic assay was developed for to detect it quantitatively in growth culture media and soil. The growth temperature had a direct bearing on DAPG production by strain F113, with maximum production at 12 degrees C. The iron concentration, pH, and oxygen had no influence on DAPG production by strain F113 under the assay conditions used. However, a low ratio of culture volume to surface area available to the microbe in the growth container was critical for optimum DAPG production. Different types of carbon sources influenced DAPG production by strain F113 to various degrees. For example, sucrose, fructose, and mannitol promoted high yields of DAPG by strain F113, whereas glucose and sorbose resulted in very poor DAPG production. PMID- 16348634 TI - Bacterial growth in the cold: evidence for an enhanced substrate requirement. AB - Growth responses and biovolume changes for four facultatively psychrophilic bacterial isolates from Conception Bay, Newfoundland, and the Arctic Ocean were examined at temperatures from - 1.5 to 35 degrees C, with substrate concentrations of 0.15, 1.5, and 1,500 mg of proteose peptone-yeast extract per liter. For two cultures, growth in 0.1, 1.0, and 1,000 mg of proline per liter was also examined. At 10 to 15 degrees C and above, growth rates showed no marked effect of substrate concentration, while at - 1.5 and 0 degrees C, there was an increasing requirement for organic nutrients, with generation times in low nutrient media that were two to three times longer than in high-nutrient media. Biovolume showed a clear dependence on substrate concentration and quality; the largest cells were in the highest-nutrient media. Biovolume was also affected by temperature; the largest cells were found at the lowest temperatures. These data have implications for both food web structure and carbon flow in cold waters and for the effects of global climate change, since the change in growth rate is most dramatic at the lowest temperatures. PMID- 16348635 TI - Influence of Calcium, Iron, and pH on Phosphate Availability for Microbial Mineralization of Organic Chemicals. AB - A study was conducted to determine some of the factors affecting the P requirement for the biodegradation of p-nitrophenol, phenol, and glucose by Pseudomonas and Corynebacterium strains. Mineralization of glucose was rapid and the Pseudomonas sp. grew extensively in solutions with 5 and 10 mM phosphate, but the rate and extent of degradation were low and the bacterial population never became abundant in media with 0.2 mM phosphate. Similar results were obtained with the Corynebacterium sp. growing in media containing p-nitrophenol or phenol and in solutions with a purified phosphate salt. The extent of growth of the Corynebacterium sp. was reduced with 2 or 10 mM phosphate in media containing high Fe concentrations. Ca at 5 mM but not 0.5 mM inhibited p-nitrophenol mineralization by the Corynebacterium sp. with phosphate concentrations from 0.2 to 5.0 mM. Phenol mineralization by the Pseudomonas sp. in medium with 0.2 mM phosphate was rapid at pH 5.2, but the bacteria had little or no activity at pH 8.0. In contrast, the activity was greater at pH 8.0 than at pH 5.2 when the culture contained 10 mM phosphate. These effects of pH were similar in media with 5 mM Ca or no added Ca. We conclude that the effect of P on bacterial degradation can be influenced by the pH and the concentrations of Fe and Ca. PMID- 16348636 TI - Anaerobic Production of Extracellular Polysaccharide by Butyrivibrio fibrisolvens nyx. AB - Anaerobic production of extracellular polysaccharide (EP) was examined, using a previously uncharacterized, obligately anaerobic rumen isolate, Butyrivibrio fibrisolvens nyx, which produced an EP that was rheologically similar to xanthan gum. The main objectives were to determine the nutritional requirements and conditions which promoted EP production by strain nyx. Strain nyx was grown anaerobically in defined and semidefined media. In addition to carbohydrate and nitrogen sources, strain nyx required acetic acid, folic acid, biotin, and pyridoxamine. Strain nyx produced similar amounts of EP at 35 to 40 degrees C. Conditions that improved growth usually improved EP production. Of the carbohydrates tested, glucose supported the fastest growth and most EP production, followed by sucrose, xylose, and lactose. Strain nyx utilized ammonium sulfate, urea, or vitamin-free casein hydrolysate as nitrogen sources for growth and EP production. At 2 and 20 g/liter, respectively, ammonium sulfate and vitamin-free casein hydrolysate supported about the same rates of growth and EP production. EP was not produced in the lag or stationary phases, and EP production was exponential during exponential cell growth. Based on the results of this work, anaerobic EP production with B. fibrisolvens nyx could reduce energy costs for industrial EP production compared with the cost of aerated systems. Finally, this work demonstrated that, under appropriate growth conditions, a gastrointestinal tract (ruminal) microorganism produced high levels of EP. PMID- 16348637 TI - Nitroaromatics Are Substrates for the TOL Plasmid Upper-Pathway Enzymes. AB - Expression of the xylMA genes encoding for toluene monoxygenase from the lactose promoter in a broad-host-range plasmid allows the oxidation of toluene and m- and p-nitrotoluene to their corresponding benzyl alcohols and benzaldehydes in Pseudomonas putida and Escherichia coli. Benzyl alcohols accumulate until reaching a concentration of about 80 muM, while benzaldehydes accumulate steadily with time for at least 24 h. TOL-encoded benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase recognize m- and p-nitro-substituted compounds as substrates. In contrast, the XylR protein, which regulates the TOL plasmid encoded upper-pathway operon, does not recognize nitro-substituted toluenes as effectors. PMID- 16348638 TI - Nitrate is a preferred electron acceptor for growth of freshwater selenate respiring bacteria. AB - An anaerobic, freshwater enrichment grew with either nitrate or selenate as an electron acceptor. With both ions present, nitrate reduction preceded selenate reduction. An isolate from the enrichment grew on either ion, but the presence of nitrate precluded the reduction of selenate. Stock cultures of denitrifiers grew anaerobically on nitrate but not on selenate. PMID- 16348639 TI - Transfer of IncP Plasmids to Extremely Acidophilic Thiobacillus thiooxidans. AB - The broad-host-range IncP plasmids RP4, R68.45, RP1::Tn501, and and pUB307 were transferred directly to extremely acidophilic Thiobacillus thiooxidans from Escherichia coli by conjugation at frequencies of 10 to 10 per recipient. The ability of T. thiooxidans to receive and express the antibiotic resistance markers was examined. The plasmid RP4 was transferred back to E. coli from T. thiooxidans at a frequency of 1.0 x 10 per recipient. PMID- 16348640 TI - Existence of a hydrogen sulfide:ferric ion oxidoreductase in iron-oxidizing bacteria. AB - The existence of a hydrogen sulfide:ferric ion oxidoreductase, which catalyzes the oxidation of elemental sulfur with ferric ions as an electron acceptor to produce ferrous and sulfite ions, was assayed with washed intact cells and cell extracts of various kinds of iron-oxidizing bacteria, such as Thiobacillus ferrooxidans 13598, 13661, 14119, 19859, 21834, 23270, and 33020 from the American Type Culture Collection, Leptospirillum ferrooxidans 2705 and 2391 from the Deutsche Sammlung von Mikroorganismen, L. ferrooxidans BKM-6-1339 and P3A, and moderately thermophilic iron-oxidizing bacterial strains BC1, TH3, and Alv. It was found that hydrogen sulfide:ferric ion oxidoreductase activity comparable to that of T. ferrooxidans AP19-3 was present in all iron-oxidizing bacteria tested, suggesting a wide distribution of this enzyme in iron-oxidizing bacteria. PMID- 16348641 TI - Diurnal Cycles of Sulfate Reduction under Oxic Conditions in Cyanobacterial Mats. AB - Diurnal cycles of sulfate reduction were examined in a well-developed cyanobacterial mat which grew in an outdoor experimental hypersaline pond system at a constant salinity of 75 +/- 5% for 3 years. Vertical profiles of sulfate reduction were determined for the upper 12 mm of the microbial mat. Sulfate reduction activities were compared with diurnal variations of oxygen and sulfide concentrations measured by microelectrodes. Significant activity of sulfate reducing bacteria was detected under aerobic conditions during the daytime, with maximal activity at 2 p.m. When comparing sulfate reduction activities in sediment cores taken at 6 a.m. and 12 a.m. and incubated at a constant temperature in the light and in the dark, a distinct stimulation of the activity in the vertical profile of sulfate reduction by light was evident. It is therefore concluded that the maximal in situ activities, measured at 2 p.m. in the chemocline of the cyanobacterial mat, cannot be attributed to diurnal changes of temperature alone. The response of sulfate-reducing bacteria to the addition of specific carbon sources was significantly different in the cyanobacterial layer, the anoxygenic phototrophic bacterial layer, and the permanently reduced layer of the microbial mat. Sulfate reduction in the mat layer exposed to high oxygen concentrations as a result of cyanobacterial oxygenic photosynthesis was enhanced only by glycolate; in the microzone where the chemocline is found during the daytime, ethanol was the only carbon source to enhance sulfate reduction, while both ethanol and lactate enhanced this activity in the permanently reduced zone. PMID- 16348642 TI - Evaluation of Leptospirillum ferrooxidans for Leaching. AB - The importance of Leptospirillum ferrooxidans for leach processes has been evaluated by studying the lithotrophic flora of three mine biotopes and a heap leaching operation, by percolation experiments with inoculated, sterilized ore, and by morphological, physiological, and genetic investigations of pure and mixed cultures of L. ferrooxidans, Thiobacillus ferrooxidans, and Thiobacillus thiooxidans. In biotopes of 20 degrees C or above, Leptospirillum-like bacteria are as abundant as T. ferrooxidans. Leptospirilli represent at least one-half of the ferrous-iron-oxidizing population. Percolation experiments confirmed this result. Leptospirilli were as numerous as T. ferrooxidans. At reduced temperatures, the generation times of leptospirilli increase more so than those of T. ferrooxidans. At 14 degrees C, Leptospirillum grows slowly and T. ferrooxidans dominates the population. Physiological investigations indicate that L. ferrooxidans is a strict chemolithoautotroph, metabolizing only ferrous iron and pyrite. Even an addition of 0.05% (wt/vol) yeast extract inhibited its growth. The maximum ferrous-iron-oxidizing activity of L. ferrooxidans amounts to about 40% of the activity of T. ferrooxidans. After growth on sulfidic ore, both species exhibit reduced iron-oxidizing activities, L. ferrooxidans exhibiting one third and T. ferrooxidans exhibiting one-seventh of their maximum activities. Surprisingly, the absolute values are similar. For indirect leaching, L. ferrooxidans is as important as T. ferrooxidans. This was confirmed by the results of percolation experiments. L. ferrooxidans together with T. thiooxidans mobilized metals at least as well as T. ferrooxidans did. The best results were obtained with a mixed culture of all three species. PMID- 16348643 TI - Stromal Development and Mating System of Balansia epichloe, a Leaf-Colonizing Endophyte of Warm-Season Grasses. AB - Studies of the stromal development and mating system of Balansia epichloe were conducted. Early development of the stroma consists of both endophytic and epiphytic phases of growth. As development progresses, the epiphytic stromal subiculum on the upper surface of leaves is linked with endophytic mycelium within leaves by hyphal bridges, which may provide carbohydrates for stromal development. Sugar utilization studies suggest that Balansia epichloe is excluded from growth within inflorescence primordia of grasses by the presence of high levels of sugars that are inhibitory to growth of the endophyte. Studies of the mating system of B. epichloe were conducted, and the fungus was shown to be heterothallic, with ephelidial conidia functioning as spermatia. Insect vectoring of ephelidial spermatia is suggested to account for the irregular pattern of perithecial development on stromata. PMID- 16348644 TI - Production of Concentrated Lactococcus lactis subsp. cremoris Suspensions in Calcium Alginate Beads. AB - The effect of simultaneous modification of medium composition and growth conditions on the production of Lactococcus lactis subsp. cremoris biomass in calcium alginate beads was studied by the response surface method. Statistical methods of data analysis for unbalanced experiments are illustrated. The media tested were whey, whey supplemented with yeast extract and/or meat extract, milk, and the commercial medium Gold Complete (Nordica). Fermentations were performed at 23 degrees C under pH control (5.6, 6.0, 6.4, or 6.8). In one complete series, 1% CaCO(3) was added to the growth media. There were strong interactions between CaCO(3) and media, CaCO(3) and pH level, and CaCO(3), media, and pH level. In media with CaCO(3), all first-order interactions between media, pH, and sampling time were significant. The addition of CaCO(3) increased cell counts in whey-meat extract medium, but no significant difference was found with the other media. Uncoupling between growth and acidification occurred between 16 and 22 h. Highest counts were obtained on milk and Gold Complete (6 x 10/g). In CaCO(3)-containing media, pH influenced cell counts only in whey and in Gold Complete (pH 5.6 and 6.0 giving the best results); pH also influenced the bead mass obtained at the end of the fermentation. Biomass production in alginate gels is proposed as a method of obtaining concentrated cell suspensions without centrifugation or filtration. PMID- 16348645 TI - Variation in microbial biomass and community structure in sediments of eutrophic bays as determined by phospholipid ester-linked Fatty acids. AB - The distribution of phospholipid ester-linked fatty acids (PLFA) in sediments of eutrophic bays (Hiroshima Bay and Aki Nada) was studied to quantify the microbial biomass, community structure, and nutritional status. A total of 63 fatty acids in the range of C(10) to C(24) were determined. They consist of saturated fatty acids, branched fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids, and variation was revealed in the relative proportions of these fatty acids in sediments. On the basis of the PLFA concentration in sediments, the calculated microbial biomass showed variation (mean +/- standard deviation = 0.70 x 10 +/- 0.53 x 10 cells per g [dry weight] of sediment) in the eutrophic bays. In sediments, a higher amount of biomass was observed in the coastal area of Hiroshima Bay than that observed in the rest of the bay and adjacent Aki Nada. The microbial community structure of the present study area, as characterized by the PLFA profiles, showed very low percentages of polyunsaturated fatty acids and long-chain fatty acids characteristic of microeukary-otes and terrestrial input, respectively, and high percentages of fatty acids characteristic of bacteria. The distribution of PLFA profiles also showed the relative contribution of both aerobic and anaerobic bacteria, especially sulfate-reducing bacteria, in the study area. The relative proportions of PLFA revealed distinctive differences among the stations of the study area, as is evidenced from six clusters obtained for the PLFA profiles. The results of Tukey's honestly significant difference test further confirmed that the sediments in the coastal area of Hiroshima Bay were significantly enriched by a number of fatty acids when compared with other areas investigated where relatively few fatty acids were present in significant quantities. No marked variation in environmental parameters in the surface- and bottom-water samples was observed, indicating the absence of any water movement in the study area. Furthermore, low redox potential and the levels of sulfide in the sediment revealed the reduced condition of the sediment. The existing environmental conditions and pollution of the study area were attributed to the observed microbial community structure in the sediments. PMID- 16348646 TI - Purification and Characterization of Extracellular Pectinolytic Enzymes Produced by Sclerotinia sclerotiorum. AB - An exopolygalacturonase (exoPG) and an exopolymethylgalacturonase (exoPMG) produced by Sclerotinia sclerotiorum have been purified by ammonium sulfate precipitation, gel filtration, and ion exchange chromatography. The exoPG and the exoPMG were purified 66- and 50-fold, respectively, by using a series of separation procedures that included ammonium sulfate precipitation and gel chromatography. Molecular masses of the native proteins were 68 kDa for exoPG and 140 kDa for exoPMG. The pH optima of the enzymes were about pH 5, and their optimum temperature was 45 degrees C. Activities of both enzymes were inhibited by Hg, Zn, Cu, and p-chloromercuribenzoate. ExoPMG activity, in contrast to exoPG activity, was stimulated by Mn and Co. ExoPMG hydrolyzed only citrus pectin, while exoPG degraded sodium polygalacturonate and, to a lesser extent, citrus pectin. The exo mode of action of the enzymes was revealed by thin-layer chromatography of substrate hydrolysates. Antibodies raised against each purified protein exhibited no cross-reaction, thus confirming the biochemical specificities of the enzymes. PMID- 16348647 TI - Effects of Temperature on Two Psychrophilic Ecotypes of a Heterotrophic Nanoflagellate, Paraphysomonas imperforata. AB - Two different psychrophilic types of the heterotrophic nanoflagellate Paraphysomonas imperforata were isolated from Newfoundland coastal waters and the Arctic Ocean. When fed bacteria without food limitation, both isolates were able to grow at temperatures from -1.8 to 20 degrees C, with maximum growth rates of 3.28 day at 15 degrees C and 2.28 day at 12.3 degrees C for the Newfoundland and the Arctic isolates, respectively. Ingestion rates increased with temperature from 14 to 62 bacteria flagellate h for the Newfoundland isolate and from 30 to 99 bacteria flagellate h for the Arctic isolate. While temperature did not affect cell yields (number of protozoa produced divided by number of bacteria consumed), it affected flagellate sizes. This differential effect of temperature on cell yield and cell size resulted in a changing gross growth efficiency (GGE) in terms of biovolume; colder temperatures favored higher GGEs. The comparison of Q(10) values for growth rates and ingestion rates between the isolates shows that the Arctic isolate is better adapted to extremely cold temperature than the Newfoundland isolate. At seawater-freezing temperature (-1.8 degrees C), the estimated maximum growth rates and maximum ingestion rates are 0.81 day and 30 bacteria flagellate h for the Arctic isolate and 0.54 day and 12 bacteria flagellate h for the Newfoundland isolate. Our findings about psychrophilic nanoflagellates fit the general characteristics of cold-water-dwelling organisms: reduced physiological rates and higher GGEs at lower temperatures. Because of the large and persistent differences between the isolates, we conclude that they are ecotypes adapted to specific environmental conditions. PMID- 16348648 TI - Bacterial oxidation of sulfide minerals in column leaching experiments at suboptimal temperatures. AB - The purpose of the work was to quantitatively characterize temperature effects on the bacterial leaching of sulfide ore material containing several sulfide minerals. The leaching was tested at eight different temperatures in the range of 4 to 37 degrees C. The experimental technique was based on column leaching of a coarsely ground (particle diameter, 0.59 to 5 mm) ore sample. The experimental data were used for kinetic analysis of chalcopyrite, sphalerite, and pyrrhotite oxidation. Chalcopyrite yielded the highest (73 kJ/mol) and pyrrhotite yielded the lowest (25 kJ/mol) activation energies. Especially with pyrrhotite, diffusion contributed to rate limitation. Arrhenius plots were also linear for the reciprocals of lag periods and for increases of redox potentials (dmV/dt). Mass balance analysis based on total S in leach residue was in agreement with the highest rate of leaching at 37 and 28 degrees C. The presence of elemental S in leach residues was attributed to pyrrhotite oxidation. PMID- 16348649 TI - Formation of Novel Polysaccharides by Bradyrhizobium japonicum Bacteroids in Soybean Nodules. AB - Certain strains of Bradyrhizobium japonicum form a previously unknown polysaccharide in the root nodules of soybean plants (Glycine max (L.) Merr.). The polysaccharide accumulates inside of the symbiosome membrane-the plant derived membrane enclosing the bacteroids. In older nodules (60 days after planting), the polysaccharide occupies most of the symbiosome volume and symbiosomes become enlarged so that there is little host cytoplasm in infected cells. The two different groups of B. japonicum which produce different types of polysaccharide in culture produce polysaccharides of similar composition in nodules. Polysaccharide formed by group I strains (e.g., USDA 5 and USDA 123) is composed of rhamnose, galactose, and 2-O-methylglucuronic acid, while polysaccharide formed by group II strains (e.g., USDA 31 and USDA 39) is composed of rhamnose and 4-O-methylglucuronic acid. That the polysaccharide is a bacterial product is indicated by its composition plus the fact that polysaccharide formation is independent of host genotype but is dependent on the bacterial genotype. Polysaccharide formation in nodules is common among strains in serogroups 123, 127, 129, and 31, with 27 of 39 strains (69%) testing positive. Polysaccharide formation in nodules is uncommon among other B. japonicum serogroups, with only 1 strain in 18 (6%) testing positive. PMID- 16348650 TI - Characterization of Fluorescent Siderophore-Mediated Iron Uptake in Pseudomonas sp. Strain M114: Evidence for the Existence of an Additional Ferric Siderophore Receptor. AB - In Pseudomonas sp. strain M114, the outer membrane receptor for ferric pseudobactin M114 was shown to transport ferric pseudobactins B10 and A225, in addition to its own. The gene encoding this receptor, which was previously cloned on pCUP3, was localized by Tn5 mutagenesis to a region comprising >1.6 kb of M114 DNA. A mutant (strain M114R1) lacking this receptor was then created by a marker exchange technique. Characterization of this mutant by using purified pseudobactin M114 in radiolabeled ferric iron uptake studies confirmed that it was completely unable to utilize this siderophore for acquisition of iron. In addition, it lacked an outer membrane protein band of 89 kDa when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. As a result, growth of the mutant was severely restricted under low-iron conditions. However, this phenotype was reversed in the presence of another fluorescent siderophore (pseudobactin MT3A) from Pseudomonas sp. strain MT3A, suggesting the presence of a second receptor in strain M114. Furthermore, wild-type Pseudomonas sp. strain B24 was not able to utilize ferric pseudobactin MT3A, and this phenotype was not reversed upon expression of the M114 receptor encoded on pCUP3. However, a cosmid clone (pMS1047) that enabled strain B24 to utilize ferric pseudobactin MT3A was isolated from an M114 gene bank. Radiolabel transport assays with purified pseudobactin MT3A confirmed this event. Plasmid pMS1047 was shown to encode an outer membrane protein of 81 kDa in strain B24 under iron-limiting conditions; this protein corresponds to a similar protein in strain M114. PMID- 16348651 TI - Persistence of viruses in desert soils amended with anaerobically digested sewage sludge. AB - Pima County, Ariz., is currently investigating the potential benefits of land application of sewage sludge. To assess risks associated with the presence of pathogenic enteric viruses present in the sludge, laboratory studies were conducted to measure the inactivation rate (k = log(10) reduction per day) of poliovirus type 1 and bacteriophages MS2 and PRD-1 in two sludge-amended desert agricultural soils (Brazito Sandy Loam and Pima Clay Loam). Under constant moisture (approximately -0.05 x 10 Pa for both soils) and temperatures of 15, 27, and 40 degrees C, the main factors controlling the inactivation of these viruses were soil temperature and texture. As the temperature increased from 15 to 40 degrees C, the inactivation rate increased significantly for poliovirus and MS2, whereas, for PRD-1, a significant increase in the inactivation rate was observed only at 40 degrees C. Clay loam soils afforded more protection to all three viruses than sandy soils. At 15 degrees C, the inactivation rate for MS2 ranged from 0.366 to 0.394 log(10) reduction per day in clay loam and sandy loam soils, respectively. At 27 degrees C, this rate increased to 0.629 log(10) reduction per day in clay loam soil and to 0.652 in sandy loam soil. A similar trend was observed for poliovirus at 15 degrees C (k = 0.064 log(10) reduction per day, clay loam; k = 0.095 log(10) reduction per day, sandy loam) and 27 degrees C (k = 0.133 log(10) reduction per day, clay loam; k = 0.154 log(10) reduction per day, sandy loam). Neither MS2 nor poliovirus was recovered after 24 h at 40 degrees C. No reduction of PRD-1 was observed after 28 days at 15 degrees C and after 16 days at 27 degrees C. At 40 degrees C, the inactivation rates were 0.208 log(10) reduction per day in amended clay loam soil and 0.282 log(10) reduction per day in sandy loam soil. Evaporation to less than 5% soil moisture completely inactivated all three viruses within 7 days at 15 degrees C, within 3 days at 27 degrees C, and within 2 days at 40 degrees C regardless of soil type. This suggests that a combination of high soil temperature and rapid loss of soil moisture will significantly reduce risks caused by viruses in sludge. PMID- 16348652 TI - Development and use of field application vectors to express nonadaptive foreign genes in competitive environments. AB - Many potential applications of genetically engineered microorganisms in environmental and agricultural biotechnology involve introducing genetic capabilities into nonsterile competitive environments in which they provide no advantage to the host. Field application vectors have been designed for the purpose of creating a temporary niche for the host in such environments. This technique involves the addition to the target environment of a selective substrate readily utilizable by the host microorganism but unavailable to most indigenous species. Thirteen nonionic and anionic detergents, representing a wide range of structural complexities and molecular weights, were screened as potential selective substrates. Competition experiments in soil, using Warburg respirometry, indicated that isolates from six different detergent enrichment cultures were more active on their corresponding detergents than the indigenous microorganisms. Detergents of intermediate structural complexities and molecular weights were most effective for use as selective substrates. A field application vector that utilizes 1.0% Igepal CO-720 (detergent) as the selective substrate and Pseudomonas paucimobilis 1IGP4 as the host was tested for its ability to increase the presence of nonadaptive tetracycline resistance marker genes in soil. In soil amended with the selective substrate, strain 1IGP4 plate counts increased by three orders of magnitude and tetracycline-resistant transformant (pRK293) counts increased from 1.8 x 10/g of soil to 4.3 x 10/g in 2 days. Inoculation in the absence of substrate amendment or amendment with a nonselective substrate did not result in growth of strain 1IGP4. These results demonstrate the effectiveness of field application vectors for increasing the concentration of nonadaptive genes in competitive environments. PMID- 16348653 TI - Total degradation of EDTA by mixed cultures and a bacterial isolate. AB - A bacterial mixed culture, which was obtained from sewage by a special enrichment procedure, utilized EDTA as the sole source of carbon and nitrogen for growth. High concentrations of mineral salts, particularly CaCl(2), or the use of a mineral base without nitrogen protected the cells from inactivation after transfer into fresh medium containing 200-mg/liter (0.67 mM) EDTA. The chemical speciation did not influence the biodegradability of EDTA. However, when resting cells of the mixed culture were incubated with EDTA in the presence of an equivalent molar amount of FeCl(3), the reaction came to a halt before the complete consumption of the substrate. A gram-negative isolate from the mixed population, BNC1, also metabolized EDTA in monoculture. Growth of the pure culture was promoted by biotin or folic acid but was always accompanied by the accumulation of unidentified metabolites and was slow (mu(max), 0.024 h) compared with that of the original community (mu(max), 0.036 h). PMID- 16348654 TI - Fluorimetric Detection of a Bacillus stearothermophilus Spore-Bound Enzyme, alpha d-Glucosidase, for Rapid Indication of Flash Sterilization Failure. AB - A biological indicator based on fluorimetric detection within 60 min of a Bacillus stearothermophilus spore-bound enzyme, alpha-d-glucosidase, has been developed. Results indicate that the enzyme survived slightly longer than spores observed after 24 h of incubation. The new system shows promise for evaluating flash sterilization cycles within 60 min compared with conventional 24-h systems. PMID- 16348655 TI - The Soybean Rj4 Allele Restricts Nodulation by Bradyrhizobium japonicum Serogroup 123 Strains. AB - Of nine Bradyrhizobium japonicum serogroup 123 strains examined, 44% were found to be restricted for nodulation by cultivar Hill. Nodulation studies with soybean isoline BARC-2 confirmed that the soybean Rj4 allele restricts nodulation by the same serogroup 123 isolates. Immunological analyses indicated that B. japonicum strains in serogroups 123 and 31 share at least one surface somatic antigen. PMID- 16348656 TI - Preparation of encapsulated microbial cells for environmental applications. AB - An improved method for the encapsulation of bacteria into microspheres of alginate, agarose, or polyurethane is described. Cell suspensions were passed through a low-pressure nozzle into an aqueous phase where matrix polymerization or gelation yielded beads 2 to 50 mum in diameter. Trials with a chlorophenol degrading Flavobacterium species showed that cells entrapped by these procedures were as catabolically active as free cells. These types of beads should have numerous applications in the fields of environmental science and engineering. PMID- 16348657 TI - Effect of Yeast Extract and Vitamin B(12) on Ethanol Production from Cellulose by Clostridium thermocellum I-1-B. AB - Addition to media of yeast extract, a vitamin mixture containing vitamin B(12), biotin, pyridoxamine, and p-aminobenzoic acid, or vitamin B(12) alone enhanced formation of ethanol but decreased lactate production in the fermentation of cellulose by Clostridium thermocellum I-1-B. A similar effect was not observed with C. thermocellum ATCC 27405 and JW20. PMID- 16348658 TI - Selective Medium for Quantitation of Bacillus popilliae in Soil and in Commercial Spore Powders. AB - A medium consisting of MYPGP agar supplemented with vancomycin was found to be highly selective for Bacillus popilliae, especially for strains originally isolated from Japanese beetle larvae. The medium has proven to be useful for the quantitation of B. popilliae spores in commercial spore powder and in soil. PMID- 16348659 TI - Use of inorganic membrane filters (anopore) for epifluorescence and scanning electron microscopy of nanoplankton and picoplankton. AB - Inorganic membrane filters (Anopore) were examined qualitatively by epifluorescence and scanning electron microscopy to determine their suitability for the study of nanoplankton and picoplankton. Compared with filters currently used, the Anopore filters allowed for increased resolution of the specimen with epifluorescence microscopy because of filter flatness and increased illumination caused by the large number of pores cm. The inorganic filters had a lower filtration rate than polycarbonate filters. For scanning electron microscopy, the metal oxide (Anopore) filters were efficient support for the plankton, with little charging of cells or background. PMID- 16348660 TI - Occurrence of Giardia and Cryptosporidium spp. in Surface Water Supplies. AB - [This corrects the article on p. 2610 in vol. 57.]. PMID- 16348661 TI - Comparative Study of 35 Bacteriophages of Lactobacillus helveticus: Morphology and Host Range. AB - This survey included 23 phages isolated from cheese whey and 12 temperate phages induced with mitomycin from their lysogenic host strains. All of the phages had an isometric head and a tail with a contractile sheath. In addition, short-tailed (160-nm-long) and long-tailed (260-nm-long) phages were distinguished. Short tailed phages were by far the most widespread in French cheese factories (32 of the 35 phages studied). The study of phage relationships enabled two large groups of strains to be distinguished: those not or slightly sensitive to phages and those very sensitive to phages. There was an obvious relationship in the first group between phage sensitivity (or resistance) and the geographic origin of the strains. The second group contained primarily strains from large international collections and those isolated from commercial starters. The relationships among short-tailed phages, either temperate or isolated as lytic, suggest that lysogenic strains could be the major source of phages in French cheese factories. PMID- 16348662 TI - Molecular Cloning of the Tryptophan Operon from an Aeromonas hydrophila Freshwater Isolate. AB - A genomic library of Aeromonas hydrophila F9 was constructed by using pBR322 as a vector. From that, two DNA fragments (5.8 and 11.6 kb) were isolated containing genetic information to complement trpA and trpB defects (5.8-kb fragment) and to complement trpA, trpB, trpC, trpD, and trpE defects (11.6-kb fragment) in Escherichia coli mutants. Evidence of the existence of a secondary promoter is given. PMID- 16348663 TI - Solid-phase products of bacterial oxidation of arsenical pyrite. AB - Bacterial leaching of an As-containing pyrite concentrate produced acidic (pH < 1) leachates. During the leaching, the bacteria solubilized both As and Fe, and these two elements were distributed in solution-phase and solid-phase products. Jarosite and scorodite were the exclusive crystalline products in precipitate samples from the bacterial leaching of the sulfide concentrate. PMID- 16348664 TI - Genetic Analysis of Rhizobium leguminosarum bv. Phaseoli Mutants Defective in Nodulation and Nodulation Suppression. AB - Nodulation-defective rhizobia and their nodule-forming derivatives containing cloned DNA from the wild type were used to study nodulation suppression in Phaseolus vulgaris L. Non-nitrogen-fixing derivatives which formed rhizobia containing white nodules induced partial suppression. Comparison of this with the complete suppression by Fix derivatives and a Fix mutant which formed rhizobia containing pink nodules suggests that the extent of suppression may be related to successive stages of nodule development. PMID- 16348665 TI - Hydrophobicities and electrophoretic mobilities of anaerobic bacterial isolates from methanogenic granular sludge. AB - The hydrophobicities and electrophoretic mobilities of isolates from methanogenic anaerobic granular sludge were measured and compared with those of strains from culture collections. All new isolates were highly hydrophobic, indicating that the upflow anaerobic sludge blanket reactor concept selects for hydrophobic bacteria. Methanothrix soehngenii, a methanogen often observed in methanogenic granular sludge, was highly hydrophobic and showed low electrophoretic mobility at pH 7. The role of this strain in the formation of methanogenic granular sludge is discussed. PMID- 16348666 TI - Detection of Renibacterium salmoninarum, the Causative Agent of Bacterial Kidney Disease in Salmonid Fish, from Pen-Cultured Coho Salmon. AB - The detection of Renibacterium salmoninarum antigen from pen-cultured coho salmon was attempted. Flounder (Limanda herzensteini) (n = 24), greenling (Hexagrammos otakii) (n = 5), Japanese sculpin (Cottus japonicus) (n = 1), and flathead (Platycephalus indicus) (n = 22) captured by fishing around coho salmon net pens were examined for the presence of R. salmoninarum antigen by an indirect dot blot assay and by an indirect fluorescent-antibody technique using polyclonal and monoclonal antibodies. R. salmoninarum antigen was detected from kidney samples of one greenling and six flathead. Moreover, 86 scallops (Patinopecten yessoensis) were hung from the edge of the net pen for 50 days, and R. salmoninarum antigen was demonstrated in 31 samples by the indirect dot blot assay and the indirect fluorescent-antibody technique. PMID- 16348667 TI - Natamycin as a fungicide in agar media. AB - Fungal inhibition in four commonly used agar media was improved by substituting natamycin (pimaricin) for cycloheximide. The recovery of bacteria was not affected by natamycin, whereas fungal contamination from a variety of samples was significantly suppressed. Furthermore, natamycin lacks the occupational health hazards of cycloheximide. Medium-dependent natamycin degradation occurred during the preparation and refrigerated storage of agar plates, but the addition of natamycin at 21.6 mug/ml resulted in effective residual activity. PMID- 16348668 TI - Isolation and Characterization of Haloanaerobacter chitinovorans gen. nov., sp. nov., a Halophilic, Anaerobic, Chitinolytic Bacterium from a Solar Saltern. AB - [This corrects the article on p. 260 in vol. 58.]. PMID- 16348669 TI - Influence of a supplemental carbon source on anaerobic dechlorination of pentachlorophenol in granular sludge. AB - [This corrects the article on p. 367 in vol. 58.]. PMID- 16348670 TI - Metabolic Engineering To Produce Tyrosine or Phenylalanine in a Tryptophan Producing Corynebacterium glutamicum Strain. AB - The aromatic amino acids are synthesized via a common biosynthetic pathway. A tryptophan-producing mutant of Corynebacterium glutamicum was genetically engineered to produce tyrosine or phenylalanine in abundance. To achieve this, three biosynthetic genes encoding the first enzyme in the common pathway, 3-deoxy d-arabino-heptulosonate 7-phosphate synthase (DS), and the branch-point enzymes chorismate mutase and prephenate dehydratase were individually cloned from regulatory mutants of C. glutamicum which have either of the corresponding enzymes desensitized to end product inhibition. These cloned genes were assembled one after another onto a multicopy vector of C. glutamicum to yield two recombinant plasmids. One plasmid, designated pKY1, contains the DS and chorismate mutase genes, and the other, designated pKF1, contains all three biosynthetic genes. The enzymes specified by both plasmids were simultaneously overexpressed approximately sevenfold relative to the chromosomally encoded enzymes in a C. glutamicum strain. When transformed with pKY1 or pKF1, tryptophan producing C. glutamicum KY10865, with the ability to produce 18 g of tryptophan per liter, was altered to produce a large amount of tyrosine (26 g/liter) or phenylalanine (28 g/liter), respectively, because the accelerated carbon flow through the common pathway was redirected to tyrosine or phenylalanine. PMID- 16348671 TI - Three Genes for Metabolism of the Phytoalexin Maackiain in the Plant Pathogen Nectria haematococca: Meiotic Instability and Relationship to a New Gene for Pisatin Demethylase. AB - Some isolates of the plant-pathogenic fungus Nectria haematococca mating population (MP) VI metabolize maackiain and medicarpin, two antimicrobial compounds (phytoalexins) synthesized by chickpea (Cicer arietinum L.). The enzymatic modifications by the fungus convert the phytoalexins to less toxic derivatives, and this detoxification has been proposed to be important for pathogenesis on chickpea. In the present study, loci controlling maackiain metabolism (Mak genes) were identified by crosses among isolates of N. haematococca MP VI that differed in their ability to metabolize the phytoalexin. Strains carrying Mak1 or Mak2 converted maackiain to 1a-hydroxymaackiain, while those with Mak3 converted it to 6a-hydroxymaackiain. Mak1 and Mak2 were unusual in that they often failed to be inherited by progeny. Mak1 was closely linked to Pda6, a new member in a family of genes in N. haematococca MP VI that encode enzymes for detoxification of pisatin, the phytoalexin synthesized by garden pea. Like Mak1, Pda6 was also transmitted irregularly to progeny. Although the unusual meiotic behaviors of some Mak genes complicate genetic analysis, identification of these genes should afford a more through evaluation of the role of phytoalexin detoxification in the pathogenesis of N. haematococca MP VI on chickpea. PMID- 16348672 TI - Genetic Analysis of the Role of Phytoalexin Detoxification in Virulence of the Fungus Nectria haematococca on Chickpea (Cicer arietinum). AB - Chickpea (Cicer arietium L.) produces the antimicrobial compounds (phytoalexins) medicarpin and maackiain in response to infection by microorganisms. Nectria haematococca mating population (MP) VI, a fungus pathogenic on chickpea, can metabolize maackiain and medicarpin to less toxic products. These reactions are thought to be detoxification mechanisms in N. haematococca MP VI and required for pathogenesis by this fungus on chickpea. In the present study, these hypotheses were tested by examining the phenotypes of progeny from crosses of the fungus that segregated for genes (Mak genes) controlling phytoalexin metabolism. Mak1 and Mak2, two genes that individually confer the ability to convert maackiain to its 1a-hydroxydienone derivative, were linked to higher tolerance of the phytoalexins and high virulence on chickpea. These results indicate that this metabolic reaction is a mechanism for increased phytoalexin tolerance in the fungus, which thereby allows a higher virulence on chickpea. Mak3, a gene conferring the ability to convert maackiain to its 6a-hydroxypterocarpan derivative, also increased tolerance to maackiain in strains which carried it; however, the contribution of Mak3 to the overall level of pathogenesis could not be evaluated because most progeny from the cross segregating for this gene were low in virulence. Thus, metabolic detoxification of phytoalexins appeared to be necessary, as demonstrated in the Mak1 and Mak2 crosses, but not sufficient by itself, as in the Mak3 cross, for high virulence of N. haematococca MP VI on chickpea. PMID- 16348673 TI - Extensive In Vitro Hyphal Growth of Vesicular-Arbuscular Mycorrhizal Fungi in the Presence of CO(2) and Flavonols. AB - Various flavonoids were tested for their ability to stimulate in vitro growth of germinated spores of vesicular-arbuscular mycorrhizal fungi. Experiments were performed in the presence of 2% CO(2), previously demonstrated to be required for growth of Gigaspora margarita (G. Becard and Y. Piche, Appl. Environ. Microbiol. 55:2320-2325, 1989). Only the flavonols stimulated fungal growth. The flavones, flavanones, and isoflavones tested were generally inhibitory. Quercetin (10 muM) prolonged hyphal growth from germinated spores of G. margarita from 10 to 42 days. An average of more than 500 mm of hyphal growth and 13 auxiliary cells per spore were obtained. Quercetin also stimulated the growth of Glomus etunicatum. The glycosides of quercetin, rutin, and quercitrin were not stimulatory. The axenic growth of G. margarita achieved here under rigorously defined conditions is the most ever reported for a vesicular-arbuscular mycorrhizal fungus. PMID- 16348674 TI - Construction of Novel Bacillus thuringiensis Strains with Different Insecticidal Activities by Transduction and Transformation. AB - The shuttle vector pHT3101 and its derivative pHT408, bearing a copy of a cryIA(a) delta-endotoxin gene, were transferred into several Bacillus thuringiensis subspecies through phage CP-54Ber-mediated transduction, with frequencies ranging from 5 x 10 to 2 x 10 transductant per CFU, depending on the strain and on the plasmid. In Cry and Cry native recipients, the introduction of the cryIA(a) gene resulted in the formation of large bipyramidal crystals that were active against the insect Plutella xylostella (order Lepidoptera). In both cases, high levels of gene expression were observed. Transductants displaying a dual specificity were constructed by using as recipients the new isolates LM63 and LM79, which have larvicidal activity against insects of the order Coleoptera. It was not possible, however, to introduce pHT7911 into B. thuringiensis subsp. entomocidus, aizawai, or israelensis by transduction. However, electrotransformation was successful, and transformants expressing the toxin gene cryIIIA, carried by pHT7911, were obtained. Again, high levels of expression of the cloned gene were observed. The results indicate that CP-54Ber-mediated transduction is a useful procedure for introducing cloned crystal protein genes into various B. thuringiensis recipients and thereby creating strains with new combinations of genes. Finally it was also shown that pHT3101 is a very good expression vector for the cloned delta-endotoxin genes in the different recipients. PMID- 16348675 TI - Accumulation of Ergopeptide Alkaloids in Symbiotic Tall Fescue Grown under Deficits of Soil Water and Nitrogen Fertilizer. AB - The fungus Acremonium coenophialum is endophytically associated with tall fescue (Festuca arundinacea Schreber). Within this symbiotum the fungus produces ergopeptide alkaloids, which are associated with livestock toxicoses. Environmental effects on the production of ergot alkaloids within the symbiotum are unknown. We conducted a greenhouse study of the effects of flooding, nitrogen rate during fertilization (11, 73, and 220 mg of N per pot weekly), nitrogen form (3.4 and 34 mg of N as NH(4) or NO(3) per pot), and drought stress (-0.03, -0.05, and -0.50 MPa) on ergopeptide alkaloid concentrations in one genotype of nonsymbiotic and symbiotic tall fescue grown in plastic pots. It was determined that the concentration of ergovaline, the major type of ergopeptide alkaloid, was increased but was not as high as that in nonflooded controls. Total ergopeptide and ergovaline concentrations in plants receiving high (220 mg of N per pot) and low (11 mg of N per pot) levels of NH(4)NO(3) fertilization were not affected by flooding. The form of nitrogen was important since all concentrations of NO(3)-N increased ergopeptide alkaloid content, as opposed to the effects of NH(4)-N, which was effective only at high concentrations (34 mg of N per pot). Ergopeptide concentrations were highest in drought-stressed plants grown at -0.50 MPa and fertilized at the moderate or high N rate. The results suggest that within this genotype, ergopeptide alkaloid biosynthesis by the fungus is not appreciably affected by flooding but is greatly increased by high rates of N fertilization and moderate water deficit. PMID- 16348676 TI - Isolation of new aureobasidium strains that produce high-molecular-weight pullulan with reduced pigmentation. AB - New isolates of Aureobasidium pullulans were obtained from plant leaf surfaces gathered in San Diego County. The new fungal isolates were identified as A. pullulans on the basis of the appearance of polymorphic colonies formed on agar plates, the electrophoretic profiles of repeated genomic DNA sequences, and the production of pullulan in shake flask cultures. The isolates showed different degrees of pigmentation. One of the natural isolates was nonpigmented under mock production conditions in liquid culture, but was still able to synthesize a reduced amount of pigment on agar plates at late times. A mutagenic treatment with ethidium bromide produced derivatives of normally pigmented natural isolates that exhibited an increased tendency toward yeastlike growth and reduced pigmentation. Additionally, some of the new isolates and mutant derivatives accumulated pullulan of relatively high molecular weight in the culture broths. PMID- 16348677 TI - Conversion of Pyruvate to Acetoin Helps To Maintain pH Homeostasis in Lactobacillus plantarum. AB - Pyruvate is the substrate for diacetyl and acetoin synthesis by lactobacilli. Exogenous pyruvate stimulates acetoin production when glucose is present as an energy source. In Lactobacillus plantarum ATCC 8014, the energy derived from glucose via glycolysis generated a constant proton motive force of about -120 mV. At a low external pH, energized cells rapidly transported and accumulated pyruvate but did not do so when they were deenergized by nigericin. When large amounts of pyruvate were transported and subsequently accumulated internally, the cotransported protons rapidly lowered the internal pH. The conversion of pyruvate to acetoin instead of acidic end products contributed to the maintenance of pH homeostasis. This is the first report showing that the conversion of pyruvate to acetoin serves as a mechanism of pH homeostasis. PMID- 16348678 TI - Detection of the Light Organ Symbiont, Vibrio fischeri, in Hawaiian Seawater by Using lux Gene Probes. AB - Symbiotic bacteria that inhabit the light-emitting organ of the Hawaiian squid Euprymna scolopes are distinctive from typical Vibrio fischeri organisms in that they are not visibly luminous when grown in laboratory culture. Therefore, the abundance of these bacteria in seawater samples cannot be estimated simply by identifying them among luminous colonies that arise on nutrient agar plates. Instead, we have used luxR and polymerase chain reaction generated luxA gene probes to identify both luminous and non-visibly luminous V. fischeri colonies by DNA-DNA hybridization. The probes were specific, hybridizing at least 50 to 100 times more strongly to immobilized DNAs from V. fischeri strains than to those of pure cultures of other related species. Thus, even non-visibly luminous V. fischeri colonies could be identified among colonies obtained from natural seawater samples by their probe-positive reaction. Bacteria in seawater samples, obtained either within or distant from squid habitats, were collected on membrane filters and incubated until colonies appeared. The filters were then observed for visibly luminous V. fischeri colonies and hybridized with the lux gene probes to determine the number of total V. fischeri colonies (both luminous and non-visibly luminous). We detected no significant differences in the abundance of luminous V. fischeri CFU in any of the water samples observed (425 nm to damage fecal coliforms was dependent on the presence of dissolved sensitizers. The sensitizers were ubiquitous in raw sewage, unaffected by sewage treatment, not derivatives of bacteriochlorophyll or chlorophyll, absorbed well in UV light, and had a slight yellowish color; they are therefore believed to be humic substances. The ability of light to damage fecal coliforms was highly sensitive to, and completely dependent on, oxygen. Scavengers of H(2)O(2) and singlet oxygen could protect the bacteria from the effects of sunlight, but scavengers of hydroxyl radicals and superoxides could not. Light mediated damage of fecal coliforms was highly sensitive to elevated pH values, which also enabled light with wavelengths of >425 nm (in the presence of the sensitizer) to damage the bacteria. We conclude that humic substances, pH, and dissolved oxygen are important variables in the process by which light damages microorganisms in this and other environments and that these variables should be considered in future research on, and models of, the effects of light. PMID- 16348699 TI - Distribution, Frequency, and Diversity of Bacillus thuringiensis in an Animal Feed Mill. AB - Bacillus thuringiensis was isolated from 36 of 50 residue samples obtained from an animal feed mill (a stored-product environment). Of 710 selected colonies having Bacillus cereus-B. thuringiensis morphology isolated from the samples, 477 were classified as B. thuringiensis because of production of parasporal delta endotoxin crystals. There was a diverse population of B. thuringiensis, as revealed by differentiation of the isolates into 36 subgroups by using (i) their spectra of toxicity to the lepidopterans Heliothis virescens, Pieris brassicae, and Spodoptera littoralis and the dipteran Aedes aegypti and (ii) their parasporal crystal morphology. A total of 55% of the isolates were not toxic to any of these insects at the concentrations used in the bioassays; 40% of all isolates were toxic to one or more of the Lepidoptera; and 20, 1, and 1% of the isolates were toxic to only P. brassicae, H. virescens, and S. littoralis, respectively. The most frequent toxicity was toxicity to P. brassicae (36% of all isolates); 18% of the isolates were toxic to A. aegypti (5% exclusively), 10% were toxic to H. virescens, and 4% were toxic to S. littoralis. Toxicity to P. brassicae was more often linked with toxicity to H. virescens than with toxicity to S. littoralis. The frequency of toxicity was significantly greater in isolates that produced bipyramidal crystals than in isolates that produced irregular pointed, irregular spherical, rectangular, or spherical crystals. PMID- 16348700 TI - Metabolism of phosphonoacetate as the sole carbon and phosphorus source by an environmental bacterial isolate. AB - A gram-negative bacterium isolated from activated sludge was able to utilize up to 25 mM phosphonoacetate as the sole carbon and phosphorus source, with simultaneous excretion of virtually equimolar levels of phosphate. 2 Aminoethylphosphonate was similarly utilized with equivalent growth rates and cellular yields, while 3-aminopropyl-, 4-aminobutyl-, methyl-, ethyl-, and phenylphosphonates served only as phosphorus sources. PMID- 16348701 TI - Bacterial hydrolysis of protein and methylated protein and its implications for studies of protein degradation in aquatic systems. AB - Ribulose 1,5-bisphosphate carboxylase was radiolabelled by in vitro translation, resulting in uniformly labelled ribulose 1,5-bisphosphate carboxylase, and also by reductive methylation. We investigated the degradation of the two forms of radiolabelled protein by natural bacterial populations. Although total hydrolysis of uniformly labelled protein and methylated protein was nearly equal, percent assimilation, respiration, and release as low-molecular-weight material were different. Radioactivity from uniformly labelled protein was approximately equally assimilated into cells, respired as H(2)O, and released as low-molecular weight material, but radioactivity from the methylated protein was nearly all released as low-molecular-weight material, and little was assimilated or respired. PMID- 16348702 TI - Enzymic Dehalogenation of 4-Chlorobenzoyl Coenzyme A in Acinetobacter sp. Strain 4-CB1. AB - 4-Chlorobenzoate degradation in cell extracts of Acinetobacter sp. strain 4-CB1 occurs by initial synthesis of 4-chlorobenzoyl coenzyme A (4-chlorobenzoyl CoA) from 4-chlorobenzoate, CoA, and ATP. 4-Chlorobenzoyl CoA is dehalogenated to 4 hydroxybenzoyl CoA. Following the dehalogenation reaction, 4-hydroxybenzoyl CoA is hydrolyzed to 4-hydroxybenzoate and CoA. Possible roles for the CoA moiety in the dehalogenation reaction are discussed. PMID- 16348703 TI - Stimulation of reductive dechlorination of tetrachloroethene in anaerobic aquifer microcosms by addition of short-chain organic acids or alcohols. AB - The effect of the addition of common fermentation products on the dehalogenation of tetrachloroethene was studied in methanogenic slurries made with aquifer solids. Lactate, propionate, crotonate, butyrate, and ethanol stimulated dehalogenation activity, while acetate, methanol, and isopropanol did not. PMID- 16348704 TI - Factors stimulating propagation of legionellae in cooling tower water. AB - Our survey of cooling tower water demonstrated that the highest density of legionellae, >/=10 CFU/100 ml, appeared in water containing protozoa, >/=10 MPN/100 ml, and heterotrophic bacteria, >/=10 CFU/100 ml, at water temperatures between 25 and 35 degrees C. Viable counts of legionellae were detected even in the winter samples, and propagation, up to 10 CFU/100 ml, occurs in summer. The counts of legionellae correlated positively with increases in water temperature, pH, and protozoan counts, but not with heterotrophic bacterial counts. The water temperature of cooling towers may promote increases in the viable counts of legionellae, and certain microbes, e.g., protozoa or some heterotrophic bacteria, may be a factor stimulating the propagation of legionellae. PMID- 16348705 TI - Methanogenic degradation of poly(3-hydroxyalkanoates). AB - Poly(3-hydroxybutyrate) and the copolymer poly(3-hydroxybutyrate-co-3 hydroxyvalerate) were fermented to methane and carbon dioxide within 16 days by an anaerobic sewage sludge consortium. The cultures adapted quickly to metabolize these polymeric compounds, and between 83 and 96% of the substrate carbon was transformed to methane and carbon dioxide. PMID- 16348706 TI - Bdellovibrios in Callinectus sapidus, the Blue Crab. AB - Bdellovibrios were recovered from the gill tissue of all of 31 crabs sampled and from all samples of epibiota obtained from the ventral shell surface of 15 crabs. The results suggest that the blue crab is a reservoir for bdellovibrios. The association with crabs may be an important factor in the ecology of the bdellovibrios. PMID- 16348707 TI - Variation in microbial biomass and community structure in sediments of eutrophic bays as determined by phospholipid ester-linked Fatty acids. AB - [This corrects the article on p. 562 in vol. 58.]. PMID- 16348708 TI - Reciprocal isomerization of butyrate and isobutyrate by the strictly anaerobic bacterium strain WoG13 and methanogenic isobutyrate degradation by a defined triculture. AB - Isomerization of butyrate and isobutyrate was investigated with the recently isolated strictly anaerobic bacterium strain WoG13 which ferments glutarate to butyrate, isobutyrate, CO(2), and small amounts of acetate. Dense cell suspensions converted butyrate to isobutyrate and isobutyrate to butyrate. C nuclear magnetic resonance experiments proved that this isomerization was accomplished by migration of the carboxyl group to the adjacent carbon atom. In cell extracts, both butyrate and isobutyrate were activated to their coenzyme A (CoA) esters by acyl-CoA:acetate CoA-transferases. The reciprocal rearrangement of butyryl-CoA and isobutyryl-CoA was catalyzed by a butyryl-CoA:isobutyryl-CoA mutase which depended strictly on the presence of coenzyme B(12). Isobutyrate was completely degraded via butyrate to acetate and methane by a defined triculture of strain WoG13, Syntrophomonas wolfei, and Methanospirillum hungatei. PMID- 16348709 TI - Effect of growth temperature on fatty acid composition of ten thermus strains. AB - The fatty acid composition of Thermus spp., including T. aquaticus ATCC 25104, T. thermophilus DSM 579, T. flavus DSM 674, and seven wild strains was examined. Organisms were tested at a minimum of either 35, 40, or 45 degrees C and at an optimum of 60 or 70 degrees C. Total fatty acid content per dry weight of cells varied between 1.2 and 3.7%, and the quantity of fatty acids was higher at the high temperature range in the majority of strains. At the optimum temperature, strains could be assigned to three chemotaxonomic groups with reference to the ratio of iso C(15:0)/iso C(17:0). In six of the strains the ratio of iso C(15:0)/iso C(17:0) remained unchanged at the minimum temperature, whereas in four strains the ratio was reversed. The proportion of the C(15:0) and C(17:0) isobranched acids was decreased and the proportion of anteisobranched fatty acids, namely anteiso C(15:0), anteiso C(17:0), and anteiso C(17:1), was increased at the lower temperature range. Some changes were seen in the levels of the n-C(16:0) and iso C(16:0) acids, but these were strain specific. PMID- 16348710 TI - Molybdenum oxidation by Thiobacillus ferrooxidans. AB - Thiobacillus ferrooxidans AP19-3 oxidized molybdenum blue (Mo) enzymatically. Molybdenum oxidase in the plasma membrane of this bacterium was purified ca. 77 fold compared with molybdenum oxidase in cell extract. A purified molybdenum oxidase showed characteristic absorption maxima due to reduced-type cytochrome oxidase at 438 and 595 nm but did not show absorption peaks specific for c-type cytochrome. The optimum pH of molybdenum oxidase was 5.5. The activity of molybdenum oxidase was completely inhibited by sodium cyanide (5 mM) or carbon monoxide, and an oxidized type of cytochrome oxidase in a purified molybdenum oxidase was reduced by molybdenum blue, indicating that cytochrome oxidase in the enzyme plays a crucial role in molybdenum blue oxidation. PMID- 16348716 TI - Preparation of Refractile Spores of Clostridium thermosaccharolyticum Involves a Solventogenic Phase. AB - Conversion of vegetative cells of Clostridium thermosaccharolyticum to refractile endospores was achieved by sequential transfer and dilution at each generation, with a final dilution into a sporulation medium that contained xylan supplemented with excess calcium. The subsequent growth was synchronous and resulted in elongated, solventogenic cells that were then shifted to 35 degrees C to permit further differentiation without cell division. The synchronized cells grown in xylan medium supplemented with Ca gluconate produced total solvents that reached 9.63% (vol/vol). One hundred percent of these elongated solventogenic cells (4.84 x 10 cells per ml) entered the sporangial stage and continued to differentiate into refractile spores. Only cells sequentially transferred and diluted at a critical time of the growth cycle are synchronized, induced to elongate (>/=fourfold), become highly solventogenic in the presence of excess calcium, and are converted to a homogeneous population of refractile spores. PMID- 16348717 TI - Specific and quantitative assessment of naphthalene and salicylate bioavailability by using a bioluminescent catabolic reporter bacterium. AB - A bioassay was developed and standardized for the rapid, specific, and quantitative assessment of naphthalene and salicylate bioavailability by use of bioluminescence monitoring of catabolic gene expression. The bioluminescent reporter strain Pseudomonas fluorescens HK44, which carries a transcriptional nahG-luxCDABE fusion for naphthalene and salicylate catabolism, was used. The physiological state of the reporter cultures as well as the intrinsic regulatory properties of the naphthalene degradation operon must be taken into account to obtain a high specificity at low target substrate concentrations. Experiments have shown that the use of exponentially growing reporter cultures has advantages over the use of carbon-starved, resting cultures. In aqueous solutions for both substrates, naphthalene and salicylate, linear relationships between initial substrate concentration and bioluminescence response were found over concentration ranges of 1 to 2 orders of magnitude. Naphthalene could be detected at a concentration of 45 ppb. Studies conducted under defined conditions with extracts and slurries of experimentally contaminated sterile soils and identical uncontaminated soil controls demonstrated that this method can be used for specific and quantitative estimations of target pollutant presence and bioavailability in soil extracts and for specific and qualitative estimations of napthalene in soil slurries. PMID- 16348718 TI - Leaching of Pyrites of Various Reactivities by Thiobacillus ferrooxidans. AB - Wide variations were found in the rate of chemical and microbiological leaching of iron from pyritic materials from various sources. Thiobacillus ferrooxidans accelerated leaching of iron from all of the pyritic materials tested in shake flask suspensions at loadings of 0.4% (wt/vol) pulp density. The most chemically reactive pyrites exhibited the fastest bioleaching rates. However, at 2.0% pulp density, a delay in onset of bioleaching occurred with two of the pyrites derived from coal sources. T. ferrooxidans was unable to oxidize the most chemically reactive pyrite at 2.0% pulp density. No inhibition of pyrite oxidation by T. ferrooxidans occurred with mineral pyrite at 2.0% pulp density. Experiments with the most chemically reactive pyrite indicated that the leachates from the material were not inhibitory to iron oxidation by T. ferrooxidans. PMID- 16348719 TI - Sequential Transhydroxylations Converting Hydroxyhydroquinone to Phloroglucinol in the Strictly Anaerobic, Fermentative Bacterium Pelobacter massiliensis. AB - The recently isolated fermenting bacterium Pelobacter massiliensis is the only strict anaerobe known to grow on hydroxyhydroquinone (1,2,4-trihydroxybenzene) as the sole source of carbon and energy, converting it to stoichiometric amounts of acetate. In this paper, we report on the enzymatic reactions involved in the conversion of hydroxyhydroquinone and pyrogallol (1,2,3-trihydroxybenzene) to phloroglucinol (1,3,5-trihydroxybenzene). Cell extracts of P. massiliensis transhydroxylate pyrogallol to phloroglucinol after addition of 1,2,3,5 tetrahydroxybenzene (1,2,3,5-TTHB) as cosubstrate in a reaction identical to that found earlier with Pelobacter acidigallici (A. Brune and B. Schink, J. Bacteriol. 172:1070-1076, 1990). Hydroxyhydroquinone conversion to phloroglucinol is initiated in cell extracts without an external addition of cosubstrates. It involves a minimum of three consecutive transhydroxylation reactions characterized by the transient accumulation of two different TTHB isomers. Chemical synthesis of the TTHB intermediates allowed the resolution of the distinct transhydroxylation steps in this sequence. In an initial transhydroxylation, the hydroxyl group in the 1-position of a molecule of hydroxyhydroquinone is transferred to the 5-position of another molecule of hydroxyhydroquinone to give 1,2,4,5-TTHB and resorcinol (1,3-dihydroxybenzene) as products. Following this disproportionation of hydroxyhydroquinone, the 1,2,4,5 isomer is converted to 1,2,3,5-TTHB, an enzymatic activity present only in hydroxyhydroquinone-grown cells. Finally, phloroglucinol is formed from 1,2,3,5 TTHB by transfer of the 2-hydroxyl group to either hydroxyhydroquinone or resorcinol. The resulting coproducts are again cosubstrates in earlier reactions of this sequence. From the spectrum of hydroxybenzenes transhydroxylated by the cell extracts, the minimum structural prerequisites that render a hydroxybenzene a hydroxyl donor or acceptor are deduced. PMID- 16348720 TI - Genetic and Phenotypic Diversity of Bacillus polymyxa in Soil and in the Wheat Rhizosphere. AB - Diversity among 130 strains of Bacillus polymyxa was studied; the bacteria were isolated by immunotrapping from nonrhizosphere soil (32 strains), rhizosphere soil (38 strains), and the rhizoplane (60 strains) of wheat plantlets growing in a growth chamber. The strains were characterized phenotypically by 63 auxanographic (API 50 CHB and API 20B strips) and morphological features, serologically by an enzyme-linked immunosorbent assay, and genetically by restriction fragment length polymorphism (RFLP) profiles of total DNA in combination with hybridization patterns obtained with an rRNA gene probe. Cluster analysis of phenotypic characters by the unweighted pair group method with averages indicated four groups at a similarity level of 93%. Clustering of B. polymyxa strains from the various fractions showed that the strains isolated from nonrhizosphere soil fell into two groups (I and II), while the third group (III) mainly comprised strains isolated from rhizosphere soil. The last group (IV) included strains isolated exclusively from the rhizoplane. Strains belonging to a particular group exhibited a similarity level of 96%. Serological properties revealed a higher variability among strains isolated from nonrhizosphere and rhizosphere soil than among rhizoplane strains. RFLP patterns also revealed a greater genetic diversity among strains isolated from nonrhizosphere and rhizosphere soil and therefore could not be clearly grouped. The RFLP patterns of sorbitol-positive strains isolated from the rhizoplane were identical. These results indicate that diversity within populations of B. polymyxa isolated from nonrhizosphere and rhizosphere soil is higher than that of B. polymyxa isolated from the rhizoplane. It therefore appears that wheat roots may select a specific subpopulation from the soil B. polymyxa population. PMID- 16348721 TI - Enumeration of Thiobacilli within pH-Neutral and Acidic Mine Tailings and Their Role in the Development of Secondary Mineral Soil. AB - The Lemoine tailings of Chibougamau, Quebec, Canada, were deposited as a pH neutral mineral conglomerate consisting of aluminum-silicates, iron-aluminum silicates, pyrite, chalcopyrite, and sphalerite. These tailings are colonized by an active population of Thiobacillus ferrooxidans which is localized to an acid zone occupying 40% of the tailings' surface. This population peaked at 7 x 10 most probable number per gram of tailings during July and August 1990 and extended to a depth of 40 cm from the surface. Examination of samples over this depth profile by transmission electron microscopy and electron dispersive spectroscopy revealed a microbially mediated mineral transition from sulfides (below 40 cm) to chlorides and phosphates (at the surface). Silicate minerals were unaltered by microbial action. Transmission electron microscopy showed a tight association between Thiobacillus species and the sulfide minerals, which helps account for their prominence in tailings environments. Accurate enumeration of T. ferrooxidans from tailings required the disruption of their bonding to the mineral interface. Vortexing of a 10% aqueous suspension of the tailings material prior to most-probable-number analysis best facilitated this release. Even though heavy metals were highly mobile under acidic conditions at the Lemoine tailings, it was evident by transmission electron microscopy and electron dispersive spectroscopy that they were being immobilized as bona fide fine-grain minerals containing iron, copper, chlorine, phosphorus, and oxygen on bacterial surfaces and exopolymers. This biomineralization increased with increasing bacterial numbers and was most evident in the upper 3 cm of the acidic zone. PMID- 16348722 TI - Effects of Grazing by Flagellates on Competition for Ammonium between Nitrifying and Heterotrophic Bacteria in Chemostats. AB - The enhanced mineralization of organic nitrogen by bacteriophagous protozoa is thought to favor the nitrification process in soils, in which nitrifying bacteria have to compete with heterotrophic bacteria for the available ammonium. To obtain more insight into this process, the influence of grazing by the bacteriovorous flagellate Adriamonas peritocrescens on the competition for limiting amounts of ammonium between the ammonium-oxidizing species Nitrosomonas europaea and the heterotrophic species Arthrobacter globiformis was studied in the presence of Nitrobacter winogradskyi in continuous cultures at dilution rates of 0.004 and 0.01 h. The ammonium concentration in the reservoir was maintained at 2 mM, whereas the glucose concentration was increased stepwise from 0 to 7 mM. A. globiformis won the competition for limiting amounts of ammonium when the glucose concentration in the reservoirs increased, in agreement with previously described experiments in which the flagellates were not included. The numbers of nitrifying bacteria decreased as the numbers of heterotrophic bacteria rose with increasing glucose concentrations. Critical C/N ratios, i.e., ratios between glucose and ammonium in the reservoirs at which no nitrate was found in the culture vessels, of 12.5 and 10.5 were determined at dilution rates of 0.004 and 0.01 h, respectively. Below these critical values, coexistence of the competing species was found. The numbers of nitrifying bacteria decreased more in the presence of flagellates than in their absence, presumably by selective predation on the nitrifying bacteria, either in the liquid culture or on the glass wall of the culture vessels. Despite this, the rate of nitrate production did not decrease more in the presence of flagellates than in their absence. This demonstrates that no correlation has to be expected between numbers of nitrifying bacteria and their activity and that a constant nitrification rate per cell cannot be assumed for nitrifying bacteria. Above the critical C/N ratios, low numbers of nitrifying bacteria were still found in the culture vessels, probably because of attachment of the nitrifying bacteria to the glass wall of the culture vessels. Like the numbers of heterotrophic bacteria, the numbers of flagellates increased when the glucose concentrations in the reservoirs increased. Numbers of 2 x 10 and 12 x 10 flagellates ml were found at 7 mM glucose at dilution rates of 0.004 and 0.01 h, respectively. It was concluded that the critical C/N ratios were practically unaffected by the presence of protozoa. Although nitrate production rates were equal in the presence and absence of flagellates, the numbers of nitrifying bacteria decreased more strongly in their presence. This indicates a higher activity per nitrifying cell in the presence of flagellates. PMID- 16348723 TI - Recalcitrant high-molecular-weight material, an inhibitor of microbial metabolism in river biofilms. AB - Recalcitrant high (>1,000)-molecular-weight materials (>1K materials) have been proposed to inhibit the metabolic activity of river biofilms. Evidence is presented to support the hypothesis that the mode of action is through occlusion of the surface of the biofilm by recalcitrant >1K materials. The evidence includes the following. (i) The phenomenon was particularly prominent in brown water rivers, which are rich in recalcitrant >1K material. (ii)Temporal changes in inhibition intensity were observed, which coincided with seasonal changes in the relative recalcitrance of river water >1K materials. (iii) Stores of intracellular carbon (poly-beta-hydroxyalkanoates) were progressively compromised by increasing the supply of recalcitrant >1K materials. (iv) Nontoxic synthetic analogs of recalcitrant >1K materials were also demonstrated to be capable of producing an identical inhibitory effect. Finally, it was suggested that the application of recalcitrant >1K materials may represent a novel approach to the control of deleterious microbial communities. PMID- 16348724 TI - Differentiation of gram-negative, nonfermentative bacteria isolated from biofilters on the basis of Fatty Acid composition, quinone system, and physiological reaction profiles. AB - Gram-negative, nonfermentative bacteria isolated from biofilters for off-gas treatment of animal-rendering-plant emissions were differentiated by whole-cell fatty acid analysis, quinone analysis, and numerical taxonomy based on their physiological reaction profiles. The last system consisted of 60 physiological tests and was arranged as a microtest system on microtitration plates. Based on fatty acid analyses, 31 isolates were separated into six clusters and five single member clusters. The isolates of two clusters were identified as Alcaligenes faecalis and Pseudomonas diminuta. The remaining nine clusters were characterized by their fatty acid profiles, quinone systems, and physiological reaction profiles. Clusters resulting from fatty acid analyses were compared with those resulting from physiological reaction profiles. Six clusters could be confirmed this way. The efficiency of the physiological test system was increased by the prearrangement of the isolates according to their quinone type. PMID- 16348725 TI - Evaluation of Chemicals for Restricting Colony Spreading by a Xerophilic Mold, Eurotium amstelodami, on Dichloran-18% Glycerol Agar. AB - Twenty chemicals were screened for their effectiveness in restricting colony spreading by four strains of a xerophilic mold, Eurotium amstelodami, on dichloran-18% glycerol agar. Triton X-100, Triton X-301, Tergitol NP-7, and Tergitol 15-S-3 (each at 200 mug/ml) and 1,000 mug of sodium deoxycholate, 1 mug of iprodione, 0.1 mug of propiconazole, and 0.01 mug of Maxim per ml were judged to be most effective for restricting the rate of colony spreading. PMID- 16348726 TI - Enhanced Control of Listeria monocytogenes by In Situ-Produced Pediocin during Dry Fermented Sausage Production. AB - [This corrects the article on p. 887 in vol. 58.]. PMID- 16348727 TI - Biochemical basis for whole-cell uptake kinetics: specific affinity, oligotrophic capacity, and the meaning of the michaelis constant. AB - [This corrects the article on p. 2037 in vol. 57.]. PMID- 16348728 TI - Inhibition of Settlement by Larvae of Balanus amphitrite and Ciona intestinalis by a Surface-Colonizing Marine Bacterium. AB - In an attempt to isolate bacteria with inhibitory effects against settlement by larvae of sessile invertebrates, 40 marine bacterial isolates were screened for effects against laboratory-reared barnacle larvae (Balanus amphitrite) and ascidian larvae (Ciona intestinalis). Five isolates displayed non-pH-dependent inhibitory effects against the larvae. The initial characterization of a toxic component released from an isolate, designated D2 (CCUG 26757), and its effect on laboratory-reared barnacle and ascidian larvae were studied. D2 is a facultative, anaerobic, gram-negative bacterium isolated from the surface of C. intestinalis from waters off the Swedish west coast at a depth of 10 m. Results suggest that the toxic component is released by D2 during the stationary phase. Aged biofilms were more toxic to the larvae than unaged films. The biologically active compound was in the supernatant of D2 and was heat stable and <500 Da in molecular mass. No evidence of protein or peptide moieties was found. On the basis of two phase and chromatography separations, the component is polar and neutral and contains or binds to carbohydrate moieties. Metaperiodate treatment increased toxicity; undiluted supernatant from a 24-h growth culture of D2 killed barnacle and ascidian larvae within a few hours of exposure, whereas after metaperiodate treatment, the larvae were killed in approximately 30 min. PMID- 16348729 TI - Screening of nonfilamentous bacteria for production of cutin-degrading enzymes. AB - Two hundred thirty-two nonfilamentous bacterial strains, including saprophytes, plant pathogens, and opportunistic plant and human pathogens, were screened for the ability to produce cutinases (cutin-degrading esterases). Initially, esterase activity of culture filtrates of strains grown in nutrient broth-yeast extract medium supplemented with 0.4% apple or tomato cutin was determined by a spectrophotometric assay utilizing the model substrate p-nitrophenyl butyrate. The culture filtrates of the 10 Pseudomonas aeruginosa strains tested exhibited the highest esterase activity, with values of >500 nmol/min/ml. Of these 10 strains, 3 (K799, 1499A, and DAR41352) demonstrated significant induction (10 fold or above) of esterase activity by addition of cutin to nutrient broth-yeast extract medium. The ability of culture filtrates of the three strains to cause release of apple cutin monomers was confirmed by a novel high-performance liquid chromatography technique. Monomer identification was confirmed by gas chromatography-mass spectroscopy analyses. Addition of the nonionic detergent n octylglucoside stimulated cutinase activity of culture filtrates from strains K799 and DAR41352, but not that of filtrates from strain 1499A. Time course studies in nutrient broth-yeast extract medium supplemented with apple cutin indicated maximal levels of cutinase in the culture fluids after cultures entered stationary phase. Incubation temperatures below the optimal temperature for growth (37 degrees C) led to maximal production of cutinase. PMID- 16348730 TI - Diversity among Rhizobia Effective with Robinia pseudoacacia L. AB - The diversity of rhizobia that form symbioses with roots of black locust (Robinia pseudoacacia L.), an economically important leguminous tree species, was examined by inoculating seedling root zones with samples of soil collected from the United States, Canada, and China. Bacteria were isolated from nodules, subcultured, and verified to be rhizobia. The 186 isolates varied significantly in their resistance to antibiotics and NaCl, their growth on different carbohydrates, and their effect on the pH of culture media. Most isolates showed intermediate antibiotic resistance, the capacity to use numerous carbohydrates, and a neutral to acid pH response. Isolates had greater similarity within sampling locations than among sampling locations. The isolates were grouped by using numerical taxonomy techniques, and representative strains of 37 groups were selected. The mean generation times of these isolates ranged from 3 to 9 h, and the protein profile of each of the 37 isolates was unique. Nitrogen fixation, total nitrogen accumulation, and plant growth varied significantly among black locust seedlings inoculated with the representative isolates. We conclude that great variation exists among Rhizobium spp. that nodulate black locust, and selection of strains for efficiency of the symbiotic association appears possible. PMID- 16348731 TI - Detection of Active Yeast Cells (Saccharomyces cerevisiae) in Frozen Dough Sections. AB - A new method based on fluorescence microscopy was developed to detect active yeast cells in cryosections of wheat dough. The sections were stained with 4',6 diamidino-2-phenylindole (DAPI) and counterstained with Evans blue. The active yeast cells in the sections appeared brilliant yellow and were readily distinguished from the red dough matrix. The dead cells allowed penetration of the Evans blue through the cell membrane, which interfered with the DAPI staining and caused the dead cells to blend into the red environment. The number of active yeast cells in fermenting dough sections containing different proportions of living and dead yeast cells correlated well with the gas-forming capability of the yeast in the dough but not with the results of the conventional plate count method. The new method allows the study of yeast activity not only during the different stages of frozen dough processing but also during the fermentation of doughs. PMID- 16348732 TI - Enzymes Involved in Anaerobic Polyethylene Glycol Degradation by Pelobacter venetianus and Bacteroides Strain PG1. AB - In extracts of polyethylene glycol (PEG)-grown cells of the strictly anaerobically fermenting bacterium Pelobacter venetianus, two different enzyme activities were detected, a diol dehydratase and a PEG-degrading enzyme which was characterized as a PEG acetaldehyde lyase. Both enzymes were oxygen sensitive and depended on a reductant, such as titanium citrate or sulfhydryl compounds, for optimal activity. The diol dehydratase was inhibited by various corrinoids (adenosylcobalamin, cyanocobalamin, hydroxocobalamin, and methylcobalamin) by up to 37% at a concentration of 100 muM. Changes in ionic strength and the K ion concentration had only limited effects on this enzyme activity; glycerol inhibited the enzyme by 95%. The PEG-degrading enzyme activity was stimulated by the same corrinoids by up to 80%, exhibited optimal activity in 0.75 M potassium phosphate buffer or in the presence of 4 M KCI, and was only slightly affected by glycerol. Both enzymes were located in the cytoplasmic space. Also, another PEG degrading bacterium, Bacteroides strain PG1, contained a PEG acetaldehyde lyase activity analogous to the corresponding enzyme of P. venetianus but no diol dehydratase. Our results confirm that corrinoid-influenced PEG degradation analogous to a diol dehydratase reaction is a common strategy among several different strictly anaerobic PEG-degrading bacteria. PMID- 16348733 TI - Reductive desulfurization of dibenzyldisulfide. AB - Dibenzyldisulfide was reductively degraded by a methanogenic mixed culture derived from a sewage digestor. Toluene was produced with benzyl mercaptan as an intermediate in sulfur-limited medium. Toluene production was strictly associated with biological activity; however, the reducing agent for the culture medium, Ti(III), was partially responsible for production of benzyl mercaptan. Sulfide was not detected. Additions of sodium sulfide did not inhibit toluene production. Additions of 2-bromoethane sulfonic acid prevented methanogenesis but did not adversely affect toluene yields. PMID- 16348734 TI - Production of Dihomo-gamma-Linolenic Acid by a Delta5-Desaturase-Defective Mutant of Mortierella alpina 1S-4. AB - A mutant, which has low Delta5-desaturase activity, of an arachidonic acid producing fungus, Mortierella alpina 1S-4, was shown to be a novel potent producer of dihomo-gamma-linolenic acid (DHGA). On submerged culture under optimal conditions for 6 days at 28 degrees C in a 10-liter fermentor, the mutant produced 3.2 g of DHGA per liter of culture broth (123 mg/g of dry mycelia), which accounted for 23.4% of the total mycelial fatty acids. Mycelial arachidonic acid amounted to only 19 mg/g of dry mycelia (0.5 g/liter of culture broth), which accounted for 3.7% of the total mycelial fatty acids. The other major mycelial fatty acids were palmitic acid (11.0%), stearic acid (12.8%), oleic acid (22.7%), linoleic acid (8.9%), gamma-linolenic acid (6.5%), and lignoceric acid (7.8%). More than 97 mol% of the DHGA produced was found in the triglyceride fraction irrespective of the growth temperature employed (12 to 28 degrees C). PMID- 16348735 TI - Anaerobic Growth of Thiobacillus ferrooxidans. AB - The obligately autotrophic acidophile Thiobacillus ferrooxidans was grown on elemental sulfur in anaerobic batch cultures, using ferric iron as an electron acceptor. During anaerobic growth, ferric iron present in the growth media was quantitatively reduced to ferrous iron. The doubling time in anaerobic cultures was approximately 24 h. Anaerobic growth did not occur in the absence of elemental sulfur or ferric iron. During growth, a linear relationship existed between the concentration of ferrous iron accumulated in the cultures and the cell density. The results suggest that ferric iron may be an important electron acceptor for the oxidation of sulfur compounds in acidic environments. PMID- 16348736 TI - Rapid method for analyzing bacterial behavioral responses to chemical stimuli. AB - A rapid method was developed to analyze bacterial behavioral responses to chemical stimuli. Digital image processing was used to detect the accumulation of bacteria at the mouth of a capillary containing an attractant. The accumulation of bacteria was determined from the total number of cells near the mouth of the capillary per videotape frame. This method was applied to measure the chemotactic response of Pseudomonas aeruginosa cells to serine, with results similar to those obtained by the classical capillary plating assay. The videotape method is much less time-consuming and makes it possible to assess the bacterial response to an attractant within a few minutes. PMID- 16348737 TI - The Composition and Attributes of Colletotrichum truncatum Spores Are Altered by the Nutritional Environment. AB - Previous sporulation studies with Colletotrichum truncatum NRRL 13737, a fungal pathogen of the noxious weed Sesbania exaltata, showed that the carbon-to nitrogen (CN) ratio of the conidiation medium influenced spore yield, morphology, and efficacy in inciting disease in S. exaltata. Spores produced in a medium with a CN ratio of 10:1 were more effective than were spores produced in a 30:1 or 80:1 ratio in causing disease in S. exaltata. With a basal salts medium supplemented with glucose and Casamino Acids, substrate utilization, spore production, biomass accumulation, and biomass and spore composition were compared in submerged cultures of C. truncatum grown in media with CN ratios of 80:1, 30:1, and 10:1. All cultures were sporulating by day 2, and spore concentrations in 5-day-old cultures were significantly different: 30:1 > 10:1 > 80:1. Amino acid and glucose utilization was balanced in cultures grown in media with a CN ratio of 10:1, whereas cultures grown in media with a CN ratio of 30:1 or 80:1 depleted amino acids prior to glucose. Conidia produced in media with a CN ratio of 10:1 contained significantly more protein (32% of dry weight) and less lipid (17% of dry weight) than conidia produced in media with a CN ratio of either 30:1 (15% protein, 33% lipid) or 80:1 (12% protein, 37% lipid). The higher lipid content of spores produced in media with a CN ratio of 30:1 or 80:1 was associated with the presence of increased numbers of lipid droplets. Optimization studies on conidia produced in media with CN ratios between 30:1 and 10:1 which compared yield, attributes, and efficacy in inciting disease in S. exaltata suggest that media with a CN ratio of 15:1 to 20:1 may be optimal for conidium production. PMID- 16348738 TI - Application of C Nuclear Magnetic Resonance To Elucidate the Unexpected Biosynthesis of Erythritol by Leuconostoc oenos. AB - Natural-abundance C nuclear magnetic resonance (C-NMR) revealed the production of erythritol and glycerol by nongrowing cells of Leuconostoc oenos metabolizing glucose. The ratio of erythritol to glycerol was strongly influenced by the aeration conditions of the medium. The elucidation of the metabolic pathway responsible for erythritol production was achieved by C-NMR and H-NMR spectroscopy using specifically C-labelled d-glucose. The H-NMR spectrum of the cell supernatant resulting from the metabolism of [2-C]glucose showed that only 75% of the glucose supplied was metabolized heterofermentatively and that the remaining 25% was channelled to the production of erythritol. The synthesis of this polyol resulted from the reduction of the C-4 moiety of the intermediate fructose 6-phosphate. Oxygen has an inhibitory effect on the production of erythritol by L. oenos. Preaeration of a suspension of nongrowing cells of L. oenos resulted in 30% less erythritol and in 70% more glycerol formed during the anaerobic metabolism of glucose. The anaerobic production of erythritol from glucose was also found in growing cultures of L. oenos, although to a smaller extent. PMID- 16348739 TI - Utilization of carbon substrates, electrophoretic enzyme patterns, and symbiotic performance of plasmid-cured clover rhizobia. AB - Plasmids in Rhizobium spp. are relatively large, numerous, and difficult to cure. Except for the symbiotic plasmid, little is known about their functions. The primary objective of our investigation was to obtain plasmid-cured derivatives of Rhizobium leguminosarum bv. trifolii by using a direct selection system and to determine changes in the phenotype of the cured strains. Three strains of rhizobia were utilized that contained three, four, and five plasmids. Phenotypic effects observed after curing of plasmids indicated that the plasmids were involved in the utilization of adonitol, arabinose, catechol, glycerol, inositol, lactose, malate, rhamnose, and sorbitol and also influenced motility, lipopolysaccharide production, and utilization of nitrate. Specific staining of 26 enzymes electrophoretically separated on starch gels indicated that superoxide dismutase, hexokinase, and carbamate kinase activities were affected by curing of plasmids. Curing of cryptic plasmids also influenced nodulation and growth of plants on nitrogen-deficient media. The alteration in the ability to utilize various substrates after curing of plasmids suggests that the plasmids may encode genes that contribute significantly to the saprophytic competence of rhizobia in soil. PMID- 16348740 TI - Oxidation of Nitrapyrin to 6-Chloropicolinic Acid by the Ammonia-Oxidizing Bacterium Nitrosomonas europaea. AB - Suspensions of Nitrosomonas europaea catalyzed the oxidation of the commercial nitrification inhibitor nitrapyrin [2-chloro-6-(trichloromethyl)-pyridine]. Rapid oxidation of nitrapyrin (at a concentration of 10 muM) required the concomitant oxidation of ammonia, hydroxylamine, or hydrazine. The turnover rate was highest in the presence of 10 mM ammonia (0.8 nmol of nitrapyrin per min/mg of protein). The product of the reaction was 6-chloropicolinic acid. By the use of O(2), it was shown that one of the oxygens in 6-chloropicolinic acid came from diatomic oxygen and that the other came from water. Approximately 13% of the radioactivity of [2,6-C]nitrapyrin was shown to bind to cells. Most (94%) of the latter was bound indiscriminately to membrane proteins. The nitrapyrin bound to membrane proteins may account for the observed inactivation of ammonia oxidation. PMID- 16348741 TI - Association of luminous bacteria with artificial and natural surfaces in arabian gulf seawater. AB - Luminous bacteria in seawater around the islands of Bahrain are predominantly Vibrio harveyi and have the capability to adhere to artificial fibrous surfaces. Phytoplankton did not appear to have any specific relationship with luminous bacteria, but macroalgae were shown to possess an enhanced concentration of luminous bacteria. PMID- 16348742 TI - Long-Term Dosimetry of Solar UV Radiation in Antarctica with Spores of Bacillus subtilis. AB - The main objective was to assess the influence of the seasonal stratospheric ozone depletion on the UV climate in Antarctica by using a biological test system. This method is based on the UV sensitivity of a DNA repair-deficient strain of Bacillus subtilis (TKJ 6321). In our field experiment, dried layers of B. subtilis spores on quartz discs were exposed in different seasons in an exposure box open to solar radiation at the German Antarctic Georg von Neumayer Station (70 degrees 37'S, 8 degrees 22'W). The UV-induced loss of the colony forming ability was chosen as the biological end point and taken as a measure for the absorbed biologically harmful UV radiation. Inactivation constants were calculated from the resulting dose-response curves. The results of field experiments performed in different seasons indicate a strongly season-dependent trend of the daily UV-B level. Exposures performed at extremely depleted ozone concentrations (October 1990) gave higher biologically harmful UV-B levels than expected from the calculated season-dependent trend, which was determined at normal ozone values. These values were similar to values which were measured during the Antarctic summer, indicating that the depleted ozone column thickness has an extreme influence on the biologically harmful UV climate on ground. PMID- 16348743 TI - Effect of protistan grazing on the frequency of dividing cells in bacterioplankton assemblages. AB - Grazing by phagotrophic flagellates and ciliates is a major source of mortality for bacterioplankton in both marine and freshwater systems. Recent studies have demonstrated a positive relationship between clearance rate and prey size for bacterivorous protists. We tested the idea that, by selectively grazing the larger (more actively growing or dividing) cells in a bacterial assemblage, protists control bacterial standing stock abundances by directly cropping bacterial production. Samples of estuarine water were passed through 0.8-mum-pore size filters (bacteria only) or 20-mum-mesh screens (bacteria and bacterivorous protists) and placed in dialysis tubing suspended in 7 liters of unfiltered water. Changes in total bacterial biovolume per milliliter (bacterial biomass), frequency of dividing cells (FDC), and average per cell biovolume were followed over a period of 24 h. In three experiments, the FDC increased more rapidly and attained higher values in water passed through 0.8-mum-pore-size filters (average, 5.1 to 8.9%; maximum, 15.5%) compared with FDC values in water passed through 20-mum-mesh screens (average, 2.7 to 5.3%; maximum, 6.7%). Increases in bacterial biomass per milliliter lagged behind increases in FDC by about 4 to 6 h. Grazed bacterial assemblages were characterized by lower total biomasses and smaller average cell sizes compared with those of cells in nongrazed assemblages. We conclude that bacterivorous protists control bacterial standing stock abundances partly by preferentially removing dividing cells. Selective grazing of the more actively growing cells may also explain, in part, the ability of slow growing cells to persist in bacterioplankton assemblages. PMID- 16348744 TI - Effect of starvation on induction of quinoline degradation for a subsurface bacterium in a continuous-flow column. AB - Differences in the induction response and the initial two reactions of quinoline degradation between short-term (2 days)- and long-term (60 to 80 days)-starved cells of a subsurface Pseudomonas cepacia strain were examined by using continuous-flow columns. The ability of bacteria that are indigenous to oligotrophic environments to respond to a contaminant was assessed by using long term starvation to induce a cell physiology that simulates the in situ physiology of the bacteria. With quinoline concentrations of 39 and 155 muM, long-term starved cells converted quinoline to degradation products more efficiently than did short-term-starved cells. Quinoline concentrations of 155 muM and, to a greater extent, 775 muM had an inhibitory effect on induction in long-term starved cells. However, only the length of the induction process was affected with these quinoline concentrations; degradation of quinoline at the steady state for long-term-starved cells was equal to or better than that for short-term starved cells. The induction time for short-term-starved cells did not increase progressively with increasing quinoline concentration. Experiments with starved cells are important for the development of accurate predictive models of contaminant transport in the subsurface because starvation, which induces a cell physiology that simulates the in situ physiology of many bacteria, may affect the induction process. PMID- 16348745 TI - Bradyrhizobium Populations Occur in Deep Soil under the Leguminous Tree Acacia albida. AB - Soil cores were drilled under the leguminous tree Acacia albida growing in two different ecoclimatic zones of West Africa: the Sahelian area (100 to 500 mm of annual rainfall) and the Sudano-Guinean area (1,000 to 1,500 mm of annual rainfall). Soil samples were collected at different depths from the surface down to the water table level and analyzed for the presence of rhizobia able to nodulate A. albida. In both areas, population densities of rhizobia were substantially greater near the water table than near the surface. In the Sahelian area, rhizobia were present as deep as 34 m at a concentration of 1.3 x 10/g of soil. In the Sudano-Guinean area, population densities at 0.5 to 4.5 m depth were higher than in the Sahelian area and, at several depths, comparable to that of temperate soils supporting legume crops (10 rhizobia per g of soil). Surface and deep soil isolates from all four sites were found to be slow-growing rhizobia (Bradyrhizobium sp.). The proportion of effective isolates was almost the same within surface and deep soils. PMID- 16348746 TI - Rates of microbenthic and meiobenthic bacterivory in a temperate muddy tidal flat community. AB - Rates of bacterivory in micro- and meiobenthic species were determined by an improved technique in a muddy tidal flat community in Boston Harbor, Mass. The predominant grazers of bacteria were identified, and their rates of grazing were measured in the top 1 cm of the sediment. Grazing rates were measured by a fluorescence-labeled bacteria (FLB) technique. A mixture of two Enterococcus spp. isolates and two isolates of Escherichia coli were prepared as FLB, and they were added to intact sediment cores by replacing the pore water in the upper centimeter of the core. A standard FLB procedure was modified by filtering sediment dilutions onto cellulose membrane filters and processing the filters to render them optically transparent while preserving the physical integrity of the micro- and meiobenthic organisms. Thus, it was possible, on the same microscopic field, to switch from light microscopy for identification of grazers to epifluorescence microscopy for counting FLB present in the gut contents of the same grazers. The majority of benthic organisms present in these sediments consumed FLB, but their consumption rates varied widely. Two ciliate species, a Prorodon sp. and a Chlamidodon sp., and a nematode, a Metoncholaimus sp., consumed fluorescence-labeled coliforms at the highest rates, 126 to 169 FLB per individual per h. Other ciliates and nematodes, as well as microflagellates and harpacticoid copepods, consumed fluorescence-labeled coliforms at lower rates, 1.2 to 26 FLB per individual per h. Foraminiferans and gastrotriches did not contain FLB. Some ciliate grazers discriminated between enterococci and coliforms, consuming the rod-shaped fluorescence-labeled coliforms at 74- to 155 fold-higher rates than did the coccus-shaped fluorescence-labeled enterococci. Other ciliates did not select between fluorescence-labeled enterococci and fluorescence-labeled coliforms. The high rates of bacterivory by some ciliates and nematodes indicated intensive grazing. However, at their low extant densities, the grazers consumed only a small portion of the bacterial standing stock. Major bacterial grazers, e.g., microflagellates, ciliates, and nematodes, could potentially consume, per day, only 0.2, 0.1, and 0.03%, respectively, of the bacterial standing stock (7.5 x 10 bacteria per cm). PMID- 16348747 TI - Starvation Response of the Marine Barophile CNPT-3. AB - The psychrophilic marine barophile CNPT-3 underwent a starvation-survival response similar to that reported for the marine bacteria Ant-300, DW1, and S-14. The number of culturable cells increased initially and then decreased gradually over a 24-day starvation period, with corresponding decreases in total cell number and direct viability count. A significant reduction in cell size and biovolume accompanied these changes. Starved cells demonstrated a greater tendency to attach at the in situ pressure (400 atm; ca. 40.5 MPa) and temperature (5 degrees C) than at 1 atm (ca. 101 kPa), and the extent of attachment increased with increasing duration of starvation. The membrane fatty acid profile of the marine barophile CNPT-3 was studied as the cells were subjected to starvation conditions. A 37.5% increase in saturated fatty acids was observed during the first 8 days of starvation, with a concomitant decrease in unsaturated fatty acids. There was also an increase in the amount of short-chain (10,000)- and low (<1,000)-molecular-weight (MW) components and sometimes contained intermediate-MW components as well. In general, bacterial turnover rates of the low-MW components were fairly rapid, whereas the high-MW components were utilized slowly or not at all. In the humic lake, the intermediate-MW components accounted for a large proportion of the net PDOC and were subject to rapid bacterial utilization. This fraction probably consisted almost entirely of polysaccharides of ca. 6,000 MW. Amino acids and peptides, other organic acids, and carbohydrates could all be quantitatively important parts of the low-MW PDOC that was utilized by the heterotrophic bacteria, but the relative contributions of these fractions differed widely. It was concluded that, generally, low-MW components of PDOC are quantitatively much more important to the bacteria than are high-MW components, that PDOC released from phytoplankton does not contain substances of quantitative importance as bacterial substrates in all situations, and that high-MW components of PDOC probably contribute to the buildup of refractory, high-MW dissolved organic carbon in pelagic environments. PMID- 16348768 TI - Molecular monitoring of wine fermentations conducted by active dry yeast strains. AB - A simple and rapid method of yeast strain characterization based on mitochondrial DNA restriction analysis was applied to the control of wine fermentations conducted by active dry yeast strains. This molecular approach allows us to understand several important aspects of this process, such as the role of the active dry yeast strain and that of the natural Saccharomyces cerevisiae flora during vinification. In this paper, we demonstrate that the inoculated strain is really responsible for the fermentation but does not suppress significant development of natural strains during the first stages. During this early period, natural strains could have important effects on wine flavor. PMID- 16348769 TI - Influence of solid surface, adhesive ability, and inoculum size on bacterial colonization in microcosm studies. AB - Microcosm studies were performed to evaluate the effect of solid surfaces, bacterial adhesive ability, and inoculum size on colonization success and persistence of Pseudomonas fluorescens and Xanthomonas maltophilia, each with a Tn5 insertion that conferred resistance to kanamycin and streptomycin. Two types of microcosms were used: (i) a simple system that was colonized by Aeromonas hydrophila and a coryneform and (ii) a complex system produced from lake water enrichment cultures. Simple microcosms contained 100 ml of peptone- and yeast extract-supplemented artificial lake water or 60 ml of peptone- and yeast extract supplemented artificial lake water with 70 g of 3-mm glass beads. Complex microcosms contained 100 ml of lake water with no nutrient additions or 100 ml of lake water with 70 g of glass beads. The microcosms were incubated for 35 days at 20 degrees C. In lake water enrichment microcosms, the presence of beads increased the abilities of P. fluorescens or X. maltophilia to colonize, but their numbers decreased with time in microcosms both with and without beads. The adhesiveness of the bacteria, measured in an in vitro assay, did not relate to colonization success. In simple microcosms, the inoculum size (10, 10, or 10) of P. fluorescens did not influence colonization success. However, in complex microcosms, an inoculum of 10 cells was insufficient to ensure colonization by P. fluorescens, while 10 cells resulted in colonization of liquid and beads. Simple microcosm studies, utilizing only a few species, were poor models for complex natural systems. In complex enrichment systems, colonization of surfaces resulted in higher numbers of organisms but did not noticeably promote persistence. Adhesiveness of a particular organism may be a relatively minor factor influencing its ability to colonize solid surfaces in complex natural environments. PMID- 16348770 TI - Ice Nucleation Activity in Fusarium acuminatum and Fusarium avenaceum. AB - Twenty fungal genera, including 14 Fusarium species, were examined for ice nucleation activity at -5.0 degrees C, and this activity was found only in Fusarium acuminatum and Fusarium avenaceum. This characteristic is unique to these two species. Ice nucleation activity of F. avenaceum was compared with ice nucleation activity of a Pseudomonas sp. strain. Cumulative nucleus spectra are similar for both microorganisms, while the maximum temperatures of ice nucleation were -2.5 degrees C for F. avenaceum and -1.0 degrees C for the bacteria. Ice nucleation activity of F. avenaceum was stable at pH levels from 1 to 13 and tolerated temperature treatments up to 60 degrees C, suggesting that these ice nuclei are more similar to lichen ice nuclei than to bacterial ones. Ice nuclei of F. avenaceum, unlike bacterial ice nuclei, pass through a 0.22-mum-pore-size filter. Fusarial nuclei share some characteristics with the so-called leaf derived nuclei with which they might be identified: they are cell free and stable up to 60 degrees C, and they are found in the same kinds of environment. Highly stable ice nuclei produced by fast-growing microorganisms have potential applications in biotechnology. This is the first report of ice nucleation activity in free-living fungi. PMID- 16348771 TI - Evaluation of methyl fluoride and dimethyl ether as inhibitors of aerobic methane oxidation. AB - Methyl fluoride (MF) and dimethyl ether (DME) were effective inhibitors of aerobic methanotrophy in a variety of soils. MF and DME blocked consumption of CH(4) as well as the oxidation of CH(4) to CO(2), but neither MF nor DME affected the oxidation of [C]methanol or [C]formate to CO(2). Cooxidation of ethane and propane by methane-oxidizing soils was also inhibited by MF. Nitrification (ammonia oxidation) in soils was inhibited by both MF and DME. Production of N(2)O via nitrification was inhibited by MF; however, MF did not affect N(2)O production associated with denitrification. Methanogenesis was partially inhibited by MF but not by DME. Methane oxidation was approximately 100-fold more sensitive to MF than was methanogenesis, indicating that an optimum concentration could be employed to selectively block methanotrophy. MF inhibited methane oxidation by cell suspensions of Methylococcus capsulatus; however, DME was a much less effective inhibitor. PMID- 16348773 TI - Assessment of Pediococcus acidilactici as a Potential Silage Inoculant. AB - Eighteen Pediococcus strains were screened for their potential as silage inoculants. Pediococcus acidilactici G24 was found to be the most suitable, exhibiting a short lag phase on both glucose and fructose, a rapid rate of acid production, a high sugar-to-lactate conversion efficiency, no detectable breakdown of proteins or lactic acid, and the ability to grow within a broad range of pH and temperature. When tested in laboratory silos using grass with a water-soluble carbohydrate content of 24 g/kg of aqueous extract, P. acidilactici G24 stimulated the natural Lactobacillus plantarum population and accelerated the rates of lactic acid production and pH decrease. After 6 days of fermentation, the inoculated silage exhibited a 12% decrease in ammonia nitrogen and an 11% increase in crude protein levels compared with uninoculated controls. The use of an L. plantarum inoculant at a rate of 10 bacteria per g of grass in conjunction with P. acidilactici G24 produced no additional beneficial effect. Inoculation of grass with a water-soluble carbohydrate level of 8 g/kg of aqueous extract with P. acidilactici G24 led to no acceleration in the rate of L. plantarum growth or pH decrease. However, after 7 days of fermentation the inoculated silage had a 14% lower ammonia nitrogen protein content than did uninoculated controls. The results suggest that P. acidilactici G24 may be useful as a silage inoculant for crops with a sufficiently high water-soluble carbohydrate level. PMID- 16348772 TI - Persistence of free plasmid DNA in soil monitored by various methods, including a transformation assay. AB - The persistence and stability of free plasmid pUC8-ISP DNA introduced into 10-g samples of various soils and kept at 23 degrees C were monitored over a period of 60 days. The soils were sampled at a plant science farm and included a loamy sand soil (no. 1), a clay soil (no. 2), and a silty clay soil (no. 3). Four different methods allowed monitoring of (i) the production of acid-soluble radioactive material from [H]thymidine-labeled plasmid DNA, (ii) the decrease of hybridizing nucleotide sequences in slot blot analysis, (iii) the loss of plasmid integrity measured by Southern hybridization, and (iv) the decay of the biological activity as determined by transformation of Ca-treated Escherichia coli cells with the DNA extracted from soil. Acid-soluble material was not produced within the first 24 h but then increased to 45% (soil no. 1), 27% (soil no. 2), and 77% (soil no. 3) until the end of incubation. A quite parallel loss of material giving a slot blot hybridization signal was observed. Southern hybridization indicated that after 1 h in the soils, plasmid DNA was mostly in the form of circular and full-length linear molecules but that, depending on the soil type, after 2 to 5 days full length plasmid molecules were hardly detectable. The transforming activity of plasmid DNA reextracted from the soils followed inactivation curves over 2 to 4 orders of magnitude and dropped below the detection limit after 10 days. The inactivation was slower in soil no. 2 (28.2-h half-life time of the transforming activity of a plasmid molecule) than in soils no. 3 (15.1 h) and no. 1 (9.1 h). The studies provide data on the persistence of free DNA molecules in natural bacterial soil habitats. The data suggest that plasmid DNA may persist long enough to be available for uptake by competent recipient cells in situ. PMID- 16348774 TI - Characteristics of a hydrated, alginate-based delivery system for cultivation of the button mushroom. AB - The production of the button mushroom Agaricus bisporus with mycelium-colonized alginate pellets as an inoculant of the growing medium was investigated. Pellets having an irregular surface and porous internal structure were prepared by complexing a mixture of 1% sodium alginate, 2 to 6% vermiculite, 2% hygramer, and various concentrations of Nutrisoy (soy protein) with calcium chloride. The porous structure allowed the pellets to be formed septically and then inoculated and colonized with the fungus following sterilization. By using an enzyme-linked immunosorbent assay (ELISA) to estimate fungal biomass, the matrix components of the pellet were found to be of no nutritive value to A. bisporus. Pellets amended with Nutrisoy at a concentration of 0.5 to 8% supported extensive mycelial growth, as determined by significantly increased ELISA values, with a concentration of 4% being optimal and higher concentrations proving inhibitory. The addition of hydrated, mycelium-invaded pellets to the compost or casing layer supported the thorough colonization of the growing substrate and culminated in the formation of mushrooms that showed normal development and typical morphology. Yields and sizes of mushrooms were comparable from composts seeded with either colonized pellets or cereal grain spawn. Similarly, amending the casing layer with pelletized-mycelium-colonized compost resulted in a 2- to 3-day-earlier and more-synchronous emergence of mushrooms than with untreated casing. This technology shows the greatest potential as a pathogen-free inoculant of the casing layer in the commercial cultivation of mushrooms. PMID- 16348775 TI - Lignin Peroxidase Activity Is Not Important in Biological Bleaching and Delignification of Unbleached Kraft Pulp by Trametes versicolor. AB - The discovery in 1983 of fungal lignin peroxidases able to catalyze the oxidation of nonphenolic aromatic lignin model compounds and release some CO(2) from lignin has been seen as a major advance in understanding how fungi degrade lignin. Recently, the fungus Trametes versicolor was shown to be capable of substantial decolorization and delignification of unbleached industrial kraft pulps over 2 to 5 days. The role, if any, of lignin peroxidase in this biobleaching was therefore examined. Several different assays indicated that T. versicolor can produce and secrete peroxidase proteins, but only under certain culture conditions. However, work employing a new lignin peroxidase inhibitor (metavanadate ions) and a new lignin peroxidase assay using the dye azure B indicated that secreted lignin peroxidases do not play a role in the T. versicolor pulp-bleaching system. Oxidative activity capable of degrading 2-keto-4-methiolbutyric acid (KMB) appeared unique to ligninolytic fungi and always accompanied pulp biobleaching. PMID- 16348776 TI - Dispersion of Small Ceramic Particles (Al(2)O(3)) with Azotobacter vinelandii. AB - The high surface charge of small ceramic particles such as alumina particles prevents them from dispersing evenly in aqueous suspensions and forming high density compacts. However, suspensions of 400-nm-diameter alumina particles treated with alginate from the bacterium Azotobacter vinelandii were well dispersed. The alginate bound firmly to the particle surface and could not be removed by repeated washing with distilled water (2.82 mg of the bacterial alginate adsorbed to 1 g of the alumina particles). Furthermore, A. vinelandii grew and produced alginate in the presence of up to 15% (vol/vol) alumina particles. These results suggest that an in situ process using this bacterium to coat ceramic particles with alginate might be developed. In in situ processing experiments, the particle-packing densities were significantly increased and the viscosities of 5 and 10% (vol/vol) suspensions were reduced 4- and 60-fold, respectively, over those of controls. The bacteria were readily removed from the alumina particles by washing. PMID- 16348777 TI - Effect of Endosulfan on Azospirillum lipoferum Growth, Morphology, Nitrogenase Activity, and Protein Binding. AB - The organochlorine Thiodan CE inhibited growth and nitrogenase activity of Azospirillum lipoferum. The active ingredient, Endosulfan, was nonspecifically bound to proteins and mainly adsorbed to the cell envelope with small amounts transported into cytosol. The involvement of the external membrane and cyst formation in protection against hazardous substances is discussed. PMID- 16348778 TI - Occurrence of fumonisin in forage grass in new zealand. AB - Fumonisin B(1) (FB(1)) was isolated from samples of forage grass originating in paddocks associated with an idiopathic disease of Canadian wapiti and wapiti-red deer hybrids characterized by "ill thrift" and liver dysfunction. Four of 40 samples contained 1, 3, 6, and 9 ppm (micrograms per gram) of FB(1) and 4, 0.5, 2, and 0.5 ppm, respectively, of the methyl ester of FB(1). Analyses were done by ion spray mass spectrometry and confirmed by both fast atom bombardment (solids probe) and mass spectral analysis by electron impact ionization of the trifluoroacetate derivative of the base hydrolyzed product (pentolamine) of FB(1). This article contains the first report of the presence of fumonisin B(1) in grass. PMID- 16348779 TI - Effects of Grazing by Flagellates on Competition for Ammonium between Nitrifying and Heterotrophic Bacteria in Chemostats. AB - [This corrects the article on p. 1964 in vol. 58.]. PMID- 16348780 TI - A Hydrogen-Oxidizing, Fe(III)-Reducing Microorganism from the Great Bay Estuary, New Hampshire. AB - A dissimilatory Fe(III)- and Mn(IV)-reducing bacterium was isolated from bottom sediments of the Great Bay estuary, New Hampshire. The isolate was a facultatively anaerobic gram-negative rod which did not appear to fit into any previously described genus. It was temporarily designated strain BrY. BrY grew anaerobically in a defined medium with hydrogen or lactate as the electron donor and Fe(III) as the electron acceptor. BrY required citrate, fumarate, or malate as a carbon source for growth on H(2) and Fe(III). With Fe(III) as the sole electron acceptor, BrY metabolized hydrogen to a minimum threshold at least 60 fold lower than the threshold reported for pure cultures of sulfate reducers. This finding supports the hypothesis that when Fe(III) is available, Fe(III) reducers can outcompete sulfate reducers for electron donors. Lactate was incompletely oxidized to acetate and carbon dioxide with Fe(III) as the electron acceptor. Lactate oxidation was also coupled to the reduction of Mn(IV), U(VI), fumarate, thiosulfate, or trimethylamine n-oxide under anaerobic conditions. BrY provides a model for how enzymatic metal reduction by respiratory metal-reducing microorganisms has the potential to contribute to the mobilization of iron and trace metals and to the immobilization of uranium in sediments of Great Bay Estuary. PMID- 16348781 TI - Solubilization and mineralization of lignin by white rot fungi. AB - The white rot fungi Lentinula edodes, Phanerochaete chrysosporium, Pleurotus sajor-caju, Flammulina velutipes, and Schizophyllum commune were grown in liquid media containing C-lignin-labelled wood, and the formation of water-soluble C labelled products and CO(2), the growth of the fungi, and the activities of extracellular lignin peroxidase, manganese peroxidase, and laccase were measured. Conditions that affect the rate of lignin degradation were imposed, and both long term (0- to 16-day) and short-term (0- to 72-h) effects on the production of the two types of product and on the activities of the enzymes were monitored. The production of CO(2)-labelled products from the aqueous ones was also investigated. The short-term studies showed that the different conditions had different effects on the production of the two products and on the activities of the enzymes. Nitrogen sources inhibited the production of both products by all species when differences in growth could be discounted. Medium pH and manganese affected lignin degradation by the different species differently. With P. chrysosporium, the results were consistent, with lignin peroxidase playing a role in lignin solubilization and manganese peroxidase being important in subsequent CO(2) production. PMID- 16348783 TI - Stability and Specificity of the Cell Wall-Associated Proteinase from Lactococcus lactis subsp. cremoris H2 Released by Treatment with Lysozyme in the Presence of Calcium Ions. AB - The cell wall-associated proteinase from Lactococcus lactis subsp. cremoris H2 (isolate number 4409) was released from the cells by treatment with lysozyme, even in the presence of 50 mM calcium chloride. Cell lysis during lysozyme treatment was minimal. The proteinase activity released by lysozyme treatment fractionated on ion-exchange chromatography as three main forms, the molecular masses of which were determined by gel exclusion chromatography and polyacrylamide gel electrophoresis. Two of the enzyme forms released, 137 and 145 kDa, were the same as those released by incubation of cells in calcium-free phosphate buffer. In the presence of calcium, lysozyme treatment also resulted in the release of a 180-kDa enzyme molecule. The total proteinase activity released by lysozyme treatment (in the presence or absence of calcium) was not only greater than that released by phosphate buffer but was also greater than that initially detectable on the surface of whole cells, suggesting an unmasking of enzyme on the cell surface. The presence of calcium during release treatment resulted in increased stability of the crude enzyme preparations. For the proteinase preparation released by using lysozyme with 50 mM CaCl(2), the half life of proteinase activity at 37 degrees C was 39 h, compared with 0.22 h for the calcium-free phosphate buffer-released preparation. In all cases, maximum stability was observed at pH 5.5. Comparison of beta-casein hydrolysis by the three forms of the enzyme showed that the products of short-term (5- to 30-min) digestions were very similar, although subtle differences were detected with the 180-kDa form. PMID- 16348782 TI - Biochemical and Electron Microscopic Studies of the Streptomyces reticuli Cellulase (Avicelase) in Its Mycelium-Associated and Extracellular Forms. AB - Streptomyces reticuli is able to grow efficiently with crystalline cellulose (Avicel) as the sole carbon source. Cultivation in the presence of the nonionic detergent Tween 80 at a concentration of 0.1% led to a 10-fold increase in extracellular cellulolytic activity. Under these conditions, one single 82-kDa cellulase (Avicelase) capable of degrading crystalline and soluble cellulose as well as cellodextrins and p-nitrophenylcellobioside was purified to apparent homogeneity by a procedure which consisted of two consecutive anion-exchange chromatographies followed by chromatofocusing. Aggregation, which was a major problem during protein purification, could be avoided by including Triton X-100 at a concentration of 0.1% in every chromatographic step. The Avicelase was identified in extracellular and mycelium-associated forms, the latter of which could be released efficiently by nonionic detergents. In addition, a 42-kDa truncated form retaining cellulolytic activity was identified which had been generated from the 82-kDa enzyme by a protease. Antibodies raised against the mycelium-associated Avicelase reacted with the 42-kDa derivative and the extracellular form. The mycelial association of the enzyme was confirmed by immunofluorescence and immunoelectron microscopies. PMID- 16348784 TI - Purification and Characterization of a Keratinase from a Feather-Degrading Bacillus licheniformis Strain. AB - A keratinase was isolated from the culture medium of feather-degrading Bacillus licheniformis PWD-1 by use of an assay of the hydrolysis of azokeratin. Membrane ultrafiltration and carboxymethyl cellulose ion-exchange and Sephadex G-75 gel chromatographies were used to purify the enzyme. The specific activity of the purified keratinase relative to that in the original medium was approximately 70 fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and Sephadex G-75 chromatography indicated that the purified keratinase is monomeric and has a molecular mass of 33 kDa. The optimum pH and the pI were determined to be 7.5 and 7.25, respectively. Under standard assay conditions, the apparent temperature optimum was 50 degrees C. The enzyme is stable when stored at -20 degrees C. The purified keratinase hydrolyzes a broad range of substrates and displays higher proteolytic activity than most proteases. In practical applications, keratinase is a useful enzyme for promoting the hydrolysis of feather keratin and improving the digestibility of feather meal. PMID- 16348786 TI - Electrical energy changes conductivity and determines optimal electrotransformation frequency in gram-negative bacteria. AB - In many bacterial electrotransformation protocols, pulse time is related to the time constant for a capacitor discharging across a sample of fixed resistance. Using an electroporator which controls pulse time independently of the capacitor time constant, we found that the resistance of bacterial suspensions fluctuates widely during capacitor discharge. With three gram-negative species of bacteria, electrotransformation frequency and survival could be more simply related to the electrical energy delivered in each pulse than to component parameters, such as initial field strength, capacitance, and pulse time. In each case, the number of transformants per survivor increased exponentially and leveled off when more than 0.5 to 1.0 J of electrical energy was delivered. An inverse log-linear relationship between survival and energy delivered was also observed for all three species. PMID- 16348785 TI - Controlled Integration into the Lactococcus Chromosome of the pCI829-Encoded Abortive Infection Gene from Lactococcus lactis subsp. lactis UC811. AB - The phage insensitivity gene of lactococcal plasmid pCI829 which encodes an abortive infection defense mechanism (Abi) was inserted into the Lactococcus lactis subsp. lactis CH919 chromosome by utilizing the integration plasmid pCI194, which contains 4.2 kb of homology with the conjugative transposon Tn919. Chloramphenicol-resistant transformants expressed phage insensitivity to the prolate-headed phage c2 and the small isometric-headed phage 712, and hybridization analysis indicated that transformants contained pCI194 integrated in single copy. The level of phage insensitivity expressed by the transformants was reduced from that observed when the abi gene was located on a replicating plasmid, as determined by plaque assay and burst size analysis. Amplification of the integrated structure after growth in increased concentrations of chloramphenicol resulted in an increase in the expression of phage insensitivity. Hybridization analysis revealed that while pCI194 was stably maintained in an integrated state over 100 generations in the absence of selective pressure, the ability to express phage insensitivity was lost. Hybridization analysis also revealed that DNA flanking the abi gene contains homology to the CH919 chromosome. PMID- 16348787 TI - Competition for Ammonium between Nitrifying and Heterotrophic Bacteria in Continuously Percolated Soil Columns. AB - Although the absence of nitrate formation in grassland soils rich in organic matter has often been reported, low numbers of nitrifying bacteria are still found in these soils. To obtain more insight into these observations, we studied the competition for limiting amounts of ammonium between the chemolithotrophic ammonium-oxidizing species Nitrosomonas europaea and the heterotrophic species Arthrobacter globiformis in the presence of Nitrobacter winogradskyi with soil columns containing calcareous sandy soil. The soil columns were percolated continuously at a dilution rate of 0.007 h, based on liquid volumes, with medium containing 5 mM ammonium and different amounts of glucose ranging from 0 to 12 mM.A. globiformis was the most competitive organism for limiting amounts of ammonium. The numbers of N. europaea and N. winogradskyi cells were lower at higher glucose concentrations, and the potential ammonium-oxidizing activities in the uppermost 3 cm of the soil columns were nonexistent when at least 10 mM glucose was present in the reservoir, although 10 nitrifying cells per g of dry soil were still present. This result demonstrated that there was no correlation between the numbers of nitrifying bacteria and their activities. The numbers and activities of N. winogradskyi cells decreased less than those of N. europaea cells in all layers of the soil columns, probably because of heterotrophic growth of the nitrite-oxidizing bacteria on organic substrates excreted by the heterotrophic bacteria or because of nitrate reduction at reduced oxygen concentrations by the nitrite-oxidizing bacteria. Our conclusion was that the nitrifying bacteria were less competitive than the heterotrophic bacteria for ammonium in soil columns but that they survived as viable inactive cells. Inactive nitrifying bacteria may also be found in the rhizosphere of grassland plants, which is rich in organic carbon. They are possibly reactivated during periods of net mineralization. PMID- 16348788 TI - Carbon Monoxide, Hydrogen, and Formate Metabolism during Methanogenesis from Acetate by Thermophilic Cultures of Methanosarcina and Methanothrix Strains. AB - CO and H(2) have been implicated in methanogenesis from acetate, but it is unclear whether they are directly involved in methanogenesis or electron transfer in acetotrophic methanogens. We compared metabolism of H(2), CO, and formate by cultures of the thermophilic acetotrophic methanogens Methanosarcina thermophila TM-1 and Methanothrix sp. strain CALS-1. M. thermophila accumulated H(2) to partial pressures of 40 to 70 Pa (1 Pa = 0.987 x 10 atm), as has been previously reported for this and other Methanosarcina cultures. In contrast, Methanothrix sp. strain CALS-1 accumulated H(2) to maximum partial pressures near 1 Pa. Growing cultures of Methanothrix sp. strain CALS-1 initially accumulated CO, which reached partial pressures near 0.6 Pa (some CO came from the rubber stopper) during the middle of methanogenesis; this was followed by a decrease in CO partial pressures to less than 0.01 Pa by the end of methanogenesis. Accumulation or consumption of CO by cultures of M. thermophila growing on acetate was not detected. Late-exponential-phase cultures of Methanothrix sp. strain CALS-1, in which the CO partial pressure was decreased by flushing with N(2)-CO(2), accumulated CO to 0.16 Pa, whereas cultures to which ca. 0.5 Pa of CO was added consumed CO until it reached this partial pressure. Cyanide (1 mM) blocked CO consumption but not production. High partial pressures of H(2) (40 kPa) inhibited methanogenesis from acetate by M. thermophila but not by Methanothrix sp. strain CALS-1, and 2 kPa of CO was not inhibitory to M. thermophila but was inhibitory to Methanothrix sp. strain CALS-1. Levels of CO dehydrogenase, hydrogenase, and formate dehydrogenase in Methanothrix sp. strain CALS-1 were 9.1, 0.045, and 5.8 mumol of viologen reduced min mg of protein. These results suggest that CO plays a role in Methanothrix sp. strain CALS-1 similar to that of H(2) in M. thermophila and are consistent with the conclusion that CO is an intermediate in a catabolic or anabolic pathway in Methanothrix sp. strain CALS-1; however, they could also be explained by passive equilibration of CO with a metabolic intermediate. PMID- 16348789 TI - Degradation of Extracellular beta-(1,3)(1,6)-d-Glucan by Botrytis cinerea. AB - During growth on glucose, Botrytis cinerea produced extracellular beta-(1,3)(1,6) d-glucan (cinerean), which formed an adhering capsule and slime. After glucose was exhausted from the medium, cinereanase activity increased from <0.4 to 30 U/liter, effecting a striking loss in the viscosity of the culture. Cinerean was cleaved into glucose and gentiobiose. Gentiobiose was then hydrolyzed to glucose. While cinereanase activity was strongest in the culture supernatant, gentiobiase activity was located mainly in the cell wall fraction. The addition of extra glucose or cycloheximide prevented the cinerean degradation caused by an effect on cinereanase formation. Cinerean degradation was accompanied by microconidiation and sclerotium formation. B. cinerea was found to grow on cinerean with the latter as its single carbon and energy source. In this case, cinerean degradation occurred during hyphal growth, and no microconidiation or sclerotium formation was observed. Growth experiments with various carbon sources indicated that cinerean had a positive effect on the formation of cinerean degrading enzymes. PMID- 16348790 TI - Lysogeny in Bradyrhizobium japonicum and Its Effect on Soybean Nodulation. AB - Rhizobiophage V, isolated from soil in the vicinity of soybean roots, was strongly lytic on Bradyrhizobium japonicum 123B (USDA 123) but only mildly lytic on strain L4-4, a chemically induced small-colony mutant of 123. Numerous bacteriophage-resistant variants were isolated from L4-4 infected with phage V; two were studied in detail and shown to be lysogenic. The two, L4-4 (V5) and L4-4 (V12), are the first reported examples of temperate-phage infection in B. japonicum. Phage V and its derivative phages V5 and V12 were closely related on the basis of common sensitivity to 0.01 M sodium citrate and phage V antiserum, phage immunity tests, and apparently identical morphology when examined by electron microscopy. However, the three phages differed in host range and in virulence. Lysogens L4-4 (V5) and L4-4 (V12) were immune to infection by phages V and V5 but not to infection by V12. Southern hybridization analysis confirmed the incorporation of phage V into the genomes of strains L4-4(V5) and L4-4(V12) and also demonstrated the incorporation of phage V into the genome of a phage V resistant derivative of USDA 123 designated 123 (V2). None of the three lysogens, L4-4(V5), L4-4(V12), or 123B(V2), was able to nodulate soybean plants. However, Southern hybridization profile data indicated that phage V had not incorporated into any of the known B. japonicum nodulation genes. PMID- 16348791 TI - Comparison of Identification Systems for Classification of Bacteria Isolated from Water and Endolithic Habitats within the Deep Subsurface. AB - One water and three rock samples were taken from a mined tunnel system, U12n, in Rainier Mesa at the Nevada Test Site. Endolithic microorganisms were cultured from ashfall tuff, which was crushed and made into slurries with a formulation of artificial pore water, on R2A agar plates. Microbial counts ranged from 10 to 10 viable cells per g (dry weight) of rock sampled. The cultured water sample yielded 10 viable cells per ml. Many of the isolates were very small (<1 mum) when viewed in the rock matrix and remained small even when cultured. Most were gram-negative rods. Individual isolates were profiled by API-NFT strip number, antibiotic and metal resistance patterns, and colony and cellular morphologies. Three identification systems, API-NFT strips, BIOLOG, and MIDI, were compared. Each system identified only a small percentage of the total isolates, and in only seven cases were the isolates identified the same way by more than one system. The same genus was identified in three of these cases, but different species were indicated. The genus Pseudomonas was the most commonly identified. The isolate profiles and the three identification systems demonstrated that water isolates were considerably different from endolithic isolates. PMID- 16348792 TI - Enumeration and Characterization of Bacterial Colonists of a Submersed Aquatic Plant, Eurasian Watermilfoil (Myriophyllum spicatum L.). AB - A simple procedure for enumerating and grouping the bacterial colonists of Eurasian watermilfoil (Myriophyllum spicatum L.) is described. Colony characteristics of bacteria associated with M. spicatum were better defined and more stable on nutrient-poor, diluted nutrient broth agar than on high-nutrient media. Acinetobacter, Cytophaga, Flavobacterium, Moraxella, Pseudomonas and/or Alcaligenes, and Vibrio/Aeromonas spp., as well as two highly fastidious unidentified bacterial groups (gram-negative rods and gram-negative cocci), were associated with cultured watermilfoil during January, February, May, June, July, and August 1988. In Lake Wingra (Madison, Wis.), Micrococcus spp. and enterobacters were also associated with Eurasian watermilfoil during July, August, and October 1987. PMID- 16348793 TI - Riboflavin Production during Growth of Micrococcus luteus on Pyridine. AB - Micrococcus luteus produced 29 muM riboflavin during growth on 6.5 mM pyridine but not during growth on other substrates. On the basis of the results of radiolabelling studies, riboflavin was not directly synthesized from pyridine. Pyridine may interfere with riboflavin biosynthesis or elicit a general stress response in M. luteus. PMID- 16348794 TI - A Lactobacillus helveticus-Specific DNA Probe Detects Restriction Fragment Length Polymorphisms in This Species. AB - A cloned 2-kb EcoRI fragment (fragment f) from a 34-kb plasmid of Lactobacillus helveticus CNRZ 1094 was shown by dot blot to specifically hybridize to total DNAs of 75 L. helveticus strains. No hybridization was found with L. acidophilus, L. crispatus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. gasseri, or L. intestinalis strains. When Southern blots of EcoRI digests of L. helveticus strains were probed with fragment f, these strains displayed restriction fragment length polymorphisms on the basis of which they could be grouped into several clusters. PMID- 16348795 TI - Stable defined substrate for turbidimetric assay of endoxylanases. AB - A stable xylan suspension was prepared and characterized. Hydrolysis of the particles converts them into soluble fragments, thereby lowering the turbidity of the suspension. The small volume of the assay mixture, the short incubation time required, and the simplicity of the procedure permit the rapid analysis of many samples. Furthermore, the procedure can be used to assay xylanase activities in the presence of other reducing materials and is also useful for monitoring low level xylanase activities. PMID- 16348796 TI - Applications of a colorimetric plate assay for soluble methane monooxygenase activity. AB - [This corrects the article on p. 2235 in vol. 58.]. PMID- 16348797 TI - Scanning electron and light microscopic study of microbial succession on bethlehem st. Nectaire cheese. AB - St. Nectaire cheese is a semisoft cheese of French origin that, along with Brie and Camembert cheeses, belongs to the class of surface mold-ripened cheese. The surface microorganisms that develop on the cheese rind during ripening impart a distinctive aroma and flavor to this class of cheese. We have documented the sequential appearance of microorganisms on the cheese rind and in the curd over a 60-day ripening period. Scanning electron microscopy was used to visualize the development of surface fungi and bacteria. Light microscopy of stained paraffin sections was used to study cross sections through the rind. We also monitored the development of bacterial and yeast populations in and the pH of the curd and rind. The earliest stage of ripening (0 to 2 days) is dominated by the lactic acid bacterium Streptococcus cremoris and multilateral budding yeasts, primarily Debaryomyces and Torulopsis species. Geotrichum candidum follows closely, and then zygomycetes of the genus Mucor develop at day 4 of ripening. At day 20, the deuteromycete Trichothecium roseum appears. From day 20 until the end of the ripening process, coryneforms of the genera Brevibacterium and Arthrobacter can be seen near the surface of the cheese rind among fungal hyphae and yeast cells. PMID- 16348798 TI - Xylanase Activity of Phanerochaete chrysosporium. AB - Xylan-degrading enzymes were induced when Phanerochaete chrysosporium was grown at 30 degrees C in shake flask media containing xylan, Avicel PH 102, or ground corn stalks. The highest xylanase activity was produced in the corn stalk medium, while the xylan-based fermentation resulted in the lowest induction. Analytical and preparative isoelectric focusing were used to characterize xylanase multienzyme components. Preparative focusing was performed only with the cultures grown on Avicel and corn stalk. Of over 30 protein bands separated by analytical focusing from the Avicel and corn stalk media, three main groups (I, II, and III) of about five isoenzymes each showed xylanase activity when a zymogram technique with a xylan overlay was used. Enzyme assays revealed the presence of 1,4-beta endoxylanase and arabinofuranosidase activities in all three isoenzyme groups separated by preparative isoelectric focusing. beta-Xylosidase activity appeared in the first peak and also as an independent peak between peaks II and III. Denatured molecular masses for the three isoenzyme groups were found to be between 18 and 90 kDa, and pI values were in the range of 4.2 to 6.0. beta Xylosidase has an apparent molecular mass of 20, 30, and 90 kDa (peak I) and 18 and 45 kDa (independent peak), indicating a trimer and dimer structure, respectively, with pI values of 4.2 and 5.78, respectively. Three more minor xylanase groups were produced on corn stalk medium: a double peak in the acidic range (pI 6.25 to 6.65 and 6.65 to 7.12) and two minor peaks in the alkaline range (pI 8.09 to 8.29 and 9.28 to 9.48, respectively). The profile of xylanases separated by isoelectric focusing (zymogram) of culture filtrate from cells grown on corn stalk media was more complex than that of culture supernatants from cells grown on cellulose. The pH optima of the three major xylanase groups are in the range of pH 4 to 5.5. PMID- 16348799 TI - Comparative Physiological Studies on Hyperthermophilic Archaea Isolated from Deep Sea Hot Vents with Emphasis on Pyrococcus Strain GB-D. AB - Three new sulfur- or non-sulfur-dependent archaeal isolates, including a Pyrococcus strain, from Guaymas Basin hydrothermal vents (Gulf of California; depth, 2,010 m) were characterized and physiologically compared with four known hyperthermophiles, previously isolated from other vent sites, with an emphasis on growth and survival under the conditions particular to the natural habitat. Incubation under in situ pressure (200 atm [1 atm = 101.29 kPa]) did not increase the maximum growth temperature by more than 1 degrees C for any of the organisms but did result in increases in growth rates of up to 15% at optimum growth temperatures. At in situ pressure, temperatures considerably higher than those limiting growth (i.e., > 105 degrees C) were survived best by isolates with the highest maximum growth temperatures, but none of the organisms survived at temperatures of 150 degrees C or higher for 5 min. Free oxygen was toxic to all isolates at growth range temperatures, but at ambient deep-sea temperature (3 to 4 degrees C), the effect varied in different isolates, the non-sulfur-dependent isolate being the most oxygen tolerant. Hyperthermophiles could be isolated from refrigerated and oxygenated samples after 5 years of storage. Cu, Zn, and Pb ions were found to be toxic under nongrowth conditions (absence of organic substrate), with the non-sulfur-dependent isolate again being the most tolerant. PMID- 16348800 TI - Heteropolysaccharide Formation by Arthrobacter viscosus Grown on Xylose and Xylose Oligosaccharides. AB - Arthrobacter viscosus NRRL B-1973 produces its viscous extracellular polysaccharides (EPS) when grown on media containing xylose or enzymatic xylan hydrolysates. Crude EPS formation from xylose averaged 12 g/liter when initial culture pH was adjusted to 8.0 and total nitrogen was limited to 0.03%. Purified EPS from pentose and hexose substrates were analyzed for their monosaccharide, acetyl, and uronic acid components, intrinsic viscosities, and average molecular masses. Differences were apparent in degrees of acetylation, molecular masses, and intrinsic viscosities of the heteropolysaccharides produced on different carbon sources. PMID- 16348801 TI - Identification of distinct communities of sulfate-reducing bacteria in oil fields by reverse sample genome probing. AB - Thirty-five different standards of sulfate-reducing bacteria, identified by reverse sample genome probing and defined as bacteria with genomes showing little or no cross-hybridization, were in part characterized by Southern blotting, using 16S rRNA and hydrogenase gene probes. Samples from 56 sites in seven different western Canadian oil field locations were collected and enriched for sulfate reducing bacteria by using different liquid media containing one of the following carbon sources: lactate, ethanol, benzoate, decanoate, propionate, or acetate. DNA was isolated from the enrichments and probed by reverse sample genome probing using master filters containing denatured chromosomal DNAs from the 35 sulfate reducing bacterial standards. Statistical analysis of the microbial compositions at 44 of the 56 sites indicated the presence of two distinct communities of sulfate-reducing bacteria. The discriminating factor between the two communities was the salt concentration of the production waters, which were either fresh water or saline. Of 34 standards detected, 10 were unique to the fresh water and 18 were unique to the saline oil field environment, while only 6 organisms were cultured from both communities. PMID- 16348802 TI - Investigation of Mitochondrial Transmission in Selected Matings between Homokaryons from Commercial and Wild-Collected Isolates of Agaricus bisporus (= Agaricus brunnescens). AB - Ten heterokaryons of Agaricus bisporus (= Agaricus brunnescens) were shown to carry four different mitochondrial (mt) genotypes by analysis of mt restriction fragment length polymorphisms (RFLPs). Fifteen homokaryons derived from these strains were used to investigate mt inheritance in A. bisporus. One hundred eighty-nine pairings were performed in 25 different combinations. Pairings in 15 different combinations produced heterokaryons on the basis of nuclear RFLP analyses and/or fruiting trials. The mt genotype of each new intraspecies hybrid was examined by using mt RFLPs as genetic markers. Our results suggest the following. (i) Recombination between the mt genomes was not a common event. (ii) From most individual pairings, all heterokaryons carried the same mt genotype. (iii) Heterokaryons carrying either of the two possible mt genotypes were observed in certain crosses after modification of the pairing procedure. A biparental transmission pattern was demonstrated for some crosses, but there appears to be a preference for one of the mt genotypes to predominate in any specific pairing. PMID- 16348803 TI - Methods involving light variation for isolation of cyanobacteria: characterization of isolates from central australia. AB - We report the isolation of organisms belonging to a range of cyanobacterial genera from the remote arid region of central Australia, together with a preliminary characterization of their temporal modes of nitrogen fixation. We rendered unialgal Dermocarpa and Myxosarcina spp. (Section II organisms), LPP group B: type X (Section III), and Scytonema and Nostoc spp. (Section IV). We developed an isolation procedure based on a combination of published methods and applicable to a broad spectrum of cyanobacterial genera. We found light intensity to be a critical variable in our methodology; it was also an important determinant of the proportions of different organisms growing in stable mixtures. PMID- 16348804 TI - Synthesis of Poly(3-Hydroxybutyrate-Co-3-Hydroxyvalerate) from Methanol and n Amyl Alcohol by the Methylotrophic Bacteria Paracoccus denitrificans and Methylobacterium extorquens. AB - Strains of two types of methylotrophic bacteria, Paracoccus denitrificans and Methylobacterium extorquens, synthesized the copolyester poly(3-hydroxybutyrate co-3-hydroxyvalerate) when methanol and n-amyl alcohol were added together to nitrogen-limited medium. The composition of the copolyester differed considerably between the two strains: the copolyester from P. denitrificans was comparatively rich in 3-hydroxyvalerate (3HV). The 3HV content of the copolyester synthesized by this strain increased with increasing concentrations of n-amyl alcohol. Its maximum content was 91.5 mol% under the conditions used. In M. extorquens, the maximum 3HV content was limited to 38.2 mol%. Since n-amyl alcohol served as a substrate for a standard methanol dehydrogenase, the enzyme was proposed to oxidize both methanol and n-amyl alcohol in the first step of copolyester synthesis from these substrates by methanol-grown cells. PMID- 16348805 TI - Purification of Two Nitrate Reductases from Xanthomonas maltophilia Grown in Aerobic Cultures. AB - Xanthomonas maltophilia ATCC 17666 is an obligate aerobe that accumulates nitrite when grown on nitrate. Spectra of membranes from nitrate-grown cells exhibited b type cytochrome peaks and A(615-630) indicative of d-type cytochrome but no absorption peaks corresponding to c-type cytochromes. The nitrate reductase (NR) activity was located in the membrane fraction. Triton X-100-extracted reduced methyl viologen-NRs were purified on DE-52, hydroxylapatite, and Sephacryl S-300 columns to specific activities of 52 to 67 mumol of nitrite formed per min per mg of protein. The cytochrome-containing NR(I) separated on sodium dodecyl sulfate polyacrylamide gel electrophoresis into a 135-kDa alpha-subunit, a 64-kDa beta subunit, and a 23-kDa gamma-subunit with relative band intensities indicative of a 1:1:1 alpha/beta/gamma subunit ratio and a M(r) of 222,000. The electronic spectrum of dithionite-reduced purified NR displayed peaks at 425, 528, and 558 nm, indicative of the presence of a cytochrome b, an interpretation consistent with the pyridine hemochrome spectrum formed. The cytochrome b of the NR was reduced under anaerobic conditions by menadiol and oxidized by nitrate with the production of nitrite. This NR contained 0.96 Mo, 12.5 nonheme iron, and 1 heme per 222 kDa: molybdopterin was detected with the Neurospora crassa nit-1 assay. A smaller reduced methyl viologen-NR (169 kDa), present in various concentrations in the Triton X-100 preparations, lacked a cytochrome spectrum and did not oxidize menadiol. The characteristics of the NRs and the absence of c-type cytochromes provide insights into why X. maltophilia accumulates nitrite. PMID- 16348806 TI - Assessment of [h]thymidine incorporation into DNA as a method to determine bacterial productivity in stream bed sediments. AB - We performed several checks on the underlying assumptions and procedures of the thymidine technique applied to stream bed sediments. Bacterial production rates were not altered when sediments were mixed to form a slurry. Incubation temperature did affect production rates. Controls fixed and washed with formaldehyde had lower backgrounds than trichloroacetic acid controls. DNA extraction by base hydrolysis was incomplete and variable at 25 degrees C, but hydrolysis at 120 degrees C extracted 100% of the DNA, of which 84% was recovered upon precipitation. Production rates increased as thymidine concentrations were increased over 3 orders of magnitude (30 nM to 53 muM thymidine). However, over narrower concentration ranges, thymidine incorporation into DNA was independent of thymidine concentration. Elevated exogenous thymidine concentrations did not eliminate de novo synthesis. Transport of thymidine into bacterial cells occurred at least 5 to 20 times faster than incorporation of label into DNA. We found good agreement between production rates of bacterial cultures based upon increases in cell numbers and estimates based upon thymidine incorporation and amount of DNA per cell. Those comparisons emphasized the importance of isotopic dilution measurements and validated the use of the reciprocal plot technique for estimating isotopic dilution. Nevertheless, the thymidine technique cannot be considered a routine assay and the inability to measure the cellular DNA content in benthic communities restricts the accuracy of the method in those habitats. PMID- 16348807 TI - Incorporation of [h]leucine and [h]valine into protein of freshwater bacteria: uptake kinetics and intracellular isotope dilution. AB - Incorporation of [H]leucine and [H]valine into proteins of freshwater bacteria was studied in two eutrophic lakes. Incorporation of both amino acids had a saturation level of about 50 nM external concentration. Only a fraction of the two amino acids taken up was used in protein synthesis. At 100 nM, the bacteria respired 91 and 78% of leucine and valine taken up, respectively. Respiration of H and C isotopes of leucine gave similar results. Most of the nonrespired leucine was recovered in bacterial proteins, while only up to one-half of the nonrespired valine occurred in proteins. In intracellular pools of the bacteria, [H]leucine reached an isotope saturation of 88 to 100% at concentrations of >40 nM. For [H]valine, an isotope equilibrium of about 90% was obtained at concentrations of >80 nM. Within an incubation period of typically 1 h, tritiated leucine and valine incorporated into proteins of the bacteria reached an isotope saturation of 2 to 6%. In a 99-h batch experiment, bacterial protein synthesis calculated from incorporation of leucine and valine corresponded to 31 and 51% (10 nM) and 89 and 97% (100 nM), respectively, of the chemically determined protein production. Measured conversion factors of 100 nM leucine and valine were 6.4 x 10 and 6.6 x 10 cells per mol, respectively, and fell within the expected theoretical values. The present study demonstrates that incorporation of both valine and leucine produces realistic measurements of protein synthesis in freshwater bacteria and that the incorporation can be used as a measure of bacterial production. PMID- 16348808 TI - Incorporation of [h]leucine and [h]valine into protein of freshwater bacteria: field applications. AB - Incorporation of leucine and valine into proteins of freshwater bacteria as a measure of bacterial production was tested in two eutrophic Danish lakes and was related to bacterial production measured by thymidine incorporation. In a depth profile (0 to 8 m) in Frederiksborg Castle Lake, incorporation of 100 nM leucine and valine gave similar rates of protein production. In terms of carbon, this production was about 50% lower than incorporation of 10 nM thymidine. In another depth profile in the same lake, incorporations of 10 nM valine and 100 nM leucine were identical, but differed from incorporations of 10 nM leucine and 100 nM valine. Bacterial carbon production calculated from incorporations of 10 nM thymidine and 10 nM leucine was similar, whereas 10 nM valine and 100 nM leucine and valine indicated an up to 2.4-fold-higher rate of carbon production. In a diel study in Lake Bagsvaerd, incorporation of 100 nM leucine and valine indicated a similar protein production, but the calculated carbon production was about 1.9-fold higher than the production based on uptake of 10 nM thymidine. Different diel changes in incorporation of the two amino acids and in incorporation of thymidine were observed. In both lakes, concentrations of naturally occurring leucine and valine were <5 nM in most samples. This means that the specific activity of a H isotope added at a concentration of 100 nM usually was diluted a maximum of 5%. Net assimilation of natural free amino acids in the lakes sustained 8 to 69% of the net bacterial carbon requirement, estimated from incorporation of leucine, valine, or thymidine. The present results indicate that incorporation of leucine and valine permits realistic measurements of bacterial production in freshwater environments. PMID- 16348809 TI - Ultrastructural and Immunocytochemical Studies on the H(2)O(2)-Producing Enzyme Pyranose Oxidase in Phanerochaete chrysosporium Grown under Liquid Culture Conditions. AB - The ultrastructural distribution of the sugar-oxidizing enzyme pyranose 2-oxidase (POD) in hyphae of Phanerochaete chrysosporium K-3 grown under liquid culture conditions optimal for the enzyme's production was studied by transmission electron microscopy immunocytochemistry. Using the 3-dimethylaminobenzoic acid-3 methyl-2-benzothiazolinone hydrazone hydrochloride H(2)O(2) peroxidase spectrophotometric assay, POD was detected in mycelial extracts from days 7 to 18, with maximum activity recorded on day 12. Onset of POD activity occurred in the secondary phase of hyphal development at a time of stationary growth, glucose limitation, and pH increase. POD was also detected extracellularly in the culture fluid from days 7 to 18, with maximum activity recorded on day 13. At early stages of development (3 to 4 days), using anti-POD antibodies and immunogold labeling, POD was localized in multivesicular and electron-dense bodies and in cell membrane regions. After 10 to 12 days of growth, at maximum POD activity, POD was concentrated within the periplasmic space where it was associated with membrane-bound vesicles and other membrane structures. At later stages of development (17 to 18 days), when the majority of hyphae were lysed, POD was observed associated with residual intracellular membrane systems and vesicles. Transmission electron microscopy immunocytochemical studies also demonstrated an extracellular distribution of the enzyme at the stationary growth phase, showing its association with fungal extracellular slime. In studies of ligninolytic cultures of the same fungus, POD was found to have a similar intracellular and extracellular distribution in slime as that recorded for cultures grown with cornsteep. POD's peripheral cytoplasmic distribution shows similarities to the cellular distribution of that reported previously for H(2)O(2)-dependent lignin and manganese peroxidases in P. chrysosporium. PMID- 16348810 TI - Methylosinus trichosporium OB3b Mutants Having Constitutive Expression of Soluble Methane Monooxygenase in the Presence of High Levels of Copper. AB - The methanotrophic bacterium Methylosinus trichosporium OB3b is unusually active in degrading recalcitrant haloalkanes such as trichloroethylene (TCE). The first and rate-limiting step in the degradation of TCE is catalyzed by a soluble methane monooxygenase (sMMO). This enzyme is not expressed when the cells are grown in the presence of copper at concentrations typically found in polluted groundwater. Under these conditions, M. trichosporium OB3b expresses a particulate form of the enzyme (pMMO), which has a narrow substrate specificity and does not degrade TCE at any significant rate. We have isolated M. trichosporium OB3b mutants that are deficient in pMMO and express sMMO constitutively in the presence of elevated concentrations of copper. One mutant (PP358) exhibited a TCE degradation rate which was almost twice as high as that of the wild-type strain grown under optimal conditions (without copper). All of the mutants lost the ability to express pMMO activity and to form stacked intracellular membranes characteristic of wild-type cells expressing pMMO. PMID- 16348811 TI - Direct and indirect evidence of size-selective grazing on pelagic bacteria by freshwater nanoflagellates. AB - Size-selective grazing of three heterotrophic nanoflagellates (with cell sizes of 21, 44, and 66 mum) isolated from Lake Arlington, Texas was examined by using a natural mixture of fluorescence labelled lake bacteria. Sizes of ingested bacteria in food vacuoles were directly measured. Larger bacterial cells were ingested at a frequency much higher than that at which they occurred in the assemblage, indicating preferential flagellate grazing on the larger size classes within the lake bacterioplankton. Water samples were collected biweekly from June through September, 1989, fractionated by filtration, and incubated for 40 h at in situ temperatures. The average bacterial size was always larger in water which was passed through 1-mum-pore-size filters (1-mum-filtered water) (which was predator free) than in 5-mum-filtered water (which contained flagellates only) or in unfiltered water (in which all bacterivores were present). The increase of bacterial-cell size in 1-mum-filtered water was caused by a shift in the size structure of the bacterioplankton population. Larger cells became more abundant in the absence of flagellate grazing. PMID- 16348812 TI - Mechanisms and rates of decay of marine viruses in seawater. AB - Loss rates and loss processes for viruses in coastal seawater from the Gulf of Mexico were estimated with three different marine bacteriophages. Decay rates in the absence of sunlight ranged from 0.009 to 0.028 h, with different viruses decaying at different rates. In part, decay was attributed to adsorption by heat labile particles, since viruses did not decay or decayed very slowly in seawater filtered through a 0.2-mum-pore-size filter (0.2-mum-filtered seawater) and in autoclaved or ultracentrifuged seawater but continued to decay in cyanide-treated seawater. Cyanide did cause decay rates to decrease, however, indicating that biological processes were also involved. The observations that decay rates were often greatly reduced in 0.8- or 1.0-mum-filtered seawater, whereas bacterial numbers were not, suggested that most bacteria were not responsible for the decay. Decay rates were also reduced in 3-mum-filtered or cycloheximide-treated seawater but not in 8-mum-filtered seawater, implying that flagellates consumed viruses. Viruses added to flagellate cultures decayed at 0.15 h, corresponding to 3.3 viruses ingested flagellate h. Infectivity was very sensitive to solar radiation and, in full sunlight, decay rates were 0.4 to 0.8 h. Even when UV-B radiation was blocked, rates were as high as 0.17 h. Calculations suggest that in clear oceanic waters exposed to full sunlight, most of the virus decay, averaged over a depth of 200 m, would be attributable to solar radiation. When decay rates were averaged over 24 h for a 10-m coastal water column, loss rates of infectivity attributable to sunlight were similar to those resulting from all other processes combined. Consequently, there should be a strong diel signal in the concentration of infectious viruses. In addition, since sunlight destroys infectivity more quickly than virus particles, a large proportion of the viruses in seawater is probably not infective. PMID- 16348813 TI - Release of bacterial DNA by marine nanoflagellates, an intermediate step in phosphorus regeneration. AB - The concentrations of dissolved DNA and nanoflagellates were found to covary during a study of diel dynamics of the microbial food web in the Adriatic Sea. This observation was further investigated in a continuous seawater culture when nanoflagellates were fed bacteria grown in filtered seawater. Analysis of dissolved organic phosphorus and dissolved DNA showed a sixfold increase of dissolved DNA in the presence of the nanoflagellates (Ochromonas sp.). The amount of DNA released suggested that the majority of the consumed bacterial DNA was ejected. Phagotrophic nanoflagellates thus represent an important source of origin for dissolved DNA. The rate of breakdown of dissolved DNA and release of inorganic phosphorus in the pelagic ecosystem is suggested to be dependent on the ambient phosphate pool. In the P-limited northern Adriatic Sea, rapid degradation of the labelled DNA could be demonstrated, whereas the N-limited southern California bight water showed a much lower rate. Phosphorus originating from dissolved DNA was shown to be transferred mainly to organisms in the <3-mum-size fractions. On the basis of the C/P ratios, we suggest that a significant fraction of the phosphorus demand by the autotrophs may be sustained by the released DNA during stratified conditions. Thus, the nucleic acid-rich bacterial biomass grazed by protozoa plays an important role in the biogeochemical cycling of phosphorus in the marine environment. PMID- 16348814 TI - Oxidation of Elemental Sulfur to Sulfite by Thiobacillus thiooxidans Cells. AB - Thiobacillus thiooxidans cells oxidized elemental sulfur to sulfite, with 1 mol of O(2) consumption per mol of sulfur oxidized to sulfite, when the oxidation of sulfite was inhibited with 2-n-heptyl-4-hydroxyquinoline N-oxide. PMID- 16348815 TI - Production of Eicosapentaenoic Acid (20:5 n-3) by Vibrio pelagius Isolated from Turbot (Scophthalmus maximus (L.)) Larvae. AB - Fourteen bacterial strains isolated from turbot, Scophthalmus maximus (L.), larvae were screened for eicosapentaenoic acid (20:5 n-3) (EPA) production. Gas chromatography analysis revealed that one bacterial species, Vibrio pelagius, contained a high proportion of EPA in cellular lipid. This finding was confirmed by gas chromatography-mass spectrometry analysis. A higher concentration of EPA was detected when the bacterium was cultured at 4 degrees C. PMID- 16348816 TI - Evidence for methylotrophic symbionts in a hydrothermal vent mussel (bivalvia: mytilidae) from the mid-atlantic ridge. AB - Symbioses between chemolithoautotrophic bacteria and the major macrofaunal species found at hydrothermal vents have been reported for numerous sites in the Pacific Ocean. We present microscopical and enzymatic evidence that methylotrophic bacteria occur as intracellular symbionts in a new species of mytilid mussel discovered at the Mid-Atlantic Ridge hydrothermal vents. Two distinct ultrastructural types of gram-negative procaryotic symbionts were observed within gill epithelial cells by transmission electron microscopy: small coccoid or rod-shaped cells and larger coccoid cells with stacked intracytoplasmic membranes typical of methane-utilizing bacteria. Methanol dehydrogenase, an enzyme diagnostic of methylotrophs, was detected in the mytilid gills, while tests for ribulose-1,5-bisphosphate carboxylase, the enzyme diagnostic of autotrophy via the Calvin cycle, were negative. Stable carbon isotope values (deltaC) of mytilid tissue (-32.7 and -32.5% for gill and foot tissues, respectively) fall within the range of values reported for Pacific vent symbioses but do not preclude the use of vent-derived methane reported to be isotopically heavy relative to biogenically produced methane. PMID- 16348817 TI - Effect of Light and Organic Acids on Oxygen Uptake by BTAi 1, a Photosynthetic Rhizobium. AB - A photosynthetic rhizobium, strain BTAi 1, was cultured ex planta to investigate its photosynthetic-respiratory system and the response of this interactive system to light quantity and quality and to the addition of organic acids. Oxygen uptake, as measured with an oxygen electrode, is diminished upon illumination, with the amount of decrease related to light intensity. This oxygen-sparing effect is correlated with the wavelengths of light that are associated with bacteriochlorophyll absorbance. Increasing concentrations of glutamic, succinic, and malic acids enhance the oxygen-sparing effect of light until a threshold concentration is reached, beyond which succinic and malic acids decrease the effect. The photobiology of this unique rhizobium is similar to the photobiology of both anaerobic and aerobic photosynthetic bacteria. PMID- 16348818 TI - Competitive Growth of Genetically Marked Malolactic-Deficient Lactobacillus plantarum in Cucumber Fermentations. AB - Procedures were developed for the differential enumeration of an added strain of Lactobacillus plantarum and indigenous lactic acid bacteria (LAB) during the fermentation of brined cucumbers. The added strain was an N,N-nitrosoguanidine generated mutant that lacked the ability to produce CO(2) from malic acid (MDC). The MDC phenotype is desirable because CO(2) production from malic acid decarboxylation has been shown to contribute to bloater formation in fermented cucumbers. A basal medium containing malic acid and adjusted to pH 4.0 permitted growth of indigenous LAB (predominantly MDC), but not growth of the added MDC culture. Transformation of the MDC culture by electroporation with cloning vector pGK12 conferred chloramphenicol resistance, which permitted selective enumeration of this culture. The reversion frequency of the MDC mutation was determined by a fluctuation test to be less than 10. The level of retention of plasmid pGK12 was greater than 90% after 10 generations in cucumber juice medium at 32 degrees C. With the procedures developed, we were able to establish the ratio of MDC to MDC LAB that results in malic acid retention in fermentations of filter-sterilized cucumber juice and unsterilized whole cucumbers under specified conditions. PMID- 16348819 TI - Comparison of rates of flagellate bacterivory and bacterial production in a marine coastal system. AB - Protozoan predation on bacteria and bacterioplankton secondary production were simultaneously determined in La Salvaje Beach water during 1990. Protozoan grazing on bacterioplankton was measured from fluorescently labeled bacterium uptake rates; estimates of bacterial secondary production were obtained from [H]thymidine incorporation rates. Two different conversion factors were used to transform thymidine incorporation rates into bacterial production rates; both of them were specific for La Salvaje Beach and were calculated by using empirical and semitheoretical approaches. The average flagellate predation rate was 14.0 bacteria flagellate h; the average population predation rate was 7.35 x 10 bacteria liter h. The estimates of bacterial production differed greatly depending on the conversion factor used, and so did the percentages of bacterial production consumed by flagellated protozoa (4.6% when the empirical conversion factor for La Salvaje Beach was used and 113% when the semitheoretical conversion factor specific for this system was used). The ecological implications of each of these values are discussed. PMID- 16348820 TI - Competition between Two Isolates of Denitrifying Bacteria Added to Soil. AB - We examined the competitive relationship between two isolates of denitrifying bacteria, both of which grow well under aerobic conditions but differ in their ability to grow under denitrifying conditions. The growth and persistence of the two isolates, added to sterile soil or added to soil previously colonized by the other isolate, were monitored under aerobic and denitrifying (anaerobic) conditions. When isolates were added together to sterile soil, the isolate added at the higher density reduced the growth of the isolate added at the lower density. The magnitude of the growth reduction varied depending on the competitive abilities of the individual isolates and the aeration state of the soil. Prior colonization of soil with one of the isolates conferred a competitive advantage on the colonized isolate but did not lead to the disappearance of the challenging isolate. Fluctuations in aeration state caused large changes in the population density of one isolate and altered the competitive relationship between the two isolates. The competitive effectiveness of each isolate varied with cell density, the degree of prior colonization of the soil by the other isolate, and the aeration state of the soil. PMID- 16348821 TI - Physiological Events in Clostridium acetobutylicum during the Shift from Acidogenesis to Solventogenesis in Continuous Culture and Presentation of a Model for Shift Induction. AB - The pH of continuous cultures of Clostridium acetobutylicum growing at pH 5.6 was allowed to decrease to 4.3 after acid production and thereby to shift the cultures from acetate and butyrate to acetone and butanol formation. Several parameters were determined during the shift. An increase in the intracellular acid concentration to 440 mM was recorded. An excess of undissociated butyric acid but not of acetic acid just before the shift to solventogenesis was followed by a decline in acid production and subsequently by the uptake of acids. The intracellular ATP concentration reached a minimum before the onset of solventogenesis; this presumably reflects the ATP-consuming proton extrusion connected with the increase in the DeltapH from 0.7 to 1.4 units. The pool of NADH plus NADPH exhibited a drastic increase until solventogenesis was induced. The changes in the ATP and ADP and NADH plus NADPH pools during these pH shift experiments were the beginning of a stable metabolic oscillation which could also be recorded as an oscillation of the culture redox potential under steady-state solventogenic conditions. Similar changes were observed when the shift was induced by the addition of butyrate and acetate (50 mM each) to the continuous culture. However, when methyl viologen was added, important differences were found: ATP levels did not reach a minimum, acetoacetate decarboxylase activity could not be measured, and butanol but not acetone was produced. A model for the shift is proposed; it assumes the generation of two signals, one by the changed ATP and ADP levels and the other by the increased NAD(P)H level. PMID- 16348822 TI - Production of the Mycotoxin Fumonisin B(1) by Alternaria alternata f. sp. lycopersici. AB - The mycotoxin fumonisin B(1), originally described as being produced by Fusarium moniliforme, was detected in liquid cultures of Alternaria alternata f. sp. lycopersici, a host-specific pathogen of tomato plants. The metabolite was detected by high-pressure liquid chromatography and mass spectrometry. Its identity was confirmed by fast atom bombardment and ion spray mass spectrometry, as well as parent-daughter tandem mass spectrometry. In three separate experiments, the concentrations found ranged between 5 and 140 ppm (mug/ml). PMID- 16348823 TI - Diversity among Streptomyces Strains Causing Potato Scab. AB - Eighty Streptomyces isolates, including 35 potato scab-inducing strains and 12 reference strains of Streptomyces scabies, were physiologically characterized by a total of 329 miniaturized tests. Overall similarities of all strains were determined by numerical taxonomy, with the unweighted average linkage (UPGMA) algorithm and simple matching (S(sm)) and Jaccard (S(j)) coefficients used as measures for similarity. Three cluster groups (A to C) were defined at a similarity level of 80.1% (S(sm)); these groups contained 14 clusters and 24 unclustered strains defined at a similarity level of 86.5% (S(sm)). Cluster group A contained strains phenotypically related to S. griseus or S. exfoliatus, whereas cluster group B contained strains which were phenotypically related to S. violaceus or S. rochei. The majority of the pathogenic isolates and reference strains were assigned to S. violaceus (57%) and S. griseus (22%). A DNA probe derived from the rRNA operon of S. coelicolor IMET 40271 was used to detect restriction fragment length polymorphisms (RELPs) among 40 pathogenic and nonpathogenic Streptomyces isolates. Southern blots revealed a high degree of diversity among the pathogenic strains tested. No significant correlation between numerical classification and RFLP grouping of Streptomyces strains could be revealed. The results obtained suggest that RFLP data are of minor importance in classification of Streptomyces species and that genes for pathogenicity determinants are spread among different Streptomyces species by mobilizable elements. PMID- 16348824 TI - Improved Methods for Cultivation of the Extremely Thermophilic Bacterium Thermotoga neapolitana. AB - Growth medium components and cultivation conditions for the extremely thermophilic bacterium Thermotoga neapolitana were optimized. A defined marine salts medium was formulated. Trace amounts of iron stimulated growth of T. neapolitana, while zinc inhibited growth at concentrations exceeding 11.1 muM. Other trace metals had no effect on its growth. Of the vitamins tested, only biotin was required for optimal growth. A defined mineral medium containing 5 g of carbohydrates per liter as the carbon source and 0.5 g of cysteine per liter as the sulfur source and reductant supported growth. Growth was stimulated by inclusion of vitamin-free Casamino Acids. Elemental sulfur, cystine, and dimethyl disulfide in the growth medium enhanced growth. Elemental sulfur and cystine relieved growth inhibition by hydrogen. T. neapolitana formed colonies in 2 days on plates of complex medium solidified with gellan gum and in 4 days on defined medium. The efficiency of plating was determined when growing cultures were sampled both aerobically and anaerobically and plated under aerobic and anaerobic conditions. Mean plating efficiencies were improved by sampling the growing cultures under strictly anaerobic conditions. Little or no improvement was obtained by inoculating plates inside an anaerobic chamber. Plating efficiencies of approximately 80% were obtained. Polycarbonate jars with aluminum lids withstood repeated incubation at 77 degrees C without significant deterioration of the anaerobic seal and provided the most consistent results. PMID- 16348825 TI - Production of pyrroloquinoline quinone by using methanol-utilizing bacteria. AB - A large number of methanol-utilizing bacteria were screened for extracellular production of pyrroloquinoline quinone (PQQ) by using methanol as the carbon and energy sources. Of the bacteria selected, Hyphomicrobium sp. strain TK 0441 was examined for PQQ production by using a jar fermentor. The amount of PQQ in the broth and the level of methanol dehydrogenase activity in the cells were increased by simply decreasing the amount of Fe added to the medium. On the other hand, extracellularly produced protein which interfered with the purification of PQQ was decreased by simply increasing the amount of Mg added to the medium. A suitable medium that contained 1 mug of Fe per ml, 150 mug of Mg per ml, and trace elements was developed. In this medium, the production of PQQ reached approximately 1 mg/ml and protein formation was low. PMID- 16348826 TI - Isolation, Cultural Maintenance, and Taxonomy of a Sheath-Forming Strain of Leptothrix discophora and Characterization of Manganese-Oxidizing Activity Associated with the Sheath. AB - Leptothrix discophora SP-6 was isolated from the outflow reservoir of an artificial iron seep. Its sheathforming phenotype was maintained by slow growth in a mineral salts-vitamin-pyruvate medium under minimal aeration at 20 to 25 degrees C. A sheathless variant, SP-6(sl), was isolated from smooth colonies that appeared on spread plates after rapid growth of SP-6 in well-aerated cultures. SP 6 and SP-6(sl) are closely related but not identical to the previously studied sheathless strain SS-1 (ATCC 43182). Increasing Mn concentrations in the growth medium of SP-6 increased the phase density of the sheath, indicating increased Mn oxide deposition in the sheath. Electron microscopy of cultures grown without added Mn revealed that the sheath consisted of a well-defined inner layer, 30 to 100 nm thick, and a diffuse outer capsular layer of variable thickness. Mn oxides were identified in the sheath by their characteristic ultrastructure, electron density, and X-ray-dispersive energy spectra. In heavily encrusted sheaths, the Mn oxides were evenly distributed in both layers of the sheath. Sheathed cells retained more Mn-oxidizing activity than did sheathless cells after washing with distilled, deionized water; the sheath retained some of its activity after an EDTA-lysozyme-detergent treatment which removed the cells. An ultrafiltration dialysis procedure significantly increased the recovery of activity from spent media of SP-6 over that reported previously for SS-1 (L.F. Adams and W.C. Ghiorse, J. Bacteriol. 169:1279-1285, 1987). A 108-kDa Mn-oxidizing protein was identified in concentrated spent media of SP-6 and SP-6(sl), and the activity of the concentrates showed stability in detergents comparable to that of SS-1 and patterns of heat inactivation and chemical inhibition similar to those of SS-1. PMID- 16348827 TI - Evaluation of Culture Techniques for Isolation of Pseudomonas pseudomallei from Soil. AB - Three selective enrichment broths and four selective agar media were evaluated for their ability to support the growth of Pseudomonas pseudomallei both at 35 degrees C and at ambient temperature (range, 20 to 32 degrees C; mean, 25 degrees C). Colony counts of 50 strains of P. pseudomallei and recovery studies with 1 soil strain in 60 simulated soil samples demonstrated that enrichment with Trypticase soy broth incorporating 5 mg of crystal violet per liter and 20 mg of colistin per liter (CVCB) and subculture to Ashdown medium supported the growth of all 50 strains and produced the highest recovery rates with the greatest suppression of other soil flora. An enrichment broth of MacConkey broth (purple) incorporating 10 mg of crystal violet per liter, 5 mg of bromcresol purple per liter, 25 mg of gentamicin per liter, and 650 mg of streptomycin per liter showed greater suppression of soil bacteria than CVCB, but it failed to support the growth of three strains of P. pseudomallei. Recovery rates were essentially the same irrespective of whether the soil samples were incubated at 35 degrees C or at ambient temperature, provided cultures were incubated in protected shade for an extended period. This is an important feature for field work in large-scale epidemiological surveys in which resources are limited. PMID- 16348828 TI - Soil Bacterial Biomass, Activity, Phospholipid Fatty Acid Pattern, and pH Tolerance in an Area Polluted with Alkaline Dust Deposition. AB - Soil bacterial biomass, phospholipid fatty acid pattern, pH tolerance, and growth rate were studied in a forest area in Finland that is polluted with alkaline dust from an iron and steel works. The pollution raised the pH of the humus layer from 4.1 to 6.6. Total bacterial numbers and the total amounts of bacterial phospholipid fatty acids in the humus layer did not differ between the unpolluted control sites and the polluted ones. The number of CFU increased by a factor of 6.4 in the polluted sites compared with the controls, while the bacterial growth rate, measured by the thymidine incorporation technique, increased about 1.8-fold in the polluted sites. A shift in the pattern of phospholipid fatty acids indicated a shift in the bacterial species composition. The largest proportional increase was found for the fatty acid 10Me18:0, which indicated an increase in the number of actinomycetes in the polluted sites. The levels of the fatty acids i14:0, 16:1omega5, cy17:0, 18:1omega7, and 19:1 also increased in the polluted sites while those of fatty acids 15:0, i15:0, 10Me16:0, 16:1omega7t, 18:1omega9, and cy19:0 decreased compared with the unpolluted sites. An altered pH tolerance of the bacterial assemblage was detected either as a decrease in acid-tolerant CFU in the polluted sites or as altered bacterial growth rates at different pHs. The latter was estimated by measuring the thymidine incorporation rate of bacteria extracted from soil by homogenization-centrifugation at different pHs. PMID- 16348829 TI - Reduction of Selenate and Selenite to Elemental Selenium by a Pseudomonas stutzeri Isolate. AB - A Pseudomonas stutzeri isolate rapidly reduced both selenite and selenate ions to elemental selenium at initial concentrations of both anions of up to 48.1 mM. Optimal selenium reduction occurred under aerobic conditions between pH 7.0 and 9.0 and at temperatures of 25 to 35 degrees C. Reduction of both selenite and selenate was unaffected by a number of anions except for sulfite, chromate, and tungstate ions, which inhibited both growth and reduction. PMID- 16348830 TI - Divergence of Genomic Sequences between Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris. AB - Relatedness between Lactococcus lactis subsp. cremoris and L. lactis subsp. lactis was assessed by Southern hybridization analysis, with cloned chromosomal genes as probes. The results indicate that strains of the two subspecies form two distinct groups and that the DNA sequence divergence between L. lactis subsp. lactis and L. lactis subsp. cremoris is estimated to be between 20 and 30%. The previously used phenotypic criteria do not fully discriminate between the groups; therefore, we propose a new classification which is based on DNA homology. In agreement with this revised classification, the L. lactis subsp. lactis and L. lactis subsp. cremoris strains from our collection have distinct phage sensitivities. PMID- 16348831 TI - Toxicity of pentachlorophenol to six species of white rot fungi as a function of chemical dose. AB - The growth of six species of white rot fungi was a function of pentachlorophenol (PCP) dose, expressed as mass of PCP per mass of mycelia, at PCP doses SO(4) + 3H(2)S + 2H. Subsequent chemical reactions between the formed sulfide and the added FeOOH led to the observed precipitation of iron sulfides. Sulfate and iron sulfides were also produced when FeOOH was replaced by FeCO(3). Further enrichment with manganese oxide, MnO(2), instead of FeOOH yielded stable cultures which formed sulfate during concomitant reduction of MnO(2) to Mn. Growth of small rod-shaped bacteria was observed. When incubated without MnO(2), the culture did not grow but produced small amounts of SO(4) and H(2)S at a ratio of 1:3, indicating again a disproportionation of S. The observed microbial disproportionation of S only proceeds significantly in the presence of sulfide scavenging agents such as iron and manganese compounds. The population density of bacteria capable of S disproportionation in the presence of FeOOH or MnO(2) was high, > 10 cm in coastal sediments. The metabolism offers an explanation for recent observations of anaerobic sulfide oxidation to sulfate in anoxic sediments. PMID- 16348836 TI - Influence of complex structure on the biodegradation of iron-citrate complexes. AB - The biodegradation of iron-citrate complexes depends on the structure of the complex formed between the metal and citric acid. Ferric iron formed a bidentate complex with citric acid, [Fe(III) (OH)(2) cit] involving two carboxylic acid groups, and was degraded at the rate of 86 muM h. In contrast, ferrous iron formed a tridentate complex with citric acid, [Fe(II) cit], involving two carboxylic acid groups and the hydroxyl group, and was resistant to biodegradation. However, oxidation and hydrolysis of the ferrous iron resulted in the formation of a tridentate ferric-citrate complex, [Fe(III)OH cit], which was further hydrolyzed to a bidentate complex, [Fe(III)(OH)(2) cit], that was readily degraded. The rate of degradation of the ferrous-citrate complex depended on the rate of its conversion to the more hydrolyzed form of the ferric-citrate complex. Bacteria accelerated the conversion much more than did chemical oxidation and hydrolysis. PMID- 16348837 TI - Glucose uptake and end product formation in an intertidal marine sediment. AB - Glucose uptake was monitored on a seasonal basis, using [6-H]glucose and undisturbed cores collected from an intertidal mud flat. The fate of glucose carbon, including the formation of CO(2) and biomass, was assayed by using undisturbed cores and [U-C]glucose; the production of short-chain fatty acids was monitored with [U-C]glucose and sediment slurries. Rate constants for glucose uptake varied temporally, with temperature accounting for much of the variability; turnover times ranged from about 2 to 10 min. Rate constants decreased with increasing sediment depth and in the following order for several common monosaccharides: glucose>galactose>mannose approximately fucose. Time course analyses of CO(2) production provided evidence of significant isotopic dilution; although pore water glucose turnover times were on the order of minutes, CO(2) did not plateau until after approximately 6 h of incubation. At this time a maximum of about 40% of the added radioglucose had been respired. The extent of respiration varied as a function of sediment depth and season, with the highest values below the surface (4 to 7 cm) and in summer and fall. Incorporation of radiolabelled glucose into biomass also varied seasonally, but the greatest extent of incorporation (about 40%) was observed in the fall and for the 0- to 1-cm depth interval. The production of short-chain fatty acid end products was largely limited to acetate, which accounted for only a small percentage of the added radiolabel. Other organic acids, pyruvate in particular, were observed in pore water and were due to artifacts in the heat-kill procedure used to terminate incubations. An accurate assessment of the distribution and importance of short-chain fatty acids as end products required the use of an enzymatic technique coupled with high-pressure liquid chromatography to verify qualitative identities. PMID- 16348838 TI - Inoculation of woody legumes with selected arbuscular mycorrhizal fungi and rhizobia to recover desertified mediterranean ecosystems. AB - Revegetation strategies, either for reclamation or for rehabilitation, are being used to recover desertified ecosystems. Woody legumes are recognized as species that are useful for revegetation of water-deficient, low-nutrient environments because of their ability to form symbiotic associations with rhizobial bacteria and mycorrhizal fungi, which improve nutrient acquisition and help plants to become established and cope with stress situations. A range of woody legumes used in revegetation programs, particularly in Mediterranean regions, were assayed. These legumes included both exotic and native species and were used in a test of a desertified semiarid ecosystem in southeast Spain. Screening for the appropriate plant species-microsymbiont combinations was performed previously, and a simple procedure to produce plantlets with optimized mycorrhizal and nodulated status was developed. The results of a 4-year trial showed that (i) only the native shrub legumes were able to become established under the local environmental conditions (hence, a reclamation strategy is recommended) and (ii) biotechnological manipulation of the seedlings to be used for revegetation (by inoculation with selected rhizobia and mycorrhizal fungi) improved outplanting performance, plant survival, and biomass development. PMID- 16348839 TI - Occurrence of UV-Absorbing, Mycosporine-Like Compounds among Cyanobacterial Isolates and an Estimate of Their Screening Capacity. AB - A survey of 20 strains of cyanobacteria (belonging to 13 genera) isolated from habitats exposed to strong insolation revealed that 13 strains contained one or more water-soluble, UV-absorbing, mycosporine amino acid (MAA)-like compounds. Some of the compounds were identical in several strains. In all, 13 distinct compounds were found. The UV absorption spectra of MAAs complemented well that of the extracellular sunscreen pigment scytonemin, which many of the strains also produced. Even though the specific MAA contents were variable among strains, they were invariably higher when the cultures were grown with UV radiation than when it was absent. In five strains tested, the MAA complement accumulated as a solute in the cytoplasmic cell fraction. The sunscreen capacities of MAA and scytonemin and their combined capacity were estimated for each strain and condition on the basis of the specific contents, cell size, and cellular location of the compounds. The estimates suggested that significant, albeit not complete, protection from UV photodamage could be gained from the possession of either MAA or scytonemin but especially from simultaneous screening by both types of compounds. PMID- 16348840 TI - Evidence Regarding the UV Sunscreen Role of a Mycosporine-Like Compound in the Cyanobacterium Gloeocapsa sp. AB - The UV sunscreen role commonly ascribed to mycosporine-like amino acids (MAAs) was investigated with an isolate of the terrestrial cyanobacterium Gloeocapsa sp. strain C-90-Cal-G.(2), which accumulates intracellularly an MAA with absorbance maximum at 326 nm but produces no extracellular sunscreen compound (i.e., scytonemin). The intracellular concentrations of MAA achieved were directly related to the intensity of the UV radiation (maximum at 320 nm) received by the cells. However, the presence of high concentrations of MAA was not necessary for the physiological acclimation of the cultures to UV radiation. The measured sunscreen factor due to MAA in single cells was 0.3 (the MAA prevented 3 out of 10 photons from hitting potential cytoplasmic targets). High contents of MAA in the cells correlated with increased resistance to UV radiation. However, when resistance was gauged under conditions of desiccation, with inoperative physiological photoprotective and repair mechanisms, cells with high MAA specific contents were only 20 to 25% more resistant. Although UV radiation centered around both 320 and 365 nm resulted in chlorophyll a photobleaching and photoinhibition of photosynthesis, the difference in sensitivity correlated with MAA accumulation occurred only at 320 nm (absorbed by MAA) and not at 365 nm (not absorbed by MAA). This difference represents the maximal protection ascribable to the presence of MAA for single cells, i.e., if one does not consider the enhancing effects of colony formation on protection by sunscreens. PMID- 16348841 TI - Characterization of a Cell Envelope-Associated Proteinase Activity from Streptococcus thermophilus H-Strains. AB - The production and biochemical properties of cell envelope-associated proteinases from two strains of Streptococcus thermophilus (strains CNRZ 385 and CNRZ 703) were compared. No significant difference in proteinase activity was found for strain CNRZ 385 when cells were grown in skim milk medium and M17 broth. Strain CNRZ 703 exhibited a threefold-higher proteinase activity when cells were grown in low-heat skim milk medium than when grown in M17 broth. Forty-one percent of the total activity of CNRZ 385 was localized on the cell wall. The optimum pH for enzymatic activity at 37 degrees C was around 7.0. Serine proteinase inhibitors, such as phenylmethylsulfonyl fluoride and diisopropylfluorophosphate, inhibited the enzyme activity in both strains. The divalents cations Ca, Mg, and Mn were activators, while Zn and Cu were inhibitors. beta-Casein was hydrolyzed more rapidly than alpha(s1)-casein. The results of DNA hybridization and immunoblot studies suggested that the S. thermophilus cell wall proteinase and the lactococcal proteinase are not closely related. PMID- 16348842 TI - Restriction/Modification systems and restriction endonucleases are more effective on lactococcal bacteriophages that have emerged recently in the dairy industry. AB - Recently, eight lytic small isometric-headed bacteriophages were isolated from cheese-manufacturing plants throughout North America. The eight phages were different, but all propagated on one strain, Lactococcus lactis NCK203. On the basis of DNA homology, they were classified in the P335 species. Digestion of their genomes in vitro with restriction enzymes resulted in an unusually high number of type II endonuclease sites compared with the more common lytic phages of the 936 (small isometric-headed) and c2 (prolate-headed) species. In vivo, the P335 phages were more sensitive to four distinct lactococcal restriction and modification (R/M) systems than phages belonging to the 936 and c2 species. A significant correlation was found between the number of restriction sites for endonucleases (purified from other bacterial genera) and the relative susceptibility of phages to lactococcal R/M systems. Comparisons among these three phage species indicate that the P335 species may have emerged most recently in the dairy industry. PMID- 16348843 TI - Lactic Acid production in a mixed-culture biofilm reactor. AB - Novel solid supports, consisting of polypropylene blended with various agricultural materials (pp composite), were evaluated as supports for pure- and mixed-culture continuous lactic acid fermentations in biofilm reactors. Streptomyces viridosporus T7A (ATCC 39115) was used to form a biofilm, and Lactobacillus casei subsp. rhamnosus (ATCC 11443) was used for lactic acid production. For mixed-culture fermentations, a 15-day continuous fermentation of S. viridosporus was performed initially to establish the biofilm. The culture medium was then inoculated with L. casei subsp. rhamnosus. For pure-culture fermentation, L. casei subsp. rhamnosus was inoculated directly into the reactors containing sterile pp composite chips. The biofilm reactors containing various pp composite chips were compared with a biofilm reactor containing pure polypropylene chips and with a reactor containing a suspension culture. Continuous fermentation was started, and each flow rate (0.06 to 1.92 ml/min) was held constant for 24 h; steady state was achieved after 10 h. Lactic acid production was determined throughout the 24-h period by high-performance liquid chromatography. Production rates that were two to five times faster than those of the suspension culture (control) were observed for the pure- and mixed-culture bioreactors. Both lactic acid production rates and lactic acid concentrations in the culture medium were consistently higher in mixed-culture than in pure-culture fermentations. Biofilm formation on the chips was detected at harvest by chip clumping and Gram staining. PMID- 16348844 TI - Differentiation of Two Abortive Mechanisms by Using Monoclonal Antibodies Directed toward Lactococcal Bacteriophage Capsid Proteins. AB - Monoclonal antibodies were used to monitor the accumulation of the major capsid protein of the lactococcal small isometric bacteriophage u136 (P335 species) over the course of a one-step growth curve. A sandwich enzyme-linked immunosorbent assay was then used to distinguish two abortive phage resistance mechanisms, Hsp and Prf. Capsid protein production of u136 was almost totally inhibited by the Hsp-induced abortive mechanism, supporting previous data that this mechanism blocks phage DNA replication. Prf-induced abortive infection only partially (50%) inhibited capsid protein production, suggesting that this mechanism targets some other point, perhaps within transcription or translation processes. The results confirmed that Hsp and Prf act at different targets in the phage lytic cycle. Use of monoclonal antibodies also demonstrated that production of the major capsid protein is a nonlimiting step in the lytic cycle of lactococcal phage u136. PMID- 16348845 TI - Insertion of Transposon Tn917 Derivatives into the Lactococcus lactis subsp. lactis Chromosome. AB - Two transposition vectors, pTV32 and pLTV1, containing transposon Tn917 derivatives TV32 and LTV1, respectively, were introduced into Lactococcus lactis subsp. lactis MG1614. It was found that pTV32 and pLTV1 replicate and that TV32 and LTV1 transpose in this strain. A protocol for production of a collection of Tn917 insertions in L. lactis subsp. lactis was developed. The physical locations of TV32 on the chromosomal SmaI fragments of 62 independent transpositions were established by pulsed-field gel electrophoresis. These transpositions could be divided into at least 38 different groups that exhibited no Tn917-dominating hot spots on the L. lactis subsp. lactis chromosome. A total of 10 of the 62 transpositions resulted in strains that express beta-galactosidase. This indicates that there was fusion of the promoterless lacZ of the Tn917 derivatives to a chromosomal promoter. Thus, the Tn917-derived transposons should be powerful genetic tools for studying L. lactis subsp. lactis. PMID- 16348846 TI - Production of (R)-3-Chloro-1,2-Propanediol from Prochiral 1,3-Dichloro-2-Propanol by Corynebacterium sp. Strain N-1074. AB - The production of (R)-3-chloro-1,2-propanediol [(R)-MCP] from prochiral 1,3 dichloro-2-propanol (DCP) was examined with a bacterial strain identified as a Corynebacterium strain. The addition of glycerol as a carbon source or some chlorinated alcohols to a medium was effective for the induction of activity catalyzing the transformation of DCP into MCP. The optimum pH for (R)-MCP production by the resting cell reaction was around 8.0. The optical purity of (R) MCP formed was improved by keeping the level of DCP in the reaction mixture at a low concentration. (R)-MCP was obtained from 77.5 mM DCP with a 97.3% molar conversion yield and an 83.8% enantiomeric excess of its optical purity by periodic feeding of the substrate. PMID- 16348847 TI - Characterization of Ethanol Production from Xylose and Xylitol by a Cell-Free Pachysolen tannophilus System. AB - Whole cells and a cell extract of Pachysolen tannophilus converted xylose to xylitol, ethanol, and CO(2). The whole-cell system converted xylitol slowly to CO(2) and little ethanol was produced, whereas the cell-free system converted xylitol quantitatively to ethanol (1.64 mol of ethanol per mol of xylitol) and CO(2). The supernatant solution from high-speed centrifugation (100,000 x g) of the extract converted xylose to ethanol, but did not metabolize xylitol unless a membrane fraction and oxygen were also present. Fractionation of the crude cell extract by gel filtration resulted in an inactive fraction in which ethanol production from xylitol was fully restored by the addition of NAD and ADP. The continued conversion of xylose to xylitol in the presence of fluorocitrate, which inhibited aconitase, demonstrated that the tricarboxylic acid cycle was not the source of the electrons for the production of xylitol from xylose. Therefore, the source of the electrons is indirectly identified as an oxidative pentose-hexose cycle. PMID- 16348848 TI - Production of Extracellular Proteins by the Biocontrol Fungus Gliocladium virens. AB - Gliocladium virens is a common saprophytic fungus that is mycoparasitic on a large number of fungi. Responses of G. virens toward its environment were examined by monitoring the presence of extracellular proteins in culture fluid during time course experiments. Culture fluid of G. virens grown on glucose, washed cell walls of Rhizoctonia solani (one of its hosts), olive oil, or chitin contained beta-glucanase, N-acetylglucosaminidase, lipase, and proteinase activities. There were relatively minor amounts of other enzymatic activities tested. Levels of extracellular enzyme activity varied with the age of the culture and the substrate used as the carbon source. Substrate-associated differences in enzyme activities were detected as early as 8 h after transfer of mycelia from stationary-phase cultures to fresh media. When G. virens was grown on host cell wall material, beta-glucanase had the greatest specific activity of any enzyme tested at 8 h. This result suggests that beta-glucanase may be the first enzyme important in the G. virens-R. solani interaction. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that some of the polypeptides were present in the culture fluid at relatively constant amounts and others accumulated early, at intermediate times, or late in the 8-day incubation test period. Several of the polypeptides present in the culture fluid during the first 24 h disappeared completely by 48 h. Consequently, it appears that extracellular proteins in cultures of G. virens are regulated by a combination of gene regulation and protein degradation. PMID- 16348849 TI - Evaluation of the Biolog Substrate Utilization System To Identify and Assess Metabolic Variation among Strains of Xanthomonas campestris pv. Citri. AB - Metabolic fingerprints of 148 strains of Xanthomonas campestris pv. citri originating from 24 countries and associated with various forms of citrus bacterial canker disease (CBCD) were obtained by using the Biolog substrate utilization system. Metabolic profiles were used to attempt strain identification. Only 6.8% of the studied strains were correctly identified when the commercial Microlog 2N data base was used alone. When the data base was supplemented with data from 54 strains of X. campestris pv. citri (40 CBCD-A strains, 8 CBCD-B strains, and 6 CBCD-C strains) and data from 43 strains of X. campestris associated with citrus bacterial spot disease, the percentage of correct identifications was 70%. Thus, it is recommended that users supplement the commercial data base with additional data prior to using the program for identification purposes. The utilization of Tween 40 in conjunction with other tests can help to differentiate strains associated with CBCD and citrus bacterial spot disease. These results confirmed the separation of X. campestris pv. citri into different subgroups (strains associated with Asiatic citrus canker [CBCD-A], cancrosis B [CBCD-B], and Mexican lime canker [CBCD-C]). The utilization of l fucose, d-galactose, and alaninamide can be used as markers to differentiate strains associated with these groups. A single strain associated with bacteriosis of Mexican lime in Mexico (CBCD-D) was closely similar to CBCD-B strains. PMID- 16348850 TI - Manganese Peroxidase, Produced by Trametes versicolor during Pulp Bleaching, Demethylates and Delignifies Kraft Pulp. AB - Previous work has shown that Trametes (Coriolus) versicolor bleaches kraft pulp brownstock with the concomitant release of methanol. In this work, the fungus is shown to produce both laccase and manganese peroxidase (MnP) but not lignin peroxidase during pulp bleaching. MnP production was enhanced by the presence of pulp and/or Mn(II) ions. The maximum level of secreted MnP was coincident with the maximum rate of fungal bleaching. Culture filtrates isolated from bleaching cultures produced Mn(II)- and hydrogen peroxide-dependent pulp demethylation and delignification. Laccase and MnP were separated by ion-exchange chromatography. Purified MnP alone produced most of the demethylation and delignification exhibited by the culture filtrates. On the basis of the methanol released and the total and phenolic methoxyl contents of the pulp, it appears that MnP shows a preference for the oxidation of phenolic lignin substructures. The extensive increase in brightness observed in the fungus-treated pulp was not found with MnP alone. Therefore, either the MnP effect must be optimized or other enzymes or compounds from the fungus are also required for brightening. PMID- 16348851 TI - Kraft Pulp Bleaching and Delignification by Dikaryons and Monokaryons of Trametes versicolor. AB - The ability of 10 dikaryotic and 20 monokaryotic strains of Trametes (Coriolus) versicolor to bleach and delignify hardwood and softwood kraft pulps was assessed. A dikaryon (52P) and two of its mating-compatible monokaryons (52J and 52D) derived via protoplasting were compared. All three regularly bleached hardwood kraft pulp more than 20 brightness points (International Standards Organization) in 5 days and softwood kraft pulp the same amount in 12 days. Delignification (kappa number reduction) by the dikaryon and the monokaryons was similar, but the growth of the monokaryons was slower. Insoluble dark pigments were commonly found in the mycelium, medium, and pulp of the dikaryon only. Laccase and manganese peroxidase (MnP) but not lignin peroxidase activities were secreted during bleaching by all three strains. Their laccase and MnP isozyme patterns were compared on native gels. No segregation of isozyme bands between the monokaryons was found. Hardwood kraft pulp appeared to adsorb several laccase isozyme bands. One MnP isozyme (pI, 3.2) was secreted in the presence of pulp by all three strains, but a second (pI, 4.9) was produced only by 52P. A lower level of soluble MnP activity in one monokaryon (52D) was associated with reduced bleaching ability and a lower level of methanol production. Since monokaryon 52J bleached pulp better than its parent dikaryon 52P, especially per unit of biomass, this genetically simpler monokaryon will be the preferred subject for further genetic manipulation and improvement of fungal pulp biological bleaching. PMID- 16348852 TI - Formation and Regeneration of Methanococcus voltae Protoplasts. AB - Methanococcus voltae cells were converted into protoplasts by suspension in anaerobic 0.1 M Tris-HCl buffer containing 0.4 M sucrose and 0.05 M NaCl as osmoprotectants. Protoplast formation was monitored microscopically by observing the conversion of the typical irregularly shaped (uneven peripheries) coccoid whole cells to rounded forms with smooth peripheries. Although the procedure resulted in about 50% lysis of the initial number of cells, the remainder were converted to the rounded form. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electron microscopy of negatively stained cell preparations indicated that the treatment removed the wall layer from whole cells to yield protoplasts. Protoplast regeneration was evaluated by using optimized plating conditions and an anaerobic microplating technique. Between 50 and 63% of the initial number of protoplasts regenerated as colonies on agar medium (35 degrees C, 7 days). The colony and cell morphologies of the regenerated protoplasts were indistinguishable from those of whole cells plated under identical conditions. PMID- 16348853 TI - Stereospecificity of microbial hydrations of oleic Acid to 10-hydroxystearic Acid. AB - We recently described a simple method for ascertaining the stereochemical purities of hydroxy fatty acids (S. H. El-Sharkawy, W. Yang, L. Dostal, and J. P. N. Rosazza, Appl. Environ. Microbiol. 58:2116-2122, 1992) based on the H-nuclear magnetic resonance spectral analysis of diastereomeric S-(+)-O-acetylmandelate esters of hydroxystearates. This report describes the stereochemistries of microbial hydrations of oleic acid to 10-hydroxystearic acid by Nocardia aurantia (also known as Rhodococcus rhodochrous) ATCC 12674, Nocardia restrictus ATCC 14887, Mycobacterium fortuitum UI-53387, Pseudomonas species strain NRRL-2994, Pseudomonas species strain NRRL B-3266, and baker's yeast. 10(R)-hydroxystearic acid isolated from Pseudomonas species strain NRRL-2994 was the standard for use in the H-nuclear magnetic resonance spectral technique to permit simple assignments of the absolute configurations of 10-hydroxystearic acid produced by different microorganisms. While the R. rhodochrous ATCC 12674-mediated hydration of oleic acid gave mixtures of enantiomers 10(R)-hydroxystearic acid and 10(S) hydroxystearic acid, Pseudomonas species strain NRRL-B-3266 produced optically pure 10(R)-hydroxystearic acid. The remaining microorganisms stereoselectively hydrated oleic acid to 10(R)-hydroxystearic acid containing between 2 and 18% of the contaminating 10(S)-hydroxystearic acid. PMID- 16348854 TI - Biological Control of Agrobacterium tumefaciens, Colonization, and pAgK84 Transfer with Agrobacterium radiobacter K84 and the Tra Mutant Strain K1026. AB - The efficacies of Agrobacterium radiobacter K84 and K1026 in root colonization, crown gall control, and plasmid transfer were compared. Levels of root colonization by K84 and K1026 of Montclar and Nemaguard peach seedlings were similar during the 21 days of the experiment. Four strains of A. tumefaciens bv. 1 were used for soil inoculations in biological control experiments on GF677 and Adafuel peach x almond rootstocks; two were sensitive and two were resistant to agrocin 84. Both strains K84 and K1026 were very efficient in controlling the sensitive strains, but some tumors appeared with both treatments. In the biocontrol of resistant strains, no galls were observed in K1026-treated plants, but some K84-treated plants had galls. Recovery of agrobacteria from galls in experiments with sensitive and resistant strains showed that all of the isolates from the controls or K1026-treated plants and most of the isolates from K84 treated plants had the same characteristics as the inoculated strains. Nine isolates from the K84-treated plants growing in soil inoculated with one resistant strain were virulent and produced agrocin 84. These isolates had a plasmid that hybridized with a probe prepared with the BamHI C fragment from pAgK84. These results show the efficiency of K1026 in biocontrol of agrocin 84 sensitive and -resistant strains of A. tumefaciens and suggest the use of K1026 as a safer organism than K84 for biological control of crown gall. PMID- 16348855 TI - Strains of Corynebacterium glutamicum with Different Lysine Productivities May Have Different Lysine Excretion Systems. AB - The lysine excretion systems of three different lysine-producing strains of Corynebacterium glutamicum were characterized in intact cells. Two strains (DG 52 5 and MH 20-22B) are lysine producers of different efficiency. They were bred by classical mutagenesis and have a feedback-resistant aspartate kinase. The third strain (KK 25) was constructed from the wild type by introducing the feedback resistant aspartate kinase gene of strain MH 20-22B into its genome. The three strains were shown to possess different excretion systems. Export in strain KK 25 is much slower than in the two mutants. The differences between the two lysine producing strains are more subtle. K(m) and V(max) are similar, but pH dependence and membrane potential dependence reveal differences in the intrinsic properties of the carrier system. PMID- 16348856 TI - Culture and Decontamination Methods Affecting Enumeration of Phages Infecting Bacteroides fragilis in Sewage. AB - [This corrects the article on p. 2670 in vol. 58.]. PMID- 16348858 TI - Stereospecific Hydroxylation of Indan by Escherichia coli Containing the Cloned Toluene Dioxygenase Genes from Pseudomonas putida F1. AB - [This corrects the article on p. 3407 in vol. 58.]. PMID- 16348857 TI - Bacterioplankton Community Structure and Dynamics after Large-Scale Release of Nonindigenous Bacteria as Revealed by Low-Molecular-Weight-RNA Analysis. AB - [This corrects the article on p. 3388 in vol. 58.]. PMID- 16348859 TI - Benthic Distribution of Sewage Sludge Indicated by Clostridium perfringens at a Deep-Ocean Dump Site. AB - Clostridium perfringens in sediment samples collected at the Deep Water Municipal Sewage Sludge Disposal Site (also called the 106-Mile Site), off the coast of New Jersey, was enumerated. The counts of C. perfringens found in sediment samples collected within and to the southwest of the 106-Mile Site were significantly elevated (P < 0.01) compared with counts of samples from reference stations of similar depth (2,400 to 2,700 m), topography, and distance from the continental shelf, indicating that the benthic environment was contaminated by sewage dumping at this site. Low counts of C. perfringens in sediment samples collected at stations between the base of the continental shelf and the 106-Mile Site indicated that coastal runoff was not a significant source of contamination. Elevated counts were observed for samples up to 92 km to the southwest, whereas low counts were obtained for samples from stations to the east of the 106-Mile Site. This distribution is consistent with previous model predictions of sludge deposition. In areas heavily impacted by sludge dumping, C. perfringens counts were generally highest in the top 1 cm of sediment and exceeded 9,000 CFU g (dry weight) of sediment. The patterns of C. perfringens dispersal observed in this study have proved useful for selection of heavily impacted areas and control stations for further ecological evaluation by a multidisciplinary research team. PMID- 16348860 TI - Antagonistic Effect of Nonpathogenic Fusarium oxysporum Fo47 and Pseudobactin 358 upon Pathogenic Fusarium oxysporum f. sp. dianthi. AB - Pseudobactin production by Pseudomonas putida WCS358 significantly improves biological control of fusarium wilt caused by nonpathogenic Fusarium oxysporum Fo47b10 (P. Lemanceau, P. A. H. M. Bakker, W. J. de Kogel, C. Alabouvette, and B. Schippers, Appl. Environ. Microbiol. 58:2978-2982, 1992). The antagonistic effect of Fo47b10 and purified pseudobactin 358 was studied by using an in vitro bioassay. This bioassay allows studies on interactions among nonpathogenic F. oxysporum Fo47b10, pathogenic F. oxysporum f. sp. dianthi WCS816, and purified pseudobactin 358, the fluorescent siderophore produced by P. putida WCS358. Both nonpathogenic and pathogenic F. oxysporum reduced each other's growth when grown together. However, in these coinoculation experiments, pathogenic F. oxysporum WCS816 was relatively more inhibited in its growth than nonpathogenic F. oxysporum Fo47b10. The antagonism of nonpathogenic F. oxysporum against pathogenic F. oxysporum strongly depends on the ratio of nonpathogenic to pathogenic F. oxysporum densities: the higher this ratio, the stronger the antagonism. This fungal antagonism appears to be mainly associated with the competition for glucose. Pseudobactin 358 reduced the growth of both F. oxysporum strains, whereas ferric pseudobactin 358 did not; antagonism by pseudobactin 358 was then related to competition for iron. However, the pathogenic F. oxysporum strain was more sensitive to this antagonism than the nonpathogenic strain. Pseudobactin 358 reduced the efficiency of glucose metabolism by the fungi. These results suggest that pseudobactin 358 increases the intensity of the antagonism of nonpathogenic F. oxysporum Fo47b10 against pathogenic F. oxysporum WCS816 by making WCS816 more sensitive to the glucose competition by Fo47b10. PMID- 16348861 TI - Effects of Calcium on Sugar Transport in Azotobacter vinelandii. AB - A fast and environmentally safe procedure was used to study sugar uptake by Azotobacter vinelandii. Transport experiments were performed in a 24-well plate and aerated by rapid oscillatory vibration. Samples were washed by centrifugation and dissolved in biodegradable scintillation cocktail for counting. At cell concentrations up to 6 x 10 cells per ml, the uptake of sucrose was a function of time and was proportional to the cell concentration. This modified uptake assay was used to test the effect of cations on sugar uptake in A. vinelandii. Results showed that Ca at 1 to 2 mM stimulated sucrose uptake by decreasing the apparent K(m) of sucrose transport. Higher Ca concentrations inhibited sucrose uptake in this organism. PMID- 16348862 TI - Organic thiols as organolithotrophic substrates for growth of phototrophic bacteria. AB - Rhodopseudomonas sp. strain BB1, isolated from a coastal marine sediment, immediately metabolized mercaptomalate when grown on mercaptomalate. Sulfide was detected as an intermediate. Extracts of cells grown on mercaptomalate converted mercaptomalate or 3-mercaptopropionate to equimolar amounts of sulfide and either fumarate or acrylate, respectively. Rhodopseudomonas sp. strain BB1 gave higher growth yields on mercaptomalate than on sulfide or malate, consistent with metabolism of the carbon chain of the thiol and the liberated sulfide; i.e., the organic thiol was an organolithotrophic substrate. In contrast, Thiocapsa roseopersicina, isolated previously from a marine microbial mat, had similar growth yields on sulfide, mercaptomalate, or 3-mercaptopropionate, with fumarate or acrylate accumulation from the thiols. T. roseopersicina did not grow photoorganotrophically on fumarate or acrylate, and the thiols were only a source of sulfide for photolithoautotrophic growth. PMID- 16348863 TI - The Mode of Replication Is a Major Factor in Segregational Plasmid Instability in Lactococcus lactis. AB - The effects of the rolling-circle and theta modes of replication on the maintenance of recombinant plasmids in Lactococcus lactis were studied. Heterologous Escherichia coli or bacteriophage lambda DNA fragments of various sizes were inserted into vectors based on either the rolling-circle-type plasmid pWV01 or the theta-type plasmid pAMbeta1. All pAMbeta1 derivatives were stably maintained. pWV01 derivatives, however, showed size-dependent segregational instability, in particular when large DNA fragments were inserted. All recombinant pWV01 derivatives generated high-molecular-weight plasmid multimers (HMW) in amounts that were positively correlated with plasmid size and inversely correlated with the copy numbers of the monomeric plasmid forms. Formation of HMW or reductions in copy numbers were not observed with pAMbeta1 derivatives. The results indicate that HMW formation and/or reduction in plasmid copy numbers is an important factor in the maintenance of pWV01 derivatives. It is concluded that theta-type plasmids are superior to rolling-circle-type plasmids for cloning in lactococci. PMID- 16348864 TI - A strategy for rotation of different bacteriophage defenses in a lactococcal single-strain starter culture system. AB - A new strategy for starter culture rotations was developed for a series of phage resistant clones genetically derived from a single strain of Lactococcus lactis subsp. lactis. Phage-resistant derivatives carrying different defense systems were constructed via conjugation with various plasmids encoding abortive infection (Abi/Hsp) and/or restriction and modification (R/M) systems of different specificity. The plasmids included pTR2030 (Hsp R/M), pTN20 (Abi R/M), pTRK11 (R/M), and pTRK68 (R/M). Selected phage-resistant transconjugants or transformants were evaluated in different rotation sequences through cycles of the Heap-Lawrence starter culture activity test in milk contaminated with phage and whey from the previous cycle. When used in consecutive sequence, derivative strains carrying the R/M systems encoded by pTN20, pTRK11, and pTRK68 retarded phage development when the initial levels of phage contamination were below 10 PFU/ml but not when levels were increased to 10 PFU/ml. Use of a derivative bearing pTR2030 (Hsp R/M) at the beginning of the rotation prevented phage development, even when the initial levels of phage contamination were high (10 PFU/ml). Alternating the type and specificity of R/M and Abi defenses through the rotation prevented phage proliferation and in some cases eliminated contaminating phages. A model rotation sequence for the phage defense rotation strategy was developed and performed successfully over nine cycles of the Heap-Lawrence starter culture activity test in the presence of high-titer commercial phage composites. This phage defense rotation strategy is designed to protect a highly specialized Lactococcus strain from phage attack during continuous and extended use in the dairy industry. PMID- 16348865 TI - Role of Chitin-Binding Proteins in the Specific Attachment of the Marine Bacterium Vibrio harveyi to Chitin. AB - We examined the mechanism of attachment of the marine bacterium Vibrio harveyi to chitin. Wheat germ agglutinin and chitinase bind to chitin and competitively inhibited the attachment of V. harveyi to chitin, but not to cellulose. Bovine serum albumin and cellulase do not bind to chitin and had no effect on bacterial attachment to chitin. These data suggest that this bacterium recognizes specific attachment sites on the chitin particle. The level of attachment of a chitinase overproducing mutant of V. harveyi to chitin was about twice as much as that of the uninduced wild type. Detergent-extracted cell membranes inhibited attachment and contained a 53-kDa peptide that was overproduced by the chitinase overproducing mutant. Three peptides (40, 53, and 150 kDa) were recovered from chitin which had been exposed to membrane extracts. Polyclonal antibodies raised against extracellular chitinase cross-reacted with the 53- and 150-kDa chitin binding peptides and inhibited attachment, probably by sterically hindering interactions between the chitin-binding peptides and chitin. The 53- and 150-kDa chitin-binding peptides did not have chitinase activity. These results suggest that chitin-binding peptides, especially the 53-kDa chitin-binding peptide and chitinase and perhaps the 150-kDa peptide, mediate the specific attachment of V. harveyi to chitin. PMID- 16348867 TI - Dissolved Nitrogen Uptake by a Cyanobacterial Bloom (Anabaena flos-aquae) in a Subarctic Lake. AB - Uptake of dissolved nitrogen (NH(4) + NO(3) + urea + N(2)) by a cyanobacterial [Anabaena flos-aquae (Lyngb.)] De Breb population in Smith Lake, Alaska, was measured every 2 to 4 days during the spring of 1990. Total dissolved nitrogen uptake ranged from 0.34 to 24.75 mumol liter h, with a mean of 5.75 mumol liter h; the euphotic zone accounted for 91% of the uptake. The mean turnover time for dissolved combined nitrogen (NH(4) + NO(3) + urea) in the euphotic zone was less than 14 h, and that for NH(4) was only 3.6 h. The mean relative preference indices for NH(4) (2.4), NO(3) (0.4), and urea (0.5) established NH(4) as the preferred nitrogenous nutrient. The uptake rates were apparently dependent on biomass, temperature, and light. Regeneration, probably due to zooplankton excretion and bacterial remineralization of dissolved organic nitrogen, was the main source of NH(4) for the cyanobacterial growth. The high half-saturation constant for NH(4) with low ambient NH(4) concentration nevertheless resulted in the simultaneous utilization of several forms of nitrogen. PMID- 16348866 TI - Environmental factors affecting the occurrence of mycobacteria in brook waters. AB - To evaluate the impact of environmental factors on the occurrence of environmental mycobacteria, viable counts of mycobacteria were measured in samples of brook water collected from 53 drainage areas located in a linear belt crossing Finland at 63 degrees north latitude. The numbers of mycobacteria were correlated with characteristics of the drainage area, climatic parameters, chemical and physical characteristics of the water, and counts of other heterotrophic bacteria in the water. The numbers of mycobacteria in the water ranged from 10 to 2,200 CFU/liter. The counts correlated positively (P < 0.001) with the presence of peatlands, precipitation data, chemical oxygen demand, water color, and concentrations of Fe, Al, Cu, Co, and Cr. The mycobacterial counts correlated negatively (P < 0.001) with water pH, whereas other heterotrophic bacterial counts lacked any correlation with pH. A linear regression model with four independent variables (i.e., peatlands in the drainage area, chemical oxygen demand, concentration of potassium, and pH) explained 83% of the variation in mycobacterial counts in brook waters. Our results suggest that acidification may enhance the growth of environmental mycobacteria. PMID- 16348868 TI - Microgradients of microbial oxygen consumption in a barley rhizosphere model system. AB - A microelectrode technique was used to map the radial distribution of oxygen concentrations and oxygen consumption rates around single roots of 7-day-old barley seedlings. The seedlings were grown in gel-stabilized medium containing a nutrient solution, a soil extract, and an inert polymer. Oxygen consumption by microbial respiration in the rhizosphere (<5 mm from the root) and in bulk medium (>30 mm from the root) was determined by using Fick's laws of diffusion and an analytical approach with curve fitting to measured microprofiles of oxygen concentration. A marked increase of microbial respiration was observed in the inner 0- to 3-mm-thick, concentric zone around the root (rhizosphere). The volume specific oxygen consumption rate (specific activity) was thus 30 to 60 times higher in the innermost 0 to 0.01 mm (rhizoplane) than in the bulk medium. The oxygen consumption rate in the root tissue was in turn 10 to 30 times higher than that in the rhizoplane. Both microbial respiration and oxygen uptake by the root varied between different roots. This was probably due to a between-root variation of the exudation rate for easily degradable carbon compounds supporting the microbial oxygen consumption. PMID- 16348869 TI - Xylosylation of Phenolic Hydroxyl Groups of the Monomeric Lignin Model Compounds 4-Methylguaiacol and Vanillyl Alcohol by Coriolus versicolor. AB - When 4-methylguaiacol (MeG), a phenolic lignin model compound, was added to a culture that was inoculated with Coriolus versicolor, it was bioconverted into 2 methoxy-4-methylphenyl beta-d-xyloside (MeG-Xyl). The phenolic hydroxyl group of vanillyl alcohol was much more extensively xylosylated than the alcoholic hydroxyl group. When a mixture of MeG and commercial UDP-xylose was incubated with cell extracts of mycelia, transformation of UDP-xylose into MeG-Xyl was observed. This result suggested that UDP-xylosyltransferase was involved in the xylosylation of phenolic hydroxyl groups of lignin model compounds. PMID- 16348870 TI - Gene expression per gene dose, a specific measure of gene expression in aquatic microorganisms. AB - A method for the measurement of specific levels of gene expression that combines target mRNA and target DNA quantitation has been developed. The use of target gene dose as a normalizing factor for mRNA provides an alternative to 16S or 23S rRNA, which are unsuitable for use in the environment because of their presence in nontarget organisms. Both target mRNA and DNA are recovered from replicate samples and detected by using antisense and sense single-stranded RNA gene probes. For efficient mRNA recovery, the use of Millipore Durapore filters and multiple extractions was necessary. Quantitation was performed by radiometric detection by using a beta-scanner and comparison of the sample signal against target mRNA and DNA standard curves. This method enabled the measurement of expression of the catechol-2,3-dioxygenase gene (xylE) contained on the thermoregulated plasmid pLV1013 in a marine Vibrio strain in culture and in the environment. In studies of the relationship between mRNA levels and enzyme activities, the appearance of enzyme activity lagged behind xylE mRNA synthesis by an hour after temperature induction. This suggests that mRNA analysis is well suited for determining rapid regulation of microbial gene expression at the transcriptional level in water column microbial populations. PMID- 16348871 TI - Protozoa in subsurface sediments from sites contaminated with aviation gasoline or jet fuel. AB - Numbers of protozoa in the subsurface of aviation gasoline and jet fuel spill areas at a Coast Guard base at Traverse City, Mich., were determined. Boreholes were drilled in an uncontaminated location, in contaminated but untreated parts of the fuel plumes, and in the aviation gasoline source area undergoing H(2)O(2) biotreatment. Samples were taken from the unsaturated zone to depths slightly below the floating free product in the saturated zone. Protozoa were found to occur in elevated numbers in the unsaturated zone, where fuel vapors mixed with atmospheric oxygen, and below the layer of floating fuel, where uncontaminated groundwater came into contact with fuel. The same trends were noted in the biotreatment area, except that numbers of protozoa were higher. Numbers of protozoa in some contaminated areas equalled or exceeded those found in surface soil. The abundance of protozoa in the biotreatment area was high enough that it would be expected to significantly reduce the bacterial community that was degrading the fuel. Little reduction in hydraulic conductivity was observed, and no bacterial fouling of the aquifer was observed during biotreatment. PMID- 16348872 TI - Methane consumption in temperate and subarctic forest soils: rates, vertical zonation, and responses to water and nitrogen. AB - Rates of methane consumption were measured in subarctic coniferous and temperate mixed-hardwood forest soils, using static chambers and intact soil cores. Rates at both sites were generally between 1 and 3 mg of CH(4) m day and decreased with increasing soil water contents above 20%. Addition of ammonium (1 mumol g of soil) strongly inhibited methane oxidation in the subarctic soils; a lesser inhibition was observed for temperate forest samples. The response to nitrogen additions occurred within a few hours and was probably due to physiological changes in the active methane-consuming populations. Methane consumption in soils from both sites was stratified vertically, with a pronounced subsurface maximum. This maximum was coincident with low levels of both nitrate and ammonium in the mixed-hardwood forest soil. PMID- 16348873 TI - Regulation of the Periplasmic [Fe] Hydrogenase by Ferrous Iron in Desulfovibrio vulgaris (Hildenborough). AB - The periplasmic [Fe] hydrogenase from the sulfate-reducing bacterium Desulfovibrio vulgaris (Hildenborough) DSM 8303 was found to be regulated by ferrous iron availability. During growth with 5 ppm of iron, the enzyme derepressed and the specific activity increased approximately fourfold, whereas the presence of 100 ppm of ferrous iron repressed the enzyme. The repression derepression phenomenon with ferrous iron was found to be operative when the cells were cultured under either hydrogen or nitrogen gas. This is the first reported case showing that the hydrogenase enzyme is regulated by iron, and the implications of this finding relative to the corrosion industry are discussed. PMID- 16348874 TI - Ergosterol-to-Biomass Conversion Factors for Aquatic Hyphomycetes. AB - Fourteen strains of aquatic hyphomycete species that are common on decaying leaves in running waters were grown in liquid culture and analyzed for total ergosterol contents. Media included an aqueous extract from senescent alder leaves, a malt extract broth, and a glucose-mineral salt solution. Concentrations of ergosterol in fungal mycelium ranged from 2.3 to 11.5 mg/g of dry mass. The overall average was 5.5 mg/g. Differences among both species and growth media were highly significant but followed no systematic pattern. Stationary-phase mycelium had ergosterol contents 10 to 12% lower or higher than mycelium harvested during the growth phase, but these differences were only significant for one of four species examined. Availability of plant sterols in the growth medium had no clear effect on ergosterol concentrations in two species tested. To convert ergosterol contents determined in field samples to biomass values of aquatic hyphomycetes, a general multiplicative factor of 182 is proposed. More accurate estimates would be obtained with species-specific factors. Using these in combination with estimates of the proportion of the dominant species in a naturally established community on leaves resulted in biomass estimates that were typically 20% lower than those obtained with the general conversion factor. Improvements of estimates with species-specific factors may be limited, however, by intraspecific variability in fungal ergosterol content. PMID- 16348875 TI - Microelectrode measurements of the activity distribution in nitrifying bacterial aggregates. AB - Microelectrodes for ammonium, oxygen, nitrate, and pH were used to study nitrifying aggregates grown in a fluidized-bed reactor. Local reactant fluxes and distribution of microbial activity could be determined from the microprofiles. The interfacial fluxes of the reactants closely reflected the stoichiometry of bacterial nitrification. Both ammonium consumption and nitrate production were localized in the outer shells, with a thickness of approximately 100 to 120 mum, of the aggregates. Under conditions in which ammonium and oxygen penetrated the whole aggregate, nitrification was restricted to this zone; oxygen was consumed in the central parts of the aggregates as well, probably because of oxidation of dead biomass. A sudden increase of the oxygen concentration to saturation (pure oxygen) was inhibitory to nitrification. The pH profiles showed acidification in the aggregates, but not to an inhibitory level. The distribution of activity was determined by the penetration depth of oxygen during aggregate development in the reactor. Mass transfer was significantly limited by the boundary layer surrounding the aggregates. Microelectrode measurements showed that the thickness of this layer was correlated with the diffusion coefficient of the species. Determination of the distribution of nitrifying activity required the use of ammonium or nitrate microelectrodes, whereas the use of oxygen microelectrodes alone would lead to erroneous results. PMID- 16348876 TI - Karyotypic Variation within Clonal Lineages of the Rice Blast Fungus, Magnaporthe grisea. AB - We have analyzed the karyotype of the rice blast fungus, Magnaporthe grisea, by using pulsed-filed gel electrophoresis. We tested whether the electrophoretic karyotype of an isolate was related to its pathotype, as determined by infection assays, or its genetic lineage, as determined by DNA fingerprinting. Highly reproducible electrophoretic karyotypes were obtained for a collection of U.S. and Chinese isolates representing a diverse collection of pathotypes and genetic lineages. Chromosomes ranged in size from 3 to 10 Mb. Although chromosome number was largely invariant, chromosome length polymorphisms were frequent. Minichromosomes were also found, although their presence was not ubiquitous. They ranged in number from 1 to 3 and in size from 470 kb to 2.2 Mb. Karyotypes were sufficiently variable as to obscure the obvious relatedness of isolates on the basis of pathogenicity assays or genetic lineage analysis by DNA fingerprinting. We documented that the electrophoretic karyotype of an isolate can change after prolonged serial transfer in culture and that this change did not alter the isolate's pathotype. The mechanisms bringing about karyotype variability involve deletions, translocations, and more complex rearrangements. We conclude that karyotypic variability in the rice blast fungus is a reflection of the lack of sexuality in wild populations which leads to the maintenance of neutral genomic rearrangements in clones of the fungus. PMID- 16348877 TI - Fluorinated vitamin b(12) analogs are cofactors of corrinoid-dependent enzymes: a f-labeled nuclear magnetic resonance probe for identifying corrinoid-protein interactions. AB - The homoacetogenic bacterium Sporomusa ovata synthesized the vitamin B(12) analog phenolyl cobamide or 4-fluorophenolyl cobamide when the methanol medium of growing cells was supplemented with 10 mM phenol or 5 mM 4-fluorophenol. Phenol and, presumably, 4-fluorophenol were specifically incorporated into these cobamides, since phenol was not metabolized significantly into amino acids or into acetic acid, the product of the catabolism. The phenol-containing cobamides contributed up to 90% of the protein-bound cobamides of the 1,300 to 1,900 nmol of corrinoid per g of dry cell material formed. Fluorine-19 nuclear magnetic resonance spectroscopy of 4-fluorophenolyl cobamide exhibited a resonance near 30 ppm. An additional signal emerged at 25 ppm when 4-fluorophenolyl cobamide was investigated as the cofactor of a corrinoid-dependent protein. The two resonances indicated distinct cofactor arrangements within the protein's active site. A 5 ppm high-field shift change suggested van der Waal's interactions between the fluorinated nucleotide of the cofactor and adjacent amino acid residues of the enzyme. Similarly, Propionibacterium freudenreichii and Methanobacterium thermoautotrophicum synthesized 5-fluorobenzimidazolyl cobamide. The human corrinoid binders intrinsic factor, transcobalamin, and haptocorrin recognized this corrinoid like vitamin B(12). Hence, it is possible to use F-labeled nuclear magnetic resonance spectroscopy for analyses of protein-bound cobamides. PMID- 16348878 TI - On-Line Detection of Substrate Exhaustion by Using NAD(P)H Fluorescence. AB - In this study we examined the utility of NAD(P)H fluorescence for monitoring aerobic fermentations of the threonine auxotroph Corynebacterium glutamicum ATCC 14296. Instead of attempting complicated mathematical corrections for inner filter effects, we found that it is possible to use the information contained in the on-line NAD(P)H fluorescence signal to assess culture metabolic activities during fermentation. The first derivative of the filtered fluorescence signal, which approximates the turnover rate of the NAD(P)H pool, can be used to precisely identify the temporal points of threonine and glucose exhaustion. PMID- 16348880 TI - Inhibition of Butyrate Oxidation by Formate during Methanogenesis. AB - A mixed methanogenic culture fed with glucose was perturbed with butyrate and formate to investigate the role of formate in the acetogenesis of butyrate. A free energy analysis suggests that formate rather than H(2) was the interspecies electron carrier for butyrate conversion into acetate for the culture studied. PMID- 16348879 TI - Amino Acid Requirements of Two Hyperthermophilic Archaeal Isolates from Deep-Sea Vents, Desulfurococcus Strain SY and Pyrococcus Strain GB-D. AB - Two sulfur-dependent hyperthermophilic archaea, Desulfurococcus strain SY and Pyrococcus strain GB-D, which were isolated from deep-sea hydrothermal vents, utilized free amino acids and peptides obtained from various molecular size fractions of yeast extract. It was found that 11 amino acids were essential for growth. The metabolic products were acetate, i-butyrate, and i-valerate. PMID- 16348881 TI - Isolation of Rhizobium meliloti nod Gene Inducers from Alfalfa Rhizosphere Soil. AB - Methanolic extracts of alfalfa rhizosphere soil induce more nod gene transcription in Rhizobium meliloti than extracts of nonrhizosphere soil. Six peaks of nod-inducing activity were separated by high-performance liquid chromatography from rhizosphere soil extract, and one compound was identified by H nuclear magnetic resonance, mass spectrometry, and UV-visible spectra as a formononetin-7-O-glycoside that activates both NodD1 and NodD2 proteins. The unanticipated presence of a glycoside in rhizosphere soil suggests either that large amounts of the glycoside were exuded by roots or that some glycosides are unexpectedly stable in soil. PMID- 16348882 TI - Dipicolinic Acid Greatly Enhances Production of Spore Photoproduct in Bacterial Spores upon UV Irradiation. AB - Formation of the spore photoproduct (SP) (5-thyminyl-5,6-dihydrothymine) in DNA of dormant spores of Bacillus subtilis upon UV irradiation is due to binding of alpha/beta-type small, acid-soluble proteins (SASP). However, the yield of SP as a function of UV fluence is approximately 15-fold higher in spores than in an alpha/beta-type-SASP-DNA complex in vitro. The yield of SP as a function of UV fluence in forespore DNA from mutants which make alpha/beta-type SASP but not dipicolinic acid (DPA) was 10 to 20 times lower than that in dormant spores. Furthermore, the yield of SP as a function of UV fluence in an alpha/beta-type SASP-DNA complex in vitro was increased sixfold by DPA. These data provide further support for the idea that the high DPA level in dormant spores increases the yield of SP as a function of UV fluence and thereby sensitizes spores to UV. PMID- 16348883 TI - Modification of the Fe Protein of Nitrogenase in Natural Populations of Trichodesmium thiebautii. AB - The Fe protein of nitrogenase in the marine nonheterocystous cyanobacterium Trichodesmium thiebautii is interconverted between two forms, which is reminiscent of the ADP-ribosylation described in the purple bacterium Rhodospirillum rubrum. In natural populations of T. thiebautii during the day, when nitrogenase activity (NA) is present and while photosynthetic rates are high, a low-molecular-mass form of the Fe protein is present. In the late afternoon, the low-molecular-mass form is partially converted to a higher molecular-mass form (approximately equal distribution of high- and low-molecular mass forms of the Fe protein subunits), concurrent with cessation of NA. Some of the higher-molecular-mass form persists through the night until the very early morning, when the lower-molecular-mass form appears. New synthesis of both the Fe and MoFe proteins of nitrogenase appears to occur at this time. The higher molecular-mass form of the Fe protein is also produced rapidly in response to artificially elevated external O(2) levels (40%) during the day. T. thiebautii is capable of recovery of NA in less than 1 h following exposure to 40% O(2), which is correlated with the return of the Fe protein to the lower-molecular-mass form. Recovery from exposure to O(2) is not dependent upon protein synthesis. The modification of the Fe protein is clearly involved in regulation of NA during the diel cycle of NA in T. thiebautii but may also be involved in protecting the Fe protein during transient O(2) concentration increases. PMID- 16348884 TI - A method for detection of pseudobactin, the siderophore produced by a plant growth-promoting pseudomonas strain, in the barley rhizosphere. AB - Detection in the rhizosphere of the siderophore produced by an inoculated microorganism is critical to determining the role of microbial iron chelators on plant growth promotion. We previously reported the development of monoclonal antibodies (MAb) to ferric pseudobactin, the siderophore of plant-growth promoting Pseudomonas strain B10. One of these MAb reacted less strongly to pseudobactin than to ferric pseudobactin. The MAb reacted to Al(III), Cr(III), Cu(II), and Mn(II) complexes of pseudobactin at a level similar to the level at which it reacted to ferric pseudobactin and reacted less to the Zn(II) complex, but these metals would make up only a small fraction of chelated pseudobactin in soil on the basis of relative abundance of metals and relative binding constants. Fourteen-day-old barley plants grown in limed and autoclaved soil were inoculated with 10 CFU of Pseudomonas strain Sm1-3, a strain of Pseudomonas B10 Rif Nal selected for enhanced colonization, and sampled 3 days later. Extraction and analysis of the roots and surrounding soil using the MAb in an immunoassay indicated a concentration of 3.5 x 10 mol of ferric pseudobacting g (wet weight). This is the first direct measurement of a pseudobactin siderophore in soil or rhizosphere samples. PMID- 16348885 TI - Transformation of 3- and 4-Picoline under Sulfate-Reducing Conditions. AB - A microbial population which transformed 3- and 4-picoline under sulfate-reducing conditions was isolated from a subsurface soil which had been previously exposed to different N-substituted aromatic compounds for several years. In the presence of sulfate, the microbial culture transformed 3- and 4-picoline (0.4 mM) within 30 days. From the amounts of ammonia released and of sulfide that were determined during the transformation of 3-picoline, it can be concluded that the parent compound was mineralized to carbon dioxide and ammonia. During the transformation of 4-picoline, a UV-absorbing intermediate accumulated in the culture medium. This metabolite was identified as 2-hydroxy-4-picoline by gas chromatography-mass spectrometry and nuclear magnetic resonance analysis, and its further transformation was detected only after an additional month of incubation. The small amount of sulfide produced during the oxidation of 4-picoline and the generation of the hydroxylated metabolite indicated that the initial step in the metabolic pathway of 4-picoline was a monohydroxylation at position 2 of the heterocyclic aromatic ring. The 3- and 4-picoline-degrading cultures could also transform benzoic acid; however, the other methylated pyridine derivatives, 2 picoline, dimethyl-pyridines, and trimethylpyridines, were not degraded. PMID- 16348886 TI - Purification and Properties of an S-Adenosylmethionine: 2,4-Disubstituted Phenol O-Methyltransferase from Phanerochaete chrysosporium. AB - An enzyme catalyzing the O-methylation of acetovanillone (3-methoxy-4 hydroxyacetophenone) by S-adeno-sylmethionine was isolated from Phanerochaete chrysosporium and purified 270-fold by ultrafiltration, anion-exchange chromatography, and gel filtration. The enzyme exhibited a pH optimum between 7 and 9 and was rapidly denatured at temperatures above 55 degrees C. The K(m) values for acetovanillone and S-adenosylmethionine were 34 and 99 muM, respectively. S-Adenosylhomocysteine acted as a powerful competitive inhibitor of S-adenosylmethionine, with a K(i) of 41 muM. The enzyme was also susceptible to inhibition by thiol reagents and low concentrations of heavy metal ions. Gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the enzyme was monomeric and had a molecular weight of approximately 53,000. Substrate specificity studies showed that 3-methoxy- and 3,5-dimethoxy-substituted 4-hydroxy-benzaldehydes, -benzoic acids, and acetophenones were the preferred substrates for the enzyme. The corresponding 3,4 dihydroxy compounds were methylated relatively slowly, while the 3-hydroxy-4 methoxy compounds were almost inactive as substrates. Substituents in both the 2 and 4 positions relative to the hydroxyl group appeared to be essential for significant enzyme attack of a substrate. Provided that certain steric criteria were satisfied, the nature of the substituent was not critical. Hence, xenobiotic compounds such as 2,4-dichlorophenol and 2,4-dibromophenol were methylated almost as readily as acetovanillone. However, an extended side chain in the 4 position was not compatible with activity as a substrate, and neither homovanillic, caffeic, nor ferulic acid was methylated. The substrate range of the O methyltransferase tends to imply a role in the catabolism or detoxification of lignin degradation products such as vanillic and syringic acids. PMID- 16348887 TI - Abundance of Virus-Sized Non-DNase-Digestible DNA (Coated DNA) in Eutrophic Seawater. AB - Total DNA concentration in 0.2-mum-pore-size Nuclepore filter filtrates (<0.2-mum fraction) of Tokyo Bay water was estimated to be 9 to 19 ng/ml by an immunochemical quantification method. Almost 90% of the DNA in the <0.2-mum fraction was found in the size fractions larger than 3.0 x 10 Da and 0.03 mum, and most was not susceptible to DNase digestion, that is, consisted of non-DNase digestible DNA (coated DNA). A significant amount of DNA was obtained from the <0.2-mum fraction of the seawater by three different methods: polyethylene glycol precipitation, direct ethanol precipitation, and ultrafilter concentration. Gel electrophoresis analysis of the isolated DNAs showed that they consisted mainly of coated DNAs with a similar molecular sizes (20 to 30 kb [1.3 x 10 to 2.0 x 10 Da). The abundance of the ultramicron virus-sized coated DNA in natural seawater suggests that these DNA-rich particles can be attributed to marine DNA virus assemblages and that they may be a significant phosphorus reservoir in the environment. PMID- 16348888 TI - Dissimilatory Fe(III) Reduction by the Marine Microorganism Desulfuromonas acetoxidans. AB - The ability of the marine microorganism Desulfuromonas acetoxidans to reduce Fe(III) was investigated because of its close phylogenetic relationship with the freshwater dissimilatory Fe(III) reducer Geobacter metallireducens. Washed cell suspensions of the type strain of D. acetoxidans reduced soluble Fe(III)-citrate and Fe(III) complexed with nitriloacetic acid. The c-type cytochrome(s) of D. acetoxidans was oxidized by Fe(III)-citrate and Mn(IV)-oxalate, as well as by two electron acceptors known to support growth, colloidal sulfur and malate. D. acetoxidans grew in defined anoxic, bicarbonate-buffered medium with acetate as the sole electron donor and poorly crystalline Fe(III) or Mn(IV) as the sole electron acceptor. Magnetite (Fe(3)O(4)) and siderite (FeCO(3)) were the major end products of Fe(III) reduction, whereas rhodochrosite (MnCO(3)) was the end product of Mn(IV) reduction. Ethanol, propanol, pyruvate, and butanol also served as electron donors for Fe(III) reduction. In contrast to D. acetoxidans, G. metallireducens could only grow in freshwater medium and it did not conserve energy to support growth from colloidal S reduction. D. acetoxidans is the first marine microorganism shown to conserve energy to support growth by coupling the complete oxidation of organic compounds to the reduction of Fe(III) or Mn(IV). Thus, D. acetoxidans provides a model enzymatic mechanism for Fe(III) or Mn(IV) oxidation of organic compounds in marine and estuarine sediments. These findings demonstrate that 16S rRNA phylogenetic analyses can suggest previously unrecognized metabolic capabilities of microorganisms. PMID- 16348889 TI - Metabolic activity and population dynamics of rhizobia introduced into unamended and bentonite-amended loamy sand. AB - Respiration measurements showed that the cumulative amount of CO(2) respired by rhizobia introduced into sterile bentonite-amended loamy sand was significantly higher than it was in unamended loamy sand. The maintenance respiration of rhizobial cells was not influenced by the presence of bentonite clay. Carbon was used more efficiently during growth in bentonite-amended than in unamended loamy sand. The presence of bentonite clay increased the growth rate of rhizobia introduced into sterile soil. Survival studies performed in nonsterile bentonite amended loamy sand showed that the use of high (10 cells per g of dry soil) rather than lower (10 to 10 cells per g of dry soil) inoculum densities increased the final survival levels of introduced rhizobia. In unamended loamy sand, the application of 10 or 10 cells per g of dry soil resulted in similar final survival levels. Pore shape and the continuity of the water-filled pore system were suggested to largely determine the colonization rate of protective microhabitats. PMID- 16348890 TI - Regulation and Characterization of Xylanolytic Enzymes of Thermoanaerobacterium saccharolyticum B6A-RI. AB - During growth on xylan and xylose Thermoanaerobacterium saccharolyticum B6A-RI produced endoxylanase, beta-xylosidase, arabinofuranosidase, and acetyl esterase, and the first three activities appeared to be produced coordinately. During nonlimiting growth on xylan, these enzyme activities were predominantly cell associated; however, during growth on limiting concentrations of xylan, the majority of endoxylanase activity was extracellular rather than cell associated. Endoxylanase, beta-xylosidase, and arabinofuranosidase activities were induced by xylan, xylose, and arabinose, respectively. Acetyl esterase activity was constitutive, and endoxylanase activity was catabolite repressed by glucose. Extracellular endoxylanase existed as a high-molecular-weight complex (molecular weight, more than 10). When analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms, the crude endoxylanase complex was composed of at least six activity bands. Endoxylanase was purified by gel filtration with Sephacryl S-300 and affinity chromatography with xylan coupled to Sepharose CL-4B preequilibrated to 45 degrees C with 50 mM sodium acetate buffer (pH 4.0) and eluted with 0.1% soluble xylan. A single area of endoxylanase activity was identified on the zymogram; when this activity was analyzed by SDS PAGE, it was composed of a major protein with a molecular weight of approximately 160,000 and a minor protein with a molecular weight of approximately 130,000. The endoxylanase activity stained with Schiff's reagent, indicative of glycoproteins, displayed a specific activity of 41 U/mg of protein on xylan, and had pH and temperature optima of 6.0 and 70 degrees C, respectively. PMID- 16348891 TI - Conjugal gene transfer to aquatic bacteria detected by the generation of a new phenotype. AB - An experimental approach based on the assembly of genes of a catabolic pathway was used to detect transconjugants in aquatic communities. Resistance to phenylmercury acetate was established in transconjugants when wide-host-range conjugal plasmids containing merB, the gene encoding organomercurial lyase, were transferred to strains from aquatic communities that had been acclimated to inorganic mercury and thus enriched for populations containing merA, the gene encoding mercuric reductase (T. Barkay, Appl. Environ. Microbiol. 53:2725-2732, 1987). Conjugation was confirmed by using the plasmids' encoded antibiotic resistance patterns and by hybridization with a eukaryotic gene. Three merB conjugal plasmids, belonging to incompatibility groups W (pGTE16), P1 (pGTE26), and N (pGTE25), were prepared. Transfers by filter matings of pGTE16 and pGTE26 from Pseudomonas aeruginosa PA01 to indigenous strains were at efficiencies of 4.5 x 10 and 4.8 x 10 transconjugant per potential recipient, respectively. These efficiencies were from 1 to 2 orders of magnitude below those observed for intraspecies matings with genetically marked recipients. The third plasmid, pGTE25, was not stably maintained in P. aeruginosa donors, and its transfer from Escherichia coli donors was below the level of detection. Characterized transconjugant strains were shown to be Pseudomonas spp. Potential applications of the described experimental approach in the creation of bacterial populations with new catabolic capabilities in hazardous waste sites and in the detection of transfer of recombinant DNA from engineered microorganisms to indigenous bacteria are discussed. PMID- 16348892 TI - Involvement of an Intracellular Oligogalacturonate Hydrolase in Metabolism of Pectin by Clostridium thermosaccharolyticum. AB - The enzymes pectin methylesterase and polygalacturonate hydrolase, which are responsible for the initial steps of pectin degradation by Clostridium thermosaccharolyticum, were shown to be induced on the polymeric substrates pectin and pectate, as well as on oligogalacturonates, and to be repressed in the presence of glucose. The digalacturonate and trigalacturonate produced by the extracellular pectin methylesterase-polygalacturonate hydrolase complex were transported across the cytoplasmic membrane and hydrolyzed by an inducible oligogalacturonate hydrolase to galacturonate. The oligogalacturonate hydrolase was separated from the polygalacturonate hydrolase and characterized. Its temperature optimum was 65 degrees C, and its pH optimum was 6. The native molecular size was 90 kDa, and the enzyme was stable for more than 1 h at 65 degrees C. The maximum reaction rate on oligomers decreased with the increasing degree of polymerization. Galacturonate was released by hydrolysis from the nonreducing end of the oligomer. The amounts of pectinolytic enzymes produced were all strictly correlated to the amount of biomass formed. Galacturonate was metabolized via a modified Entner-Doudoroff route. PMID- 16348893 TI - Plasmid Transfer between Marine Bacteria in the Aqueous Phase and Biofilms in Reactor Microcosms. AB - Plasmid transfer of broad-host-range plasmid RP1 from marine Vibrio sp. strain S14 to marine strain SW5 under optimum conditions on the surface of nutrient plates was improved 2 orders of magnitude by using the plasmid transfer process to select an SW5 recipient more efficient than the wild type in receiving and/or maintaining the plasmid. This recipient strain, SW5H, was used to form biofilms under flow conditions on the surfaces of glass beads in reactors. The S142(RP1) donor strain was introduced to the reactors after either 48 or 170 h of biofilm formation, and production of transconjugants in the aqueous phases and biofilms without selection pressure was assessed. Plasmid transfer to the recipient cells in the biofilm was detected for biofilms formed for 170 h but not in those formed for 48 h. The plasmid transfer frequency was significantly higher (P < 0.05) among cells attached to the bead surfaces in the biofilm than among cells in the aqueous phase. PMID- 16348894 TI - Characterization of Xanthomonas campestris Pathovars by rRNA Gene Restriction Patterns. AB - Genomic DNA of 191 strains of the family Pseudomonadaceae, including 187 strains of the genus Xanthomonas, was cleaved by EcoRI endonuclease. After hybridization of Southern transfer blots with 2-acetylamino-fluorene-labelled Escherichia coli 16+23S rRNA probe, 27 different patterns were obtained. The strains are clearly distinguishable at the genus, species, and pathovar levels. The variability of the rRNA gene restriction patterns was determined for four pathovars of Xanthomonas campestris species. The 16 strains of X. campestris pv. begoniae analyzed gave only one pattern. The variability of rRNA gene restriction patterns of X. campestris pv. manihotis strains could be related to ecotypes. In contrast, the variability of patterns observed for X. campestris pv. malvacearum was not correlated with pathogenicity or with the geographical origins of the strains. The highest degree of variability of DNA fingerprints was observed within X. campestris pv. dieffenbachiae, which is pathogenic to several hosts of the Araceae family. In this case, variability was related to both host plant and pathogenicity. PMID- 16348895 TI - Enhanced Carotenoid Biosynthesis by Oxidative Stress in Acetate-Induced Cyst Cells of a Green Unicellular Alga, Haematococcus pluvialis. AB - In a green alga, Haematococcus pluvialis, a morphological change of vegetative cells into cyst cells was rapidly induced by the addition of acetate or acetate plus Fe to the vegetative growth phase. Accompanied by cyst formation, algal astaxanthin formation was more enhanced by the addition of acetate plus Fe than by the addition of acetate alone. Encystment and enhanced carotenoid biosynthesis were inhibited by either actinomycin D or cycloheximide. However, after cyst formation was induced by the addition of acetate alone, carotenoid formation could be enhanced with the subsequent addition of Fe even in the presence of the inhibitors. The Fe -enhanced carotenogenesis was inhibited by potassium iodide, a scavenger for hydroxyl radical, suggesting that hydroxyl radical formed by an iron-catalyzed Fenton reaction may be required for enhanced carotenoid biosynthesis. Moreover, it was demonstrated that four active oxygen species, singlet oxygen, superoxide anion radical, hydrogen peroxide, and peroxy radical, were capable of replacing Fe in its role in the enhanced carotenoid formation in the acetate-induced cyst. From these results, it was concluded that oxidative stress is involved in the posttranslational activation of carotenoid biosynthesis in acetate-induced cyst cells. PMID- 16348896 TI - Viability and isolation of marine bacteria by dilution culture: theory, procedures, and initial results. AB - Dilution culture, a method for growing the typical small bacteria from natural aquatic assemblages, has been developed. Each of 11 experimental trials of the technique was successful. Populations are measured, diluted to a small and known number of cells, inoculated into unamended sterilized seawater, and examined three times for the presence of 10 or more cells per ml over a 9-week interval. Mean viability for assemblage members is obtained from the frequency of growth, and many of the cultures produced are pure. Statistical formulations for determining viability and the frequency of pure culture production are derived. Formulations for associated errors are derived as well. Computer simulations of experiments agreed with computed values within the expected error, which verified the formulations. These led to strategies for optimizing viability determinations and pure culture production. Viabilities were usually between 2 and 60% and decreased with >5 mg of amino acids per liter as carbon. In view of difficulties in growing marine oligobacteria, these high values are noteworthy. Significant differences in population characteristics during growth, observed by high resolution flow cytometry, suggested substantial population diversity. Growth of total populations as well as of cytometry-resolved subpopulations sometimes were truncated at levels of near 10 cells per ml, showing that viable cells could escape detection. Viability is therefore defined as the ability to grow to that population; true viabilities could be even higher. Doubling times, based on whole populations as well as individual subpopulations, were in the 1-day to 1-week range. Data were examined for changes in viability with dilution suggesting cell cell interactions, but none could be confirmed. The frequency of pure culture production can be adjusted by inoculum size if the viability is known. These apparently pure cultures produced retained the size and apparent DNA-content characteristic of the bulk of the organisms in the parent seawater. Three cultures are now available, two of which have been carried for 3 years. The method is thus seen as a useful step for improving our understanding of typical aquatic organisms. PMID- 16348897 TI - Differential responses of nitrogen-fixing cyanobacteria to salinity and osmotic stresses. AB - Two nitrogen-fixing Anabaena strains were found to be differentially tolerant to salinity and osmotic stresses. Anabaena torulosa, a brackish-water, salt-tolerant strain, was relatively osmosensitive. Anabaena sp. strain L-31, a freshwater, salt-sensitive strain, on the other hand, displayed significant osmotolerance. Salinity and osmotic stresses affected nitrogenase activity differently. Nitrogen fixation in both of the strains was severely inhibited by the ionic, but not by the osmotic, component of salinity stress. Such differential sensitivity of diazotrophy to salinity-osmotic stresses was observed irrespective of the inherent tolerance of the two strains to salt-osmotic stress. Exogenously added ammonium conferred significant protection against salinity stress but was ineffective against osmotic stress. Salinity and osmotic stresses also affected stress-induced gene expression differently. Synthesis of several proteins was repressed by salinity stress but not by equivalent or higher osmotic stress. Salinity and osmotic stresses induced many common proteins. In addition, unique salt stress- or osmotic stress-specific proteins were also induced in both strains, indicating differential regulation of protein synthesis by the two stresses. These data show that cyanobacterial sensitivity and responses to salinity and osmotic stresses are distinct, independent phenomena. PMID- 16348898 TI - Flow cytometric analysis of marine bacteria with hoechst 33342. AB - We investigated the accuracy and precision of flow cytometric (FCM) estimates of bacterial abundances using 4', 6-diamidino-2-phenylindole (DAPI) and Hoechst 33342 (HO342, a bisbenzamide derivative) on paraformaldehyde-fixed seawater samples collected from two stations near Oahu, Hawaii. The accuracy of FCM estimates was assessed against direct counts by using epifluorescence microscopy. DAPI and HO342 differ in two aspects of their chemistry that make HO342 better suited for staining marine heterotrophic bacteria for FCM analysis. These differences are most important in studies of open-ocean ecosystems that require dual-beam FCM analysis to clearly separate heterotrophic bacterial populations from populations of photosynthetic Prochlorococcus spp. Bacterial populations were easier to distinguish from background fluorescence when stained with HO342 than when stained with DAPI, because HO342 has a higher relative fluorescence quantum yield. A substantially higher coefficient of variation of blue fluorescence, which was probably due to fluorescent complexes formed by DAPI with double-stranded RNA, was observed for DAPI-stained populations. FCM estimates averaged 2.0 and 12% higher than corresponding epifluorescence microscopy direct counts for HO342 and DAPI-stained samples, respectively. A paired-sample t test between FCM estimates and direct counts found no significant difference for HO342 stained samples but a significant difference for DAPI-stained samples. Coefficients of variation of replicate FCM abundance estimates ranged from 0.63 to 2.9% (average, 1.5%) for natural bacterial concentrations of 6 x 10 to 15 x 10 cells ml. PMID- 16348899 TI - Hydroxydiether Lipid Structures in Methanosarcina spp. and Methanococcus voltae. AB - Hydroxylated diether lipids are the most abundant lipids in Methanosarcina acetivorans, Methanosarcina thermophila, and Methanosarcina barkeri MS and Fusaro, regardless of the substrate used for growth. Structural analysis of the lipid moiety freed of polar head groups revealed that the hydroxydiether lipids of all the Methanosarcina strains were hydroxylated at position 3 of sn-2 phytanyl chains. The finding that Methanosarcina strains synthesize the same hydroxydiether structure suggests that this is a taxonomic characteristic of the genus. Methanococcus voltae produced minor amounts of the 3-hydroxydiether characteristic of Methanosarcina spp. and also the 3'-hydroxydiether described previously for Methanosaeta concilii. PMID- 16348900 TI - Effects of growth pressure and temperature on Fatty Acid composition of a barotolerant deep-sea bacterium. AB - The effects of pressure and temperature on the fatty acid composition in a barotolerant deep-sea bacterium that had branched-chain fatty acids were examined. The major fatty acids of the strain at atmospheric pressure were iso C(15:0), C(16:1), iso-C(17:0), and iso-C(17:1). As the growth pressure increased, the proportion of unsaturated fatty acid increased because of an increase in the proportion of iso-C(17:1). On the other hand, as the growth temperature decreased, the proportion of unsaturated fatty acid increased because of the increase in the proportion of C(16:1) and C(18:1). PMID- 16348901 TI - Diversity within a Colony Morphotype: Implications for Ecological Research. AB - Sets of bacterial isolates with the same colony morphologies were selected from spread plates of bacteria from deep subsurface rock samples; each set had a unique morphology. API-rapid-NFT analysis revealed that isolates within a set were the same. Fatty acid methyl ester analysis of one set of isolates clustered organisms within the same species, defining variation between isolates at the biotype (subspecies) and strain levels. Metal resistances consistently tracked with colony morphology, while antibiotic resistances were less reliable. PMID- 16348902 TI - Chromosome Polymorphisms among Strains of Hansenula polymorpha (syn. Pichia angusta). AB - Contour-clamped homogeneous electrophoresis and an embedded-agarose method of sample preparation were combined to carry out an analysis of the chromosome sets of nine strains of Hansenula polymorpha (syn. Pichia angusta). Chromosomal DNA molecules could be separated into a series of bands ranging, approximately, from 650 up to 2,200 kb in size. Polymorphism of the electrophoretic pattern was demonstrated among the strains investigated in this study. Cross-hybridization between H. polymorpha and Saccharomyces cerevisiae ribosomal DNA was also observed. PMID- 16348903 TI - Effects of Physicochemical Factors and Bacterial Colony Morphotype on Association of Vibrio vulnificus with Hemocytes of Crassostrea virginica. AB - Vibrio vulnificus is a naturally occurring marine bacterium that causes invasive disease of immunocompromised humans following the consumption of raw oysters. It is a component of the natural microbiota of Gulf Coast estuaries and has been found to inhabit tissues of oysters, Crassostrea virginica (Gmelin 1791). The interaction of V. vulnificus with oyster host defenses has not been reported in detail. We examined the interaction of V. vulnificus with phagocytic oyster hemocytes as a function of time, temperature, bacterial concentration, pretreatment with hemolymph, and V. vulnificus translucent and opaque colony morphotypes. Within these experimental parameters, the results showed that the association of V. vulnificus with hemocytes increased with time, temperature, and initial V. vulnificus/hemocyte ratio. Pretreatment of V. vulnificus with serum or an increased serum concentration did not enhance V. vulnificus-hemocyte associations, a result suggesting the absence of opsonic activity. More than 50% of hemocytes bound the translucent, avirulent morphotype, whereas 10 to 20% were associated with the opaque, virulent form, a result indicating that the degree of encapsulation was related to resistance to phagocytosis, as previously described for mammalian phagocytes. Understanding these cellular interactions may, in part, explain the persistence of V. vulnificus in oyster tissues and the ecology of V. vulnificus in estuarine environments. PMID- 16348904 TI - Conjugative Plasmid Transfer between Bacteria under Simulated Marine Oligotrophic Conditions. AB - Marine Vibrio S14 strains and an Escherichia coli strain were starved in artificial seawater (NSS) with no added carbon, nitrogen, or phosphorus. The broad-host-range plasmid RP1 was transferred between the starving S14 strains and also from the E. coli donor to the S14 recipient under oligotrophic conditions, in which mixtures of donor and recipient cells were held on Nuclepore filters either floated on NSS or held such that NSS flowed through the filter. Transconjugants were obtained from S14 donors and recipients starved for at least 15 days before being mixed together for conjugation, whereas transconjugants were recovered from the E. coli donor and S14 recipient for up to 3 days of prestarvation, but not after 5 days. Transconjugants were obtained when there were as few as about 10 and 10 cells of starving S14 donors and recipients, respectively, per ml held on the filters. Starved donor and recipient mixtures incubated at 4 or 26 degrees C, as well as those allowed to mate for 2, 5, or 24 h, all yielded numbers of transconjugants which were not significantly (P > 0.05) different. PMID- 16348905 TI - Mode of Action of Lactococcin B, a Thiol-Activated Bacteriocin from Lactococcus lactis. AB - Lactococcin B (LcnB) is a small, hydrophobic, positively charged bacteriocin produced by Lactococcus lactis subsp. cremoris 9B4. Purified LcnB has a bactericidal effect on sensitive L. lactis cells by dissipating the proton motive force and causing leakage of intracellular substrates. The activity of LcnB depends on the reduced state of the Cys-24 residue. Uptake and efflux studies of different solutes suggest that LcnB forms pores in the cytoplasmic membrane of sensitive L. lactis cells in the absence of a proton motive force. At low concentrations of LcnB, efflux of those ions and amino acids which are taken up by proton motive force-driven systems was observed. However, a 150-fold higher LcnB concentration was required for efflux of glutamate, previously taken up via a unidirectional ATP-driven transport system. Strains carrying the genetic information for the immunity protein against LcnB were not affected by LcnB. The proton motive force of immune cells was not dissipated, and no leakage of intracellular substrates could be detected. PMID- 16348906 TI - Effect of Nystatin on the Metabolism of Xylitol and Xylose by Pachysolen tannophilus. AB - Ethanol production from xylitol by resting cells of Pachysolen tannophilus was increased 40-fold in the presence of nystatin, amphotericin B, and filipin, a group of antifungal agents that alter the permeability of the plasma membrane. Furthermore, these agents had little or no effect on ethanol formation from xylitol or xylose by the cell extract. During xylose metabolism, nystatin caused the intracellular xylitol to leak out into the medium at a 23-fold-faster rate but did not affect overall xylose utilization and CO(2) evolution. These observations explain the rate of xylitol utilization by cell extract being higher than that by whole cells (J. Xu and K. B. Taylor, Appl. Environ. Microbiol. 59:231-235, 1993) as well as the relative inability of P. tannophilus to utilize xylitol to support significant ethanol production and cell growth. PMID- 16348907 TI - Triacylglycerol Profiles of Tilletia controversa and Tilletia tritici. AB - The triacylglycerol (TG) profiles of teliospores of Tilletia controversa and Tilletia tritici were examined by high-performance liquid chromatography (HPLC) and gas-liquid chromatography. Boiling isopropanol was used to ensure enzyme inactivation during homogenization. The largest lipid component as determined by thin-layer chromatography was TGs. On the basis of thin-layer chromatography of crude lipid extracts, T. controversa and T. tritici do not contain a large amount of free fatty acids. TG profiles of T. controversa and T. tritici were very similar, with 18 species of TGs resolved by HPLC and gas-liquid chromatography. In both organisms, PLL (palmitic, linoleic, linoleic) was the major component, followed by LLL (trilinolein) and PLO (palmitic, linoleic, oleic). The ratio of PLO to PLL was 1:6 and 1:4 in T. tritici and T. controversa, respectively. The TGs of both organisms contain long-chain (>22 carbons) mono- and dienoic acids. Linoleic acid was the major fatty acid found in TGs from both organisms. The differences of TGs were not considered significant for differentiation purposes. PMID- 16348908 TI - Involvement of Genes on a Megaplasmid in the Acid-Tolerant Phenotype of Rhizobium leguminosarum Biovar Trifolii. AB - The acid-tolerant Rhizobium leguminosarum biovar trifolii strain ANU1173 exhibited several new phenotypes when cured of its symbiotic (Sym) plasmid and the second largest megaplasmid. Strain P22, which has lost these two plasmids, had reduced exopolysaccharide production and cell mobility on TY medium. The parent strain ANU1173 was able to grow easily in laboratory media at pH 4.5, whereas the derivative strain P22 was unable to grow in media at a pH of <4.7. The intracellular pH of strain ANU1173 was 6.8 when the external pH was 4.5. In contrast, strain P22 had an acidic intracellular pH of <6.4 when the external pH was <5.5. Strain P22 had a dramatically increased membrane permeability to protons and decreased proton extrusion activity. Analysis with sodium dodecyl sulfate-polyacrylamide gels showed that strain P22 lacked a slow-migrating lipopolysaccharide (LPS) banding group which was present in the parent strain. Mobilization of the second largest megaplasmid of strain ANU1173 back into strain P22 restored the altered LPS structure and physiological characteristics of strain P22. Mobilization of the Sym plasmid of strain ANU1173 into strain P22 showed that the second largest megaplasmid of strain ANU1173 was required for the establishment of nitrogen-fixing nodules on Trifolium repens and Trifolium subterraneum. Furthermore, an examination of a large number of specific exopolysaccharide- or LPS-deficient Rhizobium mutants did not show a positive correlation between exopolysaccharide or LPS synthesis and acid tolerance. PMID- 16348909 TI - Metabolic Pathways in Methanococcus jannaschii and Other Methanogenic Bacteria. AB - Eleven strains of methanogenic bacteria were divided into two groups on the basis of the directionality (oxidative or reductive) of their citric acid pathways. These pathways were readily identified for most methanogens from the patterns of carbon atom labeling in glutamate, following growth in the presence of [2 C]acetate. All used noncyclic pathways, but members of the family Methanosarcinaceae were the only methanogens found to use the oxidative direction. Methanococcus jannaschii failed to incorporate carbon from acetate despite transmembrane equilibration comparable to other weak acids. This organism was devoid of detectable activities of the acetate-incorporating enzymes acetyl coenzyme A synthetase, acetate kinase, and phosphotransacetylase. However, incorporation of [1-C]-, [2-C]-, or [3-C]pyruvate during the growth of M. jannaschii was possible and resulted in labeling patterns indicative of a noncyclic citric acid pathway operating in the reductive direction to synthesize amino acids. Carbohydrates were labeled consistent with glucogenesis from pyruvate. Leucine, isoleucine, phenylalanine, lysine, formate, glycerol, and mevalonate were incorporated when supplied to the growth medium. Lysine was preferentially incorporated into the lipid fraction, suggesting a role as a phytanyl chain precursor. PMID- 16348910 TI - Production of Specifically Labeled Compounds by Methanobacterium espanolae Grown on H(2)-CO(2) plus [C]Acetate. AB - Methanobacterium espanolae, an acidiphilic methanogen, required acetate for maximal growth on H(2)-CO(2). In the presence of 5 to 15 mM acetate, at a growth pH of 5.5, the mu(max) was 0.05 h. M. espanolae consumed 12.3 mM acetate during 96 h of incubation at 35 degrees C with shaking at 100 rpm. At initial acetate levels of 2.5 to 10.0 mM, the amount of biomass produced was dependent on the amount of acetate in the medium. C nuclear magnetic resonance spectra of protein hydrolysates obtained from cultures grown on [1-C]- or [2-C]acetate indicated that an incomplete tricarboxylic acid pathway, operating in the reductive direction, was functional in this methanogen. The amino acids were labeled with a very high degree of specificity and at greater than 90% enrichment levels. Less than 2% label randomization occurred between positions primarily labeled from either the carboxyl or methyl group of acetate, and very little label was transferred to positions primarily labeled from CO(2). The labeling pattern of carbohydrates was typical for glucogenesis from pyruvate. This methanogen, by virtue of the properties described above and its ability to incorporate all of the available acetate (10 mM or lower) from the growth medium, has advantages over other microorganisms for use in the production of specifically labeled compounds. PMID- 16348911 TI - Effects of pH and Osmotic Stress on Cellular Polyamine Contents in the Soybean Rhizobia Rhizobium fredii P220 and Bradyrhizobium japonicum A1017. AB - Homospermidine is a polyamine present in its highest concentrations in root nodule bacteria. By using the soybean rhizobia Rhizobium fredii P220 and Bradyrhizobium japonicum A1017, the effects of the pH and osmolarity of the medium on rhizobial growth and cellular polyamine contents were investigated. Elevation of medium pH repressed the growth of slowly growing B. japonicum A1017 and resulted in a slight increase in cellular putrescine, while homospermidine content was not significantly affected. In contrast, in fast-growing R. fredii P220, which showed good growth over a wide range of the medium pHs from 4.0 to 9.5, homospermidine content increased with the lowering of the medium pH. Under the acid-stressed conditions, cellular Mg content in strain P220 also increased. Strain P220 was able to grow in NaCl concentrations up to 0.4 M, while strain A1017 did not grow in media containing 0.15 M NaCl. Glutamic acid and K contents of salt-tolerant P220 cells increased in response to NaCl concentrations, but homospermidine and Mg contents were inversely related to the NaCl concentrations. External salinity had no effect on the contents of other polyamines in P220 cells. On the basis of osmotic strength, NaCl, KCl, sucrose, or glycerol induced similar decreases in cellular homospermidine content. These results suggested that the cellular levels of homospermidine in strain P220 may be regulated by mechanisms related to their pH and osmotic tolerance. PMID- 16348912 TI - Growth of syntrophic propionate-oxidizing bacteria with fumarate in the absence of methanogenic bacteria. AB - Oxidation of succinate to fumarate is an energetically difficult step in the biochemical pathway of propionate oxidation by syntrophic methanogenic cultures. Therefore, the effect of fumarate on propionate oxidation by two different propionate-oxidizing cultures was investigated. When the methanogens in a newly enriched propionate-oxidizing methanogenic culture were inhibited by bromoethanesulfonate, fumarate could act as an apparent terminal electron acceptor in propionate oxidation. C-nuclear magnetic resonance experiments showed that propionate was carboxylated to succinate while fumarate was partly oxidized to acetate and partly reduced to succinate. Fumarate alone was fermented to succinate and CO(2). Bacteria growing on fumarate were enriched and obtained free of methanogens. Propionate was metabolized by these bacteria when either fumarate or Methanospirillum hungatii was added. In cocultures with Syntrophobacter wolinii, such effects were not observed upon addition of fumarate. Possible slow growth of S. wolinii on fumarate could not be demonstrated because of the presence of a Desulfovibrio strain which grew rapidly on fumarate in both the absence and presence of sulfate. PMID- 16348913 TI - Metabolism of p-Cresol by the Fungus Aspergillus fumigatus. AB - The fungus Aspergillus fumigatus ATCC 28282 was shown to grow on p-cresol as its sole source of carbon and energy. A pathway for metabolism of this compound was proposed. This has protocatechuate as the ring-fission substrate with cleavage and metabolism by an ortho-fission pathway. The protocatechuate was formed by two alternative routes, either by initial attack on the methyl group, which is oxidized to carboxyl, followed by ring-hydroxylation, or by ring-hydroxylation as the first step with subsequent oxidation of 4-methylcatechol to the acid. The pathway was elucidated from several pieces of evidence. A number of compounds, including 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid, protocatechuic acid, protocatechualdehyde, and 4-methylcatechol, appeared transiently in the medium during growth on p-cresol. These compounds were oxidized without lag by p-cresol-grown cells but not by succinate-grown cells. Enzyme activities for most of the proposed steps were demonstrated in cell extracts after growth on p-cresol, and the products of these activities were identified. None of the activities were found in succinate-grown cells. PMID- 16348914 TI - Degradation of degradable starch-polyethylene plastics in a compost environment. AB - The degradation performance of 11 types of commercially produced degradable starch-polyethylene plastic compost bags was evaluated in municipal yard waste compost sites at Iowa State University (Ames) and in Carroll, Dubuque, and Grinnell, Iowa. Masterbatches for plastic production were provided by Archer Daniels Midland Co. (Decatur, Ill.), St. Lawrence Starch Co. Ltd. (Mississauga, Ontario, Canada), and Fully Compounded Plastics (Decatur, Ill.). Bags differed in starch content (5 to 9%) and prooxidant additives (transition metals and a type of unsaturated vegetable oil). Chemical and photodegradation properties of each material were evaluated. Materials from St. Lawrence Starch Co. Ltd. and Fully Compounded Plastics photodegraded faster than did materials from Archer Daniels Midland Co., whereas all materials containing transition metals demonstrated rapid thermal oxidative degradation in 70 degrees C-oven (dry) and high temperature, high-humidity (steam chamber) treatments. Each compost site was seeded with test strips (200 to 800 of each type) taped together, which were recovered periodically over an 8- to 12-month period. At each sampling date, the compost row temperature was measured (65 to 95 degrees C), the location of the recovered test strip was recorded (interior or exterior), and at least four strips were recovered for evaluation. Degradation was followed by measuring the change in polyethylene molecular weight distribution via high-temperature gel permeation chromatography. Our initial 8-month study indicated that materials recovered from the interior of the compost row demonstrated very little degradation, whereas materials recovered from the exterior degraded well. In the second-year study, however, degradation was observed in several plastic materials recovered from the interior of the compost row by month 5 at the Carroll site and almost every material by month 12 at the Grinnell site. The plastic bags collected from each community followed a similar degradation pattern. To our knowledge, this is the first scientific study demonstrating significant polyethylene degradation by these materials in a compost environment. PMID- 16348915 TI - Isolation and characterization of a mo -reducing bacterium. AB - A Mo -reducing bacterium (strain 48), which grew on medium supplemented with 200 mM Mo, was isolated from stream water obtained from Chengkau, Malaysia. The chemical properties of strain 48 conform to the characteristics of Enterobacter cloacae. Under anaerobic conditions in the glucose-yeast extract medium containing phosphate ion (2.9 mM) and Mo (10 mM), the bacterium reduced Mo to form molybdenum blue. Approximately 27% of Mo added to the medium was reduced after 28 h of cultivation. The reduction of Mo with glucose as an electron donor was strongly inhibited by iodoacetic acid, sodium fluoride, and sodium cyanide, suggesting an involvement of the glycolytic pathway and electron transport in Mo reduction. NADH and N,N,N',N' -tetramethyl-p-phenylenediamine served as electron donors for Mo reduction. When NADH was used as an electron donor, at first cytochrome b in the cell extract was reduced, and then molybdenum blue was formed. Sodium cyanide strongly inhibited Mo reduction by NADH (5 mM) but not the reduction of cytochrome b in the cell extract, suggesting that the reduced component of the electron transport system after cytochrome b serves as an electron donor for Mo reduction. Both ferric and stannous ions strongly enhanced the activity of Mo reduction by NADH. PMID- 16348916 TI - Quantitative and Qualitative Analysis of Bacteria in Er(III) Solution by Thin Film Magnetopheresis. AB - Magnetic deposition, quantitation, and identification of bacteria reacting with the paramagnetic trivalent lanthanide ion, Er, was evaluated. The magnetic deposition method was dubbed thin-film magnetopheresis. The optimization of the magnetic deposition protocol was accomplished with Escherichia coli as a model organism in 150 mM NaCl and 5 mM ErCl(3) solution. Three gram-positive bacteria, Staphylococcus epidermidis, Staphylococcus saprophyticus, and Enterococcus faecalis, and four gram-negative bacteria, E. coli, Pseudomonas aeruginosa, Proteus mirabilis, and Klebsiella pneumoniae, were subsequently investigated. Quantitative analysis consisted of the microscopic cell count and a scattered light scanning of the magnetically deposited material aided by the computer data acquisition system. Qualitative analysis consisted of Gram stain differentiation and fluorescein isothiocyanate staining in combination with selected antisera against specific types of bacteria on the solid substrate. The magnetic deposition protocol allowed quantitative detection of E. coli down to the concentration of 10 CFU ml, significant in clinical diagnosis applications such as urinary tract infections. Er did not interfere with the typical appearance of the Gram-stained bacteria nor with the antigen recognition by the antibody in the immunohistological evaluations. Indirect antiserum-fluorescein isothiocyanate labelling correctly revealed the presence of E. faecalis and P. aeruginosa in the magnetically deposited material obtained from the mixture of these two bacterial species. On average, the reaction of gram-positive organisms was significantly stronger to the magnetic field in the presence of Er than the reaction of gram negative organisms. The thin-film magnetophoresis offers promise as a rapid method for quantitative and qualitative analysis of bacteria in solutions such as urine or environmental water. PMID- 16348917 TI - Bacterial Florae in Larvae of the Lake Fly Chironomus plumosus. AB - Chironomus plumosus midge larvae were collected from Lake Winnebago, Wisconsin, for 10 weeks in the summer of 1985 and 10 weeks in the summer of 1986 in order to determine their bacterial floras. Altogether, 18 genera and 29 species of bacteria were found and identified. Some spirillum-like bacteria which were not isolated in culture were found by electron microscopy. Scanning electron micrographs of sectioned larvae revealed bacteria throughout the intestines. Gram positive organisms were most prevalent in the larvae in the early part of the summer, and gram-negative organisms were most prevalent during the later part. Larvae accumulate bacteria within their intestines. PMID- 16348918 TI - Visual reading method for detection of bacterial tannase. AB - Tannase activity of bacteria capable of degrading tannin-protein complexes was determined by a newly developed visual reading method. The method is based on two phenomena: (i) the ability of tannase to hydrolyze methyl gallate to release free gallic acid and (ii) the green to brown coloration of gallic acid after prolonged exposure to oxygen in an alkaline condition. The method has been successfully used to detect the presence of tannase in the cultures of bacteria capable of degrading tannin-protein complexes. PMID- 16348919 TI - A new way of producing isomalto-oligosaccharide syrup by using the transglycosylation reaction of neopullulanase. AB - A new way of producing isomalto-oligosaccharide syrup from starch was developed. Isomalto-oligosaccharides contain one or more alpha-(1-->6)-glucosidic linkages with or without alpha-(1-->4)-glucosidic linkages. The isomalto-oligosaccharide syrups are receiving increased attention as food additives because it is thought that they help prevent dental caries and improve human intestinal microflora, acting as a growth factor for bifidobacteria. The new system for production of isomalto-oligosaccharide syrup is based on the strong alpha-(1-->6) transglycosylation reaction of neopullulanase. Bacillus subtilis saccharifying alpha-amylase was simultaneously used with neopullulanase to improve the yield of isomalto-oligosaccharides. The yield of isomalto-oligosaccharides was increased to more than 60%, compared with a yield of 45.0% obtained by the conventional system. To reduce the costs, the use of immobilized neopullulanase was investigated. Almost the same yield of isomalto-oligosaccharides was obtained by using immobilized neopullulanase. PMID- 16348920 TI - Soluble Methane Monooxygenase Production and Trichloroethylene Degradation by a Type I Methanotroph, Methylomonas methanica 68-1. AB - A methanotroph (strain 68-1), originally isolated from a trichloroethylene (TCE) contaminated aquifer, was identified as the type I methanotroph Methylomonas methanica on the basis of intracytoplasmic membrane ultrastructure, phospholipid fatty acid profile, and 16S rRNA signature probe hybridization. Strain 68-1 was found to oxidize naphthalene and TCE via a soluble methane monooxygenase (sMMO) and thus becomes the first type I methanotroph known to be able to produce this enzyme. The specific whole-cell sMMO activity of 68-1, as measured by the naphthalene oxidation assay and by TCE biodegradation, was comparatively higher than sMMO activity levels in Methylosinus trichosporium OB3b grown in the same copper-free conditions. The maximal naphthalene oxidation rates of Methylomonas methanica 68-1 and Methylosinus trichosporium OB3b were 551 +/- 27 and 321 +/- 16 nmol h mg of protein , respectively. The maximal TCE degradation rates of Methylomonas methanica 68-1 and Methylosinus trichosporium OB3b were 2,325 +/- 260 and 995 +/- 160 nmol h mg of protein, respectively. The substrate affinity of 68-1 sMMO to naphthalene (K(m), 70 +/- 4 muM) and TCE (K(m), 225 +/- 13 muM), however, was comparatively lower than that of the sMMO of OB3b, which had affinities of 40 +/- 3 and 126 +/- 8 muM, respectively. Genomic DNA slot and Southern blot analyses with an sMMO gene probe from Methylosinus trichosporium OB3b showed that the sMMO genes of 68-1 have little genetic homology to those of OB3b. This result may indicate the evolutionary diversification of the sMMOs. PMID- 16348921 TI - Seasonal Dynamics of Bacterial Colonization of Cotton Fiber and Effects of Moisture on Growth of Bacteria within the Cotton Boll. AB - A highly replicated 3-year field study was conducted to determine the seasonal patterns of bacterial colonization of cotton fiber from the time of dehiscence of the bolls (the point at which the bolls just begin to open) through harvest and commercial ginning. Bacterial numbers on fiber samples from 16 plots were determined by dilution pour plating with tryptic soy agar containing cycloheximide, and numbers of gram-negative bacteria were determined by plating on tryptic soy agar containing vancomycin and cycloheximide. Populations of bacteria varied from year to year, but in all three seasons the pattern of colonization was generally a pattern consisting of a rapid increase following opening of the bolls and a more or less stable number thereafter throughout the growing season. Gram-negative bacteria accounted for 50% or more of the recoverable bacterial population. We hypothesized that the luxuriant bacterial flora developed as a result of the availability of sufficient free water in the bolls to allow bacterial proliferation with the carbon sources remaining after fiber maturation. Therefore, laboratory experiments were conducted to determine the threshold moisture level allowing growth of bacteria on fiber in the bolls. Bacterial proliferation occurred when as little as 2% moisture was added to air dried fiber. Using simulated bolls, we demonstrated bacterial growth resulting from dew formation on fiber held in controlled-humidity chambers. PMID- 16348922 TI - Isolation and diversity of actinomycetes in the chesapeake bay. AB - Chesapeake Bay was investigated as a source of actinomycetes to screen for production of novel bioactive compounds. The presence of relatively large populations of actinoplanetes (chemotype II/D actinomycetes) in Chesapeake Bay sediment samples indicates that it is an eminently suitable ecosystem from which to isolate actinomycetes for screening programs. Actinomycetes were isolated from sediment samples collected in Chesapeake Bay with an isolation medium containing nalidixic acid, which proved to be more effective than heat pretreatment of samples. Actinomycete counts ranged from a high of 1.4 x 10 to a low of 1.8 x 10 CFU/ml of sediment. Actinomycetes constituted 0.15 to 8.63% of the culturable microbial community. The majority of isolates from the eight stations studied were actinoplanetes (i.e., chemotype II/D), and 249 of these isolates were obtained in a total of 298 actinomycete isolates. Antimicrobial activity profiles indicated that diverse populations of actinoplanetes were present at each station. DNA hybridization studies showed considerable diversity among isolates between stations, but indicated that actinoplanete strains making up populations at nearby stations were more similar to each other than to populations sampled at distant stations. The diversity of actinoplanetes and the ease with which these organisms were isolated from Chesapeake Bay sediments make this a useful source of these actinomycetes. PMID- 16348923 TI - Binding of extracellular carboxymethylcellulase activity from the marine shipworm bacterium to insoluble cellulosic substrates. AB - The binding of extracellular endoglucanase, a carboxymethylcellulase (CMCase), produced by the marine shipworm bacterium to insoluble cellulose substrates was investigated. Up to 70% of CMCase activity bound to cellulosic substrates, and less than 10% bound to noncellulosic substrates. CMCase binding to cellulose was enhanced in basal salt medium or sodium phosphate buffer containing 0.5 M NaCl. Increased cellulose particle size correlated with decreased CMCase binding. Also, cellulose treated with either 5 N NaOH or commercial cellulase reduced the CMCase binding to these surfaces. Pretreatment of CMCase preparations with 0.01% sodium dodecyl sulfate, 5% beta-mercaptoethanol, and 5 mM EDTA or ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) had little effect on binding to cellulose. While pretreatment of CMCase with trypsin, chymotrypsin, and pronase had little effect on CMCase enzymatic activity, the ability to bind to cellulose was greatly diminished by these treatments. PMID- 16348924 TI - Sorption of heavy metals to the filamentous bacterium thiothrix strain A1. AB - A study was undertaken to determine the ability of the filamentous bacterium Thiothrix strain A1 to sorb heavy metals from solution. Cells of Thiothrix strain A1 were harvested, washed, and suspended in solutions of metals. After an equilibration period, biomass was separated from solution and the metal content in acid-digested cells and/or filtrates was determined by atomic absorption spectrophotometry. Sorption of nickel and zinc was very rapid; most of the sorbed metal was bound in less than 10 min. The sorption data for copper fit the Freundlich isotherm, and nickel and zinc data fit biphasic Freundlich isotherms. Sorption of both nickel and zinc was dependent on cell age. Cells harvested 24 h after inoculation sorbed approximately one-half of the amount of metal per gram cell protein than did cells harvested after 48, 72, or 96 h. Calcium and magnesium effectively competed with zinc for binding sites, whereas potassium had only a slight effect on the capacity of cells to sorb zinc. The primary mechanism of metal sorption apparently was ion exchange, because 66 to 75% of nickel or zinc could be desorbed by placing metal-laden cells in a solution of 5 mM CaCl(2). A competition experiment with nickel and zinc indicated that both metals occupied the same sorption sites. The strong chelating agents EDTA and NTA effectively prevented metal uptake, but lactate enhanced the uptake of nickel. Thiothrix strain A1 grown in nickel-containing medium had a relatively low uptake of nickel compared with uptake by resting cells suspended in a simple buffer solution. PMID- 16348925 TI - Examination of Lipopolysaccharide (O-Antigen) Populations of Thiobacillus ferrooxidans from Two Mine Tailings. AB - Net acid-generating capacities of 39.74 kg of H(2)SO(4) per ton (ca. 0.05 kg/kg) (pH 2.68) for the Lemoine copper mine tailings (closed ca. 8 years ago; located 40 km west of Chibougamau, Quebec, Canada) and 16.07 kg of H(2)SO(4) per ton (ca. 0.02 kg/kg) (pH 3.01) for the Copper Rand tailings (in current use and 50 km distant [east] from those of Lemoine) demonstrate that these sulfide tailings can support populations of acidophilic thiobacilli. Oxidized regions in both tailings environments were readily visible, were extremely acidic (Lemoine, pH 2.36; Copper Rand, pH 3.07), and provided natural isolates for our study. A 10% (wt/vol) oxalic acid treatment, which solubilizes both ferric sulfate and ferric hydroxide precipitates (B. Ramsay, J. Ramsay, M. deTremblay, and C. Chavarie, Geomicrobiol. J. 6:171-177, 1988), enabled the recovery of intact bacterial cells from the tailings material and from liquid synthetic medium for lipopolysaccharide analysis. No viable cells could be cultured after this oxalic acid treatment. Sodium dodecyl sulfate-polyacrylamide gel electro-phoretic profiles of lipopolysaccharides extracted from the Lemoine tailings were complex, indicating a heterogeneous population of Thiobacillus ferrooxidans. Six T. ferrooxidans subspecies as identified by lipopolysaccharide analysis (i.e., lipopolysaccharide chemotypes) were eventually isolated from a total of 112 cultures from the Lemoine tailings. Using the same isolate and lipopolysaccharide typing techniques, we identified only a single lipopolysaccharide chemotype from 20 cultures of T. ferrooxidans isolated from the Copper Rand tailings. This homogeneity of lipopolysaccharide chemotype was much different from what was found for the older Lemoine tailings and may reflect a progressive lipopolysaccharide heterogeneity of Thiobacillus isolates as tailings leach and age. PMID- 16348926 TI - Comparison of Nucleic Acid Hybridization and Fluorometry for Measurement of the Relationship between RNA/DNA Ratio and Growth Rate in a Marine Bacterium. AB - Continuous culture of Pseudomonas stutzeri Zobell, a marine denitrifying bacterium, was used to determine the relationship between growth rate and nucleic acid content. The trend of decreasing RNA content with decreasing growth rate, well known for enteric organisms, was found to occur in P. stutzeri Zobell as well, even at very long generation times such as those thought to occur in the oligotrophic ocean. When assayed by ethidium bromide fluorescence, the total RNA/DNA ratio was linear for generation times between 6 and 60 h. We also developed a 200-bp nucleic acid probe (with species-specific potential) for a portion of the 23S rRNA gene of P. stutzeri Zobell, which was used to quantify rRNA and rDNA by hybridization in the same continuous cultures. The rRNA/rDNA ratio also exhibited a decrease with decreasing growth rate, but the relationship, although significant, was not simply linear. The sensitivity and accuracy of the two methods are compared, and the potential for species specificity in future hybridizations is discussed. PMID- 16348927 TI - Correlative Association between Resident Plasmids and the Host Chromosome in a Diverse Agrobacterium Soil Population. AB - Soil samples collected from a fallow field which had not been cultivated for 5 years harbored a population of Agrobacterium spp. estimated at 3 x 10 CFU/g. Characterization of 72 strains selected from four different isolation media showed the presence of biovar 1 (56%) and bv. 2 (44%) strains. Pathogenicity assays on five different test plants revealed a high proportion (33%) of tumorigenic strains in the resident population. All tumorigenic strains belonged to bv. 1. Differentiation of the strains by restriction fragment length polymorphism analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of cellular proteins, and utilization patterns of 95 carbon substrates (Biolog GN microplate) revealed a diversified bv. 1 population, composed of five distinct chromosomal backgrounds (chr A, C, D, E, and F), and a homogeneous bv. 2 population (chr B). chr A, B, C, and D were detected at similar levels throughout the study site. According to opine metabolism, pathogenicity, and agrocin sensitivity, chr A strains carried a nopaline Ti plasmid (pTi), whereas chr C strains had an octopine pTi. In addition, four of six nontumorigenic bv. 1 strains (two chr D, one chr E, and one chr F) had distinct and unusual opine catabolism patterns. chr B (bv. 2) strains were nonpathogenic and catabolized nopaline. Although agrocin sensitivity is a pTi-borne trait, 14 chr B strains were sensitive to agrocin 84, apparently harboring a defective nopaline pTi similar to pAtK84b. The other two chr B strains were agrocin resistant. The present analysis of chromosomal and plasmid phenotypes suggests that in this Agrobacterium soil population, there is a preferential association between the resident plasmids and their bacterial host. PMID- 16348928 TI - H(2)-CO(2)-Dependent Anaerobic O-Demethylation Activity in Subsurface Sediments and by an Isolated Bacterium. AB - The ability of microorganisms in sediments from the Atlantic Coastal Plain to biodegrade methoxylated aromatic compounds was examined. O-demethylation activity was detected in deep (121- and 406-m) sediments, as well as in the surface soil. A syringate-demethylating consortium, containing at least three types of bacteria, was enriched from a deep-sediment sample in a medium containing syringate as the sole organic carbon source and with a N(2)-CO(2) atmosphere. An isolate which demethylated syringate was obtained from the enrichment on an agar medium incubated under a H(2)-CO(2) but not a N(2)-CO(2) or N(2) atmosphere. O demethylation of syringate of this isolate was dependent on the presence of both H(2) and CO(2) in the gas phase. The metabolism of syringate occurred in a sequential manner: methylgallate accumulated transiently before it was converted to gallate. Mass balance analysis suggests that the stoichiometry of the reaction in this isolate proceeds in accordance with the following generalized equation: C(7)H(3)O(3)(OCH(3))(n) + nHCO(3) + nH(2) --> C(7)H(3)O(3)(OH)(n) + nCH(3)COO + nH(2)O. PMID- 16348929 TI - Resistance to Toxins from Bacillus thuringiensis subsp. kurstaki Causes Minimal Cross-Resistance to B. thuringiensis subsp. aizawai in the Diamondback Moth (Lepidoptera: Plutellidae). AB - Repeated exposure in the field followed by laboratory selection produced 1,800- to >6,800-fold resistance to formulations of Bacillus thuringiensis subsp. kurstaki in larvae of the diamondback moth, Plutella xylostella. Four toxins from B. thuringiensis subsp. kurstaki [CryIA(a), CryIA(b), CryIA(c), and CryIIA] caused significantly less mortality in resistant larvae than in susceptible larvae. Resistance to B. thuringiensis subsp. kurstaki formulations and toxins did not affect the response to CryIC toxin from B. thuringiensis subsp. aizawai. Larvae resistant to B. thuringiensis subsp. kurstaki showed threefold cross resistance to formulations of B. thuringiensis subsp. aizawai containing CryIC and CryIA toxins. This minimal cross-resistance may be caused by resistance to CryIA toxins shared by B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. aizawai. PMID- 16348930 TI - Novel Allylic Oxidation of alpha-Cedrene to sec-Cedrenol by a Rhodococcus Strain. AB - A bacterial strain, designated KSM-7358, that can use alpha-cedrene for growth was isolated. The strain was identified as a member of the genus Rhodococcus and catalyzed the novel allylic oxidation of alpha-cedrene regiospecifically to produce (R)-10-hydroxycedrene (sec-cedrenol) with a very high yield. alpha Curcumene was also produced as a possible metabolite of sec-cedrenol. A possible pathway for the microbial conversion of alpha-cedrene to sec-cedrenol and alpha curcumene is proposed. PMID- 16348931 TI - Growth, nitrogen fixation, and spectral attenuation in cultivated trichodesmium species. AB - Physiological studies of Trichodesmium species have been hindered by difficulties in maintaining actively growing, nitrogen-fixing cultures. Previous cultivation successes have not been widely duplicated. We present here a simple modified seawater medium and handling techniques which have been used to maintain actively growing Trichodesmium thiebautii in laboratory culture for over 1 year. The cultured population, isolated from coastal Atlantic waters, has a growth rate of 0.23 division day and exhibits light-dependent nitrogen fixation during exponential growth. Various morphologies, including solitary trichomes, and aggregates (spherical puffs and fusiform tufts) are present during growth. Spectral and scalar irradiance were measured within naturally occurring (coastal Atlantic) aggregates with small (diameter, 50 to 70 mum) spherical fiber-optic sensors. In contrast to naturally occurring puffs, cultivated Trichodesmium aggregates exhibited spectral properties consistent with low-light adaptation. Cultivated puff-type aggregates were also examined by using oxygen microelectrodes. The simple medium and approach used for cultivation should be easily reproducible and amenable to further manipulations and modifications useful for physiological studies of Trichodesmium spp. in culture. PMID- 16348932 TI - Purification and Characterization of Alkaline Xylanases from Bacillus polymyxa. AB - By applying different classical and fast protein liquid chromatographic techniques, three xylanases (beta-1,4-d-xylan xylanhydrolase) were purified to homogeneity from the extracellular enzymatic complex of Bacillus polymyxa. The three enzymes (X(34)C, X(34)E, and X(22)) were small proteins of 34, 34, and 22 kDa and basic pIs 9.3, >9.3, and 9.0, respectively. X(34)C and X(34)E are closely related and seem to be isoforms of the same enzyme. However, they differ in some characteristics. The three enzymes had different pH and temperature optima. One of them, X(34)E, showed a high thermal stability. The V(max) values determined for X(34)C, X(34)E, and X(22) enzymes on oat spelts xylan were 14.9, 85.5, and 64.0 U mg, respectively, and 16.1, 62.0, and 150.6 U mg on birchwood xylan. When oat spelts xylan was the substrate used, K(m) values of 3.4, 2.4, and 1.9 mg ml were obtained for X(34)C, X(34)E, and X(22) enzymes, respectively, and 0.65, 6.3, and 0.32 mg ml were the respective K(m) values determined with birchwood xylan as the substrate. The enzymes were nondebranching endo-beta-xylanases. Xylose was one of the products of xylan hydrolysis by xylanases X(34)C and X(34)E, but this monosaccharide was not released by X(22) enzyme. However, neither of the enzymes was able to degrade xylobiose. PMID- 16348933 TI - Plantaricins S and T, Two New Bacteriocins Produced by Lactobacillus plantarum LPCO10 Isolated from a Green Olive Fermentation. AB - Twenty-six strains of Lactobacillus plantarum isolated from green olive fermentations were tested for cross-antagonistic activities in an agar drop diffusion test. Cell-free supernatants from four of these strains were shown to inhibit the growth of at least one of the L. plantarum indicator strains. L. plantarum LPCO10 provided the broadest spectrum of activity and was selected for further studies. The inhibitory compound from this strain was active against some gram-positive bacteria, including clostridia and propionibacteria as well as natural competitors of L. plantarum in olive fermentation brines. In contrast, no activity against gram-negative bacteria was detected. Inhibition due to the effect of organic acids, hydrogen peroxide, or bacteriophages was excluded. Since the inhibitory activity of the active supernatant was lost after treatment with various proteolytic enzymes, this substance could be classified as a bacteriocin, designated plantaricin S. Plantaricin S was also sensitive to glycolytic and lipolytic enzymes, suggesting that it was a glycolipoprotein. It exhibited a bactericidal and nonbacteriolytic mode of action against indicator cells. This bacteriocin was heat stable (60 min at 100 degrees C), active in a pH range of 3.0 to 7.0, and also stable in crude culture supernatants during storage. Ultrafiltration studies indicated that plantaricin S occurred as multimolecular aggregates and that the size of the smallest active form is between 3 and 10 kDa. In sodium dodecyl sulfate-polyacrylamide gels, plantaricin S migrated as a peptide of ca. 2.5 kDa. Maximum production of plantaricin S was obtained in a fermentor system in unregulated pH and log-phase cultures of L. plantarum LPCO10 in MRS broth plus 4% NaCl. In these culture conditions, a second bacteriocin (designated plantaricin T) was produced in late-stationary-phase cultures of L. plantarum LPCO10. On the basis of its biological activity, its sensitivity to various enzymes, and its molecular weight (lower than that of plantaricin S) as assessed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, plantaricin T appeared different from plantaricin S. Curing experiments with L. plantarum LPCO10 resulted in the appearance of variants that no longer produced either of the two bacteriocins but that were still immune to both of them. PMID- 16348934 TI - Complete oxidation of propionate, valerate, succinate, and other organic compounds by newly isolated types of marine, anaerobic, mesophilic, gram negative, sulfur-reducing eubacteria. AB - Anaerobic enrichment cultures with either propionate, succinate, lactate, or valerate and elemental sulfur and inocula from shallow marine or deep-sea sediments were dominated by rod-shaped motile bacteria after three transfers. By application of deep-agar dilutions, five eubacterial strains were obtained in pure culture and designated Kyprop, Gyprop, Kysw2, Gylac, and Kyval. All strains were gram negative and grew by complete oxidation of the electron donors and concomitant stoichiometric reduction of elemental sulfur to hydrogen sulfide. The isolates used acetate, propionate, succinate, lactate, pyruvate, oxaloacetate, maleate, glutamate, alanine, aspartate, and yeast extract. All isolates, except strain Gylac, used citrate as an electron donor but valerate was oxidized only by strain Kyval. Fumarate and malate were degraded by all strains without an additional electron donor or acceptor. Kyprop, Gyprop, and Gylac utilized elemental sulfur as the sole inorganic electron acceptor, while Kysw2 and Kyval also utilized nitrate, dimethyl sulfoxide, or Fe(III)-citrate as an electron acceptor. PMID- 16348935 TI - Identification of the sources of energy for nitrogen fixation and physiological characterization of nitrogen-fixing members of a marine microbial mat community. AB - Experimental manipulations of a microbial mat community were performed to determine sources of energy and reductant used for nitrogen fixation and to physiologically characterize the responsible diazotrophs. The dominant photolithotrophic members of this community were nonheterocystous cyanobacteria, but other potential nitrogen-fixing microorganisms were also present. Pronounced diel variability in rates of acetylene reduction was observed, with nighttime rates a factor of three to four higher than daytime rates. Acetylene reduction measured at night was dependent upon the occurrence of oxygenic photosynthesis the preceding day; mats incubated in the dark during the daytime reduced acetylene at rates comparable to those of light-incubated mats but were not able to reduce acetylene at the normally high rates the following night. The addition of various exogenous carbon compounds to these dark-incubated mats did not elicit nighttime acetylene reduction. Nighttime acetylene reduction apparently proceeds under anoxic conditions in these mats; the highest rates of acetylene reduction occur late at night. Additions of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (an inhibitor of oxygenic photosynthesis) to mats resulted in a pronounced stimulation of acetylene reduction during the day, but acetylene reduction the next night proceeded at greatly reduced rates (relative to untreated mats). This daytime stimulation, under the 3-(3,4-dichlorophenyl)-1,1-dimethylurea-induced anoxic conditions in the experimentally treated mats, was light dependent. These results suggest that nitrogen fixation in these mats may be attributed to the activities of nonheterocystous cyanobacteria utilizing storage products of oxygenic photosynthesis under anoxic conditions at night. PMID- 16348936 TI - Development of a rapid assimilable organic carbon method for water. AB - A rapid method for measurement of assimilable organic carbon (AOC) is proposed. The time needed to perform the assay is reduced by increasing the incubation temperature and increasing the inoculum density. The ATP luciferin-luciferase method quickly enumerates the test organisms without the need for plate count media or dilution bottles. There was no significant difference between AOC values determined with strain P17 for the ATP and plate count procedures. For strain NOX, the plate count procedure underestimated bacterial levels in some samples. Comparison of AOC values obtained by the Belleville laboratory (by the ATP technique) and the Stroud Water Research Center (by plate counts) showed that values were significantly correlated and not significantly different. The study concludes that the rapid AOC method can quickly determine the bacterial growth potential of water within 2 to 4 days. PMID- 16348937 TI - Biochemical Characterization of a Protease Involved in the Processing of a Streptomyces reticuli Cellulase (Avicelase). AB - A 36-kDa protease from Streptomyces reticuli had recently been shown to be responsible for the in vivo and in vitro processing of the 82-kDa cellulase (Avicelase) Cel-1 from S. reticuli to a 42-kDa truncated enzyme. It was induced only in the presence of Avicel, hydroxyethylcellulose, and xylan. The addition of the nonionic detergent Tween 80 to the culture medium containing Avicel as the carbon source led to a 10-fold increase in extracellular proteolytic activity. The protease, which has an isoelectric point of 3.9, was purified to homogeneity from the culture filtrate by a combination of anion-exchange and hydrophobic interaction chromatographies and was characterized biochemically. The enzyme hydrolyzed gelatin and the chromogenic substrates Azocoll, Azocasein, and Azoalbumin. Its highest activity was determined between pH 7.0 and 7.7 and at 55 degrees C. The proteolytic activity was inhibited by 1,10-phenanthroline and EDTA; however, no metal ions were detected to be associated with the protein. The protease was stable in the presence of 1 M urea and 0.01 M sodium dodecyl sulfate. The inhibitory effect of alpha-2-macroglobulin indicated an endo-mode of proteolytic cleavage. Studies with lectins and sugar analysis by mass spectroscopy indicated that the cellulase (Avicelase) Cel-1 was neither N nor O glycosylated. Its processing by the protease occurred at temperatures ranging from 30 to 55 degrees C, pH 7.5, in the presence of 2 mM dithiothreitol. PMID- 16348938 TI - Novel method for identifying bacterial mutants with reduced epiphytic fitness. AB - In order to identify novel traits involved in epiphytic colonization, a technique for the rapid identification of bacterial mutants with quantitatively different population sizes in a natural habitat based on measurements of ice nucleation activity was developed. The threshold freezing temperatures of leaves harboring different numbers of cells of ice nucleation-active Pseudomonas syringae B728a differed substantially. While few leaves containing less than about 10 cells per g (fresh weight) froze at assay temperatures of -2.75 degrees C or higher, nearly all leaves froze at these temperatures when population sizes of this strain increased to about 10 cells per g (fresh weight). Presumptive epiphytic fitness mutants could readily be identified as strains which initiated freezing in fewer leaves than did other strains within a given experiment. Most Tn5-induced mutants of strain B728a which conferred a low frequency of ice nucleation on inoculated bean leaves generally had a smaller population size than the parental strain at the time of the leaf freezing assay. The leaf freezing assay was capable of differentiating samples which varied by approximately three- to fivefold in mean bacterial population size. PMID- 16348939 TI - Characteristics of Insertional Mutants of Pseudomonas syringae with Reduced Epiphytic Fitness. AB - Random Tn5 mutagenesis was used to identify genes ir. Pseudomonas syringae which contribute to epiphytic fitness. Mutants were selected on the basis of deficiencies in epiphytic growth or survival on plants rather than deficiencies in predetermined phenotypes exhibited in culture. A sample freezing procedure was used to measure the population sizes of 5,300 mutants of P. syringae exposed to alternating wet and dry conditions on bean leaves in growth chambers. Eighty-two mutants exhibited reduced population sizes. Of these mutants, over half exhibited a reduced ability to survive the stresses associated with dry leaves, while others grew more slowly or attained reduced stationary-phase population sizes on leaves. While some epiphytic fitness mutants were altered in phenotypes that could be measured in culture, many mutants were not altered in any in vitro phenotype examined. Only three of the epiphytic fitness mutants were auxotrophs, and none had catabolic deficiencies for any of 31 organic compounds tested. Other mutants that exhibited reductions in one or more of the following were identified: motility, osmotolerance, desiccation tolerance, growth rate in batch culture, and extracellular polysaccharide production. All of the mutants retained the abilities to produce disease symptoms on the compatible host plant, bean, to incite a hypersensitive response on the non-host plant, tobacco, and to produce a fluorescent pyoverdine siderophore. PMID- 16348940 TI - Influence of the Endogenous Storage Lipid Poly-beta-Hydroxybutyrate on the Reducing Power Availability during Cometabolism of Trichloroethylene and Naphthalene by Resting Methanotrophic Mixed Cultures. AB - The role of the storage lipid poly-beta-hydroxybutyrate (PHB) in trichloroethylene transformation by methanotrophic mixed cultures was investigated. Naphthalene oxidation rates were used to assay for soluble methane monooxygenase activity. The PHB content of methanotrophic cells grown in reactors varied diurnally as well as from day to day. A positive correlation between the amount of PHB in the cells and the naphthalene oxidation rate as well as between PHB and the trichloroethylene transformation rate and capacity was found. Addition of beta-hydroxybutyrate increased the naphthalene oxidation rates significantly. PHB content in cells could be manipulated by incubation at different methane-to-nitrogen ratios. A positive correlation between the naphthalene oxidation rate and the PHB content after these incubations could be seen. Both the PHB content and the naphthalene oxidation rates decreased with time in resting methanotrophic cells exposed to oxygen. However, this decrease in the naphthalene oxidation rate cannot be explained by the decrease in the PHB content alone. Probably a deactivation of the methane monooxygenase itself is also involved. PMID- 16348941 TI - Effects of Environmental and Nutritional Factors on Production of the Polyketide Phytotoxin Coronatine by Pseudomonas syringae pv. Glycinea. AB - Pseudomonas syringae pv. glycinea PG4180 produces the polyketide phytotoxin coronatine. The effects of environmental, nutritional, and host factors on growth and coronatine production by PG4180 were examined by varying the components of a defined basal medium which contained the following nutrients per liter: glucose (10 g), NH(4)Cl (1 g), MgSO(4) . 7H(2)O (0.2 g), KH(2)PO(4) (4.1 g), K(2)HPO(4) . 3H(2)O (3.6 g), and FeCl(3) (2 muM). Bacterial growth was recorded as dry weight, and coronatine production was measured by high-performance liquid chromatography. Both growth and the quantity of coronatine synthesized were significantly affected by carbon source, nutrient levels (glucose, NH(4)Cl, phosphate, Mg, and SO(4)), amino acid supplements, and the presence of complex carbon and nitrogen sources. The yield of coronatine generally declined when conditions were varied from those in the basal medium. Coronatine production and growth were not affected when the pH was adjusted from 6.5 to 7.8. Increases in the osmolarity of the basal medium significantly decreased coronatine production without affecting growth. The addition of plant extracts, plant-derived secondary metabolites, or zinc did not affect growth or coronatine production, while the addition of millimolar levels of KNO(3) or micromolar levels of FeCl(3) significantly enhanced coronatine production. The yield of coronatine was maximized after a 7 day incubation at 18 degrees C and 280 rpm. The results of the present study were used to formulate a medium which allowed for enhanced coronatine production in nearly all strains of P. syringae tested. A rapid method for extracting coronatine from small volumes of culture supernatant was also developed. PMID- 16348942 TI - Similarity between Copper Resistance Genes from Xanthomonas campestris and Pseudomonas syringae. AB - Plasmid-borne copper resistance genes from copper-resistant strains of Xanthomonas campestris pv. vesicatoria from California, Florida, and Oklahoma shared structural similarities. A strain of X. campestris pv. campestris also contained plasmid-borne copper resistance genes similar to the resistance genes from X. campestris pv. vesicatoria. Furthermore, a region of the copper resistance genes from X. campestris pv. vesicatoria 07882 hybridized with copA, the first gene of the copper resistance operon (cop) of Pseudomonas syringae pv. tomato. A copper-inducible protein of similar size to CopA was detected by Western blot (immunoblot) analysis from the wild-type strain 07882 and from the cloned copper resistance genes of 07882 introduced into a copper-sensitive strain of X. campestris pv. vesicatoria. A low level of hybridization was observed with chromosomal DNA from other xanthomonads when the copper resistance genes from strain 07882 were used as probes. PMID- 16348943 TI - Natural Transformation of Acinetobacter calcoaceticus by Plasmid DNA Adsorbed on Sand and Groundwater Aquifer Material. AB - It is known that plasmid DNA and linear duplex DNA molecules adsorb to chemically purified mineral grains of sand and to particles of several clay fractions. It seemed desirable to examine whether plasmid DNA would also adsorb to nonpurified mineral materials taken from the environment and, particularly, whether adsorbed plasmid DNA would be available for natural transformation of bacteria. Therefore, microcosms consisting of chemically pure sea sand plus buffered CaCl(2) solution were compared with microcosms consisting of material sampled directly from a groundwater aquifer (GWA) plus groundwater (GW) with respect to the natural transformation of Acinetobacter calcoaceticus by mineral-associated DNA. The GWA minerals were mostly sand with inorganic precipitates and organic material plus minor quantities of silt and clay (illite and kaolinite). The amount of plasmid DNA which adsorbed to GWA (in GW) was about 80% of the amount which adsorbed to purified sand (in buffered CaCl(2) solution). Plasmid DNA adsorbed on sand transformed A. calcoaceticus significantly less efficiently than did plasmid DNA in solution. In contrast, the transformation by sand-adsorbed chromosomal DNA was as high as that by DNA in solution. In GWA/GW microcosms, the efficiency of transformation by chromosomal DNA was similar to that in sand microcosms, whereas plasmid transformation was not detectable. However, plasmid transformants were found at a low frequency when GWA was loaded with both chromosomal and plasmid DNA. Reasons for the low transformation efficiency of plasmid DNA adsorbed to mineral surfaces are discussed. Control experiments showed that the amounts of plasmid and chromosomal DNA desorbing from sand during incubation with a cell free filtrate of a competent cell suspension did not greatly contribute to transformation in sand microcosms, suggesting that transformation occurred by direct uptake of DNA from the mineral surfaces. Taken together, the observations suggest that plasmid DNA and chromosomal DNA fragments which are adsorbed on mineral surfaces in a sedimentary or soil habitat may be available (although with different efficiencies for the two DNA species) for transformation of a naturally competent gram-negative soil bacterium. PMID- 16348944 TI - Copper Hypersensitivity and Uptake in Pseudomonas syringae Containing Cloned Components of the Copper Resistance Operon. AB - Copper resistance in Pseudomonas syringae carrying the copABCD operon is associated with accumulation of copper in the periplasm and outer membrane, apparently as a function of the copper-binding activities of the copABC gene products. However, no specific function for copD has been determined. In this study, P. syringae cells containing copCD or copBCD cloned behind the lac promoter were hypersensitive to copper. An increased accumulation of copper was measured in cells containing several combinations of cop genes that included copC and copD. Our data suggest that CopC, a periplasmic copper-binding protein, and CopD, a probable inner membrane protein, may function together in copper uptake. PMID- 16348945 TI - Siderophore Utilization by Bradyrhizobium japonicum. AB - Bradyrhizobium japonicum USDA 110 and 61A152 can utilize the hydroxamate-type siderophores ferrichrome and rhodotorulate, in addition to ferric citrate, to overcome iron starvation. These strains can also utilize the pyoverdin-type siderophore pseudobactin St3. The ability to utilize another organism's siderophores may confer a selective advantage in the rhizosphere. PMID- 16348946 TI - Molecular Rearrangement of Longifolene by Arthrobacter ilicis T(2). AB - Arthrobacter ilicis T(2) brings about a unique type of cometabolic structural rearrangement of longifolene, a sesquiterpene, resulting in the formation of an acid. Infrared, nuclear magnetic resonance, mass spectrometry, and decoupling studies indicate that the acid product has a sativenelike structure, which is confirmed by conversion of the acid to its methyl ester and hydrocarbon. PMID- 16348947 TI - Monitoring airborne fungal spores in an experimental indoor environment to evaluate sampling methods and the effects of human activity on air sampling. AB - [This corrects the article on p. 226 in vol. 59.]. PMID- 16348948 TI - Whole-Cell Hybridization of Frankia Strains with Fluorescence- or Digoxigenin Labeled, 16S rRNA-Targeted Oligonucleotide Probes. AB - Whole-cell hybridization with non-radioactively labeled oligonucleotide probes was used to detect and identify Frankia strains in pure cultures and in nodules. Digoxigenin-labeled probes, which were detected with antibody-alkaline phosphatase conjugates, were more suitable for in situ detection of Frankia strains than fluorescent probes since the sensitivity of the former was higher and problems arising from the autofluorescence of cells and plant material were avoided. Successful detection of Frankia strains in paraformaldehyde-fixed cell material with digoxigenin-labeled oligonucleotide probes depended on pretreatments to permeabilize the cells. Specific hybridization signals on vesicles were obtained after lysozyme pretreatment (1 mg ml for 30 min at 20 degrees C). Reliable penetration of the antibody-enzyme conjugate into hyphae required additional washing with the detergent Nonidet P-40 (0.1%) and toluene (1% in ethanol) after lysozyme treatment. Identification of Frankia vesicles in nodule homogenates was possible only after the removal of the polysaccharide capsule surrounding the vesicles. Incubation with H(2)O(2) (15% in water for 1 h at room temperature) before lysozyme and detergent treatments was found to facilitate specific hybridization. No filaments or spores could be detected in nodule homogenates. This technique should be a powerful tool in the identification of Frankia isolates, in the characterization of as-yet-uncultured nodule populations, and in the confirmation of the origin of unusual Frankia isolates. PMID- 16348949 TI - Selection of xenobiotic-degrading microorganisms in a biphasic aqueous-organic system. AB - Microbial selection on mixtures of chlorinated and nonchlorinated compounds that are poorly soluble in water and/or toxic to growing microbial cells was examined in both biphasic aqueous-organic and monophasic aqueous systems. A biphasic system in which silicone oil was used as the organic phase permitted the acceleration of acclimation, leading to rapid selection and to an increase in xenobiotic compound degradation. In contrast, acclimation, selection, and degradation were very slow in the monophasic aqueous system. The variation in microbial growth rate with the degree of dispersion (i.e., dispersion at different silicone oil concentrations and agitation rates), and cell adhesion to the silicone oil indicate that the performance of the biphasic aqueous-organic system is dependent on the interfacial area between the two phases and that microbial activity is important at this interface. Therefore, the biphasic water silicone oil system could be used for microbial selection in the presence of xenobiotic compounds that are toxic and have low water solubility. PMID- 16348950 TI - Acetate Degradation at 70 degrees C in Upflow Anaerobic Sludge Blanket Reactors and Temperature Response of Granules Grown at 70 degrees C. AB - Anaerobic acetate degradation at 70 degrees C and at 55 degrees C (as a reference) was studied by running laboratory upflow anaerobic sludge blanket (UASB) reactors inoculated with mesophilic granular sludge. In UASB reactors fed with acetate-containing media (3 g of chemical oxygen demand [COD] per liter, corresponding to 47 mM acetate) approximately 50 days was needed at 70 degrees C and less than 15 days was needed at 55 degrees C to achieve an effluent COD of 500 to 700 mg/liter. In the UASB reactors at both 70 and 55 degrees C up to 90% of the COD was removed. Batch assays showed that sludges from two 70 degrees C UASB reactors, one run at a low effluent acetate concentration and the other run at a high effluent acetate concentration, exhibited slightly different responses to temperatures in the range from 37 to 70 degrees C. Both 70 degrees C sludges, as well as the 55 degrees C sludge, produced methane at temperatures of 37 to 73 degrees C. The 55 degrees C sludge exhibited shorter lag phases than the 70 degrees C sludges and higher specific methane production rates between 37 and 65 degrees C. PMID- 16348951 TI - Identification of Gremmeniella abietina Races with Random Amplified Polymorphic DNA Markers. AB - Seven random amplified polymorphic DNA (RAPD) markers amplified from four oligonucleotides (10-mers) by the polymerase chain reaction were used to distinguish between the North American and European races of Gremmeniella abietina, the causal agent of Scleroderris canker of conifers. Forty-three isolates of the pathogen from 11 different host species originating from 11 countries, states, and provinces were tested; race designation was consistent with results from immunogenic and soluble-protein assays. By using RAPD markers, it was possible to identify G. abietina races by DNA amplifications directly from fruiting bodies, thus eliminating the need to culture the fungus, as is necessary with immunogenic and soluble-protein assays. Two isolates which had been previously classified as intermediate were clearly identified as belonging to either one of the two races by using RAPD markers. No interracial hybrids were detected in our survey. Patterns of amplification products from the European race in North America were identical to patterns of European isolates, further substantiating that this is an introduced race to the North American continent. PMID- 16348952 TI - Erwinia chrysanthemi EC16 Produces a Second Set of Plant-Inducible Pectate Lyase Isozymes. AB - The enterobacterium Erwinia chrysanthemi causes soft-rot diseases involving extensive tissue maceration in a wide variety of plants and secretes multiple pectic enzymes that degrade plant cell walls and middle lamellae. An E. chrysanthemi mutant with directed deletions or insertions in genes pehX, pelX, pelA, pelB, pelC, and pelE, which encode exo-poly-alpha-d-galacturonosidase, exopolygalacturonate lyase, and four isozymes of pectate lyase, respectively, was constructed by the marker exchange of a cloned pehX::TnphoA fragment into E. chrysanthemi CUCPB5010, a Delta(pelA pelE) Delta(pelB pelC)::28bp Delta(pelX)Delta4bp derivative of strain EC16. This mutant, E. chrysanthemi CUCPB5012, no longer caused pitting in a standard pectate semisolid agar medium used to detect pectolytic activity in bacteria. Nevertheless, the mutant still macerated leaves of chrysanthemum (Chrysanthemum morifolium), although with reduced virulence. The mutant was found to produce significant pectate lyase activity in rotting chrysanthemum tissue and in minimal media containing chrysanthemum extracts or cell walls as the sole carbon source. Activity-stained, ultra-thin-layer isoelectric focusing gels revealed the presence in these preparations of several pectate lyase isozymes with pIs ranging from highly acidic to highly alkaline. Sterile culture fluids containing these isozymes were able to macerate chrysanthemum leaf tissue. Unlike the products of the pelA, pelB, pelC, and pelE genes in E. chrysanthemi EC16, these plant-inducible pectate lyase isozymes were not produced in minimal medium containing pectate. The results suggest that E. chrysanthemi produces two sets of independently regulated pectate lyase isozymes that are capable of macerating plant tissues. PMID- 16348953 TI - Plasmids pJP4 and r68.45 Can Be Transferred between Populations of Bradyrhizobia in Nonsterile Soil. AB - IncP plasmid r68.45, which carries several antibiotic resistance genes, and IncP plasmid pJP4, which contains genes for mercury resistance and 2,4 dichlorophenoxyacetic acid degradation, were evaluated for their ability to transfer to soil populations of rhizobia. Transfer of r68.45 was detected in nonsterile soil by using Bradyrhizobium japonicum USDA 123 as the plasmid donor and several Bradyrhizobium sp. strains as recipients. Plasmid transfer frequencies ranged up to 9.1 x 10 in soil amended with 0.1% soybean meal and were highest after 7 days with strain 3G4b4-RS as the recipient. Transconjugants were detected in 7 of 500 soybean nodules tested, but the absence of both parental strains in these nodules suggests that plasmid transfer had occurred in the soil, in the rhizosphere, or on the root surface. Transfer of degradative plasmid pJP4 was also evaluated in nonsterile soil by using B. japonicum USDA 438 as the plasmid donor and several Bradyrhizobium sp. strains as recipients. Plasmid pJP4 was transferred only when strains USDA 110-ARS and 3G4b4-RS were the recipients. The plasmid transfer frequency was highest for strain 3G4b4-RS (up to 7.4 x 10). Mercury additions to soil, ranging from 10 to 50 mug/g of soil, did not affect population levels of parental strains or the plasmid transfer frequency. PMID- 16348954 TI - Adhesion of Nongerminated Botrytis cinerea Conidia to Several Substrata. AB - Conidia of the plant pathogenic fungus Botrytis cinerea adhered to tomato cuticle and to certain other substrata immediately upon hydration. This immediate adhesion occurred with both living and nonliving conidia. Adhesion was not consistently influenced by several lectins, sugars, or salts or by protease treatment, but it was strongly inhibited by ionic or nonionic detergents. With glass and oxidized polyethylene, substrata whose surface hydrophobicities could be conveniently varied, there was a direct relationship between water contact angle and percent adhesion. Immediate adhesion did not involve specific conidial attachment structures, although the surfaces of attached conidia were altered by contact with a substratum. Freshly harvested conidia were very hydrophobic, with more than 97% partitioning into the organic layer when subjected to a phase distribution test. Percent adhesion of germinated conidia was larger than that of nongerminated conidia. Evidence suggests that immediate adhesion of conidia of B. cinerea depends, at least in part, on hydrophobic interactions between the conidia and substratum. PMID- 16348955 TI - Extracellular Enzyme Production and Synthetic Lignin Mineralization by Ceriporiopsis subvermispora. AB - The ability of the white rot fungus Ceriporiopsis subvermispora to mineralize C synthetic lignin was studied under different culture conditions, and the levels of two extracellular enzymes were monitored. The highest mineralization rates (28% after 28 days) were obtained in cultures containing a growth-limiting amount of nitrogen source (1.0 mM ammonium tartrate); under this condition, the levels of manganese peroxidase (MnP) and laccase present in the culture supernatant solutions were very low compared with cultures containing 10 mM of the nitrogen source. In contrast, cultures containing a limiting concentration of the carbon source (0.1% glucose) showed low levels of both enzymes and also very low mineralization rates compared with cultures containing 1% glucose. Cultures containing 11 ppm of Mn(II) showed a higher rate of mineralization than those containing 0.3 or 40 ppm of this cation. Levels of MnP and laccase were higher when 40 ppm of Mn(II) was used. Mineralization rates were slightly higher in cultures flushed daily with oxygen, whereas laccase levels were lower and MnP levels were approximately the same as in cultures maintained under an air atmosphere. The presence of 0.4 mM veratryl alcohol reduced both mineralization rates and MnP levels, without affecting laccase levels. Lignin peroxidase activity was not detected under any condition. Addition of purified lignin peroxidase to the cultures in the presence or absence of veratryl alcohol did not enhance mineralization rates significantly. PMID- 16348956 TI - Studies of the Physiological and Genetic Basis of Acid Tolerance in Rhizobium leguminosarum biovar trifolii. AB - Acid-tolerant Rhizobium leguminosarum biovar trifolii ANU1173 was able to grow on laboratory media at a pH as low as 4.5. Transposon Tn5 mutagenesis was used to isolate mutants of strain ANU1173, which were unable to grow on media at a pH of less than 4.8. The acid-tolerant strain ANU1173 maintained a near-neutral intracellular pH when the external pH was as low as 4.5. In contrast, the acid sensitive mutants AS25 and AS28 derived from ANU1173 had an acidic intracellular pH when the external pH was less than 5.5. The acid-sensitive R. leguminosarum biovar trifolii ANU794, which was comparatively more sensitive to low pH than mutants AS25 and AS28, showed a more acidic internal pH than the two mutants when the three strains were exposed to medium buffered at a pH of less than 5.5. The two acid-sensitive mutants had an increased membrane permeability to protons but did not change their proton extrusion activities. However, the acid-sensitive strain ANU794 exhibited both a higher membrane permeability to protons and a lower proton extrusion activity compared with the acid-tolerant strain ANU1173. DNA hybridization analysis showed that mutants AS25 and AS28 carried a single copy of Tn5 located in 13.7-kb (AS25) and 10.0-kb (AS28) EcoRI DNA fragments. The wild-type DNA sequences spanning the mutation sites of mutants AS25 and AS28 were cloned from genomic DNA of strain ANU1173. Transfer of these wild-type DNA sequences into corresponding Tn5-induced acid-sensitive mutants, respectively, restored the mutants to their acid tolerance phenotypes. Mapping studies showed that the AS25 locus was mapped to a 5.6-kb EcoRI-BamHI megaplasmid DNA fragment, whilst the AS28 locus was located in an 8.7-kb BglII chromosomal DNA fragment. PMID- 16348957 TI - Lipopolysaccharides as Determinants of Serological Variability in Pseudomonas corrugata. AB - The variation in biochemical and serological features of 128 isolates of Pseudomonas corrugata has been studied with 56 isolates from Spain and 72 isolates from other countries. Isolates were analyzed with common diagnostic tests and with the AP150CHE system. Variability among isolates for some standard tests usually listed as positive or negative for this species, such as arginine dihydrolase and gelatin hydrolysis, lipase and lecithinase activities, pigment production, and wrinkled colony morphology, was observed. Three antisera were raised against the type strain and two Spanish isolates from tomato and pepper plants. Serological reactions were studied by indirect immunofluorescence and indirect enzyme-linked immunosorbent assay. Eighty-three isolates reacted with a single antiserum, 6 reacted with two antisera, and none reacted with three antisera. Thirty-nine isolates did not react with any of the three antisera. These results suggest that serology will not be a useful method for routine diagnosis of P. corrugata unless common antigens can be identified. Electrophoresis and immunoelectrotransfer were used to study the antigens involved. Each antiserum reacted with whole-cell lysates, giving two common bands for P. corrugata isolates and other Pseudomonas species and a ladder-like pattern characteristic of lipopolysaccharides (LPS). Common bands were not observed after proteinase K treatment. More than 10 LPS patterns were distinguished in 98 isolates after silver staining of polyacrylamide gels. There was no correlation between the geographical origin or host of the isolates and the LPS patterns. A correlation between LPS groups and serological reaction was observed. PMID- 16348958 TI - Impact of bacterial biomass on contaminant sorption and transport in a subsurface soil. AB - The objective of this study was to investigate the impact of bacterial biomass on the sorption and transport of three solutes (quinoline, naphthalene, and Ca) in a subsurface soil. Miscible displacement techniques were employed to measure sorption and transport of the above compounds during steady, saturated water flow in sterile and/or bacterium-inoculated soil columns. The soil was inoculated with either a quinoline-degrading bacterium, Pseudomonas sp. 3N3A isolate, or its mutant isolate, B53, which does not degrade quinoline. In soil columns inoculated with the B53 and 3N3A isolates, quinoline sorption was reduced by about 60 and 20%, respectively. In contrast, Ca sorption was minimally reduced, which indicated that biomass did not significantly alter the cation-exchange capacity of the soil. Biomass impacts on sorption were solute specific, even when the sorption mechanism for both quinoline and Ca was similar. Thus, the differential response is attributed to biomass-induced changes in quinoline speciation; an increase in pH at the sorbent-water interface would result in a larger proportion of the neutral species and a decrease in sorption. Sorption of naphthalene was reduced by about 30%, which was attributed to accessibility of hydrophobic regions. Minimal biosorption of all solutes indicated negligible biofacilitated transport. Alteration of the soil surfaces upon addition of bacterial biomass reduced sorption of quinoline and naphthalene, thereby enhancing transport. PMID- 16348959 TI - Biotinylation of Bacillus thuringiensis Insecticidal Crystal Proteins. AB - Biotinylation of Bacillus thuringiensis insecticidal crystal proteins (ICPs) was evaluated for its potential use in an alternative ICP screening method and in the characterization of ICP receptors. In vivo biological activity of CryIA(b), as inferred from bioassays with Manduca sexta and Ostrinia nubilalis and from histopathological effects on O. nubilalis midgut cells induced by force feeding, was not affected by biotinylation at moderate biotinylation ratios. A competitive radioreceptor assay showed that there was only a minor reduction in binding affinity of biotin-labeled CryIA(b) for M. sexta brush border membrane vesicles. On midgut tissue sections, the binding pattern along the midgut epithelium and the staining intensity of biotinylated ICPs detected with streptavidin-enzyme conjugate were virtually identical to the binding pattern and staining intensity of native CryIA(b) detected with antibodies. The specificity of biotinylated ICP binding to larval midgut tissue was demonstrated by performing homologous competition experiments. The relationship between different ICP receptor types in Plutella xylostella, as inferred from radioligand binding studies, was confirmed by the results of heterologous competition experiments performed with biotinylated and native ICPs. PMID- 16348960 TI - Two Different Bacillus thuringiensis Delta-Endotoxin Receptors in the Midgut Brush Border Membrane of the European Corn Borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Pyralidae). AB - Binding of three Bacillus thuringiensis insecticidal crystal proteins (ICPs) to the midgut epithelium of Ostrinia nubilalis larvae was characterized by performing binding experiments with both isolated brush border membrane vesicles and gut tissue sections. Our results demonstrate that two independent ICP receptors are present in the brush border of O. nubilalis gut epithelium. From competition binding experiments performed with I-labeled and native ICPs it was concluded that CryIA(b) and CryIA(c) are recognized by the same receptor. An 11 fold-higher binding affinity of CryIA(b) for this receptor correlated with a 10 fold-higher toxicity of this ICP compared with CryIA(c). The CryIB toxin did not compete for the binding site of CryIA(b) and CryIA(c). Immunological detection of ingested B. thuringiensis ICPs on gut sections of O. nubilalis larvae revealed binding only along the epithelial brush border membrane. CryID and CryIE, two ICPs that are not toxic to O. nubilalis, were not bound to the apical microvilli of gut epithelial cells. In vitro binding experiments performed with native and biotinylated ICPs on tissue sections confirmed the correlation between ICP binding and toxicity. Moreover, by performing heterologous competition experiments with biotinylated and native ICPs, it was confirmed that the CryIB receptor is different from the receptor for CryIA(b) and CryIA(c). Retention of activated crystal proteins by the peritrophic membrane was not correlated with toxicity. Furthermore, it was demonstrated that CryIA(b), CryIA(c), and CryIB toxins interact in vitro with the epithelial microvilli of Malpighian tubules. In addition, CryIA(c) toxin also adheres to the basement membrane of the midgut epithelium. PMID- 16348961 TI - Biometric Study of Acetoin Production in Hanseniaspora guilliermondii and Kloeckera apiculata. AB - Gas chromatographic analysis by direct injection of samples yielded quantitative data on acetoin content. Ninety-six strains of Hanseniaspora guilliermondii and Kloeckera apiculata were investigated for the ability to produce acetoin in synthetic medium and in must. High-level production of acetoin was found to be a characteristic of both species. In synthetic medium, the two species were not significantly different with respect to sugar utilization and ethanol or acetoin production. In grape must, the two species were significantly different (P = 0.001) in acetoin production and K. apiculata exhibited a significantly negative correlation between acetoin production and either sugar consumption or ethanol production. Use of selected apiculate yeasts in mixed cultures with Saccharomyces cerevisiae seems promising for optimization of wine bouquet. PMID- 16348962 TI - Indigenous bacteria in hemolymph and tissues of marine bivalves at low temperatures. AB - Hemolymph and soft tissues of Pacific oysters (Crassostrea gigas) kept in sand filtered seawater at temperatures between 1 and 8 degrees C were normally found to contain bacteria, with viable counts (CFU) in hemolymph in the range 1.4 x 10 to 5.6 x 10 bacteria per ml. Pseudomonas, Alteromonas, Vibrio, and Aeromonas organisms dominated, with a smaller variety of morphologically different unidentified strains. Hemolymph and soft tissues of horse mussels (Modiolus modiolus), locally collected from a 6- to 10-m depth in the sea at temperatures between 4 and 6 degrees C, also contained bacteria. The CFU in horse mussel hemolymph was of the same magnitude as that in oysters (mean, 2.6 x 10), and the bacterial flora was dominated by Pseudomonas (61.3%), Vibrio (27.0%), and Aeromonas (11.7%) organisms. In soft tissues of horse mussels, a mean CFU of 2.9 x 10 bacteria per g was found, with Vibrio (38.5%), Pseudomonas (33.0%), and Aeromonas (28.5%) constituting the major genera. After the challenge of oysters in seawater at 4 degrees C to the psychrotrophic fish pathogen Vibrio salmonicida (strains NCIMB 2245 from Scotland and TEO 84001 from Norway) and a commensal Aeromonas sp. isolated from oysters, the viable count in hemolymph increased 1,000-fold to about 10 bacteria per ml. In soft tissues, about a 1,000-fold increase in CFU to 6 x 10 was observed. V. salmonicida NCIMB 2245 invaded hemolymph and soft tissues after 14 days and dominated these compartments after 41 days, whereas strain TEO 84001 did not invade soft tissues to the same extent. Challenge with V. salmonicida NCIMB 2245 resulted in 100% mortality, whereas about 50% of the oysters survived challenge with the Norwegian strain, TEO 84001. The commensal Aeromonas sp. invaded hemolymph and soft tissues and caused 100% mortality. Oyster hemolymph contained agglutinins for Vibrio anguillarum but not for V. salmonicida, whereas we did not find agglutinins for either of these bacteria in horse mussels. Agglutinins for horse and human erythrocytes were found in hemolymph from both animals. We found no differences in agglutinin titers in oysters from different Norwegian locations, and long-term challenge with bacteria in seawater did not result in changes of agglutinin activity. These studies demonstrate that bacteria exist in hemolymph and soft tissues of marine bivalves at temperatures below 8 degrees C. Increased bacterial numbers in seawater at 4 degrees C result in augmented invasion of bacteria in hemolymph and soft tissues. V. salmonicida, a bacterium pathogenic for fish at low temperatures, invades bivalve hemolymph and soft tissues, and thus bivalves may serve as a reservoir for pathogens of fish at low seawater temperatures. PMID- 16348963 TI - Effects of Kraft Pulp and Lignin on Trametes versicolor Carbon Metabolism. AB - The white rot basidiomycete Trametes (Coriolus) versicolor can substantially increase the brightness and decrease the lignin content of washed, unbleached hardwood kraft pulp (HWKP). Monokaryotic strain 52J was used to study how HWKP and the lignin in HWKP affect the carbon metabolism and secretions of T. versicolor. Earlier work indicated that a biobleaching culture supernatant contained all components necessary for HWKP biobleaching and delignification, but the supernatant needed frequent contact with the fungus to maintain these activities. Thus, labile small fungal metabolites may be the vital biobleaching system components renewed or replaced by the fungus. Nearly all of the CO(2) evolved by HWKP-containing cultures came from the added glucose, indicating that HWKP is not an important source of carbon or energy during biobleaching. Carbon dioxide appeared somewhat earlier in the absence of HWKP, but the culture partial O(2) pressure was little affected by the presence of pulp. The presence of HWKP in a culture markedly increased the culture's production of a number of acidic metabolites, including 2-phenyllactate, oxalate, adipate, glyoxylate, fumarate, mandelate, and glycolate. Although the total concentration of these pulp-induced metabolites was only 4.3 mM, these compounds functioned as effective manganese complexing agents for the manganese peroxidase-mediated oxidation of phenol red, propelling the reaction at 2.4 times the rate of 50 mM sodium malonate, the standard chelator-buffer. The presence of HWKP in a culture also markedly stimulated fungal secretion of the enzymes manganese peroxidase, cellulase, and cellobiose-quinone oxidoreductase, but not laccase (phenol oxidase) or lignin peroxidase. PMID- 16348964 TI - Physical characterization and quantification of bacteria by sedimentation field flow fractionation. AB - Studies in microbial ecology require accurate measures of cell number and biomass. Although epifluorescence microscopy is an accepted and dependable method for determining cell numbers, current methods of converting biovolume to biomass are error prone, tedious, and labor-intensive. This paper describes a technique with sedimentation field-flow fractionation to enumerate bacteria and determine their density, size, and mass. Using cultured cells of different shapes and sizes, we determined optimum values for separation run parameters and sample handling procedures. The technique described can separate and detect 4', 6 diamidino-2-phenylindole-stained cells and generate a fractogram from which cell numbers and their size or mass distribution can be calculated. A direct method for estimating bacterial biomass (dry organic matter content) which offers distinct advantages over present methods for calculating biomass has been developed. PMID- 16348965 TI - Presence of a Vanadium Nitrogenase in Azotobacter paspali. AB - There have been no previous studies on the genetics of Azotobacter paspali, an aerobic bacterium which forms a highly specific diazotrophic association with Bahia grass (Paspalum notatum). We constructed A. paspali strains defective in the molybdenum nitrogenase so that alternative N(2)ases could be studied. The cosmid vector pTBE and genomic DNA fragments ( approximately 50 kb) of A. paspali ATCC 23367 were used to construct a gene library in Escherichia coli. Recombinant cosmids containing sequences homologous to molybdenum nitrogenase nifDK structural genes were identified by hybridization. A 2.9-kb fragment bearing the putative nifDK genes of A. paspali was subcloned and mutagenized in vitro by the insertion of a kanamycin resistance gene cassette. The mutation was recombined into the chromosome of A. paspali with the suicide vector pCU101. One resultant mutant strain, AP2, was incapable of diazotrophic growth in a molybdenum containing medium (Nif) without vanadium but grew well in a molybdenum-deficient medium with vanadium. The nitrogenase system in AP2 reduced acetylene to ethylene and produced ethane as 2.4% of the total products. Molybdenum levels as low as 10 nM prevented the diazotrophic growth of AP2, even in the presence of vanadium at levels up to 10 muM. These results are consistent with the existence of a vanadium nitrogenase system in A. paspali. PMID- 16348966 TI - Effect of Initial Oxygen Concentration on Diacetyl and Acetoin Production by Lactococcus lactis subsp. lactis biovar diacetylactis. AB - The production of aroma compounds (acetoin and diacetyl) in fresh unripened cheese by Lactococcus lactis subsp. lactis biovar diacetylactis CNRZ 483 was studied at 30 degrees C at different initial oxygen concentrations (0, 21, 50, and 100% of the medium saturation by oxygen). Regardless of the initial O(2) concentration, maximal production of these compounds was reached only after all the citrate was consumed. Diacetyl and acetoin production was 0.01 and 2.4 mM, respectively, at 0% oxygen. Maximum acetoin concentration reached 5.4 mM at 100% oxygen. Diacetyl production was increased by factors of 2, 6, and 18 at initial oxygen concentrations of 21, 50, and 100%, respectively. The diacetyl/acetoin concentration ratio increased linearly with initial oxygen concentration: it was eight times higher at 100% (3.3%) than at 0% oxygen (0.4%). The effect of oxygen on diacetyl and acetoin production was also shown with other lactococci. At 0% oxygen, specific activity of alpha-acetolactate synthetase (0.15 U/mg) and NADH oxidase (0.04 U/mg) was 3.6 and 5.4 times lower, respectively, than at 100% oxygen. The increasing alpha-acetolactate synthetase activity in the presence of oxygen would explain the higher production of diacetyl and acetoin. The NADH oxidase activity would replace the role of the lactate dehydrogenase, diacetyl reductase, and acetoin reductase in the reoxidation of NADH, allowing accumulation of these two aroma compounds. PMID- 16348967 TI - 5-hydroxyquinoline-2-carboxylic Acid, a dead-end metabolite from the bacterial oxidation of 5-aminonaphthalene-2-sulfonic Acid. AB - 5-Aminonaphthalene-2-sulfonate (5A2NS) is converted by strain BN6 into 5 hydroxyquinoline-2-carboxylate (5H2QC). The authenticity of this new compound is confirmed by nuclear magnetic resonance and mass spectrometry. Its formation is explained by a spontaneous cyclization of the hypothetical metabolite 6'-amino-2' hydroxybenzalpyruvate. The formation of 5H2QC as a dead-end product of 5A2NS prevents NADH regeneration so that 5A2NS oxidation is limited by the internal NADH pool. PMID- 16348968 TI - Presence of Culturable Bacteria in Cocoons of the Earthworm Eisenia fetida. AB - Viable bacteria were found to coexist with developing embryos in egg capsules (cocoons) of the earthworm Eisenia fetida. Earthworms were reared under standardized conditions, and bacterial densities were measured in distinct batches of cocoons collected weekly for 10 weeks. Cocoons weighing 12 mg contained a mean viable bacterial population of approximately 10 CFU/g of cocoons. No difference was found in viable counts obtained from cocoons incubated at 15 degrees C and cocoons incubated at 24 degrees C. Viable bacterial numbers increased with cocoon age, while acridine orange direct counts of microbial cells were stable at approximately 10 cells per g of cocoons. Bacteria isolated from cocoons were used to develop antisera in rabbits for the production of strain specific fluorescent antibodies. Fluorescent antibody and selective plating techniques were used to monitor populations of these bacteria in earthworm bedding and to determine whether cocoons acquire bacteria from the environment in which they are formed. Cocoon isolates were readily recovered from cocoons formed in inoculated bedding at densities of 10 CFU/g of cocoons. Bradyrhizobium japonicum USDA 110 and UMR 161 added to bedding were also recovered from cocoons, but at lower densities than cocoon isolates. Escherichia coli K-12(pJP4) inoculum was recovered from bedding but not from cocoons. The bacterial complement of Eisenia fetida cocoons is affected by inoculation of selected bacterial isolates in the worm growth environment. PMID- 16348969 TI - Ethylene Production by Root Nodules and Effect of Ethylene on Nodulation in Glycine max. AB - Nodulated soybean roots produced more ethylene and contained more 1 aminocyclopropane-1-carboxylic acid than uninoculated roots. Nodules produced more ethylene and contained more 1-aminocyclopropane-1-carboxylic acid per gram of material than roots. Almost all of the ethylene produced by the nodules was produced by the plant fractions of the nodules. Ethylene, at physiological concentrations, did not inhibit nodulation in soybeans. PMID- 16348970 TI - Quantitative Determination of the Spatial Distribution of Nitrosomonas europaea and Nitrobacter agilis Cells Immobilized in kappa-Carrageenan Gel Beads by a Specific Fluorescent-Antibody Labelling Technique. AB - A novel technique, combining labelling and stereological methods, for the determination of spatial distribution of two microorganisms in a biofilm is presented. Cells of Nitrosomonas europaea (ATCC 19718) and Nitrobacter agilis (ATCC 14123) were homogeneously distributed in a kappa-carrageenan gel during immobilization and allowed to grow out to colonies. The gel beads were sliced in thin cross sections after fixation and embedding. A two-step labelling method resulted in green fluorescent colonies of either N. europaea or N. agilis in the respective cross sections. The positions and surface areas of the colonies of each species were determined, and from that a biomass volume distribution for N. europaea and N. agilis in kappa-carrageenan gel beads was estimated. This technique will be useful for the validation of biofilm models, which predict such biomass distributions. PMID- 16348971 TI - Occurrence of Vibrio alginolyticus in Ligurian Coast Rock Pools (Tyrrhenian Sea, Italy) and Its Association with the Copepod Tigriopus fulvus (Fisher 1860). AB - A study of heterotrophic bacteria and vibrios adhering to the copepod Tigriopus fulvus, which lives in Ligurian coast rock pools (Tyrrhenian Sea), was carried out from November 1990 to October 1991. Heterotrophic bacteria, which were always found both free in the water and bound to the T. fulvus organisms, showed a correlation with water temperature and salinity. Vibrio alginolyticus was found free in the water and bound to T. fulvus surfaces during the warmest months. Temperature is the main factor influencing the presence of V. alginolyticus in the rock pool. Attachment of this microorganism to the copepod provides a mechanism for its extended geographic distribution. PMID- 16348972 TI - Influence of pH and Temperature on Enumeration of Cellulose- and Hemicellulose Degrading Thermophilic Anaerobes in Neutral and Alkaline Icelandic Hot Springs. AB - Cellulose- and hemicellulose-degrading thermophilic anaerobes were enumerated in biomat samples of various temperatures from two different hot springs in the Hverageroi area of Iceland: one spring had a pH near 7, the second had a pH near 9. The most-probable-number technique was used for enumeration of bacteria in the samples, with media at many different temperatures (37 to 90 degrees C) and two pH values (7 and 9). There were generally more xylan-degrading then cellulose utilizing organisms in both environments. There was no growth at 80 degrees C in the neutral spring or at 37 degrees C in the alkaline spring. However, there were large numbers of both types of organisms in the alkaline spring at 80 degrees C and in the neutral spring at 37 degrees C. No cultures grew from the most probable-number tubes inoculated with the Hverageroi samples and incubated at 90 degrees C or with media at pH 9. However, xylan-degrading cultures at 70 degrees C were enriched at pH 9 with samples from some other Icelandic hot springs. PMID- 16348973 TI - Transduction of a Plasmid Carrying the Cohesive End Region from Lactococcus lactis Bacteriophage PhiLC3. AB - Plasmids carrying the cohesive end region from temperate lactococcal bacteriophage PhiLC3 could be packaged in vivo by PhiLC3 and transduced into its host strain, Lactococcus lactis subsp. cremoris NCDO 1201. The transduction frequencies were between 10 and 10 transducing particles per PFU, depending on the size of the phage DNA insert. This transduction system is limited to only certain lactococcal strains. The PhiLC3 cohesive site region (cos) appears to play an important role in plasmid transduction. PMID- 16348974 TI - Polyvinylpyrrolidone-agarose gel electrophoresis purification of polymerase chain reaction-amplifiable DNA from soils. AB - This communication describes a modification of agarose gel electrophoresis to provide a rapid and simple method for the purification of polymerase chain reaction-amplifiable DNA from soil. This modification is to add polyvinylpyrrolidone to the agarose gel. The polyvinylpyrrolidone addition retards the electrophoretic mobility of denaturing phenolic compounds so that they do not comigrate with nucleic acids. PMID- 16348975 TI - Effect of temperature on Fatty Acid composition of a white thermus strain. AB - A white Thermus sp. strain, NCIMB 11245, showed high levels of anteiso C(17:0), anteiso C(17:1), normal C(16:1), and iso C(16:0) with low levels of iso C(15:0) + iso C(17:0) in comparison to yellow-pigmented strains. The fatty acid composition may be associated with precursor metabolism or the absence of carotene pigmentation. PMID- 16348976 TI - Neutral red assay for measurement of quantitative vero cell cytotoxicity. AB - A neutral red assay involving Vero cells was used to quantitate the cytotoxic activity of verotoxins (VT) produced by Escherichia coli and to investigate changes in titer caused by altering the composition of the cell culture medium. Three variations on medium 199 were investigated: one involved supplementing the medium with 5% fetal bovine serum (FBS), a second was the use of serum-free (SF) medium that contained 5% bovine serum albumin and 5 mug of fibronectin per ml, and the third involved the use of 4% Ultroser G, a commercial serum replacement. The level of cytotoxicity varied markedly with the type of VT and with the medium that was used. For VT1, there was no difference in cytotoxicity between medium with 5% FBS and SF medium, but cytotoxicity was reduced more than 100-fold in medium containing Ultroser G compared with cytotoxicity in the other media. For VT2, VT2v, and VTe, there was a slight reduction in cytotoxicity in medium containing 4% Ultroser G and a more marked reduction in SF medium compared with cytotoxicity in medium containing 5% FBS. PMID- 16348977 TI - Mineral Products of Pyrrhotite Oxidation by Thiobacillus ferrooxidans. AB - The biological leaching of pyrrhotite (Fe(1-x)S) by Thiobacillus ferrooxidans was studied to characterize the oxidation process and to identify the mineral weathering products. The process was biphasic in that an initial phase of acid consumption and decrease in redox potential was followed by an acid-producing phase and an increase in redox potential. Elemental S was one of the first products of pyrrhotite degradation detected by X-ray diffraction. Pyrrhotite oxidation also yielded K-jarosite [KFe(3)(SO(4))(2)(OH)(6)], goethite (alpha FeOOH), and schwertmannite [Fe(8)O(8)(OH)(6)SO(4)] as solid-phase products. Pyrrhotite was mostly depleted after 14 days, whereas impurities in the form of pyrite (cubic FeS(2)) and marcasite (orthorhombic FeS(2)) accumulated in the leach residue. PMID- 16348978 TI - Method for Investigation of Competition between Bacteria as a Function of Three Environmental Factors Varied Simultaneously. AB - Competition between microorganisms as affected by temperature, pH, and the sodium chloride (NaCl) concentration was investigated by selective replication from gradient plates. Salmonella typhimurium was inhibited by Pseudomonas putida at 20 and 23 degrees C but not 30 and 35 degrees C. P. putida no longer grew at the extremes of pH and NaCl concentration, particularly at 30 and 35 degrees C. PMID- 16348979 TI - Long-Term Production of Ergot Peptides by Immobilized Claviceps purpurea in Semicontinuous and Continuous Culture. AB - The semicontinuous and continuous production of pharmaceutically useful ergot peptides with immobilized Claviceps purpurea could be demonstrated. A key aspect was the presence of high concentrations of CaCl(2) (96.9 mM) to give marked prolongation of the productive phase, and cultivation in a bubble column reactor became possible. Restriction of the phosphate supply avoided an otherwise problematic massive increase of outgrowing hyphae. PMID- 16348980 TI - Production of Monoclonal Antibodies against the Major Capsid Protein of the Lactococcus Bacteriophage ul36 and Development of an Enzyme-Linked Immunosorbent Assay for Direct Phage Detection in Whey and Milk. AB - The only major structural protein (35 kDa) of the lactococcal small isometric headed bacteriophage ul36, a member of the P335 species, was isolated from a preparative sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Monoclonal antibodies (MAbs) were raised against the denatured 35-kDa protein. Six MAbs were selected and characterized. Western blots (immunoblots) showed that all MAbs recognized the 35 kDa but also a 45 kDa that is in lower concentration in the phage structure. Binding inhibition assays identified five families of MAbs that recognized nonoverlapping epitopes of the 35- and 45-kDa proteins. Immunoelectron microscopy showed that these two proteins are localized within the phage head, therefore indicating that the 35 kDa is a major capsid protein of ul36 and that the 45 kDa is a minor capsid protein. With two MAbs, a sandwich enzyme-linked immunosorbent assay (ELISA) was developed for direct detection of lactococcal phages in whey and milk samples. Whey and milk components, however, interfered with the conduct of the assay. Partial denaturation of milk samples by heat treatment in the presence of SDS and beta-mercaptoethanol removed the masking effect and increased the sensitivity of the assay by 100-fold. With the method used here, 10 PFU/ml were detected by the ELISA within 2 h without any steps to enrich or isolate bacteriophages. PMID- 16348981 TI - Kinetics of bacterial growth on chlorinated aliphatic compounds. AB - With the pure bacterial cultures Ancylobacter aquaticus AD20 and AD25, Xanthobacter autotrophicus GJ10, and Pseudomonas sp. strain AD1, Monod kinetics was observed during growth in chemostat cultures on 1,2-dichloroethane (AD20, AD25, and GJ10), 2-chloroethanol (AD20 and GJ10), and 1,3-dichloro-2-propanol (AD1). Both the Michaelis-Menten constants (K(m)) of the first catabolic (dehalogenating) enzyme and the Monod half-saturation constants (K(s)) followed the order 2-chloroethanol, 1,3-dichloro-2-propanol, epichlorohydrin, and 1,2 dichloroethane. The K(s) values of strains GJ10, AD20, and AD25 for 1,2 dichloroethane were 260, 222, and 24 muM, respectively. The low K(s) value of strain AD25 was correlated with a higher haloalkane dehalogenase content of this bacterium. The growth rates of strains AD20 and GJ10 in continuous cultures on 1,2-dichloroethane were higher than the rates predicted from the kinetics of the haloalkane dehalogenase and the concentration of the enzyme in the cells. The results indicate that the efficiency of chlorinated compound removal is indeed influenced by the kinetic properties and cellular content of the first catabolic enzyme. The cell envelope did not seem to act as a barrier for permeation of 1,2 dichloroethane. PMID- 16348982 TI - Effect of X-Prolyl Dipeptidyl Aminopeptidase Deficiency on Lactococcus lactis. AB - The genetic determinant (pepXP) of an X-prolyl dipeptidyl aminopeptidase (PepXP) has recently been cloned and sequenced from both Lactococcus lactis subsp. cremoris (B. Mayo, J. Kok, K. Venema, W. Bockelmann, M. Teuber, H. Reinke, and G. Venema, Appl. Environ. Microbiol. 57:38-44, 1991) and L. lactis subsp. lactis (M. Nardi, M.-C. Chopin, A. Chopin, M.-M. Cals, and J.-C. Gripon, Appl. Environ. Microbiol. 57:45-50, 1991). To examine the possible role of the enzyme in the breakdown of caseins required for lactococci to grow in milk, integration vectors have been constructed and used to specifically inactivate the pepXP gene. After inactivation of the gene in L. lactis subsp. lactis MG1363, which is Lac and Prt, the Lac Prt determinants were transferred by conjugation by using L. lactis subsp. lactis 712 as the donor. Since growth of the transconjugants relative to the PepXP strains was not retarded in milk, it was concluded that PepXP is not essential for growth in that medium. It was also demonstrated that the open reading frame ORF1, upstream of pepXP, was not required for PepXP activity in L. lactis. A marked difference between metenkephalin degradation patterns was observed after incubation of this pentapeptide with cell extracts obtained from wild-type lactococci and pepXP mutants. Therefore, altered expression of the pepXP-encoded general dipeptidyl aminopeptidase activity may change the peptide composition of fermented milk products. PMID- 16348983 TI - Use of an Exotic Carbon Source To Selectively Increase Metabolic Activity and Growth of Pseudomonas putida in Soil. AB - Respiration and growth of Pseudomonas putida PpG7, containing catabolic plasmid NAH7, was determined in three agricultural field soils amended with the carbon source salicylate. The addition of salicylate to soil significantly increased the population of PpG7. However, there was a lack of relationship between microbial numbers and activity as determined by evolution of CO(2). In soils containing 30 to 1,500 mug of salicylate per g, metabolic activities of PpG7 peaked between 18 and 42 h and population densities increased approximately 10-to 10-fold. However, the metabolic activity of PpG7 rapidly declined after salicylate was utilized, whereas peak population densities were maintained for the duration of the experiments (5 to 7 days). Thus, elevated population densities of PpG7 were represented by inactive cells. Soil type had only minor effects on respiration rates or growth curves of PpG7 when amended with comparable concentrations of salicylate. Respiration and growth rates were optimal at concentrations between 300 and 1,000 mug of salicylate per g in the test soils. At 1,500 to 2,500 mug/g, respiration and growth of PpG7 were initially suppressed, but after a short lag time both attained levels similar to or greater than those resulting from the use of lower concentrations of salicylate. The culturing of PpG7 on a salicylate amended medium to induce salicylate-degradative enzymes did not affect the lag time before utilization of salicylate in soil. Although PpG7 competed well with fungi for the substrate, suppression of fungal populations with cycloheximide resulted in significantly increased population densities of PpG7 in two of three soils amended with salicylate. The beneficial activities of bacteria in soil are discussed in relation to population density, population metabolic activity, and selective carbon source utilization. PMID- 16348984 TI - Enhancement of Population Densities of Pseudomonas putida PpG7 in Agricultural Ecosystems by Selective Feeding with the Carbon Source Salicylate. AB - Sodium salicylate (1,000 mug/ml) was delivered through a drip irrigation system to agricultural field soils planted to tomato and infested with Pseudomonas putida PpG7, the host of the salicylate catabolic plasmid NAH7. In nonfumigated soils infested with approximately 10 CFU of PpG7 per g in the top 30 cm, population densities were increased up to 112-fold within 14 days of the initial application of salicylate compared with the densities in the respective nonamended soils. Mean season-long population densities of PpG7 in the top 30 cm of soil were significantly increased (P < 0.01) from 216 CFU/g in nonamended soils to 1,370 CFU/g in salicylate-amended soils. In the respective rhizosphere soils, mean population densities of PpG7 were significantly increased (P < 0.01) from 92 to 2,066 CFU/cm of root. Soil fumigation interacted (P < 0.01) with salicylate amendment and further increased the mean population densities of PpG7 in nonrhizosphere soil by an additional 5,689 CFU/g of soil. This fumigation effect was not detected in rhizosphere soils. The effect of salicylate in increasing population densities of PpG7 in soil also was affected by inoculum level, field site, and soil depth. Proportionate differences were greater in soils infested with approximately 10 CFU of PpG7 per g than in comparable soils infested with 10 CFU/g. In low-inoculum soils, increases from salicylate amendments were 26- and 29-fold in rhizosphere and nonrhizosphere soils, respectively, and in high-inoculum soils, the respective increases were 5.6- and 5-fold. No increases of fungi able to utilize salicylate were detected in soils amended with salicylate. However, soil fumigation with metham-sodium significantly reduced (P < 0.01) population densities of fungal salicylate utilizers in rhizosphere and nonrhizosphere soils. PMID- 16348985 TI - Enhanced growth and activity of a biocontrol bacterium genetically engineered to utilize salicylate. AB - Plasmid NAH7 was transferred from Pseudomonas putida PpG7 to P. putida R20 [R20(NAH7)], an antagonist of Pythium ultimum. The plasmid did not affect growth or survival of R20(NAH7) and was stably maintained under nonselective conditions in broth and soil and on sugar beet seeds. Plasmid NAH7 conferred to R20(NAH7) the ability to utilize salicylate in culture, agricultural field soil, and on sugar beet seeds. The metabolic activity of R20(NAH7), but not the wild-type R20, was greatly increased in soil by amendment with salicylate (250 mug/g) as measured by induced respiration. Population densities of R20(NAH7) were also enhanced in salicylate-amended soil, increasing from approximately 1 x 10 CFU/g to approximately 3 x 10 CFU/g after 35 h of incubation. In contrast, population densities of R20(NAH7) in nonamended soil were approximately 3 x 10 CFU/g of soil after 35 h of incubation. The concentration of salicylate in soil affected the rate and extent of population increase by R20(NAH7). At 50 to 250 mug of salicylate per g of soil, population densities of R20(NAH7) increased to approximately 10 CFU/g of soil by 48 h of incubation, with the fastest increase at 100 mug/g. A lag phase of approximately 24 h occurred before the population density increased in the presence of salicylate at 500 mug/g; at 1,000 mug/g, population densities of R20(NAH7) declined over the time period of the experiment. Population densities of R20(NAH7) on sugar beet seeds in soils amended with 100 mug of salicylate per g were not increased while ample carbon was present in the spermosphere. However, after carbon from the seed had been utilized, population densities of R20(NAH7) decreased significantly less (P = 0.005) on sugar beet seeds in soil amended with salicylate than in nonamended soil. PMID- 16348986 TI - Transformation of Kluyveromyces lactis by Electroporation. AB - The physical and biological parameters involved in efficient transformation of Kluyveromyces lactis by electroporation have been analyzed. By using an optimum voltage and a constant volume of cell suspension in a cuvette, the efficiency of transformation increased with increases in cell numbers and plasmid concentration. However, the most important parameter was the time of the pulse. Changes of 1 ms decreased the efficiency of transformation more than 70 to 80%. Under our best conditions, between 10 and 10 transformants per mug of plasmid DNA could be obtained. Under certain conditions, the size of the plasmid also affected electroporation efficiency. In any case, we did not obtain integrative transformation with an autonomously replicating plasmid. PMID- 16348987 TI - Nitrification and denitrification in lake and estuarine sediments measured by the N dilution technique and isotope pairing. AB - The transformation of nitrogen compounds in lake and estuarine sediments incubated in the dark was analyzed in a continuous-flowthrough system. The inflowing water contained NO(3), and by determination of the isotopic composition of the N(2), NO(3), and NH(4) pools in the outflowing water, it was possible to quantify the following reactions: total NO(3) uptake, denitrification based on NO(3) from the overlying water, nitrification, coupled nitrification denitrification, and N mineralization. In sediment cores from both lake and estuarine environments, benthic microphytes assimilated NO(3) and NH(4) for a period of 25 to 60 h after darkening. Under steady-state conditions in the dark, denitrification of NO(3) originating from the overlying water accounted for 91 to 171 mumol m h in the lake sediments and for 131 to 182 mumol m h in the estuarine sediments, corresponding to approximately 100% of the total NO(3) uptake for both sediments. It seems that high NO(3) uptake by benthic microphytes in the initial dark period may have been misinterpreted in earlier investigations as dissimilatory reduction to ammonium. The rates of coupled nitrification denitrification within the sediments contributed to 10% of the total denitrification at steady state in the dark, and total nitrification was only twice as high as the coupled process. PMID- 16348988 TI - Effects of Grazing by Flagellates on Competition for Ammonium between Nitrifying and Heterotrophic Bacteria in Soil Columns. AB - The enhanced mineralization of immobilized nitrogen by bacteriophagous protozoa has been thought to favor the nitrification process in soils in which nitrifying bacteria must compete with heterotrophic bacteria for the available ammonium. To obtain more insight into this process, the influence of grazing by the flagellate Adriamonas peritocrescens on the competition for ammonium between the chemolithotrophic species Nitrosomonas europaea and the heterotrophic species Arthrobacter globiformis in the presence of Nitrobacter winogradskyi was studied in soil columns, which were continuously percolated with media containing 5 mM ammonium and different amounts of glucose at a dilution rate of 0.007 h (liquid volumes). A. globiformis won the competition for ammonium. The grazing activities of the flagellates had two prominent effects on the competition between N. europaea and A. globiformis. First, the distribution of ammonium over the profile of the soil columns was more uniform in the presence of flagellates than in their absence. In the absence of flagellates, relatively high amounts of ammonium accumulated in the upper layer (0 to 3 cm), whereas in the underlying layers the ammonium concentrations were low. In the presence of flagellates, however, considerable amounts of ammonium were found in the lower layers, whereas less ammonium accumulated in the upper layer. Second, the potential ammonium-oxidizing activity of N. europaea was stimulated in the presence of flagellates. The numbers of N. europaea at different glucose concentrations in the presence of flagellates were comparable to those in the absence of protozoa. However, in the presence of flagellates, the potential ammonium-oxidizing activities were four to five times greater than those in the absence of protozoa. PMID- 16348989 TI - Purification and Characterization of Carbaryl Hydrolase from Blastobacter sp. Strain M501. AB - A bacterium capable of hydrolyzing carbaryl (1-naphthyl-N-methylcarbamate) was isolated from a soil enrichment. This bacterium was characterized taxonomically as a Blastobacter sp. and designated strain M501. A carbaryl hydrolase present in this strain was purified to homogeneity by protamine sulfate treatment, ammonium sulfate precipitation, and hydrophobic, anion-exchange, gel filtration, and hydroxylapatite chromatographies. The native enzyme had a molecular mass of 166,000 Da and was composed of two subunits with molecular masses of 84,000 Da. The optimum pH and temperature of the enzyme activity were 9.0 and 45 degrees C, respectively. The enzyme was not stable at temperatures above 40 degrees C. The purified enzyme hydrolyzed seven N-methylcarbamate insecticides and also exhibited activity against 1-naphthyl acetate and 4-nitrophenyl acetate. PMID- 16348990 TI - Effects of Condensed Tannins on Endoglucanase Activity and Filter Paper Digestion by Fibrobacter succinogenes S85. AB - The effect of condensed tannins from birdsfoot trefoil (Lotus corniculatus L.) on the cellulolytic rumen bacterium Fibrobacter succinogenes S85 was examined. Condensed tannins inhibited endoglucanase activity in the extracellular culture fluid, at concentrations as low as 25 mug ml. In contrast, cell-associated endoglucanase activity increased in concentrations of condensed tannins between 100 and 300 mug ml. Inhibition of endoglucanase activity in both the extracellular and the cell-associated fractions was virtually complete at 400 mug of condensed tannins ml. Despite the sharp decline in extracellular endoglucanase activity with increasing concentrations of condensed tannins, filter paper digestion declined only moderately between 0 and 200 mug of condensed tannins ml. However, at 300 mug ml, filter paper digestion was dramatically reduced and at 400 mug ml, almost no filter paper was digested. F. succinogenes S85 was seen to form digestive grooves on the surface of cellulose, and at 200 mug ml, digestive pits were formed which penetrated into the interior of cellulose fibers. Cells grown with condensed tannins (100 to 300 mug ml) possessed large amounts of surface material, and although this material may have been capsular carbohydrate, its osmiophilic nature suggested that it had arisen from the formation of tannin protein complexes on the cell surface. The presence of electron-dense extracellular material suggested that similar complexes were formed with extracellular protein. PMID- 16348991 TI - Enantiomeric Composition of the trans-Dihydrodiols Produced from Phenanthrene by Fungi. AB - The trans-dihydrodiols produced during the metabolism of phenanthrene by Cunninghamella elegans, Syncephalastrum racemosum, and Phanerochaete chrysosporium were purified by high-performance liquid chromatography (HPLC). The enantiomeric compositions and optical purities of the trans-dihydrodiols were determined to compare interspecific differences in the regio- and stereoselectivity of the fungal enzymes. Circular dichroism spectra of the trans dihydrodiols were obtained, and the enantiomeric composition of each preparation was analyzed by HPLC with a chiral stationary-phase column. The phenanthrene trans-1,2-dihydrodiol produced by C. elegans was a mixture of the 1R,2R and 1S,2S enantiomers in variable proportions. The phenanthrene trans-3,4-dihydrodiol produced by P. chrysosporium was the optically pure 3R,4R enantiomer, but that produced by S. racemosum was a 68:32 mixture of the 3R,4R and 3S,4S enantiomers. The phenanthrene trans-9,10-dihydrodiol produced by P. chrysosporium was predominantly the 9S,10S enantiomer, but those produced by C. elegans and S. racemosum were predominantly the 9R,10R enantiomer. The results indicate that although different fungi may exhibit similar regioselectivity, there still may be differences in stereoselectivity that depend on the species and the cultural conditions. PMID- 16348992 TI - Isolation of Typical Marine Bacteria by Dilution Culture: Growth, Maintenance, and Characteristics of Isolates under Laboratory Conditions. AB - Marine bacteria in Resurrection Bay near Seward, Alaska, and in the central North Sea off the Dutch coast were cultured in filtered autoclaved seawater following dilution to extinction. The populations present before dilution varied from 0.11 x 10 to 1.07 x 10 cells per liter. The mean cell volume varied between 0.042 and 0.074 mum, and the mean apparent DNA content of the cells ranged from 2.5 to 4.7 fg of DNA per cell. All three parameters were determined by high-resolution flow cytometry. All 37 strains that were obtained from very high dilutions of Resurrection Bay and North Sea samples represented facultatively oligotrophic bacteria. However, 15 of these isolates were eventually obtained from dilution cultures that could initially be cultured only on very low-nutrient media and that could initially not form visible colonies on any of the agar media tested, indicating that these cultures contained obligately oligotrophic bacteria. It was concluded that the cells in these 15 dilution cultures had adapted to growth under laboratory conditions after several months of nutrient deprivation prior to isolation. From the North Sea experiment, it was concluded that the contribution of facultative oligotrophs and eutrophs to the total population was less than 1% and that while more than half of the population behaved as obligately oligotrophic bacteria upon first cultivation in the dilution culture media, around 50% could not be cultured at all. During one of the Resurrection Bay experiments, 53% of the dilution cultures obtained from samples diluted more than 2.5 x 10 times consisted of such obligate oligotrophs. These cultures invariably harbored a small rod-shaped bacterium with a mean cell volume of 0.05 to 0.06 mum and an apparent DNA content of 1 to 1.5 fg per cell. This cell type had the dimensions of ultramicrobacteria. Isolates of these ultramicrobacterial cultures that were eventually obtained on relatively high-nutrient agar plates were, with respect to cell volume and apparent DNA content, identical to the cells in the initially obligately oligotrophic bacterial dilution culture. Determination of kinetic parameters from one of these small rod-shaped strains revealed a high specific affinity for the uptake of mixed amino acids (a degrees (A), 1,860 liters/g of cells per h), but not for glucose or alanine as the sole source of carbon and energy (a degrees (A), +/- 200 liters/g of cells per h). The ultramicrobial strains obtained are potentially a very important part of picoplankton biomass in the areas investigated. PMID- 16348993 TI - Nutrient Requirements of Renibacterium salmoninarum on Agar and in Broth Media. AB - In well-aerated broth cultures, good growth of Renibacterium salmoninarum was obtained in a serum-free medium consisting of 1% peptone, 1% yeast extract, and 0.1% l-cysteine (PYC broth). In contrast, serum or charcoal is required for growth on agar medium. Charcoal treatment of broth media, either before bacterial inoculation or during growth, increased the growth of R. salmoninarum, whereas the surfactants Tween 20 and Tween 80 inhibited growth. l-Cysteine was essential for optimal growth. Other organic sulfur compounds, such as d-cysteine, l methionine, homocysteine, homocysteine thiolactone, and reduced glutathione, supported only lower levels of growth, while cystine and dithiothreitol did not allow growth. PMID- 16348994 TI - Foliar Chlorosis in Symbiotic Host and Nonhost Plants Induced by Rhizobium tropici Type B Strains. AB - Rhizobium tropici CIAT899 induced chlorosis in the leaves of its symbiotic hosts, common bean (Phaseolus vulgaris L.), siratro (Macroptilium atropurpureum Urb.), and Leucaena leucocephala (Lam.) de Wit. Chlorosis induction by strains CIAT899 and CT9005, an exopolysaccharide-deficient mutant of CIAT899, required carbon substrate. When the bacteria were added at planting in a solution of mannitol (50 g/liter), as few as 10 cells of CIAT899 were sufficient to induce chlorosis in bean plants. All carbon sources tested, including organic acids and mono- and disaccharides, supported chlorosis induction. The addition of a carbon source did not affect the growth rate or the population density of CT9005 in the bean plant rhizosphere. Cell-free filtrates of cultures of CT9005 did not induce detectable chlorosis. All type B strains of R. tropici tested also induced chlorosis in common bean. Type A strains of R. tropici and all other species of bacteria tested did not induce chlorosis. Several lines of evidence indicated that nodulation was not required for chlorosis induction. Strain RSP900, a pSym-cured derivative of CIAT899, induced chlorosis in wild-type P. vulgaris. In addition, NOD125, a nodulation-defective line of common bean, developed chlorosis when inoculated with CIAT899, but did not develop nodules. CIAT899 consistently induced severe chlorosis in the leaves of the nonhost legumes alfalfa (Medicago sativa L.) and Berseem clover (Trifolium alexandrinum L.), and induced chlorosis in 29 to 58% of the plants tested of sunflower, cucumber, and tomato seedlings, but it did not induce chlorosis in the leaves of corn or wheat. Chlorosis induction in nonhost plants also required carbon substrate. The data are consistent with the hypothesis that R. tropici type B strains produce a chlorosis inducing factor that affects a wide range of plant species. PMID- 16348995 TI - Genetic Variation within a Lotic Population of Janthinobacterium lividum. AB - An understanding of the genetic variation within and between populations should allow scientists to address many problems, including those associated with endangered species and the release of genetically modified organisms into the environment. With respect to microorganisms, the release of genetically engineered microorganisms is likely to increase dramatically given the current growth in the bioremediation industry. In this study, genetic variation within a lotic, bacterial population of Janthinobacterium lividum was measured with restriction fragment length polymorphism analysis. Chromosomal DNA from 10 Kettle Creek (Hawk Mountain Sanctuary, Kempton, Pa.) J. lividum isolates was digested with six restriction endonucleases and probed with a 7.5-kb pKK3535 fragment containing the E. coli rrnB rRNA operon. Genetic variation, as measured in terms of nucleotide diversity, was high within the population. The 0.0781 value for genetic variation was especially high given the conservative nature of the genetic probe. The average percent similarity among isolates within the population was 67.25%. Pairwise comparisons of nucleotide diversity values (pi) and similarity coefficients (F) yielded values ranging from 0.0032 to 0.1816 and 0.3363 to 0.9808, respectively. Putative clonemates were not present within the group of isolates; however, all isolates shared 14 fragments across a spectrum of six restriction enzymes. The presence of these common fragments indicates that restriction fragment length polymorphism analysis may provide population- or species-specific diagnostic markers for J. lividum. Data that suggest a plume effect with respect to the downstream movement of J. lividum are also presented. An increase in genetic variation within groups of isolates along the longitudinal gradient of Kettle Creek is also suggested. PMID- 16348996 TI - Use of Fluorinated Compounds To Detect Aromatic Metabolites from m-Cresol in a Methanogenic Consortium: Evidence for a Demethylation Reaction. AB - Anaerobic sewage sludge was used to enrich a methanogenic m-cresol-degrading consortium. 6-Fluoro-3-methylphenol was synthesized and added to subcultures of the consortium with m-cresol. This caused the accumulation of 4-hydroxy-2 methylbenzoic acid. In a separate experiment, the addition of 3-fluorobenzoic acid caused the transient accumulation of 4-hydroxybenzoic acid. Inhibition with bromoethanesulfonic acid caused the accumulation of benzoic acid. Thus, the proposed degradation pathway was m-cresol --> 4-hydroxy-2-methylbenzoic acid --> 4-hydroxybenzoic acid --> benzoic acid. The m-cresol-degrading consortium was able to convert exogenous 4-hydroxybenzoic acid and benzoic acid to methane. In addition, for each metabolite of m-cresol identified, the corresponding fluorinated metabolite was detected, giving the following sequence: 6-fluoro-3 methylphenol --> 5-fluoro-4-hydroxy-2-methylbenzoic acid --> 3-fluoro-4 hydroxybenzoic acid --> 3-fluorobenzoic acid. The second step in each of these pathways is a novel demethylation which was rate limiting. This demethylation reaction would likely facilitate the transformation of the methyl group to methane, which is consistent with the results of a previous study that showed that the methyl carbon of m-[methyl-C]cresol was recovered predominantly as [C]methane (D. J. Roberts, P. M. Fedorak, and S. E. Hrudey, Can. J. Microbiol. 33:335-338, 1987). The final aromatic compound in the proposed route for m-cresol metabolism was benzoic acid, and its detection in these cultures merges the pathway for the methanogenic degradation of m-cresol with those for the anaerobic metabolism of many phenols. PMID- 16348997 TI - Reductive dechlorination of the nitrogen heterocyclic herbicide picloram. AB - The anaerobic biodegradation of picloram (3,5,6-trichloro-4-amino-2 pyridinecarboxylic acid) in freshwater sediment was favored under methanogenic conditions but not when sulfate or nitrate was available as a terminal electron acceptor. Under the former conditions, more than 85% of the parent substrate (340 muM) was removed from nonsterile incubations in 30 days, following a 50-day acclimation period. Concomitant with substrate decay, an intermediate transiently accumulated in the sediment slurries. By liquid chromatography-mass spectrometry, the intermediate was identified as an isomer of dichloro-4-amino-2 pyridinecarboxylic acid. Proton nuclear magnetic resonance evidence suggested that a chlorine was reductively removed from the parent substrate at the position meta to the nitrogen heteroatom. Upon continued incubation, the dechlorinated product was transformed into an unidentified compound which accumulated and resisted further decay. The addition of sulfate or bromoethanesulfonic acid to sediment slurries inhibited picloram dehalogenation, but molybdate reversed the inhibitory effect of sulfate on pesticide metabolism. These findings help clarify the fate of a halogenated nitrogen heterocyclic herbicide in anaerobic environments. PMID- 16348998 TI - Mobilization of a Recombinant IncQ Plasmid between Bacteria on Agar and in Soil via Cotransfer or Retrotransfer. AB - Mobilization of a genetically engineered IncQ plasmid, pSKTG, was studied in vitro and in sterile and nonsterile soils. In biparental and triparental filter matings, the mobilization frequencies of pSKTG were identical, and the plasmid was mobilized only in the presence of self-transmissible plasmid RP4p. In sterile soil, mobilization was probably limited by reduced cell-to-cell contact, since the frequencies of mobilization were approximately 100-fold lower than the frequencies in the filter matings. The transfer frequency of pSKTG in sterile soil when RP4p was present in the same strain was about 100-fold higher than the transfer frequency when RP4p was present in a separate strain. In studies in natural soil, pSKTG was also found to be transferred to indigenous bacteria. However, natural mobilization by genetic elements present in the indigenous soil microflora could not be detected. In vitro studies of natural transfer suggested that such genetic elements occur in soil bacteria. PMID- 16348999 TI - C(15)H(24) Volatile Compounds Unique to Aflatoxigenic Strains of Aspergillus flavus. AB - Headspace volatiles from eight strains of Aspergillus flavus (four aflatoxigenic strains and four nonaflatoxigenic strains), grown for 1, 2, 3, 4, 8, and 10 days in submerged cultures, were collected in Tenax GC traps. The traps were desorbed onto a 50-m gas-liquid chromatography capillary column by heat and gas purge from an external direct injector device. The column was interfaced with a mass spectrometer data acquisition system. Peaks were identified by comparing retention times and mass spectra with those obtained from authentic compounds and by using a computer-assisted mass spectral data base. Aflatoxigenic strains of A. flavus produced several C(15)H(24) compounds (e.g., alpha-gurjunene, trans caryophyllene, and cadinene) which peaked in 3-day cultures and were not present in earlier (1- and 2-day) or later (8- and 10-day) cultures. None of these volatiles were detected in nonaflatoxigenic strains of A. flavus. There was an apparent correlation between the release of C(15)H(24) volatile compounds and the initiation of aflatoxin biosynthesis, and a correlation between decline of aflatoxin synthesis and the disappearance of the C(15)H(24) compounds unique to aflatoxigenic A. flavus also existed. PMID- 16349000 TI - Effects of Grazing by the Free-Living Soil Amoebae Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis on Various Bacteria. AB - Cultures of 10 different bacteria were used to serve as food sources for axenically grown Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. The nonpigmented enterobacteriaceae Escherichia coli K 12 and Klebsiella aerogenes appeared to be excellent feed to all three amoebae. Hardly any growth or ammonium production was observed in tests with Chromatium vinosum and Serratia marcescens, which share the presence of pigmented compounds. Distinct differences in net ammonium production were detected and were correlated to the amoebal growth yield. In general, growth of amoebae and ammonium production increased in the order A. polyphaga, A. castellanii, and H. vermiformis. PMID- 16349001 TI - Cell Density and Growth Phase as Factors in the Resistance of a Biofilm of Pseudomonas aeruginosa (ATCC 27853) to Iodine. AB - Previous studies have shown that biofilms exhibit enhanced resistance to iodine. Investigations were conducted to determine the relative importance of growth phase versus cell density on biofilm resistance of Pseudomonas aeruginosa (ATCC 27853) to iodine. Cell density is a contributing factor to resistance, whereas growth to the stationary phase is not sufficient to achieve resistance. PMID- 16349002 TI - Two widely accessible media for growth and reproduction of phytophthora and pythium species. AB - Tomato agar and soybean agar were found to be comparable to or in some cases better than the popular V8 vegetable juice agar in supporting linear growth of Phytophthora cactorum, Phytophthora capsici, Phytophthora parasitica, Pythium aphanidermatum, and Pythium splendens; sporangium production of P. capsici, P. palmivora, and Pythium splendens; and oospore formation of P. cactorum, P. parasitica, Pythium aphanidermatum and Pythium splendens. These two media with readily accessible ingredients can be used to substitute for V8 juice agar in pathological and physiological studies of these two important groups of fungi in countries where V8 vegetable juice is not available or difficult to obtain. PMID- 16349004 TI - Sorption of heavy metals to the filamentous bacterium thiothrix strain A1. AB - [This corrects the article on p. 1278 in vol. 59.]. PMID- 16349003 TI - Glutamate Dehydrogenase Is Not Essential for Glutamate Formation by Corynebacterium glutamicum. AB - Two Corynebacterium glutamicum strains, one being glutamate dehydrogenase (GDH) negative and the other possessing 11-fold-higher specific GDH activity than the parental wild type, were constructed and used to analyze the role of GDH in C. glutamicum. The results indicate (i) that GDH is dispensable for glutamate synthesis required for growth and (ii) that although a high level of GDH increases the intracellular glutamate pool, the level of GDH has no influence on glutamate secretion. PMID- 16349005 TI - Localization and Characterization of alpha-Glucosidase Activity in Brettanomyces lambicus. AB - Brettanomyces lambicus was isolated and identified from a typical overattenuating Belgian lambic beer and exhibited extracellular and intracellular alpha glucosidase activities. Production of the intracellular enzyme was higher than production of the extracellular enzyme, and localization studies showed that the intracellular alpha-glucosidase is mostly soluble and partially cell wall bound. Both intracellular and extracellular enzymes were purified by ammonium sulfate precipitation, gel filtration (Sephadex G-150, Sephadex G-200, Ultrogel AcA-44), and ion-exchange chromatography (sulfopropyl-Sephadex C-50, (carboxymethyl Sephadex C-50). The intracellular alpha-glucosidase exhibited optimum activity at 39 degrees C and pH 6.2. The extracellular enzyme exhibited optimum catalytic activity at 40 degrees C and pH 6.0. The molecular masses of purified intracellular and extracellular alpha-glucosidases, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were 72,500 and 77,250, respectively. For both enzymes there was a decrease in the rate of hydrolysis with an increase in the degree of polymerization, and both enzymes hydrolyzed dextrins isolated from lambic wort (degrees of polymerization, 3 to 9 and more than 9). The K(m) values for p-nitrophenyl-alpha-d-glucopyranoside, maltose, and maltotriose for the intracellular enzyme were 0.9, 3.4, and 3.7 mM, respectively. The K(i) values for both enzymes were between 28.5 and 57 muM for acarbose and between 7.45 and 15.7 mM for Tris. These enzymes are probably involved in the overattenuation of spontaneously fermented lambic beer. PMID- 16349006 TI - Coal Depyritization by the Thermophilic Archaeon Metallosphaera sedula. AB - The kinetics of pyrite oxidation by Metallosphaera sedula were investigated with mineral pyrite and two coals with moderate (Pittsburgh no. 8) and high (New Brunswick, Canada) pyritic sulfur content. M. sedula oxidized mineral pyrite at a greater rate than did another thermophile, Acidianus brierleyi, or a mesophile, Thiobacillus ferrooxidans. Maximum rates of coal depyritization were also greater with M. sedula, although the magnitude of biological stimulation above abiotic rates was notably less than with mineral pyrite. Coal depyritization appears to be limited by the oxidation of pyrite with ferric ions and not by the rate of biotic oxidation of ferrous iron, as evidenced by the maintenance of a high ratio of ferric to ferrous iron in solution by M. sedula. Significant precipitation of hydronium jarosite at elevated temperature occurred only with New Brunswick coal. PMID- 16349007 TI - Development of steady-state diffusion gradients for the cultivation of degradative microbial consortia. AB - A diffusion gradient plate was constructed and evaluated for its potential use in the isolation of degradative microbial consortia from natural habitats. In this model, a steady-state concentration gradient of diclofop methyl, established by diffusion through an agarose gel, provided the carbon for microbial growth. Colonization of the gel surface was observed with epifluorescence and scanning confocal laser microscopy to determine microbial responses to the diclofop gradient. A detectable gradient developed over a narrow band (<10 mm). Consequently, quantitative analyses of the microbial response to the gradient were difficult to obtain. A two-dimensional, finite-element numerical transport model for advective-diffusive transport was used to simulate concentration and flux profiles in the physical model. The simulated profiles were correlated with the measured concentration gradient (R = 0.89) and the cell numbers on the gel surface (R = 0.85). The numerical model was subsequently used to redesign the physical model. The detectable concentration gradient in the modified physical model extended over the length of the gel (38 mm). The simulated profile again showed a good correlation with the measured profile (R = 0.96) and the microbial responses to the concentration gradient (R = 0.99). It was concluded that these gradients provide the steady-state environments needed to sustain steady-state consortia. They also provide a physical pathway for the development of degradative biofilms from low to high concentrations of toxicants and simulate conditions under which low concentrations of toxicant are supplied at a constant flux over long periods of time, such as the conditions that could occur in natural environments. PMID- 16349008 TI - Dominating role of an unusual magnetotactic bacterium in the microaerobic zone of a freshwater sediment. AB - A combination of polymerase chain reaction-assisted rRNA sequence retrieval and fluorescent oligonucleotide probing was used to identify in situ a hitherto unculturable, big, magnetotactic, rod-shaped organism in freshwater sediment samples collected from Lake Chiemsee. Tentatively named "Magnetobacterium bavaricum," this bacterium is evolutionarily distant from all other phylogenetically characterized magnetotactic bacteria and contains unusually high numbers of magnetosomes (up to 1,000 magnetosomes per cell). The spatial distribution in the sediment was studied, and up to 7 x 10 active cells per cm were found in the microaerobic zone. Considering its average volume (25.8 +/- 4.1 mum) and relative abundance (0.64 +/- 0.17%), "M. bavaricum" may account for approximately 30% of the microbial biovolume and may therefore be a dominant fraction of the microbial community in this layer. Its microhabitat and its high content of sulfur globules and magnetosomes suggest that this organism has an iron-dependent way of energy conservation which depends on balanced gradients of oxygen and sulfide. PMID- 16349009 TI - Measurement of rRNA Variations in Natural Communities of Microorganisms on the Southeastern U.S. Continental Shelf. AB - The development of a clear understanding of the physiology of marine prokaryotes is complicated by the difficulties inherent in resolving the activity of various components of natural microbial communities. Application of appropriate molecular biological techniques offers a means of overcoming some of these problems. In this regard, we have used direct probing of bulk RNA purified from selective size fractions to examine variations in the rRNA content of heterotrophic communities and Synechococcus populations on the southeastern U.S. continental shelf. Heterotrophic communities in natural seawater cultures amended with selected substrates were examined. Synechococcus populations were isolated from the water column by differential filtration. The total cellular rRNA content of the target populations was assayed by probing RNA purified from these samples with an oligonucleotide complementing a universally conserved region in the eubacterial 16S rRNA (heterotrophs) or with a 1.5-kbp fragment encoding the Synechococcus sp. strain WH 7803 16S rRNA (cyanobacteria). The analyses revealed that heterotrophic bacteria responded to the addition of glucose and trace nutrients after a 6-h lag period. However, no response was detected after amino acids were added. The cellular rRNA content increased 48-fold before dropping to a value 20 times that detected before nutrients were added. Variations in the rRNA content from Synechococcus spp. followed a distinct diel pattern imposed by the phasing of cell division within the irradiance cycle. The results indicate that careful application of these appropriate molecular biological techniques can be of great use in discerning basic physiological characteristics of selected natural populations and the mechanisms which regulate growth at the subcellular level. PMID- 16349010 TI - Effect of Increasing the Copy Number of Bacteriophage Origins of Replication, in trans, on Incoming-Phage Proliferation. AB - Bacteriophage resistance mechanisms which are derived from a bacteriophage genome are termed Per (phage-encoded resistance). When present in trans in Lactococcus lactis NCK203, Per50, the cloned origin of replication from phage phi50, interferes with phi50 replication. The per50 fragment was found to afford negligible protection to NCK203 against phi50 infection when present in a low copy-number plasmid, pTRK325. A high-copy-number Per50 construct (pTRK323) dramatically affected phi50 infection, reducing the efficiency of plaquing (EOP) to 2.5 x 10 and the plaque size to pinhead proportions. This clone also afforded significant protection against other related small isometric phages. Per31 was cloned from phage phi31 and demonstrated to function as an origin of replication by enabling replication of per31-containing plasmids, in NCK203, on phi31 infection. A low-copy-number Per31 plasmid (pTRK360) reduced the EOP of phi31 on NCK203 to 0.3 and the plaque diameter from 1.5 to 0.5 mm. When this plasmid was cloned in high copy number, the EOP was further reduced to 7.2 x 10 but the plaques were large and contained Per31-resistant phages. Characterization of these "new" phages revealed at least two different types that were similar to phi31, except that DNA alterations were noted in the region containing the origin. This novel and powerful abortive phage resistance mechanism should prove useful when directed at specific, problematic phages. PMID- 16349011 TI - Selective inhibition of ammonium oxidation and nitrification-linked n(2)o formation by methyl fluoride and dimethyl ether. AB - Methyl fluoride (CH(3)F) and dimethyl ether (DME) inhibited nitrification in washed-cell suspensions of Nitrosomonas europaea and in a variety of oxygenated soils and sediments. Headspace additions of CH(3)F (10% [vol/vol]) and DME (25% [vol/vol]) fully inhibited NO(2) and N(2)O production from NH(4) in incubations of N. europaea, while lower concentrations of these gases resulted in partial inhibition. Oxidation of hydroxylamine (NH(2)OH) by N. europaea and oxidation of NO(2) by a Nitrobacter sp. were unaffected by CH(3)F or DME. In nitrifying soils, CH(3)F and DME inhibited N(2)O production. In field experiments with surface flux chambers and intact cores, CH(3)F reduced the release of N(2)O from soils to the atmosphere by 20- to 30-fold. Inhibition by CH(3)F also resulted in decreased NO(3) + NO(2) levels and increased NH(4) levels in soils. CH(3)F did not affect patterns of dissimilatory nitrate reduction to ammonia in cell suspensions of a nitrate-respiring bacterium, nor did it affect N(2)O metabolism in denitrifying soils. CH(3)F and DME will be useful in discriminating N(2)O production via nitrification and denitrification when both processes occur and in decoupling these processes by blocking NO(2) and NO(3) production. PMID- 16349012 TI - pH and Glucose Profiles in Aggregates of Bacillus laevolacticus. AB - Size distributions and glucose and pH profiles of aggregates of the d-(-)-lactic acid-producing organism Bacillus laevolacticus were measured. The organisms were grown in continuous culture with a medium glucose concentration of either 280 or 110 mM. A maximal aggregate diameter of 2.2 mm, with a Sauter mean of 1.46 mm, was determined for the former culture condition, whereas aggregates from a culture with 110 mM glucose input had a maximal diameter of 1.9 mm (Sauter mean of 1.07 mm). A pH gradient of approximately 2 U was observed for large aggregates (above 1.5 mm). In smaller aggregates (0.75 mm), the pH value in the interior part was approximately 0.4 U lower than that in the culture fluid. It could be concluded that, in cultures with the high glucose input, lactic acid accumulated within the aggregates to such an extent that metabolism in the central region of the larger aggregates could not proceed further. In these cultures, approximately 90% of the total biomass was active. In aggregates from cultures with a low glucose input, glucose only partly penetrated the larger-sized aggregates, and the activity of this culture was reduced to approximately 70% of the biomass. These aggregates were found to decrease in size after prolonged periods of cultivation. It is suggested that this is caused by glucose depletion in the interior of the aggregates. It is concluded that the availability of glucose is an important factor in determining the size of aggregates of B. laevolacticus. PMID- 16349013 TI - Methylmercury Resistance in Desulfovibrio desulfuricans Strains in Relation to Methylmercury Degradation. AB - Two strains of Desulfovibrio desulfuricans, one known to synthesize monomethylmercury from ionic mercury, were grown to determine methylmercury toxicity and for comparison with an anaerobic strain of Clostridium pasteurianum, a H(2) producer, and with the broad-spectrum mercury-resistant Pseudomonas putida strain FB-1, capable of degrading 1 mug of methylmercury to methane and elemental mercury in 2 h. The CH(3)HgCl resistance of D. desulfuricans strains was 10 times that of P. putida FB-1 and 100 times that of C. pasteurianum. The methylmercury resistance of D. desulfuricans was related to the disappearance of methylmercury from cultures by transformation to dimethylmercury, metacinnabar, methane, and traces of ionic mercury. During a 15-day experiment the kinetics of the two volatile compounds dimethylmercury [(CH(3))(2)Hg] and methane were monitored in the liquid by a specific new technique with purge-and-trap gas chromatography in line with Fourier transform infrared spectroscopy and in the headspace by gas chromatography with flame ionization detection. Insoluble metacinnabar (cubic HgS) of biological origin was detected by X-ray diffractometry in the gray precipitate from the insoluble residue of the pellet of a 1-liter culture spiked with 100 mg of CH(3)HgCl. This was compared with a 1-liter culture of D. desulfuricans LS spiked with 100 mg of HgCl(2). In a further experiment, it was demonstrated that insoluble, decomposable, white dimethylmercury sulfide [(CH(3)Hg)(2)S] formed instantly in the reaction of methylmercury with hydrogen sulfide. This organomercurial was extracted with chloroform and identified by gas chromatography in line with mass spectrometry. The D. desulfuricans strains were resistant to high concentrations of methylmercury because they produced insoluble dimethylmercury sulfide, which slowly decomposed under anaerobic conditions to metacinnabar and volatilized to dimethylmercury and methane between pHs 6.2 and 6.5 for high (4.5-g . liter) or low (0.09-g . liter) sulfate contents. Methane was produced from CH(3)HgCl at a lower rate than by the broad-spectrum Hg resistant P. putida strain FB-1. PMID- 16349014 TI - Kinetic Studies of Ammonia Monooxygenase Inhibition in Nitrosomonas europaea by Hydrocarbons and Halogenated Hydrocarbons in an Optimized Whole-Cell Assay. AB - The inhibitory effects of 15 hydrocarbons and halogenated hydrocarbons on NH(3) oxidation by ammonia monooxygenase (AMO) in intact cells of the nitrifying bacterium Nitrosomonas europaea were determined. Determination of AMO activity, measured as NO(2) production, required coupling of hydroxylamine oxidoreductase (HAO) activity with NH(3)-dependent NH(2)OH production by AMO. Hydrazine, an alternate substrate for HAO, was added to the reaction mixtures as a source of reductant for AMO. Most inhibitors exhibited competitive or noncompetitive inhibition patterns. The competitive character generally decreased (K(i) (E)/K(i) (ES) increased) as the molecular size of the inhibitors increased. For example, CH(4) and C(2)H(4) were competitive inhibitors of NH(3) oxidation, whereas the remaining alkanes (up to C(4)) and monohalogenated (Cl, Br, I) alkanes were noncompetitive. Oxidation of C(2)H(5)Br (noncompetitive) increased as the NH(4) concentration increased up to 40 mM, whereas oxidations of inhibitors with competitive character (K(i) (E) << K(i) (ES)) were diminished at 40 mM NH(4). Multichlorinated compounds produced nonlinear Lineweaver-Burk plots. Iodinated alkanes (CH(3)I, C(2)H(5)I) and C(2)Cl(4) were potent inhibitors of NH(3) oxidation. Maximum rates of NH(3), C(2)H(4), and C(2)H(6) oxidations were approximately equivalent, suggesting a common rate-determining step. These data support an active-site model for AMO consisting of an NH(3)-binding site and a second site that binds noncompetitive inhibitors, with oxidation occurring at either site. PMID- 16349015 TI - Stable Tagging of Rhizobium meliloti with the Firefly Luciferase Gene for Environmental Monitoring. AB - A system for stable tagging of gram-negative bacteria with the firefly luciferase gene, luc, is described. A previously constructed fusion constitutively expressing luc from the lambdap(R) promoter was used. Stable integration into the bacterial genome was achieved by use of mini-Tn5 delivery vectors. The procedure developed was applied for tagging of representative gram-negative bacteria, such as Escherichia coli, Rhizobium meliloti, Pseudomonas putida, and Agrobacterium tumefaciens. The system permitted the detection of tagged R. meliloti in the presence of more than 10 CFU per plate without the use of any selective markers (such as antibiotic resistance genes). No significant differences in growth rates or soil survival were found between the marked strain and the wild-type strain. Studies of bioluminescent R. meliloti also revealed a good correlation between cell biomass and bioluminescence. The firefly luciferase tagging system is an easy, safe, and sensitive method for the detection and enumeration of bacteria in the environment. PMID- 16349016 TI - Attachment of agrobacteria to grape cells. AB - The presence of the Ti plasmid favorably influences the attachment of agrobacteria to grape callus cells, especially during the early stages of a 2-h incubation. Agrobacterium strains attached to a similar extent to both the crown gall-resistant cultivar (Catawba), Vitis labruscana, and the crown gall susceptible cultivar (Chancellor), Vitis sp. Attachment of the virulent strain to grape callus cells is blocked by the avirulent strain HLB-2 in both the tissue culture cell suspension and the seedling root systems. PMID- 16349017 TI - A Cytochemical Study of Extracellular Sheaths Associated with Rigidoporus lignosus during Wood Decay. AB - An ultrastructural and cytochemical investigation of the development of Rigidoporus lignosus, a white-rot fungus inoculated into wood blocks, was carried out to gain better insight into the structure and role of the extracellular sheaths produced by this fungus during wood degradation. Fungal sheaths had a dense or loose fibrillar appearance and were differentiated from the fungal cell wall early after wood inoculation. Close association between extracellular fibrils and wood cell walls was observed at both early and advanced stages of wood alteration. Fungal sheaths were often seen deep in host cell walls, sometimes enclosing residual wood fragments. Specific gold probes were used to investigate the chemical nature of R. lignosus sheaths. While labeling of chitin, pectin, beta-1,4- and beta-1,3-glucans, beta-glucosides, galactosamine, mannose, sialic acid, RNA, fucose, and fimbrial proteins over fungal sheaths did not succeed, galactose residues and laccase (a fungal phenoloxidase) were found to be present. The positive reaction of sheaths with the PATAg test indicates that polysaccharides such as beta-1,6-glucans are important components. Our data suggest that extracellular sheaths produced by R. lignosus during host cell colonization play an important role in wood degradation. Transportation of lignin degrading enzymes by extracellular fibrils indicates that alteration of plant polymers may occur within fungal sheaths. It is also proposed that R. lignosus sheaths may be involved in recognition mechanisms in fungal cell-wood surface interactions. PMID- 16349018 TI - Estimating the growth rate of slowly growing marine bacteria from RNA content. AB - In past studies of enteric bacteria such as Escherichia coli, various measures of cellular RNA content have been shown to be strongly correlated with growth rate. We examined this correlation for four marine bacterial isolates. Isolates were grown in chemostats at four or five dilution rates, yielding growth rates that spanned the range typically determined for marine bacterial communities in nature (mu = 0.01 to 0.25 h). All measures of RNA content (RNA cell, RNA:biovolume ratio, RNA:DNA ratio, RNA:DNA:biovolume ratio) were significantly different among isolates. Normalizing RNA content to cell volume substantially reduced, but did not eliminate, these differences. On average, the correlation between mu and the RNA:DNA ratio accounted for 94% of variance when isolates were considered individually. For data pooled across isolates (analogous to an average measurement for a community), the ratio of RNA:DNA mum (cell volume) accounted for nearly half of variance in mu (r = 0.47). The maximum RNA:DNA ratio for each isolate was extrapolated from regressions. The regression of (RNA:DNA)/(RNA:DNA)(max) on mu was highly significant (r = 0.76 for data pooled across four isolates) and virtually identical for three of the four isolates, perhaps reflecting an underlying common relationship between RNA content and growth rate. The dissimilar isolate was the only one derived from sediment. Cellular RNA content is likely to be a useful predictor of growth rate for slowly growing marine bacteria but in practice may be most successful when applied at the level of individual species. PMID- 16349019 TI - Characterization of Amylolytic Enzymes, Having Both alpha-1,4 and alpha-1,6 Hydrolytic Activity, from the Thermophilic Archaea Pyrococcus furiosus and Thermococcus litoralis. AB - Extracellular pullulanases were purified from cell-free culture supernatants of the marine thermophilic archaea Thermococcus litoralis (optimal growth temperature, 90 degrees C) and Pyrococcus furiosus (optimal growth temperature, 98 degrees C). The molecular mass of the T. litoralis enzyme was estimated at 119,000 Da by electrophoresis, while the P. furiosus enzyme exhibited a molecular mass of 110,000 Da under the same conditions. Both enzymes tested positive for bound sugar by the periodic acid-Schiff technique and are therefore glycoproteins. The thermoactivity and thermostability of both enzymes were enhanced in the presence of 5 mM Ca, and under these conditions, enzyme activity could be measured at temperatures of up to 130 to 140 degrees C. The addition of Ca also affected substrate binding, as evidenced by a decrease in K(m) for both enzymes when assayed in the presence of this metal. Each of these enzymes was able to hydrolyze, in addition to the alpha-1,6 linkages in pullulan, alpha-1,4 linkages in amylose and soluble starch. Neither enzyme possessed activity against maltohexaose or other smaller alpha-1,4-linked oligosaccharides. The enzymes from T. litoralis and P. furiosus appear to represent highly thermostable amylopullulanases, versions of which have been isolated from less-thermophilic organisms. The identification of these enzymes further defines the saccharide metabolizing systems possessed by these two organisms. PMID- 16349020 TI - Starvation-induced thermal tolerance as a survival mechanism in a psychrophilic marine bacterium. AB - Carbon-starved cultures of strain Ant-300, a psychrophilic marine vibrio isolated from the Antarctic Convergence, were compared with their nonstarved counterparts for resistance to heat. Specifically, starved and unstarved cells were exposed to 17 degrees C, which is 4 degrees C above the maximum growth temperature, and compared with cells maintained at the optimum temperature (5 to 7 degrees C). Total cell counts, direct viable-cell counts, and plate counts were monitored. At a temperature of 17 degrees C, viability (as indicated by plate counts) was lost within 40 h, with direct viable-cell counts indicating less than 5% viability at this time. However, when cells were carbon starved for 1 week prior to heat challenge, significant plateability was maintained for more than 6 days; direct viable-cell counts of starved cells maintained at 17 degrees C indicated the presence of viable cells for at least 12 days. Because starvation is the normal physiological state of copiotrophic, heterotrophic bacteria in oligotrophic marine waters, these data suggest that starvation conditions may be a significant factor in providing heat tolerance to psychrophiles. PMID- 16349021 TI - Nonenzymatic Glycosylation of Lepidopteran-Active Bacillus thuringiensis Protein Crystals. AB - We used high-pH anion-exchange chromatography with pulsed amperometric detection to quantify the monosaccharides covalently attached to Bacillus thuringiensis HD 1 (Dipel) crystals. The crystals contained 0.54% sugars, including, in decreasing order of prevalence, glucose, fucose, arabinose/rhamnose, galactose, galactosamine, glucosamine, xylose, and mannose. Three lines of evidence indicated that these sugars arose from nonenzymatic glycosylation: (i) the sugars could not be removed by N- or O-glycanases; (ii) the sugars attached were influenced both by the medium in which the bacteria had been grown and by the time at which the crystals were harvested; and (iii) the chemical identity and stoichiometry of the sugars detected did not fit any known glycoprotein models. Thus, the sugars detected were the product of fermentation conditions rather than bacterial genetics. The implications of these findings are discussed in terms of crystal chemistry, fermentation technology, and the efficacy of B. thuringiensis as a microbial insecticide. PMID- 16349022 TI - Influence of Metronidazole, CO, CO(2), and Methanogens on the Fermentative Metabolism of the Anaerobic Fungus Neocallimastix sp. Strain L2. AB - The effects of metronidazole, CO, methanogens, and CO(2) on the fermentation of glucose by the anaerobic fungus Neocallimastix sp. strain L2 were investigated. Both metronidazole and CO caused a shift in the fermentation products from predominantly H(2), acetate, and formate to lactate as the major product and caused a lower glucose consumption rate and cell protein yield. An increased lactate dehydrogenase activity and a decreased hydrogenase activity were observed in cells grown under both culture conditions. In metronidazole-grown cells, the amount of hydrogenase protein was decreased compared with the amount in cells grown in the absence of metronidazole. When Neocallimastix sp. strain L2 was cocultured with the methanogenic bacterium Methanobrevibacter smithii, the fermentation pattern changed in the opposite direction: H(2) and acetate production increased at the expense of the electron sink products lactate, succinate, and ethanol. A concomitant decrease in the enzyme activities leading to these electron sink products was observed, as well as an increase in the glucose consumption rate and cell protein yield, compared with those of pure cultures of the fungus. Low levels of CO(2) in the gas phase resulted in increased H(2) and lactate formation and decreased production of formate, acetate, succinate, and ethanol, a decreased glucose consumption rate and cell protein yield, and a decrease in most of the hydrogenosomal enzyme activities. None of the tested culture conditions resulted in changed quantities of hydrogenosomal proteins. The results indicate that manipulation of the pattern of fermentation in Neocallimastix sp. strain L2 results in changes in enzyme activities but not in the proliferation or disappearance of hydrogenosomes. PMID- 16349023 TI - Identification of Vibrio splendidus as a Member of the Planktonic Luminous Bacteria from the Persian Gulf and Kuwait Region with luxA Probes. AB - Hybridization probes specific for the luxA genes of four groups of luminous bacteria were used to screen luminous isolates obtained from the Persian Gulf, near Al Khiran, Kuwait Nine of these isolates were identified as Vibrio harveyi, a commonly encountered planktonic isolate, while three others showed no hybridization to any of the four probes (V. harveyi, Vibrio fischeri, Photobacterium phosphoreum, or Photobacterium leiognathi) under high-stringency conditions. Polymerase chain reaction amplification was used to prepare a luxA probe against one of these isolates, K-1, and this probe was screened under high stringency conditions against a collection of DNAs from luminous bacteria; it was found to hybridize specifically to the DNA of the species Vibrio splendidus. A probe prepared against the type strain of V. splendidus (ATCC 33369) was tested against the collection of luminous bacterial DNA preparations and against the Kuwait isolates and was found to hybridize only against the type strain and the three unidentified Kuwait isolates. Extensive taxonomic analysis by standard methods confirmed the identification of the 13 isolates. PMID- 16349024 TI - Cloning of Protocatechuate 3,4-Dioxygenase Genes from Bradyrhizobium japonicum USDA110. AB - A heterologous gene probe encoding the alpha and beta subunits of the Pseudomonas cepacia protocatechuate 3,4-dioxygenase (PCD) was used to detect its homolog in the genome of Bradyrhizobium japonicum USDA110. Three cosmid clones carrying a 2.2-kb BamHI insert showed high levels of PCD activity. SacI digestion of one of the genomic clones, pBjG17, produced a 2.5-kb insert DNA that complemented a PCD mutant of P. cepacia. PMID- 16349025 TI - An Alkalophilic Bacillus sp. Produces 2-Phenylethylamine. AB - A large amount of 2-phenylethylamine was produced in cells of alkalophilic Bacillus sp. strain YN-2000. This amine is secreted in the medium during the cell growth. The amounts of 2-phenylethylamine in both cells and medium change upon changing the pH of the medium. PMID- 16349026 TI - Effects of methylated, organic, and inorganic substrates on microbial consumption of dimethyl sulfide in estuarine waters. AB - We examined the effects of a variety of amendments on the consumption of [U C]dimethyl sulfide in a Georgia salt marsh. Methylated compounds, particularly those with dimethyl groups, significantly inhibited dimethyl sulfide consumption, while nonmethylated substrates had little effect. Dimethyl disulfide and dimethyl ether were the most effective inhibitors tested. PMID- 16349027 TI - Composition of Non-Microbially Reducible Fe(III) in Aquatic Sediments. AB - The production of small quantities of Fe(II) during the initial phase of microbial Fe(III) reduction greatly increased the amount of Fe(III) that could be extracted from freshwater sediments with oxalate. This finding and other evidence suggest that the oxalate-extractable Fe(III) that is unavailable for microbial reduction in anoxic sediments is not in the form of mixed Fe(III)-Fe(II) forms, as was previously suggested, but rather is in the form of highly crystalline Fe(III) oxides. PMID- 16349028 TI - Rapid Mini-Prep Isolation of High-Quality Plasmid DNA from Lactococcus and Lactobacillus spp. AB - A simple, rapid plasmid mini-prep procedure for lactococci and lactobacilli which gives high yields and can be performed on overnight broth cultures is presented. Large plasmids were isolated from both lactococci and lactobacilli, including a 70-kb plasmid from Lactobacillus acidophilus C7. The purity of the resulting plasmid DNA makes it suitable for subsequent molecular manipulations. The convenience of the technique makes this rapid mini-prep procedure suitable for routine plasmid isolation from lactic acid bacteria. PMID- 16349029 TI - Recombination-Deficient Mutants of an Extreme Thermophile, Thermus thermophilus. AB - Recombination-deficient strains of the extreme thermophile Thermus thermophilus have been prepared from a leucine-isoleucine mutant strain (NM6). The availability of such recombination-deficient thermophilic bacterial strains may provide especially good hosts for work with plasmid vectors. PMID- 16349030 TI - Stimulation of vesicular-arbuscular mycorrhizal fungi by mycotrophic and nonmycotrophic plant root systems. AB - Transformed root cultures of three nonmycotrophic and one mycotrophic plant species stimulated germination and hyphal growth of the vesicular-arbuscular mycorrhizal fungus Glomus etunicatum (Becker & Gerd.) in a gel medium. However, only roots of the mycotrophic species (carrot) supported continued hyphal exploration after 3 to 4 weeks and promoted appressoria formation by G. etunicatum. PMID- 16349031 TI - Detection of mRNA in streptomyces cells by whole-cell hybridization with digoxigenin-labeled probes. AB - Detection of mRNA of the thiostrepton resistance gene (tsr) harbored by plasmid pIJ673 in Streptomyces violacelatus was achieved by whole-cell hybridization with digoxigenin-labeled in vitro transcripts followed by an antibody-alkaline phosphatase detection of the digoxigenin reporter molecule. Prior to hybridization, the cells had to be permeabilized by lysozyme, the detergent Nonidet P-40, and toluene. The permeability of the S. violacelatus cells for probes and the antibody-alkaline phosphatase conjugate was demonstrated by hybridization with digoxigenin-labeled, 16S rRNA-targeted oligonucleotides. PMID- 16349032 TI - Degradation of 2-Chloroethylvinylether by Ancylobacter aquaticus AD25 and AD27. AB - Incubation of five different beta-chloroethers with slurries prepared from brackish water sediment or activated sludge revealed that bis(2-chloroethyl)ether and 2-chloroethylvinylether (2-CVE) were biodegradable under aerobic conditions. After enrichment, two different cultures of Ancylobacter aquaticus that are capable of growth on 2-CVE were isolated. Both cultures were also able to grow on 1,2-dichloroethane. The cells contained a haloalkane dehalogenase that dehalogenated 2-CVE, 2-chloroethylmethylether, 2-bromoethylethylether, and epichlorohydrin. Experiments with cell extracts indicated that an alcohol dehydrogenase and an aldehyde dehydrogenase were also involved in the degradation of 2-CVE. This suggests that 2-CVE is metabolized via 2-hydroxyethylvinylether and vinyloxyacetaldehyde to vinyloxyacetic acid. Enzymatic ether cleavage was not detected. 2-CVE was also degraded by chemical ether cleavage, leading to the formation of 2-chloroethanol and acetaldehyde, both of which supported growth. We propose that A. aquaticus strains may be important for the detoxification and degradation of halogenated aliphatic compounds in the environment. PMID- 16349033 TI - Improved aerobic colony count technique for hydrophobic grid membrane filters. AB - The AOAC International official action procedure for performing aerobic colony counts on hydrophobic grid membrane filters (HGMFs) uses Trypticase soy-fast green FCF agar (FGA) incubated for 48 h. Microbial growths are various shades of green on a pale green background, which can cause problems for automated as well as manual counting. HGMFs which had been incubated 24 or 48 h at 35 degrees C on Trypticase soy agar were flooded underneath with 1 to 2 ml of 0.1% triphenyltetrazolium chloride (TTC) solution by simply lifting one corner of the filter while it was still on the agar and adding the reagent. Microbial growths on HGMFs were counted after color had been allowed to develop for 15 min at room temperature. With representative foods, virtually all colonies stained pink to red. Automated electronic counts made by using the MI-100 HGMF Interpreter were easier and more reliable than control HGMF counts made by the AOAC International official action procedure. Manual counting was easier as well because of increased visibility of the microbial growths. Except in the case of dairy products, 24-h TTC counts did not differ significantly from 48-h FGA counts, whereas the FGA counts at 24 h were always significantly lower, indicating that for many food products the HGMF TTC flooding method permits aerobic colony counts to be made after 24 h. PMID- 16349034 TI - Purification and Characterization of Maleate Hydratase from Pseudomonas pseudoalcaligenes. AB - Maleate hydratase (malease) from Pseudomonas pseudoalcaligenes has been purified. The purified enzyme (98% pure) catalyzes the stereospecific addition of water to maleate and citraconate (2-methylmaleate), forming d-(+)-malate and d-(+) citramalate, respectively. 2,3-Dimethylmaleate was also a substrate for malease. The stability of the enzyme was dependent on the protein concentration and the addition of dicarboxylic acids. The purified enzyme (89 kDa) consisted of two subunits (57 and 24 kDa). No cofactor was required for full activity of this colorless enzyme. Maximum enzyme activity was measured at pH 8 and 45 degrees C. The K(m) for maleate was 0.35 mM, and that for citraconate was 0.20 mM. Thiol reagents, such as p-chloromercuribenzoate and iodoacetamide, and sodium dodecyl sulfate completely inhibited malease activity. Malease activity was competitively inhibited by d-malate (K(i) = 0.63 mM) and d-citramalate (K(i) = 0.083 mM) and by the substrate analog 2,2-dimethylsuccinate (K(i) = 0.025 mM). The apparent equilibrium constants for the maleate, citraconate, and 2,3-dimethylmaleate hydration reactions were 2,050, 104, and 11.2, respectively. PMID- 16349035 TI - Identification and Cloning of Genes Involved in Specific Desulfurization of Dibenzothiophene by Rhodococcus sp. Strain IGTS8. AB - The gram-positive bacterium Rhodococcus sp. strain IGTS8 is able to remove sulfur from certain aromatic compounds without breaking carbon-carbon bonds. In particular, sulfur is removed from dibenzothiophene (DBT) to give the final product, 2-hydroxybiphenyl. A genomic library of IGTS8 was constructed in the cosmid vector pLAFR5, but no desulfurization phenotype was imparted to Escherichia coli. Therefore, IGTS8 was mutagenized, and a new strain (UV1) was selected that had lost the ability to desulfurize DBT. The genomic library was transferred into UV1, and several colonies that had regained the desulfurization phenotype were isolated, though free plasmid could not be isolated. Instead, vector DNA had integrated into either the chromosome or a large resident plasmid. DNA on either side of the inserted vector sequences was cloned and used to probe the original genomic library in E. coli. This procedure identified individual cosmid clones that, when electroporated into strain UV1, restored desulfurization. When the origin of replication from a Rhodococcus plasmid was inserted, the efficiency with which these clones transformed UV1 increased 20- to 50-fold and they could be retrieved as free plasmids. Restriction mapping and subcloning indicated that the desulfurization genes reside on a 4.0-kb DNA fragment. Finally, the phenotype was transferred to Rhodococcus fascians D188-5, a species normally incapable of desulfurizing DBT. The mutant strain, UV1, and R. fascians produced 2-hydroxybiphenyl from DBT when they contained appropriate clones, indicating that the genes for the entire pathway have been isolated. PMID- 16349036 TI - Construction of a Biotin-Overproducing Strain of Serratia marcescens. AB - We have isolated mutants resistant to acidomycin, a biotin analog, from Serratia marcescens Sr41. Strain SB304, resistant to 0.5 mg of acidomycin (frequently called actithiazic acid) per ml, produced 5 mg of d-biotin per liter of a medium containing sucrose and urea. Strain SB412, which was isolated from SB304 on a minimal agar plate containing 2 mg of acidomycin per ml and 0.1 mg of 5-(2 thienyl)-valeric acid per ml, produced 20 mg of d-biotin per ml. The two enzymes related to biotin synthesis were found to be released from biotin-mediated feedback repression in these mutants. Transductional analysis revealed that SB412 had acquired at least two mutations, one in the biotin operon locus and the other in an unknown locus distant from the biotin operon locus. PMID- 16349037 TI - Thermostability of Fumonisin B(1), a Mycotoxin from Fusarium moniliforme, in Corn. AB - Fumonisin B(1) (FB(1)) is a mycotoxin from Fusarium moniliforme that is frequently associated with corn. Thermal treatments are used in many processes concerning this cereal and its derivatives. The thermostability of this toxin in dry contaminated corn, resulting from F. moniliforme culture, was studied in different time-temperature combinations. FB(1) was quantified by instrumentalized thin-layer chromatography after a two-step sequential development and postchromatographic derivatization by p-anisaldehyde. The identity of FB(1) in extracts, before and after heat treatments, was confirmed by high-pressure liquid chromatography. For each temperature, the natural logarithm of the ratio of resulting FB(1) on initial content (In C/C(0)) is linearly correlated to exposure time. The calculated half-lives (L(50)), corresponding to the 50% value, were 10 min, 38 min, 175 min, and 8 h at 150, 125, 100, and 75 degrees C, respectively. There is a linear relationship between calculated L(50)s on a logarithmic scale and temperature. Therefore FB(1) is not significantly destroyed by the main drying processes of corn or thermal treatments used for its derivatives. Other associated means are required for detoxification. PMID- 16349038 TI - Application of a tetrazolium salt with a water-soluble formazan as an indicator of viability in respiring bacteria. AB - The tetrazolium salt sodium 3'-{1-[(phenylamino)-carbonyl]-3,4-tetrazolium}-bis (4-methoxy-6-nitro)benzene-sulfonic acid hydrate (XTT) was examined for use as a colorimetric indicator of viability in respiring bacteria. XTT was reduced to an orange, water-soluble formazan product by Methylosinus trichosporium OB3b, Pseudomonas putida, Escherichia coli, and Bacillus subtilis. Formazan production was proportional to live cell biomass, and XTT was reduced by all cultures in the absence of added electron-coupling agents. XTT reduction by M. trichosporium OB3b was linear over several hours and was stimulated by the presence of an exogenous substrate (methanol). Addition of cyanide to cultures incubated under oxic conditions gave an initial 10-fold increase in XTT reduction. Viability of bacteria incubated in the absence of exogenous carbon substrates was measured as XTT reduction and compared with viability estimates from plate counts. Results obtained with the two methods were generally comparable, but the XTT assay was superior when cell recovery on plates was low. Incubation of E. coli for 7 days in the absence of exogenous carbon substrates decreased viability by 90%, whereas the corresponding decreases for cultures of M. trichosporium OB3b, P. putida, and B. subtilis were less than 40%. PMID- 16349039 TI - Mineralization of 2,4-Dichlorophenoxyacetic Acid (2,4-D) and Mixtures of 2,4-D and 2,4,5-Trichlorophenoxyacetic Acid by Phanerochaete chrysosporium. AB - Evidence is presented for mineralization of 2,4-dichlorophenoxyacetic acid (2,4 D) in nutrient-rich media (high-nitrogen and malt extract media) by wild-type Phanerochaete chrysosporium and by a peroxidase-negative mutant of this organism. Mass balance analysis of [U-ring-C]2,4-D mineralization in malt extract cultures showed 82.7% recovery of radioactivity. Of this, 38.6% was released as CO(2) and 27.0, 11.2, and 5.9% were present in the aqueous, methylene chloride, and mycelial fractions, respectively. 2,4-D and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) were simultaneously mineralized when presented as a mixture, and mutual inhibition of degradation was not observed. In contrast, a relatively higher rate of mineralization of 2,4-D and 2,4,5-T was observed when these compounds were tested as mixtures than when they were tested alone. PMID- 16349040 TI - Metabolism of Linoleic Acid or Mevalonate and 6-Pentyl-alpha-Pyrone Biosynthesis by Trichoderma Species. AB - The understanding of the biosynthetic pathway of 6-pentyl-alpha-pyrone in Trichoderma species was achieved by using labelled linoleic acid or mevalonate as a tracer. Incubation of growing cultures of Trichoderma harzianum and T. viride with [U-C]linoleic acid or [5-C]sodium mevalonate revealed that both fungal strains were able to incorporate these labelled compounds (50 and 15%, respectively). Most intracellular radioactivity was found in the neutral lipid fraction. At the initial time of incubation, the radioactivity from [C]linoleic acid was incorporated into 6-pentyl-alpha-pyrone more rapidly than that from [C]mevalonate. No radioactivity incorporation was detected in 6-pentyl-alpha pyrone when fungal cultures were incubated with [1-C]linoleic acid. These results suggested that beta-oxidation of linoleic acid was a probable main step in the biosynthetic pathway of 6-pentyl-alpha-pyrone in Trichoderma species. PMID- 16349041 TI - Chemotaxis of azospirillum species to aromatic compounds. AB - Chemotaxis of Azospirillum lipoferum Sp 59b and Azospirillum brasilense Sp 7 and Sp CD to malate and to the aromatic substrates benzoate, protocatechuate, 4 hydroxybenzoate, and catechol was assayed by the capillary method and direct cell counts. A. lipoferum required induction by growth on 4-hydroxybenzoate for positive chemotaxis to this compound. Chemotaxis of Azospirillum spp. to all other substrates did not require induction. Maximum chemotactic responses for most aromatic compounds occurred at concentrations of 1 to 10 mM for A. lipoferum and 100 muM to 1 mM for A. brasilense. Threshold levels of these chemoattractants ranged from nanomolar to micromolar, with A. brasilense Sp CD showing the lowest threshold levels for the substrates tested. Benzoate was the strongest chemoattractant tested, with threshold concentrations in the nanomolar to picomolar range for all strains. Azospirillum spp. clearly have more sensitive chemosensory mechanisms for certain aromatic substrates than previously reported in some other soil bacteria. This sensitivity allows Azospirillum spp. to detect and respond to aromatic substrates at concentrations relevant to the soil and rhizosphere environments. The ability to detect such low concentrations of aromatic compounds in soils may confer advantages in survival and colonization of the rhizosphere by Azospirillum species. PMID- 16349042 TI - Phenotypic Characteristics and Virulence of Vibrio anguillarum-Related Organisms. AB - The phenotypic, molecular, and virulence properties of 46 Vibrio anguillarum related (VAR) strains isolated from diseased fish and shellfish and from the environment were investigated. Twelve reference strains belonging to the 10 serotypes of V. anguillarum and the Vibrio splendidus type strain were included for comparison. Numerical taxonomy studies allowed us to group the isolates into four phena. The main phenotypic traits to differentiate VAR strains from V. anguillarum were fermentation of arabinose and mannitol, indole and Voges Proskauer reactions, gelatin and casein hydrolysis, hemolytic activity, growth at 37 and 4 degrees C, and resistance to ampicillin. Serological analysis confirmed that phena I and II were composed mainly of strains of V. anguillarum, while phena III and IV included VAR strains. Excluding the reference strains, the typeable isolates belonged to serotypes O3 (15 strains), O4 (3 strains), and O5 (2 strains) of V. anguillarum. The infectivity trials showed that only 9 of a total of 24 strains tested displayed virulence for rainbow trout. Virulent strains (50% lethal dose ranging from 10 to 10 cells) included V. anguillarum strains belonging to serotypes O1 (one strain), O2 (one strain), O3 (three isolates), and O4 (one isolate) and only three strains of the VAR group. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of lipopolysaccharide and outer membrane proteins showed heterogeneity not only among the 10 V. anguillarum serotypes but also within the VAR group. Immunoblot assays demonstrated a close relationship among V. anguillarum strains from the same serotype, while strains from different serotypes were not antigenically related. The VAR strains did not share antigenic components with the serotypes of V. anguillarum tested (serotypes O1 to O5). Plasmids were detected in only 19 of the total of 59 strains. The majority of the strains carrying plasmids were grouped within phenon IV, in which plasmid bands of 27 and 36 MDa were found in all the isolates. No correlation between the plasmid content of VAR microorganisms and their phenotypic or virulence characteristics was observed. From these results it can be concluded that VAR strains associated with disease should be included together with V. anguillarum in the formulation of vaccines against vibriosis. PMID- 16349043 TI - Uncoupler-Resistant Glucose Uptake by the Thermophilic Glycolytic Anaerobe Thermoanaerobacter thermosulfuricus (Clostridium thermohydrosulfuricum). AB - The transport of glucose across the bacterial cell membrane of Thermoanaerobacter thermosulfuricus (Clostridium thermohydrosulfuricum) Rt8.B1 was governed by a permease which did not catalyze concomitant substrate transport and phosphorylation and thus was not a phosphoenolpyruvate-dependent phosphotransferase. Glucose uptake was carrier mediated, could not be driven by an artificial membrane potential (Deltapsi) in the presence or absence of sodium, and was not sensitive to inhibitors which dissipate the proton motive force (Deltap; tetrachlorosalicylanilide, N,N-dicyclohexylcarboiimide, and 2,4 dinitrophenol), and no uptake of the nonmetabolizable analog 2-deoxyglucose could be demonstrated. The glucokinase apparent K(m) for glucose (0.21 mM) was similar to the K(t) (affinity constant) for glucose uptake (0.15 mM), suggesting that glucokinase controls the rate of glucose uptake. Inhibitors of ATP synthesis (iodoacetate and sodium fluoride) also inhibited glucose uptake, and this effect was due to a reduction in the level of ATP available to glucokinase for glucose phosphorylation. These results indicated that T. thermosulfuricus Rt8.B1 lacks a concentrative uptake system for glucose and that uptake is via facilitated diffusion, followed by ATP-dependent phosphorylation by glucokinase. In T. thermosulfuricus Rt8.B1, glucose is metabolized by the Embden-Meyerhof-Parnas pathway, which yields 2 mol of ATP (G. M. Cook, unpublished data). Since only 1 mol of ATP is used to transport 1 mol of glucose, the energetics of this system are therefore similar to those found in bacteria which possess a phosphotransferase. PMID- 16349045 TI - Effects of polychlorinated biphenyl congener concentration and sediment supplementation on rates of methanogenesis and 2,3,6-trichlorobiphenyl dechlorination in an anaerobic enrichment. AB - We have employed a method of enrichment that allows us to significantly increase the rate of reductive polychlorinated biphenyl (PCB) dechlorination. This method shortens the time required to investigate the effects that culture conditions have on dechlorination and provides an estimate of the potential activity of the PCB-dechlorinating anaerobes. The periodic supplementation of sterile sediment and PCB produced an enhanced, measurable, and sustained rate of dechlorination. We observed volumetric rates of the dechlorination of 2,3,6-trichlorobiphenyl (2,3,6-CB) to 2,6-dichlorobiphenyl (2,6-CB) of more than 300 mumol liter day when the cultures were supplemented daily. A calculation of this activity that is based on an estimate of the number of dechlorinating anaerobes present indicates that 1.13 pmol of 2,3,6-CB was dechlorinated to 2,6-CB day bacterial cell. This rate is similar to that of the reductive dechlorination of 3-chlorobenzoate by Desulfomonile tiedjei. Methanogenesis declined from 585.3 to 125.9 mumol of CH(4) liter day, while dechlorination increased from 8.2 to 346.0 mumol of 2,3,6-CB dechlorinated to 2,6-CB liter day. PMID- 16349044 TI - Common mechanistic action of bacteriocins from lactic Acid bacteria. AB - The influence of four bacteriocins from lactic acid bacteria on the proton motive force (PMF) of sensitive organisms was investigated. Pediocin PA-1 (20 mug/ml) and leuconocin S (48.5 mug/ml) mediated total or major PMF dissipation of energized Listeria monocytogenes Scott A cells in a concentration-dependent manner, as has been shown for nisin. Lactacin F (13.5 mug/ml) caused 87% PMF depletion of energized Lactobacillus delbrueckii ATCC 4797 cells, also in a concentration-dependent fashion. The energy requirements for the activity of these four bacteriocins were determined by using the ionophores nigericin and valinomycin to carry out partial and specific deenergization of the target organisms. Pediocin PA-1, leuconocin S, and lactacin F acted in an energy independent manner, whereas the activity of nisin was confirmed to be energy dependent. These results together with published reports on other bacteriocins suggest that the bacteriocins of lactic acid bacteria share a common mechanism, the depletion of PMF. PMID- 16349046 TI - Localization of the Mating Type Gene in Agaricus bisporus. AB - The cultivated mushroom Agaricus bisporus is secondarily homothallic. Most basidia produce two basidiospores, each of which receives two of the four postmeiotic nuclei. Usually, the two packaged nuclei carry compatible mating types. Previous studies suggested that there may be only a single mating type locus in A. bisporus. In this study, we determined whether the mating type segregated as a single Mendelian determinant in a cross marked with 64 segregating molecular markers. To score mating types, each of the 52 homokaryotic offspring from this cross was paired with each of the two progenitor homokaryons. Compatible matings were identified by the formation of genetically stable heterokaryons which were verified by assay of restriction fragment length polymorphisms (RFLPs). Data for screening mycelial interactions on petri plates as well as fruit body formation were compared with the RFLP results. Mating types of 43 of the 52 homokaryotic offspring were determined on the basis of RFLP analysis. Our results indicate (i) there is a segregating mating type gene in A. bisporus, (ii) this mating type gene is on the largest linkage group (chromosome I), (iii) mycelial interactions on petri plates were associated with heterokaryon formation under selected conditions, (iv) fruit body formation was dependent upon the mating type gene, and (v) compatible mating types may not always be sufficient for fruiting. PMID- 16349047 TI - Bacteriophage Lambda as a Delivery Vector for Tn10-Derived Transposons in Xenorhabdus bovienii. AB - Xenorhabdus bovienii wild-type strains lack a functional receptor protein (LamB) in the outer membrane and as a result are unable to adsorb coliphage lambda (lambda). Introduction of plasmids encoding lamB into X. bovienii T228 results in constitutive expression of LamB in the outer membrane of this organism. LamB expressing strains of X. bovienii adsorb lambda bacteriophage particles and can be used as hosts for lambda::Tn constructs. A Tn10-derived transposon, element 9 (J. C. Way, D. Davis, D. Morisato, D. E. Roberts, and N. Kleckner, Gene 32:369 379, 1984) was used to construct a variety of insertion mutants of X. bovienii. Mutants that had altered expression of protease, lipase, DNase, dye-binding capability, and hemolytic activity, in addition to a series of auxotrophic mutants, were isolated. PMID- 16349048 TI - Oxalate- and Glyoxylate-Dependent Growth and Acetogenesis by Clostridium thermoaceticum. AB - The acetogenic bacterium Clostridium thermoaceticum ATCC 39073 grew at the expense of the two-carbon substrates oxalate and glyoxylate. Other two-carbon substrates (acetaldehyde, acetate, ethanol, ethylene glycol, glycolaldehyde, glycolate, and glyoxal) were not growth supportive. Growth increased linearly with increasing substrate concentrations up to 45 mM oxalate and glyoxylate, and supplemental CO(2) was not required for growth. Oxalate and glyoxylate yielded 4.9 and 9.4 g, respectively, of cell biomass (dry weight) per mol of substrate utilized. Acetate was the major reduced end product recovered from oxalate and glyoxylate cultures. C labeling studies showed that oxalate was subject to decarboxylation, and product analysis indicated that oxalate was utilized by the following reaction: 4OOC-COO + 5H(2)O --> CH(3)COO + 6HCO(3) + OH. Oxalate- and glyoxylate-dependent growth produced lower acetate concentrations per unit of cell biomass synthesized than did H(2)-, CO-, methanol-, formate-, O-methyl-, or glucose-dependent growth. Protein profiles of oxalate-grown cells were dissimilar from protein profiles of glyoxylate-, CO-, or formate-grown cells, suggesting induction of new proteins for the utilization of oxalate. C. thermoaceticum DSM 2955 and Clostridium thermoautotrophicum JW 701/3 also grew at the expense of oxalate and glyoxylate. However, oxalate and glyoxylate did not support the growth of C. thermoaceticum OMD (a nonautotrophic strain) or six other species of acetogenic bacteria tested. PMID- 16349049 TI - 4-Methylumbelliferyl-beta-N-Acetylglucosaminide Hydrolysis by a High-Affinity Enzyme, a Putative Marker of Protozoan Bacterivory. AB - Hydrolysis of an artificial fluorogenic substrate, 4-methylumbelliferyl-beta-N acetylglucosaminide, has been studied in a monoculture predator-prey system with either a flagellate (Bodo saltans) or a ciliate (Cyclidium sp.) fed upon pure bacterial culture (Aeromonas hydrophila or Alcaligenes xylosoxidans). Aeromonas hydrophila produced a low-affinity beta-N-acetylglucosaminidase-like enzyme (K(m), >>100 mumol liter) but Alcaligenes xylosoxidans did not. Inoculation of both bacterial strains with bacterivorous protozoa induced the occurrence of another, high-affinity, beta-N-acetylglucosaminidase-like enzyme (K(m), <0.5 mumol liter). The latter enzyme showed significant, close correlations with total grazing rates of both B. saltans (r = 0.96) and Cyclidium sp. (r = 0.89) estimated by using uptake of fluorescently labelled bacteria. Further significant correlations between several protozoan parameters and kinetic parameters of this enzyme suggest its likely protozoan origin. If both types of enzyme occurred together, they could be satisfactorily distinguished by using kinetic data analysis. Hence, measurements of beta-N-acetylglucosaminidase-like activities might be promising to use to improve estimations of protozoan bacterivory. PMID- 16349050 TI - Corrinoid-Dependent Methyl Transfer Reactions Are Involved in Methanol and 3,4 Dimethoxybenzoate Metabolism by Sporomusa ovata. AB - Washed and air-oxidized proteins from Sporomusa ovata cleaved the C-O bond of methanol or methoxyaromatics and transferred the methyl to dl-tetrahydrofolate. The reactions strictly required a reductive activation by titanium citrate, catalytic amounts of ATP, and the addition of dl-tetrahydrofolate. Methylcorrinoid-containing proteins carried the methanol methyl, which was transferred to dl-tetrahydrofolate at a specific rate of 120 nmol h mg of protein. Tetrahydrofolate methylation diminished after the addition of 1 iodopropane or when the methyl donor methanol was replaced by 3,4 dimethoxybenzoate. However, whole Sporomusa cells utilize the methoxyl groups of 3,4-dimethoxybenzoate as a carbon source by a sequential O demethylation to 4 hydroxy-3-methoxybenzoate and 3,4-dihydroxybenzoate. The in vitro O demethylation of 3,4-[4-methoxyl-C]dimethoxybenzoate proceeded via two distinct corrinoid containing proteins to form 5-[C]methyltetrahydrofolate at a specific rate of 200 nmol h mg of protein. Proteins from 3,4-dimethoxybenzoate-grown cells efficiently used methoxybenzoates with vicinal substituents only, but they were unable to activate methanol. These results emphasized that specific enzymes are involved in methanol activation as well as in the activation of various methoxybenzoates and that similar corrinoid-dependent methyl transfer pathways are employed in 5 methyl-tetrahydrofolate formation from these substrates. Methyl-tetrahydrofolate could be demethylated by a distinct methyl transferase. That enzyme activity was present in washed and air-oxidized cell extracts from methanol-grown cells and from 3,4-dimethoxybenzoate-grown cells. It used cob(I)alamin as the methyl acceptor in vitro, which was methylated at a rate of 48 nmol min mg of protein even when ATP was omitted from the assay mixture. This methyl-cob(III)alamin formation made possible a spectrophotometric quantification of the preceding methyl transfers from methanol or methoxybenzoates to dl-tetrahydrofolate. PMID- 16349051 TI - Direct quantification of unadsorbed viruses in suspensions of adsorbing colloids with the enzyme-linked immunosorbent assay. AB - Quantification of two plant viruses in suspensions of homoionic Ca-bentonite was conducted by applying to the enzyme-linked immunosorbent assay plates either the virus-clay mixture (direct method) or the supernatant obtained after the clay was allowed to settle (classic method). Both methods showed a similar dependence of free virus content on clay concentration. A higher content at equilibrium was measured for both viruses by the direct method. The advantage of using the direct over the classic method is discussed. PMID- 16349052 TI - Characterization by Multilocus Enzyme Electrophoresis of Listeria monocytogenes Isolates Involved in Ovine Listeriosis Outbreaks in Scotland from 1989 to 1991. AB - Initial results from a study of five small ovine listeriosis outbreaks in Scotland in 1989 to 1991 are presented. Forty-eight isolates including three from silage were typed at 10 polymorphic enzyme loci by using multilocus enzyme electrophoresis resulting in the identification of 12 electrophoretic types. Phylogenetic analysis partitioned the 12 electrophoretic types into two statistically distinct divisions distinguishing 1/2a serotypes from non-1/2a serotypes. PMID- 16349053 TI - Genotypic Diversity among Strains of Bradyrhizobium japonicum Belonging to Serogroup 110. AB - Thirty-three strains of Bradyrhizobium japonicum within serogroup 110 were examined for genotypic diversity by using DNA-DNA hybridization analyses. The analysis of the DNA from 15 hydrogen-uptake-negative strains with the bradyrhizobial uptake hydrogenase probe pHU52 showed variation in degree of homology and restriction fragment length polymorphism of EcoRI-restricted DNA. Clustering analysis of the 33 strains on the basis of DNA-DNA hybridization analysis with four restriction enzymes and with the bradyrhizobial nodulation locus, pRJUT10, as probe indicated the existence of four groups of strains, which were less than 70% similar. Restriction digestion of genomic DNA with BamHI and DNA-DNA hybridization with pRJUT10 permitted classification of each of the strains according to a specific fingerprint pattern. PMID- 16349054 TI - Purification and Properties of a Highly Active Organophosphorus Acid Anhydrolase from Alteromonas undina. AB - A highly active organophosphorus acid anhydrolase from Alteromonas undina was purified to homogeneity and found to be composed of a single polypeptide chain with a molecular weight of 53,000. With diisopropylfluorophosphate as a substrate, the purified enzyme has a specific activity of approximately 575 mumol/min/mg of protein. The enzyme has optimum activity at pH 8.0 and 55 degrees C and is stimulated by sulfhydryl reducing agents and manganese. It is capable of rapidly hydrolyzing a wide range of nerve agents and several chromogenic phosphinates. PMID- 16349055 TI - Colonization of Crystalline Cellulose by Clostridium cellulolyticum ATCC 35319. AB - Cellulose colonization by Clostridium cellulolyticum was studied by using [methyl H]thymidine incorporation. The colonization process indicated that a part of the bacterial population was released from cellulose to the liquid phase before binding and colonizing another adhesion site of the cellulose. We postulate that cellulose colonization occurs according to the following process: adhesion, colonization, release, and readhesion. PMID- 16349056 TI - Effect of Glucose Utilization on Nitrite Excretion by the Unicellular Cyanobacterium Synechocystis sp. Strain PCC 6803. AB - Up to 1 mM nitrite was excreted by Synechocystis strain 6803 cells growing under mixotrophic or photoheterotrophic conditions. This excretion is not due to a lower ratio of nitrite and nitrate reductase activities in the presence of glucose but seems to be related to a shortage of reduced ferredoxin, their electron donor, as a result of a decrease in noncyclic photosynthetic flow observed under these circumstances. Because about 60% of the reduced nitrate is excreted, the potential utilization of cyanobacteria for removal of nitrate from contaminated waters containing high concentrations of organic compounds is questioned. PMID- 16349057 TI - In Vitro Stimulation of Forage Fiber Degradation by Ruminal Microorganisms with Aspergillus oryzae Fermentation Extract. AB - Aspergillus oryzae fermentation extract (Amaferm) was evaluated for its ability to influence degradation of brome grass and switchgrass fiber fractions by mixed ruminal microorganisms in vitro. Addition of Amaferm at a concentration of 0.067 mg/ml, which is approximately the concentration found in the rumen ecosystem (0.06 mg/ml), increased the degradation of brome grass neutral detergent fiber (NDF) by 28% after fermentation for 12 h (P < 0.01), but had no effect after fermentation for 24 or 48 h. The levels of degradation of both the cellulose and hemicellulose fractions were increased after fermentation for 12 h (P < 0.01). Additions of 0.08 and 8% (vol/vol) Amaferm filtrate (12.5 g/100 ml) stimulated degradation of switchgrass NDF by 12 and 24% (P < 0.01), respectively, after fermentation for 12 h; when 80% filtrate was added, degradation was decreased by 38%. The concentrations of total anaerobes in culture tubes containing 80% filtrate were 5 times greater than the concentrations in the controls; however, the concentrations of cellulolytic organisms were 3.5 times lower than the concentrations in the controls (P < 0.05). These results suggested that the filtrate contained high concentrations of soluble substrate which did not allow the cellulolytic organisms to compete well with other populations. The remaining concentrations of esterified p-coumaric and ferulic acids were lower at 12 h in NDF residues obtained from fermentation mixtures supplemented with Amaferm. Because the total anaerobes were not inhibited in fermentation mixtures containing Amaferm, antibiotics are unlikely to be involved as a mode of action for increasing NDF degradation. The possibility that Amaferm contains enzymes (possibly esterases) that may play a role in stimulating the rate of fiber degradation by mixed ruminal microorganisms by removal of plant cell wall phenolic acid esters is discussed. PMID- 16349058 TI - Characterization of Loosely Associated Material from the Cell Surface of Lactococcus lactis subsp. cremoris E8 and Its Phage-Resistant Variant Strain 398. AB - Loosely associated material (LAM) was isolated by gentle extraction procedures from the cell surface of Lactococcus lactis subsp. cremoris E8 and its phage resistant variant strain 398. LAM from both strains was chemically characterized, and its role in the adsorption of three small isometric bacteriophages, phi 618, phi 833, and phi 852, to the cell surface of the two strains was investigated. The phage-resistant strain (strain 398) produced LAM which differed significantly from the material produced by the parent strain. The total yield of LAM from strain 398 was two- to threefold higher than that from strain E8, and the material contained fivefold more rhamnose and twofold more galactose. Polyacrylamide gel electrophoretic analysis showed that LAM from strain 398 lacked a 21-kDa protein which was present in LAM from the parent strain. Inhibition studies of phage binding by using isolated LAM from two strains showed that although LAM from strain E8 reduced the titer of phi 618 and phi 852 by 53 and 82% respectively, LAM from strain 398 had no effect on the plaque-forming ability of any of the three phages tested. Treatment of LAM from strain E8 with sodium metaperiodate destroyed its ability to bind with phi 618 and phi 852. Phenotypically, strain 398 differed from its parent strain E8 in that it was more prone to cell lysis and required an osmotically adjusted buffer system for the extraction of LAM. PMID- 16349059 TI - Dormancy in Stationary-Phase Cultures of Micrococcus luteus: Flow Cytometric Analysis of Starvation and Resuscitation. AB - Cultures of the copiotrophic bacterium Micrococcus luteus were stored in spent growth medium for an extended period of time following batch culture. After an initial decrease, the total cell counts remained constant at approximately 60 to 70% of the counts at the beginning of storage. The level of viability, as judged by plate counts, decreased to less than 0.05%, while respiration and the ability to accumulate the lipophilic cation rhodamine 123 decreased to undetectable levels. However, using penicillin pretreatment (to remove viable cells) and flow cytometry and by monitoring both the total and viable counts, we found that at least 50% of the cells in populations of 75-day-old cultures were not dead but were dormant. Resuscitation in liquid medium was accompanied by the appearance of a population of larger cells, which could accumulate rhodamine 123 and reduce the dye 5-cyano-2,3-ditolyl tetrazolium chloride to a fluorescent formazan, while a similar fraction of the population was converted to colony-forming, viable cells. We surmise that dormancy may be far more common than death in starving microbial cultures. PMID- 16349060 TI - Nuclear Ribosomal DNA as a Probe for Genetic Variability in the Japanese Pear Pathotype of Alternaria alternata. AB - A restriction fragment length polymorphism analysis of nuclear ribosomal RNA genes (rDNA) was used to measure the amount and distribution of genetic variability in populations of the Japanese pear pathotype of Alternaria alternata on both micro- and macrogeographical scales. A total of 322 isolates were obtained from 13 areas in Aichi, Gifu, and Tottori Prefectures in central and western Japan. The restriction fragment length polymorphism analysis revealed that the pathogen populations contained at least eight rDNA variants. The eight variant types differed in the lengths and in the presence of the restriction sites in spacer DNA outside the coding regions for rRNAs. A total of 271 isolates were classified into the eight types. The remaining 51 isolates were determined to have mixed rDNA types. Single pear fields typically contained two to five types of rDNA variants. The frequencies of rDNA variants in 11 populations in Tottori Prefecture were compared; in this prefecture orchards containing the susceptible pear are common. Except for one collection site, there were no significant differences in the composition of the rDNA variants among the populations. This suggests that dispersal of inocula has occurred frequently in Tottori Prefecture. In contrast, significantly different distributions were observed in the three prefectures, indicating that gene flow between prefectures might be limited by geographical isolation. DNA fingerprints resulting from hybridization with a moderately repetitive DNA sequence of the fungus revealed greater genetic variability and geographical differences in genetic population structure even within the same rDNA type. PMID- 16349061 TI - Two Uptake Systems for Fructose in Lactococcus lactis subsp. cremoris FD1 Produce Glycolytic and Gluconeogenic Fructose Phosphates and Induce Oscillations in Growth and Lactic Acid Formation. AB - Fructose transport in lactococci is mediated by two phosphotransferase systems (PTS). The constitutive mannose PTS has a broad specificity and may be used for uptake of fructose with a fructose saturation constant (K(Fru)) of 0.89 mM, giving intracellular fructose 6-phosphate. The inducible fructose PTS has a very small saturation constant (K(Fru), <17 muM), and the fructose 1-phosphate produced enters the Embden-Meyerhof-Parnas (EMP) pathway as fructose 1,6 diphosphate. Growth in batch cultures of Lactococcus lactis subsp. cremoris FD1 in a yeast extract medium with fructose as the only sugar is poor both with respect to specific growth rate and biomass yield, whereas the specific lactic acid production rate is higher than those in similar fermentations on other sugars metabolized via the EMP pathway, e.g., glucose. In fructose-limited chemostat cultures, the biomass concentration exhibits a strong correlation with the dilution rate, and starting a continuous culture at the end of a batch fermentation leads to large and persistent oscillations in the biomass concentration and specific lactic acid production rate. Two proposed mechanisms underlying this strange growth pattern follow. (i) Fructose transported via the fructose PTS cannot be converted into essential biomass precursors (glucose 6 phosphate or fructose 6-phosphate), because L. lactis subsp. cremoris FD1 is devoid of fructose 1,6-diphosphatase activity. (ii) The fructose PTS apparently produces a metabolite (presumably fructose 1-phosphate) which exerts catabolite repression of both mannose PTS and lactose PTS. Since the repressed mannose PTS and lactose PTS are shown to have identical maximum molar transport rates, the results indicate that it is the general PTS proteins which are repressed. PMID- 16349062 TI - Cytochrome Oxidase: Subcellular Distribution and Relationship to Nitrogenase Expression in the Nonheterocystous Marine Cyanobacterium Trichodesmium thiebautii. AB - Immunochemical labeling was used to study the subcellular distribution of cytochrome oxidase, a respiratory protein, in Trichodesmium thiebautii. The protein was found associated with both cytoplasmic and thylakoid membranes. About a sixfold variation in the protein content (gold particle count) was found among Trichodesmium cells within a single colony. Double labeling was performed with cytochrome oxidase and nitrogenase antisera. Regression analysis of gold particle counts per unit of cell area of cytochrome oxidase and nitrogenase showed a positive correlation (r = 0.911); cells with higher nitrogenase levels also had higher levels of cytochrome oxidase. The parallel expression of two proteins suggests that respiratory oxygen uptake may be involved in nitrogenase protection (respiratory protection) in Trichodesmium spp. PMID- 16349063 TI - Bacterial Adhesion under Static and Dynamic Conditions. AB - The deposition of various pseudomonads and coryneform bacteria with different hydrophobicities (water contact angles) and negative cell surface charges on negatively charged Teflon and glass surfaces was investigated. The levels of deposition varied between 5.0 x 10 and 1.6 x 10 cells cm and between 5.0 x 10 and 3.6 x 10 cells cm for dynamic column and static batch systems, respectively, indicating that there was a wide variation in physicochemical interactions. Batch and column results were compared in order to better distinguish between hydrodynamic and other system-dependent influences and method-independent physicochemical interactions. Despite the shorter suspension-solid contact time in columns (1 h) than in batch systems (4 h), the level of deposition (expressed as the number of cells that adhered) divided by the applied ambient cell concentration was 4.12 +/- 1.63 times higher in columns than in batch sytems for 15 of 22 strain-surface combinations studied. This demonstrates that transport of microbial particles from bulk liquid to surfaces is more efficient in dynamic columns (transport dominated by convection and diffusion) than in static batch systems (transport by diffusion only). The relative constancy of this ratio for the 15 combinations shows that physicochemical interactions affect adhesion similarly in the two systems. The deviating deposition behavior of the other seven strain-surface combinations could be attributed to method-dependent effects resulting from specific cell characteristics (e.g., to the presence of capsular polymers, to an ability to aggregate, to large cell sizes, or to a tendency to desorb after passage through an air-liquid interface). PMID- 16349064 TI - Attachment stimulates exopolysaccharide synthesis by a bacterium. AB - This study examined the hypothesis that solid surfaces may stimulate attached bacteria to produce exopolymers. Addition of sand to shake-flask cultures seemed to induce exopolymer synthesis by a number of subsurface isolates, as revealed by optical microscopy. Several additional lines of evidence indicated that exopolymer production by attached cells (in continuous-flow sand-packed columns) was greater than by their free-living counterparts. Total carbohydrates and extracellular polysaccharides, both normalized to cell protein, were greater (2.5 and 5-fold, respectively) for attached cells than for free-living cells. Also, adsorption of a polyanion-binding dye to the exopolymer fraction was sixfold greater for attached cells than for unattached cells. When surface-grown cells were resuspended in fresh medium, exopolymer production decreased to the level characteristic of unattached cells, which ruled out the possibility that attached cells comprised a subpopulation of sticky mucoid variants. The mechanism by which attachment stimulated exopolymer synthesis did not involve changes of the specific growth rate, growth stage, or limiting nutrient. PMID- 16349065 TI - Microscale distribution of nitrification activity in sediment determined with a shielded microsensor for nitrate. AB - Microprofiles of O(2) and NO(3) were measured simultaneously in freshwater sediment with microsensors which were completely free from electrical interference because of coaxial designs. Depth profiles of nitrification (NO(3) production) and denitrification (NO(3) consumption) were subsequently determined by computer simulation of the measured microprofiles. The nitrifying bacterial community responded very quickly to changes in environmental conditions, and new steady-state microprofiles of O(2) and NO(3) were usually approached within a few hours after perturbation. Nitrification started quickly after introduction of O(2) in previously anoxic layers, suggesting prolonged survival of the nitrifiers during anaerobiosis. Changes in the availability of O(2) and NH(4) greatly affected the nitrification profile, and there was a high rate of coupled nitrification-denitrification under conditions in which nitrification occurred right above the oxic-anoxic interface. Addition of C(2)H(2) rapidly removed the NO(3) peaks, indicating that NO(3) production was due mainly to autotrophic nitrification. PMID- 16349066 TI - Survival of denitrifiers in nitrate-free, anaerobic environments. AB - Experiments were undertaken to explain the occurrence of a high denitrification capacity in anaerobic, NO(3)-free habitats. Deep layers of freshwater sediments that were buried more than 40 years ago and digested sludge were the habitats studied. The denitrifier populations were 3.1 x 10 and 3.1 x 10 cells cm in deep sediments from a river and lake, respectively, and 5.3 x 10 cells cm in digested sludge. The denitrification capacities of the samples reflected the population densities. Strict anaerobic procedures were used to obtain the predominant isolates that would grow on anaerobic medium with NO(3). All strict anaerobes isolated failed to denitrify. All isolates that denitrified were aerobic, gram negative bacteria, particularly species of Pseudomonas and Alcaligenes. No detectable growth was observed when these strains were incubated with electron acceptors other than NO(3) or O(2). When representative isolates were added to sterile, O(2)- and NO(3)-free porewater from their original locations at their natural densities (10 cells cm), no change in viable population was noted over 3 months of incubation. Metabolic activity was demonstrated in these cells by slow formation of formazan granules when exposed to tetrazolium and by observation of motile cells. When [C]glucose was added to cell suspensions of the pseudomonads that had been starved for 3 months without electron acceptors (O(2) or NO(3)), C labeled products, including cell biomass, CO(2), and fermentation products, were produced. The high denitrification capacity of these anaerobic environments appears to be due to conventional respiratory denitrifiers. These organisms have the capacity for long-term survival without O(2) or NO(3) and appear to be capable of providing for their maintenance by carrying on a low level of fermentation. PMID- 16349067 TI - Isolation and characterization of a pseudomonas strain producing glutaryl-7 aminocephalosporanic Acid acylase. AB - Several screening methods were developed for the selection of Pseudomonas strains capable of hydrolyzing glutaryl-7-aminocephalosporanic acid to 7 aminocephalosporanic acid. An isolate exhibiting high acylase activity, designated BL072, was identified as a strain of Pseudomonas diminuta. It grew optimally at pH 7 to 8 and at a temperature of 32 to 40 degrees C, but acylase activity was highest when the strain was grown at 28 degrees C. Mutants of BL072 were generated by nitrosoguanidine treatment and screened for increased production of glutaryl-7-aminocephalosporanic acid acylase. A superior mutant gave a fourfold increase in acylase titer. The cell-associated acylase had similar activities against various glutaryl-cephems but had undetectable activity against cephalosporin C. This acylase may prove useful for the conversion of cephalosporin C to 7-aminocephalosporanic acid. PMID- 16349068 TI - Fusarium poae and Fusarium crookwellense, Fungi Responsible for the Natural Occurrence of Nivalenol in Hokkaido. AB - To determine the reasons for the natural occurrence of nivalenol in the northernmost area of Japan, scabby wheat was harvested from 19 crop fields in Hokkaido. Mycological surveys and analysis for mycotoxin contamination were performed. Among 13 wheat grain samples harvested in seven locations, 9, 2, and 6 samples were positive for deoxynivalenol, nivalenol, and zearalenone, respectively, at levels ranging from 0.03 to 1.28 mug/g, 0.04 to 1.22 mug/g, and 2 to 25 ng/g, respectively. The predominant Fusarium species of the scabby wheat examined were F. sporotrichioides, F. avenaceum, F. poae, and F. crookwellense. Fifteen of 48 F. poae isolates and all four F. crookwellense isolates were screened for the production of seven derivatives of trichothecenes and zearalenone respectively, on rice culture. One isolate of F. poae produced diacetoxyscirpenol alone (4.3 mug/g); seven produced nivalenol (1.3 to 23.8 mug/g), 4-acetylnivalenol (0.1 to 4.6 mug/g), and diacetoxyscirpenol (0.9 to 99.5 mug/g); and five produced nivalenol alone (0.4 to 3.5 mug/g). The remaining two isolates produced no trichothecenes. Zearalenone production was not found in any isolate of F. poae tested. All isolates of F. crookwellense produced nivalenol (0.9 to 22.5 mug/g), 4-acetylnivalenol (0.5 to 25.0 mug/g), and zearalenone (1.4 to 162.5 mug/g). From these results, it is apparent that deoxynivalenol and zearalenone, and occasionally nivalenol, occur naturally throughout Hokkaido, and it is suggested that nivalenol-producing F. poae and F. crookwellense strains are responsible for the natural contamination with nivalenol found in the northernmost area of Japan. Furthermore, it was found for the first time that several isolates of F. poae distributed in Hokkaido possessed the ability to produce both type A and type B trichothecenes. PMID- 16349069 TI - Comparison of ATP and ergosterol as indicators of fungal biomass associated with decomposing leaves in streams. AB - ATP and ergosterol were compared as indicators of fungal biomass associated with leaves decomposing in laboratory microcosms and streams. In all studies, the sporulation rates of the fungi colonizing leaves were also determined to compare patterns of fungal reproductive activity with patterns of mycelial growth. During leaf degradation, ATP concentrations exhibited significant, positive correlations with ergosterol concentrations in the laboratory and when leaves had been air dried prior to being submerged in a stream. However, when freshly shed leaves were submerged in a stream, concentrations of ATP and ergosterol were negatively correlated during degradation. This appeared to be due to the persistence of leaf derived ATP in freshly shed leaves during the first 1 to 2 weeks in the stream. Estimates of fungal biomass from ergosterol concentrations of leaf litter were one to three times those calculated from ATP concentrations. ATP, ergosterol, and sporulation data generally provided similar information about the fungi associated with decomposing leaves in streams during periods when fungi were growing. Ergosterol concentrations provide a more accurate indication of fungal biomass in situations in which other organisms make significant contributions to ATP pools. PMID- 16349070 TI - Direct electron microscopy study on the morphological diversity of bacteriophage populations in lake plusssee. AB - Direct electron microscopy of bacteriophages adsorbed to a carbon film without prior enrichment by specific host strains or concentration by physical or chemical methods was used to study the morphological diversity of natural bacteriophage assemblages in a North German lake. All samples contained a mixture of morphologically different tailed viruses, which were regarded as bacteriophages. Most of them had isometric heads and long noncontractile tails, belonging to morphotype B1 (Siphoviridae). In addition, members of morphotypes A1 (Myoviridae), B2 (Siphoviridae with elongated heads), and C1 (Podoviridae) were present in lower numbers. Only one cubic virus was detected, while no filamentous or pleomorphic phages were found. Up to 11 different phages per sample, and a total of 39 phages when all samples were considered together, could be distinguished by morphological criteria. The total number of phages was estimated to be on the order of 10/ml. PMID- 16349071 TI - Production and Excretion of Nod Metabolites by Rhizobium leguminosarum bv. trifolii Are Disrupted by the Same Environmental Factors That Reduce Nodulation in the Field. AB - Lipooligosaccharides (Nod metabolites) have been shown to be essential for the successful nodulation of legumes. In strains of Rhizobium leguminosarum bv. trifolii, Nod metabolites were detected predominantly within the cell and to a lesser extent in the periplasmic space and the growth medium. The production, and in particular the excretion, of Nod metabolites was restricted by a range of environmental conditions which are associated with poor nodulation in the field. Lowering the medium pH from 7.0 to 5.0, reducing the phosphate concentration from 1 mM to 5 muM KH(2)PO(4), and lowering the incubation temperature from 28 to 18 degrees C affected the number and relative concentrations of the Nod metabolites made. The form and concentration of the nitrogen source affected the relative concentrations of the Nod metabolites produced and excreted. KNO(3) concentrations of >10 mM did not affect cell growth rate but substantially reduced the number of Nod metabolites released. Environmental stresses differentially altered Nod metabolite production and excretion in the same strain carrying different introduced nod regions. Strain ANU845(pWLH1) produced and excreted comparatively fewer Nod metabolites at pH 5.0 and at 18 degrees C than strain ANU845(pRI4003). The excretion but not the production of Nod metabolites by strain ANU845(pRtO32) was dependent on the presence of both nodI and nodJ. Tn5 induced nodI and nodJ mutants did not accumulate any metabolites either outside the cell or within the outer membrane or periplasmic space. Recognition that Nod metabolite accumulation is a complex system of production and excretion, with each component responding differently to changes in environmental conditions, has many consequences, both at the molecular level and in the field. The ability of different strains to produce and release Nod metabolites is likely to be a major determinant of nodule occupancy and should be considered when screening strains suitable for adverse environments. PMID- 16349072 TI - Resistance to co-occurring phages enables marine synechococcus communities to coexist with cyanophages abundant in seawater. AB - Recent reports documenting very high viral abundances in seawater have led to increased interest in the role of viruses in aquatic environments and a resurgence of the hypothesis that viruses are significant agents of bacterial mortality. Synechococcus spp., small unicellular cyanobacteria that are important primary producers at the base of the marine food web, were used to assess this hypothesis. We isolated a diverse group of Synechococcus phages that at times reach titers of between 10 and 10 cyanophages per ml in both inshore and offshore waters. However, despite their diversity and abundance, we present evidence in support of the hypothesis that lytic phages have a negligible effect in regulating the densities of marine Synechococcus populations. Our results indicate that these bacterial communities are dominated by cells resistant to their co-occurring phages and that these viruses are maintained by scavenging on the relatively rare sensitive cells in these communities. PMID- 16349073 TI - Near-bottom pelagic bacteria at a deep-water sewage sludge disposal site. AB - The epibenthic bacterial community at deep-ocean sewage sludge disposal site DWD 106, located approximately 106 miles (ca. 196 km) off the coast of New Jersey, was assessed for changes associated with the introduction of large amounts of sewage sludge. Mixed cultures and bacterial isolates obtained from water overlying sediment core samples collected at the deep-water (2,500 m) municipal sewage disposal site were tested for the ability to grow under in situ conditions of temperature and pressure. The responses of cultures collected at a DWD-106 station heavily impacted by sewage sludge were compared with those of samples collected from a station at the same depth which was not contaminated by sewage sludge. Significant differences were observed in the ability of mixed bacterial cultures and isolates from the two sites to grow under deep-sea pressure and temperature conditions. The levels of sludge contamination were established by enumerating Clostridium perfringens, a sewage indicator bacterium, in sediment samples from the two sites. The results of hybridization experiments in which DNAs extracted directly from the water overlying sediment core samples were used indicate that the reference site epibenthic community, the disposal site epibenthic community, and the community in a surface sludge plume share many members. Decreased culturability of reference site mixed cultures in the presence of sewage sludge was observed. Thus, the culturable portions of both the autochthonous and allochthonous bacterial communities at the disposal site may be inhibited in situ, the former by sewage sludge and the latter by high pressure and low temperature. PMID- 16349074 TI - Invasion and Exclusion among Coexisting Pseudomonas syringae Strains on Leaves. AB - The invasion and exclusion abilities of coexisting Pseudomonas syringae strains were quantified on leaves. Twenty-nine P. syringae strains were inoculated onto plants in 107 pairwise combinations. All pairs were duplicated so that each strain was inoculated both first as an antagonist strain (day 0) and second as a challenge strain (day 3). The population size of each strain in a mixture was quantified on day 6 following incubation under moist conditions. For P. syringae strains, the presence of an established population often significantly reduced the growth of subsequently arriving challenge strains on the leaf surface. Invasion and exclusion abilities, quantified by contrasting population sizes of challenge strains in the presence and in the absence of another strain, varied significantly among P. syringae strains and were partly a function of the particular strain pair. The population size of a strain when present alone on a leaf was not predictive of invasion or exclusion ability. Successful invaders were significantly less likely to exclude challenge populations than were nonsuccessful invaders. Population sizes of successful excluders were negatively correlated with population sizes of coexisting challenge strains, while population sizes of successful invaders were positively correlated with those of coexisting antagonist strains. The patterns of interaction among coexisting strains suggest mechanisms for successful invasion and exclusion among P. syringae strains on leaves. PMID- 16349075 TI - Effects of Motility and Adsorption Rate Coefficient on Transport of Bacteria through Saturated Porous Media. AB - Three strains of Pseudomonas fluorescens with different motility rates and adsorption rate coefficients were injected into porous-medium reactors packed with l-mm-diameter glass spheres. Cell breakthrough, time to peak concentration, tailing, and cell recovery were measured at three interstitial pore velocities (higher than, lower than, and much lower than the maximal bacterial motility rate). All experiments were done with distilled water to reduce the effects of growth and chemotaxis. Contrary to expectations, motility did not result in either early breakthrough or early time to peak concentration at flow velocities below the motility rate. Bacterial size exclusion effects were shown to affect breakthrough curve shape at the very low flow velocity, but no such effect was seen at the higher flow velocity. The tendency of bacteria to adsorb to porous medium surfaces, as measured by adsorption rate coefficients, profoundly influenced transport characteristics. Cell recoveries were shown to be correlated with the ratio of advective to adsorptive transport in the reactors. Adsorption rate coefficients were found to be better predictors of microbial transport phenomena than individual characteristics, such as size, motility, or porous medium hydrodynamics. PMID- 16349076 TI - Induction and Regeneration of Autoplasts from Clostridium thermohydrosulfuricum JW102 and Thermoanaerobacter ethanolicus JW200. AB - Autolysis was induced to form stable, cell wall-free cells of Clostridium thermohydrosulfuricum JW102 and Thermoanaerobacter ethanolicus JW200, using a complex medium containing glycine (0.4% wt/vol) and/or sucrose or glycerol (10% wt/vol) at an optimum temperature of 64 degrees C. Autoplasts of both bacteria were grown as L-phase colonies on solid medium; more than 50% of these colonies regenerated to the walled form during prolonged incubation. The removal of the cell wall was confirmed by electron microscopy. PMID- 16349077 TI - Chromate Reduction by Resting Cells of Agrobacterium radiobacter EPS-916. AB - Resting cells of Agrobacterium radiobacter EPS-916 grown on glucose, fructose, maltose, lactose, mannitol, or glycerol reduced 0.5 mM chromate. However, resting cells of strain EPS-916 grown on glutamate or succinate did not reduce chromate. The ability of washed cells to reduce chromate was correlated with their redox potential. PMID- 16349078 TI - Microbial decomposition of elm and oak leaves in a karst aquifer. AB - Dry Chinquapin oak (Quercus macrocarpa) and American elm (Ulmus americana) leaves were placed in four microcosms fed by groundwater springs to monitor changes in dry mass, ash-free dry mass, and microbial activity over a 35-day period. Oxygen microelectrodes were used to measure microbial activity and to estimate millimeter-scale heterogeneity in that activity. Oak leaves lost mass more slowly than elm leaves. Generally, there was a decrease in total dry weight over the first 14 days, after which total dry weight began to increase. However, there were consistent decreases in ash-free dry mass over the entire incubation period, suggesting that the material remaining after initial leaf decomposition trapped inorganic particles. Microbial activity was higher on elm leaves than on oak leaves, with peak activity occurring at 6 and 27 days, respectively. The level of oxygen saturation on the bottom surface of an elm leaf ranged between 0 and 75% within a 30-mm area. This spatial heterogeneity in O(2) saturation disappeared when the water velocity increased from 0 to 6 cm s. Our results suggest that as leaves enter the groundwater, they decompose and provide substrate for microorganisms. The rate of decomposition depends on leaf type, small-scale variations in microbial activity, water velocity, and the length of submersion time. During the initial stages of decomposition, anoxic microzones are formed that could potentially be important to the biogeochemistry of the otherwise oxic aquifer. PMID- 16349079 TI - Variation in Nitrogen Utilization in Acremonium coenophialum Isolates. AB - Acremonium coenophialum, a fungal endosymbiont in tall fescue, is responsible for the production of alkaloid toxins. Animals grazing endophyte-infected tall fescue often show toxicosis. In marginal environments, the endophyte is important for long-term survival of tall fescue. Few differences in endophyte isolates from individual tall fescue plants have been reported. To aid development of a toxicosis-free tall fescue, it is important to identify differences in endophyte isolates. This report describes variation in nitrogen utilization in a defined culture medium by endophyte isolates from Kentucky-31 tall fescue. Overall, the best nitrogen sources for dry weight production of mycelium were proline and potassium nitrate. Thirty-four isolates grown on agar-solidified defined media with single nitrogen sources showed variation in nitrogen utilization. Fifty percent of the isolates were unable to utilize two or more amino acids. Manipulation of endophyte variation could lead to development of a toxicosis-free tall fescue cultivar. PMID- 16349080 TI - Phospholipid Fatty Acid composition, biomass, and activity of microbial communities from two soil types experimentally exposed to different heavy metals. AB - The phospholipid fatty acid (PLFA) pattern was analyzed in a forest humus and in an arable soil experimentally polluted with Cd, Cu, Ni, Pb, or Zn at different concentrations. In both soil types, there were gradual changes in the PLFA patterns for the different levels of metal contamination. The changes in the forest soil were similar irrespective of which metal was used, while in the arable soil the changes due to Cu contamination differed from those due to the other metals. Several PLFAs reacted similarly to the metal amendments in the two soil types, while others showed different responses. In both soils, the metal pollution resulted in a decrease in the iso-branched PLFAs i15:0 and i17:0 and in the monounsaturated 16:1omega5 and 16:1omega7c fatty acids, while increases were found for i16:0, the branched br17:0 and br18:0, and the cyclopropane cy17:0 fatty acids. In the forest soil, the methyl branched PLFAs 10Me16:0, 10Me17:0, and 10Me18:0 increased in metal-polluted soils, indicating an increase in actinomycetes, while in the arable soil a decrease was found for 10Me16:0 and 10Me18:0 in response to most metals. The bacterial PLFAs 15:0 and 17:0 increased in all metal-contaminated samples in the arable soil, while they were unaffected in the forest soil. Fatty acid 18:2omega6, which is considered to be predominantly of fungal origin, increased in the arable soil, except in the Cu amended samples, in which it decreased instead. Effects on the PLFA patterns were found at levels of metal contamination similar to or lower than those at which effects on ATP content, soil respiration, or total amount of PLFAs had occurred. PMID- 16349081 TI - Extra- and Intracellular Laccases of the Chestnut Blight Fungus, Cryphonectria parasitica. AB - A double-stranded RNA virus of the chestnut blight pathogen, Cryphonectria parasitica, has been shown previously to reduce accumulation of mRNAs of extracellular laccase (laccase A) produced by this fungus. Both extra- and intracellular laccases have been detected after growth of the fungus in liquid culture. In addition to cellular localization, the two laccases are distinguishable by time of appearance during growth and electrophoretic mobility. Laccase A was purified from the culture filtrate by standard protein purification procedures. The enzyme was characterized as a glycoprotein with a molecular mass of approximately 77 kDa. Both laccase A and laccase B activities were significantly reduced in the hypovirulent (double-stranded RNA-infected) strain UEP1 compared with the isogenic virulent (double-stranded RNA-free) strain EP155/2. PMID- 16349082 TI - Anaerobic Nitrate Respiration by Erwinia carotovora subsp. atroseptica during Potato Tuber Invasion. AB - The in planta induction of anaerobic nitrate respiration by Erwinia carotovora subsp. atroseptica in relation to the in situ oxygen status in soft rotting potato tubers has been investigated. In vitro experiments have shown that nitrate was required for the induction of respiratory nitrate reductase activity in E. carotovora. In addition, oxygen was found to repress this activity. Expression of respiratory nitrate reductase was found in E. carotovora cells extracted from soft rotting potato tuber tissue. However, the rate of nitrite production in these cells was approximately 70-fold lower than the rate recorded in fully induced anaerobic cultures. Oxygen measurements in soft rotting potato tubers indicated that the invading bacteria encounter the lowest oxygen concentration at the interphase between healthy and macerated tissue. Consequently, growth of bacteria present in this specific zone will be stimulated by nitrate which is present in sufficient amounts in tuber tissue. A high nitrate content of the tuber will most likely facilitate the proliferation of E. carotovora in the tuber tissue. PMID- 16349083 TI - Production of Rhizobium Inoculants for Lupinus nootkatensis on Nutrient Supplemented Pumice. AB - The use of the legume Lupinus nootkatensis as a pioneer plant to fight soil erosion and to reclaim eroded soils in Iceland has been under development for a few years. Production of a robust, low-cost bacterial inoculant was therefore a prerequisite for the extended use of this plant. Volcanic pumice is a naturally expanded mineral which is available in vast amounts in Iceland. It was tested as a carrier for solid fermentation of Rhizobium lupini. Nutrient-supplemented pumice containing a small percentage of peat and diatomaceous earth and kept in sterile plastic bags promoted good growth of the bacteria. Viable-colony counts remained stable at 10 to 10/g for at least 35 weeks when the carrier was stored at 22 degrees C. The pumice-based inoculant had good storage and handling properties and could be mixed directly with the seeds during the sowing process. When seeds of L. nootkatensis were sown manually into nutrient-poor eroded sandy soils, about 56% of the first-year plants were successfully nodulated. PMID- 16349084 TI - Aminopeptidase activity in marine chroococcoid cyanobacteria. AB - Synechococci are important primary producers in the ocean and can also utilize some components of the dissolved organic matter (DOM). The readily utilizable DOM in seawater is mainly polymeric (e.g., protein, polysaccharide) or phosphorylated and requires hydrolysis prior to uptake. We examined whether synechococci express ectoenzymes to hydrolyze DOM components and considered the possible significance of ectohydrolases for Synechococcus ecology and organic matter cycling in the sea. Five strains of non-nitrogen-fixing synechococci in axenic cultures were tested for enzyme activities with fluorogenic substrates. All strains show ectocellular aminopeptidase activity, but other enzymes were undetectable. The aminopeptidase level was in the range determined for five marine heterotrophic bacterial isolates tested for comparison. Aminopeptidase was not secreted into the medium; the majority (74%; tested in WH 7803) was cell surface bound, and a small fraction was periplasmic. The periplasmic activity was not released by cold osmotic shock of WH 7803. Phenylmethylsulfonyl fluoride and EDTA, inhibitors of serine and metalloproteases, strongly or completely inhibited WH 7803 aminopeptidase. The enzyme seemed constitutive; per-cell activity did not change during incubations in unenriched seawater, bovine serum albumin, or nitrate replete mineral medium. In natural planktonic assemblages in the Southern California Bight, aminopeptidase activity was correlated with Synechococcus abundance as well as the abundance of other bacteria. Ectocellular aminopeptidase may be common in marine synechococci and play roles in their nitrogen nutrition, particularly in low-nitrate and low-light environments. Since synechococci are much less abundant than heterotrophic bacteria in seawater, the impact of Synechococcus aminopeptidase on proteolysis in the sea is likely to be episodic and restricted to specialized microenvironments. PMID- 16349085 TI - Temporally Regulated Transcriptional Expression of the Genomes of Lactococcal Bacteriophages c2 and sk1. AB - Transcription maps of the Lactococcus lactis subsp. lactis prolate phage c2 and small isometric phage sk1 were constructed. Early and late transcripts were demonstrated in phage c2. Early transcription was localized to within a 7.5-kb EcoRV fragment, and late transcription included the region which encodes the phage structural proteins and a lysin gene. Early, middle, and late transcripts were demonstrated in phage sk1. Transcription was confined to an 11.3-kb region defined by the three EcoRV restriction fragments of 6.2, 4.7, and 0.46 kb during the early part of the sk1 life cycle. Middle gene transcripts extended from the EcoRV site (defining the left-hand limit of early gene expression) through the cos site and included the 4.3-kb PvuII-cos fragment. Late transcription was detected over the remainder of the phage genome. These results indicated that gene expression was temporally regulated at the level of transcription in these two lactococcal phages and that two regions of time-dependent transcription exist in phage c2 and three in phage sk1. PMID- 16349086 TI - Inhibition of Ammonia Oxidation in Nitrosomonas europaea by Sulfur Compounds: Thioethers Are Oxidized to Sulfoxides by Ammonia Monooxygenase. AB - Organic sulfur compounds are well-known nitrification inhibitors. The inhibitory effects of dimethylsulfide, dimethyldisulfide, and ethanethiol on ammonia oxidation by Nitrosomonas europaea were examined. Both dimethylsulfide and dimethyldisulfide were weak inhibitors of ammonia oxidation and exhibited inhibitory characteristics typical of substrates for ammonia monooxygenase (AMO). Depletion of dimethylsulfide required O(2) and was prevented with either acetylene or allylthiourea, two inhibitors of AMO. The inhibition of ammonia oxidation by dimethylsulfide was examined in detail. Cell suspensions incubated in the presence of ammonia oxidized dimethylsulfide to dimethyl sulfoxide. Depletion of six other thioethers was also prevented by treating cell suspensions with either allylthiourea or acetylene. The oxidative products of three thioethers were identified as the corresponding sulfoxides. The amount of sulfoxide formed accounted for a majority of the amount of sulfide depleted. By using gas chromatography coupled with mass spectrometry, allylmethylsulfide was shown to be oxidized to allylmethylsulfoxide by N. europaea with the incorporation of a single atom of O derived from O(2) into the sulfide. This result supported our conclusion that a monooxygenase was involved in the oxidation of allylmethylsulfide. The thioethers are concluded to be a new class of substrates for AMO. This is the first report of the oxidation of the sulfur atom by AMO in whole cells of N. europaea. The ability of N. europaea to oxidize dimethylsulfide is not unique among the ammonia-oxidizing bacteria. Nitrosococcus oceanus, a marine nitrifier, was also demonstrated to oxidize dimethylsulfide to dimethyl sulfoxide. PMID- 16349087 TI - Mechanism-Based Inactivation of Ammonia Monooxygenase in Nitrosomonas europaea by Allylsulfide. AB - Allylsulfide caused an irreversible inactivation of ammonia monooxygenase (AMO) activity (ammonia-dependent O(2) uptake) in Nitrosomonas europaea. The hydroxylamine oxidoreductase activity (hydrazine-dependent O(2) uptake) of cells was unaffected by allylsulfide. Anaerobic conditions or the presence of allylthiourea, a reversible noncompetitive AMO inhibitor, protected AMO from inactivation by allylsulfide. Ammonia did not protect AMO from inactivation by allylsulfide but instead increased the rate of inactivation. The inactivation of AMO followed pseudo-first-order kinetics, but the observed rates did not saturate with increasing allylsulfide concentrations. The time course of recovery of AMO dependent nitrite production after complete inactivation by allylsulfide required de novo protein synthesis. Incubation of cells with allylsulfide prevented the C label from C(2)H(2) (a suicide mechanism-based inactivator of AMO) from being incorporated into the 27-kDa polypeptide of AMO. Some compounds structurally related to allylsulfide were unable to inactivate AMO. We conclude that allylsulfide is a specific, mechanism-based inactivator of AMO in N. europaea. PMID- 16349088 TI - Isolation and Molecular Characterization of Five Marine Cyanophages Propagated on Synechococcus sp. Strain WH7803. AB - Five marine cyanophages propagated on Synechococcus sp. strain WH7803 were isolated from three different oceanographic provinces during the months of August and September 1992: coastal water from the Sargasso Sea, Bermuda; Woods Hole harbor, Woods Hole, Mass.; and coastal water from the English Channel, off Plymouth Sound, United Kingdom. The five cyanophage isolates were found to belong to two families, Myoviridae and Styloviridae, on the basis of their morphology observed in the transmission electron microscope. DNA purified from each of the cyanophage isolates was restricted with a selection of restriction endonucleases, and three distinguishably different patterns were observed. DNA isolated from Myoviridae isolates from Bermuda and the English Channel had highly related restriction patterns, as did DNA isolated from Styloviridae isolates from Bermuda and the English Channel. DNA isolated from the Myoviridae isolate from Woods Hole had a unique restriction pattern. The genome size for each of the Myoviridae isolates was ca. 80 to 85 kb, and it was ca. 90 to 100 kb for each of the Styloviridae isolates. Southern blotting analysis revealed that there was a limited degree of homology among all cyanophage DNAs probed, but clear differences were observed between cyanophage DNA from the Myoviridae and that from the Styloviridae isolates. Polypeptide analysis revealed a clear difference between Myoviridae and Styloviridae polypeptide profiles, although the major, presumably structural, protein in each case was ca. 53 to 54 kDa. PMID- 16349089 TI - Electrochemical prevention of marine biofouling with a carbon-chloroprene sheet. AB - A carbon-chloroprene sheet (CCS) electrode was used for the electrochemical disinfection of the marine gram-negative bacterium Vibrio alginolyticus. When the electrode was incubated in seawater containing 10 cells per ml for 90 min, the amount of adsorbed cells was 4.5 x 10 cells per cm. When a potential of 1.2 V versus a saturated calomel electrode was applied to the CCS for 20 min, 67% of adsorbed cells were killed. This disinfection was due to the direct electrochemical oxidation of cells and not to a change in pH or to the generation of toxic substances, such as chlorine. In a 1-year field experiment, marine biofouling of a CCS-coated cooling pipe caused by attachment of bacteria and invertebrates was considerably reduced by application of a potential of 1.2 V versus a saturated calomel electrode. Since this method requires low potential electrical energy, use of a CCS coating appears to be a suitable method for the clean prevention of marine biofouling. PMID- 16349090 TI - Isolation and Characterization of Nisin-Resistant Leuconostoc mesenteroides for Use in Cabbage Fermentations. AB - Leuconostoc mesenteroides strains that are resistant to high levels of nisin (up to 25,000 IU/ml in broth) were isolated. These nisin-resistant mutants were evaluated to determine their potential use as starter culture strains for cabbage fermentations. We found that some L. mesenteroides strains could be adapted to high levels of nisin resistance, while others could not. The nisin resistance trait was found to be stable for at least 35 generations, in the absence of nisin selection, for all mutants tested. The effects of nisin and salt, separately and in combination, on growth kinetics of the nisin-resistant strains were determined. Salt was the most influential factor on the specific growth rates of the mutants, and no synergistic effect between nisin and salt on specific growth rates was observed. The nisin-resistant strains were unimpaired in their ability to rapidly produce normal heterolactic fermentation end products. The use of these L. mesenteroides mutants as starter cultures in combination with nisin may extend the heterolactic phase of cabbage fermentations. PMID- 16349091 TI - Heterogeneous Distribution of Microbial Activity in Methanogenic Aggregates: pH and Glucose Microprofiles. AB - Methanogenic aggregates, harvested from an upflow anaerobic sludge blanket reactor treating potato starch wastewater, were acclimatized to either glucose or a mixture of sugars and organic nitrogen compounds (i.e., diluted molasses). Both types of granules exhibited internal pH and substrate concentration gradients in mineral medium (pH 7.0, 30 degrees C) as was measured with microelectrodes. Glucose-acclimatized granules suspended in a mineral medium lacking glucose exhibited a distinct internal pH decrease of about 1 U within the granule, suggesting strong metabolism by the acidogenic bacteria. Molasses-acclimatized and aged granules suspended in mineral medium did not exhibit such a pH decrease, suggesting the importance of the metabolic state of these acidogens. The pH gradient did not occur in deactivated granules and was not observable in strongly buffered media (mineral medium containing 33 mM phosphate or reactor liquid). When glucose (0.5 to 5.0 mM) was added to the mineral medium, granules exhibited a convex pH profile. Glucose consumption was located exclusively in the outer 200 to 300 mum of the aggregates (mean diameter = 1.5 mm). The addition of 20 mM 2 bromoethanesulfonic acid to the mineral medium indicated that the higher pH levels in the centre of the granule appeared to be related to the activity of methanogens. It is suggested that acidogenic activity occurs predominantly in the outer 200 to 300 mum of the aggregate and methanogenic activity occurs predominantly in the center of the investigated granules. PMID- 16349092 TI - Disaggregation of Methanosarcina spp. and Growth as Single Cells at Elevated Osmolarity. AB - The effect of medium osmolarity on the morphology and growth of Methanosarcina barkeri, Methanosarcina thermophila, Methanosarcina mazei, Methanosarcina vacuolata, and Methanosarcina acetivorans was examined. Each strain was adapted for growth in NaCl concentrations ranging from 0.05 to 1.0 M. Methanosarcina spp. isolated from both marine and nonmarine sources exhibited similar growth characteristics at all NaCl concentrations tested, demonstrating that these species are capable of adapting to a similar range of medium osmolarities. Concomitant with the adaptation in 0.4 to 1.0 M NaCl, all strains disaggregated and grew as single cells rather than in the characteristic multicellular aggregates. Aggregated cells had a methanochondroitin outer layer, while disaggregated single cells lacked the outer layer but retained the protein S layer adjacent to the cell membrane. Synthesis of glucuronic acid, a major component of methanochondroitin, was reduced 20-fold in the single-cell form of M. barkeri when compared with synthesis in aggregated cells. Strains with the methanochondroitin outer cell layer exhibited enhanced stability at low (<0.2 M NaCl) osmolarity and grew at higher temperatures. Disaggregated cells could be converted back to aggregated cells by gradually readapting cultures to lower NaCl (<0.2 M) and Mg (<0.005 M) concentrations. Disaggregated Methanosarcina spp. could also be colonized and replica plated with greater than 95% recovery rates on solidified agar basal medium that contained 0.4 to 0.6 M NaCl and either trimethylamine, methanol, or acetate as the substrate. The ability to disaggregate and grow Methanosarcina spp. as viable, detergent-sensitive, single cells on agar medium makes these species amenable to mutant selection and screening for genetic studies and enables cells to be gently lysed for the isolation of intact genetic material. PMID- 16349093 TI - Isolation and characterization of actinomycete antagonists of a fungal root pathogen. AB - By use of selective media, 267 actinomycete strains were isolated from four rhizosphere-associated and four non-rhizosphere-associated British soils. Organic media with low nutrient concentrations were found to be best for isolating diverse actinomycetes while avoiding contamination and overgrowth of isolation media by eubacteria and fungi. While all isolates grew well at pHs 6.5 to 8.0, a few were unable to grow at pH 6.0 and a significant number failed to grow at pH 5.5. Eighty-two selected isolates were screened for in vitro antagonism towards Pythium ultimum by use of a Difco cornmeal agar assay procedure. Five isolates were very strong antagonists of the fungus, four were strong antagonists, and ten others were weakly antagonistic. The remaining isolates showed no antagonism by this assay. Additional studies showed that several of the P. ultimum antagonists also strongly inhibited growth of other root-pathogenic fungi. Twelve isolates showing antifungal activity in the in vitro assay were also tested for their effects on the germination and short-term growth of lettuce plants in glasshouse pot studies in the absence of pathogens. None of the actinomycetes prevented seed germination, although half of the isolates retarded seed germination and outgrowth of the plants by 1 to 3 days. During 18-day growth experiments, biomass yields of some actinomycete-inoculated plants were reduced in comparison with untreated control plants, although all plants appeared healthy and well rooted. None of the actinomycetes significantly enhanced plant growth over these short term experiments. For some, but not all, actinomycetes, some correlations between delayed seed germination and reduced 18-day plant biomass yields were seen. For others, plant biomass yields were not reduced despite an actinomycete-associated delay in seed germination and plant outgrowth. Preliminary glasshouse experiments indicated that some of the actinomycetes protect germinating lettuce seeds against damping-off caused by P. ultimum. PMID- 16349094 TI - Bacterioplankton Growth Yield: Seasonal Variations and Coupling to Substrate Lability and beta-Glucosidase Activity. AB - The seasonal variation in the carbon growth yield of pelagic bacteria in the eutrophic lake Frederiksborg Slotsso was studied. The growth yield was determined in dilution culture experiments, in which a substrate of dissolved organic carbon (DOC) from the lake was incubated with a natural bacterioplankton assemblage. Bacterial growth efficiency varied annually from 8 to 60% with an average (and standard deviation) of 41 +/- 11% (n = 29). Simultaneous measurements of growth yield, substrate lability (DOC(L)), chlorophyll and bacterial production, abundance, and extracellular enzymatic activity revealed new aspects of the regulation of bacterial DOC utilization. Growth yield correlated positively to DOC(L) and negatively to beta-d-glucosidase activity. These results indicated a close coupling between the substrate conditions and the physiological response of the bacteria. The large variations in yield within a few days and the close coupling to substrate availability showed that one single global carbon yield factor cannot be expected to apply in pelagic systems. PMID- 16349095 TI - Expression of cryIVA and cryIVB Genes, Independently or in Combination, in a Crystal-Negative Strain of Bacillus thuringiensis subsp. israelensis. AB - The cryIVA and cryIVB genes, encoding the 125- and 135-kDa proteins, respectively, of Bacillus thuringiensis subsp. israelensis, were cloned either alone or together into a shuttle vector and expressed in a nontoxic strain of B. thuringiensis subsp. israelensis. The CryIVB protein was produced at a high level during sporulation and accumulated as inclusions; in contrast, the CryIVA polypeptide did not form such structures unless it was cloned on a higher-copy number plasmid. Transcriptional fusions between the cryIVA or cryIVB gene promoter and the lacZ gene were constructed. The poor synthesis of CryIVA was not due to a poor efficiency of transcription from the cryIVA gene promoter. Mosquitocidal assays performed with purified inclusions showed that CryIVA was toxic for larvae of the species Aedes aegypti, Anopheles stephensi, and Culex pipiens, whereas CryIVB displayed activity only toward Aedes aegypti and Anopheles stephensi. The activity of inclusions containing both polypeptides was higher than that of single-peptide inclusions but was not as high as that of the native crystals, which contain at least four polypeptides. PMID- 16349096 TI - Presence of tetrodotoxin and tetrodotoxin-producing bacteria in freshwater sediments. AB - The occurrence of tetrodotoxin (TTX) in sediment from Lake Suwa, Japan, was confirmed by a tissue culture assay, high-performance liquid chromatography, and gas chromatography-mass spectrometry. Altogether, 17 TTX-producing bacteria belonging to five genera were isolated from Lake Suwa and Pond Inokasira. Our results indicate that TTX and TTX-producing bacteria occur even in freshwater environments. PMID- 16349097 TI - Effect of Medium Composition on the Denitrification of Nitrate by Paracoccus denitrificans. AB - The course of denitrification of nitrate in static cultures of Paracoccus denitrificans was studied. Reduction of nitrate to gaseous nitrogen without accumulation of nitrite because of parallel and balanced activities of nitrate and nitrite reductases was observed in nutrient broth. In minimal liquid cultures supplemented with either methanol, acetate, or ethanol as a sole carbon source, substantial amounts of nitrite (up to 70%) accumulated. The reduction in nitrite concentration began just after the transformation of nitrate to nitrite was completed. The addition of some growth factors to minimal media shortened the bacterial biomass doubling time. A correlation coefficient of 0.71 between the doubling time and the amount of accumulated nitrite in cultures was found. My results indicated that the type of denitrification carried out by P. denitrificans is not stable and depends on the nutritional composition of the culture medium. PMID- 16349098 TI - Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Protein Banding Patterns among Rhizobium leguminosarum biovar phaseoli Strains Isolated from the Mexican Bean Phaseolus coccineus. AB - Several rhizobial strains were isolated from Phaseolus coccineus root nodules and were determined to be Rhizobium leguminosarum biovar phaseoli strains after reinfection of the same host plant. These strains were characterized by cultural procedures (growth on different carbon sources and intrinsic antibiotic resistance) and electrophoretic procedures (sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total proteins). Our results showed that these rhizobia are very similar to each other, especially in their electrophoretic protein banding patterns, suggesting that they might belong to isolated populations. PMID- 16349099 TI - Comparison of Leuconostoc oenos Strains by Pulsed-Field Gel Electrophoresis. AB - Pulsed-field gel electrophoresis of chromosomal DNA digested with NotI or SfiI was used to differentiate individual strains of Leuconostoc oenos. L. oenos isolates with 13 different restriction digest patterns were detected in New Zealand wines undergoing malolactic fermentation. The average genome size was estimated to be 1,800 kb. PMID- 16349100 TI - Physicochemical cell surface and adhesive properties of coryneform bacteria related to the presence and chain length of mycolic acids. AB - The presence and chain length of mycolic acids of bacteria of the genera Corynebacterium, Rhodococcus, Gordona, Mycobacterium, and Arthrobacter and of coryneform bacteria containing a type B peptidoglycan were related to the cell surface hydrophobicity of the bacteria, which in turn was related to adhesion of the cells to defined surfaces such as Teflon and glass. The origin of the overall negative charge of these bacteria is discussed. PMID- 16349101 TI - Measurements of Seasonal Rates and Annual Budgets of Organic Carbon Fluxes in an Antarctic Coastal Environment at Signy Island, South Orkney Islands, Suggest a Broad Balance between Production and Decomposition. AB - We report here the first comprehensive seasonal study of benthic microbial activity in an Antarctic coastal environment. Measurements were made from December 1990 to February 1992 of oxygen uptake and sulfate reduction by inshore coastal sediments at Signy Island, South Orkney Islands, Antarctica. From these measurements the rate of benthic mineralization of organic matter was calculated. In addition, both the deposition rate of organic matter to the bottom sediment and the organic carbon content of the bottom sediment were measured during the same period. Organic matter input to the sediment was small under winter ice cover, and the benthic respiratory activity and the organic content of the surface sediment declined during this period as available organic matter was depleted. On an annual basis, about 32% of benthic organic matter mineralization was anoxic, but the proportion of anoxic compared with oxic mineralization increased during the winter as organic matter was increasingly buried by the amphipod infauna. Fresh organic input occurred as the sea ice melted and ice algae biomass sedimented onto the bottom, and input was sustained during the spring after ice breakup by continued primary production in the water column. The benthic respiratory rate and benthic organic matter content correspondingly increased towards the end of winter with the input of this fresh organic matter. The rates of oxygen uptake during the southern summer (80 to 90 mmol of O(2) m day) were as high as those reported for other sediments at much higher environmental temperatures, and the annual mineralization of organic matter was equally high (12 mol of C m year). Seasonal variations of benthic activity in this antarctic coastal sediment were regulated by the input and availability of organic matter and not by seasonal water temperature, which was relatively constant at between -1.8 and 0.5 degrees C. We conclude that despite the low environmental temperature, organic matter degradation broadly balanced organic matter production, although there may be significant interrannual variations in the sources of the organic matter inputs. PMID- 16349102 TI - Lipase and Esterase Activities of Propionibacterium freudenreichii subsp. freudenreichii. AB - The lipase and esterase activities of eight strains of dairy Propionibacterium freudenreichii subsp. freudenreichii were studied. A lipase activity was detected on whole cells and in the culture supernatant. The highest activity was expressed at 45 degrees C and pH 6.8. An esterase activity was also detected in the culture medium. The electrophoresis of the intracellular fractions of the cells revealed from three to six different esterase activities. Two esterases were common to all the strains. The substrate specificity was dependent on each esterase, but no activity was revealed, in our experimental conditions, on ester substrates with a chain length longer than that of butyrate. PMID- 16349103 TI - Decolorization of Azo, Triphenyl Methane, Heterocyclic, and Polymeric Dyes by Lignin Peroxidase Isoenzymes from Phanerochaete chrysosporium. AB - The ligninolytic enzyme system of Phanerochaete chrysosporium decolorizes several recalcitrant dyes. Three isolated lignin peroxidase isoenzymes (LiP 4.65, LiP 4.15, and LiP 3.85) were compared as decolorizers with the crude enzyme system from the culture medium. LiP 4.65 (H2), LiP 4.15 (H7), and LiP 3.85 (H8) were purified by chromatofocusing, and their kinetic parameters were found to be similar. Ten different types of dyes, including azo, triphenyl methane, heterocyclic, and polymeric dyes, were treated by the crude enzyme preparation. Most of the dyes lost over 75% of their color; only Congo red, Poly R-478, and Poly T-128 were decolorized less than the others, 54, 46, and 48%, respectively. Five different dyes were tested for decolorization by the three purified isoenzymes. The ability of the isoenzymes to decolorize the dyes in the presence of veratryl alcohol was generally comparable to that of the crude enzyme preparation, suggesting that lignin peroxidase plays a major role in the decolorization and that manganese peroxidase is not required to start the degradation of these dyes. In the absence of veratryl alcohol, the decolorization activity of the isoenzymes was in most cases dramatically reduced. However, LiP 3.85 was still able to decolorize 20% of methylene blue and methyl orange and as much as 60% of toluidine blue O, suggesting that at least some dyes can function as substrates for isoenzyme LiP 3.85 but not to the same extent for LiP 4.15 or LiP 4.65. Thus, the isoenzymes have different specificities towards dyes as substrates. PMID- 16349104 TI - Stimulation of Ligninolytic Peroxidase Activity by Nitrogen Nutrients in the White Rot Fungus Bjerkandera sp. Strain BOS55. AB - Bjerkandera sp. strain BOS55, a newly isolated wild-type white rot fungus, produced lignin peroxidase (LiP) in nitrogen (N)-sufficient glucose-peptone medium, whereas no LiP was detectable in N-limited medium. The production of LiP was induced by the peptide-containing components of this medium and also by soy bean protein. Furthermore, the production of manganese-dependent peroxidase was stimulated by organic N sources, although lower production was also evident in N limited medium. Further research showed that the induction of LiP depended on the combination of pH and the type of N source. An amino acid mixture and ammonium induced LiP only at either pH 6 or 7.3, respectively. Peptone induced LiP activity at all pH values tested; however, the highest activity was observed at pH 7.3. The results presented here indicate that Bjerkandera spp. are distinct from the model white rot fungus, Phanerochaete chrysosporium, which produces ligninolytic peroxidases in response to N limitation. PMID- 16349105 TI - Occurrence of killer yeasts in spontaneous wine fermentations from the tuscany region of Italy. AB - The occurrence of killer yeasts in an area of Tuscany (central Italy) was studied. Killer yeasts were found in 88% of spontaneous wine fermentations from 18 wineries. The incidence of killers varied with respect to fermentation stage and vintage period, increasing from the first vintage to successive ones and from the commencement to the end of fermentation. At the end of fermentation, the proportion of killer strains relative to total yeast population was below 25% in 15 cases, above 75% in 6 cases, from 25 to 50% in 5 cases, and from 50 to 75% in 3 cases. Karyotype analysis also showed a mixed killer population in the fermentations in which the killers dominated. PMID- 16349106 TI - Selective Adhesion of Thiobacillus ferrooxidans to Pyrite. AB - Bacterial adhesion to mineral surfaces plays an important role not only in bacterial survival in natural ecosystems, but also in mining industry applications. Selective adhesion was investigated with Thiobacillus ferrooxidans by using four minerals, pyrite, quartz, chalcopyrite, and galena. Escherichia coli was used as a control bacterium. Contact angles were used as indicators of hydrophobicity, which was an important factor in the interaction between minerals and bacteria. The contact angle of E. coli in a 0.5% sodium chloride solution was 31 degrees , and the contact angle of T. ferrooxidans in a pH 2.0 sulfuric acid solution was 23 degrees . E. coli tended to adhere to more hydrophobic minerals by hydrophobic interaction, while T. ferrooxidans selectively adhered to iron containing minerals, such as pyrite and chalcopyrite. Ferrous ion inhibited the selective adhesion of T. ferrooxidans to pyrite competitively, while ferric ion scarcely inhibited such adhesion. When selective adhesion was quenched by ferrous ion completely, adhesion of T. ferrooxidans was controlled by hydrophilic interactions. Adhesion of E. coli to pyrite exhibited a liner relationship on langmuir isotherm plots, but adhesion of T. ferrooxidans did not. T. ferrooxidans recognized the reduced iron in minerals and selectively adhered to pyrite and chalcopyrite by a strong interaction other than the physical interaction. PMID- 16349107 TI - Surface Chemistry of Thiobacillus ferrooxidans Relevant to Adhesion on Mineral Surfaces. AB - Thiobacillus ferrooxidans cells grown on sulfur, pyrite, and chalcopyrite exhibit greater hydrophobicity than ferrous ion-grown cells. The isoelectric points of sulfur-, pyrite-, and chalcopyrite-grown cells were observed to be at a pH higher than that for ferrous ion-grown cells. Microbe-mineral interactions result in change in the surface chemistry of the organism as well as that of the minerals with which it has interacted. Sulfur, pyrite, and chalcopyrite after interaction with T. ferrooxidans exhibited a significant shift in their isoelectric points from the initial values exhibited by uninteracted minerals. With antibodies raised against sulfur-grown T. ferrooxidans, pyrite- and chalcopyrite-grown cells showed immunoreactivity, whereas ferrous ion-grown cells failed to do so. Fourier transform infrared spectroscopy of sulfur-grown cells suggested that a proteinaceous new cell surface appendage synthesized in mineral-grown cells brings about adhesion to the solid mineral substrates. Such an appendage was found to be absent in ferrous ion-grown cells as it is not required during growth in liquid substrates. PMID- 16349108 TI - Characterization of the Binding of Gallium, Platinum, and Uranium to Pseudomonas fluorescens by Small-Angle X-Ray Scattering and Transmission Electron Microscopy. AB - Small-angle X-ray scattering (SAXS) was used to determine the binding of Ga, U, and Pt to Pseudomonas fluorescens in aqueous buffer. Atomic absorption spectrophotometry was used to quantify the heavy metals during bulk analysis, whereas transmission electron microscopy of whole mounts and thin sections was used to determine the locations of the cell-bound metal precipitates, as well as their sizes and physical structures. Energy-dispersive X-ray spectroscopy confirmed the compositions and identities of the precipitates and helped show that they were associated primarily with the envelope layers of the bacteria. Unlike Ga and Pt, which were located only at the cell surface, U was also found intracellularly in approximately 10% of the cells. This cytoplasmic location ultimately killed and lysed the cells. Surface-bound Ga and U were spread over the entire cell envelope (outer membrane-peptidoglycan-plasma membrane complex), whereas Pt was associated only with the lipopolysaccharide-rich, external face of the outer membrane. SAXS confirmed these data and showed that the bacteria were metal-enshrouded particles that were 1.0 to 1.5 mum in diameter. SAXS also provided a statistically significant representation of the bound metal precipitates, which ranged in size from 10 nm to 1 mum. The correlation between the microscopic data and the scattering data was extremely good. Since SAXS is performed in an aqueous milieu, it yields a more representative picture of the physical state of the metal bound to cell surfaces. PMID- 16349109 TI - Distribution of Viruses and Dissolved DNA along a Coastal Trophic Gradient in the Northern Adriatic Sea. AB - The distribution of viral and other microbial abundances as well as the concentrations of dissolved DNA (D-DNA) along a trophic gradient in the northern Adriatic Sea were determined. Virus abundances, covering a range of 1.2 x 10 to 8.7 x 10 liter were on average 2.5-fold higher in eutrophic than in mesotrophic stations. A 2.5-fold enrichment was also measured for chlorophyll a concentrations, whereas the densities of bacteria and heterotrophic nanoflagellates were only approximately 1.5-fold higher. The frequency of bacteria containing mature phage increased linearly with bacterial abundance. Assuming that mature phage is only visible during the last 14 to 27% of the latent period (L. M. Proctor, A. Okubo, and J. A. Fuhrman, Microb. Ecol. 25:161 182, 1993), we estimated that between 3.5 and 7.3% of the bacterial population was infected at mesotrophic stations versus between 7.0 and 19.5% at eutrophic stations, indicating that the bacterial mortality due to viral lysis might increase with the degree of eutrophication. The frequency of bacteria with mature phage and the burst size varied significantly with the bacterial morphotype; rod shape cells, the most abundant morphotype, showed low infection rates but a high burst size. Concentrations of D-DNA varied significantly with season but not with trophic conditions. The estimated percentage of viral DNA on total D-DNA concentrations averaged 17.1% (range, 0.7 to 88.3%). Some kind of interaction between heterotrophic nanoflagellates and viruses is proposed. We conclude (i) that the significance of viruses varies with changing trophic conditions and (ii) that viral activity may play a significant role in food web structure under changing trophic conditions. PMID- 16349110 TI - Impact of Chlorine and Heat on the Survival of Hartmannella vermiformis and Subsequent Growth of Legionella pneumophila. AB - Hartmannella vermiformis, a common amoebal inhabitant of potable-water systems, supports intracellular multiplication of Legionella pneumophila and is probably important in the transportation and amplification of legionellae within these systems. To provide a practical guide for decontamination of potable-water systems, we assessed the chlorine and heat resistance of H. vermiformis. H. vermiformis cysts and trophozoites were treated independently with chlorine at concentrations of 2.0 to 10.0 ppm for 30 min and then cocultured with L. pneumophila. Both cysts and trophozoites were sensitive to concentrations between 2.0 and 4.0 ppm and above (trophozoites somewhat more so than cysts), and 10.0 ppm was lethal to both forms. Hartmannellae treated with chlorine up to a concentration of 4.0 ppm supported the growth of legionellae. To determine whether heat would be an effective addendum to chlorine treatment of amoebae, hartmannellae were subjected to temperatures of 55 and 60 degrees C for 30 min and alternatively to 50 degrees C followed by treatment with chlorine at a concentration of 2 ppm. Fewer than 0.05% of the amoebae survived treatment at 55 degrees C, and there were no survivors at 60 degrees C. Pretreatment at 50 degrees C appeared to make hartmannella cysts more susceptible to chlorine but did not further reduce the concentration of trophozoites. PMID- 16349111 TI - Quantitative reverse sample genome probing of microbial communities and its application to oil field production waters. AB - This paper presents a protocol for quantitative analysis of microbial communities by reverse sample genome probing is presented in which (i) whole community DNA is isolated and labeled in the presence of a known amount of an added internal standard and (ii) the resulting spiked reverse genome probe is hybridized with a master filter on which denatured genomic DNAs from bacterial standards isolated from the target environment were spotted in large amounts (up to 1,500 ng) in order to improve detection sensitivity. This protocol allowed reproducible fingerprinting of the microbial community in oil field production waters at 19 sites from which water and biofilm samples were collected. It appeared that selected sulfate-reducing bacteria were significantly enhanced in biofilms covering the metal surfaces in contact with the production waters. PMID- 16349112 TI - Effect of Manganese on Lignin Degradation by Pleurotus ostreatus during Solid State Fermentation. AB - Lignin degradation by Pleurotus ostreatus was studied under solid-state fermentation (SSF) in chemically defined medium containing various levels of Mn. Degradation of [C]lignin prepared from cotton branches to soluble products, as well as its mineralization to CO(2), was enhanced by the addition of Mn. The effect of malonate on lignin mineralization was most marked during the first 10 days of SSF, in a treatment amended with 73 muM Mn. A high concentration of Mn (4.5 mM) caused inhibition of both fungal growth and mineralization rates during the first 2 weeks of incubation. Addition of malonate reversed this effect because of chelation of Mn. Mn was found to precipitate in all treatments, with or without the addition of malonate. alpha-Keto-gamma-methiolbutyric acid cleavage to ethylene, an indication of OH production, was observed as early as 3 days of incubation in all treatments. PMID- 16349113 TI - Cloning and Mutagenesis of a Cytochrome P-450 Locus from Bradyrhizobium japonicum That Is Expressed Anaerobically and Symbiotically. AB - Cytochromes P-450, which in many organisms participate in the metabolism of a variety of endobiotic and xenobiotic substances, are synthesized by symbiotic bacteroids of Bradyrhizobium japonicum. Polyclonal antibodies were raised against two cytochromes P-450 (CYP112 and CYP114) purified from bacteroids. A lambda gt11 expression clone of B. japonicum USDA 110 DNA that reacted with the anti-CYP112 antibody was obtained and was used to screen a library of USDA 110 genomic DNA in pLAFR1 for a clone of the P-450 locus. Forced expression of subclones of the P 450 locus in Escherichia coli produced polypeptides that reacted with either the anti-CYP112 antibody or the anti-CYP114 antibody; no cross-reactivity was evident. A Western blot (immunoblot) analysis showed that neither protein was present in free-living aerobically grown B. japonicum cells, but that both proteins were present in cells grown anaerobically, as well as in bacteroids. A mutant strain disrupted in the CYP112 locus produced neither CYP112 nor CYP114, indicating that the mutation was polar for CYP114. The mutant produced effective nodules on soybeans, even though the bacteroids contained no detectable P-450. This suggests that the cytochromes P-450 which we examined are not involved in an essential symbiotic function. PMID- 16349114 TI - Outer Membrane Proteins and Lipopolysaccharides in Pathovars of Xanthomonas campestris. AB - Variations in the outer membrane proteins (OMPs) and lipopolysaccharides (LPSs) of 54 isolates belonging to 16 different pathovars of Xanthomonas campestris were characterized. OMP samples prepared by sarcosyl extraction of cell walls and LPS samples prepared by proteinase K treatment of sonicated cells were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of 4 M urea. In general, the OMP and LPS profiles within each pathovar were very similar but different from the profiles of other pathovars. Heterogeneity in OMP and LPS profiles was observed within X. campestris pv. campestris, X. campestris pv. translucens, and X. campestris pv. vesicatoria. LPSs were isolated from six X. campestris pathovars, which fell into two major groups on the basis of O antigenicity. The O antigens of X. campestris pv. begoniae, X. campestris pv. graminis, and X. campestris pv. translucens cross-reacted with each other; the other group consisted of X. campestris pv. campestris, X. campestris pv. pelargonii, and X. campestris pv. vesicatoria. A chemical analysis revealed a significant difference between the compositions of the neutral sugars of the LPSs of those two groups; the LPSs of the first group contained xylose and a 6-deoxy-3 O-methyl hexose, whereas the LPSs of the other group lacked both sugars. PMID- 16349115 TI - Uptake Hydrogenase (Hup) in Common Bean (Phaseolus vulgaris) Symbioses. AB - Strains of Rhizobium forming nitrogen-fixing symbioses with common bean were systematically examined for the presence of the uptake hydrogenase (hup) structural genes and expression of uptake hydrogenase (Hup) activity. DNA with homology to the hup structural genes of Bradyrhizobium japonicum was present in 100 of 248 strains examined. EcoRI fragments with molecular sizes of approximately 20.0 and 2.2 kb hybridized with an internal SacI fragment, which contains part of both bradyrhizobial hup structural genes. The DNA with homology to the hup genes was located on pSym of one of the bean rhizobia. Hup activity was observed in bean symbioses with 13 of 30 strains containing DNA homologous with the hup structural genes. However, the Hup activity was not sufficient to eliminate hydrogen evolution from the nodules. Varying the host plant with two of the Hup strains indicated that expression of Hup activity was host regulated, as has been reported with soybean, pea, and cowpea strains. PMID- 16349116 TI - Thermal Inactivation of Aeromonas hydrophila As Affected by Sodium Chloride and Ascorbic Acid. AB - The combined effects of sodium chloride (0, 1.0, 1.5, and 3.0%) and ascorbic acid (0, 1.0, and 2.0 mmol/liter) with mild heat (46 degrees C) on the survival of Aeromonas hydrophila were evaluated. Because of the nonlinear nature of the survivor curves obtained, several equations yielding an R (coefficient of multiple determination) of congruent with 1 were tested. The equation that most closely fit the curvature of the observed data set was a hyperbolic function. Equation coefficients were combined to obtain a so-called death value. This value (46.67% explained variance) was calculated by extracting the larger eigenvalue and the relative eigenvector from the correlation matrix of the coefficients. the effects of the experimental factors on the death value were described by a quadratic response surface model. Results revealed that the death value was not influenced by the presence of ascorbic acid. However, increased mortality resulted from the interaction between sodium chloride and ascorbic acid. PMID- 16349117 TI - Effect of organic matter decomposition level on bacterial species diversity and composition in relationship to pythium damping-off severity. AB - Rhizosphere bacteria were isolated from root tip segments of cucumber seedlings grown in a suppressive, slightly decomposed light-colored peat mix, a conducive, more decomposed dark-colored peat mix, and a suppressive dark peat mix amended with composted hardwood bark. The bacteria were identified by a gas chromatographic fatty acid methyl ester analysis. The total number of taxa recovered from a single root tip segment ranged from 9 to 18. No single taxon predominated on all root tip segments harvested from any of the mixes. The highest relative population density reached by a given taxon on any root tip segment was 45%. Hill's first and second diversity numbers, the modified Hill's ratio, and Hurlbert's rarefaction method, which were used as measures of species diversity, indicated that the organic matter decomposition level of the potting mixes did not affect bacterial species diversity. Bray-Curtis polar ordination and Dice resemblance functions, however, indicated that the organic matter decomposition level of a mix significantly influenced the composition of bacterial species in the rhizosphere. Pseudomonas spp. and other taxa capable of inducing suppression of pythium damping-off predominated in the suppressive mixes. These organisms were absent from the conducive mix, in which Arthrobacter and Bacillus spp. predominated. Although effective bacterial biocontrol agents were isolated from both the suppressive mixes and the conducive mix, the majority were isolated from the less decomposed suppressive mixes. Finally, the efficacy of strains was significantly greater in the slightly decomposed light peat mix than in the decomposed dark peat mix. Natural disease suppression within these mixes was associated with the organic matter decomposition level and the bacterial species compositions of the mixes. PMID- 16349118 TI - Cloning and Characterization of Aerobactin Biosynthesis Genes of the Biological Control Agent Enterobacter cloacae. AB - Five strains of Enterobacter cloacae that are biological control agents of Pythium damping-off diseases produced the hydroxamate siderophore aerobactin under iron-limiting conditions. Genes determining aerobactin biosynthesis of the biocontrol strain E. cloacae EcCT-501 were localized to a 12.3-kb region, which conferred aerobactin production to Escherichia coli DH5alpha. The aerobactin biosynthesis genes of E. cloacae hybridized to those of the pColV-K30 plasmid of E. coli, but restriction patterns of the aerobactin regions of pColV-K30 and E. cloacae differed. A derivative strain with a deletion in the aerobactin biosynthesis locus was as effective as strain EcCT-501 in biological control of Pythium damping-off of cucumber. Thus, aerobactin production did not contribute significantly to the biological control activity of EcCT-501 under the conditions of this study. PMID- 16349119 TI - Isolation of a Degeneration-Resistant Mutant of Clostridium acetobutylicum NCIMB 8052. AB - Unless periodically grown from germinated spores, Clostridium acetobutylicum tends to degenerate (that is, to spontaneously lose the capacity both to produce solvents and to develop into spores). To obtain mutants that are deficient in degeneration, C. acetobutylicum NCIMB 8052 was mated with Enterococcus faecalis BM4110 harboring transposon Tn1545. We developed a degeneration resistance assay based on a secondary effect of degeneration, the production of toxic levels of acetic and butyric acids. Erythromycin-resistant transconjugant clones were tested individually for longevity by repeated and timely subculturing. One long lived mutant, A10, survived 18 +/- 3 transfers (mean +/- standard deviation; n = 20) before extinction, while the wild type (parental cells) survived 6.6 +/- 1.5 transfers (n = 11). The three-fold difference in longevity is statistically significant. In a batch culture in a rich medium, the wild-type cells degenerated within 24 h after inoculation with 1% of an overnight culture derived from germinated spores. In contrast, A10 cells were able to switch to solventogenesis and to sporulate. In a minimal medium with greater buffering capacity, both cell types produced solvents and spores. Southern blots of EcoRI and HindIII restriction digests of A10 chromosomal DNA (but not parental DNA) showed that only one copy of Tn1545 was inserted into the clostridial chromosome. Our findings are consistent with the hypothesis that there was an alteration at a regulatory locus that was effected by the insertion of the transposon. PMID- 16349120 TI - Growth and Energy Generation by Lactococcus lactis subsp. lactis biovar diacetylactis during Citrate Metabolism. AB - Growth of Lactococcus lactis subsp. lactis biovar diacetylactis was observed on media with citrate as the only energy source. At pH 5.6, steady state was achieved in a chemostat on a citrate-containing medium in the absence of a carbohydrate. Under these conditions, pyruvate, acetate, and some acetoin and butanediol were the main fermentation products. This indicated that energy was conserved in L. lactis subsp. lactis biovar diacetylactis during citrate metabolism and presumably during the conversion of citrate into pyruvate. The presumed energy-conserving step, decarboxylation of oxaloacetate, was studied in detail. Oxaloacetate decarboxylase was purified to homogeneity and characterized. The enzyme has a native molecular mass of approximately 300 kDa and consists of three subunits of 52, 34, and 12 kDa. The enzyme is apparently not sodium dependent and does not contain a biotin moiety, and it seems to be different from the energy-generating oxaloacetate decarboxylase from Klebsiella pneumoniae. Energy-depleted L. lactis subsp. lactis biovar diacetylactis cells generated a membrane potential and a pH gradient immediately upon addition of citrate, whereas ATP formation was slow and limited. In contrast, lactose energization resulted in rapid ATP formation and gradual generation of a proton motive force. These data were confirmed during studies on amino acid uptake. alpha Aminoisobutyrate uptake was rapid but glutamate uptake was slow in citrate energized cells, whereas lactose-energized cells showed the reverse tendency. These data suggest that, in L. lactis subsp. lactis bv. diacetylactis, a proton motive force could be generated during citrate metabolism as a result of electrogenic citrate uptake or citrate/product exchange together with proton consumption by the intracellular oxaloacetate decarboxylase. PMID- 16349121 TI - Growth of Azotobacter vinelandii UWD in Fish Peptone Medium and Simplified Extraction of Poly-beta-Hydroxybutyrate. AB - Azotobacter vinelandii UWD was grown in a fermentor with glucose medium with and without 0.1% fish peptone (FP) in batch and fed-batch cultures for the production of the natural bioplastic poly-beta-hydroxybutyrate (PHB). Strain UWD formed PHB five times faster than cell protein during growth in glucose and NH(4), but PHB synthesis stopped when NH(4) was depleted and nitrogen fixation started. When FP was added to the same medium, PHB accumulated 16 times faster than cell protein, which in turn was inhibited by 40%, and PHB synthesis was unaffected by NH(4) depletion. Thus, FP appeared to be used as a nitrogen source by these nitrogen fixing cells, which permitted enhanced PHB synthesis, but it was not a general growth stimulator. The addition of FP to the medium led to the production of large, pleomorphic, osmotically sensitive cells that demonstrated impaired growth and partial lysis, with the leakage of DNA into the culture fluid, but these cells were still able to synthesize PHB at elevated rates and efficiency. When FP was continuously present in fed-batch culture, the yield in grams of polymer per gram of glucose consumed was calculated to range from 0.43 g/g, characteristic of nongrowing cells, to an unprecedented 0.65 g/g. Separation of an FP-free growth phase from an FP-containing growth phase in fed-batch culture resulted in better growth of these pleomorphic cells and good production of PHB (yield, 0.32 g/g). The fragility of these cells was exploited in a simple procedure for the extraction of high-molecular-weight PHB. The cells were treated with 1 N aqueous NH(3) (pH 11.4) at 45 degrees C for 10 min. This treatment removed about 10% of the non-PHB mass from the pellet, of which 60 to 77% was protein. The final product consisted of 94% PHB, 2% protein, and 4% nonprotein residual mass. The polymer molecular weight (1.7 x 10 to 2.0 x 10) and dispersity (1.0 to 1.9) were not significantly affected (P = 0.05) by this treatment. In addition, the NH(3) extraction waste could be recycled in the fermentation as a nitrogen source, but it did not promote PHB production like FP. A scheme for improved downstream extraction of PHB as well as the merits of using pleomorphic cells in the production of bioplastics is discussed. PMID- 16349122 TI - Analysis of Fungal Pellets by UV-Visible Spectrum Diffuse Reflectance Spectroscopy. AB - The application of the UV-visible spectrum diffuse reflectance spectroscopy for the determination of intracellular pH in vivo, for determination of cytochrome content, and for the noninvasive in vivo detection of the redox state of fungal mitochondrial cytochromes in filamentous fungi is introduced. The time course of the intracellular pH values, mitochondrial cytochromes, and CO-binding pigments content and the correlations between the actual redox state of cytochrome aa(3) and saturation of growth medium with oxygen in pellets of the basidiomycete Phanerochaete chrysosporium were determined. As the test microorganism, the yeast Saccharomyces cerevisiae was used. UV-visible spectrum diffuse reflectance spectroscopy proved to be a promising method for the quick and simple analysis of light-impermeable biological structures for which the classical transmittance spectrophotometric methods are difficult to implement. PMID- 16349123 TI - Microbial delignification with white rot fungi improves forage digestibility. AB - Three wild-type white rot fungi and two cellulase-less mutants developed from Phanerochaete chrysosporium K-3 (formerly Sporotrichum pulverulentum) were tested for their ability to delignify grass cell walls and improve biodegradation by rumen microorganisms. Fungal-treated and control stems of Bermuda grass were analyzed for their content of ester- and ether-linked aromatics by using alkali extraction and gas chromatography, for in vitro dry weight digestion and production of volatile fatty acids in in vitro fermentations with mixed ruminal microorganisms, for loss of lignin and other aromatics from specific cell wall types by using microspectrophotometry, and for structural changes before and after in vitro degradation by rumen microorganisms by using transmission electron microscopy. P. chrysosporium K-3 and Ceriporiopsis subvermispora FP 90031-sp produced the greatest losses in lignin and improved the biodegradation of Bermuda grass over that of untreated control substrate. However, C. subvermispora removed the most lignin and significantly improved biodegradation over all other treatments. Phellinus pini RAB-83-19 and cellulase-less mutants 3113 and 85118 developed from P. chrysosporium K-3 did not improve the biodegradation of Bermuda grass lignocellulose. Results indicated that C. subvermispora extensively removed ester-linked p-coumaric and ferulic acids and also removed the greatest amount of non-ester-linked aromatics from plant cell walls. Microscopic observations further indicated that C. subvermispora removed esters from parenchyma cell walls as well as esters and lignin from the more recalcitrant cell walls (i.e., sclerenchyma and vascular tissues). C. subvermispora improved in vitro digestion and volatile fatty acid production by ruminal microorganisms by about 80%, while dry matter loss due to fungi was about 20% greater than loss in untreated control stems. The chemical and structural studies used identified sites of specific fungal attack and suggested mechanisms whereby improvement occurred. PMID- 16349124 TI - Identification of Molecular Variants in Mitochondrial DNAs of Members of the Genera Beauveria, Verticillium, Paecilomyces, Tolypocladium, and Metarhizium. AB - A set of five mitochondrial (mt) probes derived from a strain of Beauveria bassiana was used to evaluate the similarity of mtDNAs from 15 additional isolates of this fungus and five genera of other entomopathogenic fungi. The probes and genes encoded for (shown in parentheses) were pBbmtE2 (NADI, ATP6), pBbmtE3 (ATP6, small rRNA [srRNA]), pBbmtE4 (srRNA, CO3, NAD6), pBbSE1 (NAD6, tRNA, large rRNA [lrRNA]), and pBbXS1 (lrRNA). The probes produced identical hybridization patterns in EcoRI-digested DNA from nearly all isolates of B. bassiana and Beauveria caledonica. Similar patterns were also observed with Beauveria densa. The isolates of B. caledonica and B. densa DNAs could be differentiated from each other and from B. bassiana on the basis of a HindIII digestion and probing with pBbmtE3. Probe pBbmtE2 produced either a 5.0-kb or a 4.1-kb band in all of the B. bassiana isolates. This observation was used to categorize the mtDNA of B. bassiana into two types, designated A and B. Hybridization of the five probes produced distinct banding patterns in Beauveria brongniartii, Tolypocladium cylindrosporum, Tolypocladium nivea, Metarhizium anisopliae, Verticillium lecanii, and Paecilomyces farinosus. Hybridizations carried out with multiple probes simultaneously present produced unique patterns which characterized the B. bassiana group from all other fungi tested. These results are discussed in terms of how mtDNA polymorphisms in B. bassiana may relate to natural population structures, mt transmission in deuteromycetes, and the use of mtDNA polymorphisms in structural analysis of mtDNA. PMID- 16349126 TI - A Novel Delta5-Desaturase-Defective Mutant of Mortierella alpina 1S-4 and Its Dihomo-gamma-Linolenic Acid Productivity. AB - A novel Delta5-desaturase-defective mutant was derived from an arachidonic acid producing fungus, Mortierella alpina 1S-4, after treating the parental spores with N-methyl-N'-nitro-N-nitrosoguanidine. The mutant produced only a trace (about 1%) amount of arachidonic acid, and the ratio of dihomo-gamma-linolenic acid (DGLA) to total fatty acids in each lipid class was markedly high, accounting for as much as 60% in phosphatidylcholine. Under submerged batch culture conditions, the mutant produced 2.4 g of DGLA per liter (43.3% of total fatty acids) when grown at 28 degrees C for 7 days in a 5-liter jar fermentor. The other major (more that 1%) fatty acids were palmitic acid (21.2%), stearic acid (9.6%), oleic acid (14.3%), linoleic acid (4.4%), and gamma-linolenic acid (5.8%). About 80 mol% of the DGLA produced was found in triacylglycerol. PMID- 16349125 TI - Heat Shock Induction of Manganese Peroxidase Gene Transcription in Phanerochaete chrysosporium. AB - The expression of manganese peroxidase (MnP) in nitrogen-limited cultures of Phanerochaete chrysosporium is regulated by heat shock at the level of gene transcription. Nitrogen limitation and manganous ion [Mn(II)] previously have been shown to regulate mnp gene transcription. Northern (RNA) blot analysis demonstrates that 45 degrees C heat shock results in the accumulation of mnp mRNA, even in cells grown in the absence of Mn. Heat shock induces mnp gene transcription in 4- or 5-day-old cells, and mnp mRNA is detectable after 15 min at 45 degrees C. Maximum accumulation of mnp mRNA is observed 1 to 2 h after transfer of cultures to 45 degrees C. Two hours after heat shock-induced cultures grown in the absence of Mn are transferred back to 37 degrees C, mnp mRNA is no longer detectable. Higher levels of mnp mRNA are obtained with simultaneous induction by Mn and heat shock than by either treatment alone. Neither MnP enzyme activity nor protein is detectable in heat-shocked cultures grown in the absence of Mn. However, higher MnP activity is found in the extracellular medium of cultures induced by both heat shock and Mn than in the medium of cultures induced by Mn alone. These results suggest that the putative heat shock elements found in the promoter region of the mnp genes are physiologically functional and that Mn may be required for a posttranscriptional step of MnP production under heat shock conditions. PMID- 16349127 TI - Hydrophobic and electrostatic cell surface properties of thermophilic dairy streptococci. AB - Microbial adhesion to hydrocarbons (MATH) and microelectrophoresis were done in 10 mM potassium phosphate solutions to characterize the surfaces of thermophilic dairy streptococci, isolated from pasteurizers. Regardless of whether they were grown (in M17 broth) with lactose, sucrose, or glucose added, strains were relatively hydrophilic (showing low initial removal rates by hexadecane) and slightly negatively charged. A tendency exists for cells grown with sucrose added to be more hydrophilic than cells grown with glucose or lactose added. Also, the lowest isoelectric points, i.e., the pH values for which the zeta potentials are zero, were measured for strains with glucose added to the growth medium. The isoelectric points for the strains were all rather high, between pH 3 and 5, indicative of protein-rich surfaces, although X-ray photoelectron spectroscopy did not measure excessively large amounts of nitrogen on the cell surfaces. Both MATH and microelectrophoresis were done as a function of pH. Maxima in hydrophobicity were observed at certain pH values. Usually these pH values were in the range of the isoelectric points of the cells. Thus it appears that MATH measures an interplay of hydrophobicity and electrostatic interactions. MATH measures solely hydrophobicity only when electrostatic interactions are absent, i.e., close to the isoelectric points of the cells. Considering that these thermophilic streptococci are all rather hydrophilic, a possible pathway to prevent fouling in the pasteurization process might be to render the heat exchanger plates of the pasteurizer more hydrophobic. PMID- 16349128 TI - Glutamine Involvement in Nitrogen Control of Gibberellic Acid Production in Gibberella fujikuroi. AB - When the fungus Gibberella fujikuroi ATCC 12616 was grown in fermentor cultures, both intracellular kaurene biosynthetic activities and extracellular GA(3) accumulation reached high levels when exogenous nitrogen was depleted in the culture. Similar patterns were exhibited by several nonrelated enzymatic activities, such as formamidase and urease, suggesting that all are subject to nitrogen regulation. The behavior of the enzymes involved in nitrogen assimilation (glutamine synthetase, glutamate dehydrogenase, and glutamate synthase) during fungal growth in different nitrogen sources suggests that glutamine is the final product of nitrogen assimilation in G. fujikuroi. When ammonium or glutamine was added to hormone-producing cultures, extracellular GA(3) did not accumulate. However, when the conversion of ammonium into glutamine was inhibited by L-methionine-DL-sulfoximine, only glutamine maintained this effect. These results suggest that glutamine may well be the metabolite effector in nitrogen repression of GA(3) synthesis, as well as in other nonrelated enzymatic activities in G. fujikuroi. PMID- 16349129 TI - Remobilization of Heavy Metals Retained as Oxyhydroxides or Silicates by Bacillus subtilis Cells. AB - The stability of heavy metals bound as either silicates or oxyhydroxides by the surfaces of Bacillus subtilis with changes in pH and presence of competing cations and EDTA was studied. Wall-bound silicate was very stable. Of all conditions, a pH of 3.0 (by HNO(3)) promoted the greatest release of Cd (87% [wt/wt]), Pb (77%), or Cu (54%). Bacterial Pb resisted extraction by EDTA, and Cd that was included in bacterial-silicates proved difficult to remobilize. PMID- 16349130 TI - Purification and Characterization of an l-Aminopeptidase from Pseudomonas putida ATCC 12633. AB - An l-aminopeptidase of Pseudomonas putida, used in an industrial process for the hydrolysis of d,l-amino acid amide racemates, was purified to homogeneity. The highly l-enantioselective enzyme resembled thiol reagent-sensitive alkaline serine proteinases and was strongly activated by divalent cations. It possessed a high substrate specificity for dipeptides and alpha-H amino acid amides, e.g., l phenylglycine amide. PMID- 16349131 TI - Influence of packaging and processing conditions on the decontamination of laboratory biomedical waste by steam sterilization. AB - The conditions for optimal steam decontamination of polypropylene bags half loaded with laboratory biomedical waste were studied (276 bags were processed). Controls were single-closed bags without water added or incisions made in the top, standing freely in an autoclave set at 121 degrees C. The average time required to reach 121 degrees C at the load center was 46 min for controls. A significant increase in this time occurred following addition of water to bags without incisions (60 min), with double bagging (60 min), or when using vertical containers (82 min). A significant decrease occurred when bags were slashed (37 min) or processed at 123 degrees C (32 min) or 132 degrees C (19 min). Horizontal containers or addition of water to slashed bags had no significant effect. PMID- 16349132 TI - Isolation and Characterization of a Novel Atrazine Metabolite Produced by the Fungus Pleurotus pulmonarius, 2-Chloro-4-Ethylamino-6-(1-Hydroxyisopropyl)Amino 1,3,5-Triazine. AB - The white rot fungus Pleurotus pulmonarius exhibited metabolism of atrazine (2 chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) in liquid culture, producing the dealkylated metabolites desethylatrazine, desisopropylatrazine, and desethyl desisopropylatrazine. A fourth, unknown metabolite was also produced. It was isolated and was identified as 2-chloro-4-ethylamino-6-(1-hydroxyisopropyl)amino 1,3,5-triazine by gas chromatography-mass spectrometry, Fourier transformed infrared spectroscopy, and H nuclear magnetic resonance analysis. The structure of this metabolite was confirmed by chemical synthesis of the compound and comparison with the fungally produced metabolite. PMID- 16349133 TI - Abiotic reduction of 4-chloronitrobenzene to 4-chloroaniline in a dissimilatory iron-reducing enrichment culture. AB - 4-chloronitrobenzene (4-Cl-NB) was rapidly reduced to 4-chloroaniline with half lives of minutes in a dissimilatory Fe(III)-reducing enrichment culture. The initial pseudo-first-order rate constants at 25 degrees C ranged from 0.11 to 0.19 per minute. The linear Arrhenius correlation in a temperature range of 6 to 85 degrees C and the unchanged reactivity after pasteurization indicated that the nitroreduction occurred abiotically. A fine-grained black solid which was identified as poorly crystalline magnetite (Fe(3)O(4)) by X-ray diffraction accumulated in the enrichments. Magnetite produced by the Fe(III)-reducing bacterium Geobacter metallireducens GS-15 and synthetic magnetite also reduced 4 Cl-NB. These results suggest that the reduction of 4-Cl-NB by the enrichment material was a surface-mediated reaction by dissimilatory formed Fe(II) associated with magnetite. PMID- 16349134 TI - Monitoring of bacterial sugars and hydroxy Fatty acids in dust from air conditioners by gas chromatography-mass spectrometry. AB - Bacterial levels in dust collected from hospital air-conditioning filters were examined by chemical analysis (without prior culture). The dust was analyzed by gas chromatography-mass spectrometry after hydrolysis and derivatization. l Glycero-d-mannoheptose and hydroxy fatty acids (3-OH 12:0 and 3-OH 14:0) (primarily found in lipopolysaccharide) and muramic acid (a chemical marker for bacterial peptidoglycan) were present at higher levels in dust collected from filters primarily contacting outdoor (as opposed to indoor) air. The ratio of l glycero-d-mannoheptose to muramic acid in dust (compared with those of a group of gram-positive and gram-negative bacteria) suggested that both dust types contained appreciable numbers of gram-negative bacteria. There is potential for the chemical assessment of the microbial content of airborne dust. PMID- 16349135 TI - Effects of Preservatives on Viability of Cryptosporidium parvum Oocysts. AB - Potassium dichromate and formalin reduced the viability of Cryptosporidium parvum oocysts as assessed by inclusion or exclusion of 4',6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) and excystation. Some formalin-treated oocysts containing dead sporozoites excluded PI; that this fluorogenic assay relies not solely upon exclusion of PI but also upon highlighting of sporozoite nuclei by DAPI is reiterated. PMID- 16349136 TI - Minimal Requirements for Exponential Growth of Lactococcus lactis. AB - A minimal growth medium containing glucose, acetate, vitamins, and eight amino acids allowed for growth of Lactococcus lactis subsp. lactis, with a specific growth rate in batch culture of mu = 0.3 h. With 19 amino acids added, the growth rate increased to mu = 0.7 h and the exponential growth phase proceeded until high cell concentrations were reached. We show that morpholinepropanesulfonic acid (MOPS) is a suitable buffer for L. lactis and may be applied in high concentrations. PMID- 16349137 TI - Reductive Dehalogenation of Chlorinated Benzenes and Toluenes under Methanogenic Conditions. AB - [This corrects the article on p. 3270 in vol. 59.]. PMID- 16349139 TI - Cloning and Sequence Analysis of the meso-Diaminopimelate Decarboxylase Gene from Bacillus methanolicus MGA3 and Comparison to Other Decarboxylase Genes. AB - [This corrects the article on p. 2935 in vol. 59.]. PMID- 16349138 TI - Mobile bacteria and transport of polynuclear aromatic hydrocarbons in porous media. AB - [This corrects the article on p. 3311 in vol. 59.]. PMID- 16349140 TI - Direct Polymerase Chain Reaction Detection of Campylobacter jejuni and Campylobacter coli in Raw Milk and Dairy Products. AB - [This corrects the article on p. 2161 in vol. 59.]. PMID- 16349141 TI - Degradation and Induction Specificity in Actinomycetes That Degrade p Nitrophenol. AB - [This corrects the article on p. 3506 in vol. 59.]. PMID- 16349142 TI - New Medium for the Simultaneous Detection of Total Coliforms and Escherichia coli in Water. AB - [This corrects the article on p. 3534 in vol. 59.]. PMID- 16349143 TI - Utilization of Lactic Acid by Fusarium oxysporum var. lini: Regulation of Transport and Metabolism. AB - Lactic acid was transported in Fusarium oxysporum var. lini ATCC 10960 by a saturable transport system that had a half-saturation constant of 56.6 +/- 7.5 muM and a maximum velocity of 0.61 +/- 0.10 mmol h g (dry weight) at 26 degrees C and pH 5.0. This transport system was inducible and was not expressed in the presence of a repressing substrate. Evidence is presented that the anionic form lactate was taken up by the cells. Propionic, acetic, pyruvic, and bromoacetic acids but not succinic acid competitively inhibited the transport of lactic acid. Bromoacetic acid, which was not metabolized, was taken up to a steady-state level when intracellular and extracellular concentrations were identical, indicating that the transport system was not accumulative. The enzymatic activity that was physiologically more relevant in the metabolism of lactic acid was lactate: ferricytochrome c oxidase. This enzyme did not exhibit stereospecifity and was induced by lactic acid. PMID- 16349144 TI - Inhibition of aflatoxin production by surfactants. AB - The effect of 12 surfactants on aflatoxin production, growth, and conidial germination by the fungus Aspergillus flavus is reported. Five nonionic surfactants, Triton X-100, Tergitol NP-7, Tergitol NP-10, polyoxyethylene (POE) 10 lauryl ether, and Latron AG-98, reduced aflatoxin production by 96 to 99% at 1% (wt/vol). Colony growth was restricted by the five nonionic surfactants at this concentration. Aflatoxin production was inhibited 31 to 53% by lower concentrations of Triton X-100 (0.001 to 0.0001%) at which colony growth was not affected. Triton X-301, a POE-derived anionic surfactant, had an effect on colony growth and aflatoxin production similar to that of the five POE-derived nonionic surfactants. Sodium dodecyl sulfate (SDS), an anionic surfactant, and dodecyltrimethylammonium bromide, a cationic surfactant, suppressed conidial germination at 1% (wt/vol). SDS had no effect on aflatoxin production or colony growth at 0.001%. The degree of aflatoxin inhibition by a surfactant appears to be a function of the length of the hydrophobic and hydrophilic chains of POE derived surfactants. PMID- 16349145 TI - Phase Variation in Xenorhabdus nematophilus and Photorhabdus luminescens: Differences in Respiratory Activity and Membrane Energization. AB - Phase variation in Xenorhabdus and Photorhabdus spp. has a significant impact on their symbiotic relationship with entomopathogenic nematodes by altering the metabolic by-products upon which the nematodes feed. The preferential retention of the phase I variant by the infective-stage nematode and its better support for nematode reproduction than phase II indicates its importance in the bacterial nematode interactions. However, there is no obvious role for phase II in these interactions. This study has revealed differences in the respiratory activity between the two phases of Xenorhabdus nematophilus A24 and Photorhabdus luminescens Hm. After experiencing periods of starvation, phase II cells recommenced growth within 2 to 4 h from the addition of nutrients, compared with 14 h for phase I cells, indicating a more efficient nutrient uptake ability in the former. The levels of activity of major respiratory enzymes were 15 to 100% higher in phase II cells from stationary cultures in complex media than in phase I cells. Transmembrane proton motive force measurements were also higher by 20% in phase II under the same conditions. The increased membrane potentials reflect upon the ability of the phase II variant to respond to nutrients, both through growth and nutrient uptake. It is postulated that while phase I cells are better adapted to conditions in the insect and the nematode, phase II cells may be better adapted to conditions in soil as free-living organisms. PMID- 16349146 TI - Stable Expression of hom-1-thrB in Corynebacterium glutamicum and Its Effect on the Carbon Flux to Threonine and Related Amino Acids. AB - The hom-1-thrB operon encodes homoserine dehydrogenase resistant to feedback inhibition by L-threonine and homoserine kinase. Stable expression of this operon has not yet been attained in different Corynebacterium glutamicum strains. We studied the use of chromosomal integration and of a low-copy-number vector for moderate expression of the hom-1-thrB operon to enable an analysis of the physiological consequences of its expression in C. glutamicum. Strains carrying one, two, or three copies of hom-1-thrB were obtained. They showed proportionally increased enzyme activity of feedback-resistant homoserine dehydrogenase and of homoserine kinase. This phenotype was stably maintained in all recombinants for more than 70 generations. In a lysine-producing C. glutamicum strain which does not produce any threonine, expression of one copy of hom-1-thrB resulted in the secretion of 39 mM threonine. Additional copies resulted in a higher, although not proportional, accumulation of threonine (up to 69 mM). This indicates further limitations of threonine production. As the copy number of hom-1-thrB increased, increasing amounts of homoserine (up to 23 mM) and isoleucine (up to 34 mM) were secreted. Determination of the cytosolic concentration of the respective amino acids revealed an increase of intracellular threonine from 9 to 100 mM and of intracellular homoserine from 4 to 74 mM as the copy number of hom-1-thrB increased. These results suggest that threonine production with C. glutamicum is limited by the efflux system for this amino acid. Furthermore, the results show the successful use of moderate and stable hom-1-thrB expression for directing the carbon flux from aspartate to threonine. PMID- 16349147 TI - Purification and Characterization of an l-Amino Amidase from Mycobacterium neoaurum ATCC 25795. AB - An l-amino amidase from Mycobacterium neoaurum ATCC 25795 responsible for the enantioselective resolution of dl-alpha-methyl valine amide was purified and characterized. The purification procedure included ammonium sulfate fractionation, gel filtration, and anion-exchange chromatography, which resulted in a homogeneous preparation of the enzyme with a native molecular mass of 136 kDa and a subunit molecular mass of 40 kDa. The purified enzyme displayed the highest activity at 50 degrees C and at pH 8.0 and 9.5. The enzyme was strongly inhibited by the metal-chelating agent 1,10-phenanthroline, the disulfide reducing agent dithiothreitol, and the cysteine proteinase inhibitor iodoacetamide. The purified amino amidase showed a unique l-enantioselective activity towards a broad range of both alpha-H- and alpha-alkyl-substituted amino acid amides, with the highest activity towards the cyclic amino acid amide dl proline amide. No activity was measured with dl-mandelic acid amide nor with the dipeptide l-phenylalanine-l-leucine. The highest catalytic efficiency (k(cat)/K(m) ratio) was measured with dl-alpha-allyl alanine amide, dl-alpha methyl phenylalanine amide, and dl-alpha-methyl leucine amide. PMID- 16349148 TI - Quantitative centrifugation to extract benthic protozoa from freshwater sediments. AB - TWO METHODS FOR EXTRACTING PROTISTS FROM FRESHWATER SEDIMENT ARE DESCRIBED: (i) an adapted isopycnic centrifugation technique for sandy and gyttja-like sediments and (ii) a rate zonal centrifugation technique for sediments rich in particulate organic material (litter-like sediments). The recoveries of protists during isopycnic centrifugation in media of several densities were compared. No significant losses in sodium diatrizoate and Percoll were recorded. After known amounts of nanoflagellates were added to azoic sediments, the protists were extracted and counted. For sandy sediments, we found 100% recovery, and for the gyttja-like sediments we found a maximum recovery of 94%. The recovery of protozoa extracted from litter-like sediments, characteristic of littoral systems, depends on a given centrifugal force, on time, and on the dimensions of the flagellates. A recovery model which takes into account cell dimensions and centrifugation characteristics gives the minimum expected recovery. PMID- 16349149 TI - Isolation of Bacillus thuringiensis from Stored Tobacco and Lasioderma serricorne (F.). AB - Bacillus thuringiensis was isolated from dried tobacco residues and dead tobacco beetles (Lasioderma serricorne (F.); Coleoptera: Anobiidae) collected in a large number of locations worldwide. Eighty-eight samples of stored tobacco were analyzed and yielded 78 B. thuringiensis strains which were characterized on the basis of parasporal crystal morphology, sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles, and the results of an immunoblot analysis of the insecticidal crystal proteins. Flagellar antigen identification was used to differentiate selected isolates. Strains that produced rhomboidal crystals associated with the Coleoptera-specific pathotype (Cry III group) were the most abundant strains (59% of the isolates). Preliminary toxicity assays were performed with L. serricorne larvae, and the results suggested that activity is not restricted to isolates related to the Coleoptera-specific group. The results of our survey indicate that B. thuringiensis is part of the natural microflora in the stored-tobacco environment and that this special habitat represents a source of B. thuringiensis isolates that may be used to control stored-product pests. PMID- 16349150 TI - Evaluation of data transformations and validation of a model for the effect of temperature on bacterial growth. AB - The temperature of chilled foods is an important variable for controlling microbial growth in a production and distribution chain. Therefore, it is essential to model growth as a function of temperature in order to predict the number of organisms as a function of temperature and time. This article deals with the correct variance-stabilizing transformation of the growth parameters A (asymptotic level), mu (specific growth rate), and lambda (lag time). This is of importance for the regression analysis of the data. A previously gathered data set and model for the effect of temperature on the growth of Lactobacillus plantarum (M. H. Zwietering, J. T. de Koos, B. E. Hasenack, J. C. de Wit, and K. van 't Riet, Appl. Environ. Microbiol. 57:1094-1101, 1991) is extended with new data. With the total data set (original and new data), a variance-stabilizing transformation is selected in order to determine which transformation should precede fitting. No transformation for the asymptote data, a square root for the growth rate, and a logarithmic transformation for the lag time were found to be appropriate. After these transformations, no significant correlation was found between the variance and the magnitude of the variable. Model corrections were made and model parameters were estimated by using the original data. With the new data, the models were validated by comparing the lack of fit of the models with the measurement error, using an F test. The predictions of the models for mu and lambda were adequate. The model for A showed a systematic deviation, and therefore a new model for A is proposed. PMID- 16349151 TI - Modeling of bacterial growth with shifts in temperature. AB - The temperature of chilled foods is an important variable for the shelf life of a product in a production and distribution chain. To predict the number of organisms as a function of temperature and time, it is essential to model the growth as a function of temperature. The temperature is often not constant in various stages of distribution. The objective of this research was to determine the effect of shifts in temperature. The suitability and usefulness of several models to describe the growth of Lactobacillus plantarum with fluctuating temperatures was evaluated. It can be assumed that temperature shifts within the lag phase can be handled by adding relative parts of the lag time to be completed and that temperature shifts within the exponential phase result in no lag phase. With these assumptions, the kinetic behavior of temperature shift experiments was reasonably well predicted, and this hypothesis was accepted statistically in 73% of the cases. Only shifts of temperature around the minimum temperature for growth showed very large deviations from the model prediction. The best results were obtained with the assumption that a temperature shift (within the lag phase as well as within the exponential phase) results in an additional lag phase. This hypothesis was accepted statistically in 93% of the cases. The length of the additional lag phase is one-fourth of the lag time normally found at the temperature after the shift. PMID- 16349153 TI - Selective Desulfurization of Dibenzothiophene by Rhodococcus erythropolis D-1. AB - A dibenzothiophene (DBT)-degrading bacterium, Rhodococcus erythropolis D-1, which utilized DBT as a sole source of sulfur, was isolated from soil. DBT was metabolized to 2-hydroxybiphenyl (2-HBP) by the strain, and 2-HBP was almost stoichiometrically accumulated as the dead-end metabolite of DBT degradation. DBT degradation by this strain was shown to proceed as DBT --> DBT sulfone --> 2-HBP. DBT at an initial concentration of 0.125 mM was completely degraded within 2 days of cultivation. DBT at up to 2.2 mM was rapidly degraded by resting cells within only 150 min. It was thought this strain had a higher DBT-desulfurizing ability than other microorganisms reported previously. PMID- 16349152 TI - Introduction of a Lepidopteran-Specific Insecticidal Crystal Protein Gene of Bacillus thuringiensis subsp. kurstaki by Conjugal Transfer into a Bacillus megaterium Strain That Persists in the Cotton Phyllosphere. AB - A lepidopteran toxin gene of the entomopathogen Bacillus thuringiensis subsp. kurstaki HD-1 was introduced into a cotton leaf-colonizing Bacillus megaterium strain, RS1, by conjugal transfer. Rifampin- and nalidixic acid-resistant colonies obtained after cell mating were screened for crystal production by microscopy. A transcipient, B. megaterium RS1-43, was selected by this procedure. Southern blot hybridization with both total DNA and HindIII-digested DNA of the transcipient showed positive signals with a cryIA-specific probe, suggesting the transfer of the lepidopteran-specific cryIA(a) gene. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblot analysis confirmed the presence of the 134-kDa toxic crystal protein specific to lepidopteran larvae in the transcipient. Survival studies with cultures of the transcipient at both vegetative and postvegetative growth stages on cotton, under field conditions, suggested that the bacterium persisted on the leaf surfaces for more than 28 days, with a gradual decline in the population level with time, while the donor, B. thuringiensis subsp. kurstaki, disappeared completely after 7 days following inoculation. An in situ differential crystal-staining technique indicated the production of crystals by the transcipient on cotton leaf surfaces for about 30 days. Leaf bioassays of cotton plants inoculated with a single spray of the transcipient showed 75- to 96% mortality to the first-instar larvae of Heliothis armigera up to 21 days, and this single spray conferred total protection to the plants for about 30 days by causing an antifeeding effect on the remaining larvae. PMID- 16349154 TI - Characterization of microbial community structure in the surface sediment of osaka bay, Japan, by phospholipid Fatty Acid analysis. AB - Twenty-eight sediment samples collected from Osaka Bay, Japan, were analyzed for phospholipid ester-linked fatty acids (PLFA) to determine regional differences in microbial community structure of the bay. The abundance of three major groups of C(10) to C(19) PLFA (saturated, branched, and monounsaturated PLFA), which accounted for 84 to 97% of the total PLFA, indicated the predominance of prokaryotes in the sediment. The distribution of six clusters obtained by similarity analysis in the bay revealed a marked regional distribution in the PLFA profiles. Total PLFA concentrations (0.56 to 2.97 mug/g [dry weight] of the sediment) in sediments also showed marked variation among the stations, with higher concentrations of total PLFA in the central part of the bay. The biomass, calculated on the basis of total PLFA concentration, ranged from 0.25 x 10 to 1.35 x 10 cells per g (dry weight) of the sediment. The relative dominance of microbial groups in sediments was described by using the reported bacterial biomarker fatty acids. Very small amounts of the characteristic PLFA of microeukaryotes in sediments indicated the restricted distribution of microeukaryotes. By examining the distribution of clusters and groups of microorganisms in the bay, there were two characteristics of the distribution pattern: (i) the predominance of anaerobic bacteria and gram-positive prokaryotes, characterized by the high proportions of branched PLFA in the eastern and northeastern sides of the bay, where the reported concentrations of pollutants were also high, and (ii) the predominance of aerobic prokaryotes and eukaryotes, except for a few stations, in the western and southwestern sides of the bay, as evidenced by the large amounts of monounsaturated PLFA. Such significant regional differences in microbial community structure of the bay indicate shifts in microbial community structure. PMID- 16349155 TI - Electrophoretic transfer from polyacrylamide gel to nitrocellulose sheets, a new method to characterize multilocus enzyme genotypes of Klebsiella strains. AB - A new method for multilocus enzyme electrophoresis, based on electrophoretic transfers to nitrocellulose after polyacrylamide-agarose gel electrophoresis was explored. Electrophoretic separation was performed on 1-mm-thick slab gels with 6 mul samples of bacterial extracts and was followed by serial 5-min consecutive transfers. The transferability of 19 metabolic enzymes of Klebsiella strains was studied and allowed the simultaneous examination of one enzyme in the separation gel and at least five enzymes on nitrocellulose sheets. The resolution of enzyme bands was increased on nitrocellulose; thus, well-separated bands were recorded for nucleoside phosphorylase, peptidase, and phosphoglucose isomerase whereas their mobility variants could not be clearly distinguished in the separation gel because of stain diffusion. The study of genetic relationships of 42 strains of Klebsiella pneumoniae and 24 strains of Klebsiella oxytoca demonstrated the reliability of the method, since clustering analysis of electrophoretic types, based on electrophoretic polymorphism of 10 metabolic enzymes, showed two main clusters well correlated with the two species. The 57 electrophoretic types described confirm the usefulness of the method for the study of genetic relationships between closely related strains. PMID- 16349156 TI - Significant biogenesis of chlorinated aromatics by fungi in natural environments. AB - Common wood- and forest litter-degrading fungi produce chlorinated anisyl metabolites. These compounds, which are structurally related to xenobiotic chloroaromatics, occur at high concentrations of approximately 75 mg of chlorinated anisyl metabolites kg of wood or litter in the environment. The widespread ability among common fungi to produce large amounts of chlorinated aromatic compounds in the environment makes us conclude that these kinds of compounds can no longer be considered to originate mainly from anthropogenic sources. PMID- 16349157 TI - Physiological Role of Chlorinated Aryl Alcohols Biosynthesized De Novo by the White Rot Fungus Bjerkandera sp. Strain BOS55. AB - The white rot fungus Bjerkandera sp. strain BOS55 produces veratryl, anisyl, 3 chloroanisyl, and 3,5-dichloroanisyl alcohol and the corresponding aldehydes de novo from glucose. All metabolites are produced simultaneously with the extracellular ligninolytic enzymes and have an important physiological function in the fungal ligninolytic system. Both mono- and dichlorinated anisyl alcohols are distinctly better substrates for the extracellular aryl alcohol oxidases than veratryl alcohol. The aldehydes formed are readily recycled by reduction by washed fungal mycelium, thus creating an extracellular H(2)O(2) production system regulated by intracellular enzymes. Lignin peroxidase does not oxidize the chlorinated anisyl alcohols either in the absence or in the presence of veratryl alcohol. It was therefore concluded that the chlorinated anisyl alcohols are well protected against the fungus's own aggressive ligninolytic enzymes. The relative amounts of veratryl alcohol and the chlorinated anisyl alcohols differ significantly according to the growth conditions, indicating that production of veratryl alcohol and the production of the (chlorinated) anisyl metabolites are independently regulated. We conclude that the chlorinated anisyl metabolites biosynthesized by the white rot fungus Bjerkandera sp. strain BOS55 can be purposefully produced for ecologically significant processes such as lignin degradation. PMID- 16349158 TI - A gas lift bioreactor for removal of contaminants from the vapor phase. AB - The cometabolic degradation of trichloroethylene (TCE) as a vapor by two aromatic metabolizing pseudomonads was evaluated in an airlift reactor. These microorganisms were able to degrade 90 to 95% of TCE in air at concentrations at the reactor inlet of 300 to 4,000 mug/liter. Although exposure of the cells to high inlet concentrations of TCE (4 mg/liter) caused a decline in enzyme-specific activity and TCE removal efficiency, this loss in activity could be prevented or delayed by increasing the rate of cosubstrate addition. Under the appropriate operating conditions, the microorganisms were able to degrade even high concentrations of TCE and activity of the cells in the reactor could be maintained for periods of at least 2 weeks. PMID- 16349159 TI - Nitrate Reduction in a Sulfate-Reducing Bacterium, Desulfovibrio desulfuricans, Isolated from Rice Paddy Soil: Sulfide Inhibition, Kinetics, and Regulation. AB - From the second-highest dilution in a most-probable-number dilution series with lactate and sulfate as substrates and rice paddy soil as the inoculum, a strain of Desulfovibrio desulfuricans was isolated. In addition to reducing sulfate, sulfite, and thiosulfate, the strain also reduced nitrate to ammonia. The latter process was studied in detail, since the ability to reduce nitrate was strongly influenced by the presence of sulfide. Sulfide inhibited both growth on nitrate and nitrate reduction. A 70% inhibition of the nitrate reduction rate was obtained at 127 muM sulfide, and growth was inhibited by 50% at approximately 320 muM sulfide and was not detectable above 700 muM sulfide. In contrast, sulfate reduction was not affected at concentrations of up to 5 mM. After growth with sulfate, an induction period of 2 to 4 days was needed before nitrate reduction started. When nitrate and sulfate were present simultaneously, only sulfate was reduced, except when sulfate was present at very low concentrations (4 muM). At higher sulfate concentrations (500 muM), nitrate reduction was temporarily halted. The affinity for nitrate uptake was extremely high (K(m) = 0.05 muM) compared with that for sulfate uptake (K(m) = 5 muM). Thus, at low nitrate concentrations this bacterium is favored relative to denitrifiers (K(m) = 1.8 to 13.7 muM) or other nitrate ammonifiers (e.g., Clostridium spp. [K(m) = 500 muM]). PMID- 16349160 TI - Changes in Bacterial Numbers and Leucine Assimilation during Estimations of Microbial Respiratory Rates in Seawater by the Precision Winkler Method. AB - During incubation of seawater in bottles, the decrease in dissolved oxygen is often nonlinear over time scales frequently used to measure respiration. Numbers of bacteria always increase, and rates of assimilation of dissolved leucine often increase exponentially. This suggests that sample handling disrupts the previously existing food web, leading to shifts of trophic state and unbalanced growth. Potential errors in measuring respiratory rate can be minimized by documenting these variables. PMID- 16349161 TI - Inactivation of Phage MS2 by Iron-Aided Titanium Dioxide Photocatalysis. AB - Near-UV photocatalytic disinfection was accomplished in aqueous titanium dioxide suspensions. A level of inactivation of phage MS2 of 90% increased to 99.9% after 2 muM ferrous sulfate was added. Hydroxyl radical oxidation, with Fenton reaction enhancement, is suspected to be primarily responsible for the viral degradation observed. PMID- 16349162 TI - Pseudomonas putida Strains Which Constitutively Overexpress Mercury Resistance for Biodetoxification of Organomercurial Pollutants. AB - Improved biocatalysts for mercury (Hg) remediation were generated by random mutagenesis of Pseudomonas putida with a minitransposon containing merTPAB, the structural genes specifying organomercury resistance. Subsequent selection for derivatives exhibiting elevated resistance levels to phenylmercury allowed the isolation of strains that constitutively express merTPAB at high levels, conferring the ability to cleave Hg from an organic moiety and reduce the freed Hg(II) to the less toxic elemental form, Hg, at greater rates. Constitutive overexpression of merTPAB had no apparent effect on culture growth rates, even when Hg(II) was initially present at otherwise toxic concentrations. These properties were also combined with benzene and toluene catabolism, allowing detoxification of the metal component of phenyl mercuric acetate, as well as degradation of its aromatic moiety. PMID- 16349163 TI - Microbial degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3 hydroxyvalerate) in soils. AB - [This corrects the article on p. 3236 in vol. 59.]. PMID- 16349164 TI - Intracellular Concentrations of Coenzyme A and Its Derivatives from Clostridium acetobutylicum ATCC 824 and Their Roles in Enzyme Regulation. AB - Intracellular levels of coenzyme A (CoA) and its derivatives involved in the metabolic pathways for Clostridium acetobutylicum ATCC 824 were analyzed by using reverse-phase high-performance liquid chromatography (HPLC). During the shift from the acidogenic to the solventogenic or stationary growth phase, the concentration of butyryl-CoA increased rapidly and the concentrations of free CoA and acetyl-CoA decreased. These changes were accompanied by a rapid increase of the solvent pathway enzyme activity and a decrease of the acid pathway enzyme activity. Assays with several non-solvent-producing mutant strains were also carried out. Upon entry of the mutant strains to the stationary phase, the butyryl-CoA concentrations for these mutant strains were comparable to those for the wild type even though the mutants were deficient in solvent-producing enzymes. Levels of acetoacetyl-CoA, beta-hydroxy-butyryl-CoA, and crotonyl-CoA compounds in both wild-type and mutant extracts were below HPLC detection thresholds (<21 muM). PMID- 16349165 TI - Rapid Identification of Rhizobia by Restriction Fragment Length Polymorphism Analysis of PCR-Amplified 16S rRNA Genes. AB - Forty-eight strains representing the eight recognized Rhizobium species, two new Phaseolus bean Rhizobium genomic species, Bradyrhizobium spp., Agrobacterium spp., and unclassified rhizobia from various host plants were examined by restriction fragment length polymorphism (RFLP) analysis of 16S rRNA genes amplified by polymerase chain reaction (PCR). Twenty-one composite genotypes were obtained from the combined data of the RFLP analysis with nine endonucleases. Species assignments were in full agreement with the established taxonomic classification. Estimation from these data of genetic relationships between and within genera and species correlated well with previously published data based on DNA-rRNA hybridizations and sequence analysis of 16S rRNA genes. This PCR-RFLP method provides a rapid tool for the identification of root nodule isolates and the detection of new taxa. PMID- 16349166 TI - Identification of a Genetic Locus in Pseudomonas aureofaciens Involved in Fungal Inhibition. AB - In iron-rich conditions, Pseudomonas aureofaciens PA147-2 produces an antibiotic like compound that inhibits the growth of a plant fungal pathogen, Aphanomyces euteiches. To contribute to the potential use of PA147-2 as a biocontrol organism, we report the identification of a genetic locus important for antibiotic biosynthesis. Mutants defective for fungal inhibition (Af) were generated by Tn5 mutagenesis. Southern hybridization of total DNAs from three Af mutants indicated that loss of fungal inhibition was due to a single Tn5 insertion in each mutant. Restriction mapping of the mutation points showed that in two mutants the Tn5 insertions were in the same 16.0-kb EcoRI fragment and were separated by 2.1 kb. A genomic library of PA147-2 was constructed and screened by using a region of DNA flanking the Tn5 insertion in one mutant (PA109) as a probe to recover complementing cosmids. Three cosmids containing a 16.0-kb EcoRI fragment complementary to the two mutants were recovered. Allele replacement by homologous recombination with putative complementing cosmids restored one mutant to antifungal activity against A. euteiches. Southern analysis of the complemented mutants confirmed that allele replacement had occurred between cosmid DNA and Tn5. The wild-type 16.0-kb EcoRI fragment was cloned from the cosmid and complemented the two mutants to antifungal activity. An antifungal compound was isolated from PA147-2 grown on solid medium. Antifungal activity correlated to a peak on high-pressure liquid chromatography analysis. Under the same growth and extraction conditions, the antifungal activity seen in PA147-2 was absent in two Af mutants. Furthermore, absence of an antifungal compound in each mutant correlated to the absence of the wild-type "antifungal" peak on high-pressure liquid chromatography analysis. PMID- 16349167 TI - Cloning of Genes Involved in the Synthesis of Pyrrolnitrin from Pseudomonas fluorescens and Role of Pyrrolnitrin Synthesis in Biological Control of Plant Disease. AB - A soil isolate of Pseudomonas fluorescens (BL915) was shown to be an effective antagonist of Rhizoctonia solani-induced damping-off of cotton. Investigation of the biological basis of this antagonism revealed that the strain produces pyrrolnitrin, a secondary metabolite known to inhibit R. solani and other fungi. Mutants of strain BL915 that did not produce pyrrolnitrin and did not suppress damping-off of cotton by R. solani were generated by exposure to N-methyl-N' nitro-N-nitrosoguanidine. A gene region that was capable of restoring pyrrolnitrin production to the non-pyrrolnitrin-producing mutants and of conferring this ability upon two other P. fluorescens strains not otherwise known to produce this compound or to be capable of suppressing damping-off caused by R. solani was isolated from strain BL915. The non-pyrrolnitrin-producing strains (mutants of BL915 and the other two P. fluorescens strains) which synthesized pyrrolnitrin after the introduction of the gene region from strain BL915 were also shown to be equal to strain BL915 in their ability to suppress R. solani induced damping-off of cotton. These results indicate that we have isolated from P. fluorescens BL915 a gene(s) that has a role in the synthesis of pyrrolnitrin and that the production of this compound has a role in the ability of this strain to control damping-off of cotton by R. solani. PMID- 16349168 TI - Effects of DNA polymer length on its adsorption to soils. AB - Three different DNA fragments ranging size from 2.69 kbp (1.75 MDa) to 23 kbp (14.95 MDa) were used as tracers to study the adsorption of polydisperse solutions of calf thymus DNA to eight model soils. The adsorption of the three tracers to all soils was described by the Freundlich adsorption model, with adsorption coefficients (K) ranging from 1.1 for acid-washed sand to over 300 for one soil. An inverse relationship between tracer size and K was observed with six of the eight soils, indicating that smaller fragments are sorbed preferentially versus larger fragments in these soils. No significant correlation between K and the organic carbon contents, clay contents, pHs, or cation exchange capacities of the model soils was observed. PMID- 16349169 TI - Tyrosinase reaction and subsequent chitosan adsorption for selective removal of a contaminant from a fermentation recycle stream. AB - In the industrial production of penicillin V, the phenoxyacetate precursor is added to the fermentor to direct biosynthesis. When used for producing semisynthetic penicillins, the penicillin V is often hydrolyzed to 6 aminopenicillanic acid with the regeneration of the phenoxyacetate precursor. To reduce raw-material as well as waste-disposal costs, it is desirable to recycle the phenoxyacetate precursor. Unfortunately, the recycle stream is generally contaminated by the p-hydroxylated derivative of this precursor. We examined a two-step approach to eliminate this contaminant. In the first step the tyrosinase enzyme was used to selectively convert the p-hydroxyphenoxyacetate contaminant to a reactive intermediate-presumably its quinone. In the second step, the tyrosinase-generated reactive intermediate was allowed to react with and strongly bind to chitosan. In contrast, the phenoxyacetate precursor was neither oxidized by tyrosinase nor bound to chitosan. When concentrated phenoxyacetate solutions were tested, the combination of tyrosinase and chitosan effectively converted low levels of the p-hydroxyphenoxyacetate contaminant and removed its products from solution, while the concentration of the phenoxyacetate precursor was unaffected. PMID- 16349170 TI - Serological and Ecological Characteristics of a Nodule-Dominant Serotype from an Indigenous Soil Population of Rhizobium leguminosarum bv. trifolii. AB - Although at least 13 antigenically distinct serotypes of Rhizobium leguminosarum bv. trifolii exist in an Abiqua silty clay loam soil, one serotype, AS6, occupies >/=50% of the root nodules formed on field-grown subclover and between 33 and 78% of the nodules formed on five annual clover species grown in the same soil under laboratory conditions. The dominance of subclover nodules by serotype AS6 was reproducible over a 4-year sampling period and throughout the entire 200- by 100 m pasture examined. Serotype AS6 was composed of three antigenically distinct subtypes (AS6-a, AS6-b, and AS6-c). Each subtype contributed about one-third of the AS6 isolates recovered from nodules of field-grown subclover plants and maintained similar population densities in nonrhizosphere and rhizosphere soil. Rhizobia with the AS6 antigenic signature accounted for from 20 to 100% of the soil populations of R. leguminosarum in arable and pasture soils under legumes throughout the state of Oregon. Over a 12-month period, the population densities of the serotype AS6 complex and three minor nodule-occupying serotypes (AG4, AP17, and AS21) were measured in the rhizospheres of field-grown subclover and orchard grass and in nonrhizosphere Abiqua soil. Regardless of season or serotype, the orchard grass rhizosphere effect was minimal, with the ratio between rhizosphere and nonrhizosphere serotype population densities ranging between 2.5 (midsummer) and 10.5 (spring). In contrast, the magnitude of the subclover rhizosphere effect varied seasonally and among serotypes. Between October and December the ratios for all serotypes were similar (12.5 to 25.5). However, in the spring (April and May), the magnitude of the rhizosphere effect varied among the indigenous serotypes (ratios, 10.5 to 442) and for minor nodule occupying serotypes AS21 (ratio, 442) and AP17 (ratio, 47) was as great as, or even greater than, the magnitude of the rhizosphere effect observed with the AS6 complex (ratio, 65.5). PMID- 16349171 TI - Genotypic and Phenotypic Comparisons of Chromosomal Types within an Indigenous Soil Population of Rhizobium leguminosarum bv. trifolii. AB - The relative genetic similarities of 200 isolates of Rhizobium leguminosarum bv. trifolii recovered from an Oregon soil were determined at 13 enzyme loci by multilocus enzyme electrophoresis (MLEE). These isolates represented 13 antigenically distinct serotypes recovered from nodules formed on various clover species. The MLEE-derived levels of relatedness among isolates of R. leguminosarum bv. trifolii were found to be in good agreement with the levels of relatedness established by using repetitive (repetitive extragenic palindromic and enterobacterial repetitive intergeneric consensus) sequences and the PCR technique and with levels of relatedness from previously published DNA reassociation studies. BIOLOG substrate utilization patterns showed that isolates within an electrophoretic type (ET) were phenotypically more similar to each other than to isolates of other ETs. The soil isolates were represented by 53 ETs which could be clustered into seven groups (groups B, E, G, H1, H2, I, and J). Evidence for multilocus structure within the population was obtained, and group B was identified as the primary creator of the disequilibrium. Of 75 isolates belonging to the nodule-dominant serotype AS6 complex, 72 were found in group B. Isolates WS2-01 and WS2-02 representing nodule-dominant serotypes recovered from subclover grown at another Oregon site were also found in group B. Isolates representing the most numerous ETs in group B (ETs 2 and 3) were either suboptimally effective or completely ineffective at fixing nitrogen on six different clover species. Another four groups of isolates (groups A, C, D, and F) were identified when 32 strains of diverse origins were analyzed by MLEE and incorporated into the cluster analysis. Group A was most dissimilar in comparisons with other groups and contained strain USDA 2124 (T24), which produces trifolitoxin and has unique symbiotic characteristics. PMID- 16349172 TI - Symbiotic Characteristics of Rhizobium leguminosarum bv. trifolii Isolates Which Represent Major and Minor Nodule-Occupying Chromosomal Types of Field-Grown Subclover (Trifolium subterraneum L.). AB - The symbiotic effectiveness and nodulation competitiveness of Rhizobium leguminosarum bv. trifolii soil isolates were evaluated under nonsoil greenhouse conditions. The isolates which we used represented both major and minor nodule occupying chromosomal types (electrophoretic types [ETs]) recovered from field grown subclover (Trifolium subterraneum L.). Isolates representing four ETs (ETs 2, 3, 7, and 8) that were highly successful field nodule occupants fixed between 2- and 10-fold less nitrogen and produced lower herbage dry weights and first harvest herbage protein concentrations than isolates that were minor nodule occupants of field-grown plants. Despite their equivalent levels of abundance in nodules on field-grown subclover plants, ET 2 and 3 isolates exhibited different competitive nodulation potentials under nonsoil greenhouse conditions. ET 3 isolates generally occupied more subclover nodules than isolates belonging to other ETs when the isolates were mixed in 1:1 inoculant ratios and inoculated onto seedlings. In contrast, ET 2 isolates were less successful at nodulating under these conditions. In many cases, ET 2 isolates required a numerical advantage of at least 6:1 to 11:1 to occupy significantly more nodules than their competitors. We identified highly effective isolates that were as competitive as the ET 3 isolates despite representing serotypes that were rarely recovered from nodules of field-grown plants. When one of the suboptimally effective isolates (ET2-1) competed with an effective and competitive isolate (ET31-5) at several different inoculant ratios, the percentages of nodules occupied by the former increased as its numerical advantage increased. Although subclover yields declined as nodule occupancy by ET2-1 increased, surprisingly, this occurred at inoculant ratios at which large percentages of nodules were still occupied by ET31-5. PMID- 16349173 TI - Multicellular organization in a degradative biofilm community. AB - Diclofop methyl, a commercial herbicide, was used as the sole carbon source to cultivate diclofop-degrading biofilms in continuous-flow slide culture. The biofilms were analyzed by using scanning confocal laser microscopy and image analysis. Spatial relationships among members of the community were distinctive to diclofop-grown biofilms. These relationships did not develop when the biofilms were grown on more labile substrates but were conserved when the biofilms were cultivated with other chlorinated ring compounds. The structures included conical bacterial consortia rising to 30 mum above the surrounding biofilm, grape-like clusters of cocci embedded in a matrix of perpendicularly oriented bacilli, and other highly specific patterns of intra- and intergeneric cellular coaggregation and growth. These unique consortial relationships indicated that syntrophic interactions may be necessary for optimal degradation of diclofop methyl and other chlorinated ring compounds. PMID- 16349174 TI - Nucleation of celestite and strontianite on a cyanobacterial s-layer. AB - Synechococcus strain GL24 is a unicellular cyanobacterium that was isolated from Fayetteville Green Lake, New York, a meromictic lake which has high Ca and SO(4) concentrations. Epicellular mineralization of Synechococcus cells in the lake is the mechanism by which extensive calcitic bioherms (or microbial reefs) have been formed on the lake's shore and a marl sediment has been built on the lake bottom. Previous studies have shown that calcium carbonate (calcite) formation on the Synechococcus surface is dependent upon an alkaline pH, which is produced in the cellular microenvironment by the cells as their activity increases with seasonal warming of the lake water. At the circumneutral pH of bulk lake water, calcium sulfate (gypsum) is formed. In this study, we show that Synechococcus mediates a similar sulfate-to-carbonate transformation when Sr is the major divalent cation present, forming celestite and strontianite. In experimental systems to which equimolar amounts of Ca and Sr, Ca or Sr and Mg, or all three ions together were added to artificial lake water, Ca and Sr were incorporated equally into mineral formation to form CaSr(CO(3))(2). No Mg -containing carbonates were formed when either or both of the other two ions were present. Mineral formation takes place on a hexagonally arranged proteinaceous template (an S-layer) which forms the outermost surface of the Synechococcus cell. Our results provide evidence that the S-layer exhibits selectivity with respect to the ions bound and subsequently incorporated into carbonate minerals and that celestite and strontianite, previously thought to be purely evaporitic minerals, can be biogenically formed. PMID- 16349175 TI - Ecology of Thermophilic Fungi in Mushroom Compost, with Emphasis on Scytalidium thermophilum and Growth Stimulation of Agaricus bisporus Mycelium. AB - Twenty-two species of thermophilic fungi were isolated from mushroom compost. Scytalidium thermophilum was present in the compost ingredients, fresh straw, horse droppings, and drainage from compost and dominated the fungal biota of compost after preparation. Of 34 species of thermophilic fungi tested, 9 promoted mycelial growth of Agaricus bisporus on sterilized compost: Chaetomium thermophilum, an unidentified Chaetomium sp., Malbranchea sulfurea, Myriococcum thermophilum, S. thermophilum, Stilbella thermophila, Thielavia terrestris, and two unidentified basidiomycetes. These species will be considered for future experiments on inoculation and more controlled preparation of compost. PMID- 16349176 TI - Lignin peroxidase oxidation of aromatic compounds in systems containing organic solvents. AB - Lignin peroxidase from Phanerochaete chrysosporium was used to study the oxidation of aromatic compounds, including polycyclic aromatic hydrocarbons and heterocyclic compounds, that are models of moieties of asphaltene molecules. The oxidations were done in systems containing water-miscible organic solvents, including methanol, isopropanol, N, N-dimethylformamide, acetonitrile, and tetrahydrofuran. Of the 20 aromatic compounds tested, 9 were oxidized by lignin peroxidase in the presence of hydrogen peroxide. These included anthracene, 1-, 2 , and 9-methylanthracenes, acenaphthene, fluoranthene, pyrene, carbazole, and dibenzothiophene. Of the compounds studied, lignin peroxidase was able to oxidize those with ionization potentials of <8 eV (measured by electron impact). The reaction products contain hydroxyl and keto groups. In one case, carbon-carbon bond cleavage, yielding anthraquinone from 9-methylanthracene, was detected. Kinetic constants and stability characteristics of lignin peroxidase were determined by using pyrene as the substrate in systems containing different amounts of organic solvent. Benzyl alkylation of lignin peroxidase improved its activity in a system containing water-miscible organic solvent but did not increase its resistance to inactivation at high solvent concentrations. PMID- 16349177 TI - Mutations Affecting Hyphal Colonization and Pyoverdine Production in Pseudomonads Antagonistic toward Phytophthora parasitica. AB - In previous studies, Pseudomonas putida 06909 and Pseudomonas fluorescens 09906 suppressed populations of Phytophthora parasitica in the citrus rhizosphere, suggesting that these bacteria may be useful in biological control of citrus root rot. In this study we investigated the mechanisms of antagonism between the bacteria and the fungus. Both bacteria colonized Phytophthora hyphae and inhibited the fungus on agar media. A hyphal column assay was developed to measure the colonization of bacteria on fungal hyphae and to enrich for colonization-deficient mutants. In this way we identified Tn5 mutants of each pseudomonad that were not able to colonize the hyphae and inhibit fungal growth in vitro. Colonization-deficient mutants were nonmotile and lacked flagella. Survival of nonmotile mutants in a citrus soil was similar to survival of a random Tn5 mutant over a 52-day period. Additional screening of random Tn5 mutants of both pseudomonads for loss of fungal inhibition in vitro yielded two distinct types of mutants. Mutants of the first type were deficient in production of pyoverdines and in inhibition of the fungus in vitro, although they still colonized fungal hyphae. Mutants of the second type lacked flagella and were not able to colonize the hyphae or inhibit fungal growth. No role was found for antibiotic production by the two bacteria in the inhibition of the fungus. Our results suggest that both hyphal colonization and pyoverdine production are important in the inhibition of Phytophthora parasitica by P. fluorescens and P. putida in vitro. PMID- 16349178 TI - Production of Chitinases and beta-1,3-Glucanases by Stachybotrys elegans, a Mycoparasite of Rhizoctonia solani. AB - The in vitro production of chitinases and beta-1,3-glucanases by Stachybotrys elegans, a mycoparasite of Rhizoctonia solani, was examined under various culture conditions, such as carbon and nitrogen sources, pH, and incubation period. Production of both enzymes was influenced by the carbon source incorporated into the medium and was stimulated by acidic pH and NaNO(3). The activity of both enzymes was very low in culture filtrates from cells grown on glucose and sucrose compared with that detected on chitin (for chitinases) and cell wall fragments (for beta-1,3-glucanases). Protein electrophoresis revealed that, depending on the carbon source used, different isoforms of chitinases and beta-1,3-glucanases were detected. S. elegans culture filtrates, possessing beta-1,3-glucanase and chitinase activities, were capable of degrading R. solani mycelium. PMID- 16349179 TI - Integrative Cloning, Expression, and Stability of the cryIA(c) Gene from Bacillus thuringiensis subsp. kurstaki in a Recombinant Strain of Clavibacter xyli subsp. cynodontis. AB - A bacterial endophyte was engineered for insecticidal activity against the European corn borer. The cryIA(c) gene from Bacillus thuringiensis subsp. kurstaki was introduced into the chromosome of Clavibacter xyli subsp. cynodontis by using an integrative plasmid vector. The integration vectors pCG740 and pCG741 included the replicon pGEM5Zf(+), which is maintained in Escherichia coli but not in C. xyli subsp. cynodontis; tetM as a marker for selection in C. xyli subsp. cynodontis; and a chromosomal fragment of C. xyli subsp. cynodontis to allow for homologous recombination between the vector and the bacterial chromosome. Insertion of vector DNA into the chromosome was demonstrated by DNA hybridization. Recombinant strains MDR1.583 and MDR1.586 containing the cryIA(c) gene were shown to produce the 133,000-kDa protoxin and several smaller immunoreactive proteins. Both strains were equally toxic to insect larvae in bioassays. Significant insecticidal activity was demonstrated in planta. The cryIA(c) gene and the tetM gene introduced into strain MDR1.586 were shown to be deleted from some cells, thereby giving rise to a noninsecticidal segregant population. In DNA hybridization experiments and insect bioassays, these segregants were indistinguishable from the wild-type strain. Overall, these results demonstrate the plausibility of genetically engineered bacterial endophytes for insect control. PMID- 16349180 TI - Influence of the Gas-Water Interface on Transport of Microorganisms through Unsaturated Porous Media. AB - In this article, a new mechanism influencing the transport of microorganisms through unsaturated porous media is examined, and a new method for directly visualizing bacterial behavior within a porous medium under controlled chemical and flow conditions is introduced. Resting cells of hydrophilic and relatively hydrophobic bacterial strains isolated from groundwater were used as model microorganisms. The degree of hydrophobicity was determined by contact-angle measurements. Glass micromodels allowed the direct observation of bacterial behavior on a pore scale, and three types of sand columns with different gas saturations provided quantitative measurements of the observed phenomena on a porous medium scale. The reproducibility of each break-through curve was established in three to five repeated experiments. The data collected from the column experiments can be explained by phenomena directly observed in the micromodel experiments. The retention rate of bacteria is proportional to the gas saturation in porous media because of the preferential sorption of bacteria onto the gas-water interface over the solid-water interface. The degree of sorption is controlled mainly by cell surface hydrophobicity under the simulated groundwater conditions because of hydrophobic forces between the organisms and the interfaces. The sorption onto the gas-water interface is essentially irreversible because of capillary forces. This preferential and irreversible sorption at the gas-water interface strongly influences the movement and spatial distribution of microorganisms. PMID- 16349181 TI - Genetic and Biochemical Characterization of Nectria haematococca Strains with Adhesive and Adhesion-Reduced Macroconidia. AB - A previous study reported the isolation of two mutants (LE1 and LE2) of the plant pathogenic fungus Nectria haematococca (anamorph, Fusarium solani f. sp. cucurbitae) with macroconidia with reduced ability to adhere (Att) to zucchini fruits and polystyrene. The adhesion-reduced-phenotype in LE1 and LE2 macroconidia is temperature sensitive and dependent on the concentration of nutrients. Classical genetic analysis of progeny derived from LE1 identified a mutation in a genetic locus, named Att1. The 90-kDa glycoprotein and macroconidial tip mucilage which were previously associated with the development of adhesion competence in Att macroconidia are specifically associated with macroconidia; neither is produced on microconidia, which are relatively nonadherent. However, macroconidia of both Att and Att strains produce the 90-kDa glycoprotein and the macroconidial tip mucilage. PMID- 16349182 TI - Purification and Characterization of Aeromonas caviae ME-1 Xylanase V, Which Produces Exclusively Xylobiose from Xylan. AB - A xylanase, which produces exclusively xylobiose from oat spelt and birch xylans, was isolated from the culture medium of Aeromonas caviae ME-1. The enzyme (xylanase V) was purified by ammonium sulfate fractionation, hydrophobic interaction, and ion-exchange and gel filtration chromatographies. The homogeneity of the final preparation was demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and agarose gel electrofocusing. The molecular mass and isoelectric point of the xylanase were 46 kDa and 5.4, respectively. Xylanase V had a maximum activity at a pH of 6.8 and at a temperature between 30 and 37 degrees C. It was relatively stable at a pH between 5.0 and 8.6 and a temperature between 25 and 37 degrees C. When soluble birch xylan was used as the substrate, the enzyme had a K(m) and V(max) of 2 mg/ml and 182 mumol of xylose equivalent liberated . min . mg of protein, respectively. By the action of xylanase V on xylans (from oat spelt and birch), only one product corresponding to xylobiose was observed by thin-layer chromatography. The xylanase V putative product was confirmed to be xylobiose by acid and enzymatic hydrolyses. The xylanase had neither beta-xylosidase, alpha-l-arabinofuranosidase, cellulase, nor beta-1,3-xylanase activities. Xylotriose was the shortest substrate which the enzyme could attack. These findings suggest that xylanase V is a novel enzyme that cleaves a xylobiose unit from one of the ends of xylans, probably by an exomechanism. PMID- 16349183 TI - Mass Spectrometric Studies of the Effect of pH on the Accumulation of Intermediates in Denitrification by Paracoccus denitrificans. AB - We have used a quadrupole mass spectrometer with a gas-permeable membrane inlet for continuous measurements of the production of N(2)O and N(2) from nitrate or nitrite by cell suspensions of Paracoccus denitrificans. The use of nitrate and nitrite labeled with N was shown to simplify the interpretation of the results when these gases were measured. This approach was used to study the effect of pH on the production of denitrification intermediates from nitrate and nitrite under anoxic conditions. The kinetic patterns observed were quite different at acidic and alkaline pH values. At pH 5.5, first nitrate was converted to nitrite, then nitrite was converted to N(2)O, and finally N(2)O was converted to N(2). At pH 8.5, nitrate was converted directly to N(2), and the intermediates accumulated to only low steady-state concentrations. The sequential usage of nitrate, nitrite, and nitrous oxide observed at pH 5.5 was simulated by using a kinetic model of a branched electron transport chain in which alternative terminal reductases compete for a common reductant. PMID- 16349184 TI - Aerobic mineralization of trichloroethylene, vinyl chloride, and aromatic compounds by rhodococcus species. AB - Two Rhodococcus strains which were isolated from a trichloroethylene (TCE) degrading bacterial mixture and Rhodococcus rhodochrous ATCC 21197 mineralized vinyl chloride (VC) and TCE. Greater than 99.9% of a 1-mg/liter concentration of VC was degraded by cell suspensions. [1,2-C]VC was degraded by cell suspensions, with the production of greater than 66% CO(2) and 20% C-aqueous phase products and incorporation of 10% of the C into the biomass. Cultures that utilized propane as a substrate were able to mineralize greater than 28% of [1,2-C]TCE to CO(2), with approximately 40% appearing in C-aqueous phase products and another 10% of C incorporated into the biomass. VC degradation was oxygen dependent and occurred at a pH range of 5 to 10 and temperatures of 4 to 35 degrees C. Cell suspensions degraded up to 5 mg of TCE per liter and up to 40 mg of VC per liter. Propane competitively inhibited TCE degradation. Resting cell suspensions also degraded other chlorinated aliphatic hydrocarbons, such as chloroform, 1,1 dichloroethylene, and 1,1,1-trichloroethane. The isolates degraded a mixture of aromatic and chlorinated aliphatic solvents and utilized benzene, toluene, sodium benzoate, naphthalene, biphenyl, and n-alkanes ranging in size from propane to hexadecane as carbon and energy sources. The environmental isolates appeared more catabolically versatile than R. rhodochrous ATCC 21197. The data report that environmental isolates of Rhodococcus species and R. rhodochrous ATCC 21197 have the potential to degrade TCE and VC in addition to a variety of aromatic and chlorinated aliphatic compounds either individually or in mixtures. PMID- 16349185 TI - Mineral precipitation by epilithic biofilms in the speed river, ontario, Canada. AB - Epilithic microbial communities, ubiquitously found in biofilms on submerged granite, limestone, and sandstone, as well as on the concrete support pillars of bridges, were examined in the Speed River, Ontario, Canada. Transmission electron microscopy showed that attached bacteria (on all substrata) were highly mineralized, ranging from Fe-rich capsular material to fine-grained (<1 mum) authigenic (primary) mineral precipitates. The authigenic grains exhibited a wide range of morphologies, from amorphous gel-like phases to crystalline structures. Energy-dispersive X-ray spectroscopy indicated that the most abundant mineral associated with epilithic bacteria was a complex (Fe, Al) silicate of variable composition. The gel-like phases were similar in composition to a chamositic clay, whereas the crystalline structures were more siliceous and had compositions between those of glauconite and kaolinite. The consistent formation of (Fe, Al) silicates by all bacterial populations, regardless of substratum lithology, implies that biomineralization was a surface process associated with the anionic nature of the cell wall. The adsorption of dissolved constituents from the aqueous environment contributed significantly to the mineral formation process. In this regard, it appears that epilithic microbial biofilms dominate the reactivity of the rock-water interface and may determine the type of minerals formed, which will ultimately become part of the riverbed sediment. Because rivers typically contain high concentrations of dissolved iron, silicon, and aluminum, these findings provide a unique insight into biogeochemical activities that are potentially widespread in natural waters. PMID- 16349186 TI - Secretion of Ligninolytic Enzymes and Mineralization of C-Ring-Labelled Synthetic Lignin by Three Phlebia tremellosa Strains. AB - Production of ligninolytic enzymes by three strains of the white rot fungus Phlebia tremellosa (syn. Merulius tremellosus) was studied in bioreactor cultivation under nitrogen-limiting conditions. The Mn(II) concentration of the growth medium strongly affected the secretion patterns of lignin peroxidase and laccase. Two major lignin peroxidase isoenzymes were expressed in all strains. In addition, laccase and glyoxal oxidase were purified and characterized in one strain of P. tremellosa. In contrast, manganese peroxidase was not found in fast protein liquid chromatography profiles of extracellular proteins under either low (2.4 muM) or elevated (24 and 120 muM) Mn(II) concentrations. However, H(2)O(2)- and Mn-dependent phenol red-oxidizing activity was detected in cultures supplemented with higher Mn(II) levels. Mineralization rates of C-ring-labelled synthetic lignin (i.e., dehydrogenation polymerizate) by all strains under a low basal Mn(II) level were similar to those obtained for Phanerochaete chrysosporium and Phlebia radiata. A high manganese concentration repressed the evolution of CO(2) even when a chelating agent, sodium malonate, was included in the medium. PMID- 16349187 TI - Xylose and Glucose Utilization by Bacteroides xylanolyticus X5-1 Cells Grown in Batch and Continuous Culture. AB - During cultivation on a mixture of xylose and glucose, Bacteroides xylanolyticus X5-1 showed neither diauxic growth nor a substrate preference. Xylose-limited continuous-culture cells were able to consume xylose and glucose both as single substrates and as mixed substrates without any lag phase. When glucose was the growth-limiting substrate, the microorganism was unable to consume xylose. However, in the presence of a small amount of glucose or pyruvate, xylose was utilized after a short lag phase. In glucose-limited cells, xylose isomerase was present at low activity but xylulose kinase activity could not be detected. On addition of a mixture of xylose and glucose, xylose isomerase was induced immediately and xylulose kinase was induced after about 30 min. The induction of the two enzymes was sensitive to chloramphenicol, showing de novo synthesis. Xylose uptake in glucose-grown cells was very low, but the uptake rate could be increased when incubated with a xylose-glucose mixture. The increase in the uptake rate was not affected by chloramphenicol, indicating that a constitutive uptake system had to be activated. The inability of B. xylanolyticus X5-1 cells undergoing glucose-limited continuous culture to induce the xylose catabolic pathway after the addition of only xylose probably was caused by energy limitation. PMID- 16349188 TI - A Selective Medium for the Isolation and Quantification of Bradyrhizobium japonicum and Bradyrhizobium elkanii Strains from Soils and Inoculants. AB - The ecological examination of members of the family Rhizobiaceae has been hampered by the lack of a selective medium for isolation of root nodule bacteria from soil. A novel non-antibiotic-containing medium has been developed which allows selective isolation of Bradyrhizobium japonicum and B. elkanii strains from soil and inoculants. The medium, BJSM, is based on the resistance of B.japonicum and B. elkanii strains to more than 40 mug of the metals ions Zn and Co per ml. BJSM does not allow growth of Rhizobium sp. strains. We used BJSM to isolate bacteria from a Hubbard soil and from several commercially prepared soybean inoculants. Ninety-eight percent of the isolates obtained from Hubbard soil nodulated Glycine max cv. Kasota, and between 55 and 95% of the isolates from the commercial inoculants had the ability to nodulate soybeans. Numbers of bradyrhizobia obtained by using BJSM, strain-specific fluorescent antibodies, and the most-probable-number plant infection assay indicated that the three techniques were comparable in quantifying B. japonicum strains in soils and inoculants, although most-probable-number counts were generally 0.5 order of magnitude greater than those obtained by using BJSM. Results of our studies indicate that BJSM is useful for direct isolation and quantification of B. japonicum and B. elkanii from natural soils and inoculants. This medium may prove to be an important tool for autecological and enumeration studies of diverse populations of bradyrhizobia and as a quality control method for soybean inoculants. PMID- 16349189 TI - Physiological Aspects of Biosynthesis of Lignin Peroxidases by Phanerochaete chrysosporium. AB - Methods based on UV-visible diffuse reflectance spectroscopy were used to study the physiological aspects of lignin-peroxidase biosynthesis by Phanerochaete chrysosporium. Here we introduce the use of cytochrome aa(3) as an indicator of active fungal biomass and of its redox state to calculate the oxygen mass transport coefficient between the growth medium and the fungal cell interior. When lignin peroxidase biosynthesis was enhanced by the addition of Tween 80 or Tween 20 to the growth medium, a higher proportion of reduced cytochrome aa(3) and a higher oxygen diffusion barrier were observed compared with control cultures. In cultures supplemented with Tween 80 or Tween 20, a higher oxygen mass transport coefficient between the growth medium and the interior of the fungal cell was also found. The beginning of the lignin peroxidase activity in these cultures was found to coincide with a temporary cessation in the dry biomass increase and a reduction in the relative active-biomass concentration. During the lignin peroxidase activity, a decrease in the intracellular pH and an increase in the growth medium pH were determined in cultures supplemented with Tween 80. PMID- 16349190 TI - Purification and Characterization of an Inducible s-Triazine Hydrolase from Rhodococcus corallinus NRRL B-15444R. AB - The widespread use and relative persistence of s-triazine compounds such as atrazine and simazine have led to increasing concern about environmental contamination by these compounds. Few microbial isolates capable of transforming substituted s-triazines have been identified. Rhodococcus corallinus NRRL B-15444 has previously been shown to possess a hydrolase activity that is responsible for the dechlorination of the triazine compounds deethylsimazine (6-chloro-N-ethyl 1,3,5-triazine-2,4-diamine) (CEAT) and deethylatrazine (6-chloro-N-isopropyl 1,3,5-triazine-2,4-diamine) (CIAT). The enzyme responsible for this activity was purified and shown to be composed of four identical subunits of 54,000 Da. Kinetic experiments revealed that the purified enzyme is also capable of deaminating the structurally related s-triazine compounds melamine (2,4,6 triamino-1,3,5-triazine) (AAAT) and CAAT (2-chloro-4,6-diamino-1,3,5-triazine), as well as the pyrimidine compounds 2,4,6-triaminopyrimidine (AAAP) and 4-chloro 2,6-diaminopyrimidine (CAAP). The triazine herbicides atrazine and simazine inhibit the hydrolytic activities of the enzyme but are not substrates. Induction experiments demonstrate that triazine hydrolytic activity is inducible and that this activity rises approximately 20-fold during induction. PMID- 16349191 TI - Structure and composition of biological slimes on paper and board machines. AB - Biological slimes (biofilms) collected from the wet end of paper and board machines were examined by electron microscopy and analyzed for fatty acid composition, neutral sugar composition, and ATP. Electron microscopy revealed minuscule prokaryotic organisms (diameter, 0.2 to 0.4 mum). Larger cells morphologically resembling Sphaerotilus and Leptothrix spp. were found in slimes from machines using recycled fiber or unbleached pulp. The bacteria were embedded in a slimy matrix and often contained reserve materials microscopically resembling poly-beta-hydroxybutyrate and glycogen. Fatty acid analysis of the slimes revealed bacterial signature fatty acids in concentrations equivalent to the presence of 2 x 10 to 2.6 x 10 (average, 7 x 10) bacterial cells (live and dead) per g (dry weight) of slime. The slimes contained several known components of bacterial polysaccharides in addition to glucose, indicating that the slime body consisted of bacterial polysaccharides. The slimes contained uronic acids equivalent to a binding capacity of 12.5 to 50 mumol of divalent cations per g (dry weight) of slime. The uronic acid-containing polysaccharides may be responsible for the accumulation of heavy metals in the slime. Calculation of the ATP contents of the slimes resulted in an estimate of 5 x 10 cells per g (dry weight) of slime when calibrated with pure bacterial cultures isolated from the slimes. From electron micrographs, an estimate ranging from 1 x 10 to 1.5 x 10 (average, 4 x 10) cells per g (dry weight) of slime was obtained. PMID- 16349192 TI - Patterns of Reactivity between a Panel of Monoclonal Antibodies and Forage Rhizobium Strains. AB - A panel of 11 monoclonal antibodies raised against vegetative cells of Rhizobium leguminosarum biovar trifolii or Rhizobium meliloti was tested by enzyme-linked immunosorbent assay for reactivity with 47 strains of R. leguminosarum biovar trifolii and 60 strains of R. meliloti. The goal of the study was to define the degree of specificity associated with each antibody and to gain an understanding of the amount of antigenic diversity found among the strains and between the species. Each antibody was tested against each Rhizobium strain in four forms: washed steamed cells, washed unsteamed cells, cell-free culture broth, and nodule squash material. Each antibody showed a different pattern of reactivity among the 107 strains. One of each of the antibodies developed against R. meliloti and R. leguminosarum biovar trifolii reacted in a highly specific manner with cells or antigen from the immunogenic strain only. Nine of the antibodies recognized secreted as well as cellular antigen from many of the strains. Analysis of patterns of reactivity between the 107 strains and the 11 antibodies separated the strains into 28 groups of which 12 were represented by one strain only. PMID- 16349193 TI - Two Genomic Regions Involved in Catechol Siderophore Production by Erwinia carotovora. AB - Two regions involved in catechol biosynthesis (cbs) of Erwinia carotovora W3C105 were cloned by functional complementation of Escherichia coli mutants that were deficient in the biosynthesis of the catechol siderophore enterobactin (ent). A 4.3-kb region of genomic DNA of E. carotovora complemented the entB402 mutation of E. coli. A second genomic region of 12.8 kb complemented entD, entC147, entE405, and entA403 mutations of E. coli. Although functions encoded by catechol biosynthesis genes (cbsA, cbsB, cbsC, cbsD, and cbsE) of E. carotovora were interchangeable with those encoded by corresponding enterobactin biosynthesis genes (entA, entB, entC, entD, and entE), only cbsE hybridized to its functional counterpart (entE) in E. coli. The cbsEA region of E. carotovora W3C105 hybridized to genomic DNA of 21 diverse strains of E. carotovora but did not hybridize to that of a chrysobactin-producing strain of Erwinia chrysanthemi. Strains of E. carotovora fell into nine groups on the basis of sizes of restriction fragments that hybridized to the cbsEA region, indicating that catechol biosynthesis genes were highly polymorphic among strains of E. carotovora. PMID- 16349194 TI - Construction of a Bioinsecticidal Strain of Pseudomonas fluorescens Active against the Sugarcane Borer, Eldana saccharina. AB - A cryIA(c) gene was cloned from a native Bacillus thuringiensis strain showing activity against the sugarcane borer, Eldana saccharina. The sequence of the cloned gene was very similar to that of the B. thuringiensis subsp. kurstaki HD 73 cryIA(c) gene. The gene was introduced into an isolate of Pseudomonas fluorescens, capable of colonizing sugarcane, on two broad-host-range plasmids, pDER405 and pKT240, having copy numbers of 13 and 28, respectively. By using the Omegon-Km vector, the cry gene was introduced into the chromosome of P. fluorescens isolate 14. Bioassays on eldana larvae showed that the strain carrying the gene integrated into the chromosome was as toxic as one carrying it on pKT240. Glasshouse trials indicated that sugarcane treated with P. fluorescens 14::Omegon-Km-cry were more resistant to eldana damage than untreated sugarcane was. PMID- 16349195 TI - Recombination of the bph (Biphenyl) Catabolic Genes from Plasmid pWW100 and Their Deletion during Growth on Benzoate. AB - Pseudomonas sp. strain CB406 was isolated from polychlorinated biphenyl contaminated soil and harbors a nontransmissible plasmid, pWW100, of approximately 200 kb which carries the genes required for biphenyl and 4 chlorobiphenyl catabolism. The catabolic phenotype was mobilized following the construction in vivo of a cointegrate plasmid containing functional upper and lower biphenyl operons inserted into the broad-host-range R plasmid RP4. The Bph phenotype carried by pWW100 was stable in nonselective media but was unstable during growth on benzoate, where the sequential selection of two species of bph deletion derivatives occurs at high frequency. This mirrors observations made with TOL plasmids (encoding toluene and xylene catabolism) grown under similar conditions. Subcloning of dioxygenase genes involved in biphenyl catabolism confirmed the localization of the bph genes on the wild-type plasmid and the RP4 cointegrate plasmid. PMID- 16349196 TI - Biotransformation of the Herbicide Atrazine by the White Rot Fungus Phanerochaete chrysosporium. AB - Biotransformation of atrazine by the white rot fungus Phanerochaete chrysosporium was demonstrated by a 48% decrease of the initial herbicide concentration in the growth medium within the first 4 days of incubation, which corresponded to the mycelium-growing phase. Results clearly established the mineralization of the ethyl group of the herbicide. Analysis of the growth medium showed the formation of hydroxylated and/or N-dealkylated metabolites of atrazine during fungal degradation. PMID- 16349197 TI - Biosynthetic Pathway for Veratryl Alcohol in the Ligninolytic Fungus Phanerochaete chrysosporium. AB - Veratryl alcohol (VA) is a secondary metabolite of white-rot fungi that produce the ligninolytic enzyme lignin peroxidase. VA stabilizes lignin peroxidase, promotes the ability of this enzyme to oxidize a variety of physiological substrates, and is accordingly thought to play a significant role in fungal ligninolysis. Pulse-labeling and isotope-trapping experiments have now clarified the pathway for VA biosynthesis in the white-rot basidiomycete Phanerochaete chrysosporium. The pulse-labeling data, obtained with C-labeled phenylalanine, cinnamic acid, benzoic acid, and benzaldehyde, showed that radiocarbon labeling followed a reproducible sequence: it peaked first in cinnamate, then in benzoate and benzaldehyde, and finally in VA. Phenylalanine, cinnamate, benzoate, and benzaldehyde were all efficient precursors of VA in vivo. The isotope-trapping experiments showed that exogenous, unlabeled benzoate and benzaldehyde were effective traps of phenylalanine-derived C. These results support a pathway in which VA biosynthesis proceeds as follows: phenylalanine --> cinnamate --> benzoate and/or benzaldehyde --> VA. PMID- 16349198 TI - Stability and Activities of Antibiotics Produced during Infection of the Insect Galleria mellonella by Two Isolates of Xenorhabdus nematophilus. AB - Xenorhabdus nematophilus subsp. dutki, an entomopathogenic bacterium, is vectored by steinernematid nematodes into insects, where it produces broad-spectrum antibiotics. The use of the nematode-bacterium complex against soil-dwelling pest insects could introduce antibiotics into the soil via the dead insect fragments during the emergence phase of the nematodes. Studies on the stability and activities of these antibiotics produced in the insect Galleria mellonella may contribute to assessing the possible impact of antibiotics on soil bacteria. Two isolates of X. nematophilus subsp. dutki (isolates GI and SFU) produced xenocoumacins 1 and 2 in cadavers of G. mellonella larvae in a 1:1 ratio. Total xenocoumacin 1 and 2 production was 800 ng/200 mg (wet weight) of insect tissue for the GI isolate. Antibiotic activity of water extracts from insects that had been infected with X. nematophilus was stable at 60 degrees C for 1 h and after repeated freeze-thaw cycles. The antibiotic titer of extracts held at 27 degrees C declined by day 10. The spectrum of bacterial species killed by antibiotics produced in insect cadavers varied with the isolate of X. nematophilus. Levels of antibiotic activity were greater in vivo than in tryptic soy broth, which may represent a nutrient effect. The bacterial isolate, culture condition, and presence of nematodes influenced the total antibiotic production in vivo. However, the levels of activity were not correlated with bacterial levels in the different growth environments. Insect cadavers with antibiotic activity transiently lowered the numbers of the bacteria in the soil, the extent of decline varying with the strain of X. nematophilus and the time of sampling. PMID- 16349199 TI - Sensitivity of Iron-Oxidizing Bacteria, Thiobacillus ferrooxidans and Leptospirillum ferrooxidans, to Bisulfite Ion. AB - When grown on iron-salt medium supplemented with the bisulfite ion, Leptospirillum ferrooxidans was much more sensitive to the ion than was Thiobacillus ferrooxidans. The causes of the sensitivity of L. ferrooxidans to the bisulfite ion were studied. The bisulfite ion completely inhibited the iron oxidizing activities of L. ferrooxidans and T. ferrooxidans at 0.02 and 0.2 mM, respectively. A trapping reagent for the bisulfite ion, formaldehyde, completely reversed the inhibition. The treatment of intact cells with 1.0 mM bisulfite ion for 1 h and washing the bisulfite ion from the cells had no harmful effects on the iron-oxidizing activity of T. ferrooxidans. However, the treatment of L. ferrooxidans with 0.1 mM bisulfite ion for 1 h completely destroyed the iron oxidizing activity. T. ferrooxidans had sulfite:ferric ion oxidoreductase activity. In contrast, a quite low level of sulfite:ferric ion oxidoreductase activity was found in L. ferrooxidans, suggesting that it is much more difficult for L. ferrooxidans to oxidize the bisulfite ion to the less harmful sulfate than it is for T. ferrooxidans. These results suggest that the sensitivity of L. ferrooxidans to the bisulfite ion is due to a lack of an active sulfite:ferric ion oxidoreductase and the sensitivity of its iron oxidase to bisulfite ion. PMID- 16349200 TI - Reduction of Chromate by Desulfovibrio vulgaris and Its c(3) Cytochrome. AB - Washed cell suspensions of Desulfovibrio vulgaris rapidly reduced Cr(VI) to Cr(III) with H(2) as the electron donor. The c(3) cytochrome from this organism functioned as a Cr(VI) reductase. D. vulgaris may have advantages over previously described Cr(VI) reducers for the bioremediation of Cr(VI)-contaminated waters. PMID- 16349201 TI - Utilization of Glucose and Xylose in Ruminal Strains of Butyrivibrio fibrisolvens. AB - The dual-substrate utilization pattern in cultures of five ruminal strains of Butyrivibrio fibrisolvens growing on glucose and xylose was investigated. Strains ATCC 19171 and 86 utilized glucose and xylose simultaneously. Other strains exhibited diauxic growth. Strains X1 and CE 51 exhibited classical diauxic growth in which glucose was utilized during the first phase. Strain X2D62 displayed atypical diauxic growth in which slow utilization of xylose was followed by rapid utilization of glucose after the xylose depletion. The ATP-dependent phosphorylation of glucose was found in all strains tested. The phosphoenolpyruvate-dependent phosphorylation of glucose was detected only in B. fibrisolvens CE 51. PMID- 16349203 TI - A Simple RNA Probe System for Analysis of Listeria monocytogenes Polymerase Chain Reaction Products. AB - [This corrects the article on p. 2796 in vol. 59.]. PMID- 16349202 TI - A role for exopolysaccharides in the protection of microorganisms from desiccation. AB - Mucoid strains of Escherichia coli, Acinetobacter calcoaceticus, and Erwinia stewartii were significantly more resistant to desiccation than corresponding isogenic nonmucoid mutants (survival rates of up to 35% in mucoid strains and between 0.7 and 5% in nonmucoid variants), even in colonies containing both cell types. Desiccation was found to bring about an induction of beta-galactosidase in Lon strains of E. coli K-12 carrying transcriptional lac fusions in the capsule biosynthetic (cps) regulon. This induction was dependent on the transcriptional activators RcsA and RcsB. Induction was lower in cells carrying mutations in the membrane sensor protein RcsC. PMID- 16349205 TI - Quantitative reverse sample genome probing of microbial communities and its application to oil field production waters. AB - [This corrects the article on p. 4101 in vol. 59.]. PMID- 16349204 TI - Mobile bacteria and transport of polynuclear aromatic hydrocarbons in porous media. AB - [This corrects the article on p. 3309 in vol. 59.]. PMID- 16349206 TI - Biotinylation of Bacillus thuringiensis Insecticidal Crystal Proteins. AB - [This corrects the article on p. 1821 in vol. 59.]. PMID- 16349207 TI - Inactivation of ccmO in Synechococcus sp. Strain PCC 7942 Results in a Mutant Requiring High Levels of CO(2). AB - Inactivation of ccmO in Synechococcus sp. strain PCC 7942 resulted in a mutant which possesses aberrant carboxysomes and a normal inorganic carbon uptake capability but a reduced ability to photosynthetically utilize the internal inorganic carbon pool. Consequently, it exhibits low apparent photosynthetic affinity for extracellular inorganic carbon and demands high levels of CO(2) for growth. PMID- 16349208 TI - Evaluation of procedures for isolation of nontuberculous mycobacteria from soil and water. AB - Six methods of decontamination each for the isolation of mycobacteria from soil and water were compared. On the basis of the results obtained, three of the six methods for soil and two of the six methods for water were further evaluated. For both soil and water samples, the method using 3% sodium lauryl sulfate in combination with 1% NaOH yielded more positives than the other methods. PMID- 16349209 TI - Plasmid Transfer into the Homoacetogen Acetobacterium woodii by Electroporation and Conjugation. AB - Shuttle vectors (pMS3 and pMS4) which replicated in Escherichia coli and in gram positive Acetobacterium woodii were constructed by ligating the replication origin of plasmid pAMbeta1 with the E. coli cloning vector pUC19 and the tetM gene of streptococcal transposon Tn916. Electrotransformation of A. woodii was achieved at frequencies of 4.5 x 10 transformants per mug of plasmid DNA. For conjugal plasmid transfer, the mobilizable shuttle vector pKV12 was constructed by cloning the tetM gene into pAT187. Mating of E. coli containing pKV12 with A. woodii resulted in transfer frequencies of 3 x 10 to 7 x 10 per donor or recipient. PMID- 16349210 TI - Iron-Stimulated N(2) Fixation and Growth in Natural and Cultured Populations of the Planktonic Marine Cyanobacteria Trichodesmium spp. AB - In light of recent proposals that iron (Fe) availability may play an important role in controlling oceanic primary production and nutrient flux, its regulatory impact on N(2) fixation and production dynamics was investigated in the widespread and biogeochemically important diazotrophic, planktonic cyanobacteria Trichodesmium spp. Fe additions, as FeCl(3) and EDTA-chelated FeCl(3), enhanced N(2) fixation (nitrogenase activity), photosynthesis (CO(2) fixation), and growth (chlorophyll a production) in both naturally occurring and cultured (on unenriched oligotrophic seawater) Trichodesmium populations. Maximum enhancement of these processes occurred under FeEDTA-amended conditions. On occasions, EDTA alone led to enhancement. No evidence for previously proposed molybdenum or phosphorus limitation was found. Our findings geographically extend support for Fe limitation of N(2) fixation and primary production to tropical and subtropical oligotrophic ocean waters often characterized by Trichodesmium blooms. PMID- 16349211 TI - Fuel alcohol production: optimization of temperature for efficient very-high gravity fermentation. AB - The time required to end ferment wheat mash decreased as the temperature was increased from 17 to 33 degrees C, but it increased as the concentration of dissolved solids was raised from 14.0 to 36.5 g/100 ml. Ethanol yield was not appreciably affected. Over the range of fermentation temperatures tested, the addition of urea accelerated the rate of fermentation, decreased the time required to complete fermentation at all dissolved-solid concentrations, and stimulated the production of slightly more ethanol than was produced by the corresponding unsupplemented control mashes. The optimum temperature for maximum ethanol production in urea-supplemented very-high-gravity wheat mash was 27 degrees C. These data are important for the industrial assessment of very-high gravity fermentation technology. PMID- 16349212 TI - Bioavailability of chlorocatechols in naturally contaminated sediment samples and of chloroguaiacols covalently bound to c(2)-guaiacyl residues. AB - Bacteria in anaerobic enrichment cultures that dechlorinated a range of chlorocatechols were used to examine the stability of endogenous chlorocatechols in a contaminated sediment sample and in interstitial water prepared from it. During incubation of the sediment sample for 450 days with or without added cells, there was a decrease in the concentration of solvent-extractable chlorocatechols but not in that of the total chlorocatechols, including sediment associated components. In the presence of azide, the decrease in the concentrations of the former was eliminated or substantially decreased. Control experiments in which 3,4,5-trichlorocatechol was added to the sediment suspensions after 130 days showed that its dechlorination was accomplished not only by the added cells but also by the endemic microbial flora. It was concluded that the endogenous chlorocatechols in the sediment were not accessible to microorganisms with dechlorinating activity. On the other hand, microorganisms were apparently responsible for decreasing the solvent extractability of the chlorocatechols, and this effect decreased with increasing length of exposure time. Similar experiments carried out for 70 days with the sediment interstitial water showed that the chlorocatechols that were known to be associated with organic matter were also inaccessible to microbial dechlorination. Experiments with model compounds in which 4,5,6-trichloroguaiacol and tetrachloroguaiacol were covalently linked to C(2)-guaiacyl residues showed that these compounds were resistant to O demethylation or dechlorination during incubation with a culture having these activities. The only effect of microbial action was the quantitative reduction in 12 days of the C'1 keto group to an alcohol which was stable against further transformation for up to 65 days. The results of these experiments are consistent with the existence of chlorocatechols and chloroguaiacols in contaminated sediments and illustrate the cardinal significance of bioavailability in determining their recalcitrance to dechlorination and O demethylation, respectively. It is suggested that bioavailability is an important factor in determining the persistence of xenobiotics in natural ecosystems and that its omission represents a serious limitation in the interpretation of many laboratory experiments directed towards determining the persistence of xenobiotics in aquatic ecosystems. PMID- 16349213 TI - Production of Bacteriolytic Enzymes by Streptomyces globisporus Regulated by Exogenous Bacterial Cell Walls. AB - Mutanolysin biosynthesis and pigment production in Streptomyces globisporus ATCC 21553 were stimulated by adding bacterial cell walls to the medium. The increased bacteriolytic activity in the supernatant correlated with an increased de novo synthesis of mutanolysin and was between 4- and 20-fold higher than in cultures grown without bacterial cell walls. The increase in mutanolysin synthesis was brought about by enhanced transcription of the mutanolysin gene. The stimulation was only observed in medium which contained dextrin or starch as the carbon source. Glucose abolished the stimulation and also inhibited the low constitutive synthesis of mutanolysin. The induction of lytic activity was observed to require minimally 0.4 mg of bacterial cell walls per ml, whereas 0.6 mg of bacterial cell walls per ml yielded maximal lytic activity. Further supplements of bacterial cell walls did not result in enhanced lytic activity. The stimulation could be achieved independently of the phase of growth of the Streptomyces strain. Cultures grown in the presence of bacterial cell walls exhibited a higher growth yield. However, the accelerated growth was not the reason for the increased amount of mutanolysin produced. The growth of cultures with peptidoglycan monomers added to the medium instead of cell walls was similarly increased, but an effect on the biosynthesis of mutanolysin was not observed. All bacterial cell walls tested were capable of eliciting the stimulation of lytic activity, including cell walls of archaea, which contained pseudomurein. PMID- 16349214 TI - Characteristics and Diversity of beta-d-Glucosidase (EC 3.2.1.21) Activity in Marine Snow. AB - Large amorphous aggregates commonly described as marine snow were sampled in the water column of the northern Adriatic Sea in August 1991. beta-d-Glucosidase activity associated with these aggregates was characterized. Enzymatic activity was measured with the fluorogenic substrate analog 4-methylumbelliferyl-beta-d glucoside and found to be mainly associated with particles; on average, only 24% of the whole beta-d-glucosidase activity remained in the supernatant after centrifugation. Although the temperature optimum for beta-d-glucosidase activity was approximately 40 degrees C, incubation of the previously liberated particle attached enzyme at 50 degrees C for 20 min caused a >90% reduction of enzymatic activity relative to the activity at 40 degrees C. The level of inactivation of beta-d-glucosidase was much lower, however, when whole marine snow was incubated, indicating qualitative modifications of beta-d-glucosidase in marine snow. Separation of beta-d-glucosidase by different approaches indicated that the diversity of isoenzymes is restricted. In samples taken from the pycnocline, only one major isoenzyme was present in noticeable amounts. This isoenzyme contributed up to 70% of the whole beta-d-glucosidase activity detectable by two different chromatographic separations (anion-exchange chromatography and size exclusion chromatography). Although the same isoenzyme was dominant in marine snow taken from surface waters (0.5-m depth), we found a second peak of activity which eluted at lower NaCl concentrations from the anion-exchange column. Generally, the diversity of isoenzymes exhibiting beta-d-glucosidase activity seems to be surprisingly small in amorphous pelagic aggregates. PMID- 16349215 TI - Field evaluation of two colorimetric coliphage detection methods. AB - Two new methods for coliphage detection, a colorimetric agar-based (CAB) method and a liquid colorimetric presence-absence (LCPA) method, were compared to the coliphage method proposed by the American Public Health Association (APHA; Standard Methods for the Examination of Water and Wastewater, 18th ed., American Public Health Association, Washington, D.C., 1992). Both new methods are based on the induction of beta-galactosidase in Escherichia coli and the release of the enzyme through a lytic cell infection. The released enzyme then cleaves a chromogenic substrate which produces a colored reaction product. Ninety split water samples from four different sources were tested. A total of 52 samples were positive by the CAB method, 52 were positive by the LCPA method, and 53 were positive by the APHA method. Results indicated that (i) the CAB and LCPA methods were as sensitive in coliphage detection as the APHA method, (ii) both the CAB and LCPA methods were easier to read and interpret than the APHA method, and (iii) the CAB method detected more coliphages in a positive sample than the APHA method in two of the four types of water sources. Importantly, the rapid and simple LCPA method was as reliable and sensitive as either of the two agar-based methods in coliphage detection. PMID- 16349216 TI - Exogenous isolation of mobilizing plasmids from polluted soils and sludges. AB - Exogenous plasmid isolation was used to assess the presence of mobilizing plasmids in several soils and activated sludges. Triparental matings were performed with Escherichia coli (a member of the gamma subgroup of the Proteobacteria) as the donor of an IncQ plasmid (pMOL155, containing the heavy metal resistance genes czc: Co, Zn, and Cd), Alcaligenes eutrophus (a member of the beta subgroup of the Proteobacteria) as the recipient, and indigenous microorganisms from soil and sludge samples as helper strains. We developed an assay to assess the plasmid mobilization potential of a soil ecosystem on the basis of the number of transconjugants obtained after exogenous isolations. After inoculation into soil of several concentrations of a helper strain (E. coli CM120 harboring IncP [IncP1] mobilizing plasmid RP4), the log numbers of transconjugants obtained from exogenous isolations with different soil samples were a linear function of the log numbers of helper strain CM120(RP4) present in the soils. Four soils were analyzed for the presence of mobilizing elements, and mobilizing plasmids were isolated from two of these soils. Several sludge samples from different wastewater treatment plants yielded much higher numbers of transconjugants than the soil samples, indicating that higher numbers of mobilizing strains were present. The mobilizing plasmids isolated from Gent-O sludge and one plasmid isolated from Eislingen soil hybridized to the repP probe, whereas the plasmids isolated from Essen soil did not hybridize to a large number of rep probes (repFIC, repHI1, repH12, repL/M, repN, repP, repT, repU, repW, repX). This indicates that in Essen soil, broad-host-range mobilizing plasmids belonging to other incompatibility groups may be present. PMID- 16349217 TI - Impaction onto a Glass Slide or Agar versus Impingement into a Liquid for the Collection and Recovery of Airborne Microorganisms. AB - To study impaction versus impingement for the collection and recovery of viable airborne microorganisms, three new bioaerosol samplers have been designed and built. They differ from each other by the medium onto which the bioaerosol particles are collected (glass, agar, and liquid) but have the same inlet and collection geometries and the same sampling flow rate. The bioaerosol concentrations recorded by three different collection techniques have been compared with each other: impaction onto a glass slide, impaction onto an agar medium, and impingement into a liquid. It was found that the particle collection efficiency of agar slide impaction depends on the concentration of agar in the collection medium and on the sampling time, when samples are collected on a nonmoving agar slide. Impingement into a liquid showed anomalous behavior with respect to the sampling flow rate. Optimal sampling conditions in which all three new samplers exhibit the same overall sampling efficiency for nonbiological particles have been established. Inlet and collection efficiencies of about 100% have been achieved for all three devices at a sampling flow rate of 10 liters/min. The new agar slide impactor and the new impinger were then used to study the biological factors affecting the overall sampling efficiency. Laboratory experiments on the total recovery of a typical environmental microorganism, Pseudomonas fluorescens ATCC 13525, showed that both sampling methods, impaction and impingement, provided essentially the same total recovery when relatively nonstressed microorganisms were sampled under optimal sampling conditions. Comparison tests of the newly developed bioaerosol samplers with those commercially available showed that the incorporation of our research findings into the design of the new samplers yields better performance data than data from currently available samplers. PMID- 16349218 TI - Natural and Electroporation-Mediated Transformation of Methanococcus voltae Protoplasts. AB - The lack of high-efficiency transformation systems has severely impeded genetic research on methanogenic members of the kingdom Archaeobacteria. By using protoplasts of Methanococcus voltae and an integration vector, Mip1, previously shown to impart puromycin resistance, we obtained natural transformation frequencies that were about 80-fold higher (705 transformants per mug of transforming DNA) than that reported with whole cells. Electroporation-mediated transformation of M. voltae protoplasts with covalently closed circular Mip1 DNA was possible, but at lower frequencies of ca. 177 transformants per mug of vector DNA. However, a 380-fold improvement (3,417 transformants per mug of DNA) over the frequency of natural transformation with whole cells was achieved by electroporation of protoplasts with linearized DNA. This general approach, of using protoplasts, should allow the transformation of other methanogens, especially those that may be gently converted to protoplasts as a result of their tendency to lyse in hypotonic solutions. PMID- 16349219 TI - In vitro bleaching of hardwood kraft pulp by extracellular enzymes excreted from white rot fungi in a cultivation system using a membrane filter. AB - To clarify the role of excreted extracellular enzymes during long-term incubation in a pulp biobleaching system with white rot fungi, we developed a cultivation system in which a membrane filter is used; this membrane filter can prevent direct contact between hyphae and kraft pulp, but allows extracellular enzymes to attack the kraft pulp. Phanerochaete sordida YK-624 brightened the pulp 21.4 points to 54.0% brightness after a 5-day in vitro treatment; this value was significantly higher than the values obtained with Phanerochaete chrysosporium and Coriolus versicolor after a 7-day treatment. Our results indicate that cell free, membrane-filtered components from the in vitro bleaching system are capable of delignifying unbleached kraft pulp. Obvious candidates for filterable reagents capable of delignifying and bleaching kraft pulp are peroxidase and phenoloxidase proteins. The level of secreted manganese peroxidase activity in the filterable components was substantial during strain YK-624 in vitro bleaching. A positive correlation between the level of manganese peroxidase and brightening of the pulp was observed. PMID- 16349220 TI - Pressure stabilization of proteins from extreme thermophiles. AB - We describe the stabilization by pressure of enzymes, including a hydrogenase from Methanococcus jannaschii, an extremely thermophilic deep-sea methanogen. This is the first published report of proteins from thermophiles being stabilized by pressure. Inactivation studies of partially purified hydrogenases from an extreme thermophile (Methanococcus igneus), a moderate thermophile (Methanococcus thermolithotrophicus), and a mesophile (Methanococcus maripaludis), all from shallow marine sites, show that pressure stabilization is not unique to enzymes isolated from high-pressure environments. These studies suggest that pressure stabilization of an enzyme may be related to its thermophilicity. Further experiments comparing the effects of increased pressure on the stability of alpha glucosidases from the hyperthermophile Pyrococcus furiosus and Saccharomyces cerevisiae support this possibility. We have also examined pressure effects on several highly homologous glyceraldehyde-3-phosphate dehydrogenases from mesophilic and thermophilic sources and a rubredoxin from P. furiosus. The results suggest that hydrophobic interactions, which have been implicated in the stabilization of many thermophilic proteins, contribute to the pressure stabilization of enzymes from thermophiles. PMID- 16349221 TI - Phylogenetic Analysis of Bradyrhizobium japonicum and Photosynthetic Stem Nodulating Bacteria from Aeschynomene Species Grown in Separated Geographical Regions. AB - Nearly complete and short partial 16S rRNA sequences were derived from PCR amplified ribosomal DNAs of Bradyrhizobium japonicum USDA 136 and USDA 110 and five strains of bacteriochlorophyll-synthesizing bacteria isolated from stem nodules of Aeschynomene indica and other Aeschynomene species growing in different geographic regions, including India, The Philippines and North America. We confirmed that the five stem-nodulating strains examined synthesize bacteriochlorophyll a, and the absorption spectra of methanol-extracted cells contained a major absorbance peak at 770 nm. Strains isolated on different continents and from different Aeschynomene species were found to be phylogenetically homogeneous and exhibited levels of sequence similarity of more than 99%. The bacteriochlorophyll-synthesizing rhizobia, Bradyrhizobium japonicum, Blastobacter denitrificans, Afipia felis, and Rhodopseudomonas palustris exhibited levels of sequence similarity of 97% or greater and belong to a distinct line of descent within the alpha-2 subdivision of the Proteobacteria. Variable regions between positions 995 and 1045 provide potential target sites for design of a probe that is able to distinguish the photosynthetic rhizobia from closely related taxa. PMID- 16349222 TI - Changes in Cellular Fatty Acid Composition of Cephalosporium acremonium during Cephalosporin C Production. AB - Cephalosporium acremonium was cultivated in fermentation medium containing sucrose or methyl oleate as a carbon source for cephalosporin C production. The level of antibiotic production was 48 g of cephalosporin C per liter under optimum conditions when methyl oleate was used. The C(18:1) (oleic acid) methyl ester appeared to be utilized faster than the C(18:2) (linoleic acid) methyl ester in fermentation broth. Physiological characteristics of C. acremonium were investigated by determining the fatty acid composition of the total cellular free lipid. Significant changes in cellular fatty acid composition occurred during inoculum cultivation and fermentation. The percentage of C(18:1) increased from 19.1 to 38.5%, but the percentage of C(18:2) decreased from 56.7 to 36.1%, and there was an increase in pH during inoculum cultivation. The cellular fatty acid composition of C. acremonium grown in fermentation medium containing methyl oleate (methyl oleate medium) was significantly different from that in fermentation medium containing sucrose (sucrose medium). The major fatty acids detected were C(16:0) (palmitic acid), C(18:1), and C(18:2). In methyl oleate medium, the ratio of C(18:1) to C(18:2) increased from 0.34 to 1.37, while the cell morphology changed from hyphae to arthrospores and conidia. In contrast, in sucrose medium, the ratio of C(18:1) to C(18:2) decreased from 0.70 to 0.43, and most of the cells remained hyphal at the end of fermentation. We observed that hyphae contained a higher proportion of C(18:2) but arthrospores and conidia contained a higher proportion of C(18:1). PMID- 16349223 TI - Lignin-Degrading Enzymes of the Commercial Button Mushroom, Agaricus bisporus. AB - Agaricus bisporus, grown under standard composting conditions, was evaluated for its ability to produce lignin-degrading peroxidases, which have been shown to have an integral role in lignin degradation by wood-rotting fungi. The activity of manganese peroxidase was monitored throughout the production cycle of the fungus, from the time of colonization of the compost through the development of fruit bodies. Characterization of the enzyme was done with a crude compost extract. Manganese peroxidase was found to have a pI of 3.5 and a pH optimum of 5.4 to 5.5, with maximal activity during the initial stages of fruiting (pin stage). The activity declined considerably with fruit body maturation (first break). This apparent developmentally regulated pattern parallels that observed for laccase activity and for degradation of radiolabeled lignin and synthetic lignins by A. bisporus. Lignin peroxidase activity was not detected in the compost extracts. The correlation between the activities of manganese peroxidase and laccase and the degradation of lignin in A. bisporus suggests significant roles for these two enzymes in lignin degradation by this fungus. PMID- 16349224 TI - Isolation and Characterization of Sexual Spore Pigments from Aspergillus nidulans. AB - The homothallic ascomycete Aspergillus nidulans produces two types of pigmented spores: conidia and ascospores. The synthesis and localization of the spore pigments is developmentally regulated and occurs in specialized cell types. On the basis of spectroscopic evidence, we propose that the major ascospore pigment of A. nidulans (ascoquinone A) is a novel dimeric hydroxylated anthraquinone. The structure of ascoquinone A, as well as a comparison to model compounds, suggests that it is the product of a polyketide synthase. Previous studies have revealed that the conidial pigments from A. nidulans and a related Aspergillus species (A. parasiticus) also appear to be produced via polymerization of polyketide precursors (D. W. Brown, F. M. Hauser, R. Tommasi, S. Corlett, and J. J. Salvo, Tetrahedron Lett. 34:419-422, 1993; M. E. Mayorga and W. E. Timberlake, Mol. Gen. Genet. 235:205-212, 1992). The structural similarity between the ascospore pigment and the toxic anthraquinone norsolorinic acid, the first stable intermediate in the aflatoxin pathway, suggests an evolutionary relationship between the respective polyketide synthase systems. PMID- 16349225 TI - Phenotypic Consequences of Altering the Copy Number of abiA, a Gene Responsible for Aborting Bacteriophage Infections in Lactococcus lactis. AB - The abiA gene (formerly hsp) encodes an abortive phage infection mechanism which inhibits phage DNA replication. To analyze the effects of varying the abiA gene dosage on bacteriophage resistance in Lactococcus lactis, various genetic constructions were made. An IS946-based integration vector, pTRK75, was used to integrate a single copy of abiA into the chromosomes of two lactococcal strains, MG1363 and NCK203. In both strains, a single copy of abiA did not confer any significant phage resistance on the host except for one of the MG1363 integrants, NCK625, which exhibited a slightly higher level of resistance to phages sk1 and p2. Hybridization of the total cellular RNA from NCK625 to an abiA-specific probe indicated that the integration took place downstream of a promoter causing stronger expression of abiA in this integrant. Three abiA-containing plasmids of various copy numbers were introduced into both strains, and the recombinants were evaluated for resistance to phages c2, p2, sk1, and phi31. Plasmid pTRK18 has a copy number of approximately six (cn = 6) and caused a decreased plaque size for all phages evaluated. Integration of pTRK75 into a native plasmid of NCK203 generated pTRK362 (cn = 13), which caused a reduced efficiency of plaquing (EOP = 10) and reduced plaque size. A high-copy-number abiA plasmid (pTRK363), based on the pAMbeta1 origin of replication, was also constructed (cn = 100). Plasmid pTRK363 caused a significant reduction in EOP (10 to 10) and plaque size for all phages tested, although in some cases, this plasmid caused the evolution of AbiA resistant phage derivatives. Altering the gene dosage or expression level of abiA significantly affects the phage resistance levels. PMID- 16349226 TI - Variation of clonal, mesquite-associated rhizobial and bradyrhizobial populations from surface and deep soils by symbiotic gene region restriction fragment length polymorphism and plasmid profile analysis. AB - Genetic characteristics of 14 Rhizobium and 9 Bradyrhizobium mesquite (Prosopis glandulosa)-nodulating strains isolated from surface (0- to 0.5-m) and deep (4- to 6-m) rooting zones were determined in order to examine the hypothesis that surface- and deep-soil symbiont populations were related but had become genetically distinct during adaptation to contrasting soil conditions. To examine genetic diversity, Southern blots of PstI-digested genomic DNA were sequentially hybridized with the nodDABC region of Rhizobium meliloti, the Klebsiella pneumoniae nifHDK region encoding nitrogenase structural genes, and the chromosome-localized ndvB region of R. meliloti. Plasmid profile and host plant nodulation assays were also made. Isolates from mesquite nodulated beans and cowpeas but not alfalfa, clover, or soybeans. Mesquite was nodulated by diverse species of symbionts (R. meliloti, Rhizobium leguminosarum bv. phaseoli, and Parasponia bradyrhizobia). There were no differences within the groups of mesquite-associated rhizobia or bradyrhizobia in cross-inoculation response. The ndvB hybridization results showed the greatest genetic diversity among rhizobial strains. The pattern of ndvB-hybridizing fragments suggested that surface and deep strains were clonally related, but groups of related strains from each soil depth could be distinguished. Less variation was found with nifHDK and nodDABC probes. Large plasmids (>1,500 kb) were observed in all rhizobia and some bradyrhizobia. Profiles of plasmids of less than 1,000 kb were related to the soil depth and the genus of the symbiont. We suggest that interacting selection pressures for symbiotic competence and free-living survival, coupled with soil conditions that restrict genetic exchange between surface and deep-soil populations, led to the observed patterns of genetic diversity. PMID- 16349227 TI - Relative role of eukaryotic and prokaryotic microorganisms in phenanthrene transformation in coastal sediments. AB - THE RELATIVE ROLE OF EUKARYOTIC VERSUS PROKARYOTIC MICROORGANISMS IN PHENANTHRENE TRANSFORMATION WAS MEASURED IN SLURRIES OF COASTAL SEDIMENT BY TWO DIFFERENT APPROACHES: detection of marker metabolites and use of selective inhibitors on phenanthrene biotransformation. Phenanthrene biotransformation was measured by polar metabolite formation and CO(2) evolution from [9-C]phenanthrene. Radiolabeled metabolites were tentatively identified by high-performance liquid chromatography (HPLC) separation combined with UV/visible spectral analysis of HPLC peaks and comparison to authentic standards. Both yeasts and bacteria transformed phenanthrene in slurries of coastal sediment. Two products of phenanthrene oxidation by fungi, phenanthrene trans-3,4-dihydrodiol and 3 phenanthrol, were produced in yeast-inoculated sterile sediment. However, only products of phenanthrene oxidation typical of bacterial transformation, 1-hydroxy 2-naphthoic acid and phenanthrene cis-3,4-dihydrodiol, were isolated from slurries of coastal sediment with natural microbial populations. Phenanthrene trans-dihydrodiols or other products of fungal oxidation of phenanthrene were not detected in the slurry containing a natural microbial population. A predominant role for bacterial transformation of phenanthrene was also suggested from selective inhibitor experiments. Addition of streptomycin to slurries, at a concentration which suppressed bacterial viable counts and rates of [methyl H]thymidine uptake, completely inhibited phenanthrene transformation. Treatment with colchicine, at a concentration which suppressed yeast viable counts, depressed phenanthrene transformation by 40%, and this was likely due to nontarget inhibition of bacterial activity. The relative contribution of eukaryotic microorganisms to phenanthrene transformation in inoculated sterile sediment was estimated to be less than 3% of the total activity. We conclude that the predominant degraders of phenanthrene in muddy coastal sediments are bacteria and not eukaryotic microorganisms. PMID- 16349228 TI - Determination of diffusion coefficients in biofilms by confocal laser microscopy. AB - Microbial exopolymer may hinder the diffusion of nutrients, antibiotics, and other materials to the cell surface. Studies of diffusion in biofilms have been limited to indirect measurements. This study demonstrated the use of fluorescein and size-fractionated fluor-conjugated dextrans in conjunction with scanning confocal laser microscopy to directly monitor and determine diffusion coefficients within biofilms. The monitoring approaches were simple and, when combined with computerized image collection, allowed assembly of a data set suitable for calculation of one-dimensional diffusion coefficients for biofilm regions. With these techniques, it was shown that regional variability in the mobility of the dextrans occurred within mixed-species biofilms. Some regions exhibited rapid diffusion of all test molecules, while adjacent regions were only penetrated by the lower-molecular-weight compounds. The effective diffusion coefficients (D(e)) determined in a mixed-species biofilm were a function of the molecular radius of the probe (i.e., fluorescein, D(e) = 7.7 x 10 cm s; 4,000 molecular weight, D(e) = 3.1 x 10 cm s; and 2,000,000 molecular weight, D(e) = 0.7 x 10 cm s). These results demonstrated that diffusion in the biofilm was hindered relative to diffusion in the bulk solution. The study indicated that in situ monitoring by scanning laser microscopy is a useful approach for determining the mobility of fluorescently labeled molecules in biofilms, allowing image acquisition, appropriate scales of study, both xy and xz monitoring, and calculation of D(e) values. PMID- 16349229 TI - Effects of temperature stress on bean-nodulating Rhizobium strains. AB - High soil temperatures in tropical areas limit nodulation and dinitrogen fixation by strains of Rhizobium. Several heat-tolerant bean-nodulating Rhizobium strains have been isolated previously. However, the basis of their resistance to heat remains unknown. In this study, we compared the effects of heat on symbiotic nitrogen fixation, cell survival, amino acid uptake, and protein synthesis in a heat-tolerant (CIAT899) and a heat-sensitive (CNPAF512) bean-nodulating Rhizobium strain. Acetylene reduction activity of nodulated roots excised from unstressed plants was strongly diminished at 35 or 40 degrees C when plants were nodulated either by CIAT899 or by CNPAF512. When these strains were tested under free living conditions, survival at 40 degrees C as well as the kinetics of l [S]methionine uptake and protein synthesis at 35 and 40 degrees C indicated the higher tolerance of CIAT899 than of CNPAF512 to thermal stress. The synthesis of heat shock proteins was detected in both strains, although at different temperatures. Increased synthesis of 14 heat shock proteins in CNPAF512 and of 6 heat shock proteins in CIAT899 was observed at 40 and 45 degrees C, respectively. A heat shock protein of approximately 21 kDa, of which the synthesis was strongest in both Rhizobium strains upon a temperature shift up, was also conserved in several other bean-nodulating rhizobia. Acquired thermotolerance in CIAT899 was shown to depend on protein synthesis. PMID- 16349230 TI - Generation and Characterization of Environmentally Sensitive Variants of the beta Galactosidase from Lactobacillus delbrueckii subsp. bulgaricus. AB - A method is described for generating and screening variants of the beta galactosidase from Lactobacillus delbrueckii subsp. bulgaricus sensitive to several environmental stresses, with potential application in the food industry. Chemical mutagenesis with hydroxylamine or methoxylamine was performed on the beta-galactosidase gene carried on an Escherichia coli expression vector. Mutants sensitive to cold, heat, low pH, low magnesium concentration, and the presence of urea were isolated by screening for reduced color development on beta galactosidase indicator plates. The mutations responsible for three variant beta galactosidases were localized, and the base substitutions were determined by DNA sequencing. The amino acid alterations associated with one low-pH-sensitive (pHs) and two urea-sensitive (Us) variants correspond to P584L (pHs1), G400S/R479Q (Us26), and G167E/E168K/E363K/V492M (Us17), respectively. Mutant pHs1 is also heat, cold, low magnesium, and urea sensitive; Us26 is also cold sensitive; and Us17 is also low-pH sensitive. PMID- 16349231 TI - Archaeoglobus fulgidus Isolated from Hot North Sea Oil Field Waters. AB - A hyperthermophilic sulfate reducer, strain 7324, was isolated from hot (75 degrees C) oil field waters from an oil production platform in the Norwegian sector of the North Sea. It was enriched on a complex medium and isolated on lactate with sulfate. The cells were nonmotile, irregular coccoid to disc shaped, and 0.3 to 1.0 mum wide. The temperature for growth was between 60 and 85 degrees C with an optimum of 76 degrees C. Lactate, pyruvate, and valerate plus H(2) were utilized as carbon and energy sources with sulfate as electron acceptor. Lactate was completely oxidized to CO(2). The cells contained an active carbon monoxide dehydrogenase but no 2-oxoglutarate dehydrogenase activity, indicating that lactate was oxidized to CO(2) via the acetyl coenzyme A/carbon monoxide dehydrogenase pathway. The cells produced small amounts of methane simultaneously with sulfate reduction. F(420) was detected in the cells which showed a blue green fluorescence at 420 nm. On the basis of morphological, physiological, and serological features, the isolate was classified as an Archaeoglobus sp. Strain 7324 showed 100% DNA-DNA homology with A. fulgidus Z, indicating that it belongs to the species A. fulgidus. Archaeoglobus sp. has been selectively enriched and immunomagnetically captured from oil field waters from three different platforms in the North Sea. Our results show that strain 7324 may grow in oil reservoirs at 70 to 85 degrees C and contribute to hydrogen sulfide formation in this environment. PMID- 16349232 TI - Identification, Biosynthesis, and Function of 1,3,4,6-Hexanetetracarboxylic Acid in Methanobacterium thermoautotrophicum DeltaH. AB - An unusual compound, 1,3,4,6-hexanetetracarboxylic acid, was identified by H and C two-dimensional nuclear magnetic resonance spectroscopy and gas chromatography mass spectrometry as one of the major components of the small-molecule pool in Methanobacterium thermoautotrophicum DeltaH under optimal conditions of cell growth. Incorporation of C- and H-labeled acetates was consistent with the biosynthesis of this tetracarboxylic acid from alpha-ketoglutarate, two molecules of acetyl-coenzyme A, and one molecule of CO(2), as established for the tetracarboxylic acid moiety of methanofuran. CO(2) pulse- CO(2) chase methodology was used to establish the turnover rate for this compound. In contrast to the two other major solutes in this bacterium, cyclic 2,3-diphosphoglycerate and glutamate, which are key metabolic intermediates, this free tetracarboxylic acid was metabolically inactive, with a half-life that exceeded the cell doubling time. Hence, this molecular pool cannot serve as a metabolic intermediate in cell biosynthesis. The functional role of free tetracarboxylate as a conservative part of a system that maintains high positive internal osmotic pressure in this bacterium is proposed. PMID- 16349233 TI - Galactose Expulsion during Lactose Metabolism in Lactococcus lactis subsp. cremoris FD1 Due to Dephosphorylation of Intracellular Galactose 6-Phosphate. AB - In Lactococcus lactis subsp. cremoris FD1, galactose and lactose are both transported and phosphorylated by phosphotransferase systems. Lactose 6-phosphate (lactose-6P) is hydrolyzed intracellularly to galactose-6P and glucose. Glucose enters glycolysis as glucose-6P, whereas galactose-6P is metabolized via the tagatose-6P pathway and enters glycolysis at the tagatose diphosphate and fructose diphosphate pool. Galactose would therefore be a gluconeogenic sugar in L. lactis subsp. cremoris FD1, but since fructose 1,6-diphosphatase is not present in this strain, galactose cannot serve as an essential biomass precursor (glucose-6P or fructose-6P) but only as an energy (ATP) source. Analysis of the growth energetics shows that transition from N limitation to limitation by glucose-6P or fructose-6P gives rise to a very high growth-related ATP consumption (152 mmol of ATP per g of biomass) compared with the value in cultures which are not limited by glucose-6P or fructose-6P (15 to 50 mmol of ATP per g of biomass). During lactose metabolism, the galactose flux through the tagatose-6P pathway (r(max) = 1.2 h) is lower than the glucose flux through glycolysis (r(max) = 1.5 h) and intracellular galactose-6P is dephosphorylated; this is followed by expulsion of galactose. Expulsion of a metabolizable sugar has not been reported previously, and the specific rate of galactose expulsion is up to 0.61 g of galactose g of biomass h depending on the lactose flux and the metabolic state of the bacteria. Galactose excreted during batch fermentation on lactose is reabsorbed and metabolized when lactose is depleted from the medium. In vitro incubation of galactose-6P (50 mM) and permeabilized cells (8 g/liter) gives a supernatant containing free galactose (50 mM) but no P(i) (less than 0.5 mM). No organic compound except the liberated galactose is present in sufficient concentration to bind the phosphate. Phosphate is quantitatively recovered in the supernatant as P(i) by hydrolysis with alkaline phosphatase (EC 3.1.3.1), whereas inorganic pyrophosphatase (EC 3.6.1.1) cannot hydrolyze the compound. The results indicate that the unknown phosphate-containing compound might be polyphosphate. PMID- 16349234 TI - Genetic Structure of Rhizobium etli biovar phaseoli Associated with Wild and Cultivated Bean Plants (Phaseolus vulgaris and Phaseolus coccineus) in Morelos, Mexico. AB - The genetic structure of Rhizobium etli biovar phaseoli was determined for five populations in three different locations in the state of Morelos, Mexico, by using starch gel electrophoresis for five to nine polymorphic loci. Two populations were sampled during two different years from nodules of cultivated and wild common bean plants (Phaseolus vulgaris). The three other populations were associated with wild runner beans (P. coccineus) and sampled during 1988. The Rhizobium populations differ genetically both among sites and among populations within the same site in different years, as shown by differences in allelic frequencies, genetic differentiation analysis, and differences in electrotypes. The total genetic diversity for the five populations during 1988 was H = 0.487; there were also high levels of genetic variation within each population. We found the highest linkage disequilibrium in a global analysis for all the populations. At a local scale, we also found significant linkage disequilibrium in two populations, although the distribution of the D' suggest some recombination at a local scale. The other three rhizobium populations exhibit low linkage disequilibrium. A cluster analysis (UPGMA) of pairwise genetic distances showed that bacteria isolated from most wild Phaseolus spp. are grouped by population, whereas those obtained from cultivated P. vulgaris are very heterogeneous. The analysis of the genetic structure of Rhizobium strains may allow the identification of strains that are naturally well adapted to a wide range of different environments, which may be useful for agricultural purposes or as a starting point for developing improved Rhizobium strains. PMID- 16349235 TI - A diffusion gradient chamber for studying microbial behavior and separating microorganisms. AB - The natural habitats of most microbes are dynamic and include spatial gradients of growth substrates, electron acceptors, pH, salts, and inhibitory compounds. To mimic this diffusive aspect of nature, we developed an analytical diffusion gradient chamber (DGC) that can be used to separate, enrich for, isolate, and study the behavior of microorganisms. The chamber is a polycarbonate box containing an arena (5 by 5 by 2 cm) into which is cast a semisolid growth medium. Continuously replenished solute reservoirs positioned on each side of the arena but separated from it by a porous membrane enable the formation throughout the gel of multiple, intersecting gradients of solutes in two dimensions. With glucose as the solute, a gradient which spanned a 100-fold range in concentration was established across the arena in about 4 days. The shape of the glucose gradient was accurately predicted by a mathematical model based on Fickian diffusion. The growth and migratory behavior of Escherichia coli in response to imposed gradients of attractants (aspartate, alpha-methyl aspartate, and serine) and a repellent (valine) were examined. Cells responded in predictable ways to such gradients by forming distinctive growth and migration patterns in the DGC. This was true for wild-type E. coli as well as specific chemotaxis and motility mutants. The patterns yielded information about the threshold concentration of chemoeffectors needed to elicit a response as well as their saturating concentration. It was also evident that the metabolism of attractants significantly affected the gradients and, hence, the movement of cells. Finally, it was possible to separate E. coli and Pseudomonas fluorescens in the DGC on the basis of their differential responses to gradients of various chemoeffectors. PMID- 16349236 TI - Nutrient effect on the biological leaching of a black-schist ore. AB - The purpose of the study was to examine the influence of inorganic N (NH(4), NO(3)) and phosphate on the biological oxidation of a sulfidic black-schist ore which contained pyrrhotite as the main iron sulfide. Iron was initially solubilized as Fe from the ore and subsequently oxidized to Fe in shake flask experiments. Under these experimental conditions, iron dissolution from pyrrhotite was mainly a chemical reaction, with some enhancement by bacteria, whereas the subsequent Fe oxidation was bacterially mediated, with negligible contribution from chemical oxidation. Phosphate amendment did not enhance Fe oxidation. Chemical analysis of leach solutions with no exogenous phosphate revealed that phosphate was solubilized from the black-schist ore. Ammonium amendment (6 mM) enhanced Fe oxidation, whereas the addition of nitrate (6 and 12 mM) had a negative effect. An increase in the temperature from 30 to 35 degrees C slightly enhanced Fe oxidation, but the effect was statistically not significant. The precipitation of potassium jarosite was indicative of Fe oxidation and was absent in nitrate-inhibited cultures because of the lack of Fe oxidation. The black-schist ore also contained phlogopite, which was altered to vermiculite in iron-oxidizing cultures. PMID- 16349237 TI - Microbial transformation of Ibuprofen by a nocardia species. AB - The carboxylic acid functional group of ibuprofen [alpha-methyl-4-(2 methylpropyl) benzene acetic acid] is reduced to the corresponding alcohol and subsequently esterified to the acetate derivative by cultures of Nocardia species strain NRRL 5646. The alcohol and ester microbial transformation products were isolated, and their structures were determined by H and C nuclear magnetic resonance spectroscopy and mass spectrometry. By derivatization of synthetic and microbiologically produced ibuprofen alcohols with S(+)-O-acetylmandelic acid, nuclear magnetic resonance analysis indicated that the carboxylic acid reductase of Nocardia sp. is R enantioselective, giving alcohol products with an enantiomeric excess of 61.2%. The R enantioselectivity of the carboxylic acid reductase enzyme system was confirmed by using cell extracts together with ATP and NADPH in the reduction of isomeric ibuprofens. PMID- 16349238 TI - Effects of temperature, sulfide, and food abundance on growth and feeding of anaerobic ciliates. AB - The trophic role of ciliates in anaerobic food webs has not been assessed experimentally. In order to obtain basic information necessary to interpret field situations, we studied the effects of temperature, sulfide concentration, and food abundance on the growth and feeding activities of two anaerobic ciliates, Plagiopyla nasuta and Metopus es. The growth rate of P. nasuta increased with temperature from 8 to 18 degrees C (Q(10) = 2.0) and remained constant in the range between 18 and 24 degrees C (0.22 day). Sulfide concentrations of between 0 and 1 mM did not affect the feeding activities, but concentrations greater than 2 mM were inhibitory. The functional response of P. nasuta feeding on fluorescently labeled heterotrophic and phototrophic bacteria was investigated. In both cases, the parameters of the functional response were almost identical when expressed in terms of biovolume: the maximal uptake rate (U(m)) was 1,800 mum ciliate h and the half-saturation constant for ingestion (k) was 1.5 x 10 mum ml. The functional response of M. es feeding on heterotrophic bacteria was found to be similar to that of P. nasuta. These ciliates needed high bacterial abundances in order to maintain their growth (k of about 4 x 10 bacteria ml), implying that they will frequently be food limited in planktonic environments. Both the maximal uptake rates and the maximal clearance rates were comparable to those of aerobic ciliates. By combining the growth and feeding data, we estimated gross growth efficiencies of 12 and 13% for P. nasuta and M. es, respectively. These results indicate that the feeding rates of anaerobic ciliates are similar to those of aerobic ciliates. Their slower growth must, therefore, be due to the lower gross growth efficiency (likely due to anaerobic metabolism). PMID- 16349239 TI - Role of anaerobic ciliates in planktonic food webs: abundance, feeding, and impact on bacteria in the field. AB - We studied the dynamics of two populations of anaerobic ciliates, Plagiopyla sp. and Metopus sp., and of their potential prey, heterotrophic and phototrophic purple bacteria, in Lake Ciso throughout a 1-year cycle. The abundance of both ciliates was very low (less than 2 individuals per ml). During mixing, Plagiopyla ciliates exhibited high clearance rates (about 100 nl ciliate h), its integrated abundance increased with a net doubling time of 47 days, and its potential doubling times, as calculated from the number of bacteria consumed, ranged between 5 and 8 days. During stratification, the activity of Plagiopyla ciliates was reduced and the population decreased; this was related to the higher amounts of sulfide present. The impact of predation by the Plagiopyla population on bacterioplankton was found to be insignificant, less than 0.1% of bacterial biomass consumed per day. Thus, anaerobic ciliates cannot control the bacterioplankton in Lake Ciso because of both the low abundance over the period studied and the low feeding rates during certain periods. A review of available field studies suggests that this conclusion can be extrapolated to most other anoxic systems. PMID- 16349240 TI - Regulation of Product Formation in Bacteroides xylanolyticus X5-1 by Interspecies Electron Transfer. AB - Bacteroides xylanolyticus X5-1 was grown in pure culture and in mixed culture with Methanospirillum hungatei JF-1 under xylose limitation in the chemostat. In the pure culture, ethanol, acetate, CO(2), and hydrogen were the products. In the mixed culture, acetate, CO(2), and presumably hydrogen were the only products formed by B. xylanolyticus X5-1. The biomass yield of B. xylanolyticus X5-1 increased because of cocultivation. In cell extracts of the pure culture, both NAD- and NADP-dependent acetaldehyde dehydrogenase and ethanol dehydrogenase activities were found. In cell extracts of the mixed culture, activities of these enzymes were not detected. Inhibition of methanogenesis in the mixed culture by the addition of bromoethanosulfonic acid (BES) resulted in an accumulation of H(2), ethanol, and formate. Immediately after the addition of BES, NAD-dependent acetaldehyde dehydrogenase and ethanol dehydrogenase activities were detected. After a short lag phase, a NADP-dependent ethanol dehydrogenase was also detectable. The induction of acetaldehyde dehydrogenase and ethanol dehydrogenase was inhibited by chloramphenicol, suggesting de novo synthesis of these enzymes. These results are consistent with a model in which the shift in product formation caused by interspecies electron transfer is regulated at the level of enzyme synthesis. PMID- 16349241 TI - Composition and Rheological Properties of Extracellular Polysaccharide 105-4 Produced by Pseudomonas sp. Strain ATCC 53923. AB - A soil isolate produced a novel extracellular polysaccharide (EPS) with unusually potent thickening powers. The EPS contained d-mannose, d-glucose, d-galactose, and d-glucuronic acid in the unique molar ratio 1:4:1:2 and 10 to 15% acetate. Viscosities of a 1-g/liter aqueous solution were 1 x 10 and 14 x 10 cP at shear rates of 0.01 and 0.1 s, respectively. The EPS was insensitive to high concentrations of NaCl and CaCl(2). PMID- 16349242 TI - Optimization of Novel Extracellular Polysaccharide Production by an Enterobacter sp, on Wood Hydrolysates. AB - An environmental isolate identified as Enterobacter cloacae has been found to produce a highly viscous, anionic extracellular polysaccharide (EPS) from a weak mineral acid hydrolysate of hardwood. Production of this EPS has been optimized on the hydrolysate (initial pH, 6.3; NH(4)Cl amendment, 0.1%) so that crude yields approaching 9.83 g/liter were obtained. Although this EPS is polydisperse, its molecular mass as determined by gel exclusion chromatography centers at approximately 1,700 kDa. Solutions of this EPS have been examined rheologically under a variety of conditions and compare favorably with both xanthan and alginate. PMID- 16349243 TI - Acetate synthesis in soil from a bavarian beech forest. AB - Soil obtained from a beech forest formed significant amounts of acetate when incubated in a bicarbonate-buffered, mineral salt solution under anaerobic conditions at both 5 and 20 degrees C (21 and 38 g of acetate per kg [dry weight] of soil, respectively). At 20 degrees C, following an 18-day lag period, rates of 0.07 mmol of acetate synthesized per g (dry weight) of soil per day were observed. Acetate was not subject to immediate turnover; methane and hydrogen were not formed during the time intervals (5 degrees C, 335 days; 20 degrees C, 95 days) evaluated. The synthesis of acetate from endogenous materials was coincident with acetogenic potentials, i.e., the capacity to catalyze the H(2) dependent synthesis of acetate. Hydrogen consumption was not directed towards the synthesis of methane. Collectively, these results suggest that acetogenesis may be an underlying microbial activity of this forest soil. PMID- 16349244 TI - Effects of Sainfoin (Onobrychis viciifolia Scop.) Condensed Tannins on Growth and Proteolysis by Four Strains of Ruminal Bacteria. AB - Sainfoin leaf condensed tannins inhibited growth and protease activity in Butyrivibrio fibrisolvens A38 and Streptococcus bovis 45S1 but had little effect on Prevotella ruminicola B(1)4 or Ruminobacter amylophilus WP225. Tannins bound to cell coat polymers in all strains. Morphological changes in B. fibrisolvens and S. bovis implicated the cell wall as a target of tannin toxicity. PMID- 16349245 TI - A simple method that uses differential staining and light microscopy to assess the selectivity of wood delignification by white rot fungi. AB - CRYOSTAT MICROTOME SECTIONS OF BIRCH WOOD DEGRADED BY WHITE ROT FUNGI WERE EXAMINED BY LIGHT MICROSCOPY AFTER TREATMENT WITH TWO STAINS: astra-blue, which stains cellulose blue only in the absence of lignin, and safranin, which stains lignin regardless of whether cellulose is present. The method provided a simple and reliable screening procedure that distinguishes between fungi that cause decay by selectively removing lignin and those that degrade both cellulose and lignin simultaneously. Moreover, morphological characteristics specific to selective delignification were revealed. PMID- 16349246 TI - Effect of Residual Lignin Type and Amount on Bleaching of Kraft Pulp by Trametes versicolor. AB - The white rot fungus Trametes (Coriolus) versicolor can delignify and brighten unbleached hardwood kraft pulp within a few days, but softwood kraft pulps require longer treatment. To determine the contributions of higher residual lignin contents (kappa numbers) and structural differences in lignins to the recalcitrance of softwood kraft pulps to biobleaching, we tested softwood and hardwood pulps cooked to the same kappa numbers, 26 and 12. A low-lignin-content (overcooked) softwood pulp resisted delignification by T. versicolor, but a high lignin-content (lightly cooked) hardwood pulp was delignified at the same rate as a normal softwood pulp. Thus, the longer time taken by T. versicolor to brighten softwood kraft pulp than hardwood pulp results from the higher residual lignin content of the softwood pulp; possible differences in the structures of the residual lignins are important only when the lignin becomes highly condensed. Under the conditions used in this study, when an improved fungal inoculum was used, six different softwood pulps were all substantially brightened by T. versicolor. Softwood pulps whose lignin contents were decreased by extended modified continuous cooking or oxygen delignification to kappa numbers as low as 15 were delignified by T. versicolor at the same rate as normal softwood pulp. More intensive O(2) delignification, like overcooking, decreased the susceptibility of the residual lignin in the pulps to degradation by T. versicolor. PMID- 16349247 TI - Effect of Motility on Surface Colonization and Reproductive Success of Pseudomonas fluorescens in Dual-Dilution Continuous Culture and Batch Culture Systems. AB - The colonization of glass surfaces by motile and nonmotile strains of Pseudomonas fluorescens was evaluated by using dual-dilution continuous culture (DDCC), competitive and noncompetitive attachment assays, and continuous-flow slide culture. Both strains possessed identical growth rates whether in the attached or planktonic state. Results of attachment assays using radiolabeled bacteria indicated that both strains obeyed first-order (monolayer) adsorption kinetics in pure culture. However, the motile strain attached about four times more rapidly and achieved higher final cell densities on surfaces than did the nonmotile strain (2.03 x 10 versus 5.57 x 10 cells vial) whether evaluated alone or in cocultures containing motile and nonmotile P. fluorescens. These kinetics were attributed to the increased transport of motile cells from the bulk aqueous phase to the hydrodynamic boundary layer where bacterial attachment, growth, and recolonization could occur. First-order attachment kinetics were also observed for both strains by using continuous-flow slide culture assays analyzed by image analysis. The DDCC system contained both aqueous and particulate phases which could be diluted independently. DDCC results indicated that when cocultures containing motile and nonmotile P. fluorescens colonized solid particles, the motile strain replaced the nonmotile strain in the system over time. Increasing the aqueous-phase rates of dilution decreased the time required for extinction of the nonmotile strain while concurrently decreasing the overall carrying capacity of the DDCC system for both strains. These results confirmed that bacterial motility conveyed a selective advantage during surface colonization even in aqueous-phase systems not dominated by laminar flow. PMID- 16349248 TI - A Hydrophobic Mutant of Rhizobium etli Altered in Nodulation Competitiveness and Growth in the Rhizosphere. AB - We isolated and characterized CE3003, a Tn5-induced mutant with altered colony morphology derived from Rhizobium etli CE3. CE3003 produced domed colonies and was highly hydrophobic as indicated by its ability to partition into hexadecane, whereas its parent produced flat colonies and was hydrophilic. On bean plants, CE3003 induced nodules and reduced acetylene. CE3003 and CE3 grew at similar rates when they were grown separately or together in culture medium or inoculated singly onto bean seeds. However, when they were mixed at a 1:1 ratio and applied to seeds, CE3003 achieved significantly lower populations than CE3 in the rhizosphere. Five days after coinoculation of CE3 and CE3003, the population of the mutant was less than 10% of the population of CE3 in the bean rhizosphere. To determine the nodulation competitiveness of the mutant, it was coinoculated with CE3 at various ratios at planting, and the ratio of the nodules occupied by each strain was determined 21 days later. A 17,000-fold excess of CE3003 in mixed inocula was required to obtain equal nodule occupancy by the two strains. A genomic library of strain CE3 was mobilized into CE3003, and we identified a cosmid, pRA3003, that restored the parental colony morphology and hydrophilicity to the mutant. Restoration of the parental colony morphology was accompanied by recovery of the ability to grow competitively in the rhizosphere and to compete for nodulation of beans. The data show an association between cell surface hydrophobicity, nodulation competitiveness, and competitive growth in the rhizosphere in mutant CE3003. PMID- 16349249 TI - Thermophilic sulfate reduction in hydrothermal sediment of lake tanganyika, East Africa. AB - In environments with temperatures above 60 degrees C, thermophilic prokaryotes are the only metabolically active life-forms. By using the SO(4) tracer technique, we studied the activity of sulfate-reducing microorganisms (SRM) in hot sediment from a hydrothermal vent site in the northern part of freshwater Lake Tanganyika (East Africa). Incubation of slurry samples at 8 to 90 degrees C demonstrated meso- and thermophilic sulfate reduction with optimum temperatures of 34 to 45 degrees C and 56 to 65 degrees C, respectively, and with an upper temperature limit of 80 degrees C. Sulfate reduction was stimulated at all temperatures by the addition of short-chain fatty acids and benzoate or complex substrates (yeast extract and peptone). A time course experiment showed that linear thermophilic sulfate consumption occurred after a lag phase (12 h) and indicated the presence of a large population of SRM in the hydrothermal sediment. Thermophilic sulfate reduction had a pH optimum of about 7 and was completely inhibited at pH 8.8 to 9.2. SRM could be enriched from hydrothermal chimney and sediment samples at 60 and 75 degrees C. In lactate-grown enrichments, sulfide production occurred at up to 70 and 75 degrees C, with optima at 63 and 71 degrees C, respectively. Several sporulating thermophilic enrichments were morphologically similar to Desulfotomaculum spp. Dissimilatory sulfate reduction in the studied hydrothermal area of Lake Tanganyika apparently has an upper temperature limit of 80 degrees C. PMID- 16349250 TI - Effect of grain size on bacterial penetration, reproduction, and metabolic activity in porous glass bead chambers. AB - We determined the effects of grain size and nutritional conditions on the penetration rate and metabolic activity of Escherichia coli strains in anaerobic, nutrient-saturated chambers packed with different sizes of glass beads (diameters, 116 to 767 mum) under static conditions. The chambers had nearly equal porosities (38%) but different calculated pore sizes (range, 10 to 65 mum). Motile strains always penetrated faster than nonmotile strains, and nutrient conditions that resulted in faster growth rates (fermentative conditions versus nitrate-respiring conditions) resulted in faster penetration rates for both motile and nonmotile strains for all of the bead sizes tested. The penetration rate of nonmotile strains increased linearly when bead size was increased, while the penetration rate of motile strains became independent of the bead size when beads having diameters of 398 mum or greater were used. The rate of H(2) production and the final amount of H(2) produced decreased when bead size was decreased. However, the final protein concentrations were similar in chambers packed with 116-, 192-, and 281-mum beads and were only slightly higher in chambers packed with 398- and 767-mum beads. Our data indicated that conditions that favored faster growth rates also resulted in faster penetration times and that the lower penetration rates observed in chambers packed with small beads were due to restriction of bacterial activity in the small pores. The large increases in the final amount of hydrogen produced without corresponding increases in the final amount of protein made indicated that metabolism became uncoupled from cell mass biosynthesis as bead size increased, suggesting that pore size influenced the efficiency of substrate utilization. PMID- 16349251 TI - Diel Migrations of Microorganisms within a Benthic, Hypersaline Mat Community. AB - We studied the diel migrations of several species of microorganisms in a hypersaline, layered microbial mat. The migrations were quantified by repeated coring of the mat with glass capillary tubes. The resulting minicores were microscopically analyzed by using bright-field and epifluorescence (visible and infrared) microscopy to determine depths of coherent layers and were later dissected to determine direct microscopic counts of microorganisms. Microelectrode measurements of oxygen concentration, fiber optic microprobe measurements of light penetration within the mat, and incident irradiance measurements accompanied the minicore sampling. In addition, pigment content, photosynthesis and irradiance responses, the capacity for anoxygenic photosynthesis, and gliding speeds were determined for the migrating cyanobacteria. Heavily pigmented Oscillatoria sp. and Spirulina cf. subsalsa migrated downward into the mat during the early morning and remained deep until dusk, when upward migration occurred. The mean depth of the migration (not more than 0.4 to 0.5 mm) was directly correlated with the incident irradiance over the mat surface. We estimated that light intensity at the upper boundary of the migrating cyanobacteria was attenuated to such an extent that photoinhibition was effectively avoided but that intensities which saturated photosynthesis were maintained through most of the daylight hours. Light was a cue of paramount importance in triggering and modulating the migration of the cyanobacteria, even though the migrating phenomenon could not be explained solely in terms of a light response. We failed to detect diel migration patterns for other cyanobacterial species and filamentous anoxyphotobacteria. The sulfide-oxidizing bacterium Beggiatoa sp. migrated as a band that followed low oxygen concentrations within the mat during daylight hours. During the nighttime, part of this population migrated toward the mat surface, but a significant proportion remained deep. PMID- 16349252 TI - Role of neutral metabolites in microbial conversion of 3beta-acetoxy-19 hydroxycholest-5-ene into estrone. AB - Biotransformation of 3beta-acetoxy-19-hydroxycholest-5-ene (19-HCA, 6 g) by Moraxella sp. was studied. Estrone (712 mg) was the major metabolite formed. Minor metabolites identified were 5alpha-androst-1-en-19-ol-3,17-dione (33 mg), androst-4-en-19-ol-3,17-dione (58 mg), androst-4-en-9alpha,19-diol-3,17-dione (12 mg), and androstan-19-ol-3,17-dione (1 mg). Acidic metabolites were not formed. Time course experiments on the fermentation of 19-HCA indicated that androst-4-en 19-ol-3,17-dione was the major metabolite formed during the early stages of incubation. However, with continuing fermentation its level dropped, with a concomitant increase in estrone. Fermentation of 19-HCA in the presence of specific inhibitors or performing the fermentation for a shorter period (48 h) did not result in the formation of acidic metabolites. Resting-cell experiments carried out with 19-HCA (200 mg) in the presence of alpha,alpha'-bipyridyl led to the isolation of three additional metabolites, viz., cholestan-19-ol-3-one (2 mg), cholest-4-en-19-ol-3-one (10 mg), and cholest-5-en-3beta,19-diol (12 mg). Similar results were also obtained when n-propanol was used instead of alpha,alpha'-bipyridyl. Resting cells grown on 19-HCA readily converted both 5alpha-androst-1-en-19-ol-3,17-dione and androst-4-en-19-ol-3,17-dione into estrone. Partially purified 1,2-dehydrogenase from steroid-induced Moraxella cells transformed androst-4-en-19-ol-3,17-dione into estrone and formaldehyde in the presence of phenazine methosulfate, an artificial electron acceptor. These results suggest that the degradation of the hydrocarbon side chain of 19-HCA does not proceed via C(22) phenolic acid intermediates and complete removal of the C(17) side chain takes place prior to the aromatization of the A ring in estrone. The mode of degradation of the sterol side chain appears to be through the fission of the C(17)-C(20) bond. On the basis of these observations, a new pathway for the formation of estrone from 19-HCA in Moraxella sp. has been proposed. PMID- 16349253 TI - Reduction of Cr(VI) by a Consortium of Sulfate-Reducing Bacteria (SRB III). AB - A consortium of bacteria with tolerance to high concentrations of Cr(VI) (up to 2,500 ppm) and other toxic heavy metals has been obtained from metal-refinishing wastewaters in Chengdu, People's Republic of China. This consortium consists of a range of gram-positive and gram-negative rods and has the capacity to reduce Cr(VI) to Cr(III) as amorphous precipitates which are associated with the bacterial surfaces. An endospore-producing, gram-positive rod and a gram-negative rod accumulate the most metallic precipitates, and, over time, 80 to 95% of Cr can be removed from concentrations ranging from 50 to 2,000 ppm (0.96 to 38.45 mM). Kinetic studies revealed a first-order constant for Cr removal of 0.1518 h for an initial concentration of 1,000 ppm (19.3 mM), and the sorption isothermal data could be interpreted by the Freundlich relationship. The sorption was not entirely due to a passive interaction with reactive sites on the bacterial surfaces since gamma-irradiated, killed cells could not immobilize as much metal. When U or Zn was added with the Cr, it was also removed and could even increase the total amount of Cr immobilized. The consortium was tolerant to small amounts of oxygen in the headspace of tubes, but active growth of the bacteria was a requirement for Cr immobilization through Cr(VI) reduction, resulting in the lowering of E(h). Our data suggest that the reduction was via H(2)S. This consortium has been named SRB III, and it may be useful for the bioremediation of fluid metal-refining wastes. PMID- 16349254 TI - Limited Genetic Diversity in the Endophytic Sugarcane Bacterium Acetobacter diazotrophicus. AB - Acetobacter diazotrophicus isolates that originated from different sugarcane cultivars growing in diverse geographic regions of Mexico and Brazil were shown to have limited genetic diversity. Measurements of polymorphism in the electrophoretic mobilities of metabolic enzymes revealed that the mean genetic diversity per enzyme locus (among the four electrophoretic types distinguished) was 0.064. The results of the genetic analysis indicate that the genetic structure of A. diazotrophicus is clonal, with one largely predominant clone. Plasmids were present in 20 of 24 isolates, and the molecular sizes of the plasmids ranged from 2.0 to 170 kb. Two plasmids (a 20- to 24-kb plasmid detected in all 20 plasmid-containing isolates and a 170-kb plasmid observed in 14 isolates) were highly conserved among the isolates examined. Regardless of the presence of plasmids, all of the isolates shared a common pattern of nif structural gene organization on the chromosome. PMID- 16349255 TI - Compositional Changes in Compost during Composting and Growth of Agaricus bisporus. AB - Samples from conventional compost taken at various stages of composting and mushroom (Agaricus bisporus) growth were analyzed for changes in 80% ethanol and water extracts, monosaccharides in acid hydrolysates of polysaccharides, lignin concentration, and lignin structural features. Variable amounts of extraneous inorganic solids in the form of fine sandy particles were removed by sedimentation of the samples in a carbon tetrachloride-dibromomethane mixture. During composting, about two-thirds of the initial wall polysaccharides were consumed by compost microorganisms, and only 17% of the total polysaccharides were used during mushroom production. The relative lignin content of composts as measured by the acetyl bromide procedure increased, both during composting and mushroom growth, and the chemical structure of lignin was altered by condensation and oxidation reactions. PMID- 16349256 TI - Utilization of organic nitrogen sources by two phytoplankton species and a bacterial isolate in pure and mixed cultures. AB - Algal production of dissolved organic carbon and the regeneration of nutrients from dissolved organic carbon by bacteria are important aspects of nutrient cycling in the sea, especially when inorganic nitrogen is limiting. Dissolved free amino acids are a major carbon source for bacteria and can be used by phytoplankton as a nitrogen source. We examined the interactions between the phytoplankton species Emiliania huxleyi and Thalassiosira pseudonana and a bacterial isolate from the North Sea. The organisms were cultured with eight different amino acids and a protein as the only nitrogen sources, in pure and mixed cultures. Of the two algae, only E. huxleyi was able to grow on amino acids. The bacterium MD1 used all substrates supplied, except serine. During growth of MD1 in pure culture, ammonium accumulated in the medium. Contrary to the expectation, the percentage of ammonium regenerated from the amino acids taken up showed no correlation with the substrate C/N ratio. In mixed culture, the algae grew well in those cultures in which the bacteria grew well. The bacterial yields (cell number) were also higher in mixed culture than in pure culture. In the cultures of MD1 and T. pseudonana, the increase in bacterial yield (number of cells) over that of the pure culture was comparable to the bacterial yield in mixed culture on a mineral medium. This result suggests that T. pseudonana excreted a more-or-less-constant amount of carbon. The bacterial yields in mixed cultures with E. huxleyi showed a smaller and less consistent difference than those of the pure cultures of MD1. It is possible that the ability of E. huxleyi to use amino acids influenced the bacterial yield. The results suggest that interactions between algae and bacteria influence the regeneration of nitrogen from organic carbon and that this influence differs from one species to another. PMID- 16349257 TI - Effect of the Squid Host on the Abundance and Distribution of Symbiotic Vibrio fischeri in Nature. AB - Euprymna scolopes, a Hawaiian species of bioluminescent squid, harbors Vibrio fischeri as its specific light organ symbiont. The population of symbionts grew inside the adult light organ with an average doubling time of about 5 h, which produced an excess of cells that were expelled into the surrounding seawater on a diurnal basis at the beginning of each period of daylight. These symbionts, when expelled into the ambient seawater, maintain or slightly increase their numbers for at least 24 h. Hence, locations inhabited by their hosts periodically receive a daily input of symbiotic V. fischeri cells and, as a result, become significantly enriched with these bacteria. As estimated by hybridization with a species-specific luxA gene probe, the typical number of V. fischeri CFU, both in the water column and in the sediments of E. scolopes habitats, was as much as 24 to 30 times that in similar locations where squids were not observed. In addition, the number of symbiotic V. fischeri CFU in seawater samples that were collected along a transect through Kaneohe Bay, Hawaii, decreased as a function of the distance from a location inhabited by E. scolopes. These findings constitute evidence for the first recognized instance of the abundance and distribution of a marine bacterium being driven primarily by its symbiotic association with an animal host. PMID- 16349258 TI - Potential Role of Elicitins in the Interaction between Phytophthora Species and Tobacco. AB - The potential role of extracellular elicitor proteins (elicitins) from Phytophthora species as avirulence factors in the interaction between Phytophthora and tobacco was examined. A survey of 85 Phytophthora isolates representing 14 species indicated that production of elicitin is almost ubiquitous except for isolates of Phytophthora parasitica from tobacco. The production of elicitins by isolates of P. parasitica correlated without exception with low or no virulence on tobacco. Genetic analysis was conducted by using a cross between two isolates of P. parasitica, segregating for production of elicitin and virulence on tobacco. Virulence assays of the progeny on tobacco confirmed the correlation between production of elicitin and low virulence. PMID- 16349259 TI - Identification of chloroacetaldehyde dehydrogenase involved in 1,2-dichloroethane degradation. AB - The degradation of 1,2-dichloroethane and 2-chloroethanol by Xanthobacter autotrophicus GJ10 proceeds via chloroacetaldehyde, a reactive and potentially toxic intermediate. The organism produced at least three different aldehyde dehydrogenases, of which one is plasmid encoded. Two mutants of strain GJ10, designated GJ10M30 and GJ10M41, could no longer grow on 2-chloroethanol and were found to lack the NAD-dependent aldehyde dehydrogenase that is the predominant protein in wild-type cells growing on 2-chloroethanol. Mutant GJ10M30, selected on the basis of its resistance to 1,2-dibromoethane, also had lost haloalkane dehalogenase activity and Hg resistance, indicating plasmid loss. From a gene bank of strain GJ10, different clones that complemented one of these mutants were isolated. In both transconjugants, the aldehyde dehydrogenase that was absent in the mutants was overexpressed. The enzyme was purified and was a tetrameric protein of 55-kDa subunits. The substrate range was rather broad, with the highest activity measured for acetaldehyde. The K(m) value for chloroacetaldehyde was 160 muM, higher than those for other aldehydes tested. It is concluded that the ability of GJ10 to grow with 2-chloroethanol is due to the high expression level of an aldehyde dehydrogenase with a rather low activity for chloroacetaldehyde. PMID- 16349260 TI - Biochemical and Molecular Characterization of Obesumbacterium proteus, a Common Contaminant of Brewing Yeasts. AB - We have evaluated the effectiveness of API 20E, Biolog testing, plasmid profiling, ribotyping, and enteric repetitive intergenic consensus (ERIC)-PCR to characterize, classify, and differentiate nine bacterial isolates of the common brewery contaminant Obesumbacterium proteus. Of the five typing techniques, Biolog testing, plasmid profiling, and ERIC-PCR provided the most differentiation, and API 20E testing and ribotyping were relatively indiscriminate. The molecular biology approach of ERIC-PCR offered the ideal combination of speed, simplicity, and discrimination in this study. Overall, the results are supportive of the view that O. proteus can be subdivided into two biogroups, biogroup 1, which has considerable biochemical and genetic homology to Hafnia alvei, and biogroup 2, which is relatively heterogeneous. PMID- 16349261 TI - Evidence for Chlorine Migration during Oxidation of 2-Chlorobiphenyl by a Type II Methanotroph. AB - Axenic methanotrophic culture strain CSC1, which was derived from a mixed-culture aquifer sample, oxidized 2-chlorobiphenyl to a hydroxylated chlorobiphenyl intermediate identified as 2-hydroxy-3-chlorobiphenyl. Minor amounts of another hydroxylated chlorobiphenyl isomer, assumed to be the nonmigrated product 3 hydroxy-2-chlorobiphenyl, were found in axenic culture incubation extracts. The occurrence of chlorine migration during oxidation indicates that the metabolite is formed via a concerted oxidation mechanism involving an arene oxide, which rearranges spontaneously to the hydroxylated intermediate via an NIH shift mechanism. PMID- 16349262 TI - Evaluation of Fleroxacin Activity against Established Pseudomonas fluorescens Biofilms. AB - Scanning confocal laser microscopy (SCLM) and fluorescent molecular probes were used to evaluate the effect of the fluoroquinolone fleroxacin on the architecture of established Pseudomonas fluorescens biofilms. Control P. fluorescens biofilms were heterogeneous, consisting of cell aggregates extending from the attachment surface to maximum measured depths of approximately 90 mum (mean biofilm depth at 72 h, 42 +/- 28 mum) and penetrated by an array of channels. In contrast, fleroxacin-treated biofilms were less deep (mean biofilm depth at 72 h, 29 +/- 8 mum), varied little in depth over large areas, and consisted of a homogeneous distribution of cells. Fleroxacin also caused cells to elongate, with cells located near the biofilm-liquid interface lengthening significantly more than cells located at the attachment surface. By using SCLM, acridine orange, and image analysis it was found that approximately 59% of cells within fleroxacin treated biofilms emitted red fluorescence whereas >99% of cells from control biofilms emitted green fluorescence. The fleroxacin-treated cells which emitted red fluorescence were observed to be the population of cells which elongated. PMID- 16349263 TI - Tellurium and Selenium Resistance in Rhizobia and Its Potential Use for Direct Isolation of Rhizobium meliloti from Soil. AB - Forty-eight Rhizobium and Bradyrhizobium strains were screened for resistance to tellurite, selenite, and selenate. High levels of resistance to the metals were observed only in Rhizobium meliloti and Rhizobium fredii strains; the MICs were 2 to 8 mM for Te(IV), >200 mM for Se(VI), and 50 to 100 mM for Se(IV). Incorporation of Se and Te into growth media permitted us to directly isolate R. meliloti strains from soil. Mutant strains of rhizobia having decreased levels of Se and Te resistance were constructed by Tn5 mutagenesis and were found to have transposon insertions in DNA fragments of different sizes. Genomic DNAs from Te rhizobium strains failed to hybridize with Te determinants from plasmids RP4, pHH1508a, and pMER610. PMID- 16349264 TI - An Outbreak of Aeromonas hydrophila Infection in Turtles (Pseudemis scripta). AB - An outbreak of Aeromonas hydrophila infection with a high rate of mortality (95%) in turtles (Pseudemis scripta) in Italy is reported. Pure cultures of the pathogen were isolated from liver, lung, kidney, and heart specimens of the turtles. The A. hydrophila isolate was resistant to amoxicillin, ampicillin, cephalothin, and trimethoprim-sulfamethoxazole but was sensitive to a number of other antibiotics tested. The study indicates that pet turtles can act as reservoirs of this pathogen and may play an important role in the etiology of Aeromonas-associated human infections. PMID- 16349265 TI - Optimization of the Sistrom Culture Medium for Large-Scale Batch Cultivation of Rhodospirillum rubrum under Semiaerobic Conditions with Maximal Yield of Photosynthetic Membranes. AB - The defined medium A of W. R. Sistrom (W. R. Sistrom, J. Gen. Microbiol. 22:77 85, 1960) has been modified to allow the growth of Rhodospirillum rubrum in large scale batch cultures under dark, semiaerobic conditions. The simultaneous use of two substrates, NH(4)-succinate (46 mM) and fructose (0.3%), which are utilized in aerobic and fermentative metabolism, respectively, leads to very high cell densities with a maximal yield of photosynthetic membranes. PMID- 16349266 TI - An Enzyme-Linked Immunosorbent Assay-Based Isolation Procedure for Verotoxigenic Escherichia coli. AB - [This corrects the article on p. 4228 in vol. 59.]. PMID- 16349268 TI - Agglutination, adherence, and root colonization by fluorescent pseudomonads. AB - Two fractions of agglutination activity towards fluorescent pseudomonads were detected in root washes of potato, tomato, wheat, and bean. High-molecular-mass (>10 Da) components in crude root washes agglutinated only particular saprophytic, fluorescent Pseudomonas isolates. Ion-exchange treatment of the crude root washes resulted in preparations of lower-molecular-mass (10 to 10 Da) fractions which agglutinated almost all Pseudomonas isolates examined. Also, components able to suppress agglutination reactions of pseudomonads with the lower-molecular-mass root components were detected in crude root washes of all crops studied. Pseudomonas isolates were differentially agglutinated by both types of root components. The involvement of these two types of root components in short-term adherence and in colonization was studied in potato, tomato, and grass, using Pseudomonas isolates from these crops. Short-term adherence of isolates to roots was independent of their agglutination with either type of root components. With agglutination-negative mutants, the high-molecular-mass components seemed to be involved in adherence of Pseudomonas putida Corvallis to roots of all crops studied. Short-term adherence to roots of four Pseudomonas isolates could be influenced by addition of both crude and ion-exchange-treated root washes, depending on their agglutination phenotype with these root wash preparations. Potato root colonization by 10 different isolates from this crop, over a period of 7 days, was not correlated with their agglutination phenotype. Agg mutants of P. putida Corvallis were not impaired in root colonization. It is concluded that the root agglutinins studied can be involved in short-term adherence of pseudomonads to roots but do not play a decisive role in their root colonization. PMID- 16349267 TI - Genotypic Diversity among Bacillus cereus and Bacillus thuringiensis Strains. AB - Twenty-four strains of Bacillus cereus were analyzed by pulsed-field gel electrophoresis (PFGE) and compared with 12 Bacillus thuringiensis strains. In addition, the 36 strains were examined for variation in 15 chromosomal genes encoding enzymes (by multilocus enzyme electrophoresis [MEE]). The genome of each strain had a distinct NotI restriction enzyme digestion profile by PFGE, and the 36 strains could be assigned to 27 multilocus genotypes by MEE. However, neither PFGE nor MEE analysis could distinguish between the two species. Two of the B. cereus strains contained extrachromosomal DNA that hybridized to a cryIA insecticidal toxin probe, and seven strains contained DNA with homology to a Tn4430 transposon probe derived from B. thuringiensis. The results strongly indicate that B. cereus and B. thuringiensis should be regarded as one species. PMID- 16349269 TI - C Nuclear Magnetic Resonance Studies of Citrate and Glucose Cometabolism by Lactococcus lactis. AB - C nuclear magnetic resonance (C-NMR) was used to investigate the metabolism of citrate plus glucose and pyruvate plus glucose by nongrowing cells of Lactococcus lactis subsp. lactis 19B under anaerobic conditions. The metabolism of citrate plus glucose during growth was also monitored directly by in vivo NMR. Although pyruvate is a common intermediate metabolite in the metabolic pathways of both citrate and glucose, the origin of the carbon atoms in the fermentation products was determined by using selectively labeled substrates, e.g., [2,4-C]citrate, [3 C]pyruvate, and [2-C]glucose. The presence of an additional substrate caused a considerable stimulation in the rates of substrate utilization, and the pattern of end products was changed. Acetate plus acetoin and butanediol represented more than 80% (molar basis) of the end products of the metabolism of citrate (or pyruvate) alone, but when glucose was also added, 80% of the citrate (or pyruvate) was converted to lactate. This result can be explained by the activation of lactate dehydrogenase by fructose 1,6-bisphosphate, an intermediate in glucose metabolism. The effect of different concentrations of glucose on the metabolism of citrate by dilute cell suspensions was also probed by using analytical methods other than NMR. Pyruvate dehydrogenase (but not pyruvate formate-lyase) was active in the conversion of pyruvate to acetyl coenzyme A. alpha-Acetolactate was detected as an intermediate metabolite of citrate or pyruvate metabolism, and the labeling pattern of the end products agrees with the alpha-acetolactate pathway. It was demonstrated that the contribution of the acetyl coenzyme A pathway for the synthesis of diacetyl, should it exist, is lower than 10%. Evidence for the presence of internal carbon reserves in L. lactis is presented. PMID- 16349270 TI - Genetic structure and symbiotic characteristics of a bradyrhizobium population recovered from a pasture soil. AB - We examined the genetic structure and symbiotic characteristics of Bradyrhizobium isolates recovered from four legume species (Lupinus albus [white lupine], Lupinus angustifolius [blue lupine], Ornithopus compressus [yellow serradella], and Macroptilium atropurpureum [sirato]) grown in an Oregon soil. We established that multilocus enzyme electrophoresis (MLEE) can provide insights into the genetic relatedness among Bradyrhizobium strains by showing a positive correlation (r = >/=0.90) between the relatedness of Bradyrhizobium japonicum strains determined by MLEE at 13 enzyme loci and that determined by other workers using either DNA-DNA hybridization or DNA sequence divergence estimates. MLEE identified 17 electrophoretic types (ETs) among 95 Bradyrhizobium isolates recovered from the four hosts. Although the overall genetic diversity among the ETs (H = 0.69) is one of the largest measured to date in a local population of any soilborne bacterial species, there was no evidence of multilocus structure (linkage disequilibrium) within the population. The majority of the isolates (73%) were represented by two closely related ETs (2 and 3) which dominated the root nodules of white lupine, serradella, and siratro. In contrast, ET1 dominated nodules of blue lupine. Although representative isolates from all of the 17 ETs nodulated siratro, white lupine, blue lupine, and big trefoil (Lotus pedunculatus), they were either completely ineffective or poorly effective at fixing nitrogen on these hosts. Despite the widespread use of serradella as a surrogate host for lupine-nodulating bradyrhizobia, 7 of the 17 ETs did not nodulate this host, and the remaining 10 ETs were ineffective at fixing nitrogen. PMID- 16349271 TI - Increased Nitrogenase-Dependent H(2) Photoproduction by hup Mutants of Rhodospirillum rubrum. AB - Transposon Tn5 mutagenesis was used to isolate mutants of Rhodospirillum rubrum which lack uptake hydrogenase (Hup) activity. Three Tn5 insertions mapped at different positions within the same 13-kb EcoRI fragment (fragment E1). Hybridization experiments revealed homology to the structural hydrogenase genes hupSLM from Rhodobacter capsulatus and hupSL from Bradyrhizobium japonicum in a 3.8-kb EcoRI-ClaI subfragment of fragment E1. It is suggested that this region contains at least some of the structural genes encoding the nickel-dependent uptake hydrogenase of R. rubrum. At a distance of about 4.5 kb from the fragment homologous to hupSLM, a region with homology to a DNA fragment carrying hypDE and hoxXA from B. japonicum was identified. Stable insertion and deletion mutations were generated in vitro and introduced into R. rubrum by homogenotization. In comparison with the wild type, the resulting hup mutants showed increased nitrogenase-dependent H(2) photoproduction. However, a mutation in a structural hup gene did not result in maximum H(2) production rates, indicating that the capacity to recycle H(2) was not completely lost. Highest H(2) production rates were obtained with a mutant carrying an insertion in a nonstructural hup-specific sequence and with a deletion mutant affected in both structural and nonstructural hup genes. Thus, besides the known Hup activity, a second, previously unknown Hup activity seems to be involved in H(2) recycling. A single regulatory or accessory gene might be responsible for both enzymes. In contrast to the nickel-dependent uptake hydrogenase, the second Hup activity seems to be resistant to the metal chelator EDTA. PMID- 16349272 TI - Anaerobic Metabolism of Cyclohex-1-Ene-1-Carboxylate, a Proposed Intermediate of Benzoate Degradation, by Rhodopseudomonas palustris. AB - Anaerobic benzoate degradation by the phototrophic bacterium Rhodopseudomonas palustris has been proposed to proceed via aromatic ring reduction reactions leading to cyclohex-1-ene-1-carboxyl-coenzyme A (CoA) formation. The alicyclic product is then proposed to undergo three beta-oxidation-like modifications resulting in ring cleavage. Illuminated suspensions of benzoate-grown cells converted [7-C]cyclohex-1-ene-1-carboxylate to intermediates that comigrated with cyclohex-1-ene-1-carboxyl-CoA, 2-hydroxycyclohexanecar-boxyl-CoA, 2 ketocyclohexanecarboxyl-CoA, and pimelyl-CoA by thin-layer chromatography. This set of intermediates was also formed by cells grown anaerobically or aerobically on cyclohex-1-ene-1-carboxylate, indicating that benzoate-grown and cyclohex-1 ene-1-carboxylate-grown cells degrade this alicyclic acid by the same catabolic route. Four enzymatic activities proposed to be required for conversion of cyclohex-1-ene-1-carboxylate to pimelyl-CoA were detected at 3- to 10-fold-higher levels in benzoate-grown cells than in succinate-grown cells. These were cyclohex 1-ene-1-carboxylate-CoA ligase, cyclohex-1-ene-1-carboxyl-CoA hydratase, 2 hydroxycyclohexanecarboxyl-CoA dehydrogenase, and 2-ketocyclohexanecarboxyl-CoA hydrolase (ring cleaving). Pimelyl-CoA was identified in hydrolase reaction mixtures as the product of alicyclic ring cleavage. The results provide a first demonstration of an alicyclic ring cleavage activity. PMID- 16349273 TI - Accumulation of New Polypeptides in Ri T-DNA-Transformed Roots of Tomato (Lycopersicon esculentum) during the Development of Vesicular-Arbuscular Mycorrhizae. AB - Root-inducing transferred-DNA (Ri T-DNA)-transformed roots of tomato (Lycopersicon esculentum) were in vitro inoculated with surface-sterilized vesicular-arbuscular mycorrhizal leek root pieces. About 1 week after inoculation, the infection of the transformed root culture by the fungal endophyte was confirmed by photonic microscopy. Total proteins were extracted from the mycorrhizal roots and analyzed by two-dimensional polyacrylamide gel electrophoresis. Control gels were run with proteins extracted from noninoculated roots mixed with purified intraradical vesicles and extraradical hyphae. Comparison of the resulting patterns revealed the presence of two polypeptides with estimated apparent masses of 24 and 39 kDa that were detected only in infected roots. Polypeptides with similar migration parameters were not detected in roots challenged with spore extracts, suggesting that the accumulation of the polypeptides was directly linked to root colonization by the fungus rather than to induction by fungus-derived elicitors. PMID- 16349274 TI - Improved Method for Recovery of mRNA from Aquatic Samples and Its Application to Detection of mer Expression. AB - Previously described methods for extraction of mRNA from environmental samples may preclude detecting transcripts from genes that were present in low abundance in aquatic bacterial communities. By combining a boiling sodium dodecyl sulfate diethylpyrocarbonate lysis step with acid-guanidinium extraction, we improved recovery of target mRNA from both pure cultures and environmental samples. The most significant advantage of the new protocol is that it is easily adapted to yield high recovery of mRNA from 142-mm-diameter flat filters and high-capacity cartridge filters. The lysis and extraction procedures are more rapid than previously described methods, and many samples can be handled at once. RNA extracts have been shown to be free of contaminating DNA. The lysis procedure does not damage target mRNA sequences, and mRNA can be detected from fewer than 10 bacterial cells. We used the new method to examine transcripts of genes responsible for detoxification of mercurial compounds. Induction of merA (specifying mercuric reductase) transcripts in stationary-phase Pseudomonas aeruginosa containing Tn501 occurred within 60 s of HgCl(2) addition and was proportional to the amount of Hg(II) added. The new technique also allowed the detection of merA transcripts from the microbial community of a mercury contaminated pond (Reality Lake, Oak Ridge, Tenn.). Significant differences in merA transcript abundance were observed between different locations associated with the lake. The results indicate that the new method is simple and rapid and can be applied to the study of mer gene expression of aquatic communities in their natural habitats. PMID- 16349275 TI - Effect of Host Diet and Hindgut Microbial Composition on Cellulolytic Activity in the Hindgut of the American Cockroach, Periplaneta americana. AB - Cellulase activity measured as filter paper digesting activity (FPase) and carboxymethyl cellulase (CMCase) was demonstrated in hindgut extracts of the cockroach Periplaneta americana. The highest activities measured amounted to 0.89 and 0.12 U . ml for CMCase and FPase, respectively. The cellulolytic capacity of the hindgut population increased dramatically when protozoa were present, and the activities were found to vary depending on the feeding regimen. Cellulose-rich diets induced high protozoal numbers, resulting in a high cellulase activity. A close correlation was found between the number of Nyctotherus ovalis organisms, the major protozoans in the hindgut, and both FPase and CMCase activity. Since the numbers of this protozoan also correlated with the methane production of the insect, it appears that N. ovalis is responsible for the major part of cellulolytic and methanogenic activity found in the hindgut of P. americana. PMID- 16349276 TI - Substrate Preference in a Strain of Megasphaera elsdenii, a Ruminal Bacterium, and Its Implications in Propionate Production and Growth Competition. AB - The NIAH 1102 strain of Megasphaera elsdenii utilized lactate in preference to glucose when the two substrates were present. Even when lactate was supplied to cells fermenting glucose, the cells switched substrate utilization from glucose to lactate and did not utilize glucose until lactate decreased to a low concentration (1 to 2 mM). Since substrate utilization was shifted gradually without intermittence, typical diauxic growth was not seen. The cyclic AMP content did not rise markedly with the shift in substrate utilization, suggesting that this nucleotide is not involved in the regulation of the shift. It was unlikely that propionate was produced from glucose, which was explicable by the fact that lactate racemase activity dropped rapidly with the exhaustion of lactate and cells actively fermenting glucose did not possess this enzyme. A coculture experiment indicated that M. elsdenii NIAH 1102 is overcome by Streptococcus bovis JB1 in the competition for glucose, mainly because M. elsdenii NIAH 1102 is obliged to utilize lactate produced by S. bovis JB1; i.e., glucose utilization by M. elsdenii NIAH 1102 is suppressed by the coexistence of S. bovis JB1. PMID- 16349277 TI - Evolution of a Lytic Bacteriophage via DNA Acquisition from the Lactococcus lactis Chromosome. AB - We discovered a phage-host interaction in which the lytic phage ul36, in response to pressure exerted by an abortive phage resistance mechanism, acquired a large DNA fragment from the chromosome of Lactococcus lactis NCK203 to form a new phage, ul37. Phage ul37 was characterized at morphological, phenotypic, and genotypic levels and was found to be a member of the P335 species. Although it exhibits a high level of DNA homology with ul36, phage ul37 is resistant to the abortive mechanism and has a longer tail, a different base plate, and apparently a different origin of replication. The chromosomal DNA implicated in the formation of new phage ul37 was disrupted by site-specific integration in NCK203. This strategy prevented the appearance of ul37 during subsequent infections with ul36. PMID- 16349278 TI - Influence of Plumbing Materials on Biofilm Formation and Growth of Legionella pneumophila in Potable Water Systems. AB - A two-stage chemostat model of a plumbing system was developed, with tap water as the sole nutrient source. The model system was populated with a naturally occurring inoculum derived from an outbreak of Legionnaires' disease and containing Legionella pneumophila along with associated bacteria and protozoa. The model system was used to develop biofilms on the surfaces of a range of eight plumbing materials under controlled, reproducible conditions. The materials varied in their abilities to support biofilm development and the growth of L. pneumophila. Elastomeric surfaces had the most abundant biofilms supporting the highest numbers of L. pneumophila CFU; this was attributed to the leaching of nutrients for bacterial growth from the materials. No direct relationship existed between total biofouling and the numbers of L. pneumophila CFU. PMID- 16349279 TI - Assimilation of inorganic nitrogen by marine invertebrates and their chemoautotrophic and methanotrophic symbionts. AB - Symbioses between marine invertebrates and their chemoautotrophic and methanotrophic symbionts are now known to exist in a variety of habitats where reduced chemical species are present. The utilization of chemical energy and reliance on C(1) compounds by these symbioses are well documented. Much less is known about their metabolism of nitrogen. Earlier work has shown that the tissues of organisms in these associations are depleted of N compared with those of other marine organisms, indicating that local sources of nitrogen are assimilated and that novel mechanisms of nitrogen metabolism may be involved. Although these symbioses have access to rich sources of ammonium (NH(4) and NH(3)) and/or nitrate, several investigators have proposed that N(2) fixation may account for some of these isotope values. Here we report that [N]ammonium and, to a lesser degree, [N]nitrate are assimilated into organic compounds by Solemya reidi, a gutless clam containing S-oxidizing bacteria, and seep mussel Ia, an undescribed mytilid containing methanotrophic bacteria. In contrast, Riftia pachyptila, the giant hydrothermal vent tube worm symbiotic with S-oxidizing bacteria, assimilated nitrate but not exogenous ammonium. The rates of assimilation of these sources are sufficient to at least partially support C(1) compound metabolism. N(2) assimilation was not exhibited by the symbionts tested. PMID- 16349280 TI - Nitrogen Fixation and Hydrogen Metabolism in Relation to the Dissolved Oxygen Tension in Chemostat Cultures of the Wild Type and a Hydrogenase-Negative Mutant of Azorhizobium caulinodans. AB - Both the wild type and an isogenic hydrogenase-negative mutant of Azorhizobium caulinodans growing ex planta on N(2) as the N source were studied in succinate limited steady-state chemostat cultures under 0.2 to 3.0% dissolved O(2) tension. Production or consumption of O(2), H(2), and CO(2) was measured with an on-line connected mass spectrometer. In the range of 0.2 to 3.0%, growth of both the wild type and the mutant was equally dependent on the dissolved O(2) tension: the growth yield decreased, and the specific O(2) consumption and CO(2) production increased. A similar dependency on the dissolved O(2) tension was found for the mutant with 2.5% H(2) in the influent gas. The H(2)/N(2) ratio (moles of H(2) evolved per mole of N(2) consumed via nitrogenase) of the mutant, growing with or without 2.5% H(2), increased with increasing dissolved O(2) tensions. This increase in the H(2)/N(2) ratio was small but significant. The dependencies of the ATP/N(2) ratio (moles of ATP consumed per mole of N(2) fixed) and the ATP/2e ratio [moles of ATP consumed per mole of electron pairs transferred from NAD(P)H to nitrogenase] on the dissolved O(2) tension were estimated. These dependencies were interpreted in terms of the physiological concepts of respiratory protection and autoprotection. PMID- 16349281 TI - Colloidal and Dissolved Organic Matter Excreted by a Mixotrophic Flagellate during Bacterivory and Autotrophy. AB - Excretion of dissolved and colloidal organic carbon by a mixotrophic flagellate, the chrysophyte Poterioochromonas malhamensis, was studied. Flagellates were incubated either with C-labeled bacteria or with inorganic C, in order to compare organic exudates originating from primary production with exudates originating from ingested bacteria. Colloids of >0.02 mum constituted a larger fraction of the exudates originating from ingested bacteria, compared with exudates derived from primary production. Flagellate feeding on bacteria specifically labeled in different cell components was compared. Cell wall components gave rise to less colloidal organic carbon than did other cell constituents. To investigate the degradability of flagellate C-exudates, they were added to lake water and mineralization to CO(2) was monitored. Bacterially derived exudates were more recalcitrant than exudates originating from photosynthesis. The results support the hypothesis that bacterial utilization of labile organic compounds, followed by flagellate bacterivory and exudation, results in a transformation of labile organic matter into more recalcitrant forms. PMID- 16349282 TI - Transformations of Aromatic Compounds by Nitrosomonas europaea. AB - Benzene and a variety of substituted benzenes inhibited ammonia oxidation by intact cells of Nitrosomonas europaea. In most cases, the inhibition was accompanied by transformation of the aromatic compound to a more oxidized product or products. All products detected were aromatic, and substituents were often oxidized but were not separated from the benzene ring. Most transformations were enhanced by (NH(4))(2)SO(4) (12.5 mM) and were prevented by C(2)H(2), a mechanism based inactivator of ammonia monooxygenase (AMO). AMO catalyzed alkyl substituent hydroxylations, styrene epoxidation, ethylbenzene desaturation to styrene, and aniline oxidation to nitrobenzene (and unidentified products). Alkyl substituents were preferred oxidation sites, but the ring was also oxidized to produce phenolic compounds from benzene, ethylbenzene, halobenzenes, phenol, and nitrobenzene. No carboxylic acids were identified. Ethylbenzene was oxidized via styrene to two products common also to oxidation of styrene; production of styrene is suggestive of an electron transfer mechanism for AMO. Iodobenzene and 1,2-dichlorobenzene were oxidized slowly to halophenols; 1,4-dichlorobenzene was not transformed. No 2-halophenols were detected as products. Several hydroxymethyl (-CH(2)OH)-substituted aromatics and p-cresol were oxidized by C(2)H(2)-treated cells to the corresponding aldehydes, benzaldehyde was reduced to benzyl alcohol, and o-cresol and 2,5-dimethylphenol were not depleted. PMID- 16349283 TI - A biological sensor for iron available to bacteria in their habitats on plant surfaces. AB - A sensor responsive to iron was constructed by fusing a promoterless ice nucleation activity gene (inaZ) to an iron-regulated promoter of a genomic region involved in pyoverdine (fluorescent siderophore) (pvd) production in Pseudomonas syringae. Cells of Pseudomonas fluorescens and P. syringae that contained the pvd inaZ fusion expressed iron-responsive ice nucleation activity in the bean rhizosphere and phyllosphere, respectively, and in culture. Addition of Fe(III) to leaves or soil reduced the apparent transcription of the pvd-inaZ reporter gene, as shown by a reduction in the number of ice nuclei produced, indicating that Fe(III) was primarily responsible for mediating transcription of the pvd inaZ gene even in natural environments. A Pseudomonas sp. strain having an intact iceC gene, which conferred Fe-insensitive expression of ice nucleation activity, was included in all studies to account for small strain- or environment-dependent differences in the ability of bacterial cells to produce ice nuclei. Thus, a comparison of the ice nucleation activity conferred by pvd-inaZ with the activity conferred by iceC revealed the bioavailability of iron in culture or natural habitats. The relative ice nucleation activities expressed by strains containing iceC or pvd-inaZ indicated that, while not abundant, Fe(III) is not present at extremely low concentrations at all microsites colonized by bacteria on plant surfaces. Biological sensors that are constructed by fusing inaZ to chemically responsive promoters provide a novel way to characterize chemical constituents of microbial habitats. PMID- 16349284 TI - Autotrophic, hydrogen-oxidizing, denitrifying bacteria in groundwater, potential agents for bioremediation of nitrate contamination. AB - Addition of hydrogen or formate significantly enhanced the rate of consumption of nitrate in slurried core samples obtained from an active zone of denitrification in a nitrate-contaminated sand and gravel aquifer (Cape Cod, Mass.). Hydrogen uptake by the core material was immediate and rapid, with an apparent K(m) of 0.45 to 0.60 muM and a V(max) of 18.7 nmol cm h at 30 degrees C. Nine strains of hydrogen-oxidizing denitrifying bacteria were subsequently isolated from the aquifer. Eight of the strains grew autotrophically on hydrogen with either oxygen or nitrate as the electron acceptor. One strain grew mixotrophically. All of the isolates were capable of heterotrophic growth, but none were similar to Paracoccus denitrificans, a well-characterized hydrogen-oxidizing denitrifier. The kinetics for hydrogen uptake during denitrification were determined for each isolate with substrate depletion progress curves; the K(m)s ranged from 0.30 to 3.32 muM, with V(max)s of 1.85 to 13.29 fmol cell h. Because these organisms appear to be common constituents of the in situ population of the aquifer, produce innocuous end products, and could be manipulated to sequentially consume oxygen and then nitrate when both were present, these results suggest that these organisms may have significant potential for in situ bioremediation of nitrate contamination in groundwater. PMID- 16349285 TI - Lipid Peroxidation by the Manganese Peroxidase of Phanerochaete chrysosporium Is the Basis for Phenanthrene Oxidation by the Intact Fungus. AB - The manganese peroxidase (MnP) of Phanerochaete chrysosporium supported Mn(II) dependent, H(2)O(2)-independent lipid peroxidation, as shown by two findings: linolenic acid was peroxidized to give products that reacted with thiobarbituric acid, and linoleic acid was peroxidized to give hexanal. MnP also supported the slow oxidation of phenanthrene to 2,2'-diphenic acid in a reaction that required Mn(II), oxygen, and unsaturated lipids. Phenanthrene oxidation to diphenic acid by intact cultures of P. chrysosporium occurred to the same extent that oxidation in vitro did and was stimulated by Mn. These results support a role for MnP mediated lipid peroxidation in phenanthrene oxidation by P. chrysosporium. PMID- 16349286 TI - Mode of Action of Nisin Z against Listeria monocytogenes Scott A Grown at High and Low Temperatures. AB - Nisin Z, a natural nisin variant, was recently isolated from Lactococcus lactis subspecies lactis NIZO 22186. The gene for this lantibiotic, designated nisZ, has been cloned, and its nucleotide sequence was found to be identical to that of the precursor nisin gene with the exception of a single mutation resulting in the substitution of Asn-27 for His-27 in the mature polypeptide (J. W. M. Mulders, I. J. Boerrigter, H. S. Rollema, R. J. Siezen, and W. M. de Vos, Eur. J. Biochem. 201:581-584, 1991). A K electrode was used to investigate the effect of various environmental parameters on the action of nisin Z against Listeria monocytogenes. Addition of nisin Z resulted in immediate loss of cell K, depolarization of the cytoplasmic membrane, inhibition of respiratory activity, and hydrolysis and partial efflux of cellular ATP. The action of nisin Z was optimal at pH 6.0 and was significantly reduced by di- and trivalent cations. The lanthanide gadolinium (Gd) was an efficient inhibitor and prevented nisin Z activity completely at a concentration of 0.2 mM. Nisin Z-induced loss of cell K was reduced at low temperatures, presumably as a result of the increased ordering of the lipid hydrocarbon chains in the cytoplasmic membrane. In cells grown at 30 degrees C, the action of nisin Z was prevented below 7 degrees C, whereas in cells grown at 4 degrees C nisin Z was able to induce K leakage at this low temperature. PMID- 16349287 TI - Enzyme Immunoassay Detection of Nitrosomonas europaea. AB - An exploratory effort to selectively detect the presence of a nitrifying bacterium, Nitrosomonas europaea, successfully demonstrated the fundamental utility of an enzyme-based immunoassay protocol. The applied polyclonal antibody test seemingly offered a marked improvement over the available analytical options, including plating, activity, and fluorescence immunoassay techniques. Following an initial purification step to enhance overall specificity, this procedure had an apparent lower limit of detection of approximately 5 x 10 cells per ml. Tests conducted with activated sludge samples exhibited a distinct difference between nitrifying and nonnitrifying mixed liquors, although the highest Nitrosomonas levels observed (i.e., at 1 to 2% of the overall viable cell density) were relatively close to the latter detection boundary. PMID- 16349288 TI - Effects of Temperature, pH, and NaCl on Growth and Pectinolytic Activity of Pseudomonas marginalis. AB - The interaction of temperature (4, 10, 18, and 30 degrees C), pH (6, 7, and 8), and NaCl (0, 2.5, and 5%) and their effects on specific growth rate, lag phase, and pectinolytic enzymes of Pseudomonas marginalis were evaluated. Response surface methodology was adapted to describe the response of growth parameters to environmental changes. To obtain good conditions of storage, the combined action of salt and temperature is necessary. At 4 degrees C with an NaCl concentration of 5% and a pH of 7, the lag time was 8 days and no growth was observed at 4 degrees C with 5% NaCl and a pH of 6. In the absence of salt, P. marginalis could grow regardless of temperature and pH. Pectate lyase and pectin lyase were produced by P. marginalis, while pectin methyl esterase activity was not observed in our culture conditions. The enzyme production depended on temperature, pH, and salt concentration but also on the age of the culture. Pectinolytic enzymes were abundantly excreted during the stationary phase, and even at 4 degrees C, after 2 weeks of storage, enzyme activities in supernatant culture were sufficient to damage vegetables. Both bacterial growth and enzymatic production have to be taken into account in order to estimate correctly the shelf life of vegetables. PMID- 16349289 TI - Pyrrolnitrin Production by Biological Control Agent Pseudomonas cepacia B37w in Culture and in Colonized Wounds of Potatoes. AB - Bacterial strain B37w (= NRRL B-14858), an isolate noteworthy because it inhibits the growth of the bioherbicide fungus Colletotrichum truncatum, was selected for further studies of bacterial antifungal properties. This isolate was identified as a Pseudomonas cepacia strain by performing carbohydrate utilization and fatty acid profile analyses, as well as other biochemical and physiological tests. Petri plate assays revealed that strain B37w exhibited antifungal activity against the potato dry rot fungus Fusarium sambucinum. Using bioautography, we correlated antifungal activity with production of a specific compound. Isolation from strain B37w and identification of the antifungal antibiotic pyrrolnitrin are described. A whole-potato assay revealed B37w's ability to colonize potato wounds. Wounded potatoes were inoculated with B37w, and pyrrolnitrin was detected in these potatoes by thin-layer chromatography-bioautography at a concentration on the order of nanograms per wound. We performed an assay in which we examined efficacy against F. sambucinum-incited potato dry rot and found that B37w inhibited disease development. This is the first report of P. cepacia or pyrrolnitrin activity against the economically important potato pathogen F. sambucinum. PMID- 16349290 TI - Sunlight inactivation of enterococci and fecal coliforms in sewage effluent diluted in seawater. AB - Inactivation (loss of culturability) by sunlight of enterococci and fecal coliforms within sewage effluent diluted in seawater was investigated in field experiments. In most experiments, 500-ml flasks of pure silica were used to confine activated sludge effluent diluted to 2% (vol/vol) in seawater. Inactivation of bacteria in these flasks (diameter, 0.1 m) was faster than in either open chambers (depth, 0.25 m) or patches of dyed effluent (depth of order, 1 m), probably because of the longer light paths in the latter two types of experiment, which caused greater attenuation of sunlight. Inactivation of 90% of enterococci generally required 2.3 times the insolation required for 90% inactivation of fecal coliforms, because of both the presence of larger initial shoulders on survival curves and a lower final inactivation rate. Two parameters are required to model inactivation of enterococci, a shoulder constant as well as a rate coefficient. The depth dependence of inactivation rate for both fecal indicators matched the attenuation profile of UV-A radiation at about 360 nm. Inactivation by UV-B radiation (290 to 320 nm), which penetrates much less into seawater, is of minor importance compared with the UV-A and visible radiation in sunlight, contrary to expectations in consideration of published action spectra for bacterial inactivation. PMID- 16349291 TI - Use of Lactobacillus plantarum LPCO10, a Bacteriocin Producer, as a Starter Culture in Spanish-Style Green Olive Fermentations. AB - Bacteriocin-producing Lactobacillus plantarum LPCO10 and its non-bacteriocin producing, bacteriocinimmune derivative, L. plantarum 55-1, were evaluated separately for growth and persistence in natural Spanish-style green olive fermentations. Both strains were genetically marked and selectively enumerated using antibiotic-containing media. Plasmid profile and bacteriocin production (bac) were used as additional markers. When olive brines were inoculated at 10 CFU/ml, the parent strain, LPCO10, proliferated to dominate the epiphytic microflora, sharing high population levels with other spontaneously occurring lactobacilli and persisting throughout the fermentation (12 weeks). In contrast, the derivative strain could not be isolated after 7 weeks. Stability of both plasmid profile and bac (LPCO10 strain) or bac (55-1 strain) phenotype was shown by L. plantarum LPCO10 and L. plantarum 55-1 isolated throughout the fermentation. Bacteriocin activity could be found in the L. plantarum LPCO10 inoculated brines only after ammonium sulfate precipitation and concentration (20 times) of the final brine. Spontaneously occurring lactobacilli and lactic coccus populations, which were isolated from each of the fermenting brines studied during this investigation, were shown to be sensitive to the bacteriocins produced by L. plantarum LPCO10 when tested by the drop diffusion test. The declines in both pH and glucose levels throughout the fermentative process were similar in L. plantarum LPCO10- and in L. plantarum 55-1-inoculated brines and were comparable to the declines in the uninoculated brines. However, the final concentration of lactic acid in L. plantarum LPCO10-inoculated brines was higher than in the L. plantarum 55-1-inoculated brines and uninoculated brines. These results indicated that L. plantarum LPCO10 may be useful as a starter culture to control the lactic acid fermentation of Spanish-style green olives. PMID- 16349292 TI - The DeLey-Doudoroff Pathway of Galactose Metabolism in Azotobacter vinelandii. AB - Azotobacter vinelandii cell extracts reduced NAD and oxidized d-galactose to galactonate that subsequently was converted to 2-keto-3-deoxy-galactonate. Further metabolism of 2-keto-3-deoxy-galactonate required the presence of ATP and resulted in the formation of pyruvate and glyceraldehyde 3-P. Radiorespirometry indicated a preferential release of CO(2) at the first carbon position of the d galactose molecule. This suggested that Azotobacter vinelandii metabolizes d galactose via the DeLey-Doudoroff pathway. The first enzyme of this pathway, d galactose dehydrogenase, was partially characterized. It has a molecular weight of about 74,000 Da and an isoelectric point of 6.15. The pH optimum of the galactose dehydrogenase was about 9. The apparent K(m)s for NAD and d-galactose were 0.125 and 0.56 mM, respectively. Besides d-galactose, the active fraction of this galactose dehydrogenase also oxidized l-arabinose effectively. The electron acceptor for d-galactose or l-arabinose oxidation, NAD, could not be replaced by NADP. These substrate specificities were different from those reported in Pseudomonas saccharophila, Pseudomonas fluorescens, and Rhizobium meliloti. PMID- 16349293 TI - Characterization of Monoclonal Antibodies Specific for Erwinia carotovora subsp. atroseptica and Comparison of Serological Methods for Its Sensitive Detection on Potato Tubers. AB - Seven monoclonal antibodies (MAbs) to Erwinia carotovora subsp. atroseptica have been produced. One, called 4G4, reacted with high specificity for serogroup I of E. carotovora subsp. atroseptica, the most common serogroup on potato tubers in different serological assays. Eighty-six strains belonging to different E. carotovora subsp. atroseptica serogroups were assayed. Some strains of serogroup XXII also reacted positively. No cross-reactions were observed against other species of plant pathogenic bacteria or 162 saprophytic bacteria from potato tubers. Only one strain of E. chrysanthemi from potato cross-reacted. A comparison of several serological techniques to detect E. carotovora subsp. atroseptica on potato tubers was performed with MAb 4G4 or polyclonal antibodies. The organism was extracted directly from potato peels of artificially inoculated tubers by soaking or selective enrichment under anaerobiosis in a medium with polypectate. MAb 4G4 was able to detect specifically 240 E. carotovora subsp. atroseptica cells per ml by indirect immunofluorescence and immunofluorescence colony staining and after soaking by ELISA-DAS (double-antibody sandwich enzyme linked immunosorbent assay) after enrichment. The same amount of cells was detected by using immunolectrotransfer with polyclonal antibodies, and E. carotovora subsp. atroseptica and subsp. carotovora were distinguished by the latter technique. ELISA-DAS using MAb 4G4 with an enrichment step also efficiently detected E. carotovora subsp. atroseptica in naturally infected tubers and plants. PMID- 16349294 TI - Uncoupling of bacterioplankton and phytoplankton production in fresh waters is affected by inorganic nutrient limitation. AB - Pelagic bacterial production is often positively correlated, or coupled, with primary production through utilization of autotrophically produced dissolved organic carbon. Recent studies indicate that inorganic N or P can directly limit both bacterial and phytoplanktonic growth. Our mesocosm experiments, with whole communities from mesotrophic Calder Lake, test whether this apparent bacterial algal coupling may be the result of independent responses to limiting inorganic nutrients. In systems without N additions, numbers of bacteria but not phytoplankton increased 2- to 2.5-fold in response to P fertilization (0 to 2.0 mumol of P per liter); this resulted in uncoupled production patterns. In systems supplemented with 10 mumol of NH(4)NO(3) per liter, P addition resulted in up to threefold increases in bacteria and two- to fivefold increases in total phytoplankton biomass (close coupling). P limitation of pelagic bacteria occurred independently of phytoplankton dynamics, and regressions between bacterial abundance and phytoplankton chlorophyll a were nonsignificant in all systems without added N. We describe a useful and simple coupling index which predicts that shifts in phytoplankton and bacterioplankton growth will be unrelated (Delta bacteria/Delta phytoplankton --> either + infinity or - infinity) in systems with inorganic N/P (molar) ratios of < approximately 40. In systems with higher N/P ratios (>40), the coupling index will approach 1.0 and close coupling between bacteria and phytoplankton is predicted to occur. PMID- 16349295 TI - Estimation of nitrification and denitrification from microprofiles of oxygen and nitrate in model sediment systems. AB - The coupling between nitrification and denitrification and the regulation of these processes by oxygen were studied in freshwater sediment microcosms with O(2) and NO(3) microsensors. Depth profiles of nitrification (indicated as NO(3) production), denitrification (indicated as NO(3) consumption), and O(2) consumption activities within the sediment were calculated from the measured concentration profiles. From the concentration profiles, it was furthermore possible to distinguish between the rate of denitrification based on the diffusional supply of NO(3) from the overlying water and the rate based on NO(3) supplied by benthic nitrification (D(w) and D(n), respectively). An increase in O(2) concentration caused a deeper O(2) penetration while a decrease in D(w) and an increase in D(n) were observed. The relative importance for total denitrification of NO(3) produced by nitrification thus increased compared with NO(3) supplied from the water phase. The decrease in D(w) at high oxygen was due to an increase in diffusion path for NO(3) from the overlying water to the denitrifying layers in the anoxic sediment. At high O(2) concentrations, nitrifying activity was restricted to the lower part of the oxic zone where there was a continuous diffusional supply of NH(4) from deeper mineralization processes, and the long diffusion path from the nitrification zone to the overlying water compared with the path to the denitrifying layers led to a stimulation in D(n). PMID- 16349296 TI - Pectinolytic enzymes from actinomycetes for the degumming of ramie bast fibers. AB - Actinomycetes isolated from 10 different soil and compost samples were screened for production of pectinolytic enzyme activities when grown on pectin-containing solid and liquid media. Pectinolytic enzymes, detected by using plate diffusion tests with a medium containing ramie (Boehmeria nivea) plant material as the sole carbon source, were mainly pectate lyases, but low activities of pectinesterases were also observed. Polygalacturonases and polymethylgalacturonases were not produced. Multiple forms of pectate lyases were detected in the culture supernatants of some of the strains by using the zymogram technique of isoelectric focusing gels. Xylanolytic and cellulolytic activities were always found to be associated with pectinolytic activities. None of the pectinolytic enzymes were produced in a medium with glucose as the sole carbon source. Treatment of ramie bast fibers with crude enzyme preparations from a selection of strains showed a good correlation between the pectate lyase activity applied and the degumming effect, resulting in good separation of the bast fibers. PMID- 16349297 TI - Root-to-Root Travel of the Beneficial Bacterium Azospirillum brasilense. AB - The root-to-root travel of the beneficial bacterium Azospirillum brasilense on wheat and soybean roots in agar, sand, and light-textured soil was monitored. We used a motile wild-type (Mot) strain and a motility-deficient (Mot) strain which was derived from the wild-type strain. The colonization levels of inoculated roots were similar for the two strains. Mot cells moved from inoculated roots (either natural or artificial roots in agar, sand, or light-textured soil) to noninoculated roots, where they formed a band-type colonization composed of bacterial aggregates encircling a limited part of the root, regardless of the plant species. The Mot strain did not move toward noninoculated roots of either plant species and usually stayed at the inoculation site and root tips. The effect of attractants and repellents was the primary factor governing the motility of Mot cells in the presence of adequate water. We propose that interroot travel of A. brasilense is an essential preliminary step in the root bacterium recognition mechanism. Bacterial motility might have a general role in getting Azospirillum cells to the site where firmer attachment favors colonization of the root system. Azospirillum travel toward plants is a nonspecific active process which is not directly dependent on nutrient deficiency but is a consequence of a nonspecific bacterial chemotaxis, influenced by the balance between attractants and possibly repellents leaked by the root. PMID- 16349298 TI - Siderophore-Mediated Iron Sequestering by Shewanella putrefaciens. AB - The iron-sequestering abilities of 51 strains of Shewanella putrefaciens isolated from different sources (fish, water, and warm-blooded animals) were assessed. Thirty strains (60%) produced siderophores in heat-sterilized fish juice as determined by the chrome-azurol-S assay. All cultures were negative for the catechol-type siderophore, whereas 24 of the 30 siderophore-producing strains tested positive in the Csaky test, indicating the production of siderophores of the hydroxamate type. Siderophore-producing S. putrefaciens could to some degree cross-feed on the siderophores of other S. putrefaciens strains and on compounds produced by an Aeromonas salmonicida strain under iron-limited conditions. The siderophores of S. putrefaciens were not sufficiently strong to inhibit growth of other bacteria under iron-restricted conditions. However, siderophore-producing Pseudomonas bacteria were always inhibitory to S. putrefaciens under iron-limited conditions. Growth of siderophore-producing strains under iron-limited conditions induced the formation of one major new outer membrane protein of approximately 72 kDa. Two outer membrane proteins of approximately 53 and 23 kDa were not seen when iron was restricted. PMID- 16349299 TI - Occurrence of fumonisins b(1) and b(2) in corn-based products from the spanish market. AB - The natural occurrence of fumonisins B(1) and B(2), the incidence of Fusarium organisms, and the capacity of Fusarium isolates to produce fumonisins were investigated with 50 corn-based samples from Spain destined for human consumption. Eight samples (16%) were found to be contaminated with fumonisins. The levels of contamination were very low, with a mean of 80 ng/g. PMID- 16349300 TI - Buffering Capacity of Pigmented and Nonpigmented Strains of Serratia marcescens. AB - The pigmented strain Serratia marcescens ATCC 274 had a higher buffering capacity and a higher membrane H conductance than S. marcescens GP, a spontaneous nonpigmented mutant of ATCC 274. The data suggest that mutations which apparently affect only the synthesis of a secondary metabolite can modify buffering capacity and passive H conductance. PMID- 16349301 TI - Influence of Serum and Glucose Additives on Survival of Actinobacillus pleuropneumoniae Aerosolized from the Freeze-Dried State. AB - Serum and/or glucose added to Actinobacillus pleuropneumoniae suspensions before freeze-drying significantly increased survival rates of bacteria in aerosols. Aerosols with predictable numbers of viable bacteria can be made as required in an aerosol infection model. Sucrose supplementation of impinger fluids increased recovery of viable A. pleuropneumoniae. PMID- 16349302 TI - Efficacy of Chemical Disinfectants against Turbot Aquareovirus. AB - The susceptibility of turbot aquareovirus to five chemical agents was examined. Treatment with 5 mg of malachite green per liter or 500 mg of iodine per liter resulted in a 90% reduction in virus titer within 1 h. Complete inactivation within 5 min was obtained with 2% formalin, 42.5% isopropanol, or 15 mg of free available chlorine per liter. Lower concentrations of chlorine were ineffective. PMID- 16349303 TI - Microbial transformation of nitroaromatics in surface soils and aquifer materials. AB - Microorganisms indigenous to surface soils and aquifer materials collected at a munitions-contaminated site transformed 2,4,6-trinitrotoluene (TNT), 2,4 dinitrotoluene (2,4-DNT), and 2,6-dinitrotoluene (2,6-DNT) to amino-nitro intermediates within 20 to 70 days. Carbon mineralization studies with both unlabeled (TNT, 2,4-DNT, and 2,6-DNT) and radiolabeled ([C]TNT) substrates indicated that a significant fraction of these source compounds was degraded to CO(2). PMID- 16349304 TI - Helicobacter pylori comb. nov. Exhibits Facultative Acidophilism and Obligate Microaerophilism. AB - Modified brucella broth medium was used to study the growth of Helicobacter pylori at varied pHs and partial pressures of oxygen and to determine the effect of urea on culture pH. Our findings suggested that the pHs of the media remained stable with or without urea and that H. pylori showed facultative acidophilism and obligate microaerophilism. PMID- 16349305 TI - Isolation and Identification of Environmental Mycobacteria in the Mycobacterium bovis BCG Trial Area of South India. AB - The isolation profiles of environmental mycobacteria present in soil, water, and dust samples, and sputum samples of persons with symptoms of chest infection in the South Indian Mycobacterium bovis BCG (bacillus Calmette-Guerin) trial area were compared. Isolates belonging to the Mycobacterium avium-intracellulare scrofulaceum complex were predominant in water, dust, and sputum samples and Mycobacterium fortuitum-complex organisms were predominant in soil samples irrespective of the season of the year. PMID- 16349306 TI - Efficacy of Selected Respiratory Protective Equipment Challenged with Bacillus subtilis subsp. niger. AB - The efficacy of powered air-purifying respirators, surgical masks, dust/mist respirators, and high-efficiency respirators were tested with a biological aerosol under simulated breathing conditions. Protective ability ranged from 67 to 99.95%. The majority of penetration in negative-pressure respirators occurred at the face-mask interface rather than through the filter material. PMID- 16349307 TI - Photometabolism of Heterocyclic Aromatic Compounds by Rhodopseudomonas palustris OU 11. AB - Rhodopseudomonas palustris OU 11 (ATCC 51186; DSM 7375) isolated from a pond of chemical industry effluent could anaerobically photometabolize heterocyclic aromatic compounds belonging to the pyridine and pyrazine groups only after a period of adaptation on pyrazinoic acid of 5 to 6 weeks. Growth on heterocyclic compounds was light dependent. The effects of various concentrations of heterocyclic compounds on growth suggest that higher concentrations of these compounds inhibit growth and are toxic. PMID- 16349308 TI - Reductive Dechlorination of Trichloroethylene and Tetrachloroethylene under Aerobic Conditions in a Sediment Column. AB - Biodegradation of trichloroethylene and tetrachloroethylene under aerobic conditions was studied in a sediment column. Cumulative mass balances indicated 87 and 90% removal for trichloroethylene and tetrachloroethylene, respectively. These studies suggest the potential for simultaneous aerobic and anaerobic biotransformation processes under bulk aerobic conditions. PMID- 16349309 TI - Effect of Biocides on MS2 and K Coliphages. AB - Several biocides commonly used in disinfection processes as antibacterial and antifungal agents were tested for activity against MS2 and K coliphages. MS2 was resistant to most biocides; only glutaraldehyde (0.5%) and peracetic acid (1%) achieved a 4-log(10) titer reduction in 20 min. In contrast, K phage was sensitive to most biocides, being resistant only to phenol (2%) and chlorhexidine (1%). PMID- 16349310 TI - Use of the Escherichia coli beta-Glucuronidase (gusA) Gene as a Reporter Gene for Analyzing Promoters in Lactic Acid Bacteria. AB - [This corrects the article on p. 588 in vol. 60.]. PMID- 16349311 TI - Anaerobic degradation of normal- and branched-chain Fatty acids with four or more carbons to methane by a syntrophic methanogenic triculture. AB - Syntrophic degradation of normal- and branched-chain fatty acids with 4 to 9 carbons was investigated with a mesophilic syntrophic isobutyrate-butyrate degrading triculture consisting of the non-spore-forming, syntrophic, fatty acid degrading, gram-positive rod-shaped strain IB, Methanobacterium formicicum T1N, and Methanosarcina mazei T18. This triculture converted butyrate and isobutyrate to methane and converted valerate and 2-methylbutyrate to propionate and methane. This triculture also degraded caproate, 4-methylvalerate, heptanoate, 2 methylhexanoate, caprylate, and pelargoate. During the syntrophic conversion of isobutyrate and butyrate, a reversible isomerization between butyrate and isobutyrate occurred; isobutyrate and butyrate were isomerized to the other isomeric form to reach nearly equal concentrations and then their concentrations decreased at the same rates. Butyrate was an intermediate of syntrophic isobutyrate degradation. When butyrate was degraded in the presence of propionate, 2-methylbutyrate was synthesized from propionate and isobutyrate formed from butyrate. During the syntrophic degradation of valerate, isobutyrate, butyrate, and 2-methylbutyrate were formed and then degraded. During syntrophic degradation of 2-methylbutyrate, isobutyrate and butyrate were formed and then degraded. PMID- 16349312 TI - Cesium Accumulation and Growth Characteristics of Rhodococcus erythropolis CS98 and Rhodococcus sp. Strain CS402. AB - Growth and cesium accumulation characteristics of two cesium-accumulating bacteria isolated from soils were investigated. Rhodococcus erythropolis CS98 and Rhodococcus sp. strain CS402 accumulated high levels of cesium (approximately 690 and 380 mumol/g [dry weight] of cells or 92 and 52 mg/g [dry weight] of cells, respectively) after 24 h of incubation in the presence of 0.5 mM cesium. The optimum pH for cesium uptake by both Rhodococcus strains was 8.5. Rubidium and cesium assumed part of the role of potassium in the growth of both Rhodococcus strains. Potassium and rubidium inhibited cesium accumulation by these Rhodococcus strains. It is likely that both Rhodococcus strains accumulated cesium through a potassium transport system. PMID- 16349313 TI - Inoculum Density-Dependent Mortality and Colonization of the Phyllosphere by Pseudomonas syringae. AB - Pseudomonas syringae inocula containing cell concentrations ranging from 10 to 10 cells per ml were applied to the primary leaves of bean plants. The plants were incubated under conditions of high temperature and illumination and low relative humidity. Bacterial mortality rates and the proportional population decline of the inoculum were lowest at the highest inoculum concentrations. Addition of a high concentration of heat-killed cells to the inoculum containing a low concentration of viable cells significantly reduced both the mortality rate and the proportional population decline of the viable cells. The mechanisms underlying this density-dependent mortality may include cooperative protective effects of extracellular factors, such as bacterial extracellular polysaccharides, and physical protection by neighboring cells. Although epiphytic populations derived from inoculum concentrations of 10 or 10 cells per ml tended toward 10 CFU/g, the presumed carrying capacity of the leaf, populations derived from lower inoculum concentrations never achieved this carrying capacity. Assuming that epiphytic populations of P. syringae reside in discrete protected sites, our results suggest that at low inoculum concentrations, following a period of environmental stress, the number of viable cells may have dropped to zero in some sites; hence, the carrying capacity of the leaf could not be achieved. PMID- 16349314 TI - Multiple heavy metal tolerance of soil bacterial communities and its measurement by a thymidine incorporation technique. AB - A thymidine incorporation technique was used to determine the tolerance of a soil bacterial community to Cu, Cd, Zn, Ni, and Pb. An agricultural soil was artificially contaminated in our laboratory with individual metals at three different concentrations, and the results were compared with the results obtained by using the plate count technique. Thymidine incorporation was found to be a simple and rapid method for measuring tolerance. Data obtained by this technique were very reproducible. A linear relationship was found between changes in community tolerance levels obtained by the thymidine incorporation and plate count techniques (r = 0.732, P < 0.001). An increase in tolerance to the metal added to soil was observed for the bacterial community obtained from each polluted soil compared with the community obtained from unpolluted soil. The only exception was when Pb was added; no indication of Pb tolerance was found. An increase in the tolerance to metals other than the metal originally added to soil was also observed, indicating that there was multiple heavy metal tolerance at the community level. Thus, Cu pollution, in addition to increasing tolerance to Cu, also induced tolerance to Zn, Cd, and Ni. Zn and Cd pollution increased community tolerance to all five metals. Ni amendment increased tolerance to Ni the most but also increased community tolerance to Zn and, to lesser degrees, increased community tolerance to Pb and Cd. In soils polluted with Pb increased tolerance to other metals was found in the following order: Ni > Cd > Zn > Cu. We found significant positive relationships between changes in Cd, Zn, and Pb tolerance and, to a lesser degree, between changes in Pb and Ni tolerance when all metals and amendment levels were compared. The magnitude of the increase in heavy metal tolerance was found to be linearly related to the logarithm of the metal concentration added to the soil. Threshold tolerance concentrations were estimated from these linear relationships, and changes in tolerance could be detected at levels of soil contamination similar to those reported previously to result in changes in the phospholipid fatty acid pattern (A. Frostegard, A. Tunlid, and E. Baath, Appl. Environ. Microbiol. 59: 3605-3617, 1993). PMID- 16349315 TI - Rates of benthic protozoan grazing on free and attached sediment bacteria measured with fluorescently stained sediment. AB - In order to determine the importance of benthic protozoa as consumers of bacteria, grazing rates have been measured by using monodispersed fluorescently labeled bacteria (FLB). However, high percentages of nongrazing benthic protists are reported in the literature. These are related to serious problems of the monodispersed FLB method. We describe a new method using 5-(4,6-dichlorotriazin-2 yl)-aminofluorescein (DTAF)-stained sediment to measure in situ bacterivory by benthic protists. This method is compared with the monodispersed FLB technique. Our estimates of benthic bacterivory range from 61 to 73 bacteria protist h and are about twofold higher than the results of the monodispersed FLB method. The number of nongrazing protists after incubation for 15 min with DTAF-stained sediment is in agreement with theoretical expectation. We also tested the relative affinity for FLB of protists and discuss the results with respect to a grazing model. PMID- 16349316 TI - Derivation of Mutants of Erwinia carotovora subsp. betavasculorum Deficient in Export of Pectolytic Enzymes with Potential for Biological Control of Potato Soft Rot. AB - Erwinia carotovora subsp. betavasculorum Ecb168 produces an antibiotic(s) that suppresses growth of the related bacterium Erwinia carotovora subsp. carotovora in culture and in wounds of potato tubers. Strain Ecb168 also produces and secretes pectolytic enzymes and causes a vascular necrosis and root rot of sugar beet. Genes (out) involved in secretion of pectolytic enzymes by Ecb168 were localized to two HindIII fragments (8.5 and 10.5 kb) of Ecb168 genomic DNA by hybridization to the cloned out region of E. carotovora subsp. carotovora and by complementation of Out mutants of E. carotovora subsp. carotovora. Out mutants of Ecb168, which did not secrete pectate lyase into the culture medium, were obtained when deletions internal to either HindIII fragment were introduced into the genome of Ecb168 through marker exchange mutagenesis. Out mutants of Ecb168 were complemented to the Out phenotype by introduction of the corresponding cloned HindIII fragment. Out mutants of Ecb168 were less virulent than the Out parental strain on potato tubers. Strain Ecb168 and Out derivatives inhibited the growth of E. carotovora subsp. carotovora in culture, indicating that the uncharacterized antibiotic(s) responsible for antagonism was exported through an out-independent mechanism. Strain Ecb168 and Out derivatives reduced the establishment of large populations of E. carotovora subsp. carotovora in wounds of potato tubers and suppressed tuber soft rot caused by E. carotovora subsp. carotovora. PMID- 16349317 TI - Desulfonation of linear alkylbenzenesulfonate surfactants and related compounds by bacteria. AB - Pseudomonas putida S-313 (= DSM 6884) grew in sulfate-free medium when the sole sulfur source supplied was one of several arylsulfonates involved in the synthesis, application, or biodegradation of linear alkyl-benzenesulfonate (LAS) surfactants. 2-(4-Sulfophenyl)butyric acid, 4-n-butyl-1-methyl-6-sulfotetralin, and 4-toluenesulfonic acid were each completely utilized during growth, as were the model LAS 1-(4-sulfophenyl) octane and the arylsulfonate dyestuff Orange II. The product in each case was the corresponding phenol, which was identified by gas chromatography-mass spectrometry or H nuclear magnetic resonance. Stoichiometric conversion of 4-toluenesulfonic acid to 4-cresol was observed. The molar growth yields observed were 2.4 to 2.8 kg of protein per mol of S, which were comparable to the yield for sulfate. Commercial LAS disappeared from growth medium inoculated with strain S-313, but negligible growth occurred; digestion of cells in alkali led to recovery of the LAS mixture, which seemingly sorbed to the cells. However, mixed culture L6 was readily obtained from batch enrichment cultures containing commercial LAS as a sole sulfur source and an inoculum from domestic sewage. Culture L6 desulfonated components of the LAS surfactant to the corresponding phenols, which were identified by gas chromatography-mass spectrometry. Compounds with shorter alkyl chains were desulfonated preferentially, as were the centrally substituted isomers. In the presence of 200 muM sulfate, culture L6 grew well and LAS disappeared, although this was due purely to sorption, as shown by digestion of the cells in alkali. Thus, under sulfate-limited conditions, LAS can be desulfonated directly. PMID- 16349318 TI - Biocidal efficacy of a flocculating emergency water purification tablet. AB - Chlor-Floc (CF) emergency water purification tablets were tested for bactericidal, virucidal, and cysticidal efficacy in water at temperatures ranging from 5 to 25 degrees C. The minimal required log reduction was achieved for bacteria, Giardia muris, and rotavirus, but CF did not achieve the required log reduction of poliovirus at any of the temperatures or times investigated. The biocidal properties of the CF tablet were equivalent to if not greater than those of the Globaline iodine tablet, and the CF tablet was a more rapid cysticide under several potential use conditions. Therefore, it is a suitable substitute for iodine tablets for emergency purification of drinking water. Clarification of turbid waters was effective, but filtration through a cloth is necessary to prevent flocculated sediment from entering the canteen. The CF tablets met military requirements for emergency water purification and are safe and acceptable for use by the military. PMID- 16349319 TI - Carbon Metabolism Enzymes of Rhizobium tropici Cultures and Bacteroids. AB - We determined the activities of selected enzymes involved in carbon metabolism in free-living cells of Rhizobium tropici CFN299 grown in minimal medium with different carbon sources and in bacteroids of the same strain. The set of enzymatic activities in sucrose-grown cells suggests that the pentose phosphate pathway, with the participation of the Entner-Doudoroff pathway, is probably the primary route for sugar catabolism. In glutamate- and malate-grown cells, high activities of the gluconeogenic enzymes (phosphoenolpyruvate carboxykinase, fructose-6-phosphate aldolase, and fructose bisphosphatase) were detected. In bacteroids, isolated in Percoll gradients, the levels of activity for many of the enzymes measured were similar to those of malate-grown cells, except that higher activities of glucokinase, glucose-6-phosphate dehydrogenase, and NAD-dependent phosphogluconate dehydrogenase were detected. Phosphoglucomutase and UDP glucose pyrophosphorylase showed high and constant levels under all growth conditions and in bacteroids. PMID- 16349320 TI - Inhibitory Effects of Turf Pesticides on Bacillus popilliae and the Prevalence of Milky Disease. AB - Fourteen pesticides (fungicides, herbicides, and insecticides) were tested to determine whether they had deleterious effects on the bioinsecticide Bacillus popilliae, the causal agent of milky disease. All of these pesticides reduced levels of spore viability, spore germination, and/or vegetative cell growth when they were tested over a range of concentrations from 0 to 1,000 ppm of active ingredient, and the fungicides had the greatest detrimental effects. As determined by tests in water, the level of spore viability was significantly reduced by chlorothalonil, iprodione, (2,4-dichlorophenoxy)acetic acid plus 2 (2,4-dichlorophenoxy)propionic acid, and 2-[(4-chloro-o-tolyl)oxy]propionic acid plus (2,4-dichlorophenoxy)acetic acid. In tests performed with iprodione, loss of viability was evident at concentrations less than the concentration calculated to result from recommended use. Tests performed in soil demonstrated that triadimefon, chlorothalonil, (2,4-dichlorophenoxy)acetic acid plus 2-(2,4 dichlorophenoxy)propionic acid, and pendimethalin at concentrations resulting from recommended rates of application reduced spore titers. Spore germination did not occur in the continued presence of 2-[(4-chloro-otolyl)oxy]propionic acid plus (2,4-dichlorophenoxy)acetic acid, isofenphos, and chlordane, whereas exposure of spores to triadimefon or pendimethalin for 2 days stimulated germination. The tests to determine effects on spore germination were inconclusive for all other pesticides. Triadimefon, chlorothalonil, iprodione, pendimethalin, and chlorpyrifos at concentrations less than the concentrations recommended for use inhibited vegetative cell growth of B. popilliae, and chlordane at a concentration that was twice the concentration expected to result from the recommended rate of application repressed cell growth. My data support the hypothesis that use of synthetic pesticides can contribute to a low incidence of milky disease in white grubs. PMID- 16349321 TI - Uptake of Zinc in Pseudomonas sp. Strain UDG26. AB - Zinc resistance in Pseudomonas sp. strain UDG26 was inducible. Induction led to enhanced uptake of the metal. A zinc-sensitive variant (UDG86) took up significantly less metal ion than the resistant one did. The affinity of uninduced and sensitive cells to zinc was less than that of resistant, induced cells. Metal accumulation by induced cells was not inhibited by azide, while 2,4 dinitrophenol and N-N' -dicyclohexylcarbodiimide enhanced zinc uptake because of inhibition of efflux. Transcription and translation inhibitors drastically reduced zinc accumulation, bringing it to the level found in the sensitive strain. These results suggest the involvement of protein(s) in zinc resistance. PMID- 16349322 TI - Inheritance of Strain Instability (Sectoring) in the Commercial Button Mushroom, Agaricus bisporus. AB - The button mushroom, Agaricus bisporus, is a commercially important cultivated filamentous fungus. During the last decade, the button mushroom industry has depended mainly on two strains (or derivatives of these two strains). Using one of these highly successful strains (strain U1) we examined the phenomenon of strain instability, specifically, the production of irreversible sectors. Three "stromatal" and three "fluffy" sectors were compared with a healthy type U1 strain and with a wild-collected isolate. Compost colonization and fruit body morphology were examined. The main objective of this study, however, was to examine the meiotic stability of the sectored phenotype. Single basidiospores were isolated and subjected to a grain bioassay in which the ability to produce sectors was measured. Our results were as follows: (i) basidiospore cultures obtained from a wild-collected isolate showed no tendency to produce sectors; (ii) approximately 5% of the basidiospore cultures obtained from healthy type U1 strains produced irreversible sectors in the grain bioassay; (iii) the five primary sectors examined produced basidiospore cultures, half of which produced normal-looking growth in the grain bioassay and half of which produced some degree of sectoring; and (iv) the one sectored isolate that represented the F2 generation gave ratios similar to the 1:1 ratio observed for the F1 cultures. PMID- 16349323 TI - Novel processes for anaerobic sulfate production from elemental sulfur by sulfate reducing bacteria. AB - Sulfate reducers and related organisms which had previously been found to reduce Fe(III) with H(2) or organic electron donors oxidized S to sulfate when Mn(IV) was provided as an electron acceptor. Organisms catalyzing this reaction in washed cell suspensions included Desulfovibrio desulfuricans, Desulfomicrobium baculatum, Desulfobacterium autotrophicum, Desulfuromonas acetoxidans, and Geobacter metallireducens. These organisms produced little or no sulfate from S with Fe(III) as a potential electron acceptor or in the absence of an electron acceptor. In detailed studies with Desulfovibrio desulfuricans, the stoichiometry of sulfate and Mn(II) production was consistent with the reaction S + 3 MnO(2) + 4H-->SO(4) + 3Mn(II) + 2H(2)O. None of the organisms evaluated could be grown with S as the sole electron donor and Mn(IV) as the electron acceptor. In contrast to the other sulfate reducers evaluated, Desulfobulbus propionicus produced sulfate from S in the absence of an electron acceptor and Fe(III) oxide stimulated sulfate production. Sulfide also accumulated in the absence of Mn(IV) or Fe(III). The stoichiometry of sulfate and sulfide production indicated that Desulfobulbus propionicus disproportionates S as follows: 4S + 4H(2)O-->SO(4) + 3HS + 5 H. Growth of Desulfobulbus propionicus with S as the electron donor and Fe(III) as a sulfide sink and/or electron acceptor was very slow. The S oxidation coupled to Mn(IV) reduction described here provides a potential explanation for the Mn(IV)-dependent sulfate production that previous studies have observed in anoxic marine sediments. Desulfobulbus propionicus is the first example of a pure culture known to disproportionate S. PMID- 16349324 TI - Determination and Distribution of cry-Type Genes of Bacillus thuringiensis Isolates from Taiwan. AB - Using PCR with a set of specific oligonucleotide primers to detect cryI-type genes, we were able to screen the cry-type genes of 225 Bacillus thuringiensis soil isolates from Taiwan without much cost in time or labor. Some combinations of cry genes (the cry-type profile) in a single isolate were unique. We identified five distinct profiles of crystal genes from the B. thuringiensis soil isolates from Taiwan. The cry genes included cryIA(a), cryIA(b), cryIA(c), cryIC, cryID, and cryIV. Interestingly, 501 B. thuringiensis isolates (93.5% of the total number that we identified) were isolated from areas at high altitudes. The profiles of cry-type genes were distinct in all isolation areas. The distribution of cry-type genes of our isolates therefore depended on geography. Using PCR footprinting to detect cryIC-type genes, we identified two distinct cryIC footprints from some of our isolates, indicating that these isolates may contain novel cryIC-type genes. B. thuringiensis isolates containing cryIA(a)-, cryIA(b) , and cryIA(c)-type genes exhibited much greater activity against Plutella xylostella than did other isolates, indicating that multiple cry-type genes may be used as markers for the prediction of insecticidal activities. PMID- 16349325 TI - Effects of sieving, storage, and incubation temperature on the phospholipid Fatty Acid profile of a soil microbial community. AB - Disturbances typically associated with the study of soil microbial communities, i.e., sieving, storage, and subsequent incubation at elevated temperatures, were investigated with phospholipid fatty acid (PLFA) analyses. Treatment effects were quantified by statistical analyses of the mole percentage distribution of the individual fatty acids. Changes in the concentrations of individual fatty acids over a 7-week storage period at 4.5 degrees C were generally not statistically significant. Sieving effects (mesh size, 4 or 2 mm) on CO(2) evolution and the PLFA profile were monitored over 3 weeks; the physical disturbance had only minor effects, although some damage to fungal hyphae by the first sieving (<4 mm) was suggested by a decrease in the signature fatty acid 18:2 omega6c. Temperature effects were investigated by incubating soil for up to 3 weeks at 4.5, 10, or 25 degrees C. Principal component analyses demonstrated a significant shift in the PLFA composition at 25 degrees C over the first 2 weeks, while changes at the other two temperatures were minor. Several of the changes observed at 25 degrees C could be explained with reference to mechanisms of temperature adaptation or as a response to conditions of stress, including a decrease in the degree of unsaturation, an increased production of cyclopropyl fatty acids, and increased ratios of the branched-chain fatty acids iso-15:0 and iso-17:0 over anteiso-15:0 and anteiso-17:0, respectively. A decrease in the total amount of PLFA was also indicated. PMID- 16349326 TI - Determination of N Abundance in Nanogram Pools of NO(3) and NO(2) by Denitrification Bioassay and Mass Spectrometry. AB - Suspensions of two strains of Pseudomonas aeruginosa (ON12 and ON12-1) were used to reduce NO(3) and NO(2), respectively, to N(2)O. The evolved N(2)O was quantified by gas chromatography with electron capture detection, and the N abundance was determined by mass spectrometry with a special inlet system and triple-collector detection. Sample gas containing unknown N(2)O pools as small as 0.5 ng of N was analyzed by use of a spike technique, in which a reference gas of N(2)O of natural N abundance was added to obtain enough total N for the mass spectrometer. In NO(3) or NO(2) pools, the N abundance could be determined in samples as small as approximately 3.5 ng of N. No cross-contamination took place between the NO(3) and NO(2) pools. The excellent separation of NO(3) and NO(2) pools, small sample size required, and low contamination risk during N(2)O analysis offer great advantages in isotope studies of inorganic N transformations by, e.g., nitrifying or denitrifying bacteria in the environment. PMID- 16349327 TI - Accuracy, reproducibility, and interpretation of Fatty Acid methyl ester profiles of model bacterial communities. AB - We determined the accuracy and reproducibility of whole-community fatty acid methyl ester (FAME) analysis with two model bacterial communities differing in composition by using the Microbial ID, Inc. (MIDI), system. The biomass, taxonomic structure, and expected MIDI-FAME profiles under a variety of environmental conditions were known for these model communities a priori. Not all members of each community could be detected in the composite profile because of lack of fatty acid "signatures" in some isolates or because of variations (approximately fivefold) in fatty acid yield across taxa. MIDI-FAME profiles of replicate subsamples of a given community were similar in terms of fatty acid yield per unit of community dry weight and relative proportions of specific fatty acids. Principal-components analysis (PCA) of MIDI-FAME profiles resulted in a clear separation of the two different communities and a clustering of replicates of each community from two separate experiments on the first PCA axis. The first PCA axis accounted for 57.1% of the variance in the data and was correlated with fatty acids that varied significantly between communities and reflected the underlying community taxonomic structure. On the basis of our data, community fatty acid profiles can be used to assess the relative similarities and differences of microbial communities that differ in taxonomic composition. However, detailed interpretation of community fatty acid profiles in terms of biomass or community taxonomic composition must be viewed with caution until our knowledge of the quantitative and qualitative distribution of fatty acids over a wide variety of taxa and the effects of growth conditions on fatty acid profiles is more extensive. PMID- 16349328 TI - Kinetics of Colonization of Adult Queensland Fruit Flies (Bactrocera tryoni) by Dinitrogen-Fixing Alimentary Tract Bacteria. AB - The average total population of bacteria remained constant in the alimentary tracts of adult laboratory-raised Queensland fruit flies (Bactrocera tryoni) although the insects had ingested large numbers of live bacteria as part of their diet. The mean number of bacteria (about 13 million) present in the gut of the insects from 12 to 55 days after emergence was not significantly modified when, at 5 days after emergence, the flies were fed antibiotic-resistant bacteria belonging to two species commonly isolated from the gut of field-collected B. tryoni. Flies were fed one marked dinitrogen-fixing strain each of either Klebsiella oxytoca or Enterobacter cloacae, and the gastrointestinal tracts of fed flies were shown to be colonized within 7 days by antibiotic-resistant isolates of K. oxytoca but not E. cloacae. The composition of the microbial population also appeared to be stable in that the distribution and frequency of bacterial taxa among individual flies exhibited similar patterns whether or not the flies had been bacteria fed. Isolates of either E. cloacae or K. oxytoca, constituting 70% of the total numbers, were usually dominant, with oxidative species including pseudomonads forming the balance of the population. Antibiotic resistant bacteria could be spread from one cage of flies to the adjacent surfaces of a second cage within a few days and had reached a control group several meters distant by 3 weeks. Restriction of marked bacteria to the population of one in five flies sampled from the control group over the next 30 days suggested that the bacterial population in the gut of the insect was susceptible to alteration in the first week after emergence but that thereafter it entered a steady state and was less likely to be perturbed by the introduction of newly encountered strains. All populations sampled, including controls, included at least one isolate of the dinitrogen-fixing family Enterobacteriaceae; many were distinct from the marked strains fed to the flies. Nitrogenase activity detected by the acetylene reduction assay was associated with flies fed dinitrogen-fixing bacteria as well as with control groups given either no supplement or free access to a yeast hydrolysate preparation. Nitrogen fixed from the atmosphere may supplement the nutrition of the alimentary tract microbial population of B. tryoni. Transmission electron microscopy showed that the principal site of bacterial colonization in the abdominal alimentary tract was the lumen of the midgut inside the peritrophic membrane. No intracellular symbionts were seen in the gut tissues nor were bacteria found attached to the cuticular folds of the hindgut. The ultrastructure of the gut resembled that of other fly genera except that the intercellular spaces between rectal epithelial cells were more extensive, suggesting a role for unspecialized epithelium in water and solute uptake in B. tryoni. PMID- 16349329 TI - Oxidative Bioconversion of Cholesterol by Pseudomonas sp. Strain ST-200 in a Water-Organic Solvent Two-Phase System. AB - Pseudomonas sp. strain ST-200, which is capable of conversion of cholesterol, was isolated from humus soil. This organism effectively modified cholesterol dissolved in an organic solvent by dehydrogenation and oxygenation. When the organism was grown in a medium overlaid with a 10% volume of a mixed organic solvent (p-xylene and diphenylmethane; 3:7, vol/vol) containing cholesterol (20 mg/ml), the cholesterol concentration in the organic solvent was reduced to only 0.4 mg/ml after 8 days. Although the organism did not assimilate cholesterol, 98% of the cholesterol initially present disappeared. The organic solvent layer contained two major and three minor compounds converted from cholesterol. The major compounds were 6beta-hydroxycholest-4-en-3-one (8.9 mg/ml) and cholest-4 ene-3,6-dione (7.6 mg/ml). The concentrations of these compounds were equivalent to 43 and 37% of the cholesterol initially present. This organism would provide an effective and convenient system to oxidize the C-3 and -6 positions of cholesterol by introduction of a hydroxyl or ketone group. PMID- 16349330 TI - Pyranose Oxidase, a Major Source of H(2)O(2) during Wood Degradation by Phanerochaete chrysosporium, Trametes versicolor, and Oudemansiella mucida. AB - The production of the H(2)O(2)-generating enzyme pyranose oxidase (POD) (EC 1.1.3.10) (synonym, glucose 2-oxidase), two ligninolytic peroxidases, and laccase in wood decayed by three white rot fungi was investigated by correlated biochemical, immunological, and transmission electron microscopic techniques. Enzyme activities were assayed in extracts from decayed birch wood blocks obtained by a novel extraction procedure. With the coupled peroxidase-chromogen (3-dimethylaminobenzoic acid plus 3-methyl-2-benzothiazolinone hydrazone hydrochloride) spectrophotometric assay, the highest POD activities were detected in wood blocks degraded for 4 months and were for Phanerochaete chrysosporium (149 mU g [dry weight] of decayed wood), Trametes versicolor (45 mU g), and Oudemansiella mucida (1.2 mU g), corresponding to wood dry weight losses of 74, 58, and 13%, respectively. Mn-dependent peroxidase activities in the same extracts were comparable to those of POD, while lignin peroxidase activity was below the detection limit for all fungi with the veratryl alcohol assay. Laccase activity was high with T. versicolor (422 mU g after 4 months), in trace levels with O. mucida, and undetectable in P. chrysosporium extracts. Evidence for C-2 specificity of POD was shown by thin-layer chromatography detection of 2-keto-d glucose as the reaction product. By transmission electron microscopy immunocytochemistry, POD was found to be preferentially localized in the hyphal periplasmic space of P. chrysosporium and O. mucida and associated with membranous materials in hyphae growing within the cell lumina or cell walls of partially and highly degraded birch fibers. An extracellular distribution of POD associated with slime coating wood cell walls was also noted. The periplasmic distribution in hyphae and extracellular location of POD are consistent with the reported ultrastructural distribution of H(2)O(2)-dependent Mn-dependent peroxidases. This fact and the dominant presence of POD and Mn-dependent peroxidase in extracts from degraded wood suggest a cooperative role of the two enzymes during white rot decay by the test fungi. PMID- 16349331 TI - Dual Mechanisms of Tricarboxylate Transport and Catabolism by Acidaminococcus fermentans. AB - Acidaminococcus fermentans utilized citrate or the citrate analog aconitate as an energy source for growth, and these tricarboxylates were used simultaneously. Citrate utilization and uptake showed biphasic kinetics. High-affinity citrate uptake had a K(t) of 40 muM, but the V(max) was only 25 nmol/mg of protein per min. Low-affinity citrate utilization had a 10-fold higher V(max), but the K(s) was greater than 1.0 mM. Aconitate was a competitive inhibitor (K(i) = 34muM) of high-affinity citrate uptake, but low-affinity aconitate utilization had a 10 fold-lower requirement for sodium than did low-affinity citrate utilization. On the basis of this large difference in sodium requirements, it appeared that A. fermentans probably has two systems of tricarboxylate uptake: (i) a citrate/aconitate carrier with a low affinity for sodium and (ii) an aconitate carrier with a high affinity for sodium. Citrate was catabolized by a pathway involving a biotin-requiring, avidin-sensitive, sodium-dependent, membrane-bound oxaloacetate decarboxylase. The cells also had aconitase, but this enzyme was unable to convert citrate to isocitrate. Since cell-free extracts converted either aconitate or glutamate to 2-oxoglutarate, it appeared that aconitate was being catabolized by the glutaconyl-CoA decarboxylase pathway. Exponentially growing cultures on citrate or citrate plus aconitate were inhibited by the sodium/proton antiporter, monensin. Because monensin had no effect on cultures growing with aconitate alone, it appeared that citrate metabolism was acting as an inducer of monensin sensitivity. A. fermentans cells always had a low proton motive force (<50 mV), and cells treated with the protonophore TCS (3,3',4',5 tetrachlorosalicylanide) grew even though the proton motive force was less than 20 mV. On the basis of these results, it appeared that A. fermentans was depending almost exclusively on a sodium motive force for its membrane energetics. PMID- 16349332 TI - Contribution of the Global Regulator Gene gacA to Persistence and Dissemination of Pseudomonas fluorescens Biocontrol Strain CHA0 Introduced into Soil Microcosms. AB - Structural and regulatory genes involved in the synthesis of antimicrobial metabolites are essential for the biocontrol activity of fluorescent pseudomonads and, in principle, amenable to genetic engineering for strain improvement. An eventual large-scale release of such bacteria raises the question of whether such genes also contribute to the persistence and dissemination of the bacteria in soil ecosystems. Pseudomonas fluorescens wild-type strain CHA0 protects plants against a variety of fungal diseases and produces several antimicrobial metabolites. The regulatory gene gacA globally controls antibiotic production and is crucial for disease suppression in CHA0. This gene also regulates the production of extracellular protease and phospholipase. The contribution of gacA to survival and vertical translocation of CHA0 in soil microcosms of increasing complexity was studied in coinoculation experiments with the wild type and a gacA mutant which lacks antibiotics and some exoenzymes. Both strains were marked with spontaneous resistance to rifampin. In a closed system with sterile soil, strain CHA0 and the gacA mutant multiplied for several weeks, whereas these strains declined exponentially in nonsterile soil of different Swiss origins. The gacA mutant was less persistent in nonrhizosphere raw soil than was the wild type, but no competitive disadvantage when colonizing the rhizosphere and roots of wheat was found in the particular soil type and during the period studied. Vertical translocation was assessed after strains had been applied to undisturbed, long (60-cm) or short (20-cm) soil columns, both planted with wheat. A smaller number of cells of the gacA mutant than of the wild type were detected in the percolated water and in different depths of the soil column. Single-strain inoculation gave similar results in all microcosms tested. We conclude that mutation in a single regulatory gene involved in antibiotic and exoenzyme synthesis can affect the survival of P. fluorescens more profoundly in unplanted soil than in the rhizosphere. PMID- 16349333 TI - Purification and characterization of two versiconal hemiacetal acetate reductases involved in aflatoxin biosynthesis. AB - Two versiconal hemiacetal acetate (VHA) reductase activities (designated I and II), which catalyzed the reaction from VHA to versiconol acetate (VOAc) during aflatoxin biosynthesis, were purified to apparent homogeneity from the cytosol fraction of the mycelia of Aspergillus parasiticus mutant NIAH-26 through the following chromatography steps: first, fractionation with ammonium sulfate and then fractionation in succession with phenyl-Sepharose, DEAE-Sepharose, Sephacryl S-300, hydroxylapatite, and Matrex gel Green A chromatography. VHA reductase I and VHA reductase II were completely separated at the end of the DEAE-Sepharose step. The apparent molecular masses of reductase I and reductase II were estimated (by gel filtration) to be approximately 390 kDa; their denaturing molecular masses were 39- and 40-kDa, respectively (by sodium dodecyl sulfate polyacrylamide gel electrophoresis). Their pI values were 6.6 and 6.0, respectively (as determined by isoelectric focusing), and the optimal pH values were 8.0 and 9.0, respectively, although both enzymes exhibited a broad optimal pH range of between 7.5 and 9.0. The K(m) values of reductase I and reductase II for VHA were 35.4 and 25.4 muM, respectively. On the other hand, in the cell-free experiments involving either VHA reductase fraction and high-performance liquid chromatography, both (2'S)- and (2'R)-VOAc enantiomers were formed from racemic VHA and more of the 2'R isomer than the 2'S isomer was produced, indicating that the VHA reductase fractions have very similar stereospecificities to the substrate. PMID- 16349334 TI - Utilization of light for nitrogen fixation by a new synechocystis strain is extended by its low photosynthetic efficiency. AB - Performance of photosynthesis and nitrogenase activity in a novel cyanobacterium, Synechocystis sp. strain BO 8402, isolated from Lake Constance, located at the northern fringe of the Alps in central Europe, and of a stable derivative, strain BO 9201, were examined. Strain BO 8402 is characterized by an extraordinarily high level of autofluorescence originating from paracrystalline phycobiliprotein linker complexes located in inclusion bodies (W. Reuter, M. Westermann, S. Brass, A. Ernst, P. Boger, and W. Wehrmeyer, J. Bacteriol. 176:896-904, 1994). Energy transfer between paracrystalline phycobiliproteins and the photosystems is inefficient, resulting in a high oxygen compensation point and a decreased growth rate. The derivative strain BO 9201 exhibits hemidiscoidal phycobilisomes that support a high growth rate, even under low light intensities. Because of the differences in photosynthetic performance, anaerobic light-stimulated nitrogenase activity is maintained at higher light intensity in the original strain BO 8402 than in the derivative strain BO 9201. The results indicate that the formation of paracrystalline phycobiliproteins in Synechocystis sp. strain BO 8402 represents a hitherto-unknown means for a unicellular cyanobacterium to extend its capacity to fix nitrogen in the light. PMID- 16349335 TI - Isolation of a Bacterial Culture That Degrades Methyl t-Butyl Ether. AB - We have isolated a mixed bacterial culture (BC-1) which is capable of degrading the gasoline oxygenate methyl t-butyl ether (MTBE). BC-1 was developed from seed microorganisms present in a chemical plant biotreater sludge. This enrichment culture has been maintained in continuous culture treating high concentrations of MTBE (120 to 200 mg/liter) as the sole carbon source in a simple feed containing NH(4), PO(4), Mg, and Ca nutrients. The unit had a stable MTBE removal rate when maintained with a long cell retention time (ca. 80 to 90 days); however, when operated at a 6)/alpha(1-->3) glucosidic bonds. Cultures were mutagenized with UV and ethyl methanesulfonate, and colony morphology mutants were selected on 10% sucrose plates. Colony morphology variants exhibited changes from parent cultures in the production of one or more glucosyltransferases (GTFs) and glucans. Mutants were characterized by measuring resistance of glucan products to dextranase digestion, by electrophoresis, and by high-pressure liquid chromatography of maltose acceptor products generated from sucrose-maltose mixtures. Some mutants produced almost pure fraction L dextran, and cultures exhibited a single principal GTF band on sodium dodecyl sulfate-acrylamide gels. Other mutants produced glucans enriched for alternan. Colony morphology characteristics (size, smoothness, and opacity) and liquid culture properties (clumpiness, color, and viscosity in 10% sucrose medium) were explained on the basis of GTF production. Three principal GTF bands were detected. PMID- 16349347 TI - Bacterial Standing Stock, Activity, and Carbon Production during Formation and Growth of Sea Ice in the Weddell Sea, Antarctica. AB - Bacterial response to formation and growth of sea ice was investigated during autumn in the northeastern Weddell Sea. Changes in standing stock, activity, and carbon production of bacteria were determined in successive stages of ice development. During initial ice formation, concentrations of bacterial cells, in the order of 1 x 10 to 3 x 10 liter, were not enhanced within the ice matrix. This suggests that physical enrichment of bacteria by ice crystals is not effective. Due to low concentrations of phytoplankton in the water column during freezing, incorporation of bacteria into newly formed ice via attachment to algal cells or aggregates was not recorded in this study. As soon as the ice had formed, the general metabolic activity of bacterial populations was strongly suppressed. Furthermore, the ratio of [H]leucine incorporation into proteins to [H]thymidine incorporation into DNA changed during ice growth. In thick pack ice, bacterial activity recovered and growth rates up to 0.6 day indicated actively dividing populations. However, biomass-specific utilization of organic compounds remained lower than in open water. Bacterial concentrations of up to 2.8 x 10 cells liter along with considerably enlarged cell volumes accumulated within thick pack ice, suggesting reduced mortality rates of bacteria within the small brine pores. In the course of ice development, bacterial carbon production increased from about 0.01 to 0.4 mug of C liter h. In thick ice, bacterial secondary production exceeded primary production of microalgae. PMID- 16349348 TI - Genetic Structure of Rhizobium leguminosarum biovar trifolii and viciae Populations Found in Two Oregon Soils under Different Plant Communities. AB - An investigation was carried out to determine the genetic structure in soil populations of Rhizobium leguminosarum bv. trifolii and viciae at each of two Oregon sites (A and C) that were 1 km apart. Although the soils were similar, the plant communities were quite different because grazing by domestic animals had been allowed (site A) or prevented (site C). Analysis of allelic variation at 13 enzyme-encoding loci by multilocus enzyme electrophoresis delineated 202 chromosomal types (ETs) among a total of 456 isolates representing two populations of R. leguminosarum bv. trifolii (AT and CT) and two populations of R. leguminosarum bv. viciae (AV and CV). Regardless of their site of origin or biovar affiliation, isolates of the same ET were confirmed to be more closely related to each other than to isolates of other ETs by repetitive extragenic palindromic and enterobacterial repetitive intergeneric consensus sequences and the PCR technique. Despite the wide range in densities of the Rhizobium populations (<10 to >10/g of soil), their overall genetic diversities were similar (mean genetic diversity, 0.45 to 0.51), indicating that low-density populations of soil-borne bacterial species are not necessarily of little genetic diversity. Linkage disequilibrium analysis revealed significant multilocus structure (nonrandom associations of alleles) within each of the four populations. From a combination of cluster and linkage disequilibrium analyses, a total of eight distinct groups of ETs were defined in the four populations. Two groups (I and III) contributed significant numbers of ETs and isolates to each population. The two populations of R. leguminosarum bv. viciae (AV and CV) exhibited similar genetic structures despite existing at different densities, in different plant communities, and in the presence (CV) or absence (AV) of their local Vicia hosts. In contrast, total linkage disequilibrium was partitioned differently in two biovar populations occupying the same soil (AV and AT), with disequilibrium in the latter being due entirely to the presence of group V. PMID- 16349349 TI - Anisaldehyde and Veratraldehyde Acting as Redox Cycling Agents for H(2)O(2) Production by Pleurotus eryngii. AB - The existence of a redox cycle leading to the production of hydrogen peroxide (H(2)O(2)) in the white rot fungus Pleurotus eryngii has been confirmed by incubations of 10-day-old mycelium with veratryl (3,4-dimethoxybenzyl) and anisyl (4-methoxybenzyl) compounds (alcohols, aldehydes, and acids). Veratraldehyde and anisaldehyde were reduced by aryl-alcohol dehydrogenase to their corresponding alcohols, which were oxidized by aryl-alcohol oxidase, producing H(2)O(2). Veratric and anisic acids were incorporated into the cycle after their reduction, which was catalyzed by aryl-aldehyde dehydrogenase. With the use of different initial concentrations of either veratryl alcohol, veratraldehyde, or veratric acid (0.5 to 4.0 mM), around 94% of veratraldehyde and 3% of veratryl alcohol (compared with initial concentrations) and trace amounts of veratric acid were found when equilibrium between reductive and oxidative activities had been reached, regardless of the initial compound used. At concentrations higher than 1 mM, veratric acid was not transformed, and at 1.0 mM, it produced a negative effect on the activities of aryl-alcohol oxidase and both dehydrogenases. H(2)O(2) levels were proportional to the initial concentrations of veratryl compounds (around 0.5%), and an equilibrium between aryl-alcohol oxidase and an unknown H(2)O(2)-reducing system kept these levels steady. On the other hand, the concomitant production of the three above-mentioned enzymes during the active growth phase of the fungus was demonstrated. Finally, the possibility that anisaldehyde is the metabolite produced by P. eryngii for the maintenance of this redox cycle is discussed. PMID- 16349350 TI - Anaerobic degradation of propionate by a mesophilic acetogenic bacterium in coculture and triculture with different methanogens. AB - A mesophilic acetogenic bacterium (MPOB) oxidized propionate to acetate and CO(2) in cocultures with the formate- and hydrogen-utilizing methanogens Methanospirillum hungatei and Methanobacterium formicicum. Propionate oxidation did not occur in cocultures with two Methanobrevibacter strains, which grew only with hydrogen. Tricultures consisting of MPOB, one of the Methanobrevibacter strains, and organisms which are able to convert formate into H(2) plus CO(2) (Desulfovibrio strain G11 or the homoacetogenic bacterium EE121) also degraded propionate. The MPOB, in the absence of methanogens, was able to couple propionate conversion to fumarate reduction. This propionate conversion was inhibited by hydrogen and by formate. Formate and hydrogen blocked the energetically unfavorable succinate oxidation to fumarate involved in propionate catabolism. Low formate and hydrogen concentrations are required for the syntrophic degradation of propionate by MPOB. In triculture with Methanospirillum hungatei and the aceticlastic Methanothrix soehngenii, propionate was degraded faster than in biculture with Methanospirillum hungatei, indicating that low acetate concentrations are favorable for propionate oxidation as well. PMID- 16349351 TI - Three Native Cellulose-Depolymerizing Endoglucanases from Solid-Substrate Cultures of the Brown Rot Fungus Meruliporia (Serpula) incrassata. AB - Three extracellular cellulose-depolymerizing enzymes from cotton undergoing decay by the brown rot fungus Meruliporia (Serpula) incrassata were isolated by anion exchange and hydrophobic interaction chromatographies. Depolymerization was detected by analyzing the changes in the molecular size distribution of cotton cellulose by high-performance size-exclusion chromatography. The average degree of polymerization (DP; number of glucosyl residues per cellulose chain) was calculated from the size-exclusion chromatography data. The very acidic purified endoglucanases, Cel 25, Cel 49, and Cel 57, were glycosylated and had molecular weights of 25,200, 48,500, and 57,100, respectively. Two, Cel 25 and Cel 49, depolymerized cotton cellulose and were also very active on carboxymethyl cellulose (CMC). Cel 57, by contrast, significantly depolymerized cotton cellulose but did not release reducing sugars from CMC and only very slightly reduced the viscosity of CMC solutions. Molecular size distributions of cotton cellulose attacked by the three endoglucanases revealed single major peaks that shifted to lower DP positions. A second smaller peak (DP, 10 to 20) was also observed in the size-exclusion chromatograms of cotton attacked by Cel 49 and Cel 57. Under the reaction conditions used, Cel 25, the most active of the cellulases, reduced the weight average DP from 3,438 to 315, solubilizing approximately 20% of the cellulose. The weight average DP values of cotton attacked under the same conditions by Cel 49 and Cel 57 were 814 and 534; weight losses were 9 and 11% respectively. PMID- 16349352 TI - Partial Removal of Lipopolysaccharide from Thiobacillus ferrooxidans Affects Its Adhesion to Solids. AB - Conditions for the partial removal of lipopolysaccharide (LPS) from Thiobacillus ferrooxidans are described. Raising the pH of the solution containing the cells from pH 1.5 to pH 6.8 to 8.0 releases about 50% of the LPS without cell lysis. The release of LPS begins at pH 3.5, and it was not affected by EDTA concentration. Partial removal of LPS exposed higher amounts of a 40-kDa outer membrane protein in the bacteria, as detected by a dot immunoassay employing an antiserum against the T. ferrooxidans surface protein. This higher protein exposure and the reduced LPS content increased the hydrophobicity of the cell surface, as determined by an increased adhesion (50%) to hydrophobic sulfur prills and C-dodecanoic acid binding (2.5-fold) compared with control cells. In addition, adhesion of radioactively labeled microorganisms to a sulfide mineral was inhibited (40%) in the presence of previously added LPS. Our results suggest that not only LPS but also surface proteins probably play important roles in T. ferrooxidans adhesion to solid surfaces. PMID- 16349353 TI - Effect of Population Dynamics of Pseudomonas cepacia and Paecilomyces lilacinus on Colonization of Polyfoam Rooting Cubes by Rhizoctonia solani. AB - Suspensions of Pseudomonas cepacia (strain 5.5B) and Paecilomyces lilacinus (isolate 6.2F) were applied to polyfoam rooting cubes for control of stem rot of poinsettia caused by Rhizoctonia solani. The populations of antagonists and colonization of rooting cubes by R. solani were monitored during a 3-week period. Colonization of cubes by R. solani was reduced in cubes treated with P. cepacia, but the population of P. cepacia decreased by as much as 97% during the test period. Increased colonization by R. solani was correlated with a decline in population of P. cepacia. P. lilacinus was more persistent than P. cepacia in cubes, with only a 21% reduction observed during the 3-week period. Colonization of the P. lilacinus-treated cubes by R. solani was significantly less than colonization of infested controls. No correlation existed between population of P. lilacinus and colonization of cubes by R. solani. PMID- 16349354 TI - Chlorine Dioxide for Reduction of Postharvest Pathogen Inoculum during Handling of Tree Fruits. AB - Alternatives to hypochlorous acid and fungicides are needed for treatment of fruit and fruit-handling facilities. Chlorine dioxide was evaluated and found effective against common postharvest decay fungi and against filamentous fungi occurring on fruit packinghouse surfaces. In vitro tests with conidial or sporangiospore suspensions of Botrytis cinerea, Penicillium expansum, Mucor piriformis, and Cryptosporiopsis perennans demonstrated >99% spore mortality within 1 min when the fungi were exposed to aqueous chlorine dioxide at 3 or 5 mug . ml. Longer exposure times were necessary to achieve similar spore mortalities with 1 mug . ml. Of the fungi tested, B. cinerea and P. expansum were the least sensitive to ClO(2). In comparison with the number recovered from untreated control areas, the number of filamentous fungi recovered was significantly lower in swipe tests from hard surfaces such as belts and pads in a commercial apple and pear packinghouse after treatment of surfaces with a 14.0- to 18.0-mug . ml ClO(2) foam formulation. Chlorine dioxide has desirable properties as a sanitizing agent for postharvest decay management when residues of postharvest fungicides are not desired or allowed. PMID- 16349355 TI - Viability of indigenous soil bacteria assayed by respiratory activity and growth. AB - The bacterial population in barley field soil was estimated by determining the numbers of (i) cells reducing the artificial electron acceptor 5-cyano-2,3 ditolyl tetrazolium chloride (CTC) to CTC-formazan (respiratory activity), (ii) cells dividing a limited number of times (microcolony formation) on nutrient-poor media, (iii) cells dividing many times (colony formation) on nutrient-poor agar media, and (iv) cells stained with acridine orange (total counts). The CTC reduction assay was used for the first time for populations of indigenous soil bacteria and was further developed for use in this environment. The number of viable cells was highest when estimated by the number of microcolonies developing during 2 months of incubation on filters placed on the surface of nutrient-poor media. The number of bacteria reducing CTC to formazan was slightly lower than the number of bacteria forming microcolonies. Traditional plate counts of CFU (culturable cells) yielded the lowest estimate of viable cell numbers. The microcolony assay gave an estimate of both (i) cells forming true microcolonies (in which growth ceases after a few cell divisions) representing viable but nonculturable cells and (ii) cells forming larger microcolonies (in which growth continues) representing viable, culturable cells. The microcolony assay, allowing single-cell observations, thus seemed to be best suited for estimation of viable cell numbers in soil. The effect on viable and culturable cell numbers of a temperature increase from 4 to 17 degrees C for 5 days was investigated in combination with drying or wetting of the soil. Drying or wetting prior to the temperature increase, rather than the temperature increase per se, affected both the viable and culturable numbers of bacteria; both numbers were reduced in predried soil, while they increased slightly in the prewetted soil. PMID- 16349356 TI - Cloning and expression of genes required for coronamic Acid (2-ethyl-1 aminocyclopropane 1-carboxylic Acid), an intermediate in the biosynthesis of the phytotoxin coronatine. AB - Coronamic acid (CMA; 2-ethyl-1-aminocyclopropane 1-carboxylic acid) is an intermediate in the biosynthesis of coronatine (COR), a chlorosis-inducing phytotoxin produced by Pseudomonas syringae pv. glycinea PG4180. Tn5 mutagenesis and substrate feeding studies were previously used to characterize regions of the COR biosynthetic gene cluster required for synthesis of coronafacic acid and CMA, which are the only two characterized intermediates in the COR biosynthetic pathway. In the present study, additional Tn5 insertions were generated to more precisely define the region required for CMA biosynthesis. A new analytical method for CMA detection which involves derivatization with phenylisothiocyanate and detection by high-performance liquid chromatography (HPLC) was developed. This method was used to analyze and quantify the production of CMA by selected derivatives of P. syringae pv. glycinea which contained mutagenized or cloned regions from the CMA biosynthetic region. pMU2, a clone containing a 6.45-kb insert from the CMA region, genetically complemented mutants which required CMA for COR production. When pMU2 was introduced into P. syringae pv. glycinea 18a/90 (a strain which does not synthesize COR or its intermediates), CMA was not produced, indicating that pMU2 does not contain the complete CMA biosynthetic gene cluster. However, when two plasmid constructs designated pMU234 (12.5 kb) and pKTX30 (3.0 kb) were cointroduced into 18a/90, CMA was detected in culture supernatants by thin-layer chromatography and HPLC. The biological activity of the CMA produced by P. syringae pv. glycinea 18a/90 derivatives was demonstrated by the production of COR in cosynthesis experiments in which 18a/90 transconjugants were cocultivated with CMA-requiring mutants of P. syringae pv. glycinea PG4180. CMA production was also obtained when pMU234 and pKTX30 were cointroduced into P. syringae pv. syringae B1; however, these two constructs did not enable Escherichia coli K-12 to synthesize CMA. The production of CMA in P. syringae strains which lack the COR biosynthetic gene cluster indicates that CMA production can occur independently of coronafacic acid biosynthesis and raises interesting questions regarding the evolutionary origin of the COR biosynthetic pathway. PMID- 16349357 TI - Removal of Adsorbed Toxin Fragments That Modify Bacillus thuringiensis CryIC delta-Endotoxin Iodination and Binding by Sodium Dodecyl Sulfate Treatment and Renaturation. AB - We report that 10- and 25-kDa toxin fragments adhere to CryIC prepared from Bacillus thuringiensis insecticidal crystals, block iodination, and alter membrane binding. There is no apparent affect on CryIC toxicity against Spodoptera exigua. Associated peptides remained bound to CryIC in the presence of 50 mM dithiothreitol or 6 M urea. A novel detergent-renaturation procedure was developed for the purification of B. thuringiensis CryIC toxin. Sodium dodecyl sulfate (SDS) treatment followed by gel filtration chromatography yielded a homogeneous 62-kDa CryIC toxin. After removal of SDS and renaturation, the purified CryIC toxin was fully insecticidal to S. exigua larvae. I-labeled CryIC bound with high affinity to brush border membrane vesicles from S. exigua larvae. PMID- 16349358 TI - Factors Influencing the Synthesis of Polysaccharide by Bradyrhizobium japonicum Bacteroids in Field-Grown Soybean Nodules. AB - Certain strains of Bradyrhizobium japonicum produce large quantities of polysaccharide in soybean (Glycine max (L.) Merr.) nodules, and nodule polysaccharide (NPS) is different from that produced in culture. A previous survey of field-grown plants showed highly variable levels of NPS among field sites. To obtain clues about the possible function of NPS, we conducted two additional surveys of field-grown plants. The amount of polysaccharide in bulk samples of nodules was not associated with soil type, texture, slope, drainage, or any of the measured soil chemical properties except pH and [Ca]. Correlations with pH and [Ca] were positive and highly significant for two independent surveys involving a total of 77 sites in two years. In a preliminary comparison of high and low levels of Ca supplied to soybean plants grown in silica sand in a greenhouse, a high level of Ca (200 mg of Ca liter) increased the NPS level and increased the Ca content of the polysaccharide fraction. B. japonicum isolates from 450 nodules collected at 10 field sites in 1993 were used to form nodules on soybean plants grown in sand culture in a greenhouse in order to examine bacterial phenotype under controlled conditions. Results showed that the NPS level in the bulk nodule sample from any given site was a function of the proportion of nodule occupants that were capable of NPS synthesis. Thus, a higher soil pH and/or [Ca] may positively influence the survival of B. japonicum capable of synthesis of the nodule-specific polysaccharide. PMID- 16349359 TI - Culturability and Expression of Outer Membrane Proteins during Carbon, Nitrogen, or Phosphorus Starvation of Pseudomonas fluorescens DF57 and Pseudomonas putida DF14. AB - Changes in culturability and outer membrane protein profiles were investigated in Pseudomonas fluorescens DF57 and Pseudomonas putida DF14 during starvation for carbon, nitrogen, and phosphorus. P. fluorescens DF57 remained fully culturable for 4 days in all starvation regimes. The cell mass increased during starvation for nitrogen and phosphorus, indicating the accumulation of storage compounds, whereas it decreased slightly in carbon-starved cells. P. putida DF14 lost culturability during phosphorus starvation, and the mass of phosphate-starved cells did not increase. Analysis of additional P. fluorescens and P. putida strains, however, showed that the ability to preserve culturability during phosphorus starvation was not species but strain dependent. In DF57, an outer membrane protein of 55 kDa appeared during starvation for phosphorus, while another protein of 63 kDa was seen during all starvation conditions. DF14 induced two outer membrane proteins of 28 and 29 kDa during starvation for carbon and nitrogen, but no phosphorus-specific starvation protein could be detected. Therefore, starvation-induced outer membrane proteins do not seem to be conserved among the fluorescent pseudomonads and a unique starvation response might be found in individual strains. PMID- 16349360 TI - Cobalt(II) Oxidation by the Marine Manganese(II)-Oxidizing Bacillus sp. Strain SG 1. AB - The geochemical cycling of cobalt (Co) has often been considered to be controlled by the scavenging and oxidation of Co(II) on the surface of manganese [Mn(III,IV)] oxides or manganates. Because Mn(II) oxidation in the environment is often catalyzed by bacteria, we have investigated the ability of Mn(II)-oxidizing bacteria to bind and oxidize Co(II) in the absence of Mn(II) to determine whether some Mn(II)-oxidizing bacteria also oxidize Co(II) independently of Mn oxidation. We used the marine Bacillus sp. strain SG-1, which produces mature spores that oxidize Mn(II), apparently due to a protein in their spore coats (R.A. Rosson and K. H. Nealson, J. Bacteriol. 151:1027-1034, 1982; J. P. M. de Vrind et al., Appl. Environ. Microbiol. 52:1096-1100, 1986). A method to measure Co(II) oxidation using radioactive Co as a tracer and treatments with nonradioactive (cold) Co(II) and ascorbate to discriminate bound Co from oxidized Co was developed. SG-1 spores were found to oxidize Co(II) over a wide range of pH, temperature, and Co(II) concentration. Leucoberbelin blue, a reagent that reacts with Mn(III,IV) oxides forming a blue color, was found to also react with Co(III) oxides and was used to verify the presence of oxidized Co in the absence of added Mn(II). Co(II) oxidation occurred optimally around pH 8 and between 55 and 65 degrees C. SG-1 spores oxidized Co(II) at all Co(II) concentrations tested from the trace levels found in seawater to 100 mM. Co(II) oxidation was found to follow Michaelis Menten kinetics. An Eadie-Hofstee plot of the data suggests that SG-1 spores have two oxidation systems, a high-affinity-low-rate system (K(m), 3.3 x 10 M; V(max), 1.7 x 10 M . spore . h) and a low-affinity-high-rate system (K(m), 5.2 x 10 M; V(max), 8.9 x 10 M . spore . h). SG-1 spores did not oxidize Co(II) in the absence of oxygen, also indicating that oxidation was not due to abiological Co(II) oxidation on the surface of preformed Mn(III,IV) oxides. These results suggest that some microorganisms may directly oxidize Co(II) and such biological activities may exert some control on the behavior of Co in nature. SG-1 spores may also have useful applications in metal removal, recovery, and immobilization processes. PMID- 16349361 TI - Effect of Penicillium chrysogenum on Lignin Transformation. AB - A strain of Penicillium chrysogenum has been isolated from pine forest soils in Tenerife (Canary Islands). This strain was capable of utilizing hydroxylated and nonhydroxylated aromatic compounds, in particular cinnamic acid, as its sole carbon source. In an optimum medium with high levels of nitrogen (25.6 mM) and low levels of glucose (5.5 mM), it was able to decolorize Poly B-411 and to transform kraft, organosolv, and synthetic dehydrogenative polymerisate lignins. After 30 days of incubation, the amount of recovered kraft lignin was reduced to 83.5 and 91.3% of that estimated for uninoculated controls by spectrophotometry and klason lignin, respectively. At the same time, the pattern of molecular mass distribution of the lignin remaining in cultures was changed. The amount of organosolv lignin recovered from cultures was reduced to 90.1 and 94.6% of the initial amount as evaluated by spectrophotometry and klason lignin, respectively. About 6% of total applied radioactivity of OCH(3)-organosolv lignin was recovered as CO(2) after 30 days of incubation, and 18.5% of radioactivity from insoluble OCH(3)-organosolv lignin was solubilized. After 26 days of incubation, 2.9% of C beta-dehydrogenative polymerisate and 4.1% of C-ring-dehydrogenative polymerisate evolved as CO(2). PMID- 16349362 TI - Isolation, Growth, and Metabolism of an Obligately Anaerobic, Selenate-Respiring Bacterium, Strain SES-3. AB - A gram-negative, strictly anaerobic, motile vibrio was isolated from a selenate respiring enrichment culture. The isolate, designated strain SES-3, grew by coupling the oxidation of lactate to acetate plus CO(2) with the concomitant reduction of selenate to selenite or of nitrate to ammonium. No growth was observed on sulfate or selenite, but cell suspensions readily reduced selenite to elemental selenium (Se). Hence, SES-3 can carry out a complete reduction of selenate to Se. Washed cell suspensions of selenate-grown cells did not reduce nitrate, and nitrate-grown cells did not reduce selenate, indicating that these reductions are achieved by separate inducible enzyme systems. However, both nitrate-grown and selenate-grown cells have a constitutive ability to reduce selenite or nitrite. The oxidation of [C]lactate to CO(2) coupled to the reduction of selenate or nitrate by cell suspensions was inhibited by CCCP (carbonyl cyanide m-chlorophenylhydrazone), cyanide, and azide. High concentrations of selenite (5 mM) were readily reduced to Se by selenate-grown cells, but selenite appeared to block the synthesis of pyruvate dehydrogenase. Tracer experiments with [Se]selenite indicated that cell suspensions could achieve a rapid and quantitative reduction of selenite to Se. This reduction was totally inhibited by sulfite, partially inhibited by selenate or nitrite, but unaffected by sulfate or nitrate. Cell suspensions could reduce thiosulfate, but not sulfite, to sulfide. These results suggest that reduction of selenite to Se may proceed, in part, by some of the components of a dissimilatory system for sulfur oxyanions. PMID- 16349363 TI - Identification of Aeromonas species isolated from freshwater fish with the microplate hybridization method. AB - Aeromonas isolates were obtained from the intestinal tracts of six species of cultured freshwater fish and identified on the basis of their genotypic and phenotypic characters. The microplate hybridization method could differentiate type strains of Aeromonas species and related bacteria. DNA-DNA hybridization analysis showed that 65 aeromonad isolates were 72 to 100% related with either Aeromonas caviae, Aeromonas hydrophila, Aeromonas jandaei, Aeromonas sobria, or Aeromonas veronii. As many as 48% of the genotypically identified A. caviae, A. hydrophila, and A. sobria isolates differed from the type strains of corresponding species in one to three phenotypic characters. These results strongly suggest that not all aeromonad isolates from freshwater fish could be identified correctly on the basis of only the phenotypic characters, indicating the usefulness of the microplate hybridization method for the identification of aeromonads. PMID- 16349365 TI - Effects of ionic strength on bacterial adhesion and stability of flocs in a wastewater activated sludge system. AB - The success of biological wastewater treatment is to a large extent governed by the ability of bacteria to induce floc formation, thereby facilitating the separation of particles from the treated water. We performed studies on the dynamics of floc stability, the desorption of cells from the flocs, and the reflocculation of detached material. The floc stability was affected by the ionic strength of the medium in a way that strongly suggests that the interactions between the floc components can be explained by the theory of Derjaugin, Landau, Verwey, and Overbeek (DLVO theory). At increasing concentrations of electrolytes, the stability of the flocs increased. However, above an ionic strength of about 0.1 the floc stability decreased, and it seems that at this high electrolyte concentration the DLVO theory cannot be applied. The reversibility of the electrostatic double-layer effects was experimentally shown by treating the sludge repeatedly with a low-ionic-strength solution until parts of the flocs detached. When salt was added at this point, flocs re-form, resulting in a dramatic decrease in the turbidity of the supernatant liquid. Both reflocculation and detachment of floc material were seen with calcium as well as with potassium. This finding clearly indicates that the reflocculation and destabilization of flocs were due to changes in double-layer thickness rather than bridging effects of multivalent ions such as calcium. The results indicate that the ionic strength may well be an important factor for the floc stability in wastewater in situ. PMID- 16349366 TI - Inoculation of Scytalidium thermophilum in Button Mushroom Compost and Its Effect on Yield. AB - Scytalidium thermophilum isolates in culture, as well as the endogenous strain(s) in mushroom compost, were inactivated at 70 degrees C. This temperature was used to pasteurize composts for experiments. Of nine thermophilic fungal species, only S. thermophilum and Myriococcum thermophilum grew well on pasteurized compost in test tubes. The effect of both species on the crop yield of Agaricus bisporus mushrooms was studied. In solid-state fermentation rooms called tunnels, compost was pasteurized and inoculated. After incubation, the inoculated organisms were reisolated and counted, showing their successful colonization. The yield of mushrooms on inoculated composts was almost twice that on the pasteurized control. This result demonstrates the effectiveness of S. thermophilum in compost preparation. Inoculation is not necessary for traditional compost preparation. Naturally occurring strains of S. thermophilum, present in ingredients, readily colonize compost during preparation. Inoculation may be vital if compost is pretreated at a high temperature in tunnels. This finding is of relevance for the environmentally controlled production of high-yielding compost. PMID- 16349367 TI - Nuclear ribosomal DNA variation and pathogenic specialization in alternaria fungi known to produce host-specific toxins. AB - A total of 99 strains of 11 Alternaria species, including 68 strains of seven fungi known to produce host-specific toxins, were subjected to analysis of restriction fragment length polymorphism (RFLP) in nuclear ribosomal DNA (rDNA). Total DNA was digested with XbaI, and the Southern blots were probed with a nuclear rDNA clone of Alternaria kikuchiana. The hybridization gave 17 different RFLPs from the 99 strains. On the basis of these RFLPs, populations of host specific toxin-producing fungi could not be differentiated from one another nor from nonpathogenic A. alternata. Each population of the toxin-producing fungi carried rDNA variants. Nine different types, named A1 to A6 and B1 to B3, were detected among the toxin-producing fungi and nonpathogenic A. alternata. All of the populations contained the type A4 variant, and the other rDNA types were also shared by different toxin-producing fungi and A. alternata. In contrast, Alternaria species that are morphologically distinguishable from A. alternata could be differentiated from A. alternata on the basis of the rDNA RFLPs. Polymorphisms in rDNA digested with HaeIII and MspI were also evaluated in 61 Alternaria strains. These restriction enzymes produced 31 variations among all of the samples. The seven toxin-producing fungi and nonpathogenic A. alternata could not be resolved by phylogenetic analysis based on the RFLPs, although they could be differentiated from the other Alternaria species studied. These results provide support for the hypothesis that Alternaria fungi known to produce host specific toxins are intraspecific variants of A. alternata specialized in pathogenicity. PMID- 16349368 TI - Localization and Characterization of alpha-Glucosidase Activity in Lactobacillus brevis. AB - Lactobacillus brevis is found together with the yeast Brettanomyces lambicus during the overattenuation process in spontaneously fermented lambic beer. An isolated L. brevis strain has been shown to produce an alpha-glucosidase with many similarities to the glucosidase earlier found in B. lambicus. The enzyme was purified by ammonium sulfate precipitation, gel (Sephadex G-150 and Ultrogel AcA 44) filtration, and ion-exchange chromatography (DEAE-Sephadex A-50). The molecular weights of the enzyme, as determined by gel chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were about 50,000 and 60,000, respectively. Optimum catalytic activity was obtained at 40 degrees C and pH 6.0. The enzyme showed a decrease of hydrolysis with an increase in the degree of polymerization of the substrate. The K(m) values for p-nitrophenyl-alpha-d glucopyranoside, maltose, and maltotriose were 0.51, 3.0, and 5.2 mM, respectively. There was lack of inhibition by 0.15 mM acarbose and 0.5 M turanose, but the enzyme was inhibited by Tris (K(i) value of 25 mM). The alpha glucosidase of L. brevis together with the enzyme of B. lambicus seems to be a key factor in the overattenuation of lambic beer, although the involvement of other lactic acid bacteria (pediococci) cannot be excluded. PMID- 16349369 TI - Adsorption of Rhodococcus Strain GIN-1 (NCIMB 40340) on Titanium Dioxide and Coal Fly Ash Particles. AB - Rhodococcus strain GIN-1 (NCIMB 40340) can be used to enrich and isolate a titanium-rich fraction from coal fly ash. The gram-positive bacterium was isolated by its ability to adhere strongly and rapidly to suspended particles of pure titanium dioxide or coal fly ash. Adsorption depends on the salt concentration and occurs in seawater. Lowering of the salt concentration or washing of particles with pure water did not, however, cause desorption of the bacteria from TiO(2) particles; this was achieved by strong alkaline treatment or combined treatment with sodium dodecyl sulfate and urea but not with dilute acids, alcohols, or cationic or nonionic detergents. The bacterium exhibits higher affinity towards oxides of Ti and Zn than to other oxides with similar distribution of particle size. Moreover, it adheres much faster to TiO(2) than to magnetite (Fe(3)O(4)) or Al(2)O(3). After about 1 min, more than 85% of the cells were adsorbed on TiO(2), compared with adsorption of only 10 and 8% to magnetite and Al(2)O(3), respectively. Adsorption of the bacteria on TiO(2) occurs over a pH range of 1.0 to 9.0 and at temperatures from 4 to over 80 degrees C. Scanning electron microscopy combined with X-ray analysis revealed preferential adherence of the bacterium to coal ash particles richer in Ti. Stronger adhesion to TiO(2) was also demonstrated in the translocation of bacteria, preadsorbed on magnetite, to TiO(2) particles. The temporary co-adhesion to magnetite and TiO(2) was exploited for the design of a prototype biomagnetic separation process in which bacterial cells serve as an adhesive mediator between magnetite and TiO(2) particles in a mixture of Al, Si, and Ti oxides that simulates their proportion in the ash. PMID- 16349370 TI - Simulation Model of the Coupling between Nitrification and Denitrification in a Freshwater Sediment. AB - A model was constructed to simulate the results of experiments which investigated nitrification and denitrification in the freshwater sediment of Lake Vilhelmsborg, Denmark (K. Jensen, N. P. Sloth, N. Risgaard-Petersen, S. Rysgaard, and N. P. Revsbech, Appl. Environ. Microbiol. 60:2094-2100, 1994). The model output faithfully represented the profiles of O(2) and NO(3) and rates of nitrification, denitrification, and O(2) consumption as the O(2) concentration in the overlying water was increased from 10 to 600 muM. The model also accurately predicted the response, to increasing O(2) concentrations, of the integrated (micromoles per square meter per hour) rates of nitrification and denitrification. The simulated rates of denitrification of NO(3) diffusing from the overlying water (D(w)) and of NO(3) generated by nitrification within the sediment (D(n)) corresponded to the experimental rates as the O(2) concentration in the overlying water was altered. The predicted D(w) and D(n) rates, as NO(3) concentration in the overlying water was changed, closely resembled those determined experimentally. The model was composed of 41 layers 0.1 mm thick, of which 3 represented the diffusive boundary layer in the water. Large first-order rate constants for nitrification and denitrification were required to completely oxidize all NH(4) diffusing from the lower sediment layers and to remove much of the NO(3) produced. In addition to the flux of NH(4) from below, the model required a flux of an electron donor, possibly methane. Close coupling between nitrification and denitrification, achieved by allowing denitrification to tolerate some O(2) ( approximately 10 muM), was necessary to reproduce the real data. Spatial separation of the two processes (no toleration by denitrification of O(2)) resulted in too high NO(3) concentrations and too low rates of denitrification. PMID- 16349371 TI - Ecological Similarity and Coexistence of Epiphytic Ice-Nucleating (Ice) Pseudomonas syringae Strains and a Non-Ice-Nucleating (Ice) Biological Control Agent. AB - De Wit replacement series were used to study competitive interactions between epiphytic IcePseudomonas syringae strains and the biological frost control agents IceP. syringae TLP2del1 and Pseudomonas fluorescens A506. Mixtures containing two strains in different proportions but at a constant total population size were inoculated onto potato leaves. The population sizes of each strain and the total population size were determined when the community had reached equilibrium. A near-isogenic P. syringae strain pair exhibited an interaction similar to that expected for strains competing equally for limiting environmental resources. Replacement series with nonisogenic Ice and IceP. syringae strain pairs suggested that these strains competed for limiting resources according to their relative competitive abilities. There was no evidence of any niche differentiation between the IceP. syringae strains and the IceP. syringae strain. The growth responses of epiphytes following addition of nutrients to the phyllosphere indicated that the epiphytic P. syringae populations were nutrient limited and that, under growth chamber conditions, the populations were more limited by the availability of carbon than by the availability of nitrogen. Determination of in vitro carbon source utilization profiles provided further evidence for the lack of niche differentiation between the Ice and the IceP. syringae strains. Niche overlap indices calculated for the IceP. syringae strains with respect to IceP. syringae TLP2del1 were uniformly high, indicating ecological similarity, and were consistent with the observed low level of coexistence. The biological frost control agent P. fluorescens A506 replaced P. syringae. This was correlated with a high degree of niche overlap between these species. PMID- 16349372 TI - Dynamics and Distribution of Cyanophages and Their Effect on Marine Synechococcus spp. AB - Cyanophages infecting marine Synechococcus cells were frequently very abundant and were found in every seawater sample along a transect in the western Gulf of Mexico and during a 28-month period in Aransas Pass, Tex. In Aransas Pass their abundance varied seasonally, with the lowest concentrations coincident with cooler water and lower salinity. Along the transect, viruses infecting Synechococcus strains DC2 and SYN48 ranged in concentration from a few hundred per milliliter at 97 m deep and 83 km offshore to ca. 4 x 10 ml near the surface at stations within 18 km of the coast. The highest concentrations occurred at the surface, where salinity decreased from ca. 35.5 to 34 ppt and Synechococcus concentrations were greatest. Viruses infecting strains SNC1, SNC2, and 838BG were distributed in a similar manner but were much less abundant (<10 to >5 x 10 ml). When Synechococcus concentrations exceeded ca. 10 ml, cyanophage concentrations increased markedly (ca. 10 to > 10 ml), suggesting that a minimum host density was required for efficient viral propagation. Data on the decay rate of viral infectivity d (per day), as a function of solar irradiance I (millimoles of quanta per square meter per second), were used to develop a relationship (d = 0.2610I - 0.00718; r = 0.69) for conservatively estimating the destruction of infectious viruses in the mixed layer of two offshore stations. Assuming that virus production balances losses and that the burst size is 250, ca. 5 to 7% of Synechococcus cells would be infected daily by viruses. Calculations based on contact rates between Synechococcus cells and infectious viruses produce similar results (5 to 14%). Moreover, balancing estimates of viral production with contact rates for the farthest offshore station required that most Synechococcus cells be susceptible to infection, that most contacts result in infection, and that the burst size be about 324 viruses per lytic event. In contrast, in nearshore waters, where ca. 80% of Synechococcus cells would be contacted daily by infectious cyanophages, only ca. 1% of the contacts would have to result in infection to balance the estimated virus removal rates. These results indicate that cyanophages are an abundant and dynamic component of marine planktonic communities and are probably responsible for lysing a small but significant portion of the Synechococcus population on a daily basis. PMID- 16349373 TI - Hrp Mutants of Pseudomonas solanacearum as Potential Biocontrol Agents of Tomato Bacterial Wilt. AB - There have been many attempts to control bacterial wilt with antagonistic bacteria or spontaneous nonpathogenic mutants of Pseudomonas solanacearum that lack the ability to colonize the host, but they have met with limited success. Since a large gene cluster (hrp) is involved in the pathogenicity of P. solanacearum, we developed a biological control strategy using genetically engineered Hrp mutants of P. solanacearum. Three pathogenic strains collected in Guadeloupe (French West Indies) were rendered nonpathogenic by insertion of an omega-Km interposon within the hrp gene cluster of each strain. The resulting Hrp mutants were tested for their ability to control bacterial wilt in challenge inoculation experiments conducted either under growth chamber conditions or under greenhouse conditions in Guadeloupe. Compared with the colonization by a pathogenic strain which spread throughout the tomato plant, colonization by the mutants was restricted to the roots and the lower part of the stems. The mutants did not reach the fruit. Moreover, the presence of the mutants did not affect fruit production. When the plants were challenge inoculated with a pathogenic strain, the presence of Hrp mutants within the plants was correlated with a reduction in disease severity, although pathogenic bacteria colonized the stem tissue at a higher density than the nonpathogenic bacteria. Challenge inoculation experiments conducted under growth chamber conditions led, in some cases, to exclusion of the pathogenic strain from the aerial part of the plant, resulting in high protection rates. Furthermore, there was evidence that one of the pathogenic strains used for the challenge inoculations produced a bacteriocin that inhibited the in vitro growth of the nonpathogenic mutants. PMID- 16349374 TI - Minerals associated with biofilms occurring on exposed rock in a granitic underground research laboratory. AB - The concept of disposal of nuclear fuel waste in crystalline rock requires the effects of microbial action to be investigated. The Underground Research Laboratory excavated in a pluton of the Canadian Shield provides a unique opportunity to study these effects. Three biofilms kept moist by seepage through fractures in granitic rock faces of the Underground Research Laboratory have been examined. The biofilms contained a variety of gram-negative and gram-positive morphotypes held together by an organic extracellular matrix. Nutrient levels in the groundwater were low, but energy-dispersive X-ray spectroscopy has shown biogeochemical immobilization of several elements in the biofilms; some of these elements were concentrated from extremely dilute environmental concentrations, and all elements were chemically complexed together to form amorphous or crystalline fine-grained minerals. These were seen by transmission electron microscopy to be both associated with the surfaces of the bacteria and scattered throughout the extracellular matrix, suggesting their de novo development through bacterial surface-mediated nucleation. The biofilm consortia are thought to concentrate elements both by passive sorption and by energy metabolism. By Mossbauer spectroscopy and X-ray diffraction, one of the biofilms showed that iron was both oxidized and precipitated as ferrihydrite or hematite aerobically and reduced and precipitated as siderite anaerobically. We believe that some Archean banded-iron formations could have been formed in a manner similar to this, as it would explain the deposition of hematite and siderite in close proximity. This biogeochemical development of minerals may also affect the transport of material in waste disposal sites. PMID- 16349375 TI - Microscopic and thermal characterization of hydrogen peroxide killing and lysis of spores and protection by transition metal ions, chelators, and antioxidants. AB - Killing of bacterial spores by H(2)O(2) at elevated but sublethal temperatures and neutral pH occurred without lysis. However, with prolonged exposure or higher concentrations of the agent, secondary lytic processes caused major damage successively to the coat, cortex, and protoplast, as evidenced by electron and phase contrast microscopy. These processes were also reflected in changes in differential scanning calorimetric profiles for H(2)O(2)-treated spores. Endothermic transitions in the profiles occurred at lower temperatures than usual as a result of H(2)O(2) damage. Thus, H(2)O(2) sensitized the cells to heat damage. Longer exposure to H(2)O(2) resulted in total disappearance of the transitions, indicative of major disruptions of cell structure. Spores but not vegetative cells were protected against the lethal action of H(2)O(2) by the transition metal cations Cu, Cu, Co, Co, Fe, Fe, Mn, Ti, and Ti. The metal chelator EDTA was also somewhat protective, while o-phenanthroline, citrate, deferoxamine, and ethanehydroxydiphosphonate were only marginally so. Superoxide dismutase and a variety of other free-radical scavengers were not protective. In contrast, reducing agents such as sulfhydryl compounds and ascorbate at concentrations of 20 to 50 mM were highly protective. Decoating or demineralization of the spores had only minor effects. The marked dependence of H(2)O(2) sporicidal activity on moderately elevated temperature and the known low reactivity of H(2)O(2) itself suggest that radicals are involved in its killing action. However, the protective effects of a variety of oxidized or reduced transition metal ions indicate that H(2)O(2) killing of spores is markedly different from that of vegetative cells. PMID- 16349376 TI - Lactococcal bacteriophages require a host cell wall carbohydrate and a plasma membrane protein for adsorption and ejection of DNA. AB - The mechanism of the initial steps of bacteriophage infection in Lactococcus lactis subsp. lactis C2 was investigated by using phages c2, ml3, kh, l, h, 5, and 13. All seven phages adsorbed to the same sites on the host cell wall that are composed, in part, of rhamnose. This was suggested by rhamnose inhibition of phage adsorption to cells, competition between phage c2 and the other phages for adsorption to cells, and rhamnose inhibition of lysis of phage-inoculated cultures. The adsorption to the cell wall was found to be reversible upon dilution of the cell wall-adsorbed phage. In a reaction step that apparently follows adsorption to the cell wall, all seven phages adsorbed to a host membrane protein named PIP. This was indicated by the inability of all seven phages to infect a strain selected for resistance to phage c2 and known to have a defective PIP protein. All seven phages were inactivated in vitro by membranes from wild type cells but not by membranes from the PIP-defective, phage c2-resistant strain. The mechanism of membrane inactivation was an irreversible adsorption of the phage to PIP, as indicated by adsorption of [S] methionine-labeled phage c2 to purified membranes from phage-sensitive cells but not to membranes from the resistant strain, elimination of adsorption by pretreatment of the membranes with proteinase K, and lack of dissociation of S from the membranes upon dilution. Following membrane adsorption, ejection of phage DNA occurred rapidly at 30 degrees C but not at 4 degrees C. These results suggest that many lactococcal phages adsorb initially to the cell wall and subsequently to host cell membrane protein PIP, which leads to ejection of the phage genome. PMID- 16349377 TI - Identification of Whole Fixed Bacterial Cells with Nonradioactive 23S rRNA Targeted Polynucleotide Probes. AB - Polyribonucleotide probes (ca. 200 to 300 nucleotides in length) carrying multiple reporter molecules were produced by in vitro transcription with labeled UTP derivatives (fluorescein-12-UTP, 7-amino-4-methyl-coumarin-3-acetyl-6-UTP, tetramethylrhodamine-6-UTP, or digoxigenin-11-UTP). Despite their length, these molecules penetrated into whole fixed gram-negative cells and hybridized specifically to their target sites on the 23S rRNA. Fluorescence intensities were quantified for target and nontarget cells by the combination of a charge-coupled device videocamera and an image-processing system. Polyribonucleotide probes confer up to 26 times more fluorescence to target cells than oligonucleotide probes do. Probe sensitivity and specificity were strongly influenced by the stringency of hybridization. The use of differently labeled probes allowed the simultaneous detection of three populations. Identification of introduced test organisms in activated-sludge samples proved the applicability of this method for the in situ identification of microorganisms in complex microbial communities. PMID- 16349378 TI - Polyester Biosynthesis Characteristics of Pseudomonas citronellolis Grown on Various Carbon Sources, Including 3-Methyl-Branched Substrates. AB - Forty-two different carbon sources were tested for the polyester synthesis of a citronellol-utilizing bacterium, Pseudomonas citronellolis (ATCC 13674). These included linear C(2) to C(10) monocarboxylic acids, C(3) to C(10) dicarboxylic acids, saccharides, alpha,omega-diols, hydrocarbons, and 3-methyl-branched substrates such as 3,7-dimethyl-6-octen-1-ol (citronellol), 3-methyl-n-valerate, 3-methyl-1-butanol, and 3-methyladipate. Isolated polymers were characterized by gas chromatography, infrared spectroscopy, H- or C-nuclear magnetic resonance spectroscopy, H-C heteronuclear correlation spectroscopy (H-C COSY), H-H homonuclear COSY, and differential scanning calorimetry. Polyesters from nine monocarboxylic acids and two related carbon sources could be metabolically divided into three groups. The first group of C(2) to C(4) carbon sources resulted in copolyesters composed of 61 to 70 mol% 3-hydroxydecanoate, 23 to 33 mol% 3-hydroxyoctanoate, 3.6 to 9.0 mol% 3-hydroxy-5-cis-dodecenoate, and 1.8 to 2.6 mol% 3-hydroxy-7-cis-tetradecenoate. Carbon sources in group II (C(7) to C(10)) produced copolyesters composed of 3-hydroxyacid monomer units with the same number of carbon atoms as the substrate (major constituent) and monomer units with either two less or two more carbons. Negligible amounts of 3-hydroxy-5 cis-dodecenoate and 3-hydroxy-7-cis-tetradecenoate were detected in copolyesters from this group. Copolyesters from group III (C(5) and C(6)) had a monomer unit distribution that could be said to be between those of groups I and II. In addition, a novel copolyester, poly(3-hydroxy-7-methyl-6-octenoate-co-3-hydroxy-5 methylhexanoate), was synthesized when grown on citronellol. The H-C heteronuclear COSY spectrum for monomer unit II revealed that both methylene and isopropyl groups, proximately connected in series to a single chiral center, had magnetically diastereotopic natures. PMID- 16349379 TI - Oxidative dissolution of arsenopyrite by mesophilic and moderately thermophilic acidophiles. AB - The purpose of this work was to determine solution- and solid-phase changes associated with the oxidative leaching of arsenopyrite (FeAsS) by Thiobacillus ferrooxidans and a moderately thermoacidophilic mixed culture. Jarosite [KFe(3)(SO(4))(2)(OH)(6)], elemental sulfur (S), and amorphous ferric arsenate were detected by X-ray diffraction as solid-phase products. The oxidation was not a strongly acid-producing reaction and was accompanied by a relatively low redox level. The X-ray diffraction lines of jarosite increased considerably when ferrous sulfate was used as an additional substrate for T. ferroxidans. A moderately thermoacidophilic mixed culture oxidized arsenopyrite faster at 45 degrees C than did T. ferroxidans at 22 degrees C, and the oxidation was accompanied by a nearly stoichiometric release of Fe and As. The redox potential was initially low but subsequently increased during arsenopyrite oxidation by the thermoacidophiles. Jarosite, S, and amorphous ferric arsenate were also formed under these conditions. PMID- 16349380 TI - Relationship between Phylogeny and Pathotype for the Bacterial Blight Pathogen of Rice. AB - Several transposable elements were isolated from the genome of Xanthomonas oryzae pv. oryzae. These elements and an avirulence gene isolated from X. oryzae pv. oryzae were used as hybridization probes for a collection of X. oryzae pv. oryzae strains from the Philippines. Each of the sequences was present in multiple copies in all strains examined and showed distinct patterns of hybridizing bands. Phenograms were derived from the restriction fragment length polymorphism data obtained for each of the individual probes and for pooled data from multiple probes. The phenograms derived from the different probes differed in topology and, on the basis of bootstrap analysis, were not equally robust. For all of the probes, including the avirulence gene, some groups (even some haplotypes) consisted of multiple races. The strains were grouped into four major clusters on the basis of the two probes giving the highest bootstrap values. These groups were inferred to represent phylogenetic lineages. Three of the six races of X. oryzae pv. oryzae appeared in more than one of the lineages, and another was present in two sublineages. For three of the races, strains representing different phenetic groups were inoculated on rice cultivars carrying 10 resistance genes. Two new races were differentiated, corresponding to pathogen lineages identified by DNA typing. On the basis of DNA and pathotypic analyses, together with information on the spatial and temporal distribution of the pathogen types from this and other studies, a general picture of X. oryzae pv. oryzae evolution in the Philippines is presented. PMID- 16349381 TI - Influence of Viable Cells on the Resuscitation of Dormant Cells in Micrococcus luteus Cultures Held in an Extended Stationary Phase: the Population Effect. AB - A high proportion of Micrococcus luteus cells in cultures which had been starved for 3 to 6 months lost the ability to grow and form colonies on agar plates but could be resuscitated from their dormancy by incubation in an appropriate liquid medium (A. S. Kaprelyants and D. B. Kell, Appl. Environ. Microbiol. 59:3187-3196, 1993). In the present work, such cultures were studied by both flow cytometry and conventional microbiological methods and were found to contain various numbers of viable cells. Pretreatment of such cultures with penicillin G, and subsequent dilution, was used to vary this number. When the initial number of colony-forming cells per 30-ml flask was approximately nine (+/-five) or more, resuscitation of 10 to 40% of the cells, and thus culture growth, was observed. The lag period before the appearance of a population of cells showing significant accumulation of the fluorescent dye rhodamine 123 (i.e., of cells with measurable membrane energization) decreased from 70 to 27 h when the number of viable cells was increased from 30 to 10 per flask, while the lag period before an observable increase in the number of colony-forming cells occurred was almost constant (at some 20 h). Provided there were more than nine (+/-five) initially viable cells per flask, the number of initially viable cells did not affect the final percentage of resuscitable cells in the culture. The lag period could be ascribed in part to the time taken to restore the membrane permeability barrier of starved cells during resuscitation, as revealed by flow cytometric assessment of the uptake of the normally membrane-impermeant fluorescent DNA stain PO-PRO-3 {4-[3 methyl-2, 3-dihydro-(benzo-1, 3-oxazole)-2-methylidene]-1-(3'-trimethylammonium propyl)-pyridinium diiodide}. Although cell populations which contained fewer than nine +/-five viable cells per flask failed to grow, 4 to 20% of the cells (of 1.2 X 10) were able to accumulate rhodamine 123 after 80 to 100 h of incubation, showing the ability of a significant number of the cells in the population at least to display "metabolic resuscitation." Resuscitation and cell growth under such conditions were favored by the use of a 1:1 mixture of fresh lactate medium and supernatant from late-logarithmic-phase M. luteus cultures as the resuscitation medium. We conclude that the presence of a small fraction of viable cells at the onset of resuscitation facilitates the recovery of the majority of the remaining (dormant) cells. The cell density dependence of the kinetics, or population effect, suggests that this recovery is due to the excretion of some factor(s) which promoted the transition of cells from a state in which they are incapable of growth and division to one in which they are capable of colony formation. PMID- 16349382 TI - Changes in Ester-Linked Phospholipid Fatty Acid Profiles of Subsurface Bacteria during Starvation and Desiccation in a Porous Medium. AB - Ester-linked phospholipid fatty acid (PLFA) profiles of a Pseudomonas aureofaciens strain and an Arthrobacter protophormiae strain, each isolated from a subsurface sediment, were quantified in a starvation experiment in a silica sand porous medium under moist and dry conditions. Washed cells were added to sand microcosms and maintained under saturated conditions or subjected to desiccation by slow drying over a period of 16 days to final water potentials of approximately - 7.5 MPa for the P. aureofaciens and - 15 MPa for the A. protophormiae. In a third treatment, cells were added to saturated microcosms along with organic nutrients and maintained under saturated conditions. The numbers of culturable cells of both bacterial strains declined to below detection level within 16 days in both the moist and dried nutrient-deprived conditions, while direct counts and total PLFAs remained relatively constant. Both strains of bacteria maintained culturability in the nutrient-amended microcosms. The dried P. aureofaciens cells showed changes in PLFA profiles that are typically associated with stressed gram-negative cells, i.e., increased ratios of saturated to unsaturated fatty acids, increased ratios of trans- to cis-monoenoic fatty acids, and increased ratios of cyclopropyl fatty acids to their monoenoic precursors. P. aureofaciens starved under moist conditions showed few changes in PLFA profiles during the 16-day incubation, whereas cells incubated in the presence of nutrients showed decreases in the ratios of both saturated fatty acids to unsaturated fatty acids and cyclopropyl fatty acids to their monoenoic precursors. The PLFA profiles of A. protophormiae changed very little in response to either nutrient deprivation or desiccation. Diglyceride fatty acids, which have been proposed to be indicators of dead or lysed cells, remained relatively constant throughout the experiment. Only the A. protophormiae desiccated for 16 days showed an increase in the ratio of diglyceride fatty acids to PLFAs. The results of this laboratory experiment can be useful for interpreting PLFA profiles of subsurface communities of microorganisms for the purpose of determining their physiological status. PMID- 16349383 TI - Effect of solution ionic strength and iron coatings on mineral grains on the sorption of bacterial cells to quartz sand. AB - Understanding the interaction between bacterial cells and solid surfaces is essential to our attempts to quantify and predict the transport of microbes in groundwater aquifers, whether from the point of view of contamination or from that of bioremediation. The sorption of bacterial cells suspended in groundwater to porous medium grains was examined in batch studies. Bacterial sorption to clean quartz sand yielded equilibrium, linear, adsorption isotherms that varied with the bacterial strain used and the ionic strength of the aqueous solution. Values of K(d) (the slope of the linear sorption isotherm) ranged from 0.55 to 6.11 ml g, with the greatest sorption observed for the highest groundwater ionic strength. These findings are consistent with the interpretation that an increasingly compressed electrical double layer results in stronger adsorption between the like-charged mineral surface and the bacterial cells. When iron oxyhydroxide-coated sand was used, however, all of the added bacteria were adsorbed up to a threshold of 6.93 x 10 cells g of coated sand, beyond which no further adsorption occurred. The irreversible, threshold adsorption is the result of a strong electrostatic attraction between the sesquioxide coating and the bacterial cells. Experimental results of adsorption in mixtures of quartz and Fe(III)-coated sand were successfully predicted by a simple additive model for sorption by the two substrate phases. Even small amounts of Fe(III)-coated sand in a mixture influenced the extent of adsorption of bacterial cells. A quantitative description of adsorption in the mixtures can be realized by using a linear isotherm for reversible adsorption to the quartz grains with a y intercept that represents the number of cells irreversibly adsorbed to the Fe(III)-coated sand. PMID- 16349384 TI - Effects of methane metabolism on nitrification and nitrous oxide production in polluted freshwater sediment. AB - We report the effect of CH(4) and of CH(4) oxidation on nitrification in freshwater sediment from Hamilton Harbour, Ontario, Canada, a highly polluted ecosystem. Aerobic slurry experiments showed a high potential for aerobic N(2)O production in some sites. It was suppressed by C(2)H(2), correlated to NO(3) production, and stimulated by NH(4) concentration, supporting the hypothesis of a nitrification-dependent source for this N(2)O production. Diluted sediment slurries supplemented with CH(4) (1 to 24 muM) showed earlier and enhanced nitrification and N(2)O production compared with unsupplemented slurries (500 ppm in the presence of 250 muM nitrite. Inhibition increased with increasing ammonium at methane concentrations of 100 ppm. In the presence of 500 muM ammonium, inhibition increased initially with increasing methane concentrations from 1.7 to 100 ppm; the extent of inhibition decreased with methane concentrations of > 100 ppm. The results of this study provide new insights that explain some of the previously observed interactions among ammonium, nitrite, methane, and methane oxidation in soils and aquatic systems. PMID- 16349403 TI - Mechanistic analysis of ammonium inhibition of atmospheric methane consumption in forest soils. AB - Methane consumption by forest soil was studied in situ and in vitro with respect to responses to nitrogen additions at atmospheric and elevated methane concentrations. Methane concentrations in intact soil decreased continuously from atmospheric levels at the surface to 0.5 ppm at a depth of 14 cm. The consumption rate of atmospheric methane in soils, however, was highest in the 4- to 8-cm depth interval (2.9 nmol per g of dry soil per day), with much lower activities below and above this zone. In contrast, extractable ammonium and nitrate concentrations were highest in the surface layer (0 to 2 cm; 22 and 1.6 mumol per g of dry soil, respectively), as was potential ammonium-oxidizing activity (19 nmol per g of dry soil per day). The difference in zonation between ammonium oxidation and methane consumption suggested that ammonia-oxidizing bacteria did not contribute significantly to atmospheric methane consumption. Exogenous ammonium inhibited methane consumption in situ and in vitro, but the pattern of inhibition did not conform to expectations based on simple competition between ammonia and methane for methane monooxygenase. The extent of ammonium inhibition increased with increasing methane concentration. Inhibition by a single ammonium addition remained constant over a period of 39 days. In addition, nitrite, the end product of methanotrophic ammonia oxidation, was a more effective inhibitor of methane consumption than ammonium. Factors that stimulated ammonium oxidation in soil, e.g., elevated methane concentrations and the availability of cosubstrates such as formate, methanol, or beta-hydroxybutyrate, enhanced ammonium inhibition of methane oxidation, probably as a result of enhanced nitrite production. PMID- 16349404 TI - Effects of Neem Leaf Volatiles on Submerged Cultures of Aflatoxigenic Aspergillus parasiticus. AB - Microbe-free compressed air was passed continuously for a 3-day test period through an enclosed system containing fresh neem leaves; the resultant emitted volatiles were passed over the surface of submerged liquid cultures of a wild type aflatoxigenic isolate of Aspergillus parasiticus. Aflatoxin determinations for the fungal culture that received neem-derived volatiles, after a 3-day incubation period, resulted in a 90% overall reduction in aflatoxin production and a 51% reduction in fungal biomass when compared with cultures that did not receive neem volatiles. In a separate experiment but in a similarly enclosed system, volatiles from fresh neem leaves were collected on a small Tenax column and were thermally desorbed and cryogenically focused on a capillary gas chromatography column. The neem volatiles were subsequently separated and identified by gas chromatography-mass spectrometry. Sixty-eight compounds were identified by comparison of retention times and mass spectra with either authentic compounds or spectra from a computer-assisted library database of mass spectra. It was found that 10% of the total headspace volatiles were composed of C(3) to C(9) alkenals, which are toxic to aflatoxigenic Aspergillus spp., which could explain the bioactivity that resulted in reduced biomass in the neem treated cultures. PMID- 16349405 TI - A Sensitive Method Using 4-Methylumbelliferyl-beta-Cellobiose as a Substrate To Measure (1,4)-beta-Glucanase Activity in Sediments. AB - A sensitive method to measure (1,4)-beta-glucanase activity in organic matter rich sediments, using 4-methyl-umbelliferyl-beta-cellobiose as a substrate, is described. beta-Glucosidases, which were also able to hydrolyze this substrate, were inhibited with d-glucono-delta-lactone. The produced 4-methylumbelliferone was recovered quantitatively out of the sediment by an extraction with 80% ethanol. An inhibition experiment with known substrates or inhibitors suggested that at least 59% of the measured activity could be explained by enzymes of the exo-(1,4)-beta-glucanase type and that the contribution of endo-(1,4)-beta glucanases was minor. Results of the inhibition experiment also suggested that the measured activity was of bacterial origin in the sediment used. First results of field measurements are given for the sediment from the reed bed of Lake Gooimeer. PMID- 16349406 TI - Metabolism of Pyrene by the Basidiomycete Crinipellis stipitaria and Identification of Pyrenequinones and Their Hydroxylated Precursors in Strain JK375. AB - The metabolism of pyrene, a polycyclic aromatic hydrocarbon, by submerged cultures of the basidiomycete Crinipellis stipitaria was studied. After incubation for 68 h at 25 degrees C in a 20-liter fermentor with complex medium and 20 mg of pyrene per liter, five metabolites were detected. The compounds were isolated by preparative high-performance liquid chromatography on RP18 and DIOL gels. By UV, infrared, and H nuclear magnetic resonance spectroscopy and mass spectrometry, 1-hydroxypyrene, 1,6-dihydroxypyrene, 1,8-dihydroxypyrene, 1,6 pyrenequinone, and 1,8-pyrenequinone were identified. 1,6- and 1,8 dihydroxypyrene were obtained from fungal cultures for the first time. The formation of these metabolites was confirmed by investigations with [4,5,9,10 C]pyrene. PMID- 16349407 TI - Pyrene Metabolism in Crinipellis stipitaria: Identification of trans-4,5-Dihydro 4,5-Dihydroxypyrene and 1-Pyrenylsulfate in Strain JK364. AB - The isolation and identification of two novel metabolites in the fungal metabolism of pyrene are described. The plant-inhabiting basidiomycete Crinipellis stipitaria JK364 metabolized pyrene, a polycyclic aromatic hydrocarbon containing four rings, when grown in submerged cultures in a medium containing malt extract, glucose, and yeast extract. In experiments with [C] pyrene, after 7 days of incubation 40% of the labeled substrate was converted into organic solvent-extractable metabolites. Metabolites isolated from cultures grown with pyrene were identified as 1-pyrenylsulfate and trans-4,5-dihydro-4,5 dihydroxypyrene. 1-Hydroxypyrene, the precursor of 1-pyrenylsulfate, was also detected. 1-Pyrenylsulfate was isolated from mycelial extracts, whereas trans-4,5 dihydro-4,5-dihydroxypyrene was recovered from the culture filtrate. Identification of the compounds was based on their UV spectra, mass spectra, and nuclear magnetic resonance spectra. This is the first report on the detoxification of a polycyclic aromatic hydrocarbon by a plant-inhabiting basidiomycete. The occurrence of 1-pyrenylsulfate and trans-4,5-dihydro-4,5 dihydroxypyrene among fungal metabolites of pyrene is also new. PMID- 16349408 TI - Methods for assessment of biodegradability of plastic films in soil. AB - Traditional and novel techniques were tested and compared for their usefulness in evaluating biodegrad-ability claims made for newly formulated "degradable" plastic film products. Photosensitized polyethylene (PE), starch-PE, extensively plasticized polyvinyl chloride (PVC), and polypropylene (PP) films were incorporated into aerobic soil. Biodegradation was measured for 3 months under generally favorable conditions. Carbon dioxide evolution, residual weight recovery, and loss of tensile strength measurements were supplemented, for some films, by gas chromatographic measurements of plasticizer loss and gel permeation chromatographic (GPC) measurement of polymer molecular size distribution. Six- and 12-week sunlight exposures of photosensitized PE films resulted in extensive photochemical damage that failed to promote subsequent mineralization in soil. An 8% starch-PE film and the plasticized PVC film evolved significant amounts of CO(2) in biodegradation tests and lost residual weight and tensile strength, but GPC measurements demonstrated that all these changes were confined to the additives and the PE and PVC polymers were not degraded. Carbon dioxide evolution was found to be a useful screening tool for plastic film biodegradation, but for films with additives, polymer biodegradation needs to be confirmed by GPC. Photochemical cross-linking of polymer strands reduces solubility and may interfere with GPC measurements of polymer degradation. PMID- 16349409 TI - Microbially Mediated Formation of Benzonaphthothiophenes from Benzo[b]thiophenes. AB - Studies of the microbial metabolism of benzo[b]thiophene (molecular weight 134) by three Pseudomonas isolates showed the formation of benzothiophene sulfoxide, benzothiophene sulfone, and a sulfur-containing metabolite with a molecular weight of 234. Desulfurization of the high-molecular-weight product with nickel boride gave 1-phenylnaphthalene, indicating that the metabolite was benzo[b]naphtho[1,2-d]thiophene. Similarly, the isolates were capable of producing the analogous dimethyl-substituted benzonaphthothiophenes from methylbenzothiophenes that had the methyl substitution on the benzene ring. The formation of benzo[b] naphtho[1,2-d]thiophene was also observed when a petroleum degrading mixed culture was incubated with benzothiophene-supplemented Prudhoe Bay crude oil. Investigations into the mechanism of formation of these high molecular-weight compounds showed that they resulted from an abiotic, Diels-Alder type condensation of two molecules of the sulfoxide, which were microbially produced from the respective benzothiophene, with the subsequent loss of two atoms of hydrogen and oxygen and one atom of sulfur. The condensation products also formed from the sulfoxides of benzothiophene and methylbenzothiophenes when the sulfoxides were enzymatically synthesized by oxidation of the benzothiophene with horse heart cytochrome c and H(2)O(2). PMID- 16349410 TI - Spatial variability in biodegradation rates as evidenced by methane production from an aquifer. AB - Accurate predictions of carbon and energy cycling rates in the environment depend on sampling frequencies and on the spatial variability associated with biological activities. We examined the variability associated with anaerobic biodegradation rates at two sites in an alluvial sand aquifer polluted by municipal landfill leachate. In situ rates of methane production were measured for almost a year, using anaerobic wells installed at two sites. Methane production ranged from 0 to 560 mumol . m . day at one site (A), while a range of 0 to 120,000 mumol . m . day was measured at site B. The mean and standard deviations associated with methane production at site A were 17 and 57 mumol . m . day, respectively. The comparable summary statistics for site B were 2,000 and 9,900 mumol . m . day. The coefficients of variation at sites A and B were 340 and 490%, respectively. Despite these differences, the two sites had similar seasonal trends, with the maximal rate of methane production occurring in summer. However, the relative variability associated with the seasonal rates changed very little. Our results suggest that (i) two spatially distinct sites exist in the aquifer, (ii) methanogenesis is a highly variable process, (iii) the coefficient of variation varied little with the rate of methane production, and (iv) in situ anaerobic biodegradation rates are lognormally distributed. PMID- 16349411 TI - Induction of Carbon Starvation-Induced Proteins in Vibrio vulnificus. AB - By using two-dimensional polyacrylamide gel electrophoresis of pulse-labelled proteins, carbon starvation-induced (Sti) proteins produced by Vibrio vulnificus were identified. At least 34 proteins were induced over a 26-h period of carbon starvation. Although the total rate of protein synthesis over the 26-h starvation period suggests that there is a dramatic decrease in total protein synthesis within the first hour of starvation, at least 23 of the Sti proteins were induced within the first 20 min of carbon depletion. Six temporal classes of proteins could be identified, with class A(ii) encompassing the largest (21 proteins) group. All of the proteins in this group could be characterized by one of two patterns of protein synthesis. The addition of chloramphenicol at sequential times throughout starvation revealed that proteins required for starvation survival are made within the first 4 h of starvation. PMID- 16349412 TI - Occurrence and Role of Di-myo-Inositol-1,1'-Phosphate in Methanococcus igneus. AB - Methanococcus igneus, a hyperthermophilic marine methanogen (optimum growth temperature of 88 degrees C) with a 25-min doubling time, synthesizes an unusual inositol phosphodiester which is present at high intracellular concentrations along with l-alpha-glutamate and beta-glutamate. Identification of this compound as a dimeric inositol phosphodiester (di-myo-inositol-1,1'-phosphate) was provided by two-dimensional nuclear magnetic resonance methods. The intracellular levels of all three negatively charged solutes (l-alpha-glutamate, beta glutamate, and the inositol phosphodiester) increase with increasing levels of external NaCl, although the inositol compound shows much smaller increases with increasing NaCl levels than the glutamate isomers. The turnover of these solutes was examined by CO(2)-pulse-CO(2)-chase experiments. The results indicated that both the beta-glutamate and the inositol phosphodiester behaved as compatible solutes and were not efficiently metabolized by cells as was l-alpha-glutamate. At a fixed external NaCl concentration, lower ammonium levels increased the fraction of the inositol dimer present in extracts. The most pronounced changes in di-myo-inositol-1,1'-phosphate occurred as a function of cell growth temperature. While the organism grows over a relatively wide temperature range, the phosphodiester accumulated only when M. igneus was grown at temperatures of >/=80 degrees C. Thus, this unusual compound is a non-nitrogen-containing osmolyte preferentially synthesized at high growth temperatures. PMID- 16349413 TI - Quantification of frankia strains and other root-associated bacteria in pure cultures and in the rhizosphere of axenic seedlings by high-performance liquid chromatography-based muramic Acid assay. AB - Application of a high-performance liquid chromatography-based muramic acid assay with precolumn fluorescence derivatization to quantification of root-associated bacteria was studied both in pure cultures and in the rhizosphere of axenic Festuca rubra seedlings. Quantities of muramic acid from acid-hydrolyzed cells of Frankia strains, Streptomyces griseoviridis, Enterobacter agglomerans, Klebsiella pneumoniae, Pseudomonas sp., and Bacillus polymyxa were mostly proportional to the respective cell protein and carbon quantities, but in some strains, culture age and particularly sporulation affected these ratios considerably. The muramic acid/cell protein ratio was generally 2 to 4 times higher in strains of the two actinomycete genera, Frankia and Streptomyces, than in the rest of the strains. Quantification of Frankia strains, S. griseoviridis, E. agglomerans, and Pseudomonas sp. was also attempted from the rhizosphere of F. rubra seedlings which had been inoculated with pure cultured bacteria and incubated briefly. It was possible to quantify Frankia cells by use of the muramic acid assay from both the root and the growth medium, whereas cells of the rest of the bacterial genera could only be detected in the medium. The detection limit for muramic acid was about 10 ng/ml hydrolysis volume, and from the Festuca rhizosphere, 28 to 63% of the muramic acid in the Frankia inoculum was recovered. PMID- 16349414 TI - Production of 10-Ketostearic Acid from Oleic Acid by Flavobacterium sp. Strain DS5 (NRRL B-14859). AB - A microbial isolate, Flavobacterium sp. strain DS5, produces 10-ketostearic acid (10-KSA) from oleic acid in 85% yield. This is the first report on this type of reaction catalyzed by a Flavobacterium strain. The product was purified to give white, plate-like crystals melting at 79.2 degrees C. The optimum time, pH, and temperature for the production of 10-KSA are 36 h, 7.5, and 30 degrees C, respectively. A small amount of 10-hydroxystearic acid (10-HSA) (about 10% of the amount of the main product, 10-KSA) is also produced during the bioconversion. 10 KSA is not further metabolized by strain DS5 and accumulates in the medium. In contrast to growing cells, a resting-cell suspension of strain DS5 produces 10 HSA and 10-KSA in a ratio of 1:3. The crude cell extract obtained from ultrasonic disruption of the cells converts oleic acid mainly to 10-HSA (10-HSA/10-KSA ratio, 97:3). This result strongly suggested that oleic acid is converted to 10 KSA via 10-HSA. Enzymes catalyzing the hydration and secondary alcohol dehydrogenation are cell associated. Product 10-HSA from strain DS5 is 66% enantiomeric excess in the 10(R) form. The oleic acid conversion enzyme(s) reacts with unsaturated fatty acids in the order oleic acid > palmitoleic acid > linoleic acid > linolenic acid > gamma-linolenic acid > myristoleic acid. PMID- 16349415 TI - Production, Purification, and Properties of a Thermostable beta-Glucosidase from a Color Variant Strain of Aureobasidium pullulans. AB - A color variant strain of Aureobasidium pullulans (NRRL Y-12974) produced beta glucosidase activity when grown in liquid culture on a variety of carbon sources, such as cellobiose, xylose, arabinose, lactose, sucrose, maltose, glucose, xylitol, xylan, cellulose, starch, and pullulan. An extracellular beta glucosidase was purified 129-fold to homogeneity from the cell-free culture broth of the organism grown on corn bran. The purification protocol included ammonium sulfate treatment, CM Bio-Gel A agarose column chromatography, and gel filtrations on Bio-Gel A-0.5m and Sephacryl S-200. The beta-glucosidase was a glycoprotein with native molecular weight of 340,000 and was composed of two subunits with molecular weights of about 165,000. The enzyme displayed optimal activity at 75 degrees C and pH 4.5 and had a specific activity of 315 mumol . min . mg of protein under these conditions. The purified beta-glucosidase was active against p-nitrophenyl-beta-d-glucoside, cellobiose, cellotriose, cellotetraose, cellopentaose, cellohexaose, and celloheptaose, with K(m) values of 1.17, 1.00, 0.34, 0.36, 0.64, 0.68, and 1.65 mM, respectively. The enzyme activity was competitively inhibited by glucose (K(i) = 5.65 mM), while fructose, arabinose, galactose, mannose, and xylose (each at 56 mM) and sucrose and lactose (each at 29 mM) were not inhibitory. The enzyme did not require a metal ion for activity, and its activity was not affected by p-chloromercuribenzoate (0.2 mM), EDTA (10 mM), or dithiothreitol (10 mM). Ethanol (7.5%, vol/vol) stimulated the initial enzyme activity by 15%. Glucose production was enhanced by 7.9% when microcrystalline cellulose (2%, wt/vol) was treated for 48 h with a commercial cellulase preparation (5 U/ml) that was supplemented with the purified beta glucosidase (0.21 U/ml) from A. pullulans. PMID- 16349416 TI - Ribotyping of Erwinia chrysanthemi Strains in Relation to Their Pathogenic and Geographic Distribution. AB - 16S and 23S rRNAs from Escherichia coli were used to study the relationship among a representative collection of strains of Erwinia chrysanthemi differing in their original host and geographical origin. Phenetic analysis of restriction fragment length polymorphisms allowed the distribution of the studied strains into seven clusters. These clusters were similar to those obtained by cladistic methods and appeared to correlate well with the established pathovars and biovars but to a lesser extent with geographical distribution. Except for two groups of strains defined as tropical and temperate isolates (clusters 3 and 4, respectively), our clustering correlated well with botanical classifications of host plants. However, the rRNA groupings were shown to be more discriminative than biovar analysis. To assess the relationship between rRNA clusters and pathogenicity, 12 representative strains from different clusters were tested for pathogenicity on different plants. The two typical symptoms, maceration and wilting, were observed for these strains. The occurrence of the tobacco hypersensitivity reaction for a subset of these strains is discussed in light of recent results concerning the presence of an hrp gene. Considering symptom expression only, rather than the capacity for plant infection, strains from the same cluster were shown to induce similar symptoms in test plants. Thus, since host specificity is still quite controversial, rRNA patterns may constitute a useful tool in taxonomic and epidemiological studies of Erwinia chrysanthemi species. PMID- 16349417 TI - Survival, Growth, and Localization of Epiphytic Fitness Mutants of Pseudomonas syringae on Leaves. AB - Among 82 epiphytic fitness mutants of a Pseudomonas syringae pv. syringae strain that were characterized in a previous study, 4 mutants were particularly intolerant of the stresses associated with dry leaf surfaces. These four mutants each exhibited distinctive behaviors when inoculated onto and into plant leaves. For example, while none showed measurable growth on dry potato leaf surfaces, they grew to different population sizes in the intercellular spaces of bean leaves and on dry bean leaf surfaces, and one mutant appeared incapable of growth in both environments although it grew well on moist bean leaves. The presence of the parental strain did not influence the survival of the mutants immediately following exposure of leaves to dry, high-light incubation conditions, suggesting that the reduced survival of the mutants did not result from an inability to produce extracellular factors in planta. On moist bean leaves that were colonized by either a mutant or the wild type, the proportion of the total epiphytic population that was located in sites protected from a surface sterilant was smaller for the mutants than for the wild type, indicating that the mutants were reduced in their ability to locate, multiply in, and/or survive in such protected sites. This reduced ability was only one of possibly several factors contributing to the reduced epiphytic fitness of each mutant. Their reduced fitness was not specific to the host plant bean, since they also exhibited reduced fitness on the nonhost plant potato; the functions altered in these strains are thus of interest for their contribution to the general fitness of bacterial epiphytes. PMID- 16349418 TI - Comparison of the Behavior of Epiphytic Fitness Mutants of Pseudomonas syringae under Controlled and Field Conditions. AB - The epiphytic fitness of four Tn5 mutants of Pseudomonas syringae that exhibited reduced epiphytic fitness in the laboratory was evaluated under field conditions. The mutants differed more from the parental strain under field conditions than under laboratory conditions in their survival immediately following inoculation onto bean leaves and in the size of the epiphytic populations that they established, demonstrating that their fitness was reduced more under field conditions than in the laboratory. Under both conditions, the four mutants exhibited distinctive behaviors. One mutant exhibited particularly large population decreases and short half-lives following inoculation but grew epiphytically at near-wild-type rates, while the others exhibited reduced survival only in the warmest, driest conditions tested and grew epiphytically at reduced rates or, in the case of one mutant, not at all. The presence of the parental strain, B728a, did not influence the survival or growth of three of the mutants under field conditions; however, one mutant, an auxotroph, established larger populations in the presence of B728a than in its absence, possibly because of cross-feeding by B728a in planta. Experiments with B728a demonstrated that established epiphytic populations survived exposure of leaves to dry conditions better than newly inoculated cells did and that epiphytic survival was not dependent on the cell density in the inoculum. Three of the mutants behaved similarly to two nonpathogenic strains of P. syringae, suggesting that the mutants may be altered in traits that are missing or poorly expressed in naturally occurring nonpathogenic epiphytes. PMID- 16349419 TI - Construction of a Novel Polychlorinated Biphenyl-Degrading Bacterium: Utilization of 3,4'-Dichlorobiphenyl by Pseudomonas acidovorans M3GY. AB - Pseudomonas acidovorans M3GY is a recombinant bacterium with the novel capacity to utilize a biphenyl congener chlorinated on both rings, 3,4'-dichlorobiphenyl (3,4'-DCBP), as a sole carbon and energy source. Strain M3GY was constructed with a continuous amalgamated culture apparatus (L. Krockel and D. D. Focht, Appl. Environ. Microbiol. 53:2470-2475, 1987) with P. acidovorans CC1(19), a chloroacetate and biphenyl degrader, and Pseudomonas sp. strain CB15(1), a biphenyl and 3-chlorobenzoate degrader. Genetic and phenotypic data showed the recipient parental strain to be P. acidovorans CC1 and the donor parental strain to be Pseudomonas sp. strain CB15. In growth experiments with 3,4'-DCBP as a sole source of carbon, cultures of strain M3GY increased in absorbance from 0.07 to 0.39 in 29 days while reaching a protein concentration of 58 mug ml and 67% substrate dehalogenation. 4-Chlorobenzoate was identified from culture supernatants of strain M3GY by gas chromatography-infrared spectrometry-mass spectrometry; this would be consistent with the oxidation of the m-chlorinated ring through the standard biphenyl pathway. 4-Chlorobenzoate was converted to 4 chlorocatechol, which was metabolized through the meta-fission pathway. The construction of P. acidovorans M3GY, with the novel capability to utilize 3,4' DCBP, thus involves the complete use of meta-fission pathways for sequential rupture of the biphenyl and chlorobenzoate rings. PMID- 16349420 TI - Binding of Insecticidal Crystal Proteins of Bacillus thuringiensis to the Midgut Brush Border of the Cabbage Looper, Trichoplusia ni (Hubner) (Lepidoptera: Noctuidae), and Selection for Resistance to One of the Crystal Proteins. AB - The susceptibility of Trichoplusia ni larvae to several Bacillus thuringiensis insecticidal crystal proteins (ICPs) was tested. Neonatal larvae proved to be susceptible to solubilized trypsin-treated CryIA(a), CryIA(b), and CryIA(c) (50% lethal concentrations [LC(50)s], 570, 480, and 320 ng/cm, respectively) but showed little susceptibility to CryIB and CryID (LC(50)s, 5,640 and 2,530 ng/cm, respectively). The toxicity of ICPs was correlated to binding to the epithelial brush border of the midgut, as revealed by immunocytochemical staining with monoclonal antibodies. In vitro binding experiments with iodinated ICPs and brush border membrane vesicles indicated that CryIA(b) and CryIA(c) share the same high affinity binding site, whereas CryIA(a) binds to a different one. The affinities of CryIA(b) and CryIA(c) for the binding site were similar (K(d) = 3.6 and 4.7 nM, respectively), and the mean binding-site concentration was 0.71 pmol/mg of vesicle protein. Selection of a population with increasing concentrations of CryIA(b) produced 31-fold resistance in seven generations. The realized heritability (h) was 0.19. The increase of homozygosity (for resistance factors) as selection proceeded was reflected in the increase in the slopes of the dose mortality curves. Resistance was specific for CryIA(b) and did not extend to CryIA(a) or even to CryIA(c). This result was not predicted by the binding-site model, in which CryIA(b) and CryIA(c) bind to the same high-affinity binding site. This result may suggest a more complicated relationship between in vitro binding of ICPs to specific sites in the epithelial membrane of the midgut and the in vivo toxic effect. PMID- 16349421 TI - Comparison of Disulfide Contents and Solubility at Alkaline pH of Insecticidal and Noninsecticidal Bacillus thuringiensis Protein Crystals. AB - We compared two insecticidal and eight noninsecticidal soil isolates of Bacillus thuringiensis with regard to the solubility of their proteinaceous crystals at alkaline pH values. The protein disulfide contents of the insecticidal and noninsecticidal crystals were equivalent. However, six of the noninsecticidal crystals were soluble only at pH values of >/=12. This lack of solubility contributed to their lack of toxicity. One crystal type which was soluble only at pH >/=12 (strain SHP 1-12) did exhibit significant toxicity to tobacco hornworm larvae when the crystals were presolubilized. In contrast, freshly prepared crystals from the highly insecticidal strain HD-1 were solubilized at pH 9.5 to 10.5, but when these crystals were denatured, by either 8 M urea or autoclave temperatures, they became nontoxic and were soluble only at pH values of >/=12. These changes in toxicity and solubility occurred even though the denatured HD-1 crystals were morphologically indistinguishable from native crystals. Our data are consistent with the view that insecticidal crystals contain distorted, destabilized disulfide bonds which allow them to be solubilized at pH values (9.5 to 10.5) characteristic of lepidopteran and dipteran larval midguts. PMID- 16349422 TI - Characterization of a Protease from a Psychrotroph, Pseudomonas fluorescens 114. AB - A psychrotrophic bacterium isolated from river sediment was identified as Pseudomonas fluorescens 114. It grew at 0 degrees C and optimally at 20 degrees C. The bacterium produced a protease with a molecular weight of 47,000, which was stable in the pH range of 5 to 9 and worked optimally between pH 6.5 and 10. Activity was optimal at 35 degrees C and was lost immediately at 50 degrees C and after 5 min at 45 degrees C. At 0, 10, and 20 degrees C, 24, 38, and 57% of optimal activity were observed, respectively. PMID- 16349423 TI - Organic-solvent-tolerant bacterium which secretes organic-solvent-stable lipolytic enzyme. AB - A bacterial strain which could be grown in a medium containing organic solvents and which could secrete lipolytic enzyme was isolated. The stability of the lipolytic activity of the supernatant of the culture increased significantly in the presence of organic solvents such as toluene, cyclohexane, ethanol, and acetone. PMID- 16349424 TI - Adsorption of dissolved organic matter to the inorganic filter substrate and its implications for C uptake measurements. AB - Inorganic carbon uptake rates for glass fiber-filtered samples are higher than those for membrane-filtered samples because of adsorption of dissolved organic matter to the filter substrate. Experimentally derived values for adsorption onto filters were as follows (relative units): GF/F filter, 1, quartz filter, 1.1, GF/C filter, 0.6; GN-6 Gelman filter, 0.1; Nuclepore and Poretics filter, 0.0; Anodisc filter, 0.4 to 1.9. PMID- 16349425 TI - Cotransformation and Targeted Gene Inactivation in the Maize Anthracnose Fungus, Glomerella graminicola. AB - Cotransformation of Glomerella graminicola was achieved with the G. graminicola genes TUB1R1 (encoding a beta-tubulin which confers resistance to the fungicide benomyl) and PYR1 (encoding orotate phosphoribosyl transferase, which confers pyrimidine prototrophy). The cotransformation frequency was about 30% when selection was for pyrimidine prototrophy (Pyr) and 87% when selection was for benomyl-resistant (Bml) transformants. Southern blots confirmed that both transforming DNAs had integrated into the genomes of transformants which were expressing both Pyr and Bml phenotypes. A plasmid, p23, which contained a truncated 500-bp segment representing the central region of the PYR1 gene was constructed. The plasmid was introduced with pCG7, containing TUB1R1, into G. graminicola M1.001 (Pyr Bml), and Bml transformants were selected. The Bml transformants were screened on medium which did not contain uridine in order to identify Pyr mutants created by integration of p23 at the PYR1 locus. None of the primary transformants were Pyr, but 0.2% of uninucleate conidia collected from the pooled primary transformants gave rise to Pyr auxotrophs. Southern blots representing two of these Pyr mutants confirmed that they had the expected homologous integration of p23 at the PYR1 locus. This suggested that integration resulted in production of two nonfunctional copies of the gene, one lacking the 5' sequences and the other lacking the 3' sequences. This study demonstrates the feasibility of using cotransformation to perform targeted gene disruptions in G. graminicola. PMID- 16349426 TI - Paper mill effluent decolorization by fifty streptomyces strains. AB - Fifty actinomycete strains isolated from lignocellulosic substrates were examined for the ability to remove the color from a paper mill effluent obtained after semichemical alkaline pulping of wheat straw. Streptomyces sp. strains UAH 15, UAH 23, UAH 30, and UAH 51 were selected for their ability to decolorize the effluent in a liquid medium containing 1% (wt/vol) glycerol, 0.2% (wt/vol) ammonium sulfate, and 80% (vol/vol) effluent. The highest levels of decolorization achieved after the strains grew were 60 to 65%. Strains UAH 30 and UAH 51 were selected for further study because of their different patterns of effluent decolorization during growth. Fractionation of the decolorized effluent by gel permeation chromatography demonstrated that there were reductions in the levels of absorbance of the high- and medium-molecular-weight compounds. These fractions were mainly responsible for the color of the effluent, while the last fractions, the low-molecular-weight compounds, could have been responsible for the residual color of the decolorized effluent. Thin-layer chromatography revealed significant differences among the patterns of bands corresponding to the acidified supernatants obtained after precipitation of alkali-lignin from the effluent samples decolorized by different Streptomyces strains. PMID- 16349427 TI - Carbon Source Requirements for Exopolysaccharide Production by Lactobacillus casei CG11 and Partial Structure Analysis of the Polymer. AB - Exopolysaccharide production by Lactobacillus casei CG11 was studied in basal minimum medium containing various carbon sources (galactose, glucose, lactose, sucrose, maltose, melibiose) at concentrations of 2, 5, 10, and 20 g/liter. L. casei CG11 produced exopolysaccharides in basal minimum medium containing each of the sugars tested; lactose and galactose were the poorest carbon sources, and glucose was by far the most efficient carbon source. Sugar concentrations had a marked effect on polymer yield. Plasmid-cured Muc derivatives grew better in the presence of glucose and attained slightly higher populations than the wild-type strain. The values obtained with lactose were considerably lower for both growth and exopolysaccharide yield. The level of specific polymer production per cell obtained with glucose was distinctively lower for Muc derivatives than for the Muc strain. The polymer produced by L. casei CG11 in the presence of glucose was different from that formed in the presence of lactose. The polysaccharide produced by L. casei CG11 in basal minimum medium containing 20 g of glucose per liter had an intrinsic viscosity of 1.13 dl/g. It was rich in glucose (76%), which was present mostly as 2- or 3-linked residues along with some 2,3 doubly substituted glucose units, and in rhamnose (21%), which was present as 2-linked or terminal rhamnose; traces of mannose and galactose were also present. PMID- 16349428 TI - A structure-activity study with aryl acylamidases. AB - We examined the relationship between chemical structure and biodegradability of acylanilide herbicides by using a set of model compounds. Four bacterial isolates (one gram-negative and three gram-positive) that grew on acetanilide were used. These soil isolates cleaved the amide bond of acetanilide via an aryl acylamidase reaction, producing aniline and the organic acid acetate. A series of acetanilide analogs with alkyl substitutions on the nitrogen atom or the aromatic ring were tested for their ability to induce aryl acylamidase activity and act as substrates for the enzyme. The substrate range, in general, was limited to those analogs not disubstituted in the ortho position of the benzene ring or which did not contain an alkyl group on the nitrogen atom. These same N-substituted compounds did not induce enzyme activity either, whereas the ortho-substituted compounds could in some cases. PMID- 16349429 TI - Methane metabolism in a temperate swamp. AB - Comparisons between in situ CH(4) concentration and potential factors controlling its net production were made in a temperate swamp. Seasonal measurements of water table level and depth profiles of pH, dissolved CH(4), CO(2), O(2), SO(4), NO(3), formate, acetate, propionate, and butyrate were made at two adjacent sites 1.5 to 2 m apart. Dissolved CH(4) was inversely correlated to O(2) and, in general, to NO(3) and SO(4), potential inhibitors of methanogenesis. At low water table levels (August 1992), maximal CH(4) (2 to 4 muM) occurred below 30 cm, whereas at high water table levels (October 1992) or under flooded conditions (May 1993), CH(4) maxima (4 to 55 muM) occurred in the top 10 to 20 cm. Higher CH(4) concentrations were likely supported by inputs of fresh organic matter from decaying leaf litter, as suggested by high acetate and propionate concentrations (25 to 100 muM) in one of the sites in fall and spring. Measurements of potential CH(4) production (and consumption) showed that the highest rates generally occurred in the top 10 cm of soil. Soil slurry incubations confirmed the importance of organic matter to CH(4) production but also showed that competition for substrates by nonmethanogenic microorganisms could greatly attenuate its effect. PMID- 16349430 TI - Inoculation of Acacia mangium with Alginate Beads Containing Selected Bradyrhizobium Strains under Field Conditions: Long-Term Effect on Plant Growth and Persistence of the Introduced Strains in Soil. AB - The growth response of Acacia mangium Willd. to inoculation with selected Bradyrhizobium strains was investigated in two field trials in the Ivory Coast (West Africa). In the first trial (Anguededou), four provenances (i.e., trees originating from seeds harvested in different geographical areas) of A. mangium were inoculated with four Bradyrhizobium strains from different origins. Six months after being transplanted in the field, the heights of all inoculated trees showed a statistically significant increase of 9 to 26% compared with those of uninoculated trees, with the most effective strain being Aust 13c. After 19 months, the positive effect of inoculation on tree growth was confirmed. The effect of A. mangium provenance on tree growth was also highly significant. Trees from the Oriomo provenance of Papua New Guinea had a mean height that was 25% greater than those of other provenances. Analysis of variance showed a highly significant effect of interaction between strain and host provenance factors. Thus, most effective strain x provenance combinations could be proposed. Immunological identification of strains clearly showed that 90 to 100% of nodules from trees inoculated with three of the four Bradyrhizobium strains or from uninoculated trees contained exclusively Aust 13c 23 months after tree transplantation. This predominance of Aust 13c in nodules was still observed 42 months after tree transplantation. The second experiment (Port-Bouet), performed with a different soil, confirmed the long-term positive effect of Aust 13c on plant growth, its high competitive ability against indigenous strains, and its persistence in soil. Strain Aust 13c should thus be of great interest for inoculating A. mangium under a wide range of field conditions. PMID- 16349431 TI - Amino Acid Cycling in Colonies of the Planktonic Marine Cyanobacterium Trichodesmium thiebautii. AB - We examined diel trends in internal pools and net efflux of free amino acids in colonies of the nonheterocystous, diazotrophic cyanobacterium Trichodesmium thiebautii, freshly collected from waters of the Caribbean and the Bahamas. The kinetics of glutamate uptake by whole colonies were also examined. While intracolonial pools of most free amino acids were relatively constant through the day, pools of glutamate and glutamine varied over the diel cycle, with maxima during the early afternoon. This paralleled the daily cycle of nitrogenase activity. We also observed a large net release of these two amino acids from intact colonies. Glutamate release was typically 100 pmol of N colony h. This is about one-fourth the concurrent rate of N(2) fixation during the day. However, while nitrogenase activity only occurs during the day, net release of glutamate and glutamine persisted into the night and may therefore account for a greater loss of recently fixed N on a daily basis. This release may be an important route of new N input into tropical, oligotrophic waters. Whole colonies also displayed saturation kinetics with respect to glutamate uptake. The K(s) for whole colonies varied from 1.6 to 3.2 muM, or about 100-fold greater than typical ambient concentrations. Thus, uptake systems appear to be adapted to the higher concentrations of glutamate found within the intracellular spaces of the colonies. This suggests that glutamate may be a vehicle for N exchange among trichomes in the colony. PMID- 16349432 TI - Release of Dissolved Organic Nitrogen by Marine Diazotrophic Cyanobacteria, Trichodesmium spp. AB - Trichodesmium sp. is a filamentous, colonial cyanobacterium which contributes substantially to the input of nitrogen in tropical and subtropical oceanic waters through nitrogen fixation (N(2) fixation). We applied a N tracer technique to assess the rate of release of dissolved organic nitrogen (DON) from this cyanobacterium and compared those rates with rates of N(2) fixation determined for the same assemblages at the same times of day. Rates of release of DON showed considerable variation within replicate experiments and were variable depending on time of day and duration of time course experiments. On average, rates of DON release were ca. 50% the rates of N(2) fixation. We also fractionated the DON released by using ultrafiltration and found that 60 to 80% of the total organic release was of the size class <10,000 Da. The release of these organic compounds by Trichodesmium spp. is likely a significant source of new nitrogen for the associated bacteria or the non-nitrogen-fixing filaments of the Trichodesmium colonies. PMID- 16349433 TI - Investigation of an Iron-Oxidizing Microbial Mat Community Located near Aarhus, Denmark: Field Studies. AB - We investigated the microbial community that developed at an iron seep where anoxic groundwater containing up to 250 muM Fe flowed out of a rock wall and dense, mat-like aggregations of ferric hydroxides formed at the oxic-anoxic interface. In situ analysis with oxygen microelectrodes revealed that the oxygen concentrations in the mat were rarely more than 50% of air saturation and that the oxygen penetration depth was quite variable, ranging from <0.05 cm to several centimeters. The bulk pH of the mat ranged from 7.1 to 7.6. There appeared to be a correlation between the flow rates at different subsites of the mat and the morphotypes of the microorganisms and Fe oxides that developed. In subsites with low flow rates (<2 ml/s), the iron-encrusted sheaths of Leptothrix ochracea predominated. Miniature cores revealed that the top few millimeters of the mat consisted primarily of L. ochracea sheaths, only about 7% of which contained filaments of cells. Deeper in the mat, large particulate oxides developed, which were often heavily colonized by unicellular bacteria that were made visible by staining with acridine orange. Direct cell counts revealed that the number of bacteria increased from approximately 10 to 10 cells per cm and the total iron concentration increased from approximately 0.5 to 3 mmol/cm with depth in the mat. Primarily because of the growth of L. ochracea, the mat could accrete at rates of up to 3.1 mm/day at these subsites. The iron-encrusted stalks of Gallionella spp. prevailed in localized zones of the same low-flow-rate subsites, usually close to where the source water emanated from the wall. These latter zones had the lowest O(2) concentrations (<10% of the ambient concentration), confirming the microaerobic nature of Gallionella spp. In subsites with high flow rates (>6 ml/s) particulate Fe oxides were dominant; direct counts revealed that up to 10 cells of primarily unicellular bacteria per cm were associated with these particulate oxides. These zones exhibited little vertical stratification in either the number of cells or iron concentration. Finally, mat samples incubated anaerobically in the presence of acetate or succinate exhibited significant potential for iron reduction, suggesting the possibility that a localized iron cycle could occur within the mat community. PMID- 16349434 TI - Investigation of an Iron-Oxidizing Microbial Mat Community Located near Aarhus, Denmark: Laboratory Studies. AB - We constructed a small flow chamber in which suboxic medium containing 60 to 120 muM FeCl(2) flowed up through a sample well into an aerated reservoir, thereby creating an suboxic-oxic interface similar to the physicochemical conditions that exist in natural iron seeps. When microbial mat material from the Marselisborg iron seep that contained up to 10 bacterial cells per cm (D. Emerson and N. P. Revsbech, Appl. Environ. Microbiol. 60:4022-4031, 1994) was placed in the sample well of the chamber, essentially all of the Fe flowing through the sample well was oxidized at rates of up to 1,200 nmol of Fe oxidized per h per cm of mat material. The oxidation rates of samples of the mat that were pasteurized prior to inoculation were only about 20 to 50% of the oxidation rates of unpasteurized samples. Sodium azide also significantly inhibited oxidation. These results suggest that at least 50% and up to 80% of the Fe oxidation in the chamber were actively mediated by the microbes in the mat. It also appeared that Fe stimulated the growth of the community since chambers fed with FeCl(2) accumulated masses of either filamentous or particulate growth, both in the sample well and attached to the walls of the chamber. Control chambers that did not receive FeCl(2) showed no sign of such growth. Furthermore, after 4 to 5 days the chambers fed with FeCl(2) contained 35 to 75% more protein than chambers not supplemented with FeCl(2). Leptothrix ochracea and, to a lesser extent, Gallionella spp. were responsible for the filamentous growth, and the sheaths and stalks, respectively, of these two organisms harbored large numbers of Fe-encrusted, nonappendaged unicellular bacteria. In chambers where particulate growth predominated, the unicellular bacteria alone appeared to be the primary agents of iron oxidation. These results provide the first clear evidence that the "iron bacteria" commonly found associated with neutral-pH iron seeps are responsible for most of the iron oxidation and that the presence of ferrous iron appears to stimulate the growth of these organisms. PMID- 16349435 TI - Metabolic Pathways Leading to Mercury Methylation in Desulfovibrio desulfuricans LS. AB - The synthesis of methylmercury by Desulfovibrio desulfuricans LS was investigated on the basis of C incorporation from precursors and the measurement of relevant enzyme activities in cell extracts. The previously observed incorporation of C-3 from serine into methylmercury was confirmed by measurement of relatively high activities of serine hydroxymethyltransferase and other enzymes of this pathway. High rates of label incorporation into methylmercury from HCOO and HCO(3) prompted the assay of enzymes of the acetyl coenzyme A (CoA) synthase pathway. These enzymes were found to be present but at activity levels much lower than those reported for acetogens. Propyl iodide inhibited methylmercury and acetyl CoA syntheses to similar extents, and methylmercury synthesis was found to compete with acetyl-CoA synthesis for methyl groups. On the basis of these findings, we propose that in methylmercury synthesis by D. desulfuricans LS the methyl group is transferred from CH(3)-tetrahydrofolate via methylcobalamin. The methyl group may originate from C-3 of serine or from formate via the acetyl-CoA synthase pathway. These pathways are not unique to D. desulfuricans LS, and thus the ability of this bacterium to methylate mercury is most likely associated with the substrate specificity of its enzymes. PMID- 16349436 TI - Efficient Transformation of the Cephamycin C Producer Nocardia lactamdurans and Development of Shuttle and Promoter-Probe Cloning Vectors. AB - A high transformation efficiency (1 x 10 to 7 x 10 transformants per mug of DNA) of Nocardia lactamdurans LC411 was obtained by direct treatment of mycelium with polyethylene glycol 1000 and cesium chloride. A variety of vectors from Streptomyces lividans, Brevibacterium lactofermentum, Rhodococcus fascians, and a Nocardia (Amycolatopsis) sp. were tested; transformants could be obtained only with vectors derived from an endogenous plasmid of the Amycolatopsis sp. strain DSM 43387. Vectors carrying the kanamycin resistance gene (kan) as a selective marker were constructed. The transformation procedure has been optimized by using one of these vectors (pULVK1) and studying the influence of the age of the culture, concentrations of cesium chloride and polyethylene glycol, amount of plasmid DNA, and nutrient supplementations of the growth medium. Versatile shuttle cloning vectors (pULVK2 and pULVK3) have been developed by subcloning the pBluescript KS(+) multiple cloning site or a synthetic polylinker containing several unique restriction sites (EcoRV, DraI, BamHI, SstI, EcoRI, and HindIII). A second marker, the apramycin resistance gene (amr) has been added to the vectors (pULVK2A), allowing insertional inactivation of one of the markers while using the second one for selection. An alternative marker, the amy gene of Streptomyces griseus (pULAM2), which is easily detected by the release of extracellular amylase in transformants of N. lactamdurans carrying this vector, has been added. Two promoter-probe plasmids, pULVK4 and pULVK5, have been constructed, with the promoterless xylE gene as a reporter, for utilization in N. lactamdurans. PMID- 16349437 TI - Comparison of reactors for oxygen-sensitive reactions: reductive dechlorination of chlorophenols by vitamin b(12s). AB - Serum bottles are frequently used for studies of reductive dechlorination by vitamin B(12), but reducing conditions can be maintained only for several days. This time period is inadequate for evaluating the reductive dechlorination of some slow-reacting aromatic compounds. Sealed glass ampoules maintain reducing conditions for many months, but this method has the disadvantage of disallowing subsampling of the reaction mixture. A glass serum tube was modified for these experiments which not only maintained anoxic conditions for several days but also allowed subsamples to be removed during experiments. The modification was a restriction placed in the middle of the tube by heating in a flame, creating two chambers separated by a narrow neck. The lower chamber contained the oxygen sensitive reaction mixture. The upper chamber, sealed with a septum and screw cap, was purged with purified nitrogen or argon introduced and vented through fused silica capillaries. Reductive dechlorination of chlorophenols by vitamin B(12) reduced with Ti(III) citrate was monitored in all three reactor types. Sealed ampoules maintained reducing conditions for up to 12 months. The two chambered reactor maintained reducing conditions longer than the serum vials when frequent samples were taken. PMID- 16349438 TI - Regiospecificity of chlorophenol reductive dechlorination by vitamin b(12s). AB - Vitamin B(12), reduced by titanium (III) citrate to vitamin B(12s), catalyzes the reductive dechlorination of chlorophenols. Reductive dechlorination of pentachlorophenol and of all tetrachlorophenol and trichlorophenol isomers was observed. Reaction of various chlorophenols with vitamin B(12) favored reductive dechlorination at positions adjacent to another chlorinated carbon, but chlorines ortho to the hydroxyl group of a phenol were particularly resistant to reductive dechlorination, even if they were also ortho to a chlorine. This resulted in a reductive dechlorination pattern favoring removal of para and meta chlorines, which differs substantially from the pattern exhibited by anaerobic microbial consortia. PMID- 16349439 TI - Utilization of dissolved nitrogen by heterotrophic bacterioplankton: a comparison of three ecosystems. AB - The contributions of different organic and inorganic nitrogen and organic carbon sources to heterotrophic bacterioplankton in batch cultures of oceanic, estuarine, and eutrophic riverine environments were compared. The importance of the studied compounds was surprisingly similar among the three ecosystems. Dissolved combined amino acids (DCAA) were most significant, sustaining from 10 to 45% of the bacterial carbon demands and from 42 to 112% of the bacterial nitrogen demands. Dissolved free amino acids (DFAA) supplied 2 to 7% of the carbon and 6 to 24% of the nitrogen incorporated into the bacterial biomass, while dissolved DNA (D-DNA) sustained less than 5 and 12% of the carbon and nitrogen requirements, respectively. Ammonium was the second most important source of nitrogen, meeting from 13 to 45% of the bacterial demand in the oceanic and estuarine cultures and up to 270% of the demand in riverine cultures. Nitrate was taken up in the oceanic cultures (uptake equaled up to 46% of the nitrogen demand) but was released in the two others. Assimilation of DCAA, DFAA, and D-DNA combined supplied 43% of the carbon demand of the bacteria in the oceanic cultures, while approximately 25% of the carbon requirements were met by the three substrates at the two other sites. Assimilation of nitrogen from DCAA, DFAA, D-DNA, NH(4), and NO(3), on the other hand, exceeded production of particulate organic nitrogen in one culture at 27 h and in all cultures over the entire incubation period (50 h). These results suggest that the studied nutrient sources may fully support the nitrogen needs but only partially support the carbon needs of microbial communities of geographically different ecosystems. Furthermore, a comparison of the initial concentrations of the different substrates indicated that relative pool sizes of the substrates seemed to influence which substrates were primarily being utilized by the bacteria. PMID- 16349440 TI - Utilization of dissolved nitrogen by heterotrophic bacterioplankton: effect of substrate c/n ratio. AB - The significance of dissolved combined amino acids (DCAA), dissolved free amino acids (DFAA), and dissolved DNA (D-DNA) as sources of C and N for marine bacteria in batch cultures with variable substrate C/N ratios was studied. Glucose, ammonium, alanine, and phosphate were added to the cultures to produce C/N ratios of 5, 10, and 15 and to ensure that phosphorus was not limiting. Maximum bacterial particulate organic carbon production (after 25 h of incubation) was inversely correlated with the C/N ratio: with the addition of identical amounts of carbon, the levels of production were 9.0-, 10.0-, and 11.1-fold higher at C/N ratios of 15, 10, and 5, respectively, relative to an unamended control. The bacterial growth efficiency increased from 22% (control cultures) to 44 to 53% in the cultures with manipulated C/N ratios (C/N-manipulated cultures). Net carbon incorporation from DCAA, DFAA, and D-DNA supported on average 19, 4, and 3% (control cultures and cultures to which only phosphate was added [+P cultures]) and 5, 4, and 0.3% of the particulate organic carbon production (C/N-manipulated cultures), respectively. In the C/N-manipulated cultures, a 2.6- to 3.4-fold higher level of incorporation of DCAA, relative to that in the control cultures, occurred. Incorporation of D-DNA increased with the substrate C/N ratio, suggesting that D-DNA mainly was a source of N to the bacteria. Organic N (DCAA, DFAA, and D-DNA) sustained 14 to 49% of the net bacterial N production. NH(4) was the dominant N source and constituted 55 to 99% of the total N uptake. NO(3) contributed up to 23% to the total N uptake but was released in two cultures. The studied N compounds sustained nearly all of the bacterial N demand. Our results show that the C/N ratio of dissolved organic matter available to bacteria has a significant influence on the incorporation of individual compounds like DCAA and D-DNA. PMID- 16349441 TI - Production and Characterization of an Exopolysaccharide Excreted by a Deep-Sea Hydrothermal Vent Bacterium Isolated from the Polychaete Annelid Alvinella pompejana. AB - The heterotrophic and mesophilic marine bacterium HYD-1545 was isolated on a metal-amended medium from the dorsal integument of the hydrothermal vent polychaete Alvinella pompejana. This strain, which can be assigned to the genus Alteromonas on the basis of its G+C content and phenotypical features, produced large amounts of an acidic polysaccharide in batch cultures. The polysaccharide was excreted during the stationary phase of growth and contained glucose, galactose, glucuronic acid, galacturonic acid, and 4,6-O-(1-carboxyethilidene) galactose as major components. This polysaccharide was a polyelectrolyte, and the viscosity of its solutions depended on the ionic strength. The decrease in viscosity with increasing NaCl concentrations and the effect of Ca in decreasing the viscosity at low Ca concentrations support a model in which the polysaccharide carries anionic groups. However, an unusual behavior was observed at higher concentrations and could be related to intermolecular interactions involving Ca ions. PMID- 16349442 TI - Hydrolysis of Oleuropein by Lactobacillus plantarum Strains Associated with Olive Fermentation. AB - Oleuropein (Chemical Abstracts Service registry number 32619-42-4), a bitter tasting secoiridoid glucoside commonly found in leaves of the olive tree as well as in olives (Olea europaea L.), was found to be hydrolyzed by the beta glucosidase (EC 3.2.1.2.1) produced by oleuropeinolytic Lactobacillus plantarum type strains. Three strains, designated B17, B20, and B21, were isolated from the brine of naturally ripe olives not treated with alkali. These strains were rod shaped forms, grown at a pH 3.5 limit, and tolerated 1% oleuropein and 8% NaCl in the growth medium. The beta-glucosidase produced hydrolyzed 5-bromo-4-chloro-3 indolyl-beta-d-glucopy-ranoside as well as oleuropein. The presence of 2% glucose in the medium inhibited activity by 40 to 50%, depending on the bacterial strain. Chromatographic analysis of the trimethylsilyl derivatives of the products obtained after 7 days of incubation at 30 degrees C of strain B21 showed all the hydrolysis products of oleuropein, i.e., aglycone, iridoid monoterpen, and 3,4 dihydroxyphenylethanol (hydroxytyrosol). Oleuropein and its aglycone after 21 days of incubation decreased to trace levels with the simultaneous increase in concentration of beta-3,4-dihydroxyphenylethanol. PMID- 16349443 TI - A simple and widely applicable method for preparing homogeneous and stable quality control samples in water microbiology. AB - Test strains suspended in skim milk, quickly frozen in dry ice-ethanol, and stored at - 70 degrees C can be used as quality control samples that are immediately available by quickly thawing at 37 degrees C. The samples remain homogeneous and stable for at least 1 year, except for Aeromonas hydrophila, which decreases 20 to 30% in 1 year. PMID- 16349444 TI - Formaldehyde Solution Effectively Inactivates Spores of Bacillus anthracis on the Scottish Island of Gruinard. AB - Gruinard Island was heavily contaminated with the spores of virulent Bacillus anthracis during biological weapons trials in World War II. However, an extensive survey in 1979 showed that most of the island was not contaminated. In the early 1980s, a more intensive survey revealed that the contamination was largely confined to the top 8 cm of the soil in a 2.6-ha area of the 211-ha island. Small scale tests showed that the spores could be inactivated by drenching the soil with fluid biocides. A solution of 5% formaldehyde in seawater applied by surface spray to each square meter of ground was shown to be the most effective treatment and was utilized for large-scale decontamination of the affected areas. Following this treatment, extensive sampling revealed that most of the spores of B. anthracis had been inactivated. Isolated pockets of surviving spores were treated further. A flock of sheep was then allowed to graze over the entire island for 5 months; none contracted anthrax. PMID- 16349445 TI - Hydrostatic pressure and electroporation have increased bactericidal efficiency in combination with bacteriocins. AB - We report here that both ultrahigh hydrostatic pressure (UHP) and electroporation (EP) induced sublethal injury to bacterial cells surviving the treatments. The injured cells of both gram-positive and -negative bacteria became sensitive to the bacteriocins pediocin AcH and nisin. Bacteriocins in combination with either UHP or EP have greater antibacterial effectiveness than UHP or EP alone. PMID- 16349446 TI - New composite biocarriers engineered to contain adsorptive and ion-exchange properties improve immobilized-cell bioreactor process dependability. AB - Novel biocarriers that combine the adsorptive properties of activated carbon with the ion-exchange properties of zeolite-based type Z inorganic oxide biocarriers (D. R. Durham, L. C. Marshall, J. G. Miller, and A. B. Chmurny, Appl. Environ, Microbiol. 60:3329-3335, 1994.) were developed. These biocarriers, designated Type CZ, possess fundamental properties that heretofore have not been described for available microbial immobilization matrices. Type CZ biocarriers provide an environment that promotes dense microbial colonization and maintains bioreactor productivity by buffering immobilized microorganisms from unfavorable operating conditions. Data demonstrating protection of immobilized bacteria from organic shock loads and extended pH shocks are presented. In addition, bioreactors containing the composite Type CZ biocarriers continue to remove waste stream contaminants during periods of oxygen deprivation and nutrient limitation. PMID- 16349447 TI - Reversible Conversion of 4-Hydroxybenzoate and Phenol by Clostridium hydroxybenzoicum. AB - Reversible conversion of 4-hydroxybenzoate and phenol and their analogs was observed in whole-cell suspensions and cell extracts of Clostridium hydroxybenzoicum grown with 4-hydroxybenzoate and 3,4-dihydroxybenzoate. Assuming that bicarbonate is the cosubstrate, the equilibrium constants calculated for the reactions 4-hydroxybenzoate + H(2)O left arrow over right arrow phenol + HCO(3) and 3,4-dihydroxybenzoate + H(2)O left arrow over right arrow catechol + HCO(3) were 11.4 (+/- 0.5) and 5.05 (+/- 0.25), respectively. In a phenol-adapted sediment slurry, 4-hydroxybenzoate and 3,4-dihydroxybenzoate were decarboxylated to phenol and to catechol, respectively, as intermediates without a lag time. PMID- 16349448 TI - Efficacy of Nucleic Acid Probes for Detection of Poliovirus in Water Disinfected by Chlorine, Chlorine Dioxide, Ozone, and UV Radiation. AB - MilliQ water was inoculated with poliovirus type 1 strain LSc-1 and was treated with disinfectants, including chlorine, chlorine dioxide, ozone, and UV light. No relationship between probes and plaque assays were seen, demonstrating that viral nucleic acids were not destroyed. These findings suggest that nucleic acid probes cannot distinguish between infectious and noninfectious viruses and cannot be used in the evaluation of treated waters. PMID- 16349449 TI - Microbial Production of Poly-beta-Hydroxybutyric Acid from d-Xylose and Lactose by Pseudomonas cepacia. AB - Poly-beta-hydroxybutyric acid (PHB) was produced from xylose and lactose by using Pseudomonas cepacia. Approximately 50% PHB (grams of PHB total/grams of biomass total) was produced. With a laser-based fluorescent probe, beta-galactosidase activity was shown to be induced in P. cepacia cells grown on lactose but not in those grown on glucose or xylose. P. cepacia has the potential to produce biodegradable thermoplastics from hemicellulosic hydrolysates and cheese whey. PMID- 16349450 TI - New method for isolating barophiles from intestinal contents of deep-sea fishes retrieved from the abyssal zone. AB - We devised a new method (the dorayaki method) using marine agar under in situ pressures to isolate barophilic bacteria from the intestinal contents of three deep-sea fishes (two Coryphaenoides yaquinae samples and one Ilyophis sp. sample) retrieved from depths of 4,700 to 6,100 m in the Northwest Pacific Ocean. All 10 strains isolated from one sample (C. yaquinae) were obligately barophilic. One of the 10 strains did not grow at atmospheric pressure and 103.4 MPa but did grow well between 20.7 and 82.7 MPa, with optimal growth at 41.4 MPa. This method is useful for isolating psychrophilic and barophilic deep-sea bacteria. PMID- 16349451 TI - Alteration of Substrate Specificity and Optimum pH of Sarcosine Oxidase by Random and Site-Directed Mutagenesis. AB - The sarcosine oxidase gene was randomly mutated, and F103V, which showed altered specificity, was screened with the plate assay. Five different mutants at position 103 were also constructed. Each mutant exhibited a rather striking change in substrate specificity, except for F103W. The optimum pH was also shifted, depending on the substrate. PMID- 16349452 TI - Effect of genetically modifying the lactococcal proteolytic system on ripening and flavor development in cheddar cheese. AB - THREE BATCHES OF SIX CHEDDAR CHEESES WERE MANUFACTURED BY USING THE FOLLOWING LACTOCOCCAL STRAINS: (i) UC317 as a control; (ii) JL3601, a proteinase-negative derivative of UC317 transformed with high-copy-number plasmid pCI3601 containing the cloned proteinase gene complex from UC317; (iii) AM312, a proteinase-negative derivative of UC317 transformed with plasmid pMG36enpr containing the neutral proteinase gene from Bacillus subtilis; (iv) AC322, JL3601 transformed with pMG36enpr; (v) AC311, UC317 transformed with plasmid pNZ1120, which contains the aminopeptidase N (pepN) gene from Lactococcus lactis subsp. lactis MG1363; and (vi) AC321, JL3601 transformed with pNZ1120. Organoleptic and chemical analyses indicated that (i) the control cheeses, which were made with UC317, were of the highest quality; (ii) cheeses made with strains harboring pCI3601 in addition to either pMG36enpr (AC322) or pNZ1120 (AC321) did not ripen in a significantly different manner than cheeses made with AM312 (containing only pMG36enpr) or AC311 (containing only pNZ1120), respectively; (iii) cheeses made with strains that overproduce pepN did not have improved body, texture, and flavor characteristics; and (iv) cheeses made with strains harboring the neutral proteinase from B. subtilis (AM312 and AC322) underwent greatly accelerated proteolysis. PMID- 16349453 TI - Transfer in Marine Sediments of the Naturally Occurring Plasmid pRAS1 Encoding Multiple Antibiotic Resistance. AB - The results of microcosm experiments performed with the fish-pathogenic bacterium Aeromonas salmonicida acting as a donor showed that promiscuous plasmid pRAS1, which encodes tetracycline resistance, is transferred at a high frequency in marine sediments even in the absence of a selective factor. The presence of oxytetracycline resulted in an increase in the transfer frequency compared with that of a microcosm to which no selective factor was added. Transfer frequencies of 3.4 x 10 transconjugant per recipient and 3.6 transconjugants per donor cell were obtained in a microcosm to which oxytetracycline had been added. Hybridization with a DNA probe specific for plasmid pRAS1 revealed that 45.8% of the oxytetracycline-resistant isolates obtained from a microcosm with no selective pressure carried the plasmid, while 86.8% of the isolates obtained from a microcosm to which oxytetracycline had been added carried the plasmid. Phenotypic characterization of the transconjugants revealed that the plasmid had been transferred to a variety of different biotypes in both microcosms. The diversity among the transconjugants isolated from the microcosm to which oxytetracycline had been added was substantially lower than the diversity among the transconjugants isolated from the microcosm to which no selective agent had been added. PMID- 16349454 TI - Mimosine, a Toxin Present in Leguminous Trees (Leucaena spp.), Induces a Mimosine Degrading Enzyme Activity in Some Rhizobium Strains. AB - Thirty-seven Rhizobium isolates obtained from the nodules of leguminous trees (Leucaena spp.) were selected on the basis of their ability to catabolize mimosine, a toxin found in large quantities in the seeds, foliage, and roots of plants of the genera Leucaena and Mimosa. A new medium containing mimosine as the sole source of carbon and nitrogen was used for selection. The enzymes of the mimosine catabolic pathway were inducible and were present in the soluble fraction of the cell extract of induced cells. On the basis of a comparison of the growth rates of Rhizobium strains on general carbon and nitrogen sources versus mimosine, the toxin appears to be converted mostly to biomass and carbon dioxide. Most isolates able to grow on mimosine as a source of carbon and nitrogen are also able to utilize 3-hydroxy-4-pyridone, a toxic intermediate of mimosine degradation in other organisms. PMID- 16349455 TI - Induction of Chitin-Binding Proteins during the Specific Attachment of the Marine Bacterium Vibrio harveyi to Chitin. AB - Previous work has shown that attachment of Vibrio harveyi to chitin is specific and involves at least two chitin-binding peptides. However, the roles and regulation of these chitin-binding peptides in attachment are still unclear. Here we show that preincubation with the oligomeric sugars composing chitin stimulated chitinase activity, cellular attachment to chitin, and production of chitin binding peptides. One of these peptides, a 53-kDa peptide, is produced constitutively and appears to mediate initial attachment to chitin. Synthesis of another peptide, a 150-kDa chitin-binding peptide, is induced by chitin and thus may be involved in time-dependent attachment. Coordinated regulation of attachment and degradation of chitin may give bacteria like V. harveyi a selective advantage over other bacteria in nutrient-poor aquatic environments. PMID- 16349456 TI - Degradation of Raw Starch by a Wild Amylolytic Strain of Lactobacillus plantarum. AB - Lactobacillus plantarum A6, isolated from fermented cassava, can break down cassava raw starch that has not been subjected to preliminary physicochemical treatment. When the pH was kept at 6, the microorganism cultured in a bioreactor excreted a high alpha-amylase activity (60 U/ml). Synthesis of the enzyme occurred during the stationary phase and resulted in full hydrolysis of the cassava starch granules. This gave 41 g of lactic acid from 45 g of raw starch after 3 days of fermentation. Enzymatic attack was evident under scanning electron microscopy in the rougher appearance of the surface of starch granules and in the presence of large cavities in some of them. In contrast, when the pH was not regulated, only a small amount of alpha-amylase activity was produced (2 U/ml) and no decrease in the starch content of the medium was observed. However, under scanning electron microscopy, some granules displayed a rougher surface, which might have been the result of weak enzymatic attack. PMID- 16349457 TI - Involvement of an N-Acetylglucosaminidase in Autolysis of Propionibacterium freudenreichii CNRZ 725. AB - Propionibacterium freudenreichii plays an important role in Swiss cheese ripening (it produces propionic acid, acetic acid, and CO(2)). Moreover, autolysis of this organism certainly contributes to proteolysis and lipolysis of the curd because intracellular enzymes are released. By varying external factors, we determined the following conditions which promoted autolysis of both whole cells and isolated cell walls of P. freudenreichii CNRZ 725: (i) 0.1 M potassium phosphate buffer (pH 5.8) at 40 degrees C and (ii) 0.05 to 0.1 M KCl at 40 degrees C. We found that early-exponential-phase cells possessed the highest autolytic activity. It should be emphasized that the pH of Swiss cheese curd (pH 5.5 to 5.7) is near the optimal pH which we determined. Ultrastructural observations by electron microscopy revealed a 16-nm-thick homogeneous cell wall, as well as degradation of the cell wall that occurred concomitantly with cell autolysis. In the presence of 0.05 M potassium chloride, there was a great deal of isolated cell wall autolysis (the optical density at 650 nm decreased 77.5% +/- 7.3% in 3 h), and one-half of the peptidoglycan material was released. Finally, the main autolytic activity was due to an N-acetylglucosaminidase activity. PMID- 16349458 TI - Bleaching of Hardwood Kraft Pulp with Manganese Peroxidase Secreted from Phanerochaete sordida YK-624. AB - In vitro bleaching of an unbleached hardwood kraft pulp was performed with manganese peroxidase (MnP) from the fungus Phanerochaete sordida YK-624. When the kraft pulp was treated with partially purified MnP in the presence of MnSO(4), Tween 80, and sodium malonate with continuous addition of H(2)O(2) at 37 degrees C for 24 h, the pulp brightness increased by about 10 points and the kappa number decreased by about 6 points compared with untreated pulp. The pulp brightness was also increased by 43 points to 75.5% by multiple (six) treatments with MnP combined with alkaline extraction. Our results indicate that in vitro degradation of residual lignin in hardwood kraft pulp with MnP is possible. PMID- 16349460 TI - Digestion of herring by indigenous bacteria in the minke whale forestomach. AB - Northeastern Atlantic minke whales (Balaenoptera acutorostrata) have a multichambered stomach system which includes a nonglandular forestomach resembling that of ruminants. Bacteria from the forestomachs of herring-eating whales were enumerated and isolated in an anaerobic rumen-like culture medium (M8W medium). The total viable population of anaerobic bacteria ranged from 73 x 10 to 145 x 10/ml of forestomach fluid (n = 4). Lactobacillus spp. (19.7%), Streptococcus spp. (35.9%), and Ruminococcus spp. (12.8%) were the most common of the bacterial strains (n = 117) isolated by use of M8W medium from the forestomach fluid population of two minke whales. Most of the isolates stained gram positive (93.2%), 62.4% were cocci, and all strains were strictly anaerobic. The population of lipolytic bacteria in one animal, enumerated by use of a selective lipid medium, constituted 89.7% of the viable population. The total viable population of anaerobic bacteria in freshly caught and homogenized herring (Clupea harengus) ranged from 56.7 to 95.0 cells per gram of homogenized prey (n = 3) when M8W medium was used. Pediococcus spp. (30.6%) and Aerococcus spp. (25.0%) were most common of the bacterial strains (n = 72) isolated from the homogenized herring. Most of the bacterial strains were gram positive (80.6%), and 70.8% were cocci. Unlike the forestomach bacterial population, as many as 61.1% of the strains from the herring were facultatively anaerobic. All bacterial strains isolated from the prey had phenotypic patterns different from those of strains isolated from the dominant bacterial population in the forestomach, indicating that the forestomach microbiota is indigenous. Scanning electron microscopic examinations revealed large numbers of bacteria, surrounded by a glycocalyx, attached to partly digested food particles in the forestomach. These data support the hypothesis that symbiotic microbial digestion occurs in the forestomach and that the bacteria are indigenous to minke whales. PMID- 16349459 TI - Isolation of Insertion Sequence ISRLdTAL1145-1 from a Rhizobium sp. (Leucaena diversifolia) and Distribution of Homologous Sequences Identifying Cross Inoculation Group Relationships. AB - Insertion sequence (IS) element ISRLdTAL1145-1 from Rhizobium sp. (Leucaena diversifolia) strain TAL 1145 was entrapped in the sacB gene of the positive selection vector pUCD800 by insertional inactivation. A hybridization probe prepared from the whole 2.5-kb element was used to determine the distribution of homologous sequences in a diverse collection of 135 Rhizobium and Bradyrhizobium strains. The IS probe hybridized strongly to Southern blots of genomic DNAs from 10 rhizobial strains that nodulate both Phaseolus vulgaris (beans) and Leucaena leucocephala (leguminous trees), 1 Rhizobium sp. that nodulates Leucaena spp., 9 R. meliloti (alfalfa) strains, 4 Rhizobium spp. that nodulate Sophora chrysophylla (leguminous trees), and 1 nonnodulating bacterium associated with the nodules of Pithecellobium dulce from the Leucaena cross-inoculation group, producing distinguishing IS patterns for each strain. Hybridization analysis revealed that ISRLdTAL1145-1 was strongly homologous with and closely related to a previously isolated element, ISRm USDA1024-1 from R. meliloti, while restriction enzyme analysis found structural similarities and differences between the two IS homologs. Two internal segments of these IS elements were used to construct hybridization probes of 1.2 kb and 380 bp that delineate a structural similarity and a difference, respectively, of the two IS homologs. The internal segment probes were used to analyze the structures of homologous IS elements in other strains. Five types of structural variation in homolog IS elements were found. The predominate IS structural type naturally occurring in a strain can reasonably identify the strain's cross-inoculation group relationships. Three IS structural types were found in Rhizobium species that nodulate beans and Leucaena species, one of which included the designated type IIB strain of R. tropici (CIAT 899). Weak homology to the whole IS probe, but not with the internal segments, was found with two Bradyrhizobium japonicum strains. The taxonomic and ecological implications of the distribution of ISRLdTAL1145-1 are discussed. PMID- 16349462 TI - Coexistence among Epiphytic Bacterial Populations Mediated through Nutritional Resource Partitioning. AB - The levels of coexistence between Pseudomonas syringae and various nonpathogenic epiphytic species in the phyllosphere of beans (Phaseolus vulgaris) were assessed by using replacement series. The epiphytic species Pseudomonas fluorescens, Pantoea agglomerans, Stenotrophomonas maltophilia, and Methylobacterium organophilum were all capable of exhibiting higher levels of coexistence with P. syringae than was observed with a near-isogenic P. syringae strain pair. The ecological similarity of the epiphytes was estimated with niche overlap indices derived from in vitro carbon source utilization profiles. The level of coexistence of the epiphytes was inversely correlated with the ecological similarity of the strains. Hence, the level of coexistence between the epiphytes was proportional to the degree of niche differentiation, defined as the ability to utilize carbon sources not utilized by a competing strain. Comparisons of utilization profiles for groups of carbon sources (amino acids, organic acids, and carbohydrates) indicated the types of carbon sources for which the strains likely competed in the bean phyllosphere. P. fluorescens and P. syringae strains probably competed for most carbon sources. S. maltophilia and M. organophilum strains probably competed with P. syringae for most organic acids but few amino acids or carbohydrates. P. agglomerans strains probably competed with P. syringae for most amino acids and organic acids but few carbohydrates. A variable level of coexistence observed between P. agglomerans and P. syringae probably reflected the variability in abundance in the bean phyllosphere of the carbohydrates that P. agglomerans utilized exclusively. PMID- 16349461 TI - Uniparental Mitochondrial Transmission in the Cultivated Button Mushroom, Agaricus bisporus. AB - A uniparental mitochondrial (mt) transmission pattern has been previously observed in laboratory matings of the cultivated mushroom Agaricus bisporus on petri dishes. In this study, four sets of specific matings were further examined by taking mycelial plugs from the confluent zone of mated homokaryons and inoculating these plugs into rye grain for laboratory fruiting and for fruiting under industrial conditions. Examination of the mt genotype of each individual fruit body for mt-specific restriction fragment length polymorphisms further confirmed that the mt genome was inherited uniparentally. The vegetative radial growth and the fruiting activity of two pairs of intraspecific heterokaryons, each pair carrying the same combination of nuclear genomes but different mt genotypes, were compared. Our results suggested that the mt genotype did not appreciably affect radial growth or fruiting activity. The failure to recover both heterokaryons, each carrying either parental mt genotype in any given cross, therefore clearly indicated that in matings of A. bisporus, the mt genome from one of the parental homokaryons is either selectively excluded in the newly formed heterokaryon or selectively eliminated in the immediate heterokaryotic mitotic progeny of the newly formed heterokaryon. PMID- 16349463 TI - Pathological and Molecular Characterization of Xanthomonas campestris Strains Causing Diseases of Cassava (Manihot esculenta). AB - Fifty-one strains representing Xanthomonas campestris pv. manihotis and cassavae and different pathovars occurring on plants of the family Euphorbiaceae were characterized by ribotyping with a 16S+23S rRNA probe of Escherichia coli and by restriction fragment length polymorphism analysis with a plasmid probe from X. campestris pv. manihotis. Pathogenicity tests were performed on cassava (Manihot esculenta). Histological comparative studies were conducted on strains of two pathovars of X. campestris (vascular and mesophyllic) that attack cassava. Our results indicated that X. campestris pv. manihotis and cassavae have different modes of action in the host and supplemented the taxonomic data on restriction fragment length polymorphism that clearly separate the two pathovars. The plasmid probe could detect multiple restriction fragment length polymorphisms among strains of the pathovar studied. Ribotyping provides a useful tool for rapid identification of X. campestris pathovars on cassava. PMID- 16349464 TI - Production and Purification of Extracellular D-Xylose Isomerase from an Alkaliphilic, Thermophilic Bacillus sp. AB - An alkaliphilic, thermophilic Bacillus sp. (NCIM 59) produced extracellular xylose isomerase at pH 10 and 50 degrees C by using xylose or wheat bran as the carbon source. The distribution of xylose isomerase as a function of growth in comparison with distributions of extra- and intracellular marker enzymes such as xylanase and beta-galactosidase revealed that xylose isomerase was truly secreted as an extracellular enzyme and was not released because of sporulation or lysis. The enzyme was purified to homogeneity by ammonium sulfate precipitation followed by gel filtration, preparative polyacrylamide gel electrophoresis, and ion exchange chromatography. The molecular weight of xylose isomerase was estimated to be 160,000 by gel filtration and 50,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating the presence of three subunits. The enzyme is most active at pH 8.0 and with incubation at 85 degrees C for 20 min. Divalent metal ions Mg, Co, and Mn were required for maximum activity of the enzyme. The K(m) values for D-xylose and D-glucose at 80 degrees C and pH 7.5 were 6.66 and 142 mM, respectively, while K(cat) values were 2.3 x 10 s and 0.5 x 10 s, respectively. PMID- 16349465 TI - Effect of mineral and organic soil constituents on microbial mineralization of organic compounds in a natural soil. AB - This research addressed the effect of mineral and organic soil constituents on the fate of organic compounds in soils. Specifically, it sought to determine how the associations between organic chemicals and different soil constituents affect their subsequent biodegradation in soil. Four C-labeled surfactants were aseptically adsorbed to montmorillonite, kaolinite, illite, sand, and humic acids. These complexes were mixed with a woodlot soil, and CO(2) production was measured over time. The mineralization data were fitted to various production models by nonlinear regression, and a mixed (3/2)-order model was found to most accurately describe the mineralization patterns. Different mineralization patterns were observed as a function of the chemical and soil constituents. Surfactants that had been preadsorbed to sand or kaolinite usually showed similar mineralization kinetics to the control treatments, in which the surfactants were added to the soil as an aqueous solution. Surfactants that had been bound to illite or montmorillonite were typically degraded to lesser extents than the other forms, while surfactant-humic acid complexes were degraded more slowly than the other forms. The desorption coefficients (K(d)) of the soil constituent-bound surfactants were negatively correlated with the initial rates of degradation (k(1)) and estimates of CO(2) yield (P(o)) as well as actual total yields of CO(2). However, there was no relationship between K(d) and second-stage zero order rates of mineralization (k(o)). Microbial community characteristics (biomass and activity) were not correlated with any of the mineralization kinetic parameters. Overall, this study showed that environmental form had a profound effect on the ultimate fate of biodegradable chemicals in soil. This form is defined by the physicochemical characteristics of the chemical, the composition and mineralogy of the soil, and the mode of entry of the chemical into the soil environment. PMID- 16349466 TI - Precipitation of Metallic Cations by the Acidic Exopolysaccharides from Bradyrhizobium japonicum and Bradyrhizobium (Chamaecytisus) Strain BGA-1. AB - The interaction between the acidic exopolysaccharides produced by two Bradyrhizobium strains and several metal cations has been studied. Aqueous solutions in the millimolar range of Fe but not of Fe precipitated the exopolysaccharides from Bradyrhizobium (Chamaecytisus) strain BGA-1 and, to a lesser extent, Bradyrhizobium japonicum USDA 110. The precipitation was pH dependent, with a maximum around pH 3. The precipitate was redissolved by changing the pH and by Fe reduction or chelation. Deacetylation of B. japonicum polysaccharide increased its precipitation by Fe. At pH near neutrality, the polysaccharide from Bradyrhizobium (Chamaecytisus) strain BGA-1 stabilized Fe solutions, despite the insolubility of Fe(OH)(3). Aluminum precipitated Bradyrhizobium (Chamaecytisus) polysaccharide but not the polysaccharide produced by B. japonicum. The precipitation showed a maximum at about pH 4.8, and the precipitate was redissolved after Al chelation with EDTA. Precipitation was inhibited by increases in the ionic strength over 10 mM. Bradyrhizobium (Chamaecytisus) polysaccharide was also precipitated by Th, Sn, Mn, and Co. The presence of Fe increased the exopolysaccharide precipitation by aluminum. No precipitation, gelation, or increase in turbidity of polysaccharide solutions occurred when K, Na, Ca, Mg, Cu, Cd, Pb, Zn, Hg, or U was added at several pH values. The results suggest that the precipitation is based on the interaction between carboxylate groups from different polysaccharide chains and the partially hydrolyzed aquoions of Fe, Al, Th, and Sn. PMID- 16349467 TI - Analysis of Expression of a Phenazine Biosynthesis Locus of Pseudomonas aureofaciens PGS12 on Seeds with a Mutant Carrying a Phenazine Biosynthesis Locus Ice Nucleation Reporter Gene Fusion. AB - A derivative of Pseudomonas aureofaciens PGS12 expressing a promoterless ice nucleation gene under the control of a phenazine biosynthesis locus was used to study the expression of a phenazine antibiotic locus (Phz) during bacterial seed colonization. Seeds of various plants were inoculated with wild-type PGS12 and a PGS12 ice nucleation-active phz:inaZ marker exchange derivative and planted in soil, and the expression of the reporter gene was monitored at different intervals for 48 h during seed germination. phz gene expression was first detected 12 h after planting, and the expression increased during the next 36-h period. Significant differences in expression of bacterial populations on different seeds were measured at 48 h. The highest expression level was recorded for wheat seeds (one ice nucleus per 4,000 cells), and the lowest expression level was recorded for cotton seeds (one ice nucleus per 12,000,000 cells). These values indicate that a small proportion of bacteria in a seed population expressed phenazine biosynthesis. Reporter gene expression levels and populations on individual seeds in a sample were lognormally distributed. There was greater variability in reporter gene expression than in population size among individual seeds in a sample. Expression on sugar beet and radish seeds was not affected by different inoculum levels or soil matric potentials of -10 and -40 J/kg; only small differences in expression on wheat and sugar beet seeds were detected when the seeds were planted in various soils. It is suggested that the nutrient level in seed exudates is the primary reason for the differences observed among seeds. The lognormal distribution of phenazine expression on seeds and the timing and difference in expression of phenazine biosynthesis on seeds have implications for the potential efficacy of biocontrol microorganisms against plant pathogens. PMID- 16349468 TI - Detection of beta-Glucosidase Activity in Polyacrylamide Gels with Esculin as Substrate. AB - beta-Glucosidase can be located after nondenaturing polyacrylamide gel electrophoresis by incubating the gel with 0.1% esculin and 0.03% ferric chloride. The esculetin released from esculin by beta-glucosidase action reacts with ferric ion to produce a black band, corresponding to the beta-glucosidase, against the transparent background. PMID- 16349469 TI - Transformation of the Fungal Soybean Pathogen Cercospora kikuchii with the Selectable Marker bar. AB - An improved transformation protocol, utilizing selection for resistance to the herbicide bialaphos, has been developed for the plant pathogenic fungus Cercospora kikuchii. Stable, bialaphos-resistant transformants are recovered at frequencies eight times higher than those achieved with the previous system that was based on selection for benomyl resistance. In addition to C. kikuchii, this improved method can also be used to transform other species of Cercospora. PMID- 16349470 TI - Nitrogen Assimilation: Optimization of N-Labelled Azoverdin Production by Azomonas macrocytogenes ATCC 12334. AB - The effects of various nitrogen sources and of the cultivation time on the biosynthesis of N-labelled azoverdin were studied in batch cultures of Azomonas macrocytogenes ATCC 12334. Growth for 48 h in the presence of ammonium sulfate (0.2 g/liter) resulted in 95% labelling of azoverdin with N. A growth-related synthesis of azoverdin, under both nitrogen-fixing and ammonium-assimilating conditions, was observed. PMID- 16349471 TI - Cross-Resistance to Bacillus thuringiensis Toxin CryIF in the Diamondback Moth (Plutella xylostella). AB - Selection with Bacillus thuringiensis subsp. kurstaki, which contains CryIA and CryII toxins, caused a >200-fold cross-resistance to CryIF toxin from B. thuringiensis subsp. aizawai in the diamondback moth, Plutella xylostella. CryIE was not toxic, but CryIB was highly toxic to both selected and unselected larvae. The results show that extremely high levels of cross-resistance can be conferred across classes of CryI toxins of B. thuringiensis. PMID- 16349472 TI - Purification and Characterization of Two Epoxide Hydrolases from Corynebacterium sp. Strain N-1074. AB - Enzymes II(a) and II(b), which catalyze the conversion of epichlorohydrin (ECH) to 3-chloro-1,2-propanediol (MCP), were purified from Corynebacterium sp. strain N-1074, which catalyzes the formation of (R)-MCP from prochiral 1,3-dichloro-2 propanol via ECH. The specific activity of enzyme II(a) for the formation of MCP from ECH was about 6.4-fold higher than that of enzyme II(b). Both enzymes catalyzed the conversion of 1,2-epoxides to the corresponding diol, although they differed in several enzymatic properties. PMID- 16349473 TI - Production and Secretion of the Polysaccharide Biodispersan of Acinetobacter calcoaceticus A2 in Protein Secretion Mutants. AB - Biodispersan is an extracellular anionic polysaccharide produced by Acinetobacter calcoaceticus A2 that changes the surface properties of limestone and acts both as a dispersant and as a grinding aid (E. Rosenberg, C. Rubinovitz, A. Gottlieb, S. Rosenhak, and E. Z. Ron, Appl. Environ. Microbiol. 54:317-322, 1988; E. Rosenberg, C. Rubinovitz, R. Legmann, and E. Z. Ron, Appl. Environ. Microbiol. 54:323-326, 1988; E. Rosenberg, Z. Schwartz, A. Tenenbaum, C. Rubinovitz, R. Legmann, and E. Z. Ron, J. Dispersion Sci. Technol. 10:241-250, 1989). Extracellular fluid also contains a high concentration of secreted proteins that create problems in the purification and application of biodispersan. In order to obtain preparations of biodispersan that contained smaller amounts of protein, we selected mutants of strain A2 that were defective in protein secretion. These mutants produced equal, or even higher, levels of total biodispersan compared with those of the parental strain. Moreover, although there was a significant drop in the concentration of extracellular proteins in the medium, the secretion of biodispersan was unaffected. These results suggest that secretion mutants are potentially useful for the production of extracellular polysaccharides. PMID- 16349474 TI - Reductive Dechlorination of Trichloroethylene and Tetrachloroethylene under Aerobic Conditions in a Sediment Column. AB - [This corrects the article on p. 2202 in vol. 60.]. PMID- 16349475 TI - In Vivo Characterization of Dimethylsulfoniopropionate Lyase in the Fungus Fusarium lateritium. AB - A fungus, Fusarium lateritium, with dimethylsulfoniopropionate (DMSP) lyase activity was isolated from both seawater and a salt marsh due to its ability to grow on DMSP (with the evolution of dimethyl sulfide) as the sole source of carbon. This is the first reported case of DMSP lyase activity in a fungus. Several other common fungal genera tested did not have DMSP lyase activity. DMSP was taken up more rapidly by F. lateritium than it was utilized, leading to its intracellular accumulation. Inhibitor studies with nystatin and cyanide indicated that DMSP uptake was an energy-dependent process. The lyase was inducible by its substrate, DMSP (K(m), 1.2 mM), and by the substrate analogs choline and glycine betaine. During induction, DMSP lyase activity increased with time and then dropped rapidly. This loss of activity could be prevented by spiking the culture with fresh DMSP or choline. The V(max) for DMSP lyase was 34.7 mU . mg of protein. The inhibitory effects of nystatin, and p-chloromercuriphenylsulfonate on DMSP lyase activity suggested that the enzyme is cytosolic. Because plants like Spartina (a marsh grass) and marine algae contain high concentrations of DMSP, we speculate that DMSP-utilizing fungi may be involved in their decay. PMID- 16349477 TI - Microbial consumption of atmospheric isoprene in a temperate forest soil. AB - Isoprene (2-methyl-1,3 butadiene) is a low-molecular-weight hydrocarbon emitted in large quantities to the atmosphere by vegetation and plays a large role in regulating atmospheric chemistry. Until now, the atmosphere has been considered the only significant sink for isoprene. However, in this study we performed both in situ and in vitro experiments with soil from a temperate forest near Ithaca, N.Y., that indicate that the soil provides a sink for atmospheric isoprene and that the consumption of isoprene is carried out by microorganisms. Consumption occurred rapidly in field chambers (672.60 +/- 30.12 to 2,718.36 +/- 86.40 pmol gdw day) (gdw is grams [dry weight] of soil; values are means +/- standard deviations). Subsequent laboratory experiments confirmed that isoprene loss was due to biological processes: consumption was stopped by autoclaving the soil; consumption rates increased with repeated exposure to isoprene; and consumption showed a temperature response consistent with biological activity (with an optimum temperature of 30 degrees C). Isoprene consumption was diminished under low oxygen conditions (120 +/- 7.44 versus 528.36 +/- 7.68 pmol gdw day under ambient O(2) concentrations) and showed a strong relationship with soil moisture. Isoprene-degrading microorganisms were isolated from the site, and abundance was calculated as 5.8 x 10 +/- 3.2 x 10 cells gdw. Our results indicate that soil may provide a significant biological sink for atmospheric isoprene. PMID- 16349476 TI - Purification and Characterization of O-Methyltransferase I Involved in Conversion of Demethylsterigmatocystin to Sterigmatocystin and of Dihydrodemethylsterigmatocystin to Dihydrosterigmatocystin during Aflatoxin Biosynthesis. AB - O-Methyltransferase I, which catalyzes conversions both of demethylsterigmatocystin (DMST) to sterigmatocystin (ST) and of dihydrodemethylsterigmatocystin (DHDMST) to dihydrosterigmatocystin (DHST) during aflatoxin biosynthesis, was purified to apparent homogeneity from the cytosol fraction of the mycelia of Aspergillus parasiticus NIAH-26 through the following chromatography series: phenyl-Sepharose, DEAE-Sepharose, phenyl-Sepharose, Sephacryl S-300, and Matrex gel Green A. The apparent molecular mass was estimated at 150 kDa based on Sephacryl S-300 gel filtration chromatography, and the denaturing molecular mass was 43 kDa based on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The pI of the enzyme was 4.4, and the optimal pH for activity was broad, from 6.5 to 9.0. In competition experiments using the purified enzyme, the formation of ST from DMST was suppressed when DHDMST was added to the reaction mixture and DHST was newly formed. These results indicate that DMST and DHDMST commonly serve as substrates for the enzyme. The K(m) of the enzyme for DMST was 0.94 muM, and that for DHDMST was 2.5 muM. Interestingly, MT I kinetics deviated substantially from standard Michaelis-Menten kinetics, demonstrating substrate inhibition at a higher substrate concentration. PMID- 16349478 TI - Molecular basis of a bacterial consortium: interspecies catabolism of atrazine. AB - Pseudomonas sp. strain ADP contains the genes, atzA, -B, and -C, that encode three enzymes which metabolize atrazine to cyanuric acid. Atrazine-catabolizing pure cultures isolated from around the world contain genes homologous to atzA, B, and -C. The present study was conducted to determine whether the same genes are present in an atrazine-catabolizing bacterial consortium and how the genes and metabolism are subdivided among member species. The consortium contained four or more bacterial species, but two members, Clavibacter michiganese ATZ1 and Pseudomonas sp. strain CN1, collectively mineralized atrazine. C. michiganese ATZ1 released chloride from atrazine, produced hydroxyatrazine, and contained a homolog to the atzA gene that encoded atrazine chlorohydrolase. C. michiganese ATZ1 stoichiometrically metabolized hydroxyatrazine to N-ethylammelide and contained genes homologous to atzB and atzC, suggesting that either a functional AtzB or -C catalyzed N-isopropylamine release from hydroxyatrazine. C. michiganese ATZ1 grew on isopropylamine as its sole carbon and nitrogen source, explaining the ability of the consortium to use atrazine as the sole carbon and nitrogen source. A second consortium member, Pseudomonas sp. strain CN1, metabolized the N-ethylammelide produced by C. michiganese ATZ1 to transiently form cyanuric acid, a reaction catalyzed by AtzC. A gene homologous to the atzC gene of Pseudomonas sp. strain ADP was present, as demonstrated by Southern hybridization and PCR. Pseudomonas sp. strain CN1, but not C. michiganese, metabolized cyanuric acid. The consortium metabolized atrazine faster than did C. michiganese individually. Additionally, the consortium metabolized a much broader set of triazine ring compounds than did previously described pure cultures in which the atzABC genes had been identified. These data begin to elucidate the genetic and metabolic bases of catabolism by multimember consortia. PMID- 16349479 TI - Effect of inoculation techniques and relative humidity on the growth of molds on the surfaces of yellow layer cakes. AB - FOUR INOCULATION TECHNIQUES WERE COMPARED FOR INITIATION OF GROWTH ON CAKE SURFACES: spot, air cabinet, spray (atomizer), and talc addition methods. Molds were isolated from commercial cakes and were identified as Aspergillus sydowii, Aspergillus ochraceus, Penicillium funiculosum, and Eurotium herbariorum. Cake surfaces were inoculated with mold spores and incubated under three equilibrium relative humidity (ERH) levels: 97, 85, and 75%. Random contamination by spores in a ventilated air cabinet was the simplest method of inoculation, but standard deviations in the inoculation rates (20% on a relative scale) were almost twice those observed with the other methods. The spot method was the most reproducible. Cake samples inoculated in the air cabinet had colony counts 10 times lower than those obtained for potato dextrose agar plates at 97% ERH, which was not the case with the spray and talc methods. Growth of molds was much slower in the samples incubated in 75% relative humidity, with all methods. Colony counts were generally similar in systems adjusted at 85 to 97% ERH but were lower for samples incubated at 75% ERH. In comparisons of the shelf life estimates obtained by the various inoculation methods, a correlation coefficient (r) of 0.70 was obtained between the spot method and the other methods of inoculation, while talc, air cabinet, and spray shelf life data were correlated better (r approximately 0.97). The spot method appeared to be the method of choice in consideration of ease of use, precision, and the ability to enable the study of the effects of the environment on mold-free shelf life as well as on the rate of growth of molds on cakes. PMID- 16349480 TI - Carbohydrate and Amino Acid Fermentation in the Free-Living Primitive Protozoon Hexamita sp. AB - Hexamita sp. is an amitochondriate free-living diplomonad which inhabits O(2) limited environments, such as the deep waters and sediments of lakes and marine basins. C nuclear magnetic resonance spectroscopy reveals ethanol, lactate, acetate, and alanine as products of glucose fermentation under microaerobic conditions (23 to 34 muM O(2)). Propionic acid and butyric acid were also detected and are believed to be the result of fermentation of alternative substrates. Production of organic acids was greatest under microaerobic conditions (15 muM O(2)) and decreased under anaerobic (<0.25 muM O(2)) and aerobic (200 to 250 muM O(2)) conditions. Microaerobic incubation resulted in the production of high levels of oxidized end products (70% acetate) compared to that produced under anoxic conditions (20% acetate). In addition, data suggest that Hexamita cells contain the arginine dihydrolase pathway, generating energy from the catabolism of arginine to citrulline, ornithine, NH(4), and CO(2). The rate of arginine catabolism was higher under anoxic conditions than under microaerobic conditions. Hexamita cells were able to grow in the absence of a carbohydrate source, albeit with a lower growth rate and yield. PMID- 16349481 TI - Effect of trichloroethylene on the competitive behavior of toluene-degrading bacteria. AB - The influence of trichloroethylene (TCE) on a mixed culture of four different toluene-degrading bacterial strains (Pseudomonas putida mt-2, P. putida F1, P. putida GJ31, and Burkholderia cepacia G4) was studied with a fed-batch culture. The strains were competing for toluene, which was added at a very low rate (31 nmol mg of cells [dry weight] h). All four strains were maintained in the mixed culture at comparable numbers when TCE was absent. After the start of the addition of TCE, the viabilities of B. cepacia G4 and P. putida F1 and GJ31 decreased 50- to 1,000-fold in 1 month. These bacteria can degrade TCE, although at considerably different rates. P. putida mt-2, which did not degrade TCE, became the dominant organism. Kinetic analysis showed that the presence of TCE caused up to a ninefold reduction in the affinity for toluene of the three disappearing strains, indicating that inhibition of toluene degradation by TCE occurred. While P. putida mt-2 took over the culture, mutants of this strain which could no longer grow on p-xylene arose. Most of them had less or no meta cleavage activity and were able to grow on toluene with a higher growth rate. The results indicate that cometabolic degradation of TCE has a negative effect on the maintenance and competitive behavior of toluene-utilizing organisms that transform TCE. PMID- 16349482 TI - PCR Detection of Cyclic Lipodepsinonapeptide-Producing Pseudomonas syringae pv. syringae and Similarity of Strains. AB - Many strains of Pseudomonas syringae pv. syringae produce one of four classes of small cyclic lipodepsinonapeptides: syringomycins, syringostatins, syringotoxins, or pseudomycins. These metabolites are phytotoxic and growth inhibitory against a broad spectrum of fungi. Their production is dependent upon the expression of conserved biosynthesis and export genes syrB and syrD, respectively. PCR and oligonucleotide primers specific for a 752-bp fragment of syrB were used to identify cyclic lipodepsinonapeptide-producing strains of P. syringae pv. syringae. In contrast, PCR amplification with primers based on syrD did not always correlate with possession of the syrD gene, as indicated by Southern blot analysis, or with cyclic lipodepsinonapeptide production. Sequence comparisons of 400 nucleotides from the syrB PCR-amplified fragments showed 94% plot similarity among 27 strains. In a sequence phenogram, syringostatin and syringotoxin producers were grouped apart from syringomycin-producing strain B301D, with sequences that differed by eight and nine conserved base substitutions, respectively. PCR amplification of the 752-bp syrB fragment offers rapid and accurate detection of cyclic lipodepsinonapeptide-producing strains, and its sequence provides some predictive capabilities for identifying syringotoxin and syringostatin producers. PMID- 16349483 TI - Effect of metal-rich sludge amendments on the soil microbial community. AB - The effects of heavy-metal-containing sewage sludge on the soil microbial community were studied in two agricultural soils of different textures, which had been contaminated separately with three predominantly single metals (Cu, Zn, and Ni) at two different levels more than 20 years ago. We compared three community based microbiological measurements, namely, phospholipid fatty acid (PLFA) analysis to reveal changes in species composition, the Biolog system to indicate metabolic fingerprints of microbial communities, and the thymidine incorporation technique to measure bacterial community tolerance. In the Luddington soil, bacterial community tolerance increased in all metal treatments compared to an unpolluted-sludge-treated control soil. Community tolerance to specific metals increased the most when the same metal was added to the soil; for example, tolerance to Cu increased most in Cu-polluted treatments. A dose-response effect was also evident. There were also indications of cotolerance to metals whose concentration had not been elevated by the sludge treatment. The PLFA pattern changed in all metal treatments, but the interpretation was complicated by the soil moisture content, which also affected the results. The Biolog measurements indicated similar effects of metals and moisture to the PLFA measurements, but due to high variation between replicates, no significant differences compared to the uncontaminated control were found. In the Lee Valley soil, significant increases in community tolerance were found for the high levels of Cu and Zn, while the PLFA pattern was significantly altered for the soils with high levels of Cu, Ni, and Zn. No effects on the Biolog measurements were found in this soil. PMID- 16349484 TI - Initial reductive reactions in aerobic microbial metabolism of 2,4,6 trinitrotoluene. AB - Because of its high electron deficiency, initial microbial transformations of 2,4,6-trinitrotoluene (TNT) are characterized by reductive rather than oxidation reactions. The reduction of the nitro groups seems to be the dominating mechanism, whereas hydrogenation of the aromatic ring, as described for picric acid, appears to be of minor importance. Thus, two bacterial strains enriched with TNT as a sole source of nitrogen under aerobic conditions, a gram-negative strain called TNT-8 and a gram-positive strain called TNT-32, carried out nitro group reduction. In contrast, both a picric acid-utilizing Rhodococcus erythropolis strain, HL PM-1, and a 4-nitrotoluene-utilizing Mycobacterium sp. strain, HL 4-NT-1, possessed reductive enzyme systems, which catalyze ring hydrogenation, i.e., the addition of a hydride ion to the aromatic ring of TNT. The hydride-Meisenheimer complex thus formed (H-TNT) was further converted to a yellow metabolite, which by electrospray mass and nuclear magnetic resonance spectral analyses was established as the protonated dihydride-Meisenheimer complex of TNT (2H-TNT). Formation of hydride complexes could not be identified with the TNT-enriched strains TNT-8 and TNT-32, or with Pseudomonas sp. clone A (2NT), for which such a mechanism has been proposed. Correspondingly, reductive denitration of TNT did not occur. PMID- 16349485 TI - Effects of Ammonium and Non-Ammonium Salt Additions on Methane Oxidation by Methylosinus trichosporium OB3b and Maine Forest Soils. AB - Additions of ammonium and non-ammonium salts inhibit atmospheric methane consumption by soil at salt concentrations that do not significantly affect the soil water potential. The response of soils to non-ammonium salts has previously raised questions about the mechanism of ammonium inhibition. Results presented here show that inhibition of methane consumption by non-ammonium salts can be explained in part by ion-exchange reactions: cations desorb ammonium, with the level of desorption varying as a function of both the cation and anion added; differential desorption results in differential inhibition levels. Differences in the extent of inhibition among ammonium salts can also be explained in part by the effects of anions on ammonium exchange. In contrast, only minimal effects of cations and anions are observed in liquid cultures of Methylosinus trichosporium OB3b. The comparable level of inhibition by equinormal concentrations of NH(4)Cl and (NH(4))(2)SO(4) and the insensitivity of salt inhibition to increasing methane concentrations (from 10 to 100 ppm) are of particular interest, since both of these patterns are in contrast to results for soils. The greater inhibition of methane consumption for NH(4)Cl than (NH(4))(2)SO(4) in soils can be attributed to increased ammonium adsorption by sulfate; increasing inhibition by non-ammonium salts with increasing methane concentrations can be attributed to desorbed ammonium and a physiological mechanism proposed previously for pure cultures. PMID- 16349486 TI - Nitrospira-like bacteria associated with nitrite oxidation in freshwater aquaria. AB - Oxidation of nitrite to nitrate in aquaria is typically attributed to bacteria belonging to the genus Nitrobacter which are members of the alpha subdivision of the class Proteobacteria. In order to identify bacteria responsible for nitrite oxidation in aquaria, clone libraries of rRNA genes were developed from biofilms of several freshwater aquaria. Analysis of the rDNA libraries, along with results from denaturing gradient gel electrophoresis (DGGE) on frequently sampled biofilms, indicated the presence of putative nitrite-oxidizing bacteria closely related to other members of the genus Nitrospira. Nucleic acid hybridization experiments with rRNA from biofilms of freshwater aquaria demonstrated that Nitrospira-like rRNA comprised nearly 5% of the rRNA extracted from the biofilms during the establishment of nitrification. Nitrite-oxidizing bacteria belonging to the alpha subdivision of the class Proteobacteria (e.g., Nitrobacter spp.) were not detected in these samples. Aquaria which received a commercial preparation containing Nitrobacter species did not show evidence of Nitrobacter growth and development but did develop substantial populations of Nitrospira-like species. Time series analysis of rDNA phylotypes on aquaria biofilms by DGGE, combined with nitrite and nitrate analysis, showed a correspondence between the appearance of Nitrospira-like bacterial ribosomal DNA and the initiation of nitrite oxidation. In total, the data suggest that Nitrobacter winogradskyi and close relatives were not the dominant nitrite-oxidizing bacteria in freshwater aquaria. Instead, nitrite oxidation in freshwater aquaria appeared to be mediated by bacteria closely related to Nitrospira moscoviensis and Nitrospira marina. PMID- 16349487 TI - Fingerprinting of cyanobacteria based on PCR with primers derived from short and long tandemly repeated repetitive sequences. AB - The presence of repeated DNA (short tandemly repeated repetitive [STRR] and long tandemly repeated repetitive [LTRR]) sequences in the genome of cyanobacteria was used to generate a fingerprint method for symbiotic and free-living isolates. Primers corresponding to the STRR and LTRR sequences were used in the PCR, resulting in a method which generate specific fingerprints for individual isolates. The method was useful both with purified DNA and with intact cyanobacterial filaments or cells as templates for the PCR. Twenty-three Nostoc isolates from a total of 35 were symbiotic isolates from the angiosperm Gunnera species, including isolates from the same Gunnera species as well as from different species. The results show a genetic similarity among isolates from different Gunnera species as well as a genetic heterogeneity among isolates from the same Gunnera species. Isolates which have been postulated to be closely related or identical revealed similar results by the PCR method, indicating that the technique is useful for clustering of even closely related strains. The method was applied to nonheterocystus cyanobacteria from which a fingerprint pattern was obtained. PMID- 16349488 TI - Choline Derivatives Involved in Osmotolerance of Penicillium fellutanum. AB - Penicillium fellutanum is osmotolerant and xerotolerant when cultured in a low phosphate medium containing 3 M NaCl. Glycerol and erythritol accumulated in cultures with NaCl concentrations up to 2 M; glycerol was the only detectable polyol in cultures containing 3 M NaCl. In cultures with 3 M NaCl, the intracellular levels of glycine betaine and choline-O-sulfate were 22- and 2.6 fold greater (70 and 46 mM), respectively, than those of cultures without added NaCl. The levels of glycine betaine and glycerol decreased in mycelia transferred from a medium containing 3 M NaCl into a fresh medium without added NaCl. NaCl at 3 M inhibited mycelial mass accumulation; this inhibition was partially corrected by supplementation of cultures with glycine betaine (2 mM) or choline-O-sulfate (10 mM). The presence of exogenous choline chloride (2 mM) in plate cultures protected the cells from stress from 3 M NaCl. The data suggest that glycine betaine and choline-O-sulfate are secondary osmoprotectants which are effective at the point that the cell is incapable of synthesizing more glycerol. PMID- 16349489 TI - Effects of UV-A (320 to 399 Nanometers) on Grazing Pressure of a Marine Heterotrophic Nanoflagellate on Strains of the Unicellular Cyanobacteria Synechococcus spp. AB - In the open ocean, where turbidity is very low, UV radiation may be an important factor regulating interactions among planktonic microorganisms. The effect of exposure to UV radiation on grazing by a commonly isolated marine heterotrophic nanoflagellate, Paraphysomonas bandaiensis, on two strains of the cyanobacteria Synechococcus spp. was investigated. Laboratory cultures were exposed to a range of irradiances of artificially produced UV-B (290 to 319 nm) and UV-A (320 to 399 nm) for up to 10 h. At a UV-B irradiance of 0.19 W m, but not 0.12 W m, grazing mortality of Synechococcus spp. and nanoflagellate-specific grazing rates were reduced compared to mortality and grazing rates with UV-A treatment. Within 6 h of exposure, UV-A alone suppressed grazing mortality at irradiances as low as 3.02 W m. The extent to which grazing mortality and nanoflagellate-specific grazing rates were suppressed by UV-A increased with both irradiance and duration of exposure. Over a 6-h exposure period, differences in grazing mortality were largely attributable to differential survival of nanoflagellates. Over a longer period of exposure, there was impairment by UV-A alone of nanoflagellate-specific grazing rates. Rates of primary productivity of Synechococcus spp. were also reduced by UV-A. The extent to which Synechococcus productivity was reduced, compared to the reduction in Synechococcus grazing mortality, depended on the duration of UV-A exposure. These results support the hypothesis that UV-A alone influences the composition and biomass of marine microbial communities by affecting predator-prey interactions and primary production. PMID- 16349490 TI - Isolation of aerobic anoxygenic photosynthetic bacteria from black smoker plume waters of the juan de fuca ridge in the pacific ocean. AB - A strain of the aerobic anoxygenic photosynthetic bacteria was isolated from a deep-ocean hydrothermal vent plume environment. The in vivo absorption spectra of cells indicate the presence of bacteriochlorophyll a incorporated into light harvesting complex I and a reaction center. The general morphological and physiological characteristics of this new isolate are described. PMID- 16349491 TI - Comparison of Energy and Growth Yields for Desulfitobacterium dehalogenans during Utilization of Chlorophenol and Various Traditional Electron Acceptors. AB - Desulfitobacterium dehalogenans grew with formate as the electron donor and 3 chloro-4-hydroxyphenylacetate (3-Cl-4-OHPA) as the electron acceptor, yielding Y(X/formate), Y(X/2e), and Y(X/ATP) ranging from 3.2 to 11.3 g of biomass (dry weight)/mol, thus indicating that energy was conserved through reductive dechlorination. Pyruvate was utilized as the electron donor and acceptor, yielding stoichiometric amounts of acetate and lactate, respectively, and a Y(X/reduced acceptor) of 13.0 g of biomass (dry weight)/mol. The supplementation of pyruvate-containing medium with additional electron acceptors, such as 3-Cl-4 OHPA, nitrate, fumarate, or sulfite, caused pyruvate to be replaced as the electron acceptor and nearly doubled the Y(X/ATP) (Y(X/acetate formed)). A comparison of the yields for 3-Cl-4-OHPA with those for other traditional electron acceptors indicates that the dehalogenation reaction led to the formation of similar amounts of energy equivalents. The various electron acceptors were used concomitantly with 3-Cl-4-OHPA in nonacclimated cultures, but the utilization rates and amounts utilized differed. PMID- 16349492 TI - Stable-carbon-isotope composition of Fatty acids in hydrothermal vent mussels containing methanotrophic and thiotrophic bacterial endosymbionts. AB - Fatty acid biomarker analysis coupled with gas chromatography-isotope ratio mass spectrometry was used to confirm the presence of methanotrophic and thiotrophic bacterial endosymbionts in the tissues of a hydrothermal vent mussel (Bathymodiolus sp.), collected from the Menez Gwen vent field on the mid-Atlantic ridge. Monounsaturated (n-8) fatty acids, which are diagnostic of methanotrophic bacteria, were detected in all three types of tissues examined (gill, posterior adductor, and mantle), although levels were highest in gill tissues where the bacteria were found. Stable-carbon-isotope compositions (delta-C per mille relative to that of Peedee belemnite) of fatty acids for all three tissues ranged from -24.9 to -34.9 per thousand, which encompasses the range predicted for both thiotroph- and methanotroph-based nutrition. The data suggest that these thio- and methanotrophic bacterial endosymbionts are equally important in the nutrition of the vent mussel at this particular vent site. PMID- 16349493 TI - Production of a polyhydroxyalkanoate biopolymer in insect cells with a modified eucaryotic Fatty Acid synthase. AB - [This corrects the article on p. 2540 in vol. 62, PMID: 8779593.]. PMID- 16349494 TI - Purification and Properties of a Polyester Polyurethane-Degrading Enzyme from Comamonas acidovorans TB-35. AB - A polyester polyurethane (PUR)-degrading enzyme, PUR esterase, derived from Comamonas acidovorans TB-35, a bacterium that utilizes polyester PUR as the sole carbon source, was purified until it showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This enzyme was bound to the cell surface and was extracted by addition of 0.2% N,N-bis(3-d gluconamidopropyl)deoxycholamide (deoxy-BIGCHAP). The results of gel filtration and SDS-PAGE showed that the PUR esterase was a monomer with a molecular mass of about 62,000 Da. This enzyme, which is a kind of esterase, degraded solid polyester PUR, with diethylene glycol and adipic acid released as the degradation products. The optimum pH for this enzyme was 6.5, and the optimum temperature was 45 degrees C. PUR degradation by the PUR esterase was strongly inhibited by the addition of 0.04% deoxy-BIGCHAP. On the other hand, deoxy-BIGCHAP did not inhibit the activity when p-nitrophenyl acetate, a water-soluble compound, was used as a substrate. These observations indicated that this enzyme degrades PUR in a two step reaction: hydrophobic adsorption to the PUR surface and hydrolysis of the ester bond of PUR. PMID- 16349495 TI - Manganese Peroxidase-Dependent Oxidation of Glyoxylic and Oxalic Acids Synthesized by Ceriporiopsis subvermispora Produces Extracellular Hydrogen Peroxide. AB - The ligninolytic system of the basidiomycete Ceriporiopsis subvermispora is composed of manganese peroxidase (MnP) and laccase. In this work, the source of extracellular hydrogen peroxide required for MnP activity was investigated. Our attention was focused on the possibility that hydrogen peroxide might be generated by MnP itself through the oxidation of organic acids secreted by the fungus. Both oxalate and glyoxylate were found in the extracellular fluid of C. subvermispora cultures grown in chemically defined media, where MnP is also secreted. The in vivo oxidation of oxalate was measured; CO(2) evolution was monitored after addition of exogenous [C]oxalate to cultures at constant specific activity. In standard cultures, evolution of CO(2) from oxalate was maximal at day 6, although the MnP titers were highest at day 12, the oxalate concentration was maximal (2.5 mM) at day 10, and the glyoxylate concentration was maximal (0.24 mM) at day 5. However, in cultures containing low nitrogen levels, in which the pH is more stable, a better correlation between MnP titers and mineralization of oxalate was observed. Both MnP activity and oxidation of [C]oxalate were negligible in cultures lacking Mn(II). In vitro assays confirmed that Mn(II) dependent oxidation of [C]oxalate by MnP occurs and that this reaction is stimulated by glyoxylate at the concentrations found in cultures. In addition, both organic acids supported phenol red oxidation by MnP without added hydrogen peroxide, and glyoxylate was more reactive than oxalate in this reaction. Based on these results, a model is proposed for the extracellular production of hydrogen peroxide by C. subvermispora. PMID- 16349496 TI - Enzymatic Combustion of Aromatic and Aliphatic Compounds by Manganese Peroxidase from Nematoloma frowardii. AB - The direct involvement of manganese peroxidase (MnP) in the mineralization of natural and xenobiotic compounds was evaluated. A broad spectrum of aromatic substances were partially mineralized by the MnP system of the white rot fungus Nematoloma frowardii. The cell-free MnP system partially converted several aromatic compounds, including [U-C]pentachlorophenol ([U-C]PCP), [U-C]catechol, [U-C]tyrosine, [U-C]tryptophan, [4,5,9,10-C]pyrene, and [ring U-C]2-amino-4,6 dinitrotoluene ([C]2-AmDNT), to CO(2). Mineralization was dependent on the ratio of MnP activity to concentration of reduced glutathione (thiol-mediated oxidation), a finding which was demonstrated by using [C]2-AmDNT as an example. At [C]2-AmDNT concentrations ranging from 2 to 120 muM, the amount of released CO(2) was directly proportional to the concentration of [C]2-AmDNT. The formation of highly polar products was also observed with [C]2-AmDNT and [U-C]PCP; these products were probably low-molecular-weight carboxylic acids. Among the aliphatic compounds tested, glyoxalate was mineralized to the greatest extent. Eighty-six percent of the COOH-glyoxalate and 9% of the CHO-glyoxalate were converted to CO(2), indicating that decarboxylation reactions may be the final step in MnP catalyzed mineralization. The extracellular enzymatic combustion catalyzed by MnP could represent an important pathway for the formation of carbon dioxide from recalcitrant xenobiotic compounds and may also have general significance in the overall biodegradation of resistant natural macromolecules, such as lignins and humic substances. PMID- 16349497 TI - Significance of viral lysis and flagellate grazing as factors controlling bacterioplankton production in a eutrophic lake. AB - The effects of viral lysis and heterotrophic nanoflagellate (HNF) grazing on bacterial mortality were estimated in a eutrophic lake (Lake Plusssee in northern Germany) which was separated by a steep temperature and oxygen gradient into a warm and oxic epilimnion and a cold and anoxic hypolimnion. Two transmission electron microscopy-based methods (whole-cell examination and thin sections) were used to determine the frequency of visibly infected cells, and a model was used to estimate bacterial mortality due to viral lysis. Examination of thin sections also showed that between 20.2 and 29.2% (average, 26.1%) of the bacterial cells were empty (ghosts) and thus could not contribute to viral production. The most important finding was that the mechanism for regulating bacterial production shifted with depth from grazing control in the epilimnion to control due to viral lysis in the hypolimnion. We estimated that in the epilimnion viral lysis accounted on average for 8.4 to 41.8% of the summed mortality (calculated by determining the sum of the mortalities due to lysis and grazing), compared to 51.3 to 91.0% of the summed mortality in the metalimninon and 88.5 to 94.2% of the summed mortality in the hypolimnion. Estimates of summed mortality values indicated that bacterial production was controlled completely or almost completely in the epilimnion (summed mortality, 66.6 to 128.5%) and the hypolimnion (summed mortality, 43.4 to 103.3%), whereas in the metalimnion viral lysis and HNF grazing were not sufficient to control bacterial production (summed mortality, 22.4 to 56.7%). The estimated contribution of organic matter released by viral lysis of cells into the pool of dissolved organic matter (DOM) was low; however, since cell lysis products are very likely labile compared to the bulk DOM, they might stimulate bacterial production. The high mortality of bacterioplankton due to viral lysis in anoxic water indicates that a significant portion of bacterial production in the metalimnion and hypolimnion is cycled in the bacterium-virus-DOM loop. This finding has major implications for the fate and cycling of organic nutrients in lakes. PMID- 16349498 TI - Influence of temperature and pressure on the lethality of ultrasound. AB - A specially designed resistometer was constructed, and the lethal effect on Yersinia enterocolitica of ultrasonic waves (UW) at different static pressures (manosonication [MS]) and of combined heat-UW under pressure treatments (manothermosonication [MTS]) was investigated. During MS treatments at 30 degrees C and 200 kPa, the increase in the amplitude of UW of 20 kHz from 21 to 150 mum exponentially decreased decimal reduction time values (D(MS)) from 4 to 0.37 min. When pressure was increased from 0 to 600 kPa at a constant amplitude (150 mum) and temperature (30 degrees C), D(MS) values decreased from 1.52 to 0.20 min. The magnitude of this decrease in D(MS) declined progressively as pressure was increased. The influence of pressure on D(MS) values was greater with increased amplitude of UW. Pressure alone of as much as 600 kPa did not influence the heat resistance of Y. enterocolitica (D(60) = 0.094; z = 5.65). At temperatures of as much as 58 degrees C, the lethality of UW under pressure was greater than that of heat treatment alone at the same temperature. At higher temperatures, this difference disappeared. Heat and UW under pressure seemed to act independently. The lethality of MTS treatments appeared to result from the added effects of UW under pressure and the lethal effect of heat. The individual contributions of heat and of UW under pressure to the total lethal effect of MTS depended on temperature. The inactivating effect of UW was not due to titanium particles eroded from the sonication horn. The addition to the MS media of cysteamine did not increase the resistance of Y. enterocolitica to MS treatment. MS treatment caused cell disruption. PMID- 16349499 TI - Adaptive changes in membrane lipids of barophilic bacteria in response to changes in growth pressure. AB - The lipid compositions of barophilic bacterial strains which contained docosahexaenoic acid (DHA [22:6n-3]) were examined, and the adaptive changes of these compositions were analyzed in response to growth pressure. In the facultatively barophilic strain 16C1, phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) were major components which had the same fatty acid chains. However, in PE, monounsaturated fatty acids such as hexadecenoic acid were major components, and DHA accounted for only 3.7% of the total fatty acids, while in PG, DHA accounted for 29.6% of the total fatty acids. In response to an increase in growth pressure in strain 16C1, the amounts of saturated fatty acids in PE were reduced, and these decreases were mainly balanced by an increase in unsaturated fatty acids, including DHA. In PG, the decrease in saturated fatty acids was mainly balanced by an increase in DHA. Similar adaptive changes in fatty acid composition were observed in response to growth pressure in obligately barophilic strain 2D2. Furthermore, these adaptive changes in response were also observed in response to low temperature in strain 16C1. These results confirm that the general shift from saturated to unsaturated fatty acids including DHA is one of the adaptive changes in response to increases in pressure and suggest that DHA may play a role in maintaining the proper fluidity of membrane lipids under high pressure. PMID- 16349500 TI - Effects of High Pressure on Inactivation Kinetics and Events Related to Proton Efflux in Lactobacillus plantarum. AB - Knowledge of the mechanism of pressure-induced inactivation of microorganisms could be helpful in defining an effective, relatively mild pressure treatment as a means of decontamination, especially in combination with other physical treatments or antimicrobial agents. We have studied the effect of high pressure on Lactobacillus plantarum grown at pH 5.0 and 7.0. The classical inactivation kinetics were compared with a number of events related to the acid-base physiology of the cell, i.e., activity of F(0)F(1) ATPase, intracellular pH, acid efflux, and intracellular ATP pool. Cells grown at pH 5.0 were more resistant to pressures of 250 MPa than were cells grown at pH 7.0. This difference in resistance may be explained by a higher F(0)F(1) ATPase activity, better ability to maintain a DeltapH, or a higher acid efflux of the cells grown at pH 5.0. After pressure treatment at 250 MPa, the F(0)F(1) ATPase activity was decreased, the ability to maintain a DeltapH was reduced, and the acid efflux was impaired. The ATP pool increased initially after mild pressure treatment and finally decreased after prolonged treatment. The observations on acid efflux and the ATP pool suggest that the glycolysis is affected by high pressure later than is the F(0)F(1) ATPase activity. Although functions related to the membrane-bound ATPase activity were impaired, no morphological changes of the membrane could be observed. PMID- 16349501 TI - Altered specificity of lactococcal proteinase p(i) (lactocepin I) in humectant systems reflecting the water activity and salt content of cheddar cheese. AB - By using various humectant systems, the specificity of hydrolysis of alpha(s1)-, beta-, and kappa-caseins by the cell envelope-associated proteinase (lactocepin; EC 3.4.21.96) with type P(1) specificity (i.e., lactocepin I) from Lactococcus lactis subsp. lactis BN1 was investigated at water activities (a(w)) and salt concentrations reflecting those in cheddar type cheese. In the presence of polyethylene glycol 20000 (PEG 20000)-NaCl (a(w) = 0.95), hydrolysis of beta casein resulted in production of the peptides comprising residues 1 to 6 and 47 to 52, which are characteristic of type P(III) enzyme activity (lactocepin III) in buffer. The fragment comprising residues 1 through 166, inclusive (fragment 1 166), which is typical of lactocepin I activity in buffer systems, was not produced. Similarly, peptide 152-160 from kappa-casein, which is usually produced in aqueous buffers exclusively by lactocepin III, was a major product of lactocepin I. Most of the specificity differences obtained in the presence of PEG 20000-NaCl were also obtained in the presence of PEG 20000 alone (a(w) = 0.99). In addition, alpha(s1)-casein, which normally is resistant to lactocepin I activity, was rapidly hydrolyzed in the presence of PEG 20000 alone. Hydrolysis of casein in the presence of PEG 300-NaCl or glycerol-NaCl (both having an a(w) of 0.95) was generally as expected for lactocepin I activity except that beta casein peptide 47-52 and kappa-casein fragment 1-160 were produced; both of these are normally formed by lactocepin III in buffer. The differences in lactocepin specificity obtained in the humectant systems can be attributed to a combination of a(w) and humectant hydrophobicity, both of which are parameters that are potentially relevant to the cheese-ripening environment. PMID- 16349502 TI - Induction of Mannanase, Xylanase, and Endoglucanase Activities in Sclerotium rolfsii. AB - Induction of mannanase, xylanase, and cellulase (endoglucanase) synthesis in the plant-pathogenic basidiomycete Sclerotium rolfsii was studied by incubating noninduced, resting mycelia with a number of mono-, oligo-, and polysaccharides. The simultaneous formation of these three endoglycanases could be provoked by several polysaccharides structurally resembling the carbohydrate constituents of lignocellulose (e.g., mannan and cellulose), by various disaccharide catabolites of these lignocellulose constituents (e.g., cellobiose, mannobiose, and xylobiose), or by structurally related disaccharides (e.g., lactose, sophorose, and galactosyl-beta-1,4-mannose), as well as by l-sorbose. Synthesis of mannanase, xylanase, and endoglucanase always occurred concomitantly and could not be separated by selecting an appropriate inducer. Various structurally different inducing carbohydrates promoted the excretion of the same multiple isoforms of endoglycanases, as judged from the similar banding patterns obtained in zymogram analyses of enzyme preparations obtained in response to these different inducers and resolved by analytical isoelectric focusing. Whereas enhanced xylanase and endoglucanase formation is strictly dependent on the presence of suitable inducers, increased levels of mannanase are excreted by S. rolfsii even under noninducing, derepressed conditions, as shown in growth experiments with glucose as the substrate. Significant mannanase formation commenced only when glucose was exhausted from the medium. Under these conditions, only very low, presumably constitutive levels of xylanase and endoglucanase were formed. Although the induction of the three endoglycanases is very closely related in S. rolfsii, it was concluded that there is no common, coordinated regulatory mechanism that controls the synthesis of mannanase, xylanase, and endoglucanase. PMID- 16349503 TI - Osmoregulatory Responses of Fungi Inhabiting Standing Litter of the Freshwater Emergent Macrophyte Juncus effusus. AB - Standing litter of emergent macrophytes often forms a major portion of the detrital mass in wetland habitats. Microbial assemblages inhabiting this detritus must adapt physiologically to daily fluctuations in temperature and water availability. We examined the effects of various environmental conditions on the concentrations of osmoregulatory solutes (polyols and trehalose) and the respiratory activities of fungal assemblages inhabiting standing litter of the freshwater emergent macrophyte Juncus effusus. Under field conditions, the concentrations of osmolytes (polyols plus trehalose) in fungal decomposers were negatively correlated with plant litter water potentials (r = -0.75, P < 0.001) and rates of microbial respiration (r = -0.66, P < 0.001). The highest concentration of osmolytes (polyols plus trehalose) occurred in standing litter exposed to desiccating conditions (range from wet to dry, 0.06 to 0.68 mumol . mg of fungal biomass). Similar fluctuations in polyol and trehalose concentrations were observed in standing litter wetted and dried under laboratory conditions and for four predominant fungal decomposers of J. effusus grown individually on sterilized Juncus leaves. These studies suggest that fungal inhabitants associated with standing litter of emergent macrophytes can adjust their intracellular solute concentrations in response to daily fluctuations in water availability. PMID- 16349504 TI - Identification of hydrocarbon-degrading bacteria in soil by reverse sample genome probing. AB - Bacteria with limited genomic cross-hybridization were isolated from soil contaminated with C5+, a mixture of hydrocarbons, and identified by partial 16S rRNA sequencing. Filters containing denatured genomic DNAs were used in a reverse sample genome probe (RSGP) procedure for analysis of the effect of an easily degradable compound (toluene) and a highly recalcitrant compound (dicyclopentadiene [DCPD]) on community composition. Hybridization with labeled total-community DNA isolated from soil exposed to toluene indicated enrichment of several Pseudomonas spp., which were subsequently found to be capable of toluene mineralization. Hybridization with labeled total-community DNA isolated from soil exposed to DCPD indicated enrichment of a Pseudomonas sp. or a Sphingomonas sp. These two bacteria appeared capable of producing oxygenated DCPD derivatives in the soil environment, but mineralization could not be shown. These results demonstrate that bacteria, which metabolize degradable or recalcitrant hydrocarbons, can be identified by the RSGP procedure. PMID- 16349505 TI - Metabolism of Dichloromethane by the Strict Anaerobe Dehalobacterium formicoaceticum. AB - The metabolism of dichloromethane by Dehalobacterium formicoaceticum in cell suspensions and crude cell extracts was investigated. The organism is a strictly anaerobic gram-positive bacterium that utilizes exclusively dichloromethane as a growth substrate and ferments this compound to formate and acetate in a molar ratio of 2:1. When [C]dichloromethane was degraded by cell suspensions, formate, the methyl group of acetate, and minor amounts of methanol were labeled, but there was no nuclear magnetic resonance signal corresponding to the carboxyl group of acetate. This finding and previously established carbon and electron balances suggested that dichloromethane was converted to methylene tetrahydrofolate, of which two-thirds was oxidized to formate while one-third gave rise to acetate by incorporation of CO(2) from the medium in the acetyl coenzyme A synthase reaction. When crude desalted extracts were incubated in the presence of dichloromethane, tetrahydrofolate, ATP, methyl viologen, and molecular hydrogen, dichloromethane and tetrahydrofolate were consumed, with the concomitant formation of stoichiometric amounts of methylene tetrahydrofolate. The in vitro transfer of the methylene group of dichloromethane onto tetrahydrofolate required substoichiometric amounts of ATP. The reaction was inhibited in a light-reversible fashion by 20 muM propyl iodide, thus suggesting involvement of a Co(I) corrinoid in the anoxic dehalogenation of dichloromethane. D. formicoaceticum exhibited normal growth with 0.8 mM sodium in the medium, and crude extracts contained ATPase activity that was partially inhibited by N,N' dicyclohexylcarbodiimide and azide. During growth with dichloromethane, the organism thus may conserve energy not only by substrate-level phosphorylation but also by a chemiosmotic mechanism involving a sodium-independent F(0)F(1)-type ATP synthase. PMID- 16349506 TI - Characterization of the Cricket Hindgut Microbiota with Fluorescently Labeled rRNA-Targeted Oligonucleotide Probes. AB - Most cricket hindgut microorganisms (60 to 80%) were detected with a universal fluorescent rRNA-targeted probe and found to be eubacteria. Group-specific probes showed that the hindguts of five different cricket species harbor similar bacterial groups, although in different proportions, and that different diets shifted the structure of the hindgut microbial community. The Bacteroides Prevotella probe, of the eight eubacterial probes tested, stained the largest percentage of cells in all crickets. PMID- 16349507 TI - Differential Gene Expression in the Laccase Gene Family from Basidiomycete I-62 (CECT 20197). AB - A family of genes encoding laccases has recently been described for the basidiomycete I-62 (CECT 20197). Transcript levels of genes lcc1, lcc2, and lcc3 were analyzed under four different culture conditions to study their expression patterns. Two of the laccase genes were clearly inducible by veratryl alcohol: the lcc1 gene is inducible in early stages of growth, and the lcc2 gene is also inducible but only when the organism reaches the stationary phase. Transcript levels for the third gene, lcc3, were uninduced by veratryl alcohol and repressed by glucose. PMID- 16349508 TI - Inactivation of Cryptosporidium parvum Oocysts by Ammonia. AB - The survival of Cryptosporidium parvum oocysts in soil and water microhabitats may be affected by the environmental production and release of free ammonia. The objective of this study was to determine the effects of increasing free ammonia concentrations and times of exposure on oocyst viability. Wild-type oocysts were obtained from naturally infected calf feces by chemical (continuous-flow) centrifugation and sucrose gradients. Ammonia (NH(3)) from a commercial solution was applied in concentrations ranging from 0.007 to 0.148 M. Exposure times ranged from 10 min to 24 h at a constant temperature of 24 +/- 1 degrees C. Viability of oocysts was determined with a dye permeability assay and an in vitro excystation assay (M. B. Jenkins, L. J. Anguish, D. D. Bowman, M. J. Walker, and W. C. Ghiorse, Appl. Environ. Microbiol. 63:3844-3850, 1997). Even the lowest concentration of ammonia decreased significantly the viability of oocysts after 24 h of exposure. Increasing concentrations of ammonia increased inactivation rates, which ranged from 0.014 to 0.066 h. At the highest concentration of ammonia, a small fraction of viable oocysts still remained. Exposure to pH levels corresponding to those associated with the ammonia concentrations showed minimal effects of alkaline pH alone on oocyst viability. This study shows that environmentally relevant concentrations of free ammonia may significantly increase the inactivation of oocysts in ammonia-containing environments. PMID- 16349509 TI - Mercury methylation and demethylation in anoxic lake sediments and by strictly anaerobic bacteria. AB - After spiking anoxic sediment slurries of three acidic oligotrophic lakes with either HgCl(2) at 1.0 mug/ml or CH(3)HgI at 0.1 mug/ml, both mercury methylation and demethylation rates were measured. High mercury methylation potentials were accompanied by high demethylation potentials in the same sediment. These high potentials correlated positively with the concentrations of organic matter and dissolved sulfate in the sediment and with mercury levels in fish. Adjustment of the acidic sediment pH to neutrality failed to influence either the methylation or the demethylation rate of mercury. The opposing methylation and demethylation processes converged to establish similar Hg-CH(3)Hg equilibria in all three sediments. Because of their metabolic dominance in anoxic sediments, mercury methylation and demethylation in pure cultures of sulfidogenic, methanogenic, and acetogenic bacteria were also measured. Sulfidogens both methylated and demethylated mercury, but the methanogen tested only catalyzed demethylation and the acetogen neither methylated nor demethylated mercury. PMID- 16349510 TI - Oxygen Consumption by Desulfovibrio Strains with and without Polyglucose. AB - The kinetics of oxygen reduction by Desulfovibrio salexigens Mast1 and the role of polyglucose in this activity were examined and compared with those of strains of D. desulfuricans and D. gigas. Oxidation rates were highest at air saturation (up to 40 nmol of O(2) min mg of protein) and declined with decreasing oxygen concentrations. Studies with cell extracts (CE) indicated that NADH oxidase was entirely responsible for the oxygen reduction in strain Mast1. In D. desulfuricans CSN, at least three independent systems appeared to reduce oxygen. Two were active at all oxygen concentrations (NADH oxidase and NADPH oxidase), and one was maximally active at less than 10 muM oxygen. In contrast to D. gigas and D. salexigens strains, the D. desulfuricans strains also contained NADH peroxidase and NADPH peroxidase activities and did not accumulate polyglucose under nonlimiting growth conditions. At air saturation, initial activities of the oxidases and peroxidases of cells harvested at the end of the log phase were on the order of 20 to 140 nmol of O(2) min mg of protein. In all strains, these enzymes were relatively stable but were susceptible to inactivation as soon as substrates were added to the assay mixture. Under those conditions, all oxidation activity disappeared after ca. 1 h of incubation. The same finding was observed with whole cells of D. desulfuricans CSN and D. desulfuricans ATCC 27774, but inactivation was less pronounced with cells of D. salexigens Mast1. It appeared that the presence of polyglucose in the whole cells retarded the process of inactivation of NADH oxidase, but this property was lost in crude CE. In spite of the effect of polyglucose on the oxidative potential, oxygen-dependent growth of D. salexigens Mast1 could be demonstrated neither in batch nor in continuous culture. PMID- 16349511 TI - Uptake of choline and its conversion to glycine betaine by bacteria in estuarine waters. AB - The uptake and degradation of nanomolar levels of [methyl-C]choline in estuarine water samples and in seawater filtrate cultures composed mainly of natural free living bacteria was studied. Uptake of [C]choline exhibited Michaelis-Menten kinetics, with K(t) + S(n) values of 1.7 to 2.9 nM in filtrate cultures and 1.7 to 4.1 nM in estuarine-water samples. V(max) values ranged from 0.5 to 3.3 nM . h. The uptake system for choline in natural microbial assemblages therefore displays very high affinity and appears able to scavenge this compound at the concentrations expected in seawater. Uptake of choline was inhibited by some natural structural analogs and p-chloromercuribenzoate, indicating that the transporter may be multifunctional and may involve a thiol binding site. When 11 nM [C]choline was added to water samples, a significant fraction (>50%) of the methyl carbon was respired to CO(2) in incubations lasting 10 to 53 h. Cells taking up [C]choline produced [C]glycine betaine ([C]GBT), and up to 80% of the radioactivity retained by cells was in the form of GBT, a well-known osmolyte. Alteration of the salinity in filtrate cultures affected the relative proportion of [C]choline degraded or converted to [C]GBT, without substantially affecting the total metabolism of choline. Increasing the salinity from 14 to 25 or 35 ppt caused more [C]GBT to be produced from choline but less CO(2) to be produced than in the controls. Lowering the salinity to 7 ppt decreased [C]GBT production and increased CO(2) production slightly. Intracellular accumulations of [C]GBT in the salt-stressed cultures were osmotically significant (34 mM). Choline may be used as an energy substrate by estuarine bacteria and may also serve as a precursor of the osmoprotectant GBT, particularly as bacteria are mixed into higher-salinity waters. PMID- 16349512 TI - Microbial reductive dechlorination of aroclor 1260 in anaerobic slurries of estuarine sediments. AB - Reductive dechlorination of Aroclor 1260 was investigated in anaerobic slurries of estuarine sediments from Baltimore Harbor (Baltimore, Md.). The sediment slurries were amended with 800 ppm Aroclor 1260 with and without the addition of 350 muM 2,3,4,5-tetrachlorobiphenyl (2,3,4,5-CB) or 2,3,5,6-tetrachlorobiphenyl (2,3,5,6-CB) and incubated in triplicate at 30 degrees C under methanogenic conditions in an artificial estuarine medium. After 6 months, extensive meta dechlorination and moderate ortho dechlorination of Aroclor 1260 occurred in all incubated cultures except for sterilized controls. Overall, total chlorines per biphenyl decreased by up to 34%. meta chlorines per biphenyl decreased by 65, 55, and 45% and ortho chlorines declined by 18, 12, and 9%, respectively, when 2,3,4,5-CB, 2,3,5,6-CB, or no additional congener was supplied. This is the first confirmed report of microbial ortho dechlorination of a commercial polychlorinated biphenyl mixture. In addition, compared with incubated cultures supplied with Aroclor 1260 alone, the dechlorination of Aroclor 1260 plus 2,3,4,5 CB or 2,3,5,6-CB occurred with shorter lag times (31 to 60 days versus 90 days) and was more extensive, indicating that the addition of a single congener stimulated the dechlorination of Aroclor 1260. PMID- 16349513 TI - Ultradian Growth in Prochlorococcus spp. AB - Species of the widespread marine prokaryote Prochlorococcus exhibited ultradian growth (faster than 1 division per day) both in situ and in culture, even though cell division is strictly phased to the light-dark cycle. Under optimal conditions a second DNA replication and cell division closely followed, but did not overlap with, the first division. The timing of cell cycle events was not affected by light intensity or duration, suggesting control by a light-triggered timer or circadian clock rather than by completion of a light-dependent assimilation phase. This mode of ultradian growth has not been observed previously and poses new questions about the regulation of cellular rhythms in prokaryotes. In addition, it implies that conclusions regarding the lack of nutrient limitation of Prochlorococcus in the open ocean, which were based on the appearance that cells were growing at their maximal rate, need to be reconsidered. PMID- 16349514 TI - Methanol promotes atmospheric methane oxidation by methanotrophic cultures and soils. AB - Two methanotrophic bacteria, Methylobacter albus BG8 and Methylosinus trichosporium OB3b, oxidized atmospheric methane during batch growth on methanol. Methane consumption was rapidly and substantially diminished (95% over 9 days) when washed cell suspensions were incubated without methanol in the presence of atmospheric methane (1.7 ppm). Methanotrophic activity was stimulated after methanol (10 mM) but not methane (1,000 ppm) addition. M. albus BG8 grown in continuous culture for 80 days with methanol retained the ability to oxidize atmospheric methane and oxidized methane in a chemostat air supply. Methane oxidation during growth on methanol was not affected by methane deprivation. Differences in the kinetics of methane uptake (apparent K(m) and V(max)) were observed between batch- and chemostat-grown cultures. The V(max) and apparent K(m) values (means +/- standard errors) for methanol-limited chemostat cultures were 133 +/- 46 nmol of methane 10 cells h and 916 +/- 235 ppm of methane (1.2 muM), respectively. These values were significantly lower than those determined with batch-grown cultures (V(max) of 648 +/- 195 nmol of methane 10 cells h and apparent K(m) of 5,025 +/- 1,234 ppm of methane [6.3 muM]). Methane consumption by soils was stimulated by the addition of methanol. These results suggest that methanol or other nonmethane substrates may promote atmospheric methane oxidation in situ. PMID- 16349515 TI - Characterization of Root-Associated Methanotrophs from Three Freshwater Macrophytes: Pontederia cordata, Sparganium eurycarpum, and Sagittaria latifolia. AB - Root-associated methanotrophic bacteria were enriched from three common aquatic macrophytes: Pontederia cordata, Sparganium eurycarpum, and Sagittaria latifolia. At least seven distinct taxa belonging to groups I and II were identified and presumptively assigned to the genera Methylosinus, Methylocystis, Methylomonas, and Methylococcus. Four of these strains appeared to be novel on the basis of partial 16S ribosomal DNA sequence analysis. The root-methanotroph association did not appear to be highly specific, since multiple methanotrophs were isolated from each of the three plant species. Group II methanotrophs were isolated most frequently; though less common, group I isolates accounted for three of the seven distinct methanotrophs. Apparent K(m) values for methane uptake by representative cultures ranged from 3 to >17 muM; for five of the eight cultures examined, apparent K(m) values agreed well with apparent K(m) estimates for plant roots, suggesting that these strains may be representative of those active in situ. PMID- 16349516 TI - Methane and Trichloroethylene Degradation by Methylosinus trichosporium OB3b Expressing Particulate Methane Monooxygenase. AB - Whole-cell assays of methane and trichloroethylene (TCE) consumption have been performed on Methylosinus trichosporium OB3b expressing particulate methane monooxygenase (pMMO). From these assays it is apparent that varying the growth concentration of copper causes a change in the kinetics of methane and TCE degradation. For M. trichosporium OB3b, increasing the copper growth concentration from 2.5 to 20 muM caused the maximal degradation rate of methane (V(max)) to decrease from 300 to 82 nmol of methane/min/mg of protein. The methane concentration at half the maximal degradation rate (K(s)) also decreased from 62 to 8.3 muM. The pseudo-first-order rate constant for methane, V(max)/K(s), doubled from 4.9 x 10 to 9.9 x 10 liters/min/mg of protein, however, as the growth concentration of copper increased from 2.5 to 20 muM. TCE degradation by M. trichosporium OB3b was also examined with varying copper and formate concentrations. M. trichosporium OB3b grown with 2.5 muM copper was unable to degrade TCE in both the absence and presence of an exogenous source of reducing equivalents in the form of formate. Cells grown with 20 muM copper, however, were able to degrade TCE regardless of whether formate was provided. Without formate the V(max) for TCE was 2.5 nmol/min/mg of protein, while providing formate increased the V(max) to 4.1 nmol/min/mg of protein. The affinity for TCE also increased with increasing copper, as seen by a change in K(s) from 36 to 7.9 muM. V(max)/K(s) for TCE degradation by pMMO also increased from 6.9 x 10 to 5.2 x 10 liters/min/mg of protein with the addition of formate. From these whole-cell studies it is apparent that the amount of copper available is critical in determining the oxidation of substrates in methanotrophs that are expressing only pMMO. PMID- 16349517 TI - Surface Charge Properties of and Cu(II) Adsorption by Spores of the Marine Bacillus sp. Strain SG-1. AB - Spores of marine Bacillus sp. strain SG-1 are capable of oxidizing Mn(II) and Co(II), which results in the precipitation of Mn(III, IV) and Co(III) oxides and hydroxides on the spore surface. The spores also bind other heavy metals; however, little is known about the mechanism and capacity of this metal binding. In this study the characteristics of the spore surface and Cu(II) adsorption to this surface were investigated. The specific surface area of wet SG-1 spores was 74.7 m per g of dry weight as measured by the methylene blue adsorption method. This surface area is 11-fold greater than the surface area of dried spores, as determined with an N(2) adsorption surface area analyzer or as calculated from the spore dimensions, suggesting that the spore surface is porous. The surface exchange capacity as measured by the proton exchange method was found to be 30.6 mumol m, which is equal to a surface site density of 18.3 sites nm. The SG-1 spore surface charge characteristics were obtained from acid-base titration data. The surface charge density varied with pH, and the zero point of charge was pH 4.5. The titration curves suggest that the spore surface is dominated by negatively charged sites that are largely carboxylate groups but also phosphate groups. Copper adsorption by SG-1 spores was rapid and complete within minutes. The spores exhibited a high affinity for Cu(II). The amounts of copper adsorbed increased from negligible at pH 3 to maximum levels at pH >6. Their great surface area, site density, and affinity give SG-1 spores a high capability for binding metals on their surfaces, as demonstrated by our experiments with Cu(II). PMID- 16349518 TI - Potential of unicellular cyanobacteria from saline environments as exopolysaccharide producers. AB - Fifteen Cyanothece strains isolated from saline environments have been characterized with regard to exopolysaccharide (EPS) production. The polymers contained six to eight monosaccharides, with one or two acidic sugars. In some EPS samples, the additional presence of acetyl, pyruvyl, and/or sulfate groups was also detected. PMID- 16349519 TI - Methanol improves methane uptake in starved methanotrophic microorganisms. AB - Methanotrophs in enrichment cultures grew and sustained atmospheric methane oxidation when supplied with methanol. If they were not supplied with methanol or formate, their atmospheric methane oxidation came to a halt, but it was restored within hours in response to methanol or formate. Indigenous forest soil methanotrophs were also dependent on a supply of methanol upon reduced methane access but only when exposed to a methane-free atmosphere. Their immediate response to each methanol addition, however, was to shut down the oxidation of atmospheric methane and to reactivate atmospheric methane oxidation as the methanol was depleted. PMID- 16349520 TI - Geographical Differentiation of the Population of Xanthomonas axonopodis pv. manihotis in Colombia. AB - [This corrects the article on p. 4430 in vol. 63.]. PMID- 16349521 TI - Studies of the Catabolic Pathway of Degradation of Nitrobenzene by Pseudomonas pseudoalcaligenes JS45: Removal of the Amino Group from 2-Aminomuconic Semialdehyde. AB - [This corrects the article on p. 4841 in vol. 63.]. PMID- 16349522 TI - Degradation of the Fluoroquinolone Enrofloxacin by the Brown Rot Fungus Gloeophyllum striatum: Identification of Metabolites. AB - [This corrects the article on p. 4276 in vol. 63.]. PMID- 16349523 TI - Seasonality in antarctic airborne fungal spores. AB - [This corrects the article on p. 2243 in vol. 63.]. PMID- 16349524 TI - The Vibrio fischeri-Euprymna scolopes Light Organ Association: Current Ecological Paradigms. PMID- 16349525 TI - Light-Mediated Nitrite Accumulation during Denitrification by Pseudomonas sp. Strain JR12. AB - The effect of light on the denitrifying characteristics of a nonphotosynthetic denitrifier, Pseudomonas sp. strain JR12, was examined. Already at low light intensities, nitrite accumulated as a result of light inhibition of nitrite but not of nitrate reduction rates. Exposure of this bacterium to light caused a photooxidation of cytochrome c, an intermediate electron carrier in its respiratory pathway. Photoinhibition of nitrite reduction was reversible, as nitrite reduction rates returned to preillumination levels when light-exposed cells were returned to dark conditions. Antimycin A reversed the inhibitory effect of light on nitrite reduction by preventing a reversed electron flow. Aerobic respiration by this bacterium was not affected by light. PMID- 16349526 TI - 2-chloro-1,4-dimethoxybenzene as a novel catalytic cofactor for oxidation of anisyl alcohol by lignin peroxidase. AB - 2-Chloro-1,4-dimethoxybenzene (2Cl-14DMB) is a natural compound produced de novo by several white rot fungi. This chloroaromatic metabolite was identified as a cofactor superior to veratryl alcohol (VA) in the oxidation of anisyl alcohol (AA) by lignin peroxidase (LiP). Our results reveal that good LiP substrates, such as VA and tryptophan, are comparatively poor cofactors in the oxidation of AA. Furthermore, we show that a good cofactor does not necessarily serve a role in protecting LiP against H(2)O(2) inactivation. 2Cl-14DMB was not a direct mediator of AA oxidation, since increasing AA concentrations did not inhibit the oxidation of 2Cl-14DMB at all. However, the high molar ratio of anisaldehyde formed to 2Cl-14DMB consumed, up to 13:1, indicates that a mechanism which recycles the cofactor is present. PMID- 16349527 TI - Use of an oxygen-insensitive microscale biosensor for methane to measure methane concentration profiles in a rice paddy. AB - An oxygen-insensitive microscale biosensor for methane was constructed by furnishing a previously described biosensor with an oxygen guard. The guard consisted of a glass capillary containing heterotrophic bacteria, which consumed oxygen diffusing through the tip membrane, thus preventing it from diffusing into the methane-sensing unit. Oxygen microprofiles were measured through the oxygen guard capillary, demonstrating the principle and limitations of the method. When the tip of the guard capillary was exposed to 100% oxygen at 21 degrees C, heterotrophic oxygen consumption prevented oxygen from diffusing further than 170 mum into the capillary, whereas atmospheric levels of oxygen were consumed within 50 mum. The capacity of the oxygen guard for scavenging oxygen decreased with decreasing temperature, and atmospheric levels of oxygen caused oxygen penetration to 200 mum at 5 degrees C. The sensors could be manufactured with tip diameters as small as 25 mum, and response times were about 1 min at room temperature. Pore water profiles of methane concentrations in a rice paddy soil were measured, and a strong correlation between the depths of oxygen penetration and methane appearance was observed as a function of the light regimen; this finding confirmed the role of microbenthic photosynthesis in limiting methane emissions from surfaces of waterlogged sediments and soils. PMID- 16349528 TI - Purification and Characterization of Exo-beta-d-Glucosaminidase from a Cellulolytic Fungus, Trichoderma reesei PC-3-7. AB - Chitosan-degrading activities induced by glucosamine (GlcN) or N acetylglucosamine (GlcNAc) were found in a culture filtrate of Trichoderma reesei PC-3-7. One of the chitosan-degrading enzymes was purified to homogeneity by precipitation with ammonium sulfate followed by anion-exchange and hydrophobic interaction chromatographies. The enzyme was monomeric, and its molecular mass was 93 kDa. The optimum pH and temperature of the enzyme were 4.0 and 50 degrees C, respectively. The activity was stable in the pH range 6.0 to 9.0 and at a temperature below 50 degrees C. Reaction product analysis from the viscosimetric assay and thin-layer chromatography and H nuclear magnetic resonance spectroscopy clearly indicated that the enzyme was an exo-type chitosanase, exo-beta-d glucosaminidase, that releases GlcN from the nonreducing end of the chitosan chain. H nuclear magnetic resonance spectroscopy also showed that the exo-beta-d glucosaminidase produced a beta-form of GlcN, demonstrating that the enzyme is a retaining glycanase. Time-dependent liberation of the reducing sugar from partially acetylated chitosan with exo-beta-d-glucosaminidase and the partially purified exo-beta-d-N-acetylglucosaminidase from T. reesei PC-3-7 suggested that the exo-beta-d-glucosaminidase cleaves the glycosidic link of either GlcN-beta(1- >4)-GlcN or GlcN-beta(1-->4)-GlcNAc. PMID- 16349529 TI - Lipopolysaccharide profiles from nodules as markers of bradyrhizobium strains nodulating wild legumes. AB - To develop the use of electrophoretic lipopolysaccharide profiles for Bradyrhizobium strain identification, we studied the feasibility of using electrophoresis of whole legume nodule homogenates to obtain distinctive lipopolysaccharide profiles. The electrophoretic patterns were the same whether we used nodule extracts, bacteroids, or cultured bacteria as samples, and there was no evidence of changes in the ladder-like pattern during the nodulation process. To assess the reliability of using lipopolysaccharide profiling performed with individual nodules for studying the diversity and microdistribution of the rhizobia nodulating wild shrub legumes, we used a population of Adenocarpus foliolosus seedlings. We obtained 75 different profiles from the 147 nodules studied. There was no dominant profile in the sample, and a plant with different nodules generally produced different profiles. Electrophoresis of legume root nodules proved to be a fast and discriminating technique for determining the diversity of a bradyrhizobial population, although it did not allow the genetic relationships among the nodulating strains to be studied. PMID- 16349530 TI - Complete reductive dehalogenation of brominated biphenyls by anaerobic microorganisms in sediment. AB - We sought to determine whether microorganisms from the polychlorinated biphenyl (PCB)-contaminated sediment in Woods Pond (Lenox, Mass.) could dehalogenate brominated biphenyls. The PCB dechlorination specificities for the microorganisms in this sediment have been well characterized. This allowed us to compare the dehalogenation specificities for brominated biphenyls and chlorinated biphenyls within a single sediment. Anaerobic sediment microcosms were incubated separately at 25 degrees C with 16 different mono- to tetrabrominated biphenyls (350 muM) and disodium malate (10 mM). Samples were extracted and analyzed by gas chromatography with an electron capture detector and a mass spectrometer detector at various times for up to 54 weeks. All of the tested brominated biphenyls were dehalogenated. For most congeners, including 2,6-dibromobiphenyl (26-BB) and 24 25-BB, the dehalogenation began within 1 to 2 weeks. However, for 246-BB and 2-2 BB, debromination was first observed at 7 and 14 weeks, respectively. Most intermediate products did not persist, but when 2-2-BB was produced as a dehalogenation product, it persisted for at least 15 weeks before it was dehalogenated to 2-BB and then to biphenyl. The dehalogenation specificities for brominated and chlorinated biphenyls were similar: meta and para substituents were generally removed first, and ortho substituents were more recalcitrant. However, the brominated biphenyls were better dehalogenation substrates than the chlorinated biphenyls. All of the tested bromobiphenyls, including those with ortho and unflanked meta and para substituents, were ultimately dehalogenated to biphenyl, whereas their chlorinated counterparts either were not dehalogenation substrates or were only partially dehalogenated. Our data suggest that PCB dechlorinating microorganisms may be able to dehalogenate brominated biphenyls and may exhibit a relaxed specificity for these substrates. PMID- 16349531 TI - Distribution of a Population of Rhizobium leguminosarum bv. trifolii among Different Size Classes of Soil Aggregates. AB - A combination of the plant infection-soil dilution technique (most-probable number [MPN] technique) and immunofluorescence direct count (IFDC) microscopy was used to examine the effects of three winter cover crop treatments on the distribution of a soil population of Rhizobium leguminosarum bv. trifolii across different size classes of soil aggregates (<0.25, 0.25 to 0.5, 0.5 to 1.0, 1.0 to 2.0, and 2.0 to 5.0 mm). The aggregates were prepared from a Willamette silt loam soil immediately after harvest of broccoli (September 1995) and before planting and after harvest of sweet corn (June and September 1996, respectively). The summer crops were grown in soil that had been either fallowed or planted with a cover crop of red clover (legume) or triticale (cereal) from September to April. The Rhizobium soil population was heterogeneously distributed across the different size classes of soil aggregates, and the distribution was influenced by cover crop treatment and sampling time. On both September samplings, the smallest size class of aggregates (<0.25 mm) recovered from the red clover plots carried between 30 and 70% of the total nodulating R. leguminosarum population, as estimated by the MPN procedure, while the same aggregate size class from the June sampling carried only approximately 6% of the population. In June, IDFC microscopy revealed that the 1.0- to 2.0-mm size class of aggregates from the red clover treatment carried a significantly greater population density of the successful nodule-occupying serotype, AR18, than did the aggregate size classes of <0.5 mm, and 2 to 5 mm. In September, however, the population profile of AR18 had shifted such that the density was significantly greater in the 0.25- to 0.5 mm size class than in aggregates of <0.25 mm and >1.0 mm. The populations of two other Rhizobium serotypes (AR6 and AS36) followed the same trends of distribution in the June and September samplings. These data indicate the existence of structural microsites that vary in their suitabilities to support growth and protection of bacteria and that are influenced by the presence and type of plant grown in the soil. PMID- 16349533 TI - Comparison of Randomly Amplified Polymorphic DNA with Amplified Fragment Length Polymorphism To Assess Genetic Diversity and Genetic Relatedness within Genospecies III of Pseudomonas syringae. AB - Recently, DNA pairing analyses showed that Pseudomonas syringae pv. tomato and related pathovars, including P. syringae pv. maculicola, form a genomic species (Pseudomonas tomato) (L. Gardan, H. L. Shafik, and P. A. D. Grimont, p. 445-448, in K. Rudolph, T. J. Burr, J. W. Mansfield, D. Stead, A. Vivian, and J. von Kietzell, ed., Pseudomonas syringae Pathovars and Related Pathogens, 1997). The genetic diversity of 23 strains belonging to this genomic species and 4 outgroup strains was analyzed with randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphic (AFLP) techniques. Simple boiling of P. syringae cells was suitable for subsequent DNA amplification to obtain reliable patterns in RAPD and AFLP analyses. In general, the grouping of P. syringae strains by both analysis techniques corresponded well with the classification obtained from an RFLP analysis of ribosomal DNA operons, DNA pairing studies, and an analysis of pathogenicity data. However, two strains of P. syringae pv. maculicola produced distinct DNA patterns compared to the DNA patterns of other P. syringae pv. maculicola strains; these patterns led us to assume that horizontal transfer of DNA could occur between bacterial populations. Both techniques used in this study have high discriminating power because strains of P. syringae pv. tomato and P. syringae pv. maculicola which were indistinguishable by other techniques, including pathogenicity tests on tomato, were separated into two groups by both RAPD and AFLP analyses. In addition, data analysis showed that the AFLP method was more efficient for assessing intrapathovar diversity than RAPD analysis and allowed clear delineation between intraspecific and interspecific genetic distances, suggesting that it could be an alternative to DNA pairing studies. However, it was not possible to distinguish the two races of P. syringae pv. tomato on the basis of an analysis of the data provided by either the AFLP or RAPD technique. PMID- 16349532 TI - Quinone profiling of bacterial communities in natural and synthetic sewage activated sludge for enhanced phosphate removal. AB - Respiratory quinones were used as biomarkers to study bacterial community structures in activated sludge reactors used for enhanced biological phosphate removal (EBPR). We compared the quinone profiles of EBPR sludges and standard sludges, of natural sewage and synthetic sewage, and of plant scale and laboratory scale systems. Ubiquinone (Q) and menaquinone (MK) components were detected in all sludges tested at molar MK/Q ratios of 0.455 to 0.981. The differences in MK/Q ratios were much larger when we compared different wastewater sludges (i.e., raw sewage and synthetic sewage) than when we compared sludges from the EBPR and standard processes or plant scale and laboratory scale systems. In all sludges tested a Q with eight isoprene units (Q-8) was the most abundant quinone. In the MK fraction, either tetrahydrogenated MK-8 or MK-7 was the predominant type, and there was also a significant proportion of MK-6 to MK-8 in most cases. A numerical cluster analysis of the profiles showed that the sludges tested fell into two major clusters; one included all raw sewage sludges, and the other consisted of all synthetic sewage sludges, independent of the operational mode and scale of the reactors and the phosphate accumulation. These data suggested that Q-8-containing species belonging to the class Proteobacteria (i.e., species belonging to the beta subclass) were the major constituents of the bacterial populations in the EBPR sludge, as well as in standard activated sludge. Members of the class Actinobacteria (gram-positive bacteria with high DNA G+C contents) were the second most abundant group in both types of sludge. The bacterial community structures in activated sludge processes may be affected more by the nature of the influent wastewater than by the introduction of an anaerobic stage into the process or by the scale of the reactors. PMID- 16349534 TI - Phase Variation in Xenorhabdus nematophilus. AB - Xenorhabdus nematophilus is a symbiotic bacterium that inhabits the intestine of entomopathogenic nematodes. The bacterium-nematode symbiotic pair is pathogenic for larval-stage insects. The phase I cell type is the form of the bacterium normally associated with the nematode. A variant cell type, referred to as phase II, can form spontaneously under stationary-phase conditions. Phase II cells do not elaborate products normally associated with the phase I cell type. To better define phase variation in X. nematophilus, several strains (19061, AN6, F1, N2-4) of this bacterium were analyzed for new phenotypic traits. An analysis of pathogenicity in Manduca sexta larvae revealed that the phase II form of AN6 (AN6/II) was significantly less virulent than the phase I form (AN6/I). The variant form of N2-4 was also avirulent. On the other hand, F1/II and 19061/II were as virulent as the respective phase I cells. Strain 19061/II was found to be motile, and AN6/II regained motility when the bacteria were grown in low osmolarity medium. In contrast, F1/II remained nonmotile. The phase II cells did not produce the outer membrane protein, OpnB, that is normally induced during the stationary phase. Both phase I and phase II cells were able to support nematode growth and development. These findings indicate that while certain phenotypic traits are common to all phase II cells, other characteristics, such as virulence and motility, are variable and can be influenced by environmental conditions. PMID- 16349535 TI - Analysis of BIOLOG GN Substrate Utilization Patterns by Microbial Communities. AB - BIOLOG GN plates are increasingly used to characterize microbial communities by determining the ability of the communities to oxidize various carbon sources. Studies were done to determine whether the BIOLOG GN plate assay accurately reflects the catabolic potential of the inoculum used. To gain insight into which populations of microbial communities contribute to the BIOLOG patterns, denaturing gradient gel electrophoresis and temperature gradient gel electrophoresis (TGGE) were used to assess the diversity of ribotypes in the inocula and individual wells of BIOLOG plates following incubation. These studies were done with microbial communities from the rhizosphere of potatoes and an activated sludge reactor fed with glucose and peptone. TGGE analyses of BIOLOG wells inoculated with cell suspensions from the potato rhizosphere revealed that, compared with the inoculum, there was a decrease in the number of 16S rRNA gene fragments obtained from various wells, as well as a concomitant loss of populations that had been numerically dominant in the inoculum. The dominant fragments in TGGE gels could be assigned to the gamma subclass of the class Proteobacteria, suggesting that fast-growing bacteria adapted to high substrate concentrations were numerically dominant in the wells and may have been primarily responsible for the patterns of substrate use that were observed. Similarly, the community structure changed in wells inoculated with cells from activated sludge; one or more populations were enriched, but all dominant populations of the inoculum could be detected in at least one well. This study showed that carbon source utilization profiles obtained with BIOLOG GN plates do not necessarily reflect the functional potential of the numerically dominant members of the microbial community used as the inoculum. PMID- 16349536 TI - Development of a laboratory-scale leaching plant for metal extraction from fly ash by thiobacillus strains. AB - Semicontinuous biohydrometallurgical processing of fly ash from municipal waste incineration was performed in a laboratory-scale leaching plant (LSLP) by using a mixed culture of Thiobacillus thiooxidans and Thiobacillus ferrooxidans. The LSLP consisted of three serially connected reaction vessels, reservoirs for a fly ash suspension and a bacterial stock culture, and a vacuum filter unit. The LSLP was operated with an ash concentration of 50 g liter, and the mean residence time was 6 days (2 days in each reaction vessel). The leaching efficiencies (expressed as percentages of the amounts applied) obtained for the economically most interesting metal, Zn, were up to 81%, and the leaching efficiencies for Al were up to 52%. Highly toxic Cd was completely solubilized (100%), and the leaching efficiencies for Cu, Ni, and Cr were 89, 64, and 12%, respectively. The role of T. ferrooxidans in metal mobilization was examined in a series of shake flask experiments. The release of copper present in the fly ash as chalcocite (Cu(2)S) or cuprite (Cu(2)O) was dependent on the metabolic activity of T. ferrooxidans, whereas other metals, such as Al, Cd, Cr, Ni, and Zn, were solubilized by biotically formed sulfuric acid. Chemical leaching with 5 N H(2)SO(4) resulted in significantly increased solubilization only for Zn. The LSLP developed in this study is a promising first step toward a pilot plant with a high capacity to detoxify fly ash for reuse for construction purposes and economical recovery of valuable metals. PMID- 16349537 TI - Spatial and Temporal Deposition of Adhesive Extracellular Polysaccharide Capsule and Fimbriae by Hyphomonas Strain MHS-3. AB - Hyphomonas strain MHS-3, a member of a genus of primary colonizers of surfaces immersed in marine water, synthesizes two structures that mediate adhesion to solid substrata, namely, capsular exopolysaccharide and fimbriae. Specific stains, gold-labelled lectins, and monoclonal antibodies, along with transmission electron microscopy of synchronized populations, revealed that both structures are polarly and temporally expressed. The timed synthesis and placement of the fimbriae and capsule correlated with the timing and locus of MHS-3 adhesion. PMID- 16349538 TI - Effects of Ionic and Osmotic Strength on the Glucosyltransferase of Rhizobium meliloti Responsible for Cyclic beta-(1,2)-Glucan Biosynthesis. AB - The cyclic beta-(1,2)-glucans of Rhizobium meliloti and Agrobacterium tumefaciens play an important role during hypoosmotic adaptation, and the synthesis of these compounds is osmoregulated. Glucosyltransferase, the enzyme responsible for cyclic beta-(1,2)-glucan biosynthesis, is present constitutively, suggesting that osmotic regulation of the biosynthesis of these glucans occurs through modulation of enzyme activity. In this study, we examined regulation of cyclic glucan biosynthesis in vitro with membrane preparations from R. meliloti. The results show that ionic solutes inhibit glucan synthesis, even when they are present at low concentrations (e.g., 10 mM). In contrast, neutral solutes (glucose, sucrose, and the compatible solutes glycine betaine and trehalose) were found to stimulate glucan synthesis in vitro when they were present at high concentrations (e.g., 1 M). Furthermore, high concentrations of these neutral solutes were shown to compensate for the inhibition of glucosyltransferase activity by ionic solutes. Consistent with their ionic character, the compatible solute potassium glutamate and the osmoprotectant choline chloride inhibited glucosyltransferase activity in vitro. The results suggest that intracellular ion concentrations, intracellular osmolarity, and intracellular concentrations of nonionic compatible solutes all act as important determinants of glucosyltransferase activity in vivo. Additional experiments were performed with an ndvA mutant defective for transport of cyclic glucans and an ndvB mutant that produces a C-terminal truncated glucosyltransferase. Cyclic beta-(1,2)-glucan biosynthesis, although reduced, was found to be osmoregulated in both mutants. These results reveal that NdvA and the C terminus of NdvB are not required for osmotic regulation of cyclic beta-(1,2) glucan biosynthesis. PMID- 16349540 TI - Enhancement of Exopolysaccharide Production by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 with a Simplified Defined Medium. AB - The aim of this work was to investigate the medium requirements for growth and production of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772. The strain was grown in batch cultures on a chemically defined medium, and the technique of single omission of medium components was applied to determine the nutritional requirements. The omission of aspartic acid, glutamic acid, or glycine affected growth only slightly, and the omission of glutamine, asparagine, or threonine resulted in a stronger reduction of the growth. All the other amino acids were essential. Multiple omissions of amino acids caused an almost complete loss of growth. L. delbrueckii subsp. bulgaricus required only riboflavin, calcium pantothenate, and nicotinic acid as individual vitamins. Surprisingly, when only these vitamins were present in the medium and other vitamins were not, less growth was observed than in the complete medium but the amount of exopolysaccharide produced was significantly greater. These observations were studied in more detail with a simplified defined medium in which L. delbrueckii subsp. bulgaricus was able to grow and produce exopolysaccharides. Although the final optical density in the simplified medium was lower, the production of exopolysaccharides was about twofold higher than in the complete medium. PMID- 16349539 TI - Characterization of the Different Dextransucrase Activities Excreted in Glucose, Fructose, or Sucrose Medium by Leuconostoc mesenteroides NRRL B-1299. AB - When grown in glucose or fructose medium in the absence of sucrose, Leuconostoc mesenteroides NRRL B-1299 produces two distinct extracellular dextransucrases named glucose glucosyltransferase (GGT) and fructose glucosyltransferase (FGT). The production level of GGT and FGT is 10 to 20 times lower than that of the extracellular dextransucrase sucrose glucosyltransferase (SGT) produced on sucrose medium (traditional culture conditions). GGT and FGT were concentrated by ultrafiltration before sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Their molecular masses were 183 and 186 kDa, respectively, differing from the 195 kDa of SGT. The structural analysis of the dextran produced from sucrose and of the oligosaccharides synthesized by acceptor reaction in the presence of maltose showed that GGT and FGT are two different enzymes not previously described for this strain. The polymer synthesized by GGT contains 30% alpha(1-->2) linkages, while FGT catalyzes the synthesis of a linear dextran only composed of alpha(1-->6) linkages. PMID- 16349541 TI - Effects of Soil on Ammonia, Ethylene, Chloroethane, and 1,1,1-Trichloroethane Oxidation by Nitrosomonas europaea. AB - Ammonia monooxygenase (AMO) from Nitrosomonas europaea catalyzes the oxidation of ammonia to hydroxylamine and has been shown to oxidize a variety of halogenated and nonhalogenated hydrocarbons. As part of a program focused upon extending these observations to natural systems, a study was conducted to examine the influence of soil upon the cooxidative abilities of N. europaea. Small quantities of Willamette silt loam (organic carbon content, 1.8%; cation-exchange capacity, 15 cmol/kg of soil) were suspended with N. europaea cells in a soil-slurry-type reaction mixture. The oxidations of ammonia and three different hydrocarbons (ethylene, chloroethane, and 1,1,1-trichloroethane) were compared to results for controls in which no soil was added. The soil significantly inhibited nitrite production from 10 mM ammonium by N. europaea. Inhibition resulted from a combination of ammonium adsorption onto soil colloids and the exchangeable acidity of the soil lowering the pH of the reaction mixture. These phenomena resulted in a substantial drop in the concentration of NH(4) in solution (10 to 4.5 mM) and, depending upon the pH, in a reduction in the amount of available NH(3) to concentrations (8 to 80 muM) similar to the K(s) value of AMO for NH(3) ( approximately 29 muM). At a fixed initial pH (7.8), the presence of soil also modified the rates of oxidation of ethylene and chloroethane and changed the concentrations at which their maximal rates of oxidation occurred. The modifying effects of soil on nitrite production and on the cooxidation of ethylene and chloroethane could be circumvented by raising the ammonium concentration in the reaction mixture from 10 to 50 mM. Soil had virtually no effect on the oxidation of 1,1,1-trichloroethane. PMID- 16349542 TI - Effect of Temperature on Adhesion of Vibrio Strain AK-1 to Oculina patagonica and on Coral Bleaching. AB - Laboratory aquarium experiments demonstrated that Vibrio strain AK-1 caused rapid and extensive bleaching of the coral Oculina patagonica at 29 degrees C, slower and less-complete bleaching at 23 degrees C, and no bleaching at 16 degrees C. At 29 degrees C, the application of approximately 100 Vibrio strain AK-1 cells directly onto the coral caused 50 and 83% bleaching after 10 and 20 days, respectively. At 16 degrees C, there was no bleaching, even with an initial inoculum of 1.2 x 10 bacteria. To begin to understand the effect of seawater temperature on bleaching of O. patagonica by Vibrio strain AK-1, adhesion of the bacteria to the coral as a function of temperature was studied. Inoculation of 10Vibrio strain AK-1 organisms into flasks containing 20 ml of seawater at 25 degrees C and a fragment of O. patagonica resulted in net levels of bacterial adhesion to the coral of 45, 78, and 84% after 2, 6, and 8 h, respectively. The adhesion was inhibited 65% by 0.001% d-galactose and 94% by 0.001% methyl-beta-d galactopyranoside (beta-M-Gal). After the incubation of Vibrio strain AK-1 with the coral for 6 h, 42% of the input bacteria were released from the coral with 0.01% beta-M-Gal, compared to less than 0.2% when beta-M-Gal was present during the adhesion step. Adhesion did not occur when Vibrio strain AK-1 was grown at 16 degrees C, regardless of whether the corals were maintained at 16 or 25 degrees C, whereas bacteria grown at 25 degrees C adhered to corals maintained at 16 or 25 degrees C. Bacteria grown at 25 degrees C adhered avidly to Sepharose beads containing covalently bound beta-d-galactopyranoside but failed to bind if grown at 16 degrees C. These data suggest that elevated seawater temperatures may cause coral bleaching by allowing for the expression of adhesin genes of Vibrio strain AK-1. PMID- 16349543 TI - Synergism between Bacillus thuringiensis Spores and Toxins against Resistant and Susceptible Diamondback Moths (Plutella xylostella). AB - We studied the effects of combinations of Bacillus thuringiensis spores and toxins on the mortality of diamondback moth (Plutella xylostella) larvae in leaf residue bioassays. Spores of B. thuringiensis subsp. kurstaki increased the toxicity of crystals of B. thuringiensis subsp. kurstaki to both resistant and susceptible larvae. For B. thuringiensis subsp. kurstaki, resistance ratios were 1,200 for a spore-crystal mixture and 56,000 for crystals without spores. Treatment of a spore-crystal formulation of B. thuringiensis subsp. kurstaki with the antibiotic streptomycin to inhibit spore germination reduced toxicity to resistant larvae but not to susceptible larvae. In contrast, analogous experiments with B. thuringiensis subsp. aizawai revealed no significant effects of adding spores to crystals or of treating a spore-crystal formulation with streptomycin. Synergism occurred between Cry2A and B. thuringiensis subsp. kurstaki spores against susceptible larvae and between Cry1C and B. thuringiensis subsp. aizawai spores against resistant and susceptible larvae. The results show that B. thuringiensis toxins combined with spores can be toxic even though the toxins and spores have little or no independent toxicity. Results reported here and previously suggest that, for diamondback moth larvae, the extent of synergism between spores and toxins of B. thuringiensis depends on the strain of insect, the type of spore, the set of toxins, the presence of other materials such as formulation ingredients, and the concentrations of spores and toxins. PMID- 16349544 TI - Differential Effects of Dimethylsulfoniopropionate, Dimethylsulfonioacetate, and Other S-Methylated Compounds on the Growth of Sinorhizobium meliloti at Low and High Osmolarities. AB - An extract from the marine alga Ulva lactuca was highly osmoprotective in salt stressed cultures of Sinorhizobium meliloti 102F34. This beneficial activity was due to algal 3-dimethylsulfoniopropionate (DMSP), which was accumulated as a dominant compatible solute and strongly reduced the accumulation of endogenous osmolytes in stressed cells. Synthetic DMSP also acted as a powerful osmoprotectant and was accumulated as a nonmetabolizable cytosolic osmolyte (up to a concentration of 1,400 nmol/mg of protein) throughout the growth cycles of the stressed cultures. In contrast, 2-dimethylsulfonioacetate (DMSA), the sulfonium analog of the universal osmoprotectant glycine betaine (GB), was highly toxic to unstressed cells and was not osmoprotective in stressed cells of wild type strains of S. meliloti. Nonetheless, the transport and accumulation of DMSA, like the transport and accumulation of DMSP and GB, were osmoregulated and increased fourfold in stressed cells of strain 102F34. Strikingly, DMSA was not toxic and became highly osmoprotective in mutants that are impaired in their ability to demethylate GB and DMSA. Furthermore, 2-methylthioacetate and thioglycolic acid (TGA), the demethylation products of DMSA, were excreted, apparently as a mechanism of cellular detoxification. Also, exogenous TGA and DMSA displayed similar inhibitory effects in strain 102F34. Thus, on the basis of these findings and other physiological and biochemical evidence, we infer that the toxicity of DMSA in wild-type strains of S. meliloti stems from its catabolism via the GB demethylation pathway. This is the first report describing the toxicity of DMSA in any organism and a metabolically stable osmoprotectant (DMSP) in S. meliloti. PMID- 16349545 TI - Detection of infectious tobamoviruses in forest soils. AB - Our objectives were to evaluate elution and bait plant methods to detect infectious tobamoviruses in forest soils in New York State. Soils were collected from two forest sites: Whiteface Mountain (WF) and Heiberg Forest (HF). The effectiveness of four buffers to elute tomato mosaic tobamovirus (ToMV) from organic and mineral fractions of WF soil amended with ToMV was tested, and virus content was assessed by enzyme-linked immunosorbent assay (ELISA). The effectiveness of Chenopodium quinoa (Willd.) bait plants to detect the virus also was tested. Both methods then were utilized to detect tobamoviruses in 11 WF and 2 HF soil samples. A phosphate buffer (100 mM, pH 7.0) eluted more ToMV from soil than the other buffers tested. Mineral soil bound more virus than organic soil. Virus recoveries from virus-amended organic and mineral soils were 3 and 10%, respectively, and the detection sensitivity was 10 to 20 ng/g of soil. Roots of bait plants grown in all virus-amended soils tested positive by ELISA, and virus concentrations averaged 10 ng/g. Both ToMV and tobacco mosaic tobamovirus (TMV) were transmitted to C. quinoa by elution from one of two HF soil samples but not from the WF soil samples. A tobamovirus was detected by bait planting in 12 of 73 (16%) root extracts representing 5 of 13 soil samples (38%). Tobamovirus-like particles were seen by transmission electron microscopy in 6 of 12 infected root extracts. Tobamoviruses occur in forest soils in New York State. Abiotic soil transmission to trees may permit localized spread and persistence of these viruses in forest ecosystems. PMID- 16349546 TI - Seasonal pattern of tomato mosaic tobamovirus infection and concentration in red spruce seedlings. AB - Tomato mosaic tobamovirus (ToMV) infects red spruce (Picea rubens) and causes significant changes in its growth and physiology. The mechanism of infection and the pattern of virus concentration in seedling roots and needles were investigated. One-year-old red spruce seedlings were obtained from the nursery in April and June 1995 and August 1996 and tested for ToMV using enzyme-linked immunosorbent assay (ELISA). Virus-free seedlings were divided into three treatments: control, root inoculated, and needle inoculated. Two control, five root-inoculated, and five needle-inoculated seedlings were sampled destructively at biweekly intervals for 3 months and then tested for ToMV by ELISA. ToMV was transmitted to seedlings by root but not by needle inoculation. The virus was detected in 67 to 100% of roots but in less than 7% of needles of root-inoculated seedlings. The percent infection of root-inoculated seedlings differed significantly between the April and June and between the April and August inoculation periods. Virus concentration in infected seedling roots increased initially, peaked within 4 weeks postinoculation, and steadily declined thereafter. Significant differences in ToMV concentrations in roots also were detected among inoculation periods and sampling dates. Early spring may represent the optimal time for infection of seedlings, as well as for assaying roots for ToMV. PMID- 16349547 TI - Analysis of the beta-1,3-Glucanolytic System of the Biocontrol Agent Trichoderma harzianum. AB - The biocontrol agent Trichoderma harzianum IMI206040 secretes beta-1,3-glucanases in the presence of different glucose polymers and fungal cell walls. The level of beta-1,3-glucanase activity secreted was found to be proportional to the amount of glucan present in the inducer. The fungus produces at least seven extracellular beta-1,3-glucanases upon induction with laminarin, a soluble beta 1,3-glucan. The molecular weights of five of these enzymes fall in the range from 60,000 to 80,000, and their pIs are 5.0 to 6.8. In addition, a 35-kDa protein with a pI of 5.5 and a 39-kDa protein are also secreted. Glucose appears to inhibit the formation of all of the inducible beta-1,3-glucanases detected. A 77 kDa glucanase was partially purified from the laminarin culture filtrate. This enzyme is glycosylated and belongs to the exo-beta-1,3-glucanase group. The properties of this complex group of enzymes suggest that the enzymes might play different roles in host cell wall lysis during mycoparasitism. PMID- 16349548 TI - Release of dimethylsulfide from dimethylsulfoniopropionate by plant-associated salt marsh fungi. AB - The range of types of microbes with dimethylsulfoniopropionate (DMSP) lyase capability (enzymatic release of dimethylsulfide [DMS] from DMSP) has recently been expanded from bacteria and eukaryotic algae to include fungi (a species of the genus Fusarium [M. K. Bacic and D. C. Yoch, Appl. Environ. Microbiol. 64:106 111, 1998]). Fungi (especially ascomycetes) are the predominant decomposers of shoots of smooth cordgrass, the principal grass of Atlantic salt marshes of the United States. Since the high rates of release of DMS from smooth cordgrass marshes have a temporal peak that coincides with peak shoot death, we hypothesized that cordgrass fungi were involved in this DMS release. We tested seven species of the known smooth cordgrass ascomycetes and discovered that six of them exhibited DMSP lyase activity. We also tested two species of ascomycetes from other DMSP-containing plants, and both were DMSP lyase competent. For comparison, we tested 11 species of ascomycetes and mitosporic fungi from halophytes that do not contain DMSP; of these 11, only 3 were positive for DMSP lyase. A third group tested, marine oomycotes (four species of the genera Halophytophthora and Pythium, mostly from mangroves), showed no DMSP lyase activity. Two of the strains of fungi found to be positive for DMSP lyase also exhibited uptake of DMS, an apparently rare combination of capabilities. In conclusion, a strong correlation exists between a fungal decomposer's ability to catabolize DMSP via the DMSP lyase pathway and the host plant's production of DMSP as a secondary product. PMID- 16349549 TI - Evidence that a New Antibiotic Flavone Glycoside Chemically Defends the Sea Grass Thalassia testudinum against Zoosporic Fungi. AB - Significantly fewer thraustochytrid protists (zoosporic fungi) were observed in association with healthy leaf tissue of the marine angiosperm Thalassia testudinum than in association with sterilized samples that were returned to the collection site for 48 h. In support of the hypothesis that sea grass secondary metabolites were responsible for these differences, extracts of healthy T. testudinum leaf tissues inhibited the growth of the co-occurring thraustochytrid Schizochytrium aggregatum and deterred the attachment of S. aggregatum motile zoospores to an extract-impregnated substrate. By using S. aggregatum for bioassay-guided chemical fractionation, a new flavone glycoside was isolated and structurally characterized as luteolin 7-O-beta-d-glucopyranosyl-2"-sulfate. Whole-leaf tissue concentrations of this metabolite (4 mg/ml of wet leaf tissue) inhibited S. aggregatum attachment, and a significantly lower concentration (270 mug/ml) reduced thraustochytrid growth by 50%, suggesting that natural concentrations are at least 15 times greater than that needed for significant microbiological effects. These results offer the first complete chemical characterization of a sea grass sulfated flavone glycoside and provide evidence that a secondary metabolite chemically defends T. testudinum against fouling microorganisms. PMID- 16349550 TI - Ammonium Limitation Results in the Loss of Ammonia-Oxidizing Activity in Nitrosomonas europaea. AB - The effects of limiting concentrations of ammonium on the metabolic activity of Nitrosomonas europaea, an obligate ammonia-oxidizing soil bacterium, were investigated. Cells were harvested during late logarithmic growth and were incubated for 24 h in growth medium containing 0, 15, or 50 mM ammonium. The changes in nitrite production and the rates of ammonia- and hydroxylamine dependent oxygen consumption were monitored. In incubations without ammonium, there was little change in the ammonia oxidation activity after 24 h. With 15 mM ammonium, an amount that was completely consumed, there was an 85% loss of the ammonia oxidation activity after 24 h. In contrast, there was only a 35% loss of the ammonia oxidation activity after 24 h in the presence of 50 mM ammonium, an amount that was not consumed to completion. There was little effect on the hydroxylamine oxidation activity in any of the incubations. The loss of ammonia oxidation activity was not due to differences in steady-state levels of ammonia monooxygenase (AMO) mRNA (amoA) or to degradation of the active site-containing subunit of AMO protein. The incubations were also conducted at a range of pH values to determine whether the loss of ammonia oxidation activity was correlated to the residual ammonium concentration. The loss of ammonia oxidation activity after 24 h was less at lower pH values (where the unoxidized ammonium concentration was higher). When added in conjunction with limiting ammonium, short-chain alkanes, which are alternative substrates for AMO, prevented the loss of ammonia oxidation activity at levels corresponding to their binding affinity for AMO. These results suggest that substrates of AMO can preserve the ammonia oxidizing activity of N. europaea in batch incubations by protecting either AMO itself or other molecules associated with ammonia oxidation. PMID- 16349551 TI - Temperature and sporulation of aquatic hyphomycetes. AB - Temperature appears to be an important factor affecting the occurrence and distribution of aquatic hyphomycetes, the dominant leaf litter-decomposing fungi in streams. We compared conidium production by eight species of aquatic hyphomycetes grown on yellow poplar leaves in stream-simulating microcosms at three temperatures (15, 20, and 25 degrees C). The greatest conidium production occurred at 15 degrees C for one species, 20 degrees C for two species, and 25 degrees C for two species. Two species produced similar numbers of conidia at 20 and 25 degrees C, and one species produced similar numbers of conidia at all three temperatures. Linear growth rates were determined on malt extract agar. Six species had the same pattern of temperature responses for growth on malt extract agar as for sporulation on leaves, as shown by the positive correlations between the two parameters at the three temperatures. The species examined also exhibited differences in number of conidia produced from a similar amount of leaf material at a given temperature. These differences appeared to be due primarily to differences in individual conidium mass (determined by weighing conidia produced from cultures), as shown by the relationship of the type Y = k/X (r = 0.96), where Y is the number of conidia produced, X is the individual conidium mass in milligrams, and k is a constant empirically determined to be 2.11. This finding supports the hypothesis that aquatic hyphomycetes allocate similar amounts of their resources to reproduction but vary with respect how these resources are partitioned into reproductive units (conidia). PMID- 16349552 TI - Effects of Cellular Metabolism and Viability on Metal Ion Accumulation by Cultured Biomass from a Bloom of the Cyanobacterium Microcystis aeruginosa. AB - The sorption of nickel, cadmium, and copper by cultured biomass from a naturally occurring bloom of Microcystis aeruginosa was demonstrated in two systems: cells suspended in culture medium and cells immobilized in alginate. Incubation in the absence of light, in the presence of metabolic inhibitors, and at 4 degrees C did not substantially decrease the copper accumulation by cells in culture medium. Heat-killed, formaldehyde-treated, and air-dried biomass samples sorbed nearly as much (or in some cases slightly more) copper as did viable samples. PMID- 16349553 TI - A small-scale method for screening of lignin-degrading microorganisms. AB - A new method to facilitate rapid screening of lignin-degrading microorganisms was developed. Fungal strains are cultivated in tissue culture plates containing C ring-labeled dehydrogenation polymerizate (DHP) (synthetic lignin). Evolved CO(2) is trapped in barium-saturated filter paper and is detected by exposing the paper to X-ray film. Analysis of the autoradiograms, carried out by density measurement with an image analysis program, allows for a semiquantitative estimation of the amount of CO(2) evolved. The method is especially useful for screening for new, powerful lignin-degrading strains in both man-made and natural environments. It eliminates the need for special equipment for their cultivation and trapping of CO(2) as well as laborious sample analysis. The method has in this study been used to test three novel fungal isolates and a laccaseless mutant of the basidiomycete Pycnoporus cinnabarinus. Their ligninolytic capacities were compared with those of the potent lignin degrader Ceriporiopsis subvermispora. PMID- 16349554 TI - Anaerobic Oxidation of [1,2-C]Dichloroethene under Mn(IV)-Reducing Conditions. AB - Anaerobic oxidation of [1,2-C]dichloroethene to CO(2) under Mn(IV)-reducing conditions was demonstrated. The results indicate that oxidative degradation of partially chlorinated solvents like dichloroethene can be significant even under anoxic conditions and demonstrate the potential importance of Mn(IV) reduction for remediation of chlorinated groundwater contaminants. PMID- 16349555 TI - Changes in Permeability of Brush Border Membrane Vesicles from Spodoptera littoralis Midgut Induced by Insecticidal Crystal Proteins from Bacillus thuringiensis. AB - Bacillus thuringiensis insecticidal crystal proteins (ICPs) are thought to induce pore formation in midgut cell membranes of susceptible insects. Cry1Ca, which is significantly active in Spodoptera littoralis, made brush border membrane vesicles permeable to KCl (osmotic swelling was monitored by the light scattering technique); the marginally active ICPs Cry1Aa, Cry1Ab, and Cry1Ac did not. PMID- 16349557 TI - Anaerobic Degradation of Hexadecan-2-one by a Microbial Enrichment Culture under Sulfate-Reducing Conditions. AB - A microbial enrichment culture from marine sediment was able to grow on hexadecan 2-one as the sole source of carbon and energy under sulfate-reducing conditions. Oxidation of the ketone involved carboxylation reactions and was coupled to sulfide production. This enrichment culture also grew on 6,10,14 trimethylpentadecan-2-one. PMID- 16349556 TI - Laser microsurgery permits fungal plasma membrane single-ion-channel resolution at the hyphal tip. AB - A method for formation of high-electrical-resistance seals on the Neurospora crassa plasma membrane, allowing resolution of single-ion-channel activity by patch clamp electrophysiology, is reported. Laser microsurgery permits access to the hyphal apex without enzymatic cell wall digestion and loss of morphological polarity. Cell wall reformation is delayed by brefeldin. This method can allow full characterization of apical plasma membrane channels, which are implicated in tip growth. PMID- 16349561 TI - Dibiphytanyl ether lipids in nonthermophilic crenarchaeotes. AB - [This corrects the article on p. 1136 in vol. 64.]. PMID- 16349564 TI - Evidence that a New Antibiotic Flavone Glycoside Chemically Defends the Sea Grass Thalassia testudinum against Zoosporic Fungi. AB - [This corrects the article on p. 1491 in vol. 64.]. PMID- 16349569 TI - Confirmational Identification of Escherichia coli, a Comparison of Genotypic and Phenotypic Assays for Glutamate Decarboxylase and beta-d-Glucuronidase. AB - [This corrects the article on p. 3350 in vol. 62, PMID: 8795225.]. PMID- 16349570 TI - Sensitive and Rapid Detection of Viable Giardia Cysts and Cryptosporidium parvum Oocysts in Large-Volume Water Samples with Wound Fiberglass Cartridge Filters and Reverse Transcription-PCR. AB - [This corrects the article on p. 1743 in vol. 64, PMID: 9572946.]. PMID- 16349571 TI - New Nitrogen-Fixing Microorganisms Detected in Oligotrophic Oceans by Amplification of Nitrogenase (nifH) Genes. AB - [This corrects the article on p. 3444 in vol. 64, PMID: 9726895.]. PMID- 16349573 TI - Metabolic Behavior and Chlortetracycline Production by Streptomyces aureofaciens in Liquid Culture. PMID- 16349572 TI - The Microbiological Deterioration of Rubber Insulation. PMID- 16349574 TI - Specific Studies on the Soil-Block Procedure for Bioassay of Wood Preservatives. PMID- 16349575 TI - Movement of Certain Antibiotics in Cuttings of Pyracantha and Carnation. PMID- 16349576 TI - Newer Preservative Treatments for Wood. PMID- 16349577 TI - Acid Production by Homofermentative Lactobacilli at Controlled pH as a Tool for Studying the Unit Process of Fermentation. PMID- 16349578 TI - Some Auxiliary Effects of Textile Fungicides. PMID- 16349579 TI - A Review of the Comparative Bacteriology of Hevea Latex and Its Commercial Derivatives. PMID- 16349580 TI - Cellulose Decomposition in Soil Burial Beds: I. Soil Properties in Relation to Cellulose Degradation. PMID- 16349581 TI - Cellulose Decomposition in Soil Burial Beds: II. Cellulolytic Activity as Influenced by Alteration of Soil Properties. PMID- 16349582 TI - A Laboratory Screening Test of Bactericides for Use in Sheep Pelt Soaking Pits. PMID- 16349583 TI - Aerobic Bacteria Involved in the Retting of Jute. PMID- 16349584 TI - Pectin Decomposition by Species of Pseudomonas and Their Role in the Retting of Malvaceous Plants. PMID- 16349585 TI - Studies in Feed Spoilage: Inhibition of Mold Growth by Gamma Radiation. PMID- 16349586 TI - Chemistry and Microbiology of Forage-Crop Silage. PMID- 16349587 TI - Factors Involved in the Control of Microbial Deterioration: I. Variation in Sensitivity of Different Strains of the Same Species. PMID- 16349588 TI - Comparative Incidence of Coliform Bacteria and Enterococci in Undisturbed Soil. PMID- 16349589 TI - Separation of Microorganisms by Flotation: I. Development and Evaluation of Assay Procedures. PMID- 16349590 TI - Separation of Microorganisms by Flotation: II. Flotation of Spores of Bacillus subtilis var. niger. PMID- 16349591 TI - Some Problems Involved in the Use of Quaternary Ammonium Compounds as Fabric Disinfectants. PMID- 16349592 TI - Changes in the Paper Electrophoretic Protein Patterns of Refrigerated Skim Milk Accompanying Growth of Three Pseudomonas Species. PMID- 16349593 TI - Effect of Antibiotic and Chemical Dips on the Microflora of Packaged Salad Mix. PMID- 16349594 TI - Antifungal Activity of Some Thiosemicarbazones and Their Copper Complexes. PMID- 16349595 TI - Biosynthesis of Lysine from alpha-Ketoadipic Acid and alpha-Aminoadipic Acid in Yeast. PMID- 16349596 TI - Influence of Subtle Differences in Plating Procedure on Bacterial Counts of Prepared Frozen Foods. PMID- 16349597 TI - A Plaque Suppression Method for the Study of Antiviral Compounds. PMID- 16349599 TI - An Approach to the Kinetics of Microbiological Deterioration. PMID- 16349598 TI - Comparative Study of Penicillin Production with Vegetative and Spore Inoculum of Penicillium chrysogenum. PMID- 16349600 TI - Continuous Culture as a Method for Studying Rumen Fermentation. PMID- 16349601 TI - A Study of Bacterial Species from the Rumen Which Produce Ammonia from Protein Hydrolyzate. PMID- 16349602 TI - Some Nutritional Requirements of the Genus Ruminococcus. PMID- 16349603 TI - Studies on the Nitrogen Requirements of Some Ruminal Cellulolytic Bacteria. PMID- 16349604 TI - Preliminary Observations on the Effect of Sodium Alginate on Selected Nonsporing Organisms. PMID- 16349605 TI - Growth of Streptomyces aureofaciens in Continuous Culture. PMID- 16349607 TI - Studies on Media for Enumerating Enterococci in Frozen Vegetables. AB - A study was made of the relative sensitivity and specificity of presumptive and confirmatory media for the most probable number enumeration of enterococci in frozen vegetables. Azide dextrose broth yielded the highest numbers of confirmable enterococci and its sensitivity was shown to be comparable to nonselective media. The use of ethyl violet azide broth as a confirmatory medium resulted in a significant number of false positive tests. Growth in broth containing 6.5% sodium chloride incubated at 45 C for 2 days was found to be a more specific confirmatory test for enterococci. PMID- 16349606 TI - Preservation of Microorganisms by Freeze-drying: II. The Destructive Action of Oxygen. Additional Stabilizers for Serratia marcescens. Experiments with Other Microorganisms. PMID- 16349608 TI - Reaction of the Sterilant, Ethylene Oxide, on Plastics. PMID- 16349609 TI - The Occurrence of Gallionella in Salt Water. AB - A Gallionella sp. is described that lives and freely multiplies in sea water. The habitat of the organism has been thoroughly investigated and the possibility of accidental occurrence has been eliminated. The appearance of the Gallionella sp. described is quite different from the similar iron bacteria occurring in fresh water. PMID- 16349610 TI - Bacterial Oxidation of Pyritic Materials in Coal. AB - Applicability of the manometric method for studying the oxidation of pyritic material in the presence of bacteria has been demonstrated. Resting cells of Ferrobacillus ferrooxidans accelerated the oxidation of coal pyrites and coarsely crystalline marcasite, but were inactive on coarsely crystalline pyrite. Resting cells of Thiobacillus thiooxidans were inactive on all pyrites tested. Oxidation rates in the presence of Ferrobacillus were increased by reducing the particle size of pyritic samples, and, in one case, by removing the CaCO(3) from a calcite containing sample. PMID- 16349611 TI - Oxytetracycline Formation by Streptomyces rimosus in Chemically Defined Media. AB - Details in the fermentation of oxytetracycline in a synthetic medium with Streptomyces rimosus have been presented. In these studies, an organic nitrogen source was shown to be essential for the production of significant amounts of antibiotic activity. Of the amino acids tested, aspartic acid, proline, threonine, valine, and beta-alanine were utilized well for both growth and antibiotic production. Markedly different fermentation patterns were observed with aspartic acid and beta-alanine. Glycerol and glucose supported antibiotic yields superior to those found with other carbohydrates tested. Short chain organic acids were not effectively utilized for growth in the absence of a readily fermentable carbohydrate. PMID- 16349612 TI - Occurrence of Enterococci: Bud, Blossom, and Soil Studies. AB - The occurrence of enterococci (group D streptococci) on buds and flowers of plants and in soils has been studied. They were recovered from 27.5% of the flowers of seven species of plants, and from 6.8% of the buds of the same plants. They were recovered from 34% of the flowers of nonagricultural plants, from 32.2% of the flowers of ten species of agricultural dicotyledonous plants, and from 10.4% of the flowers of five species of grasses and cereals. The enterococci were invariably present or invariably absent from all samples taken from very few species. They occurred in small numbers on enclosed tassels and silks of corn of 22 of 60 samples, and in greater numbers on 90% or more of these after their floral parts had emerged. Interposition of a mechanical barrier reduced the incidence of recovery from flowers. The occurrence in soil, generally at a low level of population, may be correlated with occurrence on the plant growing on the soil or with nearby enterococcal-bearing plants. It is concluded that enterococci may be regarded as temporary residents on plants, capable of limited reproduction, and that they are disseminated among plants by the action of insects and wind, and spread to the ground by these agencies, gravity, and rain. PMID- 16349613 TI - A Method for the Determination of Antimicrobial Properties of Treated Fabrics. AB - A method has been presented for evaluating the effectiveness of fabrics treated with antimicrobial agents to resist growth of contaminating organisms. The method consists of direct inoculation of the fabric with suitable test organisms and the subsequent enumeration of growing colonies which develop when the fabric is implanted in a nutrient medium. The method is sensitive, versatile, easy to perform, highly reproducible, and realistically evaluates the practical effectiveness of residual antimicrobial agents on fabrics. Data are given which demonstrate the sensitivity of the method to small differences in antimicrobial concentrations. The versatility of the method is shown by the variety of antimicrobial agents and organisms which have been utilized. Both bacteriostatic and bactericidal activity can be evaluated. PMID- 16349615 TI - "Rifomycin. XIV. Production of Rifomycin B". PMID- 16349614 TI - Production of Fumaric Acid in 20-Liter Fermentors. AB - The conditions necessary for the production of fumaric acid in 20-liter fermentors by fermentation of glucose with Rhizopus arrhizus strain NRRL 2582 were determined. Continuous neutralization of fumaric acid was necessary for optimal yields. Yields of the calcium salt were in excess of 65 g of fumaric acid from 100 g of sugar consumed during fermentation of sugar concentrations of 10 to 16%. Conditions established for calcium fumarate production include a simple mineral salts medium, 0.5 v:v:min aeration rate, 300 rev/min agitation rate in a baffled tank, 33 C incubation temperature, CaCO(3) to neutralize the acid formed, and a 4 to 5% (v/v) vegetative inoculum. A suitable procedure and medium for the preparation of a vigorous vegetative inoculum were established. The tendency for calcium fumarate fermentations to foam excessively was controlled with a proper antifoam agent added prior to sterilization of the medium and again at daily intervals during fermentation. The production of soluble sodium or potassium fumarates was inhibited when the concentration of fumarates reached 3.5 to 4.0%. No means of overcoming this inhibition was found. Starches and certain other grain-derived carbohydrates were fermented to form calcium fumarate in flask experiments with approximately the same efficiency as was glucose. PMID- 16349616 TI - Isolation of Pure Cultures of Algae from Contaminated Cultures. AB - Algal cultures have been freed of bacterial contamination by a procedure involving treatment with a detergent and phenol, followed by plating on an agar medium for selection of pure cultures. PMID- 16349617 TI - Studies on Induction and Repression in Activated Sludge Systems. AB - Both repression and induction of substrate utilization have been the subject of many basic research investigations employing pure cultures. In this investigation these effects were studied using heterogeneous microbial populations prevalent in such biological treatment processes as activated sludge systems. Diauxic substrate removal by activated sludge was observed in a multicomponent medium consisting of glucose and sorbitol. The sludge was acclimated solely to sorbitol; however, the presence of glucose blocked sorbitol removal until glucose was completely utilized. Both diphasic and triphasic oxygen utilization was shown for activated sludges metabolizing multicomponent synthetic wastes consisting of glucose, melibiose, and lactose. It appears from these studies that melibiose utilization was suppressed by the presence of glucose and, although melibiose induced acclimation to lactose, the presence of melibose suppressed lactose utilization. Studies were also conducted using glycogen and starch systems in which it was found that acclimation to either compound conferred immediate acclimation to the other. It was also found that loss of acclimation to lactose was a passive phenomenon and its kinetics could be predicted on the basis of simple diluting out of the enzyme(s) responsible for such acclimation. PMID- 16349618 TI - Growth-Biosynthesis Relationships in Erythromycin Fermentation. AB - The relationship of erythromycin biosynthesis to growth appears to vary with the fermentation medium in which the microorganism is grown. In a medium containing a complex nitrogen source, antibiotic production occurred only when growth was halted, whereas in a glycine-containing synthetic medium the two parameters increased at nearly the same time. An autostimulation of antibiotic production through addition of erythromycin is reported. Also described is the use of a labeled precursor as an index of biosynthesis rates during short periods of time. PMID- 16349619 TI - Culture and Metabolism of the Rumen Ciliate Epidinium ecaudatum Crawley. AB - The rumen ciliate Epidinium ecaudatum was cultured for 6 months in the presence of two strains of bacteria, starch, alfalfa, linseed oil meal, and buffered saline. The cultures required daily transfer and addition of fresh substrate. The protozoan degraded starch, soybean oil meal, linseed oil meal, and cottonseed oil meal, and the fermentation end products from the breakdown of starch were acetic and butyric acids with traces of formic, propionic, and lactic acids, carbon dioxide, and hydrogen. The relationship of E. ecaudatum to other species of rumen oligotrichs and bacteria is discussed. PMID- 16349620 TI - Successive Microbial Populations in Calimyrna Figs. AB - Smyrna-type (Calimyrna) figs have essentially sterile internal tissue until visited by the pollinating fig wasp, Blastophaga psenes, which introduces a specific microflora consisting of Candida guilliermondii var. carpophila and Serratia plymuthica. This flora persists and develops in numbers throughout the ripening period until maturity of the fruit. These organisms do not cause spoilage. The presence of C. guilliermondii var. carpophila appears to increase the attractiveness of the fruit to drosophilae. Drosophila (mainly D. melanogaster) carry spoilage yeasts and bacteria on their exterior body parts, and introduce these organisms during ovipositing in the fruit cavity. The spoilage yeasts consist almost entirely of apiculate yeasts (Hanseniaspora valbyensis, H. uvarum, and Kloeckera apiculata) and of Torulopsis stellata, which cause active fermentative spoilage. Spoilage bacteria (primarily Acetobacter melanogenus) are also introduced with the yeasts. Organic acids are produced by these yeasts as well as by the Acetobacter. A number of minor spoilage yeasts were also identified. PMID- 16349621 TI - Formation of Extracellular Sphingolipids by Microorganisms: IV. Pilot-Plant Production of Tetraacetylphytosphingosine by Hansenula ciferrii. AB - Tetraacetylphytosphingosine (TAPS) formation by the F-60-10 mating type strain of the yeast Hansenula ciferrii, previously observed on agar plates, has been shown to take place in submerged cultures. The optimal conditions for TAPS formation, and the correlation of TAPS production and sugar utilization under aerobic conditions, were studied in 10-liter fermentors. For each gram of glucose consumed, 5 mg of TAPS were formed; for each gram of yeast solids produced, 15 mg of TAPS were synthesized. A 750-liter pilot-plant run yielded 175 g of crude TAPS, which were obtained by hexane extraction of centrifuged yeast cells. PMID- 16349622 TI - Quantitative Studies of Total Lipids of Pathogenic Fungi. AB - Ten species of dermatophytes and four of the systemic fungi were assayed for total lipids, acetone-soluble fraction, and phospholipid content in different types of cultures. The yeast phase of each of the systemic fungi grown on solid medium exhibited a higher total lipid content than did the mycelial growth in liquid medium, either shake or still. Shake cultures, in all the fungi tested, produced the least lipids. The yeasts were consistently higher also in the acetone-soluble fraction. Histoplasma duboisii in the yeast phase and Microsporum gypseum produced the greatest amount of phospholipid, and Blastomyces dermatitidis in the yeast phase and M. canis produced the largest acetone-soluble fraction among the systemic fungi and dermatophytes, respectively. PMID- 16349623 TI - Induced Autolysis of Aspergillus oryzae (A. niger group): IV. Carbohydrates. AB - The examination of substances formed during induced autolysis by Aspergillus niger was continued in this work, which dealt in particular with carbohydrates. The autolysate contained a large amount of d-glucose (14 to 20% dry wt) and traces of glycolic aldehyde, dihydroxyacetone, ribose, xylose, and fructose. It also contained glycopeptides (about 10% dry wt), which were split from the cell wall during autolysis and which differed from one another in their level of polymerization and their composition. They were constituted by glucose and mannose, glucose and galactose, or mannose, glucose, and galactose (mannose being the most abundant in this case), and amino acids (chiefly alanine, serine, glutamic acid, and aspartic acid). During autolysis, only a part of the cell wall was dissolved, since it retained its shape. Upon further chemical hydrolysis, it produced mostly glucose and glucosamine, and smaller amounts of mannose, galactose, and amino acids. Presumably, glucomannoproteins and glucogalactoproteins were present in the intact cell as a macromolecular complex, constituting, together with chitin, the major part of the cell wall of Aspergillus. PMID- 16349625 TI - Microbiological Transformation of Terpenes: II. Transformation of alpha-Pinene. AB - Several strains of fungi were tested for their ability to metabolize alpha-pinene in shake cultures. A strain of Aspergillus niger showing marked efficiency in this respect was selected for further studies. The optimal conditions for fermentation were established with respect to substrate concentration, time, and temperature. From the fermentation products three major metabolites of alpha pinene were isolated: a ketone, C(10)H(14)O, identified as d-verbenone; an alcohol, C(10)H(16)O, identified as d-cis-verbenol; and a crystalline diol, C(10)H(18)O(2), characterized as d-trans-sobrerol. PMID- 16349624 TI - Biosynthesis of Cephalosporin C: I. Factors Affecting the Fermentation. AB - A series of complex and synthetic media have been developed that are suitable for the production of cephalosporin C and cephalosporin N by a mutant strain of Cephalosporium (C.M.I. 49,137). dl-Methionine increased the yield of both antibiotics, with more effect on cephalosporin N. l-Cystine had a greater enhancing effect on formation of cephalosporin C than on formation of cephalosporin N in synthetic media; serine increased yields of cephalosporin C under certain conditions. Disaccharides or polysaccharides appeared to be the best source for carbohydrates. No evidence was found for precursor action such as is found in penicillin fermentations. The ability of resting cells to produce antibiotic was demonstrated. PMID- 16349626 TI - Bacteriology of Manganese Nodules: I. Bacterial Action on Manganese in Nodule Enrichments. AB - Bacteria, found in manganese nodules from the Atlantic Ocean, enhance the adsorption of Mn from sea water by crushed manganese nodules in the presence of peptone. When bacterial outgrowth from crushed manganese nodules was experimentally delayed, peptone did not enhance Mn adsorption by nodular substance, but hindered it in some cases. A mechanism to explain the role of bacteria in enhancing Mn adsorption by manganese nodules is presented. Oyster shells were shown to adsorb Mn in the absence of bacteria. Peptone did not enhance the rate of Mn adsorption. Adsorbed Mn was not visibly oxidized during experimental observation. These results suggest one way whereby nodule formation may be initiated in the oceans. Some bacteria in the nodules were found to release manganese from them in the presence of glucose and peptone. Bacteria may, therefore, play a role not only in nodule buildup but also in nodule breakdown. PMID- 16349627 TI - Microbiological Leaching of Metallic Sulfides. AB - The percentage of chalcopyrite leached in percolators by Thiobacillus ferrooxidans was dependent on the surface area of the ore but not on the amount. Typical examples of ore leaching, which demonstrate the role of the bacteria, are presented. In stationary fermentations, changes in KH(2)PO(4) concentration above or below 0.1% decreased copper leaching as did reduction in the MgSO(4).7H(2)O and increase in the (NH(4))(2)SO(4) concentration. Bacterial leaching of chalcopyrite was more effective than nonbiological leaching with ferric sulfate; ferric sulfate appeared to retard biological leaching, but this effect was likely caused by formation of an insoluble copper-iron complex. Ferrous sulfate and sodium chloride singly accentuated both bacterial and nonbiological leaching of chalcocite but jointly depressed bacterial action. Sodium chloride appeared to block bacterial iron oxidation without interfering with sulfide oxidation. Bacterial leaching of millerite, bornite, and chalcocite was greatest at pH 2.5. The economics of leaching a number of British Columbia ore bodies was discussed. PMID- 16349628 TI - Variations in Strains of Streptomyces griseus Isolated from a Degenerating Streptomycin-Producing Culture. AB - Streptomycin-resistant strains were isolated from a degenerated streptomycin producing culture of Streptomyces griseus. From 250 resistant strains, 3 low, 2 intermediate, and 2 high potency strains were selected; these were compared in their morphological, cultural, physiological, and streptomycin-producing properties. Though no definite correlation between streptomycin production and the other properties could be obtained, the following correlations were considered as distinct differences among the low, intermediate, and high potency strains. (i) When streptomycin-producing ability degenerates, more submerged spore formation or fragmentation of mycelium into shorter filaments appears to occur. (ii) On agar medium, low and intermediate potency strains often show finely wrinkled growth; high potency strains do not show such characteristics. (iii) High potency strains excrete a distinct yellow soluble pigment on synthetic agar medium and on glucose-yeast extract agar, but low and intermediate potency strains show little or no ability to form this soluble pigment. (iv) In low and intermediate potency strains, inositol and arginine did not stimulate streptomycin production as they did in high potency strains. Streptamine showed some stimulating effect in the high potency strains and, in contrast, a depressive effect in intermediate potency strains, though streptidine showed a distinctly stimulating effect in all groups of strains employed. PMID- 16349629 TI - Effect of Pasteurization on Survival of Certain Psychrophilic Bacteria. AB - The effect of pasteurization on survival in milk of four strains of psychrophilic pseudomonads and of one strain of Alcaligenes was determined. Cell numbers were very low immediately after pasteurization, but increased after storage. The survival of psychrophilic bacteria in pasteurized milk appeared to depend on the initial number of organisms, and detection depended partly on the length of time of storage at 3 to 5 C. PMID- 16349630 TI - Fungi Associated with Softening of Bisulfite-Brined Cherries. AB - Softening of sound, calcium bisulfite-brined cherries was induced fairly quickly by brining them with cherries rotted by Aspergillus niger, Cytospora leucostoma, and Penicillium expansum, but not with cherries rotted by a variety of other microorganisms, including Alternaria sp., Aspergillus oryzae, Aureobasidium pullulans, Botrytis cinerea, Cladosporium sp., Mucor racemosus, Rhizopus stolonifer, and Sclerotinia fructicola. Rapid softening was correlated with the presence of a bisulfite-stable polygalacturonase, as demonstrated by a cup-plate test. A survey of naturally rotted cherries suggests the involvement of a bark canker fungus, C. leucostoma, in softening of commercially brined cherries in the Pacific Northwest. PMID- 16349632 TI - Amino Acid Composition of the Protein from a Mushroom (Pleurotus sp.). AB - Approximate analyses of mushroom protein (Pleurotus sp.) revealed that it contains 2.78% protein and 0.14% nonprotein nitrogen on a fresh-weight basis. A total of 17 amino acids, including all the essential amino acids, were qualitatively identified. Quantitative estimation of essential amino acids showed that, except for methionine and phenylalanine, all are in fairly high concentration. From these studies, it was concluded that the supplementation of this mushroom with cereal diet would help to overcome lysine deficiency. PMID- 16349631 TI - Effect of Environmental and Physiological Conditions on the Phase of Adjustment of Pseudomonas fragi. AB - The duration of the phase of adjustment of Pseudomonas fragi was affected by the physiological age and growth temperature of the inoculum, as well as by the temperature at which the growth curve was determined. Cultures in the exponential phase of growth gave shorter lags than stationary-phase and resting-phase inocula. Inocula from the latter phase gave the longest lags. Inocula grown at the temperature at which the growth curve was determined usually gave the shortest lags: the greater the difference between the incubation temperature of the inoculum and the incubation temperature of the growth curve, the longer the lag. Inocula grown at temperatures below the incubation temperature of the culture tended to produce longer lags than inocula grown at temperatures above the incubation temperature. The combined effect of physiological age and incubation temperature of the inoculum was additive. The effect of the incubation temperature of the culture upon the duration of the lag depended upon the method used to determine this phase. Lags that were measured in physical time (i.e., Lockhart's lag) decreased as the incubation temperature of the culture was increased, within the temperatures used. But Monod's lag, which measures physiological time, did not decrease as the temperature of growth increased but rather appeared to vary around some constant value dependent upon the physiological condition of the culture. PMID- 16349633 TI - Microbial Degradation of Asphalt. AB - Organisms of the genera Pseudomonas, Chromobacterium, and Bacillus capable of degrading asphalt were isolated by enrichment cultures. The asphalt degradation by these organisms varied from 3 to 25% after incubation for 1 week. The effects of temperature, pH, and atmosphere of incubation on asphalt degradation were investigated and were shown to vary with different organisms on the same substrate. PMID- 16349634 TI - Production of Oxalic Acid by a Wood-Rotting Fungus. AB - The wood-rotting fungus Pleurotus ostreatus NRRL 2366 was grown successfully in submerged shaker cultures in which it produced oxalic acid from simple carbohydrates as efficiently as did Aspergillus niger. P. ostreatus also produced oxalic acid from mixtures of sawdust and CaCO(3), and from the solid residue from the acid hydrolysis of wood when the culture was supplemented with inorganic nutrients. A. niger produced oxalic acid from the liquid hydrolysate. PMID- 16349635 TI - Production of Oxalic Acid by a Strain of Agaricus campestris. AB - A strain of the common mushroom Agaricus campestris was grown on a mixture of composted sawdust and CaCO(3). On incubation for 47 days, the organism produced 20.5 g of oxalic acid per 100 g of initial dry compost solids. PMID- 16349636 TI - Sorbic Hydroxamic Acid, an Antifungal Agent Effective over a Wide pH Range. AB - Sorbic hydroxamic acid was prepared from sorbic acid by esterification and treatment with hydroxylamine (mp 133 to 135 C, pK(a) 8.8). Its ultraviolet spectrum in acid solution had a single absorption maximum at 262 mmu; in alkaline solution the maximal absorption shifted to 255 mmu and significant absorption appeared at 280 to 300 mmu. At concentrations of 0.1% (w/v), sorbic hydroxamic acid prevented the growth of Aspergillus niger, Penicillium notatum, Botrytis cinerea, Cladosporium herbarum, and a Rhizopus species in grape juice over the pH range 3.6 to 9.2, although sorbic acid was not effective at pH 5.7 and above. PMID- 16349637 TI - Accuracy of a Plant-Infection Technique for Counting Populations of Rhizobium trifolii. AB - A "plant-infection" technique for identification and estimation of populations of Rhizobium is described and compared with petri dish colony counts of the same bacterial populations. Provided that bacterial suspensions are agitated thoroughly, dilutions made at 4 C, and test plants grown on agar, the plant infection technique is an accurate method of estimating R. trifolii in pure culture or when added to soil. The results are discussed in relation to previous investigations of the subject. Tables are presented which, when applied to the distribution of positive (nodulated) test plants in either fivefold or tenfold dilution series, permit calculation of most probable numbers, and confidence limits are stated. PMID- 16349638 TI - Production of Pectolytic Enzymes by Rhizopus stolonifer Sporangiospores After "Lethal" Gamma Irradiation. AB - Irradiated sporangiospores of Rhizopus stolonifer excreted pectolytic enzymes, which hydrolyzed pectin and macerated potato tuber discs, into the suspending medium. Pectin glycosidase, but not pectin methylesterase, activity developed regardless of the amount of radiation the spores had received, unless the dose exceeded about 1 megarad. The ability to produce pectolytic enzymes was found to be more radiation-resistant than the potential for colony formation or the ability to germinate. Spores made incapable, through irradiation, of forming colonies continued to produce pectolytic enzymes after a 6-day period following irradiation treatment. PMID- 16349639 TI - Viability of Selected Microorganisms in Hydrocarbon Fuels. AB - A laboratory study of the viability of selected microorganisms in a hydrocarbon fuel medium was carried out on 19 species of microorganisms, representative of the types found as natural contaminants in aircraft fuels. More species remained viable when inoculated in pure cultures than when inoculated in mixed (composite) cultures. Of the 19 species selected, 10 were still viable after 3 months and 5 were viable after 4 months in the pure culture inoculants. In the complete composite culture inoculant, the bacterial species which were viable at the end of 4 months were the same as those found in the pure culture inoculant. No fungi remained viable in the complete composite cultures after a 3-week period. The microorganisms which remain viable in a hydrocarbon fuel medium are considered indicative of a satisfactory inoculum to be used as a test culture in laboratory analysis of mechanical control techniques. PMID- 16349640 TI - Growth of Fungi in Sea Water Medium. AB - The effect of sea water on protein synthesis and growth of some species of Fungi Imperfecti was investigated. The dry weight of mycelia was greater from sea water medium than from distilled water medium in most instances; however, in a few there was a marked reduction in growth. The beneficial effect could not be ascribed to sodium choride or phosphate ion but may be due to the magnesium ion in sea water. This is indicated by the increase in mycelial yield and protein synthesis, even above that obtained with sea water, when magnesium ion was added to the medium. The reduction in yield and protein synthesis in some instances may be attributed to metal interactions; this possibility is being investigated further. PMID- 16349641 TI - Dialysis Fermentor Systems for Concentrated Culture of Microorganisms. AB - Cell-mass production of Serratia marcescens was studied in dialysis systems in which growth was managed in a fermentor remote from, but connected by conduits and pumps with, a nutrient reservoir. Dialysis was accomplished with membrane tubing in either the fermentor or the reservoir, or best with membrane sheet in a plate-and-frame dialyzer that was remote from but connected with both vessels. Growth trials with these systems demonstrated their ability to produce virtually unlimited population density in a liquid culture, viable counts in excess of 10 cells/ml and partial cell volume of 50% being attained. The system used for growth also may be used, after the growth cycle, to concentrate cells still further by osmotic dehydration with a hydrophilic colloid. The dialyzer-dialysis system that was evolved permits independent control of the component operations and is believed to be adaptable to any desired scale of size. PMID- 16349642 TI - Gnotobiotic Silage. AB - Selected strains of lactic acid bacteria isolated from grass silage were found to flourish when inoculated into irradiation-sterilized forage under gnotobiotic conditions. The acid content and pH of these silages resembled naturally fermented silage. Inoculation of gnotobiotic silage with Clostridium sporogenes and C. tyrobutyricum failed to cause any noticeable deterioration of silage quality. PMID- 16349643 TI - Violet-Pigmented Pseudomonads With Antifungal Activity From the Rhizosphere of Beans. AB - Bean rhizosphere bacteria antagonistic to four root-infecting fungi and an antibiotic produced by these bacteria were studied. The bacteria were violet pigmented gram-negative rods, probably belonging to the genus Pseudomonas. The antibiotic, which was localized largely in the bacterial cell mass, was easily extracted with acetone. It was selectively active against a wide variety of plant pathogenic and saprophytic fungi tested in vitro but was relatively inactive against bacteria. The compound, partially purified by chromatography, was soluble in all organic solvents tried, but nearly insoluble in water. It demonstrated no characteristic ultraviolet- or visible-absorption spectrum and was chemically unidentified. The antagonistic bacteria or crude antibiotic applied to buried buckwheat segments suppressed the colonization of this substrate by Rhizoctonia spp. The data suggested that the bacteria or the antibiotic may play a role in the suppression of root-infecting fungi in soil. PMID- 16349644 TI - Degradation of Wood Preservatives by Fungi. AB - Wood-inhabiting fungi, not necessarily responsible for major decay, are shown to be capable of degrading a toxic compound into a less potent form, thus rendering it less effective in protecting wood from decay by less-tolerant basidiomycetous wood-destroyers. Sweetgum or pine sapwood blocks treated with preservatives (ammoniacal copper arsenate, fluor-chrome-arsenate-dinitrophenol, a creosote or pentachlorophenol) were exposed progressively to two different wood-inhabiting fungi with sterilization between the first and second exposure. The fungus in the first exposure was usually an Ascomycete or a Fungi Imperfecti-Chaetomium globosum, Phoma, Orbicula, Graphium, Pestalozzia, or Trichoderma species, isolated from wood below the ground. In one experiment, the fungus in the first exposure was a basidiomycete, Lenzites trabea or Polyporus versicolor. The second fungus, a prominent Basidiomycete-Coniophora puteana, Lentinus lepideus, or Lenzites trabea-was the bioassay fungus, since its purpose was to show whether the first fungus had degraded the preservative. Generally, the treated block, except where exposed to another fungus, remained virtually untouched by the bioassay fungus. Clearly, therefore, the first fungus had rendered the preservative ineffective but without appreciably decaying the wood itself Chemical analyses of treated blocks indicated that in the first exposure the fungi had substantially depleted sodium arsenate and pentachlorophenol. PMID- 16349645 TI - Accuracy of a Plant-Infection Technique for Counting Populations of Rhizobium trifolii. AB - [This corrects the article on p. 379 in vol. 11.]. PMID- 16349646 TI - Selection of a Microbiological Corrosion System for Studying Effects on Structural Aluminum Alloys. AB - Two laboratory methods, a metal-strip test and a tank test, were evaluated as microbiological corrosion systems for producing corroded test specimens on a structural aluminum alloy. The results show that corrosion of the test alloy occurred best in the metal-strip test in a deionized water-fuel medium inoculated with a mixture of microorganisms under aerated conditions. The metal-strip test was more successful for producing large numbers of corroded test specimens and proved more economical than the tank-type test, since less structural material is needed to obtain a specimen with sufficient corrosion areas, and since the corrosion can more easily be restricted by maskants to certain areas for specific test purposes. PMID- 16349647 TI - Gelatinase Activity by Marine Fungi. AB - Gelatinase activity was detectable in the culture filtrates of 13 of 14 species of marine fungi tested. The fungi were grown under conditions of submerged culture in a medium consisting of corn meal, soybean meal, and CaCO(3). The degree of enzymatic activity did not necessarily correlate with the amino nitrogen content of the filtrates. Eight of the filtrates contained measurable quantities of glutamic acid and alanine. Highest levels of gelatinase were obtained in the culture filtrates of Halosphaeria mediosetigera. Increasing the corn meal concentration of the medium resulted in even greater production of gelatinase by H. mediosetigera, whereas amino acid yields were enhanced by higher concentrations of soybean meal. Supplementation of the high corn meal medium with either a mineral mixture or distillers' solubles brought about a faster rate of production of gelatinase and, in the case of the distillers' solubles, a significantly higher level of enzyme, as well. PMID- 16349648 TI - beta-Glucosidase Activity in Phytopathogenic Bacteria. AB - Most of 58 isolates of phytopathogenic and related bacteria comprising 24 species in the genera Agrobacterium, Erwinia, Corynebacterium, Pseudomonas, and Xanthomonas exhibited beta-glucosidase activity, especially the gall-nonforming pathogenic pseudomonads and soft rot organisms. The gall-forming pseudomonads and P. fluorescens exhibited no beta-glucosidase activity, with the exception of one isolate of P. savastanoi which showed slight activity on an inorganic nitrogen arbutin medium. The best medium for demonstrating beta-glucosidase activity contained peptone as the nitrogen source and arbutin. beta-Glucosidase activity in this medium was indicated by either acid production or browning. P. syringae, in contrast to other bacteria tested, produced most beta-glucosidase in a medium containing large amounts of glucose. Chromatographic analyses confirmed that splitting of the glucoside occurred at the glucosidic linkage. Reaction of sonically treated bacterial cells with indican or p-nitrophenyl-beta-D-glucoside proved a rapid method for assaying relative amounts of beta-glucosidase among bacterial species. Harda's paper-strip method of detecting beta-glucosidase also was useful in revealing the distribution and relative amounts of beta-glucosidase in most bacteria, but did not indicate the relatively greater amount of beta glucosidase in P. syringae. PMID- 16349650 TI - Lipid Alterations During the Fermentation of Dill Pickles. AB - Analyses of various lipid fractions of sections of cucumbers and of good and bloated dill pickles showed that marked changes occur in all lipid fractions during fermentation. The most striking difference noted was the decrease in the phospholipid fraction. A nearly fourfold increase in free fatty acid, as well as a marked increase in the neutral fat fatty acids and unsaponifiables, occurred. Gas chromatographic analyses of the methyl esters of fatty acids from the various lipid fractions yielded further interesting data. From the analyses, 41 esters were identified; however, 16 of the esters accounted for at least 95% of the acids. Among the marked changes were the increases in linoleic and linolenic acids in good pickles, in contrast to the increase in oleic acid in the bloated pickles. The presence of tridecenoic acid in cucumbers, and its absence in pickles; and the absence of caproic, caprylic, and capric acids in cucumbers, and its presence in pickles, were interesting. The data demonstrated that the lipid alterations that occur during fermentation of cucumbers are analogous to those previously reported for the sauerkraut fermentation. PMID- 16349649 TI - Antibiotics Against Plant Disease: VIII. Screening for Nonpolyenic Antifungal Antibiotics Produced by Streptomycetes. AB - In a survey of Streptomyces species, methods were designed and followed that would specifically select strains capable of producing heat-stable, nonpolyenic, antifungal antibiotics. Of 500 strains grown in shaken flasks, 240 of the culture liquors contained active factors as demonstrated by paper-disc assay against Mucor ramannianus. Culture filtrates and mycelial extracts of the active strains were examined by ultraviolet spectrophotometry; 166 were nonpolyenic as determined by absorption spectra. Heat-stability tests of the nonpolyenic antibiotics over a broad pH range revealed that 15 were stable under all test conditions, 70 moderately stable, and 81 unstable. Culture liquors containing stable, nonpolyenic antifungal agents were chromatographed with eight solvent systems in an attempt to identify the antibiotics. The producing cultures were studied by cross-antagonism tests to discover similarities with producers of known antibacterial antibiotics. Two of the antibiotics produced by promising strains were identified as cycloheximide and musarin. Six antibiotics, presumably new, were detected. PMID- 16349651 TI - Pure Culture Fermentation of Brined Cucumbers. AB - The relative abilities of Pediococcus cerevisiae, Lactobacillus plantarum, L. brevis, and several other species of lactic acid bacteria to grow and produce acid in brined cucumbers were evaluated in pure culture fermentations. Such fermentations were made possibly by the use of two techniques, gamma radiation (0.83 to 1.00 Mrad) and hot-water blanching (66 to 80 C for 5 min), designed first to rid the cucumbers of naturally occurring, interfering, and competitive microbial groups prior to brining, followed by inoculation with the desired lactic acid bacteria. Of the nine species tested, strains of the three common to cucumber fermentations, P. cerevisiae, L. plantarum, and L. brevis, grew to the highest populations, and produced the highest levels of brine acidity and the lowest pH values in fermentations at 5.4 to 5.6% NaCl by weight; also, their sequence of active development in fermentations, with the use of a three-species mixture for inoculation, was in the species order just named. This sequence of occurrence was similar to that estimated by others for natural fermentations. The rates of growth and acid production in fermentations with a mixture of P. cerevisiae, L. plantarum, and L. brevis increased as the incubation temperature was increased from 21 to 27 to 32 C; however, the maximal populations and acidities attained were essentially the same for fermentations at each temperature. Further, these same three species were found to be the most salt tolerant of those tested; their upper limit for appreciable growth and measurable acid production was about 8% salt, whereas thermophilic species such as L. thermophilus, L. lactis, L. helveticus, L. fermenti, and L. delbrueckii exhibited a much lower salt tolerance, ranging from about 2.5 to 4.0%. However, certain strains of L. delbrueckii grew very rapidly in cucumbers brined at 2.5 to 3.0% salt, and produced sufficient acid in about 30 hr at 48 C to reduce the brine pH from above 7.0 to below 4.0. An inexpensive, pure culture fermentor which was suitable for gamma radiation, resistant to salt and acid, and which permitted repeated aseptic sampling of the fermenting brine, is illustrated and the specifications are given. PMID- 16349652 TI - Microbial Degradation of the Sulfonate of Dodecyl Benzene Sulfonate. AB - It has been observed that the sulfonate portion of alkyl benzene sulfonate (ABS) will undergo microbial attack in certain pure and mixed cultures if an energy source, such as glucose, is available. The evidence for this is provided by the stoichiometric relationship between the reduction of ABS concentration and the appearance of inorganic sulfur compounds. PMID- 16349653 TI - Kinetics of Death of Bacterial Spores at Elevated Temperatures. AB - [This corrects the article on p. 452 in vol. 12.]. PMID- 16349654 TI - Fungistatic Performance of 10,10'-Oxybisphenoxarsine in Exterior Latex and Asphalt Coatings. AB - 10, 10'-Oxybisphenoxarsine has been found to be outstanding in its activity against bacteria and fungi. Parallel tests with known fungistats for comparison (2,3,4,6-tetrachlorophenol and phenylmercuric acetate) have demonstrated its superior activity and persistence in an exterior acrylate paint film and in an asphalt coating. In view of its superior antimicrobial activity and its persistence, it can be used in applications in which there is no danger of its ingestion. Further field trials are in progress to evaluate 10, 10'oxybisphenoxarsine in paint, asphalt, wood, and in marine pilings. PMID- 16349655 TI - NEW ANALAB SERIES OSCILLOSCOPES FEATURE DIGITAL SWEEP DELAY AND READOUT. PMID- 16349656 TI - PEPTIDE ANALYZER. PMID- 16349658 TI - In Vitro and In Vivo Laboratory Evaluation of Cephaloglycin and Cephaloridine. AB - [This corrects the article on p. 249 in vol. 13.]. PMID- 16349657 TI - Cultivation of Pasteurella haemolytica in a Casein Hydrolysate Medium. AB - The growth of Pasteurella haemolytica strain H44L was studied under aerobic conditions in a medium of acid-hydrolyzed casein, supplementary cysteine, inorganic salts, vitamins, and a carbon source. The concentration of casein hydrolysate necessary for optimal growth was 1.5 or 2.0%, depending upon the carbon source employed. Essential vitamins were calcium pantothenate, nicotinamide, and thiamine. Concentrations as low as 0.01 mug/ml of thiamine monophosphate or thiamine pyrophosphate supported maximal growth, but thiamine hydrochloride or thiamine nitrate were active only at the unusually high levels of 10 to 20 mug/ml. The best carbon sources were d-galactose or sucrose. Maximal growth resulted from an inoculum containing fewer than 10 cells per milliliter of medium. Cellular yields averaged 6 x 10 to 7 x 10 cells per milliliter for the test organism and five other strains of P. haemolytica isolated from cases of bovine respiratory diseases. PMID- 16349659 TI - Pouch Method for the Isolation and Enumeration of Clostridia. AB - [This corrects the article on p. 281 in vol. 13.]. PMID- 16349660 TI - Role of Leuconostoc mesenteroides in Leavening the Batter of Idli, a Fermented Food of India. AB - [This corrects the article on p. 227 in vol. 13.]. PMID- 16349661 TI - Detection of Minimal Quantities of Foot-and-Mouth Disease Virus with Bovine Kidney Tissue Cultures. AB - Minute quantities of foot-and-mouth disease virus (FMDV) were suspended in 500-, 1,000- and 2,000-ml volumes of virus growth medium and inoculated into primary bovine kidney cell cultures grown as monolayers in 5-liter Povitsky bottles. After 48 hr of incubation, the low concentrations of FMDV, containing from 10 to 10 plaque-forming units (PFU) per milliliter of inoculum, multiplied to approximately 10 PFU per milliliter of fluid. The presence of extracellular virus after multiplication was verified by assaying the fluid in 4-oz prescription bottle cultures. Minimal quantities of FMDV were able to infect the cells and replicate despite the large volume of the suspending fluid. This technique may be applicable for the detection of trace amounts of FMDV in clinical samples and vaccines. Skin samples of unknown infectivity in limited tests gave identical positive or negative results in both steers and Povitsky cultures, but were consistently negative when tested in suckling mice and in prescription-bottle tissue cultures. PMID- 16349662 TI - Steroid mechanisms affecting yeast permeability and viability. AB - A synthetic aza-cholestane amine was found to be rapidly lethal to Saccharomyces cerevisiae. Lethal action was prevented by pretreating cells with 10m uranyl nitrate. Sublethal concentrations of the steroid prevented the uptake of glucose and alanine. This demonstrated one means by which the steroid affected cell permeability, limiting the growth of the organism. Decreases in viability were related to increases in 260 and 280 mmu absorbing materials. Uranyl protection of viability was found to be related to cellular retention of the ultraviolet absorbing materials. The steroid, dequadin acetate, and cetyl pyridinium chloride were comparable in lethal action and in causing the leakage of cytoplasmic constituents. Loss of viability and cytoplasmic excretion were detected within minutes after cells were exposed to the steroid. The rate or degree of metabolism of glucose by resting cells did not appear to influence steroid action or the sensitivity of the cells to the steroid. PMID- 16349663 TI - Biochemical Alterations of Dermatophytes during Growth. AB - Alterations in the biochemical constituents of mycelia were studied during the growth, development, and starvation of Microsporum quinckeanum. On the basis of dry weight, growth of this dermatophyte could be divided into four phases: lag, log, stationary, and death. The percentage of total nitrogen, inorganic phosphorus, ribonucleic acid (RNA), and protein increased rapidly during the lag phase. The percentage of protein remained constant after the initial increase; however, inorganic phosphate and RNA decreased in older mycelia. Acid-soluble materials in the cells increased in concentration as the organism aged. Chitin was present in the spores at a much higher concentration than in the mycelia. The percentage of this compound decreased rapidly until the end of the lag phase. An increase and subsequent decrease in per cent chitin occurred during the log phase. Inorganic phosphorus in the mycelia increased from the value in the spore stage to a maximum in the early log phase, and then decreased rapidly during the remainder of the growth cycle. Compounds involved in protein synthesis increased rapidly during the lag phase of growth. Changes in chemical composition of the mold during starvation indicate that carbohydrate does not form the principal endogenous reserve of M. quinckeanum, whereas lipids may represent the primary reserve material. PMID- 16349664 TI - Modified gradient plate for use in the virus plaque technique. PMID- 16349665 TI - Production of the Enzyme Naringinase by Aspergillus niger. AB - [This corrects the article on p. 842 in vol. 13.]. PMID- 16349666 TI - Durhamycin, a Pentaene Antifungal Antibiotic from Streptomyces durhamensis sp. n. AB - Durhamycin, derived from a previously undescribed soil isolate, is an apparently new antibiotic active against many of the fungi pathogenic for man and animals. Cultural and morphological characteristics and biochemical reactions of Streptomyces durhamensis are detailed. Methods of extraction of the antibiotic, physicochemical properties, antimicrobial spectrum, and results of mouse toxicity and protection tests are given. PMID- 16349667 TI - Production of guanosine-5'-monophosphate and inosine-5'-monophosphate by fermentation. AB - A biotin-requiring coryneform bacterium which produces glutamic acid was mutated to adenine dependency. The adenine-requiring strain, which excreted insoine-5' monophosphate (IMP), was further mutated to xanthine dependency. As expected, IMP was also excreted by this mutant. The mutant strain was reverted to xanthine independence in an attempt to obtain a culture with an altered IMP dehydrogenase which would be less sensitive to feedback inhibition by guanosine-5' monophosphate (GMP). A revertant was obtained which produced GMP and IMP, each at 0.5 g per liter. The reversion to xanthine independence had resulted in a concomitant requirement for isoleucine, leucine, and valine. Further mutation to increased nutritional requirements led to culture MB-1802, which accumulated 1 g per liter each of GMP and IMP. Both nucleotides were isolated in pure form. The concentrations of GMP and IMP produced by MB-1802 were four times that of cytidylate, uridylate, or adenylate, indicating that the mechanism of GMP and IMP production was direct and not via ribonucleic acid breakdown. PMID- 16349668 TI - Identification of fluorescent-antibody labeled group a streptococci by fluorometry. AB - [This corrects the article on p. 386 in vol. 14.]. PMID- 16349669 TI - Conversion of C-labeled substrates to volatile Fatty acids by the rumen microbiota. AB - The fermentation of uniformly labeled glucose-C, glucose-1-C, -2-C, and -6-C, xylose-1-C, cellulose-1-C, -2-C, and -6-C, and lactate-2-C by rumen fluids from cows fed all-hay, hay and concentrate (50:50), and all-concentrate diets was investigated. The results obtained suggested that the Embden-Meyerhof glycolytic pathway is the major pathway of hexose utilization, that the major pathway of xylose fermentation involves hexose synthesis, and that the contributions of the nonrandomizing (acrylate) pathway of propionate formation during glucose, xylose, and cellulose fermentations are 4.5, 8.0, and 10.5%, and 24.6, 25.8, and 17.2%, respectively, by rumen fluids from the cows fed all-hay and all-concentrate rations. PMID- 16349670 TI - Western equine encephalomyelitis vaccine produced in chick embryo cell cultures. AB - A method was developed for production of a freeze-dried Western equine encephalomyelitis vaccine from virus propagated in chick embryo cell culture monolayers maintained with a serum-free medium. A sufficient concentration of virus accumulated in the cell culture fluids prior to the occurrence of viral cytopathology to permit the production of a vaccine relatively free from serum and cellular proteins. Inoculation with two mouse ld(50) doses of virus per 100 tissue culture cells was found to yield reproducible high virus titers at a convenient harvest time. These harvests were inactivated at 22 C by 0.05% formalin within 48 hr. Potency test results, as measured by the protection of immunized guinea pigs against an intracerebral virus challenge, indicated that the vaccine produced from the virus propagated in cell culture was equal in potency to a lot of whole chick embryo vaccine used to immunize laboratory and field workers subject to a high risk of infection. PMID- 16349671 TI - Solubilization of Acid-swollen cellulose by an enzyme system from a species of alternaria. AB - An unknown species of Alternaria, when grown on a medium containing carboxymethylcellulose as a carbon source produced a mixture of extracellular enzymes which solubilized acid-swollen cellulose. The product of the hydrolysis was a 1:2 molar mixture of cellobiose and glucose. The organism apparently produced no cellobiase. It is suggested that the mixture of cellulolytic enzymes contains at least two different enzymes which degrade cellulose in an endwise manner. PMID- 16349672 TI - Effect of Aeration on Growth and Aflatoxin Production by Aspergillus flavus in Submerged Culture. AB - Aspergillus flavus ATCC 15517 produced up to 212 mg per liter of total aflatoxin in submerged culture in aerated (3,000, 6,000, 9,000, and 12,000 ml/min) and agitated medium in 14-liter fermentors with 10 liters of medium consisting of 2% yeast extract and 10% sucrose. Aflatoxin production increased with time. A maximum of 212 mg/liter was produced at 9,000 ml/min aeration, whereas the yield decreased substantially at the lower aeration rates. Two other strains of A. flavus synthesized aflatoxin in smaller quantities. PMID- 16349673 TI - Amino Acid and vitamin requirements of several bacteroides strains. AB - Nutritional studies were performed on nine Bacteroides strains, by use of the methodology and media of anaerobic rumen microbiology. Ristella perfoetens CCI required l-arginine hydrochloride, l-tryptophan, l-leucine, l-histidine hydrochloride, l-cysteine hydrochloride, dl-valine, dl-tyrosine, and the vitamin calcium-d-pantothenate, since scant turbidity developed in media without these nutrients. R. perfoetens was stimulated by glycine, dl-lysine hydrochloride, dl isoleucine, l-proline, l-glutamic acid, dl-alanine, dl-phenylalanine, dl methionine, and the vitamins nicotinamide and p-aminobenzoic acid, since maximal turbidity developed more slowly in media without these nutrients than in complete medium. Medium A-23, which was devised for R. perfoetens, contained salts, 0.0002% nicotinamide and calcium d-pantothenate, 0.00001% p-aminobenzoic acid, 0.044% l-tryptophan, 0.09% l-glutamic acid, and 0.1% of the other 13 amino acids listed above. Zuberella clostridiformis and seven strains of R. pseudoinsolita did not require vitamins, and showed no absolute requirement for any one amino acid. Various strains produced maximal turbidity more slowly in media deficient in l-proline, glycine, l-glutamic acid, dl-serine, l-histidine hydrochloride, dl alanine, or l-cysteine hydrochloride, than in complete medium. These eight strains grew optimally in medium A-23 plus 0.1% dl-serine but without vitamins. PMID- 16349674 TI - Pure culture fermentation of green olives. AB - The method previously developed by us for the pure-culture fermentation of brined cucumbers and other vegetables has been applied successfully to Manzanillo variety olives. Field-run grade fruit was processed first by conventional procedures to remove most of the bitterness. Then the relative abilities of Lactobacillus plantarum, L. brevis, Pediococcus cerevisiae, and Leuconostoc mesenteroides to become established and produce acid in both heat-shocked (74 C for 3 min) and unheated olives, brined at 4.7 to 5.9% NaCl (w/v basis), were evaluated. The heat-shock treatment not only proved effective in ridding the fruit of naturally occurring, interfering, and competitive microbial groups prior to brining and inoculation, but also made the olives highly fermentable with respect to growth and acid production by the introduced culture, particularly L. plantarum. Of the four species used as inocula, L. plantarum was by far the most vigorous in fermentation ability. It consistently produced the highest levels of brine acidity (1.0 to 1.2% calculated as lactic acid) and the lowest pH values (3.8 to 3.9) during the fermentation of heat-shocked olives. Also, L. plantarum completely dominated fermentations when used in two-species (with P. cerevisiae) and three-species (with P. cerevisiae and L. brevis) combinations as inocula. In contrast, when L. plantarum was inoculated into the brines of unheated olives it failed to become properly established; the same was true for the other species tested, but even to a more pronounced degree. L. brevis was the only species used that failed to develop in brines of both heat-shocked and unheated olives. Modification of the curing brine by the addition of lactic acid at the outset, either with or without dextrose, led to a much earlier onset of fermentation with accompanying acid development, as compared to treatments with dextrose alone or nonadditive controls. Reasons for the marked improvement of the fermentability of Manzanillo olives receiving the prebrining heat-shock treatment are discussed. PMID- 16349675 TI - RACK permitting efficient handling of tissue cultures. PMID- 16349676 TI - Suppression of Leuconostoc mesenteroides During Isolation of Lactobacilli. PMID- 16349677 TI - Dispensing Apparatus for Controlling E(h), pH, and Volume of Anaerobic Bacterial and Protozoal Culture Media. PMID- 16349678 TI - Acylation of 6-Aminopenicillanic Acid by Penicillium chrysogenum. PMID- 16349679 TI - Effect of various inhibitors of protein and deoxyribonucleic Acid synthesis on the growth of mycoplasmas. PMID- 16349680 TI - Preparation of Type-specific Antisera to Reoviruses. PMID- 16349681 TI - Instrumented quantitation of live bacteria in the presence of dead cells. PMID- 16349682 TI - Potency of adenovirus vaccines: comparison of unconcentrated, concentrated, and soluble antigen preparations. AB - Five adenoviruses were effectively concentrated and partially purified by methanol precipitation. The soluble antigens from one adenovirus concentrate were obtained by adsorption to and elution from calcium phosphate. Formaldehyde inactivated vaccines prepared from the virus concentrates were more antigenic in guinea pigs than a National Institutes of Health reference vaccine or one prepared from unconcentrated adenovirus. The soluble antigens also proved to be antigenic. The data indicate that adenovirus vaccines of superior potency can be prepared from concentrated virus preparations and that the extracted soluble antigens are immunogenic. PMID- 16349683 TI - Statistical evaluation of electronic and plate counts for estimating bacterial populations. AB - Bacteria from cultures of Staphylococcus aureus, Pseudomonas geniculata, and Rhodotorula glutenis were counted by use of an electronic counter and by plate counts from broth cultures at selected periods from 0 to 24 hr. Variations in the two methods were noted, and the results were compared after calculating correlations, coefficients of variation, and nested analyses of variance. It was not possible to determine the absolute accuracy of the two methods; however, the precision of the results obtained with an electronic counter was better than that obtained with plate counting. It appears that electronic counting, because of ease and convenience, provides better comparative counts at a particular stage of growth, especially during the early stages of the growth period, whereas plate counting yields the best results in determining growth curves. PMID- 16349684 TI - Action of chloramphenicol and its isomers on secondary biosynthetic processes of bacillus. AB - Chloramphenicol or its isomers, if supplied within 4 hr after the addition of Mn, prevented sporulation of Bacillus megaterium at concentrations that partially inhibited protein synthesis but which were neither bacteriostatic nor bactericidal. Likewise, sub-bactericidal quantities of the compounds, if supplied within 2 hr after the addition of Mn, suppressed formation of bacitracin by B. licheniformis. In contrast to previous reports that chloramphenicol is less active than its isomers against sporulation and peptide formation, the results of the present study indicated that the order and extent of these activities of the compounds is similar to that of their ability to prevent growth and to suppress protein synthesis; i.e., d(-)-threo > l(+)-erythro > d(-)-erythro. PMID- 16349685 TI - Glucose disappearance in biological treatment systems. AB - Laboratory scale anaerobic and aerobic treatment units were conditioned with a daily slug-feed of glucose. After a period of acclimation and stabilization, glucose disappearance was monitored continuously after the slug feed. A continuous sampling apparatus is described. Mathematical analysis of the data indicate zero-order reactions for both biological treatment systems. PMID- 16349686 TI - Steroid interference with antifungal activity of polyene antibiotics. AB - Wide differences exist among the polyene antibiotics, nystatin, rimocidin, filipin, pimaricin, and amphotericin B, with reference to steroid interference with their antifungal activities against Candida albicans. Of the numerous steroids tested, ergosterol was the only one which effectively antagonized the antifungal activity of all five polyene antibiotics. The antifungal activities of nystatin and amphotericin B were the least subject to vitiation by the addition of steroids other than ergosterol, and those of filipin, rimocidin, and pimaricin were the most sensitive to interference. Attempts to delineate the structural requirements of steroids possessing polyene-neutralizing activity in growing cultures of C. albicans are discussed. The ultraviolet absorbance of certain antibiotic steroid combinations was also studied. PMID- 16349687 TI - Formation of the blue pigment indigoidine by phytopathogenic erwinia. AB - Most cultures belonging to the "Erwinia chrysanthemi group" of soft-rot bacteria form traces of a blue, extracellular, water-insoluble pigment. This pigment is generally not found in cultures of the other members of the genus Erwinia. The blue substance has been isolated and purified from three members of the E. chrysanthemi group; it has been identified as indigoidine, 5,5'-diamino-4,4' dihydroxy-3,3'-diazadiphenoquinone-(2,2'). PMID- 16349688 TI - Inhibition of Swarming of Proteus by Sodium Tetradecyl Sulfate, beta-Phenethyl Alcohol, and p-Nitrophenylglycerine. AB - The effects of sodium tetradecyl sulfate (STS), beta-phenethyl alcohol (PEA), and p-nitrophenylglycerol (PNPG) on motility, swarming, flagellation, and growth of Proteus were examined. Growth-inhibitory concentrations (GIC) and swarming inhibitory concentrations (SIC) were determined. A characterization of the swarming-inhibitory efficacy of these compounds was based on their GIC/SIC ratio and their concentration inhibition curves. Using the homologous series of sodium alkyl sulfates as a standard reference, we showed that PNPG was more effective than STS, which was the most effective of the homologous series. PEA was less effective than sodium decyl sulfate but more effective than sodium octyl sulfate. Motility tests in liquid medium and electron microscope investigations indicated that the modes of action of the three compounds, all of which effectively inhibit the swarming of Proteus, are different. Whereas STS and PEA inhibit swarming by inhibition of motility, PNPG seems to act on the swarming mechanism sensu strictori, without impairment of motility. STS immobilizes by inhibition of flagellum formation or by some lytic action on the flagella already synthesized. PEA acts by impairing flagellar function, but leaves the flagella morphologically intact. PMID- 16349689 TI - Prevention of the emergence of drug resistance in bacteria by acridines, phenothiazines, and dibenzocycloheptenes. AB - It has been found that, like Atabrine, the phenothiazine tranquilizers promazine, chlorpromazine, promethazine, levopromethazine, and Stelazine; the antidepressants (dibenz-azepine and dibenzocycloheptene derivatives) Tofranil, Pertofrane, cyclobenzaprine, Elavil, protriptyline, and 3 chlorodibenzocycloheptene; and the acridine derivatives acridine orange, acriflavin, SKF no. 16214-A2, SKF no. 13231-A2, SKF no. 9200, and SKF no. 9836 are all to a greater or lesser extent than Atabrine, effective in preventing the emergence of resistant strains of Staphylococcus aureus and Escherichia coli in the presence of streptomycin, sulfathiazole, or chloramphenicol. The antimalarials chloroquine and hydroxychloroquine were also studied and found to be ineffective. The medical significance of these findings is discussed, as well as the effect of the structural variations of these compounds on their relative activities. PMID- 16349690 TI - Comparison of acetate differential agar with sellers citrate-mannitol-agar. AB - A total of 245 isolates of Escherichia coli (paracolons) were employed in comparing the use of acetate differential agar (ADA) with the simple test described by Sellers to eliminate these organisms from consideration as enteric pathogens. At 18 hr after the media were inoculated, 83% of the isolates produced a positive citrate reaction on Sellers citrate-mannitol-agar (SCM) and 7% gave a positive reaction on ADA. At 24 hr, 92% of the cultures were positive on SCM versus 42% on ADA. At 48 hr, 244 (99.6%) of the isolates were positive on SCM, whereas a 72-hr incubation period was required for equal results with ADA. One isolate failed to grow on either medium. PMID- 16349691 TI - Effect of air ions on submicron t1 bacteriophage aerosols. AB - The effect of a high concentration of ionized air molecules on sampling T1 phage aerosols of submicron particle size was evaluated by comparing the phage recoveries of all-glass impingers (AGI-4) and type 6 filter papers. Sampler recoveries of all ionized aerosols were less than the recoveries of nonionized control aerosols. These reductions in recovery were greater with positive ions than with negative ions or ions of mixed polarity. The AGI-4 allowed considerable slippage, which was not affected by the air ions. Type 6 filter paper recoveries were less than AGI-4 recoveries. The air ions did not appear to affect the aerosol particle size as determined by an electron microscope. PMID- 16349692 TI - Simple cultural test for relative cellulolytic activity of fungi. AB - A simple method is described for determining the relative cellulolytic activity of fungi. Opaque columns of an agar medium containing a partially crystalline cellulose preparation were inoculated with the fungi. Depth of the clear zone that developed beneath the growing cultures provided a visual measure of cellulolytic activity on a continuous, cumulative basis. Depth of clearing (DC) was determined for 25 species of fungi differing widely in cellulolytic activity, and compared by correlation analysis with results of three other methods for measuring cellulolytic activity. Relatively high coefficients of correlation (greater than 0.6) were obtained between DC and weight loss of cotton sliver, loss in tensile strength of cotton duck, and carboxymethyl cellulase activity in culture filtrates. In comparison with conventional assay procedures, the clearing method offered several advantages: (i) results were at least as well correlated with the capacity to utilize native cellulose as a substrate; (ii) the method measured activity of growing cultures rather than culture filtrates, thus involving less risk of losses due to product inhibition, binding, or denaturation of enzymes; (iii) repeated measurements were made on the same experimental set up, so that errors due to arbitrarily selected times of harvest were avoided conveniently; and (iv) the method required less working time and very simple equipment, making it convenient for large-scale screening tests. PMID- 16349693 TI - Adaptation of Pseudomonas fluorescens to Low Levels of Water Activity Produced by Different Solutes. AB - A culture of Pseudomonas fluorescens which grew at a minimal a(w) of 0.97 in a glucose-salts broth adjusted to that level with NaCl, KCl, or glucose, and at a(w) 0.9725 with Na(2)SO(4), was adapted to grow at lower a(w) levels produced by these solutes. Adaptation to growth at a low a(w) produced by means of one solute did not necessarily result in growth at that a(w) when another solute was employed to lower the a(w). The culture adapted to grow at a(w) 0.9650 produced by addition of glucose could grow equally well in broth adjusted to that a(w) with NaCl or KCl, but not Na(2)SO(4). NaCl and KCl permitted growth at the lowest a(w) level (0.9625), and the culture adapted in KCl did equally well in NaCl. PMID- 16349694 TI - Effect of Mg on the Activity of Vancomycin Against Escherichia coli. AB - Mg ions alleviate the inhibitory effect of vancomycin on Escherichia coli. This is not due to the formation of an antibacterial-inactive complex. It is suggested that vancomycin and Mg compete for a receptor site, or sites, on (or in) the bacterial cell. PMID- 16349695 TI - Effect of electrostatic charge on the contamination of plastic food containers by airborne bacterial spores. AB - Electrostatic charge of approximately -10 kv was produced by friction on polystyrene food container samples. This charge quickly decayed to a lower, more stable, level. Exposure of samples to positively charged red and negatively charged green fluorescent particles resulted in a particle-distribution pattern on the plastic surface. The dynamic attraction of fluorescent particles was illustrated by time-lapse photography. Similar distribution patterns of airborne bacterial spores were shown to develop. In controlled bacterial aerosol exposure tests, an increase in surface contamination of the plastic samples was found to be quantitatively related to an increase in negative electrostatic charge on the plastic. Static charge was found to accumulate on plastic food containers during their manufacture, and to remain indefinitely on many of the finished products. This charge was of the intensity and polarity to attract positively charged bacterial cells if such particles were present in the air. PMID- 16349696 TI - Survival of Clostridium perfringens During Baking and Holding of Turkey Stuffing. AB - Vegetative cells of three strains of Clostridium perfringens were used as inoculum for bread and onion stuffing for eight lightweight and eight heavyweight turkeys. When stuffed turkeys were refrigerated (5 +/- 1 C for 24 +/- 2 hr), a mean count of 580 vegetative cells of C. perfringens per gram of stuffing was reduced to undetectable levels (<6 per gram) in six of the eight. An inoculum of spores of the three strains used in a second series survived refrigerated holding with no change in numbers. During cooking of the stuffed turkeys in an oven at 94 C, numbers of vegetative cells fell steadily and numbers of spores remained constant or increased slightly (2 of 16 stuffings), until the temperature of the stuffing rose above that permitting growth. Viable C. perfringens cells were recovered from the stuffings at the end of cooking plus 1 hr for the group inoculated with the spore suspension. Storage of these stuffings resulted in marked reductions in numbers after 6 days at 5 +/- 1 C and in increases after 24 +/- 2 hr at 23 +/- 1 C. Cells of a strain which produces spores not considered heat-resistant survived in stuffing in birds cooked to doneness in ovens at 94, 163, and 232 C. In accepted methods of cooking stuffed turkeys, C. perfringens contaminants may survive and create a hazard if subsequent storage is in a temperature range which permits their multiplication. PMID- 16349697 TI - Changes in the Microflora of Vacuum-packaged, Irradiated Petrale Sole (Eopsetta jordani) Fillets Stored at 0.5 C. AB - The microfloral changes of irradiated petrale sole fillets during anaerobic (vacuum-packed in cans) refrigerated storage was determined by the identification of 1,260 microbial isolates to the generic level. The samples were irradiated at 0.0, 0.1, 0.2, 0.3, and 0.4 Mrad from a cobalt-60 gamma source, stored at 0.5 C, and examined periodically for spoilage and total microbial population and composition. The preirradiation flora consisted of Achromobacter, Micrococcus, Pseudomonas, coryneforms, Lactobacillus, and Flavobacterium. Immediately after irradiation, Micrococcus, Achromobacter, coryneforms, and Bacillus were predominant. After storage under vacuum, the spoilage flora of the nonirradiated petrale sole was predominantly Pseudomonas; the spoilage flora of the 0.1-Mrad irradiated samples consisted of Pseudomonas and Lactobacillus; and that of the 0.2-, 0.3-, and 0.4-Mrad samples was almost entirely Lactobacillus. PMID- 16349698 TI - Examination of Feces from Food Handlers for Salmonellae, Shigellae, Enteropathogenic Escherichia coli, and Clostridium perfringens. AB - Duplicate fecal specimens from food handlers were collected in Louisiana. One set of specimens was examined immediately for salmonellae and shigellae by the Central Laboratory of the Louisiana State Board of Health in New Orleans; the other set was shipped to the Food Microbiology Unit at the Robert A. Taft Sanitary Engineering Center in Cincinnati, Ohio, where it was examined for enteropathogenic Escherichia coli (EEC) and Clostridium perfringens. A total of 219 specimens were examined by both laboratories. None yielded salmonellae or shigellae; 171 (78.1%) yielded C. perfringens; 175 (79.9%) yielded E. coli; and 14 (6.4%) yielded EEC. The 14 isolates of EEC were distributed among eight serotypes; one specimen yielded two serotypes. Multiple isolations of C. perfringens strains (two to four) were made from 64 (37.4%) of the specimens, and a total of 244 strains were isolated and studied for identifying characteristics. Of the total, only 87 (35.5%) could be identified serologically by a battery of 67 antisera; only 4 (1.6%) possessed the characteristics of the English "food poisoning type." The hemolytic activity on agar containing horse, ox, or sheep blood showed that 140 (57.1%) were "hemolytic," 81 (33.1%) were "nonhemolytic," and 23 (9.8%) gave varied results. Only 12 (4.9%) of the strains produced spores that resisted boiling for 30 min or more. PMID- 16349699 TI - Microbial detoxification of aflatoxin. AB - Yeasts, molds, bacteria, actinomycetes, algae, and fungal spores were screened for their ability to degrade aflatoxin. Some molds and mold spores partially transformed aflatoxin B(1) to new fluorescing compounds. Only one of the bacteria, Flavobacterium (aurantiacum?) NRRL B-184, removed aflatoxin from solution. Both growing and resting cells of B-184 took up toxin irreversibly. Toxin-contaminated milk, oil, peanut butter, peanuts, and corn were completely detoxified, and contaminated soybean was partially detoxified by addition of B 184. Duckling assays showed that detoxification of aflatoxin solutions by B-184 was complete, with no new toxic products being formed. PMID- 16349700 TI - Virustat, a device for continuous production of viruses. AB - Methods for continuous production of viruses, and operation of the virustat, an apparatus in which such production was accomplished, were studied. Continuous production requires a separate continuous host growth chamber, such as the chemostat, and a multiunit virus growth chamber into which the virus-inoculated host cells are led. Successful continuous output of MS-2 and varphiX174 viruses, the latter in lysates, over periods of several days and at titers approximating those of batch lysates, was observed. Design problems include chamber sizes and flow rates, growth of resistant mutants within both virus and host growth chambers, clogging by lysis debris, and the phenomenon of self-inoculation. The latter represents virus growth in the first section of the chamber in excess of the washout rate, leading to lack of need for virus inoculation after an initial period. Use of the virustat for production and research purposes will require some attention to the formation of resistant bacterial colonies at pockets and surface sites of limited washout. With the virustat as a continuous virus production device, continuous purification methods are desirable. Research use of the virustat in continuous mutagenic population studies would require suppression of self-inoculation by use of many sections in the chamber, and improved servo control of host populations at low concentrations. PMID- 16349701 TI - Intracellular Loss of Potassium in Candida albicans After Exposure to Polyene Antifungal Antibiotics. AB - Eleven antifungal antibiotics, representing three broad macrolide classes, were studied in Candida albicans for their effect on growth and on the fate of intracellular K. Marked differences were observed among these antibiotics between their growth-inhibitory activities and their adverse effects on the integrity of the cellular membrane as evidenced by loss of K. Antibiotics most active in inhibiting growth of C. albicans were amphotericin B, trichomycin, candidin, candicidin (all heptaenes), and nystatin (a tetraene). In addition to those antibiotics, filipin and fungichromin also caused rapid leakage of K from yeast cells. Interestingly, fungichromin was the least active of the 11 antibiotics in inhibiting growth. Concentrations of rimocidin 10 times as great as those required for growth inhibition caused only a slight loss in intracellular K after 60 min. PMID- 16349702 TI - Effect of gamma Irradiation on the Microflora of Freshwater Fish: II. Generic Identification of Aerobic Bacteria from Yellow Perch Fillets. AB - Studies on the generic identification of bacteria isolated from nonirradiated and irradiated (0.3 and 0.6 Mrad) yellow perch fillets during the course of microbial spoilage have been conducted. After the enumeration and tabulation of macrocolonies on petri dish cultures obtained from fillets, isolates were examined and keyed out essentially according to modified morphological and biochemical protocols of Shewan. Identification was further confirmed through reference to Bergey's Manual. Data obtained from each isolate were coded and recorded on IBM cards to facilitate identification. Total aerobic microbial plate counts obtained from nonirradiated perch before storage ranged from 10 to 10 microorganisms per gram of fish. Organisms isolated from these fillets, in order of decreasing number, consisted of Achromobacter, Alcaligenes, Pseudomonas, Brevibacterium, Micrococcus, Flavobacterium, Bacillus, Sarcina, Microbacterium, Corynebacterium, yeasts, Lactobacillus, Vibrio, Aeromonas, and a few Proteus and Escherichia cells. During storage and as spoilage progressed, the flora shifted and the pseudomonads became predominant. Irradiation of fillets to 0.3 and 0.6 Mrad reduced the aforementioned flora to the Achromobacter-Alcaligenes group, which constituted the residual flora throughout fillet storage. PMID- 16349703 TI - Production of enterotoxin a. AB - A method for production of enterotoxin A in multiple liter lots is described. The medium contained 4% N-Z Amine NAK supplemented with 0.001% niacin and 0.00005% thiamine, and was adjusted to pH 6. The inoculated medium in lots of 400 to 600 ml, in 2-liter Erlenmeyer flasks, was incubated at 37 C for 24 hr on a gyrotory shaker at 280 rev/min. Production of 4 to 6 mug of enterotoxin A per ml occurred. PMID- 16349704 TI - Oxidative metabolism of dermatophytes. AB - A method for preparing young, actively respiring dermatophyte mycelia was obtained through the use of concentrated spore inocula and short growth periods in static culture. These hyphal elements were uniform in appearance, and vacuoles were absent. Concentrated mycelial suspensions were obtained which could be transferred easily and accurately. Glucose stimulated oxygen uptake in young mycelia which had been grown in a medium with low carbohydrate content. The level of endogenous respiration was affected by exogenous glucose only when this substrate stimulated oxygen uptake by less than 14%. Low nicotinamide adenine dinucleotide phosphate (NADP) dehydrogenase activity was noted in microconidia which have a low endogenous Q(o(2) ) value, whereas the activity of this enzyme was greater in macroconidia and mycelia which possess higher endogenous Q(o(2) ) values. Microsporum gypseum oxidizes 50% of exogenous glucose and assimilates the remainder. A large percentage of this substrate was assimilated into nitrogenous substances. PMID- 16349705 TI - Sensitive rapid detection method for viable bacterial cells. AB - A rapid sensitive method for the detection of viable bacterial cells is described in which P as inorganic orthophosphate is used to label the cells. Factors affecting the uptake of P by cells as well as the sensitivity of the method have been explored with suspensions of Aerobacter aerogenes. The uptake of PO(4) is dependent on several factors. Of various incubation media tested, one composed of 0.005 m KCl, 0.002 m MgSO(4) and 10 mg/ml of glucose was found to best stimulate the uptake of the tracer. Incubation time and temperature and level of isotope and of unlabeled P also affected uptake. Labeled cells were collected on a membrane filter for measurement of radioactivity. Under optimal conditions, as few as 23 viable cells per milliliter were detected in 1 hr with 95% confidence. PMID- 16349706 TI - Safe Self-contained Carbon Dioxide-Hydrogen Anaerobic System. AB - An anaerobic system is described which allows the microbiologist or hospital technician ease of operation not previously possible with other systems. Gas cylinders, vacuum pumps, valves, and gauges have been eliminated. A new anaerobic lid was developed that is fitted only with a snap-in rubber gasket, a double stainless-steel gauze flash arrestor, and a catalyst holder. The holder contains a palladium pellet catalyst which is active at room temperature and requires no heat for activation. This system was made specifically for use with a disposable hydrogen-carbon dioxide generator and a disposable methylene blue anaerobic indicator. In addition to ease of operation, this jar incorporates unique safety features which eliminate the possibility of a laboratory explosion. An oxygen free atmosphere composed primarily of nitrogen and carbon dioxide was quickly achieved within the jar to insure maximal growth. PMID- 16349707 TI - beta-Propiolactone Vapor Decontamination. AB - Although beta-propiolactone (BPL) is an effective vapor-phase decontaminant for enclosed areas, some problems have been encountered in its use. Adequate air circulation during BPL dissemination could eliminate most of these problems. It is recommended that, when decontaminating the ordinary building or laboratory, the amount of BPL sprayed be changed from the previously suggested 1 gal/16,000 cubic ft of space to 1 gal/25,000 cubic ft. The use of aqueous BPL solutions and thermal-type generators is not recommended. PMID- 16349708 TI - Clostridium tetani in a Metropolitan Area: A Limited Survey Incorporating a Simplified in Vitro Identification Test. AB - In a limited survey, three toxigenic and one nontoxigenic strains of Clostridium tetani were isolated from 18 environmental samples from metropolitan Boston. No C. tetani was found in 100 samples of human feces, 20 samples of dog feces, and two samples of horse feces. A simple modification of the halo precipitin test was studied in conjunction with the mouse lethality test for tetanus toxigenicity and was found to be a useful, although not a wholly definitive, technique. PMID- 16349710 TI - Isolation of thermophiles from broadleaf tobacco and effect of pure culture inoculation on cigar aroma and mildness. AB - Thermophilic members of the genus Bacillus isolated from fermented Connecticut broadleaf tobacco included eight strains of B. subtilis, five strains of B. coagulans, four strains of B. megaterium, and three strains of B. circulans. Some of these strains in pure single or mixed culture were employed to enrich the normal thermophile flora of "sweating" tobacco. Three strains of B. subtilis and one of B. circulans, either in single or multiple enrichment, caused the more rapid appearance of a pleasing aroma in Pennsylvania "Wrapper B" filler tobacco. These conclusions are based on subjective reactions of professional testers after numerous blindfold smoking tests. PMID- 16349709 TI - Anaerobic Production of Thiobacillus denitrificans for the Enzyme Rhodanese. AB - A method for the anaerobic growth of Thiobacillus denitrificans in a 140-liter (total capacity) stainless-steel culture vessel is described. As a result of controlling the pH value of cultures, and of ensuring that certain essential nutrients were in excess, cell yields approaching 700 mg (dry weight) per liter were obtained. These were over threefold higher than the best yields hitherto reported. The average rhodanese content of the cells from four cultures was 176,000 units per gram (dry weight). Adenosine-5'-phosphosulfate reductase (average content, 238 units per gram dry weight) and adenylate kinase (average content, 15,300 units per gram, dry weight) were also present. PMID- 16349711 TI - Copper sulfide precipitation by yeasts from Acid mine-waters. AB - Two strains of Rhodotorula and one of Trichosporon precipitated dissolved copper with H(2)S formed by reducing elemental sulfur with glucose. Iron stimulated this activity under certain conditions. In the case of Rhodotorula strain L, iron stimulated copper precipitation aerobically at a copper concentration of 18 but not 180 mug/ml. Anaerobically, the L strain required iron for precipitation of copper from a medium with 180 mug of copper per ml. Rhodotorula strain L was able to precipitate about five times as much copper anaerobically as aerobically. The precipitated copper was identified as copper sulfide, but its exact composition could not be ascertained. Iron was not precipitated by the H(2)S formed by any of the yeasts. Added as ferric iron, it was able to redissolve copper sulfide formed aerobically by Rhodotorula strain L from 18 but not 180 mug of copper per ml of medium. Since the yeasts were derived from acid mine-waters, their ability to precipitate copper may be of geomicrobial importance. PMID- 16349712 TI - Occurrence of zoogloea colonies and protozoans at different stages of sewage purification. AB - The presence of fingered branch-bearing Zoogloea has been noted on a number of occasions in the Baroda Sewage Disposal Works. Samples of raw sewage, the effluent from the continuous flow settling basin, the raw sludge, the floating scum in the settling basin, the final secondary digested sludge, and the supernatant liquid from the secondary digester were kept without any disturbance in 1-liter Pyrex glass beakers, which were loosely covered with petri dishes. Scum was formed on the surface within 48 hr in all the samples, and fingered Zoogloea colonies resembling the pure culture of Zoogloea ramigera reported by Crabtree et al. (5) were found in all except the final secondary digested sludge and the supernatant liquid from the secondary digester. It is not known whether the Zoogloea colonies discovered in the above cases are the same as or different from the typical Zoogloea ramigera of activated sludge, and whether they are slime-forming or flocculent types of bacteria. In any case, they seem to be different in their ecological status and in the nature of the accompanying protozoans from the typical Zoogloea ramigera. The reasons for the absence of zoogloeas in two of the samples are unknown. PMID- 16349713 TI - Comparison of methods for the isolation of salmonella from bone meal. AB - [This corrects the article on p. 711 in vol. 14.]. PMID- 16349714 TI - Fermentability of corn syrups with different dextrose equivalents added to various grape juices. AB - It was found that neither the enzymes of the grapes nor those of wine yeast Saccharomyces ellipsoideus strain 223 attacked the higher polysaccharides present in corn syrups. The alcohol yield of the corn syrups approached but did not quite reach theoretical assuming that all of the dextrose equivalent (DE) solids were fermented. Glucose, sucrose, and 95 DE corn syrup fermented at about the same rate and yielded comparable alcohol contents. At equal solids content, the 42 DE syrup fermented slower in most cases and yielded lower alcohol content than higher DE syrups. PMID- 16349715 TI - Glucose disappearance in biological treatment systems. AB - [This corrects the article on p. 861 in vol. 14.]. PMID- 16349716 TI - Importance of galacturonic Acid in controlling the retting of flax. AB - The empirical test was compared with the determination of pH, total acidity, and galacturonic acid (GA) as methods for establishing the termination time of the retting process in flax. The GA dosage was the best method for determining the end point of this process by distinctly dividing the phases of retting and over retting in the bacterial loosening of flax fiber. This end point is critical in the production of standard fibers for the weaving industry. PMID- 16349717 TI - Irradiation of Composts Used in the Propagation of Agaricus bisporus. AB - Unpasteurized horse manure composts were given radiation doses of 0.1 and 1.0 Mrad in an attempt to increase mushroom yields by eliminating competing microorganisms. Yields were enhanced by 0.1 Mrad but were lower than those obtained with heat-pasteurized composts. Composts subjected to 1.0 Mrad gave the lowest yields, probably because of the generation of ammonia and other toxic compounds. Only temporary changes in the predominant microflora could be detected as a result of ionizing radiations. PMID- 16349718 TI - Stimulation of yield in the cultivated mushroom by vegetable oils. AB - Supplementation of mushroom compost at spawning and at casing with various refined and crude seed oils resulted in 1 to 1.5 lb/ft increases in mushroom yield. Supplementation at casing with ground seeds or protein-oil combinations caused 2 to 2.5 lb/ft increases in mushroom yield. Further evidence is presented for a relationship between lipid metabolism and the initiation of fruiting in the cultivated mushroom, Agaricus bisporus (Lange) Sing. Preliminary results suggest the possible involvement of sterols in the fruiting stimulation. PMID- 16349719 TI - Spore Production by Bacillus stearothermophilus. AB - [This corrects the article on p. 690 in vol. 14.]. PMID- 16349720 TI - Aflatoxin Production by Aspergillus flavus as Related to Various Temperatures. AB - Two aflatoxin-producing isolates of Aspergillus flavus were grown for 5 days on Wort media at 2, 7, 13, 18, 24, 29, 35, 41, 46, and 52 C. Maximal production of aflatoxins occurred at 24 C. Maximal growth of A. flavus isolates occurred at 29 and 35 C. The ratio of the production of aflatoxin B(1) to aflatoxin G(1) varied with temperature. Aflatoxin production was not related to growth rate of A. flavus; one isolate at 41 C, at almost maximal growth of A. flavus, produced no aflatoxins. At 5 days, no aflatoxins were produced at temperatures lower than 18 C or higher than 35 C. Color of CHCl(3) extracts appeared to be directly correlated with aflatoxin concentrations. A. flavus isolates grown at 2, 7, and 41 C for 12 weeks produced no aflatoxins. At 13 C, both isolates produced aflatoxins in 3 weeks, and one isolate produced increasing amounts with time. The second isolate produced increasing amounts through 6 weeks, but at 12 weeks smaller amounts of aflatoxins were recovered than at 6 weeks. PMID- 16349721 TI - Growth Response of Lactobacillus brevis to Aeration and Organic Catalysts. AB - Under stationary and anaerobic conditions, greater cell yields of Lactobacillus brevis were obtained from autoclaved than from filter-sterilized glucose media. Fructose, tentatively identified as a product generated by the heating process, served as an excellent catalyst for inducing growth. The addition of micromolar quantities of pentoses or potential pentose precursors to the filter-sterilized medium was equally effective in stimulating growth. These organic catalysts were not essential for growth under aerobic conditions. Upon agitation, similar cell yields were obtained from the autoclaved and filter-sterilized media. The micromolar quantities of lactic acid produced per micromole of carbohydrate fermented appeared to be similar under aerobic and static conditions of incubation. The final concentration of acetic acid increased as the result of agitation. This increase in volatile acidity was accompanied by a significant decrease in ethyl alcohol production. The cell yield was increased nearly 50% under aerobic conditions. PMID- 16349722 TI - Utilization of Inorganic Nitrogen Compounds by Sphaerotilus natans Growing in a Continuous-Flow Apparatus. AB - Sphaerotilus natans was grown, attached, in a continuous-flow apparatus with inorganic nitrogen compounds (NH(4)Cl, NaNO(2), or NaNO(3)) as the only sources of nitrogen. The growth rate with NH(4)-containing medium did not differ from that with media containing glutamate or Casitone as the nitrogen source. PMID- 16349723 TI - Use of a methylene blue azide medium for isolation of enterococci. AB - A methylene blue azide medium (MBA), developed by Schaedler, Dubos, and Costello to isolate enterococci from the gastrointestinal tract of animals, was evaluated. This was done by comparing the isolation of enterococci from feces and saliva on the medium. Fifty-two catalase-negative, gram-positive cocci from human feces isolated from MBA were classified as enterococci. All strains grew in S F, 6.5% NaCl, and streptomycin broths, and all fermented mannitol. The isolates were provisionally subdivided into Streptococcus faecalis and S. faecium groups. S. faecalis-like strains fermented glycerol and pyruvate aerobically and produced acid in Snyder's medium (initial pH, 4.8). The S. faecium group fermented raffinose. Among all strains, several tests were variable. These included growth at 45 C, in 0.1% tellurite and in methylene blue milk. Three methods were employed to isolate and identify enterococci from the oral cavity. Direct streaking of MBA with saliva failed to produce any growth on the medium. Two other methods, with the use of various selective broths to promote the recovery of oral enterococci, failed to produce any bacteria capable of growing on MBA. The MBA-isolated fecal strains and oral viridans streptococci were generally indistinguishable on Mitis-Salivarius and K F agars. In experiments with fecal material, no gram-negative bacilli were found among the isolates selected. The MBA medium was judged as a high selectivity-low yield medium, and may provide a means of separating fecal and nonfecal enterococci. PMID- 16349724 TI - Florida citrus molasses as a fermentation substrate: I. Free amino acids in molasses from early and midseason varieties of citrus fruits during 2 years of production. AB - Samples of midseason Florida citrus molasses from 10 different manufacturing plants were studied during 2 years of production for suitability as fermentation substrates. They were analyzed for degrees Brix, pH, percentage of protein, and free amino acid content. The following ranges of values were obtained: degrees Brix, 65.4 to 73.0; pH, 4.2 to 6.4; and protein, 3.3 to 5.1%. Although the general characteristics of all samples were quite similar, the total free amino acid content and the number of amino acids differed from year to year and from plant to plant. The free amino acids consistently present were asparagine, aspartic acid, alanine, gamma-amino-butyric, glycine, glutamic acid, proline, serine, and threonine. Occasionally found were arginine, histidine, isoleucine, leucine, lysine, phenylalanine, tyrosine, and valine. An extremely high level of proline was detected in all samples. PMID- 16349725 TI - Degradation of Pure Aflatoxins by Tetrahymena pyriformis. AB - Tetrahymena pyriformis W with nutrients, ca. 22 x 10 cells, decreased the concentration of aflatoxin B(1) 58% in 24 hr and 67% in 48 hr. An unknown, bright blue fluorescent substance was produced, with intensity about one-half that of the unchanged B(1), with an R(f) of 0.52 compared with 0.59 for B(1) and 0.55 for B(2) on a thin-layer chromatography plate, and with an ultraviolet spectrum showing maxima of 253, 261, and 328 mmu. In a separate assay, the cells with nutrients did not degrade pure G(1). Starved, washed cells, ca. 11 x 10, decreased the concentration of B(1) 50% in 10 hr, 70% in 22 hr, and 75% in 30 hr, producing the same unknown component. Ethyl alcohol, 1.96% (v/v), decreased cell populations and size, but the cells remained actively motile in broth plus the alcohol for 96 hr. In 72 hr, neither toxin (ca. 2 ppm) in combination with ethyl alcohol had more inhibitory effect on cell numbers, with or without nutrients, than was produced by alcohol alone. Aflatoxin B(1) had no observed effect on the viability of the starved cells for 30 hr or on the nourished cells for 72 hr. There was no noticeable effect of G(1) on the starved cells in 30 hr or on the nourished cells in 48 hr. After 72 hr with G(1) plus nutrients, many of the cells were round with blisters, nonmotile, and apparently dead or dying. PMID- 16349726 TI - Enzyme patterns in the study of leptospira. AB - An analysis of intracellular and extracellular leptospiral enzymes was made by use of starch-gel electrophoresis with natural and synthetic substrates. Of 37 serotypes examined for extracellular exterase, all had activity of varying mobility and degree. All extracellular preparations were negative for catalase, phosphatase, and naphthylamidase. Intracellularly, five serotypes were examined, including Leptospira biflexa Patoc I, L. biflexa Waz, L. canicola Moulton, L. icterohaemorrhagiae RGA, and L. pomona S91. Among the enzymes detected by this electrophoretic technique were transaminase and catalase, confirming the results of previous investigators. Further, other enzymes heretofore unreported have been detected. These include esterases, phosphatases, lactic, malic, glutamic, succinic, alpha-glycerophosphate, and 6-phosphogluconic dehydrogenases, and a naphthylamidase. The presence of these enzymes suggests the existence of tricarboxylic acid, glycolytic, and pentose-related pathways in Leptospira. In addition, enzyme patterns show promise in leptospiral classification. PMID- 16349727 TI - Aerobic Heterotrophic Bacterial Populations of Sewage and Activated Sludge: III. Adaptation in a Synthetic Waste. AB - The nature and behavior of the microbial population developed on a synthetic waste containing salts and sorbitol are compared to that of a treatment-plant activated sludge. The population of the adapted sludge developed on the synthetic waste consisted approximately of only six kinds of bacteria. Experiments with radioactive sorbitol indicate that the microbial population developed on the synthetic waste showed the effects of repression by glucose on the metabolism of sorbitol; in contrast, the activated sludge from a plant treating primarily domestic waste was able to attack both substrates immediately and simultaneously. PMID- 16349728 TI - Immunofluorescence technique for the detection of salmonellae in various foods. AB - Salmonella species have been detected in nine food varieties by use of fluorescent antibodies without false-positive or false-negative results. Test antisera were specially prepared, commercially available, conjugated polyvalent O globulin absorbed with cultures of Escherichia coli and Citrobacter freundii, and polyvalent phase II H globulin antibodies. Use of this technique permits a decrease of 24 hr in time normally required for Salmonella detection when compared with cultural Salmonella recovery methods. PMID- 16349729 TI - Occurrence of an inhibitor of lactic Acid bacteria in green olives. AB - Green olives were found to contain an inhibitor(s) of several species of lactic acid bacteria usually associated with the Spanish-type brined olive fermentation. The inhibitor was demonstrated by the presence of inhibition zones surrounding tissue which had been cut from frozen olives and implanted in a seeded nutritive agar medium. Relative potencies of aqueous extracts of frozen olives were determined by a paper disc assay method. The Mission variety of olive contained the most inhibitor, and the Manzanillo and Ascolano, about 50 and 40% as much as the Mission variety, respectively. Sevillano and Barouni varieties contained comparatively little inhibitor. Effects of the inhibitor on growth rates of lactic acid bacteria were determined by adding various amounts of a concentrated aqueous extract of olives to a nutritive broth medium contained in screw-capped tubes. Of the four species of lactic acid bacteria tested, Leuconostoc mesenteroides was the most sensitive, and Lactobacillus plantarum was the least sensitive; Pediococcus cerevisiae and Lactobacillus brevis were intermediate in sensitivity. Extracts possessed a bactericidal property, as evidenced by their effect on L. mesenteroides. Sodium chloride, especially at concentrations of about 5% and higher, greatly increased the effectiveness of the inhibitor. The inhibitor was ethyl alcohol-soluble and was stable when heated at 100 C in aqueous solution. Potencies of extracts were reduced greatly by adjustment to pH 10, but no appreciable effect was noted by adjustment to pH 0.8. PMID- 16349730 TI - Dehydrogenase activity of pseudomonas species. AB - Single-strain cultures of Pseudomonas fragi, P. fluorescens, P. putrefaciens, and strains of two marine species, Pseudomonas type I and Pseudomonas type II, were found to be capable of reducing added acetaldehyde, proprionaldehyde, and butyraldehyde to the corresponding alcohols at 21 C. All species studied reduced propionaldehyde at 6 C. P. fragi, Pseudomonas type I, and Pseudomonas type II reduced butanone at 6 and 21 C. P. fragi and Pseudomonas type II reduced acetone at 21 C. Dehydrogenase activity was found in some cultures in which growth was not evident. Under aerobic conditions, a strain of P. fragi reduced added propionaldehyde to n-propanol quantitatively within 36 hr at 21 C. PMID- 16349731 TI - Observations on the Occurrence, Distribution, and Seasonal Incidence of Blue green Algal Viruses. AB - Phycovirus populations were found in 11 of the 12 waste stabilization ponds studied. These populations were comprised solely of blue-green algal (BGA) viruses. Two virus types were observed, one of which was related to the previously reported LPP-1 virus. The incidence and magnitude of the LPP group indicated that several of the ponds supported well-established BGA virus populations of this type. Counts as high as 270 plaque-forming units/ml were noted; however, marked differences in the nature and magnitude of these BGA viruses were apparent even in geographically related ponds of similar design. Of the algal strains found dominant in these ponds, none was of the type reported susceptible to the LPP viruses. PMID- 16349732 TI - Rate of isolated hemicellulose degradation and utilization by pure cultures of rumen bacteria. AB - Rate studies on the utilization or degradation (or both) of isolated hemicelluloses were conducted with six strains of rumen cellulolytic bacteria. Utilization was estimated by total pentose loss, and degradation values were based on solubilization of the hemicellulose in acidified 80% ethyl alcohol. With the various strains of ruminococci, degradation of flax and fescue grass hemicellulose was near the maximum within the first 12 hr of incubation. However, where applicable, the rates of utilization were considerably slower. Both degradation and utilization of corn hull hemicellulose occurred at much slower rates than observed with the other two substrates. With flax and fescue grass hemicellulose, the rates of degradation did not appear to be influenced by the organism's ability, or inability, to utilize the substrate as an energy source. The rates and extent of isolated hemicellulose degradation and utilization were compared between the cellulolytic ruminococci and three strains of bacteria isolated from the rumen with a xylan medium. Similar values were obtained with both types of bacteria. These observations would suggest that the cellulolytic ruminococci may be important in the overall fermentation of forage hemicelluloses in the rumen. The acidified 80% ethyl alcohol supernatant fluids, obtained from fermentations of isolated fescue grass hemicellulose by two strains of Ruminococcus flavefaciens, of which only one was able eventually to utilize the substrate, were investigated by thin-layer chromatography. Results indicated that soluble oligosaccharides were produced, which were observed to disappear gradually with time in fermentations with the utilizing strain and to accumulate in fermentations with the nonutilizing strain. Examination of the acidified 80% ethyl alcohol-insoluble residue hydrolysates, obtained from fermentations with the utilizing strain, revealed that the concentration of all the constituent sugars decreased uniformly. PMID- 16349733 TI - Respiratory Activity of the Mycelium of Eremothecium ashbyii. AB - The respiratory activity of the mycelium of Eremothecium ashbyii from submerged cultures was manometrically determined at different stages of its development and the results were statistically analyzed. The experiments were performed in a manner designed to diminish the endogenous respiration without affecting the response to the addition of an exogenous substrate. Lactose was the carbohydrate tested that produced the lowest oxygen consumption. The oxidation of maltose, which was high at 24 hr, decreased by more than 50% at 48 and 55 hr. Glucose and sucrose were actively oxidized by mycelium of three ages. From the intermediates of carbohydrate metabolism, 24-hr mycelium did not produce oxygen consumption with malate, lactate, citrate, fumarate, and alpha-ketoglutarate. At 48 hr, mycelium did not oxidize either lactate or citrate; 55-hr mycelium showed oxygen consumption with all intermediates tested. Acetate and pyruvate always produced high oxygen consumption. Ethyl alcohol produced high oxygen consumption with mycelium of all tested ages. PMID- 16349734 TI - Mechanism of Thermal Injury in Staphylococcus aureus: I. Relationship Between Viability and Leakage. AB - The viability of and leakage from suspensions of Staphylococcus aureus stored in the temperature range 3 to 60 C were determined. There was a direct relationship, up to 50 C, between death and leakage, both of which increased with increasing temperature. At temperatures above 50 C, there was a different pattern of response, the total leakage being less at 60 C than at 50 C, although there was greater membrane damage at the higher temperature. Sucrose, 1 m, almost completely prevented leakage at 37 and 50 C but not at 60 C. PMID- 16349735 TI - Production of Lysergic Acid Derivatives in Submerged Culture: IV. Inorganic Nutrition Studies with Claviceps paspali. AB - The inorganic requirements for growth and alkaloid production by submerged cultures of a lysergic acid alkaloid-producing isolate of Claviceps paspali were determined in a defined medium. The requirement for peak growth was essentially the same as the requirement for maximal alkaloid production in the case of potassium and magnesium. With phosphorus, however, maximal growth was reached at lower levels than those required for maximal alkaloid production. Sulfur was required in smaller amounts for peak alkaloid production than for maximal growth. In the case of iron and zinc, the requirement for maximal alkaloid production was greater than for growth. Manganese and copper could be sufficiently depleted to demonstrate their essentiality for alkaloid production, but their requirements for growth could not be demonstrated. PMID- 16349736 TI - Effect of Growth Rate on the Synthesis of Penicillin by Penicillium chrysogenum in Batch and Chemostat Cultures. AB - The kinetics of penicillin production by Penicillium chrysogenum Wis 54-1255 in a glucose-limited chemostat and in batch cultures are reported. The specific production rate of penicillin, q(pen) (units per milligram of dry weight per hour) was independent of specific growth rate over the range 0.014 to 0.086 hr. Growth was stopped by restricting the glucose supply to the "maintenance ration," that is, the glucose requirement of the organism at zero growth rate with all other nutrients in excess. Under such conditions, the organism dry weight remained constant, but the q(pen) fell approximately linearly to zero at a rate inversely related to the previous growth rate. Glucose supplied in excess of the maintenance ration inhibited the decay of q(pen). At a critical growth rate between 0.009 and 0.014 hr, the decay was completely inhibited. Quantitative expressions for the q(pen) of growing and nongrowing cultures were derived and used to predict the steady-state concentrations of penicillin accumulating in one and two-stage continuous processes. A rational explanation of the kinetics of penicillin accumulation in batch cultures is given, relating the rate of penicillin synthesis to growth rate. It is concluded that an important role of corn steep liquor (CSL), a heterogeneous carbon and nitrogen source commonly used in penicillin production media, is the provision of substrates which allow a high concentration of mold to be reached before the growth rate falls below the critical value. CSL had no significant effect on q(pen). PMID- 16349737 TI - Inhibition of microorganisms by topical anesthetics. AB - The effect of various topical anesthetics and their preservatives on the growth of Pseudomonas aeruginosa, Staphyloccoccus albus, and Candida albicans was investigated. The topical anesthetics were proparacaine HCl, tetracaine HCl, cocaine HCl, and benoxinate HCl. The preservatives were chlorobutanol and butyl p hydroxybenzoate. Proparacaine inhibited C. albicans but not P. aeruginosa or S. albus. All three test organisms were inhibited to varying degrees by tetracaine, benoxinate, cocaine, chlorobutanol, and butyl p-hydroxybenzoate. PMID- 16349738 TI - Effect of Water Vapor on Lyophilized Serratia marcescens and Escherichia coli. AB - Dried Serratia marcescens ATTC 14014 and Escherichia coli ATTC 4157 cells were exposed to various partial pressures of purified water vapor. The colony-forming ability of the S. marcescens was unimpaired when the dried organisms were stored in water-vapor atmosphere such that P/P(0) < 0.55 or P/P(0) = 1.0 (where P is the pressure of the water vapor in contact with the organisms, and P(0) is vapor pressure of pure water at 25 C). During storage under water-vapor atmospheres with P/P(0) between 0.6 and 1.0, the colony-forming ability of the dried S. marcescens was destroyed. The inactivation by water vapor followed the expression - ln N/N(0) = Kt, where N(0) and N are the number of viable organisms before and after exposure, respectively, t is time, and K is a pseudo constant which is dependent upon the partial pressure of the water vapor at 25 C. Similar results were obtained with dried E. coli. The addition of solutes to the suspending media before freeze-drying was found to influence the stability of the organisms during exposure to water vapor. PMID- 16349739 TI - Spherical Lactic Acid-producing Bacteria of Southern-grown Raw and Processed Vegetables. AB - The frequency and levels of population of the spherical lactic acid-producing bacteria were determined on raw and processed yellow summer and zucchini squash, a variety of greens, green beans, okra, southern peas, and butter and lima beans, and on fresh cucumbers and corn flowers. Six taxa occurred consistently: Leuconostoc mesenteroides, yellow-pigmented streptococci, Streptococcus faecium, Aerococcus viridans, and S. faecalis and S. faecalis var. liquefaciens. The same taxa occurred with the same order of frequency on processed, frozen vegetables, but with a marked decrease in the occurrence of S. faecalis var. liquefaciens. S. lactis, S. cremoris, S. equinus, S. bovis, and pediococci were isolated infrequently. No other member of the viridans group of the streptococci and no member of the pyogenic group was isolated. Approximately 88% of the cultures were identified. Total counts of the lactic-acid-producing bacteria rarely exceeded 10 per gram of sample, and there was a reduction by 90% during the second year of study, probably because of drought. Only one bacterial species was found on 40% of the raw and 34% of the processed vegetable samples. Two or more species or taxa were present on the remainder of 153 raw and 56 processed vegetable samples. A. viridans was present on squash, greens, okra, and southern peas, and its frequency of occurrence on vegetables suggests that plants are its natural habitat. PMID- 16349740 TI - Bacteriological Survey of the Frozen Prepared Foods Industry: III. Potato Products. AB - During sanitation-inspections of 29 potato-processing firms, 2,544 finished product units and 1,654 samples were collected and analyzed bacteriologically. The results of the bacteriological examination of finished French fries, fried potato cylinders, and dehydrated potatoes usually did not reveal the conditions of cleanliness under which they were produced. This was most likely due to the lethal effect of the terminal fry to which the French fries and potato cylinders are subjected, and because of the lethal effect of the dehydration temperatures used in processing the dehydrated potato products. However, as in most food processing firms, line samples collected at each processing step reflect sanitary conditions and provide bacteriological support of inspectional evidence of plant insanitation. Most of the frozen stuffed baked potatoes examined were produced under poor sanitary conditions. But because this product is usually processed while hot, bacteriological examination of the finished product did not usually reveal the conditions of cleanliness under which they were produced. Again, inspectional observations were necessary for a full evaluation of the conditions of production. In contrast, frozen potato patties and frozen hash brown potatoes were more likely to reveal the conditions of sanitation under which they were produced as evidenced by the varying bacterial counts. PMID- 16349741 TI - Growth yields of bacteria on selected organic compounds. AB - Cell yields were determined for two bacterial soil isolants grown aerobically in minimal media on a variety of synthetic organic compounds. 1-Dodecanol, benzoic acid, phenylacetic acid, phenylglyoxylic acid, and diethylene, triethylene, and tetraethylene glycols were tested. Two "biochemicals," succinate and acetate, were also tested for comparison. Yields were calculated on the basis of grams of cells obtained per mole of substrate utilized, gram atom of carbon utilized, mole of oxygen consumed, and equivalent of "available electrons" in the substrates. This latter value appears to be nearly constant at 3 g of cells per equivalent of "available electrons." Yields predicted on this basis for other bacteria and for yeasts on other substrates are in fair agreement with reported values. PMID- 16349742 TI - Continuous Propagation of Trichosporon cutaneum in Cheese Whey. AB - Trichosporon cutaneum, a nonfermenting yeast, was used to convert cheese whey lactose into microbial cell material. The doubling time for this organism in a laboratory-scale continuous propagator was 2 hr in a whey medium fortified with ammonium sulfate and corn steep liquor. Cellular growth and efficiency of conversion of lactose to cell material was higher than with Saccharomyces fragilis. When grown in whey, the nitrogen content of T. cutaneum was 3.5% and the distribution of amino acids per gram of cell protein was similar to that of commercial food yeasts. PMID- 16349743 TI - Use of ultrasonic energy in assessing microbial contamination on surfaces. AB - Ultrasonic tanks were evaluated for their ability to remove viable microorganisms from various surfaces for subsequent enumeration. Test surfaces were polished stainless steel, smooth glass, frosted glass, and electronic components. The position of contaminated surfaces in relation to the ultrasonic energy source, distance of the ultrasonic source from the test surfaces, and temperature of the rinse fluid were some of the factors which influenced recovery. Experimental systems included both naturally occurring microbial contamination and artificial contamination with spores of Bacillus subtilis var. niger. The results showed that ultrasonic energy was more reliable and efficient than mechanical agitation for recovering surface contaminants. Conditions which increased the number and percentage of microorganisms recovered by ultrasonic energy were: using a cold rinse fluid, placing the sample bottle on the bottom of the ultrasonic tank, and facing the contaminated surfaces toward the energy source. It was also demonstrated that ultrasonic energy could be effectively used for eluting microorganisms from cotton swabs. PMID- 16349744 TI - Microbial penetration and utilization of organic aircraft fuel-tank coatings. AB - Microorganisms have been found as contaminants in various types of aircraft fuel tanks. Their presence introduces problems in the operation of the aircraft, including destruction of components such as the organic coatings used as protective linings in the fuel tanks. Microbial penetration and utilization of the currently used organic coatings, EC 776, DV 1180, PR 1560, and DeSoto 1080, were determined by changes in electrical resistances of the coatings; mycelial weight changes; growth counts of the bacteria; and manometric determinations on Pseudomonas aeruginosa (GD-FW B-25) and Cladosporium resinae (QMC-7998). The results indicate EC 776 and DV 1180 to be less resistant to microbial degradation than the other coatings. Organic coatings, serving as a source of nutrition, would be conducive to population buildups in aircraft fuel tanks. PMID- 16349745 TI - Chemical composition of variants of aerobic actinomycetes. AB - It has been shown previously that aerobic actinomycetes can be separated into four main groups on the basis of their cell wall composition. Six representatives of aerobic actinomycetes (Nocardia asteroides and Micropolyspora brevicatena, cell wall type IV; N. madurae, Microbispora rosea, cell wall type III; Actinoplanes sp., cell wall type II; Streptomyces griseus, cell wall type I) were subjected to selecting agents which permitted the isolation of stable variants morphologically different from the parent strain. Whole cell analyses of 134 substrains from the six parents revealed no significant change in the isomeric form of diaminopimelic acid or in sugar constituents. Analyses of cell wall preparations from 52 of these did not reveal any change in the diagnostic constituents of their murein or polysaccharides. PMID- 16349746 TI - Seasonal Variations in Survival of Indicator Bacteria in Soil and Their Contribution to Storm-water Pollution. AB - Survival of a fecal coliform (Escherichia coli) and a fecal streptococcus (Streptococcus faecalis var. liquifaciens) was studied through several years at shaded and exposed outdoor soil plots. Death rates for both organisms were calculated for the different seasons at both sites. The 90% reduction times for the fecal coliform ranged from 3.3 days in summer to 13.4 days in autumn. For the fecal streptococcus, 90% reduction times were from 2.7 days in summer to 20.1 days in winter. During summer, the fecal coliform survived slightly longer than the fecal streptococcus; during autumn, survival was the same; and in spring and winter the fecal streptococcus survived much longer than the fecal coliform. Both organisms were isolated from storm-water runoff collected below a sampling site when counts were sufficiently high in soil. Isolation was more frequent during prolonged rains, lasting up to 10 days, than during short rain storms. There was evidence of aftergrowth of nonfecal coliforms in the soil as a result of temperature and rainfall variations. Such aftergrowth may contribute to variations in bacterial count of storm-water runoff which have no relation to the sanitary history of the drainage area. PMID- 16349747 TI - Effect of Microwaves on Escherichia coli and Bacillus subtilis. AB - Suspensions of Escherichia coli and Bacillus subtilis spores were exposed to conventional thermal and microwave energy at 2,450 MHz. The degrees of inactivation by the different energy sources were compared quantitatively. During the transient heating period by microwave energy, approximately a 6 log cycle reduction in viability was encountered for E. coli. This reduction was nearly identical to what is expected for the same time-temperature exposure to conventional heating. Heating of B. subtilis spores by conventional and microwave energy was also carried out at 100 C, in ice and for transient heating. The degree of inactivation by microwave energy was again identical to that by conventional heating. In conclusion, inactivation of E. coli and B. subtilis by exposure to microwaves is solely due to the thermal energy, and there is no per se effect of microwaves. PMID- 16349748 TI - Antimicrobial performance of tri- and tetrachloro-4-(methylsulfonyl) pyridines as industrial preservatives. AB - Tests show that trichloro- and tetrachloro-4-(methylsulfonyl) pyridines have a wide spectrum of activity against microorganisms causing deterioration in industrial systems. They are particularly active as fungistats and are more effective against gram-positive than against gram-negative bacteria. Their fungistatic activity and persistence have led to outstanding performance as mildewcides in exterior paints. In addition, we report the performance of these compounds as preservatives in the following products: asphalt, calking compositions, joint cements, paper, plastics, soluble oils (cutting fluid emulsions), and textiles. PMID- 16349749 TI - Serological identification of enterotoxigenic staphylococci from cheese. AB - Single and double gel-diffusion techniques were employed to examine serologically coagulase-positive staphylococci from cheese for enterotoxigenicity. Supernatant fluid from sac cultures was examined for enterotoxins A and B. The results indicated that 9 of 155 cultures from market cheese and 7 of 77 cultures from food-poisoning cheese produced enterotoxin A, and that none of the cultures produced detectable levels of enterotoxin B. Results of serological tests were confirmed by intravenous injection of cats. PMID- 16349750 TI - Recovery of microorganisms shed by humans into a sterilized environment. AB - An appartus and technique for quantitative comparison of the aerobic bacterial flora disseminated by human subjects has been developed. Dissemination from three healthy subjects was studied weekly for 3 weeks. Viable particles recovered ranged from 100,000 for one subject during a 30-min period to 620,000 for another subject during a 10-min period. One of the three subjects showed appreciably less variation in numbers of organisms shed than did the other two subjects. When the subjects were examined on consecutive days while wearing sterilized clothing, total particles recovered were reduced and variations in recoveries from run to run were slightly lessened. Three consistent nasal carriers of S. aureus were measured for dissemination. No viable Staphylococcus aureus was recovered from two of the carriers. However, 460,000 typable S. aureus particles were recovered during a 60-min period from the third carrier. PMID- 16349751 TI - Metabolism of 4-chloro-2-methylphenoxyacetic Acid by soil bacteria. AB - A microorganism capable of degrading 4-chloro-2-methylphenoxyacetic acid (MCPA) was isolated from soil and identified as Flavobacterium peregrinum. All of the chlorine of MCPA was released as chloride, and the carboxyl-carbon was converted to volatile products by growing cultures of the bacterium, but a phenol accumulated in the medium. The phenol was identified as 4-chloro-2-methylphenol on the basis of its gas chromatographic and infrared characteristics. Extracts of cells of F. peregrinum and of a phenoxyacetate-metabolizing Arthrobacter sp. dehalogenated MCPA and several catechols but not 4-chloro-2-methylanisole. The Arthrobacter sp. cell extract was fractionated, and an enzyme preparation was obtained which catalyzed the conversion of MCPA to 4-chloro-2-methylphenol. The latter compound was not metabolized unless reduced nicotinamide adenine dinucleotide phosphate was added to the fractionated extract. The phenol in turn was apparently oxidized to a catechol by components of the enzyme preparation. PMID- 16349752 TI - Quantitative analyses of certain enteric bacteria and bacterial extracts: I. Standardization and sonic disruption of eight enteric bacterial species, each at five population levels. AB - Standardized individual preparations of five population levels of eight enteric organisms [Escherichia coli (O4:H3), E. coli (O111:B4:H12), Salmonella enteritidis, S. paratyphi B, S. typhimurium, Shigella boydii, S. dysenteriae, and S. sonnei) were prepared. Dry weights, calculated mean cell weight, and nitrogen content of bacterial suspensions before, and of supernatant fluids after, ultrasonic disruption are tabulated. Percentages of disruption, estimated from nitrogen concentration ratios of the suspensions and supernatant fluids, are given. These data are presented as guidelines for the preparation of bacterial extracts prior to precipitin analyses. PMID- 16349753 TI - Quantitative Analyses of Certain Enteric Bacteria and Bacterial Extracts: II. Discrimination of Sonic Extracts by Interfacial Densitometry of Precipitin Systems. AB - Bacterial extracts prepared by ultrasonic disruption were reacted with both narrow- and broad-spectrum reference (homologous) and cross-reacting (heterologous) precipitins produced in rabbits. Quantitation of the reaction was obtained by densitometry of the antigen-antibody interface. Comparisons were made of sonic extracts from various starting populations all equated to the same nitrogen concentrations, and of various nitrogen levels derived from five bacterial population levels prepared separately. Sources of error are probed to show under what circumstances cross-reactions would be of greater magnitude than reference ones. The feasibility was shown of using quantitative densitometry of the interface combined with broadly reacting precipitins to identify bacteria on an intergeneric and interspecies scale. Problems associated with the use of absorbed or monospecific precipitins are explained. PMID- 16349754 TI - Production of hyaluronic Acid by a streptococcal strain in batch culture. AB - A simple method for the production and preparation of hyaluronic acid (HA) is described. Three media were tested, which all supported relatively good yields of cells, but only in one of them were appreciable amounts of HA formed. In this medium, HA was produced during the logarithmic growth phase and showed a molecular weight of 7.3 x 10. HA prepared from 24-hr cultures of the same organism had a molecular weight of only 2.6 x 10. The kinetics of HA production in batch culture were studied, as well as of cell production and glucose consumption. PMID- 16349755 TI - Friend Leukemogenic Virus-neutralizing Antibody from Mouse Ascitic Fluid. AB - Ascitic fluid antibody produced in C57/B1 mice immunized with Friend leukemogenic virus exhibited potent neutralizing activity. In vitro neutralization tests revealed that a mean neutralization index of 3.0 was achieved, and it was shown by sucrose gradient ultracentrifugation that this antibody resembled the 7S type. A mean yield of 6.7 ml of fluid per mouse per weekly paracentesis was obtained over an 8-week period. The ascitic fluid antibody to Friend virus was also active in vivo. Mice given antibody 3, 5, 7, and 9 days after infection with Friend virus did not develop the splenomegaly characteristic of Friend disease. PMID- 16349756 TI - Protozoan, bacterial, and volatile Fatty Acid changes associated with feeding tylosin. AB - Tylosin was fed to two of six wethers for 79 days, to a second two for only 28 days, and not at all to a third pair (controls). The addition of tylosin to the daily feed resulted in a rapid twofold increase in protozoal concentration and a change in the composition or characteristics, or both, of the bacterial population. The results indicate that the bacterial population was modified to the extent of about 80%. Total acid concentrations were initially depressed but appeared to be greater than those in control animals at the termination of the experiment. Deletion of tylosin from the ration resulted in a rapid decrease in protozoal concentrations, whereas changes in the bacterial population did not occur for a further 30 days. PMID- 16349757 TI - Effect of lipid materials on heat resistance of bacterial spores. AB - The apparent heat resistance of spores of Bacillus megaterium, B. subtilis, B. cereus, B. stearothermophilus, and Clostridium botulinum type E in lipids was investigated and compared with the resistance of the spores in phosphate buffer solution. The most pronounced increase in heat resistance was noted for B. subtilis and C. botulinum type E, the increase varying with the type of lipid used. A high water content of the lipids used as heating menstruum lowered the heat resistance of the spores. Possible explanations for the high heat resistance of spores in lipids are discussed. PMID- 16349758 TI - Degradation of Glucose by Proliferating Cells of Desulfotomaculum nigrificans. AB - Desulfotomaculum nigrificans degraded glucose to acetate, ethyl alcohol, and carbon dioxide. By use of C-glucose labeled at different carbon atoms, two pathways of glucose metabolism were detected. They were the Embden-Meyerhof and the Entner-Doudoroff schemes. Because the observed quantities of acetate and carbon dioxide, arising from glucose, were greater than the expected theoretical values, individual fermentations were conducted with 15 uniformly labeled C-amino acids. The results indicated that amino acids, supplied by the yeast extract or peptone in the fermentation medium, also contributed to the formation of acetate and carbon dioxide. PMID- 16349759 TI - Microbial conversion of acetanilide to 2'-hydroxyacetanilide and 4' hydroxyacetanilide. AB - Approximately 700 cultures of various types were examined for their ability to hydroxylate acetanilide. The major product formed by unidentified Streptomyces species RJTS-539 was identified as 4'-hydroxyacetanilide (N-acetyl-p aminophenol). This culture gave a peak yield of 405 mg per liter from 1,000 mg of acetanilide per liter. Considerably lower yields of 4'-hydroxyacetanilide were isolated from S. cinnamoneus NRRLB-1285. The major conversion product of acetanilide formed by Amanita muscaria F-6 was identified as 2' hydroxyacetanilide, with a peak yield of 433 mg per liter from 1,000 mg per liter of substrate. A small amount of 4'-hydroxyacetanilide was also formed. Six other Streptomyces cultures formed small amounts of one or two products identical or similar to 2'-hydroxyacetanilide or 4'-hydroxyacetanilide as determined by thin layer chromatography and ultraviolet spectra. PMID- 16349761 TI - Control of Pleuropneumonia-like Organisms in Cell Culture. AB - Mammalian cell culture systems were maintained free of mycoplasmas by using a 3 day agar plate test as a weekly routine to monitor the conditions of the cells. If contaminated cell cultures were found, they were discarded and replaced from a pleuropneumonia-like organism (PPLO)-free cell bank. PPLO-free lines were established by treatment with various antibiotics. The KB cell line was freed of mycoplasmas by treatment for 1 week with a mixture of chlortetracycline, kanamycin, and chloramphenicol. L-929 cells were cleared of contamination with either spectinomycin or tylosin, and a synovial cell line was cleared with lincomycin or tylosin. Each cell line, after eradication of the contaminant, was stored in liquid nitrogen. A number of agents were tested to determine minimal inhibitory concentration against three known and three unidentified mycoplasmas. Chlortetracycline and tetracycline were found to be highly active against all strains, whereas tylosin, spectinomycin, and lincomycin, though less active, were equally useful because of their low toxicity against cells. Kanamycin was highly active against three strains, but inactive at high levels against the KB cell contaminants. A disc plate test was used to check isolated cell contaminants for sensitivity to various agents. PMID- 16349760 TI - Comparison of Fluorescent Antibody with Cultural Technique for Isolation of Enteropathogenic Escherichia coli from Swine. AB - In an investigation of hogs as possible reservoirs of human strains of enteropathogenic Escherichia coli (EEC), 92 six-month-old grain- and garbage-fed hogs were examined on the farm and again at the packing plant. Of the 331 specimens obtained by swabbing the rectum, cecum, and edible meat carcass of these hogs, 125 were presumptively positive for EEC when screened by the fluorescent-antibody (FA) technique. These "presumptive positive" specimens then underwent extensive bacteriological examination and complete serological typing. The FA technique proved to be an easier, simpler, and more economical procedure than culture when a large number of specimens were examined for possible EEC serogroups. It was found especially valuable for identification of multiple serogroups of EEC within a single specimen. It also appeared to be more sensitive than cultural examination, since results were not dependent on the presence of large numbers of organisms in the specimen, or even on their viability. However, the FA technique was found to be less specific than culture because of cross reactivity with antigenically related Enterobacteriaceae when fluorescein-labeled antisera were used. Therefore, any specimen found positive on FA examination should be considered as presumptive positive until confirmed by bacteriological examination and complete serological study. PMID- 16349762 TI - Bacteriophage aerosol challenge of installed air contamination control systems. AB - The effectiveness of installed air contamination control systems for removal of airborne microorganisms was determined. Seven separate systems were challenged with aerosolized T1 bacteriophage. Air samples were collected with Andersen samplers containing agar plates that had been swabbed with susceptible Escherichia coli. The advantages of using bacteriophage for testing air-handling systems are discussed. The air systems employed both filtration and ultraviolet irradiation and air-flow rates varied from 600 to 18,000 ft/min. Reduction rates of airborne bacteriophage passing through the various air systems ranged from 99.8 to 99.99%. PMID- 16349763 TI - Rumen microorganisms in buffalo from southern utah. AB - Rumen microbial populations from buffalo (Bison bison bison Linn.) in southern Utah were identified on the basis of their morphology and staining characteristics. The rumen bacteria and ciliate protozoa were similar in number and kind to those found in domestic livestock. PMID- 16349764 TI - Photodynamic inactivation of newcastle disease virus with acridine orange. AB - Newcastle disease virus in allantoamnionic fluid was photosensitized with acridine orange, and then photodynamically inactivated. The inactivation rate was directly proportional to light intensity and to dye concentrations up to a level of approximately 100 to 200 mug/ml. Inactivation occurred at approximately an exponential rate for the first 4.5 log(10) units of virus, and then continued at a decreasing rate. PMID- 16349765 TI - Inactivation of Venezuelan Equine Encephalomyelitis Virus by gamma-Radiation. AB - Exposure of Venezuelan equine encephalomyelitis (VEE) virus (at -70 C) to 6 x 10 r gamma-radiation (Co) resulted in loss of lethality for young adult mice and guinea pigs, and loss of capacity to produce plaques or cytopathic effects in tissue culture. The suckling mouse was more sensitive for detecting live virus in radiated suspensions than was the adult mouse or guinea pig. Live virus was demonstrable in preparations exposed to 6 x 10 r but not in suspensions exposed to 8 x 10 r and more. The rate of inactivation of VEE virus by gamma-radiation was an exponential function of the dosage. PMID- 16349766 TI - Propagation of Rhinovirus on WI-38 Cell Monolayers in Rolling Bottles. AB - Techniques are described for the propagation of rhinoviruses on WI-38 monolayers in rolling bottles. High yields of viruses were obtained, as indicated by infectivity titers and electron microscopy. When crude harvests were subjected to low-speed centrifugation and then filtered through a 0.45-mu membrane filter, little or no loss in infectivity titer was observed. However, electron microscopic examination indicated that the concentration of viral physical particles was reduced below detectable levels after filtration. The guinea pig potency test on the lot of unfiltered rhinovirus 14 vaccine prepared in rolling bottles indicated that this vaccine stimulated higher reciprocal serum neutralizing titers than a rhinovirus 14 vaccine prepared in stationary monolayers. PMID- 16349767 TI - Effect of environment on staphylococcal lesions in mice. AB - The influence of reduced barometric pressure on the development and healing of staphylococcal skin lesions in mice was investigated by exposing groups of animals to the test environment before, after, or before and after subcutaneous inoculation with 3.5 x 10 colony-forming units of a phage type 80 strain of Staphylococcus aureus. Similarly infected control animals were not exposed to the experimental environment. The results indicate that the lesions which developed in mice exposed to the test environment prior to infectious challenge were larger and healed at a slower rate than those in mice maintained at ground level before infection. Exposure after inoculation produced no demonstrable effect in the size or healing rate of the experimental lesions. PMID- 16349768 TI - Growth of salmonella in chickens' yolk sacs and its relationship to pathogenicity. AB - Newly hatched chicks were inoculated in the yolk sacs with standardized suspensions of Salmonella anatum, S. heidelberg, or S. infantis. At intervals between 3 and 48 hr postinoculation, chicks from each group were sacrificed, the average number of viable cells per yolk sac was determined, and liver tissue from each chick was examined for Salmonella. Growth patterns of the three species were almost identical when each chick was inoculated with about 3.5 million cells, but S. heidelberg was recovered more frequently from the liver, and caused a much higher percentage of mortality than did either S. anatum or S. infantis. When 100 fold dilutions of the suspension of S. heidelberg were used, mortality and recovery rates of the bacterium from the liver were directly related to the number of cells injected. The logarithmic growth phase was lengthened as the number of cells in the inocula was decreased; consequently, there was little difference in the average number of S. heidelberg cells per yolk sac at 36 or 48 hr postinoculation regardless of number of cells injected. Results of these trials indicated that factors other than rate of multiplication in the yolk sac are responsible for observed differences between Salmonella species in degree of pathogenicity for baby chicks. PMID- 16349769 TI - Cell yields of bacteria grown on methane. AB - Several mixed cultures of methane-oxidizing bacteria have been isolated. Among them, culture HR (consisting of two gram-negative rods, one 0.5 x 1.0 mu, the other 0.8 x 2 to 3 mu) was found to be the fastest-growing and to give the highest yields. Optimal conditions for rapid growth and high cell yields from methane were found to be: 30 C, NH(4) as nitrogen source, and pH 6.5. Requirements for CO(2) and Cu were observed. Under these conditions, generation times of approximately 3 hr and cell yields from methane between 65 and 70% could be attained. Culture HR can utilize methane, methanol, ethyl alcohol, 1-propanol, n-butyl alcohol, and glucose, but not propane, for growth. Yeast and beef extracts are inhibitory. Carbon balances demonstrate that few if any products other than cells and CO(2) are produced from methane under the growth conditions used. Cell analyses for carbon, hydrogen, nitrogen, and amino acid content of culture HR were also made. PMID- 16349770 TI - Effects of Supplemental Calcium or Calcium-binding Agents on Staphylococcal Bacteriophage Proliferation in Skim Milk. AB - Additions of 0.0005 N calcium borogluconate to Trypticase Soy Broth (TSB) produced an increase in phage titer about 1 million-fold, whereas its addition to skim milk resulted in about a 100-fold decrease in the maximal titer. Supplemental calcium had a stimulatory influence on bacterial growth in TSB but not in skim milk. Studies were made of the effect of binding of calcium of skim milk on the proliferation of staphylococcal bacteriophage. Sequestering the calcium with 2% phosphate mixture inactivated the phages without affecting the bacterial growth. However, chelation of calcium by 0.012% ethylenediaminetetraacetic acid produced an inhibitory effect on both the phages and the bacteria. PMID- 16349771 TI - Experimental evaluation of liquid film resistance in oxygen transport to microbial cells. AB - A membrane probe was used to monitor the dissolved oxygen concentrations in continuous cultures of Candida utilis and Micrococcus roseus growing at low dissolved oxygen concentrations and various agitation levels. For the yeast fermentations, increasing the agitation level within the range of 0.1 to 0.3 w per liter lowered steady-state dissolved oxygen concentrations in the fermentor. The steady-state dissolved oxygen concentration in the fermentor was not influenced by the agitation level within the range of 0.3 to 1.8 w per liter. With M. roseus, no effect of agitation on steady-state dissolved oxygen concentrations in the fermentor was observed within the range of 0.1 to 1.8 w per liter. It was concluded that, under the conditions used, a measurable transfer barrier from the liquid to the yeast cells existed at agitation levels below 0.3 w per liter and that this barrier did not exist at agitation levels above 0.3 w per liter. The transfer barrier from the liquid to the yeast surface could be represented by a stagnant film of liquid 0.6 x 10 cm thick surrounding the cell at an agitation level of 0.10 w per liter. This film represented an oxygen concentration drop of 1.3 x 10 M from the bulk of the medium to the cells under the experimental conditions. PMID- 16349772 TI - Isolation of Sphaerophorus necrophorus from Bovine Liver Abscesses. PMID- 16349773 TI - Biodegradation of Dichlorodiphenyltrichloroethane: Intermediates in Dichlorodiphenylacetic Acid Metabolism by Aerobacter aerogenes. PMID- 16349774 TI - Plaquing of viruses in the absence of a carbon dioxide atmosphere. PMID- 16349775 TI - Growth of Moth Cells in Suspension in Hemolymph-free Medium. PMID- 16349776 TI - Preparation of bacterial Fatty Acid methyl esters for rapid characterization by gas-liquid chromatography. PMID- 16349777 TI - Low-temperature incubation using a water supply. PMID- 16349778 TI - Chemical characterization of pooled animal sera. PMID- 16349779 TI - Pseudo-Precipitin Reactions of Rattlesnake Venom (Crotalus atrox) When Reacted with Certain Mammalian Sera. PMID- 16349780 TI - Plaque Formation with N-Tris(hydroxymethyl)methyl-2-aminoethanesulfonic Acid (Tes Buffer). PMID- 16349781 TI - Further Indications of Clostridium botulinum in Latin American Waters. PMID- 16349782 TI - Inducing L Forms in Listeria monocytogenes Types 1 through 7. PMID- 16349783 TI - Modification of the French pressure cell. PMID- 16349784 TI - Isolation of Pectinolytic Strains of Aerobacter aerogenes. PMID- 16349785 TI - Improved multipoint inoculating device for replica plating. PMID- 16349786 TI - Inhibition of Aflatoxin Synthesis by p-Aminobenzoic Acid, Potassium Sulfite, and Potassium Fluoride. PMID- 16349787 TI - Production of Malformin by Aspergillus awamori. PMID- 16349788 TI - Spleen-derived Rauscher Murine Leukemia Virus. PMID- 16349789 TI - Synthetic and complex media for the rapid detection of fluorescence of phytopathogenic pseudomonads: effect of the carbon source. PMID- 16349790 TI - Trace metal analysis of nutrient broth. PMID- 16349791 TI - Survival of poliovirus in flowing turbid seawater treated with ultraviolet light. AB - [This corrects the article on p. 533 in vol. 15.]. PMID- 16349792 TI - Identity of Culture Producing Antibiotic PA 1008 (PA 108). PMID- 16349793 TI - Effects of Culture Conditions on Production of trans-Cinnamic Acid from Alkylbenzenes by Soil Microorganisms. AB - The production of trans-cinnamic acid from various alkylbenzenes by soil microorganisms was studied intensively by use of a co-oxidation technique. The microorganisms were grown on n-paraffins, and they did not use aromatic compounds as a carbon source when the preferred substrate was present in the medium. The effects of cell population, co-oxidation time, and type and mode of addition of the alkylbenzenes on the yield of trans-cinnamic acid were investigated. Yields (5 g/liter) of a product consisting of trans-cinnamic acid (88 to 100%) and 5 phenylvaleric acid (0 to 12%) were obtained when the proper conditions were chosen. Of a variety of microorganisms studied, a soil isolate closely related to Cellulomonas galba was found to be best for the production of trans-cinnamic acid. PMID- 16349794 TI - Distribution of thiamine, biotin, and niacin in the sea. AB - The distribution of thiamine, biotin, and niacin was investigated in surface waters collected from 20 stations in southeast Alaska, by use of bioassay methods. A marine yeast, Cryptococcus albidus, was used for thiamine, and mutants of the marine bacterium Serratia marinorubra were used for biotin and niacin. Thiamine was found in 6 (38%) of the samples in amounts ranging to 200 ng/liter; biotin was found in 8 (38%) of the samples in amounts ranging to 3.1 ng/liter. Niacin was found to be completely absent in all of the samples assayed. The significance of the presence of thiamine and biotin in the productivity of the area is discussed. PMID- 16349795 TI - Microbial growth in a fuel-water system containing polyesterurethane foam. PMID- 16349797 TI - Bacteriological Survey of the Frozen Prepared Foods Industry: III. Potato Products. AB - [This corrects the article on p. 1327 in vol. 15.]. PMID- 16349796 TI - Indigenous flora from human saliva. PMID- 16349798 TI - Oxidation of Alkylbenzenes by a Strain of Micrococcus cerificans Growing on n Paraffins. PMID- 16349799 TI - Photodynamic inactivation of newcastle disease virus with acridine orange. AB - [This corrects the article on p. 1453 in vol. 15.]. PMID- 16349800 TI - Growth and survival of fuel isolates in hydrocarbon-fuel emulsions. AB - Two fuel utilizers, a strain of Pseudomonas aeruginosa and a Hormodendrum sp., and two fuel isolates which did not use fuel, Staphylococcus and Bacillus spp., were tested for ability to survive and grow in systems containing emulsified or nonemulsified forms of JP-4 jet fuel. Neither emulsion (Alamac no. 1 or Alamac no. 2) supported microbial growth without a water phase. Growth of P. aeruginosa in liquid systems containing either emulsion was not significantly different from growth in liquid systems with nonemulsified fuel. The Hormodendrum sp. grew well in a liquid medium containing nonemulsified JP-4, but when either of the emulsions served as carbon source no growth was observed. However, good growth was noted on spread plates with either emulsion. Viable cells of Bacillus sp. did not increase over a 4-day period, and Staphylococcus sp. did not survive in liquid systems containing JP-4 or emulsions. PMID- 16349801 TI - Fermentation rates of grape juice: v. Biotin content of juice and its effect on alcoholic fermentation rate. AB - Microbiological analysis showed that juices from white grapes had less biotin than juices from red grapes. The biotin content of the juices of some varieties was significantly different from that of other varieties. We did not note any regional effects on the biotin content of the juices. Biotin content of the Cabernet Sauvignon grapes increased significantly with maturity, whereas the biotin content of a white variety did not. The biotin content, with the total nitrogen, can be used to estimate indirectly the yeast growth potential and hence to predict the fermentation rate of the juice. About 84% of the rate variation can be accounted for by the calculated regression equations. PMID- 16349802 TI - Bacteriology of manganese nodules: III. Reduction of MnO(2) by two strains of nodule bacteria. AB - MnO(2) reduction by aerobic growing cultures of Bacillus 29 and coccus 32, isolated from ferromanganese nodules, was assessed for 7 days. A 1-day lag was observed before the onset of MnO(2) reduction by either culture. Addition of HgCl(2) to a final concentration of about 10 M caused a rapid cessation of MnO(2) reduction by the growing cultures. Neither culture reduced MnO(2) when grown under continued anaerobiosis from the start of an experiment. However, if conditions were made anaerobic after MnO(2) reduction was initiated, reduction continued at a rate only slightly lower than that under aerobic conditions. Resting-cell cultures reduced MnO(2) equally well aerobically and anaerobically, provided that ferricyanide was present to serve as electron carrier. These findings showed that oxygen is needed for culture adaptation to MnO(2) reduction, and that oxygen does not interfere with microbial MnO(2) reduction itself. Both cultures caused sharp drops in the pH of the medium during MnO(2) reduction: with coccus 32, during the entire incubation time; with Bacillus 29, for the first 3 days. The E(h) of the medium fluctuated with either culture and never fell below 469 mv with Bacillus 29 and below 394 mv with coccus 32. The rates of glucose consumption and Mn release by Bacillus 29 and coccus 32 were fairly constant, but the rates of lactate and pyruvate production were not. Although acid production undoubtedly helped in the reduction of pyrolusite (MnO(2)) by the bacteria, it did not appear to be important in the reduction of manganese oxide in ferromanganese nodules, as shown by the results with a nodule enrichment. PMID- 16349803 TI - Correlation of taxonomic criteria for a collection of marine bacteria. AB - Numerical taxonomy was done on 208 strains of marine bacteria. The collection was segregated into eight groups, seven of which contained Vibrio sp. Nucleic acid base ratio studies on a typical Vibrio sp. from each group and other genera were done. The phenotypically different Vibrio sp. had a narrow range of base ratios. The other genera had base ratios more similar to the base ratios reported for their genus than to each other as marine bacteria. The taxonomic groups are compared with generic classification and the strains' sources of isolations. PMID- 16349805 TI - Fungi Isolated from Damaged Flue-cured Tobacco. AB - Species of Aspergillus were the most prevalent fungi isolated from 51 samples of damaged flue-cured tobacco of the 1966 U.S. crop, comprising 57% of the total isolates. Other prevalent fungi were Penicillium (16%), Alternaria (8%), Cladosporium (4%), and Chaetomium (4%). Members of the Aspergillus glaucus group were isolated most frequently from samples with moisture contents ranging from 18 to 28%, whereas Alternaria, Cladosporium, and Penicillium were isolated consistently from samples containing 24 to 32% moisture. Aspergillus niger was prevalent in tobacco ranging in moisture content from 18 to 30%. PMID- 16349806 TI - Fungal susceptibility of polyurethanes. AB - One hundred laboratory-synthesized polyurethanes were tested by a mixed-culture petri dish method for susceptibility to fungus attack. Polyether polyurethanes were moderately to highly resistant to fungal attack, whereas all polyester polyurethanes tested were highly susceptible. The susceptibility of the polyethers was related to the number of adjacent methylene groups in the polymer chain. At least two such groups were required for appreciable attack to occur. The presence of side chains on the diol moiety of the polyurethane reduced susceptibility. PMID- 16349807 TI - Fungal metabolites as indicators of quality of tomato products. PMID- 16349808 TI - Bloater Formation by Gas-forming Lactic Acid Bacteria in Cucumber Fermentations. AB - The formation of "bloaters" (hollow stock) in cucumbers brined for salt-stock purposes at 5 to 10% salt has been associated with gaseous fermentation caused chiefly by yeasts. Recently, serious early bloater damage, not attributable to yeasts, has been observed in commercial-scale experiments on control of bloaters in overnight dill pickles brined in 50-gal barrels at 3.0 to 4.5% salt. Growth of fermentative species of yeasts was effectively controlled by the addition of 0.025, 0.05, and 0.1% sorbic acid or its sodium salt. In contrast to this, the fermenting brines showed extremely high populations of acid-forming bacteria, identified as Lactobacillus plantarum, L. brevis, and Pediococcus cerevisiae. The gas-forming species (i.e., L. brevis) constituted a high proportion of the total populations. Representative isolates from 36 barrels of overnight dill pickles were tested for their ability to produce bloaters in 1-quart jars of pasteurized cucumbers equilibrated at 4 to 5% salt, 0.25% lactic acid, and pH 4.0. Bloaters, identical with those made by yeast cultures, were produced in all jars inoculated with L. brevis. No bloaters were produced by L. plantarum and P. cerevisiae. These results suggest that the control of bloater damage in cucumber fermentations, particularly at low salt concentrations, may necessitate inhibition of gas-forming lactic acid bacteria. PMID- 16349809 TI - Hemagglutination and hemolysis by : lichen extracts. AB - Twenty-two species of lichens from 10 different genera possessed a hemagglutinin for one or more of human, sheep, horse, cow, rabbit, guinea pig, and chicken erythrocytes. Hemolysins were also detected occasionally, but these were only active at low dilutions. In those species tested, the hemolytic principle was dialyzable; the hemagglutinating agent was not. Preliminary studies have indicated that the lichen hemagglutinins are nonspecific. PMID- 16349810 TI - Direct quantitative gas chromatographic separation of c(2)-c(6) Fatty acids, methanol, and ethyl alcohol in aqueous microbial fermentation media. AB - [This corrects the article on p. 286 in vol. 16.]. PMID- 16349811 TI - Fungi isolated from tobacco leaves and brown-spot lesions before and after flue curing. AB - Discs of tissue excised from ripe nonflue-cured and flue-cured tobacco leaves, harvested on six dates and cultured on three selective media, yielded 21 and 24 genera of fungi, respectively. Of 5,094 fungi isolated from 3,240 pieces of nonfluecured leaf tissue, 89.5% comprised five genera, Alternaria, Cladosporium, Epicoccum, Trichoderma, and Nigrospora. Of 2,494 fungi isolated from 3,240 pieces of flue-cured leaf tissue, 70.9% were Alternaria, Cladosporium Epicoccum, Aspergillus, and Nigrospora. Flue-cured and nonflue-cured brown-spot lesions harvested at two locations yielded 12 and 14 genera of fungi, respectively. Alternaria, Penicillium, Phoma, and Stemphyllium comprised 91.5% of the 2,245 fungi isolated from noncured and 87.1% of the 1,118 fungi isolated from the cured lesions. The number and kinds of fungi obtained from diseased and healthy tissue were reduced but not eliminated by flue-curing. PMID- 16349812 TI - Regeneration of oxidative assimilation capacity by intracellular conversion of storage products to protein. AB - Previous studies have shown that the capacity of a heterogeneous microbial population for oxidative assimilation of glucose can be renewed by periodically subjecting the sludge (or a portion of it) to endogenous respiration in the presence of an exogenous source of nitrogen. Further study of this system led to a modification of the activated sludge process for nitrogen-deficient wastes. However, it was not known whether renewal of oxidative assimilation capacity was possible for substrates which required the presence of inducible enzyme(s) or for substrates which were not carbohydrates. Therefore, studies with lactose and acetate as carbon sources were designed. Both carbon sources were removed under conditions of oxidative assimilation, and the storage products (or a portion of these products) were converted into protein when the sludge was subjected to a period of endogenous respiration (with respect to carbon source) in the presence of an exogenous supply of ammonium sulfate. The "regenerated" sludge exhibited a renewed capacity for oxidative assimilation, thereby indicating that requisite inducible enzymes (e.g., beta-galactosidase in the case of lactose; iso-citritase and malate synthetase in the case of acetate) were not diluted out in the endogenous phase to a degree sufficient to hamper renewed oxidative assimilation capacity. The results also indicated that a noncarbohydrate carbon source can be successfully removed from the medium with this process. However, in the case of acetate, the oxidative assimilation capacity after "regeneration" was not fully restored to the initial level. PMID- 16349813 TI - Continuous oxidative assimilation of acetic Acid and endogenous protein synthesis applicable to treatment of nitrogen-deficient waste waters. AB - A previous study indicated that the oxidative assimilation capacity of a heterogeneous microbial population for acetic acid did not return to the initial value after a period of endogenous respiration in the presence of ammonium sulfate. In view of this finding it was not possible to recommend fully the use of a continuous oxidative assimilation process for the treatment of nitrogen deficient waste waters of noncarbohydrate nature. To put the process to a severe test, a laboratory-scale pilot plant study was done. Acetate (1,000 mg/liter) was fed continuously to a completely mixed, aerated reactor vessel, from which the mixed liquor was channeled to a settling basin. The settling basin supernatant fluid was continuously discharged, and a portion of the settled biological sludge was subjected to endogenous respiration in the presence of ammonium sulfate and was recycled to the aeration vessel. Experiments were conducted at three levels of ammonia nitrogen in the endogenous phase. Approximately 90% removal of the carbon source (expressed as chemical oxygen demand, COD) was attained with a low level of supplemental nitrogen (COD/N = 70:1) and a fairly low reactor detention time (4 hr). Based upon these and previous results, it has been concluded that the process shows promise for application to the treatment of a wide variety of nitrogen-deficient industrial wastes. PMID- 16349814 TI - Some properties of cellulolytic cellvibrio strains from polluted water. PMID- 16349815 TI - Simple circulating tank test for evaluation of germicides in cutting fluid emulsions. PMID- 16349816 TI - Occurrence of Bacterial Decay in Copper-Chrome-Arsenic-treated Wood. PMID- 16349817 TI - Motile Marine Bacteria: III. Effect of Storage of Seawater and Source of Isolation. PMID- 16349818 TI - Enumeration of Autotrophic Ammonium-oxidizing Bacteria in Marine Waters by a Direct Method. AB - A membrane filter method was developed for enumerating, in mixed populations, autotrophic nitrifying bacteria which transform ammonium to nitrite. It gave counts in good agreement with estimates obtained by using a most-probable-number procedure. Compared to the most-probable-number procedure, the membrane method is convenient and precise. A means of extending the sensitivity of the membrane method is presented. PMID- 16349819 TI - Energy utilization for polysaccharide synthesis by mixed rumen organisms fermenting soluble carbohydrates. AB - Synthesis of reserve polysaccharide by mixed rumen organisms fermenting glucose, maltose, cellobiose, and xylose has been studied in relation to the adenosine triphosphate energy calculated to be available from substrate fermentation. About 80% of the energy available from glucose and xylose was used for polysaccharide synthesis, whereas, assuming hydrolytic cleavage of the disaccharides, more than 100% was used when cellobiose and maltose were the substrates. If, however, phosphorolytic cleavage of the disaccharides, for which there is evidence, was involved, the energy from both maltose and cellobiose fermentation was used with about the same efficiency as that from glucose and xylose fermentation. The rumen fluid used was collected 24 hr after feeding, and growth of microorganisms in such samples was sufficient to account for utilization of less than 10% of the total energy becoming available during the 40-min incubation period. PMID- 16349820 TI - Starch Hydrolysis by Conidia of Aspergillus wentii. AB - Soluble starch was hydrolyzed to glucose by conidia of Aspergillus wentii NRRL 2001. Peak yields of glucose were achieved in 3 days. A glucoamylase-like enzyme was assumed to be responsible since maltose was not detected during the conversion. Spore age, storage conditions, and temperature affected the level of glucose accumulated. Iodoacetate inhibited catabolism of the glucose formed and this inhibition increased product yield. Spores of other fungi also hydrolyzed starch but none accumulated glucose naturally as did A. wentii spores. PMID- 16349821 TI - Murine Toxicity of Cochliobolus carbonum. AB - Seventeen wild-type strains of the phytopathogenic fungus Cochliobolus carbonum, tested by intraperitoneal injection into mice, were lethal within 48 hr. The lethal effect appeared to be a toxic rather than an infectious process, because death occurred within 3 hr after injection of two of the isolates and heat-killed cultures were lethal. Assays of ascospore progeny from two crosses involving three isolates indicated that the toxic metabolites were under genetic control and quantitative regulation. Studies of the toxicological, cultural, and chemical characteristics of these three strains indicated that more than one murine toxin was present. PMID- 16349822 TI - Penicillium mycelium waste as protein supplement in animals. AB - Dried Penicillium mycelium served as a protein source in animal diet when it was supplemented at 7.5% protein level along with 7.5% protein level from peanut meal. Under these conditions, the food consumption was optimal, and the rat growth response was comparable with 15% casein diet. The role of peanut meal appears to be twofold; it makes the mycelium diet more palatable and it supplies protein. The amino acids, lysine and threonine, which are found to be limiting in peanut meal, are reported to be present in the Penicillium mycelium. This type of formulation affords considerable economic advantage because both the peanut meal and the Penicillium mycelium are by-products and, therefore, are inexpensive sources of protein. PMID- 16349823 TI - Milk-clotting Enzyme from Microorganisms: V. Purification and Crystallization of Mucor Rennin from Mucor pusillus var. Lindt. AB - A rennin crystal was obtained from the crude milk-clotting enzyme of Mucor pusillus var. Lindt. The crude enzyme was purified by using columns of Amberlite CG-50, diethylaminoethyl Sephadex A-50, and Sephadex G-100. This purified enzyme was dissolved in 0.1 M sodium acetate (pH 5.0) buffer to a final concentration of 2 to 3%; ammonium sulfate (to 40% saturation) was added, and the resulting solution was placed in cellophane tubes. The enzyme solution was dialyzed against 0.1 M sodium acetate buffer (pH 5) containing ammonium sulfate was added dropwise to the outside solution of the cellophane tube, and the concentration of ammonium sulfate in the cellophane tube increased gradually. The crystals of enzyme were formed in the cellophane tube when the concentration reached approximately 50% saturation. After the enzyme solution was concentrated in the freezer, the crystals were obtained. The activity of the crystalline enzyme was inhibited by Hg, Ag, Zn, and KMnO(4). PMID- 16349824 TI - Use of chemical dispersants in the enumeration of bacteria in activated sludge. PMID- 16349825 TI - Increased recovery of psychrophilic bacteria by the use of a new medium with lower solidifying temperature. PMID- 16349826 TI - Effects of jet-fuel microbial isolates on a polyurethane foam. AB - Jet-fuel microbial isolates were studied for effects on a polyurethane foam material that has been proposed as a baffling material for use in aircraft fuel tanks. Evidence was found that a polyesterurethane foam gave increased cell counts and oxygen uptake with a bacterial isolate, and extensive matting with fragmentation and decreases in tensile strength of the foam with a fungal isolate. The polyurethane foam was affected by activity of the jet-fuel microbial isolates to an extent that would cause serious microbiological problems in the fuel tanks of jet aircraft. PMID- 16349827 TI - Water relations of xerophilic fungi isolated from prunes. AB - The predominant spoilage fungi of dried and high-moisture prunes were members of the Aspergillus glaucus group and Xeromyces bisporus. Chrysosporium spp. were also important. At the mean pH of prune flesh (3.8) and at 25 C, X. bisporus grew at water activities (a(w)) down to 0.605, and Chrysosporium fastidium grew to 0.686. Germination was always followed by growth, but within the 120-day incubation period, the minimum a(w) permitting asexual sporulation was usually higher than that permitting germination. Sexual sporulation often required an even higher a(w). The water requirements of aspergilli were appreciably greater at this pH than near neutrality, no species germinating below 0.738 a(w). This was probably a consequence of a high spore-death rate during incubation at low a(w) and pH. PMID- 16349828 TI - Characteristics of myxobacteria isolated from the surface of freshwater fish. AB - A study was made of 32 nonpathogenic myxobacterial isolates obtained from a variety of fish taken in the Pacific Northwest. Morphological, cultural, biochemical, and serological studies were carried out on these strains. All were found to be members of the genus Cytophaga. Two myxobacterial strains pathogenic to fish were also included in this study for comparative purposes. These pathogenic organisms were found to be culturally and physiologically similar to some of the nonpathogenic strains. Antiserum against the pathogenic species, however, showed no cross agglutination when tested against the other myxobacterial isolates. As a result, serological procedures appear promising as a rapid means for distinguishing pathogenic myxobacteria from one another and from saprophytic myxobacteria commonly found on fish. PMID- 16349829 TI - Effect of Methionine and Sulfate on the Metabolism of Cephalosporium acremonium. AB - The metabolism of Cephalosporium acremonium grown in a complex medium supplemented with DL-methionine or inorganic sulfate was studied. More growth occurred in a sulfate medium than in a methionine medium. Methionine-grown cells had an increased rate of respiration, a higher rate of catabolism with acetate and glucose as substrate, and higher specific activities of certain respiratory enzymes than sulfate-grown cells. Labeled acetate and glucose were assimilated at a faster rate by methionine-grown cells than sulfate-grown cells. Taurine, cystathionine, and small quantities of four acidic compounds were present in the amino acid pool of methionine-grown cells, but they were not detected in the pool of sulfate-grown cells. The differences in metabolic activity of sulfate and methionine-grown cells are discussed in regard to cephalosporin C synthesis. PMID- 16349830 TI - Effects of storage of linen straws on retting process. PMID- 16349831 TI - Fermentation of philippine vegetable blends. AB - Seven blends of Philipphine vegetables, two of which contained soybeans and one mongo bean sprouts, were prepared for fermentation and study of microbiological and chemical changes. The fermentations were typical lactic acid bacterial fermentations, initiated by Leuconostoc mesenteroides and continued by Lactobacillus brevis, Pediococcus cerevisiae, and L. plantarum. The combination of high acidity and low pH resembled other vegetable fermentations, such as sauerkraut. The procedure offers a method of preserving surplus vegetables, and, in addition, a method for incorporating and preserving the high-protein containing soybeans. PMID- 16349832 TI - Effect of soya flour on the lactic fermentation of milled corn. PMID- 16349833 TI - Production of Nuclease-forming 5'-Nucleotide by Aspergillus quercinus in a Low Phosphate Medium. AB - [This corrects the article on p. 1293 in vol. 16.]. PMID- 16349834 TI - Antifungal properties of cranberry juice. AB - [This corrects the article on p. 1524 in vol. 16.]. PMID- 16349835 TI - Retting of Flax by Aspergillus niger. AB - In this study, retting was carried out by Aspergillus niger. The pH, galacturonic acid (GA), and total reducing sugar were determined; the end point was identified by the classic empirical processes and by the maximal GA content of the retting water. The process gave clear and resistent fibers, and the retting time was similar to that of current industrial processes with bacterial enzymes. Control of total acidity was not required, since the pH remained close to neutrality throughout the entire process. PMID- 16349836 TI - Effects of polyurethane foams on microbial growth in fuel-water systems. AB - Four, open-cell, ester-base polyurethane foams were examined for their effect on growth of fuel-utilizing organisms in jet fuel-water systems. Three foams contained a potential biocide, tetraethylthiuram E (0.66%), sodium omadine (0.07%), or zinc omadine (0.07%), all w/v. These were compared with a control foam which did not contain an additive. Each foam was examined in fuel-water systems containing JP-4 fuel, JP-4 fuel plus 0.1% anti-icing additive (AIA), or JP-5 fuel. Pure cultures of a fuel-grown bacterium, Pseudomonas aeruginosa, and of a fuel-grown fungus, Hormodendrum (Cladosporium) sp., served as test organisms. In control cultures without foam and in cultures containing control foam, P. aeruginosa achieved maximum stationary-phase populations of approximately 10 viable cells per ml, and Hormodendrum sp. produced an extensive mycelial mat. In the three fuel systems examined, tetraethylthiuram E- and sodium omadine-containing foams had little effect on growth of the bacterium; foam with zinc omadine decreased the rate of bacterial growth but had little effect on total populations. Tetraethylthiuram E decreased the rate of fungal growth and showed its greatest effect in JP-4 plus AIA. Foam with sodium omadine or zinc omadine markedly decreased fungal growth in all three fuel systems. The data suggest that either sodium omadine or zinc omadine in polyurethane foam may be a useful antifungal agent; and that tetraethylthiuram E and AIA could exert a synergistic effect, particularly at AIA concentrations which have been reported to occur in some field situations. PMID- 16349837 TI - Marine microorganisms associated with the food of young salmon. AB - The biomass of microorganisms in the digestive tracts of young salmon in the sea of Saanich Inlet was examined by (i) the direct microscopic method, (ii) the plate count method and (iii) the physiological method (microbial biomass assumed from the release of carbon dioxide). By all methods, most microbial biomass in foods was shown to decrease during its stay in the digestive organ. This observation was supported by the detection of many bacteria at different stages of digestion. The microbial biomass was quantitatively influenced by the composition of food ingested and by the sampling time. PMID- 16349838 TI - Identification of Nonfermenting Gram-negative Bacilli in the Clinical Laboratory. PMID- 16349839 TI - Improving reliability in penicillin assays. AB - [This corrects the article on p. 1941 in vol. 16.]. PMID- 16349840 TI - Standardized, automated procedure for measurement of serum hemolytic complement activity. AB - [This corrects the article on p. 1635 in vol. 16.]. PMID- 16349841 TI - Fungi Isolated from Flue-cured Tobacco at Time of Sale and After Storage. AB - The fungi isolated from 100 samples of flue-cured tobacco from 12 markets in 2 tobacco belts comprised 11 genera, including 10 species of Aspergillus. The mean percentage per sample isolated from 62 samples of tobacco from Middle Belt markets was Alternaria, 40.6%; Aspergillus niger, 47.8%; Aspergillus repens, 38.0%; and Penicillium, 25.8%. The mean percentage per sample isolated from 38 samples of tobacco from Old Belt markets was Alternaria, 74.0%; Penicillium, 52.5%; Aspergillus repens, 38.0%; and Aspergillus ruber, 36.2%. Damaged (74 samples) and nondamaged (26 samples) stored tobacco yielded species of six genera of fungi, including eight species of Aspergillus. Species of Aspergillus and Penicillium were commonly isolated from both damaged and nondamaged tobacco, whereas species of Alternaria, Cladosporium, Fusarium, and Rhizopus were isoalted more frequently from nondamaged tobacco. The fungi that occurred in the highest population in damaged tobacco were Aspergillus repens, A. niger, A. ruber, and Penicillium species. PMID- 16349842 TI - Identification of pseudomonas species isolated from hospital environment and human sources. AB - [This corrects the article on p. 1535 in vol. 16.]. PMID- 16349844 TI - Oxygen-tolerant Strain of Chlorella sorokiniana. AB - [This corrects the article on p. 140 in vol. 17.]. PMID- 16349843 TI - Mode of action of the antibacterial compound dequalinium acetate. AB - [This corrects the article on p. 122 in vol. 17.]. PMID- 16349845 TI - Influence of pasteurization and lye treatment on the fermentation of spanish style manzanilla olives. AB - The effect of lye concentration and pasteurization on the fermentation of Spanish style Manzanilla green olives was investigated. Results showed that a rapid, normal fermentation always occurred when the proper lye was used, and only in the case of an inappropriate alkaline treatment could pasteurization help the growth of lactobacilli. Although pasteurization does not seem to affect color of processed olives, texture tests gave lower values for pasteurized samples than for those obtained from nonpasteurized olives. PMID- 16349846 TI - Biological response of lactic streptococci and lactobacilli to catalase. AB - Addition of catalase to milk cultures of lactic streptococci resulted in increased rates of acid production, although it had no effect on cultures of lactobacilli. Milk cultures of both streptococci and lactobacilli produced detectable amounts of peroxide, which reached a maximum level in the early period of acid production followed by a drastic decrease as the acid production increased. Pyruvate and reduced glutathione decreased the amount of peroxide formed, but had little effect on acid production by the streptococci. Ferrous sulfate prevented the accumulation of peroxide and stimulated the rate of acid production by the streptococci to a greater extent than did catalase. PMID- 16349847 TI - Preparation of concentrated lactic streptococcus starters. AB - Single-strain cultures of Streptococcus cremoris were grown in a semisynthetic medium with automatic pH control. After centrifugation, the cells were resuspended in sterile nonfat milk (2% of the original volume). There was no significant difference in the maximum population attained when cultures were grown at pH values of 5.5, 6.0, or 6.5 with sodium hydroxide as the neutralizer. With ammonium hydroxide as the neutralizer, maximum populations obtained were increased about twofold. In most cases, the acid-producing ability of the culture concentrates was comparable to that of fresh-milk cultures. There was some variation among strains of S. cremoris with respect to the effects of different neutralizers and levels of pH control on the biological activity of the culture concentrates. The culture concentrates were stored in liquid nitrogen for as long as 231 days without significant loss in biological activity. PMID- 16349848 TI - Growth and Pigmentation of Epicoccum nigrum in Submerged Culture. AB - Growth and pigmentation of Epicoccum nigrum was studied in submerged culture in various media. Red pigmentation of the mycelium and of fermentation broth was obtained only in a medium containing glucose and yeast autolysate. This pigmentation occurred at the time of maximal production of carotenoids. Observations on the submerged culture of this organism in flasks and in fermentors are described, and the details of a standardized procedure for the production of carotenoid pigments are given. PMID- 16349849 TI - Microbial Hydrocarbon Co-oxidation. III. Isolation and Characterization of an alpha, alpha'-Dimethyl-cis, cis-Muconic Acid-producing Strain of Nocardia corallina. AB - A soil isolate identified as a strain of Nocardia corallina accumulated alpha, alpha'-dimethyl-cis, cis-muconic acid under co-oxidation conditions employing n hexadecane for growth and p-xylene as the co-oxidizable substrate. N. corallina V 49 was postulated to have two pathways for the oxidation of p-xylene. One pathway proceeds throughp-benzyl alcohol, p-tolualdehyde, and p-toluic acid to 2, 3 dihydroxy-p-toluic acid, and the other pathway results in ortho ring cleavage of 3, 6-dimethylpyrocatechol and hence accumulation of alpha,alpha'-dimethyl-cis, cis-muconic acid. PMID- 16349850 TI - Production and Properties of alpha-L-Arabinofuranosidase from Corticium rolfsii. AB - When Corticium rolfsii is grown under aerobic conditions in a medium containing one of several simple sugars or polysaccharides, it release alpha-L arabinofuranosidase into the culture fluid. Araban and bran extract were found to be the most effective carbon sources in stimulating the production of the enzyme. Pectin and arabinose stimulated the production of the enzyme to a lesser degree, whereas xylose, glucose, galactose, and sucrose caused the formation of a relatively small amount of alpha-L-arabinofuranosidase. alpha-L Arabinofuranosidase was demonstrated by its ability to hydrolyze phenyl-alpha-L arabinofuranoside, araban, and arabinoxylan. The pH optimum of the enzyme was 2.5. At pH values of 2 to 9, the enzyme lost less than 15% of its activity during a 72-hr period at 2 C. At 70 C, its stability was greatest at pH values of 4 to 6. PMID- 16349851 TI - Lipase from Pseudomonas fragi. I. Purification of the Enzyme. AB - The experimental conditions required to isolate a lipase from Pseudomonas fragi were determined. The organism was grown in a buffered tryptone medium for 4 to 5 days at 20 C. The lipase in the culture supernatant fluid was isolated by fractionation with ammonium sulfate at 60% saturation, followed by acetone precipitation at 30-60% concentration. Further purification was made by using Sephadex G-200 gel-filtration and diethylaminoethyl cellulose chromatography. Electrophoretic analysis of the purified lipolytic fraction showed apparent homogeneity by both cellulose polyacetate and disc electrophoresis. The specific activity of the purified enzyme was about 100 times that of the starting culture filtrate, and the yield was about 1.8% of the original activity. PMID- 16349852 TI - Lipase from Pseudomonas fragi. II. Properties of the Enzyme. AB - The optimal pH value of a lipase from Pseudomonas fragi was between 7.5 and 8.9, and a high reaction rate was observed at 54 C. Heating the enzyme solution at 63 C for 30 min inactivated only 27.6% of its activity; however, total inactivation was observed at 66 C after 1 hr and at 71 C after 10 min. The lipase was inhibited strongly by Fe and Fe ions, and to a lesser extent by Co, Cu, Zn. No inhibition was observed with Ca or NaF. Ethylenediaminetetraacetate was effective in removing the toxicity of Fe. The activity of the enzyme was inhibited markedly by p-chloromercurobenzoate, but the effects of N-ethylmaleimide and iodoacetate were moderate. The enzyme was able to hydrolyze natural fats, synthetic triglycerides, and alcohol esters. The order of the rate of hydrolysis of some triglycerides under experimental conditions was, from the fastest to the lowest, trilaurin, tricaprin, tricaprylin, tripalmitin, tributyrin, tricaproin, and tristearin. The enzyme was capable of hydrolyzing methyl butyrate, but the rate of hydrolysis was about one-fifth that for triolein and one-thirteenth that for coconut oil. The enzyme lost its activity rapidly when held frozen, at 20 C, and at the extremes in pH. Glutathione, cysteine, and mercaptoethanol did not preserve the activity of the enzyme. PMID- 16349853 TI - Effect of pH on the Preservation of Palm Wine by Sulfite. AB - A study of the effect of pH on the preservation of palm wine by sulfite was made. It was concluded that at pH 4.0 to 5.0, when the palm wine is considered most suitable for drinking, the concentration of the sulfite required to suppress microfloral activities would be excessive for human consumption. PMID- 16349854 TI - Immunofluorescence Identification of Thermopolyspora polyspora, the Causative Agent of Farmer's Lung. AB - [This corrects the article on p. 454 in vol. 17.]. PMID- 16349855 TI - Enzymatic reduction of iron oxide by fungi. AB - The occurrence of the iron-reducing phenomenon among some common fungi was studied. Results indicated that (i) the reduction of ferric iron to the ferrous state by fungi seems to be restricted to nitrate reductase-inducible strains such as Actinomucor repens, Alternaria tenuis, Fusarium oxysporum, and F. solani and (ii) the amount of dissolved ferrous iron may be reduced progressively by increasing the amount of nitrate added to the medium. Compared with a complex medium (Sabouraud medium), less iron became reduced if NO(3) was the only nitrogen source (Czapek Dox medium). These data strongly support the view that ferric iron is acting as an hydrogen acceptor in respiration, competing with nitrate for electrons that are mediated by the enzyme nitrate reductase. The significance of this property from an ecological viewpoint is discussed. PMID- 16349856 TI - Control of Byssochlamys and Related Heat-resistant Fungi in Grape Products. AB - Heat-resistant strains of Byssochlamys fulva, B. nivea, and other heat-resistant fungi were isolated from vineyard soil, grapes, grape-processing lines, and waste pomace. They are known to remain in grape juice occasionally and to grow in grape juice products. Ascospores of these fungi have a D value (decimal reduction time) of about 10 min at 190 F (88 C), but in the presence of 90 muliters of SO(2) per liter (normally added to the juice) the D value was cut in half. Filtration through a commercial diatomaceous filter aid (also a common processing step) entrapped all but about 0.001% of experimentally added spores. Thus, heat in the presence of SO(2) and filtration together can reduce the population of these spores by several orders of magnitude. Growth was also prevented by benzoate or sorbate in low concentrations. Oxygen must be reduced to extremely low levels before lack of oxygen limits growth. PMID- 16349857 TI - Effects of nitrogen limitation on the growth and composition of unicellular algae in continuous culture. AB - Since fat accumulation takes place in many algae as a response to exhaustion of the nitrogen supply, it has been suggested that this may provide a means of enhancing the potential food value of algae. To test this possibility, chemostatic continuous cultures of Chlorella sorokiniana and Oocystis polymorpha were subjected to successive reductions in influent nitrogen. As cellular nitrogen content decreased from about 10 to 4%, oxygen evolution, carbon dioxide uptake, chlorophyll content, and tissue production were drastically reduced, but total lipid content was essentially unchanged. Caloric values and C, H, and N analyses suggested a moderate increase in carbohydrate content, but gas chromatographic analyses revealed no significant qualitative or quantitative changes in the fatty acid fraction. In batch-cultured cells, nitrogen could be reduced to 3% of dry weight, causing a concomitant increase in total fatty acids and pronounced changes in the composition of the fatty acid fraction. These results suggest that cellular nitrogen must fall to approximately 3% of dry weight before appreciable increases in lipid synthesis can occur. Cell nitrogen is then apparently completely bound in essential cell constituents, and carbon subsequently fixed is converted into lipid products. The findings indicate that nitrogen limitation may be useful in increasing the food quality of batch cultured cells, but the technique has little value for continuous culture systems per se. PMID- 16349858 TI - Immunological studies with an m-deficient histoplasmin skin-test antigen. AB - Previous studies have shown that a single skin test to histoplasmin may induce complement-fixing antibodies or M precipitins (or both) to histoplasmin in histoplasmin-sensitive, but serologically negative, individuals. Ideally a skin test antigen should be one which detects hypersensitivity without stimulating humoral antibodies. Histoplasmin skin-test antigens presently used contain both H and M antigens. The present study was undertaken to evaluate an histoplasmin skin test antigen deficient in the M component but containing the H antigen. Thirty histoplasmin-hypersensitive subjects were bled prior to administration of the experimental skin-test antigen and at various time intervals thereafter. Only six of the thirty hypersensitive subjects showed serological responses. The sera of the six, however, only showed weak precipitin reactions, five showed M bands, and only one showed an M and an H band. None showed complement-fixation titers with either the yeast or mycelial antigens of Histoplasma capsulatum. Our data suggest that the use of a skin-test antigen purified to contain only H component would detect histoplasmin hypersensitivity without inducing antibodies and would eliminate false-positive serological reactions caused by the M component. PMID- 16349859 TI - Fermentation patterns of poorly fermenting cabbage hybrids. AB - Thirteen varieties of cabbage, including five commonly used commercial varieties and eight newly developed hybrids, were analyzed for their ability to undergo lactic acid fermentations. The rates of fermentation, total quantities of acids produced, and chemical analyses indicate that 11 of the 13 selections could support adequate fermentation at any of the three stages of maturation. However, two varieties G27 x G51 (105) and G60 x W-1 (106), harvested during the earlier phases of the 1967 and 1968 growing seasons, failed to undergo proper fermentations and consistently resulted in producing putrid products. These hybrids when harvested at the late stage of maturation were capable of producing total titratable acidities comparable to the acceptable varieties. However the resulting sauerkrauts were judged to be of poor quality. The inability of these varieties to support adequate fermentations may be due to growth inhibitory substances or the lack of inherent maturation-dependent growth factor(s) essential for the growth of lactic acid bacteria. PMID- 16349860 TI - Predominant Catalase-negative Soil Bacteria. III. Agromyces, gen. n., Microorganisms Intermediary to Actinomyces and Nocardia. AB - The occurrence of filamentous, branching, catalase-negative bacteria as a numerically predominant microflora of various soils was demonstrated by using a dilution frequency isolation procedure. The major characteristics of these organisms were those of the order Actinomycetales. However, they could not be placed in any of the present genera of this order and, therefore, a new genus, Agromyces, was proposed for these organisms. This genus includes catalase negative, nutritionally-fastidious microorganisms whose cells produce a true branching mycelium that fragments into coccoid and diphtheroid forms. Also, they have an oxidative metabolism, are microaerophilic to aerobic, and contain neither diaminopimelic acid nor lysine as major constituents of the cell wall glycopeptide. The type species would be Agromyces ramosus, gen. n., sp. n. The possible importance of these organisms in clarifying certain phylogenetic relationships of the Actinomycetales is discussed. PMID- 16349861 TI - Nitrate ion-selective electrode in microbial media. AB - A nitrate ion-selective electrode has been used to follow the kinetics of the reduction of nitrate by a bacillus isolated from the rumen of a sheep. The electrode system and the microbial culture were found to be quite compatible, although the formation of nitrite limits the utility and precision of the measurement. The formation of nitrite was followed simultaneously by a colorimetric method. Nitrate was converted quantitatively to nitrite. The rate of the reaction was found to increase to a plateau value, then drop abruptly when the nitrate was exhausted. PMID- 16349863 TI - Use of a rapid fluorescent-antibody staining technique with group a streptococci. AB - [This corrects the article on p. 940 in vol. 17.]. PMID- 16349862 TI - Production of m-Cresol by Valsa friesii. AB - m-Cresol has been isolated from culture filtrates of Valsa friesii. The yield after 10 days of growth on glucose-phytone broth was 160 mg/liter. PMID- 16349864 TI - Some pink yeasts associated with softening of olives. AB - Pink yeasts identified as Rhodotorula glutinis var. glutinis, R. minuta var. minuta, and R. rubra produce polygalacturonases which cause a slow softening of olive tissue. Both pectin methyl esterase and polygalacturonase are produced when cultures are grown in appropriate media. Crude, cell-free dialyzed enzyme preparations measured viscosimetrically exhibited optimal activity on sodium polygalacturonate at pH 6.0 and 40 C, and were active in the range of pH 4.0 to 9.0 and 10 to 50 C. Cultures grown in sterilized olives and brine at pH 4.0 with sterile glucose added aseptically caused a slow softening of tissue as measured with a Christel texturometer. Similar results were obtained when crude, cell-free enzyme preparations were added to olives in buffer solution at pH 6.0 with Merthiolate. Commercial control of these yeasts is easy if anaerobic conditions can be provided. Otherwise, the industry has to resort to manual removal of the film from the brine surface, either by skimming or by flagellation. PMID- 16349865 TI - Control mechanisms operative in a natural microbial population selected for its ability to degrade L-lysine. I. Effect of glucose in batch systems. AB - A natural microbial population was selected in a medium containing L-lysine as the sole carbon source and ammonia as a nitrogen source. Cells were harvested from a batch-operated fermentor containing lysine and were grown through one transfer on lysine, glucose, or a mixture of lysine and glucose. By comparing the substrate removal rates and enzymatic capabilities of the cells, it was determined that the inducible enzyme system responsible for lysine degradation was subject to catabolic repression. Inhibition of the activity of preformed enzyme(s) played only a minor role. Preinduction by lysine offered only a small degree of protection against repression. The removal of ammonia nitrogen from the system did not overcome the effect of glucose. PMID- 16349866 TI - Control Mechanisms Operative in a Natural Microbial Population Selected for Its Ability to Degrade L-Lysine. II. Effects of Fructose and Ribose in Batch Systems. AB - A natural microbial population was acclimated to L-lysine as the sole carbon source when ammonia nitrogen was provided in the medium. Fructose exerted a slight retarding effect upon the metabolic removal of lysine. The response was due to catabolite repression of the inducible enzyme system responsible for lysine degradation. Inhibition of activity of preformed enzymes played no part in the response. Ribose caused a slight increase in the rate of synthesis of lysine degrading enzymes. PMID- 16349867 TI - Control Mechanisms Operative in a Natural Microbial Population Selected for Its Ability to Degrade L-Lysine. III. Effects of Carbohydrates in Continuous-Flow Systems Under Shock Load Conditions. AB - Two naturally selected microbial populations were maintained under continuousflow conditions with glucose or magnesium growth-limiting. The reactors were subjected to shock loads by changing the influent substrate from L-lysine to a mixture of L lysine and glucose, L-lysine and fructose, or L-lysine and ribose. During the subsequent transient state, the following parameters were examined: lysine chemical oxygen demand (COD), carbohydrate COD, total COD, biological solids concentration, cell protein, enzymatic capability (lysine-degrading enzymes), and the rate of lysine removal. The carbohydrate was then removed from the influent and the same parameters were examined until a new steady state was established. In all cases, glucose and fructose caused a significant repression of the synthesis of lysine-degrading enzymes, resulting in a decrease in the enzymatic capability of the cells. In the carbon-limited reactor, the faster the flow rate, the greater was the repression, whereas, in the magnesium-limited reactor, the slower the flow rate, the greater was the repression. The introduction of ribose into the reactors caused an initial increase in lysine enzymatic capability followed by a slight repression when ribose degradation started. PMID- 16349868 TI - Isolation of a bacterial inhibitor from green olives. AB - A compound inhibitory to lactic acid bacteria was isolated from green Manzanillo olives. The inhibitor is a phenolic compound, is devoid of acid-hydrolyzable reducing sugar, and has a bitter taste. Freezing the olives prior to extraction caused chemical changes which greatly increased the level of the inhibitor, whereas heating prior to freezing prevented its formation. PMID- 16349869 TI - Bacterial immunoabsorbent columns. AB - Methods are described for the preparation of bacterial immunoabsorbents consisting of organisms dispersed in agar beads 50 to 200 mum in diameter. The concentration of organisms could be made extremely high and columns prepared had sufficient capacity to enable them to be used for a number of purposes, but particularly for the removal of cross-reacting agglutinins from production batches of type-specific antisera. The absorptive capacity of these columns was regenerated after absorption by the elution of agglutinins with low pH buffers. Results obtained from the use of these immunoabsorbents are presented. PMID- 16349870 TI - Survival of Salmonella montevideo on Wheat Stored at Constant Relative Humidity. AB - Eleven samples of Ottawa variety hard red winter wheat were inoculated with a standardized suspension of Salmonella montevideo. The contaminated wheat samples were placed in constant relative humidity (RH) chambers held at 25 C. Relative humidities were 7, 11, 22, 33, 43, 53, 62, 75, 84, 92, and 98%. Constant RH at 25 C was maintained with different saturated salt solutions in the sealed chambers. Periodic counts of viable S. montevideo cells per gram of wheat were made over a 28-week sampling period. Viable counts of S. montevideo on wheat held at 7, 11, and 22% RH decreased from an initial 10 cells/g of wheat to a final count of 10 cells/g in each sample. Samples stored at 33, 43, 53, and 62% RH decreased from 10 viable cells/g to 3.6 x 10, 10, 10, and 20 viable cells/g, respectively. No viable S. montevideo cells were detected in the samples held at 75, 84, 92, and 98% RH after 22, 16, 26, and 16 weeks, respectively. PMID- 16349871 TI - Production of Aflatoxins by Aspergillus flavus Cultured on Flue-Cured Tobacco. AB - Under favorable growth conditions, Aspergillus flavus can produce aflatoxins on flue-cured tobacco leaves. PMID- 16349872 TI - Numerical Taxonomy of Heterotrophic Bacteria Growing in Association with Continuous-Culture Chlorella sorokiniana. AB - Twenty-nine cultures isolated from nonaxenic growths of Chlorella sorokiniana (Shihira and Krauss) and eight reference cultures were tested for 150 morphological and biochemical characteristics. The taxonomic data were subjected to computer analysis from which five major clusters were identified. The bacterial isolates have been placed in the genera: Pseudomonas, Acinetobacter, Flavobacterium, and Bacillus. Amino acid requirements of four strains from the test set were determined. PMID- 16349873 TI - Utilization of Methylthio-s-Triazine for Growth of Soil Fungi. AB - Aspergillus niger van Tieghem, Aspergillus tamarii Kita, and Aspergillus flavus Link ex Fries utilized the methylthio moiety of 2,4-bis(isopropylamino) -6-methyl mercapto-s-triazine (prometryne) as a sulfur nutrient source. Other soil fungal isolates not affected by prometryne concentrations to 1 mg/ml culture included: Aspergillus oryzae (Ahlburg) Cohn, Curvularia lunata (Wakker) Boedijn, Trichoderma viride Persoon ex Fries, Alternaria tenuis Nees ex Corda, Penicillium funiculosum Thom, and Paecilomyces varioti Bainier. PMID- 16349874 TI - Nutrition Requirements of Pleurotus flabellatus. AB - The mycelium of Pleurotus flabellatus was grown in a synthetic medium to obtain accurate information on its nutritional requirements. Among various carbon sources tried, the organism was found to utilize hexose sugars more readily than other sugars. Ammonium citrate was found to be the best source of nitrogen. The yield of dry matter increased as the concentration of nitrogen was increased up to a certain stage beyond which there was no increase in the yield, but the crude protein content of the mycelium increased. Detailed studies on the effect of varying the concentrations of other major nutrients, i.e., potassium, phosphorus, calcium, and magnesium, on the growth and crude protein content of the mycelium were also carried out. Optimal pH range was fairly broad, lying between 4.5 to 7.5. PMID- 16349875 TI - Effects of Hypobaric and Hyperbaric Helium Atmospheres on the Growth of Chlorella sorokiniana. AB - Growth of Chlorella sorokiniana is unaffected by the choice of atmospheric diluent gas. Reduced total pressure has no inhibitory effect and may be slightly stimulatory. PMID- 16349876 TI - Improved methodology for isolating soil microorganisms. AB - Polyurethane foam cylinders and a replica-stamp technique were used for plating soil samples. This in conjunction with the addition of NaCl to the isolation medium substantially increased the variety of isolates recovered. PMID- 16349878 TI - Analysis of the diluents used for rubella virus hemagglutination and hemagglutination-inhibition tests. AB - [This corrects the article on p. 222 in vol. 18.]. PMID- 16349877 TI - Lid method for evaluating zone of inhibition of treated fabrics. AB - Stainless-steel wire cloth lids were found to be advantageous in the evaluation of zones of inhibition with germicide-treated cotton fabrics. PMID- 16349879 TI - Measurement of CO(2) Assimilation in Soils: an Experiment for the Biological Exploration of Mars. AB - A method is described for the measurement of CO(2) assimilation by microorganisms in soils. A determination involves exposing soil to CO(2), pyrolyzing the exposed soil, trapping the organic pyrolysis products on a column of firebrick coated with CuO, combusting the trapped organics by heating, and measuring the radioactivity in the CO(2) produced in the combustion. The detection of significant levels of C in the trapped organic fraction appears to be an unambiguous indication of biological activity. The CO(2) which is adsorbed or exchanged into soils by nonbiological processes does not interfere. The method easily detects the CO(2) fixed by 10 to 10 algae after light exposure for 3 to 24 hr. Assimilation of C is also demonstrable in dark-exposed soils containing 10 to 10 heterotrophic bacteria. Possible applications of the method in the biological exploration of Mars are discussed. PMID- 16349881 TI - Bordetella pertussis Serotypes in the United States. AB - [This corrects the article on p. 620 in vol. 18.]. PMID- 16349880 TI - Effect of light intensity and thickness of culture solution on oxygen production by algae. AB - Data from a small cylindrical culture unit with variable annular culture chambers indicate that (i) the rate of oxygen evolution by an algal culture in the linear phase of growth is a logarithmic function of light intensity, and (ii) the rate of oxygen evolution per unit volume of suspension is linearly related to the reciprocal of culture thickness. These two relationships have been combined in an empirical equation which gives the expected variation of the oxygen production rate with light intensity, culture thickness, and suspension volume. The applicability of this equation has been tested on a larger, multilight culture unit in this laboratory. The agreement between the experimental and calculated oxygen production rates was very satisfactory, suggesting that the equation is not limited to a particular culture unit but may have wide applicability. The efficiency of the culture unit from the standpoint of oxygen output (chemical energy) relative to electrical energy to supply the light source has been calculated, and the maximum value of 0.51% was obtained. The energy to run auxiliary equipment was not a factor in these calculations. The maximum efficiency in converting light energy to chemical energy was approximately 12%. An extrapolation of the experimental results suggests that approximately 2 ft and 30 kw would be required to provide the oxygen needs of one man. PMID- 16349882 TI - Microbial biomass on particulate organic matter in seawater of the euphotic zone. AB - Microbial biomass on suspended organic matter in seawater of the euphotic zone of Saanich Inlet was investigated. The viable microorganisms were measured by the glucose-uptake method. Microbial carbon on particulate organic matter in seawater was determined to be, on the average, 9.9 mug of C/liter, and there was a regression relationship as y = 0.0062 x - 1.79 with an unbiased variance V(yx) = 0.38, where x = particulate organic carbon in seawater (micrograms of C/liter) and y = logarithm of microbial carbon (micrograms of C/liter). PMID- 16349883 TI - Resistance of weathered cotton cellulose to cellulase action. AB - Increased resistance of weathered cotton cellulose to microbial breakdown has been shown to be the result of development of resistance to the action of fungal cellulases. Photochemical activity during weathering exposure transforms the cellulose into an altered substrate that prevents access of the enzymes to susceptible sites of the cellulose molecule. It is postulated that the altered substrate consists of cellulose molecules of low degrees of polymerization, with some ring openings and altered chain ends. Weathered cellulose fails to adsorb cellulases. PMID- 16349884 TI - Pseudomonads and achromobacters in the spoilage of irradiated haddock of different preirradiation quality. AB - The effect of initial quality of fish on postirradiation (100 krad) changes in the bacterial flora of haddock fillets during aerobic storage at 3 C has been investigated, with emphasis on the Pseudomonas and Achromobacter groups. The quality was related to the length of time the eviscerated fish had been stored in ice prior to filleting. Increased numbers of organisms, in particular Pseudomonas putrefaciens, were found initially on fillets cut from older fish. Pseudomonads were reduced by 2 to 3 log orders by irradiation, and achromobacters and gram positive isolates predominated in the immediate postirradiation flora. Little difference could be detected in either types or relative proportions of organisms occurring during storage of unirradiated fish of different quality. Pseudomonads outgrew achromobacters and dominated the spoilage flora in all cases. After spoilage, however, the growth rate of pseudomonads declined markedly. In irradiated fish, achromobacters predominated throughout storage. In fish of better initial quality, bacterial numbers were 1 to 2 log orders higher at spoilage than in their unirradiated counterparts and in the poorer quality of irradiated samples. The increased number of organisms was accompanied by a radical change in the character of the predominant achromobacters. Pseudomonads were found to increase in numbers during storage of irradiated fish, in particular in poorer quality fish on which they were initially present in higher numbers. Detection of pseudomonads, even when present in high numbers, was found to be limited by the identification techniques normally used. PMID- 16349885 TI - Absence of 5-Hydroxy-4-Ketohexanoate and the alpha-Ketoglutarate Dehydrogenase Complex in Mutants of Saccharomyces oviformis Incapable of Growing on Ethanol. AB - The roles of the enzyme which forms 5-hydroxy-4-ketohexanoate (HKH) and of related enzymes in the metabolism of ethanol were studied in Saccharomyces oviformis WH92 and its mutants, which grew poorly or not at all on ethanol. The strains, which did not grow on ethanol, did not form HKH from alpha-ketoglutarate and acetaldehyde enzymatically and were also devoid of the alpha-ketoglutarate dehydrogenase complex. Acetaldehyde inhibited the activity of alpha-ketoglutarate dehydrogenase. These mutants did not grow on acetate since they had no acetyl-CoA synthetase activity. The relationship of the formation of HKH with the metabolism of ethanol is discussed. PMID- 16349887 TI - Isolation of fluorescent pseudomonads with a selective medium. AB - Incorporation of novobiocin, penicillin, and cycloheximide into a standard medium for fluorescence selects for fluorescent pathogenic and free-living pseudomonads. PMID- 16349888 TI - Microbial aromatization of androsta-1, 4, 7- triene-3, 17-Dione. AB - Several microorganisms transform androsta-1, 4, 7-triene-3, 17-dione into equilin and equilenin, presumably via a 19-hydroxylated intermediate. PMID- 16349886 TI - Enumeration of byssochlamys and other heat-resistant molds. AB - Methods for the detection of low numbers of heat-resistant molds on fruits were studied by using cultures of Byssochlamys and a number of unidentified mold isolates. Ascospore dormancy had a marked effect on viable recoveries, and the medium in which ascospores were heated influenced activation rates. Best results were obtained when fruit homogenates were heated for 60 min at 70 C in Concord grape juice, followed by culturing on acidified Potato Dextrose Agar. PMID- 16349889 TI - Improved tissue and cell homogenizer. AB - Modification of an existing tissue homogenizer is described, which converts to a milk-like consistency even difficult to macerate tissues such as cartilage, hair, and small bones. PMID- 16349892 TI - Rapid assay for bactericidal antibodies. AB - [This corrects the article on p. 279 in vol. 20.]. PMID- 16349893 TI - Stimulation of malo-lactic fermentation in eastern grape musts. AB - Induced malo-lactic fermentation was stimulated in Eastern grape musts by the addition of a new fermentation enhancer product. PMID- 16349890 TI - Whole microorganisms studied by pyrolysis-gas chromatography-mass spectrometry: significance for extraterrestrial life detection experiments. AB - Pyrolysis-gas chromatography-mass spectrometric studies of two microorganisms, Micrococcus luteus and Bacillus subtilis var. niger, indicate that the majority of thermal fragments originate from the principal classes of bio-organic matter found in living systems such as protein and carbohydrate. Furthermore, there is a close qualitative similarity between the type of pyrolysis products found in microorganisms and the pyrolysates of other biological materials. Conversely, there is very little correlation between microbial pyrolysates and comparable pyrolysis studies of meteoritic and fossil organic matter. These observations will aid in the interpretation of a soil organic analysis experiment to be performed on the surface of Mars in 1975. The science payload of this landed mission will include a combined pyrolysis-gas chromatography-mass spectrometry instrument as well as several "direct biology experiments" which are designed to search for extraterrestrial life. PMID- 16349894 TI - Susceptibility of Pseudomonas aeruginosa to Carbenicillin. AB - [This corrects the article on p. 630 in vol. 20.]. PMID- 16349895 TI - Rapid semiquantitative method for screening large numbers of virus samples by negative staining electron microscopy. AB - [This corrects the article on p. 261 in vol. 20.]. PMID- 16349897 TI - Preservation of Borrelia kansas and Plasmodium berghei. AB - [This corrects the article on p. 224 in vol. 20.]. PMID- 16349896 TI - Glycerol Ester Hydrolase Activity of Microbacterium thermosphactum. AB - Microbacterium thermosphactum possesses a significant glycerol ester hydrolase (lipase, EC 3.1.1.3) activity and a weak but definite carboxylic ester hydrolase (esterase, EC 3.1.1.1) activity. Harvested whole cell preparations contained 53 units of lipase activity with tripropionin as the substrate. This activity decreased with an increasing chain length of fatty acid in the triglyceride to 13 units with trilaurin as the substrate and no activity with tripalmitin. Maximum lipase activity was found at a temperature of 35 to 37 C and at a pH of 7.1 to 7.3. Lipase activity was associated with three different protein peaks when the protein of cell-free extract was fractionated by polyacrylamide gel electrophoresis. PMID- 16349898 TI - Virus in water: I. A preliminary study on a flow-through gauze sampler for recovering virus from waters. AB - [This corrects the article on p. 405 in vol. 21.]. PMID- 16349899 TI - Identification of members of the family enterobacteriaceae by the R-B system. AB - [This corrects the article on p. 880 in vol. 20.]. PMID- 16349900 TI - Some Factors Which Affect Amino Acid Uptake by Saccharomyces carlsbergensis. AB - When fully grown cells of Saccharomyces carlsbergensis were suspended in a solution of glucose and labeled amino acids, there was a lag phase before rapid uptake of certain amino acids. During this lag, significant amounts of sugar were utilized. The lag phase varied in length, depending upon the amino acid under study, but could be shortened by aeration of the cells and eliminated by their preincubation in glucose solution. Divalent metal ions, especially Ca added during the early stages of the lag phase, increased the length of the lag, an effect that could be reversed by washing with ethylenediaminetetraacetate, but amino acids which normally showed little or no lag before uptake were insensitive to Ca. The rate of uptake of amino acids or of sugar was essentially unaffected by Ca, whereas 2,4-dinitrophenol caused an overall decrease in the rate of uptake of all amino acids tested. The relevance of these observations to commercial brewing practice is shown. PMID- 16349901 TI - Detection of sulfa drugs and antibiotics in milk. AB - A disc assay method for testing sulfa drugs and antibiotics in milk was developed wherein Bacillus megaterium ATCC 9855 was used as the test organism and Mueller Hinton agar was used as the test substrate. Incubation was at 37 C for 4 to 5 hr. The test procedure is an improvement over the Bacillus subtilis-Antibiotic Medium No. 1 method, as described in Standard Methods for the Examination of Dairy Products, in that it is sensitive to eight sulfa drugs and to bacitracin without a significant change in sensitivity to eight other antibiotics commonly used for mastitis therapy. PMID- 16349902 TI - "Spore plate method" for transformation of steroids by fungal spores entrapped in silica gel g. AB - A new technique for investigating steroid biotransformations involving the use of glucose-treated Silica Gel G thin-layer chromatography plates spotted with fungal spores and steroid substrates is described. The conversion is followed by the detection and identification of steroid metabolites and is carried out on single plates by using the spores of different fungi. During the entire process, the spores remain on the original spots and microscopical examination revealed no germination. The method was successfully applied to as little as 30 mug of substrates, and a single plate could be used to detect the steroid metabolizing activity of spores of as many as 15 different cultures. PMID- 16349903 TI - Dry-heat resistance of bacterial spores recovered from mariner-Mars 1969 spacecraft. AB - The dry-heat resistances of 70 bacterial spore isolates recovered from Mariner Mars 1969 spacecraft were determined and expressed as D values (decimal reduction times). Fifty per cent of the spore isolates had D values of 60 min or less at 125 C. Of organisms with D values greater than 60 min, four were selected for a study of the effect of sporulation medium and suspension menstruum on dry-heat resistance. Both sporulation medium and suspension menstruum were found to affect significantly the dry-heat resistance of the bacterial spores tested. PMID- 16349904 TI - Relative frequency distribution of d(125 C) values for spore isolates from the mariner-Mars 1969 spacecraft. AB - Bacterial spore crops were prepared from 103 randomly selected aerobic mesophilic isolates collected during a spore assay of Mariner-Mars 1969 spacecraft conducted by the Jet Propulsion Laboratory. D(125 c) values, which were determined by the fractional-replicate-unit-negative-most-probable number assay method using a forced air oven, ranged from less than 5 min to a maximum of 58 min. Subsequent identification of the 103 isolates indicated that there was no relationship between species and dry-heat resistance. A theoretical dry-heat survival curve of the "population" was nonlinear. The slope of this curve was determined almost exclusively by the more resistant organisms, although they represented only a small portion of the "population." PMID- 16349905 TI - Growth of Aspergillus repens in Flue-Cured Tobacco. AB - In laboratory tests, flue-cured tobacco inoculated with Aspergillus repens was stored at 75, 80, 85, 87, and 95% relative humidity at 20 and 30 C. Samples were taken weekly for 4 weeks and evaluated for mold growth (colony count) and moisture content (MC). The weekly rate of fungus increase was slower at 20 C than at 30 C. Tobacco at 20 C with MC between 25 to 30% supported a slight to moderate increase in A. repens after 3 weeks of storage. However, tobacco at the same MC stored at 30 C was subject to rapid invasion by the fungus in as few as 1 to 2 weeks. Tobacco with MC above 30% stored at either 20 or 30 C became moldy in about 1 week. A mold index is proposed for evaluating populations of A. repens in tobacco. PMID- 16349906 TI - Microbiological spoilage of mayonnaise and salad dressings. AB - Saccharomyces bailii was isolated from two-thirds of the spoiled mayonnaise and salad dressing samples examined. Most of the rest were spoiled by Lactobacillus fructivorans. However, one sample contained large numbers of both S. bailii and L. plantarum. Two of the spoiled samples also contained small numbers of bacilli. Bacillus subtilis, B. pumilis, B. polymyxa, B. megaterium, and B. licheniformis were found in one sample and B. subtilis and B. pumilis in another. Small numbers of B. subtilis and B. licheniformis were also present in one unspoiled sample. Several media were evaluated for the isolation of L. fructivorans. S. bailii and L. fructivorans vigorously fermented glucose. The concentration of glucose in the spoiled samples ranged from 0 to 38.5 g/kg and from 1.3 to 17.8 g/kg for the unspoiled samples. PMID- 16349907 TI - Elutriation and Coulter Counts of Tetrahymena pyriformis Grown in Peanut and Cottonseed Meal Media. AB - Growth of Tetrahymena pyriformis W has been used to evaluate nutritional quality of peanut and cottonseed meals. An efficient elutriation method is described for separating cells of this organism from particulate matter left in the substrate (enriched with basal medium) after 4 days of incubation. After elutriation the cells can be counted with a Coulter counter by using calibration procedures which are presented. Elutriation and Coulter counting provide a rapid and efficient method of measuring the growth response of T. pyriformis W. Utility of the method is demonstrated by agreement between Coulter counts and visual counts of the cells and by demonstration of a linear response of cell numbers to substrate nitrogen. PMID- 16349908 TI - Microbial Flora of Pond-Reared Brown Shrimp (Penaeus aztecus). AB - Agar plate counts and microbial types are reported for brown shrimp reared in 2 acre natural marshland and in 0.5-acre artificial ponds during June to October 1970. Bacterial counts of pond-reared shrimp ranged from 5 x 10 to 5.5 x 10 per g. At final harvest in October, bacterial counts ranged from 2 x 10 to 5.5 x 10 per g. In marsh ponds, bacterial counts of shrimp and pond water were lowest in August when both water temperature and salinity were high. Coryneform bacteria and to a lesser extent Vibrio were the predominant isolates from fresh pond shrimp. Shrimp stored at 3 to 5 C for 7 days were acceptable as judged by appearance and odor. Between 7 and 14 days of refrigerated storage, bacterial counts increased sharply and about 50% of the samples became unacceptable. Refrigerated storage of pond shrimp caused increases in coryneform bacteria and micrococci and decreases in Vibrio, Flavobacterium, Moraxella, and Bacillus species. Pseudomonas species were not significant in fresh or stored pond shrimp. The microbial flora of pond water usually was dominated by coryneform bacteria, Flavobacterium, Moraxella, and Bacillus species. PMID- 16349909 TI - Adenosine Triphosphate Content of Selenastrum capricornutum. AB - The adenosine triphosphate content of Selenastrum capricornutum was maintained within 1.4 to 3.4 mug of adenosine triphosphate/mg (dry weight) of living biomass at different growth stages in media of different phosphorus concentrations. PMID- 16349911 TI - New Quantitative, Qualitative, and Confirmatory Media for Rapid Analysis of Food for Clostridium perfringens. AB - [This corrects the article on p. 501 in vol. 21.]. PMID- 16349910 TI - Chemically Defined Medium for Growth of Clostridium thermocellum, a Cellulolytic Thermophilic Anaerobe. AB - Nutritional requirements of Clostridium thermocellum were examined, and a defined medium was formulated which supported reproducible growth through 10 serial subcultures. PMID- 16349912 TI - Virus in water: I. A preliminary study on a flow-through gauze sampler for recovering virus from waters. AB - [This corrects the article on p. 405 in vol. 21.]. PMID- 16349913 TI - Role of carbon and nitrogen sources in bacterial growth and sporulation. AB - [This corrects the article on p. 131 in vol. 22.]. PMID- 16349914 TI - Production of Anticapsin by Streptomyces griseoplanus. AB - [This corrects the article on p. 1076 in vol. 21.][This corrects the article on p. 1077 in vol. 21.][This corrects the article on p. 1078 in vol. 21.][This corrects the article on p. 1079 in vol. 21.]. PMID- 16349915 TI - Survey of interferon production and sensitivity in human cell lines. AB - [This corrects the article on p. 102 in vol. 22.]. PMID- 16349916 TI - Fungal leaching of titanium from rock. AB - Penicillium simplicissimum solubilized up to 80% of the titanium in granitic rocks but less than 2% of the titanium in basaltic rocks. PMID- 16349917 TI - Decontamination of microbiological incubators. AB - A practical method of employing moist heat for the elimination of bacterial and fungal contaminants in microbiological incubators is described. PMID- 16349918 TI - Effect of flaming cotton-plugged tubes upon the contamination of media during culture transfers. AB - The effect of flaming the mouth of cotton-plugged broth tubes upon the contamination of media during culture transfers was studied. It was shown that flaming did not reduce contamination significantly either under usual laboratory conditions or when the laboratory atmosphere had been seeded with a test organism so as to achieve higher rates of colony-forming settling particles. PMID- 16349920 TI - Improved method for virus structural polypeptide analysis on dissociating acrylamide gel. AB - [This corrects the article on p. 760 in vol. 22.]. PMID- 16349919 TI - Microflora of soil as viewed by transmission electron microscopy. AB - Several procedures were evaluated for separating and concentrating indigenous microorganisms from soil without the occurrence of growth. Electron microscopy of nontangential, thin sections through these cells revealed that all of the cells examined were less than 0.9 mum in diameter, and up to 72% were "dwarf" cells less than 0.3 mum in diameter. Some were small enough that they should not be resolved with the light microscope. Approximately 27% had a fine structure bearing some resemblance to that of a bacterial cyst or microcyst, but this value may be low because cells having their outer layers partially stripped off were not included in the count. Approximately 25% showed a distinct periplasmic space, which often contained stainable material. Other fine structure features are presented together with frequencies of occurrence for the populations examined. PMID- 16349921 TI - Antimicrobial protection of moisturized deglet noor dates. AB - The growth of Saccharomyces rouxii and Saccharomyces mellis, which are two of the main spoilage organisms of dates, can be inhibited by various treatments. The most effective treatment found in this study that did not affect flavor consisted of a predip of the dates in 2% potassium sorbate solution followed by injection of methyl bromide into the sealed package. PMID- 16349922 TI - Ammonium, nitrate, and total nitrogen in the soil water of feedlot and field soil profiles. AB - A level feedlot, located in an area consisting of Wann silt loam changing with depth to sand, appears to contribute no more NO(3) nitrogen, NH(4) nitrogen, and total nitrogen to the shallow water table beneath it than an adjacent cropped field. Soil water samples collected at 46, 76, and 107 cm beneath the feedlot surface generally showed NO(3) nitrogen concentrations of less than 1 mug/ml. During the summer months, soil water NO(3) nitrogen increased at the 15-cm depth, indicating that nitrification took place at the feedlot surface. However, the low soil water NO(3) nitrogen values below 15 cm indicate that denitrification takes place beneath the surface. PMID- 16349923 TI - Purification of Pseudomonas aeruginosa Endotoxin by Membrane Partition Chromatography. AB - [This corrects the article on p. 211 in vol. 23.]. PMID- 16349925 TI - Use of aromatic compounds for growth and isolation of zoogloea. AB - [This corrects the article on p. 524 in vol. 23.]. PMID- 16349926 TI - Tape fermentation. AB - Microorganisms isolated from ragi, originally obtained from Indonesia, were selected for their ability to convert steamed glutinous rice into tape, an Indonesian fermented food. A mixture of Chlamydomucor oryzae and Endomycopsis fibuliger had good fermentation characteristics. Prepared starters, produced by growing pure cultures on rice and drying them, were as active as pure cultures grown for 4 days on Difco mycological agar slants at 30 C. PMID- 16349927 TI - Stomaching: a new concept in bacteriological sample preparation. AB - An entirely new mixing device, particularly suitable for preparing bacterial suspensions from foods, fabrics, swabs, and other fairly soft materials, has been developed. With this technique the sample and diluent are put into an inexpensive, sterile plastic bag which is vigorously pounded on its outer surfaces by paddles when placed inside the machine. The resulting compression and shearing forces effectively remove even deep-seated bacteria. After samples are taken for analysis the bag and its remaining contents are thrown away. Labor involved in cleaning and sterilizing reusable homogenizer cups or probes is eliminated, and the device is immediately ready for reuse. Running costs are thus drastically reduced, compared with conventional homogenizers. Additional advantages of this device, which is simple and inexpensive to manufacture, are low noise level, negligible temperature rise, and the small storage space required for bags. PMID- 16349928 TI - Apparatus for fixing, staining, and rinsing of tissue cultures for fluorescent antibody testing. AB - A staining tray and tray-housing container have been developed to facilitate fluorescent-antibody staining of tissue cultures on cover slips, which allows fixing, staining, and rinsing with a minimum of handling. Breakage and loss of cells were negligible. PMID- 16349929 TI - Demonstration of rhodanese activity in polyacrylamide gels. AB - Rhodanese activity from crude extracts of Thiobacillus sp. strain IV-85 was demonstrated in polyacrylamide gels after incubation in the reaction mixture by staining with dichloroindophenol in the presence of methylphenazonium methosulfate. The sensitivity of the staining system was found to be 8 x 10 moles of sulfite. PMID- 16349930 TI - Elemental analysis of vitamin-free casamino acids. AB - The elemental composition of three lots of vitamin-free Casamino Acids (Difco) was determined using X-ray fluorescence, atomic absorption, and colorimetric techniques. PMID- 16349931 TI - Microbial population of feedlot waste and associated sites. AB - A quantitative determination was made every 2 months for a year of the microflora of beef cattle waste and runoff at a medium-sized midwestern feedlot. Counts were obtained for selected groups of organisms in waste taken from paved areas of pens cleaned daily and, therefore, reflect the flora of raw waste. Overall, in terms of viable count per gram dry weight, the feedlot waste contained 10 total organisms, 10 anaerobes, 10 gram-negative bacteria, 10 coliforms, 10 sporeformers, and 10 yeasts, fungi, and streptomycetes. The specific numbers and pattern of these groups of organisms varied only slightly during the study in spite of a wide variation in weather. Data indicate that little microbial growth occurs in the waste as it exists in the feedlot. Runoff from the pens contained the same general population pattern but with greater variation attributable to volume of liquid. Comparable determinations of an associated field disposal area (before and after cropping), stockpiled waste, and elevated dirt areas in the pens indicate that fungi, and especially streptomycetes, are the aerobic organisms most associated with final stabilization of the waste. Yeasts, which are the dominant type of organism in the ensiled corn fed the cattle, do not occur in large numbers in the animal waste. Large ditches receiving runoff and subsurface water from the fields have a population similar to the runoff but with fewer coliforms. PMID- 16349932 TI - Enterobacteria in feedlot waste and runoff. AB - Samples of beef cattle feedlot waste (FLW), runoff from the pens, and water from a large drainage ditch at the feedlot were examined for Enterobacteriaceae. The drainage ditch receives the runoff but contains primarily subsurface drainage from fields on which FLW is spread for disposal. Planting and enrichment techniques with seven different media were used to isolate 553 cultures of enterobacteria. FLW contains about 50 million enterobacteria/g dry weight. More than 90% of these were Escherichia coli, none of which were enteropathogenic types as determined with multivalent sera. Citrobacter and Enterobacter cloacae were other organisms present in moderate numbers. Application of enrichment techniques broadened the spectrum of enterobacteria isolates to include the four Proteus spp., both Providencia spp., Klebsiella, Enterobacter aerogenes, Arizona, and a single isolate of Salmonella (serological group C(2)). Shigella was not isolated. The wide spectrum of enterobacteria in FLW may be a hazard if unsterilized waste is refed. Fewer enterobacteria occurred in the runoff and in the drainage ditch; the most numerous species in FLW also were most numerous at these sites. However, neither Salmonella nor Arizona was isolated from runoff or drainage-ditch waters. PMID- 16349933 TI - Hydrodynamic focusing and electronic cell-sizing techniques. AB - The technique of hydrodynamic focusing, used to improve the resolution of the Coulter counter for the sizing of bacteria, was examined. Latex particles of 0.26 mum to 6.7 mum volume were used to examine the characteristics of the system with and without hydrodynamic focusing. The system then was evaluated for sizing mixed bacterial populations as well as single populations. Possible applications are also discussed. PMID- 16349934 TI - Simple Method for Mass Production and Collection of Conidia from Hemispora stellata. AB - A simple method is described for the mass production and collection of conidia from Hemispora stellata. PMID- 16349935 TI - Improved cell dispersal method by low-temperature incubation of tissue in a trypsin-citrate solution. AB - Tissues soaked at low temperature in a trypsin-citrate solution, washed to remove enzymes, and disrupted by rapid mechanical vibrations gave yields of viable cells 1.6 to 5 times those from conventional procedures. PMID- 16349936 TI - Negligible evaporation retardation by oxyethylene docosanol under static conditions. AB - Oxyethylene docosanol (OED) retarded evaporation of agar media in petri plates when the plates were used in an air sampler; this confirmed the report of May (2). Flooding of plates with OED solutions or incorporation of OED into agar media, however, did not alter evaporation rates when the plates were stored in incubators or were refrigerated. PMID- 16349939 TI - Tape fermentation. AB - [This corrects the article on p. 976 in vol. 23.][This corrects the article on p. 977 in vol. 23.][This corrects the article on p. 978 in vol. 23.]. PMID- 16349937 TI - Novel system for improved control of filamentous microorganisms in continuous culture. AB - We describe a system devised to overcome the tendency of fungal mycelium grown in continuous cultures to adhere to the walls and hamper the outflow of the fermentation vessel, with resulting difficulties in attainment of true steadystate conditions. PMID- 16349940 TI - Death of Lactobacillus bulgaricus Resulting from Liquid Nitrogen Freezing. AB - Concentrated cell suspensions of Lactobacillus bulgaricus prepared from cells grown in semisynthetic media were frozen in liquid nitrogen. After storage for 24 hr, the cell suspensions were found to have decreased colony counts and acid producing capacity in milk. The amount of loss varied among the different strains tested. The addition of known cryoprotective agents to cell suspensions of the most labile strain before freezing provided little or no protection to the cells. However, storage stability of all strains investigated was improved by supplementing the growth medium with Tween 80 (polyoxyethylene sorbitan monooleate). The concentration of Tween 80 necessary for maximal storage stability varied among strains. PMID- 16349941 TI - 2,2-dibromo-3-nitrilopropionamide, a compound with slimicidal activity. AB - Laboratory and field tests demonstrated that 2,2-dibromo-3-nitrilopropionamide was an effective slimicide for use in papermaking systems and cooling towers. It was also effective as a bactericide for soluble oil emulsions. Acute toxicity tests showed that its hydrolysis at pH 9 and 23 C yielded products that were relatively nonhazardous to fathead minnows. PMID- 16349942 TI - Automated complement fixation with low reagent consumption. AB - [This corrects the article on p. 208 in vol. 24.]. PMID- 16349944 TI - Device for Eliminating Oil Condensation in the Air Flowmeter in CO(2) Incubator. PMID- 16349943 TI - Automatic surface inoculation of agar trays. AB - A machine is described which automatically inoculates a plastic tray containing agar media with a culture by use of either a conventional inoculating loop or a cotton swab. Isolated colonies were obtained with an inoculating loop when a heavy inoculum (10 cells/ml) was used or with a cotton swab when a light inoculum (ca. 10 cells/ml) was used. Trays containing combinations of differential or selective media were used to (i) separate mixtures of gram-positive and gram negative bacteria, (ii) facilitate isolation of organisms from clinical specimens, and (iii) compare colony growth characteristics of pure cultures. The design of the machine is simple, it is easy to use, and it relieves the operator from the manual task of streaking cultures. PMID- 16349945 TI - Reliable procedure for silica gel preparation. AB - A simple and reliable method is described for preparation of silica gel media for Nitrosomonas and Nitrobacter species. PMID- 16349946 TI - Activation of a Toxic Component of Clostridium botulinum Types C and D by Trypsin. AB - [This corrects the article on p. 108 in vol. 24.]. PMID- 16349947 TI - Polyelectrolyte flocculation as an aid to recovery of enteroviruses from oysters. AB - [This corrects the article on p. 540 in vol. 24.]. PMID- 16349948 TI - Potato aroma of lamb carcasses. AB - Rejection of lamb carcasses due to a strong potato odor was caused by Pseudomonas taetrolens. Although of rare occurrence, this defect can be of considerable commercial concern. PMID- 16349949 TI - Sauerkraut waste: a favorable medium for cultivating yeasts. AB - Sauerkraut waste was found to be more favorable for cultivating yeasts than malt extract broth and as good or better than peptone dextrose broth. PMID- 16349950 TI - Effect of Extracts of the Bark of Saccoglottis gabonensis on the Microflora of Palm Wine. AB - Direct addition of the bark of Saccoglottis gabonensis to fresh palm juice followed by microbial enumeration every 6 hr showed that the bark inhibited bacterial growth appreciably. The effect of various extracts of the bark on two bacterial species isolated from palm wine (Leuconostoc mesenteroides and Lactobacillus plantarum) was studied by using the paper disc method. It was observed that three of the five constituents of the bark showed inhibitory effect on the bacteria. PMID- 16349951 TI - Effect of low-roughage diets on the microflora and lipid metabolism in the rumen. AB - Changing the diet of five lactating cows and one nonlactating cow from high to low roughage induced milk fat depression in the lactating cows and altered the composition of the rumen microflora. While the numbers of lactic and propionic acid-producing bacteria increased, the numbers of Butyrivibrio spp. decreased. The numbers of lipolytic bacteria and the in vitro lipolytic activity of the rumen fluid were also decreased, as was the extent of hydrogenation of linoleic and linolenic acids combined in soybean oil incubated in vitro with rumen fluid. It is suggested that among the bacterial population in the rumen the vibrios, which were adversely affected by the low-roughage diets, may contribute significantly to both lipolysis and hydrogenation in the rumen. PMID- 16349952 TI - Agar medium for differential enumeration of lactic streptococci. AB - An agar medium containing arginine and calcium citrate as specific substrates, diffusible (K(2)HPO(4)) and undiffusible (CaCO(3)) buffer systems, and bromocresol purple as the pH indicator was developed to differentiate among lactic streptococci in pure and mixed cultures. Milk was added as the sole source of carbohydrate (lactose) and to provide growth-stimulating factors. Production of acid from lactose caused developing bacterial colonies to seem yellow. Subsequent arginine utilization by Streptococcus lactis and S. diacetilactis liberated ammonia, resulting in a localized pH shift back toward neutrality and a return of the original purple indicator hue. The effects of production of acid from lactose and ammonia were fixed around individual colonies by the buffering capacity of CaCO(3). After 36 hr at 32 C in a candle oats jar, colonies of S. cremoris were yellow, whereas colonies of S. lactis and S. diacetilactis were white. S. diacetilactis, on further incubation, utilized suspended calcium citrate, and, after 6 days, the citrate-degrading colonies exhibited clear zoning against a turbid background, making them easily distinguishable from the colonies of the other two species. The medium proved suitable for quantitative differential enumeration when compared with another widely used general agar medium for lactic streptococci. PMID- 16349953 TI - Associative growth studies in three-strain mixtures of lactic streptococci. AB - A recently developed differential agar medium was used to study associative growth patterns in 17 different heterologous, three-strain mixtures of Streptococcus lactis, S. cremoris, and S. diacetilactis grown in milk. Mixtures were made by combining equal volumes of 18-hr milk cultures of the three species. Relative populations of component species were followed through three successive transfers in milk after the initial mixed propagation. Direct evidence for strain dominance and compatibility was obtained. A procedure also was developed to estimate the extent of suppression of S. lactis and S. diacetilactis in a mixture containing a dominant S. cremoris strain. The technique described could be successfully applied in quality-control work in the dairy-starter manufacturing industry. PMID- 16349954 TI - An unclassified gram-negative rod isolated from the pharynx on thayer-martin medium (selective agar). AB - [This corrects the article on p. 772 in vol. 24.]. PMID- 16349955 TI - Casein as a necessary factor in the production of stimulatory material for associative growth of lactic streptococci. AB - Strains of Streptococcus cremoris KH and HC produced material that was stimulatory for S. cremoris R(6) in milk and in the dialyzable fraction of milk, but not in the dialysate fraction of milk, lactic acid whey, or lactose broth. The addition of casein to these latter media permitted the production of this stimulatory material to occur. Tryptone, peptone, and yeast extract could not be substituted for casein in producing the stimulatory material or in initiating associative growth in the lactic acid whey. The minimum concentration of casein required appeared to be from 2.0 to 2.5%. PMID- 16349956 TI - Production of yeast invertase from sauerkraut waste. AB - Sauerkraut waste was found to be a favorable medium for the production of invertase (beta-D-fructofuranoside fructohydrolase, EC 3.2.1.26) by Candida utilis. PMID- 16349957 TI - Tube-shaking attachment for rotary shakers. AB - An inexpensive tube-shaking device has been developed that attaches to rotary shakers without interfering with the flask holders already present. PMID- 16349958 TI - Heating device for funnels used in sterility testing. AB - A heating device is described for maintaining constant temperature in Pyrex funnels used during the performance of sterility tests. PMID- 16349960 TI - Introduction of Pseudomonas aeruginosa into a Hospital via Vegetables. AB - [This corrects the article on p. 567 in vol. 24.]. PMID- 16349959 TI - Effects of some disinfectants on african Swine Fever virus. AB - [This corrects the article on p. 115 in vol. 25.]. PMID- 16349961 TI - Method for Simultaneous Measurement of Radioactive and Inactive CO(2) Evolved from Soil Samples During Incubation with Labeled Substrates. AB - An apparatus is described which allows the simultaneous, continuous, and highly sensitive analysis of inactive and radioactive CO(2) evolved from C-supplemented soils or other materials. The apparatus consists of a control unit, a commercially available conductometric CO(2) analyzer, and fraction collector. A number of model experiments were conducted to demonstrate the potentials of the apparatus. These included analysis of the time course of priming action, when C glucose was added to soil, separation of CO(2) respiration peaks caused by simultaneous degradation of radioactive and inactive soil supplements, and study of the effects of a fungicide, Benomyl, on degradation of C-labeled glucose. In the last experiment, partial degradation of the fungicide could also be followed. PMID- 16349962 TI - Effects of natural microbial preparations on the electrokinetic potential of bacterial cells and clay minerals. AB - A complex mixture of fermentation residues and eutrophication products used commercially as a soil amendment and in various phases of sewage treatment was effective in reducing the electrophoretic mobility of clay minerals (kaolinite and montmorillonite) and cells of Agrobacterium radiobacter. The active fraction(s), which is active at very low concentrations, appears to be a stable (to heat, dialysis, concentration, and storage), net negatively charged polymer which may have several positively charged sites. The material does not significantly alter the viscosity or surface tension of aqueous systems and is probably a microbial metabolite(s). PMID- 16349963 TI - Determination of antibody to pneumoccal polysaccharides with chromic chloride treated human red blood cells and indirect hemagglutination. AB - [This corrects the article on p. 679 in vol. 24.]. PMID- 16349964 TI - Induction of Red Color Formation in Cabbage Juice by Lactobacillus brevis and Its Relationship to Pink Sauerkraut. AB - Membrane-filtered cabbage juice, when fermented by Lactobacillus brevis under conditions of controlled pH, frequently produced a water-soluble red pigment. The pigment, presumably responsible for imparting a highly objectionable discoloration to sauerkraut, was formed during the post logarithmic phase of growth. Color development is pH dependent (5.2 to 6.3) and can be suppressed by chemical reductants or anaerobic conditions of growth. The compound responsible for discoloration was purified and partially characterized. PMID- 16349965 TI - Effect of Frozen Storage Time on Heat Inactivation of Clostridium botulinum Type E Toxin. AB - [This corrects the article on p. 503 in vol. 25.]. PMID- 16349967 TI - Trends in wort carbohydrate utilization. AB - A gas chromatographic method suitable for any type of low-molecular-weight carbohydrate analysis has been utilized to determine the individual wort sugars in corn adjunct wort from a Western Canadian brewery. The fluctuations in each sugar during primary lager fermentation have been graphed. "End fermented" wort has been shown to contain some maltotriose, a small amount of maltose, and the nonfermentable carbohydrates, including maltotetraose, maltopentose, and dextrins. PMID- 16349968 TI - Novel Technique for Isolating Microstructures Present in Shake Cultures of the Fungus Ceratocystis ulmi. AB - Microstructures found in shake cultures of Ceratocystis ulmi, the fungus causing Dutch elm disease, have been isolated by a novel technique using the effect of bubbling gas through the culture filtrate. PMID- 16349969 TI - Culture of Ethiopian Strains of Borrelia recurrentis. AB - [This corrects the article on p. 935 in vol. 25.]. PMID- 16349970 TI - Modifications of the growth inhibition test and its application to human T mycoplasmas. AB - [This corrects the article on p. 528 in vol. 25.]. PMID- 16349971 TI - Isolation of Eikenella corrodens in a General Hospital. AB - [This corrects the article on p. 705 in vol. 25.]. PMID- 16349972 TI - Concentration of enteroviruses from large volumes of water. AB - An improved method for concentrating viruses from large volumes of clean waters is described. It was found that, by acidification, viruses in large volumes of water could be efficiently adsorbed to epoxy-fiber-glass and nitrocellulose filters in the absence of exogenously added salts. Based upon this finding, a modified version of our previously described virus concentration system was developed for virus monitoring of clean waters. In this procedure the water being tested is acidified by injection of N HCl prior to passage through a virus adsorber consisting of a fiber-glass cartridge depth filter and an epoxy-fiber glass membrane filter in series. The adsorbed viruses are then eluted with a 1 liter volume of pH 11.5 eluent and reconcentrated by adsorption to and elution from a small epoxy-fiber-glass filter series. With this method small quantities of poliovirus in 100-gallon (378.5-liter) volumes of tapwater were concentrated nearly 40,000-fold with an average virus recovery efficiency of 77%. PMID- 16349973 TI - Storage of brewing yeasts by liquid nitrogen refrigeration. AB - Many yeast strains are difficult to maintain in culture in a stable state, and long-term preservation by lyophilization, which has proved useful for other fungi, has given poor results with brewing yeasts. As an alternative to continuous subculture, which maximizes strain variability, various methods of cryogenic storage were investigated. Yeast strains were frozen with or without cryoprotectants (such as glycerol or inositol) and stored at -196 C. Recovery after warming was estimated from plate counts, and survivors were screened to detect changes in the frequency of morphological types, respiratory-deficient mutants, and glycerol-sensitive mutants. Strains varied in their sensitivity to freezing, and survival was modified by the growth medium, the freezing munstrua, and the freezing conditions. Suspension of cells in 10% (vol/vol) glycerol, cooled at 1 C/min, warmed rapidly and plated on malt-yeast extract-glucose peptone agar produced the highest percentage of viable colonies with a minimal change in metabolic characteristics. In two of the strains tested, no significant increase in mutation rate was detected under any of the treatments; the strains were maintained in a stable state and were metabolically comparable to unfrozen strains. In one strain of Saccharomyces uvarum after some freezing treatments, the percentage of respiratory-deficient mutants increased markedly, the fermentation rate declined, and a loss of flocculation occurred. The freezing parameters which increased the level of respiratory-deficient cells should be avoided in maintaining this strain. Maintenance of cultures of brewing yeasts by cryogenic storage has several advantages over other preservation techniques: the method is simple and reproducible, the cultures have remained stable over a 3 year test period, and the viability is high. PMID- 16349974 TI - Sporicidal properties of hydrogen peroxide against food spoilage organisms. AB - The sporicidal properties of hydrogen peroxide were evaluated at concentrations of 10 to 41% and at temperatures of 24 to 76 C. The organisms tested and their relative resistance at 24 C to 25.8% H(2)O(2) were: Bacillus subtilis SA 22 > B. subtilis var. globigii > B. coagulans > B. stearothermophilus > Clostridium sp. putrefactive anaerobe 3679 > S. aureus, with "D" values of 7.3, 2, 1.8, 1.5, 0.8., and 0.2 min, respectively. Heat shocking spores prior to hydrogen peroxide treatment decreased their resistance. Wet spores were more resistant than dry spores when good mixing was achieved during hydrogen peroxide treatment. Inactivation curves followed first-order kinetics except for a lag period where the inactivation rate was very slow. Increasing the H(2)O(2) concentration and the temperature reduced the lag period. PMID- 16349975 TI - Fungi that infect cottonseeds before harvest. AB - As a part of an investigation of aflatoxins and other mycotoxins in cottonseeds at harvest, samples of seeds collected from the 1971 crop at locations across the U.S. Cotton Belt were examined to determine the kinds of microorganisms causing internal or seed-coat infection in the field. Aspergillus flavus infection was absent from all seeds examined from most areas but was present in some samples from Arizona, California, and Texas. Fusarium spp., Alternaria sp., and A. niger caused internal infection at many locations; Colletotrichum gossypii and Rhizopus stolonifer were present in seeds from some areas but were generally much less common. Many of the infections with A. niger were in the seed coat. Bacterial infections were fairly frequent. In a series of commerical samples from Arizona. A. flavus infection was found in 61% of seeds, with fiber showing the bright, greenish-yellow (BGY) fluorescence that is diagnostic for A. flavus boll rot. Aflatoxin contamination was also concentration in the same seeds. The above findings agree with previous data showing that aflatoxin contamination of cottonseeds before harvest occurs rarely, if at all, in most parts of the U.S. Cotton Belt and that when such contamination does occur, it tends to be concentrated in seeds with the BGY fluorescence in their fiber and seed fuzz. PMID- 16349976 TI - Measurement of product temperature by use of an infrared thermometer. AB - An infrared sensor system to measure product surface temperature in industrial food processes is described. The method is suitable for use in either manufacturing plant or laboratory equipment. Good correlation can be expected between experimental and plant temperature readings. PMID- 16349977 TI - Effect of Nitrite and Nitrate on Toxin Production by Clostridium botulinum and on Nitrosamine Formation in Perishable Canned Comminuted Cured Meat. AB - [This corrects the article on p. 357 in vol. 25.]. PMID- 16349979 TI - Estimation of growth rate from the mitotic index. AB - [This corrects the article on p. 778 in vol. 25.]. PMID- 16349978 TI - Control of an estuarine microfouling sequence on optical surfaces using low intensity ultraviolet irradiation. AB - Ultraviolet light has been investigated as an active energy input for the control of slime film formation on optical surfaces submerged in San Francisco Bay for periods up to 6 weeks. Irradiation of quartz underwater windows was carried out from three positions: (i) exterior to the window, (ii) from directly behind the window, and (iii) from the edge of the window with the ultraviolet (UV) energy refracted through the front of the window. Internally administered irradiation reaching levels of 10 to 30 muW per cm measurable at the glass surface was effective in preventing bacterial slime film formation and settlement of metazoan larvae. When administered from the external position, over one order of magnitude more (500 to 600 muW/cm) UV energy was required to accomplish the same result. Irradiation from the edge position was most promising logistically and was effective in fouling control for 6 weeks. The results provide a preliminary quantitation of the energy requirement for control of the marine microfouling sequence which precedes development of macrofouling communities. PMID- 16349980 TI - Evaluation of a fluorescent antibody-enrichment serology combination procedure for the detection of salmonellae in condiments, food products, food by-products, and animal feeds. AB - [This corrects the article on p. 751 in vol. 25.]. PMID- 16349982 TI - Antibacterial effect of cysteine-nitrosothiol and possible precursors thereof. AB - [This corrects the article on p. 202 in vol. 27.]. PMID- 16349981 TI - Preparation and storage of high-titer lactic streptococcus bacteriophages. AB - Various techniques were employed for preparation of high-titer bacteriophage lysates of Streptococcus lactis, S. cremoris, and S. diacetilactis strains. Infection of a 4-h host culture in litmus milk at 30 C yielded the highest titers (2 x 10 to 4 x 10 plaque-forming units/ml) for most phages. Host infection in lactose-containing broth produced similar virus numbers only when 0.1 M tris(hydroxymethyl)aminomethane buffer stabilized the pH. The pH at the time of infection as well as the inoculum phage titer were critical in obtaining high titers. Optimum conditions for infection in broth were coupled with a polyethylene glycol concentration procedure to routinely produce milligram quantities of phage from 1 liter of lysate. Neutralization of whey lysates, as a means of storage, offered no survival advantage over unneutralized samples. Storage of phage lysates in a 15% glycerol whey solution at -22 C yielded a high rate of survival in most cases, even with repeated freezing and thawing, over a period of 24 months. PMID- 16349983 TI - Distribution of streptococcal groups in clinical specimens with evaluation of bacitracin screening. AB - [This corrects the article on p. 141 in vol. 27.]. PMID- 16349984 TI - Simple device for automatically transferring broth cultures. AB - A simple device is described for automatically transferring broth cultures from one tube to another by means of a tilting platform. PMID- 16349985 TI - Simple, inexpensive method for controlled freezing of cell cultures. AB - We have devised a system for controlled cooling of living cells which eliminates the need for complicated, expensive equipment but allows excellent recovery of viable cells after storage in liquid nitrogen. PMID- 16349987 TI - Effect of propylene oxide treatment on the microflora of pecans. AB - [This corrects the article on p. 768 in vol. 26.]. PMID- 16349986 TI - Use of plastic bags to maintain a humidified atmosphere for animal cell cultures. AB - A technique is described which allows incubation of petri dish tissue cultures in a dry incubator. PMID- 16349988 TI - Selective broth for isolation of group B streptococci. AB - [This corrects the article on p. 884 in vol. 26.]. PMID- 16349989 TI - Volatile Flavor Compounds Produced by Molds of Aspergillus, Penicillium, and Fungi imperfecti. AB - Strains of molds Aspergillus niger, A. ochraceus, A. oryzae, A. parasiticus, Penicillium chrysogenum, P. citrinum, P. funiculosum, P. raistrickii, P. viridicatum, Alternaria, Cephalosporium, and Fusarium sp. were grown on sterile coarse wheat meal at 26 to 28 C for 120 h. The volatiles from mature cultures were distilled at low temperature under reduced pressure. The distillates from traps -40 and -78 C were extracted with methylene chloride and subsequently concentrated. All the concentrates thus obtained were analyzed by gas-liquid chromatography, mass spectrometry, chemical reactions of functional groups, and olfactory evaluation. Six components detected in the culture distillates were identified positively: 3-methylbutanol, 3-octanone, 3-octanol, 1-octen-3-ol, 1 octanol, and 2-octen-1-ol. They represented 67 to 97% of all the volatiles occurring in the concentrated distillate. The following 14 components were identified tentatively: octane, isobutyl alcohol, butyl alcohol, butyl acetate, amyl acetate, octyl acetate, pyridine, hexanol, nonanone, dimethylpyrazine, tetramethylpyrazine, benzaldehyde, propylbenzene, and phenethyl alcohol. Among the volatiles produced by molds, 1-octen-3-ol yielding a characteristic fungal odor was found predominant. PMID- 16349990 TI - Microbial petroleum degradation: use of mixed hydrocarbon substrates. AB - Methods of examining hydrocarbons to estimate the microbial degradation of petroleum are compared. Gas-liquid chromatography with a mixed hydrocarbon substrate has been shown to be useful in evaluating microbial potential for degradation of a number of hydrocarbons. PMID- 16349991 TI - Cellulase Production by Trichoderma viride on Feedlot Waste. AB - Feedlot waste contains essentially all the necessary nutrients for batch fermentation with the fungus Trichoderma viride. The organism utilizes two-thirds of the carbohydrate in feedlot waste while elaborating cellulase in quantities comparable to commercial preparations. Essentially odor-free, the fermented waste contains all of the original nitrogen but has 24% less organic matter. PMID- 16349992 TI - Transformation of 1-Menthene by a Cladosporium: Accumulation of beta-Isopropyl Glutaric Acid in the Growth Medium. AB - An organism isolated from soil samples collected near a terpene plant grew in the presence of 1-menthene, with no other major source of carbon and energy. The organism was tentatively identified as a member of the genus Cladosporium. When this organism was grown in the presence of 1-menthene, with no other major source of carbon, substantial quantities of beta-isopropyl glutaric acid accumulated in the fermentation medium. beta-Isopropyl glutaric acid is probably an end product of the degradation of 1-menthene by the organism. PMID- 16349993 TI - Physical state of phenanthrene for utilization by bacteria. AB - Experiments were done to determine if bacteria able to utilize phenanthrene as a sole source of carbon obtain the hydrocarbon directly from solid phenanthrene particles suspended in the medium or from the phenanthrene dissolved in the medium. Growth experiments were done in gas-tight fermentors completely filled with air-saturated, mineral salts solution with phenanthrene as the substrate. Generation times were determined by rate of oxygen consumption. Generation times were independent of the amount of solid phenanthrene present, and these generation times were the same as the generation time on medium containing only dissolved phenanthrene. It was concluded that bacteria utilize phenanthrene in the dissolved state. PMID- 16349994 TI - Persistence of bacteriophages in experimentally infected cell cultures. AB - A bacteriophage, isolated from a live measles vaccine and designated varphiV 1(L), was intentionally introduced into cell culture systems. The bacterial virus persisted in the cell cultures after daily medium changes and trypsinization, and in cell lysates prepared by freezing and thawing. The results suggest that phages contaminating cell cultures may be carried over into the final product during the manufacture of viral vaccines. PMID- 16349995 TI - Identification of yeasts from the suwannee river Florida estuary. AB - The yeast flora of the Suwannee River estuary in Florida has been studied. The predominant genera were Candida and Rhodotorula; however, the yeast most frequently isolated was Cryptococcus laurentii. Nine ascosporogenous species were isolated, with Hansenula saturnus predominating. The salinity range of the sediments was 0.4 to 20.6%; in the estuary water, 0.07 to 0.25%; and in the open Gulf of Mexico, 18 to 20%. PMID- 16349996 TI - Thermophilic mixed culture of bacteria utilizing methanol for growth. AB - A thermophilic mixed population of bacteria, capable of utilizing methanol as its sole carbon-energy source at temperatures up to 65 C, was selected by enrichment and studied. A maximal cellular yield of 0.42 g per g of methanol was observed at 50 to 56 C. The maximal specific growth rate of the mixed population in continuous culture at 56 C was greater than 0.32 per h. The amino acid profile of the mixed culture indicated that a high quality protein was produced and the protein content was 71%. The properties of this culture and its ability to grow at elevated temperatures are discussed in terms of single-cell protein production and the treatment of industrial waste. PMID- 16349997 TI - Rumen bacterial interrelationships with plant tissue during degradation revealed by transmission electron microscopy. AB - The mode of rumen bacterial degradation of cell walls in coastal bermudagrass [Cynodon dactylon (L) Pers.] differed with the plant tissue type. Bacteria degraded thin, primary cell walls of mesophyll and phloem apparently by extracellular enzymes and without prior attachment; thick-walled bundle sheath and epidermal cells apparently were degraded after bacterial attachment, in some types by an extracellular substance, to the plant cell walls. Rumen bacteria split the nondegraded cuticle from the epidermis by preferentially attacking the cell just underneath the cuticle. The propensity for bacterial attachment to lignified cells of the vascular tissue was low, and bacterial degradation of these cells did not occur after 72 h of incubation. PMID- 16349998 TI - Evaluation of porcelain cup soil water samplers for bacteriological sampling. AB - The validity of obtaining soil water for fecal coliform analyses by porcelain cup soil water samplers was examined. Numbers from samples of manure slurry drawn through porcelain cups were reduced 100- to 10,000,000-fold compared to numbers obtained from the external manure slurry, and 65% of the cups yielded coliform free samples. Fecal coliforms adsorbed to cups apparently were released, thus influencing the counts of subsequent samples. Fecal coliforms persisted in soil water samplers buried in soil and thus could significantly influence the coliform counts of water samples obtained a month later. These studies indicate that porcelain cup soil water samplers do not yield valid water samples for fecal coliform analyses. PMID- 16349999 TI - Survival of Saccharomyces cerevisiae and Sarcina lutea at Refrigerator Temperatures. AB - Assay organisms Saccharomyces cerevisiae and Sarcina lutea, when suspended in a weak buffer and stored at refrigerator temperatures, are stable for 1 year or more. PMID- 16350000 TI - Simple, inexpensive method for controlled freezing of cell cultures. AB - [This corrects the article on p. 616 in vol. 27.]. PMID- 16350001 TI - Radioimmunoassay on polycarbonate membranes: a sensitive and simplified method for the detection and quantitation of antibody. AB - [This corrects the article on p. 475 in vol. 27.]. PMID- 16350002 TI - Bacteremia after tooth extractions studied with the aid of prereduced anaerobically sterilized culture media. AB - [This corrects the article on p. 931 in vol. 27.]. PMID- 16350003 TI - Rice-Grown Rhizopus oligosporus Inoculum for Tempeh Fermentation. AB - A method of growing Rhizopus oligosporus on cooked rice as the inoculum for the fermentation of soybeans into tempeh was described and evaluated. Isolated R. oligosporus spores on glass beads survived best at low temperature and intermediate humidity. The activity of the rice-grown inoculum to ferment soybeans into tempeh did not decrease appreciably when stored desiccated for one year at 4 C or room temperature. Bacterial contaminants as high as 10 counts per g of cooked soybeans did not seem to affect the fermentation. PMID- 16350004 TI - Monitoring the atmosphere in an anaerobic chamber. AB - The Couloximeter, a fuel cell designed to measure trace amounts of oxygen, was used to monitor the atmosphere in an anaerobic chamber. The device, easy to operate and to maintain, allowed both major and minor fluctuations in oxygen concentration to be measured. Using a hose attached to the outlet within the box, defective (ruptured) gloves were consistently distinguishable from intact gloves. PMID- 16350005 TI - Microbial response to drought in a Texas highplains shortgrass prairie. AB - The population of the microbial flora of a mixed blue gramma grass (Bouteloua gracilis H. B. K.) and prickly pear (Opuntia polyacantha Haw.) prairie near Amarillo, Texas, was studied during 1971 after a severe drought. Bacteria, fungi, and algae were estimated by plate count and terminal dilution procedures. Rates of grass and paper decomposition were determined. The microbial flora of soil associated with bovine-grazed grass did not differ significantly from the flora associated with ungrazed grass, either qualitatively or quantitatively. During drought, a greater number of fungi were found in soil associated with prickly pear than in that associated with blue gramma grass. The microbial biomass decreased one full log between the surface and a depth of 50 cm, and the percentage of anaerobes increased with depth. The maximum numbers of fungi and algae detected were 8 x 10 and 6 x 10/g respectively. A linear relationship existed between the microbial biomass and soil moisture. The maximum number of aerobic, heterotrophic bacteria detected was 1.5 x 10 viable cells per g of soil. PMID- 16350006 TI - Recovery of Lactobacillus bulgaricus and Streptococcus thermophilus on Nine Commonly Used Agar Media. AB - Of the nine media tested, Eugon, Elliker's lactic agar, pH 6.8, and modified tryptic soy broth agars showed superior recovery of Lactobacillus bulgaricus and Streptococcus thermophilus strains. PMID- 16350007 TI - Arthrobacter globiformis and Its Bacteriophage in Soil. AB - Bacteriophages in soil for Arthrobacter globiformis were rarely detected unless the soil was nutritionally amended and incubated. In amended soil, phage were continuously produced for at least 48 h, and this did not require the addition of host cells. Rod and spheroid stage host cells added to the amended soil encountered indigenous bacteriophage, but added phage did not encounter sensitive indigenous host cells for some time, if at all. The indigenous phage in nonincubated soil seemed to be present in a masked state which was not merely a loose physical adsorption to soil materials but required growth conditions other than lysogeny for them to increase their titers. The possibility is discussed that the indigenous host cells in nonamended soil are present in a nonsensitive spheroid state, with the cells becoming sensitive to the phage in a rate-limiting fashion as nonsynchronous outgrowth occurs for a portion of the spheroid cells. PMID- 16350008 TI - Microflora of four fermented fish sauces. AB - Thirty-nine microorganisms representing 11 species of bacteria, one yeast, and three filamentous fungi were isolated and identified from four fermented fish sauces: nampla, patis, koami, and ounago. PMID- 16350009 TI - Studies on batch production of bacterial concentrates from mixed species lactic starters. AB - Optimum growth conditions for mixed species starter FDs 0172 at constant pH in skim milk, whey, and tryptone medium were investigated. Growth rate and maximum population were optimal at 30 C. pH values between 5.5 and 7.0 did not influence the growth rate and maximum population obtainable. Lactic acid-producing activity declined rapidly after reaching the end of the exponential growth phase. The bacterial balance was found to be influenced by the growth parameters: media, pH, temperature, and neutralizer. Skim milk or whey medium at 25 C, pH 6.5, and neutralized with 20% (vol/vol) NH(4)OH kept the bacterial balance almost constant throughout the cultivation. Grown in tryptone medium at constant pH, the changes in bacterial balance and other metabolic activities were striking compared to the other two media tested. The effect of lactate as an inhibitor was found to be complex, changing with the growth conditions. Concentrates made from mixed species starters FDs 0172, FD 0570, CH 0170, CHs 0170, and T 27 were comparable to controls when cultivated at the optimum conditions found and thereafter centrifuged. Aroma production, proteolytic activity, and CO(2) production did not change significantly compared to controls when cultivated at optimum conditions in skim milk or whey medium. PMID- 16350010 TI - Ecology of soil arthrobacters in clarion-webster toposequences of iowa. AB - Toposequence variations in soil properties were characterized and related to variations in populations of total isolatable bacteria and arthrobacters. Increases in soil NO(3)-N, available phosphorous, NO(3)-N-producing power, Arthrobacter counts, and the percentage of the total counts represented by arthrobacters were correlated with decreases in soil acidity. The total bacterial counts were not correlated with soil acidity but were associated with percentage of soil organic matter and percentage of clay. The percentage of the total counts represented by arthrobacters was lowest at the summit position and increased downslope to the highest value in the toeslope position. Factor analysis of the data revealed that 67 to 81% of the total variance exhibited by all variables per site-sampling period could be accounted for by soil acidity, soil structure, soil fertility, soil moisture, and bacterial factors. A selective medium was developed for soil arthrobacters and tested on a wide variety of central Iowa soils to determine its potential as a medium for enumeration as well as isolation. The medium developed in this study was found to be superior to the other available direct-isolation media for soil arthrobacters. PMID- 16350012 TI - Effects of seawater cations and temperature on manganese dioxide-reductase activity in a marine bacillus. AB - [This corrects the article on p. 785 in vol. 28.]. PMID- 16350011 TI - Longevity of dehydrated violet red bile agar. AB - The length of storage in sealed containers at room temperature did not affect the plating efficiency of violet red bile agar. PMID- 16350013 TI - Studies on batch production of bacterial concentrations from mixed species lactic starters. AB - [This corrects the article on p. 133 in vol. 29.]. PMID- 16350014 TI - Detection and Growth of Enteropathogenic Escherichia coli in Soft Ripened Cheese. AB - [This corrects the article on p. 179 in vol. 29.]. PMID- 16350016 TI - Effect of environmental parameters on bacterial degradation of bunker C oil, crude oils, and hydrocarbons. AB - [This corrects the article on p. 915 in vol. 28.]. PMID- 16350015 TI - Growth of a Mixed Species Lactic Starter in a Continuous "pH-Stat" Fermentor. AB - Optimum growth conditions for mixed species starter FDs 0172 in skim milk, whey medium, and tryptone medium at 25 C in a "pH-stat" continuous fermentor were investigated. Specific growth rate and productivity were found to be influenced both by the medium and the pH value. Highest productivity was found in skim milk medium between pH 5.5 and 5.9. Lactic acid-producing activity of harvested cells decreased by continuous cultivation at pH values lower than 5.9 to 6.1, especially in tryptone and whey medium. Bacterial balance, CO(2) production, and aroma formation were comparable to the control in skim milk and whey medium at any pH, but differed significantly for cells grown in tryptone medium. PMID- 16350017 TI - Rumen bacterial degradation of forage cell walls investigated by electron microscopy. AB - The association of rumen bacteria with specific leaf tissues of the forage grass Kentucky-31 tall fescue (Festuca arundinacea Schreb.) during in vitro degradation was investigated by transmission and scanning electron microscopy. Examination of degraded leaf cross-sections revealed differential rates of tissue degradation in that the cell walls of the mesophyll and pholem were degraded prior to those of the outer bundle sheath and epidermis. Rumen bacteria appeared to degrade the mesophyll, in some cases, and phloem without prior attachment to the plant cell walls. The degradation of bundle sheath and epidermal cell walls appeared to be preceded by attachment of bacteria to the plant cell wall. Ultrastructural features apparently involved in the adhesion of large cocci to plant cells were observed by transmission and scanning electron microscopy. The physical association between plant and rumen bacterial cells during degradation apparently varies with tissue types. Bacterial attachment, by extracellular features in some microorganisms, is required prior to degradation of the more resistant tissues. PMID- 16350019 TI - Laboratory method for fermentation of meat and poultry sausages in fibrous casings. AB - The construction and operation of a relatively inexpensive cabinet for sausage fermentation studies is described. Temperature can be controlled to +/-1 C with a relative humidity of approximately 95%. PMID- 16350018 TI - Improved medium for lactic streptococci and their bacteriophages. AB - Incorporation of 1.9% beta-disodium glycerophosphate (GP) into a complex medium resulted in improved growth by lactic streptococci at 30 C. The medium, called M17, contained: Phytone peptone, 5.0 g; polypeptone, 5.0 g; yeast extract, 2.5 g; beef extract, 5.0 g; lactose, 5.0 g; ascorbic acid, 0.5 g; GP, 19.0 g; 1.0 M MgSO(4).7H(2)O, 1.0 ml; and glass-distilled water, 1,000 ml. Based on absorbance readings and total counts, all strains of Streptococcus cremoris, S. diacetilactis, and S. lactis grew better in M17 medium than in a similar medium lacking GP or in lactic broth. Enhanced growth was probably due to the increased buffering capacity of the medium, since pH values below 5.70 were not reached after 24 h of growth at 30 C by S. lactis or S. cremoris strains. The medium also proved useful for isolation of bacterial mutants lacking the ability to ferment lactose; such mutants formed minute colonies on M17 agar plates, whereas wild type cells formed colonies 3 to 4 mm in diameter. Incorporation of sterile GP into skim milk at 1.9% final concentration resulted in enhanced acid-producing activity by lactic streptococci when cells were inoculated from GP milk into skim milk not containing GP. M17 medium also proved superior to other media in demonstrating and distinguishing between lactic streptococcal bacteriophages. Plaques larger than 6 mm in diameter developed with some phage-host combinations, and turbid plaques, indicative of lysogeny, were also easily demonstrated for some systems. PMID- 16350020 TI - Violet red bile 2 agar for stressed coliforms. AB - [This corrects the article on p. 537 in vol. 29.]. PMID- 16350021 TI - Machine for printing hydrophobic grids on membrane filters. AB - An apparatus for printing wax grids on membrane filters is described. The machine produced more and better hydrophobic grid-membrane filters much more rapidly than by hand. PMID- 16350022 TI - Changes in microbial population during fermentation of feedlot waste with corn. AB - A new process for recycling feedlot waste involves the fermentation of liquid from this waste combined with corn. Changes in the flora of this silage-like fermentation were followed. The fermentation was dominated by lactobacilli and yeasts, which initially constitute 1% or less of the natural flora. The species of yeasts and lactics involved were characterized. The fermentation has two phases. A single heterolactic species multiplied rapidly for the first 24 h until it represented 95% of the lactobacilli and more than 90% of the total microflora. It displaced the betabacterium predominant among lactics of the original waste; the acid produced killed coliforms and other organisms in feedlot waste; and the acetic acid produced probably caused the death of the dominant native yeast Trichosporon cutaneum (de Beurm., Gougerot et Vaucher) Ota. The peak lactobacillus count remained constant (about 2 x 10 organisms/g [wet weight]) throughout the rest of the fermentation. Homolactics dominated the later phase and yeasts increased to 9.5 x 10 organisms/g (wet weight). At 6 days, a stable mixture of three lactobacilli was present, one streptobacterium, one thermobacterium, and one betabacterium. Similarly, yeasts stabilized as a mixture of two Candida sp. and one Pichia sp. The dominant species of lactics were characterized. Information on the sequence of microorganisms provides a basis for enhanced protein synthesis in the fermentation. PMID- 16350023 TI - Effects of Bicarbonate on Growth of Neisseria gonorrhoeae: Replacement of Gaseous CO(2) Atmosphere. AB - [This corrects the article on p. 469 in vol. 29.]. PMID- 16350024 TI - Isolation and Some Properties of Glucoamylase from Cephalosporium charticola Lindau. AB - High glucoamylase (alpha-D-/1 --> 4/glucan glucohydrolase, EC 3.2.1.3.) activity was obtained in the cell-free culture fluid of Cephalosporium charticola. Glucoamylase seems to be the only amylolytic enzyme produced by C. charticola. The enzyme, purified on diethylaminoethyl-cellulose, was homogeneous by disc gel electrophoresis. The optimum pH on starch was 5.4, and optimum temperature was 60 C. Starch was degraded more rapidly than several other substrates; maltose was hydrolyzed about one-fifth as rapidly as starch. The molecular weight was 69,000, as determined by Sephadex G-100 filtration. The enzyme is a glycoprotein and contains about 6.6% sugars (mannose and glucosamine). PMID- 16350025 TI - Neutral Lipids and Phospholipids of Free-Living and Bacteroid Forms of Two Strains of Rhizobium Infective on Lotus pedunculatus. AB - The neutral lipids and phospholipids of two strains of rhizobia in their free living state and in symbiosis with a host plant are described. The principal lipid classes found were the polymer poly-beta-hydroxybutyrate, phospholipids, free fatty acids, glycerides, methyl esters, aliphatic alcohols, and hydrocarbons. The lipids include unusual unsaturated methyl-branched and saturated methoxy-branched fatty acids. Most components were found to be common to both forms of both strains, although the proportions varied. A number of strain differences could be discerned. PMID- 16350026 TI - Studies on thermophilic cellulolytic fungi. AB - Three thermophilic cellulolytic fungi, Chaetomium thermophile var. coprophile, Sporotrichum thermophile, and Thermoascus aurantiacus were studied to determine the conditions for a high rate of cellulose degradation. The range of temperature over which good growth occurred was determined first in a temperature gradient incubator; the optimum temperature was then established in shake flask cultures. T. aurantiacus had the highest optimum growth temperature range (46 to 51 C), whereas S. thermophile had the broadest range over which good growth occurred (36 to 43 C). Optimum temperatures for the three organisms, T. aurantiacus, S. Thermophile, and C. thermophile were 48, 40, and 40 C, respectively. It was found that the addition of an organic carbon and nitrogen source to a cellulose mineral solution medium markedly increased the rate of cellulose degradation. The surfactant, Tween 80, which has been reported to be of value in the production and recovery of the enzyme, cellulase, was shown to be detrimental to the degradation of cellulose in culture. In the medium used, S. thermophile gave the highest rate of substrate utilization; 56% of the cellulose was hydrolyzed in 72 h. The average degree of polymerization of cellulose decreased from 745 to 575. PMID- 16350027 TI - Influence of substrate wettability on the attachment of marine bacteria to various surfaces. AB - The effect of the initial substrate surface condition, as indicated by the critical surface tension for wetting, on the rate of attachment of marine bacteria to a variety of solid surfaces has been measured. The techniques used to determine the number of bacteria attached per unit surface area were a lipopolysaccharide test utilizing Limulus lysate and direct examination of the surface by scanning electron microscopy. The results obtained by the two techniques are compared and their significance to the control of microbiological slime film formation (microfouling) is discussed. PMID- 16350028 TI - Microbial degradation of ethylenediaminetetraacetate in soils and sediments. AB - [C]ethylenediaminetetraacetate (EDTA) was shown to be slowly biodegraded to CO(2) in soils and sediments under aerobic conditions and by microorganisms in mixed liquid culture. EDTA chelates of Cu, Cd, Zn, Mn, Ca, and Fe added to soil were equally degraded, while Ni-EDTA was degraded more slowly. PMID- 16350029 TI - Automatic pH Control and Soluble and Insoluble Substrate Input for Continuous Culture of Rumen Microorganisms. AB - An artifical rumen continuous culture with pH control, automated input of water soluble and water-insoluble substrates, controlled mixing of contents, and a collection system for gas is described. PMID- 16350030 TI - Effects of a commercial malathion preparation on selected soil bacteria. AB - The aromatic petroleum distillates present in a commercial preparation of malathion insecticide had a greater effect on viability of pure cultures of soil bacteria than did the malathion, whereas neither component had an effect on the population of the natural soil community. PMID- 16350032 TI - Improved medium for lactic streptococci and their bacteriophages. AB - [This corrects the article on p. 807 in vol. 29.]. PMID- 16350031 TI - Persistence and biodegradation of spilled residual fuel oil on an estuarine beach. AB - [This corrects the article on p. 646 in vol. 29.]. PMID- 16350033 TI - Cellulase production by a thermophilic clostridium species. AB - Strain M7, a thermophilic, anaerobic, terminally sporing bacterium (0.6 by 4.0 mum) was isolated from manure. It degraded filter paper in 1 to 2 days at 60 C in a minimal cellulose medium but was stimulated by yeast extract. It fermented a wide variety of sugars but produced cellulase only in cellulose or carboxymethyl cellulose media. Cellulase synthesis not only was probably repressed by 0.4% glucose and 0.3% cellobiose, but also cellulase activity appeared to be inhibited by these sugars at these concentrations. Both C(1) cellulase (degrades native cellulose) and C(x) cellulase (beta-1,4-glucanase) activities in strain M7 cultures were assayed by measuring the liberation of reducing sugars with dinitrosalicylic acid. Both activities had optima at pH 6.5 and 67 C. One milliliter of a 48-h culture of strain M7 hydrolyzed 0.044-meq of glucose per min from cotton fibers. The cellulase(s) from strain M7 was extracellular, produced during exponential growth, but was not free in the growth medium until approximately 30% of the cellulose was hydrolyzed. Glucose and cellobiose were the major soluble products liberated from cellulose by the cellulase. ZnCl(2) precipitation appeared initially to be a good method for the concentration of cellulase activity, but subsequent purification was not successful. Isoelectric focusing indicated the presence of four C(x) cellulases (pI 4.5, 6.3, 6.8, and 8.7). The rapid production and high activity of cellulases from this organism strongly support the basic premise that increased hydrolysis of native cellulose is possible at elevated temperature. PMID- 16350034 TI - Characteristics of heavily mucoid bacterial isolated from fish pen slime. AB - Weakly gram-positive pleomorphic rods isolated from the slime on the storage pen surfaces of fishing trawlers were found to produce extensive capsular material and intensely mucoid colonies. The isolates studied produced highly pleomorphic club-shaped cells indicative of coryneforms. Their DNA base composition ranged from 64.2 to 68.2 mol% guanine plus cytosine on the basis of melting point determinations. PMID- 16350035 TI - Inhibition of microbial growth by Fatty amine catalysts from polyurethane foam test tube plugs. AB - [This corrects the article on p. 615 in vol. 29.]. PMID- 16350037 TI - Effect of incubation conditions at 55 C on moisture loss from agar plates. AB - The percentage of moisture loss was least from plates incubated in plastic bags, sealed jars, or in a humid chamber. There was a stacking effect noted in plates incubated in plastic bags and sealed jars. PMID- 16350036 TI - Factors affecting the contribution by epiphytic algae to the primary productivity of an oligotrophic freshwater lake. AB - A diatom-dominated population of epiphytic algae was studied in an oligotrophic lake to determine the factors which limit epiphyte growth and to measure their contribution to primary productivity. Algae were collected from plants growing at four sites in Lake George, N.Y., during the spring, summer, and fall of 1974. Samples were taken from 3 m, corresponding to the depth at which macrophytes were most productive. Algae exhibited an optimum temperature for HCO(3) uptake at 30 C, although the summer littoral lake temperature ranged from 18 to 25 C. Light saturation occurred at an intensity of 8,608 lux, approximating the environmental intensity at the depth from which algae were taken. Epiphytes exhibited their maximum photosynthetic capacity of 0.6 mg of carbon fixed/m of macrophyte surface area per h in the early afternoon in mid-August. They assimilated approximately 5% as much inorganic carbon as the macrophytes from which they were taken. Epiphyte population densities followed the seasonal growth patterns of the macrophytes, with maximal leaf colonization remaining essentially constant relative to the leaf position on the plant. There was little change in density between sampling sites at any given time. Productivities of epiphytes from bottom leaves were 10-fold greater than those of epiphytes from top leaves. Addition of PO(4), NO(3), NH(3), Si, and SO(4) had no stimulatory effect on photosynthesis. Addition of HCO(3) stimulated photosynthesis greater than 30%, suggesting that carbon may be a limiting nutrient for epiphytic algae in Lake George. PMID- 16350038 TI - Purification and Properties of Intracellular Proteinase from Streptococcus cremoris. AB - Proteolytic activity in the extract from the cells of Streptococcus cremoris increased in the presence of casein, lactose, glucose, and CaCl(2) in the media but was negligibly detectable in the extract of the cells harvested from the culture containing succinate or citrate. The intracellular proteinase from S. cremoris harvested from tomato medium was purified 150-fold in this experiment. The enzyme had a molecular weight of 140,000, optimum pH at 6.5 to 7.0, and maximum activity at 30 C. The proteinase was activated by Ca and inhibited by Zn, Cu, Hg, Fe, ethylenediaminetetraacetate, and sodium lauryl sulfate. The K(m) value of the enzyme towards each casein fraction was almost the same, and the V(max) of the enzyme towards alpha(s)-casein was smaller than those towards the other casein fractions. PMID- 16350039 TI - Dual channel temperature recorder. AB - A dual channel temperature recorder is described which can measure temperatures linearly over a range of 0 to 80 C. The sensitivity can be adjusted so that any temperature span, as small as 3 degrees , will cover recorder full scale, and temperatures can be read to 0.1 degrees . The recorder provides two independent, permanent records of temperature variations which can be related directly to the time of day. PMID- 16350040 TI - Safe, convenient pipetting station. AB - A simple convenient pipetting station is described that eliminates the need for mouth pipetting. Necessary components, fabrication procedures, and operating techniques are given. PMID- 16350042 TI - Method for collecting naturally occurring airborne bacterial spores for determining their thermal resistance. AB - [This corrects the article on p. 787 in vol. 30.]. PMID- 16350043 TI - Microwave sanitization of color additives used in cosmetics: feasibility study. AB - [This corrects the article on p. 206 in vol. 30.]. PMID- 16350041 TI - Microbial inhibition by an isolate of pediococcus from cucumber brines. AB - We reported earlier that Pediococcus cerevisiae FBB-61 inhibited Lactobacillus plantarum FBB-67 in mixed species inoculation used for the fermentation of brined cucumbers. Herein, 16 isolates of the Pediococcus genus from various sources were tested for inhibitory activity against L. plantarum and other microorganisms by a seeded-agar screening technique. Only two of the 16 isolates gave consistent and distinctive zones of inhibition, and both were isolated from fermenting cucumber brines on separate occasions. These two isolates did not inhibit each other but did inhibit the other 14 Pediococcus isolates in addition to L. plantarum. They also inhibited several other gram-positive bacteria, but not four species each of gram-negative bacteria and yeasts tested. Inoculation of cucumber juice broth with P. cerevisiae FBB-61 and L. plantarum WSO resulted in a drastic reduction in the plate count of L. plantarum WSO during day 1, but counts increased rapidly thereafter. Consequently, acid production by L. plantarum WSO was delayed. Noninhibitory isolates of Pediococcus had no appreciable effect on growth and acid production by L. plantarum WSO. PMID- 16350044 TI - Microbiology of ripening honey. AB - Two main groups of bacteria, classified as Gluconobacter and Lactobacillus, are present in ripening honey. A third bacterial group, classified as Zymomonas, and several types of yeast are occasionally isolated. Both in natural honey and in synthetic syrup the bacterial population decreases in the course of the ripening process. Lactobacillus and Gluconobacter disappear after minimum moisture (about 18%) is reached, but the former does so sooner than the latter. The presence of these bacteria in different parts of the bee has been also investigated. PMID- 16350045 TI - Semisolid fermentation of ryegrass straw. AB - Candida utilis, Aureobasidium pullulans, and Trichoderma viride were grown on pretreated ryegrass straw. The pretreatment consisted of hydrolysis of straw with 0.5 N H(2)SO(4) (water-substrate, 3:1) at 121 C, 100 C, and room temperature and adjustment of the hydrolysate to pH 4.5 to 5.0 with 5 N NH(4)OH. The 121 C pretreatment yielded a material containing 30% sugar and 2.3% N. The fermentation was carried on semisolid substrate (moisture level, 75%) in rotating jars for 2 to 3 days at room temperature. The organisms grew rapidly during the period from 18 to 42 h of incubation. During this period the number of microbial cells increased by 20- to 200-fold, and the level of NH(3)-N decreased from 1.3 to 0.9%. The fermentation resulted in a fourfold increase in protein, fivefold increase in crude fat, and 40% increase in the digestibility of straw. The best result in terms of increasing protein and digestibility of straw was obtained when C. utilis was grown on straw preheated at 121 C. PMID- 16350046 TI - Accelerated Fermentation of Brewer's Wort by Saccharomyces carlsbergensis. AB - A rapid procedure for wort fermentation with Saccharomyces carlsbergensis at 12 C is described. Fermentation time was reduced from 7 to 4 days with normal inoculum by shaking. Increasing the inoculation to 5 to 10 times normal and shaking resulted in complete fermentation in 3 days. Maximum yeast population was reached rapidly with the large inocula, but fermentation proceeded at approximately the same rate when inoculations in excess of four times the normal were used. Similar results were obtained with both small-scale (100 ml) and microbrew (2.4 liters) fermentations. PMID- 16350048 TI - Editorial Note. PMID- 16350047 TI - Microbial indicators of oil-rich salt marsh sediments. AB - Selected microbial parameters were monitored in sediments from a pristine and an oil-field salt marsh. Although numbers of hydrocarbonoclastic bacteria and fungi were significantly greater in the oil field, the values did not show a strong correlation with levels of hydrocarbons (r = 0.43 and r = 0.49, respectively). However, a high correlation was noted between ratios of hydrocarbonoclastic and total aerobic heterotrophic bacteria and levels of hydrocarbons as well as the relative concentration of hydrocarbons (ratio of hydrocarbons to chloroform extractables) (r = 0.87 and r = 0.77, respectively). Data suggest that this first ratio is a more valid microbial indicator of hydrocarbon abundance than other factors examined. Significant differences in the ratio of pigmented to total colony-forming units, the ratio of different to total colony-forming units, and the diversity index were noted between the natural and oil-field marsh. It is suggested that the presence of hydrocarbons alters the relative abundance of the most predominant aerobic heterotrophic bacteria. PMID- 16350049 TI - THE STREPTOCOCCI. PMID- 16350050 TI - RECENT CHEMICAL INVESTIGATIONS OF BACTERIAL TOXINS. PMID- 16350051 TI - SEROLOGICAL RELATIONS AMONG SPORE-FORMING ANAEROBIC BACTERIA. PMID- 16350052 TI - THE FIBRINOLYTIC ACTIVITY OF HEMOLYTIC STREPTOCOCCI. PMID- 16350053 TI - ACCESSORY GROWTH FACTORS FOR BACTERIA AND RELATED MICROORGANISMS. PMID- 16350054 TI - COLIFORM BACTERIA. PMID- 16350055 TI - QUANTITATIVE ABSOLUTE METHODS IN THE STUDY OF ANTIGEN-ANTIBODY REACTIONS. PMID- 16350056 TI - THE PATHOGENIC STAPHYLOCOCCI. PMID- 16350057 TI - THE EARLIER PHASES OF THE BACTERIAL CULTURE CYCLE. PMID- 16350058 TI - BACTERIAL DISSIMILATION OF CARBOHYDRATES. PMID- 16350059 TI - THE ADAPTIVE PRODUCTION OF ENZYMES BY BACTERIA. PMID- 16350060 TI - THE MICROBIOLOGY OF INSECTS: With Special Reference to the Biologic Relationships between Bacteria and Insects. PMID- 16350061 TI - THE MENINGOCOCCUS (NEISSERIA INTRACELLULARIS). PMID- 16350062 TI - NUTRITION OF THE DIPHTHERIA BACILLUS. PMID- 16350063 TI - ENZYMES CONCERNED IN THE PRIMARY UTILIZATION OF AMINO ACIDS BY BACTERIA. PMID- 16350064 TI - CORYNEBACTERIUM DIPHTHERIAE: A Correlation of Recorded Variations within the Species. PMID- 16350065 TI - NON-SPOREFORMING ANAEROBIC BACTERIA OF MEDICAL IMPORTANCE. PMID- 16350066 TI - THE CHEMISTRY AND SEROLOGY OF THE VIBRIOS. PMID- 16350067 TI - THE FILTRABLE MICROORGANISMS OF THE PLEUROPNEUMONIA GROUP. PMID- 16350069 TI - THE CHEMISTRY OF THE PROTEINS OF THE ACID-FAST BACILLI. PMID- 16350070 TI - THE YEASTS: Genetics, Cytology, Variation, Classification and Identification. PMID- 16350071 TI - THE CYTOLOGY OF BACTERIA. PMID- 16350072 TI - ANTAGONISTIC RELATIONS OF MICROORGANISMS. PMID- 16350073 TI - THE WASSERMANN ANTIGEN AND RELATED "ALCOHOL-SOLUBLE" ANTIGENS. PMID- 16350074 TI - THE FILTRABLE MICROORGANISMS OF THE PLEUROPNEUMONIA GROUP: (Appendix to Section on Classification and Nomenclature). PMID- 16350075 TI - FACTORS INFLUENCING BACTERIAL GROWTH IN BUTTER. PMID- 16350076 TI - THE GENUS SHIGELLA: (Dysentery Bacilli and Allied Species). PMID- 16350077 TI - THE PATHOGENIC RICKETTSIAE WITH PARTICULAR REFERENCE TO THEIR NATURE, BIOLOGIC PROPERTIES, AND CLASSIFICATION. PMID- 16350080 TI - IMMUNITY IN BRUCELLOSIS. PMID- 16350081 TI - ELEMENTARY BODIES OF VACCINIA. PMID- 16350082 TI - THE CYTOPHAGA GROUP: A CONTRIBUTION TO THE BIOLOGY OF MYXOBACTERIA. PMID- 16350083 TI - TISSUE PERMEABILITY AND THE SPREADING FACTORS IN INFECTION : A Contribution to the Host:Parasite Problem. PMID- 16350084 TI - THE TYPES MITIS, INTERMEDIUS AND GRAVIS OF CORYNEBACTERIUM DIPHTHERIAE: A Review of Observations during the Past Ten Years. PMID- 16350085 TI - THE SIGNIFICANCE OF THE V(I) ANTIGEN. PMID- 16350086 TI - STATISTICAL METHODS AND CONTROL IN BACTERIOLOGY. PMID- 16350088 TI - THE MODE OF ACTION OF SULFONAMIDES. PMID- 16350090 TI - THE CULTURE, GENERAL PHYSIOLOGY, MORPHOLOGY, AND CLASSIFICATION OF THE NON-SULFUR PURPLE AND BROWN BACTERIA. PMID- 16350087 TI - FACTORS INFLUENCING THE ENZYMIC ACTIVITIES OF BACTERIA. PMID- 16350092 TI - EARLY AMERICAN PUBLICATIONS RELATING TO BACTERIOLOGY: I. Textbooks, Monographs, Addresses, Etc. PMID- 16350093 TI - MICROBIOLOGICAL PROBLEMS IN THE PRESERVATION OF MEATS. PMID- 16350094 TI - THE PARASITIC ACTINOMYCETES AND OTHER FILAMENTOUS MICROORGANISMS OF THE MOUTH: A Review of Their Characteristics and Relationships, of the Bacteriology of Actinomycosis, and of Salivary Calculus in Man. PMID- 16350095 TI - RECENT ADVANCES IN OUR KNOWLEDGE OF THE PHYSIOLOGY OF MICROORGANISMS. PMID- 16350096 TI - THE SIGNIFICANCE OF THE BACTERIA AND THE PROTOZOA OF THE RUMEN OF THE BOVINE . PMID- 16350097 TI - THE OUTLINE CLASSIFICATION USED IN THE BERGEY MANUAL OF DETERMINATIVE BACTERIOLOGY. PMID- 16350098 TI - PHYSICAL METHODS OF STERILIZATION OF MICROORGANISMS. PMID- 16350100 TI - YEAST GENETICS : Life Cycles, Cytology, Hybridization, Vitamin Synthesis, and Adaptive Enzymes. PMID- 16350101 TI - ACTION OF MICROORGANISMS ON HYDROCARBONS. PMID- 16350103 TI - CHEMICAL NATURE AND BIOLOGICAL SPECIFICITY OF THE SUBSTANCE INDUCING TRANSFORMATION OF PNEUMOCOCCAL TYPES. PMID- 16350102 TI - AN ORIENTATION WITH RESPECT TO MEMBERS OF THE GENUS BACILLUS PATHOGENIC FOR INSECTS. PMID- 16350099 TI - RELATION OF BACTERIA TO VITAMINS AND OTHER GROWTH FACTORS. PMID- 16350104 TI - THE SIGNIFICANCE AND APPLICATIONS OF BACTERIOPHAGE IN BACTERIOLOGICAL AND VIRUS RESEARCH. PMID- 16350105 TI - RECENT ADVANCES IN BACTERIAL GENETICS. PMID- 16350106 TI - THE MECHANISM OF BIOLOGICAL NITROGEN FIXATION. PMID- 16350107 TI - USE OF MOVING AVERAGES AND INTERPOLATION TO ESTIMATE MEDIAN-EFFECTIVE DOSE: I. Fundamental Formulas, Estimation of Error, and Relation to Other Methods. PMID- 16350108 TI - BACTERIAL DISSOCIATION: A Critical Review of a Phenomenon of Bacterial Variation. PMID- 16350109 TI - MECHANISMS OF INFECTION AND IMMUNITY IN VIRUS DISEASES OF MAN. PMID- 16350110 TI - PROBLEMS OF AUTOTROPHY. PMID- 16350111 TI - SOME INTROSPECTIONS ON MOLD METABOLISM. PMID- 16350113 TI - THE YEAST-LIKE FUNGI: CANDIDA AND BRETTANOMYCES. PMID- 16350112 TI - THE THERMOPHILIC MICROORGANISMS. PMID- 16350114 TI - DYNAMICS AND MECHANISM OF IMMUNITY REACTIONS IN VIVO. PMID- 16350115 TI - THE INHIBITION OF REPRODUCTION OF PARASITES BY IMMUNE FACTORS. PMID- 16350116 TI - THE ENDOSPORE OF BACTERIA. PMID- 16350117 TI - CYTOCHEMICAL INTERPRETATION OF THE MECHANISM OF PENICILLIN ACTION. PMID- 16350118 TI - SURFACE ACTIVE AGENTS AND THEIR APPLICATION IN BACTERIOLOGY. PMID- 16350119 TI - MUCIN AS A RESISTANCE-LOWERING SUBSTANCE. PMID- 16350120 TI - CELLULAR STRUCTURES AND FUNCTIONS CONCERNED IN PARASITISM. PMID- 16350121 TI - PROPERTIES OF CERTAIN RAPIDLY ACTING BACTERIAL TOXINS AS ILLUSTRATED BY STREPTOLYSINS O AND S . PMID- 16350122 TI - CHARACTERISTICS OF PATHOGENIC SPIROCHETES AND SPIROCHETOSES WITH SPECIAL REFERENCE TO THE MECHANISMS OF HOST RESISTANCE. PMID- 16350123 TI - THE MOST ABUNDANT GROUPS OF BACTERIA IN SOIL. PMID- 16350124 TI - PROFESSOR HERBERT WILLIAM CONN AND THE FOUNDING OF THE SOCIETY. PMID- 16350125 TI - CORN STEEP LIQUOR IN MICROBIOLOGY. PMID- 16350126 TI - GROWTH REQUIREMENTS OF BACTERIAL VIRUSES. PMID- 16350127 TI - SOME CONSIDERATIONS OF THE BIOLOGICAL IMPORTANCE OF INTESTINAL MICROORGANISMS. PMID- 16350128 TI - THE RELATIONSHIP BETWEEN BACTERIA AND MYXOPHYCEAE. PMID- 16350129 TI - INFLUENCE OF NUTRITION IN EXPERIMENTAL INFECTION. PMID- 16350130 TI - IMMUNITY IN POLIOMYELITIS. PMID- 16350131 TI - THE "DELFT SCHOOL" AND THE RISE OF GENERAL MICROBIOLOGY. PMID- 16350132 TI - A MECHANICAL KEY FOR THE GENERIC IDENTIFICATION OF BACTERIA. PMID- 16350136 TI - SYMPOSIUM ON MICROBIAL TOXINS. PMID- 16350137 TI - SYMPOSIUM ON METHODOLOGY FOR SALMONELLA IN FOOD. PMID- 16350138 TI - SYMPOSIUM ON MICROBIOLOGY OF THE RUMEN. PMID- 16350139 TI - SYMPOSIUM ON THE MAINTENANCE OF CULTURES OF MICROORGANISMS. PMID- 16350140 TI - SYMPOSIUM ON METHODS FOR DETERMINING BACTERIAL CONTAMINATION ON SURFACES. PMID- 16350141 TI - SYMPOSIUM ON NEW ASPECTS OF WATER BACTERIOLOGY. PMID- 16350143 TI - SYMPOSIUM ON ANALYTICAL MICROBIOLOGY. PMID- 16350142 TI - SYMPOSIUM ON PETROLEUM MICROBIOLOGY. PMID- 16350144 TI - CELLULAR ORGANIZATION IN RELATION TO FUNCTION. PMID- 16350145 TI - SYMPOSIUM ON ENTEROBACTERIACEAE OF MEDICAL SIGNIFICANCE: ISOLATION, IDENTIFICATION, AND PATHOGENIC POTENTIALITIES. PMID- 16350146 TI - SYMPOSIUM ON PROBLEMS IN TAXONOMY. PMID- 16350147 TI - SYMPOSIUM ON INDUSTRIAL FERMENTATIONS. PMID- 16350148 TI - SYMPOSIUM ON BACTERIAL PIGMENTS. PMID- 16350149 TI - SYMPOSIUM ON MICROBIAL AMINO ACID METABOLISM. PMID- 16350150 TI - SYMPOSIUM ON PROBLEMS IN THE MICROBIOLOGICAL ANALYSIS OF FOODS. PMID- 16350151 TI - SYMPOSIUM ON VIRUS DIAGNOSIS IN THE PUBLIC HEALTH LABORATORY. PMID- 16350152 TI - SYMPOSIUM ON MICROBIAL AEROSOLS AND RESPIRATORY INFECTIONS. PMID- 16350153 TI - SYMPOSIUM ON ANTIMICROBIAL PRESERVATIVES. PMID- 16350154 TI - SYMPOSIUM ON THE ROLE OF CELL FRAGMENTS IN METABOLIC REACTIONS. PMID- 16350155 TI - SYMPOSIUM ON PREPARATION OF SAMPLES FOR MICROBIOLOGICAL ASSAY. PMID- 16350156 TI - SYMPOSIUM ON BACTERIAL SPORE GERMINATION. PMID- 16350157 TI - PSYCHROPHILIC BACTERIA. PMID- 16350158 TI - I. : Lysogenic Conversions in Bacteria. PMID- 16350159 TI - I. : Introduction. PMID- 16350160 TI - II. : Cytological Changes in Early Growth Phases. PMID- 16350161 TI - III. : Physiological Aspects of Growth Initiation. PMID- 16350162 TI - IV. : Dormancy, Germination, and Outgrowth. PMID- 16350164 TI - INTRODUCTION. PMID- 16350165 TI - II. THE IMPORTANCE OF ENVIRONMENTAL FACTORS IN THE INSECT-MICROBE ECOSYSTEM. PMID- 16350163 TI - VIRAL INTERFERENCE. SOME CONSIDERATIONS OF BASIC MECHANISMS AND THEIR POTENTIAL RELATIONSHIP TO HOST RESISTANCE. PMID- 16350166 TI - A REVIEW OF THE GENUS CANDIDA. PMID- 16350167 TI - PHOTOSYNTHETIC MECHANISMS IN BACTERIA AND PLANTS: DEVELOPMENT OF A UNITARY CONCEPT. PMID- 16350168 TI - CHLORAMPHENICOL. PMID- 16350170 TI - DISCUSSIONS OF PAPERS ON AIRBORNE ORGANISMS. PMID- 16350171 TI - DISCUSSIONS OF PAPERS ON RESPIRATORY SYSTEM AND AIRBORNE INFECTION. PMID- 16350169 TI - IMMUNOFLUORESCENT STAINING: THE FLUORESCENT ANTIBODY METHOD. PMID- 16350172 TI - AEROSOL INFECTION OF MAN WITH PASTEURELLA TULARENSIS. PMID- 16350173 TI - DISCUSSIONS OF PAPERS ON BACTERIAL DISEASES. PMID- 16350174 TI - DISCUSSIONS OF PAPERS ON VIRUS AND FUNGUS DISEASES. PMID- 16350176 TI - DISCUSSIONS OF PAPERS ON IMMUNOLOGY AND PUBLIC HEALTH. PMID- 16350175 TI - IMMUNOLOGICAL ASPECTS OF AIRBORNE INFECTION: SOME GENERAL CONSIDERATIONS OF RESPONSE TO INHALATION OF TOXINS. PMID- 16350177 TI - PART VII. CRITIQUE OF CONFERENCE. Chairman: Walsh McDermott. PMID- 16350178 TI - III. NITROGEN FIXATION BY ENZYME PREPARATIONS. PMID- 16350179 TI - THE FATTY ACIDS OF BACTERIA. PMID- 16350182 TI - DISCUSSION. PMID- 16350181 TI - GENETIC TRANSFORMATION OF RHIZOBIUM: A REVIEW OF THE WORK OF R. BALASSA. PMID- 16350180 TI - BACTERIOCINS AND BACTERIOCIN-LIKE SUBSTANCES. PMID- 16350183 TI - PHYTOTOXIC SUBSTANCES FROM SOIL MICROORGANISMS AND CROP RESIDUES. AB - [This corrects the article on p. 181 in vol. 28.]. PMID- 16350184 TI - DISCUSSION. PMID- 16350185 TI - Macrophage Function in Infectious Disease with Inbred Rabbits. PMID- 16350186 TI - Introductory remarks. PMID- 16350187 TI - Discussion. PMID- 16350189 TI - Discussion. PMID- 16350188 TI - Discussion. PMID- 16350190 TI - Discussion. PMID- 16350191 TI - Discussion. PMID- 16350192 TI - Discussion. PMID- 16350193 TI - Author's Comments on the Discussion. PMID- 16350194 TI - Discussion. PMID- 16350195 TI - Discussion. PMID- 16350196 TI - Discussion. PMID- 16350197 TI - Discussion. PMID- 16350198 TI - Discussion. PMID- 16350199 TI - Discussion. PMID- 16350200 TI - Discussion. PMID- 16350201 TI - Discussion. PMID- 16350202 TI - Comparative ecology of respiratory mycotic disease agents. PMID- 16350203 TI - Epidemiological aspects of venezuelan equine encephalitis virus infections. PMID- 16350205 TI - Epidemiology of airborne staphylococcal infections. AB - [This corrects the article on p. 660 in vol. 30.]. PMID- 16350204 TI - Various molecular species of interferon induced by viral and nonviral agents. PMID- 16350206 TI - Arrangement of base sequences in deoxyribonucleic Acid. PMID- 16350207 TI - Deoxyribonucleic Acid of the blue-green algae (cyanophyta). PMID- 16350208 TI - Extracellular lipids of yeasts. AB - [This corrects the article on p. 194 in vol. 31.]. PMID- 16350209 TI - Genetic Analysis and Genome Structure in Streptomyces coelicolor. AB - [This corrects the article on p. 388 in vol. 31.]. PMID- 16350210 TI - Intergeneric bacterial matings. PMID- 16350211 TI - Editorial. PMID- 16350212 TI - Letters to the editor. PMID- 16350213 TI - Letters to the editor. PMID- 16350214 TI - Antibiotic inhibitors of the bacterial ribosome. AB - [This corrects the article on p. 493 in vol. 32.]. PMID- 16350215 TI - Thermophilic blue-green algae and the thermal environment. AB - [This corrects the article on p. 485 in vol. 33.]. PMID- 16350217 TI - Survey of numerical techniques for grouping. AB - [This corrects the article on p. 379 in vol. 35.][This corrects the article on p. 381 in vol. 35.][This corrects the article on p. 384 in vol. 35.][This corrects the article on p. 387 in vol. 35.][This corrects the article on p. 388 in vol. 35.][This corrects the article on p. 389 in vol. 35.]. PMID- 16350218 TI - Molecular structure of bacterial plasmids. AB - [This corrects the article on p. 369 in vol. 36.]. PMID- 16350219 TI - Mechanisms of virus pathogenicity. AB - [This corrects the article on p. 302 in vol. 36.]. PMID- 16350220 TI - Peptidoglycan types of bacterial cell walls and their taxonomic implications. AB - [This corrects the article on p. 409 in vol. 36.]. PMID- 16350221 TI - Chemistry and biology of the polyene macrolide antibiotics. AB - [This corrects the article on p. 166 in vol. 37, PMID: 4578757.]. PMID- 16350222 TI - Aldolase of lactic Acid bacteria: a case history in the use of an enzyme as an evolutionary marker. AB - [This corrects the article on p. 453 in vol. 37.]. PMID- 16350224 TI - The Hemolysins of Staphylococcus aureus. AB - [This corrects the article on p. 321 in vol. 39.]. PMID- 16350223 TI - Bacteriophages of Bacillus subtilis. AB - [This corrects the article on p. 294 in vol. 39.][This corrects the article on p. 295 in vol. 39.][This corrects the article on p. 298 in vol. 39.]. PMID- 16350225 TI - Recalibrated Linkage Map of Escherichia coli K-12. AB - [This corrects the article on p. 116 in vol. 40.]. PMID- 16350216 TI - Bacterial ribosome. PMID- 16350226 TI - Uniform nomenclature for bacterial plasmids: a proposal. AB - [This corrects the article on p. 168 in vol. 40.]. PMID- 16350228 TI - Energy conservation in chemotrophic anaerobic bacteria. AB - [This corrects the article on p. 100 in vol. 41.]. PMID- 16350229 TI - Mycobacterium. AB - [This corrects the article on p. 217 in vol. 41.]. PMID- 16350227 TI - Degradation of purines and pyrimidines by microorganisms. AB - [This corrects the article on p. 403 in vol. 40.]. PMID- 16350230 TI - Role of ribonucleic Acid polymerase in gene selection in procaryotes. AB - [This corrects the article on p. 568 in vol. 41.]. PMID- 16350231 TI - An editorial. PMID- 16350232 TI - The biology of methanogenic bacteria. AB - [This corrects the article on p. 517 in vol. 41.]. PMID- 16350234 TI - Biosynthesis of cell walls of fungi. AB - [This corrects the article on p. 134 in vol. 43.]. PMID- 16350233 TI - Algal toxins. PMID- 16350235 TI - Nonenteric Toxins of Staphylococcus aureus. AB - [This corrects the article on p. 337 in vol. 43.]. PMID- 16350236 TI - Evaluation of nitrogen fixation by bacteria in association with roots of tropical grasses. AB - [This corrects the article on p. 494 in vol. 44.][This corrects the article on p. 496 in vol. 44.][This corrects the article on p. 498 in vol. 44.][This corrects the article on p. 499 in vol. 44.][This corrects the article on p. 503 in vol. 44.][This corrects the article on p. 506 in vol. 44.][This corrects the article on p. 507 in vol. 44.][This corrects the article on p. 512 in vol. 44.][This corrects the article on p. 513 in vol. 44.][This corrects the article on p. 517 in vol. 44.]. PMID- 16350237 TI - Distribution of isoprenoid quinone structural types in bacteria and their taxonomic implications. AB - [This corrects the article on p. 318 in vol. 45.]. PMID- 16350238 TI - Methane-oxidizing microorganisms. AB - [This corrects the article on p. 586 in vol. 45.]. PMID- 16350239 TI - Linkage Map of Escherichia coli K-12, Edition 7. AB - [This corrects the article on p. 182 in vol. 47.]. PMID- 16350240 TI - Energy conservation in acidophilic bacteria. AB - [This corrects the article on p. 580 in vol. 47.][This corrects the article on p. 584 in vol. 47.][This corrects the article on p. 594 in vol. 47.]. PMID- 16350242 TI - Biosynthesis and metabolism of arginine in bacteria. AB - [This corrects the article on p. 351 in vol. 50.]. PMID- 16350244 TI - Mechanism of bactericidal action of aminoglycosides. AB - [This corrects the article on p. 341 in vol. 51.]. PMID- 16350246 TI - Biophysics of bacterial walls viewed as stress-bearing fabric. AB - [This corrects the article on p. 337 in vol. 52.]. PMID- 16350247 TI - Glutamine Metabolism and Cycling in Neurospora crassa. AB - [This corrects the article on p. toc in vol. 54.]. PMID- 16350248 TI - Linkage Map of Escherichia coli K-12, Edition 8. AB - [This corrects the article on p. 131 in vol. 54.]. PMID- 16350249 TI - In memorium. PMID- 16350251 TI - Listeria monocytogenes, a Food-Borne Pathogen. AB - [This corrects the article on p. 485 in vol. 55.]. PMID- 16350250 TI - Transformation in fungi. AB - [This corrects the article on p. 159 in vol. 53.]. PMID- 16350252 TI - Bacteriophage lambda as a cloning vector. AB - [This corrects the article on p. 582 in vol. 56.]. PMID- 16350254 TI - The alphaviruses: gene expression, replication, and evolution. AB - [This corrects the article on p. 496 in vol. 58.]. PMID- 16350253 TI - Polypeptides of Treponema pallidum: Progress toward Understanding Their Structural, Functional, and Immunologic Roles. AB - [This corrects the article on p. 760 in vol. 57.]. PMID- 16350256 TI - 6 major risk factors for an older heart. Countering just one or two, now, could turn your cardiac clock back. PMID- 16350257 TI - Surprise finding raises questions about heart attack care. PMID- 16350258 TI - Chocolate, tea: health foods? How plant chemicals may help foster cardiovascular health. PMID- 16350259 TI - Cardiac health while you sleep. An A-to-Zzzz guide to restful nights and more zestful days. PMID- 16350260 TI - Heart device recalls: your move. Questions and answers in the wake of malfunction reports. PMID- 16350261 TI - I had bypass surgery in early 2004, and a couple of months later began to notice chest pain on exertion, even just taking a shower. Then at Thanksgiving last year, I found that I was able to do more, especially walking. But I still have daily chest pain. Should I be thinking about another bypass or angioplasty, or just stick with the medications? I'm 55. PMID- 16350262 TI - A how-to for a healthier you. A new online tool flags your health risks. PMID- 16350263 TI - A heart remembers. Another reason tight control matters. PMID- 16350264 TI - Lows woes. Understanding severe low blood glucose. PMID- 16350265 TI - Sailing on a wave of new research. Part 2. PMID- 16350266 TI - Your holiday feast. Make-ahead masterpiece. Say good-bye to last-minute hassles with a menu you can prepare in advance. PMID- 16350267 TI - Your holiday feast. Traditional fare with a twist. Shake up your holiday meal with an Asian-inspired menu. PMID- 16350268 TI - Your holiday feast. Crowd-pleasing cooking. When your guest list is long, this menu helps you keep things simple. PMID- 16350269 TI - Keep it clean. How to avoid a scrape with infection. PMID- 16350270 TI - Research profile. Weak bones & type 1. What's the connection? Mari Palta, Ph.D. PMID- 16350271 TI - Evaluation of farm management practices as risk factors for clinical listeriosis and fecal shedding of Listeria monocytogenes in ruminants. AB - OBJECTIVE: To assess seasonal variation in prevalence of Listeria monocytogenes on ruminant farms and identify management practices associated with ruminant listeriosis and fecal shedding of L. monocytogenes. STUDY DESIGN: Case-control study. SAMPLE POPULATION: 2056 samples of feces, feed, soil, and water from 24 case farms with listeriosis and 28 control farms without listeriosis. PROCEDURE: Samples were collected and evaluated via bacterial culture for L. monocytogenes. Univariate associations between farm management practices and listeriosis and fecal shedding of L. monocytogenes were assessed. Multivariate models were developed to identify farm management practices associated with listeriosis and fecal shedding of L. monocytogenes. RESULTS: The prevalence of L. monocytogenes on cattle, goat, and sheep farms was seasonal, especially in fecal samples, with peak prevalence in winter. Although the prevalence of L. monocytogenes in feedstuffs from small-ruminant farms also peaked during winter, the bacterium was detected at a constant rate in cattle farm feedstuffs throughout the year. Farm management practices, animal health and hygiene, and feedstuff quality and storage were associated with ruminant listeriosis and fecal shedding of L. monocytogenes. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the prevalence of L. monocytogenes on ruminant farms is seasonal, management practices are associated with ruminant listeriosis and fecal shedding of L. monocytogenes, and the epidemiologic features of listeriosis differ in cattle versus small ruminants. Awareness of risk factors may be used to develop control measures to reduce animal disease and introduction of L. monocytogenes into the human food chain. PMID- 16350272 TI - Thalidomide therapy and deep venous thrombosis in multiple myeloma. PMID- 16350273 TI - 115 kHz tuning repetition rate ultrahigh-speed wavelength-swept semiconductor laser. AB - We demonstrate an ultrahigh-speed wavelength-swept semiconductor laser using a polygon-based wavelength scanning filter. With a polygon rotational speed of 900 revolutions per second, a continuous wavelength tuning rate of 9200 nm/ms and a tuning repetition rate of 115 kHz were achieved. The wavelength tuning range of the laser was 80 nm centered at 1325 nm, and the average polarized output power was 23 mW. PMID- 16350275 TI - Extending the array of crosslinkers suitable for the preparation of polymethacrylate-based monoliths. AB - The in situ preparation of monolithic capillary columns comprising copolymers of butyl methacrylate with ethylene dimethacrylate, diethylene glycol dimethacrylate, triethylene glycol dimethacrylate, and pentaerythritol tetraacrylate using thermal polymerization within 250 microm ID capillaries and their application for micro-HPLC separations of proteins has been studied. For all crosslinkers, optimization of the porogenic mixture consisting of 1-propanol and 1,4-butanediol yielded monoliths with pore sizes above 1 microm suitable for rapid separations at low back pressure. Very good separations were achieved for a protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin with all tested columns. PMID- 16350274 TI - Quantitative spectroscopy of superficial turbid media. AB - We report a novel diffuse optical spectroscopy probe design for determining optical properties of superficial volumes of turbid samples. The fiber-based probe employs a highly scattering layer placed in contact with the sample of interest. This layer diffuses photons from a collimated light source before they enter the sample and provides a basis for describing light transported in superficial media by the diffusion approximation. We compare the performance of this modified two-layer diffusion model with Monte Carlo simulations. A set of experiments that demonstrate the feasibility of this method in turbid tissue phantoms is also presented. Optical properties deduced by this approach are in good agreement with those derived by use of a benchmark method for determining optical properties. The average interrogation depth of the probe design investigated here is estimated to be less than 1 mm. PMID- 16350276 TI - Integration of genomic data in Electronic Health Records--opportunities and dilemmas. AB - OBJECTIVES: In this paper we give an overview about the challenge the postgenomic era poses on biomedical informaticists. The occurrence of new (genomic) data types necessitates new data models, new viewing metaphors and methods to deal with the disclosure of genomic data. We discuss integration issues when inferring phenotype and genotype data. Another challenge is to find the right phenotype to genotype data in order to get appropriate case numbers for sound clinical genotype-phenotype inference studies. METHODS: Genomic data could be integrated in an Electronic Health Record (EHR) in several ways. We describe patient centered and pointer-based integration strategies and the corresponding data types and data models. The inference mechanisms for the interpretation of row data contain different agents. We describe vertical, horizontal and temporal agents. RESULTS: We have to deal with several new data types, not being standardized for EHR integration. Genomic data tends to be more structured than phenotype data. Beyond the development of new data models, vertical, horizontal and temporal agents have to be developed in order to link genotype and phenotype. As the genomic EHR will contain very sensitive data, confidentiality and privacy concerns have to be addressed. CONCLUSIONS: Given the necessity to capture both environment and genomic state of a patient and their interaction, clinical information systems have to be redesigned. While genotyping seems to be automatable easily, this is not the case for clinical information. More integration work on terminologies and ontologies has to be done. PMID- 16350277 TI - [Highlights from the 77th Annual Meeting of the American Heart Association, November 2004, New Orleans, Louisiana, USA]. PMID- 16350278 TI - [Abstracts of the 29th Scientific Congress of the German League Against High Blood Pressure and the German Society of Hypertension. Berlin, 23-25 November 2005]. PMID- 16350279 TI - Abstracts of the 46th Annual Conference of the Indian Society of Gastroenterology in association with the Indian Association for the Study of the Liver and the Society of Gastrointestinal Endoscopy of India, Visakhapatnam, India, November 11 15, 2005. PMID- 16350280 TI - Proceedings of the International Symposium Nitric Oxide, Cytokines and Inflammation, Rio de Janeiro, Brazil, June 6-9, 2004. PMID- 16350282 TI - Circumcision prevents HIV infection. PMID- 16350281 TI - Food for thought. PMID- 16350283 TI - Proceedings of the Second Biennial Conference: Craniofacial Skeletal Bioengineering. Pacific Grove, CA, USA, 27-30 August 2004. PMID- 16350284 TI - Disabling disability claims in gastroenterology. Interview by Paul Adams. PMID- 16350294 TI - The pamphlet collection at NLM. PMID- 16350295 TI - [Role of the Committee for Postgraduate Training in the new system of specialization for psychiatry in Japan]. PMID- 16350296 TI - A short history of the Health Sciences Historical Collections (HSHC), University of Tennessee Health Sciences Library, Memphis. PMID- 16350297 TI - The correspondence of the Tring Museum at the Natural History Museum, London. PMID- 16350298 TI - The Cyril Dean Darlington Papers. PMID- 16350299 TI - Joseph Henry Woodger (1894-1981) Papers at the University College London. PMID- 16350300 TI - Where to look next? Agricultural archives as resources for the history of genetics. PMID- 16350301 TI - A cultural history of heredity. PMID- 16350302 TI - Shaping the profession: James Law and the rise of professionalism in the U.S. PMID- 16350303 TI - Dr. Benjamin Rush, a true American hero. PMID- 16350304 TI - Forgotten veterinarians, 4. John Boyd Dunlop and his pneumatic tire. PMID- 16350305 TI - Animal therapy over the ages: 14. amulets and transfer charms. PMID- 16350306 TI - History of the surgical laboratory. PMID- 16350307 TI - [Present status and subjects in the new system of postgraduate clinical training as viewed by the Committee for Postgraduate Clinical Training in Psychiatry in Japan]. PMID- 16350309 TI - [New system for postgraduate psychiatry education as viewed from a psychiatric hospital in Japan]. PMID- 16350308 TI - [Mandatory postgraduate training in psychiatry in Yokohama City University and the affiliated medical facilities under the new system of postgraduate clinical education in Japan]. PMID- 16350311 TI - [Ideal postgraduate psychiatry education as viewed by a recipient of the education]. PMID- 16350310 TI - [Present status and problems in the new system of postgraduate psychiatry training in the psychiatry department of Nagasaki University Hospital]. PMID- 16350312 TI - [Psychiatric care as viewed by young psychiatrists]. PMID- 16350313 TI - [Present status and problems in psychiatric care in cooperation with emergency medical services at the general hospital without psychiatric ward]. PMID- 16350314 TI - [Community psychiatric care as viewed by young psychiatrists in Japan]. PMID- 16350315 TI - [Comparison of emergency psychiatric services in Tokyo and Chiba prefecture]. PMID- 16350316 TI - [Contribution of the Japan Young Psychiatrists Organization to international activities of psychiatrists]. PMID- 16350317 TI - [Basic clinical training in psychiatry]. PMID- 16350318 TI - [Psychologic interviews for detecting psychiatric symptoms in patients with mental disorders]. PMID- 16350319 TI - Treatment of obese female and male SHHF/Mcc-fa(cp) rats with antihypertensive drugs, nifedipine and enalapril: effects on body weight, fat distribution, insulin resistance and systolic pressure. AB - Little is known about the effects of common antihypertensive drugs in obese, insulin-resistant females. Nine-month-old obese female SHHF/Mcc-fa(cp) rats that received either nifedipine, a calcium channel antagonist, or enalapril, an angiotensin-converting-enzyme inhibitor, for three months were compared with untreated SHHF/Mcc-fa(cp) rats (controls). After one month, nifedipine significantly decreased body weight in obese females compared to either enalapril or controls. After three months of treatment, total, abdominal, and subcutaneous fat masses were decreased in obese females given nifedipine compared to either enalapril or controls. Enalapril treatment was associated with a redistribution of fat mass from abdominal to subcutaneous depots. Nifedipine reduced plasma triglyceride and fasting glucose levels and improved insulin response to an oral glucose load in obese females, whereas enalapril did not appear to affect glycemic control. Systolic pressure was not significantly decreased until after two months of treatment with nifedipine or three months of treatment with enalapril in obese females and may have coincided with improvement in insulin resistance. Similarly, plasma atrial natriuretic peptide concentrations were significantly lower in obese females given nifedipine. To determine how obese males responded to a calcium channel antagonist, six-month-old obese male SHHF/Mcc-fa(cp) rats were treated for three months with either nifedipine or placebo (controls). Nifedipine-treated obese males showed a mild but significant decrease in weight gain that was due to a decrease in fat deposition in both subcutaneous and abdominal depots and systolic blood pressure was significantly reduced after one month of treatment. Nifedipine did not affect other plasma biochemical parameters in obese males. In conclusion, nifedipine improved systolic pressure and glycemic control in obese female SHHF/Mcc-fa(cp) rats, effects that may be associated with a marked loss in body weight and fat mass and improved lipid metabolism. Nifedipine-treated obese males exhibited only a diminished weight gain that was not associated with changes in diabetic characteristics. PMID- 16350320 TI - Metabolism of different adipose tissues in vivo in the rat. AB - To explore regional differences in triglyceride retention in white adipose tissues of growing male rats, the mass of adipocytes from epididymal, retroperitoneal, inguinal, and mesenteric tissues were followed with time. In order to attempt to explain regional differences, adipose tissue metabolism was studied in vivo and in vitro. (U-14C) oleic acid in sesame oil was given by gastric gavage to conscious male and female rats, and accumulation and half-life of radioactivity measured. Lipoprotein lipase activity and lipolysis were studied in vitro. Adipocyte triglyceride mass increased linearly in all the depots during 4 months of observation.The increase in mass was more pronounced in retroperitoneal (0.31 microg) and epididymal (0.30 microg) than in mesenteric (0.11 microg) or inguinal (0.05 microg) adipocytes. In the fed state label from (U-14C) oleic acid first increased with time in liver, muscle, and adipose tissues. In the liver radioactivity peaked at 4 hours, and was not measurable in either liver or muscle after a time point between 24 hours to 1 week. In contrast label continued to increase in adipose tissues up to about 16 hours to 24 hours, suggesting transfer of label by recirculation from liver and muscle to adipose tissues. Thereafter the radioactivity decreased. When expressed per adipocyte uptake of label was not significantly different between white adipose tissues. The rate of decrease between 7 days and 4 months was, however, more rapid in mesenteric and inguinal than, particularly, epididymal, and, probably, retroperitoneal adipocytes. These results were partly parallel to in vitro data on lipoprotein lipase activity, which was not different between depots, and the rate of lipolysis, which was higher in mesenteric than other adipocytes. These results suggest that differences in weight increase of adipose tissue regions are due mainly to differences in the rate of mobilization of adipocyte triglycerides. When expressed per gram triglyceride, uptake and mobilization of label were clearly more rapid in mesenteric than other white adipose tissues. This is probably explained by a combination of a higher adipocyte density plus the metabolic characteristics of adipocytes in this depot. Since mesenteric adipose tissue is smaller than the other depots studied, the absolute contribution of this tissue to the energy supply of the body is probably not different from that of other adipose tissues, however. A large uptake and short half life was observed in interscapular adipose tissue. This region contains brown adipocytes, and the results therefore suggest that lipid uptake for thermogenic purposes is of a considerable magnitude. It was concluded that among white adipose tissues, the mesenteric tissue has a rapid turnover of triglyceride. This is probably due to a combination of a high density and specific metabolic characteristics of these adipocytes. Factors in the microenvironment of adipocytes probably contribute to the high turnover either directly, or by modification of cellular characteristics. PMID- 16350321 TI - Serum gamma-glutamyl transpeptidase levels and hypertension in non-drinkers: a possible role of fatty liver in the pathogenesis of obesity related hypertension. AB - The relationships between increases in body mass index (BMI) and increases in hypertension were compared between non-drinkers with elevated serum gamma glutamyl transpeptidase (gamma-GTP) levels (> or = 50 U/l) and those with normal levels, who comprised 10,952 men and 22,107 women aged 40-59 years recruited from an occupational health clinic. Hypertension was found in 16.1% and 13.5% of the men and women, and elevated serum g-GTP was found in 10.8% and 2.8% of the men and women, respectively. The prevalences of hypertension and elevated serum gamma GTP levels were both increased with increased BMI. Hypertension was, however, shown to be 1.5 times more prevalent in the persons with elevated serum gamma-GTP levels than in those with normal levels in both sexes, even after adjusting for BMI by a multiple logistic analysis. It can be concluded that elevations of serum gamma-GTP, which are probably a reflection of fatty liver in the non-drinkers, are closely related to the development of hypertension associated with increased obesity. PMID- 16350322 TI - Utility of a C-jun microsatellite marker in determining gene dosage for fatty (fa). AB - The Zucker fatty (fa) mutation provides a genetic model for obesity and non insulin dependent diabetes mellitus. The molecular pathogenesis of the metabolic phenotype of these animals is not known. Detailed molecular maps of the region surrounding the fa locus on rat chromosome 5 can be used for positional cloning experiments as well as to permit genotyping of animals from appropriate crosses before the confounding metabolic effects of obesity have occurred. We describe the development of a polymerase chain reaction (PCR) assay for a polymorphic simple sequence repeat (SSR) in the promoter region of the protooncogene c-Jun. This assay was used to position c-Jun 4.5cM proximal to the fa locus in 111 F2 progeny of a 13MBN fa/+ F1 intercross. Concurrent use of the c-Jun SSR with a previously described assay for a microsatellite in the glucose transporter, Glut1, permits rapid and accurate assessment of genotypes at the fa locus in animals of any age using minimal amounts of DNA. A strategy is described which minimizes the error rate in assigning genotype at the fatty locus for backcross and intercross progeny. PMID- 16350323 TI - Commentary on classics in obesity. 6. Science and politics of hunger. PMID- 16350324 TI - An explanation of hunger. 1911. PMID- 16350325 TI - Contributions to the physiology of the stomach.--II. The relation between the contractions of the empty stomach and the sensation of hunger. 1912. PMID- 16350326 TI - A discussion of hazardous child labor. PMID- 16350327 TI - Impact of bioterrorism preparedness on the public health response to the 2003 2004 influenza vaccine crisis. PMID- 16350328 TI - Injuries in child laborers in the informal sector in Mexico City, Mexico, 1997. PMID- 16350329 TI - Green tobacco sickness in children and adolescents. AB - Tobacco (Nicotiana tabacum) is cultivated in more than 100 countries, and in 2004, some 5.73 million metric tons dry weight of tobacco were grown worldwide. The top five tobacco producers forecast for 2004 are China (2.01 million metric tons; 35.1%), Brazil (757 thousand metric tons; 13.2%), India (598 thousand metric tons; 10.4%), United States (358 thousand metric tons; 6.2%), and Malawi (138 thousand metric tons; 2.4%). Together, these five countries account for two thirds of worldwide tobacco production. Tobacco farming presents several hazards to those who cultivate and harvest the plant. Although some of these hazards, such as pesticide exposure and musculoskeletal trauma, are faced by workers in other types of agricultural production, tobacco production presents some unique hazards, most notably acute nicotine poisoning, a condition also known as green tobacco sickness (GTS). GTS is an occupational poisoning that can affect workers who cultivate and harvest tobacco. It occurs when workers absorb nicotine through the skin as they come into contact with leaves of the mature tobacco plant. GTS is characterized largely by nausea, vomiting, headache, muscle weakness, and dizziness. Historically, children have played a role in agricultural production in the United States, and they continue to do so today. This includes tobacco farming. The North American Guidelines for Children's Agricultural Tasks, a set of injury prevention guidelines prepared by the National Children's Center for Rural and Agricultural Health and Safety, lists GTS as one of several hazards children face when working on tobacco farms. Children 17 years of age and younger who work on U.S. tobacco farms come from three main groups: members of farm families, migrant youth laborers (primarily Latinos), and other hired local children. All three groups are at risk for GTS. Beyond the U.S., tobacco production using child labor is an emerging topic of concern in developing nations. An international movement, advocated by the Eliminating Child Labor in Tobacco Foundation, is underway to restrict child labor in tobacco production. GTS is a unique occupational poisoning associated with tobacco farming. We suspect that many public health practitioners, clinicians, advocates, and researchers are unaware of GTS among children and adolescents. In this article, we provide an overview of GTS among young people who work in tobacco, summarize reports documenting pediatric GTS cases, explain GTS etiology, and present three case studies of pediatric GTS in Kentucky. In addition, we discuss the need for expanded surveillance and prevention of GTS, both in the United States and globally. PMID- 16350330 TI - Hazardous child labor: lead and neurocognitive development. AB - Hazardous child labor is challenging to define and quantify in the context of acute or chronic toxic exposures--either of which may cause significant disease and disability. Epidemiologic occupational studies in adults have documented many harmful outcomes secondary to exposure to toxic substances. Occupational surveillance efforts often have focused on acute injuries because they are more readily identified. Fassa has been able to compile data concerning injuries to child laborers but notes, "There is great need for studies in developing countries ... on the impact of child labour on illness." Although injuries may be underreported or undocumented among child laborers, acute injury is, at least, potentially recordable. This is in sharp contrast to toxic exposures, where exposure assessment is usually difficult and costly. This is especially true when it is done after an exposure has taken place. The outcomes of most toxic workplace exposures to children remain unknown. The Agency for Toxic Substances and Disease Registries (ATSDR) notes that in similar environments, children may have greater exposures than adults: "Pound for pound of body weight, children drink more water, eat more food, and breathe more air than adults," and "In some instances, children are less able than adults to detoxify chemicals and are thus more vulnerable." Children who begin work at an early age have many more years to develop illness than an adult doing the same work. A household survey of child laborers in Ethiopia found that a high proportion (greater than 90%) of children in both urban and rural areas of the country reported non-use of protective equipment. However, it is the experience of present author D.P. that this equipment is rarely if ever adequate, even when made available. For example, protective equipment such as respirators or impermeable gloves are designed for adults, and thus do not properly fit children. In defining hazardous child labor, the larger context of public health cannot be ignored. The study of child labor needs to take into account the baseline health of exposed individuals. In developing countries, the poorest and most vulnerable children are most often involved in work in order to earn money for survival. These children are also likely to already lack basic necessities of food and medical care, predisposing them to diarrhea, anemia, and micronutrient deficiencies. Underlying nutritional conditions may make children more susceptible to the effects of toxic substances such as lead. In addition, child laborers also may be exposed to lead and other toxic substances from their poor living conditions. PMID- 16350331 TI - Challenges in determining how child work affects child health. AB - Credible findings from well-crafted research studies are essential in assessing the impact of child work on children's health. Researchers, however, encounter significant challenges in defining the relevant group of workers for a study and identifying an appropriate comparison group. This article describes some of those challenges and explains how choices about study and comparison groups can lead to biased research results. When selecting study groups, researchers should be aware that the impact of work on health may depend on the type and intensity of the work, and on the context in which it occurs. They should avoid drawing conclusions about the health effects of particular work situations from studies of very heterogeneous groups of workers and should not overgeneralize from studies of more homogenous groups. When choosing comparison groups, researchers should select children whose health outcomes are likely to be comparable to the outcomes working children would experience if they did not work. In particular, researchers should attempt to find children who are similar to the workers of interest on relevant non-work characteristics, including socioeconomic status and levels of parental education. In addition, they should consider the extent to which healthier children are more likely to select into the labor force as a result of decisions by parents or employers, or due to their own greater fitness. Ideally, studies of the health effects of child work should use multiple comparison groups, including children who work in relatively safe, non-strenuous occupations. PMID- 16350333 TI - The SIMPOC Philippine Survey of Children 2001: a data source for analyzing occupational injuries to children. AB - Child labor is a global issue that exists in both industrialized and developing countries. With the unanimous adoption of International Labour Organization (ILO) Convention No. 182 in 1999 calling for the immediate elimination of the worst forms of child labor, ratifying member countries have committed themselves to identifying hazardous work for children in the context of their respective legislative frameworks. Part of tackling the problem of child labor is knowing what types of occupational activities children are engaged in, what types of work environments they are under, and what risks of injuries and illnesses they are exposed to while working. Using the Philippines as a country example, this study introduces a promising data source on children's work and presents a methodology for examining hazardous work to children through the examination of injury rates. Data for this study rely primarily on a nationally representative dataset from the Survey of Children 2001 carried out by the Philippine National Statistics Office, covering the months of October 2001 to September 2002. PMID- 16350334 TI - Child labor, gender, and health. AB - It is often forgotten that child labor is part of a multi-generational problem due in part to the failure to educate girls. Although the literacy rate for women has improved over the last two decades, in many countries it is less than half that of their male counterparts. This in turn leads to nutritional deficiencies, poverty, and poor health. While many researchers address the immediate health effects of child labor on the child laborers, this article addresses the issue of child labor from a broader perspective, one that identifies child labor as a contributor to intergenerational poverty, malnutrition, and limited educational attainment. Child labor and nutrition are important issues in both educational attainment and health status. PMID- 16350332 TI - Nontraditional work factors in farmworker adolescent populations: implications for health research and interventions. AB - Agriculture has been documented to be one of the most hazardous work environments for both adults and children. Adolescents may be especially vulnerable to adverse health effects from agricultural exposures due to the rapid growth and development experienced during those years. Separating the occupational, economic, and social issues in this population is difficult. Weak regulatory protection, lack of compliance with existing regulations, and gaps in service provision characterize the working conditions of adolescent farmworkers. Although there is increasing research on the impact of work organization on mental and physical health in adult working populations, there is a scarcity of research focused on this concept in young workers--and it remains virtually unaddressed in young farmworkers. Work characteristics of the informal work sector, better delineated in international literature, should be considered when planning research or interventions in this at-risk population. Further, the population of adolescent farmworkers is diverse, and research strategies and interventions need to be targeted and tailored to the heterogeneous groups. This article addresses some of the nontraditional work factors associated with the less-than-formal work organization and environments in the farmworker adolescent population and how these factors may inform the planning of research and interventions. Specifically, mobility, cultural patterns and social networks, alternative sampling strategies, alternative delivery of health care and education, and involvement of a wide range of players in the work environment of adolescent farmworkers should all be considered when conducting research or planning programs for this population. PMID- 16350335 TI - Child labor in Ghana cocoa production: focus upon agricultural tasks, ergonomic exposures, and associated injuries and illnesses. AB - OBJECTIVES: The goal of this study was to determine the occupational hazards experienced by children harvesting cocoa in western Ghana in order to design a vocational literacy life skills curriculum and radio social messaging campaign with a safety component to decrease hazardous work exposures in child agricultural work. METHODS: An observational analysis was conducted of children aged 9 through 17 based upon personal interviews of agricultural workers, focus groups, and direct observation of work practices and activities. Job site analysis incorporated task mapping, job hazard review, and a review of equipment and use of protective gear. RESULTS: Children and young people aged 9 through 17 are exposed to hazardous occupational exposures including strenuous work, sharp tools, and pesticides. Lack of training in proper safety practices and inadequate personal protective equipment were commonly noted. Injuries and illnesses included musculoskeletal disorders, sprains, strains, lacerations to the head, fractures, eye injuries, rashes, and coughing. CONCLUSION: Children working in cocoa harvesting are exposed to physical and chemical hazards without proper training or personal protective equipment. Unless safety interventions occur, there are potential long-term adverse health consequences. PMID- 16350337 TI - Child labor and musculoskeletal disorders: the Pelotas (Brazil) epidemiological survey. AB - OBJECTIVES: [corrected] This article describes the prevalence of musculoskeletal pain in several anatomic sites in children and teens, and investigates, while adjusting for potential confounders, the association between musculoskeletal pain and back pain and the following: age, gender, sports practice, use of computer/video games/television, school attendance, intensity of involvement in household domestic activities, care of other children, care of sick/elderly family members, work activities, and workloads. METHODS: We conducted a cross sectional study interviewing 3,269 children aged 10-17 years in the low-income areas of Pelotas, Brazil. RESULTS: The prevalence of pain in the neck, knee, wrist or hands, and upper back exceeded 15%. Workers in manufacturing had a significantly increased risk for musculoskeletal pain (prevalence ratio [PR]=1.31) and for back pain (PR=1.69), while workers in domestic service had 17% more musculoskeletal pain and 23% more back pain than nonworkers. Awkward posture (PR=1.15) and heavy physical work (PR=1.07) were associated with musculoskeletal pain, while monotonous work (PR=1.34), awkward posture (PR=1.31), and noise (PR=1.25) were associated with back pain. CONCLUSIONS: Musculoskeletal pain is common among working children and teens. Knowledge of occupational risk factors can support actions to restructure work conditions to reduce or eliminate childhood exposure to hazardous conditions. Our results suggest that strategies to prevent musculoskeletal disorders in child workers should be developed. PMID- 16350336 TI - The effects of solvent exposure on memory and motor dexterity in working children. AB - OBJECTIVES: Children working in vehicle spray-painting, mechanical, and other trade workshops are at significant risk of exposure to organic solvents and, as a result, may be at significant risk of developing clinical and subclinical signs of neurotoxicity. This study reports on the association between exposure to solvents and neurobehavioral performance on a number of non-computerized tests for working children exposed to solvents in comparison with nonexposed working children and nonexposed children at school. METHODS: A convenience cross sectional sample of 300 male children aged 10-17 years was recruited for study. The exposed working group and the two nonexposed groups (working and nonworking school) were matched, as far as possible, on geographic location of residence and age. Neurotoxic effects were assessed through a questionnaire and the child's performance on a selection of neurobehavioral tests. RESULTS: Exposed working children scored worse on the overall neurotoxicity symptoms score (mean=6.8; standard deviation [SD]=3.6) compared with the nonexposed working children (mean=1.3; SD=2.0) and school children (mean=1.2; SD=1.8). Analysis of the non computerized neurobehavioral tests demonstrated that exposed working children performed significantly worse than the two nonexposed groups on the motor dexterity and memory tests. Results of the mood test showed that exposed working children were more angry and confused than the nonexposed groups. CONCLUSION: There is an association between exposure to solvents and lower neurobehavioral performance, with significant neurobehavioral deficits among children exposed to solvents in comparison with working children not exposed to solvents and nonworking school children. Memory and motor dexterity appear to be particularly affected in solvent-exposed working children. PMID- 16350338 TI - The Public Health Service in the Panama Canal: a forgotten chapter of U.S. public health. PMID- 16350340 TI - Ethically speaking. PMID- 16350339 TI - Gonzales v. Raich: implications for public health policy. PMID- 16350341 TI - Reality-based planning. PMID- 16350342 TI - Direct reimbursement dental plans. PMID- 16350343 TI - Community and the value of the individual. PMID- 16350344 TI - 10 tips for dealing effectively with employees. PMID- 16350345 TI - Similar mandibular osseous lesions in Tyrannosaurus rex and man. AB - This report identifies several cases of similar-appearing multiple lesions in the mandibles of both humans and the dinosaur Tyrannosaurus rex (T. rex). A diagnosis and potential etiologies are discussed. The appearance of these lesions in prehistoric fossils suggests that this pathology is an ancient affliction which predates humans and our mammalian ancestors. Lytic lesions of the oral structures have occurred in man and higher animals throughout time. The causes range from congenital anomalies, trauma, and infections to benign and metastatic neoplasms. Not only mammals suffer from these conditions; reptiles and birds experience similar diseases. PMID- 16350346 TI - The roadmap to guide patients from diagnosis to acceptance. AB - The journey from diagnosis to treatment acceptance starts with educating patients and building trust. Along the way to the final destination--the dental chair--are three "stops," or critical components to maximizing treatment acceptance. Each stop is equally important and involves different members of the dental team. Just as a car has a steering wheel, engine, and tires, the dental team is comprised of different members with different responsibilities. And the car can only move forward when the team works together. So, are you ready to hit the road to increased production? PMID- 16350347 TI - After-hours emergency coverage by private practitioners in Massachusetts. PMID- 16350349 TI - You are not alone. AB - This article represents the desire of one dentist to share his experiences, strength, and hope with colleagues who have had or are having problems in their lives that can be associated with the abuse of alcohol and habituating or addictive drugs or behaviors. PMID- 16350348 TI - Everything you need to know about the chartless office. AB - One of the biggest buzzwords in the dental industry today is "chartless office." A chartless office vastly improves many of the conventional components in a dental office, including patient management, information flow, and image management. Still, many dentists have raised questions about what is entailed in implementing this system, and the truth is, there are some common misconceptions surrounding this concept of offices without charts. PMID- 16350350 TI - A clinico-pathologic correlation. Glandular odontogenic cyst. PMID- 16350351 TI - Desquamative gingivitis. PMID- 16350352 TI - Inflammatory response. PMID- 16350353 TI - A the challenge of change: may I continue to thrive on it. PMID- 16350354 TI - Increasing compliance with glaucoma therapy: "so, convince me I have something wrong with my eyes". AB - Successful management of primary open-angle glaucoma (POAG) depends heavily on patient compliance with the treatment plan. Studies have clearly shown that early treatment will slow the progress of the POAG. Despite this fact, and even when treatment plans are negotiated with the patient, patient behavior may not be conducive to conserving vision in later life. Poor compliance results from a variety of causes, including lack of understanding of the diagnosis and the progress of POAG. As POAG is a "silent" condition, unless they are aware of visual loss at the point of presentation, POAG patients need to be convinced that there is actually something wrong with them. This paper explores how health care professionals can better understand their patients' response to a diagnosis of POAG. PMID- 16350355 TI - Ocular trauma under the shadow of terror. AB - OBJECTIVES: To determine the prevalence and severity of penetrating eye trauma (PET) in victims of terror attacks in Jerusalem and to determine their specific in-hospital needs; and to evaluate the role of the ophthalmic nurse in helping to treat these patients in wards other than the ophthalmology ward and to determine how the ophthalmic nurse can coordinate care of the patients with other hospital personnel. BACKGROUND: PET is a common cause of significant visual loss. Early treatment is mandatory if maximum vision is to be restored. From January 2000 through February 2003, 468 victims of terror were examined in one of two major hospitals in Jerusalem that treat ocular injuries; 40 (8.5%) of these patients suffered from PET. METHODS: We conducted in our hospital a retrospective chart review of 109 consecutive patients with PET from January 2000 to February 2003; 40 (36.7%) cases were caused by terrorism. Data were collected on demographic characteristics; type, cause and extent of injury; and visual acuity (VA) on admission and at discharge. The terror victims were hospitalized in intensive care, surgical, and ophthalmology wards. RESULTS: Of the patients admitted after terrorist attacks, 64% were male. Arab patients comprised 20% of those admitted. Ninety-five percent of the terror victims with PET required surgery (20% for enucleation, 30% for retinal detachment, and the remaining 45% for other causes). Eighty-two percent of these victims presented with a visual acuity > or = 20/200 in the injured eye (versus 63% in patients admitted with ocular trauma from other causes), and 44% remained blind at discharge (versus 28% from other causes). CONCLUSIONS: Terror victims have more severe ocular injuries than do other patients with PET Ophthalmic nurses must make a special effort to attend to these patients in wards other than the ophthalmology wards and to coordinate their in hospital care and follow-up treatments with hospital personnel in intensive care and surgical wards. PMID- 16350356 TI - Patients with prion disease: planning for eye surgery. AB - Human prion diseases are characterized by rapidly progressive fatal neurodegenerative symptoms. In prion diseases, neural tissues have the greatest potential for infectivity, and ocular tissues are included in the category of high infection risk. The Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO) have published specific infection control guidelines for prion diseases. Standard Precautions are utilized for all patient care. Additional and more stringent measures are indicated when patients with known or suspected prion disease require ocular surgery. Measures include: notification of infection control, operating room, and decontamination personnel; use of disposable instruments whenever possible; special processing for non disposable instruments; identification of tissue and laboratory specimens; and incineration or other appropriate treatment of infectious waste. Clumsiness, vision changes, and myoclonus in association with prion disease may predispose patients to traumatic ocular emergencies. Proactively, procedure areas should utilize a multidisciplinary approach to do the following: identify urgent surgical procedures that are likely to be required by these patients; evaluate cleaning procedures that will result in effective tissue removal; identify critical, non-disposable equipment for which there are no disposable alternatives; identify decontamination methods available to the specific facility as well as to identify which recommended decontamination procedures may not be appropriate for ocular instrumentation; and develop precautionary procedures specifically for decontamination of equipment and instruments. PMID- 16350357 TI - Age-related macular degeneration: answers to some common questions. PMID- 16350358 TI - Color vision testing. PMID- 16350359 TI - The midlatitude North American background aerosol and global aerosol variation. AB - Protocols for the particulate matter (PM) National Ambient Air Quality Standards (NAAQS), and the Regional Haze Rule (RHR) give two complementary definitions for "natural" background airborne particle concentrations in the United States. The definition for the NAAQS derives largely from reported annual averages, whereas the definition for the RHR takes into account the frequency of occurrence of a range of visibility conditions estimated using fine particle composition. These definitions are simple, static representations of background or "unmanageable" aerosol conditions in the United States. An accumulation of data from rural remote sites representing global conditions indicates that the airborne particle concentrations are highly variable. Observational campaigns show weather-related variations, including incidents of regional or intercontinental transport of pollution that influence background aerosol levels over midlatitude North America. Defining a background in North America based on long-term observations relies mainly on the remote-rural IMPROVE network in the United States, with a few additional measurements from Canada. Examination of the frequency of occurrence of mass concentrations and particle components provides insight not only about annual median conditions but also the variability of apparent background conditions. The results of this analysis suggest that a more elaborate approach to defining an unmanageable background could improve the present approach taken for information input into the U.S. regulatory process. An approach interpreting the continental gradients in fine PM (PM2.5) concentrations and composition may be warranted. PMID- 16350360 TI - Natural background visibility and regional haze goals in the Southeastern United States. AB - The goal of the regional haze mitigation program in the United States is to attain "natural conditions" in national parks and wilderness areas by 2064. Results of research investigations on background concentrations of sea salt and biogenic organic matter, of episodic Saharan and Asian dust, and of carbon from natural fires were reviewed to provide a basis for making site-specific estimates of what the concentrations of atmospheric fine particulate matter components might be under natural conditions in the Southeastern United States. Based on this review, rough estimates were made of potential contributions of these aerosol components to natural background visibility. Natural organic particles were the dominant influence on the rate of visibility improvement required to reach natural conditions at an inland, mountainous location, and organic particles and sea salt were the dominant influences on the rate at a coastal location. African dust also had a large episodic effect, but the current regulatory approach is not designed to address episodic background variations. Insufficient data exist to quantify the contributions of wildfires with any detail, although global air pollution modeling provides insight, and their emissions can be locally dominant. Conservative regional refinements to the default natural background estimates do not greatly alter the region-wide rates of reduction of ambient particulate matter concentrations that will be needed to accomplish the first phase of the regional haze program. However, refinements at specific Class I areas may have considerable influence on defining the nature (magnitude and spatial and temporal distribution) of local emission reduction efforts there. PMID- 16350361 TI - A statistical analysis of visibility-impairing particles in federal Class I areas. AB - It is widely agreed that visibility conditions in many Class I areas are impaired to some extent. This paper provides an estimate of the degree of impairment in many of the Class I areas with respect to each of the six haze-forming aerosol classes as described in the supporting documents to the U.S. Environmental Protection Agency Regional Haze Rule. Analyses are performed comparing data from the Interagency Monitoring of Protected Visual Environments network to a modified version of the default natural conditions estimates from the Regional Haze Rule. Uncertainties in the measured annual mean concentrations and biases in the default natural condition estimates because of the effects of geography and meteorology are discussed. It is determined that all of the Class I areas in the contiguous 48 United States are significantly impaired with respect to sulfate aerosols, most of the Class I areas are significantly impaired with respect to nitrate and elemental carbon aerosols, and impairment with respect to organic mass, soil mass, and coarse mass is generally less discernable. No attempt is made to determine adverse impacts with respect to any specific source or group of sources. PMID- 16350362 TI - Spatial, temporal, and interspecies patterns in fine particulate matter in Texas. AB - The Big Bend Regional Aerosol and Visibility Observational (BRAVO) field study was conducted from July to October 1999 and was followed by several years of modeling and data analyses to examine the causes of haze at Big Bend National Park TX (BBNP). During BRAVO, daily speciated fine (diameter <2.5 microm) particulate concentrations were measured at 37 sites throughout Texas. At the primary receptor site, K-Bar Ranch, there were many additional measurements including a "high-sensitivity" version of the 24-hr fine particulate elemental data. The spatial, temporal, and interspecies patterns in these data are examined here to qualitatively investigate source regions and source types influencing the fine particulate concentrations in Texas with an emphasis on sources of sulfates, the largest contributor to fine mass and light extinction. Peak values of particulate sulfur (S) varied spatially and seasonally. Maximum S was in Northeast Texas during the summer, whereas peak S at BBNP was in the fall. Sulfate acidity at BBNP also varied by month. Sources of Se were evident in Northeast Texas and from the Carbon I and II plants. High S episodes at BBNP during BRAVO had several different trace element characteristics. Carbon concentrations at BBNP during BRAVO were probably mostly urban-related, with arrival from the Houston area likely. The Houston artificial tracer released during the second half of BRAVO was highly correlated with some carbon fractions. There was evidence of the influence of African dust at sites throughout Texas during the summer. Patterns in several trace elements were also examined. Vanadium was associated with air masses from Mexico. Lead concentrations in southern Texas have dropped dramatically over the past several years. PMID- 16350363 TI - Directional biases in back trajectories caused by model and input data. AB - Back trajectory analyses are often used for source attribution estimates in visibility and other air quality studies. Several models and gridded meteorological datasets are readily available for generation of trajectories. The Big Bend Regional Aerosol and Visibility Observational (BRAVO) tracer study of July to October 1999 provided an opportunity to evaluate trajectory methods and input data against tracer concentrations, particulate data, and other source attribution techniques. Results showed evidence of systematic biases between the results of different back trajectory model and meteorological input data combinations at Big Bend National Park during the BRAVO. Most of the differences were because of the choice of meteorological data used as input to the trajectory models. Different back trajectories also resulted from the choice of trajectory model, primarily because of the different mechanisms used for vertical placement of the trajectories. No single model or single meteorological data set was found to be superior to the others, although rawinsonde data alone are too sparse in this region to be used as the only input data, and some combinations of model and input data could not be used to reproduce known attributions of tracers and simulated sulfate. PMID- 16350364 TI - Trends in speciated fine particulate matter and visibility across monitoring networks in the Southeastern United States. AB - Trends in fine particulate matter <2.5 microm in diameter (PM2.5) and visibility in the Southeastern United States were evaluated for sites in the Interagency Monitoring of Protected Visual Environments, Speciated Trends Network, and Southeastern Aerosol Research and Characterization Study networks. These analyses are part of the technical assessment by Visibility Improvement-State and Tribal Association of the Southeast (VISTAS), the regional planning organization for the southeastern states, in support of State Implementation Plans for the regional haze rule. At all of the VISTAS IMPROVE sites, ammonium sulfate and organic carbon (OC) are the largest and second largest contributors, respectively, to light extinction on both the 20% haziest and 20% clearest days. Ammonium nitrate, elemental carbon (EC), soils, and coarse particles make comparatively small contributions to PM2.5 mass and light extinction on most days at the Class I areas. At Southern Appalachian sites, the 20% haziest days occur primarily in the late spring to fall, whereas at coastal sites, the 20% haziest days can occur through out the year. Levels of ammonium sulfate in Class I areas are similar to those in nearby urban areas and are generally higher at the interior sites than the coastal sites. Concentrations of OC, ammonium nitrate, and, sometimes, EC, tend to be higher in the urban areas than in nearby Class I areas, although differences in measurement methods complicate comparisons between networks. Results support regional controls of sulfur dioxide for both regional haze and PM2.5 implementation and suggest that controls of local sources of OC, EC, or nitrogen oxides might also be considered for urban areas that are not attaining the annual National Ambient Air Quality Standard for PM2.5. PMID- 16350365 TI - The effects of scrubber installation at the Navajo Generating Station on particulate sulfur and visibility levels in the Grand Canyon. AB - Grand Canyon National Park (GCNP) is a mandatory Class I federal area that is afforded visibility protection under the Federal Clean Air Act. In this paper, we have examined the effects on visibility and particulate sulfur (Sp) at GCNP as a result of reducing sulfur dioxide (SO2) emissions by 90% from the Navajo Generating Station (NGS). Scrubbers were retrofitted to each of the three units at NGS during 1997, 1998, and 1999. The Inter-agency Monitoring of Protected Visual Environments aerosol network database affords us an opportunity to examine trends in Sp and extinction both prescrubber and postscrubber. The NGS impacts GCNP primarily during the winter (December to February). During winter, at times, there are fogs, stratus, and high-relative humidity in the Grand Canyon. When the NGS plume interacts with these fogs and stratus, rapid conversion of SO2 to Sp can occur. A variety of analytical techniques were used, including cumulative frequency plots of Sp and extinction, and chemical mass balance and tracer source apportionment analysis. We also deployed P value statistical analysis of "extreme" Sp values. Before scrubbers were installed, values of Sp approaching 2 microg/m3 were occasionally observed. Because scrubbers have been installed, high levels of Sp have been markedly reduced. Statistical P value analysis suggests that these reductions were significant. Furthermore, we have also observed that Sp has been reduced throughout the cumulative frequency curve during winter by approximately 33% since scrubbers were installed. By contrast, during summer when the NGS impact on the Canyon is minimal, there has been only a relatively small decrease in Sp. PMID- 16350366 TI - Evaluation of multisectional and two-section particulate matter photochemical grid models in the Western United States. AB - Version 4.10s of the comprehensive air-quality model with extensions (CAMx) photochemical grid model has been developed, which includes two options for representing particulate matter (PM) size distribution: (1) a two-section representation that consists of fine (PM2.5) and coarse (PM2.5-10) modes that has no interactions between the sections and assumes all of the secondary PM is fine; and (2) a multisectional representation that divides the PM size distribution into N sections (e.g., N = 10) and simulates the mass transfer between sections because of coagulation, accumulation, evaporation, and other processes. The model was applied to Southern California using the two-section and multisection representation of PM size distribution, and we found that allowing secondary PM to grow into the coarse mode had a substantial effect on PM concentration estimates. CAMx was then applied to the Western United States for the 1996 annual period with a 36-km grid resolution using both the two-section and multisection PM representation. The Community Multiscale Air Quality (CMAQ) and Regional Modeling for Aerosol and Deposition (REMSAD) models were also applied to the 1996 annual period. Similar model performance was exhibited by the four models across the Interagency Monitoring of Protected Visual Environments (IMPROVE) and Clean Air Status and Trends Network monitoring networks. All four of the models exhibited fairly low annual bias for secondary PM sulfate and nitrate but with a winter overestimation and summer underestimation bias. The CAMx multisectional model estimated that coarse mode secondary sulfate and nitrate typically contribute <10% of the total sulfate and nitrate when averaged across the more rural IMPROVE monitoring network. PMID- 16350367 TI - Preliminary evaluation of the Community Multiscale Air Quality model for 2002 over the Southeastern United States. AB - The Visibility Improvement State and Tribal Association of the Southeast (VISTAS) is one of five Regional Planning Organizations that is charged with the management of haze, visibility, and other regional air quality issues in the United States. The VISTAS Phase I work effort modeled three episodes (January 2002, July 1999, and July 2001) to identify the optimal model configuration(s) to be used for the 2002 annual modeling in Phase II. Using model configurations recommended in the Phase I analysis, 2002 annual meteorological (Mesoscale Meterological Model [MM5]), emissions (Sparse Matrix Operator Kernal Emissions [SMOKE]), and air quality (Community Multiscale Air Quality [CMAQ]) simulations were performed on a 36-km grid covering the continental United States and a 12-km grid covering the Eastern United States. Model estimates were then compared against observations. This paper presents the results of the preliminary CMAQ model performance evaluation for the initial 2002 annual base case simulation. Model performance is presented for the Eastern United States using speciated fine particle concentration and wet deposition measurements from several monitoring networks. Initial results indicate fairly good performance for sulfate with fractional bias values generally within +/-20%. Nitrate is overestimated in the winter by approximately +50% and underestimated in the summer by more than -100%. Organic carbon exhibits a large summer underestimation bias of approximately 100% with much improved performance seen in the winter with a bias near zero. Performance for elemental carbon is reasonable with fractional bias values within +/- 40%. Other fine particulate (soil) and coarse particular matter exhibit large (80-150%) overestimation in the winter but improved performance in the summer. The preliminary 2002 CMAQ runs identified several areas of enhancements to improve model performance, including revised temporal allocation factors for ammonia emissions to improve nitrate performance and addressing missing processes in the secondary organic aerosol module to improve OC performance. PMID- 16350368 TI - Reconciliation and interpretation of Big Bend National Park particulate sulfur source apportionment: results from the Big Bend Regional Aerosol and Visibility Observational Study--part I. AB - The Big Bend Regional Aerosol and Visibility Observational (BRAVO) study was an intensive monitoring study from July through October 1999 followed by extensive assessments to determine the causes and sources of haze in Big Bend National Park, located in Southwestern Texas. Particulate sulfate compounds are the largest contributor of haze at Big Bend, and chemical transport models (CTMs) and receptor models were used to apportion the sulfate concentrations at Big Bend to North American source regions and the Carbon power plants, located 225 km southeast of Big Bend in Mexico. Initial source attribution methods had contributions that varied by a factor of > or =2. The evaluation and comparison of methods identified opposing biases between the CTMs and receptor models, indicating that the ensemble of results bounds the true source attribution results. The reconciliation of these differences led to the development of a hybrid receptor model merging the CTM results and air quality data, which allowed a nearly daily source apportionment of the sulfate at Big Bend during the BRAVO study. The best estimates from the reconciliation process resulted in sulfur dioxide (SO2) emissions from U.S. and Mexican sources contributing approximately 55% and 38%, respectively, of sulfate at Big Bend. The distribution among U.S. source regions was Texas, 16%; the Eastern United States, 30%; and the Western United States, 9%. The Carbon facilities contributed 19%, making them the largest single contributing facility. Sources in Mexico contributed to the sulfate at Big Bend on most days, whereas contributions from Texas and Eastern U.S. sources were episodic, with their largest contributions during Big Bend sulfate episodes. On the 20% of the days with the highest sulfate concentrations, U.S. and Mexican sources contributed approximately 71% and 26% of the sulfate, respectively. However, on the 20% of days with the lowest sulfate concentrations, Mexico contributed 48% compared with 40% for the United States. PMID- 16350369 TI - Reconciliation and interpretation of the Big Bend National Park light extinction source apportionment: results from the Big Bend Regional Aerosol and Visibility Observational Study--part II. AB - The recently completed Big Bend Regional Aerosol and Visibility Observational (BRAVO) Study focused on particulate sulfate source attribution for a 4-month period from July through October 1999. A companion paper in this issue by Schichtel et al. describes the methods evaluation and results reconciliation of the BRAVO Study sulfate attribution approaches. This paper summarizes the BRAVO Study extinction budget assessment and interprets the attribution results in the context of annual and multiyear causes of haze by drawing on long-term aerosol monitoring data and regional transport climatology, as well as results from other investigations. Particulate sulfates, organic carbon, and coarse mass are responsible for most of the haze at Big Bend National Park, whereas fine particles composed of light-absorbing carbon, fine soils, and nitrates are relatively minor contributors. Spring and late summer through fall are the two periods of high-haze levels at Big Bend. Particulate sulfate and carbonaceous compounds contribute in a similar magnitude to the spring haze period, whereas sulfates are the primary cause of haze during the late summer and fall period. Atmospheric transport patterns to Big Bend vary throughout the year, resulting in a seasonal cycle of different upwind source regions contributing to its haze levels. Important sources and source regions for haze at Big Bend include biomass smoke from Mexico and Central America in the spring and African dust during the summer. Sources of sulfur dioxide (SO2) emissions in Mexico, Texas, and in the Eastern United States all contribute to Big Bend haze in varying amounts over different times of the year, with a higher contribution from Mexican sources in the spring and early summer, and a higher contribution from U.S. sources during late summer and fall. Some multiple-day haze episodes result from the influence of several source regions, whereas others are primarily because of emissions from a single source region. PMID- 16350370 TI - An innovative methodology for analyzing digital visibility images in an urban environment. AB - A novel methodology combining digital imaging, conventional fixed visibility monitors, and solar radiation monitors has been developed to characterize the visual air quality of the El Paso and Ciudad Juarez urban vista. The authors have found that the digital image quality is reproducible and useful for quantitative analysis of visibility conditions. Regions of interest were selected in images along view paths of interest and values for a contrast variable of interest, typically the coefficient of variation or contrast ratio (CR) for discrete targets, were computed. Both of these indices are bounded at 0 and 1 and are scaled to the "clean day" maxima for a given date, time, and selected view paths. This produces a relative visibility index for various view paths. With the siting of a Belfort (6230A) visibility monitor at a central location, it has been possible to initiate contrast analysis of various targets in current and archived camera images obtained near this monitor. For uniformly "clean" days, as indicated by fine particulate matter observations and visual inspection, the authors have been able to use the extinction coefficient (Bext) derived from the 6230A to put the relative visibility index, based on CR, on an absolute basis in terms of an ideal target located at a given distance. This permits the generation of contrast extinction, Bext/C, for each view path that is independent of the actual target intrinsic contrast (within limits) and allows the comparison of Bext/C along different view paths with other air quality indices. Multiple linear regression was used to derive a relationship between the CR-based Bext/C value and air quality parameters. Visibility attenuation because of sulfate particles was found to have the highest correlation with Bext/C. In addition, solar radiation was observed to be a significant predictor of visibility in the urban region. PMID- 16350371 TI - Scattering cross-section emission factors for visibility and radiative transfer applications: military vehicles traveling on unpaved roads. AB - Emission factors for particulate matter (PM) are generally reported as mass emission factors (PM mass emitted per time or activity) as appropriate for air quality standards based on mass concentration. However, for visibility and radiative transfer applications, scattering, absorption, and extinction coefficients are the parameters of interest, with visibility standards based on extinction coefficients. These coefficients (dimension of inverse distance) equal cross-section concentrations, and, therefore, cross-section emission factors are appropriate. Scattering cross-section emission factors were determined for dust entrainment by nine vehicles, ranging from light passenger vehicles to heavy military vehicles, traveling on an unpaved road. Each vehicle made multiple passes at multiple speeds while scattering and absorption coefficients, wind velocity and dust plume profiles, and additional parameters were measured downwind of the road. Light absorption of the entrained PM was negligible, and the light extinction was primarily caused by scattering. The resulting scattering cross-section emission factors per vehicle kilometer traveled (vkt) range from 12.5 m2/vkt for a slow (16 km/ hr), light (1176 kg) vehicle to 3724 m2/vkt for a fast (64 km/hr), heavy (17,727 kg) vehicle and generally increase with vehicle speed and mass. The increase is approximately linear with speed, yielding emission factors per vkt and speed ranging from 4.2 m2/(vkt km/hr) to 53 m2/(vkt km/hr). These emission factors depend approximately linearly on vehicle mass within the groups of light (vehicle mass < or =3100 kg) and heavy (vehicle mass >8000 kg) vehicles yielding emission factors per vkt, speed, and mass of 0.0056 m2/(vkt km/hr kg) and 0.0024 m2/(vkt km/hr kg), respectively. Comparison of the scattering cross-section and PM mass emission factors yields average mass scattering efficiencies of 1.5 m2/g for the light vehicles and of 0.8 m2/g for the heavy vehicles indicating that the heavy vehicles entrain larger particles than the light vehicles. PMID- 16350372 TI - Improved light extinction reconstruction in interagency monitoring of protected visual environments. AB - The U.S. Environmental Protection Agency (EPA) published the Regional Haze Rule (RHR) in 1999. The RHR default goal is to reduce haze linearly from the baseline period of 2000 through 2004 to natural background in 2064. EPA-recommended method for estimating baseline and natural haze uses the Interagency Monitoring of Protected Visual Environments (IMPROVE) light extinction formula. The IMPROVE formula predicts light extinction from measured aerosol chemical concentrations and estimates of the relative humidity multiplier. On average, the IMPROVE formula overpredicts 6156 nephelometer days (24-hr average measured particle light scattering, bsp) of data by 25%. A new IMPROVED method that reconstructs light extinction using a concentration power law model overpredicts these nephelometer days of data by just 2%. Ignoring the 20% lowest light scattering days, this new IMPROVED formula has a 3% underprediction bias over the 4925 highest nephelometer days with light scattering > or =8 inverse megameters. For comparison, the IMPROVE formula has a 12% overprediction bias for the same days. The IMPROVE formula overprediction averages 77%, 27%, 17%, 9%, and -5% broken down by quintile from lowest to highest nephelometer measured light scattering days. The new IMPROVED formula average overprediction is 21%, -5%, -5%, -2%, and 0%. So, agreement between measured and predicted light scattering improves by modifying the current IMPROVE light extinction formula. PMID- 16350373 TI - Estimating the probability of the public perceiving a decrease in atmospheric haze. AB - Regional haze regulations require progress toward reducing atmospheric haze as measured by particle scattering coefficient of visible light. From a practical perspective, this raises the following question: Given a decrease in extinction, what is the probability that people will notice an improvement in visibility? This paper proposes a quantitative definition of the probability of a perceptible increase in visibility given a decrease in light extinction and a general method to estimate this probability from perception measurements made in the field under realistic conditions. Using data from a recent study of visibility perception by 8 observers, it is estimated that a 2-4 deciview change gives a 67% maximum probability of detecting the improvement. Stated another way, the odds of seeing a difference are at most 2:1 for a change of 2-4 deciviews. A 90% probability requires a change of at least 3.5-7.0 deciviews. The limitations and possible bias in the results of this study are discussed. These results may have a major effect on the cost-benefit analysis of regulatory actions to improve visibility. PMID- 16350374 TI - Methods and results from a new survey of values for eastern regional haze improvements. AB - A contingent valuation survey was designed and implemented in 2003 to elicit the willingness to pay for haze reduction in scenic areas of the Eastern United States. This survey updated and enhanced a portion of a 1988 survey, Chestnut and Rowe's Preservation Values Study, which is commonly used to estimate benefits of haze reduction policies. The 2003 survey incorporated features to address criticisms of the 1988 survey regarding its lack of budget constraints and its simplistic descriptions of changes in visibility. In preparing the new survey, we identified a fundamental error in the 1988 survey regarding the amount of visibility change that respondents were asked to value. When corrected, it reduces the estimates in that study of willingness to pay per unit of visibility change by approximately 40%. More importantly, we estimate that haze reduction benefits based on that study are overstated by approximately 70%. Results from our 2003 survey demonstrate that the distribution of willingness to pay is highly skewed, which implies that a haze reduction policy of which the costs per person are equal to the average willingness to pay would be rejected by between 70% and 80% of the public if subjected to a majority vote. Moreover, estimates of willingness to pay are sensitive to how respondents are reminded of their budget constraints. We find a statistically significant decline in willingness to pay when budgetary reminders are included. Finally, we find evidence that a large portion of the willingness to pay appears to be associated with improvement on the worst condition days. Little additional value appears to be associated with improvements to the majority of days that have haze conditions nearer the average. These survey results have important implications for policy making on haze reduction. PMID- 16350375 TI - Why we do, what we do, when we do it. PMID- 16350376 TI - Basics of employment law: understanding and dealing with adverse employment actions and discrimination in the workplace. AB - In the present setting of fiscal and other constraints placed upon the business world during economically challenging times, there exists both intentional and unintentional opportunities for unfair and illegal treatment of employees. Hospitals and other health care facilities or organizations are not immune. In fact, given the demographics of our field and the general "graying" of our colleagues, a disproportionately higher number of perfusionists reside within the age-protected guidelines established by both federal and state law. Not coincidentally, the pressures on hospitals to stay solvent in the presence of decreasing reimbursement and declining revenues may well create a prescription for unfair treatment of employees. This article will explain the basic concepts of the protections afforded employees by law with regard to freedom from harassment and discrimination in the workplace. Matters related to the hostile workplace environment; age, sex, and race discrimination; and adverse employment actions will be explained and illustrated by applicable case law. In this regard, it is intended that this article will enhance the opportunity for perfusionists to be cognizant of both the express and implied behaviors (both verbal and nonverbal) that may be detrimental to an employment situation, as well as to be cognizant of some of the remedies at law available regarding adverse employment circumstances. PMID- 16350377 TI - Gaseous microemboli and the influence of microporous membrane oxygenators. AB - Gaseous microemboli (GME) are still an unsolved problem of extracorporeal circuits. They are associated with organ injury during cardiopulmonary bypass. Microbubbles of different sizes and number are generated in the blood as the result of different components of the extracorporeal circuit as well as surgical maneuvers. The aim of our study was to observe the behavior of microporous membrane oxygenators to GME in the daily use and in an in vitro model. For the detection of microbubbles, we used a two-channel ultrasonic bubble counter based on 2-MHz Doppler-System with special ultrasound probes. The amount and size of GME were monitored before and after membrane. In 28 scheduled cases with 3 different oxygenators and variability of surgical procedures, we observed the bubble activity in the extracorporeal circuit. In addition, we used an in-vitro model to study the ability of six different oxygenators by removing air in various tests. The oxygenators tested were manufactured with different membrane technologies. The results of our investigations showed varying membrane design lead to a partial removal of GME as well as a change in size and numbers of microbubbles. PMID- 16350378 TI - The effect of temperature and aprotinin during cardiopulmonary bypass on three different methods of activated clotting time measurement. AB - The activated clotting time (ACT) is used frequently for monitoring blood anticoagulant response with heparin before, during, and after cardiopulmonary bypass (CPB). Many cardiac procedures involving CPB require reduction of the patient's blood temperature and use of the serine protease inhibitor, aprotinin. Three different methods of ACT measurement were compared to show the effects of different CPB temperatures and the presence of aprotinin. A total of 42 patients were included in the study: 14 received CPB at 28 degrees C, 14 received CPB at 32 degrees C, and 14 normothermic (37 degrees C) CPB. Within each temperature group, seven received aprotinin. The ACT in each group of patients was measured by a celite activator (C-ACT), a kaolin activator (K-ACT), and a celite, kaolin and glass activator (MAX-ACT). All three methods of ACT measurement showed significant increases (p < .05) in clotting times at hypothermic CPB compared with normothermic groups. During heparinization the C-ACT was significantly increased (p < .05) in the presence of aprotinin. Comparability between the 3 ACT measurement methods showed a very high correlation between C-ACT and K-ACT clotting times (R2 = .8962), and slightly lower correlation between MAX-ACT and C ACT (R2 = .7780), and MAX-ACT and K-ACT (R2 = .7827). All ACT measurements are affected by changes in blood temperature. The C-ACT measurement is prolonged with aprotinin, whereas the MAX-ACT and K-ACT method of measurement in the presence of aprotinin are not significantly altered. It appears that the MAX-ACT produces lower values and may necessitate additional heparin therapy for ACT target values considered safe during CPB. Further study is required from these additional findings. PMID- 16350379 TI - Evidence of systemic cytokine release in patients undergoing cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) causes a systemic inflammatory response syndrome (SIRS), which can progress to an acute lung inflammation known as postperfusion syndrome. We developed a two-phase hypothesis: first, that SIRS, as indicated by elevated cytokines post-CPB, would be correlated with postoperative pulmonary dysfunction (Phase I), and second, that the cytokine interleukin-6 (IL-6) is predominantly released from the heart in CPB patients (Phase II). Blood samples were collected from patients undergoing CPB for elective cardiac surgery. In seven patients (Phase I), arterial samples were drawn before, during (5 minutes and 60 minutes), and after CPB. In 14 patients (Phase II), samples were collected from the coronary sinus, superior vena cava, and a systemic artery at the times indicated previously. Samples were analyzed with enzyme-linked immunosorbent assay: IL-1, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha were assessed in Phase I and IL-6 assessed in Phase II. In Phase I, IL-6, IL-8, and IL-10 were elevated after CPB, but only IL-6 concentrations correlated with lung function. In summary, Phase I data demonstrate that increased IL-6 levels at the end of CPB correlate with reduced lung function postoperatively. In Phase II, IL-6 elevation was similar at all sample sites suggesting that the heart is not the major source of IL-6 production. We suggest that IL-6 be implemented as a prognostic measure in patient care, and that patients with elevated IL-6 after CPB be targeted for more aggressive anti-inflammatory therapy to protect lung function. PMID- 16350380 TI - Variations in battery life of a heart-lung machine using different pump speeds, pressure loads, boot material, centrifugal pump head, multiple pump usage, and battery age. AB - Electrical failure during cardiopulmonary bypass (CPB) has previously been reported to occur in 1 of every 1500 cases. Most heart-lung machine pump consoles are equipped with built-in battery back-up units. Battery run times of these devices are variable and have not been reported. Different conditions of use can extend battery life in the event of electrical failure. This study was designed to examine the run time of a fully charged battery under various conditions of pump speed, pressure loads, pump boot material, multiple pump usage, and battery life. Battery life using a centrifugal pump also was examined. The results of this study show that battery life is affected by pump speed, circuit pressure, boot stiffness, and the number of pumps in service. Centrifugal pumps also show a reduced drain on battery when compared with roller pumps. These elements affect the longevity and performance of the battery. This information could be of value to the individual during power failure as these are variables that can affect the battery life during such a challenging scenario. PMID- 16350381 TI - Transfusion-free cardiopulmonary bypass in Jehovah's Witness patients weighing less than 5 kg. AB - Performing cardiac surgery on pediatric Jehovah's Witness patients is a great challenge for the surgical team and especially for the perfusionist. Jehovah's Witnesses reject blood transfusions on the grounds of their literal interpretation of passages of the Bible. In accordance with this belief, Jehovah's Witnesses feel that it is also forbidden to retransfuse autologous blood that has been separated from their own circulatory system. We report the use of cardiopulmonary bypass (CPB) during open-heart surgery in three infants with a body weight of 4.5 kg, 3.5 kg, and 3.1 kg, respectively, without transfusion of blood components. A small-volume CPB circuit with a priming volume of 200 mL, including the arterial line filter, was designed to decrease the degree of hemodilution. A dedicated pediatric heart lung machine console with remote pump heads and intensive blood conservation efforts allowed the operation without the use of donor blood. The CPB circuits were primed with crystalloid solution only. The procedures were performed in normothermia or in moderate hypothermia. Pre-CPB hemoglobin levels were 10.8 g/dL, 10.6 g/dL, and 8.5 g/dL. The hemoglobin concentrations measured during CPB ranged from 5.9 to 6.5 g/dL, 6.4 to 6.8 g/dL, and 5.5 to 5.9 g/dL, respectively. The patients did not receive any blood or blood products during their entire hospital stay. PMID- 16350382 TI - A prospective comparison of the platelet sequestration ability of three autotransfusion devices. AB - Although current autotransfusion devices have platelet sequestration capabilities, each has a unique technology to achieve the final platelet product. The purpose of this study was to assess the quality and quantity of platelets sequestered by three different autotransfusion devices. The three commercially available autotransfusion devices evaluated were Fresenius C.A.T.S (closed spiral chamber), Cobe BRAT 2 (Baylor bowl), and Haemonetic Cell Saver 5 (Latham bowl). Platelet sequestration was preformed in the automatic mode following the manufacturer's recommended sequestration protocols. The total number of platelets sequestered, percent recovery, and percent platelet function were assessed. Each device behaved similarly. There was a 2- to 3-fold increase in platelet count compared with baseline with only approximately 50-60% recovery, whereas there was approximately a 10% decrease in platelet function after processing compared with baseline. No statistical difference was noted in platelet function between the respective machines. However, there was a significant loss of platelet function observed with the actual process regardless of autotransfusion device used. PMID- 16350383 TI - Clinical performance and biocompatibility of novel hyaluronan-based heparin bonded extracorporeal circuits. AB - We tested documented in vitro and ex vivo advantages of novel hyaluronan based heparin bonded extracorporeal circuits in a prospective randomized study. During the period from June until September 2005, 40 patients undergoing reoperation for coronary artery bypass grafting were allocated into two equal groups (n = 20): Group 1 was treated with hyaluronan-based heparin-bonded circuits and group 2 was treated with uncoated control circuits. Complete blood count, fibrinogen, albumin, C3a, interleukin-2 levels, and thromboelastographic data were documented after induction of anesthesia (T1) and heparin administration before cardiopulmonary bypass (CPB) (T2), 15 minutes after initiation of CPB (T3), before cessation of CPB (T4), 15 minutes after reversal with protamine (T5), and the first postoperative day at 8:00 a.m. (T6). Hollow fibers were collected for consecutive biomaterial analysis by optical and scanning electron microscopy (SEM). Desorbed protein deposition on fibers was compared by spectrophotometry. Leukocyte counts were lower in T4-T6 in group 1 (p < .05). Platelet counts demonstrated significant differences at T4 and T5 in coated group (p < .05). Albumin and fibrinogen levels were better preserved in Group 1 at T4, T5 and T4, T6, consecutively (p < .05). C3a and IL-2 levels were lower at T3-T5 and T4-T5 in intervention group (p < .05). Postoperative hemorrhage was 412 +/- 50 mL in group 1 and 684 +/- 50 ml in group 2 (p < .05). Respiratory support time was shorter in group 1 versus control (p < .05). Platelet adhesion was significantly lower in intervention group. Amount of desorbed protein was 1.44 +/- 0.01 mg/dL in group 1 and 1.94 +/- 0.01 mg/dL in control (p < .05). SEM and spectrophotometry demonstrated better surface preservation in the hyaluronan coated group. Novel hyaluronan-based heparin-bonded circuits reduce platelet adhesion-aggregation and protein adsorption and provide better perioperative clinical parameters through platelet, albumin, and fibrinogen-sparing effects. PMID- 16350384 TI - Use of bivalirudin as an anticoagulant during cardiopulmonary bypass. AB - Bivalirudin is a short-acting direct thrombin inhibitor that has been used in cardiac surgical patients with heparin-induced thrombocytopenia (HIT) or suspected HIT. Although no direct thrombin inhibitor is indicated for anticoagulation during cardiac surgery in patients with heparin-induced thrombocytopenia (HIT) or suspected HIT, use of heparin-alternatives are increasing as the awareness of HIT increases. Reports of anticoagulation with bivalirudin are sporadic, however, with variable dosing and management strategies. In this report, we describe our management techniques for cardiopulmonary bypass with bivalirudin based upon our personal experience. Although the reported clinical experience with bivalirudin in cardiac surgery is reviewed, operative techniques for the perfusionist/surgeon team are discussed in detail. We recognize that the use of bivalirudin during cardiopulmonary bypass is evolving and modifications of technique will undoubtedly occur as further data and experience accumulate. PMID- 16350385 TI - Cardiopulmonary bypass using argatroban as an anticoagulant for a 6.0-kg pediatric patient. AB - A patient was born with transposition of the great arteries, double-outlet right ventricle, interrupted aortic arch, and a ventricular septal defect and underwent a Damus-Kaye-Stansel procedure with a modified Blalock-Taussig shunt at 14 days old. Three months later, this patient presented with hypoxia and bradycardia was found to have a thrombus present in the main pulmonary artery extending to right pulmonary artery. After initiation of thrombolytic therapy, the patient became severely hypoxic and required the institution of extracorporeal membrane oxygenation. As the result of unknown heparin resistance independent of adequate antithrombin III levels, argatroban therapy was used to achieve desired anticoagulation. The patient was taken to the operating room and converted to conventional cardiopulmonary bypass once adequate activated clotting times were achieved using argatroban. This case report summarizes the use of argatroban as an anticoagulant for a 6.0-kg pediatric patient undergoing cardiopulmonary bypass. PMID- 16350386 TI - The coronary baroreflex in humans. AB - Previous studies have identified the presence of coronary baroreceptors in animal models. We set up a study to explore the presence of coronary baroreceptors in humans, which was performed with isolated, graded aortic root perfusion in patients during cardiopulmonary bypass. With ethical approval 12 patients with normal coronary arteries, aged 58-75 (mean 69) years undergoing mitral valve surgery were recruited to the study with informed consent. Those with aortic valve incompetence, coronary, or peripheral artery disease and diabetes mellitus were excluded. They were randomized to have their coronary perfusion pressure set low at 50 mmHg for 90 seconds and then adjusted high to 80 mmHg for 90 seconds (group L-H) or the reverse sequence (group H-L). Average arterial pressure and approximately constant systemic flow over 30-second periods were used to calculate vascular resistance (SVR). The first six experiments followed initiation of cardiopulmonary bypass and aortic clamping but before the delivery of cold blood cardioplegia; the blood temperature for these experiments was kept at 32 degrees C. The remaining six were conducted prior to removal of the aortic cross clamp at 37 degrees C. Coronary sinus blood samples were analyzed to exclude myocardial ischemia. Coronary sinus blood samples showed insignificant variation in oxygen saturation, lactate, and troponin T. Three patients were excluded because of unstable blood pressure. In the (L-H) group SVR reduced in 4 of 4 remaining patients (mean -9.4%, range -3.9 to -19.6%). In the (H-L) group SVR increased in three patients (mean +2.0%, range 1.1 to 3.7%) but decreased in two (-8.9% and -15.8%). These preliminary results, although not statistically different, suggest the presence of coronary baroreceptors in humans. The reflex vascular responses are similar to those previously reported in animal models. PMID- 16350387 TI - The use of hydroxy-ethyl starch sedimentation for autologous buffy coat preparation. AB - The use of hydroxy-ethyl starch (HES) has been used in the cord blood banking industry for the separation of blood into its individual components. The focus of this study is to examine whether the use of HES is feasible in obtaining a buffy coat for the use in the construction of platelet gels. Blood was collected from seven canines into anticoagulant citrate dextrose at a ratio of 1:8. A sample of whole blood was used to obtain initial cell counts and a base line Thrombelastograph (TEG). 6% Hespan was added to the whole blood and allowed to sediment for 60 minutes. Cell counts and TEGs were performed on the resulting layers: red blood cells (RBCs), buffy coat (BC), and plasma. The results of this study (see Table 1) suggest that the gravity sedimentation of whole blood with HES can significantly reduce the RBCs in the BC and plasma layers while increasing the platelet count in the plasma layer. These data also suggest that HES does not affect the clotting capabilities of the BC and plasma layers. PMID- 16350388 TI - A description of a prototype miniature extracorporeal membrane oxygenation circuit using current technologies in a sheep model. AB - In the United States, standardization of neonatal extracorporeal membrane oxygenation (ECMO) circuit was achieved during the 1980s. Since that time, the consoles and components of the ECMO circuit have remained fundamentally unchanged (bladder, rollerpump, silicone membrane oxygenator). Extracorporeal technology, however, has witnessed many significant advancements in components during the past two decades. These new technologies have characteristics that may improve outcomes when applied in the ECMO arena. Understanding how these technologies perform in long-term applications is necessary. Therefore, the purpose of this project is to evaluate the performance of a miniature ECMO circuit consisting of current generation technologies in an animal model. An ECMO circuit (prime volume 145 mL) was designed that included a hollow fiber oxygenator and a remote mounted centrifugal pump. All circuit tubing and components were surface coated. Three sheep (approx 13 kg) were placed on ECMO using standard neck cannulation techniques and maintained according to clinical protocols. Technical implementation, oxygenator function, and hematological parameters were accessed. Duration of ECMO was 20, 48, and 58 hours. There was no evidence of oxygenator failure, as measured by pressure drop and oxygen transfer, in any of the procedures. No plasma leak was observed in any oxygenators. Platelet count trended downward after 24 hours. Visual inspection after ECMO showed very little evidence of gross thrombosis. This ECMO circuit design departs dramatically from the typical North American systems. The use of this console and components facilitated a 70% reduction in priming volume over a traditional ECMO circuit. Further investigations should be conducted to determine if circuit miniaturization can reduce the morbidity associated with blood product consumption and the bloods contact with the artificial surfaces of the ECMO circuitry. PMID- 16350390 TI - Continuity of cardiovascular care. PMID- 16350389 TI - The emergency use of recombinant hirudin in cardiopulmonary bypass. 1999. PMID- 16350391 TI - Hypothyroidism and cardiovascular disease. PMID- 16350392 TI - Identification of new papillomavirus types. AB - The identification of papillomavirus DNA sequences in tissue samples using polymerase chain reaction (PCR) amplification, has led to the association of these infections to a multiplicity of clinical manifestations. The cloning and sequencing of PCR-amplified products has, to date, resulted in the identification of more than 300 putative "new" papillomavirus types. The methods used to identify these unknown papillomavirus sequences are described here. The CP, FAP, and GP primers are used for PCR amplification, followed by cloning and sequencing of the amplicons. Sequence comparisons and the interpretation of DNA sequence identities are discussed. Details of defining a new papillomavirus type and of the recently approved taxonomic classification system for the Papillomaviridae are given. PMID- 16350393 TI - Identification of HPV variants. AB - The vast majority of anogenital carcinomas are caused by high-risk human papillomaviruses (HPVs), and among Western nations HPV-16 is usually the most predominant cancer-associated type. As a DNA virus, HPV type 16 has a relatively stable genome that is believed to have co-evolved with its host over the millennia. Nevertheless, among the "wild" populations of HPV-16 that are circulating, a large number of variants have been identified, and these may have considerably different pathogenic potentials. In this chapter, methods for screening and characterizing HPV-16 sequence variants are described. In particular, we describe methods for the identification of variation within the HPV-16 E5 open reading frame and for the detection of the nt 131 A-->G mutation of the E6 ORF, using restriction fragment length polymorphism assays. In addition, we describe approaches for DNA sequencing and analysis. Such methods are likely to be of particular interest to those involved in epidemiological investigations of virus transmission and pathogenicity studies. PMID- 16350394 TI - Histochemical analysis of cutaneous HPV-associated lesions. AB - Hematoxylin and eosin (H&E) staining of cutaneous warts is presented to illustrate the practical methods utilized for histochemical analysis of cutaneous human papillomavirus-associated lesions. Every step of the staining procedure, from sampling of the specimens to microscopic examination of the stained sections, is detailed with reference to the recent achievements in this field. PMID- 16350395 TI - Histological analysis of cervical intraepithelial neoplasia. AB - A wide interobserver variation is seen even among competent histopathologists in the routine diagnosis of cervical intraepithelial neoplasia (CIN). As a result, early detection of low-grade CIN (CIN 1) lesions, in particular, remains a major challenge both in routine diagnosis and in cervical screening. In this chapter, the salient diagnostic features of human papillomavirus infection and CIN lesions are demonstrated. PMID- 16350396 TI - Detection of papillomavirus proteins and DNA in paraffin-embedded tissue sections. AB - The key events during the papillomavirus life cycle can be mapped in infected tissue samples by antibody detection and in situ hybridization. The ease of immuno-detection varies for different proteins and is dependent on antigen availability. Epitope exposure is sometimes necessary, because the antigen may become masked after formalin fixation and paraffin embedding of the infected tissue. Visualization of both nucleic acid and protein targets can be done simultaneously by combining in situ hybridization and immuno-detection methods. PMID- 16350397 TI - Detection and quantitation of HPV gene expression using real-time PCR. AB - Human papillomaviruses (HPVs) are known to be etiological agents of cervical cancer and have been found in 99.7% of women with high-grade (HG) cervical intraepithelial neoplasia (CIN) precancer. Testing of high-risk HPV (HR-HPV) has been proposed as a way of improving cervical screening, especially for women with low-grade (LG) Papanicolaou (Pap) smears. In this chapter, real-time quantitative polymerase chain reaction (PCR) methods that can be used to investigate the expression of HPV 16 early genes in HG or LG precancer are demonstrated. Detecting the expression of early HPV genes in conjunction with the Pap smear may improve the specificity of identifying LG precancers that are associated with high risk of progression. PMID- 16350398 TI - Analysis of p16INK4a and integrated HPV genomes as progression markers. AB - Most cervical cancers are preventable when the precursor lesions are detected in time. Human papilloma viruses (HPVs) are the main risk factors for cervical cancer development, but there is a high percentage of healthy women infected with HPV that never develop a lesion. Only a small percentage of low-grade dysplasias finally grow out to invasive cancer. Several biomarkers can be used to identify lesions at risk for malignant progression. Overexpression of p16INK4a is induced by the viral oncoprotein E7 and distinguishes dysplastic lesions from benign changes. Integration of human papillomavirus DNA into the host genome is mainly found in high-grade dysplastic lesions and invasive cancers, and points to an increased progression potential. PMID- 16350399 TI - Use of biomarkers in the evaluation of CIN grade and progression of early CIN. AB - The treatment of cervical intraepithelial neoplasia (CIN) depends on the evaluation of CIN grade. The grading of CIN is however problematic, as intra- and interobserver reproducibility of CIN-grade evaluation among pathologists is not perfect. There are also difficulties in reliably distinguishing CIN from nonneoplastic lesions, and over- or undertreatment can be the result. These points suggest a need for adjuvant methods that can distinguish CIN from nonneoplastic lesions, and can distinguish different CIN grades and predict the risk of progression of early CIN1 and -2 lesions. This chapter describes the use of biomarker-related methods for the diagnosis and prognostic evaluation of patients with CIN1 and CIN2. As CIN involves the progressive dysfunction of proliferation and differentiation activities in cervical epithelial cells, we have concentrated in this chapter on demonstrating the utility of proliferation- and differentiation-related biomarkers. PMID- 16350400 TI - HPV DNA detection and typing in cervical scrapes. AB - Polymerase chain reaction (PCR)-based assays that use consensus primers to detect DNA of a broad spectrum of human papillomavirus (HPV) types in a single assay belong to the most frequently used methods to detect HPV in clinical specimens. Here, we describe in detail one of these assays, the so-called GP5+/6+ PCR method, which can be used to detect and type HPV DNA in crude extracts of cervical scrapes and biopsy specimens. Following PCR with GP5+ and GP6+ primers, the latter of which is biotinylated at its 5' end, the presence of DNA of any of the high-risk genotypes can easily be determined by an enzyme immunoassay (EIA). In this assay, PCR products are captured in streptavidin-coated wells of a microtiter plate, denatured by alkaline treatment, and hybridized to cocktails of digoxigenin-labeled oligonucleotides specific for high-risk or low-risk HPV types. The resulting hybrids can then be detected by alkaline phosphatase conjugated anti-digoxigenin polyclonal antibodies and substrate followed by optical density reading. Subsequently, EIA-positive PCR products can be typed by a reverse line blot genotyping procedure that, using a miniblotter device, enables typing of up to 39 samples with specific oligonucleotide probes for 37 different HPV (sub)types in a single assay. PMID- 16350401 TI - HPV DNA detection and typing in inapparent cutaneous infections and premalignant lesions. AB - Epidemiological studies, which address the role of human papillomavirus (HPV) in the pathogenesis of (pre)malignant cutaneous lesions, focus on the HPV B1 subgroup comprising the so-called epidermodysplasia verruciformis (EV)-associated HPV types. To detect and type HPV DNA in human materials, Polymerase Chain Reaction (PCR)-based assays are used. In this chapter, a nested, broad-spectrum PCR method using a mixture of primers and a type-specific PCR using specific primers are described. The broad-spectrum PCR detects the B1 subgroup of HPV types. HPV typing is performed by sequence analysis of the PCR product. The type specific PCR detects and types HPV 5a, 8, 15, 17, 20, 24, 36, and 38. These HPV types are representative of the B1 subgroup, because they are evenly distributed over the phylogenetic tree of the B1 subgroup. PMID- 16350402 TI - Establishing HPV-containing keratinocyte cell lines from tissue biopsies. AB - The generation of cell strains and established cell lines from human papillomavirus (HPV)-infected cervical biopsies and ano-genital warts is best achieved by the application of conventional protocols for keratinocyte cell culture. The optimal protocol that permits growth at clonal density and serial cultivation involves the use of an inactivated 3T3 fibroblast feeder layer and medium supplemented with hydrocortisone and fetal bovine serum. Modifications in terms of additives and serum concentration are required for optimal culture of neoplastic and malignant HPV-containing keratinocytes. PMID- 16350403 TI - Differentiation of HPV-containing cells using organotypic "raft" culture or methylcellulose. AB - The study of human papillomaviruses (HPVs) has been challenging due to the differentiation-dependent aspects of their productive life cycles. The use of HPV virions, isolated from tissues, to study viral pathogenesis has been complicated due to the low numbers of HPV virions synthesized and inefficient infection of cells in tissue culture. As an alternative approach, genetic methods have been developed to study the papillomavirus life cycle in its natural host, human keratinocytes. Techniques have been developed to transfect keratinocytes with cloned HPV DNA and to isolate cell lines that maintain viral DNA as extra chromosomal elements. Since the productive phase of the HPV life cycle is dependent on differentiation, in vitro tissue-culture models have also been used to recapitulate epithelial differentiation. Differentiation in organotypic raft cultures as well as upon suspension in semi-solid media have been used to study both early and late stages of the viral life cycle. PMID- 16350404 TI - Propagation of infectious, high-risk HPV in organotypic "raft" culture. AB - The organotypic (raft) culture system has been used to develop an in vitro system that is capable of reproducing the entire human papillomavirus (HPV) life cycle, including virion morphogenesis. This system utilizes HPV-containing cell lines that are either derived from biopsies or created by the transfection of keratinocytes with HPV genomic DNA. When grown as raft cultures, these lines allow for a detailed study of all stages of the viral life cycle. In this chapter, we describe in detail how to (1) culture keratinocytes, (2) electroporate primary keratinocytes with HPV DNA, (3) detect episomal HPV genomes by Southern (DNA) blotting, (4) grow organotypic raft cultures, (5) isolate HPV, and (6) perform in vitro infectivity testing. PMID- 16350405 TI - The cottontail rabbit papillomavirus model of high-risk HPV-induced disease. AB - Animal models are essential to study the pathogenesis of papillomavirus infection and develop strategies for treatment and prevention. This review details use of the cottontail rabbit papillomavirus (CRPV)-laboratory rabbit model. The protocols describe how to infect rabbits with CRPV DNA or CRPV virus to induce papillomas. They also describe the design and analysis of genetic pathogenesis experiments, prophylactic and therapeutic intervention experiments, and malignant progression experiments. PMID- 16350406 TI - Studying the HPV life cycle in 3A trophoblasts and resulting pathophysiology. AB - Human papillomavirus (HPV) productive infection has been long considered to be restricted to the squamous epithelium. However, we have demonstrated that both HPV type 16 (HPV-16) and type 31b (HPV-31b) productively replicate in a trophoblast cell line, 3A. Trophoblasts are an important cell type, as these are the cells of the placenta that directly contact maternal tissues. There are a number of studies that suggest that HPV infection may be found commonly in placental material and might be linked with spontaneous abortions. This chapter describes the preparation and transfection of HPV genomes into 3A cells, suggests ways to analyze viral replication and transcription, and describes how to assay the effect of HPV infection on the ability of the trophoblasts to bind endometrial cells. PMID- 16350407 TI - Analysis of HPV transcription by RPA. AB - Human papillomavirus (HPV) transcription is a complex process using multiple promoters, splices, and polyadenylation sites to create polycistronic transcripts capable of encoding the known and putative viral genes at the correct time and place throughout the differentiation-dependent life cycle. The ribonuclease protection assay (RPA) provides a flexible and convenient tool to study the behavior of HPV transcripts under a variety of cellular conditions and treatments, or in response to genetic mutations. Using a known cloned DNA as a template, an antisense RNA probe is generated and hybridized to the sample RNA. After digestion with ribonucleases (RNases), the fragments of the probe protected by the sample are examined by gel electrophoresis. With the proper design of the probe template, information about promoter usage, splicing, transcript levels, and other parameters can be accurately, simply, and quantitatively measured throughout the HPV life cycle. PMID- 16350408 TI - Analysis of regulatory motifs within HPV transcripts. AB - Papillomavirus late gene expression is highly dependent on host epithelial cell differentiation, such that capsid proteins are produced only in differentiating cells. Several papillomaviruses contain negative regulatory elements (NREs), that is, short regions of late transcripts that interact with host cellular RNA processing factors to prevent capsid protein synthesis in undifferentiated cells. In this chapter, the human papillomavirus (HPV)-16 NRE will be used as an example to show how cis-acting RNA regulatory elements can be identified and mapped using transient transfection of reporter gene constructs. The use of reporter gene assays is also readily applicable to the identification and characterization of novel promoters and other regulatory sequences in HPV DNA. In vitro RNA-protein binding techniques, including ultraviolet crosslinking, electrophoretic mobility shift assay, and affinity purification of RNA binding proteins, will also be described, again using the HPV-16 NRE as an example. These techniques may be used to identify cellular proteins that bind the NRE, allowing its mode of action to be deduced. They may also be used to study interactions between host cellular proteins and other protein-binding motifs on HPV mRNA. These interactions are important for the regulation of HPV gene expression, and have key roles in splicing, polyadenylation, mRNA export, stability, and translation. PMID- 16350409 TI - Detection of HPV transcripts by nested RT-PCR. AB - Human papillomaviruses (HPVs) are etiologic for the development of cervical cancer and its precursor lesions, cervical intraepithelial neoplasia (CIN). Nearly all cervical carcinomas (CaCx) harbor HPV DNA, but the presence of HPV alone is not indicative of the future development of neoplasia. While both normal and abnormal smears may harbor HPV DNA, the detection of HPV mRNA is associated, although not exclusively, with abnormal cytology. Recent observations suggest women with a high HPV viral load are at a significantly greater risk for CIN development--particularly those infected with high-risk (HR) HPV types, such as HPV type 16 (HPV 16). Thus, assays capable of detecting HPV transcripts may have useful prognostic value and could be utilized to identify biological markers for progression to high-grade cervical disease. This chapter describes the nested reverse transcriptase (nRT)-polymerase chain reaction (PCR) methods developed in our laboratory for the detection of the majority of all early region HPV-16 transcripts. PMID- 16350410 TI - Detection and quantitation of HPV DNA replication by Southern blotting and real time PCR. AB - This provides a brief introduction into the mechanism of DNA replication by the E1 and E2 proteins and describes the traditional Southern blotting technique that is used to monitor E1- and E2-mediated DNA replication. It also includes a novel real-time polymerase chain reaction (PCR) approach for monitoring E1- and E2 mediated DNA replication that has enhanced sensitivity and quantitation compared with Southern blotting, and a discussion of when to use the Southern blotting and real-time PCR techniques. PMID- 16350411 TI - Analysis of E7/Rb associations. AB - The product of the early gene E7 is one of the major transforming proteins of human papillomaviruses (HPVs). It exerts its activity by associating with and altering the biological functions of several cellular proteins involved in the control of fundamental events, such as cell proliferation and apoptosis. The best characterized activity of E7 from HPV type 16, the most frequently detected type in cervical cancer, is its ability to bind and induce degradation of the tumor suppressor retinoblastoma protein (pRb) via the ubiquitin pathway. pRb plays a key role in cell-cycle control by negatively regulating, via direct association, the activity of several transcription factors, including members of the E2F family. The neutralization of pRb functions mediated by E7 results in constitutive activation of the transcription factors, with consequent loss of cell-cycle control. Several studies have shown that the oncogenic potential of a specific HPV type is dependent on the efficiency of E7 in targeting pRb. In this chapter, we describe two methods to measure the efficiency of the E7 proteins from different HPV types in neutralizing the pRb functions. The first one, the plate-binding assay, allows the determination of the pRb binding affinity of E7 proteins, while the second one permits the analysis of their impact on the pRb pathway in intact cells. PMID- 16350412 TI - Transformation assays for HPV oncoproteins. AB - A cornerstone of human papillomavirus (HPV) research was the demonstration that those HPV types associated with the development of cervical cancer encode two potent oncoproteins, while those HPV types associated with only benign lesions do not. Thus both HPV-16 E6 and E7 will transform established rodent cells and will efficiently cooperate with other activated oncogenes in the transformation of primary rodent cells. In addition, the virus also encodes for the E5 oncoprotein, which also possesses a weaker transforming activity in established rodent cells. In this chapter we describe how the transforming activities of the HPV oncoproteins can be assessed. PMID- 16350413 TI - Analysis of adeno-associated virus and HPV interaction. AB - It is slowly becoming accepted that adeno-associated virus (AAV) is another significant factor involved in cervical carcinogenesis. However, unlike human papillomavirus (HPV), which is positively associated with cervical cancer, AAV is negatively associated with this cancer. This negative association appears to be through a direct and complex bi-directional interaction between AAV and HPV. Essentially all assays used for studying HPV can be used for studying the AAV-HPV interaction. This is because both viruses are productive in the same tissue, the stratified squamous epithelium (skin). Their relationship can be studied on the level of the complete virus and their complete life cycle using the organotypic epithelial raft culture system, which generates a stratified squamous epithelium. Their relationship can be studied in various other tissue-culture models measuring oncogenic potential. Their interaction can also be studied on the component level, as both protein-protein and protein-DNA interactions are known. Their relationship has even been studied using transgenic animals. The AAV-HPV relationship can be broken down into two halves--AAV-encoded products, which affect HPV biology, and HPV-encoded products, which affect AAV biology. To date, the former are much better studied than the latter. The rep gene and its largest product, Rep78, are responsible for most of AAV's effects upon HPV. This chapter largely focuses on AAV's effect on the HPV life cycle. PMID- 16350414 TI - In vitro assays of substrate degradation induced by high-risk HPV E6 oncoproteins. AB - The high-risk mucosal human papillomavirus E6 proteins were the first viral proteins that were shown to use the ubiquitin proteasome pathway for the inactivation of their cellular target proteins. The first substrate to be identified was the p53 tumor suppressor protein, and since then many other substrates for E6-induced degradation have been described. All of these require the presence of high-risk mucosal E6 together with the E1, E2, and E3 enzymes of the ubiquitin pathway. This activity of E6, although complex, is nonetheless amenable to in vitro analysis. Many different protocols have been described over the years for performing these assays. In this chapter we describe the most easily used and robust procedure that is routinely used in our laboratory. PMID- 16350415 TI - Measuring the induction or inhibition of apoptosis by HPV proteins. AB - Many viral proteins influence the cellular pathways that control cell proliferation and cell death. Some viral proteins trigger apoptotic cell death, and this may be important in host defense and viral spread. In other cases, viral proteins inhibit apoptosis. In this chapter, we will describe some of the methods that can be used to investigate the induction or inhibition of apoptosis by papillomavirus proteins. PMID- 16350416 TI - Codon optimization of papillomavirus genes. AB - Early and late genes of human and animal papillomaviruses show a codon composition seemingly unfavorable for expression in mammalian cells. It remains unclear how the viruses manage to achieve high levels of late gene expression during the viral life cycle. One possible solution could be that the availability of certain t-RNAs changes with progressing stages of cellular differentiation. Previous studies have demonstrated that modification of codon usage of papillomavirus late (L1 and L2) and early genes (E7) can overcome poor expression of these proteins both in transient and in stable expression systems. This was shown not only for human but also for plant cells. Two strategies can be employed to alter codon usage: elimination of only those codons that are rarely used in a particular expression system, or exchange of all possible codons by the ones most frequently used. Currently, there are two protocols for codon modification--a template-less polymerase chain reaction (PCR)-based protocol, in which very long overlapping oligodeoxynucleotides are used in an overlap-extension reaction, or a ligase chain reaction, in which shorter oligodeoxynucleotides are fused together after an annealing procedure. Both methods are presented and discussed. PMID- 16350417 TI - Generation of HPV pseudovirions using transfection and their use in neutralization assays. AB - It has recently become possible to generate high-titer papillomavirus-based gene transfer vectors. The vectors, also known as papillomavirus pseudoviruses (PsV), have been useful for studying papillomavirus assembly, entry, and neutralization, and may have future utility as laboratory gene-transfer tools or vaccine vehicles. This chapter outlines a simple method for production of PsV and their use in a high-throughput papillomavirus neutralization assay. The production method is based on transfection of a 293 cell line, 293TT, engineered to express high levels of SV40 large T antigen. The cells are co-transfected with codon modified papillomavirus capsid genes, L1 and L2, together with a pseudogenome plasmid containing the SV40 origin of replication. Pseudogenome encapsidation within L1/L2 capsids is largely sequence independent, and plasmids entirely lacking PV sequences can be packaged efficiently, provided they are less than 8 kilobases in size. Non-infectious virus-like particles (VLPs) can also be produced after transfection of 293TT cells with L1 alone. Efficient purification of the PsV or VLPs is achieved by Optiprep (iodixanol) density gradient ultracentrifugation. Using these methods, it is possible to produce highly purified PsV with yields of at least 10(9) transducing units from a single 75-cm2 flask of cells. PsV encapsidating a secreted alkaline phosphatase (SEAP) reporter plasmid were used to develop a high-throughput in vitro neutralization assay in a 96-well plate format. Infection of 293TT cells is monitored by SEAP activity in the culture supernatant, using a highly sensitive chemiluminescent reporter system. Antibody-mediated PsV neutralization is detected by a reduction in SEAP activity. The neutralization assay has similar analytic sensitivity to, and higher specificity than, a standard VLP-based enzyme-linked immunosorbent assay (ELISA). PMID- 16350418 TI - Generation and applications of HPV pseudovirions using vaccinia virus. AB - This chapter outlines the generation and application of human papillomavirus type 33 (HPV 33) pseudovirions. These pseudovirions are structurally indistinguishable from native virions and are therefore valuable tools for the study of papillomavirus/cell interactions. The method describes (1) the construction of vaccinia viruses recombinant for the major and minor HPV capsid proteins, L1 and L2, respectively, (2) the transfection of Cos7 cells with a marker plasmid replicating to high copy numbers, (3) the expression of L1 and L2 using the vaccinia virus expression system, (4) the extraction, purification, and analysis of HPV-33 pseudovirions, (5) pseudoinfection assays, (6) pre- and post-attachment neutralization of pseudovirions, and (7) the use of inhibitors for study of binding and uptake of pseudovirions. The methods described have been successfully adopted for HPV 16 and 18 and may thus be applied for other HPV types, too. PMID- 16350419 TI - Fibrous connective tissue between Muller's muscle and the palpebral conjunctiva as a reinforcement structure and a natural barrier for the upper eyelid. AB - This study was performed to illustrate and discuss the significance of fibrous connective tissue between the Muller's muscle and the palpebral conjunctiva. Nine upper eyelids of 6 Oriental cadavers were microscopically examined; ages at death ranged from 72 to 91 years. Tissue of the posterior lamella of the upper eyelid was removed without the orbital septum and orbital fat. Removed eyelids were incised perpendicularly at the center of the eyelid. After pretreatment, sliced sections were stained with Hematoxylin and Eosin and examined microscopically. A thick fibrous connective tissue was found to exist between the Muller's muscle and the palpebral conjunctiva. The connective tissue continued proximally to the intermuscular transverse ligament and was distally attached to the posterior site of the upper aspect of the tarsus. All cases showed infiltration of lymphocytes from the conjunctiva; however, these were completely blocked by the fibrous connective tissue and never reached Muller's muscle. This connective tissue supports eyelid traction and is a natural barrier for the Muller's muscle against conjunctivitis. PMID- 16350420 TI - The appearance of foramen in the internal aspect of the mental region of mandible from Japanese cadavers and dry skulls under macroscopic observation and three dimensional CT images. AB - The lingual canal with foramen displays different appearances on the internal surfaces of mandible as confirmed by macroscopic observation and computerized tomography (CT). The lingual canal was observed in the inside of mental region run to the outside of lingual foramen, which is extend internally from mandibular canal in right and left sides of the mandible in cadavers (13 sides out of 88 sides) and in dry skulls (43 out of 94 sides) examined. The spinal foramen connected with mental canal occurred at the midline of mandible in 6 cases (6 out of 47 cases) in dry skulls. In this small foramen, the inferior alveolar artery give some branches to the inside of mental region at the anterior mandible and which may be run pass through the lingual canal to the lingual foramen, where they emerge to enter the mylohyoid or anterior belly of digastric muscles. The observations of these are important considerations for surgical placement of dental implants in the region in the mandible. PMID- 16350421 TI - Morphological observation of the horseshoe kidney with special reference to the vascular system in 2 Japanese cadavers. AB - Two cases of the horseshoe kidney in Japanese cadaver were reported in this paper. The kidneys and their associated vessels in the retroperitoneal cavity were carefully examined, and the histological examination of the isthmus was performed. In Case 1, four arteries arose from the abdominal aorta. One right and two left renal arteries were distributed to the apical, upper, middle and posterior regions of the kidney, respectively, and the artery of isthmus entered the lower segments and the isthmus. In Case 2, six arteries arose from the abdominal aorta. Among three arteries arose from the inferior end of the aorta and entered the lower segments and the isthmus. Histological study revealed that the isthmuses consisted of collecting tubes, glomeruli and urinary tubules and fibrous connective tissue. The incidence of the horseshoe kidney during the dissecting practice at Tokyo Medical University in a period of 24 years from 1980 to 2003 was estimated to be 0.16% (2 out of the 1,219 cadavers). The anatomical and embryological significance of this anomaly and its associated vascular system were discussed. And the histology of the parenchymal structure of the isthmus in the horseshoe kidney containing either fibrous connective tissue or renal parenchyma was also analyzed in this report. PMID- 16350422 TI - Rare courses of the mandibular canal in the molar regions of the human mandible: a cadaveric study. AB - The inferior alveolar artery, vein and nerve send some branches to the molar teeth via the mandibular canal to the mental foramen. The present study attempted to define the presence and course of the mandibular canal in the mandible with the alveolar process by macroscopic cadaveric dissection and computerized tomography (CT) in order to provide information that might prevent injuries to vessels and nerves at risk during root canal treatment. We identified the position of the mandibular canal within a 30% ratio of the distance from inferior border of mandible to the apices of the root for 39 out of 131 sides (mesial root of first molar, 20%; distal root of first molar, 22.6%; mesial root of second molar, 27.8% and distal root of second molar, 47%) on panoramic X-ray observation. In one cadaver (male, 64 years old), the root apex of the second molar was in close proximity to the upper bony mandibular canal. Macroscopic dissection and computerized tomography showed that the main trunks of the inferior alveolar artery, vein, and nerve were in tight contact with the apex of the second molar. These observations of the anatomic course of the mandibular canal will be important to consider during root canal treatment of mandibular teeth. PMID- 16350424 TI - First Annual Chicago Supportive Oncology Conference: focusing on quality of life as essential to quality care. PMID- 16350423 TI - Staining of the Reinke crystalloids in the human testis--re-evaluation study. AB - Reinke crystalloid (CR)s are rod or corn shaped structures present in the interstitial cells (Leydig cell) of human testis. Due to existing controversies and because of their importance in histological studies, we decided to reevaluate their staining behavior with various dyes. Earlier it has been shown that hematoxylin and eosin dyes do not stain CRs even though their protein nature remains undisputed. In the present study, sections of testicular tissues embedded in glycol-methacrylate (GMA) showed that hematoxylin binds to CRs non specifically and eosin stains them specifically. The reasons for stainability and/or non stainability of CRs with hematoxylin, eosin and other dyes are discussed. PMID- 16350425 TI - Intrathecal drug delivery for the management of cancer pain: a multidisciplinary consensus of best clinical practices. AB - A substantial number of patients with cancer suffer considerable pain at some point during their disease, and approximately 25% of cancer patients die in pain. Providing effective pain management for patients with severe pain that impacts quality of life can present the oncologist or palliative care specialist with complex clinical challenges that often require multifaceted therapeutic measures. This paper presents multidisciplinary consensus-based recommendations for the treatment of intractable cancer pain using intrathecal drug delivery systems, which offer rapid and effective pain relief with less toxicity relative to oral or parenteral administration. Intrathecal drug delivery systems can be highly effective in a variety of patient settings, including cases of refractory pain, diminished performance status, poor tolerability of oral medications, polyanalgesia for complex pain, and inadequate dosing due to addiction concerns. The use of implantable or external systems is discussed, as well as implantation procedures, drug titration recommendations, and management of potential side effects. The authors offer a newly developed algorithm for delivering intraspinal analgesia in patients with cancer. The intent is that increased understanding of available options for truly effective pain management in the oncology and palliative care arena and the benefits of multidisciplinary cooperation will translate into genuine improvements in patient quality of life and a measurable decrease in the number of patients who suffer needlessly in their final days. PMID- 16350426 TI - Two decades of improved pain control. PMID- 16350427 TI - Neuropathic pain and balance problems are poorly evaluated in patients receiving taxane-based chemotherapy. PMID- 16350428 TI - Experimental L-glutamine agent shown effective in oral mucositis. PMID- 16350429 TI - A retrospective cohort study to assess the impact of therapeutic substitution of darbepoetin alfa for epoetin alfa in anemic patients with myelodysplastic syndrome. AB - Darbepoetin alfa and epoetin alfa are used to treat anemia in the undertreated population of patients with myelodysplastic syndrome (MDS). We implemented guidelines to switch anemic patients with MDS from epoetin alfa 40,000 U weekly to darbepoetin alfa 200 microg every 2 weeks and then conducted a retrospective cohort study of the initial 263 treated patients. Patients (> or = 18 years old, MDS diagnosis) were either previously treated with epoetin alfa (received 16 weeks of prior epoetin alfa and either switched to darbepoetin alfa or remained on epoetin alfa) or treatment-naive (no previous erythropoietin therapy and received only 1 agent for 16 weeks). Both major response and minor response based on the International Working Group criteria were calculated. The study was not powered to statistically compare treatment groups; values presented are for descriptive purposes only. Data from 244 patient records were included: 142 previous epoetin alfa patients (80 switched to darbepoetin alfa, 62 remained on epoetin alfa) and 102 naive patients (56 darbepoetin alfa, 46 epoetin alfa). Major response rates were similar between treatment groups in both the naive (46% for darbepoetin alfa, 35% for epoetin alfa) and previous epoetin alfa groups (26% for darbepoetin alfa, 17% for epoetin alfa). Overall response rates were 42%-76% across treatment groups. No differences in transfusions across groups were observed. Treatment of anemic patients with MDS with either darbepoetin alfa or epoetin alfa appeared to be effective. Whereas epoetin alfa was most frequently administered on a weekly basis, darbepoetin alfa was most frequently administered every 2 weeks, which may offer the benefit of convenience with its less frequent dosing. PMID- 16350430 TI - Palliative care for Muslim patients. PMID- 16350431 TI - CMS quality care demonstration project shows value despite methodological issues. PMID- 16350432 TI - Physical, psychological, and spiritual transformation at life's end: toward a theory of convergence. PMID- 16350433 TI - Causal connectivity of evolved neural networks during behavior. AB - To show how causal interactions in neural dynamics are modulated by behavior, it is valuable to analyze these interactions without perturbing or lesioning the neural mechanism. This paper proposes a method, based on a graph-theoretic extension of vector autoregressive modeling and 'Granger causality,' for characterizing causal interactions generated within intact neural mechanisms. This method, called 'causal connectivity analysis' is illustrated via model neural networks optimized for controlling target fixation in a simulated head-eye system, in which the structure of the environment can be experimentally varied. Causal connectivity analysis of this model yields novel insights into neural mechanisms underlying sensorimotor coordination. In contrast to networks supporting comparatively simple behavior, networks supporting rich adaptive behavior show a higher density of causal interactions, as well as a stronger causal flow from sensory inputs to motor outputs. They also show different arrangements of 'causal sources' and 'causal sinks': nodes that differentially affect, or are affected by, the remainder of the network. Finally, analysis of causal connectivity can predict the functional consequences of network lesions. These results suggest that causal connectivity analysis may have useful applications in the analysis of neural dynamics. PMID- 16350434 TI - Computational model of an adaptive rhythm generator within the olivocerebellar system. AB - The aim of this study is to investigate via computation whether the olivocerebellar system is capable of functioning as an adaptive rhythm generator. For this purpose, a detailed and physiologically realistic computational model of the olivocerebellar system is developed, based on the known intrinsic cell and network topological properties of this brain system. The present network, where individual cells are modelled by leaky integrate-and-fire units, converts the irregular spikes produced by the olivary cells into a precise rhythmic signal at the output. The simulation results reveal that the computational model, which normally does not exhibit any rhythmic activity, could be switched into a new mode in which it functions as a rhythm generator producing pulses within three different frequency ranges corresponding to alpha, beta, or gamma bands, respectively. In either mode of operation, the firing rates of all simulated cell types are observed to match real data. The results of this study therefore support the experimental findings of researchers who argue that a biological clock producing rhythmic pulses within different temporal ranges is located within the cerebellum. PMID- 16350435 TI - Optimal filtering for spike sorting of multi-site electrode recordings. AB - We derive an optimal linear filter, to reduce the distortions of the peak amplitudes of action potentials in extracellular multitrode recordings, which are due to background activity and overlapping spikes. This filter is being learned very efficiently from the raw recordings in an unsupervised manner and responds to the average waveform with an impulse of minimal width. The average waveform does not have to be known in advance, but is learned together with the optimal filter. The peak amplitude of a filtered waveform is a more reliable estimate for the amplitude of an action potential than the peak of the biphasic waveform and can improve the accuracy of the event detection and clustering procedures. We demonstrate a spike-sorting application, in which events are detected using the Mahalanobis distance in the N-dimensional space of filtered recordings as a distance measure, and the event amplitudes of the filtered recordings are clustered to assign events to individual units. This method is fast and robust, and we show its performance by applying it to real tetrode recordings of spontaneous activity in the visual cortex of an anaesthetized cat and to realistic artificial data derived therefrom. PMID- 16350436 TI - Acne, lasers, and light. PMID- 16350437 TI - Primary malignant tumors of the nail unit. PMID- 16350438 TI - Melanocytic nevi: challenging clinical situations in pediatric dermatology. PMID- 16350439 TI - Genomic analysis of melanocytic neoplasia. PMID- 16350440 TI - Dermatopathology update of "newer" dermatologic manifestations of systemic disease. PMID- 16350441 TI - High-risk cutaneous squamous cell carcinoma: identification and management. PMID- 16350442 TI - Neonatal violaceous skin lesions: expanding the differential of the "blueberry muffin baby". PMID- 16350443 TI - Practical review and recent developments in pediatric dermatopathology. PMID- 16350444 TI - Cutaneous appendage tumors: familial cylindromatosis and associated tumors update. PMID- 16350445 TI - Narrowband ultraviolet B therapy for psoriasis and other skin disorders. PMID- 16350446 TI - Chemokines and inflammatory skin diseases. PMID- 16350447 TI - New drugs, new rashes: update on cutaneous drug reactions. PMID- 16350448 TI - Advances in inherited epidermolysis bullosa. PMID- 16350449 TI - Epidermal stem cells. PMID- 16350450 TI - Antimicrobial peptides: effectors of innate immunity in the skin. AB - The ability of the cutaneous barrier to help defend the body against pathogens relies on both acquired and innate immune responses. Recently, a large body of research has suggested that a critical component of the innate immune response in the skin is 3 antimicrobial peptides: the cathelicidins, defensins, and dermcidins. These 3 classes of peptides have been shown to act as antimicrobials by directly inhibiting pathogen growth as well as potentiating other branches of the innate, humoral, and cell-mediated immune system. Here, we review the antimicrobial peptides with an emphasis on their role in the cutaneous immune response. We present an overview of defensin, cathelicidin, and dermcidin physiology, elucidating their various functions. In addition, we delve into the role of these peptides in specific dermatologic conditions including wound healing, atopy, and microbial infection. Finally, we discuss the future of antimicrobial peptide research including therapeutic options. PMID- 16350451 TI - Depolarisation phenomena in traumatic and ischaemic brain injury. AB - 1. Cortical spreading depression is a non-physiological global depolarisation of neurones and astrocytes that can be initiated with varying degrees of difficulty in the normally perfused cerebral cortex in the experimental laboratory. Induction is typically with electrical stimulation, needling of the cerebral cortex, or superfusion of isotonic or more concentrated potassium chloride solution. The phenomenon propagates across the cerebral cortex at a rate of 2-5 mm per minute, and is accompanied by marked but transient increases in cerebral blood flow, in local tissue oxygen tension, and most probably in metabolic rate. 2. Peri-infarct depolarisation is also a depolarisation event affecting neurones and glia, with an electrophysiological basis similar or identical to CSD, but occurring spontaneously in the ischaemic penumbra or boundary zone in focal cerebral cortical ischaemia. Most such events arise from the edge of the ischaemic core, and propagate throughout the penumbra, at a rate similar to that of cortical spreading depression. 3. Cortical spreading depression in the normally perfused cortex does not result in histological damage whereas peri infarct depolarisations augment neuronal damage in the penumbra, and are believed by many authors to constitute an important, or the principal, mechanism by which electrophysiological penumbra progressively deteriorates, ultimately undergoing terminal depolarisation and thus recruitment into an expanded core lesion. 4. There is some experimental evidence to suggest that under some circumstances induction of episodes of cortical spreading depression can confer protection against subsequent ischaemic insults. 5. Although cortical spreading depression and peri-infarct depolarisations have been extensively studied in the experimental in vivo models, there is now clear evidence that depolarisations also occur and propagate in the human brain in areas surrounding a focus of traumatic contusion. 6. Whether such events in the injured human brain represent cortical spreading depression or peri-infarct depolarisation is unclear. However, invasive and probably non-invasive monitoring methods are available which may serve to distinguish which event has occurred. 7. Much further work will be needed to examine the relationship of depolarisation events in the injured brain with outcome from cerebral ischaemia or head injury, to examine the factors which influence the frequency of depolarisation events, and to determine which depolarisation events in the human brain augment the injury and should be prevented. PMID- 16350452 TI - What is magnetoencephalography and why it is relevant to neurosurgery? AB - Magnetoencephalography (MEG) is a relatively novel technique that allows the study of the dynamic properties of cortical activity. The functional localization of brain sources of MEG signals depends on the models used and it always has a certain degree of uncertainty. Nevertheless, MEG can be very useful in assisting the neurosurgeon in planning and carrying out brain surgery in, or around, eloquent brain areas, and in epilepsy surgery in pharmaco-resistant patients. The following three areas of application of MEG in neurosurgery are reviewed: (i) Presurgical functional localization of somatomotor eloquent cortex; (ii) Presurgical evaluation of epileptic patients. (iii) Functional localization of speech relevant brain areas. The performance of MEG in comparison with EEG and fMRI is discussed. PMID- 16350453 TI - Basic and clinical aspects of olfaction. AB - Disturbances of olfaction are a common occurrence in many neurological and neurosurgical patients and their correct diagnosis might be helpful in management and enhancement of quality of life. However, olfaction is seldom checked in most neurosurgical units and the "smell bottles" are often either absent or out of date. This chapter reviews systematically recent advances in our understanding of the anatomy, physiology (olfactory coding) and measurement of olfactory function in the human. The causes and symptoms of smell disorders, risk of damage to the olfactory system by various surgical procedures and, finally, the natural history of recovery and treatment of smell disorders, for example after trauma, are discussed. PMID- 16350454 TI - Cranial venous outflow obstruction and pseudotumor Cerebri syndrome. AB - The pathophysiology of PTS including idiopathic intracranial hypertension or 'BIH', remains controversial. The older literature frequently referred to pathology in the cerebral venous drainage but more modern imaging techniques (CT and early MR) failed to reveal gross venous pathology. The role of impaired cranial venous outflow has recently been re-examined in the light of new methods of investigation (advanced MR venography and direct microcatheter venography with manometry) and of treatment (venous sinus stenting). Venous sinus obstruction in PTS is a more common factor in the pathogenesis of the condition than previously recognised. Venous obstruction may be primary, that is, it is the underlying aetiological factor in PTS. Venous sinus obstruction may also be secondary to raised CSF pressure which may exacerbate problems with intracranial compliance and raised CSF pressure. Early experience with venous stenting suggests that it may be a helpful treatment for patients with PTS but more experience and longer follow-up is required to define the subgroups of patients for whom it is most appropriate. PMID- 16350455 TI - Sacral neuromodulation in lower urinary tract dysfunction. AB - Vesico-urethral dysfunction is a major problem in daily medical practice due to its psychological disturbances, its social costs and its high impact on quality of life. Recently, sacral neuromodulation, namely the electrical stimulation of the sacral nerves, appears to have become an alternative for radical bladder surgery particularly in cases of idiopathic bladder overactivity. The mechanism of action is only partially understood but it seems to involve a modulation in the spinal cord due to stimulation of inhibitory interneurons. Temporary sacral nerve stimulation is the first step. It comprises the temporary application of neuromodulation as a diagnostic test to determine the best location for the implant and to control the integrity of the sacral root. If test stimulation is successful, a permanent device is implanted. This procedure is safe in experienced hands. So-called idiopathic bladder overactivity still the major indication for this technique. Patients not likely to benefit from the procedure were those with complete or almost complete spinal lesions, but incomplete spinal lesions seemed to be a potential indication. This technique is now also indicated in the case of idiopathic chronic retention and chronic pelvic pain syndrome. When selection is performed, more than three-quarters of the patients showed a clinically significant response with 50% or more reduction in the frequency of incontinent episodes, but the results vary according to the author's mode of evaluation. From the economic point of view, the initial investment in the device is amortized in the mid-term by savings related to lower urinary tract dysfunction. Finally, this technique requires an attentive follow-up and adjustments to the electric parameters so as to optimize the equilibrium between the neurological systems. PMID- 16350456 TI - Prevention and treatment of postoperative pain with particular reference to children. AB - Pain therapy is an important aspect of medical practice for patients of all ages, to optimize care, to obtain an adequate quality of life and to improve their general conditions. Pain is among the most prevalent symptoms experienced by patients undergoing surgery. The success of postoperative pain therapy depends on the ability of the clinician to assess the presenting problems, identify and evaluate pain syndromes and formulate a plan for comprehensive continuing care. The prevalence of acute pain has led to the need to develop techniques for the assessment and management of this symptom in order to focus the attention on an interdisciplinary therapeutic approach (including pharmacologic, cognitive behavioral, psychologic and physical treatment) and on the timing of different interventions (pre and postoperative). In this chapter we describe the principal therapeutic approaches to control pain in post-operative patients, such as non opioid, opioid and adjuvant analgesics with particular attention in paediatric age. Moreover we report the principal scales to assess the pain intensity in the post-operative period. The need of a multidisciplinatory team and of a pre and postoperative pain management program represents an important goal in order to obtain effective pain relief and optimize pediatric care and rapid recovery. The introduction of a perioperative team service will improve the approach to pain management programs and it is considered the most important challenge for future. PMID- 16350457 TI - Revelation. PMID- 16350458 TI - The 2005 Gold Medal for Distinguished Service--Dr. Robert Anderton. PMID- 16350459 TI - Guidelines for Infection Control in Dental Health Care Settings--a review. PMID- 16350460 TI - Personnel health elements of infection control in the dental health care setting- a review. PMID- 16350461 TI - Chemical disinfectants in dental practice--a review. PMID- 16350462 TI - Methods of dental instrument processing, sterilization, and storage--a review. AB - A comprehensive instrument processing and sterilization program in the dental office is essential to ensure that the DHCP and the public are protected from disease transmission due to contaminated instruments/ devices. The Centers for Disease Control and Prevention and other organizations have made recommendations to help dental personnel with this aspect of patient care. By following the CDC's latest guidelines, the DHCP can develop an optimal program of dental instrument processing, sterilization and storage. PMID- 16350463 TI - Dental water quality--the scientific evidence for current CDC guidelines. AB - Newly revised and updated guidelines from the Centers for Disease Control and Prevention recommend adoption of procedures to improve the microbiological quality of water used in dental treatment. These recommendations represent the consensus of experts from a wide range of scientific and clinical disciplines and are based on the best currently available evidence. Dentists and the manufacturers of dental equipment must consider the infection control, occupational health and risk-management implications of biofilm colonization in dental equipment, and take appropriate measures to provide water of appropriate microbiological quality clinical procedures. PMID- 16350464 TI - Why dentists should plan for long term care. PMID- 16350465 TI - Oral and maxillofacial pathology case of the month. Lymphangioma (lymphatic malformation). PMID- 16350466 TI - Over a barrel: corporate corruption of science and its effects on workers and the environment. AB - Although occupational and environmental diseases are often viewed as isolated and unique failures of science, the government, or industry to protect the best interest of the public, they are in fact an outcome of a pervasive system of corporate priority setting, decision making, and influence. This system produces disease because political, economic, regulatory and ideological norms prioritize values of wealth and profit over human health and environmental well-being. Science is a key part of this system; there is a substantial tradition of manipulation of evidence, data, and analysis, ultimately designed to maintain favorable conditions for industry at both material and ideological levels. This issue offers examples of how corporations influence science, shows the effects that influence has on environmental and occupational health, and provides evidence of a systemic problem. PMID- 16350467 TI - Maximizing profit and endangering health: corporate strategies to avoid litigation and regulation. AB - Corporations and industries use various tactics to obscure the fact that their products are dangerous or deadly. Their aim is to secure the least restrictive possible regulatory environment and avert legal liability for deaths or injuries in order to maximize profit. They work with attorneys and public relations professionals, using scientists, science advisory boards; front groups, industry organizations, think tanks, and the media to influence scientific and popular opinion of the risks of their products or processes. The strategy, which depends on corrupt science, profits corporations at the expense of public health. Public health professionals can learn from this strategy how to effectively build scientific and public opinion that prioritizes both good science and the public health. PMID- 16350468 TI - Lifting the veil of secrecy from industry funding of nonprofit health organizations. AB - Those who provide information about scientific issues and science policy normally present themselves as being objective and "scientific." This article describes a range of health charities, professional associations, nonprofit advocacy organizations, and industry-created organizations that receive significant funding from industry. In some cases, industry appears either to influence an organization's positions or to limit an organization's freedom to speak out on matters of interest to the funders. Nonprofit organizations need to consider the potential influence on their independence if they accept funding from interested companies and trade associations. PMID- 16350469 TI - Business bias: how epidemiologic studies may underestimate or fail to detect increased risks of cancer and other diseases. AB - In spite of claiming primary prevention as their aim, studies of potential occupational and environmental health hazards that are funded either directly or indirectly by industry are likely to have negative results. The authors present three common scenarios in which faulty design of epidemiologic studies skews results, and list 15 study design flaws that lead to results that are dangerously misleading with regard to both the evaluation and the improvement of public health. PMID- 16350470 TI - Abuse of epidemiology: automobile manufacturers manufacture a defense to asbestos liability. AB - Much of the "debate" about the relationship between asbestos exposure from automobile brake work and asbestos-induced cancer has been fueled by studies that have been funded by corporations with billions at stake in tort litigation. The authors explore how asbestos-lined brake manufacturers have corrupted medical literature to escape liability, analyzing studies funded by these companies to enable them to claim that work with asbestos brake linings never causes mesothelioma. They reveal how the companies have redefined scientific criteria for the determination of cause-effect relationships and manipulated scientific data to give the impression of an absence of effect. But the absence of evidence is not evidence of the absence of an effect. PMID- 16350471 TI - Safeguarding scientific evaluations by governmental agencies: case study of OSHA and the 1,3-butadiene classification. AB - Using the example of the industry pressures on OSHA that preceded the Agency's downgrading the carcinogenic potential of butadiene from "human carcinogen" to "possibly carcinogenic to humans" in the face of scientific evidence, the author warns of the danger to public health of the infringement on governmental agencies' decision making by special-interest groups. PMID- 16350472 TI - Industry efforts to weaken the EPA's classification of the carcinogenicity of 1,3 butadiene. AB - Industry's success in negotiating away science at the level of governmental regulatory agencies is eroding the integrity of the regulatory process. Maneuvers used by industry to weaken the U.S. Environmental Protection Agency's classification of the carcinogenicity of butadiene are described. PMID- 16350473 TI - Ethyl-leaded gasoline: how a classic occupational disease became an international public health disaster. AB - The author describes the controversy about the use of tetraethyl lead (TEL) as a gasoline additive. Early warnings were ignored by industry, and as leaded gasoline became more profitable, scientists willing to support industry were financed as guardians of the scientific criteria for lead's health impacts. Controversy erupted in 1924 after refinery accidents left workers dying from violent insanity. In efforts to protect their profits, industry executives falsely claimed there was no alternative to leaded gasoline. Fifty years passed before scientific, court, and regulatory challenges had any influence. When independent research finally emerged, the results were damning enough to support an international phase-out of leaded gasoline. PMID- 16350474 TI - Mining and mendacity, or how to keep a toxic product in the marketplace. AB - The corruption of medical evidence about the hazards of asbestos began with the Canadian mines. Quebec at one time boasted ten of the 13 mines in Canada. Work conditions in the mines were harsh, and the mills were full of airborne fiber. Given the size of the industry, the Quebec mines were where occupational asbestosis should first have been identified, but research at the mines was done in company towns, where clinics were staffed by company doctors. Since the late 1920s U.S. parent companies and their Canadian subsidiaries have maintained that there is little if any disease among mine workers. Asbestosis in textile workers, they have claimed, has been due to the conditions in that industry and not the inherent dangers of asbestos. That fiction continues to shape the discourse about the usefulness of asbestos. PMID- 16350475 TI - Fluoride poisoning: a puzzle with hidden pieces. AB - Key industry data regarding harm from chronically inhaled fluoride have been unavailable publicly for decades. Recent unveiling of unpublished reports reveals three examples of data mishandling that disguised the need for more stringent occupational standards for particulate and gaseous fluorides and fluorine. Injury reports from workers handling chemicals show that unjustifiable reductions of injury and disability numbers in the process of publication shifted concern from respiratory to mineralized tissue damage. Selective editing and data omissions allowed bias that fluoride reduces caries without detrimental effects. Finally, industry's failure to publish an important industry-funded laboratory study buried knowledge of low thresholds for fluoride-induced lung disease. Data from that study are presented to clarify the dose- and duration-dependent changes caused by chronic inhalation of calcium fluoride. PMID- 16350477 TI - Genetic engineering in agriculture and corporate engineering in public debate: risk, public relations, and public debate over genetically modified crops. AB - Corporations have long influenced environmental and occupational health in agriculture, doing a great deal of damage, making substantial profits, and shaping public debate to make it appear that environmental misfortunes are accidents of an otherwise well-functioning system, rather than systemic. The debate over the genetically modified (GM) crops is an example. The largest producer of commercial GM seeds, Monsanto, exemplifies the industry's strategies: the invocation of poor people as beneficiaries, characterization of opposition as technophobic or anti-progress, and portrayal of their products as environmentally beneficial in the absence of or despite the evidence. This strategy is endemic to contemporary market capitalism, with its incentives to companies to externalize health and environmental costs to increase profits. PMID- 16350476 TI - The dirty work of promoting "recycling" of America's sewage sludge. AB - Serious illnesses, including deaths, and adverse environmental impacts have been linked to land application of sewage sludge. EPA and the wastewater treatment industry have worked with Congress to fund wastewater trade associations to promote land application, supporting industry-friendly scientists and discouraging independent research, to prevent local governments from restricting land application and to thwart litigation against municipalities and the industry. PMID- 16350478 TI - Who's afraid of national laws? Pesticide corporations use trade negotiations to avoid bans and undercut public health protections in Central America. AB - The agrochemical industry is using trade agreements to block proposed bans on pesticides identified as the worst occupational health hazards by a multi-country illness surveillance program in Central America. Through privileged access to closed-door negotiations, industry inserted deregulatory mechanisms, including a regional pesticide registry that invalidates national laws, investors' rights protection, and increased intellectual property protections, into the draft Central American Customs Union and the Central American Free Trade Agreement. These agreements undermine health-based national pesticide registration requirements; weaken health ministries' role in pesticide control; block marketing of cheaper, less toxic pesticides; and have a chilling effect on future pesticide regulatory activity. So long as corporations have privileged access to the trade negotiations and civil society is excluded, the resulting agreements will benefit special interests at the expense of public health. PMID- 16350479 TI - A systemic approach to occupational and environmental health. AB - As the corporate role in occupational and public health receives increased scrutiny, it is essential to recognize that it is not sufficient to identify specific acts of malfeasance or influence, or even to campaign to address them. A more comprehensive and systemic framework for understanding the role of corporations requires consideration of corporate power and its effects as endemic features of national socioeconomic systems and the rapidly integrating global order. The underlying social structures that produce social and environmental problems, and undermine reform, make systemic change necessary. Identifying this "structure of harm" provides important implications for researchers, policymakers, activists, and others trying to address environmental and social problems, particularly with regard to integrating efforts to address immediate impacts with those for longer-term, systemic change. PMID- 16350480 TI - Chevron-Texaco's science. PMID- 16350481 TI - Circadian variation and outcome of in-hospital cardiopulmonary resuscitation. AB - OBJECTIVE: Assessment of contemporary in-hospital cardiopulmonary resuscitation outcomes. METHODS: The study was designed as a retrospective, single-institution, registry investigation of all adult patients for whom a cardiopulmonary resuscitation (CPR) was attempted between October 1, 2002 and September 30, 2004. Hospital, patient's, arrest and outcome variables were included based on the "Utstein Style" of reviewing, reporting, and conducting research on in-hospital resuscitation. RESULTS: 202 patients (94 women, age 70 +/- 15 years) received in hospital CPR during the study period. One hundred-two events were primary respiratory arrests. Overall, 67% (95% CI 59%-72%) of patients were successfully resuscitated and 38% (95% CI 31%-45%) survived to hospital discharge. Of the patients with initial respiratory depression followed by circulatory collapse, 57% (95% CI 47%-66%) were alive at the end of hospitalization, while 20% (95% CI 13.3%-28.8%) of patients with cardiac arrest survived to hospital discharge (OR=5.2, 95% CI=2.8-9.5, P<0.0001). A circadian variation in the occurrence of primary cardiac arrests was noted with a nadir between midnight and 0600 hours. Though there were fewer events during overnight hours, CPR for patients with a primary cardiac arrest occurring at this time was less likely to be successful (OR = 2.5, 95% CI=1.01-6.1, p=0.04). This did not result in a difference in survival to discharge (OR=2.6, 95% CI=0.7-9.4, p=0.14). CONCLUSION: Primary cardiac arrest has a significantly worse in-hospital outcome when compared to an initial respiratory event. The finding of a circadian variation in both the incidence and outcome of in-hospital primary cardiac arrest is intriguing and warrants further study. PMID- 16350482 TI - Critical care physicians' practices and attitudes and applicable statutes regarding withdrawal of life-sustaining therapies. AB - OBJECTIVE: To describe intensivist-educators' practices and opinions regarding the withdrawal of life-sustaining therapies, and to juxtapose these with applicable end-of-life statutes. METHODS: A questionnaire was sent to critical care program directors in Connecticut, New York, Illinois, California, Florida and Texas (to examine regional variability). Statutes regarding end-of-life care were ascertained for these states. RESULTS: Sixty-eight of 128 queried intensivists responded to the questionnaire. They reported having withdrawn life sustaining therapies to a median of 28 patients in the previous year. Overall, respondents answered that of patients in whom they had withdrawn life-sustaining therapies, 55% were terminal, 20.5% weren't necessarily terminal but advance directives were applied to the situation and 22% weren't necessarily terminal but families requested withdrawal. The ranges of responses were highly variable (ranging from 5%-100% patients deemed terminal). All respondents agreed that withdrawal should be allowed in non-terminal conditions in which the likelihood or quality of survival would be unacceptable to the patient. Forty-eight respondents answered that advanced Alzheimer's Disease qualifies as a terminal illness. Connecticut's end-of-life statutes were unlike those in the five other states that allowed withdrawal of life-sustaining therapies based on advance directives and/or surrogates' consent. Practices of Connecticut respondents were similar to other sampled states. CONCLUSIONS: These data suggest that Connecticut's statutes requiring that patients be deemed terminal (or permanently unconscious) before withdrawal of care are dissimilar from those in several other states. Connecticut intensivists who responded to our survey appear to practice end-of-life care that is similar to practices elsewhere. PMID- 16350483 TI - Necrobiosis lipoidica resulting in squamous cell carcinoma. AB - Necrobiosis lipoidica is a chronic degenerative disease of dermal connective tissue, of unknown etiology, which occurs mostly in diabetic patients. It may occasionally be complicated by squamous cell carcinoma. Since this combination is rare, a high index of suspicion is necessary to establish the diagnosis. We present a case and a review of the literature. PMID- 16350484 TI - Electrocardiographic manifestations of 5-fluorouracil cardiotoxicity. PMID- 16350485 TI - The Conscientious Practice Policy: a futility policy for acute care hospitals. AB - Much attention has been paid in recent years to the conflict that may occur when patients or their families insist on a therapy that the physician feels would be futile. In 1999 the Council on Ethical and Judicial Affairs of the American Medical Association recommended that all health-care institutions adopt a policy on medical futility that follows a fair process. Development of such a policy has proved problematic for many hospitals. The Conscientious Practice Policy at Lawrence & Memorial Hospital was developed as a response to the AMA recommendation. It outlines a specific process to be followed in the event that a physician wishes to refuse to provide a requested therapy, whether that refusal is based on perceived futility or other concerns. The policy was subsequently modified slightly and adopted by two other Connecticut acute care hospitals. PMID- 16350486 TI - Who is a doctor? PMID- 16350487 TI - Bell's palsy treated with cortisone. 1955. PMID- 16350488 TI - A new direction in health insurance. 1955. PMID- 16350489 TI - Health care for the uninsured: here we go again. PMID- 16350490 TI - A vulnerable population. PMID- 16350491 TI - 20,000 unaware they have HIV. PMID- 16350492 TI - Health workers in Africa need greater access to AIDS services. PMID- 16350493 TI - It's just between you and me ... and social services and the police. PMID- 16350494 TI - A wee problem. AB - Stress incontinence plagues many women's lives. It is more common than diabetes or asthma, and can have major social and financial implications. Although treatment is available, new research suggests that women are often being given inappropriate drugs. PMID- 16350495 TI - Family matters. AB - Helping relatives cope with the stress and practical difficulties of having a loved one in intensive care is the remit of senior nurses at James Cook University Hospital. Senior intensive care nurses take on the role of relatives' liaison nurse for two months at a time. They are entirely focused on the needs of relatives, having no clinical responsibilities. One of the most common needs relatives have is for temporary accommodation, as many live outside the area. The nurses accompany relatives when they speak to doctors so that they can reinforce information and explain further. PMID- 16350496 TI - Can you read the emotional radar? AB - Emotional intelligence means that your instinct picks up what is happening to those around you. And it is a skill that can be learned, provided you have the time and inclination. PMID- 16350497 TI - Do mum and dad know best? PMID- 16350498 TI - Taking loss in hand. PMID- 16350499 TI - The gift of sight. PMID- 16350500 TI - Charitable view. PMID- 16350501 TI - Smoking cessation for clients who are HIV-positive. AB - AIM: To identify smoking prevalence and behaviour in clients who are human immunodeficiency virus (HIV)-positive and to enrol those wanting to stop smoking in a smoking cessation programme. METHOD: A questionnaire consisting of 28 questions on smoking behaviour was developed. Over an eight-week period at the Immunology Clinic, Royal Prince Alfred Hospital, New South Wales, Australia, 68 clients were asked to complete the questionnaire; 23 were ineligible because they were non-smokers. Of the 45 respondents who completed the questionnaire, 39 were current smokers and six were ex-smokers. Twenty seven clients enrolled in the smoking cessation programme. RESULTS: Smoking behaviour did not change due to a diagnosis of HIV or living with HIV. Stress was the main reason for recommencing smoking. At the end of the programme, 22 per cent (n = 6) had ceased smoking and 40 per cent (n = 11) had reduced nicotine intake. CONCLUSION: Smoking behaviour is complex and the reasons clients continue to smoke are multifactorial. Therefore, smoking cessation programmes should reflect this complexity. PMID- 16350502 TI - The shadowing experience for nursing students. AB - The shadowing scheme supports nursing students in learning how to manage and identify the characteristics of good leadership. Derby Hospitals NHS Foundation Trust and the University of Nottingham offered nursing students the opportunity to shadow service leaders and managers during their final semester. This initiative promotes an effective service, takes account of students' needs and interests and supports the government's modernisation agenda and the NHS Plan (Department of Health (DH) 2000). PMID- 16350503 TI - Leadership and support for safety. AB - This is the second in a series of seven articles on the National Patient Safety Agency's Seven Steps to Patient Safety (NPSA 2004a). Leadership and support for staff can be achieved by ensuring that patient safety has a clear focus throughout the organisation. This article considers the context in which nurses work and provides examples of what can be done at organisational, directorate and team levels to ensure patient safety. PMID- 16350504 TI - Wound bed preparation and the TIME principles. AB - The principles of wound bed preparation in the management of chronic wounds are described using the Tissue, Infection, Moisture, Edge (TIME) framework. TIME offers a systematic approach to wound healing, which involves eliminating non viable tissue, controlling infection, restoring moisture balance and promoting epithelial advancement. The application of each of these principles to wound management in the clinical setting is discussed. PMID- 16350505 TI - Surgical nursing care. PMID- 16350506 TI - Vital statistics. PMID- 16350507 TI - Reach out to a world of learning. PMID- 16350509 TI - Are medical dramas a public health hazard? PMID- 16350508 TI - RMNs still await rules on restraint. PMID- 16350510 TI - Pension penalties. PMID- 16350511 TI - Leading the way in mother's care. PMID- 16350512 TI - How to maintain optimal glycaemic control in diabetes. AB - Despite national and local guidelines for glycaemic control, current management of glycaemia can fall significantly short of accepted goals. This article discusses the recommended glycaemic control goals and barriers that can prevent their achievement. It examines key recommendations to enable health care providers to overcome these barriers. PMID- 16350513 TI - Appendicitis. PMID- 16350514 TI - The liver part l: anatomy. AB - Understanding the anatomy of the hepatobiliary system provides the basis for the holistic care of patients with a liver disorder. In the second part of this series of articles, the functions of the liver will be examined. PMID- 16350515 TI - Nutrition and hydration tips for stroke patients with dysphagia. AB - This article examines the nursing care that should be given to patients who have suffered a stroke (cerebrovascular accident or CVA) that has caused them to have dysphagia. It focuses on the maintenance of their nutrition and hydration needs through safe oral feeding practices and how the risk of aspiration can be managed. This is an important nursing issue, particularly as the Royal College of Physicians of Edinburgh (RCP, 2000) states that nutrition and hydration needs are often not met. PMID- 16350516 TI - Reducing overcrowding on student practice placements. AB - The UK is facing a major shortage of registered nurses. One government strategy for addressing this shortage has been the nationwide increase in the nursing student population. Large student cohorts have resulted in placements being overcrowded with students and, as a consequence, the quality of learning experiences has diminished. A mental health NHS trust in London in partnership with local higher education institutes has created a framework to relieve the pressure on placements. This article discusses the implementation of the framework and outlines the results, challenges and recommendations. PMID- 16350517 TI - Zidovudine. PMID- 16350518 TI - Defining nursing knowledge. AB - The question of what constitutes nursing knowledge is a vital one for nurses to consider for a number of reasons. Nurses use a wide range of knowledge in practice, some theoretical and some practical, so identifying what nursing knowledge is should be central to practice. This paper attempts to define nursing knowledge by discussing the evidence. It suggests that such knowledge is important to raise awareness of personal and professional accountability, inform the dilemmas of practice and improve patient care. PMID- 16350519 TI - Needs-led management in the community. PMID- 16350520 TI - A checklist for planning flu vaccination clinics for 2006. AB - Flu vaccination clinics have reached crisis point this year, with increasing numbers of patients and a shortage of vaccine. Joyce Skeet provides a checklist to help in the planning of the 2006 programme. PMID- 16350521 TI - Advice on healthy eating for older people. AB - As part of its Food and Health Action Plan, the Department of Health is working with the food industry, and with other stakeholders, to establish a coherent national plan to help people in England improve their diets. Maintaining a healthy diet is important for all age groups, but healthy older people have particular needs. Karen Fisher describes the specific nutritional issues affecting healthy older people and suggests advice that nurses can offer people during opportunistic consultations in primary care. PMID- 16350523 TI - Are nurses ready to take on the BNF? PMID- 16350522 TI - Helping people to stop smoking: the role of treatment products. AB - Advising patients to stop smoking is an important intervention that will have an effect on their immediate and future health. Nurses need to be familiar with the numerous products that are available to assist smokers to give up cigarettes. Jennifer Percival describes how nicotine replacement therapy and bupropion can help smokers achieve success. PMID- 16350524 TI - Improving support for renal patients. PMID- 16350525 TI - Nursing wherever they are needed. PMID- 16350526 TI - Methods and models so well suited to general medicine simply do not fit mental health and never will. PMID- 16350527 TI - [Premorbid personality and aggressive behavior in residents of psychogeriatic nursing homes]. AB - In the present study, the correlation was examined between pre-morbid personality traits and the occurrence of aggressive behaviour in residents of a psychogeriatric nursing home. The participants in the study were the close relatives of 39 residents of a psychogeriatric nursing home. These residents were selected on the basis of two subscales of the BOP assessment scale for elderly patients: Aggressive and annoying behaviour, and mental disability. Residents were divided into two groups: one group did not display aggressive behaviour, the other group displayed a great deal of such behaviour. The groups were comparable with regard to the degree of mental disability. To ascertain pre-morbid personality traits, use was made of the hetero-anamnestic personality questionnaire (HAP) and the Quick Big Five (QBF). We found no significant differences in any of the premorbid personality traits between the two groups. We could not confirm the conclusion of other authors that emotional lability (neuroticism) is a personality trait which makes people susceptible to developing aggressive behaviour. In conclusion, our study did not confirm the hypothesis that pre-morbid personality traits are useful to explain the occurrence of aggressive behaviour in residents of a psychogeriatric nursing home. PMID- 16350528 TI - [Searching for rest in one's soul: the experience of mental health complaints in older Moroccan immigrants]. AB - The number of Moroccan elderly in the Netherlands is growing. Although many have mental health problems, the mental health services have difficulties in reaching them. This study reports on the experience of mental health problems of elderly Moroccan migrants, their search for help and their view on the role of the mental health services. Twenty-two Moroccan elderly (11 men, 11 women) aged 55-75 were interviewed with a semi-structured questionnaire. From the interviews and observations it appeared that these elderly experience their health as bad. When asked, they deny mental health problems. Stress and dejection are not experienced as relating to mental health but as part of life associated with ageing, living in a foreign country, having physical, social and financial problems. The elderly feel that God sets them these tasks to overcome by themselves. Achieving rest in one's soul is central in this process. There is a taboo on serious mental health problems and people are ashamed of them. Only people who neglect religious prescriptions can to be affected by mental health problems. The elderly Moroccans hardly consult mental health services for these problems because then it would become clear that they cannot overcome their problems by themselves. Furthermore, the mental health services are unknown. Therefore, those services have to pay attention to the specific experience of mental complaints of elderly Moroccans. By offering psycho-education the mental health services may come within reach of those elderly. PMID- 16350529 TI - [Vaccination of the elderly: an update]. AB - Vaccination of the elderly still requires attention. The vaccination coverage for tetanus, influenza and pneumococcal infections is merely 40, 60 and 30%, respectively. Besides a reduction in mortality (67%) and a reduction of hospitalisation for pneumonia and influenza (50%), vaccination against influenza also results in a decrease in cardio- and cerebrovascular morbidity (20%) as well as in a decrease in the frequency of doctor visits for respiratory infections for COPD patients. Vaccination of children and health care personnel can further reduce transmission of influenza and subsequent influenza related complications in the elderly. Pneumococcal invasive disease can be reduced by 50% through vaccination. Vaccination of children with the conjugate vaccine can further reduce the incidence of pneumococcal invasive disease in the elderly. Further improvements in vaccine coverage levels are needed, mainly among elderly persons, children and persons at increased risk. PMID- 16350530 TI - [Acute infectious (not Clostridium difficile-associated) diarrhea in the elderly]. AB - Acute diarrhoea, non-antibiotic associated, is a common problem and a significant cause of morbidity and mortality in old age. In most cases diarrhoea has an infectious etiology. A number of different micro organisms can cause infectious diarrhoea. Most frequent are viral infections with a benign evolution. Rehydration is the only important therapeutic measure. Infections with bacteria are less common, antibiotics should be prescribed only in severe cases, and when there is suspicion of invasive infections by enteropathogenic bacteria. PMID- 16350531 TI - [The biology of the Ixodes ricinus tick]. AB - More than 800 tick species have been reported world-wide however only about 30 tick species feed on humans, among them Ixodes ricinus, which is the most frequent tick species biting humans in Europe. It is the vector of Borrelia burgdorferi, which causes Lyme disease and of the tick-borne encephalitis virus. I. ricinus ticks pass through three developmental stages: larvae, nymphs and adults (females and males). More than 300 animal species have been reported as natural hosts for I. ricinus and 50 vertebrate species have been identified as reservoir hosts for B. burgdorferi. The density of this tick species may be very high, reaching in some places more than 300 ticks/100 m2. Attached ticks should be removed without delay with a blunt forceps. PMID- 16350532 TI - [Protection against insects]. AB - Successful protection against haematophagous insects and ticks, especially in areas where transmission of diseases occurs, requires a consistent application of a combination of appropriate measures. However, this can never substitute a chemoprophylaxis. Which measures have to be used depends on the circumstances under which they have to work. Indoor, physical means such as mosquito-screens on doors and windows, air-conditioners, and bed nets can be used to keep the insects away. These measures can be supplemented or supported by insecticides used as knock-down sprays, by electrical evaporation or for the treatment of screens and bed nets. In the field, if it is not possible to avoid mosquito-areas during phases of activity, appropriate clothing and repellents must provide the protection. Bright, wide pants and shirts of dense weaving covering as much skin as bearable should be preferred. Repellents are sprays, lotions, milks or creams which are evenly applied to the skin to prevent insects from biting. They contain synthetic or natural active substances of substantially varying effectiveness. The gold standard since about 60 years is diethylbenzamine (DEET). There are a few other active substances with a lower risk of side effects, however, combined with a lower effectiveness mainly on people with a high attractiveness for mosquitoes. Products containing an extract of Eucalyptus citriodora provide the best protection amongst those with natural active substances. Wearing bracelets or necklaces treated with repellents, acoustic devices (buzzers), electrocuters, topical or systemic Vitamin B1 or eating garlic are useless measures to prevent insects from biting. PMID- 16350533 TI - [Epidemiology of tick-borne encephalitis (TBE) in Switzerland 1984 to 2004]. AB - From 1984 to 2004 a total of 1370 TBE cases have been reported to the Federal Office of Public Health through the notification system. A significant increase of cases was observed in the nineties. Since 1999 the yearly average of reported cases was 101, corresponding to a yearly incidence of 1.4/10(5) inhabitants for all of Switzerland or to an incidence of approximately 6/10(5) if only the population living in endemic regions is taken into account. Male cases were twice as frequent as female ones. The age distribution showed two peaks in the age ranges of 6 to 14 and 60 to 69 years. Cases in children under 6 years of age were very rare with a yearly average of two hospitalised children. The severity of the disease increased with age. In 86% of the patients CNS involvement was present while 14% showed only flu-like symptoms. Ninety percent of patient with CNS involvement were hospitalised, 1,3% died (on average one death per year since 1999). Cases in fully vaccinated persons were very rare. The majority of patients had most probably been infected at their place of residence or in the nearer surroundings. The incidence in the large cities was low (< or = 1/10(5)/year). In smaller towns located directly in an endemic region, the incidence rose to as much as 18/10(5)/year. During the 21 years of observation, the endemic regions in four cantons known in the early eighties had clearly expanded and new infected regions appeared in seven other cantons. A frequent recreational or occupational exposure in wooded areas was mentioned by 85% of the patients. Although 80% of these concerned leisure time activities (mainly walking), the incidence is most probably still higher in occupationally exposed persons. This indicates that not only persons with leisure time exposure but also occupationally exposed persons are still insufficiently vaccinated to achieve an optimal control of this disease in Switzerland. PMID- 16350534 TI - [Prevention of tick-borne encephalitis (TBE)]. AB - Active immunization is the only reliable means of preventing TBE. Evidence suggests that effectiveness is greater than 95%. Breakthrough infections in fully immunized individuals, however, do occur. Recent data indicate that the duration of protection following basic immunization is substantially longer than previously appreciated. The vast majority of vaccinees are still seropositive > or = 8 years after the last dose. Thus, the current practice of administering booster doses every 3 years is a topic of intense debate. Until it is resolved, patients can be offered the determination of a serum anti-TBE IgG titre as an alternative to "blind" administration of a booster dose. It has recently been shown that seropositivity determined by some commonly used commercial ELISA tests correlates closely with the presence of protective antibody. In Switzerland, TBE vaccination is currently recommended for individuals older than 6 years of age who frequently dwell in endemic areas. Since 2005, health care insurances are required to cover for costs incurred by immunization according to these recommendations. PMID- 16350535 TI - [Lyme borreliosis--an overview]. AB - North American Lyme disease and European borreliosis differ in many aspects: Whereas in North America infection by Borrelia burgdorferi sensu stricto almost invariably leads to erythema migrans, frequently followed by oligoarthritis in untreated individuals, most infections in Europe are caused by Borrelia garinii and Borrelia afzelii and are contracted asymptomatically. A minority of the infected individuals develop neuroborreliosis, consisting of various combinations of meningitis, radiculitis and (cranial) neuritis. Lyme carditis, presenting with second or third degree atrioventricular block, is infrequent on both continents. Diagnostic limitations and the protean manifestations of Lyme borreliosis frequently give rise to misunderstandings and overdiagnosis of Lyme disease. Targeted use of Lyme serology, judicious application of a case definition, strict adherence to established treatment options and thorough patient information may lead the narrow path between unjustified exclusion and careless assumption of the diagnosis of Lyme borreliosis. PMID- 16350536 TI - [Diagnostic tests of Lyme borreliosis]. AB - The diversity of Borrelia burgdorferi subspecies (sensu lato) in Europe does not allow to simply adopt the US definitions and tests which were established for B. burgdorferi sensu stricto. Routine detection of Borrelia-specific antibodies now includes specific antigens of the subspecies B. afzelii and B. garinii. Thus, the 2-step serology of screening by ELISA and confirmation by immunoblot remains the most important diagnostic test of lyme borreliosis. The detection of borrelia by PCR or culture is of lesser importance due to the limited sensitivity, despite of the high specificity. In clinical practice, the risk of exposure to ticks and the stage of the clinical presentation guide the choice of appropriate diagnostic procedures and their interpretation. PMID- 16350537 TI - [Late manifestations of Lyme borreliosis]. AB - Month to years after an early local or an early disseminated infection some patients develop late manifestations of lyme borreliosis. Most frequently involved organs are the skin (acrodermatitis chronica atrophicans), joints (Lyme arthritis) and the nervous system. A history of exposure and the clinical picture may suggest Lyme borreliosis, however, confirmation by serological and other tests is needed. Antibiotic treatment during early stages normally prevents development of late manifestations. Late stages persist if not treated. By adequate antimicrobial therapy they are treatable and usually show a good prognosis. Recovery may be delayed, some patients suffer from residual difficulties. Currently there is no accepted case definition for a "post lyme syndrome". The term "chronic Lyme disease" suggests (a never proven) persistent infection by viable bacteria. Repeated and prolonged antibiotic treatments are not indicated. PMID- 16350538 TI - [Lyme borreliosis--treatment and prevention]. AB - Several antimicrobials are effective for the treatment of all stages of Lyme borreliosis. Parenteral therapy is usually only required for neuroborreliosis and cardiac disease with 3rd degree atrioventricular block, while oral antibiotics are sufficient for most other manifestations. In the past years a trend of prolongation of treatment can be noted. However, no evidence from controlled clinical studies is available to suggest that extension of treatment is beneficial. The risk of developing Lyme borreliosis in Switzerland is low. Prophylactic antibiotic treatment after a tick bite is not recommended. PMID- 16350539 TI - [Tick borne diseases]. AB - It is known for many years that tick-borne diseases have worldwide a high economical impact on farming industry and veterinary medicine. But only in the last twenty years the importance of such diseases were notified in human medicine by the medical community and the public with emerging of the tick borne encephalitis virus and the description of Borrelia burgdorferi. It is often forgotten that many other infectious agents as bacteria, virus, Rickettsia or protozoa can be transmitted by ticks. Such diseases are rarely diagnosed in Europe either they are overlooked and misdiagnosed or they are connected with special professional activities. The development of new regions for tourism with different out door activities (adventure trips, trekking, hunting) leads to an exposure to different tick borne diseases, which are often misdiagnosed. PMID- 16350540 TI - [Ectoparasitic diseases]. AB - Skin disorders are among the three most frequent health problems among returning travellers. Although ectoparasitic diseases are only a fraction of all skin problems, suspected lesions are often a reason to consult the GP after returning from low economy countries. Scabies presents in a variety of clinically different lesions and is a challenge to the physician, as are the residues of arthropode bites. A focused history and a careful clinical investigation is mandatory. Allergic reactions mirror the variety of individual modulation of immunological processes of ectoparasitic disorders. Further tropical and subtropical parasites such as tungiasis and myiasis are described in which simple measures result in dramatic improvement of major subjective suffering. PMID- 16350541 TI - [Worms and the lung]. AB - Due to international travel and migration helminthic infections are increasingly imported to countries were they are not endemic. A vast variety of helminthes may involve the lung. The lungs are either site of the infection or temporarily involved during maturation of helminthic larvae. In many affected patients the clinical picture is not specific and not typical. A thorough patient's history including a detailed travel history will lead to diagnosis, which must be confirmed by detecting the parasite either directly or with serological tests. The appropriate method depends on the parasite suspected and its life cycle within the human body. A combination of these methods will show the best results for most helminthes. As specific therapy is widely available for most helminthiases a precise diagnosis is important. PMID- 16350542 TI - [The worm in the brain]. AB - Cerebral signs of focalized infection or tumour with changing localisation or meningoencephalitis with a travel history should alert the clinician to consider the possibility of a worm infection. A detailed history of the stay abroad, of consumption of raw or with animal faeces contaminated food and of known preexisting helminthic disease gives further clues. The diagnostic approach compromises the check for eosinophilia in peripheral blood and CSF, serologic testing and CT scan or MRI examination. Anthelminthic treatment may induce a host reaction with inflammation and deterioration of the clinical picture and has to be performed cautiously and under protection with corticosteroids. PMID- 16350543 TI - [The worm in the liver]. AB - Patients with parasitic involvement of the liver are a small group of patients in European countries. In the differential diagnosis of "hepatic mass" "pain in the right upper abdomen" "biliary obstruction" or "abnormal liver function tests" parasites are, if at all, considered last. In most health care centers diagnostic and management experience is limited with respect to these diseases. They should therefore been taken care of by specialized centers. In most countries these are centers for tropical diseases. Important components of the work up of patients with parasitic diseases are: risk assessment (place of origin, travel and exposure), geographic distribution of the parasites, incubation period, prepatent period, laboratory investigations (eosinophilia, IgE, direct parasitological diagnosis) and imaging studies. Important parasites with liver involvement will be presented: schistosomiasis, echinococcosis, fascioliasis, toxocariasis. PMID- 16350544 TI - Contribution of occupational medicine to medical science. PMID- 16350545 TI - Chronic beryllium disease: a model for pulmonary sarcoidosis? AB - Chronic beryllium disease (CBD) and pulmonary sarcoidosis are two distinct chronic disorders, sharing the pathological lung hallmark of the non-caseating granuloma and the immunological feature of T cell activation at the site of disease. However, while CBD is a rare occupational disease in which the cause, i.e. the inhalation of beryllium, is well known since a long time, the etiology of sarcoidosis, which is far more common in the general population, is still unknown. Since granuloma formation requires the presence of an immunogenic initiating antigen, it has been hypothesized that sarcoidosis is an antigen triggered (auto)immune disease. Furthermore, while the study of large populations exposed to beryllium did made possible the identification of distinct genetic susceptibility factors in CBD, only recently the role of some genetic polymorphisms in sarcoidosis has been unraveled. Therefore, it seems likely that the advancement in the understanding of the immuno-pathogenesis of CBD will also help to design focused genetic studies to finally identify the etiology of sarcoidosis. Moreover, it is also possible that some cases of sarcoidosis are instead been caused by the inhalation of beryllium in genetically susceptible individuals. PMID- 16350546 TI - Asthma induced by isocyanates: a model of IgE-independent asthma. AB - Developments in the understanding of causes and natural history of asthma induced by isocyanates may allow improved preventive strategies for occupational asthma (OA), and may also lead to improved understanding of mechanisms involved in IgE independent nonoccupational asthma. Studies of genetic markers in OA induced by isocyanates suggest that HLA class II genes, glutathione S-transferase and NAT1 genotypes may predispose to development of this type of OA. Specific IgE antibodies against isocyanates are not always found in subjects with OA caused by isocyanates, leading most researchers to consider this type of OA, as a model of IgE-independent asthma. Evidence for cell-mediated immunity in OA induced by isocyanates has been provided by bronchoalveolar lavage, bronchial biopsy and induced sputum studies. The pathology of this type of asthma is similar to that of nonoccupational asthma, with cells such as eosinophils and T lymphocytes that exhibit signs of activation, and with thickening of the reticular layer of the basement membrane. Animal studies have shown that isocyanate asthma is driven primarily by CD4+ T cells and is dependent upon the expression of Th2 cytokines. However, animal models are not always reflective of human responses. OA induced by isocyanates similarly to nonoccupational asthma, is a multifactorial condition, and it is likely that complex gene-environment interactions play a role. Better understanding of these interactions is important for affected workers, and also has potential relevance for nonoccupational asthma. PMID- 16350547 TI - Silicosis and lung cancer: a fifty-year perspective. AB - The development of our studies on silica carcinogenesis and its mechanisms is reviewed. Starting from an analysis of the cellular reactions to silica in the pathogenesis of silicosis in the rat, followed by an analysis of the carcinogenic response to silica in the lungs of rats (but not in mice and hamsters), we went on to develop cellular models for culture and neoplastic transformation of rat alveolar epithelial cells. We studied the binding of silica to DNA, the generation of reactive oxygen species and the DNA damage mediated by hydroxyl radicals, showing marked differences among silica samples of varying purity. Then we investigated the role of peptides induced by silica in various cells, including cytokines and growth factors. Tumor necrosis factor (TNF)-alpha, which can cause activation of DNA transcription and is required for silica-induced fibrosis, was found to inhibit neoplastic transformation by quartz in cell cultures. Transforming growth factor (TGF)-beta was found to be produced in hyperplastic alveolar type II cells and to reach fibroblasts, macrophages and the connective tissue matrix adjacent to silicotic granulomas. Neuroendocrine cells and their peptides were found to be increased in alveolar and bronchiolar epithelia of silica lesions in rats, in contrast with mice and hamsters. Expression of adhesion molecules was found to be altered in silica-induced carcinogenesis and epithelial-mesenchymal transition was revealed by mesenchymal markers in the induced carcinomas. Promoter hypermethylation of adhesion genes in the induced carcinomas indicated a role for epigenetic mechanisms. PMID- 16350548 TI - Silica and the immune system. AB - This article collects the evidence that shows that the biological reactions to Silica are due to the stimulation of the Immune System. Both Innate and Adaptive Immunity are involved. The following sets of events take place sequentially: (1) Silica is recognized as a PAMP (pathogen-associated molecular pattern) by the Receptors of Innate Immunity; (2) This causes the stimulation first and then the death of the key cells of Innate Immunity (the macrophages); (3) While stimulated, macrophages produce cytokines (IL-1 and TNF) that stimulate fibroblasts; (4) The same and possibly other cytokines produced by silica- activated macrophages induce the maturation of dendritic cells, which are the connecting elements between the Innate and the Adaptive (lymphoid) Immune Systems; (5) It follows a polyclonal activation of the Adaptive Immunity; (6) The end result is the formation of fibro-hyaline tissue. In view of the double involvement of the Innate and the Adaptive Immune Systems and their cooperation in the stimulation of fibrosis, Silicosis can be considered as a "Collagen" Disease, related to other diseases of that group like Rheumatoid Arthritis, Lupus erythematosus and Scleroderma. Not surprisingly the incidence of these Diseases has been shown to be significantly increased in human exposed to Silica. PMID- 16350549 TI - Occupational allergic diseases as a clinical model to approach specific environmental reactivity. AB - Hyperreactivity to environmental factors is objectively expressed as respiratory, cutaneous and gastrointestinal disease. Approaching the diagnosis of an occupational disorder, a practical distinction was made between toxicity and allergy. The study of occupational allergic disease included particular procedures that led to standardized models and concepts. The contribution to the improvement of medical knowledge is reviewed according to selected experiences. The diagnostic aspects of asthma, rhinitis, dermatitis and urticaria are considered as regards to methodology of assessment of the occupational etiology with attention to demonstrative examples, which are worthwhile for the general medicine, too. Basic steps of risk agent identification, exposure assessment, threshold dose response measurement, allergen challenges and interaction are the original contribution of the occupational medicine to the diagnosis of allergic disorder. The clinical picture of asthma was clarified by the results of the specific bronchial provocation test, proving the important role of di-isocyanates and metal salts. Occupational rhinitis showed to be connected to asthma with predictive aspects in bakers' disease. Occupational dermatitis was linked to the development of experimental patch tests. Occupational urticaria included the concept of airborne contact allergy or nickel interactive food and occupational sensitivity. Occupational allergic diseases are emerging as a consequence of low environmental exposure, but they were remarkably studied in the past either for pathogenesis or for diagnostic procedures. Methods and acquisition are available also for the general medicine when an individual's specific reactivity is under investigation. PMID- 16350550 TI - Contribution of studies on renal effects of heavy metals and selected organic compounds to our understanding of the progression of chronic nephropathies towards renal failure. AB - Risk assessment for a number of workplace or environmental chemicals, especially heavy metals and industrial organic compounds, relies mostly on clinical and epidemiologic findings. The low incidence of chronic nephropathies raises methodological issues in carrying out and interpreting human data on the progression of early changes towards end-stage renal disease. To overcome such limitations of epidemiological studies, two main approaches have been explored: (i) human studies relying on biomarkers and (ii) experimental animal models. Animal experiments have been useful to characterize early changes, such as hyperfiltration, eventually leading to chronic renal failure. Animal studies provided insights into the mechanisms underlying microalbuminuria and microproteinuria. Such biomarkers of early changes, developed for use at the workplace, have then been used to monitor such chronic disorders and multifactorial diseases as diabetes and arterial hypertension. Another area where occupational medicine has provided evidence is the effectiveness of primary prevention over other possible approaches. Avoidance of exposure to heavy metals and volatile hydrocarbons and their derivatives, mainly in individuals with diagnosed renal disorders, remains the best approach towards a substantial reduction in the burden of renal diseases. PMID- 16350551 TI - Combined therapy with luteinizing hormone releasing hormone agonist (LHRHa) and growth hormone (GH) in central precocious puberty. AB - Luteinizing Hormone Releasing Hormone analogues (LHRHa) are considered the treatment of choice in central precocious puberty (CPP). Final height after therapy is usually higher than the predicted one before treatment, but about 60% of patients do not reach their genetic target. The association of Growth Hormone (GH) with LHRHa could be useful in a subgroup of children with CPP and low growth velocity in order to obtain a higher final height with a complete expression of genetic potential. Moreover, in children with early puberty adopted from developing countries, predicted adult height is significantly greater when the combined therapy, GH and LHRHa, is employed. In this paper the rationale for adding GH to LHRHa is discussed and the clinical reports from the literature are presented. PMID- 16350552 TI - Breakfast habits of 1,202 northern Italian children admitted to a summer sport school. Breakfast skipping is associated with overweight and obesity. AB - Very little is known about the differences in breakfast of children performing physical activity in the morning. This paper analyzed the breakfast habits of 747 boys and 455 girls, distributed in 2 homogeneous age groups, 6-10 and 11-14 year old, participating in a Summer Sport School. Children were asked whether, when, where, how and with whom they consumed breakfast; who prepared meals; what they ate and drank; what they did during breakfast. Weight, height and BMI were recorded. Seventy-eight percent of children usually had breakfast, but 22% reported skipping breakfast. In the non-breakfast consumer subjects, 27.5% were overweight and 9.6% obese vs 9.1 and 4.5% respectively in breakfast eaters. Bakery products (76%) and milk (71%) were the most frequently consumed foods. Only 15% of parents encouraged their children to consume additional foods at breakfast before exercising, and 42% of children believed that this extra nutrition was unnecessary. In 80% of cases, the chief decision-maker for breakfast was the mother, the father played little part (1%). During breakfast, 48% of children ate and drank in silence, 26% played with brothers and sisters, 18% watched television and only 8% talked with parents. A high prevalence of over weight and obesity was found among non-breakfast consumer children. Breakfast omission in children exercising conflicts with their increased energy requirements and may be connected to the trend of parents and children to under estimate the importance of breakfast for nutritional balance and for environment promotion of physical performance. PMID- 16350553 TI - Intravenous contrast material administration in multislice computed tomography coronary angiography. AB - RATIONALE AND OBJECTIVES: to compare contrast material (CM) administration protocols in non-invasive coronary angiography (CA) using a 16-row multislice CT (16-MSCT). METHODS: 45 patients undergoing CA with 16-MSCT were divided into three CM protocols: group 1 (140 ml@4ml/s), group 2 (140 ml = 60 ml@5ml/s + 80 ml@3ml/s), and group 3 (100 ml@4ml/s). The attenuation at the origin of the coronary vessels was assessed. Three regions of interest were evaluated: 1) ascending aorta (ROI1); 2) descending aorta (ROI2); 3) pulmonary artery (ROI3). The resulting time-density curves generated the average attenuation and the slope of bolus geometry. RESULTS: the attenuation at the origin of the coronary vessels, and the average attenuation of bolus geometry were not significantly different (p > 0.05). The slope of bolus geometry was in ROI1 and ROI2 significantly lower for group 2, in ROI3 significantly lower for group 3 (p < 0.05). CONCLUSION: 100 ml of CM provide the same attenuation in 16-MSCT CA as mono- or multi-phasic 140 ml protocols. PMID- 16350554 TI - Endoscopic treatment of esophageal and colo-rectal fistulas with fibrin glue. AB - So far, the use of fibrin glue has been limited to the treatment of anal, recto vaginal and enterocutaneous fistulae. Between 1991 and 2003 we performed the treatment of anastomotic leaks of the upper and lower gastro-intestinal tract with fibrin glue in 13 selected patients. In our experience the treatment with fibrin glue has been proved to be effective in the selected cases. If the fistulae clinically occured 7 days after surgery a higher number of endoscopic sessions were necessary than in patients with earlier appearance of anastomotic leakage. The utilization of fibrin glue for the endoscopic management of anastomotic leakages after surgery can be successful and safe if applied in selected patients. PMID- 16350555 TI - Acute coronary syndromes without persistent st-segment elevation: advances in surgical revascularization. AB - Coronary artery bypass grafting (CABG) still plays a fundamental role in the management of acute coronary syndromes. The aim of this study is to report the experience of our center in the treatment of patients with acute coronary sindromes without persistent ST elevation urgently operated on with CABG, and to discuss surgical problems related. Two-hundred and six patients were urgently operated on for CABG for acute coronary syndromes without persistent ST-segment elevation from January 2001 to February 2003. The majority of them had three vessel coronary disease (72%) and left main stem disease occurred in 20% of the patients. Mean LVEF (left ventricular ejection fraction) was 54 +/- 12% whereas 9% of the patients had a LVEF < 40%. Twenty-one patients (10%) received glycoprotein IIb/IIIa receptor inhibitors and 35 (17%) received intravenous heparin therapy before surgery. Mean interval time between the onset of symptoms and surgery was 16 +/- 10 days (range 4-50). In-hospital mortality was 2% (4 patients). Perioperative AMI (acute myocardial infarction) occurred in 4% (8 patients) and a transient low cardiac output syndrome in 27 patients (13%). Bleeding requiring surgery occurred in 1% of the patients. Transient respiratory insufficiency was present in 12 patients (6%) and acute renal failure in 8 patients (4%). Mean I.C.U. time was 2.4 days (1-17). Urgent CABG for acute coronary syndromes shows a low risk for in-hospital mortality and morbidity. In acute patients arterial grafts are not detrimental for the outcome, but are recommended in the absence of contraindications to improve long-term results. In spite of recent developments in cardioplegic cardiac arrest, optimal myocardial protection against perioperative myocardial infarction still remains a challenge. PMID- 16350556 TI - Combined arthroscopic and radioscopic management of tibial plateau fractures: report of 18 clinical cases. AB - Tibial plateau fractures are complex lesions capable of causing severe consequences if not appropriately treated. They are often the result of a high energy trauma and, not rarely, are associated with significant soft-tissue and intra-articular injuries. Different therapeutic options can be managed in the treatment of these lesions. Minimally invasive surgery offers several advantages compared to other surgical techniques and allows, with less additional soft tissue damages, good reduction and stable fixation of the fracture. In this study we assessed the results of the combined arthroscopic and radioscopic assisted reduction and internal fixation of tibial plateau fractures in 18 patients affected by Schatzker type I, II, III, IV fractures. According to Hohl's and Rasmussen's grading system, 16 out of 18 patients scored a satisfactory result. We experienced no complications due to arthroscopy. PMID- 16350557 TI - Ureteral hemangioma: a clinical case report. AB - The most greater part of the tumors ureteralis are of malignant nature, they are verified especially in the elderly patient and they have a preference for the distal third of the ureter. Of the tumors benign ureteral, the emangioma is a very rare vascular neoplasm. The symptoms aspecificis of presentation are: hematuria, colic type pains, hydronephrosis and bladder inflammation. Thanks to the refinement of the radiological techniques (Urography, CTscan and eventually the ascending pyelography) and also to the simultaneous aid of the anatomopathologist, it is possible to obtain an accurate description of this pathology in a precocious stage in order to carry out a conservative approach. The peculiarity of the clinical case from us described depends on the fact that it deals with an occasional find, reaches our observation for appearance of severe hydronephrosis caused by ureteral stones, resolved him with methodic endoscopic. PMID- 16350558 TI - Two clinical cases of insulinoma misdiagnosed as psychiatric conditions. AB - We describe the clinical cases of two male subjects, respectively 73 and 41 years old, affected with insulinoma, misdiagnosed for many years as psychiatric subjects and scarcely responsive to therapy with benzodiazepins, antidepressants and antiepileptics. Although Whipple's triad was satisfied in both subjects since the onset of symptoms, they had to wait a long time for the correct diagnosis and appropriate treatment. Once the surgical therapy was carried out, our subjects recovered totally and ended the psycho-drugs. PMID- 16350559 TI - Operating risks evaluation in posterior transolecranon surgical approach: the traffic lights model. PMID- 16350560 TI - Mineral and water content of the fat-free body: effects of gender, maturation, level of fatness, and age. AB - Variability associated with the effects of gender, level of fatness (LOF), maturation (ML), and age on the mineral (MFFB) and water (WFFB) content of the fat-free body (FFB) was investigated in 188 males and 144 females, aged 8 to 17 years. Measures of body density, bone mineral content, total body water, and skin fold thicknesses were obtained. FFB was calculated using a multicomponent model based on body density and adjusting for variability in body water and mineral content. Subjects were classified by ML as prepubescent, pubescent, and postpubescent. Subjects were also grouped by LOF as lean, average, and obese based on percentile rankings from the age-related norms of the National Children and Youth Fitness Study. Least squares multiple regression analysis using weighted orthogonal contrasts to account for sample size differences revealed significant (p<.01) gender, ML, gender x LOF, and gender x ML effects for MFFB; however, no significant (p>.05) effects between, or within, gender were noted for WFFB. Significant ML, and nearly significant (p>.056) LOF x ML, effects were found for MFFB in the males. In contrast, only ML was significant within the female sample for MFFB. These results were upheld when age, rather than ML, was used in the analysis. The overall relative increase in MFFB across ML was greatest in the obese (17.4%, 9.3%) and least in the lean (11.3%, 6.3%) males and females, respectively. A significant increase in MFFB across ML was observed in both genders; however, the pattern and magnitude of increase is dependent upon LOF considered. Hence, consideration of gender, ML, LOF and age is essential in estimating body composition in children. PMID- 16350561 TI - Gallstone formation and weight loss. AB - Obesity is associated with increased bile stasis and cholesterol saturation, and an increased risk of gallstone development. Conversely, bile composition is normalized following reduction in body weight. It would appear advantageous to promote weight loss in obesity, which would reduce the predisposition to gallstone formation. Despite the potential health benefits of weight reduction, very-low-calorie diets appear to increase the risk for cholesterol crystal and gallstone formation. The incidence of gallstone formation seems to be dependent on the degree of caloric restriction, the rate of weight loss, and the duration of the dietary intervention. Thus, faster rates of weight loss for longer periods of time are associated with increased risk. Available data obtained from prospective studies of subjects during active weight loss suggest that newly formed gallstones occur within 4 weeks and with incidence rates 15 to 25-fold higher than in the general obese population. The stones produce symptoms in approximately one-third of the subjects, of whom approximately one-half will undergo surgery. Proposed mechanisms underlying gallstone formation during weight reduction include bile stasis due to reduced caloric intake, increased biliary cholesterol saturation secondary to increased cholesterol mobilization, and increased nucleation due to changes in bile arachidonate and glycoprotein concentrations. Data are lacking on the effects of gradual rates of weight loss and risk of gallstone formation. PMID- 16350562 TI - The Gulstonian Lectures. 1850. PMID- 16350563 TI - Commentary on paper by Chambers. PMID- 16350564 TI - Two preadipocyte subtypes cloned from human omental fat. AB - To determine whether the characteristics of preadipocytes derived from human fat are uniform or variable, we developed methods for culturing and differentiating cloned human preadipocytes. Individual human omental preadipocytes were cultured for six weeks. The number of cells varied considerably among clones derived from the same subject, implying that human preadipocytes vary in replicative capacity. Indeed, two cell subtypes were found in human omental fat; one type replicated slowly and the other was capable of extensive replication. Cells of both subtypes were capable of differentiation into adipocytes, confirming that both subtypes were preadipocytes. When rat perirenal and epididymal preadipocytes were cloned, a slowly replicating and an extensively replicating preadipocyte subtype were also found. It is proposed that preadipocytes of the rapidly and the slowly replicating subtypes may be at different stages along the pathway between uncommitted precursor cells and differentiated adipocytes. PMID- 16350565 TI - Fluoxetine: a randomized clinical trial in the maintenance of weight loss. AB - Because current weight-reduction treatments have considerable recidivism, a therapy that could help patients maintain weight loss would be of benefit. A six center, randomized, double-blind trial compared the effects of the specific serotonin uptake inhibitor, fluoxetine hydrochloride, and placebo on maintenance of weight loss. Obese outpatients who had lost > or = 3.6 kg after 8 weeks of single-blind fluoxetine 60 mg/day in the qualification phase (N=317 [70.4% of patients entered]; mean +/- standard deviation [SD] weight loss, 6.8 +/- 2.8 kg) were randomly assigned to fluoxetine 20 mg/day (N=104), fluoxetine 60 mg/day (N=106), or placebo (N=107) for 40 weeks (maintenance phase). Patients received minimal nutrition/dietary counseling. Qualification phase clinic visits were biweekly; maintenance phase visits were monthly for 4 months, then bimonthly for 6 months. Patients treated with fluoxetine 60 mg/day continued to lose weight for 8 additional weeks (16 weeks total; maximum mean +/- SD weight loss, 7.2 +/- 4.6 kg); those treated with fluoxetine 20 mg/day or placebo began to regain weight. Mean weights remained below baseline values at week 48 (all groups); treatment differences were not statistically significant. Study completion rates were comparable (fluoxetine 20 mg/day, 67.3%; fluoxetine 60 mg/day, 56.6%; placebo, 67.3%; p = 0.175). Among commonly reported adverse events (> 10% incidence), only asthenia was reported statistically significantly (p < 0.050) more frequently with fluoxetine than with placebo. Few patients discontinued for any single adverse event. Fluoxetine 60 mg/day was effective for a longer period than fluoxetine 20 mg/day or placebo in maintaining weight loss. Overall, fluoxetine was safe and well tolerated. PMID- 16350566 TI - Effects of dexamethasone on primarily cultured newly differentiated rat adipocytes from different adipose tissue regions. AB - The effects of dexamethasone (dex) on newly differentiated adipocytes in primary culture derived from mesenteric, retroperitoneal, epididymal, and inguinal subcutaneous adipose tissues of male rats were studied. The degree of differentiation was similar in these adipose precursor cells derived from different regions as assessed by lipoprotein lipase (LPL) activity, an early marker of adipocyte differentiation. LPL activity was increased by addition of dex, and no differences in degree of activation were observed in cells from different adipose tissue regions. Development of both basal and isoproterenol stimulated lipolysis was also similar in adipose precursor cells from different adipose tissue regions. Dex addition enhanced the isoproterenol-stimulated lipolysis with no regional differences. Studies of binding of [3H]-dex showed no regional differences in either binding affinity or maximal binding capacity. It is concluded that dex stimulates both LPL activity and lipolytic activity in newly differentiated rat adipocytes in primary culture. This seems, however, not to vary in magnitude in cells obtained from different adipose tissue regions. This might be due to the apparent similarity of number and affinity of glucocorticoid binding sites. Regional variations in glucocorticoid regulated LPL and lipolytic activity in adipose tissue might therefore not be due to inherent differences between adipocytes. PMID- 16350567 TI - Altered brown adipose tissue and Na,K pump activities during diet-induced obesity and weight loss in rats. AB - Brown adipose tissue (BAT) thermogenesis is an uncoupled ATPase-independent thermogenic mechanism. Ion transport by the Na,K pump is an ATPase-dependent thermogenic mechanism. Both have been proposed as mechanisms of altered energy expenditure during states of dietary energy surfeit and deficit. Our aim was to study these mechanisms during diet-induced obesity and weight loss. Over 36 weeks rats were fed lard- or tallow-based diets (63% energy as fat), or a control diet (12% energy as fat). During periods of restriction rats were fed 50% of the energy intake of controls in the form of a control diet. Several components of thermogenic response increased in rats eating high fat diets and decreased following dietary restriction. BAT activation occurred, particularly with a lard based diet, as indicated by increased GDP binding and uncoupling protein (UCP) content. Na,K pump activity in thymocytes increased with the feeding of both high fat diets at some time points. Plasma T3 level increased in rats eating the lard based diet and decreased with dietary restriction regardless of previous diet. Resting metabolic rate (RMR) of the animals was unchanged despite increases in these thermogenic components and was decreased in all groups following dietary restriction. Our results indicate a lack of any major role for activated BAT thermogenesis in mitigating the extent of the obesity induced by the high fat diets. The reasons for the differences in response to the two different sources of saturated fat, lard, and tallow, are not clear. PMID- 16350568 TI - Level of preadipocyte growth factor in rat adipose tissue which specifically permits the proliferation of preadipocytes is affected by restricted energy intake. AB - We previously reported the presence of a protein growth factor in rat adipose tissue which specifically permits the proliferation of 3T3-L1 and Ob1771 preadipocytes [Biochem. Biophys. Res. Commun. 1990;171:905-912, ref. 1] and which is hereinafter referred to as PAGF (preadipocyte growth factor). In this study, the effects of long-term restricted energy intake on the PAGF activity in rat epididymal and perirenal adipose tissue toward 3T3-L1 preadipocytes were investigated. When rats were subjected to restricted energy intake for three weeks, PAGF activity increased with energy intake. The body weight, epididymal and perirenal fat depot weights and glycerol 3-phosphate dehydrogenase activity also increased with the energy intake, whereas the lactate dehydrogenase activity remained almost constant in all energy intake groups. These results suggest that the PAGF in fat depots functions in response to energy intake and contributes to the de novo formation of adipocytes and the growth of adipose tissue. This factor may provide a useful tool for further elucidation of the relationship between energy storage in adipose tissue and adipose tissue development. PMID- 16350569 TI - Exercise and obesity. AB - This paper reviews succinctly the evidence for a role of regular exercise in the prevention and the treatment of obesity and of its metabolic complications. Seventeen propositions relevant to an understanding of the topic are considered. The evidence suggests that regular exercise can be an important factor in the development of sustained negative energy balance conditions provided the volume of activity is high. This implies a program of low to moderate intensity exercise performed on an almost daily basis for at least one hour per session. To induce significant weight and fat losses and to treat overweight and obese patients, compliance to the program for several years becomes a necessity. Exercise increases lipid substrate oxidation and may favor carbohydrate intake for the same amount of energy intake. The acute effects of exercise on resting metabolic rate are well documented, but the long-term influences of exercise training seem to be small and are rapidly suppressed with the cessation of training. The obese benefits also from a regular exercise regimen in terms of improved insulin sensitivity, lipid and lipoprotein profile, and blood pressure, as well as reduced risk of death. Regular exercise, such as walking, is a healthy course of action for the overweight or the obese patients. PMID- 16350570 TI - Commentary on Banting letters. PMID- 16350571 TI - Letter on corpulence, addressed to the public. 1869. PMID- 16350572 TI - Plasma cortisol levels increase with age in obese subjects. AB - In order to assess the influence of age, sex, and body mass on plasma cortisol concentrations, we measured the 24-hour Integrated Concentration (IC) of cortisol (F) in 36 obese subjects (16 males, 20 females) aged 5.3-56.4 years, BMI = 35.5 +/- 7.3 kg/m2 and compared with 119 nonobese subjects, body mass indices (BMI) 21.2 +/- 2.7 kg/m2, aged 8.8-66.2 years (55 males, 64 females). Subjects were nondiabetic, normotensive, without history of psychiatric illness, and otherwise in good health. IC studies were performed using a continuous blood withdrawal methodology, and IC-F was assayed from the 24 hour pooled sample by a protein binding method. The effect of age and gender on IC-F was analyzed by multivariate regression. In the nonobese group there was no effect of age or sex on IC cortisol levels, the mean IC-F = 173.8 +/- 44.1 nmol/L. A statistically significant but weak negative effect of BMI on IC-cortisol (r=-.18, p<0.05) was present. In the obese subjects there was a significant increase in IC-cortisol levels with age IC-F(nmol/L) = 2.76 x age(years) + 85.0 (r2=.36, p<0.0001). IC cortisol levels tended to be lower in obese males than females when controlled for age (p<0.05). We conclude that in nonobese subjects IC-F levels are independent of age and gender. However, there is a significant increase of IC cortisol levels with age in obese individuals. The observed increase of IC cortisol with age may contribute to metabolic complications of obesity. PMID- 16350573 TI - Fat distribution and body weight. PMID- 16350574 TI - Visceral obesity: a "civilization syndrome". AB - The controversial question of the relationship between obesity and disease has been considerably clearer after the demonstration in several prospective, epidemiological studies that the subgroup of central, visceral obesity is particularly prone to develop cardiovascular disease, stroke, and non-insulin dependent diabetes mellitus. Visceral obesity is associated with multiple central endocrine aberrations. The hypothalamo-adrenal axis is apparently sensitive to stimuli, sex steroid hormone secretion blunted, and hyperandrogenicity is found in women. In addition, there seem to be signs of central dysfunctions in the regulation of hemodynamic factors after stress, and growth hormone secretion appears to be particularly blunted. Several of these endocrine abnormalities are associated with insulin resistance, particularly glycogen synthesis in muscle. Fiber composition with low type I/type II ratio might be secondary to the prevailing hyperinsulinemia, but low capillary density in muscle may well be of importance. In combination with elevated turn-over of free fatty acids (FFA) this will probably provide powerful mechanisms whereby insulin resistance is created. Portal FFA, from the highly lipolytic visceral depots may, in addition, affect hepatic metabolism to induce increased gluconeogenesis, production of very low density lipoproteins as well as to perhaps inhibit clearance of insulin. By these mechanisms a Metabolic Syndrome Visceral adipocytes seem to have a high density of several steroid hormone receptors, directing steroid hormone effects particularly to these depots. The net effect of cortisol is apparently a stimulation of lipid storage, with opposing effects of sex steroid hormones which also facilitate lipid mobilization, regulations most often found at the gene transcription level. Growth hormone inhibits cortisol effects on lipid accumulation, and amplifies the lipid mobilizing effects of steroid hormones. The combined perturbations of hormonal secretions will therefore probably direct triglycerides toward visceral depots. Circulatory and nervous regulatory mechanisms require, however, more attention. The multiple central endocrine and nervous aberrations of visceral obesity suggest neuroendocrine dysregulations, and have features characteristic of the hypothalamic arousal seen after certain types of stress, alcohol intake, and smoking. Such factors can be traced to subjects with visceral fat accumulation. Standardized stress, eliciting a "defeat reaction" in primates is followed by an apparently identical syndrome. This integrated picture of the multiple symptoms of visceral obesity is based on epidemiological, clinical, experimental, cellular, and molecular evidence. The ingredients of positive energy balance, including physical inactivity, stress, smoking, and alcohol consumption are frequent features of modern, urbanized society. Visceral obesity may therefore be an expression of a "Civilization Syndrome." PMID- 16350575 TI - Commentary on Atwater classic. PMID- 16350576 TI - An experimental inquiry regarding the nutritive value of alcohol. 1902. PMID- 16350577 TI - Androgen treatment of abdominally obese men. AB - Middle-aged men with abdominal obesity were treated in a double-blind study with moderate doses of transdermal preparations of testosterone (T), dihydrotestosterone (DHT), or placebo. This resulted in moderately elevated T concentrations and marked decreases in follicle stimulating and luteinizing hormones in the group treated with T, while the DHT group showed elevated DHT, markedly lower T values, and less diminution of gonadotropin concentrations. In the group treated with T visceral fat mass decreased (measured by computerized tomography) without significant changes in other depot fat regions. Lean body mass did not change. In the group treated with T, glucose disposal rate, measured with the euglycemic hyperinsulinemic clamp method, was markedly augmented. Plasma triglycerides, cholesterol, and fasting blood glucose concentrations as well as diastolic blood pressure decreased. There were no such changes in the DHT or placebo treatment groups. The men treated with T reported increased well-being and energy. In none of the groups did prostate volume, specific prostate antigen concentration, genito-urinary history, or urinary flow measurement change. It is suggested that supplementation of abdominal obese men with moderate doses of T might have several beneficial effects. PMID- 16350578 TI - Duration of improved muscle glucose uptake after acute exercise in obese Zucker rats. AB - Skeletal muscle is insulin resistant in the obese Zucker rat. Endurance training reduces muscle insulin resistance, but the effects of a single acute exercise session on muscle insulin resistance in the obese Zucker rat are unknown. Therefore, insulin responsiveness of muscle glucose uptake was measured in 15 week-old obese rats either 1, 48, or 72 hours after two hours of intermittent exercise (30:30 min; work:rest). Hindlimbs of sedentary lean (LS) and obese (OS) rats and exercised obese (OE) rats were perfused after a 10-hour fast under both basal (0 mU x ml(-1)) and maximal (20 mU x ml(-1)) insulin concentrations to measure net glucose uptake. Insulin responsiveness of net glucose uptake was significantly reduced in OS compared to LS (8.5 +/- 1.6 vs 15.3 +/- 2.0 micromol x g(-1) x h(-1), respectively). Compared to OS, insulin responsiveness of net glucose uptake was significantly increased by 56% and 80% at 1 hour and 48 hours after acute exercise. However, 72 hours after acute exercise, the increased insulin responsiveness of net glucose uptake was no longer evident. These results indicate that improved responsiveness of muscle glucose uptake persists for at least 48 hours after two hours of acute intermittent exercise in 15-week-old obese Zucker rats. PMID- 16350579 TI - Androgens and body fat distribution in men. PMID- 16350580 TI - Binge eating disorder: current knowledge and future directions. AB - Binge eating disorder (BED) is a newly characterized eating disorder that encompasses individuals who have severe distress and dysfunction due to binge eating, but who do not regularly engage in inappropriate compensatory behaviors. While relatively uncommon in the general community, BED becomes more prevalent with increasing severity of obesity. BED is associated with early onset of obesity, frequent weight cycling, body shape disparagement, and psychiatric disorders. These associations occur independent of the degree of obesity. Although many individuals with BED have good short-term weight loss regardless of treatment modality, as a group they may be prone to greater attrition during weight-loss treatment and more rapid regain of lost weight. Current treatments geared toward binge eating behaviors include antidepressant medications, cognitive behavioral psychotherapy, and interpersonal psychotherapy; however, these treatments have little efficacy in promoting weight loss, and only modest success in long-term reduction of binge eating. As a significant proportion of obese individuals entering weight-loss treatment and research programs are likely to meet criteria for BED, those conducting clinical research should be aware of this distinct subgroup and determine the contribution of BED to outcome measures. PMID- 16350581 TI - Commentary on classics of obesity. 4. Hypothalamic obesity. PMID- 16350582 TI - The Frohlich syndrome: report of the original case. 1939. PMID- 16350583 TI - Tumor of the body of the pituitary without acromegaly and with arrest of development of the genital organs. 1900. PMID- 16350584 TI - Neuropathology communication from Dr. Mohr, privat docent in Wurzburg. 1840. PMID- 16350585 TI - The influence of human growth hormone (GH) and thyroxine (T4) on the differentiation of adipose tissue in the fetus. AB - Late term fetuses from genetically obese dams have slightly larger fat cells, greater adipose tissue lipoprotein lipase (LPL) activities, elevated levels of thyroid hormones, and depressed growth hormone (GH) levels when compared to fetuses from lean dams. We have investigated the influence of thyroid hormone and GH status per se on these and other adipose tissue traits by chronically treating hypophysectomized (hypox) fetuses (day 70) between day 90 and 105 of gestation with either thyroxine (T4) or human GH. Treatment with T4 decreased body weights (P<.05), increased serum T4 levels (P<.05), and enhanced skin and hair development (P<.05). Quantitative analysis of sections of perirenal and subcutaneous adipose tissue indicated that T4 increased LPL activity (P<.05), slightly increased fat cell size, and more than doubled (P<.05) lipid accretion. A hypox induced deficit in fat cell cluster number in the outer layer of subcutaneous tissue was normalized by T4 (P<.05). Conversely, human GH (hGH) treatment had no influence on body weight, increased serum hGH levels, decreased fat cell size (P<.05) and LPL activity (P<.05) but had no influence on lipid accretion. Quantitative analysis of adipose tissue sections provided direct and indirect evidence of a "critical" or "sensitive" period between 90 and 105 days, since fetal hypox at day 70 severely impeded preadipocyte recruitment/replication during this period. Furthermore, T4 but not GH effectively normalized this hypox induced deficiency in preadipocyte development. Therefore, T4 may have a major role in preadipocyte recruitment/replication during late fetal life. PMID- 16350586 TI - Body mass and risk of hip fracture among a national cohort of postmenopausal white women: a reanalysis. AB - To assess the prospective relationship between body mass and rate of hip fracture, we analyzed data from 2,285 postmenopausal women, aged 50-77 years, who responded to the first National Health and Nutrition Examination Survey (NHANES I) in 1971-1974 and the 1982-84 interview of the National Health Epidemiologic Follow-up Study (NHEFS). We plan to add to the previous analysis of these same data by Farmer et al. by 1) including retrospective information on estrogen replacement therapy; 2) increasing the follow-up time by 5 years and using only hospital-verified cases of hip fracture from the 1987 Health Care Facilities Stay data tape; and 3) limiting the analysis to postmenopausal white women, ages 50-77 years. Results of the multivariable Cox regression modeling showed that women with a baseline body mass index in the highest quartile (> 37 kg/m1.5) experienced a 70% lower rate of hip fracture compared with women in the lowest quartile (< or = 28.7 kg/m1.5)(RR=0.32; 95%CI:0.12, 0.82). Age was positively related to the risk of hip fracture, with the rate over 20% higher per year of age (RR=1.21; 95%CI:1.13, 1.29). Although reported education level, smoking history, physical activity level, and estrogen replacement were significantly (p < 0.0001) associated with body mass index, these covariates were not related to hip fracture in the multivariable analysis. Our findings corroborate earlier results using this same data source, and suggest that interventions aimed at preserving lean mass and consequent bone integrity should be encouraged among women before menopause and maintained through older adulthood. PMID- 16350587 TI - Structural changes in the subcutaneous compartment in morbid obesity. AB - Most of the excess fat that accumulates in the morbidly obese is accommodated in the subcutaneous compartment (SCC), specifically in the adipose tissue lobules of the panniculus adiposus which enlarge and expand the SCC. However, the fibroelastic septa that are attached to the skin on one side of the SCC and to the deep fascia on the other invest each lobule and thereby offer resistance to the enlarging lobules. We report observations made during pathologic examinations of the abdominal wall SCC that were surgically excised by abdominal dermo panniculectomy from 48 morbidly obese subjects after gastroplasty and weight loss. Lobules were large but varied notably upwards in size and, to some extent, shape. Simultaneously upon incision of the SCC, the lobules bulged above and the septa retracted below the same cut surfaces. Light microscopy revealed disruptions of septal elastic fibers, calcification of septa and septal arteries and necrosis of adipocytes, all sparsely distributed. Certain questions and facts emerge from these observations. Are the variations in lobule sizes normal, or do they reflect uneven fat deposition or mobilization during weight gain or loss respectively? Is there any correlation between lobule and adipocyte sizes? The tightly coupled phenomena of lobule-bulging and septal-retraction indicate that the lobules and septa are subjected to compressional and tensile stresses respectively in the intact, non-incised SCC. The histologic changes might be consequences of these stresses. Studies of the variations in body fat distribution and in adipocyte size have contributed to an understanding of obesity pathogenesis. The relevance of the present findings is unknown at this time, but these appear to have potential implications for studies of the morphogenesis of obesity. Confirmation of these findings, particularly the lobule variations, is required so that these can be compared with those in never-obese subjects and in morbidly obese subjects without prior weight loss. PMID- 16350588 TI - Weight cycling: the public concern and the scientific data. PMID- 16350589 TI - Commentary on classics in obesity. 5. Fat cell theory and units of knowledge. PMID- 16350590 TI - Microscopial researches into the accordance in the structure and growth of animals and plants. 1847. PMID- 16350591 TI - Observations on the development of the fat vesicle. 1849. PMID- 16350592 TI - An international nursing community of scholars? a community of scholars for international nursing science? or both? PMID- 16350593 TI - The leadership challenge. PMID- 16350594 TI - Nurses' clinical engagement: a study from an acute-care setting in Norway. AB - This study investigates the nature of nurses' clinical engagements and courses of action in acute-care settings in Norway. A qualitative descriptive design with a convenience sample of 6 registered nurses was used. Data were collected via participant observation, in-depth interviews during 3 full shifts for each nurse, and nursing documents about the patients. Data analyses used the method suggested by Atkinson (1992) that coalesces analyses of parts with wholes. Findings revealed that nurses' clinical engagements are multidimensional and consist of 3 sorts of activities: (a) nurses' movements and encounters with patients, (b) the process of knowing patients, and (c) clinical actions addressing patients' specific needs/requests. These activities constituted most nursing work when the nurses were providing care for hospitalized patients. PMID- 16350595 TI - An ontological view of advanced practice nursing. AB - Identifying, developing, and incorporating nursing's unique ontological and epistemological perspective into advanced practice nursing practice places priority on delivering care based on research-derived knowledge. Without a clear distinction of our metatheoretical space, we risk blindly adopting the practice values of other disciplines, which may not necessarily reflect those of nursing. A lack of focus may lead current advanced practice nursing curricula and emerging doctorate of nursing practice programs to mirror the logical positivist paradigm and perspective of medicine. This article presents an ontological perspective for advanced practice nursing education, practice, and research. PMID- 16350596 TI - Shifting patterns of practice: nurse practitioners in a managed care environment. AB - The purpose of this qualitative study was to uncover patterns across nurse practitioner (NP) experiences that contribute to understanding their perceptions of managed care, how it affects daily practice, and how NPs respond to a changing managed care workplace. In-depth interviews were conducted with 14 NPs representing primary care, specialty, and independent practices. Over an 18-month period, data collection and analysis occurred simultaneously using standard methods of purposive sampling, constant comparison, memoing, and member checks. This study illuminates the tension NPs experience between a business and a professional ethic and the strategies they use to reconcile this difference with core nursing values. Type of setting, workplace dynamics, and length of time in practice contributed to variation in NP perspectives. PMID- 16350597 TI - The role of mentoring in developing African American nurse leaders. AB - The primary purpose of this study was to explore the role of mentoring in the development of African American nurses who have achieved leadership positions in baccalaureate and graduate nursing programs. This study also explored similar and dissimilar mentoring experiences in same-race versus cross-race mentoring relationships. The theoretical framework for this study was Levinson's adult developmental theory. A sequential mixed design was utilized. Forty-seven African American nurse leaders participated in Phase 1, and 10 of the 47 were interviewed in Phase 2. The findings showed that mentoring plays a role in the personal and professional development of African American nurse leaders in baccalaureate and graduate nursing programs. Moreover, the relevance of race varies in both same race and cross-race mentoring relationships. PMID- 16350598 TI - The social construction and conceptualization of sexual health among Mexican American women. AB - Research on the social construction and contextualization of sexual health among Latina women is sparse. This ethnographic study was aimed to bridge this gap by exploring sexual health experiences and behaviors of a Latina subgroup, Mexican American women. A series of individual semistructured, in-depth interviews were conducted with 20 Mexican American women from a sexual and reproductive health clinic in a large Southwestern city with a predominantly Mexican American population. Through thematic analysis, sacrificing of self, sexual silence, and taking control of self emerged as themes. These themes reflect the context within which construction of sexual health occurs and sexual health behaviors are influenced. Study findings are discussed within a socio-cultural framework for sexual health promotion intervention for both adolescent and adult Mexican American women. PMID- 16350599 TI - Diagnosis and treatment of attention-deficit/hyperactivity disorder (ADHD) in African-American and Hispanic patients. PMID- 16350600 TI - Overcoming challenges in the diagnosis and treatment of attention deficit/hyperactivity disorder in African Americans. AB - The incidence of ADHD appears to be similar in African Americans and white populations. However, fewer African-American than white children are diagnosed and treated for ADHD. Reasons for this disparity have not been fully elucidated; causes are most likely complex. Whereas certain barriers to treatment are driven by patients and their families, others are due to limitations in the healthcare system. Patient-driven obstacles to care include inadequate knowledge regarding the symptoms, treatment and consequences of untreated ADHD and fear of overdiagnosis and misdiagnosis. A survey conducted to explore cultural differences between African-American and white respondents found that African Americans were more likely than whites to be unfamiliar with ADHD. In addition, African Americans felt that they were diagnosed with ADHD more often than whites and that teachers blamed ADHD for learning or behavior problems more often in African Americans. Health system barriers include a lack of culturally competent healthcare providers, stereotyping/biases and failure of the clinician to evaluate the child in multiple settings before diagnosis. Strategies to overcome these challenges include increased dissemination of ADHD information through community events; improved training of clinicians in cultural competence; and open communication among parents, clinicians and school personnel. PMID- 16350601 TI - Multicenter, randomized, open-label study of OROS methylphenidate versus atomoxetine: treatment outcomes in African-American children with ADHD. AB - The Formal Observation of Concerta versUs Strattera (FOCUS) study was conducted to assess, in children with ADHD, treatment outcomes with Concerta [OROS methylphenidate (MPH)], a once-daily controlled-release medication, and Strattera, (atomoxetine), a selective noradrenaline reuptake inhibitor, Because of the lack of data in minority groups treated for ADHD, the present subgroup analysis was conducted to determine the effectiveness and tolerability of ADHD treatments in African-American patients who were randomized to OROS MPH (n=125) or atomoxetine (n=58) during the FOCUS study. At the end of the study, the mean dose of OROS MPH was 32.8 +/- 10.9 mg and that of atomoxetine was 1.1 +/- 0.4 mg/kg. The results demonstrated that both treatments were associated with significant improvements in ADHD symptoms from baseline; however, patients who received OROS MPH demonstrated significantly greater improvements in total ADHD symptoms, inattentiveness and global improvement. The incidence of adverse events was similar in both treatment groups. OROS MPH and atomoxetine are effective and tolerable in the treatment of African Americans with ADHD, and significantly greater treatment responses were observed in patients receiving OROS MPH compared with those receiving atomoxetine over three weeks. Additional studies are needed to evaluate treatment response in this population. PMID- 16350602 TI - Considering cultural diversity in the management of ADHD in Hispanic patients. AB - Attention-deficit/hyperactivity disorder (ADHD) is diagnosed less often in Hispanics than in Caucasian patients. Furthermore, Hispanic patients with ADHD are undertreated. The reasons for these disparities are unknown, although difficulties with access to care among this population may offer a partial explanation. In order to improve treatment outcomes in Hispanic patients with ADHD, healthcare providers must reflect on the diversity of the Hispanic population, which consists largely of persons of Mexican, Puerto Rican and Cuban descent. In spite of the fact that Hispanics share a similar language, religion and belief system, there are other significant cultural differences among these subgroups. In addition, English-language proficiency and socioeconomic variables, factors that are known to influence healthcare outcomes, may also differ markedly among Hispanic subgroups. Therefore, strategies to improve the treatment of ADHD in the Hispanic population must include overcoming language barriers by increasing the availability of Spanish-speaking professionals and medical translators and using culturally sensitive diagnostic instrumentation. Furthermore, improving knowledge of cultural practices of particular Hispanic subgroups may improve the therapeutic relationship between patients and clinicians, facilitate the diagnosis of ADHD and allow healthcare providers to make appropriate treatment recommendations. PMID- 16350603 TI - Men are more accurate than women in aiming at targets in both near space and extrapersonal space. AB - Men excel at motor tasks requiring aiming accuracy whereas women excel at different tasks requiring fine motor skill. However, these tasks are confounded with proximity to the body, as fine motor tasks are performed proximally and aiming tasks are directed at distal targets. As such, it is not known whether the male advantage on tasks requiring aiming accuracy is because men have better aim or is better in the proximal domain in which the task is usually presented. 18 men (M age = 20.6 yr., SD = 3.0) and 20 women (M age = 18.7 yr., SD = 0.9) performed 2 tasks of extrapersonal aiming accuracy (>2 m away), 2 tasks of aiming accuracy performed in near space (< 1 m from them), and a task of fine motor skill. Men outperformed women on both the extrapersonal aiming tasks, and women outperformed men on the task of fine motor skill. However, a male advantage was observed for one of the aiming tasks performed in near space, suggesting that the male advantage for aiming accuracy does not result from proximity. PMID- 16350604 TI - Visual behavior and perception of trajectories of moving objects with visual occlusion. AB - Experienced athletes in sports with moving objects have shown greater skill when using visual information to anticipate the direction of a moving object than nonexperienced athletes of those sports. Studies have shown that expert athletes are more effective than novices in occlusion situations in the first stages of the sports sequence. In this study, 12 athletes with different competitive experience in sports with moving objects viewed a sequence of tennis ball launches with and without visual occlusion, launched by a ball-shooting machine toward different areas with respect to the participant's position. The relation among visual behavior, occlusion time, and the precision of the task is reviewed. The spot where the balls bounced was analysed by a digital camera and visual behavior by an Eye Tracking System. Analysis showed that the nonexperienced athletes made significantly more errors and were more variable in visual occlusion conditions. Participants had a stable visual search strategy. PMID- 16350605 TI - Birth month and suicide among major league baseball players. AB - We examined the association between month of birth and completed suicides among major league baseball players (N = 76). More than twice as many players born in August committed suicide (N = 19) than any other birth month. This association was statistically significant when birth month was corrected for differences in days of the month. The differences were even larger when expressed in terms of birth month for those committing suicides divided by number of major league players born in each month. The association between birth month and suicide raises the possibility that prenatal factors may influence the inclination for suicide later in life. PMID- 16350606 TI - Psychometric testing and applications of the Body Attitudes Questionnaire translated into Portuguese. AB - Although many body attitudes scales have been developed, none have been translated into Portuguese. This study aimed to translate the Ben-Tovim and Walker Body Attitudes Questionnaire and assess validity and reliability for a Brazilian sample. Women with (n = 39) and without (n = 57) eating disorders completed the scale, to assess discriminant validity. Convergent validity was assessed by the correlations between the scores and a measure of body dissatisfaction. The questionnaire was applied twice to controls after 1 mo. to evaluate reliability. Eating-disordered women had significantly higher mean scores on Feeling Fat, Disparagement, Salience and Lower Body Fatness, and lower scores on Attractiveness. No mean difference on Strength and Fitness was found. Most subscales were significantly correlated with the body-dissatisfaction measure. Test-retest coefficients ranged from .57 to .85. The translated version of the Body Attitudes Questionnaire seems to be a valid and reliable measure of body attitudes. PMID- 16350607 TI - Israelis' attitudes toward the disengagement plan, perceived risk, and knowledge of Biblical events. AB - A sample of 73 Israeli students perceived current risks related to the disengagement plan but showed low awareness of Biblical civil wars and political assassinations. Attitude toward disengagement did not correlate with current perceived risks or awareness of Biblical violence. Left-leaning subjects were more supportive of disengagement and perceived greater risk of political assassination in its wake. PMID- 16350608 TI - The Sport Motivation Scale for Children: preliminary analysis in physical education classes. AB - This study was done to test the psychometric properties of the modified version of the Sport Motivation Scale adapted for children in physical education. Participants were elementary school students (N = 452, M(age) = 13.9 +/- 1.04) who responded to the Sport Motivation Scale for Children. The scale assesses three types of motivation at the contextual level, namely, Intrinsic Motivation, Extrinsic Motivation, and Amotivation. Results supported the construct validity (CFI = .95), and internal consistency of the scale (Cronbach alpha > .65). Correlations indicated Sport Motivation Scale for Children simplex pattern exhibiting higher correlations among adjacent subscales than subscales farther apart. The concurrent validity, examined through correlations with scores on the Physical Self-description Questionnaire was satisfactory. Sex differences were examined to assess the discriminant validity. Boys were more intrinsically motivated than girls. Overall, the scale seems a useful one for assessment of motivation in physical education. PMID- 16350609 TI - Cognitive style and plebe turnover at the U.S. Naval Academy. AB - Students entering (N = 1,134) the U.S. Naval Academy class of 2000 were administered the Kirton Adaption-Innovation Inventory on the first day of Plebe Summer, a 7-wk. nonacademic training program completed by all entering students in the summer prior to the freshman year. The mean score on Innovation cognitive style for this sample of plebes was approximately a standard deviation lower than those of five other undergraduate student samples from nonmilitary universities. Furthermore, the 98 plebes who voluntarily withdrew before completing the program scored higher on the average on Innovation than those who remained. Findings suggest that, in terms of Person-Organization fit, plebes with a more innovative cognitive style may, perhaps, be less compatible with the regimentation-style climate of the Academy than those with an Adaptive cognitive style. Further research, however, is needed to specify the relationship between Academy students' cognitive style and other important organizational outcomes. PMID- 16350610 TI - A case study of mediated learning, delayed auditory feedback, and motor repatterning to reduce stuttering. AB - A case study of DW, an 11-yr. old monolingual, English-speaking boy who exhibits stuttering, language delay, and ADHD is presented. DW experienced only limited improvement during stuttering therapy received in public schools, according to parents and the public school clinician. The purpose of this case study was to assess whether fluency treatment which incorporated Mediated Learning, Delayed Auditory Feedback, and Speech Motor Repatterning would enhance progress. Therapy was delivered in two treatments, with each treatment being 5 wk. of intense therapy, separated by one year. Treatment 1 of combined Mediated Learning and Delayed Auditory Feedback yielded improvement in fluency, judged by parents and the teacher to be clinically significant. The improved fluency was maintained for one year when DW was pretested for participation in Treatment 2, which combined Mediated Learning, Delayed Auditory Feedback, and Speech Motor Repatterning Exercises. As no conclusions are possible, further study is needed. PMID- 16350611 TI - Diagnosing children's writing disabilities: different tests give different results. AB - Writing problems are common in children with clinical disorders. However, there are significant intra-individual differences between the ability to spell words, construct sentences, and compose text. Therefore, achievement tests measuring different writing skills may not be consistent in identifying children who have these various writing disabilities. Our study compared scores on the Woodcock Johnson Written Language subtests (which measure the ability to produce single words and single sentences) with scores on the Wechsler Individual Achievement Test Written Expression subtest (which assesses compositional writing skills) in 54 children referred by their teachers to a school psychologist for writing problems. The Woodcock-Johnson only identified 35% of children as having significant writing problems, whereas the Wechsler test identified 78%. Our study suggests that the latter is more likely than the Woodcock-Johnson to identify students who have problems in compositional writing. PMID- 16350612 TI - Psychometric evaluation of the Greek Physical Self-Description Questionniare. AB - To estimate the psychometric properties of the Physical Self-description Questionnaire for deaf high school students, a Greek version was administered to 125 deaf soccer layers and 108 nonexercisers. Confirmatory factor analysis showed adequate factorial structure for the original 11-factor model. Internal consistency and temporal stability were satisfactory for this sample. Also, the construct, discriminant, and predictive validity indices were adequate. These findings supported the use of the Greek version of the Physical Self-description Questionnaire for the assessment of the self-concept of deaf high school students. PMID- 16350613 TI - Relationship between directionality and orientation in drawings by young children and adults. AB - The present study examined the relationship between directionality of drawing movements and the orientation of drawn products in right-handed adults and young children for 27 Japanese kindergartners and 29 Japanese university students who were asked to draw with each hand fishes in side view and circles from several starting points. Significant values of chi2 for distributions of frequencies of orientation of the fish drawings and the direction of circular drawing movement indicated that adult right-handers drawing the fish facing to the left tended to draw a circle clock-wise when they drew with the dominant hand, while there was no such significant relationship in young children's drawings. This result may suggest that the reading and writing habits may be implicated in the direction of drawing movements with the dominant hand, and this directional bias of drawing movement in the dominant hand can appear in the orientation of finished drawings. PMID- 16350614 TI - Accuracy and speed of orthographic processing in persons with developmental dyslexia. AB - A group of 39 persons (20 male and 19 female, 11.0 to 32.5 yr.) with developmental dyslexia and 42 controls (21 male and 21 female, 11.2 to 32.3 years) were compared on computerized tests of sight word reading, nonword decoding, and spelling recognition. The subjects with developmental dyslexia performed significantly slower and less accurately than controls on all tasks. Further, the effect size of the group differences was larger for the older group. Within-group analyses showed a significant difference by age group on accuracy. Only the control group showed a significant age difference between groups on response time. Mean accuracy and response times for the reading-disabled subjects resembled shifted versions of the control group means. These results agree with previous reports that phonological deficits persist for reading-disabled adults and suggest a test of whether the discrepancy between reading-disabled and typically achieving readers may actually increase across age groups. PMID- 16350615 TI - Role of adolescent female volleyball players' psychophysiological properties and body build in performance of different elements of the game. AB - Body-build peculiarities (49 body measurements) and psychophysiological properties (21 computerized tests) of 32 adolescent female volleyballers were studied to assess their significance in performance at competitions. Games were recorded by the original computer program Game, and an index of proficiency representative elements of the game was calculated for each player. Regression analysis was applied to predict the best psychophysiological and anthropometric models for serve, reception, block, feint, and attack. Seven tests of psychophysiological indices and 14 anthropometric variables explained 38-98% and 32-83% of skill performance, respectively. Consequently, to improve young volleyball players' performance, everyday coaching should be complemented by detailed assessment of their body build and psychophysiological characteristics. PMID- 16350616 TI - Summer camp and self-esteem of school-age inner-city children. AB - The present study was designed to test the hypothesis that a session of summer camp would increase the self-esteem of economically disadvantaged, school-age children from New York's inner-city neighborhoods. This study was conducted at a small, coeducational residential summer camp in the Pocono Mountains designed for children ages 6-12 years from low-income areas of New York City. During each of four 12-day sessions, the Piers-Harris Children's Self-concept Scale was administered as a pretest and posttest to a sample of 68 children (36 boys and 32 girls; 33 African American, 34 Hispanic, and 1 Asian) of 742 attending camp for the sumnmer. Children scored significantly higher on the measure of self-esteem at the end of camp than at the beginning. Positive descriptions and ratings of self on popularity increased significantly. Observations and interviews with children suggested physical and social environmental features, such as contact with nature and having the same counselor as a previous year, may support self esteem. Findings are discussed within a framework for biophilia, an innate urge to affiliate with nature which unfolds from earliest childhood on. PMID- 16350617 TI - New measure o f perceived motorcompetence for children ages 4 to 6 years. AB - The purpose of this study was to develop and validate a test for the assessment of perceived motor competence in young children ages 4 to 6 years old. The structure of the Children's Perception of Motor Competence Scale was analyzed through exploratory and confirmatory factor analysis with 495 children between 4 and 6 years of age. A second-order model was selected and consisted of one scale of Perceived General Motor Competence and two subscales, Perceived Gross Motor Competence and Perceived Fine Motor Competence. The number of items was 22. This test showed acceptable internal reliability: global scale (alpha = .81), Perceived Gross Motor Competence (alpha = .80), and Perceived Fine Motor Competence (alpha = .65). Children manifested accuracy in the assessment of the competence. Perceived motor competence was related to actual motor competence as measured by the Movement ABC Test and by an observational scale used by Physical Education teachers. There were no sex differences. The Children's Perception of Motor Competence Scale could be considered an interesting assessment test for identifying current self-perceptions of motor competence in young children. PMID- 16350619 TI - Socialization and behavioral problems of elementary school pupils with developmental coordination disorder. AB - The present study examined the role of the developmental coordination disorder in 154 children's socialization and the expression of deviant behaviors in the context of Greek primary education. For assessment of their motor coordination, the Movement Assessment Battery for Children of Henderson and Sugden was used. The peer nomination method (sociogram) was used for the estimation of children's social status, and the expression of deviant behaviors was assessed via Conners' Teacher Questionnaire. Analyses showed that developmental coordination disorder was associated with poor socialization and the expression of deviant behaviors. These findings support the development of educational programs to include children with poor motor coordination. PMID- 16350618 TI - Spatial- versus object-oriented architectural environments: preference and perception. AB - This study examined changes in perceptual response and preference a specific category of architectural environment by age groups. Understanding relationships among age, perception, and preference is important in creating more appropriate environments. The hypothesis was that individuals who score field-dependent on an Embedded Figures Test would prefer architectural environments which support spatial orientation, while individuals who are field-independent would prefer environments without information on spatial organization or an object-oriented space. It was also hypothesized that children and elderly persons would score more field-dependent and prefer spatially orienting spaces. 64 subjects ages 4 to 85 years were recruited from local schools, various organizations for seniors and adults, given the Embedded Figures Test to measure field dependence, and were compared. Their scores were compared by preference for one of the two environments. Hypotheses were not supported for the spaces overall, however, clear patterns regarding seating preference were identified. Children and seniors had significantly more field-dependent preferences. PMID- 16350620 TI - Measuring men's response to an imagined negative body image. AB - To investigate response to an imagined negative body image, a sample of 29 men from an undergraduate university were given the Body Image Affect Scale of Campbell and Chow. Analysis showed more men, if they are experiencing a poor appearance, would try to cover up physical flaws and avoid approaching an individual in whom they were interested. PMID- 16350621 TI - Evaluating effectiveness of an infusion learning model on attitudes of physical education majors. AB - The purpose was to evaluate the effect of an infusion curriculum model on attitudes of physical education majors toward teaching students with disabilities. 91 undergraduate students (61 men, 30 women) matriculating through the core curriculum of physical education teachers were exposed to the infusion curriculum as part of each course during one academic year. Attitudes were assessed pre- (Fall) and posttest (Spring) on the Physical Educator's Attitude Toward Teaching Individuals with Disabilities III. Analysis indicated a significant positive change in attitudes toward teaching students with disabilities. Findings suggest, when exposed to issues of disability by infusion into course content, these students self-reported they would have increased confidence in teaching and an appreciation when teaching students with disabilities. An independent validation of this change would be valuable. PMID- 16350622 TI - Validation study on self-reported height, weight, and blood pressure. AB - Self-report data were compared with actual recorded data on height, weight, and blood pressure during health examinations to check for validity. The subjects were 1,823 persons, ages 40-68 years. Pearson correlations for systolic blood pressure, diastolic blood pressure, height, weight, and Body Mass Index between data reported in 1993 and measurements made in 1991 were .63, .60, .96, .96, and .92, respectively. Pearson correlations for the same variables between the data reported in 1993 and the measurements made in 1992 were .73, .67, .97, .97, and .94, respectively. Self-reported values for height, weight, and Body Mass Index appear to be valid over years or so. PMID- 16350623 TI - Rethinking of the heuristic-analytic dual process theory: a comment on Wada and Nittono (2004) and the reasoning process in the Wason selection task. AB - This paper raises some methodological problems in the dual process explanation provided by Wada and Nittono for their 2004 results using the Wason selection task. We maintain that the Nittono rethinking approach is weak and that it should be refined to grasp better the evidence of analytic processes. PMID- 16350624 TI - Observation of action and autonomic nervous system responses. AB - Observing somebody performing an action has been shown to elicit neuronal activity in the premotor cortex. This paper investigated physiological effect of observing an effortful action at the peripheral level. As Autonomic Nervous System responses reflect central nervous system processes such as movement planning and programming, it was expected that observing an action would elicit a pattern of ANS responses matching those recorded during actual movement. 12 male subjects, ages 23 to 28 years (M = 25.5, SD = 1.9), were selected as they were experienced in weight lifting. They were asked to observe a squat movement followed by returning to the upright position under 3 different conditions: (i) observation of actual movement performed by somebody else, (ii) observation of a video of the subject himself (first-person video), and (iii) observation of a video of somebody else performing the same movement (third-person video). Moreover, each movement was observed when performed at 50% and 90% of each participant's personal best mark (% of the highest weight which could be lifted). Three ANS parameters were continuously recorded: skin resistance, temperature and heart rate. ANS responses varied as a function of movement intensity: autonomic responses recorded during movement observation at 90% were significantly higher and longer than those recorded during movement observation at 50%. Thus, autonomic responses were linked to the amount of observed effort. Conversely, no difference was found among the three conditions of observation. ANS responses from observation of actual movement were shown to resemble those recorded under the two conditions of video observation. PMID- 16350625 TI - Mental imagery combined with physical practice of approach shots for golf beginners. AB - Recent research on motor skills of golf have pointed to the usefulness of mental imagery. In golf, such training is rarely used as a teaching technique for beginners on the grounds that only top professionals stand to gain from mental imagery. This study tested whether mental imagery combined with physical practice can improve golf performance for the approach shot. 23 volunteer beginners, 8 women and 15 men, M age 23.4 yr. (SD = 3.7), enrolled in the University Physical and Sporting Activities Department, were divided into three groups, using a combination of physical practice of the approach shot plus mental imagery, physical practice only, and a third group engaging in various sporting activities instead of either mental or physical practice of the chip shot. Analysis showed that the beginners' approach shot performance improved most in the group combining physical practice and mental imagery when compared with the group just physically practising the approach shot. It seems mental training can be used effectively to improve performance even with beginners. PMID- 16350626 TI - Assessing a microswitch-based stimulation procedure for eye-blinking responses in a young woman with profound multiple disabilities. AB - We applied a new microswitch-based stimulation procedure for eye-blinking responses with a young woman with profound multiple disabilities, and compared effects of this procedure on the eye-blinking responses and smiling with the effects of a caregiver-based stimulation condition. Analysis showed that the microswitch-based stimulation procedure, with stimulation occurring contingent on eye-blinking responses, increased the frequencies of these responses significantly above the levels recorded in baseline and caregiver-based stimulation conditions. No changes in smiling frequencies occurred. Implications of the findings in terms of alertness, learning, and quality of life are discussed. PMID- 16350627 TI - Role of orientation in perception of emotions. AB - Our study examined whether perception of novel emotions, as with perception of novel objects, elicits a cardiac orientation reaction. Using a habituation dishabituation paradigm, data from 11 adult subjects showed that orientation to both novel emotions and novel objects elicited a heart-rate deceleration. Results suggest that the orientation reaction may be an integral part of perception of emotion. Perception of emotions, therefore, is a complex, multistep process that includes an early orientation reaction. PMID- 16350628 TI - Fitting a psychometric function to price judgments: an analysis of consumers' discounted prices. AB - We examined a psychometric function of price judgment, comparing the mental ruler model, which predicted inverted S-shaped psychometric function, with a power function. Subjects, 102 citizens living in Japan, were asked to rate satisfaction with specified discounted prices. Analysis indicated that predictions of the mental ruler model gave a better fit than those of the power function. PMID- 16350629 TI - Importance of facial physical attractiveness of audiovisual models in descriptions and preferences of children and adolescents. AB - We performed a cross-sectional study with three age groups (8, 14, and 17 years) to evaluate developmental differences in stereotyped beliefs about physical attractiveness and the value of this as perceived by the participants. Given the current importance of television in the development of social knowledge, television models were used. The children and adolescents were asked to evaluate, using bipolar open scales, the physical attractiveness, likeableness, generosity, intelligence, fun, and altruism of 12 television models of both sexes, previously selected by judges, as well as the desire to resemble or feel close to the models. Analysis showed developmental differences across age groups both in the concept of physical attractiveness and in stereotyped beliefs about this. As in other areas of social knowledge, the younger children's responses were bipolar, global, and much more stereotyped, while the adolescents introduced subtle distinctions and elaborated their responses. Nevertheless, physical attractiveness appeared a desirable characteristic for all age groups. PMID- 16350630 TI - Effect of a novel motion desensitization training regime and controlled breathing on habituation to motion sickness. AB - To study whether controlled breathing, known to ameliorate motion sickness, speeds habituation to nauseogenic motion when used in a novel accelerated training regime, 30 participants (13 men, 17 women, ages 18-51 years) were randomized to either a Breathing group or a Control group. Randomization was balanced for rotation tolerance measured during a first exposure to off-vertical axis rotation (OVAR 72 degrees sec.(-1)). Participants subsequently received four exposures to OVAR in 1-hr. (accelerated training). The Breathing group followed taped instructions to control their breathing during training and when retested the next day. There was overall habituation with stimulus repetition, shown by an increase in tolerance for motion, a reduction in symptoms, and speeded recovery times on retesting. There was a tendency for greater habituation with controlled breathing. PMID- 16350631 TI - Preferred body type of fitness instructors among university students in exercise classes. AB - This study examined the relationship between participants' physique anxiety, eating behaviors, somatotype, and the preferred qualifications and body type of a fitness instructor. Subjects were 171 students attending fitness classes at a university. Analysis indicated that the women preferred a thinner fitness instructor than men and that enthusiasm, motivation, and being physically fit were considered the most important characteristics overall. No significant racial differences in key variables were found. PMID- 16350632 TI - Explanation for neon color effect in achromatic line segments on chromatic inducers based on the multiple interpretation hypothesis. AB - In van Tuijl's neon configurations, an achromatic line segment on a blue inducer produces yellowish illusory color in the illusory area. This illusion has been explained based on the idea of the complementary color induced by the blue inducer. However, it is proposed here that this illusion can be also explained by introducing the assumption that the visual system unconsciously interprets an achromatic color as information that is constituted by transparent and nontransparent colors. If this explanation is correct, not only this illusion, but also the simultaneous color contrast illusion can be explained without using the idea of the complementary color induction. PMID- 16350633 TI - Testing assumptions of deliberate practice theory, relevance, effort, and inherent enjoyment of practice on a novel task. AB - This study examined three assumptions of the theory of deliberate practice: that deliberate practice is perceived as relevant for improving performance and that it requires effort, but that it is not perceived as being inherently enjoyable. Of particular interest was the assumption of inherent enjoyment of practice since that has been questioned in previous research. 30 undergraduate and graduate students combined practiced a maze memorization and recall task and rated relevance of practice for improving recall, the practice effort, and inherent enjoyment of practice. The findings were consistent with three assumptions, but also suggested that perceptions about relevance, effort, and inherent enjoyment of practice may change with increasing experience and performance. PMID- 16350634 TI - An 8-item test of acrobatics and balance. AB - A 14-item test of Acrobatics and Balance was previously developed to assess gross motor skills of high school physical education students. This study examined the factor structure of an 8-item version of the test based on exploratory and confirmatory factor-analytic techniques. The 8-item short-form of the Acrobatics and Balance test was administered to 334 Italian boys, ages 14 to 16 yr. (M = 15.0, SD= .8), during physical education lessons. The sample was then randomly split into two groups of equal size. Data from Group 1 were subjected to principal components analysis followed by oblimin rotation. Two factors emerged without overlapping items, accounting for 65.6% of variance. Factors included items of Acrobatics and Balance, matching the scales of the whole version of the 14-item test. Confirmatory factor analysis on data from Group 2 confirmed the two factor structure of the 8-item test. The test had a Cronbach coefficient alpha of .84. The Balance and Acrobatics scales had Cronbach alphas of .84 and .78 for Group 1 and of .84 and .77 for Group 2. Findings provided preliminary support for the use of the 8-item test, especially when there is a need to shorten assessment time. PMID- 16350635 TI - Elongation and enclosure. AB - Previous work suggested that elongation, horizontal area, and the boundary material might influence impressions of enclosure. This article describes an experiment in which these three factors were crossed. Ratings of open/enclosed by 15 respondents on 16 plazas indicated that the elongation ratio (range: 1:1 up to 1:1.8) had the strongest effect on impressions of enclosure (18% of variance). Horizontal area (12.25 m2 vs 49 m2) area accounted for 3% of variance. The amount of variance attributable to boundary material (plants vs walls) was an order of magnitude smaller (0.3%). PMID- 16350636 TI - Exact and resampling probability values for measures of categorical variation and consensus. AB - When analyzing categorical data, it is often important to assess the magnitude of variation, or consensus, among observations in unordered categories. Utilizing the theory of partitions, exact solutions for five commonly used measures of categorical variation are presented. When the number of partitions is very large, resampling methods provide close approximations to exact probability values. PMID- 16350637 TI - Robustness of two single-item self-esteem measures: cross-validation with a measure of stigma in a sample of psychiatric patients. AB - Robins' Single-item Self-esteem Inventory was compared with a single item from the Coopersmith Self-esteem. Although a new scoring format was used, there was good evidence of cross-validation in 83 current and former psychiatric patients who completed Harvey's adapted measure of stigma felt and experienced by users of mental health services. Scores on the two single-item self-esteem measures correlated .76 (p < .001), .76 and .71 with scores on the longer scales from which they were taken, and .58 and .53, respectively, with Harvey's adapted stigma scale. Complex and perhaps competing models may explain links between felt stigma and poorer self-esteem in users of mental health services. PMID- 16350638 TI - Collaborative model shows early success in battling MRSA. PMID- 16350639 TI - Lean production efforts help save 7.5M dollars in 1 year. AB - System undertakes 150 rapid process improvement workshops. Eliminating various types of waste is one of key foci of lean production approach. Time is main metric of methodology; observers measure processes with stopwatches. PMID- 16350640 TI - QIO helps hospital achieve dramatic results. AB - Facility goes above and beyond recommendations for improvement. QIO helps cardiac surgery center identify opportunities for improvement, evaluates PI plan. Unusual weight gains post-op, improving documentation were key areas targeted. PMID- 16350641 TI - Physician satisfaction helps gain buy-in for QI. AB - Physician satisfaction levels jump from the 75th percentile to the 90th percentile. Support sought for cultural shift at hospital, with emphasis on service excellence. Team recognizes doctors for outstanding performance, seeks their input on new ways to support their efforts. PMID- 16350642 TI - AHRMM task force takes aim at supply expense. PMID- 16350643 TI - JCAHO to look closely at patient handoffs. PMID- 16350645 TI - Confusion rules the day as Medicare Part D begins. PMID- 16350644 TI - FTC report arms PBMs in battle with drugstores. PMID- 16350646 TI - Minimal improvement in quality seen in P4P. PMID- 16350647 TI - Physicians and plans can get along. PMID- 16350648 TI - Health plans by design, not by default. PMID- 16350649 TI - Predictive modeling expands its book of techniques. PMID- 16350650 TI - Addressing the hidden costs of rheumatoid arthritis. PMID- 16350651 TI - A conversation with Jayson Slotnik, MPH, JD. Trying to make biotech attractive to payers. PMID- 16350652 TI - How many more quarters of profits on the way? PMID- 16350653 TI - Recently introduced generics helped PBM control its costs. PMID- 16350654 TI - Is chromium III a new key to effective diabetes management? PMID- 16350655 TI - HMOs more solvent and stable--for now. PMID- 16350656 TI - Modern neuropsychology in France: Jean Lhermitte (1877-1959). PMID- 16350657 TI - Does the left inferior parietal lobule contribute to multiplication facts? AB - We report a single case, who presents with a selective and severe impairment for multiplication and division facts. His ability to retrieve subtraction and addition facts was entirely normal. His brain lesion affected the left superior temporal and to lesser extent in the left middle temporal gyri and the left precentral gyrus extending inferiorly to the pars opercularis of the left frontal lobe. Interestingly, the left supramarginal and angular gyri (SMG/AG) were spared. This finding realised a double dissociation with a previously reported patient, who despite lesions in the SMG/AG did not have a multiplication impairment (van Harskamp et al., 2002). The previously suggested crucial role of the SMG/AG in the retrieval of simple multiplication facts is therefore poorly supported (Cohen et al., 2000; Lee, 2000). PMID- 16350658 TI - Spatial working memory and planning ability: contrasts between schizophrenia and bipolar I disorder. AB - Working memory may be conceptualized as a multi-component system involving the active maintenance and manipulation of stored information in the service of planning/guiding behaviour. Impaired spatial working memory is a robust finding in schizophrenia patients which has been related to an impairment in frontostriatal connectivity. The purpose of this study was to examine the specificity of this impairment by comparing the mnemonic and executive aspects of working memory performance in schizophrenia and bipolar disorder with psychotic features, focusing particularly on the functional dynamics between task components. Twenty-four patients with schizophrenia, 14 patients with bipolar I disorder (manic phase) and 33 healthy control subjects were assessed using the Cambridge Neuropsychological Test Automated Battery (CANTAB): including the spatial working memory (between search errors and strategy scores) spatial span (storage capacity) and spatial planning (Stockings of Cambridge: accuracy and latency) tasks. Both patient groups were impaired on the spatial span task, which requires the maintenance and retrieval of stored information. In contrast, only schizophrenia patients showed a significant deficit in between search errors, which requires both maintenance and manipulation of information in working memory. That is, they exhibited both a mnemonic and an executive dysfunction. Spatial span was particularly important to accurate planning ability in bipolar patients. In contrast, in patients with schizophrenia poor spatial working memory was a significant predictor of planning impairments, consistent with failures in goal selection, evaluation and/or execution. Furthermore, initial planning time was positively correlated with the latency to complete a planning sequence. This pattern of slow cognitive processing in schizophrenia patients only, resembled that reported previously in patients with basal ganglia disorders. These findings are discussed in terms of a possible common disturbance in fronto-parietal circuitry in the two disorders together with a specific disturbance of fronto striatal circuitry in schizophrenia, that is not present in bipolar disorder. PMID- 16350659 TI - The ratio of the 2nd to 4th finger length predicts spatial ability in men but not women. AB - On average men score higher on time-constrained tests of spatial ability than women. Both brain and behaviour are influenced by prenatal and adult exposure to gonadal steroid hormones. In humans the ratio of the 2nd to 4th finger length (2D:4D) is a sexually dimorphic character that is lower in men than women and negatively correlated with testosterone levels. We report three independent studies from Sweden/London, Hungary and Liverpool confirming that 2D:4D is generally larger in women than men, that men obtain higher MRT scores than women, and demonstrating that 2D:4D is negatively correlated with MRT score in men but not women. We argue that this negative correlation between 2D:4D and spatial ability reflects the association between 2D:4D and prenatal, rather than adult, exposure to testosterone and conclude that testosterone exposure influences brain development leading to better performance on male-favouring spatial tasks. PMID- 16350660 TI - Neural correlates of script event knowledge: a neuropsychological study following prefrontal injury. AB - Scripts sequentially link information about daily activities and event knowledge. Patients have difficulty sequencing script events following lesions of the prefrontal cortex while showing intact access to selective aspects of script knowledge. It has been suggested that the sequencing impairment is due to a deficit in an inhibitory gating mechanisms that usually enables selection of an item from competing alternatives. If this is the case, then an inhibitory task should reveal script processing impairments on a script categorization task that is not normally associated with poor performance following prefrontal damage. To test this hypothesis, we administered a simple untimed classification task and a modified Go/NoGo task in which subjects classified events from social and non social activities (e.g., read the menu, order the food) and related semantic items (e.g., menu, order) in terms of whether they belonged to a target activity. Participants were patients with lesions of the prefrontal cortex and matched controls. The results showed that damage to the right orbitofrontal cortex was associated with social item classification errors in the simple untimed classification task. In addition, the damage to the right prefrontal cortex was associated with increased response times to respond correctly to Go trials in the modified Go/NoGo task. The data demonstrate that damage to the right orbitofrontal cortex results in impairment in the accessibility of script and semantic representations of social activities. This impairment is exacerbated by an inefficient inhibitory gating mechanism. PMID- 16350661 TI - The effect of explicit cultural bias on lateral preferences in Tunisia. AB - Hand, eye and foot preferences in Tunisia were examined in relation to age, gender and geographic location. We analyzed 1291 questionnaires from 653 men and 638 women, aged 8 to 74 years. Despite the cultural pressure against the use of the left hand for food-related activities, the overall frequency of left-hand writers (10.9%) was comparable to that found in the Western world. The frequency of left-hand writers was higher for subjects with one left-handed parent, and even higher for subjects with two left-handed parents than for subjects whose parents were right handed. The frequency of left-hand writers dropped to 5.9% in the older age-group; it was found to be higher in Tunis, the largest and most Occidental city, than in other cities, while left-hand eating was lower in the South than in the Center or in the North. The frequency of left-footers also dropped in the older age groups and was higher in Tunis than in other cities. Eye preference, consistency of preferred-hand use, crossed hand-eye laterality, crossed hand-foot laterality and gender-related differences in lateral preferences were all comparable to Western results. These data suggest that lateral preferences are partly influenced by a genetic factor, but that handedness (and to a lesser degree footedness) emerges from the intricate interaction of several factors including genetic and cultural influences. PMID- 16350662 TI - Luria: a unitary view of human brain and mind. AB - Special questions the eminent Russian psychologist and neuropsychologist Aleksandr R. Luria (1902-1977) dealt with in his research regarded the relationship between animal and human brain, child and adult mind, normal and pathological, theory and rehabilitation, clinical and experimental investigation. These issues were integrated in a unitary theory of cerebral and psychological processes, under the influence of both different perspectives active in the first half of the Nineteenth century (psychoanalysis and historical-cultural school, first of all) and the growing contribution of neuropsychological research on brain-injured patients. PMID- 16350663 TI - Category-specific effects in object identification: what is "normal"? AB - Previous research on category-specific effects in subjects with intact brains, have found a disadvantage for the identification of natural objects compared to artefacts. This has been taken to support the explanation that natural object deficits in brain-damaged subjects are nothing more than an exaggeration of a "normal" tendency. More recent investigations have, however, uncovered the opposite normal tendency, an advantage for natural objects (Gerlach, 2001; Laws and Neve, 1999). The present study investigated the possible causes of this discrepancy. Three experiments revealed category-specific disadvantages for natural objects in normal subjects despite using stimulus materials rigorously balanced for a) concept familiarity, b) visual complexity, c) name frequency and d) name length. Furthermore, it was found that the very same set of stimulus materials could lead to natural object advantages as well as natural object disadvantages, depending on whether trials in a matching paradigm were positive or negative. In a naming paradigm, there was no natural object disadvantage when stimulus presentation durations were short. The discrepancy present in the literature can be solved by an account emphasising perceptual strategies instead of invoking a lack of adequate experimental control in the previous studies. PMID- 16350664 TI - "Illusions of normality": a methodological critique of category-specific naming. AB - Category-specific disorders are perhaps the archetypal example of domain specificity--being typically defined by the presence of dissociations between living and nonliving naming ability in people following neurological damage. The methods adopted to quantify naming across categories are therefore pivotal since they provide the criterion for defining whether patients have a category effect and necessarily influence the subsequent direction and the interpretation of testing. This paper highlights a series of methodological concerns relating to how we measure and define category (or any) dissociations. These include the common failure to include control data or the use of control data that is inappropriate e.g. at ceiling, unmatched. A review of past cases shows that the overwhelming majority suffers from these problems and therefore challenges conclusions about the purported empirical demonstrations of dissociations and double dissociations in the category specific literature. This is not a refutation of category deficits, but skepticism about the current existence of any convincing empirical demonstrations of category specific double dissociations. As a potential solution, certain minimal criteria are proposed that might aid with the attempt to document category effects that are more methodologically convincing. PMID- 16350665 TI - An illusory illusion? PMID- 16350666 TI - Illusions of category specificity? PMID- 16350667 TI - On double dissociations, controls and gender: some neglected data about category specificity. PMID- 16350668 TI - Category-specific naming and the importance of normal performance. PMID- 16350669 TI - Category-specificity: what is the question? PMID- 16350670 TI - Further methodological concerns in the study of category-specific identification effects. PMID- 16350671 TI - A re-analysis of a case of category-specific semantic impairment. PMID- 16350672 TI - Double dissociations on the same stimuli. PMID- 16350673 TI - Infective endocarditis--an Indian perspective. PMID- 16350674 TI - Diabetes and heart failure: epidemiology, pathophysiology and management. PMID- 16350675 TI - N-terminal probrain natriuretic peptide as a predictor of short-term outcomes in acute myocardial infarction. AB - BACKGROUND: Risk stratification and prediction of high risk for mortality in patients with acute coronary syndromes is based on clinical evaluation, electrocardiogram, biochemical markers and various risk assessment scores. There is emerging evidence that N-terminal probrain natriuretic peptide possesses several characteristics of an ideal biomarker. In this study we looked into the role of N-terminal probrain natriuretic peptide in risk stratification and prediction of short-term events including mortality in patients presenting with acute coronary syndrome. METHODS AND RESULTS: A total of 120 consecutive patients admitted with a diagnosis of acute myocardial infarction, including both ST elevation myocardial infarction (n=80) and non-ST elevation myocardial infarction (n=40) were enrolled. Serum N-terminal probrain natriuretic peptide was measured using electrochemiluminiscence assay (Roche Diagnostics), on the Elecsys 2010 system. On two-dimentional echocardiography, modified Simpson's technique was used to measure the ejection fraction along with end-systolic volume. Various other demographic variables, echocardiographic parameters and risk scores were also assessed. Follow-up at day 30 included a two-dimentional echocardiographic evaluation and assessment for worsening heart failure, recurrent ischemia, and repeat hospitalization. Death due to cardiovascular cause by 30 days was also noted. The mean value of N-terminal probrain natriuretic peptide for the whole cohort was 2307 +/- 2287 pg/ml (271.4 +/- 269.1 pmol/L). For the purpose of comparative analysis, the median value was determined [1403 pg/ml (165 pmol/L)]. In patients having N-terminal probrain natriuretic peptide above median, the end systolic volume was higher while ejection fraction was significantly lower at baseline (p<0.05). At 30 days follow-up, there was a further decline in ejection fraction from 47.7 +/- 11.4 to 43.9 +/- 9.9 (p<0.05), and clinical outcomes were worse in this group. There was a 5% mortality in the entire study group and all patients who died had N-terminal probrain natriuretic peptide above median. On multivariate logistic regression analysis, N-terminal probrain natriuretic peptide above median (OR=32.79, 95% CI 8.74-123.1, p<0.001) emerged as the strongest predictors of adverse outcomes, including 30-day mortality (p<0.001). CONCLUSIONS: N-terminal probrain natriuretic peptide emerged as a strong prognostic tool across the spectrum of acute myocardial infarction and had the strongest predictive value for short-term adverse outcomes including death. PMID- 16350676 TI - Premature coronary artery disease in North India: an angiography study of 1971 patients. AB - BACKGROUND: South Asians, specially Indians, show increased risk for atherosclerosis and have the highest mortality rates due to coronary artery disease amongst all ethnic groups studied so far. We aimed to find out the differences in clinical-biochemical and angiographic profile of young patients versus older patients with angiographically proven atherosclerotic coronary artery disease. METHODS AND RESULTS: Group I (n=828) consisted of patients with age above 55 years (mean age: 63.15 +/- 5.76 years), group II (n=924, mean age: 49.13 +/- 4.25 years) consisted of patients between age 41-55 years and group III (n=219) consisted of patients with age < or = 40 years (mean age: 37.37 +/- 2.92 years). Among the conventional risk factors, smoking was significantly more frequent in group III, while diabetes mellitus and systemic hypertension were more prevalent in groups II and I. Q wave myocardial infarction was more frequently present in groups II and III. Only about one-third of the entire patient population in the myocardial infarction group received thrombolytic therapy. Total cholesterol, low-density lipoprotein cholesterol, and triglyceride levels were significantly higher in younger patients (groups II and III), while high-density lipoprotein cholesterol was significantly low in whole cohort but more so in older patients. Single vessel involvement was more common in group III, while multi-vessel involvement, diffuse disease and fluoroscopic calcium were more common in groups I and II. CONCLUSIONS: Significant differences were observed in the clinical, biochemical and angiographic profile of young patients with coronary artery disease as compared to elderly patients. The younger cohort had more atherogenic lipid profile, higher prevalence of smoking and more frequent single vessel disease. We observed that total cholesterol/high-density lipoprotein cholesterol ratio was a better predictor of coronary artery disease as compared to individual lipid levels. PMID- 16350677 TI - Role of carotid intima-media thickness in assessment of atherosclerosis: an autopsy study. AB - BACKGROUND: The non-invasive technique of measuring carotid artery intima-media thickness has generated considerable interest as a marker of atherosclerosis, particularly in predicting clinical coronary events and coronary artery disease. In the present study, a postmortem comparative analysis of intima-media thickness of carotid artery with coronary artery atherosclerosis has been carried out. To date no such morphological tissue studies are available from our country. METHODS AND RESULTS: Right and left common carotid arteries with their branches were removed at postmortem in 40 cases with history of diabetes, hypertension or both. Intima-media thickness was measured and compared with coronary artery atherosclerosis. There were 10 control postmortem cases without history of diabetes or hypertension. Common carotid artery and internal carotid artery intima-media thickness were found to be good predictors of coronary events. There was also significant correlation (by Pearson's correlation formula) between the carotid artery intima-media thickness and the percentage of block in the coronary arteries. CONCLUSIONS: Internal carotid artery along with common carotid artery intima-media thickness measurement is a good predictor of coronary artery disease. However, carotid artery intima-media thickness has no bearing on the status of collateral circulation of the coronary arteries. PMID- 16350678 TI - Coronary angiogram and intervention through transulnar approach. AB - BACKGROUND: This study was conducted to assess the safety and feasibility of a transulnar approach in performing diagnostic and interventional percutaneous coronary procedures. METHODS AND RESULTS: In the year 2004, a total of 100 patients underwent diagnostic angiography (n=64) and percutaneous coronary interventions (n=36) through transulnar approach. The patients' age ranged from 40 to 70 years and male to female ratio was 7.3:1. The cases of percutaneous coronary interventions were mostly elective procedures and emergency intervention was done in only 2 patients. The procedure was successful in 95 (95%) patients and unsuccessful in 5 (4 diagnostic and 1 percutaneous coronary intervention). The procedure was done through right ulnar artery in all except one patient in whom it was done through left ulnar artery. The total procedure time ranged between 25-45 min. Among the cases of percutaneous coronary interventions, single vessel angioplasty was performed in 23 (65.7%) patients, double vessel in 11 (31.4%) patients and triple vessel in 1 (3%) patient. All percutaneous coronary intervention patients received aspirin, clopidogrel and intravenous enoxaparin. Glycoprotein IIb/IIIa inhibitors were used in 7 patients. Complications such as local hematoma, ulnar artery perforation and reversible parasthesia occurred in one patient each. CONCLUSIONS: Tansulnar approach is a safe and easy alternative technique to perform diagnostic and therapeutic coronary interventions. PMID- 16350679 TI - Effect of combined supplementation of fish oil with garlic pearls on the serum lipid profile in hypercholesterolemic subjects. AB - BACKGROUND: Elevated total cholesterol, especially low-density lipoprotein has been documented as the leading risk factor for the coronary artery disease among Indians. Studies with fish oil supplementation alone have shown an increase in low-density lipoprotein, thereby enhancing the risk associated with incidence of coronary artery disease in hypercholesterolemic subjects. In view of this, the effect of a combined supplementation of fish oil with garlic pearls on the serum lipid profile of hypercholesterolemic subjects was studied. METHODS AND RESULTS: We administered 600 mg of fish oil with 500 mg of garlic pearls (garlic oil) per day to 16 hypercholesterolemic subjects (age range: 30-60 years) with a total cholesterol above 220 mg/dl for 60 days. The effect of this combined supplementation was compared with that of a control group (16 hypercholesterolemic subjects) without any supplementation. The baseline body height and weight of all the subjects were recorded. Significant reductions were seen in all the lipid parameters (except high-density lipoprotein which was increased) in the test group after 60 days compared to that of the control group. The total cholesterol, low-density lipoprotein, serum triglyceride, very low density lipoprotein, and the total cholesterol: high-density lipoprotein ratio reduced by 20%, 21%, 37%, 36.7%, and 23.4%, respectively, and the high-density lipoprotein increased by 5.1% after 60 days of supplementation. CONCLUSIONS: The co-administration of garlic pearls with fish oil was found to be more effective than placebo in the management of dyslipidemia. PMID- 16350680 TI - Use of Amplatzer duct occluder to close severely hypertensive ducts: utility of transient balloon occlusion. AB - BACKGROUND: The dividing line between a patent ductus arteriosus with severe but reversible pulmonary artery hypertension and an Eisenmenger duct remains elusive. Increasing experience with the Amplatzer duct occluder has made non-surgical closure of even large patent ductus arteriosus simple and safe. However, data on the use of this device in severly hypertensive ducti is sparse. METHODS AND RESULTS: There were six patients (2 males, 4 females), wherein transient ductal occlusion was done with an embolectomy balloon catheter. The post-occlusion hemodynamics along with overall clinical and hemodynamic assessment was used to decide the likelihood of benefit of closure and the choice of the device. In four of six patients of patent ductus arteriosus with severe pulmonary artery hypertension (pulmonary vascular resistance index> 8.0 U/m2) the duct was successfully closed using the Amplatzer duct occluder, while in two an Amplatzer ventricular septal defect occluder was used. CONCLUSIONS: Transient balloon occlusion is quite helpful for assessing patients with patent ductus arteriosus with severe pulmonary hypertension, and short-term non-invasive follow-up has shown this strategy to be safe and useful. PMID- 16350681 TI - A case of lipomatous hypertrophy of right ventricle. AB - Lipomatous hypertrophy is a condition that is being increasingly diagnosed with the advent of newer modalities. A middle aged lady was referred to us with recent onset dyspnea and palpitations. Echocardiography revealed diffuse thickening of the right ventricular free wall and outflow tract. Endomyocardial biopsy revealed this as a lipomatous hypertrophy involving the right ventricle. This is the first reported case of lipomatous hypertrophy involving the right ventricle. PMID- 16350682 TI - Successful radiofrequency catheter ablation of recurrent atrial fibrillation due to left inferior pulmonary vein tachycardia. AB - This report illustrates the case of a young lady evaluated for drug-refractory symptomatic paroxysmal atrial fibrillation. Successful isolation of left inferior pulmonary vein was achieved by segmental ostial ablation and circumferential Lasso mapping catheter. Patient is now free of symptoms and is off all anti arrhythmic drugs. PMID- 16350683 TI - Aneurysm of sinus of Valsalva dissecting into interventricular septum with left ventricular communication. AB - Septal dissection with left ventricular communication is a rare complication of aneurysm of sinus of Valsalva. This report describes a case of aneurysm of sinus of Valsalva with septal dissection, almost in its entirety with left ventricular communication--which is a very rare occurrence. PMID- 16350684 TI - Covered stents deployed for coarctation of aorta with aneurysm. AB - Three patients presented to us with upper extremity hypertension and aortic coarctation. Aortic angiograms and spiral computerized tomography delineated the anatomy at the site of coarctation and the associated small aneurysmal dilation. They were taken up for percutaneous stenting of the coarctation segment with cheathum-platinum covered stents. Post-deployment, there was a significant fall in pullback gradients and exclusion of the aneurysms. PMID- 16350685 TI - Spontaneous dissection of coronary arteries. AB - Spontaneous dissection of coronary arteries is an uncommon entity with varied presentation. It is commoner in young patients, specially females. We present three cases encountered by us in recent past. There were two males and the only female was in her post-partum period. All the three had diverse lines of management based on the angiographic picture, clinical background and myocardium at risk. PMID- 16350686 TI - Transposition of great arteries with aortopulmonary window: an unusual cause of prepared left ventricle at 11 months. AB - In patients with transposition of great arteries, presence of aortopulmonary window is very uncommon and associated with high morbidity and mortality. This report describes the case of an 11-month-old female patient in which aortopulmonary window was restrictive, and protected the patient from developing pulmonary vascular disease. The patient underwent successful arterial switch and repair of aortopulmonary window. PMID- 16350687 TI - Current opinions on usage and regulation of drug-eluting stents in India: results of a nation-wide survey of Indian cardiologists. PMID- 16350688 TI - Depression in heart disease--a plea for help! PMID- 16350689 TI - Mechanical breakdown and intraembolus thrombolysis in massive pulmonary embolism. PMID- 16350690 TI - Increased beat-to beat QT variability in patients with congestive cardiac failure. PMID- 16350692 TI - Epidemiology and pathogenesis of human immunodeficiency virus (HIV) related heart disease: a review. AB - BACKGROUND: There is a clear and growing body of evidence for cardiac dysfunction in a significant portion of patients with HIV disease. An increased number of HIV infected individuals may present with cardiac complications in the future as more patients with this disease survive longer because of modern therapy. Heart involvement in AIDS may be well characterized cardiac disease occurring coincidentally in AIDS patients, a complication of the disease or its treatment or possibly a direct insult to the heart by the HIV itself. METHODS: We reviewed the literature on heart disease in HIV infection and AIDS with particular reference to epidemiology and pathogenetic mechanisms that may play a role in diagnosis, management, and therapy of these complications. The MEDLINE/PUBMED and bibliographic searches forEnglish language studies were used. RESULTS: A variety of potential aetiologies have been postulated in HIV-related heart disease, including myocardial invasion with HIV itself, opportunistic infections, viral infections, autoimmune response to viral infection, drug-related cardiac toxicity, nutritional deficiencies, and prolonged immunosuppression. CONCLUSION: An increased number of HIV-infected individuals present with cardiac complications as chronic viral infection, co-infections, drug therapy, and immunosuppression. Understanding the nature and course of cardiac illness related to HIV infection may allow appropriate monitoring, early intervention and therapy PMID- 16350693 TI - Placenta percreta: a review of literature. AB - BACKGROUND: Placenta percreta is a very rare but serious complication of pregnancy. Diagnosis is difficult. Uterine perforation and invasion into adjacent organs makes life threatening haemmorrhage inevitable. Management is usually based on intraoperative findings from case reports. METHOD: Literature on clinical decisions, diagnostic and treatment modalities were critically reviewed using PUBMED and MEDLINE computerized search. Additional Information was also obtained by cross referencing, texts and journals in the medical library of University of Nigeria. RESULTS: Most of the literature was from developed countries with very little report from our environment. There was lack of comprehensive management plan in most textbooks. CONCLUSION: Rare complications are usually very difficult to manage due to lack of experience. It is thus important to identify the patients at risk, attempt to make antenatal diagnosis and involve experienced obstetricians in management. PMID- 16350694 TI - The acute scrotum: aetiology, management and early outcome-preliminary report. AB - BACKGROUND: Most patients with acute scrotal pain may require urgent exploration. We determined the aetiological factors, treatment and early complications in such patients. METHODS: This was a prospective study of consecutive patients presenting with the acute scrotum at Jos University Teaching Hospital, Nigeria from January 2001 to December 2002. RESULTS: Twenty-nine patients were studied with a mean age of 29.4 years (range 12 days to 80 years). Seventeen (58.6%) were 11 to 30 years old. The clinical diagnostic accuracy for testicular torsion was 75% because out of the 24 patients initially thought to have testicular torsion, 18 (62.1%), four (14.8%) and two (6.9%) had testicular torsion, epididymoorchitis and torsion of appendix testis respectively at exploration. The diagnosis in other patients was intrascrotral abscess in two (6.9%), Fournier's gangrene in one (3.4%) and haematocoele in one (3.4%). Only four (13.9%) patients presented within the first 24 hours of illness (all explored within 6 hours) and 14 (48.3%) in one to five days. Thirteen (44.6%) patients had orchidopexy while seven (24.2%) had orchidectomy as a result of testicular torsion (five) and intrascrotal abscesses (two). Testicular exploration only, with antibiotics was the treatment for patients eventually found to have epididymoorchitis, with good response. Testicular salvage rate was 72% in patients with testicular torsion. Postoperative complications observed in three patients were wound infection (two) and wound dehiscence (two). CONCLUSION: We conclude that testicular torsion is the most common cause of acute scrotum in our environment. Majority of our patients with acute scrotum present to hospital late. Health education of the public and attending physicians is required in order to reduce delay in presentation, improve diagnostic skills, testicular salvage rate and prognosis. PMID- 16350695 TI - Pattern and management approach of diabetic foot disease in a developing country. AB - BACKGROUND: Diabetes mellitus is a common endocrine illness in our environment. Diabetic foot gangrene is its commonest surgical complication and is a health burden in Niger Delta region of Nigeria. The aim of the study was to highlight the demography, bacterial incidence, time frame to development of diabetic foot, reaction to ablative surgery and the use of San Antonio wound classification system forthe treatment decision. METHODS: We retrospectively reviewed 46 diabetic feet in 41 diabetics over a 5 year period, January 1999 to December 2003, using the San Antonio classification system. The case notes of all patients seen in the University of Port Harcourt Teaching Hospital were included in the study. RESULTS: There were 32 males and 9 females with a male female ratio of 3.6:1. The age range was 34 to 90 years with a mean of 56 +/- 12 years. The mean duration between diagnoses of diabetes mellitus and development of foot disease was 13 +/- 5 years. Twenty four patients (59.5%) with stages A or B irrespective of the grade had a chance of limb salvage using appropriate antibiotic, serial wound debridement, regular dressing and skin grafting where necessary. Of the 24, 3 absconded, 5 died and 16 healed and were discharged. The remaining 17 patients (40.5%) were stages C and D and were offered ablative surgery irrespective of the grade. Nine discharged against medical advice, 5 died and 3 were discharged in good condition. CONCLUSION: This is a disease of the older population. Delay in accepting ablative surgery affects prognosis. Outcome in management of diabetic foot disease can be improved by education, early presentation, funding for establishment of specialized diabetic foot clinics and early decisive definitive management. PMID- 16350696 TI - Pattern of spinal cord injury in Maiduguri, North Eastern Nigeria. AB - BACKGROUND: Spinal Cord Injury (SCI) is often associated with lifetime morbidity. There is no data on this injury from this Sub-region. The study examines the pattern of SCI in Maiduguri and its environs with a view to contribute to the emerging national data on the rising incidence of this injury and to highlight the deficiencies in our set up and suggest ways of improving them. METHODS: The case notes of SCI patients managed at the University of Maiduguri Teaching Hospital between 1998 and 2002 were retrospectively analyzed. RESULTS: There were 36 patients, 30 males and 6 females (ratio 5:1) with age range of 13-55 years (mean 34.3 +/- 3 years). Road traffic accident (RTA) accounted for 22 (61.1%), while falls resulted in 9 (25%) of the injuries. The most frequent levels of injury were cervical region 14 (38.9%) and thoracolumbar 10 (27.8%). The commonest complications encountered were urinary tract infections (UTI) and bedsores. All the deaths, 3 (8.3%), occurred in patients with complete cervical spine lesion. CONCLUSION: The high morbidity associated with SCI could be reduced through public enlightment on road safety measures including use of seat belts, prohibition of carrying goods and passengers together in trucks. and establishment of Spinal Centres equipped to function. PMID- 16350697 TI - Intraoperative uterine packing with mops: an effective, but under utilized method of controlling post partum haemorrhage-experience from South Eastern Nigeria. AB - BACKGROUND: Uterine packing is a simple noninvasive technique for controlling postpartum haemorrhage. However, its effectiveness has been underestimated and the procedure underutilized leading to lack of experience and expertise especially in the intraoperative usage of mops. The aim of the study was to re evaluate the significance of intraoperative uterine packing in controlling post partum haemorrhage and preserving menstrual and reproductive functions. METHOD: Twenty cases of intraoperative uterine packing with mops performed by the authors over a three-year period (December 2000-December 2003) at three different hospitals were reviewed. Mops were removed after 24 hours. RESULTS: The mean age of the patients was 28.9 +/- 5.6 years, and the mean parity 3.1 +/- 1.8. The mean estimated blood loss was 1500 + 461.1 millilitres and the mean number of units of blood transfused was 1.45 + 1.2. The commonest indication for surgery was placenta praevia (55%). None needed further treatment. CONCLUSION: In carefully selected cases, intraoperative uterine packing is a simple and effective life saving option in our local environment with peculiar beliefs, lack of equipment and expertise. PMID- 16350698 TI - Knowledge, attitude, and practice of female health professionals regarding cervical cancer and Pap smear. AB - BACKGROUND: The objectives of this paper was to determine what female health professionals knew and felt about cervical cancer and Pap smear and their uptake of Pap smear. METHODS: A questionnaire survey of 144 female health professionals at two referral hospitals with facilities for Pap smear. RESULTS: Nurses constituted 52.8% of respondents followed by laboratory scientists (18.8%), doctors (14.6%) and pharmacists (13.9%). One hundred and thirty two (91.7%) respondents had heard of cervical cancer and 80.6% knew it was associated with vaginal bleeding. Twenty two percent could not list any risk factor A significantly greater proportion of doctors (100%) knew the purpose of a Pap smear compared to 59.2% of nurses, 50% of pharmacisits and 48.1% of laboratory scientists (p = 0.003). Only 13 respondents (9%) had everhad a Pap smear with doctors (p = 0.003) and divorced/separated women (p = 0.005) significantly more likely to have done so. Perceived non-availability of Pap smearservice was the main reason (51.3%) for not having had a Pap smear. However, 30.3% had not thought of it or did not consider themselves at risk of cervical cancer. CONCLUSION: Health professionals themselves need to be properly informed about cervical cancer and Pap smears because of their own needs as women and also to improve their effectiveness in educating and encouraging other women to have Pap smears. Available Pap smear services should be publicized and made more affordable. PMID- 16350699 TI - Seroprevalence of HIV infection among blood donors in Port Harcourt, Nigeria. AB - BACKGROUND: This study was undertaken to determine the seroprevalence of human immunodeficiency virus infection among blood donors living in the Niger Delta area of Nigeria. METHODS: A total of 1500 consecutive donors comprising 1481 males and 19 females were screened for HIV-I and HIV-II antibodies using the World Health Organization (WHO) approved Immunocomb HIV I and II test kit. All initially reactive samples were confirmed using the Genscreen HIV I and II test kit. RESULTS: The overall prevalence of HIV was 1.0%. The highest prevalence occurred in the 20 - 29 years age group (60%), followed by 30 - 39 years age group (21.5%). Commercial remunerated donors had the highest infection burden (1.4%) followed by the family replacement donors (0.7%) (p < 0.05). HIV-I accounted for the more predominant strain (0.8%). CONCLUSION: This study has shown an overall prevalence of 1% among blood donors in the Niger Delta area of Nigeria with the highest prevalence amongst donors in the 20 to 29 years age group. We advocate for the mandatory screening of donor units intended for transfusion and the immediate take off of the national blood transfusion service. Furthermore, there is an urgent need for the health education of the people of the Niger Delta area on preventive measures aimed at arresting the spread of HIV. PMID- 16350700 TI - Compliance to antiretroviral therapy among AIDS patients in Aminu Kano Teaching Hospital, Kano, Nigeria. AB - BACKGROUND: Non-compliance with antiretroviral treatment has serious consequences for prognosis and may lead to the development of resistant strains of the virus. This study assessed the level of compliance to antiretroviral treatment, identified factors associated with compliance and reasons for non-compliance among AIDS patients in a Teaching Hospital in Northern Nigeria. METHODS: A pre tested structured interview questionnaire was administered on a cross-section of 263 AIDS patients seen at Aminu Kano Teaching Hospital, Kano. Patient's reported consumption of antiretroviral drugs was compared with the Physician's prescription in the seven-day period preceding the interview. RESULTS: Only 142 (54.0%) of the 263 respondents took at least 80% of the antiretroviral drugs prescribed. Sixty-one (23.2%) did not miss any dose of the drugs. Univariate analysis showed that a significantly higher proportion (58.7%) of patients with formal education were compliant compared to 26.3% of those without formal education (chi2 = 13.7 df = 1 P < 0.01). The educated patient was four times more likely to be compliant [O.R = 3.97, 95% CI = 1.75-9.24]. In contrast, age and sex variables had no significant influence on compliance among these patients. Main reasons for non-adherence to medication include non-availability of drugs (40.6%), forgetfulness (23.9%) and lack of funds (15.8%). CONCLUSIONS: The average compliance observed in this study could be improved by ensuring a steady supply of affordable antiretroviral drugs, better patient-provider communication and enhanced social support for these patients. PMID- 16350701 TI - Transverse cardiac diameter in frontal chest radiographs of a normal adult Nigerian population. AB - BACKGROUND: The possibility of racial differences in heart dimensions has not been extensively investigated in our environment. Cardiac diameter is known to correlate with some anthropometric parameters. Our study was aimed at providing age and sex, specific normal standards of cardiac diameter in an adult Nigerian population. The data were correlated with various anthropometric parameters. METHODS: The transverse cardiac diameter of 303 males and females aged between 20 and 93 years were measured in normal frontal chest radiographs. RESULTS: The mean values for the 20-39, 40-59 and the > or = 60 year-old males were 13.3, 13.9 and 13.7cm respectively. The corresponding values for the females were 12.0, 13.4 and 13.6cm. The range was from 10.5 to 16.2cm in males and 9.5 to 15.7cm in females. Males had larger mean transverse cardiac diameter than females. The cardiac diameter showed a significant positive correlation with age, weight, body surface area, body mass index but not with height. The strongest correlation in males and females was with body mass index. The mean cardiac diameter of females in our study tended to be larger than the values for Caucasian females. CONCLUSION: The strong correlation between the cardiac diameter and body mass index suggests that the cardiac diameter is principally dependent on the body build. PMID- 16350702 TI - Childhood epilepsy: knowledge and attitude of primary school teachers in Port Harcourt, Nigeria. AB - BACKGROUND: This study was conducted to determine the knowledge of primary school teachers in Port Harcourt metropolis of epilepsy, their knowledge of the management of an attack of epilepsy and the attitude of these teachers towards epilepsy in children. METHODS: This is a questionnaire-based, cross-sectional study of 118 school teachers from five randomly selected primary schools in Port Harcourt metropolis, Nigeria. RESULTS: Ten percent (12) of the 118 teachers were graded "Good", 45% (54) "Fair" and 43% (52) "Poor" in overall knowledge score. Sixty six teachers (56%) accept applying crude oil on the body as useful in stopping epileptic attacks in children. There was no significant association between overall knowledge score and sex, year of experience as a teacher and experience with a child with epilepsy. Only 10% of the teachers studied were classified as having overall good knowledge of epilepsy. Sixty nine teachers (58.5%) were graded as having good knowledge of cause of epilepsy. Only 38 (32%) disagree that the saliva drooled during an epileptic attack is contagious; one hundred (84.8%) and 65 (55.1%) agree that some childhood illnesses can cause epilepsy and that it runs in families respectively. Overall, 54 teachers (45.8%) had a cumulative score of negative attitude towards epilepsy. Eighty three teachers (73.3%) would want all children with epilepsy put in a special school whilst 57 (48%) agree that children with epilepsy should be withdrawn from schools. The longer the teacher's professional experience, the more the likelihood of positive attitude towards epilepsy but the association did not reach statistically significant level (p = 0.076). Attitude was not statistically associated with sex and educational qualification. CONCLUSIONS: The overall knowledge of primary school teachers in Port Harcourt metropolis of epilepsy and the first-aid management of an epileptic attack is poor. The attitude of these teachers towards epilepsy is negative. Education of the primary school teacher and general public on epilepsy is recommended. PMID- 16350703 TI - Glycaemic control in type 2 diabetics and the mean corpuscular fragility. AB - BACKGROUND: The osmotic effects of hyperglycaemia and glycosylation of haemoglobin and erythrocyte membrane proteins may play important role in the deformability of RBC in the diabetic state. These effects may be exaggerated in poorly controlled diabetes. The study aimed to determine the fasting blood sugar levels (FBS), glycated haemoglobin (HbA1c) and osmotic fragility of red cells (MCF) in diabetics and non-diabetics. METHODS: Fasting blood sugar, glycated haemoglobin and red cell osmotic fragility were determined in seventy-two diabetic subjects aged between 35-70 years and thirty age matched non-diabetic subjects in Calabar, Nigeria using colorimetric methods. RESULTS: The FBS, HbA1c and MCF were significantly (p < 0.01) higher in diabetics than in non-diabetic subjects. The MCF of diabetics with FBS levels > 7.00mmol/l was significantly higher than those with FBS levels < 7.00mmol/l. No significant difference was observed in the MCF between diabetics with poor glycaemic control (HbA1c > 8.0%) and those with good glycaemic control (HbA1c < 8.0%). The MCF of patients who has been suffering from diabetes for less than five years were significantly (p < 0.05) higher than those who have had the disease for more than five years. A positive correlation (p < 0.003, r = 0.340) was observed between the FBS and HbA1c in diabetic subjects. No significant association was seen between the MCF and HbA1c of diabetic subjects of the study. CONCLUSION: Hyperglycaemia alters the membrane properties of the red cells leading to increase osmotic fragility of the red cells. PMID- 16350704 TI - Pattern of childhood deafness in an audiologic centre in Port Harcourt, Nigeria. AB - BACKGROUND: This study was conducted to determine the incidence of hearing impairment in children visiting a private audiology clinic on Port Harcourt, Nigeria, the type and degree of hearing impairment in these children, and to determine if there is any case for a community-based study on childhood hearing impairment in the region. METHODS: This is a retrospective study of all persons up to the age of 15 years seen in an audiologic centre in Port Harcourt Nigeria over a 42 month period from January 2000 to June 2003. Demographic data were extracted from the medical records of each subject. Audiological evaluation was performed using standard Pure Tone Audiometry for subjects above 3 years and free field audiometric techniques for those below 3 years. RESULTS: Sixty-two (48.9%) of the 127 persons 15 years and below who were examined showed evidence of hearing impairment. There were 29 males and 33 females (male:female ratio = 1:1.14). Age range was 2 to 15 years, mean = 8.3 +/- 4.6 years. Out of the 124 ears examined, 121 showed evidence of hearing impairment whilst 3 were normal. Bilateral hearing impairment was found in 59 (95.2%) of the subjects with hearing loss. Profound, severe, moderate and mild bilateral hearing loss were found in 42 (72%), 7 (11.9%), 6 (10.2%) and 4 (6.8%) subjects respectively. Ninety-seven (83.2%) of the 118 ears in subjects with bilateral hearing loss have, at least, severe hearing impairment. About 90% of the subjects with bilateral hearing loss had sensironeural hearing defect in at least one ear. CONCLUSION: Most of the cases of hearing impairment in our study were of profound/severe severity. Majority were of sensorineural type. There is the need for a community-based study to estimate the prevalence of hearing impairment in the study area. PMID- 16350705 TI - Neologisms in medical practice: their potential to be 'useful', 'useless' or 'misleading'. AB - BACKGROUND: The advances in human endeavours have led to changes in technology. New words, neologisms, have been creeping up and are coined with astounding speed. While these terms are fascinating etymologically, their impact needs to be appreciated from a realistic perspective to keep one aware of the collateral changes we are importing into common language, particularly in medical practice. The aim of this paper is to identify some of such neologisms, outline their current use and then discuss their potential to be 'useful' 'useless' or 'misleading'. METHODS: Terminologies in medical practice and information technology have been selected by the authors and their meanings have been analysed. RESULTS: The terminologies of our present interest include: Computer literacy; Surfing the web; Networking; Information technology; Medical informatics; Telemedicine; Capacity building; Problem-based learning; Mentoring; Learning curve; Evidence-based medicine; Impression; Comorbid conditions; Family medicine; Andropause; Surgical site infection; Confidence interval. CONCLUSION: Neologisms are inescapable in the face of rapid advances in knowledge, equipment and information dissemination to and through diverse cultures and languages. In their assimilation, we should be considerate in discarding familiar terms that have served time. PMID- 16350706 TI - Ruptured heterotopic pregnancy: a case report and brief review of the literature. AB - BACKGROUND: Ruptured heterotopic pregnancy as a cause of acute abdomen is not commonly seen. The aim of this case report is to stress the importance of sonographic examination of the pelvic region in all symptomatic patients in the first trimester of pregnancy. METHOD: The case report of a 23-year-old female with a ruptured heterotopic pregnancy is presented together with a brief review of the literature. RESULTS: A 23 year old female presented with signs and symptoms of an acute abdomen. She gave a history of 8 weeks amenorrhoea. Ultrasound scan (USS) showed an intra uterine pregnancy coexisting with a tubal pregnancy. CONCLUSION: All patients with symptoms of abdominal pain and history of amenorrhoea should have a sonographic examination of the pelvic region. PMID- 16350707 TI - Congenital diaphragmatic eventration in a Nigerian child. AB - BACKGROUND: Diaphragmatic eventration is a rare cause of respiratory distress and failure to thrive in children. METHOD/RESULT: A report of a case of diaphragmatic eventration in a four-month-old child is presented with a review of the relevant literature. CONCLUSION: Clinicians should be aware of this diagnosis and avoid misdiagnosing the condition as a diaphragmatic hernia. PMID- 16350708 TI - Sturge-Weber syndrome in a 56 year old woman: a case report. AB - BACKGROUND: Sturge-Weber syndrome is a sporadic phacomatoses with angiomas involving the leptomeninges and skin of the face, typically in the ophthalmic and maxillary distributions of the trigeminal nerve. Glaucoma is a common ocular manifestation. Presentation is typically at birth with a facial angioma. METHOD: Case note of a patient with a diagnosis of Sturge-Weber syndrome was used and the relevant literature reviewed. RESULT: A 56 year old woman with pain, photophobia in the left eye of 6 months duration. She had lost vision in the eye about 20 years earlier and was born with a dark patch on the left side of her face. She had no previous history of convulsion. On examination, she had a port-wine stain involving the left side of her face. Examination of the left eye revealed a visual acuity of no light perception with episcleral haemangioma. There was a relative afferent pupillary defect and fundoscopy revealed a pale pathologically cupped disc with tortuous retinal vessels. The intraocularpressure was elevated. CONCLUSION: Sturge-Weber syndrome is a rare phacomatoses which may present with ocular complications such as glaucoma. If glaucoma is left untreated decreased vision and blindness result. People of any age therefore with port-wine stain in the ophthalmic distribution of the trigeminal nerve should have yearly eye examination and measurement of intraocular pressure, regardless of whether they have symptoms or not. PMID- 16350709 TI - HELLP syndrome: a report of two cases. AB - BACKGROUND: HELLP Syndrome is a major disorder of the second half of pregnancy that is associated with high risk of maternal and neonatal morbidity and mortality. HELLP is an acronym for Haemolysis, Elevated Liver enzyme levels and Low Platelet Count. As an obstetric complication, it is frequently misdiagnosed at the initial presentation. The purpose of this report is to bring to focus this life threatening disorder that could occur in the second half of pregnancy. METHODS/RESULT: Two cases of HELLP Syndrome are reported. Their conditions were diagnosed and managed accordingly. The first case was delivered by caesarean section while the second case had a vaginal delivery following induction of labour. They fully recovered from the illness and were discharged home from hospital. CONCLUSION: HELLP Syndrome, being an uncommon major disorder of the second half of pregnancy, early diagnosis is critical to avert a high morbidity and mortality, which have been reported to be as high as 25%. PMID- 16350710 TI - Succenturiate placenta: a rare variant in Rivers State Nigeria (a report of one case). AB - BACKGROUND: The objective of this paper is to highlight the complications associated with succenturiate placenta in pregnant women. METHODS: A case report of succenturiate placenta. Thorough examination of the placenta grossly and histologically as well as autopsy of the fetus following standard procedure to confirm the gestational age and to ascertain the cause of death was undertaken by the authors. RESULT: The succenturiate lobe was attached to the major placenta by fibrous tissue which was transversed by blood vessels. The visceral surface showed haemorrhagic necrosis and thrombosis and these led to placental insufficiency resulting in fetal death. CONCLUSION: This rare placental abnormality showed that abnormal placental can be responsible for intrauterine fetal death. This can be detected through the use of ultrasonography. PMID- 16350711 TI - Ocular emergency--a case report. AB - BACKGROUND: Ocular emergencies are not uncommon and one of the causes is chemical burns. Irrigation is the major emergency therapeutic measure of such burns and this also has an impact on the prognosis. We hereby present a case report of chemical burns. METHOD/RESULT: The case report of a 33year old factory worker presenting with history of visual loss following splash of cement dust unto his face is hereby presented with literature review. CONCLUSION: We conclude that people working in high-risk industries for chemical injuries should wear protective devices and be given health education as to what they can do in cases of accident. PMID- 16350712 TI - The resident doctor as a change agent. PMID- 16350713 TI - [Hormone refractory prostate cancer and the prevention of the adverse effects of the treatment]. PMID- 16350714 TI - [Administration of 1 alpha-OH vitamin D3 and calcium prevents bone mass loss in patients with advanced prostatic carcinoma after orchidectomy treated with complete androgenic blockade]. AB - Complete androgenic blockade used in the treatment of advanced prostatic carcinoma can be attained by administration of antiandrogens in orchidectomized patients or by combined therapy with LH-RH analogs and antiandrogens. The treatment, however, decreases the influence of both androgens end estrogens on bone tissue and may result in bone mass loss and increased propensity to fractures. The purpose of the study was to determine the influence of complete androgenic blockade on bone mass and skeletal metabolism in men with advanced prostatic carcinoma and to assess whether 1alpha-OH vitamin D3 (1alpha-OHD3) together with calcium supplementation is able to prevent bone mass loss in men treated with complete androgenic blockade. 51 patients with advanced prostatic carcinoma, with skeletal metastases, aged 44 - 86, mean 68 ys were included into a 12-month prospective study. All patients were treated with orchidectomy followed by therapy with flutamide in a dose of 750 mg daily. 26 patients were additionally given 1alpha-OHD3 in a dose of 0.5 microg/d and calcium carbonate in an initial dose of 1 g daily. It was found that the 12-month treatment with complete androgenic blockade resulted in a decrease in bone mineral density (BMD) by 8.1% in the lumbar spine, by 6.3% in the femoral neck and by 3.5% in the total skeleton. Therapy with 1alpha-OHD3 and CaCO3 caused complete inhibition of bone tissue loss in the lumbar spine and resulted in an increase in BMD by 2.2% in femoral neck and by 1.9% in the total skeleton. None of the examined patients experienced any skeletal fractures. In both groups of patients a prompt decrease in serum alkaline phosphatase activity - a marker of osteoblast activity and an increase in fasting urine calcium creatinine ratio indicating acceleration of bone resorption were found. CONCLUSIONS: in patients with advanced prostatic carcinoma treated with complete androgenic blockade acceleration of bone mass loss is observed; treatment with 1alpha-OHD3 and CaCO3 is able to prevent both trabecular and compact bone loss. PMID- 16350715 TI - Polymorphism of VDR gene--the most effective molecular marker of osteoporotic bone fractures risk within postmenopausal women from Wielkopolska region of Poland. AB - The major public health problem which will arise is a frequency of osteoporosis. The first manifestations of this disease are often bone fractures. Identification and evaluation of individual bone fracture risk will be the most effective way of solving the problem. Genetic determination of osteoporosis is unquestionable. The aim of this study is to detect which variants of genotypes lead to illness. We investigated 187 patients with osteoporosis (161 women, 26 men) and 19 healthy subjects. Polymorphisms of the following genes were investigated: OPG, VDR, ESR1, TGFB1 COL1A1, and BMP2. The statistically significant relationship between BMD value and T allele of Taq I VDR gene were found. Genotypes: aa, bb, TT of VDR gene occur more frequently in polish osteoporotic population in Wielkopolska region within patients with higher risk of bone fractures. PMID- 16350716 TI - [Impact of the mass-reductive therapy with orlistat on 25-(OH)-D3 and PTH concentration in sera of obese, menopausal women]. AB - Epidemiological studies suggest a protective influence of obesity against postmenopausal bone loss. Lower risk of osteoporotic fractures was described in obese patients. However there were only a few studies which examined the effect of weight reduction on bone metabolism and results of these studies are controversial. The aim of the study was to evaluate the influence of weight reduction program using Orlistat on bone metabolism in perimenopausal women. Twenty obese women with simple obesity and without concomitant diseases (BMI 37.1 +/- 3.0 kg/m2, mean age 49.8 +/- 4.6 yrs) were enrolled into this study. The control group consisted of 20 healthy women (mean age 53.5 +/- 5.4 yrs, BMI 24.1 +/- 2.2 kg/m2). All patients have participated in a 3-month weight reduction therapy that consisted of: a 1000-1200 kcal/ day balanced diet (daily calcium consumption about 500mg), Orlistat 3 x 120mg a day and regular physical exercises. Before the weight reduction therapy and after 10% reduction of body weight, serum concentrations of PTH, 25-(OH)-D3, total calcium and phosphorus, total cholesterol were assessed. Dual energy x-ray absorptiometry (DEXA method) of lumbar spine and femoral neck, measuring BMD was performed once, after a 3 month weight reduction therapy using Lunar DPXL. All these measurements were performed only once in control subjects. After a 3-month weight reduction program in patients treated with Orlistat the mean weight loss was 11.6 +/- 5.1 kg which is 12.1 +/- 4.78 %. BMI decreased from 37.1 +/- 3.0 kg/m2 at baseline to 32.6 +/- 2.7 kg/m2 post-treatment. The body weight reduction resulted in significant decrease of body fat and total cholesterol concentration. In obese subjects serum concentration of 25-(OH)-D3 was significantly lower and serum concentration of PTH was significantly higher in comparison to healthy controls, both before and after weight reduction therapy. Serum concentration of PTH, 25-(OH)-D3, total calcium and phosphorus did not change significantly after therapy with Orlistat. CONCLUSION: 3-month weight reduction program using Orlistat did not influence significantly bone metabolism. PMID- 16350717 TI - The effects of growth hormone treatment in patients with somatotropin deficiency during their developmental age. AB - INTRODUCTION: In Poland treatment with growth hormone of adolescent patients dates back to 1964. Till 1993 the therapy was conducted in an interrupted manner, depending on the periodic availability of the drug. The data form such forms of therapy suggested that the end height within 3rd centile was achieved only by a portion of treated patients. Since 1995 the growth hormone is used in continuous therapy, which allows to sum up the effects of the therapy, including the growth rate and end height. MATERIAL AND METHODS: A total of 117 children and adolescent of both sexes, aged 4.6 to 18.1 years, with diagnosed somatotropic or multihormonal pituitary insufficiency were included in the study. All of them were treated with growth hormone and had an analysis of growth rate and end height. RESULTS: In the first 6 months of growth hormone treatment the growth rate achieved 10.4 cm/year in boys and 10.0 in girls and showed no correlation with maturation status. In the second half of the year the growth rate declined slightly. During the remainder of the therapy the growth rate markedly declined, and this effect was most notable in girls. In 93% of patients after the end of therapy the final height was no different than the expected height. CONCLUSIONS: 1. The growth rate in first half a year of the treatment was 3 times higher than before the beginning of therapy. 2. In the second half of the first year the growth rate slightly declined. 3. In following years the growth rate declined notably. 4. The final achieved height in most of the patients does not differ from the prognosed height. PMID- 16350718 TI - [Assessment of VEGF and VEGF receptor concentrations in patients with benign and malignant thyroid tumors]. AB - Neoangiogenesis is a significant event in a cascade of growth and progression of solid tumors. Assessment of the tissue expression and measurement of the concentrations of angiogenic and antiangiogenic factors, contributing to this process, in body fluids, can be used not only for an early diagnosis of tumors and their staging but also as an important parameter of treatment efficiency evaluation. The aim of this study is to evaluate the concentrations of crucial angiogenic cytokine VEGF and its soluble receptors in peripheral blood of patients with benign and malignant thyroid tumors. The study comprised 35 patients with thyroid cancer and 10 patients with follicular neoplasm, both diagnosed by means of ultrasound-guided fine-needle aspiration biopsy. For these patients surgical treatment was instituted. The examined angiogenic factors were determined preoperatively and 4 weeks after the surgical procedures. The results were compared with the control group which comprised 10 healthy individuals. Analysing obtained results, we demonstrated high VEGF concentrations and low soluble VEGF receptor concentrations in patients with benign and malignant thyroid tumors. This fact confirms a vital role of VEGF in angiogenesis of thyroid tumors and a hypothetical antiangiogenic activity of its soluble receptors. Disequilibrium of the above-mentioned angiogenic factor concentrations is probably essential for the growth and progression of benign and malignant thyroid tumors. PMID- 16350719 TI - [The influence of hyperthyroidism and glucocorticosteroid treatment on bone metabolism in patients with Graves' disease and ophthalmopathy]. AB - The aim of the study was to assess influence of hyperthyroidism and glucocorticosteroid treatment on changes of bone turnover markers in patients with Graves' disease and thyroid ophthalmopathy (TO). MATERIAL AND METHODS: Three groups of patients were included in the study. Group I was composed of 26 euthyroid Graves' disease patients with TO suitable for steroid treatment. Group II included 14 hyperthyroid Graves' patients without TO treated medically with anti-thyroid drugs. Group III (control group) included 20 healthy volunteers. Levels of the bone formation marker, i.e. bone-specific alkaline phosphatase (BALP) and the bone resorption marker, i.e. deoxypyridinoline (DPD) were measured in the group I before steroid treatment administration, after 3 methylprednisolone i.v. pulses and after completing the oral prednisone treatment. In the group II levels of BALP and DPD were assessed twice: before treatment of hyperthyroidism and after 6 months since euthyroid state had been achieved. In the group III levels of BALP and DPD were measured once in the basal conditions. RESULTS: Mean initial levels of BALP in groups I and II did not differ significantly and were increased when compared to healthy volunteers. In the group I a transient significant decrease in BALP levels after 3 i.v. pulses of methylprednisolone was observed, followed by a significant increase in BALP after completing the oral prednisone therapy. The achievement of euthyroid state in Graves' patients (II) did not influence significantly BALP values. In the group I initial DPD levels were significantly lower than those in group II and higher than those in the control group (III). During steroid treatment of TO (group I) no dynamic changes of DPD levels were observed. The achievement of euthyroid state in group II was accompanied by a significant decrease in DPD levels, which were however than those in the control group. CONCLUSIONS: 1. In hyperthyroid state is associated with the profound stimulation of bone resorption, and to a lesser extent of bone formation. 2. The achievement of euthyroid state causes a rapid inhibition of bone resorption and maintains a compensatory stimulation of bone formation. 3. Glucocorticosteroid treatment with methylprednisolone i.v. pulses and orally administered prednisone do not influence significantly the processes of bone formation and bone resorption. PMID- 16350720 TI - [Gynecomastia--pathogenesis, diagnosis and treatment]. AB - The aim of this review is to present the up-to-date information concerning the prevalence, pathogenesis, diagnosis and treatment of gynecomastia. Gynecomastia is a benign, unilateral or bilateral enlargement of the male breast due to the imbalance between the androgens and estrogens at the breast tissue level. This clinical condition is particularly common in boys during puberty and in aging men. The breast enlargement, especially with accompanying pain can cause serious psychological problem. At the present time there are no generally accepted procedures for the evaluation and treatment of patients with gynecomastia. In the article such recommendations were proposed. There are many studies conducted to find the safe and efficient medical therapy that could ameliorate the quality of life of the patients with gynecomastia. The information on the available treatment options were also presented. PMID- 16350721 TI - [Genetic aspects of polycystic ovary syndrome]. AB - Polycystic ovary syndrome (PCOS) is a common heterogenous endocrine disorder associated with amenorrhoea (or oligomenorrhoea), hyperandrogenism, hirsutism, obesity, insulin resistance, and an approximately 7-fold increased risk of type 2 diabetes mellitus (NIDDM - non-insulin dependent diabetes mellitus). It is a leading cause of female infertility. The prevalence of PCOS among reproductive age women has been estimated at 4%-12%. Familial aggregation of this syndrome is well established. There are also ethnic and racial variations in the prevalence of the syndrome and its symptoms. Multiple biochemical pathways have been implicated in the pathogenesis of PCOS. Several genes from these pathways have been tested include genes involved in steroid hormone biosynthesis and metabolism (StAR, CYP11, CYP17, CYP19 HSD17B1-3, HSD3B1-2), gonadotropin and gonadal hormones action (ACTR1, ACTR2A-B, FS, INHA, INHBA-B, INHC, SHBG, LHCGR, FSHR, MADH4, AR), obesity and energy regulation (MC4R, OB, OBR, POMC, UCP2-3), insulin secretion and action (IGF1, IGF1R, IGFBPI1-3, INS VNTR, IR, INSL, IRS1-2, PPARG) and many others. Most women with PCOS, both obese and lean, have a degree of insulin resistance. The minisatellite of insulin gene (INS VNTR), especially class III alleles and III/III genotypes might not only determine the predisposition to anovulatory PCOS but also the concomitant risk for development of type 2 diabetes. The function of the insulin receptor (IR) is probably normal in woman with PCOS. However abnormal serine phosphorylation in the receptor may impair signal transduction accounting for a post-binding defect in insulin action. Serine phosphorylation is also involved in the postranslational regulation of 17,20-lyase activity (CYP17). There may be a common aetiology for both insulin resistance and hyperandrogenism. Polymorphic alleles of both IRS-1 and IRS-2 (insulin receptor substrate 1 - 2), alone or in combination, may have a functional impact on the insulin-resistant component of PCOS. There is no evidence to suggest that follistatin gene polymorphisms play a role in the pathogenesis of insulin resistance in PCOS women. PCOS appears to be associated with the absence of the four-repeat-units allele in a polymorphic region of pentanucleotide (TTTTA)n repeats within CYP11A gene, which encodes cytochrome P450scc. It has been hypothesized that up-regulation of this enzyme could lead to increased androgen production. There is no evidence of any association of alleles of CYP19 gene (encoding cytochrome P450arom) with PCOS. Association exists between androgen receptor gene (AR) polymorphisms an androgens action in PCOS. Increased hirustism and decreased CAG repeat length within AR gene has been also demonstrated in women with normal testosterone levels. Expression of estrogen receptor (ERs) as well as 5-alpha-reeducates (SRD5A1-2 genes) activity was analysed in granulosa (GC) and theca cells (TC). The results of this study demonstrate that there are significant alterations in the expression of ERalpha and ERbeta in PCOS that may be related to abnormal follicular development. On the other hand elevated SRD5A activity in polycystic ovaries supported the hypothesis that 5-alpha-reduced androgens may play a role in the pathogenesis of the syndrome. The genetic aetiology of PCOS remains unknown. There are a number of interlinking factors that affects expression of PCOS. Single cause of PCOS is unlikely. Other possible mechanisms in pathogenesis of PCOS are discussed. PMID- 16350722 TI - [DNA microarrays and papillary thyroid carcinoma gene expression profile]. AB - The paper presents gene expression profile analysis with DNA microarrays and compares two core technological platforms used for this purpose - high density oligonucleotide microarrays and cDNA microarrays. With this background recent results of papillary thyroid carcinoma analysis with DNA microarrays are presented. PMID- 16350723 TI - [Diagnostic and therapeutic difficulties in MEN 1 syndrome]. AB - MEN 1 syndrome (Multiple Endocrine Neoplasia type 1) is a rare endocrine disorder characterized by the association of tumors in several endocrine glands, mainly in parathyroids, gut and pituitary. At our institution in the years 1982-2004 we have followed 26 patients with MEN 1 syndrome belonging to 19 families. The diagnosis of MEN 1 was based on Gubbio Consensus (JCEM 86: 5658-5671, 2001). Mean age at the diagnosis of MEN 1 was 35 years. Primary hyperparathyroidism was the most frequent pathology, which was diagnosed in 25 of 26 patients (96%). Gut endocrine tumors were found in 20 patients (77%), while pituitary tumors in 18 (70%). Non-functioning gut tumors were most frequent (n=9), followed by insulinoma (n=7) and gastrinoma (n=4). Prolactinoma was the most frequent pituitary tumor found in 12 patients (67%). Three patients died during the observation period - all of them of generalized gut endocrine tumor (gastrinoma in 2 cases and foregut carcinoid in one case). The management of MEN 1 is not easy and careful analysis of clinical picture is necessary in each individual case. Several important observations can be made on the basis of own experience and the literature: 1. In each sporadic pathology, which may be a part of MEN 1, one should consider. the possibility of MEN 1. The individual MEN 1 abnormalities are often diagnosed after 40 and later 2. MEN 1 tumor are usually multiple thus necessitating a different therapeutic approach (more radical surgery) 3. The most valuable screening tests are: Ca++, PP, CgA and prolactin 4. Endoscopic ultrasound is the most specific method for the localization of pancreatic endocrine tumors. 5. The results of surgical treatment of MEN 1 tumors are worse than that of sporadic tumors. 6. Prognosis in MEN 1 is determined by the behaviour of gut neuroendocrine tumor 7. No genotype/phenotype correlation in MEN 1 syndrome was found so far. In summary, it should be underlined that MEN 1 syndrome is an endocrine disorder, in which early diagnosis and optimal treatment may significantly improve the prognosis. PMID- 16350724 TI - [Molecular action of insulin-sensitizing agents]. AB - Atypical endometrial hyperplasia has been associated with progression to endometrial cancer, the most common genital malignancy. There are multiple risk factors for endometrial cancer, such as early menarche, exogenous estrogen exposure, obesity and diabetes. Diabetics have a 3-4 fold relative risk of endometrial cancer. Also, several studies have demonstrated an association between insulin resistance and endometrial cancer. There is known the first description of atypical endometrial hyperplasia resistant to progestogen therapy, which was subsequently treated with an insulin-sensitizng agent, metformin. Metformin is a biguanide antihyperglycemic agent used in the treatment of adult onset diabetes. Unlike the sulfonylureas, metformin does not act primarily by increasing insulin secretion. In contrast, metformin lowers the rate of gluconeogenesis in the presence of insulin. Therefore, it is considered an insulin-sensitizer. Increased insulin sensitivity may improve the metabolic effect of insulin and decrease its mitogenic effect by tissue-specific mechanisms. One explanation for tissue specific differences in insulin binding and action may be through the relative expression of the insulin receptor (IR) isoforms. The IR isoforms IR-A and IR-D differ by 12 amino acid residues, owing to the alternative splicing of exon. The IR-A is predominantly expressed in malignant tissues and may lead to mitogenic effects within the cell. The relative expressions of IR-A and IR-B in normal and malignant endometrial tissue is not known. Besides direct effects on the IR, several additional mechanisms have been proposed for the mitogenic effect of insulin in endometrial cancer. In addition to the possible direct mitogenic effects of insulin through the IR-A, insulin resistance may be associated with alterations in expression of insulin-like growth factors (IGFs) and the IGF binding proteins (IGFBPs) or may inhibit the protective effect of progestagens. Binding sites for IGF-1 and IGF-2 have been confirmed in both normal and malignant endometrium. Binding of IGF-1 is significantly higher in endometrial cancer compared to normal endometrium. In the Ishikawa human endometrial cancer cell line IGF-1 was a more potent mitogen than insulin or IGF-2. Insulin may increase mitogenicity by regulating the expression of IGFBPs. The IGFBPs are a family of proteins that have both proliferative and anti-proliferative effects. While all six high-affinity IGFBPs are expressed in the endometrium, IGFBP-1 is the best characterized. Hyperinsulinemia can decrease IGFBP-1 even in the presence of progesterone, perhaps inhibiting progesterone's protective effects. Interestingly, IGFBP-1 was undetectable or minimally expressed in endometrial cancers. Nestler discussed results of a 6-month treatment of 100 nonebese women with PCOS, which showed a somewhat greater effect of metformin than rosiglitazone and no benefit of administering both agents in combination. Long-term treatment with oral contraceptives decreases endometrial cancer, with a reduction in serum androgens and a decreases in hirsutism and acne, but may worsen insulin resistance and lead to deteriration in glucose tolerance. Insulin sensitizers, on the other hand, should decrease endometrial hyperplasia by inducing regular menses, but may not be as beneficial in improving androgen - related symptoms. Note that the Nurses Health Study (NHS) showed increased risk of diabetes in oral contraceptive users. These considerations may be related to the finding that women who used oral contraceptives have increased risk of myocardial infarction. Thus, in view of the particular increase in CVD risk among women with PCOS, one might be less likely to recommend oral contraceptives, while insulin sensitizers may be of particular benefit, decreasing androgens, improving ovulation and fertility, and reducing the risk of diabetes and CVD. Theoretically, metformin, a treatment which is now widely used to treat infertile women with PCOS, may have a role in preventing endometrial hyperstimulation by lowering insulin concentrations and restoring ovulation. However, the long-term effects of this drug in women with PCOS are not known and more studies are required before suggesting its use for preventing endometrial cancer. PMID- 16350725 TI - [Physiological significance of estrogens in men--breakthrough in endocrinology]. AB - Estradiol (E2) is traditionally recognised as the female sex hormone. It has been believed for 40 years, that E2 didn't exert any influence or caused impairment of the gonadal function in men. The main source of E2 in men is adipose tissue and the brain. E2 is also produced in adrenals, liver, mammary glands, hair and in male gonad. Daily production and the level of E2 in the blood in men are higher than those in postmenopausal women. At the end of the 80-ties we were first reporting that during sexual maturation E2 can be the important hormonal signal for the initiation of spermatogenesis. The traditional view about unimportant or inhibitory role of E2 in male physiology was finally refuted thanks to discovering estrogen receptors in males. In the 90-ties, transgenic mice with the lack of estrogen receptor (ER) or gene encoding enzyme aromatase, that enable the conversion of testosterone into E2, were also produced. Observations of men with inherited mutations of these genes, considerably extended our knowledge about E2 in men in stroma bones formation, inhibition of their growing, lipids metabolism and sexual maturation, the effects that were attributed to testosterone action until today. New data also points at important role of estrogens and ER in the function of the cardio-vascular system and in counteracting coronary disease in men. PMID- 16350726 TI - [Membrane receptors for estradiol--new way of biological action]. AB - Classical action of steroid hormones, called genomic, includes binding to their intracellular receptor, require hours or days to occur and require transcriptional effects with subsequent modulation of protein expression. Some of the biological effects induced by steroids, and mainly by sex steroids, take place within seconds or few minutes, time far too fast to be due to the genomic changes. The rapid, nongenomic action of estradiol are attributed to membrane action, probably through variety of proteins present in cell membrane. The rapid effects of steroid hormones are manifold, ranging from activation of protein and tyrosine kinases, G proteins, and modulation of ion channels. The nongenomic way of action includes also non-direct control of processes of transcription and gene expression. There are at least three different way to interact with cell membrane. Steroids may change membrane fluidity, without binding to any known protein or receptor. Another way is allosteric modulation of non-specific for steroid hormones receptors, or structural and enzymatic protein present in cell membrane. Evidence suggests that the classical steroid receptors can be localized at the plasma membrane, triggering signals typical for G-proteins coupled receptors. Physiological significance of nongenomic action of steroids needs to be elucidated. PMID- 16350727 TI - [Molecular aspects of the etiopathogenesis of the parathyroid gland diseases]. AB - Current views on the molecular aspects of familial parathyroid gland diseases have been presented (familial primary hyperparathyroidism, hypoparathyroidism and psuedohypoparathyroidism). Their inherited mode and genetic abnormalities have been described. Particularly, the following genes: HRPT2, MEN1, RET, CASR, GNAS have been shown. Localization, structure, expression and structural changes (mutations) found in patients with familial parathyroid gland diseases have been presented. Attention has been paid to clinical and histopathologic symptoms, which should indicate the need to undertake genetic studies. PMID- 16350728 TI - [Molecular aspects of pituitary tumors]. AB - Pituitary adenomas are common benign neoplasms, accounting for approximately 15% of intracranial tumors. In systematic autopsy, pituitary tumors are found in 25%, of the population, but only one-third of these tumors give rise to clinical manifestations. Why most of these neoplasms remain undiagnosed and pituitary carcinomas are extremely rare? The progress in the studies concerning pituitary tumorigenesis is rather slow and, due to several limitations, including the anatomic inaccessibility of human pituitary gland, the lack of functional human cell lines in culture and the discrepancies between human and animal pituitary oncogenesis (in rodents pituitary hyperplasia is a prerequisite for adenoma development). In humans, the majority of pituitary tumors are monoclonal in origin and derived from single mutated pituicyte, rarely hyperplasia is a prerequisite for adenoma formation. As in the case of other tumors, activating mutations in oncogenes (GNAS1, PTTG) and inactivating mutations in tumor suppressor genes (MEN1, CNC1) lead to pituitary tumors development. However, mutations in classic oncogenes are very rarely associated with these tumors. Moreover, the important role of some hypothalamic hormones, peripheral hormones and their receptors (e.g. GHRH, dopamine D2 receptor, PRL receptor, estrogens, thyroid hormone receptor) and growth factors (e.g. FGF, EGF, TGF) is postulated and partially proved in promotion of pituitary tumorigenesis. Further studies are required to determine which of these events are truly primary changes in pituitary tumorigenesis, what may allow development of gene therapy. PMID- 16350729 TI - [Genetic factors predisposing to the development of papillary thyroid cancer]. AB - According to classic theory of neogenesis, cancer arises from well-differentiated cell that in response to variety of factors de-differentiates, becomes able to proliferate without control and/or loses its ability to undergo apoptosis. According to another theory, cancers (at least cancers of some organs) originate from stem cells, which "by definition" are poorly differentiated and able to proliferate indefinitely. Therefore a lower number of abnormal events is necessary for these cells to escape proliferation-controlling mechanisms. With regard to papillary thyroid cancers it is still thought that it arises from well differentiated thyreocyte. One of the characteristic features of cancer cell is chromosomal instability. Lowest number of such abnormalities is observed in well differentiated thyroid cancers (including papillary cancer), intermediate - in poorly-differentiated cancers, while highest - in anaplastic cancers. Microarray analysis shows that despite of clinical heterogeneity, gene expression profiles of papillary cancers are very similar. Genetic anomalies predisposing to the development of papillary cancer most commonly regard proteins that possess kinase activity. Kinases phosphorylate other proteins, and play an extremely important role in signal transduction from outside the cell as well as inside the cell. Constitutive activation of some kinases may lead to the excessive and/or permanent activation of some transduction pathways specific for mitogens or growth factors. This results in excessive proliferation. The best known protein of such type which function is altered in papillary thyroid cancers is RET - a membrane-located growth factor-receptor with kinase activity. RET gene undergoes different rearrangements in this type of cancer. There are approximately 10 RET rearrangements known, with RET/PTC3 and RET/PTC1 being most common. In this anomaly kinase domain-encoding 3' end of RET gene is aberrantly bound to 5' end of another gene. Fusion protein synthesized on such hybrid template is not present in the cell membrane but in the cytoplasm, where it permanently activates transduction pathway specific for RET. NTRK1 gene encoding a member of family of neuronal growth factor receptors containing thyrosine kinase domain is also rearranged in papillary cancers. However, genes fused to its kinase domain encoding sequence are different from the ones fused to RET. MET, a gene encoding another membrane protein with thyrosine kinase activity, which acts as a growth factor-receptor, is overexpressed in 70%-90% of papillary thyroid cancers. BRAF gene encoding another yet kinase transducing signals from RAS and RAF to the cell is mutated at position 1796 (T/A, amino acid substitution V599E) in 38-69% of papillary cancers. The presence of this activatory mutation is associated with higher degree of clinical advancement of the disease. In addition, in majority of papillary cancers tested, mutations of the genes encoding nuclear triiodothyronine receptors were found. Transgenic mice with both TRB allele replaced with dominant-negative TRB mutants develop aggressive thyroid cancers. Progression from papillary to anaplastic cancer is most possibly caused by the occurrence of additional anomalies within P53, RAS, NM23,b-catenin gene and other genes. PMID- 16350730 TI - [Diagnostic value of immunohistochemistry in lesions of the endocrine system]. AB - The aim of immunohistochemistry and immunocytochemistry is to reveal specific antigens in cells and tissue samples. Those techniques are based on an antigen antibody reaction and visualization of its product in microscopic examination. The precursor of this new diagnostic procedure was an immunofluorescent reaction in frozen tissue samples performed by Albert Coons in 1940. Then the immunohistochemical techniques were perfected to increase sensitivity and specificity. Currently it is hard to imagine a modern pathological examination without immunohistochemistry. At the end of XXth century it was believed that 75% of cases is possible to be diagnosed due to immunohistochemical stains. Microscopic examination of endocrine glands tissue samples is extremely difficult because of coexistence of the presence of neoplasms and endocrine dysfunction. It is necessary to establish the type of hormones in the cells of the endocrine system lesions to make a proper diagnosis. Thanks to the use of antibodies against hormones and its precursors it becomes possible. At present most of the antigens are easily detected in both: formalin fixed paraffin embedded tissues samples and ethanol fixed cytological smears so immunohistochemocal and immunocytological stains can be a part of routine diagnostic procedures in pathology. However most of the biologically active substances are revealed in many organs and tissues and it is necessary to perform a satisfactory immunohistochemical panel to be sure the diagnosis. It is important to notice that there is no need to make a wide panel of antibodies in all of the cases and the economical aspect of examination is also important. Of course immunohistochemistry sometimes is the guarantee of proper diagnosis but in some cases too wide panel of antibodies can be a loss for the patient and for medical department. We discussed the proper diagnostic procedures and immunohistochemical profile in pathological lesions of endocrine system (thyroid and adrenal gland, adenohypophysis, neuroendocrine tumours and some hormones-secreting tumours of gonads). PMID- 16350732 TI - [Genetic aspects of premature ovarian failure]. AB - Among the causes of premature ovarian failure (POF) two groups of factors are reported: factors which lead to decrease of follicular number and factors which stimulate follicular atresia. In the first group genetic factors are the most important whereas in the second: enzymatic autoimmunological, iatrogenic, toxins and infections are reported. In 1986 familiar POF on the background of long arm of chromosome X deletion was reported. Other chromosomes which are important for normal ovarian function are: chromosome 21 (AIRE gene), chromosome 11 (gene of beta FSH, ATM gene), chromosome 3 (gene responsible for BEPS syndrome) and chromosome 2 (genes of FSH and LH receptors). In this review the role of these genes and results of several epidemiological studies are reported. PMID- 16350731 TI - [The role of oocyte in the genetic determinations of fertility and infertility]. AB - Oocyte specific genes play important role in the foliculogenesis, ovulation, fertilization and early embryogenesis. It is suggested that 17-20% of infertility in both sex has idiopathic aspect. This kind of infertility is mainly associated with genetic background. The study on the role of oocyte specific genes can help in our understanding of the causes of idiopathic infertility. PMID- 16350734 TI - Verify, clarify and confirm. PMID- 16350733 TI - [Medullary thyroid carcinoma: from molecular studies to clinical decision]. AB - The paper is focused on guidelines of practice in inherited medullary thyroid cancer, diagnosed on the basis of DNA analysis. Identification of RET mutation implies further steps of diagnostic procedure, some of them - USG, FNAB and calcitonin level tests - are common for all types of mutation, other are related to ascertained type of mutation. In asymptomatic RET mutation carriers, prophylactic thyroidectomy is indicated. In MEN2B inherited cancer reveals its symptoms quickly and shows dynamic progress. In MEN2A/FMTC the clinical picture is diversified - in some patients the course of disease is mild, however in some other cases the progression of disease and even death occur regardless of the proper treatment. Unfortunately, there are no molecular prognostic markers in medullary thyroid carcinoma. Recent papers and also our own unpublished results show that gene expression profile, is similar in MEN2A and sporadic cancer. This group differs from MEN2B by its expression profile. In conclusion it is to be emphasized that although inherited medullary thyroid carcinoma is a rare disease, the diagnostic algorithm is well established and maximizes the chance for early diagnosis. Moreover, it needs to be stressed that DNA analysis results inform us not only about the necessity of further therapy, but also suggest different ways of proceeding in particular type of mutation. PMID- 16350735 TI - The Nursing and Health Outcomes Project. AB - Increasingly, administrative databases are playing major roles in decisions regarding the effectiveness and funding of the health-care system. These databases contain information taken from patients' charts after their discharge and include their age, gender, diagnoses, medical interventions and length of stay Although nurses are the largest health-care provider group in all sectors of the Canadian health-care system, the nature and effectiveness of their contributions to patient care are not captured on administrative databases because no information about nurses' contribution to care is abstracted from charts. The Nursing and Health Outcomes Project (NHOP) was established by the Ontario Ministry of Health and Long-Term Care to identify nursing-sensitive patient outcomes and their attendant nursing inputs and processes that could be housed on databases. This article describes the background to this project and reports on the progress to date. The implications of having this information on databases will be examined. PMID- 16350736 TI - HIV prevalence in older adults. AB - The "overlooked epidemic," "silent risk group" and "forgotten population" are phrases used when referring to HIV/AIDS and the population group aged 50 and older Health Canada reports that almost 12 per cent of the positive HIV test reports in Canada since the epidemic began are among those 50 and older. Despite the availability of this information, older adults (and those who care for their health) are often not aware of the risk in this group. Older adults are often stereotyped as asexual beings and health-care providers frequently omit a sexual history as part of their assessments. Symptoms of AIDS such as fatigue and memory loss can be dismissed as part of the normal aging process and this may lead to the fact that older people frequently present with AIDS at the same time they are diagnosed with HIV. Research, prevention and education programs are typically geared toward younger adults. Multiple intervention programming must be designed and implemented specifically for older adults in order to reduce HIV transmission in this group. The nursing community is in a strategic position to intervene. The authors discuss various strategies for action. PMID- 16350737 TI - Multidisciplinary Collaborative Primary Maternity Care Project. A national inititative to address the availability and quality of maternity services. PMID- 16350738 TI - A community hospital acute pain service. AB - This article provides readers with a guide to developing and implementing an acute pain service in a community hospital. Kanter's theory of innovative diffusion is used to frame the author's experiences as a lead nurse in two community hospital acute pain services. Health-care providers recognize the importance of quality pain assessment and management. One initiative for improving pain management has been the implementation of an acute pain service (APS). In Canada, most university-affiliated teaching hospitals have now developed an APS to improve pain management. Community hospitals, however, have only recently begun to adopt the concept. Improving pain management through an APS provides an excellent opportunity for nursing leadership at all levels. Nursing administration may take the lead in proposing the idea and benefits of acquiring an APS. An advanced practice nurse can provide leadership through the coordination and provision of enhanced pain management as a lead nurse in an APS. Staff nurses can provide leadership in improving pain management on a daily basis and ensuring that quality pain care reaches the bedside, Nursing practice is at the core of making a difference in pain management. PMID- 16350740 TI - [Analysis of some "insoluble" problems of determining the binding parameters of ligand-receptor interaction and methods of their solving]. AB - Some problems of the estimation of the parameters of ligand-receptor interaction (affinity, rate constants, valency, etc.) were considered. It was demonstrated that not only the Scatchard plot but also Klotz plot could be used for determining the parameters of ligand-receptor interaction for two types of binding sites of different affinity. A new approach and new coordinate systems for the estimation of the parameters of ligand-receptor interaction were suggested. It was shown that for the estimation of the affinity of putative monovalent antibodies by ELISA various equations, which are more precise and convenient than the Friguet et al. equations, could be obtained by the transformation of mass action law equation. The problem solution for the estimation by ELISA the affinity of two types of bivalent antibodies with different affinity and their concentrations for the case of the mixture of these antibodies was also suggested. The application of the proposed coordinate of dilution allows to solve the problem of determination of the parameters of ligand receptor interaction (including antigen-antibody system) for the pre-existing ligand-receptor mixture without their preliminary separation and purification. This approach is especially important for the cases when the receptor is not stable enough to be isolated in the intact form from this mixture. It was shown that the well-known phenomenon of the prozone often observed under the titration of serum antibodies by the method of agglutination may get a mathematical explanation. Analytical solution of the problem of determining the velocity constant and the amount of the end product of the first order irreversible and reversible reaction kinetics was suggested, despite the fact that the process is described by the system of irrational equations. Mehods of asymptotic solution of transcendental irrational equations which describe the dynamics of reactions which mechanisms are subject to the so-called heterogeneous, successive, or competitive models have been considered. These methods permit the finding of the reaction rate constants and the amount of the end product, if the kinetics of the transformation of either initial, or end product of the reaction is known. PMID- 16350741 TI - [Effect of tissue-type plasminogen activator and its inhibitor in haemostasis system function in norm and pathology]. AB - The paper is dedicated to the study of structure and physiological functions of tissue type plasminogen activator and its inhibitor PAI-1, changes of these parametres in normal and pathology conditions. The interrelation of the coagulation and the fibrinolytic system during the range of pathologies was studied. It was demonstrated that simultaneous analysis of the coagulation and fibrinolytic systems analysis allow to diagnose the thrombotic complications. PMID- 16350742 TI - [Substrate specificity of brain cholinesterase activity of evolutionary old species of fishes]. AB - An analysis of brain cholinesterase properties in 3 species of cartilaginous fishes and 1 species of chondrostei, was conducted for the first time. The skates brain enzymes, as to substrate specificity, are shown to be referred to acetylcholinesterases, and the shark brain cholinesterase - to atypical acetylcholinesterase. Kaluga brain cholinesterase, as to its substrate specificity, differes from brain enzymes of cartilaginous fishes. The differences in properties of the nervous tissue cholinestarase of the investigated fish species are the parameters of evolutionary divergence of species. PMID- 16350743 TI - [Kinetic characteristics of Ca2+, Mg2+-ATPases in cells of the submandibular salivary gland of rats]. AB - Kinetic properties of Ca2+, Mg2+-ATPases membranes from acinar cells of rat submandibular salivary glands have been investigated. It was found that kinetics of ATP hydrolysis dependent on Ca2+, Mg2+-ATPases corresponds to the first-order reaction during first 2 min. It was found that the initial velocity of the reaction (V0), maximal amount of the reaction product (Pmax) and characteristic time of the reaction (T) comprised 1.8 +/- 0.4 and 1.6 +/- 0.2 mmole Pi/min per 1 mg protein, 7.5 +/- 1.3 and 1.4 +/- 0.2 mmole Pi/mg protein and 4.1 +/- 0.7 min and 1.1 +/- 0.1 for Ca2+-ATPases from plasma and endoplasmic reticulum membranes, correspondingly. High- and low-affinity sites of ATP and Ca2+-binding in Ca2+ ATPases from plasma and endoplasmic reticulum membranes were identified. Negative cooperation in ATP binding to Ca2+-ATPase from plasma membrane and a positive cooperation for Ca2+-ATPase from endoplasmic reticulum has been found. Ca2+ binding to low-affinity sites of both Ca2+-ATPases showed no cooperation, while Ca2+ binding to high-affinity sites showed the positive cooperation. Using the Hill's coordinates we have found the values of the Mg2+ Michaelis constant (K(Mg)) which yielded 3.89 x 10(-5) and 3.80 x 10(-5) mole/l for Ca2+-ATPases from plasma and endoplasmic reticulum membranes, correspondingly. It is supposed that obtained data are important for further studies of molecular and membrane mechanisms involved in the regulation of intracellular calcium signalling and secretion by salivary acinar cells. PMID- 16350745 TI - [Calcium homeostasis during thymocyte apoptosis. I. Increase in cytosolic Ca2+ concentration at the early stage of apoptosis induced by hydrogen peroxide]. AB - The concentration of free cytosolic Ca2+ ([Ca2+]i), 45Ca2+ entry and the level of reduced glutathione (GSH) after x-irradiation in a dose of 4.5 Gy or 0.1 mM H2O2 treatment were investigated in isolated rat thymocytes during the period preceding electrophoretically detected DNA intranucleosomal fragmentation. Using fura-2 it was shown that the level of [Ca2+]i in X-irradiated thymocytes was not changed as compared with the control, while the GSH content was increased. The gradual increase in [Ca2+]i along with GSH level falling was detected in the H2O2 treated cells. 45Ca2+ entry in the cells exposed to apoptogenic stimuli was not enhanced. After addition of H2O2 to the cells previously treated with thapsigargin further [Ca2+]i increase in both normal and nominally calcium-free medium was detected. Cyclosporine A inhibited Ca2+-mobilizing effect of H2O2, but did not prevent it completely. The role of intracellular calcium depots in calcium homeostasis disturbance during oxidative stress and apoptosis is discussed. PMID- 16350744 TI - [Influence of Mg ions and spermine on ATP-dependent Ca2+ transport in myometrial intracellular structures. II. Comparative study of spermine, Mg ions and cyclosporin A effects on Ca2+ transport in mitochondria]. AB - In experiments carried out with the use of the radioactive label (45Ca2+) on suspension of the rat uterus myocytes processed by digitonin solution (0.1 mg/ml), influence of spermine and cyclosporin A on Mg2+, ATP-dependent Ca2+ transport in mitochondria at different Mg2+ concentration were investigated. Ca2+ accumulation in mitochondria was tested as such which was not sensitive to thapsigargin (100 nM) and was blocked by ruthenium red (10 microM). It has been shown, that spermine (1 mM) stimulates Mg2+, ATP-dependent Ca2+ accumulation in mitochondria irrespective of Mg2+ concentration (3 or 7 mM) in the incubation medium. At the same time cyclosporin A (5 microM) effects on Ca2+ accumulation in mitochondria depend on Mg2+ concentration in the incubation medium: at 3 mM Mg2+ the stimulating effect was observed, and at 7 mM Mg2+ - the inhibitory one. In conditions which led to the increase of nonspecific mitochondrial permeability and, accordingly, to dissipation of electrochemical potential (it was reached by 5 min. preincubation of myocytes suspension in the medium that contained 10 microM Ca2+, 2 mM phosphate and 3 or 7 mM Mg2+, but not ATP) significant inhibition of Mg2+, ATP-dependent Ca2+ accumulation in mitochondria was observed. The inhibition to the greater degree was observed when medium ATP and Mg2+ were absent simultaneously in the preincubation. Thus the quality of spermine effects on Ca2+ accumulation was kept: stimulation in the presence both of 3 mM and 7 mM Mg2+. Ca2+ accumulation did not reach the control level when 3 mM Mg2+ and 1 mM spermine was present and ATP absent in the preincubation medium. However, in the presence of 7 mM Mg2+ and 1 mM spermine practically full restoration (up to a control level) of Ca2+ accumulation was observed. At the same time with other things being equal such restoration was not observed at simultaneous absence of ATP and Mg2+ in the preincubation medium. The quality of cyclosporin A effects on Ca2+ accumulation in mitochondria was also kept: stimulation - in the presence of 3 mM Mg2+, inhibition - in the presence of 7 mM Mg2+ in the preincubation medium. And, at last, in the presence of cyclosporin A irrespective of the fact which preincubation medium was used, Ca2+ accumulation level practically did not depend on Mg2+ concentration. PMID- 16350746 TI - [The factors of antioxidant defence in the blood and liver of pheasants during ontogenesis]. AB - The changes in activity of antioxidant enzymes of LPO products and lipophilic bioantioxidants of pheasants during the period from 1 day up to 60-day age was investigated for the first time. It is shown, that insufficiently generated enzymatic antioxidant systems is a cause and result of high intensity of processes of free-radical LPO in 1-day pheasants tissues. The compensated increase of antioxidant enzymes (SOD, CAT) activity during the early period of postnatal ontogenesis does not prevent from the fast exhaustion of a pool of the main lipophilic bioantioxidants that in its turn provokes the inhibition of antioxidant system enzymes reactions. This results in the decrease of adaptability of wild birds in conditions of artificial breeding. PMID- 16350747 TI - [Effects of spermine and putrescine on the binding of the transcription factor NF kappaB to specific DNA sequences]. AB - Using fluorescence resonance energy transfer technique, effects of polyamines (spermine and putrescine) on the binding of p50 subunit of the nuclear transcription factor NF-kappaB to specific DNA sequences were studied in model experiments. It is demonstrated that polyamines in the concentration range from 1 to 500 mcM facilitate the binding of NF-kappB to specific DNA sequences. The data currently obtained agree with our previous predictions made by the computer modelling techniques. Our results presented in this work allow us to suggest that polyamines (excessive synthesis of which proceeds in tumor growth) may participate in the regulation of transcription of several oncogenes by the factor NF-kappaB. PMID- 16350748 TI - [Peculiarities of pH influence on aldehyde reductase activity in the liver of rats from different age groups subject to immobilization stress]. AB - In order to develop the notions about the causes of age-dependent changes in the organism resistance to the influence of damaging stress factors the study of aldehyde reductase activity manifestations was carried out in the liver of 1.5-, 2-, 6-, 12-, 24-months-old rats subject to immobilization stress with different values of pH of reactive mixture. It has been shown that the immobilization stress is accompanied by the shift of the enzyme pH-optimum to the acid side. This change manifestation in 1.5-months-old and 24-months-old animals has been less significant as compared with that in the rats of other age groups. PMID- 16350749 TI - [Redox-state of NAD pairs and activity of lactatedehydrogenase and NADase in guinea pig tissues at different stages of development of experimental allergic encephalomyelitis]. AB - Changes of NAD content, redox-state, enzyme activity in the brain and liver tissues of Guinea pigs at different stages of development of experimental allergic encephalomyelitis (EAE) were investigated using the model of multiple sclerosis. It was shown, that the most legible changes of the investigated parameters occur on the preclinical stage and the stage of initial neurological symptoms. The increase of the brain NAD level and reduction properties of NAD+/NADH pairs reduction properties against a background of inhibition of lactatedehydrogenase activity was observed in the early terms of EAE development (7-15 day). The liver lactatedehydrogenase activity is increased at an initial stage. NAD-ase activity is increased in the medium term (18-21 day) that correlates with changes of this enzyme activity in the blood serum. In the term of strongly expressed neurological signs (26-33 day) the sharp drop of NAD content in the brain and liver is observed. The role of the obtained results at different stages of EAE development is discussed. PMID- 16350750 TI - [Age-dependent cadmium accumulation in organs of intoxicated rats and changes of indices of acid-base balance of blood in different conditions of antioxidant defence]. AB - Age peculiarities of cadmium cumulation in the organs of rats under different conditions of antioxidants administration. Considerable hepato- and nefrotoxic effect of the given xenobiotic under different conditions of its supply into the organism was revealed. The strongly pronounced differences between the age distribution of cadmium in the organisms have been marked. It is proved that the most antioxidantal activity is shown by sodium selenite when administered into the organism before poisoning with cadmium. It is stated that the indices showing the acid-alkaline state are subject to the greatest changes in the bodies of young animals. In this case they have more expressed toxic effects under the influence of ions of heavy metals as well as the receptivity to the positive influence of antioxidants. PMID- 16350751 TI - [Effect of iron ions on the antioxidant enzyme activities in yeast Saccharomyces cerevisiae]. AB - A complex of physiological and biochemical indices has been compared in wild and isogenic catalase-deficient strains of Saccharomyces cerevisiae grown on the media with different iron ion concentrations is 2 times higher in cytosolic catalase deficient yeast. Superoxide dismutase activity grown in the medium with 500 microM of ferrous sulphate. Under such conditions, peroxisomal catalase deficient yeast had a 2-fold decreased activity of superoxide dismutase. There is a significant difference between TBA-reactive substances content of the wild and cytosolic catalase deficient strain. It has been suggested that the repletion of iron ions in the growth medium leads to the formation of lipid oxidation products. Catalase prevents TBA-reactive substances formation in the given conditions and plays a protective role. PMID- 16350752 TI - [Na+, K+-ATPase activity and serotonin transport in the rat brain synaptosomes fractions after acute 1,2-dichloroethane intoxication and nicotinamide administration]. AB - Alterations of Na+,K+-ATPase activity and serotoninergic system functioning were investigated in brain synaptosomes fractions of rats under experimental acute 1,2 dichloroethane (DChE) intoxication. It was shown that Na+,K+-ATPase activity was markedly increased (by 41,8%) in a period of 24 h after DChE intoxication and decreased (by 27%) after 48 h intoxication. The level of [2-14C]-serotonin uptake by synaptosomes was progressively diminished after 24 and 48 h after DChE injection whereas the activity of monoamine uptake proved to be unchanged. Nicotinamide (200 mg/kg of body weight) was administered to rats subjected to DChE 1, 24 and 36 h after poisoning. The treatment of rats with nicotinamide resulted in some normalization of brain synaptosomal Na+, K+-ATPase activity and serotonin uptake controlled at 48 h after DChE intoxication. PMID- 16350753 TI - [Activation of reparative processes by phospholipid preparation in the affected organs and tissues in neonathal enteropathology of calves]. AB - The peculiarities of dynamics of quantitative changes of some classes of lipid and phospholipid spectra of blood plasma of calves recovered after dyspepsia were studied. Obtained reliable changes of the blood plasma lipidogrammas testify to development of dyslipidemia. It is characterized by hypercholesterolemia and hypertriacylglycerolemia of recovered 30 days old calves 3 weeks after diseases symptoms past. These changes give evidence concerning deficiency of phosphatides choline fraction - main structural components of cell membranes. It was established that changes of lipid and phospholipid spectra of blood plasma caused by enteropathology can be corrected by the inclusion of reparative therapy preparations to dyspepsia treatment plan in particular--experimental phospholipid containing a drug, which is prepared on the basis of milk phospholipids--its natural source for newborn calves. PMID- 16350754 TI - [Anticonvulsive activity of antiepileptic drugs and quantum-chemical modelling of their interaction with GABA(A) receptor]. AB - The results of experimental analysis of the clinical activity of the antiepileptic drugs' (Phenobarbital, Carbamazepine, Valproic acid, Lamotrigine, Topiramate, Felbamate) widely used in clinic, that was carried out using the standard convulsion test with bicuculline in vivo were compared with characteristics of these drugs' interaction with the key aminoacids of GABA(A) receptor calculated by quantum chemical method (program HyperChem7, semi empirical method AM1 technique). The correlation between the activity of the drugs in the experiment in vivo and energy of system's interaction of the drugs with aminoacid residue Thr201-Thr202-Gly203- Ala204-Tyr205-Pro206 was found out. PMID- 16350755 TI - [ATPase and phosphatase activity of drone brood]. AB - Most researches on insect enzymes concern carbohydrate and nitrogenous exchange. Data on ATPase activity for larval material of drone brood are absent in the available literature. The drone brood is one of the least investigated apiproducts. Allowing for the important role of ATPase in the vital functions of the insect cells our work was aimed at the study of ATPase of the drone blood activity and that of alkaline and acid phosphatases. When studying liophylised preparations of the drone brood homogenate we have found out high activity of Mg2+, Na+, K+-, Ca2+- and Mg2+-ATPase and of alkaline and acid phosphatase, that is the possible explanation of the high-intensity power and plastic processes proceeding during growth and development of larvae. PMID- 16350756 TI - [Evaluation of intramolecular interaction between complementary domains, connected with a flexible chain]. AB - A method for the evaluation of the effective binding parameters for the interaction between two complementary domains connected with a flexible chain is suggested. The calculations are based on the assumption that the chain is absolutely flexible and does not hinder free relative diffusion of the domains in the solution but does not allow the domains to move away from each other further than the length of the chain. Then, if the distance between the connected domains and the affinity of the interaction between disconnected domains are known, the suggested method allows calculation so-called "local concentration" of the domains relative to each other. On this basis, it is possible to estimate the upper limit to the fractional content of domains in complex, which, when constrained by the linking polypeptide chain, may be much higher than implied by the absolute concentrations of the domains. PMID- 16350757 TI - [New approach to an increase of specificity in determination of antibodies to lipopolysaccharides of Gram-negative bacteria by solid-phase enzyme-linked immunosorbent assay]. AB - A simple and effective method is proposed to increase the sorption of lipopolysaccharides (LPS) of Gram-negative bacteria on the polystyrene surfaces, in which there is no need of preliminary modification of the surface of the solid phase with poly-L-lysine. The method consists in using 0.05 M carbonate buffer (pH 9.6) with 70% of ammonium sulfate at the stage of LPS immobilization. As a result, LPS absorption increases essentially, and the level of non-specific reactions remains low. This makes the usage of additional blocking reagents unnecessary and allows increasing sensitivity and specificity in determination of anti-LPS-antibodies by the solid-phase enzyme-linked immunosorbent assay. PMID- 16350758 TI - [Quantification of D-dimer and soluble fibrin in blood plasma of people with ischemic heart disease and hypertension]. AB - The method of D-dimer quantification in the human blood plasma has been developed using monoclonal antibodies 111-3b and II-4d. The method has been verified on the blood plasma of the patients with ischemic heart disease with and without stenocardia and with hypertension. The results showed that at ischemic heart disease with and without stenocardia and at hypertension the quantities of D dimer in the blood plasma were generally less than the highest normal level 500 ng/ml (64.3%, 76.2% and 95%, correspondingly). The semiquantitative measurements of soluble fibrin levels in blood plasmas of the patients with ischemic heart disease and hypertension have been performed. It has been shown that the quantity of soluble fibrin at these diseases range greatly from < 0.03 mg/ml to 0.15 mg/ml. There was no correlation between the quantities of D-dimer and soluble fibrin in blood plasmas of the patients. Electrophoresis in PAAG with SDS showed that the soluble fibrin at these diseases had the mo- lecular mass of the fibrin (ogen). Thus the soluble fibrin in blood plasmas analysed consisted mainly of fibrin desAA oligomers (may be with fibrinogen incorporation) which are not stabilized by the factor XIIIa. PMID- 16350759 TI - [Kinetic regularities of the proceeding and possible reaction mechanism of Mg2+ dependent enzymatic hydrolysis of ATP in the fraction of plasmatic membranes of the smooth muscle]. AB - Kinetic regularities of the reaction of Ca2+-independent Mg2+-dependent enzymatic hydrolysis of ATP catalyzed by the so-called "basal" Mg2+-ATPase localized in the plasmatic membrane of the uterus smooth-muscle cells have been studied using the methods of kinetic analysis performed under the equilibrium conditions. The analysis was based on the study of the concentration dependence of initial velocity of nucleoside triphosphate hydrolysis in EGTA-containing medium under the change of general concentrations of ATP [ATP]o and Mg2+[Mg2+]o in conditions of their equimolar ratio ([ATP]o/ [Mg2+]o)= 1; here the ratio between the concentrations of free reagents ([ATP4-]o/[Mg2+]o) was equal to 1.25. The obtained concentration dependence was interpreted in terms of two practically possible alternative mechanisms of Mg2+-dependent ATP-hydrolase enzymatic reaction. Mechanism I. Two separate independent centres of Mg ions and ATP binding by the enzymatic protein are supposed to exist, while Mg2+-dependent ATP hydrolase enzymatic reaction proceeds independent of the equilibrium reaction of Mg ions chelatization of muscleside triphosphate. Mechanism II. The existence of the only centre of the chelate complex Mg2+ATP2- binding is postulated on the enzymatic protein; this process is also realized independent of the binding of Mg2+ and ATP-hydralase reaction catalized by it. PMID- 16350760 TI - [From bioinformatics to systems biology: account of the 12th international conference on intelligent systems in molecular biology]. AB - The paper reviews the 12th International Conference on Intelligent Systems for Molecular Biology/Third European Conference on Computational Biology 2004 that was held in Glasgow, UK, during July 31-August 4. A number of talks, papers and software demos from the conference in bioinformatics, genomics, proteomics, transcriptomics and systems biology are described. Recent applications of liquid chromatography - tandem mass spectrometry, comparative genomics and DNA microarrays are given along with the discussion of bioinformatics curricular in higher education. PMID- 16350761 TI - The case for alcohol research as a focus of study by nurse researchers. AB - Sixty percent of the U.S. population drinks alcohol. Although numerous investigators have shown that low-volume alcohol intake has positive influences on cardiovascular health, bone density, and cognition, there is a fine line between positive, neutral, and negative ramifications of alcohol consumption on health. Alcohol accounts for 7% of the global burden of disease and injury from all causes and for 10% to 11% of all illnesses and death each year worldwide. So alcohol use is a two-edged sword. Psychiatric nurses have a long history of involvement with alcoholic patients, and alcohol users, misusers, and abusers comprise a significant percentage of the patient load in every specialty and subspecialty of nursing. Yet nursing education has neglected this important area of content in general nursing curricula, staff development has not trained mainstream nurses to routinely assess for alcohol problems among hospital patients, and primary care providers have failed to do case finding. During the past 25 years, the federal government has funded curricular and faculty development programs to bring alcohol information into the core of health provider training. The process has been halting and minimally successful at best. One ramification of the inattention to alcohol-related education is the dearth of nurse-scientists engaged in programs of research in the field of alcohol abuse. A federally funded faculty development program in the 1990s focused attention on this issue, and a small cadre of nurses were trained to do related research. Several of the authors in this volume are members of that group. A brief overview of the focus of each of the remaining chapters in this volume is presented. A rationale is provided for the importance of this area of research for nursing knowledge and quality patient care in essentially all areas of nursing practice. Finally, several burning research questions are posed that would most appropriately be answered through nursing research. PMID- 16350762 TI - Measurement of alcohol consumption: issues and challenges. AB - In both the clinical and research settings, nurses assess patterns of alcohol consumption to screen for risk of adverse events or to determine the health consequences of drinking. The purposes of this critical review are to explore issues and controversies surrounding the measurement of alcohol consumption and to critique the existing literature relevant to the research and clinical arenas. An electronic literature search was completed to identify research articles addressing human studies from 1995 through 2004 related to alcohol consumption. Key words included alcohol drinking (subheadings blood, metabolism, psychology, and urine), standard drink, problem drinking, heavy drinking, and ethanol analysis (subheadings blood, urine, and chemistry). The results were in two primary content areas: self-reported alcohol consumption and assessment of consumption by using biological markers. Self-reported alcohol consumption can be quantified in a variety of ways, such as ounces of ethanol per day, standard drinks per day, drinking occurrences per month, heavy drinking occasions per month, and frequency of perceived drunkenness. The choice of measure depends on setting (clinical vs. research), the role of the variable under study, the capabilities and demographics of the study population, the study design, and the resources available to collect alcohol consumption data. A variety of biologic instruments are used to assess alcohol consumption, each with sensitivities and specificities that vary by age, gender, and possibly by ethnicity/race. Previous work has focused on the white, male, alcohol-dependent population and non-alcohol dependent male controls. Some urgency exists to expand the biometrics of alcohol use to minority and older populations as well as to women across the life span. PMID- 16350763 TI - Moderate drinking and cardiovascular disease. AB - The adverse consequences of heavy alcohol use are well known. However, recent media reports of a possible cardiovascular benefit associated with moderate drinking have revived public interest in the use of alcohol for "medicinal purposes." Knowledge development regarding guidelines for moderate alcohol use has lagged behind public interest in the possible health benefits of moderate alcohol use. At this time, evidence-based primary health promotion interventions related to the risks and benefits of moderate alcohol use are lacking in the health care literature. This chapter reviews 22 reports describing the relationship between moderate drinking and cardiovascular disease. The reports are classified by the level of evidence and critiqued on seven aspects of method. Conclusions related to the strength of the evidence that moderate drinking is a useful primary health promotion intervention are presented. PMID- 16350764 TI - Alcohol consumption during pregnancy. AB - Alcohol is a potent teratogen in humans, and prenatal alcohol exposure is a leading preventable cause of birth defects and developmental disabilities. The term fetal alcohol syndrome (FAS) refers to a pattern of birth defects found in children of mothers who drank during pregnancy. FAS has four criteria: maternal drinking during pregnancy, a characteristic pattern of facial abnormalities, growth retardation, and brain damage (often manifested by intellectual difficulties or behavioral problems). As surveillance and research have progressed, it has become clear that FAS is but a rare example of a wide array of defects that can occur from exposure to alcohol in utero. At least 1 in 10 women will continue to consume alcohol during pregnancy, putting their fetuses at risk for the effects of alcohol exposure. Nurses are in a key position to provide care and conduct research that will contribute to the prevention of the adverse effects of prenatal alcohol exposure during the preconception and perinatal periods, as well as deal with the negative outcomes of exposure in the developing infant. Many areas have yet to be evaluated. Screening tools and interventions have been developed and tested, mostly in majority cultures. Culturally sensitive instruments must be generated and validated for high-risk groups such as Native Americans. Fetal alcohol biomarkers and genetic research are new and need considerably more work. Effective "no drinking during pregnancy" campaigns for high-risk groups must be created and tested. Nurses are well placed to conduct research that will describe the effects at social, behavioral, and biological levels; develop middle-range theories targeted at preventing the drinking behavior and optimizing care of affected children after birth; and generate and test effective interventions that enhance prevention strategies in the 21st century. PMID- 16350765 TI - Alcohol, children, and adolescents. AB - Alcohol use in children and adolescents continues to be a major health concern. There is a rich literature on correlates and antecedents of alcohol use in children and adolescents, and concerted efforts have been made to develop, implement, and evaluate intervention strategies. This chapter provides a review of the intervention studies to prevent alcohol use in these groups. The interventions are categorized by their primary focus: school, family, and community. The studies were limited to those with either an experimental or a quasi-experimental design and published results. PMID- 16350766 TI - College students' alcohol use: a critical review. AB - This integrative review of college students' alcohol use covers research papers as well as review and theoretical papers published between 1990 and 2004. To conduct this review, abstracts were identified by searching Medline (PubMed), Ingenta, ERIC, PsycInfo, and Health Reference Center Academic using the following words: alcohol and college drinking, binge drinking, college students and undergraduates and the years 1990 to 2004. From an initial list of over 400 abstracts, 203 papers were identified and considered for this review. A developmental perspective of college drinking was assumed, and the chapter is organized within five domains: biology, identity, cognition, affiliation, and achievement. In addition, research pertaining to the harmful consequences of college drinking and the assessment of risky drinking is reviewed and discussed. The chapter concludes with the identification of gaps in knowledge and implications for future research. PMID- 16350767 TI - Alcohol misuse, abuse, and addiction in young and middle adulthood. AB - This chapter reviews research on alcohol misuse, abuse, and addiction in young and middle adulthood. Young adulthood is defined as ages 21 to 35 and middle adulthood as ages 36 to 65. The authors searched the Cumulative Index of Nursing and Allied Health Literature (CINAHL) and other databases for the years 1992 through 2004 using the terms alcohol abuse, alcoholism, and alcohol dependence, then hand-searched for the inclusion of age as a variable. The search was limited to research and English-language publications. Unpublished dissertation studies were excluded, as were topics that were reviewed in other chapters of this volume. When possible, articles for review were limited to alcohol only, as compared with other drugs of abuse. Research articles were selected for review if they contained the variables alcohol misuse, abuse, or alcohol addiction or dependence and age. The review included both nurse- and non-nurse-investigators and was comprised of 50 studies. The results are in four content areas across the global area of alcohol misuse, abuse, and dependence in young and middle adulthood: incidence and prevalence, developmental changes, the work setting, and the family setting. Few articles used a developmental framework, although in some studies, the framework was implicit. Nurse-investigators were more likely to produce qualitative studies, although the studies varied dramatically in size of sample, research design, and variables. Overall, there was greater breadth than depth. The use of adult developmental theory was limited despite its potential explanatory potential in this field. There is a need for more nurse-researchers to explore adult developmental theory and pay increased attention to age and developmental stage as explanatory variables. PMID- 16350768 TI - Alcohol use, misuse, abuse, and dependence in later adulthood. AB - Considerable research has focused on alcohol problems in older adults, but the clinical utilization of this knowledge has lagged at least 3 decades behind the scientific developments. This unfortunate situation takes on added significance as the "baby boomer" generation ages because more of them drink more often in larger quantities than previous generations. This chapter focuses more on the ramifications of use, misuse, and abuse than on chronic dependence because the prevalence in the former categories far outweighs the latter. Older alcohol misusers and abusers are at excess risk for myriad physical problems and premature death because alcohol interacts with the natural aging process in negative ways to increase risks for injuries, hypertension, cardiac dysrhythmic events, cancers, gastrointestinal problems, neurocognitive deficits, bone loss, and emotional challenges, most notably depression. Low volume and less than daily alcohol consumption appear to be protective against blood clots in the coronary and brain vessels, bone loss and falls, and cognitive decline compared with current abstainers. At higher levels, alcohol has the opposite effect. Research findings strongly support positive outcomes of case finding, referral, and treatment of older adults who are misusing or abusing alcohol. However, there is ample evidence that health care providers across the spectrum of primary, acute, and long-term care ignore the signs and symptoms of alcohol misuse and abuse in their older patients and treat symptoms and sequelae of the abuse rather than confronting the abuse itself. Recommendations for changes in practice are made together with ideas for additional research in several areas where the current state of knowledge is inadequate, conflicting, or based on narrowly homogeneous samples. PMID- 16350769 TI - Alcohol and risky behaviors. AB - The purpose of this chapter is to review and critique the literature on risky drinking, driving, and sexual behaviors. To complete this review, electronic searches using databases from the disciplines of nursing, medicine, and psychology were used with keywords alcohol and risky behavior, risky drinking, risky driving, risky sex, and sexual aggression, as well as other relevant terms. The basic tenets of contemporary theoretical models of risky behaviors are used as a framework for reviewing the literature. Most relevant to the discussion are the relationships among the behaviors, risk and protective factors, and major unresolved theoretical and methodological issues. In the literature, sensation seeking was differentially associated with risky drinking, driving, and sex, but causal assertions are premature. Important conceptual and physiological issues are clarified. First, unconventionality contributes to risky drinking, risky driving, and, among adolescents, risky sex. Second, the pharmacologic effects of alcohol on cognitive processing contribute to risky sex, but only among persons who feel conflicted about risky sex (e.g., condom use). This perception may be particularly true for men who have a belief that alcohol will enhance sex. Third, sexual aggression appears to stem from a variety of factors, including the pharmacologic effects of alcohol on aggression and stereotypes about drinking women. Exploration of risk and protective factors adds breadth and depth to the discussion of risk taking. Risk factors include (1) high tolerance for deviance, (2) unconventional attitudes and behaviors such as early alcohol use and precocious sex, (3) peer norms for deviance, (4) high sensation seeking, and, to a lesser extent, (5) disturbed risk perception and positive beliefs about alcohol. Protective factors appear to mitigate risk and include (1) conventional attitudes and behaviors and (2) having peers that model conventional attitudes and behaviors. Although empirical evidence suggests that risky behaviors tend to covary, most intervention trials to date have focused on single behaviors, and often are based on clinical information rather than existing theoretical and empirical knowledge. PMID- 16350770 TI - Alcohol brief interventions. AB - A large proportion of Americans report binge or heavy drinking. The human and economic costs of alcohol misuse are extensive, with hundreds of thousands of lives lost or disrupted and billions of dollars spent due to impaired productivity, crime, and adverse health consequences. In an effort to reduce costs such as these, scientists and clinicians have developed brief interventions, characterized by their low intensity and short (5-60 minutes) duration, as well as by their intent to provide early intervention before drinkers develop alcohol abuse or dependence. The purpose of this review, therefore, is to analyze research studies related to brief intervention and critically analyze and critique their findings. In addition, both prospective randomized controlled trials and meta-analyses will be used to discuss the implications for clinical practice and make recommendations for future research. PMID- 16350771 TI - Promises, promises: promise yourself a healthier lifestyle. PMID- 16350772 TI - Your child's health: allergies. PMID- 16350773 TI - Yoga & Tai Chi: stress management and low impact fitness from the East. PMID- 16350774 TI - How low should your carbs go? PMID- 16350775 TI - Elderly drivers: when to hang up the keys. PMID- 16350776 TI - Need help understanding Medicare or Medicaid? PMID- 16350777 TI - The doctors will e-mail you now. PMID- 16350778 TI - May we see your records? PMID- 16350779 TI - The republican who thinks big on health care. PMID- 16350780 TI - A transplant first. PMID- 16350781 TI - A better CPR. PMID- 16350782 TI - [A study on peak compression effect in ion-exchange capillary electrochromatography with relaxation theory]. AB - Due to the electric characteristics of the solute transport in capillary electrochromatography (CEC), some special phenomena might occur. An example is the peak compression effect in ion-exchange capillary electrochromatography, which cannot be explained with common chromatography theory. According to the basic model of relaxation theory, it is considered that there are "co-" and "counter-" solute flows in mobile phase and in stationary phase, the expressions of the first moment and the second center moment of profile, and column efficiency are derived by the method of Laplace transform. The relations of electrophoresis velocity, electroosmosis velocity and electric dispersion velocity are also investigated with these expressions. The results show that this new model offers a better explanation for peak compression effect on the migration behaviour of charged solute in CEC. The retention time will be shorter by peak compression effect, which is similar to a solvent gradients process. The peak compression effect is not steady, and doesn't appear until the conditions are fitted in some special situation. PMID- 16350783 TI - [Correlation between octanol-air partition coefficients and retention parameters of polychlorinated biphenyls on gas chromatographic columns]. AB - Octanol-air partition coefficient (Koa) is a key descriptor for describing the partitioning of organic pollutants, especially persistent organic pollutants (POPs), between air and environmental organic phases. Thus it is necessary to determine or predict the Koa values of these compounds. A novel method was introduced to describe the quantitive relationship between the octanol-air partition coefficients and gas chromatographic (GC) retention parameters A, B values of polychlorinated diphenyls (PCBs) which are typical persistent organic pollutants. By the correlation analysis between GC retention parameters A, B values of PCBs on DB-1, DB-5 and DB-1701 columns, and their octanol-air partition coefficients, a good linear relationship was found. The stepwise multiple linear regression was used to derive binary equations with the correlation coefficients greater than 0.99, and the standard deviations are less than 0.11. In addition, based on the 153 experimental A, B values and 56 predicted A, B values, the Koa values of the remaining 190 PCBs were predicted. PMID- 16350784 TI - [Quantitative relationship between molecular structure of polychlorinated biphenyls (PCBs) and enthalpy change (deltaH), entropy change (deltaS') in chromatographic process]. AB - The relationship between the rule of chromatographic retention value and molecular structure is an important part in the research of chromatographic thermodynamics. The topological index structural parameter JG and the topological index adjoining parameter LJ are put forward. Parameter J(G) describes the correlation of quantity and position of chlorine atoms in polychlorinated biphenyl (PCB) molecules. Parameter L(J) describes the ortho-position correlation of chlorine atoms in PCB molecules. The relational expression between the PCB molecular structures and their enthalpy change (deltaH), entropy change (deltaS') in chromatographic process was discovered. The values of enthalpy change and entropy change for about 140 kinds of polychlorinated biphenyls in chromatographic process on three stationary phases, DB-1, DB-5 and DB-1701, were determined. In comparison with deltaH and deltaS' of the experimental data those calculated from the relational expression had the average relative deviations for deltaH and deltaS' are 0.56% -0.97% and 0.55% - 1.06%, respectively. PMID- 16350785 TI - [Applications and progresses of expert system on chromatography]. AB - The expert system on chromatography has achieved great advancement in the past two decades, and is playing a more and more important role in solving analytical problems of complex samples. Research results of expert system on chromatography in authors' group are reviewed with 64 references. A brief introduction of the expert system on chromatography is presented. Applications of the expert system on chromatography are summarized in the fields of petrochemical online analysis, environmental air sample analysis, tumor diagnosis and traditional Chinese medicine analysis. The review followed the scientific foot steps in the authors' group, starting from the development of the expert system on gas chromatography, to the selection of multi-column systems in online industrial gas chromatographs in petrochemical plants, and to the employment of the new techniques in gas chromatography, liquid chromatography and capillary electrophoresis to solve the practical analytical problems in the nation's scientific and economic development. PMID- 16350786 TI - [Laboratory on a microfluidic chip]. AB - The recent achievements of microfluidic chip and its applications, based on the works mainly carried out in the authors' lab are reviewed. The chip fabrication capabilities have been extended into design and fabricate chips with higher degree of complexity in different materials, such as quartz, glass, polymethyl methacrylate (PMMA), and polydimethyl siloxane (PDMS). A set of methods for surface modification of micro-channels on such materials have been developed, which results in better reproducibility and higher efficiency in protein and peptide analysis. The use of novel materials for chip fabrication is also under investigation. A series of microfluidic workstations with integrated chip manipulation as well as laser induced fluorescence (LIF), ultraviolet (UV), electrochemical and chemiluminescence detection modules have been developed to attain the abilities of complex microfluidic control and data acquisition schemes. A single cell/single molecule imagining system was built up for dynamic analysis of molecular or cellular events too. Based on the work mentioned above, different functional units, such as membrane, monolithic, isotachophoresis (ITP) etc were set up and integrated. Glycoform separation of turkey ovalbumin in a lectin monolithic column and an electrophoresis channel was performed on an integrated microchip. And a novel technique has been developed that allows for the coupling of ITP and non-gel sieving electrophoresis for protein analysis in a single microchip and resulting in - 50 fold increase of the sensitivity in comparison with the use of gel electrophoresis only. A single molecule detection (SMD) based technique was developed for simultaneously measuring both bulk flow and near-wall flow velocity in the microchannels. And more recently, an SMD based technology was developed for observing molecular interactions at single molecule level. An ultra-rapid microchip electrophoresis method was established for simultaneous determination intracellular reactive oxygen species (ROS) and reduced glutathione (GSH) related to apoptosis and oxidative stress. In an effort to develop a novel microfluidic based drug screening platform, systematic studies on the interaction between granulocyte colony-stimulating factor (G-CSF) and sulfated oligosaccharides were carried out at both molecular and cellular levels. Doxorubicin induced apoptosis of human hepatocellular carcinoma (HepG2) was studied using the integrated microfluidic device with concentration generator. In the application phase, severe acute respiratory syndrome (SARS) diagnosis based on reverse transcription-polymerase chain reaction (RT-PCR) and microfluidic chip electrophoresis (MCE) with 18 cases, methylation analysis of the P16 gene in 159 samples of patients and references for cancer diagnosis and polymorphism analysis of angiotenigen gene in 226 patients and references with essential hypertension are described. Forty-three up to date references are PMID- 16350787 TI - [Advances in packing capillary electrochromatographic columns]. AB - The columns in capillary electrochromatography can be classified into three classes: open tubular, packed and monolithic columns. The monolithic columns can be divided into three categories: organic polymer-based monolithic columns made from the polymerization of acrylamide, styrene, acrylate or methacrylate monomers, silica-based monolithic columns generally prepared by using sol-gel technology, and packed particulate-based monolithic columns. Monolithic columns are receiving quite remarkable attention and developing rapidly with a focus on monolithic stationary phases prepared from synthetic polymers. The preparation methods for various types of capillary electrochromatographic columns and their advantages and disadvantages are reviewed according to 100 research articles. In particular, recent advances in the preparation methods of monolithic columns and their applications are discussed in details. PMID- 16350788 TI - [Screening of antineoplastic components in Radix et Rhizoma Rhei using chromatographic fingerprints]. AB - A new strategy for screening of antineoplastic components in the traditional Chinese medicine of Radix et Rhizoma Rhei has been developed using chromatographic fingerprints before and after metabolism by high performance liquid chromatography-mass spectrometry. The metabolizing method was established based on the in vitro metabolism by Sprague-Dawley (SD) rat liver homogenate. By means of the metabolism methods in vitro, the antineoplastic activity of the extracts, metabolites and components of Radix et Rhizoma Rhei were determined by microculture tetrazolium (MTT) assays in vitro. It was observed that the inhibition rate of the crude extract for HeLa cell was increased from 26.7% to 36.2% after 60 min of metabolism. The changes of activities resulted from the changes of components' structures and the bioactive components were discovered simultaneously in view of metabolism by inhibiting rate assay for the components in Radix et Rhizoma Rhei. It is concluded that the antineoplastic activity of the crude extract from Radix et Rhizoma Rhei was increased after in vitro metabolism because the antineoplastic activity of aloe-emodin, the metabolite of chrysophanol, is higher than its parent compounds. PMID- 16350789 TI - [Separation and identification of isoflavonoids in Pueraria lobata extracts and its preparations by reversed-phase capillary liquid chromatography coupled with electrospray ionization quadrupole time of flight mass spectrometry]. AB - Isoflavonoids in Pueraria lobata extract and its preparations were separated and identified by reversed-phase capillary liquid chromatography (RP-CapLC) coupled with photodiode array (PDA) detector and negative electrospray ionization quadrupole time of flight (Q-TOF) mass spectrometry. The separation was performed on a 150 mm x 0. 32 mm i. d. , 5 microm C18 capillary column, using mobile phase of 0.1% aqueous trifluoroacetic acid solution and acetonitrile containing 0.1% trifluoroacetic acid under gradient elution. The product ion spectra of the deprotonated ions allowed for the identification of puerarin, daidzin and daidzein in the sample. Puerarin was found to be the most abundant component in the extract (about 13%, mass fraction) and its preparations (19.28 - 24.34 mg per tablet). The structures of trace amount of unknown isoflavonoids were deduced based on the spectra of known compounds. They were proposed to be 3' methoxypuerarin and 3'-methoxydaidzin. PMID- 16350790 TI - [Simultaneous determination of five groups of components in qingkailing injection by high performance liquid chromatography with photo diode array detector and evaporative light scattering detector]. AB - A method was established for the simultaneous quantification of nine components of five different structural types in Qingkailing injection. High performance liquid chromatography coupled with a photo diode array detector and an evaporative light scattering detector (HPLC-DAD-ELSD) was employed in the determination. Four monitoring wavelengths of 240, 254, 280 and 330 nm were set to determine nucleosides (uridine and adenosine), iridoid glucoside (geniposide), flavone glycoside (baicalin) and organic acids (chlorogenic acid and caffeic acid) respectively, and a combined evaporative light scattering detector was used to detect three steroid compounds (cholic acid, ursodesoxycholic acid and hyodeoxycholic acid). The proposed method permitted the simultaneous separation and determination of five groups of compounds in Qingkailing injection, and acceptable validation results of the precision, repeatability, stability and accuracy tests were achieved. The method was applied to the analysis of 19 Qingkailing injection samples from three different plants, and the results indicated that the method could be used as a convenient and reliable method in the multi-component determination and quality control of traditional Chinese medicines. PMID- 16350791 TI - [Analysis of recombinant erythropoietin and its tryptic digest by capillary electrophoresis and capillary electrophoresis-electrospray ionization-mass spectrometry using capillary coated with 6,6-ionene]. AB - The microheterogeneity of recombinant human erythropoietin (rhEPO) was analyzed by capillary electrophoresis (CE) using a capillary coated with 6,6-ionene. The applicability of a volatile electrolyte for fast analysis of tryptic fragments of rhEPO with online CE-electrospray ionization-mass spectrometry (ESI-MS) was investigated, resulting in a reproducible separation of eleven rhEPO tryptic fragments within 22 min under the following conditions: running buffer 300 mmol/L acetic acid-ammonium acetate (HAc-NH4Ac), pH 4.80, separation voltage -15 kV and capillary temperature 25 degrees C. The proposed method is rapid and effective, and can be used for the structural analysis of related proteins. PMID- 16350792 TI - [Determination of polybrominated diphenyl ethers in sewage sludge by high resolution gas chromatography coupled with high resolution mass spectrometry]. AB - Polybrominated diphenyl ethers (PBDEs) are a group of brominated flame retardants, which are manufactured in large quantities and widely used in a variety of consumer goods. Recently they spread ubiquitously as environmental contaminants. In order to investigate the pollution level of PBDEs in different environmental samples, a method has been established by using high resolution gas chromatography coupled with high resolution mass spectrometry (HRGC-HRMS). The method was used to analyze biosamples obtained from the international inter calibration. The results were satisfactory by analyzing z-score figures. Then, PBDE contents in three samples of sewage sludge were investigated by the method after Soxhlet extraction and multi-layer silica gel chromatographic separation. The number of PBDEs determined was 19. From the results, we can see that the contamination level of PBDEs in the sample from Beijing was higher than those from two other areas. The method was validated and applied for the analysis of environmental samples. PMID- 16350793 TI - [Determination of amygdalin in hawthorn by high performance liquid chromatography]. AB - A suitable method for extraction of amygdalin from hawthorn has been established. At first, the lipophilic components were removed with petroleum ether by ultrasonic extraction. The amygdalin was then extracted by methanol in a Soxhlet's apparatus. For quantitation, a high performance liquid chromatographic method was developed by using a reversed-phase C18 column, mobile phase of methanol-water (15:85, v/v) and a detection wavelengh of 215 nm. It can be concluded that the content of amygdalin is higher in the seeds than that in the hawthorn powder without the seeds and the yield of amygdalin is higher in the hawthorn pieces than that in the hawthorn powder. PMID- 16350794 TI - [Preparation and evaluation of organic-inorganic hybrid silica-based monolithic columns for capillary electrochromatography]. AB - A new type of organic-inorganic hybrid porous silica-based monolithic columns was developed for reversed-phase capillary electrochromatography (RP-CEC). The monolithic columns were prepared from in situ co-condensation of tetraethoxysilane (TEOS) and phenyltriethoxysilane (PTES) precursors via a two step catalytic sol-gel procedure in the capillary to create phenyl groups distributed throughout the silica matrix for chromatographic interaction. The functionalized monolithic stationary phase was created without additional surface derivatization. The reaction conditions for the preparation of the monolithic columns were optimized. The structure and pore size distribution of the monolithic columns were characterized by scanning electron microscopy and mercury porosimetry, respectively. The effect of water concentration on the column structure was studied, and the influence of the ratio of two precursors on the pore size distribution was also investigated. The retention behavior of polycyclic aromatic hydrocarbons on the monolithic column is discussed. Seven phenols were also separated with an average efficiency over 100 000 plates/m. PMID- 16350795 TI - [Synthesis, characterization and evaluation of octadecyl ester-bonded monolithic column]. AB - An ester-bonded monolithic silica column for reversed-phase high performance liquid chromatography was synthesized by reacting stearic acid with gamma glycidoxypropyltrimethoxysilane and coupling the intermediate product onto monolithic silica column. Characterization of the prepared column was carried out with Fourier transformation infrared spectrometry and elemental analysis. The chromatographic properties of the packing were evaluated by using benzene, biphenyl, and anthracene as samples and methanol-water as binary mobile phase. The column back-pressure and separation effect were also evaluated for fast separation. The results show that the stationary phase has satisfactory chromatographic properties and resists hydrolysis between pH = 2 - 8, and it can be used for the fast-separation of solutes efficiently. PMID- 16350796 TI - Antibodies as tailor-made chiral selectors: an interdisciplinary approach to enantiomer separation and detection. AB - It has long been known that the configurational isomers of biologically active compounds, e.g., nutrients, pesticides, and drugs, may exhibit different activities in a chiral environment such as the human body. Although the majority of drugs presently in development are chiral, analytical and preparative methods for the quantitative determination and purification of stereoisomers still lag behind. One reason is that commonly used chiral selectors for the direct resolution of enantiomers are not tailor-made for a specific analyte. The identification of suitable selectors for a particular pair of enantiomers still requires considerable experimentation and is generally demanding with regard to material, time and labor. The rational design of chiral host molecules, therefore, represents a challenge in facilitating enantiomer analysis. In this article, we describe how a combination of techniques ranging from organic synthesis to molecular biology yields antibodies of predetermined specificity and stereoselectivity that can be used as tailor-made chiral selectors for the chromatographic separation of enantiomers and their sensitive detection in immunosensors. PMID- 16350797 TI - [Determination of osteopontin at trace levels by non-gel sieving capillary electrophoresis]. AB - A method of non-gel sieving capillary electrophoresis (NGSCE) was established to determine osteopontin at trace levels. The capillary used was uncoated fused silica with a size of 57 cm x 75 microm i. d. and an effective length of 50 cm. The electrode buffer was a 150 mmol/L boric acid-borate buffer containing 30 g/L polyethylene glycol 20000 (pH 10.0). Other conditions were as follows: separation voltage 23 kV; detection wavelength 214 nm; pressure of injecting sample 3.4 kPa (0.5 psi) x 5 s; and column temperature 25 degrees C. The NGSCE method had excellent linearity with correlation coefficient of 0. 996, and reproducibility with the relative standard deviation of migration time of osteopontin less than 5% . The recovery was 95% and better, the sensitivity was 0. 079 g/L. Osteopontin secreted by vascular smooth muscle cells was determined by the NGSCE method at different times after serum withdrawal, and the results were in agreement with those of Western blot method. The results indicate that NGSCE is a simple and rapid method of determining osteopontin at trace levels. This method only needs a micro-amount of sample and is easily automated. PMID- 16350798 TI - [Determination of bioactive components in Semencustae by capillary electrophoresis with electrochemical detection]. AB - A method of high performance capillary electrophoresis with electrochemical detection (CE-ECD) has been developed for the determination of five bioactive components in Semencustae, namely rutin, hyperoside, kaempferol, p-coumaric acid and quercetin. The effects of several factors such as the acidity and concentration of the running buffer, the separation voltage, the applied potential and the injection time on CE-ECD were investigated. Under the optimized conditions, these five components can be separated in a 50.0 mmol/L borax running buffer (pH 9.0) within 19 minutes. A 300 microm diameter carbon disk electrode was used as the working electrode positioned carefully opposite to the outlet of the capillary in a wall-jet configuration at potential of + 950 mV (vs. saturated calomel electrode as reference electrode, SCE). Good linear relationships were established between the peak current and concentration of analytes over two orders of magnitude. The detection limits (S/N = 3) were 1.93 x 10(-5), 3.55 x 10(-4), 3.65 x 10(-5), 1. 73 x 10(-5) and 1.46 x 10(-4) g/L for rutin, hyperoside, kaempferol, p-coumaric acid and quercetin, respectively. The method has been successfully applied to the determination of these analytes in Semencustae samples after a relatively simple extraction procedure, and the assay results were satisfactory. PMID- 16350799 TI - [Determination of isosorbide-5-nitrate in human plasma by reversed-phase high performance liquid chromatography]. AB - The method was established to determine the level of isosorbide-5-nitrate in human plasma by reversed-phase high performance liquid chromatography (RP-HPLC). The sample preparation was carried by alkalizing the plasma sample followed by extraction with dichloromethane. The HPLC analysis was performed under the conditions as follows: a mixture of an aqueous buffer (pH adjusted to 7.8 by 0. 03 mol/L ammonia water) and acetonitrile (80: 20, v/v) as mobile phase, paracetamol as the internal standard, and the detection at 230 nm. The linear range was 20 - 1 000 microg/L using the ratio of peak areas; the detection limit was 12 microg/L; the average recovery was (97.11 +/- 2.45)% - (104.34 +/- 2.17)%, the intra-day relative standard deviations (RSDs) were less than 2.52%, and inter day RSDs were less than 5.21%. PMID- 16350800 TI - [Determination of lactone components in chuanxiong by reversed-phase high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method was developed for the simultaneous determination of four main lactone components including senkyunolide-H, senkyunolide-I, sedanenolide and Z-ligustilide in Chuanxiong, a Chinese herbal medicine. The samples were pretreated using ultrasonic extraction, and 80% ethanol was used as extractant in the experiment. The chromatographic analysis was carried out using an XDB-C8 column and the mobile phase consisted of methanol-1% (volume fraction) acetic acid water (55:45-->100:0, v/v, in 15 min). The flow rate was 0. 8 mL/min. A diode array detector was used to detect the compositions and 280 nm was chosen as the detection wavelength. The whole process could be performed within 15 min. The operating curves were found to be linear over the ranges of 0.4 - 2.2 mg/L (r=0. 9992), 0.4 -2.2 mg/L (r=0.9979), 2.6 -7.8 mg/L (r=0. 9992) and 4.8 14.3 mg/L (r = 0.9998) and the detection limits (S/N = 3) were 0.024, 0.027, 0.018 and 0.011 microg for senkyunolide-H, senkyunolide-I, sedanenolide and Z-ligustilide, respectively. The recoveries of the four lactone components were found in the range of 96% - 108%. The rapid and accurate method has been successfully applied to the simultaneous determination of the four lactone components in Chuanxiong. PMID- 16350801 TI - [Isolation, purification and identification of geniposidic acid in Eucommia ulmoides oliv. by silica column chromatography, reversed-phase high performance liquid chromatography, liquid chromatography-electrospray ionization mass spectrometry and nuclear magnetic resonance]. AB - The methods using silica column chromatography, reversed-phase high performance liquid chromatography, liquid chromatography-electrospray ionization mass spectrometry and nuclear magnetic resonance (RP-HPLC/LC-ESI-MS/NMR) for the isolation, purification and identification of geniposidic acid (GPA) in Eucommia ulmoides Oliv. were established. The crude extract of E. ulmoides was obtained after E. ulmoides was extracted with 70% ethanol and the extract was concentrated under reduced pressures. The obtained crude extract dissolved in methanol was injected to a silica gel column and eluted with chloroform-methanol. The eluted fractions were analyzed by RP-HPLC. The eluant from chloroform-methanol (8:1, v/v) was indicated to be a single component by HPLC with a retention time of 5.142 min. The HPLC analysis of the mixture of this fraction with a GPA standard resulted in a single peak. The ultraviolet and infrared spectra of the eluted solution were essentially the same to those of GPA. On the other hand, LC-ESI-MS, 1H- and 13C-NMR experiments also indicated that the fraction was identical to GPA. PMID- 16350802 TI - [Simultaneous determination of water-soluble vitamins C, B1, B2 and B6 in almonds by high performance liquid chromatography]. AB - A method has been developed for the simultaneous determination of four water soluble vitamins C, B1, B2 and B6 in almond by high performance liquid chromatography. The contents of vitamins C, B1, B2 and B6 in almonds in Pucheng County, Shaanxi Province were determined under optimized conditions as follows. In the first step, the sample was prepared by acid hydrolysis. In the second step, the separation was performed on an Inertsil ODS-3 column (25 cm x 4.6 mm i. d. , 5.0 microm) with a mobile phase of 0.05 mol/L KH2PO4 (pH 6.0)-methanol (70: 30, v/v) and a detection wavelength at 265 nm. The linear ranges of four vitamins were within 5.0 - 50.0 mg/L (r = 0. 999 0 - 0. 999 7). At the spike level ranged between 5.0 - 20.0 mg/kg, the average recoveries of the four vitamins in the Pucheng almond ranged from 91.77% to 99.30% with relative standard deviations between 0.31% and 1.98%. For vitamins B2 and B1, the contents were found to be 4.27 - 4.53 mg/kg and 0.799 - 0.838 mg/kg, respectively, while vitamins C and B6 were not discovered in Pucheng almonds. The method is simple, rapid, reproducible and accurate. PMID- 16350803 TI - [Simultaneous determination of sudan red dyes in foods by high performance liquid chromatography with a clean-up procedure by gel column]. AB - A method was developed for the simultaneous determination of Sudan Red I, II, III and IV in foods by high performance liquid chromatography (HPLC) with a clean-up procedure by gel column. Sample was extracted from foods with ethanol. The extract was cleaned up with a Bio-Beads SX3 gel column (200 mm x 10 mm i. d. ) and eluted with cyclohexane-ethyl acetate (1:1, v/v). The analysis was performed on a Symmetry Shield RP18 column (250 mm x 4. 6 mm i. d. , 5 microm) with 100% methanol as the mobile phase at a flow rate of 1.5 mL/min, detection at 478 nm and confirmation by diode-array spectra. All of the four compounds demonstrated good linear relationship (r > 0.999) in the range of 0.1 - 10.0 mg/L. The limits of detection (LOD) were from 7 to 14 microg/kg. The average recoveries for all four dyes (spiked at the levels of 0.25 and 2. 5 mg/kg) in chili sauce and sausage ranged from 80.7% to 96.3%, and the relative standard deviations were from 2.4% to 5.9%. The method is sensitive, reliable and can be applied for the analysis of four Sudan Red dyes in foods. PMID- 16350804 TI - [Determination of phenylarsenic compounds in environmental samples by high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method was developed for simultaneous determination of phenylarsonic acid (PAA), phenylarsine oxide (PAO), diphenylarsinic acid (DPAA), bis (diphenylarsine) oxide (BDPAO) and triphenylarsine (TPA) in soil samples. The extraction solvent for soil samples was acetonitrile and ultrasound was used in the extraction. Good separation was achieved by using an Inertsil CN-3 column (250 mm x 4.6 mm i.d., 5 microm) with the following conditions: gradient program of mobile phase, a 10 min linear gradient from 100% A (a mixture of acetonitrile-2.5 g/L KH2PO4 (pH 2.0) buffer (5:95, v/v) ) to 100% B ( a mixture of acetonitrile-2. 5 g/L KH2PO4 (pH 2.0) buffer (50:50, v/v)), and isocratic at 100% B from 10.01 min to 28 min; flow rate of 1.0 mL/min; time-programmed UV detection at 210 nm (0 - 10.5 min, 20 - 28 min) and 268 nm (10. 5 - 20 min). The linear range of PAA, PAO, DPAA, BDPAO and TPA were 8 - 30, 5 - 40, 20 -4000, 120 - 8000 and 1 - 60 mg/L, respectively. The detection limits at a signal-to-noise ratio of 3 were 0.1, 0.1, 0.2, 10, and 0.1 mg/L for PAA, PAO, DPAA, BDPAO and TPA, respectively. PMID- 16350805 TI - [Analysis of volatile oil of garlic by gas chromatography-mass spectrometry]. AB - The volatile oil of garlic was extracted by hydrodistillation method and gas chromatography-mass spectrometry was applied to analyse the compounds in the oil. The best extraction conditions for high-content, effective components were obtained through optimization. The capillary column was an HP-5MS column (25 mm x 0.25 mm i.d. x 0.25 microm); oven temperature increased with a rate of 5 degrees C /min from 80 to 300 degrees C, and then maintained for 20 min; sample size of 1 microL; split ratio of 100:1; carrier gas of helium (1 mL/min). Mass spectra were obtained at 70 eV. The temperatures of injector base, ionization source were maintained at 270 degrees C, 230 degrees C respectively. Under these conditions, twenty compounds in the volatile oil of garlic were isolated and identified, and the content of each was determined. Sulfur-containing compounds were found to be the principal components, of which the major compound was diallyl trisulfide with the content of more than 30%, which is higher than the others in the literature. The experimental results also indicated that hydrodistillation method is an effective method for officinal component extraction. In addition, it was also demonstrated that the garlic volatile oil was stable when stored at 0 degrees C for 6 months. PMID- 16350806 TI - [Determination of the dissociation constants of basic compounds by reversed-phase high performance liquid chromatography on C12-zirconia]. AB - The high performance liquid chromatographic (HPLC) behavior of aromatic amines and pyridine derivatives was studied on a new reversed-phase stationary phase, C12-alkyl-modifled zirconia. The effects of mobile phase variables such as methanol content and pH were investigated. The results showed that the stationary phase was stable in the mobile phase with a pH range of 2 - 12. It was found that on the new stationary phase the bases demonstrated the "traditional" behavior of retention due to their hydrophobicity. The retention factors of these basic compounds on the stationary phase increased rapidly with pH from pH 2 -7, and then increased slowly until reaching a constant value. The dissociation constants (pKa) of 13 basic compounds were determined based on the relationship between their retention factors and the pH of mobile phase. Compared with references, the errors of the pKa values obtained by this HPLC method were within 0.35 pH units. PMID- 16350807 TI - Thin layer chromatographic study of bauxite and quantitative estimation of co existing Al3+, Fe2+ and Ti4+. AB - Thin layer chromatography in combination with spectrophotometry and titrimetry has been used to evaluate chromatographic characteristics of bauxite constituents. The retention behaviors of four major constituents (Al3+, Fe2+, Ti4+, Si4+) in bauxite mineral have been examined on plain and modified layers of silica gel G, silica gel H and cellulose with mobile phases containing aqueous sodium chloride, formic acid and hydrochloric acid. Ternary separation of Al-Fe Ti was achieved on chromatographic plates made of silica gel H. The pH effect and presence of impurity elements in samples, nature of stationary phases on the ternary separation and detection limits of bauxite constituents were studied. Silicon in bauxite was detected on cellulose plates. Quantitative determinations of Al3+, Fe2+ and Ti4+ on silica gel H impregnated with sodium formate layers were achieved by titrimetry and spectrophotometry. PMID- 16350808 TI - [Analysis of dodecyl glucoside by RP-HPLC]. PMID- 16350809 TI - [Determination of aripiprazole by nonaqueous reversed-phase high performance liquid chromatography]. PMID- 16350810 TI - [Determination of cytidine triphosphate and related compound triphos and related compounds in the liquid of enzymatical reaction by high performance liquid chromatography]. PMID- 16350811 TI - [Determination of muscone in Xingnaojing injection by gas chromatography with solid phase microextraction]. PMID- 16350812 TI - [Simultaneous determination of several dye intermediates in oxidation hair dyes by gas chromatography-mass spectrometry]. PMID- 16350813 TI - Association between attributed cause of end-stage renal disease and risk of death in Brazilian patients receiving renal replacement therapy. AB - BACKGROUND: Studies conducted in several countries have indicated that the survival of patients undergoing renal replacement therapy (RRT) depends on the attributed cause of end-stage renal disease (ESRD). OBJECTIVES: This study was conducted to evaluate the association between attributed cause of ESRD and mortality risk in RRT patients in Brazil. METHODS: We analyzed 88,881 patients from the Brazilian Ministry of Health Registry who were undergoing RRT between April 1997 and July 2000. Cox proportional hazards models were used to estimate the relative risk (RR) of death in patients with ESRD secondary to diabetes mellitus (DM), polycystic kidney disease (PKD), and primary glomerulopathies (GN) compared with a reference group comprised of patients with ESRD caused by hypertensive nephropathy. Patient's age, gender, and length of time (years) in RRT before inclusion in the registry (vintage) were included in the adjusted Cox model. RESULTS: Compared with the reference group, the mortality risk was 27% lower in patients with PKD (RR=0.73, 95% CI: 0.65-0.83, p<0.0001); 29% lower in patients with GN (RR=0.71, 95% CI: 0.68-0.74, p<0.0001); and 100% greater in DM patients (RR=2.00, 95% CI: 1.92-2.10, p<0.0001). These relative risks remained statistically significant after adjustment for age, gender, and length of time in RRT before inclusion in the registry. CONCLUSIONS: Our data indicate that compared with the patients with hypertensive nephrosclerosis as attributed cause of ESRD, patients undergoing RRT in Brazil with idiopathic glomerulopathy and polycystic kidney disease have a lower risk of mortality, and patients with diabetes mellitus have a greater risk of mortality. PMID- 16350814 TI - Urinary tuberculosis: ten years' experience. AB - In this study of 26 patients, clinical features diagnosed as urinary tuberculosis in our nephrology and urology clinics between 1993 and 2002 were investigated retrospectively. Fifteen patients (52%) were male, and mean age was 43.5 (18-71). Twenty percent of the patients were asymptomatic. Frequency-dysuria (46%), flank pain (33%), and macroscopic hematuria (12%) were presenting symptoms. Physical examination was not diagnostically helpful in most patients. Hematuria and/or pyuria were detected in 80% of the patients. Eleven patients had positive urine cultures of Mycobacterium tuberculosis (42%), and 7 patients had positive smears (25%). Definitive diagnosis of urinary tuberculosis was established microbiologically in 15 patients (58%) and histopathologically in 11 patients (42%). Tuberculin skin test was positive in 60% of the patients. Eight patients had an abnormal chest roentgenogram. Hydronephrosis (majority bilateral) in 11 patients (42%), contracted bladder in 9 patients (34.6%), and renal calcification in 6 patients (23%) were detected. Two patients also had genital tuberculosis (epididymoorchitis). Although only medical treatment was applied in 13 patients for 9 months, in the rest of the patients medical therapy plus surgical intervention was carried out. End-stage renal failure developed in one patient who died on hemodialysis. Renal functions had decreased moderately in two other patients. In conclusion, the diagnosis of urinary tuberculosis was able to be established after the obstructive complications and functional losses were developed in a fair number of cases. Surgical treatment was carried out in half the patients. Urinary tuberculosis should be taken into consideration because early diagnosis and treatment is very important for the presenting of irreversible sequelae. PMID- 16350815 TI - Prognostic value of acute physiology and chronic health evaluation II and organ system failure in patients with acute renal failure requiring dialysis. AB - BACKGROUND: Despite advances in modern technology of dialysis, prognosis of patients with acute renal failure (ARF) remains poor. To give the clinicians the most useful information, a model that accurately predicts outcome early in the course of ARF is required. However, because ARF is a heterogeneous syndrome and occurs in patients with diverse etiologies and some coexisting diseases, predicting outcome early is hard. The aim of this study is to evaluate prospectively the Acute Physiology and Chronic Health Evaluation (APACHE II) and organ system failure (OSF) models, evaluated prior to dialysis, in predicting hospital mortality. METHODS: From June 2002 to March 2004, ARF patients requiring dialysis at Chang Gung Memorial Hospital, Chiayi, were prospectively recruited for this study. The worst clinical and laboratory data in the 24 hours before initiation of dialysis were prospectively evaluated, and the patients' APACHE II score and OSF number were assessed. RESULTS: A total of 61 patients (40 male and 21 female) were enrolled, of whom 38 (62.3%) died before discharge. By multivariate logistic regression, the APACHE II score (odds ratio 1.3 per increase in one score; P<0.001), or OSF number (odds ratio 1.9 per increase in one OSF; P<0.01) and oliguria (odds ratio 4.2; P=0.04), were found to be statistically significant prognostic factors for hospital mortality. Mortality increased progressively and significantly as OSF number (chi-square for trend; P=0.001) or the APACHE II score (chi-square for trend; P < 0.001) increased. By using Youden's index, the best cut-off value for APACHE II was 24, with 63% sensitivity and 96% specificity. The best cut-off value for OSF number was 2, with a sensitivity of 81.6% and a specificity of 60.9%. The areas under the receiver operating characteristic curves for APACHE II and OSF number were 0.847 (95% confidence interval (CI)=0.752-0.942; P<0.01) and 0.769 (95% CI=0.646-892; P<0.001), respectively, indicating good model discrimination. CONCLUSIONS: This study concludes that APACHE II and OSF number measured prior to initiation of dialysis reliably predict outcomes of ARF patients requiring dialysis. The mortality rates increase as the APACHE II score or OSF number increases. For predicting mortality, the APACHE II score > or = 24 was found to have 63% sensitivity and 96% specificity, and OSF number> or = 2 had 81.6% sensitivity and 60.9% specificity. PMID- 16350816 TI - Transforming growth factor-beta1 gene polymorphism in renal transplant recipients. AB - BACKGROUND: Cytokine transforming growth factor (TGF) is involved in regulation of tissue repair after injury. More recently, TGF-beta1 codon 10 gene polymorphism has been shown to be associated with circulating TGF-beta levels. We tested whether TGF-beta1 genotype polymorphism was predictive of renal allograft function decline. PATIENTS AND METHODS: The study population consisted of 129 consecutive cadaveric or living related renal transplant recipients at our center between 1985 and 2001. The recipient TGF-beta1 genotype polymorphism was determined from peripheral blood leucocytes DNA. The primary endpoint was rate of glomerular filtration rate decline between the first year and the third year of transplant. RESULTS: Baseline glomerular filtration rate as estimated by MDRD study equation at 1 year measured 50+/-17 mL/min/1.73 m2. At the end of the 3 year follow-up period, 52 patients (40%) experienced biopsy-confirmed acute rejections. Frequency and severity of allograft rejection did not differ with TGF beta genotypes. However, the decline in glomerular filtration rate was significantly greater in Leu/Leu (TT) than Leu/Pro (CT) recipients, 6.3+/-16.9 mL/min/1.73 m2 versus 0.1+/-10.2 mL/min/1.73 m2, p=0.04. CONCLUSION: Our results demonstrate that recipient TGF-beta1 codon 10 Leu/Leu homozygosity is a potential risk factor of kidney allograft function decline. PMID- 16350817 TI - Ultrasonic gallbladder function in chronic kidney disease: does predialysis, hemodialysis, or CAPD affect it? AB - BACKGROUND: There are contradictory reports about the prevalence of cholelithiasis in chronic kidney disease (CKD). The pathogenesis of gallstones is associated with the lithogenic changes of bile composition, increased tendency to nucleation, and decreased gallbladder motility. The studies related to these factors can predict the development of cholelithiasis. The aim of this study was to evaluate the ultrasonic gallbladder function in CKD and to compare it in predialysis (PreD), hemodialysis (HD), and continuous ambulatory peritoneal dialysis (CAPD) patients. METHODS: Age, gender, and body mass index matched 49 CKD patients (14 PreD, 19 HD, 16 CAPD), and 17 control individuals were included in the study. Diabetic and cirrhotic patients were not included. Ultrasonic gallbladder volume was evaluated in pre- and postprandial period, and ejection fraction was calculated. We also measured several biochemical parameters (cholesterol, triglyceride, blood urea nitrogen (BUN), creatinine, calcium, Phosphorus, parathormone, albumin, total protein) in blood. RESULTS: Preprandial gallbladder volume in PreD, HD, CAPD, and control groups were 26.7 +/-13.6, 20.8+/-10.4, 23.2+/-14.7, and 26.4+/-14.8 mL, respectively (p > 0.05). Ejection fractions were 54.1 +/- 22.9%, 54.9 +/- 23.9%, 48.6 +/- 15.9%, and 51.8 +/- 19.2% in PreD, HD, CAPD, and control groups, respectively (p > 0.05). Serum triglyceride was higher in PreD patients than control group (207 +/- 144 vs. 110 +/-48 mg/dL) (p<0.05). Serum BUN, Cre, P, and PTH levels were higher in CKD groups than the control group, whereas serum total protein and albumin levels were higher in the control group (p<0.05). Serum Ca was lower in PreD and HD patients than in the controls (p<0.05). CONCLUSIONS: In conclusion, CKD and renal replacement therapy (HD and CAPD) do not affect gallbladder functions, but more studies are needed to evaluate prevalence of gallstones, gallbladder motility, and the composition of bile in CKD. PMID- 16350818 TI - The value of computed tomography-derived coronary artery calcification score in coronary artery disease detection in asymptomatic hemodialysis patients. AB - BACKGROUND: We evaluated the value of coronary artery calcification (CAC) score in coronary artery disease (CAD) detection in asymptomatic hemodialysis (HD) patients by evaluating the association among CAC score, exercise electrocardiography (EECG), and Thallium-201 dipyridamole scintigraphy. Correlation between aortic pulse wave velocity (PWV) and CAC score was also evaluated. METHODS: CAC score was assessed with conventional computed tomography in 40 patients. Thirty patients completed EECG and 25; those with a positive CAC score and/or a positive EECG performed Thallium dipyridamole scintigraphy. Carotid-femoral PWV was assessed in all patients. RESULTS: There was no association among CAC score and EECG or Thallium dipyridamole scintigraphy. In contrast, CAC score was correlated with aortic PWV. CONCLUSION: The previous results question the role of CAC score in the detection of CAD in asymptomatic HD patients. The correlation between CAC score and aortic PWV raises the possibility that CAC score represents more an indicator of coronary artery medial wall calcification than a marker of CAD. PMID- 16350819 TI - DNA oxidative damage in patients with dialysis treatment. AB - BACKGROUND/AIMS: Chronic renal patients on hemodialysis (HD) and peritoneal dialysis (PD) treatment are exposed to oxidative stress and DNA damage. The objective of this study was to assess the oxidative damage to DNA in end-stage chronic renal failure, before and after vitamin E supplementation. METHODS: Patients on HD (n=29) and PD (n=22) received oral supplementation with 300 mg vitamin E three times a week for 4 weeks. A blood sample was collected at the beginning and at the end of the supplementation cycle for the determination of vitamin E levels (high-performance liquid chromatography), carbonyl groups, and DNA damage (8-hydroxy 2'-deoxyguanosine [8-OHdG] and comet assay). RESULTS: After supplementation, vitamin E concentration was increased by about 50%. Protein oxidation was initially observed in both groups, with a reduction after supplementation. DNA damage detected by the comet assay and by 8-OHdG analysis was significantly reduced (p<0.05) after supplementation in both groups. CONCLUSIONS: Vitamin E supplementation reduced oxidative DNA damage in both HD and PD patients. Treatments such as HD and PD induce oxidative stress and consequent DNA damage, and increased plasma vitamin E levels significantly contribute to the normalization of these events. PMID- 16350820 TI - Relation between depression, some laboratory parameters, and quality of life in hemodialysis patients. AB - Depression is common in patients with end-stage renal disease (ESRD) and is associated with increased mortality and morbidity. Several investigators have estimated that depression occurs in about 20% to 30% of dialysis patients. The aim of this study was to investigate the relationship between depression, some laboratory parameters, and quality of life (QOL) in hemodialysis patients. Forty three hemodialysis patients (mean age 40.5+/-15.2; M=28, F=15) were included in the study. Hamilton Depression Scale (HAMD), Hamilton Anxiety Scale (HAMA), and short form with 36 (SF-36) were used for evaluation. Subsequently, patients were divided into two groups according to HAMD scores: group 1, those who had a low HAMD score (between 0 and 7), and group 2, those who had a high HAMD score (over 7). The two groups were compared in terms of anxiety scores, QOL scores, and some laboratory parameters. The group 2 patients (n=21; M= 13, F=8) had lower levels of hemoglobin than the group 1 patients (9.5+/-1.7 vs. 10.7+/- 1.4 g/dL, respectively; p<0.01). Group 2 patients also had lower SF-36 scores than group 1 patients (91.5+/-21.3 vs. 74.9+/- 13.6, respectively; p=0.03). On the contrary, the patients of group 2 had higher HAMA scores than group 1 patients (16.6+/-6.9 vs. 6.3+/-3.5, respectively; p<0.01) and CRP level (10.7+/-4.6 vs. 4.5+/-3.8, respectively; p<0.001). A significant correlation was found between depression scores and C-reactive protein (CRP) (r= 0.57, p < 0.001) and HAMA scores (r= 0.43, p<0.05). In contrast, a negative correlation was found between HAMD scores and albumin (r=-0.43, p<0.05), hemoglobin (r=-0.38, p=0.015) and SF-36 scores (r=0.39, p=0.032). These findings demonstrate that there is a relationship among high depression score, low levels of hemoglobin and albumin, high CRP level, low SF-36 score, and high anxiety score. Evaluation of psychiatric status should be part of the care provided to hemodialysis patients. PMID- 16350821 TI - Progression of kidney damage in Balkan endemic nephropathy: a 15-year follow-up of patients with kidney biopsy examination. AB - Progression of kidney damage was studied in 18 patients with Balkan endemic nephropathy (BEN), with a mean 15-year follow-up after renal biopsy. According to kidney function, estimated by 99mTc-DTPA clearance, patients were divided into three groups: with apparently normal kidney function (clearance 103.5+/-21.3 mL/min/1.73 m2), with incipient renal failure (clearance 65.5 +/- 11.3), and with advanced renal failure (clearance 28.0+/-6.2). The mean yearly decrease of glomerular filtration rate was 2.74 mL/min. In two patients, an increase of kidney function was recorded. Six patients become dialysis dependent, two from the group with incipient renal failure, but all four from the group with advanced renal failure. Three patients died after 8 to 12 years of follow-up, one from causes unrelated to kidney disease and two from end-stage renal failure. This study has shown that BEN is characterized by a slow course and prolonged evolution, modified by medical supervision and treatment. PMID- 16350822 TI - Intravenous iron gluconate administration increases circulating PAPP-A in hemodialysis patients. AB - BACKGROUND: Pregnancy-associated plasma protein-A (PAPP-A) is a proatherosclerotic molecule, interrelated with oxidative stress in hemodialysis (HD) patients. As intravenous (IV) iron might enhance oxidative stress in HD patients, this study investigates circulating PAPP-A during HD session and after IV iron administration. METHODS: In 20 HD patients, plasma PAPP-A concentration was assessed immunochemically during 2 HD sessions (prior to HD and at 60, 130, and 240 min of HD session). Sodium ferric gluconate (62.5 mg) was given IV to all patients 65 min after the start of the second HD. RESULTS: Sixty-five min after IV iron application, there was a significant increase in plasma PAPP-A (from 36.0+/-9.9 to 79.6+/-28.9 mU/L, p<0.0001). At the end of this HD session, PAPP-A decreased significantly (p<0.0001), but still remained 1.5-fold greater compared with predialysis levels (p<0.0005). CONCLUSION: IV iron increases circulating PAPP-A, and in this way, it might contribute to more pronounced cardiovascular complications in HD patients. PMID- 16350823 TI - Combination of duplex ultrasound-guided manual declotting and percutaneous transluminal angioplasty in thrombosed native dialysis fistulas. AB - BACKGROUND: To investigate the safety, feasibility, efficacy, and long-term patency rate of manual declotting under duplex ultrasound (US) guidance followed by percutaneous transluminal angioplasty (PTA) in thrombosed native arteriovenous fistulas (AVFs). METHODS: Of 87 consecutive thrombosed AVFs evaluated by duplex US, 22 patients with 25 recently thrombotic events in 22 AVFs were suitable for manual declotting. PTA was performed following successful declotting, and long term patency was assessed. RESULTS: The procedure success rate of manual declotting was 80% (20 of 25), and a residual stenosis of 74+/-9% was identified by duplex US after declotting. PTA reduced the diameter stenosis to 25+/-6% and increased the lumen diameter from 1.33 +/-0.85 mm to 4.62+/-0.98 mm. Neither embolic nor bleeding complications were noted during the procedure. The average procedure time and the fluoroscopy time were 28.4+/-9.9 and 7.2+/-4.1 minutes, respectively. Primary patency rates at 1, 2, and 3 years were 47%, 35%, and 28%; assisted primary patency rates at 1, 2, and 3 years were 71%, 63%, and 63%; and secondary patency rates at 1, 2, and 3 years were 76%, 71%, and 63%, respectively, during a maximum follow-up period of 42 months. CONCLUSION: The combination of duplex US-guided manual declotting and angioplasty of underlying stenosis is a safe and feasible method to treat recently thrombosed native AVFs in selected patients. It simplifies the interventional procedure, reduces cost and radiation exposure time, and extends life span of dialysis fistula with acceptable long-term patency rate. PMID- 16350824 TI - Risk of developing fibroadenoma with the use of cyclosporine A in renal transplant recipients. AB - To investigate the effect of cyclosporine A (Cyc A) on the development of fibroadenomas, 30 renal transplant patients and 20 chronic renal failure patients on dialysis were breast examined with ultrasonography and/or mammography. Of the renal transplant patients, 17 were receiving Cyc A-based combination therapy for immunosuppression. All patients were female with the age range of 29.7+/-9.2 years in the transplant group and 33.95+/-9.91 in the dialysis group. Eight of the 17 patients receiving Cyc A had fibroadenomas, 5 of them having bilateral lesions. None of the other patients, those on dialysis and on non-Cyc A combination therapy had fibroadenomas. A significant difference for fibroadenoma incidence in patients receiving Cyc A combination immunosuppression was found. PMID- 16350825 TI - Effect of radiocontrasts on selected membrane transport systems. AB - BACKGROUND: This study was aimed at examining the effects of radiocontrast agents on selected membrane transport functions. METHODS: The effect of diatrizoate sodium (DS), diatrizoate meglumine (DM), and diatrizoate compound (DC) on Na+/K+ pump activity and number, L-arginine, and choline transports were evaluated in erythrocytes of normal individuals and patients undergoing cardiac catheterization. RESULTS: Norrmal individuals' erythrocytes potassium influxes were 1.50+/-0.35, 1.32+/-0.37, 1.28+/-0.30, and 1.01+/-0.25 mmol/L cell/h in control, DS, DM, and DC groups, respectively (P=0.004; DC vs. control). Patients exposure to Hypaque M-76 significantly diminished Na+/K+ pump activity (1.40+/ 0.36 before, vs. 1.27+/-0.40 mmol/L cell/h after; P=0.039). The number of Na+/K+ pumps was reduced (156+/-36 vs. 143+/-34 pumps/erythrocyte; P=0.015) in presence of DS. L-arginine and choline transports changed only at high radiocontrast concentrations. CONCLUSION: Selective changes in erythrocytes membrane transport function take place on exposure to radiocontrasts. PMID- 16350826 TI - Cardiac valve calcifications and left ventricular hypertrophy in hemodialysis patients. AB - Cardiac valve calcification (VC) is a common finding in end-stage renal disease patients. It was shown recently that VC is an independent predictor for all-cause and cardiovascular mortality in peritoneal dialysis patients. In hemodialysis (HD) patients, VC was associated with all-cause and cardiovascular mortality, but after adjusting for other cardiovascular risk factors and complications, as well as left ventricular mass index (LVMI), it lost significance. The aim of the study was to assess the relationship between VC and left ventricular hypertrophy in hemodialysis patients. Echocardiographic examination with mitral and aortic valves assessment and LVMI calculation was performed in 65 HD patients ages 49+/ 12, with duration of HD therapy 38+/-32 months. VC were found in 32 of 65 patients (49%)-Group VC(+), mitral valve calcifications (MVC) in 10, aortic valve calcifications (AVC) in 9, and both valves calcifications (MVC+AVC) in 13 patients. Patients with VC were older, on HD therapy were longer, had higher systolic and pulse pressure, and had higher LVMI. Patients with both VCs had the highest LVMI. No significant differences were found with respect to Ca, P, PTH, and mean Ca x P product, but the incidence of Ca x P product above 4.43 mmol2/L2 was higher in VC(+) compared with those without VCs. VC coexists with left ventricular hypertrophy, particularly when both valves are calcified. Even short lasting incidents of increased Ca x P product may lead to cardiac VC. PMID- 16350827 TI - Relation of thyroid disfunction, thyroid autoantibodies, and renal function. AB - Fifty-eight patients with thyroid dysfunction were included, comparing 27 healthy subjects to evaluate the relationship between thyroid autoimmunity, thyroid functional status, and renal function. Regardless of clinical status, hypothyroidism was defined as T3-T4upper limit of normal, and hyperthyroidism was defined as T3-T4>normal limits and TSH <0.1 mcU/mL. In all participants, serum antithyroperoxidase level, serum antithyroglobulin level, creatinine clearance (CC), and urinary albumin excretion rate (UAER) in 24-h urine collections were measured. Of 85 persons, 41 patients ages 20 to 71 years (median; 44) had hypothyroidism, 17 patients ages 22 to 56 years (median; 32) had hyperthyroidism, and 27 subjects ages 20 to 67 years (median; 50) were normal. Regarding thyroid autoantibody positiveness, the positive group had a higher UAER than the negative group (30.2 mg/day, 20.8 mg/day, respectively; p=0.05). In hypothyroid patients, UAER was significantly higher than in hyperthyroid patients and control subjects (30.1 mg/day, 11.8 mg/day, 10.5 mg/day, respectively; p<0.001). In the hypothyroid population, with regard to UAER and CC, we could not find a significant difference between the thyroid autoantibody positive and negative groups. In view of CC, hypothyroid patients had significantly lower CC than the hyperthyroid group (79.0 mL/ min, 86.5 mL/min, respectively; p<0.01). A careful control of the renal function in thyroid diseases should be evaluated. Renal dysfunction seems to be secondary to hypothyroidism rather than thyroid autoimmunity. PMID- 16350828 TI - Effects of thyroid function on the course of experimental chronic renal failure in rats. AB - Thyroid hormone has been reported to affect renal function. To investigate the effects of thyroid hormone on the progression of renal deterioration, thyroid hormone (dried thyroid) and an antithyroid drug (thiamazole) were administered to adriamycin (ADR)-induced renal failure rats. The rats were divided into four groups, including 1) ADR-DT, given dried thyroid and thiamazole; 2) ADR-T, given thiamazole; 3) ADR; and 4) control. The survival rate at the end of the study (22 weeks) was 62.5% in ADR-DT group and 100% in ADR-T, ADR, and control groups, respectively. There was a significant difference in the body weight and pulse rate between ADR-DT and ADR-T or ADR groups, except for the pulse rate at week 6 (P<0.05). The creatinine clearance was greater in the ADR-T group than in the ADR or ADR-DT groups at week 22, and was significantly different between the ADR-T and the ADR-DT groups (P<0.05). The fractional kidney weight and tubular changes were significantly greater in the ADR-DT group than in the ADR-T or ADR groups (P<0.05). The interstitial volume was significantly greater in the ADR-DT group than in the ADR-T group (P<0.05). We therefore conclude that a dried thyroid has an aggravative effect in the tubular changes and relative interstitial volume induced by ADR. PMID- 16350829 TI - Glutathione and glutathione peroxidase activities in blood of patients in early stages following kidney transplantation. AB - This study focuses on glutathione (GSH) level in red blood cells, as well as on glutathione peroxidases (GSH-Px) activities in red blood cells and in plasma of chronic renal failure (CRF) patients following renal transplantation. We want to focus our main attention on plasma GSH-Px, the selenoenzyme that is synthesized primarily in the kidney. In CRF patients, activity of this enzyme is significantly reduced, and the reduction decreases with the progress of the disease, reaching in the end-stage 20% to 30% of the activity of healthy patients. We have shown that following renal transplantation the activity of plasma GSH-Px is restored very rapidly, and 2 weeks after surgery it reached the value of the control group. Red blood cell GSH level is significantly higher in CRF patients, and following transplantation, no significant changes were observed. Red blood cell GSH-Px activity before transplantation was the same as in healthy patients and did not change significantly after surgery. PMID- 16350830 TI - Angiotensin II and endothelin in the renal cortex during the evolution of glycerol-induced acute tubular necrosis. AB - Hypertonic glycerol injection is one of the most frequently used models of experimental acute renal failure. Late structural changes such as interstitial fibrosis in the renal cortex and tubular atrophy have been detected after severe acute tubular necrosis (ATN). The aim of this study was to investigate the expression of angiotensin II (AII) and endothelin during the evolution of the ATN induced by glycerol and their relationships with the late structural changes observed in the kidneys. Forty-nine male Wistar rats were injected with a 50% glycerol solution, 8 mL/kg, divided into equal amounts, each administered into one hind leg, and 18 with 0.15 M NaCl solution. Blood and urine samples were collected 1, 5, 30, and 60 days after the injections to quantify sodium and creatinine; the animals were killed and the kidneys removed for histologic and immunohistochemical studies. The results of the immunohistochemical studies were scored according to the extent of staining in the cortical tubulointerstitium. Glycerol-injected rats presented a transitory increase in plasma creatinine levels and in fractional sodium excretion. The immunohistochemical studies showed increased AII and endothelin staining in the renal cortex from rats killed 5 days after glycerol injection (p<0.001) compared with control that persisted until day 60. The animals killed on days 30 and 60 also presented chronic lesions (fibrosis, tubular dilatation, and atrophy) in the renal cortex, despite the recovery of renal function. AII and endothelin may have contributed to the development of renal fibrosis in these rats. PMID- 16350831 TI - Effects of Hypoxis hemerocallidea (Fisch. & C.A. Mey.) [Hypoxidaceae] corm (African Potato) aqueous extract on renal electrolyte and fluid handling in the rat. AB - Current biomedical evidence suggests that Hypoxis hemerocallidea (Fisch. & C.A. Mey.) [Hypoxidaceae] (African Potato [AP]) corm extract may be useful in the management of type 2 diabetes mellitus. However, more recent reports have also indicated that certain herbal extracts attenuate the deterioration of kidney function in diabetes mellitus. Accordingly, this study investigated the effects of short- (acute) and long-term (chronic) administration of H. hemerocallidea corm aqueous extract (APE) on renal fluid and electrolyte handling in male Wistar rats. Acute effects of APE were investigated in separate groups of anesthetized rats challenged with a continuous jugular infusion of 0.077 M NaCl at 9 mL x h(- 1). After a 3.5-h equilibration period, consecutive 30-min urine collections were made over the subsequent 4 h of 1-h control, 1.5-h treatment, and 1.5-h recovery periods for measurements of urine flow, Na+, and K+ excretion rates. To establish the effects of acute APE, the extract was added to the infusate at doses of 90, 180, or 360 microg x h (-1) in separate groups of rats during the treatment period. For chronic studies, individually caged rats were administered twice with APE (30 mg x kg (-1) PO) every third consecutive day at 09h00 and 17h00 for 5 weeks. Control rats received distilled water (3 mL x kg(-1)). Urine volume and total urinary outputs of creatinine, Na+, and K+ were determined from 24-h samples. Acute infusion of APE produced a dose-dependent, significant (p < 0.01) decrease in urine flow, K+, and Na+ excretion rates. Chronic APE treatment significantly reduced urinary Na+ output between weeks 2 and 5, without affecting either urine flow or K+ excretion rates. When compared with control animals, APE significantly reduced GFR (2.54+/-0.09 mL x min (-1) vs. 1.52+/-0.02 mL x min ( 1)) and increased plasma creatinine concentration (55 +/- 3 micromol x L(-1) vs. 68 +/-6 micromol x L(-1)). The results from this study suggest that the H. hemerocallidea corm aqueous extract may impair kidney function. PMID- 16350832 TI - Methotrexate causes oxidative stress in rat kidney tissues. AB - AIM: Methotrexate (MTX), a folic acid antagonist, is one of the chemotherapeutic agents widely used in the treatment of some types of cancers. Nephrotoxicity is one of the complications of MTX treatment. The aim of this study was to investigate possible effects of MTX treatment on the oxidant/antioxidant status in rat kidney tissues and enzymatic mechanisms leading to nephrotoxicity. METHODS: For this aim, 10 Sprague-Dawley type female rats of 4 weeks old were used in the study. The animals were divided into two groups randomly. Five of them were used as control, and the others were treated with MTX intravenously (60 mg/m2 of body surface area per week) for 7 weeks. At the end of this period, they were sacrificed, and kidney tissues were removed to be used in the analyses of malondialdehyde (MDA) levels, antioxidant potential (AOP) values, and superoxide dismutase, catalase, glutathione peroxidase, xanthine oxidase, adenosine deaminase, and 5' nucleotidase enzyme activities. RESULTS: There was significant increase in the MDA level in the MTX group compared with the control group (1.74+/-0.23 nmol/mg vs. 1.04+/-0.30 nmol/mg; p<0.05, respectively). There were however no meaningful differences between enzyme activities and AOP values of the groups. CONCLUSION: It has been suggested that MTX leads to oxidative stress in rat kidney tissues, which might be one of the reasons for MTX-induced nephrotoxicity. PMID- 16350833 TI - The effects of mibefradil, a T-type Ca2+ channels blocker, on the renal dysfunction and injury caused by ischemia-reperfusion of the rat kidney. AB - This study was designed to determine the possible protective effect of mibefradil on renal ischemia/reperfusion (I/R) injury. Unilaterally nephrectomized Sprague Dawley rats were subjected to 60 min of left renal ischemia followed by 45 min of reperfusion. Group 1 were sham-operated animals; group 2, I/R/untreated animals; and group III, I/R/mibefradil-treated animals. A 99mTc-DTPA scan was taken to measure kidney perfusion, glomerular filtration rate (GFR) and the time elapsed from isotope injection to the maximum of the curve. Serum creatinine, blood urea nitrogen (BUN), kidney malondialdehyde (MDA) level were determined as well as examining the kidneys histologically. Treatment of rats with mibefradil produced a significant reduction in the serum levels of creatinine and urea nitrogen. T max-sec (renal perfusion) was significantly lower in group 2 than in groups 1 and 3. The GFR was markedly greater in group 3 than in the group 2. The Tmax-min was significantly greater in group 2 than in group 3. Mibefradil treatment significantly decreased the MDA levels. The histopathologic score was significantly less in the group 3 rats compared with group 2 rats. Kidneys of group 2 rats showed tubular cell swelling, cellular vacuolization, pyknotic nuclei, medullary congestion, and moderate to severe necrosis. Treatment with mibefradil preserved the normal morphology of the kidney and shows normal glomeruli and slight edema of the tubular cells. These findings suggest that mibefradil reduces the renal dysfunction associated with I/R of the kidney. PMID- 16350834 TI - Fabry disease--a diagnostic and therapeutic problem. AB - The authors present a patient with Fabry syndrome that remained undiagnosed for several years. Fabry syndrome is a genetic disease related to changes on the X chromosome. Its complex clinical presentation and diverse symptomatology is caused by deficient activity of lysosomal hydrolase alpha-galactosidase enzyme. Defect in the basic alpha-galactosidase molecule implies genetic change, which can be a predisposing factor for the development of atypical and typical forms of this genetic disease. In the presented case, clinical manifestation and hemizygous symptomatology were the evidence of metabolic and genetic irregularity, typical clinical presentation of Fabry disease. Many authors report generalized vasculopathy as a basic characteristic of Fabry disease and a causative factor of multiorgan changes. Some authors indicate that persons with diagnosed asymmetric hypertrophy of the left ventricle have decreased alpha galactosidase. Cardiac complications, coronary disease, and acute myocardial ischemia are often present in cases of Fabry disease, frequently causing death in such patients. Characteristic central nervous system symptoms with skin-burning sensation and paresthesia were also present in our case. Cerebrovascular complications were caused by changes on small blood vessels. Clinical signs of renal failure were nonspecific, and the diagnosis was based on extrarenal symptoms. Initial renal manifestations were insignificant as asymptomatic proteinuria and microhematuria, due to which our patient was referred to further examination. The level of alpha-galactosidase was significantly decreased. The severity and progression of this disease depends on the level of alpha galactosidase enzyme in serum and its catabolic effect. More recent studies have showed that deficient enzyme can be synthetic zed and, accordingly our patient has been successfully enrolled in the replacement therapy program. PMID- 16350835 TI - Acute renal failure after liposuction. PMID- 16350836 TI - Pulmonary renal syndrome due to anti-GBM and IgM C-ANCA disease requiring the use of novel therapeutic agents. AB - There are several known causes for the clinical syndrome of pulmonary hemorrhage and acute renal failure. Here, we report a unique case of a 50-year-old man presenting in this manner. The initial diagnosis was one of antiglomerular basement membrane (anti-GBM) disease that responded well to steroids, cyclophosphamide, and plasma exchange (PE). The pulmonary hemorrhage resolved, but he remained dialysis dependent. However, despite falling anti-GBM titers, the symptoms relapsed and standard therapy was reinitiated with limited success. The anti-GBM antibody titer fell to zero despite clinical deterioration, prompting a search for an alternative diagnosis. He was found to be IgM anti-proteinase-3 antineutrophil cytoplasmic antibody (C-ANCA) positive. The pulmonary hemorrhage responded successfully to the use of intravenous immunoglobulin and the antilymphocyte monoclonal antibody CD52. To our knowledge, this is the first known case of IgM C-ANCA in association with anti-GBM disease. As such, it highlights the predominance of pulmonary hemorrhage in this condition, as well as the need to consider alternative therapies in refractory cases. PMID- 16350837 TI - Unusual presentation of IgA nephropathy in childhood: a case report. AB - Immunoglobulin A (IgA) nephropathy is the most common form of primary glomerulonephritis worldwide, and approximately 20% to 30% of adult patients with the disorder develop chronic renal failure within 20 years. This type of nephropathy is also an important risk factor for chronic renal failure in children. The pathogenesis of IgA nephropathy is still unknown, and treatment remains controversial. Microscopic hematuria and recurrent episodes of macroscopic hematuria are the most common clinical manifestations of this condition in children. This article describes the case of a young girl who presented with steroid-resistant nephrotic syndrome unaccompanied by hematuria. Renal biopsy findings were consistent with IgA nephropathy. The patient's condition was a rare clinical manifestation of IgA nephropathy. PMID- 16350838 TI - Management of acute renal failure due to Russell's viper envenomation: an analysis on the reported Thai cases. AB - Russell's viper (RVV) is an important tropical venomous snake. In severe envenomous cases, acute renal failure (ARF) can be seen. Here, the author performs this ministudy to document the management of ARF among reported Thai patients. A literature review on the papers concerning ARF due to RVV in Thailand was performed. Based on this study, hemodialysis and specific antivenom administration shows more favorable outcome than peritoneal dialysis. PMID- 16350839 TI - Immune complex: does it have a role in pathogenesis of renal failure in dengue infection? AB - Dengue infection is a major public health problem. When acute renal failure complicates dengue infection, it is usually associated with severe disease as in dengue hemorrhagic fever or dengue shock syndrome. The role of immune complex in development of renal failure in dengue infection is still unclear. Here, the author used a computational medicine technology to study the property of the dengue virus-immunoglobulin complex. According to this study, the diameter of derived complex is much smaller, compared with the diameter of glomerulus. Entrapment of the immune complex is believed to occur when a previous glomerular lesion causes narrowing of the glomerulus's diameter. Therefore, the immune complex should not have a significant role in pathogenesis of renal failure in dengue infection. PMID- 16350840 TI - Effects of retinoic acid on the differentiation of chondrogenic progenitor cells, ATDC5. AB - Chondrocyte differentiation is a fundamental process during endochondral ossification. Retinoic acid (RA) has been shown to regulate this process, however, the mechanisms underlying RA regulation of chondrogenesis are not clearly understood. Chondroprogenitor cells, ATDC5 have been shown to be a useful in vitro model for examining the multiple step differentiation of chondrocytes. The present study investigated the mechanisms underlying RA regulation of chondrogenesis using ATDC5 cell culture. In this study, we show that RA suppresses the cell growth, cartilage nodule formation, accumulation of proteoglycan, alkaline phosphatase (ALPase) activity and mineralization and that RA dose dependently upregulates the levels of type X collagen and matrix metalloproteinase-13 (MMP-13) mRNA which are marker proteins of hypertrophic chondrocytes, in ATDC5 cells. The addition of protein synthesis inhibitor, cycloheximide (CHX), partially inhibits the induction of type X collagen and MMP 13 mRNA by RA. In this system, RA upregulates the mRNA level of Runx2/Cbfa1 (type II), a positive regulator for mineralization, and downregulates the mRNA of Indian hedgehog (Ihh), parathyroid hormone related protein (PTHrP), negative regulators for terminal differentiation. However, RA downregulates ALPase, bone gla protein (BGP) mRNAs and mineralization. These data indicate that RA stimulates cartilage differentiation, however, cell condensation and cartilage nodule formation may be candidates of primary importance in the terminal differentiation of chondrocytes. PMID- 16350841 TI - Clonality analysis for normal and cancerous colon tissues with human androgen receptor gene polymerase chain reaction. AB - We assessed the feasibility of clonality analysis with human androgen receptor gene polymerase chain reaction in terms of the sensitivity and specificity for normal and cancerous colonic tissues taken from fourteen informative cases selected from 22 women with colonic adenocarcinoma. Ten crypts microdissected from each 10-microm-thick cryostat sections and whole tissues were used as samples. DNA was extracted from the samples and amplified with and without prior enzyme digestion. These products were analyzed by capillary electrophoresis for clonality. Of the whole-tissue DNA, none of the normal tissues and seven (50.0%) of the cancerous tissues showed monoclonality. Of the microdissected samples, monoclonality was found in 88.4% (107/121) of normal crypts and 95.9% (117/122) of cancerous crypts. Samples composed of crypts with short and long alleles were found in eight of the 14 normal colonic mucosae, but in none of the cancerous tissues. We concluded that the sensitivity of this method is limited for both whole-tissue DNA and microdissected-tissue DNA, because monoclonality from small samples does not always indicate monoclonality of the entire lesion. The high specificity of this method, however, allows polyclonal results in whole tissues to be confirmed by additional analysis of microdissected tissues. PMID- 16350842 TI - Synthesis of new 2-thiouracil-5-sulphonamide derivatives with antibacterial and antifungal activity. AB - 2-Thiouracil-5-sulphonic acid N-(4-acetylphenyl) Amide (1) was reacted with a series of aromatic aldehydes giving chalcones 2 (Claisen-Schemidt reaction), some of these chalcones were reacted with urea and thiourea giving pyrimidine-2-one and pyrimidine-2 thione derivatives respectively of the type 3a,b and 4a,b. In addition many chalcones were reacted with hydroxylamine hydrochloride giving isoxazoline derivatives 5a,b. They could also reacted with phenylhydrazine to give pyrazoline derivatives 6a,b, chalcones also were reacted withethylcyano acetate and/or malononitryl in pyridine giving pyran derivatives 7a,c and 8a,c. In another pathway chalcones were epoxidised by H2O2 giving epoxides 9a,c which in turn were reacted with phenylhydrazine giving 4-hydroxypyrazoline derivatives 10a,c. In another reaction chalcones were reacted with ethylcyanoacetate in presence of amm.acetate giving pyridone derivatives 11a,d which could be prepared also in exellent yield from compound 1 by its reaction with certain aromatic aldehydes and ethylcyanoacetate in presence of ammonium acetate. Finally, compound 1 was reacted with semicarbazide giving semicarbazone intermediate 12 which in turn was reacted with thionyl chloride giving thiadiazole derivative 13. The biological effects of some of the new synthesized compounds were also investigated. PMID- 16350843 TI - Synthesis of new triazolyl-N,N-dialkyldithiocarbamates as antifungal agents. AB - N,N-Dialkylditihiocarbamate derivatives have been well known as broad-range fungicides. In this study, the triazole derivatives of ten new N,N-disubstituted dithiocarbamates (3a-j) were synthesized and their structures were identified by spectral and elemental analysis. Results of the antifungal activity studies showed that some of the compounds tested were active against M. canis, M. gypseum, and T. rubrum at the concentration of 12.5 microg/mL when clotrimazol was used as a standard. PMID- 16350844 TI - Synthesis of 6-[2-(benzoxazol-2-ylmethylamino)ethoxy]-1-alkyl-1H-indole-2 carboxylic acid and inhibitory activity on beta-amyloid aggregation. AB - 6-[2-(benzoxazol-2-ylmethylamino)ethoxy]-1-alkyl-1H-indole-2-carboxylic acids were designed and synthesized as beta-amyloid (Abeta) fibril assembly inhibitors. Their inhibitory activity on Abeta, aggregation was evaluated by thioflavin T assay although their activities were insignificant. PMID- 16350845 TI - Cytotoxic constituents from the whole plant of Corydalis pallida. AB - Here we report the cytotoxic activity of three known compounds isolated for the first time from Corydalis pallida (Papaveraceae). An isoquinoline alkaloid, berberine, exhibited cytototoxic activity against two human cancer cell lines, HT 1080 (human fibrosarcoma) and SNU-638 (human stomach adenocarcinoma), with IC50 values of 3.2 and 3.4 microg/mL, respectively. N-trans-feruloyltyramine and N trans-feruloylmethoxytyramine were also isolated from this plant but were inactive. PMID- 16350846 TI - Antibacterial and antitumor macrolides from Streptomyces sp. Is9131. AB - Four compounds, including two novel macrolides, were isolated from an endophyte Streptomyces sp. Is9131 of Maytenus hookeri. Spectral data indicated that these compounds were dimeric dinactin (1), dimeric nonactin (2), cyclo-homononactic acid (3), and cyclo-nonactic acid (4). Bioassay results showed that dimeric dinactin had strong antineoplastic activity and antibacterial activity. PMID- 16350847 TI - 3"-hydroxyamentoflavone and its 7-O-methyl ether, two new biflavonoids from Aristolochia contorta. AB - Two new biflavonoids, 3"-hydroxyamentoflavone-7-O-methyl ether (1) and 3" hydroxyamentoflavone (2), were isolated from the fruits of Aristolochia contorta Bge. Their structures were elucidated by HR-ESI-MS, 1D-, and 2D-NMR spectroscopy. PMID- 16350848 TI - Ophiopojaponin D, a new phenylpropanoid glycoside from Ophiopogon japonicus Ker Gawl. AB - A new phenolic glycoside, ophiopojaponin D (1), together with two known compounds, was isolated from the tubers of a famous traditional Chinese herb Ophiopogon japonicus Ker-Gawl. The spectroscopic and chemical data revealed their structures to be 3-tetradecyloxy-4-hydroxy-allylbenzene-4-O-alpha-L rhamnopyranosyl (1-->6)-beta-D-glucopyranoside (1), 3, 4-dihydroxy-allylbenzene-4 O-alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranoside (2) and L-pyroglutamic acid (3). PMID- 16350849 TI - Norditerpenoid alkaloids and other components from the processed tubers of Aconitum carmichaeli. AB - A new norditerpenoid and a known alkaloid were isolated from the alkaloidal fraction of the processed tubers of Aconitum carmichaeli. The structure of the new norditerpenoid alkaloid was elucidated as lipoforesaconitine (1) on the basis of spectroscopic analysis. The known norditerpenoid alkaloid was characterized as lipoyunanaconitine (2). In addition, a new flavonoid, 6"-O-acetylliquiritin (7), along with a known ceramide, (2S,3S,4R,8E)-2-[(2'R)-2'-hydroxylignoceroylamino] 8(E)-octadecene-1,3,4-triol (3), as well as a known steroid saponin, gracillin (8), and three known flavonoids, liquiritigenin (4), isoliquiritigenin (5), and liquiritin (6), were also isolated and characterized. All known compounds were isolated from this plant for the first time. The structures of the isolates were established by spectroscopic and chemical methods. PMID- 16350850 TI - Inhibitory effects of eutigosides isolated from Eurya emarginata on the inflammatory mediators in RAW264.7 cells. AB - The anti-inflammatory activity of Eurya emarginata (Thumb) Makino, of which leaves have been traditionally used to treat ulcers or diuretic in Jeju Island, has been investigated in the present study. Through the phytochemical study from the methanol extract of E. emaginiata, eutigosides B and C were isolated as the active components. Sseveral inflammatory markers including TNF-alpha, IL-1beta, IL-6, NO, iNOS, and COX-2 were examined. Eutigosides B and C potentially inhibited production of pro-inflammatory cytokines (IL-6 and TNF-alpha) in a dose dependent manner. Additionally, the intracellular contents of iNOS protein were markedly decreased after treatment with eutigosides B and C. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. These results suggest that eutigoside B and C from E. emarginata may have anti-inflammatory activity through the inhibition of pro-inflammatory cytokines (TNF-alpha and IL 6), iNOS and COX-2. PMID- 16350851 TI - Cytoprotective effect of green tea extract and quercetin against hydrogen peroxide-induced oxidative stress. AB - In this study, we evaluated the cytoprotective effects of antioxidative substances in hydrogen peroxide (H2O2) treated Mel-Ab melanocytes. Tested substances include selenium, quercetin, green tea (GT) extract, and several vitamins (ascorbic acid, Trolox, and folic acid). Of these, both quercetin and GT extract were found to have strong cytoprotective effects on H2O2-induced cell death. We also examined additive effects, but no combination of two of any of the above substances was found to act synergistically against oxidative damage in Mel Ab cells. Nevertheless, a multi-combination of GT extract, quercetin, and folic acid appeared to prevent cellular damage in a synergistic manner, which suggests that combinations of antioxidants may be of importance, and that co-treatment with antioxidants offers a possible means of treating vitiligo, which is known to be related to melanocyte oxidative stress. PMID- 16350852 TI - MAP kinase activation is required for the MMP-9 induction by TNF-stimulation. AB - MMP-9 is a metalloproteinase capable of basement membrane degradation in vivo. Expression of MMP-9 can be found in normal conditions such as trophoblasts, osteoclasts, and leukocytes and their precursors. They also occur as well as in pathological conditions, such as the invasive growth of primary tumors, metastasis, angiogenesis, rheumatoid arthritis, and periodontal diseases. MMP-9 upregulation can be highly induced by a wide range of agents. These agents include growth factors, cytokines, cell-cell, and cell-ECM adhesion molecules, and agents altering cell shape. Here, we observed that TNF-alpha stimulated human monocytic cell line, HL-60 produced MMP-9 in a dose and time dependent manner. Real time PCR results indicated transcriptional upregulation of MMP-9 as early as 3 h post TNF-alpha stimulation. To investigate the signaling pathway underlined in TNF-alpha induced MMP-9 expression, three MAP kinase inhibitors were added to cells 1 h prior to TNF-alpha treatment. The ERK inhibitor completely abolished MMP-9 expression by TNF-alpha. But neither p38 MAP kinase nor JNK inhibitor had an effect on TNF-alpha induced MMP-9 expression, suggesting that ERK activation is required for the MMP-9 induction by TNF-alpha. Taken together, we found that TNF-alpha stimulation facilitates ERK activation, which results in the transcriptional upregulation of MMP-9 gene and subsequent MMP-9 production and secretion. PMID- 16350853 TI - Adiponectin induces growth arrest and apoptosis of MDA-MB-231 breast cancer cell. AB - Recently, it was reported that reduction in serum adiponectin levels is correlated with the incidence of breast cancer. As an effort to explain this, we screened various human breast cancer cell lines to identify those in which proliferation is directly controlled by adiponectin. Among the five tested cell lines, proliferation of MDA-MB-231 cancer cell was significantly suppressed by adiponectin within the range of physiological concentration. Furthermore, prolonged adiponectin treatment caused cell growth arrest and even apoptosis of MDA-MB-231. This result is the first to show that adiponectin can directly control cancer cell growth and provides a rationale for the theory that reduction in plasma adiponectin levels could be a risk factor for breast cancer. PMID- 16350854 TI - Sanguiin H-6 blocks endothelial cell growth through inhibition of VEGF binding to VEGF receptor. AB - The vascular endothelial growth factor (VEGF) plays a key role in angiogenesis, which is a process where new blood vessels develop from the endothelium of a pre existing vasculature. VEGF exerts its activity by binding to its receptor tyrosine kinase, KDR/Flk-1, which is expressed on the surface of endothelial cells. A methanol extract and organic solvent (n-hexane, ethyl acetate, n butanol, aqueous) fractions from Rubus coreanus were examined for their inhibitory effects on VEGF binding to the VEGF receptor. The methanol extract from the crude drug were found to significantly inhibit VEGF binding to the VEGF receptor (IC50 approximately = 27 microg/mL). Among the fractions examined, the aqueous fraction from the medicinal plant showed potent inhibitory effects against the binding of KDR/Flk-1-Fc to immobilized VEGF165 in a dose-dependent manner (IC50 approximately = 11 microg/mL). Sanguiin H-6 was isolated as an active principle from the aqueous fraction, and inhibited the binding of KDR/Flk 1-Fc to immobilized VEGF165 in a dose-dependent manner (IC50 approximately = 0.3 microg/mL). In addition, sanguiin H-6 efficiently blocked the VEGF-induced HUVEC proliferation in a dose-dependent manner (IC50 approximately = 7.4 microg/mL) but had no effect on the growth of HT1080 human fibrosarcoma cells. This suggests that sanguiin H-6 might be a potential anti-angiogenic agent. PMID- 16350855 TI - Direct involvement of G protein alpha(q/11) subunit in regulation of muscarinic receptor-mediated sAPPalpha release. AB - The G(q/11) protein-coupled receptors, such as muscarinic (M1 & M3) receptors, have been shown to regulate the release of a soluble amyloid precursor protein (sAPPalpha) produced from alpha-secretase processing. However, there is no direct evidence for the precise characteristics of G proteins, and the signaling mechanism for the regulation of G(q/11) protein-coupled receptor-mediated sAPPalpha release is not clearly understood. This study examined whether the muscarinic receptor-mediated release of sAPPalpha is directly regulated by Galpha(q/11) proteins. The HEK293 cells were transiently cotransfected with muscarinic M3 receptors and a dominant-negative minigene construct of the G protein alpha subunit. The sAPPalpha release in the media was measured using an antibody specific for sAPP. The sAPPalpha release enhancement induced by muscarinic receptor stimulation was decreased by a G(q/11) minigene construct, whereas it was not blocked by a control minigene construct (the Galpha carboxy peptide in random order, Galpha(q)R) or Galpha(i) constructs. This indicated a direct role of the Galpha(q/11) protein in the regulation of muscarinic M3 receptor-mediated sAPPalpha release. We also investigated whether the transactivation of the epidermal growth factor receptor (EGFR) by a muscarinic agonist could regulate the sAPPalpha release in SH-SY5Y cells. Pretreatment of a specific EGFR kinase inhibitor, tyrophostin AG1478 (250 nM), blocked the EGF stimulated sAPPalpha release, but did not block the oxoM-stimulated sAPPalpha release. This demonstrated that the transactivation of the EGFR by muscarinic receptor activation was not involved in the muscarinic receptor-mediated sAPPalpha release. PMID- 16350856 TI - Immunomodulatory function of murine NK cell activity by alginate. AB - The in vivo immunomodulatory function of the activity of murine natural killer (NK) cells induced by high mannuronic acid-containing alginate (HMA) was examined. HMA was injected i.p at doses of 25 and 100 mg/kg. The NK activity was 3 times higher with 100 mg/kg HMA than the baseline. In addition, in vitro studies of splenocytes cultured with HMA for 20 h showed a significant increase in NK activity at E:T ratio of 100:1; a 160% and 210% increase at 10 and 100 microg/mL, respectively. There was a six fold increase in interferon-gamma production in a postculture of splenocytes with 100 microg/mL HMA. HMA had no suppressive effects on the lymphocyte function in the presence or absence of mitogens. This suggests that HMA is useful in cancer immunotherapy. PMID- 16350857 TI - In vitro metabolism of a new cardioprotective agent, KR-33028 in the human liver microsomes and cryopreserved human hepatocytes. AB - KR-33028 (N-[4-cyano-benzo[b]thiophene-2-carbonyl]guanidine) is a new cardioprotective agent for preventing ischemia-reperfusion injury. This study was performed to identify the metabolic pathway of KR-33028 in human liver microsomes and to compare its metabolism with that of cryopreserved human hepatocytes. Human liver microsomal incubation of KR-33028 in the presence of NADPH and UDPGA resulted in the formation of four metabolites, M1, M2, M3, and M4. M1 and M2 were identified as 5-hydroxy-KR-33028 and 7-hydroxy-KR-33028, respectively, on the basis of LC/MS/MS analysis with the synthesized authentic standard. M3 and M4 were suggested to be dihydroxy-KR-33028 and hydroxy-KR-33028-glucuronide, respectively. Metabolism of KR-33028 in cryopreserved human hepatocytes resulted in the formation of M1, M2, and M4. These data show a good correlation between major metabolites formed in human liver microsomes and cryopreserved human hepatocytes. In addition, KR-33028 was found to inhibit moderately the metabolism of CYP1A2 substrates. Based on the results obtained metabolic pathway of KR-33028 is proposed. PMID- 16350858 TI - Antiherpetic activities of flavonoids against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in vitro. AB - Flavonoids, a group of low molecular weight phenylbenzopyrones, have various pharmacological properties including antioxidant, anticancer, bactericidal, and anti-inflammatory. We carried out anti-herpetic assays on 18 flavonoids in five classes and a virus-induced cytopathic effect (CPE) inhibitory assay, plaque reduction assay, and yield reduction assay were performed. When flavonoids were applied at various concentrations to Vero cells infected by HSV-1 and 2, most of the flavonoids showed inhibitory effects on virus-induced CPE. Among the flavonoids, EC, ECG (flavanols), genistein (isoflavone), naringenin (flavanone), and quercetin (flavonol) showed a high level of CPE inhibitory activity. The antiviral activity of flavonoids were also examined by a plaque reduction assay. EC, ECG, galangin, and kaempferol showed a strong antiviral activity, and catechin, EGC, EGCG, naringenin, chrysin, baicalin, fisetin, myricetin, quercetin, and genistein showed moderate inhibitory effects against HSV-1. In these experiments, flavanols and flavonols appeared to be more active than flavones. Furthermore, treatment of Vero cells with ECG and galangin (which previously showed strong antiviral activities) before virus adsorption led to a slight enhancement of inhibition as determined by a yield reduction assay, indicating that an intracellular effect may also be involved. PMID- 16350859 TI - Glucosylated polyethylenimine as a tumor-targeting gene carrier. AB - Glucosylated polyethylenimine (GPEI) was synthesized as a tumor-targeting gene carrier through facilitative glucose metabolism by tumor glucose transporter. Particle sizes of GPEI/DNA complex increased in proportion to glucose content of GPEI, whereas surface charge of the complex was not dependent on glucosylation, partially due to inefficient shielding of the short hydrophilic group introduced. GPEI with higher glucosylation (36 mol-%) had no cytotoxic effect on cells even at polymer concentrations higher than 200 microg/mL. Compared to unglucosylated PEI, glucosylation induced less than one-order decrease of transfection efficiency. Transfection of GPEI/DNA complex into tumor cells possibly occurred through specific interaction between glucose-related cell receptors and glucose moiety of GPEI. Gamma imaging technique revealed GPEI/DNA complex was distributed in liver, spleen, and tumors. PMID- 16350860 TI - Enhancing dermal matrix regeneration and biomechanical properties of 2nd degree burn wounds by EGF-impregnated collagen sponge dressing. AB - To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 microg/1.7 cm2. Dermal fibroblasts were cultured to 80% confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, 2nd degree- burn of diameter 1cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 +/- 35.6 vs. 411.5 +/- 57.0 Fmax, gmm(-2)) and skin resilience (11.3 +/- 1.4 vs. 6.5 +/- 0.6 mJ/mm2) were significantly increased with EGF-treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, 2nd degree-burns enhanced the dermal matrix formation. PMID- 16350861 TI - Fortune favours the prepared mind: a national perspective on pandemic preparedness. AB - The rapid spread of avian influenza H5N1 in Asia in late 2003 has moved the world to a new state of pandemic alert. The risk of further human cases will persist, as will opportunities for a pandemic virus to emerge. Therefore, Canada has taken a number of steps to strengthen national pandemic preparedness. Coordination has been fostered through the development of a national pandemic preparedness plan and ongoing development of systems and processes for national health emergency management. Specific areas of advancement include: enhancement of rapid surveillance and communication capacity, the pandemic vaccine strategy, acquisition of an antiviral stockpile, research prioritization, international collaboration, and an international meeting of Ministers of Health (October 2005) to enhance global cooperation and coordination in advance of a possible influenza pandemic. Key next steps include: publication of the 2005 edition of the Canadian Pandemic Influenza Plan; conducting emergency exercises to help strengthen planning at all levels and across health care, emergency management and NGO sectors; developing H5N1 vaccine and clinical trials; evaluating the need to supplement national antiviral stockpile; and undertaking public and key stakeholder consultations to provide further input into planning strategies and activities. PMID- 16350862 TI - Ensuring a broad and inclusive approach: a provincial perspective on pandemic preparedness. AB - The SARS crisis revealed critical gaps in Ontario's health emergency response capacity, and identified, in the starkest terms possible, the need for improved emergency response planning. This article reviews the development of the Ontario Health Plan for an Influenza Pandemic (OHPIP), released in June 2005. Some key points arising from the provincial planning process include the necessity to: ensure a broad and inclusive development process; ensure the pandemic plan identifies: 1) clear roles and responsibilities of federal, provincial/territorial and municipal levels of government, 2) the approach to occupational health and safety issues and ethical decision-making, 3) a communications strategy linking all affected sectors and levels of government and health sector; 4) any commitments to antiviral stockpiling, vaccine and antiviral allocation and use, and an approach for drug delivery from provincial stockpiles to local public health units; 5) health human resource management and supplementation; and 6) key programs/services to be scaled back to maximize surge capacity; address best practices (e.g., involve all sectors of the health care system at the outset, acquire strategic expertise, coordinate/advocate with broader emergency response system, etc); and, outline future stages that include strengthening the delivery of clinical care to influenza cases; clarifying the role of primary care practitioners during a pandemic; leveraging Ontario's significant e-Health investments. PMID- 16350863 TI - Taking care of the sick and scared: a local response in pandemic preparedness. AB - Virtually all health care operations, including public health, are undertaken only at a local or regional level. Large-scale infectious disease emergencies, such as SARS or pandemic influenza, will be recognized and managed at a local level. The creation of the Public Health Agency of Canada (PHAC) was an important step in strengthening public health capacity. However, we need adequate operational capacity in local public health departments to have a strong public health system. Local public health takes an integral role in the preparation for and management of infectious disease emergencies. Local public health departments and regional public health infrastructures must be positioned to both maintain core functions and to lead and support health sector response to emergencies. The local establishment of a flexible and sustainable emergency management system must address the need to: integrate health care and first responders; provide all hazards tools for managing a crisis at the frontline; rank service priorities and provide surge resources; and provide accurate information on a timely basis. Only the leaders within the local or regional health care facilities and organizations can develop workable plans to deliver health care. PHAC must ensure and support the local public health infrastructure and local emergency preparedness. Without this support, there will be consequences for local response to major public health emergencies. PMID- 16350864 TI - Neighbourhood socio-economic status and the prevalence of overweight Canadian children and youth. AB - BACKGROUND: The purposes of this study are to determine (i) if neighbourhood socioeconomic status (SES) is systematically related to the prevalence of overweight children and youth in Canada, (ii) if the factors accounting for the apparent relationship have face validity, and (iii) if neighbourhood SES has an independent influence on this distribution. METHODS: Cross-sectional data from Cycle 4 (2000/2001) of the National Longitudinal Survey of Children and Youth were used. Children and youth aged 5 to 17 were included. Overweight was established using age and sex cut-off points. Neighbourhood socioeconomic data were obtained from the Statistics Canada 2001 Dissemination Area databases and SES quartiles constructed using a composite of socio-economic variables. Hierarchical non-linear modelling was used to test for independent neighbourhood effects. RESULTS: A gradient of increasing overweight prevalence by decreasing neighbourhood SES quartiles was observed (24% high SES, 30% mid-high SES, 33% mid low SES, 35% low SES). Controlling for individual age, gender, family income and education hierarchical analysis found that a child's odds of being overweight increases if living in a low versus a high SES neighbourhood (OR=1.29, 95% CI=1.14-1.46). INTERPRETATION: The prevalence of child and youth overweight in Canada is inversely and statistically significantly related to neighbourhood SES. Independent effects indicate that neighbourhood characteristics directly influence the odds of being overweight. This research suggests that consideration of opportunity structures that exist in different types of neighbourhoods is fundamentally important to health promotion and disease prevention strategies. PMID- 16350865 TI - Promotion of healthy weights at preschool public health vaccination clinics in Calgary: an obesity surveillance program. AB - OBJECTIVE: To evaluate the acceptability and feasibility of a surveillance program of overweight and obesity in preschool children in Calgary, and to provide advice for families to promote healthy weights. PARTICIPANTS AND SETTING: Children (mean age 4.9+/-0.6 years) attending pilot-site clinics in September 2002 and all clinics in Calgary between February 2003-December 2003 (n=7048). INTERVENTION: The growth assessment protocol and resources supported a three pronged approach to promote healthy weights (healthy eating, active living and positive body image). Public health nurses were trained in standardized measurement techniques and information resources. Links with physicians were made to facilitate continuity of care. Children's weight and height measurements were plotted on the Weight-for-Stature growth chart and used to identify children as obese (> or =95th percentile), healthy weight (> or =5th, <95th) or underweight (<5th). Subsequent analysis calculated the Body Mass Index (BMI)-for-Age to identify overweight children (> or =85th, <95th percentile). The protocol was pilot tested and subsequently implemented in all Calgary public health clinics. OUTCOMES: The majority (98%) of parents were either very happy or happy with information received during the visit. Public health nurse counselling confidence significantly improved after the pilot (p<0.001). Data indicated that 9% of children were obese, 15% were overweight and 3% were underweight. INTERPRETATION: This approach to identifying children's weight status appeared satisfactory to stakeholders, maximized use of existing resources to establish a surveillance program for Calgary, and provided an opportunity to give parents health-promoting advice on healthy weights. PMID- 16350866 TI - Investigating health correlates of adolescent depression in Canada. AB - BACKGROUND: The prevalence of depression rises sharply during adolescence. Understanding health correlates of adolescent depression may provide descriptive information with regard to which adolescents are more likely to be depressed. Health determinants have been found to have associations with depression in adult populations, but have never been investigated concurrently with depression in a national sample of adolescents in Canada. Therefore, the aim of the present investigation was to understand which health determinants would be significantly associated with adolescent depression. METHODS: A sample of 17,557 adolescents was used from the Canadian Community Health Survey (CCHS) 1.1 to determine the health correlates of adolescent depression among males and females. To understand the relationship between health determinants, logistic regressions were conducted. RESULTS: The survey had an 84.7% response rate. The past 12-month prevalence of depression among the sample of adolescents was 6.5% +/- 0.4% (3.4% +/- 0.27% for males and 9.8% +/- 0.44% for females). Reporting fair/poor perceived health, smoking, alcohol dependence, food allergies, migraine headaches, chronic bronchitis, and having physical health conditions had positive associations with depression for males and females. However, gender differences in the relationship between health correlates and depression were found. Even after controlling for all variables, females were still more likely to be depressed. CONCLUSIONS: Several health determinants were associated with depression in adolescents in Canada. However, the relationship between some health determinants and depression functioned differently for males and females. PMID- 16350867 TI - The health of street youth: a Canadian perspective. AB - OBJECTIVE: To review epidemiologic studies of the health of street youth in industrialized countries, with a special focus on Canadian youth. METHODS: We identified 52 peer-reviewed studies from searches of the MEDLINE database and bibliographies of published papers, for data on blood-borne and sexually transmitted infections, mental health problems, pregnancy, violence and mortality. RESULTS: Rates of hepatitis B, hepatitis C, and HIV infection are much higher among street youth than among their non-street peers. Likewise, the prevalence of all mental health problems assessed in street youth is greater than that in non-street youth. Pregnancy is more frequent among street than household youth. Street youth also experience high levels of violence: a large proportion report physical abuse or assault. Finally, mortality is about 11 times the expected rate based on age and sex and is mainly caused by suicide and drug overdose. CONCLUSION: Current research results are useful to orient public health interventions for street youth, but further epidemiologic research is needed. The need for Canadian data is particularly acute in specific areas including mental health, violence, pregnancy, and sexually transmitted infections such as, for example, herpes infection and syphilis. PMID- 16350868 TI - [Use of condoms by adolescents seeking oral contraceptives in the Montreal area]. AB - OBJECTIVES: To examine female adolescents' use of condoms before and after seeking their first prescription of oral contraceptives and to identify variables associated with use of oral contraceptives. METHOD: This longitudinal study was conducted among 350 female adolescents recruited from 8 CLSCs in the Montreal Metropolitan Area. Data were collected at the first medical visit and again three months later. RESULTS: At the 3-month follow-up, a decrease of 41% was observed in the systematic use of condoms with an inadequate use of condoms reported in 44% of subjects. In the last year, the variables associated with irregular use of condoms were low intention to use simultaneously the pill and the condom in the next 12 months, higher frequency of intercourse, a larger number of sexual partners, and alcohol or drug use before or during sexual intercourse. CONCLUSION: Our results indicate that female adolescents stop using condoms when they adopt oral contraceptives. At the first visit, the lack or inadequate use of condoms seemed related to other STD risk behaviours. These results should encourage clinicians to revise their counselling approach with adolescents and promote dual protection in the context of serially monogamous relationships with known partners or with partners they trust. PMID- 16350869 TI - Are national vitamin D guidelines sufficient to maintain adequate blood levels in children? AB - BACKGROUND: Vitamin D insufficiency (defined as 25-hydroxyvitamin D [25(OH)D] concentrations <40 nmol/L) may be associated with subclinical adverse effects on bone mineralization. The current vitamin D status of children and adolescents in Canada has not been described. The purpose of this study was to describe the association between 25(OH)D serum concentration and dietary vitamin D intake, and other potential determinants of vitamin D status, among a sample of children and adolescents aged 2-16 years presenting to a pediatric emergency department in Edmonton, Alberta (latitude 52 degrees N) at the end of winter. METHODS: In early April 2003, 90 patients between the ages of 2 and 16 years who presented to the pediatric emergency department in Edmonton volunteered to participate. All participants and/or parents or guardians completed questionnaires regarding potential risk factors for vitamin D insufficiency, detailed dietary assessments, and anthropometric measurements. Serum 25(OH)D concentrations were measured in 68 of 90 participants. RESULTS: The mean serum 25(OH)D concentration was 47.2 nmol/L (95% CI 43.8-50.8 nmol/L). 34% of participants had vitamin D insufficiency (<40 nmol/L) and 6% were deficient (<25 nmol/L). Boys and girls aged 9-16 years had a prevalence of insufficiency of 69% and 35% respectively, while boys and girls 2-8 years old had a prevalence of insufficiency of 22% and 8% respectively. Dietary vitamin D intake per kilogram body weight was the most important independent determinant of 25(OH)D concentration (r = 0.446, p<0.001). Vitamin D intake, age and male sex best predicted insufficiency. No subject was insufficient if they had an intake >0.45 mcg/kg/day. INTERPRETATION: Vitamin D insufficiency may be common among children and adolescents at the beginning of spring. The risk may be highest among older children because vitamin D intake does not adequately rise in proportion with increases in body mass. Further studies are needed to assess whether Canadian dietary vitamin D recommendations should be changed. PMID- 16350870 TI - Framework for population health. PMID- 16350871 TI - [Crack smokers and harm reduction]. PMID- 16350872 TI - Registered indians and tobacco taxation: a culturally-appropriate strategy? AB - Taxation of tobacco is a widely-used strategy that prompts smoking cessation among adults and reduces cigarette consumption among continuing smokers. Registered Indian tobacco use prevalence is at least double that of the rest of Canadians and is in part due to the lower cost of tobacco products purchased on reserve by Registered Indians (RIs) as they are tax exempt. Although registered Indian communities have the ability to collect tax on tobacco products and direct the use of these revenues, this strategy is rarely utilized. Tobacco taxation could have substantial health and economic benefits to RI communities, but perhaps is not culturally-appropriate. In order to better support RI communities, governments and other organizations need to examine this policy instrument in the context of RI populations. PMID- 16350873 TI - Access to drugs for cancer: Does where you live matter? AB - BACKGROUND: Provincial governments are responsible for administering publicly funded anti-cancer drug benefit programs in Canada. This study examines inter provincial variations in not only the content of such programs, but also the policies/processes used when considering a new drug for coverage. METHODS: Pharmaceutical manufacturers and provincial/regional cancer boards were surveyed to identify the drugs covered by public drug benefit plans. Kappa coefficients were calculated to determine inter-provincial coverage variations. The comprehensiveness of availability of anti-cancer drugs across the country was also assessed. A semi-structured survey of all 10 provincial/regional cancer board pharmacy and therapeutics (P&T) committees was employed to examine decision making policies/procedures. It included questions on committee composition and processes and on factors influencing decisions regarding the introduction of new drugs. Completed surveys were analyzed using qualitative and quantitative techniques. RESULTS: All cancer boards and 75% of manufacturers contacted provided information on drugs covered in each province. Where lists were obtained from both sources, there was full agreement on content. Kappa values calculated ranged from -0.403 to 0.594, indicating poor to moderate agreement on anti-cancer drug coverage between provinces. Only 7 of the 115 drugs were available in all 10 provinces. Regarding decision-making processes, while ratings for both the relative importance and use of factors involved in decision-making (clinical effectiveness, patient preference, etc.) were similar across provinces, those for the relative importance and use of different information types (clinical trials, expert opinion, etc.) varied. CONCLUSION: Access to anti-cancer drugs clearly varies across the country. In part, this may be due to differences in the views of P&T committees on the usefulness of information they use in their deliberations. PMID- 16350874 TI - Prescription drug abuse in Canada and the diversion of prescription drugs into the illicit drug market. AB - Prescription drug abuse has received considerable attention in media reports in recent years. The purpose of this article is to describe the Canadian situation and context with regards to prescription drug abuse and the diversion of psychotropic prescription drugs into the illicit drug market, with a focus on the need for more data and interventions. Canada ranks within the top 10% of countries in the use of benzodiazepines, opioid prescriptions and stimulants. There are many ways that prescription drugs are diverted into the illicit market and varied reasons for use and abuse. Prescription drug abuse is further related to a number of negative consequences, including overdose. While seniors and women have been the primary focus for research in Canada on prescription drug abuse, adolescents and young adults have received less attention. Systematic epidemiological data specifically on prescription drug abuse in the Canada context are lacking and are needed in order to more clearly understand the reasons for the phenomenon and to develop and implement appropriate interventions. PMID- 16350875 TI - Compared to whom? An investigation of the relative health comparisons of well people. AB - OBJECTIVES: Responses to the self-reported health (SRH) question, "In general how would you rate your health? Excellent, very good, good, fair or poor", have been repeatedly demonstrated to predict mortality. Survival curves plotted for each response category show that the likelihood of death increases in a stepped fashion with each progressively negative response category and the relationship persists for up to 12 years following initial measurement. To whom do people compare themselves when answering the self-reported health question? METHODS: Twenty-one interviews with people who selected the better-health response choices (i.e., good, very good, or excellent) were conducted in a Canadian city. Qualitative content analysis was undertaken and the interviews were approached with no pre-conceived ideas about what the participants would say. RESULTS: Analysis of the interviews suggested that there are three key features in making a health comparison: the type (i.e., with whom), direction (i.e., upward or downward), and magnitude of the comparison (i.e., with a few people or an ideal person vs. many other people). These key features varied in a systematic way with the chosen response. DISCUSSION: The findings related to the direction of comparison contradict, somewhat, the theories of social psychologists but these differences may have occurred because the participants were well. Healthy people may use different factors than ill people when making social comparisons as they select a self-reported health question response category. The nature of the comparisons are complex and the responses indicate that they are affected by age, gender, and life experience. PMID- 16350876 TI - Health promotion and disease prevention for older adults: Intervention themes and strategies used in Quebec local community health centres and seniors' day centres. AB - OBJECTIVE: Despite the considerable potential of disease prevention and health promotion (DPHP) among older adults, extant data suggest that this field of intervention is still underdeveloped. To shed further light on this issue, this paper presents the results of an inventory of DPHP interventions for older adults conducted in local community health centres (CLSCs) and seniors' day centres in the province of Quebec. METHODS: All CLSCs (N=147) and day centres (N=124) were invited to participate (response rates: 74% and 79%). Data were collected through telephone interviews. Interventions were coded according to type of intervention strategies and target themes. RESULTS: Awareness-raising and health education strategies emerged as the most frequently-cited type of intervention strategies, reported by 77% of CLSCs and 95% of day centres, respectively. The two themes reported by a majority of CLSCs were physical health (87%) and community issues (58%). Lifestyle habits (92%) and social issues (92%) were the two most frequently-cited themes in day centres. DISCUSSION: DPHP for older adults is substantially well developed in terms of intervention offerings in the two types of organizations under study. However, the range of available interventions requires expansion to increase the potential of DPHP programs to tackle the numerous challenges posed by the aging of the population. PMID- 16350877 TI - Sporadic infections of Salmonella Paratyphi B, var. Java associated with fish tanks. AB - OBJECTIVE: Identify a source of contamination to explain the increase in the number of cases of S. Paratyphi B var. Java infection. METHOD: A descriptive study was conducted to gather more information about cases and potential risk factors for infection. Between January 2000 and June 2003, we identified and interviewed by questionnaire 53 people with S. Paratyphi B infection. Due to the suspected link with fish tanks, an environmental investigation was conducted for each case at patients' homes as well as at pet stores and wholesalers. Various samples were taken from fish tanks. The fish were purchased at different pet shops. The pulsed field gel electrophoresis method was used to compare the human and aquarium strains. RESULTS: Of those infected, 60% had bought or had contact with an aquarium or tropical fishes before becoming ill. More than 50% of the samples taken from fish tanks in homes, pet shops, and wholesaler operations contained a number of serotypes of Salmonella in addition to Salmonella Paratyphi B var. Java. There were similarities between the PFGE patterns of human strains and aquarium samples. DISCUSSION: Contact with fish tanks could be a risk factor for developing a Salmonella infection and the public may be unaware of this risk. It was recommended that information be produced and distributed to pet shops and their customers to inform them of safety precautions in order to reduce the risk of contamination from bacterial pathogens that may be present in aquarium water. PMID- 16350878 TI - The tip of the iceberg. PMID- 16350879 TI - Fluoridation and public health. PMID- 16350880 TI - Will oral health assessment bill make the grade? PMID- 16350881 TI - California's dental schools have changed. PMID- 16350882 TI - Issues facing dental education and how to address them. PMID- 16350883 TI - Dental faculty recruitment at Loma Linda University School of Dentistry. AB - This article addresses the dental faculty shortage as reported by the American Dental Association in its publication The Future of Dentistry and the Journal of Dental Education report on the demand nationally for dental school faculty. Budgeted full-time faculty vacancies in U.S. dental schools stand between 300 and 400 with present and anticipated state and federal shortfalls forecasting even further cuts or at least lack of government support. Acknowledging various reasons for, and responses to, the shortage, the School of Dentistry at Loma Linda University is seeking to deal creatively with a faculty shortage. A survey of newly hired full-time faculty indicates the issues that concern them and their colleagues. Loma Linda University School of Dentistry's response to the shortage includes a routine invitation to students, particularly seniors and residents, to consider teaching at the school. In addition, a simple alumni recruitment form is used for personal relationship building with alumni. A student loan reimbursement program for new faculty is a successful program in recruiting new, young full time faculty. To provide for the long-term future of the research function of the school, a new program focusing on hiring doctoral (PhD)-prepared individuals who will be trained in the predoctoral DDS program at the expense of the school is in place. This article suggests further steps that can be taken to enhance the cause and reputation of the school's educational program. PMID- 16350884 TI - How does a dental school ensure the currency of practitioners who graduated some years ago. PMID- 16350885 TI - Changing financial base for dental education. AB - In recent years, there have been material changes in the manner in which U.S. dental schools are funded. This fact has had profound ramifications for the practices, operations, and culture of the schools. The kinds of programs supported, patients treated, and students admitted all are affected by the sources of available revenue as schools can only do that which does not jeopardize their economic viability. PMID- 16350886 TI - Educating the practice-ready dentist. AB - The mission of the University of the Pacific, Arthur A. Dugoni School of Dentistry is to graduate competent beginning dentists in a humanistic environment. As the first American dental school to incorporate competency-based education, it takes very seriously the understanding that competency means having the skills, understanding, and supporting values to begin independent practice. Competency is defined by the demands of dental practice, not by what professors know. The university's mission statement is to "provide a superior, student centered learning experience integrating liberal arts and professional education, and prepare individuals for lasting achievement and responsible leadership in their careers and communities". PMID- 16350887 TI - Emerging scientific advances: how do they enter dental curricula and the profession? AB - What is meant by emerging scientific advances? In brief, this terminology is equivalent to new research findings, however, the term "research" is often associated with scientific investigations that have very limited direct clinical relevance. Unfortunately, basic dental research and dental clinical practice have, in many instances, been considered to have nonoverlapping spheres of existence. The existence of mutually exclusive domains is rapidly changing with considerable translational activities between basic research investigation and clinical application developing. There is a growing emphasis at a national level for the importance of moving basic biomedical research laboratory findings into clinical patient-related applications to realize improvements in health based on the research findings. Ultimately, new approaches to diagnose, treat, and prevent disease will be available and represent the translation of the best scientific evidence into clinical applications. It is critical at this time to prepare our dental graduates to be members of the dental profession who will understand the implications that new scientific advances will have on their approach to patient care. The patterns and practices of oral health care delivery will undergo major changes during the careers of our new dental graduates. They need to be prepared to respond to these changes to the benefit of their patients. PMID- 16350888 TI - Can we talk? PMID- 16350889 TI - Evaluation of enzyme inhibitors in drug discovery. A guide for medicinal chemists and pharmacologists. PMID- 16350891 TI - Developing new peer reviewers: tips for editors. PMID- 16350890 TI - How accurate are references in nursing journals? PMID- 16350892 TI - After Katrina. PMID- 16350893 TI - Will we ever answer Katrina's wake-up call? PMID- 16350894 TI - The high cost of being a moral chameleon. PMID- 16350895 TI - Market program boosts staff retention. PMID- 16350896 TI - Toward a healthy work environment. AB - Patient safety and the quality of patient care have become important topics in U.S. hospitals in recent years. Nearly all hospitals have made mechanical and procedural changes to improve safety. But a truly healthy hospital is one in which caregivers, especially nurses, are treated with respect. The American Association of Critical-Care Nurses argues that a genuinely healthy work environment has six qualities: skilled communication, true collaboration, effective decision making, appropriate staffing, meaningful recognition of staff members' contributions, and authentic leadership. Although training is often needed to create a respectful and healthy workplace, some leaders are reluctant to allocate dollars on what they see as "soft" relationship issues. However, hospital leaders dedicated to improving safety and quality of care will see that they cannot afford to neglect such issues. PMID- 16350897 TI - Breaking the turnover cycle. AB - The Paraprofessional Healthcare Institute, Bronx, NY, identifies seven elements which are essential for employers in their creation of a stable, well-qualified direct-care workforce. These "common sense" elements--ranging from paying family sustaining wages to strengthening the caregiving relationship between the workers and those whom they serve--can make the difference between chronic turnover, on one hand, and employing a dedicated staff large enough to keep pace with the growing need for long-term care and services, on the other. However, implementing the seven elements requires the commitment, coordination, and the cooperation of the entire long-term care system, especially the state and federal reimbursement systems that pay for most care and services. By joining forces with other long term care stakeholders, through organizations such as the Direct Care Alliance, providers can help stabilize the workforce and, ultimately, help to increase the allocation of public resources. PMID- 16350898 TI - Turnover turnaround. AB - The Foundation for Long Term Care (FLTC) created a program called Growing Strong Roots to help long-term care providers retain certified nurse aides (CNAs) and to enhance the quality of care that new CNAs provide. To adapt elements of existing peer-mentoring programs from around the country, Growing Strong Roots revised them and added new components, including administrator training, supervisor training, "booster sessions," newsletters, and a multisite evaluation. The program focuses on helping new CNAs become part of a facility's culture. Growing Strong Roots was evaluated in 11 diverse nursing homes. Research shows that nursing homes using the system increased their new CNA retention rates by 25 percent. A control group showed an increase of only 10 percent. All nursing homes participating in the project improved their retention rates, some by as much as 41 percent. In March 2005, the American Society on Aging, in collaboration with the Brookdale Center on Aging of Hunter College, New York City, honored Growing Strong Roots with a Best Practices in Human Resources and Aging Award. PMID- 16350899 TI - The ethics of recruiting foreign nurses. AB - Because they face a growing nursing shortage, many U.S. health care institutions have turned to recruiting foreign nurses. For foreign nurses, the practice is often an opportunity to make a better life for themselves and their families. And it helps solve a serious problem for the U.S. organizations involved. But the recruitment of foreign nurses raises a number of ethical questions. The first article here examines the practice as seen from three viewpoints, the global, that of the particular recruiting health care organization, and that of the recruited foreign nurse. The author concludes that the practice can be both a "blessing" and a "curse." The second article discusses the practice as seen from a Third World nation from which the United States, along with other Western countries, is recruiting nurses. The author, who formerly supported the practice, now opposes it. PMID- 16350900 TI - India is losing its nurses to the West. PMID- 16350901 TI - Who will care for the caregivers? AB - Research shows that stable leadership provided by a nursing home's director of nursing (DON) is the foundation of good quality. Conversely, a high DON turnover often results in a quality meltdown at all levels. A recent study, seeking to uncover the root causes of DON turnover, looked at 103 DONs working in Virginia nursing homes. Although DONs overwhelming affirmed their role and agreed that they were overall satisfied, a majority said they would not choose to be a DON again or recommend the role to others. Not surprisingly, low staff retention rates, mounting administrative responsibilities, growing regulatory and legal constraints, and unrealistic time commitments were the leading causes of frustration. Given less time to spend on the responsibilities that lie at the very heart of their mission-direct-care and clinical issues, quality improvement, family relations, staff mentoring and professional development-DONS are likely to experience burnout unless long-term care owners and managers reconfigure their role. PMID- 16350902 TI - Magnetic direction. AB - In April 2005, the American Nurses Association (ANA) awarded St. Joseph Hospital, Nashua, NH, its highest honor for excellence in nursing: "Magnet Recognition." The Magnet Recognition Program was developed by the ANA's American Nurses Credentialing Center in the early 1980s to recognize health care organizations that provide the best in nursing care and uphold the tradition of excellence in professional nursing practice. St. Joseph began pursuing Magnet status more than three years ago, starting with a number of enhancements to nursing practices. The hospital worked hard to improve nurse-to-patient staffing and included many of its nurses on the nursing quality council, division advisory, and cultural diversity committees. Magnet program appraisers visited the hospital this January to conduct an intensive, on-site three-day examination. They interviewed patients, staff nurses, physicians, hospital employees, administrators, board members, and nursing leadership to evaluate St. Joseph's nursing care, services, and delivery of care to patients and their families. Soon after, Magnet status was bestowed. PMID- 16350903 TI - Canon law is flexible and adjustable. PMID- 16350904 TI - Is the NovaSure system ethical? PMID- 16350905 TI - Training spiritual care volunteers. PMID- 16350906 TI - Knowledge and action: developing cultural competence. PMID- 16350907 TI - Helping family members manage medication administration hassles. AB - Managing the medication regimens of dependent older adults in the home is a complex issue for family caregivers. The tasks involved can include helping the older adults manage their own medications or may require involvement in the purchasing, scheduling, and administering of multiple medications for serious conditions. The hassles associated with this responsibility are now known to be associated with increased caregiver strain. In this article, we provide an overview of the body of research on medication administration hassles and offer a tool to assess family caregivers who are involved in this important aspect of care. PMID- 16350908 TI - "To no longer be a mystery to myself": living with PTSD. PMID- 16350909 TI - Working with inmates: educational preparation. PMID- 16350910 TI - "The right stuff" is right on. PMID- 16350911 TI - Disaster response for mental health professionals. Interview by Shirley A. Smoyak. PMID- 16350912 TI - Screening for depression and suicide: self-report instruments that work. AB - Self-report instruments for depression and suicide risk can help busy clinicians identify adult clients who may be at risk for depressive disorders. These instruments provide a baseline measure of clients' behavioral status, reflect response to treatment, and improve clinical decision making. Such scales can also detect symptoms of depression regardless of whether they are reported or observed. Clinicians' recognition of depression is improved when self-report screening tools are used. When it is not feasible to conduct a thorough psychiatric evaluation, screening instruments can identify at-risk clients who need further evaluation. PMID- 16350913 TI - Implementing a smoking ban in an acute psychiatric admissions unit. AB - In contrast to general medical hospitals, psychiatric hospitals often allow patients to smoke cigarettes. In addition to obvious health concerns, smoking can also interfere with clinical assessments and therapeutic activities, Implementation of a smoking ban on an acute male admissions unit did not result in any increase in aggressive behaviors. In addition, staff attitudes following the ban improved, and most staff members believed the ban was both ethical and beneficial to patients. Our research indicates that banning smoking on an acute admissions unit is feasible and well tolerated by patients and staff, although it may require extra vigilance for smoking-related contraband. PMID- 16350914 TI - Community need for university-based mental health services: identifying the top 5. AB - The purpose of this study was to determine the community need for mental health and health promotion services provided by advanced practice nursing faculty at a small university in the midwestern United States. Fifty participants from a three county service area were selected to complete a needs assessment survey. Survey results indicated strong community need for primary mental health services for low-income residents, older adults, pregnant women, and families with children. Mental health services (particularly for depression), health promotion, wellness information, and primary mental health care were among the leading areas of need. In addition, low-cost mental health services were identified as the most significant need, which was magnified in the underserved, Hispanic population. PMID- 16350915 TI - Are we really helping? The problem of dual diagnoses, homelessness, & hospital hopping. PMID- 16350916 TI - Comparison and validation of tissue modelization and statistical classification methods in T1-weighted MR brain images. AB - This paper presents a validation study on statistical nonsupervised brain tissue classification techniques in magnetic resonance (MR) images. Several image models assuming different hypotheses regarding the intensity distribution model, the spatial model and the number of classes are assessed. The methods are tested on simulated data for which the classification ground truth is known. Different noise and intensity nonuniformities are added to simulate real imaging conditions. No enhancement of the image quality is considered either before or during the classification process. This way, the accuracy of the methods and their robustness against image artifacts are tested. Classification is also performed on real data where a quantitative validation compares the methods' results with an estimated ground truth from manual segmentations by experts. Validity of the various classification methods in the labeling of the image as well as in the tissue volume is estimated with different local and global measures. Results demonstrate that methods relying on both intensity and spatial information are more robust to noise and field inhomogeneities. We also demonstrate that partial volume is not perfectly modeled, even though methods that account for mixture classes outperform methods that only consider pure Gaussian classes. Finally, we show that simulated data results can also be extended to real data. PMID- 16350917 TI - Restrictions on the three-class ideal observer's decision boundary lines. AB - We are attempting to develop expressions for the coordinates of points on the three-class ideal observer's receiver operating characteristic (ROC) hypersurface as functions of the set of decision criteria used by the ideal observer. This is considerably more difficult than in the two-class classification task, because the conditional probabilities in question are not simply related to the cumulative distribution functions of the decision variables, and because the slopes and intercepts of the decision boundary lines are not independent; given the locations of two of the lines, the location of the third will be constrained depending on the other two. In this paper, we attempt to characterize those constraining relationships among the three-class ideal observer's decision boundary lines. As a result, we show that the relationship between the decision criteria and the misclassification probabilities is not one-to-one, as it is for the two-class ideal observer. PMID- 16350918 TI - Virtual angiography for visualization and validation of computational models of aneurysm hemodynamics. AB - It has recently become possible to simulate aneurysmal blood flow dynamics in a patient-specific manner via the coupling of three-dimensional (3-D) X-ray angiography and cmputational fluid dynamics (CFD). Before such image-based CFD models can be used in a predictive capacity, however, it must be shown that they indeed reproduce the in vivo hemodynamic environment. Motivated by the fact that there are currently no techniques for adequately measuring complex blood velocity fields in vivo, in this paper we describe how cine X-ray angiograms may be simulated for the purpose of indirectly validating patient-sperific CFD models. Mimicking the radiological procedure, a virtual angiogram is constructed by first simulating the time-varying injection of contrast agent into a precomputed, patient-specific CFD model. A time-series of images is then constructed by simulating the attenuation of X-rays through the computed 3-D contrast-agent flow dynamics. Virtual angiographic images and residence time maps, here derived from an image-based CFD model of a giant aneurysm, are shown to be in excellent agreement wiith the corresponding clinical images and residence time maps, but only when the interaction between the quasisteady contrast agent injection and the pulsatile flow are properly accounted for. These virtual angiographic techniques pave the way for validating image-based CFD models against routinely available clinical data, and provide a means of visualizing complex, 3-D blood flow dynamics in a clinically relevant manner. They also clearly show how the contrast agent injection perturbs the noraml blood flow patterns, further highlighting the potential utility of image-based CFD as a window into the true aneurysmal hemodynamics. PMID- 16350919 TI - Maximum-likelihood techniques for joint segmentation-classification of multispectral chromosome images. AB - Traditional chromosome imaging has been limited to grayscale images, but recently a 5-fluorophore combinatorial labeling technique (M-FISH) was developed wherein each class of chromosomes binds with a different combination of fluorophores. This results in a multispectral image, where each class of chromosomes has distinct spectral components. In this paper, we develop new methods for automatic chromosome identification by exploiting the multispectral information in M-FISH chromosome images and by jointly performing chromosome segmentation and classification. We (1) develop a maximum-likelihood hypothesis test that uses multispectral information, together with conventional criteria, to select the best segmentation possibility; (2) use this likelihood function to combine chromosome segmentation and classification into a robust chromosome identification system; and (3) show that the proposed likelihood function can also be used as a reliable indicator of errors in segmentation, errors in classification, and chromosome anomalies, which can be indicators of radiation damage, cancer, and a wide variety of inherited diseases. We show that the proposed multispectral joint segmentation-classification method outperforms past grayscale segmentation methods when decomposing touching chromosomes. We also show that it outperforms past M-FISH classification techniques that do not use segmentation information. PMID- 16350920 TI - Automated detection of prostatic adenocarcinoma from high-resolution ex vivo MRI. AB - Prostatic adenocarcinoma is the most commonly occurring cancer among men in the United States, second only to skin cancer. Currently, the only definitive method to ascertain the presence of prostatic cancer is by trans-rectal ultrasound (TRUS) directed biopsy. Owing to the poor image quality of ultrasound, the accuracy of TRUS is only 20%-25%. High-resolution magnetic resonance imaging (MRI) has been shown to have a higher accuracy of prostate cancer detection compared to ultrasound. Consequently, several researchers have been exploring the use of high resolution MRI in performing prostate biopsies. Visual detection of prostate cancer, however, continues to be difficult owing to its apparent lack of shape, and the fact that several malignant and benign structures have overlapping intensity and texture characteristics. In this paper, we present a fully automated computer-aided detection (CAD) system for detecting prostatic adenocarcinoma from 4 Tesla ex vivo magnetic resonance (MR) imagery of the prostate. After the acquired MR images have been corrected for background inhomogeneity and nonstandardness, novel three-dimensional (3-D) texture features are extracted from the 3-D MRI scene. A Bayesian classifier then assigns each image voxel a "likelihood" of malignancy for each feature independently. The "likelihood" images generated in this fashion are then combined using an optimally weighted feature combination scheme. Quantitative evaluation was performed by comparing the CAD results with the manually ascertained ground truth for the tumor on the MRI. The tumor labels on the MR slices were determined manually by an expert by visually registering the MR slices with the corresponding regions on the histology slices. We evaluated our CAD system on a total of 33 two-dimensional (2-D) MR slices from five different 3-D MR prostate studies. Five slices from two different glands were used for training. Our feature combination scheme was found to outperform the individual texture features, and also other popularly used feature combination methods, including AdaBoost, ensemble averaging, and majority voting. Further, in several instances our CAD system performed better than the experts in terms of accuracy, the expert segmentations being determined solely from visual inspection of the MRI data. In addition, the intrasystem variability (changes in CAD accuracy with changes in values of system parameters) was significantly lower than the corresponding intraobserver and interobserver variability. CAD performance was found to be very similar for different training sets. Future work will focus on extending the methodology to guide high-resolution MRI-assisted in vivo prostate biopsies. PMID- 16350921 TI - Delay correlation subspace decomposition algorithm and its application in fMRI. AB - This paper reports a new delay subspace decomposition (DSD) algorithm. Instead of using the canonical zero-delay correlation matrix, the new DSD algorithm introduces a delay into the correlation matrix of the subspace decomposition to suppress noises in the data. The algorithm is applied to functional magnetic resonance imaging (fMRI) to detect the regions of focal activities in the brain. The efficiency is evaluated by comparing with independent component analysis and principal component analysis method of fMRI. PMID- 16350922 TI - Retributive justice and dentistry. PMID- 16350923 TI - Entrepreneurship in continuing dental education: a dental school perspective. AB - The definition of continuing dental education is presented, along with its benefits to the profession. The preeminence of dental schools in providing lifelong learning opportunities and freedom from commercial involvement that existed even twenty years ago has changed. Less than a quarter of CE takes place in school, and the focus there is increasingly on material with deep scientific background and hands-on learning. The newest innovations and those with the greatest commercial potential are taught elsewhere. Proposed changes in the ADA CERP standards would take on a "purist" approach that could place dental schools at a severe disadvantage while allowing "for profit" institutes to flourish and thus further undermine the role dental schools can play in providing quality professional development experiences. PMID- 16350924 TI - Industry support for dental education. AB - The relationships between industry and dental education are multiple and mutually beneficial. Perhaps most prominent are collaboration on research and development of products and technologies and the knowledge and public credibility that accompany them. Industry is also looked to for product and equipment support in schools and increasingly for help with outreach and access programs schools provide for underserved populations. Not as widely recognized, but still quite important, are the programs for support of student research and sharing of management expertise through exchange of board members. A quarter century ago, the relationship between schools and industry was at arm's length. There was a mistrust in schools that feared exposing their students to commercial contact. Today the relationship has evolved into a mutual search for joint benefits with an eye on the future of the profession and its relationship with patients. This is illustrated in the American Dental Education Association Corporate Council. PMID- 16350925 TI - The buy or lease decision in dental curricula. AB - Dental schools are aware of the need to adequately prepare graduates for the business aspects of dentistry in addition to the clinical, biomedical science, and interpersonal skills now necessary to succeed in practice. The challenge includes the alternatives of building such a program with existing faculty members (buy) or bringing in the kind of management program that practicing dentists would select (lease). Marquette University School of Dentistry has had three successful years of using the Pride Institute on a contract basis to provide a comprehensive practice management program. PMID- 16350926 TI - Entrepreneurialism in dentistry: commercialism versus opportunity. AB - The school of dentistry at Nova Southeastern University was founded eight years ago at a time when other dental schools were closing. Efficiencies of structure with the Health Professions Division of the university have been important in making this possible. The applicant pool is large and diverse and the educational program is modern. Collaborative relationships are being developed with industry. PMID- 16350927 TI - Founders of a profession: the original subscribers to the first dental journal in the world. AB - A true profession is built upon a tripod: a formal organization, formal professional education, and a formal scientific literature. The United States was the leader in all three. In 1839-40, the American Society of Dental Surgeons was organized, the Baltimore College of Dental Surgery was established, and the first dental journal in the world, the American Journal of Dental Science, was founded. At that time there were only about three hundred trained and scientific dentists in the entire country; the rest were relatively untrained operators, outright quacks, or charlatans. In 1898, a list of the first subscribers to the first journal was discovered and published by G.V. Black. These initial subscribers may be considered the core group of truly professional American dentists. They became the leaders of the newly born profession of dentistry. Short biographies of some of them are included. PMID- 16350928 TI - Errors in dentistry: a call for apology. AB - Bad outcomes occur in dentistry and sometimes these are the results of dental errors. In both cases, this essay will argue that apologies are very important in maintaining a relationship with the patient that is based on trust and mutual respect. Nevertheless, apologies are often not forthcoming in dentistry for a number of reasons that deserve careful examination. In particular, the dentist's fear that an apology will increase the risk of legal harm will be critiqued. Ethical and psychological reasons for making an apology will be discussed, and strategies to assist clinicians in making an apology will be offered. PMID- 16350929 TI - The Progressive Era. AB - The American College of Dentists was founded in 1920 for the purpose of encouraging young dentists to continue study and to apply science to their practices. This ideal emerged in the Progressive Era, which lasted roughly from 1895 to 1920. The animating spirit of this period was that the human condition could be improved and that the way to achieve this was through science and the use of experts working together. The Progressive Era saw inventions, such as automobiles and airplanes, telephone and radio, that required mass production and brought people together. It also spawned many political and legislative innovations that we now take for granted. Among these are the Food and Drug Administration, the Department of Commerce, and the Federal Trade Commission. Workers' compensation and other social protections were introduced, as were city commissions; the income tax; women's suffrage; and initiative, referendum, and recall. Medicine, for the first time, became an effective way to treat disease as it developed a scientific foundation. PMID- 16350930 TI - Global status of mumps immunization and surveillance. PMID- 16350931 TI - Being "normal". PMID- 16350932 TI - The Gaols that spawn disease. PMID- 16350933 TI - Gender-specific aspects of cardiovascular disease. PMID- 16350934 TI - Irritable bowel syndrome. PMID- 16350935 TI - Update in surgical management of female stress urinary incontinence. PMID- 16350936 TI - Obesity in women. PMID- 16350937 TI - A 20 cm mass in a 28-year-old woman. PMID- 16350938 TI - Future of breast surgery. PMID- 16350939 TI - Antidepressant-induced weight gain. PMID- 16350940 TI - Overweight and weight control among Rhode Island girls and women, 2003. PMID- 16350941 TI - Rhode island department of health office of women's health: a gender focus to improve health status. PMID- 16350942 TI - Are older Rhode Island women receiving appropriate screening for breast cancer? PMID- 16350943 TI - Introduction to chemical genomics. PMID- 16350944 TI - Chemistry for chemical genomics. AB - New methods and strategies have been developed to design and use small molecules that allow the functional dissection of molecular pathways, cells, and organisms by selective small-molecule ligands or modulators. In this overview, we are focusing on diversity aspects, design methods, and chemical synthesis strategies for the application of small molecules as tools for chemical genomics. Examples for different successful chemical-genomics strategies include the selection of diverse drug-like molecules, target family-focused compound libraries, natural product chemistry, and diversity-oriented synthesis. PMID- 16350945 TI - Computer-aided design of small molecules for chemical genomics. AB - De novo design provides an in silico toolkit for the design of novel molecular structures to a set of specified structural constraints, and is thus ideally suited for creating molecules for chemical genomics. The design process involves manipulation of the input, modification of structural constraints, and further processing of the de novo-generated molecules using various modular toolkits. The development of a theoretical framework for each of these stages will provide novel practical solutions to the problem of creating compounds with maximal chemical diversity. This chapter describes the fundamental problems encountered in the application of novel chemical design technologies to chemical genomics by means of a formal representation. Formal representations help to outline and clarify ideas and hypotheses that can then be explored using mathematical algorithms. It is only by developing this rigorous foundation, that in silico design can progress in a rational way. PMID- 16350946 TI - Design, synthesis, and screening of biomimetic ligands for affinity chromatography. AB - Affinity chromatography is ideally suited to the purification of pharmaceutical proteins due to its unique bio-specificity characteristics. Tailor-made affinity ligands that represent a promising class of synthetic affinity ligands have been developed to target specific proteins and designed to mimic peptidal templates, natural biological recognition motifs, or complementary surface-exposed residues. These biomimetic ligands have been generated by a combination of rational design, combinatorial library synthesis, and subsequent screening of potential leads against target proteins. Small ligands based on a triazine scaffold also present exceptional selectivity and stability, which allows their use in harsh manufacturing environments. PMID- 16350947 TI - The role and application of in silico docking in chemical genomics research. AB - In silico docking techniques are being used to investigate the complementarity at the molecular level of a ligand and a protein target. As such, docking studies can be used to identify the structural features that are important for binding and for in silico screening efforts in which suitable binding partners can be identified. Here we describe a practical approach for setting up docking simulations using different docking programs. We also cover the analysis and rescoring of the obtained docking poses. Possible pitfalls in the docking studies are discussed and hints are provided to resolve commonly occurring problems. PMID- 16350948 TI - Synthesis of complex carbohydrates and glyconjugates: enzymatic synthesis of globotetraose using alpha-1,3-N-acetylgalactosaminyltransferase LgtD from Haemophilus infuenzae strain Rd. AB - The lipopolysaccharide of capsule-deficient Haemophilus infuenzae strain Rd contains an N-acetylgalactosamine residue attached to the terminal globotriose moiety in the Hex5 glycoform. Genome analysis identified an open reading frame, HI1578, referred to as LgtD, whose amino acid sequence shows a significant level of similarity to those of a number of bacterial glycosyltransferases involved in lipopolysaccharide biosynthesis. To investigate its function, overexpression and biochemical characterization were performed. Most of the protein was obtained in a highly soluble and active form. Standard glycosyltransferase assay, high performance liquid chromatography (HPLC), and liquid chromatography (LC)/mass spectrometry (MS) show that LgtD is an N-acetylgalactosaminyltransferase with high donor substrate specificity, and globotriose is a highly preferred acceptor substrate for the enzyme. PMID- 16350949 TI - High-throughput cloning for proteomics research. AB - Ligation-independent cloning (LIC) is a simple, rapid, and efficient method for high-throughput cloning. In this system, linear plasmid vector and insert DNA are treated to generate complementary single-stranded overhangs that anneal during a short incubation. The LIC system is adaptable for use with any vector following an alteration of the vector sequence. This chapter describes the creation of an LIC-compatible vector, with tips on how to make any vector LIC-enabled. It also includes a protocol for generating high-quality linearized vector template for the LIC reaction. Lastly, a step-by-step protocol of the LIC reaction is outlined, with useful tips and tricks for optimization and screening. PMID- 16350950 TI - Screening for the expression of soluble recombinant protein in Escherichia coli. AB - Protein expression and purification have traditionally been time-consuming, case specific endeavors, and are considered to be the greatest bottlenecks in most proteomics pipelines. Escherichia coli (E. coli) is the most convenient and cost effective host, although optimal conditions for the expression of different proteins vary widely. Proteins vary in their structural stability, solubility, and toxicity in this environment, resulting in differing rates of protein degradation, formation into insoluble inclusion bodies, and cell death, thus affecting the amount of soluble protein that can be obtained from E. coli grown in culture. To take full advantage of a variety of strategies developed to improve the expression of soluble protein in E. coli, an easy, rapid means to test many growth parameters is necessary. This chapter describes a dot-blot expression screen to test the effects of growth and induction parameters on the yield of soluble protein. The expression screen is used to detect hexahistidine tagged proteins expressed in E. coli; however, it is adaptable for the detection of other affinity tags or fusion partners that have suitable antibodies available. In this example, induction time and temperature are tested; however, it can be used to test additional parameters, such as affinity tag type and placement, E. coli host type, and growth medium formulations. PMID- 16350951 TI - High-throughput purification of hexahistidine-tagged proteins expressed in E. coli. AB - This chapter describes a method for efficient high-throughput purification of hexahistidine-tagged proteins that are expressed in Escherichia coli (E. coli) using immobilized metal affinity chromatography (IMAC) in a 96-well format. This approach is particularly suitable for proteomic applications that require modest amounts of highly purified proteins to be generated very efficiently. This approach is also very useful for identifying protein targets that are most amenable to scaled-up production for use in structural studies. The typical yield of proteins purified using this system is 50-150 microg, which is generally greater than that of many in vitro expression systems and much less costly. The method as described has been optimized for purifying approx 150 microg of hexahistidine-tagged protein, but the method is flexible, so that the amount of affinity matrix and culture volumes can be adjusted for optimal binding capacity and consequently highest purity. Although the method detailed here uses IMAC to purify hexahistidine-tagged proteins, this basic platform can be used with many other tags and affinity resins. PMID- 16350952 TI - The wheat germ cell-free expression system: methods for high-throughput materialization of genetic information. AB - This chapter contains protocols for high-throughput protein production based on the cell-free system prepared from eukaryote wheat embryos. PMID- 16350953 TI - Advances in genome-wide protein expression using the wheat germ cell-free system. AB - In the current post-genomic era, cell-free translation platforms are gaining importance in structural as well as functional genomics. They are based on extracts prepared from Escherichia coli cells, wheat germ, or rabbit reticulocytes, and when programmed with any mRNA in the presence of energy sources and amino acids, can synthesize the respective protein in vitro. Among the cell-free systems, the wheat germ-based translation system is of special interest due to its eukaryotic nature and robustness. This chapter outlines the existing protein production platforms and their limitations, and describes the basic concept of the wheat germ-based cell-free system. It also demonstrates how the conventional wheat germ system can be improved by eliminating endogenous inhibitors, by using an expression vector specially designed for this system and polymerase chain reaction-directed protein synthesis directly from cDNAs in a bi layer translation system. Finally, a robotic procedure for translation based on the wheat germ extract and bi-layer cell-free translation is described. PMID- 16350954 TI - Production of proteins for NMR studies using the wheat germ cell-free system. AB - This chapter describes protocols for preparing 15N-labeled proteins (ubiquitin is used as an example) using Escherichia coli cells (with purification) and the wheat germ cell-free system (without purification). A comparison of 1H-15N heteronuclear single-quantum coherence (HSQC) spectra of yeast ubiquitin prepared using each method indicates that this wheat germ cell-free system may be used for rapid nuclear magnetic resonance analyses of proteins without purification. PMID- 16350955 TI - Fabrication of protein function microarrays for systems-oriented proteomic analysis. AB - Protein microarrays have many potential applications in high-throughput analysis of protein function. However, simple, reproducible, and robust methods for array fabrication are required. Here we discuss the background to different routes to array fabrication and describe in detail one approach in which the purification and immobilization procedures are combined into a single step, dramatically simplifying the array fabrication process. We illustrate this approach by reference to the creation of an array of p53 variants, and discuss methods for assay and data analysis on such arrays. PMID- 16350956 TI - Peptide mass fingerprinting: protein identification using MALDI-TOF mass spectrometry. AB - Matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF)-mass spectrometry (MS) is now routinely used in many laboratories for the rapid and sensitive identification of proteins by peptide mass fingerprinting (PMF). We describe a simple protocol that can be performed in a standard biochemistry laboratory, whereby proteins separated by one- or two-dimensional gel electrophoresis can be identified at femtomole levels. The procedure involves excision of the spot or band from the gel, washing and de-staining, reduction and alkylation, in-gel trypsin digestion, MALDI-TOF MS of the tryptic peptides, and database searching of the PMF data. Up to 96 protein samples can easily be manually processed at one time by this method. PMID- 16350957 TI - A practical protocol for carbohydrate microarrays. AB - We have established a high-throughput biochip platform for constructing carbohydrate microarrays. Using this technology, carbohydrate-containing macromolecules of diverse structures, including polysaccharides, natural glycoconjugates, and mono- and oligosaccharides coupled to carrier molecules, can be stably immobilized on a glass chip without chemical modification. Here, we describe a practical protocol for this technology. We hope that anyone who has access to a standard cDNA microarray facility will be able to explore this technology for his or her own research interest. We also provide an example to illustrate that the carbohydrate microarray is also a discovery tool; this is particularly useful for identifying immunologic sugar moieties, including complex carbohydrates of cancer cells and sugar signatures of previously unrecognized microbial pathogens. PMID- 16350959 TI - Diagnostic and prognostic guidelines for the vegetative and minimally conscious states. AB - Many individuals who sustain severe brain injury experience prolonged or permanent disorders of consciousness. While these disorders may appear homogeneous, important distinctions exist in prognosis and clinical management. Studies suggest, however, that the incidence of diagnostic inaccuracy is high in both acute care and rehabilitation settings. In this paper, we review consensus based diagnostic and prognostic criteria for the vegetative and minimally conscious states. We also discuss recent developments and future directions for research in this area. PMID- 16350960 TI - The vegetative state and the Royal College of Physicians guidance. AB - The Royal College of Physicians of the UK, together with the Colleges of Edinburgh and Glasgow, have produced guidance on the diagnosis and management of people in the vegetative state (report of a working party of the Royal College of Physicians, 2003). Such guidance is important when the single criterion for awareness in an individual is the perception of that awareness by a potentially fallible observer. The current guidance is reviewed and comparisons made with existing arrangements in other countries. Consideration is given to the possibility of future improvements in diagnosis with the advent of imaging and metabolic assessments of brain function and the need to define the required qualifications and training for those "experts" who are currently involved in the diagnosis of the vegetative state. PMID- 16350961 TI - Incidence and prevalence of the vegetative and minimally conscious states. AB - The methodological difficulties of obtaining accurate epidemiological data for vegetative state (VS) and minimally conscious state (MCS) are considered, and prompt the conclusion that published data are of uncertain validity, partly due to variation in the criteria for diagnosis. On the basis of these data, incidence of VS continuing for at least six months arises at a rate of between 5 and 25 per million population (PMP). The prevalence of VS in adults in the US is between 40 and 168 PMP, and may be lower in the UK, but precise figures are not available. The incidence and prevalence of MCS have yet to be established. PMID- 16350962 TI - Recovery of consciousness and life expectancy of children in a vegetative state. AB - The vegetative state does occur in children and is most commonly due to acquired traumatic and non-traumatic injuries. However, neurometabolic and degenerative diseases, as well as certain developmental brain malformations such as anencephaly, can also cause this condition. There are limited data available in children concerning recovery of consciousness and function from the vegetative state as well as life expectancy. This review concentrates on these issues and is based primarily on the data published in the Multi-Society Task Force Report on PVS which was published in 1994 as well as other epidemiological studies. Children in a vegetative state do have a poor prognosis for recovery of consciousness and function and do have a shortened life expectancy. Further research is needed to better understand what variables might contribute to recovery and what therapies might be of benefit. PMID- 16350963 TI - Neuropathology of the vegetative state after head injury. AB - A detailed neuropathological study of patients identified clinically after head injury as either severely disabled (SD, n = 30) or vegetative (VS, n = 35) has been carried out to determine the nature and frequency of the various pathologies that form the basis of these clinical states. Patients who were SD were older (SD median 49.5 yrs vs. VS median 38 yrs, p = .04), more likely to have a lucid interval (SD 31% vs. VS 9%, p = .03), and to have had an acute intracranial haematoma (SD 70% vs. VS 26%, p < .001). SD patients less often had severe, Grades (2 or 3) of traumatic diffuse axonal injury (SD 30% vs. VS 71%, p = .001) and less often had thalamic damage (SD 37% vs. VS 80%, p < .001). Similar features of both focal and diffuse damage were present in some SD and VS cases with both groups having considerable damage to white matter and to the thalamus. It is concluded that the principal structural basis of both SD and VS is diffuse traumatic axonal injury (DAI) with widespread damage to white matter and changes in the thalami. However, both ischaemic brain damage and the vascular complications of raised intracranial pressure contributed to the clinical signs and symptoms. PMID- 16350964 TI - The neurological assessment of patients in vegetative and minimally conscious states. AB - Patients with serious central nervous system (CNS) injuries can enter a chronic state in which there is impaired awareness, but the presence of preserved basic vegetative functions. Such cases present formidable challenges to the medical practitioner in terms of clinical assessment and thus management. This paper presents the neurological approach to such patients highlighting the clinical clues that need to be sought to decide whether the patient is in a persistent vegetative or minimally conscious state (i.e., showing any responsiveness to external stimuli), or alternatively suffering from a locked-in syndrome. This neurological clinical formulation is then assimilated with other tests and assessments from a range of medical specialities, and by so doing helps confirm the clinical impression. This multidisciplinary approach is vital in the assessment of such patients and ideally should also include neurophysiological and functional imaging paradigms. PMID- 16350965 TI - Hydrocephalus, ventriculomegaly and the vegetative state: a review. AB - The revised guidelines from the Royal College of Physicians make it clear that structural problems such as hydrocephalus should be excluded before the diagnosis of vegetative state (VS) can be made. Ventriculomegaly is common after severe head injury but the distinction between atrophy and potentially treatable hydrocephalus cannot be made on the basis of conventional computerised tomographic (CT) or magnetic resonance (MR) scanning alone--physiological measurements of intracranial pressure (ICP) and cerebrospinal fluid (CSF) outflow resistance may be helpful. These techniques are reviewed together with the limited literature available that documents the effect of CSF diversion on outcome in "vegetative" patients. PMID- 16350966 TI - Permanent vegetative state: the legal position. AB - Medicine's diagnostic and therapeutic capacities raise increasingly complex ethical and legal issues for consideration. This is particularly so when the patient is in a permanent vegetative state. This article reviews the current legal position in the case of adults in permanent vegetative state (pVS), with particular attention to the devices used by courts in reaching decisions about whether or not to prolong assisted nutrition and hydration. The article further considers the impact of the Human Rights Act 1998, and argues that the current legal position is significantly, if not determinatively, based on clinical judgement, even where there is some doubt about whether or not the cases actually meet the terms of the Royal College of Physicians (RCP) diagnostic guidelines. In addition, the article asks whether or not the devices used by courts to permit withdrawing assisted nutrition and hydration from patients in pVS could be categorised as assisted death, and notes the (arguable) weakness of these devices as a basis for derogation from the sanctity of life principle. PMID- 16350967 TI - Forensic assessment issues in low level neurological states. AB - Forensic assessment of persons in low-level neurological state (LLNS) is fraught with limitations and caveats. Examiners must be aware of the nuances of forensic evaluation in the context of the conditions of examination, ethics and responsibilities that they have been charged with as an independent evaluator and fact seeker in the context of litigation-oriented dispute resolution. This article provides an overview of the independent medical evaluation (IME) of persons in LLNS including ethics, contextual limitations, history gathering, corroboratory interviews and suggested examination, as well as report format, among other important issues. PMID- 16350968 TI - Ethics and the vegetative state. AB - Before discussing ethical issues to do with patients in permanent (or persistent) vegetative state (PVS) it is necessary to address the foundational issue of whether PVS as a concept is able to provide a robust link to situations in the real world. The high reported rates of misdiagnosis and recovery in patients diagnosed as being in PVS casts doubt upon the applicability of ethicists' thought experiments on Platonic forms to actual decision making in clinical situations. We should abandon the illusion that we can have access to logical certainty through diagnostic definition, and should instead frame our opinions and our procedures in ways that can accommodate a high element of uncertainty, and should in the light of recent studies give considerable weight to the possibility that patients, at present unable to express opinions on their care, will later become able to do so, if given proper treatment and adequate evaluation. PMID- 16350969 TI - The vegetative state: promoting greater clarity and improved treatment. AB - The condition commonly referred to as the persistent vegetative state (PVS) or vegetative state (VS) generates tremendous confusion among health care professionals. Muddled and nihilistic views of very severe brain injury have hampered efforts to improve the diagnosis and treatment of patients thought to be in the VS. Significant obstacles to diagnostic clarity arise from multiple sources including imprecise terminology and conflation of the concepts of "behaviour" and "awareness". Failure to employ effective, uniform protocols of assessment and rehabilitation contributes to inadequate treatment of these extremely vulnerable patients. Despite diagnostic and prognostic difficulties, courts across the globe have accepted medical opinion as persuasive evidence for life-support withdrawal. A new outlook on severe brain injury is needed, with greater clarity and standardisation of assessment and care. Best practices in assessment and rehabilitation must be incorporated along with new developments in cognitive neuroscience and neuroimaging. Such a rehabilitative view will encourage intellectual curiosity towards improved quality of care for patients with severe brain injury. Attaining high levels of accuracy depends upon reaching a clearer understanding of the nature of human consciousness itself, of the condition, and of the patient's potential for full or partial recovery. PMID- 16350970 TI - Part II: Functional imaging, electrophysiology and mechanical intervention. PMID- 16350971 TI - Functional imaging of vegetative state applying single photon emission tomography and positron emission tomography. AB - Nuclear medicine techniques, such as single photon emission tomography (SPECT) and positron emission tomography (PET) have been applied in patients in a vegetative state to investigate brain function in a non-invasive manner. Parameters investigated include glucose metabolism, perfusion at rest, variations of regional perfusion after stimulation, and benzodiazepine receptor density. Compared to controls, patients in a vegetative state show a substantial reduction of glucose metabolism and perfusion. While patients post-anoxia exhibit a rather homogenous cortical reduction of glucose metabolism, patients after head trauma often show severe cortical and sub-cortical reductions at the site of primary trauma. To distinguish reduced glucose metabolism due to neuronal inactivation from neuronal loss, flumazenil-PET, an indicator of benzodiazepine receptor density, could add valuable information on the extent of brain damage. Activation studies focus on the evaluation of residual brain network, looking for processing in secondary projection fields. So far the predictive strength concerning possible recovery for the individual patient is limited, and PET and SPECT are not routine procedures in the assessment of patients in a vegetative state. PMID- 16350972 TI - Cerebral processing of auditory and noxious stimuli in severely brain injured patients: differences between VS and MCS. AB - We review cerebral processing of auditory and noxious stimuli in minimally conscious state (MCS) and vegetative state (VS) patients. In contrast with limited brain activation found in VS patients, MCS patients show activation similar to controls in response to auditory, emotional and noxious stimuli. Despite an apparent clinical similarity between MCS and VS patients, functional imaging data show striking differences in cortical segregation and integration between these two conditions. However, in the absence of a generally accepted neural correlate of consciousness as measured by functional neuroimaging, clinical assessment remains the gold standard for the evaluation and management of severely brain damaged patients. PMID- 16350973 TI - Residual auditory function in persistent vegetative state: a combined PET and fMRI study. AB - In recent years, a number of studies have demonstrated an important role for functional neuroimaging in the identification of residual cognitive function in persistent vegetative state. Such studies, when successful, may be particularly useful where there is concern about the accuracy of the diagnosis and the possibility that residual cognitive function has remained undetected. Unfortunately, functional neuroimaging in persistent vegetative state is extremely complex and subject to numerous methodological, clinical and theoretical difficulties. Here, we describe the strategy used to study residual auditory and speech processing in a single patient with a clinical diagnosis of persistent vegetative state. Identical positron emission tomography studies, conducted nine months apart, revealed preserved and consistent responses in predicted regions of auditory cortex in response to intelligible speech stimuli. Moreover, a preliminary functional magnetic resonance imaging examination at the time of the second session revealed partially intact responses to semantically ambiguous stimuli, which are known to tap higher aspects of speech comprehension. In spite of the multiple logistic and procedural problems involved, these results have major clinical and theoretical implications and provide a strong basis for the systematic study of possible residual cognitive function in patients diagnosed as being in a persistent vegetative state. PMID- 16350974 TI - Assessing level of consciousness and cognitive changes from vegetative state to full recovery. AB - Although investigations addressing cognitive recovery from the vegetative state have been reported, to date there have been no detailed studies of these patients combining both neuropsychology and functional imaging to monitor and record the recovery of consciousness. This paper describes the recovery of a specific vegetative state (VS) case. The patient (OG) remained in the vegetative state for approximately two months, increasing her level of awareness to a minimally conscious state, where she continued for approximately 70 days. In the course of the ensuing 18 months, she was able to reach an acceptable level of cognitive functioning, with partial levels of independence. Throughout this two year period, she received continuous cognitive evaluation, for which several different tools were applied including coma and low functioning scales, full cognitive batteries, and structural and functional magnetic resonance imaging (MRI). We present here preliminary data on fMRI using a word presentation paradigm before and after recovery; we also discuss the difficulty of how to determine level of consciousness using the tools currently available, and the subsequent improvement in different cognitive domains. We confirm that accurate diagnosis and proper cognitive assessment are critical for the rehabilitation of patients with disorders of consciousness. PMID- 16350975 TI - Neurophysiological correlates of persistent vegetative and minimally conscious states. AB - The evaluation of patients after severe brain injury is a complex process for the clinician, even with the information provided by a detailed neurological examination. The clinical examination often does not provide sufficient information to fully evaluate these patients due to several factors. Limited and inconsistent motor responses may obscure expression of greater cognitive capacities. More importantly, evaluation of the functional integrity of the cerebral cortical, thalamic and basal ganglia system is poorly indicated by the clinical examination in many patients. Neurophysiological studies provide a complementary set of objective data for evaluating brain-injured patients, as well as predicting and following the course of their recovery. This additional information can be of great importance since vegetative patients may be difficult to distinguish clinically from those in the minimally conscious state. This is important because the latter category of patients may have a significantly better prognosis for recovery in the initial phase of injury. Electrodiagnostic and imaging studies can help the practitioner to determine the degree of preserved and recovering neurological function. In this review we will assess the various neurophysiological studies currently at our disposal to evaluate and follow the clinical course of patients who have suffered severe brain injuries. PMID- 16350976 TI - Apallic syndrome is not apallic: is vegetative state vegetative? AB - Initial conceptualisation about the nature of vegetative state (VS) assumed at least temporary loss of the entire cortical functioning. Since a broad range of stimulus-related cortical activations was demonstrated in VS patients, this simplified idea is not tenable any longer, but no alternative concept emerges instead. Two recent hypotheses, empirically testable and well grounded, could fill this vacuum: (1) In VS, isolated cortical areas may work, but their integration into a distributed network is lacking. (2) In VS, complex stimulus processing is limited to primary sensory and motor areas; the co-ordination between them and the secondary and tertiary areas is lacking. To test these hypotheses, we estimated the frequency of occurrence of late event-related potential components P3 and N400, presumably indicating activity of complex distributed networks including high-level sensory and associative areas. Both components occurred in VS with above-chance frequencies, but less frequently than in two control groups. Besides these frequent normal brain activations, some VS patients exhibit highly significant but abnormal activations, whose functional meaning remains unclear. A methodological analysis leads to the conclusion that any neurophysiological assessment of VS patients is biased toward under-, rather than over-estimation, of their remaining information processing abilities. PMID- 16350977 TI - Neurophysiological patterns of vegetative and minimally conscious states. AB - This paper reviews the possible usefulness of electroencephalogram (EEG) and evoked potential (EP) recording in vegetative and poorly-responsive patients. There is a marked inter-individual EEG and EP variability, which reflects the state heterogeneity. Four clinical applications are described: (1) the identification of primary midbrain dysfunction--and, therefore, a possible reversibility--in post-traumatic states; (2) the identification of the permeability of sensory channels; (3) quantitative follow-up; and (4) individual assessment of cognitive functions and/or consciousness. Regarding this last issue, the loss of primary cortical EPs, although rarely observed, constitutes one major argument against consciousness. Conversely, cognitive EPs definitely proved the persistence of cognitive functions in several vegetative patients. Whether these cognitive functions are conscious or not remains a matter of debate. PMID- 16350978 TI - Evoked potentials for the prediction of vegetative state in the acute stage of coma. AB - For comatose patients in intensive care units, it is important to anticipate their functional outcome as soon and as reliably as possible. Among clinical variables the Glasgow Coma Score (GCS) and the patient's pupil reactivity are the strongest predictive variables. Evoked potentials help to assess objectively brain function. Over the past 20 years, numerous studies have assessed their prognostic utility in terms of awakening from coma. Fewer studies, however, have focused upon the utility of evoked potentials in predicting progression to the vegetative state. In this area evoked potentials appear to have a highly predictive value. In anoxic coma the abolition of somatosensory evoked potentials (SEPs) is related to a poor outcome, defined as death or survival in a vegetative state, with a 100% specificity. Following traumatic brain injury, the predictive value for unfavourable outcome is 98.5% when there are no focal injuries likely to abolish SEP cortical components. In contrast, the presence of event-related evoked potentials, and particularly mismatched negativity (MMN), is a strong predictor of awakening and precludes comatose patients from moving to a permanent vegetative state (PVS). PMID- 16350979 TI - Bispectral analysis of electroencephalogram signals during recovery from coma: preliminary findings. AB - The aim of this study was to investigate the accuracy of bispectral index (BIS), spectral edge frequency (SEF 95%), total power (TOTPOW) and frontal spontaneous electromyography (F-EMG) in monitoring consciousness in severely brain damaged patients. In 29 patients a total of 106 sedation-free and good quality EEG epochs were correlated with the level of consciousness as assessed by means of the Glasgow Liege Scale (GLS) and the Wessex Head Injury Matrix (WHIM). The strongest correlation with behavioural measures of consciousness was observed with BIS recordings. An empirically defined BIS cut-off value of 50 differentiated unconscious patients (coma or vegetative state) from conscious patients (minimally conscious state or emergence from minimally conscious state) with a sensitivity of 75% and specificity of 75%. These preliminary findings are encouraging in the search for electrophysiological correlates of consciousness in severe acute brain damage. PMID- 16350980 TI - Electrical treatment of reduced consciousness: experience with coma and Alzheimer's disease. AB - The right median nerve can be stimulated electrically to help arouse the central nervous system for persons with reduced levels of consciousness. The mechanisms of central action include increased cerebral blood flow and raised levels of dopamine. There is 11 years of experience in the USA of using nerve stimulation for acute coma after traumatic brain injury. There is a much longer period of experience by neurosurgeons in Japan with implanted electrodes on the cervical spinal cord for persons in the persistent vegetative state (PVS). But the use of right median nerve electrical stimulation (RMNS) for patients in the subacute and chronic phases of coma is relatively new. Surface electrical stimulation to treat anoxic brain injury as well as traumatic brain injury is evolving. Novel applications of electrical stimulation in Amsterdam have produced cognitive behavioural effects in persons with early and mid-stage Alzheimer's disease employing transcutaneous electrical nerve stimulation (TENS). Improvements in short-term memory and speech fluency have also been noted. Regardless of the aetiology of the coma or reduced level of awareness, electrical stimulation may serve as a catalyst to enhance central nervous system functions. It remains for the standard treatments and modalities to retrain the injured brain emerging from reduced levels of consciousness. PMID- 16350981 TI - Deep brain stimulation therapy for the vegetative state. AB - Twenty-one cases of a vegetative state (VS) caused by various kinds of brain damage were evaluated neurologically and electrophysiologically three months after brain injury. These cases were treated by deep brain stimulation (DBS) therapy, and followed up for over 10 years. The mesencephalic reticular formation was selected as a target in two cases, and the thalamic centre median parafascicular (CM-pf) complex was selected as a target in the other 19 cases. Eight of the 21 patients emerged from the VS, and became able to obey verbal commands. However, they remained in a bedridden state except for one case. DBS therapy may be useful for allowing patients to emerge from a VS, if the candidates are selected according to appropriate neurophysiological criteria. A special neurorehabilitation system may be necessary for emergence from the bedridden state in the treatment of VS patients. Further, DBS therapy is expected to provide a useful method in minimally conscious state (MCS) patients to achieve consistent discernible behavioural evidence of consciousness, and emergence from the bedridden state. PMID- 16350982 TI - Levodopa treatment for patients in persistent vegetative or minimally conscious states. AB - The persistent vegetative state (PVS) and the minimally conscious state (MCS) are conditions of altered consciousness after severe brain damage due to a variety of pathologies. However, the specific pathophysiological mechanisms and a therapeutic strategy for intervention have not as yet been established. We review previous reports of levodopa treatment for patients in PVS, MCS, or other mental disorders, and have focused on five representative cases: four of PVS and one of MCS after severe brain injury. In summary, our review suggests the effectiveness of levodopa treatment is probably dependent upon the following criteria: (1) Diagnosis of PVS or MCS as distinct from other related conditions, (2) Concomitant symptoms of parkinsonism, and (3) Concomitant neuroradiological findings of high intensity lesions in the dopaminergic pathway on T2 weighted MRI. The apparent success of levodopa in the five cases described may reflect a specific subgroup of PVS and MCS patients, where the administration of levodopa is effective. However, we should not regard PVS or MCS as a single entity, since levodopa is unlikely to be effective in all cases. Therapeutic strategies should aim to identify the key pathophysiological mechanism for each patient and target interventions accordingly. PMID- 16350983 TI - Part III: Behavioural assessment and rehabilitation techniques. PMID- 16350984 TI - Vegetative and minimally conscious states: serial assessment approaches in diagnosis and management. AB - Assessment of vegetative (VS) and minimally conscious state (MCS) patients presents clinicians with inherent difficulties (Royal College of Physicians, 2003) in terms of the reliable detection of potential signs of awareness given that all current assessment tools rely on observed behaviour. Recently developed measures such as SMART (Gill-Thwaites & Munday, 1999) and WHIM (Shiel et al., 2000), employing structured operational defined behavioural observations can facilitate the serial assessment of patient awareness, progress and appropriate goal setting particularly as one-off bedside assessments are more likely to be inaccurate. The use of sensitive tailored approaches involving experienced multidisciplinary teams is strongly advocated (Royal College of Physicians and British Society of Rehabilitation Medicine, 2003), notwithstanding clinicians should carefully consider potential confounding clinical factors, which may deleteriously influence patient arousal or ability to respond. Finally, areas for future development and recommendations regarding multidisciplinary assessment approaches with VS and MCS patients are outlined. PMID- 16350985 TI - The Disability Rating and Coma/Near-Coma scales in evaluating severe head injury. AB - The Disability Rating (DR) and Coma/Near-Coma (C/NC) scales for evaluating severe head injury are described. Scales are related to patient status, course and outcome and also underlying electroneurophysiological dysfunction. They lend themselves to high inter-rater reliability with brief training and can be completed in minutes, conserving staff time and energy. PMID- 16350986 TI - The JFK Coma Recovery Scale--Revised. AB - The JFK Coma Recovery Scale (CRS) was developed to help characterise and monitor patients functioning at Rancho Levels I-IV and has been used widely in both clinical and research settings within the US and Europe. The CRS was recently revised to address a number of concerns emanating from our own clinical experience with the scale, feedback from users and researchers as well as the results of Rasch analyses. Additionally, the CRS did not include all of the behavioural criteria necessary to diagnose the minimally conscious state (MCS), thereby limiting diagnostic utility. The revised JFK Coma Recovery Scale (CRS-R) includes addition of new items, merging of items found to be statistically similar, deletion or modification of items showing poor fit with the scale's underlying construct, renaming of items, more stringent scoring criteria, and quantification of elicited behaviours to improve accuracy of rating. Psychometric properties of the CRS-R appear to meet standards for measurement and evaluation tools for use in clinical and research settings, and diagnostic application suggests that the scale is capable of discriminating patients in the minimally conscious state from those in the vegetative state. PMID- 16350987 TI - Rehabilitation practice following profound brain damage. AB - The rehabilitation of the person with profound brain damage is a complex process requiring the skills of a true interdisciplinary team. The process is as much about the ability to: assess and diagnose; provide the optimal environment for recovery; prevent and treat secondary complications; support the family; and modify the environment as it is about a formal rehabilitation programme. Ideally these should be seamless but each contains many challenges, including the experience and skills of the observer, the ability to communicate with members of the family, and the ability to work within an interdisciplinary team. PMID- 16350988 TI - Rehabilitation outcome evaluation after very severe brain injury. AB - Few centres provide long-term therapy for survivors of very severe brain injury who continue in a minimally responsive state. We report on two outcome evaluation projects in association with one such centre in western Canada. In one project a functional scale to detect subtle changes after long-term therapy with the most severely compromised clients (Rancho levels II and III) is being tested. In the other project outcome indicators of change in quality of life after initiation of community-based rehabilitation have been generated by collecting over 400 critical incidents reported by family members, volunteers, staff and a few higher functioning clients. Our intention in this report is to highlight what can be done in terms of rehabilitation and outcome evaluation with clients who seem to be persisting in vegetative or minimally responsive states. PMID- 16350989 TI - Rehabilitation interventions for vegetative and minimally conscious patients. AB - Brain injury rehabilitation is a complex and challenging task for all members of the multidisciplinary team. Medical advances have allowed more severely impaired patients to survive and consequently the number of patients in the vegetative and minimally conscious states have proportionately increased. Thus, the need for evidence-based practice and further research demonstrating the effects of specific rehabilitation interventions is required. This article reviews the current research and consensus on rehabilitation for patients in the vegetative and minimally conscious states. PMID- 16350990 TI - Can behaviours observed in the early stages of recovery after traumatic brain injury predict poor outcome? AB - Diagnosis of the vegetative state (VS) or minimally conscious state (MCS) is dependent on the presence or absence of behavioural variables. Diagnosis of the VS in the case of those who have had a severe head injury does not occur for several months after injury. If such outcomes could be predicted it would facilitate appropriate and timely referrals to specialist rehabilitation units. This paper describes a follow up study of patients observed prospectively after severe head injury and followed up four years later. Thirty patients had made a moderate or good recovery and eight met criteria for MCS. Analysis of early observations together with outcome data suggest that the duration of time taken to achieve some early behaviours may be predictive of poorer outcome. PMID- 16350991 TI - Vegetative and minimally conscious states: how can occupational therapists help? AB - There is little documentation about the role of occupational therapy specifically for the vegetative and minimally conscious patient. This paper sets out to clarify the role using one model of occupational therapy, namely that of Reed and Sanderson (1992), and proposes occupational therapy assessment and treatment for this patient group. It explores how patients are affected in the performance components of motor, sensory, cognitive, intrapersonal, and interpersonal skills, which are explored through assessment. The paper then explains how these skills impact upon occupational performance (functional ability) and suggests methods occupational therapists will use in treatment planning. Throughout the manuscript, a person-centred approach, which is vital to occupational therapy, is demonstrated. PMID- 16350992 TI - Considerations for the use of assistive technology in patients with impaired states of consciousness. AB - While there is limited literature addressing the application of assistive technology in patients in persistent vegetative state (PVS) and minimally conscious state (MCS), it is believed that it can assist with the assessment, diagnosis and treatment as well as management of these patients. The use of technology to assist in PVS and MCS is mostly limited to the application of simple binary switch devices to determine whether a motor response is consistent or otherwise. However, the application of such technology is often undermined due to a lack of established protocols for use by the multidisciplinary team (MDT), as well as a lack of available technical resources. Therefore the ongoing development of assessment instruments as well as effective outcome measures used by an MDT is imperative. This article aims to discuss some key aspects to consider in the use of assistive technology when assessing and treating people in impaired conscious states. Possible considerations and suggestions will be discussed through this paper and a case study will be used to demonstrate some of these interventions. PMID- 16350993 TI - Music therapy with patients in low awareness states: approaches to assessment and treatment in multidisciplinary care. AB - This paper outlines the rationale for and role of music therapy as a clinical intervention and diagnostic tool in multidisciplinary (MDT) rehabilitation programmes for patients in low awareness states. A review of the literature indicates that music is a useful clinical tool in stimulating a range of behavioural, physiological and expressive responses in patients in low awareness states. Referral criteria for music therapy with this patient group are provided, along with suggested methods for collaborative multidisciplinary work. A case vignette is presented of a client whose diagnosis of vegetative state (VS) was contradicted by her purposeful responses within music therapy assessment, contributing towards a changed diagnosis to minimally conscious state (MCS). The case illustrates the particular role of music therapy in assisting with diagnosis in complex cases. Music therapy provides a clinical forum in which recovery of function can be assessed in an informal way, using a medium which does not rely on language, is non-evasive and elicits emotional responses. PMID- 16350994 TI - Nutrition and hydration for the vegetative state and minimally conscious state patient. AB - This paper presents nutritional issues particular to patients in the vegetative state (VS) or minimally conscious state (MCS). It assumes that such patients would be tube fed and it examines suitable ways of assessing and monitoring their nutrition. It covers problems frequently encountered such as undernourishment, high fluid requirements, bowel management, and vomiting. It also looks at the practicalities of long-term tube feeding. Once medical stability has been achieved, feeding these patients is almost always successful with the patient's body weight restored to being within normal limits. PMID- 16350995 TI - Psychological needs of patients in low awareness states, their families, and health professionals. AB - Patients who have emerged from low awareness states may present with psychological needs that can be addressed via adapted formal clinical psychological interventions, or by behavioural techniques. Families of these patients may experience similar psychological reactions to relatives of any patients with severe brain injury, but there are also additional factors that are unique to patients in low awareness states. These sources of psychological distress for relatives are discussed. The needs of clinicians working with these clients are also discussed. It is important that services attending to the needs of clients in low awareness states also have adequate support for both relatives and clinicians. PMID- 16350996 TI - [Displacement of the instantaneous center of rotation of the humeral head during abduction: implication for scapulohumeral muscular function]. AB - PURPOSE OF THE STUDY: To date, studies of glenohumeral stress forces have been based on the notion of a unique center of rotation situated at the geometric center of the humeral head. Early work, e.g. Fisher et al in 1977, suggested that the instantaneous center of rotation can migrate during abduction producing muscle vectors with variable moments and consequently variable stress forces. We conducted a kinematic analysis of the shoulder forces during continuous movement. MATERIAL AND METHODS: An imaging device commonly used for angiography enabled acquisition of one image per second during a continuous movement of abduction. Images were obtained for twenty healthy shoulders in ten subjects. RESULTS: The instantaneous center of rotation was not situated at the geometric center of the humeral head. Although interindividual variability was significant, the preferential positions of the instantaneous center of rotation implied a constant reproducible succession of gliding, rolling, and translation in variable proportions during the abduction movement. When the same movement was performed with a 3500g load held in the hand, the ICR migrated towards the upper part of the humeral head implying a predominance of the rolling motion over the gliding and translation motions. Between 40 degrees and 60 degrees glenohumeral abduction, the instantaneous center of rotation exhibited a metaphyseal displacement producing a longer lever arm for the supaspinatus and an abduction function for the infraspinatus and the subscapularis, unrecognized to date. After 60 degrees, medialization of the instantaneous center of rotation lengthened the lever arm of the deltoid. DISCUSSION: A valid analysis of the glenohumeral stress forces must take into consideration the localization of the instantaneous center of rotation. In the clinical setting, our findings demonstrate that the infraspinatus and the subscapularis play an important role which must be considered both in surgery and in rehabilitation. PMID- 16350997 TI - [Recent fractures of the radial head associated with elbow instability treated with floating Judet prosthesis]. AB - PURPOSE OF THE STUDY: Fracture of the radial head associated with elbow instability is infrequent. We report a retrospective series of floating Judet prostheses implanted for comminutive fractures of the radial head associated with elbow laxity caused either by dislocation or rupture of the medial collateral ligaments. MATERIAL AND METHODS: The series included ten patients who underwent surgery from October 1996 to September 2002 at the Amiens University Hospital. The indication for radial head prosthesis was established in the emergency setting for fracture unamendable by osteosyntheis and elbow laxity. Mean age was 48.2 years (25-69). All patients were seen at mean follow-up of 31.7 months (18 48). According to the Mason classification as modified by Johnson, all patients had type 4 fracture. A Judet radial head prosthesis with a floating metallic cup was implanted in all patients. An investigator other than the operators evaluated outcome using the Mayo Clinic criteria. RESULTS: Joint motion as measured by goniometry was: mean flexion 121degrees (90-140 degrees), mean extension deficit 20 degrees (5-60 degrees), mean pronation 45 degrees (0-85 degrees), mean supination 42.5 degrees (0-90 degrees). The Mayo clinic score was excellent in 3, good in 2, fair in 3, and poor in 2 (prosthesis dislocation in one and hung prosthesis requiring removal in one). Four patients developed periarticular ossifications compromising the final result. DISCUSSION: The floating Judet prosthesis allows optimal adaptation of the implant to anatomy and function. For us, these implants are indispensable when the radial head fracture is associated with elbow instability. The indication for prosthesis may be questionable if the non-fixed fracture is free of associated ligament injury, as suggested by the good long-term reported after simple resection. Rigorous operative technique is crucial, with careful restitution of the radial height. The quality of the result is related to the degree of capsule and ligament injury even after optimal implant positioning. Preventive treatment against periarticular ossification should be systematic. PMID- 16350998 TI - [Excision of heterotopic new bone around the elbow in patients with head injury: 51 cases]. AB - PURPOSE OF THE STUDY: Neurogenic paraosteoarthropathy of the elbow is a frequent complication in head injury patients. The functional impairment may be severe. The purpose of this work was to evaluate the efficacy of our medico-surgical approach and to determine indications for surgery. MATERIAL AND METHODS: We analyzed retrospectively a continuous series of 44 patients (51 elbows) with neurogenic heterotopic new bone of the elbow joint managed from 1993 to 2000. The heterotopic bone had developed on a central deficit limb in 70% of the patients. Flexion-extension was limited in all patients, and pronation-supination in nine. The ulnar nerve was compressed by the heterotopic bone in half of the patients. A 3D reconstruction CT-scan with contrast injection was obtained in all patients. We consider did not the classical criteria for bone maturation to assess operability. Using an adapted approach, we removed the heterotopic new bone to enable complete elbow motion. Associated procedures were performed in fifteen patients due to persistent intraoperative joint stiffness related to muscle tendon retraction. We did not prescribe diphosphonates or radiotherapy postoperatively. RESULTS: At 45 months follow-up, one patient had been lost. Relative gain was considered very good in 34 elbows, good in 13 and fair in 3. There were no cases of lost motion. There were no recurrences causing joint limitation, but two of nine cases of radioulnar synostosis recurred. DISCUSSION: Neurogenic paraosteopathy of the elbow impairs joint function and may lead to ulnar nerve compression. The goal of surgery is to improve function. The analysis of our results show that early surgery yields good results without complementary treatment, irrespective of the patient's overall functional deficit or cognitive status and irrespective of the localization of the hetertopic bone. The essential prognostic factor for the quality results is the neurological status of the limb, particularly the degree of spasticity and muscle-tendon retraction. PMID- 16350999 TI - [Prospective study assessing quality-of-life after hemiarthroplasty for hip fracture]. AB - PURPOSE OF THE STUDY: We conducted a prospective multicentric study of 203 consecutive patients who underwent hemi-arthroplasty for femoral neck fracture between May 1999 and August 2001. The purpose of the study was to assess quality of-life among these patients one year after their surgery. MATERIAL AND METHODS: Mean patient age was 79.6 +/- 8.6 years; 71.2% were women. Most fractures were Garden type III and IV (87.4%) and 90.9% were recent (< 21 d). The hemiarthroplasty was performed with the Intermedia implant via a posterolateral approach in 73.9% of patients with acetabular reaming for 13.7%; a skirted head was used in only 20.8%. The Robinson score (19/26 < or = 6.89) for joint motion, lifestyle, degree of osteoporosis (Singh score), and history (ASA score), as well as a psychomotor indicator (Hodkinson score) were noted preoperatively. Quality of-life was measured at one year in the same patients using the Postel Merle d'Aubigne (PMA) score and a quality-of-life score. RESULTS: Among the 203 patients, 38 (18.7%) were lost to follow-up and 37 died (18.2%). Eleven posterior dislocations (5.4%) were noted and four were reduced orthopedically. The PMA score was 84.7% satisfactory results (excellent, very good, good). Preoperatively 58.8% of the patients were independent and 7% were in nursing homes; one year postoperatively, these proportions were 54% and 4.8%. There was a significant difference in the PMA score between two populations defined by the preoperative Robinson score less than or greater than 20. Outcome was better for patients with a Robinson score > 20 pre-operatively. For these two populations, change in lifestyle was highly significant one year postoperatively. However, for the overall population, changes in lifestyle and joint motion were not significantly different between the pre- and postoperative evaluations. Paradoxically, 29.1% of the patients who were dependent preoperatively were not one year after surgery, which indicates that certain patients also had a better quality-of-life after their hemiarthroplasty. CONCLUSION: Mortality after fracture of the femoral neck remains high (18.2% at one year). There were no infections requiring revision in this series, but there were eleven cases of dislocation (5.4%). The difference in joint motion and quality-of-life between the pre- and postoperative assessment was not significant for the whole population, but one third of the patients who were independent preoperatively were not at last follow-up. PMID- 16351000 TI - [Reliability of porous coating metal-backed cups: advantages and adverse effect of a posterior elevated rim polyethylene liner]. AB - PURPOSE OF THE STUDY: We analyzed primary implantation of the Duraloc cup associated with a self-locking Muller stem at 9.6 +/- 1.6 years (8.6-11.4). Because of its particular 3/5 semi-spherical shape, the Duraloc cup has been associated with a high rate of dislocation. The objective of this study was to evaluate the efficacy of an anti-dislocation posterior elevated rim polyethylene liner on long-term dislocation rate and wear. MATERIAL AND METHODS: The series included 89 arthroplasties implanted between 1991 and 1993 in 82 patients, mean age 56.8 +/- 12.6 years (17.2-87). The Postel-Merle-d'Aubligne (PMA) score and subjective evaluation with a visual analog scale were used for the clinical assessment. Changes in the bone-cup interface, cup migration, and polyethylene wear were assessed radiographically. RESULTS: At last follow-up, the PMA function score was 16.2 +/- 1.9 points (10-18) (81% good, very good and excellent results). The postoperative x-rays showed a low incidence of lucent lines and osteolysis, respectively 8% and 4%. Mean annual linear polyethylene wear was 0.11 +/- 0.066 mm (0.03-0.57) and only 4% of the cups showed wear greater than 0.2 mm/year. Wear was correlated with the presence of acetabular osteolysis and high activity level. Two cups migrated (3-4 cm medialization). Three cups were revised, one for deep infection, one for recurrent dislocation, and one for dislocation associated with major wear. The rate of dislocation was 2.2% (two cases). Overall survival was 97.3% at 9.6 years (95%CI 0.93-1.0). DISCUSSION: Compared with other series in the literature using this implant, the presence of a posterior elevated rim polyethylene liner reduced the rate of instability and did not increase wear. The Duraloc cup is recognized as a reliable implant exhibiting excellent osteointegration and a low rate of migration. The presence of a posterior rim is associated with a lower rate of dislocation and does not increase wear. It can thus be proposed for primary implantation. The posterosuperior position for the liner is recommended. PMID- 16351001 TI - [Early femoral osteolysis following Charnley-Kerboull total hip arthroplasty combining a 22-mm zirconia head on polyethylene]. AB - PURPOSE OF THE STUDY: New bearings have been developed to reduce polyethylene wear. Zirconia ceramic is one proposal with attractive tribologic properties. The purpose of this prospective study was to evaluate the clinical and radiological results at least two years after implantation in a continuous series of total hip arthroplasties combining a zirconia head with polyethylene socket. MATERIAL AND METHODS: The series included 56total hip arthroplasties performed in 51 patients (30 females and 21 males), mean age 52.2 +/- 12 years (25-76 years). Prostheses were implanted for primary degenerative disease (43%) and dysplasia (27%). The femoral component was a stainless steel stem with a Morse cone measuring 11degrees 25' for 27 hips and 5 degrees 40' for 28 hips. A 22-mm stabilized yttrium tetragonal polycrystalline zirconia head (Y-TZP) was used. The transtrochanteric approach was used for all arthroplaties with implantation of a cemented Charnley-Kerboull prosthesis. Clinical outcome was assessed with the Postel-Merle-d'Aubligne (PMA) score. Acetabular cup wear and periprosthetic osteolysis were measured on successive AP x-rays of the pelvis. RESULTS: Mean follow-up was 32 months (24-48). None of the patients were lost to follow-up. Mean function score at last follow-up was 17.8 +/- 0.2 (16-18) versus 12.2 +/- 2.6 preoperatively (Wilcoxon, p < 0.0001). There were no cases of implant migration (femoral or acetabular). An endosteal defect by femoral osteolysis in the calcar femoral was observed in 19 of 55 cases. It appeared early between first and second post-operative year and did not progress later. Its surface remained less than 1 cm2 in all cases. We were unable to identify any factor predictive of these osteolytic lesions. DISCUSSION: The short-term clinical results with this type of arthroplasty was comparable with earlier results described with classical Charnely-Kerboull implants using a metal-polyethylene bearing. The Merkel osteolysis observed in this series occurred early for one third of the hips with no measurable polyethylene wear. We suggest surgeons should be cautious about using the zirconia head. We are continuing our surveillance of these patients. PMID- 16351002 TI - [Single-plane external fixation of fresh fractures of the femur: critical analysis of 53 cases]. AB - PURPOSE OF THE STUDY: External fixation has not been widely used for femoral fractures and few series are reported in the literature. External fixation is generally reserved for severe open fractures, for vessel injury or multiple trauma with life threatening. We present a retrospective analysis of a serie treated in a single center in order to detail the indications of this fixation technique. MATERIAL AND METHODS: From 1984 to Jun 2002, 49 patients with femoral fractures were treated by external fixation. The series included 36 men and 13 women, mean age 31 years. All were victims of high-energy trauma: traffic accident (n = 40), fall from high level (n = 4), firearm wound (n = 5). Multiple fractures were present in all patients except seven and 24 patients had multiple injuries. Forty fractures were open fractures: two type 1, ten type 2, four type 3a, 23 type 3b and five type 3c in the Gustilo classification. Twenty-seven were shaft fractures and 26 involved the distal metaphyseoepiphyseal portion of the femur. Loss of cortical stock was noted in five cases and total loss of a segment in four. Surgery was deferred in 19 patients, mean six days. A single-plane external fixation was used (Orthofix) with a femorofemoral frontolatateral assembly. Transepiphyseal screw fixation was also used to stabilize the distal fracture in eleven cases. RESULTS: One patient with a bifocal fracture of the femur died from head trauma. Three patients required above knee amputation after failure of a vessel bypass or due to septic necrosis of the reconstruction flap. Five patients required a second reduction within days of external fixation. On the AP view, femoral alignment was successfully reestablished at +/- 5 degrees in 45 cases, ranged from 5 degrees to 10 degrees in seven and was greater than 10 degrees in one. On the lateral view, alignment was between 5 degrees and 10 degrees in 42 cases and greater than 10 degrees in one. Femur length was equal to the healthy side in 23 cases, and was shortened 1-2 cm in 26. Four metaphyseal fractures resulted in a 3 cm shortening. Bone healing time was available for 42 patients (1 death, 3 amputations, 3 lost to follow-up). Elective conversion to internal fixation was performed in ten patients (five lateral cortical plates and five centromedullary nailings). These patients all achieved first-intention bone healing with a mean time of 7.4 months. Exclusive external fixation was planned for 34 fractures. First-intention healing was achieved in 25 (17 shaft and 8 distal) without bone graft with an average time of 7.3 months. Ten patients had one or more osteitis foci on pin tracts. Two patients in this group developed recurrent fracture after removal of the external fixator. Nine fractures did not heal and required revision with centromedullary nailing (n = 5) or plate fixation with autograft (n = 4). Nailings for nonunion were successful but plate fixation was compromised by infection in one patient and recurrent fracture after plate removal in another. Fourteen patients underwent joint mobilization under general anesthesia and 14 had open arthrolysis. Mean follow-up was 2.8 years. Mean active flexion was 90 degrees (30-130 degrees). Ten patients exhibited flexion between 30 degrees and 60 degrees and 19 between 70 degrees and 100 degrees. Knee flexion was greater than 110 degrees in 15 patients. Residual 10 degrees flexion was noted in six knees. Mean leg length discrepancy was 0.4 +/- 0.6 after distal fracture and 0.8 +/- 1.3 after diaphyseal fracture. DISCUSSION: The indications and results of external fixation in this series are in line with reports in the literature. For diaphyseal fractures, healing is long and difficult, partly because of the insufficient mechanical properties of external fixation. The rate of infection and stiff knee is high, particularly for distal fractures of the femur. CONCLUSION: External fixation remains the only solution to stabilize certain open diaphyseal fractures or for patients with life-threatening multiple injuries. This techniques allows control of the other traumatic lesions while waiting for internal fixation. For fractures of the distal femur, external fixation can only be advocated for metaphyseodiaphyseal fractures with an intact or reconstructed epiphyseal portion. PMID- 16351003 TI - [Discoid menisci in children: clinical patterns and treatment in eighteen knees]. AB - PURPOSE OF THE STUDY: Surgery is indicated when discoid menisci become symptomatic. The purpose of the surgical procedure is to resect any damaged tissue and restore the physiological shape of the meniscus. As a rule, as much meniscal tissue as possible must be spared because of the long-term deleterious effects on the cartilage of total meniscectomy. We analyzed a consecutive series of 18 cases of discoid menisci in children to search for secondary lesions and factors favoring their development in order to determine the optimal surgical procedure. MATERIAL AND METHODS: This retrospective series included 17 children (18 menisci) aged 7.5 years on average at diagnosis between 1985 and 2003. We noted the clinical manifestations, the imaging findings, time to treatment, and operative observations. The Watanabe classification was used to describe the discoid menisci. We also noted meniscal and cartilage damage and their consequences, as well as the consequences of late surgery on the type of procedure used. RESULTS: The main complaint was pain. Physical examination usually revealed a positive pivot test. Signs of osteochondral lesions of the lateral condyle were also observed in three children, and the MRI revealed degenerative menisci in four. Mean time from diagnosis to surgical treatment was 20 +/- 17 months. The Watanabe classification was type I (n=9), type II (n=5), type III (n=4). Arthroscopy revealed nine meniscal lesions and three cartilage lesions, one associated with osteochondritis. Meniscectomy was performed in eleven cases, meniscoplasty in seven. Meniscectomy was significantly more frequent (p<0.05) when there was a meniscal lesion (9/11 of the meniscectomies) and when the time from diagnosis to treatment was long (28 months versus 8 months for meniscoplasty, p<0.01). Time to surgery was associated significantly (p<0.05) with the proportion of meniscal, chondral, or osteochondral lesions. Inversely, the type of meniscus did not affect age at diagnosis, initial manifestations, or presence of a positive pivot test. DISCUSSION: While therapeutic abstention is warranted for asymptomatic menisci, surgical treatment should be undertaken if symptoms develop, irrespective of the type. If possible, surgery should be performed less than six months after diagnosis. The risk of secondary meniscal or cartilage injury increases with longer delay before surgery. Similarly, the chances of performing meniscoplasty are reduced with longer time from diagnosis to surgery. It must be recalled that the objective of conservative mensical surgery is to prevent secondary cartilage lesions after extensive meniscectomy and consequently the risk of osteoarthritis. PMID- 16351004 TI - [Percutaneous compression plate for pertrochanteric fixation of hip fractures: surgical technique and first results]. AB - We have used a minimally invasive technique for fixation of trochanteric fractures since 2003. We use the percuntaneous compression plate described by Gotfried. We describe here the osteosynthsis technique and our special approach which limits operative time. This technique avoids wide opening of the aponeurosis of the tensor fascia lata, major detachment, and section of the vastus lateralis. The plate is inserted via a 2-cm incision over the greater trochanter. Head and shaft screws are inserted via a second incision measuring 3 to 4 cm. Perfect reduction before plate insertion is the key to success. The results of our first twenty cases have demonstrated an uneventful postoperative period and a short operative time of 25 minutes on average. Peri-operative bleeding has been very limited. Radiographic healing was obtained in three months. Weight bearing was possible immediately after fixation in five cases and was delayed in fifteen. There were two early displacements which were analyzed. PMID- 16351005 TI - [Arthroscopic resection of pigmented villonodular synovitis pseudotumor of the shoulder: a case report with three year follow-up]. AB - Pigmented villonodular synovitis can be localized or diffuse. Lesions predominate in the knee but all of the joints can be involved. Thirty cases in the glenohumeral joint have been reported in the literature. The different reports to date have not identified any specific clinical signs. Our female patient presented non-specific shoulder pain which persisted for three years. The plain x rays were normal. MRI and arthroscan revealed an intra-articular pseudotumor. Arthroscopy was performed for tumor biopsy which was followed by total resection. The diagnosis of villonodular synovitis pseudotumor suspected at arthroscopy was confirmed at the pathology examination. The functional outcome was excellent and no recurrence has been observed at three years follow-up. Arthroscopy is less aggressive than open surgery for arthrotomy. Arthroscopy must be performed for diagnostic purposes since imaging findings are not specific. Arthroscopic synovectomy is the treatment of choice for pigmented villonodular synovitis in both the diffuse and pseudotumor forms. PMID- 16351006 TI - [Rupture of the flexor tendons on an anterior plate for distal radial fracture: four cases and a review of the literature]. AB - We report four cases of flexor tendon ruptures which occurred after distal radial fracture and reviewed the 25 other cases reported in the literature since 1932. Analysis of these 29 cases disclosed the causes of these ruptures. A deformed callus after distal radial fracture or presence of an anterior osteosynthesis plate can under certain conditions lead to secondary flexor tendon tears. It was also found that tears of the flexor pollicis longus rupture predominate, followed by injury to the flexor digitorum profundis and superficialis of the index finger. Other tendons have only been involved in only a few cases. In order to avoid this complication, we propose systematic removal of anterior plates or secondary replacement if the reduction is not totally anatomic. Surgeons should recall the importance of anatomic reduction of distal radial fractures. PMID- 16351007 TI - [Long-term follow-up after leg replantation]. AB - Leg replantation is generally considered to yield good results, particularly since the widespread use of progressive lengthening procedures. This optimism must be weighted against the very long and difficult operative program, the social and occupational implications, and the functional outcome of the replanted limb. We report a case of below knee leg replantation in a young patient whose operative program lasted 29 months. Occupational activities were interrupted for three years. The clinical result was satisfactory but with 14 degrees residual valgus suggesting a potential risk of osteoarthritis and difficult therapeutic decisions. The alternatives to replantation are controversial. Leg transplantation is not a valid option in our opinion. Amputation with orthesis should be considered more often because complications are rare and recovery much shorter. Indications for leg replantation should be considered with prudence. PMID- 16351008 TI - [In reference to the article: "Nerve compression caused by mucoid pseudocysts: arguments favoring an articular etiology (report of 23 cases)"]. PMID- 16351009 TI - [Hearing and the skin--a functional connection]. PMID- 16351010 TI - [Adjunctive treatments for acne therapy]. AB - The mainstays of modern acne therapy include comedolytic, antimicrobial, and anti inflammatory substances, as well as antiandrogens. Additionally, traditional or newly developed therapeutic approaches may be considered, including pharmacologic (dapsone, zinc) and physical measures (phototherapy, photodynamic therapy, comedone extraction, abrasives). This article reviews such adjunctive therapies with regard to efficacy and their roles in acne therapy. PMID- 16351011 TI - [MCTD--mixed connective tissue disease]. AB - Mixed connective tissue disease is a disease entity characterized by overlapping symptoms of lupus erythematosus (LE), systemic sclerosis (SSc), polymyositis/dermatomyositis (PM/DM) and rheumatoid arthritis (RA). Diagnostic criteria include high titers of antibodies against U1RNP as well as the presence of at least 3 of 5 of the following clinical features: edema of hands, synovitis, myositis, Raynaud phenomenon and acroscierosis. In terms of the pathogenesis, genetic as well as infectious (viral) factors appear to play a role. The acceptance of MCTD as a distinct disease entity is controversial. Terms such as "undifferentiated connective tissue disease" or "overlapping syndromes" are not helpful. One-quarter of MCTD patients transform into LE, while one-third progress to SSc. Therapeutic recommendations are glucocorticoids in combination with immunosuppressive agents and endothelin receptor antagonists. Double blind studies are not available. The prognosis is relatively good. Causes of death include pulmonary hypertension, infections and both pulmonary and cardiac failure. PMID- 16351012 TI - [Connexin 26 mutation and keratitis-ichthyosis-deafness (KID) syndrome]. AB - BACKGROUND: Keratitis-ichthyosis-deafness syndrome (KID syndrome) is an extremely rare disorder. Inheritance is autosomal dominant but many cases occur sporadically following a spontaneous mutation. The cause of KID syndrome are missense mutations of the gene GJB2, encoding connexin 26. PATIENTS AND METHODS: We clinically studied two cases of KID syndrome and extracted genomic DNA from peripheral blood. RESULTS: The patients showed different heterozygous mutations of the connexin 26 gene and had quite different clinical courses. CONCLUSIONS: Both patients showed heterozygous mutations of the connexin 26 gene; a different Cx26 dominant mutation can cause a very different clinical course. PMID- 16351013 TI - Cosmetic results of histographically controlled excision of non-melanoma skin cancer in the head and neck region. AB - BACKGROUND: Beside the primary goal of complete eradication, the cosmetic result is an important aspect of the treatment of non-melanoma skin tumors especially in the head and neck region. PATIENTS AND METHODS: From 1990 to 2000, we treated a total of 5,227 large basal cell carcinomas (BBC) and 1,189 squamous cell carcinomas (SCC) in the head and neck region by surgical excision in 4,239 inpatients at the Department of Dermatology, University of Tubingen. The procedure used in all patients was a conservative excision controlled by complete three dimensional histology of all margins (3D-histology) and specifically targeted follow-up surgery where required (histographic surgery). As part of the prospective tumor follow-up, we asked the treating outdoor physician one and four years later to evaluate the results of our surgical procedures. RESULTS: Of the 5,565 follow-up questionnaires sent back, 4,868 contained answers regarding the cosmetic result. The data from both answers were pooled. In 1,972 (40,5 %) patients the cosmetic result was evaluated as "excellent", in 1,992 (40,9%) as "good", in 662 (13,6%) as "satisfactory", in 191 (3,9%) as "mediocre" and in 51 (< 1,0%) as "poor". In 697 of the responses, the physician did not comment the cosmetic results or the patient was lost for follow up. CONCLUSION: With respect to both long term safety and cosmetic outcome, tumor surgery with 3D-histology of excisional margins has set very high quality standards in the treatment of non melanoma skin cancer of the head and neck area. PMID- 16351014 TI - [Ulceration of the scalp: a unique manifestation of pyoderma gangrenosum]. AB - Pyoderma gangrenosum is a chronic, noninfectious, ulcerating cutaneous disease of unknown etiology usually presenting with multiple lesions on the legs and trunk. Solitary lesions in other locations are rare and therefore hard to diagnose. A 78 year-old and a 81-year-old woman with extensive scalp necrosis were both diagnosed as pyoderma gangrenosum after exclusion of other differential diagnostic possibilities. The separate diagnosis of malignant pyoderma for this disease state no longer seems justified. Scalp necrosis is infrequently seen in dermatological patients; most previous reports describe it in giant cell arteritis. Other differential diagnoses include viral, bacterial and mycotic infections as well as erosive pustular dermatosis of the scalp. In conclusion, we again draw attention to unilesional pyoderma gangrenosum of the scalp as a rare disease entity. Immunosuppressive drugs are an effective treatment modality. PMID- 16351015 TI - [Acral scleroderma presenting simultaneously with small-cell bronchial carcinoma: a paraneoplastic disease?]. AB - The possible causal connection between systemic scleroderma and malignant neoplasms is a controversial issue. In the case of dermatomyositis, a malignant tumor is viewed as a possible trigger of the collagen disease; in contrast, scleroderma is suspected of causing tumors because of the long-term tissue fibrosis. A 68-year-old woman presented with acral scleroderma but already had metastatic bronchial carcinoma without evidence for previous pulmonary fibrosis. The rapid spread of the tumor and the sudden appearance of scleroderma suggest that in this case scleroderma could represent a paraneoplastic syndrome. Attention is directed to several growth factors and cytokines. On the one hand they play a part in the neoplastic transformation of the host cells and on the other hand they can change fibroblasts into a profibrotic phenotype and induce fibrosis. Impressive in this case is the fact that acral scleroderma definitely developed after the malignant tumor but before treatment, so that both fibrosis of the lung and side effects of chemotherapy and radiation can be excluded as triggers. PMID- 16351016 TI - [Occupational skin protection]. PMID- 16351017 TI - [90-year-old male with a hyperkeratotic lichenoid skin eruption]. PMID- 16351018 TI - [Patient information: Hypertrophic (severe) anal papillae]. PMID- 16351019 TI - [Continuing education lecture program of the VOD and DDG]. PMID- 16351020 TI - Methicillin-resistant Staphylococcus aureus pneumonia in children: a call for increased vigilance. PMID- 16351021 TI - Community-acquired MRSA: a notable adversary. PMID- 16351022 TI - Psychosomatic reasons for chronic pains. PMID- 16351023 TI - Treatment of late stage non-small cell lung cancer: we have more work to do. PMID- 16351024 TI - Malpractice claims: finding the silver lining. PMID- 16351025 TI - Salmonella infections in the setting of AIDS: a serpentine course. PMID- 16351026 TI - Helicobacter pylori treatment. PMID- 16351027 TI - Widespread emergence of methicillin resistance in community-acquired Staphylococcus aureus infections in Denver. AB - BACKGROUND: Increasing rates of methicillin resistance among out-patient Staphylococcus aureus infections led us to assess the epidemiology and outcome of a local outbreak. METHODS: A retrospective cohort study of outpatient skin and soft tissue infections due to S aureus in 2003. RESULTS: From 2002 to mid-2004, the percentage of outpatient S aureus isolates resistant to methicillin increased from 6 to 45%. In multivariate analysis, only male sex and age greater than 18 years were associated with methicillin resistance. Methicillin resistance was common (>15%) among isolates from patients in nearly all subgroups evaluated. Pulsed field gel electrophoresis showed isolates related to USA 300, but methicillin-resistant strains had unusually high rates of quinolone resistance. CONCLUSIONS: A single strain of methicillin-resistant S aureus is responsible for the increase in skin infections in outpatients without traditional risk factors for infection with an antibiotic-resistant strain. In areas with high rates of methicillin-resistant S aureus outpatient infections, we recommend non-beta lactam antibiotics for initial treatment of skin and soft tissue infections. PMID- 16351028 TI - Analysis of diagnostic error in paid malpractice claims with substandard care in a large healthcare system. AB - OBJECTIVE: Although claims databases are not representative of all care delivery, their predisposition toward serious unintended injury can complement resource intensive chart reviews and guide patient safety initiatives. MATERIALS AND METHODS: Non-Veterans Health Administration (VA) practitioners reviewed 1,949 VA malpractice claims paid during fiscal years 1998 through 2003. The portion associated with substandard care, the severity of harm, and types of negligence were identified. RESULTS: Negligent adverse events occurred in 37% (n = 723) of paid VA malpractice claims. These had high proportions of serious injury (55%) and morbidity (37%). Diagnostic negligent adverse events were most frequent (45%) and with 41% associated morbidity. The annual incidence of diagnosis-related paid VA malpractice claims was 1.95 per 100,000 patients and predicts that 122 of every 100,000 patients may have diagnostic negligent adverse events. Comparisons against non-VA data suggest this to be a healthcare industry problem. CONCLUSIONS: Diagnosis-related negligent adverse events are a serious problem in the healthcare industry. PMID- 16351029 TI - Late stage (III and IV) non-small cell cancer of the lung: results of surgical resection at Inova Fairfax Hospital. AB - One hundred forty-two patients underwent surgery and related treatment for advanced stage (III, IV) non-small cell cancer of the lung. One hundred seventeen patients underwent up-front surgery, with a hospital mortality rate of 1.7% (2/117). Kaplan-Meier 5-year survival in this group was 31% (+/- 5). Twenty-five patients underwent neoadjuvant therapy followed by surgical resection, with respective rates of hospital mortality, complete pathologic response, and major pathologic response of 0%, 16%, and 64%. Kaplan-Meier 5-year survival in this latter group was 34% (+/- 11). Of the 16 patients undergoing neoadjuvant therapy who had complete pathologic response or significant downstaging from stage III disease, Kaplan-Meier 5-year survival was 61% (+/- 15). Three clinical observations of interest emerged regarding survival. First, in those patients with postresection FEV1 < 1.0 L, hospital mortality rate was 20%, and there were no 5-year survivors (P < 0.0001). Second, where neoadjuvant therapy was associated with complete pathologic response or significant downstaging of disease, there was a trend for improved survival in the downstaged group, but it did not reach statistical significance (P = 0.14). Third, adjuvant therapy was associated with improved 5-year survival (P = 0.03), particularly for combination chemotherapy and radiotherapy (P = 0.02). PMID- 16351030 TI - Effect of N-acetyl cysteine on Helicobacter pylori. AB - BACKGROUND: Use of mucolytic agents that result in reduced mucous viscosity of the gastric mucous has been suggested to have an additive effect in curing Helicobacter pylori infection. METHODS: Seventy Hpylori-positive patients were given either eradication treatment consisting of 500 mg clarithromycin bid and 30 mg lansoprazole bid for 10 days plus 10 mL (400 mg) N-acetyl cysteine (NAC) liquid tid (AC group) or eradication treatment only (control group). The results were compared 1 month after the completion of the treatment. RESULTS: Fifty-eight patients were available for statistical analysis. Of the 28 patients in the AC group, 14 (50.0%) eradicated the infection after treatment, whereas only 7 of 30 (23.3%) patients in the control group had negative results. The difference between the AC group and the control group was statistically significant (P = 0.034). In both groups, there was no difference in the number of smokers and in the eradication rates between smokers and nonsmokers. Eradication treatment with or without NAC caused no significant side effects in either group. CONCLUSIONS: Our findings suggest that NAC has an additive effect on the eradication rates of H pylori obtained with dual therapy with lansoprazole and clarithromycin. NAC does not have any known activity against H pylori, but it may improve the delivery of antibiotics at the site of infection due to its ability to reduce the thickness of the mucus. PMID- 16351031 TI - Psychosomatic pain: new insights and management strategies. AB - At least 40 to 60 percent of women and at least 20 percent of men with chronic pain disorders report a history of being abused during childhood and/or adulthood. This incidence of abuse is two to four times higher than in the general population. Patients with more severe or frequent abuse, usually during childhood and worse if sexual in nature. often develop specific syndromes or combinations of syndromes. These syndromes include posttraumatic stress disorder, fibromyalgia, and other conditions characterized by repression, somatization, and increased utilization of medical care. Psychosomatic symptoms and dysfunctional behaviors may emerge as these patients seek attention and validation of their suffering, while paradoxically repressing painful memories of trauma. Behavioral observations and key features of the physical examination may greatly help the clinician identify both the presence and severity of psychosomatic disease. In addition, it is very interesting that various studies document physiologic changes in the brains of patients with a history of abuse and in patients with a diagnosis of fibromyalgia. These studies suggest that abuse may physiologically and developmentally increase a person's susceptibility to pain and that some organic changes may be associated with psychogenic disease. Diagnosis and treatment of even the most challenging patients with chronic pain is much more effective if it includes (a) careful inquiry about any history of past or present abuse or other severe trauma, (b) empathy and constructive validation of disease and suffering, (c) recognition of dysfunctional pain behaviors and personality traits, (d) documentation of nonanatomic as well as anatomic features on examination, (e) multidisciplinary treatments including psychotherapy whenever indicated, and (f) noninvasive procedures and alternatives to potentially habit forming medications whenever possible and appropriate. Furthermore, it has been shown that helping patients gain insight about the relationship between abuse and their current symptoms leads to decreased health care utilization. Practical guidelines are provided for identifying psychopathology, communicating effectively, and achieving better treatment outcomes for these unfortunate patients. PMID- 16351032 TI - Acetaminophen overdose in pregnancy. AB - Acetaminophen (APAP) is the most common drug overdose in pregnancy. Available data regarding APAP overdose in pregnancy is limited to case reports and a small prospective case series. APAP has been demonstrated to cross the placenta and in toxic doses may harm the fetal and maternal hepatocytes. Fetal hepatocytes metabolize APAP into both active and toxic metabolites. These toxic metabolites may cause fetal hepatic necrosis. N-acetylcysteine (NAC) has also been demonstrated to cross the placenta and may bind toxic metabolites in both the mother and the fetus. Limited data suggest that the majority of morbidity and mortality from APAP overdose can be averted by initiation of NAC within the first 16 hours of ingestion and possibly even later. NAC may be safely administered during pregnancy and should be initiated early after APAP overdosage. The literature was reviewed through the use of OvidMEDLINE database, encompassing 1966 to the present. Searches were conducted using the key words acetaminophen, paracetamol, N-acetylcysteine, overdose, and hepatotoxicity. The search was further refined by selecting articles that contained these search words together with the key word pregnancy. Only English language papers were reviewed. Articles were selected on the basis of relevance to the topic. Pertinent citations found in the selected articles were also reviewed. PMID- 16351033 TI - A comprehensive review of disseminated Salmonella arizona infection with an illustrative case presentation. AB - Salmonella arizona is known to cause infection in reptiles and other animals. Disseminated human infection is rare, except in the setting of a deficient immune system. The following is a unique account of disseminated infection including pericardial involvement. Unusual features include nonreptile vector transmission and eastern seaboard (rather than southwestern) locale. A comprehensive literature review of disseminated S arizona infections is presented describing the types of infection, sources of exposure, underlying conditions, locale, treatments, and outcomes. PMID- 16351034 TI - Methicillin-resistant Staphylococcus aureus empyema necessitatis in an eight month-old child. AB - Empyema necessitatis is a rare complication of empyema characterized by a spontaneous extension of pus from the pleural space into adjacent soft tissues. It is uncommon in the pediatric population and is usually caused by Mycobacterium tuberculosis. This report describes the youngest reported case of empyema necessitatis. The clinical examination and CT scan are diagnostic. The causative organism was methicillin-resistant Staphylococcus aureus, which has not been previously reported. Treatment involves drainage of the abscess and appropriate antimicrobial therapy. PMID- 16351035 TI - Klippel-Feil syndrome associated with pneumatocyst of the right cervical rib. AB - A patient with Klippel-Feil syndrome reported with a pneumatocyst of the proximal epiphysis of a right cervical rib. There was no additional skeletal abnormality except for fusion of the C3 and C4 vertebral bodies. To the best of our knowledge, this is the first reported instance of this particular association. PMID- 16351036 TI - A case of allergic bronchopulmonary aspergillosis leading to pneumonia with unusual organisms. AB - We describe the case of a 50-year-old male with a history of asthma and seizure disorder who presented with a 5-month history of dyspnea. The patient had been treated with multiple courses of antibiotics for presumed community-acquired pneumonia before being determined to have allergic bronchopulmonary aspergillosis (ABPA) by serologic and radiographic criteria. Inflammation resulting from this disease had potentiated a postobstructive pneumonia caused by Nocardia asteroides and Stenotrophomonas maltophilia. Therapy with corticosteroids, trimethoprim sulfa, and voriconazole failed to prevent subsequent destruction of the right upper lobe and the patient required surgical intervention. The discussion emphasizes the diagnostic criteria for ABPA including historic, serologic, and radiographic findings; staging, and treatment. Other possible diagnoses, such as invasive pulmonary aspergillosis, chronic necrotizing aspergillosis, and hyper IgE syndrome are also briefly reviewed. PMID- 16351037 TI - Unusual case of pyopneumothorax in Tennessee. AB - Rupture of a coccidioidal pulmonary cavity with subsequent pyopneumothorax is a rare clinical event, even in areas endemic for coccidioidomycosis. Our encounter with a patient diagnosed with this condition in northeast Tennessee serves notice to clinicians that coccidioidomycosis is indeed a traveling fungal disease, and practitioners must be alert to common and uncommon manifestations of infection associated with this fungus. A literature review pertaining to coccidioidal pyopneumothorax revealed that patients usually present with a recent onset of chest pain. Serologic testing and pleural fluid culture are highly useful, and management includes surgical intervention with or without antifungal therapy. PMID- 16351038 TI - Disseminated cat scratch disease with vertebral osteomyelitis and epidural abscess. AB - A 5-year-old boy with cat scratch disease presented with fever of unknown origin and osteomyelitis of the thoracic spine and epidural abscess. He did not have localizing signs or symptoms. Computed tomography of the abdomen, which was initially negative, showed hepatosplenic disease. Cat scratch disease has variable systemic presentations and should be included in the differential diagnosis of fever of unknown origin if an epidemiologic risk factor is present. PMID- 16351039 TI - Isolated acetabular osteoporosis in TOH in pregnancy: a case report. AB - Transient osteoporosis of the hip (TOH) is a rare, self-limiting syndrome characterized by hip pain in the 2nd or 3rd trimester of pregnancy. Conventional radiography shows demineralization of the femoral head. Diagnosis is confirmed by MRI scan. Over the course of a few months, the radiological abnormalities and the pain disappear spontaneously with complete resolution. We report the case of a patient with isolated acetabular involvement during pregnancy, confirmed by MRI scan, who obtained spontaneous remission of her symptoms. PMID- 16351040 TI - Acute progressive bilateral carpal tunnel syndrome after upper respiratory tract infection. AB - This report describes the case of a 32-year-old male presenting with acute progressive bilateral carpal tunnel syndrome after a benign upper respiratory tract infection. Serial nerve conduction studies confirmed progressive entrapment of the median nerves in the carpal tunnel to the point of axonal damage. Surgical decompression relieved the entrapment, and nerve conduction studies improved. PMID- 16351041 TI - Does olanzapine-fluoxetine combination increase the risk of mania in poorly compliant bipolar depressed patients? PMID- 16351042 TI - Percutaneous intervention of left main coronary artery disease: a fresh look at a class III recommendation. PMID- 16351043 TI - Patient's page. Bone health facts. PMID- 16351045 TI - Advances in the total synthesis of biologically important marine macrolides. PMID- 16351046 TI - Total synthesis of marine polycyclic ethers. PMID- 16351047 TI - Total synthesis of oxacyclic macrodiolide natural products. PMID- 16351048 TI - Convergent strategies for syntheses of trans-fused polycyclic ethers. PMID- 16351049 TI - Nonanomeric spiroketals in natural products: structures, sources, and synthetic strategies. PMID- 16351050 TI - Recent progress of the synthetic studies of biologically active marine cyclic peptides and depsipeptides. PMID- 16351051 TI - Synthetic chemistry of halichlorine and the pinnaic acids. PMID- 16351052 TI - Syntheses of natural products having an epoxyquinone structure. PMID- 16351053 TI - Total synthesis of natural tert-alkylamino hydroxy carboxylic acids. PMID- 16351054 TI - Total synthesis of alpha-pyrone meroterpenoids, novel bioactive microbial metabolites. PMID- 16351055 TI - Progress on the chemistry of dibenzocyclooctadiene lignans. PMID- 16351056 TI - Syntheses around the transglycosylation step in peptidoglycan biosynthesis. PMID- 16351057 TI - Total synthesis of ingenol. PMID- 16351058 TI - Reserpine: a challenge for total synthesis of natural products. PMID- 16351059 TI - Total synthesis of selected bioactive natural products: illustration of strategy and design. PMID- 16351060 TI - A case study in biomimetic total synthesis: polyolefin carbocyclizations to terpenes and steroids. PMID- 16351061 TI - Biosynthetic and biomimetic electrocyclizations. PMID- 16351062 TI - Recent advances in natural product synthesis by using intramolecular Diels-Alder reactions. PMID- 16351063 TI - Pore formation coupled to ion transport through lipid membranes as induced by transmembrane ionic charge imbalance: atomistic molecular dynamics study. AB - We have employed atomic-scale molecular dynamics simulations to address ion transport through transient water pores in phospholipid membranes. The formation of a water pore is induced by a transmembrane ionic charge imbalance, which gives rise to a significant potential difference across the membrane. The subsequent transport of ions through the pore discharges the transmembrane potential and makes the water pore metastable, leading eventually to its sealing. The findings highlight the importance of ionic charge fluctuations in spontaneous pore formation and their role in ion leakage through protein-free lipid membranes. PMID- 16351064 TI - MRI thermometry based on PARACEST agents. AB - A novel magnetic resonance imaging (MRI) thermometry technique is demonstrated in vitro based upon the use of a PARACEST (PARAmagnetic Chemical Exchange Saturation Transfer) agent. This new method takes advantage of the high concentration of bulk water (the readout signal for imaging) and the hyperfine frequency shift properties of PARACEST agents. For two prototypes, Dy(1)3+ and Eu(2)-, the chemical shifts (delta, in ppm) of the Ln3+-bound water molecules are linearly dependent on temperature (T, in degrees C) over the range of 20-50 degrees C (delta = 6.9 x T - 944.7 and delta = -0.4 x T + 64.6, respectively). This offers the exciting possibility of improving the temperature dependencies approximately 690- and approximately 40-fold over the most widely used water PRF thermometry (Proton Resonance Frequency: -0.01 ppm/ degrees C). PMID- 16351065 TI - Ultrafast intermolecular zero quantum spectroscopy. AB - Clinical magnetic resonance spectroscopy is typically limited by magnetic inhomogeneities which destroy spectral resolution, but intermolecular zero quantum coherences (iZQCs) are insensitive to such inhomogeneities. iZQC resolution in vivo, however, has been hampered by physiological fluctuations over the time scale of the two-dimensional acquisition. A faster iZQC sequence will allow us to average away these fluctuations, and thus we present a new approach to ultrafast two-dimensional spectroscopy. This communication reports iZQC experiments acquiring up to 31 t1-points per scan, as well as extensions to a broad range of other 2D sequences. PMID- 16351066 TI - Combinatorial biomimetics. optimization of a composition of copper(II) poly-L histidine complex as an electrocatalyst for O2 reduction by scanning electrochemical microscopy. AB - A simple approach to prepare and characterize biomaterial-based electrocatalysts for oxygen reduction was carried out. Poly-l-histidine was used as a matrix and ligand to complex Cu2+ to mimic the active sites of laccases. A modified glassy carbon (GC) electrode with Cu2+-poly-l-histidine complex decreases the oxygen reduction overpotential as compared with the bare GC electrode. An array of Cu2+ poly-l-histidine spots with different compositions was deposited on a GC substrate, and their catalytic activity for oxygen reduction was evaluated by a scanning electrochemical microscopy-based screening technique. The electrocatalytic activities of complexes for oxygen reduction strongly depended on the mole ratio of Cu2+ to poly-l-histidine and the applied potential of the substrate. PMID- 16351067 TI - Supramolecular oxidation of anilines using hydrogen peroxide as stoichiometric oxidant. AB - 6A,6D-Di-O-(propan-2-on-1,3-diyl) alpha-cyclodextrin-6A,6D-dicarboxylate (2alpha) and 6A,6D-di-O-(propan-2-on-1,3-diyl) beta-cyclodextrin-6A,6D-dicarboxylate (2beta) were found to catalyze the oxidation of aromatic amines in the presence of hydrogen peroxide. The products were the corresponding nitro compounds or in some cases azo-, azoxy-, or other dimerization products. The catalysis was found to follow enzyme kinetics giving a rate increase (kcat/kuncat) of up to 1100 in the best case. PMID- 16351068 TI - A polydiacetylene-based fluorescent sensor chip. AB - Self-assembled diacetylene vesicles were spotted and immobilized on aldehyde modified glass substrates using conventional microarray technology. Irradiation of the immobilized diacetylenes allowed generation of nonfluorescent "blue-phase" polydiacetylene (PDA) arrays. Specific interaction of the PDA vesicle arrays with carbohydrates or poly(acrylic acid) solutions afforded fluorescent profiles. PMID- 16351069 TI - The ferrous verdoheme-heme oxygenase complex is six-coordinate and low-spin. AB - A biosynthetic and enzymatic method was developed for the preparation of 13C labeled verdoheme, which permits the 13C NMR spectroscopic characterization of this elusive intermediate in the heme oxidation path catalyzed by the enzyme heme oxygenase. The 13C NMR data indicate that the ferrous verdoheme complex of Neisseria meningitides heme oxygenase is hexacoordinate and diamagnetic, with a proximal histidine and likely a distal hydroxide as axial ligands. The coordination number and spin state of the ferrous verdoheme-heme oxygenase complex is in stark contrast to the pentacoordinate and paramagnetic nature of the heme-heme oxygenase complex and heme centers in general. PMID- 16351070 TI - Cellular logic with orthogonal ribosomes. AB - The creation and use of unnatural molecules to control cellular function is a long standing goal of the chemical community, but in general, these efforts have been directed at finding molecules to inhibit or activate a particular molecular target or function, or to elicit a particular phenotype. Here we show that multiple unnatural molecules (orthogonal ribosomes) can be used combinatorially, in a single cell, to program Boolean logic functions. These experiments show how attention to the molecular specificity of noncovalent interactions between unnatural macromolecules allows the synthesis of complex function from the "bottom-up" in living matter. PMID- 16351071 TI - An alternative of CdSe nanocrystal emitters: pure and tunable impurity emissions in ZnSe nanocrystals. AB - The concept, decoupling doping from nucleation and/or growth, allows us to dope nearly all nanocrystals in a given sample which is indicated by complete quenching of the host emission and bright emission from the dopants at characteristic wavelengths tunable in most parts of the visible window using a ZnSe host. In an extreme case, ZnSe coated MnSe nanocrystals (MnSe:ZnSe) emit similarly as commonly known doped nanocrystals. In comparison with CdSe nanocrystals, these alternative emitters not only are intrinsically less toxic but also show some unexpected and expected advantages: stable against thermal and environmental changes, zero reabsorption, and no Forster energy transfer. In addition to their applications to replace CdSe based nanocrystal emitters, the unique structure and properties of the doped nanocrystals are of interest for studying fundamental issues in the field. PMID- 16351072 TI - RNA GG x UU motif binds K+ but not Mg2+. AB - A hairpin model of the group I intron P5b loop was synthesized with [8-13C-7-15N] guanosine in the GG.UU metal binding site, [7-15N]-guanosine at a nonbinding site, and [3-15N]-uridine. 15N NMR showed specific binding for Co(NH3)63+ and K+, but not for Zn2+, Cd2+, or Mg2+. PMID- 16351073 TI - Self-assembly of DNA double-double crossover complexes into high-density, doubly connected, planar structures. AB - We designed a molecular complex, the double-double crossover, consisting of four DNA double helices connected by six reciprocal exchanges. Atomic force micrographs suggest that double-double crossover complexes self-assemble into high-density, doubly connected, two-dimensional, planar structures. Such structures may be suitable as substrates for the deposition of nanomaterials in the creation of high-density electrical and quantum devices. We speculate about a modified double-double crossover complex that might self-assemble into high density, doubly connected, three-dimensional structures. PMID- 16351074 TI - Synthesis, neutron structure, and reactivity of the bis(dihydrogen) complex RuH2(eta(2)-H2)2(PCyp3)2 stabilized by two tricyclopentylphosphines. AB - Treatment of Ru(eta4-C8H12)(eta6-C8H10) with 3 bar H2 in the presence of 2 equiv of tricyclopentylphosphine (PCyp3) in pentane resulted in the isolation of the new bis(dihydrogen) complex RuH2(eta2-H2)2(PCyp3)2 (2), characterized by NMR and single-crystal X-ray and neutron diffraction. The single-crystal neutron diffraction study is the first carried out for a bis(dihydrogen) complex. The coordination geometry around the metal center is a distorted octahedron defined by the two phosphines in a trans configuration (making an angle of 168.9(1) degrees ), two cis dihydrogen ligands, and two hydrides trans to them, defining the equatorial plane. The H-H bond distances (0.825(8) and 0.835(8) A) are characteristic of two "unstretched" dihydrogen ligands. H/D exchange between the Ru-H and the C-D bonds of deuterated benzene is observed within 1 h, leading to the formation of various isotopomers RuHxD6-x(PCyp3)2 (with x = 0-6). 2 is a catalyst precursor for ethylene coupling (20 bar, 293 K) to a functionalized arene (Murai reaction). We found a 90% conversion of acetophenone to 2 ethylacetophenone within 35 min, whereas 10 h was needed in the same conditions using the analogous tricyclohexylphosphine complex, RuH2(eta2-H2)2(PCy3)2, the best catalyst precursor, at room temperature, prior to this work. PMID- 16351075 TI - Characterization of kinetics and products of the Baeyer-Villiger oxygenase MtmOIV, the key enzyme of the biosynthetic pathway toward the natural product anticancer drug mithramycin from Streptomyces argillaceus. AB - MtmOIV, the key oxygenase of the mithramycin biosynthetic pathway in Streptomyces argillaceus, was proven to act initially as Baeyer-Villiger monooxygenase, but may also catalyze various follow-up reaction steps. The reaction of the overexpressed pure His6-tagged enzyme with its substrate premithramycin B was studied. Various intermediates and products were isolated and physicochemically characterized, several of them being previously unknown compounds. This is the first example in which a bacterial enzyme was unequivocally proven to act as Baeyer-Villigerase with its natural substrate, that is, in its natural context. PMID- 16351076 TI - Structural control of self-assembled nanofibers by artificial beta-sheet peptides composed of D- or L-isomer. AB - A novel method for the control of peptide self-assembly has been developed by using synthetic triblock-type beta-sheet peptides composed of l- or d-amino acid, 1L and 1D, as building blocks. The peptides 1L and 1D self-assemble into beta sheet nanofibers with left- and right-handed twists, respectively, under appropriate condition. On the other hand, the 1L/1D binary mixture was found to form only globular aggregates at the same condition. Thus, amyloid-like nanofiber formation and its nanostructure could be successfully regulated by the stereospecificity of the constituent peptide species. PMID- 16351077 TI - A mixed-valence coordination polymer featuring two-dimensional ferroelectric order: {[Cu(I)4Cu(II)(Et2dtc)2Cl3][Cu(II)(Et2dtc)2]2(FeCl4)}n (Et2dtc- = diethyldithiocarbamate). AB - The first mixed-valence coordination polymer indicating ferroelectric properties, {[CuI4CuII(Et2dtc)2Cl3][CuII(Et2dtc)2]2(FeCl4)}n (Et2dtc- = diethyldithiocarbamate), has been synthesized and crystallographically characterized. This complex has a two-dimensional infinite square lattice structure, including both Cu(I) and Cu(II) ions, which is confirmed by XPS and SQUID measurements. In addition, the ferroelectric behaviors are revealed by the dielectric measurements with a LCR meter and a ferroelectric tester. PMID- 16351078 TI - Deposition of CTAB-terminated nanorods on bacteria to form highly conducting hybrid systems. AB - Gold nanorods and nanospheres capped with positively charged CTAB are deposited on the surface of gram-positive bacterium having negatively charged teichoic acid brushes. The deposition rate is more than an order of magnitude faster than that for peptide and nucleic acid capped nanoparticles. For the nanorods, never been reported before, the strong electrostatic attraction causes the rods to bend conformally over the curved bacterium surface in random orientations. This leads to formation of an efficient percolating cluster (i.e., low contact resistance and high radius of gyration) with 104-fold higher current at 3-fold lower coverage compared to a monolayer from nanospheres. The high monolayer conductivity at only approximately 10% coverage for nanorods has potential for fabricating a novel class of bioelectronic devices by coupling electronic "nanocircuitry" at the surface to the biological function of microorganisms. PMID- 16351079 TI - A propeller-like uranyl metallomesogen. AB - Uranyl triflate forms with three imidazo[4,5-f]-1,10-phenanthroline ligands a propeller-like complex that exhibits a hexagonal columnar phase. The ligand is not liquid-crystalline, but a mesophase is induced upon complex formation with the uranyl salt. The thermal behavior has been investigated by polarizing optical microscopy and by high-temperature X-ray diffraction. A model of the stacking of the molecules in the mesophase is proposed. PMID- 16351080 TI - Bright fluorescent nanodiamonds: no photobleaching and low cytotoxicity. AB - Diamond nanocrystals emit bright fluorescence at 600-800 nm after irradiation by a 3 MeV proton beam (5 x 1015 ions/cm2) and annealing at 800 degrees C (2 h) in vacuum. The irradiation/annealing process yields high concentrations of nitrogen vacancy defect centers ( approximately 107 centers/mum3), making possible visualization of the individual 100 nm diamond crystallites using a fluorescence microscope. The fluorescent nanodiamonds (FND) show no sign of photobleaching and can be taken up by mammalian cells with minimal cytotoxicity. The nanomaterial can have far-reaching biological applications. PMID- 16351081 TI - Self-assembling a molecular pegboard. AB - This paper describes the design, construction, and application of a self assembled, chemically addressable DNA nanogrid composed of DNA tiles of nanometer dimensions. This self-assembled structure permits precise placement of molecules at predetermined locations on a "molecular pegboard". We used the indexed DNA nanogrids to identify single molecules of DNA that hybridize at particular locations. PMID- 16351082 TI - Simultaneous, segregated storage of two agents in a multicompartment micelle. AB - The simultaneous, segregated storage of two different chromophores in a multicompartment micelle, which is formed from self-assembly of (polyethylethylene)(polyethylene oxide)(polyperfluoropropylene oxide) mikto-arm star terpolymers in water, was investigated by spectrophotometry. The multicompartment micelles, with segregated micellar cores composed of fluorocarbon and hydrocarbon compartments, can simultaneously absorb two chemically different molecules. These two molecules were confined into their preferred compartments with high selectivity. PMID- 16351083 TI - Creating responsive surfaces with tailored wettability switching kinetics and reconstruction reversibility. AB - We report on the formation of responsive surfaces with tailorable surface reconstruction kinetics and switching hysteresis by thiolene radical addition of mercaptoalkanols with variable lengths to poly(vinylmethylsiloxane) networks. Exposing these elastomer surfaces to water results in rearrangement of the hydrophilic alkanes at the surface. The rearrangement kinetics decreases with increasing number of the methylene spacers (n) in the mercaptoalkanol. The response kinetics is found to be very fast for n = 2 and 6. For instance, upon exposing to water, the water contact angle on 3-mercaptopropanol-based surfaces decreases by approximately 35 degrees at the rate of 2 degrees /s. The high flexibility of the siloxane backbone endows these materials with switching longevity; the materials were able to switch their wettability over 10 cycles with minimum hysteresis. Increasing the number of methylene spacers to n = 11 decreases the surface reorganization dramatically. Formation of semicrystalline regions in such materials (detected via IR) is responsible for initial "sluggish" kinetics and eventual surface "freezing". PMID- 16351084 TI - Acceleration of a nanomotor: electronic control of the rotary speed of a light driven molecular rotor. AB - A new second-generation light-driven molecular motor was designed, in which the presence of a potential electronic push-pull system leads to a significant increase of the rate of rotation compared to previous motor systems, without disturbing its overall unidirectionality. PMID- 16351085 TI - Rhodium-catalyzed alkyne hydrothiolation with aromatic and aliphatic thiols. AB - Alkyne hydrothiolation is a potentially attractive method for the formation of vinyl sulfides, which are valuable synthetic intermediates. Known methods for hydrothiolation using alkyl thiols are quite limited. We report herein that Tp*Rh(PPh3)2 (Tp* = hydrotris(3,5-dimethylpyrazolyl)borate) is a highly active catalyst for alkyne hydrothiolation with alkyl and aryl thiols. Hydrothiolation using alkyl thiols proceeds with excellent regioselectivity, providing convenient access to branched alkyl vinyl sulfides, which are difficult to synthesize by other means. A mixture of regioisomers is obtained when using aryl thiols, with the branched isomer as the major product, opposite that reported for other Rh complexes. PMID- 16351086 TI - Living ring-opening polymerization of N-sulfonylaziridines: synthesis of high molecular weight linear polyamines. AB - Sulfonylaziridines have been identified as excellent monomers for living ring opening polymerization initiated by nucleophilic sulfonylamides. The resulting polymers exhibit low polydispersities and controllable molecular weights. The enantiopurity of the monomer plays a key role: racemic samples yield soluble polymers of target molecular weights, while enantiopure samples produce insoluble polymers with molecular weights significantly below theoretical values. Dynamic light scattering and kinetics of polymerization are discussed. PMID- 16351087 TI - A multifunctional Pasteurella multocida sialyltransferase: a powerful tool for the synthesis of sialoside libraries. AB - A multifunctional sialyltransferase has been cloned from Pasteurella multocida strain P-1059 and expressed in E. coli as a truncated C-terminal His6-tagged recombinant protein (tPm0188Ph). Biochemical studies indicate that the obtained protein is (1) an alpha2,3-sialyltransferase (main function), (2) an alpha2,6 sialyltransferase, (3) an alpha2,3-sialidase, and (4) an alpha2,3-trans sialidase. The recombinant tPm0188Ph is a powerful tool in the synthesis of structurally diverse sialoside libraries due to its relaxed substrate specificity, high solubility, high expression level, and multifunctionality. PMID- 16351088 TI - Chemoenzymatic dynamic kinetic resolution of primary amines. AB - An efficient process for dynamic kinetic resolution of amines was developed by combining a ruthenium-catalyzed racemization with a lipase-catalyzed resolution. A variety of unfunctionalized primary amines were transformed into one enantiomer of the amide in high yield and high enantioselectivity. PMID- 16351089 TI - Catalytic enantioselective aza-Henry reaction with broad substrate scope. AB - In situ generated azomethines from readily available precursors react with nitromethane in the presence of 120 mol % of CsOH.H2O and 12 mol % of quinine- and cinchonidine-derived quaternary ammonium chlorides to provide the corresponding aza-Henry adducts in good yields and very high selectivities. It represents the first general enantioselective aza-Henry method for azomethines derived from enolizable aldehydes, giving rise to enantiomeric excesses above 94%. In addition, the reactions with nitroethane afforded high diastereo- and enantioselectivities (syn:anti up to 95:5; up to 98% ee for syn). PMID- 16351090 TI - Disubstituted imidazolium-2-carboxylates as efficient precursors to N heterocyclic carbene complexes of Rh, Ru, Ir, and Pd. AB - N,N'-Disubstituted imidazolium-2-carboxylates are efficient precursors to NHC complexes of Rh, Ir, Pd, and Ru. PMID- 16351091 TI - Mass spectrometry assisted assignment of NMR resonances in reductively 13C methylated proteins. AB - Reductive 13C-methylation of proteins has been used as an isotope labeling strategy to study protein structure, function, and dynamics by nuclear magnetic resonance (NMR) spectroscopy. However, assigning the resulting 13C-dimethylamine peaks in a 1H-13C NMR spectrum has proved to be difficult, but it is important to expand the scope of the method. The assignment strategy presented here utilizes mass spectrometry (MS) for sequence identification and varying 13C/12C isotope ratios to correlate with NMR data. The site-specific reactivity of the lysines and N-terminal amine of a protein is exploited to produce a sample with varying 13C/12C ratios at each dimethylamine. MS and NMR are used to quantitate and correlate these ratios in order to assign peaks in the 1H-13C NMR spectrum. Hen egg white lysozyme was used as a model protein to demonstrate this assignment strategy. PMID- 16351092 TI - Identification and hydrogenation of C2 on Pt(111). AB - Reflection absorption infrared spectroscopy (RAIRS) and temperature-programmed desorption (TPD) were used to identify the molecular species formed upon the reaction of hydrogen with surface carbon that is deposited by exposing acetylene to a Pt(111) surface held at 750 K. At this temperature, the acetylene is completely dehydrogenated and all hydrogen is desorbed from the surface. Upon subsequent hydrogen exposure at 85 K followed by sequential annealing to higher temperatures, ethylidyne (CCH3), ethynyl (CCH), and methylidyne (CH) are formed. The observation of these species indicates that carbon atoms and C2 molecules exist as stable species on the surface over a wide range of temperatures. Through a combination of RAIRS intensities, hydrogen TPD peak areas, and Auger electron spectroscopy, quantitative estimates of the coverages of the various species were obtained. It was found that 79% of the acetylene-derived carbon was in the form of C2 molecules, with the remainder in the form of carbon atoms. Essentially all of the acetylene-derived carbon could be hydrogenated. In contrast, 85% of an equivalent coverage of carbon deposited by ethylene exposure at 750 K was found to be inert toward hydrogenation. PMID- 16351093 TI - Spectroscopic characterization of hydroxide and aqua complexes of Fe(II) protoheme, structural models for the axial coordination of the atypical heme of membrane cytochrome b6f complexes. AB - Electronic absorption and resonance Raman (RR) spectra are reported for hydroxide and aqua complexes of iron(II)-protoporphyrin IX (Fe(II)PP) respectively formed in alkaline and neutral aqueous solutions. These compounds with weak axial ligand(s) represent a biomimetic approach of the unusual coordination of the atypical heme c(i) of membrane cytochrome b6f complexes. Absorption spectra and spectrophotometric titrations show that Fe(II)PP in alkaline aqueous cetyltrimethylammonium bromide (CTABr) binds one hydroxide ion, forming a five coordinated high-spin (HS) complex. In alkaline aqueous ethanol, we confirm the formation of a dihydroxy complex of Fe(II)PP. In the RR spectra of Fe(II)PP dissolved in neutral aqueous CTABr, a mixture of a four-coordinated intermediate spin form with an HS monoaqua complex (Fe(II)PP(H2O)) was observed. The spectroscopic information obtained for Fe(II)PP(OH-), Fe(II)PP(H2O), and Fe(II)PP(OH-)2 was compared with that previously reported for the 2 methylimidazole and 2-methylimidazolate complexes of Fe(II)PP, representative of the most common axial ligation in HS heme proteins. This investigation reveals a very remarkable analogy in the spectral properties of, in one hand, the Fe(II)PP(H2O) and mono-2-methylimidazole complexes and, in the other hand, the Fe(II)PP(OH-) and mono-2-methylimidazolate complexes. The comparisons of the absorption and RR spectra of Fe(II)PP(OH-) and Fe(II)PP(OH-)2 clearly establish that both a redshift of the pi-pi electronic transitions and an upshift of the v8 RR frequency are spectral parameters indicative of porphyrin doming in HS ferrous complexes. Based upon isotopic substitutions (16OH-,16OD-, and 18OH-), stretching modes of the Fe-OH bond(s) of a ferrous porphyrin were assigned for the first time, i.e., at 435 cm(-1) for Fe(II)PP(OH-) (nu(Fe(II)-OH-)) and at 421 cm(-1) for Fe(II)PP(OH-)2 (nu(s)(Fe(II)-(OH-)2). The spectroscopic and redox properties of Fe(II)PP(H2O), Fe(II)PP(OH-), and heme c(i) were discussed and favor a water coordination for the heme c(i) iron. PMID- 16351094 TI - Alkyne hydrosilylation catalyzed by a cationic ruthenium complex: efficient and general trans addition. AB - The complex [Cp*Ru(MeCN)3]PF6 is shown to catalyze the hydrosilylation of a wide range of alkynes. Terminal alkynes afford access to alpha-vinylsilane products with good regioselectivity. Deuterium labeling studies indicate a clean trans addition process is at work. The same complex is active in internal alkyne hydrosilylation, where absolute selectivity for the trans addition process is maintained. Several internal alkyne substrate classes, including propargylic alcohols and alpha,beta-alkynyl carbonyl compounds, allow regioselective vinylsilane formation. The tolerance of a wide range of silanes is noteworthy, including alkyl-, aryl-, alkoxy-, and halosilanes. This advantage is demonstrated in the direct synthesis of triene substrates for silicon-tethered intramolecular Diels-Alder cycloadditions. PMID- 16351095 TI - Guanidinophosphazenes: design, synthesis, and basicity in THF and in the gas phase. AB - A principle for creating a new generation of nonionic superbases is presented. It is based on attachment of tetraalkylguanidino, 1,3-dimethylimidazolidine-2-imino, or bis(tetraalkylguanidino)carbimino groups to the phosphorus atom of the iminophosphorane group using tetramethylguanidine or easily available 1,3 dimethylimidazolidine-2-imine. Seven new nonionic superbasic phosphazene bases, tetramethylguanidino-substituted at the P atom, have been synthesized. Their base strengths are established in tetrahydrofuran (THF) solution by means of spectrophotometric titration and compared with those of eight reference superbases designed specially for this study, P2- and P4-iminophosphoranes. The gas-phase basicities of several guanidino- and N',N',N'',N''-tetramethylguanidino (tmg)-substituted phosphazenes and their cyclic analogues are calculated, and the crystal structures of (tmg)3P=N-t-Bu and (tmg)3P=N-t-Bu x HBF4 are determined. The enormous basicity-increasing effect of this principle is experimentally verified for the tetramethylguanidino groups in the THF medium: the basicity increase when moving from (dma)3P=N-t-Bu (pKalpha = 18.9) to (tmg)3P=N-t-Bu (pKalpha = 29.1) is 10 orders of magnitude. A significantly larger basicity increase (up to 20 powers of 10) is expected (based on the high-level density functional theory calculations) to accompany the similar gas-phase transfer between the (dma)3P=NH and (tmg)3P=NH bases. Far stronger basicities still are expected when, in the latter two compounds, all three dimethylamino (or tetramethylguanidino) fragments are replaced by methylated triguanide fragments, (tmg)2C=N-. The gas-phase basicity (around 300-310 kcal/mol) of the resulting base, [(tmg)2C=N-]3P=NH, having only one phosphorus atom, is predicted to exceed the basicity of (dma)3P=NH by more than 40 powers of 10 and to surpass also the basicity of the widely used commercial [(dma)3P=N]3P=N-t-Bu (t-BuP4) superbase. PMID- 16351096 TI - Hybrids of the hemiasterlin analogue taltobulin and the dolastatins are potent antimicrotubule agents. AB - The targeting of microtubules is an important mechanism for cancer chemotherapy. However, there is still a need for improved antimicrotubule agents. A number of seemingly structurally disparate peptidic natural products inhibit tubulin polymerization by binding to a region of the tubulin heterodimer close to the vinca binding site. An analogue of the naturally occurring tripeptide hemiasterlin, taltobulin (HTI-286, 3), has advanced to clinical trials. Structure activity relationship studies of 3 have revealed critical structural elements necessary for antimicrotubule activity that correspond to comparable groups in the amino terminus tripeptide region of the dolastatins. To investigate the structural relationship between the hemiasterlins and the more complex dolastatins, hybrid compounds composed of 3 and the carboxy terminus dipeptides of dolastatin 10, or the dolastatin 15 analogue cemadotin, were synthesized. The resulting hybrid compounds were potent antimicrotubule agents, thus establishing a structural relationship between the hemiasterlins and the dolastatins. This relationship may be useful in the design of analogues having improved activity in resistant cell lines expressing the P-glycoprotein transporter, for establishing structural relationships with other classes of peptidic antimicrotubule agents, or for modeling studies of the tubulin binding site of these agents. PMID- 16351097 TI - Synthesis of chiral self-assembling rhombs and their characterization in solution, in the gas phase, and at the liquid-solid interface. AB - Chiral, enantiopure metallo-supramolecular rhombs self-assemble in solution through coordination of bis-pyridyl-substituted ligands with (en)M(NO3)2 (en = ethylenediamine, M = Pd(II), Pt(II)). Characterization by NMR and CD spectroscopy in solution and by ESI-FT-ICR mass spectrometry in the gas phase suggests that an equilibrium exists in water/methanol of a major 2:2 complex and a minor 3:3 complex of ligands and metal corners. In the gas phase, doubly charged 2:2 complexes fragment into two identical singly charged halves followed by metal mediated C-H and C-C bond activation reactions within the ethylenediamine ligands. Electrochemical scanning tunneling microscopy (EC-STM) provides in situ imaging of the complexes even with submolecular resolution. Flat-lying rhombs are deposited under potential control from an aqueous electrolyte on a Cu(100) electrode surface precovered by a tetragonal pattern of chloride anions from the supporting electrolyte. Chirality induces the formation of only one domain orientation. Density functional calculations help to interpret the STM images. PMID- 16351099 TI - Application of MCD spectroscopy and TD-DFT to a highly non-planar porphyrinoid ring system. New insights on red-shifted porphyrinoid spectral bands. AB - The first magnetic circular dichroism (MCD) spectra are reported for tetraphenyltetraacenaphthoporphyrin (TPTANP). The impact on the electronic structure of steric interactions between the fused acenaphthalene rings and the meso-tetraphenyl substituents is explored based on an analysis of the optical spectra of the Zn(II) complex (ZnTPTANP) and the free base dication species ([H4TPTANP]2+). In the case of ZnTPTANP, significant folding of the porphyrinoid ligand induces a highly unusual MCD-sign reversal providing the first direct spectroscopic evidence of ligand nonplanarity. Density functional theory (DFT) geometry optimizations for a wide range of Zn(II) porphyrinoids based on the B3LYP functional and TD-DFT calculations of the associated UV-visible absorption spectra are reported, allowing a complete assessment of the MCD data. TPTANP complexes are found to fall into a class of cyclic polyenes, termed as soft MCD chromophores by Michl (J. Pure Appl. Chem. 1980, 52, 1549.), since the signs of the Faraday A1 terms observed in the MCD spectrum are highly sensitive to slight structural changes. The origin of an unusually large red shift of the main B (or Soret) band of MTPTANP (the most red shifted ever reported for fused-ring expanded metal porphines) and of similar red shifts observed in the spectra of other peripherally crowded porphyrinoid complexes is also explored and explained on this basis. PMID- 16351100 TI - The high-valent iron-oxo species of polyoxometalate, if it can be made, will be a highly potent catalyst for C-H hydroxylation and double-bond epoxidation. AB - This study uses density functional theory (DFT) calculations to explore the reactivity of the putative high-valent iron-oxo reagent of the iron-substituted polyoxometalate (POM-FeO4-), derived from the Keggin species, PW12O40(3-). It is shown that POM-FeO4- is in principle capable of C-H hydroxylation and C=C epoxidation and that it should be a powerful oxidant, even more so than the Compound I species of cytochrome P450. The calculations indicate that in a solvent, the barriers, and especially those for epoxidation, become sufficiently small that one may expect an extremely fast reaction. An experimental investigation (by R.N. and A.M.K.) shows, however, that the formation of POM-FeO4 using the oxygen donor, F5PhI-O, leads to a persistent adduct, POM-FeO-I-PhF5(4 ), which does not decompose to POM-FeO4- + F5Ph-I at the working temperature and exhibits sluggish reactivity, in accord with previous experimental results (Hill, C. L.; Brown, R. B., Jr. J. Am. Chem. Soc. 1986, 108, 536 and Mansuy, D.; Bartoli, J.-F.; Battioni, P.; Lyon, D. K.; Finke, R. G. J. Am. Chem. Soc. 1991, 113, 7222). Subsequent calculations indeed reveal that the gas-phase binding energy of F5PhI to POM-FeO4- is high (ca. 20 kcal/mol) compared to the corresponding binding energy of propene (ca. 2-3 kcal/mol). As such, the POM-FeO I-PhF5(4-) complex is expected to be persistent toward the displacement of F5PhI by a substrate like propene, leading thereby to sluggish oxidative reactivity. According to theory, overcoming this technical difficulty may turn out to be very rewarding. The question is, can POM-FeO4- be made? PMID- 16351098 TI - Spectroscopic characterization of interstrand carbinolamine cross-links formed in the 5'-CpG-3' sequence by the acrolein-derived gamma-OH-1,N2-propano-2' deoxyguanosine DNA adduct. AB - The interstrand N2,N2-dG DNA cross-linking chemistry of the acrolein-derived gamma-OH-1,N2-propanodeoxyguanosine (gamma-OH-PdG) adduct in the 5'-CpG-3' sequence was monitored within a dodecamer duplex by NMR spectroscopy, in situ, using a series of site-specific 13C- and 15N-edited experiments. At equilibrium 40% of the DNA was cross-linked, with the carbinolamine form of the cross-link predominating. The cross-link existed in equilibrium with the non-crosslinked N2 (3-oxo-propyl)-dG aldehyde and its geminal diol hydrate. The ratio of aldehyde/diol increased at higher temperatures. The 1,N2-dG cyclic adduct was not detected. Molecular modeling suggested that the carbinolamine linkage should be capable of maintaining Watson-Crick hydrogen bonding at both of the tandem C x G base pairs. In contrast, dehydration of the carbinolamine cross-link to an imine (Schiff base) cross-link, or cyclization of the latter to form a pyrimidopurinone cross-link, was predicted to require disruption of Watson-Crick hydrogen bonding at one or both of the tandem cross-linked C x G base pairs. When the gamma-OH-PdG adduct contained within the 5'-CpG-3' sequence was instead annealed into duplex DNA opposite T, a mixture of the 1,N2-dG cyclic adduct, the aldehyde, and the diol, but no cross-link, was observed. With this mismatched duplex, reaction with the tetrapeptide KWKK formed DNA-peptide cross-links efficiently. When annealed opposite dA, gamma-OH-PdG remained as the 1,N2-dG cyclic adduct although transient epimerization was detected by trapping with the peptide KWKK. The results provide a rationale for the stability of interstrand cross-links formed by acrolein and perhaps other alpha,beta-unsaturated aldehydes. These sequence specific carbinolamine cross-links are anticipated to interfere with DNA replication and contribute to acrolein-mediated genotoxicity. PMID- 16351101 TI - Coupe du roi bisection of proteins. Spontaneous tetramerization of two peptides that span the sequence of the rabbit uteroglobin monomer. AB - The study of dividing objects into isometric segments has yielded novel approaches to the synthesis of high-symmetry organic compounds. Reported herein is the first application of this concept to a protein, rabbit uteroglobin (UG). Bisection of UG into two identical homochiral segments led to the design of the heterodimeric 70mer peptide alpha(1,2)-S-S-alpha(3,4) that spans the sequence of the native UG monomer. The ability of this compound to form a globular 140mer tetramer consisting of two noncovalently bound heterodimers was assessed by ultracentrifugation at sedimentation equilibrium and by fluorescent spectroscopy. On the other hand, the monomeric peptides alpha(1,2)-SH and alpha(3,4)-SH were shown to selectively form the alpha(1,2)-S-S-alpha(3,4) heterodimer via spontaneous air oxidation in phosphate buffer at neutral pH. PMID- 16351102 TI - Competition between glutathione and guanine for a ruthenium(II) arene anticancer complex: detection of a sulfenato intermediate. AB - The organometallic anticancer complex [(eta6-bip)Ru(en)Cl]+ (1; bip = biphenyl, en = ethylenediamine) selectively binds to guanine (N7) bases of DNA (Novakova, O.; Chen, H.; Vrana, O.; Rodger, A.; Sadler, P. J.; Brabec, V. Biochemistry 2003, 42, 11544-11554). In this work, competition between the tripeptide glutathione (gamma-L-Glu-L-Cys-Gly; GSH) and guanine (as guanosine 3',5'-cyclic monophosphate, cGMP) for complex 1 was investigated using HPLC, LC-MS and 1H,15N NMR spectroscopy. In unbuffered solution (pH ca. 3), the reaction of 1 with GSH gave rise to three intermediates: an S-bound thiolato adduct [(eta6-bip)Ru(en)(GS S)] (4) and two carboxylate-bound glutathione products [(eta6-bip)Ru(en)(GSH-O)]+ (5, 6) during the early stages (<6 h), followed by en displacement and formation of a tri-GS-bridged dinuclear Ru(II) complex [((eta6-bip)Ru)2(GS-mu-S)3]2- (7). Under physiologically relevant conditions (micromolar Ru concentrations, pH 7, 22 mM NaCl, 310 K), the thiolato complex 4 was unexpectedly readily oxidized by dioxygen to the sulfenato complex [(eta6-bip)Ru(en)(GS(O)-S)] (8) instead of forming the dinuclear complex 7. Under these conditions, competitive reaction of complex 1 with GSH and cGMP gave rise to the cGMP adduct [(eta6-bip)Ru(en)(cGMP N7)]+ (10) as the major product, accounting for ca. 62% of total Ru after 72 h, even in the presence of a 250-fold molar excess of GSH. The oxidation of coordinated glutathione in the thiolato complex 4 to the sulfenate in 8 appears to provide a facile route for displacement of S-bound glutathione by G N7. Redox reactions of cysteinyl adducts of these Ru(II) arene anticancer complexes could therefore play a significant role in their biological activity. PMID- 16351103 TI - Spin density and coenzyme M coordination geometry of the ox1 form of methyl coenzyme M reductase: a pulse EPR study. AB - Methyl-coenzyme M reductase (MCR) catalyses the reduction of methyl-coenzyme M (CH3-S-CoM) with coenzyme B (H-S-CoB) to CH4 and CoM-S-S-CoB in methanogenic archaea. Here we present a pulse EPR study of the "ready" form MCR(ox1), providing a detailed description of the spin density and the coordination of coenzyme M (CoM) to the Ni cofactor F430. To achieve this, MCR was purified from cells grown in a 61Ni enriched medium and samples were prepared in D2O with the substrate analogue CoM either deuterated in the beta-position or with 33S in the thiol group. To obtain the magnetic parameters ENDOR and HYSCORE measurements were done at X- and Q-band, and CW EPR, at X- and W-band. The hyperfine couplings of the beta-protons of CoM indicate that the nickel to beta-proton distances in MCR(ox1) are very similar to those in Ni(II)-MCR(ox1-silent), and thus the position of CoM relative to F430 is very similar in both species. Our thiolate sulfur and nickel EPR data prove a Ni-S coordination, with an unpaired spin density on the sulfur of 7 +/- 3%. These results highlight the redox-active or noninnocent nature of the sulfur ligand on the oxidation state. Assuming that MCR(ox1) is oxidized relative to the Ni(II) species, the complex is formally best described as a Ni(III) (d7) thiolate in resonance with a thiyl radical/high-spin Ni(II) complex, Ni(III)-(-)SR <--> Ni(II)-*SR. PMID- 16351104 TI - Silicon-initiated carbonylative carbotricyclization and [2+2+2+1] cycloaddition of enediynes catalyzed by rhodium complexes. AB - The reaction of dodec-11-ene-1,6-diynes or their heteroatom congeners with a hydrosilane catalyzed by Rh(acac)(CO)2 at ambient temperature and pressure of CO gives the corresponding fused 5-7-5 tricyclic products, 5-oxo-1,3a,4,5,7,9 hexahydro-3H-cyclopenta[e]azulenes or their heteroatom congeners, in excellent yields through a unique silicon-initiated cascade carbonylative carbotricyclization (CO-SiCaT) process. It has also been found that the 5-7-5 fused tricyclic products can be obtained from the same type of enediynes and CO through a novel intramolecular [2+2+2+1] cycloaddition process. The characteristics of these two tricyclization processes and the fundamental differences in their reaction mechanisms are discussed. This novel higher-order cycloaddition reaction has also been successfully applied to the tricyclization of undeca-5,10-diyn-1-als, affording the corresponding 5-7-5 fused-ring products bearing a seven-membered lactone moiety. Related [2+2+2] tricyclizations of enediyne and diynal substrates are also discussed. These newly discovered reactions can construct multiple bonds all at once, converting linear starting materials to polycyclic compounds in a single step. Thus, these new processes provide innovative routes to functionalized polycyclic compounds that are useful for the syntheses of natural and unnatural products. PMID- 16351105 TI - Enthalpic pair interaction coefficient between zwitterions of L-alpha-amino acids and urea molecule as a hydrophobicity parameter of amino acid side chains. AB - Dissolution enthalpies of L-alpha-proline, L-alpha-tyrosine, L-alpha-tryptophan, L-alpha-histidyne, L-alpha-arginine, L-alpha-lysine, L-aspartic acid, and L-alpha glutamic acid in aqueous solutions of urea have been measured by calorimetry at a temperature of 298.15 K. The values of dissolution enthalpy were used to determine enthalpic heterogeneous pair interaction coefficients between the zwitterions of the natural amino acids and a molecule of urea in water solution. These coefficients were interpreted in terms of the hydrophobic or hydrophilic effects of the side chains of amino acids on their interactions with a polar molecule of urea in water. PMID- 16351106 TI - Heat of hydrogenation of 1,5-dehydroquadricyclane. A computational and experimental study of a highly pyramidalized alkene. AB - The radical anion of the highly pyramidalized alkene 1,5-dehydroquadricyclane (1) was generated in the gas phase from the Squires reaction of 1,5 bis(trimethylsilyl)quadricyclane with F-/F2. The electron binding energy and proton affinity of 1*- were determined by bracketing experiments to be 0.6 +/- 0.1 eV and 386 +/- 5 kcal/mol, respectively. These values are in good agreement with values predicted by density functional theory (B3LYP/6-31+G*) and ab initio (CASPT2/6-31+G*) calculations. The experimental heat of hydrogenation of 1, obtained from a thermochemical cycle, was found to be 91 +/- 9 kcal/mol. This value of deltaH(H2) leads to values of 67 +/- 9 kcal/mol for the olefin strain energy (OSE) of 1, 172 +/- 9 kcal/mol for its heat of formation, and 23 +/- 9 kcal/mol for its pi bond dissociation enthalpy. Since the retro-Diels-Alder reaction of neutral 1 is computed to be highly exothermic, the finding that 1*- apparently does not undergo a retro-Diels-Alder reaction is of particular interest. The B3LYP/6-31+G* optimized geometry of 1 suggests that the bonding in this alkene is partially delocalized, presumably because the highly pyramidalized double bond in 1 interacts with the distal cyclopropane bonds in a manner that eventually leads to a retro-Diels-Alder reaction. The good agreement of the B3LYP and (2/2)CASPT2 values for the heat of hydrogenation and OSE of 1 with the experimentally derived values provides indirect evidence for the correctness of the B3LYP prediction that the equilibrium geometry of 1 lies part way along the reaction coordinate to the transition structure for the retro-Diels-Alder reaction. PMID- 16351108 TI - Generalized anomeric interpretation of the "high-energy" N-P bond in N-methyl-N' phosphorylguanidine: importance of reinforcing stereoelectronic effects in "high energy" phosphoester bonds. AB - Electronic structure calculations have been performed on a model N phosphorylguanidine, or phosphagen, to understand the stereoelectronic factors contributing to the lability of the "high-energy" N-P bond. The lability of the N P bond is central to the physiological role of phosphagens involving phosphoryl transfer reactions important in cellular energy buffering and metabolism. Eight protonated forms of N-methyl-N'-phosphorylguanidine have been energy minimized at levels of theory ranging up to B3LYP/6-311++G(d,p) and MP2/6-311++G(d,p) to investigate the correlation between protonation state and N-P bond length. Selected forms have also been minimized using the CCSD/6-311++G(d,p) and QCISD/6 311++G(d,p) levels of theory. Bulk solvation energies using the polarized continuum model (PCM) with B3LYP/6-311++G(d,p) test the influence of the surroundings on computed structures and energies. The N-P bond length depends on the overall protonation state where increased protonation at the phosphoryl group or deprotonation at the unsubstituted N'' nitrogen results in shorter, stronger N P bonds. Natural bond orbital analysis shows that the protonation state affects the N-P bond length by altering the magnitude of stabilizing n(O) --> sigma*(N-P) stereoelectronic interactions and to a lesser extent the sigma(N-P) --> sigma*(C N'') and sigma(N-P) --> sigma*(C-N) interactions. The computations do not provide evidence of a competition between the phosphoryl and guanidinium groups for the same lone pair on the bridging nitrogen, as previously suggested by opposing resonance theory. The computed n(O) --> sigma*(N-P) anomeric effect provides a novel explanation of "high-energy" N-P bond lability. This offers new mechanistic insight into phosphoryl transfer reactions involving both phosphagens and other biochemically important "high-energy" phosphoester bonds. PMID- 16351107 TI - Oxidative cyclizations in a nonpolar solvent using molecular oxygen and studies on the stereochemistry of oxypalladation. AB - Oxidative cyclizations of a variety of heteroatom nucleophiles onto unactivated olefins are catalyzed by palladium(II) and pyridine in the presence of molecular oxygen as the sole stoichiometric oxidant in a nonpolar solvent (toluene). Reactivity studies of a number of N-ligated palladium complexes show that chelating ligands slow the reaction. Nearly identical conditions are applicable to five different types of nucleophiles: phenols, primary alcohols, carboxylic acids, a vinylogous acid, and amides. Electron-rich phenols are excellent substrates, and multiple olefin substitution patterns are tolerated. Primary alcohols undergo oxidative cyclization without significant oxidation to the aldehyde, a fact that illustrates the range of reactivity available from various Pd(II) salts under differing conditions. Alcohols can form both fused and spirocyclic ring systems, depending on the position of the olefin relative to the tethered alcohol; the same is true of the acid derivatives. The racemic conditions served as a platform for the development of an enantioselective reaction. Experiments with stereospecifically deuterated primary alcohol substrates rule out a "Wacker-type" mechanism involving anti oxypalladation and suggest that the reaction proceeds by syn oxypalladation for both mono- and bidentate ligands. In contrast, cyclizations of deuterium-labeled carboxylic acid substrates undergo anti oxypalladation. PMID- 16351109 TI - Design and synthesis of intramolecular charge transfer-based fluorescent reagents for the highly-sensitive detection of proteins. AB - Novel fluorescent molecular probes possessing both a hydroxystyryl and a cyanopyranyl moieties were designed and synthesized to detect the proteins via noncovalent bonding. These fluorescent probes indicated very weak fluorescence emission in the absence of protein. On the other hand, the fluorescence spectra of these probes showed a large Stokes shift and dramatic increase of fluorescence intensity, and red emission was observed after addition of BSA. These fluorescence spectral changes upon binding proteins were caused by the ICT process. Fluorescence intensities of the probes were plotted as a function of protein concentrations. A good linear relationship was observed up to 1000 microg/mL of protein, and the detection limit was found to be 100 ng/mL at the given assay conditions. Similar results were observed for the measurements of not only BSA but also other proteins (BGG, etc.). The responses of these probes to various nonprotein substances (inorganic salts, chelating agents, etc.) were observed, the fluorescence intensity did not change before and after the addition of foreign substances, and correct protein monitoring was successful using these fluorescent probes. To demonstrate the application of these probes, proteins after the separation using SDS-PAGE were stained in the medium containing 1, and the imaging of the proteins in the gel was successful. The experimental results clearly showed that these probes are good protein indicators for easy and highly sensitive detection. PMID- 16351110 TI - Nano- and microscopic surface wrinkles of linearly increasing heights prepared by periodic precipitation. AB - Arrays of surface wrinkles of linearly increasing heights (from tens of nanometers to tens of micrometers) were prepared via a spontaneous reaction diffusion process based on periodic precipitation. The slopes, dimensions, and positions of the precipitation bands could be controlled precisely by adjusting the concentrations of the participating chemicals as well as the material properties of patterned substrates. Additional control of periodic precipitation by localized UV irradiation allowed for the preparation of discontinuous and curvilinear structures. The nonbinary 3D surface topographies were replicated into poly(dimethylsiloxane), and the applications of replicas in microfluidics, microseparations, and cell biology have been suggested. PMID- 16351111 TI - Unique layered double hydroxide morphologies using reverse microemulsion synthesis. AB - We report the first controlled synthesis of a layered double hydroxide (LDH) in a water-in-oil reverse microemulsion system. This synthesis of Mg2Al-LDHs was carried out in the reverse microemulsion of NaDDS (sodium dodecyl sulfate)-water isooctane with water/surfactant molar ratio = 24. This enables us to obtain nanometer sized LDH platelets typically with a 40-50 nm diameter and 10 nm thickness. Further modification of the reverse microemulsion using triblock copolymers during crystallization allowed us to express different growth orientations of the LDH structure. These data show that the aspect ratio of LDHs can be flexibly adjusted over a wide range. PMID- 16351113 TI - Properties of a mixed-valence (Fe(II))2(Fe(III))2 square cell for utilization in the quantum cellular automata paradigm for molecular electronics. AB - The di-mixed-valence complex [{(eta(5)-C5H5)Fe(eta(5)-C5H4)}4(eta(4)-C4)Co(eta(5) C5H5)]2+, 1(2+), has been evaluated as a molecular four-dot cell for the quantum cellular automata paradigm for electronic devices. The cations 1(1+) and 1(2+) are prepared in good yield by selective chemical oxidation of 1(0) and are isolated as pure crystalline materials. The solid-state structures of 1(0) and 1(1+) and the midrange- and near-IR spectra of 1(0), 1(1+), 1(2+), and 1(3+) have been determined. Further, the variable-temperature EPR spectra of 1(1+) and 1(2+), magnetic susceptibility of 1(1+) and 1(2+), Mossbauer spectra of 1(0), 1(1+), and 1(2+), NMR spectra of 1(0), and paramagnetic NMR spectra of 1(1+) and 1(2+) have been measured. The X-ray structure determination reveals four ferrocene "dots" arranged in a square by C-C bonds to the corners of a cyclobutadiene linker. The four ferrocene units project from alternating sides of the cyclobutadiene ring and are twisted to minimize steric interactions both with the Co(eta(5)-C5H5) fragment and with each other. In the solid state 1(2+) is a valence-trapped Robin and Day class II compound on the 10(-12) s infrared time scale, the fastest technique used herein, and unambiguous evidence for two Fe(II) and two Fe(III) sites is observed in both the infrared and Mossbauer spectra. Both EPR and magnetic susceptibility measurements show no measurable spin-spin interaction in the solid state. In solution, the NMR spectra show that free rotation around the C-C bonds connecting the ferrocene units to the cyclobutadiene ring becomes increasingly hindered with decreasing temperature, leading to spectra at the lowest temperature that are consistent with the solid state structure. Localization of the charges in the cations, which is observed in the paramagnetic NMR spectra as a function of temperature, correlates with the fluxional behavior. Hence, the alignment between the pi systems of the central linker and the ferrocene moieties most likely controls the rate of electron exchange between the dots. PMID- 16351112 TI - RNA-mediated control of metal nanoparticle shape. AB - RNA sequences previously isolated by in vitro selection were further characterized for their ability to control palladium particle growth. Five pyridyl-modified RNA sequences (Pdases) representing each of the different evolved families were found to form hexagonal plates with a high degree of shape specificity. However, a sixth nonrelated pyridyl-modified RNA sequence was found to form exclusively cubic particles under identical conditions. Replacing pyridyl modified RNA with native RNA resulted in a complete loss of RNA function. Removing the 3'-fixed sequence region from the Pdase had little effect on particle growth; however, further truncations into the variable region resulted in a significant loss of activity and particle shape control. These Pdases were selected using the organometallic precursor complex tris(dibenzylideneacetone) dipalladium(0) ([Pd2(DBA)3]). Changing the metal center and ligand of the group VIII organometallic precursor complex revealed a strong dependence of particle growth and shape on the DBA ligands. Changing the metal center from Pd to Pt while retaining the DBA ligands gave predominantly hexagonal Pt, but with a decrease in shape control. Taken together, the results of this study suggest that the full-length Pdases contain active sites capable of highly specific molecular recognition of organometallic complexes as particle formation reagents. PMID- 16351114 TI - Organic syntheses on an icosahedral borane surface: closomer structures with twelvefold functionality. AB - The syntheses of a series of novel ester-linked derivatives of the icosahedral [closo-B12(OH)12]2- boron cluster (closomer esters) are described using several synthetic methods. The reaction of bis(tetrabutylammonium)-closo-dodecahydroxy dodecaborate, [NBu4]2 1, with carboxylic acid chlorides and anhydrides, vinyl esters with a Y5(OiPr)13O catalyst and 1,1'-carbonyldiimidazole-activated carboxylic acids yields the corresponding dianionic dodeca-ester closomers. The method using 1,1'-carbonyldiimidazole-activated carboxylic acids may be employed as a general synthetic strategy. The use of elevated reaction temperatures, achievable under pressure, to expedite syntheses is described. An attractive methodology using immobilized scavenger reagents for the expeditious purification of the closomer esters was employed. The developed methodology is compatible with a variety of peripheral functional groups attached to the termini of densely packed, carboxylate ester-linked radial arms bonded to the icosahedral borane surface. A closomer ester having twelve terminal amino groups was prepared, and without isolation, fully acetylated in good yield. PMID- 16351115 TI - Why is the partial molar volume of CO2 so small when dissolved in a room temperature ionic liquid? Structure and dynamics of CO2 dissolved in [Bmim+] [PF6(-)]. AB - When supercritical CO2 is dissolved in an ionic liquid, its partial molar volume is much smaller than that observed in most other solvents. In this article we explore in atomistic detail and explain in an intuitive way the peculiar volumetric behavior experimentally observed when supercritical CO2 is dissolved in 1-butyl-3-methylimidazolium hexafluorophosphate ([Bmim+] [PF6(-)]). We also provide physical insight into the structure and dynamics occurring across the boundary of the CO2 ionic liquid interface. We find that the liquid structure of [Bmim+] [PF6(-)] in the presence of CO2 is nearly identical to that in the neat ionic liquid (IL) even at fairly large mole fractions of CO2. Our simulations indicate, in agreement with experiments, that partial miscibilities of one fluid into the other are very unsymmetrical, CO2 being highly soluble in the ionic liquid phase while the ionic liquid is highly insoluble in the CO2 phase. We interpret our results in terms of the size and shape of spontaneously forming cavities in the ionic liquid phase, and we propose that CO2 occupies extremely well-defined locations in the IL. Even though our accurate prediction of cavity sizes in the neat IL indicates that these cavities are small compared with the van der Waals radius of a single carbon or oxygen atom, CO2 appears to occupy a space that was for the most part a priori "empty". PMID- 16351116 TI - Properties of inhibitors of methane hydrate formation via molecular dynamics simulations. AB - Within the framework of a proposed two-step mechanism for hydrate inhibition, the energy of binding of four inhibitor molecules (PEO, PVP, PVCap, and VIMA) to a hydrate surface is estimated with molecular dynamic simulations. One key feature of this proposed mechanism is that the binding of an inhibitor molecule to the surface of an ensuing hydrate crystal disrupts growth and therein crystallization. It is found through the molecular dynamic simulations that inhibitor molecules that experimentally exhibit better inhibition strength also have higher free energies of binding, an indirect confirmation of our proposed mechanism. Inhibitors increasing in effectiveness, PEO < PVP < PVCap < VIMA, have increasingly negative (exothermic) binding energies of -0.2 < -20.6 < -37.5 < 45.8 kcal/mol and binding free energies of increasing favorability (+0.4 approximately = +0.5 < -9.4 < -15.1 kcal/mol). Furthermore, the effect of an inhibitor molecule on the local liquid water structure under hydrate-forming conditions was examined and correlated to the experimental effectiveness of the inhibitors. Two molecular characteristics that lead to strongly binding inhibitors were found: (1) a charge distribution on the edge of the inhibitor that mimics the charge separation in the water molecules on the surface of the hydrate and (2) the congruence of the size of the inhibitor with respect to the available space at the hydrate-surface binding site. Equipped with this molecular level understanding of the process of hydrate inhibition via low-dosage kinetic hydrate inhibitors we can design new, more effective inhibitor molecules. PMID- 16351117 TI - Measuring site-specific cluster-surface bond formation. AB - Recent advances in dynamic force microscopy show that it is possible to measure the forces between atomically sharp tips and particular atomic positions on surfaces as a function of distance. However, on most ionic surfaces, the positive and negative ions can so far not be distinguished. In this paper, we use the CaF2(111) surface, where atomic resolution force microscopy has allowed identification of the positions of the Ca2+ and F- ions in the obtained images, to demonstrate that short-range interaction forces can be measured selectively above chemically identified surface sites. Combining experimental and theoretical results allows a quantification of the strength and distance dependence of the interaction of a tip-terminating cluster with particular surface ions and reveals details of cluster and surface relaxation. Further development of this approach will provide new insight into mechanisms of chemical bond formation between clusters, cluster deposition at surfaces, processes in adhesion and tribology, and single atom manipulation with the force microscope. PMID- 16351118 TI - Solvent friction effect on intramolecular electron transfer. AB - U-shaped donor-bridge-acceptor molecules with different electronic couplings have been investigated as a function of temperature in solvents with slow polarization relaxation, in particular, N-methylacetamide (NMA) and N-methylpropionamide (NMP). At high temperature, the electron-transfer rate is well described by a nonadiabatic model; however, the rate at low temperature is controlled by the solvent friction. The change of the electron-transfer mechanism is discussed and compared with theoretical models. PMID- 16351119 TI - Branched coordination multilayers on gold. AB - A C3-symmetric tridentate hexahydroxamate ligand molecule was specially synthesized and used for coordination self-assembly of branched multilayers on Au surfaces precoated with a self-assembled monolayer (SAM) of ligand anchors. Layer by-layer (LbL) growth of multilayers via metal-organic coordination using Zr4+ ions proceeds with high regularity, adding one molecular layer in each step, as shown by ellipsometry, wettability, UV-vis spectroscopy, and atomic force microscopy (AFM). The branched multilayer films display improved stiffness, as well as a unique defect self-repair capability, attributed to cross-linking in the layers and lateral expansion over defects during multilayer growth. Transmetalation, i.e., exposure of Zr4+-based assemblies to Hf4+ ions, was used to evaluate the cross-linking. Conductive atomic force microscopy (AFM) was used to probe the electrical properties of the multilayers, revealing excellent dielectric behavior. The special properties of the branched layers were emphasized by comparison with analogous multilayers prepared similarly using linear (tetrahydroxamate) ligand molecules. The process of defect annihilation by bridging over defective areas, attributed to lateral expansion via the excess bishydroxamate groups, was demonstrated by introduction of artificial defects in the anchor monolayer, followed by assembly of two layers of either the linear or the branched molecule. Analysis of selective binding of Au nanoparticles (NPs) to unblocked defects emphasized the superior repair mechanism in the branched layers with respect to the linear ones. PMID- 16351120 TI - Bronsted base-modulated regioselectivity in the aerobic oxidative amination of styrene catalyzed by palladium. AB - Palladium(II)-catalyzed aerobic oxidative amination of styrene with oxazolidinone proceeds with catalyst-controlled regioselectivity: (CH3CN)2PdCl2 (1) and (Et3N)2PdCl2 (2) catalyze formation of the anti-Markovnikov and Markovnikov enecarbamate products, 3 and 4, respectively. Kinetic studies and deuterium kinetic isotope effects demonstrate that these two reactions possess different rate-limiting steps, and the data indicate that the product regiochemistry arises from the presence or absence of an effective Bronsted base in the reaction. In the presence of a Bronsted base such as triethylamine or acetate, the kinetically preferred Markovnikov aminopalladation adduct of styrene is trapped via rapid deprotonation of a zwitterionic intermediate and leads to formation of 4. In the absence of an effective Bronsted base, however, slow deprotonation of this adduct enables aminopalladation to be reversible, and product formation proceeds through the thermodynamically preferred anti-Markovnikov aminopalladation adduct to yield 3. PMID- 16351121 TI - Oligomeric cholates: amphiphilic foldamers with nanometer-sized hydrophilic cavities. AB - The hydroxyl at the C-3 of cholic acid was converted to an amino group, and the resulting amino-functionalized cholic acid was used as a monomer to prepare amide linked oligomeric cholates. These cholate oligomers fold into helical structures with nanometer-sized hydrophilic internal cavities in solvent mixtures consisting of mostly nonpolar solvents such as carbon tetrachloride or ethyl acetate/hexane and 2-5% of a polar solvent such as methanol or DMSO. The conformations of the foldamers were studied by UV, fluorescence, fluorescence quenching, and fluorescence resonance energy transfer. The nature of the polar/nonpolar solvents and their miscibility strongly influenced the folding reaction. Folding was cooperative, as evidenced by the sigmoidal curves in solvent denaturation experiments. The folded conformers became more stable with an increase in the chain length. The folding/unfolding equilibrium was highly sensitive toward the amount of polar solvent. One percent variation in the solvent composition could change the folding free energies by 0.5-1.4 kcal/mol. PMID- 16351122 TI - Programming the internal structure and stability of helical pores self-assembled from dendritic dipeptides via the protective groups of the peptide. AB - The synthesis of dendritic dipeptides (4-3,4-3,5)12G2-CH2-X-L-Tyr-L-Ala-OMe with X = Boc, Moc, and Ac; their self-assembly in bulk and in solution; and the structural and retrostructural analysis of their supramolecular helical porous assemblies are reported. The dimensions, structure, internal order, thermal stability of the supramolecular helical pores, and conformations of the dendron and supramolecular dendrimer are programmed by the nature of the protective groups of the dipeptide. The ability of the protective groups to program the structure of the helical pore reveals the simplest design strategy that complements the more complex strategies based on the architecture of the dendron, the stereochemistry, and the structure of the dipeptide. PMID- 16351123 TI - Fully stereocontrolled total syntheses of the prostacyclin analogues 16S-iloprost and 16S-3-oxa-iloprost by a common route, using alkenylcopper-azoalkene conjugate addition, asymmetric olefination, and allylic alkylation. AB - In this article we describe fully stereocontrolled total syntheses of 16S iloprost (16S-2), the most active component of the drugs Ilomedin and Ventavis, and of 16S-3-oxa-iloprost (16S-3), a close analogue of 16S-2 having the potential for a high oral activity, by a new and common route. The key steps of this route are (1) the establishment of the complete C13-C20 omega side chain of the target molecules through a stereoselective conjugate addition of the alkenylcopper derivative 9 to the bicyclic C6-C12 azoalkene 10 with formation of hydrazone 8, (2) the diastereoselective olefination of ketone 7 with the chiral phosphoryl acetate 39, and (3) the regio- and stereoselective alkylation of the allylic acetate 43 with cuprate 42. These measures allowed the 5E,15S,16S-stereoselective synthesis of 16S-2 and 16S-3, a goal which had previously not been achieved. Azoalkene 10 was obtained from the achiral bicyclic C6-C12 ketone 11 as previously described by using as key step an enantioselective deprotonation. The configuration at C16 of omega-side chain building block 9 has been installed with high stereoselectivity by the oxazolidinone method and that at C15 by a diastereoselective oxazaborolidine-catalyzed reduction of the C13-C20 ketone 23 with catecholborane. Surprisingly, a high diastereoselectivity in the reduction of 23 was only obtained by using 2 equiv of oxazaborolidine 24. Application of substoichiometric amounts of 24 resulted in irreproducible diastereoselectivities ranging from very high to nil. PMID- 16351125 TI - Fesulphos-palladium(II) complexes as well-defined catalysts for enantioselective ring opening of meso heterobicyclic alkenes with organozinc reagents. AB - The air-stable and readily available cationic methyl palladium(II) complexes of planar chiral Fesulphos ligands [(Fesulphos)Pd(Me)(PhCN)]+ X- are highly efficient catalysts for the alkylative ring opening of oxa- and azabicyclic alkenes with dialkylzinc reagents, showing broad scope with regards to both the bicyclic substrate and the dialkylzinc reagent. Catalyst loading as low as 0.5 mol % is sufficient to achieve good yields and enantioselectivities ranging 94 - > 99% ee in most cases. {Fesulphos = (1-phosphino-2-sulfenylferrocene); X- = tetrakis[3,5-bis(trifluoromethyl)phenyl]borate or PF6(-)].} The origin of the high asymmetric induction has been rationalized by mechanistic studies combining computational calculations and X-ray structural analysis. PMID- 16351124 TI - Ru-catalyzed alkene-alkyne coupling. Total synthesis of amphidinolide P. AB - A coordinatively unsaturated ruthenium complex catalyzed the formation of a carbon-carbon bond between two judiciously chosen alkene and alkyne partners in good yield, and in a chemo- and regioselective fashion, despite the significant degree of unsaturation of the substrates. The resulting 1,4-diene forms the backbone of the cytotoxic marine natural product amphidinolide P. The alkene partner was rapidly assembled from (R)-glycidyl tosylate, which served as a linchpin in a one-flask, sequential three-components coupling process using vinyllithium and a vinyl cyanocuprate. The synthesis of the alkyne partner made use of an unusual anti-selective addition under chelation-control conditions of an allyltin reagent derived from tiglic acid. In addition, a remarkably E selective E2 process using the azodicarboxylate-triphenylphosphine system is featured. Also featured is the first example of the use of a beta-lactone as a thermodynamic spring to effect macrolactonization. The oxetanone ring was thus used as a productive protecting group that increased the overall efficiency of this total synthesis. This work was also an opportunity to further probe the scope of the ruthenium-catalyzed alkene-alkyne coupling, in particular using enynes, and studies using various functionalized substrates are described. PMID- 16351126 TI - Theoretical study of the 13C NMR spectroscopy of single-walled carbon nanotubes. AB - The 13C NMR spectroscopy of armchair and zigzag single-walled carbon nanotubes has been investigated theoretically. Spectra for (4,4), (5,5), (6,6), (6,0), (9,0), and (10,0) nanotubes have been simulated based on ab initio calculations of model systems. The calculations predict a dominant band arising from the carbon atoms in the "tube" with smaller peaks at higher chemical shifts arising from the carbon atoms of the caps. The dominant band lies in the range of 128 and 138 ppm. Its position depends weakly on the length, width, and chirality of the tubes. The calculations demonstrate how structural information may be gleaned from relatively low-resolution nanotube 13C NMR spectra. PMID- 16351127 TI - Oxygen- and temperature-dependent kinetic isotope effects in choline oxidase: correlating reversible hydride transfer with environmentally enhanced tunneling. AB - Choline oxidase catalyzes the flavin-linked oxidation of choline to glycine betaine, with betaine aldehyde as intermediate and oxygen as electron acceptor. Here, the effects of oxygen concentration and temperature on the kinetic isotope effects with deuterated choline have been investigated. The D(kcat/Km) and Dkcat values with 1,2-[(2)H4]-choline were pH-independent at saturating oxygen concentrations, whereas they decreased at high pH to limiting values that depended on oxygen concentration at < or = 0.97 mM oxygen. The kcat/Km and kcat pH profiles had similar patterns reaching plateaus at high pH. Both the limiting kcat/Km at high pH and the pKa values were perturbed to lower values with choline and < or = 0.25 mM oxygen. These data suggest that oxygen availability modulates whether the reduced enzyme-betaine aldehyde complex partitions forward to catalysis rather then reverting to the oxidized enzyme-choline alkoxide species. At saturating oxygen concentrations, the D(kcat/Km) was 10.6 +/- 0.6 and temperature independent, and the isotope effect on the preexponential factors (A(H)'/A(D)') was 14 +/- 3, ruling out a classical over-the-barrier behavior for hydride transfer. Similar enthalpies of activation (deltaH(double dagger)) with values of 18 +/- 2 and 18 +/- 5 kJ mol(-1) were determined with choline and 1,2 [(2)H4]-choline. These data suggest that the hydride transfer reaction in which choline is oxidized by choline oxidase occurs quantum mechanically within a preorganized active site, with the reactive configuration for hydride tunneling being minimally affected by environmental vibrations of the reaction coordinate other than those affecting the distance between the donor and acceptor of the hydride. PMID- 16351129 TI - Quantitative analysis of protein phosphorylation in mouse brain by hypothesis driven multistage mass spectrometry. AB - Determination of site-specific changes in the levels of protein phosphorylation in mammals presents a formidable analytical challenge. Here, we demonstrate a strategy for such analyses utilizing a combination of stable isotope chemical labeling and tandem mass spectrometry. Phosphoproteins of interest are isolated from two sets of animals that have undergone differential drug treatments, separated by SDS-PAGE, excised, and subjected to in-gel enzymatic digestion. Using a simple chemical labeling step, we introduce stable, isotopically distinct mass tags into each of the two sets of peptides that originate from the samples under comparison, mix the samples, and subject the resulting mixture to a procedure based on our previously reported hypothesis-driven multistage MS (HMS MS) method (Chang, E. J.; Archambault, V.; McLachlin, D. T.; Krutchinsky, A. N.; Chait, B. T. Anal. Chem. 2004, 76, 4472-4483). The method takes advantage of the dominant loss of H3PO4 during MS/MS from singly charged phosphopeptide ions produced by matrix-assisted laser desorption/ionization (MALDI) in the ion trap mass spectrometer. In the present work, quantitation is achieved by isolating the range of m/z values that include both isotopic forms of the putative phosphopeptide and measuring the relative intensities of the two resulting -98-Da fragment ion peaks. This MS/MS measurement can be repeated on the same MALDI sample for all potential phosphopeptide ion pairs that we hypothesize might be produced from the protein under study. Use of MS/MS for quantitation greatly increases the sensitivity of the method and allows us to measure relatively low levels of phosphorylation, phosphopeptides, or both that are not easily observable by single-stage MS. We apply the current method to the determination of changes in the levels of phosphorylation in DARPP-32 from the mouse striatum upon treatment of animals with psychostimulant drugs. PMID- 16351130 TI - Molecular dynamics simulations of alkylsilane stationary-phase order and disorder. 1. Effects of surface coverage and bonding chemistry. AB - "Shape-selective" polymeric alkylsilane stationary phases are routinely employed over the more common monomeric phases in reversed-phase liquid chromatography (RPLC) to improve the separation of geometric isomers of shape-constrained solutes. We have investigated the molecular dynamics of chromatographic models that represent both monomeric and polymeric stationary phases with alkylsilane surface coverages and bonding chemistries typical of actual materials in an effort to elucidate the molecular-level structural features that control shape selective separations. The structural characterization of these models is consistent with previous experimental observations of alkyl chain order and disorder: (1) alkyl chain order increases with increased surface coverage; and (2) monomeric and polymeric phases with similar surface coverages yield similar alkyl chain order (although subtle differences exist). In addition, a significant portion of the alkyl chain proximal to the silica surface is disordered (primarily gauche conformations) and the distal end is most ordered. Models that represent shape-selective RPLC phases possess a significant region of distal end chain order with primarily trans dihedral angle conformations. This is consistent with the view that the alkyl chains comprising polymeric stationary phases contain a series of well-defined and rigid voids in which shape-constrained solutes can penetrate and hence be selectively retained. PMID- 16351131 TI - Molecular dynamics simulations of alkylsilane stationary-phase order and disorder. 2. Effects of temperature and chain length. AB - In an effort to elucidate the molecular-level structural features that control shape-selective separations, we have investigated the molecular dynamics of chromatographic models that represent both monomeric and polymeric stationary phases with alkylsilane length and temperature conditions analogous to actual materials of low to high shape selectivity. The structural characterization of these models is consistent with previous experimental observations of alkyl chain order and disorder: alkyl chain order increases both with alkyl chain length and with reduced temperature. Models that represent shape-selective reversed-phase liquid chromatography (RPLC) phases possess a significant region of distal end chain order with primarily trans dihedral angle conformations; the extension of these ordered regions into the phase increases with an increase in chain length. Models with extended chain length (C30) possess a higher degree of conformational order and are relatively insensitive to changes in surface coverage, bonding chemistry, and temperature. Chromatography models of various chain lengths and over a temperature range that represents highly shape-selective RPLC stationary phases all contain a series of well-defined and rigid cavities; the size and depth of these "slots" increase for the C30 models, which may promote the enhanced separations of larger size shape-constrained solutes, such as carotenoids. PMID- 16351132 TI - Dual-capillary backscatter interferometry for high-sensitivity nanoliter-volume refractive index detection with density gradient compensation. AB - A simple, stable, ultrasensitive dual-capillary dual-bicell (DCDB) microinterferometic backscattering detection (MIBD) system was developed. In DCDB MIBD, a He-Ne laser beam passes through a half-wave plate onto the cross section of two capillaries, one for reference and another for sensing analyte. The position of the backscattered fringe from each capillary, which are in proximity or essentially identical thermal environments, was detected with matched bicell photodetectors. The configuration was found to effectively compensate for thermal drift, which is normally the major source of noise in refractive index (RI) detection systems. It is shown that passive environmental compensation leads to greatly enhanced signal in nanoscale refractometry preformed by MIBD. An order of magnitude improvement in detection limits over single channel configurations is possible. Performance reaches the 10(-9) RIU level for like solvents in the presence in extremely large thermally induced RI gradients. At this level of detectability, DCDB MIBD could facilitate nanoliter-volume, femtomole-level universal detection in applications ranging from mu-HPLC and on-chip CE to scanning microcalorimetry. PMID- 16351133 TI - Multistage isoelectric focusing in a polymeric microfluidic chip. AB - This paper reports a protocol that improves the resolving power of isoelectric focusing (IEF) in a polymeric microfluidic chip. This method couples several stages of IEF in series by first focusing proteins in a straight channel using broad-range ampholytes and then refocusing segments of the first channel into secondary channels that branch from the first one at T-junctions. Experiments demonstrate that several fluorescent proteins that had focused within a segment of the straight channel in the first stage were refocused at significantly higher resolution due to the shallower pH gradient and higher electrical field gradient. Two variants of green fluorescent protein from the second-stage IEF fractionation were further separated in a third stage. Three stages of IEF were completed in less than 25 min at electric field strengths ranging from 50 to 214 V/cm. PMID- 16351134 TI - Multiphenotypic whole-cell sensor for viability screening. AB - Here, we describe the fabrication of whole mammalian cell biosensors for the optical monitoring of cell viability. Three phenotypes were examined for their response to the addition of two model chemotoxins: sodium hypochlorite and sodium azide, and one model biotoxin, concanavalin A. Two sensing platforms containing cells, hydrogel microspheres, or hydrogel arrays, were also explored. Step changes in viability in response to small doses of sodium hypochlorite were seen nearly instantaneously in all cell lines, in solution, microspheres, and microarrays. Linear detection of sodium azide by entrapped hepatocytes was 0-10 microM, whereas the linear detection range for macrophages and endothelial cells was 0-75 microM. Macrophages and hepatocytes have a greater sensitivity, as indicated by a 40% change in fluorescence over the linear range, whereas endothelial cells show only a 15% change in fluorescence over the linear range. Using photoreaction injection molding, we were also able to generate a multiphenotype sensor that enables the measurement of the toxic effect of 100 microg/mL concanavalin A on macrophages and hepatocytes, but not on endothelial cells. PMID- 16351135 TI - High-temperature liquid chromatography coupled on-line to a continuous-flow biochemical screening assay with electrospray ionization mass spectrometric detection. AB - The potential of high-temperature liquid chromatography (HTLC) was investigated in an on-line combination with a screening system for bioactive compounds against the enzyme cathepsin B. Samples were separated by HTLC and subsequently analyzed by an on-line continuous-flow enzymatic assay. Detection was performed by electrospray ionization mass spectrometry, revealing both the bioactivity and the molecular mass of the bioactive compounds. Compared to conventional reversed phase liquid chromatography, the amount of methanol necessary for separation could be decreased to only 10%, which improved the compatibility of LC with a biochemical assay. Sufficient preheating of the mobile phase prior to the separation and postcolumn cooling to prevent deactivation of the enzyme, even at column temperatures as high as 208 degrees C, was achieved as indicated by the reliable peak shapes obtained. The sensitivity was comparable with previously described systems operating at ambient temperatures as similar IC50 values were obtained. Exposing the inhibitors to high temperatures did not lead to thermal decomposition. The separation of inhibitors and the subsequent biochemical assay was performed either isothermally at various temperatures or by applying various temperature gradients as well as at various flow rates. The results obtained clearly show the compatibility of HTLC with an enzymatic screening assay. PMID- 16351136 TI - Functional membrane-implanted lab-on-a-chip for analysis of percent HDL cholesterol. AB - A functional lab-on-a-chip has been developed for simultaneous quantitative analyses of high-density lipoprotein (HDL) cholesterol (HDL-C) and total cholesterol (total-C) in a submicroliter plasma sample. The analytical device was fabricated by placing commercial membranes, traditionally used for rapid diagnostics, within microfluidic channels engraved on the surface of a plastic chip. The concentration of HDL-C was measured using enzymatic reactions to produce a colorimetric signal after separation of the single plasma lipoprotein from a mixture. Two small pieces of different membrane pads were used to provide each group of reagents, for HDL separation and enzyme reactions, deposited within their tiny pores in a dry state. To maintain a connection toward the capillary action of the medium, the pads were arranged in a sequence within the fluidic channel that controlled the inlet and outlet of the flow. Upon the addition of a sample, the fluid was delivered through the pads of the chip and a color signal was subsequently generated in proportion to the concentration of HDL-C. The level of total-C was concurrently determined by following identical processes, except absent HDL separation. The two signals were simultaneously determined by employing optical detectors based on transmittance of a light. Such total analyses were completed within 2 min, and the sample sizes were able to be reduced to 0.4 microL for HDL-C and 0.1 microL for total-C, enough to cover the clinically required dynamic ranges. PMID- 16351137 TI - Smart chemical taste sensor for determination and prediction of taste qualities based on a two-phase optimized radial basis function network. AB - A novel, smart, chemical taste sensor that realistically mimics the behavior of the human gustatory system is described. The taste sensor consists of an array of electrochemical sensors that represent the gustatory receptors on the human tongue, and a two-phase optimized radial basis function network (RBFN) to represent the human brain, which comprehensively analyzes the gustatory stimulation and judges the overall taste. In the illustrated model, eight electrodes were fabricated to determine the eight major taste-causing substances, Na+, K+, Cl-, H+, sucrose, glucose, glutamate, and caffeine. The detected signals were fed to a two-phase RBFN optimized by the implementation of a basis optimization algorithm and weight decay term for appropriate data processing. The first phase of the two-phase RBFN quantifies the amount of taste-causing substances in food samples from the responses of the electrodes. These results are then fed to the second phase, which correlates the amount of substances with the overall taste. The final output is scored on a scale of 1-5 for each of the five basic tastes sensed by the human gustatory system, which are saltiness, sourness, sweetness, bitterness, and umami. The constructed network estimated the intensity of the basic tastes of 30 drink varieties with an average relative error of 7.0% compared to the human scores. The network could also estimate the variance in the human sensory perception. Moreover, the sensor successfully predicted the interactions of tastes such as suppression of bitterness by sweetness and enhancement of umami by saltiness, which are illusions sensed by the human gustatory system. With these abilities, the novel taste sensor can be considered as a quantitative yet humanlike sensor with a great potential for practical applications. PMID- 16351138 TI - Laser-induced acoustic desorption/fourier transform ion cyclotron resonance mass spectrometry for petroleum distillate analysis. AB - Laser-induced acoustic desorption (LIAD) coupled with a 3-T Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR) allows the simultaneous analysis of both the nonpolar and polar components in petroleum distillates. The LIAD/FT ICR method was validated by examining model compounds representative of the various classes of polar and nonpolar hydrocarbons commonly found in petroleum. LIAD successfully desorbs all the compounds as intact neutral molecules into the FT-ICR. Electron ionization (EI) at low energies (10 eV) and chemical ionization using cyclopentadienyl cobalt radical cation (CpCo*+) were employed to ionize the desorbed molecules. The EI experiments lead to extensive fragmentation of many of the hydrocarbon compounds studied. However, the CpCo*+ ion ionizes all the hydrocarbon compounds by producing only pseudomolecular ions without other fragmentation, with the exception of one compound (*CH3 loss occurs). Examination of two different petroleum distillate samples revealed hundreds of compounds. The most abundant components have an even molecular weight; i.e., they are likely to contain no (or possibly an even number of) nitrogen atoms. PMID- 16351139 TI - Using an oxide nanoarchitecture to make or break a proton wire. AB - We report that long-range proton diffusion (>0.3 mm) is generated in monolithic ultraporous manganese oxide nanoarchitectures upon exposure to gas-phase water. The sol-gel-derived ambigel nanoarchitectures, with bicontinuous networks of covalently bonded nanoscale solid and through-connected mesopores, exhibit conductometric sensitivity to humidity as established by impedance spectroscopy. The spectra contain a Warburg feature from which the concentration and diffusion length of the protonic charge carriers are determined. Water adsorbs conformally onto the architecture's continuous solid network in equilibrium with atmospheric humidity to create a continuous water sheath that acts as a 3-D proton wire. As a result, monolithic manganese oxide ambigels exhibit an equilibrium conductometric response to humidity that is 14 times greater than that of previous reports for electrolytic manganese oxide. A packed bed of 1-10-microm ambigel particulates in physical contact with one another, each with the same nanoscale morphology as the monolithic nanoarchitecture, also support long-range proton diffusion; however, the sensitivity to humidity is four times lower than the monolithic form due to restricted proton transport between adjacent particulates. Films composed of 0.3 12-microm ambigel particulates supported on interdigitated array electrodes with 20-microm electrode spacing express finite-diffusion behavior due to the short distance between the contact electrodes and have a conductometric sensitivity to humidity comparable to electrolytic MnO2 and 17 times lower than the monolithic ambigel. These results suggest that controlling the nature of the porous and solid phases in a nanoarchitecture provides a mechanism to limit interference from condensed water in conductometric gas-phase sensors. In addition, continuous monolithic architectures should improve electrochemical performance in devices where efficient long-range transport of protons or other ions is critical. PMID- 16351140 TI - Microchip separations in reduced-gravity and hypergravity environments. AB - Microfabricated fluidics technology, e.g., lab-on-a-chip devices, offers many attractive features for performing chemistry and biochemistry on space-based platforms. We have constructed a portable, battery-operated microfluidic platform that was tested under reduced gravity and hypergravity conditions that would be experienced in space flight and launch. This device consisted of a microchip, microchip holder, two 0-8-kV high-voltage power supplies, a high-voltage switch, a solid-state diode-pumped green laser, an optical train, a channel photomultiplier, and an inertial mass measurement unit all under the control of a laptop computer and powered by 10 D-cell alkaline batteries. The unit was tested on NASA's reduced gravity research aircraft at gravity levels that are relevant to NASA's intended use of bioreporter-based microchips for environmental monitoring of space and planetary environments on manned and unmanned spacecraft. Over the course of two flights, 834 fast electrophoretic separations of four amino acids were performed under a variety of gravitational environments including zero-g, Martian-g, lunar-g, and approximately 1.8-g. All separations were performed in less than 12 s and automatically analyzed. After correction with an internal migration standard, the migration time reproducibilities were all <1% relative standard deviation. PMID- 16351141 TI - Multiarray sensors with pattern recognition for the detection, classification, and differentiation of bacteria at subspecies and strain levels. AB - This work describes the integration of a fully autonomous electrochemical biosensor with pattern recognition techniques for the detection and classification of bacteria at subspecies and strain level. The system provides a continuous, real-time monitoring of bacteria activity upon exposure to antibiotics. The system utilizes 96-well-type electrodes array (DOX-dissolved oxygen sensor) with principal component analysis (PCA) for rapid and routine classification of different classes of bacteria and related strains. A representative sample of a section of the bacteria kingdom has been analyzed and classified using the proposed DOX-PCA system, including the following: Corynebacterium glutamicum, Micrococcus luteus, Staphylococcus epidermidis, Yersinia ruckeri, Escherichia adecarboxylata, Comamonas acidovorans, Alcaligenes odorans, Bacillus globigii, and three strains of Escherichia coli (K12, SM10, ATCC 25922). The new classification scheme is based on the hypothesis that, under identical experimental conditions, various bacteria consume oxygen at different rates and are affected in different ways by selected antibiotics. Thus, the response of the individual electrode in the array is indirectly altered, compared to that of cells growing on medium, by the addition of the antibiotic. By using three different antibiotics in separate wells, a unique fingerprint can be created for a specific bacterium. With the proposed DOX-PCA system, classification of bacteria was achieved at subspecies and strain level in real time. This study represents a basic research tool that may allow researchers to rapidly detect, quantify, and classify bacteria type at subspecies and strain levels. PMID- 16351142 TI - High-resolution TOF-SIMS imaging of eukaryotic cells preserved in a trehalose matrix. AB - A novel, trehalose-glycerol matrix was utilized to generate high-resolution, TOF SIMS images of macrophages and glial cells. Viable cells incubated in 50 mM trehalose, then lyophilized in a 50 mM trehalose, 10-15% (w/w) glycerol rinse, are preserved and chemically profiled. These experiments demonstrate the utility of the disaccharide matrix as an efficient, cost-effective alternative to cryogenics for SIMS and other ultrahigh-vacuum (UHV) analyses of biological species. Cellular processes on oligodendrocytes and astrocytes, 1-3 mum in width, were well resolved for cells in the trehalose-glycerol matrix. The viscous cell matrixes were fractured and analyzed at room temperature and maintained their three-dimensional integrity under UHV. Images have been generated with a Au primary ion source near the static limit of 10(12) ions/cm2. Though these nucleated cells do not remain viable after desiccation, TOF-SIMS imaging and subsequent rehydration reveals structural and morphological preservation. Eliminating the inherent obstacles associated with cryogenic analysis opens the door to greater utility of SIMS as a bioanalytical tool, such as lipid mapping of single cells in the nervous system. PMID- 16351143 TI - Detection of meningioma in dura mater by Raman spectroscopy. AB - Radical tumor resection is the treatment of choice for patients suffering from meningioma. However, recurrence of these tumors is a problem. Tumor recurrences are attributed to residual nests of meningioma within the regional dura. Therefore, complete removal of all tumor-infiltrated dura is important. Meningioma and normal dura were studied by Raman microspectroscopy to assess the possibility of developing an in vivo Raman method for guidance of meningioma resections. Pseudocolor Raman maps were constructed of cryosections of dura and meningioma, obtained from 20 patients. Comparison of these maps with histopathology enabled assignment of the spectra to either meningioma or dura. Large differences exist between the Raman spectra of dura and meningioma, because of the high collagen content of dura and the increased lipid content of tumors. A classification model for dura and tumor tissue based on linear discriminant analysis of Raman spectra yielded an accuracy of 100%. A first attempt was made to determine the minimum amount of meningioma in dura that is detectable by Raman spectroscopy. It is concluded that Raman spectra enable meningioma to be distinguished from dura, which makes Raman spectroscopy a viable candidate for guidance of surgical resection of meningioma. PMID- 16351144 TI - Scanning electrochemical microscopy. Hydrodynamics generated by the motion of a scanning tip and its consequences on the tip current. AB - The motion of a SECM tip above a substrate generates a flow of the surrounding fluid. Finite element calculations show that this flow is a simple linear-shear flow (Couette flow) for small tip-substrate separations and deviates from Couette's law at larger ones. The effect of fluid flow on the tip current response was determined numerically. Different mass-transfer regimes are observed depending on the insulating or conducting nature of the substrate, the tip speed (or fluid velocity), and the tip-substrate separation. Those observations are tested experimentally, and good agreement is obtained between numerical and experimental results. PMID- 16351145 TI - A scalable addressable positive-dielectrophoretic cell-sorting array. AB - We present the first known implementation of a passive, scalable architecture for trapping, imaging, and sorting individual microparticles, including cells, using a positive dielectrophoretic (p-DEP) trapping array. Our array-based technology enables "active coverslips" where, when scaled, many individually held cells can be sorted based upon imaged spatial or temporally variant characteristics. Our design incorporates a unique "ring-dot" p-DEP trap geometry organized in a row/column array format. This trap design, implemented in a two-level metal process, provides strong and highly spatially localized holding fields enabling single-cell capture for all traps in the array. We release individual trapped microparticles during sorting using a passive transistor-independent approach where we electrically ground the row and column electrodes associated with specific traps in the array. The demand for chip-to-world electrical connections in our arrays scales proportionally with the square root of the number of traps in a given array, delivering a substantial improvement over prior designs. We demonstrate capture, holding, and release operations with both beads and cells in small arrays of this new architecture. PMID- 16351146 TI - Isothermal DNA amplification coupled with DNA nanosphere-based colorimetric detection. AB - We present a simple, rapid method for detecting short DNA sequences that combines a novel isothermal amplification method (EXPAR) with visual, colorimetric readout based on aggregation of DNA-functionalized gold nanospheres. The reaction is initiated by a trigger oligonucleotide, synthetic in nature for this proof-of principle study, which is exponentially amplified at 55 degrees C and converted to a universal reporter oligonucleotide capable of bridging two sets of DNA functionalized gold nanospheres. This reaction provides >10(6)-fold amplification/conversion in under 5 min. When combined with a solution containing DNA nanospheres, the bridging reporter causes nanosphere aggregation. The resulting color change from red to dark purple or blue is enhanced through spotting the solution onto a C18 reversed-phase thin-layer chromatography plate. The reaction can easily be adapted for detection of different trigger oligonucleotides using the same set of DNA nanospheres. It permits detection of as low as 100 fM trigger oligonucleotide in under 10 min total assay time, with minimal reagent consumption and requirement for instrumentation. We expect that combining this simple, versatile assay with trigger generation from a genomic target DNA sequence of interest will be a powerful tool in the development of rapid and simple point-of-care molecular diagnostic applications. PMID- 16351147 TI - Microchip capillary electrophoresis with solid-state electrochemiluminescence detector. AB - We report microchip capillary electrophoresis (CE) coupling to a solid-state electrochemiluminescence (ECL) detector. The solid-state ECL detector was fabricated by immobilizing tris(2,2'-bipyridyl)ruthenium(II) (TBR) into an Eastman AQ55D-silica-carbon nanotube composite thin film on an indium tin oxide (ITO) electrode. After being made by a photolithographic method, the surface of the ITO electrode was coated with a thin composite film through a micromolding in capillary (MIMIC) technique using a poly(dimethylsiloxane) (PDMS) microchannel with the same pattern as an ITO electrode. Then the TBR was immobilized via ion exchange by immersing the ITO electrode containing the thin film in TBR aqueous solution. The whole system was built by reversibly sealing the TBR-modified ITO electrode plate with a PDMS layer containing electrophoresis microchannels. The results indicated that the present solid-state ECL detector displayed good durability and stability in the microchip CE-ECL system. Proline was selected to perform the microchip device with a limit of detection of 2 microM (S/N=3) and a linear range from 25 to 1000 microM. Compared with the CE-ECL of TBR in aqueous solution, while the CE microchip with solid-state ECL detector system gave the same sensitivity of analysis, a much lower TBR consumption and a high integration of the whole system were obtained. The present system was also used for medicine analysis. PMID- 16351148 TI - Sign constraints improve the detection of differences between complex spectral data sets: LC-IR as an example. AB - Spectroscopy is a fast and rich analytical tool. On many occasions, spectra are acquired of two or more sets of samples that differ only slightly. These data sets then need to be compared and analyzed, but sometimes it is difficult to find the differences. We present a simple and effective method that detects and extracts new spectral features in a spectrum coming from one set with respect to spectra of another set on the basis of the fact that these new spectral features are essentially positive quantities. The proposed procedure (i) characterizes the spectra of the reference set by a component model and (ii) uses asymmetric least squares (ASLS) to find differences with respect to this component model. It should be stressed that the method only focuses on new features and does not trace relative changes of spectral features that occur in both sets of spectra. A comparison is made with the conventional ordinary least squares (OLS) approach. Both methods (OLS and ASLS) are illustrated with simulations and are tested for size-exclusion chromatography with infrared detection (SEC-IR) of mixtures of polymer standards. Both methods are able to provide information about new spectral features. It is shown that the ASLS-based procedure yields the best recovery of new features in the simulations and in the SEC-IR experiments. Band positions and band shapes of new spectral features are better retrieved with the ASLS than with the OLS method, even those which could hardly be detected visually. Depending on the spectroscopic technique used, the ASLS-based method facilitates identification of the new chemical compounds. PMID- 16351149 TI - Reduction of tip-sample contact using dielectrophoretic force scanning probe microscopy. AB - Dielectrophoretic force microscopy is shown to allow for facile noncontact imaging of systems in aqueous media. Electrokinetic tip-sample forces were predicted from topography measurements of an interface and compared with experimental images. Correlation function and power spectral density analyses indicated that image feedback was maintained without mechanical contact using moderate potentials (e.g., approximately 18 nm off the surface for a 7-Vpp, 100 kHz waveform). The applied dielectrophoretic force and the corresponding increase in effective tip radius were predictably adjusted by changing the peak potential. PMID- 16351150 TI - Nanovolume kinase inhibition assay using a sol-gel-derived multicomponent microarray. AB - We report on the development of a new class of kinase microarrays based on the coimmobilization of both kinase and substrate components within a single pin printed sol-gel microarray element and the use of such arrays for nanovolume inhibition assays. We successfully immobilized the alpha-catalytic subunit of cAMP-dependent protein kinase (PKA) and the peptide substrate kemptide within sol gel-derived microarrays for the purpose of monitoring phosphorylation and inhibition. Using Pro-Q Diamond stain as an end-point indicator of phosphorylation, we demonstrate the selective detection of phosphoproteins over nonphosphorylated controls and the ability to detect phosphorylated proteins over a 500-fold concentration range. Limits of detection for the phosphoprotein beta casein were 7.5 pg, and the detectable signal remained linear up to 3.75 ng of protein per array spot. PKA is demonstrated to be active when coentrapped with two different substrates, and inhibition assays for PKA with the inhibitors H7 and H89 demonstrate the ability to detect kinase inhibition as well as derive IC50 plots from a single array using an overprinting method to deliver approximately 0.6 nL of reagent per array element, or a total of 72 nL of reagents to generate a full, 12-point IC50 curve in pentuplicate. PMID- 16351151 TI - Screening of target-specific stress-responsive genes for the development of cell based biosensors using a DNA microarray. AB - In this study, we describe a straightforward strategy to develop whole cell-based biosensors using fusions of the bacterial bioluminescence genes and the promoters from chemically responsive genes within Escherichia coli, in which chemical target-responsive genes were screened by using the information of gene expression data obtained from DNA microarray analysis. Paraquat was used as a model chemical to trigger gene expression changes of E. coli and to show the DNA microarray assisted development of whole cell-based biosensors. Gene expression data from the DNA microarray were obtained by time course analysis (10, 30, and 60 min) after exposure to paraquat. After clustering gene expression data obtained by time course analysis, a group of highly expressed genes over the all time courses could be classified. Within this group, three genes expressed highly for overall time points were selected and promoters of these genes were used as fusion partners with reporter genes, lux CDABE, to construct whole cell-based biosensors. The constructed biosensors recognized the presence of model inducer, paraquat, and structural analogue chemicals of paraquat with a high specificity, and the results were reconfirmed by using DNA microarray experiments for those structural analogues. This strategy to develop whole cell-based biosensors assisted by DNA microarray information should be useful in general for constructing chemical-specific or stress-specific biosensors with a high throughput manner. PMID- 16351152 TI - Calibration-free quantitative analysis of mRNA. AB - Here we introduce a method for accurate and sensitive quantitative analysis of mRNA, which does not require calibration with mRNA. The method uses a fluorescently labeled hybridization probe as a reference standard. It involves the following: (i) annealing mRNA to the excess of the fluorescently labeled ssDNA hybridization probe, (ii) separation of the mRNA-probe hybrid from the excess of the probe by gel-free capillary electrophoresis mediated by ssDNA binding protein, (iii) fluorescence detection of the hybrid and the excess probe, and (iv) quantification of mRNA using a simple algebraic formula. The method also overcomes a number of other limitations of conventional methods: the entire procedure currently takes only 2 h and accurately quantifies 10(5) copies of mRNA. With further improvements to the method, the procedure can be potentially shortened to 10 min, and the limit of quantification can be decreased to as few as 100 copies of mRNA. In this work, we prove the principle of the method by quantifying mRNA of green fluorescent protein in the matrix of total cellular RNA. The developed method is quantitative, simple, fast, and highly sensitive. It requires commercially available instrumentation only. The method will be an indispensable tool for molecular and cell biology studies. PMID- 16351153 TI - Continuous collection of soluble atmospheric particles with a wetted hydrophilic filter. AB - Approximately one-third of the area (14-mm diameter of a 25-mm diameter) of a 5 microm uniform pore size polycarbonate filter is continuously wetted by a 0.25 mL/min water mist. The water forms a continuous thin film on the filter and percolates through it. The flowing water substantially reduces the effective pore size of the filter. At the operational air sampling flow rate of 1.5 standard liters per minute, such a particle collector (PC) efficiently captures particles down to very small size. As determined by fluorescein-tagged NaCl aerosol generated by a vibrating orifice aerosol generator, the capture efficiency was 97.7+% for particle aerodynamic diameters ranging from 0.28 to 3.88 microm. Further, 55.3 and 80.3% of 25- and 100-nm (NH4)2SO4 particles generated by size classification with a differential mobility analyzer were respectively collected by the device. The PC is integrally coupled with a liquid collection reservoir. The liquid effluent from the wetted filter collector, bearing the soluble components of the aerosol, can be continuously collected or periodically withdrawn. The latter strategy permits the use of a robust syringe pump for the purpose. Coupled with a PM2.5 cyclone inlet and a membrane-based parallel plate denuder at the front end and an ion chromatograph at the back end, the PC readily operated for at least 4-week periods without filter replacement or any other maintenance. PMID- 16351154 TI - Expanded electrochemical capabilities of the electrospray ion source using porous flow-through electrodes as the upstream ground and emitter high-voltage contact. AB - Use of a porous flow-through electrode at the upstream ground contact or at both the upstream ground contact and the high-voltage emitter contact in an electrospray ion source was shown to provide for new types of electrochemical experiments utilizing only the electrochemistry inherent to electrospray. The normal stainless steel bore-through union serving as the upstream grounding point in a floated electrospray emitter system was replaced with a high surface area porous flow-through electrode assembly to achieve effective electrochemical reduction of analytes at this point in positive ion mode, and effective electrochemical oxidation of analytes in negative ion mode. This was demonstrated by the oxidation of 3,4-dihydroxybenzoic acid and reserpine in negative ion mode and by the reduction of thionine in positive ion mode. In the case of reversible oxidation (3,4-dihydroxybenzoic acid) and reduction (thionine) processes, partial rereduction and reoxidation of the products due to reaction with products generated by cathodic and anodic processes at the emitter were observed, respectively. By implementing two high surface area porous flow-through electrodes in the system, one as the upstream grounding point and the other as the emitter electrode, a multiple-step reaction scheme was achieved that included consecutive electrochemical reduction and oxidation reactions and a following chemical reaction as demonstrated by the hydroquinone tagging of an initially disulfide-linked peptide. PMID- 16351155 TI - Single-cell microarray for analyzing cellular response. AB - Detection of cellular response by measuring intracellular calcium, (Ca2+)i with Ca2+-dependent fluorescent dye are standard approaches to detect ligand stimulated cells and to study signaling through ligand/receptor interaction. We describe a single-cell microarray system to analyze cellular response of individual cells such as lymphocytes using microchamber array chips. The single cell microarray chip is made from polystyrene with over 30,000 microchambers, which can accommodate only single cells. Lymphocytes derived from mouse spleen or human blood were spread on the microarray, and over 80% of the microchambers achieved single-cell status. Stimulation of B-cells through antigen receptors on the microarray allowed us to detect activated B-cells by comparing the states of single B-cells before and after stimulation with antigen, which is disabled for flow cytometry. In addition, this novel method demonstrated retrieval of positive single B-cells from microchambers by a micromanipulator and achieved antibody DNA analysis. The system is suitable for high-throughput analysis of intracellular Ca2+ response at the single-cell level and is applicable to screen antigen specific lymphocytes for making specific monoclonal antibody. PMID- 16351156 TI - Microwave-accelerated metal-enhanced fluorescence: platform technology for ultrafast and ultrabright assays. AB - We describe an exciting assay platform technology that promises to fundamentally address two underlying physical constraints of modern assays and immunoassays, namely, assay sensitivity and rapidity. By combining the use of metal-enhanced fluorescence with low-power microwave heating, we can indeed significantly increase the sensitivity of surface assays as well as >95 % kinetically complete the assay within a few seconds. Subsequently, this new technology promises to fundamentally change the way we currently employ immunoassays in clinical medicine. This new model platform system can be potentially applied to many other important assays, such as to the clinical assessment of myoglobin, where both assay speed and sensitivity is paramount for the assessment and treatment of acute myocardial infarction. To demonstrate the utility of microwave-accelerated metal-enhanced fluorescence (MAMEF), we show that a simple protein-based assay system can be optically amplified approximately 10-fold by using silver nanostructures, while being kinetically complete in less than 20 s. This new platform approach is subsequently over 10-fold more sensitive and approximately 90 times faster than a control assay that operates both at room temperature and without the use of metal-enhanced fluorescence. Finally, we show that low-power heating by microwaves in our model system does not denature proteins, as evidenced by no protein structural changes, probed by fluorescence resonance energy transfer. PMID- 16351157 TI - Direct electrochemistry of myoglobin in titanate nanotubes film. AB - Titanate nanotubes (TNT) were proven to be efficient support matrixes for the immobilization of myoglobin (Mb). A comparative study was performed using the corresponding analogue, nanocrystalline anatase TiO2 (TNP). UV-visible absorption and FT-IR spectra show that Mb was not obviously denatured in TNT film in contrast to the significant denaturation of Mb in TNP film. Cyclic voltammetry and square wave voltammetry measurements were carried out using the Mb-TNT or Mb TNP cast film-covered basal plane pyrolytic graphite electrode. The Mb-TNT film gave a well-defined, nearly reversible redox couple with the apparent formal peak potential (Ep) of -0.239 V (vs Ag/AgCl) in pH 5.5 buffer, whereas a relatively smaller, quais-reversible redox pair with Ep of -0.263 V was observed for the Mb TNP film. The amounts of electroactive Mb in TNT film and TNP film were 15 and 10%, respectively. Moreover, the Mb-TNT film exerted facile direct electron transfer with the apparent heterogeneous electron-transfer rate constant (kET) of 86+/-7 s-1, almost 4 times the 22+/-5 s-1 value for the Mb-TNP membrane and higher than other Mb-entrapped films reported. Additionally, the Mb-TNT film demonstrates good electrocatalytic reduction of hydrogen peroxide with a detection limit of 0.6 microM, much lower than the 3.0 microM value for the Mb TNP electrode and other Mb-related film-modified electrodes reported so far. The Mb-TNT film exhibits higher peroxidase-like activity with the apparent Michaelis Menton constant (KM) of 140 microM, significantly lower than the 1300 microM value for the Mb-TNP film. The functional hydroxyl group and the surface charge as well as tubular morphology of TNT are important factors to stabilize the bound protein. PMID- 16351158 TI - Phosphorescence quenching of dyes adsorbed to silica thin-layer chromatography plates. AB - We describe photoluminescence (PL) quenching experiments by oxygen for a series of transition metal dyes adsorbed to commercial silica thin-layer chromatography plates. These TLC plates have been used by others as rapidly responding PL pressure sensors. The quenching kinetics show interesting differences from the behavior of the same dyes adsorbed to the more well-defined surfaces of SBA-15 mesoporous silica particles adsorbed to a thin layer-by-layer polymer film as reported in Chem. Mater. 2005, 17, 3160. The pore size and the pore size distribution are much larger for the TLC silica than for the mesoporous silica. On the TLC silica, the dye PL decay profiles show smaller deviations from an exponential form. One sees larger differences between the intensity and lifetime Stern-Volmer plots, and most surprisingly, the dyes on the surface of the TLC silica are about three times more sensitive to quenching by oxygen. PMID- 16351159 TI - Extraction and GC/MS analysis of the human blood plasma metabolome. AB - Analysis of the entire set of low molecular weight compounds (LMC), the metabolome, could provide deeper insights into mechanisms of disease and novel markers for diagnosis. In the investigation, we developed an extraction and derivatization protocol, using experimental design theory (design of experiment), for analyzing the human blood plasma metabolome by GC/MS. The protocol was optimized by evaluating the data for more than 500 resolved peaks using multivariate statistical tools including principal component analysis and partial least-squares projections to latent structures (PLS). The performance of five organic solvents (methanol, ethanol, acetonitrile, acetone, chloroform), singly and in combination, was investigated to optimize the LMC extraction. PLS analysis demonstrated that methanol extraction was particularly efficient and highly reproducible. The extraction and derivatization conditions were also optimized. Quantitative data for 32 endogenous compounds showed good precision and linearity. In addition, the determined amounts of eight selected compounds agreed well with analyses by independent methods in accredited laboratories, and most of the compounds could be detected at absolute levels of approximately 0.1 pmol injected, corresponding to plasma concentrations between 0.1 and 1 microM. The results suggest that the method could be usefully integrated into metabolomic studies for various purposes, e.g., for identifying biological markers related to diseases. PMID- 16351160 TI - High-performance SPME/AP MALDI system for high-throughput sampling and determination of peptides. AB - This paper presents the performance characteristics for a new multiplexed solid phase microextraction/atmospheric pressure matrix-assisted laser desorption/ionization (SPME/AP MALDI) source configuration for a hybrid quadrupole-linear ion trap instrument. The results demonstrate that thorough optimization of parameters such as SPME coating material, optics configurations, extraction solvents, and fiber capacity provides dramatic sensitivity improvements (>1000x) over previous reports in the literature. The multiplexed SPME plate is capable of simultaneous extraction from 16 different wells on a multiwell plate, eliminating the need for extensive sample preparation. Subfemtomole sensitivity is demonstrated for peptide standards and protein digests with run-run reproducibility ranging from approximately 13 to 31%. This high-performance SPME/AP MALDI system shows potential for high-throughput extraction from biological samples. PMID- 16351161 TI - Characterization and manipulation of the electroosmotic flow in porous anodic alumina membranes. AB - Porous anodic alumina membranes (PAAMs) have uniform and high-density nanopores, and the dimension and interval of the pores can be easily controlled by varying the anodization conditions. The application of PAAMs could widely impact the cost and efficiency of the liquid-based nanoscale separations. We report here the property of electroosmotic flow in PAAMs, which plays a significant role in the mass transport across these membranes that have charged pore surfaces. By controlling the solution pH and the magnitude and sign of the applied current, the mass transport through these nanoporous membranes can be spatially and temporally manipulated. The effects of electrosurface properties and electrolyte ionic strength on electroosmotic flow were studied. The anion incorporation and adsorption cause the variation of the electrosurface properties of PAAMs, which in turn influence the rate and direction of the mass transport. As compared to the membrane with fixed surface charge, this diversity makes it possible for the PAAMs to be used in various conditions. PMID- 16351162 TI - NMR analysis on microfluidic devices by remote detection. AB - We present a novel approach to perform high-sensitivity NMR imaging and spectroscopic analysis on microfluidic devices. The application of NMR, the most information-rich spectroscopic technique, to microfluidic devices remains a challenge because the inherently low sensitivity of NMR is aggravated by small fluid volumes leading to low NMR signal and geometric constraints resulting in poor efficiency for inductive detection. We address the latter by physically separating signal detection from encoding of information with remote detection. Thereby, we use a commercial imaging probe with sufficiently large diameter to encompass the entire device, enabling encoding of NMR information at any location on the chip. Because large-diameter coils are too insensitive for detection, we store the encoded information as longitudinal magnetization and flow it into the outlet capillary. There, we detect the signal with optimal sensitivity, using a solenoidal microcoil, and reconstruct the information encoded in the fluid. We present a generally applicable design for a detection-only microcoil probe that can be inserted into the bore of a commercial imaging probe. Using hyperpolarized 129Xe gas, we show that this probe enables sensitive reconstruction of NMR spectroscopic information encoded by the large imaging probe while keeping the flexibility of a large coil. PMID- 16351163 TI - In-a-day electrochemical detection of coliforms in drinking water using a tyrosinase composite biosensor. AB - A rapid method for the detection of fecal contamination in water based on the use of a tyrosinase composite biosensor for improved amperometric detection of beta galactosidase activity is reported. The method relies on the detection of phenol released after the hydrolysis of phenyl beta-D-galactopyranoside (PG) by beta galactosidase. Under the optimized PG concentration and pH (4.0) values, a detection limit of 1.2x10(-3) unit of beta-galactosidase/mL-1 was obtained. The capability of the sensor for the detection of Escherichia coli was evaluated using polymyxin B sulfate to allow permeabilization of the bacteria membrane. A detection limit of 1x10(6) cfu of E. coli mL-1 was obtained with no preconcentration or pre-enrichment steps. To improve the analytical characteristics for bacteria detection, the processes involving galactosidase induction during incubation and membrane permeabilization were optimized. Using 0.25 mM isopropyl beta-D-thiogalactopyranoside for the enzyme activity induction, and 10 microg mL-1 polymyxin B sulfate as permeabilizer agent, it was possible to detect bacteria concentrations as low as 10 cfu mL-1 after 5 h of enrichment. The possibility of detecting E. coli at the required levels for drinking water quality assessment (1 cfu/100 mL) is demonstrated, the time of analysis being shorter than 6.5 h and involving a simple methodology. PMID- 16351164 TI - Kinetic calibration for automated headspace liquid-phase microextraction. AB - The kinetics of the absorption and desorption of analytes for headspace liquid phase microextraction (HS-LPME) were studied. It was found that the desorption of analytes from the extraction phase into the sample matrix is isotropic to the absorption of the analytes from the sample matrix into the extraction phase under the same conditions. This therefore allows for the calibration of absorption using desorption. Calibration was accomplished by exposing the extraction phase, which contained a standard, to the sample matrix. The information from the desorption of the standard, such as time constant a, could be directly used to estimate the concentration of the target analyte in the sample matrix. This new kinetic calibration method for headspace LPME was successfully used to correct the matrix effects in the BTEX analysis of an orange juice sample. In this study, the headspace LPME techniques were successfully fully automated, for both static and dynamic methods, with the CTC CombiPal autosampler. All operations of headspace LPME, including sample transfer and agitation, filling of extraction solvent, exposing the solvent in the headspace, withdrawing the solvent to syringe and introducing the extraction phase into injector, were autoperformed by the CTC autosampler. The fully automated headspace LPME technique is more convenient and improved the precision and sensitivity of the method. This automated dynamic headspace LPME technique can be also used to obtain the distribution coefficient between the sample matrix (aqueous or another solution) and the extraction phase (1-octanol or another solvent). The distribution coefficient between 1-octanol and orange juice, at 25 degrees C, was obtained with this technique. PMID- 16351165 TI - Combination of analyte protectants to overcome matrix effects in routine GC analysis of pesticide residues in food matrixes. AB - Analyte protectants were previously defined as compounds that strongly interact with active sites in the gas chromatographic (GC) system, thus decreasing degradation, adsorption, or both of coinjected analytes. In this study, we evaluated various combinations of promising analyte protectants for the volatility range of GC-amenable pesticides using GC/quadrupole mass spectrometry (MS) and 1-microL hot splitless injection for sample introduction. A mixture of ethylglycerol, gulonolactone, and sorbitol (at 10, 1, and 1 mg/mL, respectively, in the injected samples) was found to be the most effective in minimizing losses of susceptible analytes and significantly improving their peak shapes (due to reduction of peak tailing). When added to final sample extracts and matrix-free calibration standards alike, these analyte protectants induced a similar response enhancement in both instances, resulting in effective equalization of the matrix induced response enhancement effect even after a large number of fruit and vegetable extract injections. As compared to matrix-matched standardization, the analyte protectant approach offers a more convenient solution to the problems associated with calibration in routine GC/MS analysis of pesticide residues and possibly other susceptible analyte types in diverse samples. Moreover, the use of analyte protectants also substantially reduced another adverse matrix-related effect caused by gradual build-up of nonvolatile matrix components in the GC system, thus improving ruggedness and, consequently, reducing need for frequent maintenance. PMID- 16351166 TI - Adsorption of duplex DNA on mesoporous silicas: possibility of inclusion of DNA into their mesopores. AB - It is well known that a silica surface cannot adsorb duplex DNA in common aqueous solution (not chaotropic solution) because of the electrostatic repulsion of the silica surface and polyanionic DNA. However, we recently found that when duplex DNA in phosphoric acid form (or in acidic solution) was used, DNA was successfully adsorbed into mesoporous silicas even in low-salt aqueous solution. The adsorption behaviors of DNA into mesoporous silicas were influenced by the pore diameter sizes. Mesoporous silicas with 2.80- or 3.82-nm peak pore diameters adsorbed DNA the best in diluted NaCl solution. Formation of the hydrogen bond between P(O)OH groups in DNA and adsorbed water, SiOH groups, or both on silica surfaces is regarded as a main factor in this adsorption. The coincidence of the pore sizes and DNA diameter realizes this unique adsorption promoted by the effect of encompassing DNA with the inner surface of mesoporous silica. Although there is no clear direct evidence for including duplex DNA in the mesopores yet, this adsorption technique is expected to provide a new tool for DNA science, because DNA in the pore size 2-5 nm in diameter has to be in unusual disentangled thread form. PMID- 16351167 TI - Simultaneous observation of enzyme surface diffusion and surface reaction using microfluidic patterning of substrate surfaces. AB - We present a study of the simultaneous observation of protease reaction and surface diffusion as the enzyme interacts with a model substrate surface. We use micro-fluidic patterning to decorate a bovine serum albumin substrate surface with stripes of adsorbed enzyme in the absence of physical barriers. Spreading of the enzyme from the initial striped region indicates surface diffusion, while removal of the substrate provides a measure of reactivity. Microfluidic patterning provides a means to determine the relative importance of enzyme adsorption, surface diffusion, and reaction on the rate of substrate removal. PMID- 16351168 TI - Profiling an electrospray plume using surface-enhanced Raman spectroscopy. AB - We report the use of silver nanoparticles to obtain surface-enhanced Raman spectra of Crystal Violet in an electrospray plume. Surface enhancement allowed detection at low concentrations with the high specificity afforded by vibrational spectroscopy. SERS spectra were used to obtain an axial concentration profile closely matching that obtained in previous fluorescence experiments. SERS can provide more analyte structural information than has been obtainable from fluorescence studies of the plume. PMID- 16351169 TI - Aflatoxin screening by MALDI-TOF mass spectrometry. AB - Efficient detection of aflatoxins B1, B2, G1, and G2 has been performed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry using a UV absorbing ionic liquid matrix to obtain "matrix-free" mass spectra and addition of NaCl to enhance sensitivity via Na+ cationization. Using ionic alpha-cyano-4 hydroxycinnamic acid (Et3N-alpha-CHCA) as the matrix, matrix-free mass spectra in the m/z range of interest are acquired, and the B1, B2, G1, and G2 aflatoxins are readily detected with an LOD as low as 50 fmol. The technique is fast, requires little sample preparation and no derivatization or chromatographic separation, and seems therefore to be suitable for high-throughput aflatoxin screening. It should be easily extended to other micotoxins and provide an attractive technique to control the quality of major crops subjected to huge world commercial trades such as peanuts, corn, and rice as well as to monitor bioterrorism threats by micotoxin poisoning. PMID- 16351170 TI - Rational attachment of synthetic triptycene orthoquinone onto carbon nanotubes for electrocatalysis and sensitive detection of thiols. AB - This study demonstrates a novel electrochemical method for sensitive determination of biological thiols including homocysteine, cysteine, and glutathione based on rational functionalization of single-walled carbon nanotubes (SWNTs) with synthetic triptycene orthoquinone (TOQ). Unlike previous strategies used for the functionalization of the carbon nanotubes to fabricate new kind of electrochemically functional nanostructures, the method demonstrated here is essentially based on understanding of the redox properties inherent in the SWNTs themselves. It is demonstrated that the electrochemical oxidation of the thiols at the SWNT-modified electrode is redox-mediated by the quinone-like functional groups at the tube ends and that the low density of such functional groups leads to a follow-up oxidation of the thiols at a more positive potential at the electrode. To mimic the redox properties of the SWNTs and thus to increase the catalytic sites onto the SWNTs, we rationally choose the synthetic TOQ and attach such a compound onto the SWNTs. As a result, it is found that the rational attachment of TOQ onto the SWNTs substantially results in a sufficient electrocatalysis toward the thiols at a low potential of 0.0 V with enhanced sensitivities (i.e., almost by a factor of 10-fold) for the determination of such kind of species in relative to those at the SWNT-modified electrode. The high sensitivity and the good stability as well as the high reproducibility of the TOQ/SWNT-modified electrodes substantially make them very useful for reliable and durable determination of the biological thiols. PMID- 16351171 TI - Method for effective immobilization of Ru(bpy)(3)2+ on an electrode surface for solid-state electrochemiluminescene detection. AB - A novel method for effective immobilization of Ru(bpy)3(2+) on an electrode surface is developed. The whole process involves two steps: the electrostatic interactions between citrate-capped gold nanoparticles (AuNPs) and Ru(bpy)3Cl2 in aqueous medium were used to fabricate Ru(bpy)(3)2+-AuNP aggregates (Ru-AuNPs) first, and then the Au-S interactions between as-formed Ru-AuNPs and sulfhydryl groups were used to effectively immobilize the Ru-AuNPs on a sulfhydryl-derivated indium tin oxide (ITO) electrode surface. As-prepared ITO electrode shows excellent stability, and the ECL active species Ru(bpy)3(2+) contained therein exhibit excellent ECL behaviors. PMID- 16351172 TI - Electrospray from nanostructured tungsten oxide surfaces with ultralow sample volume. AB - This study demonstrated the feasibility of performing protein analysis with ultralow sample volume by combining a tungsten oxide nanowire (TON) fiber with a miniaturized electrospray ionization interface. An increase in wettability of the tugsten surface after growing randomly oriented TON on its surface allows strong adhesion of approximately 50 nL of the methanol solution at its tip. Under the influence of a high electric field, electrospray from a Taylor cone on the adhered methanol solution was observed and the multiply charged ions of protein molecules predissolved in the solution were detected. PMID- 16351173 TI - A bio-bar-code assay based upon dithiothreitol-induced oligonucleotide release. AB - The recently developed bio-bar-code assay for the PCR-less detection of protein and nucleic acid targets has been shown to be extraordinarily sensitive, exhibiting low attomolar sensitivity for protein targets and high zeptomolar sensitivity for nucleic acid targets. In the case of DNA detection, the original assay relies on three distinct oligonucleotide strands on a single nanoparticle for target identification and signal amplification. Herein, we report the development of a new nanoparticle probe that can be used in the bio-bar-code assay, which requires only one thiolated oligonucleotide strand. This new assay relies on the ability to liberate the adsorbed thiolated oligonucleotides from the gold nanoparticle surface with dithiothreitol (DTT), which simplifies the assay and increases its quantitative capabilities. The utility of this new DTT based system is demonstrated by detecting a mock mRNA target using both fluorescent and scanometric assay readouts. When the scanometric readout is used, the sensitivity of the assay is 7 aM and quantification can be accomplished over the low-attomolar to the mid-femtomolar concentration range. PMID- 16351174 TI - High-throughput identification of in-gel digested proteins by rapid, isocratic HPLC/MS/MS. AB - The high sensitivity and accuracy of mass spectrometry for identifying proteins has led to an explosive expansion of proteomics research, necessitating rapid procedures for HPLC/MS/MS analysis. Current HPLC/MS/MS analysis usually relies on elution of peptides from the HPLC column with a gradient that takes a total of 45 70 min for each cycle, limiting sample throughput and the speed of protein identification. Here we report a simple method for high-throughput protein identification, using isocratic, either methanol- or acetonitrile-based buffer systems, HPLC elution into an LTQ mass spectrometer. This procedure allows each cycle of highly sensitive HPLC/MS/MS analysis to be completed in 5 min, thus boosting the efficiency of HPLC/MS/MS analysis 9-14-fold. Using this method, each operator can acquire HPLC/MS/MS data for 96 in-gel proteolytic digests in one 8-h working day. The method can easily be implemented in any laboratory with an LTQ mass spectrometer. This protocol should find wide application in mass spectrometry laboratories that require high-throughput analysis but are limited by inefficient use of machine time. PMID- 16351175 TI - Key roles for chain flexibility in block copolymer membranes that contain pores or make tubes. AB - Block copolymer amphiphiles that self-assemble into membranes present robust and functionalizable alternatives to biological assemblies. Coarse-grained molecular dynamics shows that thick bilayers of A-B copolymers accommodate protein-like channels and also tend to regulate transport. This occurs as flexible, hydrophilic A chains insert into the pore and obstruct water entry. A-B-A triblocks that exploit "hairpin" and "straight" conformations also show assembly into novel nanotubules and further highlight the key roles for chain flexibility in biomimetic block copolymer assemblies. PMID- 16351176 TI - Three-dimensional control of protein patterning in microfabricated devices. AB - We present a new procedure that allows for the controlled patterning of functional proteins on the sidewalls of three-dimensional microfabricated chambers. We build microchambers with walls containing gold areas of a defined submicrometer size, to which proteins can be bound specifically, via thiol chemistry. We tested our system by observing the gliding of microtubules along motor-coated microchamber sidewalls and find that introducing an intermediate multilayer of proteins greatly enhances motor protein activity. This combination of spatial confinement and three-dimensional patterning of protein activity opens up new possibilities for the in vitro study of cellular processes, as well as for the development of guided transport mechanisms in nanodevices. PMID- 16351177 TI - Length-dependent extraction of single-walled carbon nanotubes. AB - A two-phase liquid-liquid extraction process is presented which is capable of extracting water-soluble single-walled carbon nanotubes into an organic phase. The extraction utilizes electrostatic interactions between a common phase transfer agent and the sidewall functional groups on the nanotubes. Large length dependent van der Waals forces for nanotubes allow the ability to control the length of nanotubes extracted into the organic phase as demonstrated by atomic force microscopy. PMID- 16351178 TI - Temperature dependence of excitonic radiative decay in CdSe quantum dots: the role of surface hole traps. AB - Using atomistic, semiempirical pseudopotential calculations, we show that if one assumes the simplest form of a surface state in a CdSe nanocrystal--an unpassivated surface anion site--one can explain theoretically several puzzling aspects regarding the observed temperature dependence of the radiative decay of excitons. In particular, our calculations show that the presence of surface states leads to a mixing of the dark and bright exciton states, resulting in a decrease of 3 orders of magnitude of the dark-exciton radiative lifetime. This result explains the persistence of the zero-phonon emission line at low temperature, for which thermal population of higher-energy bright-exciton states is negligible. Thus, we suggest that surface states are the controlling factor of dark-exciton radiative recombination in currently synthesized colloidal CdSe nanocrystals. PMID- 16351179 TI - Reversible bistable switching in nanoscale thiol-substituted oligoaniline molecular junctions. AB - Single molecular monolayers of oligoaniline dimers were integrated into sub-40-nm diameter metal nanowires to form in-wire molecular junctions. These junctions exhibited reproducible room temperature bistable switching with zero-bias high- to low-current state conductance ratios of up to 50, switching threshold voltages of approximately +/-1.5 V, and no measurable decay in the high-state current over 22 h. Such switching was not observed in similarly fabricated saturated dodecane (C12) or conjugated oligo(phenylene ethynylene) (OPE) molecular junctions. The low- and high-state current versus voltage was independent of temperature (10-300 K), suggesting that the dominant transport mechanism in these junctions is coherent tunneling. Inelastic electron tunneling spectra collected at 10 K show a change in the vibrational modes of the oligoaniline dimers when the junctions are switched from the low- to the high-current state. The results of these measurements suggest that the switching behavior is an inherent molecular feature that can be attributed to the oligoaniline dimer molecules that form the junction. PMID- 16351180 TI - Nonequilibrium steady state of a nanometric biochemical system: determining the thermodynamic driving force from single enzyme turnover time traces. AB - A single enzyme molecule in a living cell is a nanometric system that catalyzes biochemical reactions in a nonequilibrium steady-state condition. The chemical driving force, Deltamu, is an important thermodynamic quantity that determines the extent to which the reaction system is away from equilibrium. Here we show that Deltamu for an enzymatic reaction in situ can be determined from the nonequilibrium time traces for enzymatic turnovers of individual enzyme molecules, which can now be recorded experimentally by single-molecule techniques. Three different Deltamu estimators are presented from principles of nonequilibrium statistical mechanics: fluctuation theorem, Kawasaki identity, and fluctuation dissipation theorem, respectively. In particular, a maximum likelihood estimation method of Deltamu has been derived based on fluctuation theorem. The statistical precisions of these three Deltamu estimators are analyzed and compared for experimental time traces with finite lengths. PMID- 16351181 TI - Enhanced third-order nonlinear optical properties in dendrimer-metal nanocomposites. AB - Nonlinear optical (NLO) and time-resolved fluorescence spectroscopic measurements of dendrimer-metal nanocomposites (DNCs) are reported. Third-order NLO effects were investigated by degenerate four-wave mixing (DFWM) measurements for DNCs and DNCs incorporated into thin polymeric films. The results show an enhancement of the third-order nonlinear susceptibility for the chromophore-functionalized dendrimer-metal nanocomposites. Investigations of the mechanism of the enhanced NLO effect suggested a strong contribution because of the metal's local field. These results show the potential usefulness of dendrimer-metal nanocomposites for nonlinear optical effects and biophotonic applications. PMID- 16351182 TI - Cantilever sensor for nanomechanical detection of specific protein conformations. AB - A fast, label-free, and multiplexed method based on piezoresistive cantilevers is reported for the detection of specific protein conformations at the nanoscale level. The ligand-binding domain of the human oestrogen receptor (ERalpha-LBD) is used as the experimental model system, and ERalpha-LBD with or without oestradiol (E2) is detected using the conformation-specific peptides alpha/betaI (Ser-Ser Asn-His-Gln-Ser-Ser-Arg-Leu-Ile-Glu-Leu-Leu-Ser-Arg, which recognizes E2-bound ER) and alpha/betaII (Ser-Ala-Pro-Arg-Ala-Thr-Ile-Ser-His-Tyr-Leu-Met-Gly-Gly, which recognizes E2-free ER). Target-specific signals are obtained in situ at protein concentrations of 2.5-20 nM. The in-build electrical readout of the piezoresistive cantilevers provides a convenient alternative to the conventional optical detection, and the presented method offers the possibility of detecting protein conformational changes using miniaturized microarrays. PMID- 16351183 TI - Determination of the bending modulus of an individual multiwall carbon nanotube using an electric harmonic detection of resonance technique. AB - We report a new method of measuring the amplitude and phase of oscillations of individual multiwall carbon nanotubes (MWNTs). As in many other experiments, we excite the oscillations electrostatically, but we show that we can detect the amplitude and phase of the resulting oscillation electrically. As an example, we present measurements of the fundamental and first two overtones of the diving board resonance of a MWNT at 0.339, 2.42, and 5.31 MHz in ambient conditions. The corresponding quality factors were 67, 36, and 25. PMID- 16351184 TI - Evidence of sequential lift in growth of aligned multiwalled carbon nanotube multilayers. AB - We synthesized aligned multiwalled carbon nanotube multilayers by aerosol assisted catalytic chemical vapor deposition through sequential injections of aerosols containing both carbon and catalyst precursors. Each sequence was traced by a specific duration or precursor mixture, with the carbon source being possibly enriched in (13)C isotope labels. We discovered that any sequence involved the growth of a new layer on the substrate surface, under any pre existing one by lifting it up, giving definitive evidence of a base-growth mechanism. PMID- 16351185 TI - Sequence-encoded self-assembly of multiple-nanocomponent arrays by 2D DNA scaffolding. AB - Regular 2D arrays of multiple types of nanocomponents were constructed by self assembly to DNA scaffolding with alternating rows of sequence-encoded hybridization sites. Different-sized Au particles coated with DNA complementary to one of the sites were bound to the scaffolding, producing alternating rows of the two nanocomponents with a 32-nm inter-row spacing. These results demonstrate the potential for using DNA to self-assemble complex arrays of components with nanometer-scale precision. PMID- 16351186 TI - Improving the ductility of nanocrystalline bcc metals. AB - Nanocrystalline metals present extremely high yield strengths but limited ductility. Using atomistic simulations, we show that the fracture resistance of bcc nanocrystalline materials increases with decreasing grain size below a critical grain size. There appears to be a "most brittle" grain size corresponding to the "strongest size" that has been postulated. Impurities that strengthen the grain boundaries can improve ductility significantly for the relatively larger grain sizes, whereas ductility decreases for the smallest grain sizes. PMID- 16351187 TI - Solution-processed zinc oxide field-effect transistors based on self-assembly of colloidal nanorods. AB - Colloidal zinc oxide (ZnO) nanocrystals are attractive candidates for a low temperature and solution-processible semiconductor for high-performance thin-film field-effect transistors (TFTs). Here we show that by controlling the shape of the nanocrystals from spheres to rods the semiconducting properties of spin coated ZnO films can be much improved as a result of increasing particle size and self-alignment of the nanorods along the substrate. Postdeposition hydrothermal growth in an aqueous zinc ion solution has been found to further enhance grain size and connectivity and improve device performance. TFT devices made from 65-nm long and 10-nm-wide nanorods deposited by spin coating have been fabricated at moderate temperatures of 230 degrees C with mobilities of 0.61 cm(2)V(-1)s(-1) and on/off ratios of 3 x 10(5) after postdeposition growth, which is comparable to the characteristics of TFTs fabricated by traditional sputtering methods. PMID- 16351188 TI - Capacitance and conductance of single-walled carbon nanotubes in the presence of chemical vapors. AB - Simultaneous conductance and capacitance measurements on a single-walled carbon nanotube (SWNT) network are used to extract an intrinsic property of molecular adsorbates. Adsorbates from dilute chemical vapors produce a rapid response in both the capacitance and the conductance of the SWNT network. These responses are caused by a combination of two distinct physiochemical properties of the adsorbates: charge transfer and polarizability. We find that the ratio of the conductance response to the capacitance response is a concentration-independent intrinsic property of a chemical vapor that can assist in its identification. PMID- 16351189 TI - Protein crystals as scanned probes for recognition atomic force microscopy. AB - Lysozyme crystal growth has been localized at the tip of a conventional silicon nitride cantilever through seeded nucleation. After cross-linking with glutaraldehyde, lysozyme protein crystal tips image gold nanoparticles and grating standards with a resolution comparable to that of conventional tips. Force spectra between the lysozyme crystal tips and surfaces covered with antilysozyme reveal an adhesion force that drops significantly upon blocking with free lysozyme, thus confirming that lysozyme crystal tips can detect molecular recognition interactions. PMID- 16351190 TI - Field-effect transistors based on self-organized molecular nanostripes. AB - Charge transport properties in organic semiconductors depend strongly on molecular order. Here we demonstrate field-effect transistors where drain current flows through a precisely defined array of nanostripes made of crystalline and highly ordered molecules. The molecular stripes are fabricated across the channel of the transistor by a stamp-assisted deposition of the molecular semiconductors from a solution. As the solvent evaporates, the capillary forces drive the solution to form menisci under the stamp protrusions. The solute precipitates only in the regions where the solution is confined by the menisci once the critical concentration is reached and self-organizes into molecularly ordered stripes 100-200 nm wide and a few monolayers high. The charge mobility measured along the stripes is 2 orders of magnitude larger than the values measured for spin-coated thin films. PMID- 16351191 TI - Nanotechnology-based photodynamic therapy for neovascular disease using a supramolecular nanocarrier loaded with a dendritic photosensitizer. AB - Photodynamic therapy (PDT) for exudative age-related macular degeneration (AMD) was evaluated using a supramolecular nanomedical device, that is, a novel dendritic photosensitizer (DP) encapsulated by a polymeric micelle formulation. The characteristic dendritic structure of the DP prevents aggregation of its core sensitizer, thereby inducing a highly effective photochemical reaction. With its highly selective accumulation on choroidal neovascularization (CNV) lesions, this treatment resulted in a remarkably efficacious CNV occlusion with minimal unfavorable phototoxicity. PMID- 16351192 TI - Self-organized growth of Si/Silica/Er2Si2O7 core-shell nanowire heterostructures and their luminescence. AB - Self-organized Si-Er heterostructure nanowires showed promising 1.54 microm Er(3+) optical activity. Si nanowires of about 120-nm diameter were grown vertically on Si substrates by the vapor-liquid-solid mechanism in an Si-Er-Cl H(2) system using an Au catalyst. Meanwhile, a single-crystalline Er(2)Si(2)O(7) shell sandwiched between nanometer-thin amorphous silica shells was self organized on the surface of Si nanowires. The nanometer-thin heterostructure shells make it possible to observe a carrier-mediated 1.53 microm Er(3+) photoluminescence spectrum consisting of a series of very sharp peaks. The Er(3+) spectrum and intensity showed absolutely no change as the temperature was increased from 25 to 300 K. The luminescence lifetime at room temperature was found to be 70 micros. The self-organized Si nanowires show great potential as the material basis for developing an Si-based Er light source. PMID- 16351193 TI - A novel silicon nanotips antireflection surface for the micro Sun sensor. AB - We have developed a new technique to fabricate an antireflection surface using silicon nanotips for use on a micro Sun sensor for Mars rovers. We have achieved randomly distributed nanotips of radii spanning from 20 to 100 nm and aspect ratio of approximately 200 using a two-step dry etching process. The 30 degrees specular reflectance at the target wavelength of 1 microm is only about 0.09%, nearly 3 orders of magnitude lower than that of bare silicon, and the hemispherical reflectance is approximately 8%. When the density and aspect ratio of these nanotips are changed, a change in reflectance is demonstrated. When surfaces are covered with these nanotips, the critical problem of ghost images that are caused by multiple internal reflections in a micro Sun sensor was solved. PMID- 16351194 TI - Resonant coupling between surface vibrations and electronic states in silicon nanocrystals at the strong confinement regime. AB - A striking correlation between infrared photoinduced absorption spectra and the photoluminescence from silicon nanocrystals indicates that quantized electronic sublevels of the nanocrystals are resonantly coupled to surface vibrational modes via a polarization field produced by coherent longitudinal polar vibrations. Our experimental results and model support the assumption that the mechanism responsible for the efficient photoluminescence from silicon nanocrystals should be assigned to inhibition of nonradiative channels rather than enhancement of radiative channels. PMID- 16351196 TI - Dual Raman features of double coaxial carbon nanotubes with N-doped and B-doped multiwalls. AB - Double coaxial carbon nanotubes with nitrogen (N)-doped and boron (B)-doped multiwalls possess composite Raman characteristics, originating not only from the outer N-doped but also from inner B-doped layers. Both N and B dopings result in substantial shifts of the characteristic D band and G band of sp(2) carbon constituting nanotube walls but in different ways. The downshift of the G band is correlated with the decreases of electrical resistivity of carbon nanotubes regardless of N or B doping. PMID- 16351195 TI - Molecular characterization of the cytotoxic mechanism of multiwall carbon nanotubes and nano-onions on human skin fibroblast. AB - The increasing use of nanotechnology in consumer products and medical applications underlies the importance of understanding its potential toxic effects to people and the environment. Although both fullerene and carbon nanotubes have been demonstrated to accumulate to cytotoxic levels within organs of various animal models and cell types and carbon nanomaterials have been exploited for cancer therapies, the molecular and cellular mechanisms for cytotoxicity of this class of nanomaterial are not yet fully apparent. To address this question, we have performed whole genome expression array analysis and high content image analysis based phenotypic measurements on human skin fibroblast cell populations exposed to multiwall carbon nano-onions (MWCNOs) and multiwall carbon nanotubes (MWCNTs). Here we demonstrate that exposing cells to MWCNOs and MWCNTs at cytotoxic doses induces cell cycle arrest and increases apoptosis/necrosis. Expression array analysis indicates that multiple cellular pathways are perturbed after exposure to these nanomaterials at these doses, with material-specific toxigenomic profiles observed. Moreover, there are also distinct qualitative and quantitative differences in gene expression profiles, with each material at different dosage levels (6 and 0.6 microg/mL for MWCNO and 0.6 and 0.06 microg/mL for MWCNT). MWCNO and MWCNT exposure activates genes involved in cellular transport, metabolism, cell cycle regulation, and stress response. MWCNTs induce genes indicative of a strong immune and inflammatory response within skin fibroblasts, while MWCNO changes are concentrated in genes induced in response to external stimuli. Promoter analysis of the microarray results demonstrate that interferon and p38/ERK-MAPK cascades are critical pathway components in the induced signal transduction contributing to the more adverse effects observed upon exposure to MWCNTs as compared to MWCNOs. PMID- 16351197 TI - Resonant tunneling in nanocolumns improved by quantum collimation. AB - We report on a quantum collimation effect based on surface depletion regions in AlAs/GaAs nanocolumns with an embedded resonant tunneling structure. The considered MBE-grown nanodevices have been fabricated by means of a top-down approach that employs a reproducible lithographic definition of the vertical nanocolumns. By analyzing the scaling properties of these nanodevices, we discuss how a collimation effect due to a saddle point in the confining potential can explain an improved device performance of the ultimately scaled structures at room temperature. PMID- 16351198 TI - Atomic force microscopic study of aggregation of RecA-DNA nucleoprotein filaments into left-handed supercoiled bundles. AB - RecA and its complexes with double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) are responsible for homologous recombination and DNA repair. In this study, we have observed, by atomic force microscopy (AFM), two-filament left handed superhelices of RecA-dsDNA filaments that further interwind into four- or six-filament bundles, in addition to previously reported left-handed bundles of three or six filaments. Also revealed are four-filament bundles formed by further interwinding of two intrafilament superhelices of individual filaments. Pitches of superhelices of RecA-DNA filaments are similar to each other regardless the number of component filaments, and those formed on Phix174 RFII dsDNA and pNEB206A dsDNA are measured as 339.3 +/- 6.2 nm (690 counts of pitch/2) and 321.6 +/- 11.7 nm (101 counts of pitch/2), respectively, consistent with earlier measurements made by electron microscopy with a much smaller sample size. The study of these structures provides insight into the self-interactions of RecA and RecA-like proteins, which are present in all living cells, and into the general phenomenon of bundling, which is relevant to both biological and nonbiological filaments. PMID- 16351199 TI - Metal-enhanced up-conversion fluorescence: effective triplet-triplet annihilation near silver surface. AB - Up-conversion phenomena are traditionally related to two- or multiphoton processes occurring under relatively high excitation intensities. Here we present the first results of ultralow excitation intensity (in order of Wcm(-2)) continuous-wave (CW) excited up-conversion fluorescence in Kretschmann surface plasmon geometry. The active system is a blue-emitting polymer matrix blended with metalated porphyrine macrocycles. The up-conversion fluorescence is a consequence of a two-particle triplet-triplet annihilation process (TTA). PMID- 16351200 TI - Nanoscale organic light-emitting diodes. AB - This study reports the fabrication and characterization of nanoscale organic light-emitting diodes (nano-OLEDs) based on poly[2-methoxy-5-(2'-ethylhexyloxy) 1,4-phenylene vinylene] (MEH-PPV). The nano-OLEDs were fabricated by spin casting MEH-PPV into cylindrical nanoholes lithographically patterned into silicon nitride. The electroluminescence (EL) spectrum of MEH-PPV was similar to its photoluminescence spectrum, confirming radiative decay from the same excited state. Device characteristics in the form of current density and EL versus applied electric field are presented and compared with those of a large-scale OLED. PMID- 16351201 TI - Adsorbed triblock copolymers deliver reactive iron nanoparticles to the oil/water interface. AB - Reactive zero valent iron nanoparticles can degrade toxic nonaqueous phase liquids (NAPL) rapidly in contaminated groundwater to nontoxic products in situ, provided they can be delivered preferentially to the NAPL/water (oil/water) interface. This study demonstrates the ability of novel triblock copolymers to modify the nanoiron surface chemistry in a way that both promotes their colloidal stability in aqueous suspension and drives their adsorption to the oil/water interface. The ability of the copolymers to drive adsorption is demonstrated by the ability of copolymer-modified iron nanoparticles, but not the unmodified iron nanoparticles, to stabilize oil-in-water emulsions. PMID- 16351202 TI - Radiative lifetime of excitons in carbon nanotubes. AB - We calculate the radiative lifetime and energy bandstructure of excitons in semiconducting carbon nanotubes within a tight-binding approach including the electron-hole correlations via the Bethe-Salpeter equation. In the limit of rapid interband thermalization, the radiative decay rate is maximized at intermediate temperatures and decreases at low temperature because the lowest-energy excitons are optically forbidden. The intrinsic phonons cannot scatter excitons between optically active and forbidden bands, so sample-dependent extrinsic effects that break the symmetries can play a central role. We calculate the diameter-dependent energy splittings between singlet and triplet excitons of different symmetries and the resulting dependence of radiative lifetime on temperature and tube diameter. PMID- 16351203 TI - Diameter and metallicity dependent redox influences on the separation of single wall carbon nanotubes. AB - Recently, it has become possible to separate and/or enrich fractions of single wall carbon nanotubes (SWNTs) according to type (or otherwise termed "metallicity") and diameter (d(t)). Exposure of acid-treated SWNTs to amines has shown such separation. In this contribution, we describe the underlying mechanism for this separation and provide a better description of the physicochemical properties of charge-stabilized SWNT dispersions in polar aprotic media, such as N,N-dimethylformide (DMF). With the establishment of the reversible nature of the redox chemistry, SWNT(n+) + (n/2)H2O <==> SWNT + nH(+) + (n/4)O2, amine-induced pH changes as well as variations in H2O and O2 concentration in DMF are shown to cause differential partial-reduction trends according to d(t) and metallicity. At a pH of 10, the (n,m)-SWNTs that resist complete reduction to their undoped state remain in suspension while the rest that lose their charges populate the precipitate. These d(t)- and metallicity-dependent redox and separation trends are modeled based on the Gibbs free energy and charge loss as it pertains to the (n,m)-dependent SWNT integrated density of states (I(DOS)) across the corresponding pH-induced redox jump. At a given redox potential, the relative placement of the van Hove singularities and continuum determines the amount of charge left on various (n,m)-SWNTs that governs their relative dispersion stability in DMF. PMID- 16351204 TI - Growth of Y-shaped nanorods through physical vapor deposition. AB - This work presents a proposed mechanism for fabricating Y-shaped nanorods, demonstrates the feasibility of the proposal through classical molecular dynamics simulations, and validates the simulations through magnetron sputter deposition experiments. The proposed mechanism relies primarily on the formation of stacking faults during deposition and to a lesser degree on diffusion kinetics and geometrical shadowing. Applications of the proposed mechanism may enable the design of nanorod arrays with controlled branching. PMID- 16351205 TI - Flow control through polymer-grafted smart nanofluidic channels: molecular dynamics simulations. AB - Presented are results of molecular dynamics simulations that demonstrate flow gating through a polymer-grafted nanopore as a function of effective solvent quality. Analysis of density and flow profiles from the simulations show that the difference in drag force exerted on the flowing solvent due to different polymer brush configurations produces the effective fluid gating. Shear-induced permeability changes through these nanopores has also been investigated. These results establish a critical starting point in nanofluidics from which continuum modeling can be developed to design this emerging class of smart nanoporous materials with tailor-made properties. PMID- 16351206 TI - Quantum computing via defect states in two-dimensional antidot lattices. AB - We propose a new structure suitable for quantum computing in a solid-state environment: designed defect states in antidot lattices superimposed on a two dimensional electron gas at a semiconductor heterostructure. State manipulation can be obtained with gate control. Model calculations indicate that it is feasible to fabricate structures whose energy level structure is robust against thermal dephasing. PMID- 16351207 TI - Enhanced channel modulation in dual-gated silicon nanowire transistors. AB - Dual-gated silicon nanowire (SiNW) field-effect transistors (FETs) have been fabricated by using electron-beam lithography. SiNW devices (W approximately 60 nm) exhibit an on/off current ratio greater than 10(6), which is more than 3 orders of magnitude higher than that of control devices prepared simultaneously having a large channel width (approximately 5 microm). In addition, by changing the local energy-band profile of the SiNW channel, the top gate is found to suppress ambipolar conduction effectively, which is one of the factors limiting the use of nanotube or nanowire FETs for complimentary logic applications. Two dimensional numerical simulations show that the gate-induced electrostatic control is improved as the channel width of the FETs decreases. Therefore, enhanced channel modulations can be achieved in these dual-gated SiNW devices. PMID- 16351208 TI - Ordered arrays of silicon nanowires produced by nanosphere lithography and molecular beam epitaxy. AB - Because of their importance in fundamental research and possible applications in nanotechnology and nanoelectronics, semiconductor nanowires have attracted much interest. In addition to the growth itself, the control of the size and location is an essential problem. Here we show the growth of ordered arrays of vertically aligned silicon nanowires by molecular beam epitaxy using prepatterned arrays of gold droplets on Si(111) substrates. The ordered arrays of gold particles were produced by nanosphere lithography. PMID- 16351209 TI - Low temperature growth of boron nitride nanotubes on substrates. AB - High growth temperatures (>1100 degrees C), low production yield, and impurities have prevented research progress and applications of boron nitride nanotubes (BNNTs) in the past 10 years. Here, we show that BNNTs can be grown on substrates at 600 degrees C. These BNNTs are constructed of high-order tubular structures and can be used without purification. Tunneling spectroscopy indicates that their band gap ranges from 4.4 to 4.9 eV. PMID- 16351210 TI - Additive soft-lithographic patterning of submicrometer- and nanometer-scale large area resists on electronic materials. AB - We describe a novel soft-lithographic technique possessing broad utility for the fabrication of large area, nanoscale ( approximately 100 nm) multilayer resist structures on electronic material substrates. This additive patterning method transfers ultrathin poly(dimethylsiloxane) (PDMS) decals to an underlying SiO(2) capped organic planarazation layer. The PDMS patterns serve as a latent image through which high-quality multilayer resist structures can be developed using reactive ion-beam etching. PMID- 16351211 TI - Artificial organelle: ATP synthesis from cellular mimetic polymersomes. AB - A complex cellular process was reconstructed using a multiprotein polymersome system. ATP has been produced by coupled reactions between bacteriorhodopsin, a light-driven transmembrane proton pump, and F(0)F(1)-ATP synthase motor protein, reconstituted in polymersomes. This indicates that ATP synthase maintained its ATP synthesis and therefore its motor activity in the artificial membranes. This hybrid proteopolymersome will have wide application in a number of fields ranging from the in vitro investigation of cellular metabolism to the synthesis of functional "smart" materials. PMID- 16351212 TI - High-efficiency dye-sensitized solar cell based on a nitrogen-doped nanostructured titania electrode. AB - A highly efficient dye-sensitized solar cell (DSC) was fabricated using a nanocrystalline nitrogen-doped titania electrode. The properties of the nitrogen doped titania powder, film, and solar cell were investigated. The substitution of oxygen sites with nitrogen atoms in the titania structure was confirmed by X-ray photoemission spectroscopy (XPS). The UV-vis spectrum of the nitrogen-doped powder and film showed a visible light absorption in the wavelength range from 400 to 535 nm. An enhancement of the incident photon-to-current conversion efficiency (IPCE) in the range of 380-520 nm and 550-750 nm was observed. An 8% overall conversion efficiency has been achieved. The results of the stability test indicated that the solar cell fabricated by the nitrogen-doped titania exhibited great stability. PMID- 16351213 TI - Self-assembly of size- and position-controlled ultralong nanodot chains using near-field optical desorption. AB - We report the self-assembly of a size- and position-controlled ultralong nanodot chain using a novel effect of near-field optical desorption. A sub-100-nm dot chain with a deviation of 5 nm forms at a size based on plasmon resonance, depending on the photon energy; the resulting structure forms a high-transmission efficiency nanoscale waveguide. Using this method with simple lithographically patterned substrates allows one to increase the throughput of the production of nanoscale structures dramatically at all scales. PMID- 16351214 TI - Polymeric nanoparticle preparation that eradicates tumors. AB - We report the production of poly(lactic-co-glycolic acid) nanoparticles that encapsulate the photosensitizer meso-tetraphenylporpholactol. These nanoparticles are stable and nonphototoxic upon systemic administration. Upon cellular internalization, the photosensitizer is released from the nanoparticle and becomes highly phototoxic. Irradiation with visible light results in cell specific killing of several cancer cell lines. Importantly, in vivo experiments show complete eradication of cancers in mouse models. The concept of photosensitizers with selective phototoxicity should have widespread applications in cancer therapy. PMID- 16351215 TI - Fabrication of metallic nanodots in large-area arrays by mold-to-mold cross imprinting (MTMCI). AB - We have developed a mold-to-mold cross imprint (MTMCI) process, which redefines an imprint mold with another imprint mold. By performing MTMCI on two identical imprint molds with silicon spacer nanowires in a perpendicular arrangement, we fabricated a large array of sub-30-nm silicon nanopillars. Large-area arrays of Pt dots are then produced using nanoimprint lithography with the silicon nanopillar mold. PMID- 16351216 TI - Nanopantography: a new method for massively parallel nanopatterning over large areas. AB - We report a radically different approach to the versatile fabrication of nanometer-scale preselected patterns over large areas. Standard lithography, thin film deposition, and etching are used to fabricate arrays of ion-focusing microlenses (e.g., small round holes through a metal/insulator structure) on a substrate such as a silicon wafer. The substrate is then placed in a vacuum chamber, a broad-area collimated beam of ions is directed at the substrate, and electric potentials are applied to the lens arrays such that the ions focus at the bottoms of the holes (e.g., on the wafer surface). When the wafer is tilted off normal (with respect to the ion beam axis), the focal points in each hole are laterally displaced, allowing the focused beamlets to be rastered across the hole bottoms. In this "nanopantography" process, the desired pattern is replicated simultaneously in many closely spaced holes over an area limited only by the size of the broad-area ion beam. With the proper choice of ions and downstream gaseous ambient, the method can be used to deposit or etch materials. Data show that simultaneous impingement of an Ar(+) beam and a Cl(2) effusive beam on an array of 950-nm-diam lenses can be used to etch 10-nm-diam features into a Si substrate, a reduction of 95x. Simulations indicate that the focused "beamlet" diameters scale directly with lens diameter, thus a minimum feature size of approximately 1 nm should be possible with 90-nm-diam lenses that are at the limit of current photolithography. We expect nanopantography to become a viable method for overcoming one of the main obstacles in practical nanoscale fabrication: rapid, large-scale fabrication of virtually any shape and material nanostructure. Unlike all other focused ion or electron beam writing techniques, this self-aligned method is virtually unaffected by vibrations, thermal expansion, and other alignment problems that usually plague standard nanofabrication methods. This is because the ion focusing optics are built on the wafer. PMID- 16351217 TI - Approach to steady-state transport in nanoscale conductors. AB - We show, using a tight-binding model and time-dependent density-functional theory, that a quasi-steady-state current can be established dynamically in a finite nanoscale junction without any inelastic effects. This is simply due to the geometrical constriction experienced by the electron wave packets as they propagate through the junction. We also show that in this closed nonequilibrium system two local electron occupation functions can be defined on each side of the nanojunction which approach Fermi distributions with increasing number of atoms in the electrodes. The resultant conductance and current-voltage characteristics at quasi-steady state are in agreement with those calculated within the static scattering approach. PMID- 16351218 TI - Vacancy engineered ceria nanostructures for protection from radiation-induced cellular damage. AB - The ability of engineered cerium oxide nanoparticles to confer radioprotection was examined. Human normal and tumor cells were treated with nanoceria and irradiated, and cell survival was measured. Treatment of normal cells conferred almost 99% protection from radiation-induced cell death, whereas the same concentration showed almost no protection of tumor cells. For the first time, nanoceria is shown to confer radioprotection to a normal human breast line but not to a human breast tumor line, MCF-7. PMID- 16351219 TI - [60]fullerene is a powerful antioxidant in vivo with no acute or subacute toxicity. AB - In the present work, we report the effects of C(60)-pretreatments on acute carbon tetrachloride intoxication in rats, a classical model for studying free-radical mediated liver injury. Our results show that aqueous C(60) suspensions prepared without using any polar organic solvent not only have no acute or subacute toxicity in rodents but they also protect their livers in a dose-dependent manner against free-radical damage. To be sure, according to histopathological examinations and biological tests, pristine C(60) can be considered as a powerful liver-protective agent. PMID- 16351220 TI - Two computational primitives for algorithmic self-assembly: copying and counting. AB - Copying and counting are useful primitive operations for computation and construction. We have made DNA crystals that copy and crystals that count as they grow. For counting, 16 oligonucleotides assemble into four DNA Wang tiles that subsequently crystallize on a polymeric nucleating scaffold strand, arranging themselves in a binary counting pattern that could serve as a template for a molecular electronic demultiplexing circuit. Although the yield of counting crystals is low, and per-tile error rates in such crystals is roughly 10%, this work demonstrates the potential of algorithmic self-assembly to create complex nanoscale patterns of technological interest. A subset of the tiles for counting form information-bearing DNA tubes that copy bit strings from layer to layer along their length. PMID- 16351221 TI - Ultralow-temperature superplasticity in nanoceramic composites. AB - We report the successful demonstration for low-temperature and high-strain-rate superplastic forming of nanoceramic composites for the first time. Porous preforms of nanoceramic composites that were partially densified at low temperatures were superplastically deformed by SPS at the record low temperatures of approximately 1000 to 1050 degrees C, which are comparable to those of Ni based superalloys. The maximum strain rate achieved is over 10(-2) s(-1), and a compressive strain over 200% can be obtained without cracking. The final products have nanosized grains with excellent optical properties. The present findings present a new strategy for nanoceramic superplasticity, demonstrating that a more practical application of nanoceramic superplasticity is not in the shaping of already-dense materials but in the near-net-shape forming of partially dense parts. PMID- 16351222 TI - Nanoexplosion synthesis of multimetal oxide ceramic nanopowders. AB - Herein we demonstrate a unique processing technique for engineering multicomponent ceramic nanopowders with precise morphologies by "nanoblast" calcination/deagglomeration. Multiple "nanoexplosions" of C(3)H(6)N(6)O(6) nanoparticles embedded in preliminary engineered nanoreactors break apart the agglomerates because of the highly energetic impacts of the blast waves. Also, the solid-solubility of one component into the other is enhanced by the extremely high local temperature generated during the nanoexplosions. We applied this technique to produce nanosized agglomerate-free ceria-gadolinia solid solution powder with an average aggregate size of 42 nm. The described method opens the door to the synthesis of a wide range of multimetal oxide ceramic and metal ceramic composite nanopowders, with precise stoichiometries and uniform morphologies. PMID- 16351223 TI - Interaction of polyelectrolytes and their composites with living cells. AB - Since the layer-wise polyelectrolyte deposition offers the opportunity to modify surfaces for biomedical applications, interactions and toxicity between polyelectrolytes and living cells become interesting. The aim of the present work is to determine the different factors such as contact area, charge, and transplantation site that influence the cell reaction to a specific polymer. We found that toxicity is influenced by all these factors and cannot be tested easily in a model. PMID- 16351224 TI - Spectroscopy and microscopy of spin-sensitive rectification current induced by microwave radiation. AB - Amplitude-modulated microwave radiation was applied to the junction of a scanning tunneling microscope (STM). Rectification in the tunneling current was induced by the microwave because of the nonlinearity of the junction. It is shown that this rectification current is proportional to the d(2)I/dV(2) signal and exhibits spin sensitivity. The magnetic domain at the apex of an iron-coated tip was manipulated by applying an external magnetic field. The moment of spin flip and the magnetic hysteresis were measured by changes in the spin-dependent current through the Cr(001) surface. PMID- 16351225 TI - Photoluminescence mapping of "as-grown" single-walled carbon nanotubes: a comparison with micelle-encapsulated nanotube solutions. AB - Band gap photoluminescence (PL) is observed from "as-grown" single-walled carbon nanotubes (SWNTs) in solid form. The relative PL intensities for six specific semiconducting SWNTs are compared directly to those of the micelle-encapsulated SWNTs' solutions to investigate the influence of the micelle dispersion process on PL measurements. The results indicate that sodium dodecyl sulfate (SDS) and sodium cholate (SC) selectively solubilize smaller-diameter nanotubes, whereas sodium dodecylbenzene sulfonate (SDBS) solution does not exhibit significant diameter selectivity within the diameter range studied here (d(t) = 0.829-0.966 nm). PMID- 16351226 TI - Fabrication of polystyrene latex nanostructures by nanomanipulation and thermal processing. AB - The capability to fabricate nanoscale structures is a fundamental step toward realizing the promise and potential of nanotechnology. We report on precise manipulation and thermal processing using 100-nm polystyrene latex nanoparticles. This approach is illustrated by fabricating a three-dimensional nanostructure by using an AFM tip to position nanoparticles and then thermally processing to "sinter" the particles to form a contiguous, stable structure. We suggest that this is a general approach, but the use of polystyrene latex particles offers an advantage of low-temperature processing. Use of polystyrene latex also extends the range of materials for which we have demonstrated manipulation and suggests applications including fluorescent doping and electrically conducting polymers. PMID- 16351227 TI - Microtubule gliding and cross-linked microtubule networks on micropillar interfaces. AB - We combined biochemical and topographical patterning to achieve motor-driven microtubule gliding on top of microfabricated pillar arrays with limited and controllable surface interactions of gliding microtubules. Kinesins immobilized on pillar heads pushed microtubules from the top of one micropillar to the next bridging up to 20 mum deep gaps filled with buffer solution. Distances of more than 10 mum were by-passed, and microtubule buckling was occasionally observed. The velocity distributions of microtubules gliding on poly(dimethylsiloxane) (PDMS) pillars, on flat PDMS, and on glass were found to be different, most likely due to topological and/or chemical differences between the substrates. We also used pillar arrays to suspend cross-linked microtubule networks, whose structural characteristics were governed by the topographical characteristics of the pillar pattern. These experiments open new possibilities to study the dynamics and the self-organization of motor/microtubule networks in defined topologically structured environments. PMID- 16351228 TI - Optical spectroscopy on individual amphi-PIC J-aggregates. PMID- 16351229 TI - Aligned carbon nanotube stacks by water-assisted selective etching. AB - Well-aligned, high-purity carbon nanotube (CNT) stacks of up to 10 layers fabricated in one batch process have been formed by water-assisted selective etching of carbon atoms. Etching takes place at the CNT caps as well as at the interface between CNTs and metal catalyst particles. This simple process generates high-purity CNTs and opens the CNT ends by removing the nanotube caps. High-resolution transmission electron microscopy indicates that the process does not damage CNT wall structures. A mechanism for stacked growth of CNT layers is proposed. PMID- 16351232 TI - Interplay between the ionic and electronic density profiles in liquid metal surfaces. AB - First-principles molecular-dynamics simulations have been performed for the liquid-vapor interfaces of liquid Li, Mg, Al, and Si. We analyze the oscillatory ionic and valence electronic density profiles obtained, their wavelengths, and the mechanisms behind their relative phase shift. PMID- 16351230 TI - Template-assisted growth of nominally cubic (100)-oriented three-dimensional crack-free photonic crystals. AB - Three-dimensional (3D) photonic crystals (PhCs) are now beginning to acquire functionality via the use of dopants and heterostructures. However, the self organized fabrication of large-area single crystals that are free of cracks and stacking faults has remained a challenge. We demonstrate a technology for the fabrication of (100)-oriented thin film 3D opal PhCs that exhibit no cracks over areas having no intrinsic size limit via a modified template-assisted colloidal self-assembly approach onto a patterned substrate. This technology potentially makes available large area regions of single photonic crystal, which can be used for optoelectronic devices. PMID- 16351233 TI - Coverage and nearest-neighbor dependence of adsorbate diffusion. AB - We present data on the coverage and nearest-neighbor dependences of the diffusion of CO on Cu(111) by time-lapsed scanning tunneling microscope (STM) imaging. Most notable is a maximum in diffusivity of CO at a local coverage of one molecule per 20 substrate atoms and a repulsion between CO molecules upon approach closer than three adsites, which in combination with a less pronounced increase in potential energy at the diffusion transition state, leads to rapid diffusion of CO molecules around one another. We propose a new method of evaluating STM-based diffusion data that provides all parameters necessary for the modeling of the dynamics of an adsorbate population. PMID- 16351234 TI - On the connectivity of seams of conical intersection: seam curvature. AB - The seam of conical intersection of two electronic states is said to be curved when the span of the basis vectors describing the branching plane varies along the seam. In this work degenerate perturbation theory is used to determine an approximately diabatic Hamiltonian that can reliably reproduce the potential energy surfaces in the vicinity of a point of conical intersection. This Hamiltonian provides a rigorous description of seam curvature, and a means for obtaining the full (N(int)-2)-dimensional seam of conical intersection connected to a point of conical intersection. PMID- 16351235 TI - Stochastic transition states: reaction geometry amidst noise. AB - Classical transition state theory (TST) is the cornerstone of reaction-rate theory. It postulates a partition of phase space into reactant and product regions, which are separated by a dividing surface that reactive trajectories must cross. In order not to overestimate the reaction rate, the dynamics must be free of recrossings of the dividing surface. This no-recrossing rule is difficult (and sometimes impossible) to enforce, however, when a chemical reaction takes place in a fluctuating environment such as a liquid. High-accuracy approximations to the rate are well known when the solvent forces are treated using stochastic representations, though again, exact no-recrossing surfaces have not been available. To generalize the exact limit of TST to reactive systems driven by noise, we introduce a time-dependent dividing surface that is stochastically moving in phase space, such that it is crossed once and only once by each transition path. PMID- 16351236 TI - The electrostatic potential generated by topological atoms. II. Inverse multipole moments. AB - Quantum chemical topology defines finite atoms, whose bounded electron density generates a well-defined electrostatic potential. A multipole expansion based on spherical tensors provides a potential that is formally convergent outside the divergence sphere. Part I of this series [P. L. A. Popelier and M. Rafat, Chem. Phys. Lett.376, 148 (2003)] showed that a continuous multipole expansion expands the convergence region, thereby allowing the electrostatic potential to be evaluated at short range. Here, we propose a different method, based on "inverse" multipole moments, enabling an expansion that converges everywhere. These moments are defined by inverse (i.e., negative) powers of the magnitude of the position vector describing the electron density inside the atom. We illustrate this technique on nitrogen in N(2), oxygen in H(2)O, and oxygen in the phenolic group of the amino acid tyrosine. The proposed method constitutes a considerable advance over the method presented in Part I. PMID- 16351237 TI - Gauge-origin-independent magnetizabilities of solvated molecules using the polarizable continuum model. AB - We present an implementation of the polarizable continuum model in its integral equation formulation for the calculation of the magnetizabilities of solvated molecules. The gauge-origin independence of the calculated magnetizabilities and the fast basis set convergence are ensured through the use of London atomic orbitals. Our implementation can use Hartree-Fock and multiconfigurational self consistent-field (MCSCF) wave functions as well as density-functional theory including hybrid functionals such as B3LYP. We present the results of dielectric continuum effects on water and pyridine using MCSCF wave functions, as well as dielectric medium effects on the magnetizability of the aromatic amino acids as a model for how a surrounding protein environment affects the magnetizability of these molecules. It is demonstrated that the dielectric medium effects on the magnetizability anisotropies of the aromatic amino acids may be substantial, being as large as 25% in the case of tyrosine. PMID- 16351238 TI - Efficient linear-response method circumventing the exchange-correlation kernel: theory for molecular conductance under finite bias. AB - An iterative approach for calculating the frequency domain linear response of molecular systems within time-dependent density-functional theory is presented. The method completely avoids computing the exchange-correlation kernel which is typically the most expensive step for large systems. In particular, virtual orbitals are not needed. This approach may be useful for treating the response of large systems. We give an outline of the theory and a demonstration on a jellium model of an elliptic gold cluster. A detailed theory is appended discussing the computation of conductance and ac impedance of molecular junctions under bias. PMID- 16351239 TI - A combinatorial approach to the electron correlation problem. AB - Starting from a path-integral formulation of quantum statistical mechanics expressed in a space of Slater determinants, we develop a method for the Monte Carlo evaluation of the energy of a correlated electronic system. The path integral expression for the partition function is written as a contracted sum over graphs. A graph is a set of distinct connected determinants on which paths can be represented. The weight of a graph is given by the sum over exponentially large numbers of paths which visit the vertices of the graph. We show that these weights are analytically computable using combinatorial techniques, and they turn out to be sufficiently well behaved to allow stable Monte Carlo simulations in which graphs are stochastically sampled according to a Metropolis algorithm. In the present formulation, graphs of up to four vertices have been included. In a Hartree-Fock basis, this allows for paths which include up to sixfold excitations relative to the Hartree-Fock determinant. As an illustration, we have studied the dissociation curve of the N(2) molecule in a VDZ basis, which allows comparison with full configuration-interaction calculations. PMID- 16351240 TI - Ab initio calculation of interatomic decay rates by a combination of the Fano ansatz, Green's-function methods, and the Stieltjes imaging technique. AB - A new computational technique is introduced for the ab initio calculation of the rates of interatomic and intermolecular nonradiative decay processes occurring due to electronic correlation. These recently discovered phenomena are described theoretically using the configuration-interaction formalism first introduced by Fano [Phys. Rev. 124, 1866 (1961)] and later adapted to an Auger decay by Howat et al. [J. Phys. B 11, 1575 (1978)]. The boundlike and the continuumlike components of the wave function of the decaying state are constructed using a Green's-function method known as algebraic diagrammatic construction. A combination of atomic and distributed Gaussian basis sets is shown to provide an adequate description of both boundlike and continuumlike wave-function components. The problem of the normalization of the continuum (final state) wave function is addressed using the Stieltjes imaging technique. The new method is applied to the calculation of the rates of interatomic decay in alkaline-earth rare-gas clusters. The obtained results help to verify our earlier conclusions [Phys. Rev. Lett. 93, 263002 (2004)] regarding the validity of the virtual-photon transfer model for the interatomic Coulombic decay. In addition, we demonstrate that the process of electron-transfer-mediated decay is responsible for the finite lifetimes of the outer valence vacancies in alkaline-earth-rare-gas clusters. PMID- 16351241 TI - Molecular theory of irreversibility. AB - A generalization of the Gibbs entropy postulate is proposed based on the Bogolyubov-Born-Green-Kirkwood-Yvon hierarchy of equations as the nonequilibrium entropy for a system of N interacting particles. This entropy satisfies the basic principles of thermodynamics in the sense that it reaches its maximum at equilibrium and is coherent with the second law. By using a generalization of the Liouville equation describing the evolution of the distribution vector, it is demonstrated that the entropy production is a non-negative quantity. Moreover, following the procedure of nonequilibrium thermodynamics a transport matrix is introduced and a microscopic expression for this is derived. This framework allows one to perform the thermodynamic analysis of nonequilibrium steady states with smooth phase-space distribution functions which, as proven here, constitute the states of minimum entropy production when one considers small departures from stationarity. PMID- 16351242 TI - An investigation of nodal structures and the construction of trial wave functions. AB - The factors influencing the quality of the nodal surfaces, namely, the atomic basis set, the single-particle orbitals, and the configurations included in the wave-function expansion, are examined for a few atomic and molecular systems. The following empirical rules are found: the atomic basis set must be fairly large, complete active space and natural orbitals are usually better than Hartree-Fock orbitals, multiconfiguration expansions perform better than single-determinant wave functions, but only few configurations are effective and their choice is suggested by symmetry considerations, while too long determinantal expansions spoil the nodal surfaces. These rules allow us to reduce the nodal error and to compute the best fixed node-diffusion Monte Carlo energies for a series of dimers of first-row atoms. PMID- 16351243 TI - Correlated complex independent particle potential for calculating electronic resonances. AB - We have formulated and applied an analytic continuation method for the recently formulated correlated independent particle potential [A. Beste and R. J. Bartlett J. Chem. Phys. 120, 8395 (2004)] derived from Fock space multireference coupled cluster theory. The technique developed is an advanced ab initio tool for calculating the properties of resonances in the low-energy electron-molecule collision problem. The proposed method quantitatively describes elastic electron molecule scattering below the first electronically inelastic threshold. A complex absorbing potential is utilized to define the analytic continuation for the potential. A separate treatment of electron correlation and relaxation effects for the projectile-target system and the analytic continuation using the complex absorbing potential is possible, when an approximated form of the correlated complex independent particle potential is used. The method, which is referred to as complex absorbing potential-based correlated independent particle (CAP-CIP), is tested by application to the well-known (2)Pi(g) shape resonance of e-N(2) and the (2)B(2g) shape resonance of e-C(2)H(4) (ethylene) with highly satisfactory results. PMID- 16351244 TI - The non-Markovian quantum master equation in the collective-mode representation: application to barrier crossing in the intermediate friction regime. AB - The calculation of chemical reaction rates in the condensed phase is a central preoccupation of theoretical chemistry. At low temperatures, quantum-mechanical effects can be significant and even dominant; yet quantum calculations of rate constants are extremely challenging, requiring theories and methods capable of describing quantum evolution in the presence of dissipation. In this paper we present a new approach based on the use of a non-Markovian quantum master equation (NM-QME). As opposed to other approximate quantum methods, the quantum dynamics of the system coordinate is treated exactly; hence there is no loss of accuracy at low temperatures. However, because of the perturbative nature of the NM-QME it breaks down for dimensionless frictions larger than about 0.1. We show that by augmenting the system coordinate with a collective mode of the bath, the regime of validity of the non-Markovian master equation can be extended significantly, up to dimensionless frictions of 0.5 over the entire temperature range. In the energy representation, the scaling goes as the number of levels in the relevant energy range to the third power. This scaling is not prohibitive even for chemical systems with many levels; hence we believe that the current method will find a useful place alongside the existing techniques for calculating quantum condensed-phase rate constants. PMID- 16351245 TI - Relativistic calculation of indirect NMR spin-spin couplings using the Douglas Kroll-Hess approximation. AB - We have employed the Douglas-Kroll-Hess approximation to derive the perturbative Hamiltonians involved in the calculation of NMR spin-spin couplings in molecules containing heavy elements. We have applied this two-component quasirelativistic approach using finite perturbation theory in combination with a generalized Kohn Sham code that includes the spin-orbit interaction self-consistently and works with Hartree-Fock and both pure and hybrid density functionals. We present numerical results for one-bond spin-spin couplings in the series of tetrahydrides CH(4), SiH(4), GeH(4), and SnH(4). Our two-component Hartree-Fock results are in good agreement with four-component Dirac-Hartree-Fock calculations, although a density-functional treatment better reproduces the available experimental data. PMID- 16351246 TI - Accurate and efficient treatment of two-electron contributions in quasirelativistic high-order Douglas-Kroll density-functional calculations. AB - Two-component quasirelativistic approaches are in principle capable of reproducing results from fully relativistic calculations based on the four component Dirac equation (with fixed particle number). For one-electron systems, this also holds in practice, but in many-electron systems one has to transform the two-electron interaction, which is necessary because a picture change occurs when going from the Dirac equation to a two-component method. For one-electron properties, one can take full account of picture change in a manageable way, but for the electron interaction, this would spoil the computational advantages which are the main reason to perform quasirelativistic calculations. Exploiting those picture change effects are largest in the atomic cores, which in molecular applications do not differ too much from the cores of isolated neutral atoms, we propose an elegant, efficient, and accurate approximation to the two-electron picture change problem. The new approach, called the "model potential" approach because it makes use of atomic (four- and two-component) data to estimate picture change effects in molecules, shares with the nuclear-only approach that the Douglas-Kroll operator needs to be constructed only once (not in each self consistent-field iteration) and that no time-consuming multicenter relativistic two-electron integrals need to be calculated. The new approach correctly describes the screening of both the nearest nucleus and distant nuclei, for the scalar-relativistic as well as the spin-orbit parts of the Hamiltonian. The approach is tested on atomic and molecular-orbital energies as well as spectroscopic constants of the lead dimer. PMID- 16351247 TI - Calculating initial-state-selected reaction probabilities from thermal flux eigenstates: a transition-state-based approach. AB - An approach for the calculation of initial-state-selected reaction probabilities utilizing a transition-state view and the multiconfigurational time-dependent Hartree approach is presented. Using flux correlation functions, wave packets located in the transition-state region are constructed and propagated into the asymptotic region to obtain initial-state-selected reaction probabilities. A complete set of reaction probabilities is obtained from a single set of thermal flux eigenstates. Concepts previously applied with success to the calculation of k(T) or N(E) are transferred to the calculation of state-selected probabilities. The benchmark H+H(2) (J=0) reaction on the LSTH potential-energy surface is used to test the reliability of this approach. PMID- 16351248 TI - Quantum dynamics driven by continuous laser fields under measurements: towards measurement-assisted quantum dynamics control. AB - We study quantum system dynamics driven by continuous laser fields under the measurement process. In order to take into account the system transition due to the measurement, we define the superoperator which eliminates the coherence relevant to the measured quantum states. We clarify that the dynamics of the measured states is frozen in the frequent measurement limit, while the space spanned by unmeasured states is isolated from the original system. We also derive the effective Liouvillian which governs incoherent population dynamics under the condition, in which measurements are frequently applied. We apply the formulation to two-level and Lambda-type three-level systems and clarify how the quantum measurements hinder the coherent population dynamics driven by the continuous laser fields in practical examples. Analysis on the laser field amplitude dependency of the final distribution in the t-->infinity limit suggests the possibility of the measurement-assisted quantum control. PMID- 16351249 TI - Cooperative versus dispersion effects: what is more important in an associated liquid such as water? AB - We implemented the quantum cluster equilibrium theory in our postprocessing program PEACEMAKER. This program may be run in conjunction with the very efficient vibrational frequency analysis code SNF and can therefore provide access to all electronic structure programs combined with this program. We applied the quantum cluster equilibrium theory in order to investigate the influence of a wide range of electronic structure models on the description of the liquid state. This investigation revealed much about the relevance of approximations in modern simulations of associated liquids such as water. While it is often claimed that the use of density-functional theory in condensed matter is leading to gravely erroneous results, we found that, contrary to these assertions, the exact exchange functional B3LYP and the gradient-corrected functional BP perform very well in combination with sizable basis sets as compared to second-order Moller-Plesset perturbation theory employing the same basis set. The use of density-functional theory with smaller basis sets does, in fact, lead to better results in the liquid state than the use of second-order Moller-Plesset perturbation theory in combination with these small basis sets. Most importantly, the neglect of cooperative effects disturbs a good description much more evenly if we apply second-order Moller-Plesset perturbation theory in combination with large basis sets than density-functional theory including cooperativity with smaller basis sets or Hartree-Fock using a very small basis set. PMID- 16351250 TI - Photoionization of hot radicals: C2H5, n-C3H7, and i-C3H7. AB - The combination of ion-imaging and vacuum-ultraviolet (vuv) single-photon ionization is used to study the internal energy dependence of the relative photoionization yields of the C(2)H(5),n-C(3)H(7), and i-C(3)H(7) radicals following the 266 nm photodissociation of the corresponding alkyl iodides. The comparison of the ion images obtained by vuv photoionization of the radical with those obtained by two-photon-resonant, three-photon ionization of the complementary I (2)P(32) and I*(2)P(12) atoms allows the extraction of the internal energy dependence of the cross sections. Factors influencing the appearance of the ion images in the different detection channels are discussed, including the secondary fragmentation of the neutral radicals, Franck-Condon factors for the photoionization process, and the unimolecular fragmentation of the parent photoions. PMID- 16351251 TI - An elementary method for calculating orientation-averaged fully differential electron-impact ionization cross sections for molecules. AB - Currently there are no reliable theoretical approaches for calculating fully differential cross sections (FDCS) for low-energy electron-impact ionization of large molecules. We have recently introduced the distorted-wave impulse approximation as a first step in developing improved theoretical approaches. One significant obstacle to evaluating improved theoretical approaches which require significant computer resources lies in the fact that the existing experimental data require taking averages over all molecular orientations. To circumvent this problem, it has been proposed to approximate the orientation-average by using an orientation-averaged molecular orbital in the calculation of the FDCS. The theoretical justification and expected range of validity for the approximation is given in this paper. Examples are presented for electron-impact ionization of H(2) and N(2). PMID- 16351252 TI - Does GaH5 exist? AB - The existence or nonexistence of GaH(5) has been widely discussed [N. M. Mitzel, Angew. Chem. Int. Ed. 42, 3856 (2003)]. Seven possible structures for gallium pentahydride have been systematically investigated using ab initio electronic structure theory. Structures and vibrational frequencies have been determined employing self-consistent field, coupled cluster including all single and double excitations (CCSD), and CCSD with perturbative triples levels of theory, with at least three correlation-consistent polarized-valence-(cc-pVXZ and aug-cc-pVXZ) type basis sets. The X (1)A(') state for GaH(5) is predicted to be weakly bound complex 1 between gallane and molecular hydrogen, with C(s) symmetry. The dissociation energy corresponding to GaH(5)-->GaH(3)+H(2) is predicted to be D(e)=2.05 kcal mol(-1). The H-H stretching fundamental is predicted to be v=4060 cm(-1), compared to the tentatively assigned experimental feature of Wang and Andrews [J. Phys. Chem. A 107, 11371 (2003)] at 4087 cm(-1). A second C(s) structure 2 with nearly equal energy is predicted to be a transition state, corresponding to a 90 degrees rotation of the H(2) bond. Thus the rotation of the hydrogen molecule is essentially free. However, hydrogen scrambling through the C(2v) structure 3 seems unlikely, as the activation barrier for scrambling is at least 30 kcal mol(-1) higher in energy than that for the dissociation of GaH(5) to GaH(3) and H(2). Two additional structures consisting of GaH(3) with a dihydrogen bond perpendicular to gallane (C(3v) structure 4) and an in-plane dihydrogen bond [C(s)(III) structure 5] were also examined. A C(3v) symmetry second-order saddle point has nearly the same energy as the GaH(3)+H(2) dissociation limit, while the C(s)(III) structure 5 is a transition structure to the C(3v) structure. The C(4v) structure 6 and the D(3h) structure 7 are much higher in energy than GaH(3)+H(2) by 88 and 103 kcal mol(-1), respectively. PMID- 16351253 TI - Direct-potential-fit analysis of new infrared and UV/visible A 1Sigma+-X 1Sigma+ emission spectra of AgH and AgD. AB - New high-resolution infrared and UV/visible spectra of (107)AgH, (109)AgH, (107)AgD, and (109)AgD have been recorded with a Fourier transform spectrometer. The new line positions are combined with published microwave and older electronic A (1)Sigma(+)-X (1)Sigma(+) data and used, first in a decoupled analysis of the X state alone, and then in a global multi-isotopologue analysis which yields comprehensive descriptions of both the X (1)Sigma(+) and A (1)Sigma(+) states of all four isotopologues of AgH. While the A state was long believed to be heavily perturbed, it is shown that its irregular spectrum merely reflects an unusual potential function shape. A direct fit of all data to appropriate radial Hamiltonians yields analytic potential-energy functions and Born-Oppenheimer breakdown radial functions for the ground X (1)Sigma(+) and A (1)Sigma(+) states. PMID- 16351254 TI - Electronic spectroscopy of the deuterated isotopomers of the NO.methane molecular complex. AB - The molecular complexes formed between a nitric oxide molecule and the various deuterated isotopomers of the methane molecule have been studied in a supersonic jet expansion. The electronic spectrum arising from the transition corresponding to a 3s<--pi* excitation (approximately A (2)Sigma(+)<-- approximately X (2)Pi) located on the NO chromophore has been recorded employing resonance-enhanced multiphoton ionization spectroscopy, with each of CH(4), CH(3)D, CH(2)D(2), CHD(3), and CD(4) as the complexing partner. Rich spectra are obtained, whose appearance changes in a systematic way as the amount of deuteration increases. Unexpectedly, it was possible to record spectra not only in the parent mass channel, but also in various fragment channels; this also led to the identification of some O atom resonances; and their origin is discussed. Discussion is presented of the structure in the spectra, and its possible sources including hindered internal rotation of the methane and NO moieties, overall rotation of the complex, and tunneling. In addition, some guidance has been gleaned from ab initio calculations, and these are discussed in the light of the experimental results. PMID- 16351255 TI - Cross sections and thermal rate constants for the isotope exchange reaction: D(2S)+OH(2Pi)-->OD(2Pi)+H(2S). AB - We report state-to-state and overall thermal rate constants for the isotope exchange reaction D((2)S)+OH((2)Pi)-->OD((2)Pi)+H((2)S) for 0 K750 cm(-1) excess energy) the photoelectron spectra show a loss of structure when nanosecond pulses are used, as a result of intramolecular dynamics [see Whiteside et al., J. Chem. Phys. 123, 204317 (2005), following paper]. A number of vibrational peaks in the photoelectron spectra are assigned, and we find that the common series of ion vibrational peaks observed following the ionization of p-fluorotoluene in various S(1) vibrational states is not reproduced in toluene. PMID- 16351266 TI - Photoelectron spectroscopy of S1 toluene: II. Intramolecular dynamics of selected vibrational levels in S1 toluene studied by nanosecond and picosecond time resolved photoelectron spectroscopies. AB - We present results which suggest that the photophysics of S(1) toluene is significantly more complicated than that of the related molecules p-fluorotoluene or p-difluorobenzene. We have measured a range of photoelectron spectra for a number of S(1) internal energies, on different time scales and at different temperatures, in an attempt to unravel the competing processes, but the final conclusion remains outstanding. PMID- 16351267 TI - Second OH overtone excitation and statistical dissociation dynamics of peroxynitrous acid. AB - The second OH overtone transition of the trans-perp conformer of peroxynitrous acid (tp-HOONO) is identified using infrared action spectroscopy. HOONO is produced by the recombination of photolytically generated OH and NO(2) radicals, and then cooled in a pulsed supersonic expansion. The second overtone transition is assigned to tp-HOONO based on its vibrational frequency (10 195.3 cm(-1)) and rotational band contour, which are in accord with theoretical predictions and previous observations of the first overtone transition. The transition dipole moment associated with the overtone transition is rotated considerably from the OH bond axis, as evident from its hybrid band composition, indicating substantial charge redistribution upon OH stretch excitation. The overtone band exhibits homogeneous line broadening that is attributed to intramolecular vibrational redistribution, arising from the coupling of the initially excited OH stretch to other modes that ultimately lead to dissociation. The quantum state distributions of the OH X (2)Pi (nu=0) products following first and second OH overtone excitation of tp-HOONO are found to be statistical by comparison with three commonly used statistical models. The product state distributions are principally determined by the tp-HOONO binding energy of 16.2(1) kcal mol(-1). Only a small fraction of the OH products are produced in nu=1 following the second overtone excitation, consistent with statistical predictions. PMID- 16351268 TI - Ab initio potential-energy surfaces of O2(X3Sigmag -, a1Deltag, b1Sigmag +) +O2 (X3Sigmag -, a1Deltag, b1Sigmag +): mechanism of quenching of O2 (a 1Deltag). AB - Ab initio computational studies were carried out in order to explore the possible mechanisms of quenching of O(2)(a (1)Delta(g)) by O(2)(X (3)Sigma(g) (-)): the self-quenching of O(2)(a (1)Delta(g)) and other energy-transfer processes involving two O(2) molecules. All eighteen states arising from two O(2) molecules in the X (3)Sigma(g) (-), a (1)Delta(g), and b (1)Sigma(g) (+) states are considered. After scans at the state-averaged complete active space self consistent field method to identify possible regions of crossing between states belonging to different asymptotes, complete active state second-order perturbation theory high-symmetry optimization and low-symmetry scans established that four different minima on the seams of crossing (MSXs), arising between the a+a manifold and the X+b manifold and responsible for self-quenching: O(2)(a (1)Delta(g))+O(2)(a (1)Delta(g))-->O(2)(X (3)Sigma(g) (-))+O(2)(b (1)Sigma(g) (+)), have coplanar C(2h) or C(2v) symmetries and are only 0.45 eV barrier relative to the a+a asymptote and energetically easily accessible. The rate constant for this process was estimated based on the Landau-Zener formalism. The MSXs for quenching of O(2)(a (1)Delta(g)) by the ground state O(2)(X (3)Sigma(g) (-)):O(2)(a (1)Delta(g))+O(2)(X (3)Sigma(g) (-))-->O(2)(X (3)Sigma(g) (-))+O(2)(X (3)Sigma(g) (-)) require higher energies and the process is not likely to be important. PMID- 16351269 TI - Local control of molecular fragmentation: the role of orientation. AB - Local control theory, where the instantaneous response of a system to an external field determines the control field, is employed for the purpose of inducing molecular fragmentation processes via infrared excitation. In particular, the effects of the orientational motion are investigated and compared with the idealized case of a frozen rotation. It is shown that the rotational degree of freedom is crucial for the applicability of the employed local control algorithm. The addition of an additional static electric field which induces a molecular preorientation offers an efficient way for the local control. In particular, with increasing static field strength, the fragmentation yield approaches unity so that the idealized rotationless case is recovered. Numerical results are presented for the NaI molecule. PMID- 16351270 TI - Theoretical study of Pt-Ng and Ng-Pt-Ng (Ng=Ar,Kr,Xe). AB - We have investigated the binding of noble-gas (Ng) atoms (Ng=Ar,Kr,Xe) with Pt atom by the ab initio coupled-cluster CCSD(T) method, taking into account the relativistic effects. It is shown that two Ng atoms can bind with Pt atom in linear geometry in the singlet lowest state where the second Ng atom attaches to Pt with the larger binding energy than the first Ng atom. The binding energy is evaluated as 8.2, 17.9, and 33.4 kcal/mol for Ar-Pt-Ar, Kr-Pt-Kr, and Xe-Pt-Xe, respectively, relative to the triplet ground state of the dissociation limit Pt ((3)D)+2Ng. The present results indicate that these Ng-Pt-Ng compounds are possible new gas-phase or matrix species. PMID- 16351271 TI - Stochastic dissociation of diatomic molecules. AB - The fragmentation of diatomic molecules under a stochastic force is investigated both classically and quantum mechanically, focusing on their dissociation probabilities. It is found that the quantum system is more robust than the classical one in the limit of a large number of kicks. The opposite behavior emerges for a small number of kicks. Quantum and classical dissociation probabilities do not coincide for any parameter combinations of the force. This can be attributed to a scaling property in the classical system which is broken quantum mechanically. PMID- 16351272 TI - Simulating the thermal behavior and fragmentation mechanisms of exohedral and substitutional silicon-doped C60. AB - Structures, thermal behavior, and fragmentation mechanisms of exohedral and substitutional silicon-doped C(60) containing 1-12 Si atoms are investigated by extensive molecular-dynamics simulations. A nonorthogonal tight-binding model is used to mimic the interatomic interactions in the doped fullerenes. Beginning from the minimum-energy structures, the temperature of the doped fullerenes is slowly increased until fragmentation takes place. A correlation can be established between the exohedral and substitutional structures and the corresponding fragmentation mechanisms and fragmentation temperatures. Exohedral C(60)Si(m) fullerenes fragment into two homonuclear pieces, the Si(m) cluster and the C(60) fullerene that remains intact. In contrast, the substitutional C(60 m)Si(m) heterofullerenes undergo structural transformations, including the partial unraveling of the cage, prior to fragmentation. Then, ejection of atoms or small molecules takes place from the distorted structures. The slow heating rate used, combined with long simulation runs, allows us to determine the fragmentation temperature of exohedral and substitutional Si-doped fullerenes as a function of the number of silicon atoms. Substitutional Si-doped fullerenes exhibit much higher fragmentation temperatures (1000-1500 K higher) than the exohedral fullerenes. This can be understood from the different bonding of the Si atoms in both structures. PMID- 16351273 TI - Infrared spectrum of cyclic ozone: a theoretical investigation. AB - The infrared absorption spectrum of cyclic ozone is calculated by means of a new ab initio potential energy surface, the dipole moment function, and exact quantum mechanical dynamics calculations. Five different isotopomers are considered. The absorption line for excitation of the bending fundamental near 800 cm(-1) is by far the strongest band; all other bands are more than one order of magnitude less intense. This spectral pattern as well as the isotope shifts for the various isotopomers are important for identifying cyclic ozone. Several possibilities for accessing the ring minimum of cyclic ozone are also discussed on the basis of recent electronic structure calculations. PMID- 16351274 TI - M4 @Si28 (M=Al,Ga): metal-encapsulated tetrahedral silicon fullerene. AB - It is known that silicon fullerenes cannot maintain perfect cage structures like carbon fullerenes. Previous density-functional theory calculations have shown that even with encapsulated species, nearly all endohedral silicon fullerenes exhibit highly puckered cage structures in comparison with their carbon counterparts. In this work, we present theoretical evidences that the tetrahedral fullerene cage Si(28) can be fully stabilized by encapsulating a tetrahedral metallic cluster (Al(4) or Ga(4)). To our knowledge, this is the first predicted endohedral silicon fullerene that can retain perfectly the same cage structure (without puckering) as the carbon fullerene counterpart (T(d)-C(28) fullerene). Density-functional theory calculations also suggest that the two endohedral metallosilicon fullerenes T(d)-M(4)@Si(28) (M=Al and Ga) can be chemically stable because both clusters have a large highest occupied molecular orbital-lowest unoccupied molecular orbital energy gap ( approximately 0.9 eV), strong spherical aromaticity (nucleus-independent chemical shift value of -36 and -44), and large binding and embedding energies. PMID- 16351275 TI - Optimized ensemble Monte Carlo simulations of dense Lennard-Jones fluids. AB - We apply the recently developed adaptive ensemble optimization technique to simulate dense Lennard-Jones fluids and a particle-solvent model by broad histogram Monte Carlo techniques. Equilibration of the simulated fluid is improved by sampling an optimized histogram in radial coordinates that shifts statistical weight towards the entropic barriers between the shells of the liquid. Interstitial states in the vicinity of these barriers are identified with unprecedented accuracy by sharp signatures in the quickly converging histogram and measurements of the local diffusivity. The radial distribution function and potential of mean force are calculated to high precision. PMID- 16351276 TI - 5f-->5f transitions of U4+ ions in high-field, octahedral fluoride coordination: the Cs2GeF6:U4+ crystal. AB - The U-F bond length, totally symmetric vibrational frequency, and 5f(2) energy levels of the Cs(2)GeF(6):U(4+) crystal are predicted through quantum-chemical calculations on the embedded (UF(6))(2-) cluster. The U(4+) ions substitute for much smaller Ge(4+) retaining octahedral site symmetry, which is useful to interpret the electronic transitions. The structure of the 5f(2) manifold: its energy range, the crystal splitting of the 5f(2) levels, their parentage with free-ion levels, and the energy gaps appearing within the manifold, is presented and discussed, which allows to suggest which are the possible 5f(2) luminescent levels. The effects of Cl-to-F chemical substitution are discussed by comparison with isostructural Cs(2)ZrCl(6):U(4+). The energy range of the 5f(2) manifold increases by some 6000 cm(-1) and all levels shift to higher energies, but the shift is not uniform, so that noticeable changes of order are observed from Cs(2)ZrCl(6):U(4+) to Cs(2)GeF(6):U(4+). The comparison also reveals that the green-to-blue up-conversion luminescence, which has been experimentally detected and theoretically discussed on Cs(2)ZrCl(6):U(4+), is quenched in the fluoride host. The results of the Cs(2)GeF(6):U(4+) are used as a high-symmetry model to try to understand why efficient radiative cascade emissions in the visible do not occur for charged U(4+) defects in low-symmetry YF(3) crystals. The results presented here suggest that theoretical and experimental investigations of 4f5f ions doped in octahedral, high-symmetry fluoride crystals may be conducted even when the mismatch of ionic radii between the lanthanide/actinide ions and the substituted cations of the host is considerably large. Investigations of these new materials should reveal interesting spectroscopic features without the difficulties associated with more commonly used low-symmetry fluoride hosts. PMID- 16351277 TI - High-pressure and -temperature viscosity measurements of methanol and 4:1 methanol:ethanol solution. AB - Viscosity (eta) measurements using rolling sphere viscometry in a resistance heated diamond-anvil pressure cell yield activation energies of 18-98 kJmol over a pressure range of 1.1-6.1 GPa for methanol and 26-78 kJmol over a pressure range of 2.9-5.4 GPa for a 4:1 methanol:ethanol solution. Nonlinear models of log eta (free-volume and power-law models) yield statistically better fits than the Arrhenius model at room temperature and extrapolate to lower glass-transition (10(12) Pa s) pressures. Glass-transition pressures for the free-volume model change little over the temperature range studied (298-338 K), whereas the power law values converge to those of the Arrhenius model at temperatures above 320 K. PMID- 16351278 TI - A molecular-dynamics simulation study of solvent-induced repulsion between C60 fullerenes in water. AB - Molecular-dynamics simulations of a single C(60) fullerene and pairs of C(60) fullerenes in aqueous solution have been performed for the purpose of obtaining improved understanding of the nature of solvent-induced interactions between C(60) fullerenes in water. Our simulations reveal repulsive solvent-induced interactions between two C(60) fullerenes in aqueous solution in contrast to the associative effects observed for conventional nonpolar solutes. A decomposition of the solvent-induced potential of mean force between fullerenes into entropy and energy (enthalpy) contributions reveals that the water-induced repulsion between fullerenes is energetic in origin, contrasting strongly to entropy-driven association observed for conventional nonpolar solutes. The dominance of energy in the solvent-induced interactions between C(60) fullerenes arises primarily from the high atomic density of the C(60) molecule, resulting in strong C(60) water van der Waals attraction that is reduced upon association of the fullerenes. The water-induced repulsion is found to decrease with increasing temperature due largely to an increasing contribution from a relatively weak entropy-driven association. PMID- 16351279 TI - Dynamic arrest in a liquid of symmetric dumbbells: reorientational hopping for small molecular elongations. AB - We present extensive equilibrium and out-of-equilibrium molecular-dynamics simulations of a liquid of symmetric dumbbell molecules, for constant packing fraction, as a function of temperature and molecular elongation. We compute diffusion constants as well as odd and even orientational correlators. The notations odd and even refer to the parity of the order l of the corresponding Legendre l polynomial, evaluated for the orientation of the molecular axis relative to its initial position. Rotational degrees of freedom of order l are arrested if, in the long-time limit, the corresponding orientational l correlator does not decay to zero. It is found that for large elongations translational and rotational degrees of freedom freeze at the same temperature. For small elongations only the even rotational degrees of freedom remain coupled to translational motions and arrest at a finite common temperature. On the contrary, the odd rotational degrees of freedom remain ergodic at all investigated temperatures. Hence, in the translationally arrested state, each molecule remains trapped in the cage formed by its neighboring molecules, but is able to perform 180 degrees rotations, which lead to relaxation only for the odd orientational correlators. The temperature dependence of the characteristic time of these residual rotations is well described by an Arrhenius law. Finally, we discuss the evidence in favor of the presence of the type-A transition for the odd rotational degrees of freedom, as predicted by mode-coupling theory for small molecular elongations. This transition is distinct from the type-B transition, associated with the arrest of the translational and even rotational degrees of freedom for small elongations, and with all degrees of freedom for large elongations. Odd orientational correlators are computed for small elongations at very low temperatures in the translationally arrested state. The obtained results suggest that hopping events restore the ergodicity of the odd rotational degrees of freedom at temperatures far below the A transition. PMID- 16351280 TI - Dielectric relaxation processes in water mixtures of tripropylene glycol. AB - Broadband dielectric measurements for anhydrous tripropylene glycol (3PG) and 96, 92, 84, 80, 74, 71, and 68 wt % 3PG-water mixtures are performed in the frequency range of 10(-2)-10(7) Hz and in the temperature range of 123-243 K. We examined the effect of adding water into anhydrous 3PG on relaxation dynamics. Apart from the two well-known relaxation processes, i.e., alpha and beta for anhydrous 3PG we observed new relaxation peak (beta') for all aqueous mixtures of 3PG. In addition we found the critical mole fraction of water x(w)=0.67 in which relaxation dynamics changes its behavior. According to the Sudo approach [S. Sudo et al., J. Non-Cryst. Solids 307-310, 356 (2002)], the behavior of relaxation processes was interpreted assuming the existence of three kinds of cooperative domains (CDs): containing only 3PG molecules, including only water molecules, and including both 3PG and water molecules, which molecules of each kind CD are bound by hydrogen bonds. PMID- 16351281 TI - Primary and secondary relaxations in bis-5-hydroxypentylphthalate revisited. AB - The molecular structure of bis-5-hydroxypentylphthalate (BHPP) is like dihexyl phthalate but having appended to it two hydroxyl end groups, which contribute additional dipole moments and capacity for hydrogen-bond formation. In a previously published dielectric study of the primary and secondary relaxations of BHPP, it was found that all the dynamic properties are normal except for the anomalously large width of the primary relaxation loss peak. There are two secondary relaxations, the relaxation time of the slower one increases with increasing pressure, whereas that of the faster one is practically insensitive to pressure. Hence, the slower secondary relaxation is the "universal" Johari Goldstein (JG) [J. Chem. Phys. 53, 2372 (1970); 55, 4245 (1971)] relaxation in BHPP. All is well except if the observed large width of the primary relaxation were an indication of a corresponding large coupling parameter n=0.45 in the coupling model. Then the predicted relations between the primary relaxation time tau(alpha) and the JG relaxation time tau(JG) found previously to hold in many glass formers would be violated. It was recognized that this singular behavior of BHPP is likely due to broadening of the primary loss peak by the overlapping contributions of two independent dipole moments present in BHPP, and the actual coupling parameter is smaller. However, at the time of publication of the previous work there were not enough data to support this explanation. By making broadband dielectric measurements of dibutyl phthalate (DBP) and dioctyl phthalate (DOP) that have chemical structures closely related to BHPP but with only one dipole moment, we show that all their dynamic properties are almost the same as BHPP but the widths of their primary relaxation loss peaks are significantly narrower corresponding to a smaller coupling parameter n=0.34. The new data presented here indicate that the coupling parameter of BHPP is about the same as DBP and DOP, and the predicted relations between tau(alpha) and tau(JG) of BHPP are brought back in agreement with the experimental data. PMID- 16351282 TI - Vibrational energy relaxation of the OH(D) stretch fundamental of methanol in carbon tetrachloride. AB - The lifetimes of the hydroxyl stretch fundamentals of two methanol isotopomers, MeOH and MeOD, in carbon tetrachloride solvent are calculated through the use of the perturbative Landau-Teller and fluctuating Landau-Teller methods. Examination of these systems allows for insight into the nature of the vibrational couplings that lead to intramolecular vibrational energy transfer. While both systems display energy transfer to nearly degenerate modes, MeOD also displays strong coupling to an off-resonant vibration. The relaxation of MeOH and MeOD occurs through transitions involving a total change in the vibrational quanta of 4 and 3, respectively. We calculate vibrational energy relaxation lifetimes of 4-5 ps for MeOH and 2-3 ps for MeOD that agree well with the experimentally determined values. PMID- 16351283 TI - Theory of damped quantum rotation in NMR spectra. I. Fundamental aspects. AB - The damped quantum rotation (DQR) theory, formulated originally for methyl-like atomic groupings, is now extended to hindered (N>3)-fold molecular rotors, such as the cyclopentadienyl, benzene, and cycloheptatrienyl rings in solid phase environments. It heightens the significance of the Pauli principle in shaping up the stochastic dynamics of such objects, reflected in NMR line shapes. The corresponding NMR line-shape equation is derived; its stochastic part is shown for the first time to have the double commutator form for any values of the quantum-mechanical (coherence-damping) rate constants entering it. Constraints on the relative magnitudes of such constants are determined under which the DQR line shape equation is converted into the phenomenological Alexander-Binsch equation describing classical jumps of the rotor. When all the quantum rate constants happen to be equal, the phenomenological model of equal jump rates between any two of the N (equivalent) orientations of the rotor is reproduced. On the other hand, the seemingly most plausible (for N>3) nearest-neighbor hopping model does not have any peculiar grounds in the DQR approach. For the special instances of stochastic molecular motions addressed in this work, the extended DQR formalism affords a quantification of the "degree of classicality" represented by a complete set of the relevant quantum rate constants. In view of our earlier experimental findings for the methyl rotors, the very occurrence of the nonclassical DQR effects seems unquestionable even for the objects of the size of benzene. The question of under what circumstances such effects can be big enough to be detected experimentally will be addressed in Part II of this work. PMID- 16351284 TI - Aging in a free-energy landscape model for glassy relaxation. AB - The aging properties of a simple free-energy landscape model for the primary relaxation in supercooled liquids are investigated. The intermediate scattering function and the rotational correlation functions are calculated for the generic situation of a quench from a high temperature to below the glass transition temperature. It is found that the reequilibration of molecular orientations takes longer than for translational degrees of freedom. The time scale for reequilibration is determined by that of the primary relaxation as an intrinsic property of the model. PMID- 16351285 TI - Single particle versus ensemble average: from power-law intermittency of a single quantum dot to quasistretched exponential fluorescence decay of an ensemble. AB - Light-induced diffusion-controlled electron transfer is proposed as an underlying mechanism for the intermittency (power law and breakdown) of a single quantum dot and ensemble-averaged fluorescence decay. The intensity decay can be approximated to a stretched exponential expression. The physical links to the free energy gap, reorganization energy, electronic coupling, and diffusion correlation times are discussed. A procedure is described for extracting these molecular-based parameters from experiments and is demonstrated with examples using existing data. PMID- 16351286 TI - Effective density terms in proper integral equations. AB - Two complementary routes to a new integral equation theory for site-site molecular fluids are presented. First, a simple approximation to a subset of the atomic site bridge functions in the diagrammatically proper integral equation theory is presented. This in turn leads to a form analogous to the reactive fluid theory, in which the normalization of the intramolecular distribution function and the value of the off-diagonal elements in the density matrix of the proper integral equations are the means of propagating the bridge function approximation. Second, a derivation from a topological expansion of a model for the single-site activity followed by a topological reduction and low-order truncation is given. This leads to an approximate numerical value for the new density coefficient. The resulting equations give a substantial improvement over the standard construction as shown with a series of simple diatomic model calculations. PMID- 16351287 TI - Pinning mass-selected Agn clusters on the TiO2(110)-1x1 surface via deposition at high kinetic energy. AB - We present the first scanning tunneling microscopy (STM) study of the deposition of mass-selected silver clusters (Ag(n),n=1, 2, 3) on a rutile TiO(2)(110)-1x1 surface at room temperature under hard-landing conditions. Under hard-landing conditions, only small features are observed on the surface in all cases without sintering or surface damage. This suggests that the high impact energy of the clusters mainly dissipates as thermal energy in the substrate, resulting in the recovery of any initial impact-induced surface damage and the formation of bound clusters on the surface near the impact point. STM images indicate that Ag(1) binds on the bridging oxygen rows twice as often as on the Ti rows. Density functional Theory (DFT) calculations are consistent with Ag(1) binding at either bridging oxygen vacancies or with two adjacent bridging oxygen atoms in the same bridging oxygen row. STM images of Ag(2) and Ag(3) depositions indicate almost exclusive binding centered on the Ti-atom rows. DFT calculations suggest that the Ag(2) and Ag(3) clusters are bound between two bridging oxygen rows, which is consistent with the STM observations. PMID- 16351288 TI - Manipulating the activation barrier for H2(D2) desorption from potassium-modified palladium surfaces. AB - In this work the permeation and desorption of hydrogen (deuterium) from potassium modified Pd(111) and polycrystalline palladium surfaces have been studied in the temperature range from 350 to 523 K. Time-of-flight spectroscopy has been used to determine the translational energy distributions of associatively desorbing H(2)(D(2)) molecules as a function of the potassium coverage and additional isotropic O(2) and CO background pressures. It turned out that the energy distribution of the hydrogen desorption flux is thermalized for the clean Pd surfaces but hyperthermal for the potassium-covered surfaces. The activation barrier for adsorption was found to increase with the potassium coverage but to decrease again in the presence of coadsorbates such as O(2) or CO. Especially by choosing different isotropic CO pressures, the effective desorption barrier for hydrogen could be reversibly decreased and increased, which resulted in the equivalent changes of the mean kinetic energies of the desorbing H(2) molecules. PMID- 16351289 TI - Adsorption of semiflexible block copolymers on homogeneous surfaces. AB - We present the results of extensive numerical off-lattice Monte Carlo simulations of semiflexible block-copolymer chains adsorbed onto flat homogeneous surfaces. We have compared the behavior of several chain structures, such as homopolymers, diblocks, (A(alpha)B(alpha)) block copolymers, and random heteropolymers. In all the cases studied, we have found the adsorption process to be favored with an increase of the chain rigidity. Particularly, the adsorption of diblock structures becomes a two-step process characterized by two different adsorbing temperatures that depend on the chain stiffness kappa, the chain length N, and the adsorbing energies epsilon(A) and epsilon(B). This twofold adsorbing process changes to a single one for copolymers of reduced block size alpha. Each block of the stiff copolymer chain is found to satisfy the classical scaling laws for flexible chains, however, we found the scaling exponent phi to depend on the chain stiffness. The measurement of the radius of gyration exhibits a typical behavior of a polymer chain composed of Nl(p) blobs whose persistence length follows l(p) approximately (kappa/k(B)T)(0.5) for large stiff chains. PMID- 16351290 TI - First-order mean-spherical approximation for interfacial phenomena: a unified method from bulk-phase equilibria study. AB - The recently proposed first-order mean-spherical approximation (FMSA) [Y. Tang, J. Chem. Phys. 121, 10605 (2004)] for inhomogeneous fluids is extended to the study of interfacial phenomena. Computation is performed for the Lennard-Jones fluid, in which all phase equilibria properties and direct correlation function for density-functional theory are developed consistently and systematically from FMSA. Three functional methods, including fundamental measure theory for the repulsive force, local-density approximation, and square-gradient approximation, are applied in this interfacial investigation. Comparisons with the latest computer simulation data indicate that FMSA is satisfactory in predicting surface tension, density profile, as well as relevant phase equilibria. Furthermore, this work strongly suggests that FMSA is very capable of unifying homogeneous and inhomogeneous fluids, as well as those behaviors outside and inside the critical region within one framework. PMID- 16351291 TI - Yes, pair correlations alone do determine sedimentation profiles of highly charged colloids. AB - Recent analyses of sedimentation profiles in charged colloidal suspensions are examined in view of general, somewhat underestimated, concepts. It is recalled that the standard equation for osmotic pressure equilibrium, which involves pair correlations between colloids only, remains valid in the presence of long-range Coulombic interactions. The entropy of the counterions and the electric field generated by the spatial inhomogeneity are implicitly taken into account in the colloid structure factor through the compressibility equation and local electroneutrality conditions. Limiting the macroscopic analysis to the pure electric-field contribution or, equivalently, to the ideal ionic pressure, corresponds to the incorrect, bare Debye-Huckel approximation for the colloid colloid correlations. A one-component description with reasonable Derjaguin Landau-Verwey-Overbeek-type ion-averaged effective pair potential between colloids is sufficient to derive the sedimentation profile of highly charged, weakly screened colloidal suspensions. The macroscopic electric field is directly related to the microscopic electrostatic potential around the colloids. PMID- 16351292 TI - Fast and stable recording of birefringence and holographic gratings in an azo polymethacrylate using a single nanosecond light pulse. AB - Pulsed light-induced recording in azobenzene polymers has recently been studied due to its potential use in optical storage applications. In this paper we study the photoinduced birefringence (Deltan) and holographic grating recording in an azobenzene side chain liquid-crystalline polymethacrylate irradiating with a single 4 ns light pulse at 532 nm. For some irradiation conditions, Deltan grows in less than 50 ns reaching an essentially stable value of about 10(-2). Holographic gratings have been registered using intensity and polarization patterns. Fast response and stability, similar to those of Deltan, was observed in the holographic recording process. Both light-induced anisotropy and relief contributions have been found in the case of gratings recorded using intensity patterns, relief being the dominant contribution at high recording energies. Polarization gratings have been recorded using two orthogonally circularly polarized beams. The resultant gratings showed stable efficiencies up to 0.8% (measured at 633 nm in 1-mum-thick films) and no measurable relief was observed. PMID- 16351293 TI - Large-scale molecular-dynamics simulation of nanoscale hydrophobic interaction and nanobubble formation. AB - We performed large-scale molecular-dynamics simulation of nanoscale hydrophobic interaction manifested by the formation of nanobubble between nanometer-sized hydrophobic clusters at constrained equilibrium. Particular attention is placed on the tendency of formation and stability of nanobubbles in between model nanoassemblies which are composed of hydrophobic clusters (or patches) embedded in a hydrophilic substrate. On the basis of physical behavior of nanobubble formation, we observed a change from short-range molecular hydrophobic interaction to midrange nanoscopic interaction when the length scale of hydrophobe approaches to about 1 nm. We investigated the behavior of nanobubble formation with several different patterns of nonpolar-site distribution on the nanoassemblies but always keeping a constant ratio of nonpolar to polar monomer sites. Dynamical properties of confined water molecules in between nanoassemblies are also calculated. PMID- 16351294 TI - Free-energy calculations for the cubic ZrO2 crystal as an example of a system with a soft mode. AB - We calculate the free energy for a crystalline ZrO(2) with a soft mode by the first-principles method, using the double-well energy-displacement relation. The soft-mode branch is considered as an ensemble of independent anharmonic oscillators of the parabola-plus-Gaussian or of the 2-4 polynomial forms. The anharmonic contributions are included to reproduce the cubic-to-tetragonal phase transition, however, it appears that the cubic phase does not become the most stable within the framework of the independent oscillators approach. PMID- 16351295 TI - Near-UV photodissociation of oriented CH3I adsorbed on Cu(110)-I. AB - Methyl iodide adsorbed on a Cu(110)-I surface has been found to be highly orientationally ordered. We have exploited this orientation to select different CH(3)I excited states for photodissociation by using polarized near-UV light at wavelengths of 308, 248, and 222 nm. Using p-polarized light at all three wavelengths, we find that dissociation proceeds largely via the (3)Q(0) state, consistent with the picture from gas-phase photolysis. In contrast, using s polarized light we find contributions from the (3)Q(1) state at lambda=308 nm, the (1)Q(1) state at lambda=248 nm, and the (E,1) state at lambda=222 nm-the latter being a state that has not been implicated in gas-phase studies of CH(3)I A-band photolysis. We also note the contribution to surface photodissociation from low-energy photoelectrons causing dissociative electron attachment to adsorbed CH(3)I and have identified the promotion of direct photodissociation pathways during lambda=308 nm photolysis. PMID- 16351296 TI - Adsorption dynamics of CO2 on copper-precovered ZnO(0001)-Zn: a molecular-beam scattering and thermal-desorption spectroscopy study. AB - Initial, S(0), as well as coverage-dependent adsorption probability measurements, S(Theta), have been conducted at normal impact angle and as a function of the impact energy of CO(2), E(i), adsorption temperature, T(s), and copper precoverage, Theta(Cu) (at 300 K). S(0), which decreased from approximately 0.4 exponentially to approximately 0.05 with E(i) was independent of Theta(Cu). Astonishingly, S(0) for Cu on ZnO(0001)-Zn is smaller than for the clean support which indicates a chemical modification of the support by the Cu deposits. S(Theta) curves consist of two regimes, a Kisliuk-type and Langmuirian-type section. The first is consistent with capture zone models; the second may indicate direct adsorption of CO(2) on the Cu cluster. The thermal-desorption (TDS) curves for Cu on ZnO(0001)-Zn consist of two structures with binding energies of 26 and approximately 40 kJmol (nu=1 x 10(13) ls). The TDS results indicate that CO(2) populates predominantly the Cu deposits and the rim along the Cu nanoparticles. No indications for CO(2) dissociation could be obtained with Auger electron spectroscopy. PMID- 16351297 TI - Layered interfaces between immiscible liquids studied by density-functional theory and molecular-dynamics simulations. AB - We present a study of the structure in the interface between two immiscible liquids by density-functional theory and molecular-dynamics calculations. The liquids are modeled by Lennard-Jones potentials, which achieve immiscibility by suppressing the attractive interaction between unlike particles. The density profiles of the liquids display oscillations only in a limited part of the simple liquid-phase diagram (rho,T). When approaching the liquid-vapor coexistence, a significant depletion appears while the layering behavior of the density profile vanishes. By analogy with the liquid-vapor interface and the analysis of the adsorption this behavior is suggested to be strongly related to the drying transition. PMID- 16351298 TI - Brownian dynamics of mixed surfactant micelles. AB - We investigate micelle formation in a system containing two or more different amphiphiles with different geometries using a stochastic molecular-dynamics (MD) simulation method. For a binary system containing two amphiphiles, we calculate the critical micelle concentration (CMC) and cluster distribution for the mixture at several mole fractions and compare the simulation results with those predicted by analytic theories in the dilute limit and with experiments. We find that the CMC obtained from molecular mean-field theory agrees well with our simulation results. Motivated by the industrial use of mixed surfactant systems, we then extend our studies to a system containing six different chain lengths drawn from a Poisson distribution. We find that unlike a binary mixture of amphiphiles, the different species cancel the effects of each other so that the cluster distribution for the mixture has a shape of a system consisted entirely of amphiphiles of length equal to the mean chain length of the Poisson distribution. PMID- 16351299 TI - Effect of adsorbed concentration on the radiative rate enhancement of photoexcited molecules embedded in single microspheres. AB - The variation of the molecular density in a single microcavity and its influence on the radiative rate enhancement (RRE) are reported here. The quality factors of the observed morphology-dependent resonances (MDRs) of the microcavity remain unchanged in the absence of any absorbing effects. In contrast, the MDRs tend to disappear in the presence of strong absorption even due to the self-absorption by the molecule. Time-resolved fluorescence studies reveal the fact that the value of RRE decreases with an increase in the adsorbed concentration of the molecules. The results have been explained in terms of a detuning parameter, which is a function of the refractive index of the microcavity. The increased dispersing capability of the microsphere upon increasing its molecular density has been found to be responsible for the observed decrease in RRE. PMID- 16351300 TI - Spintronics birefringence with an extended molecular loop-wire or spiral coupling. AB - A ring with spin-orbit effects coupled to a conducting wire is shown to exhibit a phase delay which is spin dependent. The key is that the coupling of the ring to the wire is over an extended spatial range and not just along a single point; this breaks the symmetry and makes the ring states couple differently to forward and backward moving wire states. This results, for properly injected spin states, in a spin-flipping probability which is dependent on the energy of the injected electron and can therefore be easily controlled. Several systems are presented and shown to exhibit this effect including the basic ring which couples to a wire as well as a ring which mediates between two wires, and a spiral between two wires. PMID- 16351301 TI - Magnetization dynamics in arrays of strongly interacting magnetic nanocrystals. AB - Arrays of 6.6 nm iron oxide nanocrystals coated with fatty acid molecules were produced using the Langmuir-Blodgett technique. The arrays had a varying number of layers stacked together, going from two dimensional to three dimensional and two different in-plane interparticle separations. While temperature-dependent ac susceptibility measurements of the isolated nanocrystals obeyed the Neel-Brown relaxation law, the array relaxation deviated significantly from this simple law. This deviation together with the observed dc field influence on the susceptibility-temperature curves, the large shifts in blocking temperatures and reduction in susceptibility-temperature curve widths on going from isolated particles to the arrays indicated collective magnetization dynamics during magnetization freezing. A scaling law analysis of this freezing dynamics yielded different powers for the two different interparticle separations with no dependence on dimensionality. In spite of the spin-glass-like behavior, it is possible that small, magnetically ordered domains of nanocrystals form at low temperature. PMID- 16351302 TI - First-principles study of some factors controlling the rate of ammonia decomposition on Ni and Pd surfaces. AB - Using the plane-wave pseudopotential method within the density-functional theory with the generalized gradient approximation for exchange and correlation potential, we have calculated adsorption energies (E(ad)), diffusion barrier, and the first dissociation barrier (E(1)) for NH(3) on Ni and Pd surfaces. While the top site is found to be preferred for NH(3) adsorption on both Ni(111) and Pd(111), its calculated diffusion barrier is substantially higher for Pd(111) than for Ni(111). We also find that during the first dissociation step (NH(3)- >NH(2)+H), NH(2) moves from the top site to the nearest hollow site on Ni(111) and Pd(111) and on the stepped surfaces, Ni(211) and Pd(211), it moves from the initial top site at the step edge to the bridge site in the same atomic chain. Meanwhile H is found to occupy the hollow sites on all four surfaces. On Ni(111), E(1) is found to be 0.23 eV higher than E(ad), while at the step of Ni(211), E(1) and E(ad) are almost equal, suggesting that the probability for the molecule to dissociate is much on the step of Ni(211). In the case of Pd(211), however, we find that the dissociation barrier is much higher than E(ad). These trends are in qualitative agreement with the experimental finding that ammonia decomposition rate is much lower on Pd than on Ni. PMID- 16351303 TI - A study of electronic structures of Pt3M (M=Ti,V,Cr,Fe,Co,Ni) polycrystalline alloys with valence-band photoemission spectroscopy. AB - The surface valence-band densities of states (DOS) of Pt(3)M (M=Ti,V,Cr,Fe,Co,Ni) polycrystalline alloys were investigated with ultraviolet photoemission spectroscopy. Upon annealing the ion-sputter-cleaned alloys at high temperatures, the observed valence-band DOS spectra clearly show the modified electronic structures on the surfaces suggesting the surface segregation of Pt as predicted in thermodynamic models. The measured d-band centers and widths for the annealed alloy surfaces show qualitatively the same trend as predicted by density functional-theory calculations based on the model of a Pt "skin" on the topmost surface layer and a subsurface layer enriched in the 3d transition metal. PMID- 16351304 TI - Optical properties of low band gap alternating copolyfluorenes for photovoltaic devices. AB - In a joint experimental and theoretical work the optical response and excited state character of two novel conjugated polymers for photovoltaic applications are studied. The polymers, alternating polyfluorene (APFO) Green 1 and APFO Green 2, are both copolymers of fluorene, thiophene, and electron accepting groups. The band gaps are extended into the red and near infrared with onsets of 780 and 1000 nm, respectively, due to alternating donor and acceptor moieties along the polymer chain. Spectroscopic ellipsometry and subsequent modeling made it possible to extract the dielectric function in the range of 260-1200 nm. Semiempirical quantum chemical calculations (ZINDO) revealed the character of the main electronic transitions in the studied spectral region. The spectral band just above 400 nm was assigned to a delocalized pi-pi* transition for both polymers. The red band lying at 622 and 767 nm in the two polymers corresponds to an electronic state mainly occupying the acceptor units and having a strong charge-transfer character. We show that the ZINDO transition energies are valuable input to the application of Lorentz oscillators in modeling of the dielectric function of the polymer material. PMID- 16351305 TI - On fitting a gold embedded atom method potential using the force matching method. AB - We fit a new gold embedded atom method (EAM) potential using an improved force matching methodology which included fitting to high-temperature solid lattice constants and liquid densities. The new potential shows a good overall improvement in agreement to the experimental lattice constants, elastic constants, stacking fault energy, radial distribution function, and fcc/hcp/bcc lattice energy differences over previous potentials by Foiles, Baskes, and Daw (FBD) [Phys. Rev. B 33, 7983 (1986)] Johnson [Phys. Rev. B 37, 3924 (1988)], and the glue model potential by Ercolessi et al. [Philos. Mag. A 50, 213 (1988)]. Surface energy was improved slightly as compared to potentials by FBD and Johnson but as a result vacancy formation energy is slightly inferior as compared to the same potentials. The results obtained here for gold suggest for other metal species that further overall improvements in potentials may still be possible within the EAM framework with an improved fitting methodology. On the other hand, we also explore the limitations of the EAM framework by attempting a brute force fit to all properties exactly which was found to be unsuccessful. The main conflict in such a brute force fit was between the surface energy and the liquid lattice constant where both could not be fitted identically. By intentionally using a very large number of spline sections for the pair potential, electron density function, and embedding energy function, we eliminated a lack of functional freedom as a possible cause of this conflict and hence can conclude that it must result from a fundamental limitation in the EAM framework. PMID- 16351306 TI - Adsorption at the liquid-vapor surface of a binary liquid mixture. AB - In a binary liquid mixture, the component possessing the lowest surface tension preferentially adsorbs at the liquid-vapor surface. In the past this adsorption behavior has been extensively investigated for critical binary liquid mixtures near the mixture's critical temperature T(c). In this fluctuation-dominated regime the adsorption is described by a universal function of the dimensionless depth zxi where xi is the bulk correlation length. Fewer studies have quantitatively examined adsorption for off-critical mixtures because, in this case, one must carefully account for both the bulk and surface crossover from the fluctuation-dominated regime (close to T(c)) to the mean-field dominated regime (far from T(c)). In this paper we compare extensive liquid-vapor ellipsometric adsorption measurements for the mixture aniline+cyclohexane at a variety of critical and noncritical compositions with the crossover theory of Kiselev and co workers [J. Chem. Phys. 112, 3370 (2000)]. PMID- 16351307 TI - Enhancement of hydrogen physisorption on graphene and carbon nanotubes by Li doping. AB - Density-functional calculations of the adsorption of molecular hydrogen on a planar graphene layer and on the external surface of a (4,4) carbon nanotube, undoped and doped with lithium, have been carried out. Hydrogen molecules are physisorbed on pure graphene and on the nanotube with binding energies about 80 90 meV/molecule. However, the binding energies increase to 160-180 meV/molecule for many adsorption configurations of the molecule near a Li atom in the doped systems. A charge-density analysis shows that the origin of the increase in binding energy is the electronic charge transfer from the Li atom to graphene and the nanotube. The results support and explain qualitatively the enhancement of the hydrogen storage capacity observed in some experiments of hydrogen adsorption on carbon nanotubes doped with alkali atoms. PMID- 16351308 TI - Mechanisms of heterogeneous crystal growth in atomic systems: insights from computer simulations. AB - In this paper we analyze the atomic-level structure of solid/liquid interfaces of Lennard-Jones fcc systems. The 001, 011, and 111 faces are examined during steady state growth and melting of these crystals. The mechanisms of crystallization and melting are explored using averaged configurations generated during these steady state runs, where subsequent tagging and labeling of particles at the interface provide many insights into the detailed atomic behavior at the freezing and melting interfaces. The interfaces are generally found to be rough and we observe the structure of freezing and melting interfaces to be very similar. Large structural fluctuations with solidlike and liquidlike characteristics are apparent in both the freezing and melting interfaces. The behavior at the interface observed under either growth or melting conditions reflects a competition between ordering and disordering processes. In addition, we observe atom hopping that imparts liquidlike characteristics to the solid side of the interfaces for all three crystal faces. Solid order is observed to extend as rough, three-dimensional protuberances through the interface, particularly for the 001 and 011 faces. We are also able to reconcile our different measures for the interfacial width and address the onset of asymmetry in the growth rates at high rates of crystal growth/melting. PMID- 16351309 TI - Local orientations of fluctuating fluid interfaces. AB - Thermal fluctuations cause the local normal vectors of fluid interfaces to deviate from the vertical direction defined by the flat mean interface position. This leads to a nonzero mean value of the corresponding polar tilt angle which renders a characterization of the thermal state of an interface. Based on the concept of an effective interface Hamiltonian we determine the variances of the local interface position and of its lateral derivatives. This leads to the probability distribution functions for the metric of the interface and for the tilt angle which allows us to calculate its mean value and its mean-square deviation. We compare the temperature dependences of these quantities as predicted by the simple capillary-wave model, by an improved phenomenological model, and by the microscopic effective interface Hamiltonian derived from density-functional theory. The mean tilt angle discriminates clearly between these theoretical approaches and emphasizes the importance of the variation of the surface tension at small wavelengths. Also the tilt angle two-point correlation function is determined which renders an additional structural characterization of interfacial fluctuations. Various experimental accesses to measure the local orientational fluctuations are discussed. PMID- 16351310 TI - ac electroosmosis in rectangular microchannels. AB - Motivated by the growing interest in ac electroosmosis as a reliable no moving parts strategy to control fluid motion in microfluidic devices for biomedical applications, such as lab-on-a-chip, we study transient and steady-state electrokinetic phenomena (electroosmosis and streaming currents) in infinitely extended rectangular charged microchannels. With the aid of Fourier series and Laplace transforms we provide a general formal solution of the problem, which is used to study the time-dependent response to sudden ac applied voltage differences in case of finite electric double layer. The Debye-Huckel approximation has been adopted to allow for an algebraic solution of the Poisson Boltzmann problem in Fourier space. We obtain the expressions of flow velocity profiles, flow rates, streaming currents, as well as expressions of the complex hydraulic and electrokinetic conductances. We analyze in detail the dependence of the electrokinetic conductance on the extension of linear dimensions relative to the Debye length, with an eye on finite electric double layer effects. PMID- 16351311 TI - Predicting internal protein dynamics from structures using coupled networks of hindered rotators. AB - Internal motions in proteins, such as oscillations of internuclear vectors u(N(i)H(i) (N)) of amide bonds about their equilibrium position, can be characterized by a local order parameter. This dynamic parameter can be determined experimentally by measuring the longitudinal and transverse relaxation rates of (15)N(i) nuclei by suitable NMR methods. In this paper, it is shown that local variations of order parameters S(ii) (2) can be predicted from the knowledge of the structure. To this effect, the diffusive motion of the internuclear vector u(N(i)H(i) (N)) is described in a potential that takes into account the deviations of the angles theta(ij) between u(N(i)H(i) (N)) and neighboring vectors u(N(j)H(j) (N)) from their average value and similarly of deviations of the angles subtended between u(N(i)H(i) (N)) and u(X(j)Y(j)), where X(j) and Y(j) are heavy atoms in the vicinity of the u(N(i)H(i) (N)) vector under investigation. It is shown how the concept of vicinity can be defined by a simple cutoff threshold, i.e., by neglecting vectors u(X(j)Y(j)) with distances d(N(i),X(j))>7.5 A. The local order parameters S(ii) (2) can be predicted from the structure using a limited set of coordinates of heavy atoms. The inclusion of a larger number of heavy atoms does not improve the predictions. Applications to calmodulin, calbindin, and interleukin 4 illustrate the success and limitations of the predictions. PMID- 16351312 TI - First-principle computation of zero-field splittings: application to a high valent Fe(IV)-oxo model of nonheme iron proteins. AB - We report the computational implementation of a combined spin-density-functional theory and perturbation theory (SDFT-PT) methodology for the accurate calculation of zero-field splittings (ZFS) in complexes of the most diverse nature including metal centers in proteins. We have applied the SDFT-PT methodology to study the cation of the recently synthesized complex [Fe(IV)(O)-(TMC)(NCCH(3))](OTf)(2), [J. Rohde et al., Science 299, 1037 (2003)] which is an important structural and functional analog of high-valent intermediates in catalytic cycles of nonheme iron enzymes. The calculated value (D(Theory)=28.67 cm(-1)) is in excellent agreement with the unusually large ZFS reported by experiment (D(Exp)=29+/-3 cm( 1)). The principal component D(zz) of the ZFS tensor is oriented along the Fe(IV)=oxo bond indicating that the oxo ligand dominates the electronic structure of the complex. PMID- 16351313 TI - Multiple time scale dynamics of distance fluctuations in a semiflexible polymer: a one-dimensional generalized Langevin equation treatment. AB - Time-dependent fluctuations in the distance x(t) between two segments along a polymer are one measure of its overall conformational dynamics. The dynamics of x(t), modeled as the coordinate of a particle moving in a one-dimensional potential well in thermal contact with a reservoir, is treated with a generalized Langevin equation whose memory kernel K(t) can be calculated from the time correlation function of distance fluctuations C(t) identical with x(0)x(t). We compute C(t) for a semiflexible continuum model of the polymer and use it to determine K(t) via the GLE. The calculations demonstrate that C(t) is well approximated by a Mittag-Leffler function and K(t) by a power-law decay on time scales of several decades. Both functions depend on a number of parameters characterizing the polymer, including chain length, degree of stiffness, and the number of intervening residues between the two segments. The calculations are compared with the recent observation of a nonexponential C(t) and a power law K(t) in the conformational dynamics within single molecule proteins [Min et al., Phys. Rev. Lett. 94, 198302 (2005)]. PMID- 16351314 TI - Bi-modal hetero-aggregation rate response to particle dosage. AB - The rate of flocculation of cationic polystyrene latex (PSL) particles by smaller, anionic PSL particles has been measured using a low-angle static light scattering technique. The rate of aggregate growth has been investigated as a function of particle size ratio and relative concentration of each particle species (for a constant dose of cationic particles). Contrary to many previous reports, two peaks in the flocculation rate were observed as a function of dose. It is speculated that the peak observed at the lower particle concentration coincides with the dose yielding maximum constant collision efficiency in the steady-state regime, a condition which is attained only after complete adsorption of the smaller particles onto the larger particle species. The peak at the higher particle concentration is believed to be related to the maximum collision rate constant upon reaching the steady-state regime, the value of which corresponds to maximum degree of aggregation and therefore the maximum mean collision efficiency prior to reaching this condition. From classical collision kinetics, the rate of aggregate growth may be represented as being proportional to the product of the collision rate constant and collision efficiency at any given time. Given then that the maximum value of these two variables coincides with different particle concentrations, the product of the response of each to particle dosage can in some cases yield a net bi-modal aggregation rate response to particle dosage. PMID- 16351315 TI - Isobaric and isochoric fragilities and the influence of volume on the temperature dependence of local segmental relaxation in polyvinylethylene networks. AB - The ratio of the isochoric and isobaric activation enthalpies at T(g) is used to quantify the relative contributions of the specific volume and temperature to the local segmental dynamics of polyvinylethylene networks. The dynamics of the networks become more dominated by temperature with increasing cross-link density, consistent with an increasing isobaric fragility. We also find a correlation between the isochoric and isobaric fragilities for the polyvinylethylene networks, in accord with a previously reported relationship between these two quantities for polymers and molecular glass-forming liquids [R. Casalini and C. M. Roland, Phys. Rev. E 72, 031503 (2005)]. PMID- 16351316 TI - Nucleation and size distribution of nucleus during induction period of polyethylene crystallization. AB - The crystallization process from supercooled melt results in the formation of nanosize nuclei in the earlier stage (induction period) through subsequent attachment or detachment of repeating unit to nuclei. The size distribution of nucleus f(N(j),t) in the induction period of nucleation process from the melts has not been experimentally confirmed yet by direct observation. The reason is that the number density of nuclei nu is too small to be detected experimentally. In our previous work, we showed the direct evidence of nucleation experimentally by means of small angle x-ray scattering (SAXS) technique. Further we have succeeded to observe the nucleation and f(N(j),t) of polymer crystallization from the melts by SAXS using synchrotron radiation. We increased nu by adding a nucleating agent to a polymer (polyethylene). The time evolution of f(N(j),t) was observed for the first time. PMID- 16351317 TI - Entropy- or enthalpy-driven collapse of strongly charged polymer chains in a one component charged fluid of counterions or coions. AB - We applied a simulation method [T. Sumi and H. Sekino, J. Chem. Phys. 122, 194910 (2005)] to an infinitely dilute polyelectrolyte immersed in one-component charged fluids in order to investigate salt effects on its collapse. In this model system, the degree of freedom of the counterion (or the coion) is considered using a density-functional theory for polymer-solvent admixtures, while the oppositely charged ions are treated as a structureless background having the opposite charge. Results obtained by these simulations show that not only the counterion but also the coion makes the polymer chain collapsed. The effects by the coion are stronger than that by the counterion. Temperature variation of the gyration radius of the polymer chain immersed in the counterion is opposite to that in the coion: while the radius of gyration decreases as the temperature decreases in the case of the counterion, it decreases as the temperature increases in the case of the coion. From these results we conclude that the former is interpreted as an enthalpy-driven collapse caused by the screening effects of the counterion, whereas the latter is interpreted as an entropy-driven one due to the translational entropy of the coion. PMID- 16351318 TI - Coalescence in semiconcentrated emulsions in simple shear flow. AB - The coalescence frequency in emulsions containing droplets with a low viscosity (viscosity ratio approximately 0.005) in simple shear flow has been investigated experimentally at several volume fractions of the dispersed phase (2%-14%) and several values of the shear rate (0.1-10 s(-1)). The evolution of the size distribution was monitored to determine the average coalescence probability from the decay of the total number of droplets. Theoretically models for two-droplet coalescence are considered, where the probability is given by P(c)=exp( tau(dr)tau(int)). Since the drainage time tau(dr) depends on the size of the two colliding droplets, and the collision time tau(int) depends on the initial orientation of the colliding droplets, the calculated coalescence probability was averaged over the initial orientation distribution and the experimental size distribution. This averaged probability was compared to the experimentally obtained coalescence frequency. The experimental results indicate that (1) to predict the average coalescence probability one has to take into account the full size distribution of the droplets; (2) the coalescence process is best described by the "partially mobile deformable interface" model or the "fully immobile deformable interface" model of Chesters [A. K. Chesters, Chem. Eng. Res. Des. 69, 259 (1991)]; and (3) independent of the models used it was concluded that the ratio tau(dr)tau(int) scales with the coalescence radius to a power (2+/-1) and with the rate of shear to a power (1.5+/-1). The critical coalescence radius R(o), above which hardly any coalescence occurs is about 10 microm. PMID- 16351319 TI - Error analysis and efficient sampling in Markovian state models for molecular dynamics. AB - In previous work, we described a Markovian state model (MSM) for analyzing molecular-dynamics trajectories, which involved grouping conformations into states and estimating the transition probabilities between states. In this paper, we analyze the errors in this model caused by finite sampling. We give different methods with various approximations to determine the precision of the reported mean first passage times. These approximations are validated on an 87 state toy Markovian system. In addition, we propose an efficient and practical sampling algorithm that uses these error calculations to build a MSM that has the same precision in mean first passage time values but requires an order of magnitude fewer samples. We also show how these methods can be scaled to large systems using sparse matrix methods. PMID- 16351320 TI - Phase diagram of complex fluids using an efficient integral equation method. AB - We present an adaptive technique for the determination of the phase diagram of fluids within the integral equation theory. It enables an efficient and accurate systematic mapping of the thermodynamic space in order to construct the binodal and spinodal lines. Results are obtained with the thermodynamically consistent integral equation proposed by Sarkisov [J. Chem. Phys. 114, 9496 (2001)] within the tangent linear technique that yields an exact differentiation of correlation functions. The generality of the numerical approach is assessed by determining both the liquid-vapor coexistence and the critical parameters of the generalized Lennard-Jones (n,6) potentials with varying repulsive part, including the hard sphere limit. PMID- 16351321 TI - On the role of collective and local molecular fluctuations in the relaxation of proton intrapair dipolar order in nematic 5CB. AB - We investigate the role that local motions and slow cooperative fluctuations have on the relaxation of the intrapair dipolar order in the nematic 5CB. With this purpose we present a theoretical and experimental systematic study which allow us to quantify the contribution from each type of molecular fluctuation to the intrapair dipolar order relaxation time, T(1D). The experimental work includes measurements of Zeeman and intrapair dipolar order relaxation times (T(1Z) and T(1D)) as a function of temperature at conventional NMR frequencies, in three complementary samples: normal and chain deuterated 4-n-pentyl-4(')-cyanobiphenyl (5CB and 5CB(d11)) and a mixture of normal 5CB and fully deuterated 4-n-pentyl-4' cyanobiphenyl (5CB(d19)), 50% in weight. Additionally we perform T(1Z) field cycling Larmor frequency-dependent measurements to obtain the spectral density of the cooperative fluctuations. The obtained results are as follows. (a) The cooperative molecular fluctuations have a strong relative weight in the relaxation of the intrapair dipolar order state, even at Larmor frequencies in the range of conventional NMR. (b) Alkyl chain rotations are an important relaxation mechanism of the intrapair dipolar order at megahertz frequencies. (c) Intermolecular fluctuations mediated by translational self-diffusion of the molecules is not an efficient mechanism of relaxation of the intrapair dipolar order. PMID- 16351322 TI - Generating inherent structures of liquids: comparison of local minimization algorithms. AB - Properties of minima obtained by two different local minimization algorithms, the conjugate gradient and the limited-memory Broyden-Fletcher-Goldfarb-Shanno, are compared for a liquid whose particles interact via a modified Lennard-Jones pair interaction. The average properties of inherent structures obtained by the two minimization procedures are shown to agree within statistical error, though for a given starting configuration the two algorithms may not quench to identical minima. PMID- 16351325 TI - 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) as a sensitizer for singlet oxygen imaging in cells: characterizing the irradiation-dependent behavior of TMPyP in a single cell. AB - Singlet molecular oxygen, a1Delta(g), can be detected from a single cell by its weak 1270 nm phosphorescence (a1Delta(g)-->X3Sigma(g)-) upon irradiation of the photosensitizer 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) incorporated into the cell. The behavior of this sensitizer in a cell, and hence the behavior of the associated singlet oxygen phosphorescence signal, depends on the conditions under which the sample is exposed to light. Upon irradiation of a neuron freshly incubated with TMPyP, the intensity of TMPyP fluorescence initially increases and there is a concomitant increase in the singlet oxygen phosphorescence intensity from the cell. These results appear to reflect a photoinduced release of TMPyP bound to DNA in the nucleus of the cell, where TMPyP tends to localize, and the subsequent relocalization of TMPyP to a different microenvironment in the cell. Upon prolonged irradiation of the cell, TMPyP photobleaches and there is a corresponding decrease in the singlet oxygen phosphorescence intensity from the cell. The data reported herein provide insight into key factors that can influence photosensitized singlet oxygen experiments performed on biological samples. PMID- 16351326 TI - The benefits of frequent positive affect: does happiness lead to success? AB - Numerous studies show that happy individuals are successful across multiple life domains, including marriage, friendship, income, work performance, and health. The authors suggest a conceptual model to account for these findings, arguing that the happiness-success link exists not only because success makes people happy, but also because positive affect engenders success. Three classes of evidence--crosssectional, longitudinal, and experimental--are documented to test their model. Relevant studies are described and their effect sizes combined meta analytically. The results reveal that happiness is associated with and precedes numerous successful outcomes, as well as behaviors paralleling success. Furthermore, the evidence suggests that positive affect--the hallmark of well being--may be the cause of many of the desirable characteristics, resources, and successes correlated with happiness. Limitations, empirical issues, and important future research questions are discussed. PMID- 16351328 TI - Nonverbal behavior and the vertical dimension of social relations: a meta analysis. AB - The vertical dimension of interpersonal relations (relating to dominance, power, and status) was examined in association with nonverbal behaviors that included facial behavior, gaze, interpersonal distance, body movement, touch, vocal behaviors, posed encoding skill, and others. Results were separately summarized for people's beliefs (perceptions) about the relation of verticality to nonverbal behavior and for actual relations between verticality and nonverbal behavior. Beliefs/perceptions were stronger and much more prevalent than were actual verticality effects. Perceived and actual relations were positively correlated across behaviors. Heterogeneity was great, suggesting that verticality is not a psychologically uniform construct in regard to nonverbal behavior. Finally, comparison of the verticality effects to those that have been documented for gender in relation to nonverbal behavior revealed only a limited degree of parallelism. PMID- 16351329 TI - Does positive affect influence health? AB - This review highlights consistent patterns in the literature associating positive affect (PA) and physical health. However, it also raises serious conceptual and methodological reservations. Evidence suggests an association of trait PA and lower morbidity and of state and trait PA and decreased symptoms and pain. Trait PA is also associated with increased longevity among older community-dwelling individuals. The literature on PA and surviving serious illness is inconsistent. Experimentally inducing intense bouts of activated state PA triggers short-term rises in physiological arousal and associated (potentially harmful) effects on immune, cardiovascular, and pulmonary function. However, arousing effects of state PA are not generally found in naturalistic ambulatory studies in which bouts of PA are typically less intense and often associated with health protective responses. A theoretical framework to guide further study is proposed. PMID- 16351327 TI - A test of major assumptions about behavior change: a comprehensive look at the effects of passive and active HIV-prevention interventions since the beginning of the epidemic. AB - This meta-analysis tested the major theoretical assumptions about behavior change by examining the outcomes and mediating mechanisms of different preventive strategies in a sample of 354 HIV-prevention interventions and 99 control groups, spanning the past 17 years. There were 2 main conclusions from this extensive review. First, the most effective interventions were those that contained attitudinal arguments, educational information, behavioral skills arguments, and behavioral skills training, whereas the least effective ones were those that attempted to induce fear of HIV. Second, the impact of the interventions and the different strategies behind them was contingent on the gender, age, ethnicity, risk group, and past condom use of the target audience in ways that illuminate the direction of future preventive efforts. PMID- 16351330 TI - Developmental psychology and public policy: progress and prospects. AB - This article outlines a framework for developmentally oriented policy research. Drawing from U. Bronfenbrenner's (1995) dynamic developmental systems theory, the authors suggest ways in which the key tenets of process, persons, context, and time can inform policy research in developmental psychology and can be used to support a causal interpretation of the results of those analyses. Conceptualizing public policies from a dynamic developmental systems perspective has a variety of implications for future research, and this article considers some of these implications. PMID- 16351331 TI - Maternal employment and child development: a fresh look using newer methods. AB - The employment rate for mothers with young children has increased dramatically over the past 25 years. Estimating the effects of maternal employment on children's development is challenged by selection bias and the missing data endemic to most policy research. To address these issues, this study uses propensity score matching and multiple imputation. The authors compare outcomes across 4 maternal employment patterns: no work in first 3 years postbirth, work only after 1st year, part-time work in 1st year, and full-time work in 1st year. Our results demonstrate small but significant negative effects of maternal employment on children's cognitive outcomes for full-time employment in the 1st year postbirth as compared with employment postponed until after the 1st year. Multiple imputation yields noticeably different estimates as compared with a complete case approach for many measures. Differences between results from propensity score approaches and regression modeling are often minimal. PMID- 16351332 TI - Maternal working conditions and child well-being in welfare-leaving families. AB - In the wake of welfare reform, thousands of low-income single mothers have transitioned into the labor market. In this article, the authors examine how the work conditions of mothers leaving welfare for employment are associated with the emotional well-being of 372 children ages 5 to 15 years. The authors examine the cumulative incidence, over a 5-year period, of maternal non-family-friendly work conditions, including long work hours, erratic work schedules, nonday shifts, and lengthy commute times, in association with children's internalizing and externalizing behavior problems and levels of positive behavior. The authors found that mothers' lengthy commute times are associated with higher levels of internalizing problem behaviors and lower levels of positive behaviors. PMID- 16351333 TI - The effect of school-based kindergarten transition policies and practices on child academic outcomes. AB - This study examined the effect of school-based kindergarten transition policies and practices on child outcomes. The authors followed 17,212 children from 992 schools in the Early Childhood Longitudinal Study-Kindergarten sample (ECLS-K) across the kindergarten school year. Hierarchical linear modeling revealed that the number of school-based transition practices in the fall of kindergarten was associated with more positive academic achievement scores at the end of kindergarten, even controlling for family socioeconomic status (SES) and other demographic factors. This effect was stronger for low- and middle-SES children than high-SES children. For low-SES children, 7 transition practices were associated with a .21 standard deviation increase in predicted achievement scores beyond 0 practices. The effect of transition practices was partially mediated by an intervening effect on parent-initiated involvement in school during the kindergarten year. The findings support education policies to target kindergarten transition efforts to increase parent involvement in low-SES families. PMID- 16351334 TI - The effects of universal pre-K on cognitive development. AB - In this study of Oklahoma's universal pre-K program, the authors relied on a strict birthday eligibility criterion to compare "young" kindergarten children who just completed pre-K to "old" pre-K children just beginning pre-K. This regression-discontinuity design reduces the threat of selection bias. Their sample consisted of 1,567 pre-K children and 1,461 kindergarten children who had just completed pre-K. The authors estimated the impact of the pre-K treatment on Woodcock-Johnson Achievement test scores. The authors found test impacts of 3.00 points (0.79 of the standard deviation for the control group) for the Letter-Word Identification score, 1.86 points (0.64 of the standard deviation of the control group) for the Spelling score, and 1.94 points (0.38 of the standard deviation of the control group) for the Applied Problems score. Hispanic, Black, White, and Native American children all benefit from the program, as do children in diverse income brackets, as measured by school lunch eligibility status. The authors conclude that Oklahoma's universal pre-K program has succeeded in enhancing the school readiness of a diverse group of children. PMID- 16351335 TI - The effectiveness of early head start for 3-year-old children and their parents: lessons for policy and programs. AB - Early Head Start, a federal program begun in 1995 for low-income pregnant women and families with infants and toddlers, was evaluated through a randomized trial of 3,001 families in 17 programs. Interviews with primary caregivers, child assessments, and observations of parent-child interactions were completed when children were 3 years old. Caregivers were diverse in race-ethnicity, language, and other characteristics. Regression-adjusted impact analyses showed that 3-year old program children performed better than did control children in cognitive and language development, displayed higher emotional engagement of the parent and sustained attention with play objects, and were lower in aggressive behavior. Compared with controls, Early Head Start parents were more emotionally supportive, provided more language and learning stimulation, read to their children more, and spanked less. The strongest and most numerous impacts were for programs that offered a mix of home-visiting and center-based services and that fully implemented the performance standards early. PMID- 16351336 TI - Impacts on children of a policy to promote employment and reduce poverty for low income parents: new hope after 5 years. AB - The impacts of New Hope, a program to increase parent employment and reduce poverty, were measured 5 years after parents were randomly assigned to program or control groups. New Hope had positive effects on children's school achievement, motivation, and social behavior, primarily for boys, across the age range 6-16. In comparison to impacts measured 2 years after program onset, effects on achievement were robust, but effects on social behavior were reduced. The program produced improvements in family income and use of organized child care and activity settings, suggesting possible pathways by which the New Hope package of policies influenced children's behavior. PMID- 16351337 TI - Child well-being in an era of welfare reform: the sensitivity of transitions in development to policy change. AB - This study examined the age-specific pattern of effects of welfare policies on child achievement. Drawing from 7 random-assignment welfare and antipoverty evaluations that provided more than 30,000 observations of children's achievement, this study found that times of developmental transition are the only periods sensitive to the changes in families brought about by these policies. More specifically, small positive effects of welfare and antipoverty policies were found for children making the transition into middle childhood, and small negative effects of these same policies were found for children making the transition out of middle childhood and into early adolescence. Effects were robust across various program groupings and could not be attributed to family characteristics that differ for children of different ages. This research informs the understanding of how changes in employment and income for low-income parents affect development across childhood. PMID- 16351338 TI - Neighborhood poverty and public policy: a 5-year follow-up of children's educational outcomes in the New York City moving to opportunity demonstration. AB - Data from the Moving to Opportunity Program, a randomized mobility experiment in which a subset of low-income minority families living in public housing in high poverty neighborhoods were given vouchers to move to low-poverty neighborhoods, were used to evaluate 1 policy approach for improving children's educational outcomes. Four hundred twenty-five New York City children were seen 2 1/2 and 5 years following relocation (mean age=14.64 years, SD=3.21 years). Analyses examining program effects on 5-year educational outcomes, accounting for 2 1/2 year outcomes, revealed that program effects on adolescent boys' achievement found at 2 1/2 years were not sustained at 5 years. Rather, male and female youths 14-20 years of age in low-poverty neighborhoods reported lower school grades and engagement relative to youths in high-poverty neighborhoods. From a policy standpoint, the complexity of enhancing low-income minority children's educational outcomes is underscored by the multiple dynamics involved--family, neighborhood, housing, and school. PMID- 16351339 TI - Effects of a family literacy program on low-literate children and their parents: findings from an evaluation of the Even Start family literacy program. AB - A randomized experiment was conducted to test the effectiveness of Even Start, a federally supported family literacy program providing early childhood education, adult education, parenting education, and joint parent-child literacy activities to children and parents from low-literate families. The evaluation of 18 Even Start projects followed 463 families for 2 years and found no statistically significant or educationally important impacts on Even Start families when they were compared with control families on child literacy outcomes, parent literacy outcomes, or parent-child interactions. The study concludes that Even Start projects were able to properly implement family literacy programs, and the observed lack of effectiveness is attributed to a combination of 2 factors: (a) a lack of full participation on the part of families and (b) instructional services that may be ineffective because of the curriculum content or the instructional approach. PMID- 16351340 TI - Perceptions of the parent-adolescent relationship: a longitudinal investigation. AB - A self-report measure of conflict and aspects of warmth in the parent-child relationship was completed by 1,330 11-year-old twins, 1,176 of whom completed the inventory again 3 years later. On average, adolescents' perceptions of the quality of the parent-child relationship declined consistently and moderately between age 11 and age 14. Conflict with parents increased, whereas all aspects of warmth decreased; changes were significantly greater for girls than boys. Variances increased with age, primarily because of increases in the magnitude of genetic effects. Heritability estimates ranged from .09 to .31 at intake and .35 to .45 at follow-up and tended to be higher for boys than girls. Changes in the parent-child relationship are interpreted as reflecting genotype-environment correlation processes whereby adolescents increasingly influence their relationships with their parents. PMID- 16351341 TI - Are genetically informed designs genetically informative? Comment on McGue, Elkins, Walden, and Iacono (2005) and quantitative behavioral genetics. AB - M. McGue, I. Elkins, B. Walden, and W. G. Iacono presented the findings from a twin study examining the relative contributions of genetic and environmental factors to the developmental trajectories of parent-adolescent relationships. From a behavioral genetics perspective, this study is well conceptualized, is well implemented, and raises some interesting developmental questions. Yet, the classic twin methodology and heritability estimates obfuscate the dynamic gene ecology transactions that underlie these social developmental trajectories. There is a growing divide between the findings of quantitative behavioral genetics, with its foundational estimate of a statistical genetic influence, and developmental molecular genetics. This comment provides a brief overview of this divide and its implications for the findings of McGue et al. as well as quantitative behavioral genetics more broadly. PMID- 16351342 TI - The limitations of behavior-genetic analyses: comment on McGue, Elkins, Walden, and Iacono (2005). AB - This article takes issue with the behavior-genetic analysis of parenting style presented by M. McGue, I. Elkins, B. Walden, and W. G. Iacono. The author argues that the attribution of their findings to inherited genetic effects was without basis because McGue et al. never indicated how those genetic effects manifested themselves. Instead, McGue et al. neglected important, and inevitable, developmental effects that most developmental psychologists understand to influence parent and adolescent behavior. The author also suggests that there is great merit in adopting the approach of developmental systems theory in understanding McGue et al.'s findings in particular and all developmental phenomena in general. PMID- 16351344 TI - The role of oral language revisited: a comment on the NICHD Early Child Care Research Network (2005). AB - This article comments on the discussion of S. A. Storch and G. J. Whitehurst's literacy development model in the article by the National Institute of Child Health and Human Development (NICHD) Early Child Care Research Network (ECCRN). Specifically, this comment focuses on concerns raised by the NICHD ECCRN that Storch and Whitehurst's model does not afford an important role to oral language in the development of early reading skill. Four important issues are presented that provide a serious and careful challenge to the model and conclusions of the NICHD ECCRN's report. PMID- 16351346 TI - The absence of a shape bias in children's word learning. AB - There is debate about whether preschool-age children interpret words as referring to kinds or to classes defined by shape similarity. The authors argue that the shape bias reported in previous studies is a task-induced artifact rather than a genuine word-learning strategy. In particular, children were forced to extend an object's novel label to one of several stand-alone, simple-shaped items, including a same-shape option from a different category and a different-shape option from the same superordinate category. Across 6 experiments, the authors found that the shape bias was eliminated (a) when the objects were more complex, (b) when they were presented in context, or (c) when children were no longer forced to choose. Moreover, children preferred the different-shape category alternatives when these were part of the same basic-level category as the target. The present experiments suggest that children seek out objects of the same kind when presented with a novel label, even if they are sometimes unable to identify the relevant kinds on their own. PMID- 16351347 TI - Early visual evoked potentials in callosal agenesis. AB - Three participants with callosal agenesis and 12 neurologically normal participants were tested on a simple reaction time task, with visual evoked potentials collected using a high-density 128-channel system. Independent components analyses were performed on the averaged visual evoked potentials to isolate the components of interest. Contrary to previous research with acallosals, evidence of ipsilateral activation was present in all 3 participants. Although ipsilateral visual components were present in all 4 unilateral conditions in the 2 related acallosal participants, in the 3rd, these were present only in the crossed visual field-hand conditions and not in the uncrossed conditions. Suggestions are made as to why these results differ from earlier findings and as to the neural mechanisms facilitating this ipsilateral activation. PMID- 16351348 TI - Neocortical disconnectivity disrupts sensory integration in Alzheimer's disease. AB - The cortical pathology in Alzheimer's disease (AD) should lead to the loss of effective interaction between distinct neocortical areas. This study compared 2 conditions within a single sensory integration task that differed in the demands placed on effective cross-cortical interaction. AD patients were impaired in their ability to bind distinct visual features of a stimulus when this binding placed greater demands on cross-cortical interaction (i.e., motion and color) but were not impaired when this binding placed lesser demands on such interaction (i.e., motion and luminance). In contrast, neurologically intact individuals and patients with Huntington's disease were able to effectively bind features under both conditions. These results provide psychophysical support for the presence of functional disconnectivity in AD and demonstrate the utility of AD for investigating the neurocognitive substrates of sensory integration. PMID- 16351349 TI - Patterns of visual scanning as predictors of emotion identification in normal aging. AB - Emotion identification appears to decline with age, and deficient visual scanning may contribute to this effect. Eye movements of 20 older adults (OAs) and 20 younger adults (YAs) with normal saccades were recorded while viewing facial expressions. OAs made fewer fixations overall, and they made a higher proportion of fixations to the lower halves of faces. Topographical distribution of fixations predicted better OA accuracy for identifying disgust than other negative emotions. Impaired OA accuracy for fear and anger was specific to vision, with normal identification of these emotions in the auditory domain. Age related frontal-lobe atrophy may affect the integrity of the frontal eye fields, with consequent scanning abnormalities that contribute to difficulties in identifying certain emotions. PMID- 16351350 TI - Repetition priming and recognition memory in younger and older persons: temporal stability and performance. AB - The dissociation between intact priming and reduced explicit memory in aging has been called into question on the basis of limited information that suggests that priming measures are less reliable than explicit memory measures. No aging study to date, however, has offered a comprehensive examination of the reliability of these measures. Age-related performance and test-retest stability coefficients for multiple priming and explicit memory tasks were compared. Age effects were found on some tasks but not others, and stability was not related to task performance. Stability was similar for implicit versus explicit task instructions, younger versus older persons, and studied versus unstudied items. Results indicate that dissociations between priming and explicit memory performances cannot be accounted for by differential reliability of the measures. PMID- 16351351 TI - Impaired implicit memory for gist information in amnesia. AB - In 2 experiments using a converging associates paradigm, the authors evaluated implicit memory for gist information in amnesic patients. In Experiment 1, participants saw multiple sets of associates, each converging on a nonpresented theme word, and were then tested using an implicit word stem completion test and an explicit cued recall test. Amnesic patients showed intact implicit and impaired explicit memory for studied words, but memory for nonpresented lures was impaired, regardless of retrieval instructions. To evaluate whether impaired implicit memory for lures was due to accelerated forgetting of gist information, short study lists were used in Experiment 2, each consisting of a single set of associates. Amnesics' implicit memory for lures was again impaired. These results point to an inability to encode robust gist representations as the cause of impaired gist memory in amnesia. PMID- 16351352 TI - Effects of age and hypertension status on cognition: the Veterans Affairs Normative Aging Study. AB - The authors examined the influence of age and hypertensive status (normotensive, controlled, untreated, or uncontrolled) on several cognitive tests via multiple regression in 357 nondemented, community-dwelling older men (mean age=67 years) whose hypertensive status was stable over 3 years and who had no medical comorbidities. Age was negatively associated with performance on all but 1 test. Age interacted with hypertensive status on verbal fluency and word list immediate recall; older uncontrolled hypertensives exhibited significantly larger age decrements on these tests compared with normotensives. These findings suggest that uncontrolled hypertension produces specific cognitive deficits beyond those attributable to age alone. These and previous findings illustrate that health conditions such as hypertension should be regularly considered in studies of "normal" cognitive aging. PMID- 16351353 TI - No, it is not possible to be schizophrenic yet neuropsychologically normal. AB - Cognitive impairment is well documented in schizophrenia, though some reports have been interpreted to suggest that it is possible to have schizophrenia without neuropsychological impairment. The authors tested this by comparing the neuropsychological profiles of closely matched patients with schizophrenia and healthy comparison participants. Sixty-four patients with schizophrenia and 64 healthy comparison cases, matched to within 3 Full-Scale IQ points, were tested using the Wechsler Adult Intelligence Scale (3rd ed.; D. Wechsler, 1997b) and the Wechsler Memory Scale (3rd ed.; D. Wechsler, 1997c). Neuropsychological profiles for these groups were markedly different, with the group of patients with schizophrenia exhibiting performance deficits in memory and speeded visual processing but superior verbal comprehension and perceptual organization relative to the group of healthy comparison participants matched on Full-Scale IQ. Thus, scoring in the normal range does not preclude neuropsychological abnormality in schizophrenia, confirming that neuropsychological impairment is a core feature of the illness. PMID- 16351354 TI - Early development of neurophysiological processes involved in normal reading and reading disability: a magnetic source imaging study. AB - This longitudinal study examined the development of the brain mechanism involved in phonological decoding in beginning readers using magnetic source imaging. Kindergarten students were assigned to 2 groups: those who showed mastery of skills that are important predictors of proficient reading (low-risk group) and those who initially did not show mastery but later benefited from systematic reading instruction and developed average-range reading skills at the end of Grade 1 (high-risk responders). Spatiotemporal profiles of brain activity were obtained during performance of letter-sound and pseudoword naming tasks before and after Grade 1 instruction. With few exceptions, low-risk children showed early development of brain activation profiles that are typical of older skilled readers. Provided that temporoparietal and visual association areas were recruited into the brain mechanism that supported reading, the majority of high risk responder children benefited from systematic reading instruction and developed adequate reading abilities. PMID- 16351356 TI - Discriminating patients with frontal-lobe epilepsy and temporal-lobe epilepsy: utility of a multilevel design fluency test. AB - Patients with frontal-lobe epilepsy (FLE) or temporal-lobe epilepsy (TLE) and matched control participants were given a design fluency test that assessed nonverbal fluency and switching ability. Patients with FLE generated fewer designs in the switching condition relative to the TLE patients and controls, whereas group differences did not emerge in the basic fluency conditions. When the side of the seizure focus and the presence or absence of a structural lesion were considered in patients with FLE, only those with left-lesional FLE generated fewer designs than controls did in the switching condition. Furthermore, patients with left-lesional and nonlesional FLE produced a greater proportion of set-loss errors than did controls. These results indicate that patients with FLE are impaired when they must simultaneously generate new designs and engage in cognitive switching; however, the pattern of impairment may depend on the side of the seizure focus and the presence of a structural lesion. PMID- 16351355 TI - Dissociations in hippocampal and frontal contributions to episodic memory performance. AB - The hippocampus and frontal lobes both contribute to episodic memory performance. In the present study, the authors evaluated the relative contributions of hippocampus, frontal lobes, anterior temporal cortex, and posterior cortex to memory performance in neurodegenerative patients and normal older controls. Subjects (n=42) were studied with structural MRI and a memory paradigm that measured delayed recall, semantic clustering during recall, recognition discriminability, and recognition response bias. Data were analyzed with multiple regression. Consistent with the authors' hypotheses, hippocampal volumes were the best predictor of delayed recall and recognition discriminability, whereas frontal volumes were the best predictor of semantic clustering and response bias. Smaller frontal volumes were associated with less semantic clustering during recall and a more liberal response bias. Results indicate that hippocampal and frontal contributions to episodic memory can be dissociated, with the hippocampus more important for memory accuracy, and frontal structures more important for strategic processing and decision making. PMID- 16351357 TI - A convergent-divergent approach to context processing, general intellectual functioning, and the genetic liability to schizophrenia. AB - Convergent and divergent validity are critically important in developing psychological measures that reveal interpretable deficits in disordered populations. This article reports on 2 studies that evaluated the validity of context processing measures. In Experiment 1, a confirmatory factor analysis of data from 481 healthy adults established the convergent validity of 2 context processing measures and showed that context processing accounted for significant amounts of variance in standard IQ and working memory measures. In Experiment 2, 20 schizophrenia patients, 16 of their healthy siblings, and 28 controls were evaluated using a novel, short context processing measure, the dot pattern expectancy (DPX) task. The DPX was sensitive to specific deficits in schizophrenia patients and their healthy siblings. These findings support the construct validity of context processing measures, suggest context processing is a component of intellectual functioning, and demonstrate that brief context processing measures remain sensitive to psychopathological deficits. PMID- 16351358 TI - Asymmetries in global-local processing ability in elderly people with the apolipoprotein e-epsilon4 allele. AB - Previous studies have identified cognitive asymmetries in elderly people at increased risk for Alzheimer's disease (AD) by comparing standardized neuropsychological tests of verbal and spatial abilities in both preclinical AD and apolipoprotein epsilon4+ elderly groups. This prospective study investigated cognitive asymmetries within a single test by comparing cognitively intact elderly (with and without the epsilon4+ allele) on a learning and memory measure that uses global and local visuospatial stimuli. Both groups demonstrated comparable overall learning and recall. But the epsilon4+ group had a significantly larger discrepancy between their global and local learning scores and had a greater proportion of individuals with more than a one standard deviation difference between their immediate recall of the global and local elements, relative to the epsilon4- group. These findings build on previous studies identifying subgroups of elderly people at greater risk for AD who often demonstrate increased cognitive asymmetries relative to groups without significant risk factors. PMID- 16351359 TI - Scaling of visuospatial attention undergoes differential longitudinal change as a function of APOE genotype prior to old age: results from the NIMH BIOCARD study. AB - The effect of apolipoprotein E (APOE) genotype on longitudinal cognitive decline in midlife was investigated with attentional scaling. Healthy individuals (mean age 59.6 years) genotyped for APOE were tested at 3 12-month intervals on a cued visual search task. A random effects model revealed significant interaction in effect of precue size on search speed between APOE-epsilon4 gene dose and assessment, with longitudinal increases in noncarriers and heterozygotes but longitudinal decreases in homozygotes. Association of APOE-epsilon4 with cognitive decline in midlife is consistent with an Alzheimer's disease (AD) prodrome, albeit a decade or more before average age of AD diagnosis. However, cognitive decline in midlife associated with a gene modulating neuronal response to insult argues that the concept of an AD prodrome includes factors that allow as well as cause AD. PMID- 16351360 TI - Priming a new identity: self-monitoring moderates the effects of nonself primes on self-judgments and behavior. AB - When a construct is primed, people often act in construct-consistent ways. Several accounts for this effect have been offered, including ideomotor theory and a social functional perspective. The authors tested an additional perspective, the Active-Self account, whereby primes can temporarily alter self perceptions. In Study 1, non-African American participants reported feeling more aggressive on an implicit measure following an African American prime. In Study 2, participants reported feeling luckier on an implicit measure following a number 7 (vs. 13) prime. In both studies, these effects were obtained only for low self-monitors, who are more likely to change self-conceptions in response to diagnostic self-information and to use their internal states in guiding behavior. Study 3 showed that low self-monitors also show larger behavioral effects of primes. PMID- 16351361 TI - Accessibility effects on implicit social cognition: the role of knowledge activation and retrieval experiences. AB - Performance on measures of implicit social cognition has been shown to vary as a function of the momentary accessibility of relevant information. The present research investigated the mechanisms underlying accessibility effects of self generated information on implicit measures. Results from 3 experiments demonstrate that measures based on response compatibility processes (e.g., Implicit Association Test, affective priming with an evaluative decision task) are influenced by subjective feelings pertaining to the ease of retrieving relevant information from memory, whereas measures based on stimulus compatibility processes (e.g., semantic priming with a lexical-decision task) are influenced by direct knowledge activation in associative memory. These results indicate that the mediating mechanisms underlying context effects on implicit measures can differ as a function of the task even when these tasks show similar effects on a superficial level. Implications for research on implicit social cognition and the ease-of-retrieval effect are discussed. PMID- 16351362 TI - Understanding others: the face and person construal. AB - The face is a critical stimulus in person perception, yet little research has considered the efficiency of the processing operations through which perceivers glean social knowledge from facial cues. Integrating ideas from work on social cognition and face processing, the current research considered the ease with which invariant aspects of person knowledge can be extracted from faces under different viewing and processing conditions. The results of 2 experiments demonstrated that participants extracted knowledge pertaining to the sex and identity of faces in both upright and inverted orientations, even when the faces were irrelevant to the task at hand. The results of an additional experiment, however, suggested that although the extraction of person knowledge from faces may occur unintentionally, the process is nonetheless contingent on the operation of a semantic processing goal. The authors consider the efficiency of person construal and the processes that support this fundamental facet of social cognitive functioning. PMID- 16351363 TI - Stereotype threat and group differences in test performance: a question of measurement invariance. AB - Studies into the effects of stereotype threat (ST) on test performance have shed new light on race and sex differences in achievement and intelligence test scores. In this article, the authors relate ST theory to the psychometric concept of measurement invariance and show that ST effects may be viewed as a source of measurement bias. As such, ST effects are detectable by means of multi-group confirmatory factor analysis. This enables research into the generalizability of ST effects to real-life or high-stakes testing. The modeling approach is described in detail and applied to 3 experiments in which the amount of ST for minorities and women was manipulated. Results indicate that ST results in measurement bias of intelligence and mathematics tests. PMID- 16351364 TI - "Where are you really from?": Asian Americans and identity denial. AB - Five studies investigate identity denial, the situation in which an individual is not recognized as a member of an important in-group. Asian Americans are seen as less American than other Americans (Study 1) and realize this is the case, although they do not report being any less American than White Americans (Studies 2A and 2B). Identity denial is a common occurrence in Asian Americans' daily lives (Study 3). They react to instances of identity denial by presenting American cultural knowledge and claiming greater participation in American practices (Studies 4 & 5). Identity denial furthers the understanding of group dynamics by capturing the experience of less prototypical group members who desire to have their common in-group identity recognized by fellow group members. PMID- 16351365 TI - The evolution and function of adult attachment: a comparative and phylogenetic analysis. AB - Although the evolutionary functions of attachment in infant-caregiver relationships are undisputed, it is unclear what functions--if any--attachment serves in adult romantic relationships. The objective of this research was to examine the evolution and function of adult attachment (i.e., pair bonding) by applying comparative and phylogenetic methods to archival data collected on 2 diverse samples of mammalian species. The authors found that species exhibiting adult attachment were more likely than others to be characterized by paternal care, developmental immaturity or neoteny, small social groups, and small body sizes. The authors also used phylogenetic techniques to reconstruct the evolution of adult attachment and test alternative evolutionary models of the comparative correlates of pair bonding. Phylogenetic analyses suggested that the relationship between paternal care and adult attachment may be a functional one (i.e., due to convergent evolution) but that the relationship between neoteny and adult attachment may be due to homology (i.e., shared ancestry). Discussion focuses on the potential of comparative and phylogenetic methods for advancing the science of social and personality psychology. PMID- 16351366 TI - Individuality and social influence in groups: inductive and deductive routes to group identity. AB - A distinction between forms of social identity formation in small interactive groups is investigated. In groups in which a common identity is available or given, norms for individual behavior may be deduced from group properties (deductive identity). In groups in which interpersonal relations are central, a group identity may also be induced from individual group members' contributions, making individuality and individual distinctiveness a defining feature of the group (inductive identity). Two studies examined the prediction that depersonalization produced by anonymity has opposite effects for groups in which social identity has been induced or deduced. Results confirmed the prediction that depersonalization increases social influence in groups whose identity was more deductive. In contrast, depersonalization decreases social influence in inductive identity groups. Implications for the role of social identity in small groups are discussed. ((c) 2005 APA, all rights reserved). PMID- 16351367 TI - Snatching defeat from the jaws of victory: self-esteem differences in the experience and anticipation of success. AB - Successes--defined broadly as meeting important standards or receiving positive evaluations--are widely assumed to be enjoyed equally by people with high self esteem (HSEs) and low self-esteem (LSEs). Three studies examined the contradictory hypothesis that HSEs react more favorably to success than do LSEs and that success brings about certain unfavorable consequences for LSEs. Undergraduate participants reacted to a laboratory-manipulated success (Studies 1 and 2) or imagined highly positive events in the future (Study 3). Self-esteem differences emerged in anxiety, thoughts about the self, and (in Study 3) thoughts about non-self-related aspects of the event. LSEs were more anxious than HSEs after succeeding, success improved HSEs' self-relevant thoughts but not LSEs', and LSEs focused more on success's negative aspects. PMID- 16351368 TI - Striving for unwanted goals: stress-dependent discrepancies between explicit and implicit achievement motives reduce subjective well-being and increase psychosomatic symptoms. AB - Three studies investigated the relevance of affect regulation, stressful life events, and congruence between explicit achievement orientation and implicit achievement motive for subjective well-being and symptom formation. According to personality systems interactions (PSI) theory, stressful life events were expected to reduce motive congruence when the ability to self-regulate affect was impaired (i.e., state orientation). Consistent with expectations, the State Orientation x Stress interaction predicted incongruence in healthy participants (Studies 1 and 3) and in patients (Study 2). Furthermore, incongruence partially mediated the direct State Orientation x Stress effect on subjective well-being (Studies 1 and 3) and the course of psychosomatic complaints over 3 months (Study 2). In Study 3, the experimental induction of threat reduced motive congruence in state-oriented participants compared with an acceptance condition. Findings underscore the importance of assessing motive congruence as a "hidden stressor" and validate a new operant multi-motive test. PMID- 16351369 TI - The structure of goal contents across 15 cultures. AB - The authors investigated the structure of goal contents in a group of 1,854 undergraduates from 15 cultures around the world. Results suggested that the 11 types of goals the authors assessed were consistently organized in a circumplex fashion across the 15 cultures. The circumplex was well described by positioning 2 primary dimensions underlying the goals: intrinsic (e.g., self-acceptance, affiliation) versus extrinsic (e.g., financial success, image) and self transcendent (e.g., spirituality) versus physical (e.g., hedonism). The circumplex model of goal contents was also quite similar in both wealthier and poorer nations, although there were some slight cross-cultural variations. The relevance of these results for several theories of motivation and personality are discussed. PMID- 16351370 TI - Attachment, caregiving, and altruism: boosting attachment security increases compassion and helping. AB - Recent studies based on J. Bowlby's (1969/1982) attachment theory reveal that both dispositional and experimentally enhanced attachment security facilitate cognitive openness and empathy, strengthen self-transcendent values, and foster tolerance of out-group members. Moreover, dispositional attachment security is associated with volunteering to help others in everyday life and to unselfish motives for volunteering. The present article reports 5 experiments, replicated in 2 countries (Israel and the United States), testing the hypothesis that increases in security (accomplished through both implicit and explicit priming techniques) foster compassion and altruistic behavior. The hypothesized effects were consistently obtained, and various alternative explanations were explored and ruled out. Dispositional attachment-related anxiety and avoidance adversely influenced compassion, personal distress, and altruistic behavior in theoretically predictable ways. As expected, attachment security provides a foundation for care-oriented feelings and caregiving behaviors, whereas various forms of insecurity suppress or interfere with compassionate caregiving. PMID- 16351371 TI - Remembering the good, forgetting the bad: intentional forgetting of emotional material in depression. AB - The authors examined intentional forgetting of negative material in depression. Participants were instructed to not think about emotional nouns that they had learned to associate with a neutral cue word. The authors provided participants with multiple occasions to suppress the unwanted words. Overall, depressed participants successfully forgot negative words. Moreover, the authors obtained a clear practice effect. However, forgetting came at a cost: Compared with the nondepressed participants and with the depressed participants who were instructed to forget positive words, depressed participants who were instructed to forget negative words showed significantly worse recall of the baseline words. These results indicate that training depressed individuals in intentional forgetting could prove to be an effective strategy to counteract automatic ruminative tendencies and mood-congruent biases. PMID- 16351372 TI - Introduction to the special section: toward a dimensionally based taxonomy of psychopathology. AB - Much current psychopathology research is framed by categorical constructs. Limitations of categorical constructs have been articulated, and dimensional constructs are often proposed as viable alternatives to categories of psychopathology. The purpose of this Special Section is to articulate and discuss diverse issues that arise in contemplating dimensional constructs as targets for psychopathology research. PMID- 16351373 TI - Diagnostic categories or dimensions? A question for the Diagnostic And Statistical Manual Of Mental Disorders--fifth edition. AB - The question of whether mental disorders are discrete clinical conditions or arbitrary distinctions along dimensions of functioning is a long-standing issue, but its importance is escalating with the growing recognition of the frustrations and limitations engendered by the categorical model. The authors provide an overview of some of the dilemmas of the categorical model, followed by a discussion of research that addresses whether mental disorders are accurately or optimally classified categorically or dimensionally. The authors' intention is to document the importance of this issue and to suggest that future editions of the Diagnostic and Statistical Manual of Mental Disorders give more recognition to dimensional models of classification. They conclude with a dimensional mental disorder classification that they suggest provides a useful model. PMID- 16351374 TI - Temperament as a unifying basis for personality and psychopathology. AB - Personality and psychopathology long have been viewed as related domains, but the precise nature of their relations remains unclear. Through most of the 20th century, they were studied as separate fields; within psychopathology, clinical syndromes were separated from personality disorders in 1980. This division led to the revelation of substantial overlap among disorders both within and across axes and to the joint study of normal and abnormal personality. The author reviews these literatures and proposes an integrative framework to explain personality psychopathology relations: Three broad, innate temperament dimensions--negative affectivity, positive affectivity, and disinhibition--differentiate through both biologically and environmentally based developmental processes into a hierarchical personality trait structure and, at their extremes, are risk factors (diatheses) for psychopathology, especially given adverse life experiences (stress). PMID- 16351375 TI - Rethinking the mood and anxiety disorders: a quantitative hierarchical model for DSM-V. AB - The fourth edition of the Diagnostic and Statistical Manual of Mental Disorders (American Psychiatric Association, 1994) groups disorders into diagnostic classes on the basis of the subjective criterion of "shared phenomenological features." There are now sufficient data to eliminate this rational system and replace it with an empirically based structure that reflects the actual similarities among disorders. The existing structural evidence establishes that the mood and anxiety disorders should be collapsed together into an overarching class of emotional disorders, which can be decomposed into 3 subclasses: the bipolar disorders (bipolar I, bipolar II, cyclothymia), the distress disorders (major depression, dysthymic disorder, generalized anxiety disorder, posttraumatic stress disorder), and the fear disorders (panic disorder, agoraphobia, social phobia, specific phobia). The optimal placement of other syndromes (e.g., obsessive-compulsive disorder) needs to be clarified in future research. PMID- 16351377 TI - Dimensional versus categorical classification of mental disorders in the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders and beyond: comment on the special section. AB - The value of including dimensional elements in the Diagnostic and Statistical Manual of Mental Disorders (DSM) has been recognized for decades. Nevertheless, no proposals have been made for introducing dimensional classification in the diagnostic system in a valid and feasible manner. As an initial step in this endeavor, the authors suggest introducing dimensional severity ratings to the extant diagnostic categories and criteria sets. Although not without difficulties, this would begin to determine the feasibility of dimensional classification and would address some limitations of the purely categorical approach (e.g., failure to capture individual differences in disorder severity, and clinically significant features subsumed by other disorders or falling below conventional DSM thresholds). The utility of incorporating broader dimensions of temperament and personality in diagnostic systems beyond the fifth edition of the DSM is also discussed. PMID- 16351376 TI - Externalizing psychopathology in adulthood: a dimensional-spectrum conceptualization and its implications for DSM-V. AB - Mental disorders involving antisocial behavior and substance use are genetically linked and vary continuously. The authors present a review and integrative conceptualization of these observations in terms of a dimensional and hierarchically organized externalizing spectrum. As a foundation for this conceptualization, the authors introduce a quantitative, model-based approach to comparing categorical and continuous conceptions of psychopathology and apply this approach in an empirical study of patterns of comorbidity among externalizing disorders as defined in the Diagnostic and Statistical Manual of Mental Disorders. The authors present evidence that comorbidity among externalizing disorders is best modeled by an underlying normally distributed continuum of risk for multiple disorders within the externalizing spectrum. The authors conclude by discussing implications of the externalizing spectrum conceptualization for classification of disorders in the upcoming 5th edition of the Diagnostic and Statistical Manual of Mental Disorders. PMID- 16351378 TI - Dimensional models for research and diagnosis: a current dilemma. AB - This commentary focuses on the current dilemma regarding the application of dimensional models for research and diagnosis. Four articles are used to review the positive contributions of dimensional approaches. Although categorical approaches are integral in deciding the characteristics of the group to be studied (i.e., development of inclusion/exclusion criteria), the generation of cutpoints for categorical classification is, in reality, working with dimensional data. Ultimately, new methodological strategies need to be incorporated that address both categorical and dimensional aspects of the overall diagnostic framework. These refinements will be vital in determining the extent and reality of co-occurrence of disorder and the determination of boundaries across specific disorders. PMID- 16351379 TI - Clinical utility: a prerequisite for the adoption of a dimensional approach in DSM. AB - A potential obstacle to implementing dimensional representations in the Diagnostic and Statistical Manual of Mental Disorders (DSM) is lack of data about clinical utility and user acceptability. Adopting a dimensional approach would likely complicate medical record keeping, create administrative and clinical barriers between mental disorders and medical conditions, require a massive retreating effort, disrupt research efforts (e.g., meta-analyses), and complicate clinicians' efforts to integrate prior clinical research using DSM categories into clinical practice. Efforts to empirically demonstrate the clinical utility of dimensional alternatives should be a prerequisite for their future implementation in order to establish that their advantages outweigh the disadvantages. Approaches to promote user acceptability and the development of an empirical database include dimensionalizing existing DSM categories and including research dimensions in the DSM appendix. PMID- 16351380 TI - Dimensional models of psychopathology: research agenda and clinical utility. AB - This commentary emphasizes the implications for a mental health research agenda that stem from the papers in this special section on dimensional models of psychopathology. These include the need to extend dimensional models to a wider range of psychopathology; the relationship of the dimensions described in these papers, largely based on symptom and self-report measures, to findings from current research in genetics, neuroimaging, and other domains of neuroscience; the need for new scales that can assess the entire range of relevant dimensions with modern psychometric techniques; and ways to employ these dimensions in applied clinical situations. It is concluded that hierarchical dimensional models offer powerful ways of organizing our thinking about psychopathology and will serve to guide many promising avenues of future research. PMID- 16351381 TI - A genetically informed study of marital instability and its association with offspring psychopathology. AB - Parental divorce is associated with a number of emotional and behavioral problems in young-adult offspring, but theoretical and empirical considerations suggest that the relation may be partially or fully accounted for by passive gene environment correlation or environmental selection characteristics. The current study used the Children of Twins Design to explore whether shared environmental or genetic factors confound the relationship between parental marital instability and measures of psychopathology. Comparisons of the offspring of adult twins in Australia on 3 factors of abnormal behavior, including drug and alcohol, behavioral, and internalizing problems, suggest that environmental influences associated with divorce account for the higher rates of psychopathology. The results are consistent with a causal connection between marital instability and psychopathology in young-adult offspring. PMID- 16351382 TI - Etiological contributions to heavy drinking from late adolescence to young adulthood. AB - The authors examined genetic and environmental contributions to stability and change in heavy drinking from late adolescence to young adulthood in a sample of 1,152 twin pairs. In men, heavy drinking was similarly heritable at ages 17 (h2=.57) and 20 (h2=.39), and its stability owed primarily to common genetic factors. In women, heavy drinking was less heritable than in men at ages 17 (h2=.18) and 20 (h2=.30) and its stability was primarily due to enduring shared environmental influences. P3 amplitude, an event-related brain potential marker of alcoholism risk, was less predictive of heavy drinking in women than in men, providing further support for the proposition that biological factors have less impact on heavy drinking in young adult women than in young adult men. PMID- 16351383 TI - Working memory impairments in schizophrenia: a meta-analysis. AB - Working memory (WM) deficit is a cardinal cognitive symptom of schizophrenia, but the differences among the tasks and measures used to assess WM make it difficult to compare across studies. The authors conducted a meta-analytic review to address 3 major questions: (a) Do patients with schizophrenia show WM deficits across diverse methodology; (b) Is WM deficit supramodal; and (c) Does the WM deficit worsen with longer delays? The results indicate that significant WM deficit was present in schizophrenia patients in all modalities examined. Increasing delay beyond 1 s did not influence the performance difference between schizophrenia patients and healthy control participants in WM. These results suggest that WM deficit in schizophrenia is modality independent and that encoding and/or early part of maintenance may be problematic. PMID- 16351385 TI - Emotion context insensitivity in major depressive disorder. AB - The present study tested 3 competing views of how depression alters emotional reactivity: positive attenuation (reduced positive), negative potentiation (increased negative), and emotion context insensitivity (ECI; reduced positive and negative). Normative and idiographic stimuli that elicited happy, sad, and neutral states were presented to currently depressed, formerly depressed, and healthy control individuals while experiential, behavioral, and autonomic responses were measured. Currently depressed individuals reported less sadness reactivity and less happiness experience across all conditions than did the other participants, and they exhibited a more dysphoric response to idiographic than to normative stimuli. Overall, data provide partial support for the positive attenuation and ECI views. Depression may produce mood-state-dependent changes in emotional reactivity that are most pronounced in emotion experience reports. PMID- 16351384 TI - Conjoint developmental trajectories of young adult alcohol and tobacco use. AB - Developmental and etiological advances have set the stage for considering trajectories of problem behavior across the life course, but little work thus far addresses co-occurring problem behavior trajectories. Although recent work characterizes drinking and smoking trajectories, none has explored the course of concurrent drinking and smoking. Using panel data from the Monitoring the Future Project (N=32,087), the authors applied growth mixture modeling to 4 waves of heavy drinking and smoking in a young-adult sample. The authors extracted a single latent group membership factor from heavy drinking and smoking. Associations between trajectory classes and risk factors were relatively unique to the substance being predicted. The association of smoking with alcohol expectancies and delinquency appeared to exist by virtue of smoking's comorbidity with drinking. PMID- 16351386 TI - Situational correlates of abstinence self-efficacy. AB - Little is known about the situations that are associated with changes in self efficacy during an attempt to quit smoking. In this study, 214 smokers used palmtop computers to record momentary self-efficacy ratings and situational context during a quit attempt. Higher urge to smoke and negative affect were associated with reduced self-efficacy. Although alcohol and coffee consumption are associated with heightened lapse risk, they were unrelated to abstinence self efficacy. Individuals with low baseline self-efficacy generally reported lower self-efficacy across situations, but these differences were more pronounced under conditions of high urge and negative affect. These results suggest that self efficacy may be reactive to affect-motivational states during a quit attempt. Whether these influences represent cognitive biases or objective risk assessments is not known. PMID- 16351387 TI - Dynamic self-efficacy and outcome expectancies: prediction of smoking lapse and relapse. AB - According to social learning models of drug relapse, decreases in abstinence self efficacy (ASE) and increases in positive smoking outcome expectancies (POEs) should foreshadow lapses and relapse. In this study, the authors examined this hypothesis by using ecological momentary assessment data from 305 smokers who achieved initial abstinence from smoking and monitored their smoking and their ASE and POEs by using palmtop computers. Daily ASE and POEs predicted the occurrence of a 1st lapse on the following day. Following a lapse, variations in daily ASE predicted the onset of relapse, even after controlling for concurrent smoking. ASE and POEs generally neither mediated nor moderated each other's effects. These data emphasize the role of dynamic factors in the relapse process. PMID- 16351388 TI - Adolescent girls' interpersonal vulnerability to depressive symptoms: a longitudinal examination of reassurance-seeking and peer relationships. AB - A transactional, interpersonal framework involving adolescents' reassurance seeking and peer experiences may be useful for understanding the emergence of gender differences in depression prevalence during the adolescent transition. Sociometric nominations of peer acceptance/rejection and ratings of friendship quality provided by adolescents and their friends were used to measure peer experiences among 6th-8th-grade adolescents (N=520) over 3 annual time points. After controlling for age and pubertal development, significant but small prospective effects offered mixed support for hypotheses: (a) depressive symptoms and negative peer relations predicted increasing levels of girls' reassurance seeking; (b) initial levels of reassurance-seeking and depressive symptoms predicted deteriorating friendship quality among girls and low friendship stability, respectively; and (c) reassurance-seeking combined with poor peer experiences predicted increases in girls' depressive symptoms. PMID- 16351389 TI - Induced mood change and dysfunctional attitudes in remitted bipolar I affective disorder. AB - This study investigated the possibility that, in remitted bipolar I affective disorder, dysfunctional attitudes are mood-state dependent. Participants were 120 individuals with remitted bipolar I disorder, remitted unipolar depression, or no history of affective disorder. The Dysfunctional Attitudes Scale (DAS; Weissman, 1979) was completed before and after positive or negative mood challenge. Following mood increase, the bipolar group changed significantly less in DAS total score than did the other 2 groups, and in goal-striving and achievement attitudes relative to the unipolar group. These findings did not provide clear support for the mood-state dependency theory in bipolar disorder, arguing instead for the presence in bipolar I disorder of dysfunctional cognitions that show characteristic resilience in the face of minor positive mood increase. PMID- 16351390 TI - Blunted cortisol response to awakening in mild to moderate depression: regulatory influences of sleep patterns and social contacts. AB - Cortisol is elevated during severe depression. However, some studies of outpatients suggest reduced cortisol levels, either basal or poststress. More definite evidence of this phenomenon is needed, and correlates that may explain the disparate findings should be identified. Women from the community (37 depressed and 36 nondepressed) completed electronic diaries in order to help researchers assess the cortisol awakening response (CAR), sleep, and social contacts. Depressed women had a blunted CAR compared with nondepressed women. Among the nondepressed but not among depressed women, time of waking, and number of social contacts (especially positive ones) were independently associated with CAR. These psychosocial factors may contribute to a normal CAR, but their regulatory influence may become disrupted during mild to moderate clinical depression. PMID- 16351391 TI - Executive functions and ADHD in adults: evidence for selective effects on ADHD symptom domains. AB - Dual-process models of attention-deficit/hyperactivity disorder (ADHD) suggest that both executive functioning and regulatory functions (e.g., processing speed) are involved and that executive function weaknesses may be associated specifically with symptoms of inattention-disorganization but not hyperactivity impulsivity. Adults aged 18-37 (105 with ADHD, 90 controls) completed a neuropsychological battery. The ADHD group had weaker performance than did the control group (p<.01) on both executive and speed measures. Symptoms of inattention-disorganization were uniquely related to executive functioning with hyperactivity-impulsivity controlled. Inattention was associated with slower response speed, and hyperactivity-impulsivity with faster output speed. Results were not accounted for by IQ, age, gender, education level, or comorbid disorders. Findings are discussed in terms of developmental and dual-process models of ADHD leading into adulthood. PMID- 16351392 TI - A taxometric investigation of unipolar depression in the national comorbidity survey. AB - Ongoing debate has questioned whether unipolar depression is a dimensional or categorical phenomenon. Although past studies using taxometric methods have supported a dimensional interpretation, each has suffered from methodological limitations. The present study was designed to overcome these limitations through reanalysis of the National Comorbidity Survey. Two indicator sets were constructed from the depression-relevant questions of the Composite International Diagnostic Interview. Participants who endorsed the lifetime occurrence of significant depressed mood or anhedonia (n=4,577) were submitted to 2 nonredundant taxometric procedures (maximum eigenvalue and means above minus below a cut), additional consistency tests, and recently developed simulation techniques. All results converged on a dimensional solution. The implications of these findings on assessment, treatment, and research design are discussed. PMID- 16351393 TI - Distress tolerance as a predictor of early treatment dropout in a residential substance abuse treatment facility. AB - A large percentage of individuals who enter residential substance abuse treatment drop out before completing treatment. Given that early treatment dropout places individuals at an increased risk for relapse, identifying the mechanisms underlying treatment dropout would have several important theoretical and clinical implications. In the current study, the authors examined levels of psychological and physical distress tolerance as a predictor of early treatment dropout in a residential substance abuse treatment facility. In a sample of 122 individuals entering a residential substance abuse treatment facility, level of psychological distress tolerance was predictive of early treatment dropout above and beyond relevant self-report variables. There was no relationship between physical distress tolerance and early treatment dropout. Implications for future studies and treatment development or modification are discussed. PMID- 16351394 TI - Does poor vagal tone exacerbate child maladjustment in the context of parental problem drinking? A longitudinal examination. AB - Children's vagal tone was examined as a vulnerability factor in the longitudinal relation between mothers' (MPD) and fathers' (FPD) problem drinking and children's adjustment. At T1, MPD and FPD were examined, and children's vagal tone was assessed. Two years following initial participation, child adjustment problems were evaluated. A lower level of vagal tone was a vulnerability factor for internalizing problems associated with MPD and FPD and for externalizing difficulties associated with MPD. In the context of a higher level of MPD or FPD, a lower level of vagal tone was a vulnerability factor for increases in externalizing problems over time. Results are supportive of the importance of biopsychosocial perspectives in which child characteristics interact with family risk to predict psychopathology. PMID- 16351398 TI - Charles G. Gross: Award for Distinguished Scientific Contributions. PMID- 16351399 TI - Processing the facial image: a brief history. AB - The study of the neural basis of face perception is a major research interest today. This review traces its roots in monkey neuropsychology and neurophysiology beginning with the Kluver-Bucy syndrome and its fractionation and then continuing with lesion and single neuron recording studies of inferior temporal cortex. The context and consequence of the discovery of inferior temporal neurons selective for faces is described and current lines of research on inferior temporal cortex and face processing in both monkeys and humans are outlined. PMID- 16351400 TI - Douglas L. Medin: Award for Distinguished Scientific Contributions. PMID- 16351401 TI - Robert S. Siegler: Award for Distinguished Scientific Contributions. PMID- 16351402 TI - Children's learning. AB - A new field of children's learning is emerging. This new field differs from the old in recognizing that children's learning includes active as well as passive mechanisms and qualitative as well as quantitative changes. Children's learning involves substantial variability of representations and strategies within individual children as well as across different children. The path of learning involves the introduction of new approaches as well as changes in the frequency of prior ones. The rate and the breadth of learning tend to occur at a human scale, intermediate between the extremes depicted by symbolic and connectionist models. Learning has many sources; one that is particularly promising for educational purposes is self-explanations. Overall, contemporary analyses show that learning and development have a great deal in common. PMID- 16351404 TI - Karen a. Matthews: Award for Distinguished Scientific Applications of Psychology. PMID- 16351405 TI - Psychological perspectives on the development of coronary heart disease. AB - Psychological science has new opportunities to have major input into the understanding of the development of coronary heart disease. This article provides an overview of advances in understanding the etiology of heart disease, recently applied technologies for measuring early stages of heart disease, and an accumulating base of evidence on the psychosocial predictors of clinical and subclinical coronary disease. It suggests that it is now time to develop conceptual models that integrate our understanding of separate psychosocial predictors. It offers and provides preliminary evidence for a model that integrates socioeconomic status, environmental stress, and person-level factors from a life span perspective. PMID- 16351407 TI - Albert Jan (Ap) Dijksterhuis: Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 16351408 TI - Gunther Knoblich: Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 16351409 TI - Robert F. Krueger: Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 16351410 TI - Hendree E. Jones: Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 16351411 TI - Fredrick p. Morgeson: award for distinguished scientific early career contributions to psychology. PMID- 16351412 TI - Russell A. Poldrack: Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 16351415 TI - Margaret Beale Spencer: Award for Distinguished Senior Career Contributions to Psychology in the Public Interest. PMID- 16351416 TI - Crafting identities and accessing opportunities post-Brown. AB - Decades following Brown v. Board of Education (1954), issues regarding the effects of skin color, poverty, and racial differences in the availability of protective factors persist. For a multiethnic sample of mainly African American (56%), female (69%), and high-achieving (65%) youths, a dual-axis model of vulnerability is used to compare four groups of youths identified as high, masked, low, and undetermined vulnerability on the basis of the presence of risk and protective factors. Risks included level of poverty, number of parents in home, and skin-color pigmentation. Protective factors included youths' perceptions of school climate, parental monitoring, and consonance between skin pigmentation and preferred skin color. The author suggests that 50 years post Brown, issues concerning White privilege, color stereotyping, power discrepancies, and economic disparities maintain "invisible" and persistent hurdles for vulnerable youths who vary on available protective factors. Findings confirm the impact of vulnerability on psychosocial and achievement outcomes. PMID- 16351417 TI - Daniel Dodgen: Award for Distinguished Early Contributions to Psychology in the Public Interest. PMID- 16351419 TI - Gail S. Goodman: Award for Distinguished Contributions to Research in Public Policy. PMID- 16351422 TI - John C. Norcross: Award for Distinguished Contributions to Education and Training. PMID- 16351423 TI - The psychotherapist's own psychotherapy: educating and developing psychologists. AB - In this article, the author synthesizes 25 years of his research on the personal therapy of mental health professionals. The author summarizes the high prevalence of personal treatment, its use by seasoned practitioners, its personal and professional goals, its typically positive outcomes, and its lasting lessons concerning the practice of psychotherapy. Particular attention is devoted to the therapist's selection of a personal therapist and the conduct of treatment with patients who are themselves psychotherapists in comparison with patients who are laypersons. The cumulative results indicate that personal therapy is an emotionally vital, interpersonally dense, and professionally formative experience that should be central to the development of health care psychologists. PMID- 16351424 TI - Irma Serrano-Garcia: Award for Distinguished Contributions to Education and Training. PMID- 16351426 TI - Cal D. Stoltenberg: Award for Distinguished Contributions of Applications of Psychology to Education and Training. PMID- 16351427 TI - Enhancing professional competence through developmental approaches to supervision. AB - There has been a recent increase in interest in defining and describing the competencies for professional practice in psychology. Perhaps the most important mechanism for enabling the acquisition of competencies is the process of supervision. This article takes the position, based on a review of relevant research in supervision and the author's experiences, that developmental theories of supervision have stimulated considerable research and provide a direction for future work in research and practice. It is argued that the most detailed of these theories, the Integrated Developmental Model, provides a useful framework for understanding how supervisees change over time and how various supervision environments (broadly) and supervision interventions (specifically) can enhance or detract from the development of professional competencies. PMID- 16351429 TI - George M. Slavich: Psi Chi/APA Edwin B. Newman Graduate Research Award. PMID- 16351432 TI - Gail S. Goodman: Award for Distinguished Professional Contributions to Applied Research. PMID- 16351433 TI - Wailing babies in her wake. AB - The scientific study of child witnesses has influenced both developmental science and jurisprudence concerning children. Focusing on the author's own studies, 4 categories of research are briefly reviewed: (a) children's eyewitness memory and suggestibility; (b) memory for traumatic events in childhood; (c) disclosure of child sexual abuse; and (d) experiences of child victim/witnesses within the legal system. Implications for psychology and for legal practice are discussed. PMID- 16351435 TI - Edward A. Wise: Award for Distinguished Professional Contributions to Independent or Institutional Practice in the Private Sector. PMID- 16351436 TI - Effectiveness of intensive outpatient programming in private practice: integrating practice, outcomes, and business. AB - The development of a model for treating acutely depressed patients on a frequent basis in an independent practice setting is described. Strategies to collaborate with managed care organizations, employee assistance programs, and local provider networks to recruit these patients are outlined. The patients treated in the intensive outpatient program described in this article were primarily depressed, were more acutely distressed than national and local inpatient samples, were often in suicidal crises, and had multiple comorbidities. Nonetheless, they demonstrated significant pre- and posttest improvements on a variety of client- and clinician-rated measures. A dose-response curve further illustrated that significant improvement in depressive symptoms was predictable. Private practitioners can expand their services through the development, implementation, delivery, management, and evaluation of mental health services that respond to market dynamics. PMID- 16351438 TI - Morgan T. Sammons: Award for Distinguished Professional Contributions to Practice in the Public Sector. PMID- 16351439 TI - Psychology in the public sector: addressing the psychological effects of combat in the U.S. Navy. AB - The response of military psychology in times of war or other great public crises may presage the success of the profession in less perilous times. The ability of public-sector psychologists to provide assistance and improve the common welfare during conflict or turmoil is generally followed by an increased demand for psychological services. This likely reflects the success of the psychological response during those crises, and it underscores the fact that psychological consequences of war or disaster require both immediate clinical attention and long-term policy development. The U.S. Navy serves as a model for public-sector psychological service provision. A brief history of Navy psychology is provided, followed by an examination of how Navy psychologists are responding to the issues raised by the current conflict in the Middle East and the problems associated with stigma in the treatment of posttraumatic stress disorder. PMID- 16351441 TI - Renee E. DeRouin: APA/APAGS Award for Distinguished Graduate Student in Professional Psychology. PMID- 16351444 TI - Gary B. Melton: Award for Distinguished Contributions to the International Advancement of Psychology. PMID- 16351445 TI - Building humane communities respectful of children: the significance of the convention on the rights of the child. AB - The greatest promise of international human rights law is in stimulating thoughtful self-examination by those who strive to do what is right. Perhaps more than any other instrument, the Convention on the Rights of the Child (CRC; 1989) offers the opportunity to use a global moral consensus to transform life at the neighborhood level in order to promote "the full and harmonious development of...[the child's] personality" (CRC, 1989, preamble). The CRC also presents a normative framework to guide psychologists' advocacy on behalf of children. PMID- 16351446 TI - Charles D. Spielberger: Award for Distinguished Contributions to the International Advancement of Psychology. PMID- 16351448 TI - Eduardo Almeida: International Humanitarian Award. PMID- 16351449 TI - [Pharmacists commitment to pharmacotherapeutic quality and patient safety]. PMID- 16351450 TI - [Medical gases, new medications]. PMID- 16351451 TI - [Improving adequacy for drug use and effects in geriatric centers using an intervention program]. AB - OBJECTIVE: To evaluate the impact of a program for increasing adequacy of drug prescriptions in a group of nursing homes 2 years after implementation. METHOD: This quasi-experimental before and after study was carried out with a control group in various centers. It included all outpatient prescriptions, individualized by centre, during the study period (reference: year 2001 and post intervention: year 2003) with 107/118 nursing homes from the Region Sanitaria del Barcelones Norte y Maresme (Barcelona) included in each period. After an initial situation analysis, centers were assigned to two different groups: intervention group (n = 32) and control group (n = 75/86). Number of residents: 4,798/5,816 (years: 2001/2003). Actions accomplished in the intervention group were: a) presentation letter, b) informative interview c) management control and d) monitorization with follow-up interviews. Quantitative (total cost of drugs among residents) and qualitative indicators (high intrinsic pharmacologic value drug use, generic drug use and super-night diaper use) were established as measurement units, together with relative use values. RESULTS: In the intervention group, cost contention was evident. Cost per resident was 1,671.89 EUR +/- 458.33 EUR in the reference period and 1,821.22 EUR +/- 311.88 EUR during the post-intervention period (inter-annual increment of 8.9% vs. 19.5% compared to the control group; p = 0.002). Number of packages per resident showed an inter-annual increment of 2.5% and 11.4% (p = 0.001) respectively. An increased use of generic drugs (7.9% and 18.4%), antiasthmatic agents, omeprazol and recommended nonsteroidal antiinflammatory agents was observed; while there were no differences in the use of antidepressants and antibiotics. CONCLUSIONS: The preliminary results have shown improved efficiency of pharmaceutical prescriptions in nursing homes included in the intervention group. The methodology used appears to be appropriate for promoting rational use of drugs and improving prescription quality. PMID- 16351452 TI - [Pharmaceutical care in a social-healthcare center]. AB - OBJECTIVE: To reflect the activity of the pharmacy department within the nursing home s multidisciplinary team, and to assess care improvements received by residents, including a better use of drugs. METHOD: In addition to pharmacotherapeutical guidelines and a standardized work procedure listing generic and therapeutic replacements to be implemented, selected procedures were specified in order to ensure a rational use of drugs and to increase the quality of care received by residents in our center. Such procedures will be performed by the center s multidisciplinary team and will focus on caring the caring for so called geriatric syndromes. RESULTS: After an assessment of treatments for 125 patients and a 3-month follow-up, 10 interventions per patient were performed, out of which 4.74 were pharmacist interventions and 5.26 were to increase quality of care. CONCLUSIONS: The presence of the pharmacy department within the multidisciplinary team ensures a better use of drugs and a safe delivery system. Its activity contributes to improve quality of care, most notably procedures regarding nutritional status, and the prevention and treatment of pressure ulcers. PMID- 16351453 TI - [A project for the implementation of a unit-dose drug dispensation system in an intensive care unit]. AB - OBJECTIVE: To describe the implementation of a unitary dose drug dispensation system (UDDDS) with computerized medical orders in an intensive care unit (ICU) including 10 multi-purpose offices, and to obtain a medication error index as an indicator of the process quality. METHOD: A UDDDS with computerized medical orders for intensive care was defined. By consensus among nurses, intensivists and pharmacists, the administration of high-risk drugs by perfusion or through a gastric tube was protocolized, and computerized medical orders were adapted to ICU dynamics, with both fluid therapy and enteral and parenteral nutrition becoming fully integrated. A prospective observational 8-month study with 15 cross-sectional time points was performed to estimate the overall error index and mean error per drug use process stage. The error index is estimated by dividing the number of errors into error opportunities, and is expressed as a percentage. RESULTS: Computerized medical orders favored compliance with consensus protocols defined in software programs at the pharmacy department, even though the degree of adhesion degree was not quantitized. They also allowed a validation of all medical prescriptions by a pharmacist before dispensation. The total number of errors detected during the study period was 86. Error opportunities were 26,695, and the overall error index was 0.32%. During the study an error occurred every 312.5 error opportunities. PMID- 16351454 TI - [A study of initial antiretroviral therapy effectiveness, survival and safety in HIV+ patients]. AB - OBJECTIVE: To study initial antiretroviral therapies indicated for HIV-infected patients during the 2001-2003 period regarding effectiveness, survival and safety. METHOD: A descriptive, retrospective study of clinical and drug-related variables of naive HIV-infected patients through pharmacotherapeutic history. RESULTS: Mean CD4+ lymphocytes counts were 209.6 cells/mm3. Pneumonia by Pneumocystis carinii was the most commonly found condition at antiretroviral treatment onset. Most commonly used therapies included those based on a non nucleoside reverse transcriptase inhibitor (NNRTI) combined with two nucleoside reverse transcriptase inhibitors (NRTIs). The longest mean survival was achieved by using combinations of three nucleoside reverse transcriptase inhibitors. The primary reason for initial antiretroviral therapy discontinuation were adverse effects, with stavudine exhibiting the poorest tolerability. CONCLUSIONS: Therapies based on non-nucleoside reverse transcriptase inhibitors and protease inhibitors (PIs) have shown similar effectiveness to increase CD4+ cell counts. Regarding viral load decreases, protease inhibitors were most effective. Therapies using three nucleoside reverse transcriptase inhibitors resulted in peak survival. PMID- 16351455 TI - [Stability and in vitro activity of voriconazole eyedrops at a concentration of 3 microg/mL]. AB - OBJECTIVE: To assess the stability and activity of voriconazole 3 microg/mL eyedrops as prepared for use against amphotericin B- and fluconazole-resistant fungal endophthalmitis. METHOD: Stability (concentration using UV spectrophotometry; pH, osmolarity, and particle formation) and sterility were analyzed under various preservation conditions--room temperature (22-24 degrees C) or refrigerated (2-8 degrees C). The preparation's in vitro efficacy was analyzed using the standard National Committee for Clinical Laboratory Standards method for 30 days. RESULTS: Voriconazole concentrations were found to be within limits allowed by the United Stated Pharmacopeia (90-115%). pH (room temperature: 6.96-7.60; refrigerated: 6.84-7.21) and osmolarity (room temperature: 265-284 mOsm/l; refrigerated: 270-285 mOsm/l) remained within eye physiological ranges throughout the study under the analyzed conditions. The preparation s antifungal activity remained stable during the first three weeks. CONCLUSIONS: The voriconazole 3 microg/mL eyewash preparation remained stable, sterile and with full antifungal activity for 21 days when stored both at room temperature and under refrigeration conditions. PMID- 16351457 TI - [Role of a clinical pharmacist in a consensus conference on therapeutic recommendations for fungal infection]. PMID- 16351456 TI - [Current research status in pharmaceutical care]. AB - OBJECTIVE: A systematic review of studies on pharmaceutical care research from June 1999 to June 2004 was carried out. METHOD: Medline, Current Contents, Cochrane Library, IDIS, and Teseo were used as data sources. Works were categorized according to evidence levels and recommendation grades in clinical practice guidelines. The JADAD method was used for quality quantification. RESULTS: In all, 129 references were found; 19.4% (n = 25) were randomized studies; 2.3% (n = 3) had blinded assessors; losses to follow-up were documented in 7.8% (n = 10); 4.7% (n = 6) had a Jadad score= 3; 8.5% (n = 11), 14.7% (n = 19), 5.4% (n = 7), and 20.9% (n = 27) had evidence levels Ia, Ib, IIa, and IIb, respectively; 44.2% (n = 57) and 6.2% (n = 8) had levels III and IV, respectively; 23.3% (n = 30) had a recommendation grade A; 26.4% (n = 34) had a grade B; 44% (n = 55) had C and 6.2% (n = 8) had D. Most common study types included: descriptive (39.5%), trials including patients (32.6%), and reviews (17.8%); 59.5% of reviewed clinical trials were controlled, randomized clinical trials (CRCTs). Studies were double-blind in 7.1% of cases. Discontinuations and exclusions were recorded in 23.8% of cases. Randomization was appropriate in 11.9% of cases; 14.3% of clinical trials had 3 points, and 85.7% of studies were of poor quality. CONCLUSIONS: Work methodology should be more rigorous. The use of universally accepted methods is needed to enhance the quality of studies (Jadad system, Consort list). The performance of observational, prospective, multicenter investigations allowing the effectiveness and efficiency of pharmaceutical care to be measured would be most beneficial. Works should measure health-related quality of life (SF-36 questionnaire) and patient satisfaction. PMID- 16351458 TI - [Miliary tuberculosis following treatment with adalimumab]. PMID- 16351459 TI - [Quality of clinical trials provided in GINF]. PMID- 16351460 TI - Genomic instability and colorectal cancer. PMID- 16351461 TI - Tumoral response factors after radiofrequency ablation of hepatocellular carcinoma in cirrhotic liver. AB - OBJECTIVE: Hepatocellular carcinoma (HCC) ablation by radiofrequency (RFA) is a novel technique with a great variety of methods whose efficacy and predictive factors have not been completely studied. Some of the main predictive factors in this type of treatment are analyzed in the present study. PATIENTS AND METHODS: Ninety-three patients with hepatocellular carcinoma over cirrhosis, and with no indication for surgical resection were treated by RFA. Two different types of electrodes were used for RFA (refrigerated-"Cool-Tip" and perfusion with saline solution, the approach was percutaneous, by laparoscopy or laparotomy. RESULTS: Overall survival at 1, 2 and 3 years was 88, 81, and 76%, with a free-disease survival (FDS) of 66, 31 and 17%, respectively. For tumors less than 3 cm, FDS at 1,2 and 3 years was 74, 44 and 30%, while for more than 3 cm in size FDS was 55, 12 and 0% (p = 0.02). FDS for HCC with one nodule was 70, 36 and 22%, and for more than one nodule it decreased to 50, 17 and 0% at 1, 2 and 3 years, respectively (p = 0.07). Surprisingly, the method employed for RFA has a main influence in FDS, with 0% at 3 years for perfusion electrodes and 26% for cool tip electrodes at the same period. CONCLUSIONS: In this series, overall survival at three years was relatively high; however, tumoral size, number of nodules and RFS method were independent variables associated with disease-free survival. PMID- 16351462 TI - Association between angiogenesis soluble factors and disease progression markers in chronic hepatitis C patients. AB - OBJECTIVES: Our objectives were to compare angiogenesis soluble factor (ASF) levels in chronic hepatitis C (CHC) patients and healthy individuals, and to investigate potential associations between ASF levels and both histological and biochemical markers of disease progression. METHOD: Thirty-six patients (69% males) positive for HCV-RNA by PCR analysis were included in the study. All patients underwent liver biopsy before treatment. Serum levels of vascular endothelial growth factor (VEGF), soluble Flt-1 and Flk-1 receptors, placental growth factor (PlGF), angiopoietin-2 (Ang-2) and soluble Tie-2 receptor were determined by ELISA. Fifteen healthy subjects were used as controls. RESULTS: In comparison to healthy individuals, CHC patients showed significantly increased serum levels of proangiogenic factors PlGF (22 +/- 5 vs. 18 +/- 8 pg/ml; p < 0.05), Ang-2 (1265 +/- 385 vs. 833 +/- 346 pg/ml; p < 0.005) and sFlt-1 (95 +/- 22 vs. 72 +/- 14 pg/ml; p < 0.0001). Interestingly, in CHC patients serum levels of VEGF and Tie-2 correlated with grade of inflammation, PlGF correlated with stage of fibrosis, and Flt-1 and Flk-1 correlated with serum transaminase levels (p < 0.05 in all cases). CONCLUSIONS: CHC patients showed increased serum levels of ASF, and a significant correlation was shown between serum levels of selected ASFs and grade of inflammation, stage of fibrosis, and transaminase levels. PMID- 16351463 TI - Acute appendicitis in an incarcerated crural hernia: analysis of our experience. AB - INTRODUCTION: The finding of the vermiform appendix within a crural hernia (Amyand s hernia) is a rare entity whose incidence is not described in the literature. OBJECTIVE: The aim of this study was to report our hospital s experience in this kind of pathology. MATERIAL AND METHODS: Between 1993 and 2004, 4,572 acute appendicitis and 372 incarcerated crural hernia cases have been operated on in our hospital. We studied 6 cases of incarcerated crural hernia with vermiform appendix inside. We analyzed in retrospect the following parameters: age, sex, personal history, clinical manifestations, preoperative diagnosis, surgical technique, mean hospital stay, and outcome. RESULTS: All patients were women with a mean age of 78.8 years. Most frequent clinical manifestations included pain and a mass in the right inguinocrural region, of variable intensity and duration. No clinical, laboratory, or radiographic signs help in reaching a correct preoperative diagnosis. General anesthesia and a crural approach are used in most surgical operations. In all cases an appendectomy was performed via the hernia sac, thus proving the presence of acute appendicitis in four of them (66.67%). A prosthetic mesh was used in 3 cases, and one case of wound infection was found. In the other cases we sutured the hernia ring using prolene. CONCLUSION: The finding of the appendix in an incarcerated crural hernia is a rare entity in old women that is difficult to diagnose preoperatively. Treatment includes appendectomy and herniorraphy. The use of prosthetic mesh is controversial. PMID- 16351464 TI - Peritoneal carcinomatosis of colorectal origin. Current treatment. Review and update. AB - Colorectal cancer is the most frequent tumor of the digestive tract. The high incidence of abdominal dissemination; the poor prognosis of these patients, with median survival consistently ranging from 5 to 9 months in all studies of peritoneal carcinomatosis from colorectal cancer; the failure of adjuvant systemic chemotherapy treatment with a maximal survival of 18 months despite the development of new cytostatic drugs, and new combinations of use, make it crucial to search for and develop new treatment strategies. We review the principles of Sugarbaker s treatment protocol, which involves the combination of maximum cytoreductive radical oncological surgery for the treatment of all macroscopically disseminated disease with maximum perioperative intraperitoneal intensification chemotherapy to treat residual microscopic disease. We present the results of several scientific papers, all of them phase II studies with more than 10 patients treated, published in the medical literature by the main groups working in this line of treatment, together with the only phase III study reported and published so far, and finally the results of a recently reported retrospective international multicenter study. With this new alternative therapeutic approach, overall mean survival is 40% at 36 months, and 20% at 5 years. Based on these results, this new therapeutic approach is proposed as the treatment of choice for these unfortunate patients. PMID- 16351465 TI - APC and chromosome instability in colorectal cancer. AB - Colon cancer is a common disease that can be sporadic or familial. An inactivated adenomatous polyposis coli (APC) suppressor gene is found in over 80% of colorectal tumors, this being an early alteration in the development of adenomatous polyps. APC function is not only critical for tumor initiation and progression, and chromosome instability (CIN) is another characteristic dependent at least partly on APC mutations. PMID- 16351466 TI - Laparoscopic colectomy for primary colonic lymphoma. AB - Primary colorectal lymphoma is an infrequent disease of unknown origin and with a growing incidence. Differentiation be-tween primary lymphoma and secondary colorectal involvement is of great therapeutic and prognostic importance. The diagnosis must always be based on histological findings. Different forms of presentation have been described, though all are nonspecific. While full agreement on the best management approach for colorectal lymphoma is lacking, surgical resection is usually the treatment of choice. The role of chemotherapy has not been defined. This study reports on a case of primary lymphoma in the cecum, with clinical manifestations in the form of meteorism and early satiety. The diagnosis was established by colonoscopy and biopsy. The absence of lymph node involvement, tumor size, existing cardiopulmonary risk factors, and the fact that a full resection of the malignancy proved possible, with tumor-free resection margins, led us to exclude adjuvant chemotherapy. After one year of follow-up the patient remains disease-free. PMID- 16351467 TI - Choledocho-renal fistula in locally advanced cholangiocarcinoma. PMID- 16351468 TI - [Esophageal lichen planus: clinical, endoscopic, endosonographic and histological findings]. PMID- 16351469 TI - [Giant pancreatic cystadenoma. A very rare entity]. PMID- 16351470 TI - [Submucous rectal mass and acute meningitis: presentation of an anterior sacral meningocele]. PMID- 16351471 TI - [Idiopathic fibrosing pancreatitis as a rare cause of obstructive jaundice]. PMID- 16351472 TI - [Eosinophilia and actinic enteritis due to radiotherapy for prostatic adenocarcinoma]. PMID- 16351473 TI - [Appendiceal mucinous cystoadenocarcinoma: endoscopic diagnosis]. PMID- 16351474 TI - [The clinical importance of the polymorphisms of CD45 gene]. PMID- 16351475 TI - [The use of a simple algorithm to improve proteinuria diagnosis in clinical laboratories]. AB - BACKGROUND AND OBJECTIVE: When proteinuria appears, a differential diagnosis must determine its origin. The object of this work has been to evaluate the results after the laboratory implantation of an algorithm for the screening and diagnosis of proteinuria. MATERIAL AND METHODS: From a total of 30,718 processed urines, a 30 mg/dl or higher protein concentration was obtained in 639, recommending a new sample to confirm and differentiate proteinuria. We received 207, to which total protein, creatinine, albumin and alpha-1-microglobulin were quantified, together with pseudoperoxidase and leukocyte esterase from dipstick. The results were introduced in an expert system (UPES and its application Protis), allowing differentiate hematuria, leukocyturia and proteinuria and suggesting the assessment of other parameters, like IgG, alpha-2-macroglobulin, light chain kappa/lambda, when necessary. RESULTS: From 207 urinalysis assayed for selective proteinuria, 39 were normal, 96 were classified as primary glomerulopathy, 26 as secondary glomerulopathy and 5 as tubulo-interstitial nephropathy. A differential diagnosis of hematuria was made in 58 of these urines. Besides, kappa light chains were detected in a sample from a patient with a normal serum protein graph, which were confirmed by immune fixation. CONCLUSION: With the proposed algorithm, the information obtained from a urine sample increases substantially, allowing detection and differentiation of proteinuria and providing suggestions for the clinical evaluation of the patient. PMID- 16351476 TI - [Venous thromboembolic disease in a rural area of Southern Extremadura (Spain)]. AB - OBJECTIVE: To study the characteristics of the venous thromboembolic disease (VTD) in a rural area of Southern Extremadura (Spain). MATERIAL AND METHODS: Prospective observational study of all patients diagnosed by objective methods of VTD in the only hospital in our area from November 2002 to November 2004. RESULTS: Seventy-six patients were diagnosed of VTD: 35 (46.1%) with pulmonary thromboembolism and 41 (53.9%) with isolated deep vein thrombosis (DVT). The median age was 72.4 years (range 35-94 years), 51 (67.1%) were older than 70 years, and 43 (56.6%) were women. The annual incidence of VTD was 0.63 per 1,000 persons (0.74 per 1,000 persons in women and 0.54 per 1,000 persons in men). Thirty-seven patients (48.7%) had "idiopathic" VTD. After a median follow-up of 5.5 months with anticoagulant therapy, 13 patients (17.1%) had dead. Comorbidity and non "idiopathic" VTD were significantly more frequent in the dead patients. Three patients (3.9%) presented probable recurrence, and all of them died. There were 4 cases (5.3%) of non-fatal severe hemorrhage. Nine patients with isolated DVT did not require hospitalization, and there was no event in the follow-up. CONCLUSIONS: The incidence of VTD may be lower in our area than in other geographical areas. VTD affects more frequently to elderly and women, and almost half of cases are "idiopathic". The mortality is high, and it is related to the existence of comorbidity and non "idiopathic" VTD. The recurrences and the severe hemorrhages are not exceptional during the anticoagulant therapy. The ambulatory treatment of isolated DVT may be an option in selected patients. PMID- 16351477 TI - [Estimation of vitamin supplements and minerals consumption dispensed through pharmacy offices in the province of Las Palmas]. AB - BACKGROUND: Vitamin supplements and minerals consumption (SPM) is increasing in occidental societies due to the growing concern about health by the population. OBJECTIVES: To have a initial approaching to the to SPM consumption in the Province of Las Palmas through 2000 and 2001. To identify SPM proportions that are dispensed as pharmaceutical specialities and those who are sold as parapharmacy products. Finally, to describe the evolution of this consumption throughout a year. METHOD: The information was obtained through the list of the whole pharmaceutical specialities and parapharmacy products through 2000 and 2001 who have at least a vitamin in its composition and/or a mineral. RESULTS: Usually, 297 pharmaceutical specialities and 216 parapharmacy products are currently being sold. Pharmaceutical specialities comprised 65.6% of the whole products sold and within them, vitamins were the most dispensed (41.5%). Regarding parapharmacy products, vitamins and minerals compounds were the products more sold (34.6%). Comparing to 2000, during 2001 there was a statistically significant increase in the consumption of parapharmacy products, remaining without changes the use of pharmaceutical specialities. CONCLUSION: On the basis of sold XX SPM consumption seems to be due mainly by pharmaceutical specialities rather than parapharmacy products. Nevertheless through 2001 there was an increase only in the parapharmacy products. PMID- 16351478 TI - [Herpetic encephalitis in adults: 23 cases]. AB - OBJECTIVE: To study the clinical presentation, diagnosis, treatment and prognosis of the herpetic encephalitis in our environment. MATERIAL AND METHOD: During the past 12 years, 23 adults (age > 15 years) were treated for herpetic encephalitis in our centre. RESULTS: There were 14 males and 9 females. Ages ranged from 18 to 84 years (mean, 52.30 +/- 18.64 years). The hospital stay ranged from 5 to 64 days (mean, 26.74 +/- 15.41 days). Eleven patients were managed in the intensive care unit and nine patients required mechanical ventilation. The most frequent clinical features they were the level of conscience decrease and fever. The lymphocytic pleiocytosis was the most frequent discovery in cerebrospinal fluid. The temporal lobe lesions in the computed tomography scan appeared in 14 patients (61%). Four patients died, seven patients were asymptomatic or with minimum sequels and twelve patients developed they were disabled. CONCLUSIONS: The herpetic encephalitis is an uncommon illness. Intravenous acyclovir is recommended treatment and the corticosteroids use is controverted. The delay in the treatment beginning worsens the prognosis. Less than a third of the patients achieve the functional independence to discharge hospital. PMID- 16351479 TI - [Pulmonary embolism and myxoma of right atrial]. AB - We describe the case of a patient diagnosed of pulmonary embolism, in whom transthoracic echocardiography demonstrated a large mass. It was completely removed and histologic examination showed a very calcification myxoma. This case is of interest because localization and extension of myxoma, the atypical histologic appearance, and in fact the diagnosis and treatment prevented new pulmonary embolism that could be lethal. PMID- 16351480 TI - [Hyperthrombophilia status in a patient without thrombotic events]. AB - Some case reports have been published in the literature about thrombosis associated with several risk factors, especially in hypercoagulability state. Nevertheless, we have not found any description of a case like this of hyperthrombophilia status without thrombotic events. We present a 75 year old woman who was assessed for thrombophilia state on occasion of a deep venous thrombosis which affected to her daughter. Many thrombosis risk factors were detected (13 altogether), both inherited and acquired, arterial and venous, that surprisingly, have not led to any thrombosis event. Its description led us to highlight the risk factors of this patient and to open questions about the present knowledge on etiology and etiopathogenesis of thrombotic phenomena. PMID- 16351481 TI - [Treatment response to lamivudine in chronic hepatitis B patients]. AB - OBJECTIVE: The main objective was to find a biochemistry and virology response in positive and negative HBeAg patients who had been treating with lamivudine. Furthermore, the secondary objective was to make a description about kidney function changes that could be found in patients under treatment with lamivudine. METHOD: Thirty patients with chronic B hepatitis under treatment with lamivudine had been retrospectively studied since November 1999 to March 2004, in a Digestive Unit, in Ciudad Real Hospital. RESULTS: Twenty nine out of 30 patients were included, 8 out of 29 were positive HBeAg, and 21 out of 29 were negative HBeAg. Seven patients had been treated with IFN. Thus, 8 patients had been in need of adefovir treatment after lamivudine trial. The average of treatment had been 23.7 months (3 months-46 months). In fact, none of them were changed from positive HBsAg to negative HBsAg. Nevertheless, we have observed those results after lamivudine treatment. Firstly, 50% positive HBe Ag patients have carried on negative DNA results. Although 25% positive HBe Ag patients had converted to negative HBe Ag. But they did not show any anti HBe. Secondly, 76% negative HBe Ag patients had been suffering a DNA decreased, even though some of them had had a negative DNA results. Thirdly, we found some lamivudine mutation (50% in positive HBe Ag patients and 19% in negative HBe Ag patients). Finally, all the patients with negative DNA had maintained a normal AST and ALT levels. Thus, function renal had been normal under lamivudine trial. CONCLUSION: According to several authors, the response to lamivudine treatment had been adequate in our population. Nevertheless, our study have shown that antiHBe levels had not been suffering any change during lamivudine treatment. In addition, we have found a smaller figures (19%) of lamivudine mutations in negative HBe Ag patients than other studies (25% in patients who were followed up for 1 year). PMID- 16351482 TI - [Bacteremic infection due to Nocardia otitidiscaviarum: case report and review]. AB - We present a case of bacteremic pneumonia caused by Nocardia otitidiscaviarum in a corticodependent COPD. Blood and sputum cultures on Mycobacterial media were positives and identification was done using 16S rDNA sequencing. In this article we review the most relevant communications about Nocardia spp infection and study the strain susceptibility using E-test. PMID- 16351483 TI - [Maple syrup disease: a rare entity that we must know. Review of its dietetic management]. AB - Maple syrup disease is secondary to a deficiency of deshydrogenase complex of a cetoacid of branched-chain. This disease has a recessive autosomic inheritance, with an incidence of 1/200,000 newborns, without differences between male and female. Due to clinical presentation and biochemical response to tiamin, these patients can be classified in five clinical entities: classic, intermediate, intermittent, positive response to tiamin and deficiency of dihydrolipoyl deshydrogenase (E3). In these patients, an increase of seric branched-chain aminoacids is detected, it could be detected by (chromatography) during neonatal period. Valine, isoleucine, and aloisoleucine are increased in serum, orine, and cephaloraquideum liquid by ionic changed chromatography, chromatography of high resolution or high voltage electrophoresis. Patients have two phases in this disease (acute phase and maintaining phase). Objectives in acute phase are based in three topics: to eliminate toxic metabolites, nutritional support and to get anabolism. Utilization of hemodialysis/peritoneal dialysis/blood exchange is one of the first treatments. Dietetic support is the second treatment, with a minimum energy intake and controlling blood levels of aminoacids. Modified dietetic formulas is a main device to treat these patients. PMID- 16351484 TI - [Pain and functional impotence in the lower right extremity]. PMID- 16351485 TI - [Delay in the surgical treatment of lung cancer]. PMID- 16351486 TI - [Delay between diagnosis and surgical treatment of lung cancer in Virgen Macarena Hospital area]. AB - OBJECTIVE: Lung cancer is the most common cause of death due to cancer in the world. Prognosis depends very much on a precocious diagnosis and treatment, being surgery the one that brings the best life expectancy. We studied the time between the diagnosis and the surgical treatment of lung cancer in the hospital area of Virgen Macarena, Sevilla. Making this way a determination of the surgical delay in the treatment. MATERIAL AND METHODS: We analyzed retrospectively the patients who went under surgical treatment in our area between January 2003 and September 2004. And estimating the time in days, between the dates we obtained the first diagnostic specimen of lung cancer in this patients and the date the surgical treatment was performed. RESULTS: We obtained a median of 31 days with a (recorrido intercuartilico) of 60, and a media of 46.16 with a standard deviation of 34.51. CONCLUSIONS: When we compared the results obtained with that we found published, they were similar to those found in the literature. PMID- 16351487 TI - [Clinical and biochemical characteristics of patients with pituitary disease]. AB - OBJECTIVE: Descriptive studies in our country of hypophysis diseases are uncommon. Pituitary adenoma are unfrequent, however this pathology represent 10 12% of brain tumors. Pituitary adenomas are derived of adenohypophysis cells, depending of cellular line, clinical and biochemical characteristics are different. The aim of our study was to describe these characteristics in our patients. MATERIAL AND METHODS: We study 58 patients with diagnosis of pituitary tumor, with regular follow-up in our Hospital, between 1999-2001. The next data were recorded: epidemiological, clinical and biochemical data. RESULTS: Average age at diagnosis was 36.4 years (DS 16.34 years), with a 70.61% of females. The most frequent tumor was prolactinoma and non-functioning adenomas, amenorrhea was the most frequent symptom with the specialist physician (15%) and the general practice physician was the first step in diagnosis protocol. Cerebral image technique was realized in 54 patients, with a 15.5% of normal exploration and 78.6% with lesions, with 24.14% of microadenoma and macroadenoma. Invasive lesions (craniopharyngiomas, germinomas and invasive adenomas) representing a 6.9%. Depression, hypertension and diabetes mellitus were the most frequent diseases in these patients. CONCLUSIONS: Prolactinoma is the most frequent hypophysis pathology. Amenorrhea and/or galactorrhea are the most common symptom. Response to medical and surgical treatment was excellent in prolactinomas. PMID- 16351488 TI - [Home versus inpatient therapy for deep venous thrombosis. A cost-comparative analysis]. AB - AIMS: To compare the home-care management of deep vein thromboses (DVT) by a Home Care Unit (HCU) respect to conventional inpatient treatment. METHODS: Twenty-one patients with a doppler-ecography diagnosis of DVT were managed by the HCU during 2002. In 7 out 13 a concomitant diagnosis of pulmonary embolism (PE) was made by lung scan. Median age was 81 years, 52% were women and all, except one case, showed severe medical concomitant conditions. All patients received low-weight molecular heparin, followed by oral anticoagulants in 3 patients. No patients died and only one was hospitalized briefly due to a poor thrombosis-related pain control. Costs of this patient were added to those of HCU. A comparison was made between ambulatory and hospitalary costs for EP and DVT. Pharmacological treatment costs were calculated for a 10-days period. RESULTS: The length of inhospital stay was 1 day for HCU vs. 8 days (DVT) and 13 days (EP). There was a estimated cost-saving of 1680 per patient. CONCLUSIONS: The management of DVT in patients with serious conditions, can be accomplished safely and in a cost-saving manner by a Home Care Unit. PMID- 16351489 TI - [Outcome of patients with suspected pulmonary thromboembolism and low probability ventilation/perfusion lung scan who receive no long-term anticoagulation]. AB - OBJECTIVE: To know in our area the security of no long-term anticoagulation in patients with suspected pulmonary thromboembolism (PTE) and a low probability ventilation/perfusion (V/Q) lung scan. MATERIAL AND METHODS: Retrospective review of a series of consecutive outpatients with suspected PTE and a low probability V/Q lung scan, according to the modified PIOPED criteria, who receive no long term anticoagulation. RESULTS: Among 38 patients with a low probability V/Q lung scan, 31 (81.6%) did not receive long-term anticoagulation. The median age was 69.1 years (range 26-88 years), and 19 (61.3%) were female. The clinical probability of PTE was moderate in 27 patients (87.1%). Twenty-two patients (71%) had a venous lower extremities echography-doppler negative for deep vein thrombosis (DVT). The median follow-up was 6.3 months (range 3-12 months). There was one case (3.2%; 95% confidence interval, 0.1-16.7%) with demonstrated PTE and DVT, and there was no death. CONCLUSION: No long-term anticoagulation in outpatients with a moderate clinical probability of PTE, a low probability V/Q lung scan and a venous lower extremities echography-doppler negative for DVT may be secure in our area. PMID- 16351490 TI - [Progressive multifocal leucoencephalopathy in AIDS patients. Hepatotoxicity of HAART in patients with hepatitis B or C]. AB - Progressive multifocal leukoencephalopathy (PML) develops in 1-8% of patients with AIDS, for which it is a disease-defining condition. PML presents mainly in severely immunocompromised male intravenous drug users, having viral loads greater than log5 RNA copies/mL and CD4 populations lower than 150 cells/mm3. Death of AIDS patients with PML occurred after only 4 to 6 months before the introduction of highly active antiretroviral therapies (HAART), the only ones that have shown to prolong survival. Viral hepatitis is not the only liver condition affecting patients with AIDS, opportunistic infections and neoplasms, such as lymphoma and Kaposi's sarcoma, as well as biliary disease are also encountered but, fortunately, they are currently less frequent as a result of the new antiretroviral treatments. The risks of HAART hepatotoxicity in patients with hepatitis B or C have been studied by several groups. PMID- 16351492 TI - [Erythema nodosum becoming a pheochromocytoma]. AB - The erythema nodosum is a septal panniculitis of several etiologies: infectious, medicamentous, secondary to systemic disease or idiopathic cause. The patient is a 47 year-female with a hypertension history of several years of evolution, who shows the following set of symptoms: successive rashes of erythematous and pruriginous cutaneous lesions in inferior members, which are diagnosed by a cutaneous biopsy of erythema nodosum. In the etiology study, an abdominal echographia is made, finding a hypodense 7 x 5 cm solid mass placed in the left adrenal gland. Determinations of catecholamines and their metabolites are found to be increased in plasma and urine, with a final diagnosis of pheochromocytoma. The pheochromocytoma is an adrenal marrow tumour of chromaffin cells derived of the neural crest, which secrete catecholamines. It is an unusual tumour, with no gender differences and with a higher incidence between the third and fourth decade of life. Ninety per cent of the cases are rather sporadic. It causes arterial hypertension just in the 0.1 and 0.2% of total cases. The appearance of erythema nodosum as one more symptom in the evolutionary context of a pheochromocytoma is a relation poorly documented in the examined literature. We could think it is just a possible coincidence, as searching with Medline we just find out one international bibliographic reference. PMID- 16351491 TI - [Meningoencephalitis by Listeria in the lupus disease]. AB - We present a patient with lupus nephropathy of 20 years of evolution in treatment with oral steroids who developed a meningoencephalitis associated to bacteraemia by Listeria monocytogenes. The patient was treated successfully with gentamicin and ampicillin for 6 weeks. Infection by Listeria monocytogenes occurs more frequently in individuals with some form of immunodeficiency like lupus disease, with a mortality around 30%. PMID- 16351493 TI - [Spontaneous tumor lysis syndrome in a patient with non-Hodgkin's lymphoma]. AB - An 77-year-old woman with no previous history of malignancy was admitted to the hospital with generalized weakness and malaise. Physical examination demonstrate multiple cervical adenopathies. The day after admission the patient developed severe metabolic derangements (hyperuricemia, hypocalcemia, hyperkalemia, hyperphosphatemia and azotemia) and expired. Consent for necropsy was refused. The cytology and molecular studies of cervical adenopathy sample were diagnostic of high-grade non-Hodgkin's lymphoma. A presumptive diagnosis of non Hodgkin lymphoma with spontaneous tumor lysis syndrome was made. PMID- 16351494 TI - [Pancreatic adenocarcinoma: therapeutical update]. AB - Cancer of the exocrine pancreas continues to be a major unsolved health problem. Because of difficulties in diagnosis, the aggressiveness of pancreatic cancers, and the lack of effective systemic therapies, generally fewer than 5% of patients with adenocarcinoma of the pancreas survive 5 years after diagnosis. Thus, incidence rates and mortality rates are virtually identical. The median survival in metastatic pancreatic cancer is nearly six months.Today, surgery remains the only curative therapeutic option, and the standard treatment in patients with advanced disease is gemcitabine. New strategies for resectable and unresectable pancreatic cancer are under active investigation,such as neoadjuvant or adjuvant chemoradiotherapy or combinations of gemcitabine with new cytotoxic agents (oxaliplatin, cetuximab, gefitinib, bevacizumab) with promising results. In patients with locally advanced pancreatic cancer and good performance status, chemoradiotherapy should be considered. PMID- 16351495 TI - [Renal failure caused by sulfadiazine in a HIV patient with toxoplasmic encephalitis]. PMID- 16351497 TI - Age-dependent relationship between the carotid intima-media thickness, baroreflex sensitivity, and the inter-beat interval in normotensive and hypertensive subjects. AB - The interrelationship between baroreflex sensitivity expressed in ms/mm Hg (BRS) or in Hz/mm Hg (BRSf), carotid wall thickness (IMT), and age was investigated in hypertensive and normotensive subjects with respect to the mean inter-beat interval (IBI) and blood pressure (BP). BP monitoring was performed in 25 treated hypertensives (Hy; 47.4+/-9.2 years of age) and 23 normotensives (Norm; 44.5+/ 8.1 years). IMT was measured by ultrasonography. BRS and BRSf were determined by the spectral method (five-minute non-invasive beat-to-beat recording of BP and IBI, Finapres, controlled breathing at a frequency of 0.33 Hz). Significant differences between Hy and Norm were detected in IMT (Hy: 0.624+/-0.183, Norm: 0.522+/-0.070 mm; p<0.01), BRS (Hy: 3.5+/-1.6, Norm: 5.7+/-2.3 ms/mm Hg; p<0.01), BRSf (Hy: 0.005+/-0.002, Norm: 0.009+/-0.004 Hz/mm Hg; p<0.01), systolic BP (Hy: 131+/-21, Norm: 116+/-17 mm Hg; p<0.01) and diastolic BP (Hy: 77+/-16, Norm: 64+/ 12 mm Hg; p<0.01). A significant correlation was found between age and IMT (Norm: 0.523, p<0.05; Hy+Norm: 0.419, p<0.01), age and BRS (Norm: -0.596, p< 0.01; Hy+Norm: -0.496, p<0.01), age and BRSf (Norm: -0.555, p<0.01; Hy: -0.540, p <0.01; Hy+Norm: -0.627, p<0.01), age and IBI (Hy: 0.478, p<0.05), age and diastolic BP (Hy: -0.454, p<0.05), BRS and IMT (Hy+Norm: -0.327, p<0.05) and BRSf and IMT (Hy+Norm: -0.358, p<0.05). Hypertensive patients have increased IMT and decreased BRS and BRSf. The positive correlation between age and IMT and the negative correlation between age and BRS and BRSf are in agreement with the hypothesis that the age-dependent decrease of baroreflex sensitivity corresponds to the age-related structural changes of the carotid wall. Using two indices of baroreflex sensitivity, BRS and BRSf, we could show that baroreflex sensitivity in hypertensives is lower not only due to thickening of the carotid wall, but also due to aging. PMID- 16351496 TI - Use of traditional, complementary and allopathic medicines in Pakistan by cancer patients. AB - INTRODUCTION: The healthcare systems of developing countries are complex in that they often accommodate a range of disparate and often competing paradigms of care. This is the case in Pakistan where Indigenous traditional medicine (TM) co exists with Western allopathic medicine and, in theory at least, with 'globalised' complementary and alternative medicines (CAM). To date we know little about what treatments are being chosen and why in this still predominantly rural country. AIM: To gain a preliminary understanding of patterns of usage of traditional medicine and globalised complementary and alternative medicine by cancer patients in Pakistan. METHOD: Structured survey of 362 cancer patients, from diverse regions in the Punjab and North-west Frontier Province provinces, who were being treated in four different hospitals in Lahore, Pakistan. RESULTS: Use of traditional medicine is high amongst cancer patients, with many patients using a combination of different therapeutic modalities. Unlike studies in Western contexts, this study indicated no relationship between cancer type or sex and use of CAM/TM. However level of education was influential in determining usage of particular TM. There is, however, no uniformity in patterns of use of different TM. CONCLUSION: There is less differentiation between social groups in usage of CAM and TM in Pakistan than has been reported in studies of western cancer patients. Differing levels of use for specific TM highlight the need to get beyond monolithic categorizations (such as TM) to understand use patterns for specific indigenous practices (in their social and cultural context). PMID- 16351498 TI - The effect of a unilateral muscle transplantation on the muscle fiber type and the MyHC isoform content in unoperated hind limb slow and fast muscles of the inbred Lewis rats. AB - To reveal the effect of foreign innervation and altered thyroid status on fiber type composition and the myosin heavy chain (MyHC) isoform expression in the rat slow soleus (SOL) and fast extensor digitorum longus (EDL) muscles, a method of heterochronous isotransplantation was developed. In this experimental procedure, the SOL or EDL muscles of young inbred Lewis rats are grafted either into the host EDL or SOL muscles of adult rats of the same strain with normal or experimentally altered thyroid status. To estimate the extent of fiber type transitions in the transplanted muscles, the SOL and EDL muscle from the unoperated leg and unoperated muscles from the operated leg could be legitimately used as controls, but only when the experimental procedure itself does not affect these muscles. To verify this assumption, we have compared the fiber type composition and the MyHC isoform content of unoperated contralateral SOL and EDL muscles and ipsilateral unoperated SOL muscle of experimental rats after unilateral isotransplantation into the host EDL muscle with corresponding muscles of the naive rats of the same age and strain. We provide compelling evidence that the unilateral heterochronous isotransplantation has no significant effect on the fiber type composition and the MyHC isoform content of unoperated muscles of experimental animals. Hence, these muscles can be used as controls in our grafting experiments. PMID- 16351499 TI - Sex differences in connexin-43 expression in left ventricles of aging rats. AB - Cardiac gap junctions have been implicated in maintaining intercellular electrical and metabolic couplings. The abnormalities in connexin-43 (Cx43) lead to conduction defects and contractile dysfunction. We have evaluated the expression and phoshorylation status of Cx43 in the left ventricular myocardium of male and female 16-month-old rats submitted to 14-week L-thyroxine (T4) treatment. Western blot analysis revealed the presence of fully or intermediately phosphorylated and unphosphorylated forms of Cx43. We have found no significant differences in Cx43 expression and phosphorylation between T4-treated and control untreated animals. However, expression of Cx43 was significantly higher in female compared to male rats. We conclude that T4 administration has no effect on Cx43 expression, but there are sex-dependent differences in Cx43 expression in the left ventricles between aging male and female rats. PMID- 16351501 TI - Fruit and vegetables consumption and gastric cancer: a systematic review and meta analysis of cohort studies. AB - Fruit and vegetable intake is widely recognized as protective for gastric cancer occurrence but prospective research challenged this belief. To evaluate the influence of design options in such results we did a meta-analysis of relevant published cohort studies identified from inception to 2004 in PubMed, EMBASE, and LILACS. Random-effects meta-analysis, stratification, and meta-regression were used to pool effects and to analyze the association with type of outcome event and length of follow-up independent of other study characteristics. An inverse association was observed between fruit intake and gastric cancer incidence (relative risk, RR = 0.82; 95% confidence interval, CI = 0.73-0.93) and stronger for follow-up periods of > or = 10 yr (RR = 0.66; 95% CI = 0.52-0.83) but not when the study outcome was death (RR = 1.08; 95% CI = 0.86-1.35). For vegetables, the RR was 0.88 (95% CI = 0.69-1.13) using all incidence studies and 0.71 (95% CI = 0.53-0.94) when considering only those with the longer follow-up. The association observed between vegetable intake and gastric cancer mortality was 1.05 (95% CI = 0.89-1.25). Other study characteristics assessed added no significant contribution to explain heterogeneity. This meta-analysis showed that design options might play a key role in the observed magnitude or the direction of the association between fruit and vegetable intake and gastric cancer. PMID- 16351502 TI - Anticancer effects of Ganoderma lucidum: a review of scientific evidence. AB - "Lingzhi" (Ganoderma lucidum), a popular medicinal mushroom, has been used in China for longevity and health promotion since ancient times. Investigations into the anticancer activity of lingzhi have been performed in both in vitro and in vivo studies, supporting its application for cancer treatment and prevention. The proposed anticancer activity of lingzhi has prompted its usage by cancer patients. It remains debatable as to whether lingzhi is a food supplement for health maintenance or actually a therapeutic "drug" for medical proposes. Thus far there has been no report of human trials using lingzhi as a direct anticancer agent, despite some evidence showing the usage of lingzhi as a potential supplement to cancer patients. Cellular immune responses and mitogenic reactivity of cancer patients have been enhanced by lingzhi, as reported in two randomized and one nonrandomized trials, and the quality of life of 65% of lung cancer patients improved in one study. The direct cytotoxic and anti-angiogenesis mechanisms of lingzhi have been established by in vitro studies; however, clinical studies should not be neglected to define the applicable dosage in vivo. At present, lingzhi is a health food supplement to support cancer patients, yet the evidence supporting the potential of direct in vivo anticancer effects should not be underestimated. Lingzhi or its products can be classified as an anticancer agent when current and more direct scientific evidence becomes available. PMID- 16351503 TI - Dietary agents in the chemoprevention of prostate cancer. AB - Nutritional factors have been estimated to contribute 20-60% of cancers around the globe, and almost one-third of deaths are being reported in Western countries. According to estimates by the American Cancer Society, during the year 2005 about 232,090 new cases of prostate cancer will be diagnosed alone in the United States and 30,350 men will die of this disease. The high incidence and long latency period of prostate cancer offer plenty of time to pursue strategies toward prevention and/or treatment to suppress or revert this disease. Epidemiological evidence suggests that plant-based dietary agents decrease the risk of some types of human cancer, including prostate cancer. Intake of 400-600 g/day of fruits and vegetables is associated with reduced risk of several cancers. The use of micronutrients and/or other phenolic agents in the diet or synthetic exogenous supplements to prevent neoplastic transformation of normal cells or to slow the progression of established malignant changes in cancer cells is termed "chemoprevention." Considerable attention has been devoted to identify plant-based dietary agents that may serve as natural inhibitors of prostate carcinogenesis. Much progress has been made in the last decade in this area of investigation through identification of pathways that play important roles in prostate tumorigenesis. This article summarizes epidemiological, clinical, and mechanistic studies and the significance of plant-derived dietary agents such as flavonoids, indoles, isothiocyanates, phenolics, monoterpenes, and complementary and alternative agents in the management of prostate cancer with recommendations for future studies to advance this area of research. PMID- 16351504 TI - Intakes of selected nutrients, foods, and phytochemicals and prostate cancer risk in western New York. AB - A number of epidemiological studies have suggested that diet may affect the etiology of prostate cancer, but few have investigated the impact of phytochemical intakes on this cancer. We conducted a case-control study of diet and prostate cancer in western New York involving 433 men with primary, histologically confirmed prostate cancer and 538 population-based controls, frequency matched to cases on age and county of residence. Diet was assessed with a detailed food-frequency questionnaire. We calculated daily intakes of nutrients and the phytochemicals beta-sitosterol, campesterol, stigmasterol, total phytosterols, total lignan precursors, quercetin, and kaempferol based on published food composition data. Odds ratios (ORs) and 95% confidence intervals (CIs) describing the association of prostate cancer risk with selected nutrients, phytochemicals, and food groups were estimated with unconditional logistic regression. Compared with men in the lowest quartile of intake, reduced risks were observed for men in the highest quartile of intake of vitamin C (OR = 0.49; 95% CI = 0.33-0.74), beta-carotene (OR = 0.53; 95% CI = 0.36-0.79), alpha carotene (OR = 0.67; 95% CI = 0.47-0.97), lutein (OR = 0.55; 95% CI = 0.37-0.81), lycopene (OR = 0.62; 95% CI = 0.42-0.92), total lignan precursors (OR = 0.66; 95% CI = 0.47-0.94), quercetin (OR = 0.64; 95% CI = 0.44-0.92), and total vegetables (OR = 0.53; 95% CI = 0.36-0.79), but weak increased risks were observed for snacks and sweets (OR = 1.46; 95% CI = 0.95-2.23). Estimates associated with nutrients and phytochemicals were attenuated after adjustment for total vegetable intake. Nevertheless, our results support the hypothesis that a phytochemical rich, plant-based diet is of importance in reducing risks of hormone-related neoplasms. PMID- 16351505 TI - Folate, vitamin B6, vitamin B12, and vitamin B2 intake, genetic polymorphisms of related enzymes, and risk of colorectal cancer in a hospital-based case-control study in Japan. AB - We conducted a case-control study to investigate the association of nutrient intake involved in the one-carbon pathway of folate for DNA methylation and DNA synthesis and the related enzyme genetic polymorphisms with colorectal cancer. Cases were 107 patients newly diagnosed with colorectal cancer. Controls were 224 subjects matched with cases by sex, age, and residential area. Nutrient intake was assessed by a self-administered, semiquantitative food-frequency questionnaire. Four genetic polymorphisms-MTHFR C677T and A1298C, MTRR A66G, and ALDH2 Glu487Lys-were determined using blood samples. Odds ratios were calculated using conditional logistic regression analysis adjusted for smoking, alcohol consumption, body mass index, and dietary fiber intake. Although folate intake was inversely associated with colorectal cancer, this association was attenuated after further controlling for dietary fiber intake. Neither vitamin B6, vitamin B12, nor vitamin B2, nor any genetic polymorphism was significantly associated with colorectal cancer. MTRR polymorphism interacted with the association of folate (P for interaction = 0.04) or vitamin (P for interaction = 0.02) with colorectal cancer, although the other polymorphisms did not interact with any nutrient intake. In conclusion, the study did not support the existing hypothesis of gene-nutrient interaction in colorectal carcinogenesis. PMID- 16351506 TI - Nutritional assessment in cancer: comparing the Mini-Nutritional Assessment (MNA) with the scored Patient-Generated Subjective Global Assessment (PGSGA). AB - The evaluation of nutritional status in cancer patients is often neglected in spite of the fact that poor nutritional status may adversely affect prognosis and treatment tolerance. In day-to-day oncology practice, a sensitive but simply applied nutritional assessment tool is needed to identify at-risk patients. Several tools exist; however, none has been universally accepted. The aim of this study was to compare two potential tools, the Mini-Nutritional Assessment (MNA) and the scored Patient Generated Subjective Global Assessment (PGSGA). The MNA is more simply applied and does not require a trained dietitian. The PGSGA has been previously validated in cancer patients. One hundred fifty-seven newly diagnosed cancer patients were assessed using both tools. Of these, 126 were reassessed at 4-6 wk, and 104 were reassessed at Weeks 8-12 after initial assessment. A significant negative correlation was found between the tools at all three time periods (at baseline r = -0.76; P < 0.001). Taking the PGSGA as the most accepted nutritional assessment tool, at baseline the MNA demonstrated a sensitivity of 97% and specificity of 54%. At 4-6 wk MNA sensitivity was 79% and specificity was 69%. At 8-12 wk MNA sensitivity was 93% and specificity was 82%. When comparing the tools in elderly patients alone (>65 yr), similar results were obtained. Both tools were able to correctly classify patients as malnourished, although the MNA lacks specificity. Therefore, the PGSGA should be the tool of choice for nutritional assessment in cancer patients. PMID- 16351507 TI - IGF-1, IGFBP-3, and nutritional factors in young black and white men: the CARDIA Male Hormone Study. AB - Nutritional factors might play a role in regulating serum levels of insulin-like growth factors (IGFs), which are associated with some cancers. We examined the associations of nutritional factors with IGF-1 and IGF binding protein-3 (IGFBP 3). Serum IGF-1 and IGFBP-3 levels and dietary intake were measured in 459 black and 682 white male subjects of the Coronary Artery Risk Development in Young Adults study at the Year 7 (1992-1993) exam. Analysis of covariance and multivariable linear regression were used to assess associations of IGFs with dietary factors by race. IGF-1 was positively associated with magnesium in both black and white men (P = 0.008 and 0.05, respectively). Calcium was positively significantly related to IGF-1 in black men (P = 0.04) and marginally so in white men (P = 0.09). In black men, IGFBP-3 was positively associated with magnesium (P = 0.02), and one serving of milk per day was associated with an 8.23-ng/ml higher IGF-1 concentration (P = 0.05). Tests for interaction, however, revealed no differences between blacks and whites in the associations of nutrients with IGF-1 or IGFBP-3. In conclusion, the associations of dietary factors with serum IGF-1 and IGFBP-3 observed in our study corroborate those from other studies and generally do not differ between black and white men. PMID- 16351508 TI - Milk consumption in relation to incidence of prostate, breast, colon, and rectal cancers: is there an independent effect? AB - Milk contains a wide variety of ingredients, such as nutrients, hormones, and chemical contaminants. Whether milk consumption is associated with the risk of prostate, breast, colon, and rectal cancers is unclear and was evaluated in this study. Data on milk consumption for 9 time periods (1964-1994) and incidence rates of prostate, female breast, colon, and rectal cancers, mostly around 1993 1997, in 38 countries were obtained from the Food and Agriculture Organization and World Health Organization, respectively. Milk consumption was strongly correlated with incidence rates of prostate cancer (r = 0.65-0.69; all P < 0.0001) and breast cancer (r = 0.64-0.74; all P < 0.0001) in all the nine time periods examined. A modest positive correlation was found for colon and rectal cancers in both sexes (all P < 0.05, except for rectal cancer in the first three time periods). The previous findings remained essentially unchanged after adjustment for vegetable, alcohol, and cigarette consumption but disappeared after further adjustment for non-milk fat consumption, except for breast cancer in the last three time periods. The present study does not support an overall substantial effect of milk consumption on the risk of prostate, breast, colon, and rectal cancers at the population level. PMID- 16351509 TI - Effect of a soymilk supplement containing isoflavones on urinary F2 isoprostane levels in premenopausal women. AB - Epidemiological studies have associated soy diets with reduced risk of breast and some other cancers, and oxidative cellular damage may contribute to the development of these and other diseases. We tested the effect of a soymilk supplement rich in isoflavones on a measure of cellular lipid peroxidation in a controlled feeding study. Eight premenopausal women consumed a constant diet that included soymilk containing 113-207 mg/day total isoflavones and 4 mo later a constant soy diet low (<5 mg/day) in isoflavones, both for a complete menstrual cycle. The average daily urinary excretions of daidzein and genistein were 24.6 +/- 10.1 and 9.2 +/- 6.1 mg/day, respectively, during the high-isoflavone soy diet and were below the detection limit during the low-isoflavone diet. F2 isoprostane 8-iso-PGF-2alpha excretion varied widely within and between subjects, and the group mean was not significantly different during the high- and low isoflavone soy diet (7.67 +/- 1.13 and 8.65 +/- 1.18 nmol/12 h, mean +/- SD, respectively). However, individual changes in 8-iso-PGF-2alpha between the two soymilk drinks were significantly associated with age (r = -0.87; P = 0.006) and several measures of isoflavone exposure, namely, daidzein dose (r = 0.81; P = 0.015), combined daidzein and genistein dose (r = 0.77; P = 0.03), and total urinary excretion of isoflavones (daidzein, genistein, and equol) (r = 0.71; P = 0.05). The findings suggest that soy isoflavones may reduce lipid peroxidation in an age-dependent manner, with greater effects in older women, and with lower doses of isoflavones. PMID- 16351510 TI - Effects of daidzein, genistein, and 17beta-estradiol on 7,12 dimethylbenz[a]anthracene-induced mutagenicity and uterine dysplasia in ovariectomized rats. AB - Phytoestrogens, primarily isoflavones daidzein (DZ) and genistein (GE), are increasingly used by postmenopausal women as an alternative to hormone replacement therapy due to reports that estrogen therapy increases the risk of breast and endometrial cancers. These compounds, as estrogen receptor agonists, may influence chemical carcinogenesis in estrogen-responsive tissues such as the uterus. We utilized ovariectomized (OVX) rats to model menopause and assessed the effects of dietary DZ, GE, or 17beta-estradiol (E2) on carcinogen-induced mutagenesis and carcinogenesis in the rat uterus. Big Blue transgenic rats (derived from Fischer 344 strain) were exposed to 7,12-dimethylbenz[a]anthracene (DMBA) in the presence or absence of the supplements. At 16- or 20-wk sacrifice, the uteri were removed and processed to determine mutant frequencies (MFs) and immunohistochemical or histopathological parameters, respectively. In rats treated with DMBA alone, a significant increase in lacI MFs (P < 0.01) in both OVX and intact (INT) rats was observed. The DMBA-induced MFs were not significantly altered by dietary DZ, GE, or E2 in both OVX and INT rats. Although dysplasia was not induced in the uterus of OVX and INT rats treated with DMBA alone, it was detected in 55% of OVX rats fed E2 alone and in 100% of OVX rats fed E2 along with DMBA exposure. Cell proliferation also was significantly higher in OVX rats fed E2 and treated with DMBA. In rats fed the isoflavones and treated with DMBA, the incidence of dysplasia was either reduced or virtually absent in both OVX and INT groups. These results indicate that a high incidence of dysplasia was associated with E2 feeding with or without DMBA treatment in the OVX rats, whereas the incidence was low in rats fed DZ or GE and treated with DMBA, suggesting a weak estrogen receptor agonist of DZ or GE in the rat uterus. The absence of dysplasia in OVX rats exposed to DMBA alone also suggests, in part, a promotional mechanism via estrogen- or isoflavone-driven cell proliferation. PMID- 16351511 TI - Betalains, phase II enzyme-inducing components from red beetroot (Beta vulgaris L.) extracts. AB - Crude aqueous and ethanolic extracts of root tissue of red (Rd) and high-pigment (HP) beet (Beta vulgaris L.) strains exhibited antioxidant and phase II enzyme inducing activities, and these extracts were fractionated using Sephadex LH-20 chromatography. These bioactivities tended to become co-enriched in early and late eluting fractions, comprising 5-25% of the material recovered from the column. Liquid chromatography-mass spectrometry (MS) was used to resolve and identify multiple betalain components in the most potent quinone reductase (QR) inducing fractions. Active fractions were found to contain vulgaxanthins I and II, and (iso)betanin, but other components remained unidentified. Two of the isolated active fractions were incorporated into rodent diets at 10-150 ppm over a 2-mo period to assess bioavailability and in vivo efficacy for phase II enzyme induction in various organs. No statistically significant effect of diet was obtained, and wide ranges of tissue enzyme levels among individual animals were observed. This lack of effect and diversity in response to diet may be related to the wide range in absorptive capacity of and/or insufficient level or enrichment of the active agents or to difficulties in assessing such activity in vivo. Subsequent to the animal studies, betanin was isolated in pure form, identified by MS analysis, and confirmed to be QR inducers in the bioassay. PMID- 16351512 TI - Anti-angiogenesis efficacy of the garlic ingredient alliin and antioxidants: role of nitric oxide and p53. AB - Alliin, a compound derived from garlic, demonstrated dose-dependent inhibition of fibroblast growth factor-2 (FGF2)-induced human endothelial cell (EC) tube formation and angiogenesis in the chick chorioallantoic membrane (CAM) model. Additionally, alliin demonstrated potent inhibition of vascular endothelial growth factor (VEGF)-induced angiogenesis in the CAM model. The antioxidant vitamins C and E significantly (P < 0.001) enhanced the inhibitory efficacy of alliin on FGF2-induced EC tube formation and angiogenesis. Alliin significantly increased (P < 0.01) nitric oxide (NO) release into the CAM fluid, which was further enhanced by vitamins C and E. The NO synthesis inhibitor nitro-L-arginine methyl ester (L-NAME) reversed the anti-angiogenesis efficacy of alliin in the CAM model. Vitamins C and E significantly enhanced the anticancer efficacy of alliin in inhibiting colon and fibrosarcoma tumor growth. Alliin significantly inhibited both FGF2 and VEGF secretion from human fibrosarcoma cells in a concentration-dependent manner. Additionally, alliin up-regulated the p53 production in FGF2-stimulated EC. These data indicated a synergistic effect of antioxidants on the anti-angiogenesis and anticancer efficacy of alliin. These data also suggest the implication of cellular NO and p53 as mediators of anti angiogenesis and anticancer effects of alliin. PMID- 16351513 TI - Rye bran diet increases epithelial cell apoptosis and decreases epithelial cell volume in TRAMP (transgenic adenocarcinoma of the mouse prostate) tumors. AB - A rye bran diet has been suggested to retard prostate cancer growth and increase tumor cell apoptosis in animal models and in men with prostate cancer. Transgenic mice designed to develop prostate cancer (transgenic adenocarcinoma of the mouse prostate, TRAMP) were therefore treated with a rye bran diet. TRAMP mice were fed a rye bran (n = 13) or a control diet (n = 15) from 4 to 24 wk of age. Tumor histology, cell proliferation, and apoptosis were accessed using stereological and immunohistochemical techniques. In the dorsolateral prostate lobe in TRAMP mice, epithelial cell apoptosis was increased by 31%, and the volume fraction of epithelial cells was reduced by 20% (P < 0.05) in the rye bran-treated group compared with the control diet group. There were, however, no significant changes detected in lobe weights or tumor grade. In conclusion, a rye bran diet increases epithelial apoptosis and reduces the epithelial cell fraction in TRAMP tumors. PMID- 16351514 TI - Effect of dried plums on colon cancer risk factors in rats. AB - Dried plums (that is, prunes) are a fruit that show promise as a food to lower colon cancer risk, based on their high content of dietary fiber and polyphenolics. In this study, we have examined the effect of diets containing dried plums on the number of colonic precancerous lesions (aberrant crypts, ACs), fecal bile acid concentration, and cecal bacterial enzyme activities related to colon cancer risk. Rats were fed one of four diets: a basal diet (a modified AIN 93G diet), a low-concentration dried plum diet (LCDP, 4.75% dried plum powder), a high-concentration dried plum diet (HCDP, 9.5% dried plum powder), or a diet matched to the carbohydrate content of the HCDP diet (CH-M) for 10 days. All animals were then administered azoxymethane (15 mg/kg, s.c., given two times, 1 wk apart) and fed their respective diets for 9 additional weeks. The number of AC foci (ACF), large ACF (>3 AC/ACF), or ACF multiplicity (AC/ACF) did not differ among the four groups. When compared with the basal diet, rats fed the LCDP diet had significantly lower concentrations of total fecal bile acids, deoxycholic acid, and hyodeoxycholic acid. Rats fed the HCDP diet had significantly lower fecal concentrations of lithocholic acid and hyodeoxycholic acid. The LCDP and HCDP diets significantly decreased the cecal activity of 7alpha-dehydroxylase, and the LCDP also had lower beta-glucuronidase activity. The LCDP, HCDP, and CH-M groups had significantly greater cecal nitroreductase activities than the basal group. There was a significant correlation between 7alpha-dehydroxylase activity and fecal lithocholic acid concentration. Compared with the basal diet, both the LCDP and HCDP diets greatly increased cecal supernatant oxygen radical absorbance capacity (ORAC). These results suggest that, although dried plums did not reduce ACF number, they favorably altered other colon cancer risk factors. PMID- 16351516 TI - Evidence for our opinions. PMID- 16351518 TI - "Virtual" pediatric palliative care: a novel concept for small pediatric programs. PMID- 16351517 TI - Expanding the tools used to promote palliative care. PMID- 16351520 TI - Infrastructure supports what is most important in palliative care. PMID- 16351521 TI - Accusations of murder and euthanasia in end-of-life care. PMID- 16351522 TI - From cure to palliation: staff communication, documentation, and transfer of patient. AB - In the transition from curative treatment to palliative care of a general end-of life patient population, the internal communication of the acute care staff seems to be less than optimal. The communication had reference to the dialogue within the staff both before and after the decision to concentrate on palliative care, and possible transfer of the patient. This survey of Swedish nurses and physicians showed that most of 780 respondents wanted more internal communication, and a more individualized procedure of decision-making. All staff should be informed about the decision made but full agreement was not seen as realistic. The largest difference of opinion between nurses and physicians concerned the involvement of nurses in the decision-making about the transition. A uniform documentation of the decision to transfer care focus was the ideal. Approximately every fourth patient in acute care is transferred to receive palliative care. Only approximately half of the respondents had any training in palliative care and the majority wanted more training. There seems to be a need for more palliative care training, perhaps somewhat different for each specialty. Furthermore, a common language to enable nurses and physicians to communicate more easily may improve the transition process. PMID- 16351523 TI - Assessment and response to spiritual pain: part II. AB - In Part I of this article, published in October 2005, I hypothesized that the components of spiritual pain can be summarized in the following manner. Spiritual pain or Suffering = ((Awareness of death + Loss of Relationships + Loss of Self) (Loss of Purpose + Loss of Control))/(Life affirming and transcending Purpose + Internal Sense of Control). Thus, an assessment of spiritual pain or suffering should examine the degree to which the individual is experiencing each of these components and their relationship to each other. Now in Part II I examine the Christian sufferer's religious responses and suggest pastoral interventions. PMID- 16351524 TI - Gabapentin is effective in the treatment of cancer-related neuropathic pain: a prospective, open-label study. AB - BACKGROUND: Gabapentin has been evaluated in the treatment of nonmalignant neuropathic pain, however, there is little direct evidence evaluating its efficacy in cancer-related neuropathic pain. OBJECTIVE: This study evaluated the effectiveness of gabapentin to treat cancer-related neuropathic pain. DESIGN: This was an open-label study. Two parallel groups of patients were recruited with either treatment-related (radiotherapy, surgery, chemotherapy) or tumor-related neuropathic pain. Gabapentin was dose-escalated from 300 mg/d to 1.8 g/d. MEASUREMENTS: The primary outcome, pain, was assessed using the modified brief pain inventory. In addition patient descriptors of pain and scores of activities of daily living were collated together with demographic data. RESULTS: We recruited 62 patients with treatment-related (n = 25) or tumor-related (n = 37) neuropathic pain. There was a significant reduction in the worst, average, and current pain scores (p < 0.002), but not the least pain score (p = 0.21). Twenty eight of 62 (45.2%) of patients achieved at least a one third reduction in pain score (95% confidence interval [CI] 32.5-58.3); the number needed to treat to obtain this benefit is 2.2 (95% CI 1.7-3.1). There was a significant reduction in all scores measuring the impact of pain on daily living (p < 0.003). There was no significant difference in pain scores at day 8 compared to day 15. Analysis of variance suggested that gender, but not etiology, or type of neuropathic pain, was a significant predictor of analgesic response and this warrants further investigation. CONCLUSION: We conclude that gabapentin is an effective treatment for cancer-related neuropathic pain. PMID- 16351525 TI - The growth of palliative care programs in United States hospitals. AB - BACKGROUND: Palliative care programs are becoming increasingly common in U.S. hospitals. OBJECTIVE: To quantify the growth of hospital based palliative care programs from 2000-2003 and identify hospital characteristics associated with the development of a palliative care program. DESIGN AND MEASUREMENTS: Data were obtained from the 2001-2004 American Hospital Association Annual Surveys which covered calendar years 2000-2003. We identified all programs that self-reported the presence of a hospital-owned palliative care program and acute medical and surgical beds. Multivariate logistic regression was used to identify characteristics significantly associated with the presence of a palliative care program in the 2003 survey data. RESULTS: Overall, the number of programs increased linearly from 632 (15% of hospitals) in 2000 to 1027 (25% of hospitals) in 2003. Significant predictors associated with an increased likelihood of having a palliative care program included greater numbers of hospital beds and critical care beds, geographic region, and being an academic medical center. Compared to notfor- profit hospitals, VA hospitals were significantly more likely to have a palliative care program and city, county or state and for-profit hospitals were significantly less likely to have a program. Hospitals operated by the Catholic Church, and hospitals that owned their own hospice program were significantly more likely to have a palliative care program than non- Catholic Church-operated hospitals and hospitals without hospice programs respectively. CONCLUSIONS: Our data suggest that although growth in palliative care programs has occurred throughout the nation's hospitals, larger hospitals, academic medical centers, not-for-profit hospitals, and VA hospitals are significantly more likely to develop a program compared to other hospitals. PMID- 16351526 TI - Tolerability of the compound ABHR in hospice patients. AB - BACKGROUND: Compounded formulations comprised of multiple medications may be useful for some patients with refractory nausea and vomiting. The combination of lorazepam, diphenhydramine, haloperidol, and metoclopramide (ABHR) has been used for more than a decade in the management of nausea and vomiting, but the tolerability of this combination in hospice patients has not been examined. OBJECTIVE: Determine the tolerability of the ABHR compound in both younger and elderly hospice patients. DESIGN: Retrospective cohort study. SETTING/SUBJECTS: A total of 11,181 ABHR prescriptions dispensed to 8600 hospice patients in the home care setting were analyzed. RESULTS: A total of 42 ABHR prescriptions were discontinued secondary to adverse drug reactions (ADRs) in 39 patients. Older patients appear to be more sensitive to the effects of ABHR, however, this trend did not achieve statistical significance when looking at extrapyramidal symptoms (EPS) specifically in these patients. The overall incidence of EPS was low, as only 0.1% of all prescriptions discontinued secondary to an ADR were consistent with EPS. CONCLUSIONS: The compound ABHR is generally well tolerated in the majority of hospice patients. More investigation is warranted to determine the efficacy and tolerability of similar pharmaceutical compounds used commonly in the hospice patient population. PMID- 16351527 TI - The association between anemia and fatigue in patients with advanced cancer receiving palliative care. AB - BACKGROUND: Fatigue has been reported to be associated with anemia in patients receiving cancer treatment. Treatment of anemia such as erythropoietin has been reported to decrease fatigue in these patients. OBJECTIVE: To investigate the correlation between anemia and fatigue intensity in patients with advanced cancer receiving palliative care. METHODS: We reviewed medical charts of 177 consecutive outpatients seen by our palliative care specialists. Information of fatigue intensity and hemoglobin level was collected. RESULTS: Among 147 (83%) evaluable patients, the median hemoglobin level was 11.6 g/dL (range, 7.5-16.1). Eighty-two (56%) patients had a hemoglobin level 12 g/dL or less, whereas 125 (85%) had 10 g/dL or more. The median fatigue score in patients with a hemoglobin level 10 g/dL or more and 10 g/dL or less was 6 (range, 4-8) and 7 (range, 5-8), respectively (p = 0.048). The median fatigue score in patients with a hemoglobin level 12 g/dL or more and 12 g/dL or less was 6 (range, 4-7) and 6 (range, 4-8), respectively (p > 0.5). Spearman's rank correlation coefficient showed a significant association only between the hemoglobin level and the albumin level (r = 0.52, p < 0.0001). Hemoglobin level did not show a significant correlation with fatigue although there was a trend (p = 0.09). In a multivariate regression analysis of the intensity of fatigue and other clinical variables, three variables remained significant in the reverse elimination analysis: depression (p = 0.0067), albumin level (p = 0.0079), and sensation of well-being (p = 0.0569). The overall explained variance for this model was 0.22. CONCLUSION: Our findings suggest that anemia is not one of the major contributors to fatigue in patients with cancer receiving palliative care. PMID- 16351528 TI - Management of oropharyngeal and tracheobronchial secretions in patients with neurologic disease. AB - BACKGROUND: Neurologic disorders may impair the normal clearance of secretions. Effective palliation requires the management of excessive oral, pharyngeal and/or tracheobronchial secretions. This requires an understanding of underlying mechanisms and familiarity with the many available medical and surgical treatment options. OBJECTIVES: The authors intend to review the relevant anatomy and physiology along with the available medical, surgical and physical therapies available to treat this commonly encountered problem. DESIGN: A review of current management and the supporting literature. CONCLUSIONS: Clinicians have many effective therapeutic options to choose from when managing the excessive oral, pharyngeal and/or tracheobronchial secretions caused by neurologic disorders. Treatment choices that are predicated upon pathophysiologic causes and patient status are the most likely to succeed. PMID- 16351529 TI - Oregon hospice chaplains' experiences with patients requesting physician-assisted suicide. AB - BACKGROUND: Oregon's Death with Dignity Act (ODDA), which legalized physician assisted suicide (PAS) for terminally ill individuals, was enacted in 1997. Eighty-six percent of the 171 patients who have died by PAS were enrolled in hospice. OBJECTIVE: To survey hospice chaplains regarding their views on the ODDA and experiences working with patients who request PAS. DESIGN: Single, anonymous, mailed survey. SUBJECTS: All chaplains affiliated with one of Oregon's 50 hospices. RESULTS: Fifty of 77 hospice chaplains whom we identified (65%) returned the survey. Forty-two percent of respondents opposed the ODDA and 40% supported it. Over half of respondents had, in the previous 3 years, worked with a patient who had made an explicit request for assisted suicide. Conversation with patients around PAS focused on the role of faith and spirituality in this decision, reasons for wanting hastened death, and family concerns or reactions to PAS. Chaplains did not feel that they had a strong influence on the patient's decisions about PAS (mean score of 4 on a 0-10 scale), though three chaplains reported a patient who withdrew their request for PAS because of the chaplain's involvement. Chaplains reported provision of a nonjudgmental presence helped the relationship with the patient. CONCLUSION: Oregon hospice chaplains are divided in their views on legalized PAS, but primarily see their role to deliver support to patients no matter what the patient's final decision regarding PAS. PMID- 16351530 TI - Men as caregivers at the end of life. AB - BACKGROUND: Few studies have focused on men as caregivers at the end-of-life. The objective of this secondary data analysis was to examine the experiences of men involved in end-of-life caregiving, focusing on caregiver strain. METHODS: We used a random sample of Oregon death certificates to telephone survey family caregivers of Oregonians who had died 2 to 5 months earlier in private homes, nursing homes, and other community-based settings. Measurements included single item indicators and embedded scales to measure caregiver strain and perceived decedent symptom distress. For the 25 husbands, sons, wives, and daughters who reported the highest levels of strain, we also analyzed caregivers' description of the decedent's last few days of life. RESULTS: The sample included 1384 caregiver interviews from a pool of 3048 death certificates. Men constituted 29% of the caregivers, including 15% sons, 9% husbands, and 5% others. In a linear regression model, male gender was a significant predictor of lower caregiver strain (p < 0.001). The strongest predictor of high end-of-life caregiver strain was the severity of the decedents' symptom distress. The qualitative analysis revealed that men used fewer words than women did to describe their experiences, and, despite subsequently reporting the highest levels of caregiving strain, only 15% of men spontaneously mentioned their own struggles. CONCLUSIONS: As caregivers at the end of life, men are less common and less likely to report caregiver strain and decedent symptom distress. Health care professionals should actively ask men about these issues and listen carefully, as their responses may be brief and understated. PMID- 16351531 TI - Communication between physicians and family caregivers about care at the end of life: when do discussions occur and what is said? AB - BACKGROUND: Few studies have examined physician-family caregiver communication at the end of life, despite the important role families have in end-of-life care decisions. We examined family caregiver reports of physician communication about incurable illness, life expectancy, and hospice; the timing of these discussions; and subsequent family understanding of these issues. DESIGN: Mixed methods study using a closed-ended survey of 206 family caregivers and open-ended, in-depth interviews with 12 additional family caregivers. SETTING/SUBJECTS: Two hundred eighteen primary family caregivers of patients with cancer enrolled with hospice between October 1999 and June 2002. MEASUREMENTS: Family caregiver reports provided at the time of hospice enrollment of physician discussions of incurable illness, life expectancy, and hospice. RESULTS: Many family caregivers reported that a physician never told them the patient's illness could not be cured (20.8%), never provided life expectancy (40% of those reportedly told illness was incurable), and never discussed using hospice (32.2%). Caregivers reported the first discussion of the illness being incurable and of hospice as a possibility occurred within 1 month of the patient's death in many cases (23.5% and 41.1%, respectively). In open-ended interviews, however, family caregivers expressed ambivalence about what they wanted to know, and their difficulty comprehending and accepting "bad news" was apparent in both qualitative and quantitative data. CONCLUSION: Our findings suggest that ineffective communication about end-of-life issues likely results from both physician's lack of discussion and family caregiver's difficulty hearing the news. Future studies should examine strategies for optimal physician-family caregiver communication about incurable illness, so that families and patients can begin the physical, emotional, and spiritual work that can lead to acceptance of the irreversible condition. PMID- 16351532 TI - Using reflection activities to enhance teaching about end-of-life care. AB - BACKGROUND: While reflection on experiences and attitudes related to death can be an important component of learning to care for the dying, opportunities within the medical school curriculum to promote learner reflection are often limited. OBJECTIVE: To describe implementation and evaluation of activities in an end-of life curriculum to promote self-reflection and student concerns and thoughts revealed through reflection exercises. DESIGN: Within a lecture-based course on end-of-life care, second-year medical students participated in four activities to promote self-reflection, including: (1) visualizing their own deaths; (2) documenting their experiences with death; (3) writing essays reacting to course content; and (4) participating in physician-led small group sessions. MEASUREMENTS: Student reaction essays and small group evaluations. RESULTS: Qualitative analysis of student reaction essays (2002-2004; total, n = 522) revealed nine major themes in student concerns and thoughts related to caring for the dying. These included: (1) expressing emotions; (2) personal grief and emotional detachment; (3) communicating effectively; (4) spending enough time; (5) feeling ill-prepared to deal with death; (6) losing a loved one; (7) shifting emphasis from curing to caring; (8) listening to patients and respecting their decisions and (9) previous negative end-of-life experiences. Analysis of essays and evaluations assessed the impact of the various reflection activities and demonstrated that these activities allowed students to critically examine and discuss their experiences and concerns regarding providing end-of-life care. CONCLUSIONS: A variety of activities can be incorporated successfully into a largely didactic curriculum to promote student reflection on their attitudes and concerns related to end-of-life care, thereby laying an important foundation for their future practice in caring for the dying. In addition, analysis of student reflection essays can serve as an important curriculum evaluation tool. PMID- 16351533 TI - Operational and financial performance of publicly traded hospice companies. AB - BACKGROUND: The performance of hospices owned by investor-owned, publicly traded hospice companies has been largely ignored by the literature. OBJECTIVE: The objective of this study was to perform a descriptive analysis that compares the operating and financial performance of hospices owned by publicly traded companies to private, for-profit hospices and to nonprofit hospices within small- and large-size categories based on patient days. DESIGN: A nonparametric median test was conducted using comparisons of median values for each measure between comparison ownership groups within the small and large size categories. SETTING: Financial and operational data for the three ownership groups included in our sample were obtained from the Centers for Medicare and Medicaid Services Cost Report Data over a 3-year period with the most recent fiscal year ending between September 30, 2002 and September 29, 2003. MEASUREMENT: We measured the operational and financial performance of hospices in three areas: utilization, services, and financial performance. RESULTS: Small hospices owned by publicly traded companies incurred a longer length of stay, lower operating expenses, generated higher revenue per day and profit margin, and served a greater proportion of Medicare patients compared to nonprofit counterparts. Large hospices owned by publicly traded hospices served a greater proportion of Medicare patients, offered fewer non-core services, had higher revenue per day and profit margin and incurred lower salary and benefit expense per day. CONCLUSIONS: Results suggest publicly traded for-profit hospices, in comparison to for-profit and nonprofit hospices, are able to earn substantially higher profits. PMID- 16351534 TI - Access to nursing home hospice: perspectives of nursing home and hospice administrators. AB - BACKGROUND AND OBJECTIVES: Hospice improves the quality of end of life care in nursing homes but serves less than 10% of dying residents. For residents to elect hospice, nursing homes must first contract for these services. We surveyed nursing home and hospice administrators to describe facilitators and barriers to hospice in nursing homes, and to test whether nursing home administrators' attitudes correlate with hospice use. METHODS: In a mailed survey, all nursing home and hospice administrators in North Carolina responded to items on hospice's effect on quality of care, and on facilitators and barriers to its use in nursing homes. Among nursing home administrators, bivariate analyses were used to test associations of attitudes with use of hospice. RESULTS: After 2 mailings, 241 (62%) nursing home administrators and 74 (85%) hospice administrators responded. Eighty-three percent of nursing homes had a hospice contract, with a median of 3 residents enrolled in the last 3 months. Nursing home administrators were less likely than hospice administrators to believe that hospice improves quality of care for pain, emotional and spiritual needs, and bereavement support. Nursing home administrators were more likely to agree that, "Nursing homes provide good care without using hospice for dying residents and their families," (24% versus 1%, p < 0.001). Among nursing home administrators with a hospice contract (n = 180), those who agreed that hospice improves quality of care had higher rates of hospice use in their facilities. CONCLUSIONS: Nursing home administrators' attitudes toward hospice may influence its availability for nursing home residents. PMID- 16351535 TI - Evaluation of a Comprehensive, Adaptable, Life- Affirming, Longitudinal (CALL) palliative care project. AB - BACKGROUND: Many persons with life-threatening illnesses require services before they begin receiving hospice or hospital-based palliative care. Although health care professionals focus on diagnosis, treatment, and comfort care, patients and families view the illness as it impacts their whole life. OBJECTIVE: Evaluating a Comprehensive, Adaptable, Life-Affirming, Longitudinal (CALL) intervention to patients with life-threatening cancer, cardiac illness, respiratory conditions or dementia. The CALL Care interventions are consistent with the 2004 Clinical Practice Guidelines for Quality Palliative Care. DESIGN: Prospective observational study. SETTING AND SUBJECTS: Eleven sites (1 primary care, 8 acute care, and 2 long term care) located around the United States. Two hundred ninety five patients and families enrolled in the study; many were enrolled for over 12 months. MEASUREMENTS: Patient self-reported Modified City of Hope Questionnaire addressing physical, psychosocial, emotional, and spiritual status and health care experiences; completed at enrollment and every three months. Family members completed Modified City of Hope Bereaved Family Questionnaire, with contents similar to the patient questionnaire. Sites compiled health care and community services utilization data regularly. RESULTS: Participants reported improved pain and symptom management. Fewer than one third were hospitalized (29%), and 7% had a critical care admission while enrolled. In addition, participants had high use of hospice (48% of those who died), and were likely to die at home (38% of those who died). CONCLUSIONS: CALL Care is an effective interdisciplinary intervention approach to individualize care for people living with life-threatening illness. PMID- 16351536 TI - A comparison of methodologies from two longitudinal community-based randomized controlled trials of similar interventions in palliative care: what worked and what did not? AB - BACKGROUND: Methodological challenges such as recruitment problems and participant burden make clinical trials in palliative care difficult. In 2001 2004, two community-based randomized controlled trials (RCTs) of case conferences in palliative care settings were independently conducted in Australia--the Queensland Case Conferences trial (QCC) and the Palliative Care Trial (PCT). DESIGN: A structured comparative study of the QCC and PCT was conducted, organized by known practical and organizational barriers to clinical trials in palliative care. RESULTS: Differences in funding dictated study designs and recruitment success; PCT had 6 times the budget of QCC. Sample size attainment. Only PCT achieved the sample size goal. QCC focused on reducing attrition through gate-keeping while PCT maximized participation through detailed recruitment strategies and planned for significant attrition. Testing sustainable interventions. QCC achieved a higher percentage of planned case conferences; the QCC strategy required minimal extra work for clinicians while PCT superimposed conferences on normal work schedules. Minimizing participant burden. Differing strategies of data collection were implemented to reduce participant burden. QCC had short survey instruments. PCT incorporated all data collection into normal clinical nursing encounters. Other. Both studies had acceptable withdrawal rates. Intention-to-treat analyses are planned. Both studies included sub-studies to validate new outcome measures. CONCLUSIONS: Health service interventions in palliative care can be studied using RCTs. Detailed comparative information of strategies, successes and challenges can inform the design of future trials. Key lessons include adequate funding, recruitment focus, sustainable interventions, and mechanisms to minimize participant burden. PMID- 16351537 TI - Palliative care services, patient abandonment, and the scope of physicians' responsibilities in end-of-life care. AB - BACKGROUND: Palliative care consultation services are being established at a growing pace in medical centers throughout the country. The intervention of these services may improve the quality of end-of-life care in many ways, but it may also promote an unintended outcome of patient abandonment by primary physicians. OBJECTIVE: To discuss the nature of patient abandonment in end-of-life care, the moral problems that it poses for palliative care clinicians in their consultative activities, and the implications of patient abandonment for palliative care services. DESIGN: Case study and conceptual analysis of two cases from the experience of the University of Pittsburgh Medical Center Palliative Care Service. CONCLUSIONS: The problem of patient abandonment raises deep questions about the proper scope of physicians' responsibilities to dying patients, and unmasks inherent tensions between the goals and functions of palliative medicine services. We offer suggestions on how palliative care services might deal effectively with these tensions, to minimize patient abandonment, and more effectively realize their moral mission. PMID- 16351538 TI - The challenge of human immunodeficiency virus: a model for palliative care. PMID- 16351539 TI - Palliative care for patient with human immunodeficiency virus/acquired immune deficiency syndrome. PMID- 16351540 TI - End-of-life and palliative care policy: social work's perspective and contributions. PMID- 16351541 TI - Promoting effective social work policy in end-of-life and palliative care. AB - Policy developments in the health care arena in general, and in end-of-life and palliative care in particular, strongly influence the practice environment for the delivery of hospice and palliative care services and the diverse roles of social work practitioners. This article analyzes policy developments in two crucial areas-ethics and law and disenfranchisement and health disparities. It focuses on the recent social work literature and other key sources and provides recommendations to promote the roles of social workers in ethics consultation, public policy, and advocacy and to integrate better the perspectives and concerns of diverse communities into palliative care practice, education, and policy. PMID- 16351542 TI - Is it pain or addiction? #68. PMID- 16351546 TI - Treatment of pruritus caused by cholestasis with opioid antagonists. PMID- 16351551 TI - Disease management and the Medicare Modernization Act: "It's the insurance, stupid". AB - While definitions of "disease management" (DM) emphasize quality of care for populations with chronic illness, proponents argue it reduces healthcare costs. Buyers may find disease management organizations' (DMOs') use of clinical guidelines, physician collaboration, and promotion of patient self-management intuitively sound, but it is performance guarantees, combined with retrospective effectiveness cost studies, that have driven DMOs' penetration of the commercial insurance market with revenues that exceed $500 million per year. The success of DMOs contributed to the creation of the Chronic Care Improvement Program (CCIP), which is designed to prospectively test the impact of DM on both the quality and cost of care for fee-for-service Medicare beneficiaries with chronic illness. This may lead to an expansion of DM in Medicare, and even greater opportunities for DMOs beyond the $10 billion in 10- year projected growth. For community-based physicians caring for patients with chronic illness, the sharpened focus on chronic care and the growth of DMOs creates some potential advantages. These include more time to treat more patients with acute illness, lower practice costs, opportunities to collaborate over quality, and a greater ability to achieve quality targets set by pay-for-performance arrangements. PMID- 16351552 TI - Asthma management programs in managed care organizations. AB - The aim of this work was to investigate how managed care organizations (MCOs) currently approach asthma treatment and management and to determine factors affecting asthma outcomes. A Web-based survey was administered to a national sample of 351 medical directors of MCOs to investigate the asthma management program components in their organizations as well as gaps and barriers in the management of patients with asthma. All 134 (38.2%) responding medical directors reported that their organizations monitor asthma patients. Plans use a variety of asthma management activities, including general member education (90%), member education by mail (87%), self-management education (85%), and provider education (82%). Educational resources (89%) and telephone advice nurse (77%) were the most common self-management strategies offered. Among factors impeding the provision of effective asthma care, noncompliance with asthma treatment, the inappropriate use of medications, and the need for multiple medications were cited by virtually all respondents. Health plans rely on an array of strategies to manage asthma patients. Education encouraging patient self-management is a key component of asthma management programs. However, a considerable number of treatment approach barriers are impeding the achievement of proper asthma care. Without innovative approaches to care, it appears that current MCOs' asthma management efforts may not result in substantial improvements in asthma outcomes. PMID- 16351553 TI - What can a pilot congestive heart failure disease management program tell us about likely return on investment?: A case study from a program offered to federal employees. AB - In 1999, the Blue Cross and Blue Shield Federal Employee Program (FEP) implemented a pilot disease management program to manage congestive heart failure (CHF) among members. The purpose of this project was to estimate the financial return on investment in the pilot CHF program, prior to a full program rollout. A cohort of 457 participants from the state of Maryland was matched to a cohort of 803 nonparticipants from a neighboring state where the CHF program was not offered. Each cohort was followed for 12 months before the program began and 12 months afterward. The outcome measures of primary interest were the differences over time in medical care expenditures paid by FEP and by all payers. Independent variables included indicators of program participation, type of heart disease, comorbidity measures, and demographics. From the perspective of the funding organization (FEP), the estimated return on investment for the pilot CHF disease management program was a savings of $1.08 in medical expenditure for every dollar spent on the program. Adding savings to other payers as well, the return on investment was a savings of $1.15 in medical expenditures per dollar spent on the program. The amount of savings depended upon CHF risk levels. The value of a pilot initiative and evaluation is that lessons for larger-scale efforts can be learned prior to full-scale rollout. PMID- 16351554 TI - Patient satisfaction, preventive services, and emergency room use among African Americans with type 2 diabetes. AB - The aim of this study was to determine the relationship between patient satisfaction and diabetes- related preventive health care and emergency room (ER) use. We studied 542 urban African-Americans with type 2 diabetes aged > or =25 years who were enrolled in a primary carebased intervention trial to improve diabetes control and reduce adverse health events; 73% female, mean age 58 years, 35% had yearly household incomes of <$7500, and all participants had health insurance. All completed a baseline interview-administered questionnaire. Patient satisfaction was measured using a modified version (nine questions) of the Consumer Assessment of Health Plans Survey (CAHPS) and use of diabetes-related preventive health care and ER were assessed by self-report. We then followed participants for 12 months to determine ER use prospectively. In general, participants gave favorable ratings of their care; over 70% reported that they had no problem getting care, over 60% reported the highest ratings on the communication and courtesy domains, and mean ratings (0-10 scale) for personal doctor and overall health care were high (8.8 and 8.4, respectively). Using poisson regression models adjusted for age, education, and self-reported rating of health, several aspects of patient satisfaction were associated with subsequent ER use. Participants who reported that medical staff were usually helpful or that doctors and nurses usually spent enough time were 0.49 and 0.37 times, respectively, less likely to use the ER (all p < 0.05). However, few aspects of patient satisfaction were associated with better preventive services. These data suggest that greater patient satisfaction was associated with lower ER use in urban African-Americans. Whether measures to improve patient satisfaction would reduce ER use requires further prospective study. PMID- 16351555 TI - Remote physiological monitoring: clinical, financial, and behavioral outcomes in a heart failure population. AB - This article reports on the outcomes associated with remote physiological monitoring (RPM) conducted as part of a heart failure disease management program. Claims data, medical records, data transmission records, and survey results for 91 individuals ages 50-92 (mean 74 years) successfully completing a heart failure RPM program were analyzed for time periods before, during, and after the monitoring intervention. The program was associated with significant reductions in per member per month costs and emergency room and hospital utilization. More detailed analyses were performed for specific gender and age subgroups. Participant surveys indicated high levels of satisfaction, and improvements in self-perceived health status, self-efficacy, and self-management behaviors. This study is the first to assess the impact of a RPM program following removal of the monitoring equipment. The results indicate that RPM, as a component of a traditional disease management program, has a sustained, beneficial effect on participants' lifestyles after the monitoring period has ended. PMID- 16351556 TI - Impact of a migraine management program on improving health outcomes. AB - The goals of this program were to evaluate the effectiveness of migraine disease management techniques in improving patient satisfaction with migraine care, decreasing the frequency and severity of headaches and migraine-associated disability, increasing the effectiveness of migraine treatment and work productivity, improving physician diagnosis and treatment of migraine, and ultimately determining the program's impact on healthcare resource utilization. A prospective observational study was undertaken. This prospective Migraine Management Program (MMP) used active patient and healthcare physician-based disease management resources, tools, and techniques. Members were identified using administrative and medical claims databases indicating an ICD-9 diagnosis code (346.XX) for migraine during the previous twelve months. All identified patients received the Migraine Therapy Assessment Questionnaire (MTAQ) to assess their level of migraine control for pre/post measurement. Of the 2,134 patients responding to the initial MTAQ, 789 completed both a baseline and follow-up and 1,345 completed only a baseline questionnaire. For those patients completing both, there was a statistically significant reduction in all identified management issues: poor symptom control, high attack frequency, knowledge/behavior barriers, economic burden, and dissatisfaction with treatment. Comparing the former group to those completing only the baseline MTAQ, the latter were significantly more likely to report problems with three migraine management issues; poorer symptom control, greater economic burden, and dissatisfaction with their treatment. The use of appropriate patient and physician educational interventions, such as Aetna's MMP incorporating disease management principles and the MTAQ questionnaire, can have a significant impact on patient-centered outcomes and satisfaction with their migraine treatment. PMID- 16351557 TI - DMAA quality and research initiative. PMID- 16351559 TI - Dexamethasone: by safe means, by fair means. PMID- 16351563 TI - L-arginine supplementation enhances exhaled NO, breath condensate VEGF, and headache at 4,342 m. AB - We examined the effect of dietary supplementation with L-arginine on breath condensate VEGF, exhaled nitric oxide (NO), plasma erythropoietin, symptoms of acute mountain sickness, and respiratory related sensations at 4,342 m through the course of 24 h in seven healthy male subjects. Serum L-arginine levels increased in treated subjects at time 0, 8, and 24 h compared with placebo, indicating the effectiveness of our treatment. L-arginine had no significant effect on overall Lake Louise scores compared with placebo. However, there was a significant increase in headache within the L-arginine treatment group at 12 h compared with time 0, a change not seen in the placebo condition between these two time points. There was a trend (p = 0.087) toward greater exhaled NO and significant increases in breath condensate VEGF with L-arginine treatment, but no L-arginine effect on serum EPO. These results suggest that L-arginine supplementation increases HIF-1 stabilization in the lung, possibly through a NO dependent pathway. In total, our observations indicate that L-arginine supplementation is not beneficial in the prophylactic treatment of AMS. PMID- 16351564 TI - Acclimatization to oxidative stress at high altitude. AB - Hypoxia-mediated oxidative stress has been implicated in the pathophysiology of high altitude maladaptations. To explore whether prolonged exposure to high altitude can trigger an adaptive response to oxidative stress and restore redox homeostasis in the body, the study was conducted to evaluate biochemical variables related to oxidative stress and antioxidant status in humans at sea level (190 m) and following 3- and 13- month sojourns at altitude (4,500 m). After 3 months at altitude, whole-blood thiobarbituric acid reactive substances (TBARS) were significantly higher (65.6%), nonenzymatic antioxidants like ascorbic acid and caeruloplasmin were significantly lower (41% and 22%, respectively) and plasma total antioxidant status (TAS), glutathione levels, and superoxide dismutase activity were marginally altered as compared to their basal values. After 13 months at altitude, TBARS levels regressed back to preexposure levels. Plasma total antioxidant status (TAS) improved by 21%, glutathione levels by 32.8%, and plasma bilirubin by 35.8% as compared to sea level. Average concentrations of ascorbic acid and caeruloplasmin were 18% and 37% higher as compared to the subjects studied after a 3-month stay at high altitude. In addition, there was a progressive rise in erythrocytic superoxide dismutase activity and persistent hyperurecemia. The study observed that on prolonged exposure to high altitude humans could mount an effective adaptive response to oxidative stress by activating the antioxidant defense. Hence, strengthening the antioxidant defense could be an effective strategy to prevent free-radical mediated pathophysiological alterations and quicken acclimatization to oxidative stress. PMID- 16351565 TI - Neutral endopeptidase null mice are less susceptible to high altitude-induced pulmonary vascular leak. AB - Hypoxia increases pulmonary vascular leak, which is regulated in part by neutral endopeptidase (NEP). NEP is a cell-surface metalloprotease that degrades several vasoactive peptides, including endothelin-1 (ET-1) and atrial natriuretic peptide (ANP). We therefore hypothesized that NEP attenuates high altitude-induced pulmonary vascular leak. Wild-type and NEP null mice were exposed to a simulated high altitude (HA) of 6,728 m (22,000 ft; P(B) = 328 mmHg) or remained at the relatively low altitude (LA) of 1,500 m (4,920 ft; P(B) = 640 mmHg) for 24 h. Plasma ANP and ET-1 concentrations, right ventricular pressure (P(RV)), and indexes of lung injury were recorded. At HA, lung wet weight-to-body weight increased in all animals, but was greatest in the NEP wild-type mice. Vascular leak, as measured by Evans blue dye, increased only in the NEP wild-type mice at HA. P(RV) increased in both genotypes at HA. Plasma ANP concentrations increased at HA in both genotypes, but plasma ET-1 concentrations were elevated only in the NEP null mice at HA. Correlations between lung wet weight-to-body weight versus P(RV) (r = 0.56; p = 0.0136) and ANP versus P(RV) (r = -0.54; p = 0.02) were noted. We conclude that NEP null mice exposed to HA have a greater rise in ANP versus ET-1 plasma concentration, decreased pulmonary vascular pressure, and reduced high altitude-induced pulmonary vascular leak. PMID- 16351566 TI - Red cell membrane lipid changes at 3,500 m and on return to sea level. AB - Previous studies have shown that acute hypobaric hypoxia, obtained in a hypobaric chamber, and subsequent reoxygenation, give rise to modifications of the erythrocyte membrane lipid dynamics, resulting in an increased lateral diffusivity of the membrane lipids, and this was interpreted as the result of a modified lipid-protein interaction. The aim of the present study was to determine the effect of the reoxygenation condition in individuals after 3 days at an altitude of 3,500 m above sea level. Reoxygenation was a consequence of returning to sea level. Resting blood samples from both conditions were obtained, and erythrocytes were separated and immediately lysed for membrane isolation. We measured the bilayer polarity in membranes with Laurdan, a fluorescent probe. We also measured malondialdehyde in membrane lipids, an indicator of oxidative damage. We found a 12% (p = 0.016, n = 7) increase in the polarity of the membrane bilayer surface, and an increase of 70% (p = 0.005, n = 7) in the formation of malondialdehyde in the membrane after the reoxygenation condition. The membrane bilayer polarity increase is due to an oxidative modification of the phospholipid backbone after reoxygenation. People working and/or recreating at moderate altitude (3,500 m) may be at risk of erythrocyte membrane oxidative damage upon returning to sea level, and therefore a better understanding of the processes occurring upon reoxygenation may lead to proposed strategies to minimize this effect. PMID- 16351567 TI - High altitude and the eye: a case controlled study in clinical ocular anthropometry of changes in the eye. AB - Tribal natives of two spatially separate districts of Lahaul-Spiti (mean altitude = 3300 m) and Kinnaur (mean altitude = 1,700 m) in the Himalayan state of Himachal Pradesh have lived for centuries as closed isolated groups, marrying within the local community up to the recent past. Studies on highland natives in the Andes have shown differences in chest diameters, lung volumes, hematocrit, and the like, in those living at high altitudes. We wanted to study whether stay at high altitude for generations confers any change in the ocular anthropometry. No study of this nature is available in the literature. We studied the ocular parameters using Snellen's charts, Standard Royal-Air-Force Rule, A-2,500 Sonomed A-Scan, Goldmann's aplanation tonometer, and S-7,000 Autorefractometer on 50 healthy volunteers at the Indira Gandhi Medical College Out Patient Department at Shimla (mean altitude = 2,100 m). Those living at higher altitudes in Lahaul Spiti [LS] (n = 10) had significantly wider nose bridge width (p < 0.05); inter inner canthal distance [IICD] (p < 0.05); inter-outer canthal distance [IOCD] (p < 0.05); and interpupillary distance [IPD] (p < 0.05) than the group living at Kinnaur (n = 40) at lower elevation. The LS group also had narrower palpebral fissure length in the vertical dimension (p = 0.05) and a lower IOP (p = 0.002) than the Kinnaur group. The axial length, lens thickness, and anterior chamber depth were comparable in the two groups. It appears that the eye and its adnexa respond to hypoxia, ultraviolet radiations, and persistent snow cover at high altitude by altering its anthropometry in a subtle but discernible manner. This was a hitherto unexplored area in the literature. Further studies to elucidate and substantiate the findings of the study are indicated. PMID- 16351568 TI - The Rahn-Otis discovery of hypoxic ventilatory decline. PMID- 16351569 TI - The Khumbu cure. PMID- 16351570 TI - Simulating moderate altitude using normobaric hypoxia with commercially available hypoxic gas generators. PMID- 16351572 TI - Comparing calpain- and caspase-3-mediated degradation patterns in traumatic brain injury by differential proteome analysis. AB - A major theme of TBI (traumatic brain injury) pathology is the over-activation of multiple proteases. We have previously shown that calpain-1 and -2, and caspase-3 simultaneously produced alphaII-spectrin BDPs (breakdown products) following TBI. In the present study, we attempted to identify a comprehensive set of protease substrates (degradome) for calpains and caspase-3. We further hypothesized that the TBI differential proteome is likely to overlap significantly with the calpain and caspase-3-degradomes. Using a novel HTPI (high throughput immunoblotting) approach and 1000 monoclonal antibodies (PowerBlottrade mark), we compared rat hippocampal lysates from 4 treatment groups: (i) naive, (ii) TBI (48 h after controlled cortical impact), (iii) in vitro calpain-2 digestion and (iv) in vitro caspase-3 digestion. In total, we identified 54 and 38 proteins that were vulnerable to calpain-2 and caspase-3 proteolysis respectively. In addition, the expression of 48 proteins was down-regulated following TBI, whereas that of only 9 was up-regulated. Among the proteins down-regulated in TBI, 42 of them overlapped with the calpain-2 and/or caspase-3 degradomes, suggesting that they might be proteolytic targets after TBI. We further confirmed several novel TBI linked proteolytic substrates, including betaII-spectrin, striatin, synaptotagmin 1, synaptojanin-1 and NSF (N-ethylmaleimide-sensitive fusion protein) by traditional immunoblotting. In summary, we demonstrated that HTPI is a novel and powerful method for studying proteolytic pathways in vivo and in vitro. PMID- 16351573 TI - Phosphorylation of Ser158 regulates inflammatory redox-dependent hepatocyte nuclear factor-4alpha transcriptional activity. AB - In IL-1beta (interleukin 1beta)-stimulated rat hepatocytes exposed to superoxide, we have previously identified an IRX (inflammatory redox)-sensitive DR1 [direct repeat of RG(G/T)TCA with one base spacing] cis-acting activator element (nt 1327 to -1315) in the iNOS (inducible nitric oxide synthase) promoter: AGGTCAGGGGACA. The corresponding transcription factor was identified to be HNF4alpha (hepatocyte nuclear factor-4alpha). HNF4alpha DNA binding activity and transactivation potential are tightly regulated by its state of phosphorylation. However, the functional consequences of IRX-mediated post-translational phosphorylation of HNF4alpha have not been well characterized. In the setting of IL-1beta+H2O2, HNF4alpha functional activity is associated with a unique serine/threonine phosphorylation pattern. This indicates that an IRX-sensitive serine/threonine kinase pathway targets HNF4alpha to augment hepatocyte iNOS transcription. In the present study, following identification of phosphorylated residues in HNF4alpha, serial mutations were performed to render the target residues phosphorylation-resistant. Electrophoretic mobility-shift assays and transient transfection studies utilizing the iNOS promoter showed that the S158A mutation ablates IRX-mediated HNF4alpha DNA binding and transactivation. Gain-of function mutation with the S158D phosphomimetic HNF4alpha vector supports a critical role for Ser158 phosphorylation. In vitro phosphorylation and kinase inhibitor studies implicate p38 kinase activity. Our results indicate that p38 kinase-mediated Ser158 phosphorylation is essential for augmentation of the DNA binding and transactivation potential of HNF4alpha in the presence of IL 1beta+H2O2. This pathway results in enhanced iNOS expression in hepatocytes exposed to pro-inflammatory cytokines and oxidative stress. PMID- 16351575 TI - Brachial plexus treatment. AB - Brachial plexus injuries are devastating injuries that affect primarily young healthy males. For the total plexus injury, current surgical treatments have failed to achieve normal restoration of limb function but some practical goals are obtainable. This review article summarizes existing logic and approach for managing these catastrophic injuries. PMID- 16351576 TI - Updates in flexor tendon repair at zone ii. AB - This article reviews the history and current management concepts of flexor tendon lacerations. Classic and contemporary repair techniques are discussed. The most popular rehabilitation protocols are also reviewed. PMID- 16351577 TI - In vitro modeling of the influence of FVIII activity and heparin induced prolongation of APTT. AB - Anticoagulant therapy is most commonly assessed by measuring the effect of the drug on global clotting assay, such as APTT. It is known that response of the APTT to heparin may be decreased in patients with high levels of factor VIII. In this work, we have attempted to determine in vitro conditions of experiment for obtaining relationship between different concentrations of heparin and values of APTT, and to investigate influence of factor VIII on correlation between concentrations of heparin and APTT. Measurement of the effect of heparin, added in vitro in normal coagulation control plasma (NCCP) showed that heparin in concentrations from 0.1 to 1.0 IU/mL prolonged APTT from 0.73 s to 99.26 s. Linearity of the relation of natural logarithm of APTT and concentration of added heparin in plasma for concentrations from 0.5 to 1.0 IU/mL (r = 0.995), and other characteristics of the validated method (RSD = 1.17%), made possible investigation of the influence of factor VIII addition in the solution. The addition of the Factor VIII concentrate, markedly influenced these APTT results. Increased factor VIII activity shortened the APTT, having more pronounced effect in the presence of the large amounts of heparin. Increased factor VIII was associated with downward shift in the concentration -- logAPTT response curve (y = 24644 x + 30.17 vs. y = 10.864 x + 27.256). This finding suggests the possibility for modeling of ex vivo establishment of correlation between plasma activity of FVIII and needed doses of heparin for appropriate management of heparin therapy. PMID- 16351578 TI - Angiography analysis of variations of the posterior segment of the circle of willis. AB - Cerebral-vascular diseases present one of the leading problems of the modern mankind. They are followed by the risk of high mortality rate, and as such cause high level of disability with people who survive cerebral-vascular incident (stroke, apoplexy). Researches done so far proved that beginning, course and result of the cerebral-vascular diseases depend immensely of the possibility to establish collateral blood circulation and first of all on so called tertian level that is actually the circle of Willis. The circle of Willis, thanks to communicating segments, provide detour way to procure parts of the brain which, due to insufficiency, do not get enough quantity of blood. In this particular study by the analysis of 150 MRI patient's angiographies of the circle of Willis that had been processed at the Radiology Institute of the Clinic Center of University in Sarajevo, we tried to present the most common variations of the posterior segment of the circle of Willis with patients who did not have signs of the cerebral-vascular diseases. The analysis included two target groups (above 60 years old and younger than 34 years old) and both genders. By the analysis of the angiographies of the circle of Willis we reached following results: complete posterior configuration of the circle of Willis has been found with all patients in 54% of cases, but in some slightly higher percentage complete posterior configuration is noted with younger category of patients compared to elderly patients, and in some higher percentage with female compared to male patients. Out of variations that damage the posterior segment of the circle of Willis the first one, according to the frequency of occurring, is the variation of the type of unilateral fetal sort of posterior cerebral artery, and then the variation marked as unilateral aplasia or hypoplasia of the posterior communicating artery. PMID- 16351579 TI - The relationship between suicidal thoughts and psychoactive substances. AB - In this study is represented the relationship between suicidal thoughts and psychoactive abuse. In study were involved 202 students of Banjaluka's secondary schools. By questioned for risky behaviour of adolescents was found 28.7% of them have suicidal thoughts, and 144 haven't suicidal thoughts. Students from both groups use alcohol but there are not statistically significant differences among groups. The representative sample was 202 students from Banjaluka's secondary schools. The results showed that 28.7% of them have suicidal ideas. From examined students 20.2% used cannabis, and 4% other psychoactive substances, 56.9% alcohol and 35.6% smoke cigarettes. In the group with suicidal ideas, use of psychoactive substances is 8.6%, alcohol use in 63.7% cases and cannabis 36.2%, and smoking cigarettes in 48.2% cases. We may conclude that misuse of psychoactive substances is more present in group with suicidal ideas, except in case of alcohol, which was also high in group with suicidal ideas, but without statistical significances. It is also possible to conclude that there is important connection between use of psychoactive substances and presence of suicidal ideas as first step toward a suicide in adolescents. PMID- 16351580 TI - Heart murmur and anaemia in the pediatric population. AB - Innocent heart murmurs are hearth murmurs that occur in patients with a normal heart structure. They do not represent a disease of the heart and vascular system, and should not be treated as such. Iron-deficiency anaemia often causes, along with other symptoms, systolic heart murmurs and tachycardia. It appears in children of all ages representing a most common haematological paediatric disease. To establish the influence of iron-deficiency anaemia on genesis of innocent murmurs and to compare auscultatory and phono-electrocardiographic findings in patients with anaemia and heart murmurs before and after iron therapy. The study includes 120 patients with innocent heart murmurs that have been auscultated at cardio-rheumatic outpatient department of Pediatric Clinic of the Clinical Center of the University of Sarajevo, during the period from 01/01/2004 to 31/12/2004. Further diagnostic procedure, i.e. laboratory tests, diagnosed iron-deficiency anaemia in 30/120 patients. These patients have been followed in this study. 22/30 patients had systolic murmur I/II intensity of Levin scale; 8/30 patients had II/VI systolic murmurs of intensity by Levin. The highest number of examinees had 0-1 years of age, and in this group the number of boys was higher than the number of girls (M: F = 12:4). During the auscultatory and phono-ECG examinations of murmurs, 6 patients had haemoglobin values less than 95 g/l, which corresponds to an average and severe type of anaemia. 24 patients had haemoglobin values between 95 and 110 g/l, which corresponds to benign type of anaemia. The most numerous were patients aged between 0 - 1 year (3 patients with hemoglobin value Hb < 95 g/l, 13 patients with hemoglobin value Hb 95-110 g/l). All patients were treated with iron medicaments. After three months, clinical and laboratory re-evaluation was performed and it has demonstrated that after iron therapy 24 patients had level of a haemoglobin Hb >110 g/l and 6 patients had haemoglobin levels between 95 and 110 g/l. By auscultatory and phono-ECG examinations, murmurs of a level of intensity I/II was registered in only one child, while in the other 29 patients there were not any registered heart murmurs. Diagnosis of anaemia in the paediatric population group delays definitive diagnosis of heart murmurs. Innocent murmurs in children with sideropenic anaemia occur as its consequence. After adequately conducted iron therapy, i.e. cured anaemia, heart murmurs were not auscultated. PMID- 16351581 TI - Serotesting of human brucellosis on wider area of Bosnia and Herzegovina. AB - The study involved 286 individuals from different regions of Bosnia and Herzegovina, whose sera were tested in the Laboratory for specific diagnosis of human brucellosis in Microbiology Department of Medical Faculty of University in Sarajevo, during the period from 2000. to 2003. Sera were tested using Brucelloslide Test, qualitative agglutination test Rose Bengal. Using the agglutination test, we serologically confirmed a diagnosis of human brucellosis in 59 (20.62%) seropositive individuals, whereof 38 (64.40%) men and 21 (35.60%) women. Individuals with human brucellosis were the most present in the age group of 31-40 (22.03%) and 41-50 (22.03%). One serologically confirmed death case was registered. The most seropositive individuals were from Zenica-Doboj Canton (32.20%), Sarajevo Canton (28.82%), Herzegovina-Neretva Canton (23.73%), Central Bosnia Canton (13.55%) and Una-Sana Canton (1.70%). During our four-year study, it was serologically confirmed that human brucellosis is present in Bosnia and Herzegovina and, through seropositive testing, we revealed the level of general exposition to Brucella spp. on wider area of Bosnia and Herzegovina. PMID- 16351582 TI - Long term protection after hepatitis B vaccination. AB - A survey of persistence of anti-HBs after hepatitis B vaccination has shown that five years after vaccination on a sample of 152 persons, or 82.53%, stands at >10 IU/I. Long term immunogenicity of vaccinated children remained at 88.89%, health workers 79.41% and drug addicts 64.28%. The results of these studies in Bosnia and Herzegovina show the high level of protection hepatitis B vaccine against HBV infection. Vaccination against viral hepatitis B results in immunologic memory response among the vaccinated, and even after a decrease of anti-HB level following the third vaccine dose inoculation, a booster dose is not needed. Immunity remains steady and a booster dose is not recommended. PMID- 16351583 TI - Cytogenetics findings at Turner Syndrome and their correlation with clinical findings. AB - Turner Syndrome is a genetic condition in females that results from an abnormal chromosome. One of the X chromosomes is missing or misshapen in the most cells of the body. Three classics clinical symptoms of the syndrome are: incomplete sexual maturation, short stature and pterygium colli. Turner Syndrome is diagnosed by karyotyping. In the retrospective study for a twelve years period (1991-2002) correlation between clinical and cytogenetics findings was established in our Center among 47 examinees from all parts of Federation of Bosnia and Herzegovina, who had suspect clinical diagnosis of Turner Syndrome. The syndrome was demonstrated by cytogenetics examinations in 30(63,8%) examinees and excluded in 17 (36,2%) examinees. The most frequent karyotype is monosomy of X chromosome (45,X) found at 63,3%, than isochromosome of Xq (46,XisoXq) found at 16,7%, mosaic form (46,XX/45,X) and deletion of Xp (46,XdelXp) both at 6,7%, than deletion of Xq (46,XdelXq) and ring of Xp (46,XX/46,XringXp) both at 3,3%. Our results suggest that promptly and exactly diagnosis of Turner syndrome is very important due to introducing growth hormone therapy and estrogen therapy at a very young age. PMID- 16351584 TI - Serum and tissue angiotensin converting enzyme in patients with lichen planus. AB - Serum and tissue angiotensin-converting enzyme (ACE) was measured in 20 patients with lichen planus before and after therapy, and in 20 healthy individuals. Serum and tissue ACE activity was determined by spectrophotometric method using hippuryl-l-histidyl-l-leucine as a substrate. The enzyme activity is expressed in the following units: 1 U corresponds to 1 nmol of hippuric acid released by hydrolysis of hippuryl-l-histidyl-l-leucine per minute and one liter of serum or 50 mg tissue. Before therapy, serum ACE activity was significantly increased in patients with lichen planus (35.9 +/- 2.33 U/L) in comparison to healthy individuals (28.16 +/- 1.7 U/L). Tissue ACE activity was increased in patients with lichen planus (2.24 +/- 0.41 U/50 mg) in comparison to healthy individuals (1.86 +/- 0.16 U/50 mg), but the difference was not significant. After therapy, serum and tissue ACE activity decreased and no significant difference in ACE activity was found. The determination of serum ACE activity may be a good non specific parameter for the assessment of therapeutic effects. PMID- 16351585 TI - Oral acute toxicity of polyenylphosphatidylcholine (PPC) in rats. AB - Endogen phospholipids play a major role in determining the structure and nature of cell membranes. A deficiency of phospholipids in cellular membranes makes it almost impossible for the cell membrane to perform its function as a selective barrier between what passes in and out of the cell. Polyenylphosphatidylcholine chemical structure corresponds to that of endogen phospholipids, but it possesses functional superiority because of its content of unsaturated fatty acids. Polyenylphosphatidylcholine integrates in the cell membrane and organelle systems while becoming their constitutive elements. A healthy cell membrane leads to healthy cells and then healthy tissue and then to healthy organs or body systems and finally, healthy bodies and minds. For a long time, polyenylphosphatidylcholine in combination with vitamins has been used in the treatment of numerous health problems such as liver diseases, dyslipoproteinaemias and different intoxications with consequent liver failure. The main aim of toxicology studies is evaluation of the toxic potential and risks of human exposition to the substance. According to the Organization for Economic Cooperation and Development (OECD) acute oral toxicity refers to those adverse effects occurring following oral administration of a single dose of a substance or multiple doses given within 24 hours. LD50 (median lethal dose), oral, is a statistically derived single dose of a substance that can be expected to cause death in 50 per cent of animals when administered by the oral route. Our acute toxicity study was performed on albino Wistar rats. Animals were randomised in three experimental and one control group, each of 5 males and 5 females. Study was based on the administration of a single oral dose of the test substance (polyenylphosphatidylcholine) to each experimental animal. There were three dose levels of the test substance: 300, 500 and 1000 mg/kg. Test substance administration day was the first day of the observation period that lasted 14 days. Control animals were given milk vehicle. At the end of the study, no statistically significant differences between experimental and control animals were observed concerning the recorded parameters: body weight, respiratory rate, tremor, faeces and phonation quality, indicating the absence of the test substance acute toxicity. PMID- 16351586 TI - Branching of main trunk of left coronary artery and importance of her diagonal branch in cases of coronary insufficiency. AB - Proficiency in the anatomy of coronary arteries and their variations is significant for proper interpretation of the coronary angiographies, assessment of the complexity and result of the coronary insufficiency as well as surgical myocardium revascularization. The objective of this study is anatomy-radiology research of the methods of branching the main trunk of left coronary artery and to prove importance of the diagonal branch (ramus diagonalis) existence in the conditions of coronary insufficiency. In this study we have analyzed 100 coronary angiographies done at the Clinic for Heart Diseases and Rheumatism of the Clinic Center of University of Sarajevo and dissected 20 human hearts from the Institute of Anatomy. In our study we have come upon two methods of branching of main trunk of left coronary artery (bifurcation and trifurcation). By the method of the angiography we have found the bifurcation in 71% of cases while 65% of cases were proved by the dissection method. Trifurcation has been discovered in 29% of cases of analyzed angiographies i.e. 35% of cases of dissected hearts. We believe that third terminal branch of the left coronary artery should be marked as ramus diagonalis. This branch, including its anastomoses, presents important pattern of the collateral blood flow, which has special meaning, under conditions of coronary insufficiency. PMID- 16351587 TI - Modified pair technique for treatment of hydatid cysts in the spleen. AB - The aim of this study was to evaluate the results of single-session sclerotherapy with mixture of alcohol and polidocanol and a subsequent injection of albendazole for devisceration of hydatid cysts in the spleen. Eight patients (four women and four men, average age 22.9+/-11.4 with hydatid cyst in the spleen were treated with 10 minutes time of exposure to mixture of ethanol 95% and polidocanol 1%. After that, 2 to 5 ml of albendazole was injected into the cyst cavity. Two patients had 2 cysts. At follow-up the patients were examined with clinical and biochemical examinations, ultrasonography, and serologic test for echinococcal antibody titres. The mean hospital stay was 2.5+/-0.93 days. During the follow-up period, mean cyst diameter decreased from 46+/-16.4 mm to 13.6+/-16.26 mm. In all ten cysts, a reduction of post procedural recolection of fluid over 40% was observed. Five cysts (50%) disappeared during the follow-up period. All cysts (5) smaller then 50 mm in diameter disappeared during follow-up period. After an initial rise, the echinococcal-antibody titres fell progressively and at the last follow-up were negative (< 1: 160) in 7 (88%) patients. No complications were observed, except for pain, fever and urticaria during the first 24-hours after the procedure. Sclerotherapy using only one session and 10 min time of exposure to the mixture of ethanol and polidocanol, and a subsequent injection of albendasole solution represents an effective treatment of hydatid cysts in the spleen. This procedure is even more efficacious for hydatid cyst with diametar smaller then 50 mm. PMID- 16351588 TI - Detection of neurovascular structures using injection pressure in blockade of brachial plexus in rat. AB - In the last few decades there has been a great development of regional anesthesia; all the postulates are defined and all the techniques of usage are perfected. However, like any other medical procedure, the block of brachial plexus carries a risk of certain unwanted complications, like possible intraneural and intravascular injections. The reason for great discrepancy between the injury of brachial plexus and other periphery nerves while performing the nerve blockade is the frequent usage of this block, but also the specific proximity of neurovascular structures in axilla. The purpose of this work is to determine the values of pressures which appear in para-neural, intraneural and intravascular injection applications of local anesthetic, and to compare those values in order to avoid cases of intraneural and intravascular injections in clinical practice with consequential complications. In experimental study there have been used 12 Wistar rats of both genders. After anesthesia with ether and mid-humoral access to the neurovascular structures in axilla, the injection of 2% lidocaine with epinephrine was performed with the help of automatic syringe charge. The needle was at first placed para-neural, and then also intraneural and intravascular. During every application the pressure values were monitored using the manometer, and then they were analyzed by special software program. All para neural injections resulted with the pressure between 13,96-27,92 kPa. The majority of intraneural injections were combined with the injection pressure greater than 69,8 kPa, while the intravascular injections were combined with injection pressure less than 6,98 kPa. Based on the available data it can be noticed that so far none of the methods of prevention from unwanted complications of regional anesthesia can insure the avoidance of intraneural and intravascular injection of local anesthetic. Based on our research it is obvious that the measuring of pressure during the nerve blockade is very important in order to decrease the risk of neurological and possible systematic complications. It is also clear that a small, mobile, and financially quite available apparatus for pressure measurement can help in differentiation between para-neural, intraneural and intravascular injection. Avoiding high injection pressure prevents from lodging the needle into intraneural space, while avoiding a very low injection pressure prevents from lodging the needle into intravascular space followed by consequential complications. The usage of this apparatus can find its application in other blockades of periphery nerves, and in other branches of medicine as well. PMID- 16351590 TI - Editorial. PMID- 16351589 TI - Significantly reduced salivary nitric oxide synthesis in patients with Parkinson's disease. AB - In order to study concentration of nitric oxide (NO) in the saliva of patients with Parkinson's disease (PD), we measured the concentration of its stable metabolite nitrite (NO(2)-) in the saliva of these patients and healthy subjects. We analyzed saliva flow rate and salivary NO concentrations in 16 subjects with Parkinson's disease and in 16 healthy subjects. Concentration of nitrite was determined by colorimetric method using Griess reaction. Saliva flow rate was significantly lower in patients with Parkinson's disease (0.2+/-0.03 mL/min; X+/ SEM) than in healthy subjects. Salivary NO(2)-concentration was significantly lower (5.02+/-0.64) than in healthy individuals (22.39+/-1.24, p<0.0001). PMID- 16351591 TI - Timing of management of severe injuries of the lower extremity by free flap transfer. AB - Covering defects by free-tissue transfers enable surgeons to reconstruct or salvage the lower extremity injured or amputated in high-energy traumas which result in extensive damage to soft tissue, bone, tendons, vessels and nerve. The timing of the reconstruction using flap techniques is extremely important. It can be divided into three categories: "primary free flap closure" (12 to 24 hours), "delayed primary free flap closure" (2 to 7 days), and "secondary free flap closure" (after 7 days). Our treatment of choice in an isolated complex injury of a lower extremity with a soft tissue defect is "primary free flap closure" providing improved funcional and aesthetic results, and psychologically benefit through lowered morbidity of the patient. PMID- 16351592 TI - Management of open fractures. AB - The large spectrum of open fractures is an amalgamation of injuries with the single variable in common of communication of the fractured bone with the outside environment, and thus an increased risk for infection. Contributing to the presence of bacteria within the fracture site is devascularized soft tissue, the degree of which can be directly attributed to the amount of energy imparted to the tissues. The currently used classification system aids in defining the degree of severity of these injuries and their subsequent risk for infection. The basic management principal for all of these injury patterns remains essentially the same, however: prevention of infection through debridement, wound management, antibiotic usage, and fracture stabilization. Frequently multiple surgical procedures will be required in order to obtain an infection free, united fracture with adequate soft tissue coverage (1). PMID- 16351593 TI - Antihypertensive treatment in patients with end-stage renal disease. AB - Arterial hypertension is a common finding in patients with end stage renal disease (80% patients are hypertensive). Cardiovascular diseases are the main cause of death in haemodialysis. The present study was performed to asses' successful treatment in hypertensive chronic haemodialysis patients by ultra filtration only and ultra filtration combined with medics. We studied 80 hypertensive adult patients who had been on regular haemodialysis treatment for at least 12 months (average duration of 41 months). All subjects were divided in two different antihypertensive treatment groups including 40 subjects each. The first group of patients were treated with trandolapril and ultra filtration, and the second group of patients were only treated with ultra filtration (control group). Blood pressure measurements before and after HD sessions were performed for each patient. Blood pressure control was defined using World Health Organization criteria 140/90 mm Hg. Average systolic blood pressure levels, after haemodialysis, were in the first group of patients 146.33 +/- 9.7 mm Hg, and in the control group 157,86 +/- 10.33 mm Hg. Average diastolic blood pressure was 87.83 +/- 8.11 mm Hg in the first group of patients and, in the control group it was 91.03 +/- 10.67 mm Hg. There were significant differences between systolic blood pressure level in the first group of patients and the control group of patients as well as in diastolic blood pressure (p < 0.05). We conclude that an antihypertensive therapy by trandolapril is more effective than ultra filtration alone in hypertensive patients on chronic haemodialysis. PMID- 16351594 TI - Clinical presentation of primary congenital hypothyroidism: experience before mass screening. AB - Primary congenital hypothyroidism is a common preventable cause of mental retardation. Neonatal thyroid screening is highly successful in early diagnosis and the improvement of developmental prognosis in the hypothyroid neonate. However, rarely cases could be missed, so doctors must be aware of the earl symptoms and signs of hypothyroidism. Therefore, the purpose of this study was to emphasize the presenting clinical features of primary congenital hypothyroidism at the age of diagnosis. The study population included 17 children with primary congenital hypothyroidism who attended the Department of Pediatrics, University Clinical Center Tuzla between 1986 and 1999. The diagnosis of all patients was confirmed by serum thyroid function tests (T4 and TSH). Of the 17 patients 10 (58.8%) were diagnosed in the first three months of life and 3 of them (17.6%) between fourth and sixth month of life. Four children (23.5%) were diagnosed after the age of six months. In the first three months of life hypothermia, constipation, jaundice, poor feeding, hoarse cry, macroglossia and hypoactivity were the moste common symptoms. Among the 17 patients with primary congenital hypothyroidism 5 of them (29.4 %) were diagnosed to have disgenetic thyroid tissue and 12 (70.6%) as having dyshormonogenesis. TSH and T4 levels were higher in patients in whom thyroid tissue was dysgenetic as comapared with those with dyshormonogenesis but the difference was not statistically significant (p > 0.05). Now it is expected that neonatal screening program in Bosnia and Herzegovina Federation will contribute to the detection of primary congenital hypothyroidism in early days of life. However, until an effective screening test is not yet routine in whole country, paediatricians should consider the diagnosis of hypothyroidism whenever it is clinically suggested. PMID- 16351595 TI - Obesity biological and psychological aspect. AB - There is no unique pattern to deal with obesity unless is presented as complex of biological and psychological factors. A lot of studies deal with only one side of it. This work shows both sides and discusses about all relevant factors, which are involved in pathogenesis of obesity. This is only way for finding better approaches for treatment and understandings for this issue. PMID- 16351596 TI - Bacterial aggregation in infected root canal. AB - The aim of this study was to investigate different microbial morphotypes in the root canal infection associated with chronic diffuse periapical lesion. In forty cases of asymptomatic teeth with radiographically diagnosed diffuse periapical lesion we took specimens of infected tissue from the root canals at the beginning of endodontic treatment. Fixation and four different staining methods of the specimens were obtained to provide microscope examination. All examined root canal specimens were heavily infected by bacteria. The most commonly identified were cocci 92 %, small mostly G+ diplococci and large G+cocci in clusters and grapelike groups, bacilli found in 67%, coccobacilli 37%, fungi 17%, and spirochetes in 5%. PMID- 16351597 TI - Evaluation of efficiency practical issues in the management of childhood asthma. AB - Asthma is the most common chronic illness in children and adolescents. The pediatric "Asthma School" has been established to improve the health and quality of life of children with asthma through education and support. The purpose of work is evaluation of efficiency education adolescents, children and parents in prevention, adequate use medications and control of asthma. It was operational investigation. The study was enrolled 70 participants from 7 cities in FBiH, which were attended "Asthma School". Evaluations of efficiency of education program have been done through questionnaires using 5-point scale. Observing variables were: attitude about quality of education and level of knowledge about asthma at the end of the educational round. "Asthma School" attended 62 parents and 8 adolescents. The age of children was 6-14 years. The most of participants were with middle school education (64.3%). Successfulness of seminar was score with highest point of scale by 79%. Access to health information was important for patients in adequate treating illness (80.6%) and in prevention of asthma (15%). There was a significant improvement for 38,4% in the basic knowledge of asthma among participants, from 2.7 before to 3.8 after education. Among 20 children that have learned proper technique breathing there was not register worsening symptoms of asthma by 83.4%. There was a significant improvement in the condition of patient, following by greater value PEF (72.3% participants). The results implicate necessity of continuity such action in order to make life of young asthma patient. PMID- 16351599 TI - Topical treatment of standardised burns with herbal remedies in model rats. AB - Experimental studies of burns require the use of different animal models. The aim of this work was to establish experimental model of thermal injuries and to evaluate the effects of topical agents on healing of the burn wounds. Forty female Wistar rats were randomly classified in 4 groups and isolated for 2 weeks before the onset of experiment. Animals were primarily anaesthetized with pentobarbital-sodium and then shaved (skin area of their back with diameters 5 cm x 5 cm). A round metal stamp with contact area of 5 cm2 and total weight of 100 g was heated up to 80 degrees C and then applied without additional pressure on the depilated skin of the back for 14 seconds. This procedure produced a standardized burn wound. Induced burn wounds were immediately drowned in the 4 degrees C- water for 3 s in order to maintain microcirculation. After the inducement of thermal injures, all rats were treated with 1% silver sulfadiazine cream, herbal topical preparations or were not treated at all. Burn wounds were treated twice a day until the healing completion. The result of treatment application was a significant reduction of burn wound diameters. Herbal topical preparations expressed positive therapeutic effects on the parameters of burn wounds. The efficiency of silver sulfadiazine cream in burn wound healing was significantly more expressed in comparison to healing process in control group of animals (p < or = 0,001). We conclude that herbal topical preparations efficiently caused the completion of burn wound healing process without scar formation. PMID- 16351598 TI - Foetal echocardiography. AB - A modern diagnostic and therapeutical approach to paediatric cardiology enables early application of foetal echocardiography in order of achieving diagnosis of congenital heart anomalies in utero. The aim of this study is to evaluate the percentage of prenatal diagnosis of congenital heart anomalies. This study has been conducted on 73 patients at Paediatric clinic of Clinical Centre of Sarajevo in a period from January 2000 until December 2004 with diagnosis of heart malformations. Among them 14 were preterm newborns, 40 boys. Diagnosis of cardiac anomalies with left to right shunt was done in 56.1%, obstructive 13.7%, cyanotic 1.36% and complex in 28.7% patients. The prenatal diagnosis was established in 4 patients (5.5%) by ultrasound examination which is very low in comparison to other European countries. There is a need for making prenatal diagnosis of congenital heart anomalies in foetus as early as it can be done in order to treat cardiac anomalies in utero, to decrease the number of congenital heart anomalies and to reduce the cost of cardiosurgical and postsurgical treatment. PMID- 16351600 TI - Posttraumatic seizures - prevention or not. AB - Early posttraumatic epilepsies (EPTE) are epileptic attacks that appear in first seven days after brain injury, with incidence of 3-5%. Predictors for development of EPTE are: impressive skull fracture with rupture of dura, intracranial haemorrhage, neurogical deficit (brain contusion), and posttraumatic amnesia longer than 24 hours. It is more common in children than in adolescents and adults. It carries four times increased risk for development of late posttraumatic epilepsy. Aspects of pharmacological prophylaxis was often considered, but scientifically neglected, without clear standings regarding controversial data in literature. Patients with severe head injury, hospitalised at Neurosurgical Hospital and Pediatric Hospital, Clinical Centre University of Sarajevo, in period from 6th of April 1992 till 1st of July 1994, were included in study. Prophylaxis of EPTE was carried out with phenobarbital (2-3 mg/kg) or phenytoin (3 mg/kg) parenterally. Decision was made upon clinical findings. CT scan was done in 13.5% patients, and in 31.9% patients serum concentrations of antiepileptic drugs were monitored. 310 patients aged 0-18 years (105 patients 0 10 years, and 205 patients 11-18 years) were investigated. Predictors of EPTE presented were posttraumatic amnesia longer than 24 hours in 90.6%, neurogical deficit in 86.45%, impressive skull fracture with rupture of dura in 81.3% and intracranial haemorrhage in 40.6%. Only two boys developed EPTE in first 24 hours after injury. This study has showed that use of antiepileptic drugs can decrease incidence of EPTE. However, problem remains, management of injured patients is still highly individualised, based on different experiences of doctors that treat patient, and without clear guidelines. PMID- 16351601 TI - Cytogenetic findings at Down syndrome and their correlation with clinical findings. AB - Down syndrome is a genetic state characterized by trisomy of chromosome 21. In the retrospective study for 12 years period (1991-2002) we have conducted correlation between cytogenetics analyses and clinical findings in our centre at 96 male and 83 female patients. Down syndrome was confirmed by cytogenetics analyses in 84 (87.5%) male patients and excluded in 12 (12.5%) male patients. Down syndrome was confirmed by cytogenetics analyses in 71 (85.5%) female patients and excluded in 12 (14.5%) female patients. Most common karyotype is free trisomy found in 139 (89.7%) examinees, than follows translocation form determined in 9 (5.8%), and mosaicism determined in 7 (4.5%) examinees. Our results indicate that cytogenetics analyses are necessary to confirm diagnosis of Down syndrome. PMID- 16351602 TI - Cystatin C as a marker for detection of early renal failure in diabetes type 2 patients. AB - The epidemiological studies have show dramatic increase and prevalence of end stage renal disease in patients with type 2 diabetes therefore early markers of diabetic nephropathy need to be identified (1). During the treatment of patients at the Clinic of endocrinology, diabetes mellitus and metabolic diseases in Sarajevo, we observed a necessity of application of new markers in assessment of early renal failure. Serum cystatin C level is another marker of renal function. Cystatin C is freely filtered at the level of the glomerulus and virtually all is re-absorbed and metabolized by the proximal tubular cells. Serum cystatin C is a screening test and an early indicator and predictor of the development of renal failure. AIM OF THE STUDY: To estimate correlation among cystatin C, serum creatinine and albuminuria in diabetes type 2 patients for assessment of early renal failure. Serum cystatin C as a screening test has to be considered in the treatment of diabetes type 2 patients for assessment of early renal failure. PMID- 16351603 TI - Models of treatment with antipsychotics of the schizophrenic patients. AB - The aim of this study were to determine which antipsychotic are currently in use, to establish which doses are administrated to patients, to find out is there a practice of proscribing simultaneously more then one antipsychotic drug, to determine whether antipsychotic are proscribed in divided doses, to establish whether there is, besides antipsychotics, treatment with other medicaments (co administration), especially with antiparkinsonics. The research (study) is epidemiological-clinical prospective, descriptive and analytical and it was conducted at University hospitals in Sarajevo, Tuzla and Mostar. Criteria for inclusion, non-inclusion and exclusion from the study were precisely defined as a mean for formation of sample. Based on this hypothesis were established, zero and alterative. According to zero hypothesis in the treatment of schizophrenia at University hospitals in FBiH new antipsychotic drugs are in use, small doses are proscribed (up to 20 mg), not more then one antipsychotic drug is used simultaneously, antipsychotics are administrated once a day and alongside with antipsychotics other medicaments are not co-administrated, especially antiparkinsons. The results of our study are showing that majority of patients are treated with classical antipsychotics. Minority of patients is treated with atypical neuroleptics like olanzapine, which is proscribed only in Sarajevo. Use of risperidone and ziprasidone is registered also only in Sarajevo, but only small number of patients is treated with these drugs. Most frequent antipsychotics were promazine and haloperidol. The range between minimal and maximal daily dose of promazine was from 50 to 450 mg/daily, and for haloperidol from 1 to 75 mg/daily. Above-mentioned drugs were administrated in an average from two to three times a day. Alongside with antipsychotics, other drugs were used. Most frequent was the use of biperidine in oral and parenteral formulation, as well as nitrazepam and diazepam. The importance of this study is following: data are useful for the current mental health care reform in FBiH, results will point out place and position of FBiH in contemporary world trends in the treatment of schizophrenia, they will contribute to rational use of antipsychotic therapy, they will point out possible ways in reduction of side effects, often dangerous adverse effects of antipsychotics, and they will give contribution to faster rehabilitation of schizophrenics with the reduction of financial means for the treatment of patients with schizophrenia. PMID- 16351604 TI - Structural differences between hip endoprostheses, and implications on a hip kinetics. AB - We compared kinetic characteristic of unipolar, bipolar and total hip endoprostheses, implanted after dislocated femoral neck fracture. Ninety patients were divided into three groups (30 patients in each group); a group with unipolar partial hip endoprosthesis (UPEP), a group with bipolar partial hip endoprosthesis (BPEP) and a group with total hip endoprosthesis (TEP). The patients from different groups were paired by parameters which could influence the long term functional result: follow up period, comorbidities, functional capabilities before injury, etc. After the average follow up 3.8 +/- 1.9 years, a measuring of range of hip motions (ROM) was conducted. The largest mean amplitudes in flexion (104 degrees), extension (13 degrees), abduction (35 degrees) and external rotation (38 degrees) was achieved BPEP, the largest adduction (14 degrees) was achieved UPEP, and internal rotation (34 degrees) TEP. Differences in ROMs are partially related to the clinical parameters such as: level of the hip pain, gait pattern, age and rehabilitation period (P < 0.05). Measuring of ROMs is the most reliable part of the clinical exam and it does not depend on subjectivity of patient, as opposed to other clinical parameters (level of pain, walking distance, aids usage, etc). The results obtained are favorable for the bipolar hip endoprosthesis, and they can be related to the biomechanical differences between the three types of hip endoprostheses. Kinetic advantages of the BPEP as compared to the UPEP, can be explained by the BPEP's structure: two level mobility and a thinner neck which delays impingement in the late motion phase. In comparison to the TEP, clinical advantages of the BPEP can be attributed to less extensive surgery and scarring. PMID- 16351605 TI - Listeria monocytogenes as the possible cause of the spontanous abortion in female of the fertile age. AB - Listeria monocytogenes is a Gram-positive, weakly pathogenic bacterium able to grow also at the temperature of 40 degrees C. A man most often gets affected by consuming contaminated food and water. Animals can carry bacteria although they have not to appear ill, then their meat and milk products are the source of infection of human being. The disease most often attacks with the weak immune system, newborns and pregnant women. That what is significant, listeria penetrates through the placenta and can lead to the fatal infection, which is characteristic by disseminated granulomatosis lesions of a newborn and micro abscess on the placenta. In a newborn can provoke the sepsis with the mortality of 50%. By the examination are encircled two groups of the reproductive age totally 60. From these 30 had one or more spontaneous abortion, and 30 had no spontaneous abortion. By the serologic reaction the agglutination is discovered the presence of the antibodies in serum of the. The positive serologic answer was found in the first group in 18 (60%) and in that another group in 8 participants (26, 70%). PMID- 16351606 TI - Erythema nodosum and lung ailments at the clinic for dermatological and venereal diseases in Sarajevo from 1989 to 2005. AB - In 15 years period (1989 - 2004) we have treated at our Clinic a total of 101 patients diagnosed with erythema nodosum which makes 2.9% of the total number of patients treated in this period. In order to explore correlation between dermatological and lung diseases we applied statistical analysis with respect to number, sex, age and irregularities occurring on the skin surface. Female patients dominated with 96 of them (95.6%) in the group, while only five patients were male (4.3%). This makes 22 : 1 scale in favor of female patients, as opposed to usual 5:1 ratio. Similar predominance (86%) was found in the study conducted by Mert and Gurkan (1.2). Average age of the patients was 41.6 years, which is slightly higher than the average of 18 and 34. The youngest patient was 17 and oldest 77. PMID- 16351607 TI - Prioritization of radiotherapy in Australia and New Zealand: where are we now? PMID- 16351608 TI - Review of non-positron emission tomography functional imaging of primary musculoskeletal tumours: beyond the humble bone scan. AB - Bone and soft tissue tumours are rare neoplasms. There are five major roles of imaging in the management of primary musculoskeletal tumours, that is, to differentiate between benignity and malignancy, to evaluate for local tumour extension, to screen for metastases, to judge the effect of chemotherapy, and to monitor for recurrence. To accomplish this, multiple modalities are required because no single examination is able to complete all these tasks. These modalities include plain radiography, CT, MRI, conventional nuclear medicine as well as positron emission tomography (PET) imaging. Elsewhere, PET imaging has been discussed at length, because it is likely to be superior in the assessment of bone and soft tissue tumours over conventional nuclear medicine procedures. However, conventional nuclear medicine may be of value when PET is unavailable. In this review, an overview of anatomical imaging will be given and the role of non-PET functional imaging will be discussed in detail. A variety of illustrative cases will be presented. PMID- 16351609 TI - Computed and conventional chest radiography: a comparison of image quality and radiation dose. AB - The aim of this study was to compare the image quality and entrance skin dose (ESD) for film-screen and computed chest radiography. Analysis of the image quality and dose on chest radiography was carried out on a conventional X-ray unit using film-screen, storage phosphor plates and selenium drum direct chest radiography. For each receptor, ESD was measured in 60 patients using thermoluminescent dosemeters. Images were printed on 35 x 43 cm films. Image quality was assessed subjectively by evaluation of anatomic features and estimation of the image quality, following the guidelines established by the protocols of the Commission of the European Communities. There was no statistically significant difference noted between the computed and conventional images (Wilcoxon rank sum test, P > 0.05). Imaging of the mediastinum and peripheral lung structures were better visualized with the storage phosphor and selenium drum technique than with the film-screen combination. The patients' mean ESD for chest radiography using the storage phosphor, film-screen combination and selenium drum was 0.20, 0.20 and 0.25 mGy, respectively, with no statistically significant difference with P > 0.05 (chi(2) tests). PMID- 16351610 TI - Endovascular treatment of arterial mesenteric ischaemia: a retrospective review. AB - Mesenteric ischaemia is a condition that has traditionally been managed surgically. It poses a challenging diagnostic and therapeutic problem, particularly in the acute setting. We review a small series of eight patients managed with endovascular techniques for either acute or chronic mesenteric ischaemia at The Royal Melbourne Hospital, from 1997 to 2002. We describe our results and relate these to the recent published literature regarding endovascular and surgical management of mesenteric ischaemia. Our experience confirms the valuable contribution of angioplasty and stenting in chronic mesenteric ischaemia, which compares favourably with surgery with regards to complication rates and mortality. We suggest the need for further studies to compare the long-term efficacy of endovascular techniques compared with surgery in the management of chronic mesenteric ischaemia. Furthermore, we demonstrate a role for endovascular management in acute mesenteric ischaemia, in the appropriate clinical setting. PMID- 16351611 TI - Ultrasound characteristics of histologically proven grade 3 invasive ductal breast carcinoma. AB - High-grade invasive ductal carcinoma (IDC) may paradoxically display features similar to benign breast masses. This study analysed the ultrasound features of histologically proven cases of grade 3 IDC. At North Western BreastScreen Victoria, from 4 February 1993 to 30 September 2002, 181 cases of grade 3 IDC that had ultrasound evaluation were available for retrospective analysis. For each tumour, four features were assessed: margin, attenuation characteristics, echotexture and depth versus width ratio. Eighty-seven per cent of tumours had an aggressive margin with an echogenic rind, microlobulation or angular margins. However, 11% had a well-defined smooth margin. The classic sonographic malignant feature of posterior shadowing was present in only 30%, whereas the tissues posterior to the lesion remained isoechoic in 48% and showed posterior enhancement in 11%. Six per cent of the tumours were isoechoic and difficult to appreciate on ultrasound. The best feature to characterize lesions as malignant was the margin of the lesion. To ensure that malignant lesions are correctly categorized, it is important that interfaces between the tumour and adjacent breast parenchyma are meticulously evaluated in a real-time fashion rather than viewed as a single still image. PMID- 16351612 TI - Managing waiting time for radiotherapy: a single machine unit experience. AB - Waiting time (WT) for radiotherapy (RT) is a significant clinical problem. This paper examines various strategies for managing WT for patients treated with radical and palliative intent in the new setting of a rural single machine unit in Australia. Cohorts of patients undergoing both radical and palliative RT in Bendigo had their WT prospectively recorded. Matched cohorts from the hub centre (Peter MacCallum Cancer Centre, Melbourne) treated with palliative intent were also collated. Strategies implemented included a devoted priority meeting, palliative points system, and reallocation of appointment times. The audit was to continue until best practice guidelines were bettered. Three cohorts of patients were compared. There is a significant trend for increasing numbers of patients treated per month since the centre opened (P < 0.0001). The ratio of palliative to radical intent patients remained stable between 46 and 52%. Mean WT for palliative RT reduced from 25 days in the first cohort to 7 days in the final cohort (P < 0.0005). Waiting time for palliative RT was initially longer at Bendigo than the hub centre (P < 0.0005), but by the final cohort there was a non significant difference favouring the Bendigo cohort (P = 0.26). Waiting time for radical treatment also improved throughout the three cohorts in Bendigo (P < 0.0005). A number of new strategies have successfully resulted in the abolition of lengthy WT for RT in Bendigo despite the increasing demand for the RT service. PMID- 16351613 TI - Prioritization of radiotherapy in Australia and New Zealand. AB - The purpose of this study was to document how radiation oncology departments in Australia and New Zealand manage extended waiting lists by prioritizing patients for radiotherapy and how these centres define the "waiting time". A literature search on strategies for management of waiting lists in radiotherapy, both locally and internationally, was performed. A collaborative survey of all the radiotherapy departments in Australia and New Zealand was then undertaken. Of the 32 centres surveyed around Australia and New Zealand, 25 (77%) responded. There was considerable variation in the definitions used for "waiting times". Eleven of the 25 centres had formally documented protocols. New Zealand has a national policy for prioritization of patients for radiotherapy. Six centres had verbal protocols. Four centres had no significant waiting times and did not require a protocol for prioritization. One centre prioritized according to clinician discretion, two centres used a first-come, first-served basis. One centre replied but their protocol was missing. The variation in the definition of waiting time reduces its usefulness as an indirect measure of resources and as a method of comparing centres. There is also wide variation in the management of waiting lists, particularly in the prioritization schedules used by different centres. The major factor contributing to waiting lists at present is a shortage of radiation oncology staff, particularly radiation therapists. The implementation of standardized protocols for prioritizing patients may be useful in helping to manage scarce resources not withstanding the need to increase the resource base. However, the existence of such protocols should not give legitimacy to undue delays in commencing radiation treatment. PMID- 16351614 TI - Bilateral optic pathway glioma with intracranial calcification: magnetic resonance imaging and magnetic resonance spectroscopy findings. AB - Optic nerve glioma is the most common primary neoplasm of the optic nerve in childhood. It can extend intracranially along the optic pathway (optic pathway glioma). The lesion tends to present with decreased visual acuity in the affected eye, but can cause additional symptoms when it is large. Local involvement within the orbit can be characterized using CT, but MRI is superior in showing the intracranial extent of the lesion. Intracranial calcification in optic pathway glioma is rare. We present a rare case of optic pathway glioma with calcification in the intracranial component. Also, we describe MR spectroscopy (MRS) findings in this case. PMID- 16351615 TI - Multiple pancreatic metastases from clear cell renal carcinoma: diagnosis with chemical shift magnetic resonance imaging before surgery. AB - We present a case in which multiple pancreatic tumours were diagnosed as metastatic clear cell renal carcinomas with chemical shift MRI (CSI) before surgery. Radiologists may be unable to recognize the loss of intensity on CSI macroscopically. We believe that it is useful to make subtraction images and calculate signal intensity on CSI, even if the lesions are multiple metastatic tumours. PMID- 16351616 TI - Intracranial metastasis from prostatic adenocarcinoma simulating a meningioma. AB - We report an unusual case of extra-axial metastatic adenocarcinoma of the prostate that closely simulated a frontal, parasagittal, dural-based meningioma. Such tumours, which satisfy several criteria for a diagnosis of meningioma, but which have proved instead to be metastatic adenocarcinoma of the prostate, form the focus of our report. PMID- 16351617 TI - Paradoxical embolization via a patent foramen ovale following acute pulmonary embolism. AB - The foramen ovale is usually obliterated following establishment of the adult circulation but remains patent in 25% of individuals. This potential communication between the venous and arterial circulations can allow thromboembolic material to bypass the lungs and enter the systemic circulation. We report two cases of paradoxical embolization through a patent foramen ovale following acute large pulmonary embolism (PE) and discuss the factors that predispose to paradoxical embolization following PE. PMID- 16351618 TI - Paget's disease of the bone in a Chinese woman. AB - Paget's disease, otherwise known as osteitis deformans, is an unusual condition in the Oriental population. We report a case of Paget's disease in a Chinese woman, incidentally diagnosed on a bone scan. This was confirmed by clinical history, biochemistry and imaging findings using other modalities. Although bone scans are commonly performed to diagnose traumatic occult fractures and bone metastases, they can also be used to diagnose metabolic bone diseases, such as osteoporotic fractures, and to evaluate Paget's disease. It can also diagnose acute fractures secondary to renal osteodystrophy and osteomalacia. PMID- 16351619 TI - Movement of the side hole occurring in a port-catheter system percutaneously implanted for hepatic arterial infusion chemotherapy. AB - The authors report a case in which catheter-related difficulties occurred with a port-catheter system implanted to perform repeated hepatic arterial infusion chemotherapy for advanced hepatocellular carcinomas. The port-catheter system had been percutaneously implanted with the catheter tip fixation method, and a side hole had been created in the indwelling catheter for placement in the common hepatic artery. Four months after port-catheter placement, however, the indwelling catheter became bent at the location of the side hole in the proper hepatic artery. Subsequently, the side hole moved into the left hepatic artery, resulting in a lack of distribution of chemotherapeutic agents to the tumours in the segments of the liver supplied by the right hepatic artery. This situation was easily corrected with a simple interventional radiological method so that hepatic arterial infusion chemotherapy could be resumed with sufficient distribution over the entire liver. To our knowledge, such a problem involving a port-catheter system implanted with the catheter tip fixation method and its management thereafter has not been reported in the published literature. PMID- 16351620 TI - Pre and postnatal diagnosis of fetus-in-fetu. AB - Differentiation of abdominal masses detected on prenatal ultrasound is difficult and requires careful characterization of the mass and precise localization. Differentiation is required in order to distinguish benign from potentially malignant conditions. We describe a case of fetus-in-fetu with pre- and postnatal imaging. PMID- 16351621 TI - Glomus tumour in a child. AB - Glomus tumours are uncommon neoplasms. Very rarely do they present in the paediatric age group. We describe a case of bilateral glomus tumours in a 13-year old girl that were successfully treated with radiotherapy. The patient remains well 8 years after completion of treatment. PMID- 16351622 TI - Carcinoma of the tonsil with bilateral breast metastasis: a rare presentation. AB - With increasing local regional control and survival rates, the incidence of distant metastasis is increasing. Head and neck malignancies traditionally metastasize to the lung, liver and bone. We present a case of an elderly woman who was loco-regionally controlled after radiotherapy for carcinoma of the tonsil, but later presented with breast metastasis. To our knowledge, this metastatic pattern has never been reported in published literature. We also discuss the management of breast metastasis and the need to avoid unnecessary mastectomies in such patients. PMID- 16351623 TI - RE: Another technique for radiation treatment of the supraorbital nerve. PMID- 16351624 TI - RE: Chart round in a skin radiotherapy unit. PMID- 16351630 TI - Renal low-signal intensity on magnetic resonance imaging in a case of paroxysmal nocturnal haemoglobinuria. PMID- 16351631 TI - All-trans retinoic acid-induced myositis. PMID- 16351632 TI - Protecting against anthracycline-induced myocardial damage: a review of the most promising strategies. AB - Over the last 40 years, great progress has been made in treating childhood and adult cancers. However, this progress has come at an unforeseen cost, in the form of emerging long-term effects of anthracycline treatment. A major complication of anthracycline therapy is its adverse cardiovascular effects. If these cardiac complications could be reduced or prevented, higher doses of anthracyclines could potentially be used, thereby further increasing cancer cure rates. Moreover, as the incidence of cardiac toxicity resulting in congestive heart failure or even heart transplantation dropped, the quality and extent of life for cancer survivors would improve. We review the proposed mechanisms of action of anthracyclines and the consequences associated with anthracycline treatment in children and adults. We summarise the most promising current strategies to limit or prevent anthracycline-induced cardiotoxicity, as well as possible strategies to prevent existing cardiomyopathy from worsening. PMID- 16351633 TI - Childhood acute lymphoblastic leukaemia and relapse. AB - Acute lymphoblastic leukaemia (ALL) is the most common childhood cancer. Treatment has improved but relapsed ALL remains more common than new cases of many 'common' paediatric malignancies. We have salvage regimens with substantial complete remission (CR) rates and increasing access to haematopoietic stem cell transplantation, but most patients who relapse die. We need better therapies. Insights into pharmacology may guide more effective use of existing agents. Novel agents with activity against resistant lymphoblasts offer an appealing strategy. However, most candidate agents fail, despite enthusiastic investigators, intriguing mechanisms of action and 'compelling' preclinical data. A number of existing combinations provide a 40% complete response rate in second or third relapse. Yet survival in third remission is <10%. Novel agents must, most likely, be integrated into multiagent combinations that provide a higher CR rate or better quality CR's than our conventional combinations in order to contribute substantially to cure. The march from bench to bedside requires careful consideration of the intermediate steps. PMID- 16351634 TI - Platelet transfusion prophylaxis for patients with haematological malignancies: where to now? AB - National guidelines for platelet transfusion in many countries recommend that the general platelet transfusion trigger for prophylaxis is 10x10(9)/l. This annotation reviews the evidence for this threshold level and discusses other current unresolved issues relevant to platelet transfusion practice such as the optimal dose and the clinical benefit of a strategy for the prophylactic use of platelet transfusions when the platelet count falls below a given threshold. PMID- 16351635 TI - High expression of calcium-binding proteins, S100A10, S100A11 and CALM2 in anaplastic large cell lymphoma. AB - Anaplastic large cell lymphomas (ALCL) are characterised by the presence of CD30 positive large cells, which usually are of T-cell type. Based on the presence or absence of translocations involving the anaplastic lymphoma kinase (ALK) locus, ALCL cases can be divided into two groups. To gain more insight in the biology of ALCL, we applied serial analysis of gene expression (SAGE) on the Karpas299 cell line and identified 25 up- and 19 downregulated genes. Comparison of the differentially expressed genes with DNA copy number changes in Karpas299 revealed that two overexpressed genes, S100A10 and S100A11, were located in an amplicon suggesting that the increased mRNA levels were caused by DNA amplification. Quantitative reverse transcription polymerase chain reaction on 5 ALCL cell lines and 12 ALCL tissues confirmed the SAGE data for 13 out of 14 up- and one out of four downregulated genes. Immunohistochemical staining confirmed the presence of S100A10, a calcium-binding protein, in three out of five ALK+ and all 7 ALK- ALCL cases. S100A11 staining was confirmed in all ALK+ and six of seven ALK- ALCL cases. Three of the upregulated genes represented calcium-binding proteins, which suggest that altered intracellular signaling might be associated with the oncogenesis of ALCL. PMID- 16351636 TI - A non-randomised dose-escalating phase II study of thalidomide for the treatment of patients with low-risk myelodysplastic syndromes: the Thal-SMD-2000 trial of the Groupe Francais des Myelodysplasies. AB - Patients (n=47) with low-risk myelodysplastic syndrome were treated with thalidomide [200 mg/d, increased by 200 mg/d/4 weeks up to week 16]. Responses were evaluated according to the International Working Group criteria at week 16 for 39 patients who received at least 8 weeks of treatment. Twenty-three (59%) patients showed haematological improvement (HI): four major erythroid response (HI-EM), 15 minor erythroid response, six major neutrophil response, two major platelet response. Side effects caused 22/39 to stop thalidomide before week 16. Nine of 23 responders continued thalidomide after week 16 [19% of trial patients] with sustained response in eight of nine. Six reached week 56, including the four HI-EM patients [13% of trial patients]. Nineteen of 36 red blood cell transfusion dependent patients (53%) showed erythroid response, but only four became transfusion-independent. Among the 23 responders, the median duration of response was 260 d (range 30-650). Responses were sustained in all patients except one, and were observed between week 4 and week 8 in 85% of patients, at doses ranging from 200 to 400 mg. Only two patients responded at 600 mg/d and none at 800 mg/d. No clinical characteristics of responding versus non-responding patients were identified. The erythroid response rate was identical in all cytogenetic subgroups, including 5q31.1 deletions. Pretreatment vascular endothelial growth factor levels were lower in responders compared with non-responders (P=0.004). Microvessel density (MVD) increased and apoptosis decreased in four of six and in all six responders studied respectively whereas MVD and apoptosis were unchanged in three non-responders. PMID- 16351637 TI - Monitoring gastric lymphoma in peripheral blood by quantitative IgH allele specific oligonucleotide real-time PCR and API2-MALT1 PCR. AB - Gastric extranodal marginal zone lymphoma (EMZL) often shows prolonged localised disease, but the present study demonstrated the presence of tumour cells in peripheral blood (PB) of low stage patients. We studied the presence of tumour cells in PB in gastric lymphoma patients harbouring or lacking t(11;18)(q21;q21), by real-time immunoglobulin (Ig)H allele-specific oligonucleotide-polymerase chain reaction (ASO-PCR) and API2-MALT1 PCR. Tumour cells were exclusively detected in PB of t(11;18)(q21;q21)+-EMZL patients. The presence of tumour cells in PB and gastric biopsy follow-up samples showed a good correlation in these patients, suggesting clinical relevance for monitoring of tumour cells in PB of gastric t(11;18)(q21;q21)+-EMZL patients. PMID- 16351638 TI - Childhood anaplastic large cell lymphoma has a high incidence of ALK gene rearrangement as determined by immunohistochemical staining and fluorescent in situ hybridisation: a genetic and pathological correlation. AB - Anaplastic large cell lymphoma (ALCL) comprises 10-15% of childhood non-Hodgkin lymphomas (NHL). Systemic ALCL is highly associated with anaplastic lymphoma kinase (ALK) gene translocations with over-expression of ALK protein. We studied ALK rearrangements using fluorescence in situ hybridisation (FISH) and ALK immunohistochemical staining in 43 paediatric systemic ALCLs. FISH (performed on 35 cases) identified a translocation in 29 cases (83%). Immunohistochemistry identified ALK over-expression in 42/43 cases (97%) with the single ALK-negative case demonstrating an ALK rearrangement by FISH, indicating 100% incidence of ALK translocations. PMID- 16351639 TI - Up-regulation of the T cell quiescence factor KLF2 in a leukaemic T-cell line after expression of the inositol 5'-phosphatase SHIP-1. AB - Inositol 5'-phosphatase SHIP-1 (SHIP) is a negative regulator of signal transduction in haematopoietic cells. SHIP inactivation may be involved in the pathogenesis of leukaemia. An inducible expression system was combined with microarray analysis to identify target genes regulated by SHIP in the human T cell leukaemia cell line Jurkat. One gene identified was Kruppel-like factor 2 (KLF2), which was up-regulated two to threefold at the RNA and protein level after the induced expression of SHIP. KLF2, a negative regulator of T cell proliferation, has been implicated in T cell quiescence. KLF2 or SHIP expression in Jurkat cells caused 45% or 60% reduction of proliferation, respectively. SHIP can up-regulate KLF2 expression, implicating KLF2 in the SHIP-mediated growth inhibition of a human leukaemic T-cell line. PMID- 16351640 TI - Serial studies of methylation of CDKN2B and CDKN2A in relapsed acute promyelocytic leukaemia treated with arsenic trioxide. AB - Ninety consecutive patients with acute promyelocytic leukaemia were investigated for promoter methylation of CDKN2B (alias p15) and CDKN2A (alias p16) in disease relapse and progression. CDKN2B methylation was significantly more frequent at first relapse (30/36, 83%) than at presentation (48/77, 62%) (P=0.025), while CDKN2A methylation appeared unaffected. Both acquisition and loss of CDKN2B methylation happened at relapse, with acquisition more frequent. No significant increase in CDKN2B and CDKN2A methylation occurred at more advanced relapses. At first or subsequent relapses, owing to highly effective salvage by arsenic trioxide, CDKN2B methylation did not impact on event-free survival or overall survival. PMID- 16351641 TI - Congenital amegakaryocytic thrombocytopenia: a retrospective clinical analysis of 20 patients. AB - Congenital amegakaryocytic thrombocytopenia (CAMT) is a rare bone marrow failure syndrome characterised by isolated thrombocytopenia because of ineffective megakaryocytopoiesis at birth. In the last 10 years, we collected data from 20 patients diagnosed with CAMT based on a severe thrombocytopenia since birth and absent or markedly decreased numbers of megakaryocytes in the bone marrow. Fanconi's anaemia and thrombocytopenia absent radii syndrome were ruled out for all patients. We retrospectively compared the clinical courses, laboratory findings and treatment outcome. Development of pancytopenia was observed in 14 of the patients, only one patient presented with an isolated thrombocytopenia over a period of over 14 years. One boy died from bleeding complications. We defined two groups of patients according to the course of platelet counts during the first year of life, which also differed in the course of development of pancytopenia. Physical anomalies in addition to haematopoiesis were found in a number of patients: two children presented with cardiac defects, six with growth abnormalities, and four with retardation of psychomotor development. Fifteen patients were treated with haematopoietic stem cell transplantation, four of whom died of transplantation-related events. PMID- 16351642 TI - High dose factor VIIa improves clot structure and stability in a model of haemophilia B. AB - Factor IX (FIX) deficiency results in haemophilia B and high dose recombinant activated factor VII (rFVIIa) can decrease bleeding. Previously, we showed that FIX deficiency results in a reduced rate and peak of thrombin generation. We have now used plasma and an in vitro coagulation model to examine the effect of these changes in thrombin generation on fibrin clot structure and stability. Low FIX delayed the clot formation onset and reduced the fibrin polymerisation rate. Clots formed without FIX were composed of thicker fibrin fibres than normal. rFVIIa shortened the clot formation onset time and improved the fibre structure of haemophilic clots. We also examined clot formation in the presence of a fibrinolytic challenge by including tissue plasminogen activator or plasmin in the reaction milieu. In these assays, normal FIX levels supported clot formation; however, clots did not form in the absence of FIX. rFVIIa partially restored haemophilic clot formation. These results were independent of the effects of the thrombin-activatable fibrinolysis inhibitor. Our data suggest that rFVIIa enhances haemostasis in haemophiliacs by increasing the thrombin generation rate to both promote formation of a structurally normal clot and improve clot formation and stability at sites with high endogenous fibrinolytic activities. PMID- 16351643 TI - Role of interleukin-6 in hypoxic regulation of intestinal iron absorption. AB - The regulation of intestinal iron absorption is not fully understood. Hepcidin, a liver-produced peptide, has recently been identified as a negative regulator of iron absorption in various conditions associated with altered iron metabolism (e.g. inflammation, anaemia, hypoxia). It is not clear whether these perturbants share a common signalling pathway. In this study, the importance of the cytokine interleukin-6 (IL-6) was investigated in the hypoxic mouse model. Hypoxia was associated with increased levels of circulating IL-6, decreased liver hepcidin mRNA and increased iron absorption (especially MT). A significant positive correlation existed between the total iron uptake and IL-6 levels in circulation. IL-6 per se, though inducing hepcidin mRNA, failed to affect basal iron absorption. The adaptive response to absorption following the hypoxic exposure was, however, more prominent if mice had been treated concurrently with IL-6. This enhancement in absorption occurred even though hepcidin mRNA was not significantly changed. Similar prominent responses were seen with both human and mouse IL-6. Anti-IL-6 antiserum normalised iron absorption in mice exposed to hypoxia, because of a reduction in the MT. These data indicate that IL-6 can influence iron absorption (especially MT) during the hypoxic exposure, but via a mechanism independent of hepcidin. PMID- 16351644 TI - Genetic and clinical heterogeneity of ferroportin disease. AB - Ferroportin is encoded by the SLC40A1 gene and mediates iron export from cells by interacting with hepcidin. SLC40A1 gene mutations are associated with an autosomal type of genetic iron overload described as haemochromatosis type 4, or HFE4 (Online Mendelian Inheritance in Man number 606069), or ferroportin disease. We report three families with this condition caused by novel SLC40A1 mutations. Denaturing high-performance liquid chromatography was employed to scan for the SLC40A1 gene. A D181V (A846T) mutation in exon 6 of the ferroportin gene was detected in the affected members of an Italian family and shown to have a de novo origin in a maternal germinal line. This mutation was associated with both parenchymal and reticuloendothelial iron overload in the liver, and with reduced urinary hepcidin excretion. A G80V (G543T) mutation in exon 3 was found in the affected members of an Italian family with autosomal hyperferritinaemia,. Finally, a G267D (G1104A) mutation was identified in exon 7 in a family of Chinese descent whose members presented with isolated hyperferritinaemia. Ferroportin disease represents a protean genetic condition in which the different SLC40A1 mutations appear to be responsible for phenotypic variability. This condition should be considered not only in families with autosomal iron overload or hyperferritinaemia, but also in cases of unexplained hyperferritinaemia. PMID- 16351648 TI - The prognostic value of p53, Ki-67 and matrix metalloproteinases MMP-2 and MMP-9 in transitional cell carcinoma of the renal pelvis and ureter. AB - AIM: To investigate the prognostic and predictive relevance of p53 protein, Ki-67 antigen, MMP-2 and MMP-9 in patients with transitional cell carcinoma (TCC) of the upper urinary tract. METHODS: The expression of p53 protein, Ki-67 antigen, MMP-2 and MMP-9 was examined by immunohistochemistry in 69 patients with TCC of the upper urinary tract. Correlation of p53, Ki-67, MMP-2 and MMP-9 over expression with conventional pathological parameters and patient survival was examined. RESULTS: p53 over-expression was significantly correlated with histological grade (P < 0.05), but not with pathological stage, vascular invasion, lymphatic invasion or lymph node metastasis. Ki-67 over-expression was significantly correlated with stage, grade, lymphatic invasion and vascular invasion (P < 0.05). In survival analyses, Ki-67 over-expression was a significant prognostic factor in the univariate analysis (P < 0.05), but it did not have a significant impact on survival in the multivariate analysis. Ki-67 labeling index was a significant prognostic factor in patients with a low p53 labeling index, but not in patients with a high p53 labeling index. CONCLUSION: Ki-67 over-expression is of prognostic value in TCC of the upper urinary tract, while p53, MMP-2 and MMP-9 are of limited value. PMID- 16351649 TI - Identification and validation of risk factors for vesicourethral leaks following radical retropubic prostatectomy. AB - AIMS: To identify risk factors for anastomotic leaks at cystography following radical retropubic prostatectomy (RRP). METHODS: In phase 1 the records of a 107 RRP patients were reviewed. Data collected included comorbidity, pathological factors and intra- and postoperative complications. From these, risk factors were identified that were associated with a leak at cystography. In phase 2 (n = 46) we prospectively tested if the risk factors identified could predict an anastomotic leak. RESULTS: In phase 1 the only identifiable risk factors were that of a difficult anastamosis, an unsatisfactory intraoperative test flush and the presence of a urinary tract infection. One or more of these events were found in 17/25 (68%) of the patients who leaked (P < 0.0001). Of the eight leaks missed, five were classed as minimal and did not require repeat cystography. Within the prospective phase 2 cohort one or more risk factors were present in 7/10 (70%) of the patients who leaked. In contrast, the identified risk factors were present in only 5/36 (13.8%) of the patients who did not leak (P < 0.001). The specificity of the test was 86.1% with a sensitivity of 70%. This gave a positive predictive value of 58.3% and a negative predictive value of 91.1%. Three leaks (two minimal and one moderate) would have been missed but 31 (67.3%) patients would have avoided an unnecessary radiological study. CONCLUSION: Using a retrospective and prospective cohort of patients we have shown that a cystogram following RRP can be safely avoided in the absence of the identified risk factors. PMID- 16351650 TI - Impact of radical perineal prostatectomy on urinary continence and quality of life: a longitudinal study of Japanese patients. AB - AIM: We used self-completed questionnaires to obtain a longitudinal assessment of urinary continence and urinary, bowel, and sexual domain-related quality of life (QOL) in Japanese patients undergoing radical perineal prostatectomy (RPP). METHODS: A total of 41 Japanese patients with a median age of 69 years who underwent RPP between February 2002 and February 2004 were included in the study. We measured QOL by the University of California, Los Angeles, Prostate Cancer Index (UCLA-PCI) and assessed urinary continence on the basis of three different definitions of continence. The International Prostate Symptom Score (I-PSS) was also included to evaluate lower urinary tract symptoms (LUTS). RESULTS: When urinary continence was de fi ned as none, one, or two protective pads per day, 100%, 73%, 94%, 97%, or 100% of the patients were continent before, and 1, 3, 6, and 12 months after, RPP, respectively. When it was de fi ned as total control or occasional dribbling, the corresponding values were 97%, 70%, 84%, 94%, and 97%. Urinary function returned to the preoperative baseline level by 6 months postoperatively and scores for urinary bother had significantly surpassed the baseline by 12 months (P = 0.043). The I-PSS was significantly improved (P = 0.014), with a mean 4.7-unit decrease. Sexual function worsened significantly after surgery, and its recovery was less favorable. No significant change was observed in scores for bowel function or bowel bother. CONCLUSIONS: The majority of patients who undergo RPP rapidly regain urinary continence and QOL within 3-6 months. RPP has a favorable impact on LUTS. PMID- 16351651 TI - Extensive biopsy using a combined transperineal and transrectal approach to improve prostate cancer detection. AB - PURPOSE: Previous studies have indicated that 6-core transrectal prostate biopsy misses a considerable number of cancers. We performed an extensive biopsy protocol of 12-core sampling using both transperineal and transrectal approaches to determine the impact on the cancer detection rate. MATERIALS AND METHODS: We prospectively evaluated 402 men who underwent 6-core transperineal and 6-core transrectal biopsies simultaneously due to abnormal digital rectal examination (DRE) and/or elevated prostate-specific antigen (PSA) levels of 4.0 ng/mL or greater. Using the transperineal approach we obtained four cores from the bilateral peripheral zone targeting the lateral and parasagittal areas and two cores from the bilateral transition zone. The following transrectal biopsy was performed traditionally. We compared cancer detection rate between the extended 12-core procedure and conventional 6-core transperineal and transrectal groups in terms of total PSA and DRE findings. RESULTS: Using the extensive combined method, prostate cancer was detected in 195 cases (48.5%) and the detection rate significantly increased 7.2% and 8.5% compared to the transperineal and transrectal groups, respectively. According to PSA levels and DRE findings, the cancer detection rate by the combined method was significantly improved in patients with PSA levels of 4-10 ng/mL and negative DRE: 10.3% and 11.6% compared to the transperineal and transrectal groups, respectively. CONCLUSIONS: The extensive 12-core method significantly improved the overall cancer detection rate and was especially efficient for men with PSA levels of 4-10 ng/mL accompanied by a negative DRE finding. PMID- 16351652 TI - Significance of routine transition zone biopsies in Japanese men undergoing transrectal ultrasound-guided prostate biopsies. AB - BACKGROUND: The objective of this study was to evaluate the clinical significance of additional routine transition zone (TZ) biopsies in Japanese men undergoing transrectal ultrasound (TRUS)-guided systematic 8-core peripheral zone (PZ) biopsies. METHODS: Between October 2002 and December 2004, a total of 788 consecutive patients underwent TRUS-guided systematic biopsy of the prostate for the fi rst time. As a rule, 10 cores were taken from each patient; that is, 8 cores from the PZ, including the standard sextant cores and 2 cores from the anterior lateral horns, and 2 additional cores from the bilateral TZ. The cancer detection rate was calculated according to several parameters. We also assessed the disease extent on radical prostatectomy specimens according to the cancer location within the biopsy specimens. RESULTS: Prostate cancer was detected by 10 core biopsies in 209 (26.5%) of the 788 patients, and 11 of these patients had positive cores only in the TZ; that is, the increase in cancer detection rate by sampling two additional cores from the TZ was 5.3%. Among 209 patients diagnosed as having prostate cancer, radical prostatectomy without any neoadjuvant therapy was performed in 59 patients with positive biopsy cores in the PZ, 7 in the TZ and 32 in both the PZ and TZ. Patients with positive cores in both zones showed significantly less favorable characteristics, indicating more advanced disease than that in those with positive cores in either zone. CONCLUSIONS: Routine TZ biopsy did not significantly increase the detection rate of prostate cancer; however, the anatomical location of positive biopsy cores could provide additional information concerning disease extension in patients undergoing radical prostatectomy. PMID- 16351653 TI - Prostate gland volume is a strong predictor of biopsy results in men 70 years or older with prostate-specific antigen levels of 2.0-10.0 ng/mL. AB - PURPOSE: The incidence of prostate cancer, benign prostatic enlargement and serum level of prostate-specific antigen (PSA) increase with patient age. Intermediate elevation of PSA in elderly populations is apt to be considered insignificant. We evaluated the impact of PSA and prostate volume on the presence of non-palpable prostate cancer in elderly men with an intermediate level of PSA. MATERIALS AND METHODS: Clinical records of 154 men 70 years or older, with non-cancerous digital rectal examination findings and with serum PSA levels of 2.0-10.0 ng/mL, who underwent initial 6- to 10-core transrectal prostate biopsy, were reviewed for prostate volume, number of biopsy cores, PSA and associated parameters. Stepwise logistic regression and receiver operating characteristic (ROC) models were used to determine the impacts of the parameters on the biopsy results. RESULTS: Overall cancer detection rate was 40/154 or 26.0%. Prostate-specific antigen showed no significant association with the presence of prostate cancer (P = 0.59, Mann-Whitney U-test), while prostate volume did (P < 0.0001). On stepwise logistic regression analysis, prostate volume (P = 0.024, 95% CI 1.008-1.116) and biopsy core density (P = 0.017, 95% CI 4.76-7.12 x 10(6)) were independently associated with a cancer diagnosis, whereas PSA density was not an independent factor for a positive biopsy result. The area under the ROC curve for prostate volume was significantly superior to that of PSA (0.802 vs. 0.529; P = 0.012). CONCLUSIONS: In men 70 years or older with gray zone PSA, prostate cancer patients are equally distributed over any PSA range. Although PSA has less impact on cancer presence than mere prostate volume, prostate cancer would be detected in a substantial proportion of older patients with PSA levels of 2.0-10.0 ng/mL. PMID- 16351654 TI - Factors influencing practices for chronic prostatitis: a nationwide survey of urologists in South Korea. AB - BACKGROUND: We investigated the influence of physicians' attitudes on the treatment of chronic prostatitis, and attempted to correlate demographic and professional characteristics with patterns of therapy regarding chronic prostatitis. METHODS: Probability samples were drawn from the Korean Urological Association Registry of Physicians, and a random sample of 850 Korean urologists were asked to complete a questionnaire which explored practice characteristics, attitudes, and diagnostic and treatment strategies for the management of chronic prostatitis. The returned questionnaires were statistically analysed. RESULTS: Of the 850 questionnaires sent, 302 were returned, and 275 of those were included in the final analysis (response rate 32.4%). Multivariate logistic regression analysis indicated that the type of hospital (P < 0.001) and belief that culture tests helped diagnose chronic prostatitis (P = 0.001) were the most determinant factors with respect to the routine performance or non-performance of culture tests. Most Korean urologists (96.4%) prescribed antibiotics for the primary treatment of chronic prostatitis. Even when primary antibiotic treatment was unsuccessful, urologists frequently prescribed a second course of antibiotics (57.8%). In the multivariate model used, the likelihood of prescribing antibiotics as a secondary treatment varied by the type of hospital, with 0.4 fold odds (odds ratio [OR], 0.43; 95% confidence interval [CI], 0.21-0.87; P = 0.019) of the practice occurring in university hospitals, as opposed to primary clinics. According to the same model, the belief that culture tests help to diagnose chronic prostatitis was also an independent influencing factor (OR, 2.29; 95% CI, 1.39-3.77; P = 0.001), whereas the actual performance of culture tests had no statistical significance. CONCLUSION: Our findings suggest that the personal beliefs and professional characteristics of physicians may influence the diagnosis and treatment of chronic prostatitis in Korea. PMID- 16351655 TI - Is there a possible correlation between chromosomal variants and spermatogenesis? AB - AIM: Heterochromatin polymorphism is considered a variant of a normal karyotype but is more frequent in infertile men. The aim of this study was to evaluate the correlation between heterochromatic variants and male infertility and to discuss the possible mechanisms of how heterochromatic polymorphism might affect spermatogenesis. METHODS: Cytogenetic analysis was undertaken in 210 infertile males who had been taken into assisted reproductive techniques and in 183 men with proven fertility. Additionally, C-banding was performed in men with heterochromatin polymorphism. Sperm fluorescence in situ hybridization (FISH) was applied in 54 men with normal karyotype presenting either normal or abnormal sperm parameters as well as in 8 men with heterochromatin polymorphism. The outcomes of assisted reproductive techniques were compared between infertile men with normal karyotype and men with heterochromatin polymorphism. RESULTS: The incidence of heterochromatin polymorphism was higher in infertile men. The most frequent chromosome involved in heterochromatin polymorphism was chromosome 9. Sperm FISH analysis revealed an increased rate of aneuploidy in men with heterochromatin polymorphism. Laboratory and clinical outcomes in assisted reproductive techniques were compromised in men with heterochromatin polymorphism. CONCLUSIONS: An increased rate of heterochromatin polymorphism in infertile males seems to be more than an incidental finding, and must not be considered as a normal variant. Polymorphic heterochromatin may have deleterious effects on the genetic constitution of spermatozoa. More attention must be directed to infertile men with heterochromatin polymorphism. PMID- 16351656 TI - The effect of partial unilateral ureteral obstruction release and allopurinol on the renal malondialdehyde and glutathione levels. AB - PURPOSE: The aim of the present study was to evaluate whether relief of partial unilateral ureteral obstruction (PUUO) with or without antioxidant drug affect renal tissue malonedialdehyde (MDA) and glutathion (GSH) levels. MATERIALS AND METHODS: A total of 25 rats were used in this PUUO study. Partial unilateral ureteral obstruction was created by the burial of the upper one-third of the left ureter in the psoas muscle. The rats were sacrificed on 28th day following PUUO. Relief of the obstruction was performed twenty minutes before sacrifice by cutting the proximal ureter in reperfusion group. 50 mg/kg intraperitoneal allopurinol was administered 20 minutes before relief of obstruction in the antioxidant group. Renal tissue MDA and GSH levels were measured in both kidneys. RESULTS: At the end of the study 5, 7 and 7 rats could only be interpreted in sham, reperfusion and antioxidant groups, respectively. While the mean left and right renal MDA and GSH levels were statistically different from each other in reperfusion group (P < 0.001), there were no significant differences in the sham (P > 0.05) and antioxidant (P > 0.05) group. Both the mean sham group left and right renal tissue MDA or GSH levels were significantly different from reperfusion group, but only the mean sham group left renal tissue MDA and right renal tissue GSH levels were not statistically different from antioxidant group (P < 0.05). The mean left or right renal MDA and GSH tissue levels of the antioxidant group were statistically different from reperfusion group (P < 0.05) except for the right renal tissue GSH level (P > 0.05). CONCLUSION: Partial unilateral ureteral obstruction leads to oxidative injury by relief of obstruction in both kidneys. The antioxidant allopurinol has a beneficial effect on renal MDA and GSH levels in both kidneys. PMID- 16351657 TI - Multifocal metastases of recurrent renal cell carcinoma successfully treated with a combination of low dose interleukin-2, alpha-interferon and radiotherapy. AB - A 59-year-old man presented with a 2-month history of left flank pain and a possibility of gross hematuria. Left renal cell carcinoma stage II was diagnosed and radical left nephrectomy was performed. Twenty-two months postoperatively, lung metastases were demonstrated and 6 x 10(6) units of alpha-interferon (IFN alpha) were administered for 9 months, only to keep the sizes of the metastases unchanged. Thirty-four months after the operation, liver metastases and bone metastasis in the left sacroiliac joint were revealed. The combination cytokine therapy was performed with 1.4 x 10(6) U of interleukin-2 (IL-2) and 3 x 10(6) U of IFN-alpha for 16 weeks, and the left sacroiliac joint metastasis was treated with radiation therapy of 4 Gy per day for 7 days. Six months after the 16 weeks of immunotherapy, computed tomography and bone scintigraphy revealed that the metastases of the lung, liver and bone substantially disappeared and this complete response is still kept after 16 months. PMID- 16351658 TI - Response of papillary renal cell carcinoma in a solitary kidney to high dose interleukin therapy. AB - Kidney cancer affects 36 000 Americans annually and is responsible for nearly 12 000 deaths every year in the US. Treatment with interleukin-2 (IL-2), the only FDA approved therapy for patients with advanced kidney cancer, is associated with a 10% complete response and a 12% partial response. To date, clear cell renal carcinoma has been the only histological type associated with response to IL-2 based therapy. In the current report, we describe a response to IL-2 therapy in a patient with type I papillary renal carcinoma. PMID- 16351659 TI - Massive epithelioid angiomyolipoma of the kidney in a young girl. AB - We report the case of a huge right renal tumor in a 17-year-old girl. Absence of fat on preoperative magnetic resonance imaging suggested renal cell carcinoma, and surgery was performed. Pathologic evaluation with HMB-45 immunohistochemical staining confirmed the diagnosis of epithelioid angiomyolipoma. The tumor consisted predominantly of epithelioid cells, and it could easily be misidentified as a renal cell carcinoma due to the paucity of the fat component. Previous reports have suggested that epithelioid angiomyolipomas have the potential to be malignant, and thus regular postoperative surveillance is recommended. Our patient had no signs of recurrence at her most recent follow up, 12 months after surgery. PMID- 16351660 TI - Renal matrix stones in an emphysematous pyelonephritis. AB - A 74-year-old-woman was referred to our hospital for further examination. Her chief complaint had been a high-grade fever, but she was seen at our hospital without fever. Plain fi lm of kidney, ureter and bladder drip infusion pyelography and abdominal computed tomography (CT) showed a gas shadow surrounding calcifications in the right renal pelvis. We diagnosed right emphysematous pyelonephritis. Since changes in the CT findings were not remarkable for 2 weeks, we performed laparoscopic right nephrectomy, even though her condition had not worsened. The renal pelvis contained brownish and clayish matter. We report here this rare case of renal matrix stones. PMID- 16351661 TI - Proteus syndrome with giant myelolipoma in the pelvis. AB - Proteus syndrome is a very rare and complex disorder with malformations and overgrowth of multiple tissues. This disorder was designated Proteus syndrome by Wiedemann et al. to denote its variable clinical expression. Our patient presented with macrodactyly, cerebriform appearance and a huge abdominal mass. A biopsy under laparotomy was performed, and histopathological examination revealed myelolipoma. Tumor resection was performed. To our knowledge, no case of Proteus syndrome presenting a myelolipoma in the retroperitoneal cavity has been reported before. PMID- 16351662 TI - Malignant phyllodes tumor of the prostate. AB - We report the case of a 47-year-old male patient who suffered from a malignant phyllodes tumor of the prostate with invasion to the rectum and urinary bladder. The local recurrence at the left scrotum was identified 6 years after radical cystoprostatectomy. Another 2 years after radical orchiectomy showed no evidence of secondary local recurrence or distant metastasis. Histopathologically, both primary and recurrent tumors showed an admixture of stromal and glandular components. However, while extensive squamous metaplasia was identified in the primary tumor, the recurrent tumor had only focal and mild squamous metaplasia. No dependable prognostic factor has been found to date. Here, we describe the morphological features and immunohistochemical presentations of malignant phyllodes tumor of the prostate and review the literature. PMID- 16351663 TI - Pure red cell aplasia in a prostate cancer patient treated with leuprolide acetate and chlormadinone acetate. AB - Pure red cell aplasia (PRCA) is characterized by anemia with reticulocytopenia but with normal leukocyte and platelet counts, and a bone marrow with a selective absence of erythroid precursor cells. Drug-induced PRCA is a rare secondary form of PRCA, and is usually acute and fully reversible by the withdrawal of the causative drugs. We report a rare case of PRCA in a prostate cancer patient treated with combined androgen blockade (CAB) consisted of leuprolide acetate as a luteinizing hormone-releasing hormone agonist and chlormadinone acetate as an antiandrogen. This case demonstrated that these drugs could be a cause of PRCA, and suggests that regular close monitoring for anemia is needed in prostate cancer patients treated with these drugs. PMID- 16351664 TI - Tumor lysis syndrome in a patient with metastatic, androgen independent prostate cancer. AB - Tumor lysis syndrome (TLS) is an uncommon, but well described, clinical entity that typically occurs following chemotherapy in patients with rapidly growing hematological malignancies. It is rarely described in patients with solid tumors. We report a case of TLS in a patient with metastatic adenocarcinoma of the prostate after treatment with paclitaxel chemotherapy. PMID- 16351665 TI - New strategy to ease the discomfort of insertion of nasogastric tubes. PMID- 16351666 TI - Reflections. PMID- 16351667 TI - Echocardiography for chest pain in the emergency department. PMID- 16351668 TI - Lipid-altering efficacy of switching from atorvastatin 10 mg/day to ezetimibe/simvastatin 10/20 mg/day compared to doubling the dose of atorvastatin in hypercholesterolaemic patients with atherosclerosis or coronary heart disease. AB - This randomised, double-blind study evaluated the efficacy and safety of ezetimibe/simvastatin (EZE/SIMVA) 10/20 mg tablet compared to doubling the atorvastatin (ATV) dose in hypercholesterolaemic patients with atherosclerotic or coronary heart disease (CHD). The study group included 435 male and female CHD patients (aged >or=18 years) who had not achieved their low-density lipoprotein cholesterol (LDL-C) goal of <2.50 mmol/l while on a stable dose of ATV 10 mg for >or=6 weeks. After a 1-week diet/stabilisation period, patients with LDL-C >or=2.50 mmol/l and 20% or a history of coronary heart or other atherosclerotic vascular disease were randomised to receive treatment with rosuvastatin 10 mg (n = 621), atorvastatin 10 mg (n = 189), simvastatin 20 mg (n = 194) or pravastatin 40 mg (n = 211) for 12 weeks. Significantly, more patients achieved 1998 and 2003 European LDL-C goals with rosuvastatin than with other statins after 12 weeks (p < 0.001). Rosuvastatin reduced LDL-C and total cholesterol levels significantly more than other statins, both in patients who were statin-naive and in patients who had received previous statin treatment (p < 0.05). All treatments were similarly well tolerated. In conclusion, greater reductions in LDL-C were achieved with rosuvastatin compared with atorvastatin, simvastatin and pravastatin, enabling more patients to achieve European LDL-C goals. PMID- 16351670 TI - Comparison of intravenous ibutilide vs. propafenone for rapid termination of recent onset atrial fibrillation. AB - This study was to evaluate the efficacy and safety of ibutilide and propafenone given intravenously in converting recent onset atrial fibrillation (AF). Eighty two consecutive patients with AF (onset in 2 h to 90 days) were randomly assigned to receive two 10-min infusions, 10 min apart, of either ibutilide (1 mg) or propafenone (70 mg). The treatment was considered successful if sinus rhythm occurred within 90 min after the beginning of infusion. Ibutilide had a significantly higher rate of cardioversion than propafenone (70.73 vs. 48.78%, p = 0.043). The patients with shorter AF duration or smaller left atrium diameter had a higher success rate. Nonsustained monomorphic ventricular tachycardia was the most serious adverse effect of ibutilide in 9.76% of patients, and hypotension and heart pause were the major serious adverse events in 17.07% of patients treated with propafenone. Ibutilide is more effective than intravenous propafenone for the cardioversion of recent onset AF, and the adverse effects are rare and transient. PMID- 16351671 TI - The lack of a simple relation between physician's percutaneous coronary intervention volume and outcomes in the era of coronary stenting: a two-centre experience. AB - The 2001 ACC/AHA guidelines recommend that percutaneous coronary intervention (PCI) operators perform at least 75 procedures per year to maintain their competency. We performed a post hoc analysis of prospectively gathered PCI data, in the current era of ubiquitous stent use, at two tertiary cardiac care centres. Operators were assigned to a low (<50 cases per year), intermediate (50-74 cases per year) or high volume (>or=75 cases per year) group. Complications evaluated were death, myocardial infarction, coronary perforation, emergent coronary artery bypass surgery and pericardial tamponade. Between 2000 and 2002, 51 operators performed 6,510 PCIs. Stents were used in 79% of cases. Major complications occurred in 0.45% (7/1,572 cases) for the low-volume group, 1.1% in the intermediate-volume group (16/1,438 cases) and 0.86% (30/3,500 cases) for the high-volume group. After adjusting for baseline factors, low- and intermediate volume operators were not significantly associated with major complications. This study questions the relationship between operator volume and PCI complications in the current era. PMID- 16351672 TI - The relationships between insulin resistance and components of metabolic syndrome in Taiwanese Asians. AB - Metabolic syndrome (MetS) is a complicated clinicopathological entity with clustering of cardiovascular and metabolic risk factors, which includes central obesity, hypertension, dyslipidemia and glucose intolerance. There were many studies investigating a wide variety of clinical and pathophysiological aspects of this syndrome. However, the cutoffs of the components of MetS are not yet being evaluated by measured the insulin resistance (IR) directly. In this study, we enrolled 564 (male/female: 250/314) middle-aged healthy subjects. Each of the male and the female group was further divided into four subgroups (group 1 to group 4). Group 4 had the top 25 percentile of most severe IR determined by insulin suppression test. We then obtain the mean values of each component of the MetS in group 4 and compared them with the definitions of World Health Organization, National Cholesterol Education Program Adult Treatment Panel III, European Study Group of Insulin Resistance and International Diabetes Federation. The means of the blood pressure (BP) (male, 125/81; female, 125/80 mmHg) and the triglyceride (TG) (male, 1.6; female, 1.4 mmol/l) in group 4 were lower, and the fasting plasma glucose (6.2 mmol/l) was higher than the cutoffs of the other four sets of the criteria. The means of the high-density lipoprotein cholesterol (male, 0.9; female, 1.03 mmol/l) and the body mass index (male, 26.9; female 26.1 kg/m(2)) in group 4 were consistent with the cutoffs of other four groups and also the Taiwan Health Department criteria. In conclusion, we suggest to lower the cutoffs of the BP from 140/90 to 125/80 mmHg, TG from 1.7 to 1.6 mmol/l for males and 1.4 mmol/l for females for MetS definition, at least in Taiwan. This may help to early detect subjects under high risk of future coronary heart disease and diabetes. Still, these newly proposed cutoffs need larger-scale epidemiological studies to confirm. PMID- 16351673 TI - Single vs. two steroid injections for carpal tunnel syndrome: a randomised clinical trial. AB - We investigated the efficacy of a single vs. double steroid injections in the treatment of carpal tunnel syndrome (CTS) in a randomised double-blind controlled trial. Patients with idiopathic CTS were randomised into (i) one group receiving a baseline methylprednisolone acetate injection plus a saline injection 8 weeks later and (ii) a second group receiving methylprednisolone acetate injection at baseline and at 8 weeks. The primary outcome was the Global Symptom Score (GSS). Forty patients were recruited. By 40 weeks, the mean GSS improved from 25.6 to 14.1 in the single-injection group whereas from 26.7 to 12.6 in the reinjection group, but there was no significant difference in GSS between the two groups (p = 0.26). There were also no significant differences in terms of electrophysiological and functional outcomes. The results suggest that an additional steroid injection confers no added benefit to a single injection in terms of symptom relief. PMID- 16351674 TI - Effectiveness of pre-emptive metoclopramide infusion in alleviating pain, discomfort and nausea associated with nasogastric tube insertion: a randomised, double-blind, placebo-controlled trial. AB - Aim of this study was to demonstrate that intravenous metoclopramide can reduce pain, nausea and discomfort during nasogastric tube (NGT) insertion in ED. This prospective, randomised, double-blind, placebo-controlled trial was conducted in the university-based ED. One-hundred patients were enrolled. Before NGT insertion, each eligible patient was randomised to one of the two treatment arms: one group received 2 cc of 10 mg IV metoclopramide, whereas others received 2 cc of normal saline. Before and after the procedure, pain, nausea and discomfort were evaluated using 100-mm visual analogue scale (VAS). This study was analysed using the paired sample test, the independent sample test and the chi(2) test. Forty-nine patients received metoclopramide, and 51 received normal saline. Although initial VAS levels elicited for pain, nausea and discomfort were similar, consequent VAS levels of those in the metoclopramide group were significantly lower as compared with those in the normal saline group. The mean differences of VAS levels were statistically significant for three symptoms (p < 0.001). Mean VAS levels of nausea, discomfort and pain during NGT insertion were significantly lower following administration of IV metoclopramide as compared with normal saline. PMID- 16351675 TI - Clinical effects of pharmacological variations in selective serotonin reuptake inhibitors: an overview. AB - Although the selective serotonin reuptake inhibitor (SSRI) class of antidepressants shares a common primary pharmacology, namely the inhibition of serotonin reuptake, their secondary pharmacology is remarkably heterogeneous. Inhibition of serotonin reuptake and the consequent increase in serotonin availability are responsible for the relief of depressive symptoms and for some of the adverse effects of this class of drugs. Transsynaptic effects such as modulation of signalling cascades, gene expression processes and neuroplasticity are also important in the mechanism of action of antidepressants. However, this review shows that secondary properties of the SSRIs may contribute to the differences in efficacy and tolerability between members of the class. For example, fluvoxamine has affinity for sigma(1)-receptors -- a property likely to be responsible for its particular efficacy in delusional depression. By understanding the properties of SSRIs and employing careful selection of agents for individual patients, physicians are more able to tailor antidepressant treatments to their patients' particular circumstances. PMID- 16351676 TI - Antidepressants and seizures: emphasis on newer agents and clinical implications. AB - Seizures are a known, relatively rare, consequence of antidepressant treatment. Risk estimates vary depending on the study, source of data and patient population, predisposed vs. nonpredisposed. For newer antidepressants (e.g. selective serotonin reuptake inhibitors, bupropion, mirtazepine, etc.), the risk is generally considered to be low (0.0%-0.4%) and not very different from the incidence of first seizure in the general population (0.07%-0.09%). Risk with tricyclic antidepressants at effective therapeutic doses is relatively high (0.4% to 1-2%). Seizure following overdose is a significant and relatively frequent event for some antidepressants. Patients being considered for antidepressant treatment should be screened for predisposition to seizures. Predisposed patients should receive antidepressants cautiously. The seizure potential of antidepressants in patients without a predisposition is low, especially for newer antidepressants. Seizure risk, along with other drug-related considerations, e.g. weight gain, sexual dysfunction and sedation, should be considered when prescribing an antidepressant. PMID- 16351677 TI - Evidence-based interventions to improve patient compliance with antihypertensive and lipid-lowering medications. AB - The MEDLINE database was searched from 1972 to June 2002 to identify studies of interventions designed to improve compliance with antihypertensive or lipid lowering medications. Studies were required to employ a controlled design, follow patients for >or=6 months and measure compliance by a method other than patient self-report. The literature review yielded 62 studies describing 79 interventions. Overall, 56% of interventions were reported to improve patient compliance. When only those studies meeting minimum criteria for methodological quality were considered, 22 interventions remained and 12 were recommended, because they demonstrated a significant improvement in compliance. Recommended interventions included fixed-dose combination drugs, once-daily or once-weekly dosing schedules, unit-dose packaging, educational counselling by telephone, case management by pharmacists, treatment in pharmacist- or nurse-operated disease management clinics, mailed refill reminders, self-monitoring, dose-tailoring, rewards and various combination strategies. Personalised, patient-focused programs that involved frequent contact with health professionals or a combination of interventions were the most effective at improving compliance. Less-intensive strategies, such as prescribing products that simplify the medication regimen or sending refill reminders, achieved smaller improvements in compliance but may be cost-effective due to their low cost. PMID- 16351678 TI - Are all non-thymidine analogue backbones appropriate for treating antiretroviral naive patients? AB - An increasing number of antiretroviral agents are available for the treatment of HIV infection. Many clinicians and patients prefer once-daily therapy, and this, in addition to accumulating evidence of toxicity associated with thymidine analogues, means many individuals commence a non-thymidine analogue-based regimen. Stavudine (d4T) is no longer recommended for initial therapy, and zidovudine (AZT) may also be associated with lipoatrophy. Despite investigations into nucleoside-sparing options, triple agent therapy with two nucleoside analogues [nucleoside reverse transcriptase inhibitors (NRTIs)] and a ritonavir boosted protease inhibitor (PI) or a non-nucleoside reverse transcriptase inhibitor (NNRTI) remains the mainstay. In this article, we review the advantages, and drawbacks, of different non-thymidine NRTI backbones. PMID- 16351679 TI - Thyroid cancer review 2: management of differentiated thyroid cancers. AB - This is the second review article on thyroid cancer dealing with the common differentiated types that comprise 90% of thyroid cancers. Despite clinical management guidelines offered by a number of organisations including the British Association of Otolaryngologists and Head and Neck Surgeon, The British Thyroid Association, The American Association Of Endocrinologists, The American Thyroid Association and the National Cancer Centre Network, debate regarding the management of differentiated thyroid cancer continues. This is because of the absence of good quality research evidence as a consequence of the natural history of this disease. PMID- 16351680 TI - Ezetimibe/simvastatin (INEGY) in the treatment of hyperlipidaemia. AB - Ezetimibe/simvastatin (INEGY), a dual inhibitor of both cholesterol production and absorption, is a new approach to the management of hyperlipidaemia. Recent studies have shown that it produces greater reductions in low-density lipoprotein (LDL) cholesterol than the single inhibition of statin therapy, enabling many more patients to achieve their LDL cholesterol treatment goals. With ezetimibe/simvastatin therapy, reductions of up to 61% from baseline have been seen in LDL cholesterol, with clear improvements in other associated lipid fractions. It has been well tolerated across all studies, with a safety profile similar to that of statin therapy. This article will review clinical experience to date with ezetimibe/simvastatin, commenting upon its place and potential value in the prevention of cardiovascular disease. PMID- 16351681 TI - Superimposition of nutcracker syndrome in a haematuric child with Henoch Schonlein purpura. AB - We report a child with Henoch-Schonlein purpura (HSP) who had shown persistent microscopic haematuria for 26 months and was found to have nutcracker syndrome by renal Doppler ultrasound. Haematuria subsided in accordance with radiological improvement of nutcracker syndrome. These findings suggest that the origin of urinary abnormalities in patients with HSP may not always be due to the renal involvement of vasculitis. PMID- 16351683 TI - Abdominal compartment syndrome: a new indication for operative intervention in severe acute pancreatitis. AB - The current management of severe acute pancreatitis (SAP) is maximal conservative therapy within an intensive care environment. The only commonly accepted indication for operative intervention is the presence of infected pancreatic necrosis. We present a case wherein a laparotomy performed for treatment of abdominal compartment syndrome (ACS) arising in the setting of SAP in the absence of pancreatic necrosis prevented early mortality and discuss the diagnosis and treatment of ACS as a new indication for operative intervention in SAP. PMID- 16351682 TI - Neglected peritonsillar abscess resulting in necrotizing soft tissue infection of the neck and chest wall. AB - Necrotizing soft tissue infection (NSTI) of the neck and chest wall resulting from neglected peritonsillar abscess is a relatively rare but highly lethal surgical condition which has received little attention in the literature. The case of a 54-year-old male patient affected by this unusual infection is reported. Our recent experience and literature data suggest that NSTI resulting from peritonsillar abscess is rapidly spreading and life threatening. High index of suspicion, early diagnosis, broad-spectrum antibiotics and aggressive surgical debridement are essential to its successful treatment. PMID- 16351684 TI - Premature ejaculation: presentation and associations. An audit of patients attending a sexual problems clinic. AB - The diagnostic register and case summaries of all male and female patients who attended the clinic on account of one or more sexual symptoms or relationship problems between 1 January 1992 and 31 December 1997 were reviewed. The index symptom or problem was considered the complaint that caused the patient most concern. When patients or couples were experiencing more than one sexual problem, the concomitant problems were tabulated against the index symptom. During this period, 1,056 (440 men and 616 women) patients with sexual symptoms were seen and 131 couples attended primarily with relationship problems. Overall, 18.2% of men referred to the clinic with sexual problems had premature ejaculation (PE), but this was the index symptom in only 11.6% of men. There was a high occurrence of PE in the partners of women presenting with sexual symptoms. Among couples presenting with relationship dissatisfaction or conflict, 18.3% of the male partners had ongoing PE. PMID- 16351685 TI - Idealhalers or realhalers? A comparison of Diskus and Turbuhaler. AB - Medication for the treatment of asthma and chronic obstructive pulmonary disease should be given locally by inhalation. There is, however, no such thing as an ideal inhaler, or 'Idealhaler', which has all desired properties with no drawbacks. In this short review, we have compared the relative merits of the two most commonly used dry powder inhalers -- Turbuhaler and Diskus. Clinical effect is related to the amount of inhaled drug that reaches the lungs, and this in turn depends on the amount of fine particles generated at inhalation. Turbuhaler is more than twice as effective as Diskus at generating fine particles, and the higher lung deposition with Turbuhaler is accompanied by a lower variability in lung deposition. Compared with Diskus, the lung deposition with Turbuhaler is affected less by factors such as humidity. PMID- 16351686 TI - How is the mouse germ-cell lineage established? AB - Six cells have been detected in the early mouse embryo that express the transcriptional repressor Blimp1--as also do all the 40 or so cells that constitute the founder germ cell pool a day later. Are these half-dozen cells the ancestors of the entire mouse germ cell lineage? PMID- 16351687 TI - Eggs forever? AB - A group of scientists from Harvard Medical School (Johnson et al., 2004) claims to have "established the existence of proliferative germ cells that sustain oocyte and follicle production in the postnatal mammalian ovary," expressing no doubts about their methods, results and conclusion. Johnson et al. based their conclusions of oocyte and follicular renewal from existing germline stem cells (GSC) in the postnatal mouse ovary on three types of observations: (1) A claimed discordance in follicle loss versus follicle atresia in the neonatal period and in the following pubertal and adult period; (2) immunohistochemical detection of proliferating GSC with meiotic capacity using combined markers for meiosis, germline, and mitosis; and (3) neo-folliculogenesis in ovarian chimeric grafting experiments with adult mice. Oogenesis is the process that transforms the proliferative oogonium into an oocyte through meiosis, followed by folliculogenesis and follicular and oocyte maturation. The most crucial part in producing a functional oocyte is firstly, initiation and completion of the first meiotic prophase, and secondly, enclosure of the resulting diplotene oocyte in a follicle. Neither of these two events has been shown to take place in Johnson et al.'s study of the postnatal mouse ovary. We hereby address the observations underpinning their hypothesis and conclude that it is premature to replace the paradigm that adult mammalian neo-oogenesis/folliculogenesis does not take place. PMID- 16351688 TI - Restoring stemness. AB - This essay is focused on a specific line of research toward regenerative therapies that is based on the use of embryonic stem cells but tries to avoid cloning techniques that are the heart of current ethical debates. PMID- 16351689 TI - Modulation of trophoblast stem cell and giant cell phenotypes: analyses using the Rcho-1 cell model. AB - Trophoblast giant cells are located at the maternal-embryonic interface and have fundamental roles in the invasive and endocrine phenotypes of the rodent placenta. In this report, we describe the experimental modulation of trophoblast stem cell and trophoblast giant cell phenotypes using the Rcho-1 trophoblast cell model. Rcho-1 trophoblast cells can be manipulated to proliferate or differentiate into trophoblast giant cells. Differentiated Rcho-1 trophoblast cells are invasive and possess an endocrine phenotype, including the production of members of the prolactin (PRL) family. Dimethyl sulfoxide (DMSO), a known differentiation-inducing agent, was found to possess profound effects on the in vitro development of trophoblast cells. Exposure to DMSO, at non-toxic concentrations, inhibited trophoblast giant cell differentiation in a dose dependent manner. These concentrations of DMSO did not significantly affect trophoblast cell proliferation or survival. Trophoblast cells exposed to DMSO exhibited an altered morphology; they were clustered in tightly packed colonies. Trophoblast giant cell formation was disrupted, as was the expression of members of the PRL gene family. The effects of DMSO were reversible. Removal of DMSO resulted in the formation of trophoblast giant cells and expression of the PRL gene family. The phenotype of the DMSO-treated cells was further determined by examining the expression of a battery of genes characteristic of trophoblast stem cells and differentiated trophoblast cell lineages. DMSO treatment had a striking stimulatory effect on eomesodermin expression and a reciprocal inhibitory effect on Hand1 expression. In summary, DMSO reversibly inhibits trophoblast differentiation and induces a quiescent state, which mimics some but not all aspects of the trophoblast stem cell phenotype. PMID- 16351690 TI - Stem/progenitor and intermediate cell types and the origin of human prostate cancer. AB - Theories of cell lineage in human prostatic epithelium, based on protein expression, propose that basal and luminal cells: 1) are either independently capable of self-renewal or 2) arise from stem cells expressing a full spectrum of proteins (p63, cytokeratins CK5/14, CK8/18, and glutathione-S-transferase-pi [GST pi]) similar to cells of the embryonic urogenital sinus (UGS). Such embryonic like stem cells are thought to give rise to mature basal cells and secretory luminal cells. By single cell cloning of an immortalized, normal human prostate derived, non-tumorigenic RWPE-1 cell line, we isolated and characterized two epithelial cell types, WPE-stem and WPE-int. WPE-stem cells show: i) strong, sixfold greater nuclear expression of p63; ii) nearly twofold greater expression of CK14; iii) threefold less CK18, and iv) low androgen receptor (AR) expression as compared with WPE-int cells. WPE-stem cells are androgen-independent for growth and survival. WPE-int cells express very low p63 and CK5/14, and high CK18. WPE-int cells are androgen-independent for growth and survival but are highly responsive as shown by androgen induction of AR and prostate specific antigen (PSA). Compared with WPE-int cells, WPE-stem cells are smaller and show more rapid growth. WPE-stem cells can grow in an anchorage-independent manner in agar with 4.5-fold greater cloning efficiency and as free floating "prostaspheres" in liquid medium; and express over 40-fold higher matrix metalloproteinase-2 activity. These results indicate that WPE-stem cells express several features characteristic of stem/progenitor cells present in the UGS and in adult prostatic epithelium. In contrast, WPE-int cells have an intermediate, committed phenotype on the pathway to luminal cell differentiation. We propose that in normal prostatic epithelium, cells exist at many stages in a continuum of differentiation progressing from stem cells to definitive basal and luminal cells. Establishment and characterization of clones of human prostatic epithelial cells provide novel models for determining cell lineages, the origin of prostate cancer, and for developing new strategies for tumor prevention and treatment. PMID- 16351691 TI - Human fibroblast-derived cell lines have characteristics of embryonic stem cells and cells of neuro-ectodermal origin. AB - Fibroblasts are the most ubiquitous cells in complex organisms. They are the main cells of stromal tissue and play an important role in repair and healing of damaged organs. Here we report new data-initially serendipitous findings-that fibroblast-derived cell line (human fetal lung derived cells, MRC-5) have the morphology, growth rate and gene expression pattern characteristic of embryonic stem cells and cells of neuro-ectodermal origin. We have developed a serum-free culture system to maintain these cells in proliferative state. We discovered that, at proliferative state, these cells express transcription factors of pluripotent cells, OCT-3/4 and REX-1, and embryonic cell surface antigens SSEA-1, SSEA-3, and SSEA-4, as well as TRA-1-60 and TRA-1-81. In addition to embryonic cell markers, the fibroblasts expressed neuroectodermal genes: Musashi-1, nestin, medium neurofilament, and beta-III tubulin. RT-PCR data revealed that mesencephalic transcription factors, Nurr-1 and PTX-3, were also expressed in MRC 5 cells, and that these cells could be induced to express tyrosine hydroxylase (TH). Expression of TH followed down-regulation of genes associated with cell proliferation, OCT-3/4, REX-1, and beta-catenin. These data indicate that the cells commonly known as fibroblasts have some of the characteristics of stem cells, and can be induced to become neuroectodermal cells and perhaps even mature neurons. PMID- 16351692 TI - Fate of embryonal carcinoma cells injected into postimplantation mouse embryos. AB - Embryonal carcinoma (EC) cells, stem cells of teratocarcinoma, represent an excellent model to study the developmental mechanisms that, inappropriately reactivated, can drive tumorigenesis. EC cells are very aggressive, and grow rapidly when injected into adult syngeneic mice. However, when injected into blastocysts, they revert to normality, giving rise to chimeric animals. In order to study the ability of postimplantation embryonic environment to "normalize" tumorigenic cells, and to study their homing, we transplanted F9, Nulli-SCC1, and P19 EC cells into 8 to 15-day allogenic CD1 mouse embryos, into allogenic CD1 newborns, and into syngeneic adult mice, and evaluated tumor formation, spreading, and homing. We found that, although at all embryonic stages successful transplantation occurred, the chances of developing tumors after birth increased with the time of injection of EC cells into the embryo. In addition, using enhanced green fluorescent protein-expressing F9 cells, we demonstrated that the cells not giving rise to tumors remained latent and could be tracked down in tissues during adulthood. Our data indicate that the embryonic environment retains a certain ability to "normalize" tumor cells also during post implantation development. This could occur through yet unknown epigenetic signals triggering EC cells' differentiation. PMID- 16351693 TI - A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China. AB - Immunological methods have been developed for the diagnosis of Myxobolus rotundus but their use has been limited for the prevention and therapy of this serious parasitic pathogen. Phage display antibody libraries are a powerful technique for the development of antibodies to molecules of interest and have advantages over traditional hybridroma approaches. In the present study, four antigen fractions related to M. rotundus were prepared and a combined phage display single-chain antibody fragments (ScFv) library was constructed against this parasite. Preliminary analysis indicated that a combined antibody library of about 2.08 x 10(5) individual clones and high diversity was generated. After four rounds of screening (bio-panning) against soluble spore protein prepared from lysed, intact, mature M. rotundus spores, a strain monoclonal phage display ScFv, termed pCAN-6H9, with better affinity, was isolated. The pCAN-6H9 gene fragment was sequenced and analysed. The specificity of pCAN-6H9 was further demonstrated by dot-blot. In competition enzyme-linked immunosorbent assay, both the original and enriched phage-displayed ScFv repertoire showed significant inhibition of mouse anti-M. rotundus serum binding to coated antigen, while the inhibition rate of monoclonal pCAN-6H9 phage particles was only 11.83%. PMID- 16351694 TI - Efficacy of injection vaccines against Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum). AB - Efficacy of mineral oil-based experimental injection vaccines against Flavobacterium psychrophilum were tested in rainbow trout, Oncorhynchus mykiss (Walbaum), under laboratory and field conditions. The vaccines consisted of formalin- or heat-inactivated whole bacterium cell preparations of two different serotypes (Fd and Th) or a combination of serologically different F. psychrophilum (Fd and/or Th and/or Fp(T);Th). Specific antibody responses against the bacterium in plasma and skin mucus were evaluated post-vaccination with enzyme-linked immunosorbent assay. Efficacy of the vaccinations was determined by challenge trials to F. psychrophilum with the vaccinated rainbow trout. Significantly higher antibody levels in plasma were detected in vaccinated fish compared with mock-vaccinated fish. Injection vaccination did not trigger specific antibody production in the skin mucus. Significantly higher survival of i.p. vaccinated fish compared with non-vaccinated fish was observed during the challenge. The results suggest that mineral oil-based injectable vaccines containing formalin- or heat-inactivated virulent cells of F. psychrophilum effectively triggered specific antibody production and protected the fish against bacterial cold water disease. PMID- 16351695 TI - Atypical growth of Renibacterium salmoninarum in subclinical infections. AB - Two growth types of Renibacterium salmoninarum were isolated from subclinically infected rainbow trout, one producing the smooth colonies typical of R. salmoninarum and the other forming a thin film on the surface of the agar with no separate colonies. The atypical growth was present on kidney disease medium agar in primary cultures of the kidney but not on selective kidney disease medium (SKDM). Fluorescent antibody staining of the fresh isolate and polymerase chain reaction amplification were the most reliable techniques to identify the atypical growth of R. salmoninarum. The condition was reversible, with growth reverting from atypical to the smooth colony form in experimentally infected rainbow trout and under laboratory conditions. There was no mortality, or any clinical signs of bacterial kidney disease (BKD) in the fish challenged with the atypical growth, although small numbers of smooth colonies of R. salmoninarum were isolated from 8% of these fish. The atypical growth reported here may explain some of the failures of culture, when SKDM agar alone is used for the detection of BKD in subclinically infected fish. PMID- 16351696 TI - Infectious pancreatic necrosis virus in Atlantic salmon, Salmo salar L., post smolts in the Shetland Isles, Scotland: virus identification, histopathology, immunohistochemistry and genetic comparison with Scottish mainland isolates. AB - During mid-June 1999 peak mortalities of 11% of the total stock per week were seen at a sea cage site of Atlantic salmon, Salmo salar L., post-smolts in the Shetland Isles, Scotland. Virus was isolated on chinook salmon embryo (CHSE) cells in a standard diagnostic test and infectious pancreatic necrosis virus (IPNV) identified by enzyme-linked immunosorbent assay. IPNV was confirmed as serogroup A by a cell immunofluorescent antibody test using the cross-reactive monoclonal antibody AS-1. Four weeks after the main outbreak, virus titres in surviving moribund fish were assayed at >10(10) TCID50 g(-1) kidney. Histopathology of moribund fish was characterized by pancreatic acinar cell necrosis and a marked catarrhal enteritis of the intestinal mucosa. In the liver, necrosis, leucocytic infiltration and a generalized cell vacuolation were noted. IPNV-specific immunostaining was demonstrated in pancreas, liver, heart, gill and kidney tissue. The nucleotide sequence of the coding region of segment A was determined from the Shetland isolate. A 1180 bp fragment of the VP2 gene of this isolate was compared with a 1979 reference isolate from mainland Scottish Atlantic salmon, La/79 and another more recent mainland isolate, 432/00. Both A2 isolates were derived from carrier fish without signs of IPN and serotyped by a plaque neutralization test. The Shetland isolate shows a different nucleotide and amino acid sequence compared with the two isolates from carrier fish. These latter isolates showed identical amino acid sequences in the fragment examined, despite the 21 years separating the isolations. Sequence comparisons with other A2 (Sp) isolates on the database confirm all three Scottish isolates are A2 (Sp). PMID- 16351697 TI - In infectious pancreatic necrosis virus carrier Atlantic salmon, Salmo salar L., post-smolts, almost all kidney macrophages ex vivo contain a low level of non replicating virus. AB - The level of infection by infectious pancreatic necrosis virus (IPNV) of kidney macrophages from 12 asymptomatic carrier Atlantic salmon post-smolts was studied. Kidney leucocytes were fractionated on 34/51% Percoll gradients, allowed to adhere to plastic wells overnight, washed to remove non-adherent cells and cultured for up to 7 days with or without renewal of medium on day 3. On day 1, supernatants were harvested, macrophages were counted, lysed and IPNV in the supernatants and lysates was titred in chinook salmon embryo (CHSE-214) cells. The multiplicity of infection ranged between 1:2.2 and 1:7.4 (virus:macrophages). On day 3, the titres of IPNV in macrophage lysates decreased and in wells where the medium was renewed on day 3, IPNV was no longer detectable on day 7. In the supernatants, one fish was positive for IPNV on day 1, four fish on day 3 but none were detectably positive on day 7. In parallel wells in which the medium was not renewed, on day 7 IPNV was detected in macrophage lysates of three fish and the supernatants were also IPNV positive in two of these fish. This suggests that virus might be shed from infected macrophages and then reinfect other macrophages. When macrophages were serially diluted in wells and cultured for 24 h, IPNV could be cultured from macrophage lysates of wells containing between two and 70 macrophages. These results indicate that a very high proportion of the adherent kidney macrophages must be infected with very few non-replicating virions. PMID- 16351698 TI - Spinal deformity in a sandtiger shark, Carcharias taurus Rafinesque: a clinical pathological study. AB - The present study investigated spinal deformity in a captive sandtiger shark and describes the clinical and histopathological features of the lesion. Clinically, the shark presented a marked spinal curvature in the region between the pectoral girdle and cranial dorsal fin. Radiographic studies revealed scoliosis and kyphosis of the vertebral column with a modification of the basic structure of five vertebrae. Post-mortem examination confirmed the results of radiographic studies and histological examination showed remodelling and cartilaginous proliferation of the vertebral bodies together with necrosis, haemorrhage and signs of a local chronic inflammatory response extending to the surrounding muscle. Different aetiologies are proposed and discussed but nutritional imbalance (dietary vitamin deficiency) and/or traumatic injury are considered the most likely contributing factors. PMID- 16351699 TI - Steatitis, fin loss and skin ulcers of channel catfish, Ictalurus punctatus (Rafinesque), fingerlings fed salmonid diets. PMID- 16351700 TI - Haste makes waist... PMID- 16351701 TI - Clothing size as an indicator of adiposity, ischaemic heart disease and cardiovascular risks. AB - OBJECTIVE: To relate subjects' clothing sizes to waist circumference, body mass index (BMI) and to the risks of ischaemic heart disease, hypertension and diabetes mellitus, and to derive cut-off levels of clothing size that correspond to increased health risks. DESIGN: Cross-sectional study. Setting Glasgow Royal Infirmary. PARTICIPANTS: A stratified subsample of 201 men and 161 women aged 27 67 years from the Glasgow monitoring coronary (MONICA) risk factor survey. MAIN OUTCOME MEASURES: Measured waist, weight and height, blood pressure and history of ischaemic heart disease (angina, myocardial infarction or angioplasty), hypertension and diabetes mellitus, and medications. RESULTS: There were 15.5% of men and 11.2% of women with ischaemic heart disease, 14.9% of men and 12.4% of women with hypertension, and 4.5% of men and 3.1% of women with diabetes mellitus. Age and smoking adjusted prevalences of these cardiovascular risks, as well as increased adiposity (waist circumference > or =102 cm in men, > or =88 cm in women or BMI > or = 30 kg m(-2)) rose with increasing clothing size. Clothing size correlated (P < 0.001) linearly with indices of adiposity. Clothing sizes which correspond to waist circumference action level 1 (94 cm in men, 80 cm in women) and action level 2 (102 cm in men, 88 cm in women) or to standardized BMI cut-offs (25 and 30 kg m(-2)) were estimated. Height has minimal influences on clothing size. Compared with men with trouser waist below 36 inches or women with UK dress size below 16, the age and smoking adjusted odds ratios for the risk of having at least one of the major health problems (ischaemic heart disease, hypertension or diabetes mellitus) were 3.9 (95% CI: 1.8-8.3) in men and 7.0 (95% CI: 2.5-19.4) in women who had trouser size > or =38 inches or UK dress size > or =18, respectively. CONCLUSIONS: The present study showed that men and women with large clothing size are at increased risk of ischaemic heart disease, hypertension and diabetes mellitus. Men's trouser size equal or larger than 38 in the UK and USA or 97 in Europe and women's dress size equal or larger than 18 in the UK or 16 in the USA or 48 in Europe could be used to promote self-awareness of increased health risks by the general public. PMID- 16351702 TI - Determinants of fruit and vegetable consumption among 6-12-year-old children and effective interventions to increase consumption. AB - PURPOSE: To review the current literature about potential determinants of fruit and vegetable intakes and effective intervention strategies to increase the consumption of fruits and vegetables among 6-12-year-old children. METHODS: A structured review of literature located in PubMed and Psychinfo electronic literature databases. RESULTS: Of all determinants, the availability and accessibility of fruit and vegetables and taste preferences were most consistently and most positively related to consumption. There was some evidence that parental fruit and vegetable intakes, knowledge of intake recommendations and skills had a positive association with children's intakes, whereas television viewing, exposure to television advertisement, and having a snack bar at school were associated with lower intakes of fruit and vegetables. Multi-component school-based interventions that combined classroom curriculum, parent and food service components showed the greatest promise for fruit and vegetable promotion among children. School fruit and vegetable subscription programmes, scout-based interventions, and fruit and vegetables education via computer multi-media channels also appear promising. CONCLUSIONS: Interventions should improve the availability and accessibility of fruit and vegetables to children, and should aim to improve their taste preferences for them. Such interventions should be of a multi-component nature, school-based or use other social channels and may include multi-media channels. PMID- 16351703 TI - Dietary problems in patients with severe chronic obstructive pulmonary disease. AB - BACKGROUND AND AIMS: Low body weight and low fat-free mass-index (FFMI) are common in patients with severe chronic obstructive pulmonary disease (COPD). Several factors seem to contribute. The aims of the present observational study were to investigate dietary problems in patients with severe COPD and to compare dietary problems to nutritional status, energy intake and smoking habits. METHODS: Nutritional status was assessed in 73 stable outpatients using body mass index and FFMI by single-frequency bioelectrical impedance. Lung function, smoking habits, energy intake and dietary problems were also assessed. RESULTS: The most frequently reported dietary problems were 'anorexia', 'dyspeptic symptoms other than diarrhoea', 'slimming', 'fear of gaining weight', 'dyspnoea', 'diarrhoea', 'depression, anxiety, solitude'. Smoking habits and gender had impact on the kind of dietary problems reported. Reporting two dietary problems correlated to low FFMI, whereas reporting one or more dietary problems correlated to decreased energy intake. CONCLUSION: Dietary problems are common in the group studied and related to smoking habits and gender. Dietary problems affect energy intake and FFMI negatively. It is important to recognize dietary problems and to offer intervention of the dietary problems as a part of the dietary intervention. PMID- 16351704 TI - Older women and dietary advice: occurrence, comprehension and compliance. AB - BACKGROUND: When a person becomes ill, traditional food habits may come into conflict with the disease-related recommended diet. AIM: The aim was to study perceptions of receiving dietary advice, the occurrence and comprehension of such advice and compliance among older women diagnosed with Parkinson's disease, stroke or rheumatoid arthritis. METHODS: The study took an ethnographic approach. Fifty-four community-dwelling women, 64-88 years of age, were interviewed using an open-ended interview technique during visits to their homes. FINDINGS: Two themes were found: 'dietary advice - occurrence and comprehension' describes whether the women had received dietary advice and, if so, how they understood the information. In the theme 'compliance with dietary advice', two principal reasons for complying or not complying with advice were found: First, women expressed a 'food interest', such that they were either 'interested in disease-related diet' or held a general view of the significance of foods and complied with the advice for their own 'health interest'. Secondly, the women were 'uninterested in food changes'. This could be because of 'poor appetite', 'food and disease ambivalence', 'habitual and preferred foods'. CONCLUSION: Dietary advice should be based on women's food preferences and habitual foods. It is important to inform about known relations between food and disease, but also to support eating favourite foods, thereby facilitating women's well-being. PMID- 16351705 TI - Interprofessional education in undergraduate healthcare programmes: the reaction of student dietitians. AB - BACKGROUND: Interprofessional education (IPE) is a novel teaching and learning initiative where students of more than one health profession learn interactively together. However, despite its potential for improving interprofessional relationships, there is little information regarding the participation of student dietitians in IPE. The aim of this paper was to consider the reaction of student dietitians to an IPE course in order to stimulate debate between dietitians regarding the issues relating to IPE. METHODS: Student dietitians participated in an IPE course consisting of seven sessions on communication and ethics in health care together with students of medicine and nursing. Student dietitians completed an evaluation questionnaire following each session that surveyed their reaction to the session using both a Likert scale and free-text comments. RESULTS: Twenty six student dietitians completed the IPE course. All sessions were rated positively for interest value (P < or = 0.14), learning experience (P < or = 0.036) and value for clinical practice (P < or = 0.05). The limited number of free-text comments indicated some positive experiences regarding interprofessional learning, teaching content and teaching strategy. CONCLUSIONS: This is one of very few evaluations to describe the reaction of student dietitians to IPE. Student dietitians had largely positive reactions to the IPE course. Further research is required to evaluate whether these positive reactions were a direct consequence of the inclusion of students from other health professions and whether these translate into positive effects on learning, behaviour and results. The opportunities for the dietetic profession posed by students' involvement in IPE are discussed. PMID- 16351706 TI - Empathy, empowerment, political will and a coeliac specialist dietitian in every hospital? PMID- 16351709 TI - p21WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells. AB - BACKGROUND: p21WAF1, implicated in the cell cycle control of both normal and malignant cells, can be induced by p53-dependent and independent mechanisms. In some cells, MEKs/ERKs regulate p21WAF1 transcriptionally, while in others they also affect the post-transcriptional processes. In myogenic differentiation, p21WAF1 expression is also controlled by the myogenic transcription factor MyoD. We have previously demonstrated that the embryonal rhabdomyosarcoma cell line undergoes growth arrest and myogenic differentiation following treatments with TPA and the MEK inhibitor U0126, which respectively activate and inhibit the ERK pathway. In this paper we attempt to clarify the mechanism of ERK-mediated and ERK-independent growth arrest and myogenic differentiation of embryonal and alveolar rhabdomyosarcoma cell lines, particularly as regards the expression of the cell cycle inhibitor p21WAF1. RESULTS: p21WAF1 expression and growth arrest are induced in both embryonal (RD) and alveolar (RH30) rhabdomyosarcoma cell lines following TPA or MEK/ERK inhibitor (U0126) treatments, whereas myogenic differentiation is induced in RD cells alone. Furthermore, the TPA-mediated post transcriptional mechanism of p21WAF1-enhanced expression in RD cells is due to activation of the MEK/ERK pathway, as shown by transfections with constitutively active MEK1 or MEK2, which induces p21WAF1 expression, and with ERK1 and ERK2 siRNA, which prevents p21WAF1 expression. By contrast, U0126-mediated p21WAF1 expression is controlled transcriptionally by the p38 pathway. Similarly, myogenin and MyoD expression is induced both by U0126 and TPA and is prevented by p38 inhibition. Although MyoD and myogenin depletion by siRNA prevents U0126 mediated p21WAF1 expression, the over-expression of these two transcription factors is insufficient to induce p21WAF1. These data suggest that the transcriptional mechanism of p21WAF1 expression in RD cells is rescued when MEK/ERK inhibition relieves the functions of myogenic transcription factors. Notably, the forced expression of p21WAF1 in RD cells causes growth arrest and the reversion of anchorage-independent growth. CONCLUSION: Our data provide evidence of the key role played by the MEK/ERK pathway in the growth arrest of Rhabdomyosarcoma cells. The results of this study suggest that the targeting of MEK/ERKs to rescue p21WAF1 expression and myogenic transcription factor functions leads to the reversal of the Rhabdomyosarcoma phenotype. PMID- 16351710 TI - Simplified screening for the detection of soluble fusion constructs expressed in E. coli using a modular set of vectors. AB - BACKGROUND: The solubility of recombinant proteins expressed in bacteria is often disappointingly low. Several strategies have been developed to improve the yield and one of the most common strategies is the fusion of the target protein with a suitable partner. Despite several reports on the successful use of each of these carriers to increase the solubility of some recombinant proteins, none of them was always successful and a combinatorial approach seems more efficient to identify the optimal combination for a specific protein. Therefore, the efficiency of an expression system critically depends on the speed in the identification of the optimal combination for the suitable fusion candidate in a screening process. This paper describes a set of expression vectors (pETM) designed for rapid subcloning, expression and subsequent purification using immobilized metal affinity chromatography (IMAC). RESULTS: A single PCR product of two Yellow Fluorescent Proteins (EYFPs) was cloned into 18 vectors comprising identical restriction sites and varying fusion partners as well as differing protease recognition sites. After a small-scale expression, the yields of the different constructs were compared using a Coomassie stained SDS-polyacrylamide gel and the results of this preliminary screening were then confirmed by large scale purification. The yields were calculated and the stability of the different constructs determined using three independent conditions. The results indicated a significant correlation between the length and composition of non-native amino acid tails and stability. Furthermore, the buffer specificity of TEV and 3C proteases was tested using fusion proteins differing only in their protease recognition sequence, and a His-GST-EYFP construct was employed to compare the efficiency of the two alternative affinity purification methods. CONCLUSION: The experiments showed that the set of pETM vectors could be used for the rapid production of a large array of different constructs with specific yield, stability, and cleavage features. Their comparison allowed the identification of the optimal constructs to use for the large-scale expression. We expect that the approach outlined in this paper, i.e. the possibility to obtain in parallel fusion products of the target protein with different partners for a preliminary evaluation, would be highly beneficial for all them who are interested in the rapid identification of the optimal conditions for protein expression. PMID- 16351711 TI - A microarray data-based semi-kinetic method for predicting quantitative dynamics of genetic networks. AB - BACKGROUND: Elucidating the dynamic behaviour of genetic regulatory networks is one of the most significant challenges in systems biology. However, conventional quantitative predictions have been limited to small networks because publicly available transcriptome data has not been extensively applied to dynamic simulation. RESULTS: We present a microarray data-based semi-kinetic (MASK) method which facilitates the prediction of regulatory dynamics of genetic networks composed of recurrently appearing network motifs with reasonable accuracy. The MASK method allows the determination of model parameters representing the contribution of regulators to transcription rate from time series microarray data. Using a virtual regulatory network and a Saccharomyces cerevisiae ribosomal protein gene module, we confirmed that a MASK model can predict expression profiles for various conditions as accurately as a conventional kinetic model. CONCLUSION: We have demonstrated the MASK method for the construction of dynamic simulation models of genetic networks from time series microarray data, initial mRNA copy number and first-order degradation constants of mRNA. The quantitative accuracy of the MASK models has been confirmed, and the results indicated that this method enables the prediction of quantitative dynamics in genetic networks composed of commonly used network motifs, which cover considerable fraction of the whole network. PMID- 16351712 TI - Prediction of MHC class II binding peptides based on an iterative learning model. AB - BACKGROUND: Prediction of the binding ability of antigen peptides to major histocompatibility complex (MHC) class II molecules is important in vaccine development. The variable length of each binding peptide complicates this prediction. Motivated by a text mining model designed for building a classifier from labeled and unlabeled examples, we have developed an iterative supervised learning model for the prediction of MHC class II binding peptides. RESULTS: A linear programming (LP) model was employed for the learning task at each iteration, since it is fast and can re-optimize the previous classifier when the training sets are altered. The performance of the new model has been evaluated with benchmark datasets. The outcome demonstrates that the model achieves an accuracy of prediction that is competitive compared to the advanced predictors (the Gibbs sampler and TEPITOPE). The average areas under the ROC curve obtained from one variant of our model are 0.753 and 0.715 for the original and homology reduced benchmark sets, respectively. The corresponding values are respectively 0.744 and 0.673 for the Gibbs sampler and 0.702 and 0.667 for TEPITOPE. CONCLUSION: The iterative learning procedure appears to be effective in prediction of MHC class II binders. It offers an alternative approach to this important prediction problem. PMID- 16351713 TI - The host response to the probiotic Escherichia coli strain Nissle 1917: specific up-regulation of the proinflammatory chemokine MCP-1. AB - BACKGROUND: The use of live microorganisms to influence positively the course of intestinal disorders such as infectious diarrhea or chronic inflammatory conditions has recently gained increasing interest as a therapeutic alternative. In vitro and in vivo investigations have demonstrated that probiotic-host eukaryotic cell interactions evoke a large number of responses potentially responsible for the effects of probiotics. The aim of this study was to improve our understanding of the E. coli Nissle 1917-host interaction by analyzing the gene expression pattern initiated by this probiotic in human intestinal epithelial cells. METHODS: Gene expression profiles of Caco-2 cells treated with E. coli Nissle 1917 were analyzed with microarrays. A second human intestinal cell line and also pieces of small intestine from BALB/c mice were used to confirm regulatory data of selected genes by real-time RT-PCR and cytometric bead array (CBA) to detect secretion of corresponding proteins. RESULTS: Whole genome expression analysis revealed 126 genes specifically regulated after treatment of confluent Caco-2 cells with E. coli Nissle 1917. Among others, expression of genes encoding the proinflammatory molecules monocyte chemoattractant protein-1 ligand 2 (MCP-1), macrophage inflammatory protein-2 alpha (MIP-2alpha) and macrophage inflammatory protein-2 beta (MIP-2beta) was increased up to 10 fold. Caco-2 cells cocultured with E. coli Nissle 1917 also secreted high amounts of MCP-1 protein. Elevated levels of MCP-1 and MIP-2alpha mRNA could be confirmed with Lovo cells. MCP-1 gene expression was also up-regulated in mouse intestinal tissue. CONCLUSION: Thus, probiotic E. coli Nissle 1917 specifically upregulates expression of proinflammatory genes and proteins in human and mouse intestinal epithelial cells. PMID- 16351714 TI - Primary healthcare provision and Chronic Fatigue Syndrome: a survey of patients' and General Practitioners' beliefs. AB - BACKGROUND: The current study was conducted as part of a research project into the evaluation and assessment of healthcare provision and education in Chronic Fatigue Syndrome (CFS). One aim of the study was the development of informative and educational literature for both General Practitioners (GP) and sufferers. Issues such as diagnosis, management and treatment of the syndrome should be included in information booklets written by healthcare professionals. It was important to begin the process by assessing the level of specialist knowledge that existed in typical GP surgeries. This data would then be compared to data from CFS patients. METHOD: 197 survey booklets were sent to CFS sufferers from an existing research panel. The patients approached for the purpose of the study had been recruited onto the panel following diagnosis of their illness at a specialised CFS outpatient clinic in South Wales. A further 120 booklets were sent to GP surgeries in the Gwent Health Authority region in Wales. RESULTS: Results from the study indicate that the level of specialist knowledge of CFS in primary care remains low. Only half the GP respondents believed that the condition actually exists. CONCLUSION: Steps are recommended to increase the knowledge base by compiling helpful and informative material for GPs and patient groups. PMID- 16351715 TI - Recognition of delirium in ICU patients: a diagnostic study of the NEECHAM confusion scale in ICU patients. AB - BACKGROUND: A delirium, is a serious, high-frequency complication in intensive care unit (ICU) patients. The consequences of this complication range from high morbidity and mortality to greater need for nursing care. Despite these, delirium is often not recognized and there for not treated. In this study a nursing screening instrument, the NEECHAM confusion scale, was studied for early recognition of delirium ICU patients. This scale proved valid and reliable in several studies in the general hospital population. METHODS: In this study validity and reliability were tested in a prospective cohort of 105 patients. Gold standard for delirium was an independent DSM-IV diagnosis. User friendliness was tested by structured evaluation of nurses' experiences working with the scale. RESULTS: The NEECHAM confusion scale showed high internal consistency (Cronbach's alpha 0.88) and an interrater reliability of Cohen's Kappa 0.60. The concurrent validity with the DSM-IVcriteria showed a strong link (chi-square 67.52, p [less than or equal to] 0.001). Sensitivity was high, 97% and specificity was good 83%. ICU nurses completed the NEECHAM confusion rating in 3.69, +/- 1.21 minutes average. In general the nurses were positive about the NEECHAM confusion scale. They were able to collect data during regular care, but experienced problems in rating the scale in intubated patients. The items in themselves were clear, the content validity, measured by the language used was rated good. CONCLUSION: The psychometric characteristics of the NEECHAM confusion scale of this ICU study are generally consistent with validity research previously reported for the general hospital population. The psychometric characteristics and the ease of use of the NEECHAM confusion scale enables ICU nurses to early recognize delirium. Further study, especially in intubed patients is recommended. PMID- 16351716 TI - Vitamin status and cognitive function in a long-term care population. AB - BACKGROUND: Ageing can be associated with poor dietary intake, reduced nutrient absorption, and less efficient utilization of nutrients. Loss of memory and related cognitive function are also common among older persons. This study aimed to measure the prevalence of inadequate vitamin status among long-term care patients and determine if an association exists between vitamin status and each of three variables; cognitive function, vitamin supplementation, and medications which alter gastric acid levels. METHODS: Seventy-five patients in a long-term care hospital in Guelph, Ontario were recruited to a cross-sectional study. 47 were female and the mean age was 80.7 (+/-11.5) years, ranging from 48 to 100 years. Blood was used to measure levels of vitamins B12 (cobalamin), B6 (pyridoxal-5'-phosphate/PLP), erythrocyte folate, vitamin B3 (niacin) and homocysteine (Hcy). The Standardized Mini-Mental State Examination (SMMSE) was administered to measure cognitive function. A list of medications and vitamin supplementation for each patient was provided by the pharmacy. RESULTS: The prevalence of low vitamin (B12, B6, erythrocyte folate, niacin) or high metabolite (homocysteine) levels among 75 patients were as follows: B12 <148 pmol/L in 5/75 (6.7%); B12 between 148 and 221 pmol/L in 26/75 (34.7%); B6 13.3 micromol/L in 31/75 (41.3%). There was no significant difference among residents grouped into marked (n = 44), mild (n = 14), or normal (n = 9) cognitive function when evaluating the effect of vitamin status. There were no significant differences in mean B12 and homocysteine levels between users and non-users of drug therapy (Losec, Zantac, or Axid). Compared to vitamin supplement non-users, supplemented residents had significantly higher mean B12 (p < 0.0001) and erythrocyte folate (p < 0.05) concentrations and significantly lower mean homocysteine (p < 0.01) levels; 229.1 versus 423.6 pmol/L for B12, 882.9 versus 1043.6 nmol/L for erythrocyte folate and 14.4 versus 12.0 micromol/L for homocysteine. CONCLUSION: Given the prevalence data on vitamin status in this sample population, the possible benefits of vitamin supplementation should be considered in clinical intervention studies using these populations of elderly. PMID- 16351717 TI - Gene capture prediction and overlap estimation in EST sequencing from one or multiple libraries. AB - BACKGROUND: In expressed sequence tag (EST) sequencing, we are often interested in how many genes we can capture in an EST sample of a targeted size. This information provides insights to sequencing efficiency in experimental design, as well as clues to the diversity of expressed genes in the tissue from which the library was constructed. RESULTS: We propose a compound Poisson process model that can accurately predict the gene capture in a future EST sample based on an initial EST sample. It also allows estimation of the number of expressed genes in one cDNA library or co-expressed in two cDNA libraries. The superior performance of the new prediction method over an existing approach is established by a simulation study. Our analysis of four Arabidopsis thaliana EST sets suggests that the number of expressed genes present in four different cDNA libraries of Arabidopsis thaliana varies from 9155 (root) to 12005 (silique). An observed fraction of co-expressed genes in two different EST sets as low as 25% can correspond to an actual overlap fraction greater than 65%. CONCLUSION: The proposed method provides a convenient tool for gene capture prediction and cDNA library property diagnosis in EST sequencing. PMID- 16351718 TI - Unhappiness and dissatisfaction in doctors cannot be predicted by selectors from medical school application forms: a prospective, longitudinal study. AB - BACKGROUND: Personal statements and referees' reports are widely used on medical school application forms, particularly in the UK, to assess the suitability of candidates for a career in medicine. However there are few studies which assess the validity of such information for predicting unhappiness or dissatisfaction with a career in medicine. Here we combine data from a long-term prospective study of medical student selection and training, with an experimental approach in which a large number of assessors used a paired comparison technique to predict outcome. METHODS: Data from a large-scale prospective study of students applying to UK medical schools in 1990 were used to identify 40 pairs of doctors, matched by sex, for whom personal statements and referees' reports were available, and who in a 2002/3 follow-up study, one pair member was very satisfied and the other very dissatisfied with medicine as a career. In 2005, 96 assessors, who were experienced medical school selectors, doctors, medical students or psychology students, used information from the doctors' original applications to judge which member of each pair of doctors was the happier, more satisfied doctor. RESULTS: None of the groups of assessors were significantly different from chance expectations in using applicants' personal statements and the referees' reports to predict actual future satisfaction or dissatisfaction, the distribution being similar to binomial expectations. However judgements of pairs of application forms from pairs of doctors showed a non-binomial distribution, indicating consensus among assessors as to which doctor would be the happy doctor (although the consensus was wrong in half the cases). Assessors taking longer to do the task concurred more. Consensus judgements seem mainly to be based on referees' predictions of academic achievement (even though academic achievement is not actually a valid predictor of happiness or satisfaction). CONCLUSION: Although widely used in medical student selection to assess motivation, interest and commitment to a medical career, the personal statement and the referee's report cannot validly be used by assessors, including experienced medical school selectors, to identify doctors who will subsequently be dissatisfied with a medical career. PMID- 16351719 TI - Reproducibility of cervical range of motion in patients with neck pain. AB - BACKGROUND: Reproducibility measurements of the range of motion are an important prerequisite for the interpretation of study results. The aim of the study is to assess the intra-rater and inter-rater reproducibility of the measurement of active Range of Motion (ROM) in patients with neck pain using the Cybex Electronic Digital Inclinometer-320 (EDI-320). METHODS: In an outpatient clinic in a primary care setting 32 patients with at least 2 weeks of pain and/or stiffness in the neck were randomly assessed, in a test- retest design with blinded raters using a standardized measurement protocol. Cervical flexion extension, lateral flexion and rotation were assessed. RESULTS: Reliability expressed by the Intraclass Correlation Coefficient (ICC) was 0.93 (lateral flexion) or higher for intra-rater reliability and 0.89 (lateral flexion) or higher for inter-rater reliability. The 95% limits of agreement for intra-rater agreement, expressing the range of the differences between two ratings were -2.5 +/- 11.1 degrees for flexion-extension, -0.1 +/- 10.4 degrees for lateral flexion and -5.9 +/- 13.5 degrees for rotation. For inter-rater agreement the limits of agreement were 3.3 +/- 17.0 degrees for flexion-extension, 0.5 +/- 17.0 degrees for lateral flexion and -1.3 +/- 24.6 degrees for rotation. CONCLUSION: In general, the intra-rater reproducibility and the inter-rater reproducibility were good. We recommend to compare the reproducibility and clinical applicability of the EDI-320 inclinometer with other cervical ROM measures in symptomatic patients. PMID- 16351721 TI - Psychiatric patients turnaround times in the emergency department. AB - BACKGROUND: To analyze the turnaround times of psychiatric patients within the Emergency Department (ED) from registration to discharge or hospitalization in a University Hospital in 2002. METHODS: Data from a one-year period of psychiatric admissions to the emergency service at a University Hospital were monitored and analyzed focused on turnaround times within the ED. Information on patients variables such as age, sex, diagnosis, consultations and diagnostic procedures were extracted from the patients' charts. RESULTS: From 34.058 patients seen in the ED in 2002, 2632 patients were examined by psychiatrists on duty. Mean turnaround time in the ED was 123 (SD 97) minutes (median 95). Patients to be hospitalized on a psychiatric ward stayed shorter within the ED, patients who later were admitted to another faculty, were treated longer in the ED. Patients with cognitive or substance related disorders stayed longer in the ED than patients with other psychiatric diagnoses. The number of diagnostic procedures and consultations increased the treatment time significantly. CONCLUSION: As the number of patients within the examined ED increases every year, the relevant variables responsible for longer or complicated treatments were assessed in order to appropriately change routine procedures without loss of medical standards. Using this basic data, comparisons with the following years and other hospitals will help to define where the benchmark of turnaround times for psychiatric emergency services might be. PMID- 16351722 TI - Nutrient estimation from an FFQ developed for a Black Zimbabwean population. AB - BACKGROUND: There is little information in the literature on methods of food composition database development to calculate nutrient intake from food frequency questionnaire (FFQ) data. The aim of this study is to describe the development of an FFQ and a food composition table to calculate nutrient intake in a Black Zimbabwean population. METHODS: Trained interviewers collected 24-hour dietary recalls (24 hr DR) from high and low income families in urban and rural Zimbabwe. Based on these data and input from local experts we developed an FFQ, containing a list of frequently consumed foods, standard portion sizes, and categories of consumption frequency. We created a food composition table of the foods found in the FFQ so that we could compute nutrient intake. We used the USDA nutrient database as the main resource because it is relatively complete, updated, and easily accessible. To choose the food item in the USDA nutrient database that most closely matched the nutrient content of the local food we referred to a local food composition table. RESULTS: Almost all the participants ate sadza (maize porridge) at least 5 times a week, and about half had matemba (fish) and caterpillar more than once a month. Nutrient estimates obtained from the FFQ data by using the USDA and Zimbabwean food composition tables were similar for total energy intake intra class correlation (ICC) = 0.99, and carbohydrate (ICC = 0.99), but different for vitamin A (ICC = 0.53), and total folate (ICC = 0.68). CONCLUSION: We have described a standardized process of FFQ and food composition database development for a Black Zimbabwean population. PMID- 16351720 TI - Increased synaptic microtubules and altered synapse development in Drosophila sec8 mutants. AB - BACKGROUND: Sec8 is highly expressed in mammalian nervous systems and has been proposed to play a role in several aspects of neural development and function, including neurite outgrowth, calcium-dependent neurotransmitter secretion, trafficking of ionotropic glutamate receptors and regulation of neuronal microtubule assembly. However, these models have never been tested in vivo. Nervous system development and function have not been described after mutation of sec8 in any organism. RESULTS: We identified lethal sec8 mutants in an unbiased forward genetic screen for mutations causing defects in development of glutamatergic Drosophila neuromuscular junctions (NMJs). The Drosophila NMJ is genetically malleable and accessible throughout development to electrophysiology and immunocytochemistry, making it ideal for examination of the sec8 mutant synaptic phenotype. We developed antibodies to Drosophila Sec8 and showed that Sec8 is abundant at the NMJ. In our sec8 null mutants, in which the sec8 gene is specifically deleted, Sec8 immunoreactivity at the NMJ is eliminated but immunoblots reveal substantial maternal contribution in the rest of the animal. Contrary to the hypothesis that Sec8 is required for neurite outgrowth or synaptic terminal growth, immunocytochemical examination revealed that sec8 mutant NMJs developed more branches and presynaptic terminals during larval development, compared to controls. Synaptic electrophysiology showed no evidence that Sec8 is required for basal neurotransmission, though glutamate receptor trafficking was mildly disrupted in sec8 mutants. The most dramatic NMJ phenotype in sec8 mutants was an increase in synaptic microtubule density, which was approximately doubled compared to controls. CONCLUSION: Sec8 is abundant in the Drosophila NMJ. Sec8 is required in vivo for regulation of synaptic microtubule formation, and (probably secondarily) regulation of synaptic growth and glutamate receptor trafficking. We did not find any evidence that Sec8 is required for basal neurotransmission. PMID- 16351723 TI - AIDS, conflict and the media in Africa: risks in reporting bad data badly. AB - BACKGROUND: Conflict, poverty and HIV disproportionately affect people in sub Saharan Africa. The manner in which governments, national and international organisations and the media report on the HIV epidemic in situations of conflict, post-conflict and reconstruction can have unintended and negative consequences for those affected populations. The media in particular has a huge influence on how the world observes and reacts to the HIV epidemic among conflict-affected and displaced populations. DISCUSSION: Three case studies focused on Sudan, Uganda and Guinea describe what the media reported and why the reports were incomplete, misleading or incorrect. The exploration of possible ways to ensure that the media do not unwittingly inflame delicate and complicated situations of HIV among conflict-affected and displaced populations is then undertaken using epidemiological and journalistic principles. The discussion is divided into four sections: 1) Avoid stigmatising statements and ensure a balanced view; 2) Avoid accurate but misleading statements; 3) Avoid inaccurate statements by clearly stating sources and verifying their credibility; and 4) Do not repeat data and conclusions from other news sources without checking their accuracy. The aim of this manuscript is to stimulate awareness and debate among persons and organisations working on HIV/AIDS as well as the media in order to improve dialogue and ultimately to reduce stigma and discrimination amongst an already vulnerable group--conflict-affected and displaced persons. SUMMARY: The media and humanitarian organisations have published misleading and inaccurate HIV data and statements on conflict-affected and displaced populations in Sudan, Uganda and Guinea. Given the unique characteristics of the HIV epidemic and conflict affected and displaced populations, the media have a special obligation to report in a balanced and non-discriminatory manner that may go beyond the accepted standards of journalism. The media may wish to have the HIV data and their interpretation reviewed by technical experts before going to press. Specific training for reporters and editors regarding ethical issues and basic epidemiological methods may help them to better understand the complexity of the situation and report more accurately; similar training for media watch groups and human rights organisations may improve the monitoring of such situations and possibly reduce misreporting and subsequent discrimination. More rigorous HIV biological and behavioural surveillance should be undertaken in situations of conflict and displacement and humanitarian guidelines should be integrated with guidance on media relations and reporting responsibilities of humanitarian agencies. Finally, humanitarian agencies must ensure the data they release are sound and that any biases are clearly stated. Improved communication with the media will help to ensure more accurate reporting and interpretation. PMID- 16351725 TI - Simple technique for evacuation of traumatic subcutaneous haematomas under tension. AB - BACKGROUND: Traumatic subcutaneous haematomas are common cases in the accident and emergency department. Occasionally, urgent evacuation is required to prevent skin necrosis. METHODS: We present a simple and safe technique, based on a principle similar to liposuction to evacuate traumatic subcutaneous haematomas under tension. Instruments readily available in the accident and emergency department are used without the need of general anaesthesia. RESULTS: The technique was performed in six cases without complication such as infection or re collection of the haematoma under tension. We present two typical scenarios where urgent evacuation was indicated and the technique performed. CONCLUSION: The technique is useful as an urgent and safe procedure in the accident and emergency department setting. PMID- 16351724 TI - SP-A binds alpha1-antitrypsin in vitro and reduces the association rate constant for neutrophil elastase. AB - BACKGROUND: Alpha1-antitrypsin and surfactant protein-A (SP-A) are major lung defense proteins. With the hypothesis that SP-A could bind alpha1-antitrypsin, we designed a series of in vitro experiments aimed at investigating the nature and consequences of such an interaction. METHODS AND RESULTS: At an alpha1 antitrypsin:SP-A molar ratio of 1:1, the interaction resulted in a calcium dependent decrease of 84.6% in the association rate constant of alpha1 antitrypsin for neutrophil elastase. The findings were similar when SP-A was coupled with the Z variant of alpha1-antitrypsin. The carbohydrate recognition domain of SP-A appeared to be a major determinant of the interaction, by recognizing alpha1-antitrypsin carbohydrate chains. However, binding of SP-A carbohydrate chains to the alpha1-antitrypsin amino acid backbone and interaction between carbohydrates of both proteins are also possible. Gel filtration chromatography and turnover per inactivation experiments indicated that one part of SP-A binds several molar parts of alpha1-antitrypsin. CONCLUSION: We conclude that the binding of SP-A to alpha1-antitrypsin results in a decrease of the inhibition of neutrophil elastase. This interaction could have potential implications in the physiologic regulation of alpha1-antitrypsin activity, in the pathogenesis of pulmonary emphysema, and in the defense against infectious agents. PMID- 16351726 TI - Adipose energy stores, physical work, and the metabolic syndrome: lessons from hummingbirds. AB - Hummingbirds and other nectar-feeding, migratory birds possess unusual adaptive traits that offer important lessons concerning obesity, diabetes and the metabolic syndrome. Hummingbirds consume a high sugar diet and have fasting glucose levels that would be severely hyperglycemic in humans, yet these nectar fed birds recover most glucose that is filtered into the urine. Hummingbirds accumulate over 40% body fat shortly before migrations in the spring and autumn. Despite hyperglycemia and seasonally elevated body fat, the birds are not known to become diabetic in the sense of developing polyuria (glucosuria), polydipsia and polyphagia. The tiny (3-4 g) Ruby-throated hummingbird has among the highest mass-specific metabolic rates known, and loses most of its stored fat in 20 h by flying up to 600 miles across the Gulf of Mexico. During the breeding season, it becomes lean and maintains an extremely accurate energy balance. In addition, hummingbirds can quickly enter torpor and reduce resting metabolic rates by 10 fold. Thus, hummingbirds are wonderful examples of the adaptive nature of fat tissue, and may offer lessons concerning prevention of metabolic syndrome in humans. PMID- 16351727 TI - A method for accurate detection of genomic microdeletions using real-time quantitative PCR. AB - BACKGROUND: Quantitative Polymerase Chain Reaction (qPCR) is a well-established method for quantifying levels of gene expression, but has not been routinely applied to the detection of constitutional copy number alterations of human genomic DNA. Microdeletions or microduplications of the human genome are associated with a variety of genetic disorders. Although, clinical laboratories routinely use fluorescence in situ hybridization (FISH) to identify such cryptic genomic alterations, there remains a significant number of individuals in which constitutional genomic imbalance is suspected, based on clinical parameters, but cannot be readily detected using current cytogenetic techniques. RESULTS: In this study, a novel application for real-time qPCR is presented that can be used to reproducibly detect chromosomal microdeletions and microduplications. This approach was applied to DNA from a series of patient samples and controls to validate genomic copy number alteration at cytoband 22q11. The study group comprised 12 patients with clinical symptoms of chromosome 22q11 deletion syndrome (22q11DS), 1 patient trisomic for 22q11 and 4 normal controls. 6 of the patients (group 1) had known hemizygous deletions, as detected by standard diagnostic FISH, whilst the remaining 6 patients (group 2) were classified as 22q11DS negative using the clinical FISH assay. Screening of the patients and controls with a set of 10 real time qPCR primers, spanning the 22q11.2-deleted region and flanking sequence, confirmed the FISH assay results for all patients with 100% concordance. Moreover, this qPCR enabled a refinement of the region of deletion at 22q11. Analysis of DNA from chromosome 22 trisomic sample demonstrated genomic duplication within 22q11. CONCLUSION: In this paper we present a qPCR approach for the detection of chromosomal microdeletions and microduplications. The strategic use of in silico modelling for qPCR primer design to avoid regions of repetitive DNA, whilst providing a level of genomic resolution greater than standard cytogenetic assays. The implementation of qPCR detection in clinical laboratories will address the need to replace complex, expensive and time consuming FISH screening to detect genomic microdeletions or duplications of clinical importance. PMID- 16351728 TI - Socio economic crisis and mortality. Epidemiological testimony of the financial collapse of Argentina. AB - BACKGROUND: Natural disasters, war, and terrorist attacks, have been linked to cardiac mortality. We sought to investigate whether a major financial crisis may impact on the medical management and outcomes of acute coronary syndromes. METHODS: We analyzed the Argentine cohort of the international multicenter Global Registry of Acute Coronary Events (GRACE). The primary objective was to estimate if there was an association between the financial crisis period (April 1999 to December 2002) and in- hospital cardiovascular mortality, with the post-crisis period (January 2003 to September 2004) as the referent. Each period was defined according to the evolution of the Gross Domestic Product. We investigated the demographic characteristics, diagnostic and therapeutic procedures, morbidity and mortality. RESULTS: We analyzed data from 3220 patients, 2246 (69.8%) patients in the crisis period and 974 (30.2%) in the post-crisis frame. The distribution of demographic and clinical baseline characteristics were not significantly different between both periods. During the crisis period the incidence of in hospital myocardial infarction was higher (6.9% Vs 2.9%; p value < 0.0001), as well as congestive heart failure (16% Vs 11%; p value < 0.0001). Time to intervention with angioplasty was longer during the crisis, especially among public sites (median 190 min Vs 27 min). The incidence proportion of mortality during hospitalization was 6.2% Vs 5.1% after crisis. The crude OR for mortality was 1.2 (95% C.I. 0.87, 1.7). The odds for mortality were higher among private institutions {1.9 (95% C.I. 0.9, 3.8)} than for public centers {1.2 (95% C.I. 0.83, 1.79)}. We did not observe a significant interaction between type of hospital and crisis. CONCLUSION: Our findings suggest that the financial crisis may have had a negative impact on cardiovascular mortality during hospitalization, and higher incidence of medical complications. PMID- 16351729 TI - Cortex reorganization of Xenopus laevis eggs in strong static magnetic fields. AB - Observations of magnetic field effects on biological systems have often been contradictory. For amphibian eggs, a review of the available literature suggests that part of the discrepancies might be resolved by considering a previously neglected parameter for morphological alterations induced by magnetic fields--the jelly layers that normally surround the egg and are often removed in laboratory studies for easier cell handling. To experimentally test this hypothesis, we observed the morphology of fertilizable Xenopus laevis eggs with and without jelly coat that were subjected to static magnetic fields of up to 9.4 T for different periods of time. A complex reorganization of cortical pigmentation was found in dejellied eggs as a function of the magnetic field and the field exposure time. Initial pigment rearrangements could be observed at about 0.5 T, and less than 3 T are required for the effects to fully develop within two hours. No effect was observed when the jelly layers of the eggs were left intact. These results suggest that the action of magnetic fields might involve cortical pigments or associated cytoskeletal structures normally held in place by the jelly layers and that the presence of the jelly layer should indeed be included in further studies of magnetic field effects in this system. PMID- 16351730 TI - Combining short stent implantation and drug-eluting stenting for routine use yields a low restenosis rate. AB - BACKGROUND: Stent length serves as a predictor of restenosis in use of bare metal stents (BMS). This has been demonstrated in a feasibility study that used a single short BMS implant (<9 mm) in a high proportion of lesions; the study observed a low rate of restenosis. METHODS: We performed a pilot prospective study to investigate in a series of consecutive patients the immediate and long term effects of implantation of either 1) a single short BMS for all lesions with low probability of restenosis or 2) a drug-eluting stent (DES) for all other lesions. RESULTS: The 200 patients studied had 236 coronary artery lesions that were treated with short BMS in 168/236 patients (71.2%) and with DES in 68/236 patients (28.8%). Angiographic success was achieved in 230/236 lesions (97.5%) and procedural success in 194/200 patients (97.0%). Restenosis occurred in 15/153 lesions (9.8%) after short BMS, in 3/62 lesions (4.8%) after DES, and in 18/215 of all lesions (8.4%) angiographically controlled after six to eight months. Target vessel revascularization was performed in 16/218 lesion (7.4%). CONCLUSION: Most of the coronary artery lesions in this small group of consecutive patients were treated sufficiently with a single BMS implant. This differential approach of treating suitable lesions in medium- to large-sized vessels with a single short BMS device and treating all other lesions with a DES implant resulted in a low incidence of restenosis. PMID- 16351731 TI - Metallothionien 3 expression is frequently down-regulated in oesophageal squamous cell carcinoma by DNA methylation. AB - BACKGROUND: Metallothionein 3 (MT3) inhibits growth in a variety of cell types. We measured MT3 gene expression by RT-PCR, and DNA methylation in the MT3 promoter by combined bisulphite restriction analysis, in four oesophageal cancer cell lines and the resected oesophagus from 64 patients with oesophageal squamous cell carcinoma (SCC). RESULTS: MT3 expression was not detected in one of the four oesophageal cell lines. The MT3 promoter was methylated in all of the oesophageal cell lines, but the degree of methylation was greater in the non-expressing cell line. After treatment with 5-aza-2'-deoxycytidine there was a reduction in the degree of methylation, and an increase in MT3 expression, in each of the cell lines (p < 0.01). Methylation was detected in 52% (33 of 64) of primary SCC and 3% (2 of 62) of histologically normal resection margins. MT3 expression was measured in 29 tumours, 17 of which had methylation of MT3. The expression of MT3 was significantly less in the methylated tumours compared to either the unmethylated tumours (p = 0.03), or the matched margin (p = 0.0005). There was not a significant difference in MT3 expression between the tumour and the margin from patients with unmethylated tumour. No correlations were observed between methylation of MT3 and survival time, patient age, gender, smoking or drinking history, tumour stage, volume, or lymph node involvement. CONCLUSION: We conclude that MT3 expression is frequently down-regulated in oesophageal SCC, by DNA methylation, but that this is not a prognostic indicator. PMID- 16351732 TI - A rare case of disseminated cutaneous zoster in an immunocompetent patient. AB - BACKGROUND: Disseminated cutaneous herpes zoster in healthy persons is uncommon, though it has been described in immunocompromised patients. CASE PRESENTATION: We describe a case of disseminated cutaneous herpes zoster in an elderly man with no apparent immunosuppressive condition. The patient was treated successfully with intravenous Acyclovir. CONCLUSION: We suggest that disseminated zoster can occur in an immunocompetent host and should be promptly recognized and treated to prevent serious complications. PMID- 16351733 TI - The Hospital Anxiety and Depression Rating Scale: a cross-sectional study of psychometrics and case finding abilities in general practice. AB - BACKGROUND: General practitioners' (GPs) diagnostic skills lead to underidentification of generalized anxiety disorders (GAD) and major depressive episodes (MDE). Supplement of brief questionnaires could improve the diagnostic accuracy of GPs for these common mental disorders. The aims of this study were to examine the usefulness of The Hospital Anxiety and Depression Rating Scale (HADS) for GPs by: 1) Examining its psychometrics in the GPs' setting; 2) Testing its case-finding properties compared to patient-rated GAD and MDE (DSM-IV); and 3) Comparing its case finding abilities to that of the GPs using Clinical Global Impression-Severity (CGI-S) rating. METHODS: In a cross-sectional survey study 1,781 patients in three consecutive days in September 2001 attended 141 GPs geographically spread in Norway. Sensitivity, specificity, optimal cut off score, and Area under the curve (AUC) for the HADS and the CGI-S were calculated with Generalized Anxiety Questionnaire (GAS-Q) as reference standard for GAD, and Depression Screening Questionnaire (DSQ) for MDE. RESULTS: The HADS-A had optimal cut off > or =8 (sensitivity 0.89, specificity 0.75), AUC 0.88 and 76% of patients were correctly classified in relation to GAD. The HADS-D had by optimal cut off > or =8 (sensitivity 0.80 and specificity 0.88) AUC 0.93 and 87% of the patients were correctly classified in relation to MDE. Proportions of the total correctly classified at the CGI-S optimal cut-off > or =3 were 83% of patients for GAD and 81% for MDE. CONCLUSION: The results indicate that addition of the patients' HADS scores to GPs' information could improve their diagnostic accuracy of GAD and MDE. PMID- 16351734 TI - Looking for pyromania: characteristics of a consecutive sample of Finnish male criminals with histories of recidivist fire-setting between 1973 and 1993. AB - BACKGROUND: As pyromania is a rare diagnosis with questionable validity, we aimed to describe a forensic psychiatric population of arson recidivists. METHODS: The medical records as well as the forensic psychiatric examination statements of 90 arson recidivists referred for pretrial psychiatric assessment in Helsinki University Hospital Department of Forensic Psychiatry between 1973 and 1993 were reviewed. RESULTS: The most important diagnostic categories of arson recidivists were personality disorders, psychosis and mental retardation, often with comorbid alcoholism. In all, 68% of arsonists were under alcohol intoxication during the index crime. Psychotic as well as mentally retarded persons with repeated fire setting behaviour were mostly "pure arsonists"--persons guilty only of arsons during their criminal careers. Arson recidivists with personality disorder, in contrast, often exhibited various types of criminal behaviour and arson appeared to be only one expression of a wide range of criminal activity. Comorbid alcoholism was apparently a more rarely observed phenomenon among pure arsonists than in "nonpure arsonists". We found only three subjects fulfilling the present diagnostic criteria for pyromania. CONCLUSION: Using the criteria of the DSM-IV TR, pyromania must be regarded as an extremely rare phenomenon. Especially the question of substance intoxication as an exclusion criterion for pyromania should be reconsidered. PMID- 16351736 TI - Modelling the interaction of steroid receptors with endocrine disrupting chemicals. AB - BACKGROUND: The organic polychlorinated compounds like dichlorodiphenyltrichloroethane with its metabolites and polychlorinated biphenyls are a class of highly persistent environmental contaminants. They have been recognized to have detrimental health effects both on wildlife and humans acting as endocrine disrupters due to their ability of mimicking the action of the steroid hormones, and thus interfering with hormone response. There are several experimental evidences that they bind and activate human steroid receptors. However, despite the growing concern about the toxicological activity of endocrine disrupters, molecular data of the interaction of these compounds with biological targets are still lacking. RESULTS: We have used a flexible docking approach to characterize the molecular interaction of seven endocrine disrupting chemicals with estrogen, progesterone and androgen receptors in the ligand-binding domain. All ligands docked in the buried hydrophobic cavity corresponding to the hormone steroid pocket. The interaction was characterized by multiple hydrophobic contacts involving a different number of residues facing the binding pocket, depending on ligands orientation. The EDC ligands did not display a unique binding mode, probably due to their lipophilicity and flexibility, which conferred them a great adaptability into the hydrophobic and large binding pocket of steroid receptors. CONCLUSION: Our results are in agreement with toxicological data on binding and allow to describe a pattern of interactions for a group of ECD to steroid receptors suggesting the requirement of a hydrophobic cavity to accommodate these chlorine carrying compounds. Although the affinity is lower than for hormones, their action can be brought about by a possible synergistic effect. PMID- 16351737 TI - A quantization method based on threshold optimization for microarray short time series. AB - BACKGROUND: Reconstructing regulatory networks from gene expression profiles is a challenging problem of functional genomics. In microarray studies the number of samples is often very limited compared to the number of genes, thus the use of discrete data may help reducing the probability of finding random associations between genes. RESULTS: A quantization method, based on a model of the experimental error and on a significance level able to compromise between false positive and false negative classifications, is presented, which can be used as a preliminary step in discrete reverse engineering methods. The method is tested on continuous synthetic data with two discrete reverse engineering methods: Reveal and Dynamic Bayesian Networks. CONCLUSION: The quantization method, evaluated in comparison with two standard methods, 5% threshold based on experimental error and rank sorting, improves the ability of Reveal and Dynamic Bayesian Networks to identify relations among genes. PMID- 16351738 TI - A new decoding algorithm for hidden Markov models improves the prediction of the topology of all-beta membrane proteins. AB - BACKGROUND: Structure prediction of membrane proteins is still a challenging computational problem. Hidden Markov models (HMM) have been successfully applied to the problem of predicting membrane protein topology. In a predictive task, the HMM is endowed with a decoding algorithm in order to assign the most probable state path, and in turn the labels, to an unknown sequence. The Viterbi and the posterior decoding algorithms are the most common. The former is very efficient when one path dominates, while the latter, even though does not guarantee to preserve the HMM grammar, is more effective when several concurring paths have similar probabilities. A third good alternative is 1-best, which was shown to perform equal or better than Viterbi. RESULTS: In this paper we introduce the posterior-Viterbi (PV) a new decoding which combines the posterior and Viterbi algorithms. PV is a two step process: first the posterior probability of each state is computed and then the best posterior allowed path through the model is evaluated by a Viterbi algorithm. CONCLUSION: We show that PV decoding performs better than other algorithms when tested on the problem of the prediction of the topology of beta-barrel membrane proteins. PMID- 16351739 TI - A neural strategy for the inference of SH3 domain-peptide interaction specificity. AB - BACKGROUND: The SH3 domain family is one of the most representative and widely studied cases of so-called Peptide Recognition Modules (PRM). The polyproline II motif PxxP that generally characterizes its ligands does not reflect the complex interaction spectrum of the over 1500 different SH3 domains, and the requirement of a more refined knowledge of their specificity implies the setting up of appropriate experimental and theoretical strategies. Due to the limitations of the current technology for peptide synthesis, several experimental high throughput approaches have been devised to elucidate protein-protein interaction mechanisms. Such approaches can rely on and take advantage of computational techniques, such as regular expressions or position specific scoring matrices (PSSMs) to pre-process entire proteomes in the search for putative SH3 targets. In this regard, a reliable inference methodology to be used for reducing the sequence space of putative binding peptides represents a valuable support for molecular and cellular biologists. RESULTS: Using as benchmark the peptide sequences obtained from in vitro binding experiments, we set up a neural network model that performs better than PSSM in the detection of SH3 domain interactors. In particular our model is more precise in its predictions, even if its performance can vary among different SH3 domains and is strongly dependent on the number of binding peptides in the benchmark. CONCLUSION: We show that a neural network can be more effective than standard methods in SH3 domain specificity detection. Neural classifiers identify general SH3 domain binders and domain specific interactors from a PxxP peptide population, provided that there are a sufficient proportion of true positives in the training sets. This capability can also improve peptide selection for library definition in array experiments. Further advances can be achieved, including properly encoded domain sequences and structural information as input for a global neural network. PMID- 16351740 TI - Mining published lists of cancer related microarray experiments: identification of a gene expression signature having a critical role in cell-cycle control. AB - BACKGROUND: Routine application of gene expression microarray technology is rapidly producing large amounts of data that necessitate new approaches of analysis. The analysis of a specific microarray experiment profits enormously from cross-comparing to other experiments. This process is generally performed by numerical meta-analysis of published data where the researcher chooses the datasets to be analyzed based on assumptions about the biological relations of published datasets to his own data, thus severely limiting the possibility of finding surprising connections. Here we propose using a repository of published gene lists for the identification of interesting datasets to be subjected to more detailed numerical analysis. RESULTS: We have compiled lists of genes that have been reported as differentially regulated in cancer related microarray studies. We searched these gene lists for statistically significant overlaps with lists of genes regulated by the tumor suppressors p16 and pRB. We identified a highly significant overlap of p16 and pRB target genes with genes regulated by the EWS/FLI fusion protein. Detailed numerical analysis of these data identified two sets of genes with clearly distinct roles in the G1/S and the G2/M phases of the cell cycle, as measured by enrichment of Gene Ontology categories. CONCLUSION: We show that mining of published gene lists in the absence of numerical detail about gene expression levels constitutes a fast, easy to perform, widely applicable, and unbiased route towards the identification of biologically related gene expression microarray datasets. PMID- 16351742 TI - ESTree db: a tool for peach functional genomics. AB - BACKGROUND: The ESTree db http://www.itb.cnr.it/estree/ represents a collection of Prunus persica expressed sequenced tags (ESTs) and is intended as a resource for peach functional genomics. A total of 6,155 successful EST sequences were obtained from four in-house prepared cDNA libraries from Prunus persica mesocarps at different developmental stages. Another 12,475 peach EST sequences were downloaded from public databases and added to the ESTree db. An automated pipeline was prepared to process EST sequences using public software integrated by in-house developed Perl scripts and data were collected in a MySQL database. A php-based web interface was developed to query the database. RESULTS: The ESTree db version as of April 2005 encompasses 18,630 sequences representing eight libraries. Contig assembly was performed with CAP3. Putative single nucleotide polymorphism (SNP) detection was performed with the AutoSNP program and a search engine was implemented to retrieve results. All the sequences and all the contig consensus sequences were annotated both with blastx against the GenBank nr db and with GOblet against the viridiplantae section of the Gene Ontology db. Links to NiceZyme (Expasy) and to the KEGG metabolic pathways were provided. A local BLAST utility is available. A text search utility allows querying and browsing the database. Statistics were provided on Gene Ontology occurrences to assign sequences to Gene Ontology categories. CONCLUSION: The resulting database is a comprehensive resource of data and links related to peach EST sequences. The Sequence Report and Contig Report pages work as the web interface core structures, giving quick access to data related to each sequence/contig. PMID- 16351741 TI - In silico functional characterization of a double histone fold domain from the Heliothis zea virus 1. AB - BACKGROUND: Histones are short proteins involved in chromatin packaging; in eukaryotes, two H2a-H2b and H3-H4 histone dimers form the nucleosomal core, which acts as the fundamental DNA-packaging element. The double histone fold is a rare globular protein fold in which two consecutive regions characterized by the typical structure of histones assemble together, thus originating a histone pseudodimer. This fold is included in a few prokaryotic histones and in the regulatory region of guanine nucleotide exchange factors of the Sos family. For the prokaryotic histones, there is no direct structural counterpart in the nucleosomal core particle, while the pseudodimer from Sos proteins is very similar to the dimer formed by histones H2a and H2b. RESULTS: The absence of a H3 H4-like histone pseudodimer in the available structural databases prompted us to search for proteins that could assume such fold. The application of several secondary structure prediction and fold recognition methods allowed to show that the viral protein gi|22788712 is compatible with the structure of a H3-H4-like histone pseudodimer. Further in silico analyses revealed that this protein module could retain the ability of mediating protein-DNA interactions, and could consequently act as a DNA-binding domain. CONCLUSION: Our results suggest a possible functional role in viral pathogenicity for this novel double histone fold domain; thus, the computational analyses here reported will be helpful in directing future biochemical studies on gi|22788712 protein. PMID- 16351743 TI - Browsing isolated population data. AB - BACKGROUND: In our studies of genetically isolated populations in a remote mountain area in the center of Sardinia (Italy), we found that 80-85% of the inhabitants of each village belong to a single huge pedigree with families strictly connected to each other through hundreds of loops. Moreover, intermarriages between villages join pedigrees of different villages through links that make family trees even more complicated. Unfortunately, none of the commonly used pedigree drawing tools are able to draw the complete pedigree, whereas it is commonly accepted that the visual representation of families is very important as it helps researchers in identifying clusters of inherited traits and genotypes. We had a representation issue that compels researchers to work with subsets extracted from the overall genealogy, causing a serious loss of information on familiar relationships. To visually explore such complex pedigrees, we developed PedNavigator, a browser for genealogical databases properly suited for genetic studies. RESULTS: The PedNavigator is useful for genealogical research due to its capacity to represent family relations between persons and to make a visual verification of the links during family history reconstruction. As for genetic studies, it is helpful to follow propagation of a specific set of genetic markers (haplotype), or to select people for linkage analysis, showing relations between various branch of a family tree of affected subjects. AVAILABILITY: PedNavigator is an application integrated into a Framework designed to handle data for human genetic studies based on the Oracle platform. To allow the use of PedNavigator also to people not owning the same required informatics infrastructure or systems, we developed PedNavigator Lite with mainly the same features of the integrated one, based on MySQL database server. This version is free for academic users, and it is available for download from our site http://www.shardna.com. PMID- 16351744 TI - Inherited disorder phenotypes: controlled annotation and statistical analysis for knowledge mining from gene lists. AB - BACKGROUND: Analysis of inherited diseases and their associated phenotypes is of great importance to gain knowledge of underlying genetic interactions and could ultimately give clinically useful insights into disease processes, including complex diseases influenced by multiple genetic loci. Nevertheless, to date few computational contributions have been proposed for this purpose, mainly due to lack of controlled clinical information easily accessible and structured for computational genome-wise analyses. To allow performing phenotype analyses of inherited disorder related genes we implemented new original modules within GFINDer http://www.bioinformatics.polimi.it/GFINDer/, a Web system we previously developed that dynamically aggregates functional annotations of user uploaded gene lists and allows performing their statistical analysis and mining. RESULTS: New GFINDer modules allow annotating large numbers of user classified biomolecular sequence identifiers with morbidity and clinical information, classifying them according to genetic disease phenotypes and their locations of occurrence, and statistically analyzing the obtained classifications. To achieve this we exploited, normalized and structured the information present in textual form in the Clinical Synopsis sections of the Online Mendelian Inheritance in Man (OMIM) databank. Such valuable information delineates numerous signs and symptoms accompanying many genetic diseases and it is divided into phenotype location categories, either by organ system or type of finding. CONCLUSION: Supporting phenotype analyses of inherited diseases and biomolecular functional evaluations, GFINDer facilitates a genomic approach to the understanding of fundamental biological processes and complex cellular mechanisms underlying patho physiological phenotypes. PMID- 16351745 TI - High performance workflow implementation for protein surface characterization using grid technology. AB - BACKGROUND: This study concerns the development of a high performance workflow that, using grid technology, correlates different kinds of Bioinformatics data, starting from the base pairs of the nucleotide sequence to the exposed residues of the protein surface. The implementation of this workflow is based on the Italian Grid.it project infrastructure, that is a network of several computational resources and storage facilities distributed at different grid sites. METHODS: Workflows are very common in Bioinformatics because they allow to process large quantities of data by delegating the management of resources to the information streaming. Grid technology optimizes the computational load during the different workflow steps, dividing the more expensive tasks into a set of small jobs. RESULTS: Grid technology allows efficient database management, a crucial problem for obtaining good results in Bioinformatics applications. The proposed workflow is implemented to integrate huge amounts of data and the results themselves must be stored into a relational database, which results as the added value to the global knowledge. CONCLUSION: A web interface has been developed to make this technology accessible to grid users. Once the workflow has started, by means of the simplified interface, it is possible to follow all the different steps throughout the data processing. Eventually, when the workflow has been terminated, the different features of the protein, like the amino acids exposed on the protein surface, can be compared with the data present in the output database. PMID- 16351746 TI - Regularized Least Squares Cancer classifiers from DNA microarray data. AB - BACKGROUND: The advent of the technology of DNA microarrays constitutes an epochal change in the classification and discovery of different types of cancer because the information provided by DNA microarrays allows an approach to the problem of cancer analysis from a quantitative rather than qualitative point of view. Cancer classification requires well founded mathematical methods which are able to predict the status of new specimens with high significance levels starting from a limited number of data. In this paper we assess the performances of Regularized Least Squares (RLS) classifiers, originally proposed in regularization theory, by comparing them with Support Vector Machines (SVM), the state-of-the-art supervised learning technique for cancer classification by DNA microarray data. The performances of both approaches have been also investigated with respect to the number of selected genes and different gene selection strategies. RESULTS: We show that RLS classifiers have performances comparable to those of SVM classifiers as the Leave-One-Out (LOO) error evaluated on three different data sets shows. The main advantage of RLS machines is that for solving a classification problem they use a linear system of order equal to either the number of features or the number of training examples. Moreover, RLS machines allow to get an exact measure of the LOO error with just one training. CONCLUSION: RLS classifiers are a valuable alternative to SVM classifiers for the problem of cancer classification by gene expression data, due to their simplicity and low computational complexity. Moreover, RLS classifiers show generalization ability comparable to the ones of SVM classifiers also in the case the classification of new specimens involves very few gene expression levels. PMID- 16351747 TI - Systematic analysis of human kinase genes: a large number of genes and alternative splicing events result in functional and structural diversity. AB - BACKGROUND: Protein kinases are a well defined family of proteins, characterized by the presence of a common kinase catalytic domain and playing a significant role in many important cellular processes, such as proliferation, maintenance of cell shape, apoptosis. In many members of the family, additional non-kinase domains contribute further specialization, resulting in subcellular localization, protein binding and regulation of activity, among others. About 500 genes encode members of the kinase family in the human genome, and although many of them represent well known genes, a larger number of genes code for proteins of more recent identification, or for unknown proteins identified as kinase only after computational studies. RESULTS: A systematic in silico study performed on the human genome, led to the identification of 5 genes, on chromosome 1, 11, 13, 15 and 16 respectively, and 1 pseudogene on chromosome X; some of these genes are reported as kinases from NCBI but are absent in other databases, such as KinBase. Comparative analysis of 483 gene regions and subsequent computational analysis, aimed at identifying unannotated exons, indicates that a large number of kinase may code for alternately spliced forms or be incorrectly annotated. An InterProScan automated analysis was performed to study domain distribution and combination in the various families. At the same time, other structural features were also added to the annotation process, including the putative presence of transmembrane alpha helices, and the cystein propensity to participate into a disulfide bridge. CONCLUSION: The predicted human kinome was extended by identifying both additional genes and potential splice variants, resulting in a varied panorama where functionality may be searched at the gene and protein level. Structural analysis of kinase proteins domains as defined in multiple sources together with transmembrane alpha helices and signal peptide prediction provides hints to function assignment. The results of the human kinome analysis are collected in the KinWeb database, available for browsing and searching over the internet, where all results from the comparative analysis and the gene structure annotation are made available, alongside the domain information. Kinases may be searched by domain combinations and the relative genes may be viewed in a graphic browser at various level of magnification up to gene organization on the full chromosome set. PMID- 16351748 TI - HomoMINT: an inferred human network based on orthology mapping of protein interactions discovered in model organisms. AB - BACKGROUND: The application of high throughput approaches to the identification of protein interactions has offered for the first time a glimpse of the global interactome of some model organisms. Until now, however, such genome-wide approaches have not been applied to the human proteome. RESULTS: In order to fill this gap we have assembled an inferred human protein interaction network where interactions discovered in model organisms are mapped onto the corresponding human orthologs. In addition to a stringent assignment to orthology classes based on the InParanoid algorithm, we have implemented a string matching algorithm to filter out orthology assignments of proteins whose global domain organization is not conserved. Finally, we have assessed the accuracy of our own, and related, inferred networks by benchmarking them against i) an assembled experimental interactome, ii) a network derived by mining of the scientific literature and iii) by measuring the enrichment of interacting protein pairs sharing common Gene Ontology annotation. CONCLUSION: The resulting networks are named HomoMINT and HomoMINT_filtered, the latter being based on the orthology table filtered by the domain architecture matching algorithm. They contains 9749 and 5203 interactions respectively and can be analyzed and viewed in the context of the experimentally verified interactions between human proteins stored in the MINT database. HomoMINT is constantly updated to take into account the growing information in the MINT database. PMID- 16351749 TI - NemaFootPrinter: a web based software for the identification of conserved non coding genome sequence regions between C. elegans and C. briggsae. AB - BACKGROUND: NemaFootPrinter (Nematode Transcription Factor Scan Through Philogenetic Footprinting) is a web-based software for interactive identification of conserved, non-exonic DNA segments in the genomes of C. elegans and C. briggsae. It has been implemented according to the following project specifications:a) Automated identification of orthologous gene pairs. b) Interactive selection of the boundaries of the genes to be compared. c) Pairwise sequence comparison with a range of different methods. d) Identification of putative transcription factor binding sites on conserved, non-exonic DNA segments. RESULTS: Starting from a C. elegans or C. briggsae gene name or identifier, the software identifies the putative ortholog (if any), based on information derived from public nematode genome annotation databases. The investigator can then retrieve the genome DNA sequences of the two orthologous genes; visualize graphically the genes' intron/exon structure and the surrounding DNA regions; select, through an interactive graphical user interface, subsequences of the two gene regions. Using a bioinformatics toolbox (Blast2seq, Dotmatcher, Ssearch and connection to the rVista database) the investigator is able at the end of the procedure to identify and analyze significant sequences similarities, detecting the presence of transcription factor binding sites corresponding to the conserved segments. The software automatically masks exons. DISCUSSION: This software is intended as a practical and intuitive tool for the researchers interested in the identification of non-exonic conserved sequence segments between C. elegans and C. briggsae. These sequences may contain regulatory transcriptional elements since they are conserved between two related, but rapidly evolving genomes. This software also highlights the power of genome annotation databases when they are conceived as an open resource and the possibilities offered by seamless integration of different web services via the http protocol. AVAILABILITY: The program is freely available at http://bio.ifom firc.it/NTFootPrinter. PMID- 16351750 TI - Improving interoperability between microbial information and sequence databases. AB - BACKGROUND: Biological resources are essential tools for biomedical research. Their availability is promoted through on-line catalogues. Common Access to Biological Resources and Information (CABRI) is a service for distribution of biological resources and related data collected by 28 European culture collections. Linking this information to bioinformatics databanks can make the collections' holdings more visible after a search in molecular biology databanks and vice-versa. Identification of links to sequence databases can be useful, but annotation and indexing problems, together with compilation errors, immediately arise. In this paper, we present our efforts for the identification of cross references between CABRI catalogues and the EMBL Data Library and related results. RESULTS: An SRS site with both EMBL and CABRI catalogues has been set up. Ad-hoc changes in indexing scripts allowed to achieve homogeneous index keys and SRS link features have been used to identify links between databases. After manual checking and comparison with an alternative procedure, about 67,500 valid cross-references were identified, added to the EMBL Data Library and are now distributed with it. HTML links can be established from EMBL to CABRI network service. Procedures can be executed whenever needed. CONCLUSION: Links between EMBL and CABRI catalogues constitute an improved access to micro-organisms of certified quality and can produce positive effects on biomedical research. Further links between CABRI catalogues and other bioinformatics databases can now easily be defined by using these cross-references. Linking genetic information onto natural resources information may stand model for the integration of other databases containing empirical data on these materials. PMID- 16351751 TI - Web services and workflow management for biological resources. AB - BACKGROUND: The completion of the Human Genome Project has resulted in large quantities of biological data which are proving difficult to manage and integrate effectively. There is a need for a system that is able to automate accesses to remote sites and to "understand" the information that it is managing in order to link data properly. Workflow management systems combined with Web Services are promising Information and Communication Technologies (ICT) tools. Some have already been proposed and are being increasingly applied to the biomedical domain, especially as many biology-related Web Services are now becoming available. Information on biological resources and on genomic sequences mutations are two examples of very specialized datasets that are useful for specific research domains. RESULTS: The architecture of a system that is able to access and execute predefined workflows is presented in this paper. Web Services allowing access to the IARC TP53 Mutation Database and CABRI catalogues of biological resources have been implemented and are available on-line. Example workflows which retrieve data from these Web Services have also been created and are available on-line. CONCLUSION: We present a general architecture and some building blocks for the implementation of a system that is able to remotely execute workflows of biomedical interest and show how this approach can effectively produce useful outputs. The further development and implementation of Web Services allowing access to an exhaustive set of biomedical databases and the creation of effective and useful workflows will improve the automation of in silico analysis. PMID- 16351752 TI - A hybrid genetic-neural system for predicting protein secondary structure. AB - BACKGROUND: Due to the strict relation between protein function and structure, the prediction of protein 3D-structure has become one of the most important tasks in bioinformatics and proteomics. In fact, notwithstanding the increase of experimental data on protein structures available in public databases, the gap between known sequences and known tertiary structures is constantly increasing. The need for automatic methods has brought the development of several prediction and modelling tools, but a general methodology able to solve the problem has not yet been devised, and most methodologies concentrate on the simplified task of predicting secondary structure. RESULTS: In this paper we concentrate on the problem of predicting secondary structures by adopting a technology based on multiple experts. The system performs an overall processing based on two main steps: first, a "sequence-to-structure" prediction is enforced by resorting to a population of hybrid (genetic-neural) experts, and then a "structure-to structure" prediction is performed by resorting to an artificial neural network. Experiments, performed on sequences taken from well-known protein databases, allowed to reach an accuracy of about 76%, which is comparable to those obtained by state-of-the-art predictors. CONCLUSION: The adoption of a hybrid technique, which encompasses genetic and neural technologies, has demonstrated to be a promising approach in the task of protein secondary structure prediction. PMID- 16351753 TI - HmtDB, a human mitochondrial genomic resource based on variability studies supporting population genetics and biomedical research. AB - BACKGROUND: Population genetics studies based on the analysis of mtDNA and mitochondrial disease studies have produced a huge quantity of sequence data and related information. These data are at present worldwide distributed in differently organised databases and web sites not well integrated among them. Moreover it is not generally possible for the user to submit and contemporarily analyse its own data comparing them with the content of a given database, both for population genetics and mitochondrial disease data. RESULTS: HmtDB is a well integrated web-based human mitochondrial bioinformatic resource aimed at supporting population genetics and mitochondrial disease studies, thanks to a new approach based on site-specific nucleotide and aminoacid variability estimation. HmtDB consists of a database of Human Mitochondrial Genomes, annotated with population data, and a set of bioinformatic tools, able to produce site-specific variability data and to automatically characterize newly sequenced human mitochondrial genomes. A query system for the retrieval of genomes and a web submission tool for the annotation of new genomes have been designed and will soon be implemented. The first release contains 1255 fully annotated human mitochondrial genomes. Nucleotide site-specific variability data and multialigned genomes can be downloaded. Intra-human and inter-species aminoacid variability data estimated on the 13 coding for proteins genes of the 1255 human genomes and 60 mammalian species are also available. HmtDB is freely available, upon registration, at http://www.hmdb.uniba.it. CONCLUSION: The HmtDB project will contribute towards completing and/or refining haplogroup classification and revealing the real pathogenic potential of mitochondrial mutations, on the basis of variability estimation. PMID- 16351754 TI - Query3d: a new method for high-throughput analysis of functional residues in protein structures. AB - BACKGROUND: The identification of local similarities between two protein structures can provide clues of a common function. Many different methods exist for searching for similar subsets of residues in proteins of known structure. However, the lack of functional and structural information on single residues, together with the low level of integration of this information in comparison methods, is a limitation that prevents these methods from being fully exploited in high-throughput analyses. RESULTS: Here we describe Query3d, a program that is both a structural DBMS (Database Management System) and a local comparison method. The method conserves a copy of all the residues of the Protein Data Bank annotated with a variety of functional and structural information. New annotations can be easily added from a variety of methods and known databases. The algorithm makes it possible to create complex queries based on the residues' function and then to compare only subsets of the selected residues. Functional information is also essential to speed up the comparison and the analysis of the results. CONCLUSION: With Query3d, users can easily obtain statistics on how many and which residues share certain properties in all proteins of known structure. At the same time, the method also finds their structural neighbours in the whole PDB. Programs and data can be accessed through the PdbFun web interface. PMID- 16351755 TI - MicroGen: a MIAME compliant web system for microarray experiment information and workflow management. AB - BACKGROUND: Improvements of bio-nano-technologies and biomolecular techniques have led to increasing production of high-throughput experimental data. Spotted cDNA microarray is one of the most diffuse technologies, used in single research laboratories and in biotechnology service facilities. Although they are routinely performed, spotted microarray experiments are complex procedures entailing several experimental steps and actors with different technical skills and roles. During an experiment, involved actors, who can also be located in a distance, need to access and share specific experiment information according to their roles. Furthermore, complete information describing all experimental steps must be orderly collected to allow subsequent correct interpretation of experimental results. RESULTS: We developed MicroGen, a web system for managing information and workflow in the production pipeline of spotted microarray experiments. It is constituted of a core multi-database system able to store all data completely characterizing different spotted microarray experiments according to the Minimum Information About Microarray Experiments (MIAME) standard, and of an intuitive and user-friendly web interface able to support the collaborative work required among multidisciplinary actors and roles involved in spotted microarray experiment production. MicroGen supports six types of user roles: the researcher who designs and requests the experiment, the spotting operator, the hybridisation operator, the image processing operator, the system administrator, and the generic public user who can access the unrestricted part of the system to get information about MicroGen services. CONCLUSION: MicroGen represents a MIAME compliant information system that enables managing workflow and supporting collaborative work in spotted microarray experiment production. PMID- 16351756 TI - An integrated approach of immunogenomics and bioinformatics to identify new Tumor Associated Antigens (TAA) for mammary cancer immunological prevention. AB - BACKGROUND: Neoplastic transformation is a multistep process in which distinct gene products of specific cell regulatory pathways are involved at each stage. Identification of overexpressed genes provides an unprecedented opportunity to address the immune system against antigens typical of defined stages of neoplastic transformation. HER-2/neu/ERBB2 (Her2) oncogene is a prototype of deregulated oncogenic protein kinase membrane receptors. Mice transgenic for rat Her2 (BALB-neuT mice) were studied to evaluate the stage in which vaccines can prevent the onset of Her2 driven mammary carcinomas. As Her2 is not overexpressed in all mammary carcinomas, definition of an additional set of tumor associated antigens (TAAs) expressed at defined stages by most breast carcinomas would allow a broader coverage of vaccination. To address this question, a meta-analysis was performed on two transcription profile studies to identify a set of new TAA targets to be used instead of or in conjunction with Her2. RESULTS: The five TAAs identified (Tes, Rcn2, Rnf4, Cradd, Galnt3) are those whose expression is linearly related to the tumor mass increase in BALB-neuT mammary glands. Moreover, they have a low expression in normal tissues and are generally expressed in human breast tumors, though at a lower level than Her2. CONCLUSION: Although the number of putative TAAs identified is limited, this pilot study suggests that meta-analysis of expression profiles produces results that could assist in the designing of pre-clinical immunopreventive vaccines. PMID- 16351757 TI - Secondary structure assignment that accurately reflects physical and evolutionary characteristics. AB - BACKGROUND: Secondary structure is used in hierarchical classification of protein structures, identification of protein features, such as helix caps and loops, for fold recognition, and as a precursor to ab initio structure prediction. There are several methods available for assigning secondary structure if the three dimensional structure of the protein is known. Unfortunately they differ in their definitions, particularly in the exact positions of the termini. Additionally, most existing methods rely on hydrogen bonding, which means that important secondary structural classes, such as isolated beta-strands and poly-proline helices cannot be identified as they do not have characteristic hydrogen-bonding patterns. For this reason we have developed a more accurate method for assigning secondary structure based on main chain geometry, which also allows a more comprehensive assignment of secondary structure. RESULTS: We define secondary structure based on a number of geometric parameters. Helices are defined based on whether they fit inside an imaginary cylinder: residues must be within the correct radius of a central axis. Different types of helices (alpha, 3(10) or pi) are assigned on the basis of the angle between successive peptide bonds. beta strands are assigned based on backbone dihedrals and with alternating peptide bonds. Thus hydrogen bonding is not required and beta-strands can be within a parallel sheet, antiparallel sheet, or can be isolated. Poly-proline helices are defined similarly, although with three-fold symmetry. CONCLUSION: We find that our method better assigns secondary structure than existing methods. Specifically, we find that comparing our methods with those of others, amino-acid trends at helix caps are stronger, secondary structural elements less likely to be concatenated together and secondary structure guided sequence alignment is improved. We conclude, therefore, that secondary structure assignments using our method better reflects physical and evolutionary characteristics of proteins. The program is available from http://www.bioinf.man.ac.uk/~lovell/segno.shtml. PMID- 16351758 TI - ParPEST: a pipeline for EST data analysis based on parallel computing. AB - BACKGROUND: Expressed Sequence Tags (ESTs) are short and error-prone DNA sequences generated from the 5' and 3' ends of randomly selected cDNA clones. They provide an important resource for comparative and functional genomic studies and, moreover, represent a reliable information for the annotation of genomic sequences. Because of the advances in biotechnologies, ESTs are daily determined in the form of large datasets. Therefore, suitable and efficient bioinformatic approaches are necessary to organize data related information content for further investigations. RESULTS: We implemented ParPEST (Parallel Processing of ESTs), a pipeline based on parallel computing for EST analysis. The results are organized in a suitable data warehouse to provide a starting point to mine expressed sequence datasets. The collected information is useful for investigations on data quality and on data information content, enriched also by a preliminary functional annotation. CONCLUSION: The pipeline presented here has been developed to perform an exhaustive and reliable analysis on EST data and to provide a curated set of information based on a relational database. Moreover, it is designed to reduce execution time of the specific steps required for a complete analysis using distributed processes and parallelized software. It is conceived to run on low requiring hardware components, to fulfill increasing demand, typical of the data used, and scalability at affordable costs. PMID- 16351759 TI - Monounsaturated fatty acids in parenteral nutrition; evaluation of risks and benefits. PMID- 16351760 TI - Duration of exclusive breast-feeding: introduction of complementary feeding may be necessary before 6 months of age. AB - The WHO recommends exclusive breast-feeding for the first 6 months of life. At present, <2 % of mothers who breast-feed in the UK do so exclusively for 6 months. We propose the testable hypothesis that this is because many mothers do not provide sufficient breast milk to feed a 6-month-old baby adequately. We review recent evidence on energy requirements during infancy, and energy transfer from mother to baby, and consider the adequacy of exclusive breast-feeding to age 6 months for mothers and babies in the developed world. Evidence from our recent systematic review suggests that mean metabolisable energy intake in exclusively breast-fed infants at 6 months is 2.2-2.4 MJ/d (525-574 kcal/d), and mean energy requirement approximately 2.6-2.7 MJ/d (632-649 kcal/d), leading to a gap between the energy provided by milk and energy needs by 6 months for many babies. Our hypothesis is consistent with other evidence, and with evolutionary considerations, and we briefly review this other evidence. The hypothesis would be testable in a longitudinal study of infant energy balance using stable-isotope techniques, which are both practical and valid. PMID- 16351761 TI - High concordance of daidzein-metabolizing phenotypes in individuals measured 1 to 3 years apart. AB - Particular intestinal bacteria are capable of metabolizing the soya isoflavone daidzein to equol and/or O-desmethylangolensin (O-DMA), and the presence of these metabolites in urine after soya consumption are markers of particular intestinal bacteria profiles. Prevalences of equol producers and O-DMA producers are approximately 30-50 % and 80-90 %, respectively, and limited observations have suggested that these daidzein-metabolizing phenotypes are stable within individuals over time. Characterizing stability of these phenotypes is important to understand their potential as markers of long-term exposure to particular intestinal bacteria and their associations with disease risk. We evaluated concordance within an individual for the equol-producer and O-DMA-producer phenotypes measured at two time points (T1, T2), 1-3 years apart. Phenotypes were ascertained by analysing equol and O-DMA using GC-MS in a spot urine sample collected after 3 d soya (source of daidzein) supplementation. In ninety-two individuals without recent (within 3 months before phenotyping) or current antibiotics use, 41 % were equol producers at T1 and 45 % were equol producers at T2, and 90 % were O-DMA producers at T1 and 95 % were O-DMA producers at T2. The percentage agreement for the equol-producer phenotype was 82 and for the O-DMA producer phenotype was 89. These results indicate that these phenotypes are stable in most individuals over time, suggesting that they provide a useful biomarker for evaluating disease risk associated with harbouring particular intestinal bacteria responsible for, or associated with, the metabolism of the soya isoflavone daidzein. PMID- 16351763 TI - Lactation decreases pancreatic lipase mRNA level in the rat. AB - Lactation alters maternal metabolism and increases food intake in rats to support milk production. Pancreatic lipase (PL) is primarily responsible for fat digestion in adults and is regulated by dietary fat. The present research determined the regulation of PL by lactation and dietary fat. In Expt 1, eighteen Sprague-Dawley dams and twelve age-matched virgins (controls) were fed a low-fat diet (LF; 11 % energy as safflower oil) for 7-63 d. At postpartum (day 0), peak lactation (day 15) and post-lactation (day 56) and after 7 d in virgins, the pancreas was removed for mRNA and enzyme analyses. In Expt 2, thirty-six Sprague Dawley dams were fed LF until day 9 postpartum when dams were divided into three groups of twelve; one continued to be fed LF, one was fed a moderate-fat diet (MF; 40 % energy as safflower oil); and one was fed a high-fat diet (HF; 67 % energy as safflower oil) diet. At peak lactation (day 15) and post-lactation (day 56), the pancreas was removed for mRNA and enzyme analyses. Expt 1 revealed that lactation and post-lactation significantly (P<0.001) decreased PL mRNA (67 % and 76 %, respectively), but only post-lactation decreased PL activity. Increased dietary fat in Expt 2 significantly increased PL mRNA (LF3, n 7) or low body condition score (LBCS; <2, n 6), thereby enabling us to examine the effect of maternal body composition on later cardiovascular function. From day 80 to term (approximately 147 d) all sheep were fed to 100 % ME. Offspring were weaned at 12 weeks and pasture-reared until 6 months of age when cardiovascular function was determined. Both LBCS and NR sheep tended to have lower resting systolic (control, 85 (se 2); LBCS, 77 (se 3); NR, 77 (se 3) mmHg) and diastolic blood pressure relative to controls. Total nephron count was markedly lower in both LBCS and NR relative to controls (LBCS, 59 (se 6); NR, 56 (se 12) %). Our data suggest that maternal body composition around conception is as important as the level of nutrient intake during early pregnancy in programming later cardiovascular health. PMID- 16351772 TI - Postprandial glycaemic, lipaemic and haemostatic responses to ingestion of rapidly and slowly digested starches in healthy young women. AB - The objective of the present study was to investigate the postprandial metabolism of two starches with contrasting rates of hydrolysis in vitro. Characterized using the Englyst method of in vitro starch classification, C*Set 06 598 contained predominantly rapidly digestible starch and C*Gel 04 201 contained predominantly slowly digestible starch. Each test starch, naturally enriched with 13C, was fed to ten healthy female volunteers as part of a moderate fat test meal (containing 75 g test starch and 21 g fat), in a double-blind randomized crossover design. The metabolic response to each starch was measured after an overnight fast, in an acute 6 h study, before and after 14 d of daily consumption of 75 g test starch. During each acute study, blood samples were taken at 15 min intervals for the first 2 h and at 30 min intervals for the remaining 4 h. Breath 13CO2 enrichment was measured at the same time points and indirect calorimetry was performed for 20 min every 40 min immediately before and throughout the study. Significantly more rapid, greater changes in postprandial plasma glucose, NEFA and serum insulin concentrations were observed after consumption of the rapidly digestible starch. Breath 13CO2 output over the first 3-4 h rose rapidly then began to decline following consumption of the rapidly digestible starch, but plateaued for the slowly digestible starch. The 14 d adaptation period did not affect any of the glycaemic or lipaemic variables but there was a reduction in postprandial plasminogen activator inhibitor-1 concentrations. These data confirm that starches characterized as predominantly rapidly digestible versus slowly digestible by the Englyst procedure provoke distinctly different patterns of metabolism postprandially. PMID- 16351773 TI - Effect of apolipoprotein E genotype on vitamin K status in healthy older adults from China and the UK. AB - The vitamin K concentration in the circulation and the availability of vitamin K to bone may be affected by factors influencing lipoprotein metabolism, such as apoE genotype. The relationships between markers of vitamin K status, bone mineral content and apoE genotype were studied in healthy older men and women aged 60-83 years, 177 from Shenyang, China, and 132 from Cambridge, UK. Fasting plasma was analysed for vitamin K1, triacylglycerol, total osteocalcin, undercarboxylated osteocalcin (ucOC) and apoE genotype. Hip bone mineral content was measured using dual-energy X-ray absorptiometry. Subjects were grouped according to apoE genotype as E2/3, E3/3 and [E3/4+E4/4]. The mean plasma vitamin K1 concentration of the three genotype groups was significantly higher and the percentage ucOC was lower in the Chinese than in the British subjects (P<0.01). A higher vitamin K1 concentration was found in subjects with [E3/4+E4/4] than those with either E2/3 or E3/3 in Cambridge (32.2 (SE 14.6 ) %, P=0.03; 24.6 (SE 10.7 ) %, P=0.02). Similar trends were observed although were not statistically significant in Shenyang (26.5 (18.9) %, P=0.16; 23.1 (13.0) %, P=0.08). Subjects with [E3/4+E4/4] had a lower percentage ucOC (total osteocalcin adjusted) than did those with either E2/3 or E3/3 in Shenyang (65.1 (27.2) %, P=0.02; 49.6 (19.9) %, P=0.01 respectively) but not in Cambridge. This study demonstrates that a superior vitamin K status is associated with the apoE4 genotype in healthy older individuals from China and the UK. PMID- 16351774 TI - The effect of selenium on thyroid status in a population with marginal selenium and iodine status. AB - The effects of Se on thyroid metabolism in a New Zealand population are investigated, including (a) the relationship between Se and thyroid status, and (b) the effect of Se supplementation on thyroid status. The data used come from two cross-sectional studies of Se, I, thyroid hormones and thyroid volume (studies 1 and 4), and three Se intervention studies in which thyroid hormones, Se and glutathione peroxidase (GPx) activities were measured (studies 2, 3 and 5). There were no significant correlations between Se status and measures of thyroid status after controlling for sex at baseline or after supplementation in any of the studies. When data from study 4 were divided into two groups according to plasma Se, plasma thyroxine (T4) was lower in males with higher plasma Se levels (P=0.009). Se supplementation increased plasma Se and GPx activity, but produced only small changes in plasma T4 and triiodothyronine (T3):T4 ratio. In study 2, there was a significant reduction in plasma T4 (P=0.0045). In studies 3 and 5 there were small decreases in plasma T4 and a small increase in the T3:T4 ratio, which were not significantly different from placebo groups. Lack of significant associations between plasma Se and thyroid status, and only small changes in T4 suggest that Se status in New Zealand is close to adequate for the optimal function of deiodinases. Adequate plasma Se may be approximately 0.82 0.90 micromol/l, compared with 1.00-1.14 micromol/l for maximal GPx activities. PMID- 16351776 TI - Resting energy expenditure measured longitudinally following hip fracture compared to predictive equations: is an injury adjustment required? AB - The present study measuring resting energy expenditure (REE; kJ/d) longitudinally using indirect calorimetry in six elderly women aged > or =70 years following surgery for hip fracture, describes changes over time (days 10, 42 and 84 post injury) and compares measured values to those calculated from routinely applied predictive equations. REE was compared to REE predicted using the Harris Benedict and Schofield equations, with and without accounting for the theoretical increase in energy expenditure of 35 % secondary to physiological stress of injury and surgery. Mean (95 % CI) measured REE (kJ/d) was 4704 (4354, 5054), 4090 (3719, 4461) and 4145 (3908, 4382) for days 10, 42 and 84, respectively. A time effect was observed for measured REE, P=0.003. Without adjusting for stress the mean difference and 95 % limits of agreement for measured and predicted REE (kJ/kg per d) for the Harris Benedict equation were 1 (-9, 12), 10 (2, 18) and 9 (1, 17) for days 10, 42 and 84, respectively. The mean difference and 95 % limits of agreement for measured and predicted REE (kJ/kg per d) for the Schofield equation without adjusting for stress were 8 (-3, 19), 16 (6, 26) and 16 (10, 22) for days 10, 42 and 84, respectively. After adjusting for stress, REE predicted from the Harris Benedict or Schofield equations overestimated measured REE by between 38 and 69 %. Energy expenditure following fracture is poorly understood. Our data suggest REE was relatively elevated early in recovery but declined during the first 6 weeks. Using the Harris Benedict or Schofield equations adjusted for stress may lead to overestimation of REE in the clinical setting. Further work is required to evaluate total energy expenditure before recommendations can be made to alter current practice for calculating theoretical total energy requirements of hip fracture patients. PMID- 16351775 TI - Haematological response to iron supplementation is reduced in children with asymptomatic Helicobacter pylori infection. AB - We evaluated the adverse effect of asymptomatic Helicobacter pylori infection in children on the response to Fe supplementation. One hundred and sixty-nine children aged 1-10 years from the urban poor community underwent a [13C]urea breath test for H. pylori and haematological tests at admission and after 8 weeks. Both H. pylori-positive and -negative children were randomly assigned to receive ferrous fumarate syrup (20 mg elemental Fe twice daily) or placebo for 8 weeks and a single dose of vitamin A (33,000 microg). Admission findings were compared between H. pylori-positive and -negative children. Response to Fe was compared between Fe-supplemented H. pylori-positive and -negative children. Seventy-nine per cent of the children were aged 1-5 years and half of them were boys. In eighty-five H. pylori-positive and eighty-four H. pylori-negative children, the differences in mean Hb (112 (sd 12.6) v. 113 (sd 12.0) g/l), haematocrit (34 (sd 3.5) v. 35 (sd 3.2) %) and ferritin (23.8 v. 21.0 microg/l) were similar. After 8 weeks of Fe supplementation, mean Hb was 5.3 g/l more (95 % CI 1.59, 9.0) and haematocrit was 1.4 % more (95 % CI 0.2, 2.6) in H. pylori negative (n 44) compared with H. pylori-positive (n 42) children. Mean ferritin was similar at admission and improved in both H. pylori-positive and -negative children. Asymptomatic H. pylori infection was not associated with higher rates of anaemia or Fe deficiency in children, but had a significant adverse effect on response to Fe therapy. However, this result is based on exploratory analysis and needs confirmation. PMID- 16351777 TI - Meat consumption reduces the risk of nutritional rickets and osteomalacia. AB - Endogenous vitamin D deficiency (low serum 25(OH)D3) is a necessary but insufficient requirement for the genesis of vitamin D-deficiency rickets and osteomalacia. The magnitude of the independent contributions of dietary factors to rachitic and osteomalacic risk remains uncertain. We reanalysed two weighed dietary surveys of sixty-two cases of rickets and osteomalacia and 113 normal women and children. The independent associations of four dietary variables (vitamin D, Ca, fibre and meat intakes) and daylight outdoor exposure with rachitic and osteomalacic relative risk were estimated by multivariate logistic regression. Meat and fibre intakes showed significant negative and positive associations respectively with rachitic and osteomalacic relative risk (RR; zero meat intake: RR 29.8 (95 % CI 4.96, 181), P<0.001; fibre intake: RR 1.53 (95 % CI 1.01, 2.32), P=0.043). The negative association of meat intakes with rachitic and osteomalacic relative risk was curvilinear; relative risk did not fall further at meat intakes above 60 g daily. Daylight outdoor exposure showed a significant negative association with combined relative risk (RR 0.33 (95 % CI 0.17, 0.66), P<0.001). Operation of the meat and fibre risk factors was related to sex, age and dietary pattern (omnivore/lactovegetarian), mainly determined by religious affiliation. The mechanism by which meat reduces rachitic and osteomalacic risk is uncertain and appears independent of revised estimates of meat vitamin D content. The meat content of the omnivore Western diet may explain its high degree of protection against nutritional rickets and osteomalacia from infancy to old age in the presence of endogenous vitamin D deficiency. PMID- 16351778 TI - Dietary glycaemic index and glycaemic load in Danish children in relation to body fatness. AB - The aim of this study was to describe dietary glycaemic index (GI) and glycaemic load (GL) values in the diets of Danish children, and to examine the associations between dietary GI, GL and body fatness. Data were collected during 1997-8 as part of the European Youth Heart Study. The study population comprised 485 children aged 10 years and 364 children aged 16 years from Odense County, Denmark. Dietary GI and GL were estimated using international food tables, and the associations between energy-adjusted dietary GI, GL and body fatness were analysed by multiple linear regression. The mean daily dietary GI value was 85 (SD 6.9) with a range of 62-111. No significant differences were found between age groups and gender. The daily dietary GL was higher among boys aged 16, with a GL of 330 (sd 95) (P<0.05), compared with girls or younger boys. Dietary GL was higher among 10-year-old boys than girls (250 (sd 81) v. 230 (sd 66) P<0.05), whereas dietary GL among 16-year-old girls was 230 (sd 56). Neither dietary GI nor GL was associated with the sum of four skinfolds (SigmaSF) among girls or among 10-year-old boys. Among 16-year-old boys, significant associations were observed between dietary GI and SigmaSF (beta=0.60, SE=0.21, P=0.006), and between dietary GL and SigmaSF (beta=0.15, SE=0.06, P=0.009). In conclusion, dietary GI and GL were positively associated with body fatness among Danish boys aged 16 years, whereas no associations were found among girls or younger boys. PMID- 16351779 TI - Validation of a pre-coded food diary with energy expenditure, comparison of under reporters v. acceptable reporters. AB - The objective of the present study was to compare energy intake (EI) assessed from a pre-coded food diary (PFD) with energy expenditure (EE) measured by a validated position-and-movement monitor (ActiReg); PreMed AS, Oslo, Norway) in a group of Norwegian 9-year-olds. Moreover, we examined whether and how under reporters (UR), identified with ActiReg), differed from acceptable reporters (AR) according to food intake and BMI. A total of fifty-one 9-year-olds completed PFD and ActiReg. The present study showed that on average EI was underestimated by 18 % compared with EE measured by ActiReg. The 95 % confidence limits of agreement in a Bland-Altman plot for EI and EE varied from 1.97 MJ to -4.23 MJ (sd 2) among the girls and from 0.74 MJ to -5.26 MJ (sd 2) among the boys. The Pearson correlation coefficient between EI and EE was 0.28 (P=0.05) for males and females combined. Fifty-seven per cent of the participants were classified as AR, 39 % as UR and 4 % as over-reporters with the PFD. Under-reporting of energy remains a problem with the PFD method used in a group of 9-year-olds, especially among boys. However, UR and AR did not show a systematic misreporting related to macronutrients, unhealthy foods or BMI. PMID- 16351780 TI - Arterial stiffness in 10-year-old children: current and early determinants. AB - It has been suggested that CVD has its origins in early life. An impairment of fetal growth and early postnatal nutrition may have programming effects on cardiovascular physiology. In addition, traditional risk factors for CVD may initiate the atherosclerotic process during childhood. We explored the effect of fat intake, physical activity and lipid profile in childhood, and birth weight, growth during infancy and breast-feeding on arterial stiffness in a cohort study of ninety-three 10-year-old children followed during infancy and re-examined at the age of 10 years. Arterial stiffness in two arterial segments (aorto-radial and aorto-femoral) was measured as pulse wave velocity. Arterial stiffness was inversely associated with physical activity (a regression coefficient in cm/s (95 % CI) of -6.8 (-11.2, -2.4) and -3.9 (-6.9, -0.8) per h of high physical activity/d in the aorto-radial and aorto-femoral segments, respectively). Arterial stiffness was also positively associated with dietary fat energy percentage (3.1 (95 % CI 0.9, 5.2) and 1.8 (95 % CI 0.2, 3.2) per fat energy percentage in the aorto-radial and aorto-femoral segments, respectively) but was not related to body composition, insulin resistance or lipid profile. Arterial stiffness was also positively associated with duration of breast-feeding for the aorto-femoral segment only (2.1 (95 % CI 0.4, 3.7) per month) but was not associated with growth in early life. In conclusion, patterns of physical activity and diet, and history of breast-feeding in infancy, have an influence on the stiffness of the large arteries in children. The long-term effects of this are unknown. PMID- 16351781 TI - Dietary fibre-rich oat-based products affect serum lipids, microbiota, formation of short-chain fatty acids and steroids in rats. AB - Wistar rats (ten per group) were fed either an oat-free control diet or a dietary fibre-rich test diet containing 500 g oat-based products/kg for 6 weeks. The oat based products, containing 4-128 g/kg resistant starch, 30-92 g/kg beta-glucan and 122-304 g/kg total dietary fibre, were oat flour extrudate, flour/Novelose (commercial resistant starch) extrudate (80:20 w/w), oat bran, bran/Novelose extrudate (80:20 w/w) and autoclaved oat flour. Serum total cholesterol decreased in the groups fed flour, flour/Novelose and bran/Novelose (P<0.05). In most of the test groups, count numbers of bifidobacteria were higher (P<0.001) and of coliforms were lower (P<0.05). The mass of the caecum walls and contents was greater in groups fed Novelose- and bran-containing diets (P<0.005). In all the test groups, pH values were lower in the intestinal contents (P<0.001), and caecal concentrations of acetate (P<0.001), propionate (P<0.05), butyrate (P<0.005) and total SCFA (P<0.001) were higher. The lowest concentrations of steroids were found in rats fed the autoclaved flour. In the other test groups, more bile acids appeared in the caecal (P<0.001) and colonic contents (P<0.005), as well as in the faeces, at week 6 (P<0.001). The highest bile acid excretion was found after feeding bran-containing diets. In the intestinal contents of all the test groups, more primary bile acids (P<0.001) appeared than in the control group. The excretion of steroids increased within the experimental period. Using extrusion technology, dietary fibre-rich oat-based products, which have beneficial physiological effects in rats, can be produced. Oat flour and bran are excellent sources for the preparation of directly edible oat products. Their nutritional properties can be further improved by the addition of resistant starch. PMID- 16351782 TI - Effect of green tea on resting energy expenditure and substrate oxidation during weight loss in overweight females. AB - We assessed the effect of ingestion of green tea (GT) extract along with a low energy diet (LED) on resting energy expenditure (REE), substrate oxidation and body weight as GT has been shown to increase energy expenditure and fat oxidation in the short term in both animals and people. Forty-six overweight women (BMI 27.6 (sd 1.8) kg/m2) were fed in energy balance from day 1 to day 3, followed by a LED with GT (1125 mg tea catechins +225 mg caffeine/d) or placebo (PLAC) from day 4 to day 87. Caffeine intake was standardised to 300 mg/d. Energy expenditure was measured on days 4 and 32. Reductions in weight (4.19 (sd 2.0) kg PLAC, 4.21 (sd 2.7) kg GT), BMI, waist:hip ratio, fat mass and fat-free mass were not statistically different between treatments. REE as a function of fat-free mass and fat mass was significantly reduced over 32 d in the PLAC group (P<0.05) but not in the GT group. Dietary restraint increased over time (P<0.001) in both groups, whereas disinhibition and general hunger decreased (P<0.05). The GT group became more hungry over time and less thirsty, and showed increased prospective food consumption compared with PLAC (P<0.05). Taken together, the ingestion of GT along with a LED had no additional benefit for any measures of body weight or body composition. Although the decrease in REE as a function of fat-free mass and fat mass was not significant with GT treatment, whereas it was with PLAC treatment, no significant effect of treatment over time was seen, suggesting that a robust limitation of REE reduction during a LED was not achieved by GT. PMID- 16351783 TI - Glycaemic index of potatoes: myth and reality from a European perspective. PMID- 16351784 TI - Effects of school-milk intervention on growth and bone mineral accretion in Chinese girls aged 10-12 years: accounting for cluster randomisation. PMID- 16351785 TI - [Comparison between R-CHOP regimen and CHOP regimen in treating naive diffuse large B-cell lymphoma in China--a multi-center randomized trail]. AB - BACKGROUND & OBJECTIVE: CHOP regimen is a standard treatment for patients with diffuse large B-cell non-Hodgkin's lymphoma (NHL), and its 5-year overall survival (OS) rate is 30%-40%. Rituximab is a chimeric monoclonal antibody (MoAb) directly against CD20-positive B cells, and has good effect on diffuse large B cell NHL. Rituximab combined with standard chemotherapy has been approved for treating aggressive B-cell NHL in Europe and the US. This study was to determine efficacy and safety of the combination of Rituximab and CHOP regimen in treating Chinese patients with CD20-positive diffuse large B-cell NHL. METHODS: From Sep. 2003 to Nov. 2004, a total of 63 patients in 9 centers were enrolled. All the patients were randomized into 2 groups: 32 received CHOP regimen alone (CHOP group), and 31 received Rituximab and CHOP regimen (R-CHOP group). All patients signed informed consent. The complete response rates, overall response rates, and side events of the 2 groups were compared. RESULTS: The complete response rates were similar in R-CHOP and CHOP groups (41.9% vs. 37.5%, P=0.719); the overall response rates were slightly higher in R-CHOP group than in CHOP group (83.8% vs. 65.6%, P=0.096). Disease progression during treatment was reported for 7 (21.9%) patients in CHOP group and 1 (3.2%) patient in R-CHOP group (P=0.026). The occurrence rates of adverse events were similar in R-CHOP and CHOP groups (65.6% vs. 67.7%, P=0.859). The most common adverse event was leukopenia; fever and chills were rather common in R-CHOP group. Clinically relevant toxicity was similar in both groups. CONCLUSION: When compared with standard CHOP alone, the addition of Rituximab to standard CHOP regimen reduces the risk of treatment failure in patients with diffuse large B-cell NHL, and doesn't increase the occurrence of chemotherapy-related adverse events. PMID- 16351786 TI - [Effect of exogenous expression of heparanase gene on invasive ability of colorectal cancer cell line HT29]. AB - BACKGROUND & OBJECTIVE: Heparanase (Hpa) is an endoglycosidase that degrades heparin sulfate--the main polysaccharide constituent of extracellular matrix (ECM) and basement. It can enhance the invasive and metastatic potential of malignant tumors and cell lines by destroying ECM and basement. This study was to explore the effects of heparanase gene on the invasive ability of colorectal cancer cell line HT29. METHODS: Heparanase gene was transfected into HT29 cells. Cell growth kinetics was assessed by MTT assay, and in vitro invasive ability was assessed with Boyden chamber. Xenograft and orthotopic implantation of histologically intact tumor in nude mice were constructed to observe the effect of exogenous expression of heparanase gene on invasive ability of HT29 cells. RESULTS: After transfection, the growth rate of heparanase-transfected HT29 (HT29 Hpa) cells was obviously higher than those of untransfected HT29 cells and empty vector-transfected HT29 (HT29-KZ) cells; the invasive cell number was significantly larger in HT29-Hpa cells than in untransfected HT29 cells and HT29 KZ cells (45.5+/-0.5 vs. 29.3+/-0.1 and 30.1+/-0.2, P<0.01). The entity neoplasm formed by HT29-Hpa cells (12 mm x 9 mm x 10 mm) was bigger than that formed by untransfected HT29 cells (6 mm x 8 mm x 6 mm). The prevalence of liver metastasis caused by orthotopic implantation of xenograft was significantly higher in HT29 Hpa group than in control group (71.43% vs. 14.29%, P<0.01). CONCLUSION: Exogenous heparanase gene can facilitate the growth, invasion, and metastasis of HT29 cells. PMID- 16351787 TI - [Inhibitory effect of silencing heparanase expression with antisense oligodeoxynucleotide on invasive capability of human esophageal squamous cancer cell line TE-13]. AB - BACKGROUND & OBJECTIVE: Heparanase, a kind of endo-D-glucuronidase, degrades heparin sulfate proteoglycans, and plays important roles in invasion and metastasis of many kinds of malignant tumors. This research was designed to investigate the expression of heparanase in esophageal squamous cancer cell line TE-13 and the effect of heparanase antisense oligodeoxynucleotide (ASODN) transfection on invasion of TE-13 cells. METHODS: Synthesized heparanase ASODN was transfected into TE-13 cells. Before and after transfection, the expression of heparanase mRNA and protein in TE-13 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunocytochemistry. Matrigel invasion assay was used to determine the invasive capability of TE-13 cells. RESULTS: In TE-13 cells, heparanase gene (585 bp) was detected by RT-PCR, and heparanase protein (50 ku) by Western blot. Heparanase protein mainly located in cytoplasm and on cell membrane. After heparanase ASODN transfection, heparanase gene and protein expression and the invasive TE-13 cells were significantly reduced along with the ascending concentration of heparanase ASODN (P<0.05, P<0.05). CONCLUSIONS: Heparanase gene is expressed in TE-13 cells. Heparanase ASODN can obviously inhibit heparanase gene expression, and restrain invasive activity of TE-13 cells. PMID- 16351788 TI - [Regulatory effect of small interfering RNA targeting multidrug resistant protein 1 on chemosensitivity of human multiforme glioblastoma cell line BT325]. AB - BACKGROUND & OBJECTIVE: Multidrug resistance is a major reason of failure of chemotherapy for glioma. Overexpression of P-glycoprotein (P-gp), encoded by multidrug resistant protein 1 (MDR1) gene, is one of the key factors. This study was to explore the regulatory effect of small interfering RNA (siRNA) targeting MDR1 on chemosensitivity of human multiforme glioblastoma cell line BT325. METHODS: MDR1 siRNAs containing sequences of 3,051-3,069 (MDR1 A group), 502-520 (MDR1 B group), and 1,534-1,552 (MDR1 C group) were designed, and transfected into BT325 cells. Positive clones were screened with puromycin. The expression of MDR1 was measured by reverse transcription-polymerase chain reaction (RT-PCR); the expression of P-gp was detected by immunohistochemistry and flow cytometry (FCM). Drug sensitivity assay was performed in the transfected cells. RESULTS: BT325 cells proliferated exponentially after MDR1 siRNA transfection. After transfection of MDR1 siRNAs, the expression of MDR1 mRNA was significantly lower in MDR1 A, B, and C groups than in control group (0.18+/-0.05, 0.30+/-0.09, and 0.36+/-0.13 vs. 0.76+/-0.06, P<0.001); the positive rate of P-gp was decreased from 85.73% to 1.44%; the 50% inhibitory concentrations (IC(50)) of doxorubicin and vincristine to BT325 cells were decreased markedly; the G0/G1 phase proportions of MDR1 A, B, and C groups were increased by 13.55%, 14.35%, and 1.46% of control, respectively (P<0.05). CONCLUSION: MDR1 siRNA may modulate multidrug resistance through down-regulating the expression of MDR1 gene, enhancing the chemosensitivity of glioma, and inducing cell apoptosis. PMID- 16351789 TI - [In vitro inhibitory effect of dibenzoxanthene CY-B12 on proliferation of tumor cell lines and its possible mechanisms]. AB - BACKGROUND & OBJECTIVE: CY-B12, a new dibenzon-xanthene, has been synthesized recently. This study was to investigate the in vitro antiproliferative activity of CY-B12 and its possible mechanisms. METHODS: The inhibitory effects of CY-B12 on proliferation of gastric carcinoma cell line MGC803, nasopharyngeal carcinoma cell line CNE-2, oral epithelial carcinoma cell line KB-3-1, and lung cancer cell line Glc82 were assessed by MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometry (FCM). Apoptotic morphology of CNE-2 cells was observed under fluorescent microscope after Hoechst33258 staining. The expression of cell cycle-related proteins Cdc25C, Cdc2, and Cyclin B1 were measured by Western blot. DNA damage was detected by single cell gel electrophoresis (SCGE). RESULTS: CY-B12 obviously inhibited the proliferation of MGC803, CNE-2, KB-3-1, and Glc82 cells; the IC(50) values were 7.51, 9.58, 8.84, and 15.99 micromol/L, respectively. After treatment of CY-B12, CNE-2 cells were arrested at G(2)/M phase; chromatin condensation, apoptotic bodies, and sub-G1 peak were observed in CNE-2 cells. The protein expression of Cdc25C in CNE-2 cells was down-regulated by CY-B12 in a dose-dependent manneru whereas Cyclin B1 and Cdc2 were up regulated by low dose of CY-B12, and down-regulated by high dose of CY-B12. CY B12 induced DNA damage in CNE-2 cells in a dose-dependent manner. CONCLUSION: CY B12 has potent in vitro antiproliferative activity, which may be exerted through breaking DNA, down-regulating cell cycle-related protein Cdc25C, and inducing cell cycle arrest and apoptosis. PMID- 16351790 TI - [Inhibitory effect of fumagillol combined with cyclophosphamide on metastasis of lung adenocarcinoma cell line LA795 xenograft in mice]. AB - BACKGROUND & OBJECTIVE: Treating tumor with angiogenesis inhibitor and chemotherapeutic drugs is a research hot spot now. This study was designed to observe the synergetic inhibitory effect of fumagillol (TNP-470) in combination with cyclophosphamide (CTX) on metastasis of lung adenocarcinoma cell line LA795 xenograft in mouse, and to explore the related mechanism of suppressing tumor metastasis by TNP-470. METHODS: Forty T739 nude mice bearing highly metastatic LA795 cells were randomized into 5 groups: control group, vehicle group, TNP-470 (30 mg/kg) group, CTX (40 mg/kg) group, and combination group (TNP-470 plus CTX). All mice were killed 3 weeks lateru the subcutaneous tumors were weighted to calculate inhibitory rate. The metastatic tumor foci on lung surface in mice were counted to calculate occurrence rate and inhibitory rate of metastases on lung surface. The microvessel density (MVD) and the expression of tumor metastasis related factor P-selectin in subcutaneous tumor were detected by immunohistochemistry and analyzed with image analysis system. RESULTS: The inhibitory rate of tumor was significantly higher in combination group (81.5%) than in other groups (P<0.01). TNP-470 plus CTX showed synergetic effect on inhibiting metastasis on lung surface with a Q value of 1.21. The metastatic foci on lung surface were significantly fewer in combination group, TNP-470 group, and CTX group than in control group (1.75+/-1.71, 4.75+/-3.34, and 8.50+/-2.67 vs. 12.13+/-4.02, P<0.05). The MVD and the expression of P-selectin in subcutaneous tumor were also significantly lower in combination group and TNP-470 group than in control group (9.13+/-1.61 and 12.13+/-2.84 vs. 20.50+/-3.12, P<0.01; 5.25+/ 2.27 and 7.13+/-3.01 vs. 13.75+/-3.38, P<0.01). CONCLUSIONS: TNP-470 and CTX have synergetic inhibitory effect on lung metastasis of LA795 xenograft tumor. TNP-470 may inhibit lung metastasis of LA795 xenograft tumor by suppressing the expression of P-selectin. PMID- 16351791 TI - [Antitumor effect and mechanism of shikonin derivative SYUNZ-7]. AB - BACKGROUND & OBJECTIVE: Natural shikonin compounds and their derivatives have cytotoxicity and antitumor effects. This study was to explore in vitro and in vivo antitumor effects of SYUNZ-7 [2 or 3, 11-bis(phenylsulfanyl)-6 isohexenylnaphthazarin] and the mechanisms. METHODS: In vitro antiproliferation effects of SYUNZ-7 on human lung adenocarcinoma cell line GLC-82, human nasopharyngeal cancer cell line CNE2, human oral cavity cancer cell line KB, human gastric cancer cell line MGC-803 and human hepatocellular cancer cell line HepG2 were tested by MTT assay. In vivo antitumor effect of SYUNZ-7 was tested using ascitic cancer EAC xenograft in mice and CNE2 xenograft in nude mice models. Cell apoptosis and cell cycle distribution were assessed by flow cytometry. The in vivo effect of SYUNZ-7 on angiogenesis was detected by immunohistochemistry. RESULTS: The 50% inhibitory concentrations (IC(50)) of SYUNZ-7 to GLC-82, CNE2, KB, MGC-803, and HepG2 cells were (2.18+/-0.04) microg/ml, (4.17+/-0.09) microg/ml, (5.41+/-0.10) microg/ml, (6.41+/-0.14) microg/ml, and (9.99+/-0.21) microg/ml, respectively. Under the treatment of 1, 2, 4, and 8 mg/kg of SYUNZ-7, the inhibitory rates of EAC xenografts in mice were (40.5+/-0.14)%, (50.9+/-2.3)%, (61.7+/-1.8)%, and (65.6+/-7.4)%, respectively (P<0.01). Under the treatment of 1, 2, and 4 mg/kg of SYUNZ-7, the inhibitory rates of CNE2 xenografts in nude mice were 24.7%, 38.3%, and 41.2%, respectively (P<0.05). SYUNZ-7 induced apoptosis of CNE2 cells in time- and concentration dependent manners, and blocked the transition of CNE2 cells from S to G(2)/M phase. SYUNZ-7 also inhibited the angiogenesis of CNE2 xenografts in nude mice in a concentration-dependent manner. CONCLUSION: SYUNZ-7 has strong in vivo and in vitro antitumor effects which are related to inducing cell apoptosis, blocking cell cycle, and inhibiting angiogenesis of tumor. PMID- 16351792 TI - [Inhibitory effects of Scutellaria barbatae D. Don on tumor angiogenesis and its mechanism]. AB - BACKGROUND & OBJECTIVE: Various chemically synthetic anti-angiogenesis agents have serious side effects. The traditional Chinese medicine has attracted considerable attention because of its low toxicity. This study was to explore the inhibitory effects of Scutellaria barbatae D. Don, a kind of traditional Chinese medicinal anti-cancer herb, on tumor angiogenesis, and investigate its mechanism. METHODS: Matrigel plug and human umbilical vascular endothelial cells (HUVECs) were used to construct in vivo and in vitro models of angiogenesis to assess the effect of Scutellaria barbatae D. Don on angiogenesis. After cultured with Scutellaria barbatae D. Don, the migration of endothelial cells was examined by Transwell chamber; the expression of vascular endothelial growth factor (VEGF) in HeLa cells was detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Scutellaria barbatae D. Don significantly inhibited angiogenesis in Matrigel; the tube formation number was significantly lower in 20% and 40% medicated serum groups containing Scutellaria barbatae D. Don than in 20% and 40% drug-free serum groups (5.6+/-1.1 vs. 9.8+/-1.3, P=0.001; 1.0+/-0.7 vs. 13.4+/-1.1, P<0.001). Migrated endothelial cells was significantly fewer in 20% and 40% medicated serum groups containing Scutellaria barbatae D. Don than in 20% and 40% drug-free serum groups (19.75+/-2.63 vs. 24.25+/-2.06, P=0.038; 14.00+/-2.58 vs. 26.5+/-4.65, P=0.006). When treated for 24 h and 48 h, the expression of VEGF in HeLa cells was significantly lower in 40% medicated serum group containing Scutellaria barbatae D. Don than in 40% drug-free serum group (138.67+/-9.50 vs. 195.82+/ 2.43, P=0.006; 93.84+/-41.11 vs. 193.68+/-18.37, P=0.036). CONCLUSION: Scutellaria barbatae D. Don could efficiently inhibit angiogenesis in tumor tissue which might relate with inhibition of endothelial cell migration and down regulation of VEGF in tumor cells. PMID- 16351793 TI - [Inducement effect of Meisoindigo on apoptosis of leukemia cell line HL-60 and its mechanism]. AB - BACKGROUND & OBJECTIVE: Meisoindigo is a powerful drug used in treating chronic myeloid leukemia (CML), but little is known about the mechanisms. This study was to investigate the inducement effect of meisoindigo on apoptosis of myelocytic leukemia cell line HL-60, and explore the possible mechanisms. METHODS: After treatment of meisoindigo, the proliferation of HL-60 cells was detected by trypan blue exclusion assay, and DNA fragmentation by agarose electrophoresis; cell morphology was observed under fluorescent microscope. Cell apoptosis and the expression of Fas were detected by flow cytometry. The expression of Caspase-3, Caspase-8, Caspase-9, PARP, Bcl-2, Bax and the concentration of cytochrome c in cytosol were analyzed by Western blot. RESULTS: Meisoindigo inhibited proliferation and induced apoptosis in HL-60 cells. When treated with 20 micromol/L meisoindigo for 12-48 h, the proliferation of HL-60 cells was significantly inhibited. When treated for 1 h, the apoptosis rate of HL-60 cells was (3.70+/-0.56)%; the apoptosis rate was significantly higher in HL-60 cells treated for 3, 6, and 12 h than in control cells [(19.80+/-1.13)%, (29.20+/ 2.69)%, and (47.05+/-7.70)% vs. (2.65+/-0.78)%, P<0.05]. When treated with meisoindigo for 3 h, typical changes of apoptosis, such as chromatin condensation and DNA ladder, were detected in HL-60 cells. The positive rate of Fas was significantly higher in cells treated with 20 micromol/L meisoindigo for 1 h than in control cells [(21.30+/-1.27)% vs. (9.35+/-0.21)%, P<0.05]. Meisoindigo activated Caspase-3, Caspase-8, Caspase-9 and PARP, down-regulated the expression of Bcl-2, up-regulated the expression of Bax and the concentration of cytochrome c. Furthermore, pretreatment of caspase-3 inhibitor z-DEVD-fmk partially reversed the inhibitory effect of meisoindigo on cell proliferation, and decreased cell apoptosis; when treated with meisoindigo for 5 h, the apoptosis rate was significantly higher in pretreated cells than in cells without pretreatment [(29.8+/-5.4)% vs. (16.5+/-5.5)%, P<0.05]; when treated with meisoindigo for 12 h, the alive cell number was significantly lower in pretreated cells than in cells without pretreatment [(1.80+/-0.14) x 10(5)/ml vs. (3.57+/-0.18) x 10(5)/ml, P<0.05]. CONCLUSION: Meisoindigo induces apoptosis of HL-60 cells which may relate to regulation of caspases pathway and bcl-2 family proteins. PMID- 16351794 TI - [Therapeutic effect of Seaweed Polysaccharide from Sargassum confusum on sarcoma s180 in mice and its mechanism]. AB - BACKGROUND & OBJECTIVE: Previous chemotherapeutic drugs have side effects which restrict their application in treating tumors. This research was to explore the inhibitory effect of wild Seaweed Polysaccharide from Sargassum confusum (SP) on sarcoma S180 xenograft in mice and its mechanism to offer experimental basis for searching for antitumor drugs with high efficiency and low toxicity. METHODS: SP were prepared and injected abdominally in mice at the doses of 50, 100, and 200 mg/kg, respectively, for 10 days before tumor cell transplantation. The mice with injection of 0.85% normal saline were set as negative control. The weight of S180 xenografts in mice was measured to calculate the inhibitory rate of SP on S180 tumor; the cell cycle of thymocytes and splenocytes were measured by flow cytometry (FCM); DNA and protein syntheses in thymocytes and splenocytes were measured by incorporation of (3)H-TdR; the immunopotentiative effect of SP on mice was assessed by splenic lymphocytes transformation assay. RESULTS: The inhibitory rates of S180 xenografts in mice treated with 50, 100, and 200 mg/kg SP were 28.7%, 32.9%, and 50.3%, respectively. The thymocytes and splenocytes functions were significantly enhanced in SP groups than in control group (P<0.05, P<0.01). CONCLUSIONS: SP might inhibit the growth of S180 xenograft in mice through improving the functions of thymocytes and splenocytes. PMID- 16351795 TI - [Correlation of antitumor effect of recombinant sea snake basic phospholipase A2 to its enzymatic activity]. AB - BACKGROUND & OBJECTIVE: Snake venom phospolipase A2 (PLA(2)), a large family of homologous (14 ku) soluble proteins, exerts diverse pharmacologic activities as well as enzymatic activities. So far, the structure and function of terrestrial snake PLA(2), especially the relationship of its enzymatic and pharmacologic activities have been studied extensively, but the investigation of sea snake PLA(2) are limited. This study was to investigate the in vitro and in vivo antitumor effects of recombinant sea snake basic PLA(2) (rSSBPLA(2)) and its mutants rN48 and rK4 from sea snake Lapemis hardwickii venom, and to explore the influence of 2 residues related with the enzymatic activity on the antitumor effects. METHODS: Site-directed mutagenesis of the 2 conserved residues related with enzymatic activity (His48 mutated to Asn and Asp49 mutated to Lys) was performed. The inhibitory effects of rSSBPLA(2), rN48 and rK49 on proliferation of human myeloid leukemia cell line HL-60, human neuroblastoma cell line SK-N-SH, human gastric cancer cell line MGC-803, and human liver cancer cell line HepG2 were assessed by MTT assay. Their antitumor effects on sarcoma cell line S180 xenograft and EAC ascites cancer model in mice were detected. RESULTS: The relative enzymatic activities of rN48 and rK49 were 0 and 5% of that of rSSBPLA(2). The 50% inhibitory concentration (IC(50)) of rSSBPLA(2) for HL60, SK N-SH, and MGC-803 cells were (45.28+/-0.09) microg/ml, (57.07+/-0.12) microg/ml, and (69.34+/-0.35) microg/ml, respectively, but it had no inhibitory effect on proliferation of HepG2 cells. rSSBPLA(2) obviously inhibited growth of S180 xenograft in miceu the inhibitory rates were 50.8%, 43.2%, 38.2%, and 55.5%, respectively, under the dose of 2 mg/kg (qd x 10), 2 mg/kg (q2d x 5), 4 mg/kg (qd x 1) and 4 mg/kg (q5d x 2). The inhibitory rate of EAC model was 33.5% under the dose of 4 mg/kg (q5d x 2). The inhibitory rates were significantly higher in test groups than in control groups (P<0.01). rN48 and rK49 had no inhibitory effect on proliferation of the 4 tumor cell lines and on growth of the xenograft tumors. CONCLUSION: The antitumor effect of rSSBPLA(2) may be closely related with its enzymatic activity. PMID- 16351796 TI - [Effect of caffeine on camptothecin-induced apoptosis of Molt-4 cells]. AB - BACKGROUND & OBJECTIVE: Caffeine could act on cell cycle checkpoints and affect the progression of cell cycle, but its impact on apoptosis of tumor cells is in debate. This study was carried out to investigate the effects of caffeine on camptothecin-induced apoptosis and cell cycle checkpoints of leukemia cell line Molt-4. METHODS: The cell apoptosis was induced by camptothecin, and caffeine was used to interfere with cell cycle checkpoints. The apoptosis rate and cell cycle during apoptosis were analyzed using sub-G1 method and Annexin V-propidium iodide (Annexin V/PI) staining. RESULTS: Caffeine (2.0-20.0 mmon/L) had no effect on proliferation of Molt-4 cells in exponentially growth phase. Camptothecin selectively induced apoptosis of Molt-4 cells in S phase; when induced with camptothecin (0.15 micromol/L) for 4 or 6 h, the apoptosis rates were (23.69+/ 2.26)% and (36.99+/-1.42)%. This cell cycle-specific apoptosis were inhibited obviously by caffeine with the apoptosis rates of (4.79+/-0.64)% and (2.69+/ 0.56)%. When caffeine was removed, the apoptosis rates increased obviously to (46.23+/-0.21)% and (55.81+/-0.41)%, and still mainly happened in S phase. CONCLUSIONS: Caffeine could inhibit camptothecin-induced apoptosis of Molt-4 cells. As a drug acting on cell cycle checkpoints, caffeine could transiently shield the surveillance of checkpoints to damaged cells and inhibit cell apoptosis. The effect may be reversed when caffeine is removed away. PMID- 16351797 TI - [Impact of CPP-Id on expression of co-stimulating molecules on dendritic cells in mice]. AB - BACKGROUND & OBJECTIVE: Idiotypic immunoglobin (Id) derived from B-cell lymphoma, as a tumor-specific antigen, can suppress tumor development by inducing immune response. This study was to confirm the presence of Id epitope on cytotoxic T lymphocytes (Id-CTL) in mouse lymphoma cell line A20, detect the quantity and distribution of cell-penetrating peptide-loaded Id (CPP-Id) in dendritic cells (DCs) at different time points, and explore its impact on the expression of surface molecules on DCs. METHODS: Id-CTL epitope in A20 cells was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. The quantity of CPP-Id and Id alone in DCs and the expression of DC surface molecules were detected by flow cytometry. The process of CPP-Id entering DCs was observed under confocal microscope, and its distribution was observed under fluorescent microscope. RESULTS: A 660 bp-length fragment was amplified from A20 cells by RT PCR, and identified as Id-CTL epitope by sequencing. CPP-Id entered DCs quickly during the initial 200 s, but few Id entered DCs. The mean fluorescent intensity (MFI) in DCs was significantly higher in CPP-Id group than in Id group (4.35+/ 0.48 vs. 1.14+/-0.33, P<0.005). The expression of surface molecules CD80, CD86, CD54 and MHC-I, MHC-II were higher on the DCs loaded by either CPP-Id or Id alone than on immature DCs. CONCLUSIONS: CPP can enhance the quantity of Id entering DCs. CPP-Id can up-regulate the expression of MHC I, MHC-II and CD80, CD86, and CD54 on DCs, and may help to activate CTLs. PMID- 16351798 TI - [Phase II randomized clinical trail of zoledronic acid in treating metastatic bone pain of malignancy]. AB - BACKGROUND & OBJECTIVE: Bone metastases are the prevailing reason for pain in patients with malignancies, which seriously affect their life quality. Zoledronic acid, the third generation of bisphosphonates, can inhibit the activity of osteoclasts and relieve pain. This study was to evaluate the efficacy and safety of zoledronic acid in treating metastatic bone pain for patients with malignancies. METHODS: A multi-center, randomized, double-blind, and prospective trail was conducted. Patients with metastatic bone pain were randomized to receive zoledromic acid plus monnitol, or pamidronate plus monnitol as positive control. RESULTS: From Oct. 2003 to Oct. 2004, 216 patients with metastatic bone pain were randomized into 2 groups: 109 in zoledromic acid group, and 107 in pamidronate group. There was no significant difference in pain intensity (PI) between zoledromic acid group and pamidronate group before treatment (6.0+/-1.1 vs. 6.0+/-1.3, P=0.938), 7 days after treatment (3.7+/-1.99 vs. 4.1+/-2.0, P=0.119), and 14 days after treatment (3.2+/-2.0 vs. 3.7+/-2.4, P=0.129). The differences in complete response (CR) rate, partial response (PR) rate, and total response rate between the 2 groups were not significant (10.4% vs. 9.5%, 69.8% vs. 69.5%, 88.7% vs. 85.7%, P>0.05). Time to CR was significantly shorter in zoledromic acid group than in pamidronate group [(7.0+/-2.2) days vs. (9.5+/-2.6) days, P=0.033]; the differences in time to PR, duration of CR, and duration of PR between the 2 groups were not significant [(4.9+/-2.6) days vs. (5.0+/-2.5) days, P=0.908; (13.2+/-1.80) days vs. (14.0+/-0.0) days, P=0.155; and (13.4+/-1.9) days vs. (12.8+/-2.8) days, P=0.127]. The main adverse events were fever, nausea, vomiting, and general malaise. The occurrence and severity of adverse events were similar between the 2 groups. CONCLUSION: Zoledromic acid is effective and safe in treating metastatic bone pain of patients with malignancy, which is similar to pamidranate. PMID- 16351799 TI - [Prognostic analysis of patients with nasal-type NK/T-cell non-Hodgkin's lymphoma -a report of 93 cases]. AB - BACKGROUND & OBJECTIVE: Nasal-type NK/T-cell non-Hodgkin's lymphoma (NHL) is a unique subtype with the manifestation of local necrosis, infection and fever. The efficacy of chemotherapy alone is unsatisfactory; while radiochemotherapy plays some roles in the management of NK/T-cell lymphoma (NK/TCL). This study was to summarize the clinical characteristics, treatment outcome and prognosis of NK/TCL patients. METHODS: Records of 93 patients with NK/TCL from Jan. 1997 to Jun. 2004 were analyzed retrospectively. All the patients were classified according to WHO classification system. RESULTS: Of the 93 patients, 75 (80.6%) were in stage I II, and 18 (19.4%) were in stage III-IV. The most common symptoms were nasal obstruction, rhinorrhea, and epistaxis. The disease course was 1-24 months with a median of 6.5 months. Of the 93 patients, 15 (16.1%) presented perforation of hard palate and/or nasal septum, 35 (37.6%) presented B symptoms; 35 (37.6%) were treated with chemotherapy alone, 2 (2.2%) were treated with radiotherapy alone, 54 (58.0%) were treated with radiochemotherapy, and 2 (2.2%) received no treatment. The first-line chemotherapy regimens were mainly CHOP and EPOCH. The overall response rate (RR) was 84.4% (76/90) with complete remission (CR) rate of 64.4% (58/90). The response rate of chemotherapy alone group was 67.6% (23/34) with CR rate of 41.2% (14/34). The response rate of combined modality group was 94.4% (51/54) with CR rate of 83.3% (45/54). The 2 patients who received no treatment died within 6 months. The major toxicity of chemotherapy was myelosuppression. The prevalence of grade III-IV neutropenia, thrombocytopenia, and anemia were 37.7%, 13.7%, and 10.7%. The major toxicities of radiotherapy were grade I-II mucosa lesion and myelosuppression. Other toxicities were mild. The mortality was 66.7% (62/93). The 1-, 3-, and 5-year overall survival (OS) rates were 63.4%, 43.1%, and 17.6%, respectively. Multivariate analysis showed that perforation of hard palate and/or nasal septum, B symptoms and therapeutic modality were independent prognostic factors of NK/TCL (P=0.035, P<0.001, and P=0.004). CONCLUSIONS: NK/TCL has low chemotherapy sensitivity. Although combined chemoradiotherapy yield better outcome, the long-term survival was still poor. Investigation of optional treatment is needed. PMID- 16351800 TI - [Comparison of rituximab plus CHOP regimen and CHOP regimen alone for treatment of newly diagnosed patients with diffuse large B-cell lymphoma]. AB - BACKGROUND & OBJECTIVE: CHOP regimen is the standard treatment for patients with diffuse large B-cell lymphoma. Rituximab, an anti-CD20 monoclonal antibody, is effective in treating diffuse large B-cell lymphoma. This study was conducted to compare the efficacy of rituximab plus CHOP and CHOP alone on newly diagnosed patients with diffuse large B-cell lymphoma, and analyze their toxicities. METHODS: A total of 72 newly diagnosed patients with diffuse large B-cell lymphoma were divided into 2 groups prospectively with concurrent control: 34 received CHOP plus rituximab (375 mg/m2, 2 days before each course) (combination group), 38 received CHOP alone. Each course lasted 3 weeks. All cases were evaluated after 6 courses. RESULTS: The total response rate in combination group was 93.8% (30/32), among which complete remission was seen in 23 patients and partial remission was seen in 7 patientsu while the total response rate in CHOP group was 75.0% (27/36), among which complete remission was seen in 19 patients and partial remission was seen in 8 patients. The therapeutic efficacy was significantly better in combination group than in CHOP group (P<0.05). The 1-year progression-freely and overall survival rates were significantly higher in combination group than in CHOP group (81.2% vs. 52.8%, 93.8% vs. 75.0%, P<0.05). The major adverse events in combination group were infusion-related response which could be well tolerated, and hematological toxicities which were similar to those in CHOP group. CONCLUSIONS: Rituximab increases the therapeutic efficacy of CHOP regimen on newly diagnosed patients with diffuse large B-cell lymphoma, without a clinically significant increase in toxicity. Rituximab plus CHOP can be used as a first-line therapy of diffuse large B-cell lymphoma. PMID- 16351801 TI - [MINE regimen for patients with relapsed or chemo-resistant invasive non Hodgkin's lymphoma]. AB - BACKGROUND & OBJECTIVE: Treatment of relapsed or refractory non-Hodgkin's lymphoma (NHL) remains problematic with no standard salvage chemotherapy regimen. This study was to evaluate the efficacy of MINE (mitoxantrone, mesna/ifosfamide and etoposide) regimen on relapsed or refractory NHL, and observe its toxicity. METHODS: Records of 38 patients with relapsed or refractory invasive NHL, treated with MINE regimen from Jan. 2001 to Jun. 2003, were reviewed. All patients had received at least 1 type of chemotherapy regimen (median, 2 types; range, 1-4 types) with a median of 6 chemotherapy cycles (range, 2-12 cycles). The patients received a median of 4 cycles (range, 2-6 cycles) of MINE regimen. RESULTS: The treatment outcome and adverse events of all patients were assessable. The overall response rate was 47.4%, with a complete response (CR) rate of 15.8%. The response rates were 57.7% in the 26 patients with B-cell lymphoma and 25.0% in the 12 patients with T-cell lymphoma. The 1- and 2-year survival rates were 34.2% and 7.9%, respectively. The major adverse event was myelosuppression: the prevalence of grade III-IV neutropenia was 39.5%, and that of grade III-IV thrombocytopenia was 13.1%. One patient suffered grade III liver toxicity. CONCLUSIONS: MINE regimen was effective for patients with relapsed or refractory invasive NHL, and its toxicity is well tolerated, but the response term is relatively short. Further clinical study on the application of MINE regimen is warranted. PMID- 16351802 TI - [Clinical analysis of liver damage of 116 malignant lymphoma patients with chronic HBV infection after cytotoxic chemotherapy]. AB - BACKGROUND & OBJECTIVE: Chronic hepatitis B virus (HBV) infection increases the prevalence of liver damage and related death of malignant tumor patients. This study was to investigate the prevalence of liver damage and clinical results in lymphoma patients with chronic HBV infection after standard chemotherapy, and assess high risk factors associated with liver damage for better guidance in clinic. METHODS: Records of 116 lymphoma patients with chronic HBV infection, treated with standard chemotherapy from Jan. 1985 to Jan. 2002 in Cancer Center of Sun Yat-sen University, were reviewed to analyze the prevalence of liver damage, clinical results, and related high risk factors. RESULTS: Of the 116 patients, 60 (51.7%) suffered liver damage. According to WHO criteria of liver toxicity, 4 (3.4%) were in grade I, 14 (12.1%) in grade II, 15 (12.9%) in grade III, and 27 (23.3%) in grade IV. After treatment for liver damage, 11 (9.5%) patients completed chemotherapy without delay, 27(23.3%) completed chemotherapy with delay of more than 8 days, 16 (13.8%) terminated chemotherapy, 6 (5.2%) died. Logistic multivariate analysis showed that steroid was a high risk factor of liver damage after chemotherapy. CONCLUSIONS: The prevalence of liver damage is high in lymphoma patients with chronic HBV infection after standard chemotherapy, which led to treatment delay or discontinue, even death. Steroid is a high risk of liver damage. PMID- 16351803 TI - [Correlation of genetic polymorphisms in nucleotide excision repair system to sensitivity of advanced non-small cell lung cancer patients to platinum-based chemotherapy]. AB - BACKGROUND & OBJECTIVE: DNA repair system plays an important role in tumor sensitivity to platinum-based chemotherapy. This study was to examine the correlations of polymorphisms in nucleotide excision repair system to sensitivity of advanced non-small cell lung cancer (NSCLC) to platinum-based chemotherapy. METHODS: Treatment outcomes of 200 advanced NSCLC patients, treated with platinum based chemotherapy, were evaluated. XPC-PAT, XPD Lys751Gln (rs1052559), and ERCC1 C8092A (rs1052559) were genotyped by polymerase chain reaction-amplified fragment length polymorphism (PCR-AFLP) or PCR-restrictive fragment length polymorphism (PCR-RFLP) methods in the 200 patients. Unconditional logistic regression model was used to analyze the correlation of genetic polymorphisms to clinical response. RESULTS: The distributions of XPC-PAT genotypes differed significantly between response group (complete response + partial response) and un-response group (stable disease + progressive disease)(P=0.023). The XPC LL genotype carriers had higher response rate than the SS genotype carriers (OR=3.04; 95% CI=1.25-7.41, P= 0.015). The XPD Lys751Gln and ERCC1 C8092A polymorphisms were not found to be associated with platinum-based chemotherapy. However, these 3 genetic polymorphisms in nucleotide excision repair system had interaction in the drug sensitivity (P=0.021). CONCLUSION: The genetic polymorphisms of XPC-PAT, XPD Lys751Gln, and ERCC1 C8092A in nucleotide excision repair system may be associated with sensitivity of NSCLC patients to platinum-based chemotherapy. PMID- 16351804 TI - [Comparison of treatment efficacy of the second and the third generations of chemoradiotherapy regimens containing platinum on advanced non-small cell lung cancer]. AB - BACKGROUND & OBJECTIVE: Platinum-based chemotherapy regimens are better than other regimens when treating patients with advanced non-small cell lung cancer (NSCLC). The third generation of platinum-based regimens {NP [Navelbine (NVB), cisplatin (DDP)], TP (Taxol, DDP), GP (Gemzar, DDP)} is better than the second generation {MVP [mitomycin (MMC), desacetylvinblastin amide (VDS), DDP], MIP [MMC, ifosfamide (IFO), DDP], EP [etopside (VP-16), DDP], VDS+DDP}. This study was to compare the efficacy between the second and the third generations of platinum-based regimens combined with radiotherapy on advanced non-small cell lung cancer (NSCLC). METHODS: From Jul. 1999 to Dec. 2001, 47 pathologically confirmed advanced NSCLC patients received chemoradiotherapy: 24 received the second generation of platinum-based regimens, 23 received the third generation; all patients received routine external irradiation of (60)Co (65-76 Gy). Characteristics of the patients were comparable. Kaplan-Meier analysis was used to evaluate survival rates, and log-rank test to study differences between the 2 groups. RESULTS: The objective response rates were 41.7% in the second generation group, and 56.5% in the third generation group (Chi (2)=0.53, P=0.47). The median time to progression and median survival time were significantly longer in the third generation group than in the second generation group (12.6 months vs. 6.0 months, Chi(2)=6.93, P=0.01; 14.0 months vs. 9.0 months, Z=-2.17, P=0.03). The 1 , and 2-year survival rates were significantly higher in the third generation group than in the second generation group (56.3% vs. 30.7%, 15.6% vs. 0%, Chi(2)=6.59, P=0.01). The major adverse events were leukocytopenia, nausea and vomiting, radiation-induced esophagitis and pneumonia, which were tolerable. CONCLUSIONS: The third generation of platinum-based regimens in combination with radiotherapy for advanced non-small lung cancer may be more advantageous over the second generation of platinum-based regimens in prolonging survival of advanced NSCLC patients, which would be worth being studied further. PMID- 16351805 TI - [Preliminary report of fludarabine, mitoxantrone and dexamethasone in treating refractory or relapsed multiple myeloma]. AB - BACKGROUND & OBJECTIVE: Multiple myeloma has low complete remission rate and high recurrence rate. Recurrence or relapse of the disease is almost inevitable for most of the patients after several cycles of combined chemotherapy. This study was designed to compare efficacy of fludarabine-based regimen (fludarabine, mitoxantrone and dexamethasone) with that of pirarubicin, vincristine and dexamethasone (VAD) on refractory or relapsed multiple myeloma, and analyze their toxicities. METHODS: Twenty-two patients who had received VAD regimen were taken as a historical control group. A total of 11 patients received FND regimen. The differences between FND group and VAD group were observed and compared. The following indexes were assessed before, during, and after treatment: the partial remission (PR) rate, total response rate, time to achieve PR, the number of patients and time of serum M-component drop to more than 50% of the pre-treatment value, the ratio of myeloma cells in bone marrow drop to less than 5% or more than 80% of pre-treatment level, and the hemoglobin level increase to more than 20 g/Lu the white blood cell and platelet count of the peripheral blood, serum calcium, creatinine, beta2-microglobin and glutamic-oxaloacetic transaminase (GPT) level, adverse events were also analyzed. RESULTS: The PR rate was significantly higher in FND group than in VAD group (45.5% vs. 22.7%, P<0.05). The median time to achieve PR was significantly longer in FND group than in VAD group (76 days vs. 68 days, P<0.05). The occurrence rates of M-component decrease and hemoglobin elevation were significantly higher in FND group than in VAD group [45.5% vs. 22.7%, and 54.5% vs. 18.2%, P<0.05]. There were no significant differences in serum calcium, creatinine and GPT level pre- and post-treatment between the 2 groups. The level of serum beta2-microglobin after treatment was significantly lower than that before treatment in FND group [(1 042.8+/-72.3) microg/L vs. (2 350.2+/-184.0) microg/L, P<0.05]. The nadir white blood cell count was significantly lower in FND group than in VAD group [(0.9+/ 0.46)x10(9)/L vs. (2.09+/-0.6)x10(9)/L, P<0.05], and the occurrence rates of fever and cough was significantly higher in FND group than in VAD group (36.4% vs. 4.5%, 45.5% vs. 9.0%, P<0.05). CONCLUSIONS: Compare with VAD regimen, FND regimen may enhance the PR rate of refractory or relapsed multiple myeloma patients, but it takes longer time to achieve PR, and shows obvious bone marrow inhibition, with no significant renal or hepatic toxicity. FND regimen is effective and safe in treating refractory or relapsed multiple myeloma. PMID- 16351806 TI - [Expression and clinical significance of survivin in bone marrow cells of multiple myeloma patients]. AB - BACKGROUND & OBJECTIVE: Survivin, a member of the inhibitors of apoptosis protein family (IAPs), is overexpressed in many cancers, but not in normal differentiated adult tissues. It is related to poor prognosis and resistance to chemotherapy and radiotherapy in many cancers. This study was designed to detect the expression of Survivin in bone marrow of healthy people, newly diagnosed and relapsed/refractory multiple myeloma (MM) patients, and MM cell line KM3, to explore its correlation to drug resistance and prognosis of MM patients. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect mRNA and protein expression of Survivin in bone marrow samples of 17 newly diagnosed MM patients, 12 relapsed /refractory MM patients, and 9 healthy people, and myeloma cell line KM3. RESULTS: KM3 cells expressed Survivin strongly. The positive rates and the expression levels of Survivin mRNA were significantly lower in healthy people than in newly diagnosed patients and relapsed/refractory patients (22.2 % vs. 70.5 % and 83.3 %, 0.06+/-0.04 vs. 0.31+/-0.14 and 0.69+/-0.24, P<0.01), and were significantly lower in newly diagnosed patients than in relapsed/refractory patients (P<0.05). Survivin protein were not detected in all of the healthy people; while the positive rates of Survivin were 41.4% in newly diagnosed patients and 58.3% in relapsed/refractory patients (P>0.05), and its expression levels was significantly higher in relapsed/refractory patients than in newly diagnosed patients (0.21+/-0.12 vs. 0.11+/-0.07, P<0.05). Among the 7 Survivin-positive newly diagnosed patients, 4 achieved partial remission (PR), 1 achieved MR, and 2 showed no change (NC); among the 10 Survivin-negative newly diagnosed patients, 2 achieved complete remission (CR), 4 achieved PR, 2 achieved MR, and 2 showed NC. The effective (CR+PR+MR) rate was slightly higher in Survivin-negative patients than in Survivin-positive patients (80.0% vs. 71.4%). CONCLUSION: High expression of Survivin may be relates to drug resistance in some patients. Since Survivin only expressed in bone marrow cells of multiple myeloma, but not in healthy people, it can be a new target to reverse drug resistance in myeloma cells. PMID- 16351807 TI - [Clinical analysis of 67 patients with brain metastases from breast cancer]. AB - BACKGROUND & OBJECTIVE: The incidence of brain metastases from breast cancer seems to be increasing with the improvement of systemic control and survival prolongation. This study was to analyze the clinical characteristics, treatment modalities, and prognostic factors of brain metastases from breast cancer. METHODS: The time from primary diagnosis of breast cancer to brain metastases of 67 breast cancer patients were 0-15 years, and the median time was 2.5 years. Of the 67 patients, 3 received surgery plus radiotherapy, 30 received radiotherapy alone, 3 received chemotherapy alone, 26 received radiochemotherapy, and 5 received no treatment. Kaplan-Meier analysis and log-rank test were used to analyze survival status. RESULTS: Headache was the most common symptom of brain metastases from breast cancer. The median survival time was 4 years. After brain metastases, the median survival time was 11 months. Menstrual status, number of brain metastases, systemic metastases, and different treatment modalities had no correlations to survival time of the patients. CONCLUSIONS: The prognosis of brain metastases from breast cancer is still poor. Radiotherapy remains the standard treatment of brain metastases. The function of chemotherapy needs further study. PMID- 16351808 TI - [A randomized controlled trail of taxol-based combination regimens for advanced gastric cancer]. AB - BACKGROUND & OBJECTIVE: Standard chemotherapy for advanced gastric cancer remains undefined. Phase II trials show that taxol is effective in treating advanced gastric cancer. This multi-center prospective open randomized controlled study was to compare the efficacy of Taxol plus calcium folinate (CF)/5-fluorouracil (5 FU), Taxol plus oxaliplatin (OXA), and CF/5-FU plus cisplatin (DDP) on advanced gastric cancer, and analyze their toxicities. METHODS: Patients with measurable unresectable and/or metastatic gastric carcinoma were randomized into CF/5-FU+DDP (control), CF/5-FU + Taxol, and Taxol + OXA groups, and received up to 8 cycles of chemotherapy. Treatment efficacy and adverse events were evaluated according to WHO criteria. RESULTS: A total of 180 patients were enrolled from May 2002 to May 2004, and randomized into the 3 groups; each group contained 60 patients. Of the 180 patients, 14 received 2 cycles of chemotherapy, 49 received 4 cycles, and 103 received 8 cycles. Treatment outcomes of 166 cases were evaluable. The response rate (RR) of naive patients or the patients with retroperitoneal lymph node metastasis was significantly higher in CF/5-FU+Taxol and Taxol+OXA groups than in control group (50.00% and 80.00% vs. 20.75%, P<0.05; 65.96% and 85.71% vs. 36.36%, P<0.05). But the RR of the patients with liver metastasis was similar among the 3 groups (28.57% and 39.13% vs. 34.62%, P>0.05). The occurrence rates of nausea/vomiting, anepithymia, stomatitis, and kidney damage were lower in study groups than in control group, but the occurrence rates of myelosuppression and peripheral nerve damage were higher in study groups than in control group. Allergic response occurred in 7 (5.88%) patients in study group, and 3 (2.52%) of them were serious. There was no treatment-related death. CONCLUSIONS: Despite its hematotoxicity, the treatment efficacy of Taxol-based combination regimens on advanced gastric cancer is better than that of CF/5-FU + DDP regimen with tolerable toxicities. We recommend Taxol-based combination regimens as first-line regimens for advanced gastric cancer. PMID- 16351809 TI - [Impact of preoperative lymphatic chemotherapy on relapse and metastasis of breast cancer and its mechanism]. AB - BACKGROUND & OBJECTIVE: Although it is reported that lymphatic chemotherapy could raise drug concentrations in local lymph nodes and prolong survival time of patients with gastrointestinal tumors, its effect on breast cancer has not been explored. This study was to explore the impact of lymphatic chemotherapy on relapse and metastasis of breast cancer, and to investigate the mechanism. METHODS: Sixty patients with breast cancer of stage II-III were randomized into 2 groups: 40 patients in Epi-CH (carbon activated absorbing epirubicin) group were injected with 10 mg of Epi-CH in the tissue around primary tumor 72 h before modified radical resection; 20 patients in control group were injected with 10 mg of aqueous epirubicin in the same region. The stained nodes full of tumor cells in Epi-CH group and non-stained nodes in control group were selected. The apoptotic index (AI) of cancer cells in metastatic axillary lymph node was calculated by TUNEL method; the expression of Fas/Fas-L proteins was examined by SP immunohistochemistry; the relapse and metastatic rate was compared. RESULTS: The AI of cancer cells in metastatic axillary lymph node was significantly higher in Epi-CH group than in control group [(9.5+/-2.7)% vs. (3.8+/-1.4)%, P<0.01]. The expression of Fas protein was significantly higher in Epi-CH group than in control group (P<0.05), but the expression of Fas-L protein had no difference between the 2 groups (P>0.05). No chemotherapy-related local and whole body reaction occurred in both groups. The relapse and metastatic rate was significantly lower in Epi-CH group than in control group (P<0.05). CONCLUSION: Preoperative Epi-CH lymphatic chemotherapy could suppress relapse and metastasis of breast cancer, which might through up-regulating expression of Fas protein and inducing apoptosis of axillary metastasis cells. PMID- 16351810 TI - [Predictive value of P53 expression in selecting first-line chemotherapy regimen for advanced epithelial ovarian carcinoma]. AB - BACKGROUND & OBJECTIVE: The first-line adjuvant chemotherapy regimens of ovarian cancer mainly include TC (paclitaxel combined with carboplatin) and PC (cisplatin combined with cyclophosphamide) protocols. This study was to investigate the correlation of P53 expression to treatment outcome of ovarian cancer patients received the above 2 protocols, and explore the predictive value of P53 expression in selecting chemotherapy regimen. METHODS: Records of 53 patients with advanced epithelial ovarian cancer (stage IIIc), treated with TC or PC regimen, were analyzed retrospectively. The expression of P53 was detected by immunohistochemistry. The complete response (CR) rate and progression-free survival (PFS) were compared between TC and PC groups according to P53 status. RESULTS: Of the 53 patients, 22 were P53 positive, of which 13 received TC regimen and 9 received PC regimen; the CR rate was slightly higher in TC group than in PC group (76.9% vs. 33.3%, P=0.054), and PFS was significantly longer in TC group than in PC group (102 weeks vs. 43 weeks, P=0.040). In the subgroup of P53-negative patients, TC group had similar CR rate and PFS to PC group. Multivariate analysis showed that the size of residue was an independent prognostic factor. CONCLUSIONS: P53 detection may play a role in selecting first line chemotherapy for advanced epithelial ovarian cancer patients: TC regimen is preference for P53-positive patients, PC regimen may be a choice for P53-negative patients. These recommendations should be tested in further trails with large samples. PMID- 16351811 TI - [Progression of adjuvant chemotherapy for colon cancer]. AB - Postoperative adjuvant chemotherapy is benefit to survival of stage III colon cancer patients. Regimens containing 5-fluorouracil (5-FU) are used as the main therapy, including 5-FU/Lev, Mayo (FU/LV), protracted venous infusion of 5-FU, orally administered fluorouracil (UFT, Xeloda), and so on. 5-FU combined with oxaliplatin may improve therapeutic efficacy; however, further study on therapeutic efficacy of 5-FU combined with irinotecan is needed. The survival benefit of postoperative adjuvant chemotherapy for stage II colon cancer is controversial now; while postoperative adjuvant chemotherapy for high-risk colon cancer patients should be considered. The progression of postoperative adjuvant chemotherapy for colon cancer patients was reviewed in the article. PMID- 16351812 TI - Editorial - relaunching ZOOLOGY. PMID- 16351813 TI - Membrane-permeabilizing polypeptides of amoebae - constituents of an archaic antimicrobial system. AB - Amoebae may be viewed as primitive, actively phagocytosing eukaryotic cells, many of which use bacteria as a major nutrient source. At a very archaic level, amoebae exert mechanisms which kill bacteria comparable to those found in phagocytic cells of higher organisms. Accordingly, it is tempting to suggest that the ancestors of effector cells of the innate immune system were bacteria-feeding amoebae and that their molecular armament is ancient. Here, we summarize the characteristics of antimicrobial and cytolytic 77-residue polypeptides from the protozoon Entamoeba histolytica for which correlates were found in effector cells of the mammalian immune systems. Based on the current knowledge about these small membrane-destabilizing proteins of phylogenetically extremely diverse origin, we discuss similarities and differences in their structure and activities. PMID- 16351814 TI - Evidence for perforin-like activity associated with earthworm leukocytes. AB - Earthworm (Eisenia fetida) coelomic fluid contains several leukocytes (coelomocytes): basophils, acidophils and neutrophils as well as chloragocytes. Small coelomocytes and coelomocyte lysate are cytotoxic for the tumor cell target K562. The expression of a lytic factor was investigated by immunocytochemistry using light and transmission electron microscopy. A rat-anti-mouse-perforin-mAb labeled mainly small coelomocytes (nearly 20%) as visualized by light microscopy. TEM analysis using immunogold showed a homogenous labeling in the cytoplasm of small coelomocytes. The highest number of immunogold particles was estimated in coelomocytes with many small cytoplasmic granules. Coelomocytes with large lysosomal granules were also labeled but less intensely. No antibody binding was observed for chloragocytes either in light or electron microscopy. This suggests that the perforin-like activity is associated with only one cell type and that chloragocytes are responsible for other lytic activities. MALDI-MS revealed calreticulin usually associated with perforin in mammalian cells that mediate lysis (e.g. NK, CTL). Together, results strongly suggest the presence of putative perforin in earthworms. This in turn supports the hypothesis that perforin is a conserved component important in immune defense during evolution. PMID- 16351815 TI - Role of epithelial cells and programmed cell death in Hydra spermatogenesis. AB - Spermatogenesis in higher animals is a tightly regulated process, in which survival and death of sperm precursor cells depends on the presence of somatic cells in gonads. In the basal metazoan Hydra spermatogenesis takes place in anatomically simple testes and in the absence of accessory structures. Hydra sperm precursors are derived from interstitial stem cells. Here we show that large numbers of sperm precursors in testes of Hydra vulgaris undergo programmed cell death (apoptosis) and that ectodermal epithelial cells phagocytose the apoptotic sperm precursors. This is surprising since so far no evidence has been reported that epithelial cells are directly involved in germ cell differentiation in Hydra. We propose that, similar to Sertoli cells in mammals, in Hydra epithelial cells support and perhaps even control spermatogenesis. PMID- 16351816 TI - Physiological ecology of Mediterranean Blue Tits (Parus caeruleus). I. A test for inter-population differences in resting metabolic rate and thermal conductance as a response to hot climates. AB - Blue Tits (Parus caeruleus) are widely distributed throughout Europe, reaching the southern limit of their range on islands in the Mediterranean and in northern Africa. On Corsica, one population located at Pirio in the Fango Valley breeds roughly one month later than populations in adjacent valleys or on the continent, thus exposing nestlings to high ambient temperatures (T(a)). We tested the hypothesis that nestlings and possibly adult Blue Tits at Pirio would exhibit a reduction in resting metabolic rate (MR) and an increase in thermal conductance as a physiological response to high T(a). We compared the thermoregulatory response and evaporative water loss for nestlings from Pirio in Corsica and one continental site (Vic-le-Fesq) and for adults from two Corsican (Pirio and Muro) and one continental site (La Rouviere). At 12-14 days of age, nestlings from Pirio showed two distinct thermoregulatory patterns. Nestlings under 8.0 g behaved as heterotherms, whereby MR was correlated only with body temperature. At body masses above 8.0 g nestlings progressively acquired the ability to regulate T(b) and at masses >9.0 g they behaved as homeotherms. When considering homeothermic nestlings and adults, population of origin did not affect either thermal conductance or resting MR. For homeothermic nestlings, mass-specific resting MR (mW x g-(1)) was 15.5 +/- 2.6 and 17.5 +/- 2.5 for nestlings from Vic le-Fesq and Pirio, respectively. For adults, mass-specific resting MR (mW x g (1)) was 17.5 +/- 2.0, 17.8 +/- 1.6, and 17.9 +/- 1.0 for birds from Pirio, Muro, and La Rouviere, respectively. Although there was a weak but positive effect of T(a) on evaporative water loss for homeothermic nestlings, no such trend was evident for adults over the range of T(a) tested in this study. We thus find no evidence to indicate that either nestlings or adults exhibit the exponential increase in evaporative water loss associated with the non-convective regulation of T(b) within the range of T(a) tested (roughly P. cylindrica >P. lobata >P. australiensis>P. lutea. Out of the 111 interspecific encounters studied, in only 5.4% reciprocal interactions were recorded, and in a single case, the opposite directionality of hierarchy was documented. Allogeneic encounters were also observed. A single major effector mechanism, an overgrowth (together with secondary outcomes such as the formation of small points of rejection, bleaching and pink color formation along a narrow peripheral belt of contacting tissues), was the only response in all 10 xenogeneic and 5 allogeneic combinations. In some massive colonies, a long contacting line of up to 50 cm was established. No sign for allelopathy, stand off or rejection from a distance (i.e., by sweeper tentacles, sweeper polyps) was observed. Results are discussed with the accumulated data on Porites species from different reefs, worldwide, confirming that this genus is commonly lower in the hierarchy of xenogeneic interactions. PMID- 16351823 TI - Brachyury homolog (HpTa) is involved in the formation of archenteron and secondary mesenchyme cell differentiation in the sea urchin embryo. AB - Sea urchin Brachyury homolog (HpTa) is expressed exclusively in the vegetal plate and secondary mesenchyme cells in the embryos of sea urchin Hemicentrotus pulcherrimus. In order to gain insights into the role of HpTa during sea urchin development, we designed experiments to perturb the embryo by inducing ectopic overexpression of HpTa by injecting fertilized eggs with HpTa mRNA. The overexpression of HpTa resulted in suppression of the formation of vegetal plate and secondary mesenchyme cells. We assume that the interaction of HpTa with unknown factors is required for the activation of the HpTa target genes, and that the excess amount of HpTa proteins produced from injected HpTa mRNA depletes the co-factors. In consequence, the target genes of HpTa would be repressed by the overexpression of HpTa. We suggest that HpTa is involved in the formation of the vegetal plate and the differentiation of secondary mesenchyme cells. PMID- 16351824 TI - Perspectives on cardiac physiological ontogeny in crustaceans. AB - Crustacean embryonic and larval systems offer a unique and valuable tool for furthering our understanding of both developmental processes and physiological regulatory mechanisms. The diverse array of developmental patterns exhibited by crustaceans allows species choice to be based on the specific questions being investigated, where defined larval forms are chosen based on their developmental pattern, degree of maturation or regulatory capabilities. However, this great diversity in developmental patterns, as well as crustacean diversity, can also confound ones ability to define or identify species for investigation. These issues are addressed and suggestions put forth to clarify some of the problems. The complexity and overlapping nature of adult cardio-regulatory systems makes teasing them apart difficult. Embryonic and larval systems exhibit varying degrees of regulatory complexity depending on developmental stage and ontogenetic pattern. This can allow complex adult regulatory systems to be teased apart temporally, as the developing animal builds regulatory pathways. Equally important is the nature of crustacean larvae; many undergo dramatic metamorphoses in cases where the larvae have adaptations to environments different to those of the adult. During environmental transitions physiological adaptations to immediate change should take precedence over long-term adult adaptations. It is therefore possible to look at physiological responses as a function of developmental/environmental adaptation, independent of adult functions. PMID- 16351825 TI - A comparative study of embryonic development of Japanese quail selected for different patterns of postnatal growth. AB - Patterns of early embryonic development have traditionally been viewed as invariant within vertebrate taxa. It has been argued that the specific differences which are found arise during the later stages of development. These differences may be a result of allometry, heterochrony or changes in relative growth rates. To test whether early embryonic development is indeed invariant, or whether selection of adult characteristics can alter embryonic growth, we compared embryonic development in birds selected for different patterns of postnatal growth. Using quail lines selected for high and low body mass, we compared somite formation, and muscle and feather development. We obtained data that showed changes in the rate of myotome formation in the brachial somites which contribute to muscle formation in the limbs and thorax. We think these observations are connected with intraspecific changes in adult morphology, ie., breast muscle size. Our findings suggest that selection for late ontogenetic/adult stages affects early embryonic development. PMID- 16351826 TI - Wing folding and the functional morphology of the wing base in Coleoptera. AB - The wing unfolding of Pachnoda marginata was examined using digital video (50 half-fps) and high speed video sequences (1000 fps), and the skeleto-muscular apparatus of the metathorax was described. Left and right hind wing are able to promote independently of each other. The hind wings do not unfold instantly when the elytra are lifted and may also reach the flight position (and beat) while still folded. Wing promotion is exhaustible and the time needed for unfolding varies considerably. These observations strongly suggest a muscular control. Wing unfolding is probably triggered by contraction of M. pleura alaris and a resulting proximad movement of the 3rd axillary sclerite, pulling the Media posterior backwards, while the Radius anterior is held by the basalar muscle as the antagonist. Our findings are in clear contrast to the earlier assumption that the hind wings of Coleoptera either unfold or fold due to intrinsic elasticity. The specific wing folding and unfolding mechanisms are autapomorphic character states of Coleoptera. They were maintained during evolution even though considerable variations of skeletal thoracic structures, musculature and venation occurred. (Additional material is available from the Zoology web page: http://www.urbanfischer.de/journals/zoology). PMID- 16351827 TI - Buccal oscillation and lung ventilation in a semi-aquatic turtle, Platysternon megacephalum. AB - Movements of the hyobranchial apparatus in reptiles and amphibians contribute to many behaviors including feeding, lung ventilation, buccopharyngeal respiration, thermoregulation, olfaction, defense and display. In a semi-aquatic turtle, Platysternon megacephalum, x-ray video and airflow measurements from blowhole pneumotachography show no evidence that above water hyobranchial movements contribute to lung inflation, as in the buccal or gular pump of amphibians and some lizards. Instead, hyobranchial movements produce symmetrical oscillations of air into and out of the buccal cavity. The mean tidal volume of these buccal oscillations is 7.8 times smaller than the mean tidal volume of lung ventilation (combined mean for four individuals). Airflow associated with buccal oscillation occurs in the sequence of inhalation followed by exhalation, distinguishing it from lung ventilation which occurs as exhalation followed by inhalation. No fixed temporal relationship between buccal oscillation and lung ventilation was observed. Periods of ventilation often occur without buccal oscillation and buccal oscillation sometimes occurs without lung ventilation. When the two behaviors occur together, the onset of lung ventilation often interrupts buccal oscillation. The initiation of lung ventilation was found to occur in all phases of the buccal oscillation cycle, suggesting that the neural control mechanisms of the two behaviors are not coupled. The pattern of occurrence of both buccal oscillation and lung ventilation was found to vary over time with no obvious effect of activity levels. PMID- 16351828 TI - Brainstem lateral line responses to sinusoidal wave stimuli in the goldfish, Carassius auratus. AB - Extracellular recordings were made from single lateral line units in the medial octavolateralis nucleus in the brainstem of goldfish, Carassius auratus. Units were defined as receiving lateral line input if they responded to the water motions generated by a stationary, sinusoidally oscillating sphere and/or a moving sphere but not to airborne sound and vibrations. Units which responded to airborne sound or vibrations were assumed to receive input from the inner ear and were not further investigated. Responses of lateral line units were quantified in terms of the number of evoked spikes and the degree of phase-locking to a 50 Hz vibrating sphere presented at various stationary locations along the side of the fish. Receptive fields were characterized based on spike rate, degree of phase locking and average phase angle as a function of sphere location. Four groups of units were distinguished: 1, units with receptive fields comparable to those of primary afferents; 2, units with receptive fields which consisted of one excitatory and one inhibitory area; 3, units with receptive fields which consisted of more than two excitatory and/or inhibitory areas; 4, units with receptive fields which consisted of a single excitatory or a single inhibitory area. The receptive fields of most units were characterized by adjacent excitatory and inhibitory areas. This organization is reminiscent of excitatory inhibitory receptive field organizations in other vertebrate sensory systems. PMID- 16351829 TI - Zoology www guide. AB - As in the previous issue of Zoology, we introduce here web sites which are of interest for zoologists. Readers are welcome to make website suggestions for future editions of Zoology and should contact tbosch@zoologie.uni-kiel.de. In this issue the focus is on amphibians and reptiles. PMID- 16351831 TI - The genetic control of eye development and its implications for the evolution of the various eye-types. PMID- 16351832 TI - Private and collective interests; conflicts and solutions: the central theme of current thinking in evolutionary biology. AB - The statement made by the population geneticist Theodosius Dobzhansky (1973): "Nothing in Biology Makes Sense Except in the Light of Evolution", is often quoted as a crucially important generalization on the nature of biology. I am inclined to consider as equally important the statement: "Nothing in Evolutionary Biology Makes Sense Except in the Light of Conflicts between Parts and Systems." This generalization takes account of the dynamic nature of biological phenomena, but also of the fact that the study of transitions from autonomous units to cooperative systems has become one of the most exciting and scientifically rewarding enterprises in all of organismic biology. The problems encountered and the speculations generated in the course of this enterprise will be either of the more unit-centered or of the more system-centered type, most biologists tending to lean towards one or the other. This explains why evolutionary biology is fraught with so many antagonistic attitudes and polarizing points of view. In this essay I want specifically to draw attention to and discuss the following issues which in recent years have polarized biologists: the dual nature of genes; the logic of Hamilton's rule; the relationship between kin selection, signalling networks and systemic manipulation; the semantic problem of progress in evolution; and the evolutionary consequences of the vastly differing time scales over which genotypic and phenotypic information processing occurs in higher animals. PMID- 16351833 TI - Mammalian gene expression in hypoxic conditions. AB - Hypoxia induces gene expression of specific genes such as erythropoietin (Epo) and vascular endothelial growth factor (VEGF) that allow physiological adaptation to the environmental conditions at the cellular, local, and systemic levels. Reduced oxygenation is also a common precursor of many pathological processes, including coronary artery defects, ischemia, and malignant tumour formation. The hypoxia-inducible transcription factor HIF-1, a heterodimer consisting of the oxygen-regulated alpha-subunit and the constitutively expressed beta or ARNT subunit, serves as a master regulator of oxygen-dependent gene expression. We observed that upon hypoxic exposure of HeLa cells in tonometer, accumulation of HIF-1alpha occurred within two minutes, while reoxygenation strongly reduced HIF 1alpha levels within four to eight minutes. Thus, hypoxia leads to a rapid cellular adaptation. In another line of investigation, we analysed the impact of hypoxia-independent overexpression of Epo in transgenic mice. Despite a hematocrit of about 80% the transgenic mice did not develop hypertension or thromboembolic complications. PMID- 16351834 TI - Cellular osmoregulation: beyond ion transport and cell volume. AB - All cells are characterized by the expression of osmoregulatory mechanisms, although the degree of this expression is highly variable in different cell types even within a single organism. Cellular osmoregulatory mechanisms constitute a conserved set of adaptations that offset antagonistic effects of altered extracellular osmolality/environmental salinity on cell integrity and function. Cellular osmoregulation includes the regulation of cell volume and ion transport but it does not stop there. We know that organic osmolyte concentration, protein structure, cell turnover, and other cellular parameters are osmoregulated as well. In this brief review two important aspects of cellular osmoregulation are emphasized: 1) maintenance of genomic integrity, and 2) the central role of protein phosphorylation. Novel insight into these two aspects of cellular osmoregulation is illustrated based on two cell models, mammalian kidney inner medullary cells and teleost gill epithelial cells. Both cell types are highly hypertonicity stress-resistant and, therefore, well suited for the investigation of osmoregulatory mechanisms. Damage to the genome is discussed as a newly discovered aspect of hypertonic threat to cells and recent insights on how mammalian kidney cells deal with such threat are presented. Furthermore, the importance of protein phosphorylation as a core mechanism of osmosensory signal transduction is emphasized. In this regard, the potential roles of the 14-3-3 family of phospho-protein adaptor molecules for cellular osmoregulation are highlighted primarily based on work with fish gill epithelial cells. These examples were chosen for the reader to appreciate the numerous and highly specific interactions between stressor-specific and non-specific pathways that form an extensive cellular signaling network giving rise to adaptive compensation of hypertonicity. Furthermore, the example of 14-3-3 proteins illustrates that a single protein may participate in several pathways that are non-specific with regard to the type of stress and, at the same time, in stress-specific pathways to promote cell integrity and function during hypertonicity. PMID- 16351835 TI - Coping with excess salt: adaptive functions of extrarenal osmoregulatory organs in vertebrates. AB - In all organisms, changing environmental conditions require appropriate regulatory measures to physiologically adjust to the altered situation. Uptake of excess salt in non-mammalian vertebrates having limited or no access to freshwater is balanced by extrarenal salt excretion through specialized structures called 'salt glands'. Nasal salt glands of marine birds are usually fully developed in very early stages of their lives since individuals of these species are exposed to salt soon after hatching. In individuals of other bird species, salt uptake may occur infrequently. In these animals, glands are usually quiescent and glandular cells are kept in a fairly undifferentiated state. This is the situation in 'naive' ducklings, Anas platyrhynchos, which have never been exposed to excess salt. When these animals become initially osmotically stressed, the nasal glands start to secrete a moderately hypertonic sodium chloride solution but secretory performance is meager. Within 48 h after the initial stimulus, however, the number of cells per gland is elevated by a factor of 2-3, the secretory cells differentiate and acquire full secretory capacity. During this differentiation process, extensive surface specializations are formed. The number of mitochondria is increased and metabolic enzymes and transporters are upregulated. These adaptive growth and differentiation processes result in a much higher efficiency of salt excretion in acclimated ducklings compared with naive animals. Receptors and signal transduction pathways in salt gland cells controling the adaptive processes seem to be the same as those controling salt secretion, namely muscarinic acetylcholine receptors and receptors for vasoactive intestinal peptide. This review focusses on signal transduction pathways activated by muscarinic receptors which seem to fine-tune salt secretion in salt adapted ducklings and may control adaptive growth and differentiation processes in the nasal gland of naive animals. PMID- 16351836 TI - Cytoskeletal mechanisms of neuronal morphogenesis. AB - The cytoskeleton is the major intracellular structure that determines the morphology of a neuron. Thus, mechanisms that ensure a precisely regulated assembly of cytoskeletal elements in time and space have an important role in the development from a morphologically simple neuronal precursor cell to a complex polarized neuron that can establish contacts to several hundreds of other cells. Here, cytoskeletal mechanisms that underlie the formation of neurites, directed elongation and stabilization of neuronal processes are summarized. It has become evident that different cytoskeletal elements are highly crosslinked with each other by several classes of specific linker proteins. Of these, microtubule associated proteins (MAPs) appear to have an important role in connecting the microtubule skeleton to other cytoskeletal filaments and plasma membrane components during neuronal morphogenesis. Future experiments will have to elucidate the function and the regulation of the neuronal cytoskeleton in an authentic nervous system environment during development. Recent approaches are discussed at the end of this article. PMID- 16351837 TI - Axon guidance processes in the retinotectal and vomeronasal projection are controlled by Eph receptor tyrosine kinases and ephrins. AB - The Eph family of receptor tyrosine kinases and their 'ligands', the ephrins, have been shown to play key roles in a number of different developmental processes such as cell migration, boundary formation, axon guidance, synapse formation and vasculogenesis. Here, we summarize recent findings derived from investigating the role of the EphA family during development of the retinotectal and vomeronasal projection uncovering a role of ephrin-A molecules as axon guidance receptors. PMID- 16351838 TI - Cellular processes in the amygdala: gates to emotional memory? AB - The amygdala is considered a core structure of the so-called limbic system and has been implicated in a variety of functions, including emotional interpretation of sensory information, emotional arousal, emotional memory, fear and anxiety, and related clinical disorders. Despite the clinical and functional importance of the amygdala, it is only recently that some general principles of intra amygdaloid mechanisms of signal processing that are relevant for fear behavior and memory have emerged from behavioral, anatomical, electrophysiological, and neurochemical studies performed in the amygdala of various mammalian species in vivo, in situ and in vitro. PMID- 16351839 TI - Functional implications of temporal structure in primate cortical information processing. AB - Processing of information in the cerebral cortex of primates is characterized by distributed representations and processing in neuronal assemblies rather than by detector neurons, cardinal cells or command neurons. Responses of individual neurons in sensory cortical areas contain limited and ambiguous information on common features of the natural environment which is disambiguated by comparison with the responses of other, related neurons. Distributed representations are also capable to represent the enormous complexity and variability of the natural environment by the large number of possible combinations of neurons that can engage in the representation of a stimulus or other content. A critical problem of distributed representation and processing is the superposition of several assemblies activated at the same time since interpretation and processing of a population code requires that the responses related to a single representation can be identified and distinguished from other, related activity. A possible mechanism which tags related responses is the synchronization of neuronal responses of the same assembly with a precision in the millisecond range. This mechanism also supports the separate processing of distributed activity and dynamic assembly formation. Experimental evidence from electrophysiological investigations of non-human primates and human subjects shows that synchronous activity can be found in visual, auditory and motor areas of the cortex. Simultaneous recordings of neurons in the visual cortex indicate that individual neurons synchronize their activity with each other, if they respond to the same stimulus but not if they are part of different assemblies representing different contents. Furthermore, evidence for synchronous activity related to perception, expectation, memory, and attention has been observed. PMID- 16351840 TI - Complex functions of the brain. AB - Over the course of the last 50 years it has been possible to solve a number of basic problems in neurobiology. Interest is now turning more and more to problems concerning so-called "higher" brain functions, including cognition. Examples from the visual system in primates are presented. First relatively elementary problems are illustrated, such as how long it takes to perceive an object or to respond to a stimulus or combinations of stimuli. Top-down modification of perception by expectation is demonstrated in an illusion of misdirected gaze. Interdisciplinary questions straddling the sciences and the humanities are also approached, such as which part of the brain mediates conscious perception. Finally, the problem of causality and freedom of will is addressed, taking into account the knowledge accumulated in the neurosciences during the last 5 decades. PMID- 16351841 TI - Assessing non-metric skeleton characters as a morphological tool. AB - The appearance of non-metric skeletal characters in vertebrates results mainly from basic genetic control, as proved and documented for house mice and humans. Although the heritability of non-metric traits, mainly the presence of foramina and similar structures for blood vessels and nerves or dental occlusal variants, have been evaluated as rather low, the simultaneous consideration of several traits allows estimation of epigenetic variation in time and space as a result of genetic relationship. Thus, the main use of non-metric characters has been aimed at assessing epigenetic variability and divergence among populations. Applications extend from the problem of genetic isolation of populations, the lack of reproductive contact, detection of genetic drift, systematic studies to clarify species taxonomy, to phylogenetic interpretation. Additionally, non directional deviations from bilateral symmetry in non-metric characters, e.g. fluctuating asymmetry, could be caused by current environmental conditions in general. Fluctuating asymmetry is regarded as a measure of developmental instability to indicate the presence of genomic changes or the influence of contamination and/or habitat deterioration, and is also used as a further population parameter with integrated information. However, standpoints on the use of fluctuating asymmetry are quite inconsistent, fluctuating themselves between considering it a powerful biomonitoring tool to being merely a curious scientific toy. PMID- 16351842 TI - Comparative methods in developmental biology. AB - The need for a phylogenetic framework is becoming appreciated in many areas of biology. Such a framework has found limited use in developmental studies. Our current research program is therefore directed to applying comparative and phylogenetic methods to developmental data. In this paper, we examine the concepts underlying this work, discuss potential difficulties, and identify some solutions. While developmental biologists frequently make cross-species comparisons, they usually adopt a phenetic approach, whereby degrees of overall similarity in development are sought. Little emphasis is placed on reconstructing the evolutionary divergence in developmental characters. Indeed, developmental biologists have historically concentrated on apparently 'conserved' or 'universal' developmental mechanisms. Thus, there has been little need for phylogenetic methodologies which analyse specialised features shared only within a subset of species (i.e., synapomorphies). We discuss the potential value of such methodologies, and argue that difficulties in adapting them to developmental studies fall into three interlinked areas: One concerns the nature and definition of developmental characters. Another is the difficulty of identifying equivalent developmental stages in different species. Finally the phylogenetic non independence of developmental characters presents real problems under some protocols. These problems are not resolved. However, it is clear that the application of phylogenetic methodology to developmental data is both necessary and fundamental to research into the relationship between evolution and development. PMID- 16351843 TI - Zoology meets Botany: establishing intracellular organelles by endosymbiosis. AB - One of the most citated characteristics of eukaryotic cells are mitochondria and in the case of phototrophic cells, the plastids. These organelles are of eubacterial origin and contain a remnant genome. Here, we present hypotheses concerning the origin of the first mitochondrium-harboring cell and show the evolution of primary, secondary and tertiary plastids. Furthermore we discuss models explaining why plastids have to maintain their own genome. PMID- 16351844 TI - Hydrogenosomes: convergent adaptations of mitochondria to anaerobic environments. AB - Hydrogenosomes are membrane-bound organelles that compartmentalise the final steps of energy metabolism in a number of anaerobic eukaryotes. They produce hydrogen and ATP. Here we will review the data, which are relevant for the questions: how did the hydrogenosomes originate, and what was their ancestor? Notably, there is strong evidence that hydrogenosomes evolved several times as adaptations to anaerobic environments. Most likely, hydrogenosomes and mitochondria share a common ancestor, but an unequivocal proof for this hypothesis is difficult because hydrogenosomes lack an organelle genome - with one remarkable exception (Nyctotherus ovalis). In particular, the diversity of extant hydrogenosomes hampers a straightforward analysis of their origins. Nevertheless, it is conceivable to postulate that the common ancestor of mitochondria and hydrogenosomes was a facultative anaerobic organelle that participated in the early radiation of unicellular eukaryotes. Consequently, it is reasonable to assume that both, hydrogenosomes and mitochondria are evolutionary adaptations to anaerobic or aerobic environments, respectively. PMID- 16351845 TI - Mollusc/algal chloroplast symbiosis: how can isolated chloroplasts continue to function for months in the cytosol of a sea slug in the absence of an algal nucleus? AB - A marine sea slug, Elysia chlorotica, has acquired the ability to carry out photosynthesis as a result of forming an intracellular symbiotic association with chloroplasts of the chromophytic alga, Vaucheria litorea. The symbiont chloroplasts (kleptoplasts) are functional, i.e. they evolve oxygen and fix CO(2) and actively transcribe and translate proteins for several months in the sea slug cytosol. Considering the dependency of plastid function on nuclear genes, the level of kleptoplast activity observed in the animal cell is quite remarkable. Possible factors contributing to this long-lasting functional association that are considered here include: the presence of an algal nuclear genome in the sea slug, autonomous chloroplasts, unusual chloroplast/protein stability, re directing of animal proteins to the kleptoplast, and lateral gene transfer. Based on our current understanding, the acquisition and incorporation of intact algal plastids by E. chlorotica is aided by the robustness of the plastids and the long term functional activity of the kleptoplasts appears to be supported by both plastid and protein stability and contributions from the sea slug. PMID- 16351846 TI - Seasonal adaptations and the role of lipids in oceanic zooplankton. AB - Oceanic zooplankton species exhibit quite diverse life history traits. A major driving force determining their life strategies is the seasonal variability in food supply, which is most pronounced in polar oceans where fluctuations in primary production are extreme. Seasonal adaptations are closely related to the trophic level of zooplankters, with strongest pressures occurring on herbivorous organisms. The dominant grazers, calanoid copepods and krill (Euphausiacea), have developed fascinating solutions for successful overwintering at higher latitudes. They usually exhibit a very efficient storage and utilization of energy reserves to reduce the effect of a highly seasonal primary production. The predominant larger Calanus species from the Arctic and Calanoides acutus from the Antarctic biosynthesize large amounts of high-energy wax esters with long-chain monounsaturated fatty acids and alcohols (20:1 and 22:1 isomers) as major components. They survive the dark season at depth in a stage of dormancy called diapause. In contrast, the Antarctic Calanus propinquus, a winter-active species, synthesizes primarily triacylglycerols, which are dominated by long-chain monounsaturated fatty acids with 22 carbon atoms (2 isomers) and yield even higher calorific contents. The omnivorous and carnivorous species, which are less subjected to seasonal food shortage, usually do not exhibit such an elaborate lipid biosynthesis. Herbivores usually do not utilize much of their enormous lipid reserves for overwintering, but channel this energy towards reproductive processes in late winter/early spring. Timing of reproduction is critical especially at high latitudes due to the short production period, and lipid reserves ensure early spawning independent of external resources. These energetic adaptations (dormancy, lipid storage) are supplemented by other life strategies such as extensive vertical migrations, change in the mode of life, and trophic flexibility. PMID- 16351847 TI - Finding food in the open ocean: foraging strategies in Humboldt penguins. AB - Penguins are excellent "model" organisms allowing us to study the behaviour of marine homeotherms at sea. Penguins regularly return to their breeding colonies, enabling biologists to equip them with remote sensing devices such as physiological or behavioural data-loggers, radio- or satellite transmitters. Foraging trips at sea can last from days to weeks and after return of the birds to their breeding sites, the devices can easily be removed for analysis of on board stored data, yielding a wealth of information. Investigation of penguin behaviour at sea becomes particularly revealing when other sources of information can be matched to the data set, such as satellite data on wind, temperature, ice cover, and chlorophyll-a concentrations. Penguins and other marine homeotherms are true inhabitants of the high seas. Depending on the season, the marine behaviour varies: during reproduction, penguins are central-place foragers, and must return regularly to their nest to feed their chicks. During the remainder of the year, there are no constraints and the birds travel large distances at sea. Breeding Humboldt penguins react to climatic change by varying their daily foraging range and dive duration. Similar to other representatives of the family Spheniscidae, Humboldt penguins avoid food shortages by migrating into more productive marine areas. Navigational clues such as daylength, sea surface temperature, local wind direction and olfaction might provide important aids in finding patchily distributed prey in the open ocean. DMS, a chemical compound produced by decaying algae, seems to be a further clue that indirectly points the way to feeding areas. PMID- 16351848 TI - Adaptation of Arctic and Antarctic ice metazoa to their habitat. AB - Sea ice is a unique habitat in polar seas. A diverse assemblage of plants and animals lives in its interior parts and at the ice-water interface. Their distribution is to a large extent controlled by abiotic parameters such as light, salinity and space, as well as food availability. In both the Arctic and Antarctic, the highest metazoan concentrations occur mostly in the bottom centimetres of the sea ice. Dominant metazoans are nematodes, turbellarians, rotifers and crustaceans. The ice-water interface itself houses in addition to endemic amphipods migrants from both the ice and the pelagic realm. To survive with the environmental conditions of the sea ice habitat, the ice biota is adapted, specifically to seasonal salinity variations from below 5 to above 60 PSU. Sea ice metazoans feed mainly on the algae growing within the sea ice. The loss of habitat during ice melt periods can lead to substantial sedimentation of ice fauna to the sea floor, where it might act as food source for the benthos. PMID- 16351849 TI - Invasion history and genetic population structure of riverine macroinvertebrates. AB - Macroinvertebrate communities of large rivers have experienced dramatic species turnovers in the last decades, which still go on. The analysis of genetic population structure plays a central role in understanding and predicting these biological invasions. Two points of view are considered: the influence of the invasion history on the genetic structuring and the potential implications of genetic structure for future invasibility. Expectations about selectively neutral genetic variation in simple invasion models are compared to case studies of amphipods and Dreissena. The genetic patterns of one amphipod species of the Gammarus fossarum complex yield strong evidence for a stepwise regional colonisation. Other invasive amphipods show similar, but less clear genetic characteristics. Long-distance colonisation is probably common in Dreissena polymorpha, but subsequent downstream swamping of larvae obscures the expected genetic pattern by homogenising neighbouring populations. First, a brief review of general characteristics of riverine invasions is followed by a discussion of the utility of genetic tools for inferring taxon and source population identity. PMID- 16351850 TI - ZOOLOGY moving towards its goals. PMID- 16351851 TI - Cranial neural crest-cell migration in the direct-developing frog, Eleutherodactylus coqui: molecular heterogeneity within and among migratory streams. AB - Direct development is a specialized reproductive mode that has evolved repeatedly in many different lineages of amphibians, especially anurans. A fully formed, albeit miniature adult hatches directly from the egg; there is no free-living larva. In many groups, the evolution of direct development has had profound consequences for cranial development and morphology, including many components that are derived from the embryonic neural crest. Yet, the developmental bases of these effects remain poorly known. In order to more fully characterize these changes, we used three molecular markers to analyze cranial neural crest-cell emergence and migration in the direct-developing frog, Eleutherodactylus coqui: HNK-1 immunoreactivity, Dlx protein expression, and cholinesterase activity. Our study validates and extends earlier results showing that the comprehensive changes in embryonic cranial patterning, differentiation, and developmental timing that are associated with direct development in Eleutherodactylus have not affected gross features of cranial neural crest biology: the relative timing of crest emergence and the number, configuration and identity of the principal migratory streams closely resemble those seen in metamorphic anurans. The three markers are variably expressed within and among neural crest-cell populations. This variation suggests that determination of cranial neural crest-cells may already have begun at or soon after the onset of migration, when the cells emerge from the neural tube. It is not known how or even if this variation correlates with differential cell lineage or fate. Finally, although HNK-1 expression is widely used to study neural crest migration in teleost fishes and amniotes, E. coqui is the only amphibian known in which it effectively labels migrating neural crest-cells. There are not enough comparative data to determine whether this feature is functionally associated with direct development or is instead unrelated to reproductive mode. PMID- 16351852 TI - Water fleas (Daphnia magna) provide a separate ventilatory mechanism for their brood. AB - The planktonic filter feeder Daphnia magna depends on a steady oxygen supply by convection. In the ventral carapace chamber, this convection is established by the feeding current which is generated by the movement of the thoracic limbs. The present study revealed that this movement can cause an additional flow of medium which passes through the brood chamber of the animal. To visualise this current, ink or fluorescent microspheres were released by a microcapillary near the posterior opening of the brood chamber. The tracks of these probes were monitored by video microscopy. Digital motion analysis was used for the determination of flow velocity and flow rate. Ambient medium entered the brood chamber at the abdomen of the animal and moved then to the anterior end of the brood chamber before entering the ventral carapace chamber. Two horizontal lamellae, which are attached at both sides of the trunk and project laterally to contact the carapace walls, almost completely separate the dorsal brood chamber from the ventral carapace chamber. Water can only pass these barriers through small depressions in these lamellae at the level of the 3rd and 4th appendages. Female daphnids with embryos at late developmental stages showed more rapid water currents in the brood chamber than those with younger embryos. Moreover, animals showed higher flow rates when exposed to hypoxic conditions. As the oxygen uptake rate of older embryos is approximately three times higher than that of younger embryos, the enhanced brood chamber current could improve the oxygen availability for both the mother and its brood under conditions of reduced oxygen availability. PMID- 16351853 TI - Fiber arrangement of the intestinal smooth muscle layers in ammocoetes of Lampetra japonica. AB - The three-dimensional arrangement of the intestinal smooth muscle in the ammocoetes of the lamprey (Lampetra japonica) was examined by scanning electron microscopy (SEM) after removal of the intestinal mucosa. In cross section of the posterior midgut, its wall was composed of the parietal wall and the typhlosolar wall of the spiral fold, lining a horseshoe-shaped space, and had two distinct muscle layers. The fiber extensions of the muscle layers in the two parts of the wall were reversed; internal longitudinal and external circular in the parietal wall, but internal circular and external longitudinal in the typhlosolar wall. The positional exchange of the two layers occurred within the transitional area from the parietal wall to the typhlosolar wall, where an interlacing texture of longitudinal and circular braids of fibers was observed. Furthermore, the external fibers of the longitudinal braid extended successively into the circular braid until the longitudinal braid disappeared. However, any fibrous transition from the circular braid into the longitudinal braid in the typhlosolar wall was not clear in the present study. The internal location of the longitudinal layer at the parietal wall may be optimal for its main function of contracting the intestinal tract longitudinally. In addition, the external (to be precise, the internal to the hematopoietic tissue) longitudinal muscle layer in the typhlosolar wall may play an important role in saving and squeezing out blood into the cardinal intestinal vein by longitudinal contraction of the elongated adjacent hematopoietic tissue mass. PMID- 16351854 TI - Immunocytochemical observations on the cornification of soft and hard epidermis in the turtle Chrysemys picta. AB - The process of cornification in the shell and non-shelled areas of the epidermis of the turtle Chrysemys picta was analyzed by light and ultrastructural immunohistochemistry for keratins, filaggrin and loricrin. Beta-keratin (hard keratin) was only present in the corneus layer of the plastron and carapace. The use of a beta-keratin antibody, developed against a specific chick scale beta keratin, demonstrated that avian and reptilian hard keratins share common amino acid sequences. In both, shelled and non-shelled epidermis, acidic alpha keratin (AE1 positive) was limited to tonofilament bundles of the basal and suprabasal layer, while basic keratin (AE3 positive) was present in basal, suprabasal, and less intensely, pre-corneus layers, but tended to disappear in the corneus layer. The AE2 antibody, which in mammalian epidermis recognizes specific keratins of cornification, did not stain turtle shell but only the corneus layer of non shelled (soft) epidermis. Two and four hours after an injection of tritiated histidine, the labelling was evenly distributed over the whole epidermis of both shelled and non-shelled areas, but was absent from the stratum corneum. In the areas of growth at the margin of the scutes of the shell, the labelling increased in precorneus layers. This suggests that histidine uptake is only related to shell growth and not to the production of a histidine-rich protein involved in keratinization. No filaggrin-like and loricrin-like immunoreactivity was seen in the carapace or plastron epidermis. However, in both proteins, some immunoreactivity was found in the transitional layer and in the lower level of the corneus layer of non-shelled areas. Loricrin- and filaggrin-like labelling was seen in small organelles (0.05-0.3 mum) among keratin bundles, identified with mucous-like granules and vesicular bodies. These organelles, present only in non-shelled epidermis, were more frequent along the border with the corneus layer, and labelling was low to absent in mature keratinocytes. This may be due to epitope masking or degradation. The immunolabelling for filaggrin was seen instead in the extracellular space among mature keratinocytes, over a material previously identified as mucus. The possibility that this labelling identified some epitopes derived from degraded portions of a filaggrin-like molecule is discussed. The present study suggests that proteins with some filaggrin- and loricrin-immunoreactivity are present in alpha-keratinocytes but not in beta keratin cells of the shell. PMID- 16351855 TI - An integrative model to predict maximum O2 uptake in animals with central vascular shunts. AB - Gas exchange in the tissues and lungs is accurately described by simple equations, and mathematical models of O(2) transport play a paramount role in the conceptual understanding of respiratory physiology. In the present paper, we review an integrative approach to describe maximum oxygen uptake in animals with tidally ventilated lungs. Further, we expand the analysis by including a central vascular shunt compartment to mimic the cardiovascular anatomy of embryonic birds and mammals as well as many ectothermic vertebrates. Finally, we predict the effects of right-to-left and left-to-right cardiac shunts on maximal oxygen uptake and present a new hypothesis that relates the interaction of metabolic rates and central shunts in air breathing ectothermic vertebrates. PMID- 16351856 TI - A skeletochronological study of age and growth in relation to adult size in the water frog Rana epeirotica. AB - The age and growth of individuals in a population of the water frog Rana epeirotica, from northwestern Greece, were studied in 1998 and 1999 using skeletochronology. Analysis of the age structure was based on the enumeration of Lines of Arrested Growth (LAGs) in phalanges. Ages ranged from 1-5 years among 145 males and from 1-5 years among 173 females. The mean snout-vent length was 72.74 +/- 7.96 mm in males and 83.63 +/- 11.73 mm in females. The difference between the sexes in age and size was significant. Endosteal resorption may have eroded away the first LAG in only 9% of the studied individuals and was more frequent in females. Growth of adults was fitted by the Bertalanffy model. The growth coefficient (K) was 0.88 in males and 0.49 in females, reflecting faster male growth between metamorphosis and maturation. PMID- 16351857 TI - Response properties of diencephalic neurons to visual, acoustic and hydrodynamic stimulation in the goldfish, Carassius auratus. AB - We studied the responses to sensory stimulation of three diencephalic areas, the central posterior nucleus of the dorsal thalamus, the anterior tuberal nucleus of the hypothalamus, and the preglomerular complex. Units sensitive to acoustic (500 Hz tone burst), hydrodynamic (25 Hz dipole stimulus) and visual (640 nm light flash) stimuli were found in both the central posterior and anterior tuberal nucleus. In contrast, unit responses or large robust evoked potentials confined to the preglomerular complex were not found. In the central posterior nucleus, most units were unimodal. Many units responded exclusively to visual stimulation and exhibited a variety of temporal response patterns to light stimuli. In the anterior tuberal nucleus of the hypothalamus, most units responded to more than one modality and showed a stronger response decrement to stimulus repetitions than units in the central posterior nucleus. Our data suggest that units in the central posterior nucleus are primarily involved in the unimodal processing of sensory information whereas units in the anterior tuberal nucleus of the hypothalamus may be involved in multisensory integration. PMID- 16351858 TI - Testing amniote models of prey transport kinematics: a quantitative analysis of mouth opening patterns in lizards. AB - Two models have been proposed to describe the prey transport kinematics of terrestrial vertebrates (Bramble and Wake, 1985; Reilly and Lauder, 1990). The critical difference between the models is the presence or absence of a slow open II phase (SO-II) in the gape profile during mouth opening. Each of these models has been applied to lizards, however to date, lizard feeding kinematics have not been adequately quantified to assess the utility of these models for this clade. Neither model has been sufficiently tested due to the lack of a methodology to assess the specific differences between the models. We describe a method that uses explicit mathematical criteria to define the kinematic phases in tetrapod feeding. This "slope analysis& is used to precisely quantify and compare the transport kinematics of seven lizard species. Lizard transport kinematics were highly variable both within and across taxa. However, several common gape cycle patterns were identified. The predominant patterns were slow-fast opening (37.3%), fast opening only (22.9%) and slow opening only (21.2%). The most common pattern explicitly fits the prediction of the Reilly and Lauder model while the other two are similar to patterns observed in salamanders. Thus, lizards possess both the slow opening-fast opening pattern predicted for amniotes and the more primitive, simple opening pattern characteristic of more basal tetrapods. Plateau phases were found in only 12.8% of the profiles and only a fourth of these (3.4% of the total) explicitly fit the Bramble and Wake model (slow opening, plateau, fast opening) and two species never exhibited plateaus in their gape cycles. Thus, it is clear that the Bramble and Wake model is not supported as a generalized model for lizards or generalized tetrapods. PMID- 16351859 TI - The phylogeny of amphibian metamorphosis. AB - Frogs have one of the most extreme metamorphoses among vertebrates. How did this metamorphosis evolve? By combining the methods previously proposed by Mabee and Humphries (1993) and Velhagen (1997), I develop a phylogenetic method suited for rigorous analysis of this question. In a preliminary analysis using 12 transformation sequence characters and 36 associated event sequence characters, all drawn from the osteology of the skull, the evolution of metamorphosis is traced on an assumed phylogeny. This phylogeny has lissamphibians (frogs, salamanders, and caecilians) monophyletic, with frogs the sister group of salamanders. Successive outgroups used are temnospondyls and discosauriscids, both of which are fossil groups for which ontogenetic data are available. In the reconstruction of character evolution, an unambiguous change (synapomorphy) along the branch leading to lissamphibians is a delay in the lengthening of the maxilla until metamorphosis, in accordance with my previous suggestion (Reiss, 1996). However, widening of the interpterygoid vacuity does not appear as a synapomophy of lissamphibians, due to variation in the character states in the outgroups. From a more theoretical perspective, the reconstructed evolution of amphibian metamorphosis involves examples of heterochrony, through the shift of ancestral premetamorphic events to the metamorphic period, caenogenesis, through the origin of new larval features, and terminal addition, through the origin of new adult features. Other changes don't readily fit these categories. This preliminary study provides evidence that metamorphic changes in frogs arose as further modifications of changes unique to lissamphibians, as well as a new method by which such questions can be examined. PMID- 16351860 TI - Variation, plasticity and modularity in anuran development. AB - Although anuran development is generally thought to be relatively conservative, a great deal of variation is evident when different species are compared. This report summarizes the results of comparative analyses of different aspects of anuran development. These include differences in sequence and timing of developmental events, the effects of genome size, and the effects of different life history strategies on anuran embryogenesis. The results show that anuran development is plastic at the evolutionary level, and many changes can occur in the developmental processes of anurans throughout their evolution. Changes are apparently rapid, and are as common as cladogenic events. This evolutionary plasticity can be attributed to the modular nature of anuran development. Different modules can shift relative to one another in time or in space, creating variations in the observed developmental patterns. However, shifts in modules can occur even without having a significant effect on the ultimate outcome of the process. I discuss the implications of the modular nature of development on the evolution of anuran development, and of the group in general. PMID- 16351861 TI - Development in frogs with large eggs and the origin of amniotes. AB - The origin of the amniote egg is one of the most significant events in the evolution of terrestrial vertebrates. This innovation was probably driven by increased egg size, and to find potential parallels, we can examine the derived development of extant amphibians with large eggs. The embryo of the Puerto Rican tree frog, Eleutherodactylus coqui, exhibits an alteration of its fate map and a secondary coverage of its yolky cells, reflecting the large 3.5 mm egg. Comparable changes may have occurred with the derivation of an amniote pattern of development. Future investigations should focus on the molecular organization of the egg. In the model amphibian for development, Xenopus laevis, information for embryonic germ layers, the dorsal axis, and germ cells is stored mainly as localized RNAs at the vegetal pole of the egg. These localizations would likely be changed with increased egg size. A review of the orthologues of the key X. laevis genes raises the possibility that their activities are not conserved in other vertebrates. PMID- 16351862 TI - Development and evolution of lateral line placodes in amphibians I. Development. AB - Lateral line placodes are specialized regions of the ectoderm that give rise to the receptor organs of the lateral line system as well as to the sensory neurons innervating them. The development of lateral line placodes has been studied in amphibians since the early 1900s. This paper reviews these older studies and tries to integrate them with more recent findings. Lateral line placodes are probably induced in a multistep process from a panplacodal area surrounding the neural plate. The time schedule of these inductive processes has begun to be unravelled, but little is known yet about their molecular basis. Subsequent pattern formation, morphogenesis and differentiation of lateral line placodes proceeds in most respects relatively autonomously: Onset and polarity of migration of lateral line primordia, the type, spacing, size and number of receptor organs formed, as well as the patterned differentiation of different cell types occur normally even in ectopic locations. Only the pathways for migration of lateral line primordia depend on external cues. Thus, lateral line placodes act as integrated and relatively context-insensitive developmental modules. PMID- 16351863 TI - A high standard metabolic rate constrains juvenile growth. AB - The allocation of energy to various components of an individual's energy budget is often viewed as a competitive process. As such, a tradeoff may exist between production (growth) and maintenance metabolism. One view of a potential tradeoff, termed "the principle of allocation", suggests that individuals with lower maintenance metabolic expenditures may have higher growth rates. To determine whether such a tradeoff exists, I analyzed the relationship between growth rate and maintenance metabolism of 225 juvenile snapping turtles housed in the laboratory. I measured growth from hatching to 6 months of age, and then measured oxygen consumption and calculated standard metabolic rate. Mean growth rate was 0.19 g d(-) and mean standard metabolic rate (SMR) was 1.41 kJ d(-). Maintenance metabolism and growth were negatively correlated after both were adjusted for body mass. The results support the "principle of allocation" theory: individuals with higher standard metabolic rates tended to have low growth rates. PMID- 16351864 TI - Post-cranial prey transport mechanisms in the black pinesnake, Pituophis melanoleucus lodingi: an x-ray videographic study. AB - Most previous studies of snake feeding mechanisms have focused on the functional morphology of the highly specialized ophidian jaw apparatus. Although some of these studies have included observations of post-cranial movements during feeding, the functional roles of these movements have remained poorly understood. In this study, we used x-ray videography to examine post-cranial prey transport mechanisms in a colubrid snake, Pituophis melanoleucus lodingi. We found that prey transport in this species progresses through four distinct phases, three of which are characterized by either undulatory or concertina-like movements of the anterior portion of the trunk. In the first phase of transport (the oral phase), unilateral movements of the jaws are used to pull the head forward around the prey. In the second phase (the orocervical phase), unilateral jaw movements continue, but are augmented by concertina-like movements of the anterior portion of the trunk. In the third phase (the cervical phase), prey transport occurs exclusively through concertina-like movements of the neck. Finally, in the fourth phase (the thoracic phase), prey is transported to the stomach via undulatory movements of the trunk. Our observations of feeding behavior in a phylogenetically diverse sample of fourteen other snake species demonstrate that similar post-cranial transport mechanisms are used by a wide variety of alethinophidian snakes that feed on large, bulky prey. PMID- 16351865 TI - Walking and running at resonance. AB - Humans and other animals can temporarily store mechanical energy in elastic oscillations, f(el), of body parts and in pendulum oscillations, f(p) = const sq.rt (g/L), of legs, length L, or other appendages, and thereby reduce the energy consumption of locomotion. However, energy saving only occurs if these oscillations are tuned to the leg propagation frequency f. It has long been known that f is tuned to the pendulum frequency of the free-swinging leg of walkers. During running the leg frequency increases to some new value f = f(r). We propose that in order to maintain resonance the animal, mass M, actively increases its leg pendulum frequency to the new value f(p,r) =const sq.rt (a(y)/L)=f(r), by giving its hips a vertical acceleration a(y)= F(y)/M. The pendulum frequency is increased if the impact force F(y) of the stance foot is larger than Mg, explaining the observation by Alexander and Bennet-Clark (1976) that F(v) becomes larger than Mg when animals start to run. Our model predictions of the running velocity U(r) as function of L, F(v), are in agreement with measurements of these quantities (Farley et al. 1993). The leg's longitudinal elastic oscillation frequency scales as f(el) = const sq.rt (k/M). Experiments by Ferris et al., (1998) show that runners adjust their leg's stiffness, k, when running on surfaces of different elasticity so that the total stiffness k remains constant. Our analysis of their data suggests that the longitudinal oscillations of the stance leg are indeed kept in tune with the running frequency. Therefore we conclude that humans, and by extension all animals, maintain resonance during running. Our model also predicts the Froude number of walking-running transitions, Fr = U(2)/gL approximately 0.5 in good agreement with measurements. PMID- 16351866 TI - Zoology www guide. PMID- 16351867 TI - Development and evolution of lateral line placodes in amphibians. - II. Evolutionary diversification. AB - The amphibian lateral line system develops from a series of lateral line placodes. The different phases of development from early induction, to pattern formation, differentiation, morphogenesis, and metamorphic fate were summarized in the first part of this review (Schlosser, 2002a). Here, a survey of the diversity of lateral line systems in amphibians is presented indicating that most phases of lateral line development have been subject to evolutionary changes. Several trends suggest important roles for both adaptive changes and internal constraints in amphibian lateral line evolution. Many of these trends involved the coordinated modification of different derivatives of lateral line placodes suggesting that these placodes are not only autonomous developmental modules, but also units of evolutionary variation that tend to be modified in a coherent and largely context-independent fashion. PMID- 16351868 TI - Cranial neural crest emergence and migration in the Mexican axolotl (Ambystoma mexicanum). AB - The timing and pattern of cranial neural crest cell emergence and migration in the Mexican axolotl, Ambystoma mexicanum, are assessed using scanning electron microscopy (SEM). Cranial neural crest cells emerge and begin to migrate at the time of neural fold closure and soon form three distinct streams. The most anterior (mandibular) stream emerges first, at the level of the mesencephalon. Cells in this stream migrate rostroventrally around the optic vesicle. The second (hyoid) and third (branchial) streams emerge in close succession at the level of the rhombencephalon and extend ventrolaterally. Cells forming the hyoid stream migrate rostral to the otic vesicle, whereas the branchial stream divides into two parallel streams, which migrate caudal to the otic vesicle. At later stages (stage 26 onwards) the cranial neural crest cells disperse into the adjacent mesoderm and can no longer be followed by dissection and SEM. The pattern of cranial neural crest emergence and migration, and division into migratory streams is similar to that in other amphibians and in the Australian lungfish (Neoceratodus forsteri). Emergence of crest cells from the neural tube, relative to the time of neural tube closure, occurs relatively late in comparison to anurans, but much earlier than in the Australian lungfish. These results establish a morphological foundation for studies in progress on the further development and fate of cranial neural crest cells in the Mexican axolotl, as well as for studies of the role of cranial neural crest in cranial patterning. PMID- 16351869 TI - Post-ovipositional development of the monocled cobra, Naja kaouthia (Serpentes: Elapidae). AB - External morphological development between oviposition and hatching of the monocled cobra, Naja kaouthia, is described. Ten developmental stages are diagnosed according to nine features. These include fusion of the body wall musculature along the ventral midline, appearance of the endolymphatic ducts, formation of the eyelid, and the appearance of scales on the head and body. Additional observations of the developing skull are made at four of these ten stages, based on cleared and stained heads. PMID- 16351870 TI - Worker piping triggers hissing for coordinated colony defence in the dwarf honeybee Apis florea. AB - Defending a large social insect colony containing several thousands of workers requires the simultaneous action of many individuals. Ideally this action involves communication between the workers, enabling coordinated action and a fast response. The Asian dwarf honeybee, Apis florea, is a small honeybee with an open nesting habit and a comparatively small colony size, features that leave them particularly exposed to predators. We describe here a novel defence response of these bees in which the emission of an initial warning signal from one individual ("piping") is followed 0.3 to 0.7 seconds later by a general response from a large number of bees ("hissing"). Piping is audible to the human ear, with a fundamental frequency of 384 +/- 31Hz and lasting for 0.82 +/- 0.35 seconds. Hissing is a broad band, noisy signal, clearly audible to the human observer and produced by slight but visible movements of the bees' wings. Hissing begins in individuals close to the piping bee, spreads rapidly to neighbours and results in an impressive coordinated crescendo occasionally involving the entire colony. Piping and hissing are accompanied by a marked decrease, or even cessation, of worker activities such as forager dancing and departures from the colony. We show that whereas hissing of the colony can be elicited without piping, the sequential and correlated piping and hissing response is specific to the presence of potential predators close to the colony. We suggest that the combined audio visual effect of the hissing might deter small predators, while the cessation of flight activity could decrease the risk of predation by birds and insects which prey selectively on flying bees. PMID- 16351871 TI - Arcus as a tensegrity structure in the arolium of wasps (Hymenoptera: Vespidae). AB - The unfolding of the hymenopteran attachment pad (arolium) may be achieved in two ways, hydraulic and mechanical. The first was confirmed in experiments with pressure applied to more proximal leg parts and on immersion in hypotonic solutions. Presumably, this way of unfolding does not play an important role for a living hornet. Mechanical unfolding was studied experimentally with the aid of a micromanipulator pulling the tendon of the musculus retractor unguis. Ablation experiments on different parts of the arolium indicated that the arcus is the most crucial element for mechanical unfolding. The shape of the arcus in closed and open conditions was measured using a 3D measurement microscope and reconstructed by means of 3D computer graphics. The arcus coils up upon being freed from the arolium tissues, and coils up even more after immersion into a 10% aqueous solution of NaOH. Geometrical models of the arcus are proposed, from which the rotational moment of elasticity is derived. Conformations and deformations of the arcus are quantified in order to explain its role in the folding and unfolding processes of the arolium. The diversity of approaches supports the idea that the arcus is a prestressed (tensegrity) structure providing immediate, soft, and graded transmission of forces during folding and unfolding action of the arolium. PMID- 16351872 TI - Plasma metabolites reflect seasonally changing metabolic processes in a long distance migrant shorebird (Calidris canutus). AB - Migrant birds have tightly scheduled annual cycles consisting of several distinct life cycle (sub-)stages such as reproduction, migration, moult and overwintering, each of which have specific metabolic requirements (e.g., fattening during migration, protein build-up during moult). This study examines changes in fat and protein metabolism during the annual cycle of body mass and moult over 1.5 years in a captive flock of an arctic-breeding shorebird, the red knot Calidris canutus islandica. 2-5 h after food withdrawal, plasma uric acid levels were still decreasing and beta-hydroxy-butyrate levels were low, indicating prolonged catabolism of dietary protein, probably linked with a conversion into lipids. Such a late-resorptive state is achieved much earlier in passerines, but only after several days in penguins and, thus, seems to depend on meal size or mass specific metabolic rate. Substages of body mass gain and high body mass were characterized by increased plasma triglyceride levels reflecting increased turnover of lipids, and low levels of the ketone body beta-hydroxy- butyrate, indicating that the bird is not short of glucose. The high uric acid levels during these substages indicated an increased breakdown of nutritional protein. During moult, plasma triglyceride levels were low, suggesting that lipids were less available than at other times of the year. It is concluded that plasma metabolite levels indicate the metabolic processes related to migratory fuelling and moult and the influence of exogeneous factors. PMID- 16351873 TI - Inter- and intraspecific variation in the use of marine food resources by three Cinclodes (Furnariidae, Aves) species: carbon isotopes and osmoregulatory physiology. AB - The avian genus Cinclodes (Furnariidae) includes species that inhabit both inland and marine shores. We compared the carbon isotopic composition and osmoregulatory capacities of field caught individuals of three Cinclodes species in Chile. Cinclodes nigrofumosus is a resident of coastal shores, whereas C. oustaleti and C. patagonicus inhabit both coastal and inland environments. The tissues of C. nigrofumosus exhibited distinctively marine delta(13)C values, whereas those of C. oustaleti and C. patagonicus were intermediate between marine and terrestrial values. The differences in carbon isotopic composition among these three species were paralleled by differences in osmoregulatory characteristics. The species that carbon isotopes revealed to be strictly marine, C. nigrofumosus, had relatively larger kidneys with a higher fraction of total renal tissue occupied by medullary cones than its congeners C. oustaleti and C. patagonicus. Cinclodes nigrofumosus individuals also produced more concentrated urine. In addition to interspecific differences in osmoregulation, we found intraspecific differences. Cinclodes nigrofumosus collected at an arid site with limited or no available fresh water exhibited larger kidneys and higher relative medullary thickness than individuals collected at a mesic site. Cinclodes nigrofumosus, like all passerines, lacks functional salt glands. This species appears to be unique among passerines in its ability to live in extreme arid coastal environments and to cope with a marine diet that imposes high osmotic loads. PMID- 16351874 TI - Earthworm leukocytes react with different mammalian antigen-specific monoclonal antibodies. AB - We identified conserved molecules (enzymes, peptides, cytokines) that might play a role in invertebrate innate immunity. We found these molecules by immunoserological and immunohistochemical methods in association with coelomocytes, leukocytes located in the coelomic cavity of the earthworm Eisenia foetida. We detected the enzyme Cu-Zn-superoxide-dismutase (SOD), cytokines (tumor necrosis factor-alpha, TNFalpha; transforming growth factor-alpha, TGFalpha; and alpha peptide hormone, thyreotrope stimulating hormone, TSH) in earthworm coelomocytes with monoclonal antibodies developed originally against human and/or mouse antigens. Three coelomocyte subpopulations were identified according to their form, size and granularity by microscopic and flow cytometric analysis. These cell populations showed different reactivity with antibodies against mammalian cell surface (CD) markers and different intracellular antigens. Two coelomocyte types showed cell surface positivity with anti-Thy-1 (CD90), CD24 and TNF-alpha antibodies. Strong cytoplasmic reaction was shown with anti-TNF alpha and anti-SOD mAbs and a weaker but unambiguous reaction with thyroid stimulating hormone (TSH) in two cell populations. The third population was negative for all of the monoclonal antibodies. Our flow cytometric results were confirmed by confocal microscopy both on the cell surfaces and intracellularly. PMID- 16351875 TI - Zoology www guide. PMID- 16351877 TI - Spider senses - technical perfection and biology. AB - This essay deals with sensory biology in a broad sense. It takes mechanosensory systems of spiders to illustrate a few basic issues. Particular attention is given to two aspects. 1. There is a remarkable "ingenuity" in the uptake and transformation of the adequate stimuli way out in the sensory periphery, which is reflected by an intimate relationship between the physical properties of the stimuli and the characteristics of the structures receiving and transforming them. We need to understand the details of this relationship in order to understand the relationship of an organism to its environment. 2. Sensory systems represent interfaces between the environment and behavior. As highly selective filters they have not evolved to provide abstract knowledge but to guide a particular behavior. The signals sent to the central nervous system are meaningful only in regard to their behavioral significance. - Some details of stimulus transformation in biological strain gauges (slit sensilla), airflow detectors (trichobothria) and touch receptors (tactile hairs) are described. Some of the refinement in the periphery is then meshed with the behavior of the whole organism. In this way the value shall be underlined of trying to understand reductionist details as building blocks of the complexity which enables an organism to behave in its own particular way in its species specific environment. PMID- 16351878 TI - The constraints of body size on aerodynamics and energetics in flying fruit flies: an integrative view. AB - Reynolds number and thus body size may potentially limit aerodynamic force production in flying insects due to relative changes of viscous forces on the beating wings. By comparing four different species of fruit flies similar in shape but with different body mass, we have investigated how small insects cope with changes in fluid mechanical constraints on power requirements for flight and the efficiency with which chemical energy is turned into aerodynamic flight forces. The animals were flown in a flight arena in which stroke kinematics, aerodynamic force production, and carbon dioxide release were measured within the entire working range of the flight motor. The data suggest that during hovering performance mean lift coefficient for flight is higher in smaller animals than in their larger relatives. This result runs counter to predictions based on conventional aerodynamic theory and suggests subtle differences in stroke kinematics between the animals. Estimates in profile power requirements based on high drag coefficient suggest that among all tested species of fruit flies elastic energy storage might not be required to minimize energetic expenditures during flight. Moreover, muscle efficiency significantly increases with increasing body size whereas aerodynamic efficiency tends to decrease with increasing size or Reynolds number. As a consequence of these two opposite trends, total flight efficiency tends to increase only slightly within the 6-fold range of body sizes. Surprisingly, total flight efficiency in fruit flies is broadly independent of different profile power estimates and typically yields mean values between 2-4%. PMID- 16351879 TI - The circadian system of Drosophila melanogaster and its light input pathways. AB - The fruit fly Drosophila melanogaster has been a grateful object for circadian rhythm researchers over several decades. Behavioral, genetic, and molecular studies in the little fly have aided in understanding the bases of circadian time keeping and rhythmic behaviors not only in Drosophila, but also in other organisms, including mammals. This review summarizes our present knowledge about the fruit fly's circadian system at the molecular and neurobiological level, with special emphasis on its entrainment by environmental light-dark cycles. The results obtained for Drosophila are discussed with respect to parallel findings in mammals. PMID- 16351880 TI - Learning in honeybees: from molecules to behaviour. AB - Studies in a variety of organisms as diverse as molluscs, insects, birds and mammals have shown that memories can exist in a variety of temporal domains ranging from short-term memories in the range of minutes to long-term memories lasting a lifetime. While transient covalent modifications of proteins underlie short-term memory, the formation of long-term memory requires gene expression and protein synthesis. Different intracellular signalling cascades have been implicated in distinct aspects of learning and memory formation. Little is known however, about how learning in intact animals is related to the modulation of these signalling cascades and how this contributes to distinct neuronal and behavioural changes in vivo. Associative learning in the honeybee provides the opportunity to study processes of memory formation by analysing its progression through different phases, across levels of behaviour, neural circuits, and cellular signalling pathways. The findings reveal evidence that various cellular signalling pathways in the neuronal circuit of distinct brain areas play a role in different processes during learning and memory formation. PMID- 16351881 TI - Animal learning and memory: an integration of cognition and ecology. AB - A wonderfully lucid framework for the ways to understand animal behaviour is that represented by the four 'whys' proposed by Tinbergen (1963). For much of the past three decades, however, these four avenues have been pursued more or less in parallel. Functional questions, for example, have been addressed by behavioural ecologists, mechanistic questions by psychologists and ethologists, ontogenetic questions by developmental biologists and neuroscientists and phylogenetic questions by evolutionary biologists. More recently, the value of integration between these differing views has become apparent. In this brief review, we concentrate especially on current attempts to integrate mechanistic and functional approaches. Most of our understanding of learning and memory in animals comes from the psychological literature, which tends to use only rats or pigeons, and more occasionally primates, as subjects. The underlying psychological assumption is of general processes that are similar across species and contexts rather than a range of specific abilities. However, this does not seem to be entirely true as several learned behaviours have been described that are specific to particular species or contexts. The first conspicuous exception to the generalist assumption was the demonstration of long delay taste aversion learning in rats (Garcia et al., 1955), in which it was shown that a stimulus need not be temporally contiguous with a response for the animal to make an association between food and illness. Subsequently, a number of other examples, such as imprinting and song learning in birds (e.g., Bolhuis and Honey, 1998; Catchpole and Slater, 1995; Horn, 1998), have been thoroughly researched. Even in these cases, however, it has been typical for only a few species to be studied (domestic chicks provide the 'model' imprinting species and canaries and zebra finches the song learning 'models'). As a result, a great deal is understood about the neural underpinnings and development of the behaviour, but substantially less is understood about interspecific variation and whether variation in behaviour is correlated with variation in neural processing (see review by Tramontin and Brenowitz, 2000 but see ten Cate and Vos, 1999). PMID- 16351882 TI - Precognitive and cognitive elements in sound localization. AB - Sound localization behavior is of great importance for an animal's survival. To localize a sound, animals have to detect a sound source and assign a location to it. In this review we discuss recent results on the underlying mechanisms and on modulatory influences in the barn owl, an auditory specialist with very well developed capabilities to localize sound. Information processing in the barn owl auditory pathway underlying the computations of detection and localization is well understood. This analysis of the sensory information primarily determines the following orienting behavior towards the sound source. However, orienting behavior may be modulated by cognitive (top-down) influences such as attention. We show how advanced stimulation techniques can be used to determine the importance of different cues for sound localization in quasi-realistic stimulation situations, how attentional influences can improve the response to behaviorally relevant stimuli, and how attention can modulate related neural responses. Taken together, these data indicate how sound localization might function in the usually complex natural environment. PMID- 16351883 TI - Development of renal function. AB - The mammalian metanephric kidney develops following a general principle of organogenesis of epithelial organs, i.e., along the tree-like structure of an arborizing ductal system (the ureteric bud and cortical collecting duct). In parallel, the proximal portions of the uriniferous tubule develop by mesenchymal to-epithelial transition of the neighbouring mesenchyme. On one hand, vectorial transport systems in nephrogenesis should be functional at the onset of glomerular filtration in any of the newly formed nephron generations to prevent loss of salt, water and metabolites. On the other hand, developing nephron epithelia must serve the needs of organ-formation such as cell proliferation and fluid-secretion for morphogenic purposes. This review intends to summarize current data and concepts on the development of renal epithelial functions with an emphasis on ion channels. Current model systems are introduced, such as ureteric bud cell monolayer culture, in vitro nephron culture, HEK293 cell culture, and the dissection of tubular cells for direct analysis. The current data on the developmental expression and functions of ENaC Na(+) channels, the CFTR, ClC-2 Cl(ndash;) channels, L-type Ca(2+) channels, P2 purinoceptors, and the Kir6.1/SUR2, ROMK (Kir1.1), and Kv K(+) channels are presented. PMID- 16351884 TI - Diversity of Anisoptera (Odonata): infering speciation processes from patterns of morphological diversity. AB - With roughly 2500 described species Anisoptera are among the species-poor suborders within insects. However, morphological and ecological variability are truly impressive. Anisoptera are classified into about 15 families of variable species richness. In this analysis phylogenetic research is integrated with comparative approaches to investigate possible explanations of differential speciation rates within this suborder. A short review of phylogenetic work based on morphological characters is compared to published molecular phylogenies. Sistergroup comparisons are used to elucidate whether a) sexual selection, b) duration of life cycles, or c) differentiation in body size, have had a detectable effect on speciation rate. In all three analyses effects of distributional range and latitudinal distribution were controlled. These analyses suggest sexual selection promotes speciation and an increase in body size is positively correlated with speciation rate. The evolutionary significance of these results is discussed and experimental approaches that should advance our understanding of anisopteran diversity are suggested. PMID- 16351885 TI - Causes and consequences of biological diversity in soil. AB - There is a vast diversity of organisms that live in the soil, and the activities of the total soil biota, together with the diverse forms and functions of plant roots, have critical roles in soil functioning. In this paper I discuss the likely determinants of soil diversity and also comment on recent studies that have explored whether or not there is a relationship between soil organism diversity and ecosystem function. There is little evidence to suggest that soil diversity is regulated in a predicable fashion by competition or disturbance; rather it is attributed to the nature of the soil environment, in that soil offers an extremely heterogeneous habitat, both spatially and temporally, proving unrivalled potential for niche partitioning, or resource or habitat specialisation, thereby enabling co-existence of species. Most evidence that is available suggests that there is no predictable relationship between diversity and function in soils, and that ecosystem properties are governed more by individual traits of dominant species, and by the extraordinary complexity of biotic interactions that occur between components of soil food webs. There is evidence of redundancy in soil communities with respect to soil functions, but the scale of effect of changes in soil diversity on process rates depends on which species are removed from the community and the degree to which remaining species can compensate. As in aboveground communities, therefore, it would appear that species traits and changes in species composition, and alterations in the nature of the many important species interactions that occur in soil, are likely to be the main biotic control of ecosystem function. In view of this, consideration of these important biotic interactions and their sensitivity to environmental change must be a key priority for future research. PMID- 16351887 TI - The distribution of putative nitric oxide releasing neurones in the locust abdominal nervous system: a comparison of NADPHd histochemistry and NOS immunocytochemistry. AB - Nitric oxide is well established as a signalling molecule in the nervous system of vertebrates and invertebrates. In this study we evaluate the usefulness of NADPHdiaphorase histochemistry and immunocytochemistry for detecting the presence of nitric oxide synthase in locusts. We describe the distribution of putative nitric oxide releasing neurones and stained neuropiles in the locust ventral nerve cord, in particular the abdominal ganglia and abdominal neuromeres. NADPHdiaphorase histochemistry revealed prominent staining in all neuropilar regions and a specific distribution pattern of stained cell bodies in all examined ganglia. Nitric oxide synthase immunocytochemistry, using a commercially available universal antibody, labelled cells in corresponding positions within the ganglia. This was confirmed by double labelling of alternate sections. Western blot analysis demonstrated that in locusts this universal NOS-antibody binds to a protein of similar size to nitric oxide synthase identified in other insect species. The antibody also labelled axons in most peripheral nerves of all examined ganglia, whereas NADPHdiaphorase histochemistry only revealed such stained fibres within peripheral nerves in some preparations, because they may have been masked by intense background staining. We therefore conclude that nitric oxide synthase-immunocytochemistry and NADPHd histochemistry are both good markers for the presence of nitric oxide synthase in the locust ventral nerve cord, and that nitric oxide may be used as a signalling molecule by efferent neurones in locusts. PMID- 16351888 TI - Maneuvering in juvenile carcharhinid and sphyrnid sharks: the role of the hammerhead shark cephalofoil. AB - The peculiar head morphology of hammerhead sharks has spawned a variety of untested functional hypotheses. One of the most intuitively appealing ideas is that the anterior foil acts, as in canard-winged aircraft, to increase maneuverability. We tested this hypothesis by determining whether juveniles of two hammerhead species (Sphyrna tiburo and S. lewini) turn more sharply, more often, and with greater velocity than a juvenile carcharhinid shark (Carcharhinus plumbeus). Although the hammerheads were more maneuverable, further investigation revealed that they do not roll their body during turns, suggesting that the cephalofoil does not act as a steering wing. We also show that hammerhead sharks demonstrate greater lateral flexure in a turn than carcharhinids, and that this flexibility may be due to cross sectional shape rather than number of vertebrae. PMID- 16351889 TI - Myoseptal architecture of sarcopterygian fishes and salamanders with special reference to Ambystoma mexicanum. AB - During axial undulatory swimming in fishes and salamanders muscular forces are transmitted to the vertebral axis and to the tail. One of the major components of force transmission is the myoseptal system. The structure of this system is well known in actinopterygian fishes, but has never been addressed in sarcopterygian fishes or salamanders. In this study we describe the spatial arrangement and collagen fiber architecture of myosepta in Latimeria, two dipnoans, and three salamanders in order to gain insight into function and evolution of the myoseptal system in these groups. Salamander myosepta lack prominent cones, and consist of homogenously distributed collagen fibers of various orientations that never form distinct tendons. Fiber orientations are difficult to homologize with those of fish myosepta. The myosepta of Latimeria and dipnoans (Protopterus and Neoceratodus) illustrate that major changes in architecture occurred in the sarcopterygian clade (loss of horizontal septum), in the rhipidistian (dipnoans + tetrapods) clade (loss of epineural and epipleural tendon), and in tetrapods (loss of lateral tendons and myoseptal folding). When compared to fishes, the myosepta of wholly aquatic salamanders (Ambystoma mexicanum, Amphiuma tridactylum, Necturus maculosus) do not have the lateral tendons we suppose serve to transfer muscular forces posteriorly. We propose that alternative structures (most conspicuously present in Ambystoma) perform this function: posteriorly the relative amount of connective tissue increases considerably, and myosepta are disintegrated to horizontal lamellae of connective tissue. The structures thought to be involved in modulation of body stiffness in fishes during swimming are also absent in salamanders. Our data also have implications for the hypothesis that salamander hypaxial myosepta are designed to increase shortening amplification of the hypaxial muscle fibers. The posterior hypaxial myosepta of all three salamander species possess only mediolaterally directed collagen fibers, which would indeed amplify the shortening of the associated muscle. PMID- 16351890 TI - Feeding in Atractaspis (Serpentes: Atractaspididae): a study in conflicting functional constraints. AB - African fossorial colubroid snakes of the genus Atractaspis have relatively long fangs on short maxillae, a gap separating the pterygoid and palatine bones, a toothless pterygoid, and a snout tightly attached to the rest of the skull. They envenomate prey with a unilateral backward stab of one fang projected from a closed mouth. We combined structural reanalysis of the feeding apparatus, video records of prey envenomation and transport, and manipulations of live and dead Atractaspis to determine how structure relates to function in this unusual genus of snakes. Unilateral fang use in Atractaspis is similar to unilateral slashing envenomation by some rear-fanged snakes, but Atractaspis show no maxillary movement during prey transport. Loss of pterygoid teeth and maxillary movement during transport resulted in the inability to perform. 'pterygoid walk' prey transport. Atractaspis transport prey through the oral cavity using movement cycles in which mandibular adduction, anterior trunk compression, and ventral flexion of the head alternate with mandibular abduction and extension of head and anterior trunk over the prey. Inefficiencies in manipulation and early transport of prey are offset by adaptability of the envenomating system to various prey types in both enclosed and open spaces and by selection of prey that occupy burrows or tunnels in soil. Atractaspis appears to represent the evolutionary endpoint of a functional conflict between envenomation and transport in which a rear-fanged envenomating system has been optimized at the expense of most, if not all, palatomaxillary transport function. PMID- 16351891 TI - Histochemical and ultrastructural features of the lingual epithelium of the rat snake (Elaphe climacophora). AB - The histological, ultrastructural and histochemical characteristics of the lingual epithelium of the rat snake (Elaphe climacophora) were investigated by light and transmission electron microscopy. The cells in the beta-layer of the epithelium of the bifurcated apex were filled with beta-keratin fibers and an amorphous matrix. Round projections covering the surface of the epithelial cells, namely, microfacets which contained pale granules, were clearly visible on the outer faces of Oberhautchen cells on the epithelium, and they were identified as fine granules filled with lipid. These granules might play an important role as a coating on the surface of the bifurcated lingual apex. The lipid on the surface of the lingual apex might also serve to trap and retain odorant molecules. Keratohyalin-like granules were distributed within the a-layer of the epithelium of the bifurcated apex of the tongue in the resting phase and cellular interdigitation was well developed in this region. Evidence of a shedding line was apparent under the light microscope in the cleft between outer and inner epithelial generations. The epithelial surface of the body of the tongue appeared suitable for retention of odorant and other molecules. PMID- 16351892 TI - Carpal ontogeny in Monodelphis domestica and Caluromys philander (Marsupialia). AB - Carpal bones have experienced numerous changes during marsupial evolution, even though their diversity and development remain poorly studied. The aim of this work was to document adult form and the pattern of mesenchymal tissue condensation and formation of chondrification and ossification centers in the hand of two marsupials. Two fundamental questions were asked: whether the loss of embryonic precursors was associated with the loss of adult elements, or whether there were developmental signs of ancestral mammalian elements that have been fused or lost in marsupial taxa. We were also interested to find out whether there is sexual dimorphismus in the carpals, as has been reported for some didelphids. Histological sections, cleared and stained specimens and macerated skeletons representing an ontogenetic series of Monodelphis domestica were used to document carpal development. Comparisons were made with perinatal stages of Caluromys philander and with adult specimens of other marsupials. A prenatal M. domestica in the 13th day after conception has a cell condensation that because of its position is homologized with a centrale, which is at birth already lost or fused. Neonatal M. domestica and C. philander have the number and arrangement of their adult carpal anatomy. Trapezium and trapezoid start ossification later than most other carpals, while pisiform and prepollex are the last to do so. Adult males of M. domestica have relatively larger and more robust pisiforms, compared to other carpals, than females. This sexual dimorphism develops relatively late as it was not recorded in male specimens around 160 days old. An extra sesamoid bone located just distal to the radius and proximo-palmar to the scaphoid was recorded in specimens of C. philander, C. derbianus and Didelphis virginiana. PMID- 16351894 TI - Development of physiological regulatory systems: altering the timing of crucial events. AB - There is currently tremendous interest in how the physiology of individual animals changes and develops during ontogeny. One of the key areas is the extent to which the timing and/or rate of physiological development is fixed within an individual and to what extent can it be altered. We propose that plasticity in the timing of the onset of a particular physiological regulatory system during an individuals development be referred to as physiological heterokairy (to clearly distinguish this phenomenon from physiological heterochrony, which is an evolutionary pattern), and we marshal evidence for three different patterns of heterokairy: 1. altering relative position in the physiological itinerary; 2. altering overall rate of development per se and; 3. a combination of 1 and 2. Using these patterns as a starting point, we develop a framework for investigating physiological heterokairy which takes cognizance of the facts that multiple components of each regulatory system could appear at different times and multiple regulatory systems could come 'on-line' at different times. We finish by placing physiological heterokairy in the wider context of its ecological and evolutionary implications and its relationship to physiological genomics and heterochrony. PMID- 16351895 TI - The Hydra viridis/Chlorella symbiosis. Growth and sexual differentiation in polyps without symbionts. AB - To investigate interactions between the basal metazoan Hydra viridis and its symbiotic Chlorella algae, we generated aposymbiotic hydra lacking algae and compared them to symbiotic ones with regard to growth and sexual differentiation. Under standard feeding conditions aposymbiotic polyps proliferated similarly to symbiotic polyps. Under moderate and low feeding conditions asexual growth was reduced in polyps lacking algae, indicating that the symbionts supply nutrients to their hosts. In addition, the Chlorella symbionts had a strong influence on the sexual reproduction of Hydra viridis: in most cases female gonads were produced only when symbiotic algae were present. Spermatogenesis proceeded similarly in symbiotic and aposymbiotic polyps. Since during oogenesis symbionts are actively transferred from endodermal epithelial cells to the ectodermal oocytes, this oogenesis promoting role could indicate that the symbionts are critically involved in the control of sexual differentiation in green hydra. PMID- 16351896 TI - Self/nonself recognition in Cnidaria: contact to allogeneic tissue does not result in elimination of nonself cells in Hydra vulgaris. AB - Although Cnidaria have no specialised immune cells, some colonial forms possess a genetic system to discriminate between self and nonself. Allorecognition is thought to protect them from fusion with genetically different individuals and to prevent germ line parasitism. Surprisingly, when grafting tissue of two species of the solitary freshwater polyp Hydra, we found within the contact zone phagocytozing epithelial cells which selectively eliminated cells from the other species (Bosch and David, 1986). This led us to speculate that Hydra, which never undergoes "natural transplantation", can differentiate between self and nonself (Bosch and David, 1986). In a previous paper (Kuznetsov et al., 2002) we described that cells which accumulate in the contact region of these interspecies grafts are apoptotic and that apoptosis is induced by impaired cell matrix contact. Thus, observations in such interspecies grafts did not give hints concerning the presence of a discriminative allorecognition system. To clarify whether this fundamental aspect of immunity is present in these phylogenetically old animals, we examined epithelial interactions between different strains of Hydra vulgaris. Here, we show that contact to allogeneic tissue does not evoke any response in terms of phagocytosis and elimination of allogeneic cells. We, therefore, question Hydra's ability to discriminate between self and nonself and propose that, in contrast to colonial cnidarians, the solitary polyp Hydra has either lost or substantially reduced this ability. PMID- 16351897 TI - Environmental stress and fluctuating asymmetry in the grasshopper Chorthippus parallelus (Acrididae: Gomphocerinae). AB - We examined the effect of agrochemical pollution/fertilisation of the soil and climatic conditions at high altitudes on fluctuating asymmetry (FA) level in hindfemur and forewing lengths of the grasshopper Chorthippus parallelus (Zetterstedt). All traits and samples exhibited ideal FA. Forewings, which are less functionally significant in this species, generally exhibited greater FA than the more functionally significant femora. Forewing FA was also more affected by stress induced by climatic conditions at high altitudes than femur FA. The high altitude sample was the most asymmetric sample for both traits, followed by the two control samples. The two samples from agrochemically contaminated habitats showed the lowest asymmetry in femora and forewings of C. parallelus. These results suggest that high altitude imposes more stress on this grasshopper species than does agrochemical contamination although the stress response was more dramatic in the less functional trait. PMID- 16351898 TI - Feeding in flow extremes: dependence of cirrus form on wave-exposure in four barnacle species. AB - Wave-exposure influences the form of many organisms. Curiously, the impact of flow extremes on feeding structures has received little attention. Barnacles extend feather-like legs to feed, and prior work revealed a highly precise association between leg length and water velocity in one species. To assess the generality of this flow-dependence, we quantified variation in four leg traits (ramus length, ramus diameter, seta length, and intersetal spacing) in four intertidal barnacles (Balanus glandula, Chthamalus dalli, Semibalanus cariosus, Pollicipes polymerus) over a wave-exposure gradient in the North-Eastern Pacific. All species exhibited a negative allometric relation between leg length and body mass. Proportionally longer feeding legs may permit smaller barnacles to avoid lower flow and particle flux associated with boundary layers. Although coefficients of allometry did not vary with wave-exposure, form differences among wave-exposures were substantial. Depending on the species, acorn barnacles of the same size from protected shores had feeding legs that were 37-80% longer and 18 25% thinner, and setae that were 36-50% longer and up to 25% more closely spaced, than those from exposed shores. Differences were less pronounced for the gooseneck barnacle, P. polymerus. Moreover, in situ water velocity explained an impressive percentage of overall leg-length variation: 92% in B. glandula, 67% in C. dalli, 91% in S. cariosus, and 92% in P. polymerus. Clearly, both size and shape of barnacle feeding legs respond to local flow conditions. This response appears widespread--across two orders of thoracican barnacles, Pedunculata and Sessilia, and two superfamilies of acorn barnacles (Balanoidea and Chthamaloidea) -and likely adaptive. Longer rami and setae would yield a larger feeding area in low flow, whereas shorter, stouter rami with shorter setae would be less vulnerable to damage in high flow. Finally, the proportionally most variable species was abundant in the widest range of habitats, suggesting that increased plasticity may permit a wider niche breadth. PMID- 16351899 TI - The parasympathetic nervous system and its influence on heart rate in torpid western pygmy possums, Cercatetus concinnus (Marsupialia: Burramyidae). AB - We investigated the effect of parasympathetic antagonism on the patterns of heart rate during torpor in the western pygmy possum Cercatetus concinnus (Marsupialia: Burramyidae). This is the first study to examine the influence of the autonomic nervous system on cardiac function in a metatherian hibernator. During torpor, antagonism of the parasympathetic nervous system eliminated the ventilatory tachycardia, variability in instantaneous heart rate, and increased the overall heart rate. These findings are consistent with previous studies on other mammalian heterotherms, which have shown that the parasympathetic nervous system is responsible for these patterns in heart rate. During extended bouts of torpor (2 to 3 days) the ventilatory tachycardia persisted throughout each bout, which indicates that the parasympathetic nervous system remained functional during that time. It has been suggested that the progressive removal of autonomic tone is characteristic of deep steady-state hibernation. There is no evidence to suggest that such a state was going to be reached in the possums in this study. To date there is little evidence that clearly demonstrates a physiological basis for the distinction between shallow, daily torpor and deep hibernation. PMID- 16351900 TI - Ingestion of juniper foliage reduces metabolic rates in woodrat (Neotoma) herbivores. AB - Ingestion of plant secondary compounds by herbivores is predicted to increase resting or basal metabolic rates. We tested this hypothesis with two species of woodrat herbivores, Neotoma stephensi and Neotoma albigula, consuming diets of juniper (Juniperus monosperma), which is rich in plant secondary compounds. In nature, N. stephensi specializes on juniper, whereas N. albigula consumes a variety of plant species including juniper. We measured resting metabolic rates (RMR) of woodrats on control, 25% juniper and a treatment containing the maximum tolerable dose of juniper (50% juniper for N. albigula and 70% juniper for N. stephensi). Ingestion of a juniper diet resulted in decreased RMR in both species of woodrats. We propose several potential mechanisms for metabolic depression of Neotoma on juniper diets. Our novel results underscore the need for more studies utilizing plant-based diets to determine the general effect of plant secondary compounds on metabolic rates of herbivores. PMID- 16351901 TI - Quantification of flow during suction feeding in bluegill sunfish. AB - Nearly all aquatic-feeding vertebrates use some amount of suction to capture prey items. Suction prey capture occurs by accelerating a volume of water into the mouth and taking a prey item along with it. Yet, until recently, we lacked the necessary techniques and analytical tools to quantify the flow regime generated by feeding fish. We used a new approach; Digital Particle Image Velocimetery (DPIV) to measure several attributes of the flow generated by feeding bluegill sunfish. We found that the temporal pattern of flow was notably compressed during prey capture. Flow velocity increased rapidly to its peak within 20 ms of the onset of the strike, and this peak corresponded to the time that the prey entered the mouth during capture. The rapid acceleration and deceleration of water suggests that timing is critical for the predator in positioning itself relative to the prey so that it can be drawn into the mouth along with the water. We also found that the volume of water affected by suction was spatially limited. Only rarely did we measure significant flow beyond 1.75 cm of the mouth aperture (in 20 cm fish), further emphasizing the importance of mechanisms, like locomotion, that place the fish mouth in close proximity to the prey. We found that the highest flows towards the mouth along the fish midline were generated not immediately in front of the open mouth, but approximately 0.5 cm anterior to the mouth opening. Away from the midline the peak in flow was closer to the mouth. We propose that this pattern indicates the presence of a bow wave created by the locomotor efforts of the fish. In this scheme, the bow wave acts antagonistically to the flow of water generated by suction, the net effect being to push the region of peak flow away from the open mouth. The peak was located farther from the mouth opening in strikes accompanied by faster locomotion, suggesting faster fish created larger bow waves. PMID- 16351902 TI - Automatic life-long monitoring of individual insect behaviour now possible. AB - Automatic tracking and identification of individuals has the potential to revolutionize the study of insects, especially social insects, by opening up options for questions which could not be asked before. To achieve this we developed a reliable and cost-sensible RFID (Radio-Frequency Identification) based solution that automatically recognises a virtually unlimited number (18 x 10(18) possible ID numbers) of individual insects down to the size of bees and ants. The data are collected automatically for any desired time span (if interesting, up to the entire life of the individual), pre-processed and saved in a database for further analysis. The usage of database techniques allows parallel data processing with a virtually unlimited number of parameter connections. ID numbers can be linked to any simultaneously recorded parameters of interest, e.g. spatial and temporal information as shown here for a bumblebee colony. PMID- 16351903 TI - Role of chemical and visual cues in food recognition by leatherback posthatchlings (Dermochelys coriacea L). AB - We raised leatherback posthatchlings in the laboratory for up to 7 weeks to study the role of visual and chemical cues in food recognition and food-seeking behavior. Turtles were reared on a formulated (artificial gelatinous) diet and had no contact with test materials until experiments began. Subjects were presented with visual cues (a plastic jellyfish; white plastic shapes [circle, square, diamond] similar in surface area to the plastic model), chemical cues (homogenates of lion's mane jellyfish, Cyanea capillata; moon jellyfish, Aurelia aurita; and a ctenophore, Ocyropsis sp., introduced through a water filter outflow), and visual and chemical cues presented simultaneously. Visual stimuli evoked an increase in swimming activity, biting, diving, and orientation toward the object. Chemical cues elicited an increase in biting, and orientation into water currents (rheotaxis). When chemical and visual stimuli were combined, turtles ignored currents and oriented toward the visual stimuli. We conclude that both cues are used to search for, and locate, food but that visual cues may be of primary importance. We hypothesize that under natural conditions turtles locate food visually, then, as a consequence of feeding, associate chemical with visual cues. Chemical cues then may function alone as a feeding attractant. PMID- 16351904 TI - Functional morphology and evolutionary origin of the three-part pharynx in nematodes. AB - The distinct morphological regions of the typical tripartite pharynx found in the nematode taxon Secernentea have distinctive functions. Besides the basic functions of sucking and pumping food against the pressure in the body cavity, the pharynx of Secernentea such as rhabditids serves two additional functions restricted to two pharyngeal subunits. The corpus traps bacteria behind the stoma and at its posterior end. The newly discovered pharyngeal pocket valve helps to trap particles behind the corpus in the rhabditid Poikilolaimus oxycercus and the cephalobid Acrobeles ciliatus (both Secernentea). The grinder of the terminal bulb serves for chewing trapped bacteria. The separated sites of trapping and chewing are connected by the isthmus that transports bacteria towards the grinder. It is likely that this complex feeding structure originated step by step from a two-part pharynx comprising a propharynx and the terminal bulb as in "Plectidae" (that probably include the closest relatives of the Secernentea within the "Adenophorea"). Analysis of video sequences of feeding rhabditids and plectids provided new data to reconstruct this transformation. Within the "Plectidae" two types of grinders occur. The first type or "parietinus type" has triangular chewing plates that can bulge medially and crush food particles. When they retract, new ingested particles are drawn into the grinder. The second type with more solid chewing plates called "butterfly valves" occurs in Ceratoplectus, Plectus parvus, and Wilsonema and can be homologized with the grinder in Secernentea ("Plectidae" is a paraphyletic taxon). Because butterfly valves cannot be retracted, the evolution of such valves required the evolution of an alternative mechanism to fill the grinder with bacteria. The differentiated closing pattern of the dilated pharynx lumen in Ceratoplectus, Plectus parvus, and Wilsonema can be interpreted as the first step in the development of a functional separation of trapping bacteria and of transporting them towards the grinder, which led to the morphologically discernible units of corpus and isthmus found in the Secernentea. PMID- 16351905 TI - Cruising specialists and accelerators--are different types of fish locomotion driven by differently structured myosepta? AB - Locomotor specialists, such as accelerators and cruisers, have clearly differing body designs. For physical reasons these designs are mutually exclusive, i.e. cruisers necessarily have poor accelerating capabilities and vice versa. For the first time, we examine whether differences in the anatomy of the musculo tendinous system of the trunk are present in addition to the differences in external body design. We investigated the myoseptal series of two closely related locomotor specialists, the cruiser Scomber scombrus and the accelerator Channa obscura, by microdissections combined with polarized light microscopy and histology. Our comparison includes 3D-morphology of myosepta, spatial arrangement and length of myoseptal tendons, their relation to red and white muscles, rostrocaudal changes in all these aspects and the musculo-tendinous system of the caudal fin. Regarding all these features, Channa has retained the plesiomorphic condition of its actinopterygian ancestor. In contrast, the derived morphology of Scomber is characterized by (i) lateral (LT) and myorhabdoid tendons (MT) that are lengthened to up to 20% of body length (compared to a maximum of 8.2% in Channa), (ii) posterior myoseptal cones that are subsequently linked by horizontal projections of merged LTs and MTs, (iii) an increased area of red muscle fibers that insert to LTs of myosepta, (iv) the reduction of epineural (ENTs) and epipleural tendons (EPTs) that connect backbone and skin, (v) specific caudal tendons that are identified to be serial homologues of LTs and MTs of more anterior myosepta, (vi) and a partial reduction of intrinsic caudal muscles. These results suggest the following functional adaptations in the cruiser Scomber. Red muscle forces may be transmitted through LTs and posterior cones to the prominent tendons of the caudal fin. The length of LTs and the intersegmental connections along the posterior cones may facilitate posterior force transmission and may be correlated with the long propulsive wavelength generally observed in cruising carangiform swimmers. Epineural and epipleural tendons are interpreted to minimize lateral backbone displacement during high body curvatures. This is consistent with the lack of these tendons in Scomber, because high body curvatures are not displayed in stiffer-bodied carangiform swimmers. It remains to be tested whether the specializations revealed in this initial study for Scomber represent general specializations of carangiform swimmers. Taking into account the geometry of myoseptal tendons and the horizontal septum we evaluate how local bending according to beam-theory can be generated by white or red muscle activity in Channa and Scomber. In both species, the musculo-tendinous anatomy of the caudal fin explains the functional asymmetry of the caudal fin that was experimentally revealed in previous studies. PMID- 16351906 TI - Spinning activity of the spider Trogloneta granulum (Araneae, Mysmenidae): web, cocoon, cocoon handling behaviour, draglines and attachment discs. AB - The spider species Trogloneta granulum, which in the wild lives inside scree slopes, builds three-dimensional orb webs. During egg-laying and egg sac building, the females stay with their dorsa down at the central part of the web. In this process, the hub is used as a platform. The threads of the hub are not incorporated into the silk cover of the egg sac. The silk wall of the egg sac is very thin, with all the silken threads constituting it having a uniform ultrastructure. The silk wall of the egg sac and the spinnerets are permanently linked by a dragline. Draglines produced by T. granulum affect the direction of movements of the female carrying its cocoon. Egg sacs are handled using draglines. The low number of piriform glands leads to the formation of very simple attachment discs, which fix the individual threads to the substratum. Thread bundles are attached to the substratum by means of accumulated attachment discs. PMID- 16351907 TI - The ear region of Latimeria chalumnae: functional and evolutionary implications. AB - The anatomy of Latimeria chalumnae has figured prominently in discussions about tetrapod origins. While the gross anatomy of Latimeria is well documented, relatively little is known about its otic anatomy and ontogeny. To examine the inner ear and the otoccipital part of the cranium, a serial-sectioned juvenile coelacanth was studied in detail and a three-dimensional reconstruction was made. The ear of Latimeria shows a derived condition compared to other basal sarcopterygians in having a connection between left and right labyrinths. This canalis communicans is perilymphatic in nature and originates at the transition point of the saccule and the lagena deep in the inner ear, where a peculiar sense end organ can be found. In most gnathostomes the inner ears are clearly separated from each other. A connection occurs in some fishes, e.g. within the Ostariophysi. In the sarcopterygian lineage no connections between the inner ears are known except in the Actinistia. Some fossil actinistians show a posteriorly directed duct lying between the foramen magnum and the notochordal canal, similar to the condition in the ear of Latimeria, so this derived character complex probably developed early in actinistian history. Because some features of the inner ear of Latimeria have been described as having tetrapod affinities, the problem of hearing and the anatomy of the otical complex in the living coelacanth has been closely connected to the question of early tetrapod evolution. It was assumed in the past that the structure found in Latimeria could exemplify a transitional stage in otic evolution between the fishlike sarcopterygians and the first tetrapods in a functional or even phylogenetic way. Here the possibility is considered that the canalis communicans does not possess any auditory function but rather is involved in sensing pressure changes during movements involving the intracranial joint. Earlier hypotheses of a putative tympanic ear are refuted. PMID- 16351908 TI - The ear of Latimeria chalumnae revisited. PMID- 16351911 TI - Evolution in Darwin's finches: a review of a study on Isla Daphne Major in the Galapagos archipelago. AB - This paper reviews research pertaining to the problem of speciation of the finches on the Galapagos archipelago carried out by assistants, several colleagues, Peter Grant and myself. I give a brief history of the radiation, examine the process of divergence by natural selection over time, and then consider the nature of the reproductive barrier to gene flow between closely related species. Fluctuating climatic conditions have produced a continuously changing ecological landscape and altered feeding conditions for the finches over the last 30 years. Finch populations tracked these changes by natural selection and evolutionary responses to the main events. At each event significant morphological change occurred from one generation to the next generation. As a consequence of these accumulated changes, the mean bill shape and body size of the Geospiza fortis and G. scandens populations differed markedly from 1973 to 2002. Song, a learned culturally transmitted trait, acted as a barrier to reproduction between these species. Rare incidences of misimprinting on song led to hybridization and introgression. Low levels of gene flow from one species to another increased genetic variation on which selection acted. Although the major driving force of diversification was ecological change, the process of diversification involved a subtle interplay between ecology, genetic evolution and learned culturally transmitted traits. An important message for conservation is that neither the environment nor species are fixed entities, therefore a wise strategy for conserving endangered species should keep them capable of further change. PMID- 16351912 TI - Behavioral genomics and the study of speciation at a porous species boundary. AB - Porous species boundaries are characterized by differential gene flow, where some regions of the genome experience divergent evolution while others experience the homogenizing effects of gene flow. If species can arise or remain distinct despite gene flow between them, speciation can only be understood on a gene by gene level. To understand the genetics of speciation, we therefore must identify the targets of selection that cause divergent evolution and identify the genetic architecture underlying such "speciation phenotypes". This will enable characterization of genomic regions that are "free to flow" between species, and those that diverge in the face of gene flow. We discuss this problem in the genus Laupala, a morphologically cryptic, flightless group of crickets that has radiated in Hawaii. Because songs are used in courtship and always distinguish close relatives of Laupala as well as species in sympatry, we argue that songs in Laupala are speciation phenotypes. Here, we present our approaches to identify the underlying genomic regions and song genes that differentiate closely related species. We discuss what is known about the genetic basis of this species difference derived from classic quantitative genetics and quantitative trait locus mapping experiments. We also present a model of the molecular expression of cricket song to assist in our goal to identify the genes involved in song variation. As most species are sympatric and exchange genes with congeners, we discuss the importance of understanding the genetic and genomic architecture of song as a speciation phenotype that must be characterized to identify differential patterns of gene flow at porous species boundaries. PMID- 16351913 TI - Why do species exist? Insights from sexuals and asexuals. AB - Why does life diversify into the more or less discrete entities we recognise as species? Two main explanations have been proposed: i) species are a consequence of adaptation to different ecological niches, ii) species are a consequence of sexual reproduction and reproductive isolation. Phylogenetic studies of case study groups can provide insights into the relative importance of divergent selection and isolation for speciation, but it can be difficult to infer causes of speciation unambiguously. The example of North American tiger beetles from the genus Cicindela is discussed. An alternative approach is to compare diversification between related sexual and asexual taxa to infer the relative importance of the two explanations. We outline expected patterns of diversification in sexual and asexual lineages under different scenarios using coalescent theory. Whether sexuals or asexuals diversify to a greater extent depends on the balance among various stages of diversification, particularly on the effects of sexual reproduction on rates of adaptive evolution. Rotifers offer a unique system to test these ideas, allowing comparison of patterns of genetic and functional morphological diversification in sexual (bdelloid) and asexual (monogonont) clades. PMID- 16351914 TI - The power of evo-devo to explore evolutionary constraints: experiments with butterfly eyespots. AB - Examples of adaptive radiation by Darwinian evolution will always enthral, and the theory of natural selection binds the whole of biology. But is selection all powerful? Evolutionary developmental biology is beginning to provide the background to understanding how the internal organisation of organisms can influence the tempo and direction of evolutionary change. Do the mechanisms that generate the phenotype channel, bias or limit morphological evolution? The formation of eyespots on the wings of the butterfly Bicyclus anynana is a process which is becoming comparatively well understood from the genetical, developmental and ecological perspectives. We have begun to combine this knowledge with artificial selection experiments to examine the rates at which morphological changes of a particular eyespot pattern in different directions can be achieved. Our initial selection experiment performed over twenty-five generations has shown that in contrast to predictions based on shared genetical and developmental properties, two butterfly eyespots on the same wing surface can be freely uncoupled from each other with respect to their pattern of relative sizes. This flexibility in development and in response to selection may occur because our base population is part of a lineage with a long legacy of natural selection building up genetic variation that enables independent behaviour for eyespot size, and thus it no longer reflects a tightly modular organisation of eyespots as may well have existed close to their evolutionary origin in basal Lepidoptera. Comparisons can be made between such descriptions of the potential for short-term changes in morphology in B. anynana and observed patterns of divergence among all extant species in this species-rich genus. Being able to map phenotypes onto genotypes via developmental processes provides a powerful basis for exploring genetical and developmental constraints. This will eventually lead to examples of evolutionary constraints that represent more than 'just-so' stories, and thus to a more balanced view of Darwinian evolution. PMID- 16351915 TI - Old trees, new trees--is there any progress? AB - The amount of comparative data for phylogenetic analyses is constantly increasing. Data come from different directions such as morphology, molecular genetics, developmental biology and paleontology. With the increasing diversity of data and of analytical tools, the number of competing hypotheses on phylogenetic relationships rises, too. The choice of the phylogenetic tree as a basis for the interpretation of new data is important, because different trees will support different evolutionary interpretations of the data investigated. I argue here that, although many problematic aspects exist, there are several phylogenetic relationships that are supported by the majority of analyses and may be regarded as something like a robust backbone. This accounts, for example, for the monophyly of Metazoa, Bilateria, Deuterostomia, Protostomia (= Gastroneuralia), Gnathifera, Spiralia, Trochozoa and Arthropoda and probably also for the branching order of diploblastic taxa ("Porifera", Trichoplax adhaerens, Cnidaria and Ctenophora). Along this "backbone", there are several problematic regions, where either monophyly is questionable and/or where taxa "rotate" in narrow regions of the tree. This is illustrated exemplified by the probable paraphyly of Porifera and the phylogenetic relationships of basal spiralian taxa. Two problems span wider regions of the tree: the position of Arthropoda either as the sister taxon of Annelida (= Articulata) or of Cycloneuralia (= Ecdysozoa) and the position of tentaculate taxa either as sister taxa of Deuterostomia (= Radialia) or within the taxon Spiralia. The backbone makes it possible to develop a basic understanding of the evolution of genes, molecules and structures in metazoan animals. PMID- 16351916 TI - Molecules, development and fossils in the study of metazoan evolution; Articulata versus Ecdysozoa revisited. AB - Two conflicting hypotheses of protostome relationships, Articulata and Ecdysozoa, are reviewed by evaluating the evidence in favor and against each one of them. Understanding early embryonic development and segmentation in non-arthropod non annelid protostomes seems crucial to the debate. New ways of coding metazoan matrices, avoiding ground-patterns and higher taxa, and incorporating fossil evidence seems the best way to avoid circular debates. Molecular data served as the catalyzer for the Ecdysozoa hypothesis, although morphological support had been implicitly suggested. Most molecular analyses published so far have shown some support for Ecdysozoa, whereas none has ever supported Articulata. Here, new analyses of up to four nuclear loci, including 18S rRNA, myosin heavy chain II, histone H3 and elongation factor 1-alpha are conducted to test the molecular support for Ecdysozoa, and, at least under some parameter sets, most data sets show a clade formed by the molting animals. In contrast, support for Articulata is not found under any analytical conditions. PMID- 16351917 TI - Sex, parasites and resistance--an evolutionary approach. AB - Immune systems are among the most diverse biological systems. An evolutionary arms race between hosts and rapidly evolving pathogens is supposed to be a reason for this diversity, and might explain why most eukaryotic hosts and parasites reproduce sexually. In this review, I will focus on possible benefits of sexual reproduction in hosts and parasites, using a model system consisting of a tapeworm and its two intermediate hosts, copepods and sticklebacks. We found that the hermaphroditic tapeworms can increase their infection success by reproducing sexually with a partner (outcrossing), instead of reproducing alone. The defence system of the copepods provides highly specific discrimination of antigenic characteristics of the tapeworms. This supports the finding that tapeworms benefit from outcrossing, but contradicts the conventional notion that the immune system of invertebrates, in contrast to vertebrates, is not able to react with specificity. Finally, sticklebacks seem to benefit from optimal diversity in their specific immune system. Previous studies showed that female sticklebacks prefer mates, which sire offspring with an optimal diversity in the MHC (genes involved in antigen presentation). We now found that these individuals suffer less from tapeworm infection. Furthermore, they are able to reduce the expression of an unspecific immune trait, thereby possibly avoiding harmful side effects of a highly activated, unspecific immune system. PMID- 16351918 TI - Intracellular parasitism: cell biological adaptations of parasitic protozoa to a life inside cells. AB - Several protozoan parasites evade the host's immune defence because most of their development takes place inside specific host cells. Only a few of these protozoa live within the host cell cytosol. Most parasites are sequestered within membrane bound compartments, collectively called 'vacuoles'. Recent advances in the cell biology of intracellular parasites have revealed fundamental differences in the strategies whereby such organisms gain entry into their respective host cells. These differences have important implications for host-parasite interaction and for nutrient acquisition by the parasite. Leishmania spp. take advantage of the phagocytic properties of their host cells and presumably contribute little to the uptake process. In contrast, apicomplexan parasites have developed highly specialised organelles, called micronemes and rhoptries, to actively invade a variety of nucleated cells and, in the case of Plasmodium falciparum, human erythrocytes. Following invasion, parasites use a multitude of strategies to protect themselves from the defence mechanisms of the parasitized cells. In addition, they induce novel pathways within the infected cell that allow a most efficient nutrient acquisition both from the host cell cytoplasm and from the extracellular environment. Parasite-induced changes of host cells are most apparent in erythrocytes infected with Plasmodium spp. Mammalian erythrocytes are deficient in de novo protein and lipid biosynthesis and, consequently, pathways which allow the transport of macromolecules and small solutes are established by metabolic activities of the parasite. Research into the cell biology of intracellular parasitism has identified fascinating phenomena some of which we are beginning to understand at a molecular level. They are fascinating because they allow insights into a very intimate interaction between two eukaryotic cells of entirely different phylogenetic origins. PMID- 16351919 TI - Parasitic worms: strategies of host finding, recognition and invasion. AB - Many parasitic worms enter their hosts by active invasion. Their transmission success is often based on a mass production of invasive stages. However, most stages show a highly specific host-finding behaviour. Information on host-finding mechanisms is available mainly for trematode miracidia and cercariae and for nematode hookworms. The larvae find and recognise their hosts, in some cases even with species specificity, via complex sequences of behavioural patterns with which they successively respond to various environmental and host cues. There is often a surprisingly high diversity of host-recognition strategies. Each parasite species finds and enters its host using a different series of cues. For example, different species of schistosomes enter the human skin using different recognition sequences. The various recognition strategies may reflect adaptations to distinct ecological conditions of transmission. Another question is how, after invasion, parasitic worms find their complex paths through their host's tissues to their often very specific microhabitats. Recent data show that the migrating parasite stages can follow local chemical gradients of skin and blood compounds, but their long-distance navigation within the host body still remains puzzling. The high complexity, specificity and diversity of host-recognition strategies suggest that host finding and host recognition are important determinants in the evolution of parasite life cycles. PMID- 16351920 TI - Balancing homeostasis and learning in neural circuits. AB - Neural circuits are remarkably adaptable, providing animals with the ability to modify their behavior on the basis of experience. At the same time, they are extremely robust and maintain stability despite the changes associated with adaptation. This combination of adaptability and stability is difficult to achieve, and it provides a strong constraint on any models of plasticity in neural circuits. New evidence suggests that the effect of action potential timing on synaptic plasticity may be an important element in reconciling homeostasis with adaptability. In particular, spike-timing dependent plasticity can act as both an adaptive and a homeostatic mechanism, controlling overall firing rates and distributions of synaptic efficacies while making neurons selective for certain aspects of their inputs. It can also cause networks that initially represent the present state of a stimulus to predict its future state on the basis of experience, a theoretical result supported by experimental data in behaving rats. PMID- 16351921 TI - Is slowness a learning principle of the visual cortex? AB - Slow feature analysis is an algorithm for extracting slowly varying features from a quickly varying signal. It has been shown in network simulations on one dimensional stimuli that visual invariances to shift and other transformations can be learned in an unsupervised fashion based on slow feature analysis. More recently, we have shown that slow feature analysis applied to image sequences generated from natural images using a range of spatial transformations results in units that share many properties with complex and hypercomplex cells of the primary visual cortex. We find cells responsive to Gabor stimuli with phase invariance, sharpened or widened orientation or frequency tuning, secondary response lobes, end-stopping, and cells selective for direction of motion. These results indicate that slowness may be an important principle of self-organization in the visual cortex. PMID- 16351922 TI - Behaviour as a tool in the assessment of animal welfare. AB - A central issue in animal welfare research is how to assess the welfare state of animals objectively and scientifically. I argue that this issue can be approached by asking two key questions: 1) is the animal physically healthy and 2) does the animal have what it wants? Behaviour is used to answer both of these questions. In the assessment of physical health, it can be used for clinical and pre clinical diagnosis. In the assessment of what animals want, it has a major role through choice and preference testing. It is particularly important that applied ethologists develop methods for assessing welfare in situ--in the places where concern for animal welfare is greatest such as on farms and in zoos. PMID- 16351924 TI - Fractal analysis of narwhal space use patterns. AB - Quantifying animal movement in response to a spatially and temporally heterogeneous environment is critical to understanding the structural and functional landscape influences on population viability. Generalities of landscape structure can easily be extended to the marine environment, as marine predators inhabit a patchy, dynamic system, which influences animal choice and behavior. An innovative use of the fractal measure of complexity, indexing the linearity of movement paths over replicate temporal scales, was applied to satellite tracking data collected from narwhals (Monodon monoceros) (n = 20) in West Greenland and the eastern Canadian high Arctic. Daily movements of individuals were obtained using polar orbiting satellites via the ARGOS data location and collection system. Geographic positions were filtered to obtain a daily good quality position for each whale. The length of total pathway was measured over seven different temporal length scales (step lengths), ranging from one day to one week, and a seasonal mean was calculated. Fractal dimension (D) was significantly different between seasons, highest during summer (D = 1.61, SE 0.04) and winter (D = 1.69, SE 0.06) when whales made convoluted movements in focal areas. Fractal dimension was lowest during fall (D = 1.34, SE 0.03) when whales were migrating south ahead of the forming sea ice. There were no significant effects of size category or sex on fractal dimension by season. The greater linearity of movement during the migration period suggests individuals do not intensively forage on patchy resources until they arrive at summer or winter sites. The highly convoluted movements observed during summer and winter suggest foraging or searching efforts in localized areas. Significant differences between the fractal dimensions on two separate wintering grounds in Baffin Bay suggest differential movement patterns in response to the dynamics of sea ice. PMID- 16351925 TI - The myoseptal system in Chimaera monstrosa: collagenous fiber architecture and its evolution in the gnathostome stem lineage. AB - Recent studies have revealed the 3D morphology and collagen fiber architecture of myosepta in teleostome fishes. Here we present the first data set on the myoseptal structure of a representative of the chondrichthyan clade. We investigate the series of myosepta in the ratfish Chimaera monstrosa (Holocephali) from the anterior to the posterior body using microdissections of cleared and stained specimens, polarized light microscopy of excised myosepta, and histology. The features of the myoseptal system of Chimaera are compared to data from closely related vertebrate groups and are mapped onto a phylogenetic tree to further clarify the characteristics of the myoseptal series in the gnathostome ancestor. The 3D morphology and collagen fiber architecture of the myoseptal series in C. monstrosa resembles that of Teleostomi (Actinopterygii+Sarcopterygii) with regard to several features. Our comparative analysis reveals that some of them have evolved in the gnathostome stem lineage. (1) A series of epineural and epaxial lateral tendons (LTs) along the whole body, and a series of epipleural and hypaxial LTs in the postanal region evolved in the gnathostome stem lineage. (2) The LTs increase in length towards the posterior body (three-fold in Chimaera). Data on Chimaera and some comparative data on actinopterygian fishes indicate that LTs also increase in thickness towards the posterior body, but further data are necessary to test whether this holds true generally. (3) Another conspicuous apomorphic gnathostome feature is represented by multi-layer structures of myosepta. These are formed along the vertebral column by converging medial regions of successive sloping parts of myosepta. (4) The dorsalmost and ventralmost flanking parts of myosepta bear a set of mediolaterally oriented collagen fibers that are present in all gnathostomes but are lacking in outgroups. Preanal hypaxial myosepta are clearly different from epaxial myosepta and postanal hypaxial myosepta in terms of their collagen fiber architecture. In Chimaera, preanal hypaxial myosepta consist of an array of mediolaterally oriented collagen fibers closely resembling the condition in other gnathostome groups and in petromyzontids. Only one series of tendons, the myorhabdoid tendons of the flanking parts of myosepta, have evolved in the stem lineage of Myopterygii (Gnathostomata+Petromyzontida). Similar to LTs, the tendons of this series also increase in length towards the posterior body. In combination with other studies, the present study provides a framework for the design of morphologically based experiments and modeling to further address the function of myosepta and myoseptal tendons in gnathostomes. PMID- 16351926 TI - The role of polyunsaturated fatty acids in the expression of torpor by mammals: a review. AB - Heterothermic mammals increase the proportion of polyunsaturated fatty acids (PUFA) in their body fats prior to entering torpor. Because PUFA have low melting points, it is thought that they play an important role in maintaining the fluidity of depot fats and membrane phospholipids at low body temperatures. However, PUFA are more prone to autoxidation when exposed to reactive oxygen species (ROS) during torpor and during the periodic arousals that characterize hibernation. A lack of PUFA or an excess of PUFA may constrain the use of torpor by heterothermic mammals. We performed a mixed model meta-analysis of 17 controlled-feeding studies to test the effect of dietary PUFA on the depth and expression of torpor by daily heterotherms and hibernators. We also reviewed the literature on the PUFA content of the diet and depot fats of heterothermic mammals to address two principal topics: (1) Do low dietary levels of PUFA reduce the expression of torpor under laboratory conditions and, if so, are free-ranging animals constrained by a lack of PUFA? (2) Do high dietary levels of PUFA result in a reduction in the use, depth, and duration of torpor and, if so, do free ranging animals seek to optimize rather than maximize PUFA intake? Low-PUFA diets consistently increase the lower setpoint for body temperature and minimum metabolic rate for both hibernators and daily heterotherms. Above the lower setpoint, low-PUFA diets usually increase body temperature and metabolic rate and decrease the duration of torpor bouts and this effect is similar for hibernators and daily heterotherms. Free-ranging rodent hibernators have dietary PUFA intakes that are far higher than those of the low-PUFA diets offered in controlled feeding experiments, so these hibernators may never experience the constraints associated with a lack of PUFA. Diets of free-ranging insectivorous bats and echidnas have PUFA levels that are less than half as high as those offered in experimental low-PUFA diets, yet they exhibit deep and extended bouts of torpor. We argue that alternate mechanisms exist for maintaining the fluidity of body fats and that high-PUFA intake may not be a prerequisite for deep and extended bouts of torpor. Four studies indicate that animals that were fed high-PUFA diets are reluctant to enter torpor and show shallower and shorter torpor bouts. Although authors attribute this response to autoxidation, these animals did not have a higher PUFA content in their depot fats than animals where PUFA was shown to enhance torpor. We suggest that these contradictory results indicate inter specific or inter-individual variation in the ability to control ROS and limit autoxidation of PUFA. High dietary levels of PUFA will constrain the expression of torpor only when the oxidative challenge exceeds the capacity of the antioxidant defence system. Studies of diet selection indicate that insectivorous species with low dietary PUFA levels seek to maximize PUFA intake. However, herbivorous species that have access to plants and plant parts of high-PUFA content do not appear to maximize PUFA intake. These data suggest that animals attempt to optimize rather than maximize PUFA intake. The effect of PUFA should be viewed in the light of a cost-benefit trade-off, where the benefit of high PUFA intake is an easier access to low body temperatures and the cost is increased risk of autoxidation. PMID- 16351927 TI - Sialic acids: fascinating sugars in higher animals and man. AB - Sialic acids are acidic monosaccharides, which are among the most important molecules of higher animals, and occur in some microorganisms. They are bound to complex carbohydrates and occupy prominent positions, especially in cell membranes. Their structural diversity is high and, correspondingly, the mechanisms for their biosynthesis are complex. Sialic acid substituents strongly influence the activity of catabolic enzymes, in particular the sialidases, and thus the turnover rate of glycoconjugates. These sugars are involved in manifold cell functions. Due to the surface location of the acidic molecules they shield macromolecules and cells from enzymatic and immunological attacks. But they also represent recognition sites for various physiological receptors as well as for toxins and microorganisms, and thus allow their colonization. Many viruses use sialic acids for the infection of cells. As sialic acids also play a decisive role in tumor biology they prove to be rather versatile molecules that modulate cell biological events in a sensitive way. It is discussed that their evolvement may have stimulated evolution and rendered organisms less vulnerable to environmental attacks. However, disturbance of their metabolism may cause diseases. PMID- 16351928 TI - Neuroanatomical and electrophysiological studies of identified contact chemoreceptors on the ventral ovipositor valve of 3rd instar larvae of lubber grasshoppers (Taeniopoda eques). AB - A large number of contact chemoreceptors are located on the ovipositor valves of adult female grasshoppers. These receptors play an important role in many aspects of grasshopper life such as detecting the chemical composition of the soil before and during oviposition. It is surprising, however, to find these types of receptors on the ovipositor valves of instar larvae which are not able to oviposit. Thus, these receptors may serve functions other than to search for a suitable site for egg laying. Observation under the scanning electron microscope revealed the presence of uniporous basiconic contact chemoreceptors in addition to different types of trichoid mechanoreceptors on the ovipositor valve of lubber grasshopper 3rd instar larvae. Neuroanatomical studies have shown that these sensilla are multiply innervated, containing one mechanosensory neuron and four chemosensory neurons that project locally and intersegmentally. The tip recording technique from single basiconic sensilla demonstrated mechanosensory responses to deflections of the sensillum as well as gustatory activity when in contact with different chemical solutions. The electrophysiological studies have shown that these sensilla serve as contact chemoreceptors and not as olfactory receptors. PMID- 16351929 TI - Basiepidermal nervous system in Nemertoderma westbladi (Nemertodermatida): GYIRFamide immunoreactivity. AB - The Nemertodermatida are a small group of microscopic marine worms. Recent molecular studies have demonstrated that they are likely to be the earliest extant bilaterian animals. What was the nervous system (NS) of a bilaterian ancestor like? In order to answer that question, the NS of Nemertoderma westbladi was investigated by means of indirect immunofluorescence technique and confocal scanning laser microscopy. The antibodies to a flatworm neuropeptide GYIRFamide were used in combination with anti-serotonin antibodies and phalloidin-TRITC staining. The immunostaining revealed an entirely basiepidermal NS. A ring lying outside the body wall musculature at the level of the statocyst forms the only centralisation, the "brain". No stomatogastric NS has been observed. The GYIRFamide immunoreactive part of the "brain" is formed of loosely packed nerve fibres with multiple small neurones and a few large ones. The peptidergic and aminergic patterns of the NS do not correspond to each other: the former is more developed on the ventral side, the latter is more pronounced on the dorsal side. A pair of GYIRFamide immunoreactive nerve cords innervates the ventral side of the animal, the mouth and the male genital opening. The nemertodermatids studied to-date display no common NS pattern. Possible synapomorphies of the Acoelomorpha are discussed. The study demonstrates that the nemertodermatid NS possesses a number of plesiomorphic features and appears more primitive than the NS in other worms, except the Xenoturbellida. The bilaterian ancestor supposedly possessed only a basiepidermal nerve net and had no centralised brain-like structures and no stomatogastric NS. PMID- 16351931 TI - The scapulocoracoid of the Queensland lungfish Neoceratodus forsteri (Dipnoi: Sarcopterygii): morphology, development and evolutionary implications for bony fishes (Osteichthyes). AB - Among bony fishes, the ontogenetic sequence by which the actinopterygian scapulocoracoid develops has been well described, but that of the sarcopterygian scapulocoracoid is poorly known, as the majority of taxa are only known from fossils. To rectify this, the cartilaginous scapulocoracoid of the extant lungfish Neoceratodus forsteri is examined. In initial stages of its development, the scapulocoracoid of Neoceratodus has a simple rounded shape, and supports the glenoid fossa. It appears nearly contemporaneously with the proximal endochondral element (humerus) of the pectoral fin. Pectoral fin elements develop by segmentation from a continuous field of cartilaginous precursor cells extending distally from the glenoid region of the scapulocoracoid. Subsequent scapulocoracoid development produces a ventromedial process, which is not associated with this field of precursor cells. A dorsal process also develops outside this field. Thus, the scapulocoracoid of Neoceratodus may consist of at least two developmentally distinct regions; (1) the ventromedial being homologous with the coracoid of actinopterygians, tetrapods and other jawed vertebrates and (2) a smaller dorsal process, homologous to the scapular region. The two, together with the glenoid region, give an overall triangular shape. The scapulocoracoids of fossil lungfish and other sarcopterygian fishes are also triangular and are composed of scapular and coracoid regions, rather than the 'buttresses' associated with scapulocoracoids of the Actinopterygii and Tetrapoda. PMID- 16351932 TI - Patterns of axial and appendicular movements during aquatic walking in the salamander Siren lacertina. AB - Most studies of salamander locomotion have focused either on swimming or terrestrial walking, but some salamanders also use limb-based locomotion while submerged under water (aquatic walking). In this study we used video motion analysis to describe the aquatic walking gait of Siren lacertina, an elongate salamander with reduced forelimbs and no hindlimbs. We found that S. lacertina uses a bipedal-undulatory gait, which combines alternating use of the forelimbs with a traveling undulatory wave. Each forelimb is in contact with the substrate for about 50% of the stride cycle and forelimbs have little temporal overlap in contact intervals. We quantified the relative timing and frequency of limb and tail movements and found that, unlike the terrestrial gaits of most salamanders, axial and appendicular movements are decoupled during aquatic walking. We found no significant relationship between stride frequency and aquatic walking velocity, but we did find a statistically significant relationship between tailbeat frequency and aquatic walking velocity, which suggests that aquatic walking speed is mainly modulated by axial movements. By comparing axial wavespeed and distance traveled per tailbeat during swimming (forelimbs not used) and aquatic walking (forelimbs used), we found lower wavespeed and greater distance traveled per tailbeat during aquatic walking. These findings suggest that the reduced forelimbs of S. lacertina contribute to forward propulsion during aquatic walking. PMID- 16351933 TI - Absorption and paracellular visualization of fluorescein, a hydrosoluble probe, in intact house sparrows (Passer domesticus). AB - We describe a method to visualize the cellular location of compounds during absorption by the small intestine in intact animals. First, we employed pharmacokinetic methodology to measure the fractional absorption of sodium fluorescein, a small (MW = 376) water-soluble molecule that is widely used as hydrophilic marker molecule for paracellular permeability studies. Based on the hypothesis that the paracellular pathway acts as a sieve, we predicted that fluorescein absorption would be considerable, but less than that of passively absorbed L-glucose which is a smaller molecule (MW = 180). When the two compounds were gavaged into house sparrows simultaneously, the birds absorbed significantly less fluorescein (42 +/- 8%) than L-glucose (82 +/- 7%), as predicted, and absorptions of the two were correlated as one would predict if they shared the same pathway. We removed intestinal tissue 10 min after gavage with sodium fluorescein and determined the cellular location of the compound's fluorescence using confocal laser microscopy. The fluorescent signal was found primarily in the paracellular space. In contrast, in the same type of experiment using instead the similar-sized fluorescent lipophilic compound rhodamine 123 (MW = 381), most fluorescence appeared inside enterocytes, as expected for a compound that diffuses across the apical membrane. Thus, results from all the experiments are consistent with the hypothesis that hydrophilic fluorescein is absorbed primarily via a paracellular pathway. These methods could be applied to visualize absorption pathways of other compounds in other intact animals. PMID- 16351934 TI - Coding of lateral line stimuli in the goldfish midbrain in still and running water. AB - We investigated in goldfish, Carassius auratus, how running water affects the responses of toral lateral line units to a stationary vibrating sphere or to a non-vibrating sphere that moves along the side of the fish. Experiments were conducted in the presence of running water (hydrodynamic noise) to further explore the sensory capabilities of the lateral line with special focus on the morphological sub-modalities. Previous recordings from lateral line nerve fibres in various fish species and the first nucleus of the ascending lateral line pathway in goldfish revealed flow-sensitive and flow-insensitive units. These physiological differences represent, at least in part, the differences in morphology of the lateral line, superficial and canal neuromasts. Following up on these findings we recorded flow-sensitive and flow-insensitive units in the Torus semicircularis of goldfish. In still water, both types of units responded to a vibrating or moving sphere. In running water, neural responses were weaker when the sphere was moved with the flow but were comparable or slightly stronger when the sphere was moved against the flow. In running water, responses of flow sensitive fibres to the vibrating sphere were masked. In contrast, the responses of units insensitive to water flow were not masked. Our data confirm previous findings but also indicate differences when compared to previous reports. We discuss these differences with respect to lateral line morphology, sub-modalities and convergence of different channels of information at higher brain stations. PMID- 16351935 TI - Evolution of ciliary patterns in the Oligotrichida (Ciliophora, Spirotricha) and its taxonomic implications. AB - Although the somatic ciliature of the Oligotrichida typically comprises only a girdle and ventral kinety, a considerable diversity of ciliary patterns occurs. The four main girdle kinety patterns are identically found in tailed and tail less species. The contractile tail has a complicated and unique ultrastructure and is potentially useful for the cell's movement and/or stabilization during feeding. Accordingly, I assume that this structure has evolved only once, namely, in the Tontoniidae nov. fam., and that the different girdle kinety patterns developed convergently in the tailed and tail-less taxa. Further distinct features suggest the establishment of the families Cyrtostrombidiidae nov. fam. (with cyrtos-like pharyngeal fibres and lack of ventral membranelles and endoral) and Pelagostrombidiidae nov. fam. (with neoformation organelle). An attempt is made to reconstruct the evolution of the kinety patterns based on morphologic, ontogenetic, and ultrastructural data. Some genera of tail-less Oligotrichida base on differences in the ciliary pattern; Omegastrombidium nov. gen. is erected for a further girdle kinety pattern. Likewise, the tailed genus Tontonia is split, resulting in two new genera, viz., Pseudotontonia nov. gen. and Spirotontonia nov. gen. Furthermore, the genus Spirostrombidium is split due to the different origin of the parallel course of girdle and ventral kinety, and Parallelostrombidium nov. gen. is established. However, the genus Thigmostrombidium is rejected because its enlarged thigmotactic membranelles are interpreted as an adaptation to the benthic lifestyle, which occurred several times within different girdle kinety patterns. PMID- 16351936 TI - Feeding behavior and kinematics of the lesser electric ray, Narcine brasiliensis (Elasmobranchii: Batoidea). AB - Jaw protrusion is a major functional motif in fish feeding and can occur during mouth opening or closing. This temporal variation impacts the role that jaw protrusion plays in prey apprehension and processing. The lesser electric ray Narcine brasiliensis is a benthic elasmobranch (Batoidea: Torpediniformes) with an extreme and unique method of prey capture. The feeding kinematics of this species were investigated using high-speed videography and pressure transduction. The ray captures its food by protruding its jaws up to 100% of head length (approximately 20% of disc width) beneath the substrate and generating negative oral pressures (< or = 31 kPa) to suck worms into its mouth. Food is further winnowed from ingested sediment by repeated, often asymmetrical protrusions of the jaws (> 70 degrees deviation from the midline) while sand is expelled from the spiracles, gills and mouth. The pronounced ram contribution of capture (jaw protrusion) brings the mouth close enough to the food to allow suction feeding. Due to the anatomical coupling of the jaws, upper jaw protrusion occurs in the expansive phase (unlike most elasmobranchs and similar to bony fishes), and also exhibits a biphasic (slow-open, fast-open) movement similar to tetrapod feeding. The morphological restrictions that permit this unique protrusion mechanism, including coupled jaws and a narrow gape, may increase suction performance, but also likely strongly constrain dietary breadth. PMID- 16351937 TI - How snakes eat snakes: the biomechanical challenges of ophiophagy for the California kingsnake, Lampropeltis getula californiae (Serpentes: Colubridae). AB - In this study we investigated how ophiophagous snakes are able to ingest prey snakes that equal or exceed their own length. We used X-ray video, standard video, dissection, and still X-rays to document the process of ophiophagy in kingsnakes (Lampropeltis getula) feeding on corn snakes (Elaphe guttata). Most kingsnakes readily accepted the prey snakes, subdued them by constriction, and swallowed them head first. In agreement with previous observations of ophiophagy, we found that the predator snake forces the vertebral column of the prey snake to bend into waves. These waves shorten the prey's body axis and allow it to fit inside the gastrointestinal (GI) tract and body cavity of the predator. Dissection of a kingsnake immediately following ingestion revealed extensive longitudinal stretching of the anterior portion of the GI tract (oesophagus and stomach), and no visible incursion of the prey into the intestine. X-ray video of ingestion showed that the primary mechanism of prey transport was the pterygoid walk, with some contribution from concertina-like compression and extension cycles of the predator's vertebral column in two out of three observations. Complete digestion was observed in only one individual, as others regurgitated before digestion was finished. X-ray stills taken every 4 days following ingestion revealed that the corn snakes were about half digested within the first 4 days, and digestion was complete within 15 days. PMID- 16351938 TI - Melano-macrophage centres in the liver of platyfish, Xiphophorus maculatus, Poeciliidae: Teleostei. AB - This paper describes the localization and structure of melano-macrophage centres (MMCs) in the liver of platyfish (Xiphophorus maculatus), compares these centres with corresponding structures in the spleen and kidney of various teleosts and gives suggestions about their functions. The MMCs were closely apposed to the endothelial layer of liver venules covered with pancreas tissue, i.e. portal venules, and contained numerous cells displaying large amounts of Prussian blue precipitations in tissue treated with acid ferrocyanide. Thus, these centres were rich in ferric compounds, probably haemosiderin, and are suggested to be involved in the normal storage, relocation and recycling of iron compounds from effete or damaged red blood cells of the portal venule system in this species. PMID- 16351939 TI - Convergent evolution of the head retraction escape response in elongate fishes and amphibians. AB - Aquatic escape responses have typically been described as C-starts. However, another aquatic escape response, head retraction, occurs in several elongate species, but has never been studied in detail. The goals of this study are to describe the head retraction escape response in a phylogenetically diverse sample of species, to trace the evolution of head retraction in anamniote vertebrates, and to correlate key morphological traits with escape response behavior. In analyzing the evolution of escape behavior, we found that the head retraction escape response has evolved at least six times in anamniote vertebrates. Using independent contrast analysis, the head retraction escape response was found to be correlated with an increase in the total number of vertebrae and an increase in the elongation of an animal. Results from this study indicate a correlation between head retraction as an escape response, elongation of the axial skeleton, and living in structured habitats. PMID- 16351940 TI - Genome sizes and chromosomes in the basal metazoan Hydra. AB - Hydras belong to one of the earliest eumetazoan animal groups, but to date very little is known about their genome sizes, gene numbers, and chromosomes. Here we provide genome size estimates and corresponding karyotypes for five Hydra species. Nuclear DNA contents were assessed by slide-based Feulgen microphotometry. Hydra oligactis possesses the largest genome of 1450 Mbp, followed by similar 1 C capacities in H. carnea (1350 Mbp), H. vulgaris (1250 Mpb) and H. circumcincta (1150 Mbp). The smallest genome of 380 Mbp was determined in H. viridissima. While the number of chromosomes is identical in all five Hydra species (2n = 30), the size of the chromosomes is strictly correlated to the size of the genome, with H. viridissima having conspicuously small chromosomes. The taxonomic and evolutionary significance of the C-value and chromosomal size variation in this ancient group of metazoans as well as its impact on genomic organization and forthcoming genome projects are discussed. PMID- 16351941 TI - The ultrastructure of locust pleuroaxillary "steering" muscles in comparison to other skeletal muscles. AB - The ultrastructure of locust muscles with different function is examined: the pleuroaxillary flight steering muscle is compared with a typical flight (power muscle) and a typical leg muscle, in particular with respect to sarcomere length, tracheation, mitochondria, and sarcoplasmatic reticulum. The pleuroaxillary muscle exhibits some features characteristic of flight muscles but most of the ultrastructure resembles that of leg muscles. This is in agreement with the innervation of this muscle by an octopaminergic neuron, which also innervates leg muscles but no other flight muscles. It also supports the hypothesis that octopaminergic neurons are important metabolic regulators and that the above muscle types exhibit important differences in energy metabolism. PMID- 16351942 TI - Acoustic communication in Okanagana rimosa (Say) (Homoptera: Cicadidae). AB - The cicada Okanagana rimosa (Say) has an acoustic communication system with three types of loud timbal sounds: (i) A calling song lasting several seconds to about 1 min which consists of a sequence of chirps at a repetition rate of 83 chirps per second. Each chirp of about 6 ms duration contains 4-5 pulses. The sound level of the calling song is 87-90 dB SPL at a distance of 15 cm. (ii) An amplitude modulated courtship song with increasing amplitude and repetition rate of chirps and pulses. (iii) A protest squawk with irregular chirp and pulse structure. The spectra of all three types are similar and show main energy peaks at 8-10 kHz. Only males sing, and calling song production is influenced by the songs of other males, resulting in an almost continuous sound in dense populations. In such populations, the calling songs overlap and the temporal structure of individual songs is obscured within the habitat. The calling song of the broadly sympatric, closely related species O. canadensis (Provander) is similar in frequency content, but distinct in the temporal pattern (24 chirps per second, 24 ms chirp duration, eight pulses per chirp) which is likely important for species separation in sympatric populations. The hearing threshold of the auditory nerve is similar for females and males of O. rimosa and most sensitive at 4-5 kHz. Experiments in the field show that female phonotaxis of O. rimosa depends on parameters of the calling song. Most females are attracted to calling song models with a 9 kHz carrier frequency (peak frequency of the calling song), but not to models with a 5 kHz carrier frequency (minimum hearing threshold). Phonotaxis depends on temporal parameters of the conspecific song, especially chirp repetition rate. Calling song production is influenced by environmental factors, and likelihood to sing increases with temperature and brightness of the sky. Correspondingly, females perform phonotaxis most often during sunny conditions with temperatures above 22 degrees C. Non-mated and mated females are attracted by the acoustic signals, and the percentage of mated females performing phonotaxis increases during the season. PMID- 16351944 TI - The nature and significance of invertebrate cartilages revisited: distribution and histology of cartilage and cartilage-like tissues within the Metazoa. AB - Tissues similar to vertebrate cartilage have been described throughout the Metazoa. Often the designation of tissues as cartilage within non-vertebrate lineages is based upon sparse supporting data. To be considered cartilage, a tissue should meet a number of histological criteria that include composition and organization of the extracellular matrix. To re-evaluate the distribution and structural properties of these tissues, we have re-investigated the histological properties of many of these tissues from fresh material, and review the existing literature on invertebrate cartilages. Chondroid connective tissue is common amongst invertebrates, and differs from invertebrate cartilage in the structure and organization of the cells that comprise it. Groups having extensive chondroid connective tissue include brachiopods, polychaetes, and urochordates. Cartilage is found within cephalopod mollusks, chelicerate arthropods and sabellid polychaetes. Skeletal tissues found within enteropneust hemichordates are unique in that the extracellular matrix shares many properties with vertebrate cartilage, yet these tissues are completely acellular. The possibility that this tissue may represent a new category of cartilage, acellular cartilage, is discussed. Immunoreactivity of some invertebrate cartilages with antibodies that recognize molecules specific to vertebrate bone suggests an intermediate phenotype between vertebrate cartilage and bone. Although cartilage is found within a number of invertebrate lineages, we find that not all tissues previously reported to be cartilage have the appropriate properties to merit their distinction as cartilage. PMID- 16351945 TI - Formation of the corneous layer in the epidermis of the tuatara (Sphenodon punctatus, Sphenodontida, Lepidosauria, Reptilia). AB - The formation of the stratum corneum in the epidermis of the reptile Sphenodon punctatus has been studied by histochemical, immunohistochemical, and ultrastructural methods. Sulfhydryl groups are present in the mesos and pre-alpha layer but disappear in the keratinized beta-layer and in most of the mature alpha layer. This suggests a complete cross-linking of keratin filaments. Tyrosine increases in keratinized layers, especially in the beta-layer. Arginine is present in living epidermal layers, in the presumptive alpha-layer, but decreases in keratinized layers. Histidine is present in corneous layers, especially in the intermediate region between the alpha- and a new beta-layer, but disappears in living layers. It is unknown whether histidine-rich proteins are produced in the intermediate region. Small keratohyalin-like granules are incorporated in the intermediate region. The plane of shedding, as confirmed from the study on molts, is located along the basalmost part of the alpha-layer and may involve the degradation of whole cells or cell junctions of the intermediate region. A specific shedding complex, like that of lizards and snakes, is not formed in tuatara epidermis. AE1-, AE2-, or AE3-positive alpha-keratins are present in different epidermal layers with a pattern similar to that previously described in reptiles. The AE1 antibody stains the basal and, less intensely, the first suprabasal layers. Pre-keratinized, alpha- and beta-layers, and the intermediate region remain unlabeled. The AE2 antibody stains suprabasal and forming alpha- and beta-layers, but does not stain the basal and suprabasal layers. In the mature beta-layer the immunostaining disappears. The AE3 antibody stains all epidermal layers but disappears in alpha- and beta-layers. Immunolocalization for chick scale beta-keratins labels the forming and mature beta-layer, but disappears in the mesos and alpha-layer. This suggests the presence of common epitopes in avian and reptilian beta-keratins. Low molecular weight alpha keratins present in the basal layer are probably replaced by keratins of higher molecular weight in keratinizing layers (AE2-positive). This keratin pattern was probably established since the beginning of land adaptation in amniotes. PMID- 16351946 TI - Extraembryonic membrane development in a reproductively bimodal lizard, Lacerta (Zootoca) vivipara. AB - Reproductive mode has been remarkably labile among squamate reptiles and the evolutionary transition from oviparity to viviparity commonly has been accompanied by a shift in the pattern of embryonic nutrition. Structural specializations for placental transfer of nutrients during intrauterine gestation are highly diverse and many features of the extraembryonic membranes of viviparous species differ markedly from those of oviparous species. However, because of a high degree of evolutionary divergence between the species used for comparisons it is likely that the observed differences arose secondarily to the evolution of viviparity. We studied development of the extraembryonic membranes and placentation in the reproductively bimodal lizard Lacerta vivipara because the influence of reproductive mode on the structural/functional relationship between mothers and embryos can best be understood by studying the most recent evolutionary events. Lecithotrophic viviparity has evolved recently within this species and, although populations with different reproductive modes are allopatric, oviparous and viviparous forms interbreed in the laboratory and share many life history characteristics. In contrast to prior comparisons between oviparous and viviparous species, we found no differences in ontogeny or structure of the extraembryonic membranes between populations with different reproductive modes within L. vivipara. However, we did confirm conclusions from previous studies that the tertiary envelope of the egg, the eggshell, is much reduced in the viviparous population. These conclusions support a widely accepted model for the evolution of squamate placentation. We also found support for work published nearly 80 years ago that the pattern of development of the yolk sac of L. vivipara is unusual and that a function of a unique structure of squamate development, the yolk cleft, is hematopoiesis. The structure of the yolk sac splanchnopleure of L. vivipara is inconsistent with a commonly accepted model for amniote yolk sac function and we suggest that a long standing hypothesis that cells from the yolk cleft participate in yolk digestion requires further study. PMID- 16351947 TI - Nutritional development of feeding strategies in arctic ruminants: digestive morphometry of reindeer, Rangifer tarandus, and muskoxen, Ovibos moschatus. AB - Reindeer have been classified as intermediate feeders and muskoxen as grazers based on differences in digestive morphology and consumption of fibrous plants. We hypothesized that the digestive morphology of young (< 2 months) reindeer and muskoxen anticipates transitions in diet and determines the feeding strategy of each species at adulthood. We compared structural morphology and rates of cell division in the rumen, abomasum, duodenum and liver of reindeer and muskoxen as neonates (1 day old), during the transition from milk to forage (30-60 days old) and in adults (> 7 yr). Development in utero provided the neonate with a functioning mucosa of the gastric abomasum and duodenal mucosa with high surface enlargement for digestion and absorption of concentrated milks. Transition to forage was preceded by changes in ruminal papillae structure that increased surface area and likely contributed to active fermentation by 60 days of age. The abomasum also increased in acid-secreting parietal cells during the transition to forage, which may enhance digestion of plant and microbial proteins. Rates of cell division also indicated a sustained differentiation of tissue structure during the transitional period. Young arctic ruminants expressed digestive structures that preceded full function, which indicated the ultimate feeding strategy of each species. For example, the rumen of young muskoxen had thick cornified epithelia and muscle layers that would provide ruminal mucosa with better protection from fibrous abrasion and enhance motility of bulky diets. Conversely, young reindeer had more complex papillary shapes in the rumen and more foliate villi in the duodenum, indicating a greater absorptive capacity of these structures than in muskoxen. Ontogenetic programs, therefore, play the primary role for digestive development of reindeer and muskoxen and determine the nutritional strategies of adults. PMID- 16351948 TI - Motor pattern control for increasing crushing force in the striped burrfish (Chilomycterus schoepfi). AB - The relationship between muscular force modulation and the underlying nervous system control signals has been difficult to quantify for in vivo animal systems. Our goal was to understand how animals alter muscle activation patterns to increase bite forces and to evaluate how accurate these patterns are in predicting crushing forces. We examined the relationship between commonly used measures of cranial muscle activity and force production during feeding events of the striped burrfish (Chilomycterus schoepfi), a mollusc crushing specialist. We quantified the force required to crush a common gastropod prey item (Littorina irrorata) of burrfish using a materials testing device. Burrfish were fed these calibrated prey items while we recorded electromyograms (EMGs) from the main jaw closing muscles (adductor mandibulae A1beta, A2alpha, and A2beta). We quantified EMG activity by measuring the burst duration, rectified integrated area, and then calculated the intensity of activity from these two variables. Least squares regressions relating force to crush (Fcrush) and all EMG variables were calculated for each fish. Multiple regression analyses were used to determine how much of the variation in Fcrush could be explained by muscle activation patterns. We found that 20 cm burrfish are capable of generating extremely high crushing forces (380 N peak force) primarily by increasing the duration of muscle activity. EMG variables explained 71% of the total variation in force production. After accounting for the inherent variation in Fcrush of snails, EMGs do a very good job of predicting bite forces for these fish. PMID- 16351950 TI - The mole's thumb -- evolution of the hand skeleton in talpids (Mammalia). AB - Osteological specimens representing 15 out of the 16 currently recognized talpid genera were examined and scored for seven discrete morphological characters of the hand. The phylogenetic distribution of these characters was studied in the context of alternative hypotheses of talpid relationships, using three species of shrews and a hedgehog as outgroups. All talpids show a similar number and arrangement of carpal bones. The most obvious differences concern the presence of additional sesamoid bones, the relative size of the os falciforme when present, and the degree of fusion of the scaphoid and lunate in the proximal carpal row. Marked differences in the relative length and proportions of the metacarpals also exist. The development of the carpals in Talpa europaea was studied through examination of histological sections of the hand of an embryo and a neonate. Whereas carpal anatomy in the neonate mirrors the arrangement and proportions of the adult, in the embryo the scaphoid and lunate are still separate, there are no signs of the os falciforme, and the size proportions of metacarpals to carpals are obviously different to those of the adult. A prehallux or tibial sesamoid, serial homologue to the os falciforme or prepollex (a radial sesamoid), does not have an obvious functional role, and its presence might be the result of a common epigenetic control in the hand and the foot resulting in a non-adaptive structure in the latter. PMID- 16351951 TI - Cephalic neural crest cells and the evolution of craniofacial structures in vertebrates: morphological and embryological significance of the premandibular mandibular boundary. AB - The vertebrate head characteristically has two types of mesenchyme: the neural crest-derived ectomesenchyme and the mesoderm derived mesenchyme. Conserved patterns of development in various animal taxa imply the presence of shared inductive events for cephalic mesenchyme. These developmental programs can serve as developmental constraints that emerge as morphological homology of embryonic patterns. To understand the evolutionary changes in the developmental programs that shape the skull, we need to separate ancestral and derived patterns of vertebrate craniogenesis. This review deals with the terminology for neural crest cell subpopulations at each developmental stage, based on the topographical relationships and possible mechanisms for specification. The aim is to identify the changes that could have occurred in the evolutionary history of vertebrates. From comparisons of a lamprey species, Lethenteron japonicum, with gnathostomes it is clear that the initial distribution of cephalic crest cells is identical in the two animal lineages. In all vertebrate embryos, the trigeminal crest (TC) cells of an early pharyngula are subdivided into three subpopulations. At this stage, only the posterior subpopulation of the TC cells is specified as the mandibular arch, as compared to the more rostral components, the 'premandibular crest cells'. Later in development, the local specification patterns of the lamprey and the gnathostomes differ, so that homology cannot be established in the craniofacial primordia, including the oral apparatus. Therefore, embryological terminology should reflect these hierarchical patterns in developmental stages and phylogeny. PMID- 16351952 TI - Resurrection of Bohadschia bivittata from B. marmorata (Holothuroidea: Holothuriidae) based on behavioral, morphological, and mitochondrial DNA evidence. AB - Behavior, color, body size, spicules, and mitochondrial DNA were examined in two morphs from the Bohadschia marmorata (Jaeger, 1833) species complex in Micronesia to test whether they are conspecific. This complex consists of eight morphs that have been described as separate species and combined in various ways for over a century. We examined the classic B. marmorata type and the type originally described as the species B. bivittata (Mitsukuri, 1912); B. bivittata was combined with B. marmorata by Panning (1944). Several observations and a phylogenetic analysis led us to conclude that B. marmorata and B. bivittata should return to their status as separate species. First, B. marmorata lives in shallow areas with strong currents, and B. bivittata lives on open sand between corals in deeper water. Second, the coloration of B. bivittata is distinct from B. marmorata, and although specimens collected on Yap Island differed from Mitsukuri's original description of B. bivittata, no specimens were collected with coloration intermediate between B. bivittata and B. marmorata. Third, spicules are more highly branched, perforated, and spiked in B. bivittata than in B. marmorata (and, in our study, spicule complexity did not correlate with body size). Finally, our phylogenetic analysis, based on partial nucleotide sequences of 16s, 12s, and COI mitochondrial genes, resulted in a tree--(Pearsonothuria graeffei (Bohadschia marmorata) (B. argus (B. bivittata)))--which shows that B. marmorata and B. bivittata are not even sister species, with B. bivittata more closely related to B. argus. Support for the clades for each Bohadschia species was strong, but the clade containing B. argus and B. bivittata had weaker support. Color and spicule examinations made of preserved B. marmorata-complex specimens from the Indo-Pacific as well as behavioral observations in the field also support the resurrection of B. bivittata. PMID- 16351953 TI - Gumfooted lines in black widow cobwebs and the mechanical properties of spider capture silk. AB - Orb-weaving spiders produce webs using two types of silk that have radically different mechanical properties. The dragline silk used to construct the supporting frame and radii of the web is stiff and as strong as steel, while the capture spiral is much weaker but more than ten times as extensible. This remarkable divergence in mechanical properties has been attributed to the aqueous glue that coats the capture spiral, which is thought to decrease capture spiral stiffness and increase its extensibility. However, discerning the effect of the aqueous glue on fiber performance is complicated because dragline silk and the capture spiral are assembled from different proteins, which may also affect mechanical performance. Here, we use the sticky gumfooted lines of black widow cobwebs to test the effect of the addition of aqueous glue on the mechanical properties of dragline silk. We also surveyed orb-webs spun by a broad range of species for bundles of looped silk. Such bundles, termed windlasses, have been thought to increase capture spiral extensibility by "paying out" additional lengths of silk. Our results suggest that neither plasticization of silk by aqueous glue nor excess silk in windlasses can by themselves account for the remarkable extensibility of orb-weaver capture silk compared to other spider silks. This argues that the unique amino acid motifs of the flagelliform fibroins that constitute the core of the capture spiral play an essential role in capture silk's extreme extensibility. PMID- 16351954 TI - Duplicated Pax6 genes in Glomeris marginata (Myriapoda: Diplopoda), an arthropod with simple lateral eyes. AB - Composite (facetted) eyes comprised by several units, termed ommatidia, are an ancestral feature in the arthropods. Some arthropods, however, do not possess composite eyes, obviously by secondary reduction. Reductions on the level of conserved eye developmental genes are one possibility to reduce the visual system. The genes of the Pax6 family have been shown to be key regulators of visual system development in a wide variety of animals. Reduction of Pax6 expression may therefore be expected in a species with reduced eyes. Here I have investigated the myriapod Glomeris marginata that displays very simple eyes. Glomeris, however, possesses two Pax6 genes that, based on their sequence, are similar to Drosophila eyeless (ey) and twin of eyeless (toy), respectively. Both genes are highly expressed in the optic lobes and the ventral nerve cord of developing embryos. Furthermore, homologs of other high-ranking eye developmental genes like hedgehog, decapentaplegic, dachshund, and homothorax are expressed in the optic lobes. This indicates that eye reduction in Glomeris is not realized at the level of the Pax6 genes or other genes on the upper levels of the eye development network. I suggest instead that the simple eyes of Glomeris are the product of changes at a much lower level in the network, probably at the level of genes directly regulating ommatidia development or ommatidia number and arrangement. PMID- 16351955 TI - The effect of larval age on developmental changes in the polyp prepattern of a hydrozoan planula. AB - The larvae of many marine organisms including hydrozoans are lecithotrophic and will not feed until after metamorphosis. In hydrozoans the aboral region of the planula becomes the holdfast and stolon, while the oral region becomes the stalk and hydranth that grows out of the holdfast following metamorphosis. If metamorphosis is delayed, the portion of the planula allocated to form holdfast and stolon shrinks and the region that forms the hydranth increases in size. Planulae also have the ability to regenerate their polyp prepattern. When the aboral region of the planula that does not normally form a hydranth is isolated and metamorphosis is delayed, it acquires the capacity to form a hydranth from the holdfast. A relatively high proportion of entodermal cells of young planulae engage in DNA synthesis (BrdU labeling index); as planulae age, the labeling index falls close to zero. When the polyp prepattern is modified during planula regeneration, entodermal cells are induced to engage in DNA synthesis. If DNA synthesis is inhibited in planulae, the polyp prepattern changes during regeneration and age-related developmental changes in planula are inhibited, suggesting that DNA synthesis is a necessary part of the pattern respecification process. PMID- 16351957 TI - A comparative study of embryonic development of some bird species with different patterns of postnatal growth. AB - Some studies show that birds with high postnatal growth rates (e.g. altricial species) are characterized by a rapid early development of "supply" organs, such as digestive organs. Birds with low postnatal growth rates (e.g. precocial species) exhibit a slower early development of these organs and a more rapid early development of other "demand" organs, such as brain, muscles, skeleton and feathers. To test whether these differences can be traced back to early embryonic development and whether they can be associated with changes in developmental timing, i.e. heterochrony, we compared embryos of the precocial quail and the altricial fieldfare, two bird species with low and high postnatal growth rates, respectively. We used classical staging techniques that use developmental landmarks to categorize embryonic maturity as well as morphological measurements. These techniques were combined with immune detection of muscle specific proteins in the somites. Our data showed that the anlagen of the head, brain and eyes develop earlier in the quail than in the fieldfare in contrast to the gut which develops earlier in the fieldfare than in the quail. Our data also showed that the quail and the fieldfare displayed different rates of myotome formation in the somites which contribute to muscle formation in the limbs and thorax. We believe these observations are connected with important differences in neonatal characteristics, such as the size of the brain, eyes, organs for locomotion and digestion. This leads us to the conclusion that selection for late ontogenetic characteristics can alter early embryonic development and that growth rate is of fundamental importance for the patterning of avian embryonic development. It also appears that this comparative system offers excellent opportunities to test hypotheses about heterochrony. PMID- 16351958 TI - Flexible remodeling of organ size during spring migration of the garden warbler (Sylvia borin). AB - The energetic demands of long-distance migratory birds change drastically, depending on the stage of their life cycle. Changing demands are reflected in the up and down regulation of adipose tissue and organ mass. This paper presents new data on organ size changes during different stages of spring migration of garden warblers (Sylvia borin). Phenotypic mass changes were quantified in 13 organs of birds caught in Tanzania, Ethiopia and Egypt. We also sampled birds after a simulated stopover in Egypt. Some organs increased in mass up to about 1.5-fold during migration from Tanzania to Ethiopia, while some remained unchanged or even decreased in mass. During flight across the Sahara, nearly all organ masses including heart and flight muscles were reduced. Exceptionally large reductions (approximately 50%) were observed for liver, bile, spleen, kidney and digestive tract organs. The only exceptions were the testes, which increased 4-fold in mass. During the simulated stopover in Egypt, a significant recovery was observed for kidney, liver, heart, proventriculus, and small intestine. The testes continued to increase in mass. Flexible remodeling of organ size in the course of spring migration thus comprises significant changes for all quantified organs, with a variety of organ-specific patterns. Individual organ patterns are differentially shaped by functional aspects according to the different organ requirements in the alternation of flight and stopover phases, energetics, future demands, and protein requirements. Anticipatory mechanisms account for the size change of the testes, and we suggest the same for the kidney and the gall bladder. PMID- 16351959 TI - The behavioral responses of amphibians and reptiles to microgravity on parabolic flights. AB - In the present study, we exposed 53 animals from 23 different species of amphibians and reptiles to microgravity (mug). This nearly doubles the number of amphibians and reptiles observed so far in mug. The animals were flown on a parabolic flight, which provided 20-25s of mug, to better characterize behavioral reactions to abrupt exposure to mug. Highly fossorial limbless caecilians and amphisbaenians showed relatively limited movement in mug. Limbed quadrupedal reptiles that were non-arboreal in the genera Leiocephalus, Anolis, and Scincella showed the typical righting response and enormous amounts of body motion and tail rotation, which we interpreted as both righting responses and futile actions to grasp the substrate. Both arboreal and non-arboreal geckos in the genera Uroplatus, Palmatogecko, Stenodactylus, Tarentola, and Eublepharis instead showed a skydiving posture previously reported for highly arboreal anurans. Some snakes, in the genera Thamnophis and Elaphe, which typically thrashed and rolled in mug, managed to knot their own bodies with their tails and immediately became quiescent. This suggests that these reptiles gave stable physical contact, which would indicate that they were not falling, primacy over vestibular input that indicated that they were in freefall. The fact that they became quiet upon self embrace further suggests a failure to distinguish self from non-self. The patterns of behavior seen in amphibians and reptiles in mug can be explained in light of their normal ecology and taxonomic relations. PMID- 16351960 TI - Relationship between diet and helminths in Gallotia caesaris (Sauria: Lacertidae). AB - Diet and helminth fauna were analysed in Gallotia caesaris, a small lacertid lizard endemic to El Hierro (Canary Islands, Spain) in order to study the evolutionary and functional relationships between the two traits. This species is omnivorous but consumed a high proportion (82.13%) of plant matter including not only seeds but also leaves and other vegetative parts. Helminth fauna included many helminth species typical of herbivorous reptiles. Both herbivory and helminth presence were higher than expected for a lizard of its size. Comparison with other lacertids suggests that both traits result from an adaptation to insular conditions but that some "evolutionary time" to develop them is needed. Canarian Gallotia lizards, a separate lineage evolving for a long time in insularity, constitutes the most advanced lacertid group in this way. Nevertheless, results for G. caesaris indicate that helminth fauna also changes seasonally tracking variation in diet (and herbivory) throughout the year, which suggests a dynamic interaction between diet and helminth parasites. PMID- 16351961 TI - Delayed embryonic development in the Indian short-nosed fruit bat, Cynopterus sphinx. AB - The unusual feature of the breeding cycle of Cynopterus sphinx at Varanasi is the significant variation in gestation length of the two successive pregnancies of the year. The aim of this study was to investigate whether the prolongation of the first pregnancy in C. sphinx is due to delayed embryonic development. The first (winter) pregnancy commences in late October and lasts until late March and has a gestation period of about 150 days. The second (summer) pregnancy commences in April and lasts until the end of July or early August with a gestation period of about 125 days. Changes in the size and weight of uterine cornua during the two successive pregnancies suggest retarded embryonic growth during November and December. Histological analysis during the period of retarded embryonic development in November and December showed a slow gastrulation process. The process of amniogenesis was particularly slow. When the embryos attained the early primitive streak stage, their developmental rate suddenly increased considerably. During the summer pregnancy, on the other hand, the process of gastrulation was much faster and proceeded quickly. A comparison of the pattern of embryonic development for 4 consecutive years consistently showed retarded or delayed embryonic development during November and December. The time of parturition and post-partum oestrus showed only a limited variation from 1 year to another. This suggests that delayed embryonic development in C. sphinx may function to synchronize parturition among females. The period of delayed embryonic development in this species clearly coincides with the period of fat deposition. The significance of this correlation warrants further investigation. PMID- 16351962 TI - Cranial movements during suction feeding in teleost fishes: Are they modified to enhance suction production? AB - Suction is produced during prey capture by most teleost fishes. Here, we ask two questions about the functional basis of suction feeding. First, is there variation in the kinematic pattern produced by different species while suction feeding? Second, do species termed 'suction specialists' demonstrate similar modifications to their feeding behavior? We used 10 kinematic variables in a principal component analysis to identify axes of variation among 14 suction feeding teleost species (representing nine families and five orders within the Euteleostei) that demonstrate different feeding habits and habitats. MANOVA and Tukey post hoc tests were used to assess differences among species. Most species clustered together on the principal component axes, suggesting a generalized mechanism that facilitates unidirectional flow. Typically, only one species stood out as 'extreme' on each functional axis, and a species that stood out on one axis did not stand out on others. Only one species, the flatfish Pleuronichthys verticalis, an obligate benthic feeder, demonstrated modifications consistent with enhanced suction production. This species displayed a suite of changes that should enhance suction production, including large hyoid depression, large cranial rotation, and small gape. We suggest that suction performance may be greatest in such obligate benthic feeders because cranial morphology is highly modified and prey are captured from the substrate. PMID- 16351963 TI - The ontogeny of feeding kinematics in a giant salamander Cryptobranchus alleganiensis: Does current function or phylogenetic relatedness predict the scaling patterns of movement? AB - Studies of the scaling of feeding movements in vertebrates have included three species that display both near-geometric growth and isometry of kinematic variables. These scaling characteristics allow one to examine the "pure" relationship of growth and movement. Despite similar growth patterns, the feeding movements of toads (Bufo) slow down more with increasing body size than those of bass (Micropterus), and sharks (Ginglymostoma). This variation might be due to major differences in the mechanism of prey capture; the bass and sharks use suction to capture prey in water, while the toad uses tongue prehension to capture prey on land. To investigate whether or not these different scaling patterns are correlated with differences in feeding mechanics, we examined the ontogenetic scaling of prey capture movements in the hellbender salamander (Cryptobranchus alleganiensis), which also has near-geometric growth. The hellbender suction feeds in the same general manner as the teleosts and shark, but is much more closely related to the toad. The feeding movements of the hellbender scale more similarly to the feeding movements of toads than to those of fishes or sharks, indicating that phylogenetic relatedness rather than biomechanical similarity predicts ontogenetic scaling patterns of movement. PMID- 16351964 TI - Phylogeny of three parapatric species of desert ants, Cataglyphis bicolor, C. viatica, and C. savignyi: a comparison of mitochondrial DNA, nuclear DNA, and morphological data. AB - Due to morphological comparisons the Tunisian desert ant species Cataglyphis bicolor has been divided into three parapatric species: C. bicolor, C. viatica, and C. savignyi. The species status of the latter is supported by sequence analyses of the mitochondrial CO1 and CO2 region, while analyses of the same mitochondrial region lacked resolution for the separation of C. bicolor and C. viatica. However, the geographic distribution of mtDNA haplotypes points to different population viscosities with C. bicolor queens having longer migration distances than queens of C. viatica. Furthermore, by the use of microsatellites we excluded ongoing gene flow between geographically overlapping populations of C. bicolor and C. viatica, and hence support the morphology-based three-species hypothesis. Concerning the ongoing discussion on the future roles of morphology and molecular biology in systematics we call for a combination of both whenever possible. PMID- 16351966 TI - Pigmentation development, defects, and patterning in summer flounder (Paralichthys dentatus). AB - Flounders offer unique opportunities to study the cytological basis of vertebrate pigmentation. Individual skin pigment cells are clearly visible at hatching, and flounder ontogeny includes a dramatic shift in overall pigmentation (from symmetrical to asymmetrical) during metamorphosis. Moreover, several types of malpigmentation occur in hatchery populations; although much effort has gone into reducing the frequency of such defects, their etiology remains poorly understood, and they have rarely been described at the cellular level. In this paper, we use light and fluorescence microscopy to describe the cytological basis of normal developmental changes and of common types of malpigmentation. We then discuss the implications of these observations for underlying patterning mechanisms. PMID- 16351967 TI - Sea snakes (Lapemis curtus) are sensitive to low-amplitude water motions. AB - The sea snake Lapemis curtus is a piscivorous predator that hunts at dusk. Like land snakes, sea snakes have scale sensillae that may be mechanoreceptive, i.e. that may be useful for the detection of water motions produced by prey fish. In addition, inner ear hair cells of sea snakes may also be involved in the detection of hydrodynamic stimuli. We generated water motions and pressure fluctuations with a vibrating sphere. In the test range 50-200 Hz evoked potentials were recorded from the midbrain of L. curtus in response to vibrating sphere stimuli. In terms of water displacement the lowest threshold amplitudes were in the frequency range 100-150 Hz. In this range peak-to-peak water displacement amplitudes of 1.8 microm (at 100 Hz) and 2.0 microm (150 Hz) generated a neural response in the most sensitive animal. Although this low sensitivity may be sufficient for the detection of fish-generated water motions, it makes it unlikely that L. curtus has a special hydrodynamic sense. PMID- 16351968 TI - Correlating diet and digestive tract specialization: examples from the lizard family Liolaemidae. AB - A range of digestive tract specializations were compared among dietary categories in the family Liolaemidae to test the hypothesis that herbivores require greater gut complexity to process plant matter. Additionally, the hypothesis that herbivory favors the evolution of larger body size was tested. Lastly, the association between diet and hindgut nematodes was explored. Herbivorous liolaemids were larger relative to omnivorous and insectivorous congeners and consequently had larger guts. In addition, small intestine length of herbivorous liolaemids was disproportionately longer than that of congeners. Significant interaction effects between diet and body size among organ dimensions indicate that increases in organ size occur to a greater extent in herbivores than other diet categories. For species with plant matter in their guts, there was a significant positive correlation between the percentage of plant matter consumed and small intestine length. Herbivorous liolaemids examined in this study lacked the gross morphological specializations (cecum and colonic valves) found in herbivores in the families Iguanidae and Agamidae. A significantly greater percentage of herbivorous species had nematodes in their gut. Of the species with nematodes, over 95% of herbivores had nematodes only in the hindgut. Prevalence of nematodes in the hindgut of herbivores was 2 x that of omnivores and 4 x that of insectivores. PMID- 16351969 TI - How can nematodes mate without spicules? Function of the male gonoduct glands in the roundworm Myolaimus. AB - Males of roundworms (Nematoda) usually possess cuticular copulatory organs (spicules) that are inserted in the female's vulva to attach the male to the female and to widen the vulva against the inner body pressure for sperm transfer. Among free-living nematodes, the only exception of this rule is Myolaimus where the males lack spicules. Until now there exist no reports on how mating is achieved in Myolaimus. Here we show that sperm transfer in Myolaimus apparently involves at least six different secretions of the male gonoduct that are pumped into a sack-like cuticular protrusion of the female's vulva to form a spermatophore-like capsule. The role of gonoduct glands in male nematodes (even in the model organism Caenorhabditis elegans) is poorly understood. Here we present the first study explaining the role of different vas deferens gland products in nematodes and argue that Myolaimus males lost their spicules as a result of sperm competition. PMID- 16351970 TI - Neuroanatomical and immunocytochemical studies of the head retractor muscle innervation in the pond snail, Lymnaea stagnalis L. AB - Axonal tracing and immunocytochemical techniques were used to study the innervation of the head retractor muscle (HRM) in the pond snail Lymnaea stagnalis L. Fibers of both the superior and inferior cervical nerves which innervate the HRM form endings that comply with the structure of chemical synapses. The somata of neurons with axons in these nerves are located in all except the buccal ganglia of the central nervous system, and this seems to be a special feature of the HRM motor system. By staining the filamentous actin with Oregon-green conjugated phalloidin, we demonstrated that the HRM has a multiterminal innervation and one muscle fiber can contain several synaptic endings which appear to be both morphologically and physiologically different. The morphological diversity of synaptic vesicles suggests a multiplicity of neurotransmitters acting on these nerve-muscle junctions. Immunocytochemical evidence was found for a strong serotonergic and FMRFamidergic innervation of muscle fibers through axons of the inferior cervical nerve. The thin fibers of the inferior cervical nerve possess immunoreactivity to glutamate, gamma aminobutyric acid (GABA) and choline-acetyltransferase, and form sparser innervation patterns in the muscle. Our results indicate that several neurotransmitters are present in the nerves innervating the Lymnaea HRM and may therefore participate in the control of this muscle. The possible behavioral significance of such different neurotransmitter sets involved in the regulation of contractions is discussed. PMID- 16351971 TI - Phonotaxis of the female parasitoid Emblemasoma auditrix (Diptera, Sarcophagidae) in relation to number of larvae and age. AB - The dipteran parasitoid Emblemasoma auditrix locates its host acoustically. Analysis showed that phonotactic female flies usually carry fully developed larvae within their uteri. The mean number of larvae per female at the beginning of the season was 37.9 (range from 10 to 50). The number of larvae decreased rapidly with increasing singing activity of the host cicada (Okanagana rimosa). In high-density host populations the parasitoid is likely to become egg-limited. A possible selective phonotactic responsiveness depending on the number of larvae or the age of the female was tested with song models. Phonotaxis depended on both the temporal structure and the frequency content, but in the field no correlation was found between the number of larvae and the preferences for the acoustic signal. Experiments in the laboratory showed that flies without host contact broadened their phonotactic stimulus range with age. PMID- 16351974 TI - Integrative organismal biology: papers in honor of Professor Marvalee H. Wake. PMID- 16351972 TI - The pink-blue spot syndrome in Acropora eurystoma (Eilat, Red Sea): a possible marker of stress? AB - The appearance of pink-blue spots (termed here as pink-blue spot syndrome - PBSS) in the branching coral Acropora eurystoma from the Gulf of Eilat, Red Sea, is described. We monitored 18 transects (10 x 1 m2 each) in front of the H. Steinitz Marine Laboratory (Eilat), at 3, 6 and 9 m depth, during March and August in 2001 and 2002. Transect measurements revealed high frequencies of colonies with PBSS (up to 100% of colonies) between 3 and 9 m depth. Ten PBSS-affected colonies of A. eurystoma were labelled and monitored for the development of spots. From March to August 2001, the number of spots per colony increased and remained constantly high at both sampling dates in 2002. Spot size ranged between 7 and 149 mm2. Spots were primarily recorded in areas where coral tissues contacted foreign biological matter, either around regenerative wounds or when surrounded by encrusting organisms, in fast-growing areas and in allogeneic interactions. A preliminary biochemical examination suggested that the pink-blue pigment in A. eurystoma is part of a family of compounds (pocilloporin) responsible for the pink-blue colours in pocilloporid and acroporid corals. Pink-blue colour could be experimentally induced in A. eurystoma by tissue-to-tissue contacts between distressed and non-distressed allogeneic branches. PBSS was also induced in healthy coral tissue by contact with inert objects, e.g., by bandaging a branch with plastic strips. Any specific pink-blue colour spots faded within 1-3 months from onset. These results suggest that PBSS in A. eurystoma may not be considered a regular coral disease, but rather a locally induced syndrome caused by restricted environmental and/or biological stress conditions. PMID- 16351975 TI - TRIBUTE: In Goethe's Wake: Marvalee Wake's conceptual contributions to the development and evolution of a science of morphology. AB - De-crying the typological approach in much of the teaching of morphology, from the outset of her career Marvalee Wake advocated a synthetic, mechanistic and pluralistic developmental and evolutionary morphology. In this short essay, I do not evaluate Wake's contributions to our knowledge of the morphology of caecilians, nor her contributions to viviparity, both of which are seminal and substantive, nor do I examine her role as mentor, supervisor and collaborator, but assess her broader conceptual contributions to the development and evolution of morphology as a science. One of the earliest morphologists to take on board the concept of constraint, she viewed constraint explicitly in relation to adaptation and diversity. Her approach to morphology as a science was hierarchical - measure form and function in a phylogenetic context; seek explanations at developmental, functional, ecological, evolutionary levels of the biological hierarchy; integrate those explanations to the other levels. The explanatory power of morphology thus practised allows morphology to inform evolutionary biology and evolutionary theory, and paves the way for the integrative biology Wake has long championed. PMID- 16351976 TI - Androgynous rex - the utility of chevrons for determining the sex of crocodilians and non-avian dinosaurs. AB - The sex of non-avian dinosaurs has been inferred on numerous occasions using a variety of anatomical criteria, but the efficacy of none has been proven. Nearly 50 years ago Romer suggested that the cranial-most or first chevron in the tails of some reptiles, including crocodilians, is sexually dimorphic. Recent work on this subject purportedly substantiated that the female first chevron articulates in a more caudal position than in males. Furthermore, it was concluded that this element is shorter in females. These phenotypic attributes theoretically provide a broader cloacal passageway for eggs by ovipositing females and a greater attachment area for male "penile retractor muscles". Because theropod dinosaurs such as Tyrannosaurus rex presumably show similar variation in chevron anatomy, the same criteria has been advocated for sexing dinosaurs. We tested the neontological model for the chevron sexual dimorphism hypothesis using a skeletonized growth series of American alligators (Alligator mississippiensis) of known sex. No statistical support for the hypothesis was found. Furthermore, analysis of a diversity of crocodilian taxa from museum collections revealed similar findings suggesting the alligator results are not taxon specific. Study of well-preserved tyrannosaurid dinosaurs in museum collections showed nearly invariant chevron positioning like that seen in crocodilians. This suggests the usefulness of chevron anatomy for sexing dinosaurs is tenuous. PMID- 16351977 TI - Feeding behavior modulation in the leopard lizard (Gambelia wislizenii): effects of noxious versus innocuous prey. AB - Feeding, a fundamentally rhythmic behavior in many animals, is expected to exhibit modulation with respect to prey type. Using high-speed videography (200 frames(-1)) and kinematic analysis, we investigated prey-processing behavior in the long-nosed leopard lizard (Gambelia wislizenii). The effects of two prey types were examined, innocuous immature crickets (Acheta domesticus) and noxious stinging hymenopterans (honeybees [Apis mellifer] and yellow jackets [Vespula sp.]). Stinging hymenopterans are processed more extensively, with higher gape cycling frequencies, and for a greater duration than are crickets. Generalized tetrapod feeding models were used as a framework to test the hypothesis that gape profile characteristics are modulated in response to prey noxiousness. Cricket processing generally fits the models, but hymenopteran processing departs from typical model parameters. In particular, the SO phase is absent to barely detectable during hymenopteran processing. This likely represents an effect of extrinsic neural input on a centrally directed rhythmic motor pattern, possibly to avoid being stung. Differences in the capture behavior of crickets versus hymenopterans indicate that G. wislizenii assesses prey noxiousness before physical contact with prey and modifies its capture behavior accordingly. These results add to the growing body of evidence that sensory information can play a critical role in shaping stereotyped rhythmic behaviors in non-mammalian tetrapods. PMID- 16351978 TI - The development of the hindbrain afferent projections in the axolotl: evidence for timing as a specific mechanism of afferent fiber sorting. AB - The aim of this study is to reveal the timing and growth pattern of central octavolateral projection development in the Mexican axolotl, Ambystoma mexicanum. In this amphibian species the development of the inner ear occurs first, followed by mechanosensory lateral line organs, and finally by ampullary electroreceptors. Several hypotheses have been proposed about how the development of peripheral organs, including differential projections of the ear, might relate to the development of central projections. Our data suggest that the sequence of maturation of the ear, mechanosensory lateral line, and ampullary electroreceptive organs is closely accompanied by the timed development of the trigeminal, inner ear, mechanosensory lateral line organs, and the ampullary electroreceptor afferent projections in the axolotl. Our data suggest that segregation of central termination within the alar plate is a function of time and space: later forming organs are likely innervated by later forming ganglia that project centrally later and to more dorsal areas of the alar plate that have not yet received any other afferents. Later forming ganglia of the same type may grow along existing pathways of earlier formed neurons. PMID- 16351979 TI - The retro-articular process, streptostyly and the caecilian jaw closing system. AB - Caecilians have two functionally separate sets of jaw closing muscles. The jaw adductor muscles are parallel fibered muscles positioned close to the jaw joint and their lever mechanics suggests they are well suited to rapidly closing the jaws. A second set of muscles, the hypaxial interhyoideus posterior (IHP), levers the jaws closed by pulling on the retro-articular process (RA) of the lower jaw. Models of the lower jaw point out that the angle and length of the RA has a profound effect on the closure force exerted by the IHP. The caecilian skull is streptostylic - the quadrate-squamosal apparatus (QSA) moves relative to the rest of the skull, a condition that seems at odds with a well-ossified cranium. Modeling the contribution of this streptostylic suspension of the lower jaw shows that rotational freedom of the QSA amplifies the force of the IHP by redirecting force applied along the low axis of the lower jaw. Measurements from several species and life stages of preserved caecilians reveal a large variation in predicted bite force (as a multiple of IHP force) with age and phylogeny. PMID- 16351980 TI - Structure, attachment, replacement and growth of teeth in bluefish, Pomatomus saltatrix (Linnaeus, 1776), a teleost with deeply socketed teeth. AB - Tooth replacement poses many questions about development, pattern formation, tooth attachment mechanisms, functional morphology and the evolution of vertebrate dentitions. Although most vertebrate species have polyphyodont dentitions, detailed knowledge of tooth structure and replacement is poor for most groups, particularly actinopterygians. We examined the oral dentition of the bluefish, Pomatomus saltatrix, a pelagic and coastal marine predator, using a sample of 50 individuals. The oral teeth are located on the dentary and premaxillary bones, and we scored each tooth locus in the dentary and premaxillary bones using a four-part functional classification: absent (A), incoming (I), functional (F=fully ankylosed) or eroding (E). The homodont oral teeth of Pomatomus are sharp, deeply socketed and firmly ankylosed to the bone of attachment. Replacement is intraosseus and occurs in alternate tooth loci with long waves of replacement passing from rear to front. The much higher percentage of functional as opposed to eroding teeth suggests that replacement rates are low but that individual teeth are quickly lost once erosion begins. Tooth number increases ontogenetically, ranging from 15-31 dentary teeth and 15-39 premaxillary teeth in the sample studied. Teeth increase in size with every replacement cycle. Remodeling of the attachment bone occurs continuously to accommodate growth. New tooth germs originate from a discontinuous dental lamina and migrate from the lingual (dentary) or labial (premaxillary) epithelium through pores in the bone of attachment into the resorption spaces beneath the existing teeth. Pomatomus shares unique aspects of tooth replacement with barracudas and other scombroids and this supports the interpretation that Pomatomus is more closely related to scombroids than to carangoids. PMID- 16351981 TI - Magic bullets and golden rules: data sampling in molecular phylogenetics. AB - Data collection for molecular phylogenetic studies is based on samples of both genes and taxa. In an ideal world, with no limitations to resources, as many genes could be sampled as deemed necessary to address phylogenetic problems. Given limited resources in the real world, inadequate (in terms of choice of genes or number of genes) sequences or restricted taxon sampling can adversely affect the reliability or information gained in phylogenetics. Recent empirical and simulation-based studies of data sampling in molecular phylogenetics have reached differing conclusions on how to deal with these problems. Some advocated sampling more genes, others more taxa. There is certainly no 'magic bullet' that will fit all phylogenetic problems, and no specific 'golden rules' have been deduced, other than that single genes may not always contain sufficient phylogenetic information. However, several general conclusions and suggestions can be made. One suggestion is that the determination of a multiple, but moderate number (e.g., 6-10) of gene sequences might take precedence over sequencing a larger set of genes and thereby permit the sampling of more taxa for a phylogenetic study. PMID- 16351982 TI - A study of the utility of convergent characters for phylogeny reconstruction: do ecomorphological characters track evolutionary history in Anolis lizards? AB - The reconstruction of phylogeny requires homologous similarities across species. Characters that have been shown to evolve quickly or convergently in some species are often considered to be poor phylogenetic markers. Here I evaluate the phylogenetic utility of a set of morphological characters that are correlated with ecology and have been shown to evolve convergently in Anolis lizards in the Greater Antilles. Results of randomization tests suggest that these "ecomorph" characters are adequate phylogenetic markers, both for Anolis in general and for the Greater Antillean species for which ecomorphological convergence was originally documented. Explanations for this result include the presence of ecomorphologically similar species within evolutionary radiations within islands, some monophyly of ecomorphs across islands, and the existence of several species that defy ecomorphological characterization but share phylogenetic similarity in some ecomorph characters. PMID- 16351983 TI - Phylogeny and evo-devo: characters, homology, and the historical analysis of the evolution of development. AB - The concept of homology continues to attract more and more commentary. In systematic and evolutionary biology the meaning of homology as synapomorphic similarity inherited from a common ancestor has gained wide acceptance over the last three or four decades. In recent years, however, developmental biologists, in particular, have argued for a new approach to, and new definition for, homology that revolves around the desire to make it more process-oriented and more mechanistic. These efforts raise questions about the relationship between developmental and evolutionary biology as well as how the evolution of development is to be studied. It is argued in this paper that this new approach to homology seemingly decouples developmental biology from the study of the evolution of development rather than to facilitate that study. In contrast, applying the notion of historical, phylogenetic homology to developmental data is inherently comparative and therefore evolutionary. PMID- 16351984 TI - A new phase in the new millennium. PMID- 16351985 TI - Message from the president of the Cardiac Society. PMID- 16351986 TI - Outcomes of coronary artery bypass grafting: a 3 year analysis using the Society of Thoracic Surgeons Database. AB - BACKGROUND: Accurate risk factor analysis is a critical element in contemporary cardiac surgical practice. In the USA, the Society of Thoracic Surgeons Database allows institutions and individual surgeons to carry out detailed patient risk assessment and to review their cardiac surgical outcomes in a comparative fashion. METHODS: To evaluate outcomes of isolated coronary artery bypass grafting, data from all patients operated upon at the Alfred Hospital, Melbourne, Australia, over a 3 year period were entered into the Society of Thoracic Surgeons Database. RESULTS: Our results (mortality and morbidity) compared favourably with those contained within this large international database. CONCLUSION: It is hoped that a similar Australasian database can be established to facilitate a meaningful local risk assessment and a comparative analysis of outcomes of cardiac surgical procedures. PMID- 16351987 TI - Surgical aspects of endocarditis. AB - Endocarditis represents a difficult medical problem that must occasionally be treated surgically. Constitutional symptoms of infection are important and are usually the reason that the patient seeks medical attention. Fever is the most common sign of infective endocarditis and a heart murmur, which changes in character or is new, is a significant hallmark. The diagnosis for infective endocarditis is made by high index of suspicion in a patient with valvular heart disease or a prosthetic heart valve and in the presence of fever and a cardiac murmur. A positive blood culture is the hallmark of the diagnosis. The absolute indications for operative intervention are congestive heart failure, unstable prosthetic valve, uncontrolled infection, and relapse after optimal therapy (prosthetic valve). Relative indications for operative intervention are perivalvular extension of the infection, staphylococcal infection of a prosthesis, persistent fever (culture negative), large vegetation, or relapse after optimal therapy (native valve). The principles of surgical management are to remove all infected tissue by thorough debridement back to normal tissue. This is combined with replacement of damaged valves and repair of associated defects. The mortality after operation for infective endocarditis is 15-20%. Late survival after operation for infective endocarditis on a native heart valve is 70-80% at 5 years. Survival falls to 50-80% at 5 years for surgery on an infected prosthetic heart valve. PMID- 16351988 TI - Cardiac surgery and neurological injury in children. AB - The freedom of children from vascular disease makes them an ideal population in which to study the deleterious effects of cardiopulmonary bypass. Cerebral embolic injury arises intraoperatively from air in the cardiac chambers and monitoring lines, particularly those in the left atrium. Postoperative emboli are more common in patients with single ventricle physiology, especially following a bidirectional Glenn shunt or Fontan procedure. Poor cannula selection or placement during bypass can cause maldistribution of the perfusate and cerebral injury. Postoperative hyperthermia can cause brain damage. Results of prospective and retrospective studies of neurological and developmental outcome after paediatric cardiac surgery at the Children's Hospital, Boston, have led to the following inferences: the use of circulatory arrest should be minimised; the pH stat strategy is preferred for cardiopulmonary bypass over the alpha stat strategy. A higher haematocrit on bypass, for example 30%, may be preferable to 20%; postoperative fever should be strenuously avoided. Surgeons should be aware that cardiopulmonary bypass parameters can have subtle but important effects on late developmental outcome. PMID- 16351989 TI - Prospective development of a cardiac surgical registry. AB - Interest has recently been expressed in developing an Australian adult cardiac surgical registry. Complete national registries of adult cardiac surgery have already been established in many European countries, the USA, Canada and elsewhere. Participating centres contributing to a national registry benefit by being able to benchmark themselves against norms for their particular country. A risk-adjusted database can help surgeons advise their patients of the chances of a good operative outcome. For a surgeon or a surgical unit, the only way to obtain a relevant risk model is to use their own data, and data from units in their particular country. It is also useful to have comparative data from other national registries to compare one's own country with international benchmarks. Since 1996, the European Cardiac Surgical Registry (ECSUR) has put considerable effort into producing unified datasets, harmonised with each other for worldwide use. In 1997, ECSUR launched a minimum cardiac surgical dataset. The worldwide launch of the full international adult cardiac surgical dataset is scheduled for July/August 2000. This dataset would be highly useful for application in Australia. The ECSUR organisation has the capability to analyse data from other countries and could perform this for Australia if requested. However, a better approach would be a national centre in Australia. Funding for national registries around the world has been obtained from Ministries of Health, participating surgical centres, and surgical software vendors. If an Australian national registry is indeed established it will find a ready-made, highly appropriate international cardiac surgical dataset sponsored by ECSUR and the Society of Thoracic Surgeons waiting for adoption by Australia. PMID- 16351990 TI - Thrombus on an ASD closure device: a call for caution. AB - Use of per-catheter atrial septal defect closure devices is becoming increasingly widespread. We report a case of left atrial thrombus formation on a StarFLEX device raising concerns regarding the general use of these devices and as a means of preventing paracloxical embolism in particular. PMID- 16351991 TI - Successful two patch repair of the aortico-left ventricular tunnel. AB - A 7-month-old boy with aortico-left ventricular tunnel arising from the right coronary sinus underwent surgical correction using the two patch technique. The diagnosis was confirmed by 2D and Doppler echocardiogram alone. Patch closure of both the aortic and ventricular ends of the tunnel is recommended in order to prevent postoperative aortic regurgitation and recurrence of the lesion. PMID- 16351992 TI - Acute laryngeal oedema and the yellow nail syndrome. AB - A complication of yellow nail syndrome after pleurodesis is presented. After endotracheal extubation the patient developed delayed acute laryngeal oedema requiring re-intubation. We postulate that the combination of lymphatic defect and airway trauma from the double lumen endotracheal tube were contributory to the oedema. If endotracheal intubation is performed, these patients should be observed for 48 h. PMID- 16351993 TI - Dendritic cells in aortocoronary saphenous vein bypass grafts. AB - Dendritic cells are specialised leucocytes responsible for capturing and presenting antigens to T lymphocytes, which in turn mediate immune responses in various pathological conditions. The observations in this study demonstrate that dendritic cells are present in stenotic aortocoronary saphenous vein bypass grafts, which suggest that these cells may be involved in inflammatory actions which may subsequently contribute to graft failure. PMID- 16351994 TI - Coronary artery bypass surgery. PMID- 16351996 TI - Burgeoning atherosclerosis in Asia. PMID- 16351995 TI - A mighty people who never wrote a word. PMID- 16351997 TI - The stentless xenograft aortic valve: the wheel turns around. AB - BACKGROUND: A brief overview of the historical pathways of both stented and stentless porcine xenografts is presented in order to understand the return to and continuing clinical use of stentless devices. In addition, 7-11 years of durability with various models of stentless porcine valves has now accumulated and is beginning to be of relevance in determining the future place of this xenograft. Stentlessness and anticalcium agents, coupled with the poor results of stented xenografts in certain patient groups, have led to a resurgence of the clinical use of stentless xenograft valves for aortic valve replacement. An overview of the present state and future of stentless valves is given. METHODS: At both The Prince Charles Hospital and St Andrew's War Memorial Hospital, Queensland, Australia, 307 patients have received the Model 300 CryoLife-O'Brien stentless composite aortic xenograft from December 1992 to February 2000. Associated procedures were required in 56% of patients (mostly coronary artery bypass, mean 2.4 grafts, in 144 patients (47%) and left ventricular myomectomy in 34 patients (11%)). RESULTS: The hospital mortality (four early deaths) has been 1.3 +/- 1% (CL 95%) and the follow-up 100% for this analysis. The mean patient age was 73 years (range 57-89 years with 16% being 80 years and over). Morbid events have included six perivalvar leaks: four trivial and identified only on echo Doppler (no clinical murmurs) and two patients requiring reoperation at 10 days and 12 weeks with simple successful repair verified on subsequent echocardiograms. Of the 307 patients over the 7 year period, three valves only have been explanted, two for endocarditis at 1.5 and 3.5 years and one for possible technically induced structural failure at 15 months (probable needle damage). With this exception, there has been as yet no other intrinsic leaflet failure. Four early thromboembolic events (4 days-5 weeks) in patients with atrial fibrillation (no anticoagulants used postoperatively with the first 80 patients) constituted the important early morbid events. Late mortality of this elderly patient cohort has occurred in 27 patients over 7 years of maximum follow up. One death (endocarditis) has been valve related at 5 years. Serial echocardiography (some 700 echoes in the study of this valve) has demonstrated a mean gradient of 7-9 mmHg with a very low incidence of trivial incompetence (96%) on Doppler examination with implant valve sizes ranging from 21 to 29 mm. One patient had significant regurgitation requiring reoperation. There has been no progression of either incompetence or stenosis of the remaining patients in this follow-up, now into the eighth postoperative year. CONCLUSION: The early and intermediate results appear excellent in this elderly patient cohort. Nevertheless, important surveillance is obviously required to determine the durability at 10-12 years, a crucial time when stented porcine xenografts began to show an obvious failure rate from structural deterioration, in the middle and elderly aged patient cohort. An attempt is made to outline the future of this type of stentless xenograft and to justify that its cautious use should probably be extended down to the over 50 year age patient cohort. PMID- 16351998 TI - Prophylactic intravenous mannitol and normal saline in patients with poor renal function prior to cardiac surgery: time for a multicentre trial? AB - BACKGROUND: Postoperative acute renal failure requiring dialysis has a poor prognosis, which has remained unaltered for 50 years. Therefore, in cardiac surgical patients at increased risk of postoperative oliguric acute renal failure (preoperative serum creatinine >0.13 mmol/L), we assessed the use of prophylactic intravenous (i.v) 20% mannitol and normal saline therapy in addition to traditional methods of therapeutic renal support. METHODS: Seventy-five patients with a mean preoperative serum creatinine of 0.192 mmol/L received i.v. 20% mannitol and normal saline pre-, intra- and postoperatively. This treatment was continued postoperatively until serum creatinine returned to baseline. RESULTS: No patient required dialysis, no patient died, developed a myocardial infarction nor a stroke in the first 30 days post-surgery. Serum creatinine and urea increased to a mean peak on day 3 of 0.233 +/- 94 and 24.6 +/- 13 mmol/L, respectively. However, all patients maintained a high urine output (>2 L/day) and no patient required haemodialysis. CONCLUSION: Therapy with i.v. 20% mannitol and normal saline appears safe and effective in maintaining a diuresis and may avoid the need for dialysis. A randomised controlled trial of this treatment in patients at increased risk of postoperative acute renal failure is warranted. PMID- 16351999 TI - A brain dead donor model of porcine orthotopic cardiac transplantation for assessment of cardiac allograft preservation. AB - BACKGROUND: We aimed to develop a large animal model of orthotopic cardiac transplantation, incorporating donor brain death, to assess new methods of preservation of the donor heart. METHODS: Brain death was achieved in the donor pig by inflation of a 20 cc subdural balloon 1 h prior to harvest. The donor heart was stored for 6 h with conventional hypothermic ischaemic preservation. It was then transplanted orthotopically into the recipient pig using the Lower and Shumway technique. One hour after reperfusion, the transplanted heart was weaned from cardiopulmonary bypass with dobutamine support. Dobutamine support was continued for up to 4 h, if required. After 6 h of physiological and biochemical evaluation, the recipient was euthanased and the heart excised for histological assessment. RESULTS: All pigs experienced the classical haemodynamic changes associated with brain death. This resulted in the release of Troponin I, consistent with myocardial injury. The donor operation was successfully completed in 11 out of 13 pigs. Six out of 11 transplanted hearts were successfully weaned from cardiopulmonary bypass, but required ongoing dobutamine support. CONCLUSIONS: This porcine model of orthotopic cardiac transplantation is a relevant and practical large animal model for the assessment of new methods of preservation of the donor heart. PMID- 16352000 TI - How to write a paper for publication. AB - Engaging in the scientific publication process can be for both altruistic and egotistical reasons; publication advances the state of scientific knowledge while advancing your institution and your career. Writing for publication means setting aside a location and time dedicated entirely to the process of planning and writing. It is easiest to begin with the Methods section, then the Results, followed by the Discussion, which is the most challenging part of a paper. A realistic assessment of the value of the article will determine the level of journal into which it is likely to gain acceptance. If your article is rejected by a journal, be consoled by the fact that 50% of articles that are initially rejected are eventually published. Following the steps outlined here can reduce the daunting task of writing to one of manageable proportions and can help overcome the mental block and procrastination that all of us have experienced when we set out to write a scientific paper. PMID- 16352002 TI - The Olympian challenge: science, sport and society. PMID- 16352001 TI - Aortic dissection associated with Campylobacter aortitis. AB - Infective aortitis and associated mycotic aneurysms are uncommon clinical entities and most reported cases are associated with Salmonella species. We report successful surgical and medical treatment of a mycotic aneurysm of the aorta which presented as acute pericarditis and may have been caused by Camplyobacter jejuni. PMID- 16352003 TI - Three-dimensional echocardiography: research toy or clinical tool? AB - Conventional 2D echocardiography is an excellent qualitative imaging method, but its use for quantitation is limited by test-retest reproducibility of image planes. The increasing sophistication of medical treatments for left ventricular dysfunction, hypertension and valvular heart disease has created the need for accurate and reproducible measurements of chamber dimensions. Similarly, improvements in valve repair and catheter-based interventions for valve lesions and septal defects have created the need for better visualisation of cardiac structures. The use of 31) echocardiography may decrease variability both in the quality and interpretation of complex pathology among investigators. Three dimensional echocardiography is achieved by using a 3D spatial registration device with a conventional 21) scanner, or by using a high-speed, phased-array real-time scanner. The latter are still developmental, so that the technique currently requires use of a 21) scanner, combined with a 31) spatial coordinate system, which may be external or internal to the scanning transducer. An external system permits data acquired from several cardiac windows to be integrated and reconstructed. Image reconstruction is performed using a wire-frame model or surface rendering. Wire-frame models are formed by manual or automatic connection of boundary data points; this approach uses fewer data points than rendering, can be rapidly processed and is sufficient for quantitative analysis. Surface rendering uses lighting and shading applied to a wire-frame model to produce a realistic 31) display, which may be useful for surgical planning and increasing understanding of anatomic relations. Three-dimensional echocardiography yields more accurate measurements of ventricular volume and function, as well as new measurements such as infarct area. With increased reproducibility and reliability, 3D echocardiography may well prove to be the essential tool required for the serial follow up of left ventricular mass and volume. PMID- 16352004 TI - Arterial revascularisation with two or more arterial conduits. AB - In the last decade arterial revascularisation has become more popular due to the routine use of the left internal mammary artery (LIMA). Left internal mammary artery grafting to the left anterior descending (LAD) coronary artery is widely recognised as the most important single determinant of improved results from coronary revascularisation. However, a question remains as to whether results can be further improved by using other arterial grafts in territories other than the LAD, or increasing the number of arterial anastomoses per patient. Although long term results do not conclusively prove that arterial anastomoses to vessels other than the LAD increase the quality of late results, there are no results to disprove this hypothesis. In our institution we ensure that all mammary grafts are placed on the left coronary system. To achieve this we use skeletonised mammary conduits to increase graft length, and Y grafts to minimise the number of aortic anastomoses. We use the gastroepiploic artery as an in situ graft. With the radial artery we avoid aortic anastomoses by joining the radial artery to the internal mammary artery. With the increased use of skeletonised internal mammary arteries we use the inferior epigastric artery less frequently. This strategy for arterial revascularisation is based on results of 2236 patients undergoing myocardial revascularisation between October 1991 and June 2000. Of these, 75% had total arterial revascularisation where the early mortality was 2% and survival 93% at 7 years. We conclude that total arterial revascularisation using bilateral mammary grafts supplemented by other arterial conduits provides optimal results for myocardial revascularisation. PMID- 16352005 TI - The heart as a target for oestrogens. AB - Observational studies have consistently shown a markedly decreased risk of cardiovascular disease in postmenopausal women when treated with oestrogens. This review discusses plausible mechanisms for the physiological effects of oestrogens in healthy and diseased hearts. Oestrogens have well-documented effects on blood lipids and the regulators of the cardiovascular system, which should reduce risk. In addition, the heart is a primary target for oestrogens with functional oestrogen receptors in the coronary vasculature and on cardiac myocytes and fibroblasts. Rapid oestrogen effects include vasodilatation and anti-arrhythmic effects by actions on ion channels, and some of these effects may be pharmacological rather than physiological. Longer term responses to physiological levels of oestrogen include an increased expression of nitric oxide synthase in myocytes and endothelial cells as well as proinflammatory and pro-arrhythmic effects. Oestrogens induce growth of non-proliferating fibroblasts but inhibit the replication of proliferating fibroblasts. In contrast to the observational studies, two randomised, controlled studies of oestrogen and progestins in postmenopausal women with coronary heart disease have now shown increased coronary events, especially in the first year of study, and no change in the progression of coronary atherosclerosis. Further studies of the complex effects of oestrogens on healthy and diseased animal models are essential. Large clinical trials of the newer selective oestrogen receptor modulators to lower cardiovascular risk in both males and females should be considered as a priority. PMID- 16352006 TI - Salmonella prosthetic valve endocarditis. AB - We report the case of a 69-year-old man with an aortic valve bioprosthesis in whom a diagnosis of Salmonella typhimurium prosthetic valve endocarditis was made. During the prolonged hospital admission early aggressive antibiotic therapy and early valve replacement surgery were the deciding factors enabling him to survive this otherwise lethal condition. PMID- 16352007 TI - Transfusion-associated graft versus host disease postcardiac surgery. AB - We describe a 75-year-old male who suffered transfusion-associated graft versus host disease postcoronary artery bypass graft surgery in Australia. We discuss pathogenesis, incidence, mortality, treatment options and preventative suggestions. PMID- 16352008 TI - Peer review and quality assurance in cardiac surgery in Australia. AB - There is increasing community and government interest in determining the quality of medical performance in Australia. This means that any lead taken by the Australasian Society of Cardiac and Thoracic Surgeons (ASCTS) in defining and adopting performance indicators for the profession is appropriate and timely. The ASCTS Peer Review Committee has examined the issue of quality management, and presents here several key indicators and a draft program of remediation which, if adopted, will facilitate the achievement of the highest and most uniform surgical standards for Australia. The Committee recommends the monitoring of just one procedure initially: coronary artery surgery. The Committee sees the quality management program as involving both surgeons and hospital-based quality assurance teams. Ultimately, the profession will gain from performance measurement by being able to minimise inappropriate variations in practice. The community will benefit by having national and international best practice in cardiothoracic surgery, with surgeons being able to identify more easily those procedural changes that result in superior as well as inferior outcomes. PMID- 16352009 TI - Legal issues relating to a cardiac surgery database. AB - The proposed Cardiac Surgery Database project is a venture shared between the Victorian Department of Human Services and the Australasian Society of Cardiac and Thoracic Surgeons (ASCTS). The aim is to provide a readily available and comprehensive information base on cardiothoracic procedures carried out in Victoria, with a view to raising the standards of cardiothoracic surgery, and to facilitate long-term patient care. Important issues for consideration are ownership and protection of the Database and the information within it, as well as the patient consent process, privacy and patient access to data, commercial use of the Database for ongoing funding, risk management, and protection of the ASCTS and surgeons from litigation. PMID- 16352010 TI - The potential role of the Australian Institute of Health and Welfare in a National Cardiac Surgery Database. AB - The Australian Institute of Health and Welfare has responsibility for developing national health information and collections and is thus a candidate for building and operating a National Cardiac Surgery Database. Many functional characteristics place the Institute in a favourable position for establishing and maintaining a national patient-based database of cardiac surgery. These include its experience in the field of database management, its record of objectivity and its capacity to provide data protection. In addition it has significant experience in monitoring cardiovascular diseases, their risk factors, treatments and outcomes through its National Centre for Monitoring Cardiovascular Disease. The Institute, a statutory body operating within the Federal Government's health portfolio, is independent of policy, administrative and regulatory functions within that portfolio. The health-related databases managed by the Institute include those that record national statistics on insulin-treated diabetes, hospital separations, cancer and death. The Institute also maintains, on behalf of the National Heart Foundation, a cardiac surgery register based on aggregated data from cardiac units. This register is not patient based as the new database would be. With this range of experience, the Institute sees itself well placed to contribute to the development of this important project. PMID- 16352011 TI - A National Cardiac Surgery Database: current achievements and future challenges. PMID- 16352012 TI - Why Australia needs a Cardiac Procedures Database. AB - The development of a national cardiac procedures database is in the interest of many groups: cardiac surgeons and interventional cardiologists, participating hospital units, patients, governments, health insurance funds and industry. To date, information about short-term outcomes of coronary interventions has been fragmented and uncoordinated at state and national levels. Long-term outcomes have not yet been systematically addressed in Australia. A highly desirable objective is the development of a dataset of information that is pertinent to the Australian context, with common minimum datasets for surgery and percutaneous intervention. Information will gain added value if it is linked to other national databases such as the National Death Index. Also, risk adjustment should be used to improve the value of outcome data. Development of a 'common' Cardiac Procedures Database, in turn, should lead to improved health policy and better health management in an area that currently runs at significant cost. Thus, provided due care is taken to protect the privacy of stakeholders, maintain quality control and entry of only valid information into the database, the resulting outcome will benefit all interested parties; clinicians, hospitals, patients and the wider Australian community. PMID- 16352013 TI - Towards a Statewide Cardiac Surgery Database: the Victorian initiative. AB - The establishment of the Australasian Society of Cardiac and Thoracic Surgeons (ASCTS) at the beginning of the 1990s gave rise to an early proposal for the development of a cardiac surgical database. The Victorian Government gave its in principle backing to a statewide database project from late 1997. Release of a promised dollar 200,000 grant for development was contingent on the ASCTS being able to fulfil several initial requirements. These were: (i) the provision of an indicative annual report outlining performance against a series of clinical indicators; (ii) an indicative peer-review process; and (iii) the addressing of issues surrounding intellectual property and ownership of any database that was developed. In outlining a strategy to be used in developing and implementing the project plan, it was determined by the project development team that all Victorian cardiac surgical units were, in fact, collecting similar data, but were using different software and analytical tools. The Cardiovascular Disease Prevention Unit at the Baker Medical Research Institute in Melbourne has developed a model of mortality and risk-adjusted complications out of the pooled Victorian data. This, it is hoped, will be a forerunner of a major achievement in the monitoring of cardiac surgery procedures nationally, and the attainment of national and international best practice. PMID- 16352014 TI - Developing performance indicators for cardiac surgery: a demonstration project in Victoria. AB - Six Victorian cardiac surgical units pooled data in order to undertake a demonstration project aimed at developing performance indicators to assess outcomes following cardiac surgery. The outcome of the project was an indicative report for the purpose of monitoring surgical performance indicators in a format suitable for: (i) the general public; (ii) the Victorian State Government; and (iii) the participating units and surgeons. Each participating cardiac surgical unit had an existing database used for recording information from each procedure. A request was made to each unit to extract a subset of data from all cases entered over the past 5 years. The proposed list of performance indicators included surgical mortality (within the period of admission for surgery), complication rates (including sternal infection, postoperative myocardial infarction, postoperative stroke, haemorrhage requiring return to theatre), and length of hospital stay. A model was developed from the data and used to provide risk-adjusted measures of hospital performance. Cases from five cardiac surgical units (n = 10 715) were included in the final analysis. A risk-adjusted model (including age, sex, diabetes, hypertension, smoking, procedure type, urgency of procedure) was developed for surgical mortality. Performance indicators for coronary artery bypass graft surgery, including mortality, sternal infection rate and length of hospital stay are presented. From the available data, performance indicators for cardiac surgery in Victorian hospitals compared favourably with international benchmarks. This project has demonstrated that prospective data collection using a standardised system could readily produce local risk adjustment models for cardiac surgery to aid in developing appropriate performance indicators. PMID- 16352015 TI - Implementation of a Victorian Cardiac Surgery Database. AB - The Victorian Cardiac Surgery Database project will be jointly developed and delivered by Melbourne Information Management and the Baker Medical Research Institute. After a pilot phase involving one hospital, all Victorian public hospitals with cardiac surgery units will provide data for developing and testing the Cardiac Surgery Database. In the initial proposal, the period of data collection is from 1 April 2001 to 31 March 2002. Issues being addressed in preparation for the implementation phase are those of ownership, management and administration of the Database as well as taxation, liability and governance. Completeness and accuracy of the data collection process and the maintenance of confidentiality are prime considerations in the implementation phase of the project. PMID- 16352016 TI - A National Cardiac Surgery Database: why, how and when? AB - As users of large amounts of public funding, surgeons are increasingly being required to justify their activities. The provision of accurate risk-stratified data on cardiac surgical procedures and their outcomes allows for government review while also providing a means of achieving potential improvements to strategies for the management of higher risk patients. In addition, accurate data will allow for the appropriate assessment of results that fall outside acceptable benchmark standards. A management strategy may then be implemented, following peer-review processes, to examine the outlying results on an anonymous basis. The dataset currently employed in the Victorian Database is derived from USA and UK models, and it will be used for the development of a national database. This project has Federal Government support in principle, and, it is hoped, its eventual financial backing. PMID- 16352017 TI - Data analysis for the National Cardiac Surgery Database. AB - The planned Australian National Cardiac Surgery Database is likely to have a number of positive outcomes, including increased patient satisfaction, improved quality assurance and increased economic efficiency. In relation to cardiac surgery, performance indicators associated with the commonly performed procedure of coronary artery bypass surgery will be used for peer review and to measure outcomes. Several different risk-adjusted models are available for analysing national databases. However, the potential weaknesses of database analysis are lack of both compliance and data validity. A number of other major issues, such as location of the data analysis centre, who will hold authority over data accuracy, and the security of and access to the Database, must also be considered when setting up the National Database. Overall, however, the benefits of a national database will be enormous. Cardiologists and cardiac surgeons will benefit from a disease-based registry with shared common definitions. In addition, the provision of such a database will represent a crucial step towards developing national strategies for treating heart disease. PMID- 16352018 TI - National cardiac databases for Australia. PMID- 16352019 TI - Univentricular repair in children under 2 years of age: early and midterm results. AB - BACKGROUND: Encouraging results have been obtained from early univentricular repair. Our study was designed to assess the impact of young age (less than 2 years) on the outcome of univentricular repair. PATIENTS AND METHODS: Between January 1992 and December 1998, 65 out of 311 patients undergoing univentricular repair at the All India Institute of Medical Sciences, New Delhi, were less than 2 years of age. We compared these 65 carefully selected patients (group 1) with the 246 patients who were more than 2 years of age (group 2). Since 1994, all patients of both groups had a fenestration of the intra-atrial baffle. RESULTS: The early mortality rate was 9.2% in group 1 and 7.7% in group 2. The overall Fontan failure rate was 12.3% in group 1 and 12.2% in group 2. In group 1, a higher incidence of Fontan failure was noted in patients with suboptimal weight, non-tricuspid atresia morphology, nonfenestrated Fontan and those who did not comply with more than two of Choussat's criteria, but the values were not statistically significant. Aortic cross-clamp time of more than 60 min was the only and highly significant predictor of Fontan failure (P < 0.01). The overall effusion rate was 24.05% in group 1 and 27.98% in group 2. In group 1, patients weighing less than 10 kg (P = 0.0007), without fenestration of the atrial baffle (P < 0.05) and with systemic ventricular dysfunction (P < 0.001), systemic ventricular end-diastolic pressure of more than 12 mmHg (P < 0.001), mean pulmonary artery pressure of more than 15 mmHg (P < 0.001) and aortic crossclamp time of more than 60 min (P < 0.01), were all found to be significant risk factors of pleural effusion. Pulmonary artery distortion needing reconstruction did not increase the Fontan failure or effusion rates. Oxygen saturation ranged from 85 to 94% (mean 89%) in patients having a functioning fenestration at a mean follow-up period of 30 months. The actuarial survival at 84 months was 90 +/- 0.04% in group 1 and 88 +/- 0.02% in group 2. CONCLUSIONS: Our results suggest that carefully selected patients under 2 years of age are suitable candidates for one-stage univentricular repair and that survival is not significantly different from that of older patients. Routine fenestration of the intra-atrial baffle is an option available to ameliorate morbidity and obviate mortality. Non-compliance with more than two of Choussat's criteria appears to be additive in unsatisfactory outcome. PMID- 16352020 TI - Angina pectoris and coronary artery disease in severe isolated valvular aortic stenosis. AB - BACKGROUND: Angina pectoris has long been recognised as one of the principal symptoms of severe aortic valve stenosis (AS), even in patients without significant coronary artery disease (CAD). However, controversy exists concerning the prevalence of angina pectoris and associated CAD in such patients. OBJECTIVE: To determine the prevalence of CAD detectable by angiography and its relation to angina pectoris and coronary risk factors in patients with severe AS. PATIENTS AND METHODS: All patients with symptomatic AS who had undergone aortic valve replacement and preoperative cardiac catheterisation at the Austin and Repatriation Medical Centre between 1 January 1986 and 31 May 1996 were retrospectively analysed. Those patients with multiple valve disease, aortic regurgitation of grade 2 or more in severity, or who had had prior coronary artery or valve surgery were excluded from this analysis. RESULTS: A total of 328 consecutive patients with severe AS (242 men and 86 women; mean age 72 years, range 39-84 years) were studied. Significant CAD (reduction in luminal diameter > or = 50%) was found in 162 patients (49.4%). Typical angina was present in 74.7% of these 162 patients but it was also found in 44.6% of the 166 patients without obstructive CAD. Of the patients without angina (n = 133), 30.8% had significant CAD. By multivariate logistic regression, we have identified seven significant predictors for CAD among severe AS patients. Five factors increased risk. Expressed as odds ratio with 95% confidence interval, these included: (i) age in years (1.07; 1.04-1.11, P = 0.001); (ii) male gender (2.09; 1.14-3.80, P = 0.016); (iii) angina pectoris (3.19; 1.89-5.37, P < 0.001); (iv) history of myocardial infarction (2.87; 1.38-5.97, P = 0.005); and (v) peripheral vascular disease (2.28; 1.28-4.05, P = 0.005). Factors associated with decreased likelihood of CAD were serum high density lipoprotein (HDL) cholesterol (0.58; 0.34-0.71, P = 0.002) and peak systolic gradient across the aortic valve (0.97; 0.95-0.99, P = 0.0113). CONCLUSION: Coronary arteriography can probably be omitted for a patient with severe AS if that patient has no symptoms of angina and has no risk factors known to increase its incidence. PMID- 16352021 TI - Asia-Pacific collaboration on coronary heart disease risk factor intervention: study design and methods. AB - BACKGROUND: Age-adjusted death rates for coronary heart disease (CHD) have been decreasing in populations of developed countries. At the same time, CHD in the Asia-Pacific region appears to be increasing in parallel with the 'Westernisation' of diet and lifestyle. More epidemiological information is needed from the Asia-Pacific region in order to be able to predict trends in CHD and to plan resources for treatment. In this paper we outline the study design of a survey of coronary heart disease risk factors in Asia-Pacific countries. AIMS: To ascertain and compare in 10 countries: (i) rates of measurement of risk factors; (ii) levels of risk factors, and (iii) management of risk factors. METHODS: A retrospective survey was conducted in 180 randomly selected hospitals using a population-weighted sampling frame. Participating countries were Australia, Indonesia, Japan, Korea, Malaysia, New Zealand, the Philippines, Singapore, Taiwan and Thailand. Data covered a 6-month period from the patient's admission to hospital with myocardial infarction or unstable angina. Data included measures of hypertension, smoking, obesity, diabetes, dyslipidaemia and family history of coronary disease. The study design incorporated rigorous quality-assurance methods to ensure reliability. CONCLUSIONS: The study will provide data comparable to those from studies in Europe and the USA. Together, these studies will provide an international perspective on coronary risk factors. PMID- 16352022 TI - Cardiac angiosarcoma: too little known, too late treatment or just too bad a tumour? AB - Cardiac angiosarcoma is a rare tumour. Current imaging techniques (magnetic resonance imaging, MRI; computed axial tomography, CAT; 2-D echocardiography), although useful in delineating the extent of tumour involvement, do not correlate well with intraoperative findings of resectability. We report a case were palliative surgical resection was technically possible, contrary to expectations from CAT and MRI findings. However, the patient was clinically in extremis, with advanced ventricular dysfunction, and died. Despite the short-term risk involved in surgery, if palliative resection is possible, a multidisciplinary approach with adjuvant chemotherapy and radiation can result in mid-term survival. PMID- 16352023 TI - Myocardial infarction due to saphenous vein graft compression by an extracardiac mass. AB - We present an unusual case of a patient with an inferior infarct caused by saphenous vein graft compression by an extracardiac malignant spindle cell tumour, a mechanism of myocardial ischaemia rarely described. This case also illustrates how chest X-ray and angiography can be useful for making an early diagnosis of a cardiac mass. PMID- 16352024 TI - Walt Lillehei written by George Stirling in Heart, Lung and Circulation (2000; 9: 27-9). PMID- 16352025 TI - Heart valve surgery in China: yesterday and today. AB - The past 20 years have seen rapid development in heart valve surgery in China. By the late 1990s, there were 6000 heart valve operations performed each year. Statistical analysis has shown that rheumatic heart disease is still the leading cause of valvular damage leading to surgery, as it had been 40 years before. The progressive fibrosis, sclerosis and calcification of the mitral valve that characterises rheumatic heart disease caused high mortality for all forms of mitral valve surgery in China in the 1960s. At that time, the introduction of closed mitral commissurotomy, initially highly effective in alleviating symptoms, was later found to result in re-stenosis in a significant cohort of patients. This was progressively replaced with open mitral commissurotomy. Today, mitral valve replacement represents 60-70% of valvular replacement procedures, followed by double-valve (mitral and aortic) replacement (20-25%). It has been shown both in China and elsewhere that careful selection of patients for an absence of mitral calcification leads to higher success rates for surgery. Heart valve replacement surgery in China now attains international standards in terms of the numbers of cases and surgical outcomes. Further long-term data collection and analysis are essential to aid the further development of the field. PMID- 16352026 TI - Status of paediatric cardiac surgery in China. AB - Epidemiological data show that in China, approximately 100,000 neonates annually are born with congenital heart disease, an incidence equivalent to that found internationally. However, surgical treatment is available to only one-fifth of these individuals. Access to surgery is generally becoming more readily available in the urbanised coastal areas than in the rural interior. Over 1000 procedures annually are performed at the two largest facilities, the Fu Wei Hospital, Beijing, and the Shangai Children's Medical Centre. The 300 paediatric cardiac surgery facilities across China perform approximately 30,000 operations each year, with the quality of treatment varying according to location and procedure type: data on Tetralogy of Fallot correction, for example, show that standards are equivalent to those seen internationally, with a mortality rate of 1.1-5%. For other procedures, although outcomes have improved since the beginnings of Chinese paediatric cardiac surgery in the 1970s, there is still a process of quality improvement, especially for the more severe and complex disorders. PMID- 16352027 TI - Trends in beating heart coronary artery bypass grafting with the octopus: experience from 300 cases. PMID- 16352028 TI - Result of heart transplantation waiting list under extracorporeal membrane oxygenation support: experience in National Taiwan University Hospital. PMID- 16352029 TI - Current status of minimally invasive coronary artery bypass grafting in China. AB - A rapid increase in conventional coronary artery bypass grafting (CABG) has been seen in China over the past 10 years. Over the past 4 years, increasing numbers of Chinese surgeons have been exploring new techniques to simplify the operation for the surgeon and the patient. Between 1996 and 2000,117 patients had CABG off pump (OPCAB) at Fu Wai Hospital, Beijing. OPCAB procedures have also been reported from several other centres around China. Minimally invasive CABG has been performed in at least four centres in China. Video-assisted CABG and combined CABG angioplasty techniques are also being explored at Fu Wai Hospital. PMID- 16352030 TI - Historical evolution of minimally invasive coronary surgery. PMID- 16352031 TI - Thoracic aortic aneurysm and dissection: surgical results and experience of 428 cases. PMID- 16352032 TI - Video-assisted lobectomy: Indications and technique. PMID- 16352034 TI - Review of coronary artery surgery in China. AB - The 30-year history of coronary artery surgery in China has shown rapid development, particularly in the past 10 years. By 1994, approximately 100 coronary bypass grafting procedures were performed nationally, with mortality rates halving between 1992 and 1993. By 1998, more than 1000 operations had been performed, with mortality rates falling to less than 1% at Fu Wai Hospital, Beijing, but with varying results across the country. Figures for 1995 from Fu Wai Hospital show that the use of the left internal mammary artery was favoured in 65% of patients, with the radial artery used in less than 10%. Changes to the presenting patient profiles have been noted across time, with the incidence of hypertension and diabetes increasing and with severe ventricular dysfunction decreasing. Off-pump and minimally invasive coronary artery bypass grafting were introduced to China in 1996 with encouraging early results. PMID- 16352035 TI - Present status and future of cardiopulmonary bypass in China. AB - Twenty Chinese hospitals specialising in cardiopulmonary bypass (CPB) carrying out 13 274 procedures were selected for analysis. The technique of CPB was used during surgery for cyanotic and non-cyanotic congenital heart disease, valve replacement, coronary bypass grafting, operations on the great vessels and emergency procedures. A wide variety of CPB techniques, equipment and monitoring procedures were in use across the country. There is a need for standardisation of equipment and procedures, for broadening the skill base of perfusionists, and for the introduction of professional standards to the field whose personnel come from a variety of backgrounds. PMID- 16352036 TI - Angiotensin II: a proven therapeutic target with some mechanistic mysteries. PMID- 16352037 TI - Low serum S100beta protein levels following coronary artery surgery with or without cardiopulmonary bypass. AB - BACKGROUND: S100beta protein has been used as a serum marker of brain injury during cardiac surgery. Previous studies may have been confounded by the re transfusion of shed mediastinal blood, which has a high concentration of S100beta. AIM: To examine serum S100beta levels in coronary artery surgery patients in whom re-transfusion of shed mediastinal blood was avoided, and to compare levels with and without cardiopulmonary bypass (CPB). METHODS: Serum S100beta levels were measured preoperatively, pre-heparin, post-protamine and 24 h postoperatively in 31 patients undergoing elective coronary artery surgery with (n = 14) or without (n = 17) CPB. The postoperative values were compared to their preoperative controls using two-tailed paired t-tests. RESULTS: There was a minor increase in serum S100beta post-protamine in the CPB group only (0.41 ng/mL; P < 0.01). All other levels in both groups were within normal limits. CONCLUSIONS: Coronary artery surgery without CPB is not associated with an increase in serum S100beta. The observed increase in the CPB group was four- to ninefold lower than levels previously reported. A possible explanation for the lower level was the avoidance of retransfusing shed mediastinal blood. If elevated S100beta levels are thought to represent a cerebral insult caused by CPB, the magnitude of the insult might be less than previously assumed. PMID- 16352038 TI - Transcatheter embolisation of coronary artery fistulae. AB - BACKGROUND: Most children with coronary artery fistulae are asymptomatic, but because of associated late morbidity, early intervention is usually indicated. AIM: To assess the outcome following transcatheter embolisation of coronary artery fistulae. PATIENTS AND METHODS: Six children, with a median age of 9.5 years (range: 1.3-13.7 years), underwent transcatheter embolisation of coronary artery fistulae. Four patients had simple fistulae, which drained from the right coronary artery to the right ventricle (n = 2), the left coronary artery to the right ventricle (n = 1), or the left coronary artery to the coronary sinus (n = 1). Two patients had complex multiple fistulae arising from both coronary arteries, which communicated with either the right ventricle or the pulmonary arterial system. RESULTS: A stable position with a 5-Fr coronary catheter was obtained proximally and a 3-Fr coaxial catheter was advanced through the coronary catheter to a distal position in the coronary artery fistulae. The number of embolisation microcoils used per procedure ranged from one to 12, and the coil diameter ranged from 3 mm. Polyvinyl alcohol foam embolisation particles (1000 microm) were used to embolise small fistulae to the pulmonary arterial tree. Complete occlusion was obtained in four patients, while two children were left with insignificant residual shunts. There were no early or late cardiac complications. CONCLUSION: Transcatheter embolisation of coronary artery fistulae is a safe and effective therapy in patients with suitable anatomy. PMID- 16352039 TI - Intravascular ultrasound assessment of ambiguous coronary lesions. AB - Accurate assessment of coronary lesions is essential for clinical decision making. While angiography has long been accepted as the gold standard investigation, this technique provides only a planar 2-D silhouette of the arterial lumen and therefore has limited accuracy in the setting of vessel tortuosity or overlap, bifurcational and eccentric lesions, and diffusely diseased arteries. By providing high-resolution cross-sectional imaging through the arterial wall, intravascular ultrasound (IVUS) can overcome many of these limitations and accurately quantify angiographically indeterminate lesions. Angiographic evaluation of the left main coronary artery presents particular challenges that are ideally resolved with IVUS examination. The role of IVUS in the assessment of coronary stenoses of angiographically intermediate severity (50 70%) continues to evolve. Recent data correlating IVUS with intracoronary flow and pressure measurements suggest that epicardial coronary artery lesions with minimum lumen area of less than 3-4 mm2 may be haemodynamically significant. In addition to accurately quantifying minimum lumen diameter and area at the lesion site, IVUS can characterise coronary artery plaque morphology, and it may have the potential to predict plaque complications. PMID- 16352040 TI - Percutaneous transluminal septal myocardial ablation: a new catheter-based therapy for patients with hypertrophic obstructive cardiomyopathy. AB - Hypertrophic obstructive cardiomyopathy is a complex disorder with serious clinical implications. Percutaneous transluminal septal myocardial ablation is a promising new addition to existing therapies for this condition. It is a catheter based approach that involves instilling alcohol into the septal branches of the left anterior descending artery to induce a 'controlled' septal myocardial infarct. The result is a decrease in thickness of the hypertrophied interventricular septum and a reduction of the left ventricular outflow tract gradient. To date, the results from several series have been promising, with improvements in haemodynamic and clinical parameters without prohibitive complication rates. In this article, the indications, technique and outcomes of this procedure are reviewed. PMID- 16352041 TI - Anti-thrombotic, anti-platelet and fibrinolytic therapy: current management of acute myocardial infarction. AB - Significant advances in the treatment of patients with acute myocardial infarction (MI) have been obtained in recent times. In particular, thrombolytic therapy has been shown to preserve ventricular function and improve survival in patients with acute MI. Therapies now include third-generation thrombolytic agents, percutaneous transluminal coronary angioplasty (PTCA) and intracoronary stenting, and new anti-thrombotic therapies including anti-platelet treatment with glycoprotein (GP) IIb/IIIa inhibition and direct anti-thrombin agents. This review will focus on the use of GP IIb/IIIa antagonists and thrombin inhibitors as adjunctive therapies to thrombolytic treatment of patients with acute MI. PMID- 16352042 TI - Echocardiographic aspects of percutaneous atrial septal defect closure in adults. AB - Echocardiography plays an integral role in the diagnosis and management of atrial septal defects. With percutaneous closure of secundum defects becoming widespread, transoesophageal echocardiography in particular has assumed an important role in patient selection, as well as being central to the device deployment procedure, and an important tool in patient follow up. PMID- 16352043 TI - Does unprotected left main-stem stenting have a role in current interventional practice? AB - Left main-stem disease is found in up to 5% of patients undergoing coronary angiography. Until recently, it has been regarded as an absolute indication for coronary bypass surgery, given the poor results of the early trials of balloon angioplasty for this condition. However, with rapidly advancing technology, including new generation stents and increasingly effective post-stenting anti thrombotic regimens, there is now an increasing body of evidence to support the consideration of a percutaneous approach to left main-stem disease. Discerning patient selection, meticulous stent sizing and deployment, the routine use of intravascular ultrasound, aggressive anti-platelet regimes, and careful patient follow up are a few of the technical considerations required for a successful long-term outcome in this group. PMID- 16352044 TI - Balloon mitral valvuloplasty at Monash Medical Centre: follow up of 201 procedures over an 11-year period. AB - Over a period of 11 years from 1988 to 1999, 201 patients underwent balloon mitral valvuloplasty (BMV) at Monash Medical Centre, Australia. Before BMV,133 patients (66%) were symptomatic with minimal activity or at rest. BMV increased mitral valve area and cardiac output, and reduced transmitral, left atrial and pulmonary pressures, with infrequent procedural complications (<8%). At the initial 3-month follow up after BMV, symptoms were absent or minimal in 178 patients (89%), with 85% remaining event free at 12 months. At long-term follow up (median: 30 months; range: 0-129 months), cumulative event-free survival was 73% after 5 years. After BMV, 37 patients (18%) underwent mitral valve surgery, while a repeat BMV was performed in three patients (1.5%). The results of this series provide additional data for the growing body of evidence which suggests that BMV is a relatively safe and effective procedure for producing long-term benefit in patients with symptomatic mitral stenosis. PMID- 16352045 TI - Combination thrombolytic therapy and percutaneous coronary intervention: the future for revascularisation for acute myocardial infarction? AB - It seems theoretically sound to consider rescue percutaneous coronary intervention (PCI) in patients with failed thrombolysis. However, randomised controlled data in this setting are limited. In this brief review we will present those trials analysing the role of rescue PCI in patients with acute myocardial infarction and failed thrombolysis, and consider appropriate management strategies in this setting. PMID- 16352046 TI - Angiotensin II and cardiac excitation-contraction coupling: questions and controversies. AB - Angiotensin II (AngII) is a circulating peptide that produces a positive inotropic effect in the heart in several species, including humans. The subcellular mechanisms involved in producing this effect have been the focus of numerous studies; however, the results of these studies have generated considerable controversy. Although part of the controversy might arise from species and developmental differences, conflicting results have also been reported in the same species. To further complicate the understanding of the cardiac actions of AngII, the binding of the peptide to its transmembrane G protein-coupled receptors has been shown to activate signalling cascades that involve numerous second messengers. Among these, inositol 1,4,5-triphosphate (IP3) and protein kinase C (PKC) have been shown to have the potential to modulate either one or both of the two basic mechanisms known to increase contractility: (i) an increase in the intracellular Ca2+ concentration ([Ca2+]i); or (ii) an increase in myofilament responsiveness to Ca2+. The aim of this review is to examine the effect of AngII on the fundamental components of cardiac excitation-contraction coupling: calcium currents, Na+/Ca2+ exchange, sarcoplasmic reticulum (SR)-CaZ+ release, calcium transients and contractile proteins. An answer to the following question is sought: Is the positive inotropic effect of AngII due to an increase in [Ca2+]i, to an increase in myofilament responsiveness to Ca2+, or to both? PMID- 16352047 TI - Papillary fibroelastoma, a rare but potentially treatable cause of embolic stroke: report of three cases. AB - Papillary fibroelastomas are rare benign tumours that are seldom diagnosed during life. We report three cases of this tumour; two patients had recurrent embolic events, and one was diagnosed incidentally. Although rare, these tumours are a potentially treatable cause of cardiac emboli. Prompt identification allows excision, which generally is curative. PMID- 16352048 TI - The other second messenger gas: carbon monoxide signalling for myocardial protection? PMID- 16352049 TI - Coffee and coronary heart disease. AB - BACKGROUND: The role of coffee consumption in the onset of myocardial infarction remains uncertain. A review of published reports showed that although cohort data suggest very little excess risk of coronary heart disease among habitual coffee drinkers, case-control data suggest an excess risk of the order of 60% for people drinking five or more cups per day. METHODS: We obtained as much information as possible on the lifestyle and habits, including coffee consumption, of people admitted with chest pain to the Coronary Care Unit at the Royal Perth Hospital, Australia. A questionnaire was given to them by the attending nursing staff. Details were recorded by the patient, often with help from the nursing staff. A similar questionnaire was completed by patients attending the Cardiology Outpatient Clinic. RESULTS: One of the outstanding differences between the 182 patients with myocardial infarction or unstable angina and 185 patients with chronic stable coronary heart disease who filled in forms while waiting in the Cardiology Outpatient Clinic was that more people with acute coronary syndromes were drinking in excess of five cups of coffee per day (18 vs 7.5%; P = 0.003). In logistic regression analysis adjusted for age, smoking status (past and current), hypertension, dyslipidaemia and diabetes, the odds ratio was 2.51 (95% CI 1.43,4.43; P = 0.021). CONCLUSION: The present case-control study provides evidence for an increased risk of myocardial infarction or unstable angina among individuals drinking more than five cups of coffee per day. PMID- 16352050 TI - Hypoxia-induced Haem Oxygenase-1 gene expression in neonatal rat cardiac myocytes. AB - BACKGROUND: Haem oxygenase (HO-1), a heat shock or stress protein, is a rate limiting enzyme in the conversion of pro-oxidant haem to biliverdin and carbon monoxide (CO). The products of haem catabolism serve regulatory and protective functions. Previous studies have shown that hypoxia induces HO-1 expression in cardiac myocytes. Accordingly, we investigated whether hypoxia-induced HO-1 expression is accompanied by increased CO production in cultured neonatal rat cardiac myocytes, and whether protein kinase C (PKC) is involved in hypoxia induced HO-1 gene expression. METHODS AND RESULTS: Expression of HO-1 in hypoxia treated cells was examined by using northern and western blotting, and immunofluorescent staining. The level of HO-1 mRNA at 24 and 48 h was increased after the onset of hypoxia, with corresponding increase in the HO-1 protein level (6.7- and 8.7-fold at 24 and 48 h of hypoxia, respectively). HO-1 protein was colocalised with sarcomeric alpha-actin in hypoxic myocytes. Hypoxia also significantly increased the production of CO by 2.5- and 8-fold at 24 and 48 h, respectively. Under normoxic conditions, activation of PKC by phorbol-12 myristate-13-acetate (PMA; 100 nmol/L) markedly increased HO-1 gene expression, while inhibition of PKC activity by calphostin C (100 nmol/L) blocked hypoxia induced HO-1 gene expression in cardiac myocytes. CONCLUSIONS: These results demonstrate that hypoxia markedly induces HO-1 expression and increases the production of CO in cardiac myocytes. This hypoxic response is attributed, at least in part, to activation of PKC. Increased HO-1 expression and resultant CO production may be beneficial with respect to protection of cardiac myocytes under hypoxic conditions. PMID- 16352051 TI - Subaortic membrane excision: mid-term results. AB - BACKGROUND: Subaortic membrane (SAM) is a form of fixed subaortic obstruction in which a fibrous membrane is located below the aortic valve. AIM: To determine the role of surgical treatment for patients with a discrete SAM. PATIENTS AND METHODS: The hospital records of 45 patients (age range: 2-23 years; median 8 years) undergoing surgery for SAM between 1990 and 1998 at the All India Institute of Medical Sciences, New Delhi, India, were analysed. Preoperative echocardiographically calculated gradients across the left ventricular outflow tract ranged from 50 to 154 mmHg (mean: 86.5 +/- 33.2 mmHg). Nine patients had trivial aortic regurgitation (AR), 10 had mild AR and five had moderate-severe AR. The left ventricular ejection fraction (LVEF) ranged from 20 to 68% (mean 48 +/- 15%). Nineteen patients had significant left ventricular dysfunction (LVEF <50%). Transaortic resection of SAM was done in all patients along with excision of a wedge-shaped segment of septal muscle underlying the membrane. RESULTS: There were no early or late postoperative deaths. On follow up (up to 113 months), only four patients had gradients >30 mmHg. LVEF improved to 45-70% (mean 58 +/- 7.7%). AR reduced to mild in four patients and trivial in four patients, and did not progress further. CONCLUSION: Resection of SAM carries long-term benefits. Routine septal myectomy appears to be associated with a low risk of recurrence. PMID- 16352053 TI - Practical management of anticoagulation, bleeding and blood product support for cardiac surgery. Part one: bleeding and anticoagulation issues. AB - There are many challenging problems related to bleeding and anticoagulation in cardiac surgery. Practical guidelines, which are based on available evidence, can help to direct management issues of post-bypass bleeding, perioperative anticoagulation and the use of haemostatic agents. The patient's bleeding history is the most useful preoperative screening test of haemostasis. The input of a haematologist is often valuable in a number of areas, such as preoperative assessment of patients with a significant history of bleeding, or past history of heparin-induced thrombocytopenia, a lupus anticoagulant, or recent venous thromboembolism. Cardiothoracic surgeons, anaesthetists, perfusionists and haematologists can ensure 'best practice' by being actively involved in the development of 'local' transfusion and anticoagulation guidelines, hospital transfusion committee policies, and audits. PMID- 16352052 TI - Stage one Norwood procedure results: the green lane experience. AB - AIMS: This study examined the presentation outcome, morbidity and mortality of infants who have undergone the stage one Norwood procedure for single ventricle reconstruction. METHODS: A retrospective review was done on the first 20 patients to undergo this procedure at Green Lane Hospital, Auckland, New Zealand. Seven patients were diagnosed antenatally. Fetal cardiology records in the same time period were reviewed. RESULTS: Twelve of the 20 patients (60%) have survived, and all of these patients have undergone their bi-directional Glenn procedure with no mortality. Eight patients died, with five of the deaths occurring in the perioperative period. Initial surgical mortality was 75%, decreasing to 25% since 1998. Antenatal diagnosis has not improved surgical outcome to date. CONCLUSION: With advances in surgical technique and pre- and postoperative care, neonates born with single ventricle anatomy have an acceptable surgical option. Babies who survive the Norwood operation have a good chance of surviving the later stages of the cardiac reconstruction process, and they have a reasonable outlook in the intermediate term. PMID- 16352054 TI - Aneurysms of an aberrant right subclavian artery: report of two cases. AB - Aneurysm of the aberrant right subclavian artery is rare. We report two patients who underwent successful repair using hypothermic circulatory arrest and retrograde cerebral perfusion. One patient presented with progressive dysphagia to solids and hoarseness of voice due to pressure exerted by the expanding aneurysm, and the second patient, with a dissected subclavian aneurysm, presented with transient ischaemic attacks and a previous cerebral infarct secondary to embolism. Repair in both patients was done through a median sternotomy with femoral arterial and right atrial venous cannulation. Under hypothermic circulatory arrest, the origin of the subclavian aneurysm was divided off the aorta and closed with a Dacron patch from outside the aorta in one patient, and from within the aorta in the other patient. Concomitant revascularisation of the right upper limb was achieved using a 12-mm Hemishield Dacron graft from the ascending aorta to the distal right subclavian artery. The graft to subclavian anastomosis was done under hypothermic circulatory arrest as distal control was not possible. The graft to the aortic anastomosis was done over a side-biting clamp during rewarming. Both patients were successfully treated without major morbidity. PMID- 16352055 TI - An unusual case of aortic dissection in Turner's syndrome. AB - Cardiovascular malformations are common in patients with Turner's syndrome. Aortic coarctation and bicuspid aortic valve are the most frequently occurring abnormalities, and are associated with cystic medial necrosis of the aortic wall. Aortic dissection is an uncommon but catastrophic complication of the 'aortopathy' of Turner's syndrome. We report the unusual case of a Turner's syndrome patient (with a bicuspid aortic valve and previous coarctation repair) who died following an intramural haemorrhage of the aortic root that was complicated by dissection and rupture, with no evidence of aortic intimal tear. The role of intramural haemorrhage in the pathogenesis of acute aortic syndromes in Turner's syndrome patients is unclear. The condition may be associated with atypical clinical presentations, it can be difficult to confirm with imaging techniques, and it carries a high risk of progression to classical aortic dissection and death. This case therefore highlights the need for a high index of suspicion when assessing Turner's syndrome patients presenting with chest pain syndromes. Furthermore, the effective management of Turner's syndrome patients with cardiovascular abnormalities requires the development of evidence-based preventive (such as echocardiographic surveillance of aortic dilatation) and interventional strategies. PMID- 16352056 TI - Mural endocarditis caused by Salmonella virchow: survival with conservative treatment. AB - We describe a case of endocarditis caused by Salmonella enterica serotype virchow, which was treated conservatively with antibiotics alone. It is the only reported case of survival from salmonella endocarditis with conservative treatment, and the first reported case of endocarditis caused by Salmonella virchow. The changing prevalence, virulence patterns and importance of salmonella species in endocarditis are discussed. PMID- 16352057 TI - Cardiac angiosarcoma: too little known, too late treatment or just too bad a tumour? Heart, Lung and Circulation 2001; 10: 30-34). PMID- 16352059 TI - Relevancy of the National Heart Foundation. PMID- 16352060 TI - The cardiothoracic surgery workforce in Australia: new approaches needed for a changing profession? PMID- 16352062 TI - Anticoagulation and transfusion management in cardiac surgery. PMID- 16352061 TI - Cardiac rehabilitation after acute myocardial infarction: for the many or the few? PMID- 16352063 TI - Economic evaluation of a randomised trial of early return to normal activities versus cardiac rehabilitation after acute myocardial infarction. AB - BACKGROUND: Although there have been a number of economic evaluations of cardiac rehabilitation after acute myocardial infarction (AMI), none has considered only low-risk patients or control groups with no rehabilitation at all. METHODS: An economic evaluation was included in a randomised controlled trial of patients following uncomplicated AMI. Eligible patients were randomised to return to normal activities after 6 weeks of standard rehabilitation (REHAB, n = 70) or to early return to normal activities 2 weeks after AMI with no formal rehabilitation (ERNA, n = 72). Outcomes were assessed weekly for 6 weeks, then 3, 6 and 12 months post-AMI. Outcomes included four quality of life (QOL) measures (physical abilities, distress, usual/social activities, self-care) and four measures of return to normal activities (paid and unpaid return to any work and to pre-AMI level of work). Statistical analysis included repeated-measures regression (QOL outcomes) and survival analysis (work outcomes). RESULTS: There were no statistically significant differences between the two groups in any of the outcomes measured or in the use of other health services. The net cost that could be saved by the health service by targeting rehabilitation to high-risk patients was approximately $300 (Australian, 1999) per low-risk patient. CONCLUSIONS: Early return to normal activities without formal rehabilitation is cost-effective for low-risk patients. PMID- 16352064 TI - Coronary artery imaging in the new millennium. AB - Atherosclerotic disease and its thrombotic complications remain the leading causes of mortality and morbidity in Western society. In Australia, cardiovascular disease is responsible for one in every 2.4 (41%) deaths and is the leading single cause of mortality. The crucial final common process for the conversion of a non-occlusive, often clinically silent, atherosclerotic lesion to a potentially fatal condition is plaque disruption. The mortality associated with atherosclerotic disease relates to the acute coronary syndromes, including acute myocardial infarction, unstable angina pectoris and sudden cardiac death. There is substantial clinical, experimental and postmortem evidence demonstrating the role acute thrombosis upon a disrupted atherosclerotic plaque plays in the onset of acute coronary syndromes. Atherosclerotic plaque composition, rather than the stenotic severity, appears to be central in determining risk of both plaque rupture and subsequent thrombogenicity. In particular, a large lipid core and a thin fibrous cap render an atherosclerotic lesion susceptible or vulnerable to these complications. We are currently limited in our ability to accurately identify patients at risk for an acute coronary event. The armamentarium of diagnostic investigations, both non-invasive and invasive, currently clinically available is only able to provide us with data related to the stenotic severity of a coronary artery. The non-invasive testing includes stress-induced (exercise or pharmacological) ischaemic changes in electrical repolarisation, wall motion or myocardial radioactive-tracer uptake. The invasive test of coronary angiography, although the current 'gold standard' for the detection of coronary atherosclerotic disease, provides us with no data about the composition of the atherosclerotic lesion. However, the vast majority of acute coronary events involve a non-critically stenosed atherosclerotic lesion, and thus with currently available means of identification, these lesions would be undetected by stress testing/imaging techniques. Given the critical role that atherosclerotic lesion composition has been shown to play in the risk of both plaque rupture and subsequent thrombogenicity and, consequently, an acute coronary event, new detection techniques need to be investigated for the task of documenting atherosclerotic lesion composition. In the present review we will focus on the status of imaging modalities available for coronary artery imaging and how they may advance our understanding and management of patients with and at risk of coronary artery disease in the new millennium. PMID- 16352065 TI - The changing profile of bacterial endocarditis as seen at an australian provincial centre. AB - BACKGROUND: The clinical profile of endocarditis has changed over the past four decades with studies showing trends towards increasing age, more nosocomial and prosthetic valve infection and increasing rates of Staphylococcus aureus infection. However, these studies have been biased by referral patterns. METHODS: We reviewed data collected at three hospitals in the Barwon-South-West region in Victoria, Australia. All cases identified between 1994 and 1999 were reviewed according to the Duke criteria. RESULTS: During this period, 58 patients were diagnosed as having endocarditis. The incidence rate during this time was 3.0 per 100 000 with a rise in the rate of admissions from 0.15 to 0.26 per 1000 from 1995 to 1999. Sixteen (28%) were nosocomial with the majority from line-related sepsis. No intravenous drug users were identified. Eighteen (31%) involved endovascular prosthetic material. S. aureus was the causative pathogen in 23 (40%), with 'viridans' streptococci contributing 12 (21%) and other organisms accounting for 12 (21%). Attributable mortality in this series was 17%. CONCLUSIONS: We have seen a rise in the rate of endocarditis during this time. The proportions of endocarditis due to S. aureus and 'viridans' streptococci, as well as rates of nosocomial and prosthetic valve infection, are consistent with more recent series at referral hospitals and district hospitals, representing a change since studies performed in the 1960s and 1970s. Our findings confirm a trend towards a clinical profile seen at referral centres and reinforce the emerging importance of S. aureus, nosocomial bacteraemia and prosthetic valve endocarditis. PMID- 16352066 TI - Xenotransplantation: past achievements and future promise. AB - Xenotransplantation offers a potential solution to the shortfall in donor organs for human transplantation. This review describes the barriers to xenotransplantation and the progress that has been made towards making it a clinical reality. Data from preclinical pig-to-primate cardiac and pulmonary xenografts are highlighted. PMID- 16352067 TI - Practical management of anticoagulation, bleeding and blood product support for cardiac surgery. Part two: transfusion issues. AB - We summarise recent advances in transfusion medicine applicable to cardiac surgery and cardiac transplantation. It is important that clinicians know the risks of blood transfusion in Australia. They should also be aware of the different types of transfusion reaction so that there is early recognition and investigation. Blood conservation strategies including acceptance of normovolaemic anaemia in clinically stable patients are important in reducing the requirement for red cell transfusion. Cytomegalovirus (CMV) seronegative blood products are recommended for heart transplant recipients with no evidence of prior CMV infection. Leucodepletion of units of unknown CMV status reduces the risk of CMV infection and are an acceptable alternative when seronegative units are unavailable. Leucodepletion of cellular blood products has been shown to reduce infection rates postoperatively in a large trial involving cardiac surgical patients. Further studies are needed to confirm this promising finding. Irradiation of blood products eliminates the risk of transfusion-associated graft versus host disease. Routine preoperative screening for cold agglutinins is no longer recommended. PMID- 16352068 TI - The cardiothoracic surgery workforce in Australia: supply and requirements 2000 2011. Report of the Australian Medical Workforce Advisory Committee 2001. AB - To establish a national profile of the current cardiothoracic surgery workforce and advise the Australian Government on workforce requirements to 2011, the Cardiothoracic Surgery Workforce Working Party brought down a report in May 2001 to The Australian Medical Workforce Advisory Committee. The survey data were taken from half the current Fellows of the Royal Australian College of Surgeons engaged in cardiothoracic surgery. Australian cardiothoracic surgeons are typically middle-aged males working more than 60 h per week. Their work has both private and public sector components and most surgeons provide services in both cardiac and thoracic surgery to an average catchment of 180 000 people. Concentrated in the capital cities, mainly on the eastern seaboard, they devote 90% of their working hours to service delivery, with little time spent in research and developmental aspects of their profession, or in administrative roles. The committee provided 10-year projections for surgical services, predicting an annual growth in service requirements of 1.8% in Australia to 2011. The report concludes that maintenance of the present intake of surgical trainees at five per year - offset by the projected retirements of 2.7 surgeons annually - will be sufficient to meet demand. This conclusion is based on an assumption of a decade of unchanged national health structures and patterns of workforce participation and service delivery. PMID- 16352069 TI - Amiodarone pneumonitis diagnosed by gallium-67 scintigraphy. AB - A 75-year-old man presented with fever, dyspnoea, chest pain and findings suggestive of severe pneumonia including hypoxia and bilateral chest infiltrates. He was given antibiotics without significant effect. He took long-term amiodarone therapy for persistent atrial arrhythmias and also had a history of asbestosis with pre-existing pulmonary interstitial fibrosis. Pulmonary gallium-67 scintigraphy revealed extensive uptake of tracer throughout both lungs consistent with a diagnosis of amiodarone pneumonitis. Treatment of the drug reaction by drug withdrawal and corticosteroids resulted in marked clinical improvement. Pulmonary gallium-67 scintigraphy is a sensitive, but non-specific aid in the diagnosis of this drug effect. It proved valuable in our patient who additionally had asbestosis. PMID- 16352070 TI - Utility of cardiovascular magnetic resonance imaging. PMID- 16352071 TI - Coronary artery bypass graft: on- or off-pump? PMID- 16352072 TI - The ageing heart: a clinical cardiologist's perspective car. PMID- 16352073 TI - The Australasian section of the International Society for Heart Research. PMID- 16352074 TI - Cardiovascular ageing in health sets the stage for cardiovascular disease. AB - The incidence and prevalence of coronary disease, hypertension, heart failure and stroke increase exponentially with advancing age. While epidemiologic studies have discovered that aspects of lifestyle and genetics are risk factors for these diseases, age, per se, confers the major risk. Thus, it is reasonable to hypothesise that specific pathophysiological mechanisms that underlie these diseases become superimposed on cardiac and vascular substrates that have been modified by an 'ageing process', and that the latter modulates disease occurrence and severity. In order to unravel this age-disease interaction, the nature of the ageing process in the heart and vasculature requires elucidation. Some aspects of the current understanding of ageing of the heart and blood vessels in the absence of apparent disease are the focus of this review. PMID- 16352075 TI - Tissue doppler imaging in hypertrophic cardiomyopathy without left ventricular hypertrophy. AB - BACKGROUND: Diastolic function abnormalities determined by Doppler echo (especially those recorded by tissue velocity imaging) may be useful in determining carriers of an abnormal gene in hypertrophic cardiomyopathy who do not yet show clinical evidence of the disease. METHODS: In a single extended family, seven carriers of a mutation involving the cardiac myosin-binding protein C gene who did not show any features of the disease on 2-D echocardiography were examined by 2-D, M-mode Doppler and tissue Doppler imaging. The results were compared with a group of eight aged-matched people from the same family who did not have the gene mutation or clinical evidence of the disease. In both groups, as well as measuring the basal posterior and anteroseptal walls, the entire left ventricle was inspected for localised hypertrophy. RESULTS: Localised hypertrophy was absent from the left ventricle in both groups. There were significant differences in the tissue Doppler peak velocity measurements made at the lateral border of the mitral annulus in systole and especially early diastole. A systolic velocity of <10 cm/s was strongly suggestive of gene positivity and, if combined with an early mitral diastolic velocity (mitral E velocity) of <14 cm/s, was present only in gene-positive individuals. If the mitral E velocity was >14 cm/s, this was present only in gene-negative patients. CONCLUSION: Tissue Doppler imaging may be a valuable tool for screening first-degree relatives of patients with hypertrophic cardiomyopathy who do not show 2-D echo evidence of the disease. PMID- 16352076 TI - Neuropsychological and psychiatric outcomes following coronary surgery or angioplasty: a comparative study. AB - BACKGROUND: Medical outcomes following coronary artery bypass grafting (CABG) or percutaneous transluminal coronary angioplasty (PTCA) are similar, but few studies have compared neuropsychological outcomes after these procedures. METHODS: A retrospective study compared detailed neurocognitive and psychosocial functioning in 32 patients (CABG, n = 16; PTCA, n = 16) aged 61 +/- 6 years, 9-15 months after coronary revascularisation. Subjects were tested for executive functioning, speed of processing/attention and learning/memory, significant psychopathology (General Health Questionnaire, GHQ) and psychosocial functioning (Short Form (SF)-36 health survey). In the prospective study, 55 patients completed GHQ and SF-36 surveys, the day prior to and 6 months following PTCA. RESULTS: There were no significant differences between the CABG and PTCA groups for neuropsychological or psychosocial end-points (P > 0.20). Executive functioning in both groups, however, was worse than for healthy population controls (P < 0.01). The PTCA patients were significantly more likely than CABG patients to have psychiatric abnormality (GHQ Score >4; P < 0.01). After PTCA, however, there was a significant improvement in the GHQ and SF-36 scores (P < 0.05). CONCLUSIONS: Although executive function is often impaired after coronary revascularisation, neuropsychological status appears equivalent after CABG or PTCA. Psychiatric pathology is common in patients undergoing PTCA, but improves after this intervention. PMID- 16352077 TI - No decrease in incidence of atrial fibrillation in patients undergoing off-pump coronary artery bypass grafting. AB - BACKGROUND: With the introduction of beating heart surgery, there has been a renewed interest in the incidence of postoperative atrial fibrillation (AF) in patients undergoing coronary artery bypass grafting (CABG). A small number of studies have reported markedly decreased incidences of postoperative AF in patients undergoing off-pump CABG. AIMS: The aim of the present study was to review the incidence of AF in patients undergoing off-pump CABG at Royal Melbourne Hospital. METHODS: A retrospective review was undertaken of 80 patients who had elective first-time CABG off-pump over a 16-month period. These patients were matched with 80 patients who underwent CABG with cardiopulmonary bypass (CPB). Data on preoperative and operative variables were collected. Incidences of postoperative AF in both groups were recorded. RESULTS: The incidence of AF in the off-pump group was 27 of 80 patients (34%) and in the CPB group it was 25 of 80 patients (31%; NS). Increasing age was a significant risk factor for developing postoperative AF, and beta-blockade was identified as a protective factor. CONCLUSIONS: Off-pump CABG does not decrease the incidence of postoperative atrial fibrillation. A randomised prospective trial has been commenced at Royal Melbourne Hospital to further investigate postoperative outcomes of off-pump CABG. PMID- 16352078 TI - Treatment and outcome of sinus of valsalva aneurysm. AB - BACKGROUND: In order to understand the long-term outcome after surgical repair of sinus of Valsalva aneurysms or fistulas (SVAF), the operative results for patients treated at Shanghai Chest Hospital were retrospectively assessed. METHODS: Between 1960 and 1999, 216 patients were operated on for SVAF. Of these 143 (66.2%) had a ventricular septal defect (VSD), 60 (28.0%) had aortic regurgitation and of these 12 underwent aortic valve replacement (AVR). RESULTS: There were eight in-hospital deaths (3.7%). Another patient had aortic regurgitation postoperatively requiring AVR. CONCLUSION: Resection and repair of SVAF entails an acceptably low operative risk and yields long-term freedom from symptoms. Early, aggressive operation is recommended to prevent development of complications. PMID- 16352079 TI - Antimalarial drugs inhibit the acetylcholine-receptor-operated potassium current in atrial myocytes. AB - BACKGROUND: It has been reported that halofantrine, an antimalarial drug, was associated with electrocardiographic prolongation of the QT interval and ventricular arrhythmias. Inhibition of the delayed rectifier potassium channel, a voltage-gated potassium channel, by halofantrine was the likely underlying cellular mechanism for this cardiotoxicity. However, influences of anti-malarial drugs on the ligand-gated potassium channels have not been well-documented. The influences of three different antimalarial drugs, chloroquine, primaquine and pyrimethamine, on the acetylcholine-receptor-operated potassium current (I(K.ACh)), a ligand-gated potassium current, were compared with the effect of quinidine in isolated guinea pig atrial myocytes using patch-clamp techniques. METHODS: The whole-cell patch-clamp method was used in the present studies he I(K.ACh) was induced by extracellular application of carbachol (1 micromol/L) or intracellular loading of guanosine 5'-O-(3-thiotriphosphate) GTPgammaS (100 micromol/L) in acutely isolated guinea pig atrial myocytes. RESULTS: The I(K.ACh) induced by carbachol was inhibited by chloroquine, primaquine, pyrimethamine and quinidine in a concentration-dependent manner, and the concentrations required to produce 50% of the maximal inhibitory effect (IC(50) values) were 0.7, 2.5, 12 and 1.8 micromol/L, respectively. These drugs also inhibited the intracellular GTPgammaS-activated I(K.ACh), and the IC(50) values were 0.8,13,19 and 21 micromol/L, respectively. CONCLUSIONS: Chloroquine and pyrimethamine may inhibit I(K.ACh) by interacting with the muscarinic potassium channel itself and/or associated guanosine 5'-triphosphate-binding proteins, whereas primaquine and quinidine may mainly inhibit the current by the blockade of the muscarinic receptors. These results indicate that antimalarial drugs exert anticholinergic effects via different molecular mechanisms. PMID- 16352080 TI - Cardiac hydatid cyst: report of two cases. AB - Cardiac hydatid cyst is life threatening but rare. It is usually asymptomatic and has potentially lethal complications, so early diagnosis with definitive treatment is life-saving. In the present article, we report two cases. The first is a 30-year-old woman with a primary large multivesicular hydatid cyst in the left ventricle. The second is a 17-year-old woman who had multivisceral involvement with a cardiac hydatid cyst, a congenital cardiac anomaly that was an atrial septal defect complicated by pulmonary hypertension. 2-D echocardiography and magnetic resonance imaging confirmed the diagnosis. Surgical treatment was performed using extracorporeal circulation and adjunctive albendazol therapy. PMID- 16352081 TI - Alternative approach for angioplasty of stenosed left pulmonary artery following intracardiac repair of tetralogy of Fallot. AB - A 5-year-old boy with stenosed left pulmonary artery following total correction of tetralogy of Fallot underwent surgical pulmonary arterioplasty through a left anterolateral thoracotomy on a normothermic perfused heart under cardiopulmonary bypass. We found this to be a convenient approach, and recommend it for correction of this lesion in the absence of gross pulmonary regurgitation and right ventricular outflow tract dilatation. PMID- 16352082 TI - Functional tricuspid regurgitation secondary to aortic annular abscess: An unusual presentation of infective endocarditis. AB - An unusual case of aortic annular abscess is presented, in which the patient presented with features of gross tricuspid regurgitation. There was no direct involvement of the tricuspid valve. Tricuspid regurgitation disappeared following surgical repair of the annular abscess. The present case also illustrates the utility of trans-oesophageal echocardiography in establishing the diagnosis and planning surgical intervention. PMID- 16352083 TI - Will tissue engineering become the 21st century's answer to cardiovascular disease? PMID- 16352084 TI - Progress in the treatment of in-stent restenosis: role of radiation therapy and its use in Australia. PMID- 16352085 TI - Early extubation after paediatric cardiac surgery. PMID- 16352087 TI - Tissue engineering of biological cardiovascular system surrogates. AB - Cardiovascular diseases are common in ageing communities globally. This fact is most striking in the industrialised world where the aged population makes up a large proportion of society. Elderly patients are frequently treated surgically with grafts to replace damaged tissues and vessels. The number of human-donated components is insufficient and synthetic surrogates are sought. These might be wholly mechanical, wholly biological, or tissue engineered complexes of cells and their products growing in a scaffold. At present, many such composites exist with potential for use as substitutes for specific blood vessels. The challenges of producing tissue engineered heart valves are now being widely explored. Neotissues must provide an effective, durable, non-thrombogenic and non immunogenic substitute that will fulfil the purpose of the natural tissue. The aims and scope of this paper are to review current and novel concepts in the field of tissue engineering of biological cardiovascular system surrogates. Mechanical stresses and strains on cardiovascular cells in vitro have been recognised and can be measured by a culture force monitor. Physiological stresses can be generated by a tensioning culture force monitor and applied to engineered tissue, aligning the cells and mimicking arterial wall architecture. The hydrostatic forces a vessel experiences and mechanical parameters of blood vessels can be studied in the tubular culture system of a multi-cue bioreactor. PMID- 16352089 TI - Blalock-Taussig shunt: experience from the developing world. AB - BACKGROUND: Palliative procedures have a role in congenital cardiac malformations that do not permit a complete early repair, and in centres where facilities for complete early repair do not exist. The lack of data on modified Blalock-Taussig shunt from developing countries prompted this analysis. METHODS: We report a retrospective study of 70 Blalock-Taussig shunt procedures in 63 patients over an 8-year period. Most of the procedures (54.0%) were done on children less than 4 months of age. Thirty-nine (58%) patients had Tetralogy of Fallot; the remaining patients had a wide spectrum of lesions. RESULTS: In the first year of the review period, the classical Blalock-Taussig shunt was done in six patients (9.5%) and the modified Blalock-Taussig shunt was used thereafter. In 49 patients who were followed up long term, clinical congestive cardiac failure developed in three (6%) and shunt failure was reported in 10 (14%). Of the 16 (33%) total deaths, six had serious comorbid conditions preoperatively. CONCLUSIONS: The Blalock Taussig shunt is a relatively safe palliative procedure, requiring fewer resources and less expertise than corrective surgery, making it a suitable option in developing countries. PMID- 16352090 TI - The cardiothoracic surgery workforce in Australia. PMID- 16352091 TI - Early extubation in congenital heart surgery. AB - BACKGROUND: The concept of early extubation following congenital heart surgery is not new. Changes in health care have generated increasing interest in this technique. Some of the problems following surgery are related to the endotracheal tube and mechanical ventilation, and the interventions necessary to maintain them. METHODS: To evaluate the impact of early extubation (within 6 h postoperatively) on children undergoing congenital heart surgery, retrospective data were obtained from records of 1000 consecutive patients. RESULTS: Early extubation occurred in 80.2% of patients (73% in the operating room). Early extubation was not limited to simple anomalies; it was also possible for complex anomalies such as Fallot's tetralogy (including those patients with pulmonary atresia, absent pulmonary valve and complete atrioventricular septal defects; n =106), where 91% of patients had early extubation, and Fontan operations (n =17), where 88% did. There were no deaths related to early extubation. Preoperative intubation was a risk factor for postoperative ventilation. As expected, the patients requiring ventilation after surgery were younger, smaller and more critically ill than those that met the criteria for early extubation. CONCLUSIONS: A change in attitude combined with appropriate anaesthetic and surgical techniques permitted safe, early extubation in a large number of patients. Patient populations vary between institutions. Early extubation is not always possible, but for those patients in whom it is feasible, the benefits include simplified postoperative care and increased patient and family satisfaction. When combined with clinical practice guidelines, it can result in a significant reduction in the cost of patient care after cardiac surgery. PMID- 16352092 TI - Intracoronary brachytherapy: Initial Australasian experience with the Novoste Beta-Cath system. AB - BACKGROUND: Coronary artery stenting is now performed in the vast majority of percutaneous interventional procedures worldwide. In-stent restenosis remains the major limitation of this technology and responds poorly to conventional interventional therapies. The only available treatment shown to impact upon this unfavourable response is endovascular radiation. In this paper we describe the initial Australasian experience with the Novoste Beta-Cath device, designed specifically for the delivery of beta radiation to the coronary artery. METHODS: Patients were considered eligible for intracoronary brachytherapy if they had symptomatic diffuse in-stent restenosis, or restenosis within a vessel that was not suitable for stenting. Brachytherapy was provided as a clinical procedure following successful percutaneous transterminal coronary angioplasty (PTCA). Clinical and procedural details were collected at hospital admission, and patients were contacted a median of 9 months following the procedure. Information was collected on recurrent hospital presentations and details verified by retrieving patient records. RESULTS: From June 1999 to July 2000, intracoronary brachytherapy was performed in 17 vessels in 16 patients and was technically successful in all cases. A single late stent thrombosis occurred 80 days following brachytherapy. This patient died of a subarachnoid haemorrhage 186 days later. There were three presentations with chest pain during the follow-up period, one associated with scintigraphic abnormality consistent with clinical restenosis. There was no documented angiographic restenosis. CONCLUSIONS: Intracoronary brachytherapy with the Beta-Cath device is simple to apply and has yielded promising intermediate term results in our patient cohort. Its application appears most appropriate to diffuse in-stent restenotic lesions in vessels ranging from 2.5 to 3.5 mm in diameter. In these patients, adjunctive brachytherapy currently provides the best available option for ensuring an enduring result following repeat percutaneous revascularisation. PMID- 16352093 TI - Echocardiographic assessment of cardiac structure and function in rats. AB - BACKGROUND: Echocardiography is used in humans to characterise the structure and function of the heart, yet is relatively uncommon in studies on the rat, the most commonly used model of human cardiovascular disease. The aim of this study was to show that echocardiography in rats provides useful information on cardiac changes occurring in thyroid dysfunction and can also be used to characterise cardiac abnormalities. METHODS: Transthoracic echocardiography and Doppler techniques with high frequency, high frame rate imaging were used to define cardiac dimensions and function in 240 Wistar rats and cardiac abnormalities in Wistar and spontaneously hypertensive rats (SHR). RESULTS: Echocardiographic assessment of left ventricular dimensions and function and aortic flows was technically feasible in almost all adult Wistar rats and SHR, including those with thyroid dysfunction and cardiac abnormalities. Pulsed-wave Doppler profiles of mitral inflows to estimate diastolic function were less reliably obtained. CONCLUSIONS: Echocardiography is a powerful technique for non-invasive and serial determination of cardiac structure and function in rat models of human cardiovascular disease. PMID- 16352094 TI - Gene therapy prevents disruption of dystrophin-related proteins in a model of hereditary dilated cardiomyopathy in hamsters. AB - BACKGROUND: The TO-2 hamster is an animal model of dilated cardiomyopathy (DCM). It has genetic and clinical features in common with humans who carry the gene deletion or mutation of the delta-sarcoglycan (SG) gene, a component in dystrophin-related proteins (DRP). DRP stabilise the sarcolemma during cardiac contraction. We performed in vivo gene therapy of the TO-2 hamster, whose heart is defective in all four SG proteins, to determine its potential as a model for therapy for DCM. In addition to the hereditary origin, heart failure is aggravated by treatment with catecholamines and ameliorated by the administration of some kinds of beta-antagonist both in humans and in TO-2 hamsters. METHODS: Gene therapy for DCM was achieved by supplementing the delta-SG gene with rAAV vector and intramurally delivering rAAV-delta-SG into the cardiac apex and left ventricle. RESULTS: This treatment resulted in: (i) a sustained and non pathogenic expression of both the transcript and transgene of delta-SG and all other SG proteins; (ii) improvement to both morphological and physiological deterioration; and (iii) rescued prognosis compared with untreated TO-2 hamsters, and TO-2 hamsters transfected with reporter gene alone. Another acute heart failure model was prepared by high-dose isoproterenol treatment in Wistar rats, which resulted in: (i) translocation of dystrophin, but not delta-SG, from the cardiac sarcolemma to the myoplasm; and (ii) fragmentation of dystrophin, probably due to the activation of endogenous protease(s) or proteasome(s) that contributed to muscular dystrophy-like degeneration occurring specifically in cardiomyocytes. CONCLUSIONS: Both the TO-2 hamster and the isoproterenol-treated Wistar rat models commonly experience disruption of dystrophin or DRP. Targeting the responsible gene with the use of a potent vector may provide a novel strategy for the treatment of advanced heart failure. PMID- 16352095 TI - New mouse model of vein bypass graft atherosclerosis. AB - Animal models of vein graft disease are used as preliminary tools to study and understand the pathogenesis of the disease in humans and improve its diagnosis, prevention and therapy. Several animal models that manifest lesions resembling neointimal hyperplasia of human vein grafts have been developed, but there are limitations in studying the mechanism of this disease in these models. We previously established a mouse model of vein bypass graft atherosclerosis that allows us to take advantage of transgenic and knockout techniques. Using this model, we studied the pathogenesis of vein graft atherosclerosis. The lesion in the grafts was characterised by mononuclear cell infiltration followed by smooth muscle cell (SMC) proliferation and matrix protein deposition, which is similar to the human lesion. Studies of the molecular mechanism of pathogenesis in this model revealed that physical force initiated signal pathways, particularly mitogen-activated protein kinases (MAPK), leading to vascular cell death and an inflammatory response, followed by SMC proliferation, which contributed to the development of arteriosclerosis. Suramin inhibited SMC migration and proliferation in vivo and in vitro by blocking platelet-derived growth factor (PDGF)-initiated PDGF receptor activation and MAPK-AP-1 signalling, and was also effective in inhibition of neointima hyperplasia in mouse vein bypass grafts. This new mouse model of vein bypass graft atherosclerosis affords us with a valuable new approach to attain further understanding of the mechanism of vein graft disease with the use of transgenic mice, and in evaluating the effects of drugs and gene therapy on vascular diseases. PMID- 16352096 TI - Cardiac sarcoidosis: an elusive diagnosis. AB - The diagnosis of sarcoidosis of the heart can be elusive, and is seldom established in life. We report the case of a 43-year-old man who underwent heart transplantation for presumed idiopathic dilated cardiomyopathy. Endomyocardial biopsy before transplantation showed only a mild infiltrate of lymphocytes. Histology of his explanted heart revealed extensive noncaseating granulomas and scarring, typical of sarcoidosis. A diagnosis of sarcoidosis had been made several years before by mediastinoscopic biopsy, after routine chest X-ray revealed mediastinal lymphadenopathy. Aside from the cardiac manifestations, the patient had no other symptoms of this disease. We discuss the inherent difficulties in the diagnosis of this rare but important condition, its varying presentations relating to the underlying pathology, as well as treatment options, including the role of transplantation. PMID- 16352097 TI - Hearts and minds. PMID- 16352098 TI - Myocardial energetics: still only the tip of an iceberg. PMID- 16352099 TI - Why publish hypothesis papers? PMID- 16352100 TI - Ischaemic preconditioning: developing knowledge of a developing process. PMID- 16352101 TI - Every heart beat is under neural command: an hypothesis relating to the cardiac rhythm. AB - In the human heart there is a sequential contraction of the systemic veins, systemic venous sinus and the pectinated right atrium, 'the systemic waltz', and sequential contraction of the pulmonary veins, pulmonary venous sinus and pectinated left atrium, 'the pulmonary waltz'. The systemic veins contract earlier than the pulmonary veins creating a 'duet. We hypothesise that this waltz and duet point to a complex extracardiac control of the cardiac rhythm on a beat to-beat neural basis. PMID- 16352102 TI - Alternative view on the mechanism of cardiac rhythmogenesis. AB - BACKGROUND: This article advances an hypothesis that there is duplication of the heart rhythmogenesis system. METHODS AND RESULTS: The following article reviews available data and advances an hypothesis to suggest new ideas about the mechanisms of cardiac rhythm generation. The hypothesis is that along with the existence of an intracardiac pacemaker, the generator of cardiac rhythm exists in the central nervous system - in the efferent structures of the cardiac centre of the medulla oblongata. Originating in the medulla oblongata, neural signals in the form of brief bursts of pulses are conducted to the heart along the vagus nerves and, by interacting with the cardiac pacemaker structures, cause generation excitement in the heart in exact accordance with the frequency of the bursts. The intracardiac rhythm generator is a life-providing factor that maintains the heart pumping function when the central nervous system is in a stage of deep inhibition (e.g. under anaesthesia or when unconscious). The central generator is the factor that provides the heart with adaptive reactions in natural conditions of the organism. CONCLUSIONS: Multi-leveled organisation of the mechanisms responsible for rhythmogenesis guarantees reliability and functional perfection of the cardiac rhythm generation system. PMID- 16352103 TI - Imaging cardiac metabolism in heart failure: the potential of NMR spectroscopy in the era of metabolism revisited. AB - Nuclear magnetic resonance (NMR) spectroscopy remains an extremely powerful technique for investigating abnormalities in the failing heart. The nondestructive nature of the technique allows the response to physiological, pathophysiological and pharmacological interventions to be studied within the same heart. Phosphorus-31 NMR has provided a gold standard over the past two decades for assessing the myocardial energy status both in vitro and in vivo. Carbon-13 isotopomer analysis is emerging as a direct way to monitor metabolic pathways and, in particular, investigate adaptations in energy provision in pathophysiological conditions. Using models of cardiac hypertrophy and heart failure, we investigated the sequences of changes in substrate oxidation in relation to function using 13C methods. The changes in metabolism modify the balance between energy provision and utilisation, and thus play a deleterious role in the progression towards decompensated heart failure. The application of NMR spectroscopy (phosphorus-31 and carbon-13) to the study of integrated metabolism is an area of research which is now coming into its own. Together with other new technologies, NMR will contribute to our improved understanding of cardiac metabolism in situ, leading to more rapid advances in targeting new therapeutic end points. PMID- 16352104 TI - Evolving strategies for surgical management of patients with severe left ventricular dysfunction. AB - As a result of an increasing population with advanced congestive heart failure and the lack of growth in cardiac transplantation, surgical treatments for heart failure have been re-examined. These therapies represent the evolution of well known operations such as coronary bypass surgery and valve surgeries, and the more novel left ventricular reconstruction and operations aimed at inhibiting left ventricular remodeling. When performed by surgeons with experience in this evolving speciality within cardiovascular surgery, surgery for advanced heart failure is a treatment of choice for many patients. PMID- 16352105 TI - Absence of apparent cardioprotection following ischemic preconditioning in immature rabbit hearts. AB - BACKGROUND: The cardioprotective effects of ischemic preconditioning (IPC) have been demonstrated in adult hearts of all species so far studied. AIMS: To investigate whether IPC could protect immature rabbit hearts against ischemia reperfusion injury in an isolated Langendorff-mode perfusion model. METHODS: Hearts from 18 rabbits aged 14-21 days were randomly divided into two groups (an IPC group and a control group). After isolated Langendorff-mode perfused hearts were equilibrated the IPC stimulus in the IPC group was 5 min global ischemia followed by 10 min reperfusion. Hearts in both groups were then made globally ischemic for 30 min (no perfusion), followed by 40 min reperfusion. Coronary flow (CF), heart rate (HR), left ventricular developed pressure (LVDP), and +/ dp/dt(max) were monitored at equilibration (baseline value) and then 5, 10, 20, 30 and 40 min after reperfusion. The values obtained after reperfusion were expressed as a percentage of their baseline value. Arrhythmia severity, myocardial enzymes in the coronary effluent, and myocardial energy metabolism were also determined. RESULTS: There were no statistical differences in postreperfusion CF, HR, LVDP and +/-dp/dt(max) between the two groups. Arrhythmia scores were also comparable between the two groups. The myocardial-specific isoenzyme of creatine kinase (CK-MB) leakage in the IPC group was increased, but not significantly different from that in the control group. At the end of reperfusion, the adenosine triphosphate (ATP) level in the myocardium in the IPC group was significantly lower than the level in the control group. CONCLUSIONS: This model of IPC was not able to protect juvenile rabbit hearts from ischemia reperfusion injury. This study suggests that signal transduction pathways involved in triggering cardioprotective mechanisms may not be fully developed, or are not able to be invoked by the present model in juvenile rabbit. PMID- 16352106 TI - The streptozotocin-diabetic rat as a model of the chronic complications of human diabetes. AB - BACKGROUND: Diabetes in humans induces chronic complications such as cardiovascular damage, cataracts and retinopathy, nephropathy and polyneuropathy. The most common animal model of human diabetes is streptozotocin (STZ)-induced diabetes in the rat. METHODS: This project assessed cardiovascular, ocular and neuropathic changes over a period of 24 weeks post STZ administration in rats. RESULTS: STZ-diabetic rats (n = 96) showed stable signs of diabetes (hyperglycaemia, increased water and food intake with no increase in bodyweight): 52% of untreated STZ-diabetic rats (n = 50) survived 24 weeks after STZ administration. STZ-diabetic rats were normotensive with slowly developing systolic and diastolic dysfunction and an increased ventricular stiffness. Ventricular action potential durations were markedly prolonged. STZ-diabetic rats developed stable tactile allodynia. Cataracts developed to presumed blindness at 16 weeks but proliferative retinopathy was not observed even after 24 weeks. CONCLUSION: The chronic STZ-diabetic rat mimics many but not all of the chronic complications observed in the diabetic human. The chronic STZ-diabetic rat may be a useful model to test therapeutic approaches for amelioration of chronic diabetic complications in humans. PMID- 16352107 TI - Assessing experimental models in myocardial injury: lack of activation of the proteases TACE and calpain in brief ischaemia and reperfusion. AB - BACKGROUND: Calpain inhibitors are reportedly cardioprotective. Furthermore, oxidative stress may acutely activate the sheddase tumour necrosis factor (TNF) alpha-cleaving enzyme (TACE). The aim of this study was to examine whether myocardial reperfusion leads to activation of the proteases mu- and m-calpain, and to evaluate which cardiac cells act as a source of TNF-alpha. METHODS: Isolated hearts (guinea pig) were subjected to global ischaemia (15 min) and reperfused. Calpain activity was determined by zymography. Calpastatin (inhibitor) and troponin I (substrate) were quantified by western blotting. Immunohistology of hearts and a human mast cell line (HMC-1) was used to localise expression of TNF-alpha and TACE. Shedding of TNF-alpha was assessed in Mono Mach, Jurkat-T, HMC-1 and peripheral blood leucocytes with and without oxidative stress. RESULTS: Neither of the ubiquitous calpains (mu- and m-calpain) was significantly activated by brief ischaemia/reperfusion, nor were calpastatin and troponin degraded more than in extracts of control hearts. Cardiac TNF-alpha immunoreactivity was localised to mast cells. None of the tested cell lines shed TNF-alpha in response to non-toxic amounts of oxidants. However, HMC-1 cells showed poor expression of proTNF-alpha, while TACE was abundant. CONCLUSIONS: Although the severity of ischaemia in the current model may have been insufficient, activation of calpain by ischaemia/reperfusion cannot be demonstrated simply in the Langendorff-mode perfused isolated heart. Mast cells are the prime source of myocardial TNF-alpha. A suitable whole-cell model remains to be found to demonstrate acute oxidative activation of TACE. PMID- 16352108 TI - Myocardial heat shock protein changes in the failing heart following coronary artery ligation. AB - BACKGROUND: Production of several heat shock proteins (Hsp) is enhanced after exposure to stress. There is little information concerning changes in myocardial Hsp under pathophysiological conditions. The aim of this study was to determine alterations in Hsp content in the viable left ventricular myocardium during the development of heart failure following coronary artery ligation (CAL). METHODS: Myocardial infarction was produced by CAL of Wistar rats. One and eight weeks after the operation, haemodynamic parameters of rats with CAL were determined and then expression of Hsp27, Hsp60 and Hsp72 was measured by western blotting. RESULTS: Animals showed a decrease in cardiac output and an increase in left ventricular end-diastolic pressure, symptoms of chronic heart failure (CHF), 8 weeks after CAL. Myocardial Hsp27 and Hsp72 at 1 week after CAL significantly increased, whereas expression of both proteins at 8 weeks was similar to that in rats which underwent a sham operation (without coronary artery ligation). In contrast, Hsp60 at 8 weeks, but not at 1 week, significantly increased in the sham rats. CONCLUSIONS: Diverse changes in myocardial Hsp occurred during the development of CHF. PMID- 16352109 TI - Coronary artery stent thrombosis associated with exercise testing. AB - Chest pain following coronary artery stenting is common, yet finding the cause can be difficult. Exercise testing has long been used in the assessment of chest pain, but its usefulness in patients who have recently undergone coronary artery stenting is in doubt. A case of exercise testing appearing to precipitate acute stent thrombosis in a patient several weeks post-coronary artery stenting is reported and compared to a similar case in the literature. The role of exercise testing in the assessment of chest pain early after coronary artery stenting is then reviewed. PMID- 16352110 TI - Anomalous origin of the left anterior descending artery from the pulmonary artery associated with bicuspid aortic valve and aortic coarctation. AB - A 15-year-old boy presented with exertional palpitations and chest pain. Investigation revealed anomalous origin of his left anterior descending coronary artery from his pulmonary trunk causing myocardial ischaemia. He previously had aortic coarctation repair with known aortic root dilation and a bicuspid aortic valve. His left anterior descending artery was implanted into the aortic root using a Gortex interposition conduit. This represents an interesting combination of cardiac abnormalities for which repair required consideration of the requirement of further surgery in the future. PMID- 16352111 TI - Thrombolysis for valve prosthesis. PMID- 16352112 TI - The cardiothoracic surgery workforce. PMID- 16352113 TI - Metabolic syndrome: the major consequence of obesity. PMID- 16352114 TI - Multiple mechanisms involved in obesity-induced hypertension. AB - Obesity and hypertension are two major cardiovascular risk factors gaining epidemic proportions in our modern world. The interplay between hypertension, obesity and their major detrimental outcome, cardiovascular disease, is multifaceted and can be represented as the three corners of a triangle. Obesity and hypertension both lead to cardiovascular complications. In addition, obesity per se can promote hypertension. In turn, cardiovascular diseases can also predispose to obesity and hypertension. Low levels of physical activity due to a weakened heart promote weight gain. Endothelial, vascular and renal dysfunctions, all consequences of high blood pressure, further worsen hypertension. The loop of mutually amplifying detrimental effects is thus closed: a 'vicious triangle' is established. The association between obesity and hypertension was recognised and described almost a century ago, but the mechanisms that underlie this connection are still not fully understood. Vasoconstriction and sodium retention seem to be the cornerstones of the obesity-hypertension puzzle. However, pathways possibly leading to vasoconstriction and sodium retention are numerous. Evidence has been gathered that hyperleptinaemia, hyperinsulinaemia and elevated free fatty acids may induce sympathetic activation and vasoconstriction. The latter is further potentiated by insulin resistance and endothelial dysfunction. Positive sodium balance and ensuing volume expansion may be due to increased renal tubular sodium reabsorption induced by sympathetic stimulation, insulin or by a hyperactive renin-angiotensin system. All enumerated factors act together toward a state of permanently elevated blood pressure. PMID- 16352115 TI - CHART: congestive cardiac failure in hospitals, an Australian review of treatment. AB - Despite strong evidence supporting the use of angiotensin-converting enzyme inhibitors (ACED, beta-blockers, and spironolactone in heart failure, evidence suggests these drugs are under-used and under-dosed. The aim of the present study was to determine the impact of hospitalisation on heart failure pharmacotherapy in patients with congestive heart failure (CHF). A retrospective study was conducted, based on 300 consecutive admissions with the medical record diagnosis of heart failure, in each of seven grade one teaching hospitals. At admission, 49.5% of patients were treated with ACEI, 19.2% with beta-blockers and 8.1% with spironolactone. Twenty-six per cent of untreated patients started ACEI treatment during their hospital stay, and 9.4% started beta-blockers The main determinants of treatment with ACEI at discharge were a primary diagnosis of heart failure (odds ratio (OR) = 1.886) and the presence of a potential contraindication (high creatinine OR = 0.458, cough OR = 0.187, renal artery stenosis OR = 0.309). Patients were less likely to be discharged on beta-blockers if greater than 85 years of age (OR = 0.545), or there was mention of airways disease (OR = 0.347), asthma (OR = 0.238) or type 2 diabetes (OR = 0.721) on the medical record. Patients admitted by a cardiologist were more likely to be discharged on beta blockers (OR = 3.207). Spironolactone was more likely used in patients with primary diagnosis of heart failure (OR = 1.549), aged less than 85 years (OR = 0.319), and/or admitted by a cardiologist (OR = 1.827). The substantial number of patients admitted to hospital with a secondary diagnosis of heart failure should be targeted for therapeutic optimisation. PMID- 16352116 TI - Extended follow up after isolated aortic valve replacement in the elderly. AB - The present paper reviews the extended follow up of all patients aged >/=70 who underwent isolated aortic valve replacement at our institution in the 1980s. Patients were identified from the surgical database and clinical information was gathered. Long-term follow up information was obtained from the patient, their family, or doctor. Ninety-three patients aged >/=70 years (median 73, range 70 80) comprised the study population. The indication for surgery was aortic stenosis in 68 patients (73%). Fifty-two patients (56%) received an allograft valve, 17 (18%) a bioprosthetic valve, and 24 (26%) a mechanical prosthesis. The median hospital stay was 12 days (range 0-105 days). Five surgical deaths occurred. Detailed follow up was obtained for the 71 patients who died later, and the 16 living patients (one patient lost). The median length of follow up was 6.8 years (range 0.1-18.9 years). Patients who received an allograft aortic valve had a significantly better long-term survival (median 10.6, 95% confidence intervals (CI) 8.1-13.8 years) compared to those receiving mechanical or bioprosthetic valves (median 6.5, 95% CI 4.7-11.9 years), P = 0.03. For the entire group, survival was similar to the age- and sex-matched population. Of the 16 patients alive at follow up (mean age 87, range 83-92), most were free of angina (12, 75%) and heart failure (10, 63%). The conclusion from the current study is that isolated aortic valve surgery in the elderly, particularly with an allograft valve, is associated with an excellent long-term outcome. A survival benefit was demonstrated comparing allograft aortic valve replacement to other valve types. PMID- 16352117 TI - Atrial myxoma with coexistent coronary artery disease: a report of two cases. AB - The age at presentation and the symptoms of atrial myxomas and coronary artery disease can be similar. At times, the two lesions coexist. Operative strategy needs to be carefully planned when combined surgical treatment is contemplated. In the present paper, two cases of concomitant atrial myxoma and atherosclerotic coronary artery disease are reported. The first patient had a left atrial myxoma with triple-vessel coronary artery disease. The other had a right atrial myxoma and needed two bypass grafts. Both patients had a satisfactory outcome. PMID- 16352118 TI - Cardiac sarcoidosis: an elusive diagnosis. PMID- 16352119 TI - Diastolic heart failure: can we afford to be in diastole? PMID- 16352120 TI - Off-pump coronary artery bypass: where are we? PMID- 16352121 TI - Anticoagulation and mechanical heart valves: extrapolation has its hazards. PMID- 16352122 TI - Is 'diastolic heart failure' a diagnosis of exclusion? Echocardiographic parameters of diastolic dysfunction in patients with heart failure and normal systolic function. AB - BACKGROUND: Diastolic heart failure (DHF) is reported to account for 30-50% of heart failure presentations, but its prevalence in the absence of overt coronary disease is unclear. Diastolic heart failure is usually defined by exclusion (heart failure with normal left ventricular (LV) systolic function), and few studies have sought a specific diagnosis of diastolic dysfunction. The objective of the present study was to determine the prevalence of isolated DHF and characterise LV diastolic function in patients without clinical evidence of coronary disease, who were referred for LV function assessment. METHODS: Among 938 consecutive patients referred for assessment of LV function, diastolic dysfunction was sought in patients with clinical heart failure, normal systolic function, and no valvular or coronary disease. The evaluation was based on measurement of early (E) and late (A) transmitral velocities and E wave deceleration time (DT). Pulmonary vein systolic, diastolic and atrial reversal velocities were used to differentiate pseudonormal filling in patients with normal E/A and DT. RESULTS: Normal LV systolic function was present in 331 patients (35%), of whom 53 (6%) met criteria for a clinical diagnosis of DHF. Diastolic dysfunction was confirmed by echocardiography in 38 patients (72% of clinical DHF patients), of whom 27 had impaired LV relaxation, 10 had pseudonormal filling, and one had restrictive filling. Diastolic function was normal in 13 and indeterminate in two patients. Pseudonormal or restrictive LV filling were more prevalent in patients with acute heart failure (7/20, P < 0.05). CONCLUSIONS: Carefully defined, isolated DHF is uncommon, but most of these patients demonstrate echocardiographic evidence of diastolic dysfunction. PMID- 16352123 TI - Nitric oxide does not mediate the vasodilation of early human pregnancy. AB - BACKGROUND: In early pregnancy, a substantial drop in arterial blood pressure occurs, that might be attributed to enhanced vascular nitric oxide synthesis. We investigated whether nitric oxide mediates the vasodilation that occurs in early human pregnancy. METHODS: Resting and stimulated forearm vascular resistance were measured (venous occlusion plethysmograph) in six women at 10 +/- 3 weeks of uncomplicated pregnancy and in the same women 7 +/- 5 weeks after elective termination of pregnancy. Forearm vascular resistance was also measured in six non-pregnant, healthy controls. RESULTS: Resting forearm vascular resistance was similar during pregnancy (33 +/- 16 arbitrary units (AU)), after pregnancy (31 +/ 10 AU) and in controls (41 +/- 13 AU, P > 0.05). The decreases in forearm vascular resistance to intrabrachial infusions of acetylcholine (2 and 20 microg/min), serotonin (10 and 100 ng/min) and sodium nitroprusside (1 and 2.5 microg/min) were similar in all groups. The nitric oxide synthase inhibitor NG monomethyl-L-arginine (16 micromol/min) produced similar increases in vascular resistance in pregnant women (38 +/- 17 AU), after pregnancy (36 +/- 14 AU) and in control subjects (42 +/- 8 AU, P = NS). CONCLUSIONS: These results indicate that neither basal nor stimulated nitric oxide levels are altered in the forearm circulation during first trimester pregnancy. PMID- 16352124 TI - Quantification of haemodynamic performance of stented and stentless aortic bioprostheses and potential influence on survival. AB - BACKGROUND: The goal of aortic valve replacement is relief of symptoms, optimisation of haemodynamics, regression of left ventricular mass and advancement of survival. The objective of this review article is to provide the evidence-to-date on the clinical performance of stented and stentless heterograft bioprostheses with regard to haemodynamics, durability and survival. METHODS: The haemodynamic advantage of aortic valve replacement prostheses is judged on ability to minimise postoperative gradients and to optimise the normalisation of left ventricular mass and function. The most frequent cause of high postoperative gradients occurs when the effective prosthetic valve area is less than that of the normal human valve. The effective orifice area index (EOAI) of >/= 0.85 cm(2)/m(2) is considered optimal to prevent patient-prosthesis mismatch (PPM) at rest and exercise. RESULTS: The stented bioprostheses contribute to obstructive non-physiological flow patterns whereas stentless bioprostheses provide laminar non-obstructive flow. The stentless bioprostheses have been shown to have larger effective orifice areas and lower gradients. Mismatch is decreased with stentless bioprostheses especially when prosthesis size is 0.8 cm(2)/m(2) provides the best long-term regression of LVM. It has been identified that a tendency for PPM in sizes 21 and 23 mm stented bioprostheses did not prevent adequate achievement of appropriate LVMI. Survival at 5 years favoured stentless over stented bioprostheses for patients <70 years, but not in patients >/= _70 years of age. The durability comparison of the stentless bioprostheses must wait until 10-15 years experience is achieved. There is preliminary evidence that uneven shear stress on the leaflet of a regurgitant stentless bioprosthesis can accelerate leaflet tears at the level of the commissures. Dilation of the aortic root and, particularly, the sinotubular junction, can cause progressive stentless valve insufficiency. CONCLUSIONS: The long-term performance advantages or disadvantages of stentless bioprostheses compared to stented bioprostheses will require at least another 5-7 years of cumulative stentless bioprostheses experience. Surgeons can use an algorithm intraoperatively to prevent patient-prosthesis mismatch while choosing the optimal prosthesis. PMID- 16352125 TI - Five year clinical follow up of patients who have had off pump coronary artery bypass grafting. AB - BACKGROUND: Off pump coronary artery bypass (OPCAB) has become a common technique for conducting coronary artery surgery. There has been some concern that the anastomoses might not be conducted as effectively on the beating heart, which could lead to poor long-term results. The aim of the present study was to follow up all patients who had undergone OPCAB at our institution to determine clinical outcomes up to 5 years postoperatively. METHODS: All living patients who had undergone OPCAB up until December 2000 were telephoned. For those who could be contacted, a detailed questionnaire was completed and the data were analysed. RESULTS: The technique of OPCAB was carried out on 312 patients. Thirteen of these patients died, five perioperatively. Actuarial survival at 5 years was 94.6% and freedom from cardiac related events was 92.1%. There were only two patients with angina worse than class I. Patients showed a high rate of compliance with risk factor management. Forty per cent of patients claimed to have had psychological problems related to the surgery. CONCLUSIONS: Mid-term results of OPCAB are very satisfactory, but randomised trials are needed to see whether they are different from the results of conventional coronary artery bypass grafting. PMID- 16352126 TI - Five year clinical follow up of patients who have off-pump coronary artery bypass grafting. PMID- 16352127 TI - Lower anticoagulation for mechanical heart valves: experience with the ATS bileaflet valve. AB - BACKGROUND: The design of the bileaflet ATS (ATS Medical Inc., Minneapolis, USA) mechanical valve incorporates an open pivot at the hinge mechanism. Total washout of the blood at the pivot area was observed using 3-D computational fluid dynamics modelling. This phenomenon could make the valve less vulnerable to clot formation in patients without major thromboembolic risk factors. METHODS: From January 1993 to June 1999, 286 consecutive patients had the ATS valve inserted in the aortic position. Patients were divided into two groups. Group 1 comprised all patients in regular sinus rhythm with good left ventricular function (144 patients). Group 2 included patients in non-sinus rhythm and/or with large hypocontractile left ventricles (142 patients). The anticoagulation regime in group 1 was used to obtain an international normalised ratio (INR) between 1.5 and 2.5. This contrasts with our regular aim to maintain the INR strictly between 2.5 and 3.5 for all mechanical valves, as achieved in group 2. RESULTS: The follow-up period (99% completeness) ranged from 18 to 84 months. Survival (Kaplan Meier) was 97 and 98% and 92 and 81% at 1 and 5 years in group 1 and group 2, respectively (P = 0.12). Log rank analysis failed to detect a statistical difference in thromboembolism or bleeding between both groups (P > 0.05). However, trends were in favour of group 1. Univariate analysis selected poor ventricular function and an 'erratic' INR value (P = 0.002) as risk factors for death. The sole independent risk factor for bleeding was the use of aspirin (P = 0.025). CONCLUSIONS: The excellent group 1 data and outcome encouraged us to continue our low intensive anticoagulation regime and perhaps should be regarded as a new concept for treatment of selected mechanical valve patients. PMID- 16352128 TI - From classical sternotomy to truly endoscopic mitral valve surgery: a step by step procedure. AB - BACKGROUND: There is an increasing tendency towards minimally invasive valve surgery and various surgical techniques have been proposed to realise this goal. The aim of the present study was to describe our current surgical technique and clinical experience with respect to an endoscopic technique that allows the surgeon to perform an operation through a series of small intercostal ports. METHODS: After a learning experience with thoracoscopic left internal mammary to left anterior descending coronary artery bypass surgery, we adopted the endocardiopulmonary bypass technique to perform mitral valve surgery. The technique requires exclusive use of video-assisted surgery and control by transoesophageal echocardiography (TEE). Surgery requires long instruments and extra-corporeal knot tying. Between February 1997 and November 2001, 259 patients were operated on. Mitral valve repair was performed in 190 of them. One patient had a redo procedure using this approach to correct a paravalvular leak, but all other procedures were primary interventions. RESULTS: In all patients, surgery was performed using a 2 inch working port and two additional half-inch trocar ports. Five patients required a conversion to median sternotomy: three because of inadequate size of the femoral vessels and two because of intraoperative aortic dissection. Hospital mortality included two patients, and seven patients required late reoperation (four of these were as a result of endocarditis). CONCLUSIONS: Endoscopic mitral valve surgery is demanding, but feasible. Once the appropriate skills are acquired, both patient and surgeon can enjoy the benefits of this exciting new technique. PMID- 16352129 TI - Endogenous cardiac opioids: enkephalins in adaptation and protection of the heart. AB - Opiates have been used for thousands of years in the form of opium for relief of pain or fever and to induce sleep. However, it was only in the 1970s that the endogenous ligands for the opiate receptors were identified and termed opioid peptides. Opioid peptides activate G protein-coupled receptors in the central and autonomic nervous system, with marked effects on the regulation of pain perception, body temperature, respiration, heart rate and blood pressure. Cardiovascular regulatory effects of endogenous opioids were initially considered to originate from neural centres in the central nervous system, facilitating a regulatory role in neuro-transmission, as demonstrated by the presynaptic co release from sympathetic neurones of norepinephrine with enkephalin or acetylcholine with enkephalin. However, opioid peptides of myocardial origin have also recently been shown to play a key role in local regulation of the heart. This brief review highlights the key features of the enkephalin opioids in the heart and the current understanding of their role in development, ageing, cardioprotection, hypertension, hypertrophy, and heart failure. PMID- 16352130 TI - Association of a double orifice mitral valve with a bicuspid aortic valve in an explanted heart with dilated cardiomyopathy. PMID- 16352131 TI - Salvage stenting for acute renal failure secondary to renal artery stenosis. AB - We present two cases of successful, emergency renal artery stenting in the setting of acute renal failure requiring dialysis secondary to renal artery stenosis. Early reopening of the stenotic renal artery led to the resolution of acute renal failure and obviated the need for further dialysis in both cases. PMID- 16352132 TI - AIDS and heart disease: is cardiac surgery justified? AB - A compromised immune system, limited survival and increased risk to the anaesthetic and surgical team of acquiring HIV infection have been the major concerns in offering cardiac surgery to patients with AIDS. The current report presents a patient with AIDS who underwent successful, uncomplicated coronary artery surgery. He remains free of ischaemic and any infective symptoms 12 months postoperatively. PMID- 16352133 TI - Sudden death prevented in hypertrophic cardiomyopathy. AB - The most common cause of sudden cardiac death in individuals aged less than 35 years, including competitive athletes, is the inherited disorder hypertrophic cardiomyopathy (HCM). Until recently, no therapeutic intervention had been identified to prevent sudden death in HCM. This case report highlights the role of the implantable cardioverter defibrillator (ICD) in preventing sudden death in patients with HCM. The report highlights the importance of risk stratification in patients with HCM in order to identify those individuals who would most likely benefit from ICD therapy. The ICD implantation is now the treatment of choice in preventing sudden death in selected (highest risk) populations with HCM. In the future, understanding the molecular basis of sudden death in HCM may identify potential new gene-based therapeutic strategies. PMID- 16352134 TI - Asymmetric myocardial thickening during acute myocarditis. AB - The case of a young man presenting with an acute myocarditis is described. Transthoracic echocardiography showed severe asymmetric thickening of the posterolateral left ventricle with associated hypokinesis. These abnormalities resolved following the patient's recovery from the acute illness. Asymmetric posterolateral myocardial thickening has not previously been reported in acute myocarditis. PMID- 16352135 TI - Conflict of interest. PMID- 16352136 TI - Introduction. PMID- 16352137 TI - The contributions of Paul Wood to clinical cardiology. PMID- 16352139 TI - The beginnings of cardiology and cardiac surgery in Australia and New Zealand. PMID- 16352138 TI - History of early cardiac surgery in Australia. PMID- 16352140 TI - The early history of cardiac surgery in New Zealand. PMID- 16352141 TI - Lessons from history for the younger cardiologist. PMID- 16352142 TI - Victorian Heart Centre proceedings 2002. PMID- 16352143 TI - Epworth hospital interventional cardiology report, 2002. AB - In March 2001, the Victorian Heart Centre at Epworth Hospital implemented a database management system to improve the standard of data collection and analysis of procedures performed in our catheter laboratory. A retrospective analysis of procedures undertaken during 2000, together with prospective data collected during 2001/2002, has enabled us to achieve our goals of benchmarking performance and undertaking research on the outcomes of interventional cardiology procedures at Epworth. The figures reported in this year's cardiac catheterisation laboratory audit report confirm a continued high standard performance in our laboratory. PMID- 16352144 TI - Cardiac surgical audit Epworth Hospital 2002. AB - The cardiac surgical programme at Epworth Hospital has performed in excess of 13 000 open-heart surgical procedures between 1981 and 2003. During the audit period (2002) 411 patients underwent open-heart surgery. The 2002 Cardiac Surgical Audit confirms the very high standard of surgery. The goals for the future are to maintain the low complication rate and maintain an efficiency to reduce the financial burden on the hospital, the insurance companies and the health system. PMID- 16352145 TI - Noradrenaline: friend or foe? AB - Septic shock, systemic inflammation and pharmacological vasodilatation are often complicated by systemic hypotension despite aggressive fluid resuscitation and an increased cardiac output. If the physician wishes to restore arterial pressure to higher levels (> 80-85 mmHg), with the aim of sustaining cerebral and coronary perfusion pressure, the administration of systemic vasopressor agents, such as norepinephrine (noradrenaline), becomes necessary. However, because norepinephrine (NE) induces vasoconstriction in many vascular beds (visibly in the skin), it may decrease renal and visceral blood flow, impairing visceral organ function. This unproven fear deters clinicians from using NE more consistently. Vasodilated states, however, are often associated with impaired peripheral vascular responsiveness. In such states, unlike under normal circulatory conditions, NE may actually improve visceral organ blood flow by selectively increasing organ perfusion pressure. Data available from animal studies show that the increased organ perfusion pressures achieved with NE results in improved GFR and renal blood flow. In fact, recent sophisticated physiological analysis of its effects on the kidney shows that, even after controlling for the pressure effect, NE therapy is associated with an increase in renal blood flow after endotoxin administration. In particular, the renal Pzf (pressure at which there is no further blood flow) is decreased such that, at a constant pressure, renal blood flow increases after NE. There are no controlled human data to define the effects of NE on the kidney in the clinical context. However, many patient series have now been reported. They show a seemingly positive effect of NE administration on GFR and urine output. Our clinical experience in septic patients and cardiac patients with inflammatory or pharmacological vasodilatation is also positive. We have demonstrated a positive effect on coronary blood flow. There is no reason to fear the effect of NE. If it is used to support a vasodilated circulation after adequate intravascular filling has occurred and after a normal or increased cardiac output has been established, it is likely to be a friend not a foe. PMID- 16352146 TI - Developments in vascular surgery at Epworth Hospital. AB - Epworth Hospital has been a leading centre for vascular surgery in Melbourne for over 30 years with a strong focus on vascular reconstruction by open surgical methods. Epworth pioneered the hybrid suite, which combined state-of-the-art imaging, and operating theatre standard suite. This development has become the benchmark for other vascular centres when designing their own vascular suites. Epworth Hospital has become a leading centre in the southern hemisphere for endografting for both abdominal aortic aneurysm and thoracic aneurysm with 76 endografts being performed in the calendar year 2000, 125 in the year 2001 and 106 in the year 2002. We look forward to Epworth Vascular Unit continuing to practice world-best standards in peripheral arterial reconstruction, to continue its role in mentoring surgical trainees and other vascular surgeons learning endovascular skills and to promote the development of innovations in vascular surgery. PMID- 16352147 TI - Why do aneurysms burst? AB - Aneurysms are defined as bulges twice the diameter of the nearby normal vessel. Aneurysms burst when the artery wall becomes too weak for the blood pressure within. Urgent clinical examination by a vascular surgeon, then an ultrasound or CT angiogram if time permits before repair, may save life. The result of a timely repair at Royal Melbourne Hospital of Epworth is 90-95% success, while only 50% of patients survive an attempt to repair the aneurysm once it has burst. PMID- 16352148 TI - The principles of metabolic therapy for heart disease. AB - Metabolic therapy involves the administration of a substance normally found in the body to enhance a metabolic reaction within the cell. This may be achieved in two ways. First, for some systems, a substance can be given to achieve greater than normal levels in the body so as to drive an enzymic reaction in a preferred direction. Second, metabolic therapy may be used to correct an absolute or relative deficiency of a cellular component. Thus, metabolic therapy differs greatly from most standard cardiovascular pharmacologic therapy such as the use of ACE Inhibitors b-blockers, statins and calcium channel antagonists that are given to block rather than enhance cellular processes. In this review we highlight some metabolic substances that have potential benefit in treating heart disease or improving outcomes after cardiovascular interventions. Glucose-insulin potassium therapy is protective against myocardial ischaemia by elevating myocardial glycogen levels. Coenzyme Q(10) is a lipid-soluble antioxidant that plays a crucial role in cellular ATP production. Magnesium orotate, a key intermediate in the biosynthetic pathway of glycogen, has been shown to improve the energy status of the cell and improve recovery from cardioplegic arrest. The amino acid aspartate plays an important role in providing energy substrates for oxidative phosphorylation in the myocyte. By improving cellular energy production, metabolic therapy has the potential to benefit cardiac function during the stress of cardiac surgery, myocardial infarction and cardiac failure. PMID- 16352149 TI - Catheter closure of atrial septal and ventricular septal defects using the Amplatzer devices. AB - The Amplatzer Septal Occluder (ASO) is a device that combines the advantages of being a double-disc with a self-centering mechanism. It is the first and only device to ever receive full approval for clinical use in children and adults with secundum atrial septal defects (AD) from the United States Food and Drug Administration. It has been used successfully to close secundum ASDs, patent foramen avale and Fontan fenestrations. The first patient to undergo closure with the ASO was approximately 6 years ago. So far the mid-term results are very encouraging with no long-term complications for the presence of the device. Complications encountered with the use of the Amplatzer septal occluder are rare and most may be managed in the catheter laboratory. Most of the complications occur in the immediate period post-implantation. The Amplatzer muscular and membranous ventricular septal defect devices are still undergoing clinical trials in the United States; however, they are fully approved in many countries. Long term results about safety and efficacy, as well as results involving larger defects are being collected. PMID- 16352150 TI - Irish contributions to our understanding of heart disease. AB - Ireland has made an amazing contribution to our understanding of heart disease. From Robert Graves in 1796 to Jeremy Swan and Denis Burkitt in the 1990s the names of many Irishmen have become familiar to all of us. PMID- 16352151 TI - Atrial fibrillation. AB - Atrial fibrillation (AF) is a hetrogeneous disease that may present in many forms. Advances in our understanding of the mechanism responsible for AF have led to exciting new therapies that may soon provide a definitive cure. PMID- 16352152 TI - Introducing the integrated cardiovascular services at Dr Hasan Sadikin Hospital. AB - Cardiovascular disease is a growing problem in Indonesia. We are still in our infancy and there are still numerous problems that need to be overcome. The main issues at this stage are finding ways for cardiac procedures to be affordable for patients and to improve the capabilities and knowledge of our staff. PMID- 16352153 TI - Drug-induced prolonged repolarisation (acquired long QT syndrome) arrhythmias. AB - Long QT Syndrome is a genetically determined disease of protein structures in the heart called ionic channels that control the flow of ions such as sodium, potassium and calcium, which produce the electrical activity of heart, depolarisation and repolarisation. Measurement of the QT interval on the electrocardiogram (ECG) can be quite difficult. Treatment of TdP due to ALQTS entails withdrawal of any precipitating agent, administration of MgSO(4) and potassium supplements. Correction of bradycardia is very important and temporary pacing may be necessary. If an antiarrhythmic agent is required after all the above, then lignocaine can be used. PMID- 16352154 TI - The twilight zone of troponins. AB - Since its introduction into the clinical environment in the early nineties, the cardiac enzyme troponin has significantly changed the way we diagnose and manage acute coronary syndromes. Troponin I is a biochemical marker of myocardial injury with a high level of specificity and sensitivity. It has been demonstrated that as ischaemia progresses, troponin I is degraded predictably into smaller and smaller fragments that can be detected in the blood-stream. This may eventually allow more accurate determination of the duration of ischaemia and the likelihood of myocardial salvage and recovery. PMID- 16352155 TI - Diabesity in Australia: an affair of the heart. AB - The objective of the Australian Diabetes and Lifestyle Study (AusDiab) was to determine the frequency of diabetes and other categories of glucose intolerance (impaired glucose tolerance (IGT) and impaired fasting glucose (IFG)), as well as other cardiovascular risk factors in Australia and to compare the prevalence with previous Australia data. The study involved a national sample involving 11 247 participants aged 25 years from the six states and the Northern Territory. They were examined in a cross-sectional survey using the 75-g oral glucose tolerance test to assess fasting and 2-h plasma glucose levels. The World Health Organization diagnostic criteria were used to determine the prevalence of abnormal glucose tolerance. The prevalence of diabetes in Australia was 8.0% in men and 6.8% in women, and an additional 17.4% of men and 15.4% of women had IGT or IFG. The overall diabetes prevalence in Australia was 7.4%, and an additional 16.4% had IGT or IFG. Diabetes prevalence has more than doubled since 1981, and this is only partially explained by changes in age profile and obesity. Almost one in four Australians 25 years and over has either diabetes or a condition of impaired glucose metabolism. This condition is associated with substantially increased immediate risk of heart disease as well as increased risk of diabetes in the future. In addition, there were high prevalences of other key cardiovascular disease risk factors. Australia has a rapidly rising prevalence of diabetes and other categories of abnormal glucose tolerance. The prevalence of abnormal glucose tolerance in Australia is one of the highest yet reported from a developed nation with a predominantly European background. PMID- 16352157 TI - Changing leadership for the journal. PMID- 16352156 TI - Heart, lung and circulation: flagship or millstone? PMID- 16352158 TI - Heart, lung and circulation enters a new era. PMID- 16352160 TI - Surely polly'll behave and take her pills? The challenge of implementing guidelines. PMID- 16352161 TI - Radionuclide imaging for the detection of cardiac infection. PMID- 16352162 TI - The medical indemnity crisis, medical negligence and the urgent need for reform. PMID- 16352163 TI - Hypertrophic cardiomyopathy: from "heart tumour" to a complex molecular genetic disorder. AB - Hypertrophic cardiomyopathy (HCM) is a disorder which has fascinated clinicians for many years. The remarkable diversity in clinical presentations, ranging from no symptoms to severe heart failure and sudden cardiac death, illustrates the complexity of this disorder. Over the last decade, major advances have been made in our understanding of the molecular basis of several cardiac conditions. HCM was the first cardiac disorder in which a genetic basis was identified and as such, has acted as a paradigm for the study of an inherited cardiac disorder. At least eleven genes have now been identified, defects in which cause HCM. Most of these genes encode proteins which comprise the basic contractile unit of the heart, i.e. the sarcomere. Genetic studies are now beginning to have a major impact on diagnosis in HCM, as well as in guiding treatment and preventative strategies. While much is known about which genes cause disease, relatively little is known about the molecular steps leading from the gene defect to the clinical phenotype, and what factors modify the expression of the mutant genes. Concurrent studies in cell culture and animal models of HCM are now beginning to shed light on the signalling pathways involved in HCM, and the role of both environmental and genetic modifying factors. Understanding these basic molecular mechanisms will ultimately improve our knowledge of the basic biology of heart muscle function, and will therefore provide new avenues for diagnosis and treatment not only for HCM, but for a range of cardiovascular diseases in man. PMID- 16352164 TI - Post-infarct left ventricular free wall rupture-not always a lethal complication of acute myocardial infarction. AB - Post-myocardial infarction cardiac rupture is an important complication and cause of death in the period following myocardial infarction. It is rarely diagnosed before death. However, early diagnosis is crucial as successful treatment is possible with surgery. A successful outcome is sometimes compromised by difficult anatomy or an extensive infarct. Presentation, diagnosis and treatment of cardiac rupture is reviewed in this article, and is illustrated by five cases of cardiac rupture. PMID- 16352165 TI - Early definition of treatment outcomes after reperfusion therapy for myocardial infarction. AB - AIMS: Early definition of treatment outcomes, including coronary patency and infarct size, after reperfusion therapy for myocardial infarction (MI) is desirable to identify patients requiring further intervention. METHODS AND RESULTS: Patients receiving reperfusion therapy for a first MI had continuous 12 lead ST segment monitoring to document reperfusion and ischaemia time. Infarct size was measured by 12-lead QRS score and radionuclide scintigraphy ((201)Tl single-photon emission computed tomography, SPECT) at 1 week, and left ventricular function by echocardiography at 1 week and 1 month. Resolution of ST elevation accurately detected TIMI 2 or 3 reperfusion (predictive accuracy 93%) in 55 patients undergoing immediate angioplasty, but ST recovery was delayed (17+/-14min) after angiographic reperfusion. A multivariate model, including risk region and ischaemia time, accurately predicted MI size (R(2)=0.80, P<0.00001) in these patients. The same model, prospectively applied on Day 1 to 154 patients receiving thrombolytic therapy, accurately predicted MI size, measured by QRS score (R(2)=0.88, P<0.0000001) and (201)Tl SPECT (R(2)=0.75, P<0.000001) at 1 week for individual patients. Regional myocardial wall motion at 1 month was directly correlated with MI size predicted by the model on Day 1 (r=0.73, P<0.0001). CONCLUSIONS: Use of ST segment monitoring during reperfusion therapy facilitates early prediction of treatment outcomes, including coronary reperfusion, infarct size and ventricular function. PMID- 16352166 TI - Diltiazem reverses tissue Doppler velocity abnormalities in pre-clinical hypertrophic cardiomyopathy. AB - BACKGROUND: Abnormalities in systolic and diastolic function shown by tissue Doppler imaging have been shown to be present in patients with hypertrophic cardiomyopathy who do not yet show clinical or echocardiographic evidence of the disease. These become more marked as left ventricular hypertrophy develops. We attempted to show that these abnormalities could be reversed by treatment with diltiazem. METHODS AND RESULTS: Six adults, who were carriers of a mutation involving the cardiac myosin binding, protein-C gene and who did not show clinical electrocardiographic or echocardiographic evidence of the disease were given a dose of 240mg of diltiazem daily. Tissue Doppler peak systolic and early diastolic velocities at the lateral mitral annulus were examined before treatment and at a mean of 8 weeks after starting treatment. Improvement in both parameters occurred with early diastolic velocities returning to normal and most systolic velocities also becoming normal. CONCLUSION: Diltiazem may have a role in helping to prevent abnormalities of function and perhaps the development of left ventricular hypertrophy in patients with pre-clinical hypertrophic cardiomyopathy. PMID- 16352167 TI - Allograft aortic valve replacement in the adult: a review. AB - Aortic valve replacement using an allograft has been used continuously for over 40 years. Its advantages are excellent haemodynamic function, low thrombogenicity, resistance to infection and avoidance of the complications of anticoagulation. The main concern is its long-term durability, with the high hazard phase for failure between 10 and 20 years. We have only recently been able to judge the true long-term behaviour of the contemporary allograft with two recently published series of patients having reached follow-up beyond 20 years in significant numbers. This review of allograft aortic valve replacement in the adult covers the areas of history, benefits, techniques of sterilisation and preservation, operative methods and outcomes. PMID- 16352168 TI - Alternative for primary pericardial closure: sentry for re-entry. AB - BACKGROUND: Redo cardiac surgery is considered high-risk surgery as accidental injury to the aorta, the innominate vein, the ventricles and the atria is a possibility. Such accidental injury occurs when the cardiac chamber is adherent to the undersurface of the sternum. Closure of pericardium at the time of primary surgery can prevent adherence of cardiac chambers to the sternum, but may increase the risk of tamponade. This study aimed to show that covering heart with a pedicled pericardial fat pad not only serves the purpose of cover but also avoids the adverse haemodynamic effects of primary pericardial closure. METHODS: Forty patients undergoing elective cardiac surgery were randomised into two groups depending on the way pericardium was managed. Both techniques were already in routine use in our unit and in other units around the country. One method is to leave the pericardium widely open, the other is to loosely oppose the pericardial fat pad over the surface of the aorta and right ventricle. Twenty three patients had a pedicled pericardial fat pad covering the heart: Closure Group. Seventeen patients had no pericardial fat pad cover over the heart: Open Group. A haemostasis clip was used as a radio-opaque marker over the epicardium in both groups. Post-operation heart rate, central venous pressure, pulmonary artery diastolic pressure, mean arterial pressure and cardiac index were measured and recorded 1, 3 and 8h after surgery. The distance between the haemoclip and the posterior table of the sternum was measured at 6 days and 6 months post operation. Haemodynamic parameters and the retrosternal space depth were compared between the two groups. RESULTS: There were no important differences in haemodynamic parameters between the two groups. Post-operative lateral chest Roentgenograms showed that the distance between epicardial surface and the posterior table of sternum was larger in the Closure Group compared to Open Group on post-operative day 6, 17.5+/-1.0mm versus 13.4+/-1.3mm (P=0.0013) and 6 months later, 12.3+/-0.8mm versus 6.0+/-1.2mm (P<0.001). There was no mortality in either group. CONCLUSION: Pedicled pericardial fat pad cover is a good alternative to primary pericardial closure as there are no adverse haemodynamic effects in early post-operative period and the long-term benefit of protection of heart at the time of re-sternotomy can be expected. PMID- 16352169 TI - Viable extracardiac total right heart bypass: technical considerations and mid term results. AB - BACKGROUND: Direct connection of both caval veins to the pulmonary arterial system would be the ideal right heart bypass by virtue of elimination of prosthetic material and extensive atrial suture lines. However, it can be applied only to selected patients with peculiar anatomical arrangements. We decided to perform total extracardiac right heart bypass using pedicled in situ pericardium in situations when a direct connection was not possible. METHODS: A cohort of five patients, aged 3.5, 4, 10, 10 and 31 years, respectively, with complex cyanotic congenital heart diseases were candidated to extracardiac total cavopulmonary connection using viable in situ pericardium. Aortic cross-clamp was completely avoided in all patients. RESULTS: There was no early or late death. Postoperatively, all patients are in functional class I and in normal sinus rhythm at a mean follow-up of 30 months (S.D.=+/-19 months). Postoperative serial echocardiograms revealed wide, unrestricted anastomoses, unobstructed venous flow to both pulmonary arteries with cardiac pulsations and respiratory variations in the pulmonary arteries due to compressible lateral tunnel. Cavopulmonary angiograms (n=5) in late postoperative period have demonstrated a smooth, nonturbulent, somewhat pulsatile flow without pressure gradient. CONCLUSIONS: The technical advantages and haemodynamic benefits of this operation are encouraging. A longer follow-up is needed to confirm the early results, especially late atrial dysrrhythmias, pulmonary venous obstruction, thromboembolic complications and growth of the conduit. PMID- 16352171 TI - Surgical treatment of late tricuspid regurgitation after left cardiac valve replacement. AB - BACKGROUND: The development of late tricuspid regurgitation (TR) following left cardiac valve replacement is an important complication, as it is associated with a severe impairment of exercise capacity and a poor symptomatic outcome. The pathogenesis of this condition remains poorly defined. It is still a challenge in terms of its prevention, treatment and indications for surgical correction. AIMS: To investigate the possible pathogenesis and report the surgical results of the late TR after left cardiac valve replacement. METHODS: There were 56 patients with moderate to severe TR after left cardiac valve replacement, divided into normal prosthesis group (10 patients with normal prosthetic valve function) and dysfunctional prosthesis group (46 patients with prosthetic valve dysfunction). In the normal prosthesis group, 4 patients underwent mitral valve replacement (MVR) and 6 patients underwent combined mitral and aortic valve replacement (DVR). Patients in the dysfunctional prosthesis group included MVR in 36, aortic valve replacement (AVR) in 4 and DVR in 6, with bioprosthetic valve dysfunction occurring in 18, mechanical prosthetic valve obstruction in 22 and periprosthetic valve leakage in 6 patients. At the initial operation, 10 patients underwent DeVega's tricuspid annuloplasty and 46 patients' tricuspid valves were normal. At the second operation, the surgical treatment of TR included tricuspid valve replacement (TVR) in 9 and tricuspid annuloplasty in 47. RESULTS: Two patients died postoperatively giving a 3.6% hospital mortality. The 54 survivors were followed up for 6-132 months (mean of 79.4 months). Heart function improved significantly in 8 with TVR and in 40 with tricuspid annuloplasty. Echocardiography showed moderate TR in 5 and severe TR in 1 patient with tricuspid annuloplasty who need a further surgical treatment. CONCLUSION: Pulmonary hypertension, myocardial dysfunction, and atrial fibrillation might be responsible for the development of late TR after left cardiac valve replacement. Tricuspid annuloplasty, as the surgical method of first choice, resulted in improvement in 87% of patients with late TR after left cardiac valve replacement. TVR can also be safely applied to repair organic disease and the extremely dilated tricuspid valve annulus. If the TR area is more than 25cm(2), the TVR is recommended. PMID- 16352172 TI - Follow-up of Australian aboriginal patients following open-heart surgery in Western Australia. AB - BACKGROUND: To date there has been only one published report pertaining to the outcomes following open-heart surgery in Australian aboriginal patients. METHODS: The records of 57 consecutive aboriginal patients who underwent open-heart surgery at our institution over a 6-year period were retrospectively reviewed. Attempts were made to contact the patient by telephone, by letter, through their listed next of kin, general practitioners, community nurse or through the aboriginal liaison officer. RESULTS: There were 38 males and 19 females. Forty five patients had coronary artery bypass grafting, 10 had isolated valve procedures while 2 had a combined operation. Mechanical prostheses were used for all valve replacements. There was a high incidence of diabetes, hypertension, hypercholesterolemia and smoking. There was one hospital death. Follow-up ranged from 6 months to 6 years with a mean of 3.09 years. Forty-four of the 56 (78.6%) hospital survivors were contactable while 21.4% were not contactable. There were two late deaths. Of the coronary patients who could be contacted, only 79% were taking Aspirin and a similar number Statin. 23% patients had recurrence of angina. There were five episodes of anticoagulation related complications in three patients. Only 44% of the patients were conversant with anticoagulation. CONCLUSIONS: The follow-up of the aboriginal patients was disappointing. The compliance with the medications was sub-optimal. There was a high incidence of recurrence of angina and anticoagulation related complications. The results raise concern about the use of mechanical prosthesis in these patients. PMID- 16352174 TI - Informed consent--legal issues "when yes really means no". PMID- 16352175 TI - The future of medical indemnity. PMID- 16352173 TI - The challenge of improving donor heart preservation. AB - Heart transplantation has in recent years become the treatment of choice for end stage heart failure. However while the waiting list for transplantation is growing steadily, the donor pool is not increasing. Therefore, in order to meet demand, transplant programs are using older, "marginal donors" and accepting longer ischaemic times for their donor hearts. As donor organs are injured as a consequence of brain death, during the period of donor management, at organ harvest, preservation, implantation and reperfusion, expansion of acceptance criteria places a great burden on achieving optimal long-term outcomes. However, at each step in the process of transplantation strategies can be employed to reduce the injury suffered by the donor organs. In this review, we set out what steps can be taken to improve the quality of donor organs. PMID- 16352176 TI - Bronchopulmonary sequestration supplied by the coronary circulation associated with a right-sided aortic arch. AB - Bronchopulmonary sequestration is an uncommon congenital anomaly. Although systemic arterial supply to the abnormal lung tissue is usually derived from the aorta, other nutrient vascular sources have also been described, including, infrequently, the coronary circulation. A right-sided aortic arch is another rare vascular abnormality, generally present in the setting of a vascular ring. We report a case of pulmonary sequestration receiving arterial supply from the right coronary artery, in the presence of a right-sided aortic arch, a constellation of anomalies not previously described. PMID- 16352177 TI - Retroaortic course of the anomalous left main coronary artery: is it a benign anomaly? A case report and review of literature. AB - BACKGROUND: Left coronary artery sharing a common origin with the right coronary artery and taking a retroaortic course is a rare coronary anomaly. It has been considered a benign entity. CASE REPORT: We report a 19-year-old man who presented with chest pain. Cardiac enzymes were elevated suggesting myocardial ischaemia. Coronary angiography revealed single origin of the coronary artery in the right coronary sinus. The left main coronary artery followed a retroaortic course before bifurcating. The anomalous left coronary artery was dissected and reimplanted in the left coronary sinus. The patient made an uneventful recovery. One year later, he is well with no recurrence of symptoms. CONCLUSIONS: Surgery to restore the normal anatomic position of the left coronary ostium is advisable whenever this anomaly is discovered. PMID- 16352178 TI - Mycotic coronary artery aneurysm detected by gallium67 scintigraphy. AB - A 71-year-old man presented with fever and positive blood cultures for methicillin-sensitive Staphylococcus aureus 4 days after an uncomplicated intra luminal abdominal aortic aneurysm repair. Investigations looking for a source of sepsis including computerized tomography scans, transoesophageal echocardiography, a bone scan and repeated chest X-rays, did not reveal an infective focus. Gallium(67) scintigraphy, however, showed a focus of tracer uptake in the region of the aorto-cardiac junction consistent with the presence of an abscess. Coronary angiography later opacified an aneurysmal cavity arising from the right sinus of Valsalva which was confirmed to be a mycotic aneurysm on thoracotomy. This is a rare case of early detection of a cardiac mycotic aneurysm by Gallium(67) scintigraphy. PMID- 16352179 TI - Enteritis-associated myocarditis. AB - We describe two cases in which myocarditis was associated with a prodromal diarrhoeal illness as a result of bacterial enteritis. Both cases were associated with elevated troponin T and chest pain mimicking myocardial infarction. PMID- 16352181 TI - Acute aortic dissection: how to confirm the diagnosis? PMID- 16352180 TI - Prostheses reforms. PMID- 16352182 TI - Outcomes of cardiac surgery in aboriginal Australians: what are the problems and what's to be done? PMID- 16352183 TI - Role of oxidative stress in cardiac remodelling after myocardial infarction. AB - Recovery from myocardial infarction is associated with a series of alterations in heart structure and function, collectively known as cardiac remodelling, which play a major role in the subsequent development of heart failure. Early remodelling involves infarct scar formation in the ischaemic zone whereas subsequent ventricular remodelling affects mainly the viable non-infarcted myocardium with especially profound alterations in the extracellular matrix. There is growing evidence for a role of oxidative stress and redox signalling in the processes underlying cardiac remodelling. Reactive oxygen species are a group of highly reactive molecules which have the potential to modulate several biological processes as well as cause tissue damage and dysfunction. Their effects can be beneficial or deleterious, depending on the concentrations produced, the site of production, and the overall redox status of the cell. Reactive oxygen species can be generated by all cardiovascular cell types. Under pathophysiological conditions, major enzymatic sources appear to be mitochondria, xanthine oxidase and the non-phagocytic NADPH oxidases. In this review, we outline the mechanisms underlying the progression of early and late cardiac remodelling with particular focus on the role of oxidative stress and the potential sources of reactive oxygen species which may be involved. PMID- 16352184 TI - Maze III replication using radiofrequency microbipolar coagulation. AB - BACKGROUND: The major concerns about the Cox's Maze III procedure have been the long bypass and cross-clamp times involved. We have developed a technique using microbipolar coagulation to duplicate most of the incisions described in the Maze III procedure. TECHNIQUE: This procedure is performed using standard hypothermic cardiopulmonary bypass and cold sanguineous hyperkalemic arrest. The left atrial Maze is performed using microbipolar coagulation following repair or following valve excision, prior to replacement. Cryolesions are used instead of radiofrequency at the mitral annulus to avoid injury to the circumflex coronary artery. Valve replacement is then done and the left atriotomies are closed. After rewarming, adequate deairing and removal of the aortic cross-clamp, the right atrial procedure is performed using microbipolar coagulation except ends of the radiofrequency lesion lines at the tricuspid annulus, where cryolesions are used. The rest of the surgery proceeds as usual. We used this procedure concomitantly with valve surgery in 84 patients. Sinus rhythm was restored in 86% of them. CONCLUSIONS: The use of radiofrequency energy in the microbipolar mode is safe and effective. The technique is easily reproducible and the tools involved are inexpensive and readily available. Although the cure rate is less compared to Cox's Maze III procedure and there is an attrition rate, sinus rhythm and atrial mechanical function is restored in the majority and with brief additional operative time. PMID- 16352185 TI - The Australian and New Zealand cardiac pacing and implantable cardioverter defibrillator survey: calendar year 2001. AB - AIM: A pacemaker (PM) and implantable cardioverter-defibrillator (ICD) survey was undertaken in Australia (Au) and New Zealand (NZ) for calendar year 2001. RESULTS AND CONCLUSIONS: Compared to the 1997 survey, significant increases in implantation numbers were recorded. For 2001, the total new PMs implanted was 9498 Au (6405 in 1997) and 914 NZ (823 in 1997). The number of new PM implants per million population was 486 Au (345 in 1997) and 245 NZ (228 in 1997). There were also significant increases in PM replacements between surveys with 1536 in Au (735 in 1997) and 195 in NZ (126 in 1997). Dual chamber implants were 71% Au (65% in 1997) and 56% NZ (55% in 1997). Pacing leads were overwhelmingly transvenous and bipolar with an increase in the use of active fixation leads in preference to tined leads, particularly in the atrium. There was a marked increase in the use of ICDs. The implants were 956 Au (449 in 1997) and 86 NZ (31 in 1997) with new implants per million population being 49 Au and 23 NZ. A breakdown of data for the six Au States and well as comparisons of similar surveys from other countries is presented. PMID- 16352186 TI - Saphenous vein grafts: to use or not to use? AB - The choice of the graft conduit is crucial to the success of coronary artery bypass grafting (CABG) because the patency of a coronary conduit is closely associated with an uneventful postoperative course and better long-term patient survival. From the beginning of coronary bypass surgery venous conduits particularly the great saphenous vein (GSV) has been the most frequently used coronary conduit. However, over the last decade or so, coronary bypass graft surgery with arterial revascularization of all diseased coronaries has shown to be efficient because arterial grafts have better long-term patency, especially left internal mammary artery (LIMA), compared with venous grafts. Early vein graft failure coupled with occlusion is the most important limitation of saphenous vein grafts. Nevertheless, vein grafting is still an integral part of cardiac surgical practice. This review provides a summary of the patency rates, technical features and certain characteristics of the venous conduits. It also examines the current understanding and knowledge of venous histology, vein graft pathology and the associated endothelial and smooth muscle cell physiology and pharmacology. In addition, the existing and the emerging strategies to combat and control vein graft intimal hyperplasia and accelerated atherosclerosis are reviewed in detail. PMID- 16352187 TI - Incidence of defibrillator patch infection due to pulmonary erosion. AB - BACKGROUND: Intrathoracic defibrillator patches were used prior to effective transvenous defibrillation systems. Spontaneous erosion through the visceral pleural has been documented but the incidence has not been previously reported. METHODS: All patients who underwent insertion of ICD patches at Westmead Hospital were identified from Hospital and device company databases. Medical records were reviewed and further follow-up performed through local doctors and the National Registry of Deaths where appropriate. RESULTS: One hundred and four patients underwent insertion of ICD intrathoracic patches between 1987 and 2000. The patches were positioned adjacent to the pericardium in the pleural cavity. Eight patients died within three months of surgery and two patients developed early ICD infection. Of the other 94 patients, 40 were alive at the time of last review such that there had been 562 patient years of survival with four patients developing spontaneous patch infection. The incidence was 0.7% per patient year. The diagnosis was delayed in all four patients. All underwent removal of the ICD system with one perioperative death. CONCLUSIONS: There is a small but significant risk of late spontaneous ICD patch infection. The condition is life threatening and the diagnosis is usually delayed. PMID- 16352188 TI - Homograft aortic valve replacement--the experience of one unit. AB - BACKGROUND: Homograft valves offer advantages including avoidance of anticoagulation and less susceptibility to infection especially in the setting of endocarditis. However, there is concern about their durability and possible accelerated degeneration particularly in cases of second time replacement with homografts. AIM: This study aimed to evaluate the pattern of homograft failure and the quality of life in patients after homograft implantation. METHODS: Between 1990 and 1998, 58 patients underwent aortic valve replacement with a homograft (aortic homograft = 47, pulmonary homograft = 11). Evaluation was based on clinical and echocardiographic examination, patient questionnaires and explanted valve pathology. Survival and freedom from cardiac related death were expressed by actuarial methods. RESULTS: Follow up ranged from 1 to 10 years (mean 5.5 years). Analysis of questionnaires revealed 60% of respondents to be in good performance status and 20% in moderate and 20% in poor performance status groups. Eleven patients (18.9%) required subsequent redo valve replacement after initial homograft insertion (pulmonary = 6, aortic = 5) due to either valve dehiscence (n=4) or valve degeneration (n=7). The mean interval of re-replacement was 5.4 years. CONCLUSIONS: Pulmonary homografts have a high failure rate in the aortic position. Overall subjective and clinical improvement after surgery is less than expected for a "physiological" device. In the setting of low availability of homografts the use of off-the-shelf devices such as stentless xenografts may be preferable in most cardiac surgical units in the current era. PMID- 16352190 TI - Activation of nuclear factor-kappaB and correlation with elevated plasma c reactive protein in patients with unstable angina. AB - BACKGROUND: Unstable coronary syndromes are currently believed to be caused by rupture of an atherosclerotic plaque due to local events, which may be of general inflammatory etiology. There is increasing evidence that nuclear factor-kappaB (NF-kappaB) is a key transcription factor in controlling gene expression concerning inflammatory response, and that plasma concentrations of C-reactive protein (CRP) is a sensitive marker of inflammation in unstable coronary syndromes. However, whether NF-kappaB activation is associated with coronary heart disease (CHD) activity as well as plasma CRP level has been less well investigated. The aim of this study was to explore whether NF-kappaB activation was associated with CHD activity and plasma CRP elevation in patients with unstable angina (UA). METHODS: NF-kappaB activity derived from white blood cells circulating in 33 patients with CHD was determined by electrophoretic mobility shift assay. Of these, 16 had UA and were within 24h of the last episode of chest pain. The remaining 17 were being evaluated for stable angina (SA). The CRP was also evaluated in both groups using a high-sensitivity ELISA. RESULTS: There was marked NF-kappaB activation and elevated levels of CRP in UA group compared with SA group (4.02+/-0.71 AU versus 1.24+/-0.23 AU and 5.0+/-0.7mg/l versus 1.4+/ 0.4mg/l, respectively, P<0.01), no NF-kappaB signal was observed in normal subjects (n=10). The NF-kappaB activation had a positive correlation with levels of CRP in patients with UA (n=11, gamma=0.771, P<0.01), but had no relationship between other clinical characteristics and the status of NF-kappaB activation. CONCLUSIONS: Our data suggest that inflammation is an important feature of unstable coronary artery disease, and both NF-kappaB and CRP are useful markers for the detection of UA or vulnerable plaques. PMID- 16352189 TI - Comparison of pulmonary gas exchange in OPCAB versus conventional CABG. AB - BACKGROUND: Cardiopulmonary bypass has been implicated as a cause of acute lung injury in cardiac surgical patients. This could be avoided with off-pump coronary artery bypass surgery. Aim. To ascertain the possible benefit of OPCAB surgery on pulmonary gas exchange. METHODS: We randomized 75 consecutive patients (mean age 57 years) into two groups: Group 1 off-pump coronary artery bypass surgery (OPCAB), n=37, Group 2 conventional coronary artery bypass grafting (con CABG), n=38. Alveolar-arterial oxygen difference (A-aO(2) difference) was calculated pre operatively, then 2 and 4h post-operatively. PaO(2)/FiO(2) ratio and respiratory index (RI) were calculated 2 and 4h post-operatively. RESULTS: Alveolar-arterial O(2) gradient sharply increased in the immediate post-operative period, from 27mmHg pre-operatively, to 227mmHg 2h post-operatively, then declined to 152mmHg 4h post-operatively. PaO(2)/FiO(2) ratio and RI also showed severe worsening 2h post-operatively, with marked improvement at 4h. The pattern of physiological deterioration of gas exchange was similar in both the groups. CONCLUSION: In terms of pulmonary gas exchange, similar degree of deterioration is noticed in CABG patients with or without cardiopulmonary bypass. OPCAB seems to provide no physiological benefit of gas exchange at the alveolar capillary membrane when compared to conventional CABG. PMID- 16352192 TI - False positive computed tomography findings in aortic dissection. AB - Dissection is the commonest acute catastrophe involving the aorta. Computed tomography (CT) is increasingly used in evaluating patients with suspected aortic dissection. Occasionally normal anatomic structures may be misconstrued as pathologic due of lack of familiarity with anatomical variations. In scanning two patients with suspected dissection, we encountered pitfalls that led to an erroneous diagnosis of dissection. The subsequent clinical course and evaluation confirmed that the CT findings were artefactual. This report illustrates these potential errors and discusses the mechanisms. PMID- 16352191 TI - Different regulation of ERK1/2 activation by beta-adrenergic receptor subtypes in adult mouse cardiomyocytes. AB - BACKGROUND: Increasing evidence suggests that stimulation of beta-adrenergic receptor (betaAR) activates mitogen-activated protein kinases (MAPKs), particularly extracellular signal-regulated kinase (ERK1/2) which is involved in the regulation of a multitude of cellular processes. However, the subtype specific effects of betaAR stimulation on MAPKs remain to be elucidated. AIMS: In the present study, we determined whether beta(1)AR and beta(2)AR differ in regulating ERK1/2 activation in the myocardium. METHODS: To avoid complicated interactions between betaAR subtypes, we separately expressed either beta(1)AR or beta(2)AR using adenoviral gene transfer in adult mouse cardiac myocytes from beta(1)beta(2) double knockout mice. RESULTS: Stimulation of beta(1)AR by isoproterenol markedly increased ERK phosphorylation and activity by 2.1-fold in a time-dependent manner. In contrast, stimulation of beta(2)AR slightly decreased ERK activation. Furthermore, pretreatment of cells with pertussis toxin to disrupt Gi function did not affect the inhibitory effect of beta(2)AR on ERK1/2. CONCLUSIONS: We have shown that stimulation of cardiac betaAR subtypes differentially regulates ERK activation in adult mouse cardiomyocytes. PMID- 16352193 TI - A case of Fabry disease mimicking hypertrophic cardiomyopathy. PMID- 16352194 TI - Thymoma presenting with infarction. AB - Thymomas are the most common tumours of the anterior mediastinum with most patients presenting incidentally. We present a case of thymoma presenting with sudden onset severe chest pain and raised plasma aminotransaminases secondarily to spontaneous infarction. We discuss the presence of these aminotransaminases in this highly unusual presentation of thymoma and believe our case demonstrates the inherent limitations of plasma enzymes due to their lack of specificity and sensitivity. PMID- 16352195 TI - Transcatheter coil occlusion of coronary artery to right ventricular fistulae in pulmonary atresia with intact ventricular septum. AB - We describe a young adult with an underlying diagnosis of pulmonary atresia with intact ventricular septum and Fontan physiology who presented with extreme dyspnoea on exertion. Coronary artery to right ventricular fistulae and reversible myocardial ischaemia secondary to a right ventricular steal phenomenon were present. The right coronary artery to right ventricular fistula was completely occluded with coils, by a transcoronary approach, with amelioration of symptoms. However, one coil embolised and lodged uneventfully in the left atrium during the procedure. This case demonstrates that a transcatheter approach may be successfully employed not only in patients with isolated coronary artery fistulae, but also in patients with coexistent complex congenital heart disease. PMID- 16352196 TI - The correct table height. PMID- 16352197 TI - Acute stent thrombosis associated with exercise testing. PMID- 16352198 TI - Towards stroke-free coronary surgery. PMID- 16352199 TI - Surgery for atrial fibrillation: putting the pieces of the jigsaw together. PMID- 16352200 TI - From measurement to improvement in cardiac surgical care. PMID- 16352201 TI - Warfarin and atrial fibrillation. PMID- 16352202 TI - Should doctors discourage nutritional supplementation? A cardiovascular perspective. AB - Cardiovascular disease is the major cause of death in most western countries, and current preventive treatment is relatively ineffective. There are theoretical reasons why nutritional therapy-antioxidant vitamins, minerals and omega 3 oils should be beneficial. Because of some negative trials, there is the risk of tossing the baby out with the bath water. This paper reviews how nutritional therapy could assist in a comprehensive preventive approach to reduce heart disease-especially primary prevention. It stresses the need to separate primary prevention-initiated by oxidation of LDL in the vessel wall, and secondary prevention-the consequences of plaque rupture. Because a therapy is ineffective in the secondary preventive role, does not mean that it could not have a primary preventive action. There are many positive studies utilizing nutritional therapies, these are detailed in this paper, and a comprehensive preventive programme including these is suggested. PMID- 16352203 TI - Atrial fibrillation, stroke and anticoagulant use. AB - BACKGROUND: Ischaemic stroke is reduced by anticoagulant therapy in high-risk patients with atrial fibrillation. Evidence however, suggests patients are under treated. AIM: To assess anticoagulant use in patients with ischaemic stroke and atrial fibrillation in Waikato Hospital. METHODS: A retrospective review of all patients admitted with stroke over a one-year period. RESULTS: Ischaemic stroke occurred in 189 patients with atrial fibrillation noted in 21% (39/189) of this group. The majority were female, 59% (23/39) with a mean age of 79.8 years. A total of 84% (33/39) were considered high risk for thromboembolic complications but only 18% (6/33) were anticoagulated prior to the stroke. Following the cerebrovascular accident all patients were considered to be at high risk but only a further 25% (10/39), were anticoagulated. In the remaining 55% (21/39) anticoagulant therapy was not commenced with age alone cited as a contraindication in 8% (3/39). At a mean follow-up of 10 months no complications of anticoagulant therapy were reported. CONCLUSIONS: A significant proportion of patients with atrial fibrillation and high-risk characteristics are not anticoagulated prior to ischaemic stroke. These findings reflect overseas experience. Reasons are unknown but may in part relate to physician reluctance to anticoagulate elderly patients. PMID- 16352204 TI - Factors affecting diurnal variability of ventricular tachyarrhythmias detected by multiprogrammable implantable cardioverter-defibrillators. AB - Many cardiovascular events, including ventricular arrhythmias, display diurnal variability with a morning peak, and a less pronounced afternoon peak. Since the advent of multiprogrammable implantable cardioverter-defibrillators (ICDs), it has been possible to analyse ventricular tachyarrhythmic events. This study aims to evaluate the circadian pattern of ventricular tachycardias in patients treated with ICDs and examines whether antiarrhythmic medications affect this pattern. Data recorded from 83 patients' ICDs were manually analysed and events other than ventricular arrhythmias were excluded. There was a morning peak of ventricular arrhythmias at around 9.00 a.m. This peak was maintained in patients with ejection fractions of less than 40% and those whose arrhythmias had cycle lengths of less than 230ms. Beta blockers appeared to have no effect on this morning peak but the peak appeared later with amiodarone. PMID- 16352205 TI - Counselling strategies in the prenatal diagnosis of major heart abnormality. AB - BACKGROUND: Prenatal diagnosis of a major cardiac abnormality precipitates a crisis in the parents. To their grief and/or anger may be added the burden of whether to continue with the pregnancy if the fetus is previable. AIMS: We reviewed our counselling strategies to determine if the parents were satisfied with our approach and whether in their perception it provided the knowledge and understanding of the abnormality detected to help them in their decision. METHODS: Questionnaires highlighting parental perception of their experiences, were developed and sent to a cohort of parents following the birth of their infant or at least 6 months after termination. The parents were seen over 3 years in a private obstetric ultrasound practice, diagnosed with or specifically referred because of the diagnosis of a major heart abnormality in the fetus, confirmed at a subsequent scan with the paediatric cardiologist in attendance. Both clinicians explained the abnormality, 'working off each other'. The obstetrician ensured the explanations were in lay terms. Anatomical detail, while important to the clinicians, was not emphasised, but rather function, quality of life issues and probable outcome and attendant risks of intervention. An appointment with the cardiologist was offered as was the need/option for further scans. RESULTS: 39 patients were reviewed, 13 of whom elected to terminate their pregnancy. Of the 30 who responded, 70% reported that the explanations of the abnormality were clear to very clear, whilst 85% considered the detail given was 'just right'. Only 60% rated their understanding of the diagnosis as 'good to excellent', improving to 80% with subsequent scans. These results were especially meaningful when 70% of the respondents described themselves as being distressed or very distressed following the initial diagnosis. CONCLUSIONS: These results suggest that parents seemed satisfied with our approach of two clinicians working together emphasising function and outcome, rather than anatomical detail. Their perceived understanding seemed to allow them to make their decision concerning continuation or termination of their pregnancy. PMID- 16352206 TI - Uncovering a hidden epidemic: a study of the current burden of heart failure in Australia. AB - BACKGROUND: Australia, like other countries, is experiencing an epidemic of heart failure (HF). However, given the lack of national and population-based datasets collating detailed cardiovascular-specific morbidity and mortality outcomes, quantifying the specific burden imposed by HF has been difficult. METHODS: Australian Bureau of Statistics (ABS data) for the year 2000 were used in combination with contemporary, well-validated population-based epidemiologic data to estimate the number of individuals with symptomatic and asymptomatic HF related to both preserved (diastolic dysfunction) and impaired left ventricular systolic (dys)function (LVSD) and rates of HF-related hospitalisation. RESULTS: In 2000, we estimate that around 325,000 Australians (58% male) had symptomatic HF associated with both LVSD and diastolic dysfunction and an additional 214,000 with asymptomatic LVSD. 140,000 (26%) live in rural and remote regions, distal to specialist health care services. There was an estimated 22,000 incidents of admissions for congestive heart failure and approximately 100,000 admissions associated with this syndrome overall. CONCLUSION: Australia is in the midst of a HF epidemic that continues to grow. Overall, it probably contributes to over 1.4 million days of hospitalization at a cost of more than 1 billion dollars. A national response to further quantify and address this enormous health problem is required. PMID- 16352207 TI - Renal artery stenting for renal artery stenosis: interventional approach and initial clinical results. AB - BACKGROUND: Significant renal artery stenosis is being increasingly diagnosed and interventional cardiologists are taking on the unfamiliar role of deploying renal stents. We describe our technique for renal artery stenting using techniques and equipment similar to that used in coronary stenting and present early results. METHODS: Thirty-two patients with significant renal artery stenosis (diameter stenosis 60-95%) and difficult to control hypertension or renal impairment were referred for renal stenting. Renal DTPA scans were performed before intervention. All the procedures were attempted from the femoral artery approach using guides, low profile wires and balloon catheters similar to those used for coronary work. The lowest profile renal or coronary stents were deployed using a monorail technique. RESULTS: Atherosclerosis was the cause of the renal artery stenosis in 31 patients. Stents were successfully deployed in 30 of these patients (97%). One patient required a second procedure from the brachial artery approach. An additional patient with fibromuscular dysplasia underwent angioplasty only. Angiographic diameter stenosis was reduced from a mean of 79 to 1%. Translesional mean pressure gradients were reduced from 22 to 1mmHg. There were no major procedural or in-hospital complications. Anti-hypertensive drugs were reduced from a mean of 3 prior to stenting to 0.7 at discharge. Renal DTPA scans were a poor predictor of severity of stenosis. Renal DTPA scans suggested functionally significant renal artery stenosis in only 48% of patients but the clinical responses suggested that this was an underestimate. CONCLUSIONS: Stenting of renal arteries using coronary like techniques and equipment results in a high rate of procedural success and good early clinical outcomes. PMID- 16352208 TI - Surgical treatment of atrial fibrillation. AB - This article reviews the fundamentals of surgical treatment of atrial fibrillation. The clinical classification, pathophysiology, medical treatment strategy, and catheter-based interventions are also included. The Cox-Maze III procedure was developed over based on experiences of several operations which proceeded it. The operation is complex but results are excellent with over 90% of patients in normal sinus rhythm or regular atrial rhythm. Several modifications have been devised using various energies to ablate atrial myocardium as a means of extending surgical incisions to simplify the Maze III operation. Techniques and results of these operations are reviewed. Modified operations offering about 80% restoration of sinus rhythm are attractive to more surgeons because of the reduced complexity and time of operation. PMID- 16352209 TI - Vettath's anastamotic obturator--our experience of 269 proximal anastomoses. AB - BACKGROUND: Coronary artery bypass grafting (CABG) has come full circle-it started as an off-pump affair, then became an on pump one and now we are trying to keep off the pump again. One of the main reasons for this has been the neurological sequelae subsequent to CABG. But neurological problems kept causing concern even in off-pump CABGs (OPCAB). Side clamping the aorta was thought to be the major factor and thus came the concept of 'no touch proximal anastomoses' onto the aorta (1). Though a variety of proximal anastamotic devices are available in the market, high cost is a matter of real concern in third world countries like India. Hence, this endeavor of ours to fabricate an anastamotic device of our own-'the Vettath's anastamotic obturator' (VAO) for proximal anastomoses of saphenous vein grafts (SVG) onto the aorta. VAO is a stainless steel rod with three grooves and a guard at the end, which sinks into the aorta, through a punch hole, cordoned off by two wide purse string sutures. METHODS: After trials on perfused animal heart models, we started using this device on humans. We have performed 269 proximal anastomoses using the VAO in 177 of our OPCAB patients in the past 1 year (till July 2003). Ninety-five of them had single top ends, 72 had 2 top ends and 10 had 3 top ends onto the aorta. We have used this on disease free islands on four patients with palpable aortic plaques. Initially all anastomoses were of the proximal first type (to ensure that the flow was adequate). Now-a-days, with confidence, distal first anastomoses are being performed. RESULTS: We had no operative mortality in this group. None of our patients needed IABP support. One patient reported back with angina, after 3 months-he was studied and his grafts were found to be patent. All patients, except three, are being followed up till date and they are leading active symptom free and event free lives. DISCUSSION: It is logical to think that avoidance of side clamp on the aorta reduces the risk of neurologic complications. Vettath's anastamotic obturator is an indigenous, cheap and reusable alternative to the other costlier devices, which serve the same purpose. Though there is a small learning curve, results are gratifying and complications are few. PMID- 16352210 TI - Initial twelve months experience and analysis for 2001-2002 from the Australasian Society of Cardiac and Thoracic Surgeons--Victorian database project. AB - The Australasian Society of Cardiac and Thoracic Surgeons (ASCTS) have established a database for the collection and analysis of the results of cardiac surgery in Australia and New Zealand. Initially data has been collected only in Victoria public hospitals. This report covers the first 12 months of data collection from 1st August 2001 to 1st July 2002. BACKGROUND: Whilst cardiac surgical performance in Australia is considered to be of a high standard equivalent to other developed countries, there is currently no systematic approach to data collection in order to provide performance indicators and benchmarks. The development of an Australasian cardiac surgical database and performance indicators will enable benchmarking and comparison with international standards which should lead to performance improvements. METHODS: A database definition set and standardised data collection form was developed by the ASCTS for all participating cardiac surgery units in public hospitals in Victoria. Opt off consent for subject inclusion in the database was approved by each participating institutional ethics review committee. An electronic database and reporting application was developed. Data included in this analysis is from the initial 12 months collection from all hospitals participating in the project from 1st August 2001 to 31st July 2002. RESULTS: Overall, there were 2982 procedures performed in this period of which 2969 had sufficient data to be included in this analysis (99.5%). The majority of procedures 2017 (68%) being undertaken were isolated coronary artery bypass surgery (CABG). The mean age of all subjects undergoing procedures was 65 years (range: 18-91 years) and 70% were male. 64% of all procedures were elective and 6.1% emergency or salvage. Median post-procedure length of hospital stay for all procedures was 6.0 days and intensive care unit (ICU) stay was 23.0h. Re-operation for haemorrhage occurred in 2.1% of all cases and deep sternal infection in 0.4% of all cases. Crude 30-day operative mortality was 3.6% for all procedures; 2.1% for isolated CABG, 3.6% for valve procedures, 5.2% for valve and CABG and 11.4% for other cardiac surgical procedures. Mortality rates increased from 1.8% for elective procedures to 4.1% for urgent and 24.6% for emergency or salvage operations. In comparison to international figures from the USA and UK, mortality rates following isolated CABG were lower whilst average length of hospital stay post-procedure was higher. CONCLUSION: The ASCTS database project is now well established and the electronic database and reporting module is in operation in all participating sites. The risk-adjusted isolated operative mortality suggests cardiac surgical performance in Victoria compares well with international standards. As the database develops, local risk adjustment models for mortality and morbidity for each procedure will be developed to enable appropriate between hospital comparisons. PMID- 16352211 TI - Early audit of renal complications in a new cardiac surgery service in Australia. AB - OBJECTIVE: To assess the incidence of renal failure in a cardiac surgery service commencing in Australia. DESIGN: Prospective data collection and retrospective database analysis. SETTING: A tertiary referral, university teaching hospital in the state of Victoria, Australia. PARTICIPANTS: The first 502 patients undergoing cardiac surgery in this institution from commencement of the service. RESULTS: The overall rate of renal failure was low in comparison to other studies at 0.2% (95% CI 0.04-1.3%). The rate of postoperative renal dysfunction was also low at 4.2% (95% CI 2.7-6.5%). CONCLUSIONS: The safety of the new service with respect to this complication of cardiac surgery was good when compared with published data. However the lack of uniform definitions of renal failure following cardiac surgery make comparisons between studies difficult. Uniform reporting of this complication would facilitate comparisons between units and quality assurance activities in this field. PMID- 16352212 TI - Ensuring transmurality using irrigated radiofrequency modified maze in surgery for atrial fibrillation--a simple and effective way. AB - Transmural lesions are ensured with the cut and sew technique and with bipolar delivery of radiofrequency (RF) energy [Ann. Thorac. Surg. 74 (2002) 2210]. Unipolar sources of energy such as radiofrequency and microwave do not guarantee transmurality and may require intraoperative testing to evaluate complete lesion depth. Transmural lesions are generally effective electrophysiological lesions and are desirable as it does not allow atrial fibrillation (AF) breakthrough or development of atrial flutter in most instances. We describe a simple and effective way of ensuring transmurality using irrigated radiofrequency in the modified maze surgery for AF. PMID- 16352213 TI - A complicated percutaneous coil embolisation of a large coronary artery fistula. AB - A coronary artery fistula is a rare cause of cardiac symptoms or signs in an adult. We report here the successful percutaneous treatment of a large fistula in an adult who had suffered an acute myocardial infarction peri-operatively due to a steal phenomenon. Due to the fistula size, a specially constructed occlusion device was utilised for effective treatment. PMID- 16352214 TI - Successful repair of a rare angiographic presentation of coronary artery-coronary sinus fistula. AB - We report a rare case of right coronary artery-coronary sinus fistula successfully closed through coronary sinus in the second operation following identification of distal branches of the coronary artery. At nine months follow up, patient is asymptomatic with no recurrence of cardiac murmur. Repeat coronary angiography at one year showed no connection between right coronary artery and coronary sinus. This case report highlights the difficulties in angiographic techniques, identification and interpretation of anatomy of these rare malformations. It also demonstrates the hazards of simple ligation of these anomalous vessels without first understanding the anatomy of the regional coronary circulation. A brief review of literature on coronary artery fistula will be presented. PMID- 16352215 TI - Coronary artery fistula in a patient with pulmonary atresia and tricuspid atresia clinical and MRI findings. AB - The MRI findings of a case of coronary artery fistula occurring in a patient with pulmonary atresia and tricuspid atresia is presented. PMID- 16352216 TI - A Cardiac Clinico-Pathological Conference in 1882 an historical vignette. AB - In the Australian Medical Journal of March 1883, there is a report entitled, "Valvular incompetency of pulmonary artery, with regurgitation", presented by Dr. William Pegus. Dr. Pegus presented this paper about his own peculiar heart problem, which was different from any in the textbooks of that time, to a group of doctors in 1882. He described his symptoms, the progress of the condition and the treatment he received, the most helpful of which was the use of Turkish baths. There was discussion on the possible diagnosis but at the conclusion of the presentation it was felt that all the symptoms could not be explained and therefore the diagnosis was uncertain. In the light of present day knowledge, the clinical picture is typical of a rupture of aneurysm of a sinus of Valsalva into the right ventricle. Rupture of such an aneurysm was first described by Hope in 1839 in his book on diseases of the heart and great vessels, the diagnosis being made at autopsy. Dr. Pegus was probably the first person in Australia to publish a detailed description of the symptoms and clinical signs and to give an account of the progress of the condition with his own thoughts about the treatment available at the time. PMID- 16352217 TI - Putting the heart through its paces-cellular insights from frequency phenomena. PMID- 16352218 TI - Nutritional supplements and cardiovascular disease. PMID- 16352219 TI - Myocardial tissue velocities in the normal left and right ventricle: relationships and predictors. AB - OBJECTIVES: (1) To document, using tissue Doppler echocardiography (TDE), the regional variations in myocardial velocities (MV) in the left and right ventricles (LV and RV) in healthy young adults. (2) To determine the factors predicting MV. BACKGROUND: The pattern of tissue velocities within the right ventricle have yet to be determined and their patterns compared to the left ventricular velocities have not yet been described. METHODS: Forty healthy subjects, mean age 29+/-6 years, were studied using TDE. Left ventricular long axis velocities (V(LV-LX)) were obtained by sampling from anteroseptal, anterior, lateral, posterior, inferior and inferoseptal LV walls, and long-axis RV velocities (V(RV-LX)) from the free wall of the RV, in standard apical views. LV radial velocities (V(LV-RAD)) and RV radial velocities (V(RV-RAD)) were assessed from the parasternal long and short-axis views. Regression analyses were performed to assess for correlations of MV with the variables: age, sex, QRS duration, heart rate, systolic and diastolic blood pressure, LV mass, width, LV or RV lengths, LA or RA areas. RESULTS: There were marked but consistent regional variations in systolic and diastolic tissue velocities in the LV and RV. Systolic (S') and early diastolic (E') velocities differed significantly around the left ventricular base, the highest velocities being located within the free wall at 6.4+/-2.2cm/s and 11.3+/-3.1cm/s, respectively. The E'/S'ratio remained constant and independent of position. V(LV-LX) were significantly higher than V(LV-RAD) (p<0.001). V(LV-LX)S' velocities were consistently lower than V(RV-LX)S' velocities (p<0.001). Age, heart rate, LV mass, width and length were significantly and independently associated with V(LV-LX)S' and V(LV-LX)E' values (p<0.01 for each). CONCLUSIONS: In healthy young adults, there is a consistent pattern of non-uniform MV throughout the heart, including differences in longitudinal and radial axis velocities both within the LV and between the LV and RV. Age, heart rate and LV structure are important determinants of MV. CONDENSED ABSTRACT: The patterns of left and right ventricular myocardial velocities and their relationships to each other are not well characterized. Furthermore, the determinants of myocardial velocities are not known. This study evaluated the myocardial longitudinal and radial axis tissue velocities in both the left (LV) and right (RV) ventricles and found that a consistent but non-uniform relationship exists between the LV and RV in both longitudinal and radial axes. Furthermore, age, heart rate and LV dimensions account for between 20% and 70% of the variability seen in LV systolic and diastolic velocities. PMID- 16352220 TI - Hand-assisted Thoracoscopic Surgery causes less postoperative pain than limited thoracotomy after cessation of epidural analgesia. AB - BACKGROUND: Hand-assisted Thoracoscopic Surgery (HATS) is a novel minimally invasive technique for performing procedures conventionally performed by posterolateral thoracotomy. HATS overcomes a major drawback of thoracoscopic surgery in allowing full manual palpation of the lungs via a subcostal (mini Kocher's) incision under videoscopic guidance, avoiding a thoracotomy, when the indication is pulmonary metastasectomy with curative intent or resection of undiagnosed lung nodules. It is postulated HATS may produce improved postoperative quality of life outcomes compared to thoracotomy. AIMS: To determine if HATS compared to limited posterolateral thoracotomy causes less postoperative pain after cessation of epidural anaesthesia. METHODS: Fifty-two patients were prospectively randomised to receive a limited thoracotomy (n=26) or HATS (n=26). Pain scores, measured on a visual analogue scale, were recorded during standard nursing observations and after analgesia demands in the 24h after epidural removal. RESULTS: Pain scores were significantly lower after HATS compared to limited thoracotomy (3.8 versus 5.2, p=0.04). There was no difference in postoperative respiratory function. CONCLUSION: HATS results in lower postoperative pain after cessation of epidural analgesia. This form of analgesia may therefore not be required, reducing the management complexity, complications and hospital stay associated with its use. SHORT ABSTRACT: Hand-assisted Thoracoscopic Surgery (HATS) is a novel technique allowing full manual lung palpation as an adjunct to Video-assisted Thoracoscopic Surgery (VATS). Fifty-two patients were prospectively randomised to receive limited thoracotomy or HATS. Pain scores were significantly lower after HATS compared to thoracotomy, indicating epidural analgesia may not be required. PMID- 16352221 TI - Patency of the radial artery graft: angiographic study in 209 symptomatic patients operated between 1995 and 2002 and review of the current literature. AB - OBJECTIVES: The purpose of this study is to define the long-term patency of the radial artery (RA) graft and review the current literature. METHODS: Two hundred and eighty-six RA symptom-directed graft angiograms were studied in 209 patients. The preoperative patient characteristics and intraoperative variables were collected prospectively from patients who had primary coronary artery bypass grafting between 1995 and 2002. A total of 166 (79%) patients were male with a mean age of 65 years. The mean period from operation to re-angiogram was 35 months. Actuarial techniques are not valid in graft patency studies as the time when the graft occluded is not known. Therefore, RA patency was analyzed at four categorical time intervals. The RA was grafted to the left anterior descending artery (LAD) in six patients (2%), diagonal (DIAG) in 29 (10%), obtuse marginal (OM) in 166 (58%), right coronary artery (RCA) in 9 (3%) and posterior descending artery (PDA) in 76 (27%) cases. The graft failure was defined as >or=80% stenosis. RESULTS: A total of 259 (91%) grafts were patent and 26 (9%) had failed. Most grafts were widely patent or occluded. The LAD/DIAG patency was 30/35 (86%), OM patency 154/166 (93%) and RCA/PDA patency 79/84 (94%). The interval from surgery to angiogram did not affect the RA graft patency (86% at <1 year, 95% at 1-3 years, 89% at 4-5 years, 96% at >5 years). CONCLUSIONS: Even in a patient cohort with adverse symptoms, excellent RA patency was achieved that remained almost constant through all time intervals studied. Better selection, harvesting and preservation may further improve early patency. PMID- 16352222 TI - Predictive value of EuroSCORE and Parsonnet scoring in Saudi population. AB - BACKGROUND: There is no previously published report on the applicability of risk stratification systems to adult cardiac patients in Saudi Arabia. AIM: We have tried to verify the predictive value of Parsonnet and EuroSCORE in Saudi patients. METHODS: Data of 194 consecutive adult patients operated in our unit over the period of last few years, was obtained retrospectively. There were no omissions. The EuroSCORE and Parsonnet scores were estimated for all the patients and compared with the observed mortality, morbidity and hospital length of stay (LOS). Calibration was determined by Hosmer-Lemeshow goodness of fit statistics (HL) while discrimination was assessed by using receiver operating characteristic (ROC) curve. RESULTS: For prediction of "mortality" the area under ROC curve was 0.766+/-0.077 S.E. for EuroSCORE and 0.685+/-0.087 S.E. for Parsonnet score. For "morbidity" the area under ROC curve was 0.628+/-0.048 S.E. for EuroSCORE and 0.660+/-0.048 S.E. for Parsonnet score while for the LOS, ROC curves were 0.632+/ 0.056 S.E. and 0.671+/-0.053 S.E. accordingly. Both scoring systems were adequately calibrated in our data. CONCLUSION: Parsonnet and EuroSCORE are equally applicable as mortality predictors in Saudi population. EuroSCORE was slightly better than Parsonnet scoring in predicting the mortality in medium and high risks categories. Both the systems have less discrimination for prediction of "morbidity" and LOS. PMID- 16352223 TI - Biatrial or bicaval technique for orthotopic heart transplantation: which is better? AB - BACKGROUND: Orthotopic heart transplantation was done by the biatrial technique initially and the bicaval technique has become popular recently. AIMS: This study aims to determine if bicaval technique is advantageous. METHODS: Consecutive transplants performed between 1996 and 2001 were divided into two groups-37 patients done by bicaval and 38 by biatrial technique. Data accumulation was by retrospective study of patient charts. RESULTS: Both groups had similar preoperative variables. There were no differences in low cardiac output (18.9% versus 26.3%, p = 0.62), intraaortic balloon pump insertion (16.2% versus 15.7%, p = 1.0), re-exploration (13.5% versus 18.4%, p = 0.79) and perioperative mortality (5.4% versus 7.9%, p = 1.0) in the bicaval versus biatrial groups. Temporary (13.5% versus 39.4%, p = 0.15) and permanent pacing (0 versus 3 patients) tended to be less frequent and central venous pressure measured at 1 week was lower in the bicaval group (mean 13.8 +/- 6cm versus 14.9 +/- 5.4cm, p = 0.42), but not attaining statistical significance. Severe tricuspid regurgitation was seen in one bicaval versus five biatrial patients at follow-up. CONCLUSIONS: Though bicaval group tended to require less pacing, had less tricuspid regurgitation and had lower central venous pressures, these did not attain statistical significance. There were otherwise no obvious differences in outcome. SHORT ABSTRACT: Seventy five consecutive orthotopic heart transplantations done during the period 1996-2001 by bicaval or biatrial surgical technique were compared. There was no difference in low cardiac output, intraaortic balloon pump insertion and mortality but the bicaval patients tended to have less pacing and diuretic requirements and lower central venous pressures, though not attaining statistical significance. PMID- 16352224 TI - Skeletonized bilateral internal mammary arteries for total arterial myocardial revascularization. AB - BACKGROUND: Bilateral internal mammary artery (IMA) grafting is associated with an improved long-term survival, low rates of recurrence of angina and late myocardial infarction. However, because of the inadequate length of the conduit, use of bilateral internal thoracic artery grafting occasionally is not suitable for complete revascularization. To overcome this limitation, extra length can be obtained by skeletonization of both IMAs. We decided to prospectively assess the safety of this technique. METHODS: One hundred patients with a mean age of 52.5 +/- 13.1 years underwent complete revascularization with skeletonized bilateral internal mammary arteries on cardiopulmonary bypass (CPB). The right internal mammary artery (RIMA) was used as a free graft connected to the in situ left IMA (LIMA) in 88 patients. A free LIMA was attached to in situ RIMA in 12 patients. The average number of grafts was 3.2 per patient (range: 2-4 grafts per patient). Mean left ventricular ejection fraction was 60% (range: 25-80%). RESULTS: No patient required reexploration for bleeding, and no patient died within 30 days after operation. On the basis of electrocardiographic changes, three patients sustained a perioperative myocardial infarction. One patient had a sternal wound infection. Mean follow-up was 24 months (range: 6-36 months). The actuarial survival rate was 99 +/- 1% at 3 years. No myocardial infarctions were reported during the follow-up. Three patients had recurrent angina with conduit occlusion diagnosed on coronary angiography. CONCLUSION: Complete myocardial revascularization with skeletonized bilateral internal mammary arteries is a safe and reliable technique with excellent early and mid-term results. PMID- 16352225 TI - Muscle and myocutaneous flaps in reconstructive surgery of thoracic defects. AB - Reconstructive surgery of thoracic defects presents a challenge for the surgeon. With defects of different aetiology and the need for precise localisation of the area to be treated, a broad range of experience is required. We present our interdisciplinary experience in dealing with full thickness thoracic wall defects and intrathoracic cavities. The latissimus dorsi muscle as well as the pectoralis major muscle and their covering skin are the most commonly used flaps in covering an intrathoracic or extrathoracic defect. They have the advantage of being easily and safely dissected. Other flaps such as the greater omentum, serratus anterior, the transverse rectus abodominal muscle (TRAM), and the filet of the arm are less frequently used. Indications and applications of these flaps are reviewed. Our interdisciplinary surgical treatment of thoracic wall defects allows optimal operative excision and reconstruction as well as giving best functional and aesthetic results for the patients. PMID- 16352226 TI - Saphenous vein grafts: to use or not to use? AB - The choice of the graft conduit is crucial to the success of coronary artery bypass grafting (CABG) because the patency of a coronary conduit is closely associated with an uneventful postoperative course and better long-term patient survival. From the beginning of coronary bypass surgery venous conduits particularly the great saphenous veins (GSV) have been the most frequently used coronary conduit from the beginning of the coronary bypass surgery. However, over the last decade or so, coronary bypass graft surgery with arterial revascularization of all diseased coronaries has shown to be efficient because arterial grafts have better long-term patency, especially left internal mammary artery (LIMA), compared with venous grafts. Early vein graft failure coupled with occlusion is the most important limitation of saphenous vein grafts. Nevertheless, vein grafting is still an integral part of cardiac surgical practice. This review provides a summary of the patency rates, technical features and certain characteristics of the venous conduits. It also examines the current understanding and knowledge of venous histology, vein graft pathology and the associated endothelial and smooth muscle cell physiology and pharmacology. In addition, the existing and the emerging strategies to combat and control vein graft intimal hyperplasia and accelerated atherosclerosis are reviewed in detail. PMID- 16352228 TI - A case of Coronary-Subclavian Steal Syndrome. AB - Surgical bypass of left coronary artery disease using the internal mammary artery (IMA) as a conduit is standard practice. Adequate blood supply to the IMA is dependent on normal flow in the subclavian artery. Stenosis of the subclavian can impair coronary supply from the internal mammary artery. This may create a steal syndrome whereby the left arm may rob the coronary system of flow especially during arm exercise. In the present report a case of angina with left coronary system steal via an internal mammary graft due to atherosclerotic subclavian stenosis. Successful treatment of the anginal symptoms by native coronary stenting is described. PMID- 16352227 TI - Pacing staircase phenomenon in the heart: from Bodwitch to the XXI century. AB - The frequency of pacing is a fundamental physiological modulator of myocardial function. When the pacing rate increases there is normally an increase in contractility (a positive force-frequency relationship). However in small rodents, fish and end-stage failing myocardium, the force-frequency response has been found to be flat or even negative. The positive staircase is understood to be related with the increase in the intracellular Ca(2+) transient, mainly due to an enhanced sarcoplasmic reticulum Ca(2+) content at higher stimulation frequencies, resulting from an increase in Ca(2+) influx per unit time and reduced Ca(2+) efflux between beats. However, additional mechanisms, such as increased activity of Ca(2+)/calmodulin-dependent protein kinase or enhanced myofilament responsiveness to Ca(2+) may also play a role. Although an increase in contraction frequency has been shown to be associated with an increase in intracellular Na(+), several studies have shown a temporal dissociation between the increase in Na(i)(+) and the increase in force evoked by changes in pacing frequency. The way in which the Na(+)/Ca(2+) exchanger contributes to contraction frequency inotropy is still not well understood. The aim of this review is to examine the contribution of the fundamental components of cardiac excitation contraction coupling to frequency inotropy in healthy and failing hearts. PMID- 16352229 TI - Conservative management of a large oesophageal haematoma as a complication of systemic thrombolysis. AB - An acute coronary syndrome patient was treated with tissue plasminogen activator to produce thrombolysis. Six hours post-lysis, haematemesis occurred. Computed tomography (CT) revealed a large haematoma around the lower oesophagus and endoscopy showed a tear in the lower end of the oesophagus. This case represents an unusual complication of anticoagulation and thrombolysis associated with the management of acute coronary syndrome. Conservative management was successful. PMID- 16352231 TI - Heart Foundation publications for health professionals. PMID- 16352230 TI - Patent ductus arteriosus: an uncommon cause of tracheobronchial compression in infants. AB - Tracheobronchial compression is an important cause of respiratory distress in children, requiring precise diagnosis and early surgical management. Common causes of the compression are either intrinsic or extrinsic, the latter being usually due to vascular rings. We report a 10 weeks old boy in whom a dilated pulmonary artery due a large patent ductus arteriosus was the cause for extrinsic compression. PMID- 16352232 TI - Victorian heart centre proceedings 2003. PMID- 16352233 TI - Valve sparing aortic root repairs--an anatomical approach. AB - While modern prostheses for aortic valve replacement provide a good substitute for the diseased aortic valve, they are still associated with a small but significant risk of complications. Thus, preservation of the native undiseased aortic leaflets during aortic root replacement is particularly attractive, especially if the repair is durable. Thorough understanding of the anatomy of the aortic valve and adjacent structures as well as the pathogenesis of aortic regurgitation in diseases of the aortic root is mandatory to the successful implementation of valve sparing aortic root repairs. PMID- 16352234 TI - Management of the elderly cardiac surgical patient: neurological sequelae. AB - The main neurological complications after cardiac surgery and non-surgical cardiac procedures include stroke, encephalopathy and neuropsychological deficits. Embolism from aortic atherosclerosis is the most common pathogenic mechanism. This article reviews the incidence, risk factors, and mechanisms of these complications. PMID- 16352235 TI - What's happening in paediatric cardiology? AB - Recent decades have witnessed dramatic advances in the care of adults with heart disease. However, equally significant advances have occurred in the care of children. In this review we describe some of the advances, which have been made in the care of children with heart disease, focusing not only on technological advances, but also on developments in team-based care, which together have resulted in significant improvements in outcomes. PMID- 16352236 TI - Robotic cardiac surgery at Epworth hospital. AB - Epworth Hospital is the first in the southern hemisphere to acquire the da Vinci Robot to facilitate minimally invasive cardiac surgery. Applications for this new technology include mitral valve repair, atrial septal defect closure, single coronary artery bypass graft to coronary arteries on the front of the heart, ablative surgery for atrial fibrillation and insertion of epicardial pacemaker electrodes. A team of surgeons from the Epworth Hospital have trained at East Carolina University (ECU) Greenville, North Carolina, USA; including surgeons, anesthetists, perfusionist and nurses. Following this, during a week in March, a proctoring surgeon from the USA, Dr. Wiley Nifong, assisted the Epworth team with their initial seven operations, which all proceeded without incident. The initial operative times were longer than the standard surgery, although with additional experience, now totalling 24 patients in all, these times have shortened considerably. The experience to date is summarized as an Addendum to this manuscript. PMID- 16352237 TI - BNP--Considering a heartfelt message. AB - The natriuretic hormones have been discovered as mediators of the cardiac response to volume overload and mechanical dysfunction. Although there are some physiological differences between atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), it is BNP that has been found to be most useful as a clinical test. BNP is secreted along with an N terminal proBNP cleavage product (NT-proBNP). The similarities between these two tests are far greater than the differences. They are both probably useful in the diagnosis of high-risk patients (e.g. dyspnoeic) and may be useful in monitoring cardiac failure treatment. Although BNP levels in the normal range predict the risk of cardiac events and mortality, there is little evidence to promote their use as a screening test. In such an important condition as cardiac failure we should consider any messages that are likely to be of value, particularly those that are heartfelt. PMID- 16352238 TI - Prevention of renal failure. PMID- 16352239 TI - "The Tobacco Case" document management by professional service providers: the ramifications of McCabe v British American Tobacco Australia Limited. AB - The legal issues concerning Document Management in Victoria, Australia, have achieved a level of notoriety as a result of the case of McCabe v British American Tobacco Australia Limited. The case has ramifications for all professional service providers. A Document Management Policy is a handy tool to have but it must be administered in a responsible and proper way. The Victorian Government is considering legislative reforms to ensure that this occurs. PMID- 16352240 TI - Hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy is a primary disorder of the myocardium characterised by disproportionate hypertrophy of the ventricular wall. It is the most common genetic cardiac disease with an incidence of 1 in 500 and it is diagnosed most commonly using transthoracic echocardiography. This review article discusses: the diagnosis of hypertrophic cardiomyopathy; the differential diagnoses; the characteristic histological signs found at postmortem and/or myectomy and the clinical symptoms and signs. Current recommendations for myectomy of first degree relatives, based on the ACC/ESC guidelines, are discussed as well as general management and then specific management for various subgroups and symptomatic patients. PMID- 16352241 TI - Cardiac resynchronisation therapy. AB - Left ventricular dysfunction is often associated with myocardial conduction slowing which is usually seen clinically as left bundle branch block on the surface ECG. Left bundle branch block causes asynchronous contraction of the left ventricle with the ventricular septum contracting early and the lateral left ventricular wall contracting late. This leads to a reduction in cardiac output and myocardial contraction efficiency and systolic mitral regurgitation worsens. Patients with this combination of findings may benefit from the implantation of a pacing system that aims to normalise conduction and "resynchronise" the ventricles. This mode of cardiac stimulation is referred to as "biventricular" pacing and relies on the implantation of an additional pacing lead on the epicardial surface of the left ventricle. This is achieved by selectively cannulating the coronary sinus and passing a pacing lead via a posterolateral coronary sinus tributary to an appropriate location. This lead, as well as the two conventional right atrial and right ventricular leads, is then attached to a specialised pacemaker. The procedure may be challenging and usually takes 1-2h depending on the operator's experience. The QRS complex shortens as the lateral wall of the left ventricle becomes "pre-excited" and contracts in concert with the ventricular septum. Improved haemodynamics result immediately and favourable reverse left ventricular remodelling occurs over subsequent weeks. Recently, these physiologic advantages have been translated into real clinical gains for patients with biventricular pacemakers where improvements in exercise tolerance and quality of life and reduced hospitalisations for recurrent heart failure have been conclusively demonstrated. PMID- 16352242 TI - Clinical trials and graft patency data in coronary artery surgery--a 30-year perspective. AB - The treatment of coronary artery disease, initially by medical and surgical and subsequently using percutaneous interventions, has evolved rapidly over the last 50 years. Randomized controlled clinical trials and observational studies of large cohorts have been instrumental in the evolution of the current practice of coronary artery revascularisation. This paper traces some of the historic trials of medical versus surgical therapy, surgical versus percutaneous intervention and graft patency studies. Medical therapy and percutaneous intervention are now the primary therapeutic modalities. However, there was an early recurrence rate with percutaneous intervention which has decreased with the use of drug-eluting stents. Coronary artery bypass surgery remains a major option for treating advanced coronary artery disease. Furthermore, it remains the treatment of first choice in most patients with left main coronary artery disease and those with extensive three-vessel proximal coronary artery disease, especially when associated with poor left ventricular function. The choice of a coronary artery bypass conduit is of major importance in developing a single long-term strategy for the surgical treatment of coronary artery disease. Bilateral internal thoracic artery grafting is now accepted as the treatment of choice, although not universally practiced. The role of other arterial conduits is being defined. PMID- 16352243 TI - The challenges of reconstructive surgery of thoracic defects. PMID- 16352244 TI - Spinal cord stimulation significantly improves refractory angina pectoris-a local experience spinal cord stimulation in refractory angina. AB - BACKGROUND: Severe refractory angina pectoris can occur in end-stage coronary artery disease despite maximal medical and revascularization therapy. Spinal cord stimulation is an under-utilized but well-established modality for the treatment of intractable angina pain. AIM: To illustrate the practical, beneficial and effective use of spinal cord stimulation as a treatment option for refractory angina in a local context. METHOD: A case series clinical audit of 11 patients with refractory angina treated with spinal cord stimulation over a one-year period was carried out. Baseline, three-month, six-month and two-year functional assessment data and subjective patient reports on their symptoms were evaluated. RESULTS: Spinal cord stimulation improves six-minute walk distance, exercise duration, New York Heart Association functional class, Likert score, and number of angina free days per week for at least one year over two years of follow-up. CONCLUSION: Spinal cord stimulation is an effective medium-term treatment option for refractory angina pectoris with significant benefits to functional parameters and patient symptoms. Spinal cord stimulation is an under-utilized but well established modality for the treatment of intractable angina pain. We report a small case series clinical audit of patients who gained significant functional and symptomatic benefits from this treatment. PMID- 16352245 TI - Cor-triatriatum sinister presenting in the adult as mitral stenosis: an analysis of factors which may be relevant in late presentation. AB - BACKGROUND: Cor-triatriatum sinister is a rare congenital heart defect resulting from the division of the left atrium by a fibro-muscular membrane. It is usual for patients to present in infancy and early childhood, although some cases remain undetected until adult life. As a consequence of trans-membrane flow obstruction, the clinical features often mimic mitral stenosis. At present, the reasons for late presentation are poorly understood. METHODS: A complete review of all cases of cor-triatriatum sinister published in the English literature from 1966 to date as mitral stenosis was performed. Statistical analysis was carried out to determine associations between measurements reflecting the communicating membrane fenestration area, the presence of several clinical variables and patient age at initial presentation. RESULTS: Both pulmonary capillary wedge pressure and mean pressure gradient were significantly higher in younger adults. In addition, the incidence of atrial fibrillation and mitral regurgitation was found to rise with advancing age. CONCLUSIONS: Cor-triatriatum sinister remains an uncommon form of congenital heart disease although it is being diagnosed with increasing frequency in adults due to improvements in diagnostic imaging. This diagnosis should be considered in all patients presenting with signs or symptoms of mitral stenosis. PMID- 16352246 TI - Endothelial function related to vascular tone in cardiac surgery. AB - Vascular endothelium has multiple functions including regulating of vascular tone, preventing platelet aggregation, anti-proliferation, etc. An intact endothelial function is essential to the maintenance of an adequate vascular tone, to prevent platelet aggregation in the intimal surface of blood vessels, to prevent smooth muscle proliferation, and to prevent atherosclerosis. This review focuses on endothelial function related to the vascular tone in cardiac surgery. The review is composed by three sections. In the first section, normal endothelial function related to vascular tone is described. In the second section, coronary endothelial function related to cardiac arrest and cardioplegic exposure is reviewed. In the third section, the endothelial function in the coronary bypass grafts is summarised. It is particularly important to understand that coronary endothelial dysfunction may be one of the major causes of low perfusion of the myocardium after cardiac arrest or donor heart preservation. Further, endothelium plays a major role in the maintenance of vascular tone and in the long-term patency of CABG grafts. The characteristics of endothelium in arterial and venous grafts and the correlation to the long-term patency are now more understood. A number of methods have been suggested to protect endothelial function in either coronary circulation or in coronary artery bypass grafts during cardiac surgery but further investigations in this field are warranted. PMID- 16352247 TI - Factors influencing the mean postoperative gradients across stentless porcine valves. AB - BACKGROUND: To study the preoperative and intraoperative variables influencing the mean post-operative transvalvular gradient across stentless porcine valves. METHODS: From 1995 to 2002, 84 patients underwent stentless valve insertion. The mean age was 73 years, and 63% were male. The valve pathology was aortic stenosis (AS) in 79%, aortic regurgitation (AR) in 12%, and mixed in 9%. Valve sizes ranged from 21 to 29 with size 27 being most frequent. 54% of patients had concomitant procedures. Patients had at least yearly clinical and echocardiographic follow-up. RESULTS: There was no operative mortality. 9.5% of the patients had significant postoperative complications. The average echo interval was 18.6 months (range 1-88). The overall mean transvalvular gradient was 9.88+/-5.67 (SD) mmHg. Variables associated with significantly reduced gradients were: larger valve sizes (p=0.002), younger age (p=0.05), pre-op AR (p=0.008), and increasing post-operative interval (p=0.05). The mean gradients decreased by 0.28 mmHg for each post-operative year. The method of implantation did not significantly affect gradients (p=0.26). CONCLUSIONS: Excellent mean transvalvular gradients were achieved with stentless valves studied, with a low operative risk. The gradients did not appear to be related to intra-operative factors, suggesting that insertion techniques can be tailored to suit patient conditions and surgeon preferences. PMID- 16352249 TI - The split radial technique: an alternative to the Y-graft for total arterial myocardial revascularisation. AB - There is evidence that the use of Y-grafts attached to the left internal mammary artery - to - left anterior descending artery graft may compromise the patency of the distal limb of the left internal mammary artery. We describe a technique (split radial technique) that avoids the use of Y-grafts by constructing two aorto-coronary grafts from a single radial artery. The split radial technique requires the harvesting of the radial artery in its entirety from the brachial bifurcation to the radial styloid. The first radial artery distal anastomosis is performed and the required length of conduit is determined. The conduit is transected, leaving a sufficiently long radial segment for a second aorto coronary graft. A clinical follow-up 41 weeks after surgery of the first 37 patients in whom the split radial technique was used showed no deaths or major complications. This suggests that the split radial technique is a useful and safe way to maximise the use of radial artery conduit and to avoid the potential risk of compromising internal mammary artery patency with Y-grafts. There is evidence that the use of Y-grafts attached to the left internal mammary artery may compromise the patency of the distal limb of the left internal mammary artery. We describe the split radial technique of constructing two aorto-coronary graft segments from a single radial artery that can be used to avoid the use of Y grafts. PMID- 16352248 TI - Ethyl pyruvate enhances ATP levels, reduces oxidative stress and preserves cardiac function in a rat model of off-pump coronary bypass. AB - BACKGROUND: Off-pump coronary artery bypass grafting is associated with transient periods of myocardial ischemia during revascularization resulting in myocardial contractile dysfunction and oxidative injury. The purpose of this study was to investigate the efficacy of ethyl pyruvate as a myocardial protective agent in a rat model of off-pump coronary artery bypass grafting associated with transient myocardial dysfunction without infarction. METHODS: Wistar rats were subjected to transient ischemia via 10 min occlusion of the LAD coronary artery followed by 10 min of reperfusion. Animals received an IV bolus of Ringer's solution as a control (n=10) or Ringer's ethyl pyruvate (n=10) immediately before the initiation of ischemia and reperfusion. Myocardial ATP and lipid peroxidation levels were quantified for an estimation of energetics and oxidative stress, respectively. In vivo cardiac function was assessed throughout the ischemia and reperfusion periods. RESULTS: Ethyl pyruvate significantly increased myocardial ATP levels compared to controls (2650+/-759 nmol/g versus 892+/-276 nmol/g, p=0.04). Myocardial oxidative stress was significantly reduced in animals treated with ethyl pyruvate compared to controls (70.4+/-2.6 nmol/g versus 81.8+/-2.4 nmol/g, p=0.04). dP/dt max and cardiac output were significantly greater in the ethyl pyruvate group compared to controls during ischemia and reperfusion. CONCLUSIONS: Ethyl pyruvate enhances myocardial ATP levels, reduces oxidative stress, and preserves myocardial function in a model of transient ischemia/reperfusion injury not subject to myocardial infarction. PMID- 16352250 TI - Ischemic preconditioning at a distance: altered gene expression in mouse heart and other organs following brief occlusion of the mesenteric artery. AB - Remote ischemic preconditionining (IPC) has been defined as a brief episode of ischemia/reperfusion in an organ that protects another remote organ from the damage induced by subsequent and prolonged ischemia. As yet, no study has been conducted with the purpose of elucidating a precise association between remote IPC and patterns of gene-transcription in cardiac tissue. In this study, using a cDNA microarray, we analyzed the gene expression profile in murine heart at 24h after brief cycles of occlusion of the superior mesenteric artery. The profile revealed that IPC induces significant levels of expression of many genes known to be associated with the stress response, redox regulation, growth and metabolism, DNA repair and other functions. The result of cDNA microarray profile from heart was also compared with those from other organs (lung, kidney and intestine). The genes identified in the expression profile may be associated with remote IPC induced late phase organ protection. PMID- 16352251 TI - Massive immature mediastinal teratoma extending into the left pleural cavity. AB - We report a case of a massive mediastinal teratoma in an 18-year-old woman who presented with a short history of exertional dyspnoea. The tumor arose from the left lobe of the thymus and extended into the left pleural cavity, completely compressing the left lung and extensively shifting the mediastinum to the right. Measuring 23 cm x 17 cm x 9 cm and weighing 2005 g it is one of the largest anterior mediastinal teratomas reported. It was successfully treated by surgical resection, with a final pathological diagnosis of a grades 1-2 immature teratoma. PMID- 16352252 TI - Primary lung cancer involving mediastinum and mimicking left atrial myxoma: a case report and review of literature. AB - We report a case of primary large cell carcinoma of the lung invading the heart and mimicking a left atrial myxoma. This case highlights the problems we face in diagnosis and management of a very unusual presentation of poorly differentiated primary large ('non-small') cell lung cancer with extension into the left atrium. It is important to have a high index of suspicion for a primary lung malignancy in chronic heavy smokers with none to minimal respiratory symptoms for early diagnosis and full staging procedures, as some of these tumours may be suitable for lung resection. PMID- 16352253 TI - Polycythaemia vera presenting as ST-elevation myocardial infarction. AB - A case of ST-elevation myocardial infarction as the first presentation of polycythaemia vera is described. The discussion summarises the evidence for the safety and efficacy of contemporary ST-elevation treatment strategies in the setting of polycythaemia vera. PMID- 16352254 TI - Operation for adult patent ductus arteriosus using an aortic stent-graft technique. AB - Patent ductus arteriosus is a condition usually diagnosed and treated in childhood but in some instances is not detected until adult life. Although surgical closure remains the accepted treatment in most cases, stent-grafting has emerged as a possible alternative treatment for persistent ductus arteriosus in the adult. We describe a case of elective repair of a patent ductus arteriosus using an aortic stent-graft technique in a young man. Successful use of the endoprosthetic approach may allow more widespread use of this technique in adult patent ductus arteriosus. PMID- 16352255 TI - Heart Foundation research beats stronger than ever before with help from our industry partners! PMID- 16352256 TI - Gender and age differences in the prevalence of coronary artery calcification in 953 Chinese subjects. AB - BACKGROUND: To investigate gender and age differences in coronary artery calcium (CAC) as determined by electron beam computed tomography (EBCT) in a Chinese population. METHODS: Consecutive patients undergoing EBCT were subdivided into groups based on gender and decades of life. They were further subdivided into three groups with respect to symptoms of coronary artery disease: typical, atypical and asymptomatic. Total calcium score was calculated for each patient and means calculated for each subgroup. Groups were then compared with respect to age, gender and symptoms. RESULTS: During the study period, 953 patients (736 men and 217 female) aged 17-86 years (mean 55+/-11 years) underwent EBCT. The prevalence of CAC increased significantly with increasing age. The mean total calcium score also increased with increasing age in males and females of each symptom subgroup. The prevalence of coronary artery calcification was significantly higher in males than females until age in excess of 69 years (p<0.05). The prevalence of coronary artery calcification and mean calcium scores were significantly different between each symptom subgroup (p<0.001) with higher scores and prevalence in patients with typical symptoms of coronary disease. CONCLUSIONS: There is an increase in the prevalence of coronary artery calcification with age in Chinese subjects. Male subjects are more likely than female subjects to have detectable coronary calcification up until an age in excess of 69 years. Patients under the age of 70 years, with typical symptoms of coronary artery disease have a higher prevalence and mean calcium score than those with atypical or no symptoms. PMID- 16352257 TI - Cost-benefit of drug eluting stents--time for a reality check. AB - BACKGROUND: Drug-eluting stents (DES) are currently being used in preference to bare stents in the public hospital system in 'high risk' lesions on the basis of cost-effectiveness analyses using data from trials which exaggerate their impact and do not include several categories of 'high risk'. This study aimed to assess the real cost-effectiveness of DES using prospectively collected outcome data from consecutive patients in the immediate pre-DES era. METHODS AND RESULTS: A prospective database on all percutaneous coronary interventions (PCI) performed by one operator in the two years prior to DES introduction (July 2000-June 2002) was used in which 12-month follow-up was complete in 96%. In 490 patients, 510 PCI were undertaken on 650 lesions (298 'high risk') using 686 stents. Angiography-proven clinical restenosis occurred in 31 lesions (5%) in 27 patients (12 focal, 19 diffuse; 11 treated with CABG, 14 with repeat PCI [4 re-restenosed 3 re-rePCI, 1 CABG] and 2 treated medically) with 2 deaths (both post-CABG). If all 'high risk' lesions had been treated with DES (assuming 0% clinical restenosis) then 22 lesions in 18 patients would not have restenosed avoiding 8 CABG and 13 re-PCI. Assuming A 5186 dollars cost for re PCI and A 18,496 dollars for CABG, DES will only be cost neutral for 'high risk' lesions when the premium for DES reduces to 617 dollars (currently 1500 dollars) and for all lesions at 452 dollars. CONCLUSIONS: Current strategies for use of DES are not cost effective. DES should be reserved for restenosis until the price comes down. PMID- 16352258 TI - Utility of brain natriuretic peptide (BNP) measurement in cardiovascular disease. AB - Cardiac failure is a prevalent and costly condition in Western society. The ageing of the population, together with current medical options which improve, rather than eradicate heart failure, lead to the projection that this problem will increase substantially in the foreseeable future. The availability of a simple test to assist the diagnosis and effective management of heart failure would greatly assist the clinical approach to this problem. This review examines the physiological basis for the measurement of natriuretic peptides as markers of the presence or risk of heart failure. It considers its use in the hospital and non-hospital setting and examines the cost-effectiveness of current assays. It is possible that in future natriuretic peptides may offer a form of treatment for heart failure, but this is beyond the scope of this review. Nevertheless, the review highlights the potential benefits of this group of tests in the management of heart failure. PMID- 16352259 TI - Post-infectious pseudoaneurysm after coronary angioplasty using drug eluting stents. AB - Infections at the site of coronary stents are uncommon, and believed to be the result of either direct stent contamination at the time of delivery, or from transient bacteraemia from the access site. A case of pseudoaneurysm ("mycotic" aneurysm) of the left anterior descending coronary artery, due to infection with Staphylococcus aureus after paclitaxel-eluting stents implantation is presented. PMID- 16352260 TI - A unique presentation of antiphospholipid antibody syndrome in pregnancy. AB - Primary antiphospholipid antibody syndrome (APS) is a protean disease with many manifestations including venous and arterial thrombosis, recurrent foetal loss, preeclampsia, intrauterine growth retardation, cardiac valvular disease, glomerulonephritis, thrombocytopaenia and livedo reticularis. We report an interesting case of a 19-year-old woman where the diagnosis of primary APS was initially made in the peripartum period. PMID- 16352261 TI - VATS lung biopsy in suspected, diffuse interstitial lung disease provides diagnosis, and alters management strategies. AB - OBJECTIVES: In patients with suspected diffuse interstitial lung disease, open lung biopsy is associated with high mortality (16%). This risk is only acceptable if diagnosis is made and management enhanced. We reviewed the role of VATS techniques in this group to determine the morbidity, mortality and outcomes in terms of diagnosis and enhanced management. METHODS: Over the period of 5 years, 78 patients with suspected diagnosis of diffuse interstitial lung disease on clinical and radiological grounds were referred to a single surgical team. The patients' case notes and histology reports were reviewed retrospectively. Correlation was made with histopathological diagnosis. RESULTS: All 78 patients had sufficient provision of material for histological analysis. Eight patients had a histological diagnosis not consistent with diffuse interstitial lung disease; in all eight patients, this significantly altered the subsequent management. Of the 70 patients with diffuse lung disease, 26 patients (37.1%) had a histological diagnosis of usual interstitial pneumonia. Thirteen patients (18.6%) had a histological diagnosis of unclassifiable diffuse lung disease despite an adequate biopsy. The remaining 31 patients (44.3%) had other positive histological diagnosis made. A difference between pre-operative clinico radiological and final histological diagnosis sufficient to change prognosis and definitive management was made in 19 patients (27.1%). The mean and median post operative stay was 2.8 days and 2 days, respectively. The in-hospital mortality was one patient (1.5%) due to adult respiratory distress syndrome. CONCLUSIONS: VATS lung biopsy can be performed in this group of patient with low mortality of 1.5%. It provides sufficient material for histological diagnosis in 100% of patients and alters the management and prognosis in a significant number of patients. We propose that the role of VATS and clinico-radiological techniques should be compared in a prospective controlled clinical trial. PMID- 16352262 TI - Video-assisted thoracoscopic (VATS) pleurodesis for malignant effusion: an Australian teaching hospital's experience. AB - BACKGROUND: Video-assisted thoracoscopic surgery (VATS) is commonly employed to achieve pleurodesis in patients suffering malignant pleural effusion (MPE). AIMS.: To evaluate the utility and outcome of VATS pleurodesis in management of MPE. METHODS: Two hundred and two consecutive VATS pleurodesis for MPE were evaluated. Data was derived from a prospectively maintained database and hospital records. Pleurodesis was deemed unsuccessful if a significant effusion occurred within 30 days of surgery. RESULTS: VATS pleurodesis was successful in 88% of patients (failure 12%) while recurrence of effusion occurred in 18%. Post operative air space, air leak, empyema and prolonged intercostal catheter drainage (>14 days) were all significantly associated with a failed procedure. Mean length of stay was 10.4 days and 42% of patients were discharged within 7 days of surgery. Morbidity was 20% with no operative deaths and median survival was 94 days. Inpatient mortality was 5%. High ASA (>or=4) was significantly associated with increased risk of inpatient death (p<0.001) and poorer long-term survival (43 days versus 133 days, p=0.05). CONCLUSIONS: VATS pleurodesis offers reasonable palliation of MPE with low morbidity and rapid recovery. Patients with an ASA score of >or=4 have a poor overall outcome and warrant less invasive palliative measures. PMID- 16352263 TI - Intrapericardial IGF-I improves cardiac function in an ovine model of chronic heart failure. AB - BACKGROUND: Following myocardial infarction, progressive deterioration of left ventricular function often follows, leading eventually to overt heart failure. In the myocardium, there is increased expression of insulin-like growth factor I (IGF-I) mRNA, protein and receptor levels, particularly in the peri-infarct zone, suggesting that IGF-I has a role to play in post-infarct cardiac structure and function. In this study, we examine the effects of exogenous IGF-I on cardiac function. METHODS: Intrapericardial IGF-I (15 microg/kg/d, n=3) or vehicle (sterile saline, n=3) was administered to sheep in chronic heart failure and the results of intrapericardial delivery compared with those of subcutaneous delivery. Left ventricular ejection fraction (EF) was measured to assess cardiac performance. Concentrations of plasma IGF-I were quantified by radioimmunoassay. RESULTS: Intrapericardial delivery of IGF-I resulted in a rapid and sustained increase (P<0.001) in EF, which remained elevated 14 days after cessation of treatment. Subcutaneous IGF-I treatment did not affect EF. Both subcutaneous and intrapericardial IGF-I administration increased concentrations of plasma IGF-I, although concentrations declined prior to the cessation of treatment. CONCLUSIONS: We conclude that the higher concentration of IGF-I in the myocardium, which results from intrapericardial delivery significantly increases EF in chronic heart failure but that subcutaneous delivery of IGF-I does not. PMID- 16352264 TI - Age-associated increase in creatine kinase-MB-specific activity in human serum confounds its use as an injury marker. AB - This study retrospectively examined a consecutive series of patient CKMB samples for a relationship between age (242 subjects, aged from 20 to 96 years old), serum CKMB activity and corresponding mass results analyzed for CKMB-specific activity (activity per molecule, unit/mg). Patients were categorized into six age groups. The means for specific activity of CKMB, standard error and 95% confidence intervals of each group were calculated. CKMB-specific activity in human serum increases with age regardless of gender or MI severity. As this greater propensity for increased CKMB activity may be due to alteration in part due to protein modification of post-MI release in the blood circulation, these findings add further weight for the current move to the use of troponins and other markers of myocardial injury. PMID- 16352265 TI - The aging myocardium: roles of mitochondrial damage and lysosomal degradation. AB - Myocardial aging, leading to circulatory dysfunction, complicates numerous pathologies and is an important contributor to overall mortality at old age. In cardiac myocytes, mitochondria and lysosomes suffer remarkable age-related alterations. Mitochondrial changes include structural disorganization and enlargement, while lysosomes, which are responsible for autophagic turnover of mitochondria, accumulate lipofuscin (age pigment), a polymeric, autofluorescent, undegradable material. These changes are caused by continuous physiological oxidative stress, and they advance with age because the cellular turnover machinery is inherently imperfect. Several mechanisms contribute to age-related accumulation of damaged mitochondria following initial oxidative injury. Such mechanisms may include clonal expansion of defective mitochondria, decreased propensity of altered mitochondria to become autophagocytosed (due to mitochondrial enlargement or decreased membrane damage associated with weakened respiration), suppressed autophagy because of heavy lipofuscin loading of lysosomes, and decreased efficiency of Lon and AAA proteases. Because lipofuscin laden lysosomes still receive newly synthesized lysosomal enzymes, even though they fail to degrade the pigment, the cells become in short supply of lysosomal hydrolases for functional autophagy, further limiting mitochondrial turnover. This interrelated mitochondrial and lysosomal damage eventually results in functional failure and death of cardiac myocytes. PMID- 16352266 TI - Coronary artery bypass surgery in undiagnosed traumatic pericardial rupture--25 years later. AB - A 55-year-old man was electively admitted for coronary artery bypass surgery. His admission chest X-ray showed an abnormal cardiac silhouette with complete leftward displacement. He had a past history of blunt thoracic trauma due to a motor vehicle accident treated conservatively. We present our findings and surgical difficulty during an operation on a patient with a previously undiagnosed pericardial rupture. This is the first reported case of its kind in the cardiac surgical literature. PMID- 16352267 TI - Multiple mediastinal metastasis of ovarian carcinoma. AB - Secondary mediastinal tumours are rare. We present a case report of multiple mediastinal metastasis of ovarian cancer, a very rare occurrence with only two cases previously reported in the literature. PMID- 16352268 TI - Intracardiac extension of metastatic pulmonary leiomyosarcoma. AB - Leiomyosarcomas are uncommon malignancies of soft tissue associated with a poor prognosis due to local and metastatic disease. We describe an example of metastatic pulmonary leiomyosarcoma with intracardiac extension in a patient who had previously undergone resection of a primary retroperitoneal mass. Distinguishing metastatic malignancy within the heart from a primary tumour, such as atrial myxoma, may be difficult, with the use of various imaging modalities assisting in diagnosis and management. In this case, the patient's course was complicated by dramatic progression with widespread emboli resulting in peripheral arterial occlusion and stroke. PMID- 16352269 TI - New frontiers for the journal. PMID- 16352270 TI - What becomes of the broken hearted? Coronary heart disease as a paradigm of cardiovascular disease and poor health among indigenous australians. PMID- 16352271 TI - Mechanical circulatory support for the failing heart: past, present and future. PMID- 16352272 TI - Preventable chronic diseases among indigenous australians: the need for a comprehensive national approach. AB - Chronic conditions such as diabetes, cardiovascular disease and heart disease represent a serious and escalating health burden for Indigenous populations across Australia. Social disadvantage, inactive lifestyle and poor nutrition are major contributors to chronic disease among Indigenous peoples. We have the knowledge base to reduce the devastating impact of early onset chronic diseases in those who already have the conditions, and to prevent or delay their onset in those who have not yet succumbed. This represents a major and urgent challenge to governments and Indigenous communities. PMID- 16352274 TI - Extra-ascending aortic versus intra-descending aortic balloon counterpulsation effect on coronary artery blood flow. AB - BACKGROUND: Diastolic counterpulsation has been used to provide circulatory augmentation for chronic heart failure or for short-term cardiac support. Recently an extra-aortic balloon (EAB) counterpulsation device has been proposed. AIM: To compare the circulatory effects of counterpulsation using the EAB or an intra-aortic balloon (IAB) in the acute pig model. METHODS: In six anaesthetized great white pigs (paced at 100 bpm), ECG, arterial and central venous pressures, flow in the coronary circulation and descending thoracic aorta were measured. Baseline data was collected, then with the EAB or an IAB fitted using 1:1 and 1:2 counterpulsation modes. Baseline data was compared to EAB and IAB data in 1:1 mode. Assisted beat data compared to unassisted beat data was also analysed in 1:2 mode. RESULTS: Both devices augmented peak diastolic arterial pressure, and decreased afterload. EAB counterpulsation increased diastolic coronary flow in both the 1:1 mode by 69% (p < 0.05) and in the 1:2 mode by 63% (assisted versus unassisted beat, p < 0.05). The IAB significantly increased diastolic coronary flow only in the 1:2 mode by 28% (p < 0.01). Both devices augmented total coronary flow and some augmentation of aortic flow was observed. CONCLUSION: The circulatory effect of the EAB and IAB counterpulsation were comparable. This suggests the EAB could be used as a non-blood contacting heart assist device in patients suffering moderate-severe heart failure. PMID- 16352273 TI - Adverse reactions of low osmolar non-ionic and ionic contrast media when used together or separately during percutaneous coronary intervention. AB - BACKGROUND: Due to perceived advantages in the use of non-ionic contrast agents for diagnostic angiography and ionic agents for percutaneous coronary intervention (PCI), patients often receive various combinations of both types of agents. AIM: To assess potential adverse effects of non-ionic and ionic contrast media when used together or separately during percutaneous coronary intervention. METHODS: We retrospectively evaluated the outcomes of 532 patients undergoing percutaneous coronary intervention in our institution. Patients were divided into two groups: those that underwent diagnostic angiography and "follow on" PCI; and those that underwent "planned" PCI. The groups were subdivided on the basis of the use of the ionic agent ioxaglate or the non-ionic agent iopromide during PCI. The frequency of allergic reactions and major adverse cardiac events (MACE) were noted. RESULTS: With respect to the "follow on" group, allergic reactions occurred in 9 of 150 patients (6.0%) who received the combination of ioxaglate and iopromide versus 1 of 93 (1.1%) who only received iopromide (p=0.094). There was no difference with respect to MACE [6 (4.0%) ioxaglate and iopromide versus 4 (4.3%) iopromide alone, p=1.00]. In the "planned" group, 7 of 165 patients (4.2%) receiving ioxaglate had an allergic reaction as opposed 0.0% (0 of 124 patients) in the iopromide group (p=0.021). All contrast reactions were mild. The incidence of a MACE was similar in both groups [1 (0.6%) ioxaglate versus 2 (1.6%) iopromide, p=0.579]. The incidence of allergic reactions was similar if ioxaglate was used alone or in combination with iopromide (p=0.478). CONCLUSIONS: Whilst combining ionic and non-ionic contrast agents in the same procedure was not associated with any more adverse reactions than using an ionic contrast agent alone, the ionic contrast agent ioxaglate was associated with the majority of allergic reactions. With respect to choice of contrast agent, using the non-ionic agent iopromide alone for coronary intervention is associated with the lowest risk of an adverse event. PMID- 16352275 TI - The outcome of cardiac surgery in dialysis-dependent patients. AB - BACKGROUND: Patients on dialysis for end-stage renal failure (ESRF) are undergoing cardiac surgery with increasing frequency. Furthermore, ESRF is known to be an important risk factor for complications of cardiac operations performed with cardiopulmonary bypass. AIMS: To evaluate the outcome of dialysis-dependent patients undergoing cardiac surgery at one institution. METHODS: A retrospective analysis was performed on consecutive patients with ESRF dependent upon maintenance haemodialysis or peritoneal dialysis who underwent cardiac surgery from January 1998 to August 2002. RESULTS: Thirty-eight patients on dialysis underwent cardiac surgery during this time period (1.5% of total cases). The most common cause for ESRF was diabetic nephropathy (n = 12). Operations performed included isolated coronary artery bypass grafting (CABG, n = 22), CABG and valve surgery (n = 8), and valve surgery alone (n = 6). When allowing for age, sex, surgeon and operative category, the odds ratio for mortality risk of dialysis patients, compared with all others, was 4.9 (95% confidence interval (CI): 1.7 13.9, p = 0.003), and for morbidity risk, was 2.8 (95% CI: 1.4-5.4, p = 0.003). CONCLUSIONS: Patients on dialysis have an increased morbidity and mortality following cardiac surgery, however we believe ESRF should not be regarded as an absolute contraindication to cardiac surgery or cardiopulmonary bypass. PMID- 16352276 TI - Return to work after coronary artery bypass surgery in a population of long-term survivors. AB - BACKGROUND: Return to paid employment may be facilitated by coronary artery bypass graft (CABG) surgery. We assessed work status in a population-based study of long-term outcomes of CABG. AIM: To determine the association between returning to work after CABG and clinical and socio-demographic factors. METHODS: A postal survey of 2,500 randomly selected patients 6-20 years post-CABG. The outcomes assessed were work status in the year before and after CABG and health related quality of life (HRQOL) measured with SF-36. RESULTS: Response was 82% (n = 2,061). Employment fell from 56% in the year prior to CABG to 42% in the year after. Workers in 'blue-collar' occupations were more likely to reduce their work status than those in 'white collar' occupations (46% versus 29%, p < 0.001). Independent predictors of reducing employment were increasing age (9% per year, 99% CI: 1.06-1.11, p < 0.001), 'blue-collar' versus 'white collar' occupation (OR: 2.1, 99% CI: 1.4-3.1) and female sex (OR: 2.1, 99% CI: 1.1-3.6). HRQOL among participants under 60 years of age at follow-up was better for those who returned to work after CABG surgery. CONCLUSION: CABG surgery is followed by a net loss to paid employment of working age patients which increases with age, and is more likely for those in blue-collar occupations and women. PMID- 16352277 TI - Intravascular ultrasound-guided percutaneous coronary intervention in a human immunodeficiency virus-positive patient with cocaine-associated acute myocardial infarction: case report and review. AB - Cocaine use is a major problem worldwide and there are numerous reports about cocaine-associated myocardial infarction. Nevertheless minimal data are available from randomised clinical trials to suggest evidence-based approaches to the management of cocaine-associated myocardial ischemia. Moreover, most reports have been limited to conservative management of cocaine-associated myocardial infarction. We report a case of a young male cocaine user with acute myocardial infarction, undergoing diagnostic coronary angiography and intravascular ultrasound revealing severe atherosclerosis, followed by successful stent implantation. PMID- 16352278 TI - Proceedings of the Victorian Heart Centre. PMID- 16352279 TI - Arterial conduits update. AB - Arterial grafting for coronary artery disease has been practiced routinely at the Epworth Hospital for 20 years. Bilateral versus single internal thoracic artery grafts is associated with improved survival, which progressively increased over the 20 years of post-surgery. The average increase in survival was 15% at 20 years. The question of whether to use the radial artery or the saphenous vein graft as supplementary grafts remains unanswered. Current patency data suggest that there is a marginal advantage of radial artery over saphenous graft patency, particularly between 5 and 10 years after surgery. However, there appears to be improved patency of the saphenous vein when used as a coronary artery bypass with the advent of aspirin, lipid-lowering and anti-hypertensive therapy. PMID- 16352280 TI - Cardiac involvement in Fabry's disease. AB - Clinical Fabry's disease is due to any of multiple mutations in the X-linked alpha-galactosidase gene. These mutations are kindred-specific, often spontaneous, and produce varying degrees of functional enzyme deficiency resulting in deposits of specific glycosphingolipid (cerumide), especially in the vasculature, kidneys, heart and reticuloendothelial tissue. Disease frequency has probably been over-estimated at 1/40,000; so few centres have developed clinical experience of the disease, though the disease has been identified in all major racial groups. PMID- 16352281 TI - Obstructive sleep apnoea, congestive heart failure and cardiovascular disease. AB - Sleep disordered breathing is a common condition within the general community. Mostly this is represented by obstructive sleep apnoea (OSA), a condition characterized by repetitive occlusions of the upper airway due to retro positioning of the tongue and pharyngeal collapse during sleep. This article covers the key evidence relating OSA to both causation and progression of congestive heart failure and cardiovascular disease including hypertension. The results of recent studies are summarized, and the authors conclude that whilst progress has been made, there remain many gaps in our knowledge in relation to the contribution to the burden of cardiac disease produced by associated conditions such as OSA. Larger studies with important primary endpoints will be required to demonstrate the merit of screening and treating this disorder. PMID- 16352282 TI - Radiofrequency ablation for the treatment of atrial fibrillation--brief report update. AB - Considerable advances have been made in both the understanding of and the curative treatment for atrial arrhythmias over the last decade. Radiofrequency ablation of typical right atrial flutter and focal atrial tachycardia using standard catheters is commonplace and is associated with success rates of over 95%. Recently developed 3-D mapping systems have furthered our understanding of the complex circuits responsible for atypical atrial flutters within the right and left atrium and successful ablation of these previously incurable arrhythmias is now routinely achieved. With the seminal discovery of pulmonary vein triggers and fixed left atrial circuits that are critical for the genesis and perpetuation of atrial fibrillation, ablation techniques have been developed which are associated with cure rates of over 75% for both paroxysmal and persistent forms of atrial fibrillation. PMID- 16352283 TI - Medicine in the Indian Himalaya--experiences of a volunteer doctor. PMID- 16352284 TI - Nuclear cardiology. AB - Nuclear cardiology has shown double digit growth yearly over the last few years and remains the second most commonly performed noninvasive cardiac imaging investigation of choice. Approximately 9 million studies performed yearly in the U.S. alone. As cardiovascular disease remains the largest healthcare problem around the world despite increasing healthcare spending there is growing interest to enhance the use of cost effective diagnostic tools, such as nuclear cardiology. Recent development of technology has seen the addition of hybrid imaging tools such PET/CT and SPECT/CT into the armorial. More data is expected to become available on this subject within the next few years, and we also expect several molecular imaging tests to become available for clinical use. Despite easy availability of these diagnostic tools, cardiac mortality and morbidity in patients with long term diabetes and also in the postoperative setting remains unexpectedly high and emergency department physician are still being sued after patients being discharged with undiagnosed chest pain. The value of the use of nuclear cardiology studies is briefly reviewed in these clinical settings. PMID- 16352285 TI - The Heartpod implantable heart failure therapy system. AB - Congestive heart failure has been described as the new epidemic, despite advances in drug therapy a 2-year mortality of up to 50% persists. This is despite recent improvements in drug therapy. There are limitations to the current drug treatments and cardiac resynchronization devices. The treatment of diastolic dysfunction can be suboptimal. The Savacor Company developed the Heartpod device to directly measure left atrial pressure in patients with congestive heart failure via an implantable device. The patient can in real time, download their intra-cardiac pressure measurements to a hand-held device. With this information, they can titrate their own treatment in a very precise manner. PMID- 16352286 TI - Epworth hospital electrophysiology and pacing report, 2004-2005. AB - Although pacing and electrophysiology (EP) have been practised at the Epworth since the mid-1980s, they have enjoyed significant and continued growth since the opening of the J.G. Sloman dedicated EP laboratory 4 years ago. Nevertheless, there has not been a formal audit performed in these fields so far. This analysis sought to define the number and nature of the cases treated in the EP and pacing laboratory over a period of 18 months, so that recommendations might be made to improve our future practise in this area. Whilst it became apparent that the lack of a uniform and consistent data base (such as that which is used for the interventional cardiology laboratory) significantly hampered the process of this analysis, nevertheless the analysis revealed the high standard of performance and level of success attained in these cases over the given time period. PMID- 16352287 TI - Cardiovascular magnetic resonance imaging. PMID- 16352288 TI - Evidence-based ICU feeding algorithm--the Epworth experience. PMID- 16352289 TI - Assessment of myocardial viability in ischemic cardiomyopathy. AB - The number of patients with ischemic cardiomyopathy has increased extensively over recent years. Therapies include medical treatment, cardiac transplantation, cardiac resynchronization therapy and surgery. In the diagnostic and prognostic work-up, the assessment of myocardial viability has become more important over time. In particular, patients with viable myocardium can improve in LV function after revascularization; this will not occur in patients without viable tissue. In view of the high risk of surgery in this patient category, careful analysis is needed in order to justify the enhanced risk. Over the years, different viability techniques have been developed. In this review, these techniques are discussed. Moreover, the value of these techniques for the prediction of not only improvement of function, but also improvement in exercise capacity, reverse LV remodeling, and long-term prognosis, is discussed. PMID- 16352290 TI - Chemiluminescence-based electrophoretic mobility shift assay of heparin-protein interactions. PMID- 16352291 TI - The role of cognitive factors in the pathogenesis of obsessive-compulsive symptoms: a prospective study. AB - Cognitive models of obsessive-compulsive disorder (OCD) posit that specific kinds of dysfunctional beliefs (e.g., pertaining to responsibility and the significance of intrusive thoughts) underlie the development of this disorder. The present study was designed to prospectively evaluate whether dysfunctional beliefs thought to underlie OCD act as a specific vulnerability factor in the pathogenesis of obsessive-compulsive symptomatology. Eighty-five individuals were prospectively followed over a period of time thought to be associated with an increased onset of OCD symptoms -- childbirth and the postpartum. The majority of these new mothers and fathers experienced intrusive infant-related thoughts and performed neutralizing behaviors similar to, but less severe than, those observed in OCD. Scores on a measure of dysfunctional beliefs thought to underlie OCD predicted the development of obsessive-compulsive symptoms after controlling for pre-existing OCD symptoms, anxiety, and depression. Dysfunctional beliefs also predicted the severity of checking, washing, and obsessional OCD symptom dimensions, but not neutralizing, ordering, or hoarding symptom dimensions. These data provide evidence for specific dysfunctional beliefs as risk factors in the development of some types of OCD symptoms. PMID- 16352292 TI - Identification and characterization of the DNA-binding properties of a Zhangfei homologue in Japanese pufferfish, Takifugu rubripes. AB - Zhangfei is a basic region-leucine zipper (bZIP) transcription factor identified through its interaction with a herpesvirus-related host cell factor HCF1 (C1). Unlike most bZIP proteins, the mammalian Zhangfei protein does not bind DNA as homodimers. It is believed due to the absence of an asparagine residue in the basic region, which forms the DNA-recognition motif, NxxAAxxCR, in all bZIP proteins. Here, we report the identification and characterization of a novel Zhangfei homologue in Takifugu rubripes, which has an intact DNA-recognition motif by sequence analysis. We found that the pufferfish Zhangfei (pZF) appeared to have all the functional domains known in human Zhangfei, including the conserved HCF1-binding motif; however, pZF did not appear to bind DNA either. These findings suggest that the distinct property of the Zhangfei basic region is conserved during the evolution of vertebrates and that Zhangfei requires interaction with other proteins to regulate transcription from target promoters. PMID- 16352293 TI - Localization of GABA (gamma-aminobutyric acid) markers in the turtle's basal optic nucleus. AB - Recent physiological data have demonstrated that retinal slip, the sensory code of global visual pattern motion, results from complex interactions of excitatory and inhibitory visual inputs to neurons in the turtle's accessory optic system (the basal optic nucleus, BON). In the present study, the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), its synthetic enzyme, glutamic acid decarboxylase (GAD-67) and its receptor subtypes GABA(A) and GABA(B) receptors were localized within the BON. GABA antibodies revealed cell bodies and processes, whereas antibodies against GAD revealed a moderate density of immunoreactive puncta throughout the BON. GAD in situ hybridization labeled BON cell bodies, indicating a possible source of inhibition intrinsic to the nucleus. Ultrastructural analysis revealed terminals positive for GAD that exhibit symmetric synaptic specializations, mainly at neuronal processes having small diameters. Neurons exhibiting immunoreactivity for GABA(A) receptors were diffusely labeled throughout the BON, with neuronal processes exhibiting more labeling than cell bodies. In contrast, GABA(B)-receptor-immunoreactive neurons exhibited strong labeling at the cell body and proximal neuronal processes. Both these receptor subtypes are functional, as evidenced by changes of visual responses of BON neurons during application to the brainstem of selective receptor agonists and antagonists. Therefore, GABA may be synthesized by BON neurons, released by terminals within its neuropil and stimulate both receptor subtypes, supporting its role in mediating visually evoked inhibition contributing to modulation of the retinal slip signals in the turtle accessory optic system. PMID- 16352297 TI - Evidence for multiple Src binding sites on the alpha1c L-type Ca2+ channel and their roles in activity regulation. AB - OBJECTIVE: Src has been proposed to activate L-type calcium channel activity by binding to the alpha1c subunit. In the II-III linker region of this subunit there is a novel consensus sequence for Src binding. We have examined whether this site is a functional Src interaction site and investigated the effect displacing Src from this region has on calcium channel activity. METHODS: In vitro binding assays were performed to map alpha1 subunit interaction sites. Cardiac myocytes were isolated enzymatically from rat ventricles. Whole cell patch-clamp technique was used to record Ca(2+) channel currents in cells that had been loaded with the Src inhibitor PP1 and/or peptides with amino acid sequence corresponding to the hypothesized Src docking site. Co-immunoprecipitation and pull-down studies were undertaken to identify proteins co-complexing with the alpha1 subunit. RESULTS: Peptides corresponding to the II-III linker region and C-terminal tail of the alpha1c subunit, but not scrambled peptide controls, were found to inhibit Src SH3 domain binding to the channel and significantly reduced the channel current amplitude. The II-III linker region peptide shifted the inactivation curve to the left whereas the C-terminal tail region peptide shifted the activation curve to the right when compared to scramble peptide controls. PP1-pre-treatment of myocytes also reduced the current amplitude, decreased the V(1/2) for channel inactivation and abolished any further effect on currents by Src binding peptides. The tyrosine kinase PYK2 was found to co-associate with Src and the channel, but PP1 pre-treatment reduced this co-association. CONCLUSIONS: Src binds to both the II-III linker and C-terminal tail regions of the alpha1c subunit to differentially modulate channel activity. PYK2 is also able to co complex with Src when bound to this region of the channel but only when Src is catalytically active. Together the two kinases may synergistically regulate channel activity. PMID- 16352298 TI - Vimang (Mangifera indica L. extract) induces permeability transition in isolated mitochondria, closely reproducing the effect of mangiferin, Vimang's main component. AB - Mitochondrial permeability transition (MPT) is a Ca(2+)-dependent, cyclosporin A (CsA)-sensitive, non-selective inner membrane permeabilization process. It is often associated with apoptotic cell death, and is induced by a wide range of agents or conditions, usually involving reactive oxygen species (ROS). In this study, we demonstrated that Mangifera indica L. extract (Vimang), in the presence of 20 microM Ca(2+), induces MPT in isolated rat liver mitochondria, assessed as CsA-sensitive mitochondrial swelling, closely reproducing the same effect of mangiferin, the main component of the extract, as well as MPT-linked processes like oxidation of membrane protein thiols, mitochondrial membrane potential dissipation and Ca(2+) release from organelles. The flavonoid catechin, the second main component of Vimang, also induces MPT, although to a lesser extent; the minor, but still representative Vimang extract components, gallic and benzoic acids, show respectively, low and high MPT inducing abilities. Nevertheless, following exposure to H(2)O(2)/horseradish peroxidase, the visible spectra of these compounds does not present the same changes previously reported for mangiferin. It is concluded that Vimang-induced MPT closely reproduces mangiferin effects, and proposed that this xanthone is the main agent responsible for the extract's MPT inducing ability, by the action on mitochondrial membrane protein thiols of products arising as a consequence of the mangiferin's antioxidant activity. While this effect would oppose the beneficial effect of Vimang's antioxidant activity, it could nevertheless benefit cells exposed to over production of ROS as occurring in cancer cells, in which triggering of MPT mediated apoptosis may represent an important defense mechanism to their host. PMID- 16352299 TI - Quantitative analysis of tiropramide in human plasma by gas chromatography coupled to mass spectrometry for application to a bioequivalence test. AB - BACKGROUND: Tiropramide is used as an antispasmodic agent. A sensitive, selective and simple gas chromatography coupled to mass spectrometry (GC/MS) was developed for quantification of tiropramide in human plasma using internal standard (ISD, (+/-) alpha-benzoylamino-4-[2-(dimethylamino) ethoxy]-N,N dipropylbenzenepropanamide). METHODS: Tiropramide and ISD were extracted from plasma by solid-liquid extraction and analyzed on a HP-5MS column with mass selective detector. RESULTS: The retention times of tiropramide and ISD were approximately 9.8 and 10.2 min, respectively. The calibration curve showed good linearity in the concentration range 5-500 ng/ml (r2=0.998) for tiropramide in human plasma and showed good precision with CVs between 0.24% and 7.69%, respectively. The method was showed good accuracy with all intra-day (n=5) and inter-day (n=5) mean concentrations within 87.9-114.1% from nominal. The recovery of tiropramide and ISD were about 75.1% and 71.0% on the average, respectively. This method was successfully applied for the bioequivalence test of 2 formulations of tiropramide in 18 healthy male volunteers who received a single 100 mg dose of each formulation. PMID- 16352300 TI - A multiresolution diffused expectation-maximization algorithm for medical image segmentation. AB - In this paper a new method for segmenting medical images is presented, the multiresolution diffused expectation-maximization (MDEM) algorithm. The algorithm operates within a multiscale framework, thus taking advantage of the fact that objects/regions to be segmented usually reside at different scales. At each scale segmentation is carried out via the expectation-maximization algorithm, coupled with anisotropic diffusion on classes, in order to account for the spatial dependencies among pixels. This new approach is validated via experiments on a variety of medical images and its performance is compared with more standard methods. PMID- 16352301 TI - Estimations of past male and female serum concentrations of biomarkers of persistent organochlorine pollutants and their impact on fecundability estimates. AB - Persistent organochlorine pollutants (POPs) have been suggested to have negative effects on a number of hormonal systems. Several studies performed retrospectively have reported a possible association between POP exposure and fertility, measured as time to pregnancy (TTP). However, these studies lack biomarkers of exposure at the time when the women tried to conceive. It has previously been found that past female serum concentrations of 2,2',4,4',5,5' hexachlorobiphenyl (CB-153) can be estimated using a complex decay model, assuming that the biological half-life is 5 years, the yearly environmental reduction of the compound has been 3% since 1976, and the reduction of body burden due to lactation is 20% for periods up to 6 months and 30% for periods exceeding 6 months. In the present study, it is established that the model is valid also for estimations of past male serum concentrations of CB-153. Furthermore, the complex decay model was found to be useful also for estimating past serum concentrations of 1,1-dichloro-2,2-bis (p-chlorophenyl)-ethylene (p,p' DDE), assuming that the biological half-life of the compound is 8 years, the yearly reduction between 1971 and 1981 was 20% and after that 9%, and the reduction of body burden due to lactation is the same as that for CB-153. However, even though the estimated past serum concentrations of CB-153 and p,p' DDE were found to be better proxy measures of actual past concentrations than current serum concentrations, there was little change in the rank order of the population investigated. Thus, the effect estimate for TTP was similar for both proxy measures when using categorized measures of exposure. PMID- 16352302 TI - Lead exposure and eclampsia in Britain, 1883-1934. AB - Eclampsia refers to a coma or seizure activity in a pregnant woman with no prior history of such activity. This paper presents a mix of historical and epidemiological evidence consistent with the hypothesis that chronic lead exposure is a predisposing factor for eclampsia. The historical evidence is based on research conducted by British physicians around 1900 showing that the geographic variation in eclampsia across England and Wales was correlated with lead levels in local drinking water supplies. A formal epidemiological analysis based on a data set of English and Welsh counties observed in 1883 corroborates the evidence presented by historical observers. In particular, the statistical results show that the death rate from eclampsia in counties with high-water-lead levels exceeded the death rate in counties with low-water-lead levels by a factor of 2.34 (95% CI: 1.54-3.14). PMID- 16352303 TI - Determination of key amino acids implicated in the actions of allosteric modulation by 3,3'-difluorobenzaldazine on rat mGlu5 receptors. AB - Several mutations in the seven-transmembrane region of rat metabotropic glutamate 5 (rmGlu5) receptors were produced by site-directed mutagenesis and expressed in CHO cells. Using functional intracellular calcium ([Ca(2+)](i)) mobilisation, we identified amino acids implicated in the positive allosteric modulation of quisqualate-induced response by 3,3'-difluorobenzaldazine (DFB). Human and rat mGlu5 receptors displayed a higher potency and a higher efficacy in the presence of DFB. Mutant receptors S657(3.39)C, T780(6.44)A and M801(7.39)T disrupted the DFB-mediated increase in functional response. DFB-induced increase in potency was abolished in mutant receptors N733(45.51)A, Y791(6.55)A, A809(7.47)V, P654(3.36)S/S657(3.39)C and P654(3.36)S/S657(3.39)C/L743(5.47)V without affecting the enhancement of efficacy observed in wild type receptors. Mutations at positions Leu-743(5.47) and Trp-784(6.48) resulted in significantly larger DFB induced potentiation of EC(50) and E(max) values than in wild type receptors. DFB mediated increase of efficacy was abolished and EC(50) values were right-shifted in mutant receptor F787A, resulting in DFB acting as a weak partial antagonist at this mutant receptor. Based on these findings, we constructed a homology model concluding that six key residues in transmembranes 3, 5, 6 and 7 are necessary for the allosteric modulation of rmGlu5a receptor by DFB. The model confirms an overlapping but distinct binding site to 2-methyl-6-(phenylethynyl)-pyridine (MPEP), and in particular emphasises the key role of W784 in transmembrane (TM) 6 for controlling the receptor's activation state. PMID- 16352304 TI - Differential activation of ERK, p38, and JNK MAPK by nociceptin/orphanin FQ in the potentiation of prostaglandin cerebrovasoconstriction after brain injury. AB - Fluid percussion brain injury elevates the cerebrospinal fluid (CSF) concentration of the opioid nociceptin/orphanin FQ (N/OFQ), which potentiates vasoconstriction to the prostaglandins U 46619, a thromboxane A(2) mimic, and prostaglandin (PG)F(2a). This study investigated the role of the extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK) isoforms of mitogen activated protein kinase (MAPK) in potentiated prostaglandin vasoconstriction after brain injury and the relationship of brain injury induced release of N/OFQ to MAPK. Pial artery diameter was measured with a video microscaler by observation through a glass coverslip cranial window placed in the parietal cortex of newborn pigs. Brain injury potentiated U 46619 induced pial artery vasoconstriction but U 0126 and SB 203580 (10(-6) and 10(-5) M, respectively) (ERK and p38 MAPK inhibitors) blocked the potentiation. In contrast, administration of SP 600125 (10(-6) and 10(-5) M) (JNK MAPK inhibitor) only attenuated brain injury induced U 46619 potentiation and such responses were significantly different than that in the presence of either U 0126 or SB 203580 after FPI. Co-administration of N/OFQ (10(-10) M), the CSF concentration observed after brain injury, with U 46619 or PGF(2a) under non brain injury conditions potentiated prostaglandin vasoconstriction but U 0126 and SB 203580 blocked such potentiation. Administration of SP 600125 modestly attenuated prostaglandin potentiation by N/OFQ. These data show that activation of ERK and p38 primarily contribute to potentiation of prostaglandin constriction after brain injury. These data suggest that N/OFQ differentially activates ERK, p38, and JNK MAPK to contribute to potentiated prostaglandin vasoconstriction after fluid percussion brain injury. PMID- 16352305 TI - Growth hormone activity in mitochondria depends on GH receptor Box 1 and involves caveolar pathway targeting. AB - Growth hormone (GH) binding to its receptor (GHR) initiates GH-dependent signal transduction and internalization pathways to generate the biological effects. The precise role and way of action of GH on mitochondrial function are not yet fully understood. We show here that GH can stimulate cellular oxygen consumption in CHO cells transfected with cDNA coding for the full-length GHR. By using different GHR cDNA constructs, we succeeded in determining the different parts of the GHR implicated in the mitochondrial response to GH. Polarography and two-photon excitation fluorescence microscopy analysis showed that the Box 1 of the GHR intracellular domain was required for an activation of the mitochondrial respiration in response to a GH exposure. However, confocal laser scanning microscopy demonstrated that cells lacking the GHR Box 1 could efficiently internalize the hormone. We demonstrated that internalization mediated either by clathrin-coated pits or by caveolae was able to regulate GH mitochondrial effect: these two pathways are both essential to obtain the GH stimulatory action on mitochondrial function. Moreover, electron microscopic and biochemical approaches allowed us to identify the caveolar pathway as essential for targeting GH and GHR to mitochondria. PMID- 16352306 TI - Risk factors for deep infection in secondary intramedullary nailing after external fixation for open tibial fractures. AB - BACKGROUND: Risk factors for deep infection in secondary intramedullary nailing (IMN) after external fixation (EF) for open tibial fractures were investigated by multivariate analysis following univariate analyses. METHODS: Forty-two open tibial fractures were treated with secondary IMN after EF. The open tibial fractures were classified according to the criteria proposed by Gustilo et al.: type II, 11; type IIIA, 8; type IIIB, 22 and type IIIC, 1. Locked IMNs with limited reaming were performed in 27 patients, and locked IMNs without reaming in 15 patients. The following factors contributing to deep infection were selected for analysis: age, gender, Gustilo type (II or III), fracture grade by AO type (A or B+C), fracture site, existence of multiple trauma (Injury Severity Score, ISS<18 or ISS> or = 18), existence of floating knee injury, debridement time (< or = 6 h or > 6 h), reamed (R) versus unreamed (UR) nailing, duration of external fixation (< or = 3 weeks or >3 weeks), interval between removal of EF and IMN (< or = 2 weeks or >2 weeks), skin closure time (< or = 1 week or >1 week), existence of superficial infection (+ or -) and existence of pin tract infection (+ or -). The relationship between deep infection and the above factors was evaluated by univariate analyses. RESULTS: Seven (16.7%) of the 42 open tibia fractures developed deep infections. All deep infections occurred in Gustilo type III (22.6%, 7/31). Only the skin closure time was a significant factor affecting the occurrence of deep infection on the present analysis (p = 0.006). CONCLUSION: The present evaluation showed that early skin closure within 1 week is the most important factor in preventing deep infections when treating open tibial fractures with secondary IMN after EF. PMID- 16352307 TI - Conflict of interest: legal and ethical aspects. AB - Conflicts of interest arise when those who owe conscientious duties to others appear to have personal interests that might tempt them to subordinate those duties to their self-interest. Conflicts of interest are distinguishable from conflicts of commitment, which arise when individuals bear two or more mutually exclusive duties to others. If persons in conflicts of interest favor their self interest, they may violate binding legal duties such as fiduciary duties. Conflicts arise when, for instance, medical practitioners refer their patients to other practitioners in exchange for payments (fee splitting), and when they gain secondary incomes such as from testing patients' samples in laboratories in which they hold ownership or investment interests. Financial dealings with commercial sponsors may place researchers in apparent conflicts of interest with research subjects. When conflicts of interest are unavoidable, they may be resolved by appropriate disclosure. PMID- 16352308 TI - C-reactive protein 3' UTR +1444C>T polymorphism in patients with spontaneous venous thromboembolism. AB - OBJECTIVE: Data on C-reactive protein (CRP) as a risk indicator of venous thromboembolism are conflicting. A recent study reported higher CRP levels in homozygous carriers of a novel CRP gene polymorphism at the 3' UTR (CRP +1444C>T). We investigated, whether homozygosity for CRP +1444C>T is associated with an increased risk of spontaneous venous thromboembolism (VTE). METHODS AND RESULTS: CRP +1444C>T genotype and plasma levels were assessed in 128 patients with deep venous thrombosis (DVT, 70 females/58 males), 105 with pulmonary embolism (PE, 58 females/47 males) and 122 healthy individuals (60 females/62 males). CRP +1444TT was significantly associated with increased CRP plasma levels in healthy individuals. CRP +1444TT was more frequent (14%) among controls than DVT patients (9%, p=0.26) or PE patients (6%, p=0.05), respectively. No significant deviation from Hardy-Weinberg equilibrium was observed in patients (p=0.8) or controls (p=0.3), respectively. CRP +1444C>T was not significantly associated with CRP levels in patients with VTE. CONCLUSIONS: Homozygous carriers of the CRP 3' UTR +1444C>T polymorphism do not have a significantly increased risk of VTE. Our data support the assumption that a clinically relevant association between CRP and VTE is missing. PMID- 16352309 TI - Development of quantitative structure gas chromatographic relative retention time models on seven stationary phases for 209 polybrominated diphenyl ether congeners. AB - Quantitative structure retention relationships (QSRRs) were developed to predict the gas chromatographic (GC) relative retention times (RRTs) for 209 polybrominated diphenyl ether (PBDE) congeners using the heuristic method included in the computer software Comprehensive Descriptors for Structural and Statistical Analysis (CODESSA). A total of 445 constitutional, topological, geometrical, electrostatic, and semi-empirical quantum chemical descriptors were derived for all PBDEs. Using experimental RRT data for 126 PBDE congeners from the literature, predictive regression models were built for seven individual GC capillary columns differing in stationary phases. Each model includes four descriptors which included Wiener index, Randic index, polarity parameter, etc., selected by CODESSA. High predictability was obtained. High multiple correlation coefficients R(2) indicated that >98.5% (except for stationary phase CP-Sil 19) of the total variation in the predicted RRTs is explained by the fitted models. The models were subsequently used to predict the RRTs of the remaining 83 PBDE congeners on seven different stationary phases. The statistical results show that, compared with others, DB-XLB column not only produces the least number of peak overlaps but also results in shorter retention times. PMID- 16352310 TI - Brain SPECT analysis by 3D-SSP and phenotype of Parkinson's disease. AB - OBJECTIVES: We hypothesize that the regional pattern of blood flow reduction in the brain is different between tremor-dominant Parkinson's disease (PD) and postural instability gait difficulty (PIGD)-dominant PD. We therefore investigated the association of phenotypes in untreated PD with brain perfusion on SPECT using three-dimensional stereotactic surface projection (3D-SSP) technique. PATIENTS AND METHODS: Thirty-three patients who had PD without dementia (12 men and 21 women with a mean age of 67.1+/-6.4 years) were included in this study. Their symptoms were rated using the Unified Parkinson's Disease Rating Scale (UPDRS). Patients were grouped in two phenotypes: tremor and PIGD dominant groups based on UPDRS components. Around the same time, all patients were examined by N-isopropyl-p[123I] iodoamphetamine single photon emission computed tomography (123I-IMP SPECT), and obtained images were analyzed with 3D SSP using an image-analysis software, NEUROSTAT. Data on brain surface perfusion extracted by 3D-SSP analysis were compared between the PD patients and the normal control group. The same comparisons were made for subgroups of PD patients. RESULTS: Cerebral perfusion was decreased at the anterior cingulate cortex and primary visual cortex of the PD patients, and especially by the pixel-by-pixel comparison, perfusion was significantly decreased at the right anterior cingulate cortex compared with the normal controls. In the PIGD-dominant group, more severe hypoperfusion was seen at the same regions. In the tremor-dominant group, significant hypoperfusion was not seen compared with the normal controls. CONCLUSIONS: The regional pattern of blood flow reduction in the brain was found to be different between tremor-dominant PD and PIGD-dominant PD. These regional differences were considered to suggest different and disease-specific combinations of underlying pathophysiological and neurochemical processes. PMID- 16352311 TI - Ceruloplasmin and superoxide dismutase (SOD1) in Parkinson's disease: a follow-up study. AB - In this follow-up study concentration, oxidative activity and specific oxidative activity of ceruloplasmin (CP) in serum and the activity of superoxide dismutase (SOD1) in erythrocytes were reexamined in 28 of originally 40 patients with Parkinson's disease (PD), and their age- and gender-matched controls. The mean CP and SOD1 parameters were significantly lower in the patients than in the controls. SOD1 activity and age of the patients were inversely correlated. The patients were divided into two subgroups based on their H&Y score i.e. groups II and III (12 patients) versus groups IV and V (16 patients). No significant difference was found in the CP or SOD1 parameters between the subgroups. Patients were also divided into two subgroups based on treatment with levodopa and decarboxylase blocker alone (12 patients) or given additionally a dopamine agonist (15 patients). No significant difference in the parameters was found between these subgroups in relation to intake of dopamine agonists. Results of this study are in agreement with results of the former study 5 years earlier. There is considerable overlap in individual values between patients and controls of the parameters studied. Thus CP and SOD1 have no obvious value for diagnosis or clinical evaluation of PD. PMID- 16352312 TI - Dysautonomic achalasia in two siblings with Sandhoff disease. AB - Two siblings in their sixth decade with chronic Type II GM2 gangliosidosis developed progressive dysphagia in addition to chronic motor neuron disease and autonomic nervous system (ANS) involvement. Esophageal achalasia was diagnosed in both patients. It is suggested that this esophageal motor disorder may be a manifestation of the neurovegetative system disorder due to alteration of ganglioside metabolism. PMID- 16352313 TI - Computer modelling of antifolate inhibition of folate metabolism using hybrid functional petri nets. AB - Antifolates are used in the treatment of various human malignancies and exert their cytotoxic activity by inhibiting folate-dependent enzymes resulting in disruption of DNA synthesis and cell death. Here we devised a computerized hybrid functional petri nets (HFPN) modelling of folate metabolism under physiological and antifolate inhibitory conditions. This HFPN modelling proved valid as a good agreement was found between the simulated steady-state concentrations of various reduced folates and those published for cell extracts; consistently, the simulation derived total folate pool size (11.3 microM) was identical to that published for cell extracts. In silico experiments were conducted to characterize the inhibitory profile of four distinct antifolates including methotrexate (MTX), tomudex, and LY309887, which inhibit dihydrofolate reductase (DHFR), thymidylate synthase (TS) and glycineamide ribonucleotide transformylase (GARTFase), respectively, as well as pemetrexed which has the capacity to inhibit all three enzymes. In order to assess the inhibitory activity of antifolates on purines and pyrimidines, the biosynthesis rates of IMP (20.53 microM/min) and dTMP (23.8 microM/min) were first simulated. Whereas the biochemical inhibitory profile of MTX was characterized by increased dihydrofolate and decreased tetrahydrofolate (THF) concentrations, the remaining antifolates did not decrease THF levels. Furthermore, MTX was 766- and 10-fold more potent in decreasing the production rates of IMP and dTMP, respectively, than pemetrexed. LY309887 indirectly decreased the rate of dTMP production by reducing the levels of 5-CH2-THF, a folate cofactor for TS. Surprisingly, pemetrexed failed to inhibit DHFR even at high concentrations. This HFPN-based simulation offers an inexpensive, user friendly, rapid and reliable means of pre-clinical evaluation of the inhibitory profiles of antifolates. PMID- 16352314 TI - A biomechanical model for size, speed and anatomical variations of the energetic costs of running mammals. AB - Here we propose a model of energetic costs and the muscle-tendon unit function on running mammals. The main goal is to set a simple theoretical framework which gives an understanding of the biomechanical principles behind the size, speed and anatomical variations of the energetic costs of running mammals. The model is a point-like mass withstood by a two-segment leg with an extensor muscle serially attached to a tendon. We considered withstanding body weight during the stance phase as the main role of the muscle-tendon unit during fast locomotion. The ground reaction force dependence on speed and the time of stance phase as well as other biomechanical characteristics were taken from previous empirical studies of running. At the same time, the morphological variations with body mass were taken from empirically well-established allometric equations for mammals. The metabolic cost was estimated from an empirical equation relating metabolic power with muscular force and speed in shortening and stretching. Our model predicts the pattern of mass specific metabolic rate variations with both speed and body mass. It also gives an explanation of the experimentally reported linear inverse relationship between the rate of energy used for running and the time of application of force by the foot to the ground during each stride. It also suggests an explanation of the unusual energy saving adaptations of large macropodids. It provides some predictions on the relationship, between energy costs and muscle-tendon unit characteristics, testable on further experiments. PMID- 16352315 TI - Modeling the onset of drug dependence: a consideration of the requirement for protein synthesis. AB - It has been proposed, with some supporting evidence, that development of opiate tolerance and dependence requires protein synthesis. However, a quantitative, biologically based model within which to analyse and support the data has been lacking. Utilizing such a framework or model, we recently compared the time course of onset of opiate dependence in laboratory animals, with the mathematical time course of general changes in protein levels. Not only did the time course of onset of dependence parallel the time course of increasing levels of a protein, but also the half-life of the putative protein required by the model was very similar to those of many brain proteins. In this study, we have more extensively tested the model by producing and examining a much more detailed and surprisingly complex time course of the onset of dependence. Applying the protein synthesis time course model to the data suggested the presence of two distinct components of dependence, an early transient component and a later long-lasting component. These components appear to correspond to acute and chronic dependence, respectively. The protein synthesis hypothesis more readily applies to the chronic dependence portion. Because consideration of the model can generate components that correspond to accepted and well-known components of dependence, both the utility of the model as well as the hypothesis that opiate dependence at least partially requires protein synthesis are supported. It is also possible that individual components of the withdrawal syndrome have individual and unique rate limiting mechanisms. In any case, time course analysis may be helpful in revealing underlying mechanisms of change. PMID- 16352316 TI - Spatial effects in social dilemmas. AB - Social dilemmas and the evolutionary conundrum of cooperation are traditionally studied through various kinds of game theoretical models such as the prisoner's dilemma, public goods games, snowdrift games or by-product mutualism. All of them exemplify situations which are characterized by different degrees of conflicting interests between the individuals and the community. In groups of interacting individuals, cooperators produce a common good benefitting the entire group at some cost to themselves, whereas defectors attempt to exploit the resource by avoiding the costly contributions. Based on synergistic or discounted accumulation of cooperative benefits a unifying theoretical framework was recently introduced that encompasses all games that have traditionally been studied separately (Hauert, Michor, Nowak, Doebeli, 2005. Synergy and discounting of cooperation in social dilemmas. J. Theor. Biol., in press.). Within this framework we investigate the effects of spatial structure with limited local interactions on the evolutionary fate of cooperators and defectors. The quantitative effects of space turn out to be quite sensitive to the underlying microscopic update mechanisms but, more general, we demonstrate that in prisoner's dilemma type interactions spatial structure benefits cooperation although the parameter range is quite limited-whereas in snowdrift type interactions spatial structure may be beneficial too, but often turns out to be detrimental to cooperation. PMID- 16352317 TI - Vanadium inhibits the development of 2-acetylaminofluorene-induced premalignant phenotype in a two-stage chemical rat hepatocarcinogenesis model. AB - In recent years, research on the biological influence of micronutrients in cancer has grown enormously. Among these, vanadium, a dietary micronutrient present in mammalian tissues has received considerable attention as a limiting agent. In the present study, attempts have been made to investigate the in vivo antitumour potentials of this micronutrient at the 0.5 ppm dosage in drinking water in a defined model of a two-stage experimental rat hepatocarcinogenesis. The chemopreventive effect of vanadium was assessed by studying certain biomarkers, such as development of gamma-glutamyltranspeptidase (GGT)-positive foci, levels of some essential trace elements, in situ expression of proliferating cell nuclear antigen (PCNA) and chromosomal aberrations. Hepatocarcinogenesis was induced in male Sprague-Dawley rats by chronic feeding of 2-acetylaminofluorene (0.05% in basal diet) on and from week 4. Vanadium administration throughout the experiment reduced the relative liver weight, nodular incidence (66.70%), total number and multiplicity (79.93%) and restored hepatic levels of selenium (Se) and iron (Fe) (P < 0.001) when compared to the carcinogen control. Moreover, long term vanadium treatment significantly abated the expressions of GGT (P < 0.001) and PCNA with concomitant reduction in PCNA immunolabeling index (P < 0.001; 36.62%). Finally, the anticlastogenic potential of vanadium was reflected through its ability to inhibit early chromosomal aberrations (P < 0.001; 45.17%) in 2-AAF challenged rat hepatocytes. Our results suggest that supplementary vanadium at a dose of 0.5 ppm, when administered continuously throughout the study, than administered either in the initiation or promotion phase alone, is very much effective in suppressing neoplastic transformation in vivo. We conclude the significant role of vanadium in limiting cell proliferation and chromosomal aberrations during the preneoplastic stages of hepatocarcinogenesis in rats. PMID- 16352318 TI - Anti-oxidative effect of triterpene acids of Eriobotrya japonica (Thunb.) Lindl. leaf in chronic bronchitis rats. AB - The study was to evaluate the effect of triterpene acids of Eriobotrya japonica (Thunb.) Lindl. leaf (TAL) on expression of antioxidative mediators by alveolar macrophages (AM) in rats with chronic bronchitis (CB), CB was induced by endotracheal instillation of lipopolysaccharedes (LPS) followed by Bacillus Calmette-Guerin (BCG) injection through caudal vein 1 week later. Treatment groups received TAL at there different doses (50, 150, or 450 mg/kg daily, intragastrically (i.g.)) or dexamethasone (1.2 mg/kg daily i.g.) for 2 weeks, 7 days after LPS injection. AM were then isolated and incubated. Superoxide dismutase (SOD) and methylene dianiline (MDA) levels in AM were measured by commercial kits; meanwhile, heme oxygenase-1 (HO-1) expression and its mRNA expression in AM were detected by immunocytochemistry and RT-PCR, respectively. HO-1 activity of the lung was also detected by a specific biochemistry reaction. The levels of MDA and HO-1 expressed by cultured AM and the HO-1 activity in the lung of the TAL groups were significantly lower than those from the CB group without treatment (p < 0.01 and p < 0.05, respectively), while the SOD levels were increased in a dose-dependent manner by TAL treatment. These results suggest that TAL inhibits HO-1 expression and MDA production and up-regulates SOD expression in AM from CB rats, which might be one of molecular mechanisms of its anti-inflammatory effects in CB rats. PMID- 16352319 TI - The role of semantic distance in category-specific impairments for living things: evidence from a case of semantic dementia. AB - In this paper, we describe a patient (LI) suffering from semantic dementia who showed a category-specific naming impairment for living things over and above the effects of several nonsemantic confounding variables. We investigated the characteristics of LI's impairment to address the following three issues raised in three different accounts of category-specific impairments: (i) the role of an imbalance in the loss of sensory compared to nonsensory features (assumed by the Sensory Functional Theory [Warrington, E. K., & Shallice, T. (1984). Category specific semantic impairments. Brain, 107, 829-859]); (ii) the role of cross domain differences in Feature Correlation (assumed by the Conceptual Structure Account [Moss, H., Tyler, L. K., & Devlin, J. T. (2002). The emergence of category-specific deficits in a distributed semantic system. In: E. M. E. Forde & G. W. Humphreys (Eds.), Category Specificity in Brain and Mind (pp. 115-147). New York: Psychology Press]); (iii) the role of semantic distance (proposed by Cree and McRae [Cree, G. S., & McRae, K. (2003). Analyzing the factors underlying the structure and computation of the meaning of chipmunk, cherry, chisel, cheese, and cello (and many other such concrete nouns). Journal of Experimental Psychology: General, 132, 163-201]). We found that semantic distance was the only factor causally linked to LI's poorer performance on living things. In fact, her naming performance was less accurate on items that had many semantic neighbours, which is typical of living things. On the contrary, a feature listing task revealed that the features available to LI were not predicted by their level of correlation, as expected by the Conceptual Structure Account. Finally, at variance with the Sensory Functional Theory, although LI quoted sensory features less accurately than nonsensory ones, this did not give rise to a disproportionate loss of semantic features in the living domain. PMID- 16352320 TI - Hippocampal head atrophy after traumatic brain injury. AB - Traumatic brain injury (TBI) causes hippocampal damage. The hippocampus can be macroscopically divided into the head, body and tail, which differ in terms of their sensitivity to excitability and also in terms of their cortical connections. We investigated whether damage also varies according to the hippocampal area involved, and studied the relationship of hippocampal reductions with memory performance. Twenty TBI patients and matched controls were examined. MRI measurements were performed separately for the hippocampal head, body and tail. Memory outcome was measured by Rey's auditory verbal learning test, Rey's complex figure test and a modified version of Warrington's facial recognition memory test. Group comparison showed that patients had bilateral hippocampal atrophy, mainly involving the hippocampal head. Moreover, TBI subjects showed verbal memory deficits which presented slight correlations with left hippocampal head atrophy. PMID- 16352321 TI - Failure to read motor intentions from gaze in children with autism. AB - A core feature of autism is the abnormal use of gaze to attribute mental states to others, and thus to predict others' behaviour. An untested idea is whether this dysfunction is confined to mental states having a propositional content, such as beliefs and desire or extends to motor intentional states which allow one to make inferences about the actions of others. This study used kinematics to examine the ability to use gaze to inform one about the motor states of another in normal and autistic children. In each trial two participants, a model and an observer, were seated facing each other at a table. In three experimental blocks the model was requested to grasp a stimulus, to gaze towards the same stimulus, and to gaze away from the stimulus without performing any action. The task for the observer was to grasp the stimulus after having watched the model perform her task. We observed that normal children showed facilitation effects in terms of movement speed following the observation of the model grasping or simply gazing at the object. In contrast, autistic children did not show any evidence of facilitation in these conditions. Neither normal nor autistic children showed evidence of facilitation when the model's gaze was not directed towards the stimulus. These findings demonstrate that, in contrast to normal children, children with autism fail to use information from the model's action or gaze to plan their subsequent action, and that in autism the inability to use of another person's gaze produces a lack of understanding of the motor intention of others. PMID- 16352322 TI - Effects of sibutramine on the appetitive and consummatory aspects of feeding in non-human primates. AB - This study examined how sibutramine (0.06-4.0 mg/kg, i.m.), a clinically effective weight-loss medication which increases extracellular serotonin and norepinephrine levels, affected the appetitive and consummatory aspects of feeding of non-human primates. The effects were compared to the effects of the positive control dexfenfluramine (2.0-6.0 mg/kg, p.o.), which primarily increases extracellular serotonin levels. Baboons had access to food 24 h each day, but they had to complete a two-phase operant procedure in order to eat. Responding on one lever during a 30-min appetitive phase was required before animals could start a consumption phase, where responding on another lever led to food delivery, i.e., a meal. Responding during the appetitive phase resulted in presentations of food-related stimuli only. Sibutramine increased the latency to the first meal of the session in females, and decreased consummatory behavior without affecting other appetitive behavior in males and females. In contrast, dexfenfluramine, increased the latency to the first meal of the session, and decreased both appetitive and consummatory behavior in males and females. The behavioral mechanism by which sibutramine decreases food intake is distinct from other anorectic drugs, including dexfenfluramine, that have been tested in this paradigm. PMID- 16352323 TI - Abomasal bacteria produce an inhibitor of gastrin secretion in vitro. AB - Previously, proliferating microflora transferred with abomasal nematodes, were suspected to be the source of the gastrin inhibitor in some parasite excretory/secretory products. Aerobic cultures in HBSS of abomasal fluid from uninfected sheep became inhibitory during the static growth phase, unless antibiotics were present. Basal gastrin secretion was reduced by up to 90%. Rumen fluid and incubates and medium in which Streptococcus bovis and ovine rumen Actinomycete spp. had been grown also contained the inhibitor. Unlike abomasal cultures, rumen fluid and incubates also reduced the measurement of gastrin standards. Rumen incubates were less potent after exposure to pH 2-3, suggesting that inactivation normally occurs in the unparasitised abomasum. Contaminating bacteria which generate the gastrin inhibitor in parasite ES products are probably rumen organisms which survive in the abomasum and proliferate during subsequent incubation. Significantly, rumen bacteria have been shown to be capable of affecting the secretory activity of the gastric mucosa. PMID- 16352324 TI - Complex genetic structure of the rabies virus in Bangkok and its surrounding provinces, Thailand: implications for canine rabies control. AB - Dog vaccination and population management have been suggested as priorities in attempts at disease control in canine rabies-endemic countries. Budget limitations and the complexity of social, cultural and religious variables have complicated progress in the developing world. In Bangkok, Thailand, an intensive canine vaccination and sterilization programme has been in place since November 2002. Our objective was to determine if the rabies virus could be mapped according to its genetic variations and geographical location on the small localized scale of Bangkok and its surrounding provinces. Phylogenetic characterization of 69 samples from Bangkok and five neighbouring and two remote provinces, by limited sequence analysis of the rabies virus nucleoprotein gene, distinguished six different clades. Rabies viruses of four clades were intermixed in Bangkok and in the surrounding highly populated regions whereas the other two clades were confined to rural and less populated provinces. Such a complex pattern of gene flow, particularly in Bangkok, may affect the outcome of canine control programmes. PMID- 16352325 TI - A chemical signal possibly related to physiology in fossil cells detected by energy dispersive X-ray microanalysis. AB - Energy dispersive X-ray microanalysis (EDXMA) is a widely used tool employed to detect elemental composition and its spatial distribution in a sample without causing damage. Charcoalified cytoplasm is a new type of fossil material that came to people's attention only recently. In this paper, EDXMA is used for the first time to detect the spatial elemental distribution in charcoalified cytoplasm of two fossil plants that are more than 100 million years old. The results demonstrate certain elemental distribution patterns within charcoalified cytoplasm and the surrounding cell walls. Based on the results from cytological studies of extant material, the heterogeneous spatial elemental distribution within the charcoalified cytoplasm has the potential to be related to the maturation of cells, the presence of certain organelles, and the physiology of these organelles. This is the first chemical signal detected in cytoplasm residue that can possibly be related to plant physiology. This paves the way for further research on fossil cytoplasm, which will better our understanding on the physiology of fossil plants. PMID- 16352326 TI - Chlorpromazine reduces the intercellular communication via gap junctions in mammalian cells. AB - In the work presented herein, we evaluated the effect of chlorpromazine (CPZ) on gap junctions expressed by two mammalian cell types; Gn-11 cells (cell line derived from mouse LHRH neurons) and rat cortical astrocytes maintained in culture. We also attempted to elucidate possible mechanisms of action of CPZ effects on gap junctions. CPZ, in concentrations comparable with doses used to treat human diseases, was found to reduce the intercellular communication via gap junctions as evaluated with measurements of dye coupling (Lucifer yellow). In both cell types, maximal inhibition of functional gap junctions was reached within about 1 h of treatment with CPZ, an recovery was almost complete at about 5 h after CPZ wash out. In both cell types, CPZ treatment increased the phosphorylation state of connexin43 (Cx43), a gap junction protein subunit. Moreover, CPZ reduced the reactivity of Cx43 (immunofluorescence) at cell interfaces and concomitantly increased its reactivity in intracellular vesicles, suggesting an increased retrieval from and/or reduced insertion into the plasma membrane. CPZ also caused cellular retraction reducing cell-cell contacts in a reversible manner. The reduction in contact area might destabilize existing gap junctions and abrogate formation of new ones. Moreover, the CPZ-induced reduction in gap junctional communication may depend on the connexins (Cxs) forming the junctions. If Cx43 were the only connexin expressed, MAPK-dependent phosphorylation of this connexin would induce closure of gap junction channels. PMID- 16352327 TI - Arsenic removal from high-arsenic water by enhanced coagulation with ferric ions and coarse calcite. AB - Arsenic removal from high-arsenic water in a mine drainage system has been studied through an enhanced coagulation process with ferric ions and coarse calcite (38-74 microm) in this work. The experimental results have shown that arsenic-borne coagulates produced by coagulation with ferric ions alone were very fine, so micro-filtration (membrane as filter medium) was needed to remove the coagulates from water. In the presence of coarse calcite, small arsenic-borne coagulates coated on coarse calcite surfaces, leading the settling rate of the coagulates to considerably increase. The enhanced coagulation followed by conventional filtration (filter paper as filter medium) achieved a very high arsenic removal (over 99%) from high-arsenic water (5mg/l arsenic concentration), producing a cleaned water with the residual arsenic concentration of 13 microg/l. It has been found that the mechanism by which coarse calcite enhanced the coagulation of high-arsenic water might be due to attractive electrical double layer interaction between small arsenic-borne coagulates and calcite particles, which leads to non-existence of a potential energy barrier between the heterogeneous particles. PMID- 16352328 TI - Identity and distribution of residues of energetic compounds at army live-fire training ranges. AB - Environmental investigations have been conducted at 23 military firing ranges in the United States and Canada. The specific training facilities most frequently evaluated were hand grenade, antitank rocket, and artillery ranges. Energetic compounds (explosives and propellants) were determined and linked to the type of munition used and the major mechanisms of deposition. PMID- 16352329 TI - Oxidative ring cleavage of low chlorinated biphenyl derivatives by fungi leads to the formation of chlorinated lactone derivatives. AB - The yeast Trichosporon mucoides and the filamentous fungus Paecilomyces lilacinus as biphenyl oxidizing organisms are able to oxidize chlorinated biphenyl derivatives. Initial oxidation of derivatives chlorinated at C4 position started at the non-halogenated ring and went on up to ring cleavage. The products formed were mono- and dihydroxylated 4-chlorobiphenyls, muconic acid derivatives 2 hydroxy-4-(4-chlorophenyl)-muconic acid and 2-hydroxy-5-(4-chlorophenyl)-muconic acid as well as the corresponding lactones 4-(4-chlorophenyl)-2-pyrone-6 carboxylic acid and 3-(4-chlorophenyl)-2-pyrone-6-carboxylic acid. Altogether T. mucoides formed 12 products and P. lilacinus accumulated five products. Whereas the rate of the first oxidation step at 4-chlorobiphenyl seems to be diminished by the decreased bioavailability of the compound, no considerable differences were observed between the degradation of 4-chloro-4'-hydroxybiphenyl and 4 hydroxybiphenyl. Twofold chlorinated biphenyl derivatives did not serve as substrates for oxidation by either organism with the exception of 2,2' dichlorobiphenyl, transformed by the yeast Trichosporon mucoides to two monohydroxylated derivatives. The results show, that soil fungi may contribute to the aerobic degradation of low chlorinated biphenyls accumulating from anaerobic dehalogenation of PCB by bacteria. PMID- 16352331 TI - Accumulation of memory T cells from childhood to old age: central and effector memory cells in CD4(+) versus effector memory and terminally differentiated memory cells in CD8(+) compartment. AB - Memory T cells can be classified as central memory (T(CM), CD45RA(neg)CCR7(+)), effector memory (T(EM), CD45RA(neg)CCR7(neg)), and terminally differentiated cells (T(TD), CD45RA(+)CCR7(neg)) with different homing and effector capacities. In 101 healthy subjects aged from 5 to 96 years, distinct dynamics were evidenced between circulating CD4(+) and CD8(+) T cell populations. Naive CD4(+) and CD8(+) T cells decreased linearly with age, CD8(+) twice more rapidly. Memory cells outnumbered naive cells on average at 37.4 in the CD4(+) and 29.5 years of age in the CD8(+) pool. CD4(+) T(CM) and T(EM) cells were positively correlated and increased linearly at a similar rate with age, while CD4(+) T(TD) remained rare. CD8(+) T(EM) and T(TD) accumulated linearly with age, while T(CM) increased only slightly, and each memory subset was negatively correlated to the two others. Almost all CD8(+) T(TD) and some CD8(+) T(EM) had lost CD28 expression. Despite different dynamics, each individual CD4(+) naive and memory subset was correlated to the synonymous CD8(+) subset. Half of the subjects aged 65 years or older were characterized by extremely reduced CD8(+) naive and increased CD8(+) T(TD) cell counts, which could indicate an acceleration of the decay of the immune system from this age onward. PMID- 16352330 TI - Factors influencing lead sorption-desorption at variable added metal concentrations in Rhodoxeralfs. AB - The response of ten soils of the lithic Rhodoxeralf type to the supply of lead at concentrations of 500, 1000, 2000, 3000, 4000, 5000 and 6000 mg kg-1 was examined in batch sorption-desorption tests. Lead availability in the soils was found to depend on its partitioning between the soil solution and the solid phase as reflected in adsorption isotherms. The isotherms, of the H type, were consistent with a high affinity of the sorbent for the metal, with which it forms stable inner-sphere complexes on the soil surface. Sorption-desorption tests revealed that some properties of the soils such as their pH (mean=8) and high contents in clays (particularly in kaolinite) and crystalline iron oxides significantly influence Pb sorption, the effect being especially marked at high added metal concentrations. Added lead is largely retained by crystalline iron oxides and the soil clay fraction; the pH of the soil favours the release of variably-charged sites from both. The extent of Pb desorption was small, particularly at the lowest added levels (500 and 1000 mg kg-1). Desorption increased with increasing added Pb concentration and exceeded 50% at 5000 and 6000 mg kg-1; this suggests that Pb is present not only as inner-sphere complexes, but also as outer-sphere complexes and, partly, as precipitates. The desorption isotherms consist of three segments that exhibit significant differences depending on the added Pb concentration, namely: 500-1000, 2000-4000 and 5000-6000 mg kg-1. PMID- 16352332 TI - Protein Z is not synthesised by human umbilical vein endothelial cells. PMID- 16352333 TI - Comparison of uterine malignancies that develop during and following tamoxifen therapy. AB - OBJECTIVES: There is a greater than 7-fold increased risk of uterine cancer in women with breast cancer exposed to tamoxifen. The objective of this study was to determine the percentage of women who developed uterine cancer more than 12 months after discontinuing tamoxifen (past users) and to compare their clinical and pathologic features with those of women who developed uterine cancer while on tamoxifen therapy or within 12 months of stopping therapy (recent users). METHODS: All women with a diagnosis of uterine cancer at Memorial Sloan-Kettering Cancer Center between 1980 and June 2004 with a past history of breast cancer treated with tamoxifen were identified. Clinical and pathologic data were obtained through retrospective chart review. RESULTS: There were 106 women identified with a history of breast cancer treated with tamoxifen preceding their diagnosis of uterine cancer. Thirty-nine (37%) developed uterine cancer more than 12 months after discontinuing tamoxifen. The median time until developing uterine cancer in past users was 33 months (range, 13-22). There were no significant differences in age at breast cancer diagnosis, body mass index, parity, stage of breast cancer, modality of breast cancer treatment, or duration of tamoxifen therapy between past and recent users of tamoxifen. Women who were past users of tamoxifen had significantly more FIGO (International Federation of Gynecology and Obstetrics) grade 3 and non-endometrioid histologic subtypes (P = 0.009; P = 0.007). CONCLUSIONS: More than one third of women treated with tamoxifen develop uterine cancer more than 12 months after discontinuing therapy. These women are at greater risk of developing moderately to poorly differentiated tumors, which is a known poor prognostic factor. Therefore, women with a past history of tamoxifen therapy should have continued surveillance after completion of tamoxifen to ensure early diagnosis of uterine cancer. PMID- 16352334 TI - Influence of environmental conditions on dental composite flexural properties. AB - OBJECTIVE: To determine if increased relative humidity and temperature simulating intraoral environmental conditions adversely affect flexural properties of dental composites. METHODS: Specimen fabrication followed ANSI/ADA specification 27 for resin-based filling materials, except that ambient laboratory conditions (47% relative humidity at 22 degrees C) or simulated intraoral conditions (90% relative humidity at 35 degrees C) were used when fabricating and polymerizing specimens. Ten specimens were made of each of three commercially available composites at each condition. As per the specification, after aging specimens in 10 ml of deionized water at 37 degrees C for 24 h, flexural properties were measured using a 3-point bend test. RESULTS: A two-factor ANOVA and Fisher's least significant difference (LSD) post hoc (alpha=0.05) indicated there were significant differences in flexural modulus and strength as a function of material, with Z250=TPH>Prodigy. However, neither flexural modulus nor flexural strength of any material was adversely influenced by fabrication conditions. SIGNIFICANCE: Although the flexural properties did not decrease with respect to fabrication conditions, the flexural modulus of some of the materials (TPH Spectrum and Z250) increased when specimens were fabricated at simulated intraoral temperature and relative humidity. Thus, simulation of these factors may be important in laboratory testing, since the resultant properties may better reflect flexural properties associated with dental composite restorations placed clinically. PMID- 16352335 TI - Electrospinning polyaniline-contained gelatin nanofibers for tissue engineering applications. AB - Polyaniline (PANi), a conductive polymer, was blended with a natural protein, gelatin, and co-electrospun into nanofibers to investigate the potential application of such a blend as conductive scaffold for tissue engineering purposes. Electrospun PANi-contained gelatin fibers were characterized using scanning electron microscopy (SEM), electrical conductivity measurement, mechanical tensile testing, and differential scanning calorimetry (DSC). SEM analysis of the blend fibers containing less than 3% PANi in total weight, revealed uniform fibers with no evidence for phase segregation, as also confirmed by DSC. Our data indicate that with increasing the amount of PANi (from 0 to approximately 5%w/w), the average fiber size was reduced from 803+/-121 nm to 61+/-13 nm (p<0.01) and the tensile modulus increased from 499+/-207 MPa to 1384+/-105 MPa (p<0.05). The results of the DSC study further strengthen our notion that the doping of gelatin with a few % PANi leads to an alteration of the physicochemical properties of gelatin. To test the usefulness of PANi-gelatin blends as a fibrous matrix for supporting cell growth, H9c2 rat cardiac myoblast cells were cultured on fiber-coated glass cover slips. Cell cultures were evaluated in terms of cell proliferation and morphology. Our results indicate that all PANi-gelatin blend fibers supported H9c2 cell attachment and proliferation to a similar degree as the control tissue culture-treated plastic (TCP) and smooth glass substrates. Depending on the concentrations of PANi, the cells initially displayed different morphologies on the fibrous substrates, but after 1 week all cultures reached confluence of similar densities and morphology. Taken together these results suggest that PANi-gelatin blend nanofibers might provide a novel conductive material well suited as biocompatible scaffolds for tissue engineering. PMID- 16352336 TI - N-halamine/quat siloxane copolymers for use in biocidal coatings. AB - A series of copolymers incorporating N-halamine siloxane and quaternary ammonium salt siloxane units has been prepared. The primary function of the quat units was to render the siloxane copolymers soluble in water. The copolymers have been coated onto cotton swatches and evaluated for biocidal efficacy against Staphylococcus aureus and Escherichia coli O157:H7. It was determined that both N halamine and quat functional groups were effective against S. aureus, but only the N-halamine units were effective against Escherichia coli O157:H7. The copolymers should be useful for applications for which aqueous media is preferred over organic solvents to be used during coating procedures. PMID- 16352337 TI - Effects of linear cationic alpha-helical antimicrobial peptides on immune relevant genes in trout macrophages. AB - There are increasing evidence of the potential role of antimicrobial peptides in the regulation of immune responses in mammalian species. However, the effects of these peptides in fish have yet to be investigated. In this study, we examined the transcriptional expression profile of representative immune-relevant genes in a trout macrophage cell line, RTS11, in response to three linear cationic alpha helical antimicrobial peptides (insect cecropin B, fish pleurocidin and a cecropin analogue CF17). The expression levels of two pro-inflammatory genes, interleukin-1 beta (IL-1 beta) and cyclo-oxygenase-2 (COX-2), increased in the peptide-treated RTS11 cells. The peptides did not appear to affect the expression levels of representative genes associated with antigen presentation, interferon response or JAK/STAT signal transduction. Furthermore, the induction of IL-1 beta and COX-2 in RTS11 by lipopolysaccharide was not adversely affected by these three antimicrobial peptides. Overall, the data indicate a pro-inflammatory effect of the three cationic antimicrobial peptides in the inflammatory response of salmonid species, suggesting a potential application of these peptides as immune adjuvant for fish vaccination. PMID- 16352338 TI - Haematological malignancies--a predictor of a poor outcome in patients with bacteraemia. AB - OBJECTIVES: To compare microbial agents causing first episodes of bacteraemia in patients with and without haematological malignancies, and to examine mortality within 90 days of follow-up. METHODS: We conducted a population-based study in North Jutland County, Denmark including patients (> or = 15 years) registered in the County Bacteraemia Registry with their first episode of bacteraemia during the period 1992-2002. Diseases prior to the bacteraemia were identified in the Hospital Discharge Registry. Mortality was determined through the Civil Registration System. We estimated adjusted mortality rate ratios (MRRs) using Cox's regression analysis. RESULTS: We included 7456 patients with a first episode of bacteraemia; of these, 444 (6%) had a haematological malignancy. Haematological patients had almost the same proportions of gram-negative (49% vs. 50%) and gram-positive (37% vs. 39%) bacteraemia as other patients. Cumulative 30 day mortality in patients with haematological malignancies was 32% vs. 20% in patients without any malignancy, yielding an adjusted MRR of 1.6 (95% CI 1.3 2.0). For patients with non-haematological malignancies the adjusted MRR was 1.5 (95% CI 1.4-1.7). CONCLUSION: Despite a similar distribution of the microbial agents causing bacteraemia, patients with haematological malignancies and also patients with non-haematological malignancies had increased 30-day mortality of bacteraemia when compared with patients without any malignancy. PMID- 16352339 TI - Severe pneumonia during primary infection with an atypical strain of Toxoplasma gondii in an immunocompetent young man. AB - We report a case of severe acute primary pulmonary toxoplasmosis in an immunocompetent young man living in Nice (Southern France). The Toxoplasma DNA extracted from the broncho-alveolar lavage fluid allowed a genetic characterization of the responsible strain which displayed an atypical genotype of Toxoplasma gondii. This unusual genetic composition of the parasite may have influenced, among other factors, the severity of the disease. PMID- 16352340 TI - Chronic Chlamydia pneumoniae infection in patients with symptomatic atherothrombosis. AB - OBJECTIVES: The aim of the present study was to search for an association between chronic Chlamydia pneumoniae infection, indicated by elevated antibody titers against the pathogen, atherothrombosis and the occurrence of arterial ischemic events. METHODS: We studied 52 patients presenting at baseline with at least one symptomatic episode of atherothrombosis. A screening for fasting blood glucose and a lipid profile was performed on all patients who had no known history of diabetes or hypercholesterolemia. RESULTS: The prevalence of IgG and IgA anti-C. pneumoniae antibodies at baseline was 90% (95% CI: 79-97) and 81% (67-90), respectively. Forty-two of the 52 patients (81%) experienced a new arterial ischemic event after a mean follow-up of 9 years [heart: 19 (37%); brain: 12 (23%); lower limbs: 8 (15%); and other: 13 (25%)]. Occurrence of a new arterial ischemic event was related to age (p=0.003), sex (p=0.009), and tobacco smoking (p=0.06). Prevalences of IgA and IgG anti-C. pneumoniae were significantly higher in patients with atherothrombosis at baseline than that in controls. CONCLUSION: Our study confirmed the links between C. pneumoniae and atherothrombosis. However, neither IgA nor IgG antibodies for C. pneumoniae was a significant predictive factor for new ischemic arterial events in patients with atherothrombosis. PMID- 16352341 TI - DRESS like severe drug rash with eosinophilia, atypic lymphocytosis and fever secondary to ceftriaxone. AB - We report a case in a 50-year-old male who had been treated with ceftriaxone for 3 weeks to treat meningitis. He was admitted 4 days after cessation of the ceftriaxone treatment with fever, headache, nausea, vomiting, myalgia, arthralgia, pruriginous skin rashes, and with edema on face. Blood tests showed marked eosinophilia and atypic lymphocytosis. He was successfully treated with prednisone p.o. We report this case as we could not recognize a case like this which was induced by ceftriaxone. PMID- 16352342 TI - The incidence of Bordetella pertussis infections estimated in the population from a combination of serological surveys. AB - OBJECTIVES: Bordetella pertussis circulates even in highly vaccinated populations. There is a considerable amount of infection in adults. For designing more effective vaccination schedules it is important to quantify the age dependent relation between the number of notified cases and the number of infections. METHODS: We used a statistical relationship between the time since infection and the IgG antibody titers against pertussis toxin, derived from a longitudinal data set, to estimate time since infection for all individuals in a cross-sectional population-based study (1995-1996) based on their titers. Age specific incidence of infection with B. pertussis was calculated and compared with the age-distribution of notified cases of pertussis in 1994-1996. RESULTS: Estimated incidence of infection was 6.6% per year for 3-79-year olds, annual incidence of notified cases 0.01%. Estimated age-specific incidence of infection was lowest for 3-4-year olds (3.3%) and increased gradually up to the age of 20 24 years (10.8%). The number of notified cases was highest for 3-9-year olds. CONCLUSIONS: In the Dutch population B. pertussis infections occur more frequently and in elder age-categories then suggested by notifications. Mathematical modeling could explore what booster vaccination strategies are most effective in reducing severe disease among not (completely) vaccinated infants. PMID- 16352343 TI - A MEMS fabricated flexible electrode array for recording surface field potentials. AB - We developed a method to microfabricate flexible electrode arrays on a thin Kapton substrate, which was engineered to minimize trauma when inserted between the dura and skull to obtain surface EEG recordings. The array consisted of 64 gold electrodes, each 150microm in diameter on a 750microm spaced 8x8 grid. Using photolithographic procedures, any arrangement of electrodes can be implemented. We used the electrode array to record evoked response signals to create topographical maps of the whisker barrels on the cortical surface with excellent signal stability over a period of 8h. The materials used for this fabrication are potentially biologically inert and, with some additional modifications to the design, can be chronically implanted with minimal side effects. Retinal prosthesis, human neurosurgery, and neurological research are all limited to some degree by the resolution and biological compatibility of the implants used. This type of array could greatly enhance the spatial resolution, signal quality, and stability of implantable surface electrode arrays. PMID- 16352344 TI - Health status, resource consumption, and costs of dysthymia. A multi-center two year longitudinal study. AB - BACKGROUND: In this study we estimated the health status, resource consumption and costs of a large cohort of patients with early and late-onset dysthymia. METHODS: The DYSCO (DYSthymia COsts) project is a multi-center observational study which prospectively followed for two years a randomly chosen sample of patients with dysthymia in the Italian primary health care system. RESULTS: A total of 501 patients were followed for two years; 81% had early-onset dysthymic disorder. During the study, improvement was seen in most domains of the 36-Item Short Form Health Survey (SF-36) questionnaire. Comparison of the SF-36 scores for the two groups showed that only the physical health index significantly differed during the two years. The use of outpatient consultations, laboratory tests and diagnostic procedures was similar in the two groups, but patients with early-onset dysthymia were admitted significantly more than late-onset cases. Hospital admissions were almost entirely responsible for the higher total cost per patient per year of early-onset dysthymia. LIMITATIONS: A first limitation of this study is that general practitioners were selected on the basis of their willingness to participate, not at random; secondly, no information was collected on concomitant psychiatric comorbidities. CONCLUSIONS: The present study provides the first prospective, long-term data on service use and costs in patients with dysthymia. Differently from patients with early-onset dysthymia, patients with late-onset dysthymia were admitted less and cost less. PMID- 16352345 TI - Assessment of suicidality in a Moroccan metropolitan area. AB - BACKGROUND: The aim of the study was to evaluate the prevalence of suicidal ideations and suicide attempts in a representative sample of the general population of the urban area of Casablanca, Morocco. METHODS: The survey was conducted based on face-to-face household interviews. The Mini International Neuropsychiatric Interview (M.I.N.I.) was used to assess axis I diagnoses according to DSM-IV criteria and the M.I.N.I. suicidality module to rate the severity of active suicidality. RESULTS: The 1-month prevalence of suicidal ideation was 6.3%. Seventeen subjects (2.1%) reported at least one suicide attempt during their lifetime. Some variables were positively associated to suicidal ideation: the non-married status, subjects with a history of psychiatric disorders, and subjects without children. At least one mental disorder was present among 88.2% of subjects with suicidal ideation. Major depressive disorder was the most prevalent one (23.5%). LIMITATIONS: The sample was small and the prevalence was not determined longitudinally. CONCLUSION: Suicidal ideation being relatively frequent in the general population, there is a need to develop programs of prevention of suicide. PMID- 16352346 TI - Major depressive disorder in adolescents: family psychiatric history predicts severe behavioral disinhibition. AB - BACKGROUND: Major Depressive Disorder (MDD) becomes increasingly prevalent during adolescence and is associated with substantial psychiatric comorbidity and psychosocial impairment. The marked behavioral heterogeneity evident among adolescents with MDD suggests the possibility of distinct subtypes. This study was designed to determine whether family psychiatric histories differ between groups of MDD adolescents defined by the presence or absence of severe behavioral disinhibition. METHODS: Adolescents with MDD (n = 71) completed the Buss-Durkee Hostility Inventory--Adapted, Adolescent Aggressive Incidents Interview (AAII), Measure of Aggression, Violence, and Rage in Children, Diagnostic Interview Schedule for Children, Suicidal Ideation Questionnaire-JR., Suicidal Behavior Inventory, and Reynolds Adolescent Depression Scale. Parents completed the Family Informant Schedule and Criteria, Children's Affective Liability Scale, AAII, and a partial DISC. Behavioral disinhibition (BD) measures were used to assign adolescents to MDD+BD (n = 41) and MDD-BD (n = 30) groups. RESULTS: The MDD+BD group had a higher prevalence of drug use disorders in biological fathers than the MDD-BD group. The MDD+BD group also had higher proportions of paternal second degree relatives with alcohol use disorders, drug use disorders, and psychiatric hospitalizations, and a higher proportion of maternal second degree relatives with antisocial personality disorder. LIMITATIONS: Limitations include reliance on single informants for family psychiatric histories and the failure to distinguish between child- and adolescent-onset depression. CONCLUSIONS: Family psychiatric histories differentiated MDD adolescents grouped by the presence or absence of behavioral disinhibition, suggesting possible etiologic mechanisms. Further research on subtypes or comorbid presentations may assist in the development of targeted treatment strategies. PMID- 16352347 TI - Relationships of categorization on tests and daily tasks in patients with schizophrenia, post-stroke patients and healthy controls. AB - The purpose of this study was to explore the relationships between performance on formal categorization-sorting tests and daily tasks that require sorting skills among individuals with schizophrenia compared with post-stroke and healthy control groups. Furthermore, the relationships among the performances of patients with schizophrenia on the various tests were studied. The study included 70 participants: 37 patients with schizophrenia treated at a community day center, 18 post-stroke patients, and 15 healthy controls. Subjects were evaluated on four different categorization tests (Wisconsin Card Sorting Test (WCST), Short Category Test (SCT), Loewenstein Occupational Therapy Cognitive Assessment (LOTCA), and Riska Object Classification (ROC); and five different functional daily tasks that require categorization abilities. Results showed significant differences between the performance of patients and control subjects on both tests and daily tasks. Spearman correlations within the schizophrenic group between test scores showed moderate significant correlations, and between tests and daily tasks showed low to moderate significant correlations. These findings suggest that weaker, yet significant relationships were found between performance on cognitive tests and daily tasks than between cognitive tests tapping the same or similar constructs. This type of difference would be expected; however, further research is needed to clarify the underlying components of categorization tests and their relations to daily task performance of individuals with schizophrenia. PMID- 16352348 TI - Effect of low-frequency transcranial magnetic stimulation on an affective go/no go task in patients with major depression: role of stimulation site and depression severity. AB - Repetitive transcranial magnetic stimulation (rTMS) holds promise as a therapeutic tool in major depression. However, a means to assess the effects of a single rTMS session on mood to guide subsequent sessions would be desirable. The present study examined the effects of a single rTMS session on an affective go/no go task known to measure emotional-cognitive deficits associated with major depression. Ten patients with an acute episode of unipolar major depression and eight partially or completely remitted (improved) patients underwent 1 Hz rTMS over the left and right dorsolateral prefrontal cortex prior to task performance. TMS over the mesial occipital cortex was used as a control. We observed significantly improved performance in depressed patients following right prefrontal rTMS. This beneficial effect declined with decreasing depression severity and tended to reverse in the improved group. Left prefrontal rTMS had no significant effect in the depressed group, but it resulted in impaired task performance in the improved group. Our findings indicate that the acute response of depressed patients to rTMS varies with the stimulation site and depression severity. Further studies are needed to determine whether the present paradigm could be used to predict antidepressant treatment success or to individualize stimulation parameters according to specific pathology. PMID- 16352349 TI - HCMV-encoded G-protein-coupled receptors as constitutively active modulators of cellular signaling networks. AB - Several herpesviruses encode G-protein-coupled receptor (vGPCR) proteins that are homologous to human chemokine receptors. In contrast to chemokine receptors, many vGPCRs signal in a ligand-independent (constitutive) manner. Such constitutive signaling is of major significance because various pathologies are associated with activating GPCR mutations. Constitutive activity of the human herpesvirus 8 encoded GPCR (ORF74), for example, is essential for its oncogenic potential to cause angioproliferative Kaposi's sarcoma-like lesions. The human cytomegalovirus (HCMV) encodes four GPCRs, of which US28 and UL33 display constitutive activity in transfected, but also HCMV-infected, cells. In addition, US28 is activated by a broad spectrum of chemokines. Furthermore, both US28 and UL33 show promiscuous G-protein coupling, whereas chemokine receptors activate primarily G(i/o) proteins. Thus, these vGPCRs are versatile signaling devices, reprogramming cellular signaling networks to modulate cellular function after infection. By these means, these HCMV-encoded receptors might contribute to HCMV-related pathologies. PMID- 16352350 TI - Excision of visual cortex does not affect the facilitatory and inhibitory effects of a light flash prepulse on the acoustic startle reflex in the rat. AB - We studied the inhibitory (PPI) and facilitatory (PPF) behavioural effects of brief light flashes presented as prepulses just prior to the acoustic startle reflex (ASR) in intact rats and in rats that had a bilateral excision of visual cortex (VC). The VC-lesioned and control groups (n=5, 5) were near identical in PPF for flashes presented 5-20 ms before the ASR and near identical in PPI for flashes presented 60 ms before the ASR. These findings differ from those of a case report of a human patient following occipital cortex resection, for whom light flashes failed to produce either acoustic startle PPF or PPI if the flash were presented to the portion of the visual field in which she could not see; they differ also from data obtained in functionally decorticate rats, for which a light flash produced no startle PPI, but instead a late-appearing exaggerated PPF. In the present experiment the lesion was restricted to VC, while in contrast the occipital resection in the human patient included posterior areas of the parietal and temporal lobe, and functional decortication in the rat disengaged the entire cortex. The greater extent of these two effective surgical procedures may have been responsible for their greater behavioural effects; otherwise, the complete loss of reflex control by photic stimulation in the human patient but not in the rat may reflect the greater functional importance and anatomical complexity of the VC in humans. PMID- 16352351 TI - Effects on the function of three trophic levels in marine plankton communities under stress from the antifouling compound zinc pyrithione. AB - This study aimed to investigate functional responses of natural marine planktonic communities to stress from the antifouling compound zinc pyrithione (ZPT). Isotope labelling techniques (14C) were applied to study bacterial incorporation of leucine, photosynthetic activity of phytoplankton and grazing of labelled prey by zooplankton communities for 6 days after exposures to nominal concentrations of 0, 5, 25, 50 nM ZPT in a mesocosm experiment in Isefjord, Denmark. Significant direct effects were visible on chlorophyll a concentrations, which decreased in all exposed communities, to between 48 and 36% of control concentrations on Day 3, 1 day after the last exposure. Phytoplankton activities were also significantly affected on Day 3 with activities between 9 and 26% of control levels, as was zooplankton activities in the 25 and 50 nM exposures. In the 50 nM exposure the total community zooplankton activity was reduced to 25+/-4%, and per individual to 46+/-11% of control levels. Bacterial communities showed positive indirect effects with high activities (up to 183+/-40%) due to higher amounts of available substrate from algal death. Pollution induced community tolerance analyses performed on phytoplankton and bacterial communities at the end of the experiment indicated a development of increased tolerance for phytoplankton in the 50 nM exposed communities, whereas there were no changes in tolerance in the bacterial communities. Multivariate analysis of the integrated functional response by the plankton communities revealed a significant difference (p<0.05) between exposed communities compared to controls in the first 3 days after last exposure and in the end of the experiment. The study provides evidence of diverse effects on the functions of marine plankton communities under stress from a pollutant. Direct effects lead to cascading indirect effects throughout the community, eventually causing different developments. Continuous exposure to ZPT could lead to severe long-term effects, causing more permanent changes in structure and function than observed here. The study demonstrates that it is possible to assess the functional effects of a stressor in a complex mesocosm system, and to determine effects in a complex plankton community, which were not predictable from laboratory studies. PMID- 16352352 TI - Short-term exposure to low concentrations of the synthetic androgen methyltestosterone affects vitellogenin and steroid levels in adult male zebrafish (Danio rerio). AB - Short-term effects of methyltestosterone (MT) on the endocrine system of adult male zebrafish (Danio rerio) were examined. Males were exposed to 0, 4.5, 6.6, 8.5, 19.8, 35.9, 62.3 ng MT/l and ethinylestradiol (EE2) (26.4 ng/l) for 7 days. Several physiological endpoints that may be affected by endocrine disrupters were analysed, specifically vitellogenin (VTG) concentration, estradiol (E2), testosterone (T), and 11-ketotestosterone (KT) content, brain aromatase activity and gene expression of CYP19A1 and CYP19A2 in the testis. Exposure to the lowest MT concentration (4.5 ng MT/l), and the EE2 increased the concentration of VTG significantly compared to solvent control group. Exposure to higher concentrations of MT did not increase VTG levels. Endogenous KT and T levels decreased significantly in a concentration-dependent manner in response to the MT exposure and the lowest effective concentrations were 6.4 and 8.5 ng MT/l, respectively. The levels of KT and T were also significantly suppressed by EE2 when compared to the solvent control group. Significant decreases in endogenous E2 levels were found in some MT groups but it was not possible to distinguish a simple concentration-response relationship. No effects of MT or EE2 on the brain aromatase activity or on testicular gene expression of CYP19A1 and CYP19A2 were detected. The results show that androgens such as MT can act as endocrine disrupters even at very low concentrations. PMID- 16352353 TI - Prevalence of variety of cardiac tumors presenting to tertiary care center in Eastern Mediterranean area over 21 years. PMID- 16352354 TI - Biplane assessment of left ventricular function during atrial fibrillation at beats with equal subsequent cycles. AB - BACKGROUND: Prior study has demonstrated that the biplane single-beat method could be used to assess left ventricular function during atrial fibrillation at a beat with equal subsequent cycles. The study was to test whether we could improve the method by measuring a few beats with equal subsequent cycles and cycle-length limits. METHODS: In 75 patients with atrial fibrillation, stroke volume and ejection fraction were determined from simultaneous biplane views of left ventricle for 20 beats using a matrix-array transducer and a biplane Simpson's rule. The influence of cycle lengths on the values of systolic parameters at beats with equal subsequent cycles was examined from the plot of normalized parameters (measured values/average values) against cycle lengths. The values of 1 to 3 beats with equal subsequent cycles and cycle-length limits were averaged and compared with the average values over 20 beats by Bland-Altman and mean percentage difference analysis. The variability of repeat measurements was evaluated in 10 patients. RESULTS: The systolic parameters measured at beats with cycle lengths shorter than 500 ms were usually far below the average values. Agreement and mean percentage difference analysis revealed improved accuracy when 2 or 3 beats with cycle-length limits (>500 ms) were used for assessment. As the variability of averaging 2 or 3 beats is no greater than that of repeat measurements, both methods are equally good. CONCLUSIONS: Accurate assessment of left ventricular systolic function in atrial fibrillation can be obtained by averaging 2 beats with equal subsequent cycles and cycle-length limits (>500 ms). PMID- 16352355 TI - The deteriorating cardiovascular health in China in the past decade. PMID- 16352356 TI - Cocaine-induced acute myocardial infarction in young individuals with otherwise normal coronary risk profile: is coronary microvascular dysfunction one of the underlying mechanisms? PMID- 16352357 TI - Obesity-related increased perioperative morbidity in CABG patients: does metabolic syndrome affect the perioperative outcomes? PMID- 16352358 TI - Usefulness of neopterin levels and left ventricular function for risk assessment in survivors of acute myocardial infarction. AB - BACKGROUND: Results of previous studies have shown that serum neopterin levels are higher in patients with acute myocardial infarction (AMI). However, little is known regarding the prognostic value of neopterin in survivors of AMI. The present study analyzes the prognostic value of neopterin serum concentration and left ventricular ejection fraction in survivors of AMI. METHODS: The additional prognostic information provided by neopterin to parameters of left ventricular function in survivors of AMI was investigated through 82 patients with ST-segment elevation. RESULTS: Binary logistic regression analysis identified the following predictors of 6-month cardiac death: Killip class >I at presentation (OR=2.4, 95% CI: 1.2 to 3.8, p=0.001), a reduced ejection fraction (OR=1.9, 95% CI: 1.3 to 2.2, p=0.001), and greater neopterin (OR=1.4, 95% CI: 1.1 to 1.6, p=0.01). CONCLUSIONS: Neopterin is associated with mortality, in addition to parameters of ventricular function (Killip class and ejection fraction) in survivors of AMI. PMID- 16352359 TI - Optimal quality reporting in markets for health plans. AB - Quality reports about health plans and providers are becoming more prevalent in health care markets. This paper casts the decision about what information to report to consumers about health plans as a policy decision. In a market with adverse selection, complete information about quality leads to inefficient outcomes. In a Rothschild-Stiglitz model, we show that averaging quality information into a summary report can enforce pooling in health insurance, and by choice of the right weights in the averaged report, a payer or regulator can induce first-best quality choices. The optimal quality report is as powerful as optimal risk adjustment in correcting adverse selection inefficiencies. PMID- 16352360 TI - The effect of physician-hospital affiliations on hospital prices in California. AB - During the 1990s, a record number of U.S. hospitals entered into some form of vertical combination with physicians. During the same period, many integrated hospital-physician arrangements broke up. Using data from California, we investigate whether such vertical activity affected hospital pricing. We find that neither integration nor disintegration was associated with significant changes in prices. Integration among rural hospitals is associated with large price decreases, but the sample of such hospitals is small. PMID- 16352361 TI - Phenol removal using Brassica juncea hairy roots: role of inherent peroxidase and H(2)O(2). AB - Removal of phenol, a major pollutant in aqueous effluents was studied using plant hairy root cultures. Among four different species of hairy roots tested, Brassica juncea showed the highest potential for phenol removal. The effect of phenol concentration and reuse in a batch system was studied using B. juncea hairy root cultures. Unlike most of the studies reported earlier, phenol removal by the hairy roots was seen to take place without the need for addition of external hydrogen peroxide (H(2)O(2)). To understand the mechanism of phenol removal, levels of peroxidase and phenol oxidase were monitored in the hairy roots. Peroxidase activity in the roots was enhanced when exposed to phenol, while phenol oxidase remained constant. Since peroxidase has a pre-requisite for H(2)O(2), the levels of H(2)O(2) were monitored for its in situ synthesis. H(2)O(2) levels were seen to increase in the presence of phenol. Thus, a mechanism wherein hairy roots also produce H(2)O(2) besides peroxidase, as a protection strategy of plant against xenobiotic stress is plausible. PMID- 16352362 TI - Treatment of chronic hepatitis C in HIV-positive individuals: selection of candidates. AB - Recent evidence suggests that when HCV therapy is administered adequately (full doses of ribavirin, satisfactory drug compliance, and for at least 12 months irrespective of the HCV genotype) and to the appropriate co-infected candidates, treatment responses may be similar to those seen in HCV mono-infected individuals. The best responders are co-infected individuals under 40 years old with HCV genotypes 2 or 3, low HCV viral load, no cirrhosis, elevated ALT levels, elevated CD4 counts, and low or undetectable plasma HIV-RNA. Treatment should be considered in antiretroviral-naive co-infected patients with stable HIV infection. In patients already on antiretroviral therapy, HCV therapy should only be administered after replacing ddI by another antiretroviral drug. In patients with evidence of advanced liver fibrosis, HCV therapy should be considered as a priority. However, patients with decompensated cirrhosis should not be treated. In patients with CD4 counts < 200 cells/microl and/or plasma HIV-RNA above 100,000 copies/ml, it may be better to consider suppression of HIV replication before beginning HCV therapy. Individuals with a history of severe neuropsychiatric disorders, people who consume a lot of alcohol and those addicted to illegal drugs generally should not be considered suitable for HCV treatment, and efforts should be concentrated on detoxification programmes. PMID- 16352363 TI - Epidemiology of viral hepatitis and HIV co-infection. AB - Worldwide, hepatitis B virus (HBV) accounts for an estimated 370 million chronic infections, hepatitis C virus (HCV) for an estimated 130 million, and HIV for an estimated 40 million. In HIV-infected persons, an estimated 2-4 million have chronic HBV co-infection and 4-5 million have HCV co-infection. HBV, HCV and HIV share common routes of transmission, but they differ in their prevalence by geographic region and the efficiency by which certain types of exposures transmit them. Among HIV-positive persons studied from Western Europe and the USA, chronic HBV infection has been found in 6-14% overall, including 4-6% of heterosexuals, 9 17% of men who have sex with men (MSM), and 7-10% of injection drug users. HCV infection has been found in 25-30% of HIV-positive persons overall; 72-95% of injection drug users, 1-12% of MSM and 9-27% of heterosexuals. The characteristics of HIV infected persons differ according to the co-infecting hepatitis virus, their epidemiologic patterns may change over time, and surveillance systems are needed to monitor their infection patterns in order to ensure that prevention measures are targeted appropriately. PMID- 16352364 TI - Assessment of liver fibrosis in co-infected patients. AB - The evaluation of liver injury in HIV patients co-infected with HBV and HCV should follow the same principles as the evaluation of any patient with chronic liver disease. The initial clinical evaluation should include documentation of risk factors for progressive disease. HIV history is important particularly with respect to a past history of significant or prolonged immunosuppression as this has been clinically correlated with more advanced liver disease. Liver transaminases are an important predictor of disease severity and progression in HIV patients. Liver biopsy has remained the 'gold standard' for the grading of inflammation and staging of disease. We would still recommend liver biopsy in HIV patients particularly those with HCV because recent community-based studies in the HAART era have suggested slower rates of progression for HIV/HCV than studies from tertiary care centres and older cohorts. Since, liver biopsy is invasive and expensive, non-invasive techniques including serological tests and novel imaging techniques have evolved to stage liver fibrosis. A novel technique for measuring hepatic elasticity has recently been validated alone and in combination with serum markers for HCV mono-infection. Future trends for staging liver disease must not only focus on cross sectional diagnosis but on utilizing novel techniques to stratify risk for disease progression over time. PMID- 16352365 TI - Assessment of treatment efficacy in HBV infection and disease. AB - In patients co-infected with HIV and HBV viral load monitoring allows comparison of the efficacy of different antiviral drugs and determination of the initial response to therapy. Usually, the decline in viral load is associated with an improvement in ALT levels and liver histology scores, but some patients show transient elevations in ALT levels due to the restoration of the specific anti HBV immune response. Analysis of liver histology is useful to determine the improvement of the inflammatory activity of the disease and liver fibrosis, and to make an accurate diagnosis when drug hepatotoxicity is suspected. HBe seroconversion and HBs loss have been described in some patients during anti-HBV therapy with lamivudine, adefovir or tenofovir, indicating that the control of viral replication by these agents may lead to a dramatic suppression of viral load. HBV drug resistant mutants can be diagnosed at an early stage with the monitoring of viral load. Future research directions include the determination of predictive factors of response, the effect of anti-HBV therapy on liver disease severity, the impact of long-term treatment on HBsAg clearance and intrahepatic cccDNA, the impact of HBV drug resistance on the liver disease, the role of cross resistance testing of HBV strains in patients with HBV treatment failure. PMID- 16352366 TI - Management of end stage liver disease (ESLD): what is the current role of orthotopic liver transplantation (OLT)? AB - Liver disease due to chronic hepatitis B and C is now a leading cause of morbidity and mortality among HIV-infected patients in the developed world, where classical opportunistic complications of severe immunodeficiency have declined dramatically. Orthotopic liver transplantation (OLT) is the only therapeutic option for patients with end-stage liver disease (ESLD). Accumulated experience in North America and Europe in the last 5 years indicates that 3-year survival in selected HIV-infected recipients of liver transplants was similar to that of HIV negative recipients. So, HIV infection by itself is not therefore a contraindication for liver transplantation. As survival of HIV-infected patients with ESLD is shorter than non-HIV-infected population, the evaluation for OLT should be made after the first liver decompensation. The current selection criteria for HIV-positive transplant candidates include: no history of opportunistic infections or HIV-related neoplasms, CD4 cell count > 100 cells/mm(3), and plasma HIV viral load suppressible with antiretroviral treatment. For drug abusers, a 2-year abstinence from heroin and cocaine is required, although patients can be in a methadone programme. The main problems in the post-transplant period are pharmacokinetic and pharmacodynamic interactions between antiretrovirals and immunosuppressive drugs, and the management of relapse of HCV infection. Up to now, experience with pegylated interferon and ribavirin is scarce in this population. Currently, HCV re-infection is the main cause for concern. PMID- 16352367 TI - Prevention of viral hepatitis in HIV co-infection. AB - Co-infection of HIV-positive patients with hepatitis viruses worsens the long term prognosis and this is summative for each new infection in any individual. Vaccination against hepatitis A or B may be effective but response rates are reduced in HIV infected patients. Improvement in response can be induced through extra doses, higher doses and HAART-induced increase in the CD4 count. Hepatitis B and C may also be prevented through counselling about safer sex, particularly condom use. In intravenous drug users, harm reduction, counselling and the use of needle/syringe exchange schemes may be helpful. PMID- 16352368 TI - Treatment algorithm for chronic hepatitis B in HIV-infected patients. AB - Recommendations for the treatment of chronic hepatitis B (CHB) in HIV-infected patients is complex due to the lack of controlled trials and the dual activity of therapeutic agents on both viruses. Thus, proposals for optimal anti-HBV therapy in HIV-infected patients should be pragmatic using the knowledge from HBV mono- and HIV/HBV co-infected studies. There are four approved drugs for the treatment of CHB which include interferon alpha (IFN), lamivudine (LAM), entecavir (ETV) and adefovir dipivoxil (ADV). LAM, tenofovir disoproxil fumarate (TDF) and emtricitabine (FTC) are approved for HIV and active against HBV. Studies with IFN are limited in HIV/HBV co-infected patients but suggest a decreased response compared with HBV mono-infected patients. LAM and FTC are effective against HBV but are associated with a high rate of HBV resistance. ETV, ADV and TDF are effective against wild-type and LAM-resistant HBV with a favourable resistance profile shown for ADV and TDF. Interferon, ADV or ETV are the preferable drugs in HBV naive patients who do not require HIV therapy. Combination of TDF plus FTC or LAM should be proposed in patients with therapeutic indication for both viruses. TDF should be included in the anti-retroviral regiment of patients with HBV resistance to lamivudine. PMID- 16352369 TI - Multiple viral infections. AB - Individuals at risk of HIV are concomitantly at risk of acquiring parenterally or sexually transmitted viruses. Multiple hepatitis co-infection (HBV+HCV; HBV+HDV; HBV+HDV+HCV) has not been systematically sought after in the large cohorts of HIV infected patients, but has been reported in 0.4% to more than 50% of patients. HIV-infected patients with multiple hepatitis have a higher rate of liver-related morbidity and mortality than patients with HIV infection alone or with a single hepatitis co-infection. The degree of immunodepression is an important factor in liver disease progression. Since GBV-C virus is transmitted parenterally or by sexual contact, a high prevalence was found in chronic hepatitis C and in HIV infected patients. Patients with multiple hepatitis have been excluded from randomised therapeutic trials of viral hepatitis in HIV-infected and HIV-negative patients. Thus, the therapeutic approach is based on the results of a small series and empirically oriented toward the prevailing infection. HIV-infected patients should be tested for hepatitis B, C and D systematically and hepatitis B vaccination should be considered for those with HCV co-infection and absence of HBV markers. Studies are needed to assess treatment strategies. PMID- 16352370 TI - Acceptability of community and health facility-based directly observed treatment of tuberculosis in Tanzanian urban setting. AB - SETTING: An urban district in Dar es Salaam, Tanzania. OBJECTIVE: To assess the acceptability of community and health facility-based direct observation of treatment (DOT) of tuberculosis (TB) patients in Temeke district. DESIGN: Both quantitative and qualitative study methodologies were used. Eight focus group discussions were carried out with TB patients, treatment supporters, health workers and community members. Quantitative study was also carried out among TB patients and treatment supporters. RESULTS: A total of 268 patients and 103 treatment supporters were enrolled in the study. The majority of the patients (75%) were satisfied with the DOT options they received. Males were more satisfied with community-based DOT (84%) than females (67%) (OR 4.96, 95% CI 1.38 17.86). The majority (81%) were willing to supervise another TB patient. The results of the qualitative study demonstrated that community-based DOT was preferred because it was found to be convenient, reduced costs, saved time of the patients and reduced workload in health services. The main challenge of community based DOT was to ensure effective supervision and monitoring of patients and treatment supporters in the community. CONCLUSION: The results from the quantitative and qualitative studies indicate that both DOT options were acceptable. The study has also identified key challenges and opportunities for effective implementation of community-based DOT interventions that are relevant, sensitive and acceptable to the population. Community-based DOT is a viable option and can complement and strengthen the existing health facility-based DOT, especially in countries like Tanzania where the health system is overwhelmed with increasing number of TB and HIV/AIDS patients. PMID- 16352371 TI - Developing a medicine education program in Finland: lessons learned. AB - The aim of this article is to describe the process of developing a medicine education program for elementary and middle schools in Finland and the lessons learned during the process. Further described is how teachers evaluated the usefulness of the medicine education materials created during the process. By medicine education we mean education about the proper use of medicines, abuse of medicines being just a small part of it. The development process started in 2002 by conducting focus group discussions with children in order to discover how children of different ages understand medicine-related topics. Moreover, teachers completed questionnaires in 2002 to assess their opinions about the importance of medicine education as a part of school health education. Based on the results of these two studies, materials were created during 2002-2003 (, in Finnish with an English introduction). These materials gave the teachers information about the proper use of medicines and some ideas for assignments. As a last part of this research project in autumn 2003, the materials were piloted by a group of elementary and middle school teachers (n=14), and the usefulness of the materials were evaluated during focus group discussions after a teaching period. Based on the evaluation, we learned that the Website should contain a simple structure and ready-to-use materials in order to be used by teachers. Moreover, the fact that teachers need information in order to be able to teach this unfamiliar topic became clear. Teachers of younger children need concise information, but teachers of adolescents need more in-depth information. Furthermore, teachers may have negative attitudes towards medicines, and therefore, medicine education should be rationalized for them. We conclude our article with recommendations on what should be taken into consideration when medicine education programs are planned. PMID- 16352372 TI - A case of recurrent non-small-cell lung carcinoma and paraneoplastic Cushing's syndrome. AB - Secretion of ectopic adrenocorticotropic hormone (ACTH) with consequently Cushing's syndrome is a rare paraneoplastic phenomenon. It has been described in a variety of malignancies, like bronchial carcinoids, small-cell lung carcinoma, thymoma, pancreatic carcinoma and other. In many cases of suspected ectopic ACTH secretion, it is difficult to histologically or cytochemically confirm the diagnosis. We present a 63-year-old woman with a recurrent poorly differentiated squamous cell lung carcinoma with clinical and biochemical features consistent with ectopic Cushing's syndrome. Immunocytochemical staining confirmed the secretion of ACTH by tumour cells. PMID- 16352373 TI - Oleate accumulation, induced by silencing of microsomal omega-6 desaturase, declines with leaf expansion in transgenic tobacco. AB - All higher plants contain at least one microsomal omega-6 desaturase (FAD2), which inserts a double bond between the carbons 12 and 13 of monounsaturated oleic acid to generate polyunsaturated linoleic acid and controls most of the polyunsaturated lipid synthesis in plant cells. RNA interference can be used to silence endogenous genes by effective degradation of target transcripts. To investigate development-related silencing of the FAD2, fatty acid composition was analyzed in the context of leaf expansion in FAD2-silenced tobacco lines obtained by RNA interference technology. We observed that the increased oleate level in unexpanded leaves due to FAD2-silencing receded significantly in fully expanded leaves. The mechanism involved in this interesting phenomenon was investigated by analyses of individual lipid proportion, fatty acid composition of individual lipids, and FAD2 transcript level in the transgenic leaves at different expansion stages. Data revealed that the expansion-related FAD2-silencing effect was not due to rebound of FAD2 transcript, but rather probably due to chloroplast development with leaf expansion. PMID- 16352374 TI - Breast-feeding, maternal feeding practices and mealtime negativity at one year. AB - This paper explores whether breast-feeding, mediated by lower maternal use of controlling strategies, predicts more positive mealtime interactions between mothers and their 1 year old infants. Eighty-seven women completed questionnaires regarding breast-feeding, assessing their control over child feeding and mealtime negativity at 1 year of infant age. Seventy-four of these women were also observed feeding their infants solid food at 1 year. Mediation analyses demonstrated that the experience of breast-feeding, mediated by lower reported maternal control over child feeding, predicted maternal reports of less negative mealtime interactions. The experience of breast-feeding also predicted observations of less conflict at mealtimes, mediated by observations of maternal sensitivity during feeding interactions. The implications of these findings are discussed. PMID- 16352375 TI - Differential regulation of proteasome activity in the nucleus and the synaptic terminals. AB - Proteasome is a multi-subunit proteolytic complex that degrades proteins covalently linked to multiple molecules of ubiquitin. Earlier studies showed a role for the ubiquitin-proteasome pathway in several models of long-term memory and other forms of synaptic plasticity. In Aplysia, the ubiquitin-proteasome pathway has been shown to contribute to the induction of long-term facilitation. In other model systems, ubiquitin-proteasome-mediated proteolysis has also been shown to play a role in synapse development. Previous studies of synaptic plasticity focused on changes in components or the substrates of the ubiquitin proteasome pathway in whole neurons. Modification of specific synapses would require precise spatial and temporal regulation of the components of the ubiquitin-proteasome pathway within the subcellular compartments of neurons during learning. As a first step towards testing the idea of local regulation of the ubiquitin-proteasome pathway in neurons, we investigated proteasome activity in nuclear and synaptosomal fractions. Here we show that proteasome activity in the synaptic terminals is higher compared to the activity in the nucleus in the Aplysia nervous system as well as in the mouse brain. Furthermore, the proteasome activity in the two neuronal compartments is differentially modulated by protein kinases. Differential regulation of proteasome activity in neuronal compartments such as the synaptic terminals is likely to be a key mechanism underlying synapse specific plasticity. PMID- 16352376 TI - The burden of Herpes Zoster: a prospective population based study. AB - We analysed prospectively the medical, societal and economic burden among patients from 18 general practices in East London, serving 158,716 patients who presented to their general practitioners with acute Herpes Zoster over an 8-month period. One hundred and eighty-six patients with HZ were seen by GPs during the study period, of whom 96 were referred, 70 enrolled and 65 completed. PHN occurred in 13.4% of patients. The average overall cost of HZ in the first 6 months was calculated at pound524 per patient. Medical costs were highest in those aged over 65 and societal costs highest in those aged under 65 years. PMID- 16352377 TI - Immunization with dendritic cells retrovirally transduced with mycobacterial antigen 85A gene elicits the specific cellular immunity including cytotoxic T lymphocyte activity specific to an epitope on antigen 85A. AB - In the present study, we evaluated antigen 85A (Ag85A) gene-transduced dendritic cells (DCs) vaccine against Mycobacterium tuberculosis. Murine bone marrow derived DCs were retrovirally transduced with mycobacterial Ag85A gene and injected to BALB/c mice intravenously. The DC vaccine was capable of inducing purified protein derivative (PPD)- and the antigen-specific spleen cell proliferation and IFN-gamma production from both CD4+ and CD8+ T cells in spleens of the immune mice. In addition, the DC vaccination induced cytotoxic T lymphocytes (CTL) and IFN-gamma-producing cells specific for a 9-mer CTL epitope on Ag85A molecule. This eliciting cellular immunity led to protection against wasting disease due to M. tuberculosis infection and induction of moderate bacterial clearance. PMID- 16352378 TI - Re: Perception by the public and medical profession of oral and maxillofacial surgery--has it changed after 10 years? PMID- 16352379 TI - Improved sensory relearning after nerve repair induced by selective temporary anaesthesia - a new concept in hand rehabilitation. AB - The outcome after nerve repair in adults is generally poor. We hypothesized that forearm deafferentation would enhance the sensory outcome by increasing the cortical hand representation. A prospective, randomized, double-blind study was designed to investigate the effects of cutaneous forearm anaesthesia combined with sensory re-education on the outcome after ulnar or median nerve repair. During a 2 week period, a local anaesthetic cream (EMLA (n = 7) or placebo (n=6) was applied repeatedly onto the flexor aspect of the forearm of the injured arm and combined with sensory re-education. Evaluation of sensory function was carried out at regular intervals and at 4 weeks after the last EMLA/placebo session. The EMLA group showed significant improvement compared to placebo in perception of touch/pressure, tactile gnosis and in the summarized outcome after 6 weeks. These results suggest that cutaneous forearm anaesthesia of the injured limb, in combination with sensory re-education, can enhance sensory recovery after nerve repair. PMID- 16352380 TI - A spatially-evaluated methodology for assessing risk to a population from contaminated land. AB - A methodology is proposed which combines quantitative probabilistic human health risk assessment and spatial statistical methods (geostatistics) to produce an assessment of risks to human health from exposure to contaminated land, in a manner which preserves the spatial distribution of risks and provides a measure of uncertainty in the assessment. Maps of soil contaminant levels, which incorporate uncertainty, are produced from sparse sample data using sequential indicator simulation. A real, age-stratified population is mapped across the contaminated area, and intake of soil contaminants by individuals is calculated probabilistically using an adaptation of the Contaminated Land Exposure Assessment (CLEA) model. An abundance of information is contained in results which can be interrogated at the population and individual level, and mapped to provide a powerful visual tool for risk managers, enabling efficient targeting of risk reduction measures to different locations. PMID- 16352382 TI - Relapse to smoking. AB - Relapse is by far the most likely outcome of any smoking cessation attempt, even those made with the benefit intensive psychosocial treatment and pharmacotherapy. The present article briefly reviews the epidemiology of smoking and self quitting, the outcome data for major forms of behavioral and pharmacologic smoking cessation treatments, and what is known about the natural history of relapse and recovery among treated smokers. A recent trend in smoking relapse research has been to study the dynamics of key motivational processes, such as withdrawal symptoms, negative affect, and craving, in the laboratory and in smokers' natural environments. This literature is also briefly reviewed, with an emphasis on how such investigations may reveal the limitations of current cessation treatments. Finally, three significant research themes that are likely to be important in future relapse research are highlighted--the possible "hardening" of the smoking population, the potential for developmental research to deepen our understanding of smoking motivation, and the promise of molecular genetic studies for advancing treatment and our understanding of relapse. PMID- 16352383 TI - An analysis of genetic toxicity, reproductive and developmental toxicity, and carcinogenicity data: II. Identification of genotoxicants, reprotoxicants, and carcinogens using in silico methods. AB - This study examined a novel method to identify carcinogens that employed expanded data sets composed of in silico data pooled with actual experimental genetic toxicity (genetox) and reproductive and developmental toxicity (reprotox) data. We constructed 21 modules using the MC4PC program including 13 of 14 (11 genetox and 3 reprotox) tests that we found correlated with results of rodent carcinogenicity bioassays (rcbioassays) [Matthews, E.J., Kruhlak, N.L., Cimino, M.C., Benz, R.D., Contrera, J.F., 2005b. An analysis of genetic toxicity, reproductive and developmental toxicity, and carcinogenicity data: I. Identification of carcinogens using surrogate endpoints. Regul. Toxicol. Pharmacol.]. Each of the 21 modules was evaluated by cross-validation experiments and those with high specificity (SP) and positive predictivity (PPV) were used to predict activities of the 1442 chemicals tested for carcinogenicity for which actual genetox or reprotox data were missing. The expanded data sets had approximately 70% in silico data pooled with approximately 30% experimental data. Based upon SP and PPV, the expanded data sets showed good correlation with carcinogenicity testing results and had correlation indicator (CI, the average of SP and PPV) values of 75.5-88.7%. Conversely, expanded data sets for 9 non correlated test endpoints were shown not to correlate with carcinogenicity results (CI values <75%). Results also showed that when Salmonella mutagenic carcinogens were removed from the 12 correlated, expanded data sets, only 7 endpoints showed added value by detecting significantly more additional carcinogens than non-carcinogens. PMID- 16352384 TI - The timing of marriage and childbearing among rural families in Bangladesh: choosing between competing risks. AB - Early marriage and childbearing among girls is often associated with a wide range of negative social and health consequences for young mothers and their infants, and contributes to rapid population growth. This paper presents findings from qualitative research in three villages of rural Bangladesh, where a range of interventions have been promoted to encourage later marriage and childbearing. Data from in-depth interviews and group discussions are used to describe socio cultural supports for early marriage and childbearing, to examine evidence that change towards later marriage and childbearing is beginning, and to analyze the social dynamics behind these change processes. The findings suggest that norms supporting early marriage and childbearing are beginning to erode, and that changing gender ideals and aspirations for women are a key factor in this erosion. Interviews among the poorest families, however, show that this group tends to experience this changing social environment in terms of heightened risks. Marital strategies among the poorest are, above all, strategies for economic survival, and poor families tend to see the costs of education and delayed marriage for daughters as high and the outcomes as uncertain. At the same time, they have also become aware that early marriage and childbearing entails costs and risks. The authors conclude that further targeting of interventions to the poorest families may help to influence the economic strategies that so often result in early marriage. PMID- 16352385 TI - Culturally compelling strategies for behaviour change: a social ecology model and case study in malaria prevention. AB - Behaviour change is notoriously difficult to initiate and sustain, and the reasons why efforts to promote healthy behaviours fail are coming under increasing scrutiny. To be successful, health interventions should build on existing practices, skills and priorities, recognise the constraints on human behaviour, and either feature community mobilisation or target those most receptive to change. Furthermore, interventions should strive to be culturally compelling, not merely culturally appropriate: they must engage local communities and nestle within social and ecological landscapes. In this paper, we propose a social ecology perspective to make explicit the links between intention to change, actual behaviour change, and subsequent health impact, as relating to both theory-based models and practical strategies for triggering behaviour change. A social ecology model focuses attention on the contexts of behaviour when designing, implementing or critically evaluating interventions. As a case study, we reflect on a community-directed intervention in rural Gambia designed to reduce malaria by promoting a relatively simple and low-cost behaviour: repairing holes in mosquito bednets. In phase 1, contextual information on bednet usage, transactions and repairs (the 'social lives' of nets) was documented. In phase 2 (intervention), songs were composed and posters displayed by community members to encourage repairs, creating a sense of ownership and a compelling medium for the transmission of health messages. In phase 3 (evaluation), qualitative and quantitative data showed that household responses were particularly rapid and extensive, with significant increase in bednet repairs (p<0.001), despite considerable constraints on human agency. We highlight a promising approach-using songs-as a vehicle for change, and present a framework to embed the design, implementation and critical evaluation of interventions within the larger context-or social ecology-of behaviour practices that are the bedrock of health interventions. PMID- 16352386 TI - Feasibility of conducting human studies to address bromate risks. AB - Findings from epidemiologic studies have been important in evaluating risk of exposure to many contaminants in drinking water. In the case of bromate, a byproduct of ozone disinfection of water, it is unlikely that observational studies of populations exposed to bromate in drinking water will be as revealing as studies of other contaminants, unless risks are much higher than predicted from laboratory studies of rodents. Occupational exposure to bromate has occurred in the flour milling and baking industries, as well as in chemical production of potassium bromate, used as a flour additive. The feasibility of a cohort study of bromate-exposed workers should be evaluated by studying the conditions and levels of exposure in these occupational settings. Bromate exposure causes oxidative damage to guanine bases of DNA, producing 8-hydroxy-guanine (8-OH-Gua), which is excised by 8-oxoguanosine glycosylase (OGG1) and excreted in the urine. Polymorphic variants of OGG1 in human populations have been associated with elevated cancer risk. 8-OH-Gua and 8-hydroxy-deoxyguanosine (8-OHdG) have been used as biomarkers of oxidative damage in many human studies, and it would be feasible to employ these indicators in controlled clinical experimental settings to see if exposure to bromate in water at levels close to the maximum contaminant level influences urinary levels of excretion, and if so, to help quantify the level of oxidative damage. Such a study could fill an important data gap by providing human data to help estimate the carcinogenic risk from this exposure. PMID- 16352387 TI - Low-dose exposure of intestinal epithelial cells to formaldehyde results in MAP kinase activation and molecular alteration of the focal adhesion protein paxillin. AB - We investigated the potential pathophysiological role of non-lethal formaldehyde concentrations on human intestinal epithelial HT-29 cells. Expression levels of actin, tubulin and detectable cytokeratin isoforms 5, 13, 18, 19 and 20 were not affected after 24h of exposure to 1mM formaldehyde. By contrast, cellular organization of cytoskeletal constituents was already changed after 60 min. Within 15 min, formaldehyde induced profound tyrosine phosphorylation of the focal adhesion protein paxillin and of proteins at about 120-130 kDa. Concomitantly, phosphorylation of ERK-1/2 and p38 MAP kinase occurred. Paxillin was not only tyrosine phosphorylated but underwent a sustained molecular weight shift representing serine/threonine phosphorylation that was independent of MAP kinase activity and EGF-R-mediated signalling. Our data show that exposure of intestinal epithelial cells to low-dose formaldehyde is followed by rapid and profound signalling events. The data suggest a modifier role of environmental or endogenous formaldehyde for epithelial cell functions. PMID- 16352388 TI - Strange feelings: do amygdala abnormalities dysregulate the emotional brain in schizophrenia? AB - Schizophrenia is widely regarded to be a neurocognitive disorder, i.e. a dysfunction of the neural and cognitive systems subserving thinking and reasoning, memory, language, attention and perception. However, although cognitive dysfunction is certainly a cardinal feature of schizophrenia, we argue that dysfunction of emotional brain systems may be even more important in understanding the disorder. Indeed, in recent years research on the emotional aspects of schizophrenia is accumulating at a high rate. Here, we review the available evidence regarding behavioral and neural manifestations of abnormal emotional systems in schizophrenia. This evidence comes from patient studies using tasks of emotion recognition, emotional expression and emotional experience. Furthermore, studies of schizophrenia patients using structural MRI have demonstrated volume reductions of the amygdala, a key structure of the emotional brain. Finally, functional fMRI studies have revealed an attenuated response of the amygdala to emotional stimuli as compared to neutral stimuli. Beyond demonstrating that dysfunction of the emotional brain is a hallmark of schizophrenia, we propose a model that integrates previous neural accounts of emotional abnormalities in schizophrenia, and specifies a neural basis for differential emotional correlates of positive and negative symptoms. Specifically, a lesion to the amygdala in combination with reduced interconnectivity with the prefrontal cortex is hypothesized to give rise to reduced emotional expression (affective flattening) and emotion recognition deficits. In contrast, an imbalance in dopamine systems may underlie increased anxiety and autonomic arousal, and the assignment of emotional salience to insignificant stimuli, associated with psychosis. We also hypothesize that the central and basolateral nuclei of the amygdala may contribute differentially to these abnormalities. PMID- 16352389 TI - Combination of calcium ionophore A23187 with puromycin salvages human unfertilized oocytes after ICSI. AB - OBJECTIVE: To determine whether oocyte activation using a combination of calcium ionophore A23187 (A23187) with puromycin could salvage human unfertilized oocytes after ICSI. STUDY DESIGN: One hundred and thirteen discarded unfertilized oocytes 20-68 h after ICSI were assigned to four groups: ICSI 20-h group, ICSI 44-h group, ICSI 68-h group and control. All unfertilized oocytes were exposed to A23187 (5 microM) for 5 min and subsequently were incubated with puromycin (10 microg/ml) for 4 h. Sex chromosomal analysis was performed by dual color fluorescence in situ hybridization (FISH). RESULTS: The combination of A23187 with puromycin could activate the unfertilized oocytes 20-68 h after ICSI. The best results were achieved in the ICSI 20-h group, which exhibited an activation rate of 91.2% (31/34), a cleavage rate of 64.7% (22/34) and 44.1% (15/34) high quality embryos. The activation rate, cleavage rate and the number of high quality embryos appeared to decrease with the cultured time of unfertilized oocytes after ICSI. FISH analysis showed six embryos with XX and seven embryos with XY in 16 embryos derived from 2PN2PB. CONCLUSIONS: The combination of calcium ionophore A23187 with puromycin could effectively salvage unfertilized oocytes within 20 h after ICSI. PMID- 16352390 TI - Changes in the treatment of patients with subarachnoid haemorrhage following publication of the International Subarachnoid Aneurysm Trial. PMID- 16352391 TI - Carbonate effects on hexavalent uranium removal from water by nanocrystalline titanium dioxide. AB - A novel nanocrystalline titanium dioxide was used to treat depleted uranium (DU) contaminated water under neutral and alkaline conditions. The novel material had a total surface area of 329 m(2)/g, total surface site density of 11.0 sites/nm(2), total pore volume of 0.415 cm(3)/g and crystallite size of 6.0 nm. It was used in batch tests to remove U(VI) from synthetic solutions and contaminated water. However, the capacity of the nanocrystalline titanium dioxide to remove U(VI) from water decreased in the presence of inorganic carbonate at pH > 6.0. Adsorption isotherms, Fourier transform infrared (FTIR) spectroscopy, and surface charge measurements were used to investigate the causes of the reduced capacity. The surface charge and the FTIR measurements suggested that the adsorbed U(VI) species was not complexed with carbonate at neutral pH values. The decreased capacity of titanium dioxide to remove U(VI) from water in the presence of carbonate at neutral to alkaline pH values was attributed to the aqueous complexation of U(VI) by inorganic carbonate. The nanocrystalline titanium dioxide had four times the capacity of commercially available titanium dixoide (Degussa P-25) to adsorb U(VI) from water at pH 6 and total inorganic carbonate concentration of 0.01 M. Consequently, the novel material was used to treat DU contaminated water at a Department of Defense (DOD) site. PMID- 16352392 TI - The study of various parameters affecting the ion exchange of Cu2+, Zn2+, Ni2+, Cd2+, and Pb2+ from aqueous solution on Dowex 50W synthetic resin. AB - A gel resin containing sulfonate groups (Dowex 50W) was investigated for its sorption properties towards copper, zinc, nickel, cadmium and lead metal ions. The use of selective ion exchange to recover metals from aqueous solution has been studied. The ion exchange behavior of five metals on Dowex 50W, depending on pH, temperature, and contact time and adsorbate amount was studied. Experimental measurements have been made on the batch sorption of toxic metals from aqueous solutions using cation exchanger Dowex 50W. The maximum recoveries (about 97%) Cu(2+), Zn(2+), Ni(2+), Cd(2+) and (about 80%) Pb(2+) were found at pH ranges 8 9. The amount of sorbed metal ion was calculated as 4.1, 4.6, 4.7, 4.8, and 4.7mequiv./gram dry resin for Pb(2+), Cu(2+), Zn(2+), Cd(2+), and Ni(2+), respectively. The precision of the method was examined at under optimum conditions. Selectivity increased in the series: Pb>Cd>Cu>Zn>Ni. It has been observed that, selectivity of the -SO(3)H group of the resin increases with atomic number, valance, degree of ionization of the exchanged metals. The equilibrium ion exchange capacity of resin for metal ions was measured and explored by using Freundlich and Langmuir isotherms. Langmuir type sorption isotherm was suitable for equilibrium studies. PMID- 16352393 TI - A contribution to the debate on LPG fires. PMID- 16352394 TI - Phospholipase C isozymes are differentially distributed in the rat adrenal medulla. AB - Immunohistochemistry has been used to examine the distribution of selected phospholipase C (PLC) isozymes within the adrenal medulla of the rat. PLCbeta isozymes were expressed at moderate levels in the chromaffin cells but more strongly in association with ganglion cell clusters. PLCbeta2 and PLCbeta3 staining of clusters did not overlap suggesting selective PLC isozyme expression in two distinct ganglionic types. The distribution of PLCbeta4 immunoreactivity was very similar to PLCbeta3 with the strongest staining observed in the same cell clusters. Antibodies to PLCbeta1 labelled multiple bands on Western blots and were not therefore used for immunohistochemistry. The chromaffin cells were also immunoreactive for PLCgamma1, although the strongest staining with this antibody was seen in cells surrounding large sinus vessels. PLCdelta1 and PLCdelta2 had quite distinct distributions, with the former selectively localized to an endothelial cell population surrounding the chromaffin cells. This observation was supported by experiments on isolated bovine adrenal medullary cells where PLCdelta1 expression was lost when the cell preparation was enriched for chromaffin cells. Antibodies to PLCdelta2 labelled a network of nerve fibres throughout the medulla and clusters of ganglion cells located primarily at the medullary-cortical boundary. PLCdelta2 immunoreactivity was also present in nerve fibres within the adrenal capsule where it appeared to be co-localized with PLCbeta4 staining. PMID- 16352395 TI - Increased spinal cord NGF levels in rats with cobalamin (vitamin B12) deficiency. AB - We have recently demonstrated that the neuropathological morphological alterations caused by cobalamin (Cbl) deficiency in the rat central nervous system are related to the vitamin's inability to modulate the synthesis of some neurotoxic and neurotrophic agents in opposite directions. In the present study, we measured nerve growth factor (NGF) levels in the spinal cord (SC) and cerebrospinal fluid (CSF) of rats made Cbl-deficient (Cbl-D) by means of total gastrectomy (TG) or a Cbl-D diet. In both cases, Cbl deficiency increased SC and CSF NGF levels after the appearance of myelinolytic lesions in the SC white matter (SCWM) (i.e. after the second post-TG month), and these changes were normalised by Cbl treatment in the 4-month-totally-gastrectomised (TGX) rats. Intracerebroventricular (i.c.v.) anti-NGF-antibody treatment prevented the onset of the myelinolytic SCWM lesions in the 2-month-TGX rats (i.e. when SC and CSF NGF levels are still normal) and normalised the ultrastructure of the SCWM in the 4-month-TGX rats, which was however worsened by the i.c.v. administration of NGF. These findings demonstrate that: (i) Cbl deficiency increases SC and CSF NGF levels; and (ii) endogenous NGF seems to play a noxious role in the progression of rat Cbl-D central neuropathy. PMID- 16352396 TI - Association analysis of the functional Ala111Glu polymorphism of the glyoxalase I gene in panic disorder. AB - The zinc metalloenzyme glyoxalase I (GLO1) is thought to play a role in anxiety disorders because a reduced brain expression of GLO1 has been associated with increased anxiety-behaviours in mice. Recently, a functional Ala111Glu polymorphism in GLO1 has been shown to result in a reduced enzyme activity. The present study tested the hypothesis that this common genetic variant could confer susceptibility to panic disorder using an Italian population sample of 162 panic disorder patients and 288 matched controls. Statistical analysis failed to show association with the overall diagnosis of the disease. However, a weak but significant association was demonstrated between this polymorphism and panic disorder without agoraphobia. While our data suggest that this polymorphism is unlikely to have a major function in the pathogenesis of panic disorder, it could play a role in the subgroup of patients without agoraphobic avoidance. PMID- 16352397 TI - Light delays synaptic deafferentation and potentiates the survival of axotomized retinal ganglion cells. AB - Knowledge of the cellular mechanism underlying the therapeutic effect of stimulation and the optimal doses of such stimulation to maximize neuronal recovery is essential to guide clinical practice in neural rehabilitation. Using hamsters, we transected the optic nerve to demonstrate how light stimulation affects neuronal recovery. The c-fos protein was used as a neuronal connectivity marker. Here we show that: (a) in addition to cell death, a population of cells undergoes synaptic deafferentation and (b) light stimulation delays cell death and deafferentation. Among the three rearing conditions studied (6:18LD, 12:12LD, and 18:6LD), the 12:12LD condition appears to be the one achieving the optimal therapeutic effect. This study provides a solid base in the understanding of the neuroanatomical changes after traumatic brain injury and the need to establish an optimal level and timing for the environmental stimulation. PMID- 16352398 TI - Molecular characterization of two acetylcholinesterase genes from the oriental tobacco budworm, Helicoverpa assulta (Guenee). AB - Acetylcholinesterase (AChE) has been known to be the target of organophosphorous and carbamate insecticides. Only a single AChE, however, existed in insects and was involved in insecticide resistance, recently another AChE is reported in mosquitoes and aphids. We have cloned cDNAs encoding two ace genes, designated as Ha-ace1 and Ha-ace2 by a combined degenerate PCR and RACE strategy from adult heads of the oriental tobacco budworm, Helicoverpa assulta. The Ha-ace1 and Ha ace2 genes encode 664 and 647 amino acids, respectively and have conserved motifs including a catalytic triad, a choline-binding site and an acyl pocket. Both Ha AChEs were determined to be secretory proteins based on the existence of a signal peptide. The Ha-ace1 gene, the first reported ace1 in lepidopterans, belongs to the ace1 subfamily whereas the Ha-ace2 gene showed high similarity to those in the ace2 subfamily. Phylogenetic analysis showed that the Ha-ace1 gene was completely diverged from the Ha-ace2, suggesting that the Ha-ace genes are duplicated. Quantitative real time-PCR revealed that expression level of the Ha ace1 gene was much higher than that of the Ha-ace2 in all body parts examined. The biochemical properties of purified proteins by affinity chromatography showed substrate specificity for acetylthiocholine iodide, and inhibitor specificity for BW284C51 and eserine and their peptide sequences partially identified by a MALDI TOF mass spectrometer demonstrated that two Ha-AChEs were expressed in vivo. PMID- 16352399 TI - Differential cellular distribution of tonicity-induced expression of transcription factor TonEBP in the rat brain following prolonged systemic hypertonicity. AB - In a previous work performed on cerebral cortex and hippocampus we reported that tonicity-responsive enhancer binding protein (TonEBP), originally identified as a transactivator of osmoprotective genes involved in osmoadaptation of renal cells, was induced in neurons only, but to varying levels, following acute systemic hypertonicity. Whether or not this cellular specificity reflected a unique ability of neurons or a differential time course among brain cells for tonicity induction of TonEBP was investigated throughout the brain in this study by subjecting the animals to prolonged systemic hypertonicity. In normal rats, TonEBP immunolabeling and TonEBP-mRNA in situ hybridization labeling showed a widespread, uneven and parallel distribution. TonEBP was expressed primarily in the cell nuclei of neurons, where it was heterogeneously distributed in a nucleoplasmic and a granular pool. In rats subjected to prolonged systemic hypertonicity, TonEBP labeling increased in the cell nuclei of neurons only. The tonicity-induced expression of TonEBP for a given cell group of neurons was rather uniform but varied greatly among neuronal cell groups and was positively correlated with the average size of the cell nuclei, as determined by quantitative analysis of digitized images. The detailed distribution of tonicity induced expression of TonEBP is reported throughout the brain. In normal rats, a very minor proportion of non-neuronal cells, identified as a subset of astrocytes and possibly oligodendrocytes, showed faint nuclear immunolabeling, which however did not increase in hypertonic animals. Ependymocytes, capillary endothelial cells, and microglial cells showed no TonEBP labeling, even in hypertonic animals. Altogether our data indicate that neurons, albeit possibly to a varying extent, are the only brain cells able to use TonEBP-mediated processes for adaptation to a systemic hyperosmotic unbalance. PMID- 16352400 TI - Rhes, the Ras homolog enriched in striatum, is reduced under conditions of dopamine supersensitivity. AB - Striatal dopamine receptors become supersensitive when dopaminergic input is removed through either surgical denervation or pharmacological depletion. Although alterations such as increased D2 receptor binding and increased receptor G protein coupling have been described in supersensitive striatal tissue, their roles in the mechanism of supersensitivity remain uncertain. The Ras Homolog Enriched in Striatum (Rhes) is expressed in brain areas that receive dopaminergic input, and here we test whether alterations in its expression accompany treatments that promote dopamine receptor supersensitivity in rats. Removal of dopamine input to the striatum by surgical denervation with 6-hydroxydopamine resulted in a decrease in rhes mRNA expression throughout striatum, as measured with quantitative in situ hybridization. The decrease was detected as early as two weeks and as late as seven months after surgery. Furthermore, a decrease in rhes mRNA was evident after repeated or acute reserpine treatment. Chronic daily injection of rats with the D2 antagonist eticlopride, which is known to up regulate D2 receptors without inducing profound receptor supersensitivity, did not alter the expression of rhes mRNA in striatum. Thus, changes in rhes mRNA expression are strictly correlated with receptor supersensitivity, perhaps as a result of continuous removal of dopaminergic input. These findings suggest that rhes mRNA expression is maintained by dopamine and may play a role in determining normal dopamine receptor sensitivity. PMID- 16352401 TI - Subcortical middle cerebral artery ischemia abolishes the digit flexion and closing used for grasping in rat skilled reaching. AB - That rats reach for and grasp a food item using a single paw has prompted their use in neurobiological studies of skilled movements and modeling neural injury including middle cerebral artery stroke. Although motor system lesions have been shown to disrupt various qualitative aspects of the transport of a limb to a food target and withdrawal of the limb with the food, no lesion has been found to abolish digit flexion for grasping. Here, rats received unilateral transient middle cerebral artery ischemia that was restricted mainly to subcortical tissue of the forebrain (caudate-putamen, globus pallidus, and associated fibers) or a sham operation. Both paws were later trained and evaluated on skilled reaching using a rating scale for digit use. Middle cerebral artery rats did not flex and close their digits to grasp food when using their contralateral-to-lesion limb. The grasp impairment was not due to a failure to learn the task as middle cerebral artery rats used the ipsilateral limb as successfully as control rats and they were reinforced for reaching by raking food into the reaching box using an open paw. The impairment was also not due to an inability to move the digits, as they were flexed and closed in other phases of the reach. The paradigm should prove useful for further studies of rehabilitation in relation to the idea that digit closure may be controlled by the joint action of a number of neural systems that converge in the basal ganglia. PMID- 16352402 TI - The functional neuroanatomy of working memory: contributions of human brain lesion studies. AB - Studies of patients with focal brain lesions remain critical components of research programs attempting to understand human brain function. Whereas functional imaging typically reveals activity in distributed brain regions that are involved in a task, lesion studies can define which of these brain regions are necessary for a cognitive process. Further, lesion studies are less critical regarding the selection of baseline conditions needed in functional brain imaging research. Lesion studies suggest a functional subdivision of the visuospatial sketchpad of working memory with a ventral stream reaching from occipital to temporal cortex supporting object recognition and a dorsal stream connecting the occipital with parietal cortex enabling spatial operations. The phonological loop can be divided into a phonological short-term store in inferior parietal cortex and an articulatory subvocal rehearsal process relying on brain areas necessary for speech production, i.e. Broca's area, the supplementary motor association area and possibly the cerebellum. More uncertainty exists regarding the role of the prefrontal cortex in working memory. Whereas single cell studies in non-human primates and functional imaging studies in humans have suggested an extension of the ventral and dorsal path into different subregions of the prefrontal cortex, lesion studies together with recent single-cell and imaging studies point to a non-mnemonic role of the prefrontal cortex, including attentional control of sensory processing, integration of information from different domains, stimulus selection and monitoring of information held in memory. Our own data argue against a modulatory view of the prefrontal cortex and suggest that processes supporting working memory are distributed along ventral and dorsal lateral prefrontal cortex. PMID- 16352403 TI - Plasmacytoid monocytes in cat scratch disease with special reference to the histological diversity of suppurative lesions. AB - It has been suggested that plasmacytoid monocytes (PMOs) play an essential role in T-cell-dependent immune response. Indeed, numerous PMOs are found in close topographical association with epithelioid cell granulomas in hypersensitivity type granulomas, such as tuberculosis and sarcoidosis. The key pathologic process in cat scratch disease (CSD) usually involves a B-cell-associated granulomatous reaction. Histologically, CSD appears to exhibit a histopathologic diversity, including suppurative lesions without epithelioid cell granulomas (early lesion), in which the microabscesses were surrounded by monocytoid B-cells (MBCs), suppurative granulomas containing MBCs (intermediate lesion), and suppurative granulomas without MBCs (late lesion). However, the presence or absence of PMO in CSD has not been studied previously. We examined 14 cases of CSD. In early lesions, numerous clusters of PMO were detected in the MBCs. In intermediate lesions, both MBCs and PMOs were found to be decreased in number, while late lesions contained no or only a few MBCs and PMOs. Overall, these findings suggest that PMOs may play a role in MBC-associated granulomatous response and in hypersensitivity granulomatous response. Moreover, the association with MBCs and PMOs indicates a functional relationship of MBCs with PMOs in the formation of suppurative lesions in CSD. PMID- 16352404 TI - Functional analysis of bovine herpesvirus 1 (BHV-1) genes expressed during latency. AB - Bovine herpes virus 1 (BHV-1) establishes latency in sensory neurons of trigeminal ganglia (TG), and germinal centers of pharyngeal tonsil. Periodically BHV-1 reactivates from latency, virus is shed, and consequently virus transmission occurs. Two transcripts, the latency related (LR) RNA and ORF-E, are abundantly expressed in TG of latently infected cattle. A LR mutant strain of BHV 1 was constructed that contains stop codons near the beginning of the LR-RNA. The LR mutant virus does not express two proteins encoded by the LR gene, or reactivate from latency suggesting that LR protein expression regulates the latency-reactivation cycle. Higher levels of apoptosis occur in TG of calves infected with the LR mutant versus wild type BHV-1 indicating that the anti apoptotic properties of the LR gene regulate the latency-reactivation cycle. The LR gene also inhibits bICP0 expression and mammalian cell growth, but these functions do not require LR protein expression. In contrast, the ability of the LR gene to inhibit apoptosis appears to require LR protein expression. A small open reading frame (ORF-E) that is located within the LR promoter is expressed in the nucleus of neuroblastoma cells. We predict that the LR gene and ORF-E regulate the BHV-1 latency-reactivation cycle. PMID- 16352405 TI - Host cell targets of immediate-early protein BICP22 of bovine herpesvirus 1. AB - The immediate-early (IE) protein BICP22 of bovine herpesvirus 1 (BHV-1) acts as transrepressor protein on viral promoters of different kinetic classes. In the present work, we looked for host cell targets of BICP22 using a yeast two-hybrid system and identified seven candidates: (1) JIK, a serine/threonine kinase of the sterile 20 protein (STE20) family that inhibits stress-related pathways; (2) cAMP response element binding protein-like 2 (CREBL2), which in its bZip domain shares homology with CREB, modulating transcription of cAMP responsive genes; (3) DNA dependent ATPase and helicase (ATRX), a protein of the SNF2 family altering nucleosome structure; (4) scaffold attachment factor B (SAF-B), which helps to organize chromatin into topologically separated loops; (5) peptidylglycine alpha amidating monooxygenase COOH-terminal interactor protein 1 (PAMCIP1), involved in regulation of the secretory pathway in the perinuclear area; (6) zinc finger protein (ZNF38) found in proliferating cells and possibly associated with meiosis in male and female gametogenesis; (7) FLJ22709, hypothetical protein conserved among various species, containing an occludin/ELL domain. To confirm some of the interactions by confocal fluorescence microscopy, BICP22 was tagged with red fluorescent protein in an amplicon, and selected target sequences were tagged with green fluorescent protein in plasmid expression vectors. Upon amplicon transduction of Vero cells and plasmid transfection, CREBL2 and ZNF38 both colocalized with BICP22 in distinct nuclear domains. PMID- 16352406 TI - Formation of phenytoin nanoparticles using rapid expansion of supercritical solution with solid cosolvent (RESS-SC) process. AB - Nanoparticles are of significant importance in drug delivery. Rapid expansion of supercritical solution (RESS) process can produce pure and high-quality drug particles. However, due to extremely low solubility of polar drugs in supercritical CO(2) (sc CO(2)), RESS has limited commercial applicability. To overcome this major limitation, a modified process rapid expansion of supercritical solution with solid cosolvent (RESS-SC) is proposed which uses a solid cosolvent. Here, the new process is tested for phenytoin drug using menthol solid cosolvent. Phenytoin solubility in pure sc CO(2) is only 3 micromol/mol but when menthol solid cosolvent is used the solubility is enhanced to 1,302 micromol/mol, at 196 bar and 45 degrees C. This 400-fold increase in the solubility can be attributed to the interaction between phenytoin and menthol. Particle agglomeration in expansion zone is another major issue with conventional RESS process. In proposed RESS-SC process solid cosolvent hinders the particle growth resulting in the formation of small nanoparticles. For example, the average particle size of phenytoin in conventional RESS process is 200 nm whereas, with RESS-SC process, the average particle size is 120 nm, at 96 bar and 45 degrees C. Similarly at 196 bar and 45 degrees C, 105 nm average particles were obtained by RESS and 75 nm average particles were obtained in RESS-SC process. The particles obtained were characterized by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS) and differential scanning calorimetery (DSC) analyses. Phenytoin nanoparticle production rate in RESS-SC is about 400-fold more in comparison to that in RESS process. PMID- 16352407 TI - Evaluation of an integrated in vitro-in silico PBPK (physiologically based pharmacokinetic) model to provide estimates of human bioavailability. AB - PK express module is a physiologically based model of first pass metabolism, which integrates in vitro data with an in silico physiologically based pharmacokinetic (PBPK) model to predict human bioavailability (F(H)). There are three required inputs: FDp (Fraction dose absorbed, final parameter from iDEA absorption module), protein binding (fu) and disappearance kinetics in human hepatocytes. Caco-2 permeability, aqueous solubility (at multiple pH's), estimated dose and chemical structure are inputs required for the estimation of FDp (Norris et al., 2000; Stoner et al., 2004) and were determined for all compounds in our laboratory or obtained from literature. Protein binding data was collected from literature references and/or Pfizer database. Human hepatocyte data was generated in-house using an automated human hepatocyte method (using Tecan Genesis Workstation) as described previously (). Sixteen compounds (commercial and Pfizer compounds) were chosen to evaluate the PK express model and the bioavailability predicted from the module was compared with known clinical endpoints. For majority of the 16 compounds (approximately 80%), the PK express model F(H) values were comparable to the known human bioavailability (F(H)) (within 23.7 units of the known human (true) F, except for PF 3, PF 4, PF 6). In conclusion, the PK express model integrates a number of key readily available discovery parameters and provides estimates of human performance by integrating in silico and experimental variables built on a physiological based pharmacokinetic model. Information from this model in conjunction with other ADME data (e.g., P450 inhibition) will enable progression of most promising compounds for further in vivo PK and/or efficacy studies. PMID- 16352408 TI - What shall we do with the drunken sailor? Effects of alcohol on the performance of ship operators. AB - The purpose of this study was to specify the effects of alcohol on the performance of ship operators as a contribution to the development of new strategies against the risks of alcohol in water traffic. The nautical performance of 21 captains before and after alcohol consumption was assessed on a ship piloting simulator. The simulated scenarios represented passages of a container vessel through the German Bight. Performance was examined by nautical instructors according to standardised protocols. Mean (S.D.) blood alcohol concentrations (BACs) of 0.100 (0.024) g/dl before and 0.100 (0.017) g/dl after the performance trial resulted in striking effects on the nautical performance. The categories most severely affected were foresight and analysis of situation (impairment in 18 of 21 cases), concentration (impairment in 16 of 21 cases), accurateness, risk disposition and navigation (impairment in 15 of 21 cases). Chart work, preparation and communication were impaired in 12, 11 and 10 of 21 cases, respectively. None of the participants were capable to operate the simulated ship with an adequate safety after ingestion of alcohol. From these findings, and in consideration of the well-established impairment of a multitude of mental and physical functions by alcohol, it can be concluded that even low BACs bear high risks in water traffic, a concentration above 0.1 g/dl will hinder a sufficiently safe performance of ship operators. This should be considered in alcohol education and legislation. PMID- 16352410 TI - Recent female mouse models displaying advanced reproductive aging. AB - Reproductive senescence occurs in all female mammals with resultant changes in numerous body functional systems and several important features may be species specific. Those features that appear to parallel human menopause and aging include general similarity of hormone profiles across the menopausal transition, progression to cycle termination through irregular cycles, declining fertility with age, disturbances in thermogenesis, age-related gains in body weight, fat distribution and disposition towards metabolic syndrome. Structural and hormonal changes in the brain and ovary play a critical role in determining the onset of reproductive senescence. The short life span of rodents such as mice (compared to humans) and the ability to generate specific and timed gene deletions, provide powerful experimental paradigms to understand the molecular and functional changes that precede and follow the loss of reproductive capacity. In theory, any manipulation that compromises ovarian function either partly or totally would impact reproductive events at various levels followed by other dysfunctions. In this article, we provide an overview of three mouse models for the study of female reproductive aging. They are derived from different strategies and their age related phenotypes have been characterized to varying degrees. The follitropin receptor knockout (FORKO) mouse, in its null and haploinsufficient state as well as the dioxin/aryl hydrocarbon receptor (AhR) knockout mouse, serve as two examples of single gene deletions. A third model, using administration of a chemical toxicant such as 4-vinylcyclohexene diepoxide (VCD) in the adult state, produces ovarian deficiencies accompanied by aging changes. These will serve as useful alternatives to previously used radical ovariectomy in young adults. It is anticipated that these new models and more that will be forthcoming will extend opportunities to understand reproductive aging and resolve controversies that abound on issues related to benefits and risks of hormone replacement therapy or other modalities for improving quality of life. PMID- 16352411 TI - Valsalva and gravitational variability of the internal jugular vein and common femoral vein: ultrasound assessment. AB - PURPOSE: Central venous cannulation via the common femoral vein is an important starting point for many interventions. The purpose of this study was to determine the optimum conditions for cannulation of the femoral vein and to compare these with the relative changes in the internal jugular vein. METHODS: High-resolution 2D ultrasound was utilised to determine variability of the calibre of the femoral and internal jugular veins in 10 healthy subjects. Venous diameter was assessed during the Valsalva manoeuvre and in different degrees of the Trendelenburg position. RESULTS: The Valsalva manoeuvre significantly increased the size of the femoral and internal jugular veins. There was a relatively greater increase in femoral vein diameter when compared with the internal jugular vein of 40 and 29%, respectively. Changes in body inclination (Trendelenburg position) did not significantly alter the luminal diameter of the femoral vein. However, it significantly increased internal jugular vein diameter. CONCLUSIONS: Femoral vein cannulation is augmented by the Valsalva manoeuvre but not significantly altered by the gravitational position of the subject. PMID- 16352412 TI - Comparative sequence analysis of the internal transcribed spacer 1 of Ochrobactrum species. AB - The internal 16S/23S rDNA (rrs/rrl) internal spacer region 1 (ITS1) of 54 Ochrobactrum strains and close relatives was analysed. Separation of ITS1 containing PCR products by gel-electrophoresis, DGGE, cloning and sequencing revealed ITS1 length and sequence heterogeneity. We found up to 5 different allelic ITS1 stretches within a single strain (Ochrobactrum intermedium LMG 3301T), and 2-3 different ITS1 alleles in O. tritici. Within ITS1, ITS1c, being part of the conserved double-stranded rrn processing stem dsPS1, produced the most reliable segment tree. The overall ITS1, ITS1c and rrs phylogenetic tree topologies were generally consistent, but there was evidence for horizontal rrn (segment) transfer in O. tritici LMG 2134 (formerly O. anthropi). Good correlations were found between ITS1, ITS1c and rrs sequence similarity and DNA DNA hybridization values indicating that phylogenetic analysis of ITS1 and ITS1c both can be used to preliminarily deduce the phylogenetic affiliation if HGT was excluded. Strains sharing > 96.19% ITS1c (> 95.11% ITS1) similarity fell within a species, and < or = 68.42% ITS1c (< or = 70.33% ITS1) similarity outside a genus. Both ITS1 and ITS1c analysis resolved microdiversity more profoundly than rrs analysis and revealed clades (genomovars) within O. anthropi that were also produced in rep cluster analysis. There was no evidence for habitat-specific ITS1 genomovars within Ochrobactrum species. Diversity of Ochrobactrum was higher in soil than at the rhizoplane below and at the species level. Isolates from soil contained only 1 rrn type whereas isolates from human clinical, animal and rhizoplane specimens could contain more. PMID- 16352413 TI - Evaluation of chemometric techniques to select orthogonal chromatographic systems. AB - Several chemometric techniques were compared for their performance to determine the orthogonality and similarity between chromatographic systems. Pearson's correlation coefficient (r) based color maps earlier were used to indicate selectivity differences between systems. These maps, in which the systems were ranked according to decreasing or increasing dissimilarities observed in the weighted-average-linkage dendrogram, were now applied as reference method. A number of chemometric techniques were evaluated as potential alternative (visualization) methods for the same purpose. They include hierarchical clustering techniques (single, complete, unweighted-average-linkage, centroid and Ward's method), the Kennard and Stone algorithm, auto-associative multivariate regression trees (AAMRT), and the generalized pairwise correlation method (GPCM) with McNemar's statistical test. After all, the reference method remained our preferred technique to select orthogonal and identify similar systems. PMID- 16352414 TI - Development of immunoaffinity solid phase microextraction probes for analysis of sub ng/mL concentrations of 7-aminoflunitrazepam in urine. AB - We report on the development of solid phase microextraction probes for drug analysis, prepared with antibodies specific for benzodiazepines covalently immobilized to the surface. In the technique, immobilized antibody probes are exposed to a sample containing the drug for 30 min. Extracted drugs are subsequently desorbed from the probes in 500 microL of methanolic desorption solution, which is dried, reconstituted in a small volume of injection solution and analysed by LC-MS/MS. The antibodies were characterized both before and after immobilization, to facilitate the rational selection of antibodies for such analyses. Polyclonal and monoclonal antibodies were compared as was the impact of affinity purification of the polyclonal antibody to isolate the drug-specific fraction. The probes were evaluated for utility in analyzing 7-aminoflunitrazepam at sub ng/mL concentrations in urine, which is expected to be found several days after a single oral dose of 2 mg of flunitrazepam. Such analyses are required in monitoring for abuse of this drug, both in terms of 'club drug' use and in cases of drug-facilitated sexual assault. In these cases drug concentrations in blood and urine are much lower than in chronic abuse cases and are difficult to analyse by conventional methods. The method developed has a limit of detection of 0.02 ng/mL, with accuracy ranging from 1% to 27% and precision (% R.S.D.) ranging from 2% to 10% between the lower and upper limits of quantitation for the analysis of 7-aminoflunitrazepam in urine. The dynamic range of the method is from 0.02 ng/mL, which is limited by the instrument sensitivity, to 0.5 ng/mL, which is approaching the capacity of the probes. This would allow for quantitative analysis of samples at concentrations below that measurable by many other methods for general benzodiazepines analysis from urine, and a highly selective screen for samples at higher concentrations. The method has similar limits of detection to the most sensitive literature methods specifically designed for such analysis but with the advantage of significantly simplified sample preparation. This simplification makes the technique more amenable for use by both professionals and non-professionals. PMID- 16352415 TI - Comparison of UV spectrophotometric method and high performance liquid chromatography for the analysis of flunarizine and its application for the dissolution test. AB - This study aimed to develop a simple UV spectrophotometric method for the analysis and the dissolution test of flunarizine in capsules. The UV absorbance was both measured directly and by the first derivative measurements at 254 and 268 nm, respectively. The developed methods were validated for their linearity, accuracy, precision, limit of detection (LOD) and limit of quantitation (LOQ) in comparison with the reported HPLC method. The UV spectrophotometric method illustrated excellent linearity (r2 > 0.9999) in the concentration range of 6-24 microg/mL. Precision (%R.S.D. < 1.50) and recoveries were good (%R > 99.62). The LOD of direct UV and first derivative measurements were 0.09 and 0.84 microg/mL, respectively, and the LOQ were 0.26 and 2.55 microg/mL, respectively. Results from the assay of flunarizine in capsules by the UV spectrophotometric methods, both direct and first derivative measurements were not significantly different from those of the HPLC method (P > 0.05). Additionally, the method was successfully used for the dissolution test of flunarizine capsule and was found to be reliable, simple, fast, and inexpensive. PMID- 16352416 TI - A hybrid generative and predictive model of the motor cortex. AB - We describe a hybrid generative and predictive model of the motor cortex. The generative model is related to the hierarchically directed cortico-cortical (or thalamo-cortical) connections and unsupervised training leads to a topographic and sparse hidden representation of its sensory and motor input. The predictive model is related to lateral intra-area and inter-area cortical connections, functions as a hetero-associator attractor network and is trained to predict the future state of the network. Applying partial input, the generative model can map sensory input to motor actions and can thereby perform learnt action sequences of the agent within the environment. The predictive model can additionally predict a longer perception- and action sequence (mental simulation). The models' performance is demonstrated on a visually guided robot docking manoeuvre. We propose that the motor cortex might take over functions previously learnt by reinforcement in the basal ganglia and relate this to mirror neurons and imitation. PMID- 16352417 TI - Comparison of recent methods for inference of variable influence in neural networks. AB - Neural networks (NNs) belong to 'black box' models and therefore 'suffer' from interpretation difficulties. Four recent methods inferring variable influence in NNs are compared in this paper. The methods assist the interpretation task during different phases of the modeling procedure. They belong to information theory (ITSS), the Bayesian framework (ARD), the analysis of the network's weights (GIM), and the sequential omission of the variables (SZW). The comparison is based upon artificial and real data sets of differing size, complexity and noise level. The influence of the neural network's size has also been considered. The results provide useful information about the agreement between the methods under different conditions. Generally, SZW and GIM differ from ARD regarding the variable influence, although applied to NNs with similar modeling accuracy, even when larger data sets sizes are used. ITSS produces similar results to SZW and GIM, although suffering more from the 'curse of dimensionality'. PMID- 16352418 TI - Lee-Associator-a chaotic auto-associative network for progressive memory recalling. AB - In the past few decades, neural networks have been extensively adopted in various applications ranging from simple synaptic memory coding to sophisticated pattern recognition problems such as scene analysis and robot vision. Moreover, current studies on neuroscience and physiology have reported that in a typical scene segmentation problem our major senses of perception (e.g. vision, olfaction, etc.) are highly chaotic and involved non-linear neural dynamics and oscillations. In this paper, the author proposes an innovative chaotic neural oscillator-namely the Lee-oscillator (Lee's Chaotic Neural Oscillator) to provide a chaotic neural coding and information processing scheme. To illustrate the capability of Lee-oscillators upon pattern association, a chaotic auto associative network, namely Lee-Associator (Lee's Chaotic Auto-associator) is constructed. Different from classical auto-associators such as the celebrated Hopfield network, which provides time-independent and static pattern association scheme, the Lee-Associator provides a remarkable progressive memory association scheme (what the author called 'Progressive Memory Recalling Scheme, PMRS') during the chaotic memory association. This is exactly consistent with the latest research in psychiatry and perception psychology on dynamic memory recalling schemes, as well as the implications and analogues to human perception as illustrated by the remarkable Rubin-vase experiment on visual psychology. PMID- 16352419 TI - Energy efficient and robust rhythmic limb movement by central pattern generators. AB - Humans show great energy efficiency and robustness in rhythmic tasks, such as walking and arm swinging. In this study a mathematical model of rhythmic limb movement is presented, which shows that tight local coupling of Central Pattern Generators (CPGs) to limbs could explain part of this behavior. Afferent feedback to flexor and extensor centers of the CPG is crucial in providing energy efficiency by means of resonance tuning. Feedback of positional information provides resonance tuning above the endogenous frequency of the CPG. Integral feedback provides resonance tuning at and below the endogenous frequency. Feedback of velocity information is necessary to compensate for the time delay in the loop, coupling limb to CPG; without velocity feedback bi-stability occurs and resonance tuning is not possible at high movement frequencies. The concepts of energy efficient and robust control of rhythmic limb movements are also applicable to robotics. It is the first CPG model, which provides resonance tuning at natural limb frequencies above and below its endogenous frequency. PMID- 16352420 TI - Non-surgical treatment modalities of facial photodamage: practical knowledge for the oral and maxillofacial professional. AB - With the increasing interest in cosmetic procedures, oral and maxillofacial surgeons are being asked not only to improve oral health and aesthetics but to extend their expertise to provide advice on improving the overall appearance of the face. For the discerning patient, improving overall facial skin appearance is becoming an integral part of the process of surgical cosmetic procedures. Here, some of the non-surgical options available for the treatment of photodamaged skin are reviewed and an overview of the specific treatments in this category provided. Sun avoidance and protection from harmful rays with appropriate sunscreens are primary to maintaining healthy skin and appearance. Among treatment options, topical treatments with preparations such as retinoids, alpha hydroxy acids and antioxidants have been shown to provide some benefit and are relatively easy to use albeit with appropriate precautions and professional guidance. As a second-level option, facial rejuvenation procedures such as botulinum toxin injection, soft tissue augmentation with collagen or hyaluronic acid gel, skin resurfacing, use of chemical peels, dermabrasion and laser resurfacing procedures can be used but require administration by qualified practitioners. Overall, these treatments may be used to complement rehabilitative, reconstructive, or cosmetic oral and maxillofacial surgery to further improve and complement surgical results. PMID- 16352421 TI - Differential role of MAPK signaling in human dendritic cell maturation and Th1/Th2 engagement. AB - Dendritic cells (DCs) are potent antigen-presenting cells that can stimulate resting T cells in the primary immune response. During the maturation process, immature DCs lose their ability to internalize antigens and they acquire the capacity to present antigens to naive T cells. Many observations have suggested that distinct DC subsets might differentially regulate Th responses. However, recent reports suggest that specific subsets of either murine or human DCs cultured in vitro with different stimuli respond with great plasticity in terms of both gene expression and cytokine secretion. Thus, the microenvironment of DCs may determine the nature of mature DCs and the subsequent immune response. The mechanism by which the character of DCs is determined is unknown. The in vitro maturation process of human monocyte-derived DCs (MoDC) can be initiated by various stimuli. Many stimuli induce phosphorylation of extracellular signal regulated kinases (ERK), c-Jun N-terminal kinases (JNK), and p38 MAPK in DCs during maturation. Such kinase-specific inhibitors help to reveal the functions of MAPKs in the maturation of human MoDCs. Recent studies suggest that three MAPK signaling pathways differentially regulate all aspects of phenotypic maturation, cytokine production, and functional maturation of MoDCs. Thus, distinct maturation of DCs may be induced by modulating the balance of phosphorylation of the three MAPKs. In this review, we summarize the role of MAPK signaling pathways in the maturation of human MoDCs. PMID- 16352422 TI - Transferable DHA-1 cephalosporinase in Escherichia coli. AB - Three Escherichia coli isolates resistant to third-generation cephalosporins but negative for extended-spectrum beta-lactamase production were isolated from hospitalised patients in Zagreb, Croatia, during June 2003 to February 2004. Resistance was due to the inducible production of a DHA-1 cephalosporinase. Each isolate contained an integron-associated bla(DHA-1)-ampR sequence carried by similar-sized plasmids, of which one was self-transferable. Serotyping and polymerase chain reaction typing using ERIC2 primer indicated that the isolates were distinct. This is the first description of DHA beta-lactamase production in E. coli. PMID- 16352423 TI - Targeting antioxidants to mitochondria: a new therapeutic direction. AB - Mitochondria play an important role in controlling the life and death of a cell. Consequently, mitochondrial dysfunction leads to a range of human diseases such as ischemia-reperfusion injury, sepsis, and diabetes. Although the molecular mechanisms responsible for mitochondria-mediated disease processes are not fully elucidated yet, the oxidative stress appears to be critical. Accordingly, strategies are being developed for the targeted delivery of antioxidants to mitochondria. In this review, we shall briefly discuss cellular reactive oxygen species metabolism and its role in pathophysiology; the currently existing antioxidants and possible reasons why they are not effective in ameliorating oxidative stress-mediated diseases; and recent developments in mitochondrially targeted antioxidants and their future promise for disease treatment. PMID- 16352424 TI - Efficiency of 1H --> 31P NMR cross-polarization in bone apatite and its mineral standards. AB - Human bone mineral was studied using solid-state 31P NMR with cross-polarization (CP) from protons. The CP efficiency was determined for trabecular and cortical bone tissue from human adults and compared with synthetic mineral standards. The study shows the similarity between carbonatoapatite of type B and bone mineral as shown by their CP behaviour. The method can be used for the characterization of synthetic apatite-based implant materials. PMID- 16352425 TI - Bioactive heparin immobilized onto microfluidic channels in poly(dimethylsiloxane) results in hydrophilic surface properties. AB - A new composition of heparin coating for microfluidic systems made out of poly(dimethylsiloxane) (PDMS) was developed and evaluated. The coating that consists of a conditioning polyamine layer followed by two heparin/glutaraldehyde layers, resulted in channel surfaces with sufficient wettability to obtain flow of human normal plasma by capillary force alone. Hydrophilic channel walls are a desirable characteristic in microfluidic devices, since alternative pumping mechanisms must otherwise be included into the system. The immobilized heparin showed high antithrombin-binding capacity and a low degree of blood-material interaction. Plasma in contact with heparin-coated PDMS formed no detectable fibrin in a spectrophotometric assay by which plasma in contact with non-treated PDMS showed complete coagulation. The quartz crystal microbalance technique with energy dissipation monitoring (QCM-D) was utilized to obtain detailed information regarding adsorption kinetics and structural properties of the different layers composing the heparin coating. PMID- 16352426 TI - The power of the pump: mechanisms of action of P-glycoprotein (ABCB1). AB - Members of the superfamily of ATP-binding cassette (ABC) transporters mediate the movement of a variety of substrates including simple ions, complex lipids and xenobiotics. At least 18 ABC transport proteins are associated with disease conditions. P-glycoprotein (Pgp, ABCB1) is the archetypical mammalian ABC transport protein and its mechanism of action has received considerable attention. There is strong biochemical evidence that Pgp moves molecular cargo against a concentration gradient using the energy of ATP hydrolysis. However, the molecular details of how the energy of ATP hydrolysis is coupled to transport remain in dispute and it has not been possible to reconcile the data from various laboratories into a single model. The functional unit of Pgp consists of two nucleotide binding domains (NBDs) and two trans-membrane domains which are involved in the transport of drug substrates. Considerable progress has been made in recent years in characterizing these functionally and spatially distinct domains of Pgp. In addition, our understanding of the domains has been augmented by the resolution of structures of several non-mammalian ABC proteins. This review considers: (i) the role of specific conserved amino acids in ATP hydrolysis mediated by Pgp; (ii) emerging insights into the dimensions of the drug binding pocket and the interactions between Pgp and the transport substrates and (iii) our current understanding of the mechanisms of coupling between energy derived from ATP binding and/or hydrolysis and efflux of drug substrates. PMID- 16352427 TI - An intelligent tutoring system that generates a natural language dialogue using dynamic multi-level planning. AB - OBJECTIVE: The objective of this research was to build an intelligent tutoring system capable of carrying on a natural language dialogue with a student who is solving a problem in physiology. Previous experiments have shown that students need practice in qualitative causal reasoning to internalize new knowledge and to apply it effectively and that they learn by putting their ideas into words. METHODS: Analysis of a corpus of 75 hour-long tutoring sessions carried on in keyboard-to-keyboard style by two professors of physiology at Rush Medical College tutoring first-year medical students provided the rules used in tutoring strategies and tactics, parsing, and text generation. The system presents the student with a perturbation to the blood pressure, asks for qualitative predictions of the changes produced in seven important cardiovascular variables, and then launches a dialogue to correct any errors and to probe for possible misconceptions. The natural language understanding component uses a cascade of finite-state machines. The generation is based on lexical functional grammar. RESULTS: Results of experiments with pretests and posttests have shown that using the system for an hour produces significant learning gains and also that even this brief use improves the student's ability to solve problems more then reading textual material on the topic. Student surveys tell us that students like the system and feel that they learn from it. The system is now in regular use in the first-year physiology course at Rush Medical College. CONCLUSION: We conclude that the CIRCSIM-Tutor system demonstrates that intelligent tutoring systems can implement effective natural language dialogue with current language technology. PMID- 16352428 TI - Effect of fish oil on offensive and defensive factors in gastric ulceration in rats. AB - The effect of fish oil (FO) derived from Scomberoides commersonianus containing omega-3 polyunsaturated fatty acids was studied on gastric ulcers and as well as on offensive and defensive factors in gastric mucosal damage, following experimental gastric ulceration. FO significantly reduced the severity of ulceration in gastric ulcers induced by aspirin, cold-restraint stress (CRS), alcohol, and pylorus ligation. The results also indicated the potentiality of FO in maintaining the integrity of gastric mucosa by virtue of its effect on both offensive and defensive gastric mucosal factors. It decreased the offensive acid pepsin secretion and augmented the defensive factors like mucin secretion, cellular mucus and life span of mucosal cells following pylorus ligation. FO significantly increased activity of anti-oxidant enzymes (catalase and glutathione peroxidase) and decreased lipid peroxidation in gastric mucosa of CRS rats. The study indicates the beneficial role of FO in gastric ulceration by inhibition of offensive mucosal factors and oxidative stress, and augmentation of defensive mucosal factors. PMID- 16352429 TI - Organizing moving groups during morphogenesis. AB - The directed migration of cells drives the formation of many complex organ systems. Although in this morphogenetic context cells display a strong preference for migrating in organized, cohesive groups, little is known about the mechanisms that coordinate their movements. Recent studies on several model systems have begun to dissect the organization of these migrating tissues in vivo and have shown that cell guidance is mediated by a combination of chemical and mechanical cues. PMID- 16352430 TI - Biosensing approach for alcohol determination using immobilized alcohol oxidase. AB - Alcohol oxidase (AOD) was immobilized in polypyrrole (PPy) and a random copolymer containing 3-methylthienyl methacrylate and p-vinylbenzyloxy poly(ethyleneoxide) matrices. Immobilization of enzyme was performed via entrapment in conducting polymers during electrochemical polymerization of pyrrole through the thiophene moiety of the copolymer. Three different alcohols, namely methanol, ethanol and n propanol, were used as substrates. Maximum reaction rates, Michaelis-Menten constants, optimum temperature and pH values, operational stabilities and shelf life of the enzyme electrodes were investigated. PMID- 16352432 TI - Preparation and preliminary biological evaluation of a (166)Ho labeled polyazamacrocycle for possible use as an intravascular brachytherapy (IVBT) agent. AB - (166)Ho can be considered as a potential radionuclide for intravascular brachytherapy (IVBT) using liquid-filled balloons owing to its suitable nuclear decay characteristics. The possibility of producing (166)Ho with adequate specific activity using moderate flux reactors and natural holmium target makes it an attractive alternative of (188)Re for developing IVBT agents. Keeping in mind the high thermodynamic stability of lanthanide complexes with polyazamacrocycles, (166)Ho complex of 1,4,8,11-tetraazacyclotetradecane-1,4,8,11 tetraacetic acid (TETA) was prepared and studied for its suitability as a possible agent for IVBT. (166)Ho was produced with adequate specific activity and high radionuclidic purity by irradiating natural Ho(2)O(3) powder. TETA was synthesized by a single step procedure using cyclam as the starting material. (166)Ho-TETA complex was prepared with excellent radiochemical purity and the complex was found to retain its stability for 7 days at room temperature. Biodistribution studies carried out in Wistar rats showed major renal clearance of the injected activity with almost no retention in any of the vital organ/tissue. PMID- 16352431 TI - Reduced bone tunnel enlargement post hamstring ACL reconstruction with poly-L lactic acid/hydroxyapatite bioabsorbable screws. AB - Bone tunnel enlargement following anterior cruciate ligament (ACL) reconstruction can complicate revision surgery. This study compared postoperative tibial tunnel widening in patients who underwent arthroscopically assisted, single-incision, four-strand hamstring ACL reconstruction using a poly-L-lactic acid/hydroxyapatite blend (PLLA+HA) bioabsorbable interference screw for tibial fixation, with those in whom a plain poly-L-lactic acid (PLLA) screw was used. Thirty-four patients (13 with PLLA+HA tibial interference screw fixation and 21 with plain PLLA tibial interference screws) underwent a spiral CT scan to assess maximum tibial tunnel cross-sectional area at an average of 28.7 months follow up. An assessment of tunnel wall sclerosis adjacent to the screw (cortication) was also made. The two groups were well matched for age, sex and graft diameters. Mean tibial tunnel enlargement in patients with PLLA+HA screws was 29.9% at average 30.9 months follow-up compared with 46% in patients with plain PLLA screw at an average 26.5 months follow-up (p=0.03). The tunnel wall adjacent to the screw appeared corticated in only 21% of patients with PLLA+HA screws (p=0.02) compared with 73% of patients with PLLA screws. The blending of HA with PLLA appears to reduce postoperative tunnel widening, and the reduced tunnel wall sclerosis seen postoperatively may indicate improved screw incorporation. PMID- 16352433 TI - Immunoreactivity assay for alpha-particle emitting monoclonal antibody constructs. AB - Clinical trials using alpha-particle radiolabeled antibodies require a rapid and reproducible assay of the immunoreactivity of drugs. While live cell assays are typically used to determine the immunoreactive drug fraction, a fixed cell assay may replace the traditional live cell assay and offer the advantages of rapidity, easy availability and consistency for qualifying drugs for preclinical or clinical studies. We have identified optimal cell fixation and immunoreactivity assay conditions and have validated them by performing the fixed-cell assay in clinical trials. PMID- 16352434 TI - Intraventricular administration of hepatocyte growth factor treats mouse communicating hydrocephalus induced by transforming growth factor beta1. AB - Communicating hydrocephalus may occur spontaneously in elderly patients or occur as a complication of meningitis or intracranial hemorrhage, typically as a result of fibrosis along the route of cerebrospinal fluid (CSF) flow. Hepatocyte growth factor (HGF) has anti-fibrotic properties and is a promising candidate for the treatment of various fibrotic diseases. Thus, the goal of this study was to examine the effect of exogenous HGF (30 microg of human recombinant (hr) HGF intraventricularly for 7 or 14 days) in a model of hr transforming growth factor beta1-induced communicating hydrocephalus in C57BL/6 mice. HGF treatment resulted in a reduction of ventriculomegaly, as demonstrated by magnetic resonance imaging, and improved spatial memory. Further, ink passage test demonstrated improvement of normalized CSF in flow in mice receiving HGF treatment as opposed to delayed CSF flow in the hydrocephalic mice at baseline. Finally, histological examination in hydrocephalic mice undergoing HGF treatment revealed reduction of collagen fibers in the meninges and normalization of their structures. These results indicate that exogenous HGF may be of utility in the treatment of hydrocephalus in humans. PMID- 16352435 TI - Treatment of vascular lesions in the head and neck using Nd:YAG laser. AB - INTRODUCTION: Vascular lesions in the head and neck region, including both haemangiomas and vascular malformations, are common and many different treatment modalities have been used for their removal. In the past decade, the Nd:YAG laser has emerged as a new mode of treatment for vascular lesions, and the purpose of this paper was to determine its clinical value. PATIENTS AND METHODS: A prospective study was conducted in 111 patients with vascular lesions in the head and neck region. They were treated with the Nd:YAG laser by photocoagulation. Of these, 96 had small lesions, with surface diameters of less than 3 x 3cm(2), and 5 had large lesions, with surface diameters of more than 3 x 3cm(2). The patients were all followed-up carefully until complete healing was recorded, along with any complications. RESULTS: In both groups of patients, tissue sloughing occurred within 2-3 days. Healing time in small lesions was 2-3 weeks, and in large lesions 3-4 weeks. Three patients with small lesions and one patient with a large lesion experienced minor complications. CONCLUSION: The Nd:YAG laser is a safe and effective tool for treating vascular lesions. PMID- 16352436 TI - High amplitude short time excitation: a method to form and detect low mass product ions in a quadrupole ion trap mass spectrometer. AB - Collision induced dissociation (CID) in a quadrupole ion trap mass spectrometer using the conventional 30 ms activation time is compared with high amplitude short time excitation (HASTE) CID using 2 ms and 1 ms activation times. As a result of the shorter activation times, dissociation of the parent ions using the HASTE CID technique requires resonance excitation voltages greater than conventional CID. After activation, the rf trapping voltage is lowered to allow product ions below the low mass cut-off to be trapped. The HASTE CID spectra are notably different from those obtained using conventional CID and can include product ions below the low mass cut-off for the parent ions of interest. The MS/MS efficiencies of HASTE CID are not significantly different when compared with the conventional 30 ms CID. Similar results were obtained with a two dimensional (linear) ion trap and a three-dimensional ion trap. PMID- 16352437 TI - A parallel approach to post source decay MALDI-TOF analysis. AB - We present a novel enhancement to matrix-assisted laser desorption ionization (MALDI) post-source decay (PSD) analysis whereby fragment ions from multiple precursor ions are acquired into the same spectrum without employing a timed ion gate to preselect each parent ion. Fragment ions are matched to their corresponding precursor ions by comparing spectra acquired at slightly different reflectron electric fields. By measuring the difference in time-of-flight (TOF) between the two spectra for each fragment, it is possible to calculate the mass of the fragment ion and its parent. This new "parallel PSD" technique reduces analysis time and consumes less sample than conventional PSD, which requires an ion gate for serial preselection of precursor ions. PMID- 16352438 TI - Gas phase H/D exchange of sodiated amino acids: why do we see zwitterions? AB - The gas-phase interaction of sodiated amino acids and sodiated amino acid methyl esters with various deuterium donors is investigated by combining results of H/D exchange reactions with those from density functional theory and molecular dynamics calculations. Discrepancy between experimentally and theoretically obtained structures for sodium cationized amino acids is explained by deuterium donor caused perturbation of the most stable amino acid conformation. Detailed study of H/D exchange mechanism on sodiated amino acids shows that the H/D exchange reaction is preceded by a multistep quasi-isoenergetic transition (perturbation) from a charge solvated to zwitterionic structure in the amino acid. Although the computation refers to the system AlaNa(+) and D(2)O, these mechanisms apply to all amino acids, except those where a functional side-chain group takes part in the perturbation process. The suggested perturbation mechanism applies also for other deuterium donors such as CD(3)OD or even ND(3) and indicates that a single water molecule suffices to convert the sodiated amino acid from charge solvated to zwitterionic form. PMID- 16352440 TI - In situ generation of HCN for mass spectrometric studies. AB - Hydrogen cyanide (HCN) for use in ion preparation can be generated in the gas phase by the neutral-neutral reaction of trimethylsilyl cyanide (Me(3)SiCN) and water in a flowing afterglow mass spectrometer. We demonstrate that the approach can be used to generate a wide range of HCN solvated ions such as F(-)(HCN), Cl( )(HCN), CN(-)(HCN), PhNO(2)(.-)(HCN), Me(3)SiO(-)(HCN),and PhSiF(4)(-)(HCN), many of which are otherwise difficult to generate. The bond dissociation energy of CN( )(HCN), generated by using this approach, has been measured by using energy resolved collision-induced issociation (CID) to be 0.87 +/- 0.07 eV. PMID- 16352439 TI - A high-performance liquid chromatography-tandem mass spectrometry method for quantitation of nitrogen-containing intracellular metabolites. AB - A comprehensive method of quantifying intracellular metabolite concentrations would be a valuable addition to the arsenal of tools for holistic biochemical studies. Here, we describe a step toward the development of such method: a quantitative assay for 90 nitrogen-containing cellular metabolites. The assay involves reverse-phase high-performance liquid chromatography separation followed by electrospray ionization and detection of the resulting ions using triple quadrupole mass spectrometry in selected reaction monitoring mode. For 79 of the 90 metabolites, the assay is linear with a limit of detection of 10 ng/mL or less. Using this method, 36 metabolites can be reliably detected in extracts of the bacterium Salmonella enterica, with the identity of each metabolite confirmed by the presence, on growing of the bacteria in (13)C-glucose, of a peak corresponding to the isotope-labeled form of the compound. Quantitation in biological samples is performed by mixing unlabeled test cell extract with (13)C labeled standard extract, and determining the (12)C/(13)C-ratio for each metabolite. Using this approach, the metabolomes of growing (exponential phase) and carbon-starved (stationary phase) bacteria were compared, revealing 16 metabolites that are significantly down-regulated and five metabolites that are significantly up-regulated, in stationary phase. PMID- 16352441 TI - An open design microfabricated nib-like nanoelectrospray emitter tip on a conducting silicon substrate for the application of the ionization voltage. AB - This paper describes a novel emitter tip having the shape of a nib and based on an open structure for nano-electrospray ionization mass spectrometry (nanoESI MS). The nib structure is fabricated with standard lithography techniques using SU-8, an epoxy-based negative photoresist. The tip is comprised of a reservoir, a capillary slot and a point-like feature, and is fabricated on a silicon wafer. We present here a novel scheme for interfacing such nib tips to MS by applying the ionization voltage directly onto the semi-conductor support. The silicon support is in direct contact with the liquid to be analyzed at the reservoir and microchannel level, thus allowing easy use in ESI-MS. This scheme is especially advantageous for automated analysis as the manual step of positioning a metallic wire into the reservoir is avoided. In addition, the analysis performance was enhanced compared with the former scheme, as demonstrated by the tests of standard peptides (gramicidin S, Glu-fibrinopeptide B). The limit of detection was determined to be lower than 10(-2) microM. Due to their enhanced performance, these microfabricated sources might be of great interest for analysis requiring very high sensitivity, such as proteomics analysis using nanoESI-MS. PMID- 16352442 TI - Structural analysis of kappa-carrageenan [corrected] sulfated oligosaccharides by positive mode nano-ESI-FTICR-MS and MS/MS by SORI-CID. AB - Structural analysis of sulfated oligosaccharides from kappa-carrageenan of up to ten residues (MW >2 kDa) was successfully carried out by positive mode nano-ESI FTICR-MS together with MS/MS using sustained off-resonance irradiation-collision induced dissociation (SORI-CID). Glycosidic bond cleavage reactions via the B- and Y-types of fragmentation were observed and enabled complete sequencing of the oligosaccharide samples. The positions of the labile sulfate substituents were observable using SORI-CID, enabling the determination of the sequence of the sulfated residues. PMID- 16352443 TI - Co-occurrence of boundary and resonance ejection in a multiplexed rectilinear ion trap mass spectrometer. AB - A method is reported for evaluating ion trap mass analyzers by selection of operating conditions under which both boundary and resonance ejection peaks occur in a single mass scan. The choice of frequency and amplitude of the auxiliary waveform applied for resonance ejection can be such as to produce a resonance ejection mass spectrum with unit resolution or, under selected conditions, signals attributable to both boundary and resonance ejection in a single mass scan. The contrasting mass resolution associated with these two ejection processes is evident in these data. The co-occurrence of the two ejection phenomena is ascribed to the effects of higher-order fields; it is more marked in some rectilinear ion traps (RITs) than in other nominally identical devices, leading to the possibility of using it to compare individual mass analyzers in multiplexed instruments. The method is used to compare multiple ion traps driven by the same RF signal in a fully-multiplexed mass spectrometer, composed of parallel ion source/mass analyzer/detector channels each housed in one quadrant of a specialized vacuum chamber. PMID- 16352444 TI - Electrospray mass spectral fragmentation study of N,N'-disubstituted imidazolium ionic liquids. AB - The tandem positive electrospray mass spectrometry (ESMS(n)) fragmentation of ionic liquids incorporating the 1-methyl-imidazolium ring substituted on N(II) with an alkyl chain functionalized with an alcohol, carboxylic acid, or an iodobenzyl or iodobenzoyl ester is presented for the first time. The influence of chain length and function is studied. Esterified structures led to intense CID fragments lacking the imidazolium ring allowing full characterization of the ester moiety. Fragment ion compositions for this interesting and newly important class of compounds are established through accurate mass data and deuterium labeling. The presence of the cationic ring system produces intense even electron molecular cations in electrospray that undergo multiple stages of CID to yield fragments which often are radical cations. Unusual losses of methyl and hydrogen radicals are frequently noted. PMID- 16352445 TI - Ion-exchange chromatography followed by ESI-MS for quantitative analysis of sugar monophosphates from glucose catabolism. AB - The aim of this work is to establish a quantitative method to determine the ratio of [U-(13)C] labeled to unlabeled hexose monophosphates isolated from yeast extracts. This is accomplished by anion exchange chromatography and mobile phase desalting followed by electrospray (ESI) mass spectrometry. We test the method with the analysis of a sample of biological origin. Previously developed analytical techniques are not adequate to accomplish mass spectrometric analysis of these and other small monosaccharide systems because of interference from salt clusters. By lowering the ionic strength of the mobile phase and using a simplified injection system to the mass spectrometer, we were able to obtain data on the relative abundance of the hexose monophosphates. PMID- 16352446 TI - A comparative study of the fragmentation of neutral lactooligosaccharides in negative-ion mode by UV-MALDI-TOF and UV-MALDI ion-trap/TOF mass spectrometry. AB - Structure analyses of underivatized neutral lacto oligosaccharides are systematically performed by ultraviolet matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (UV-MALDI TOF MS) and UV MALDI ion-trap time-of-flight mass spectrometry (ion-trap/TOF MS) acquired in negative-ion mode. Interestingly, their fragmentation significantly differ each other. In postsource decay (PSD) in UV-MALDI TOF MS, cross-ring cleavage at the reducing terminal predominates. On the other hand, glycosyl bond cleavage (C-type fragmentation) takes place preferentially in collision induced dissociation (CID) in UV-MALDI ion-trap/TOF MS. The cross-ring cleavage in PSD similar to that in in source decay occurs via a prompt reaction path characteristic of the UV-MALDI process itself. The product ion spectra of UV-MALDI ion-trap/TOF MS are similar to the electrospray ionization (ESI) ion-trap or quadrupole/TOF CID product ion spectra. During ion-trap/TOF MS experiments, the deprotonated molecular ions survive for several tens of milliseconds after CID event because the high internal energy chlorinated precursor ions are cooled by collisional cooling in the ion trap. The results obtained suggest that the PSD from the chlorinated precursor ion in UV-MALDI TOF MS might proceed as a two-step reaction; in the first, a high internal energy deprotonated molecular ion is generated as a reaction intermediate during the flight in the drift tube, and in the second, the rapid decomposition from the deprotonated molecular ion takes place. PMID- 16352447 TI - Humoral immune responses of amphioxus Branchiostoma belcheri to challenge with Escherichia coli. AB - Humoral parameters of amphioxus Branchiostoma belcheri, including lysozyme, antimicrobial activity, microbial agglutinin and haemagglutinins were measured before and after challenge with Escherichia coli. Humoral fluids from unchallenged B. belcheri had lysozyme, antimicrobial, microbial agglutinating and haemagglutinating activities, which may represent part of the baseline level of innate immunity in this organism. After challenge with E. coli, the lysozyme activity, growth-inhibiting activities against E. coli and Vibrio alginolyticus, microbial agglutinating activities against Micrococcus lysodeikticus, Bacillus subtilis and Staphylococus aureus, and haemagglutinating activities against rabbit and human A and O erythrocytes in the humoral fluids were all increased significantly. In contrast, the agglutinating activities against Vibrio harveyi and E. coli and the haemagglutinating activity against human B erythrocytes in the humoral fluids were reduced in response to E. coli challenge. It appears that the humoral fluids of B. belcheri contain components that are able to differentiate different microbes and different human blood cell types. PMID- 16352448 TI - Coexistence of focal and idiopathic generalized epilepsy in the same patient population. AB - PURPOSE: To review the clinical, electrographic, radiological, and pathological findings of patients with coexistent idiopathic generalized and partial epilepsy syndromes. METHODS: We performed a medical record review and a phone interview with all patients hospitalized to the Cleveland Clinic epilepsy monitoring unit (EMU) between 1992 and 2002 who fulfilled clinical and EEG criteria of coexistent partial and generalized epilepsy syndromes. RESULTS: Seven patients were identified. Two (29%) were men with a mean age of 26 years. Four had a history of febrile seizures. Family history was positive in five. Mean duration of the generalized epilepsy syndrome was 11 years, and of the focal epilepsy 18 years. An equal number of patients developed focal versus generalized epilepsy first. Interictal EEG activity was predominantly generalized. Four had video-EEG documentation of both types of seizures. In the rest, only focal seizures were recorded but interictal activity strongly suggested a coexistent generalized epilepsy. MRI showed hippocampal atrophy in all, and hippocampal dysplasia in three. Five patients had PET imaging, all with hypometabolism in areas corresponding to the ictal onset on EEG. Four patients underwent epilepsy surgery with good surgical outcome and pathological confirmation of hippocampal sclerosis in all. CONCLUSION: We found a 0.2% incidence of coexistent focal and primary generalized epilepsy. Febrile seizures and a positive family history were common. Good seizure control was achieved after temporal lobectomy, even when interictal generalized activity predominated. PMID- 16352449 TI - Peroxynitrite-induced oxidation and nitration products of guanine and 8 oxoguanine: structures and mechanisms of product formation. AB - Peroxynitrite induces DNA base damage predominantly at guanine (G) and 8 oxoguanine (8-oxoG) nucleobases via oxidation reactions. Nitration products are also observed, consistent with the generation of radical intermediates that can recombine with the (.)NO(2) formed during peroxynitrite degradation. The neutral G radical, G(.), reacts with (.)NO(2) to yield 8-nitroguanine (8-nitroG) and 5 nitro-4-guanidinohydantoin (NI), while for 8-oxoG we have proposed a reactive guanidinylidene radical intermediate. The products generated during peroxynitrite mediated 8-oxoG oxidation depend on oxidant flux, with dehydroguanidinohydantoin (DGh), 2,4,6-trioxo-[1,3,5]triazinane-1-carboxamidine (CAC) and NO(2)-DGh predominating at high fluxes and spiroiminodihydantoin (Sp), guanidinohydantoin (Gh) and 4-hydroxy-2,5-dioxo-imidazolidine-4-carboxylic acid (HICA) predominating at low fluxes. Both product sets are observed at intermediate fluxes. It is therefore important in model systems to ensure that the relative concentrations are well controlled to minimize competing reactions that may not be relevant in vivo. Increasingly sophisticated systems for modeling peroxynitrite production in vivo are being developed and these should help with predicting the products most likely to be formed in vivo. Together with the emerging information on the genotoxic and mutational characteristics of the individual oxidation products, it may be found that the extent of tissue damage, mutational spectra and, hence, cancer risk may change as a function of peroxynitrite fluxes as different product combinations predominate. PMID- 16352450 TI - The effect of seawater composition and osmolality on hemolymph levels of methyl farnesoate in the green crab Carcinus maenas. AB - Green crabs, Carcinus maenas, exposed to dilute seawater (e.g., 5 ppt salinity, approximately 150 mOsm/kg) have hemolymph levels of methyl farnesoate (MF) that are up to 10-fold higher than animals in isosmotic seawater (27 ppt, approximately 800 mOsm/kg). In this paper, we examine aspects of osmotic and ionic stress to identify factors involved in elevating MF levels. MF levels did not rise after exposure to concentrated seawater, so only hypoosmotic stress elevates MF. MF levels rose in animals exposed to dilute seawater containing mannitol to make it isosmotic, indicating that the hypoosmotic rise in MF is due to decreased ion concentrations. Individual ions were investigated by exposing crabs either to isosmotic seawater with low concentrations of an ion or to dilute seawater with high concentrations of an ion. Ca(2+) and Mg(2+) in combination affected MF levels. Finally, we found that the increase in MF levels was accelerated when hemolymph osmolality was precociously lowered by partially replacing hemolymph with deionized water prior to transferring animals to dilute seawater. Thus, the 6-8 h delay between exposing crabs to dilute sea water and observing an increase in MF appears to reflect the time needed for specific hemolymph ions to decrease below a threshold concentration. PMID- 16352451 TI - Production, characterization and applications of mouse anti-grass carp (Ctenopharyngodon idellus) growth hormone monoclonal antibodies. AB - Mouse anti-grass carp growth hormone (gcGH) monoclonal antibody (MAb) secretors were produced by PEG-mediated fusion of NS-1 myeloma cells and splenic B lymphocytes of gcGH hyper-immunized mice. Positive secretors were screened by direct ELISA and cloned by limiting dilution. Three positive secretors, 21D3, 22G5 and 23B3, were obtained in a single fusion trial. Anti-gcGH MAbs were produced by growing hybridomas in the peritoneal cavity of pristane-primed mouse. The three MAbs were isotyped to be IgG2a, IgG2b and IgM, respectively. IgG MAbs were purified from ascitic fluid by Hitrap protein G column and IgM MAb was purified by gel filtration chromatography. The purified MAbs were highly specific and had moderate binding affinity. The MAbs were successfully used for the purification of native gcGH from mature grass carp pituitary extract by one-step immunoaffinity chromatography, for the quantification of gcGH by competitive sandwich ELISA, and for the probing of somatotropes in grass carp pituitary by immunohistochemistry. PMID- 16352452 TI - Common HEXB polymorphisms reduce serum HexA and HexB enzymatic activities, potentially masking Tay-Sachs disease carrier identification. AB - A DNA-proven Tay-Sachs disease (TSD) carrier and his brother were found to have serum percent Hexosaminidase A (%HexA) enzymatic activities in the non-carrier range, while the leukocyte %HexA profiles clearly identified them as TSD heterozygotes. Both their serum HexA and HexB enzymatic activities were below reference range, suggesting inheritance of mutations in both the HEXA (alpha subunit) and HEXB (beta-subunit) genes. DNA sequencing revealed that both individuals, carried the common HEXA 1277_1278insTATC mutation, and two common HEXB polymorphisms: [619A>G (+) delTG]. To determine if these HEXB polymorphisms reduce HexA and HexB enzymatic activities, 69 DNA samples from subjects previously screened enzymatically in both serum and leukocytes for TSD carrier status were selected for either high, mid-range or low serum Total Hex (defined as the sum of HexA and HexB) activities and were tested for the HEXB mutations. Further, three additional TSD carriers ascertained by the atypical pattern of normal serum %HexA but carrier leukocyte %HexA, were found to have the [delTG (+) 619A>G] genotype. In addition, the frequency of the [delTG (+) 619A>G] genotype was significantly higher (P < 0.01) in subjects with low serum HexB enzymatic activities. Given the high frequency of the [delTG (+) 619A>G] haplotype in the Ashkenazi Jewish population (approximately 10%), up to 10% of TSD carriers may have normal serum %HexA values with low total Hex. Accordingly, serum %HexA should not be the sole criterion used for carrier status determination. Where total Hex activity is reduced, further testing with leukocyte Hex profiles is indicated. PMID- 16352453 TI - Genotype-phenotype relationships involving hypertrophic cardiomyopathy-associated mutations in titin, muscle LIM protein, and telethonin. AB - BACKGROUND: TTN-encoded titin, CSRP3-encoded muscle LIM protein, and TCAP-encoded telethonin are Z-disc proteins essential for the structural organization of the cardiac sarcomere and the cardiomyocyte's stretch sensor. All three genes have been established as cardiomyopathy-associated genes for both dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM). Here, we sought to characterize the frequency, spectrum, and phenotype associated with HCM associated mutations in these three genes in a large cohort of unrelated patients evaluated at a single tertiary outpatient center. METHODS: DNA was obtained from 389 patients with HCM (215 male, left ventricular wall thickness of 21.6+/-6 mm) and analyzed for mutations involving all translated exons of CSRP3 and TCAP and targeted HCM-associated exons (2, 3, 4, and 14) of TTN using polymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC), and direct DNA sequencing. Clinical data were extracted from patient records and maintained independent of the genotype. RESULTS: Overall, 16 patients (4.1%) harbored a Z-disc mutation: 12 had a MLP mutation and 4 patients a TCAP mutation. No TTN mutations were detected. Seven patients were also found to have a concomitant myofilament mutation. Seven patients with a MLP-mutation were found to harbor the DCM-associated, functionally characterized W4R mutation. W4R-MLP was also noted in a single white control subject. Patients with MLP/TCAP associated HCM clinically mimicked myofilament-HCM. CONCLUSIONS: Approximately 4.1% of unrelated patients had HCM-associated MLP or TCAP mutations. MLP/TCAP-HCM phenotypically mirrors myofilament-HCM and is more severe than the subset of patients who still remain without a disease-causing mutation. The precise role of W4R-MLP in the pathogenesis of either DCM or HCM warrants further investigation. PMID- 16352455 TI - Ultrasonic degradation of oxalic acid in aqueous solutions. AB - This paper describes the ultrasonic degradation of oxalic acid. The effects of ultrasonic power, H(2)O(2), NaCl, external gases on the degradation of oxalic acid were investigated. Reactor flask containing oxalic acid was immersed in the ultrasonic bath with water as the coupling fluid. Representative samples withdrawn were analysed by volumetric titration. Degradation degree of oxalic acid increased with increasing ultrasonic power. It was observed that H(2)O(2) has negative contribution on the degradation of oxalic acid and there was an optimum concentration of NaCl for enhancing the degradation degree of oxalic acid. Although bubbling nitrogen gave higher degradation than that for bubbling air, both gases (for 20 min before sonication and during sonication together) could not help to enhance the degradation of oxalic acid when compared with the degradation without gas passage. PMID- 16352454 TI - Validation of a heparan sulfate-derived disaccharide as a marker of accumulation in murine mucopolysaccharidosis type IIIA. AB - Mucopolysaccharidosis type IIIA (MPS IIIA) is a neurodegenerative lysosomal storage disorder resulting from sulfamidase deficiency, which leads to accumulation of heparan sulfate within lysosomes. We have determined the time course of accumulation of a disaccharide [hexosamine-N-sulfate[alpha 1,4]hexuronic acid; HNS-UA] marker of heparan sulfate storage within the brain, liver, and spleen of a naturally occurring mouse model of MPS IIIA. HNS-UA is detectable in the brain of affected mice on the day of birth, when it is significantly increased compared to normal control mice. As mice age, this compound steadily accumulates until a plateau is reached at approximately 20 weeks. A similar rate of accumulation of HNS-UA is seen in the liver and spleen of affected mice. Intracerebral delivery of recombinant human sulfamidase reduced the amount of HNS-UA present in segments of the brain receiving the correcting enzyme, thus demonstrating the effectiveness of enzyme replacement therapy within the central nervous system of affected mice. This finding therefore provides evidence for the use of the disaccharide HNS-UA to monitor the effect of therapies for this condition in humans, when treatment strategies are devised. PMID- 16352456 TI - Systemic lupus-erythematosus: deoxyribonuclease 1 in necrotic chromatin disposal. AB - Systemic lupus-erythematosus is an auto-immune-disease characterized by pathogenic anti-nuclear auto-antibodies. These form immune-complexes that after deposition at basal membranes at various locations initiate inflammatory reactions. There is a clear genetic and gender predisposition (females are affected 10 times more frequently), but also infectious agents and further environmental factors have been shown to be causative for the initiation of the disease. It has been suggested that the auto-antibodies arise after release and/or inefficient removal of nuclear components during cell death (defective cellular "waste disposal" theory). So far, increased apoptotic cell death has been made responsible, but recent data suggest that defective cellular waste disposal during/after necrosis may also lead to the release and prolonged exposure of nuclear components. Here, we concentrate on chromatin disposal during necrosis and the involvement of Deoxyribonuclease 1 in this process with respect to its possible role in the prevention of anti-nuclear auto-immunity. PMID- 16352457 TI - Green tea polyphenol inhibits Mycobacterium tuberculosis survival within human macrophages. AB - Lack of maturation of phagosomes containing pathogenic Mycobacterium tuberculosis within macrophages has been widely recognized as a crucial factor for the persistence of mycobacterial pathogen. Host molecule tryptophan-aspartate containing coat protein (TACO) has been shown to play a crucial role in the arrest of such a maturation process. The present study was addressed to understand whether or not polyphenols derived from green tea could down-regulate TACO gene transcription. And if yes, what impact TACO gene down-regulation has on the uptake/survival of M. tuberculosis within macrophages. The reverse transcriptase polymerase chain reaction and reporter assay technology, employed in this study, revealed that the major component of green tea polyphenols, epigallocatechin-3-gallate had the inherent capacity to down-regulate TACO gene transcription within human macrophages through its ability to inhibit Sp1 transcription factor. We also found out that TACO gene promoter does contain Sp1 binding sequence using bioinformatics tools. The down-regulation of TACO gene expression by epigallocatechin-3-gallate was accompanied by inhibition of mycobacterium survival within macrophages as assessed through flow cytometry and colony counts. Based on these results, we propose that epigallocatechin-3-gallate may be of importance in the prevention of tuberculosis infection. PMID- 16352458 TI - Understanding the cytochrome bc complexes by what they don't do. The Q-cycle at 30. AB - The cytochrome (cyt) bc(1), b(6)f and related complexes are central components of the respiratory and photosynthetic electron transport chains. These complexes carry out an extraordinary sequence of electron and proton transfer reactions that conserve redox energy in the form of a trans-membrane proton motive force for use in synthesizing ATP and other processes. Thirty years ago, Peter Mitchell proposed a general turnover mechanism for these complexes, which he called the Q cycle. Since that time, many opposing schemes have challenged the Q-cycle but, with the accumulation of large amounts of biochemical, kinetic, thermodynamic and high-resolution structural data, the Q-cycle has triumphed as the accepted model, although some of the intermediate steps are poorly understood and still controversial. One of the major research questions concerning the cyt bc(1) and b(6)f complexes is how these enzymes suppress deleterious and dissipative side reactions. In particular, most Q-cycle models involve reactive semiquinone radical intermediates that can reduce O(2) to superoxide and lead to cellular oxidative stress. Current models to explain the avoidance of side reactions involve unprecedented or unusual enzyme mechanisms, the testing of which will involve new theoretical and experimental approaches. PMID- 16352459 TI - Transmission dynamics of Taenia solium and potential for pig-to-pig transmission. AB - Taenia solium taeniasis/cysticercosis is one of few potentially eradicable infectious diseases and is the target of control programs in several countries. The larval stage of this zoonotic cestode invades the human brain and is responsible for most cases of adult-onset epilepsy in the world. Our current understanding of the life cycle implicates humans as the only definitive host and tapeworm carrier, and thus the sole source of infective eggs that are responsible for cysticercosis in both human and pigs through oral-faecal transmission. Here we review transmission dynamics of porcine cysticercosis including an alternative pig-to-pig route of transmission, previously not suspected to exist. Second-hand transmission of T. solium eggs could explain the overdispersed pattern of porcine cysticercosis, with few pigs harbouring heavy parasite burdens and many more harbouring small numbers of parasites. PMID- 16352460 TI - Molecular survival strategies of Echinococcus multilocularis in the murine host. AB - Larval infection with Echinococcus multilocularis starts with the intrahepatic postoncospheral development of a metacestode that-at its mature stage-consists of an inner germinal and an outer laminated layer (GL & LL). In certain cases, an appropriate host immune response may inhibit parasite proliferation. Several lines of evidence obtained in vivo and in vitro indicate the important bio protective role of the LL. For instance, the LL has been proposed to protect the GL from nitric oxide produced by periparasitic macrophages and dendritic cells, and also to prevent immune recognition by surrounding T cells. On the other hand, the high periparasitic NO production by peritoneal exsudate cells contributes to periparasitic immunosuppression, explaining why iNOS deficienct mice exhibit a significantly lower susceptibility towards experimental infection. The intense periparasitic granulomatous infiltration indicates a strong host-parasite interaction, and the involvement of cellular immunity in control of the metacestode growth kinetics is strongly suggested by experiments carried out in T cell deficient mouse strains. Carbohydrate components of the LL, such as Em2(G11) and Em492, as well as other parasite metabolites yield immunomodulatory effects that allow the parasite to survive in the host. I.e., the IgG response to the Em2(G11)-antigen takes place independently of alpha-beta+CD4+T cells, and in the absence of interactions between CD40 and CD40 ligand. Such parasite molecules also interfere with antigen presentation and cell activation, leading to a mixed Th1/Th2-type response at the later stage of infection. Furthermore, Em492 and other (not yet published) purified parasite metabolites suppress ConA and antigen stimulated splenocyte proliferation. Infected mouse macrophages (AE-MO) as antigen presenting cells (APC) exhibited a reduced ability to present a conventional antigen (chicken ovalbumin, C-Ova) to specific responder lymph node T cells when compared to normal MO. As AE-MO fully maintain their capacity to appropriately process antigens, a failure in T cell receptor occupancy by antigen Ia complex or/and altered co-stimulatory signals can be excluded. Studying the status of accessory molecules implicated in T cell stimulation by MO, it could be shown that B7-1 (CD80) and B7-2 (CD86) remained unchanged, whereas CD40 was down regulated and CD54 (=ICAM-1) slightly up-regulated. FACS analysis of peritoneal cells revealed a decrease in the percentage of CD4+ and CD8+T cells in AE infected mice. Taken together the obstructed presenting-activity of AE-MO appeared to trigger an unresponsiveness of T cells leading to the suppression of their clonal expansion during the chronic phase of AE infection. Interesting information on the parasite survival strategy and potential can be obtained upon in vitro and in vivo treatment. Hence, we provided very innovative results by showing that nitazoxanide, and now also, respectively, new modified compounds may represent a useful alternative to albendazole. In the context of chemotherapeutical repression of parasite growth, we searched also for parasite molecules, whose expression levels correlate with the viability and growth activity of E. multilocularis metacestode. Expression levels of 14-3-3 and II/3 10, relatively quantified by realtime reverse transcription-PCR using a housekeeping gene beta-actin, were studied in permissive nu/nu and in low permissive wild type BALB/c mice. At 2 months p.i., the transcription level of 14 3-3 was significantly higher in parasites actively proliferating in nu/nu mice compared to parasites moderately growing in wild type mice. Immunoblotting experiments confirmed at the protein level that 14-3-3 was over-expressed in parasites derived from nu/nu mice at 2 months p.i. In vitro-treatment of E. multilocularis with an anti-echinococcal drug nitazoxanide for a period of 8 days resulted in a significant decrease of both 14-3-3 and II/3-10 transcription levels, which correlated with the kinetics of a housekeeping gene, beta-actin. This indicates that 14-3-3-exhibits a good potential as a molecular marker to assess viability and growth activity of the parasite. PMID- 16352461 TI - Recombinant antigens for serodiagnosis of cysticercosis and echinococcosis. AB - Diagnosis of cysticercosis/echinococcosis is primarily based on imaging techniques. These imaging techniques are sometimes limited by the small size of visualized lesions and atypical images, which are difficult to be distinguished from abscesses or neoplasms. Therefore, efforts have been directed toward identification and characterization of specific antigens of parasites for development of serodiagnostic method that can detect specific antibody. For cysticercosis, glycoproteins of 10-26 kDa in cyst fluid of Taenia solium have been widely accepted for serodiagnosis purpose. The glycoproteins consist of a very closely related family of 8-kDa proteins. We identified four genes (designated Ag1, Ag1V1, Ag2 and Ag2V1) encoding the 7- and 10-kDa polypeptides. Based on the antigenicities of these clones, Ag1V1 and Ag2 were chosen as ELISA antigens and the Ag1V1/Ag2 chimeric protein was expressed. The Ag1V1/Ag2 chimeric protein showed the similar sensitivity and specificity as the native glycoproteins. For alveolar echinococcosis, the 65-kDa protein of Echinococcus multilocularis protoscolices and Em18 has been considered as serodiagnostic antigens. The sensitivity and specificity of Em18 are very compatible to those of the recombinant 65-kDa protein. Recently, we demonstrated that Em18 was the proteolytic product of the 65-kDa protein following the action by cysteine proteinases. From the information of N-terminal amino acid sequences, molecular size and isoelectric point of Em18, recombinant Em18 ((349)K to (508)K of the 65 kDa protein, RecEm18) was expressed and evaluated for serodiagnostic value. RecEm18 has the potential for use in the differential serodiagnosis of alveolar echinococcosis. PMID- 16352462 TI - Echinococcus granulosus in Australia, widespread and doing well! AB - Echinococcus granulosus is the only member of the Genus Echinococcus to occur in Australia. The major biomass of E. granulosus occurs in wildlife. The wildlife transmission cycle is predominantly perpetuated via a predator/prey interaction between wild dogs (dingoes and dingo/domestic dog hybrids) and macropodid marsupials (wallabies and kangaroos). Other wildlife hosts include foxes, wombats and feral pigs. This wildlife reservoir for E. granulosus "spills over" to help maintain a domestic cycle through E. granulosus-infected wild dogs defecating on pasture, transmitting infection to livestock and some farmers and hunters feeding hydatid-infected offal of macropodids or feral pigs to domestic dogs. The potential transmission risk to humans using public picnic and camping areas in parks and forests, especially in the southeastern Australia, could be substantially reduced through regular distribution of baits containing praziquantel. Encroachment of wild dogs and foxes into urban centers presents a new potential path of transmission from wildlife to humans. PMID- 16352463 TI - Laboratory animal models for human Taenia solium. AB - Human beings are the only hosts of adult Taenia solium; thus, many aspects of the host-parasite relationship are unknown. The development of successful experimental models of taeniasis allows in-depth investigations of the host parasite relationship. We established experimental models in hamsters, gerbils and chinchillas. Here we review our findings regarding the characteristics of the tapeworms, their anchoring site and development, as well as the humoral and cellular immune response they elicit. We also used statistics to analyze the data obtained in different infections performed along several years. Furthermore, we compared the size of T. solium rostellum and strobila recovered from hamsters and gerbils to those obtained from humans. Our data indicate that these rodents are adequate experimental models for studying T. solium in its adult stage; that parasites induce immune responses and that hamsters seem to be more permissive hosts than gerbils, since parasites survive for longer times, grow longer and develop more, and the inflammatory response in the intestinal mucosa against T. solium is moderate. Finally, chinchillas are the most successful experimental definitive model for adult T. solium, since tapeworms with gravid proglottids are obtained, and the life cycle can be continued to the intermediate host. PMID- 16352464 TI - Genetic variation in Taenia solium. AB - Neurocysticercosis is a major zoonotic larval cestode infection that has a worldwide distribution and is of significant public health importance. Knowledge of the genetic structure of Taenia solium can be applied to the epidemiology and transmission of this disease, since genetic variants may differ in infectivity and pathogenicity. Molecular epidemiological approaches can also enable detailed studies of transmission. On a global scale, mitochondrial markers have differentiated between T. solium isolates from Asia and isolates from Africa/Latin America. Intraspecific variation in T. solium has been detected to some extent, using RAPD markers to differentiate between T. solium populations from different regions within Mexico. Markers currently available for T. solium have not been used to analyse genetic variation at the community or local level. The development of highly polymorphic markers such as microsatellites in T. solium can provide the means to examine genetic heterogeneity of tapeworm infection at the household, community and regional level. Preliminary studies suggest it is possible to analyse population genetic variation in communities using a range of polymorphic markers. PMID- 16352465 TI - The present situation of echinococcosis in Europe. AB - The taxonomy of Echinococcus is presently undergoing major changes, the paraphyletic Echinococcus granulosus being split into several distinct species. In this review, an attempt is made to assess the present epidemiological situation in Europe separately for each species (Echinococcus multilocularis, Echinococcus granulosus sensu stricto, Echinococcus equinus, Echinococcus ortleppi, and Echinococcus sp.). For E. multilocularis, an increasing density of infected host animals is apparent in central Europe, and, possibly, a range increase has occurred. Prevalence rates in foxes have risen in many agriculturally dominated landscapes of France, The Netherlands, Germany, Austria, Slovakia and Poland, but the lifecycle is now also established in many urban areas, where red foxes occur with high population densities. E. granulosus s. s. (the former 'sheep strain') is still frequent and a public health problem in many parts of the Mediterranean region and re-emergence after failed control campaigns is observed or suspected in Bulgaria and Wales. No recent data on the cattle transmitted E. ortleppi and the horse-transmitted E. equinus are available, but their relevance for human health seems to be minor. The same may apply to the 'pig strain' and the newly described 'European cervid strain', which both belong to a cluster of genotypes whose taxonomy is not yet resolved (Echinococcus sp.). PMID- 16352466 TI - Solvent effects on infrared spectroscopy of 17beta-hydroxy-17-methyl-2-oxa-5alpha androstan-3-one in single solvents. AB - Infrared spectroscopy studies of 17beta-hydroxy-17-methyl-2-oxa-5alpha-androstan 3-one (HMOA) in 20 different pure organic solvents were undertaken to investigate the solute-solvent interactions and to correlate solvent properties, such as the KBM equation, the solvent acceptor number (AN), Schleyer's linear free energy equation (G-value), Dimroth and Reichardt's parameter E(T)(30) and Brownstein's parameter S with the frequency shifts of the carbonyl stretching vibration for solute. The result indicated that the KBM relationship is unsuitable to the complicated molecules. These frequencies exhibited a better correlation with the solvent parameters AN and G-value than others. PMID- 16352467 TI - Suppression of mast cell degranulation by a novel ceramide kinase inhibitor, the F-12509A olefin isomer K1. AB - Antigen-induced degranulation of mast cells plays a pivotal role in allergic and inflammatory responses. Recently, ceramide kinase (CERK) and its phosphorylated product ceramide 1-phosphate (C1P) have emerged as important players in mast cell degranulation. Here, we describe the synthesis of a novel F-12509A olefin isomer, K1, as an effective CERK inhibitor. In vitro kinase assays demonstrated that K1 effectively inhibits CERK without inhibiting sphingosine kinase and diacylglycerol kinase. Treating RBL-2H3 cells with K1 reduced cellular C1P levels to 40% yet had no effect on cell growth. Furthermore, treatment with K1 significantly suppressed both calcium ionophore- and IgE/antigen-induced degranulation, indicating that K1 interferes with signals that happen downstream of Ca(2+) mobilization. Finally, we show that K1 affects neither IgE/antigen induced global tyrosine phosphorylation nor subsequent Ca(2+) elevation, suggesting a specificity for CERK-mediated signals. Our novel CERK inhibitor provides a useful tool for studying the biological functions of CERK and C1P. Moreover, to our knowledge, this is the first report demonstrating that inhibition of CERK suppresses IgE/antigen-induced mast cell degranulation. This finding suggests that CERK inhibitors might be a potential therapeutic tool in the treatment of allergic diseases. PMID- 16352468 TI - Phylogeny of marine Gregarines (Apicomplexa)--Pterospora, Lithocystis and Lankesteria--and the origin(s) of coelomic parasitism. AB - Gregarines constitute a large group of apicomplexans with diverse modes of nutrition and locomotion that are associated with different host compartments (e.g. intestinal lumena and coelomic cavities). A broad molecular phylogenetic framework for gregarines is needed to infer the early evolutionary history of apicomplexans as a whole and the evolutionary relationships between the diverse ultrastructural and behavioral characteristics found in intestinal and coelomic gregarines. To this end, we sequenced the SSU rRNA gene from (1) Lankesteria abbotti from the intestines of two Pacific appendicularians, (2) Pterospora schizosoma from the coelom of a Pacific maldanid polychaete, (3) Pterospora floridiensis from the coelom of a Gulf Atlantic maldanid polychaete and (4) Lithocystis sp. from the coelom of a Pacific heart urchin. Molecular phylogenetic analyses including the new sequences demonstrated that several environmental and misattributed sequences are derived from gregarines. The analyses also demonstrated a clade of environmental sequences that was affiliated with gregarines, but as yet none of the constituent organisms have been described at the ultrastructural level (apicomplexan clade I). Lankesteria spp. (intestinal parasites of appendicularians) grouped closely with other marine intestinal eugregarines, particularly Lecudina tuzetae, from polychaetes. The sequences from all three coelomic gregarines branched within a larger clade of intestinal eugregarines and were similarly highly divergent. A close relationship between Pterospora schizosoma (Pacific) and Pterospora floridiensis (Gulf Atlantic) was strongly supported by the data. Lithocystis sp. was more closely related to a clade of marine intestinal gregarines consisting of Lankesteria spp. and Lecudina spp. than it was to the Pterospora clade. These data suggested that coelomic parasitism evolved more than once from different marine intestinal eugregarines, although a larger taxon sample is needed to further explore this inference. PMID- 16352469 TI - Safety of ESAT-6. AB - A recombinant dimer of the Mycobacterium tuberculosis (MTb) 6 kDa early secreted antigenic target (ESAT-6) was produced in Lactococcus lactis. Pharmacodynamic and safety studies were carried out in guinea pigs, rats, mice and dogs with intradermal (id), subcutaneous (sc) and intravenous (iv) administration of the antigen. In contrast to tuberculin purified protein derivative (PPD) the recombinant dimer (rdESAT-6) was able to discriminate MTb infection from BCG vaccination in vivo. In guinea pigs sensitized by infection with MTb, 1 microg rdESAT-6 gave a mean delayed-type hypersensitivity (DTH) response of 22 mm, a significantly stronger reaction than in animals sensitised by the environmental mycobacteria M. kansasii, M. szulgai and M. marinum. rdESAT-6 proved to be a safe tuberculin reagent in a dose range of 1-1000 microg with no or only minor local reactions. PMID- 16352470 TI - Serum anti-mycobacterial heat shock proteins antibodies in sarcoidosis and tuberculosis. AB - Sarcoidosis (SA) is a multisystem granulomatous disorder of unknown etiology. Infectious organisms, e.g. Mycobacterium tuberculosis, genetic factors and autoimmunity are considered as etiologic agents. Pathologic similarities between SA and tuberculosis (TB) suggest M. tuberculosis heat shock proteins (Mtb-hsp) as causative factors. To test the "mycobacterial" origin of SA, we evaluated the presence of serum anti-Mtb-hsp70, -Mtb-hsp65 and -Mtb-hsp16 antibodies in SA and TB. Analysis of anti-Mtb-hsp antibodies was carried out in 37 patients with SA, 29 patients with TB and 18 healthy individuals by ELISA. Our results show a significantly higher occurrence of anti-Mtb-hsp70 antibodies in SA and TB patients than in controls. The anti-Mtb-hsp65 and -Mtb-hsp16 antibodies occured significantly more often in TB than in controls and SA. We found significantly higher percentages of anti-Mtb-hsp70 and -Mtb-hsp65 antibodies in Stage II compared to Stage I of SA. Analysis of anti-Mtb-hsp antibodies levels revealed significantly higher anti-Mtb-hsp70 antibodies level in SA and TB than in controls. Significantly higher frequency and levels of anti-Mtb-hsp70 than anti Mtb-hsp65 and -Mtb-hsp16 antibodies was found in SA only. In summary, the frequency and level of anti-Mtb-hsp70 antibodies were comparable between SA and TB but were significantly higher compared to controls and other tested Mtb-hsp. These data may suggest a role for Mtb-hsp70 protein in the pathogenesis of sarcoidosis. PMID- 16352471 TI - Excessive daytime sleepiness and sleep complaints among children with epilepsy. AB - OBJECTIVE: Excessive daytime sleepiness (EDS) and sleep complaints are common among adults with epilepsy. We hypothesized that children with epilepsy have worse daytime sleepiness compared with controls. METHODS: Children with and without epilepsy between ages 8 and 18 were recruited for the study. Parents and children were asked to fill out the Pediatric Sleep Questionnaire (PSQ) and Pediatric Daytime Sleepiness Scale (PDSS), respectively. The Mann-Whitney U test was used for group comparisons, with the Fischer exact or chi2 test for categorical variables. Regression analysis was used to identify predictors of EDS. RESULTS: Twenty-six patients and matched controls were recruited for the study. Parents of children with epilepsy more often reported EDS (P < 0.001), symptoms of sleep-disordered breathing (P < 0.001), and parasomnias (P < 0.001) compared with controls. On the PDSS, children with epilepsy reported worse daytime sleepiness scores compared with controls (15.48 +/- 6.4 vs 11.88 +/- 5.25, P = 0.037). Based on conditional logistic regression modeling, symptoms of excessive daytime sleepiness [corrected] (OR = 15.3, 95% CI = 1.4-166.6) and parasomnias (OR = 12.4, 95% CI = 1.01-151.6) were significantly associated with having epilepsy when adjusted for duration of nightime sleep. Further, 10 children (38.5%) with epilepsy reported positive sleep-disordered breathing, whereas no one in the control group reported SDB (P < 0.001) [corrected] Epilepsy syndrome, anticonvulsants used, and presence or absence of seizure freedom, however, were not significant predictors of EDS among patients. CONCLUSIONS: Daytime sleepiness appears to be common in children with epilepsy, and may be due to underlying sleep disorders. Further confirmatory studies are needed using screening questionnaires and formal sleep studies to systematically study the prevalence of sleep complaints and role of sleep disorders in these patients. PMID- 16352472 TI - Respiratory and body temperature modulation by adenosine A1 receptors in the anteroventral preoptic region during normoxia and hypoxia. AB - The present study was undertaken to investigate adenosine as a simultaneous mediator of hypoxia-induced hyperventilation and regulated hypothermia in the anteroventral preoptic region (AVPO), the thermointegrative region of the central nervous system (CNS). Accordingly, we predicted that injection of aminophylline and DPCPX, non-selective and A(1) receptor antagonists, respectively, into the AVPO would exacerbate the ventilatory response and lessen the drop in body temperature (T(b)) caused by hypoxia. We measured ventilation (V ) and T(b) of conscious Wistar rats before and after AVPO injection of aminophylline (1 and 10 microg/100 nL) or DPCPX (17.5 and 175 ng/100 nL), or their respective vehicles, followed by 30 min of hypoxia (7% O(2)). Vehicles and the lower doses of both antagonists had no effect on V and T(b) during normoxia or hypoxia. The higher doses of aminophylline and DPCPX increased (P<0.05) the hypoxia-induced hyperventilation, whereas the drop in T(b) elicited by hypoxia was attenuated (P<00.05) by DPCPX only. This higher DPCPX dose also increased T(b) during normoxia. The present data is consistent with the notion that adenosine plays an inhibitory role in respiratory and metabolic regulation, in a way that A(1) receptors stimulation in the AVPO inhibits ventilatory drive during hypoxia and tonically modulates basal T(b). PMID- 16352473 TI - Cochrane reviews in neonatology: past, present and future. AB - The Neonatal Review Group of the Cochrane Collaboration is dedicated to improving outcomes of newborn infants through the collection and synthesis of the highest quality evidence. Much has been achieved with limited resources. Future challenges for the group are to maintain and extend current reviews of therapeutic interventions, to develop bridging reviews to assist clinicians in applying current evidence more easily, to expand the scope of the Cochrane Library to include diagnostic tests, and to utilize techniques such as prospective meta-analysis to answer remaining questions in the field. In future, the Neonatal Review Group needs to assist reviewers in developing countries to prepare reviews relevant to their settings that will reduce the global burden of neonatal mortality and morbidity. PMID- 16352474 TI - The mouse's eye and Mfrp: not quite human. PMID- 16352475 TI - Mutation screen of the membrane-type frizzled-related protein (MFRP) gene in patients with inherited retinal degenerations. AB - MFRP is a member of the frizzled-related protein family and contains a cysteine rich domain essential for Wnt binding and signaling. MFRP is highly expressed in the retinal pigment epithelial cells of the eye. A splice donor mutation in the mouse ortholog of Mfrp is responsible for photoreceptor degeneration in the rd6 mouse. For these reasons, we investigated MFRP as a candidate gene for a phenotype associated with mutations. We screened 152 patients with inherited retinal degenerations including retinitis pigmentosa, Leber congenital amaurosis and Stargardt macular dystrophy. We identified five polymorphisms in the 5' untranslated region, four missense changes, six isocoding variants and four intronic changes. None of the sequence variants were interpreted as pathogenic. PMID- 16352476 TI - Unilateral isolated microphthalmia inherited as an autosomal recessive trait. AB - PURPOSE: To report a family with unilateral isolated microphthalmia showing an autosomal recessive pattern of inheritance. CASE REPORT: We report a family in which three out of four children, one male and monozygotic female twins, were born with unilateral isolated microphthalmia to healthy consanguineous parents. One twin additionally had a horseshoe kidney. Rare cases of familial isolated microphthalmia/anophthalmia have been previously described. This is the first report of a family with autosomal recessive isolated microphthalmia occurring unilaterally in all affected individuals. It remains unknown how this inherited genetic disease results in unilateral manifestation. CONCLUSION: Mirror imaging of this condition in the monozygotic twins may help elucidate the underlying mechanism. The constellation of features in this family may contribute to solve remaining questions of research into symmetry and asymmetry. PMID- 16352477 TI - Meesmann corneal dystrophy (MECD): report of 2 families and a novel mutation in the cornea specific keratin 12 (KRT12) gene. AB - PURPOSE: Meesmann corneal dystrophy (MECD) is an autosomal dominant disorder affecting the corneal epithelium. It is caused by heterozygous mutations in KRT3 or KRT12 gene. Actually, 14 mutations have been reported, 1 in KRT3 and 13 in KRT12. These genes were screened in several patients suffering from MECD. METHODS: Patients from 2 families were screened for mutation in KRT3 and KRT12. Exons were PCR-amplified and directly sequenced. The new mutation was checked by DHPLC in 51 control individuals of Swiss origin. RESULTS/CONCLUSIONS: In one family, the M129T heterozygous mutation was observed in KRT12. In the second family, we identified a novel I426S heterozygous mutation in exon 6 of KRT12. PMID- 16352478 TI - RPGRIP1 is mutated in Leber congenital amaurosis: a mini-review. AB - RPGRIP1 encodes the retinitis pigmentosa GTPase interacting protein 1 and interacts with RPGR, the latter represents the major X-linked RP (XRRP) gene, as it accounts for 70-80% of the XRRP patients and up to 13% of all RP patients. RPGRIP1 contains a C-terminal RPGR interacting domain (RID) and a coiled-coil (CC) domain, which is homologous to proteins involved in vesicular trafficking. The interactions between the two proteins is between the RCC1-homologous domain of RPGR (RHD) and the RPGR-interacting domain of RPGRIP1 (RID). Both proteins co localize to the photoreceptor connecting cilium and RPGRIP1 appears to be a structural component of the ciliary axoneme of the connecting cilium (which connects the inner to the outer segment of the photoreceptors) of both rods and cones and functions to anchor RPGR within the cilium.RPGRIP1 loci encode several different isoforms, which have distinct cellular, sub cellular and biochemical properties. RPGRIP1 is uniquely expressed in amacrine cells of the inner retina. Knockout mice studies have shown that RPGRIP1 is required for disc morphogenesis of the outer segments in the mouse, perhaps by regulating cytoskeleton dynamics. Thus far RPGRIP1 appears to be only mutated in LCA and is associated with 6% of LCA in two series. The purpose of this review is to highlight recent advances in our understanding of RPGRIP1 function in normal and diseased retinas. PMID- 16352479 TI - Coats-type retinal telangiectasia in case of Kabuki make-up syndrome (Niikawa Kuroki syndrome). AB - PURPOSE: To report a case of Kabuki make-up syndrome (KMS) or Niikawa-Kuroki syndrome with Coats-type retinal telangiectasia, which has not been previously reported. METHODS: Observational case report. CONCLUSIONS: There have been only two reports of retinal pigmentation abnormalities and a single case of bilateral macular deposits in KMS, but this is the first report of a Coats type retinal telangiectasia. This case highlights the importance of thorough posterior segment examination in cases of KMS. PMID- 16352480 TI - Bilateral complete isolated cryptophthalmos: a case report. AB - Cryptophthalmos is a condition of congenital eyelid malformation most commonly accompanied by syndactyly, urogenital anomalies, and cognitive impairments as in Fraser syndrome. We report on a patient with characteristic features consistent with autosomal dominant bilateral complete isolated cryptophthalmos. This patient represents only the sixth documented case of bilateral complete isolated cryptophthalmos. Defining characteristics of this variety are discussed, including bilateral central dimpling over the globes, normal eyebrow growth, and the absence of cognitive impairment. We introduce phenotypic features that distinguish bilateral isolated cryptophthalmos from other forms and discuss its relatively favorable prognosis. PMID- 16352481 TI - The relative contribution of the X chromosome to ocular phenotypes. AB - The X chromosome is unique, both in terms of functional expression and evolutionary history. Population frequencies for a minority of conditions, such as mental retardation, are directly related to the X chromosome. To explore these ideas, we investigated the general role of the X chromosome in ocular genetics through bioinformatic analysis of the distribution of eye-related genes in the human genome. The proportion of eye-disease loci located on the X chromosome compared to those eye diseases with an autosomal locus was calculated. The resultant figure (3.47) is lower than that calculated for mental retardation (9.74). A comparison between the number of X chromosome genes expressed in the eye compared to the number of autosomal genes expressed in the eye also did not reveal significant differences. Of all genes expressed in the eye, 2.9% are thought to be located on the X chromosome, fewer than found for the larger autosomes (which range from 10.1% to 3.6%). The eye's functional genetic components appear to be dispersed throughout the human genome, possibly to ensure survival in the event of significant cytogenetic derangement. PMID- 16352483 TI - Atrial fibrillation: what can we do, what should we do, and what must we do? PMID- 16352484 TI - Gender differences in the treatment and secondary prevention of CHD at population level. AB - OBJECTIVES: Treatment and secondary prevention measures, received by persons with coronary heart disease (CHD), are insufficiently known at the moment. The aim of this study was to investigate the state of treatment and secondary prevention of CHD in a population-based sample and to analyze possible gender differences in different age groups. DESIGN: 300 men and 300 women with CHD were identified from a nationally representative health examination survey with 88% participation rate, carried out in Finland in 2000-2001. RESULTS: Revascularization had been performed on 34% (95% confidence interval 29, 40%) of men and 13% (8, 18%) of women. Moreover, 76% (71, 81%) of the men and 63% (57, 69%) of the women used antithrombotic medications. Two thirds of both men and women used beta-blockers and one third lipid-lowering medication. Smoking was more common among men, whereas obesity and high total cholesterol concentration were more common among women. CONCLUSIONS: Secondary prevention of CHD is far from optimal and there are gender differences in the care of CHD. PMID- 16352485 TI - Tissue Doppler echocardiographic identification of ischemic etiology in patients with dilated cardiomyopathy. AB - OBJECTIVE: To compare the left ventricular function in patients with ischemic and nonischemic cardiomyopathy using tissue Doppler echocardiography (TDE). METHODS: We studied 30 patients after myocardial infarction (MI group), 30 patients with dilated cardiomyopathy (DCM group) and 60 healthy volunteers in corresponding control groups. TDE velocities, time intervals of cardiac cycle were measured and dispersion index of TDE parameters was calculated. RESULTS: Early diastolic velocities were lower in MI group than in DCM group, with similar systolic and late diastolic velocities. The dispersion index of systolic velocities was significantly higher in MI than in DCM group and in controls (respectively 33.1+/ 6.0% vs 12.6+/-3.7% vs 15.9+/-5.6%; p < 0.001) and differentiated ischemic from idiopathic dilated cardiomyopathy. In MI group, preejection period was shorter and isovolumic relaxation and diastasis time were longer than in DCM group, with no differences in dispersion index of time intervals between the groups. CONCLUSIONS: TDE parameters: early diastolic velocity, preejection period, isovolumic relaxation time and the dispersion index of systolic velocities differentiate ischemic and nonischemic etiology of dilated cardiomyopathy. PMID- 16352486 TI - Epicardial and endocardial dispersion of ventricular repolarization. A study of monophasic action potential mapping in healthy pigs. AB - OBJECTIVES: To investigate the total dispersion of ventricular repolarization of the epi- and endocardium. DESIGN: Monophasic action potentials (MAP) were recorded from 211+/-54 (151-353) left and right ventricular epi- and endocardial sites during atrial pacing in 10 pigs using the CARTO system. The activation time (AT), MAP duration (MAPd) and end of repolarization time (EOR) were measured. RESULTS: The total dispersion of AT, EOR and MAPd, defined as the maximal differences of these parameters over both the epi- and endocardium, were 57+/-10, 84+/-20, and 75+/-21 ms respectively and were significantly larger than the respective epi- and endocardial dispersions (p<0.05). The epicardial dispersion of AT, EOR and MAPd of both the right and left ventricles were significantly larger than that of each ventricle alone (p<0.02). Sternotomy did not affect these dispersion parameters. CONCLUSION: Detailed mapping of epicardial repolarization in vivo using the MAP mapping technique is feasible. Both the epi- and endocardium of the two ventricles contribute significantly to the total dispersion of repolarization. PMID- 16352487 TI - Nicotine inhibits large conductance Ca(2+)-activated K(+) channels and the NO/ cGMP signaling pathway in cultured human endothelial cells. AB - OBJECTIVE: The effects of nicotine on endothelium-dependent vasorelaxation mediated by nitric oxide (NO) are controversial. Since endothelial NO synthesis has been shown to depend on the activity of large conductance Ca(2 + )-activated K(+) channels (BK(Ca)), the present study investigated whether nicotine alters BK(Ca) single channel activity induced by the K(+) channel opener NS1619, and to examine a possible interaction with the endothelial NO generation. DESIGN: The patch-clamp technique was used to examine the BK(Ca) activity. NO production was measured indirectly using a [(3)H]-cGMP-radioimmunoassay. All experiments were performed using cultured endothelial cells derived from human umbilical cord veins. RESULTS: The BK(Ca) opener NS1619 (10 micromol/l) significantly increased the BK(Ca) open-state probability (NPo) from 0.011+/-0.007 (control) to 0.052+/ 0.019. Co-perfusion with nicotine (1 micromol/l) significantly decreased NS1619 induced NPo (n = 14, p < 0.05). Intracellular cGMP levels were significantly increased, if cells were stimulated with NS1619 (+ 225%; n = 10, p < 0.05), which was blocked by Nicotine (1 micromol/l). CONCLUSIONS: The results of the present study demonstrate that BK(Ca) activation by NS1619 plays an important role in the regulation of the NO-/cGMP-signaling-pathway. Endothelial dysfunction caused by nicotine may be connected with a decrease in BK(Ca)-activity. PMID- 16352488 TI - Vascular endothelial growth factor in coronary sinus: evidence for its association with coronary collaterals. AB - OBJECTIVES: To explore whether local growth factors concentration, including vascular endothelial growth factor (VEGF) and transforming growth factor beta one (TGF-beta(1)), influence the formation of coronary collaterals. DESIGN: Thirty six patients scheduled for coronary angiography received a 6F Goodale-Lubin catheterization to collect blood from the coronary sinus (CS) and right atrium (RA). RESULTS: Patients with coronary collaterals had a higher number of diseased vessels (2.6+/-0.2 vs. 1.4+/-0.3, p = 0.005), higher percentage of severity of stenosis (93+/-2 vs. 48+/-8, P < 0.001) and higher VEGF concentrations in CS (38.9+/-3.9 pg/ml vs. 20.8+/-1.4 pg/ml, P < 0.001) and in RA (31.7+/-3.1 pg/ml vs. 22.0+/-2.3 pg/ml, p = 0.004). There was no significant relationship between coronary collateral formation and TGF-beta(1) concentration. By binary logistic regression analysis, VEGF concentrations in CS (p = 0.030) and stenosis severity (p = 0.042) are correlated positively with collateral formation. CONCLUSIONS: The association between local, endogenous secretion of VEGF and coronary collateral formation is compatible with a paracrine role for this growth factor in pathophysiologic collateral formation. PMID- 16352489 TI - Leukocyte depleting filter attenuates myocardial injury during elective coronary artery bypass surgery. AB - BACKGROUND: Ischemia-reperfusion injury secondary to leukocyte activation has been widely recognized as one of the most relevant mechanism leading to postoperative organ dysfunction occurring after a period of ischemia. The aim of the present study was to evaluate in a prospective, randomized study, the value of leukocyte depleting filter in patients undergoing elective coronary artery bypass surgery. METHODS: Twenty patients scheduled for elective on-pump coronary artery bypass surgery were randomized to undergo cardiopulmonary bypass either with a leukocyte depleting filter incorporated in the extracorporeal circulation arterial line or without a filter. RESULTS: The main finding of this study was the significantly lower postoperative concentrations of cardiac troponin I in the leukocyte filter group (Tests of between-subjects effects: p = 0.024). There were also slightly better cardiac indices in the leukocyte filter group. A larger amount of blood units was infused intra- and postoperatively in patients undergoing cardiopulmonary bypass with leukocyte filtration (median, 600 [IQR, 0 1200] vs. 0 [IQR, 0-600], p = 0.08). Two patients in the leukocyte filter group underwent reoperation for bleeding but none in the control group (p = 0.48). Intra-and postoperative platelet count was lower in the leukocyte filter group (Tests of between-subjects effects: p = 0.08). Despite a significant increased concentration of C-reactive protein on the first postoperative day in the control group (p = 0.029), repeated-measures analysis failed to show any significant increase during the study period (p = 0.33). CONCLUSIONS: The results of this study suggest a myocardial protective effect of leukocyte filter in the setting of elective coronary artery bypass surgery. PMID- 16352490 TI - Antithrombotic- and anticoagulation regimens in OPCAB surgery. A Nordic survey. AB - OBJECTIVES: The aim of the survey was to determine current practices for antiplatelet- and heparin therapy in OPCAB surgery in the Nordic countries. DESIGN: A postal survey questionnaire was sent to all 26 departments of Cardiothoracic Surgery in Norway, Sweden, Iceland, Finland, and Denmark. RESULTS: The overall response rate was 100%. The respondents performed between 130 and 1000 CABG procedures per year. Between 0.3% and 42.0% of these procedures were performed as OPCAB. Preoperative antiplatelet therapy was administered by 48%, while 96% routinely administer postoperative antiplatelet therapy. In the postoperative antiplatelet therapy there was a trend toward a combination of aspirin and low molecular weight heparin. Perioperative anticoagulation practices with heparin varied among departments as well as reversal with protamine. Eighty percent of the respondents register postoperative complications. CONCLUSION: In the Nordic countries there is no uniform practice regarding antiplatelet therapy pre- and postoperatively as well as heparin therapy perioperatively. Dose regimens are highly variable and standard protocols have not yet been developed. PMID- 16352491 TI - Delayed uncoupling is related to cardioprotection induced by kappa-agonist U 50,488H in rat heart. AB - OBJECTIVES: To determine whether the kappa-opioid receptor agonist U50,488H affects electrical uncoupling during prolonged ischemia and, if so, whether the changes are associated with its cardioprotective action. DESIGN: The isolated rat heart was perfused in a Langendorff apparatus. Formazan content, lactate dehydrogenase (LDH) and hemodynamic parameters were measured to confirm the cardioprotective effect of U50,488H. The effects of U50,488H on electrical coupling during prolonged ischemia were also measured. RESULTS: U50,488H concentration-dependently increased formazan content and reduced LDH release, and the ameliorating effect of 10(-5) mol/L U50,488H was abolished by 5 x 10(-6) mol/L nor-binaltorphimine (nor-BNI), a selective kappa-opioid receptor antagonist, or 10(-4) mol/L 5-hydroxydecanoate (5-HD), a selective mitochondrial ATP-sensitive K(+) (K(ATP)) channel blocker. The onset of electrical uncoupling during prolonged ischemia was delayed by U50,488H, and the delay was not only abolished, but also advanced by nor-BNI or 5-HD relative to the control group. CONCLUSIONS: These results demonstrate that delayed uncoupling during prolonged ischemia is associated with the cardioprotection of U50,488H, and these effects of U50,488H are mediated by mitochondrial K(ATP) channels. PMID- 16352492 TI - Glaucoma in retinoblastoma. AB - The association between retinoblastoma and secondary glaucoma is well known. The most common cause of secondary glaucoma in retinoblastoma is iris neovascularization (NVI) followed by pupillary block and tumor seeding of the anterior chamber. Although glaucoma is a secondary clinical issue in retinoblastoma (RB) management and care, awareness of its presence, revealed by a thorough ocular exam of the anterior segment, can guide the clinician in assessing the overall condition of the affected eye. PMID- 16352493 TI - Intraocular choristoma. AB - Choristomas have been reported in the orbit, cornea, conjunctiva, uvea, retina, and optic nerve. Many reports have described the choristoma mainly in the epibulbar area, but little is known of the choristoma that occurs intraocularly. We reviewed the literature and summarized the reports that described the choristoma inside the eyes. Iris, ciliary body, choroid, and optic nerve head are the commonly affected tissues. In the anterior uveal tissue, an ectopic lacrimal gland is common and appears as a pinkish, nodular, fleshy mass. Its clinical course largely depends on the enlargement of the accompanying cyst. Osseous choristoma is mainly found in the choroid in young female patients. Although it is regarded as a benign tumor, it grows progressively and can decrease visual function. Treatment consists of close observation and possibly surgical intervention. Fortunately, most of the intraocular choristomas do not need aggressive treatment. PMID- 16352494 TI - Genomic investigations of posterior uveal melanoma. AB - The specific genetic mechanisms responsible for the malignant behavior of uveal melanoma are not known. Unlike cutaneous melanoma, epidemiologic studies have not demonstrated a definitive germline form of uveal melanoma, though familial melanoma and racial predilections occur. Molecular cytogenetic characterization of uveal melanoma suggests that somatic deletions of chromosome 3 are associated with a worse prognosis. Microarray technology has been used to characterize uveal melanoma gene expression and may provide tests useful for determining prognosis. As an improved understanding of the cellular mechanisms used by uveal melanoma is gained, new opportunities to adapt or design therapeutic approaches may emerge. PMID- 16352495 TI - Treatment of conjunctival lymphomas. AB - Due to their indolent nature, conjunctival lymphomas have different treatment options compared to other adnexal lymphomas. Therapeutic options directed towards conjunctival lymphomas include radiotherapy (external beam radiation), brachytherapy, cryotherapy, intralesional interferon injections, systemic rituximab, and observation. Radiotherapy that has been demonstrated to be highly effective for both low and high-grade tumors, however, can be associated with a high percentage of short-term side effects and long-term complications. Brachytherapy can provide local tumor control, but can also cause many complications. Cryotherapy may be used for certain tumor locations, but has a high recurrence rate. Both interferon and rituximab show promising results for treating low-grade tumors while avoiding the complications associated with radiotherapy. Finally, reports of spontaneous regression of low-grade conjunctival lymphomas have led some investigators to advocate a course of careful observation following biopsy. PMID- 16352496 TI - Preoperative cranial ultrasound findings in infants with major congenital heart disease. AB - BACKGROUND: Advances in diagnostic testing and surgical techniques have resulted in reduced mortality in neonates with congenital heart disease (CHD) and a major concern for neurological morbidity in the presence of preoperative neurological injury. OBJECTIVES: To determine the incidence and nature of preoperative cerebral ultrasound abnormalities in neonates with major CHD and to examine the relationship between cerebral abnormalities and the type of CHD. METHODS: Retrospective study; inclusion criteria: (1) neonates with major CHD admitted to the NICU over a 3-y period, (2) gestational age >35 wk, (3) documented preoperative cranial ultrasound available; exclusion criteria: (1) small for gestational age, (2) other congenital anomalies and/or chromosomal abnormalities, (3) a 5-min Apgar score <7, (4) congenital infection. Cranial ultrasounds (CUS) were reviewed without knowledge of the cardiac defect. CHDs were categorized. RESULTS: Fifty of 108 neonates with CHD met the inclusion criteria. Twenty-one patients (42%) had abnormalities on CUS. Thirteen of these (26%) had widened ventricular and/or subarachnoid spaces, three (6%) lenticulostriate vasculopathy, one (2%) calcification in the basal nuclei, and four (8%) had acute ischaemic changes. Cerebral abnormalities occurred more frequently in patients with coarctation or hypoplastic left heart syndrome (HLHS) than transposition of the great arteries (TGA) (63% vs 14%; n.s.). CONCLUSION: There is a high incidence of preoperative cerebral ultrasound abnormalities in this group of neonates with major CHD. PMID- 16352497 TI - Developmental delay at 12 months in children born extremely preterm. AB - AIM: To evaluate the feasibility and validity of a structured telephone interview to assess the development of children born extremely preterm. METHODS: The parents of 88 children born with a gestational age below 28 wk admitted to the neonatal intensive care unit (NICU) at Rigshospitalet, Copenhagen, were interviewed by telephone when their child was 1 y of age, corrected for preterm birth. A fully structured questionnaire on psychomotor function was used (Revised Prescreening Developmental Questionnaire (R-PDQ)). The parents of 30 children born at term without complications were interviewed for comparison. The interview was conducted by NICU staff. To validate the R-PDQ, parents of 22 children in the preterm group and parents of 19 children in the reference group conducted an Ages and Stages Questionnaire (ASQ) when their children had reached the age of 3 3(1/2) y. RESULTS: The R-PDQ was easy to use by staff and well accepted by parents. The mean score in the preterm group was 14.9+/-3.9 vs 17.7+/-2.7 in the term group (p<0.001). Three children had developmental scores below-2 SD. The R PDQ score was associated with the ASQ score 2 y later. CONCLUSION: A structured questionnaire administrated by telephone is an alternative and usable tool for assessing neurodevelopmental deficit in children born extremely preterm. The mean developmental delay in the preterm group compared to the term group (about-1 SD) was close to expectations. PMID- 16352498 TI - Tourette syndrome and other tic disorders in a total population of children: clinical assessment and background. AB - AIM: To describe the symptoms, onset, heredity, pre-/perinatal events and socio economic status in Tourette syndrome (TS) and other tic disorders. METHODS: From a total population of 4479 children, 25 (0.6%) with TS, 58 (1.3%) with chronic motor/vocal tics (CMVT) and 214 (4.8%) with transient tics (TT) in the last year were found. A three-stage procedure was used: tic screening, telephone interview and clinical assessment. The TS group was compared with 25 children with TT and 25 controls without tics. RESULTS: The mean age of the first symptoms of TS was significantly lower than the onset of CMVT. All except one with TS had contact with medical services. The tics of children with TS were significantly more severe than the tics of others. Younger age of onset of TS indicated more severe tics. Parents and siblings of children with TS had an increased prevalence of tic disorders, obsessive-compulsive behaviour (OCD), attention-deficit/hyperactivity disorder (ADHD) and depression. Eighty per cent had a first-degree relative with a psychiatric disorder. A non-significant increase with regards to reduced optimality score in the pre-, peri- or neonatal periods was found in children with TS compared to controls. No differences were found concerning socio-economic status. CONCLUSION: Almost all children from a total population with TS have sought help from medical services. An increased prevalence of tics, OCD, depression or ADHD was found in the parents/siblings of children with TS, which draws attention to the importance of thorough investigation of family members. PMID- 16352499 TI - Self-determination: can we let the child determine the 'best' treatment? PMID- 16352500 TI - Treatment of developmental dyslexia: a review. AB - Remarkably few research articles on the treatment of developmental dyslexia were published during the last 25 years. Some treatment research arose from the temporal processing theory, some from the phonological deficit hypothesis and some more from the balance model of learning to read and dyslexia. Within the framework of that model, this article reviews the aetiology of dyslexia sub types, the neuropsychological rationale for treatment, the treatment techniques and the outcomes of treatment research. The possible mechanisms underlying the effects of treatment are discussed. PMID- 16352501 TI - An evaluation of the responsiveness of a comprehensive set of outcome measures for children and adolescents with traumatic brain injuries. AB - The relative responsiveness of nine outcome measure scales was evaluated with 33 children and adolescents (aged 4-18 years) who had sustained traumatic brain injuries. Scales were selected to evaluate outcomes from each of the World Health Organization (WHO) International Classification of Functioning, Disability and Health domains. The outcome measures were administered to all participants during their inpatient rehabilitation stay and again at a follow-up clinic visit. No single outcome measure captured the diversity of improvement in this sample. The measures agreed that improvement had occurred, but did not agree about which children were improving. This result suggests that the scales were measuring different skills and outcomes. Three of the measures used in combination, either the Child Health Questionnaire or the Functional Independence Measure for Children, the American Speech-Language-Hearing Association National Outcome Measures System (Birth to Kindergarten NOMS/School-aged Health Care) and the Gross Motor Function Measure, are sufficient to detect change in each of the children where change occurred. The Pediatric Evaluation of Disability Inventory and the MultiAttribute Health Status Classification were the least responsive of the nine measures used. PMID- 16352502 TI - Sagittal counter forces (SCF) in the treatment of idiopathic scoliosis: a preliminary report. AB - BACKGROUND: In patients with idiopathic scoliosis (IS), reduced thoracic kyphosis and reduced lumbar lordosis frequently occur in correlation with the lateral spinal curvature. Normalization of the sagittal profile and hyper-correction of the deviation in frontal and coronal plane are the main issues of the latest concept of bracing. The purpose of this study was to investigate the influence of of sagittal counter forces (SCF) on the scoliotic deformity. STUDY DESIGN: A case series of four patients with IS treated with two braces designed to improve the sagittal profile (Rigo-System-Cheneau-brace and with a sagittal counter force brace, SCF-brace). METHODS: The short-term effect (30 min) of both braces was evaluated using surface topography (Formetric surface topography system, Diers International, Wiesbaden). RESULTS: One patient (Cobb angle 92 degrees ) showed no short-term correction in the frontal and coronal planes; others (Cobb angles between 39 and 48 degrees ) exhibited valuable correction in frontal and coronal planes. There was no short-term correction in the sagittal plane for either brace. CONCLUSION: The application of sagittal counter forces (SCF) seems to have similar short-term effects as 3D correction and should be addressed more in future concepts of scoliosis bracing. PMID- 16352503 TI - Voices from the past. A developmental psychiatrist considers Still's 'Some abnormal psychical conditions in children'. PMID- 16352504 TI - Paediatric rehabilitation in Italy. AB - In Italy, paediatric rehabilitation is part of the activity of services for neurological and psychiatric diseases in childhood and adolescence. These services (UONPIA) have an integrated, multi-professional, child- and family oriented approach to paediatric disabling conditions. At present, inadequate territorial distribution of services, shortage of resources and need of epidemiological monitoring and specific designs of bio-medical research are the main problems. PMID- 16352505 TI - Torticollis and plagiocephaly in infancy: therapeutic strategies. AB - BACKGROUND: Asymmetry in infancy is a diagnosis with a large spectrum of features, expressing an abnormal shape of parts of the body or unequal postures and movements, which might be structural and/or functional, with localized or generalized expression. PURPOSE: The purpose of the present study is to highlight different therapeutic aspects of the most occurring asymmetries in infancy: congenital muscular torticollis, positional torticollis and plagiocephaly, based on best evidence in current literature. RESULTS: A flow chart is presented showing different pathways in therapeutic strategies, such as physical therapy, orthotic devices (helmet treatment and Dynamic Orthotic Cranioplasty) and surgery. CONCLUSION: It is concluded that there are different views towards management on torticollis and plagiocephaly. A systematic therapeutic management to evaluate these asymmetries is indicated. The presented therapeutic flow chart might serve as a basis in order to achieve uniformity in therapeutic thinking and performance. PMID- 16352506 TI - Costs and complications of hospitalizations for children with cerebral palsy. AB - BACKGROUND: Although many children with cerebral palsy (CP) develop secondary conditions requiring hospitalization, in-patient hospital utilization by this population has not been characterized. OBJECTIVE: To characterize hospitalizations in children with CP and to compare them with hospitalizations of those without CP using a large national data set. METHODS: Analysis of the Healthcare Utilization Project Kid Inpatient Database, a weighted survey of paediatric discharges from US hospitals in 1997. RESULTS: In 1997, 37,000 children with CP were hospitalized, generating charges approaching 600 million dollars. Children with CP demonstrated longer lengths of stay (6.3 vs 4.1 days, p < 0.001), higher total charges (16,024 vs 9952 dollars, p < 0.001), more diagnoses (5.6 vs 3.0, p < 0.001) and more procedures (1.7 vs 1.1, p < 0.001) per admission. Five major diagnostic categories accounted for 83.2% of the discharge diagnoses for children with CP (48.6% for those without CP, p < 0.001). Children with CP were more often transferred to other facilities (p < 0.001) or prescribed home health care (p < 0.001) upon discharge. CONCLUSIONS: Hospitalization of children with CP represents a major expenditure for health care systems. Studies to improve the management of conditions associated with CP could result in better outcomes for children and families and potentially decrease costs associated with hospitalization. PMID- 16352507 TI - An optic micro-switch for an eyelid response to foster environmental control in children with minimal motor behaviour. AB - This study assessed whether two children (11.9- and 9.7-years-old) with profound multiple disabilities and minimal motor behaviour could learn to control environmental stimulation using an eyelid response with a newly developed micro switch. The response consisted of raising the eyelid markedly (i.e. by looking upward or opening the eyes widely). The micro-switch developed for this target response consisted of an electronically regulated optic sensor mounted on an eyeglasses' frame. Data showed that the children learned the target eyelid response to activate the micro-switch and to increase their level of environmental stimulation. Responding was largely maintained at a 2-month post intervention check. These results indicate that continued work in this area has positive implications for the rehabilitation of children with most serious disabilities. PMID- 16352508 TI - Thermoregulation during exercise in severely burned children. AB - INTRODUCTION: Severe burns result in skeletal muscle catabolism and weakness, which is worsened by prolonged physical inactivity. Exercise would be an ideal tool in the rehabilitation of burned children. However, it has been postulated that burned children may have an excessive rise in body temperature during exercise compared to non-burned children, partly due to the reduced area available for heat dissipation, thereby questioning the safety of exercise in burned children. METHODS: Children (n = 15) with >40% total body surface area (TBSA) burns and non-burned children (n = 13) successfully completed this study. All subjects completed 20 minutes of treadmill exercise at approximately 75% of their peak aerobic power. Tympanic temperature (Ttym), burned and unburned skin temperature were recorded pre-exercise, every 2 minutes during exercise and during recovery. RESULTS: Within burned children, significant differences between the temperature of unburned skin and burned skin, during later stages of sub maximal exercise (minutes 12-20) were present. However, there were no significant differences between burned and non-burned children in Ttym or unburned skin temperature indicating that severely burned children do not demonstrate an impaired thermoregulatory response to 20 minutes of sub-maximal exercise at room temperatures. CONCLUSION: It is concluded that exercise at moderate intensities conducted at room temperature is safe in burned children with <75% TBSA burns. PMID- 16352509 TI - Outcome of intensive oral motor and behavioural interventions for feeding difficulties in three children with Goldenhar Syndrome. AB - OBJECTIVE: The facial anomalies and surgeries associated with Goldenhar Syndrome often lead to feeding problems. The purpose of this study was to demonstrate the outcome of Goldenhar Syndrome children admitted to the day patient paediatric feeding programme. SUBJECTS: Three children with Goldenhar Syndrome and feeding difficulties participated, mean age 3 years. Average length of stay was 8 weeks. Upon admission all were dependent on tube feedings. METHODS: The children received four therapeutic sessions daily. The sessions included oral motor interventions, behavioural techniques and a structured meal. Data regarding children's feeding were collected at admission and discharge. RESULTS: The patients had 52% acceptance of food/drink at admission and 88.7% at discharge. Expels were 30% at admission and 14% at discharge. Mouth clean was 9.3% at admission and 97% at discharge. Total inappropriate feeding behaviours averaged 83% at admission and 27.3% at discharge. Children averaged 3.4 g per meal at admission and 105.4 g at discharge. Two children were completely weaned from tube feedings by discharge. CONCLUSIONS: Results support the hypothesis that combining oral motor and behavioural interventions offer effective treatment for children with Goldenhar Syndrome. Many children with Goldenhar Syndrome are reliant on tube feedings and this regimen allows them to eat orally, thereby improving their nutrition and quality of life. PMID- 16352510 TI - Paediatric rehabilitation in Israel: unique problems, exceptional solutions. AB - Mind the gap: Israel straddles the gap between Western medicine and Middle Eastern cultural and political challenges. Maintaining state-of-the-art paediatric rehabilitation in an era of economic and political constrictions is described and analysed in this review. PMID- 16352512 TI - A call for violence prevention in 2006. PMID- 16352513 TI - No man's land: gender bias and social constructivism in the diagnosis of borderline personality disorder. AB - The literature on borderline personality disorder (BPD), including its epidemiology, biology, phenomenology, causes, correlates, consequences, costs, treatments, and outcomes is vast. Thousands of articles and books have been published. Because the true prevalence of BPD by sex (gender) in the general population is still unknown, the important question of why women, rather than men, are more frequently diagnosed with BPD remains largely unanswered-despite current evidence for the origin of personality disorder in genetics and neurobiology, and despite recent suggestions that biased sampling is the most likely explanation for gender bias in the diagnosis of BPD. This paper reviews selected literature on (a) the epidemiology of BPD, (b) gender bias in the diagnosis of BPD, and (c) the social construction of diagnosis, particularly the diagnostic entity labeled "Borderline Personality Disorder." It attempts a synthesis of diverse, multidisciplinary literature to address the question of why women outnumber men by a ratio of 3:1 in the diagnosis of BPD. It rests on assumptions that (a) to varying degrees sociocultural factors inevitably play a role in the expression of disease conditions, and that (b) personality disorders, including BPD, have cultural histories. It also rests on the belief, for which there is considerable scholarly support, that the phenomenon called BPD has multiple, complex, interactive, biological, psychological, and constructed sociocultural determinants. Nurses must understand the phenomenon at this level of complexity to provide appropriate care. PMID- 16352514 TI - Living among strangers: the needs and functioning of persons with schizophrenia residing in an assisted living facility. AB - OBJECTIVES: The purposes of this research were to describe the quality of care for severe and persistent mentally ill persons with schizophrenia who reside in an assisted living facility and to explore the characteristics related to the identification of need and the level of functioning of these individuals. The specific objectives of the study were to (a) describe characteristics of the residents, (b) identify relationships among characteristics of residents and the level of functioning, and (c) analyze relationships among these characteristics and resident needs. DESIGN: Correlational, quasi-experimental design. POPULATION/ SAMPLE/SETTING: Face-to-face semistructured interviews were conducted with 58 ALF residents with schizophrenia and eight direct caregivers from eight assisted living facilities in Florida, resulting in representation of 20% of the public assisted living facilities in one county. METHOD: Face to face, semi-structured interviews utilizing multiple scales and instruments. FINDINGS: Findings indicated that the profile of a typical resident living in a public ALF consisted of a 43-year-old, never married Caucasian male who dropped out of high school and had suffered from a psychiatric illness for over 21 years. The most frequent reported current mental health treatment modality of residents was attendance at a community mental health center. This average resident had five inpatient psychiatric hospitalizations, had not been an inpatient at a state psychiatric facility, and was prescribed an average of four different psychiatric medications. Spearman correlational coefficients and stepwise multiple regression analyses revealed that significant relationships existed between level of functioning and the following characteristics: positive symptoms (p = 0.0003), negative symptoms (p = 0.003), medication knowledge (p = 0.0001), and GAF scores, current (p = 0.01) and past (p = 0.009). No significant relationships were indicated among resident characteristics and resident needs. CONCLUSIONS: This descriptive, correlational study provides the foundation for further research on the quality of care for individuals with severe and persistent mental illnesses who reside in assisted living facilities. Understanding the interactions among symptoms of schizophrenia, level of functioning, and unmet needs may provide a future focus for mental health treatment planning and intervention. IMPLICATIONS: National and international agencies develop and implement assisted living facility standards that impact the lives of residents with moderate levels of functioning and multiple unmet needs, having implications for both practice and mental health policy. PMID- 16352515 TI - The healing circle: resiliency in nurses. AB - The concept of resiliency has been explored extensively in the fields of developmental psychology as an adaptive life process. Increasingly nurses have begun to study resiliency in a wide variety of settings and client populations. This article explores the concept of resiliency in nurses. Resiliency was described through the use of personal exemplar, tracing the author's odyssey of nursing homeless men in an emergency shelter. The author proposes that the traits of resiliency in nurses are widespread and largely unrecognized. There is a pressing need to cultivate and foster personal growth in nurses because we cannot give our patients what we do not possess ourselves. PMID- 16352516 TI - Raising the awareness of primary care providers about postpartum depression. AB - About 13% of women experience depression in the first year after childbirth. Postpartum depression has deleterious effects on the woman's relationships, her functional status, and her ability to care for her infant. Primary care providers have the most contact with postpartum women, but may be unable or unwilling to screen, treat, and/or refer the women. Thus, many women with postpartum depression are not receiving mental health services. The purpose of this article is to describe methods to raise the awareness of primary care providers about postpartum depression, thereby eliminating a major barrier to mental health treatments of postpartum women. PMID- 16352517 TI - A study of psychiatric nurses' beliefs and attitudes about work safety and assaults in Turkey. AB - In Turkey, the cultural structure effects the gender roles in the society, and women are subject to violence in an intense manner both inside the family and in their workplaces. In nursing, which is still defined as a woman's job in our country, it is possible to encounter many aggressive and threatening behaviours. Knowing no geographical borders, aggression leads to dissatisfaction and alienation with the profession for the nurses working at the psychiatric institutions in Turkey, thus aggression is a significant health risk in physical, psychological and social dimensions. The research was carried out with 162 randomly selected nurses working in the psychiatric institutions in Istanbul using the Attitudes Toward Patient Physical Assault Questionnaire. It has been determined that nurses' rate of exposure to verbal/physical assault by the patients and their relatives is high and that they experience an intense uncertainty and conflict, especially in legal issues, and that the duration of working in the psychiatry clinics effects attitudes toward the assaults. It is concluded that the nurses working at the psychiatry clinics in Turkey are under risk in regards to safety and they need protection and support, both in emotional and in legal terms. PMID- 16352518 TI - Exploring the role of the mental health nurse in community mental health care for the aged. AB - There is currently considerable discussion about the impact of the aging population on the demand for health care services, however there is considerably less attention paid to the impact of mental health issues on the needs of the aged population. Nurses comprise the largest professional group within the mental health workforce in Australia. The availability of a high quality mental health nursing workforce will therefore be crucial to meeting the health needs of aging clients in the future, accompanied by an increased pressure to increase the proportion of care delivered in the community. There is however, a paucity of literature on the role and contribution of community mental health nurses specialising in the aged care field. The aim of this paper is to present the findings of a project designed to examine the role of mental health nursing within aged persons' community mental health teams in Victoria, Australia, with particular emphasis on the biopsychosocial interventions used. Fifteen participants from three community mental health services in Victoria participated in a focus group interview to share their insights and experiences. Data analysis revealed two main themes, the role of the nurse, and the specific functions of the nurse. This data is presented as a beginning contribution to the paucity of literature currently available in this important area. PMID- 16352522 TI - Dynamic spectrum quality assessment and iterative computational analysis of shotgun proteomic data: toward more efficient identification of post translational modifications, sequence polymorphisms, and novel peptides. AB - In mass spectrometry-based proteomics, frequently hundreds of thousands of MS/MS spectra are collected in a single experiment. Of these, a relatively small fraction is confidently assigned to peptide sequences, whereas the majority of the spectra are not further analyzed. Spectra are not assigned to peptides for diverse reasons. These include deficiencies of the scoring schemes implemented in the database search tools, sequence variations (e.g. single nucleotide polymorphisms) or omissions in the database searched, post-translational or chemical modifications of the peptide analyzed, or the observation of sequences that are not anticipated from the genomic sequence (e.g. splice forms, somatic rearrangement, and processed proteins). To increase the amount of information that can be extracted from proteomic MS/MS datasets we developed a robust method that detects high quality spectra within the fraction of spectra unassigned by conventional sequence database searching and computes a quality score for each spectrum. We also demonstrate that iterative search strategies applied to such detected unassigned high quality spectra significantly increase the number of spectra that can be assigned from datasets and that biologically interesting new insights can be gained from existing data. PMID- 16352523 TI - Characterization of an RNA granule from developing brain. AB - In brain, mRNAs are transported from the cell body to the processes, allowing for local protein translation at sites distant from the nucleus. Using subcellular fractionation, we isolated a fraction from rat embryonic day 18 brains enriched for structures that resemble amorphous collections of ribosomes. This fraction was enriched for the mRNA encoding beta-actin, an mRNA that is transported in dendrites and axons of developing neurons. Abundant protein components of this fraction, determined by tandem mass spectrometry, include ribosomal proteins, RNA binding proteins, microtubule-associated proteins (including the motor protein dynein), and several proteins described only as potential open reading frames. The conjunction of RNA-binding proteins, transported mRNA, ribosomal machinery, and transporting motor proteins defines these structures as RNA granules. Expression of a subset of the identified proteins in cultured hippocampal neurons confirmed that proteins identified in the proteomics were present in neurites associated with ribosomes and mRNAs. Moreover many of the expressed proteins co localized together. Time lapse video microscopy indicated that complexes containing one of these proteins, the DEAD box 3 helicase, migrated in dendrites of hippocampal neurons at the same speed as that reported for RNA granules. Although the speed of the granules was unchanged by activity or the neurotrophin brain-derived neurotrophic factor, brain-derived neurotrophic factor, but not activity, increased the proportion of moving granules. These studies define the isolation and composition of RNA granules expressed in developing brain. PMID- 16352524 TI - Docosahexaenoic acid facilitates cell maturation and beta-adrenergic transmission in astrocytes. AB - The effects of docosahexaenoic acid (DHA; 22:6 n-3), a major omega-3 PUFA in the mammalian brain, on the structure and function of astrocytes were studied using primary cultures from rat cerebra. Gas-liquid chromatography of methyl esters of FAs isolated from cultures exposed to individual FAs, namely, stearic acid, linoleic acid, arachidonic acid, and DHA, showed alterations in the lipid profiles of the membranes, with a preferential incorporation of the FA to which the cells were exposed. Immunofluorescence studies demonstrated that unlike treatment with other FAs, after which the astrocytes remained as immature radial forms, DHA-treated astrocytes showed distinct differentiation, having morphology comparable to those grown in normal serum-containing medium. Receptor binding studies to determine the concentration of various neurotransmitter receptors showed that DHA selectively increased the number of beta-adrenergic receptors (beta-ARs) compared with FA-untreated controls, suggesting a greater role of DHA on beta-AR expression in membranes. This was also reflected by an increase in downstream events of the beta-AR pathways, such as the induction of protein kinase A and glycogen turnover by isoproterenol (ISP), a beta-AR agonist in DHA treated cells. Moreover, ISP completely transformed DHA-treated cells into mature astrocytes bearing long processes, as in cells grown under normal conditions. Together, our observations suggest that DHA plays a unique role in facilitating some of the vital functions of astrocytes in the developing brain. PMID- 16352525 TI - Complete predicted three-dimensional structure of the facilitator transmembrane protein and hepatitis C virus receptor CD81: conserved and variable structural domains in the tetraspanin superfamily. AB - Tetraspanins are a superfamily of transmembrane proteins implicated in cellular development, motility, and activation through their interactions with a large range of proteins and with specific membrane microdomains. The complete three dimensional structure of the tetraspanin CD81 has been predicted by molecular modeling and from the crystallographic structure of the EC2 large extracellular domain. Periodicity of sequence conservation, homology modeling, secondary structure prediction, and protein docking were used. The transmembrane domain appears organized as a four-stranded left-handed coiled coil directly connecting to two helices of the EC2. A smaller extracellular loop EC1 contains a small largely hydrophobic beta-strand that packs in a conserved hydrophobic groove of the EC2. The palmitoylable intracellular N-terminal segment forms an amphipathic membrane-parallel helix. Structural variability occurs mainly in an hypervariable subdomain of the EC2 and in intracellular regions. Therefore, the variable interaction selectivity of tetraspanins originates both from sequence variability within structurally conserved domains and from the occurrence of small structurally variable domains. In CD81 and other tetraspanins, the numerous membrane-exposed aromatic residues are asymmetrically clustered and protrude on one side of the transmembrane domain. This may represent a functional specialization of these two sides for interactions with cholesterol, proteins, or membrane microdomains. PMID- 16352527 TI - Examining the theory of error catastrophe. PMID- 16352528 TI - Regulatable gutless adenovirus vectors sustain inducible transgene expression in the brain in the presence of an immune response against adenoviruses. AB - In view of recent serious adverse events and advances in gene therapy technologies, the use of regulatable expression systems is becoming recognized as indispensable adjuncts to successful clinical gene therapy. In the present work we optimized high-capacity adenoviral (HC-Ad) vectors encoding the novel tetracycline-dependent (TetOn)-regulatory elements for efficient and regulatable gene expression in the rat brain in vivo. We constructed two HC-Ad vectors encoding beta-galactosidase (beta-gal) driven by a TetOn system containing the rtTAS(s)M2 transactivator and the tTS(Kid) repressor under the control of the murine cytomegalovirus (mCMV) (HC-Ad-mTetON-beta-Gal) or the human CMV (hCMV) promoter (HC-Ad-hTetON-beta-Gal). Expression was tightly regulatable by doxycycline (Dox), reaching maximum expression in vivo at 6 days and returning to basal levels at 10 days following the addition or removal of Dox, respectively. Both vectors achieved higher transgene expression levels compared to the expression from vectors encoding the constitutive mCMV or hCMV promoter. HC-Ad mTetON-beta-Gal yielded the highest transgene expression levels and expressed in both neurons and astrocytes. Antivector immune responses continue to limit the clinical use of vectors. We thus tested the inducibility and longevity of HC-Ad mediated transgene expression in the brain of rats immunized against adenovirus by prior intradermal injections of RAds. Regulated transgene expression from HC Ad-mTetON-beta-Gal remained active even in the presence of a significant systemic immune response. Therefore, these vectors display two coveted characteristics of clinically useful vectors, namely their regulation and effectiveness even in the presence of prior immunization against adenovirus. PMID- 16352529 TI - Herpes simplex virus DNA synthesis is not a decisive regulatory event in the initiation of lytic viral protein expression in neurons in vivo during primary infection or reactivation from latency. AB - The herpes simplex virus genome can enter a repressed transcriptional state (latency) in sensory neurons of the host nervous system. Although reduced permissiveness of the neuronal environment is widely accepted as a causal factor, the molecular pathway(s) directing and maintaining the viral genome in the latent state remains undefined. Over the past decade, the field has been strongly influenced by the observations of Kosz-Vnenchak et al., which have been interpreted to indicate that, in sensory neurons in vivo, a critical level of viral DNA synthesis within the neuron is required for sufficient viral immediate early (IE) and early (E) gene expression (M. Kosz-Vnenchak, J. Jacobson, D. M. Coen, and D. M. Knipe, J. Virol. 67:5383-5393, 1993). The levels of IE and E genes are, in turn, thought to regulate the decision to enter the lytic cycle or latency. We have reexamined this issue using new strategies for in situ detection and quantification of viral gene expression in whole tissues. Our results using thymidine kinase-null and rescued mutants as well as wild-type strains in conjunction with viral DNA synthesis blockers demonstrate that (i) despite inhibition of viral DNA replication, many neurons express lytic viral proteins, including IE proteins, during acute infection in the ganglion; (ii) at early times postinoculation, the number of neurons expressing viral proteins in the ganglion is not reduced by inhibition of viral DNA replication; and (iii) following a reactivation stimulus, the numbers of neurons and apparent levels of lytic viral proteins, including IE proteins, are not reduced by inhibition of viral DNA replication. We conclude that viral DNA replication in the neuron per se does not regulate IE gene expression or entry into the lytic cycle. PMID- 16352530 TI - Entry of vaccinia virus and cell-cell fusion require a highly conserved cysteine rich membrane protein encoded by the A16L gene. AB - The vaccinia virus A16L open reading frame encodes a 378-amino-acid protein with a predicted C-terminal transmembrane domain and 20 invariant cysteine residues that is conserved in all sequenced members of the poxvirus family. The A16 protein was expressed late in infection and incorporated into intracellular virus particles with the N-terminal segment of the protein exposed on the surface. The cysteine residues were disulfide bonded via the poxvirus cytoplasmic redox system. Unsuccessful attempts to isolate a mutant virus with the A16L gene deleted suggested that the protein is essential for replication. To study the role of the A16 protein, we made a recombinant vaccinia virus that has the Escherichia coli lac operator system regulating transcription of the A16L gene. In the absence of inducer, A16 synthesis was repressed and plaque size and virus yield were greatly reduced. Nevertheless, virus morphogenesis occurred and normal looking intracellular and extracellular virus particles formed. Purified virions made in the presence and absence of inducer were indistinguishable, though the latter had 60- to 100-fold-lower specific infectivity. A16-deficient virions bound to cells, but their cores did not penetrate into the cytoplasm. Furthermore, A16-deficient virions were unable to induce low-pH-triggered syncytium formation. The phenotype of the inducible A16L mutant was similar to those of mutants in which synthesis of the A21, A28, H2, or L5 membrane protein was repressed, indicating that at least five conserved viral proteins are required for entry of poxviruses into cells as well as for cell-cell fusion. PMID- 16352531 TI - Anti-CD45RO suppresses human immunodeficiency virus type 1 replication in microglia: role of Hck tyrosine kinase and implications for AIDS dementia. AB - Macrophages and microglia are productively infected by HIV-1 and play a pivotal role in the pathogenesis of AIDS dementia. Although macrophages and microglia express CD45, a transmembrane protein tyrosine phosphatase, whether modulation of its activity affects human immunodeficiency virus type 1 (HIV-1) replication is unknown. Here, we report that of the five human CD45 isoforms, microglia express CD45RB and CD45RO (RB > RO) and treatment of microglia with a CD45 agonist antibody alphaCD45RO (UCHL-1) inhibits HIV-1 replication. alphaCD45RO prevented HIV-1 negative factor (Nef)-induced autophosphorylation of hematopoietic cell kinase (Hck), a myeloid lineage-specific Src kinase. Recombinant CD45 protein also inhibited HIV-1-induced Hck phosphorylation in microglia. Antennapedia mediated delivery of Hck Src homology domain 3 (SH3), a domain that binds to the Nef PxxP motif with high affinity, reduced HIV-1-induced Hck phosphorylation and HIV-1 production in microglia. HIV-1-induced LTR transactivation was observed in U38 cells stably overexpressing wild-type Hck but not kinase-inactive Hck. In microglia, alphaCD45RO reduced activation of transcription factors (NF-kappaB and CCAAT enhancer binding protein) necessary for LTR transactivation in macrophages. These results establish that in myeloid lineage cells, Nef interacts with the Hck SH3 domain, resulting in autophosphorylation of Hck and an increase in HIV-1 transcription. alphaCD45RO-mediated inhibition of HIV-1 replication in microglia identifies the CD45 protein tyrosine phosphatase as a potential therapeutic target for HIV-1 infection/AIDS dementia. PMID- 16352532 TI - Functional domains of murine cytomegalovirus nuclear egress protein M53/p38. AB - Two conserved herpes simplex virus 1 proteins, UL31 and UL34, form a complex at the inner nuclear membrane which governs primary envelopment and nuclear egress of the herpesvirus nucleocapsids. In mouse cytomegalovirus, a member of the betaherpesvirus subfamily, the homologous proteins M53/p38 and M50/p35 form the nuclear egress complex (NEC). Since the interaction of these proteins is essential for functionality, the definition of the mutual binding sites is a prerequisite for further analysis. Using a comprehensive random mutagenesis procedure, we have mapped the M53/p38 binding site of M50/p35 (A. Bubeck, M. Wagner, Z. Ruzsics, M. Lotzerich, M. Iglesias, I. R. Singh, and U. H. Koszinowski, J. Virol. 78:8026-8035). Here we describe a corresponding analysis for the UL31 homolog M53/p38. A total of 72 individual mutants were reinserted into the genome to test the complementation of the lethal M53 null phenotype. The mutants were also studied for colocalization and for coprecipitation with M50/p35. The analysis revealed that the nonconserved N-terminal one-third of M53/p38 provides the nuclear localization signal as an essential function. The collective results for many mutants localized the binding site for M50/p35 to amino acids (aa) 112 to 137. No single aa exchange for alanine could destroy NEC formation, but virus attenuation revealed a major role for aa K128, Y129, and L130. The lethal phenotype of several insertion and stop mutants indicated the functional importance of the C terminus of the protein. PMID- 16352533 TI - Betanodavirus B2 is an RNA interference antagonist that facilitates intracellular viral RNA accumulation. AB - Betanodaviruses are small positive-sense bipartite RNA viruses that infect a wide variety of fish species and are notorious for causing lethal outbreaks in juvenile fish hatcheries worldwide. The function of a small nonstructural protein, B2, encoded by the subgenomic RNA3 of betanodaviruses, has remained obscure. Greasy grouper nervous necrosis virus, a betanodavirus model, was used to develop a facile DNA-based reverse genetics system that recapitulated the virus infection cycle, and we used this system to show that B2 is a small nonstructural protein that is essential for high level accumulation of viral RNA1 after RNA transfection of fish, mammalian, and avian cells. The defect in RNA1 accumulation in a B2 mutant was partially complemented by supplying B2 RNA in trans. Confocal analysis of the cellular distribution of B2 indicated that B2 is able to enter the nucleus and accumulates there during the late stages of GGNNV infection. Using human HeLa cells as a cellular RNA interference model, we found that B2 could efficiently antagonize RNA interference, which is a property shared by the distantly related alphanodavirus B2 proteins. This function provides appears to provide an explanation, at least in part, for why B2 mutant RNA1 is severely impaired in its intracellular accumulation. PMID- 16352534 TI - The equine herpesvirus 1 UL20 product interacts with glycoprotein K and promotes egress of mature particles. AB - The aim of the present study was to identify and functionally characterize the equine herpesvirus 1 (EHV-1) UL20 protein (UL20p). Using a specific antiserum, UL20p was shown to be associated with membranes of infected cells, as well as with envelopes of purified virions. By Western blot analysis, UL20p was detected in two main forms exhibiting M(r)s of 25,000 and 75,000. Both moieties did not enter the separating gel after heating of protein samples to 99 degrees C. The slower-migrating form of UL20p contains N-linked carbohydrates, and its presence is dependent of that of other viral proteins. Infection of cells that either constitutively express UL20p or a gK-green fluorescent protein (GFP) fusion protein with various EHV-1 deletion mutants revealed a relatively stable hetero oligomer containing gK and UL20p with an apparent M(r) of 75,000. As demonstrated by confocal microscopy, UL20p distribution in Rk13 cells changed from a diffuse granular or netlike appearance to a pattern confined to the Golgi network when gK was coexpressed. Analysis of a UL20 deletion mutant of EHV-1 strain RacL11 indicated an involvement of UL20p in cell-to-cell spread, as well as in very late events in virus egress. Based on these and electron microscopic studies we suggest that the EHV-1 UL20 protein might be necessary to avoid fusion of mature virions with membranes of their transport vesicles. PMID- 16352535 TI - Association of herpesvirus saimiri tip with lipid raft is essential for downregulation of T-cell receptor and CD4 coreceptor. AB - Lipid rafts are membrane microdomains that are proposed to function as platforms for both receptor signaling and trafficking. Our previous studies have demonstrated that Tip of herpesvirus saimiri (HVS), which is a T-lymphotropic tumor virus, is constitutively targeted to lipid rafts and interacts with cellular Lck tyrosine kinase and p80 WD repeat-containing endosomal protein. Through the interactions with Lck and p80, HVS Tip modulates diverse T-cell functions, which leads to the downregulation of T-cell receptor (TCR) and CD4 coreceptor surface expression, the inhibition of TCR signal transduction, and the activation of STAT3 transcription factor. In this study, we investigated the functional significance of Tip association with lipid rafts. We found that Tip expression remarkably increased lipid raft fractions in human T cells by enhancing the recruitment of lipid raft-resident proteins. Genetic analysis showed that the carboxyl-terminal transmembrane, but not p80 and Lck interaction, of Tip was required for the lipid raft localization and that lipid raft localization of Tip was necessary for the efficient downregulation of TCR and CD4 surface expression. Correlated with this, treatment with Filipin III, a lipid raft-disrupting agent, effectively reversed the downregulation of CD3 and CD4 surface expression induced by Tip. On the other hand, Tip mutants that were no longer present in lipid rafts were still capable of inhibiting TCR signaling and activating STAT3 transcription factor activity as efficiently as wild-type (wt) Tip. These results indicate that the association of Tip with lipid rafts is essential for the downregulation of TCR and CD4 surface expression but not for the inhibition of TCR signal transduction and the activation of STAT3 transcription factor. These results also suggest that the signaling and targeting activities of HVS Tip rely on functionally and genetically separable mechanisms, which may independently modulate T-cell function for viral persistence or pathogenesis. PMID- 16352536 TI - CD8+ T cells require perforin to clear West Nile virus from infected neurons. AB - Injury to neurons after West Nile virus (WNV) infection is believed to occur because of viral and host immune-mediated effects. Previously, we demonstrated that CD8+ T cells are required for the resolution of WNV infection in the central nervous system (CNS). CD8+ T cells can control infection by producing antiviral cytokines (e.g., gamma interferon or tumor necrosis factor alpha) or by triggering death of infected cells through perforin- or Fas ligand-dependent pathways. Here, we directly evaluated the role of perforin in controlling infection of a lineage I New York isolate of WNV in mice. A genetic deficiency of perforin molecules resulted in higher viral burden in the CNS and increased mortality after WNV infection. In the few perforin-deficient mice that survived initial challenge, viral persistence was observed in the CNS for several weeks. CD8+ T cells required perforin to control WNV infection as adoptive transfer of WNV-primed wild-type but not perforin-deficient CD8+ T cells greatly reduced infection in the brain and spinal cord and enhanced survival of CD8-deficient mice. Analogous results were obtained when wild-type or perforin-deficient CD8+ T cells were added to congenic primary cortical neuron cultures. Taken together, our data suggest that despite the risk of immunopathogenesis, CD8+ T cells use a perforin-dependent mechanism to clear WNV from infected neurons. PMID- 16352537 TI - "UnPAKing" human immunodeficiency virus (HIV) replication: using small interfering RNA screening to identify novel cofactors and elucidate the role of group I PAKs in HIV infection. AB - In order to identify novel proviral host factors involved in human immunodeficiency virus (HIV) infection, we performed a screen of a small interfering RNA (siRNA) library targeting 5,000 genes with the highest potential for being targets for therapeutics. Many siRNAs in the library against known host factors, such as TSG101, furin, and CXCR4, were identified as inhibitors by the screen and thus served as internal validation. In addition, many novel factors whose knockdown inhibited infection were identified, including Pak3, a member of the serine/threonine group I PAK kinases. The HIV accessory factor Nef has been shown to associate with a PAK kinase, leading to enhanced viral production; however, the exact identity of the kinase has remained controversial. Prompted by the Pak3 screen hit, we further investigated the involvement of group I PAK kinases in HIV using siRNA. Contrary to the current literature, Pak1 depletion strongly inhibited HIV infection in multiple cell systems and decreased levels of integrated provirus, while Pak2 depletion showed no effect. Overexpression of a constitutively active Pak1 mutant also enhanced HIV infection, further supporting its role as the dominant PAK involved. PMID- 16352538 TI - Soluble V domain of Nectin-1/HveC enables entry of herpes simplex virus type 1 (HSV-1) into HSV-resistant cells by binding to viral glycoprotein D. AB - Interaction of herpes simplex virus (HSV) glycoprotein D (gD) with specific cellular receptors is essential for HSV infection of susceptible cells. Virus mutants that lack gD can bind to the cell surface (attachment) but do not enter, implying that interaction of gD with its receptor(s) initiates the postattachment (entry) phase of HSV infection. In this report, we have studied HSV entry in the presence of the gD-binding variable (V) domain of the common gD receptor nectin 1/HveC to determine whether cell association of the gD receptor is required for HSV infection. In the presence of increasing amounts of the soluble nectin-1 V domain (sNec1(123)), increasing viral entry into HSV-resistant CHO-K1 cells was observed. At a multiplicity of 3 in the presence of optimal amounts of sNec1(123), approximately 90% of the cells were infected. The soluble V domain of nectin-2, a strain-specific HSV entry receptor, promoted entry of the HSV type 1 (HSV-1) Rid-1 mutant strain, but not of wild-type HSV-1. Preincubation and immunofluorescence studies indicated that free or gD-bound sNec1(123) did not associate with the cell surface. sNec1(123)-mediated entry was highly impaired by interference with the cell-binding activities of viral glycoproteins B and C. While gD has at least two functions, virus attachment to the cell and initiation of the virus entry process, our results demonstrate that the attachment function of gD is dispensable for entry provided that other means of attachment are available, such as gB and gC binding to cell surface glycosaminoglycans. PMID- 16352539 TI - A novel, highly selective inhibitor of pestivirus replication that targets the viral RNA-dependent RNA polymerase. AB - We report on the highly potent and selective antipestivirus activity of 5-[(4 bromophenyl)methyl]-2-phenyl-5H-imidazo[4,5-c]pyridine (BPIP). The 50% effective concentration (EC50) for inhibition of bovine viral diarrhea virus (BVDV)-induced cytopathic effect formation was 0.04 +/- 0.01 microM. Comparable reduction of viral RNA synthesis (EC50 = 0.12 +/- 0.02 microM) and production of infectious virus (EC50= 0.074 +/- 0.003 microM) were observed. The selectivity index (ratio of 50% cytostatic concentration/EC50) of BPIP was approximately 2,000. BPIP was inactive against the hepatitis C virus subgenomic replicon and yellow fever virus but demonstrated weak activity against GB virus. Drug-resistant mutants were at least 300-fold less susceptible to BPIP than wild-type virus; showed cross resistance to N-propyl-N-[2-(2H-1,2,4-triazino[5,6-b]indol-3-ylthio)ethyl]-1 propanamine (VP32947), and carried the F224S mutation in the viral RNA-dependent RNA polymerase (RdRp). When the F224S mutation was introduced into an infectious clone, the drug-resistant phenotype was obtained. BPIP did not inhibit the in vitro activity of recombinant BVDV RdRp, but did inhibit the activity of replication complexes (RCs). Computational docking revealed that F224 is located at the top of the finger domain of the polymerase. Docking of BPIP in the crystal structure of the BVDV RdRp revealed aromatic ring stacking, some hydrophobic contacts, and a hydrogen bond. Since two structurally unrelated compounds, i.e., BPIP and VP32947, target the same region of the BVDV RdRp, this position may be expected to be critical in the functioning of the polymerase or assembly of the RC. The potential of BPIP for the treatment of pestivirus and hepacivirus infections is discussed. PMID- 16352540 TI - VP2 cleavage and the leucine ring at the base of the fivefold cylinder control pH dependent externalization of both the VP1 N terminus and the genome of minute virus of mice. AB - Cylindrical projections surrounding the fivefold-symmetry axes in minute virus of mice (MVM) harbor central pores that penetrate through the virion shell. In newly released DNA-containing particles, these pores contain residues 28 to 38 belonging to a single copy of VP2, disposed so that its extreme N-terminal domain projects outside the particle. Virions are metastable, initially sequestering internally the N termini of all copies of the minor capsid protein, VP1, that is essential for entry. This VP1 domain can be externalized in vitro in response to limited heating, and we show here that the efficiency of this transition is greatly enhanced by proteolysis of VP2 N termini to yield VP3. This step also renders the VP1 rearrangement pH dependent, indicating that VP2 cleavage is a maturation step required to prime subsequent emergence of the VP1 "entry" domain. The tightest constriction within the cylinder is created by VP2 leucine 172, the five symmetry-related copies of which form a portal that resembles an iris diaphragm across the base of the pore. In MVMp, threonine substitution at this position, L172T, yields infectious particles following transfection at 37 degrees C, but these can initiate infection only at 32 degrees C, and this process can be blocked by exposing virions to a cellular factor(s) at 37 degrees C during the first 8 h after entry. At 32 degrees C, the mutant particle is highly infectious, and it remains stable prior to VP2 cleavage or following cleavage at pH 5.5 or below. However, upon exposure to neutral pH following VP2 cleavage, its VP1 specific sequences and genome are extruded even at room temperature, underscoring the significance of the VP2 cleavage step for MVM particle dynamics. PMID- 16352541 TI - Characterization of early steps in the poliovirus infection process: receptor decorated liposomes induce conversion of the virus to membrane-anchored entry intermediate particles. AB - The mechanism by which poliovirus infects the cell has been characterized by a combination of biochemical and structural studies, leading to a working model for cell entry. Upon receptor binding at physiological temperature, native virus (160S) undergoes a conformational change to a 135S particle from which VP4 and the N terminus of VP1 are externalized. These components interact with the membrane and are proposed to form a membrane pore. An additional conformational change in the particle is accompanied by release of the infectious viral RNA genome from the particle and its delivery, presumably through the membrane pore into the cytoplasm, leaving behind an empty 80S particle. In this report, we describe the generation of a receptor-decorated liposome system, comprising nickel-chelating nitrilotriacetic acid (NTA) liposomes and His-tagged poliovirus receptor, and its use in characterizing the early events in poliovirus infection. Receptor-decorated liposomes were able to capture virus and induce a temperature dependent virus conversion to the 135S particle. Upon conversion, 135S particles became tethered to the liposome independently of receptor by a membrane interaction with the N terminus of VP1. Converted particles had lost VP4, which partitioned with the membrane. The development of a simple model membrane system provides a novel tool for studying poliovirus entry. The liposome system bridges the gap between previous studies using either soluble receptor or whole cells and offers a flexible template which can be extrapolated to electron microscopy experiments that analyze the structural biology of nonenveloped virus entry. PMID- 16352542 TI - Human T-cell leukemia virus open reading frame II encodes a posttranscriptional repressor that is recruited at the level of transcription. AB - Human T-cell leukemia virus (HTLV) infection is a chronic, lifelong infection that is associated with the development of leukemia and neurological disease after a long latency period, and the mechanism by which the virus is able to evade host immune surveillance is elusive. Besides the structural and enzymatic proteins, HTLV encodes regulatory (Tax and Rex) and accessory (open reading frame I [ORF I] and ORF II) proteins. Tax activates viral and cellular transcription and promotes T-cell growth and malignant transformation. Rex acts posttranscriptionally to facilitate cytoplasmic expression of incompletely spliced viral mRNAs. Recently, we reported that the accessory gene products of HTLV-1 and HTLV-2 ORF II (p30II and p28II, respectively) are able to restrict viral replication. These proteins act as negative regulators of both Tax and Rex by binding to and retaining their mRNA in the nucleus, leading to reduced protein expression and virion production. Here, we show that p28II is recruited to the viral promoter in a Tax-dependent manner. After recruitment to the promoter, p28II or p30II then travels with the transcription elongation machinery until its target mRNA is synthesized. Experiments artificially directing these proteins to the promoter indicate that p28II, unlike HTLV-1 p30II, displays no transcriptional activity. Furthermore, the tethering of p28II directly to tax/rex mRNA resulted in repression of Tax function, which could be attributed to the ability of p28II to block TAP/p15-mediated enhancement of Tax expression. p28II mediated reduction of viral replication in infected cells may permit survival of the cells by allowing escape from immune recognition, which is consistent with the critical role of HTLV accessory proteins in viral persistence in vivo. PMID- 16352543 TI - Gamma interferon blocks gammaherpesvirus reactivation from latency. AB - Establishment of latent infection and reactivation from latency are critical aspects of herpesvirus infection and pathogenesis. Interfering with either of these steps in the herpesvirus life cycle may offer a novel strategy for controlling herpesvirus infection and associated disease pathogenesis. Prior studies show that mice deficient in gamma interferon (IFN-gamma) or the IFN-gamma receptor have elevated numbers of cells reactivating from murine gammaherpesvirus 68 (gammaHV68) latency, produce infectious virus after the establishment of latency, and develop large-vessel vasculitis. Here, we demonstrate that IFN-gamma is a powerful inhibitor of reactivation of gammaHV68 from latency in tissue culture. In vivo, IFN-gamma controls viral gene expression during latency. Importantly, depletion of IFN-gamma in latently infected mice results in an increased frequency of cells reactivating virus. This demonstrates that IFN-gamma is important for immune surveillance that limits reactivation of gammaHV68 from latency. PMID- 16352544 TI - The pseudorabies virus VP1/2 tegument protein is required for intracellular capsid transport. AB - Transport of capsids in cells is critical to alphaherpesvirus infection and pathogenesis; however, viral factors required for transport have yet to be identified. Here we provide a detailed examination of capsid dynamics during the egress phase of infection in Vero cells infected with pseudorabies virus. We demonstrate that the VP1/2 tegument protein is required for processive microtubule-based transport of capsids in the cytoplasm. A second tegument protein that binds to VP1/2, UL37, was necessary for wild-type transport but was not essential for this process. Both proteins were also required for efficient nuclear egress of capsids to the cytoplasm. PMID- 16352545 TI - Severe acute respiratory syndrome coronavirus 3a protein is released in membranous structures from 3a protein-expressing cells and infected cells. AB - Severe acute respiratory syndrome coronavirus (SCoV) accessory protein 3a is a virus structural protein. We demonstrate here that 3a protein was released efficiently in membranous structures from various cell lines expressing 3a protein. A subpopulation of the released 3a protein is associated with detergent resistant membranes. The presence of the YxxPhi and diacidic motifs, located within the cytoplasmic tail of the 3a protein, was not required for its efficient release. Analysis of supernatant from SCoV-infected cells with sucrose gradient sedimentation and virus capture assay indicated that the 3a protein was released from infected cells in two distinct populations, as a component of SCoV particles, and in membrane structures with a lower buoyant density. These data provide new insights into the biological properties of SCoV 3a protein. PMID- 16352546 TI - ICP0 prevents RNase L-independent rRNA cleavage in herpes simplex virus type 1 infected cells. AB - The classical interferon (IFN)-dependent antiviral response to viral infection involves the regulation of IFN-stimulated genes (ISGs), one being the gene encoding cellular endoribonuclease RNase L, which arrests protein synthesis and induces apoptosis by nonspecifically cleaving rRNA. Recently, the herpes simplex virus type 1 (HSV-1) protein ICP0 has been shown to block the induction of ISGs by subverting the IFN pathway upstream of the 2'-5'-oligoadenylate synthetase (OAS)/RNase L pathway. We report that ICP0 also prevents rRNA degradation at late stages of HSV-1 infection, independent of its E3 ubiquitin ligase activity, and that the resultant rRNA degradation is independent of the classical RNase L antiviral pathway. Moreover, the degradation is independent of the viral RNase vhs and is independent of IFN response factor 3. These studies indicate the existence of another, previously unidentified, RNase that is part of the host antiviral response to viral infection. PMID- 16352547 TI - Identification of a novel pathway essential for the immediate-early, interferon independent antiviral response to enveloped virions. AB - Viral infection elicits the activation of numerous cellular signal transduction pathways, leading to the induction of both innate and adaptive immunity. Previously we showed that entry of virion particles from a diverse array of enveloped virus families was capable of eliciting an interferon regulatory factor 3 (IRF-3)-mediated antiviral state in human fibroblasts in the absence of interferon production. Here we show that extracellular regulated kinase 1/2, p38 mitogen-activated protein kinase, and Jun N-terminal kinase/stress-activated protein kinase activities are not required for antiviral state induction. In contrast, treatment of cells with LY294002, an inhibitor of the phosphoinositide 3-kinase (PI3 kinase) family, prevents the induction of interferon-stimulated gene 56 (ISG56) and an antiviral response upon entry of virus particles. However, the prototypic class I p85/p110 PI3 kinase and its downstream effector Akt/PKB are dispensable for ISG and antiviral state induction. Furthermore, DNA-PK and PAK1, LY294002-sensitive members of the PI3 kinase family shown previously to be involved in IRF-3 activation, are also dispensable for ISG and antiviral state induction. The LY294002 inhibitor fails to prevent IRF-3 homodimerization or nuclear translocation upon virus particle entry. Together, these data suggest that virus entry triggers an innate antiviral response that requires the activity of a novel PI3 kinase family member. PMID- 16352548 TI - Changes in soluble factor-mediated CD8+ cell-derived antiviral activity in cynomolgus macaques infected with simian immunodeficiency virus SIVmac251: relationship to biological markers of progression. AB - Cross-sectional studies have shown that the capacity of CD8+ cells from human immunodeficiency virus (HIV)-infected patients and simian immunodeficiency virus (SIV) SIVmac-infected macaques to suppress the replication of human and simian immunodeficiency viruses in vitro depends on the clinical stage of disease, but little is known about changes in this antiviral activity over time in individual HIV-infected patients or SIV-infected macaques. We assessed changes in the soluble factor-mediated noncytolytic antiviral activity of CD8+ cells over time in eight cynomolgus macaques infected with SIVmac251 to determine the pathophysiological role of this activity. CD8+ cell-associated antiviral activity increased rapidly in the first week after viral inoculation and remained detectable during the early phase of infection. The net increase in antiviral activity of CD8+ cells was correlated with plasma viral load throughout the 15 months of follow-up. CD8+ cells gradually lost their antiviral activity over time and acquired virus replication-enhancing capacity. Levels of antiviral activity correlated with CD4+ T-cell counts after viral set point. Concentrations of beta chemokines and interleukin-16 in CD8+ cell supernatants were not correlated with this antiviral activity, and alpha-defensins were not detected. The soluble factor-mediated antiviral activity of CD8+ cells was neither cytolytic nor restricted to major histocompatibility complex. This longitudinal study strongly suggests that the increase in noncytolytic antiviral activity from baseline and the maintenance of this increase over time in cynomolgus macaques depend on both viral replication and CD4+ T cells. PMID- 16352549 TI - Host deadenylation-dependent mRNA decapping factors are required for a key step in brome mosaic virus RNA replication. AB - The genomes of positive-strand RNA [+RNA] viruses perform two mutually exclusive functions: they act as mRNAs for the translation of viral proteins and as templates for viral replication. A universal key step in the replication of +RNA viruses is the coordinated transition of the RNA genome from the cellular translation machinery to the viral replication complex. While host factors are involved in this step, their nature is largely unknown. By using the ability of the higher eukaryotic +RNA virus brome mosaic virus (BMV) to replicate in yeast, we previously showed that the host Lsm1p protein is required for efficient recruitment of BMV RNA from translation to replication. Here we show that in addition to Lsm1p, all tested components of the Lsm1p-7p/Pat1p/Dhh1p decapping activator complex, which functions in deadenylation-dependent decapping of cellular mRNAs, are required for BMV RNA recruitment for RNA replication. In contrast, other proteins of the decapping machinery, such as Edc1p and Edc2p from the deadenylation-dependent decapping pathway and Upf1p, Upf2p, and Upf3p from the deadenylation-independent decapping pathway, had no significant effects. The dependence of BMV RNA recruitment on the Lsm1p-7p/Pat1p/Dhh1p complex was linked exclusively to the 3' noncoding region of the BMV RNA. Collectively, our results suggest that the Lsm1p-7p/Pat1p/Dhh1p complex that transfers cellular mRNAs from translation to degradation might act as a key regulator in the switch from BMV RNA translation to replication. PMID- 16352550 TI - Defective assembly of influenza A virus due to a mutation in the polymerase subunit PA. AB - The RNA-dependent RNA polymerase of influenza A virus is composed of three subunits that together synthesize all viral mRNAs and also replicate the viral genomic RNA segments (vRNAs) through intermediates known as cRNAs. Here we describe functional characterization of 16 site-directed mutants of one polymerase subunit, termed PA. In accord with earlier studies, these mutants exhibited diverse, mainly quantitative impairments in expressing one or more classes of viral RNA, with associated infectivity defects of varying severity. One PA mutant, however, targeting residues 507 and 508, caused only modest perturbations of RNA expression yet completely eliminated the formation of plaque forming virus. Polymerases incorporating this mutant, designated J10, proved capable of synthesizing translationally active mRNAs and of replicating diverse cRNA or vRNA templates at levels compatible with viral infectivity. Both the mutant protein and its RNA products were appropriately localized in the cytoplasm, where influenza virus assembly occurs. Nevertheless, J10 failed to generate infectious particles from cells in a plasmid-based influenza virus assembly assay, and hemagglutinating material from the supernatants of such cells contained little or no nuclease-resistant genomic RNA. These findings suggest that PA has a previously unrecognized role in assembly or release of influenza virus virions, perhaps influencing core structure or the packaging of vRNAs or other essential components into nascent influenza virus particles. PMID- 16352551 TI - Identification and characterization of a Penaeus monodon lymphoid cell-expressed receptor for the yellow head virus. AB - The yellow head virus is a positive-sense, single-stranded RNA virus that causes significant mortality in farmed penaeid shrimp. This study sought to isolate and characterize the receptor protein used by the virus to gain entry into Penaeus monodon Oka (lymphoid) organ cells, a primary target of yellow head virus infections. Virus overlay protein binding assay on Oka organ membrane preparations identified a 65-kDa protein, and antibodies raised against this protein inhibited virus entry in primary Oka cell cultures by approximately 80%. N-terminal sequence analysis of the 65-kDa protein generated a 17-amino acid peptide fragment which was used to design degenerate primers that amplified a 1.5 kbp product from Oka organ total RNA, which was cloned and sequenced. Northern analysis and PCR were used to confirm a single RNA transcript that was expressed in most tissues. Subsequently, the mature cDNA was recloned and the expressed protein shown to cross-react with the antibody raised against the original virus binding band. Down regulation of the message through double-stranded RNA-mediated RNA interference silencing resulted in the complete inhibition of virus entry. While the identity of the clone remains unknown, it nevertheless represents the first invertebrate Nidovirus receptor isolated to date. PMID- 16352552 TI - Lack of intrinsic CTLA-4 expression has minimal effect on regulation of antiviral T-cell immunity. AB - CTLA-4 is considered one of the most potent negative regulators of T-cell activation. To circumvent experimental limitations due to fatal lymphoproliferative disease associated with genetic ablation of CTLA-4, we have used radiation chimeras reconstituted with a mixture of CTLA-4+/+ and CTLA-4-/- bone marrow that retain a normal phenotype and allow the evaluation of long-term T-cell immunity under conditions of intrinsic CTLA-4 deficiency. Following virus infection, we profiled primary, memory, and secondary CD8+ and CD4+ T-cell responses directed against eight different viral epitopes. Our data demonstrate unaltered antigen-driven proliferation, acquisition of effector functions, distribution of epitope hierarchies, T-cell receptor repertoire selection, functional avidities, and long-term memory maintenance in the absence of CTLA-4. Moreover, regulation of memory T-cell survival and homeostatic proliferation, as well as secondary responses, was equivalent in virus-specific CTLA4+/+ and CTL-A 4-/- T-cell populations. Thus, lack of CTLA-4 expression by antigen-specific T cells can be compensated for by extrinsic factors in the presence of CTLA-4 expression by other cells. These findings have implications for the physiologic, pathological, and therapeutic regulation of costimulation. PMID- 16352553 TI - Mechanism of telomerase activation by v-Rel and its contribution to transformation. AB - Telomerase is activated during the transformation of lymphoid cells and fibroblasts by v-Rel, the oncogenic member of the Rel/NF-kappaB family of transcription factors. v-Rel-transformed cell lines have longer telomeres than untransformed chicken lymphoid cells and have high levels of telomerase activity. v-Rel-mediated activation of telomerase is achieved by multiple mechanisms. The expression of the gene encoding the catalytic subunit of telomerase (TERT) was directly upregulated by v-Rel. Moreover, the expression of v-Rel altered the ratio of alternatively spliced and full-length TERT transcripts in favor of the full-length forms. The activation of telomerase by v-Rel in lymphocytes was also accompanied by inactivation of nuclear inhibitors. The inhibition of telomerase activity in v-Rel-transformed cell lines led to apoptosis within 24 h. The expression of v-Rel in a macrophage cell line resulted in elevated levels of reactive oxygen species (ROS), increased telomerase activity, and increased sensitivity to telomerase inhibitors. In contrast, the ectopic expression of TERT decreased the extent of apoptosis induced by ROS. The activation of telomerase by v-Rel may, therefore, partially protect the transformed cells from apoptosis induced by ROS. PMID- 16352554 TI - Neither the RNA nor the proteins of open reading frames 3a and 3b of the coronavirus infectious bronchitis virus are essential for replication. AB - Gene 3 of infectious bronchitis virus is tricistronic; open reading frames (ORFs) 3a and 3b encode two small nonstructural (ns) proteins, 3a and 3b, of unknown function, and a third, structural protein E, is encoded by ORF 3c. To determine if either the 3a or the 3b protein is required for replication, we first modified their translation initiation codons to prevent translation of the 3a and 3b proteins from recombinant infectious bronchitis viruses (rIBVs). Replication in primary chick kidney (CK) cells and in chicken embryos was not affected. In chicken tracheal organ cultures (TOCs), the recombinant rIBVs reached titers similar to those of the wild-type virus, but in the case of viruses lacking the 3a protein, the titer declined reproducibly earlier. Translation of the IBV E protein is believed to be initiated by internal entry of ribosomes at a structure formed by the sequences corresponding to ORFs 3a and 3b. To assess the necessity of this mechanism, we deleted most of the sequence representing 3a and 3b to produce a gene in which ORF 3c (E) was adjacent to the gene 3 transcription associated sequence. Western blot analysis revealed that the recombinant IBV produced fivefold less E protein. Nevertheless, titers produced in CK cells, embryos, and TOCs were similar to those of the wild-type virus, although they declined earlier in TOCs, probably due to the absence of the 3a protein. Thus, neither the tricistronic arrangement of gene 3, the internal initiation of translation of E protein, nor the 3a and 3b proteins are essential for replication per se, suggesting that these proteins are accessory proteins that may have roles in vivo. PMID- 16352555 TI - Characterization of the vaccinia virus A35R protein and its role in virulence. AB - The vaccinia virus A35R gene is highly conserved among poxviruses and encodes a previously uncharacterized hydrophobic acidic protein. Western blotting with anti A35R peptide antibodies indicated that the protein is expressed early in infection and resolved as a single sharp band of approximately 23 kDa, slightly higher than the 20 kDa predicted from its sequence. The protein band appeared to be the same molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whether expressed in an in vitro transcription/translation system without microsomes or expressed in infected cells, suggesting that it was not glycosylated. A mutant virus with the A35R gene deleted (vA35Delta) formed wild-type-sized plaques on all cell lines tested (human, monkey, mouse, and rabbit); thus, A35R is not required for replication and does not appear to be a host range gene. Although the A35R protein is hydrophobic, it is unlikely to be an integral membrane protein, as it partitioned to the aqueous phase during TX 114 partitioning. The protein could not be detected in virus-infected cell supernatants. A35R localized intracellularly to the virus factories, where the first stages of morphogenesis occur. The vA35Delta mutant formed near-normal levels of the various morphogenic stages of infectious virus particles and supported normal acid-induced fusion of virus-infected cells. Despite normal growth and morphogenesis in vitro, the vA35Delta mutant virus was attenuated in intranasal challenge of mice compared to wild-type and A35R rescue virus. Thus, the intracellular A35R protein plays a role in virulence. The A35R has little homology to any protein outside of poxviruses, suggesting a novel virulence mechanism. PMID- 16352556 TI - Characterization of mimivirus DNA topoisomerase IB suggests horizontal gene transfer between eukaryal viruses and bacteria. AB - Mimivirus, a parasite of Acanthamoeba polyphaga, is the largest DNA virus known; it encodes dozens of proteins with imputed functions in nucleic acid transactions. Here we produced, purified, and characterized mimivirus DNA topoisomerase IB (TopIB), which we find to be a structural and functional homolog of poxvirus TopIB and the poxvirus-like topoisomerases discovered recently in bacteria. Arginine, histidine, and tyrosine side chains responsible for TopIB transesterification are conserved and essential in mimivirus TopIB. Moreover, mimivirus TopIB is capable of incising duplex DNA at the 5'-CCCTT cleavage site recognized by all poxvirus topoisomerases. Based on the available data, mimivirus TopIB appears functionally more akin to poxvirus TopIB than bacterial TopIB, despite its greater primary structure similarity to the bacterial TopIB group. We speculate that the ancestral bacterial/viral TopIB was disseminated by horizontal gene transfer within amoebae, which are permissive hosts for either intracellular growth or persistence of many present-day bacterial species that have a type IB topoisomerase. PMID- 16352557 TI - Inactivation of prions by acidic sodium dodecyl sulfate. AB - Prompted by the discovery that prions become protease-sensitive after exposure to branched polyamine dendrimers in acetic acid (AcOH) (S. Supattapone, H. Wille, L. Uyechi, J. Safar, P. Tremblay, F. C. Szoka, F. E. Cohen, S. B. Prusiner, and M. R. Scott, J. Virol. 75:3453-3461, 2001), we investigated the inactivation of prions by sodium dodecyl sulfate (SDS) in weak acid. As judged by sensitivity to proteolytic digestion, the disease-causing prion protein (PrPSc) was denatured at room temperature by SDS at pH values of < or =4.5 or > or =10. Exposure of Sc237 prions in Syrian hamster brain homogenates to 1% SDS and 0.5% AcOH at room temperature resulted in a reduction of prion titer by a factor of ca. 10(7); however, all of the bioassay hamsters eventually developed prion disease. When various concentrations of SDS and AcOH were tested, the duration and temperature of exposure acted synergistically to inactivate both hamster Sc237 prions and human sporadic Creutzfeldt-Jakob disease (sCJD) prions. The inactivation of prions in brain homogenates and those bound to stainless steel wires was evaluated by using bioassays in transgenic mice. sCJD prions were more than 100,000 times more resistant to inactivation than Sc237 prions, demonstrating that inactivation procedures validated on rodent prions cannot be extrapolated to inactivation of human prions. Some procedures that significantly reduced prion titers in brain homogenates had a limited effect on prions bound to the surface of stainless steel wires. Using acidic SDS combined with autoclaving for 15 min, human sCJD prions bound to stainless steel wires were eliminated. Our findings form the basis for a noncorrosive system that is suitable for inactivating prions on surgical instruments, as well as on other medical and dental equipment. PMID- 16352558 TI - In vivo and in vitro analysis of factor binding sites in Jaagsiekte sheep retrovirus long terminal repeat enhancer sequences: roles of HNF-3, NF-I, and C/EBP for activity in lung epithelial cells. AB - Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma, a contagious lung cancer of sheep that arises from type II pneumocytes and Clara cells of the lung epithelium. Studies of the tropism of this virus have been hindered by the lack of an efficient system for viral replication in tissue culture. To map regulatory regions important for transcriptional activation, an in vivo footprinting method that couples dimethyl sulfate treatment and ligation-mediated PCR was performed in murine type II pneumocyte-derived MLE-15 cells infected with a chimeric Moloney murine leukemia virus driven by the JSRV enhancers (DeltaMo+JS Mo-MuLV). In vivo footprints were found in the JSRV enhancers in two regions previously shown to be important for JSRV long terminal repeat (LTR) activity: a binding site for the lung-specific transcription factor HNF-3beta and an E-box element in the distal enhancer adjacent to an NF-kappaB-like binding site. In addition, in vivo footprints were detected in two downstream motifs likely to bind C/EBP and NF-I. Mutational analysis of a JSRV LTR reporter construct (pJS21luc) revealed that the C/EBP binding site is critical for LTR activity, while the putative NF-I binding element is less important; elimination of these sites resulted in 70% and 40% drops in LTR activity, respectively. Electrophoretic mobility shift assays using nuclear extracts from MLE-15 murine Clara cell-derived mtCC1-2 cells with probes corresponding to the NF-I or C/EBP sites revealed several complexes. Antiserum directed against NF-IA, C/EBPalpha, or C/EBPbeta supershifted the corresponding protein-DNA complexes, indicating that these isoforms, which are also important for the expression of several cellular lung-specific genes, may be important for JSRV expression in lung epithelial cells. PMID- 16352559 TI - Interaction of moloney murine leukemia virus capsid with Ubc9 and PIASy mediates SUMO-1 addition required early in infection. AB - Yeast two-hybrid screens led to the identification of Ubc9 and PIASy, the E2 and E3 small ubiquitin-like modifier (SUMO)-conjugating enzymes, as proteins interacting with the capsid (CA) protein of the Moloney murine leukemia virus. The binding site in CA for Ubc9 was mapped by deletion and alanine-scanning mutagenesis to a consensus motif for SUMOylation at residues 202 to 220, and the binding site for PIASy was mapped to residues 114 to 176, directly centered on the major homology region. Expression of CA and a tagged SUMO-1 protein resulted in covalent transfer of SUMO-1 to CA in vivo. Mutations of lysine residues to arginines near the Ubc9 binding site and mutations at the PIASy binding site reduced or eliminated CA SUMOylation. Introduction of these mutations into the complete viral genome blocked virus replication. The mutants exhibited no defects in the late stages of viral gene expression or virion assembly. Upon infection, the mutant viruses were able to carry out reverse transcription to synthesize normal levels of linear viral DNA but were unable to produce the circular viral DNAs or integrated provirus normally found in the nucleus. The results suggest that the SUMOylation of CA mediated by an interaction with Ubc9 and PIASy is required for early events of infection, after reverse transcription and before nuclear entry and viral DNA integration. PMID- 16352560 TI - Expanded tropism and altered activation of a retroviral glycoprotein resistant to an entry inhibitor peptide. AB - The envelope of class I viruses can be a target for potent viral inhibitors, such as the human immunodeficiency virus type 1 (HIV-1) inhibitor enfuvirtide, which are derived from the C-terminal heptad repeat (HR2) of the transmembrane (TM) subunit. Resistance to an HR2-based peptide inhibitor of a model retrovirus, subgroup A of the Avian Sarcoma and Leukosis Virus genus (ASLV-A), was studied by examining mutants derived by viral passage in the presence of inhibitor. Variants with reduced sensitivity to inhibitor were readily selected in vitro. Sensitivity determinants were identified for 13 different isolates, all of which mapped to the TM subunit. These determinants were identified in two regions: (i) the N terminal heptad repeat (HR1) and (ii) the N-terminal segment of TM, between the subunit cleavage site and the fusion peptide. The latter class of mutants identified a region outside of the predicted HR2-binding site that can significantly alter sensitivity to inhibitor. A subset of the HR1 mutants displayed the unanticipated ability to infect nonavian cells. This expanded tropism was associated with increased efficiency of envelope triggering by soluble receptor at low temperatures, as measured by protease sensitivity of the surface subunit (SU) of envelope. In addition, expanded tropism was linked for the most readily triggered mutants with increased sensitivity to neutralization by SU-specific antiserum. These observations depict a class of HR2 peptide selected mutations with a reduced activation threshold, thereby allowing the utilization of alternative receptors for viral entry. PMID- 16352561 TI - Role for nsP2 proteins in the cessation of alphavirus minus-strand synthesis by host cells. AB - In order to establish nonlytic persistent infections (PI) of BHK cells, replicons derived from Sindbis (SIN) and Semliki Forest (SFV) viruses have mutations in nsP2. Five different nsP2 PI replicons were compared to wild-type (wt) SIN, SFV, and wt nsPs SIN replicons. Replicon PI BHK21 cells had viral RNA synthesis rates that were less than 5% of those of the wt virus and approximately 10% or less of those of SIN wt replicon-infected cells, and, in contrast to wt virus and replicons containing wt nsP2, all showed a phenotype of continuous minus-strand synthesis and of unstable, mature replication/transcription complexes (RC+) that are active in plus-strand synthesis. Minus-strand synthesis and incorporation of [3H]uridine into replicative intermediates differed among PI replicons, depending on the location of the mutation in nsP2. Minus-strand synthesis by PI cells appeared normal; it was dependent on continuous P123 and P1234 polyprotein synthesis and ceased when protein synthesis was inhibited. The failure by the PI replicons to shut off minus-strand synthesis was not due to some defect in the PI cells but rather was due to the loss of some function in the mutated nsP2. This was demonstrated by showing that superinfection of PI cells with wt SFV triggered the shutdown of minus-strand synthesis, which we believe is a host response to infection with alphaviruses. Together, the results indicate alphavirus nsP2 functions to engage the host response to infection and activate a switch from the early-to-late phase. The loss of this function leads to continuous viral minus strand synthesis and the production of unstable RC+. PMID- 16352562 TI - Cytokine responses in gnotobiotic pigs after infection with virulent or attenuated human rotavirus. AB - To understand the role of cytokines during rotavirus infection, we assessed the kinetics of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) (proinflammatory), IL-12 (Th1 inducer), gamma interferon (IFN-gamma) (Th1), IL-4 and IL-10 (Th2), and transforming growth factor beta (Th3) cytokine responses by enzyme-linked immunosorbent assay in serum and intestinal contents of neonatal gnotobiotic pigs and IL-12, IFN-gamma, IL-4, and IL-10 cytokine-secreting cell (CSC) responses of mononuclear cells from ileum, spleen, and blood by ELISPOT. Pigs received the virulent Wa P1A[8]G1 strain of human rotavirus (HRV) (VirHRV), attenuated Wa HRV (AttHRV), or mock (controls). The TNF-alpha levels peaked earlier and remained elevated in serum of the VirHRV group but peaked later in the AttHRV group. In serum, IL-6 was significantly elevated at postinoculation day (PID) 1 in the VirHRV group and at PID 3 in both HRV groups. The IL-12 was detected in serum of all pigs including controls with significantly elevated peaks in both HRV-infected groups, indicating a role for IL-12 in the induction of immune responses to rotavirus infection. Only low and transient IFN-gamma responses occurred in serum and intestinal contents of the AttHRV-infected pigs, compared to significantly higher and prolonged IFN-gamma responses in the VirHRV infected pigs. This observation coincides with the diarrhea and viremia induced by VirHRV. The number of IFN-gamma-secreting cells was significantly higher in the ileum of the VirHRV group than in that of the controls. The number of IL-4 CSCs was significantly higher in ileum of both HRV groups than in that of the controls. Significantly higher levels of IL-10 in the serum occurred early in the VirHRV group, compared to lower levels in the AttHRV group. However, the number of IL-10 CSCs was significantly higher later in ileum and spleen of the AttHRV than in the VirHRV group, suggesting a delayed initiation of a Th2 response induced by AttHRV. A significantly higher percentage of pigs had IFN-gamma and IL 10 responses in serum after VirHRV infection than after AttHRV infection or in controls. These data indicate a balanced Th1/Th2 response during rotavirus infection, with higher cytokine levels early after infection with VirHRV compared to that with AttHRV. Mapping the kinetics and patterns of cytokine responses after rotavirus infection has important implications for induction of protective immunity by HRV vaccines. Higher protection rates may be associated with more balanced Th1- and Th2-type responses, but induction of higher earlier IFN-gamma (Th1) and proinflammatory cytokines triggered by VirHRV may also play an important role in the higher intestinal immunoglobulin A responses and protection rates induced by VirHRV. PMID- 16352563 TI - Prostate tumor cells infected with a recombinant influenza virus expressing a truncated NS1 protein activate cytolytic CD8+ cells to recognize noninfected tumor cells. AB - Many viral oncolytic approaches against cancer are based on the ability of specific viruses to replicate in tumors expressing components of the constitutively activated Ras/mitogen-activated protein kinase (MAPK) pathways and/or inhibited or dysregulated alpha/beta interferon (IFN-alpha/beta) response pathways. A major issue when considering these approaches is their applicability to tumors that lack activated Ras. To identify the effector mechanisms activated by oncolytic viruses, we investigated inhibition of proliferation of the prostate cancer line LNCap by the recombinant TR-NS1 influenza A virus, a genetically attenuated influenza A/PR8/34 virus expressing a truncated nonstructural protein (NS1) of 126 amino acids. LNCap cells lack constitutively activated MAPK, extracellular signal-regulated kinase (ERK), and p38 and are resistant to death by IFN-alpha. Truncation of the NS1 protein of influenza viruses is known to result in viral attenuation due to a reduced ability of the NS1 to inhibit the IFN-alpha/beta response. Infection with TR-NS1 virus rapidly activated ERK-1 more than ERK-2 in LNCap cells. Importantly, TR-NS1 virus infection transiently inhibited cell proliferation and induced apoptosis in LNCap cells. Addition of peripheral blood mononuclear cells (PBMC) and interleukin 12 (IL-12) to TR-NS1 virus-infected LNCap cells (TR-NS1-LNCap) resulted in faster elimination of TR NS1-LNCap cells compared with LNCap cells. Moreover, TR-NS1-LNCap cells induced IFN-gamma in PBMC. The levels of IFN-gamma were amplified by IL-12. TR-NS1-LNCap cells also induced tumor-lytic cytotoxic T lymphocytes (CTL). These CTL lysed noninfected LNCap cells in a CD8-dependent manner. Activation of cellular immunity to tumor cells by viruses is an intriguing effector pathway, which should be especially significant for elimination of human tumors that lack activated Ras. PMID- 16352564 TI - Role of the mitochondrial signaling pathway in murine coronavirus-induced oligodendrocyte apoptosis. AB - A previous study demonstrated that infection of rat oligodendrocytes by mouse hepatitis virus (MHV) resulted in apoptosis, which is caspase dependent (Y. Liu, Y. Cai, and X. Zhang, J. Virol. 77:11952-11963, 2003). Here we determined the involvement of the mitochondrial pathway in MHV-induced oligodendrocyte apoptosis. We found that caspase-9 activity was 12-fold higher in virus-infected cells than in mock-infected cells at 24 h postinfection (p.i.). Pretreatment of cells with a caspase-9 inhibitor completely blocked caspase-9 activation and partially inhibited the apoptosis mediated by MHV infection. Analyses of cytochrome c release further revealed an activation of the mitochondrial apoptotic pathway. Stable overexpression of the two antiapoptotic proteins Bcl-2 and Bcl-xL significantly, though only partially, blocked apoptosis, suggesting that activation of the mitochondrial pathway is partially responsible for the apoptosis. To identify upstream signals, we determined caspase-8 activity, cleavage of Bid, and expression of Bax and Bad by Western blotting. We found a drastic increase in caspase-8 activity and cleavage of Bid at 24 h p.i. in virus infected cells, suggesting that Bid may serve as a messenger to relay the signals from caspase-8 to mitochondria. However, treatment with a caspase-8 inhibitor only slightly blocked cytochrome c release from the mitochondria. Furthermore, we found that Bax but not Bad was significantly increased at 12 h p.i. in cells infected with both live and UV-inactivated viruses and that Bax activation was partially blocked by treatment with the caspase-8 inhibitor. These results thus establish the involvement of the mitochondrial pathway in MHV-induced oligodendrocyte apoptosis. PMID- 16352565 TI - Hepatitis C virus subgenomic replicon requires an active NS3 RNA helicase. AB - Mutations were introduced into the NS3 helicase region of a hepatitis C virus (HCV) Con1 subgenomic replicon to ascertain the role of the helicase in viral replication. One new replicon lacked two-thirds of the NS3 helicase (Deltahel), and six others contained one of the following six amino acid substitutions in NS3: R393A, F438A, T450I, E493K, W501A, and W501F. It has been previously reported that purified R393A, F438A, and W501A HCV helicase proteins do not unwind RNA but unwind DNA, bind RNA, and hydrolyze ATP. On the other hand, previous data suggest that a W501F protein retains most of its unwinding abilities and that purified T450I and E493K HCV helicase proteins have enhanced unwinding abilities. In a hepatoma cell line that has been cured of HCV replicons using interferon, the T450I and W501F replicons synthesized both negative-sense and positive-sense viral RNA and formed colonies after selection with similar efficiencies as the parent replicon. However, the Deltahel, R393A, F438A, and W501A replicons encoded and processed an HCV polyprotein but did not synthesize additional viral RNA or form colonies. Surprisingly the same phenotype was seen for the E493K replicon. The inability of the E493K replicon to replicate might point to a role of pH in viral replication because a previous analysis has shown that, unlike the wild-type NS3 protein, the helicase activity of an E493K protein is not sensitive to pH changes. These results demonstrate that the RNA-unwinding activity of the HCV NS3 helicase is needed for RNA replication. PMID- 16352566 TI - N- and C-terminal cooperation in rotavirus enterotoxin: novel mechanism of modulation of the properties of a multifunctional protein by a structurally and functionally overlapping conformational domain. AB - Rotavirus NSP4 is a multifunctional endoplasmic reticulum (ER)-resident nonstructural protein with the N terminus anchored in the ER and about 131 amino acids (aa) of the C-terminal tail (CT) oriented in the cytoplasm. Previous studies showed a peptide spanning aa 114 to 135 to induce diarrhea in newborn mouse pups with the 50% diarrheal dose approximately 100-fold higher than that for the full-length protein, suggesting a role for other regions in the protein in potentiating its diarrhea-inducing ability. In this report, employing a large number of methods and deletion and amino acid substitution mutants, we provide evidence for the cooperation between the extreme C terminus and a putative amphipathic alpha-helix located between aa 73 and 85 (AAH73-85) at the N terminus of DeltaN72, a mutant that lacked the N-terminal 72 aa of nonstructural protein 4 (NSP4) from Hg18 and SA11. Cooperation between the two termini appears to generate a unique conformational state, specifically recognized by thioflavin T, that promoted efficient multimerization of the oligomer into high-molecular-mass soluble complexes and dramatically enhanced resistance against trypsin digestion, enterotoxin activity of the diarrhea-inducing region (DIR), and double-layered particle-binding activity of the protein. Mutations in either the C terminus, AAH73-85, or the DIR resulted in severely compromised biological functions, suggesting that the properties of NSP4 are subject to modulation by a single and/or overlapping highly sensitive conformational domain that appears to encompass the entire CT. Our results provide for the first time, in the absence of a three-dimensional structure, a unique conformation-dependent mechanism for understanding the NSP4-mediated pleiotropic properties including virus virulence and morphogenesis. PMID- 16352567 TI - Liver transduction with recombinant adeno-associated virus is primarily restricted by capsid serotype not vector genotype. AB - We and others have recently reported highly efficient liver gene transfer with adeno-associated virus 8 (AAV-8) pseudotypes, i.e., AAV-2 genomes packaged into AAV-8 capsids. Here we studied whether liver transduction could be further enhanced by using viral DNA packaging sequences (inverted terminal repeats [ITRs]) derived from AAV genotypes other than 2. To this end, we generated two sets of vector constructs carrying expression cassettes embedding a gfp gene or the human factor IX (hfIX) gene flanked by ITRs from AAV genotypes 1 through 6. Initial in vitro analyses of gfp vector DNA replication, encapsidation, and cell transduction revealed a surprisingly high degree of interchangeability among the six genotypes. For subsequent in vivo studies, we cross-packaged the six hfIX variants into AAV-8 and infused mice via the portal vein with doses of 5 x 10(10) to 1.8 x 10(12) particles. Notably, all vectors expressed comparably high plasma hFIX levels within a dose cohort over the following 6 months, concurrent with the finding of equivalent vector DNA copy numbers per cell. Partial hepatectomies resulted in approximately 80% drops of hFIX levels and vector DNA copy numbers in all groups, indicating genotype-independent persistence of predominantly episomal vector DNA. Southern blot analyses of total liver DNA in fact confirmed the presence of identical and mostly nonintegrated molecular vector forms for all genotypes. We conclude that, unlike serotypes, AAV genotypes are not critical for efficient hepatocyte transduction and can be freely substituted. This corroborates our current model for AAV vector persistence in the liver and provides useful information for the future design and application of recombinant AAV. PMID- 16352568 TI - Point mutations in herpes simplex virus type 1 oriL, but not in oriS, reduce pathogenesis during acute infection of mice and impair reactivation from latency. AB - In vitro studies of herpes simplex virus type 1 (HSV-1) viruses containing mutations in core sequences of the viral origins of DNA replication, oriL and oriS, that eliminate the ability of these origins to initiate viral-DNA synthesis have demonstrated little or no effect on viral replication in cultured cells, leading to the conclusion that the two types of origins are functionally redundant. It remains unclear, therefore, why origins that appear to be redundant are maintained evolutionarily in HSV-1 and other neurotropic alphaherpesviruses. To test the hypothesis that oriL and oriS have distinct functions in the HSV-1 life cycle in vivo, we determined the in vivo phenotypes of two mutant viruses, DoriL-I(LR) and DoriS-I, containing point mutations in oriL and oriS site I, respectively, that eliminate origin DNA initiation function. Following corneal inoculation of mice, tear film titers of DoriS-I were reduced relative to wild type virus. In all other tests, however, DoriS-I behaved like wild-type virus. In contrast, titers of DoriL-I(LR) in tear film, trigeminal ganglia (TG), and hindbrain were reduced and mice infected with DoriL-I(LR) exhibited greatly reduced mortality relative to wild-type virus. In the TG explant and TG cell culture models of reactivation, DoriL-I(LR) reactivated with delayed kinetics and, in the latter model, with reduced efficiency relative to wild-type virus. Rescuant viruses DoriL-I(LR)-R and DoriS-I-R behaved like wild-type virus in all tests. These findings demonstrate that functional differences exist between oriL and oriS and reveal a prominent role for oriL in HSV-1 pathogenesis. PMID- 16352569 TI - Mutations in the U5 sequences adjacent to the primer binding site do not affect tRNA cleavage by rous sarcoma virus RNase H but do cause aberrant integrations in vivo. AB - In most retroviruses, the first nucleotide added to the tRNA primer becomes the right end of the U5 region in the right long terminal repeat (LTR); the removal of this tRNA primer by RNase H defines the right end of the linear double stranded DNA. Most retroviruses have two nucleotides between the 5' end of the primer binding site (PBS) and the CA dinucleotide that will become the end of the integrated provirus. However, human immunodeficiency virus type 1 (HIV-1) has only one nucleotide at this position, and HIV-2 has three nucleotides. We changed the two nucleotides (TT) between the PBS and the CA dinucleotide of the Rous sarcoma virus (RSV)-derived vector RSVP(A)Z to match the HIV-1 sequence (G) and the HIV-2 sequence (GGT), and we changed the CA dinucleotide to TC. In all three mutants, RNase H removes the entire tRNA primer. Sequence analysis of RSVP(HIV2) proviruses suggests that RSV integrase can remove three nucleotides from the U5 LTR terminus of the linear viral DNA during integration, although this mutation significantly reduced virus titer, suggesting that removing three nucleotides is inefficient. However, the results obtained with RSVP(HIV1) and RSVP(CATC) show that RSV integrase can process and integrate the normal U3 LTR terminus of a linear DNA independently of an aberrant U5 LTR terminus. The aberrant end can then be joined to the host DNA by unusual processes that do not involve the conserved CA dinucleotide. These unusual events generate either large duplications or, less frequently, deletions in the host genomic DNA instead of the normal 5- to 6-base duplications. PMID- 16352570 TI - Nonstructural protein 3 of bluetongue virus assists virus release by recruiting ESCRT-I protein Tsg101. AB - The release of Bluetongue virus (BTV) and other members of the Orbivirus genus from infected host cells occurs predominantly by cell lysis, and in some cases, by budding from the plasma membrane. Two nonstructural proteins, NS3 and NS3A, have been implicated in this process. Here we show that both proteins bind to human Tsg101 and its ortholog from Drosophila melanogaster with similar strengths in vitro. This interaction is mediated by a conserved PSAP motif in NS3 and appears to play a role in virus release. The depletion of Tsg101 with small interfering RNA inhibits the release of BTV and African horse sickness virus, a related orbivirus, from HeLa cells up to fivefold and threefold, respectively. Like most other viral proteins which recruit Tsg101, NS3 also harbors a PPXY late domain motif that allows NS3 to bind NEDD4-like ubiquitin ligases in vitro. However, the late-domain motifs in NS3 do not function as effectively in facilitating the release of mini Gag virus-like particles from 293T cells as the late domains from human immunodeficiency virus type 1, human T-cell leukemia virus, and Ebola virus. A mutagenesis study showed that the arginine residue in the PPRY motif is responsible for the low activity of the NS3 late-domain motifs. Our data suggest that the BTV late-domain motifs either recruit an antagonist that interferes with budding or fail to recruit an agonist which is different from NEDD4. PMID- 16352571 TI - Resistance of neonatal mice to scrapie is associated with inefficient infection of the immature spleen. AB - Previous studies demonstrated that neonatal mice up to about a week old are less susceptible than adult mice to infection by intraperitoneal inoculation with mouse-passaged scrapie. In peripherally inoculated adult mice, scrapie replicates in lymphoid tissues such as the spleen before invading the central nervous system. Here, we investigated scrapie susceptibility in neonatal mice in more detail, concentrating on spleen involvement. First, we demonstrated that neonatal mice are about 10 times less susceptible than adults to intraperitoneal scrapie inoculation. Then we injected mice intraperitoneally with a scrapie dose that produced disease in all mice inoculated at 10 days or older but in only about a third of neonatally inoculated mice. In this experiment, spleens collected 70 days after scrapie injection of mice 10 days old or older almost all contained pathological prion protein, PrPSc, and those that were bioassayed all contained high infectivity levels. In contrast, at this early stage, only two of six spleens from neonatally inoculated mice had detectable, low infectivity levels; no PrPSc was detected, even in the two spleens. Therefore, neonatal mice have an impaired ability to replicate scrapie in their spleens, suggesting that replication sites are absent or sparse at birth but mature within 10 days. The increase in susceptibility with age correlated with the first immunocytochemical detection of the normal cellular form of prion protein, PrPc, on maturing follicular dendritic cell networks. As lymphoid tissues are more mature at birth in sheep, cattle, and humans than in mice, our results suggest that in utero infection with scrapie-like agents is theoretically possible in these species. PMID- 16352572 TI - Frequency and dynamics of recombination within different species of human enteroviruses. AB - Enteroviruses are members of the family Picornaviridae that cause widespread infections in human and other mammalian populations. Enteroviruses are genetically and antigenically highly variable, and recombination within and between serotypes contributes to their genetic diversity. To investigate the dynamics of the recombination process, sequence phylogenies between three regions of the genome (VP4, VP1, and 3Dpol) were compared among species A and B enterovirus variants detected in a human population-based survey in Scotland between 2000 and 2001, along with contemporary virus isolates collected in the same geographical region. This analysis used novel bioinformatic methods to quantify phylogenetic compatibility and correlations with serotype assignments of evolutionary trees constructed for different regions of the enterovirus genome. Species B enteroviruses showed much more frequent, time-correlated recombination events than those found for species A, despite the equivalence in population sampling, concordant with a linkage analysis of previously characterized enterovirus sequences obtained over longer collection periods. An analysis of recombination among complete genome sequences by computation of a phylogenetic compatibility matrix (PCM) demonstrated sharply defined boundaries between the VP2/VP3/VP1 block and sequences to either side in phylogenetic compatibility. The PCM also revealed equivalent or frequently greater degrees of incompatibility between different parts within the nonstructural region (2A-3D), indicating the occurrence of extensive recombination events in the past evolution of this part of the genome. Together, these findings provide new insights into the dynamics of species A and B enterovirus recombination and evolution and into the contribution of structured sampling to documenting reservoirs, emergence, and spread of novel recombinant forms in human populations. PMID- 16352573 TI - Herpes simplex virus type 1 infection induces activation and recruitment of protein kinase C to the nuclear membrane and increased phosphorylation of lamin B. AB - We report that herpes simplex virus type 1 (HSV-1) infection leads to the recruitment of protein kinase C (PKC) to the nuclear rim. In HEp-2 cells, PKC recruitment to the nuclear rim was initiated between 8 h and 12 h postinfection. PKCdelta, a proapoptotic kinase, was completely recruited to the nuclear rim upon infection with HSV-1. PKCalpha was less dramatically relocalized mostly at the nuclear rim upon infection, although some PKCalpha remained in the cytoplasm. PKCzeta-specific immunofluorescence was not significantly relocated to the nuclear rim. The UL34 and UL31 proteins, as well as their association, were each required for PKC recruitment to the nuclear rim. The HSV-1 US3 protein product, a kinase which regulates the phosphorylation state and localization of UL34, was not required for PKC recruitment to the nuclear rim; however, it was required for proper localization along the nuclear rim, as PKC appeared unevenly distributed along the nuclear rim of cells infected with US3 null and kinase-dead mutants. HSV-1 infection induced the phosphorylation of both lamin B and PKC. Elevated lamin B phosphorylation in HSV-1-infected cells was partially reduced by inhibitors of PKC. The data suggest a model in which kinases that normally disassemble the nuclear lamina during apoptosis are recruited to the nuclear membrane through functions requiring UL31 and UL34. We hypothesize that the recruitment of PKC functions to phosphorylate lamin B to help modify the nuclear lamina and promote budding of nucleocapsids at the inner nuclear membrane. PMID- 16352574 TI - The herpes simplex virus type 1 vhs-UL41 gene secures viral replication by temporarily evading apoptotic cellular response to infection: Vhs-UL41 activity might require interactions with elements of cellular mRNA degradation machinery. AB - We have previously shown that herpes simplex virus type 1 (HSV-1) infection is associated with early destabilization/degradation of infected cell mRNAs and consequent shutoff of host protein synthesis by the activity of the virion associated host shutoff (vhs) UL41 protein. Wild-type (wt) virus destabilized/degraded the housekeeping beta-actin and alpha-tubulin mRNAs as well host stress functions, like the heat shock 70 protein induced postinfection. vhs mutants did not degrade the mRNAs. Elaborate studies by others have been concerned with the mode of mRNA degradation and the mRNAs affected. We now describe vhs activity in primary cultures of mouse cerebellar granule neurons (CGNs). Specifically, (i) upon infection in the presence of actinomycin D to test activity of input viral particles, there was a generalized inhibition of protein synthesis, which depended on the input multiplicity of infection (MOI). (ii) Low MOI infection with vhs-1 mutant virus was associated with increased synthesis of all apparent proteins. Higher MOIs caused some shutoff, albeit significantly lower than that of wt virus. This pattern could reflect an interaction(s) of vhs 1 protein with host machinery involved in cellular mRNA destabilization/degradation, sequestering this activity. (iii) wt virus infection was associated with cell survival, at least for a while, whereas mutant virus induced apoptotic cell death at earlier times. (iv) wt virus replicated well in the CGNs, whereas there was no apparent replication of the vhs-1 mutant virus. (v) The vhs-1 mutant could serve as helper virus for composite amplicon vectors carrying marker genes and the human p53 gene. Ongoing studies test the use of vhs 1-based composite oncolytic vectors towards cancer gene therapy. PMID- 16352575 TI - Crimean-congo hemorrhagic fever virus glycoprotein precursor is cleaved by Furin like and SKI-1 proteases to generate a novel 38-kilodalton glycoprotein. AB - Crimean-Congo hemorrhagic fever virus (genus Nairovirus, family Bunyaviridae) genome M segment encodes an unusually large (in comparison to members of other genera) polyprotein (1,684 amino acids in length) containing the two major structural glycoproteins, Gn and Gc, that are posttranslationally processed from precursors PreGn and PreGc by SKI-1 and SKI-1-like proteases, respectively. The characteristics of the N-terminal 519 amino acids located upstream of the mature Gn are unknown. A highly conserved furin/proprotein convertase (PC) cleavage site motif (RSKR247) is located between the variable N-terminal region that is predicted to have mucin-like properties and the rest of PreGn. Mutational analysis of the RSKR247 motif and use of a specific furin/PC inhibitor and brefeldin A demonstrate that furin/PC cleavage occurs at the RSKR247 motif of PreGn as the protein transits the trans Golgi network and generates a novel glycoprotein designated GP38. Immunoprecipitation analysis identified two additional proteins, GP85 and GP160, which contain both mucin and GP38 domain regions, and whose generation does not involve furin/PC cleavage. Consistent with glycosylation predictions, heavy O-linked glycosylation and moderate levels of N glycans were detected in the GP85 and GP160 proteins, both of which contain the mucin domain. GP38, GP85, and GP160 are likely soluble proteins based on the lack of predicted transmembrane domains, their detection in virus-infected cell supernatants, and the apparent absence from virions. Analogy with soluble glycoproteins and mucin-like proteins encoded by other hemorrhagic fever associated RNA viruses suggests these proteins could play an important role in viral pathogenesis. PMID- 16352576 TI - Simian immunodeficiency virus Vpx is imported into the nucleus via importin alpha dependent and -independent pathways. AB - Vpx protein of human immunodeficiency virus type 2/simian immunodeficiency virus (SIV) has been implicated in the transport of the viral genome into the nuclei of nondividing cells. The mechanism by which Vpx enters the nucleus remains unknown. Here we have identified two distinct noncanonical nuclear localization signals (NLSs) in Vpx of SIV(smPbj1.9) and defined the pathways for its nuclear import. Although nuclear targeting signals identified here are distinct from known nuclear import signals, translocation of Vpx into the nucleus involves the interaction of its N-terminal NLS (amino acids 20 to 40) or C-terminal NLS (amino acids 65 to 75) with importin alpha and, in the latter case, also with importin beta. Collectively, these results suggest that importins interact with Vpx and ensure the effective import of Vpx into the nucleus to support virus replication in nondividing cells. PMID- 16352577 TI - Respiratory syncytial virus (RSV) infects neuronal cells and processes that innervate the lung by a process involving RSV G protein. AB - Respiratory syncytial virus (RSV) is a primary cause of morbidity and life threatening lower respiratory tract disease in infants and young children. Children with acute RSV bronchiolitis often develop respiratory sequelae, but the disease mechanisms are poorly understood. Mounting evidence suggests that RSV may mediate persistent infection. Using immunohistochemistry to identify RSV and RSV infected cell types, we show that RSV infects primary neurons and neuronal processes that innervate the lungs through a process that involves RSV G protein and the G protein CX3C motif. These findings suggest a mechanism for disease chronicity and have important implications for RSV disease intervention strategies. PMID- 16352578 TI - Gamma interferon primes productive human immunodeficiency virus infection in astrocytes. AB - Considerable controversy exists over whether astrocytes can support human immunodeficiency virus (HIV) infection. We evaluated the impact of three cytokines critical to the development of HIV neuropathogenesis, gamma interferon (IFN-gamma), granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor alpha, on priming astrocytes for HIV infection. We demonstrate that IFN gamma was the most potent in its ability to facilitate substantial productive HIV infection of an astroglioma cell line (U87MG) and human fetal astrocytes (HFA). The mechanism of IFN-gamma-mediated priming of HIV in HFA is unlikely to be at the level of up-regulation of receptors and coreceptors relevant to HIV entry. These data demonstrate that cytokine priming can alter HIV replication in astrocytes. PMID- 16352579 TI - Expression of m157, a murine cytomegalovirus-encoded putative major histocompatibility class I (MHC-I)-like protein, is independent of viral regulation of host MHC-I. AB - A murine cytomegalovirus (MCMV)-encoded protein, m157, has a putative major histocompatibility complex class I (MHC-I) structure and is recognized by the Ly49H NK cell activation receptor. Using a monoclonal antibody against m157, in this study we directly demonstrated that m157 is a cell surface-expressed glycophosphatidylinositol-anchored protein with early viral gene kinetics. Beta-2 microglobulin and TAP1 (transporter associated with antigen processing 1) were not required for its expression. MCMV-encoded proteins that down-regulate MHC-I did not affect the expression of m157. Thus, m157 is expressed on infected cells in a manner independent of viral regulation of host MHC-I. PMID- 16352580 TI - Quantification and reduction of cardiac dose in radical radiotherapy for oesophageal cancer. AB - Chemoradiation is increasingly used in the management of localized oesophageal cancer and has been shown in randomized controlled trials to improve overall survival. Although early toxicity of radiotherapy is well documented, this is not the case for late toxicity. As patients with oesophageal cancer have a high incidence of co-morbidities including cardiac problems, the aim of this paper was to quantify the extent of cardiac radiation and discuss the influence of beam arrangement to reduce this. Eight patients with localized oesophageal cancer treated with radical chemoradiation were selected. The mean cardiac dose and the volumes of heart receiving 30 Gy, 40 Gy and 45 Gy from the conventional two-phase technique were compared with those of single-phase 3-field and 4-field conformal beam arrangements. The 4-field arrangement reduced the mean cardiac dose by at least 3.3 Gy compared with the other two beam arrangements (p=0.01). The mean volume of heart receiving high doses between the three techniques widened as the dose increased in the range 30-45 Gy. There is no statistically significant difference in volumes receiving more than 30 Gy and 40 Gy. 65% of the cardiac volume received more than 45 Gy using a two-phase technique, compared with 57% using three fields and 26% using four fields (p<0.01). With a 4-field beam arrangement, therefore, there is a significant reduction in cardiac dose compared with the other two techniques. Cardiac toxicity and a 4-field beam arrangement should be considered when planning radical radiotherapy for localized oesophageal cancer. PMID- 16352581 TI - Parametrial invasion in carcinoma of cervix: role of MRI measured tumour volume. AB - The aim of the study was to determine the correlation between MRI measured tumour volume and parametrial invasion on histology in the evaluation of carcinoma of the cervix showing full thickness stromal invasion (FTSI). Original MR images of 159 surgical cases of carcinoma of the cervix retrieved from the MR image bank of the department were analysed retrospectively. Tumour volume from the maximum tumour diameters in each case was computed using the standard formula for an ellipse and correlated with parametrial invasion on histology. Of 105 cases showing FTSI on axial T(2) weighted MRI, tumour volume between the two groups of cases, with parametrial invasion (n=27) and without parametrial invasion (n=78), shows a statistically significant difference (p=0.006). Best accuracy of 60.95%, sensitivity 59.26% and specificity 61.54% was found for tumour volume of 36.39 cm(3). The study concludes that MRI measured tumour volume is associated with low accuracy in the evaluation of parametrial invasion in carcinoma of cervix showing FTSI in axial T(2) weighted MR images and may not help as an additional diagnostic criterion to predict parametrial invasion pre-operatively. PMID- 16352582 TI - Volume rendering of three-dimensional drip infusion CT cholangiography in patients with suspected obstructive biliary disease: a retrospective study. AB - The purpose of this study was to evaluate the diagnostic potential of prolonged drip infusion CT cholangiography (DIC-CT) using meglumine iotroxate (Biliscopin) and 3D volume rendering in patients with suspected obstructive biliary disease. From a material of 142 patients who had undergone a drip infusion CT, all cases with a verified surgical or endoscopic retrograde cholangiography (ERC) diagnosis (n=33) were selected. Age-matched controls were selected from the remaining examinations. Three radiologists reviewed all 66 examinations in retrospect, independently as well as in consensus. The image quality and the estimated diagnostic quality were rated as good or moderate in 91% of the 198 reviews. The consensus sensitivity and specificity for diagnosing biliary stones was 88% and 94%, respectively (with sensitivities ranging from 88% to 94% for individual observers, and specificities from 86% to 96%). Two out of three strictures were observed. No false positive strictures were described. The use of volume rendering technique (VRT) improved diagnostic certainty in 28/198 (14%) of the evaluations. The visualization of ductal stones was improved in 18/48 (38%). No differences in diagnostic quality between single and multislice CT were observed. We conclude that a detailed image of the biliary tree with good sensitivity and specificity can be obtained by means of bilirubin-governed infusion time DIC-CT with volume rendering reconstruction. PMID- 16352583 TI - Spatial mapping of the brachial plexus using three-dimensional ultrasound. AB - Imaging of the brachial plexus with MRI and standard two-dimensional (2D) ultrasound has been reported, and 2D ultrasound-guided regional anaesthetic block is an established technique. The aim of this study was to map the orientation of the brachial plexus in relation to the first rib, carotid and subclavian arteries, using three-dimensional (3D) ultrasound. A free-hand optically tracked 3D ultrasound system was used with a 12 MHz transducer. 10 healthy volunteers underwent 3D ultrasound of the neck. From the 3D ultrasound data sets, the outlines of the brachial plexus, subclavian artery and first rib were manually segmented. A surface was interpolated from the series of outlines to produce a spatially orientated 3D reconstruction of the brachial plexus. The brachial plexus could be mapped in all volunteers, although a variation in image resolution between individuals existed. Anatomical variations were demonstrated between the 10 volunteers; the most notable and clinically relevant was the alignment of the plexus divisions. 3D reconstructions illustrated the plexus, changing its orientation from a vertical alignment in the interscalene region to a more horizontal alignment in the supraclavicular fossa. Spatial mapping of the brachial plexus is possible with 3D ultrasound using the subclavian artery and first rib as landmarks. There is a deviation from the conventionally described anatomy and this may have implications for the administration of regional anaesthesia. PMID- 16352584 TI - Plexiform neurofibroma of the oesophagus: a mimicker of malignancy. AB - We report the first case of a plexiform neurofibroma of the oesophagus, presenting with dysphagia in a 67-year-old man known to suffer from von Recklinghausen's neurofibromatosis. The clinical symptoms and radiological findings mimicked malignancy, and raised considerable concern. Numerous investigations failed to elucidate the benign nature of the condition. The diagnosis was finally achieved at surgery and the patient was successfully treated by oesophageal resection. Plexiform neurofibromas, though only found in 20-30% of affected individuals are pathognomonic of von Recklinghausen's neurofibromatosis. This report highlights the tendency of plexiform neurofibromas to grow extensively and encase surrounding structures, thereby mimicking a neoplastic process. PMID- 16352585 TI - A pictorial review of the varied appearance of atypical liver metastasis from carcinoma of the breast. PMID- 16352586 TI - Magnetic resonance imaging appearances of fibrous dysplasia. AB - Fibrous dysplasia is a developmental disorder in which normal bone marrow is replaced by fibro-osseous tissue. The radiographic, CT and scintigraphic appearances of this condition are well known. The MRI appearances of fibrous dysplasia have not been widely published. The lesions are largely isointense with areas of hypointensity on T(1) weighted images and appear heterogeneously hyperintense on T(2) weighted images. The enhancement pattern is patchy central, rim, homogeneous, or a combination. The MRI features reflect the variable tissue components of this entity. This pictorial review aims at highlighting the MRI appearances, with pathological correlation. PMID- 16352587 TI - Multiple hepatosplenic nodules. PMID- 16352589 TI - Hepatic perfusion index measured using MRI and scintigraphy. PMID- 16352590 TI - Conventional wisdom and activities of the middle range. PMID- 16352593 TI - Targeting of Miz-1 is essential for Myc-mediated apoptosis. AB - The c-Myc oncoprotein plays a central role in human cancer via its ability to either activate or repress the transcription of essential downstream targets. For many of the repressed target genes, down-regulation by c-Myc relies on its ability to bind and inactivate the transcription factor Miz-1. Although Miz-1 inactivation is suspected to be essential for at least some of the biological activities of c-Myc, it has been difficult to demonstrate this requirement experimentally. Using a combination of short hairpin RNA-mediated knockdown and a previously characterized mutant of c-Myc that is defective for Miz-1 inactivation, we examined whether this inactivation is critical for three of the most central biological functions of c-Myc, cell cycle progression, transformation, and apoptosis. The results of this analysis demonstrated that in the in vitro assays utilized here, Miz-1 inactivation is dispensable for c-Myc induced cell cycle progression and transformation. In marked contrast, the ability of c-Myc to induce apoptosis in primary diploid human fibroblasts in response to growth factor withdrawal is entirely dependent on its ability to inactivate Miz-1. These data have a significant impact on our understanding of the biochemical mechanisms dictating how c-Myc mediates opposing biological functions, such as transformation and apoptosis, and demonstrate the first requirement for Miz-1 inactivation in any of the biological functions of c-Myc. PMID- 16352594 TI - Ezrin directly interacts with the alpha1b-adrenergic receptor and plays a role in receptor recycling. AB - Using the yeast two-hybrid system, we identified ezrin as a protein interacting with the C-tail of the alpha1b-adrenergic receptor (AR). The interaction was shown to occur in vitro between the receptor C-tail and the N-terminal portion of ezrin, or Four-point-one ERM (FERM) domain. The alpha1b-AR/ezrin interaction occurred inside the cells as shown by the finding that the transfected alpha1b-AR and FERM domain or ezrin could be coimmunoprecipitated from human embryonic kidney 293 cell extracts. Mutational analysis of the alpha1b-AR revealed that the binding site for ezrin involves a stretch of at least four arginines on the receptor C-tail. The results from both receptor biotinylation and immunofluorescence experiments indicated that the FERM domain impaired alpha1b-AR recycling to the plasma membrane without affecting receptor internalization. The dominant negative effect of the FERM domain, which relies on its ability to mask the ezrin binding site for actin, was mimicked by treatment of cells with cytochalasin D, an actin depolymerizing agent. A receptor mutant (DeltaR8) lacking its binding site in the C-tail for ezrin displayed delayed receptor recycling. These findings identify ezrin as a new protein directly interacting with a G protein-coupled receptor and demonstrate the direct implication of ezrin in GPCR trafficking via an actin-dependent mechanism. PMID- 16352595 TI - Dual roles for the phosphatase PPM1D in regulating progesterone receptor function. AB - Although protein phosphatase magnesium-dependent 1 delta (PPM1D) was initially characterized as a p53-regulated phosphatase responsible for inactivation of p38 MAPK and consequent inactivation of p53, its overexpression and amplification in human breast cancers led us to assess its role in steroid hormone action. We found that PPM1D stimulated the activity of several nuclear receptors including the progesterone receptor (PR) and estrogen receptor. Although p38 MAPK inhibited PR activity, PPM1D stimulation of PR activity was greater than that achieved by a chemical inhibitor of p38 MAPK, SB202190. This suggests an additional novel function for PPM1D. Consistent with this, the transcriptional activity of endogenous PR in MCF-7 breast cancer cells was preferentially inhibited by small interfering RNA for PPM1D; SB202190 failed to reverse the inhibition. Although PPM1D phosphatase activity was required for stimulation of transcriptional activity, the activity of a PR phosphorylation site null mutant was enhanced by PPM1D, indicating that PR is not the direct target. Additional studies revealed that PPM1D enhanced the intrinsic activity of p160 coactivators such as steroid receptor coactivator-1 and promoted the interaction between PR and steroid receptor coactivator-1 in a mammalian two-hybrid assay. Neither activity was induced by SB202190. Although PPM1D stimulated PR activity in part through inhibition of p38 MAPK, its primary action is novel and independent of p38 MAPK. Thus, we speculate that PPM1D promotes breast tumor growth both by inhibiting p53 activity and by enhancing steroid hormone receptor action. PMID- 16352596 TI - Sodium-induced GCN4 expression controls the accumulation of the 5' to 3' RNA degradation inhibitor, 3'-phosphoadenosine 5'-phosphate. AB - Most cytoplasmic mRNAs are decapped and digested by the 5'-3'-exonuclease Xrn1p in Saccharomyces cerevisiae. The activity of Xrn1p is naturally inhibited in the presence of 3'-phosphoadenosine 5'-phosphate (pAp), a metabolite produced during sulfate assimilation that is quickly metabolized to AMP by the enzymatic activity of Hal2p. However, pAp accumulates and 5'-3' degradation decreases in the presence of ions known to inhibit Hal2p activity, such as sodium or lithium. We have shown that yeast cells can better adapt to the presence of sodium than lithium because of their ability to reduce pAp accumulation by activating HAL2 expression in a Gcn4p-dependent response, a regulatory loop that is likely to be conserved in different yeast species. We have thus identified a new role for the transcriptional activity of Gcn4p in maintaining an active mRNA degradation pathway under conditions of sodium stress. Since deregulation of proteins involved in different metabolic pathways is observed in xrn1Delta mutants, the maintenance of mRNA degradation capacity is likely to be important for the accurate and rapid adaptation of gene expression to salt stress. PMID- 16352597 TI - Crystal structure of human cytochrome P450 2D6. AB - Cytochrome P450 2D6 is a heme-containing enzyme that is responsible for the metabolism of at least 20% of known drugs. Substrates of 2D6 typically contain a basic nitrogen and a planar aromatic ring. The crystal structure of human 2D6 has been solved and refined to 3.0A resolution. The structure shows the characteristic P450 fold as seen in other members of the family, with the lengths and orientations of the individual secondary structural elements being very similar to those seen in 2C9. There are, however, several important differences, the most notable involving the F helix, the F-G loop, the B'helix, beta sheet 4, and part of beta sheet 1, all of which are situated on the distal face of the protein. The 2D6 structure has a well defined active site cavity above the heme group, containing many important residues that have been implicated in substrate recognition and binding, including Asp-301, Glu-216, Phe-483, and Phe-120. The crystal structure helps to explain how Asp-301, Glu-216, and Phe-483 can act as substrate binding residues and suggests that the role of Phe-120 is to control the orientation of the aromatic ring found in most substrates with respect to the heme. The structure has been compared with published homology models and has been used to explain much of the reported site-directed mutagenesis data and help understand the metabolism of several compounds. PMID- 16352598 TI - Involvement of the mouse Prp19 gene in neuronal/astroglial cell fate decisions. AB - The molecular mechanisms involved in neuronal/astroglial cell fate decisions during the development of the mammalian central nervous system are poorly understood. Here, we report that PRP19beta, a splice variant of mouse PRP19alpha corresponding to the yeast PRP19 protein, can function as a neuron-astroglial switch during the retinoic acid-primed neural differentiation of P19 cells. The beta-variant possesses an additional 19 amino acid residues in-frame in the N terminal region of the alpha-variant. The forced expression of the alpha-variant RNA caused the down-regulation of oct-3/4 and nanog mRNA expression during the 12 48 h of the late-early stages of neural differentiation and was sufficient to convert P19 cells into neurons (but not glial cells) when the cells were cultured in aggregated form without retinoic acid. In contrast, the forced expression of the beta-variant RNA suppressed neuronal differentiation and conversely stimulated astroglial cell differentiation in retinoic acid-primed P19 cells. Based on yeast two-hybrid screening, cyclophilin A was identified as a specific binding partner of the beta-variant. Luciferase reporter assay mediated by the oct-3/4 promoter revealed that cyclophilin A could act as a transcriptional activator and that its activity was suppressed by the beta-variant, suggesting that cyclophilin A takes part in the induction of oct-3/4 gene expression, which might lead to neuroectodermal otx2 expression within 12 h of the immediate-early stages of retinoic acid-primed neural differentiation. These results show that the alpha-variant gene plays a pivotal role in neural differentiation and that the beta-variant participates in neuronal/astroglial cell fate decisions. PMID- 16352599 TI - The interferon-inducible ubiquitin-protein isopeptide ligase (E3) EFP also functions as an ISG15 E3 ligase. AB - The expression of the ubiquitin-like protein ISG15 and protein modification by ISG15 (ISGylation) are strongly activated by interferons. Accordingly, ISG15 expression and protein ISGylation are strongly activated upon viral and bacterial infections and during other stress conditions, suggesting important roles for the ISG15 system in innate immune responses. Here, we report the identification of the ubiquitin-protein isopeptide ligase (E3) EFP (estrogen-responsive finger protein) as the ISG15 E3 ligase for 14-3-3sigma protein. Like other known components of the protein ISGylation system (ISG15, UBE1L, UBP43, and UBC8), EFP is also an interferon-inducible protein. Expression of EFP small interfering RNA decreased the ISGylation of 14-3-3sigma in the 293T cell ISGylation system as well as in MCF-7 cells upon interferon treatment. Furthermore, the ISGylation enzyme activity of EFP was RING domain-dependent. These findings indicate that EFP is an ISG15 E3 ligase for 14-3-3sigma in vivo. The fact that both UBC8 and EFP are common components in the ubiquitin and ISG15 conjugation pathways suggests a mechanism whereby a limited set of enzymes accomplishes diverse post translational modifications of their substrates in response to changes in environmental stimulations. PMID- 16352600 TI - The expanded octarepeat domain selectively binds prions and disrupts homomeric prion protein interactions. AB - Insertion of additional octarepeats into the prion protein gene has been genetically linked to familial Creutzfeldt Jakob disease and hence to de novo generation of infectious prions. The pivotal event during prion formation is the conversion of the normal prion protein (PrPC) into the pathogenic conformer PrPSc, which subsequently induces further conversion in an autocatalytic manner. Apparently, an expanded octarepeat domain directs folding of PrP toward the PrPSc conformation and initiates a self-replicating conversion process. Here, based on three main observations, we have provided a model on how altered molecular interactions between wild-type and mutant PrP set the stage for familial Creutzfeldt Jakob disease with octarepeat insertions. First, we showed that wild type octarepeat domains interact in a copper-dependent and reversible manner, a "copper switch." This interaction becomes irreversible upon domain expansion, possibly reflecting a loss of function. Second, expanded octarepeat domains of increasing length gradually form homogenous globular multimers of 11-21 nm in the absence of copper ions when expressed as soluble glutathione S-transferase fusion proteins. Third, octarepeat domain expansion causes a gain of function with at least 10 repeats selectively binding PrPSc in a denaturant-resistant complex in the absence of copper ions. Thus, the combination of both a loss and gain of function profoundly influences homomeric interaction behavior of PrP with an expanded octarepeat domain. A multimeric cluster of prion proteins carrying expanded octarepeat domains may therefore capture and incorporate spontaneously arising short-lived PrPSc-like conformers, thereby providing a matrix for their conversion. PMID- 16352601 TI - Mutations conferring resistance to SCH6, a novel hepatitis C virus NS3/4A protease inhibitor. Reduced RNA replication fitness and partial rescue by second site mutations. AB - Drug resistance is a major issue in the development and use of specific antiviral therapies. Here we report the isolation and characterization of hepatitis C virus RNA replicons resistant to a novel ketoamide inhibitor of the NS3/4A protease, SCH6 (originally SCH446211). Resistant replicon RNAs were generated by G418 selection in the presence of SCH6 in a dose-dependent fashion, with the emergence of resistance reduced at higher SCH6 concentrations. Sequencing demonstrated remarkable consistency in the mutations conferring SCH6 resistance in genotype 1b replicons derived from two different strains of hepatitis C virus, A156T/A156V and R109K. R109K, a novel mutation not reported previously to cause resistance to NS3/4A inhibitors, conferred moderate resistance only to SCH6. Structural analysis indicated that this reflects unique interactions of SCH6 with P'-side residues in the protease active site. In contrast, A156T conferred high level resistance to SCH6 and a related ketoamide, SCH503034, as well as BILN 2061 and VX-950. Unlike R109K, which had minimal impact on NS3/4A enzymatic function, A156T significantly reduced NS3/4A catalytic efficiency, polyprotein processing, and replicon fitness. However, three separate second-site mutations, P89L, Q86R, and G162R, were capable of partially reversing A156T-associated defects in polyprotein processing and/or replicon fitness, without significantly reducing resistance to the protease inhibitor. PMID- 16352602 TI - Defining the role of the Escherichia coli chaperone SecB using comparative proteomics. AB - To improve understanding and identify novel substrates of the cytoplasmic chaperone SecB in Escherichia coli, we analyzed a secB null mutant using comparative proteomics. The secB null mutation did not affect cell growth but caused significant differences at the proteome level. In the absence of SecB, dynamic protein aggregates containing predominantly secretory proteins accumulated in the cytoplasm. Unprocessed secretory proteins were detected in radiolabeled whole cell lysates. Furthermore, the assembly of a large fraction of the outer membrane proteome was slowed down, whereas its steady state composition was hardly affected. In response to aggregation and delayed sorting of secretory proteins, cytoplasmic chaperones DnaK, GroEL/ES, ClpB, IbpA/B, and HslU were up regulated severalfold, most likely to stabilize secretory proteins during their delayed translocation and/or rescue aggregated secretory proteins. The SecB/A dependence of 12 secretory proteins affected by the secB null mutation (DegP, FhuA, FkpA, OmpT, OmpX, OppA, TolB, TolC, YbgF, YcgK, YgiW, and YncE) was confirmed by "classical" pulse-labeling experiments. Our study more than triples the number of known SecB-dependent secretory proteins and shows that the primary role of SecB is to facilitate the targeting of secretory proteins to the Sec translocase. PMID- 16352603 TI - The nerve hemoglobin of the bivalve mollusc Spisula solidissima: molecular cloning, ligand binding studies, and phylogenetic analysis. AB - Members of the hemoglobin (Hb) superfamily are present in nerve tissue of several vertebrate and invertebrate species. In vertebrates they display hexacoordinate heme iron atoms and are typically expressed at low levels (microM). Their function is still a matter of debate. In invertebrates they have a hexa- or pentacoordinate heme iron, are mostly expressed at high levels (mM), and have been suggested to have a myoglobin-like function. The native Hb of the surf clam, Spisula solidissima, composed of 162 amino acids, does not show specific deviations from the globin templates. UV-visible and resonance Raman spectroscopy demonstrate a hexacoordinate heme iron. Based on the sequence analogy, the histidine E7 is proposed as a sixth ligand. Kinetic and equilibrium measurements show a moderate oxygen affinity (P(50) approximately 0.6 torr) and no cooperativity. The histidine binding affinity is 100-fold lower than in neuroglobin. Phylogenetic analysis demonstrates a clustering of the S. solidissima nerve Hb with mollusc Hbs and myoglobins, but not with the vertebrate neuroglobins. We conclude that invertebrate nerve Hbs expressed at high levels are, despite the hexacoordinate nature of their heme iron, not essentially different from other intracellular Hbs. They most likely fulfill a myoglobin-like function and enhance oxygen supply to the neurons. PMID- 16352604 TI - Expression of ABCG1, but not ABCA1, correlates with cholesterol release by cerebellar astroglia. AB - Central nervous system lipoproteins mediate the exchange of cholesterol between cells and support synaptogenesis and neuronal growth. The primary source of lipoproteins in the brain is astroglia cells that synthesize and secrete apolipoprotein (apo) E in high density lipoprotein-like particles. Small quantities of apoA1, derived from the peripheral circulation, are also present in the brain. In addition to the direct secretion of apoE-containing lipoproteins from astroglia, glia-derived lipoproteins are thought to be formed by cholesterol efflux to extracellular apolipoproteins via ATP-binding cassette (ABC) transporters. We used cultured cerebellar murine astroglia to investigate the relationship among cholesterol availability, apoE secretion, expression of ABCA1 and ABCG1, and cholesterol efflux. In many cell types, cholesterol content, ABCA1 expression, and cholesterol efflux are closely correlated. In contrast, cholesterol enrichment of glia failed to increase ABCA1 expression, although ABCG1 expression and cholesterol efflux to apoA1 were increased. Moreover, the liver X receptor (LXR) agonist TO901317 up-regulated ABCA1 and ABCG1 expression in glia without stimulating cholesterol efflux. Larger lipoproteins were generated when glia were enriched with cholesterol, whereas treatment with the LXR agonist produced smaller particles that were eliminated when the glia were loaded with cholesterol. We also used glia from ApoE(-/-) mice to distinguish between direct lipoprotein secretion and the extracellular generation of lipoproteins. Our observations indicate that partially lipidated apoE, secreted directly by glia, is likely to be the major extracellular acceptor of cholesterol released from glia in a process mediated by ABCG1. PMID- 16352605 TI - Rapid loss of structural motifs in the manganese complex of oxygenic photosynthesis by X-ray irradiation at 10-300 K. AB - Structural changes upon photoreduction caused by x-ray irradiation of the water oxidizing tetramanganese complex of photosystem II were investigated by x-ray absorption spectroscopy at the manganese K-edge. Photoreduction was directly proportional to the x-ray dose. It was faster in the higher oxidized S2 state than in S1; seemingly the oxidizing potential of the metal site governs the rate. X-ray irradiation of the S1 state at 15 K initially caused single-electron reduction to S0* accompanied by the conversion of one di-mu-oxo bridge between manganese atoms, previously separated by approximately 2.7 A, to a mono-mu-oxo motif. Thereafter, manganese photoreduction was 100 times slower, and the biphasic increase in its rate between 10 and 300 K with a breakpoint at approximately 200 K suggests that protein dynamics is rate-limiting the radical chemistry. For photoreduction at similar x-ray doses as applied in protein crystallography, halfway to the final Mn(II)4 state the complete loss of inter manganese distances <3 A was observed, even at 10 K, because of the destruction of mu-oxo bridges between manganese ions. These results put into question some structural attributions from recent protein crystallography data on photosystem II. It is proposed to employ controlled x-ray photoreduction in metalloprotein research for: (i) population of distinct reduced states, (ii) estimating the redox potential of buried metal centers, and (iii) research on protein dynamics. PMID- 16352606 TI - Caspase-7 is directly activated by the approximately 700-kDa apoptosome complex and is released as a stable XIAP-caspase-7 approximately 200-kDa complex. AB - MCF-7 cells lack caspase-3 but undergo mitochondrial-dependent apoptosis via caspase-7 activation. It is assumed that the Apaf-1-caspase-9 apoptosome processes caspase-7 in an analogous manner to that described for caspase-3. However, this has not been validated experimentally, and we have now characterized the caspase-7 activating apoptosome complex in MCF-7 cell lysates activated with dATP/cytochrome c. Apaf-1 oligomerizes to produce approximately 1.4-MDa and approximately 700-kDa apoptosome complexes, and the latter complex directly cleaves/activates procaspase-7. This approximately 700-kDa apoptosome complex, which is also formed in apoptotic MCF-7 cells, is assembled by rapid oligomerization of Apaf-1 and followed by a slower process of procaspase-9 recruitment and cleavage to form the p35/34 forms. However, procaspase-9 recruitment and processing are accelerated in lysates supplemented with caspase 3. In lysates containing very low levels of Smac and Omi/HtrA2, XIAP (X-linked inhibitor of apoptosis) binds tightly to caspase-9 in the apoptosome complex, and as a result caspase-7 processing is abrogated. In contrast, in MCF-7 lysates containing Smac and Omi/HtrA2, active caspase-7 is released from the apoptosome and forms a stable approximately 200-kDa XIAP-caspase-7 complex, which apparently does not contain cIAP1 or cIAP2. Thus, in comparison to caspase-3-containing cells, XIAP appears to have a more significant antiapoptotic role in MCF-7 cells because it directly inhibits caspase-7 activation by the apoptosome and also forms a stable approximately 200-kDa complex with active caspase-7. PMID- 16352607 TI - Functional asymmetry of nucleotide-binding domains in ABCG5 and ABCG8. AB - The ATP-binding cassette half-transporters ABCG5 (G5) and ABCG8 (G8) promote secretion of neutral sterols into bile, a major pathway for elimination of sterols. Mutations in either ABCG5 or ABCG8 cause sitosterolemia, a recessive disorder characterized by impaired biliary and intestinal sterol secretion, sterol accumulation, and premature atherosclerosis. The mechanism by which the G5G8 heterodimer couples ATP hydrolysis to sterol transport is not known. Here we examined the roles of the Walker A, Walker B, and signature motifs in the nucleotide-binding domains (NBD) of G5 and G8 using recombinant adenoviruses to reconstitute biliary sterol transport in G5G8-deficient mice. Mutant forms of each half-transporter were co-expressed with their wild-type partners. Mutations at crucial residues in the Walker A and Walker B domains of G5 prevented biliary sterol secretion, whereas mutations of the corresponding residues in G8 did not. The opposite result was obtained when mutations were introduced into the signature motif; mutations in the signature domain of G8 prevented sterol transport, but substitution of the corresponding residues in G5 did not. Taken together, these findings indicate that the NBDs of G5 and G8 are not functionally equivalent. The integrity of the canonical NBD formed by the Walker A and Walker B motifs of G5 and the signature motif of G8 is essential for G5G8-mediated sterol transport. In contrast, mutations in key residues of the NBD formed by the Walker A and B motifs of G8 and the signature sequence of G5 did not affect sterol secretion. PMID- 16352608 TI - ATP induces conformational changes of periplasmic loop regions of the maltose ATP binding cassette transporter. AB - We have studied cofactor-induced conformational changes of the maltose ATP binding cassette transporter by employing limited proteolysis in detergent solution. The transport complex consists of one copy each of the transmembrane subunits, MalF and MalG, and of two copies of the nucleotide-binding subunit, MalK. Transport activity further requires the periplasmic maltose-binding protein, MalE. Binding of ATP to the MalK subunits increased the susceptibility of two tryptic cleavage sites in the periplasmic loops P2 of MalF and P1 of MalG, respectively. Lys(262) of MalF and Arg(73) of MalG were identified as probable cleavage sites, resulting in two N-terminal peptide fragments of 29 and 8 kDa, respectively. Trapping the complex in the transition state by vanadate further stabilized the fragments. In contrast, the tryptic cleavage profile of MalK remained largely unchanged. ATP-induced conformational changes of MalF-P2 and MalG-P1 were supported by fluorescence spectroscopy of complex variants labeled with 2-(4'-maleimidoanilino)naphthalene-6-sulfonic acid. Limited proteolysis was subsequently used as a tool to study the consequences of mutations on the transport cycle. The results suggest that complex variants exhibiting a binding protein-independent phenotype (MalF500) or containing a mutation that affects the "catalytic carboxylate" (MalKE159Q) reside in a transition state-like conformation. A similar conclusion was drawn for a complex containing a replacement of MalKQ140 in the signature sequence by leucine, whereas substitution of lysine for Gln(140) appears to lock the transport complex in the ground state. Together, our data provide the first evidence for conformational changes of the transmembrane subunits of an ATP-binding cassette import system upon binding of ATP. PMID- 16352609 TI - Kinetics of the ATP hydrolysis cycle of the nucleotide-binding domain of Mdl1 studied by a novel site-specific labeling technique. AB - We have recently proposed a "processive clamp" model for the ATP hydrolysis cycle of the nucleotide-binding domain (NBD) of the mitochondrial ABC transporter Mdl1 (Janas, E., Hofacker, M., Chen, M., Gompf, S., van der Does, C., and Tampe, R. (2003) J. Biol. Chem. 278, 26862-26869). In this model, ATP binding to two monomeric NBDs leads to formation of an NBD dimer that, after hydrolysis of both ATPs, dissociates and releases ADP. Here, we set out to follow the association and dissociation of NBDs using a novel minimally invasive site-specific labeling technique, which provides stable and stoichiometric attachment of fluorophores. The association and dissociation kinetics of the E599Q-NBD dimer upon addition and removal of ATP were determined by fluorescence self-quenching. Remarkably, the rate of ATP hydrolysis of the wild type NBD is determined by the rate of NBD dimerization. In the E599QNBD, however, in which the ATP hydrolysis is 250-fold reduced, the ATP hydrolysis reaction controls dimer dissociation and the overall ATPase cycle. These data explain contradicting observations on the rate-limiting step of various ABC proteins and further demonstrate that dimer formation is an important step in the ATP hydrolysis cycle. PMID- 16352610 TI - Topology of the porin MspA in the outer membrane of Mycobacterium smegmatis. AB - MspA is the major porin of Mycobacterium smegmatis mediating the exchange of hydrophilic solutes across the outer membrane (OM). It is the prototype of a new family of octameric porins with a single central channel of 9.6 nm in length and consists of two hydrophobic beta-barrels of 3.7 nm in length and a more hydrophilic, globular rim domain. The length of the hydrophobic domain of MspA does not match the thicknesses of mycobacterial OMs of 5-12 nm as derived from electron micrographs. Further, the membrane topology of MspA is unknown as it is for any other mycobacterial OM protein. We used MspA as a molecular ruler to define the boundaries of the OM of M. smegmatis by surface labeling of single cysteine mutants. Seventeen mutants covered the surface of the rim domain and were biotinylated with a membrane-impermeable reagent. The label efficiencies in vitro were remarkably similar to the predicted accessibilities of the cysteines. By contrast, six of these mutants were protected from biotinylation in M. smegmatis cells. Tryptophan 21 defines a horizontal plane that dissects the surface-exposed versus the membrane-protected residues of MspA. The 8 phenylalanines at position 99 form a ring at the periplasmic end of the hydrophobic beta-barrel domain. These results indicated that (i) the membrane boundaries of MspA are defined by aromatic girdles as in porins of Gram-negative bacteria and (ii) loops and a 3.4-nm long part of the hydrophilic rim domain are embedded into the OM of M. smegmatis. This is the first report suggesting that elements other than hydrophobic alpha-helices or beta-sheets are integrated into a lipid membrane. PMID- 16352611 TI - Hepatocyte growth factor-regulated tyrosine kinase substrate (HRS) interacts with PELP1 and activates MAPK. AB - PELP1 (proline-, glutamic acid-, and leucine-rich protein-1) (also known as the modulator of nongenomic activity of estrogen receptor) plays a role in genomic functions of the estrogen receptor via histone interactions and in nongenomic functions via its influence on the MAPK-Src pathway. However, recent studies have shown that differential compartmentalization of PELP1 could play a crucial role in modulating the status of nongenomic signaling by using molecular mechanisms that remain poorly understood. Hepatocyte growth factor-regulated tyrosine kinase substrate (HRS) is an early endosomal protein that plays a role in regulating the trafficking of growth factor-receptor complexes through early endosomes. By using a yeast two-hybrid screen, we identified HRS as a novel PELP1-binding protein providing evidence of a physiologic interaction between HRS and PELP1. The noted HRS-PELP1 interaction was accompanied by inhibition of the basal coactivator function of PELP1 upon estrogen receptor transactivation. HRS was found to sequester PELP1 in the cytoplasm, leading to the activation of MAPK in a manner that is dependent on the epidermal growth factor receptor but independent of the estrogen receptor, Shc, and Src. In addition, stimulation of MAPK and the subsequent activation of its downstream effector pathway, Elk-1, by HRS or PELP1 were found to depend on the presence of endogenous PELP1 or HRS. Furthermore, HRS was overexpressed and correlated well with the cytoplasmic PELP1, increased MAPK, and EGFR status in breast tumors. These findings highlight a novel role of HRS in up-regulating MAPK, presumably involving interaction with PELP1. PMID- 16352612 TI - Stochastic high-speed rotation of Escherichia coli ATP synthase F1 sector: the epsilon subunit-sensitive rotation. AB - The gamma subunit of the ATP synthase F(1) sector rotates at the center of the alpha(3)beta(3) hexamer during ATP hydrolysis. A gold bead (40-200 nm diameter) was attached to the gamma subunit of Escherichia coli F(1), and then its ATP hydrolysis-dependent rotation was studied. The rotation speeds were variable, showing stochastic fluctuation. The high-speed rates of 40- and 60-nm beads were essentially similar: 721 and 671 rps (revolutions/s), respectively. The average rate of 60-nm beads was 381 rps, which is approximately 13-fold faster than that expected from the steady-state ATPase turnover number. These results indicate that the F(1) sector rotates much faster than expected from the bulk of ATPase activity, and that approximately 10% of the F(1) molecules are active on the millisecond time scale. Furthermore, the real ATP turnover number (number of ATP molecules converted to ADP and phosphate/s), as a single molecule, is variable during a short period. The epsilon subunit inhibited rotation and ATPase, whereas epsilon fused through its carboxyl terminus to cytochrome b(562) showed no effect. The epsilon subunit significantly increased the pausing time during rotation. Stochastic fluctuation of catalysis may be a general property of an enzyme, although its understanding requires combining studies of steady-state kinetics and single molecule observation. PMID- 16352613 TI - Divergent mechanisms utilized by SOCS3 to mediate interleukin-10 inhibition of tumor necrosis factor alpha and nitric oxide production by macrophages. AB - The cytokine interleukin-10 (IL-10) potently inhibits macrophage function through activation of the transcription factor STAT3. The expression of SOCS3 (suppressor of cytokine signaling-3) has been shown to be induced by IL-10 in a STAT3 dependent manner. However, the relevance of SOCS3 expression to the anti inflammatory effect of IL-10 on macrophages has been controversial. Through kinetic analysis of the requirement for SOCS3 in IL-10 inhibition of lipopolysaccharide (LPS)-stimulated tumor necrosis factor-alpha (TNFalpha) transcription and translation, SOCS3 was found to be necessary for TNFalpha expression during the early phase, but not the late phase of IL-10 action. SOCS3 was essential for IL-10 inhibition of LPS-stimulated production of iNOS (inducible nitric-oxide synthase) protein and nitric oxide (NO). To determine the domains of SOCS3 protein important in mediating these effects, SOCS3-/- macrophages were reconstituted with SOCS3 mutated for the SH2, KIR, SOCS box domains, and tyrosines 204 (Tyr204) and 221 (Tyr221). The SH2 domain, SOCS box, and both Tyr204 and Tyr221 were required for IL-10 inhibition of TNFalpha mRNA and protein expression, but interestingly the KIR domain was necessary only for IL-10 inhibition of TNFalpha protein expression. In contrast, Tyr204 and Tyr221 were the only structural features of SOCS3 that were necessary in mediating IL-10 inhibition of iNOS protein expression and NO production. These data define SOCS3 as an important mediator of IL-10 inhibition of macrophage activation and that SOCS3 interferes with distinct LPS-stimulated signal transduction events through differing mechanisms. PMID- 16352614 TI - Expression of FLR1 transporter requires phospholipase C and is repressed by Mediator. AB - In budding yeast, phosphoinositide-specific phospholipase C (Plc1p encoded by PLC1 gene) is important for function of kinetochores. Deletion of PLC1 results in benomyl sensitivity, alterations in chromatin structure of centromeres, mitotic delay, and a higher frequency of chromosome loss. Here we intended to utilize benomyl sensitivity as a phenotype that would allow us to identify genes that are important for kinetochore function and are downstream of Plc1p. However, our screen identified SIN4, encoding a component of the Mediator complex of RNA polymerase II. Deletion of SIN4 gene (sin4Delta) does not suppress benomyl sensitivity of plc1Delta cells by improving the function of kinetochores. Instead, benomyl sensitivity of plc1Delta cells is caused by a defect in expression of FLR1, and the suppression of benomyl sensitivity in plc1Delta sin4Delta cells occurs by derepression of FLR1 transcription. FLR1 encodes a plasma membrane transporter that mediates resistance to benomyl. Several other mutations in the Mediator complex also result in significant derepression of FLR1 and greatly increased resistance to benomyl. Thus, benomyl sensitivity is not a phenotype exclusively associated with mitotic spindle defect. These results demonstrate that in addition to promoter-specific transcription factors that are components of the pleiotropic drug resistance network, expression of the membrane transporters can be regulated by Plc1p, a component of a signal transduction pathway, and by Mediator, a general transcription factor. The results thus suggest another layer of complexity in regulation of pleiotropic drug resistance. PMID- 16352615 TI - Observation of a unique pattern of bifurcated hydrogen bonds in the crystal structures of the N-glycoprotein linkage region models. AB - Elucidation of the intra- and intermolecular carbohydrate-protein interactions would greatly contribute toward obtaining a better understanding of the structure function correlations of the protein-linked glycans. The weak interactions involving C-H...O have recently been attracting immense attention in the domain of biomolecular recognition. However, there has been no report so far on the occurrence of C-H...O hydrogen bonds in the crystal structures of models and analogs of N-glycoproteins. We present herein an analysis of C-H...O interactions in the crystal structures of all N-glycoprotein linkage region models and analogs. The study reveals a cooperative network of bifurcated hydrogen bonds consisting of N-H...O and C-H...O interactions seen uniquely for the models. The cooperative network consists of two antiparallel chains of bifurcated hydrogen bonds, one involving N1-H, C2'-H and O1' of the aglycon moiety and the other involving N2-H, C1-H and O1'' of the sugar. Such bifurcated hydrogen bonds between the core glycan and protein are likely to play an important role in the folding and stabilization of proteins. PMID- 16352616 TI - Molecular subtypes of bladder cancer: Jekyll and Hyde or chalk and cheese? AB - Cancer of the bladder shows divergent clinical behaviour following diagnosis and it has been proposed that two major groups of tumours exist that develop via different molecular pathways. Low-grade, non-invasive papillary tumours recur frequently, but patients with these tumours do not often suffer progression of disease to muscle invasion. In contrast, tumours that are invading muscle at diagnosis are aggressive and associated with significant mortality. Molecular studies have identified distinct genetic, epigenetic and expression changes in these groups. However, it is not yet clear whether there is direct progression of low-grade superficial tumours to become invasive (a Jeckell and Hyde scenario) or whether in those patients who apparently progress from one form of the disease to the other, different tumour clones are involved and that the two tumour groups are mutually exclusive ('chalk and cheese'). If the latter is true, then attempts to identify molecular markers to predict progression of low-grade superficial bladder tumours may be fruitless. Similarly, it is not clear whether other subgroups of tumours exist that arise via different molecular pathways. There is now a large amount of molecular information about bladder cancer that facilitates examination of these possibilities. Some recent studies provide evidence for the existence of at least one further group of tumours, high-grade superficial papillary tumours, which may develop via a distinct molecular pathway. Patients with such tumours do show increased risk of disease progression and for these there may exist a real progression continuum from non-invasive to invasive. If this is the case, definition of the molecular signature of this pathway and improved understanding of the biological consequences of the events involved will be pivotal in disease management. PMID- 16352617 TI - Significance of COX-2 expression in human esophageal squamous cell carcinoma. AB - Cyclooxygenase-2 (COX-2) is well established to play an important role in the tumorigenesis of a variety of human cancers; however, the function of COX-2 in the development of esophageal squamous cell carcinoma (ESCC) remains less clear. Here, we determined, first, the pattern of COX-2 expression in normal esophageal mucosa, dysplasia, carcinoma in situ (CIS) and invasive SCC. Immunohistochemical analysis showed that, while COX-2 was weakly expressed, if at all, in normal squamous epithelium, strong COX-2 expression was detected as early as the stage of dysplasia and frequently in 20 of 26 (77%) CIS and 86 of 111 (77%) invasive SCC. Upregulation of COX-2 in ESCC was found to be significantly associated with tumor progression (R = 0.493, P < 0.01). Further, treatment of human ESCC cell lines (KYSE450 and KYSE510) with NS-398, a COX-2 specific chemical inhibitor, suppressed the production of prostaglandin E2 (PGE2) and induced cell growth inhibition, cell cycle arrest at the G1-S checkpoint, and the expression of cyclin-dependent kinase inhibitors p21waf1/cip1 and p27kip1. Finally, knockdown expression of COX-2 in KYSE450 cells by a specific COX-2 siRNA dramatically inhibited PGE2 production, cell growth and, more importantly, colony formation and tumorigenesis in nude mice. Together, this study suggested that COX-2 may be involved in an early stage of squamous cell carcinogenesis of the esophagus and has a non-redundant role in the regulation of cellular proliferation and tumorigenesis of esophageal epithelial cells. PMID- 16352618 TI - A crucial role of Nrf2 in in vivo defense against oxidative damage by an environmental pollutant, pentachlorophenol. AB - Our goal was to elucidate roles of Nrf2 in in vivo defense against pentachlorophenol (PCP), an environmental pollutant and hepatocarcinogen in mice. We examined oxidative stress and cell proliferation, along with other hepatotoxicological parameters, in the livers of nrf2-deficient (wild:+/+, heterozygous:+/-, homozygous:-/-) animals fed PCP in their diet at doses of 0, 150, 300, 600, or 1200 ppm for 4 weeks. For measurement of methoxyresorufin-O demethylase (CYP 1A2), NAD(P):quinone oxidoreductase 1 (NQO1), and UDP glucuronosyltransferase (UDP-GT), an additional study was performed with all but the 150-ppm dose. Significant elevation of 8-hydroxydeoxyguanosine (8-OH-dG) levels in the liver DNA was observed only in -/- mice treated with PCP at 1200 ppm. Levels of thiobarbituric-acid-reactive substances (TBARS) were also raised significantly compared to those of the relevant +/+ mice. Bromodeoxyuridine labeling indices (BrdU-LIs) of hepatocytes in -/- mice were significantly higher at all doses than those in the relevant +/+ mice. Relative liver weights were unchanged in mice lacking Nrf2, whereas liver weight in +/+ and +/- mice was increased. Significant elevations of serum ALP activity, but not ALT and AST activity, occurred at 600 ppm and above in -/- mice compared to the relevant +/+ mice. Histopathologically, centrilobular hepatocyte necrosis was severe in the -/ mice that received 600 ppm. Although CYP 1A2 activity was elevated in all treated mice, increases in NQO1 levels and UDP-GT activities did not occur only in -/- mice. These data suggest that Nrf2 plays a key role in prevention of PCP induced oxidative stress and cell proliferation. PMID- 16352619 TI - Kinetics of genistein and its conjugated metabolites in pregnant Sprague-Dawley rats following single and repeated genistein administration. AB - Diets high in soy-based products are well known for their estrogenic activity. Genistein, the predominant phytoestrogen present in soy, is known to interact with estrogen receptors (ER) alpha and beta and elicits reproductive effects in developing rodents. In the rat, genistein is metabolized predominantly to glucuronide and sulfate conjugates, neither of which is capable of activating ER. Therefore, it is critical to understand the delivery of free and conjugated genistein across the placenta to the fetus following maternal genistein exposure such that the potential fetal exposure to free genistein can be assessed. Genistein (4 or 40 mg/kg) was administered to pregnant Sprague-Dawley rats by oral gavage daily from gestation day (GD) 5 through 19 or on GD 19 alone. Maternal and GD 19 fetal tissues were collected 0.5, 1, 2, 4, 6, 8, 12, and 24 h following administration of the final dose on GD 19. Concentrations of genistein, genistein glucuronide, and genistein sulfate were quantitated by LC-MS/MS. In maternal plasma, genistein glucuronide was the predominant metabolite. In the fetal plasma, genistein glucuronide and genistein sulfate were the primary metabolites. Genistein levels in maternal and fetal plasma were much lower than its conjugates. The concentration of genistein in placental tissue was higher than either conjugate. Fetal concentrations of unconjugated genistein following administration of 40 mg/kg were above the EC50 for ERbeta activation. Repeated administration of 40 mg/kg genistein resulted in minor changes in genistein kinetics in the pregnant rat compared to single administration of the same dose. These data suggest that conjugated forms of genistein are not transported across the placenta. High placental concentrations of genistein indicate the placenta is a potential target organ for genistein action during gestation. PMID- 16352620 TI - Synergistic interactions between commonly used food additives in a developmental neurotoxicity test. AB - Exposure to non-nutritional food additives during the critical development window has been implicated in the induction and severity of behavioral disorders such as attention deficit hyperactivity disorder (ADHD). Although the use of single food additives at their regulated concentrations is believed to be relatively safe in terms of neuronal development, their combined effects remain unclear. We therefore examined the neurotoxic effects of four common food additives in combinations of two (Brilliant Blue and L-glutamic acid, Quinoline Yellow and aspartame) to assess potential interactions. Mouse NB2a neuroblastoma cells were induced to differentiate and grow neurites in the presence of additives. After 24 h, cells were fixed and stained and neurite length measured by light microscopy with computerized image analysis. Neurotoxicity was measured as an inhibition of neurite outgrowth. Two independent models were used to analyze combination effects: effect additivity and dose additivity. Significant synergy was observed between combinations of Brilliant Blue with L-glutamic acid, and Quinoline Yellow with aspartame, in both models. Involvement of N-methyl-D-aspartate (NMDA) receptors in food additive-induced neurite inhibition was assessed with a NMDA antagonist, CNS-1102. L-glutamic acid- and aspartame-induced neurotoxicity was reduced in the presence of CNS-1102; however, the antagonist did not prevent food color-induced neurotoxicity. Theoretical exposure to additives was calculated based on analysis of content in foodstuff, and estimated percentage absorption from the gut. Inhibition of neurite outgrowth was found at concentrations of additives theoretically achievable in plasma by ingestion of a typical snack and drink. In addition, Trypan Blue dye exclusion was used to evaluate the cellular toxicity of food additives on cell viability of NB2a cells; both combinations had a straightforward additive effect on cytotoxicity. These data have implications for the cellular effects of common chemical entities ingested individually and in combination. PMID- 16352621 TI - Does leptin contribute to uraemic cachexia? PMID- 16352622 TI - Investigation of coronary artery calcification and stenosis by coronary angiography (CAG) in haemodialysis patients. PMID- 16352623 TI - Effect of iodinated contrast agents on residual renal function in PD patients. AB - BACKGROUND: Residual renal function (RRF) is an important predictor of outcome in peritoneal dialysis (PD) patients. Although increasing emphasis has been placed on preserving RRF, the nephrotoxicity associated with contrast medium administration in PD patients remains a controversial issue. In the present prospective study, we evaluated the evolution of RRF 2 weeks after iodinated contrast medium administration (ICMA) in a group of stable PD patients, and compared it with that in a non-treated control group of stable PD subjects. METHODS: The study was conducted from January 2003 to October 2004. RRF was quantified by the average of 24 h urinary urea and creatinine clearance and peritoneal creatinine clearance (PcrCl) were analyzed, the levels of which were analysed prior to and 2 weeks following ICMA in 36 PD patients and also assessed at the same time points in a group of 36 PD non-ICMA control subjects, matched according to RRF characteristics. Two weeks following ICMA, the values for RRF, daily urine volume and PcrCl were assessed against those at baseline, and the evolution of RRF was compared between the two groups. In the ICMA group, this study was performed with adequate pre-hydration and a minimum dose of contrast medium. RESULTS: Compared with baseline values, RRF, daily urine volume and PcrCl were not found to be significantly different 2 weeks after ICMA (7.0+/-4.3 vs 7.2+/-4.3 ml/min/1.73 m(2), P = 0.12; 1324+/-696 vs 1360+/-755 ml/day, P = 0.5; and 41.1+/-9 vs 40.6+/-9 l/week/1.73 m(2), P = 0.6, respectively). Following ICMA, variations in RRF and daily urine volume were found to be comparable with those of the control group (0.1+/-0.5 vs 0.1+/-0.5 ml/min/1.73 m(2), P = 0.9; 36+/-440 vs 40+/-493 ml/day, P = 0.8, respectively). CONCLUSION: In this study, 2 weeks following ICMA, no accelerated decline in RRF was determined in stable PD patients with adequate pre-hydration, i.e. subjects treated under optimal circumstances compared with the control group. PMID- 16352624 TI - First night effect for polysomnographic data in children and adolescents with suspected sleep disordered breathing. AB - AIMS: To assess the presence of a first night effect (FNE) in children and adolescents and to examine if a single night polysomnography (PSG) is sufficient for diagnosing obstructive sleep apnoea syndrome (OSAS). METHODS: Prospective case study of 70 patients (group 1: 2-6 years, n = 22; group 2: 7-12 years, n = 32; group 3: 13-17 years, n = 16) referred for OSAS. Diagnostic criteria for OSAS: one or more of the following: (1) obstructive apnoea index (OAI) > or =1; (2) obstructive apnoea hypopnoea index (oAHI) > or =2; (3) SaO2 < or =89% in association with obstruction. RESULTS: In all age groups, but mainly in the oldest children, REMS increased during the second night, mainly at the expense of stage 2 sleep. The first night PSG correctly identified OSAS in 86%, 91%, and 100% of the children for groups 1, 2, and 3 respectively. This represents 9% false negatives for OSAS when only the first night PSG was used. All cases missed had mild OSAS, except for one with oAHI >5 on night 2. There were also seven patients with OSAS on night 1 but with a normal PSG on night 2: all had oAHI <5. CONCLUSION: There is a FNE in children and adolescents. A single night PSG is sufficient for diagnosing OSAS, but in cases with a suggestive history and examination and with a negative first night, a second night study might be advisable. PMID- 16352625 TI - Isotonic is better than hypotonic saline for intravenous rehydration of children with gastroenteritis: a prospective randomised study. AB - AIMS: To determine whether the risk of hyponatraemia in children with gastroenteritis receiving intravenous (IV) fluids is decreased by the use of 0.9% saline. METHODS: A prospective randomised study was carried out in a tertiary paediatric hospital. A total of 102 children with gastroenteritis were randomised to receive either 0.9% saline + 2.5% dextrose (NS) or 0.45% saline + 2.5% dextrose (N/2) at a rate determined by their treating physician according to hospital guidelines and clinical judgement. Plasma electrolytes, osmolality, and plasma glucose were measured before (T(0)) and 4 hours after (T(4)) starting IV fluids, and subsequently if clinically indicated. Electrolytes and osmolality were measured in urine samples. Results were analysed according to whether children were hyponatraemic (plasma sodium <135 mmol/l) or normonatraemic at T(0). RESULTS: At T(0), mean (SD) plasma sodium was 135 (3.3) mmol/l (range 124 142), with 37/102 (36%) hyponatraemic. At T(4), mean plasma sodium in children receiving N/2 remained unchanged in those initially hyponatraemic (n = 16), but fell 2.3 (2.2) mmol/l in the normonatraemic group. In contrast, among children receiving NS, mean plasma sodium was 2.4 (2.0) mmol/l higher in those hyponatraemic at baseline (n = 21) and unchanged in the initially normonatraemic children. In 16 children who were still receiving IV fluids at 24 hours, 3/8 receiving N/2 were hyponatraemic compared with 0/8 receiving NS. No child became hypernatraemic. CONCLUSIONS: In gastroenteritis treated with intravenous fluids, normal saline is preferable to hypotonic saline because it protects against hyponatraemia without causing hypernatraemia. PMID- 16352626 TI - Delayed detection of cleft palate: an audit of newborn examination. AB - AIMS: To identify prevalence of delayed detection of cleft palate, and associated factors that could lead to improved identification at neonatal clinical examination. METHODS: Audit of hospital notes, parental questionnaire incorporating open ended questions, and telephone questionnaire of junior doctors in the referring hospitals incorporating fixed choice questions. RESULTS: Of 344 cleft palate patients without cleft lip or submucous cleft palate, the day the cleft was detected was recorded in 92%. Delayed detection, after the first day, was 28% overall, distributed as 37% with isolated cleft palate and 23% with syndromic cleft palate. Narrow V shaped clefts were more likely to be delayed in detection compared with broad U shaped clefts, as were soft palate clefts compared with hard palate clefts. Five with isolated cleft palates were not detected until after the first year. Babies born at home were unlikely to be detected on day 1. Symptoms were significantly increased in the delayed detection group for feeding problems and nasal regurgitation. A telephone questionnaire of trainee paediatricians in referring units revealed that digital examination was more commonly practised than visual inspection, and few recalled receiving specific instruction on examination of the palate. CONCLUSION: Delayed detection of cleft palate was not uncommon, and the features of those more likely to be missed suggested digital examination was related. Trainee doctors and midwives should be instructed to inspect visually using a light and tongue depressor, then digitally if submucous cleft palate is suspected. PMID- 16352627 TI - Changing rooms: time for a makeover. AB - This study explores the concerns expressed by primary school children and their parents about changing arrangements for PE at school. In 39% of schools pupils had expressed anxieties. Are traditional arrangements where children get changed for PE in mixed sex classrooms still appropriate in view of the younger age at which children are now starting puberty? PMID- 16352628 TI - Peptide-induced immune protection of CD8+ T cell-deficient mice against Friend retrovirus-induced disease. AB - CD8+ CTLs and virus-neutralizing antibodies have been associated with spontaneous and vaccine-induced immune control of retroviral infections. We previously showed that a single immunization with an env gene-encoded CD4+ T cell epitope protected mice against fatal Friend retrovirus infection. Here, we analyzed immune cell components required for the peptide-induced anti-retroviral protection. Mice lacking CD8+ T cells were nevertheless protected against Friend virus infection, while mice lacking B cells were not. Virus-producing cells both in the spleen and bone marrow decreased rapidly in their number and became undetectable by 4 weeks after infection in the majority of the peptide-immunized animals even in the absence of CD8+ T cells. In the vaccinated animals the production and class switching of virus-neutralizing and anti-leukemia cell antibodies were facilitated; however, virus-induced erythroid cell expansion was suppressed before neutralizing antibodies became detectable in the serum. Further, the numbers of virus-producing cells in the spleen and bone marrow in the early stage of the infection were smaller in the peptide-immunized than in unimmunized control mice in the absence of B cells. Thus, peptide immunization facilitates both early cellular and late humoral immune responses that lead to the effective control of the retrovirus-induced disease, but CD8+ T cells are not crucial for the elimination of virus-infected cells in the peptide-primed animals. PMID- 16352629 TI - HIV-1 burden influences host response to co-infection with Neisseria gonorrhoeae in vitro. AB - There is considerable evidence that co-infection with the sexually transmitted pathogen Neisseria gonorrhoeae (Gc) can increase the likelihood of both transmitting and acquiring HIV-1 worldwide. However, less information is available on how host immune response to co-infection differs with immune response to HIV-1 infection alone. To evaluate HIV-1 burden effects on host response to co-infection with Gc, we performed gene-expression profiling of human PBMCs infected over a broad range of viral titers (HIV-1 series) and upon exposure to a single infectious dose of Gc (HIV-1/Gc series). The transcriptional profiles differed substantially between each series (P < 0.0001). Major shifts in the transcriptional landscape were identified in contour plots based on fold stimulation and hierarchical clustering. Prominent regions of transcriptional activity were evaluated for statistical enrichment to identify up-regulated pathways associated with immune response, infection and T-cell stimulation. Notably, gene enrichment was dependent on HIV-1 burden and shifted during co infection to reveal a disproportionate effect on lymphocyte signaling, apoptosis and proteasome activity. Further evaluation of these findings may help to better understand the role of viral burden in defining cellular contribution to host immune response upon co-infection with secondary sexually transmitted pathogens. PMID- 16352630 TI - TRAF1 regulates Th2 differentiation, allergic inflammation and nuclear localization of the Th2 transcription factor, NIP45. AB - We have previously reported that tumor necrosis factor receptor-associated factor 1 (TRAF1), an intracellular protein, which binds to a range of molecules, including tumor necrosis factor (TNF) receptor family members, regulates TNF induced NF-kappaB and AP-1 signaling as well as TCR-triggered proliferative responses in T cells. In order to define the role of TRAF1 in Th cell differentiation, we analyzed the responses of TRAF1-/- T cells following TCR activation. Stimulation of TRAF1-/- T cells by antigen resulted in significantly increased expression of the Th2 cytokines (IL-4, IL-5 and IL-13) compared with wild-type (WT) controls. The Th2 bias of TRAF1-/- T cells is T lymphocyte intrinsic, since naive CD4+CD62L+ TRAF1-/- T cells activated with CD3/CD28 produced elevated levels of Th2 cytokines. Consistent with these observations in cultured T cells, TRAF1-/- T cells induced enhanced Th2 responses in vivo. Transfer of ovalbumin (OVA)-immune TRAF1-/- T cells into naive WT recipients conferred significantly more intense pulmonary inflammation and higher airway hyperresponsiveness following inhaled OVA challenge than did transfer of OVA immune WT T cells. Biochemical analysis of TRAF1-/- T cells revealed that they have elevated nuclear expression of NFAT-interacting protein (NIP45), a Th2 cell associated transcription factor known to potentiate NFATp-driven IL-4 expression. In further experiments, we demonstrated that TRAF1 associates with a fraction of NIP45 in the cytoplasm and prevents its translocation to the nucleus. Taken together these results suggest that TRAF1 may limit the induction of Th2 responses by decreasing NIP45 concentration to the nucleus and thereby down regulating the expression of NIP45-dependent IL-4 gene transcription. PMID- 16352631 TI - Granulocyte colony-stimulating factor promotes tumor angiogenesis via increasing circulating endothelial progenitor cells and Gr1+CD11b+ cells in cancer animal models. AB - Recombinant granulocyte colony-stimulating factor (G-CSF) is used for cancer patients with myelosuppression induced by chemotherapy. G-CSF has been reported to progress tumor growth and angiogenesis, but the precise mechanism of tumor angiogenesis activated by G-CSF has not been fully clarified. N-terminal-mutated recombinant human G-CSF administration increased WBCs and neutrophils in peripheral blood and reduced bone marrow stromal cell-derived factor-1 in mice, indicating its biological relevance. Mice were inoculated with Lewis lung carcinoma cells (LLCs) or KLN205 cells and treated with G-CSF. G-CSF accelerated tumor growth and intratumoral vessel density, while it did not accelerate proliferation of LLCs, KLN205 cells or human umbilical vein endothelial cells in vitro. In the absence of tumors, G-CSF did not increase circulating cells that displayed phenotypic characteristics of endothelial progenitor cells (EPCs). In the presence of tumors, G-CSF increased circulating EPCs. In addition, G-CSF treatment increased immune suppressor and endothelial cell-differentiating Gr1+CD11b+ cells in tumor-bearing mice. We conclude that G-CSF promotes tumor growth by activating tumor angiogenesis via increasing circulating EPCs and Gr1+CD11b+ cells in cancer animal models. PMID- 16352632 TI - Consumer-directed health insurance: the next generation. Interview by James C Robinson. AB - Aetna has led the transition of the commercial health insurance industry from a focus on managed care to a focus on "consumer-driven" products that offer more information while demanding higher cost sharing from enrollees. But most health care costs are incurred by very sick patients who have spent beyond their cost sharing limits and are not directly affected by these financial incentives. Aetna CEO Jack Rowe discusses his firm's initiatives to moderate the cost of care for ill as well as healthy patients, including providing information on provider prices, contracting for narrow physician specialty networks, and promoting disease management and case management programs. PMID- 16352633 TI - Serum cytokine profiles and their correlations with disease activity in Takayasu's arteritis. AB - OBJECTIVE: To investigate serum profiles of inflammatory cytokines in patients with Takayasu's arteritis (TA) and to determine their correlations with disease activity of TA. METHODS: Forty-nine patients with TA and 12 age- and sex-matched controls were studied. Blood samples were obtained and were divided into active and stable disease groups. Paired blood samples were available in 19 patients at the active stage before treatment and at the remitted stage after treatment. Serum tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-6, IL-12 and IL-18 levels were determined by enzyme-linked immunosorbent assay. RESULTS: Serum TNF-alpha, IL-6 and IL-18 levels of patients with TA were significantly higher than those of controls (P<0.05), but IFN-gamma and IL-12 levels were not. Serum IL-6 and IL-18 levels were significantly higher in the active disease group than in the stable disease group (P<0.05), but the levels of TNF-alpha were not different between the groups. In the 19 patients with paired samples, serum IL-18 levels at the remitted stage after treatment were significantly decreased compared with the active stage before treatment (P<0.001). The changes in IL-18 levels between active and remitted stages correlated well with changes in erythrocyte sedimentation rate (P<0.001). CONCLUSION: Serum IL-18 and IL-6 levels were elevated in patients with TA, especially in those with active disease. Serum IL-18 levels correlated well with disease activity of TA. These results suggest that IL-6 and IL-18 might contribute to the pathogenesis of TA and that IL-18 could be a useful marker for monitoring disease activity of TA. PMID- 16352635 TI - Skin sensitivity to UVB irradiation in systemic lupus erythematosus is not related to the level of apoptosis induction in keratinocytes. AB - OBJECTIVES: Accumulation of apoptotic cells has been suggested to be involved in the pathogenesis of systemic lupus erythematosus (SLE). As sunlight exposure is one of the factors that can trigger disease activity, we hypothesized that UV light may induce increased numbers of apoptotic cells in SLE. METHODS: Fourteen SLE patients and 16 controls were irradiated with UVB to determine their minimal erythemal dose (MED). Subsequently, skin was irradiated with 1 MED and 2 MED, respectively, and after 24 h skin biopsies were analysed immunohistologically for the number of apoptotic cells and presence of pyknotic nuclear debris. RESULTS: MED was significantly decreased in SLE patients and the presence of decreased MED was associated with a history of butterfly rash. Decreased MED was not related to other skin-related ACR criteria or to autoantibody specificities. No differences were detected in the numbers of apoptotic keratinocytes between patients and controls or in the amount of pyknotic nuclear debris following 1 and 2 MED irradiation, respectively. Absolute UVB doses were correlated with the number of apoptotic keratinocytes; dose-responses did not differ significantly between patients and controls. CONCLUSIONS: Increased sensitivity of SLE patients to UVB, although associated with a history of malar rash, is not related to increased induction of apoptosis or increased levels of secondary necrosis in the skin. Thus, compared with controls, UVB-induced apoptosis is not increased in SLE patients under physiological conditions. PMID- 16352636 TI - Shortening infusion times for infliximab administration. PMID- 16352637 TI - Post-interventional immunosuppressive treatment and vascular restenosis in Takayasu's arteritis. AB - OBJECTIVE: To investigate the outcome of vascular interventions and the effect of post-interventional immunosuppressive treatment on the occurrence of vascular restenosis in patients with Takayasu's arteritis (TA). METHODS: Forty-two patients with TA who had undergone vascular intervention and had serial angiographies before and after intervention were enrolled. The demographic and clinical data were collected at the time when the interventions were performed, and the intervention modalities and post-interventional medical treatments were evaluated. RESULTS: Sixty-three interventions were performed in 42 patients. Twenty (31.7%) interventions restenosed 24.0 +/- 21.9 months after intervention; the likelihood decreasing as time passed. Estimates of arterial patency after intervention were 90.1% at 1 yr, 75.5% at 2 yr, 68.4% at 3 yr, 61.6% at 5 yr and 49.3% at 10 yr. According to the log rank test, interventions that were performed during the stable stage of the disease (P = 0.039) and those that were followed by treatment with glucocorticoids and immunosuppressive agents (P = 0.044) were independent variables for the maintenance of arterial patency. Their hazard ratios were 0.30 and 0.41, respectively. CONCLUSION: Restenosis occurred in 31.7% of TA patients after intervention. A lower restenosis rate was observed when the vascular interventions were performed at the stable stage and when post interventional immunosuppressive treatment was implemented. PMID- 16352638 TI - The impact of Picture Archiving and Communication Systems (PACS) implementation in rheumatology. PMID- 16352639 TI - Copying correspondence to patients. PMID- 16352640 TI - Detection and initial characterization of synovial lining fragments in synovial fluid. AB - OBJECTIVE: Free fragments of synovium have occasionally been seen in synovial fluid but have not been studied systematically. We wished to establish a method for the reliable detection of these fragments in joint and bursa effusions and begin to characterize them by histochemical and immunohistochemical methods. METHODS: Cell smears, wet drop preparations and cytospins were prepared from 39 consecutive joint and bursa effusions. Paraffin cell blocks were prepared from a subset. Analysis encompassed standard and polarized light microscopy, histochemistry, immunohistochemistry and transmission electron microscopy (EM). Synovial biopsy tissue from one different patient was examined for comparison. RESULTS: Tissue fragments were not seen in Wright-stained cell smears and only rarely in wet drop preparations. In contrast, variously sized fragments with the histological appearance of hyperplastic synovial lining were detected in ethanol fixed, haematoxylin/eosin-stained cytospins from bursitis and all arthropathies studied [17/24 (71%) of non-inflammatory and 12/15 (80%) of inflammatory specimens]. Immunostaining revealed CD68 expression in a subset of cells in a pattern characteristic of hyperplastic synovial lining. Juxtaposed cells with morphological features of macrophage-like and fibroblast-like synoviocytes were seen by EM. CONCLUSIONS: Synovial lining fragments can be detected in effusions from diverse arthropathies and bursitis. They maintain important properties of the synovial lining and can be analysed by immunohistochemistry. They may afford the opportunity to study a relatively pure preparation of synovial lining cells without the need for cell culture, and to evaluate their possible role in augmenting or perpetuating synovitis or joint damage. PMID- 16352642 TI - Sequential synovial fluid sampling suggests plasma and synovial fluid IL-6 vary independently in rheumatoid arthritis. PMID- 16352643 TI - Renal tubular acidosis, arthritis and autoantibodies: primary Sjogren's syndrome in childhood. PMID- 16352644 TI - Successful treatment of severe Kikuchi's disease with intravenous immunoglobulin. PMID- 16352645 TI - Targeted calretinin expression in granule cells of calretinin-null mice restores normal cerebellar functions. AB - Ca2 binding proteins such as calretinin, characterized by the presence of EF-hand motifs that bind Ca2+ ions, are involved in the shaping of intraneuronal Ca2+ fluxes. In the cerebellar cortex, information processing tightly relies on variations in intracellular Ca2+ concentration in Purkinje and granule cells. Calretinin-deficient (Cr-/-) mice present motor discoordination, suggesting cellular and network cerebellar dysfunctions. To determine the cell specificity of these alterations, we constructed transgenic Cr-/- mice exhibiting a selective reexpression of calretinin in granule cells through the promoter function of the GABAA receptor alpha6 subunit gene. Normal granule cell excitability and wild type Purkinje cell firing behavior in awake mice were restored while the emergence of high-frequency oscillations was abolished. Behavioral analysis of these calretinin-rescue mice revealed that normal motor coordination was restored as compared with Cr-/- mice. These results demonstrate that calretinin is required specifically in granule cells for correct computation in the cerebellar cortex and indicate that the finetuning of granule cell excitability through regulation of Ca2+ homeostasis plays a crucial role for information coding and storage in the cerebellum. PMID- 16352646 TI - STAT4- and STAT6-signaling molecules in a murine model of multiple sclerosis. AB - Epidemiological studies suggest that an environmental factor (possibly a virus) acquired early in life may trigger multiple sclerosis (MS). The virus may remain dormant in the central nervous system but then becomes activated in adulthood. All existing models of MS are characterized by inflammation or demyelination that follows days after virus infection or antigen inoculation. While investigating the role of CD4+ T cell responses following Theiler's virus infection in mice deficient in STAT4 or STAT6, we discovered a model in which virus infection was followed by demyelination after a very prolonged incubation period. STAT4-/- mice were resistant to demyelination for 180 days after infection, but developed severe demyelination after this time point. Inflammatory cells and up-regulation of Class I and Class II MHC antigens characterized these lesions. Virus antigen was partially controlled during the early chronic phase of the infection even though viral RNA levels remained high throughout infection. Demyelination correlated with the appearance of virus antigen expression. Bone marrow reconstitution experiments indicated that the mechanism of the late onset demyelination was the result of the STAT4-/- immune system. Thus, virus infection of STAT4-/- mice results in a model that may allow for dissection of the immune events predisposing to late-onset demyelination in MS. PMID- 16352647 TI - Disparate evolution of prion protein domains and the distinct origin of Doppel- and prion-related loci revealed by fish-to-mammal comparisons. AB - Prions result from the misfolding and selective accumulation of the host-encoded prion protein (PrP) in the brain. Despite intensive research on mammalian models, basic questions about the biological role of PrP and the evolutionary origin of prion disease remain unanswered. Following our previous identification of novel fish PrP homologues, here we generated new fish PrP sequences and performed genomic analysis to demonstrate the existence of two homologous PrP loci in bony fish, which display extensive molecular variation and are highly expressed in adult and developing fish brains. The fish PrP genomic regions contain PrP related loci directly downstream of each PrP locus, suggesting an independent origin of prion-related proteins in fish and mammals. Our structural prediction analysis uncovers a conserved molecular "bauplan" for all vertebrate PrPs. The C- and N-terminal protein domains have evolved independently from one another, the former having retained its basic globular structure despite high sequence divergence and the latter having undergone differential expansion-degeneration cycles in its repetitive domains. Our evolutionary analysis redefines fundamental concepts on the functional significance of PrP domains and opens up new possibilities for the experimental analysis of prion misfolding and neurodegeneration in a non-mammalian model like the zebrafish. PMID- 16352648 TI - Subchronic alpha- and beta-adrenergic regulation of cardiac gap junction protein expression. AB - Gap junction channels are essential for intercellular electrical communication in the heart. The most important cardiac gap junction proteins are connexin43 (predominantly) (Cx43), connexin40 (Cx40), and in early developmental stages connexin45. Since catecholamines play an important role in cardiac (patho)physiology, we wanted to elucidate whether catecholamines may affect expression of Cx43 and Cx40. Cultured neonatal rat cardiomyocytes were exposed for 24 h to increasing concentrations of noradrenaline (1-10000 nM) (physiological agonist at alpha and beta-adrenoceptors), resulting in significantly increased Cx43-expression, while Cx40 was unaffected. In further experiments cells were incubated with either phenylephrine (alpha-adrenergic agonist) or isoproterenol (beta-adrenergic agonist) (0.1-1000 nM) for 24 h. Both catecholamines lead to a concentration-dependent increase in Cx43 protein and mRNA expression (EC50: 10-20 nM). Inhibition experiments showed that the phenylephrine effect was transduced via PKC, while the isoproterenol effect was mediated by PKA. Dual whole-cell voltage clamp demonstrated that increased Cx43 expression was accompanied by significant increases in gap junction current. In additional in vivo experiments, adult rats were subjected to 24-h infusion of isoproterenol or phenylephrine showing again significant increase in Cx43 but not Cx40. Adrenergic stimulation of cardiomyocytes can enhance Cx43 expression thereby increasing cellular coupling, indicating a possible role for catecholamines in the regulation of cardiac gap junction expression in cardiac disease. PMID- 16352649 TI - The antibiotic and DNA-transfecting peptide LAH4 selectively associates with, and disorders, anionic lipids in mixed membranes. AB - The histidine-rich amphipathic peptide LAH4 has antibiotic and DNA delivery capabilities. The peptide has a strong affinity for anionic lipids found in the outer membrane of bacterial membranes. A role for anionic lipids in release of cationic plasmid-containing complexes has been proposed previously, and disruption of membrane asymmetry and presentation of phosphatidylserine (PS) in the membrane outer leaflet is a general feature observed in diseased mammalian cells. Therefore, to understand the peptide-lipid interactions in more detail, solid-state NMR experiments on model membranes have been performed. 31P MAS NMR on mixed phosphatidylcholine (PC)/PS and PC/phosphatidylglycerol (PG) membranes has been used to demonstrate a strong interaction between LAH4 and anionic lipids. By using deuterated lipids and wide-line 2H NMR when probing lipid chain order, it is demonstrated that LAH4 preferentially interacts with PS over PC and effectively disorders the anionic PS lipid fatty acyl chains. In addition, we demonstrate that the efficiency of gene transfer in vitro to different cell lines is closely related to the degree of disruption of PS acyl chains for four isomers of LAH4. This work suggests a mechanism of selective destabilization by LAH4 of anionic lipids in the membranes of cells during transfection with implications for nucleic acid delivery in vivo. PMID- 16352650 TI - Variant isoforms of CD44 are P- and L-selectin ligands on colon carcinoma cells. AB - The initial selectin-dependent events that mediate tumor cell tethering to platelets, leukocytes, and vascular endothelium can regulate the extravasation and colonization of metastatic cells into distant tissues. Little is known, however, about the identity of selectin counter-receptors on tumor cells, which facilitate the metastatic process. To address this issue, we performed SDS-PAGE analysis of membrane proteins, metabolic inhibition studies, blot rolling assays, and cell-free flow-based adhesion experiments using microbeads coated with CD44 immunoprecipitated from carcinomas and purified selectins as substrate. Here, we demonstrate that variant isoforms of CD44 (CD44v) on LS174T colon carcinoma cells possess P-/L-/E-selectin binding activity, in contrast to the standard isoform of CD44 (CD44s) on hematopoietic-progenitor cells (HPCs), which is primarily an L-/E selectin ligand. Moreover, the selectin-binding determinants on CD44v from LS174T cells are sialofucosylated structures displayed on O-linked glycans, akin to those on P-selectin glycoprotein ligand-1, but distinct from the HECA-452 reactive N-glycans on CD44s expressed on HPCs. Using flow-based adhesion assays, we systematically characterize shear-dependent LS174T CD44 vs. HL60 CD44s adhesion to E-/P-/L-selectin. The novel finding that CD44v are selectin ligands offers a unifying perspective on the apparent enhanced metastatic potential associated with tumor cell CD44v overexpression and the critical role of selectins in metastasis. PMID- 16352651 TI - Plasmid electrotransfer of eye ciliary muscle: principles and therapeutic efficacy using hTNF-alpha soluble receptor in uveitis. AB - Due to its small size and particular isolating barriers, the eye is an ideal target for local therapy. Recombinant protein ocular delivery requires invasive and painful repeated injections. Alternatively, a transfected tissue might be used as a local producer of transgene-encoded therapeutic protein. We have developed a nondamaging electrically mediated plasmid delivery technique (electrotransfer) targeted to the ciliary muscle, which is used as a reservoir tissue for the long-lasting expression and secretion of therapeutic proteins. High and long-lasting reporter gene expression was observed, which was restricted to the ciliary muscle. Chimeric TNF-alpha soluble receptor (hTNFR-Is) electrotransfer led to elevated protein secretion in aqueous humor and to drastic inhibition of clinical and histological inflammation scores in rats with endotoxin-induced uveitis. No hTNFR-Is was detected in the serum, demonstrating the local delivery of proteins using this method. Plasmid electrotransfer to the ciliary muscle, as performed in this study, did not induce any ocular pathology or structural damage. Local and sustained therapeutic protein production through ciliary muscle electrotransfer is a promising alternative to repeated intraocular protein administration for a large number of inflammatory, degenerative, or angiogenic diseases. PMID- 16352652 TI - Quality classification of tandem mass spectrometry data. AB - Peptide identification by tandem mass spectrometry is an important tool in proteomic research. Powerful identification programs exist, such as SEQUEST, ProICAT and Mascot, which can relate experimental spectra to the theoretical ones derived from protein databases, thus removing much of the manual input needed in the identification process. However, the time-consuming validation of the peptide identifications is still the bottleneck of many proteomic studies. One way to further streamline this process is to remove those spectra that are unlikely to provide a confident or valid peptide identification, and in this way to reduce the labour from the validation phase. RESULTS: We propose a prefiltering scheme for evaluating the quality of spectra before the database search. The spectra are classified into two classes: spectra which contain valuable information for peptide identification and spectra that are not derived from peptides or contain insufficient information for interpretation. The different spectral features developed for the classification are tested on a real-life material originating from human lymphoblast samples and on a standard mixture of 9 proteins, both labelled with the ICAT-reagent. The results show that the prefiltering scheme efficiently separates the two spectra classes. PMID- 16352653 TI - Improved pairwise alignments of proteins in the Twilight Zone using local structure predictions. AB - MOTIVATION: In recent years, advances have been made in the ability of computational methods to discriminate between homologous and non-homologous proteins in the 'twilight zone' of sequence similarity, where the percent sequence identity is a poor indicator of homology. To make these predictions more valuable to the protein modeler, they must be accompanied by accurate alignments. Pairwise sequence alignments are inferences of orthologous relationships between sequence positions. Evolutionary distance is traditionally modeled using global amino acid substitution matrices. But real differences in the likelihood of substitutions may exist for different structural contexts within proteins, since structural context contributes to the selective pressure. RESULTS: HMMSUM (HMMSTR based substitution matrices) is a new model for structural context-based amino acid substitution probabilities consisting of a set of 281 matrices, each for a different sequence-structure context. HMMSUM does not require the structure of the protein to be known. Instead, predictions of local structure are made using HMMSTR, a hidden Markov model for local structure. Alignments using the HMMSUM matrices compare favorably to alignments carried out using the BLOSUM matrices or structure-based substitution matrices SDM and HSDM when validated against remote homolog alignments from BAliBASE. HMMSUM has been implemented using local Dynamic Programming and with the Bayesian Adaptive alignment method. PMID- 16352654 TI - Differential gene expression detection and sample classification using penalized linear regression models. AB - Differential gene expression detection and sample classification using microarray data have received much research interest recently. Owing to the large number of genes p and small number of samples n (p >> n), microarray data analysis poses big challenges for statistical analysis. An obvious problem owing to the 'large p small n' is over-fitting. Just by chance, we are likely to find some non differentially expressed genes that can classify the samples very well. The idea of shrinkage is to regularize the model parameters to reduce the effects of noise and produce reliable inferences. Shrinkage has been successfully applied in the microarray data analysis. The SAM statistics proposed by Tusher et al. and the 'nearest shrunken centroid' proposed by Tibshirani et al. are ad hoc shrinkage methods. Both methods are simple, intuitive and prove to be useful in empirical studies. Recently Wu proposed the penalized t/F-statistics with shrinkage by formally using the (1) penalized linear regression models for two-class microarray data, showing good performance. In this paper we systematically discussed the use of penalized regression models for analyzing microarray data. We generalize the two-class penalized t/F-statistics proposed by Wu to multi class microarray data. We formally derive the ad hoc shrunken centroid used by Tibshirani et al. using the (1) penalized regression models. And we show that the penalized linear regression models provide a rigorous and unified statistical framework for sample classification and differential gene expression detection. PMID- 16352655 TI - A simple method to predict protein-binding from aligned sequences--application to MHC superfamily and beta2-microglobulin. AB - MOTIVATION: The MHC superfamily (MhcSF) consists of immune system MHC class I (MHC-I) proteins, along with proteins with a MHC-I-like structure that are involved in a large variety of biological processes. beta2-Microglobulin (B2M) non-covalent binding to MHC-I proteins is required for their surface expression and function, whereas MHC-I-like proteins interact, or not, with B2M. This study was designed to predict B2M binding (or non-binding) of newly identified MhcSF proteins, in order to decipher their function, understand the molecular recognition mechanisms and identify deleterious mutations. IMGT standardization of MhcSF protein domains provides a unique numbering of the multiple alignment positions, and conditions to develop such predictive tool. METHOD: We combine a simple-Bayes classifier with IMGT unique numbering. Our method involves two steps: (1) selection of discriminant binary features, which associate an alignment position with an amino acid group; and (2) learning of the classifier by estimating the frequencies of selected features, conditionally to the B2M binding property. RESULTS: Our dataset contains aligned sequences of 806 allelic forms of 47 MhcSF proteins, corresponding to 9 receptor types and 4 mammalian species. Eighteen discriminant features are selected, belonging to B2M contact sites, or stabilizing the molecular structure required for this contact. Three leave-one-out procedures are used to assess classifier performance, which corresponds to B2M binding prediction for: (1) new proteins, (2) species not represented in the dataset and (3) new receptor types. The prediction accuracy is high, i.e. 98, 94 and 70%, respectively. Application of our classifier to lower vertebrate MHC-I proteins indicates that these proteins bind to B2M and should then be expressed on the cellular surface by a process similar to that of mammalian MHC-I proteins. These results demonstrate the usefulness and accuracy of our (simple) approach, which should apply to other function or interaction prediction problems. PMID- 16352656 TI - Comorbid phobic disorders do not influence outcome of alcohol dependence treatment. Results of a naturalistic follow-up study. AB - AIMS: Despite claims that comorbid anxiety disorders tend to lead to a poor outcome in the treatment of alcohol dependence, the few studies on this topic show conflicting results. OBJECTIVE: To test whether the outcome of treatment seeking alcohol-dependent patients with a comorbid phobic disorder is worse than that of similar patients without a comorbid phobic disorder. METHODS: The probabilities of starting to drink again and of relapsing into regular heavy drinking in (i) a group of 81 alcohol-dependent patients with comorbid social phobia or agoraphobia were compared with those in (ii) a group of 88 alcohol dependent patients without anxiety disorders in a naturalistic follow-up using Cox regression analysis. RESULTS: Adjusted for initial group differences, the hazard ratio for the association of phobic disorders with resumption of drinking was 1.05 (95% CI, 0.85-1.30, P = 0.66) and the adjusted hazard ratio for the association of phobic disorders with a relapse into regular heavy drinking was 1.02 (95% CI, 0.78-1.33, P = 0.89). CONCLUSION: The findings of this study do not confirm the idea that alcohol-dependent patients who have undergone alcohol dependence treatment are at greater risk of a relapse if they have a comorbid anxiety disorder. No differences were found in abstinence duration or time to relapse into regular heavy drinking between patients with and without comorbid phobic disorders. PMID- 16352657 TI - Effects of a web based decision aid on parental attitudes to MMR vaccination: a before and after study. AB - OBJECTIVE: To determine whether an evidence based decision aid on the measles, mumps, and rubella (MMR) vaccine changed parents' attitudes towards vaccination. DESIGN: Before and after study. SETTING: Website promoted through search engines and online parenting and health websites. PARTICIPANTS: 158 people completing online questions out of 1277 who accessed the website. INTERVENTION: Evidence based decision aid with text and graphical representation of the possible outcomes of measles, mumps, and rubella diseases compared with MMR vaccination. MAIN OUTCOME MEASURES: Attitudes towards MMR vaccination. RESULTS: Significantly more participants indicated they were "leaning towards" vaccination after using the decision aid (39% before v 55% after, P < 0.001). Compared with those who were undecided or "leaning away from" vaccination after using the decision aid, participants "leaning towards" it were more likely to strongly value its potential to protect their children from the serious side effects of disease (98% v 84%) and other children who could not be vaccinated for medical reasons (68% v 25%). Participants with an unfavourable attitude to vaccination were more likely to be very concerned about the rare side effects of vaccination (78% v 57%), have residual concerns about autism and bowel disease (78% v 27%), and anticipate guilt should their child have an adverse reaction (77% v 50%). Those with a positive attitude to vaccination after using the decision aid were also more likely to feel well informed (83% v 48%). CONCLUSIONS: A web based decision aid significantly improved parental attitudes to MMR vaccination. Residual concerns about autism and bowel disease and the rare chance of serious complications remained as attitudinal barriers to some parents. PMID- 16352658 TI - Centralspindlin regulates ECT2 and RhoA accumulation at the equatorial cortex during cytokinesis. AB - During determination of the cell division plane, an actomyosin contractile ring is induced at the equatorial cell cortex by signals from the mitotic apparatus and contracts to cause cleavage furrow progression. Although the small GTPase RhoA is known to regulate the progression, probably by controlling actin filament assembly and enhancing actomyosin interaction, any involvement of RhoA in division plane determination is unknown. In this study, using a trichloroacetic acid (TCA) fixation protocol we recently developed, we show that RhoA accumulates at the equatorial cortex before furrow initiation and continues to concentrate at the cleavage furrow during cytokinesis. We also demonstrate that both Rho activity and microtubule organization are required for RhoA localization and proper furrowing. Selective disruption of microtubule organization revealed that both astral and central spindle microtubules can recruit RhoA at the equatorial cortex. We find that centralspindlin and ECT2 are required for RhoA localization and furrowing. Centralspindlin is localized both to central spindle microtubules and at the tips of astral microtubules near the equatorial cortex and recruits ECT2. Positional information for division plane determination from microtubules is transmitted to the cell cortex to organize actin cytoskeleton through a mechanism involving these proteins. PMID- 16352659 TI - ER stress disrupts Ca2+-signaling complexes and Ca2+ regulation in secretory and muscle cells from PERK-knockout mice. AB - Disruption of protein synthesis and folding results in ER stress, which is associated with the pathophysiology of diverse diseases affecting secretory and muscle cells. Cells are protected against ER stress by activation of the unfolded protein response (UPR) that is regulated by the protein kinase PERK, which phosphorylates the translation initiation factor 2 eIF2alpha to attenuate protein synthesis. PERK-/- cells are unable to modulate ER protein load and experience high levels of ER stress. In addition to its role in protein synthesis, the ER also orchestrates many signaling events essential for cell survival, prominent among which is Ca2+ signaling. It is not known, however, whether there is a relationship between ER stress and the function of the Ca2+-signaling pathway in muscle and non-muscle cells. To directly address this question we characterized Ca2+ signaling in the secretory pancreatic and parotid acinar cells and in urinary bladder smooth muscle (UBSM) cells obtained from PERK-/- and wild-type mice. Deletion of PERK that results in high levels of ER stress, and distention and fragmentation of the ER slowed the rate of agonist-mediated Ca2+ release from the ER and reduced Ca2+-induced Ca2+ release, although IP3 production, localization of the IP3 receptors, IP3-mediated Ca2+ release, Ca(v)1.2 current and RyRs activity remained unaltered. On the other hand, ER stress disrupted the integrity of the Ca2+-signaling complexes in both secretory and UBSM cells, as revealed by markedly reduced co-immunoprecipitation of plasma membrane- and ER resident Ca2+-signaling proteins. These findings establish a relationship between the unfolding protein response, ER stress and Ca2+ signaling and highlight the importance of communication within the terminal ER-plasma membrane microdomain for propagation of the Ca2+ signal from the plasma membrane into the cell. PMID- 16352660 TI - Beta2-chimaerin provides a diacylglycerol-dependent mechanism for regulation of adhesion and chemotaxis of T cells. AB - The small GTPase Rac contributes to regulation of cytoskeletal rearrangement during chemokine-induced lymphocyte adhesion and migration in a multi-step process that is very precisely coordinated. Chimaerins are Rac1-specific GTPase activating proteins of unknown biological function, which have a canonical diacylglycerol C1-binding domain. Here we demonstrate endogenous expression of beta2-chimaerin in T lymphocytes and study the functional role of this protein in phorbol ester and chemokine (CXCL12)-regulated T-cell responses. We used green fluorescent protein-tagged beta2-chimaerin and phorbol ester stimulation to investigate changes in protein localization in living lymphocytes. Our results demonstrate that active Rac cooperates with C1-dependent phorbol ester binding to induce sustained GFP-beta2-chimaerin localization to the membrane. Subcellular distribution of GFP beta2-chimaerin in living cells showed no major changes following CXCL12 stimulation. Nonetheless Rac1-GTP levels were severely inhibited in GFP-beta2-chimaerin-expressing cells, which displayed reduced CXCL12-induced integrin-dependent adhesion and spreading. This effect was dependent on chimaerin GTPase-activating protein function and required diacylglycerol generation. Whereas beta2-chimaerin overexpression decreased static adhesion, it enhanced CXCL12-dependent migration via receptor-dependent diacylglycerol production. These studies demonstrate that beta2-chimaerin provides a novel, diacylglycerol dependent mechanism for Rac regulation in T cells and suggest a functional role for this protein in Rac-mediated cytoskeletal remodeling. PMID- 16352661 TI - Cell adhesion regulates Ser/Thr phosphorylation and proteasomal degradation of HEF1. AB - Human enhancer of filamentation 1 (HEF1), a multifunctional docking protein of the Cas family, participates in integrin and growth factor signaling pathways that regulate global cellular processes including growth, motility and apoptosis. HEF1 consists of two isoforms, p105 and p115, the larger molecular weight form resulting from Ser/Thr phosphorylation of p105HEF1. The molecular mechanisms that regulate the interconversion of the two HEF1 species as well as the function of HEF1 Ser/Thr phosphorylation are unknown. Our study reveals that cell adhesion and detachment regulate the interconversion of the two HEF1 isoforms. Experiments using various inhibitors of cytoskeletal organization indicated that disruption of actin microfilaments but not intermediate filaments or microtubules resulted in a complete conversion of p115HEF1 to p105HEF1. The conversion of p115HEF1 to p105HEF1 was prevented by inhibition of protein phosphatase 2A (PP2A), suggesting that cytoskeletal regulation of PP2A activity controlled the dephosphorylation of p115HEF1. Degradation of endogenous HEF1 was dependent on proteasomes with the p115 species of HEF1 being preferentially targeted for turnover. Dephosphorylation of HEF1 by suspending cells or disrupting actin filaments protected HEF1 from degradation. These results suggest that the adhesion dependent actin organization regulates proteasomal turnover of HEF1 through the activity of PP2A. PMID- 16352662 TI - Geldanamycin stimulates internalization of ErbB2 in a proteasome-dependent way. AB - The potent oncoprotein and receptor tyrosine kinase ErbB2 is remarkable because it resists efficient downregulation. However, ErbB2 can be downregulated by the HSP-90 inhibitor geldanamycin, but the underlying cellular mechanisms are uncertain. Apparently, delivery of ErbB2 to lysosomes, cleavage of the ErbB2 kinase domain and proteasomal activity are all processes that are involved. Using a non-invasive confocal microscopical assay allowing quantitative analysis of ErbB2 internalization in cell populations, we show that whereas ErbB2 is resistant to internalization in untreated SK-BR-3 cells, geldanamycin stimulates internalization and subsequent degradation in lysosomes. This process depends on proteasomal activity, which is a regulatory upstream event in ErbB2 internalization rather than the actual mechanism of degradation. ErbB2 can be internalized as a full-length protein, thus cleavage of the ErbB2 kinase domain is not a requirement for geldanamycin-stimulated internalization. Moreover, as shown by FRAP (fluorescence recovery after photobleaching) and electron microscopy, geldanamycin induces an increase in the amount of mobile ErbB2 and a redistribution of ErbB2 in the plasma membrane making the receptor accessible to endocytosis. Cells with most ErbB2 endocytosis also have the highest fraction of mobile ErbB2. It is concluded that geldanamycin stimulates internalization of full-length ErbB2 in a proteasome-dependent manner leading to lysosomal degradation. PMID- 16352663 TI - FGFR-1 [corrected] signaling is involved in spermiogenesis and sperm capacitation. AB - Cloning of the fibroblast growth factor receptor (FGFR) adaptor Snt-2 cDNA and the identification of FGFR-1 protein in association with sperm tails, suggested that FGFR-1 signaling was involved in either sperm tail development or function. This hypothesis was tested by the creation of transgenic mice that specifically expressed a dominant-negative variant of FGFR-1 in male haploid germ cells. Mating of transgenic mice showed a significant reduction in pups per litter compared with wild-type littermates. Further analysis demonstrated that this subfertility was driven by a combination of reduced daily sperm output and a severely compromised ability of those sperm that were produced to undergo capacitation prior to fertilization. An analysis of key signal transduction proteins indicated that FGFR-1 is functional on wild-type sperm and probably signals via the phosphatidylinositol 3-kinase pathway. FGFR-1 activation also resulted in the downstream suppression of mitogen activated protein kinase signaling. These data demonstrate the FGFR-1 is required for quantitatively and qualitatively normal spermatogenesis and has a key role in the regulation of the global tyrosine phosphorylation events associated with sperm capacitation. PMID- 16352664 TI - Nuclear protein NP60 regulates p38 MAPK activity. AB - The activation of p38alpha is mediated by its upstream kinase and associated proteins. Here we identify a new nuclear protein, NP60, which regulates the activation of p38alpha in response to sorbitol treatment. NP60 specifically binds to p38alpha, but not to JNK and ERK, in vitro and in vivo. Co-transfection of NP60 leads to the phosphorylation and activation of p38alpha, and subsequently results in the phosphorylation and activation of activating transcription factor 2. The phosphorylation of p38alpha induced by NP60 requires upstream activity of p38alpha MAP kinase, MAP kinase kinase 6 (MKK6) or MKK4. Our results indicate that NP60 mediates stress activation of p38alpha and regulates p38alpha signaling in a specific way. PMID- 16352665 TI - Effects of chronic Akt activation on glucose uptake in the heart. AB - Acute activation of the serine-threonine kinase Akt is cardioprotective and increases glucose uptake, at least in part, through enhanced expression of GLUT4 on the sarcolemma. The effects of chronic Akt activation on glucose uptake in the heart remain unclear. To address this issue, we examined the effects of chronic Akt activation on glucose uptake, glycogen storage, and relevant glucose transporters in the hearts of transgenic mice. We found that chronic cardiac activation of Akt led to a substantial increase in the rate of basal glucose uptake (P < 0.05) but blunted the response to insulin (1.9 vs. 18.1-fold increase compared with baseline) using NMR in ex vivo perfused heart. Basal glucose uptake was also increased in Akt transgenic mice in vivo (P < 0.005). These changes were associated with an increase on glycogen deposition, examined with histochemical staining, biochemical (>6-fold, P < 0.001) and in vivo radioactive (5-fold, P < 0.01) assays. Studies in chimeric hearts of female X-linked transgenic Akt mice suggested that increased glycogen deposition occurred as a cell autonomous effect of transgene expression. Interestingly, although sarcolemmal GLUT1 was not significantly altered, chronic Akt activation actually decreased plasma membrane GLUT4. Moreover, intracellular pools of GLUT1 were modestly reduced, whereas intracellular GLUT4 was substantially reduced. It seems likely that neither GLUT1 nor GLUT4 explains the increase in basal glucose uptake but that these reductions contribute to the loss of insulin responsiveness that we observed. These data demonstrate that chronic Akt activation increases basal glucose uptake and glycogen deposition while inhibiting the response to insulin. PMID- 16352666 TI - SUMO-1, human male germ cell development, and the androgen receptor in the testis of men with normal and abnormal spermatogenesis. AB - Sumoylation affects multiple cellular events, including chromatin inactivation and transcriptional repression. Our data provide the first characterization of small ubiquitin-related modifier-1 (SUMO-1) expression during human spermatogenesis by the use of high-resolution cellular SUMO-1 bioimaging. During human meiotic prophase, SUMO-1 localizes to sex chromosomes and centromeric and pericentromeric chromatin. As human spermatocytes progress toward the end of prophase in meiosis I, SUMO-1 is no longer detected within the sex body and pericentromeric heterochromatin but localizes exclusively to centromeres. SUMO-1 localization along sex chromosome axes, pseudoautosomal region, and centromeres of both chromosomes supports a role for SUMO-1 sumoylation in epigenetic events occurring over the entire sex body, e.g., meiotic sex chromosome inactivation and chromatin condensation. Centromeric SUMO-1 throughout meiotic prophase suggests a role in centromeric chromatin condensation and/or other centromere/kinetochore functions. SUMO-1 is likely involved in both facultative and constitutive heterochromatin processes in spermatocytes. Haploid round spermatids show a consistent association of SUMO-1 with centromeric clusters. During spermatid elongation, SUMO-1 localizes in the manchette perinuclear ring. Steroidogenic Leydig cells show some cytoplasmic but strong nuclear and perinuclear SUMO-1. Peritubular myoepithelial cell SUMO-1 colocalizes with centromeric heterochromatin. In epithelial Sertoli cells, when associated with centromeric heterochromatin, SUMO-1 is adjacent but not colocalized with the nucleolus. Male germ cells demonstrate no SUMO-1 nucleolar association. Human and rodent Sertoli cells consistently show an inverse correlation between androgen receptor (AR) and SUMO-1 expression and compartmentalization. Sertoli cells from certain infertile patients, however, showed greatly decreased SUMO-1 and AR. Our data suggest that human testicular SUMO-1 has specific functions in heterochromatin organization, meiotic centromere function, and gene expression. PMID- 16352667 TI - Diet and exercise reduce low-grade inflammation and macrophage infiltration in adipose tissue but not in skeletal muscle in severely obese subjects. AB - Obesity is associated with low-grade inflammation, insulin resistance, type 2 diabetes, and cardiovascular disease. This study investigated the effect of a 15 wk lifestyle intervention (hypocaloric diet and daily exercise) on inflammatory markers in plasma, adipose tissue (AT), and skeletal muscle (SM) in 27 severely obese subjects (mean body mass index: 45.8 kg/m2). Plasma samples, subcutaneous abdominal AT biopsies, and vastus lateralis SM biopsies were obtained before and after the intervention and analyzed by ELISA and RT-PCR. The intervention reduced body weight (P < 0.001) and increased insulin sensitivity (homeostasis model assessment; P < 0.05). Plasma adiponectin (P < 0.001) increased, and C-reactive protein (P < 0.05), IL-6 (P < 0.01), IL-8 (P < 0.05), and monocyte chemoattractant protein-1 (P < 0.01) decreased. AT inflammation was reduced, determined from an increased mRNA expression of adiponectin (P < 0.001) and a decreased expression of macrophage-specific markers (CD14, CD68), IL-6, IL-8, and tumor necrosis factor-alpha (P < 0.01). After adjusting for macrophage infiltration in AT, only IL-6 mRNA was decreased (P < 0.05). Only very low levels of inflammatory markers were found in SM. The intervention had no effect on adiponectin receptor 1 and 2 mRNA in AT or SM. Thus hypocaloric diet and increased physical activity improved insulin sensitivity and reduced low-grade inflammation. Markers of inflammation were particularly reduced in AT, whereas SM does not contribute to this attenuation of whole body inflammation. PMID- 16352668 TI - Effects of diabetes and insulin on betaine-homocysteine S-methyltransferase expression in rat liver. AB - Elevation of plasma homocysteine levels has been recognized as an independent risk factor for the development of cardiovascular disease, a major complication of diabetes. Plasma homocysteine reflects a balance between its synthesis via S adenosyl-L-methionine-dependent methylation reactions and its removal through the transmethylation and the transsulfuration pathways. Betaine-homocysteine methyltransferase (BHMT, EC 2.1.1.5) is one of the enzymes involved in the remethylation pathway. BHMT, a major zinc metalloenzyme in the liver, catalyzes the transfer of methyl groups from betaine to homocysteine to form dimethylglycine and methionine. We have previously shown that plasma homocysteine levels and the transsulfuration pathway are affected by diabetes. In the present study, we found increased BHMT activity and mRNA levels in livers from streptozotocin-diabetic rats. In the rat hepatoma cell line (H4IIE cells), glucocorticoids (triamcinolone) increased the level and rate of BHMT mRNA synthesis. In the same cell line, insulin decreased the abundance of BHMT mRNA and the rate of de novo mRNA transcription of the gene. Thus the decreased plasma homocysteine in various models of diabetes could be due to enhanced homocysteine removal brought about by a combination of increased transsulfuration of homocysteine to cysteine and increased remethylation of homocysteine to methionine by BHMT. PMID- 16352669 TI - Effect of medical castration on CD4+ CD25+ T cells, CD8+ T cell IFN-gamma expression, and NK cells: a physiological role for testosterone and/or its metabolites. AB - The higher prevalence of autoimmune disease among women compared with men suggests that steroids impact immune regulation. To investigate how sex steroids modulate cellular immune function, we conducted a randomized trial in 12 healthy men aged 35-55 yr treated for 28 days with placebo, a GnRH antagonist, acyline to induce medical castration, or acyline plus daily testosterone (T) gel to replace serum T, followed by a 28-day recovery period. Serum hormones were measured weekly and peripheral blood lymphocytes (PBLs) were collected biweekly for analyses of thymus-derived lymphocyte (T cell) subtypes and natural killer (NK) cells. Compared with the other groups and to baseline throughout the drug exposure period, men receiving acyline alone had significant reductions in serum T (near or below castrate levels), dihydrotestosterone, and estradiol (P < 0.05). Medical castration significantly reduced the percentage of CD4+ CD25+ T cells (P < 0.05), decreased mitogen-induced CD8+ T cell IFN-gamma expression, and increased the percentage of NK cells without affecting the ratio of CD4+ to CD8+ T cells and the expression of NK cell-activating receptor NKG2D or homing receptor CXCR1. No changes in immune composition were observed in subjects receiving placebo or acyline with replacement T. These data suggest that T and/or its metabolites may help maintain the physiological balance of autoimmunity and protective immunity by preserving the number of regulatory T cells and the activation of CD8+ T cells. In addition, sex steroids suppress NK cell proliferation. This study supports a complex physiological role for T and/or its metabolites in immune regulation. PMID- 16352670 TI - Effects of streptozotocin-induced diabetes and physical training on gene expression of extracellular matrix proteins in mouse skeletal muscle. AB - Diabetes induces changes in the structure and function of the extracellular matrix (ECM) in many tissues. We investigated the effects of diabetes, physical training, and their combination on the gene expression of ECM proteins in skeletal muscle. Mice were divided to control (C), training (T), streptozotocin induced diabetic (D), and diabetic training (DT) groups. Training groups (T, DT) performed 1, 3, or 5 wk of endurance training on a treadmill. Gene expression of calf muscles was analyzed using microarray and quantitative PCR. Training group samples were collected 24 h after the last training session. Diabetes affected the gene expression of several collagens (types I, III, IV, V, VI, and XV), some noncollagenous glycoproteins, and proteoglycans (e.g., elastin, thrombospondin-1, laminin-2, decorin). Reduced gene expression of collagens in diabetic skeletal muscle was partially attenuated as a result of physical training. In diabetes, mRNA expression of the basement membrane (BM) collagens decreased and that of noncollagenous glycoproteins increased. This may change the structure of the BM in a less collagenous direction and affect its properties. PMID- 16352671 TI - LKB1-AMPK signaling in muscle from obese insulin-resistant Zucker rats and effects of training. AB - AMPK is a key regulator of fat and carbohydrate metabolism. It has been postulated that defects in AMPK signaling could be responsible for some of the metabolic abnormalities of type 2 diabetes. In this study, we examined whether insulin-resistant obese Zucker rats have abnormalities in the AMPK pathway. We compared AMPK and ACC phosphorylation and the protein content of the upstream AMPK kinase LKB1 and the AMPK-regulated transcriptional coactivator PPARgamma coactivator-1 (PGC-1) in gastrocnemius of sedentary obese Zucker rats and sedentary lean Zucker rats. We also examined whether 7 wk of exercise training on a treadmill reversed abnormalities in the AMPK pathway in obese Zucker rats. In the obese rats, AMPK phosphorylation was reduced by 45% compared with lean rats. Protein expression of the AMPK kinase LKB1 was also reduced in the muscle from obese rats by 43%. In obese rats, phosphorylation of ACC and protein expression of PGC-1alpha, two AMPK-regulated proteins, tended to be reduced by 50 (P = 0.07) and 35% (P = 0.1), respectively. There were no differences in AMPKalpha1, alpha2, -beta1, -beta2, and -gamma3 protein content between lean and obese rats. Training caused a 1.5-fold increase in AMPKalpha1 protein content in the obese rats, although there was no effect of training on AMPK phosphorylation and the other AMPK isoforms. Furthermore, training also significantly increased LKB1 and PGC-1alpha protein content 2.8- and 2.5-fold, respectively, in the obese rats. LKB1 protein strongly correlated with hexokinase II activity (r = 0.75, P = 0.001), citrate synthase activity (r = 0.54, P = 0.02), and PGC-1alpha protein content (r = 0.81, P < 0.001). In summary, obese insulin-resistant rodents have abnormalities in the LKB1-AMPK-PGC-1 pathway in muscle, and these abnormalities can be restored by training. PMID- 16352672 TI - Regulation of BCRP/ABCG2 expression by progesterone and 17beta-estradiol in human placental BeWo cells. AB - The breast cancer resistance protein (BCRP) is abundant in the placenta and protects the fetus by limiting placental drug penetration. We hypothesize that pregnancy-specific hormones regulate BCRP expression. Hence, we examined the effects of progesterone (P4) and 17beta-estradiol (E2) on BCRP expression in the human placental BeWo cells. P4 and E2 significantly increased and decreased BCRP protein and mRNA, respectively. Likewise, treatment with P4 and E2 increased and decreased, respectively, fumitremorgin C-inhibitable mitoxantrone efflux activity of BeWo cells. Reduction in BCRP expression by E2 was abrogated by the estrogen receptor (ER) antagonist ICI-182,780. However, the progesterone receptor (PR) antagonist RU-486 had no effect on P4-mediated induction of BCRP. P4 together with E2 further increased BCRP protein and mRNA compared with P4 treatment alone. This combined effect on BCRP expression was abolished by RU-486, ICI-182,780, or both. Further analysis revealed that E2 significantly decreased ER beta mRNA and strongly induced PR(B) mRNA in a dose-dependent manner but had no effect on PR(A) and ER alpha. P4 alone had no significant effect on mRNA of ER alpha, ER beta, PR(A), and PR(B). E2 in combination with P4 increased PR(B) mRNA, but the level of induction was significantly reduced compared with E2 treatment alone. Taken together, these results indicate that E2 by itself likely downregulates BCRP expression through an ER, possibly ER beta. P4 alone upregulates BCRP expression via a mechanism other than PR. P4 in combination with E2 further increases BCRP expression, presumably via a nonclassical PR- and/or E2-mediated synthesis of PR(B). PMID- 16352673 TI - Ascorbic acid-independent synthesis of collagen in mice. AB - The mouse has become the most important model organism for the study of human physiology and disease. However, until the recent generation of mice lacking the enzyme gulanolactone oxidase (Gulo), the final enzyme in the ascorbic acid biosynthesis pathway, examination of the role of ascorbic acid in various biochemical processes using this model organism has not been possible. In the mouse, similar to most mammals but unlike humans who carry a mutant copy of this gene, Gulo produces ascorbic acid from glucose. We report here that, although ascorbic acid is essential for survival, its absence does not lead to measurable changes in proline hydroxylation. Vitamin C deficiency had no significant effect on the hydroxylation of proline and collagen production during tumor growth or in angiogenesis associated with tumor or mammary gland growth. This suggests that factors other than ascorbic acid can support proline hydroxylation and collagen synthesis in vivo. Furthermore, the failure of Gulo-/- mice to thrive on a vitamin C-deficient diet therefore suggests that ascorbic acid plays a critical role in survival other than the maintenance of the vasculature. PMID- 16352674 TI - Exercise training restores uncoupling protein-3 content in limb muscles of patients with chronic obstructive pulmonary disease. AB - Oxidative capacity and uncoupling protein-3 (UCP3) content are reduced in limb muscles of patients with chronic obstructive pulmonary disease (COPD). It has been hypothesized that the physiological role of UCP3 is to protect mitochondria against lipotoxicity in cases where fatty acid influx exceeds the capacity to oxidize them. Exercise training improves oxidative capacity and reduces UCP3 protein content in healthy subjects, but the response of UCP3 to training in COPD is unknown. We studied the effect of exercise training on UCP3 content in limb muscles of COPD patients. For this, seven healthy age-matched subjects and thirteen patients with COPD were studied. All patients were admitted to an 8-wk exercise training intervention. Exercise capacity was assessed by means of an incremental cycle ergometry test. Biopsies were taken from the vastus lateralis in which UCP3 and lipid peroxidation levels were determined by Western blotting. Citrate synthase and 3-hydroxyacyl-CoA dehydrogenase (HAD; an enzyme involved in fatty acid oxidation) were measured as indexes of muscle oxidative capacity. UCP3 in COPD was approximately 50% lower compared with healthy age-matched controls. In COPD, training induced upregulation of UCP3 [from 67.7 (SD 41.8) to 113.8 (SD 104.2) arbitrary units (AU), P = 0.062], especially in the patients who showed no increase in HAD activity [from 80.9 (SD 52.6) to 167.9 (SD 109.1) AU, P = 0.028], whereas lipid peroxidation levels remained unaltered. We conclude that exercise training can restore muscle UCP3 protein level in COPD, and the nature of this response complies with the hypothesis that UCP3 may protect against lipotoxicity. PMID- 16352675 TI - Intravenous administration of amino acids during anesthesia stimulates muscle protein synthesis and heat accumulation in the body. AB - The present study was conducted to determine the contribution of muscle protein synthesis to the prevention of anesthesia-induced hypothermia by intravenous administration of an amino acid (AA) mixture. We examined the changes of intraperitoneal temperature (Tcore) and the rates of protein synthesis (K(s)) and the phosphorylation states of translation initiation regulators and their upstream signaling components in skeletal muscle in conscious (Nor) or propofol anesthetized (Ane) rats after a 3-h intravenous administration of a balanced AA mixture or saline (Sal). Compared with Sal administration, the AA mixture administration markedly attenuated the decrease in Tcore in rats during anesthesia, whereas Tcore in the Nor-AA group became slightly elevated during treatment. Stimulation of muscle protein synthesis resulting from AA administration was observed in each case, although K(s) remained lower in the Ane AA group than in the Nor-Sal group. AA administration during anesthesia significantly increased insulin concentrations to levels approximately 6-fold greater than in the Nor-AA group and enhanced phosphorylation of eukaryotic initiation factor 4E-binding protein-1 (4E-BP1) and ribosomal protein S6 protein kinase relative to all other groups and treatments. The alterations in the Ane-AA group were accompanied by hyperphosphorylation of protein kinase B and the mammalian target of rapamycin (mTOR). These results suggest that administration of an AA mixture during anesthesia stimulates muscle protein synthesis via insulin-mTOR-dependent activation of translation initiation regulators caused by markedly elevated insulin and, thereby, facilitates thermal accumulation in the body. PMID- 16352676 TI - Cardiac metabolism in mice: tracer method developments and in vivo application revealing profound metabolic inflexibility in diabetes. AB - Studies of cardiac fuel metabolism in mice have been almost exclusively conducted ex vivo. The major aim of this study was to assess in vivo plasma FFA and glucose utilization by the hearts of healthy control (db/+) and diabetic (db/db) mice, based on cardiac uptake of (R)-2-[9,10-(3)H]bromopalmitate ([3H]R-BrP) and 2 deoxy-D-[U-14C]glucose tracers. To obtain quantitative information about the evaluation of cardiac FFA utilization with [3H]R-BrP, simultaneous comparisons of [3H]R-BrP and [14C]palmitate ([14C]P) uptake were first made in isolated perfused working hearts from db/+ mice. It was found that [3H]R-BrP uptake was closely correlated with [14C]P oxidation (r2 = 0.94, P < 0.001). Then, methods for in vivo application of [3H]R-BrP and [14C]2-DG previously developed for application in the rat were specially adapted for use in the mouse. The method yields indexes of cardiac FFA utilization (R(f)*) and clearance (K(f)*), as well as glucose utilization (R(g)'). Finally, in the main part of the study, the ability of the heart to switch between FFA and glucose fuels (metabolic flexibility) was investigated by studying anesthetized, 8-h-fasted control and db/db mice in either the basal state or during glucose infusion. In control mice, glucose infusion raised plasma levels of glucose and insulin, raised R(g)' (+58%), and lowered plasma FFA level (-48%), K(f)* (-45%), and R(f)* (-70%). This apparent reciprocal regulation of glucose and FFA utilization by control hearts illustrates metabolic flexibility for substrate use. By contrast, in the db/db mice, glucose infusion raised glucose levels with no apparent influence on cardiac FFA or glucose utilization. In conclusion, tracer methodology for assessing in vivo tissue-specific plasma FFA and glucose utilization has been adapted for use in mice and reveals a profound loss of metabolic flexibility in the diabetic db/db heart, suggesting a fixed level of FFA oxidation in fasted and glucose-infused states. PMID- 16352677 TI - Inosine released after hypoxia activates hepatic glucose liberation through A3 adenosine receptors. AB - Inosine, an endogenous nucleoside, has recently been shown to exert potent effects on the immune, neural, and cardiovascular systems. This work addresses modulation of intermediary metabolism by inosine through adenosine receptors (ARs) in isolated rat hepatocytes. We conducted an in silico search in the GenBank and complete genomic sequence databases for additional adenosine/inosine receptors and for a feasible physiological role of inosine in homeostasis. Inosine stimulated glycogenolysis (approximately 40%, EC50 4.2 x 10(-9) M), gluconeogenesis (approximately 40%, EC50 7.8 x 10(-9) M), and ureagenesis (approximately 130%, EC50 7.0 x 10(-8) M) compared with basal values; these effects were blunted by the selective A3 AR antagonist 9-chloro-2-(2-furanyl)-5 [(phenylacetyl)amino][1,2,4]-triazolo[1,5-c]quinazoline (MRS 1220) but not by selective A1, A2A, and A2B AR antagonists. In addition, MRS 1220 antagonized inosine-induced transient increase (40%) in cytosolic Ca2+ and enhanced (90%) glycogen phosphorylase activity. Inosine-induced Ca2+ mobilization was desensitized by adenosine; in a reciprocal manner, inosine desensitized adenosine action. Inosine decreased the cAMP pool in hepatocytes when A1, A2A, and A2B AR were blocked by a mixture of selective antagonists. Inosine-promoted metabolic changes were unrelated to cAMP decrease but were Ca2+ dependent because they were absent in hepatocytes incubated in EGTA- or BAPTA-AM-supplemented Ca2+-free medium. After in silico analysis, no additional cognate adenosine/inosine receptors were found in human, mouse, and rat. In both perfused rat liver and isolated hepatocytes, hypoxia/reoxygenation produced an increase in inosine, adenosine, and glucose release; these actions were quantitatively greater in perfused rat liver than in isolated cells. Moreover, all of these effects were impaired by the antagonist MRS 1220. On the basis of results obtained, known higher extracellular inosine levels under ischemic conditions, and inosine's higher sensitivity for stimulating hepatic gluconeogenesis, it is suggested that, after tissular ischemia, inosine contributes to the maintenance of homeostasis by releasing glucose from the liver through stimulation of A3 ARs. PMID- 16352678 TI - Impact of exercise training on insulin sensitivity, physical fitness, and muscle oxidative capacity in first-degree relatives of type 2 diabetic patients. AB - First-degree relatives of type 2 diabetic patients (offspring) are often characterized by insulin resistance and reduced physical fitness (VO2 max). We determined the response of healthy first-degree relatives to a standardized 10-wk exercise program compared with an age-, sex-, and body mass index-matched control group. Improvements in VO2 max (14.1 +/- 11.3 and 16.1 +/- 14.2%; both P < 0.001) and insulin sensitivity (0.6 +/- 1.4 and 1.0 +/- 2.1 mg x kg(-1) x min(-1); both P < 0.05) were comparable in offspring and control subjects. However, VO2 max and insulin sensitivity in offspring were not related at baseline as in the controls (r = 0.009, P = 0.96 vs. r = 0.67, P = 0.002). Likewise, in offspring, exercise induced changes in VO2 max did not correlate with changes in insulin sensitivity as opposed to controls (r = 0.06, P = 0.76 vs. r = 0.57, P = 0.01). Skeletal muscle oxidative capacity tended to be lower in offspring at baseline but improved equally in both offspring and controls in response to exercise training (delta citrate synthase enzyme activity 26 vs. 20%, and delta cyclooxygenase enzyme activity 25 vs. 23%. Skeletal muscle fiber morphology and capillary density were comparable between groups at baseline and did not change significantly with exercise training. In conclusion, this study shows that first degree relatives of type 2 diabetic patients respond normally to endurance exercise in terms of changes in VO2 max and insulin sensitivity. However, the lack of a correlation between the VO2 max and insulin sensitivity in the first degree relatives of type 2 diabetic patients indicates that skeletal muscle adaptations are dissociated from the improvement in VO2 max. This could indicate that, in first-degree relatives, improvement of insulin sensitivity is dissociated from muscle mitochondrial functions. PMID- 16352679 TI - Effects of selective testosterone and estradiol withdrawal on skeletal sensitivity to parathyroid hormone in men. AB - CONTEXT: Gonadal steroid withdrawal increases bone turnover and causes bone loss in men, but the underlying mechanisms have not been defined. We previously reported that gonadal steroid deprivation increases the skeletal sensitivity to the bone resorbing properties of PTH infusion in men, but it is not known whether this effect is mediated by the absence of androgens, estrogens, or both. OBJECTIVE: The objective of the study was to determine the selective effects of testosterone and estradiol withdrawal on the skeletal sensitivity to PTH infusion in healthy adult men. DESIGN AND SETTING: We randomly assigned 58 healthy men between the ages of 20 and 45 yr to receive treatment with combinations of a GnRH analog, an aromatase inhibitor, and hormone add-back therapy to produce the following treatment groups: group 1 (testosterone and estradiol deficient, n = 16); group 2 (testosterone sufficient but estradiol deficient, n = 12); group 3 (testosterone deficient but estradiol sufficient, n = 14); and group 4 (testosterone and estradiol sufficient, n = 16). Twenty-four-hour PTH infusions were performed at baseline and after 6 wk of therapy. Serum N-telopeptide (NTX), C-telopeptide (CTX), osteocalcin (OC), and amino-terminal propeptide of type I procollagen (P1NP) were measured every 6 h during the PTH infusions. RESULTS: Serum testosterone levels fell into the castrate range in groups 1 and 3, whereas estradiol levels were similarly reduced in groups 1 and 2. Gonadal steroid levels in the replaced groups were unchanged from baseline. Serum NTX levels measured before PTH infusion did not change in group 4 (+T, +E) but increased significantly in all other groups. A similar pattern was observed in serum CTX, although the increase in group 2 (+T, -E) was not significant (P = 0.12). Preinfusion concentrations of both OC and P1NP fell in most groups, but these changes were significant in group 2 (+T, -E) for both OC and P1NP and group 4 (+T, +E) for P1NP only. Serum NTX and CTX increased during PTH infusions in all groups at all time points (P < 0.001). In the eugonadal group (group 4 +T+E), the increase in NTX was the same at wk 0 and 6, whereas in all the other groups, the PTH-induced increase in serum NTX was significantly greater at wk, 6 compared with wk 0. The same pattern emerged for CTX, although the difference in group 3 ( T,+E) was not significant (P = 0.12). Serum OC and P1NP levels fell during PTH infusions in all groups and at all time points (P < 0.001), but no significant differences were observed between wk 0 and 6 in any group. CONCLUSIONS: These results demonstrate that the selective suppression of testosterone, estradiol, or both hormones increases the skeletal responsiveness to the bone-resorbing effects of PTH in men. These findings underscore the importance of both androgens and estrogens in male skeletal homeostasis and suggest that changes in skeletal sensitivity to PTH may play an important role in the pathogenesis of hypogonadal bone loss in men. PMID- 16352680 TI - Early effects of metformin in women with polycystic ovary syndrome: a prospective randomized, double-blind, placebo-controlled trial. AB - CONTEXT: Metformin is successfully used in the treatment of cycle disorders and anovulation in women with polycystic ovary syndrome (PCOS). No data of the exact point and the impact of insulin resistance (IR) on metformin's efficacy exist. OBJECTIVE: The objective of the study was to evaluate the early potential effects of metformin treatment, their time of onset, and the role of IR on metformin's efficacy. DESIGN: This was a prospective randomized, double-blind, placebo controlled trial. SETTING: The study was conducted at the University of Heidelberg, Heidelberg, Germany. PATIENTS: The patient population was 45 oligo /anovulatory PCOS women with typical ovaries. INTERVENTIONS: Women were stratified for IR (32 of 13) and then randomly allocated to receive either metformin (n = 22) or placebo (n = 23) and were assessed before and every 4 wk within a treatment period of 12 wk. MAIN OUTCOME MEASURES: Menstrual disturbance and markers of insulin metabolism were measured. RESULTS: The main outcome criterion menstrual disturbance was successfully improved in the metformin treated group, depending on IR (12 of 15 vs. three of 17), whereas women without IR (four of seven vs. four of six) had no significant amelioration of their menstrual irregularities (P < 0.05). Estradiol levels increased continuously only in the treatment group (P < 0.005), indicating an improvement of ovulatory function. Sixty-seven percent of metformin-treated women had at least one ovulation, compared with only 45% in the placebo group, shown by biphasic body temperature curves. Insulin sensitivity improved within 4 wk after beginning of metformin as shown by an increased area under the curve glucose to insulin ratio, compared with baseline (P < 0.005). CONCLUSIONS: IR is a baseline predictor of clinical efficacy in metformin treatment in PCOS women measured by improved menstrual cyclicity and ovulatory function. PMID- 16352681 TI - Prospective study of changes in bone mineral density and turnover in children after hematopoietic cell transplantation. AB - CONTEXT: Osteoporosis is common in adults after hematopoietic cell transplantation (HCT). The data on bone mineral density (BMD) in children after HCT are limited. OBJECTIVE: The objective of the study was to determine the incidence, timing, magnitude, and possible predictors of bone loss in children after HCT. PATIENTS AND DESIGN: The study population included 49 patients (age 5 18 yr) who were eligible to receive HCT at the University of Minnesota. The patients were evaluated at baseline, 100 d, 6 months, and 1 yr after HCT. Lumbar BMD (LBMD) was assessed by dual-energy x-ray absorptiometry. RESULTS: The number of patients with osteopenia increased from 18% at baseline to 33% 1 yr after HCT, and with osteoporosis from 16-19%. Mean areal LBMD z-score decreased from -0.56 to -1.1 by 6 months (n = 27) and at 1 yr was -0.94 (n = 21), which was significant compared with standard normal distribution (P = 0.004 and P = 0.022, respectively). The absolute loss of bone mineral corresponded to a 5.3% reduction in areal LBMD and a 4.8% reduction in volumetric LBMD. The level of bone-specific alkaline phosphatase decreased by 30% by d 100 (P = 0.009), followed by recovery toward baseline by 6 months. The level of osteocalcin greater than 6.5 ng/ml at d 100 predicted recovery from the initial bone loss by 1 yr. A reduction in LBMD at 6 months correlated with a cumulative dose of glucocorticoids. CONCLUSION: This study demonstrates that bone loss is common in children after HCT and is primarily due to suppression of bone formation. Further studies are necessary to validate osteocalcin as a predictive biomarker. PMID- 16352682 TI - Fibroblast growth factor 23 is increased in calcium nephrolithiasis with hypophosphatemia and renal phosphate leak. AB - CONTEXT: Nephrolithiasis affects about 10% of the population in industrialized countries, with calcium salts composing more than 80% of renal stones. A significant percentage of patients with calcium nephrolithiasis and normal parathyroid function show hypophosphatemia and reduced renal phosphate reabsorption (i.e. a renal phosphate leak). OBJECTIVES: The objective of the study was to compare serum levels of fibroblast growth factor 23 (FGF23), a regulator of phosphate homeostasis, in 110 recurrent stone formers with or without renal phosphate leak, six patients affected by X-linked hypophosphatemic rickets, five patients affected by oncogenic osteomalacia, and 60 unrelated healthy controls. DESIGN: This was a prospective interventional study. METHODS: Renal phosphate leak was identified based on the occurrence of idiopathic hypophosphatemia [serum phosphate concentration < 2.50 mg/dl (<0.80 mmol/liter)] and reduced renal threshold phosphate concentration [<2.2 mg/liter (<0.70 mmol/liter)]. RESULTS: In 22 stone formers with renal phosphate leak, serum FGF23 concentration was significantly higher as compared with 88 stone formers without renal phosphate leak and with controls [83.3 (65.6-101.1) vs. 32.1 (26.8-37.4) and 24.5 (19.8-29.1) reference units (RU)/ml, respectively]. Stone formers with renal phosphate leak showed lower FGF23, compared with patients with oncogenic osteomalacia and X-linked hypophosphatemic rickets [572.3 (235.9-908.7) RU/ml]. Among stone formers and controls, serum FGF23 concentration displayed a strong inverse association with serum phosphate (r = -0.784, P = 0.009) and the rate of tubular phosphate reabsorption (r = -0.791, P = 0.008). CONCLUSIONS: In our study population, renal phosphate leak affected 20% of stone formers and was strongly associated with increased serum FGF23 concentration. PMID- 16352683 TI - Prolonged stimulation of growth hormone (GH) and insulin-like growth factor I secretion by CJC-1295, a long-acting analog of GH-releasing hormone, in healthy adults. AB - CONTEXT: Therapeutic use of GHRH to enhance GH secretion is limited by its short duration of action. OBJECTIVE: The objective of this study was to examine the pharmacokinetic profile, pharmacodynamic effects, and safety of CJC-1295, a long acting GHRH analog. DESIGN: The study design was two randomized, placebo controlled, double-blind, ascending dose trials with durations of 28 and 49 d. SETTING: The study was performed at two investigational sites. PARTICIPANTS: Healthy subjects, ages 21-61 yr, were studied. INTERVENTIONS: CJC-1295 or placebo was administered sc in one of four ascending single doses in the first study and in two or three weekly or biweekly doses in the second study. MAIN OUTCOME MEASURES: The main outcome measures were peak concentrations and area under the curve of GH and IGF-I; standard pharmacokinetic parameters were used for CJC 1295. RESULTS: After a single injection of CJC-1295, there were dose-dependent increases in mean plasma GH concentrations by 2- to 10-fold for 6 d or more and in mean plasma IGF-I concentrations by 1.5- to 3-fold for 9-11 d. The estimated half-life of CJC-1295 was 5.8-8.1 d. After multiple CJC-1295 doses, mean IGF-I levels remained above baseline for up to 28 d. No serious adverse reactions were reported. CONCLUSIONS: Subcutaneous administration of CJC-1295 resulted in sustained, dose-dependent increases in GH and IGF-I levels in healthy adults and was safe and relatively well tolerated, particularly at doses of 30 or 60 microg/kg. There was evidence of a cumulative effect after multiple doses. These data support the potential utility of CJC-1295 as a therapeutic agent. PMID- 16352684 TI - Maternal 25-hydroxyvitamin D and parathyroid hormone concentrations and offspring birth size. AB - CONTEXT: There is inconsistent evidence that maternal 25-hydroxyvitamin D [25 (OH)D] deficiency may impair fetal growth. OBJECTIVE: The objective of the study was to examine the relationship between maternal 25-(OH)D and PTH concentrations at less than 16 and 28 wk gestation and offspring birth size. DESIGN: This was an observational study. SETTING: The study was set at a hospital antenatal clinic. PARTICIPANTS: Women with singleton pregnancies, before 16 wk gestation, participated. INTERVENTIONS: No interventions were used. MAIN OUTCOME MEASURE: Knee-heel length at birth was the main outcome measure. RESULTS: Altogether 374 of 475 (79%) women completed this study. We found no evident relationship between birth size measures and maternal 25-(OH)D or PTH at recruitment (approximately 11 wk). Gestation length was 0.7 wk (95% confidence interval -1.3, -0.1) shorter and knee-heel length was 4.3 mm smaller (-7.3, -1.3) in infants of 27 mothers with low 25-(OH)D (<28 nmol/liter) at 28-32 wk vs. babies whose mothers had higher concentrations. This latter difference was reduced to -2.7 mm (-5.4, -0.1) after adjustment for gestation length, suggesting some of the apparent growth deficit is explained by shorter gestation. There was no evidence that other birth measures were affected. Maternal PTH concentration at 28-32 wk was positively related to knee-heel length, birth weight, and mid-upper arm and calf circumferences. These associations were independent of 25-(OH)D concentration. CONCLUSIONS: Low maternal 25-(OH)D in late pregnancy is associated with reduced intrauterine long bone growth and slightly shorter gestation. The long-term consequences for linear growth and health require follow-up. The positive relationship between maternal PTH and measures of infant size may relate to increased mineral demands by larger babies, but warrants further investigation. PMID- 16352685 TI - The association of CTLA4 polymorphism with type 1 diabetes is concentrated in patients complicated with autoimmune thyroid disease: a multicenter collaborative study in Japan. AB - CONTEXT: Transracial studies are a powerful tool for genetic association studies of multifactorial diseases, such as type 1 diabetes. The low incidence of type 1 diabetes in Asian countries, however, makes it difficult to perform large-scale studies in Asia. OBJECTIVE: To overcome this, we have assembled a multicenter study group in Japan and studied the association of CTLA4 polymorphisms with type 1 diabetes relative to autoimmune thyroid disease (AITD) phenotypes. SUBJECTS: Subjects included a total of 1837 samples, including 1114 cases (769 with type 1 diabetes and 345 with AITD) and 723 control subjects. METHODS: The +6230G>A and +49G>A polymorphisms of CTLA4 as well as HLA-DRB1 and -DQB1 were genotyped. RESULTS: The +6230G>A polymorphism was significantly associated with type 1 diabetes complicated with AITD (odds ratio, 1.54; P = 0.027) and with AITD alone (odds ratio, 1.31; P = 0.045) but not with type 1 diabetes without AITD. The association with type 1 diabetes positive for autoantibodies to both pancreatic islets and thyroid was particularly strong (odds ratio, 1.87; P = 0.001). Type 1 diabetic patients with the disease-associated GG genotype were characterized by a significantly higher frequency of AITD (P = 0.013), of positivity for both AITD and antiislet autoantibody (P = 0.00086), and of high-risk HLA genotypes (P = 0.034). CONCLUSIONS: Given the high frequency of AITD in patients with type 1 diabetes, these data suggest the possibility that the association of CTLA4 with type 1 diabetes in previous studies may have been secondary to AITD, suggesting the importance of subclassification of type 1 diabetes relative to AITD in genetic studies. PMID- 16352686 TI - Influence of nutritional recovery on the leptin axis in severely malnourished children. AB - CONTEXT: Leptin might be more important as a starvation hormone than as a satiety signal. The role of the soluble leptin receptor (sOB-R) and its regulation in children with protein energy malnutrition (PEM) is poorly understood. DESIGN: We elucidated the effect of intensive nutritional support on the leptin axis in 26 severely malnourished toddlers who received infant milk-based formula for 2 wk via continuous enteral tube feeding followed by 2 wk ad libitum feeding. Serum levels of leptin, sOB-R, IGF-I, and IGF-binding protein-3 as well as anthropometric measurements were determined at the beginning of the study and at 2-wk intervals. The control group consisted of 13 well-nourished children. RESULTS: The following were changes in the PEM toddlers after the nutritional support. Leptin increased significantly (P < 0.001), reaching 166% of levels observed in control group. sOB-R decreased significantly (P < 0.001), and a 142 fold molar excess of sOB-R over leptin was found. There were significant correlations between leptin and IGF-I after 2 wk and IGF-binding protein-3 during the whole study. sOB-R was not correlated with any anthropometric data, whereas IGF-I was a predictor of sOB-R variance in the PEM toddlers (19.9%, P = 0.022). CONCLUSION: It can be concluded that sOB-R has a modulatory effect on leptin in PEM children during nutritional recovery and participates in their adaptive survival mechanisms. Leptin and the molar excess of sOB-R over leptin are better biomarkers of nutritional status than IGF-I in PEM children during nutritional recovery. PMID- 16352687 TI - The Ras effector NORE1A is suppressed in follicular thyroid carcinomas with a PAX8-PPARgamma fusion. AB - CONTEXT: The Ras effector NORE1A (RASSF5A) is a putative tumor suppressor and is inactivated in several human cancers. NORE1A has not been studied in thyroid cancer. OBJECTIVE: The objective of this study was to investigate whether NORE1A is involved in follicular thyroid cancer (FTC) development. DESIGN: We analyzed NORE1A expression in 25 FTCs, eight follicular thyroid adenomas, and seven normal thyroid tissues by TaqMan quantitative RT-PCR. The results were evaluated in relation to RASSF1A expression, RAS mutations, and PAX8-PPARgamma fusions assessed in the same material. NORE1A promoter methylation was assessed by the combined bisulfite restriction endonuclease assay. RESULTS: Although the NORE1A mRNA levels of the majority of the tumors were similar to those in the normal controls, the cases harboring a PAX8-PPARgamma translocation (n = 6) exhibited dramatically reduced NORE1A expression (P < 0.001). In contrast, RAS mutations (n = 5) and NORE1A down-regulation were mutually exclusive. A significant reduction in the expression of the NORE1A homolog and the bona fide tumor suppressor gene RASSF1A was observed, but with weak correlation to the respective NORE1A values. No NORE1A promoter methylation was detected in the 32 thyroid tumors analyzed. CONCLUSIONS: Our experiments demonstrate the suppression of NORE1A, a known Ras effector, in PAX8-PPARgamma carrying FTCs. PMID- 16352688 TI - Quinapril treatment increases insulin-stimulated endothelial function and adiponectin gene expression in patients with type 2 diabetes. AB - OBJECTIVE: Angiotensin-converting enzyme inhibitors reduce cardiovascular mortality and improve endothelial function in type 2 diabetic patients. We hypothesized that 2 months of quinapril treatment would improve insulin stimulated endothelial function and glucose uptake in type 2 diabetic subjects and simultaneously increase the expression of genes that are pertinent for endothelial function and metabolism. METHODS: Twenty-four type 2 diabetic subjects were randomized to receive 2 months of quinapril 20 mg daily or no treatment in an open parallel study. Endothelium-dependent and -independent vasodilation was studied during serotonin or sodium nitroprusside infusion in the diabetic patients and in 15 healthy subjects. Endothelial function, insulin stimulated endothelial function, and insulin-stimulated glucose uptake were measured before and after quinapril treatment. Blood flow was measured by venous occlusion plethysmography. Gene expression was measured by real-time PCR. RESULTS: Quinapril treatment increased insulin-stimulated endothelial function in the type 2 diabetic subjects (P = 0.005), whereas forearm glucose uptake was unchanged. Endothelial function was also increased by quinapril (P = 0.001). Systolic and diastolic blood pressures were reduced by quinapril (P < 0.001). Quinapril increased adiponectin gene expression in vascular tissue obtained from sc adipose biopsies. CONCLUSIONS: Quinapril treatment increases insulin stimulated endothelial function in patients with type 2 diabetes. Increased vascular adiponectin gene expression may contribute to this beneficial effect. PMID- 16352689 TI - The effect of rosiglitazone on the liver: decreased gluconeogenesis in patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: Diabetic hyperglycemia results from insulin resistance of peripheral tissues and glucose overproduction due to increased gluconeogenesis (GNG). Thiazolidinediones have been shown to improve glycemic control and increase peripheral insulin sensitivity. Whether chronic thiazolidinedione treatment is associated with a decrease in GNG has not been determined. MATERIALS AND METHODS: We studied 26 diet-treated type 2 diabetic patients randomly assigned to rosiglitazone (RSG; 8 mg/d; n = 13) or placebo (n = 13) for 12 wk. At baseline and 12 wk, we measured endogenous glucose production (by [3H]glucose infusion) and GNG (by the [2H]2O technique) after a 15-h fast. Peripheral insulin sensitivity was evaluated by a two-step (240 and 960 pmol/min/m(-2)) euglycemic insulin clamp. RESULTS: Compared with placebo, RSG reduced fasting plasma glucose (9.7 +/- 0.7 to 7.4 +/- 0.3 mmol/liter; P < 0.001), fasting fractional GNG (-15 +/- 4%; P = 0.002), and fasting GNG flux (-3.9 +/- 1.2 micromol/min/kg fat-free mass; P = 0.004), with no effect on glycogenolytic flux. Changes in GNG flux and fasting glucose were tightly correlated (r = 0.83; P < 0.0001). During both clamp steps, RSG enhanced insulin-mediated glucose clearance (by 26% and 31%; P = 0.01 and P < 0.02, respectively). In a subgroup of patients studied with magnetic resonance imaging, the reduction in GNG flux was correlated (r = 0.65; P < 0.02) with the reduction in visceral fat area. CONCLUSION/INTERPRETATION: RSG increases peripheral tissue insulin sensitivity and decreases endogenous glucose release via an inhibition of gluconeogenesis. PMID- 16352690 TI - Postprandial blood glucose is a stronger predictor of cardiovascular events than fasting blood glucose in type 2 diabetes mellitus, particularly in women: lessons from the San Luigi Gonzaga Diabetes Study. AB - OBJECTIVE: The influence of postprandial blood glucose on diabetes complications is intensively debated. We aimed to evaluate the predictive role of both fasting and postprandial blood glucose on cardiovascular events in type 2 diabetes and the influence of gender. METHODS: In a population of 529 (284 men and 245 women) consecutive type 2 diabetic patients attending our diabetes clinic, we evaluated the relationships, corrected for cardiovascular risk factors and type of treatment, between cardiovascular events in a 5-yr follow-up and baseline values of hemoglobin A1c (HbA1c) and blood glucose measured: 1) after an overnight fast, 2) after breakfast, 3) after lunch, and 4) before dinner. Continuous variables were categorized into tertiles. RESULTS: We recorded cardiovascular events in 77 subjects: 54 of 284 men (19%) and 23 of 245 women (9.4%). Univariate analysis indicated that cardiovascular events were associated with increasing age, longer diabetes duration, and higher HbA1c and fibrinogen in men, and higher systolic blood pressure, albumin excretion rate, HbA1c, and all blood glucose values in women. Smoking was more frequent in subjects with events. When all blood glucose values and HbA1c were introduced simultaneously in the models, only blood glucose after lunch predicted cardiovascular events, with hazard ratio of the third tertile vs. the first and the second tertiles greater in women (5.54; confidence interval, 1.45-21.20) than in men (2.12; confidence interval, 1.04-4.32; P < 0.01). CONCLUSIONS: Postprandial, but not fasting, blood glucose is an independent risk factor for cardiovascular events in type 2 diabetes, with a stronger predictive power in women than in men, suggesting that more attention should be paid to postprandial hyperglycemia, particularly in women. PMID- 16352691 TI - Hypothalamic and gonadal components of hypogonadism in boys with Prader-Labhart- Willi syndrome. AB - CONTEXT: The specific form of hypogonadism in Prader-Labhart-Willi syndrome (PWS), central or peripheral, remains unexplained. OBJECTIVES: The objectives of this study were to investigate the cause of hypogonadism in PWS and determine whether human chorionic gonadotropin (hCG) treatment can restore pubertal development. DESIGN: This was a clinical follow-up study, divided into two samples, over a duration of 1.5 and 4.5 yr. PATIENTS: Eight male infants and six peripubertal boys (age at start of observation, 0.06-0.93 and 8.1-10.8 yr, respectively) with genetically confirmed PWS were studied. INTERVENTION: hCG (500 1500 U twice weekly) was given from age 13.5 yr to the present. MAIN OUTCOME MEASURES: Serum FSH, LH, inhibin B, and testosterone levels and pubertal development were the main outcome measures. RESULTS: Infants with PWS presented normal LH (2.3 +/- 0.7 U/liter) and testosterone (2.5 +/- 0.9 nmol/liter) levels (mean +/- sem at 5 months) compared with the reference range. However, two thirds of the boys displayed cryptorchidism. Inhibin B levels were at the lowest level of the normal range and decreased significantly between infancy and puberty (at 13 yr, 72 +/- 17 pg/ml), whereas FSH secretion increased (9.9 +/- 2.6 U/liter). Pubertal maturation stopped at an average bone age of 13.9 yr. hCG therapy increased testosterone (11 +/- 2 nmol/liter) and reduced FSH (at 16 yr, 1.1 +/- 0.9 U/liter) levels. Testicular volume (5.6 +/- 1 ml) and inhibin B (26.5 +/- 11.9 pg/ml) remained low. CONCLUSION: Children with PWS display a specific form of combined hypothalamic (low LH) and peripheral (low inhibin B and high FSH) hypogonadism, suggesting a primary defect in Sertoli and/or germ cell maturation or an early germ cell loss. hCG therapy stimulates testosterone production and virilization. PMID- 16352692 TI - The effect of raloxifene after discontinuation of long-term alendronate treatment of postmenopausal osteoporosis. AB - OBJECTIVE: The aim of this study was to compare bone mineral density (BMD) and biochemical markers of bone turnover in patients receiving long-term alendronate therapy who continued alendronate, were switched to raloxifene, or discontinued antiresorptive therapy. DESIGN, PATIENTS, AND INTERVENTIONS: Ninety-nine ambulatory women who were diagnosed with postmenopausal osteoporosis and treated with alendronate (10 mg/d) for a mean period of 43 months were randomized to double-blind raloxifene (60 mg/d; n = 33), placebo (n = 33), or continuation of open-label alendronate (n = 33) for 12 months. Patients continued their assigned treatment in a subsequent 12-month, open-label extension phase. All patients received supplemental calcium (500 mg/d) and vitamin D (800 IU/d). MAIN OUTCOME MEASURES: BMD (lumbar spine, total femur, femoral neck, distal forearm, and total body) and biochemical markers (serum intact amino-terminal propeptide of type I procollagen, type 1 collagen cross-linked C-telopeptide, and osteocalcin) were measured at baseline and follow-up visits. RESULTS: Discontinuation of alendronate therapy resulted in a decrease in lumbar spine BMD at 12 months ( 2.66%; P < 0.05), but did not change total femur BMD (+0.35%; nonsignificant). Raloxifene and alendronate, compared with discontinuation, prevented lumbar spine BMD loss (-0.75% and -0.54% at 12 months, respectively; P < 0.05). Raloxifene and alendronate caused a similar increase in total femur BMD at 12 months (1.45% and 1.56%; both P < 0.05 vs. baseline; nonsignificant vs. discontinuation). Patients, who discontinued alendronate therapy experienced an increase in bone turnover. Bone turnover increases were less pronounced in patients taking raloxifene and were absent in those who continued alendronate. Of the three groups, mean bone turnover in raloxifene patients was the closest to premenopausal mean values. CONCLUSIONS: BMD preservation and increase were most pronounced in patients continuing alendronate. Raloxifene treatment, compared with placebo, demonstrated beneficial effects on BMD and bone turnover after discontinuation of long-term alendronate therapy. PMID- 16352693 TI - Effect of active acromegaly and its treatment on parathyroid circadian rhythmicity and parathyroid target-organ sensitivity. AB - CONTEXT: Patients with active acromegaly have increased bone turnover and skeletal abnormalities. Biochemical cure of acromegaly may represent a functional GH-deficient state and result in cortical bone loss. Reduced PTH target-organ sensitivity occurs in adult GH deficiency and may underlie the associated development of osteoporosis. OBJECTIVE: We examined the effect of active and treated acromegaly on PTH concentration and target-organ sensitivity. PATIENTS: Ten active acromegalic subjects (GH nadir > 0.3 mug/liter after 75-g oral glucose load and IGF-I above age-related reference range) and 10 matched controls participated in the study. DESIGN: Half-hourly blood and 3-h urine samples were collected on patients and controls for 24 h. Samples were analyzed for PTH, calcium (Ca), nephrogenous cAMP (NcAMP, a marker of PTH renal activity), beta C telopeptide (bone resorption marker), and procollagen type-I amino-terminal propeptide (bone formation marker). Serum calcium was adjusted for albumin (ACa). Eight acromegalic subjects who achieved biochemical cure (GH nadir < 0.3 mug/liter after 75-g oral glucose load and IGF-I within reference range) after standard surgical and/or medical treatment reattended and the protocol repeated. RESULTS: Active acromegalic subjects had higher 24-h mean PTH, NcAMP, ACa, urine Ca, beta C-telopeptide, and procollagen type I amino-terminal propeptide (P < 0.05), compared with controls. Twenty-four-hour mean PTH increased (P < 0.001) in the acromegalic subjects after treatment, whereas NcAMP and ACa decreased (P < 0.05). CONCLUSION: Increased bone turnover associated with active acromegaly may result from increased PTH concentration and action. Biochemical cure of acromegaly results in reduced PTH target-organ sensitivity indicated by increased PTH with decreased NcAMP and ACa concentrations. PTH target-organ sensitivity does not appear to return to normal after successful treatment of acromegaly in the short term and may reflect functional GH deficiency. PMID- 16352694 TI - Role of strain differences on host resistance and the transcriptional response of macrophages to infection with Yersinia enterocolitica. AB - The outcome of a host-pathogen encounter is determined by virulence factors of the pathogen and defense factors of the host. We characterized the impact of host factors [resistant (C57BL/6) or susceptible (BALB/c) genetic background and exposure to interferon (IFN)-gamma] on transcriptional responses of bone marrow derived macrophages (BMDM) to infection with Yersinia enterocolitica. IFN-gamma treatment more profoundly altered the transcriptome of BMDM than did bacterial infection or genetic background. In BALB/c BMDM, 1,161 genes were differentially expressed in response to Yersinia infection with or without IFN-gamma prestimulation. Fourteen genes (1.2%) could only be induced by BALB/c BMDM in response to Yersinia infection after IFN-gamma pretreatment. These genes inhibit apoptosis, activate NF-kappaB and Erk signaling, are chemotactic to neutrophils, and are involved in cytoskeletal reorganization, hence possibly in phagocytosis. Ten of these genes possess a common module of binding sites for Hox, Pou, and Creb transcription factors in 2 kb of upstream genomic sequence, suggesting a possible novel role of these transcription factors in regulation of immune responses. Fifty-two of one thousand fifty differentially expressed genes (4.9%) were induced more strongly by C57BL/6 BMDM in response to Yersinia infection than BALB/c BMDM. These genes activate NK cells, have antibacterial properties, or are involved in sensing chemokines and lipopolysaccharide (LPS). These data show that host resistance factors modulate a surprisingly small, but identifiable and functionally significant, portion of the macrophage transcriptome in response to Yersinia infection. PMID- 16352696 TI - BDNF-mediated enhancement of inflammation and injury in the aging heart. AB - Aging is associated with shifts in autocrine and paracrine pathways in the cardiac vasculature that may contribute to the risk of cardiovascular disease in older persons. To elucidate the molecular basis of these changes in vivo, phage display biopanning of 3- and 18-mo-old mouse hearts was performed that identified peptide epitopes with homology to brain-derived neurotrophic factor (BDNF) in old but not young phage pools. Quantification of cardiac phage binding by titration and immunostaining after injection with BDNF-like phage identified a twofold increased density of the BDNF receptor, truncated Trk B, in the aging hearts. Studies focused on the receptor ligand using a rat model of transient myocardial ischemia revealed increases in cardiac BDNF associated with local mononuclear infiltrates in 24- but not 4-mo-old rats. To investigate these changes, both 4- and 24-mo-old rat hearts were treated with intramyocardial injections of BDNF (or PBS control), demonstrating significant inflammatory increases with activated macrophage (ED1+) in BDNF-treated aging hearts compared with aging controls and similarly treated young hearts. Additional studies with permanent coronary occlusion following intramyocardial growth factor pretreatment revealed that BDNF significantly increased the extent of myocardial injury in older rat hearts (BDNF 35 +/- 10% vs. PBS 16.2 +/- 7.9% left ventricular injury; P < 0.05) without affecting younger hearts (BDNF 15 +/- 5.1% vs. PBS 14.5 +/- 6.0% left ventricular injury). Overall, these studies suggest that age-associated changes in BDNF-Trk B pathways may predispose the aging heart to increased injury after acute myocardial infarction and potentially contribute to the enhanced severity of cardiovascular disease in older individuals. PMID- 16352695 TI - Genomic analysis of neuroendocrine development of fetal brain-pituitary-adrenal axis in late gestation. AB - The present study was performed to identify the changes in genomic expression of critical components of the hypothalamus-pituitary-adrenal (HPA) axis in the second half of gestation in fetal sheep. We isolated mRNA from pituitary, hypothalamus, hippocampus, and brain stem in fetal sheep at 80, 100, 120, 130, and 145 days of gestation and 1 and 7 days after delivery (n = 4-5/group). Using real-time RT-PCR, we measured mRNA expression levels of glucocorticoid receptor (GR), mineralocorticoid receptor (MR), serum- and glucocorticoid-induced kinase-1 (sgk1), proopiomelanocortin (POMC), CRF, and arginine vasopressin (AVP). Both MR and GR were highly expressed in pituitary and hippocampus; in all tissues GR was more highly expressed than MR. AVP was more highly expressed than CRF in hypothalamus. MR, GR, and sgk1 expression were increased postnatally in brain stem, and sgk1 expression was increased postnatally in hypothalamus. GR expression was reduced in pituitary in term fetuses compared with younger ages. Hypothalamic CRF expression was increased at the end of gestation compared with younger ages, and AVP expression was increased in newborn lambs. Pituitary POMC was increased at 100 days of gestation compared with 80 days; hypothalamic POMC was increased at 120 days. Overall, the results demonstrate the expression of both MR and GR in brain regions important for control of the HPA axis. Decreases in expression of GR in pituitary at the end of gestation might contribute to the decreased corticosteroid negative feedback sensitivity at term in this species. PMID- 16352697 TI - Gibberellin regulates mitochondrial pyruvate dehydrogenase activity in rice. AB - Pyruvate dehydrogenase kinase (PDK) is a negative regulator of the mitochondrial pyruvate dehydrogenase complex (mtPDC) that plays a key role in intermediary metabolism. OsPDK1 was identified as a gibberellin-up-regulated gene using a cDNA microarray. The full-length cDNA for OsPDK1 was 1498 bp and encoded a predicted polypeptide of 363 amino acids. Genomic DNA analysis showed the presence of another isoform of PDK, OsPDK2, in rice. Reverse transcriptase-PCR analysis revealed differential expression of the two isoforms. OsPDK1 was expressed in leaf blade and leaf sheath but not in callus and root, while OsPDK2 was expressed constitutively in all tissues examined. Maximum expression of OsPDK1 in leaf sheath was detected by Northern blot analysis when seedlings were treated with 5 microM GA3 for 24 h. OsPDK1 expression was up-regulated by GA3, and there was little effect of other plant hormones. Mitochondrial pyruvate dehydrogenase (PDH) activity was reduced compared with control plants in 2-week-old seedlings treated with GA3. The beta-glucuronidase (GUS) reporter gene, driven by a 2,067 bp OsPDK1 promoter region fragment, was mainly expressed in the aleurone layer of germinating seed and leaf sheath. Transgenic rice expressing PDK1 RNAi had altered vegetative growth with reduced accumulation of vegetative tissues. These results suggest that gibberellin modulates the activity of mtPDC by regulating OsPDK1 expression and subsequently controlling plant growth and development. PMID- 16352698 TI - New evidence about the relationship between water channel activity and calcium in salinity-stressed pepper plants. AB - This study, of how Ca2+ availability (intracellular, extracellular or linked to the membrane) influences the functionality of aquaporins of pepper (Capsicum annuum L.) plants grown under salinity stress, was carried out in plants treated with NaCl (50 mM), CaCl2 (10 mM), and CaCl2 (10 mM) + NaCl (50 mM). For this, water transport through the plasma membrane of isolated protoplasts, and the involvement of aquaporins and calcium (extracellular, intracellular and linked to the membrane) has been determined. After these treatments, it could be seen that the calcium concentration was reduced in the apoplast, in the cells and on the plasma membrane of roots of pepper plants grown under saline conditions; these concentrations were increased or restored when extra calcium was added to the nutrient solution. Protoplasts extracted from plants grown under Ca2+ starvation showed no aquaporin functionality. However, for the protoplasts to which calcium was added, an increase of aquaporin functionality of the plasma membrane was observed [osmotic water permeability (Pf) inhibition after Hg addition]. Interestingly, when verapamil (a Ca2+ channel blocker) was added, no functionality was observed, even when Ca2+ was added with verapamil. Therefore, calcium seems to be involved in plasma membrane aquaporin regulation via a chain of processes within the cell but not by alteration of the stability of the plasma membrane. PMID- 16352699 TI - Metabolic transformation plays a primary role in the psychostimulant-like discriminative-stimulus effects of selegiline [(R)-(-)-deprenyl]. AB - l-Deprenyl [selegiline, (R)-(-)-deprenyl] is a selective inhibitor of monoamine oxidase B (MAO-B) used in the treatment of Parkinson's disease and proposed as an antidepressant and an aid for cigarette-smoking cessation and treatment of psychostimulant abuse. Beneficial therapeutic effects of (R)-(-)-deprenyl may also result from indirect actions. Brain levels of dopamine and beta phenylethylamine (beta-PEA), a behaviorally active endogenous trace amine, increase after (R)-(-)-deprenyl treatment due to MAO-B blockade and (R)-(-) deprenyl is metabolized to (R)-(-)-methamphetamine and (R)-(-)-amphetamine, suggesting that (R)-(-)-deprenyl may have psychostimulant-like behavioral effects. Indeed, (R)-(-)-deprenyl produces psychostimulant-like discriminative stimulus effects in experimental animals. Here, we tested the hypothesis that psychostimulant-like behavioral effects of (R)-(-)-deprenyl are mainly mediated by its metabolites. Male Fisher F344 rats were trained to discriminate i.p. injection of 1.0 mg/kg (S)-(+)-methamphetamine or 10.0 mg/kg cocaine from injection of saline using two-lever choice schedules of food delivery or stimulus shock termination. When (R)-(-)-deprenyl was tested by substitution, it had (S) (+)-methamphetamine- and cocaine-like discriminative-stimulus effects, but only at doses of 10 to 30 mg/kg, doses 10 to 20 times higher than those selective for MAO-B inhibition. Ro 16-6491 [N-(2-aminoethyl)-4-chlorobenzamide hydrochloride], a selective inhibitor of MAO-B enzyme activity without psychoactive metabolites, had no psychostimulant-like discriminative effects. In addition, blockade of (R) (-)-deprenyl's metabolism with SKF 525A (beta-DEAE-diphenylpropylacetate hydrochloride; 50 mg/kg i.p.) reduced or eliminated (R)-(-)-deprenyl's psychostimulant-like discriminative effects. When beta-PEA synthesis was blocked by NSD 1015 (m-hydroxy-benzyl-hydrazine; 30 mg/kg i.p.), there was a modest reversal of (R)-(-)-deprenyl's psychostimulant-like discriminative effects under some conditions, indicating a facilitatory modulation of the psychostimulant-like discriminative effects of (R)-(-)-deprenyl metabolites by elevated levels of beta PEA under certain conditions. PMID- 16352700 TI - Further characterization of the time-dependent vascular effects of delta9 tetrahydrocannabinol. AB - We have previously shown that over time (2 h), the active ingredient of cannabis, Delta(9)-tetrahydrocannabinol (THC), produces peroxisome proliferator-activated receptor (PPAR) gamma-mediated vasorelaxation of conduit arteries. We have now investigated whether incubation with THC affects agonist-stimulated contractile (methoxamine) and endothelium-dependent vasorelaxant (acetylcholine) responses in the rat superior mesenteric artery (G0) and aorta by myography. We have also investigated whether similar responses are observed in isolated resistance (G3) vessels of the mesenteric bed. In both the aorta and G0, incubation with 10 microM THC for 2 h, but not 10 min, significantly attenuated the contractile responses to methoxamine. This effect of THC was abolished in the presence of the enzyme catalase, which breaks down H(2)O(2), and was reduced in the presence of the superoxide dismutase inhibitor diethyldithiocarbamate (DETCA), but it was not PPARgamma-mediated. THC also inhibited calcium influx in a H(2)O(2)-dependent manner. In G0, but not the aorta, incubation with 10 muM THC for 2 h significantly enhanced endothelium-dependent vasorelaxation. This was inhibited by a PPARgamma antagonist, 2-chloro-5-nitro-N-phenylbenzamide (GW9662), catalase, and DETCA, but not by the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester. By contrast, in G3, no time-dependent vasorelaxation of precontracted arteries to THC was observed, and incubation with THC led to potentiation of contractile responses and blunting of vasorelaxation to acetylcholine, which seems to involve inhibition of endothelium-derived hyperpolarizing factor (EDHF) production, and agonist-stimulated production of EDHF. These data demonstrate further the time-dependent vascular actions of THC and also highlight the heterogenous effects of THC in different arterial types. PMID- 16352701 TI - Glycogen synthase kinase-3 phosphorylation, T-cell factor signaling activation, and cell morphology change following stimulation of thromboxane receptor alpha. AB - Previous reports showed that activation of the thromboxane receptor (TP) induced some types of cells to proliferate. We report here that TPalpha activates beta catenin/T-cell factor (Tcf)/lymphoid enhancer factor (Lef) pathway through phosphorylation of glycogen synthase kinase (GSK)-3. TP agonist [1S alpha,2alpha(Z),3beta(1E,3S),4alpha]]-7-[3-[3-hydroxy-4-(4-iodophenoxy)-1 butenyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid (I-BOP) induced both alpha and beta forms of GSK-3 phosphorylation in human embryonic kidney (HEK)293 cells stably overexpressing TPalpha (HEK293-TPalpha). N-[2-(4 Bromocinnamylamino)ethyl]-5-isoquinoline (H89), a protein kinase A (PKA) inhibitor, totally blocked the phosphorylation of GSK-3, whereas wortmannin, a phosphatidylinositol 3-kinase (PI-3 kinase) inhibitor, partially attenuated it, suggesting that PKA as well as PI-3 kinase/Akt pathway were involved in TP induced phosphorylation of GSK-3. I-BOP consistently stimulated an approximately 8-fold increase over basal Tcf/Lef reporter gene activity in HEK293-TPalpha cells. Furthermore, I-BOP-induced Tcf/Lef reporter gene activity was totally inhibited by H89 and partially inhibited by wortmannin. I-BOP also induced overexpression of Tcf/Lef downstream target gene cyclin D1. Blockade of the beta catenin expression by small interfering RNA approach attenuated I-BOP-induced expression of cyclin D1, indicating that the induction was mediated by beta catenin/Tcf/Lef pathway. Finally, I-BOP resulted in the morphology change, such as cell rounding and aggregation, in HEK293-TPalpha cells after 1-h incubation. However, HEK293-TPalpha cells were not able to revert back to normal shape even 24 h after the removal of the agonist, suggesting that the prolonged activation of the Tcf/Lef promoter induced downstream gene expression leading to cell permanent morphology change that was related to cell transformation. Together, our results showed for the first time TP agonist-induced phosphorylation of GSK-3 and activation of Tcf/Lef signaling leading to cell proliferation and transformation. PMID- 16352702 TI - Molecular mechanisms underlying rat mesenteric artery vasorelaxation induced by the nitric oxide-independent soluble guanylyl cyclase stimulators BAY 41-2272 [5 cyclopropyl-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-4 ylamine] and YC-1 [3-(5'-hydroxymethyl-2'-furyl)-1-benzyl Indazole]. AB - The aim of this study was to investigate the mechanisms of relaxation to the nitric oxide (NO)-independent soluble guanylyl cyclase (sGC) stimulators 5 cyclopropyl-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-4 ylamine (BAY 41-2272) and 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1) in the rat mesenteric artery. In endothelium-intact rings, BAY 41-2272 (0.0001-1 microM) and YC-1 (0.001-30 microM) caused concentration-dependent relaxations (pEC(50) values of 8.21 +/- 0.05 and 6.75 +/- 0.06, respectively), which were shifted to the right by 6-fold in denuded rings. The sGC inhibitor H [1,2,4]oxadiazolo [4,3,-a]quinoxalin-1-one (ODQ) (10 microM) partially attenuated the maximal responses to BAY 41-2272 and YC-1 and displaced their curves to the right by 9- to 10-fold in intact and 3-fold in denuded vessels. The NO synthesis inhibitor N(omega)-nitro-L-arginine methyl ester (100 microM) and the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (100 microM) reduced BAY 41-2272 and YC-1 relaxations, whereas the phosphodiesterase type 5 inhibitor sildenafil (0.1 microM) potentiated these responses. The phosphatase inhibitor calyculin A (50 nM) reduced the relaxant responses, and high concentrations of BAY 41-2272 (1 micorM) and YC-1 (10 microM) inhibited Ca(2+)-induced contractions in K(+)-depolarized rings. BAY 41-2272 (0.1 microM) and YC-1 (1 microM) markedly elevated cGMP levels in an ODQ-sensitive manner. Coincubation of BAY 41-2272 or YC-1 with a NO donor resulted in a synergistic inhibition of phenylephrine-induced contractions paralleled by marked increases in cGMP levels. In conclusion, BAY 41-2272 and YC-1 relax the mesenteric artery through cGMP-dependent and -independent mechanisms, including blockade of Ca(2+) influx. The synergistic responses probably reflect the direct effects of NO and NO-independent sGC stimulators on the enzyme, thus representing a potential therapeutic effect by permitting reductions of nitrovasodilator dose. PMID- 16352703 TI - 9L gliosarcoma cell proliferation and tumor growth in rats are suppressed by N hydroxy-N'-(4-butyl-2-methylphenol) formamidine (HET0016), a selective inhibitor of CYP4A. AB - The present study examined the effects of N-hydroxy-N'-(4-butyl-2 methylphenyl) formamidine (HET0016), a selective inhibitor of the formation of 20 hydroxyeicosatrienoic acid (20-HETE) on the growth of 9L rat gliosarcoma cells in vitro and in vivo. After 48 h of incubation, HET0016 reduced the proliferation of 9L in vitro by 55%, and this was associated with a fall in p42/p44 mitogen activated protein kinase and stress-activated protein kinase/c-Jun NH(2)-terminal kinase phosphorylation and increased apoptosis. HET0016 inhibited epidermal growth factor (EGF) and platelet-derived growth factor (PDGF)-induced proliferation and diminished phosphorylation of PDGF receptors. A stable 20-HETE analog increased 9L cell proliferation. In vivo, chronic administration of HET0016 (10 mg/kg/day i.p.) for 2 weeks reduced the volume of 9L tumors by 80%. This was accompanied by a 4-fold reduction in the mitotic index, a 3- to 4-fold increase in the apoptotic index, and a approximately 50% decrease in vascularization in the tumor. HET0016 treatment increased mean survival time of the animals from 17 to 22 days. Liquid chromatography/mass spectrometry experiments indicated that neither 9L cells grown in vitro nor 9L tumors removed produce 20-HETE when incubated with arachidonic acid. The normal surrounding brain tissue, however, avidly makes 20-HETE, and this activity is selectively inhibited by HET0016. These results suggest that HET0016 may be the prototype of a class of antigrowth compounds that may be efficacious for treating malignant brain tumors. In vivo, it may act in part by inhibiting the formation of 20-HETE by the surrounding tissue. However, the antiproliferative effects of HET0016 on 9L cells in vitro seem unrelated to its ability to inhibit the formation of 20 HETE. PMID- 16352704 TI - Hypocretin (orexin) receptor subtypes differentially enhance acetylcholine release and activate g protein subtypes in rat pontine reticular formation. AB - The hypothalamic peptides hypocretin-1 (orexin A) and -2 (orexin B) promote wakefulness by mechanisms that are not well understood. Defects in hypocretinergic neurotransmission underlie the human sleep disorder narcolepsy. Hypocretins alter cell excitability via two receptor subtypes, hypocretin receptor subtype 1 (hcrt-r1) and hypocretin receptor subtype 2 (hcrt-r2). This study aimed to identify G protein subtypes activated by hypocretin in rat pontine reticular nucleus oral part (PnO) and the hypocretin receptor subtype modulating acetylcholine (ACh) release in the PnO. G protein activation was quantified using in vitro [(35)S]guanylyl-5'-O-(gamma-thio)triphosphate autoradiography. ACh release was measured using in vivo microdialysis and high-performance liquid chromatography. Hypocretin-1-stimulated G protein activation was significantly decreased by pertussis toxin, demonstrating that some hypocretin receptors in rat PnO activate inhibitory G proteins. Hypocretin-1-stimulated ACh release was not blocked by pertussis toxin, supporting the conclusion that the hypocretin receptors modulating ACh release in rat PnO activate stimulatory G proteins. Hypocretin-1 and -2 each caused a concentration-dependent increase in ACh release with similar potencies, indicating that hcrt-r2 modulates ACh release in PnO. Hypocretin-1 caused a significantly greater increase in ACh release than hypocretin-2, suggesting a role for hcrt-r1 in the modulation of PnO ACh release. Taken together, these data provide the first evidence that hypocretin receptors in rat PnO signal via inhibitory and stimulatory G proteins and that ACh release in rat PnO is modulated by hcrt-r2 and hcrt-r1. One mechanism by which hypocretin promotes arousal may be to increase ACh release in the pontine reticular formation. PMID- 16352705 TI - Anti-inflammatory activity in vitro and in vivo of the protein farnesyltransferase inhibitor tipifarnib. AB - Protein farnesyltransferase inhibitors (FTIs) have shown clinical responses in hematologic malignancies, but the mechanisms are unclear. To better understand potential mechanisms of action, we have studied effects of the FTI tipifarnib on inflammatory responses in vitro and in vivo. In a human leukemia cell line THP-1, tipifarnib inhibited lipopolysaccharide (LPS)-induced transcription of chemokines [monocyte chemotactic protein (MCP)-1 and MCP-2], cytokines [interleukin (IL) 1beta, IL-6, and interferon (IFN)beta], signaling molecules (MyD88 and STAT-1), proteases [matrix metalloproteinase (MMP-9)], and receptors (urokinase receptor). Tipifarnib also inhibited LPS-induced secretion of MMP-9, IL-6, MCP-1, and IL 1beta in THP-1 cells. In primary human peripheral blood mononuclear cells, dose dependent inhibition of LPS-induced tumor necrosis factor (TNF)-alpha, IL-6, MCP 1, and IL-1beta by tipifarnib was observed with no evidence of cytotoxicity. Similar results were obtained in vivo in a murine model of LPS-induced inflammation, where pretreatment with tipifarnib resulted in significant inhibition of TNF-alpha, IL-6, MCP-1, IL-1beta, and MIP-1alpha production. Tipifarnib had no effect in vitro or in vivo on LPS-induced IL-8. Studies in THP 1 cells to address potential mechanism(s) showed that tipifarnib partially inhibited LPS-induced p38 phosphorylation. Tipifarnib significantly inhibited inhibitory subunit of nuclear factor-kappaB (NF-kappaB) (IkappaB)-alpha degradation and p65 nuclear translocation induced by LPS, but not by tumor necrosis factor-alpha, IL-1alpha, or toll-like receptor (TLR)2 ligand, suggesting that the target for inhibition of NF-kappaB activation was exclusive to the LPS/TLR4 signal pathway. The extent of IkappaB-alpha degradation inhibition did not correlate with inhibition of Ras farnesylation, indicating that Ras was not the target for the observed anti-inflammatory activity of tipifarnib. Our findings differ from those for other FTIs, which may have relevance for their dissimilar activity in specific tumor repertoires. PMID- 16352706 TI - Fatty acid amide hydrolase (-/-) mice exhibit an increased sensitivity to the disruptive effects of anandamide or oleamide in a working memory water maze task. AB - Although recent evidence suggests that fatty acid amide hydrolase (FAAH) may represent a potential therapeutic target, few published studies have investigated FAAH or its fatty acid amide substrates (FAAs) in animal models of learning and memory. Therefore, our primary goal was to determine whether FAAH (-/-) mice, which possess elevated levels of anandamide and other FAAs, would display altered performance in four Morris water maze tasks: acquisition of a hidden fixed platform, reversal learning, working memory, and probe trials. FAAH (-/-) mice failed to exhibit deficits in any task; in fact, they initially acquired the working memory task more rapidly than FAAH (+/+) mice. The second goal of this study was to investigate whether the FAAH inhibitor OL-135 (1-oxo-1[5-(2-pyridyl) 2-yl]-7-phenylheptane), anandamide, other FAAs, and methanandamide would affect working memory in both genotypes. FAAH (-/-), but not (+/+), mice displayed working memory impairments following exogenous administration of anandamide (ED(50) = 6 mg/kg) or oleamide (50 mg/kg). However, the central cannabinoid receptor (CB(1)) receptor antagonist SR141716 [N-(piperidin-1-yl)-5-(4 chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide HCl] only blocked the disruptive effects of anandamide. Methanandamide, which is not metabolized by FAAH, disrupted working memory performance in both genotypes (ED(50) = 10 mg/kg), suggesting that CB(1) receptor signaling is unaltered by FAAH deletion. In contrast, OL-135 and other FAAs failed to affect working memory in either genotype. These results suggest that FAAH deletion does not impair spatial learning but may enhance acquisition under certain conditions. More generally, FAAH may represent a novel therapeutic target that circumvents the undesirable cognitive side effects commonly associated with direct-acting cannabinoid agonists. PMID- 16352707 TI - Do subtype-selective gamma-aminobutyric acid A receptor modulators have a reduced propensity to induce physical dependence in mice? AB - Recent evidence suggests that GABA(A) receptors containing an alpha1 subunit mediate the sedative effect of diazepam, whereas receptors with an alpha2 subunit mediate this benzodiazepine's anxiolytic effect. Thus, compounds selective for GABA(A)-alpha2 receptors may offer advantages, i.e., lack of sedation, over current benzodiazepines. Whether such compounds would offer additional advantages over benzodiazepines is unclear. Here, we address the issue of physical dependence by comparing the GABA(A)-alpha1 affinity-selective drug zolpidem, the novel compounds 7-(1,1-dimethylethyl)-6-(2-methyl-2H-1,2,4-triazol-3-ylmethoxy)-3 (2,5-difluorophenyl)-1,2,4-triazolo[4,3-b]pyridazine (L-838,417) and 6-fluoro-9 methyl-2-phenyl-4-(pyrrolidin-1-yl-carbonyl)-2,9-dihydro-1H-pyridol[3,4-b]indol-1 one (SL651498) with functional selectivity for certain non-alpha(1) GABA(A) receptors, nonselective partial agonists [bretazenil, 1-[1-[3-(3 pyridyl)phenyl]benzimidazol-5-yl]ethanone O-ethyloxime (NS2710), and 5-furan-3-yl 1-(3-imidazol-1-phenyl)-1H-benzoimidazole (NS2664)], and nonselective full efficacy benzodiazepines, in a rapid precipitated withdrawal assay using the inverse agonist N-methyl-beta-carboline-3-carboxamide (FG-7142). For all compounds, we determined in vitro IC50 values to displace [3H]flunitrazepam from rat cortex and in vivo ED50 values for displacement of [3H]flunitrazepam from mouse forebrain (including length of in vivo occupancy). In the precipitated withdrawal model, compounds were administered at a dose giving approximately 80% receptor occupancy, obviating major differences in central nervous system bioavailability. Mice were administered compounds twice daily for 4 days and on day 5, 20 h after the final dose, given a dose of FG-7142 (40 mg/kg i.p.) that did not induce seizures in control animals. In mice treated with the three subtype-selective compounds, FG-7142 did not induce seizures. Moreover, there was a low propensity for FG-7142 to induce seizures in animals treated with the partial agonists, whereas seizures were clearly seen in animals treated with most benzodiazepines. Nonetheless, differences among the benzodiazepines themselves, similarities between the partial agonists and subtype-selective compounds, the in vitro/in vivo potency, and in vivo receptor exposure time data suggest a complex interaction among selectivity, efficacy, potency, and receptor exposure in determining physical dependence liability of benzodiazepine site modulators in mice. PMID- 16352708 TI - The response of Spartium junceum roots to slope: anchorage and gene factors. AB - BACKGROUND AND AIMS: Plant anchorage is governed by complex, finely regulated mechanisms that occur at a morphological, architectural and anatomical level. Spanish broom (Spartium junceum) is a woody plant frequently found on slopes--a condition that affects plant anchorage. This plant grows throughout the Mediterranean area where it plays an important role in preventing landslides. Spanish broom seedlings respond promptly to slope by altering stem and root morphology. The aim of this study was to investigate the mechanisms whereby the root system of Spanish broom seedlings adapts to ensure anchorage to the ground. METHODS: Seedlings were grown in tilted and untilted pots under controlled conditions. The root apparatus was removed at different times of growth and subjected to morphological, biomechanical and molecular analyses. KEY RESULTS: In slope-grown seedlings, changes in root system morphology, pulling strength and chemical lignin content, all features related to plant anchorage in the soil, were related to seedling age. cDNA-AFLP analysis revealed changes in the expression of several genes in root systems of slope-grown plants. BLAST analysis showed that some differentially expressed genes are homologues of genes induced by environmental stresses in other plant species, and/or are involved in the production of strengthening materials. CONCLUSION: Plants use various mechanisms/strategies to respond to slope depending on their developmental stage. PMID- 16352709 TI - Antidepressant-like activity and modulation of brain monoaminergic transmission by blockade of anandamide hydrolysis. AB - Although anecdotal reports suggest that cannabis may be used to alleviate symptoms of depression, the psychotropic effects and abuse liability of this drug prevent its therapeutic application. The active constituent of cannabis, delta9 tetrahydrocannabinol, acts by binding to brain CB1 cannabinoid receptors, but an alternative approach might be to develop agents that amplify the actions of endogenous cannabinoids by blocking their deactivation. Here, we show that URB597, a selective inhibitor of the enzyme fatty-acid amide hydrolase, which catalyzes the intracellular hydrolysis of the endocannabinoid anandamide, exerts potent antidepressant-like effects in the mouse tail-suspension test and the rat forced-swim test. Moreover, URB597 increases firing activity of serotonergic neurons in the dorsal raphe nucleus and noradrenergic neurons in the nucleus locus ceruleus. These actions are prevented by the CB1 antagonist rimonabant, are accompanied by increased brain anandamide levels, and are maintained upon repeated URB597 administration. Unlike direct CB1 agonists, URB597 does not exert rewarding effects in the conditioned place preference test or produce generalization to the discriminative effects of delta9-tetrahydrocannabinol in rats. The findings support a role for anandamide in mood regulation and point to fatty-acid amide hydrolase as a previously uncharacterized target for antidepressant drugs. PMID- 16352710 TI - Diffusion and scaling during early embryonic pattern formation. AB - Development of spatial patterns in multicellular organisms depends on gradients in the concentration of signaling molecules that control gene expression. In the Drosophila embryo, Bicoid (Bcd) morphogen controls cell fate along 70% of the anteroposterior axis but is translated from mRNA localized at the anterior pole. Gradients of Bcd and other morphogens are thought to arise through diffusion, but this basic assumption has never been rigorously tested in living embryos. Furthermore, because diffusion sets a relationship between length and time scales, it is hard to see how patterns of gene expression established by diffusion would scale proportionately as egg size changes during evolution. Here, we show that the motion of inert molecules through the embryo is well described by the diffusion equation on the relevant length and time scales, and that effective diffusion constants are essentially the same in closely related dipteran species with embryos of very different size. Nonetheless, patterns of gene expression in these different species scale with egg length. We show that this scaling can be traced back to scaling of the Bcd gradient itself. Our results, together with constraints imposed by the time scales of development, suggest that the mechanism for scaling is a species-specific adaptation of the Bcd lifetime. PMID- 16352711 TI - A mouse atlas of gene expression: large-scale digital gene-expression profiles from precisely defined developing C57BL/6J mouse tissues and cells. AB - We analyzed 8.55 million LongSAGE tags generated from 72 libraries. Each LongSAGE library was prepared from a different mouse tissue. Analysis of the data revealed extensive overlap with existing gene data sets and evidence for the existence of approximately 24,000 previously undescribed genomic loci. The visual cortex, pancreas, mammary gland, preimplantation embryo, and placenta contain the largest number of differentially expressed transcripts, 25% of which are previously undescribed loci. PMID- 16352712 TI - Crystal structure of human T cell leukemia virus protease, a novel target for anticancer drug design. AB - The successful development of a number of HIV-1 protease (PR) inhibitors for the treatment of AIDS has validated the utilization of retroviral PRs as drug targets and necessitated their detailed structural study. Here we report the structure of a complex of human T cell leukemia virus type 1 (HTLV-1) PR with a substrate based inhibitor bound in subsites P5 through P5'. Although HTLV-1 PR exhibits an overall fold similar to other retroviral PRs, significant structural differences are present in several loop areas, which include the functionally important flaps, previously considered to be structurally highly conserved. Potential key residues responsible for the resistance of HTLV-1 PR to anti-HIV drugs are identified. We expect that the knowledge accumulated during the development of anti-HIV drugs, particularly in overcoming drug resistance, will help in designing a novel class of antileukemia drugs targeting HTLV-1 PR and in predicting their drug-resistance profile. The structure presented here can be used as a starting point for the development of such anticancer therapies. PMID- 16352713 TI - Repression of beta-catenin function in malignant cells by nonsteroidal antiinflammatory drugs. AB - Activation of the Wnt/beta-catenin pathway promotes the development of several cancers and is an attractive target for chemopreventive and chemotherapeutic agents. Nonsteroidal antiinflammatory drugs (NSAIDs) have been reported to antagonize beta-catenin function, but their mechanism of action is not known. We demonstrate here that interference with beta-catenin function by NSAIDs does not correlate with cyclooxygenase (COX) inhibition. Instead, NSAID inhibition of beta catenin requires the high level expression of peroxisome proliferator-activated receptor gamma (PPAR-gamma) and its co-receptor retinoid-X-receptor alpha (RXR alpha). Immunoprecipitation experiments show that beta-catenin interacts with RXR alpha and PPAR-gamma in some malignant cells. Repression of beta-catenin dependent transcription by NSAIDs is thus indirect and depends on the coexpression of other nuclear receptors. PMID- 16352714 TI - Development of functional human embryonic stem cell-derived neurons in mouse brain. AB - Human embryonic stem cells are pluripotent entities, theoretically capable of generating a whole-body spectrum of distinct cell types. However, differentiation of these cells has been observed only in culture or during teratoma formation. Our results show that human embryonic stem cells implanted in the brain ventricles of embryonic mice can differentiate into functional neural lineages and generate mature, active human neurons that successfully integrate into the adult mouse forebrain. Moreover, this study reveals the conservation and recognition of common signals for neural differentiation throughout mammalian evolution. The chimeric model will permit the study of human neural development in a live environment, paving the way for the generation of new models of human neurodegenerative and psychiatric diseases. The model also has the potential to speed up the screening process for therapeutic drugs. PMID- 16352715 TI - Two types of precursor cells in a multipotential hematopoietic cell line. AB - The biochemistry of early stages of hematopoietic differentiation is difficult to study because only relatively small numbers of precursor cells are available. The murine EML cell line is a multipotential cell line that can be used to model some of these steps. We found that the lineage- EML precursor cells can be separated into two populations based on cell surface markers including CD34. Both populations contain similar levels of stem cell factor (SCF) receptor (c-Kit) but only the CD34+ population shows a growth response when treated with SCF. Conversely, the CD34- population will grow in the presence of the cytokine IL-3. The human beta-globin locus control region hypersensitive site 2 plays different roles on beta-globin transcription in the CD34+ and CD34- populations. The two populations are present in about equal amounts in culture, and the CD34+ population rapidly regenerates the mixed population when grown in the presence of SCF. We suggest that this system may mimic a normal developmental transition in hematopoiesis. PMID- 16352716 TI - An Asian origin for a 10,000-year-old domesticated plant in the Americas. AB - New genetic and archaeological approaches have substantially improved our understanding of the transition to agriculture, a major turning point in human history that began 10,000-5,000 years ago with the independent domestication of plants and animals in eight world regions. In the Americas, however, understanding the initial domestication of New World species has long been complicated by the early presence of an African enigma, the bottle gourd (Lagenaria siceraria). Indigenous to Africa, it reached East Asia by 9,000-8,000 before present (B.P.) and had a broad New World distribution by 8,000 B.P. Here we integrate genetic and archaeological approaches to address a set of long standing core questions regarding the introduction of the bottle gourd into the Americas. Did it reach the New World directly from Africa or through Asia? Was it transported by humans or ocean currents? Was it wild or domesticated upon arrival? Fruit rind thickness values and accelerator mass spectrometer radiocarbon dating of archaeological specimens indicate that the bottle gourd was present in the Americas as a domesticated plant by 10,000 B.P., placing it among the earliest domesticates in the New World. Ancient DNA sequence analysis of archaeological bottle gourd specimens and comparison with modern Asian and African landraces identify Asia as the source of its introduction. We suggest that the bottle gourd and the dog, two "utility" species, were domesticated long before any food crops or livestock species, and that both were brought to the Americas by Paleoindian populations as they colonized the New World. PMID- 16352717 TI - Unconscious fear influences emotional awareness of faces and voices. AB - Nonconscious recognition of facial expressions opens an intriguing possibility that two emotions can be present together in one brain with unconsciously and consciously perceived inputs interacting. We investigated this interaction in three experiments by using a hemianope patient with residual nonconscious vision. During simultaneous presentation of facial expressions to the intact and the blind field, we measured interactions between conscious and nonconsciously recognized images. Fear-specific congruence effects were expressed as enhanced neuronal activity in fusiform gyrus, amygdala, and pulvinar. Nonconscious facial expressions also influenced processing of consciously recognized emotional voices. Emotional congruency between visual and an auditory input enhances activity in amygdala and superior colliculus for blind, relative to intact, field presentation of faces. Our findings indicate that recognition of fear is mandatory and independent of awareness. Most importantly, unconscious fear recognition remains robust even in the light of a concurrent incongruent happy facial expression or an emotional voice of which the observer is aware. PMID- 16352718 TI - Augmenting neurotransmitter release by enhancing the apparent Ca2+ affinity of synaptotagmin 1. AB - Synaptotagmin 1 likely acts as a Ca2+ sensor in neurotransmitter release by Ca2+ binding to its two C2 domains. This notion was strongly supported by the observation that a mutation in the C2A domain causes parallel decreases in the apparent Ca2+ affinity of synaptotagmin 1 and in the Ca2+ sensitivity of release. However, this study was based on a single loss-of-function mutation. We now show that tryptophan substitutions in the synaptotagmin 1 C2 domains act as gain-of function mutations to increase the apparent Ca2+ affinity of synaptotagmin 1. The same substitutions, when introduced into synaptotagmin 1 expressed in neurons, enhance the Ca2+ sensitivity of release. Mutations in the two C2 domains lead to comparable and additive effects in release. Our results thus show that the apparent Ca2+ sensitivity of release is dictated by the apparent Ca2+ affinity of synaptotagmin 1 in both directions, and that Ca2+ binding to both C2 domains contributes to Ca2+ triggering of release. PMID- 16352719 TI - Leucine-rich repeat kinase 2 (LRRK2) interacts with parkin, and mutant LRRK2 induces neuronal degeneration. AB - Parkinson's disease (PD) is a disorder of movement, cognition, and emotion, and it is characterized pathologically by neuronal degeneration with Lewy bodies, which are cytoplasmic inclusion bodies containing deposits of aggregated proteins. Most PD cases appear to be sporadic, but genetic forms of the disease, caused by mutations in alpha-synuclein, parkin, and other genes, have helped elucidate pathogenesis. Mutations in leucine-rich repeat kinase 2 (LRRK2) cause autosomal-dominant Parkinsonism with clinical features of PD and with pleomorphic pathology including deposits of aggregated protein. To study expression and interactions of LRRK2, we synthesized cDNAs and generated expression constructs coding for human WT and mutant LRRK2 proteins. Expression of full-length LRRK2 in cells in culture suggests that the protein is predominately cytoplasmic, as is endogenous protein by subcellular fractionation. Using coimmunoprecipitation, we find that LRRK2, expressed in cells in culture, interacts with parkin but not with alpha-synuclein, DJ-1, or tau. A small proportion of the cells overexpressing LRRK2 contain protein aggregates, and this proportion is greatly increased by coexpression of parkin. In addition, parkin increases ubiquitination of aggregated protein. Also, mutant LRRK2 causes neuronal degeneration in both SH SY5Y cells and primary neurons. This cell model may be useful for studies of PD cellular pathogenesis and therapeutics. These findings suggest a gain-of-function mechanism in the pathogenesis of LRRK2-linked PD and suggest that LRRK2 may be involved in a pathogenic pathway with other PD-related proteins such as parkin, which may help illuminate both familial and sporadic PD. PMID- 16352720 TI - A regulator of nutritional copper signaling in Chlamydomonas is an SBP domain protein that recognizes the GTAC core of copper response element. AB - The CRR1 (Copper Response Regulator) locus, required for both activating and repressing target genes of a copper- and hypoxia-sensing pathway in Chlamydomonas, encodes a 1,232-residue candidate transcription factor with a plant-specific DNA-binding domain named SBP, ankyrin repeats, and a C-terminal Cys-rich region, with similarity to a Drosophila metallothionein. The recombinant SBP domain of Crr1 shows zinc-dependent binding to functionally defined copper response elements associated with the CYC6 and CPX1 promoters that contain a critical GTAC core sequence. Competition experiments indicate equivalent selectivity for copper-response elements from either promoter and 10-fold greater selectivity for the wild-type sequence vs. a sequence carrying a single mutation in the GTAC core. The SBP domain of Chlamydomonas Crr1 binds also to a related GTAC-containing sequence in the Arabidopsis AP1 promoter that is the binding site of a defining member of the SBP family of DNA-binding proteins. Chlamydomonas Crr1 is most similar to a subset of the Arabidopsis SBP domain proteins, which include SPL1, SPL7, and SPL12. The abundance of the CRR1 mRNA is only marginally copper-responsive, and although two mRNAs that differ with respect to splicing of the first intron are detected, there is no indication that the splicing event is regulated by metal nutrition or hypoxia. It is likely that the dramatic copper responsive action of Crr1 occurs at the level of the polypeptide. PMID- 16352721 TI - Information-based clustering. AB - In an age of increasingly large data sets, investigators in many different disciplines have turned to clustering as a tool for data analysis and exploration. Existing clustering methods, however, typically depend on several nontrivial assumptions about the structure of data. Here, we reformulate the clustering problem from an information theoretic perspective that avoids many of these assumptions. In particular, our formulation obviates the need for defining a cluster "prototype," does not require an a priori similarity metric, is invariant to changes in the representation of the data, and naturally captures nonlinear relations. We apply this approach to different domains and find that it consistently produces clusters that are more coherent than those extracted by existing algorithms. Finally, our approach provides a way of clustering based on collective notions of similarity rather than the traditional pairwise measures. PMID- 16352723 TI - Profile of Raymond C. Kelly. PMID- 16352722 TI - Interrogating multiple aspects of variation in a full resequencing data set to infer human population size changes. AB - We present an expanded data set of 50 unlinked autosomal noncoding regions, resequenced in samples of Hausa from Cameroon, Italians, and Chinese. We use these data to make inferences about human demographic history by using a technique that combines multiple aspects of genetic data, including levels of polymorphism, the allele frequency spectrum, and linkage disequilibrium. We explore an extensive range of demographic parameters and demonstrate that our method of combining multiple aspects of the data results in a significant reduction of the compatible parameter space. In agreement with previous reports, we find that the Hausa data are compatible with demographic equilibrium as well as a set of recent population expansion models. In contrast to the Hausa, when multiple aspects of the data are considered jointly, the non-Africans depart from an equilibrium model of constant population size and are compatible with a range of simple bottleneck models, including a 50-90% reduction in effective population size occurring some time after the appearance of modern humans in Africa 160,000 120,000 years ago. PMID- 16352724 TI - Generating highly ordered DNA nanostrand arrays. AB - Highly ordered arrays of stretched DNA molecules were generated over the millimeter scale by using a modified molecular combing method and soft lithography. Topological micropatterning on polydimethyl siloxane stamps was used to mediate the dynamic assembly of DNA molecules into arranged nonostrand arrays. These arrays consisted of either short nanostrands of several micrometers with fixed length and orientation or long nanostrands up to several hundred micrometers in length. The nanostrand arrays were transferred onto flat solid surfaces by contact printing, allowing for the creation of more complex patterns. This technique has potential applications for the construction of next-generation DNA chips and functional circuits of DNA-based 1D nanostructures. PMID- 16352725 TI - Galactocerebrosidase-deficient oligodendrocytes maintain stable central myelin by exogenous replacement of the missing enzyme in mice. AB - Globoid cell leukodystrophy (GLD) is a lysosomal storage disease caused by genetic deficiency of galactocerebrosidase (GALC) activity. Failure in catalyzing the degradation of its major substrate, galactocerebroside, in oligodendrocytes (OLs) and Schwann cells leads to death of these myelinating cells, progressive demyelination, and early demise of GLD patients. Transplantation of bone marrow cells and umbilical cord blood have been attempted as a means of enzyme replacement and have shown limited success. It remains unknown whether or how these therapies support survival of GALC-deficient OLs and myelin maintenance. We report that, upon transplantation, GALC-deficient OLs from the twitcher mouse, a model of GLD, achieved widespread myelination in the brain and spinal cord of the myelin-deficient shiverer mouse, which was preserved for the life of the host. GALC immunohistochemistry showed direct evidence for GALC transfer from the shiverer environment to the engrafted mutant OLs in vivo. These findings suggest that the mutant OLs can internalize exogenous GALC and maintain stable myelin, demonstrating that exogenous enzyme replacement will be a key strategy in the therapy of GLD. PMID- 16352726 TI - Controlling ligand substitution reactions of organometallic complexes: tuning cancer cell cytotoxicity. AB - Organometallic compounds offer broad scope for the design of therapeutic agents, but this avenue has yet to be widely explored. A key concept in the design of anticancer complexes is optimization of chemical reactivity to allow facile attack on the target site (e.g., DNA) yet avoid attack on other sites associated with unwanted side effects. Here, we consider how this result can be achieved for monofunctional "piano-stool" ruthenium(II) arene complexes of the type [(eta6 arene)Ru(ethylenediamine)(X)]n+. A potentially important activation mechanism for reactions with biomolecules is hydrolysis. Density functional calculations suggested that aquation (substitution of X by H2O) occurs by means of a concerted ligand interchange mechanism. We studied the kinetics and equilibria for hydrolysis of 21 complexes, containing, as X, halides and pseudohalides, pyridine (py) derivatives, and a thiolate, together with benzene (bz) or a substituted bz as arene, using UV-visible spectroscopy, HPLC, and electrospray MS. The x-ray structures of six complexes are reported. In general, complexes that hydrolyze either rapidly {e.g., X = halide [arene = hexamethylbenzene (hmb)]} or moderately slowly [e.g., X = azide, dichloropyridine (arene = hmb)] are active toward A2780 human ovarian cancer cells, whereas complexes that do not aquate (e.g., X = py) are inactive. An intriguing exception is the X = thiophenolate complex, which undergoes little hydrolysis and appears to be activated by a different mechanism. The ability to tune the chemical reactivity of this class of organometallic ruthenium arene compounds should be useful in optimizing their design as anticancer agents. PMID- 16352727 TI - Modulation of the electronic structure and the Ni-Fe distance in heterobimetallic models for the active site in [NiFe]hydrogenase. AB - Reaction of the mononuclear Ni(II) thiolate complexes [Ni(L)] [L, L(1), H2L(1), bis(2-mercaptoethyl)-1,2-dimercaptoethane; L(2), H2L(2), N,N'-dimethyl-N,N'-bis(2 mercaptoethyl)-bis(aminoethyl)sulfide] with [FeCp(CO)2I] gives the dithiolate bridged heterobimetallic species, [Ni(L(1))FeCp(CO)]PF6, 1, and [Ni(L(2))FeCp]I, 2, respectively. Binding of a Fe(CO)3 fragment via reaction of square-planar [Ni(pdt)(dppe)] (dppe, 1,2-diphenylphosphinoethane; pdt(2-), 1,3 propanedithiolate) with Fe3(CO)12 or [Fe(CO)3(BDA)] (BDA, benzylidene acetone) affords diamagnetic [(dppe)Ni(mu-pdt)Fe(CO)3], 3, in which the Ni(II) center is bound tetrahedrally to two thiolate S-donors and to two P-donors. The complex [(dppe)Ni(mu-pdt)Fe(CO)3], 3, reacts in solution via rearrangement to afford [(OC)Ni(mu-dppe)(mu-pdt)Fe(CO)2], 4, in which one P-donor of dppe is bound to Ni and the other to Fe, and a CO ligand has transferred from Fe to Ni. Additionally, the syntheses of 3 and 4 afford the side products [(dppe)Ni(CO)2] and [(OC)3Fe(pdt)Fe(CO)3] together with the trinuclear species [(dppe)(CO)Fe(mu CO)(mu-pdt)Fe(mu-pdt)Fe(CO)3], 5. Reaction of [Ni(pdt)(dppe)] with [FeCp(CO)2I] in CH2Cl2 affords two products [(dppe)Ni(mu-pdt)FeCp(CO)]PF6, 6, and [(dppe)Ni(pdt)(mu-I)Ni(dppe)]PF6, 7. The complexes 2, 3, and 4 show Ni-Fe distances of 2.539(4), 2.4666(6), and 2.4777(7) A, respectively, with relatively acute dihedral angles of 79.5-81.8 degrees for the Ni-S2-Fe bridge, thus mimicking the shortened Ni...Fe distance (2.5 A) and the acute dihedral angle of the Ni-S2-Fe moiety observed in certain active forms of [NiFe]hydrogenase. The role of direct Ni-Fe bonding in these complexes is discussed and linked to electronic structure calculations on [(dppe)Ni(pdt)Fe(CO)3], 3, which confirm the presence of a bent Ni(d(z2))-Fe(d(z2)) sigma-bond in a singlet ground state. PMID- 16352728 TI - Control over brain activation and pain learned by using real-time functional MRI. AB - If an individual can learn to directly control activation of localized regions within the brain, this approach might provide control over the neurophysiological mechanisms that mediate behavior and cognition and could potentially provide a different route for treating disease. Control over the endogenous pain modulatory system is a particularly important target because it could enable a unique mechanism for clinical control over pain. Here, we found that by using real-time functional MRI (rtfMRI) to guide training, subjects were able to learn to control activation in the rostral anterior cingulate cortex (rACC), a region putatively involved in pain perception and regulation. When subjects deliberately induced increases or decreases in rACC fMRI activation, there was a corresponding change in the perception of pain caused by an applied noxious thermal stimulus. Control experiments demonstrated that this effect was not observed after similar training conducted without rtfMRI information, or using rtfMRI information derived from a different brain region, or sham rtfMRI information derived previously from a different subject. Chronic pain patients were also trained to control activation in rACC and reported decreases in the ongoing level of chronic pain after training. These findings show that individuals can gain voluntary control over activation in a specific brain region given appropriate training, that voluntary control over activation in rACC leads to control over pain perception, and that these effects were powerful enough to impact severe, chronic clinical pain. PMID- 16352729 TI - Heterotrimeric G proteins precouple with G protein-coupled receptors in living cells. AB - Using fluorescence resonance energy transfer (FRET) microscopy, we investigate how heterotrimeric G proteins interact with G protein-coupled receptors (GPCRs). In the absence of receptor activation, the alpha2A adrenergic and muscarinic M4 receptors are present on the cell membrane as dimers. Furthermore, there is an interaction between the G protein subunits alpha o, beta1, and gamma2 and a number of GPCRs including M4, alpha2A, the adenosine A1 receptor, and the dopamine D2 receptor under resting conditions. The interaction between GPCRs and Galpha proteins shows specificity: there is interaction between the alpha2A receptor and Go, but little interaction between the alpha2A receptor and Gs. In contrast, the predominantly Gs-coupled prostacyclin receptor interacted with Gs, but there was little interaction between the prostacyclin receptor and Go. Inverse agonists did not change the FRET ratio, whereas the addition of agonist resulted in a modest fall. Our work suggests that GPCR dimers and the G protein heterotrimer are present at the cell membrane in the resting state in a pentameric complex. PMID- 16352730 TI - Epicardial retinoid X receptor alpha is required for myocardial growth and coronary artery formation. AB - Vitamin A signals play critical roles during embryonic development. In particular, heart morphogenesis depends on vitamin A signals mediated by the retinoid X receptor alpha (RXRalpha), as the systemic mutation of this receptor results in thinning of the myocardium and embryonic lethality. However, the molecular and cellular mechanisms controlled by RXRalpha signaling in this process are unclear, because a myocardium-restricted RXRalpha mutation does not perturb heart morphogenesis. Here, we analyze a series of tissue-restricted mutations of the RXRalpha gene in the cardiac neural crest, endothelial, and epicardial lineages, and we show that RXRalpha signaling in the epicardium is required for proper cardiac morphogenesis. Moreover, we detect an additional phenotype of defective coronary arteriogenesis associated with RXRalpha deficiency and identify a retinoid-dependent Wnt signaling pathway that cooperates in epicardial epithelial-to-mesenchymal transformation. PMID- 16352731 TI - Epstein-Barr virus latent antigen 3C can mediate the degradation of the retinoblastoma protein through an SCF cellular ubiquitin ligase. AB - Epstein-Barr virus (EBV) stimulates the proliferation of latently infected B cells and promotes lymphoid malignancies in humans. To address the role of EBV latency protein Epstein-Barr nuclear antigen 3C (EBNA3C) in regulation of the retinoblastoma protein (Rb), we transfected EBNA3C into 293, BJAB, and SAOS-2 cells. In this context, a dominant effect of EBNA3C is to decrease Rb protein levels. EBNA3C also rescues an Rb-induced flat cell phenotype and targets Rb for proteasome- and ubiquitin-dependent degradation. Further, EBNA3C forms a stable complex with Rb in cells when the proteasome machinery is inhibited and interacts with Rb in vitro, mapping to a conserved domain at the terminus of EBNA3C. Deletion analysis of EBNA3C identified a motif within amino acids 140-149 important for both the binding and regulation of Rb. This motif is of particular interest, because it has also been linked to regulation of the Skp1/Cul1/F-box complex, SCF(Skp2). Indeed, inhibition of Skp2 function with a dominant-negative molecule reduces the ability of EBNA3C to degrade Rb. Skp2 has no detectable effect on Rb levels in the absence of EBNA3C, suggesting that SCF(Skp2) is specifically usurped by EBNA3C for the enhancement of Rb degradation. That EBNA3C has exploited this association suggests that other human malignancies might use a similar strategy to regulate the Rb protein. PMID- 16352732 TI - Mechanism of metal ion activation of the diphtheria toxin repressor DtxR. AB - The diphtheria toxin repressor (DtxR) is a metal ion-activated transcriptional regulator that has been linked to the virulence of Corynebacterium diphtheriae. Structure determination has shown that there are two metal ion binding sites per repressor monomer, and site-directed mutagenesis has demonstrated that binding site 2 (primary) is essential for recognition of the target DNA repressor, leaving the role of binding site 1 (ancillary) unclear. Calorimetric techniques have demonstrated that although binding site 1 (ancillary) has high affinity for metal ion with a binding constant of 2 x 10(-7), binding site 2 (primary) is a low-affinity binding site with a binding constant of 6.3 x 10(-4). These two binding sites act in an independent fashion, and their contribution can be easily dissected by traditional mutational analysis. Our results clearly demonstrate that binding site 1 (ancillary) is the first one to be occupied during metal ion activation, playing a critical role in stabilization of the repressor. In addition, structural data obtained for the mutants Ni-DtxR(H79A,C102D), reported here, and the previously reported DtxR(H79A) have allowed us to propose a mechanism of metal activation for DtxR. PMID- 16352733 TI - US of the shoulder: rotator cuff and non-rotator cuff disorders. AB - Ultrasonography (US) has been shown to be an effective imaging modality in the evaluation of both rotator cuff and non-rotator cuff disorders, usually serving in a complementary role to magnetic resonance imaging of the shoulder. US technique for shoulder examination depends on patient positioning, scanning protocol for every tendon and anatomic part, and dynamic imaging. The primary US signs for rotator cuff supraspinatus tendon tears are tendon nonvisualization for complete tears, focal tendon defect for full-thickness tears, a hypoechoic defect of the articular side of the tendon for an articular-side partial-thickness tear, and flattening of the bursal surface of the tendon for a bursal-side partial thickness tear. Secondary US signs such as cortical irregularity of the greater tuberosity and joint and subacromial-subdeltoid bursal fluid are helpful when correlated with the primary signs. Tendon degeneration, tendinosis, and intrasubstance tear are demonstrated as internal heterogeneity. Long-head biceps tendon abnormalities include instability, acute or chronic tear, and tendinosis. The acromioclavicular joint is assessed for dislocation, fluid collection, cysts, and bone erosions. Other non-rotator cuff disorders include synovial disorders such as adhesive capsulitis and synovial osteochondromatosis; degenerative disorders such as osteoarthritis, amyloid arthropathy, hemarthrosis, and chondrocalcinosis; infectious disorders such as septic arthritis and bursitis; and space-occupying lesions. PMID- 16352734 TI - Beta-lactam resistance and beta-lactamase expression in clinical Stenotrophomonas maltophilia isolates having defined phylogenetic relationships. AB - AIMS: To test the hypothesis that Stenotrophomonas maltophilia isolates from certain phylogenetic groups have predictable beta-lactamase expression and beta lactam resistance profiles. METHODS: Isolates were grouped using sequences of the 16S rRNA gene and smeT-smeD intergenic region. beta-Lactamase activities in cell extracts were quantified spectrophotometrically and beta-lactam MICs were determined using agar dilution methodology and Etest as appropriate. RESULTS: A collection of 50 clinical S. maltophilia isolates from Europe and North, South and Central America were phylogenetically grouped. Group 'A' (22 out of 50) includes remarkably genetically homogeneous isolates; group 'B' (17 out of 50) includes isolates that are genetically heterogeneous and quite distinct from those of group A. Members of these two groups are, however, indistinguishable in terms of their beta-lactam resistance and beta-lactamase expression phenotypes. In contrast, isolates from group 'C', which are less common (8 out of 50), are considerably more susceptible to beta-lactams owing to reduced inducibility of beta-lactamase expression following beta-lactam challenge. CONCLUSIONS: The majority of S. maltophilia clinical isolates behave similarly in terms of beta lactamase expression and beta-lactam resistance properties, despite considerable phylogenetic variability. PMID- 16352735 TI - Anti-inflammatory effects of moxifloxacin on IL-8, IL-1beta and TNF-alpha secretion and NFkappaB and MAP-kinase activation in human monocytes stimulated with Aspergillus fumigatus. AB - OBJECTIVES: We have previously shown that moxifloxacin conferred protective anti inflammatory effects against Candida pneumonia in immunosuppressed mice. Further in vitro studies showed anti-inflammatory effects of moxifloxacin in LPS and cytokine-stimulated monocytic and epithelial cells. In the present study, concentrating on a more challenging pathogen of immunosuppressed hosts, we studied the effect of moxifloxacin on cytokine secretion and signal transduction mechanisms in monocytic cells stimulated with Aspergillus fumigatus. METHODS: Human peripheral blood monocytes (PBMCs) and a human monocytic cell line (THP-1) were incubated with 1.5x10(6)/mL conidia of a clinical isolate of A. fumigatus. Cytokine secretion and activation of NFkappaB and the MAP-kinases ERK1/2 and p38 were measured with and without the addition of moxifloxacin (5-20 mg/L). RESULTS: Stimulation of PBMCs and THP-1 cells with A. fumigatus increased IL-8, IL-1beta and TNF-alpha secretion (4.1-, 8.3- and 7-fold, and 5.4-, 3.7- and 17.8-fold, respectively). Addition of moxifloxacin (5-20 mg/L) inhibited cytokine secretion up to 45.7+/-5%, 72+/-13% and 73+/-10% in PBMCs and up to 35.6+/-0.5%, 30+/-2.4% and 19+/-4% in THP-1 cells (P<0.05). Signal transduction studies showed that incubation of THP-1 cells with A. fumigatus increased ERK1/2 and p38 phosphorylation and p65-NFkappaB protein expression by 1.6-, 1.3- and 1.8-fold, respectively. Addition of moxifloxacin inhibited ERK1/2, p38 and p65-NFkappaB by up to 69+/-14%, 58+/-3% and 75+/-15%, respectively. CONCLUSIONS: Our results indicate that moxifloxacin acts as an anti-inflammatory agent in monocytic cells stimulated with A. fumigatus conidia. Whether these effects may be protective as in the Candida pneumonia model is unknown and merits in vivo studies in models of pulmonary aspergillosis. PMID- 16352736 TI - Genetic relatedness of a rarely isolated Salmonella: Salmonella enterica serotype Niakhar from NARMS animal isolates. AB - BACKGROUND: In the United States, Salmonella enterica serotype Niakhar is infrequently isolated. Between 1997 and 2000, the animal arm of the National Antimicrobial Resistance Monitoring System-Enteric Bacteria (NARMS) assayed a total of 22,383 Salmonella isolates from various animal sources (swine, cattle, chickens, turkeys, cats, horses, exotics and dogs) for antimicrobial susceptibility. Isolates originated from diagnostic and non-diagnostic submissions. OBJECTIVES: To study the phenotypic and genotypic characteristics of Salmonella Niakhar. METHODS AND RESULTS: Only five (0.02%) of the 22,383 isolates were identified as Salmonella Niakhar. Antimicrobial resistance testing indicated that three isolates were pan-susceptible, one isolate was resistant to ampicillin and one isolate was resistant to ampicillin, chloramphenicol, ciprofloxacin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline and trimethoprim/sulfamethoxazole. RAPD-PCR analysis, PFGE and ribotyping indicated that two pan-susceptible isolates were genetically similar, whereas the three remaining isolates were genetically different. The one Salmonella Niakhar isolate that was multiresistant harboured a class I integron, intI1 and two large plasmids. CONCLUSIONS: This study represents the first report of a ciprofloxacin resistant Salmonella isolate from the animal arm of NARMS. PMID- 16352737 TI - A functional promoter polymorphism in monocyte chemoattractant protein-1 is associated with increased susceptibility to pulmonary tuberculosis. AB - We examined the distribution of single nucleotide polymorphisms (SNPs) in nitric oxide synthase 2A, monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T cell expressed and secreted, and macrophage inflammatory protein-1alpha genes in tuberculosis patients and healthy controls from Mexico. The odds of developing tuberculosis were 2.3- and 5.4-fold higher in carriers of MCP-1 genotypes AG and GG than in homozygous AA. Cases of homozygous GG had the highest plasma levels of MCP-1 and the lowest plasma levels of IL-12p40, and these values were negatively correlated. Furthermore, stimulation of monocytes from healthy carriers of the genotype GG with Mycobacterium tuberculosis antigens yielded higher MCP-1 and lower IL-12p40 concentrations than parallel experiments with monocytes from homozygous AA. Addition of anti-MCP-1 increased IL-12p40 levels in cultures of M. tuberculosis-stimulated monocytes from homozygous GG, and addition of exogenous MCP-1 reduced IL-12p40 production by M. tuberculosis stimulated monocytes from homozygous AA. Furthermore, we could replicate our results in Korean subjects, in whom the odds of developing tuberculosis were 2.8- and 6.9-fold higher in carriers of MCP-1 genotypes AG and GG than in homozygous AA. Our findings suggest that persons bearing the MCP-1 genotype GG produce high concentrations of MCP-1, which inhibits production of IL-12p40 in response to M. tuberculosis and increases the likelihood that M. tuberculosis infection will progress to active pulmonary tuberculosis. PMID- 16352738 TI - Secretory leucoprotease inhibitor binds to NF-kappaB binding sites in monocytes and inhibits p65 binding. AB - Secretory leucoprotease inhibitor (SLPI) is a nonglycosylated protein produced by epithelial cells. In addition to its antiprotease activity, SLPI has been shown to exhibit antiinflammatory properties, including down-regulation of tumor necrosis factor alpha expression by lipopolysaccharide (LPS) in macrophages and inhibition of nuclear factor (NF)-kappaB activation in a rat model of acute lung injury. We have previously shown that SLPI can inhibit LPS-induced NF-kappaB activation in monocytic cells by inhibiting degradation of IkappaBalpha without affecting the LPS-induced phosphorylation and ubiquitination of IkappaBalpha. Here, we present evidence to show that upon incubation with peripheral blood monocytes (PBMs) and the U937 monocytic cell line, SLPI enters the cells, becoming rapidly localized to the cytoplasm and nucleus, and affects NF-kappaB activation by binding directly to NF-kappaB binding sites in a site-specific manner. SLPI can also prevent p65 interaction with the NF-kappaB consensus region at concentrations commensurate with the physiological nuclear levels of SLPI and p65. We also demonstrate the presence of SLPI in nuclear fractions of PBMs and alveolar macrophages from individuals with cystic fibrosis and community-acquired pneumonia. Therefore, SLPI inhibition of NF-kappaB activation is mediated, in part, by competitive binding to the NF-kappaB consensus-binding site. PMID- 16352739 TI - Mastocytosis in mice expressing human Kit receptor with the activating Asp816Val mutation. AB - Mastocytosis is a rare neoplastic disease characterized by a pathologic accumulation of tissue mast cells (MCs). Mastocytosis is often associated with a somatic point mutation in the Kit protooncogene leading to an Asp/Val substitution at position 816 in the kinase domain of this receptor. The contribution of this mutation to mastocytosis development remains unclear. In addition, the clinical heterogeneity presented by mastocytosis patients carrying the same mutation is unexplained. We report that a disease with striking similarities to human mastocytosis develops spontaneously in transgenic mice expressing the human Asp816Val mutant Kit protooncogene specifically in MCs. This disease is characterized by clinical signs ranging from a localized and indolent MC hyperplasia to an invasive MC tumor. In addition, bone marrow-derived MCs from transgenic animals can be maintained in culture for >24 mo and acquire growth factor independency for proliferation. These results demonstrate a causal link in vivo between the Asp816Val Kit mutation and MC neoplasia and suggest a basis for the clinical heterogeneity of human mastocytosis. PMID- 16352740 TI - Suppression of tumor formation in lymph nodes by L-selectin-mediated natural killer cell recruitment. AB - Natural killer (NK) cells are known to reject certain tumors in vivo; however, the ability of NK cells to prevent metastasis of tumors into secondary lymphoid organs has not been addressed. Here, we report that in tumor-bearing hosts, NK cells are recruited to regional lymph nodes in wild-type mice, but not in mice deficient for L-selectin or L-selectin ligands. By adoptive transfer and complete Freund's adjuvant stimulation experiments, we demonstrated that L-selectin on NK cells and L-selectin ligands on endothelial cells are essential for NK cell recruitment to lymph nodes. Furthermore, freshly isolated resident lymph node NK cells lysed tumors efficiently, and metastasis of B16 melanoma cells to draining lymph nodes was suppressed in wild-type or Rag-1-deficient mice, but not when NK cells were depleted. Although L-selectin-deficient NK cells efficiently lysed tumor cells in vitro, NK cell-dependent suppression of tumor metastasis was diminished in mice deficient for L-selectin or L-selectin ligands because of insufficient NK cell recruitment to lymph nodes. Moreover, tumor metastasis was substantially inhibited in L-selectin-deficient mice reconstituted with wild-type NK cells. These findings indicate that L-selectin-mediated NK cell recruitment plays a crucial role in the control of tumor metastasis into secondary lymphoid organs. PMID- 16352741 TI - Hydronephrosis associated with antiurothelial and antinuclear autoantibodies in BALB/c-Fcgr2b-/-Pdcd1-/- mice. AB - Because most autoimmune diseases are polygenic, analysis of the synergistic involvement of various immune regulators is essential for a complete understanding of the molecular pathology of these diseases. We report the regulation of autoimmune diseases by epistatic effects of two immunoinhibitory receptors, low affinity type IIb Fc receptor for IgG (FcgammaRIIB) and programmed cell death 1 (PD-1). Approximately one third of the BALB/c-Fcgr2b(-/-)Pdcd1(-/-) mice developed autoimmune hydronephrosis, which is not observed in either BALB/c Fcgr2b(-/-) or BALB/c-Pdcd1(-/-) mice. Hydronephrotic mice produced autoantibodies (autoAbs) against urothelial antigens, including uroplakin IIIa, and these antibodies were deposited on the urothelial cells of the urinary bladder. In addition, approximately 15% of the BALB/c-Fcgr2b(-/-)Pdcd1(-/-) mice produced antinuclear autoAbs. In contrast, the frequency of the autoimmune cardiomyopathy and the production of anti-parietal cell autoAb, which were observed in BALB/c-Pdcd1(-/-) mice, were not affected by the additional FcgammaRIIB deficiency. These observations suggest cross talk between two immunoinhibitory receptors, FcgammaRIIB and PD-1, on the regulation of autoimmune diseases. PMID- 16352742 TI - Molecular analysis of impaired urinary diluting capacity in glucocorticoid deficiency. AB - Urinary diluting ability and protein abundance of renal aquaporins (AQPs) and ion transporters in glucocorticoid-deficient (GD) rats were examined at baseline and in response to oral water loading. Rats underwent bilateral adrenalectomy followed by aldosterone (GD) or aldosterone + dexamethasone (CTL) replacement. Before oral water loading, urinary output was significantly decreased and urinary osmolality (U(osm)) was increased in GD compared with CTL rats. Protein abundance of inner medullary AQP2 (148 +/- 18%), phosphorylated AQP2 (pAQP2, 156 +/- 13%), and AQP3 (145 +/- 8%) was significantly upregulated in GD compared with CTL rats (all P < 0.05). GD rats also demonstrated a marked reduction in urinary Na(+) excretion compared with pair-fed CTL rats. Na(+)-K(+)-2Cl(-) cotransporter, Na(+)/H(+) exchanger type 3, and cortical beta- and gamma-subunits of the epithelial Na(+) channel were significantly upregulated in GD rats. At 1 h after an acute water load (40 ml/kg by oral gavage), GD rats demonstrated a decrease in percent water excretion (5 +/- 1 vs. 33 +/- 9%, P < 0.01) and urinary output (33 +/- 12 vs. 250 +/- 65 microl x kg(-1) x min(-1), P < 0.05) and an increase in U(osm) (1,894 +/- 292 vs. 316 +/- 92 mosmol/kgH(2)O, P < 0.001) compared with CTL rats. Plasma AVP was increased (1.6 +/- 0.2 vs. 0.9 +/- 0.2 pg/ml, P < 0.05), as was protein expression of inner medullary AQP2 (149 +/- 5%) and pAQP2 (177 +/- 9%, P < 0.01), in GD compared with CTL rats; apical expression of AQP2 was maintained in GD rats. The vasopressin V(2) receptor antagonist OPC-31260 increased percent water excretion and urinary output and reduced U(osm) compared with vehicle-treated GD rats. OPC-31260 also reversed the increased abundance and apical trafficking of inner medullary AQP2 and pAQP2 protein in GD rats. In conclusion, enhanced protein abundance of Na(+) transporters and Na(+) channels with Na(+) retention occurred with GD. OPC-31260 reversed upregulation and apical trafficking of AQP2 and pAQP2 in association with improved urinary diluting capacity and increased water excretion after oral water loading. PMID- 16352743 TI - Kidney oxygen consumption, carbonic anhydrase, and proton secretion. AB - Oxygen consumed by the kidney (Q(O(2))) is primarily obligated to sodium reabsorption (T(Na)). The relationship of Q(O(2)) to T(Na) (Q(O(2))/T(Na)) may be altered by hormones and autacoids. To examine whether Q(O(2))/T(Na) depends on the mechanism of sodium reabsorption, we first evaluated the effects on Q(O(2)) and Q(O(2))/T(Na) of benzolamide (BNZ), a proximal diuretic that works by inhibiting membrane carbonic anhydrase. During BNZ infusion in anesthetized rats, Q(O(2)) increased by 50% despite a 25% decline in T(Na). However, BNZ failed to increase Q(O(2))/T(Na) when given along with the adenosine A1 receptor blocker, DPCPX, which inhibits basolateral Na-bicarbonate cotransport (NBC1), or EIPA, which inhibits sodium-hydrogen exchange (NHE). Incubating freshly harvested rat proximal tubules with BNZ also caused Q(O(2))to increase by 62%, an effect that was prevented by blocking the apical NHE3 with S3226. Blocking NBC1 or NHE3 in the proximal tubule will have opposite effects on cell pH, but both maneuvers should reduce active chloride transport. In conclusion, inhibiting membrane carbonic anhydrase in the proximal tubule increases Q(O(2)) and reduces the energy efficiency of sodium reabsorption by the kidney. This is not purely due to shifting the burden of reabsorption to a more expensive site downstream from the proximal tubule. Instead, increased cost may be incurred within the proximal tubule as the result of increased active chloride transport. PMID- 16352744 TI - Nitric oxide induces apoptosis in renal tubular epithelial cells through activation of caspase-8. AB - The susceptibility or resistance of tubular epithelial cells (TEC) to apoptosis is pivotal to the long-term maintenance of kidney function following episodes of inflammation, such as graft rejection. TEC apoptosis can occur with ischemia as well as with proinflammatory cytokines and nitric oxide (NO), produced by infiltrating mononuclear cells. TEC can also produce abundant amounts of NO during inflammation but the role and regulation of NO-induced injury of TEC are not well understood. Apoptosis in TEC in vitro was determined by FACS analysis with annexin-V and propidium iodide staining. NO in culture supernatants was measured by Greiss reagent, and protein expression of inducible NO synthetase (NOS2/iNOS) and caspase-8 was examined by Western blot analysis. Here, we showed that murine TEC produced abundant amounts of NO in response to proinflammatory cytokines (IFN-gamma/TNF-alpha) through upregulation of NOS2, and inhibition of endogenous NO production by l-NMMA reduced TEC apoptosis in cytokine-stimulated cultures. Addition of exogenous NO (sodium nitroprusside) induced TEC apoptosis as well as caspase-8 activation in a dose-dependent manner. The key role of caspase-8 in NO-induced TEC apoptosis was demonstrated by that NO-induced TEC apoptosis can be blocked by caspase-8 inhibition using z-IETD-fmk, caspase-8 silencing with shRNA or by overexpressing the endogenous caspase-8 inhibitor c FLIP (cellular Flice-inhibitory protein). In conclusion, endogenous NO from NOS2 activity as well as exogenous NO can contribute to renal injury through apoptosis of TEC. Activation of caspase-8 plays a central role in NO-induced apoptosis and caspase-8 inhibition may be an important therapeutic target during renal inflammation. PMID- 16352745 TI - Cardiac glycoside downregulates NHE3 activity and expression in LLC-PK1 cells. AB - Ouabain, a cardiotonic steroid and a specific inhibitor of the Na(+)-K(+)-ATPase, has been shown to significantly inhibit transcellular Na(+) transport without altering the intracellular Na(+) concentration ([Na(+)](i)) in the epithelial cells derived from the renal proximal tubules. We therefore studied whether ouabain affects the activity and expression of Na(+)/H(+) exchanger isoform 3 (NHE3) representing the major route of apical Na(+) reabsorption in LLC-PK(1) cells. Chronic basolateral, but not apical, exposure to low-concentration ouabain (50 and 100 nM) did not change [Na(+)](i) but significantly reduced NHE3 activity, NHE3 protein, and mRNA expression. Inhibition of c-Src or phosphoinositide 3-kinase (PI3K) with PP2 or wortmannin, respectively, abolished ouabain-induced downregulation of NHE3 activity and mRNA expression. In caveolin 1 knockdown LLC-PK(1) cells, ouabain failed to downregulate NHE3 mRNA expression and NHE3 promoter activity. Ouabain response elements were mapped to a region between -450 and -1,194 nt, where decreased binding of thyroid hormone receptor (TR) and Sp1 to their cognate cis-elements was documented in vitro and in vivo by protein/DNA array analysis, EMSA, supershift, and chromatin immunoprecipitation. These data suggest that, in LLC-PK(1) cells, ouabain-induced signaling through the Na(+)-K(+)-ATPase-Src pathway results in decreased Sp1 and TR DNA binding activity and consequently in decreased expression and activity of NHE3. These novel findings may represent the underlying mechanism of cardiotonic steroid mediated renal compensatory response to volume expansion and/or hypertension. PMID- 16352746 TI - The immunophilin FKBP52 inhibits the activity of the epithelial Ca2+ channel TRPV5. AB - In the kidney, the epithelial Ca(2+) channel TRPV5 constitutes the apical entry pathway in the process of active Ca(2+) reabsorption. The regulation of Ca(2+) influx through TRPV5 is of crucial importance, because it determines the final amount of Ca(2+) excreted in the urine. The present study identifies FKBP52 as an auxiliary protein of TRPV5, inhibiting channel activity. FKBP52 shows specific interaction with TRPV5, and both proteins colocalize in the distal part of the nephron. On the functional level, FKBP52 decreases Ca(2+) influx through TRPV5 as demonstrated in radioactive (45)Ca(2+) uptake measurements and electrophysiological studies in TRPV5-overexpressing human embryonic kidney 293 cells. On the other hand, gene silencing of FKBP52 or administration of the FKBP52 blocker FK-506 enhances Ca(2+) influx through TRPV5. The inhibitory action of FKBP52 on TRPV5 activity is blunted by mutation of its peptidyl-propyl cis trans isomerase domain, showing that the FKBP52 catalytic property is critical for channel activity. In conclusion, these results suggest that FKBP52 plays an important role in the regulation of TRPV5 and thus in the process of Ca(2+) reabsorption. PMID- 16352747 TI - Vasopressin induces expression of the Cl-/HCO3- exchanger SLC26A7 in kidney medullary collecting ducts of Brattleboro rats. AB - SLC26A7 is a newly identified basolateral Cl(-)/HCO(3)(-) exchanger specific to alpha-intercalated cells of the outer medullary collecting duct (OMCD). The purpose of the present experiments was to examine the expression of SLC26A7 in kidneys of vasopressin-deficient Brattleboro rats before and after treatment with desamino-Cys(1),d-Arg(8)-vasopressin (dDAVP). Brattleboro rats were treated with dDAVP, a vasopressin analog, for 8 days, and their kidneys were examined for the expression of SLC26A7. The expression of SLC26A7 protein, as examined by immunofluorescence, was undetectable in kidneys of Brattleboro rats. However, treatment with dDAVP induced expression of SLC26A7 protein, restoring it to levels observed in normal rats. These results were verified by Western blot analysis. The mRNA expression of SLC26A7 remained unchanged in response to dDAVP. Immunofluorescent labeling demonstrated abundant levels of anion exchanger type 1 in the OMCD of Brattleboro rats and a mild reduction in response to dDAVP. The abundance of H(+)-ATPase was not affected by dDAVP. The increased SLC26A7 expression directly correlated with enhanced aquaporin-2 expression, which is proportional to increased interstitial osmolarity in the medulla. In conclusion, vasopressin increases the expression of SLC26A7 protein through posttranscriptional mechanisms in the OMCD. The induction of SLC26A7 by vasopressin in OMCD cells of Brattleboro rats is likely an attempt by cells to regulate their cell volume and maintain HCO(3)(-) absorption in a state associated with increased interstitial medullary tonicity. PMID- 16352748 TI - Clinical year in review IV: asthma, chronic obstructive pulmonary disease, exercise and rehabilitation, and critical care medicine. PMID- 16352750 TI - The evolution and state-of-the-art principles of multislice computed tomography. AB - Computed tomography has revolutionized diagnostic radiology with the introduction of spiral CT in the early 1990s, allowing for the first time the acquisition of volume data without the danger of misregistration or double registration of anatomical details. The next revolution occurred in 1998 when all major CT manufacturers introduced multislice CT (MSCT) systems, which typically offered simultaneous acquisition of four slices, providing considerable improvement towards the goal of isotropic three-dimensional imaging. The most recent generation of MSCT systems acquire 64 slices per rotation, enabling a whole body CTA with 1,500 mm scan range and an isotropic resolution of down to 0.4 mm in only 22- 25 s. The tube and the detector measurement system belong to the most important system components of a CT system, having a large influence on system performance. For example, new rotating envelope tube design principles allow for faster rotation times and for double z-sampling techniques in order to increase resolution. The applied dose is ultimately the limiting factor for the improvement of image quality and increase in isotropic resolution. In order to make best diagnostic use of the applied dose, sophisticated dynamic dose adaptation techniques to patient size and geometry have been developed. PMID- 16352751 TI - Micro-computed tomography of the lungs and pulmonary-vascular system. AB - Three-dimensional imaging of the intact lung and its vasculature is essential if the hierarchical and volumetric aspects of its structures and functions are to be quantitated. Although this is possible with clinical multislice helical CT scanners, the spatial resolution does not scale down adequately for small rodents for which cubic voxel dimensions of 50-100 microm are required. Micro-computed tomography (micro-CT) provides the necessary spatial resolution of 3D images of the intact thoracic contents. Micro-CT can provide higher resolution so that basic micro-architectural structures, such as alveoli, can be individually visualized and quantitated. Dynamic events, such as the respiratory and cardiac cycles, can be imaged at multiple time points throughout a representative cycle by coordinating the scan sequence (i.e., gating) to the cycle phase of a sequence of cycles. Fusion of the micro-CT image data with other image data, such as micro SPECT or histology, can enhance the information content beyond the mainly structural information provided by micro-CT. Conventional attenuation-based X-ray imaging can involve significant X-ray exposures at high spatial resolutions, and this could affect the phenotype (e.g., via interstitial fibrosis) and genotype (e.g., via mutation), so its use in longitudinal studies using micro-CT may be limited in some cases. However, because of recent developments in which the phase shift or refraction of X-rays rather than attenuation is used, the X-ray exposure may be significantly reduced. PMID- 16352752 TI - Morphology of the small-animal lung using magnetic resonance microscopy. AB - Small-animal imaging with magnetic resonance microscopy (MRM) has become an important tool in biomedical research. When MRM is used to image perfusion-fixed and "stained" whole mouse specimens, cardiopulmonary morphology can be visualized, nondestructively, in exquisite detail in all three dimensions. This capability can be a valuable tool for morphologic phenotyping of different mouse strains commonly used in genomics research. When these imaging techniques are combined with specialized methods for biological motion control and animal support, the lungs of the live, small animal can be imaged. Although in vivo imaging may not achieve the high resolution possible with a fixed specimen, dynamic functional studies and survival studies that follow the progression of pulmonary change related to disease or environmental exposure are possible. By combining conventional proton imaging with gas imaging, using hyperpolarized 3He, it is possible to image the tissue and gas compartments of the lung. This capability is illustrated in studies on an emphysema model in rats and on radiation damage of the lung. With further improvements in imaging and animal handling technology, we will be able to image faster and at higher resolutions, making MRM an even more valuable research tool. PMID- 16352753 TI - Segmentation and quantitative analysis of intrathoracic airway trees from computed tomography images. AB - The segmentation of the human airway tree from volumetric multidetector-row computed tomography images is an important prerequisite for many clinical applications and physiologic studies. We present a new airway segmentation method based on fuzzy connectivity. Small adaptive regions of interest are used that follow the airway branches as they are segmented. This method works on various types of scans (low dose and regular dose, normal subjects and diseased subjects) without the need for the user to manually adjust any parameters. Comparison with a commonly used region-growing segmentation algorithm shows that this method retrieves a significantly higher count of airway branches. In an additional processing step, this method provides accurate cross-sectional airway measurements that are conducted in the original gray-level volume. Validation on a phantom shows that subvoxel accuracy is achieved for all airway sizes and airway orientations. The utility of the reported method is demonstrated in a comparative analysis of normal and cystic fibrosis airway trees. PMID- 16352754 TI - Virtual bronchoscopy. AB - Virtual bronchoscopy is rapidly providing a software solution for visualization and measurement of the human airway tree with data derived from multirow detector X-ray CT scans as the most common data source. Increasingly, other imaging data sources to create three-dimensional image renderings of the bronchial tree are being used including magnetic resonance imaging, ultrasound, and the digital color image taken by the bronchoscope itself. Software solutions now exist for providing simple renderings of the bronchial tree through which a fly-through of the airway lumen by the center line of the airway can be added (the fly-through mimics the view that a real flexible bronchoscope affords the operator). The images so produced are visually accurate (i.e., they appear very realistic) and with good software now available are also analytically correct (i.e., measurements taken from these images are accurate). More advanced virtual bronchoscopic applications, including image-based pathfinding to mediastinal and peripheral lung structures, are also in development. Synergistic datasets composed of image data from multiple image sources are also being developed. PMID- 16352755 TI - Computed tomography studies of lung ventilation and perfusion. AB - With the emergence of multidetector-row computed tomography (CT) it is now possible to image both structure and function via use of a single imaging modality. Breath-hold spiral CT provides detail of the airway and vascular trees along with texture reflective of the state of the lung parenchyma. Use of stable xenon gas wash-in and/or wash-out methods using an axial mode of the CT scanner whereby images are acquired through gating to the respiratory cycle provide detailed images of regional ventilation with isotropic voxel dimensions now on the order of 0.4 mm. Axial scanning during a breath hold and gating to the electrocardiogram during the passage of a sharp bolus injection of iodinated contrast agent provide detailed images of regional pulmonary perfusion. These dynamic CT methods for the study of regional lung function are discussed in the context of other methods that have been used to study heterogeneity of lung function. PMID- 16352757 TI - Computed tomography studies of lung mechanics. AB - The study of lung mechanics has progressed from global descriptions of lung pressure and volume relationships to the high-resolution, three-dimensional, quantitative measurement of dynamic regional mechanical properties and displacements. X-ray computed tomography (CT) imaging is ideally suited to the study of regional lung mechanics in intact subjects because of its high spatial and temporal resolution, correlation of functional data with anatomic detail, increasing volumetric data acquisition, and the unique relationship between CT density and lung air content. This review presents an overview of CT measurement principles and limitations for the study of regional mechanics, reviews some of the early work that set the stage for modern imaging approaches and impacted the understanding and management of patients with acute lung injury, and presents evolving novel approaches for the analysis and application of dynamic volumetric lung image data. PMID- 16352758 TI - Positron emission tomography imaging of regional pulmonary perfusion and ventilation. AB - Positron emission tomography (PET) imaging is a noninvasive, quantitative method to assess pulmonary perfusion and ventilation in vivo. The core of this article focuses on the use of [13N]nitrogen (13N2) and PET to assess regional gas exchange. Regional perfusion and shunt can be measured with the 13N2-saline bolus infusion technique. A bolus of 13N2, dissolved in saline solution, is injected intravenously at the start of a brief apnea, while the tracer kinetics in the lung is measured by a sequence of PET frames. Because of its low solubility in blood, virtually all 13N2 delivered to aerated lung regions diffuses into the alveolar airspace, where it accumulates in proportion to regional perfusion during the apnea. In contrast, lung regions that are perfused but are not aerated and do not exchange gas (i.e., "shunting" units) do not retain 13N2 during apnea and the tracer concentration drops after the initial peak. Accurate estimates of regional perfusion and regional shunt can be derived by applying a mathematical model to the pulmonary kinetics of a 13N2-saline bolus. When breathing is resumed, specific alveolar ventilation can be calculated from the tracer washout rate, because 13N2 is eliminated almost exclusively by ventilation. Because of the rapid elimination of the tracer, 13N2 infusion scans can be followed by 13N2 inhalation scans that allow determination of regional gas fraction. This article describes insights into the pathophysiology of acute lung injury, pulmonary embolism, and asthma that have been gained by PET imaging of regional gas exchange. PMID- 16352759 TI - Hyperpolarized 3-helium magnetic resonance imaging to probe lung function. AB - Understanding pulmonary pathophysiology has increasing implications for imaging modalities. Although it was sufficient to perform high-resolution computed tomography in the past, the impetus now is on providing quantitative and functional lung data. Magnetic resonance imaging, which was traditionally difficult to perform in the lungs, has developed into a promising technology. One of the main areas of interest is the use of hyperpolarized noble gases, such as 3 He and 129-Xe, which enable high-definition lung imaging that includes information on lung microstructure. It is possible to obtain three-dimensional information on essential pulmonary processes, such as ventilation, oxygen uptake, and spirometry, which offers new insight into lung pathophysiology. This article focuses on the novel aspects of hyperpolarized 3-He magnetic resonance imaging. PMID- 16352760 TI - Instrumentation for molecular imaging in preclinical research: Micro-PET and Micro-SPECT. AB - Noninvasive imaging of molecular events and interactions in living small animal models has gained increasing importance in preclinical research. Two of the imaging modalities available for this research with potential for translation to the clinic are dedicated small animal positron emission tomography and single photon emission computed tomography. This brief review introduces the fundamental principles behind these imaging technologies and instrumentation, and discusses the limitations in terms of their spatial resolution and sensitivity. In addition, it provides a perspective regarding the research and commercial development of these systems and presents examples of biological applications. Finally, it discusses the major challenges facing these technologies, advantages and limitations with respect to other technologies, and some future prospects. PMID- 16352761 TI - Bioluminescence imaging. AB - Bioluminescence refers to the process of visible light emission in living organisms. Bioluminescence imaging is a powerful methodology that has been developed over the last decade as a tool for molecular imaging of small laboratory animals, enabling the study of ongoing biological processes in vivo. This form of optical imaging is low cost and noninvasive and facilitates real time analysis of disease processes at the molecular level in living organisms. In this article, we provide a brief introduction to bioluminescence imaging technology and discuss its applications in mouse models of lung inflammation/injury, bacterial pneumonia, and tumor growth and metastasis. PMID- 16352762 TI - Advances in positron emission tomographic imaging of lung cancer. AB - Positron emission tomography (PET) with [18F]fluorodeoxyglucose (FDG) has been established as a useful tool in the management of patients with non-small cell lung cancer and promises to be as valuable in the clinical management of other cancers. PET imaging with FDG allows the assessment of tumor glucose metabolism in vivo; however, a number of other PET tracers are being used in oncologic research to assess changes in other cellular processes associated with malignant transformation of the cell. [11C]-Labeled methionine and choline are being used to assess changes in cell membrane synthesis; however, small studies have not shown the added information from these tracers to be clinically useful. DNA synthesis can be assessed by measuring the uptake of the thymidine analog 3' deoxy-3'-[18F]fluorothymidine, which may be more specific for evaluating malignancy without the problem of false-positive results from inflammatory lesions, as seen with FDG. Tumor hypoxia imaging with copper-labeled diacetyl bis(N(4)-methylthiosemicarbazone) or [18F]fluoromisonidazole may provide a better method of predicting which tumors will respond best to conventional therapy. The role of PET will continue to evolve with further clinical studies using these and other new tracers. PMID- 16352763 TI - Inflammation imaging. AB - Acute and chronic lung diseases are almost invariably associated with some degree of inflammation. Cells that evolved as an effective mechanism to counter infection and heal lung tissue may, in some circumstances, themselves be partially responsible for the pathogenesis of chronic lung disease that leads to irreversible lung damage and loss of lung function. Although standard measurements of lung function can document the progression of disease, the contributions of the numerous interacting elements to the process are difficult to measure in life. The use of molecular imaging techniques allows the different components of the inflammatory response to be monitored in situ in humans. In particular, positron emission tomography of selected markers targeted to specific cells and biochemical pathways can provide accurate measurements of disease activity, enabling a better understanding of inflammatory processes at all stages of disease. The practicability of sequential measurements allows one to monitor the natural history of different lung diseases. More importantly, imaging provides a unique tool for quantification of the modulation of discrete and specific aspects of inflammatory lung disease by targeted interventions. This should facilitate the development of new treatment strategies with better specificity for key elements of each disease. PMID- 16352765 TI - Deconstruction of the curriculum: a modest proposal. PMID- 16352764 TI - Molecular imaging of pulmonary gene expression with positron emission tomography. AB - Gene expression imaging is one form of molecular imaging used to visualize, characterize, and quantify, spatially and temporally, normal as well as pathologic processes at cellular and subcellular levels within intact living organisms. Most studies to date have employed positron emission tomography as the imaging platform to detect, monitor, and quantify gene expression in the lungs. These studies have shown that imaging can be used to determine the onset and duration of transgene expression, the effectiveness of different gene delivery systems, and the linearity of vector dose-response relationships. This rapidly developing field can be expected to provide useful new tools with which to study gene expression in transgenic animals and in humans during gene therapy. PMID- 16352766 TI - Students' comfort level in treating vulnerable populations and future willingness to treat: results prior to extramural participation. AB - This study analyzed senior dental students' perceptions prior to extramural rotations for comfort and future willingness to treat patients with special needs and other vulnerable groups. The sample included 690 University of Iowa senior dental students who graduated from 1992 through 2004. These students completed a questionnaire concerning twelve vulnerable population groups. Logistic regression models were performed, using student comfort and future willingness to treat each group as the dependent variable. There was a wide percentage of range of comfort with these groups, yet there was no individual group that more than 60 percent of these students were willing to treat in their future practices. Generally, prior experience with the group had a positive impact on comfort level. When gender was included in the regression models, male students were more likely to express comfort. In all instances except one, experience had a positive influence on perceived future willingness to treat the associated group. However, younger graduates had a greater willingness to treat. When controlling for other variables within the future willingness to treat models, comfort was statistically significant only for HIV+/AIDS and non-English speaking groups. This study provides insight about comfort with and perceived future willingness to treat special needs and other vulnerable patient groups. PMID- 16352767 TI - An evidence-based model of effective self-assessment for directing professional learning. AB - An innovative model for conducting meaningful self-assessments is presented to help oral health care professionals efficiently determine what to learn with the goal of remaining competent. A review and analysis of the literature drawing from several databases was conducted to develop the model. Through this process, we identified four key categories: prerequisite competencies, process, applications, and tools that are suggested to occur within a supportive environment to carry out valid self-assessments and to positively influence learning choices and practice improvements. PMID- 16352768 TI - Clinical competence of graduates of community-based and traditional curricula. AB - As community-based dental education programs proliferate, so do concerns about the quality of education at extramural sites. While there have been some comparisons of the effects of community-based programs on attitudes and perceptions of competency, there is virtually no information about whether such programs can train students to carry out standard dental procedures as well as if they were prepared in the dental school proper. To address this gap in our knowledge about the educational outcomes of community-based programs, the following performance measures were retrospectively compared for 457 students in traditional senior year curricula and fifty seniors in a community-based program: scores and pass/fail rates on the Northeastern Regional Boards (NERB), clinical production, passage of competencies, and on-time graduation rates. Students in the CODE (Community-Oriented Dental Education) program at the University of Medicine and Dentistry of New Jersey (UMDNJ)-New Jersey Dental School (NJDS) demonstrated significantly higher scores on the restorative section of the NERB, but this did not translate into a better overall pass rate. CODE students passed the same competencies as graduates of the traditional program, yet demonstrated appreciably higher clinical productivity and on-time graduation rates. The additional clinical experiences for the CODE students may be a factor in their higher board scores. Based on these data, it is concluded that community-based dental educational programs can be at least as effective as intramural (school based) educational experiences in providing students with a sound clinical education. PMID- 16352769 TI - Knowledge, beliefs, and attitudes of dental and dental hygiene students toward obesity. AB - Obesity is recognized as a growing public health problem. The authors surveyed dental hygiene and dental students from one institution regarding education, knowledge, perceived professional duties, and attitudes toward the overweight and obese population. Half of the respondents reported no obesity education prior to professional dental education, and 80 percent received five hours or less while in professional training. While most students held a generally positive attitude regarding obese and overweight patients, a number of students demonstrated evidence of negative stereotyping. Obesity education and training must be integrated into dental education to permit greater understanding of coexisting medical problems, explore the basis for a negative attitude and work toward its elimination, and raise public health awareness within dentistry. PMID- 16352770 TI - The need for tobacco education: studies of collegiate dental hygiene patients and faculty. AB - The need for inclusion of comprehensive tobacco control education/training for health care providers continues to be stressed in publications addressing cessation services. The dental appointment presents an excellent opportunity to provide tobacco interventions to basically healthy people on regular intervals. The purpose of this study was twofold: 1) to assess the need (stage of change and concomitant need for tobacco cessation intervention) of dental hygiene patients at a Midwest dental hygiene clinic, and 2) to assess and compare the level of tobacco intervention education currently being offered by dental hygiene educators in a Midwestern state. Patients (n=426) of a collegiate dental health clinic completed a survey that assessed the level and type of tobacco cessation intervention patients might require. A statewide sample of dental hygiene faculty (n=97) were surveyed to determine the attitudes, perceived barriers, and current practices in tobacco education offered in their programs. Of patients who currently smoked (34.5 percent), 24.7 percent indicated being in the Action stage of change; 14.2 percent were in Preparation; 22.2 percent were in Contemplation; and 29 percent were in Precontemplation. Although faculty indicated tobacco education was very important (5.03 on 1-6 scale), they felt only moderately confident delivering tobacco education (3.18 on a 1-5 scale). Only 16 percent to 35 percent of faculty reported that their curriculum included brief motivational interviewing, pharmacotherapies, or setting-up a private practice tobacco control program. The results strongly suggest the need for a comprehensive, competency based tobacco curriculum to enhance and expand existing dental hygiene programs. PMID- 16352771 TI - Early intervention surveillance strategies (EISS) in dental student clinical performance: a mathematical approach. AB - Graduating dental practitioners requires the mastery of a number of skills and a significant body of basic information. Dental education is a complex combination of didactic and physical skill learning processes. It is necessary to develop appropriate tools to measure student clinical performance to allow the provision of interventional strategies at the right time targeted at the right individuals. In this study, an approach to early intervention surveillance strategies was developed that is cost-effective, transparent, and robust based on mathematical predictions of student clinical achievements. Using a cohort of students' clinical activity profile, a polynomial pair was developed that represents the predictive function of low and high achieving students. This polynomial pair can then be applied to students to predict their final achievement based on their current status. The polynomial methodology is adaptable to local variation such as access to clinical facilities. The early intervention surveillance strategy developed in this study provides a simple, cost-effective, predictive risk assessment system that relies on data sets already collected in most dental schools and can be completed without the need for significant human intervention. The mathematical approach allows the focusing of educational support towards students that require the assistance, thus augmenting the better use of resources. PMID- 16352772 TI - Dental school applicants' use of website information during the application process. AB - The University of Pittsburgh's School of Dental Medicine (SDM) is a medium-sized, private, state-related dental school. As other schools have done recently, the SDM made a substantial effort to upgrade its website in 2003. Internal satisfaction and anecdotal data notwithstanding, there is no measure of the website's usefulness in attracting applicants. We designed an anonymous survey of twenty-one questions in an effort to better understand what kinds of information sources our applicants used to make their application decisions; which information they perceived as useful during each phase of the overall application process; how they ranked our website compared to those of other dental schools that they considered; and what they thought could make our website better. Our study supports the notion that applicants rely in part on dental school websites during the application process. Data from this study point to the areas that are of most interest to applicants when they visit a dental school website. This information could help dental school administrators determine the effort they should spend on their school's website as an advertising and recruitment tool. We recommend that dental schools make the most important application-related information and criteria easy to find. PMID- 16352773 TI - Insights from students following an educational rotation through dental geriatrics. AB - Little is known about how dental students respond to dental geriatrics. This article describes a qualitative analysis of reflective journals submitted over two years by ninety-two senior students who participated in a brief clinical rotation in long-term care facilities. We used an inductive interpretive approach to analyze the journals. Eight themes emerged from the analysis: 1) complexity of the institutional environment; 2) heterogeneity of the resident population; 3) multidisciplinary environment; 4) record keeping; 5) interactions with residents; 6) the difficulty of oral health care for frail residents; 7) bridging the gap between theory and practice; and 8) the emotional impact of the clinical experiences. Apparently, the students appreciated the opportunity to witness the complexity of care in a multidisciplinary context and to observe a practical program of oral health care. They described the rotations as unique and emotionally challenging but very worthwhile. Overall, they wrote positively about their experiences with the elderly residents, acknowledged the contribution of the rotation as important to their clinical maturation, and reported that the experience enhanced their appreciation of a dentist's professional responsibilities. PMID- 16352774 TI - Dental clinical teaching: perceptions of students and teachers. AB - The objective of this study was to explore perceptions of dental student clinicians and clinical teachers about dental clinical teaching to provide primary data for dental researchers and educators. Student focus group data provided background for development of a questionnaire that explored three themes related to clinical teaching. Twenty-one teachers and forty-five student respondents completed the twenty-five-item questionnaire in 2003. In the theme of the teacher/student relationship, no statistically significant differences were seen between teacher and student group perceptions. In the theme of educational theory applied in dental clinical teaching, a statistically significant difference was seen between teacher and student groups in the value of preclinical instruction in senior clinical years and in the value of a clinical log book. In the theme of skills required for clinical dental practice, a statistically significant difference was seen between teacher and student groups in the value of a critical appreciation of evidence-based practice as one of the skills. The study overall indicates that the dental clinical learning environment supports close perceptual conformity between students and clinical teachers in regard to what each group considers to be "good practice" in clinical teaching. The findings of this study indicate that some techniques that have been advocated to enhance clinical learning, such as evidence-based teaching methods, require further investigation. PMID- 16352775 TI - Self-assessed dental status, oral behavior, DMF, and dental anxiety. AB - The purpose of this study was to evaluate the relation between self-assessed dental status, oral behavior, dental anxiety, and DMF scores. The study was conducted among young army personnel (eighteen to twenty-one years old) who arrived at a single military dental clinic for dental screening. A questionnaire was used to collect the data anonymously. The questionnaire included self assessment of dental status and behavior, as well as the Dental Anxiety Scale (DAS). Dental caries status was evaluated using the World Health Organization (WHO) caries diagnostic criteria for decayed, missing, and filled teeth (DMFT). A total of 393 young adults were examined (98.3 percent response rate). Approximately one-third brushed their teeth once a day or not at all. Of the participants, 10.7 percent assessed their dental status as "Poor" and 46.3 percent as "Fair." Furthermore, 25.5 percent assessed their dental treatment needs as high. The average DAS score was 7.14. The average DMF score was 6.2. There was a correlation between self-reported tooth status, participants' assessment of their dental treatment needs, and DMF score (p<0.0001). Anxious participants assessed their dental treatment needs as higher (p=0.05). The dental practitioner could ask patients to assess their dental status prior to treatment or appointment. These findings will help the practitioner to accomplish more effective time and patient management. PMID- 16352776 TI - Ziprasidone- and lithium-induced neuroleptic malignant syndrome. AB - OBJECTIVE: To report a case of ziprasidone- and lithium-induced neuroleptic malignant syndrome (NMS). CASE SUMMARY: A 47-year-old white male with a history of schizoaffective disorder was admitted to the hospital due to an exacerbation of severe mania. He had been taking lithium 450 mg twice daily and divalproex sodium 750 mg/day. On hospital day 2, ziprasidone 80 mg twice daily was added, and as-needed doses of intramuscular ziprasidone 20 mg and lorazepam 2 mg were used for agitation. On day 6, the patient developed hyperthermia (39.4 degrees C), elevated creatine kinase 26,000 units/L and white blood cell (WBC) count (20.7 x 10(3)/microL), myoglobinuria, hypotension (68/40 mm Hg), altered mental status, and tachypnea (28 breaths/min). This case is notable for the absence of muscle rigidity, which presents in greater than 90% of patients with NMS taking traditional antipsychotics. DISCUSSION: This case of ziprasidone- and lithium induced NMS is of probable cause, as determined by the Naranjo probability scale. The patient presented with symptoms consistent with NMS 4 days after initiation of ziprasidone and lithium. The majority of NMS cases present with the core features of hyperthermia, muscle rigidity, and elevated CK levels. Other frequently seen symptoms include altered mental status, tachypnea, tachycardia, elevated WBC count, hypotension, diaphoresis, and myoglobinuria. Our patient presented with 2 of the core symptoms, but did not develop muscle rigidity at any time. NMS criteria include muscle rigidity as one of the major presenting symptoms. Recent literature suggests that perhaps NMS due to novel antipsychotics presents with less muscle rigidity than is seen with traditional agents due to their lower affinity for the dopamine D2 receptor. CONCLUSIONS: This case illustrates that NMS due to the novel antipsychotic ziprasidone may present with many of the core symptoms of the syndrome, but possibly less muscle rigidity than is seen with traditional agents. PMID- 16352777 TI - Probable loop diuretic-induced pancreatitis in a sulfonamide-allergic patient. AB - OBJECTIVE: To report the case of a patient with a prior allergy to a sulfonamide antibiotic who subsequently developed the same reaction when administered various loop diuretics. CASE SUMMARY: A 57-year-old female with cardiomyopathy and "sulfa" (trimethoprim/sulfamethoxazole) allergy documented as pancreatitis presented with symptoms consistent with pancreatitis after use of furosemide. She subsequently developed similar symptoms after multiple rechallenges with various loop diuretics including furosemide, bumetanide, and torsemide. The patient was placed on ethacrynic acid until she was desensitized to furosemide. She had been receiving oral furosemide for 5 months at the time of this report, with no complications. According to the Naranjo probability scale, this reaction was probable. DISCUSSION: Reactions associated with arylamine sulfonamide-containing antibiotics have been commonly reported; however, cross-reactions with non arylamine sulfonamide-containing medications have been rare. The time delay by which symptoms of pancreatitis presented following administration of loop diuretics suggests an immunologic pathway. In addition, while cases of loop diuretic-induced pancreatitis, including furosemide, have been published, the allergic manifestations with both sulfonamide antibiotics and non-antibiotics in our patient suggest possible cross-reactivity between these 2 drug classes. CONCLUSIONS: The mechanism by which loop diuretics induce pancreatitis appears to be via an immunologic pathway. While the true correlation remains unknown, allergic cross-reactivity may occur between sulfonamide antibiotics and non antibiotics, such as loop diuretics. Torsemide appears to also be a part of a long list of agents that can cause pancreatitis. PMID- 16352778 TI - Assessment of compliance with lipid guidelines in an academic medical center. AB - BACKGROUND: The importance of achieving a low-density lipoprotein cholesterol (LDL-C) level less than 100 mg/dL in patients with coronary artery disease (CAD) or cerebrovascular disease (CVD) is well established. Emerging evidence supports the recognition and management of secondary lipid goals, high-density lipoprotein cholesterol (HDL-C) level greater than 40 mg/dL, and triglyceride level less than 150 mg/dL. OBJECTIVE: To evaluate whether inpatient services within an academic setting were achieving/addressing primary and secondary lipid goals in patients with established CAD or CVD. METHODS: Patients with a discharge diagnosis of acute myocardial infarction, myocardial revascularization procedures, and/or ischemic stroke were identified. A retrospective chart review was done to assess adherence to the American Heart Association (AHA)/American College of Cardiology (ACC) guidelines for lipid management. RESULTS: On average, 63% of patients with CAD or CVD had a lipid panel assessed during their hospitalization. Of the patients who had a fasting lipid panel checked, only 40% (72/178) had an LDL-C level less than 100 mg/dL. Of those patients, only 31% (22) also had an HDL-C level greater than 40 mg/dL. Even fewer patients (24%; 17) met both primary and secondary goals. Of the 287 patients included in the study, 69% (199) were prescribed a statin, 3% (9) a fibrate, and 3% (8) niacin on discharge. CONCLUSIONS: Few patients with CAD or CVD met the AHA/ACC goals for lipid management, yet a significant number were not prescribed appropriate lipid lowering therapy at discharge. This finding strongly suggests that more awareness in this area is needed. PMID- 16352779 TI - No more than necessary: safety and efficacy of low-dose promethazine. AB - BACKGROUND: Limitations in antiemetic options have resulted in increased use of intravenous promethazine. However, this drug has significant sedative effects with its standard dosage of 25 mg, especially when used in conjunction with narcotic analgesics. While studies have revealed the bioavailability of enteric promethazine to be 25%, current dosing references suggest identical dosing regardless of the route of administration. OBJECTIVE: To compare the antiemetic efficacy and sedative effects of low-dose intravenous promethazine (6.25 or 12.5 mg) with intravenous ondansetron 4 mg. METHODS: We assessed inpatients with noncritical conditions at Anne Arundel Medical Center who were treated for nausea or vomiting from any cause except chemotherapy or pregnancy. Forty-six patients received low-dose promethazine and 41 received ondansetron. Statistical analysis was carried out for significant differences in efficacy and sedation. RESULTS: For patients who received intravenous promethazine 6.25 or 12.5 mg, nausea and vomiting were relieved at one hour in 74% and 68%, respectively, compared with 59% for intravenous ondansetron 4 mg. Results at 3 hours were 67% and 80% for promethazine and 71% for ondansetron. Median sedation scores at one hour were equal at 3 for promethazine and ondansetron (4 = fully awake); at 3 hours, the median scores were 4 and 3.5, respectively. There were no statistically significant differences among any of these results. CONCLUSIONS: Low-dose (6.25 mg) intravenous promethazine relieves nausea and vomiting as effectively as intravenous ondansetron 4 mg. PMID- 16352780 TI - The cover. Children at play in Mantua, Utah. PMID- 16352781 TI - A piece of my mind. Fourth floor. PMID- 16352782 TI - Researchers discover a range of factors undermine sperm quality, male fertility. PMID- 16352783 TI - Ovarian cancer inspires art exhibit. PMID- 16352784 TI - Gene fusion found in prostate cancer. PMID- 16352785 TI - Risk factors for cardiovascular disease in women. PMID- 16352786 TI - Risk factors for cardiovascular disease in women. PMID- 16352787 TI - Routine vs selective invasive strategies in acute coronary syndromes. PMID- 16352788 TI - Routine vs selective invasive strategies in acute coronary syndromes. PMID- 16352789 TI - Circadian rhythms and reperfusion in patients with acute ST-segment elevation myocardial infarction. PMID- 16352790 TI - Cognitive therapy and preventing suicide attempts. PMID- 16352791 TI - Cognitive therapy and preventing suicide attempts. PMID- 16352792 TI - Dietary fiber intake and risk of colorectal cancer: a pooled analysis of prospective cohort studies. AB - CONTEXT: Inconsistent findings from observational studies have continued the controversy over the effects of dietary fiber on colorectal cancer. OBJECTIVE: To evaluate the association between dietary fiber intake and risk of colorectal cancer. DESIGN, SETTING, AND PARTICIPANTS: From 13 prospective cohort studies included in the Pooling Project of Prospective Studies of Diet and Cancer, 725,628 men and women were followed up for 6 to 20 years across studies. Study- and sex-specific relative risks (RRs) were estimated with the Cox proportional hazards model and were subsequently pooled using a random-effects model. MAIN OUTCOME MEASURE: Incident colorectal cancer. RESULTS: During 6 to 20 years of follow-up across studies, 8081 colorectal cancer cases were identified. For comparison of the highest vs lowest study- and sex-specific quintile of dietary fiber intake, a significant inverse association was found in the age-adjusted model (pooled RR = 0.84; 95% confidence interval [CI], 0.77-0.92). However, the association was attenuated and no longer statistically significant after adjusting for other risk factors (pooled multivariate RR = 0.94; 95% CI, 0.86 1.03). In categorical analyses compared with dietary fiber intake of 10 to <15 g/d, the pooled multivariate RR was 1.18 (95% CI, 1.05-1.31) for less than 10 g/d (11% of the overall study population); and RR, 1.00 (95% CI, 0.85-1.17) for 30 or more g/d. Fiber intake from cereals, fruits, and vegetables was not associated with risk of colorectal cancer. The pooled multivariate RRs comparing the highest vs lowest study- and sex-specific quintile of dietary fiber intake were 1.00 (95% CI, 0.90-1.11) for colon cancer and 0.85 (95% CI, 0.72-1.01) for rectal cancer (P for common effects by tumor site = .07). CONCLUSIONS: In this large pooled analysis, dietary fiber intake was inversely associated with risk of colorectal cancer in age-adjusted analyses. However, after accounting for other dietary risk factors, high dietary fiber intake was not associated with a reduced risk of colorectal cancer. PMID- 16352793 TI - The long road to patient safety: a status report on patient safety systems. AB - CONTEXT: Since the Institute of Medicine (IOM) reports on medical errors and quality, national attention has focused on improving patient safety through changes in "systems" of care. These reports resulted in a new paradigm that, rather than centering on individual errors, focuses on the "systems" necessary to facilitate and enhance quality and protect patients. OBJECTIVES: To assess the status of hospital patient safety systems since the release of the IOM reports and to identify changes over time in 2 states that collaborated on a patient safety project funded by the Agency for Healthcare Research and Quality. DESIGN, SETTING, AND PARTICIPANTS: Survey of all acute care hospitals in Missouri and Utah at 2 points in time, in 2002 and 2004, using a 91-item comprehensive questionnaire (n = 126 for survey 1 and n = 128 for survey 2). To assess changes over time, we also studied the cohort of 107 hospitals that responded to both surveys. MAIN OUTCOME MEASURES: Responses to the 91-question survey as well as changes in responses to the survey questions over an 18-month period. Seven latent variables were constructed to represent the most important patient safety constructs studied: computerized physician order entry systems, computerized test results, and assessments of adverse events; specific patient safety policies; use of data in patient safety programs; drug storage, administration, and safety procedures; manner of handling adverse event/error reporting; prevention policies; and root cause analysis. For each hospital, the 7 latent variables were summed to give an overall measure of the patient safety status of the hospital. RESULTS: Development and implementation of patient safety systems is at best modest. Self-reported regression in patient safety systems was also found. While 74% of hospitals reported full implementation of a written patient safety plan, nearly 9% reported no plan. The area of surgery appears to have the greatest level of patient safety systems. Other areas, such as medications, with a long history of efforts in patient safety and error prevention, showed improvements, but the percentage of hospitals with various safety systems was already high at baseline for many systems. Some findings are surprising, given the overall trends; for example, while a substantial percentage of hospitals have medication safety systems, only 34.1% [corrected] reported full implementation at survey 2 of computerized physician order entry systems for medications, despite the growth of computer technology in general and in hospital billing systems in particular. CONCLUSIONS: The current status of hospital patient safety systems is not close to meeting IOM recommendations. Data are consistent with recent reports that patient safety system progress is slow and is a cause for great concern. Efforts for improvement must be accelerated. PMID- 16352794 TI - Prognostic value of serial B-type natriuretic peptide testing during follow-up of patients with unstable coronary artery disease. AB - CONTEXT: Elevated concentrations of B-type natriuretic peptide (BNP) at presentation in patients with acute coronary syndrome (ACS) are associated with long-term mortality. Few data exist regarding serial assessment of BNP levels during follow-up. OBJECTIVE: To determine whether concentrations of BNP at study entry (prior to hospital discharge for ACS) and at outpatient follow-up at 4 months and 12 months are associated with subsequent clinical outcomes. DESIGN, SETTING, AND PATIENTS: Prospective observational substudy of 4497 patients with non-ST-elevation or ST-elevation ACS who were enrolled in phase Z of the A to Z trial, which was conducted in 41 countries at 322 acute care hospitals between 1999 and 2003. MAIN OUTCOME MEASURE: Death from any cause or new onset of congestive heart failure (CHF) through 2 years. RESULTS: Levels of BNP were available in 4266 patients at study entry (prior to hospital discharge), 3618 patients at 4 months, and 2966 patients at 12 months. During follow-up there were 230 deaths and 163 incident cases of CHF. Adjusting for age, sex, index event, renal function, hypertension, prior heart failure, and diabetes, elevated levels of BNP (>80 pg/mL) were associated with subsequent death or new CHF when measured at study entry (111 [21%] vs 246 [7%]; adjusted hazard ratio [HR], 2.5; 95% confidence interval [CI], 2.0-3.3), at 4 months (34 [19%] vs 125 [4%]; adjusted HR, 3.9; 95% CI, 2.6-6.0), and at 12 months (19 [11%] vs 37 [1%]; adjusted HR, 4.7; 95% CI, 2.5-8.9). Patients with newly elevated levels of BNP at 4 months were at increased risk of death or new CHF (10 [15%] vs 105 [3%]); HR, 4.5; 95% CI, 2.3-8.6). Patients with elevated levels of BNP at study entry and with BNP levels lower than 80 pg/mL at 4 months tended to have only modestly increased risk (HR, 1.7; 95% CI, 1.0-2.9) compared with patients with BNP levels lower than 80 pg/mL at both visits. CONCLUSIONS: Serial determinations of BNP levels during outpatient follow-up after ACS predict the risk of death or new CHF. Changes in BNP levels over time are associated with long-term clinical outcomes and may provide a basis for enhanced clinical decision making in patients after onset of ACS.Clinical Trials Registration ClinicalTrials.gov Identifier: NCT00251576. PMID- 16352795 TI - Insulin, glucose, insulin resistance, and pancreatic cancer in male smokers. AB - CONTEXT: Obesity, diabetes mellitus, and glucose intolerance have been associated with increased pancreatic cancer risk; however, prediagnostic serum insulin concentration has not been evaluated as a predictor of this malignancy. OBJECTIVE: To investigate whether prediagnostic fasting glucose and insulin concentrations and insulin resistance are associated with subsequent incidence of exocrine pancreatic cancer in a cohort of male smokers. DESIGN, SETTING, AND PARTICIPANTS: A case-cohort prospective study within the Alpha-Tocopherol, Beta Carotene Cancer Prevention Study (1985-1988) cohort of 29,133 male Finnish smokers ages 50 to 69 years. The study included 400 randomly sampled subcohort control participants and 169 incident pancreatic cancer cases that occurred after the fifth year of follow-up. All participants were followed up through December 2001 (up to 16.7 years of follow-up). MAIN OUTCOME MEASURES: Incident exocrine pancreatic cancer identified from the Finnish Cancer Registry. RESULTS: After adjusting for age, smoking, and body mass index, higher baseline fasting serum concentrations of glucose, insulin, and insulin resistance were positively associated with pancreatic cancer. The presence of biochemically defined diabetes mellitus (glucose, > or =126 mg/dL [> or =6.99 mmol/L]) and insulin concentration in the highest vs lowest quartile both showed a significant 2-fold increased risk (hazard ratio [HR], 2.13; 95% confidence interval [CI], 1.04-4.35; and HR, 2.01; 95% CI, 1.03-3.93; respectively). There were significant interactions for all the biomarker exposures by follow-up time, such that the positive associations were stronger among the cases that occurred more than 10 years after baseline (highest vs lowest quartile: glucose, HR, 2.16; 95% CI, 1.05-4.42; P for trend = .02; insulin, HR, 2.90; 95% CI, 1.22-6.92; P for trend = .005; and insulin resistance, HR, 2.71; 95% CI, 1.19-6.18; P for trend = .006). CONCLUSIONS: These results support the hypothesis that exposure to higher insulin concentrations and insulin resistance predicts the risk of exocrine pancreatic cancer. PMID- 16352796 TI - Socioeconomic status in health research: one size does not fit all. AB - Problems with measuring socioeconomic status (SES)-frequently included in clinical and public health studies as a control variable and less frequently as the variable(s) of main interest-could affect research findings and conclusions, with implications for practice and policy. We critically examine standard SES measurement approaches, illustrating problems with examples from new analyses and the literature. For example, marked racial/ethnic differences in income at a given educational level and in wealth at a given income level raise questions about the socioeconomic comparability of individuals who are similar on education or income alone. Evidence also shows that conclusions about nonsocioeconomic causes of racial/ethnic differences in health may depend on the measure-eg, income, wealth, education, occupation, neighborhood socioeconomic characteristics, or past socioeconomic experiences-used to "control for SES," suggesting that findings from studies that have measured limited aspects of SES should be reassessed. We recommend an outcome- and social group-specific approach to SES measurement that involves (1) considering plausible explanatory pathways and mechanisms, (2) measuring as much relevant socioeconomic information as possible, (3) specifying the particular socioeconomic factors measured (rather than SES overall), and (4) systematically considering how potentially important unmeasured socioeconomic factors may affect conclusions. Better SES measures are needed in data sources, but improvements could be made by using existing information more thoughtfully and acknowledging its limitations. PMID- 16352797 TI - Ventilatory management of acute lung injury and acute respiratory distress syndrome. AB - CONTEXT: The acute lung injury and acute respiratory distress syndrome are critical illnesses associated with significant morbidity and mortality. Mechanical ventilation is the cornerstone of supportive therapy. However, despite several important advances, the optimal strategy for ventilation and adjunctive therapies for patients with acute lung injury and acute respiratory distress syndrome is still evolving. EVIDENCE ACQUISITION: To identify reports of invasive ventilatory and adjunctive therapies in adult patients with acute lung injury and acute respiratory distress syndrome, we performed a systematic English-language literature search of MEDLINE (1966-2005) using the Medical Subject Heading respiratory distress syndrome, adult, and related text words, with emphasis on randomized controlled trials and meta-analyses. EMBASE and the Cochrane Central Register of Controlled Trials were similarly searched. The search yielded 1357 potential articles of which 53 were relevant to the study objectives and considered in this review. EVIDENCE SYNTHESIS: There is strong evidence to support the use of volume- and pressure-limited lung-protective ventilation in adult patients with acute lung injury and acute respiratory distress syndrome. The benefit of increased levels of positive end-expiratory pressure and recruitment maneuvers is uncertain and is being further evaluated in ongoing trials. Existing randomized controlled trials of alternative ventilation modes, such as high-frequency oscillation and adjunctive therapies, including inhaled nitric oxide and prone positioning demonstrate no significant survival advantage. However, they may have a role as rescue therapy for patients with acute respiratory distress syndrome with refractory life-threatening hypoxemia. CONCLUSIONS: Volume- and pressure-limited ventilation strategies should be used in managing adult acute lung injury and acute respiratory distress syndrome patients. Further research is needed to identify barriers to widespread adoption of this strategy, as well as the role of alternative ventilation modes and adjunctive therapies. PMID- 16352798 TI - Electron-beam computed tomography for coronary calcium: a useful test to screen for coronary heart disease? PMID- 16352799 TI - The use of stories in clinical research and health policy. PMID- 16352800 TI - Dietary fiber and colorectal cancer: an ongoing saga. PMID- 16352801 TI - Creating a safer health care system: finding the constraint. PMID- 16352802 TI - JAMA patient page. Pancreatic cancer. PMID- 16352803 TI - Teaching resources. Imaging signal transduction in living cells with fluorescent proteins. AB - Until recently, studies in this field of signal transduction have involved the "what" and "when" of signaling. Who talks to whom and for how long? With the advent of genetically encoded fluorescent proteins, it has become possible to monitor signaling events in living cells in real time. This has added the dimension of "where" to the study of cellular signaling. This lecture, which is a part of "Cell Signaling Systems: A Course for Graduate Students," provides a survey of how green fluorescent protein (GFP)-tagged probes for signaling events have been used to elucidate new pathways, to describe the kinetics of signaling events at the single-cell level, and to reveal upon which subcellular compartments these events take place. Some of the findings confirm previous ones using biochemical techniques, and others have been surprising. Examples include those utilizing protein localization, relocalization, fluorescence recovery after photobleaching (FRAP), and fluorescence resonance energy transfer (FRET). The design of FRET probes is described. The detection of small guanosine triphosphatase (GTPase) signaling in living cells is used as an example to explore the creative and diverse ways investigators have developed to look at this system. PMID- 16352804 TI - Differentiation of naive cord-blood T cells into CD19-specific cytolytic effectors for posttransplantation adoptive immunotherapy. AB - Disease relapse is a barrier to achieving therapeutic success after unrelated umbilical cord-blood transplantation (UCBT) for B-lineage acute lymphoblastic leukemia (B-ALL). While adoptive transfer of donor-derived tumor-specific T cells is a conceptually attractive approach to eliminating residual disease after allogeneic hematopoietic stem cell transplantation, adoptive immunotherapy after UCBT is constrained by the difficulty of generating antigen-specific T cells from functionally naive umbilical cord-blood (UCB)-derived T cells. Therefore, to generate T cells that recognize B-ALL, we have developed a chimeric immunoreceptor to redirect the specificity of T cells for CD19, a B-lineage antigen, and expressed this transgene in UCB-derived T cells. An ex vivo process, which is compliant with current good manufacturing practice for T-cell trials, has been developed to genetically modify and numerically expand UCB-derived T cells into CD19-specific effector cells. These are capable of CD19-restricted cytokine production and cytolysis in vitro, as well as mediating regression of CD19+ tumor and being selectively eliminated in vivo. Moreover, time-lapse microscopy of the genetically modified T-cell clones revealed an ability to lyse CD19+ tumor cells specifically and repetitively. These data provide the rationale for infusing UCB-derived CD19-specific T cells after UCBT to reduce the incidence of CD19+ B-ALL relapse. PMID- 16352805 TI - Minimal molecular response in polycythemia vera patients treated with imatinib or interferon alpha. AB - Imatinib and recombinant interferon alpha (rIFNalpha) can induce remission in polycythemia vera (PV) patients, but gauging the depth of responses has not been possible due to lack of a specific disease marker. We found that patients undergoing imatinib (n = 14) or rIFNalpha (n = 7) therapy remained strongly positive for V617F JAK2, although there was a significant reduction in the median percentage of mutant alleles that correlated with hematologic response (P = .001). Furthermore, individuals who achieved complete hematologic remission had lower levels of V617F than those who did not (P = .001). Of 9 imatinib-treated cases for whom pretreatment samples were available, 7 with no or partial hematologic responses showed a marginal increase (median, 1.2-fold; range, 1.0 1.5) in the percentage of V617F alleles on treatment, whereas the 2 patients who achieved complete hematologic remission showed a 2- to 3-fold reduction. Our data indicate that, although PV patients may benefit from imatinib or rIFNalpha, molecular responses are relatively modest. PMID- 16352806 TI - Long-term engagement of CD6 and ALCAM is essential for T-cell proliferation induced by dendritic cells. AB - Interactions between T cells and antigen-presenting cells (APCs) are the first step in the induction of an adaptive immune response. Here, we show that CD6 and its ligand activated leukocyte cell adhesion molecule (ALCAM) are actively recruited to the antigen-induced dendritic cell (DC)-T-cell contact zone. Moreover, ALCAM-blocking antibodies interfere with DC-T-cell conjugate formation, demonstrating that CD6-ALCAM binding is essential for stable T-cell-APC contact. We now demonstrate that besides their role in establishing initial contacts, CD6 ALCAM interactions are also required during the proliferative phase of the T-cell response; the presence of CD6-blocking antibodies or recombinant ALCAM-Fc proteins results in a strong and sustained inhibition of T-cell proliferation. Furthermore, simultaneous crosslinking of CD6 and CD3 induces enhanced proliferation and transcriptional activity to a similar level as observed after CD3 and CD28 co-crosslinking, demonstrating that CD6 is an important costimulatory molecule. The stability of ALCAM-CD6 binding, which contrasts with transient homotypic ALCAM-ALCAM interactions, further supports the long-lasting effects observed on T-cell proliferation. Taken together, we demonstrate that CD6 and ALCAM form a key adhesive receptor-ligand pair that is not only involved in early DC-T-cell binding but also in sustaining DC-induced T-cell proliferation long after the initial contact has been established. PMID- 16352807 TI - Oral valganciclovir leads to higher exposure to ganciclovir than intravenous ganciclovir in patients following allogeneic stem cell transplantation. AB - Cytomegalovirus (CMV) infection is a major complication after allogeneic stem cell transplantation (SCT). Valganciclovir (V-GCV) is an oral prodrug hydrolyzed to the anti-CMV drug ganciclovir (GCV). A randomized, multicenter, crossover, open-label clinical trial compared exposure to GCV after V-GCV and intravenous GCV (IV-GCV) as preemptive therapy for CMV disease in SCT. The primary objective was to compare exposure to GCV in patients with CMV infection stratified for intestinal graft-versus-host disease (I-GVHD). Secondary objectives were the assessment of safety and efficacy. Patients without I-GVHD had a higher exposure to GCV after V-GCV when compared with IV-GCV (area under the concentration-time curve from drug administration to last observed concentration after 12 hours [AUC(0-12)] 53.8 +/- 17.97 microg/mL . h [mean +/- SD] vs 39.5 +/- 13.91; P < .001; ratio of V-GCV/IV-GCV was 1.4; 90% confidence interval [CI], 1.2-1.5). This was also true in patients with I-GVHD grades I-II (AUC(0-12) 52.9 +/- 21.75 vs 33.1 +/- 12.97 mug/mL . h; P = .018; ratio 1.6; 90% CI, 1.3-2.0). Absolute bioavailability of GCV after V-GCV was approximately 75% in individuals with or without I-GVHD grades I-II. No severe GCV-related toxicity was observed and efficacy and safety was comparable (84-day follow-up). This supports the use of V GCV in SCT, even in patients with I-GVHD grades I-II. Due to higher exposure after V-GCV compared with IV-GCV, patients should be monitored carefully for safety reasons. PMID- 16352808 TI - Donor CD4+ T and B cells in transplants induce chronic graft-versus-host disease with autoimmune manifestations. AB - Chronic graft-vs-host disease (GVHD) is a major cause of morbidity and mortality of long-term survivors of allogeneic hemato-poietic cell transplantation (HCT). Chronic GVHD can have features of an autoimmune collagen vascular disease with clinical manifestations similar to autoimmune scleroderma and systemic lupus erythematosus (SLE). However, the pathogenesis of chronic GVHD is poorly understood. It is unclear how autoreactive T and B cells are generated in chronic GVHD recipients. We have recently developed a new chronic GVHD model by transplantation of donor DBA/2 (H-2d) spleen cells into major histocompatibility complex (MHC)-matched but minor antigen-mismatched sublethally irradiated BALB/c (H-2d) recipients as well as athymic BALB/c(nu/nu) and adult-thymectomized BALB/c recipients. Both euthymic and athymic BALB/c recipients developed high levels of serum IgG autoantibodies, sclerodermatous skin damage, and glomerulonephritis. Disease induction required both donor CD25-CD4+ T and B cells in transplants. In contrast, donor CD25+CD4+ T regulatory (Treg) cells prevented the disease induction. These results indicate that host thymus is not required for induction of chronic GVHD and that quiescent autoreactive T and B cells in transplants from nonautoimmune donors may be activated and expanded to cause chronic GVHD with autoimmune manifestations in allogeneic recipients, and donor Treg cells can suppress this process. PMID- 16352809 TI - Arf6 plays an early role in platelet activation by collagen and convulxin. AB - Small GTPases play critical roles in hemostasis, though the roster of such molecules in platelets is not complete. In this study, we report the presence of Ras-related GTPases of the ADP-ribosylation factor (Arf) family. Platelets contain Arf1 or 3 and Arf6, with the latter being predominantly membrane associated. Using effector domain pull-down assays, we show, counter to other GTPases, that Arf6-GTP is present in resting platelets and decreases rapidly upon activation with collagen or convulxin. This decrease does not completely rely on secondary agonists (ADP and thromboxane A2) or require integrin signaling. The decrease in free Arf6-GTP temporally precedes activation of Rho family GTPases (RhoA, Cdc42, and Rac1). Using a membrane-permeant, myristoylated peptide, which mimics the N-terminus of Arf6, we show that the Arf6-GTP decrease is essential for collagen- and convulxin-induced aggregation, platelet adherence, and spreading on collagen-coated glass. Treatment with this peptide also affects the activation of Rho family GTPases, but has little effect on RalA and Rap1 or on agonist-induced calcium mobilization. These data show that Arf6 is a key element in activation through GPVI, and is required for activation of the Rho family GTPases and the subsequent cytoskeletal rearrangements needed for full platelet function. PMID- 16352810 TI - Single-agent arsenic trioxide in the treatment of newly diagnosed acute promyelocytic leukemia: durable remissions with minimal toxicity. AB - Arsenic trioxide, as a single agent, has proven efficacy in inducing molecular remission in patients with acute promyelocytic leukemia (APL). There is limited long-term outcome data with single-agent As2O3 in the management of newly diagnosed cases of APL. Between January 1998 to December 2004, 72 newly diagnosed cases of APL were treated with a regimen of single-agent As2O3 at our center. Complete hematologic remission was achieved in 86.1%. At a median follow-up of 25 months (range: 8-92 months), the 3-year Kaplan-Meier estimate of EFS, DFS, and OS was 74.87% +/- 5.6%, 87.21% +/- 4.93%, and 86.11% +/- 4.08%, respectively. Patients presenting with a white blood cell (WBC) count lower than 5 x 10(9)/L and a platelet count higher than 20 x 10(9)/L at diagnosis (n = 22 [30.6%]) have an excellent prognosis with this regimen (EFS, OS, and DFS of 100%). The toxicity profile, in the majority, was mild and reversible. After remission induction, this regimen was administered on an outpatient basis. Single-agent As2O3, as used in this series, in the management of newly diagnosed cases of APL, is associated with responses comparable with conventional chemotherapy regimens. Additionally, this regimen has minimal toxicity and can be administered on an outpatient basis after remission induction. PMID- 16352811 TI - Prospective phase 1/2 study of rituximab in childhood and adolescent chronic immune thrombocytopenic purpura. AB - We assessed safety and efficacy of rituximab in a prospective study of 36 patients, age 2.6 to 18.3 years, with severe chronic immune thrombocytopenic purpura (ITP). The primary outcome of sustained platelets above 50 x 10(9)/L (50,000/mm3) during 4 consecutive weeks, starting in weeks 9 to 12, was achieved by 11 of 36 patients (31%, confidence interval [CI], 16% to 48%). Median response time was 1 week (range, 1 to 7 weeks). Attainment of the primary outcome was not associated with age, prior pharmacologic responses, prior splenectomy, ITP duration, screening platelet count, refractoriness, or IgM reduction. First-dose, infusion-related toxicity was common (47%) despite premedication. Significant drug-related toxicities included third-dose hypotension (n = 1) and serum sickness (n = 2). Peripheral B cells were depleted in all subjects. IgM decreased 3.4% per week, but IgG did not significantly decrease. Rituximab was well tolerated, with manageable infusion-related side effects, but 6% of subjects developed serum sickness. Rituximab is beneficial for some pediatric patients with severe, chronic ITP. PMID- 16352812 TI - A phase 3 study of deferasirox (ICL670), a once-daily oral iron chelator, in patients with beta-thalassemia. AB - Deferasirox (ICL670) is a once-daily oral iron chelator developed for the treatment of chronic iron overload from blood transfusions. A comparative phase 3 trial was conducted to demonstrate the efficacy of deferasirox in regularly transfused patients with beta-thalassemia aged 2 years or older. Patients were randomized and received treatment with deferasirox (n = 296) or deferoxamine (n = 290), with dosing of each according to baseline liver iron concentration (LIC). The primary endpoint was maintenance or reduction of LIC; secondary endpoints included safety and tolerability, change in serum ferritin level, and net body iron balance. In both arms, patients with LIC values of 7 mg Fe/g dry weight (dw) or higher had significant and similar dose-dependent reductions in LIC and serum ferritin, and effects on net body iron balance. However, the primary endpoint was not met in the overall population, possibly due to the fact that proportionally lower doses of deferasirox relative to deferoxamine were administered to patients with LIC values less than 7 mg Fe/g dw. The most common adverse events included rash, gastrointestinal disturbances, and mild nonprogressive increases in serum creatinine. No agranulocytosis, arthropathy, or growth failure was associated with deferasirox administration. Deferasirox is a promising once-daily oral therapy for the treatment of transfusional iron overload. PMID- 16352813 TI - Critical role for the Ets transcription factor ELF-1 in the development of tumor angiogenesis. AB - The Ets transcription factors regulate a wide variety of biologic processes. Several members have been shown to play a role in regulating angiogenesis and vascular development. For example, the Ets factor ELF-1 is enriched in the developing vasculature of the embryo, where it regulates the expression of the Tie2 gene. We have determined that ELF-1 and Tie2 expression is also enriched in tumor blood vessels, and have identified a short peptide, 34 amino acids in length, corresponding to the terminal portion of the highly conserved ETS domain that potently blocks the function of ELF-1. A tailored ELF-1 blocking peptide, containing a 12-amino acid HIV-1 TAT protein, readily crosses the cell membrane and enters into the nucleus of endothelial cells, leading to a marked reduction in the expression of ELF-1 gene targets including Tie2 and endothelial nitric oxide synthase. Furthermore, the ELF-1 blocking peptide potently inhibits angiopoietin-1-mediated endothelial cell migration. Systemic administration of this peptide markedly attenuates B16 melanoma tumor growth and tumor-associated angiogenesis in nude mice. These results support the function of ELF-1 in the regulation of Tie2 gene expression during the development of tumor angiogenesis. PMID- 16352814 TI - ATRA resolves the differentiation block in t(15;17) acute myeloid leukemia by restoring PU.1 expression. AB - Tightly regulated expression of the transcription factor PU.1 is crucial for normal hematopoiesis. PU.1 knockdown mice develop acute myeloid leukemia (AML), and PU.1 mutations have been observed in some populations of patients with AML. Here we found that conditional expression of promyelocytic leukemia-retinoic acid receptor alpha (PML-RARA), the protein encoded by the t(15;17) translocation found in acute promyelocytic leukemia (APL), suppressed PU.1 expression, while treatment of APL cell lines and primary cells with all-trans retinoic acid (ATRA) restored PU.1 expression and induced neutrophil differentiation. ATRA-induced activation was mediated by a region in the PU.1 promoter to which CEBPB and OCT-1 binding were induced. Finally, conditional expression of PU.1 in human APL cells was sufficient to trigger neutrophil differentiation, whereas reduction of PU.1 by small interfering RNA (siRNA) blocked ATRA-induced neutrophil differentiation. This is the first report to show that PU.1 is suppressed in acute promyelocytic leukemia, and that ATRA restores PU.1 expression in cells harboring t(15;17). PMID- 16352815 TI - Randomized controlled trial of deferiprone or deferoxamine in beta-thalassemia major patients with asymptomatic myocardial siderosis. AB - Most deaths in beta-thalassemia major result from cardiac complications due to iron overload. Differential effects on myocardial siderosis may exist between different chelators. A randomized controlled trial was performed in 61 patients previously maintained on subcutaneous deferoxamine. The primary end point was the change in myocardial siderosis (myocardial T2(*)) over 1 year in patients maintained on subcutaneous deferoxamine or those switched to oral deferiprone monotherapy. The dose of deferiprone was 92 mg/kg/d and deferoxamine was 43 mg/kg for 5.7 d/wk. Compliance was 94% +/- 5.3% and 93% +/- 9.7% (P = .81), respectively. The improvement in myocardial T2(*) was significantly greater for deferiprone than deferoxamine (27% vs 13%; P = .023). Left ventricular ejection fraction increased significantly more in the deferiprone-treated group (3.1% vs 0.3% absolute units; P = .003). The changes in liver iron level (-0.93 mg/g dry weight vs -1.54 mg/g dry weight; P = .40) and serum ferritin level (-181 microg/L vs -466 microg/L; P = .16), respectively, were not significantly different between groups. The most frequent adverse events were transient gastrointestinal symptoms for deferiprone-treated patients and local reactions at the infusion site for deferoxamine. There were no episodes of agranulocytosis. Deferiprone monotherapy was significantly more effective than deferoxamine over 1 year in improving asymptomatic myocardial siderosis in beta-thalassemia major. PMID- 16352817 TI - Septum enlightenment: assembly of bacterial division proteins. PMID- 16352818 TI - Cooperativity between different nutrient receptors in germination of spores of Bacillus subtilis and reduction of this cooperativity by alterations in the GerB receptor. AB - The GerA nutrient receptor alone triggers germination of Bacillus subtilis spores with L-alanine or L-valine, and these germinations were stimulated by glucose and K+ plus the GerK nutrient receptor. The GerB nutrient receptor alone did not trigger spore germination with any nutrients but required glucose, fructose, and K+ (GFK) (termed cogerminants) plus GerK for triggering of germination with a number of L-amino acids. GerB and GerA also triggered spore germination cooperatively with l-asparagine, fructose, and K+ and either L-alanine or L valine. Two GerB variants (termed GerB*s) that were previously isolated by their ability to trigger spore germination in response to D-alanine do not respond to D alanine but respond to the same L-amino acids that stimulate germination via GerB plus GerK and GFK. GerB*s alone triggered spore germination with these L-amino acids, although GerK plus GFK stimulated the rates of these germinations. In contrast to l-alanine germination via GerA, spore germination via L-alanine and GerB or GerB* was not inhibited by D-alanine. These data support the following conclusions. (i) Interaction with GerK, glucose, and K+ somehow stimulates spore germination via GerA. (ii) GerB can bind and respond to L-amino acids, although normally either the binding site is inaccessible or its occupation is not sufficient to trigger spore germination. (iii) Interaction of GerB with GerK and GFK allows GerB to bind or respond to amino acids. (iv) In addition to spore germination due to the interaction between GerA and GerK, and GerB and GerK, GerB can interact with GerA to trigger spore germination in response to appropriate nutrients. (v) The amino acid sequence changes in GerB*s reduce these receptor variants' requirement for GerK and cogerminants in their response to L-amino acids. (vi) GerK binds glucose, GerB interacts with fructose in addition to L amino acids, and GerA interacts only with L-valine, L-alanine, and its analogs. (vii) The amino acid binding sites in GerA and GerB are different, even though both respond to L-alanine. These new conclusions are integrated into models for the signal transduction pathways that initiate spore germination. PMID- 16352819 TI - Characterization of the Pseudomonas putida mobile genetic element ISPpu10: an occupant of repetitive extragenic palindromic sequences. AB - We have characterized the Pseudomonas putida KT2440 insertion element ISPpu10. This insertion sequence encodes a transposase which exhibits homology to the transposases and specific recombinases of the Piv/Moov family, and no inverted repeats are present at the borders of its left and right ends, thus constituting a new member of the atypical IS110/IS492 family. ISPpu10 was found in at least seven identical loci in the KT2440 genome, and variants were identified having an extra insertion at distinct loci. ISPpu10 always appeared within the core of specific repetitive extragenic palindromic (REP) sequences TCGCGGGTAAACCCGCTCCTAC, exhibiting high target stringency. One intragenic target was found associated with the truncation of a GGDEF/EAL domain protein. After active in vitro transposition to a plasmid-borne target, a duplication of the CT (underlined above) at the junction as a consequence of the ISPpu10 insertion was experimentally demonstrated for the first time in the IS110/IS492 family. The same duplication was observed after transposition of ISPpu10 from a plasmid to the chromosome of P. putida DOT-T1E, an ISPpu10-free strain with REPs similar to those of strain KT2440. Plasmid ISPpu10-mediated rearrangements were observed in vivo under laboratory conditions and in the plant rhizosphere. PMID- 16352820 TI - Role of the novel OprD family of porins in nutrient uptake in Pseudomonas aeruginosa. AB - To circumvent the permeability barrier of its outer membrane, Pseudomonas aeruginosa has evolved a series of specific porins. These channels have binding sites for related classes of molecules that facilitate uptake under nutrient limited conditions. Here, we report on the identification of a 19-member family of porins similar to the basic-amino-acid-specific porin OprD. The members of this family fell into one of two phylogenetically distinct clusters, one bearing high similarity to OprD and the other bearing most similarity to the putative phenylacetic acid uptake porin PhaK of Pseudomonas putida. Analysis of the genome context, operon arrangement, and regulation of the PhaK-like porin OpdK indicated that it might be involved in vanillate uptake. This result was confirmed by demonstrating that an opdK mutant had a deficiency in the ability to grow on vanillate as a carbon source. To extrapolate these data to other paralogues within this family, the substrate specificities of 6 of the 17 remaining OprD homologues were inferred using an approach similar to that used with opdK. The specificities determined were as follows: OpdP, glycine-glutamate; OpdC, histidine; OpdB, proline; OpdT, tyrosine; OpdH, cis-aconitate; and OpdO, pyroglutamate. Thus, members of the OprD subfamily took up amino acids and related molecules, and those characterized members most similar to PhaK were responsible for the uptake of a diverse array of organic acids. These results imply that there is a functional basis for the phylogenetic clustering of these proteins and provide a framework for studying OprD homologues in other organisms. PMID- 16352821 TI - Identification of the lower baseplate protein as the antireceptor of the temperate lactococcal bacteriophages TP901-1 and Tuc2009. AB - The first step in the infection process of tailed phages is recognition and binding to the host receptor. This interaction is mediated by the phage antireceptor located in the distal tail structure. The temperate Lactococcus lactis phage TP901-1 belongs to the P335 species of the Siphoviridae family, which also includes the related phage Tuc2009. The distal tail structure of TP901 1 is well characterized and contains a double-disk baseplate and a central tail fiber. The structural tail proteins of TP901-1 and Tuc2009 are highly similar, but the phages have different host ranges and must therefore encode different antireceptors. In order to identify the antireceptors of TP901-1 and Tuc2009, a chimeric phage was generated in which the gene encoding the TP901-1 lower baseplate protein (bppL(TP901-1)) was exchanged with the analogous gene (orf53(2009)) of phage Tuc2009. The chimeric phage (TP901-1C) infected the Tuc2009 host strain efficiently and thus displayed an altered host range compared to TP901-1. Genomic analysis and sequencing verified that TP901-1C is a TP901-1 derivative containing the orf53(2009) gene in exchange for bppL(TP901-1); however, a new sequence in the late promoter region was also discovered. Protein analysis confirmed that TP901-1C contains ORF53(2009) and not the lower baseplate protein BppL(TP901-1), and it was concluded that BppL(TP901-1) and ORF53(2009) constitute antireceptor proteins of TP901-1 and Tuc2009, respectively. Electron micrographs revealed altered baseplate morphology of TP901-1C compared to that of the parental phage. PMID- 16352822 TI - Transcription of virulence factors in Staphylococcus aureus small-colony variants isolated from cystic fibrosis patients is influenced by SigB. AB - Staphylococcus aureus small-colony variants (SCVs) are believed to account in part for the persistence of S. aureus during chronic infections. Little is understood about the gene expression profile that may explain the phenotype and distinguish SCVs from prototype S. aureus strains. In this study, DNA array transcriptional profiles of clinical SCVs isolated from the airways of cystic fibrosis patients were obtained and compared to those obtained from a laboratory derived SCV strain (i.e., a respiratory-deficient hemB mutant) and prototype S. aureus strains. The genes commonly up-regulated in both hemB and clinical SCVs were found to be implicated in fermentation and glycolysis pathways. The well known virulence regulator agr was not activated in SCVs, and such strains had low levels of alpha-toxin (hla) gene expression. Clinical SCVs also had a transcriptional signature of their own. Of striking interest is that many genes, most of them under the positive control of the alternate sigma factor SigB, were specifically up-regulated and differed in that way from that seen in prototype S. aureus and the hemB mutant. Since SigB influences up-regulation of adhesin type genes while indirectly down-regulating exoproteins and toxins, we evaluated the internalization and persistence of SCVs in mammalian cells. Results showed that clinical SCVs persisted much more efficiently in cells than the hemB and prototype strains and that a sigB mutant was a poor persister. Thus, it appears that the agr locus plays a minor role in the regulation of the virulon of SCVs, unlike SigB, which may have a key role in intracellular persistence. PMID- 16352823 TI - RscA, a member of the MDR1 family of transporters, is repressed by CovR and required for growth of Streptococcus pyogenes under heat stress. AB - The ability of Streptococcus pyogenes (group A streptococcus [GAS]) to respond to changes in environmental conditions is essential for this gram-positive organism to successfully cause disease in its human host. The two-component system CovRS controls expression of about 15% of the GAS genome either directly or indirectly. In most operons studied, CovR acts as a repressor. We previously linked CovRS to the GAS stress response by showing that the sensor kinase CovS is required to inactivate the response regulator CovR so that GAS can grow under conditions of heat, acid, and salt stress. Here, we sought to identify CovR-repressed genes that are required for growth under stress. To do this, global transcription profiles were analyzed by microarrays following exposure to increased temperature (40 degrees C) and decreased pH (pH 6.0). The CovR regulon in an M type 6 strain of GAS was also examined by global transcriptional analysis. We identified a gene, rscA (regulated by stress and Cov), whose transcription was confirmed to be repressed by CovR and activated by heat and acid. RscA is a member of the MDR1 family of ABC transporters, and we found that it is required for growth of GAS at 40 degrees C but not at pH 6.0. Thus, for GAS to grow at 40 degrees C, CovR repression must be alleviated so that rscA can be transcribed to allow the production of this potential exporter. Possible explanations for the thermoprotective role of RscA in this pathogen are discussed. PMID- 16352824 TI - Identification and characterization of the genes involved in glycosylation pathways of mycobacterial glycopeptidolipid biosynthesis. AB - Glycopeptidolipids (GPLs) are major components present on the outer layers of the cell walls of several nontuberculous mycobacteria. GPLs are antigenic molecules and have variant oligosaccharides in mycobacteria such as Mycobacterium avium. In this study, we identified four genes (gtf1, gtf2, gtf3, and gtf4) in the genome of Mycobacterium smegmatis. These genes were independently inactivated by homologous recombination in M. smegmatis, and the structures of GPLs from each gene disruptant were analyzed. Thin-layer chromatography, gas chromatography-mass spectrometry, and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analyses revealed that the mutants Deltagtf1 and Deltagtf2 accumulated the fatty acyl-tetrapeptide core having O-methyl-rhamnose and 6-deoxy talose as sugar residues, respectively. The mutant Deltagtf4 possessed the same GPLs as the wild type, whereas the mutant Deltagtf3 lacked two minor GPLs, consisting of 3-O-methyl-rhamnose attached to O-methyl-rhamnose of the fatty acyl tetrapeptide core. These results indicate that the gtf1 and gtf2 genes are responsible for the early glycosylation steps of GPL biosynthesis and the gtf3 gene is involved in transferring a rhamnose residue not to 6-deoxy-talose but to an O-methyl-rhamnose residue. Moreover, a complementation experiment showed that M. avium gtfA and gtfB, which are deduced glycosyltransferase genes of GPL biosynthesis, restore complete GPL production in the mutants Deltagtf1 and Deltagtf2, respectively. Our findings propose that both M. smegmatis and M. avium have the common glycosylation pathway in the early steps of GPL biosynthesis but differ at the later stages. PMID- 16352825 TI - Isolation of RNA polymerase from Clostridium difficile and characterization of glutamate dehydrogenase and rRNA gene promoters in vitro and in vivo. AB - Clostridium difficile is the primary causative agent of antibiotic-associated diarrheal disease. To facilitate molecular genetic analysis of gene expression in this organism, methods were developed to study transcriptional regulation in vitro and in vivo. That is, C. difficile RNA polymerase was partially purified and shown to bind to and initiate transcription in vitro from bona fide C. difficile promoters for rRNA and glutamate dehydrogenase genes. In addition, primer extension analyses and a beta-glucuronidase reporter system were used to quantitate transcription from these promoters in vivo. With these tools in hand, it is now possible to characterize the behavior of any C. difficile gene in vivo and to study the regulation of its expression in detail. PMID- 16352826 TI - Genetic transplantation: Salmonella enterica serovar Typhimurium as a host to study sigma factor and anti-sigma factor interactions in genetically intractable systems. AB - In Salmonella enterica serovar Typhimurium, sigma(28) and anti-sigma factor FlgM are regulatory proteins crucial for flagellar biogenesis and motility. In this study, we used S. enterica serovar Typhimurium as an in vivo heterologous system to study sigma(28) and anti-sigma(28) interactions in organisms where genetic manipulation poses a significant challenge due to special growth requirements. The chromosomal copy of the S. enterica serovar Typhimurium sigma(28) structural gene fliA was exchanged with homologs of Aquifex aeolicus (an extreme thermophile) and Chlamydia trachomatis (an obligate intracellular pathogen) by targeted replacement of a tetRA element in the fliA gene location using lambda Red-mediated recombination. The S. enterica serovar Typhimurium hybrid strains showed sigma(28)-dependent gene expression, suggesting that sigma(28) activities from diverse species are preserved in the heterologous host system. A. aeolicus mutants defective for sigma(28)/FlgM interactions were also isolated in S. enterica serovar Typhimurium. These studies highlight a general strategy for analysis of protein function in species that are otherwise genetically intractable and a straightforward method of chromosome restructuring using lambda Red-mediated recombination. PMID- 16352827 TI - Mutations in the central cavity and periplasmic domain affect efflux activity of the resistance-nodulation-division pump EmhB from Pseudomonas fluorescens cLP6a. AB - The EmhABC efflux system in Pseudomonas fluorescens cLP6a is homologous to the multidrug and solvent efflux systems belonging to the resistance-nodulation division (RND) family and is responsible for polycyclic aromatic hydrocarbon transport, antibiotic resistance, and toluene efflux. To gain a better understanding of substrate transport in RND efflux pumps, the EmhB pump was subjected to mutational analysis. Mutagenesis of amino acids within the central cavity of the predicted three-dimensional structure of EmhB showed selective activity towards antibiotic substrates. An A384P/A385Y double mutant showed increased susceptibility toward rhodamine 6G compared to the wild type, and F386A and N99A single mutants showed increased susceptibility to dequalinium compared to the wild type. As well, the carboxylic acid side chain of D101, located in the central cavity region, was found to be essential for polycyclic aromatic hydrocarbon transport and resistance to all antibiotic substrates of EmhB. Phenylalanine residues located within the periplasmic pore domain were also targeted for mutagenesis, and the F325A and F281A mutations significantly impaired efflux activity for all EmhB substrates. One mutation (A206S) in the outer membrane protein docking domain increased antibiotic resistance and toluene tolerance, demonstrating the important role of this domain in transport activity. These data demonstrate the roles of the central cavity and periplasmic domains in the function of the RND efflux pump EmhB. PMID- 16352828 TI - Hfq is a regulator of F-plasmid TraJ and TraM synthesis in Escherichia coli. AB - The F plasmid of Escherichia coli allows horizontal DNA transfer between an F(+) donor cell and an F(-) recipient. Expression of the transfer genes is tightly controlled by a number of factors, including the following plasmid-encoded regulatory proteins: TraJ, the primary activator of the 33-kb tra operon, and the autoregulators TraM and TraY. Here, we demonstrate that the host RNA binding protein, Hfq, represses TraJ and TraM synthesis by destabilizing their respective mRNAs. Mating assays and immunoblot analyses for TraM and TraJ showed that transfer efficiency and protein levels increased in host cells containing a disruption in hfq compared to wild-type cells in stationary phase. The stability of transcripts containing a putative Hfq binding site located in the intergenic untranslated region between traM and traJ was increased in hfq mutant donor cells, suggesting that Hfq destabilizes these transcripts. Electrophoretic mobility shift assays demonstrated that Hfq specifically binds this region but not the antisense RNA, FinP, encoded on the opposite strand. Together, these findings indicate that Hfq regulates traM and traJ transcript stability by a mechanism separate from FinOP-mediated repression. PMID- 16352829 TI - The Pseudomonas aeruginosa ribbon-helix-helix DNA-binding protein AlgZ (AmrZ) controls twitching motility and biogenesis of type IV pili. AB - Pseudomonas aeruginosa is an opportunistic pathogen that is commonly found in water and soil. In order to colonize surfaces with low water content, P. aeruginosa utilizes a flagellum-independent form of locomotion called twitching motility, which is dependent upon the extension and retraction of type IV pili. This study demonstrates that AlgZ, previously identified as a DNA-binding protein absolutely required for transcription of the alginate biosynthetic operon, is required for twitching motility. AlgZ may be required for the biogenesis or function of type IV pili in twitching motility. Transmission electron microscopy analysis of an algZ deletion in nonmucoid PAO1 failed to detect surface pili. To examine expression and localization of PilA (the major pilin subunit), whole-cell extracts and cell surface pilin preparations were analyzed by Western blotting. While the PilA levels present in whole-cell extracts were similar for wild-type P. aeruginosa and P. aeruginosa with the algZ deletion, the amount of PilA on the surface of the cells was drastically reduced in the algZ mutant. Analysis of algZ and algD mutants indicates that the DNA-binding activity of AlgZ is essential for the regulation of twitching motility and that this is independent of the role of AlgZ in alginate expression. These data show that AlgZ DNA-binding activity is required for twitching motility independently of its role in alginate production and that this involves the surface localization of type IV pili. Given this new role in twitching motility, we propose that algZ (PA3385) be designated amrZ (alginate and motility regulator Z). PMID- 16352830 TI - Role of Salmonella enterica serovar Typhimurium two-component system PreA/PreB in modulating PmrA-regulated gene transcription. AB - The PmrA/PmrB two-component system encoded by the pmrCAB operon regulates the modification of Salmonella enterica serovar Typhimurium lipopolysaccharide leading to polymyxin B resistance. PmrA and PhoP are the only known activators of pmrCAB. A transposon mutagenesis screen for additional regulators of a pmrC::MudJ fusion led to the identification of a two-component system, termed PreA/PreB (pmrCAB regulators A and B), that controls the transcription of the pmrCAB operon in response to unknown signals. The initial observations indicated that insertions in, or a deletion of, the preB sensor, but not the preA response regulator, caused upregulation of pmrCAB. Interestingly, the expression of pmrCAB was not upregulated in a preAB mutant grown in LB broth, implicating PreA in the increased expression of pmrCAB in the preB strain. This was confirmed by overexpression of preA(+) in preAB or preB backgrounds, which resulted in significant upregulation or further upregulation of pmrCAB. No such effect was observed in any tested preB(+) backgrounds. Additionally, an ectopic construct expressing a preA[D51A] allele also failed to upregulate pmrC in any of the pre backgrounds tested, which implies that there is a need for phosphorylation in the activation of the target genes. The observed upregulation of pmrCAB occurred independently of the response regulators PmrA and PhoP. Although a preB mutation led to increased transcription of pmrCAB, this did not result in a measurable effect on polymyxin B resistance. Our genetic data support a model of regulation whereby, in response to unknown signals, the PreB sensor activates PreA, which in turn indirectly upregulates pmrCAB transcription. PMID- 16352831 TI - The Mycobacterium tuberculosis TrcR response regulator represses transcription of the intracellularly expressed Rv1057 gene, encoding a seven-bladed beta propeller. AB - The Mycobacterium tuberculosis TrcR response regulator binds and regulates its own promoter via an AT-rich sequence. Sequences within this AT-rich region determined to be important for TrcR binding were used to search the M. tuberculosis H37Rv genome to identify additional related TrcR binding sites. A similar AT-rich sequence was identified within the intergenic region located upstream of the Rv1057 gene. In the present work, we demonstrate that TrcR binds to a 69-bp AT-rich sequence within the Rv1057 intergenic region and generates specific contacts on the same side of the DNA helix. An M. tuberculosis trcRS deletion mutant, designated STS10, was constructed and used to determine that TrcR functions as a repressor of Rv1057 expression. Additionally, identification of the Rv1057 transcriptional start site suggests that a SigE-regulated promoter also mediates control of Rv1057 expression. Using selective capture of transcribed sequences (SCOTS) analysis as an evaluation of intracellular expression, Rv1057 was shown to be expressed during early M. tuberculosis growth in human macrophages, and the Rv1057 expression profile correlated with a gene that would be repressed by TrcR. Based on structural predictions, motif analyses, and molecular modeling, Rv1057 consists of a series of antiparallel beta-strands which adopt a beta-propeller fold, and it was determined to be the only seven bladed beta-propeller encoded in the M. tuberculosis genome. These results provide evidence of TrcR response regulator repression of the Rv1057 beta propeller gene that is expressed during growth of M. tuberculosis within human macrophages. PMID- 16352832 TI - Identification of the syr-syp box in the promoter regions of genes dedicated to syringomycin and syringopeptin production by Pseudomonas syringae pv. syringae B301D. AB - The phytotoxins syringopeptin and syringomycin are synthesized by nonribosomal peptide synthetases which are encoded by the syringomycin (syr) and syringopeptin (syp) genomic island of Pseudomonas syringae pv. syringae. Previous studies demonstrated that expression of the syr-syp genes was controlled by the salA-syrF regulatory pathway, which in turn was induced by plant signal molecules. In this study, the 132-kb syr-syp genomic island was found to be organized into five polycistronic operons along with eight individual genes based on reverse transcriptional PCR and bioinformatic analysis. The transcriptional start sites of the salA gene and operons III and IV were located 63, 75, and 104 bp upstream of the start codons of salA, syrP, and syrB1, respectively, using primer extension analysis. The predicted -10/-35 promoter region of operon IV was confirmed based on deletion and site-directed mutagenesis analyses of the syrB1::uidA reporter with beta-glucuronidase assays. A 20-bp conserved sequence (TGtCccgN(6)cggGaCA, termed the syr-syp box) with dyad symmetry around the -35 region was identified via computer analysis for the syr-syp genes/operons responsible for biosynthesis and secretion of syringomycin and syringopeptin. Expression of the syrB1::uidA fusion was decreased 59% when 6 bp was deleted from the 5' end of the syr-syp box in the promoter region of operon IV. These results demonstrate that the conserved promoter sequences of the syr-syp genes contribute to the coregulation of syringomycin and syringopeptin production. PMID- 16352834 TI - CcpC-dependent regulation of citB and lmo0847 in Listeria monocytogenes. AB - In Bacillus subtilis, the catabolite control protein C (CcpC) plays a critical role in regulating the genes encoding the enzymes of the tricarboxylic acid branch of the Krebs citric acid cycle. A gene encoding a potential CcpC homolog and two potential target genes were identified in the Listeria monocytogenes genome. In vitro gel mobility shift assays and DNase I footprinting experiments showed that L. monocytogenes CcpC (CcpC(Lm)) interacts with the promoter regions of citB(Lm) (the gene that is likely to encode aconitase) and lmo0847 (encoding a possible glutamine transporter) and that citrate is a specific inhibitor of this interaction. To study in vivo promoter activity, a new lacZ reporter system was developed. This system allows stable integration into the chromosome of a promoter region transcriptionally fused to a promoterless lacZ gene at a nonessential, ectopic locus. Analysis of strains carrying a citB(Lm)-lacZ or lmo0847-lacZ fusion revealed that CcpC(Lm) represses citB(Lm) and lmo0847 in media containing an excess of glucose and glutamine. In addition, regulation of citB(Lm) expression in rich medium was growth phase dependent; during exponential growth phase, expression was very low even in the absence of CcpC(Lm), but a higher level of citB(Lm) expression was induced in stationary phase, suggesting the involvement of another, as yet unidentified regulatory factor. PMID- 16352833 TI - Evidence for clonal evolution among highly polymorphic genes in methicillin resistant Staphylococcus aureus. AB - The evolution of Staphylococcus aureus has been described as predominantly clonal, based on evidence from seven housekeeping genes. We aimed to test if this was also true for more polymorphic genes. In a collection of 60 isolates including major European epidemic methicillin-resistant S. aureus (MRSA) and sporadic MRSA strains, we compared the partial gene sequences of seven housekeeping genes (arcC, aroE, glpF, gmk, pta, tpi, and yqiL), six core adhesion genes (present in all strains) (clfA, clfB, fnbA, map, sdrC, and spa), and four accessory adhesion genes (not present in all strains) (ebpS, fnbB, sdrD, and sdrE). Nucleotide diversity of adhesion genes was 2- to 10-fold higher than genes used for multilocus sequence typing. All genes showed evidence for purifying selection with a weakly reduced level among accessory adhesion genes. Among these highly variable genes, there was no evidence for a difference in molecular evolution between epidemic and sporadic strains. Gene trees constructed from concatenated sequences of housekeeping, core adhesion, and accessory adhesion genes were highly congruent, indicating clonality, despite some evidence for homologous exchange. Further evidence for clonality was found with an overall positive correlation of allelic and nucleotidic divergence for both seven housekeeping genes and six core adhesion genes. However, for small allelic differences that fit the demarcations of clonal complexes (CCs) there was no such correlation, suggesting that recombination occurred. Therefore, despite an overall clonal population structure, recombination between related isolates within CCs might have contributed to S. aureus evolution. PMID- 16352835 TI - The Pseudomonas aeruginosa lipid A deacylase: selection for expression and loss within the cystic fibrosis airway. AB - Lipopolysaccharide (LPS) is the major surface component of gram-negative bacteria, and a component of LPS, lipid A, is recognized by the innate immune system through the Toll-like receptor 4/MD-2 complex. Pseudomonas aeruginosa, an environmental gram-negative bacterium that opportunistically infects the respiratory tracts of patients with cystic fibrosis (CF), can synthesize various structures of lipid A. Lipid A from P. aeruginosa strains isolated from infants with CF has a specific structure that includes the removal of the 3 position 3-OH C10 fatty acid. Here we demonstrate increased expression of the P. aeruginosa lipid A 3-O-deacylase (PagL) in isolates from CF infants compared to that in environmental isolates. PagL activity was increased in environmental isolates by growth in medium limited for magnesium and decreased by growth at low temperature in laboratory-adapted strains of P. aeruginosa. P. aeruginosa PagL was shown to be an outer membrane protein by isopycnic density gradient centrifugation. Heterologous expression of P. aeruginosa pagL in Salmonella enterica serovar Typhimurium and Escherichia coli resulted in removal of the 3-OH C14 fatty acid from lipid A, indicating that P. aeruginosa PagL recognizes either 3-OH C10 or 3 OH C14. Finally, deacylated lipid A species were not observed in some clinical P. aeruginosa isolates from patients with severe pulmonary disease, suggesting that loss of PagL function can occur during long-term adaptation to the CF airway. PMID- 16352836 TI - Reconstruction and regulation of the central catabolic pathway in the thermophilic propionate-oxidizing syntroph Pelotomaculum thermopropionicum. AB - Obligate anaerobic bacteria fermenting volatile fatty acids in syntrophic association with methanogenic archaea share the intermediate bottleneck step in organic-matter decomposition. These organisms (called syntrophs) are biologically significant in terms of their growth at the thermodynamic limit and are considered to be the ideal model to address bioenergetic concepts. We conducted genomic and proteomic analyses of the thermophilic propionate-oxidizing syntroph Pelotomaculum thermopropionicum to obtain the genetic basis for its central catabolic pathway. Draft sequencing and subsequent targeted gap closing identified all genes necessary for reconstructing its propionate-oxidizing pathway (i.e., methylmalonyl coenzyme A pathway). Characteristics of this pathway include the following. (i) The initial two steps are linked to later steps via transferases. (ii) Each of the last three steps can be catalyzed by two different types of enzymes. It was also revealed that many genes for the propionate oxidizing pathway, except for those for propionate coenzyme A transferase and succinate dehydrogenase, were present in an operon-like cluster and accompanied by multiple promoter sequences and a putative gene for a transcriptional regulator. Proteomic analysis showed that enzymes in this pathway were up regulated when grown on propionate; of these enzymes, regulation of fumarase was the most stringent. We discuss this tendency of expression regulation based on the genetic organization of the open reading frame cluster. Results suggest that fumarase is the central metabolic switch controlling the metabolic flow and energy conservation in this syntroph. PMID- 16352837 TI - Transposon disruption of the complex I NADH oxidoreductase gene (snoD) in Staphylococcus aureus is associated with reduced susceptibility to the microbicidal activity of thrombin-induced platelet microbicidal protein 1. AB - The cationic molecule thrombin-induced platelet microbicidal protein 1 (tPMP-1) exerts potent activity against Staphylococcus aureus. We previously reported that a Tn551 S. aureus transposon mutant, ISP479R, and two bacteriophage back transductants, TxA and TxB, exhibit reduced in vitro susceptibility to tPMP-1 (tPMP-1(r)) compared to the parental strain, ISP479C (V. Dhawan, M. R. Yeaman, A. L. Cheung, E. Kim, P. M. Sullam, and A. S. Bayer, Infect. Immun. 65:3293-3299, 1997). In the current study, the genetic basis for tPMP-1(r) in these mutants was identified. GenBank homology searches using sequence corresponding to chromosomal DNA flanking Tn551 mutant strains showed that the fourth gene in the staphylococcal mnh operon (mnhABCDEFG) was insertionally inactivated. This operon was previously reported to encode a Na(+)/H(+) antiporter involved in pH tolerance and halotolerance. However, the capacity of ISP479R to grow at pH extremes and in high NaCl concentrations (1 to 3 M), coupled with its loss of transmembrane potential (DeltaPsi) during postexponential growth, suggested that the mnh gene products are not functioning as a secondary (i.e., passive) Na(+)/H(+) antiporter. Moreover, we identified protein homologies between mnhD and the nuo genes of Escherichia coli that encode components of a complex I NADH:ubiquinone oxidoreductase. Consistent with these data, exposures of tPMP-1 susceptible (tPMP-1(s)) parental strains (both clinical and laboratory derived) with either CCCP (a proton ionophore which collapses the proton motive force) or pieracidin A (a specific complex I enzyme inhibitor) significantly reduced tPMP induced killing to levels seen in the tPMP-1(r) mutants. To reflect the energization of the gene products encoded by the mnh operon, we have renamed the locus sno (S. aureus nuo orthologue). These novel findings indicate that disruption of a complex I enzyme locus can confer reduced in vitro susceptibility to tPMP-1 in S. aureus. PMID- 16352838 TI - The AidB component of the Escherichia coli adaptive response to alkylating agents is a flavin-containing, DNA-binding protein. AB - Upon exposure to alkylating agents, Escherichia coli increases expression of aidB along with three genes (ada, alkA, and alkB) that encode DNA repair proteins. In order to begin to identify the role of AidB in the cell, the protein was purified to homogeneity, shown to possess stoichiometric amounts of flavin adenine dinucleotide (FAD), and confirmed to have low levels of isovaleryl-coenzyme A (CoA) dehydrogenase activity. A homology model of an AidB homodimer was constructed based on the structure of a four-domain acyl-CoA oxidase. The predicted structure revealed a positively charged groove connecting the two active sites and a second canyon of positive charges in the C-terminal domain, both of which could potentially bind DNA. Three approaches were used to confirm that AidB binds to double-stranded DNA. On the basis of its ability to bind DNA and its possession of a redox-active flavin, AidB is predicted to catalyze the direct repair of alkylated DNA. PMID- 16352839 TI - Assembly of CS1 pili: the role of specific residues of the major pilin, CooA. AB - CS1 pili are important virulence factors of enterotoxigenic Escherichia coli strains associated with human diarrheal disease. They are the prototype for a family of pili that share extensive sequence similarity among their structural and assembly proteins. Only four linked genes, cooB, cooA, cooC, and cooD, are required to produce CS1 pili in E. coli K-12. To identify amino acids important for the function of the major pilin CooA, we used alanine substitution mutagenesis targeting conserved residues in the N and C termini of the protein. To test function, we examined cooA mutants for the ability to agglutinate bovine erythrocytes. Each hemagglutination-negative (HA(-)) cooA mutant was examined to identify its assembly pathway defect. CooA has been shown to be degraded in the absence of CooB (K. Voegele, H. Sakellaris, and J. R. Scott, Proc. Natl. Acad. Sci. USA 94:13257-13261, 1997). We found several HA(-) cooA mutants that produced no detectable CooA, suggesting that recognition by CooB is mediated by residues in both the N and C termini of CooA. In addition, we found that alanine substitution for some of the conserved residues in the C-terminal motif "AGxYxG(x(6))T," which is found in all subunits of this pilus family, had no effect on pilus formation. However, alanine substitution for some of the alternating hydrophobic residues within this motif prevented CooA from interacting with CooD, which serves as both the tip adhesin and nucleation protein for pilus formation. Thus, it appears that some, but not all, of the residues in both the N and C termini of CooA play a critical role in the intermolecular interactions of the major pilin with the other structural and assembly proteins. We anticipate that the results obtained here for CS1 pili in enterotoxigenic E. coli will help develop an understanding of the pilus assembly pathway used by CS1 family members in several important human pathogens. PMID- 16352840 TI - Cold-induced putative DEAD box RNA helicases CshA and CshB are essential for cold adaptation and interact with cold shock protein B in Bacillus subtilis. AB - The nucleic acid binding cold shock proteins (CSPs) and the cold-induced DEAD box RNA helicases have been proposed separately to act as RNA chaperones, but no experimental evidence has been reported on a direct cooperation. To investigate the possible interaction of the putative RNA helicases CshA and CshB and the CSPs from Bacillus subtilis during cold shock, we performed genetic as well as fluorescence resonance energy transfer (FRET) experiments. Both cshA and cshB genes could be deleted only in the presence of a cshB copy in trans, showing that the presence of one csh gene is essential for viability. The combined gene deletion of cshB and cspD resulted in a cold-sensitive phenotype that was not observed for either helicase or csp single mutants. In addition to the colocalization of the putative helicases CshA and CshB with CspB and the ribosomes in areas surrounding the nucleoid, we detected a strong FRET interaction in vivo between CshB and CspB that depended on active transcription. In contrast, a FRET interaction was not observed for CshB and the ribosomal protein L1. Therefore, we propose a model in which the putative cold-induced helicases and the CSPs work in conjunction to rescue misfolded mRNA molecules and maintain proper initiation of translation at low temperatures in B. subtilis. PMID- 16352841 TI - Genomic changes during chronic Helicobacter pylori infection. AB - The gastric pathogen Helicobacter pylori shows tremendous genetic variability within human populations, both in gene content and at the sequence level. We investigated how this variability arises by comparing the genome content of 21 closely related pairs of isolates taken from the same patient at different time points. The comparisons were performed by hybridization with whole-genome DNA microarrays. All loci where microarrays indicated a genomic change were sequenced to confirm the events. The number of genomic changes was compared to the number of homologous replacement events without loss or gain of genes that we had previously determined by multilocus sequence analysis and mathematical modeling based on the sequence data. Our analysis showed that the great majority of genetic changes were due to homologous recombination, with 1/650 events leading to a net gain or loss of genes. These results suggest that adaptation of H. pylori to the host individual may principally occur through sequence changes rather than loss or gain of genes. PMID- 16352842 TI - Divergence and mosaicism among virulent soil phages of the Burkholderia cepacia complex. AB - We have determined the genomic sequences of four virulent myophages, Bcep1, Bcep43, BcepB1A, and Bcep781, whose hosts are soil isolates of the Burkholderia cepacia complex. Despite temporal and spatial separations between initial isolations, three of the phages (Bcep1, Bcep43, and Bcep781, designated the Bcep781 group) exhibit 87% to 99% sequence identity to one another and most coding region differences are due to synonymous nucleotide substitutions, a hallmark of neutral genetic drift. Phage BcepB1A has a very different genome organization but is clearly a mosaic with respect to many of the genes of the Bcep781 group, as is a defective prophage element in Photorhabdus luminescens. Functions were assigned to 27 out of 71 predicted genes of Bcep1 despite extreme sequence divergence. Using a lambda repressor fusion technique, 10 Bcep781 encoded proteins were identified for their ability to support homotypic interactions. While head and tail morphogenesis genes have retained canonical gene order despite extreme sequence divergence, genes involved in DNA metabolism and host lysis are not organized as in other phages. This unusual genome arrangement may contribute to the ability of the Bcep781-like phages to maintain a unified genomic type. However, the Bcep781 group phages can also engage in lateral gene transfer events with otherwise unrelated phages, a process that contributes to the broader-scale genomic mosaicism prevalent among the tailed phages. PMID- 16352843 TI - Two residues in the anticodon recognition domain of the aspartyl-tRNA synthetase from Pseudomonas aeruginosa are individually implicated in the recognition of tRNAAsn. AB - In many organisms, the formation of asparaginyl-tRNA is not done by direct aminoacylation of tRNA(Asn) but by specific tRNA-dependent transamidation of aspartyl-tRNA(Asn). This transamidation pathway involves a nondiscriminating aspartyl-tRNA synthetase (AspRS) that charges both tRNA(Asp) and tRNA(Asn) with aspartic acid. Recently, it has been shown for the first time in an organism (Pseudomonas aeruginosa PAO1) that the transamidation pathway is the only route of synthesis of Asn-tRNA(Asn) but does not participate in Gln-tRNA(Gln) formation. P. aeruginosa PAO1 has a nondiscriminating AspRS. We report here the identification of two residues in the anticodon recognition domain (H31 and G83) which are implicated in the recognition of tRNA(Asn). Sequence comparisons of putative discriminating and nondiscriminating AspRSs (based on the presence or absence of the AdT operon and of AsnRS) revealed that bacterial nondiscriminating AspRSs possess a histidine at position 31 and usually a glycine at position 83, whereas discriminating AspRSs possess a leucine at position 31 and a residue other than a glycine at position 83. Mutagenesis of these residues of P. aeruginosa AspRS from histidine to leucine and from glycine to lysine increased the specificity of tRNA(Asp) charging over that of tRNA(Asn) by 3.5-fold and 4.2 fold, respectively. Thus, we show these residues to be determinants of the relaxed specificity of this nondiscriminating AspRS. Using available crystallographic data, we found that the H31 residue could interact with the central bases of the anticodons of the tRNA(Asp) and tRNA(Asn). Therefore, these two determinants of specificity of P. aeruginosa AspRS could be important for all bacterial AspRSs. PMID- 16352844 TI - Polymorphisms in the nine polymorphic membrane proteins of Chlamydia trachomatis across all serovars: evidence for serovar Da recombination and correlation with tissue tropism. AB - Chlamydia trachomatis is an intracellular bacterium responsible for ocular, respiratory, and sexually transmitted diseases. The genome contains a nine-member polymorphic membrane protein (Pmp) family unique to members of the order Chlamydiales. Genomic and molecular analyses were performed for the entire pmp gene family for the 18 reference serological variants (serovars) and genovariant Ja to identify specific gene and protein regions that differentiate chlamydial disease groups. The mean genetic distance among all serovars varied from 0.1% for pmpA to 7.0% for pmpF. Lymphogranuloma venereum (LGV) serovars were the most closely related for the pmp genes and were also the most divergent, compared to ocular and non-LGV urogenital disease groups. Phylogenetic reconstructions showed that for six of nine pmp genes (not pmpA, pmpD, or pmpE), the serovars clustered based on tissue tropism. The most globally successful serovars, E and F, clustered distantly from the urogenital group for five pmp genes. These pmp genes may confer a biologic advantage that may facilitate infection and transmission for E and F. Surprisingly, serovar Da clustered with the ocular group from pmpE to pmpI, which are located together in the chromosome, providing statistically significant evidence for intergenomic recombination and acquisition of a genetic composition that could hypothetically expand the host cell range of serovar Da. We also identified distinct domains for pmpE, pmpF, and pmpH where substitutions were concentrated and associated with a specific disease group. Thus, our data suggest a possible structural or functional role that may vary among pmp genes in promoting antigenic polymorphisms and/or diverse adhesions-receptors that may be involved in immune evasion and differential tissue tropism. PMID- 16352845 TI - Gene-specific random mutagenesis of Escherichia coli in vivo: isolation of temperature-sensitive mutations in the acyl carrier protein of fatty acid synthesis. AB - Acyl carrier proteins (ACPs) are very small acidic proteins that play a key role in fatty acid and complex lipid synthesis. Moreover, recent data indicate that the acyl carrier protein of Escherichia coli has a large protein interaction network that extends beyond lipid synthesis. Despite extensive efforts over many years, no temperature-sensitive mutants with mutations in the structural gene (acpP) that encodes ACP have been isolated. We report the isolation of three such mutants by a new approach that utilizes error-prone PCR mutagenesis, overlap extension PCR, and phage lambda Red-mediated homologous recombination and that should be generally applicable. These mutants plus other experiments demonstrate that ACP function is essential for the growth of E. coli. Each of the mutants was efficiently modified with the phosphopantetheinyl moiety essential for the function of ACP in lipid synthesis, and thus lack of function at the nonpermissive temperature cannot be attributed to a lack of prosthetic group attachment. All of the mutant proteins were largely stable at the nonpermissive temperature except the A68T/N73D mutant protein. Fatty acid synthesis in strains that carried the D38V or A68T/N73D mutations was inhibited upon a shift to the nonpermissive temperature and in the latter case declined to a small percentage of the rate of the wild-type strain. PMID- 16352846 TI - A little gene with big effects: a serT mutant is defective in flgM gene translation. AB - A conditional-lethal mutant was isolated as having a flagellar regulatory phenotype at 30 degrees C and being unable to grow at 42 degrees C. Chromosomal mapping localized the mutation to the serT gene, which encodes an essential serine tRNA species (tRNA((cmo)5UGA)(Ser)). DNA sequence analysis revealed the mutation to be a single base change in G:A at position 10 of the serT gene that lies within the D-stem of the essential tRNA((cmo)5)UGA(Ser) species. tRNA((cmo)5)UGA(Ser) recognizes UCA, UCG, and UCU codons, but UCU is also recognized by tRNA(GGA)(Ser) and UCG by tRNA(CGA)(Ser). No other tRNAs are known to read the UCA codon. Thus, the UCA codon is specifically recognized by tRNA((cmo)5)UGA(Ser). We show that the anti-sigma(28) activity of FlgM is defective in the serT mutant strain. The serT allele causes a 10-fold increase in sigma(28)-dependent fliC promoter transcription, indicating a defect in FlgM anti sigma(28) activity in the presence of the serT mutation. The flgM gene contains only one UCA codon. Changing the UCA of flgM to ACG reversed the effect of the serT allele. Implications for context effects in regulation of gene expression are discussed. PMID- 16352847 TI - Autoinducer 2 controls biofilm formation in Escherichia coli through a novel motility quorum-sensing regulator (MqsR, B3022). AB - The cross-species bacterial communication signal autoinducer 2 (AI-2), produced by the purified enzymes Pfs (nucleosidase) and LuxS (terminal synthase) from S adenosylhomocysteine, directly increased Escherichia coli biofilm mass 30-fold. Continuous-flow cells coupled with confocal microscopy corroborated these results by showing the addition of AI-2 significantly increased both biofilm mass and thickness and reduced the interstitial space between microcolonies. As expected, the addition of AI-2 to cells which lack the ability to transport AI-2 (lsr null mutant) failed to stimulate biofilm formation. Since the addition of AI-2 increased cell motility through enhanced transcription of five motility genes, we propose that AI-2 stimulates biofilm formation and alters its architecture by stimulating flagellar motion and motility. It was also found that the uncharacterized protein B3022 regulates this AI-2-mediated motility and biofilm phenotype through the two-component motility regulatory system QseBC. Deletion of b3022 abolished motility, which was restored by expressing b3022 in trans. Deletion of b3022 also decreased biofilm formation significantly, relative to the wild-type strain in three media (46 to 74%) in 96-well plates, as well as decreased biomass (8-fold) and substratum coverage (19-fold) in continuous-flow cells with minimal medium (growth rate not altered and biofilm restored by expressing b3022 in trans). Deleting b3022 changed the wild-type biofilm architecture from a thick (54-mum) complex structure to one that contained only a few microcolonies. B3022 positively regulates expression of qseBC, flhD, fliA, and motA, since deleting b3022 decreased their transcription by 61-, 25-, 2.4-, and 18-fold, respectively. Transcriptome analysis also revealed that B3022 induces crl (26-fold) and flhCD (8- to 27-fold). Adding AI-2 (6.4 muM) increased biofilm formation of wild-type K-12 MG1655 but not that of the isogenic b3022, qseBC, fliA, and motA mutants. Adding AI-2 also increased motA transcription for the wild-type strain but did not stimulate motA transcription for the b3022 and qseB mutants. Together, these results indicate AI-2 induces biofilm formation in E. coli through B3022, which then regulates QseBC and motility; hence, b3022 has been renamed the motility quorum-sensing regulator gene (the mqsR gene). PMID- 16352848 TI - Comparative and functional genomic analysis of prokaryotic nickel and cobalt uptake transporters: evidence for a novel group of ATP-binding cassette transporters. AB - The transition metals nickel and cobalt, essential components of many enzymes, are taken up by specific transport systems of several different types. We integrated in silico and in vivo methods for the analysis of various protein families containing both nickel and cobalt transport systems in prokaryotes. For functional annotation of genes, we used two comparative genomic approaches: identification of regulatory signals and analysis of the genomic positions of genes encoding candidate nickel/cobalt transporters. The nickel-responsive repressor NikR regulates many nickel uptake systems, though the NikR-binding signal is divergent in various taxonomic groups of bacteria and archaea. B(12) riboswitches regulate most of the candidate cobalt transporters in bacteria. The nickel/cobalt transporter genes are often colocalized with genes for nickel dependent or coenzyme B(12) biosynthesis enzymes. Nickel/cobalt transporters of different families, including the previously known NiCoT, UreH, and HupE/UreJ families of secondary systems and the NikABCDE ABC-type transporters, showed a mosaic distribution in prokaryotic genomes. In silico analyses identified CbiMNQO and NikMNQO as the most widespread groups of microbial transporters for cobalt and nickel ions. These unusual uptake systems contain an ABC protein (CbiO or NikO) but lack an extracytoplasmic solute-binding protein. Experimental analysis confirmed metal transport activity for three members of this family and demonstrated significant activity for a basic module (CbiMN) of the Salmonella enterica serovar Typhimurium transporter. PMID- 16352849 TI - Solution structure of a novel tryptophan-rich peptide with bidirectional antimicrobial activity. AB - Trp-rich antimicrobial peptides play important roles in the host innate defense mechanisms of many plants, insects, and mammals. A new type of Trp-rich peptide, Ac-KWRRWVRWI-NH(2), designated Pac-525, was found to possess improved activity against both gram-positive and -negative bacteria. We have determined that the solution structures of Pac-525 bound to membrane-mimetic sodium dodecyl sulfate (SDS) micelles. The SDS micelle-bound structure of Pac-525 adopts an alpha helical segment at residues Trp2, Arg3, and Arg4. The positively charged residues are clustered together to form a hydrophilic patch. The three hydrophobic residues Trp2, Val6, and Ile9 form a hydrophobic core. The surface electrostatic potential map indicates the three tryptophan indole rings are packed against the peptide backbone and form an amphipathic structure. Moreover, the reverse sequence of Pac-525, Ac-IWRVWRRWK-NH(2), designated Pac-525(rev), also demonstrates similar antimicrobial activity and structure in membrane-mimetic micelles and vesicles. A variety of biophysical and biochemical methods, including circular dichroism, fluorescence spectroscopy, and microcalorimetry, were used to show that Pac-525 interacted strongly with negatively charged phospholipid vesicles and induced efficient dye release from these vesicles, suggesting that the antimicrobial activity of Pac-525 may be due to interactions with bacterial membranes. PMID- 16352850 TI - SecA dimer cross-linked at its subunit interface is functional for protein translocation. AB - SecA facilitates protein transport across the eubacterial plasma membrane by its association with cargo proteins and the SecYEG translocon, followed by ATP-driven conformational changes that promote protein translocation in a stepwise manner. Whether SecA functions as a monomer or a dimer during this process has been the subject of considerable controversy. Here we utilize cysteine-directed mutagenesis along with the crystal structure of the SecA dimer to create a cross linked dimer at its subunit interface, which was normally active for in vitro protein translocation. PMID- 16352851 TI - Generation of DNA-free Escherichia coli cells by 2-aminopurine requires mismatch repair and nonmethylated DNA. AB - Undirected mismatch repair initiated by the incorporation of the base analog 2 aminopurine kills DNA-methylation-deficient Escherichia coli dam cells by DNA double-strand breakage. Subsequently, the chromosomal DNA is totally degraded, resulting in DNA-free cells. PMID- 16352852 TI - Disruption of sucA, which encodes the E1 subunit of alpha-ketoglutarate dehydrogenase, affects the survival of Nitrosomonas europaea in stationary phase. AB - Although Nitrosomonas europaea lacks measurable alpha-ketoglutarate dehydrogenase activity, the recent completion of the genome sequence revealed the presence of the genes encoding the enzyme. A knockout mutation was created in the sucA gene encoding the E1 subunit. Compared to wild-type cells, the mutant strain showed an accelerated loss of ammonia monooxygenase and hydroxylamine oxidoreductase activities upon entering stationary phase. In addition, unlike wild-type cells, the mutant strain showed a marked lag in the ability to resume growth in response to pH adjustments in late stationary phase. PMID- 16352854 TI - Fetal programming of hypertension. AB - Numerous epidemiological studies suggest an inverse relationship between low birth weight (LBW) and hypertension, an observation now supported by numerous animal studies. The mechanisms linking LBW and hypertension appear to be multifactorial and involve alterations in the normal regulatory systems and renal functions involved in the long-term control of arterial pressure. Recent studies using animal models of fetal programming suggest that programming during fetal life occurs in response to an adverse fetal environment and results in permanent adaptive responses that lead to structural and physiological alterations and the subsequent development of hypertension. This review summarizes the adaptive responses observed in the different models used to induce a suboptimal fetal environment and discusses insights into the mechanisms mediating the fetal programming of hypertension. PMID- 16352853 TI - Mutations in Flavobacterium johnsoniae secDF result in defects in gliding motility and chitin utilization. AB - secDF mutants of Flavobacterium johnsoniae were deficient in gliding motility and chitin utilization. Cells of the mutants had reduced levels of GldJ protein, which is required for both processes. SecDF is similar to Escherichia coli SecD and SecF, which are involved in protein secretion. PMID- 16352855 TI - Role of fibroblast growth factor-binding protein in the pathogenesis of HIV associated hemolytic uremic syndrome. AB - A characteristic finding of childhood HIV-associated hemolytic uremic syndrome (HIV-HUS) is the presence of endothelial injury and microcystic tubular dilation, leading to a rapid progression of the renal disease. We have previously shown that a secreted fibroblast growth factor-binding protein (FGF-BP) is upregulated in kidneys from children affected with HIV-HUS and HIV nephropathy. Here, we sought to determine the potential role of FGF-BP in the pathogenesis of HIV-HUS. By immunohistochemical and in situ hybridization studies, we observed FGF-BP protein and mRNA upregulation in regenerating renal tubular epithelial cells from kidneys of HIV-Tg26 mice with late-stage renal disease, that is, associated with the development of microcystic tubular dilatation and accumulation of FGF-2. Moreover, FGF-BP increased the FGF-2-dependent growth and survival of cultured primary human renal glomerular endothelial cells and enhanced FGF-2-induced MAPK/ERK2 activation, as well as the proliferation of immortalized GM7373 endothelial cells. We propose that HIV-Tg26 mice are a clinically relevant model system to study the role of FGF-BP in the pathogenesis of HIV-associated renal diseases. Furthermore, the upregulation of FGF-BP by regenerating renal tubular epithelial cells may provide a mechanism by which the regenerative and angiogenic activity of FGF-2 in renal capillaries can be modulated in children with HIV-HUS and other renal disease. PMID- 16352856 TI - Physiological and pathophysiological aspects of ceramide. AB - Activation of cells by receptor- and nonreceptor-mediated stimuli not only requires a change in the activity of signaling proteins but also requires a reorganization of the topology of the signalosom in the cell. The cell membrane contains distinct domains, rafts that serve the spatial organization of signaling molecules in the cell. Many receptors or stress stimuli transform rafts by the generation of ceramide. These stimuli activate the acid sphingomyelinase and induce a translocation of this enzyme onto the extracellular leaflet of the cell membrane. Surface acid sphingomyelinase generates ceramide that serves to fuse small rafts and to form large ceramide-enriched membrane platforms. These platforms cluster receptor molecules, recruit intracellular signaling molecules to aggregated receptors, and seem to exclude inhibitory signaling factors. Thus ceramide-enriched membrane platforms do not seem to be part of a specific signaling pathway but may facilitate and amplify the specific signaling elicited by the cognate stimulus. This general function may enable these membrane domains to be critically involved in the induction of apoptosis by death receptors and stress stimuli, bacterial and viral infections of mammalian cells, and the regulation of cardiovascular functions. PMID- 16352857 TI - From clinical insights to new therapies. PMID- 16352858 TI - Improvements in insulin resistance with weight loss, in contrast to rosiglitazone, are not associated with changes in plasma adiponectin or adiponectin multimeric complexes. AB - It has been suggested that changes in adiponectin levels may contribute to improved insulin sensitivity in insulin-resistant individuals both after weight loss and after treatment with thiazolidinedione compounds. If this is correct, then changes in total circulating adiponectin and/or distribution of its multimeric complexes should coincide with improvements in insulin sensitivity after both interventions. To address this issue, fasting adiponectin concentrations and distribution of adiponectin complexes were measured in plasma samples in 24 insulin-resistant, nondiabetic subjects before and after 3-4 mo of treatment with either rosiglitazone or caloric restriction. The degree of insulin resistance in each group of 12 subjects was equal at baseline and improved to a similar extent ( approximately 30%) after each therapy. Whereas total adiponectin levels increased by nearly threefold and the relative amount of several higher molecular weight adiponectin complexes increased significantly in the rosiglitazone treatment group, there were no discernible changes in adiponectin levels or in the distribution between high or low molecular weight complexes in the weight loss group. These data indicate that, although changes in total adiponectin and several specific adiponectin complexes paralleled improvements in insulin resistance in thiazolidinedione-treated subjects, neither circulating adiponectin concentrations nor multimeric complexes changed in association with enhanced insulin sensitivity after moderate weight loss in 12 insulin-resistant, obese individuals. PMID- 16352859 TI - Endothelin-an emerging role in proinflammatory pathways in brain. PMID- 16352860 TI - Virus-induced obesity. PMID- 16352861 TI - Special feature: cardiovascular-kidney interactions in health and disease. PMID- 16352863 TI - D5 dopamine receptor regulation of reactive oxygen species production, NADPH oxidase, and blood pressure. AB - Activation of D1-like receptors (D1 and/or D5) induces antioxidant responses; however, the mechanism(s) involved in their antioxidant actions are not known. We hypothesized that stimulation of the D5 receptor inhibits NADPH oxidase activity, and thus the production of reactive oxygen species (ROS). We investigated this issue in D5 receptor-deficient (D5-/-) and wild-type (D5+/+) mice. NADPH oxidase protein expression (gp91(phox), p47(phox), and Nox 4) and activity in kidney and brain, as well as plasma thiobarbituric acid-reactive substances (TBARS) were higher in D5-/- than in D5+/+ mice. Furthermore, apocynin, an NADPH oxidase inhibitor, normalized blood pressure, renal NADPH oxidase activity, and plasma TBARS in D5-/- mice. In HEK-293 cells that heterologously expressed human D5 receptor, its agonist fenoldopam decreased NADPH oxidase activity, expression of one of its subunits (gp91(phox)), and ROS production. The inhibitory effect of the D5 receptor activation on NADPH oxidase activity was independent of cAMP/PKA but was partially dependent on phospholipase D2. The ability of D5 receptor stimulation to decrease ROS production may explain, in part, the antihypertensive action of D5 receptor activation. PMID- 16352862 TI - Resistance to renal damage by chronic nitric oxide synthase inhibition in the Wistar-Furth rat. AB - Chronic nitric oxide synthase inhibition (NOSI) causes chronic kidney disease (CKD) in the Sprague Dawley (SD) rat. We previously showed that the Wistar-Furth (WF) rats are resistant to several models of CKD and maintain renal nitric oxide (NO) production compared with SD rats, whereas low-dose NOSI caused progression of CKD in WF rats. Here, we evaluate the impact of high-dose chronic NOSI in WF and SD rats, as well as intrarenal responses to an acute pressor dose of NOSI in the normal WF. Rats were given N(G)-nitro-l-arginine methyl ester (l-NAME) (150 and 300 mg/l for 6-10 wk) in the drinking water after an initial bolus tail vein injection. Both strains showed significant reductions in total NO production with chronic l-NAME. SD given 150 mg/l l-NAME for 6 wk developed proteinuria and renal injury, whereas WF rats receiving 150 mg/l l-NAME for 6-10 wk or 300 mg/l for 6 wk developed no proteinuria and minimal renal injury. Blood pressure was significantly elevated with chronic NOSI in both strains but was higher in the SD rat. There was little impact on renal nitric oxide synthase expression with l NAME, except that cortical endothelial nitric oxide synthase abundance increased in WF after 6 wk (150 mg/l). Micropuncture experiments with acute pressor NOSI resulted in similar increases in systemic blood pressure in SD and WF rats, whereas WF rats showed a much smaller increment in glomerular blood pressure compared with SD rats. In conclusion, WF rats do not develop renal injury after chronic NOSI at, or above, a dose that causes significant injury in the SD rat. This protection may be associated with protection from glomerular hypertension. PMID- 16352864 TI - Quantum dots to monitor RNAi delivery and improve gene silencing. AB - A critical issue in using RNA interference for identifying genotype/phenotype correlations is the uniformity of gene silencing within a cell population. Variations in transfection efficiency, delivery-induced cytotoxicity and 'off target' effects at high siRNA concentrations can confound the interpretation of functional studies. To address this problem, we have developed a novel method of monitoring siRNA delivery that combines unmodified siRNA with seminconductor quantum dots (QDs) as multi color biological probes. We co-transfected siRNA with QDs using standard transfection techniques, thereby leveraging the photostable fluorescent nanoparticles to track delivery of nucleic acid, sort cells by degree of transfection and purify homogenously-silenced subpopulations. Compared to alternative RNAi tracking methods (co-delivery of reporter plasmids and end labeling the siRNA), QDs exhibit superior photostability and tunable optical properties for an extensive selection of non-overlapping colors. Thus this simple, modular system can be extended toward multiplexed gene knockdown studies, as demonstrated in a two color proof-of-principle study with two biological targets. When the method was applied to investigate the functional role of T cadherin (T-cad) in cell-cell communication, a subpopulation of highly silenced cells obtained by QD labeling was required to observe significant downstream effects of gene knockdown. PMID- 16352865 TI - Molecular flexibility in ab initio drug docking to DNA: binding-site and binding mode transitions in all-atom Monte Carlo simulations. AB - The dynamics of biological processes depend on the structure and flexibility of the interacting molecules. In particular, the conformational diversity of DNA allows for large deformations upon binding. Drug-DNA interactions are of high pharmaceutical interest since the mode of action of anticancer, antiviral, antibacterial and other drugs is directly associated with their binding to DNA. A reliable prediction of drug-DNA binding at the atomic level by molecular docking methods provides the basis for the design of new drug compounds. Here, we propose a novel Monte Carlo (MC) algorithm for drug-DNA docking that accounts for the molecular flexibility of both constituents and samples the docking geometry without any prior binding-site selection. The binding of the antimalarial drug methylene blue at the DNA minor groove with a preference of binding to AT-rich over GC-rich base sequences is obtained in MC simulations in accordance with experimental data. In addition, the transition between two drug-DNA-binding modes, intercalation and minor-groove binding, has been achieved in dependence on the DNA base sequence. The reliable ab initio prediction of drug-DNA binding achieved by our new MC docking algorithm is an important step towards a realistic description of the structure and dynamics of molecular recognition in biological systems. PMID- 16352866 TI - Gene expression studies in isolated mitochondria: Solanum tuberosum rps10 is recognized by cognate potato but not by the transcription, splicing and editing machinery of wheat mitochondria. AB - The complex gene expression mechanisms that occur in plant mitochondria, such as RNA editing and splicing, are not yet well understood. RNA editing in higher plant mitochondria is a highly specific process which modifies mRNA sequences by C-to-U conversions. It has been suggested that in some cases this process is required for splicing. Here, we use an experimental model based on the introduction of DNA into isolated mitochondria by electroporation to study organellar gene expression events. Our aim was to compare processing and editing of potato small ribosomal protein 10 gene (rps10) transcripts in heterologous (wheat mitochondria) and homologous (potato mitochondria) contexts. rps10 is a suitable model because it contains a group II intron, is absent in wheat mitochondria but is actively expressed in potato mitochondria, where transcripts are spliced and undergo five C-to-U editing events. For this purpose, conditions for electroporating isolated potato mitochondria were established. rps10 was placed under the control of either potato or wheat cox2 promoters. We found that rps10 was only transcribed under the control of a cognate promoter. In wheat mitochondria, rps10 transcripts were neither spliced nor edited while they are correctly processed in potato mitochondria. Interestingly, a wheat editing site grafted into rps10 was not recognized by wheat mitochondria but was correctly edited in potato mitochondria. Taken together, these results suggest that editing might occur only when the transcripts are engaged in processing and that they would not be available to editing factors outside of a putative RNA maturation machinery complex. PMID- 16352867 TI - Genomic blueprint of Hahella chejuensis, a marine microbe producing an algicidal agent. AB - Harmful algal blooms, caused by rapid growth and accumulation of certain microalgae in the ocean, pose considerable impacts on marine environments, aquatic industries and even public health. Here, we present the 7.2-megabase genome of the marine bacterium Hahella chejuensis including genes responsible for the biosynthesis of a pigment which has the lytic activity against a red-tide dinoflagellate. H.chejuensis is the first sequenced species in the Oceanospiralles clade, and sequence analysis revealed its distant relationship to the Pseudomonas group. The genome was well equipped with genes for basic metabolic capabilities and contained a large number of genes involved in regulation or transport as well as with characteristics as a marine heterotroph. Sequence analysis also revealed a multitude of genes of functional equivalence or of possible foreign origin. Functions encoded in the genomic islands include biosynthesis of exopolysacchrides, toxins, polyketides or non-ribosomal peptides, iron utilization, motility, type III protein secretion and pigmentation. Molecular structure of the algicidal pigment, which was determined through LC-ESI MS/MS and NMR analyses, indicated that it is prodigiosin. In conclusion, our work provides new insights into mitigating algal blooms in addition to genetic make up, physiology, biotic interactions and biological roles in the community of a marine bacterium. PMID- 16352868 TI - Human embryonic stem cells. PMID- 16352869 TI - Development and comparison of five site-specific biosphere models for safety assessment of radioactive waste disposal. AB - This paper describes the development and application of site-specific biosphere models that might be used for assessment of potential exposures in the framework of performance assessment studies of nuclear waste disposals. Model development follows the Reference Biosphere Methodology that has been set up in the framework of the BIOMASS study. In this paper, the application is to real sites at five European locations for which environmental and agricultural conditions have been described and characterised. For each of the sites a biosphere model has been developed specifically assuming a release of radionuclides to waters that are used by humans, for example as drinking water for humans and cattle and as irrigation water. Among the ingestion pathways, the intakes of drinking water, cereals, leafy vegetables, potatoes, milk, beef and freshwater fish are included in all models. Annual individual doses were calculated, and uncertainties in the results were estimated by means of stochastic calculations. To enable a comparison, all results were normalised to an activity concentration in groundwater of 1 Bq m(-3) for each of the radionuclides considered ((36)Cl, (79)Se, (99)Tc, (129)I, (135)Cs, (226)Ra, (231)Pa, (230)Th, (237)Np, (239)Pu, and (238)U), i.e. those that are usually most relevant in performance assessment studies of nuclear waste disposals. Although the results do not give answers in absolute terms on potential future exposures, they indicate the spectrum of exposures that might occur in different environments and specify the interaction of environmental conditions, human habits and potential exposure. PMID- 16352870 TI - Dexamethasone alters vascular reactivity by enhancing COX-related vasodilatation in fetal ovine carotids. AB - Based on preliminary studies that contractile efficacy was altered in vertebral and basilar arteries from neonatal donors treated with postnatal glucocorticoids, we examined the hypothesis that postnatal dexamethasone (DEX), a glucocorticoid used for respiratory disease in neonates, can alter vascular reactivity. Using near-term fetal lamb carotids, we measured 5-hydroxytryptamine (5-HT) dose response relationship in DEX-treated and untreated arteries. We found that DEX incubation for 1 h had no effect on 5-HT sensitivity and agonist affinity but significantly reduced 5-HT contractile efficacy, a response that became even more pronounced after 4 h of DEX treatment. Coincubation of DEX-treated arteries with INDO for 4 h reversed this DEX-induced attenuation in 5-HT contractile efficacy, although DEX had no significant effects on cyclooxygenase (COX)-1 and COX-2 protein abundance. This data suggests that DEX alters vascular reactivity through a COX-related mechanism, with possible repercussions to neurological injury. PMID- 16352872 TI - Stress cardiomyopathy with apical thrombosis promptly diagnosed with cardiovascular MRI. PMID- 16352871 TI - On-line haemodiafiltration versus haemodialysis: stable haematocrit with less erythropoietin and improvement of other relevant blood parameters. AB - BACKGROUND: Controlled randomised studies to prove improved cardiovascular stability and improved anaemia management during on-line haemodiafiltration (oHDF) are scarce. METHODS: 70 patients were treated with both haemodialysis (HD) and oHDF in a cross-over design during 2 x 24 weeks at a dialysis dose of eKt/V> or =1.2. Patients randomised into group A started on HD and switched over to oHDF, whereas patients in group B began with oHDF and were treated with HD afterwards. Intradialytic morbid events (IME), such as symptomatic hypotension or muscle cramps, were noted in case of appearance. Blood parameters reflecting anaemic status, phosphate status, lipid metabolism, oxidative stress, and accumulation of advanced glycation end products were recorded either monthly or at the end of each study phase. RESULTS: The mean incidence of IME was 0.15 IME per treatment, and there was no statistical difference between oHDF and HD. A higher haematocrit (oHDF 31.5% vs. HD 30.5%, p < 0.01) at a lower erythropoietin dose (oHDF 4,913 vs. HD 5,492 IU/week, p = 0.02) was found during oHDF, when the sequence of HD and oHDF had not been taken into account. For the study groups, the results were less distinct: in group A, a higher haematocrit (HD 30.4% vs. oHDF 32.0%, p < 0.01) at a comparable erythropoietin dose (HD 5,421 vs. oHDF 5,187 IU/week, ns) was observed during oHDF, whereas in group B an identical haematocrit (oHDF 30.8% vs. HD 30.7%, ns) was achieved at a reduced erythropoietin dose (oHDF 4,622 vs. HD 5,568 IU/week, p < 0.01). During oHDF, lower levels of free and protein-bound pentosidine and of serum phosphate were found. CONCLUSION: In contrast to other studies, no benefit regarding cardiovascular stability for oHDF was found, but oHDF could well offer a potential benefit regarding anaemia correction, inflammation, oxidative stress, lipid profiles, and calcium-phosphate product. PMID- 16352873 TI - The ability of selected oral microorganisms to emit red fluorescence. AB - Some novel caries detection and excavation devices rely on the ability of bacteria to produce red fluorescing compounds. The aim of this study was to examine the ability of selected oral microorganisms to emit red fluorescence. Streptococcus mutans, S. oralis, S. salivarius, S. sobrinus, Lactobacillus fermentans, L. casei, L. rhamnosus, Actinomyces naeslundi, A. israelii, Prevotella intermedia, and Fusobacterium nucleatum were inoculated onto Columbia agar with haemin and vitamin K and incubated anaerobically for up to 7 days in the dark. The resulting bacterial colonies were excited using filtered xenon light (405 +/- 20 nm) and digitally photographed through a 530-nm high-pass filter. The red and green portions of the colony fluorescence were analyzed using a computer program and the red/green ratio was calculated. All colonies emitted both red and green fluorescence. The green outweighed the red portion for the following species (in descending order) S. oralis, S. salivarius, S. mutans, F. nucleatum and S. sobrinus. The red portion was higher for the following species (in descending order) P. intermedia, A. naeslundi, A. israelii, L. fermentans, L. rhamnosus and L. casei. With all the bacteria examined, one color portion generally outweighed the other, giving the visual impression of either red or green fluorescence. We conclude that red fluorescence is well suited to detection of the bacteria which cause dentin caries but it is not suitable as an indicator of the presence and activity of the streptococci involved in initial caries. PMID- 16352874 TI - Serological diversity demonstrable by a set of monoclonal antibodies to eight serotypes of the mutans streptococci. AB - A set of monoclonal antibodies were prepared by the conventional cell fusion of myeloma cells (SP2/0-Ag14) with spleen cells from BALB/c mice immunised with whole cells of a strain of mutans streptococci. Their specificities were examined against 35 reference strains of mutans streptococci, 34 reference strains of other oral streptococci and 8 reference strains of other microorganisms often inhabiting the oral cavity. Specificity was examined by enzyme immunoassay using whole cells. A total of 52 strains, consisting of 19 strains isolated in Japan, 19 strains isolated in Italy and 14 strains isolated in England, were characterised by conventional physiological and biochemical tests and then serotyped by the use of 8 monoclonal antibodies with different specificities. They were also confirmed by guanine-plus-cytosine contents of their nucleic acid and DNA-DNA hybridisation test. The results indicated that all monoclonal antibodies are useful for identification of 8 serotypes of the mutans streptococci responsible for dental caries. They also suggest the existence of more serological varieties among mutans species. PMID- 16352875 TI - A rapid and quantitative detection system for Streptococcus mutans in saliva using monoclonal antibodies. AB - A new semiquantitative enumeration system was developed to detect Streptococcus mutans in saliva. Using species-specific monoclonal antibodies, the system quickly detected salivary S. mutans in 30 min and classified the results into three levels. In this study, saliva samples collected from 28 young adults aged between 22 and 24 years were subjected to the monoclonal antibody-based detecting system and selective medium-based detecting methods. The results generated from the PCR-confirmed culture method indicated the mean salivary S. mutans counts at level 1, 2 and 3 were 5.7 x 10(4), 1.3 x 10(5) and 3.4 x 10(6) CFU/ml, respectively. The differences between level 3 and 1 and level 3 and 2 were statistically significant (one-way ANOVA; p < 0.01). The results generated from the system were consistent with the data generated from two culture-based commercial products. PMID- 16352876 TI - Influence of cranberry juice on glucan-mediated processes involved in Streptococcus mutans biofilm development. AB - Cranberry juice (CJ) has biological properties that may provide health benefits. In this study, we investigated the influence of CJ (pH 5.5) on several activities in vitro associated with the development of Streptococcus mutans UA159 biofilms. The ability of CJ to influence the adherence of S. mutans to either saliva- (sHA) or glucan-coated hydroxyapatite (gsHA), and to inhibit the glucan production by purified glucosyltransferases adsorbed to sHA was determined. For the adherence assays, we used both uncoated and saliva-coated bacterial cells. Furthermore, we examined whether CJ interferes with the viability, development, polysaccharide composition and acidogenicity of S. mutans biofilms. A solution containing equivalent amounts of glucose, fructose and organic acids at pH 5.5 was used as negative control. The adherence of S. mutans (uncoated and saliva-coated) to either sHA or gsHA treated with 25% CJ (v/v) was remarkably reduced (40-85% inhibition compared to control: p < 0.05), indicating that CJ effectively blocked the bacterial adherence to binding sites in salivary pellicle and in glucans. In contrast, when the bacterial cells alone were treated with CJ they adhered to the similar untreated surfaces. Cranberry juice (25%, v/v) also inhibited the activities of surface-adsorbed GTF B and C (70-80% inhibition compared to control, p < 0.05). The effect of CJ on the viability of microorganisms in biofilms was not significant. Biofilm formation and accumulation were significantly reduced by topical applications of 25% CJ (v/v) twice daily with 1 min exposures (p < 0.05). The biomass and insoluble glucan content of the biofilms in addition to its acidogenicity were significantly reduced by cranberry treatments (p < 0.05). Our data show that cranberry juice inhibited glucan mediated biofilm development and acid production, and holds promise as a natural product to prevent biofilm-related oral diseases. PMID- 16352877 TI - Effect of sucrose concentration on dental biofilm formed in situ and on enamel demineralization. AB - The relationship between sucrose concentration and cariogenic potential was studied in situ. Adult volunteers wore intraoral palatal appliances containing human dental enamel blocks, which were extraorally submitted 8 times a day for 14 days, to the treatments: deionized distilled water and sucrose solutions from 1 to 40%. The biofilm formed was analyzed with respect to acidogenicity and biochemical composition; enamel demineralization was evaluated by microhardness. The results showed that 1% sucrose is less cariogenic than 5% or higher concentrations, although sucrose solution at 40% was still able to increase the concentration of insoluble polysaccharide in the biofilm formed. The findings suggest that the threshold of sucrose solution concentration for the formation of a cariogenic biofilm is 5%, which provided the same cariogenic potential as that observed for 10 and 20% sucrose solution. PMID- 16352878 TI - Acid susceptibility at various depths of pH-cycled enamel and dentine specimens. AB - pH-cycling and in situ studies have shown that fluctuations in de /remineralisation conditions or in fluoride usage can lead to laminations inside enamel or dentine lesions. Layers with different mineral content are thought to reflect the history of fluoride administrations. Studying the dissolution properties of such lesions at various depths -- using bulk specimens -- is presumably hampered by limited diffusion of acids through the lesion pores. Therefore, in this study the acid susceptibility of enamel and dentine lesions and the underlying sound tissues was studied by exposing sections to acid buffers from the cut rather than from the external surface. Specimens were obtained from a previous study of the effects of high-fluoride (0, 1,000, 2,000, 3,000/5,000 ppm F) toothpastes on enamel and dentine de-/remineralisation. Sections were subjected to acid buffers for 3 and 7 days and the changes in mineral content were monitored by contact microradiography. For enamel lesions a significant difference in dissolution over depth was observed between the groups subjected to the different fluoride schemes. At 7 days a dose response was found between the different fluoride groups and the lesion parameters. In the no-fluoride group dissolution in the original lesion and the sound tissue were similar. All dentine lesions which had been treated with fluoride showed inhibition of dissolution, but the inhibition did not increase with higher fluoride concentrations. Deeper into the dentine tissue there was some protection, but it was not statistically significant. We conclude that penetration of fluoride through the lesion pores determines the dissolution pattern of a lesion at various depths. PMID- 16352879 TI - The effect of adjacent dentine blocks on the demineralisation and remineralisation of enamel in vitro. AB - The aim was to investigate interactions between enamel and dentine at low pH under conditions simulating those at the enamel-dentine junction. Sound enamel blocks were demineralised in acid-gel systems, at pH 4.6, either in isolation, next to one, or in the middle of two, abutting dentine blocks. The gels were initially infinitely undersaturated with respect to enamel. In a second study, enamel blocks containing pre-formed lesions were placed in acid-gel systems, at pH 5.0, either in isolation or next to dentine blocks. The systems were initially either partially or infinitely undersaturated. In the partially saturated systems, calcium and phosphate concentrations were representative of plaque fluid. In the first study, demineralisation of enamel next to one dentine block was reduced in inverse proportion to the distance from the dentine. Demineralisation of enamel between two dentine blocks was retarded markedly across the whole block. In the second study, in the partially saturated systems, enamel lesions next to dentine blocks remineralised, whereas those in isolation demineralised further. We suggest that diffusion of dissolved dentine mineral over the enamel in the infinitely undersaturated system was sufficient to reduce undersaturation, thus retarding demineralisation, and that in the partially saturated systems, dentine dissolution together with the added calcium phosphate caused remineralisation of enamel lesions. Fluoride released from dissolving dentine may have augmented these effects. Different rates of demineralisation in enamel and dentine, or enamel remineralisation with concurrent dentine demineralisation, enabled by differences in their solubilities, could help explain the progression of so-called 'hidden caries'. PMID- 16352880 TI - The effect of adding calcium lactate to xylitol chewing gum on remineralization of enamel lesions. AB - The purpose of the study was to determine whether adding calcium lactate to chewing gum containing xylitol enhances remineralization of enamel surfaces using an early caries lesion model. Enamel slabs were cut from human extracted sound teeth and artificial subsurface lesions created within each. Half the enamel slabs were used as controls and stored in a humidifier while half were mounted into oral appliances worn by 10 volunteers (22-27 years old, 2 males and 8 females) in a three-leg trial, during which they wore the appliance without chewing gum, chewed gum containing xylitol + calcium lactate or chewed gum containing only xylitol 4 times a day for 2 weeks. Calcium concentrations in the enamel surfaces of control and test slabs were measured by X-ray spectrometry and degrees of remineralization were calculated. The mean degree of remineralization was greater after chewing xylitol-Ca gum (0.46 +/- 0.10) than after no gum (0.16 +/- 0.14) or after chewing xylitol gum (0.33 +/- 0.10) (p < 0.01). In conclusion, chewing gum containing xylitol + calcium lactate could enhance remineralization of enamel surface compared to chewing gum containing only xylitol or no gum chewing. PMID- 16352881 TI - An in vitro study of affected dentin as a risk factor for the development of secondary caries. AB - The objective of the study was to investigate the association between the presence of residual caries (inner affected dentin) on the cavity walls of cavity preparations and the further development of secondary caries lesions. Two 2 x 5 x 2 mm cavities, one in the apical portion (A) and one in the cervical portion (C) of the root, were prepared on both the lingual (L) and buccal (B) surfaces of 18 extracted human roots. The apical cavities (AB and AL) were artificially demineralized for 30 min using a decalcifying solution, followed by staining with a caries detector dye and then excavation of irreversibly demineralized dentin, leaving behind a layer of inner carious dentin. The cervical cavities (CB and CL) remained intact with sound dentin on the cavity walls. All cavities were then restored with composite resin. Following restoration, lingual specimens (AL and CL) were completely covered by an acid-resistant varnish to prevent further demineralization. All specimens were then incubated in an in vitro microbial artificial mouth model for 3 days in order to develop secondary carious lesions. At the end of the study all specimens were processed for energy-dispersive X-ray analysis of Ca concentration adjacent to the border between dentin and restoration. Statistical analysis of Ca concentrations revealed that the presence of affected inner dentin does not increase the susceptibility to secondary caries. Therefore, it was concluded that conservative cavity preparations leaving behind affected dentin do not increase the risk of secondary caries development. PMID- 16352882 TI - Caries-preventive effect of a one-time application of composite resin and glass ionomer sealants after 5 years. AB - The aim of the present trial was to (1) compare the caries-preventive effect of glass ionomer sealants, placed according to the atraumatic restorative treatment (ART) procedure, with composite resin sealants over time and (2) investigate the caries-preventive effect after complete disappearance of sealant material. Forty six boys and 57 girls, mean age 7.8 years, were randomly divided into two treatment groups in a parallel-group study design. A light-polymerized composite resin sealant material and a high-viscosity glass ionomer were each placed in 180 fully erupted first molars in their respective treatment groups. Evaluation took place annually for 5 years by calibrated examiners. After 5 years, 86% composite resin and 88% glass ionomer sealants did not survive. Three categories of re exposure periods for caries development in pits and fissures after complete loss of sealants were distinguished: 0-1, 1-2 and 2-3 years. In the 2- to 3-year group, 13 and 3% of pits and fissures previously sealed with composite resin and glass ionomer, respectively, were diagnosed as having developed a dentine lesion. The relative risks (95% CI) of dentine lesion development in surfaces sealed with glass ionomer compared to those sealed with composite resin after 3, 4 and 5 years were 0.22 (0.06-0.82), 0.32 (0.14-0.73) and 0.28 (0.13-0.61), respectively. The relative risks of dentine lesion development in pits and fissures previously sealed with glass ionomer compared with composite resin over re-exposure periods of 1-2 and 2-3 years were 0.26 (0.14-0.48) and 0.25 (0.09-0.68), respectively. We conclude that the caries-preventive effect of high-viscosity glass ionomer sealants, placed using the ART procedure, was between 3.1 and 4.5 times higher than that of composite resin sealants after 3-5 years. Furthermore, high viscosity (ART) glass ionomer sealants appear to have a four times higher chance of preventing caries development in re-exposed pits and fissures of occlusal surfaces in first molars than light-cured composite resin sealant material over a 1- to 3-year period. A well-designed clinical trial using different types of oral health personnel should be implemented to confirm these initial results. PMID- 16352883 TI - In vitro evaluation of abrasion of eroded enamel by different manual, power and sonic toothbrushes. AB - This study aimed to evaluate the susceptibility of eroded enamel to brushing abrasion performed by manual, power or sonic toothbrushes. Bovine enamel samples were subjected to 5 cycles, each consisting of 5 min demineralisation, 15 min remineralisation and 10 min brushing in a machine. Toothbrushing with the activated electric devices was supplemented with 20 linear strokes/min. Furthermore, enamel specimens were brushed with 20 linear strokes/min or 80 linear strokes/min with the electric toothbrushes without their individual operating action. A manual brush was applied at 100, 20 or 80 linear strokes/min. Specimens of the control group were not brushed after demineralisation. Loss of enamel was determined by profilometry. For all groups, substrate loss for linear brushing treatment applying 20 or 80 strokes/min did not differ significantly from the control (4.97 +/- 1.49 microm). Three power toothbrushing treatments significantly increased abrasion compared to linear brushing treatment with 20 or 80 strokes/min in their inactivated condition. The results indicate that brushing treatment with power or sonic toothbrushes may lead to significantly higher loss of demineralised enamel compared to toothbrushing without power or sonic support. PMID- 16352884 TI - The prevalence and severity of fluorosis in children who received toothpaste containing either 440 or 1,450 ppm F from the age of 12 months in deprived and less deprived communities. AB - This study compared fluorosis in the upper central incisors of children from socially diverse backgrounds who had received either 440- or 1,450-ppm F toothpaste from 12 months of age. The children were resident in non-fluoridated districts in the north-west of England. They received either 440- or 1,450-ppm F toothpaste and advice regarding its use until the age of 5-6 years. Dental fluorosis (TF index) was assessed on digital images of dried teeth when the children (n = 1,268) were 8-10 years old. In the less deprived districts the prevalences of fluorosis (TF >or=0) for the 1,450- and 440-ppm F groups were 34.5 and 23.7% (p = 0.006). In the deprived districts the prevalences of fluorosis were 25.2 and 19.5% (p = 0.2). Overall the prevalences of TF >or=2 were 7 and 2.1% for the 1,450- and 440-ppm F groups and 2.2 and 0.2% for TF >or=3. These differences were statistically significant (p < 0.003). There was a strong association between the deprivation status of wards and fluorosis. Only 1 subject with a TF score of 3 was identified in the two most deprived quintiles of the Townsend score. It is concluded that careful targeting of programmes of this type to children living in high caries risk deprived communities carries only a small risk of aesthetically objectionable fluorosis (TF >2) whether low or high fluoride toothpastes are used. High fluoride (1,450 ppm F) toothpastes should not be provided on a community basis to very young children in less deprived communities. PMID- 16352885 TI - The effects of illumination and focal distance on light-induced fluorescence images in vitro. AB - When using quantitative light-induced fluorescence a number of factors can influence illumination level. The purpose of this study was to investigate, using a high-resolution camera and fibre-optic light source, the impact of illumination level and focal distance on common quantitative light-induced fluorescence outcomes. Twenty-four extracted teeth were examined using 6 illumination levels and 4 focal distances. Analysis was conducted using multiple linear regression models fitted to log DeltaQ, log DeltaF and log area with clustering of teeth and robust standard errors. Separate models were used for the different light and focal levels. The regression coefficients were significant for both DeltaQ and DeltaF but not area. Despite the significant regressions the actual effect was very small, and unlikely to confound clinical trial or practice results. PMID- 16352886 TI - Caries occurrence in rats after bleaching with 10% carbamide peroxide in vivo. PMID- 16352887 TI - Interventional endoscopy and laparoscopy in GI diseases. PMID- 16352888 TI - Endoscopic therapy for oesophageal cancer. AB - Transthoracic surgery for oesophageal cancer is associated with a high incidence of respiratory complications. Recent development of minimally invasive oesophagectomy by the use of video-assisted thoracoscopic approach may have a potential to minimize morbidity and mortality. While results from earlier series were equivocal, recent reports have shown an encouraging trend. This article serves to review the recent literature evidence in relation to the surgical approach, safety, efficacy and potential problems of such a highly complex minimally invasive operation. PMID- 16352889 TI - Laparoscopic-endoscopic rendezvous resection of upper gastrointestinal tumors. AB - BACKGROUND: Endoscopic and laparoscopic local resection of gastric tumors has increasingly been performed in recent years. This article describes the technical considerations and early results of laparoscopic-endoscopic rendezvous resection of gastric lesions. PATIENTS AND METHODS: Rendezvous resection was performed in 26 patients with submucosal gastric tumors (n = 22) and early gastric cancer (n = 4). Laparoscopic wedge resection (LWR) was performed in 16 patients with anterior wall tumors and laparoscopic intragastric resection (LIR) in 7 patients with posterior wall tumors. Conversion to open surgery was carried out in 3 cases. RESULTS: Operation times were 53 min (range 35-115) for LWR and 83 min (range 56 130) for LIR, respectively. In submucosal lesions the mean tumor size was 36 mm (range 16-47) and in early gastric cancer 17.3 mm (range 16-20). Rendezvous resection was performed with curative intent and clear resection margins in all patients without lymphatic or vessel permeation. Postoperative complications occurred in 2 patients. After a mean follow-up of 22.8 months (range 2-71), no local recurrence or metastatic disease and no tumor-related death were observed. CONCLUSIONS: When selected properly, the laparoscopic-endoscopic approach is considered to be curative and safe for resection of localized gastric tumors. PMID- 16352890 TI - Laparoscopic gastrectomy for cancer. AB - There are three procedures for the management of early gastric cancer (EGC): laparoscopic wedge resection (LWR), intragastric mucosal resection (IGMR), and laparoscopic gastrectomy. LWR or IGMR can be applied to treat EGC without the risk of lymph node metastasis. However, owing to the recent technical advances in endoscopic mucosal resection for EGC, the use of laparoscopic local resection for these lesions has gradually decreased. On the other hand, laparoscopic gastrectomy with lymph node dissection, such as laparoscopy-assisted distal gastrectomy, is widely accepted for the treatment of EGC with the risk of lymph node metastasis. To establish the acceptability of laparoscopic gastrectomy with D2 lymph node dissection against advanced gastric cancers, safe techniques and new instruments must be developed. The following advantages of laparoscopic surgery for the treatment of gastric cancer have been well demonstrated: clinical course after operation, pulmonary function, immune response. In the future, laparoscopic surgeons have to design and implement education and training systems for standard laparoscopic procedures, evaluate clinical outcomes by multicentric randomized control trial studies, and clarify the oncological aspects of laparoscopic surgery in basic studies. PMID- 16352891 TI - Current status of laparoscopic therapy of cholecystolithiasis and common bile duct stones. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) is the standard operation for gallstone disease. The aim of this review was to scrutinize the advantages and benefits of this minimal invasive technique compared to the conventional operation according to the available literature. Regarding the evidence-based medicine criteria, the current status of laparoscopy in the treatment of cholecystolithiasis, cholecystitis and common bile duct stones has been worked out. METHODS: A Medline, PubMed, Cochrane search. RESULTS: Ten randomized controlled trials (RCTs) are available comparing laparoscopic versus open cholecystectomy. The superiority of LC in less postoperative pain, shorter recovery and hospital stay is stated. Operation time was longer in the first years of LC. 3 RCTs deal with acute cholecystitis: one paper could not find any significant advantage of LC over conventional cholecystectomy, the other two found benefits in recovery, hospital stay and postoperative pain. The range of conversion is between 5 and 7% in elective cases and increases up to 27% for acute cholecystitis. With a rate of more than 90% in Europe, the standard procedure for common bile duct stones is 'therapeutic splitting' with endoscopy and retrograde cholangiopancreatography preoperatively followed by LC. Laparoscopic bile duct clearance is effective and safe in experienced hands, however, the only proven benefit is a slightly shorter hospital stay. CONCLUSION: The laparoscopic approach is preferred in elective cholecystectomy and acute cholecystitis. The minimal invasive technique has proven to be effective, gentle and safe. The main benefits are evident within the first postoperative days. PMID- 16352892 TI - Current status of laparoscopic therapy of colorectal cancer. AB - Laparoscopic colorectal resections offer several benefits postoperatively, including minimal impairment of gastrointestinal and pulmonary function, less immunosuppression, shorter hospital stay and improved reconvalescence. Since the introduction of laparoscopic surgery for the therapy of curable colorectal cancer, some concern was voiced in terms of oncologic radicality, the issue of port-site metastases and tumor cell distribution. However, the clinical reality has demonstrated that oncologic radicality is equivalent to open surgery, and the incidence of port-site metastases is not increased when compared to wound recurrence at the laparotomy site. Focusing on colon and rectum, various indications of laparoscopic-endoscopic 'rendezvous' procedures exist including laparoscopic-assisted endoscopic transluminal resection, endoscopic-assisted wedge or anatomical resections, and, finally, intraoperative tumor location by colonoscopy to achieve oncologic resection margins in laparoscopic curative resections. In terms of colorectal curative resections, long-term results provide level I evidence that laparoscopic surgery for colon cancer is oncologically adequate and can be performed with equivalent morbidity and mortality rates when compared to conventional surgery. In terms of rectal cancer, no level I evidence is available. However, short-term data from experienced centers do not report inferior oncologic outcome particularly related to laparoscopic total mesorectal excision. PMID- 16352893 TI - Laparoscopic total mesorectal excision for rectal cancer surgery. AB - The introduction of total mesorectal excision in the early 1980s has improved local control and survival in patients with rectal cancer. Laparoscopic resections for colonic malignancies are gaining acceptance in the light of the recent evidence of oncologic adequacy from randomized clinical trials. Technical difficulties and the difference in the natural history of the disease have excluded rectal cancer from major clinical trials comparing laparoscopic to conventional rectal surgery. This review examined the feasibility, safety and oncologic outcomes of reported laparoscopic total mesorectal excision for surgical treatment of operable rectal cancer. PMID- 16352894 TI - Role and limitations of laparoscopic liver resection of colorectal metastases. AB - BACKGROUND: The current experience of laparoscopic liver resection is reviewed focusing on the role and limitations of resection of colorectal metastases. Surgical technique, outcome, and the main controversies regarding the procedures are described. METHODS: Current literature on laparoscopic liver resection is reviewed based on reports identified following a specified PubMed search. RESULTS: Available evidence indicates that laparoscopic liver resection can be made safely in selected patients with comparable duration of surgery, blood loss, tumour clearance, and mortality to that of open resection. Tumours localised peripherally in the left lateral segments of the liver or in segments IV-VI seem to be best suited for laparoscopic resection. The laparoscopic approach may be beneficial to the patients as compared to conventional resection but randomised trials are pending. Laparoscopic resection of colorectal liver metastases is described in a small number of patients only. The long-term outcome following such resections is not adequately documented. CONCLUSIONS: Laparoscopic liver resection is a promising technique with a comparable short-term outcome to that of open procedures but with the potential advantages of minimal invasive treatment. The technique should be further evaluated in properly designed trials. Laparoscopic resection of colorectal liver metastases should not be performed on a regular basis until long-term results are defined. PMID- 16352895 TI - p16 Gene methylation in colorectal tumors: correlation with clinicopathological features and prognostic value. AB - BACKGROUND: Methylation of the CpG islands in the p16 gene promoter region is an important transcription repression mechanism that has been identified as an alternative mechanism for inactivating specific genes in cancer. Given that, DNA methylation is a common event in colorectal cancer. MATERIALS AND METHODS: The aim of this study was to establish the methylation status of the p16 gene in 104 patients with colorectal carcinoma and evaluate its prognostic value. DNA was bisulfite-modified and analyzed for p16 promoter methylation by methylation specific PCR. RESULTS: Methylation of thep16 gene was determined in 18.3% of our patient population (19/104). The methylated state did not correlate with any clinicopathological factors. During a median follow-up period of 72 months, the overall survival rate for patients with gene methylation was 75% and without gene methylation it was 61% (p = 0.09). CONCLUSIONS: Although not statistically significant, this difference indicates a clinically valuable tendency. PMID- 16352896 TI - Reliability study on the Japanese version of the Clinician's Interview-Based Impression of Change. AB - The Japanese version of the Clinician's Interview-Based Impression of Change (CIBIC-plus J) is a semistructured interview format including a 7-point Likert type scale for the Clinician's Global Impression of Change. It comprises subscales of DAD for ADL, Behave-AD for behavioral and psychiatric symptoms and MENFIS for cognitive and emotional impairment. DAD and Behave-AD are used for caregivers and MENFIS for both caregivers and patients. The objective of this study was to examine the interrater reliability of the CIBIC-plus J by the videotape method. Twenty videotaped patients including 13 real and 7 virtual patients were independently assessed by 11 raters. The kappa coefficient from the full set of data of 20 patients and 11 raters was 0.453, and from the limited data of 13 real patients, 0.383. By permitting one-point disagreements on the 7 point scale or by collapsing it into the 3-point scale (improved, no change, or worsened), kappa improved to 0.894 and 0.731, or 0.734 and 0.504, respectively. From these results, it was concluded that the reliability of a 7-point Clinician's Global Impression of Change was halfway, i.e., neither sufficient nor insufficient. However, if one-point disagreements or collapsing into 3-point scale are allowable, the CIBIC-plus J will be sufficiently reliable. PMID- 16352897 TI - The third generation of therapeutic communities: the early development of the TC for addictions in Europe. AB - AIMS: It is the goal of this study to investigate the first development of the drug-free therapeutic community (TC) in Europe. The paper aims at systemizing information, scattered all over Europe and is the first stage in an ongoing study to record the development of the European TC movement and its influences. DESIGN: After a study of the grey (hidden) literature, TC pioneers and experts per country were contacted to further elaborate the first findings. Subsequently, a preliminary summary of our findings was published in the Newsletter of the European Federation of Therapeutic Communities (EFTC), inviting additional information and corrections. The authors completed the results for this article with relevant first-hand information, obtained through interviews with European pioneers. FINDINGS: The findings are summarized under three topics: chronology, interconnections and European identity. It was found that from 1968 until 1989, a new therapeutic approach arose all over Europe, modeled after Synanon, Daytop and Phoenix House, New York, through Phoenix House, London and Emiliehoeve in the Netherlands. Therapeutic communities were established in Belgium, Finland, Germany, Greece, Ireland, Italy, Norway, Spain, Sweden and Switzerland as well. These communities were closely-knit and interconnected in their reaction against psychiatric and methadone treatment. The European TCs developed an own identity compared to the American ones. CONCLUSIONS: The European TCs adapted the model of their American predecessors to their own culture, influenced more by milieu therapy and social learning. Instead of harsh behaviorism, more emphasis was placed on dialogue and understanding. Professionals occupied a more pivotal role and took over the dominant position of ex-addicts. Research, executed by TC professionals gradually entered the TC. A generic network of TC connections, through which the development evolved, was uncovered, and clear regional trends can be observed. PMID- 16352898 TI - Co-morbidity of infectious and addictive diseases in St. Petersburg and the Leningrad Region, Russia. AB - The Russian health care system is organized around specific diseases, with relatively little focus on integration across specialties to address co morbidities. This organizational structure presents new challenges in the context of the recent epidemics of injection drug use (IDU) and HIV. This paper uses existing and new data to examine the prevalence of reported new cases of drug dependence (heroin) and HIV over time as well as associations between drug dependence and alcoholism, hepatitis B and C, and tuberculosis in the City of St. Petersburg and the Leningrad region. We found a sharp rise in reported cases of IDU beginning in 1991 and continuing until 2002/2003, followed by a sharp rise in newly reported cases of HIV. These rises were followed by a drop in new cases of HIV and drug addiction in 2002/2003 and a drop in the proportion of HIV-positive individuals with IDU as a risk factor. Infection with hepatitis B and C were common, especially among injection drug users (38 and 85%, respectively), but also in alcoholics (7 and 14%). Tuberculosis was more common in alcoholics (53%) than in persons with alcoholism and drug dependence (10%), or with drug dependence alone (4%). Though these data have many limitations, they clearly demonstrate that drug dependence and/or alcoholism, HIV, hepatitis, and tuberculosis frequently co-occur in St. Petersburg and the Leningrad Region. Prevention and treatment services across medical specialties should be integrated to address the wide range of issues that are associated with these co morbidities. PMID- 16352899 TI - Survey of smoking attitudes in grade school children in Greece: a preliminary study. AB - AIMS: Tobacco use among children and adolescents is a major public health concern. Past literature has indicated that smoking attitudes are reliable predictors of teen smoking. The present study explored the smoking attitudes of 4th-6th grade Greek pupils. METHODS: Data were obtained from 379 Greek elementary school students who completed a smoking attitudes survey, based on the Bogalusa Heart Study. FINDINGS AND CONCLUSIONS: Analysis of variance indicated that 4th graders expressed significantly more positive attitudes towards tobacco use in three statements related to the psychological consequences of cigarette use, and to peer pressures to smoke. No gender differences were evident. Overall, pupils across grades expressed relatively negative attitudes towards smoking. Differences between school grades in certain statements denoting the psychological properties of tobacco use, and in peer pressures were attributed to 4th graders' lack of knowledge in the particular areas. PMID- 16352900 TI - Parental anti-smoking socialization. associations between parental anti-smoking socialization practices and early adolescent smoking initiation. AB - This longitudinal study of 600 families concentrates on the influence of parental anti-smoking socialization by examining both (a) the effects of eight indicators of anti-smoking socialization on adolescent smoking and (b) the influence of parental smoking on the effectiveness of their anti-smoking socialization. Robust differences between smoking and non-smoking parents demonstrated that both kinds of families hold different norms and attitudes about adolescent smoking and how to deal with it. In terms of effective anti-smoking socialization, it appeared that parental involvement on a more abstract level, such as feeling confident one has influence on the smoking behaviour of one's child and having knowledge whether one's child and his or her friends smoke, seemed important in preventing early adolescent smoking, while concrete communication or house rules about smoking were not. PMID- 16352901 TI - Drugs and social exclusion in ten European cities. AB - AIM: To describe social characteristics seen among socially excluded drug users in 10 cities from 9 European countries, and identify which social exclusion indicators (i.e. housing, employment, education) are most closely linked to intravenous drug use. DESIGN: Cross-sectional survey. SETTING: Interviews were held in social services centers, town halls, streets, squares and other usual meeting points of the target population. PARTICIPANTS: The sample comprises 1,879 participants who have used heroin and/or cocaine and certain derivatives (92.3%) over the last year. Males accounted for 69.7% of the sample, and the mean age was 30.19 years. Participants were recruited in 10 cities: Seville and Granada, Spain; Cologne, Germany; Vienna, Austria; Brussels, Belgium; Athens, Greece; Dublin, Ireland; London, England; Lisbon, Portugal, and Perugia, Italy. MEASUREMENTS: Structured face-to-face questionnaire, conducted by privileged access interviewers. RESULTS: Cannabis, heroin and cocaine are the most widely used substances. In the total sample, 60.2% injected drugs during the last year, 45.9% reported having hepatitis C; 54.9% have been in prison; 14.2% are homeless; 11.3% have a regular job, and 35.2% are involved in illegal activities. Hierarchical logistic regression analysis (injectors and non-injectors) showed that older participants have a greater likelihood of injecting than younger ones. Social exclusion variables associated with intravenous drug use are incarceration, homelessness, irregular employment, and delinquency. Participants who abandoned or were expelled from a drug treatment program are at greater risk of injecting drugs than participants who have never had treatment, are currently in treatment or have been released. CONCLUSION: Personal, social, and economic conditions are all linked in a process of social exclusion that compounds problem drug misuse. Given the findings of this study, we believe that there is a clear need for specific programs targeting specific groups, i.e., distinct strategies must be set in place, in line with the profile and needs of the patient in each context. PMID- 16352902 TI - Alcohol and injury: a risk function analysis from the Emergency Room Collaborative Alcohol Analysis Project (ERCAAP). AB - While emergency room (ER) studies have documented a strong association of alcohol and injury, less is known about the level of risk at which various quantities of alcohol or particular patterns of drinking place the individual for injury. Comparative risk function analyses are carried out in ER samples in seven countries that cover sites in 14 studies included in the Emergency Room Collaborative Alcohol Analysis Project (ERCAAP). Risk of injury is analyzed for the mean number of drinks consumed per day and the number of occasions in which 5 or more drinks were consumed at one time (5+) during the last year. All countries showed similar increases in injury risk to an average volume of about 2 drinks per day, with a leveling off of risk at higher average daily volumes, with the exception of Italy. Risk of injury increased to 12 or more 5+ days for the USA, Canada and Mexico, but leveled off after only 3 5+ days for Argentina and Spain. Poland showed increased risk to 30 5+ days. Similar risk curves were found for both males and females, although females were at lower risk of injury in all countries expect Spain and Poland. In low detrimental drinking pattern societies, risk curves showed higher risk for any drinking and any frequency of 5+ but at higher levels of each, risk levels decreased nearly to levels found for abstainers. Risk functions were also consistent across gender and age groups in low detrimental drinking pattern societies, with higher risk for males and those <30. For those countries with high detrimental drinking patterns, injury risk increased with volume and 5+ drinking primarily among males. This ER-based risk function analysis suggests that risk of injury increases proportionally with increased alcohol consumption at lower consumption levels, but a threshold effect is achieved at relatively low levels of mean daily consumption and higher consumption times. Risk may be culturally specific, dependent, in part, on the manner in which alcohol is used in the culture. PMID- 16352903 TI - Pathophysiology of brain ischemia: penumbra, gene expression, and future therapeutic options. PMID- 16352904 TI - Magnetic resonance investigation of the upper spinal cord in pure and complicated hereditary spastic paraparesis. AB - Neuropathological studies of hereditary spastic paraparesis (HSP) have described axonal loss involving corticospinal and somatosensory tracts in the spinal cord. This MRI-based study was intended to investigate in vivo diameter alterations of the spinal cord in HSP, including both pure HSP (p-HSP, n = 20) and complicated HSP (c-HSP, n = 10). Standard MRI examinations of the cervical and thoracic spinal cord in HSP patients and a control group (n = 54) were analyzed by standardized spinal cord planimetry. In HSP patients, significant atrophy of the upper spinal cord compared to controls was observed at p < 0.001 both at the cervical and at the thoracic level. Myelon diameters at both levels were also significantly reduced in the two HSP subgroups in an additional comparison with age-matched subgroups of controls each, but p-HSP and c-HSP groups themselves did not differ. Marked atrophy of the upper spinal cord seems to be associated with HSP, assumedly due to the central-distal axonopathy. However, the differences between p-HSP and c-HSP could not be visualized by structural MRI at spinal cord level. PMID- 16352905 TI - Upbeat nystagmus due to a small pontine lesion: evidence for the existence of a crossing ventral tegmental tract. AB - We report a patient with an isolated large upbeat nystagmus (UBN) in the primary position of gaze. Eye movements were filmed and recorded using electro oculography. The upward vestibulo-ocular reflex gain, evaluated by pitching the head forward, was markedly reduced compared to when pitching the head back. The lesion was a probable lacunar infarction located in the paramedian and posterior part of the basis pontis, at the upper pons level. This UBN case, with one of the smallest brainstem lesions reported so far, supports the existence in humans of the crossing ventral tegmental tract, described in the cat and transmitting excitatory upward vestibular signals to the third nerve nucleus. It is also suggested that the decussation of this tract lies at the same upper pons level as in the cat but in a slightly more ventral location, i.e. in the posterior basis pontis. PMID- 16352906 TI - Haplotype association analysis of AGT variants with hypertension-related traits: the HyperGEN study. AB - OBJECTIVE: Function of the renin-angiotensin system is important to human hypertension, but its genetic etiology remains elusive. We set out to examine a hypothesis that multiple genetic variants in the system act together in blood pressure regulation, via intermediate phenotypes such as blood pressure reactivity. METHODS: A sample of 531 hypertensive cases and 417 controls was selected from the HyperGEN study. Hypertension-related traits including blood pressure responses to challenges to math test, handgrip and postural change (mathBP, gripBP, and postBP), and body mass index (BMI) were analyzed for association with 10 single nucleotide polymorphisms (SNPs) in the angiotensinogen (AGT) gene. Single-marker and haplotype analyses were performed to examine the effects of both individual and multiple variants. Multiple-trait profiling was used to assess interaction of latent intermediate factors with susceptible haplotypes. RESULTS: In Blacks, two SNPs in exon 5 and 3'UTR showed significant association with gripBP, and two promoter SNPs were strongly associated with postBP. In Whites, only borderline association was found for 2 promoter SNPs with mathBP. Haplotype analyses in Blacks confirmed association with gripBP, and detected significant association of a haplotype to BMI (p=0.029). With the interactions modeled, haplotype associations found in Blacks remain significant, while significant associations to BMI (p=0.009) and gripSBP emerged in Whites. CONCLUSION: Genetic variants in regulatory regions of AGT showed strong association with blood pressure reactivity. Interaction of promoter and genic SNPs in AGT revealed collective action of multiple variants on blood pressure reactivity and BMI both in Blacks and in Whites, possibly following different pathways. PMID- 16352907 TI - On the formula for admixture linkage disequilibrium. AB - Admixture linkage disequilibrium (ALD), a phenomenon created by gene flow between genetically distinct populations, has for some time been used as a tool in gene mapping. It is therefore important to analyze the pattern of ALD over generations. In this study we explore two models of admixture: the gradual admixture (GA) model, in which admixture occurs at a variable rate in every generation; and the immediate admixture (IA) model, a special case of the GA model, in which admixture occurs in a single generation. In the case of ALD, the well-known formula of linkage disequilibrium (Delta(t)=(1-r)t Delta(0)) is not applicable under these two models. We note the effect of a random mating population (RMP) to the gametic frequencies from the parental population to the offspring population, and provide the correct formula for ALD. PMID- 16352908 TI - Odor identification and decline in different cognitive domains in old age. AB - The authors examined the association of odor identification with rate of decline in different cognitive systems. Participants are 481 older persons from the Rush Memory and Aging Project. At baseline, the Brief Smell Identification Test was administered. At annual intervals for up to 3 years, a battery of 19 cognitive tests was administered from which previously established composite measures of 5 cognitive domains were derived. In mixed-effects models adjusted for age, sex, and education, lower odor identification score was associated with lower function at baseline in each cognitive domain. Lower score was also associated with more rapid decline in perceptual speed (estimate=0.015, SE=0.006, p=0.013) and episodic memory (estimate=0.012, SE=0.006, p=0.030) but not with rate of decline in semantic memory, working memory, or visuospatial ability. Thus, on average, a person with a low odor identification score (6, 10th percentile) declined more than twice as rapidly in perceptual speed and episodic memory as a person with a high score (11, 90th percentile). Results were unchanged in subsequent analyses that controlled for cigarette smoking or clinically diagnosed stroke. The results indicate that impaired odor identification in old age is associated with impaired global cognition and more rapid decline in perceptual processing speed and episodic memory. PMID- 16352909 TI - Hallucinations, cognitive decline, and death in Alzheimer's disease. AB - The relation of psychotic symptoms to cognitive decline and mortality in Alzheimer's disease (AD) was examined during a mean of 2.2 years in 478 persons selected from clinical settings. Psychotic symptoms were ascertained at baseline and cognition was assessed semiannually with nine tests from which a global measure was formed. In analyses that controlled for age, sex, race, and education, hallucinations (29.6%), especially visual ones, were associated with more rapid global cognitive decline and increased mortality, even after controlling for baseline level of cognition and use of antipsychotic medication, and the association with mortality increased with higher level of education. Delusions and misperceptions were not strongly related to cognitive decline or mortality. The results suggest that hallucinations in Alzheimer's disease, particularly visual ones, are associated with more rapid progression. PMID- 16352910 TI - Incidence of dementia in very elderly individuals: a clinical, neuropathological and molecular genetic study. AB - AIMS: To evaluate the effect of medical record use on figures for the incidence of dementia and the effect of apolipoprotein E (APOE) polymorphism on this incidence and neuropathologically defined Alzheimer's disease (AD) in very elderly individuals. METHODS: Cognitive functions were examined in a cohort of 328 (92% of the very elderly people of a town participated in this study) nondemented Finnish elderly individuals 85 years of age or more in 1991. The examination was repeated in survivors in 1994, 1996, 1999 and 2001. Medical notes and social work records were evaluated. All these individuals were genotyped for APOE. Neuropathological analysis of AD-type pathology was performed on 159 of 303 subjects who died during the follow-up. RESULTS: Age group, gender or APOE did not significantly affect the incidence of dementia, which was over 20% higher (85 vs. 69 per 1,000 person-years) if the cognitive status at death was ascertained by medical and social work records than without this evaluation. The APOE upsilon4 allele was highly significantly (p=0.002) and age almost significantly (p=0.06) associated with neuropathological AD in nondemented individuals. CONCLUSIONS: Medical records should be analyzed in studies on the incidence of dementia in very elderly individuals. APOE polymorphism does not affect the incidence of dementia in this age group. However, clinical dementia diagnosis in very elderly individuals does not necessarily correlate well with the presence of neuropathological AD which, even in this age group, is significantly associated with the APOE upsilon4 allele. PMID- 16352911 TI - Relationship between lifetime smoking, smoking status at older age and human cognitive function. AB - Cigarette smoking is a major risk factor for clinical cardiovascular disease and may also be associated with poorer cognitive functioning in older age. We measured lifetime cigarette smoking, smoking status and cognitive function in over 2,000 men and women from the general population aged over 50 years with subclinical atherosclerosis (ankle brachial pressure index 0.05); in contrast, NMDA (2.5, 5 and 10 ng intrathecally) can significantly and dose dependently decrease the pain threshold on the hot-plate test (p < 0.05 or p < 0.01) and increase the writhing times (p < 0.05 or p < 0.01) in the mice treated with emulsified anesthetics. These results suggest that spinal NMDA receptors may be important targets for the analgesic effects of emulsified enflurane, isoflurane and sevoflurane. PMID- 16352922 TI - [Recent progress of chemotherapy for colorectal cancer]. AB - The dawn of chemotherapy for gastrointestinal cancers including colorectal cancer was the inception of 5-fluorouraci (l 5-FU), produced by Dr. Heidelberger in 1957. 5-FU had been playing the leading role in chemotherapy for colorectal cancer for four decades until the Nineties. The second wave of chemotherapy was biochemical modulation. For the enhancement of 5-FU effects, the sequential combination with methotrexate and leucovorin (LV) was devised. Among them, 5 FU/LV was established in standard chemotherapy for colorectal cancer, due to several meta-analyses. Oral 5-FU such as UFT, TS-1 and capecitabine derivatives are gaining an important position in chemotherapy for colorectal cancer based on the accumulation of evidence. The third wave enveloped chemotherapy. Irinotecan was developed for gastrointestinal cancer. In 2000, CPT-11+5-FU/LV became the standard and first-line chemotherapy for colorectal cancer in the U.S. and Europe. Unfortunately, we in Japan have lagged behind the U.S. and Europe, because, 5-FU with LV was not approved here until 1999. Oxaliplatin was accepted in Europe first. FOLFOX, including continuous infusion of 5-FU, and LV were approved in 1998. In 2004, oxaliplatin + 5-FU/LV, named FOLFOX 4, was approved as the first-line chemotherapy treatment for colorectal cancer in the U.S. Oxaliplatin was the synthesized in Japan. At last, it was approved with usage restrictions the past March. We are now able to use FOLFOX 4, and this chemotherapy strategy for colorectal cancer in Japan is spreading rapidly. Recently, several new molecular targeted agents, which we call the fourth wave of chemotherapy, have been available in clinical studies,and promising data have been presented. Among them, evacizumab and cetuximab were tested in large phase III studies, their efficacies were upheld, and the drugs were approved in the U.S. Median survival time(MST) has been gradually prolonged through 5-FU/LV with irinotecan and oxaliplatin. Now, with the addition of molecular targeted agents, over 20 months of MST was reported. PMID- 16352923 TI - [Intra-arterial chemotherapy for head and neck cancer]. AB - There are historically speaking, three methods of intra-arterial infusion for head and neck cancer. Recently, daily concurrent chemoradiotherapy using new superselective intra-arterial infusion via superficial temporal arterial artery is noted. A catheter with a curved tip is inserted superselectively to the feeding artery of the tumor via the superficial temporal artery. Long-term catheterization is possible in this method. Thirty-five patients with stage III or IV oral cancer were treated. Radiotherapy (total dose:40 Gy/4 weeks) and superselective intra-arterial infusion chemotherapy using DOC (total dose: 60 mg/m2, 15 mg/m2/week) and CDDP total dose: 100 mg/m2, 5 mg/m2/day) were concurrently performed daily, followed by surgery. In 31 patients, intra-arterial infusion was successful(successful rate: 88.6%), and no major complication was observed. The clinical effects were CR in 25 patients(80.6%), and pathological effects of resected tumor after surgery were pathological CR in 28 (90.3%). This method promises to be new strategy of choice for the treatment of head and neck cancer. PMID- 16352924 TI - [Attempts to improve organ preservation in patients with squamous cell carcinoma of the head and neck]. AB - When surgical resections are performed for patients with locally advanced head and neck cancer, a critical consideration is which organs and functions of patients are sacrificed. In attempts to improve the organ preservation rate in patients with advanced squamous cell carcinoma of the head and neck (SCCHN), chemotherapy has been used either before(neoadjuvant or induction), with (concurrent or concomitant), after (adjuvant) radiotherapy, or as alternating treatment with radiotherapy. A recent systematic review using meta-analysis has revealed that concurrent chemotherapy with radiotherapy shows a significant benefit for the survival rate of patients with SCCHN when compared with radiotherapy alone, and is superior to neoadjuvant chemotherapy. However, no standard concurrent chemoradiotherapy regimen has been defined,although concurrent chemotherapy together with regimens including cisplatin has been considered to be most effective for SCCHN. We combined radiotherapy concurrently with chemotherapy including cisplatin, 5-fluorouracil, methotrexate, and leucovorin in patients with advanced resectable SCC of the hypopharynx to minimize the necessity of radical surgery and preserve the functional larynx. In conclusion, previous reports and our data show that an organ preservation treatment approach using concurrent chemoradiotherapy is feasible. PMID- 16352925 TI - [The role of taxanes for head and neck cancer]. AB - Taxanes are a highly active anti-cancer drug for squamous cell carcinoma of the head and neck (SCCHN). When used as a single agent, the overall response rates were between 20-40%. In Japan, the phase I study revealed that the maximum tolerated dose of the docetaxel was 70-90 mg/m2 and the recommended dose was 60 mg/m2. The main toxicity was neutropenia. Docetaxel administered in combination with cisplatin has demonstrated promising response rates ranging from 40-70% for recurrent or metastatic SCCHN. Its effect is similar to that of a cisplatin plus 5-fluorouracil regimen. A three-drug combination with docetaxel, cisplatin and 5 fluorouracil has been tested. The preliminary results with three drugs in combination seems superior to a two-drug combination. However, the final conclusions regarding theses drug combinations will be given by the results of an ongoing EORTC #249 71 study. Weekly docetaxel with concurrent chemoradiotherapy is feasible and active for locally unresectable SCCHN. Cisplatin plus 5 fluorouracil based concurrent chemoradiation regimens have demonstrated improved survival compared with conventional radiotherapy. Docetaxel including regimens showed efficacy with mucositis as the major acute toxicity and a swallowing problem as the chronic toxicity. Thus, the three-drug combination as the chemoradiation setting seems to be toxic for Japanese patients. A new intensive supporting care approach focused on mucositis is warranted. PMID- 16352926 TI - [Combination therapy with TS-1]. AB - The combination with cisplatin (CDDP) and 5-FU is considered the chemotherapy of choice for squamous cell carcinoma of the head and neck (HNSCC). TS-1, a modulation of tegafur developed in Japan, is an orally administered active agent for HNSCC. Some clinical phase I/II studies on the combination of CDDP and TS-1 have been reported. The combination showed a good response rate, 67.6% for both advanced and recurrent HNSCC, in our clinical phase II study. Regimens of TS-1 combined with carboplatin or nedaplatin are also reported.TS-1 containing regimens appear to be effective for HNSCC, and multi-institutional phase II studies with large sample size are needed in future. The combination TS-1 and radiotherapy, its dose and schedule,are being studied in phase I trials for advanced HNSCC. Patient compliance is better than with 5-FU injection because TS 1 is orally administered. The adverse effect, especially in terms of bone marrow toxicity, is equal or better than with 5-FU injection. The TS-1 combination with radiotherapy is a useful regimen for outpatients. The efficacy and adverse effects should be studied in carefully designed phase I/II trials. TS-1 will be one of the key drugs for HNSCC in future. PMID- 16352927 TI - [Gene therapy and molecular therapeutic for head and neck cancer]. AB - Despite recent progress in radiotherapy, chemotherapy and surgical techniques, the survival of patients with head and neck squamous cell carcinomas has not improved significantly over the past three decades. To conquer these malignant tumors, various new therapies have been under development, including gene therapies. In this paper, we have reviewed recent advances in gene therapies and molecular target therapy for head and neck cancers, with a special focus on the replication-selective adenovirus vector. At present, replication-defective adenovirus-based vectors have been widely used as convenient and safe vectors for transferring therapeutic genes into target cancer cells. However, these vectors have limited the efficacy of treatment by restricting the number of tumor cells to which the therapeutic gene can be delivered. Replication-selective adenoviral vectors (RSAV) are adenoviruses designed to have a limited ability to replicate themselves in the targeted tumor cells but not in other normal tissues. Tumor cell killing is achieved not by the genes delivered by the vectors but by the oncolysis induced by the replicated viruses by their original nature as adenovirus. Amplified viral vectors also spread to the adjacent tumor cells and kill these cells in the same manner. Recently, increased evidences of antitumor activity of RSAVs have been demonstrated preclinically, and several clinical trials have demonstrated the safety and clinical activity of replication selective viruses. PMID- 16352928 TI - [Pharmacokinetic comparison of capecitabine and 5'-deoxy-5-fluorouridine (doxifluridine; 5'-DFUR)]. AB - Capecitabine is a new oral fluoropyrimidine derivative. It is a prodrug that is activated to fluorouracil in three metabolic steps. Compared to 5'-deoxy-5 fluorouridine (doxifluridine) itself, capecitabine was designed to deliver more doxifluridine to cancer cells,where it is activated to fluorouracil. Based on differential distribution of the three metabolizing enzymes in different tissues, capecitabine is considered to yield more fluorouracil in cancer cells than bone marrow cells or gastro-intestinal epithelial cells. Due to these pharmacokinetic merits, capecitabine can be given at a higher dose, and therefore is expected to yield higher antitumor activity, than doxifluridine. To investigate the pharmacokinetic advantage of capecitabine compared to doxifluridine, pharmacological studies of both drugs have been conducted in phase II trials. These used the same study design,and compared the pharmacokinetics and pharmacodynamics between capecitabine and doxifluridine. The area under the curve of doxifluridine was 2.3 times higher after the administration of capecitabine than doxifluridine, suggesting that higher intratumor concentrations of fluorouracil were indeed achieved after capecitabine administration. This study clearly documents the pharmacokinetic superiority of capecitabine compared to doxifluridine. PMID- 16352929 TI - [Autologous hematopoietic stem cell transplantation for aggressive B-cell non Hodgkin's lymphoma]. AB - To evaluate the results of high-dose chemotherapy (HDT) and autologous hematopoietic stem cell transplantation (ASCT) in patients with diffuse B-cell aggressive non-Hodgkin's lymphoma(NHL). Between 1991 and 2004, 25 patients who did not achieve complete remission and 26 in complete remission from conventional chemotherapy received HDC-ASCT. Of 25 patients with refractory NHL,14 were chemotherapy-sensitive before HDT-ASCT and 11 were chemotherapy-resistant. CR was achieved after HDC-ASCT in 50% of 14 chemotherapy sensitive patients and in none of 11 chemotherapy-resistant patients. The 5-year probability of event-free survival for chemotherapy-sensitive and chemotherapy-resistant patients was 51.3% and 20.8%, respectively (p<0.05, log-rank test). Moreover, the 5-year probability of event-free survival for patients in the low-risk group with International Prognostic Index (IPI) and in the high-risk group with IPI was 75.0% and 16.3%, respectively (p<0.05, log-rank test). HDT-ASCT should be considered for patients with refractory aggressive NHL who are chemotherapy-sensitive rather than chemotherapy-resistant. Twenty-six patients in complete remission received consolidation therapy with HDT-ASCT. The 5-year probability of disease-free survival for patients in the low-risk group and in the high-risk group was 68.8% and 60.0%,respectively (p = 0.9 6). HDT-ASCT should be considered for patients at high risk who achieve complete remission after induction treatment. In future, HDT-ASCT combined with rituximab as induction therapy or as consolidation therapy is needed for patients with aggressive NHL in the high-risk group. PMID- 16352930 TI - [Clinical phase trial of concurrent chemoradiotherapy combined TS-1 and nedaplatin]. AB - We have treated head and neck carcinoma by concurrent chemoradiotherapy combined with 5-fluorouracil (5-FU) and cisplatin (CDDP). However,this chemoradiotherapy could not show an enormous effect in the advanced carcinoma of Stage III and IV. Therefore,we changed the contents of the chemotherapy, i.e., we replaced 5-FU, one of the agents with time dependency, to continuous administration of TS-1 for 2 weeks,also replacing CDDP, one of the agents with dose dependency, to nedaplatin (CDGP) in order to reduce kidney dysfunction. In this concurrent chemoradiotherapy, oral TS-1 was continued for 2 weeks and CDGP was administered on the 4 th day from the start of TS-1. In addition, radiotherapy was performed concurrently. In this way,we performed a phase I clinical trial of concurrent chemoradiotherapy combining TS-1 and nedaplatin (CDGP). As for the incidence of adverse events,grade 3 mucositis due to radiation was observed in two patients. As a result of the phase I clinical trial,we decided the maximum-tolerated dose (MTD) of TS-1 to be 80 mg/m2 (maximum 120 mg/body) and 100 mg/m2 for CDGP, and then determined the recommended dose(RD) of TS-1 as 80 mg/m2 (maximum 120 mg/ body) TS-1 and of CDGP as 9 0 mg/m2 CDGP. PMID- 16352931 TI - [Assessment of goserelin treatment in adjuvant therapy for premenopausal patients with breast cancer in Japan-zoladex breast cancer study group trial-B]. AB - OBJECTIVE: Goserelin (GOS) therapy in an adjuvant setting for estrogen receptor(ER)-positive premenopausal patients with breast cancer was assessed in a randomised comparative study. METHODS: ER positive premenopausal patients with n + or n 0 and T > or = 3 cm received tamoxifen (TAM) 20 mg/day, GOS 3.6 mg/4 weeks or GOS + TAM for 2 years, and the clinical efficacy and safety of these regimens were assessed. RESULTS: In the data analysis of total 207 patients, hazard ratios of disease free survival (DFS) and overall survival (OS) in the GOS group compared to the TAM group were 0.87 and 2.10,respectively. The incidence of adverse drug reactions was similar (42-55%) in all three groups. Since the number of patients in this study did not reach the target number, the efficacy could not be assessed from a statistical aspect. Therefore,meta-analysis with similar foreign studies(ZIPP) was implemented. The results of meta-analysis showed that the hazard ratios of DFS and OS in the GOS group compared to the non-GOS group were 0.83 and 0.85, respectively. CONCLUSION: Although the analysis of 207 patients did not show any statistically significant difference between each of the treatment groups, the results of meta-analysis showed a significant prolongation of DFS in the GOS group. Also high tolerability of GOS was suggested. From these results, GOS was considered highly useful in adjuvant therapy for ER-positive premenopausal patients with breast cancer. PMID- 16352932 TI - [Neoadjuvant chemotherapy using TS-1 and CDDP against large type 3/Type 4/Bulky N 2 advanced gastric cancer]. AB - This study was conducted to assess therapeutic results following neoadjuvant chemotherapy (NAC) for large type 3/type 4/Bulky N 2 advanced gastric cancer having a poor prognosis following resection. The subjects consisted of cases (< or = 75 y.o.) having large type 3 (diameter > or = 8 cm), type 4 or Bulky N 2 gastric cancer curable by resection based on preoperative imaging diagnostics. The NAC regimen consisted of TS-1 at 80-120 mg/body on days 1-21 p. o. and CDDP at 60 mg/m2 on day 8 divided. Upon completion of two courses of 4 weeks per course, gastrectomy with > or = D2 lymph node dissection was carried out on days 21-34. The average age of the subjects was 60.7 years, and the therapy completion rate was 80% (8/10 cases). Five of ten cases were responders diagnosed as grade 2 by histopathological examination of excised specimens (response rate 50%). Two of five responders were histopathologically evaluated as down-staging as a result of NAC (Stage III A--> f Stage I A, Stage IV--> f Stage I A). Three of the five non responders have relapsed, and the relapse-free interval was an average 238 days. In the five responders,one has relapsed at 331 days,while the other 4 responders have shown no relapse yet. Although NAC consisting of TS-1 and CDDP is considered to be effective against advanced gastric cancer, a phase III study with surgical treatment only will be necessary to confirm its true value. PMID- 16352933 TI - [Clinical study of ambulatory patient cancer chemotherapy for advanced colorectal cancer]. AB - To evaluate ambulatory patient cancer chemotherapy, the clinical response, toxicities and survival time were analysed among 19 patients with non-curative or recurrent colorectal cancer who were treated by Uracil/Tegafur (UFT) plus oral Leucovorin (UZEL) for the past 2 years. The patients were administered UFT (300 mg/m2/day) and UZEL (75 mg/body/day) for 28 days with a one-week interval every 35 days as one course.A partial response (PR) was observed in 6 patients (31.6%) and stable disease (SD) in 8. The median survival time was 16 months. Although nausea/vomiting, diarrhea and leucopenia were noted, no severe side effects were observed. These results suggested that UFT plus Leucovorin therapy might be a useful cancer chemotherapy for ambulatory patients with advanced colorectal cancer. PMID- 16352934 TI - [A randomized crossover study of ramosetron plus dexamethasone for the prevention of nausea and vomiting induced by chemotherapy including cisplatin-comparison of ramosetron combined with 8 mg and 12 mg of dexamethasone]. AB - A randomized crossover study between 0.3 mg of ramosetron (RAM) combined with 8 mg of dexamethasone (DEX) and 0.3 mg of RAM combined with 12 mg of DEX was performed to investigate the prevention of nausea and vomiting due to chemotherapy including 60 mg/m2 or 70 mg/m2 of cisplatin (CDDP) in patients with advanced head and neck squamous cell carcinoma (HNSCC). Twenty-five patients the study consisted of who received chemotherapy with CDDP were enrolled in the present study between January 2001 and December 2002 at the Yokohama City University School of Medicine or Yokohama City University Medical Center. The antiemetic effectiveness in the group receiving 12 mg of DEX was not significantly superior to the group receiving 8 mg of DEX. It was suggested that the antiemetic therapy of RAM 0.3 mg plus DEX 8 mg was effective for the prevention of nausea and vomiting induced by CDDP in patients with advanced HNSCC. PMID- 16352935 TI - [Improved quality of life in a patient with carcinomatous meningitis from invasive lobular carcinoma treated with intrathecal MTX and Ara-C in combination with systemic docetaxel]. AB - Carcinomatous meningitis from breast cancer is rare and has a poor prognosis with a life expectancy of approximately 80 days. We performed intrathecal MTX and Ara C in combination with systemic docetaxel treatments in the patient, a 44-year-old woman with carcinomatous meningitis from invasive lobular carcinoma previously treated with anthracycline and paclitaxel. Although the patient died 128 days after the onset, intrathecal chemotherapy in combination with systemic docetaxel reduced the number of malignant cells and induced degeneration of the cells in CSF, resulting in improved quality of life. PMID- 16352936 TI - [A case of curatively resected schirrous gastric cancer effectively treated by combined chemotherapy of weekly paclitaxel (PTX) and CDDP]. AB - The patient was a 40-year-old woman who had unresectable advanced gastric cancer with invasion of liver and dissemination to the peritoneal wall. Neoadjuvant chemotherapy consisting of paclitaxel (PTX) and CDDP was performed. PTX (80 mg/m2), and CDDP (25 mg/m2) was administered weekly on day 1, 8 and 15 as one cycle. After four cycles of PTX/CDDP administration,invasion of liver and gastric tumor had decreased to almost normal size, and total gastrectomy was performed. After operation, the patient was treated with six courses of PTX/CDDP and survived without recurrence as of this writing. PTX/CDDP was associated with few adverse events in hospital visits, and thought to be an effective chemotherapy against advanced gastric cancer. PMID- 16352937 TI - [Radiation therapy leading to complete response of residual lymph node metastasis in advanced gastric cancer]. AB - A 74-year-old male was admitted to Sendai Saiseikai Hospital with a complaint of upper abdominal pain. Gastrointestinal fibroscopy showed type 2 gastric cancer in the lower third of the stomach,and distal gastrectomy and D 2 lymph node dissection were performed. Metastatic lymph node 8 a had severely infiltrated the pancreas, and pancreatoduodenectomy was required to complete the curative surgery. However, because of preoperative complications of interstitial pneumonia, metastatic node 8 a remained untreated. Postoperatively, radiation therapy (38 Gy plus a boost of 18 Gy) for residual lymph node metastasis was performed over a one-month period. The metastatic node underwent complete regression following radiation therapy, and the patient was discharged without any adverse effects. He received TS-1 medication as postoperative adjuvant therapy. Four years have passed since the complete regression of the lymph node,and the patient has shown no signs of relapse. Therefore, our case suggests that postoperative radiation therapy can be a useful tool for treatment of residual lymph node metastasis in gastric cancer. PMID- 16352938 TI - [Effective TS-1+paclitaxel administration for gastric cancer with malignant ascites--a case report]. AB - We report a case in which TS-1 + paclitaxel (PTX) administration was effective for gastric cancer with malignant ascites. The patient was a 66-year-old male who received total gastrectomy, distal pancreatectomy and splenectomy. He complained of abdominal fullness and ascites 18 months later. The administered regimen of chemotherapy was TS-1 100 mg/day for two weeks, and PTX 120 mg/day on day 1 and 8 of TS-1 intake, followed by 1-week rest. Computed tomography (CT) showed complete loss of malignant ascites. The toxic events were grade 2 leukopenia and grade 2 alopecia. PMID- 16352939 TI - [A case of gastric cancer with multiple liver metastases effectively treated by hepatic arterial infusion of MMC, TS-1/CDDP therapy, and thermotherapy]. AB - A 62-year-old man was revealed to have type 2 gastric cancer with synchronous liver metastasis. We considered liver metastasis to be a prognostic factor, and performed two courses of combination thermochemotherapy consisting of hepatic arterial infusion of MMC and TS-1 and thermotherapy. Partial response was observed in the liver metastases,but the primary lesion showed no changes; therefore, we performed four courses of combination thermochemotherapy consisting of TS-1/CDDP therapy and thermotherapy. By the end of three courses of this therapy,the primary lesion had cicatrized,and endoscopic biopsy revealed no cancer cells. These results suggest that gastric cancer,in which liver metastasis is considered to be a prognostic factor,can be effectively treated by combination therapy with hepatic arterial infusion therapy, followed by thermochemotherapy for the primary lesion. PMID- 16352940 TI - [A case of stage IV gastric cancer previously treated with TS-1 completely responding to second-line chemotherapy with weekly paclitaxel therapy]. AB - We report herein a case with stage IV gastric cancer previously treated with TS-1 completely responding to second-line chemotherapy with weekly paclitaxel therapy. A 65-year-old female was diagnosed as having type 3 gastric cancer with para aortic lymph node metastases. She underwent total gastrectomy with extended lymph node dissection on March 2003. Histopathological examination revealed that the tumor was poorly-differentiated adenocarcinoma with para-aortic lymph nodes metastases and completely resected. After the operation,she was treated by adjuvant chemotherapy with TS-1. In March of 2004, she suffered from hematuria, and a CT scan revealed para-aortic lymph nodes metastases and left kidney metastasis. Then, she was treated by a weekly infusion of paclitaxel as second line chemotherapy. After 3 courses, the tumor disappeared and efficacy was judged as CR. Moreover, CR was maintained after 7 courses. At this writing in January of 2005, she is well and has been treated with paclitaxel without any severe adverse events. Therefore, weekly paclitaxel therapy was considered to be one of the promising second-line chemotherapies for advanced or recurrent gastric cancer previously treated by TS-1. PMID- 16352941 TI - [A case of synchronous gastric and hepatocellular carcinoma successfully treated by TS-1 and hepatic arterial infusion chemotherapy (HAI) of low-dose CDDP]. AB - A 75-year-old man underwent distal gastrectomy for advanced gastric cancer with liver and lymph node metastases and synchronous hepatocellular carcinoma in April 2004. HAI with low-dose CDDP/TS-1 combination therapy was initiated after gastrectomy. Liver and lymph node metastases decreased significantly, with achievement of a partial response (PR) and a complete response (CR), respectively, and the hepatocellular carcinoma was reduced to 54.1% of its initial size after 3 sessions of this chemotherapy. These results suggested that combined chemotherapy with TS-1 and HAI with low-dose CDDP was not only useful for liver and lymph node metastases from gastric cancer, but for hepatocellular carcinoma as well. PMID- 16352942 TI - [Peritoneal dissemination from gastrointestinal stromal tumor of small intestine responding completely to imatinib mesylate (STI 571)]. AB - The prognosis of metastatic or recurrent GISTs is poor, because these tumors resist chemotherapy and radiotherapy. We report a patient with recurrent GIST who underwent molecularly targeted therapy with imatinib, a novel oral tyrosine kinase inhibitor. A 64-year-old man presented with large intra-abdominal mass. The patient had a history of jejunostomy with colostomy for intestinal GIST. The abdominal mass was phi3 x 3.5 cm in size with ascites at Douglas, as determined by computed tomography, and was diagnosed as a peritoneal relapse of GIST. Treatment with imatinib daily was started. After 1 month of treatment with imatinib, reduction of the abdominal tumor began to be recognized on palpation. Computed tomographic scanning at 11 months revealed that the tumor had completely disappeared. The major side effect was drug eruption,which was easily manageable with 2 weeks drug holidays. Imatinib shows promise as a safe and effective drug for the treatment of patients with recurrent GISTs. PMID- 16352943 TI - [A case of recurrent colon cancer responding completely to uracil/tegafur (UFT) plus oral leucovorin (LV) therapy]. AB - The patient was a 78-year-old woman who underwent right hemicolectomy with lymph node dissection (D 2) for cecal cancer in June 2003. Histological diagnosis was moderately differentiated adenocarcinoma, ss, n(-), P 0, H 0, M (-), stage II. Adjuvant chemotherapy was not conducted, and the patient was periodically observed after operation. In April 2004, the serum CEA level was elevated to 14.9 ng/ml. Abdominal CT examinations revealed a tumor, 50 x 35 x 50 mm in size, on the right iliopsoas muscle close to the anastomotic site. Systemic chemotherapy with UFT + Leucovorin was initiated under the diagnosis of local recurrence. Only grade 1 body weight loss,pigmentation, pruritus, and anorexia were recognized during chemotherapy. However, these complications did not require administration. After completion of 6 courses of this chemotherapeutic regimen,the serum CEA level was normalized at 3.1 ng/ml, and CT scan revealed the tumor had disappeared in November 2004. Currently, the patient is free from any signs of recurrence and has maintained a complete remission (CR). PMID- 16352944 TI - [An outpatient with unresectable pancreatic cancer treated with gemcitabine showing prolonged NC (22 months)]. AB - A 76-year-old man developed jaundice and was hospitalized in January 2002. A 3 cm tumor was found in the head of the pancreas by abdominal CT, and the patient underwent laparotomy. The tumor was histologically diagnosed as a well differentiated adenocarcinoma, and showed extensive invasion to the portal vein (T4NXM 0 Stage IV a). Incisional biopsy and hepaticojejunostomy were performed. On the basis of a drug sensitivity test, chemotherapy with 800 mg/m2/week gemcitabine was administered. The patient showed prolonged NC without any symptoms for 22 months, although the CEA and DUPAN-2 levels gradually increased during this time and massive ascites were detected in a routine abdominal CT at 22 months postsurgery. The patient died after 25 months of chemotherapy. Here we report a case of unresectable pancreatic cancer treated with gemcitabine on the basis of a drug sensitivity test. PMID- 16352945 TI - [A paclitaxel-resistant case of recurrent breast cancer responded to combination therapy of capecitabine and trastuzumab]. AB - The patient was a 72-year-old female. Under the supervision of her former doctor, this patient had an operation and adjuvant chemotherapy for progressive breast cancer. During the following period, local recurrence of breast cancer and pulmonary lymphopathia developed. Although medication with paclitaxel was attempted, the focus was resistant to this treatment, and metastasis to the brain was also observed. Due to the dyscrasia above, the patient had difficulty breathing and became bedridden. Subsequently, combination treatment of capecitabine and trastuzumab was attempted. As a result,metastasis in the brain and pulmonary lymphopathia were improved. The patient recovered enough to be discharged at one time. However, his condition took a turn for the worse after the interruption of the combination treatment by a side effect. In conclusion, the combination treatment of capecitabine and trastuzumab is thought to be effective for non-responders to paclitaxel. PMID- 16352946 TI - [Effective salvage chemotherapy with S-1 alone in a patient with lung metastasis of breast cancer]. AB - A patient with lung metastasis of breast cancer was reported. The patient underwent surgery in December, 1999. Her breast cancer then recurred in December, 2000. After treatment failure with anthracycline and taxane antitumor drugs,she participated in a phase II study of S-1, a fluorinated pyrimidine anticancer drug, which was given orally at 80 mg/m2/day (2 doses). After completion of 4 courses of treatment,the target lesions of the lung metastasis markedly shrunk by 47.5% as compared with the pretreatment. Because salvage therapy with S-1 alone showed good antitumor efficacy and beneficial tolerability when the standard dosage was maintained, it was considered that this home therapy was effective for advanced/recurrent breast cancer that was resistant to anthracycline and taxane antitumor drugs. PMID- 16352947 TI - [CPT-11-induced diarrhea treated with indisetron hydrochloride--a case report]. AB - We report a case in which 5-HT3 receptor antagonist, indisetron hydrochloride,was effective against CPT-11-induced diarrhea. When the patient, a 63-year-old male, developed lymph node metastases after resection for primary gastric cancer, 24 hour continuous administration of CPT-11 (125 mg/m2) was initiated. The following day he had diarrhea, possibly associated with the CPT-11. Hange-shashinto was administered but did not improve the condition. Thereafter, the diarrhea followed a protracted course (grade 1 to 2). His diarrhea improved when indisetron hydrochloride was administered with the fifth course of chemotherapy in place of the usual antiemetic drug. In this patient, indisetron hydrochloride, in addition to its role as an antiemetic, was considered to be effective against CPT-11 induced diarrhea. PMID- 16352948 TI - Pulse total-hemoglobinometer provides accurate noninvasive monitoring. AB - OBJECTIVE: Rapid noninvasive measurement of total hemoglobin would be extremely useful for various clinical situations. This study determined the clinical accuracy and utility for a pulse total-hemoglobinometer using four wavelengths: 660 nm (reduced hemoglobin), 805 nm (isosbestic point), 940 nm (oxygenated hemoglobin), and 1300 nm (water density). DESIGN: Clinical trial. SETTING: University school of medicine. PATIENTS: Patients were 122 individuals (age, 18 82 yrs; 49.4 +/- 16.0 yrs [mean +/- SD]), including 71 healthy volunteers, 24 patients undergoing surgery, and 27 patients undergoing hemodialysis. INTERVENTIONS: The hemoglobinometer probe, which simultaneously indicated peripheral oxygen saturation, pulse rate, and hemoglobin, was placed on the fingertip similarly to a regular pulse oximeter. The hemoglobin values were compared with those obtained by the co-oximeter or the sodium lauryl sulfate methemoglobin method. Those hemoglobin values were assigned to either the training set or the validation set for statistical evaluation. MEASUREMENTS AND MAIN RESULTS: Multiple regression analysis including the ratio of the pulsatile optical density (phi(ij)) derived from the four wavelengths and other factors demonstrated that the mean value of the normalized pulse wave obtained from the photodiode at 805 nm (DC805) and the ratios of DC940 and DC1300 (DC940/DC1300) were the pivotal factors in the hemoglobinometer's increased accuracy in the clinically useful range. The coefficient of determination between both methods was r2 = .81 (p < .0001) in the training set and r2 = .75 (p < .0001) in the validation set. When the cutoff value of anemia was set at 10 g/dL, and anemia was defined as <10 g/dL, the respective sensitivity and specificity of hemoglobinometer values to detect anemia in intraoperative patients were 84.3% and 84.6% (n = 20). CONCLUSIONS: The data demonstrated the necessity for consideration of light scattering in red blood cells for pulse-spectrophotometric hemoglobin measurement. This was accomplished with additional factors, such as DC805 and DC940/DC1300. With these improvements, the pulse hemoglobinometer provided noninvasive, clinically acceptable measurement of hemoglobin. The pulse hemoglobinometer is a versatile tool that might be useful for routine health checkups of neonates and young children, intraoperative monitoring of bleeding, and emergency care. PMID- 16352951 TI - Right Care, Right Now--you can make a difference. PMID- 16352952 TI - The intensivist in a spiritual care training program adapted for clinicians. AB - BACKGROUND: Critical illness is a crisis for the total person, not just for the physical body. Patients and their loved ones often reflect on spiritual, religious, and existential questions when seriously ill. Surveys have demonstrated that most patients wish physicians would concern themselves with their patients' spiritual and religious needs, thus indicating that this part of their care has been neglected or avoided. With the well-documented desire of patients to have their caregivers include the patient's spiritual values in their health care, and the well-documented reality that caregivers are often hesitant to do so because of lack of training and comfort in this realm, clinical pastoral education for health care providers fills a significant gap in continuing education for caregivers. OBJECTIVES: To report on the first 6 yrs of a unique training program in clinical pastoral education adapted for clinicians and its effect on the experience of the health care worker in the intensive care unit. We describe the didactic and reflective process whereby skills of relating to the ultimate concerns of patients and families are acquired and refined. DESIGN AND SETTING: Clinical pastoral education designed for clergy was adapted for the health care worker committed to developing skills in the diagnosis and management of spiritual distress. Clinician participants (approximately 10-12) meet weekly for 5 months (400 hrs of supervised clinical pastoral care training). The program is designed to incorporate essential elements of pastoral care training, namely experience, reflection, insight, action, and integration. RESULTS: This accredited program has been in continuous operation training clinicians for the past 6 yrs. Fifty-three clinicians have since graduated from the program. Graduates have incorporated clinical pastoral education training into clinical medical practice, research, and/or further training in clinical pastoral education. Outcomes reported by graduates include the following: Clinical practice became infused with new awareness, sensitivity, and language; graduates learned to relate more meaningfully to patients/families of patients and discover a richer relationship with them; spiritual distress was (newly) recognizable in patients, caregivers, and self. CONCLUSIONS: This unique clinical pastoral education program provides the clinician with knowledge, language, and understanding to explore and support spiritual and religious issues confronting critically ill patients and their families. We propose that incorporating spiritual care of the patient and family into clinical practice is an important step in addressing the goal of caring for the whole person. PMID- 16352953 TI - Hydrocortisone increases the sensitivity to alpha1-adrenoceptor stimulation in humans following hemorrhagic shock. AB - OBJECTIVE: To assess the pressor response to phenylephrine infusion before and after hydrocortisone in severe trauma patients and to correlate this response with their adrenal reserve. DESIGN: Prospective clinical study. SETTING: Surgical intensive care unit in a university teaching hospital. PATIENTS: Twenty-three young trauma patients (Injury Severity Score, 38 +/- 14) were studied at the end of the resuscitative period (27 +/- 15 hrs after trauma). INTERVENTIONS: Total cortisol response to intravenous corticotropin bolus (250 microg) was obtained. Total cortisol response <9 microg/dL defined an impaired adrenal function and the patient was called a nonresponder. Twelve to 24 hrs following this stimulation, phenylephrine was infused in a stepwise manner to establish the phenylephrine mean arterial pressure dose-response curve before and after intravenous hydrocortisone administration (50 mg). An Emax model was used to describe the curve; the influence of the group (responder/nonresponder), the sequence (before/after hydrocortisone), and three covariates (Injury Severity Score, shock, and interleukin-6) were thereafter tested. MEASUREMENTS AND MAIN RESULTS: Forty-three percent of patients were nonresponders. Total cortisol response was not correlated with serum albumin concentration and was negatively correlated with the interleukin-6 concentration. A trend for a higher incidence of nonresponders (53% vs. 36%) and a lesser total cortisol response (7.9 +/- 5.1 vs. 12.5 +/- 5.1 microg/dL) was observed in the shock patients. A phenylephrine dose response structure (E0, ED50, and Emax) was described without influence of the group and the sequence. However, hydrocortisone induced a 37% decrease in ED50 without change in Emax in the shock patients. CONCLUSION: An acute administration of hydrocortisone increases the sensitivity to alpha1-adrenoceptor stimulation in fully resuscitated severe trauma patients following hemorrhagic shock. This effect is independent of the adrenal reserve of the patients and different from that previously reported in septic patients. PMID- 16352954 TI - Induction of hypothermia in patients with various types of neurologic injury with use of large volumes of ice-cold intravenous fluid. AB - OBJECTIVE: Mounting evidence suggests that mild to moderate hypothermia can mitigate neurologic and myocardial injury. The speed of induction appears to be a key factor in determining its efficacy. However, even when the fastest currently available cooling techniques are used, reaching target temperatures takes at least 2 hrs and usually longer. We hypothesized that infusion of refrigerated fluids could be a safe accessory method to increase cooling speed. DESIGN: Prospective intervention study. SETTING: University teaching hospital. PATIENTS: One hundred thirty-four patients with various types of neurologic injury (postanoxic encephalopathy, subarachnoid hemorrhage, or traumatic brain injury). MEASUREMENTS AND MAIN RESULTS: Hypothermia was induced in 134 patients with various types of neurologic injury, by means ice-water cooling blankets and infusion of refrigerated (4 degrees C) saline (110 patients) or saline and colloids (24 patients). An average volume of 2340 +/- 890 mL of refrigerated fluids was infused in 50 mins. Core temperatures decreased from 36.9 +/- 1.9 degrees C to 34.6 +/- 1.5 degrees C at t = 30 mins and to 32.9 +/- 0.9 degrees C at t = 60 mins (target temperature: 32 degrees C-33 degrees C). Monitoring of blood pressure, heart rhythm, central venous pressure, blood gasses, electrolyte and glucose levels, and platelet and white blood cell count revealed no additional adverse effects. Mean arterial pressure increased by 15 mm Hg, with larger increases in blood pressure occurring in hemodynamically unstable patients. No patient developed pulmonary edema. CONCLUSIONS: Induction of hypothermia by means of cold-fluid infusion combined with ice-water cooling blankets is safe, efficacious, and quick. Because the speed of cooling is important to increase its protective effects, we recommend that cold-fluid infusion be used in all patients treated with induced hypothermia. This should be combined with another method to safely and accurately maintain hypothermia once target temperatures have been reached. PMID- 16352955 TI - Evidence of myocardial hibernation in the septic heart. AB - OBJECTIVE: Myocardial hibernation is an adaptive response to ischemia and hypoxia. Hibernating cardiomyocytes are reversibly hypocontractile and demonstrate characteristic metabolic and ultrastructural changes. These include a switch in primary substrate utilization from fatty acids to glucose, up regulation of the myocardial specific glucose transporters (GLUT1 and GLUT4), and glycogen deposition within and between cardiomyocytes. We hypothesized that myocardial hibernation may underlie sepsis-associated myocardial depression. DESIGN: Prospective observational study aimed at identifying the characteristic changes of hibernation in the septic heart. SETTING: University hospital-based laboratory. SUBJECTS: Forty-three C57Bl6 male mice. INTERVENTIONS: Mice underwent cecal ligation and double puncture, sham operation, or no operation and were evaluated 48 hrs after the procedure. MEASUREMENTS AND MAIN RESULTS: Using novel, clinically relevant technology such as magnetic resonance imaging, positron emission tomography, and single photon emission computed tomography imaging, we found septic mice to have diminished cardiac performance, increased myocardial glucose uptake, increased steady-state levels of myocardial GLUT4, and increased deposits of glycogen, recapitulating the changes during hibernation. Importantly, these changes occurred in the setting of preserved arterial oxygen tension and myocardial perfusion. CONCLUSIONS: Sepsis-associated cardiac dysfunction may reflect hibernation. Furthermore, such down-regulation of cellular function may underlie sepsis-induced dysfunction in other organ systems. PMID- 16352956 TI - Fluid responsiveness in spontaneously breathing patients: a review of indexes used in intensive care. AB - OBJECTIVE: In spontaneously breathing patients, indexes predicting hemodynamic response to volume expansion are very much needed. The present review discusses the clinical utility and accuracy of indexes tested as bedside indicators of preload reserve and fluid responsiveness in hypotensive, spontaneously breathing patients. DATA SOURCE: We conducted a literature search of the MEDLINE database and the trial register of the Cochrane Group. STUDY SELECTION: Identification of reports investigating, prospectively, indexes of fluid responsiveness in spontaneously breathing critically ill patients. All the studies defined the response to fluid therapy after measuring cardiac output and stroke volume using the thermodilution technique. We did not score the methodological quality of the included studies before the data analysis. DATA EXTRACTION: A total of eight prospective clinical studies in critically ill patients were included. Only one publication evaluated cardiac output changes induced by fluid replacement in a selected population of spontaneously breathing critically ill patients. DATA SYNTHESIS: Based on this review, we can only conclude that static indexes are valuable tools to confirm that the fluid volume infused reaches the cardiac chambers, and therefore these indexes inform about changes in cardiac preload. However, respiratory variation in right atrial pressure, which represents a dynamic measurement, seems to identify hypotension related to a decrease in preload and to distinguish between responders and nonresponders to a fluid challenge. CONCLUSIONS: Further studies should address the question of the role of static indexes in predicting cardiac output improvement following fluid infusion in spontaneously breathing patients. PMID- 16352958 TI - Clinical implications of temperature curve complexity in critically ill patients. AB - OBJECTIVE: In certain physiologic systems, disease is associated with a loss of complexity in system's output. We test the hypothesis that, in critically ill patients, there is an inverse relation between the complexity of the temperature curve and the clinical status. We also consider whether complexity analysis of the temperature curve may have prognostic value. DESIGN: Prospective, observational study. SETTING: Intensive care unit of a general hospital in Madrid, Spain. PATIENTS: Twenty-four successive patients admitted in the intensive care unit with multiple organ failure. INTERVENTIONS: Skin temperature was measured every 10 mins from inclusion in the study until discharge or death (median length of stay 18.8 days, interquartile range 86). MEASUREMENTS: From the temperature time series, hourly approximate entropy measurements were obtained. Clinical status was evaluated using the Sequential Organ Failure Assessment (SOFA) score. MAIN RESULTS: A significant inverse relationship between approximate entropy and the attributed SOFA score was observed in 89% of the patients considered. Both mean and minimum approximate entropy were significantly lower in patients who died than in patients who survived (mean approximate entropy, 0.47 vs. 0.61; minimum approximate entropy, 0.24 vs. 0.40; in both cases p < .001). To evaluate the prognostic value of both mean and minimum approximate entropy, we fitted logistic regression models against survival. An increase in 0.1 units in minimum or mean approximate entropy increased 15.4- and 18.5-fold the odds of surviving, respectively. CONCLUSIONS: The clinical status of patients suffering multiple organ failure is inversely correlated to the complexity of the temperature curve expressed as approximate entropy. Reduced complexity has dismal prognostic implications. Its assessment is noninvasive and inexpensive and allows for real-time continuous monitoring of clinical status. PMID- 16352959 TI - Impact of admission hyperglycemia on hospital mortality in various intensive care unit populations. AB - OBJECTIVE: Hyperglycemia in intensive care unit patients has been associated with an increased mortality rate, and institutions have already begun tight glucose control programs based on a limited number of clinical trials in restricted populations. This study aimed to assess the generalizability of the association between hyperglycemia and in-hospital mortality in different intensive care unit types adjusting for illness severity and diabetic history. DESIGN: Retrospective cohort study. SETTING: The medical, cardiothoracic surgery, cardiac, general surgical, and neurosurgical intensive care units of the University of Maryland Medical Center. PATIENTS: Patients admitted between July 1996 and January 1998 with length of stay > or = 24 hrs (n = 2713). INTERVENTIONS: On intensive care unit admission, blood glucose and other physiologic variables were evaluated. Regular measurements were taken for calculation of Acute Physiology and Chronic Health Evaluation III scoring. Patients were followed through hospital discharge. Admission blood glucose was used to classify patients as hyperglycemic (> 200 mg/dL) or normoglycemic (60-200 mg/dL). The contribution of hyperglycemia to in hospital mortality stratified by intensive care unit type and diabetes history while controlling for illness severity was estimated by logistic regression. MEASUREMENTS AND MAIN RESULTS: The adjusted odds ratios for death comparing all patients with hyperglycemia to those without were 0.81 (95% confidence interval, 0.37, 1.77) and 1.76 (95% confidence interval, 1.23, 2.53) for those with and without diabetic history, respectively. Higher mortality was seen in hyperglycemic patients without diabetic history in the cardiothoracic, (adjusted odds ratio, 2.84 [1.21, 6.63]), cardiac (adjusted odds ratio, 2.64 [1.14, 6.10]), and neurosurgical units (adjusted odds ratio, 2.96 [1.51, 5.77]) but not the medical or surgical intensive care units or in patients with diabetic history. CONCLUSIONS: The association between hyperglycemia on intensive care unit admission and in-hospital mortality was not uniform in the study population; hyperglycemia was an independent risk factor only in patients without diabetic history in the cardiac, cardiothoracic, and neurosurgical intensive care units. PMID- 16352960 TI - Reliability of point-of-care testing for glucose measurement in critically ill adults. AB - BACKGROUND: Glycemic control is increasingly being recognized as a priority in the treatment of critically ill patients. Titration and monitoring of insulin infusions involve frequent blood glucose measurement to achieve target glucose ranges and prevent adverse events related to hypoglycemia. Therefore, it is imperative that bedside glucose testing methods be safe and accurate. OBJECTIVE: To determine the accuracy and clinical impact of three common methods of bedside point-of-care testing for glucose measurements in critically ill patients receiving insulin infusions. DESIGN: Prospective observational study. SETTING: A 21-bed mixed medical/surgical intensive care unit of a tertiary care teaching hospital. PATIENTS: Thirty consecutive critically ill patients who were vasopressor-dependent (n = 10), had significant peripheral edema (n = 10), or were admitted following major surgery (n = 10). MEASUREMENTS: Findings from three different methods of glucose measurement were compared with central laboratory measurements: (1) glucose meter analysis of capillary blood (fingerstick); (2) glucose meter analysis of arterial blood; and (3) blood gas/chemistry analysis of arterial blood. Patients were enrolled for a maximum of 3 days and had a maximum of nine sets of measurements determined during this time. RESULTS: Clinical agreement with the central laboratory was significantly better with arterial blood analysis (69.9% and 76.5% for glucose meter and blood gas/chemistry analysis, respectively) than with capillary blood analysis (56.8%; p = .039 and .001, respectively). During hypoglycemia, clinical agreement was only 26.3% with capillary blood analysis and 55.6% and 64.9% for glucose meter and blood gas/chemistry analysis of arterial blood (p = .010 and <.001, respectively). Glucose meter analysis of both arterial and capillary blood tended to provide higher glucose values, whereas blood gas/chemistry analysis of arterial blood tended to yield lower glucose values. CONCLUSIONS: The magnitude of the differences in the glucose values offered by the four different methods of glucose measurement led to frequent clinical disagreements regarding insulin dose titration in the context of an insulin infusion protocol for aggressive glucose control. PMID- 16352961 TI - Outcome in elderly patients with severe infection is influenced by sex hormones but not gender. AB - OBJECTIVE: The influence of gender as a prognostic variable in patients with severe infections is still controversial. Sex steroid hormones have an important impact on the immune system and vice versa, and prospective studies on the hormonal changes during severe infection are lacking. The objective was to compare the influences of gender and adrenal sex steroid hormone levels on hospital mortality rate in patients with infections. DESIGN: Prospective observational study conducted between January 1995 and December 2000. SETTING: University-based level I intensive care unit. PATIENTS: Included were 208 males and 100 females with severe infection at admission to the ICU. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Mortality rate during hospitalization was analyzed for correlation to gender and the levels of testosterone, 17beta estradiol, and progesterone; source and clinical signs of infection; Acute Physiology and Chronic Health Evaluation II score; or age. There were no differences in demographic or infectious characteristics between males and females; the survival rate was similar. Males had significantly reduced testosterone levels. Elevation of the steroid hormones 17beta-estradiol (1.5 fold), progesterone (5-fold), and cortisol (1.5-fold) occurred in both genders to the same extent. In addition, testosterone was elevated in septic females and correlated with 17beta-estradiol. Nonsurvivors of both genders had significantly elevated 17beta-estradiol levels. Progesterone was particularly high in nonsurviving males, whereas testosterone was elevated in nonsurviving females. Mortality rate was correlated with high 17beta-estradiol and progesterone in males but with 17beta-estradiol and testosterone in females. Cortisol or dehydroepiandrostenedione sulfate levels were not associated with mortality rates. CONCLUSIONS: In elderly patients with infections, mortality was not dependent on gender but was correlated with elevated 17beta-estradiol in both genders, with elevated progesterone in males and elevated testosterone in females. Although the latter sex hormones may derive from the adrenals, cortisol levels were only moderately increased and not associated with survival. The high 17beta-estradiol concentrations implicate an increased aromatase activity. Therefore, other pathways of sex steroid production must be involved. PMID- 16352962 TI - Nitric oxide donors alter cardiomyocyte cytokine secretion and cardiac function. AB - OBJECTS: The mechanisms by which nitric oxide produces beneficial/detrimental effects on physiologic function are unclear. In this study, we hypothesized that nitric oxide promotes cyclic guanosine monophosphate (cGMP) formation, which, in turn, promotes cardiomyocyte secretion of inflammatory cytokines as well as accumulation of intracellular Na+/Ca2+; these factors contribute to altered cardiac contractile function. DESIGN: Laboratory study. SETTING: Medical Center. SUBJECTS: Adult Sprague Dawley rats weighing 325-350 g. INTERVENTIONS: Cardiomyocytes were prepared by collagenase perfusion of rat hearts; cells were plated (5 x 10(4) cells/microtiter well) and challenged with either vehicle or nitric oxide donor (S-nitroso-N-acetyl-penicillamine [SNAP] or PAPA NONOATE, 3-[2 Hydroxy-2-nitroso-1-propythdrazinol]-1-propanamine], NOC-15 [PAPA-NO], 0.3 or 1.0 mM of each nitric oxide donor) in the presence/absence of methylene blue (10 microM/L to inhibit cGMP). After 3 hrs, supernatants were collected to measure nitrite/nitrate (nitric oxide), cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin-6), and cGMP levels; cells were then loaded with a fluorescent indicator (Fura-2AM or sodium-binding benzofurzan isophthalate) to measure myocyte Ca2+ or Na+, respectively. Parallel experiments included the addition of nitric oxide donor (0.3 or 1.0 mM SNAP or PAPA-NO) to perfused hearts in presence or absence of the methylene blue to examine cGMP-mediated effects on myocardial contraction-relaxation, while other experiments determined a) potential lipopolysaccharide contamination of myocyte preparations; and b) whether a cGMP analogue recapitulated the effects of nitric oxide donors on cytokine secretion. MEASUREMENTS AND MAIN RESULTS: Nitric oxide donors produced a dose-dependent increase in cGMP levels in myocyte supernatants as well as an increase in myocyte cytokine secretion, increased myocyte loading of Na+/Ca2+, and produced myocardial contractile dysfunction. Addition of the cGMP analog, 8 bromo-cGMP, recapitulated the effects of nitric oxide donors on myocyte cytokine secretion. Nitric oxide donor-related effects were ablated by pretreatment of myocytes or isolated hearts with methylene blue. Treatment of myocytes with recombinant bactericidal/permeability-increasing protein to scavenge lipopolysaccharide confirmed that cytokine responses to nitric oxide donors were not related to lipopolysaccharide contamination of myocyte preparations. CONCLUSIONS: We suggest that nitric oxide synthesis in injury and disease promotes cGMP formation, which, in turn, modulates cardiac contraction/relaxation by a) altering cardiomyocyte secretion of inflammatory cytokines and b) altering myocyte handling of Na+/Ca2+. PMID- 16352963 TI - Intermittent spontaneous breathing protects the rat diaphragm from mechanical ventilation effects. AB - OBJECTIVE: Short-term mechanical ventilation has been proven to reduce diaphragm force and fiber dimensions. We hypothesized that intermittent spontaneous breathing during the course of mechanical ventilation would minimize the effects of mechanical ventilation on diaphragm force and expression levels of transcription factors (MyoD and myogenin). DESIGN: Randomized, controlled experiment. SETTING: Animal basic science laboratory. SUBJECTS: Male Wistar rats, weighing 350-500 g. INTERVENTIONS: Anesthetized and tracheotomized rats were submitted to either 24 hrs of spontaneous breathing (SB, n = 5), 24 hrs of continuous controlled mechanical ventilation (CMV, n = 7), or controlled mechanical ventilation with intermittent spontaneous breathing: 60 mins every 5 hrs of mechanical ventilation repeated four times (ISB60, n = 8) or 5 mins every 5 hrs 55 mins of mechanical ventilation repeated four times (SB5, n = 9). They were compared with control animals free from intervention (C, n = 5). MEASUREMENTS AND MAIN RESULTS: The profile of the diaphragm force-frequency curve of the controls and SB group was significantly different from that of the ISB and CMV groups; especially, the mean asymptotic force was less in the ISB and CMV compared with controls and SB. CMV resulted in a significant decrease in the diaphragm type I (-26%, p < .05 vs. C) and type IIx/b (-39%, p < .005 vs. C and SB) cross-sectional area, whereas this was not observed in the ISB groups. Diaphragm MyoD protein expression was significantly decreased after ISB60 (-35%, p < .0001 vs. C and SB) and even more after CMV (-73%, p < .0001 vs. others). The same pattern was observed with myogenin protein levels. Positive relationships between diaphragm MyoD and myogenin protein levels and diaphragm force were observed. CONCLUSIONS: The data demonstrated that intermittent spontaneous breathing during the course of mechanical ventilation may minimize the deleterious effect of controlled mechanical ventilation on diaphragm force, fiber dimensions, and expression of transcription factors. PMID- 16352964 TI - Adrenomedullin treatment abolishes ileal mucosal hypoperfusion induced by Staphylococcus aureus alpha-toxin--an intravital microscopic study on an isolated rat ileum. AB - OBJECTIVE: Disturbances of intestinal microcirculation associated with sepsis and septic shock result in diminished mucosal oxygenation. Tissue hypoxia as well as mediator formation may lead to intestinal mucosa dysfunction. As a consequence, bacteria and their products as well as gut-derived inflammatory mediators may further perpetuate septic and inflammatory events. Adrenomedullin is produced in the mucosa of the gastrointestinal tract and has been shown to improve survival in experimental sepsis. Using pore-forming Staphylococcus aureus alpha-toxin as a potent initiator of inflammatory reactions, we tested the hypothesis that exogenously added adrenomedullin improves ileal mucosal perfusion. DESIGN: Prospective, experimental study. SETTING: University laboratory. SUBJECTS: Isolated perfused ileum from male Sprague-Dawley rats INTERVENTIONS: Adrenomedullin treatment of S. aureus alpha-toxin infused ileum. MEASUREMENT AND MAIN RESULTS: An infusion of alpha-toxin (0.05 microg/mL) induced a significant decrease of red blood cell velocity in villus terminal arterioles from 1.7 to 0.7 mm/sec assessed by intravital microscopy. This was accompanied by a significant reduction of mucosal hemoglobin oxygenation from 71.8% to 17.5% and impaired oxygen uptake. At constant bulk flow and oxygen delivery, these data indicate a redistribution of blood perfusion away from mucosa. Subsequent intervention with 0.1 microM adrenomedullin redistributed blood flow back toward the mucosa, causing an improvement of mucosal hemoglobin oxygenation and of organ oxygen uptake. CONCLUSION: These data suggest that exogenously added adrenomedullin protects ileum mucosa by diminishing alpha-toxin-induced microcirculatory disturbances. Further investigations will have to clarify the therapeutic potential of adrenomedullin in sepsis-related gut dysfunction. PMID- 16352966 TI - In-hospital cardiac arrest. AB - To review the current management of in-hospital cardiac arrest and to identify variables that influence outcomes, OLDMEDLINE from 1950 to 1966 and MEDLINE from 1966 to March 2005 were searched using the keywords cardiopulmonary resuscitation, cardiac arrest, in hospital, and adult. Secondary sources were derived from review publications and personal communications by one of the authors. There is no secure evidence that the ultimate outcomes after cardiopulmonary resuscitation in settings of in-hospital cardiac arrest have improved in the >40 yrs that followed the landmark report by Kouwenhoven, Jude, and Knickerbocker, which launched the modern era of cardiopulmonary resuscitation. A paucity of objective measurements preclude secure protocols for sequencing of interventions and, even more, when to initiate and discontinue cardiopulmonary resuscitation. The preparedness of both physicians and nursing professionals to implement the published guidelines has itself been questioned. Whereas early access defibrillation with automated external defibrillators may be of benefit in out-of-hospital settings, there has as yet been no secure evidence that automated external defibrillators have had a favorable impact on in-hospital cardiopulmonary resuscitation when used on infrequent occasions by first responders. This contrasts with the much greater success of advanced life support providers and especially when electrical defibrillation is promptly performed by expertly trained personnel after onset of cardiac arrest. Outcomes are therefore improved in critical care settings and especially in coronary care units in which patients are continuously monitored. PMID- 16352965 TI - Central nervous system complications during pediatric extracorporeal life support: incidence and risk factors. AB - OBJECTIVE: Identify the incidence and risk factors for development of acute, severe central nervous system (CNS) complications of pediatric extracorporeal life support (ECLS). DESIGN: Retrospective review of Extracorporeal Life Support Organization (ELSO) registry database. SETTING: Pediatric intensive care units of 115 tertiary centers internationally. PATIENTS: Pediatric patients, 1 month to 18 yrs of age, who had ECLS between the years 1981-2002. MEASUREMENTS AND MAIN RESULTS: Data concerning 4,942 patients who underwent one run of ECLS were analyzed. Six hundred thirty-six patients (12.9%) developed acute, severe CNS complications. Patients who required ECLS during extracorporeal cardiopulmonary resuscitation (n = 161; 3.3%) were more likely to develop CNS complications (n = 42; 26.1%) than patients who did not have extracorporeal cardiopulmonary resuscitation (p < .001; odds ratio [OR], 2.48; 95% confidence interval [CI], 1.73-3.57). Stepwise logistic regression analysis of therapies patients received before initiation of ECLS showed that the use of a left ventricular assist device (p = .001; OR, 3.45; 95% CI, 1.64-7.22), bicarbonate (p < .001; OR, 1.61; 95% CI, 1.26-2.05), and vasopressor/inotropic medications (p = .035; OR, 1.22; 95% CI, 1.01-1.48) were significant independent predictors of development of CNS complications. Among patients who had pulmonary failure as an indication for ECLS, the CNS complication rate was significantly higher for those treated with venoarterial ECLS than those who had venovenous ECLS (13.5% vs. 5.7%; p < .001; OR, 0.43; 95% CI, 0.34-0.67). Multiple logistic regression analysis of the complications other than CNS complications associated with the use of ECLS showed that pH <7.20, creatinine concentration >3.0 mg/dL, use of inotropes, presence of myocardial stun, and requirement of cardiopulmonary resuscitation during ECLS independently predicted development of CNS complications. CONCLUSION: Patients who have metabolic acidosis, a bicarbonate or inotrope/vasopressor requirement, cardiopulmonary resuscitation, or a left ventricular assist device before initiation of ECLS are at greater risk for development of CNS complications. After initiation of ECLS, patients who develop renal failure or metabolic acidosis or undergo venoarterial ECLS should be closely monitored for development of CNS complications. PMID- 16352967 TI - Both percentage of gammadelta T lymphocytes and CD3 expression are reduced during septic shock. AB - OBJECTIVE: The mechanisms involved during sepsis-induced immunosuppression are far from being extensively established. The objective of the present study was to investigate whether two characteristics of T cells were altered in this situation: the percentage of circulating gammadelta T lymphocytes and the level of CD3 expression on T lymphocytes. DESIGN: Observational study. SETTING: Adult intensive care units in a university hospital. PATIENTS: Patients with septic shock (n = 21) and healthy individuals (n = 21). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: In patients, we first observed the decreased percentage of gammadelta T lymphocytes in peripheral blood (1% [0.7-3.1], median [interquartile range]) in comparison with healthy individuals (3.5% [2.1-4.8]). Regarding CD3, we measured a highly significant decrease of its expression on both alphabeta and gammadelta T lymphocytes from patients (p < .005), whereas the CD3 mean fluorescence intensities ratio (gammadelta/alphabeta) was not affected: 2.2 [2.1-2.4] and 2.1 [1.9-2.3] in healthy individuals and septic patients, respectively. The magnitude in the decrease of CD3 expression was thus similar in alphabeta and gammadelta cells, suggesting a common down-regulation mechanism for both T-cell lineages. CONCLUSIONS: Combined with a reduced percentage of monocytes expressing human leukocyte antigen-DR, a reduced CD3 expression may be involved in the failure of antigen presentation depicted after septic shock, whereas the diminished percentage of circulating gammadelta T cells could be partly responsible for the elevated incidence of secondary infections. These two observations constitute additional pieces of the complex puzzle of sepsis-induced immunosuppression. PMID- 16352968 TI - The compassionate clinician: attending to the spiritual needs of self and others. PMID- 16352969 TI - The swinging pendulum of corticosteroid use in the intensive care unit: has it swung too far or not far enough? PMID- 16352970 TI - Refrigerated intravenous fluids: kick-starting the cooling process. PMID- 16352971 TI - Reversible myocardial dysfunction in sepsis and ischemia. PMID- 16352972 TI - Admission hyperglycemia and outcome: the ongoing story. PMID- 16352973 TI - How accurate are currently used methods of determining glycemia in critically ill patients, and do they affect their clinical course? PMID- 16352974 TI - Nitric oxide and inflammatory cytokines in the heart: the presence of positive feedback loops. PMID- 16352975 TI - Ventilator-induced diaphragmatic dysfunction: keep working. PMID- 16352976 TI - Pediatric extracorporeal life support and central nervous system injury. PMID- 16352978 TI - Does red blood cell transfusion increase local cerebral oxygenation? PMID- 16352979 TI - Transfusion and mechanical ventilation: two interrelated causes of acute lung injury? PMID- 16352980 TI - Intensivists should use bedside echocardiography. PMID- 16352983 TI - In patients with obstructive pulmonary disease during controlled ventilation, PEEP decreases dynamic hyperinflation: is this response really "paradoxical"? PMID- 16352985 TI - [Short-term results of radical surgeries for non-small cell cancer of the lung]. AB - The results of radical surgical treatment of 358 patients with NSCCL were analyzed. Majority of patients were men (84.6%) and persons older 60 years -- 239 (66.8%) patients. Central cancer was diagnosed in 164 (45,8%) patients, peripheral -- in 194 (54.2%). There were 137 (38.3%) pneumonectomies, 184 (51.4%) lobectomies; 149 (41.6%) surgeries were combined. Complications after radical surgeries were seen in 92 (25.7%) of 358 patients, including 18 (5.0%) patients with <> complications and 74 (21%) patients with <> ones. Complications were seen more often in patients over 60 years and occurred less frequently after segmentectomy (7 patients). Lethality was 5.0% (18 patients). Respiratory complications (7 cases) and pulmonary embolism (5 patients) were the main causes of lethal outcome. PMID- 16352986 TI - [Surgical treatment of patients with rupture of abdominal aorto-aneurysm]. AB - Thr experience in surgical treatment of 69 patients with rupture of abdominal aorto-aneurysm was analyzed. Multifactorial statistical analysis was carried out to determine detect influence of different parameters on postoperative lethality. The most significant parameters were initial blood loss (odds ratio (OR) -- 10.4, confidence interval (CI) -- 2.0-37.1, intraoperative hemorrhage (OR=8.5, CI 1.5 27.8), minute diuresis (OR=15,7, CI 1.8- 49.3), location of rupture (OR=12.1, CI 2.0-33.5). Combination of two and more factors led to the most unfavorable prognosis. The proposed algorithm allowed the authors to optimize treatment policy and improve significantly results of surgical treatment of patients with rupture of abdominal aorto-aneurysm: postoperative lethality has decreased from 71.4 to 28.5%. PMID- 16352987 TI - [Short-term results of "large" amputations in patients with chronic lower limb arterial occlusions]. AB - The results of examinations and treatment are presented for 238 patients with lower limb arterial occlusions after "large" amputation for critical limb ischemia. Arterial reconstructions and lumbar sympathectomy, adequate choice of amputation level based on objective parameters of macro- and microcirculation, epidural anesthesia, prophylaxis of early thromboembolism and cardiovascular complications are measures which can improve results of "large" amputations in patients with lower limb arterial occlusions. PMID- 16352988 TI - [Disputable problems in diagnosis and treatment of catecholamine-secreting tumors]. AB - The article covers the main problems and disputable points in diagnosis and treatment of pheochromocytoma: differential diagnosis before surgery and treatment which policy. The authors define a specific group of patients required a comprehensive deep examination (group of risk) for chromaffin tissue. Differentiated use of different methods of diagnosis of catecholamine-secreting tumors is validated. Some last est negative tendencies in adrenal surgery are also discussed. PMID- 16352989 TI - [Diagnosis and differential treatment of acute pancreatitis of biliary etiology]. AB - The experience in examination and treatment of 208 patients with acute pancreatitis of biliary etiology was analyzed. Complex endoscopic treatment was carried out in 88% patients. If there are indications, it is reasonable to perform retrograde pancreatocholangiography (RPCG) and endoscopic papillosphincterotomy (EPST) during surgery in patients with concomitant enzymatic ascites-peritonitis. In the others patients RPCG and EPST must be regarded as the first stage of treatment. Surgical procedure of choice at the second stage of treatment is laparoscopic cholecystectomy. PMID- 16352990 TI - [Total duodenopancreatosplenectomy in surgery of pancreatic and gastric cancer]. AB - The experience of treatment of 23 patients undergone total duodenopancreatosplenectomy was analyzed. Postoperative complications (mainly endocrine pathology) were seen in 21 patients. There were no cases of anastomoses insufficiency. Seven patients died. In group of patients who underwent combined gastrectomy 2 of 5 persons died. There were no lethal outcomes after angiosurgical procedures (5 patients). PMID- 16352991 TI - [Extracorporeal detoxication in patients with destructive pancreatitis]. AB - Criteria of severity of endogenous intoxication in patients with destructive pancreatitis in enzymatic toxemia are discussed. It is demonstrated that the level of middle mass molecules and integral index of affection of the pancreas, abdominal cavity and retroperitoneal space are objective criteria of endogenous intoxication and can be used for determination of indications for extracorporeal detoxication and for assessment of treatment efficacy. Prescription of extracorporeal detoxication according to these criteria in an early period of the disease before multiple organ failure development promotes faster management of endotoxemia, prophylaxis of septic complications, decreases lethality and period of treatment of patients with destructive pancreatitis. PMID- 16352992 TI - [Method of cover of "difficult" duodenal stump in resection of the stomach]. AB - The experience with 233 stomach resections in "difficult" ulcers is analyzed. The main cause of complications and lethality after these surgeries is duodenal stump sutures insufficiency seen in 17 (7.3%) patients, and in 10 (4.3%) patients it was the cause of death. Original duodenal stump cover was performed in 86 patients. There were no cases of suture insufficiency, two patients died of pneumonia and hepatic failure. PMID- 16352993 TI - [Surgical treatment of stage IV cardial achalasia]. AB - An original method of surgical treatment of patients with stage IV cardial achalasia was developed. Seven patients aged 42 to 59 years underwent surgery according this method. Objective methods of examination and life quality study with GIQLI carried out 3 months to 5 years after surgery demonstrate satisfactory anatomic and functional results. PMID- 16352994 TI - [Catheterization of the external jugular vein for hemodialysis]. AB - Permanent central catheter for hemodialysis was inserted via external jugular vein in 95 patients. In all the cases the catheter blood flow was more than 250 ml/min. There were no severe complications. The procedure of insertion of permanent catheter for hemodialysis via the external jugular vein is described. It is concluded that the insertion of permanent catheter for hemodialysis via the external jugular vein is a simple and short-time procedure without any severe complications. PMID- 16352995 TI - [Erysipelas as the cause of lymphedema]. AB - The results of the treatment of 237 patients with lymphedema and recurrent erysipelas are analyzed. Direct correlation between the number of erysipelas in anamnesis and degree of lymphedema, and also their reciprocal aggravation are demonstrated that permits us to regard erysipelas as the cause of lymphedema. PMID- 16352996 TI - [Pathogenesis and treatment of inguinal hernias]. AB - The experience in surgical treatment of 402 men with inguinal hernias according to the original method (aponeuroplasty of posterior wall of inguinal canal "without tension") is analyzed. Long-term results were evaluated in 374 patients. There were no recurrences of hernias. Problems of pathogenesis of inguinal hernias are discussed. PMID- 16352997 TI - [Foreign bodies of the rectum]. AB - The experience in the treatment of 143 patients with foreign bodies of the rectum and sigmoid colon is analyzed. Groups of foreign bodies are distinguished according to their size, structure and ways of occurring. Methods of extraction of foreign bodies in typical and non-typical situations are described. It is demonstrated that in 2.8% cases of extraction surgeries laparotomy is required even without perforation of this organ. PMID- 16352998 TI - [Method of forming of compressive terminal colon anastomosis]. AB - The method of creation of compressive terminal colon anastomosis with implants of TN-10 titanium nikelid was developed. Experimental studies carried out on 60 mongrel dogs demonstrated higher physical strength and biological containment of these anastomoses compared with manual and mechanical ones. Regeneration completed earlier after surgery and was accompanied by less inflammation. Clinical study was carried out in 54 patients. There were no lethal outcomes, insufficiency of anastomosis was seen in 1 (1.9%) case. PMID- 16352999 TI - [Problem of ENT hospital infection]. AB - Five-year monitoring of microbial environment of the ENT department stated its relative qualitative persistence. Circulating in hospitals microbial strains have a direct influence on the course of the infectious process in the patients. Hospital infection in the ENT department replaces the initial pathogen for a polyresistant hospital pathogen or forms a persistent microbial association. PMID- 16353000 TI - [Viral diseases as a predisposing factor of developing secondary oto- and rhinogenic bacterial meningitis]. AB - Effects of viruses and bacterial associations are suggested to promote secondary purulent intracranial complications in ENT patients. Fundamental aspects of pathogenesis of viral neuroinfections and pathogenetic mechanisms of different conditions provoked by viral factors promoting bacterial complications are presented. Hemoculture, and cerebrospinal fluid in 24 patients with secondary purulent rhinogenic and otogenic meningitis were studied with polymerase chain reaction for enteroviruses, cytomegaloviruses, Epstein-Barr, herpes, zoster viruses. The viruses were present in the studied media in 29.4% samples. PMID- 16353001 TI - [Domestic 1.44 mcm Nd:YAG laser in combined treatment of dacryocystitis complicated by abscess formation]. AB - We have treated 12 patients with acute dacryocystitis complicated by abscess formation. All the patients were examined by an ophthalmologist and otolaryngologist. The patients received intravenous antibiotics followed by Nd:YAG laser dacryocystorhinostomy via transcanalicular approach under the endonasal endoscopic control. The treatment resulted in resolution of symptoms and signs of acute dacryocystitis complicated by abscess formation in all the patients. No recurrence of acute dacryocystitis occurred during the median follow up period of 11 months. Ostium patency defined as the absence of epiphora and observation of irrigated lacrimal fluorescein at the ostium was achieved in 9 patients. Epiphora recurred in 3 cases. Use of Nd:YAG laser reduces the duration of the operation and provides maximal functional and cosmetic results. PMID- 16353002 TI - [Grounds for treatment of patients with infectious-allergic rhinitis with bacterial allergens]. AB - We studied mechanisms of cell and humoral immunity in the course of allergen specific immunotherapy (ASIT) with bacterial allergens in patients with infectious-allergic rhinosinusitis (IAR). Measurements of cytokines determining the type of immune response in conduction of ASIT with bacterial allergens (IL-2 and IL-6) have demonstrated that ASIT shifted cytokines balance to prevalence of Th1 cells. Good and excellent results were achieved in 80% IAR patients. Thus, T lymphocyte activation of immune system is essential in the mechanism of IAR formation. PMID- 16353003 TI - [Psychosomatic status of patients with cochleovestibular disorders. Correction methods]. AB - Psychosomatic status was studied in 106 patients aged 18-65 years with acute and chronic cochleovestibular disorders. Hearing problems, noise in the ears and vestibular disorders have a negative psychogenic effect in patients with cochleovestibulopathy. The majority of the patients need psychological and psychopharmacological care to relieve anxiety and form adequate attitude to the disease. Correction of psychosomatic disorders raises efficacy of the treatment of cochleovestibular diseases. PMID- 16353004 TI - [Audiological diagnosis of cochleovestibular disorders caused by internal ear hydrops]. AB - A total of 75 patients with clinical signs of internal ear hydrops (IEH) were examined using dehydration tests and electrocochleography. The patients were divided into groups by the results of pure tone threshold audiometry according to the international classification. Both methods proved highly informative. Positive dehydration tests were obtained in 57% cases, positive electrocochleography evidence was observed in 73% cases. An increased SP/AP amplitude was seen in 65% cases. Both methods allowed IEH detection in 82% cases. In normal hearing and hypoacusis of the first degree electrocochleography is more effective than dehydration tests in detection of IEH, but with hearing loss deterioration this advantage lessens. It is shown that patients can suffer from IEH in normal values of pure tone threshold audiometry. This should be taken into consideration in examination of patients who do not complain of hypoacusis. PMID- 16353005 TI - [Establishment of hearing defects relation with occupation in civil aviation crews]. AB - Occupational neurosensory hypoacusis is essential in the pattern of ENT morbidity in civil aviation crews. How to establish relations between hearing problems and occupation in civil aviation is shown. The algorithm of occupational neurosensory hypoacusis diagnosis in civil flights crews is proposed. PMID- 16353006 TI - [Treatment of patients with chronic adenoiditis and abnormal coagulation]. AB - Chronic adenoiditis (CA) prevails among ENT diseases in children. It often demands surgical intervention. Frequent hemorrhagic complications during adenotomy necessitate assessment of the patient's hemostasis. A preoperative examination of 43 children has detected primarily dysaggregation thrombocytopathies. Streptococcus pneumoniae is a dominating etiological agent. Chronic focal infection is involved in pathogenesis of systemic disorders in CA including those provoking coagulatory disturbances. An algorithm of hemostatic disorders diagnosis in CA children is proposed as well as an effective program of targeted pre- and intraoperative hemostatic therapy. PMID- 16353007 TI - [Clinicofunctional parallels in posttraumatic deformities of the nasal tip]. AB - The analysis of different posttraumatic deformities of the nasal tip gave evidence for a close correlation of internal nose deformities with nasal breathing ranging from common changes in the internal nose (hypertrophy of the inferior turbinated bone) to atresia when nasal breathing is completely blocked. Because of tissue edema, respiration improved and normalized only 1 and 3 months after surgery. It is concluded that functional examinations before and after surgery are beneficial for patients and warrant optimal terms of surgery. PMID- 16353008 TI - [Functionally sparing operation in locally advanced laryngeal cancer]. AB - The proposed reconstructive operation in patients with locally advanced laryngeal cancer. The operation implies removal of the tumor-affected half of the larynx with reconstruction of the lost structures. The resected upper corniculum of the thyroid cartilage is preserved and sutured to the stump of the cricoid cartilage seal. The lateral laryngeal wall is created from cervical skin. The proposed technique was used in surgery of 30 patients. They recovered swallowing, respiratory and speech functions. PMID- 16353009 TI - [Etiotropic therapy with sparfloxacine in ENT practice]. PMID- 16353010 TI - [Vocal problems in children and adolescents]. AB - The examination of 2000 children and adolescents diagnosed vocal problems in the form of functional and organic dysphonias in 3.5% cases. Mutational, hypotonic dysphonias occurred more frequently among functional diseases; nodules of the vocal cords, acute and chronic laryngitis -- among organic diseases. Causes of their development and therapeutic policy are specified. PMID- 16353011 TI - [Efficacy of betaserk in experimental motor disease]. PMID- 16353012 TI - [Optimal outpatient treatment of maxillary inflammation]. AB - To improve treatment of outpatients with maxillary inflammation (MI) it is recommended, in case of failure of previous standard conservative therapy, to drain the maxillary sinus for a long time. Catheter should be inserted into the maxillary sinus for maximum time but not longer than for 10-12 days. When inflammation is not corrected by the first course of therapy, the drainage is removed and the patients receive conditioning, mucolytic and immunostimulating therapy in combination with physiotherapy for 8-10 days with a following another course of drainage. In persistent maxillary sinusitis, it is possible to use long term lavage of the maxillary sinus via catheter attached to the dispenser. Such an approach is effective in the majority of patients. PMID- 16353013 TI - [The technique of endaural drainage of the antrum]. AB - Antrodrainage methods are reviewed. To easy endaural drainage, a special drill is proposed. It has been used for trephine puncture of the antrum in 60 patients (55 with otitis purulenta chronica and in 5 with exudative otitis media). Positive diagnostic and therapeutic significance of endaural drainage as well as necessity of its further perfection and wider usage in practice are emphasized. PMID- 16353014 TI - [Laryngeal carcinoid]. PMID- 16353015 TI - [A complicated course of juvenile respiratory papillomatosis]. PMID- 16353016 TI - [Multiple skin cylindroma of both floors of the auricle and internal acoustic meatus]. PMID- 16353017 TI - [Present-day aspects of surgical treatment of mitral restenosis]. AB - Tactical and technical features of treatment of patients with mitral restenosis are discussed. The results (surgical lethality was 6.41%) confirm the necessity of surgical treatment of mitral restenosis with artificial circulation. Close commissurotomy may be regarded as the stage of the treatment of mitral stenosis, particularly under real existing Russian conditions. PMID- 16353018 TI - [Role of reflux in evolution and prognosis of acute thrombophlebitis of lower extremities varicose veins]. AB - Role of venous reflux in evolution and prediction of acute varicotrombophlebitis (AVTP) has been evaluated with coloured duplex scanning performed as daily monitoring. It is demonstrated that venous reflux determines the direction and rate of thromb formation, it potential of embolism, promotes formation of floating thrombi. Study of venous reflux permits to predict the evolution of AVTP. It is one of the key elements which influences the thrombus formation and determines policy of treatment of varicose disease complicated with AVTP. PMID- 16353019 TI - [Choice of the extent and the terms of sanifying procedures after vascular reconstructive surgeries in patients with stage IV chronic arterial lower limb insufficiency]. AB - Results of vascular reconstructions with the following sanifying procedures performed in 82 patients with stage IV chronic arterial lower limb insufficiency are analyzed. Based on complex evaluation of microcirculation three types of tissues ischemic lesion are revealed: irreversible, severe and mild. Patients with irreversible lesion of the tissues of a distal part of the lower extremity require vascular reconstruction with simultaneous amputation at the upper third of the shank. In a severe reversible ischemic lesion of the tissues radical sanifying surgery with preservation of limb support function is better to perform not earlier than one month after vascular reconstruction. Mild ischemia of tissues permits performing minimal sanifying procedure with primary closure of the wound simultaneously with vascular surgery. PMID- 16353020 TI - [Diagnosis and surgical treatment of aneurysms of the popliteal artery]. AB - Nineteen patients with aneurysms of the popliteal artery have been operated. Indications and type of surgery were determined on the basis of the clinical picture, angiography and duplex scanning data. There were 19 surgeries: ligation of afferent and efferent vessels of aneurysm with auto-venous grafting (8), resection of aneurysm with auto-venous grafting (5), dissection of aneurysm with auto-venous grafting and coverage with duplication from aneurysmatic walls (3), endovascular grafting of aneurysm (2), ligation of afferent vessel of aneurysm (1). In early postoperative period after open surgical revascularizations thrombosis was seen in 4 (25%) patients. One patient died on day 20 after surgery due to decompensation of severe concomitant disease. The patient who has undergone ligation of afferent vessel without revascularization has discharged after compensation of acute ischemia. PMID- 16353021 TI - [Diagnosis and results of surgical treatment of rare giant intrathoracic tumors]. AB - Surgical treatment was performed in 19 patients aged 30 to 64 years with rare giant intrathoracic tumors. The tumor located in the mediastinum in 8 patients, in the lungs -- in 6, in the chest wall with intrathoracic growth -- in 5 patients. Diagnostic examinations included x-ray (including CT) and morphological methods. Sarcoma was diagnosed in 10 patients, mesenchymal tumors -- in 4, neurogenic tumors -- in 3, thymoma -- in 2 patients. Radical surgeries were performed in 15 patients, palliative -- in 3, explorative mini-thoracotomy -- in 1. Computed tomography and CT-assisted puncture biopsy improve the quality of preoperative diagnosis. Radical surgery is the method of choice in surgical treatment of giant benign intrathoracic tumors. Treatment policy is determined individually on the basis of the results of CT, functional and intraoperative findings. PMID- 16353022 TI - [Prophylaxis of insufficiency of gastrointestinal anastomoses]. AB - Experimental and clinical results of use of fibrin glue for strengthening "high risk" sutures of anastomoses in elective surgery of the esophagus, stomach and urgent abdominal surgery are presented. In experimental studies this glue permitted to increase mechanical strength and containment of anastomosis, to accelerate microbial elimination, activated reparative processes. In clinical practice fibrin glue permitted to avoid suture insufficiency of gastrointestinal anastomoses in the most unfavorable conditions (peritonitis, acute intestinal obstruction, oncological process, etc). It is concluded that fibrin glue is the high effective protective agent for gastrointestinal anastomoses. PMID- 16353023 TI - [Choice of surgical strategy in giant gastric and duodenal ulcers]. AB - The results of surgical treatment of 361 patients with giant ulcers of the stomach (54.5%) and duodenum (41.3%) were analyzed. Emergency (within 1 day) surgery was performed in 179 (49.6%) patients, urgent (within 3 days) surgery -- in 158 (43.8%). Active surgical policy decreases lethality and complications rate. Resection of the stomach is the surgery of choice. Palliative surgery aims at stabilization of the patient's condition in critical situation. PMID- 16353024 TI - [Surgical strategies in perforated of gastroduodenal ulcers combined with penetration and stenosis]. AB - One hundred and twenty-six patients aged 14 to 88 years underwent surgery concerning perforative ulcer os stomach and duodenum. Forty-seven patients had symptoms of stenosis including 23 patients with penetration. It is demonstrated that vagotomy, incision of ulcer with duodenoplasty or pyloro-duodenoplasty permit to treat successfully perforative ulcers and to achieve good functional results in long-term period. PMID- 16353025 TI - [Comparative evaluation of different methods of esophagogastroplasty]. AB - The experience in 102 cases of esophagogastroplasty is analyzed, different methods of formation of gastric transplant both in tumors and cicatricial stenosis of the esophagus and stomach are described. Advantages and disadvantages of 57 intrathoracic and 45 cervical esophago-gastric anastomoses with different methods of gastric tube formation are also described in detail. Overall lethality was 5.9%. PMID- 16353026 TI - [Reconstructive-reparative surgeries on the esophagus in burn cicatricial strictures]. AB - Esophagoplasty was performed in 196 patients. Combined disease of the esophagus and stomach was in 71 patients. Esophagogastroplasty was performed in 122 patients, including 104 with gastric tube, 12 -- with whole stomach, 6 -- by means of lengthening of whole stomach. Insufficiency of cervical anastomosis was revealed in 17.2% cases. Long-term results (from 6 months to 9 years) were studied in 77.9% patients. Lethality was 4.2%. Esophagocoloplasty was performed in 74 patients. Insufficiency of cervical anastomosis was seen in 14.5% cases. Long-term results (from 5 months to 9 years) are available in 87.8% patients. Lethality was 4.1%. PMID- 16353027 TI - [Methods of surgical and pharmacotherapeutic influence on the celiac plexus in abdominal surgery]. AB - Literature and own experience in surgical, surgical-pharmacological and contact electro-pharmacological influence on the celiac plexus for correction of dysfunctions in the upper abdominal diseases are analyzed. Original surgical procedures on sympathetic nerves in celiac neuro-ischemic syndrome, chronic duodenal obstruction, duodenal ulcer, and also methods of catheterization and electro-pharmacological impact on celiac plexus in celiac ganglioneuritis and surgical trauma of the stomach are described. PMID- 16353028 TI - [Extrawound programmed vulnosynthesis in the treatment of festering wounds]. AB - The treatment of 200 patients with granulating wound of soft tissues was analyzed. A conventional method of treatment was used in 90 patients. In the study group (110 patients) different types of adaptive-reposition devices and the method of extrawound programmed vulnosynthesis were used. This method demonstrated technical advantages compared with standard methods of treatment of festering wounds that permitted to improve the results of complex treatment. PMID- 16353029 TI - [Bioamine regulation mechanism periodization of teeth development]. AB - Participation of biogenic amines, in cells of dental pulp, in processes of tooth development regulation was studied. The connection was found between contents and inactivation of biogenic amines in developing tooth, as well as in bioamine producing cells and nervous fibres, which had natural differences depending on periods of germ development. PMID- 16353030 TI - [Changes in gum in cases of arterial hypertension combination with periodontitis]. AB - Microlymphohemocirculatory bed and leucocytogram of gingival tissue were studied by the light microscopy in patients with chronic periodontitis having normal and high arterial blood pressure. In most cases of arterial hypertension the gingival mucous was characterized by widening of lymphatic vessels and interstitial spaces. In cases of arterial hypertension combination with inflammatory reaction the tendency for widening of lymphatic vessels and interstitial spaces persisted compared with cases of normal blood pressure. It testifies to high probability of lymphogenic generalization of inflammation. Besides, in cases of inflammatory gingival pathology in arterial hypertension the absolute neutrophil number was significantly higher showing for more acute inflammatory process and greater volume of tissue involvement. PMID- 16353031 TI - [Experience in medical ozone use for root canal treatment]. AB - The results of clinical and laboratory assessment of effectiveness of PSR diagnosis are presented. The high efficacy of ozone therapy is revealed. The findings demonstrate the reduction of number in the micro-organisms in root canal: Actinobacillus actinomycetemcomitans from 31.25% to 10.21%; Bacteriodes forsythus from 68.75% to 15.50%; Treponema denticola from 37.5% to 11.4%; Porphyromonas gingivalis from 56.25% to 45%; Prevotella intermedia from 16% to 0%. PMID- 16353032 TI - [Clinical results of potassium nitrate use in polycarboxylate cement for biological treatment of reversible pulpitis]. AB - To determine effectiveness of indirect pulp capping with 5% potassium nitrate in polycarboxylate cement for biological treatment of reversible pulpitis, a clinical study of 232 teeth with active caries lesions from 205 healthy patients aged from 16 to 40 years was performed. They were divided into 2 groups: the study group--140 teeth of 127 persons with 5% potassium nitrate in polycarboxylate cement (Carboxy Adhesor); control group--92 teeth of 78 persons with calcium-hydroxide cement (Dycal). Short-term clinical results showed better desensitizing effect and faster recovery to normal values of electric vitality in teeth treated by potassium nitrate in comparison with Dycal lining (p<0.001). Long-term follow-up (up to 2 years) showed good functional conditions, preserved vitality and x-ray status of the treated teeth. PMID- 16353033 TI - [Kuttner's inflammatory tumor (sialadenosis of submandibular salivary glands). Case report]. AB - A clinical case of a observation over the patient with Kuttner's inflammatory tumor is described. Quick and complete curative effect was achieved after the use of one course of hyperbaric oxygenotherapy. PMID- 16353034 TI - [Regional metastasizing of maxillofacial region malignant tumors according to the findings of in-unit-form removed tissues study following cervical lymphadenectomy operations]. AB - Changes in regional lymphatic nodes according to N-sign and results of histologic study of these lymphatic nodes in unitized removed tissues following 264 performed operations of cervical lymphadenectomy were studies depending on location and occurrence of primary maxillofacial tumours. PMID- 16353035 TI - [Optimal reconstruction of destructed teeth by pin constructions]. AB - 327 patients with dentures supported by 318 pulpless teeth reinforced by pin constructions were under observation. Main types of complications after pin use were identified. By math simulation the system "covering construction-artificial stump-root pin-root of the tooth" was studied for teeth 11, 13, 15, 16, 41, 43, 45, 46 in two projections--vestibulo-oral and mesio-distal. Margins of strength properties of teeth armed by special pin constructions (made as individual casting) and artificial stump with standard pins made of different materials were determined. Optimal parameters of height and width of off-root part and intra root pin sizes in different teeth groups in conformity with their individual anatomic peculiarities were established. PMID- 16353036 TI - [Early loading of the dental implants installed within 6 months after tooth extraction]. AB - In the period of 6 months after tooth 237 dental implants were installed. The loading was implemented in 1-4 weeks, 2-3 months and also 4-8 months after the operation. A significant correlation of the results of the early charge on the implants with the type of prosthetic construction was found out. The artificial crowns which splint dental implants and natural teeth proved to be the most effective. By such prosthetic treatment the successful outcomes were achieved the more often the earlier the charge begins. The results of the examination confirm the expediency of early dental implantation and early loading on implants. PMID- 16353037 TI - [Correction mechanisms in Angle class II subclass I malocclusion in persons of different age]. AB - The study of 6 groups of patients with an Angle Class II, Division I malocclusions different in age and a method of treatment was made. Mechanisms of correction of an Angle Class II, Division I malocclusion which are typical for each group were determined. The knowledge of mechanisms of correction allows to plan stages of treatment from early primary up to a permanent dentition. PMID- 16353038 TI - [Caries prevention in children during orthodontic treatment]. AB - The study allowed establishing imbalance of immune system during orthodontic appliance treatment. It follows in 1, 5 month after fixation of appliance and in the final period of treatment (15 months after fixation of breakers). These can be considered "critical" and it is necessary to make immune-correctional therapy and to prescribe measures of caries prevention. PMID- 16353039 TI - [Changes of functional-diagnostic indices during occlusive contacts reestablishment in the process of orthodontic treatment of patients with incisal disocclusion]. AB - During orthodontic treatment of 42 patients with incisal disocclusions of different severity stages it was determined that during the period of occlusal contacts reestablishment a considerable increase of regional blood flow, a decrease of jaw bone density and an increase of masticatory muscles functional activity occurred. These changes were more pronounced in cases of deep incisal disocclusion than in cases with other types of incisal disocclusion. PMID- 16353040 TI - [Treatment of chronic heart failure. The time of statins?]. PMID- 16353041 TI - [Indirect comparison of changes of parameters of hemostasis during short-term use of ticlopidine and clopidogrel in patients with non-ST elevation acute coronary syndrome]. AB - Effects of thienopyridines ticlopidine (TIC) and clopidogrel (CL) on hemostasis in patients (pts) with non-ST-elevation acute coronary syndromes (NSTEACS) have not been compared. AIM: To compare changes of some markers of coagulation and platelet activation during short term use of TIC and CL in pts with NSTEACS. METHODS: Aspirin treated pts with NSTEACS (<48 hours from pain onset, Braunwald class IIIb) were included into 2 consecutive studies: 37 pts receiving unfractionated heparin (UFH) were randomized to open TIC (n=19, 500 mg BID for 2 days and 250 mg BID for subsequent 5 days) or no TIC (n=18); 19 pts receiving enoxaparin were randomized to CL (n=10, 300 mg on day 1 and 75 mg/day for subsequent 6 days) or no CL (n=9). At baseline, on days 1, 3, 7 and 14 (7 days after thienopyridines discontinuation) we measured ADP-induced and spontaneous platelet aggregation (PA), levels of prothrombin fragment 1+2 (F1+2), thrombin antithrombin complex (TAT), von Willebrand factor (vWF), fibrinogen, tissue type plasminogen activator antigen (tPA), plasminogen activator inhibitor activity (PAI) and D-dimer (Dd), and counted platelet number. RESULTS: Maximal suppression of PA was obtained on 7-th and 3-rd days in TIC and CL groups, respectively. Compared with their controls TIC treated pts in 7 days after TIC discontinuation had lower levels of TAT (3.61 and 2.77 ng/ml, respectively, r<0.05) and fibrinogen (3.84 and 3.16 g/l, respectively, r<0.05). There were no significant differences between intervention and control groups in these parameters in study with CL. Level of vWF in TIC treated pts was lower than in controls on days 3 (163 and 186%, respectively, r<0.05) and 14 (144 and 173%, respectively, p<0.01). In CL treated pts vWF level was lower relative to controls on days 3 and 7 (152 and 185%, r<0.05, 141 and 166%, r<0.05, respectively). tPA levels in study with TIC did not differ between intervention and control groups. tPA in CL treated pts exceeded its level in controls on days 3, 7, and 14 (25.7 and 20.2 ng/ml, 26.5 and 12.9 ng/ml, 24.6 and 15.7 ng/ml, respectively). On the same days level of Dd in pts receiving CL was significantly higher than in control group (969 and 702 ng/ml, 970 and 575 ng/ml, 806 and 484 ng/ml on days 3, 7 and 14, respectively). Activity of PAI in TIC group was higher than in controls on day 7 (13.6 and 8.2 U/l, r<0.05), and at this moment level of Dd was lower in TIC treated patient (770 and 515 ng/ml in control and TIC groups, respectively, r<0.05). CL and control groups had similar PAI activity. Mean platelet volume rose relative to initial level and to control group only in CL treated patients (9.0 and 8.4 fl, 9.6 and 8.4 fl, 9.4 and 8.5 fl in CL and control groups on days 0, 7 and 14, respectively; r<0.05 for comparison between groups on days 7 and 14). CONCLUSION: In pts with NSTEACS both thienopyridines attenuated acute phase elevation of vWF. The use of TIC in UFH treated pts was associated with indirect signs of decreased thrombin activity and some inhibition of fibrinolysis while the use of CL in enoxaparin treated pts was associated with signs of activation of fibrinolysis. PMID- 16353042 TI - [Associations of hemostasis factors genes with early development of ischemic heart disease and manifestation of myocardial infarction in young age]. AB - AIM: To study polymorphisms of genes of factors of the system of hemostasis in young patients with ischemic heart disease (IHD). MATERIAL: Two groups of patients participated in the study: patients with first manifestation of IHD at the age < or = 50 years (men) or < or = 55 years (women) (n=158), and patients with first IHD manifestation at the age > or = 70 years (n=92). METHODS: We studied polymorphic markers of genes encoding clotting factors V (F5) and VII (F7), subunit IIIa of platelet integrin (ITGB3), beta-chain of fibrinogen (FGB) and tissue plasminogen activator type 1 (PLANH1). RESULTS: After separation of a subgroup of patients with MI without preceding angina we revealed significant differences in distribution of frequencies of genotypes of polymorphic marker C( 426)T of factor V gene: genotype TT was significantly more frequent in young (14.9%) than in old (2%) patients (p=0.008). Multifactorial logistic regression revealed independent association of early IHD with smoking (OR 6.112 [2.567 14.552]; p<0.001) and presence of genotype TT of C(-426)T polymorphic marker of F5 gene (OR=9.410 [1.074-82.459]; p=0.043). CONCLUSION: Thus we obtained data on the presence of independent association between IHD risk and manifestation of MI in young age with genotype TT of polymorphic marker C(-426)T of F5 gene as well as with traditional risk factors of IHD. PMID- 16353043 TI - [Interrelationship between functional state of the right ventricle and severity of heart failure in patients with ischemic heart disease and depressed left ventricular function]. AB - Dobutamine Doppler echocardiography was carried out in 56 patients (n=56) with ischemic heart disease and depressed left ventricular function (left ventricular ejection fraction <40%) and chronic heart failure. Clinical signs of heart failure were moderate (NYHA class I-II) in 34 and severe (NYHA class III-IV) in 22 patients. Patients with moderate and severe clinical heart failure had similar degree of left ventricular myocardium impairment however those with severe symptoms had more pronounced right ventricular (RV) dysfunction (greater suppression of global and local RV contractility, greater percentage of irreversibly dysfunctional RV myocardium, lower RV contractile response to dobutamine infusion, more pronounced disturbances of RV diastolic filling). Dependence of RV pump function on pulmonary artery pressure was more evident in patients with severe clinical heart failure and marked dysfunction of RV myocardium than in patients with moderate symptoms and moderate RV myocardial dysfunction. PMID- 16353044 TI - [Changes of elastic properties of magistral arteries in patients with ischemic heart disease during treatment with atorvastatin and simvastatin]. AB - AIM: To elucidate effect of atorvastatin and simvastatin on elastic properties and reactivity of large arteries in patients with ischemic heart disease. MATERIAL: Patients (n=38) with ischemic heart disease receiving atorvastatin (10 mg/day, n=20) and simvastatin (20 mg/day, n=18). METHODS: Elasticity of arteries was assessed by pulse wave propagation velocity measured by Coloson computer system. Test with reactive hyperemia was used for assessment of large muscular type vessels reactivity in response to blood flow augmentation was assessed by. RESULTS: Treatment with statins for 6 months was associated with positive changes of elasticity of arteries of elastic type (-10.05% and -4.66% in atorvastatin and simvastatin treated patients, respectively) and endothelium dependent reactivity of arteries of muscular type (-16.01%, p<0.001 and -11.1% p<0.001 compared with initial values in atorvastatin and simvastatin treated patients, respectively). CONCLUSION: Therapy with statins positively affects parameters of elasticity and reactivity of large arteries. PMID- 16353045 TI - [Assessment of antihypertensive efficacy of moexipril in metabolic syndrome]. PMID- 16353046 TI - [Comparative efficacy of fixed dose combinations of enalapril with hydrochlorothiazide and captopril with hydrochlorothiazide in patients with high risk hypertension]. AB - Clinical effectiveness and tolerability of o.d. use of fixed dose combinations of enalapril (10 mg) with hydrochlorothiazide (25 mg) (Enap H) and captopril (50 mg) with hydrochlorothiazide (25 mg) (Capozide) were compared in a randomized study on 60 patients with I-II degree high and very high risk hypertension. Study duration was 6 months, number of patients in each of parallel groups -- 30. Antihypertensive activity, ability to improve arterial elasticity and T/P parameter, cost/efficacy index of enalapril (10 mg) plus hydrochlorothiazide (25 mg) combination was found to be superior to those of captopril (50 mg) plus hydrochlorothiazide (25 mg) combination. PMID- 16353047 TI - [Remote catamnesis of paroxysmal supraventricular arrhythmias which debuted in childhood]. AB - This article deals with the 23-years assessment of natural history of 57 patients with paroxysmal supraventricular tachycardia debuted in childhood. Variants of clinical course of disease are described: cardiac, syncopal, abdominal, asymptomatic. In absence of treatment majority of paroxysmal tachycardias were characterized by wavy change of their clinical manifestations with periods of pronouncedly increased and decreased activity lasting from 3 to 5 years. Time interval between attacks was important for prognosis. Severity of disease was determined by relationship of three factors: frequency, duration of attacks and ability of a tachycardia to cause disturbances of central hemodynamics. Tachycardia with asynchronous AB conduction and heart rate above 250/min in one year old and above 220/min in older children was associated with increased risk of development of acute heart failure during an attack. Risk of heart failure was not related directly to frequency of attacks but arose when duration of tachycardia with critical heart rate exceeded 8 hours. PMID- 16353048 TI - [Correction of anxiety depressive disorders with tianeptine in patients with heart failure in early period after myocardial infarction]. AB - Aim of the study was clarification of the role of psychological characteristics of personality of patients, relationship between parameters of quality of life and clinical characteristics of the disease and their effect on peculiarities of the course of curative-rehabilitational process as well as clarification of spectrum of clinical activity and safety of tianeptine in anxiety-depressive states in patients with chronic heart failure in early post infarction period. Of 80 patients with anxiety-depressive disorders studied on days 15-30 after onset of disease, 30 received tianeptine (37.5 mg/day) for 3-6 months and 50 comprised control group. Significant reduction of symptoms of anxiety and depression and of cognitive-mnestic functions which occurred in tianeptine treated patients was accompanied with diminished severity of symptoms of heart failure and coronary insufficiency, lowering of index of local myocardial contractility and growth of ejection fraction. All of this resulted in improvement of quality of life of postinfarction patients, shortening of terms of rehabilitational period and allowed the patients to adapt better to surrounding reality. PMID- 16353049 TI - [Clinical value of episodes of acute lowering of heart rate variability detected during 24-hour ECG monitoring in patients with ischemic heart disease]. PMID- 16353050 TI - [Effect of therapy with antibiotics on lipid metabolism and antioxidant reserve of patients with ischemic heart disease during Chlamydia pneumoniae infection]. AB - Effect of therapy with azithromycin and doxycycline on lipid metabolism, processes of lipid peroxidation and state of antioxidant defense was studied in patients with ischemic heart disease with elevated titer of antibodies to Chlamydia Pneumonia. Therapy with azithromycin (500 mg/day for 2 months) was associated with lowering of antibody titer, moderate improvement of lipid spectrum, marked decrease of activity of lipid peroxidation and augmentation of blood plasma antioxidant reserve. There were no such changes in a group of doxycycline treated patients. This difference can be attributed to more pronounced antimicrobial activity of azithromycin against Chlamydophila Pneumonia and its intrinsic antioxidant properties. PMID- 16353051 TI - [Molecular mechanisms of dysfunction of vascular endothelium]. PMID- 16353052 TI - [Marfan syndrome in a practice of a cardiologist: diagnosis, treatment, possibility of pregnancy and delivery]. PMID- 16353053 TI - [Ischemic heart disease combined with hypertension: peculiarities of course and selection of therapy]. PMID- 16353054 TI - [Novel possibilities of antithrombotic therapy in patients with chronic heart failure]. AB - Modern approaches to prevention of venous thromboembolic complications in patients with chronic heart failure are analyzed in this review which contains results of large studies of low molecular weight heparins. In MEDENOX trial the use of enoxaparin in medical patients was associated with 63% reduction of risk of thrombosis. The authors own experience showed that 2 weeks of therapy with enoxaparin in patients with chronic stage IIB-III heart failure caused significant lowering of soluble fibrin-monomer complexes, fibrinogen, and index of turbo-dynamic potential. These changes evidenced for decreased intravascular blood coagulation. Thus enoxaparin can be effectively used for prevention of thrombosis and thromboembolism in patients with chronic heart failure. Novel antithrombotic agents fondaparinux, idraparinux, ximelagatran, recombinant thrombomodulin are perspective medications for prevention of venous thromboses and embolism in medical patients. PMID- 16353055 TI - [Hypertension combined with atherosclerotic lesions in carotid arteries: should angiotensin converting enzyme inhibitors be prescribed?]. PMID- 16353056 TI - [Surgical treatment of a patient with aneurysm of ascending aorta and involvement of three cardiac valves due to Marfan syndrome]. PMID- 16353057 TI - [Clinico-epidemiological program of the study of depression in cardiological practice in patients with hypertension and ischemic heart disease: first results of a multicenter study]. PMID- 16353058 TI - [Structural-functional changes of myocardium and hemodynamic disturbances in patients with metabolic syndrome: contribution of arterial hypertension in formation of total coronary risk]. AB - AIM: To assess in the aggregate hemodynamic peculiarities and changes of the myocardium as well as contribution of hypertension in formation of coronary risk in metabolic syndrome (MS). MATERIAL AND METHODS: Patients (n=290) with hypertension of I-II degree and duration > or = 5 years were subjected to laboratory (parameters of lipid spectrum, glucose and insulin levels) and instrumental (24-hour blood pressure monitoring, echocardiography) examination. Criteria of MS were fasting insulin > 18 mcU/ml and/or glucose/insulin ratio < 6; blood pressure (BP) > or = 140/90 mm Hg; triglycerides > or = 200 mg/dl and/or high density lipoprotein cholesterol < 39 mg/dl; body mass index > 25 kg/m(2) with waist/hip circumference ratio > or = 0.95 (for men) or > or = 0.80 (for women); impaired glucose tolerance according to WHO criteria. PROCAM model was used for calculation of total coronary risk. RESULTS: 37% of patients with hypertension had all components of MS. Hypertension took third place among contributors to formation of total coronary risk with input of 20%. Diastolic BP positively correlated with triglycerides, total cholesterol, index of insulin resistance and parameters of abdominal obesity. Mean 24 hour systolic and pulse BP in patients with MS were significantly higher than in a group of patients with hypertension without metabolic disturbances. Patients with MS had thicker left ventricular posterior wall and interventricular septum what was associated with increased end-systolic and end-diastolic dimensions as well as myocardial mass of the left ventricle. CONCLUSION: In patients with MS 24 hour BP profile is impaired at the account of high pulse BP and lack of nocturnal lowering of systolic and diastolic BP and is associated with concentric left ventricular hypertrophy. PMID- 16353059 TI - [Antihypertensive activity and effect on parameters of heart rate variability of beta-blockers carvedilol and metoprolol]. PMID- 16353060 TI - [Comparative efficacy of fixed dose combinations of perindopril with indapamide and captopril with hydrochlorothiazide in patients with high risk hypertension]. AB - Clinical effectiveness and tolerability of o.d. use of fixed dose combinations of perindopril (4 mg) with indapamide (1.25 mg) (Noliprel forte) and captopril (50 mg) with hydrochlorothiazide (25 mg) (Capozide) were compared in a randomized study on 40 patients with I-II degree high and very high risk hypertension. Study duration was 6 months, number of patients in each of parallel groups -- 20. Antihypertensive activity, ability to decrease left ventricular hypertrophy, to improve arterial elasticity and T/P parameter of perindopril (4 mg) -- indapamide (1.25 mg) combination was found to be superior to those of captopril (50 mg) -- hydrochlorothiazide (25 mg) combination. PMID- 16353061 TI - [Special characteristics of pathogenesis of hypertension in personnel of radiation dangerous industry]. PMID- 16353063 TI - [Possibility of coronary angiography without hospitalization]. AB - Coronary angiography is the necessary investigation in the diagnostic complex of patients with ischemic heart disease (IHD). The use of trans-radial access makes it possible to do the angiography without hospitalization. The algorithm of coronary angiography in day-time clinic was proposed. Non-hospital angiography in 15 patients was successful. It is the first time this method has been used in this country. PMID- 16353062 TI - [Physical activity and atherosclerosis: effect of dynamic activity of various intensity on parameters of lipid-transport system and carbohydrate metabolism in patients with coronary heart disease and type 2 diabetes]. AB - Effect of single bout of dynamic physical exercise on parameters of lipid transport system and carbohydrate metabolism and hormones (insulin, cortisol) in the blood was studied in patients with coronary heart disease with class I-III angina and type 2 diabetes. Intensity of exercise was limited by severity of stable effort angina and was > 95, 80 and 70% of predicted maximum in patients with class I (n=10), II (n=12) and III (n=14) angina, respectively. High intensity exercise provoked development of atherogenic dyslipidemia: elevation of levels of total cholesterol, low density lipoprotein cholesterol, triglycerides, apolipoprotein B and apolipoprotein B/A1 ratio, and lowering of levels of high density lipoprotein cholesterol and apolipoprotein A1. Patients with diabetes responded to high intensity exercise by elevation of blood glucose and insulin levels and lowering of sensitivity of tissues of the periphery to insulin (glucose/insulin ratio). On the contrary exercise of moderate intensity did not affect negatively metabolism of blood lipids and carbohydrates. Six months course of physical training in patients with diabetes (n=10) corrected exogenous atherogenic dyslipidemia and derangements of carbohydrate metabolism, which developed after acute dynamic effort of high intensity. PMID- 16353064 TI - [Selection of therapy with nitrates in patients with stable effort angina: results of comparative study of common isosorbide dinitrate and long acting preparation of isosorbide-5-mononitrate]. AB - AIM: To assess efficacy and tolerability of a novel drug form of isosorbide-5 mononitrate in patients with ischemic heart disease and stable effort angina as compared with common isosorbide dinitrate pills. MATERIAL AND METHODS: Patients with stable class II-III effort angina (n=30) were included into a randomized crossover study in which they received isosorbide dinitrate (nitrosorbide, 10-20 mg t.i.d.) and long acting isosorbide-5-mononitrate (ephox-long, 50-100 mg o.d.) for 3 weeks each. Efficacy of treatment was assessed by clinical data and treadmill exercise tests. Questionnaires were used for registration of frequency and intensity of attacks of headache. RESULTS: The use of both isosorbide dinitrate and 5-mononitrate was associated with significant improvements of exercise tolerance however effect of mononitrate lasted longer. Nitroglycerine requirement diminished during first week of use of both drugs and remained on this level by the end of 3-rd week of treatment with mononitrate but substantially rose by the end of treatment with dinitrate. Number of attacks of headache increased during first week of treatment with both drugs, became even higher by the end of use of dinitrate and decreased by the end of use of mononitrate. CONCLUSION: Long acting form of isosorbide-5-mononitrate ephox-long taken once daily provides sufficient antianginal effect throughout a day and is better tolerated than nitrosorbide preparation of isosorbide dinitrate with moderately prolonged activity. PMID- 16353065 TI - [Disturbances of cardiac rhythm and metabolic control in patients with type-2 diabetes]. AB - Twenty four hour ECG monitoring, registration of ventricular late potentials, and measurement of glycated hemoglobin (HbA1c) were included into examination of 142 cardiological patients (101 with and 41 without type 2 diabetes). Groups of patients with and without diabetes had similar age and severity of cardiac pathology. Patients with diabetes had more prognostically and hemodynamically unfavorable arrhythmias -- paroxysmal and permanent atrial fibrillation, high grade ventricular extrasystoles (HGVE) and their combinations. Atrial fibrillation and HGVE were more often observed in diabetic patients with HbA1c <7 and >8.5%, respectively. Date of registration of ventricular late potentials in patients with diabetes mellitus (QRSTt >110 ms, LAS40 >37 ms, and RMS <23 mcV) possess high informative power not only in prognosis of HGVE but also evidences for bad control of glycemia. PMID- 16353066 TI - [Comparative assessment of the use of skeletonized and un-skeletonized internal thoracic artery for myocardial revascularization]. AB - BACKGROUND: Skeleting of internal thoracic artery widens possibilities of a surgeon during mammary-coronary bypass grafting but this procedure is associated with augmented risk of damage of internal thoracic artery (ITA). MATERIAL AND METHODS: ITA was used for creation of anastomosis with anterior interventricular artery in 151 patients. It was harvested on pedicle in 82 (group 1) and subjected to skeleting in 69 (group 2) patients. All operations were performed with cardiopulmonary bypass. In group 2 ITA conduits were more frequently used for grafting distal anterior interventricular artery. RESULTS: There were 3 and 0 deaths in groups 1 and 2, respectively. Rates of nonfatal postoperative complications were similar in both groups. Spasms of ITA grafts were more frequent in group 1. CONCLUSION: Skeleting of ITA for mammary coronary bypass grafting compared with its harvesting on pedicle was not associated with increased risk of complications or elevated mortality in early postoperative period. PMID- 16353067 TI - [Heart rate and QT-interval dispersion with consideration of left ventricular geometry in a population study of men and women aged 35-59 years]. AB - Interrelationships between heart rate (HR), parameters of QT interval dispersion on resting ECG and four variants of echocardiographically defined left ventricular geometry were elucidated in 723 participants of a population study of inhabitants of Tallinn aged 35-59 years. Examination included anthropometry, blood pressure (BP) measurement, registration of ECG and echocardiography. None of normotensive examinees had concentric left ventricular hypertrophy. Among men with normal BP eccentric left ventricular hypertrophy was very rare (0.88%) and contrary to normal left ventricular geometry was associated with highest values of HR and QTD, QTc, QTDc. Among normotensive women eccentric left ventricular hypertrophy was found in 35.3% of cases and was associated with lowest values of HR while parameters of QT interval dispersion were similar to those observed in other variants of left ventricular geometry. Both men and women with hypertension and left ventricular hypertrophy had significantly lower HR and higher values of QT interval dispersion compared with hypertensives with normal left ventricular geometry. In men HR was positively related to age, systolic and diastolic BP and left ventricular myocardial mass index. In women relationship between HR, BP and age was very weak. Relation of HR to left ventricular myocardial mass index was more noticeable although turned out to be inverse. PMID- 16353068 TI - [Myocardial revascularization in patients with type 2 diabetes]. PMID- 16353069 TI - [Stem cells and regeneration of human myocardium]. AB - A concept of impossibility of appearance of novel cardiomyocytes in the heart of adult men in exchange for those lost due to cardiovascular diseases had dominated medicine and biology for many long decades. However ability of human myocardium to regenerate was demonstrated during recent years in multiple studies. This dictated necessity to reconsider previously generally accepted concept. At present researchers and practicing physicians actively discuss possibility of the use of transplantation of bone marrow stem cells, proper cardiac stem cells, skeletal muscle myoblasts or precursors of endothelial cells in patients with myocardial infarction and heart failure in order to restore normal cardiac structure and function. Another potential method of restoration of the myocardium in patients with cardiovascular diseases is the use of cytokines which stimulate migration of stem cells into myocardium and their differentiation into cardiomyocytes. PMID- 16353070 TI - [Transesophageal and transthoracic ultrasound study of coronary arteries in diagnosis of atherosclerosis and assessment of coronary reserve]. AB - The lecture deals with methodological aspects and diagnostic potential of the use of novel ultrasound technologies -- transesophageal and transthoracic doppler ultrasonography of coronary arteries for noninvasive assessment of coronary blood flow and coronary reserve. It contains detailed description of technique of visualization of coronary arteries from transesophageal and from transthoracic approaches and interpretation of dopplero-graphic parameters of laminar and turbulent coronary flows. Diagnostic role of the use of contrast echo imaging of coronary arteries is also discussed and dopplero-graphic criteria of occlusion and hemodynamically significant coronary artery stenosis as well as characteristics of normal and lowered coronary reserve are presented. PMID- 16353071 TI - [Electrical instability of the myocardium in patients with aortic stenosis]. AB - This review of the problems of prognostication of electrical instability of the myocardium in patients with aortic stenosis is based on recommendations of European Society of Cardiology on prevention of sudden death in patients with aortic stenosis with special emphasis on potential of noninvasive assessment of myocardial electric instability (signal averaged ECG, heart rate variability, analysis of variability of QT-interval, dipole electrocardiotopography). PMID- 16353072 TI - [Cardiovascular involvement in rheumatic diseases]. AB - Cardiovascular system involvement with early development of atherosclerosis is characteristic for rheumatic diseases. Among causes of death in various rheumatic diseases cardiovascular pathology also prevails. This paper contains a review of most important studies of impairment of the heart, arterial and venous parts of cardiovascular system in patients with diffuse diseases of connective tissue, rheumatoid arthritis and systemic vasculitides. The role of immune mechanisms, endothelial dysfunction, dyslipidemia in pathogenesis of cardiovascular disturbances with development of myocardial and vascular remodeling in rheumatic diseases is also discussed. Major risk factors of cardiovascular pathology in rheumatic patients are presented. Treatment of a cardiovascular pathology in these patients presumes the use of angiotensin converting enzyme inhibitors, aldosterone antagonists and statins. PMID- 16353073 TI - [Prevalence and risk factors of atherosclerosis in patients with systemic lupus erythematosus]. PMID- 16353074 TI - [Angiotensin converting enzyme inhibitors in prevention and treatment of vascular complications of diabetes]. PMID- 16353075 TI - [Arterial stiffness: parameters, methods of assessment and methodological difficulties]. AB - Arterial stiffness has been shown to be an independent marker of morbidity and cardiovascular mortality in multiple prospective studies of various populations. The purpose of this review is to present different parameters of arterial stiffness (compliance; distensibility; elastic modulus; pulse wave velocity; wave reflection), and to discuss existing difficulties of their application. Means of evaluation of various types of arterial stiffness and principles of their use are also described. Arterial stiffness is not a static but dynamic characteristic, thus prior to its evaluation it is necessary to standardize both the state of a patient and methodology of measurements. Diagnostic value of noninvasive methods of assessment of arterial stiffness is similar to that of invasive techniques, therefore stress can be made on less traumatic investigation of vascular properties. PMID- 16353076 TI - Posterior mediastinal tumors: outcome of surgery. AB - BACKGROUND AND PURPOSE: The incidence of posterior mediastinal tumors relative to all tumors of the mediastinum is 23% to 30%. The posterior mediastinum is a potential space along each side of the vertebral column and adjacent proximal portion of the ribs. Primary tumors of posterior mediastinum are usually neurogenic. The aim of this study was to evaluate different surgical approaches used for the resection of posterior mediastinal tumors, and to assess morbidity, mortality and patients' survival. PATIENTS AND METHODS: Between January 2001 and January 2004, 30 patients with posterior mediastinal tumors were included. CT scan of the chest and CT guided biopsy were done for all patients; whereas MRI was done for suspected intraspinal extension. Posterolateral thoracotomy was the approach used in most of the patients. The Akwari approach was used in most of the patients with Dumbbell tumors. RESULTS: Neurogenic tumors constituted 67% of cases, being neuroblastoma in 60%. The non neurogenic tumors included a heterogenous group of rare tumors (n=10). Dumbbell tumors were found in 10 patients. Neuroblastoma was the commonest tumor to cause intraspinal extension (40%). Wide local excision was done in 13 patients; whereas extended resection was done in the remaining 17 patients. The mean intra-operative blood loss was 800cc and the mean hospital stay was 12 days. The size of the resected tumor ranged from 3 X 4 cm to 30 X 22 cm, 80% of tumors were malignant. Morbidity in relation to the procedures developed in 8 patients (atelectasis, meningitis, paraplegia, Horner syndrome and mild wound sepsis in 4, 1, 1, 1 and 1 of the patients; respectively). One postoperative mortality, due to meningitis was recorded. The overall survival by the end of three years was 87.7% with a mean survival of 30.4 months. The overall disease free survival was 55.9% with a mean disease free survival of 26.2 months. CONCLUSIONS: Posterior mediastinal tumors may reach large size before becoming symptomatic. Complete surgical excision (including adjacent invaded organs) mainly by open technique should be the rule for these patients as there is survival benefit. Great care should be taken when dealing with Dumbbell tumors. PMID- 16353077 TI - Prognostic factors of Hodgkin's lymphoma and their impact on response to chemotherapy and survival. AB - OBJECTIVE: The aim of this study was to compare the standard prognostic factors of Hodgkin's lymphoma (HL) in relation to response to first line chemotherapy, disease free survival (DFS) and overall survival (OS). PATIENTS AND METHODS: The study was performed on a group of 100 adult patients diagnosed as HL and who were treated and followed-up in the years 1999 to 2001, in the Medical Oncology Department at National Cancer Institute (NCI), Cairo. The first line chemotherapy was COPP in 40%, ABVD in 35% and COPP/ABV hybrid in 25%. Patients were classified into early stage disease: Stages I, IIA and IIB without poor risk factors, n=43 and advanced stage disease: Stages III, IV and IIB with poor risk factors, n=57 analysis of the prognostic factors for early versus advanced-stage disease was done by univariate and multivariate regression analysis. RESULTS: Complete remission (CR) was attained in 69% of the patients after first line chemotherapy; being 87.8 % and 54.7% for early and advanced disease, respectively, (p=0.0001). The CR rates after different chemotherapy regimens were 81.8%, 90% and 90% for the ABVD, COPP and COPP/ABV hybrid regimens in the early-disease group; respectively; in contrast to the corresponding figures of 54.5%, 50% and 61.5% in the advanced- stage group. The DFS at 4 years, was 94 %, 55% and 54.5% for the patients treated with ABVD, COPP and COPP/ABV hybrid, respectively (p=0.2). The DFS and OS in this series of patients were 61.3% and 53.7%, being 69.8% and 70.7% for the early and 45.1% and 38.9% for the advanced-disease, respectively The OS of the whole group was significantly related to age (p=0.04), sex (p=0.005), early versus advanced disease (p=0.0001) and B symptoms (p=0.0006). CONCLUSIONS: The adequate response and DFS of the early compared to the advanced-stage disease supported the evolving role of risk adapted chemotherapy for HL. The prognostic factors proved to be of significant impact in our series. The results of this study pointed to the need for an improved treatment strategy in this potentially curable disease,especially for the advanced disease. PMID- 16353078 TI - Precursor B-cell lymphoblastic lymphoma (PBLL) in children: pattern of presentation and outcome. AB - PURPOSE AND BACKGROUND: Precursor B-cell lymphoblastic lymphoma (PBLL) is a rare subtype of NHL seen primarily in children or young adults. There are approximately 100 immunophenotyped cases of PBLL; reported in the literature; most as single case reports or very small series. In this report, we describe patterns of presentation, and results of a retrospective study looking at patients with PBLL treated at KFSH and RC between 1993 and 2000. PATIENTS AND METHODS: We present results of a retrospective study looking at patients with PBLL treated at KFSHRC between 1993 and 2000, younger than 14 years of age (cut off age for pediatric department). Six cases of PBLL were lacking evidence of blood and bone marrow involvement. Histologic sections were available for review in all cases. RESULTS: Twenty one patients were treated for lymphoblastic lymphoma, of which six had a precursor Bcell phenotype. There were three boys and the median age at diagnosis was 6 years (range 3-13). In four of the patients the primary involved were oro-nasopharynx or the paranasal sinuses. One patient had a soft tissue mass in the upper thigh while one patient had a solitary bone lesion in the distal tibia. Four of the patients had limited stage disease (2 stage I and stage II), while 2 were stage IV. Both patients with stage IV disease had CNS involvement with blasts in the CSF. Both had paranasal primaries and had bone infiltration involving the base of the skull, with radiological documentation of intracranial extension in one patient. Median LDH level was 542 IU/L (range 463 5000). Five patients were treated according to B-cell NHL type protocols. Because of the specific diagnosis of PBLL, two of these patients were switched to an ALL type protocol following post induction intensification; one died in remission due to encephalitis, while the other remained in CR almost 2 years after diagnosis. A third patient suffered a loco-regional relapse 17 months after completing first line therapy, and was re-treated on an ALL-type protocol, and currently is in remission 25 months following relapse. The fourth patient, who received 9 months of post induction therapy, remains free of disease 7 years following diagnosis. The fifth patient had local and CNS progression on therapy, and died of his disease. The last patient with a solitary bone lesion was misdiagnosed as Ewings' Sarcoma and received treatment for that disease. He suffered an isolated CNS relapse, and is in CR 12 months following the relapse, on an ALL treatment protocol. CONCLUSION: PBLL is a distinct B-cell NHL which involves extralymphatic sites, with particular predisposition for the upper aerodigestive tract. Patients should not be treated on short intensive protocols used for other B-cell NHL but should receive treatment based on ALL protocols like those for treating T-cell LL. PMID- 16353079 TI - De novo nodal diffuse large B-cell lymphoma: identification of biologic prognostic factors. AB - BACKGROUND: Diffuse large B-cell Lymphoma (DLBCL) represents the most frequent type of non-Hodgkin lymphoma (NHL). Although combination chemotherapy has improved the outcome, long-term cure is now possible for approximately 50% of all patients, making the search for parameters identifying patients at high risk particularly needed. The presence of bcl-2 gene rearrangement in de novo DLBCL suggests a possible follicle center cell origin and perhaps a distinct clinical behavior. This study investigated the frequency and prognostic significance of t(14;18) translocation and bcl-2 protein overexpression in a cohort of patients with de novo nodal DLBCL who where uniformly evaluated and treated. MATERIAL AND METHODS: A total of 40 patients with de novo nodal DLBCL treated at National Cancer Institute (NCI), Cairo University were investigated. Formalin? fixed, paraffin-embedded sections were analyzed for: 1) bcl-2 gene rearrangement including major break point region (mbr) and minor cluster region (mcr) by polymerase chain reaction (PCR), and 2) bcl-2 protein expression by immunohistochemistry using Dako 124 clone. Results were correlated with the clinical features and subsequent clinical course. RESULTS: Bcl-2 gene rearrangement was detected in 8 cases (20%), 2 cases at mbr, and 6 cases at mcr. Bcl-2 protein (>10%) was expressed in 24 cases (60%), irrespective of the presence of t(14;18) translocation. The t(14;18), and bcl-2 protein overexpression were more frequently associated with failure to achieve a complete response to therapy (p=0.008, and 0.04, respectively). DLBCL patients with t(14;18), and bcl-2 protein expression had a significantly reduced 5-year disease free survival (p=0.04, and 0.01, respectively). CONCLUSION: The t(14;18) translocation, and bcl-2 protein expression define a group of DLBCL patients with a poor prognosis, and could be used to tailor treatment, and to identify candidates for therapeutic approaches. Geographic differences in t(14;18) may be related to the difference in distribution of bcl-2 breakpoints. PMID- 16353080 TI - Prevalence of anti human herpes virus-6 IgG and its receptor in acute leukemia (membrane cofactor protein: MCP, CD46). AB - BACKGROUND: CD46 is a membrane cofactor protein, which acts as a cofactor for factor I proteolytic cleavage of C3, so it protects the cells expressing it on their surface from autologous complement attack. It has been recently described as a receptor for HHV-6. Also, it has been shown to be highly expressed on malignant cells as compared to normal cells, thus playing a major role by which these cells, either cells of haematological malignancy or cells of other body cancers, can protect themselves against complement attack so they can survive and metastasize. PATIENTS AND METHODS: This study has been done to detect the seroprevalence of HHV-6 among 47 Egyptian adult cases of acute leukemia using the anti-HHV-6 IgG ELISA serological technique. CD46 receptor expression and immunophenotyping technique were performed using FCM. Twenty nine of the cases were ANLL, while 18 were ALL cases. Sixteen age- and sex-matched control cases were also studied for both anti-HHV-6 IgG and CD46 receptor expression. RESULTS: HHV-6 IgG antibodies were encountered in 29 (100%), 14 (77.8%) and 12 (75%) of the ANLL, ALL and the control group, cases, respectively. CD46 expression was encountered in 21 (72.4%) of the ANLL cases and in 10 (55.6%) of the ALL cases. Concordance between HHV- 6 seropositivity and CD46 expression was encountered in 31 cases (29 positive and 2 negative). Disconcordance was encountered in 16 cases with 14 showing HHV-6 IgG seropositivity with no CD46 expression and 2 showing the reverse. CONCLUSION: The lack of significant correlation between CD46 expression and seropositivity would exclude CD46 expression as a cause of contracting HHV-6 infection in leukemic patients. PMID- 16353081 TI - Contribution of nitric oxide and epidermal growth factor receptor in anti metastatic potential of paclitaxel in human liver cancer cell (HebG2). AB - BACKGROUND: Paclitaxel is a general antineoplastic drug used against different types of experimental and human tumors. Several anti-cancer drugs have been shown to stimulate nitric oxide (NO) production, which has been shown to affect many aspects of tumor biology. OBJECTIVE: This study was initiated to determine if paclitaxel stimulates NO production in HebG2 cells, and if so, whether NO interferes with the metastatic potential of HebG2 cells and contributes to paclitaxel cytotoxicity. In addition, we sought to determine the relationship between NO production and the expression of epidermal growth factor receptor (EGFR) and matrix metaloproteinases (MMPs) in HebG2 cells. MATERIALS AND METHODS: The effects of paclitaxel (0.1-1000 nM) on surviving fraction, NO production and the expression of EGFR, MMP-2 and MMP-9 were studied in human liver cancer cells (HebG2). RESULTS: Paclitaxel resulted in a significant dose dependent decrease in the surviving fraction of HebG2 cells. A 62% and 86% decrease in the surviving fraction was attained at 10nM and 100 nM paclitaxel, respectively. Paclitaxel produced a significant increase in NO production, starting from 1 nM. A 64% and 111% increase in NO production was attained after exposure to 10nM and 100 nM of paclitaxel, respectively. In all of the HebG2 cells treated with paclitaxel (1.0 1000 nM) mRNA specific for EGFR, MMP-2 and MMP-9 were undetectable. However, untreated HebG2 cells and those treated with paclitaxel (0.1 nM) expressed mRNA specific for these markers. CONCLUSION: This study suggests that: (1) increased production of NO may contribute to paclitaxel's cytotoxicity against HebG2 cells, (2) paclitaxel may inhibit tumor metastasis via inhibition of the expression of EGFR and MMPs and (3) an inverse correlation exist between NO production and expression of EGFR and MMPs. PMID- 16353082 TI - Survivin expression in colorectal adenocarcinoma using tissue microarray. AB - BACKGROUND: The additional prognostic information closely related to tumor cell biology is essential for the identification of patients with poor prognosis. Survivin, an identified inhibitor of apoptosis, is unique for its expression in human malignancies but not in normal adult cells. This study examined the expression, and potential prognostic value of survivin in colorectal adenocarcinoma (CRC) on tissue microarray (TMA) sections. Analysis of large numbers of tissue samples, improved tissue salvage, cost reduction, ease of interpretation, and significant time saving were realized by using the arrays. MATERIAL AND METHODS: Two-hundred and eighty cases of colorectal adenocarcinoma were arrayed. Immunohistochemical stains of TMA sections were performed for survivin, bcl-2, and p53. Cases were followed up for 5 years. RESULTS: Survivin was detected in 147 of 230 cases (63.9%). No expression of survivin was observed in normal tissues. There was no correlation between survivin immunoreactivity and age, sex, tumor site, tumor size, histopathologic subtype, tumor grade and clinical stage (p >0.05). Prevalence of survivin expression was significantly higher in bcl-2 positive than in bcl-2 negative cases (88.1% versus 42.1%, p <0.0001), but was not associated with p53 (p=0.09). The 5-year disease free survival (DFS) for patients with survivin positive colorectal adenocarcinoma was significantly lower than that for patients with survivin negative tumors (46% versus 68.7%, p=0.001). CONCLUSION: Survivin expression in colorectal adenocarcinoma provides an important prognostic parameter and targeted antagonists of survivin may be beneficial as apoptosis-based therapy for colon cancer. PMID- 16353083 TI - Importance of serum IL-18 and RANTES as markers for breast carcinoma progression. AB - BACKGROUND: Interleukin-18 (IL-18), a cytokine that plays an important role in the T-cell-helper response, acts as an angiogenic factor and a tumor suppressor. RANTES (regulated upon activation normal T-cells expressed and secreted) is a member of the C-C chemokine family with chemoattractant activity for a variety of cell types. High incidence and intensity of RANTES were noted in advanced breast carcinoma. AIM OF THE STUDY: To correlate the levels of RANTES and IL-18 in serum of breast cancer patients with bone or other organ metastasis compared to breast cancer patients without metastasis and healthy controls and to estimate the role of each of them as a prognostic marker for the progression of the disease. PATIENTS AND METHODS: The study was conducted on 60 breast cancer patients (25 cases with no metastasis and 35 cases with metastasis) who were admitted to the outpatient clinic of the NCI, Cairo University during the period from March 2004 to September 2004 and 30 apparently healthy controls who were volunteers at the blood bank of the NCI, Cairo University. RESULTS: Showed that there was a statistically significant difference between the level of IL-18 in breast cancer patients without metastasis and the control group (p<0.05) while there was a highly significant difference between the metastatic group and the control group (p<0.001). There was a significant increase in IL-18 levels between metastatic and non-metastatic cases (p<0.01). RANTES showed a significant increase in breast cancer cases with no metastasis and the control group (p<0.05) and it showed a highly significant increase in metastatic patients compared to controls (p<0.001). There was no significant increase in the level of RANTES in metastatic compared to non-metastatic patients (p>0.05). CONCLUSIONS: IL-18 is an important non invasive marker suspecting metastasis. Even though RANTES levels were higher in cancer patients compared to controls, its role in staging of breast cancer was not clear in this study. PMID- 16353085 TI - Advances in glycoprotein synthesis. AB - The development of chemical and enzymatic methods for the synthesis of homogeneous glycoproteins is a fascinating challenge at the interface between chemistry and biology. Discussed here are the currently available methods for preparation of homogeneous glycoproteins. These methods include (1) glycopeptide ligation; (2) glycoprotein remodeling; and (3) in vivo suppressor tRNA technology. PMID- 16353086 TI - Dendritic supermolecules--towards controllable nanomaterials. AB - Dendritic molecules constitute one of the most exciting areas of modern nanochemistry, largely as a consequence of the unique properties associated with their branched architectures. This article describes how 'dendritic function' can also be achieved using small, synthetically accessible branched building blocks (individual dendrons) which simply self-assemble via non-covalent interactions to generate dendritic nanoscale architectures with novel behaviour. (a) Using non covalent interactions at the focal point of a dendron allows the self-assembly of nanometre-sized supramolecular dendrimers around an appropriate template species. Such systems have potential applications in the controlled encapsulation and release of active ingredients. (b) Employing non-covalent intermolecular dendron dendron interactions can give rise to the hierarchical assembly of nanostructured materials. Such assemblies of dendritic molecules ultimately express their molecular scale information on a macroscopic scale, and therefore have applications in materials science, for example as gels. (c) The multiple surface groups of dendrons are capable of forming multiple interactions with large surfaces, such as those found on biomolecules or in biological systems. Employing multivalent interactions between dendron surfaces and biological molecules opens up the potential application of dendritic systems as medicinal therapies. In summary, dendritic supermolecules offer a potentially cost-effective approach to the future application of dendritic systems to a range of real-world problems. PMID- 16353087 TI - A steep one-step [HS-HS] to [LS-LS] spin transition in a 4,4'-bipyridine linked one-dimensional coordination polymer constructed from a pyrazolato bridged Fe(II) dimer. AB - The variable temperature magnetic susceptibility, X-ray crystallography, and Mossbauer and Raman spectra of a new dinuclear complex-based one-dimensional coordination polymer [[Fe(II)2(NCS)2(mu-bpypz)2](micro-4,4'-bpy)].MeOH demonstrated a steep one-step [HS-HS] to [LS-LS] spin transition. PMID- 16353088 TI - A water-soluble hexa-peri-hexabenzocoronene: synthesis, self-assembly and role as template for porous silica with aligned nanochannels. AB - A water-soluble hexa-peri-hexabenzocoronene was prepared and shown to undergo ordered columnar self-assembly either in water solution or bulk and therefore served as template for the fabrication of porous silica with aligned nanochannels. PMID- 16353089 TI - Screening for crystalline salts via mechanochemistry. AB - Neat grinding and solvent-drop grinding methods are found to be effective screening tools for indicating the potential for crystalline salt formation involving a given acid-base pair, as demonstrated with two model pharmaceuticals. PMID- 16353090 TI - Investigating the latent polymorphism of maleic acid. AB - The unexpected appearance of a new polymorph of maleic acid is reported and a computational study addresses the predictability of this new polymorph and future potential polymorphism. PMID- 16353091 TI - Non-covalent expansion of an organic bilayer involving exo-cavity interplay of tetraphenylphosphonium with para-sulfonato-calix[4]arene. AB - Reaction of equimolar amounts of sodium para-sulfonato-calix[4]arene, tetraphenylphosphonium chloride and ytterbium chloride in water results in the formation of a mineral clay-like structure, where the hydrophobic tetraphenylphosphonium cations interpose a bilayer arrangement based on a 2D coordination polymer of (calix[4]arene)5-/Na+/Yb3+. PMID- 16353092 TI - Mechanism of alpha-oxoamine synthases: identification of the intermediate Claisen product in the 8-amino-7-oxononanoate synthase reaction. AB - The reactive beta-ketoacid pyridoxal-5'-phosphate aldimine formed in the condensation step of the 8-amino-7-oxononanoate synthase reaction was 'trapped' in the enzyme-bound form by carrying out the reaction with l-alanine methyl ester and pimeloyl-CoA affording the more stable methyl ester of the putative intermediate, the characterisation of which provides the first definitive evidence for a beta-ketoacid intermediate in an alpha-oxamine synthase mechanism. PMID- 16353093 TI - Observation of the formation of supported bilayers by amphiphilic peptidyl-RNA. AB - Amphiphilic peptidyl-RNA conjugates, molecules that mimic natural peptidyl transfer RNA, are capable of self-assembling on glass substrates as vesicles and supported bilayers. PMID- 16353094 TI - Asymmetric organocatalytic conjugate addition of malonates to enones using a proline tetrazole catalyst. AB - 5-Pyrrolidin-2-yltetrazole performs as a useful organocatalyst for the asymmetric addition of malonates to a range of enones, with good to excellent enantioselectivities. PMID- 16353095 TI - Formation of insoluble perylenetetracarboxylic diimide films by electro- or photo crosslinking of pyrrole units. AB - Perylenetetracarboxylic diimide derivatives bearing 2- or 4-peripheral pyrrole pendants could be efficiently crosslinked to form an insoluble film either by electropolymerization or visible light induced oxidative photopolymerization of the pyrrole units. PMID- 16353096 TI - Ionic liquids--media for unique phosphorus chemistry. AB - Ionic liquids have been shown to offer hitherto unseen control as both a storage solvent for PCl3 and POCl3 and reaction media for fluorination and mixed anhydride formation under benign conditions. PMID- 16353097 TI - Fabrication of monodisperse colloidal array with confinement effects. AB - A monodisperse 1D colloidal array is prepared from monomer directly combining precipitation polymerization and confinement effects. PMID- 16353098 TI - Vacuum-ultraviolet ionization spectroscopy of the jet-cooled RNA-base uracil. AB - Adiabatic ionization potential (9.3411 +/- 0.0008 eV) and cationic vibrational structure of the jet-cooled RNA-base uracil are both accurately and precisely determined for the first time using a vacuum-ultraviolet mass-analyzed threshold ionization spectroscopy. PMID- 16353099 TI - From Pd nanoparticles to single crystals: 1,3-butadiene hydrogenation on well defined model catalysts. AB - Although 1,3-butadiene hydrogenation is known to be a structure-sensitive reaction, correlation of the catalytic activity with the exact Pd particle surface structure shows that the reaction is in fact particle size independent. PMID- 16353100 TI - Stabilisation of a paramagnetic BH(4-)-bridged dinickel(II) complex by a macrodinucleating hexaaza-dithiophenolate ligand. AB - The first paramagnetic borohydrido-bridged dinuclear nickel(II) complex, [(L)Ni(II)2(mu(1,3)-BH4)]+, stabilised by a sterically demanding hexaaza dithiophenolate macrocycle, has been obtained by the reaction of [(L)Ni(II)2(micro-ClO4)]+ with N(n)Bu4BH4. PMID- 16353101 TI - Solid-supported chemiluminescence and electrogenerated chemiluminescence based on a tris(2,2'-bipyridyl)ruthenium(II) derivative. AB - We report a simple and efficient technique for the covalent immobilisation of a tris(2,2'-bipyridyl)ruthenium(II) derivative suitable for both chemiluminescence and electrochemiluminescence detection. PMID- 16353102 TI - The first insoluble polymer-bound palladium complexes of 2 pyridyldiphenylphosphine: highly efficient catalysts for the alkoxycarbonylation of terminal alkynes. AB - Palladium complexes of 2-pyridyldiphenylphosphine anchored on polystyrene, polymethylmethacrylate and styrene-methylmethacrylate copolymer form highly active heterogeneous catalysts for the alkoxycarbonylation of terminal alkynes with activities approaching those obtained under homogeneous conditions. PMID- 16353103 TI - Ruthenium biimidazole complexes as anion receptors. AB - The behavior of the compound [RuCl(cym)(H2biim)][BAr'4] (cym = eta6-para isopropylmethylbenzene, Ar' = 3,5-bis(trifluoromethyl)phenyl), synthesized from [[RuCl(cym)]2(mu-Cl)2], H2biim and NaBAr'4, has been studied as a receptor of anions both in solution and in the solid state. PMID- 16353104 TI - From allene to allene: a palladium-catalyzed approach to beta-allenyl butenolides and their application to the synthesis of polysubstituted benzene derivatives. AB - An allene to allene protocol for the synthesis of beta-allenyl butenolides in moderate to high yields from 2,3-allenoic acids and propargylic carbonates catalyzed by Pd(OAc)2-TFP has been developed; the products were applied successfully to the Diels-Alder reaction with electron-deficient alkynes to afford polysubstituted benzene derivatives with an excellent regioselectivity. PMID- 16353105 TI - Methyl transfer reaction from monomethyltin reagent under palladium(0) catalysis: a versatile method for labelling with carbon-11. AB - The 11C-monomethylstannate prepared from [11C]-methyl iodide and Lappert's stannylene, was subject to a palladium-mediated cross-coupling reaction with an aryl halide under ligand-free conditions, to afford easily purified 11C methyl(hetero)arenes in high radiochemical yields. PMID- 16353106 TI - Cofacial porphyrin multilayers via layer-by-layer assembly. AB - This paper reports a new layer-by-layer assembly approach to fabricate multilayers of cofacially aligned porphyrins on solid supports by a selective siloxane formation utilizing tetraphenylporphyrinatosilicon(IV) chloride as the building block. PMID- 16353107 TI - Linkage of N3 dye to N3 dye on nanocrystalline TiO2 through trans-1,2-bis(4 pyridyl)ethylene for enhancement of photocurrent of dye-sensitized solar cells. AB - Linking of N3 dye to another TiO2-attached N3 dye rendered an enhanced short circuit photocurrent and thereby higher efficiency for the dye-sensitized solar cell with the pertinent TiO2 film electrode. PMID- 16353108 TI - A highly selective charge transfer fluoroionophore for Cu2+. AB - A dual fluorescent charge transfer fluoroionophore (1) with its ionophore incorporated in the electron acceptor was developed and was found to show a highly selective fluorescent response to Cu2+ with a dramatic enhancement in its CT emission. PMID- 16353109 TI - Two regioisomeric and exclusively selective Hg(II) sensor molecules composed of a naphthalimide fluorophore and an o-phenylenediamine derived triamide receptor. AB - This communication presents two regioisomeric fluorescent PET Hg(II) sensors RS, which contain a novel o-phenylenediamine based triamide receptor; RS could be used to detect Hg(II) with exclusive selectivity, high fluorescence enhancement, and fast and reversible response in a neutral buffered aqueous solution. PMID- 16353110 TI - Organ donation and transplantation: a lesson from Mr. Green. PMID- 16353111 TI - Why focus on implant infections? PMID- 16353112 TI - Biofilm in implant infections: its production and regulation. AB - A significant proportion of medical implants become the focus of a device-related infection, difficult to eradicate because bacteria that cause these infections live in well-developed biofilms. Biofilm is a microbial derived sessile community characterized by cells that are irreversibly attached to a substratum or interface to each other, embedded in a matrix of extracellular polymeric substances that they have produced. Bacterial adherence and biofilm production proceed in two steps: first, an attachment to a surface and, second, a cell-to cell adhesion, with pluristratification of bacteria onto the artificial surface. The first step requires the mediation of bacterial surface proteins, the cardinal of which is similar to S. aureus autolysin and is denominated AtlE. In staphylococci the matrix of extracellular polymeric substances of biofilm is a polymer of beta-1,6-linked N-acetylglucosamine (PIA), whose synthesis is mediated by the ica operon. Biofilm formation is partially controlled by quorum sensing, an interbacterial communication mechanism dependent on population density. The principal implants that can be compromised by biofilm associated infections are: central venous catheters, heart valves, ventricular assist devices, coronary stents, neurosurgical ventricular shunts, implantable neurological stimulators, arthro-prostheses, fracture-fixation devices, inflatable penile implants, breast implants, cochlear implants, intraocular lenses, dental implants. Biofilms play an important role in the spread of antibiotic resistance. Within the high dense bacterial population, efficient horizontal transfer of resistance and virulence genes takes place. In the future, treatments that inhibit the transcription of biofilm controlling genes might be a successful strategy in inhibiting these infections.A significant proportion of medical implants become the focus of a device-related infection, difficult to eradicate because bacteria that cause these infections live in well-developed biofilms. Biofilm is a microbial derived sessile community characterized by cells that are irreversibly attached to a substratum or interface to each other, embedded in a matrix of extracellular polymeric substances that they have produced. Bacterial adherence and biofilm production proceed in two steps: first, an attachment to a surface and, second, a cell-to-cell adhesion, with pluristratification of bacteria onto the artificial surface. The first step requires the mediation of bacterial surface proteins, the cardinal of which is similar to S. aureus autolysin and is denominated AtlE. In staphylococci the matrix of extracellular polymeric substances of biofilm is a polymer of beta-1,6-linked N-acetylglucosamine (PIA), whose synthesis is mediated by the ica operon. Biofilm formation is partially controlled by quorum sensing, an interbacterial communication mechanism dependent on population density. The principal implants that can be compromised by biofilm associated infections are: central venous catheters, heart valves, ventricular assist devices, coronary stents, neurosurgical ventricular shunts, implantable neurological stimulators, arthro-prostheses, fracture-fixation devices, inflatable penile implants, breast implants, cochlear implants, intra-ocular lenses, dental implants. Biofilms play an important role in the spread of antibiotic resistance. Within the high dense bacterial population, efficient horizontal transfer of resistance and virulence genes takes place. In the future, treatments that inhibit the transcription of biofilm controlling genes might be a successful strategy in inhibiting these infections. PMID- 16353113 TI - Molecular genetics of Staphylococcus epidermidis biofilms on indwelling medical devices. AB - Staphylococcus epidermidis is an opportunistic pathogen associated with foreign body infections and nosocomial sepsis. The pathogenicity of S. epidermidis is mostly due to its ability to colonize indwelling polymeric devices and form a thick, multilayered biofilm. Biofilm formation is a major problem in treating S. epidermidis infection as biofilms provide significant resistance to antibiotics and to components of the innate host defenses. Various cell surface associated bacterial factors play a role in adherence and accumulation of the biofilm such as the polysaccharide intercellular adhesin and the autolysin AtlE. Furthermore, recent studies have shown that global regulators such as the agr quorum sensing system, the transcriptional regulator sarA and the alternative sigma factor sigB have an important function in the regulation of biofilm formation. Understanding the many complex mechanisms involved in biofilm formation is a key factor in the search for new anti-staphylococcal therapeutics. PMID- 16353114 TI - Implant infections due to enterococci: role of capsular polysaccharides and biofilm. AB - Enterococci are natural inhabitants of the gastrointestinal tract and of the female genital tract of humans and many animals. In recent years, enterococci have been increasingly recognized as important human pathogens causing infections associated with medical devices. Their resistance to most antimicrobial agents and their ability to form biofilm has contributed to the increasing incidence of nosocomial enterococcal infections. Enterococci possess a capsular polysaccharide composed of a glycerol-teichoic acid-like molecule consisting of repeating units of 6-alfa-D-glucose-1-2-glycerol-3-PO4 , substituted on carbon 2 with a alfa-2,1 linked molecule of glucose. Using both immunologic and genetic data E. faecalis can be assigned to specific serotypes based on capsular polysaccharides. Clinical examples of foreign-body infections due to enterococci are described, comprising infections of artificial joints, implanted intravascular catheters, artificial hearts and artificial valves, stents, liquor shunt devices, and intraocular infections. Methods to prevent and/or treat enterococcal infections are presented. PMID- 16353115 TI - Etiology of implant orthopedic infections: a survey on 1027 clinical isolates. AB - In spite of the recent achievements derived from modern protocols of prophylaxis, orthopedic surgical infections still remain unacceptably frequent, especially in light of the often devastating outcomes of septic complications. The spectrum and the prevalence of the bacteria most frequently involved in orthopedic infections are here explored, with particular reference to those infections associated to implant biomaterials, which were grouped based on device typology. During a 30 months period (from September 2000 to April 2003), 1027 microbial strains were consecutively isolated from 699 patients undergoing revision surgery at the Rizzoli Orthopedic Institute. 775 (75.5%) of all these microorganisms were identified as belonging to the Staphylococcus genus, 82 (8%) to the Enterobacteriaceae family, 75 (7.3%) to the Pseudomonas genus, 54 (5.3%) to the Enterococcus genus and 20 (1.9%) to the Streptococcus genus. While confirming the importance of staphylococci as the most diffuse cause of infection, our data indicate an unexpectedly high prevalence of S. epidermidis on infected hip and knee arthroprostheses, respectively of 42% and 44%. The spectrum of bacteria infecting either internal or external fracture fixation devices appears to differ from that of hip and knee arthroprostheses and more closely resembles that of infections non-associated to medical devices, being characterized by a relatively higher prevalence of Staphylococcus aureus (over 40%) and Pseudomonas aeruginosa. Enterobacteriaceae and members of the Streptococcus and Corynebacterium genera are frequently associated with implants in which surgical incisions were made near the perineum, determining a completely altered spectrum. PMID- 16353116 TI - Pathogenesis of implant infections by enterococci. AB - Enterococci are commensals of human and animal intestinal tract that have emerged in the last decades as a major cause of nosocomial infections of bloodstream, urinary tract and in infected surgical sites. Enterococcus faecalis is responsible for ca. 80% of all enterococcal infections while Enterococcus faecium accounts for most of the others; among the most relevant risk factors for development of enterococcal infections is the presence of implanted devices. The pathogenesis of such infections is poorly understood, but several virulence factors have been proposed. Among them, the ability to form biofilm has recently been shown to be one of the most prominent features of this microorganism, allowing colonization of inert and biological surfaces, while protecting against antimicrobial substances, and mediating adhesion and invasion of host cells and survival within professional phagocytes. Biofilm formation has been shown to be particularly important in the development of prosthetic valve enterococcal endocarditis and stent occlusion. Enterococci are also able to express other surface factors that may support colonization of both inert and biological surfaces, and that may be involved in the invasion of, and survival within, the host cell. PMID- 16353117 TI - Clinical behavior of implant infections due to Staphylococcus epidermidis. AB - Surgical implants and other foreign material are increasingly used in modern medicine to restore or to improve the function of the human body. Infection of an implant is associated with considerable morbidity due to frequent hospitalizations, surgery and antimicrobial treatment. The underlying mechanism is the formation of a bacterial biofilm on the surface of the implanted body. The recognition and diagnosis of implant infections is essential for further therapy and, above all, the decision to remove and exchange the implant. METHODS: We compared the data of 60 patients with implant infections with those of 60 patients with transient bacteremia caused by Staphylococcus epidermidis. The pathogens isolated from blood were characterized with regard to antimicrobial susceptibility and formation of biofilms using a static microtiter plate model. Wild type skin isolates from non-hospitalized healthy volunteers served as control with regard to antimicrobial susceptibility and biofilm formation. RESULTS: Clinical signs and symptoms, underlying diseases and outcome were not different in either group. However, patients with implant infection had fever over a longer time (mean 12 days versus 3 days, respectively, p < 0.05) and more often positive blood cultures than patients with transient bacteremia (3.1 versus 1.2, p < 0.05). Thrombocytopenia was observed in patients with implant infections but not in patients with transient bacteremia (p < 0.05). Biofilms were formed in 86.4 % of the isolates in implant infection, in 88.8 % in transient bacteremia and in 76.9 % of the isolates from healthy volunteers (not significant). Multi resistance to penicillin, oxacillin, erythromycin, clindamycin, ciprofloxacin and trimethoprim was more common in the hospital strains than in the wild type strains (75.6 % versus 48.7 %, p < 0.05). CONCLUSIONS: The clinical features of implant infections are indistinguishable from those of transient bacteremia. Persisting fever and multiple blood culture yielding the growth of skin flora bacteria are strong indicators for infection of implanted material. Biofilm formation and antimicrobial multi-resistance, as common in implant infection as in transient bacteremia, seem to be accessory factors in infections due to Staphylococcus epidermidis. PMID- 16353118 TI - Implant infection and infection resistant materials: a mini review. AB - Implant infection is an aggressive, often irreducible post-surgical infection. It remains the primary cause of implant failure. Bacterial contamination during surgery and subsequent adhesion onto biomaterial surface of opportunistic microorganisms, such as staphylococcal species, exopolysaccharidic slimes or specific adhesins, initiates the implant infection. Pathogenesis of periprosthestic infection is the focus of studies aimed at developing infection resistant materials. PMID- 16353119 TI - Effects of sterilization on implant mechanical property and biocompatibility. AB - This article concisely reviews the effects of sterilization on the mechanical properties and surface chemistries of implantable biomaterials. This article also summarizes the biological effects of the sterilization-related changes in the implant. Because there are so many different types of implant materials currently in use (including metals, polymers, and diverse biological materials), the response of tissue to these different materials varies dramatically. This review further discusses the effects of sterilization on in vivo and in vitro tissue response specifically to implantable metals and polyethylene, with the possibility of future biocompatibility testing of the implants sterilized with supercritical phase carbon dioxide sterilization. PMID- 16353120 TI - Factors affecting microbial adhesion to stainless steel and other materials used in medical devices. AB - The role of biofilm in medical device associated infections is well documented. Biofilms are more resistant to antibiotics than planktonic cells, these are extremely difficult to treat. Prevention strategies include efforts to insert implants under stringent aseptic conditions, and also encompass the development of novel materials which interfere with the initial attachment of microorganisms to the surface of the device. Microbial cells also attach onto hygienic surfaces in the hospital setting, and thereby pose a cross-infection problem. In this case, vigorous cleaning and sanitizing regimes may be employed in addition to any surface modifications. Many factors affect the initial attachment of organisms to inert substrata, and their subsequent retention or removal/detachment, including the physical and chemical nature and location of the substratum, the type of organic material and microorganisms potentially fouling the surface, and the nature of the interface (solid-liquid in the body; solid-air on environmental surfaces). Focusing on one factor, surface topography, it is apparent that many further variables need to be defined in order to fully understand the interactions occurring between the cell and surface. It is therefore important when modifying one substratum surface property in order to reduce adhesion, to also consider other potentially confounding factors. PMID- 16353121 TI - Modern strategies in the prevention of implant-associated infections. AB - The application of medical devices either for temporary or permanent use has become an indispensible part of almost all fields of medicine. However, foreign bodies are associated with a substantial risk of bacterial and fungal infections. Implant-associated infections significantly contribute to the still increasing problem of nosocomial infections. To reduce the incidence of such infections, specific guidelines providing evidence-based recommendations and comprising both technological and nontechnological strategies for prevention have been established. Strict adherence to hygienic rules during insertion or implantation of the device are aspects of particular importance. Besides such basic and indispensable aspects, the development of new materials which could withstand microbial adherence and colonization has become a major topic in recent years. Modification of surface by primarily physico-chemical methods may lead to a change in specific and unspecific interactions with microorganisms and, thus, to a reduction in microbial adherence. Medical devices made out of a material that would be ideally antiadhesive or at least colonization-resistant would be the most suitable candidates to avoid colonization and subsequent infection. However, it appears impossible to create a surface with an absolute 'zero'-adherence due to thermodynamical reasons and due to the fact that a modified material surface is in vivo rapidly covered by plasma and connective tissue proteins. Therefore, another concept for the prevention of implant-associated infections involves the impregnation of devices with various antimicrobial substances such as antibiotics, antiseptics, and/or metals. In fact, already commercially available materials for clinical use such as antimicrobial catheters have been introduced, in part with considerable impact on subsequent infections. However, future studies are warranted to translate the knowledge on the pathogenesis of device associated infections into applicable prevention strategies. PMID- 16353122 TI - Vaccines and immunotherapy for staphylococcal infections. AB - Nosocomial or hospital-acquired infections are associated with prolonged hospitalizations and increased healthcare costs. Infections associated with surgical implants are becoming more difficult and more costly to manage, as they require repeated surgical procedures and a longer period of time to treat patients. Continued advances in the use of medical devices, an increase in the number of immunocompromised patients, and a steady rise in the prevalence of antibiotic-resistant organisms has renewed interest in the development of novel therapies that can be used to prevent and treat nosocomial infections. This review provides an overview of bacterial adhesins and focuses on novel immunological therapies developed to treat staphylococcal infections. PMID- 16353123 TI - Nanocapsules as drug delivery systems. AB - Dispersed polymer nanocapsules can serve as nano-sized drug carriers to achieve controlled release as well as efficient drug targeting. The dispersion stability and the primary physiological response are mainly determined by the type of the surfactant and the nature of the outer coating. Their release and degradation properties largely depend on the composition and the structure of the capsule walls. Another important criterion is the capsule size, where an optimum is generally seen for radii ranging between 100 and 500 nm. Nanocapsules can be prepared by four principally different approaches: interfacial polymerization, interfacial precipitation, interfacial deposition, and self assembly procedures. All these procedures offer their individual advantages and disadvantages when it comes to the design of optimized drug carrier systems. The most important capsule parameters such as capsule radius distribution, the capsule surface, the thickness and the permeability of the capsule membrane and its thermal or chemical decomposition, are discussed and examples are shown. In combination with efficient preparation procedures, nanocapsule dispersions allow for new and promising approaches in many kinds of pharmaceutical therapies. PMID- 16353124 TI - Implant-associated posttraumatic osteomyelitis: collateral damage by local host defense? AB - Infections following osteosynthesis or total joint replacement, also known as ''implant-associated posttraumatic osteomyelitis'', represent a major complication in orthopedic and trauma surgery. While the formation of bacterial biofilms on the implanted osteosynthesis materials is generally accepted as cause of the persistent infection, the molecular mechanisms leading to the progressive and destructive local inflammatory process and eventually to bone degradation, the osteolysis, have not been delineated. Here we provide evidence supporting the hypothesis that it is not the infection per se that causes tissue degradation and osteolysis, but rather the cytotoxic, proteolytic, and proinflammatory effector functions of cells of the host defense, particularly of the infiltrating polymorphonuclear neutrophils. PMID- 16353125 TI - Influence of sub-inhibitory concentrations of antimicrobial agents on biofilm formation in indwelling medical devices. AB - Biofilms of Staphylococcus epidermidis and Candida spp. are two of the most frequent factors of infections associated with the use of indwelling medical devices. Several strategies have been proposed and/or developed to prevent infection. The aim of this study was to compare the effect of sub-inhibitory concentrations of anti-microbial agents on biofilm formation. Biofilms of three strains of S. epidermidis and two of both Candida albicans and Candida dubliniensis were formed in the presence of three antibiotics and two antifungal agents respectively. Based on the control samples, the percentage of biofilm formation inhibition by the different agents was determined and compared. The results showed that the influence of the antibacterial and antifungal agents tested is strain dependent, with the effect of the different agents also varying among strains, even though they have the same mechanism of action. PMID- 16353126 TI - Antibiotic resistance in Staphylococcus aureus and Staphylococcus epidermidis clinical isolates from implant orthopedic infections. AB - In the last decade the rising phenomenon of resistance to most common antibiotic drugs among staphylococcal clinical isolates has been a reason for serious concern and alarm. The present study investigated the prevalence of antibiotic resistance within a large microbial collection including 530 clinical strains of S. aureus and 408 strains of S. epidermidis to a panel of 16 different drugs. All strains were isolated from orthopedic infections, either associated or non associated with implant materials. Interestingly, our data show that the profile of the prevalence of antibiotic resistance within the two species of pathogens is extremely similar for the vast majority of the drugs screened. The only statistically significant variations in prevalence concerned, in order of relevance, the following 5 out of 16 antibiotics: sulfamethoxazole (in combination with trimethoprim), erythromycin, and, to a lesser extent, oxacillin, imipenem, and clindamycin. In the case of Staphylococcus aureus, the isolates associated to implant materials were found more frequently resistant to all 4 aminoglycosides screened as well as to ciprofloxacin. PMID- 16353127 TI - Electrospray ionization quadrupole time-of-flight tandem mass spectrometric analysis of hexamethylenediamine-modified maltodextrin and dextran. AB - A combined methodology for obtaining at the preparative scale and characterization by nanoelectrospray ionization (nanoESI) quadrupole time-of flight (QTOF) mass spectrometry (MS) and tandem MS (MS/MS) of linear polysaccharides modified at the reducing end is presented. Two polydisperse maltodextrins (1000 and 3000 Da) and a high molecular weight polydisperse dextran (6000 Da) were coupled with hexamethylenediamine (HMD). The coupling products were analyzed by nanoESI-QTOF-MS in the positive ion mode and MS/MS using collision-induced dissociation (CID) at low energies. In the HMD-M1000 mixture, the polysaccharide chains containing from 2 to 8 Glc residues were detected, while in HMD-M3000 we identified a complete series of chains containing from 8 to 21 Glc moieties. The employed ESI conditions enhanced the detection of chains with up to 46 Glc residues in the HMD-D6000 sample. By optimized MS/MS, HMD modified polysaccharides of 3, 4, 5, 12 and 46 degrees of polymerization yielded product ion spectra exhibiting the whole set of Y- and B-fragment ions. The MS structural data were obtained within a few minutes of signal acquisition, with a sample consumption situating the analysis sensitivity in the picomolar range. PMID- 16353128 TI - Differentiation of diiodothyronines using electrospray ionization tandem mass spectrometry. AB - Diiodothyronines 3,5-diiodothyronine (3,5-T2), 3',5'-diiodothyronine (3',5'-T2), and 3,3'-diiodothyronine (3,3'-T2) are important metabolites of 3,5,3' triiodothyronine (T3) and 3,3',5'-triiodothyronine (rT3; reverse T3). In this paper, a novel and rapid method for identifying and quantifying 3,5-T2, 3',5'-T2 and 3,3'-T2 has been introduced using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Fragmentation patterns were proposed on the basis of our data obtained by ESI-MS/MS. MS2 spectra in either negative ionization mode or positive ionization mode can be used to differentiate 3,5-T2, 3',5'-T2 and 3,3' T2. On the basis of the relative abundance of fragment ions in MS2 spectra under the positive ionization mode, quantification of the 3,5-T2, 3',5'-T2 and 3,3'-T2 isomers in mixtures is also achieved without prior separation. PMID- 16353129 TI - Investigation of intramolecular proton migration in a series of model, metal cationized tripeptides using in situ generation of an isotope label. AB - In this study we used an isotope label, generated in situ, to investigate intramolecular proton migration or scrambling during formation of [b(2)+17+Li](+) products by collision-induced dissociation (CID) of Li(+)-cationized tripeptides. To generate the isotope label, we used a McLafferty-type rearrangement of N terminally acetylated, C-terminal peptide tert-butyl esters in which all amide positions were exchanged with deuterium. Using a set of small, model peptides, we show that intramolecular proton scrambling occurs during CID, particularly amongst adjacent sites along a peptide backbone, on the time scales employed for low-energy collisional activation in an ion-trap mass spectrometer. PMID- 16353130 TI - A new hybrid electrospray Fourier transform mass spectrometer: design and performance characteristics. AB - A new hybrid electrospray quadrupole Fourier transform mass spectrometry (FTMS) instrument design is shown and characterized. This instrument involves coupling an electrospray source and mass-resolving quadrupole, ion accumulation, and collision cell linear ion trap system developed by MDS Sciex with a home-built ion guide and ion cyclotron resonance (ICR) cell. The iterative progression of this design is shown. The final design involves a set of hexapole ion guides to transfer the ions from the accumulation/collision trap through the magnetic field gradient and into the cell. These hexapole ion guides are separated by a thin gate valve and two conduction limits to maintain the required <10(-9) mbar vacuum for FTICR. Low-attomole detection limits for a pure peptide are shown, 220 000 resolving power in broadband mode and 820 000 resolving power in narrow-band mode are demonstrated, and mass accuracy in the <2 ppm range is routinely available provided the signal is abundant, cleanly resolved, and internally calibrated. This instrument design provides high experimental flexibility, allowing Q2 CAD, SORI-CAD, IRMPD, and ECD experiments with selected ion accumulation as well as experiments such as nozzle skimmer dissociation. Initial top-down mass spectrometry experiments on a protein is shown using ECD. PMID- 16353131 TI - Expression and functional analysis of Bax inhibitor-1 in human breast cancer cells. AB - Recently, deregulated expression of the anti-apoptotic protein Bax inhibitor-1 (BI-1) has been shown in several human cancers. In this report, we show that BI-1 is expressed at various levels in six different human breast cancer cell lines. In order to investigate the function of BI-1 in oestrogen-dependent MCF-7, T-47D and oestrogen-independent MDA-MB-231 breast cancer cells, the RNA interference technique was used to knock down BI-1 expression specifically. Suppression of BI 1 expression caused a significant increase in spontaneous apoptosis in MDA-MB-231 cells, whereas MCF-7 and T-47D cells remained almost unaffected. Furthermore, BI 1 expression analysis using a cancer profiling array showed up-regulation of BI-1 expression in cancer samples of breast, uterus and ovary, whereas down-regulated BI-1 expression was identified in stomach, colon, kidney, lung and rectal cancer. In addition, immunohistochemical studies using a BI-1-specific antibody on human breast cancer specimens also revealed that BI-1 is expressed in the majority of cases. Moreover, to analyse whether BI-1 expression is oestrogen receptor dependent, tumour cells were treated with oestradiol, ICI and tamoxifen: this showed no significant changes in BI-1 expression. Taken together, our results demonstrate that BI-1 expression is differentially deregulated in different cancers and that BI-1 plays an important role in preventing certain breast cancer cells from undergoing apoptosis. Thus, the development of novel therapeutic strategies based on targeting BI-1 gene expression in breast cancer could be restricted to selected individual cancer types. PMID- 16353132 TI - Immunoglobulin VH genes in thymic MALT lymphoma are biased toward a restricted repertoire and are frequently unmutated. AB - Thymic MALT lymphoma shows certain distinctive features among MALT lymphomas, such as expression of IgA isotype, consistent lack of API2-MALT1 gene fusion, and very strong association with autoimmune disease, especially Sjogren's syndrome. To help clarify the nature of the clonal lymphoid infiltrates, we analysed the usage and somatic hypermutation of the Ig heavy chain variable region (V(H)) genes in nine different cases. The V(H) rearrangement was potentially functional in all cases and was restricted to the V(H)3 family. V(H) usage was biased toward V(H)3-30 (five cases) and V(H)3-23 (three cases) segments, which have both been frequently expressed by autoimmune B cells. Somatic hypermutation was absent in five cases. Fewer than the expected replacement mutations were found in the framework regions in two cases, indicating a negative antigen selection pressure. Ongoing mutation was absent in all cases. D segment usage was varied, whereas J(H) segment usage was restricted to J(H)4. The observed patterns of V(H) usage and mutations suggested that specific antigens may play a pathologically relevant role in the genesis or progression of thymic MALT lymphoma. PMID- 16353133 TI - Enumeration of peripheral lymphocyte subsets using 6 vs. 4 color staining: a clinical evaluation of a new flowcytometer. AB - Technological advances in instruments allow the evaluation of many lymphocyte subsets in one step. The aim of this study was to evaluate the new FACSCanto flowcytometer in routine conditions, using a 6 color combination, single platform, whole blood, lysis, no wash protocol. METHODS: Two systems were simultaneously compared on 67 blood samples and external quality controls, using CD3,CD4, CD8, CD19, CD16/56, and CD45 in one tube TRUCOUNT beads (BD Biosciences) or two tubes (TetraChrome and Flowcount, Beckman-Coulter and DakoCytomation). RESULTS: The day-to-day instrument detection but automatic compensations were stable. Manual compensation settings were satisfactory using available facilities. Commercial and UK NEQAS quality control results were acceptable. The intra-experiment reproducibility was good (coefficient of variation (CV)<3%) but highly operator-dependent (CD4+ T cell count CVs from 1.2 to 9.7, six operators). Storage of samples was acceptable, but storage of stained samples altered absolute count reliability. Serial dilutions show a good count accuracy. The FACScanto T subsets and B cell data were highly correlated with our reference values (r2>0.87) and absolutes values were very close (slopes>0.89). The gating strategy, fluorochrome choice, and compensation setting are discussed. A few improvements are expected (sample loader, data management, auto-gating, acquisition parameters, sample mixing, absolute values calculation, etc). In conclusions, despite its complexity, 6 color staining is a reliable, stable, and highly informative technique for lymphocyte subset monitoring but remains to be optimized. PMID- 16353134 TI - Age of diagnosis of colorectal cancer in HNPCC patients is more complex than that predicted by R72P polymorphism in TP53. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominantly inherited cancer syndrome associated with germline mutations in DNA mismatch repair (MMR) genes. Recently a polymorphism at codon 72 (R72P) in the tumour suppressor gene TP53 has been implicated in the age of disease onset in HNPCC. In this report we have studied a large cohort of HNPCC patients to assess the impact of this polymorphism on disease expression and age of diagnosis of colorectal cancer (CRC). DNA samples from 218 HNPCC mutation positive patients from Australia and Poland were genotyped for the arginine to proline change at codon 72 in the TP53 gene. The association between the polymorphism and disease characteristics (mutation status, disease expression and age of diagnosis of CRC) was tested using Pearson's Chi-square and Kaplan-Meier survival analysis. Our study of Australian and Polish HNPCC patients does not provide evidence for an association between the Arg/Pro (GC) genotype of the R72P polymorphism and age of diagnosis of CRC. The R72P polymorphism was examined in HNPCC patients and found to be not associated with disease development in either the Australian or Polish populations. When gene mutation status (hMLH1 or hMSH2) was included in the analysis some evidence of an affect was observed. The genotyping revealed in the Australian population that the R72P polymorphism was under-represented in the hMSH2 group whereas it was over-represented in the Polish hMSH2 group. A similar trend was observed for hMLH1 in both groups but was not significant. Age of diagnosis of CRC in HNPCC patients is therefore more complex than that predicted by the R72P TP53 polymorphism alone, suggesting an inter-relationship with other genetic and/or environmental factors. PMID- 16353135 TI - Neuroblastoma cell death in response to docosahexaenoic acid: sensitization to chemotherapy and arsenic-induced oxidative stress. AB - Docosahexaenoic acid (DHA) is an omega-3 polyunsaturated fatty acid vital for the developing nervous system and significantly decreased in neuroblastoma cells compared to nontransformed nervous tissue. We investigated whether supplementation of DHA affects the susceptibility of neuroblastoma cells to oxidative stress generated endogenously and in response to cytotoxic therapy. DHA, but not the monounsaturated oleic acid (OA), induced dose- and time dependent neuroblastoma cell death. DHA supplementation was associated with depolarization of the mitochondrial membrane potential, production of reactive oxygen species (ROS) and accumulation of DNA in sub-G1 phase of the cell cycle. The antioxidant, vitamin E, inhibited mitochondrial depolarization and subsequent cell death induced by DHA, whereas, the mitochondrial pore inhibitor, cyclosporin A, partly inhibited DHA-induced neuroblastoma cell death. Depletion of glutathione by L-buthionine-sulfoximine significantly enhanced the cytotoxic effects of DHA. Nontransformed fibroblasts were not substantially affected by DHA. DHA, but not OA, significantly enhanced the cytotoxicity of cisplatin, doxorubicin and irinotecan both in chemosensitive and in multidrug-resistant neuroblastoma cells. DHA potently sensitized neuroblastoma cells to a clinically relevant concentration (1 microM) of arsenic trioxide (As2O3) and enhanced the effect of the nonsteroidal antiinflammatory drug (NSAID), diclofenac. These findings provide experimental evidence that the omega-3 fatty acid, DHA, is cytotoxic to drug-resistant neuroblastoma. The potent action of DHA with arsenic trioxide, NSAID and chemotherapeutic agents suggests clinical testing of this therapeutic concept in children with neuroblastoma. PMID- 16353136 TI - Genome-wide gene expression profiling of cervical cancer in Hong Kong women by oligonucleotide microarray. AB - An analysis of gene expression profiles obtained from cervical cancers was performed to find those genes most aberrantly expressed. Total RNA was prepared from 29 samples of cervical squamous cell carcinoma and 18 control samples, and hybridized to Affymetrix oligonucleotide microarrays with probe sets complementary to over 20,000 transcripts. Unsupervised hierarchical clustering of the expression data readily distinguished normal cervix from cancer. Supervised analysis of gene expression data identified 98 and 139 genes that exhibited >2 fold upregulation and >2-fold downregulation, respectively, in cervical cancer compared to normal cervix. Several of the genes that were differentially regulated included SPP1 (Osteopontin), CDKN2A (p16), RPL39L, Clorf1, MAL, p11, ARS and NICE-1. These were validated by quantitative RT-PCR on an independent set of cancer and control specimens. Gene Ontology analysis showed that the list of differentially expressed genes included ones that were involved in multiple biological processes, including cell proliferation, cell cycle and protein catabolism. Immunohistochemical staining of cancer specimens further confirmed differential expression of SPP1 in cervical cancer cells vs. nontumor cells. In addition, 2 genes, CTGF and RGS1 were found to be upregulated in late stage cancer compared to early stage cancer, suggesting that they might be involved in cancer progression. The pathway analysis of expression data showed that the SPP1, VEGF, CDC2 and CKS2 genes were coordinately differentially regulated between cancer and normal. The present study is promising and provides potential new insights into the extent of expression differences underlying the development and progression of cervical squamous cell cancer. This study has also revealed several genes that may be highly attractive candidate molecular markers/targets for cervical cancer diagnosis, prognosis and therapy. PMID- 16353137 TI - Switch to full-length of XAF1 mRNA expression in prostate cancer cells by the DNA methylation inhibitor. AB - X-linked inhibitor of apoptosis protein (XIAP) suppresses apoptotic cell death by binding to caspases and inhibiting their functions, while the XIAP-associated factor1 (XAF1), a zinc finger protein, antagonizes XIAP activities, thereby promoting apoptosis. The aberrant silence of the XAF1 gene has recently been found in various types of cancer cells, which is suggested to be one of the potential mechanisms underlying survival advantages of malignant cells. In the present study, we investigated the XAF1 expression in prostate cancer cells. Compared with normal tissues where a full-length of XAF1 mRNA is predominant, LNCaP and DU145 prostate cancer cell lines only expressed a short form of XAF1 transcripts, whereas PC3 cells exhibited a complete silence of the XAF1 gene. Inhibition of DNA methylation led to a switch to the full length of XAF1 mRNA expression in LNCaP and DU145 cells. The down-regulation of XAF1 expression was also observed in 6/8 tumor samples derived from patients with prostate cancer. Our findings suggest that splicing alterations or downregulation of the XAF1 transcript may occur during the development of prostate cancers due to the aberrant DNA methylation. The alternative splicing of XAF1 mRNA leads to formation of a truncated XAF1 protein with 19 amino acid deletion in its zinc finger domain, which likely affects its functional interaction with XIAP, and consequently, contributes to the pathogenesis of prostate cancers by disrupting balance of the apoptosis machinery. PMID- 16353138 TI - Induction of strong and persistent MelanA/MART-1-specific immune responses by adjuvant dendritic cell-based vaccination of stage II melanoma patients. AB - A significant percentage of stage II melanoma patients (tumor thickness>1 mm) remain at risk of tumor recurrence after primary tumor excision. In this study, we used tumor antigen-pulsed dendritic cells as an adjuvant for immunization of these "high-risk" melanoma patients after resection of the primary tumor. A total of 13 patients were included and vaccinated 6 times every 14 days with autologous dendritic cells pulsed with a MelanA/MART-1 peptide in combination with a recall antigen. Antigen-specific immune responses were monitored before, during and up to 1 year after the last vaccination. The majority of patients exhibited increased recall antigen-specific CD4+ T cell responses upon vaccination. MelanA/MART-1-specific CD8+ T cells were expanded in 9/13 patients resulting in increased frequencies of memory cells in these patients. CD8+ T cells acquired the capacity to secrete IFN-gamma, to proliferate in culture in response to the tumor antigen used for vaccination and postvaccine samples contained MelanA/MART 1-specific T cells that recognized also the natural MelanA/MART-1-antigen expressed by tumor cells. Moreover, vaccination induced a long-lived tumor antigen-specific DTH-reactivity in the majority of the patients, detectable even 12 months after the last immunization. These data demonstrate for the first time that vaccination with tumor antigen-pulsed dendritic cells in a clinically adjuvant setting induces strong and persistent antigen-specific T-cell responses in tumor-free stage II melanoma patients, suggesting that tumor protective T cell immunity can be achieved. PMID- 16353139 TI - GADD45alpha is highly expressed in pancreatic ductal adenocarcinoma cells and required for tumor cell viability. AB - Pancreatic ductal adenocarcinoma is one of the most common causes of cancer death in the western civilization. Recently, NF-kappaB has been shown to be activated in pancreatic ductal adenocarcinoma through constitutive activation of IkappaB kinase (IKK). Inhibition of NF-kappaB by a super-inhibitor of NF-kappaB--delta-N IkappaBalpha--resulted in impaired proliferation and induction of apoptosis, suggesting an important role of NF-kappaB in pancreatic tumorigenesis. Downstream target genes of IkappaBalpha have not been elucidated in pancreatic ductal adenocarcinoma in detail. Using expression profiling by cDNA array analysis of pancreatic ductal adenocarcinoma cell lines stably transfected with super IkappaBalpha, we identified GADD45alpha as a significant regulated gene. GADD45alpha is overexpressed in pancreatic ductal adenocarcinoma at the mRNA and protein level. Using RNAi we show that downregulation of GADD45alpha reduces proliferation and induces apoptosis in pancreatic cancer cells. These findings provide evidence that GADD45alpha contributes to pancreatic cancer cell proliferation and viability. PMID- 16353140 TI - Prophylactic effect of pemirolast, an antiallergic agent, against hypersensitivity reactions to paclitaxel in patients with ovarian cancer. AB - We have previously shown that sensory nerve peptides contribute to the pathogenesis of pulmonary hypersensitivity reactions (HSRs) to paclitaxel in rats. Moreover, pemirolast, an antiallergic agent, reverses the HSRs to paclitaxel, although the mechanism is considered to result from the blockade of paclitaxel-induced release of sensory peptides, rather than the inhibition of histamine release. In the present study, we investigated the preventive effect of pemirolast against acute HSRs in a total of 84 patients who undertook postoperative paclitaxel plus carboplatin chemotherapy every 4 weeks for ovarian cancer. Patients were assigned to receive oral lactose (placebo) or pemirolast (10 mg), 2 hr before paclitaxel infusion. All patients received conventional premedication, including oral diphenhydramine, intravenous ranitidine and intravenous dexamethasone, 30 min before paclitaxel infusion. The HSRs that led to the discontinuance of paclitaxel infusion (grade>or=2) occurred in 5 of 42 patients in placebo group, whereas none of pemirolast-treated 42 patients showed any signs of HSRs. Plasma histamine concentrations were not changed after paclitaxel infusion in either group. Our present findings suggest that pemirolast is potentially useful for prophylaxis of paclitaxel-induced HSRs. In this respect, the use of pemirolast as premedication is expected to be beneficial to the safety management in patients who undergo chemotherapy containing paclitaxel. PMID- 16353141 TI - Detection rates of high-grade prostate cancer during subsequent screening visits. Results of the European Randomized Screening Study for Prostate Cancer. AB - Screening for prostate cancer using prostate-specific antigen (PSA) tests has led to a stage and grade shift as compared to the pre-PSA era. Effectiveness of screening for prostate cancer should be manifested by a reduction in detection rate of aggressive cancers during subsequent screening. In 6 centers of the European Randomized Screening study for Prostate Cancer, a total of 58,710 men were tested for prostate cancer. Screening centers differed with regard to age range, screening interval and biopsy indications. During the 2nd visit, the proportion of Gleason score 6 cancers increased from 62.5 to 75%, mainly at the expense of Gleason score 7 cancers. High-grade (Gleason score 8-10) cancer detection rates varied per screening center during the 1st visit from 5.1 to 41.1, and during the 2nd visit from 6.4 to 29.3/10,000 men. The overall detection rate of high-grade cancers showed a reduction during the 2nd visit from 26 to 12/10,000 men, an effect mainly attributable to the screening center with the highest cancer detection rate (i.e. 507/10,000 men). Variations in detection rates among screening centers related among others to biopsy compliance and age range. PMID- 16353142 TI - Combination of a monoclonal anti-phosphatidylserine antibody with gemcitabine strongly inhibits the growth and metastasis of orthotopic pancreatic tumors in mice. AB - Pancreatic cancer continues to have a dismal prognosis and novel therapy is needed. In this study, we evaluate a promising new target for therapy, phosphatidylserine (PS). PS is an anionic phospholipid located normally on the inner leaflet of the plasma membrane in mammalian cells. In the tumor microenvironment, PS becomes externalized on vascular endothelium. The monoclonal antibody 3G4 binds PS and promotes an inflammatory response against tumor blood vessels, resulting in reduction of tumor growth. Mice with orthotopic pancreatic tumors were treated with 3G4, gemcitabine or a combination of both drugs. Tumor burden including pancreas weight and metastatic lesions (liver, lymph node and peritoneal) were reduced 3- to 5-fold by the combination therapy as compared with 1.5- to 2-fold with 3G4 and gemcitabine alone, respectively. Treatment of tumor bearing animals with the combination therapy increased macrophage infiltration into the tumor mass 10-fold and reduced microvessel density in the tumor by 2.5 fold compared with tumors from untreated animals. Gemcitabine alone and 3G4 alone were less effective than the combination of the 2 agents together. The additive therapeutic effect of both agents appears to be because chemotherapy increases PS exposure on tumor vascular endothelium and amplifies the target for attack by 3G4. In conclusion, 3G4 enhanced the anti-tumor and anti-metastatic activity of gemcitabine without contributing to toxicity. PMID- 16353143 TI - Enhancement of human papilloma virus type 16 E7 specific T cell responses by local invasive procedures in patients with (pre)malignant cervical neoplasia. AB - It has been suggested that local invasive procedures may alter the natural course of (pre)malignant cervical disease. This could be due to partial excision of the lesions, or via induction of cellular immunity against human papillomavirus (HPV) by the local invasive procedures. We studied the influence of local invasive procedures on HPV-16 E7 specific immune responses in patients with different grades of cervical intra-epithelial neoplasia (CIN) and different stages of cervical cancer. Blood was obtained at intake and after invasive procedures from patients with CIN or cervical cancer. Antigen specific T-cell responses were measured by IFN-gamma ELISPOT analysis, after stimulation with recombinant HPV-16 E7 protein. As expected, HPV-16 E7 specific IFN-gamma T cell responses were more frequent in HPV-16 DNA positive patients compared with that in HPV-16 DNA negative patients (39/50 vs. 16/36, (p=0.006, chi2 test). After invasive procedures, a small number of HPV-16 DNA positive CIN patients, but a considerable proportion of HPV-16 DNA positive cervical cancer patients, showed an enhancement of T cell responses against HPV-16 E7. Induction of T cell reactivity was most pronounced in cervical cancer patients who had undergone previous invasive procedures. Both CD4+ and CD8+ T cells showed E7 specific IFN gamma production upon in-vitro stimulation. Our study shows that invasive procedures may enhance HPV-specific cell-mediated immunity in a considerable number of patients with cervical cancer, but in only a minority of CIN patients. Our data indicate that invasive procedures should be considered as possible confounding factors when analyzing the effectiveness of therapeutic immunization studies, especially, when induction of HPV-specific immune responses is used as intermediate end-point. PMID- 16353144 TI - Lung cancer risk and occupational exposure to meat and live animals. AB - An increased risk of lung cancer has been reported for butchers and meat workers in several cohort studies, although confounding from tobacco smoking could not be ruled out in any of these studies. These exposures, as well as a potential risk associated with contact with live animals, are addressed here in a large case control study with full adjustment for smoking. More than 5,900 subjects were included in a case-control study conducted in 7 European countries. For each job they employed local experts who assessed the exposure to a number of occupational agents, including (i) meat aerosols and (ii) live animals, on the basis of detailed occupational questionnaires. Information on tobacco consumption and other risk factors was also collected. A small increased risk of lung cancer was observed with exposure to meat aerosols, after adjusting for smoking, (odds ratio (OR)=1.27, 95% confidence interval (CI): 0.92, 1.75), which was most apparent for the upper tertile of cumulative exposure (OR=1.73, 95% CI: 1.03, 2.92). A similar overall effect was observed for exposure to live animals, with an increased risk observed for a high frequency of exposure, (OR=1.69, 95% CI: 1.21, 2.36) and a high intensity of exposure, (OR=1.85, 95% CI: 1.16, 2.94), with significant trends for increasing frequency (p=0.012), intensity (p=0.015) and cumulative exposure (p=0.024). In conclusion, this study provides evidence for an association between exposure to meat aerosols and lung cancer apparent in the highest tertile of exposure. The authors identified a more consistent association with exposure to live animals. PMID- 16353145 TI - Celecoxib induces anoikis in human colon carcinoma cells associated with the deregulation of focal adhesions and nuclear translocation of p130Cas. AB - Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, is effective as chemopreventive against colon cancer and it is the only nonsteoroidal antiinflammatory drug approved by the FDA for adjuvant therapy in patients with familial adenomatous polyposis. It is also being evaluated, within Phase II and III clinical trials, in combination with standard chemotherapy to treat sporadic colorectal cancer. Nevertheless, its antitumor mechanism of action is still not fully understood. In this study, we have evaluated the in vitro growth inhibitory effect of celecoxib in colon carcinoma cells and analyzed its mechanism of action. We report that the deregulation of the focal adhesion assembly protein Crk-associated substrate 130 kDa (p130Cas) by celecoxib plays a relevant role in the cytotoxic effect of this drug. Thus, celecoxib induces the proteolysis of p130Cas and the nuclear translocation of the 31 kDa generated fragment leading to apoptosis. Furthermore, overexpression of wild-type p130Cas reverts, in part, the growth inhibitory effect of celecoxib. In contrast, FAK and AKT do not appear to be involved in this activity. Our data suggest, for the first time, that the antitumor mechanism of action of celecoxib includes the induction of anoikis, an effect that is not related to COX-2 inhibition. Besides providing new insights into the antitumor effect of celecoxib, this novel mechanism of action holds potential relevance in drug development. Indeed, our results open the possibility to develop new celecoxib derivatives that induce anoikis without COX-2 inhibition so as to avoid the cardiovascular toxicity recently described for the COX-2 inhibitors. PMID- 16353146 TI - Expression of multiple epigenetically regulated cancer/germline genes in nonsmall cell lung cancer. AB - Cancer/germline (CG) antigens represent promising targets for widely applicable mono- and multiantigen cancer vaccines for nonsmall cell lung cancer (NSCLC). Since little is known about their composite expression in this tumor type, we analyzed 7 CG genes (MAGE-A3, NY-ESO-1, LAGE-1, BRDT, HOM-TES-85, TPX-1 and LDHC) in 102 human NSCLC specimens. About 81% of NSCLC express at least 1 and half of the specimen at least 2 CG genes. Activation of most of these genes occurs more frequently in squamous cell cancer than in adenocarcinomas. Even though we found all genes but one to be regulated by genomic methylation, not all of them are co expressed. In particular, combining CG genes not localized on the X-chromosome may provide effective treatment for an extended number of patients. PMID- 16353148 TI - A functional polymorphism in the matrix metalloproteinase-1 gene promoter is associated with susceptibility and aggressiveness of head and neck cancer. AB - Matrix metalloproteinases (MMPs) play an important role in several steps of cancer development. A single guanine insertion polymorphism (2G) in the MMP1 promoter sequence at -1,607 creates an Ets binding site and thus results in enhancing transcriptional activity. This study aimed to evaluate the contribution of this 2G polymorphism on susceptibility and aggressiveness of HNSCC. A panel of HNSCC cell lines and peritumoral fibroblasts were examined for the MMP1 genotypes and expression levels. Genomic DNA was extracted from peripheral blood of 300 patients with newly diagnosed HNSCC and from 300 age- and gender-matched cancer free controls. Genotyping was carried out using a PCR-RFLP assay. The levels of MMP1 mRNA expression were evaluated by the quantitative RT-PCR and a correlation with different genotype was determined. Odds ratio (OR) for cancer risk were calculated using multivariate logistic regression. In addition, a correlation between the 2G/2G genotype and clinicopathological parameters was examined. Eleven out of 18 HNSCC cell lines showed the 2G/2G genotype (61%) and only 1 cell line had the 1G/1G genotype (5.6%). Cell lines with the 2G/2G genotype expressed significantly higher mean MMP1 mRNA level than those with other genotypes. In clinical model, subjects carrying the homozygous 2G/2G genotype had a higher risk of head and neck cancer compared with subjects with other genotypes (adjusted OR: 2.28; 95% CI: 1.58-3.27), controlling for major confounders. A correlation between promoter polymorphisms and the levels of MMP1 expression in cancer tissues was found, and this 2G/2G genotype was correlated with the adverse clinicopathological parameters. Finally, the highest level of MMP1 enhancement was demonstrated in the coculture of tumor cells and peritumoral fibroblasts of 2G homozygotes. These findings suggest that the presence of 2G polymorphism at the MMP1 promoter is associated with the development and progression of HNSCC. PMID- 16353147 TI - Inhibition of bone-derived insulin-like growth factors by a ligand-specific antibody suppresses the growth of human multiple myeloma in the human adult bone explanted in NOD/SCID mouse. AB - Multiple myeloma (MM) is a fatal disease that affects plasma cells. Patients with MM have 1 or more osteolytic lesions in their bone tissues, where insulin-like growth factors (IGFs; IGF-I and IGF-II) are mainly stored. The role of bone derived IGFs in the development of MM has not been extensively studied because reliable animal models are lacking. We established an animal model using a human MM cell line, RPMI8226, in nonobese diabetic/severe-combined immunodeficient (NOD/SCID) mice implanted with human adult bone (HAB) fragments. Treatment with an anti-human IGF-neutralizing monoclonal antibody, KM1468, inhibited the IGF-I stimulated phosphorylation of type-I IGF receptors (IGF-IR) in RPMI8226 cells and the activation of the downstream PI3-K/Akt signaling pathway in vitro. KM1468 inhibited IGF-I-mediated RPMI8226 cell growth in a dose-dependent manner. In the NOD/SCID-HAB model, treatment with KM1468 significantly inhibited the growth of RPMI8226 cells (p<0.02). These results indicated that the growth of MM cells was predominantly stimulated not by serum-derived IGFs, but by bone-derived IGFs. Furthermore, the targeting of bone-derived IGFs, using a neutralizing antibody, may offer a new therapeutic strategy for MM. PMID- 16353149 TI - Definition of an HPV18/45 cross-reactive human T-cell epitope after DNA immunisation of HLA-A2/KB transgenic mice. AB - Although human papillomavirus (HPV) types 16 and 18 are the most common types associated with cervical cancer worldwide, other related HPV types such as HPV 35, 45 and 58 have significant prevalence in geographically distinct populations. For development of global prophylactic and therapeutic vaccine strategies, it is important to study immune responses against these viruses and to define the degree of cross-reactivity between related HPV types. To investigate the potential for T cell cross-reactivity after vaccination, HLA-A2/Kb transgenic mice were immunised with DNA plasmid constructs containing HPV18 and 45 E6 and E7. Splenocytes from immunised mice were tested in direct ELIspot assays against overlapping pools of HPV 18 peptides. Immunisation with either HPV18 or HPV45 E6 DNA produced dominant T cell responses against an epitope (KCIDFYSRI) that was shared between HPV18 and HPV45. This peptide was shown to bind to HLA-A*0201 but not Db or Kb molecules on the cell surface. Furthermore this peptide was shown to be immunogenic in vitro to human T cells from 2 out of 3 HLA-A2+ healthy donors. Collectively, these results demonstrate that HPV 18 and 45 E6 DNA vaccines are immunogenic in mice and demonstrate that cross-reactive T cell responses against closely related HPV types can be induced in vivo. The use of the HLA-A2/Kb transgenic mice allowed definition of an HLA-A*0201 binding peptide epitope that would have been rejected on the basis of predicted major histocompatibility complex binding affinity. PMID- 16353151 TI - Body size and composition and the risk of gastric and oesophageal adenocarcinoma. AB - Although evidence has been mounting that obesity may be related to the increased incidence of oesophageal and gastric cardia malignancies, these reports (mainly case-control studies) have relied on imperfect measures of obesity such as body mass index (BMI), and generally have not clearly distinguished between anatomical subsites within the oesophagus and stomach. In a prospective study of people aged 27-75 years, we directly measured fat mass and fat-free mass (using bioelectrical impedance analysis), height, weight and waist and hip circumferences. Among 41,295 people followed on average for 11.3 years, 30 cases with cancers in the gastric cardia or lower third of the oesophagus and 68 cases with noncardia gastric adenocarcinomas were ascertained via the population cancer registry. The risk of adenocarcinoma of the lower oesophagus and gastric cardia was positively associated with BMI with a hazards ratio (HR) and (95% confidence interval) for people with BMI>or=30 kg/m2 compared with those<25 kg/m2, of 3.7 (1.1-12.4), an HR per 10 cm increase in waist circumference of 1.46 (1.05-2.04), and a HR per 10 kg increase on fat-free mass of 2.06 (1.15-3.69). Noncardia gastric adenocarcinoma showed little relationship with body size. We observed an increased risk of adenocarcinoma of the lower oesophagus and gastric cardia associated with increased BMI, central adiposity and the nonfat component of weight, but found no association with noncardia gastric adenocarcinoma. An increasing prevalence of obesity may be associated with the increasing incidence of gastro-oesophageal cancer observed in many populations. PMID- 16353152 TI - Increased incidence of invasive breast cancer after the introduction of service screening with mammography in Sweden. PMID- 16353153 TI - Enhancing effects of simultaneous treatment with sodium nitrite on 2-amino-3 methylimidazo[4,5-f]quinoline-induced rat liver, colon and Zymbal's gland carcinogenesis after initiation with diethylnitrosamine and 1,2 dimethylhydrazine. AB - Combined effects of sodium nitrite (NaNO2) and 2-amino-3-methylimidazo[4,5 f]quinoline (IQ) on liver, colon and Zymbal's gland carcinogenesis were assessed using a rat two-stage carcinogenesis model, with a focus on involvement of oxidative stress. Male 6-week-old F344 rats were given a single intraperitoneal injection of 200 mg/kg of diethylnitrosamine and 4 subcutaneous injections of 40 mg/kg of 1,2-dimethylhydrazine for initiation. Then, they were administered 0 or 300 ppm IQ in the diet or 0, 0.1 or 0.2% NaNO2 in their drinking water for 27 weeks. The treatment with NaNO2+IQ significantly enhanced colon and Zymbal's gland carcinogenesis and tended to enhance hepatocarcinogenesis. The incidence of lung tumors in the IQ-treated groups was significantly increased as compared with the initiation alone group. In a second experiment, male rats were given IQ or NaNO2 under the same conditions as before for 1 week, and at sacrifice, their liver and colon tissue or mucosa were collected for analysis of 8 hydroxydeoxyguanosine (8-OHdG), thiobarbituric acid reactive substances (TBARS), acrolein-modified protein and the bromodeoxyuridine-labeling index (BrdU-LI) (in the colon). In the colon, 8-OHdG, acrolein-modified protein levels and BrdU-LI were significantly increased by the combined treatment. These results indicate that the treatment with NaNO2 enhances IQ-induced colon and Zymbal's gland carcinogenesis in rats and that oxidative DNA damage and lipid peroxidation may partly be involved, especially in the colon. In addition, this experiment showed that IQ can act as a potent lung carcinogen in rats. PMID- 16353154 TI - Cytogenetic damage and genetic variants in the individuals susceptible to arsenic induced cancer through drinking water. AB - In West Bengal, India, more than 300,000 arsenic-exposed people are showing symptoms of arsenic toxicity, which include cancers of skin and different internal organs. Since only 15-20% of the exposed population manifest arsenic induced skin lesions, it is thought that genetic variation might play an important role in arsenic toxicity and carcinogenicity. A total of 422 unrelated arsenic-exposed subjects (244 skin-symptomatic and 178 asymptomatic) were recruited for this study. Cytogenetic damage, as measured by chromosomal aberrations in lymphocytes and micronuclei formation in oral mucosa cells, urothelial cells and binucleated lymphocytes, was studied in unexposed, skin symptomatic and asymptomatic individuals with similar socioeconomic status. Identification of null mutations in GSTT1 and GSTM1 genes were carried out by PCR amplification. GSTP1 SNPs, implicated in susceptibility to various cancers, were assessed by PCR-RFLP method. Symptomatic individuals had higher level of cytogenetic damage compared to asymptomatic individuals and asymptomatic individuals had significantly higher genotoxicity than unexposed individuals. No difference in allelic variants in GSTT1 and GSTP1 was observed between these 2 groups. Incidence of GSTM1 null gene frequencies was significantly higher in the asymptomatic group. Individuals with GSTM1-positive (at least one allele) had significantly higher risk of arsenic-induced skin lesions (odds ratio, 1.73; 95% confidence interval, 1.24-2.22). These results show a protective role of GSTM1 null in arsenic toxicity. This study also indicates that asymptomatic individuals are sub clinically affected and are also significantly susceptible to arsenic induced genotoxicity. PMID- 16353155 TI - Comparative genomic hybridization analysis on male breast cancer. AB - The spectrum of genetic alterations in primary male breast cancer is not well established. We analyzed chromosomal imbalances in 39 tumor samples from primary male breast cancer by comparative genomic hybridization (CGH) and correlated CGH findings with clinicopathological factors. Chromosomal gains were most frequent at 1q (46%), 8q (46%), 16p (36%), 17q (36%), Xq (28%), 20q (26%) and Xp (18%). Losses were most commonly observed at 8p (36%), 16q (28%), 13q (28%), 6q (18%), 11q (18%) and 22q (18%). Gains at 16p, 20q and Xq and losses at 13q correlated significantly with higher degree of cytogenetic complexity. Significant associations with clinicopathological factors were observed for +8q and -16q with larger tumor size and -16q with lower proliferative activity and lower grade of malignancy. A comparison with reported CGH data from female breast cancer showed a similar pattern of chromosomal imbalances, including +1q, -8p, +8q, -13q, +16p, -16q, +17q and +20q. Our results indicate that male breast cancer shares a common pattern of imbalances with female breast cancer, suggesting that similar genetic events may underlie the development and progression of male and female breast cancer. PMID- 16353156 TI - Development of multidrug resistance due to multiple factors including P glycoprotein overexpression under K-selection after MYC and HRAS oncogene activation. AB - Multistep tumorigenesis is a form of microevolution consisting of mutation and selection. To clarify the role of selection modalities in tumor development, we examined two alternative evolutionary conditions, r-selection in sparse culture, which allows cells to proliferate rapidly, and K-selection in confluent culture, in which overcrowding constrains cell proliferation. Using MYC- and EJ-RAS transformed rat embryo fibroblasts, we found that K-selected cells acquired and stably maintained multidrug resistance (MDR) to DOX, VCR, MTX and Ara-C. Then, we examined the involvement of a number of factors potentially causal of the development of MDR, that is, ploidy, Tp53 mutation, doubling time and the expression levels of genes related to drug resistance. Although ploidy status and Tp53 mutations did not correlate with MDR, we found that Abcb1/Mdr1, encoding P glycoprotein (Pgp), was significantly upregulated after K-selection. Cyclosporin A, a competitive inhibitor of Pgp, increased the intracellular accumulation of DOX and reduced the resistance to it. Indeed, the population of Pgp-transfected cells significantly expanded under K-, but not under r-selection. In addition to Pgp upregulation, altered expression of other genes such as Cda/cytidine deaminase and Slc29a1/equilibrative nucleoside transporter 1 and prolonged doubling times were associated with MDR. This system reproduces events associated with MDR in vivo and would be useful for analysis of MDR development. PMID- 16353157 TI - Phenoxodiol, an experimental anticancer drug, shows potent antiangiogenic properties in addition to its antitumour effects. AB - Phenoxodiol (2H-1-benzopyran-7-0,1, 3-[4-hydroxyphenyl], PXD) is a synthetic analogue of the naturally-occurring plant isoflavone and anticancer agent, genistein. PXD directly induces mitotic arrest and apoptosis in most cancer cells and is currently undergoing clinical trials, as a chemotherapeutic in ovarian and prostate cancers. We show here that PXD also exhibits potent antiangiogenic properties. Thus, it inhibited endothelial cell proliferation, migration and capillary tube formation and inhibited expression of the matrix metalloproteinase MMP-2, a major matrix degrading enzyme. Importantly, we demonstrate that PXD is functional in vivo since it inhibited the extent of capillary tube invasion in an in vivo model of angiogenesis. We show that phenoxodiol inhibits hallmarks of endothelial cell activation, namely TNF or IL-1 induced E-selectin and VCAM-1 expression and IL-8 secretion. However, PXD had no effect on unstimulated endothelial cells. We also describe that PXD inhibits the lipid kinase sphingosine kinase, which recently has been implicated in endothelial cell activation and angiogenesis as well as oncogenesis. Thus, our results suggest that PXD may be an effective anticancer drug targeting the two drivers of tumour growth--the proliferation of the tumour cells themselves and the angiogenic and inflammatory stimulation of the vasculature. PMID- 16353158 TI - Frequent EGFR mutations in noninvasive bronchioloalveolar carcinoma. AB - Mutations of the epidermal growth factor receptor gene (EGFR) have been reported to be present in a considerable fraction of lung adenocarcinomas showing dramatic response to EGFR tyrosine kinase inhibitors. To clarify pathogenic significance of the mutations for the development of lung adenocarcinoma, we investigated stage I lung adenocarcinomas for the mutations. First, 107 cases of macrodissected stage I adenocarcinomas were examined for mutations in exons 18-21 of the EGFR gene. EGFR mutations were detected in 36 of the 107 cases (34%). In particular, among the stage I cases, the mutations were detected in 17 of 42 small-sized adenocarcinomas (60%) at low surface quantities (<50 ng). Importantly, this approach can be employed to transfect cells throughout a three-dimensional scaffold. Tissue engineering scaffolds that are prefabricated into various shapes from a range of materials could potentially employ this strategy for numerous applications. PMID- 16353174 TI - Impairment of nitric oxide output of conducting airways in primary ciliary dyskinesia. AB - Nasal nitric oxide (NO) concentration is dramatically reduced in primary ciliary dyskinesia (PCD). The aims of this study were to apply a multiple-flow NO analysis to investigate whether NO output from the bronchial tree was affected in a similar way to nasal NO output, and to search for a relationship between flow independent exchange parameters and airflow limitation. Multiple flow rate analysis of exhaled NO, allowing the calculation of maximum airway wall flux and alveolar NO concentration, was performed in 17 PCD patients (median age, 25-75th percentiles: 13.5, 12.1-17.6) with documented ultrastructural cilia abnormalities and 28 healthy subjects (16.0, 11.0-21.0). Median maximum airway wall flux and median alveolar NO concentration were significantly reduced in PCD patients compared to healthy subjects: 16.0, 7.5-29.5, vs. 25.0, 15.0-32.5 nl/min (P<0.05) and 2.5, 1.6-3.3, vs. 5.0, 3.6-6.5 ppb (P<0.01), respectively. Significant correlations between maximum airway wall flux and airflow limitation were found, i.e., resistance of respiratory system (rho=0.74, P<0.005), forced expiratory volume in one second (FEV(1))/VC (rho= -0.61, P<0.05), FEV(1) (rho=-0.52, P< 0.05), mid expiratory flow between 25 and 75% of forced vital capacity (MEF(25 75)) (rho=-0.54, P<0.05), and maximal instantaneous expiratory flow at 50% of the vital capacity (MEF(50)) (rho=-0.55, P<0.05). In conclusion, the impairment of NO output is less pronounced in the lower than in the upper (nasal) respiratory tract in PCD. A decrease in maximal NO output from conducting airways is associated with limited airflow impairment. PMID- 16353175 TI - Plasticity of the motor cortex in Parkinson's disease patients on and off therapy. AB - We used the paired associative stimulation (PAS) technique to investigate associative plasticity of the sensorimotor cortex in 16 Parkinson's disease (PD) patients off and on therapy and in 10 age-matched controls. After PAS, motor evoked potential (MEP) amplitudes increased more and the cortical silent period showed a reduced prolongation in patients off therapy than in controls. These changes lasted for at least 30 minutes. In addition, MEP amplitudes increased in a less focal manner in patients off therapy than in controls. After patients received dopaminergic therapy, these abnormalities normalized. The abnormal responsiveness of sensorimotor cortex neurons to PAS in PD patients off therapy probably reflects disordered plasticity within the motor cortex. PMID- 16353176 TI - Alarming arbitrariness in EU prescription and reimbursement criteria for anti dementia drugs. PMID- 16353177 TI - Comparison of motor, cognitive, and behavioral features in progressive supranuclear palsy and Parkinson's disease. AB - Major clinical features and global measures were systematically evaluated and compared in progressive supranuclear palsy (PSP) and Parkinson's disease (PD). In addition to gaze palsy and early postural instability in PSP, absence of levodopa induced dyskinesia, frontalis muscle overactivity, primitive reflexes, visuospatial impairment, and substantial frontal behavioral disturbances differentiated almost all patients with this disorder from PD. For PSP, behavioral changes related to severity of general disability, thereby challenging previous models of relationships between behavior, motor, and cognitive disturbance for this disorder. PMID- 16353178 TI - Feasibility of shortened methacholine challenge in preschool children. AB - The aim of our study was to assess the feasibility and safety of a recently suggested tripling-dose methacholine (Mch) challenge in preschool children. Fifty seven children aged 3-6 years were studied. Mch challenge was carried out using a tidal breathing method, with concentrations of 0.22, 0.66, 2.0, 6.0, and 18.0 mg/ml, at 5-min intervals, given by a Pari Turbo Boy compressor and Pari LC Plus nebulizer, for 1 min only. Oxygen saturation (SaO(2)) was monitored during the challenge. The challenge was terminated if there was wheeze, SaO(2) below 91%, or persistent cough. This final Mch dose was considered the provocative concentration inducing audible wheeze (PCW). Nine healthy children, 17 with cough and 25 with wheeze, completed the study. Mean output from nebulizers (SD) in these 51 children was 0.30 (0.05) ml/min. Geometric means for PCW in these groups were 2.88, 2.58, and 1.28 mg/ml Mch, respectively. The wheezing children were significantly more hyperresponsive than the coughing children (P < 0.05). A tripling-dose Mch protocol is safe and practicable in children over 3 years of age. A further reduction in nebulized dose may be needed for a more discriminatory test. PMID- 16353179 TI - Expression of chymase-positive cells in gastric cancer and its correlation with the angiogenesis. AB - BACKGROUND AND OBJECTIVES: Chymase is expressed in mast cells and induces angiogenesis via activation of angiotensin II and matrix metalloproteinase-9. However, it has been unclear whether chymase is involved in the pathophysiology of angiogenesis in gastric cancer. To clarify the contribution of chymase to angiogenesis in gastric cancer, we assessed the relationship between chymase positive cells and tumor angiogenesis. METHODS: We evaluated chymase-positive cells and microvessels using anti-human chymase and anti-CD34 antibodies in 168 cases of gastric cancer, respectively. RESULTS: Chymase-positive cells in gastric tumor region were significantly higher than the cells in normal region. The number of chymase-positive cells in the undifferentiated type of gastric tumor region was significantly higher than the one in the differentiated type. Specimens from patients with advanced histological stages of disease had more chymase-positive cells than those with early-stage disease. There was a significant positive correlation between chymase-positive cells and microvessels in gastric cancer specimens. Postoperative survival curves revealed that patients with a high number of chymase-positive cells had a poor prognosis. CONCLUSIONS: These results suggest that accumulation of chymase-positive cells in gastric cancer may lead to an increase of tumor angiogenesis, and may contribute to tumor growth and progression. PMID- 16353180 TI - The epidermal growth factor receptor gene sequence is highly conserved in primary gastric cancers. AB - BACKGROUND AND OBJECTIVES: Recent studies have disclosed the presence of somatic mutations in the epidermal growth factor receptor (EGFR) gene in non-small cell lung cancers (NSCLC), and susceptibility to the EGFR tyrosine kinase inhibitor (gefitinib) was determined by the presence of mutations in the kinase domain of this gene. We thus predicted a clinical benefit of gefitinib against the 12% of primary colorectal cancers exhibiting a mutation reflective of this potential distinctive susceptibility. PATIENTS AND METHODS: The mutation status of the kinase domain in EGFR in different primary cancers has important clinical consequences, because the presence of a mutation is recognized as a reliable indicator for the effectiveness of gefitinib administration. In the current study, we investigated the presence of somatic mutations in exons 18-21 coding the ATP-binding domain in five gastric cancer cell lines and 39 primary gastric cancers and their corresponding normal tissues. RESULTS AND CONCLUSIONS: The kinase domain of EGFR is highly conserved in whole gastric cancer cell lines and cases, therefore treatment with gefitinib is unfortunately not recommended for such malignancy. PMID- 16353181 TI - Upregulation of thymidine phosphorylase in rectal cancer tissues by mitomycin C. AB - BACKGROUND AND OBJECTIVES: We have investigated the regulation by mitomycin C (MMC) of thymidine phosphorylase (dThdPase) and dihydropyrimidine dehydrogenase (DPD), which enhances or reduces the efficacy of capecitabine and its metabolite 5'-deoxy-5-fluorouridine (5'-DFUR), in rectal cancer tissues. MATERIALS AND METHODS: In 31 patients with a rectal cancer, tumor biopsies were performed before and after pre-operative venous administration of 4 mg/m2, 6 mg/m2, or 10 mg/m2 of MMC. The dThdPase and DPD levels in the biopsy and surgical specimens were measured using ELISA, and immunostaining for dThdPase was performed. RESULTS: The fitting multiple linear regression models indicated that the dThdPase levels increased after MMC administration, in particular in the patients with a pre-treatment dThdPase level less than 56.2 U/mg protein (median value). The time course analysis indicated that the increase in the dThdPase level by 4 mg/m2 of MMC administration continued for 3 weeks. The dThdPase/DPD ratio was increased after MMC administration in patients with a pre-treatment dThdPase/DPD ratio less than 1.79 (median value). MMC enhanced the expression of dThdPase protein both in the tumor cells and in the stromal cells. The disease free survival rate in the Dukes B or C patients with a high dThdPase/DPD ratio in surgical specimen who received 5'-DFUR based adjuvant chemotherapy tended to be higher than that in those with a low dThdPase/DPD ratio. CONCLUSION: MMC may upregulate the dThdPase level and the dThdPase/DPD ratio in rectal cancer tissues. Combined use of MMC with capecitabine or 5'-DFUR may offer a more effective colorectal cancer therapy. PMID- 16353182 TI - A new laparoscopic-assisted hepatectomy (LAH) utilizing radiofrequency ablation and high-frequency electrocautery. AB - Compared to tumors located at the edge of the liver, tumors located directly on the liver surface are often difficult to resect, especially when the organ is cirrhotic. To remove tumors at this location, our group has developed a round high-frequency electrosurgical knife that allows hemispherical resection of the liver tissue. This study describes a new laparoscopic-assisted hepatectomy utilizing radiofrequency ablation of the tissue surrounding the tumor and high frequency electrocautery. When compared with the laparoscopic hepatectomy, this new procedure seems to offer a safer, more effective, and less time-consuming means of resecting tumors on the liver surface. PMID- 16353183 TI - Major extremity amputation for nodal metastasis from squamous cell carcinoma. AB - The regional lymph node basin is usually the first site of spread in cutaneous squamous cell carcinoma (SCC). Regional metastases are usually treated with comprehensive nodal dissection, sometimes followed by radiation and/or chemotherapy. Occasionally, when disease recurs after these conventional therapies, major extremity amputation may be the only way to render patients free of disease. We present three cases for which major extremity amputation was necessary. One patient underwent forequarter amputation for nodal metastasis, which had invaded the brachial plexus. Another patient underwent hip disarticulation for femoral bony invasion from inguinal nodal metastasis. The third patient underwent a hemipelvectomy for nodal metastasis, which had eroded the femoral artery and pelvic bone. These cases show that major extremity amputation can be both palliative and curative in patients with SCC. PMID- 16353184 TI - The trans-metastasis hepatectomy (through metastases previously ablated with radiofrequency): results of a 13-case study of colorectal cancer. AB - BACKGROUND: Curative trans-metastasis liver resection immediately following radiofrequency (RF) destruction is a new technique that enables the proposition of a curative approach to patients with bilateral, unresectable liver metastases (LM), when the only possible future hepatectomy plane passes through a LM. Firstly, the ill-sited LM, which is located in the only feasible future hepatectomy plane, is ablated using RF; the hepatectomy is then performed through this ablated LM. AIM: The aim of this study is to report the feasibility and efficacy of this new approach, known as post-RF-trans-metastasis-hepatectomy (PRFTMH). MATERIALS AND METHODS: Thirteen patients with colorectal primary cancer were treated with PRFTMPH between January 2000 and May 2004. The mean number of LMs per patient was 10.7. Preoperative hypertrophy of the future remaining liver was achieved by selective portal vein embolization in eight patients. RESULTS: The mortality rate was 7.6% (one death), and morbidity was 24%. No local recurrence was observed at the PRTMPH site after a mean follow-up of 19.4 months (range: 47-10), demonstrating the efficacy of this technique. All patients, except those who died postoperatively, are currently alive; the median survival has not yet been attained after a mean follow-up of 19.4 months. CONCLUSION: TMPRFH is a new and safe technique, combining RF ablation and trans-RF hepatectomy, which makes it possible to propose a curative approach in certain patients with non-resectable bilateral LMs. PMID- 16353185 TI - Isolated tumor cells in lymph nodes are not a prognostic marker for patients with stage I and stage II colorectal cancer. AB - BACKGROUND AND OBJECTIVES: Lymph node metastasis is an important prognostic factor for colorectal cancer (CRC). The relationship between isolated tumor cells (ITC) in lymph nodes and the prognosis is controversial. The aim of this study is to evaluate the prognostic importance of ITC in lymph nodes from patients with stage I and II CRC. METHODS: We examined all the lymph nodes obtained during surgery from 121 consecutive patients with stage I and II CRC after a curative resection in 1999 by using monoclonal antibody against cytokeratin (MNF116). Clinicopathologic data and follow-up information were obtained from all patients. The mean follow up duration was 57 months. RESULTS: ITC were found in 50% (60/121) of the patients. The mean number of examined lymph node was 19.3 +/- 9.4. The presence of ITC was not related to clinicopathologic factors such as age, sex, location of tumor, tumor size, tumor depth, differentiation, lymphovascular invasion, and the preoperative CEA level, except for the tumor gross type (P = 0.002). There were no statistically significant differences for the recurrence rate (10/60 = 16.7% and 9/61 = 14.8%, respectively), and disease free survival (P = 0.809) between the ITC positive group and ITC negative group. CONCLUSIONS: The presence of ITC detected by immunohistochemical staining does not have an influence on the prognosis of patients with stage I and II CRC. PMID- 16353186 TI - Pseudomyxoma peritonei and the urinary tract: involvement and treatment related complications. AB - BACKGROUND: Pseudomyxoma peritonei (PMP) is a rare clinical syndrome characterized by intraperitoneal accumulation of mucus produced by neoplastic cells of mostly appendiceal origin. The aim of this study was to analyze primary and secondary involvement and treatment-related complications of the urinary tract in PMP. METHODS: A retrospective study of 92 patients with PMP, treated by cytoreduction and intraoperative hyperthermic intraperitoneal chemotherapy (HIPEC) at The Netherlands Cancer Institute between 1996 and 2004. RESULTS: Seven patients presented with involvement of the urinary tract. Major urologic complications occurred in five patients, of which two had secondary involvement of the urinary tract. Major urologic complications consisted predominantly of surgical complications related to the bladder. All patients with secondary involvement and/or urologic complications had undergone previous pelvic surgery. CONCLUSIONS: The urinary tract is rarely involved in patients with PMP. Secondary involvement is mostly observed and may be a result of seeding of PMP of pelvic origin after prior pelvic surgery. There is a low urologic complication risk of treatment with cytoreduction and HIPEC. The combination of secondary involvement and previous pelvic surgery is an omen of treatment-related urologic complications, necessitating (surgical) re-interventions and further management in close collaboration with urologists. PMID- 16353188 TI - Split therapy: planned neck dissection followed by definitive radiotherapy for a T1, T2 pharyngolaryngeal primary cancer with operable N2, N3 nodal metastases--a prospective study. AB - BACKGROUND: The management of patients with a small pharyngolaryngeal cancer (T1 and T2) with large nodal metastases is a subject of debate. We present data on the feasibility and outcome of treating these patients with surgery for the nodal metastases followed by definitive radiotherapy. METHODS: Prospective study of 59 patients of small pharyngolaryngeal primary squamous carcinomas with operable (N2/N3) nodal metastasis treated with neck dissection followed by radiotherapy. RESULTS: Complete nodal clearance was achieved in 54 (90%). The mean nodal size was 4 cm and extranodal extension was seen in 88% of patients in the study group. There were no significant postoperative complications. Median interval between surgery and radiotherapy was 23 days. Forty-nine patients (83%) started their RT within 6 weeks of surgery. With a median follow-up of 25 months, the disease free and overall survival was 54% and 60% (5 years). CONCLUSION: The management of patients with a radiocurable pharyngolaryngeal primary with large nodes by this approach is a feasible option with adequate control and survival. PMID- 16353189 TI - Extended right hepatectomy for hilar cholangiocarcinoma with resection of the left hepatic duct prior to hepatic resection. AB - Extended hepatectomy for hilar cholangiocarcinoma results in high operative or in hospital mortality rates despite of the recent progress in perioperative management. As a new procedure to prevent postoperative hepatic failure in hilar cholangiocarcinoma infiltrating predominantly the right hepatic duct, we devised a combination of extended right lobectomy plus caudal lobectomy with resection of the left hepatic duct prior to hepatic resection by utilizing intraoperative cholangiography, and applied the procedure to a 70-year-old patient. He had a favorable postoperative course and remains recurrence-free at 4 years after operation. This is a procedure for confining the extent of hepatectomy to the minimum necessary, aiming at curative resection of hilar cholangiocarcinoma. PMID- 16353190 TI - Increased expression of matrix metalloproteinase-2 and 9 and human papilloma virus infection are associated with malignant transformation of sinonasal inverted papilloma. AB - BACKGROUND AND OBJECTIVES: In this study, we identified matrix metalloproteinase (MMP)-2 and 9 expression in exophytic papilloma (EP), inverted papilloma (IP) with dysplasia, IP with carcinoma, and invasive squamous cell carcinoma (SCC). We also compared the presence of MMP-2 and 9 with human papilloma virus (HPV) infection. METHODS: The nasal tissue were stained with monoclonal antibodies to MMP-2 and 9. The results were analyzed using quantitative immunohistochemical analysis. In situ hybridization studies for HPV DNA for 6/11, 16/18, and 31/33 were also performed on the tissue. RESULTS: Significant increase of MMP-2 and 9 were observed in IP with moderate and severe dysplasia, IP with carcinoma, and invasive SCC compared to control nasal mucosa. Among IP, HPV 6/11-positive was present in 41% and HPV 16/18-positive was present in 31%. In HPV 6/11 and 16/18 positive IP, significant increase of MMP-2 and 9 were observed. CONCLUSIONS: Precancerous lesions of IP exhibited elevated levels of MMP-2 and 9 and these expression may be associated with early events in IP carcinogenesis. HPV infection would be an early event in a multistep process of malignant formation of IP. PMID- 16353192 TI - Repeat hepatectomy for recurrent colorectal liver metastases. AB - PURPOSE: Liver resection represents the best and potentially curative treatment for metastatic colorectal cancer (MCC) to the liver. After resection, however, most patients develop recurrent disease, often isolated to the liver. The aim of this study was to determine the value of repeat liver resection for recurrent MCC and to analyze the factors that can predict survival. PATIENTS AND METHODS: From January 1992 to October 2002, 42 patients from a group of 168 patients resected for MCC were submitted to 55 repeat hepatectomies (42 second, 11 third, and 2 fourth hepatectomies). Records were retrospectively reviewed. The primary tumor was carcinoma of the colon in 26 patients and carcinoma of the rectum in 16 patients. Liver metastases were synchronous in 24 patients (57.1%). RESULTS: There were 25 men and 17 women with the mean age of 63.5 years (range: 34-80). There was no intraoperative or postoperative mortality. The morbidity rates were 9.5%, 14.3%, and 18.2% (P = 0.6) respectively after a first, second, or third hepatectomies. No patients needed reoperation. Operative duration was longer after a second or third hepatectomie than after a first hepatectomie without difference for operative bleeding. Overall 5-year survivals were 33%, 21%, and 36% respectively after a first, second, or third hepatectomies. Factors of prognostic value on univariate analysis included serum carcinoembryonic antigen levels (P = 0.01) during the first hepatectomy, the presence of extrahepatic disease (P = 0.05) and tumor size larger than 5 cm (P = 0.04) during the second hepatectomie. CONCLUSIONS: Repeat hepatectomies can provide long-term survival rates similar to those of first hepatectomies. PMID- 16353193 TI - Surgical technique for totally implantable access ports (TIAP) needs improvement: a multivariate analysis of 400 patients. AB - BACKGROUND: The objective of this study was to analyze factors that may have an impact on the failure rate of the surgical implantation technique for totally implantable access ports (TIAP) and to assess whether morbidity rates differ between a primarily successful surgical and a secondary Seldinger approach. METHODS: Four hundred consecutive patients receiving a primary TIAP in local anesthesia were included into this retrospective cohort study. A logistic regression-model was used to evaluate reasons for failure of the TIAP method. RESULTS: Three hundred eighteen (79.5%) patients had a successful TIAP procedure, 82 patients were intraoperatively converted to a Seldinger technique. Reasons for failure were: correct positioning impossible n = 54, no or only an undersized vessel for insertion available n = 17, other reasons n = 11. Logistic regression analysis did not reveal any significant factor for failure of the primary surgical approach. In the group with primarily successful TIAP, 8 of 318 patients (3%) developed complications compared to 7 of 82 patients (9%) converted to a Seldinger technique. CONCLUSION: Failure of the surgical approach for implanting totally implantable access ports is related to insertion and positioning. Conversion to a Seldinger technique results in a higher complication rate. A modified approach for surgical port placement should be considered in order to reduce complications. PMID- 16353194 TI - Differential expression of lysophosphatidic acid receptor-2 in intestinal and diffuse type gastric cancer. AB - BACKGROUND AND OBJECTIVES: Lysophosphatidic acid (LPA), a natural phospholipid, can modulate diverse cellular responses through LPA receptor, LPA1-4. Although LPA1 is known to be widely expressed in human tissues, the distribution of other LPA receptors is not characterized in malignant tissues. Recently, it was reported that malignant transformation resulted in aberrant expression of LPA2 in a various type of cancer, suggesting the positive role of LPA2 in tumor development. METHODS: We investigated the expression of the LPA2 receptor immunohistochemically in 204 gastric cancers and analyzed the relationship between the expression of LPA2 and clinicopathological features. RESULTS: LPA2 was preferentially expressed (67%) in intestinal-type cancer that was significantly higher than that in diffuse-type cancer (32%, P < 0.0001). The expression of LPA2 showed correlation with a higher rate of lymphatic and venous invasion, lymphatic metastasis, and resultingly tumor stage in diffuse-type cancer, but not in intestinal-type cancer. CONCLUSIONS: Our results highlight the possibility that LPA2 expression is an important process in the carcinogenesis of gastric cancer, especially in intestinal-type cancer. Since LPA can transactivate HGF receptor (c-Met) as well as EGF-receptor, LPA may promote the progression of gastric cancer in diffuse-type with high expression of c-Met. The development of LPA2-specific antagonists might have future therapeutic relevance in the treatment as well as prevention of gastric cancer. PMID- 16353195 TI - Expression of telomeric repeat binding factor 1 in non-small cell lung cancer. AB - BACKGROUND AND OBJECTIVES: Telomeric repeat binding factor 1 (TRF1) is crucial for forming and maintaining the protective telomeric structure. However, the relationship between TRF1 and non-small cell lung cancer(NSCLC) is not well understood. With this background, we investigated the expressions of the mRNA encoded by the TRF1 gene in cancer tissue and the paired non-cancerous tissue. We also examined whether TRF1 expression is correlated with histopathological features. METHODS: From October 2004 to August 2005, 40 patients with NSCLCs had undergone curative operations, including 29 males and 11 females. There were 20 cases of squamous cell carcinoma and 20 cases of adenocarcinoma. We measured the expression of TRF1 mRNA using RT-PCR on 40 surgically resected specimens and the paired non-cancerous tissues. RESULTS: TRF1 mRNA was significantly downregulated in cancer tissue compared with the paired non-cancerous tissue. Additionally, the expression of TRF1 mRNA was significantly associated with the grade of tumor differentiation. No significant difference of TRF1 mRNA level was found between sexes, or among different T-status, clinical stages, pathological subtypes, and lymph node metastasis. CONCLUSIONS: Downregulation of TRF1 mRNA expression appeared in lung cancer tissue. TRF1 may play a significant role in cell differentiation in NSCLC. PMID- 16353197 TI - Influence of substrate concentration on the stability and yield of continuous biohydrogen production. AB - The effect of substrate concentration (sucrose) on the stability and yield of a continuous fermentative process producing hydrogen was studied. High substrate concentrations are attractive from an energy standpoint as they would minimise the energy required for heating. The reactor was a CSTR; temperature was maintained at 35 degrees C; pH was controlled between 5.2 and 5.3, and the hydraulic retention time (HRT) was 12 h. Online measurements were taken for ORP, pH, temperature, %CO2, gas output and %H2, and data logged using a MatLAB data acquisition toolbox. Steady-state operation was obtained at 10, 20 and 40 g/L of sucrose in the influent, but a subsequent step change to 50 g/L was unsustainable. The hydrogen content ranged between 50% and 60%. The yield of hydrogen decreased as the substrate concentration increased from 1.7 +/- 0.2 mol/mol hexose added at 10 g/L, to 0.8 +/- 0.1 mol/mol at 50 g/L. Sparging with nitrogen improved the hydrogen yield by at least 35% at 40 g/L and at least 33% at 50 g/L sucrose. Sparging also enabled steady-state operation at 50 g/L sucrose. Addition of an extra 4 g/L of n-butyric acid to the reactor operating at 40 g/L sucrose increased the butyrate concentration from 9,830 to 18,900 mg/L, immediately stopping gas production and initiating the production of propionate, whilst the addition of 2 g/L taking the butyrate concentration to 12,200 mg/L did not do so. It was shown that operation at 50 g/L sucrose in a CSTR in butyrate fermentation is possible. PMID- 16353198 TI - Charles Manning Child (1869-1954): the past, present, and future of metabolic signaling. AB - Charles Manning Child's work focused on metabolic gradients and their influence on organismal development. Early in the 20th century, his work had considerable currency, but by the second half of the century he had become little more than a historical footnote. Yet today Child's ideas are once again topical. While there were issues of cause and effect that Child and his students were never able to address adequately, in hindsight the extent of his eclipse hardly seems warranted. In fact, the demise of Child's theories may have resulted from larger changes in the nature of biology in the early 20th century. Child frequently studied planarians, hydroids, and other animals that are capable of asexual, agametic reproduction, and his theories most clearly apply to such organisms. In contrast, Thomas Hunt Morgan, initially one of Child's competitors in studies of regeneration, later developed the field of transmission genetics based on fruit flies, which can only reproduce via gametes. Child's theories and model systems were largely casualties of the success of Morgan's mechanistic paradigm. Nevertheless, in modern biology metabolic gradients, recast in terms of redox signaling, have become central to understanding both normal and pathological development. PMID- 16353199 TI - Calcium plus vitamin D alters preneoplastic features of colorectal adenomas and rectal mucosa. AB - BACKGROUND: Calcium and vitamin D are chemopreventive agents for colorectal neoplasia. Studies of the effects of calcium and vitamin D on early surrogate markers of reduced risk, such as proliferation, have been limited to evaluation of the flat colorectal mucosa. Biologic changes that may occur in colorectal adenomas after chemopreventive regimens have not been reported. METHODS: In the current study, adenomatous polyps were transected, approximately 50% were removed for histologic examination, and the remnants tattooed before the administration of either calcium carbonate (1500 mg 3 times daily) plus vitamin D(3) 400 IU or a placebo for 6 months. At study end, polyp remnants were resected completely and were used for histologic examination. Immunohistochemical staining was performed in both flat mucosa and in polyp tissue. Proliferation was assessed by MIB-1 staining; apoptosis was assessed by terminal deoxyuridine triphosphate-biotin nick-end labeling, BAK, and Bcl-2 staining; and cytokeratin AE1, vitamin D receptor, MUC5AC mucin, and galectin-3 were assessed by immunohistochemistry. RESULTS: Nineteen patients, including 11 patients in the treatment group and 8 patients in the control group, completed the study. Proliferative indices fell both in flat mucosa and in polyps in the treatment group, and there were no significant changes in the control group. Apoptosis and Bcl-2 immunostaining were unchanged in both groups, but the frequency of BAK-immunostained cells in the interior of polyps rose significantly. Vitamin D receptor staining increased slightly and significantly in flat rectal tissue in the treatment group. There were no significant changes in galectin-3 staining, but a striking reduction in MUC5AC mucin staining in polyps was observed after treatment with calcium plus vitamin D. CONCLUSIONS: The administration of a calcium plus vitamin D chemopreventive regimen resulted in several changes in adenomatous tissue that may have contributed to reduced polyp formation. PMID- 16353200 TI - Domain interactions direct misfolding and amyloid formation of yeast phosphoglycerate kinase. AB - There are proteins that are built of two structural domains and are deposited full-length in amyloid plaques formed in various diseases. In spite of the known differences in the mechanisms of folding of single- and multidomain proteins, no published studies can be found that address the role of the domain-domain interactions during misfolding and amyloid formation. By the discovery of the role of domain-domain interactions, here we provide important insight in the submolecular mechanism of amyloid formation. A model system based on yeast phosphoglycerate kinase was designed. This system includes the wild-type yeast phosphoglycerate kinase and single-tryptophan mutants of the individual N and C terminal domains and the complete protein. Electron microscopic measurements proved that amyloid fibrils grow from all mutants under identical conditions as for the wild-type protein. Misfolding and amyloid formation was followed in stopped-flow and manual mixing experiments on the 1 ms to 4 days timescale. Tryptophan fluorescence was used for selective detection of conformational changes accompanying the formation of the amyloidogenic intermediates and the growth of amyloid fibrils. The interactions between the polypeptide chains of the two domains direct the misfolding process from the early steps to the amyloid formation, and influence the final structure. The kinetics of misfolding is different for the individual domains, pointing to the significance of the amino acid sequence. Misfolding of the domains within the complete protein is synchronized indicating that domain-domain interactions direct the misfolding and amyloid formation mechanism. PMID- 16353201 TI - A retrospective comparison of three sequential groups of patients with Recurrent/Refractory chronic lymphocytic leukemia treated with fludarabine-based regimens. AB - BACKGROUND: Combining therapeutics with single-agent activity has improved treatment for patients with many malignancies. Debate continues about the impact of treatment on survival in patients with chronic lymphocytic leukemia (CLL). Purine analogues are the most active agents for treatment of patients with CLL. Recently, it was shown that a chemoimmunotherapy regimen combining fludarabine (F), cyclophosphamide (C), and rituximab (R) (FCR) was very effective in treating patients with recurrent and/or refractory CLL. The objective of the current analysis was to determine whether improvements in treatment have had an impact on survival for patients with CLL. METHODS: Three nonoverlapping, sequential groups of patients enrolled on Phase II studies who received treatment with F (n = 251 patients), FC (n = 111 patients), or FCR (n = 143 patients) were analyzed. Pretreatment characteristics, responses to treatment, and overall survival were compared. RESULTS: Patients who were treated with FCR had a higher complete remission rate compared with patients who were treated with combined F and C or with F alone. Statistically significantly longer estimated median survival was noted for patients who received FCR. A Cox proportional hazards, multivariable model for overall survival that included all patients (n = 505) showed that patients who received FCR had longer survival (P < 0.0001) after adjusting for other significant (P < 0.05) pretreatment characteristics, including age, hemoglobin, beta-2 microglobulin, and the number of prior treatments. CONCLUSIONS: The results of this retrospective comparison of patients with recurrent and refractory CLL indicated a higher complete remission rate and the longest estimated survival for patients who were treated with FCR, providing the basis for randomized clinical trials of this regimen. PMID- 16353202 TI - The preoperative erythrocyte sedimentation rate is an independent prognostic factor in renal cell carcinoma. AB - BACKGROUND: Prognostic nomograms are used increasingly in clinical trials and to guide surveillance for patients with renal cell carcinoma (RCC). An elevated erythrocyte sedimentation rate (ESR) reportedly has been associated with a poor prognosis among patients with RCC, but the ESR is not incorporated into existing nomograms. Hence, the current study was conducted to expand on prior observations pertaining to the ESR as a prognostic indicator in patients with RCC. METHODS: The authors identified 3008 patients who underwent nephrectomy for RCC between 1970 and 2002. Disease-specific survival was estimated using the Kaplan-Meier method, and its association with the ESR and other clinical and pathologic features was evaluated using Cox proportional hazards regression analysis. RESULTS: A preoperative ESR was available for 1075 patients (35.7%), 501 of whom (46.6%) exhibited an elevated ESR, including 437 of 881 patients (49.2%) with clear cell RCC, 41 of 134 patients (30.6%) with papillary RCC, and 20 of 48 patients (41.7%) with chromophobe RCC. An elevated ESR was associated with adverse clinical, laboratory, and pathologic profiles for all three histologic subtypes. The risk ratios (RRs) and 95% confidence intervals (95% CIs) for death because of clear cell RCC, papillary RCC, and chromophobe RCC for patients with an elevated ESR were 3.6 (95% CI, 1.1-1.9), 3.8 (95% CI, 1.4-10.6), and 10.3 (95% CI, 1.2-89.5), respectively. The association between an elevated ESR and death from clear cell RCC persisted even after multivariate analysis (RR of 1.5; 95% CI, 1.2-2.0). CONCLUSIONS: An elevated ESR in patients with RCC suggested the presence of aggressive disease and poorer outcomes after surgical treatment. For patients with clear cell RCC, the ESR provided useful information above and beyond traditional prognostic algorithms, and it may be valuable for preoperative prognostication. PMID- 16353203 TI - Functional outcome after low-grade astrocytoma treatment in childhood. AB - BACKGROUND: The relatively high survival rate of patients with low-grade astrocytoma necessitates increasing attention to physical and psychosocial outcomes. The objective of the current study was to investigate functional outcomes among children who were treated for low-grade or pilocytic astrocytoma in different areas of the brain. METHODS: Functional outcomes were evaluated in the following domains: impairments, disabilities, handicaps, and quality of life (QOL). In a consecutive series, 38 children were included. Follow-up ranged from 3 years and 7 months to 11 years and 4 months after diagnosis. RESULTS: Approximately 61% of children had impairments and 10% had a severe disability. Handicaps were found in the domains of relationships, school, and behavior. Children who were treated for supratentorial tumors required significantly more special education, and children who were treated for infratentorial tumors had significantly more behavioral and social problems. QOL was decreased significantly in all domains except emotions. Children who had a diagnosis in adolescence reported a lower QOL in social functioning compared with younger children. Data analysis revealed that some deficits suddenly became apparent years after diagnosis. CONCLUSIONS: At long-term follow-up, children who had low grade or pilocytic astrocytomas were found to have poor functional outcomes, depending on tumor site, age, and recurrence. Children without deficits may develop severe cognitive, social, and behavioral deficits years after diagnosis, because of the phenomenon of "growing into deficit." Therefore, the authors suggest a long-term follow-up of children who are treated for low-grade or pilocytic astrocytomas at a young age to detect and subsequently offer support focused on the medical and cognitive impairments as well as on the behavioral and social consequences of their disease. PMID- 16353204 TI - Metastatic osteosarcoma. AB - BACKGROUND: The outcome of patients with metastatic osteosarcoma treated in two consecutive trials from 1986 to 1997 was analyzed to evaluate the efficacy of carboplatin-based multiagent chemotherapy and to identify prognostic factors. The initial study (OS-86) used ifosfamide, cisplatin, doxorubicin, and high-dose methotrexate, and the subsequent study (OS-91) used the same agents at similar doses, but carboplatin was substituted for cisplatin. METHODS: Twelve patients (median age, 15.1 yrs) were treated in OS-86 for osteosarcoma metastatic to the lung only (11 patients) or bone only (1 patient), and 17 patients (median age, 15.1 yrs) were treated in OS-91 for osteosarcoma metastatic to the lung only (12 patients), bone only (2 patients), lung and bone (2 patients), or other site (1 patient). RESULTS: Patients with metastatic disease enrolled in OS-86 and those with metastatic disease enrolled in OS-91 did not differ in terms of demographic features, histologic subtype, site of primary tumor, or site of metastases. There was a difference in survival according to treatment protocol (P = 0.054). All survivors (four of whom were enrolled in OS-86 and one of whom was enrolled in OS 91) had lung metastases only. Five-year survival estimates for patients with lung metastases only were 45.5 +/- 13.7% (OS-86) and 8.3 +/- 5.6% (OS-91) (P = 0.084). Unilateral lung metastases (P = 0.006), no more than three lung nodules (P = 0.014), and surgical remission (P = 0.001) were associated with improved survival probability. CONCLUSIONS: The poor outcome of patients with metastatic osteosarcoma treated in OS-91 justifies the use of cisplatin with its associated toxicity in patients with high-risk disease. PMID- 16353205 TI - Classic Kaposi sarcoma. Which KSHV-seropositive individuals are at risk? AB - BACKGROUND: Classic Kaposi sarcoma (CKS) is a relatively rare vascular disease primarily affecting human immunodeficiency virus (HIV)-uninfected elderly men. The infection with Kaposi sarcoma-associated herpesvirus (KSHV) is necessary for the establishment of Kaposi sarcoma (KS), although it is not sufficient. Thus, only a small fraction of KSHV-infected individuals develops KS. The cofactors that influence risk of KS among HIV-uninfected individuals are yet to be determined. The objective of the current study was to assess potential risk factors for CKS in the KSHV-infected Jewish population in Israel. METHODS: A case control study involved 35 CKS cases and 48 matched KSHV-infected controls. Lifestyle and medical history data from case patients and controls were compared by logistic regression analysis. RESULTS: In a multivariate analysis, the authors identified an age-related small increased risk for CKS in subjects originating from Asia and Africa. The risk for CKS increased, although not significantly statistically, in subjects who reported alcohol consumption, diabetes mellitus, herpes simplex, and asthma. No relation was found with cigarette smoking, family size, number of lifetime sexual partners, or sexually transmitted disease. CONCLUSIONS: A borderline increase in CKS risk among elderly subjects originating from Africa or Asia was identified. These results need to be further evaluated by larger studies. The authors believe that genetic and immunologic parameters may alter risk for CKS and, therefore, should also be investigated. PMID- 16353206 TI - Clinical trial accrual among new cancer patients at a community-based cancer center. AB - BACKGROUND: To the authors' knowledge, only limited data are available regarding clinical trial accrual patterns and the barriers encountered among newly diagnosed patients seen at community-based cancer centers. METHODS: In the current study, the authors prospectively collected clinical and sociodemographic data from all adult patients seen at a community-based cancer center who had new cancers diagnosed between 2003-2004. Clinical trial enrollment decisions were noted and factors that prevented accrual were identified. RESULTS: There was a total of 1012 new cancer patients. In 587 patients (58%), clinical trials appropriate for the diagnosis and stage of disease were not available. Among those patients for whom trials were available, 19.8% did not meet eligibility criteria, and only 9.9% of patients were enrolled. Although more trials were found to be available for women compared with men (51% vs. 32%; P < 0.01), the accrual rates were equal (11.2% vs. 7.6%; P = 0.24). Elderly patients comprised approximately 59.4% of those patients with available trials, but they were less likely to be enrolled (5.1% vs. 16.8%; P < 0.01). The major barriers to nonparticipation can be grouped into protocol limitations (68.1%), physician triage (16%), and patient decisions (15.9%). The overall accrual rate when all patients were included was 4% (42 of 1012 patients). CONCLUSIONS: At the study institution, participation in clinical trials is reported to be low. The unavailability of appropriate clinical trials represents the most significant barrier. Continuing efforts to encourage physicians and to educate patients remain necessary. If the current study findings are found to be applicable to other community-based cancer centers, making a larger variety of clinical trials available to the community may help to improve the accrual of patients to national cancer clinical trials. PMID- 16353207 TI - American founder mutation for Lynch syndrome. Prevalence estimates and implications. AB - BACKGROUND: Recently, a new founder mutation, an exon 1-6 deletion in a mismatch repair gene (MMR), MSH2, in nine kindreds with Lynch syndrome was reported. In 3 of the kindreds this mutation was traced by genealogy through 11-12 generations to a common founder, and thus termed the American Founder Mutation (AFM). Since then, 13 additional 'unrelated' kindreds with AFM were detected by a recently designed single polymerase chain reaction. This test might serve as first-line screening for Lynch syndrome mutations, provided AFM was prevalent, which is assessed in the current study. METHODS: The number of current AFM carriers and the incidence of Lynch syndrome caused by AFM were estimated based on population growth of mutation carriers derived from genealogy data. For cross-checking, its annual incidence was also estimated based on published epidemiology data. RESULTS: There are 18,981 (5th and 95th percentiles, 6038 and 34,466, respectively) expected current AFM carriers, or 160 (range 51-290) Lynch syndrome cases diagnosed per year due to AFM estimated based on genealogy data. The incidence estimate closely overlaps with that based on published epidemiology data, which is 114-400 cases per year. CONCLUSIONS: A large number of AFM carriers are likely to exist in the U.S., which harbors significant implications for cancer control. Given the ease of detection, testing for AFM not only among members of the existing AFM families, but also among all patients with Lynch syndrome in the U.S. is proposed. PMID- 16353208 TI - Efficacy of caspofungin against invasive Candida or invasive Aspergillus infections in neutropenic patients. AB - BACKGROUND: Neutropenia is an indicator of poor prognosis in patients with fungal infections. All available clinical trial experience from the caspofungin development program was reviewed to ascertain the efficacy of caspofungin in neutropenic patients with documented invasive aspergillosis (IA) or invasive candidiasis (IC). METHODS: The review was limited to neutropenic patients with proven IC or proven/probable IA at caspofungin onset. Data were available from four clinical trials. All patients had an absolute neutrophil count < 500/mm(3) at the initiation of caspofungin. In all cases caspofungin was administered as monotherapy at a dose of 50 mg/day, after a 70-mg loading dose. In all patients efficacy was assessed at the completion of caspofungin therapy. Success included complete and partial responses. RESULTS: Sixty-eight neutropenic patients were identified with documented invasive infection, including 27 with IC and 41 with IA. Most patients had acute or chronic leukemia. A favorable response was noted in 63% (17 of 27 patients) of patients with IC, including a 58% (14 of 24 patients) response as first-line therapy and a 100% (3 of 3 patients) response as salvage therapy. Success in candidemia was 68% (17 of 25 patients). Outcomes across the different Candida species were similar. Favorable responses were noted in 39% (16 of 41 patients) of patients with IA, including a 42% (5 of 12 patients) response as first-line therapy and 38% (11 of 29 patients) response as salvage therapy. Success by site of IA was 40% for pulmonary (12 of 30 patients), 43% for sinus (3 of 7 patients), and 25% for skin/disseminated site (1 of 4 patients). CONCLUSIONS: A review of the caspofungin database demonstrates that this echinocandin is effective in neutropenic patients with documented cases of IC or IA. PMID- 16353209 TI - Expression of the Bax inhibitor-1 gene in pulmonary adenocarcinoma. AB - BACKGROUND: The regulation of programmed cell death, or apoptosis, is crucial for normal development and for the maintenance of homeostasis. It has been shown that the novel antiapoptotic protein Bax inhibitor-1 (BI-1) represents a new type of regulator of cell death pathways controlled by Bcl-2 and Bax. METHODS: Surgically resected lung specimens were obtained from 32 patients with peripheral adenocarcinomas, and BI-1 gene expression was examined and compared with expression of the p53, bcl-2 and Bax genes. RESULTS: Fourteen of 32 tumors (43.8%) were positive for BI-1 gene expression by in situ hybridization. BI-1 gene expression in tumor specimens was significantly higher in adenocarcinomas with bronchioloalveolar carcinoma (BAC) and in adenocarcinomas of mixed subtypes with bronchioloalveolar spreading (14 of 17 tumors; 82.4%) than in carcinomas without it spreading. Patients who had BI-1-positive adenocarcinoma showed a relatively favorable prognosis compared with patients who had BI-1-negative adenocarcinoma. Eleven of 32 tumors (34.4%) were positive for the p53 protein, only 1 of 32 tumors (3.1%) was positive for the Bcl-2 protein, and 26 of 32 tumors (81.3%) were positive for the Bax protein. Protein expressions of p53, Bcl 2, and Bax, as detected by immunohistochemistry, were not associated with BI-1 gene expression. CONCLUSIONS: BI-1 gene expression was restricted to tumor cells with lepidic growth and was a prognostic factor for peripheral-type adenocarcinoma. It is believed that BI-1 gene expression is conserved evolutionarily and may act as a key regulator of the apoptotic pathway in BAC. PMID- 16353210 TI - Histologic subtypes as determinants of outcome in esophageal carcinoma patients with pathologic complete response after preoperative chemoradiotherapy. AB - BACKGROUND: The current study tested the hypothesis that the clinical outcome of patients with localized esophageal carcinoma after preoperative chemoradiotherapy (CTRT) depends on histology. METHODS: The authors stratified patients by adenocarcinoma (ACA) or squamous cell carcinoma (SCC) and compared the overall survival (OS) and patterns of failure among patients achieving pathologic complete response (pathCR) and or=0.05). In the ACA group, a greater portion of 0.2 for all analyses). However, the 95% confidence intervals were wide and included the possibility that even a minor delay in surgery might have a large impact on the probability of BCR. CONCLUSIONS: The time between biopsy and surgery does not appear to have a large effect on the risk of disease recurrence. Counseling patients on the importance of avoiding undue delay to surgery must be based on clinical judgment, particularly with respect to modifying advice based on the patient's risk. PMID- 16353214 TI - Flow cytometric comparison of RNA content in peripheral blood mononuclear cells of Down syndrome patients and control individuals. AB - OBJECTIVE: Trisomy 21 or Down syndrome (DS) is the most common genetic cause of mental retardation associated with the immunologic and other known defects. Extra chromosome 21 of DS patients contains an average of 40 extra copies of rRNA genes and the in vivo regulation of these genes' activity is not known. Because over 80% of total cellular RNA is rRNA, the measurement of total cellular RNA provides information on rRNA content. The aim of this work was to determine whether or not the additional chromosome 21 causes any increase in total cellular RNA content in mononuclear cells from peripheral blood (PBMNCs) of these patients and whether or not this content is modified with age. METHOD: PBMNCs of 48 patients with DS and 48 healthy controls were studied. RNA content of isolated PBMNCs was evaluated by flow cytometric measurements. RESULTS: Average RNA content of younger DS patients' cells was significantly higher than that of healthy controls (P=0.003). Furthermore, the RNA content decreased significantly with increasing age of DS patients (r=-0.377, P=0.008) in the range of 0-26 year old, whereas no significant relationship was found between age and PBMNCs' RNA content of healthy controls in the same range of ages. CONCLUSION: RNA content of PBMNCs from DS patients decreases rapidly with age. This is the first work on the age-dependent decrease of the RNA content in PBMNCs of DS patients. PMID- 16353215 TI - Report on the second Latin American consensus conference for flow cytometric immunophenotyping of hematological malignancies. AB - On May 3 and 4, 2005, the Second Latin American Consensus Conference for the Immunophenotyping of Hematological Malignancies took place in Queretaro, Mexico, with representatives from 10 countries of the region and two external consultants. This document summarizes the major conclusions for which consensus were achieved. Major differences regarding the recommendations from the first conference, which took place 9 years ago, concern the medical indications and the antibody panels for immunophenotyping. The aim of disseminating these guidelines to the international community is based on the potential interest for other countries with similar socioeconomic conditions. PMID- 16353216 TI - Epithelioid sarcoma in children and adolescents: a report from the Italian Soft Tissue Sarcoma Committee. AB - BACKGROUND: Epithelioid sarcoma (ES) is an uncommon malignant soft tissue tumor. To the authors' knowledge, little information is available to date concerning its clinical features and management in children and adolescents, particularly with regard to the recently described proximal-type variant. The current study concerns 30 patients age<18 years who were enrolled in the Italian Soft Tissue Sarcoma Committee protocols. METHODS: Histopathologic specimens, clinical data, and treatment modalities were reviewed for the current analysis. Nineteen patients had classic ES and 11 had proximal-type ES. Surgery was the mainstay of treatment; radiotherapy was given to six patients considered to be at risk of local disease recurrence due to incomplete resection, and chemotherapy was administered to eight patients. RESULTS: With a median follow-up of 66 months, the 5-year event-free survival (EFS) and overall survival (OS) rates were 61.7% and 92.4%, respectively, but the OS rate dropped to 86.9% and 72.4%, respectively, at 10 years and 15 years. Local disease recurrence was the major cause of treatment failure. The most significant finding influencing both EFS and OS was tumor site, with a tumor location in the extremities predicting a favorable outcome. Initially unfavorable clinical findings and a worse outcome were associated with the proximal-type variant of ES. A response to chemotherapy was noted in three of the seven patients with measurable disease. CONCLUSIONS: The current study confirms some typical features of ES (i.e., the peculiar superficial distal location [i.e., the hand, fingers], indolent growth, and a tendency to recur locally). The current study data do not clearly confirm the strong tendency for the lymph node involvement described in adult ES patients. Further studies are needed to better define the clinical behavior and biology of the proximal-type variant of ES. PMID- 16353217 TI - mtDNA variation in Inuit populations of Greenland and Canada: migration history and population structure. AB - We examined 395 mtDNA control-region sequences from Greenlandic Inuit and Canadian Kitikmeot Inuit with the aim of shedding light on the migration history that underlies the present geographic patterns of genetic variation at this locus in the Arctic. In line with previous studies, we found that Inuit populations carry only sequences belonging to haplotype clusters A2 and D3. However, a comparison of Arctic populations from Siberia, Canada, and Greenland revealed considerable differences in the frequencies of these haplotypes. Moreover, large sample sizes and regional information about birthplaces of maternal grandmothers permitted the detection of notable differences in the distribution of haplotypes among subpopulations within Greenland. Our results cast doubt on the prevailing hypothesis that contemporary Inuit trace their all of their ancestry to so-called Thule groups that expanded from Alaska about 800-1,000 years ago. In particular, discrepancies in mutational divergence between the Inuit populations and their putative source mtDNA pool in Siberia/Alaska for the two predominant haplotype clusters, A2a and A2b, are more consistent with the possibility that expanding Thule groups encountered and interbred with existing Dorset populations in Canada and Greenland. PMID- 16353218 TI - Anatomy of a serial killer: differential diagnosis of tuberculosis based on rib lesions of adult individuals from the Coimbra Identified Skeletal Collection, Portugal. AB - The role of new bone formation on visceral surfaces of ribs in the diagnosis of tuberculosis (TB) in past human populations has been explored by many researchers, using both skeletal remains with known causes of death and archaeological samples. This study focuses, firstly, on adult skeletons from the Coimbra Identified Skeletal Collection in Portugal and investigates the skeletal manifestations of individuals known to have died from TB; secondly, this study focuses on the role of rib lesions in the diagnostic criteria for TB. One hundred and fifty-seven males and 106 females aged between 22-87 years were examined; causes of death were assigned as pulmonary TB, extrapulmonary TB, and pulmonary non-TB; a control group, extrapulmonary non-TB, was selected from the remaining individuals. Of individuals with rib lesions, 85.7% (69/81) had pulmonary or extrapulmonary TB as an assigned cause of death, while 17.8% (16/90) of individuals with rib lesions had a non-TB cause of death. Rib lesions were significantly more common in individuals who had died from TB, although the lesions cannot be considered pathognomonic for TB. In individuals dying from pulmonary TB, ribs in the central part of the rib cage were most affected, at their vertebral ends. The lower part of the rib cage may be a marker for peritoneal TB, and "coral-like" new bone formation on ribs may be an indicator of neoplastic disease. Further work on rib involvement in TB in clinical contexts, and the study of further documented skeletal collections, are recommended. PMID- 16353219 TI - Health of the non-elites at Tombos: Nutritional and disease stress in New Kingdom Nubia. AB - During the New Kingdom period, Egypt succeeded in occupying most of Nubia. Colonial towns were built, which served as centers of government and redistribution. This paper uses a bioarchaeological approach to address the effects of this cultural contact on non-elites. Skeletal remains from the site of Tombos (N = 100), a cemetery in Upper Nubia dating to this important time, are analyzed, in addition to 1,082 individuals from contemporaneous Egyptian and Nubian sites, in order to shed light on the social, political, and economic processes at play and to determine how the people at Tombos were affected during this transitional period. In many ways, the Tombos population appears to have been affected by similar stressors as the other populations under study. However, a few small differences in the subadult frequencies of pathological lesions, especially remodeling rates, are significant in the overall picture of health at Tombos. These analyses suggest that, although the people of Tombos may have been integrated into the Egyptian colonial network, the additional resources they may have obtained could not protect them from nutritional and disease stress. A lower childhood survival through bouts of ill health at Tombos is suggested. While status may have played a role in the differences seen in the comparative populations, it is likely that parasites and/or other infections led to childhood illness and death. PMID- 16353220 TI - Twinning and heteropaternity in chimpanzees (Pan troglodytes). AB - Unlike monozygotic (MZ) twins, dizygotic (DZ) twins develop from separate ova. The resulting twins can have different sires if the fertilizing sperm comes from different males. Routine paternity testing of a pair of same-sexed chimpanzee twins born to a female housed with two males indicated that the twins were sired by two different males. DNA typing of 22 short-tandem repeat (STR) loci demonstrated that these twins were not MZ twins but heteropaternal DZ twins. Reproductive data from 1926-2002 at five domestic chimpanzee colonies, including 52 twins and two triplets in 1,865 maternities, were used to estimate total twinning rates and the MZ and DZ components. The average chimpanzee MZ twinning rate (0.43%) equaled the average human MZ rate (0.48%). However, the chimpanzee DZ twinning rate (2.36%) was over twice the human average, and higher than all but the fertility-enhanced human populations of Nigeria. Similarly high twinning rates among African chimpanzees indicated that these estimates were not artifacts of captivity. Log-linear analyses of maternal and paternal effects on recurrent twinning indicated that females who twinned previously had recurrence risks five times greater than average, while evidence for a paternal twinning effect was weak. Chimpanzee twinning rates appear to be elevated relative to corresponding estimated human rates, making twinning and possibly heteropaternity more important features of chimpanzee reproductive biology than previously recognized. PMID- 16353221 TI - Interpersonal violence in prehistoric San Pedro de Atacama, Chile: behavioral implications of environmental stress. AB - The prehistoric population of San Pedro de Atacama lived through periods marked by prosperity and interregional interaction, as well as times of severe drought, social stress, and widespread poverty. A sample of 682 crania was analyzed for evidence of cranial trauma in order to assess changing patterns of interpersonal violence during the occupation of the oasis. It was hypothesized that the level of traumatic injuries in this population would parallel some of the changes seen in the archaeological record. Low fracture rates would be expected in periods of affluence and environmental stability, while periods characterized by environmental extremes and state collapse would yield elevated rates of aggression. This analysis found that rates of trauma escalated from 5.1% (5/99) in the earliest period, to 10.9% (10/92) in the Middle Horizon (AD 600-950). Although it may reflect problems related to increasing population density in the oasis, this increase is surprising, given that the early period witnessed the shift to permanent settlements, and the middle period was one of prosperity and plentiful resource availability. Trauma rates peaked at 35.6% (16/45) in an early Late Intermediate period (AD 950-1400) cemetery, with other Late Intermediate cemeteries demonstrating similarly high rates of traumatic injury. The elevated trauma rates during this period correlate with major droughts, the concentration of settlements on the oasis' east side, fortified structures, and material poverty, all reflected in the archaeological record. As the Late Intermediate waned and environmental conditions improved, trauma concomitantly decreased (7.0%), and remained low throughout the Inka occupation (AD 1400-1532). This indicates that while the Atacama was not peaceful, violence became commonplace only during periods of great social change and resource stress. PMID- 16353222 TI - Computerized restoration of nonhomogeneous deformation of a fossil cranium based on bilateral symmetry. AB - We developed a computerized method of correcting plastic deformation of a fossil skull, based on bilateral symmetry with respect to the midsagittal plane, and applied this method to reconstruction of a fossilized Proconsul heseloni cranium (KNM-RU-7290A). A three-dimensional (3D) model of the fossil was generated using consecutive cross-sectional images retrieved from computed tomography. 3D coordinates of anatomical landmarks that should be located on the midsagittal plane and pairs of landmarks that should be symmetrical with respect to this plane were acquired. These landmarks were then repositioned so that geometrical constraints were satisfied, while translated distances of landmarks were minimized. We adopted a thin-plate spline function to mathematically describe the 3D nonlinear volumetric transformation between acquired and repositioned landmarks. Using this function, the entire fossil shape was transformed, and the effect of reversing the deformation could be visualized. The results indicated that the proposed method was effective in eliminating nonhomogeneous deformation of the fossil skull. The antemortem appearance of the skull cannot be completely restored by this method alone, due to methodological limitations. However, the presented method has a role as an adjunct in complementing conventional restoration techniques on account of its objective nature. PMID- 16353223 TI - Male competition and paternity in wild chimpanzees of the Tai forest. AB - In social animals, competition among males for mates affects individual reproductive success. The priority-of-access model attempts to account for the influence of demographic conditions within groups upon male reproductive success, but empirical data for testing this model are scarce. Our long-term study of chimpanzees in the Tai National Park, Cote d'Ivoire, encompasses a period of steady decrease in community size and fluctuating numbers of competing males and sexually receptive females. These demographic changes, in combination with genetic assessment of paternity for 48 offspring from three communities, allowed us to quantify the effects of varying levels of competition upon male reproductive success. On average, the highest-ranking male sired 50% of all analyzed offspring during a 14-year period from 1987-2000. Competition among males strongly decreased the relative reproductive success of the alpha male, such that the alpha male's rate of success decreased from 67% with few competitors to only 38% with four or more competitors. The increasing number of synchronously receptive females in large groups also reduced the proportion of paternities by the alpha male. Thus, patterns of paternity in Tai chimpanzees fit well the predictions of the priority-of-access model. We also found that despite the inability of dominants to monopolize reproduction, they achieved a higher reproductive rate in large multimale groups, because these have more females and a higher infant survival rate. Varied levels of male competition within communities seem to explain differences in the reproductive success of alpha males observed in different chimpanzee populations, and in other primate species. PMID- 16353224 TI - Reevaluation of dominance hierarchy in bonobos (Pan paniscus). AB - While dominance relationships have been widely studied in chimpanzees, in bonobos, dominance style and linearity of hierarchy are still under debate. In fact, some authors stated that bonobo hierarchy is nonlinear/ill-defined, while others claimed that it is fairly linear. In this paper, we test the hypothesis that a shift in group composition determines changes in linearity of hierarchy. To test this hypothesis, we collected data on one of the largest captive groups in the world, in the Apenheul Primate Park (The Netherlands). We investigated the linearity of the hierarchy in two different periods, with a shifting group composition. We used the corrected Landau's index and David's scores to estimate which animals were most dominant. The major overall result of our study is that hierarchy is fairly nonlinear in this group: during the first study period (eight adults), the hierarchy was nonlinear, whereas during the second one (six adults), it failed to reach statistical linearity. We argue that the reduction of the number of adults is the principal factor affecting linearity. We also found that dominance interactions were evenly distributed across sex classes in both study periods. Furthermore, no correlation was observed between age/body weight and rank. As for the overall dominance relationship between males and females, our results suggest that there is no exclusive female dominance in the Apenheul group. The dominance style of bonobos may be loose and differentially expressed in diverse groups or in the same group, along with shifting conditions. PMID- 16353225 TI - Tooth wear and dental pathology at the advent of agriculture: new evidence from the Levant. AB - Differences in patterns of diet and subsistence through the analysis of dental pathology and tooth wear were studied in skeletal populations of Natufian hunter gatherers (10,500-8300 BC) and Neolithic populations (8300-5500 BC, noncalibrated) from the southern Levant. 1,160 Natufians and 804 Neolithic teeth were examined for rate of attrition, caries, antemortem tooth loss, calculus, periapical lesions, and periodontal processes. While the Natufian people manifest a higher rate of dental attrition and periodontal disease (36.4% vs. 19%), Neolithic people show a higher rate of calculus. Both populations manifested low and similar rates of caries (6.4% in the Natufian vs. 6.7% in the Neolithic), periapical lesions (not over 1.5%), and antemortem tooth loss (3.7% vs. 4.5%, respectively). Molar wear pattern in the Neolithic is different than in the Natufian. The current study shows that the dental picture obtained from the two populations is multifactorial in nature, and not exclusively of dietary origin, i.e., the higher rate and unique pattern of attrition seen in the Natufian could result from a greater consumption of fibrous plants, the use of pestles and mortars (which introduce large quantities of stone-dust to the food), and/or the use of teeth as a "third hand." The two major conclusions of this study are: 1) The transition from hunting and gathering to a food-producing economy in the Levant did not promote changes in dental health, as previously believed. This generally indicates that the Natufians and Neolithic people of the Levant may have differed in their ecosystem management (i.e., gathering vs. growing grains), but not in the type of food consumed. 2) Changes in food-preparation techniques and nondietary usage of the teeth explain much of the variation in tooth condition in populations before and after the agricultural revolution. PMID- 16353226 TI - Sonography of the first dorsal metatarsal artery of the foot. AB - PURPOSE: To describe the gray-scale and Doppler sonographic examination technique and appearances of the first dorsal metatarsal artery (FDMA) of the foot and discuss its clinical significance. METHODS: The dorsal arteries of foot-including the FDMA and the first plantar metatarsal artery-of 374 feet were studied using gray-scale and color Doppler sonography. Due to the difficulty of detecting the FDMA, a majority of them were identified by studying the web area just distal to the intermetatarsal space. The interosseous muscle was identified to determine the classification of the anatomic type of the FDMA. The sonographic results of 152 feet were compared with surgical findings. RESULTS: The average caliber of the FDMA was 1.0 +/- 0.5 mm. The FDMA was classified into 3 groups of variants based on its relationship with the first dorsal interosseous muscle. The groups were identified as the superficial artery (54.5%), intramuscular artery (39.6%), and submuscular artery (9%) variants. The rates of visualization of the distal and proximal portions of the FDMA were 86.6% and 47%, respectively. In the first intermetatarsal space, there were anastomoses between the FDMA and the first plantar metatarsal arteries in 87.6% of the feet. CONCLUSIONS: The FDMA varies greatly; however, sonography can clearly show its morphologic characteristics, such as caliber and location. Sonography also provides valuable preoperative information in microsurgery of the foot. PMID- 16353227 TI - Power Doppler assessment of overall disease activity in patients with rheumatoid arthritis. AB - PURPOSE: To examine synovial vascularity and flow patterns in hand and wrist joints--metacarpophalangeal (MCP) joints and ulnar stiloid (USTL) regions--of patients with rheumatoid arthritis (RA) using power Doppler sonography (PDUS) and spectral Doppler analysis and to assess the accuracy of PDUS in detecting overall disease activity in RA patients. METHODS: Two hundred forty MCP joints and 48 USTL regions in 24 RA patients were examined. Patients were categorized into 2 groups--active and inactive--according to the American College of Rheumatology remission criteria. Resistance indexes (RIs) were measured. RESULTS: Flow signals were detected in 50 MCP joints (in 13 patients) and 24 USTL regions (in 16 patients) and spectral analysis was performed in 46 MCP joints (12 patients) and 23 USTL regions (16 patients). The sensitivity and specificity of PDUS in detecting disease activity in RA were 92% and 40%, respectively. There was a negative correlation between flow signal number and RI, with higher scores of flow signals corresponding to lower RIs. CONCLUSION: PDUS appears to be a reliable method for assessing inflammatory activity in rheumatoid synovium. PMID- 16353228 TI - Plantar fasciitis treated with local steroid injection: comparison between sonographic and palpation guidance. AB - PURPOSE: To compare the effectiveness of sonographically guided and palpation guided steroid injection for the treatment of proximal plantar fasciitis. PATIENTS AND METHODS: Twenty-five consecutive patients with unilateral proximal plantar fasciitis were recruited and randomly divided into a sonographically guided group (n = 12) and palpation-guided group (n = 13). Proximal plantar fascia was assessed with a 5- to 12-MHz linear-array transducer. Pain intensity was quantified using a "tenderness threshold" (TT) and a visual analog scale (VAS). Injection of 7 mg (1 ml) of betamethasone and 0.5 ml of 1% lidocaine into the inflamed proximal plantar fascia was performed under the guidance of sonography or palpation. Patients were evaluated clinically and sonographically before injection and at 2 weeks, 2 months, and 1 year after injection. VAS- and TT-measured pain intensity, thickness, and echogenicity of the proximal plantar fascia, as well as the recurrence of heel pain, were assessed. RESULTS: Both VAS- and TT-measured levels of pain improved significantly after steroid injection in both groups (p < 0.001). Also, the thickness decreased significantly after injection (p < 0.01 in the palpation-guided group; p < 0.001 in the sonographically guided group). The number of patients with hypoechogenicity at the proximal plantar fascia decreased after steroid injection in both groups (p < 0.01 for both groups). The recurrence rate of plantar fasciitis in patients of the palpation-guided group (6/13) was significantly higher than that of the sonographically guided group (1/12) (p < 0.05). CONCLUSIONS: Steroid injection can be an effective way to treat plantar fasciitis, and injection under sonographic guidance is associated with lower recurrence of heel pain. PMID- 16353229 TI - The influence of athletic activity on the plantar fascia in healthy young adults. AB - PURPOSE: Complaints deriving from the plantar fascia are relatively common in athletes. This study aimed to investigate the changes of thickness of plantar fascia via sonography in healthy young adults with different levels of activity. PATIENTS AND METHODS: One hundred ten adults with normal body mass index were separated into three groups according to activity level: sedentary (group 1, n = 50), athletic activity less than 7 hours per week (group 2, n = 30), and athletic activity 7 or more hours per week (group 3, n = 30). The thicknesses of the plantar fascia at origin and at a point 5 mm distal to origin were measured via sonography. RESULTS: The mean values of the thickness of the proximal plantar fascia (PFp) and the distal plantar fascia (PFd) in group 1 were similar to those of groups 2 and 3 (p > 0.05). The mean values of PFp and PFd were significantly higher in men than in women (p < 0.05). The mean values of PFp and PFd were similar in left and right feet (p > 0.05). There were moderate positive correlations between PFp and weight, height, and body mass index but no correlation between PFp and amount of athletic activity. CONCLUSIONS: The thickness of the plantar fascia at origin did not change with athletic activity at the amateur level. PMID- 16353230 TI - Incidence and clinical significance of spontaneous intrahepatic portosystemic venous shunts detected by sonography in adults without potential cause. AB - PURPOSE: B-mode and color Doppler sonography were used to assess the incidence and clinical significance of spontaneous intrahepatic portosystemic venous shunt in adults without detectable cause. METHODS: A total of 25,579 adults without potential cause of intrahepatic portosystemic venous shunt were screened using B mode sonography. Suspicion of shunt was confirmed by color Doppler sonography. Patients with shunt were followed up with sonography at 6- to 12-month intervals and received dietary education to prevent the ingestion of extra-large amounts of protein. During each sonography session, the patients were asked to report any experience of consciousness disturbance. RESULTS: Six patients (0.0235%) showed evidence of shunt on color Doppler sonography. Direct communication between a right portal branch and the inferior vena cava was found in 3 patients; a small aneurysm connecting a peripheral portal branch to the hepatic vein was found in the remaining 3 patients. Five patients were followed up (range, 12-60 months) and showed no change in the size of the shunt. None of the patients with shunt reported an experience of consciousness disturbance. CONCLUSIONS: The incidence of spontaneous intrahepatic portosystemic venous shunt is extremely low in asymptomatic adult patients. Follow-up and dietary control seen to be suitable for their management. PMID- 16353231 TI - Unusually asymmetric venous engorgement of the breast after long-term hemodialysis. AB - Venous engorgement of the breast demonstrated on mammography is usually considered a minor sign of malignancy. We report the sonographic and mammographic findings in an unusual case of asymmetric enlargement of the mammary veins in a middle-aged patient as a complication of long-term hemodialysis. PMID- 16353232 TI - Androgen insensitivity syndrome: in utero diagnosis by four-dimensional sonography and amniotic fluid karyotype. AB - We describe a fetus with androgen insensitivity syndrome diagnosed at 16 weeks' gestation with two-dimensional (2D) and four-dimensional (4D) sonography and karyotype analysis in a 37-year-old pregnant woman. Two-dimensional (2D) sonography revealed a female phenotype. Karyotype analysis revealed an unremarkable 46,XY chromosomal complement. Repeat 4D sonography confirmed a female phenotype with edematous labia majora, consistent with androgen insensitivity syndrome. PMID- 16353233 TI - Sequential bilateral torsion of normal ovaries in a prepubertal child. AB - We report a case of bilateral ovarian torsion in a prepubertal girl with normal ovaries that occurred at an interval of 1 year. The diagnosis was made on gray scale and color Doppler sonography, and ovarian salvage was performed. The role of preventive contralateral ovarian pexy in cases of ovarian torsion is emphasized. PMID- 16353234 TI - Peptic ulcer perforation: sonographic imaging of active fluid leakage. AB - Sonography is not the method of choice for the evaluation of suspected peptic ulcer perforation (PUP). However, indirect sonographic signs and direct visualization of PUP have been reported by several authors in recent years. We report a case of an elderly woman who presented with severe abdominal pain and positive rebound sign, in whom abdominal sonography demonstrated indirect signs of PUP, the site of perforation, and active air fluid leakage through the perforated anterior prepyloric antral wall. PMID- 16353235 TI - Triplication of 8p22-8p23 in a patient with features similar to Kabuki syndrome. AB - Kabuki syndrome (KS) comprises multiple congenital anomalies and distinctive facial appearance. Although a number of chromosome abnormalities have been described in patients with KS-like phenotypes, no consensus has been reached regarding the genetic basis underlying the classic Kabuki phenotype. A recent study reported on 8p22-8p23.1 duplication in patients diagnosed with KS; however, a number of other studies have not found this duplication in patients with classic KS. We report on a girl with triplication of 8p22-8p23 who has mental retardation and some features suggestive of KS, including growth retardation, left-sided obstructive heart lesion, long-appearing palpebral fissures, hypertelorism, sparse lateral eyebrows, prominent ears, and persistent fetal fingertip pads. She does not have the typical facial gestalt of KS, nor does she have other more specific findings of KS. We propose that abnormal copy number of genes in the 8p22-8p23 region results in a syndrome of multiple congenital anomalies with many features that overlap with classic KS. However, data from this patient and others with similar duplications in the literature suggest that duplication or triplication of 8p22-8p23 represents a recognizable pattern of malformation distinct from classic KS. The exact genetic abnormality underlying KS currently remains unknown. PMID- 16353236 TI - Fetal alcohol spectrum disorders in Finland: clinical delineation of 77 older children and adolescents. AB - The adverse effects of alcohol on the developing human comprise a spectrum of structural anomalies and behavioral and neurocognitive disabilities, most accurately termed fetal alcohol spectrum disorders (FASD). We previously have proposed revisions to the 1996 Institute of Medicine Diagnostic Criteria for diagnoses in the FASD continuum [fetal alcohol syndrome (FAS), partial fetal alcohol syndrome (PFAS), alcohol related birth defects (ARBD), and alcohol related neurodevelopmental disorder (ARND)], allowing for more reproducible and accurate FASD diagnosis in a clinical setting [Hoyme et al., 2005]. The NIAAA recently has coordinated and funded an international consortium of projects aimed at more complete characterization of the teratogenic spectrum of alcohol. One of the projects sites is in Finland. The aims of this project are: (1) to completely clinically characterize the structural and learning/behavioral phenotypes of a large cohort of older children and adolescents with moderate to severe disability within the FASD continuum; (2) to correlate FASD dysmorphology and behavioral phenotypes with CNS structure and function (i.e., MRS, MRI correlations); (3) to compare the phenotype of a genetically homogeneous population of Finnish children with FASD to that observed in other populations. We have recently completed dysmorphology examination and parent/guardian interviews of the 77 children in the Finnish cohort. The purpose of this report is to present historical and morphometric data on these patients, thereby more completely delineating the clinical spectrum of FASD in older children and adolescents, contrasting the phenotype with that described in other populations and examining whether a weighted dysmorphology score could be used as a clinical and research adjunct when fetal alcohol exposure is being suspected. All children were previously diagnosed with FASD by an experienced pediatric specialist in Finland, and all were exposed to significant maternal alcohol abuse prenatally. The sex ratio of the cohort was 0.38 (male: female) and ages ranged from 8 to 20 years, with a mean of 13 years. After application of the Revised IOM Diagnostic Criteria, 53% of the subjects were diagnosed as having FAS, 30% PFAS, 12% ARND, and 5% other diagnoses. Of note, although a family history of mental retardation or birth defects was rare, 43% of the children had one or more sibling who also carried a diagnosis of FAS. Eighty-nine percent of the mothers smoked cigarettes during gestation; other teratogenic exposures were rare. Almost none had undergone genetics evaluation in the past. Almost all of the subjects had resided in multiple foster placements since early childhood and had been followed regularly by pediatric specialists. Although 11% were born prematurely, 70% demonstrated prenatal growth deficiency, and 45% were microcephalic. Other than growth deficits and the cardinal facial features, the most common major and minor anomalies noted were: camptodactyly (55%), "hockey stick" or other altered palmar creases (51%), refractive errors (40%), strabismus (38%), dental crowding (43%), nail hypoplasia (38%), GU anomalies (22%), and congenital heart defects (18%), "Railroad track" ears were not observed in this population. PMID- 16353237 TI - Theaflavin-3, 3'-digallate induces epidermal growth factor receptor downregulation. AB - Black tea is one of the most popular beverages worldwide and especially in Western nations. Theaflavins, a mixture of theaflavin (TF-1), theaflavin-3 gallate (TF-2a), theaflavin-3'-gallate (TF-2b), and theaflavin-3,3'-digallate (TF 3) are the major components of black tea. Among these black tea components, theaflavin is generally considered to be the more effective component for the inhibition of carcinogenesis. Recently, TF-3 has been shown to have an antiproliferative effect on tumor cells, but the mechanism is not clear. In this study, we showed that TF-3-induced internalization and downregulation of the epidermal growth factor receptor (EGFR). These results suggested that TF-3 induces EGFR endocytosis and degradation. We further showed that TF-3 stimulated EGFR ubiquitination and tyrosine kinase activation. Interestingly, TF-3-induced EGFR downregulation is inhibited by the proteasome inhibitor, MG132, but not by the EGFR-specific receptor tyrosine kinase inhibitor, AG1478. Furthermore, pretreatment with TF-3 inhibited EGF-induced EGFR autophosphorylation, ERKs phosphorylation and AP-1 activation in JB6 Cl41 cells. In addition, TF-3 inhibited EGF-induced anchorage-independent cell transformation. Overall, our results indicate that TF-3 might exert chemopreventive effects through the downregulation of the EGFR. PMID- 16353238 TI - 1-methyl-4-phenylpyridinium neurotoxicity is attenuated by adenoviral gene transfer of human Cu/Zn superoxide dismutase. AB - Oxidative stress has been suggested to be an important mediator of dopaminergic cell death in Parkinson's disease (PD). We investigated the neuroprotective potential of Cu/Zn superoxide dismutase (SOD1) overexpression in the rat substantia nigra (SN) following adenovirus-mediated gene transfer. Human dopaminergic SK-N-SH cells were transduced with adenoviral vectors expressing either human SOD1 (Ad-SOD1) or beta-galactosidase (Ad-betagal) before exposure to 1 mM of the 1-methyl-4-phenylpyridinium ion (MPP+). A strong neuroprotective effect of SOD1 gene transfer was observed in the SK-N-SH cells exposed to MPP+ compared with controls. Adult rats were then given unilateral injections of either Ad-SOD1 or Ad-betagal into the striatum, and MPP+ was administered 8 days later at the same location. Strong transgene expression was detected in the SN dopaminergic neurons, a consequence of retrograde axonal transport of the adenoviral particles. The amphetamine-induced rotational behavior of the rats was markedly lower in Ad-SOD1-injected rats than in control animals. Also, behavioral recovery significantly correlated with the number of tyrosine hydrolase expressing neurons in the SN of the treated rats. These results are consistent with oxidative stress contributing to the MPP+ -induced neurodegenerative process. They also indicate that SOD1 gene transfer into the nigrostriatal system may be a potential neuroprotective strategy for treating PD. PMID- 16353239 TI - What are the supportive structures of the female urethra? AB - AIMS: Female stress urinary incontinence is thought to result from impairment of the connective tissue "ligaments" of the urethra. Surgical repair of female incontinence mainly involves fixation of the urethra to the pubic bone or other surrounding structures. In the present anatomical-radiological study, the anatomy of the connective tissue structures around the female urethra was investigated to determine the anatomical structures that support the urethra and the rhabdosphincter. MATERIALS AND METHODS: The topography of the anterior compartment of the female pelvis was studied in serial sections and one anatomical preparation of 30 female fetuses and of six adult females. The pelves of 29 female fetuses were processed according to plastination histology technique. The pelves of the six adult specimens were processed according to sheet plastination technique. In addition, the anatomical findings were compared with MR images of 41 adult female volunteers. RESULTS: The ventro-lateral aspect of the urethra remains free of fixating ligaments throughout its pelvic course. Ventro-laterally the urethra is enclosed by the ventral parts of the levator ani, its fasciae and a ventral urethral connective tissue bridge connecting both sides. Dorsally, the urethra is intimately connected to the wall of the vagina. CONCLUSIONS: The female urethra has no direct ligamentous fixation to the pubic bone. Urethral continence after pregnancy and childbirth may be explained by a widening of the hiatus of the levator ani or the anterior vaginal wall, resulting in overstretching of the ventral urethral connective tissue bridge or the disruption of the fixation between urethra and vagina. PMID- 16353240 TI - Introduction to Judith Hall Festschrift. PMID- 16353241 TI - Bosma arhinia microphthalmia syndrome. AB - Bosma et al. [1981] delineated a syndrome affecting two unrelated males with severe hypoplasia of the nose and eyes, palatal abnormalities, deficient taste and smell, inguinal hernias, hypogonadotropic hypogonadism with cryptorchidism, and normal intelligence. Other patients with this syndrome have been reported as cases of arhinia with associated defects. During embryonic development, the nasal placodes form 28 days after conception shortly after the optic vesicles makes contact with the overlying surface ectoderm at 26-27 days, and both layers invaginate to form the eyes between 34 and 44 days. Mice with homozygous mutations of Pax6, manifest underdevelopment of ocular and nasal structures, and a network of developmentally regulated genes function downstream of Pax6 to form nasal, ocular, and pituitary structures. These genes represent candidate genes for this disorder, and familial recurrence of Bosma syndrome has been reported to occur. This report describes two sporadic unrelated cases of this rare syndrome and briefly reviews the findings in previously reported cases. PMID- 16353242 TI - Festschrift reflection. PMID- 16353243 TI - Mitochondrial mutation in a child with distal arthrogryposis. AB - A 15-year-old girl presented with stroke and was found to have a mitochondrial mutation MELAS T3271C. As an infant, she had required casting and surgery for clubbed feet as well as physical therapy for camptodactyly and adducted thumbs. Despite some residual deformities, psychomotor development was normal, but she was extremely small for her age with height, weight, and head circumference below the third centile. A younger brother had similar distal arthrogryposis, but normal height and weight and no history of stroke though he had been diagnosed with attention deficit. The parents and another brother were clinically normal. While the observation of MELAS in a child with distal arthrogryposis could be co incidental, it raises concern about the possible role of mitochondrial myopathy or neuropathy in causation of distal arthrogryposis. PMID- 16353244 TI - Familial complex 3q;10q rearrangement unraveled by subtelomeric FISH analysis. AB - In recent years, subtelomeric rearrangements have been identified as a major cause of multiple congenital anomalies/mental retardation syndromes. Currently, more than 2,500 individuals with mental retardation have been tested and reported in whom subtelomeric rearrangements were detected ranging from 2% to 29%. Therefore, subtelomeric FISH analysis is indicated as a second tier test after high-resolution G-banding analysis in patients with otherwise unexplained developmental delay/mental retardation and/or multiple congenital anomalies. We describe a patient and her three maternal female cousins, all showing an undiagnosed MCA/MR syndrome, associated with the same complex subtelomeric rearrangement. Subtelomeric FISH testing performed between 3(1/2) and 18 years after the initial karyotype showed, in all four patients, distal trisomy 3q and distal monosomy 10q as follows: 46,XX,ish der(10)t(3;10)(q29;q26.3)mat(D10S2488+,D10S2490-, D3S1272+,D10Z1+). Parental subtelomeric FISH analysis showed that the proposita's mother and three of four brothers and one of two sisters had a cryptic balanced 3:10 telomere translocation. The three brothers with the balanced translocation were father to one each of the three proband's cousins. All four affected girls showed a similar phenotype with pre/postnatal growth retardation, microcephaly, severe developmental delay/mental retardation, poor/absent speech, and a distinct pattern of malformation. On examination there were coarsening of facial features with low fronto-temporal hairline; thick eyebrows; bilateral epicanthal folds; hypertelorism; prominent nose with squared nasal root and narrow alar base; low set posteriorly rotated large ears with a prominent anthelix; high arched palate; prominent chin; hands/feet brachydactyly; bilateral squint; hypotonia; and muscle hypotrophy. A slow overall improvement was seen in all patients over time. To our knowledge, this complex subtelomeric rearrangement in our patients has never been reported so far. Monosomy 10q has recently been described either isolated or as part of a complex rearrangement involving telomeres other than the 3q. Trisomy 3q29 has not yet been reported, but our patients resembled cases with 3q26 trisomy suggesting that the critical region of duplication for this phenotype is in 3q29. PMID- 16353245 TI - Meckel on developmental pathology. AB - Before Schleiden and Schwann, Darwin and Mendel there passed briefly a towering giant, Johann Friedrich Meckel the Younger (1781-1833), now glimpsed only fleetingly and obscurely through the mist of time and former controversies, who can nowadays easily and clearly be identified as the father of a "pre-modern" developmental biology. At his beginning this prodigiously gifted physician scholar had, as one would say nowadays, an unfair advantage, his cradle having been rocked, as it were, by the preparators in his father's and grandfather's huge collection of normal and abnormal anatomical "specimens" in the home in which he was born and raised including his father's own skeleton (with two anatomical anomalies!). Initially reluctant to follow in the steps of his illustrious anatomist/physician grandfather and father, he nevertheless early demonstrated extraordinary gifts in anatomy and zootomy. Napoleon's conquest of his homeland notwithstanding, Meckel spent at least 2 extremely fruitful years in Paris, under the tutelage of Cuvier, but also in close contact with Geoffroy St. Hilaire (Etienne), Lamarck, and von Humboldt. He not only translated Cuvier's Lecons d'anatomie comparee into German but also greatly enriched this pivotal treatise with observations of embryonic and malformed fetuses and animals only of passing interest to his mentor. In his numerous publications, Meckel was the first to relate abnormal to normal development, define anomalies of incomplete differentiation (vestigia), but, most importantly, to relate those malformations known in humans to those that are normal adult developmental states in "lower" animals (atavisms). Thus, Meckel's three-fold parallelism of the scala naturae, normal ontogeny, and the malformations in humans and animals makes him a recapitulationist par excellence, however, without ever venturing into a fully articulated and explicit theory of descent. Today Meckel is remembered solely as the discoverer of the syndrome and cartilage named after him, and as having interpreted, correctly, the developmental nature of the "Meckel" diverticulum. It is virtually unknown that Meckel also first enuntiated the concept and distinction between primary and secondary malformations/anomalies, introduced the notion of heredity into the causal analysis of congenital anomalies, was the father of syndromology (the Meckel syndrome), had a clear understanding of pleiotropy and heterogeneity, and can unequivocally be regarded as the father of developmental pathology. In hindsight, and inspite of much professional success, Meckel emerges as a tragic figure in the history of biology, his life cut short at 52 without an ability to incorporate cell theory and the embryological insights of his younger contemporaries into his intellectual edifice which might have made it possible for him to finally and clearly see "analogy" (now homology), of which he was the greatest expert in his era, as incontrovertible evidence for descent. In that case, Darwin and Haeckel might have even had the courtesy of a tip-of-the-hat in Meckel's direction. PMID- 16353246 TI - Echocardiographic findings in classical and hypermobile Ehlers-Danlos syndromes. AB - Structural cardiovascular alterations in the classical and hypermobile forms of Ehlers-Danlos syndrome(EDS) warrant investigation. We have examined a cohort of 38 patients with hypermobile and classical EDSs using two-dimensional echocardiography. The cohort includes 7 males and 31 females, with an age range from 12-60 years. Altered echocardiographic parameters were seen in the initial cross-sectional data analysis in 24/38 patients. Five of the 38 participants had mildly dilated aortic root (AR) or sinuses of Valsalva (SV), and an additional 7 patients had an abnormal pouching of the SV, although the absolute dimensions did not exceed the normal range. Ten patients had mild mitral, tricuspid, or aortic regurgitation, and only one patient had mitral valve prolapse (MVP). Three patients had low normal systolic function; three had evidence of mildly elevated pulmonary pressures, and two patients had mild concentric left ventricular hypertrophy (LVH). Five patients had evidence of impaired left ventricular relaxation (LVR) based on mitral valve E to A velocity ratio. Interestingly, 26/38 subjects demonstrated a prominent right coronary artery (RCA) easily visualized by trans-thoracic echocardiography, and 10/38 had an elongated cardiac silhouette on the 4-chamber apical views. The "pouching" shape of the SV was more common in hypermobile type than in the classical type of EDS. The study is ongoing and will accrue longitudinal data on 100 subjects with classical and hypermobile EDSs at 2-year intervals. PMID- 16353247 TI - Serum hepatocyte growth factor activator inhibitor type I (HAI-I) and type 2 (HAI 2) in prostate cancer. AB - BACKGROUND: Hepatocyte growth factor activator inhibitor type 1 (HAI-1) and type 2 (HAI-2) are Kunitz-type serine protease inhibitors for hepatocyte growth factor activator (HGFA). We attempted to clarify whether serum levels of HAI-1 and HAI-2 could be a useful marker in patients with prostate cancer. METHODS: Serum levels of HAI-1 and HAI-2 were measured by enzyme-linked immunosorbent assay in 27 patients with benign prostatic hyperplasia (BPH) and 118 patients with prostate cancer. RESULTS: The mean serum levels of HAI-1 in patients with prostate cancer were significantly higher than those in patients with BPH. Furthermore, the serum HAI-1 levels in patients with distant metastasis and hormone resistant prostate cancer were significantly elevated compared with those in patients with organ confined diseases. There were no significant differences in serum HAI-2 levels among prostate cancer subgroups according to clinical stage. Significantly elevated levels of HAI-1 were detected in 38 patients with prostate cancer before any treatment. CONCLUSIONS: HAI-1 may be a potential tumor marker for prostate cancer. Further studies in large groups of patients are needed to define the clinical value of HAI-1. PMID- 16353248 TI - Expression of prothymosin alpha is correlated with development and progression in human prostate cancers. AB - BACKGROUND: Our previous study clearly demonstrated that decreased expression of prothymosin alpha (PTMA) was associated with inhibition of rat prostate carcinogenesis by isoflavones. The purpose of the present investigation was to provide a better understanding of the role of PTMA in human prostate cancers. METHODS AND RESULTS: PTMA expression in 68 prostate cancer cases and in prostate cancer cell lines was examined by immunohistochemistry and immunoblotting, and its levels were increased with progression from normal epithelium, through prostatic intraepithelial neoplasia (PIN) to carcinomas, correlating with the Gleason's pattern. All cell lines studied (LNCaP, 22Rv1, DU145, and PC3) showed high PTMA expression compared with prostate epithelial cells (PrEC). Knockdown of PTMA expression in PC3 cells by RNAi resulted in the inhibition of both cell growth and invasion in vitro. CONCLUSIONS: The present study clearly demonstrated that PTMA expression is intimately involved in the differentiation and progression of human prostate cancers, and could be a target for therapy and diagnostic purposes. PMID- 16353249 TI - Synthetic dimer of indole-3-carbinol: second generation diet derived anti-cancer agent in hormone sensitive prostate cancer. AB - BACKGROUND: Cruciferous vegetables have been found to have anti-prostate cancer effects. The active compounds mediating these effects include indoles such as indole-3-carbinol (I3C) and isothiocyanates. I3C is unstable having tissue tropic effects and clinical utility has been partly addressed by the synthesis of a more stable dimer diindolylmethane (DIM). METHODS: Anti-proliferative activity was measured by XTT assay and cytosolic proteins quantitated by Western blot analysis. RESULTS: DIM (IC(50) 50 microM) is a better anti-proliferative agent than I3C (IC(50) 150 microM) in androgen dependent LNCaP cells, inhibits DNA synthesis, and growth of R1881 stimulated LNCaP cells. Androgen receptor (AR), cyclin D1, and cdk4, induced by R1881, are downregulated by DIM. DIM downregulates phosphorylated Akt and phosphatidyl inositol 3-kinase and downstream inhibition of cyclin D1 and cdk4. CONCLUSION: These studies provide evidence that DIM is a second-generation chemopreventive agent with a viable cellular target and has clinical potential as an anti-prostate cancer chemopreventive. PMID- 16353250 TI - Gene therapy for prostate cancer: current strategies and new cell-based approaches. AB - Prostate cancer is the most commonly diagnosed cancer in adult males in the Western world. It accounts for one in ten cancer cases and is the second leading cause of cancer death in men, after lung cancer. A number of curative treatments are available for patients with localized prostate cancer such as radical prostatectomy, radiotherapy, or brachytherapy. However, a proportion of these men will develop progressive disease, and some will present de novo with advanced and metastatic prostate cancer, which is amenable to palliation only with androgen withdrawal therapy. Most of these patients will eventually develop hormone refractory disease which is incurable, and for whom gene therapy, if feasible may develop as an alternative treatment option. In this review we discuss the gene therapy vectors and strategies that are currently in use, new cell-based approaches, discuss their advantages and disadvantages, and review the potential or proven pre-clinical and clinical efficacy in prostate cancer models/patients. PMID- 16353251 TI - Trisomy 16p: a longitudinal profile and photo essay. AB - As more cases of complete or partial trisomy 16p are described, a clinical picture of these patients is emerging. A specific phenotype appears to be most consistent if the band 16p13.1-16p13.3 is present in triplicate. The hallmarks of this syndrome are microcephaly, a specific facial appearance with round facies, micrognathia, and small protruding auricles, and psychomotor as well as growth retardation. We report on a patient with partial trisomy 16p due to a maternally inherited balanced translocation between chromosomes 2q and 16p and describe the change in phenotype over 21 years, as well as the level of development achieved. PMID- 16353252 TI - Illustration of genetic syndromes in the nursery. AB - Reading to children and storytelling has documented developmental benefits. Traditional Nursery Rhymes (Mother Goose tales in North America) encapsulate 'snapshots' of the people described and chronicle their customs, superstitions, and amusements. Art has long been employed to document the impact of human imperfections and diseases. We investigated whether illustrations accompanying nursery rhymes, suggest that any characters illustrated may have had or been based on recognized morphological abnormalities, and if this literature documents a role for grandmothers as storytellers. Archival materials were reviewed at the Victoria and Albert museum and Mary Evans picture library, and via the web. As early as 1695, Perrault included a frontispiece of a mature woman as storyteller in his book of fairytales. Similar scenes by various artists (Boilly, Cruikshank, Guy, Highmore, Maclise, Richter, and Smith) are found consistently from 1744 to 1908. Many illustrators (Aldin, Caldecott, Cruikshank, Dore, Dulac, Gale, Greenaway, Rackham, Tarrant, and Wood) portray infants, children, and adults who are dwarfed, giant, or whimsically grotesque. Many images certainly suggest genetic syndromes, and in some characters consistency of specific features is evident between artists. Our research confirms the wealth of children's nursery rhyme illustrations suggesting pathology; that an authoritative compilation of the morphologies depicted is lacking; and that historically, grandmothers have a central role as storytellers. PMID- 16353253 TI - Recent milestones in achondroplasia research. PMID- 16353254 TI - Glycine encephalopathy (nonketotic hyperglycinemia): comments and speculations. PMID- 16353255 TI - Gene-chip analysis of the effect of tripeptide tyroservatide (YSV) on gene expression in human hepatocarcinoma BEL-7402 tumors transplanted to nude mice. AB - Tyroservatide (YSV) is a bioactive tripeptide of tyrosyl-seryl-valine. In this study, we studied the effects of YSV on human hepatocarcinoma BEL-7402 tumors transplanted in BALB/c (nu/nu) nude mice, and gene expression in the tumor cells with gene-chip analysis. Results show that YSV significantly inhibits the growth of transplanted human hepatocarcinoma BEL-7402 in nude mice (n = 12) compared with the control group (P < 0.05); with an inhibition rate of 55% at 320 microg/kg/d. Seven hundred eighty-one genes were different between the YSV group and the control group. Fifty-two genes changed in expression level by onefold or more including 37 downregulated genes and 15 upregulated genes. Probably, YSV exhibits a significant antitumor activity by inhibiting the expression of tumor cells histone genes, then damaging tumor cell chromosome and killing tumor cells. PMID- 16353256 TI - Cystic adenomatoid malformation of the lung: review of genetics, prenatal diagnosis, and in utero treatment. AB - Prenatal identification of lung abnormalities has increased with prenatal surveillance. Treatment usually requires serial ultrasound observation but in rare situations in utero therapy may be required for fetal survival. We review the genetics, prenatal evaluation, and treatment of lung abnormalities with congenital cystic adenomatoid malformation (CCAM). Other lung lesions, bronchopulmonary sequestration (BPS), hybrid lesions involving both malformations, congenital lobar emphysema (CLE), are briefly included as differential diagnosis options. Outcome of fetuses identified to have CCAM lung abnormalities resulting in fetal hydrops and having in utero therapy (thoracoamniotic shunting, fetal thoracotomy, EXIT delivery) are discussed. In the appropriate situation, this maternal fetal surgery approach for CCAM is life saving for the affected fetus with acceptable maternal morbidity risks in the present and future pregnancies. PMID- 16353257 TI - Complexity in genetic diseases: how patients inform the science by ignoring the dogma. PMID- 16353258 TI - Clinical variation of Aarskog syndrome in a large family with 2189delA in the FGD1 gene. AB - The clinical diagnosis of ASS (Aarskog-Scott syndrome or Faciogenital Dysplasia) was made in seven individuals belonging to a large Arabic family, which was supported by molecular studies revealing a 2189delA mutation in exon 15 of the FDG1 gene. The affected individuals in this family demonstrated clinical variability particularly in their cognitive skills, raising the question whether other genetic factors might be involved in the phenotypic evolution of ASS. PMID- 16353259 TI - A tribute to our teacher, Dr. Judith Hall: a child with the trait of the Earl of Shrewsbury. AB - "Organized human endeavor can be lifted an order of magnitude through teaching if it is inspiring" (Editor, Am J Dis Child, 1972). The benevolent influence of Dr. Judy Hall's inspiring clinical teaching in the field of genetic syndromes and birth defects is illustrated through the eventual surgical remediation of conductive hearing loss for a 4-year-old girl with unusual knuckles. The fascinating history of this child's syndrome has been further explored in the descendents of the first Earl of Shrewsbury. The legends of his story and his role in the Hundred Years War were immortalized by William Shakespeare in his play Henry VI Part I, but neither Shakespeare nor historians documented that the Earl actually had abnormal finger joints. Heterozygous mutations in the human noggin gene (NOG) cause a spectrum of joint fusions, including this child's traits. On behalf of practitioners of medicine, pediatrics, clinical genetics, and dysmorphology, as well as research scientists in the many domains of genetics, thank you, Judy, for your inspiration, enthusiasm, and teaching. PMID- 16353263 TI - Functional liquid-crystalline assemblies: self-organized soft materials. AB - In the 21st century, soft materials will become more important as functional materials because of their dynamic nature. Although soft materials are not as highly durable as hard materials, such as metals, ceramics, and engineering plastics, they can respond well to stimuli and the environment. The introduction of order into soft materials induces new dynamic functions. Liquid crystals are ordered soft materials consisting of self-organized molecules and can potentially be used as new functional materials for electron, ion, or molecular transporting, sensory, catalytic, optical, and bio-active materials. For this functionalization, unconventional materials design is required. Herein, we describe new approaches to the functionalization of liquid crystals and show how the design of liquid crystals formed by supramolecular assembly and nano segregation leads to the formation of a variety of new self-organized functional materials. PMID- 16353265 TI - Nitric oxide release during evoked neuronal activity in cerebellum slices: detection with platinized carbon-fiber microelectrodes. AB - Nitric oxide is an important biological messenger that particularly induces the relaxation of smooth muscle cells surrounding vessels, and, hence, controls the flow of blood. This mechanism is essential for brain function, and its fine control, termed functional hyperemia, is supposed to be realized by certain neurons that may release bursts of NO*. The aim of the present study is to examine the advantages of platinized carbon-fiber microelectrodes (5-7 microm tip diameter) for the direct and in situ electrochemical detection of NO* released by neurons into ex vivo cerebellum slices. After establishing the different analytical properties of the platinized carbon-fiber microelectrodes in vitro on NO* solutions at 50 nM to 1 mM concentration, they were characterized using DEA NONOate solutions that chemically decompose into NO*, and therefore mimic the measurement of transient variations of NO* concentration in biological samples. This validated the present approach, so that direct, in situ ex vivo measurements of nitric oxide released by neurons in a rat cerebellar slice are presented and discussed. PMID- 16353266 TI - A comparison of the fluorescence dynamics of single molecules of a green fluorescent protein: one- versus two-photon excitation. AB - We report on the dynamics of fluorescence from individual molecules of a mutant of the wild-type green fluorescent protein (GFP) from Aequorea victoria, super folder GFP (SFGFP). SFGFP is a novel and robust variant designed for in vivo high throughput screening of protein expression levels. It shows increased thermal stability and is able to retain its fluorescence when fused to poorly folding proteins. We use a recently developed single-molecule technique which combines fluorescence-fluctuation spectroscopy and time-correlated single photon counting in order to characterize the photophysical properties of SFGFP under one- (OPE) and two- (TPE) photon excitation conditions. We use Rhodamine 110 as a model chromophore to validate the methodology and to explain the single-molecule results of SFGFP. Under OPE, single SFGFP molecules undergo fluorescence flickering on the time scale of micros and tens of micros due to triplet formation and ground-state protonation-deprotonation, respectively, as demonstrated by excitation intensity- and pH-dependent experiments. OPE single molecule fluorescence lifetimes indicate heterogeneity in the population of SFGFP, indicating the presence of the deprotonated I and B forms of the SFGFP chromophore. TPE of single SFGFP molecules results in the photoconversion of the chromophore. TPE of single SFGFP molecules show fluorescence flickering on the time scale of micros due to triplet formation. A flicker connected with protonation-deprotonation of the SFGFP chromophore is detected only at low pH. Our results show that SFGFP is a promising fusion reporter for intracellular applications using OPE and TPE microscopy. PMID- 16353268 TI - Evolution of subarachnoid space in normal fetuses using magnetic resonance imaging. AB - OBJECTIVE: The purpose of this study was to measure the fetal subarachnoid spaces at different sites of the brain using magnetic resonance (MR) images and analyze them in relation to gestational age. METHODS: Fetal MR images were obtained from 158 fetuses between 18 and 39 weeks of gestation who later showed no neurological problems. We bilaterally measured the distance between the superoanterior gyrus and the cranium as the frontal subarachnoid space (FSS) and the distance between the cortex in the parieto-occipital sulcus and the cranium as the parietal subarachnoid space (PSS). We also measured the cisterna magna between the cerebellar vermis and the cranium. Each of these was analyzed in relation to gestational age. RESULTS: The width of the FSS began to decrease significantly at 32 weeks of gestation (P < 0.05). The width of the PSS started to decrease significantly at 34 weeks of gestation (P < 0.05). There was no difference between the right and left sides (P < 0.05). The size of the cisterna magna showed a positive correlation with gestational age (P < 0.05). CONCLUSION: Measurement of the subarachnoid space is potentially useful for evaluating fetal development. PMID- 16353269 TI - Successful selective termination at 17 weeks' gestation in monochorionic monoamniotic twin pregnancy affected by twin-twin transfusion syndrome and discordant for hypoplastic left heart syndrome. AB - OBJECTIVES: To report a successful selective feticide in a complicated monochorionic monoamniotic (MCMA) pregnancy. METHODS: A case of MCMA pregnancy with severe twin-twin transfusion syndrome and discordant for hypoplastic left heart syndrome was diagnosed at 16 weeks' gestation. A complete ultrasound and fetoscopic surveillance was performed, ruling out cord entanglement and, thus, precluding the necessity of transecting the cord. RESULTS: The selective feticide was successfully performed by bipolar coagulation of the umbilical cord of the abnormal fetus under ultrasound guidance. The survivor twin developed normally during the rest of the pregnancy and was born at term. At 6 months of age, the infant was healthy. CONCLUSION: Selective feticide in complicated monoamniotic pregnancies can be safely performed. Cord entanglement can be confidently excluded by both ultrasound and fetoscopy, thus making the systematic transection of the umbilical cord unnecessary. PMID- 16353270 TI - Psychological consequences of termination of pregnancy for fetal anomaly: similarities and differences between partners. AB - OBJECTIVE: We examined the psychological responses to termination of pregnancy (TOP) for fetal anomaly from both men and women. The aim was to find risk factors for poor psychological outcome both for the individuals and for the couple. METHODS: A cross-sectional study was performed in 151 couples 2-7 years after TOP. We used standardized and validated questionnaires to investigate grief, symptoms of posttraumatic stress, somatic complaints, anxiety, and depression. RESULTS: Most couples adapted well to their loss, although several patients had pathological scores on posttraumatic stress symptoms and depression. Differences between men and women were slight. Higher education, good partner support, earlier gestational age, and life-incompatibility of the disorder positively influenced the outcomes, more for women than for men. Men and women with pathological scores rarely had such scores simultaneously. CONCLUSION: We emphasize the importance of equally involving both parents in the counselling because the outcomes of grief and posttraumatic stress symptoms between men and women only moderately differ and post-TOP psychopathology occurs in men as well. Good adjustment to TOP in women seems dependent on the level of support that they perceive from their partners. The intracouple results of the study suggest a mutual influence in the process of grieving between the partners. PMID- 16353271 TI - Intrauterine hemodynamic change in TTTS: the alterations in the inferior vena cava waveform in the donor fetus. AB - BACKGROUND: Significant hemodynamic changes are commonly observed in both fetuses in twin-twin transfusion syndrome. METHODS AND RESULTS: We describe two donor fetus cases in twin-twin transfusion syndrome associated with abnormal waveform patterns in the inferior vena cava after therapeutic amnioreduction. Subsequent echocardiographic examinations demonstrated progressive deterioration of the cardiac function in the recipient fetuses. CONCLUSIONS: We suggest that alterations in the inferior vena cava waveform in the donor fetuses may contribute to the accurate prediction of deterioration in fetal circulation after amnioreduction. PMID- 16353272 TI - Mild pyelectasis diagnosed by prenatal ultrasound is not a predictor of urinary tract morbidity in childhood. AB - OBJECTIVES: To determine whether children with prenatally diagnosed mild pyelectasis have more urinary tract morbidity during childhood than children without this finding. METHODS: Case-control study in children with pyelectasis (anteroposterior diameter of the fetal renal pelvis of 5-10 mm). A validated questionnaire was sent to the parents of 208 cases and 416 matched controls. RESULTS: The questionnaire was returned by 146 cases and 250 controls. There was a male predominance in the case group (p = < 0.0001). There was no difference in voiding habits or in prevalence of urinary tract infections (UTI). The prevalence of UTI was high: 11.6% in cases and 10.0% in controls. The only difference was a higher prevalence of constipation in the case group (p = 0.003). Postnatally, 41 children had an ultrasound examination and 16 were referred to a paediatrician or urologist: 3 had persisting pyelectasis, 3 had a recurrent UTI and 1 required surgery (a pyeloplasty). Four of the controls were referred to a paediatrician or urologist: 3 had recurrent UTI and 1 had urinary incontinence. CONCLUSION: Children with a mild fetal pyelectasis do not have more urinary tract morbidity during childhood than children without this finding. Therefore, there seems to be no need for additional investigation after birth. PMID- 16353273 TI - Expectation and knowledge of women undergoing first-trimester combined screening for Down syndrome in a Chinese population. AB - OBJECTIVES: To study the preference of pregnant women regarding the time taken to report the results of first-trimester combined screening for Down syndrome and their knowledge about it. METHODS: A questionnaire survey was conducted on a cohort of 325 pregnant participants who attended our first-trimester combined nuchal translucency and biochemical screening programme for fetal Down syndrome. This service was operated in a one-stop setting and the result of the screening test was available within 1 to 2 h after the collection of blood sample. RESULTS: The majority of participants: (1) could recall the quoted detection rate correctly (96.6%); (2) understood that a negative test does not exclude Down syndrome (91.1%); (3) understood that a positive test does not equate to an affected fetus (91.0%), and (4) could decide on the need for further invasive tests on the basis of the screening test results (98.1%). Twenty-nine percent of participants considered that a one-stop setting was very important because any delay in releasing the results made a significant difference to them, while 48.9% considered it acceptable if the results were available on the same day. The percentage dropped dramatically to 6.3% (within 2 days), 6.7% (within 3 days), 5.7% (within 1 week), and 3.1% (more than 1 week). CONCLUSION: The majority of the pregnant women in our test considered same day reporting of screening test results to be no different from a one-stop clinic. This would have important implications for the organisation and structuring of our service provision, since the operation of a one-stop clinic imposes significant stresses on the clinical staff involved. PMID- 16353274 TI - The influence of risk estimates obtained from maternal serum screening on amniocentesis rates. AB - OBJECTIVE: To investigate the influence of Down syndrome risk estimates obtained from maternal serum screening (MSS) on women's choices regarding amniocentesis. METHODS: Women who screened positive for Down syndrome by an Ontario MSS program between 1993 and 1998 were grouped on the basis of their risk estimate and ethnicity. Amniocentesis uptake rates between the groups were compared in order to determine how the MSS risk estimate influenced uptake. RESULTS: Analysis of 16 792 women showed that amniocentesis uptake rates increased as the estimated risk increased. Uptake in women < or = 35 was higher than that for older women (70% vs 60%, p = 0.001). Uptake in Caucasian and Asian women was higher than the uptake in Black women (67% vs 49%, p = 0.001). Women aged 35 years or older were more likely to proceed with amniocentesis if the MSS risk estimate was higher than their age-specific risk. CONCLUSION: The increase in amniocentesis rate paralleled the increase in MSS risk estimate for Down syndrome. Risk-specific amniocentesis rates are higher in women aged less than 35 years. Women aged 35 years or older whose risk estimate by MSS is lower than their age-specific risk are less likely to opt for amniocentesis. PMID- 16353275 TI - The effect of fetal gender on the false-positive rate of Down syndrome by maternal serum screening. AB - OBJECTIVES: (1) To further explore if there is a difference in maternal serum levels of alpha-fetoprotein (AFP), human chorionic gonadotrophin (hCG) and estriol (uE3) between fetal genders. (2) To determine if these differences influence false-positive rates of Down syndrome screening in pregnancies with male or female fetuses. METHODS: This is a descriptive study of women screened at the Ontario Maternal Serum Screening program between 1993 and 1995. The women were grouped by fetal gender and ethnicity. Serum levels of the three markers and screening false-positive rates for Down syndrome were compared between fetal genders in women of different ethnicity respectively. RESULTS: Complete data were available for 110 306 pregnancies. In all three ethnic groups, MSAFP levels were significantly decreased and MShCG levels were significantly increased in women with female fetuses. The level of MSuE3 was similar between genders. The difference in false-positive rates of Down syndrome between genders was not statistically significant. CONCLUSIONS: This is the largest study comparing false positive rates between fetal genders. In contrast to previous studies, the differences in the serum marker levels between fetal genders do not influence the false-positive rates for Down syndrome. PMID- 16353276 TI - Synthesis and antiviral evaluation of novel 4'-hydroxymethyl branched thioapiosyl nucleosides. AB - In this study, we synthesized novel 4'-hydroxymethyl branched thioapiosyl nucleosides. The thioapiosyl sugar moiety was constructed using sequential ozonolysis and reduction. The natural bases (uracil, thymine, cytosine, and adenine) were efficiently coupled using the Vorbruggen glycosyl condensation procedure (persilyated base and TMSOTf). The antiviral activities of the synthesized compounds were evaluated against HIV-1, HSV-1, HSV-2, and HCMV. Compound 19 showed moderate anti-HIV activity (EC(50) = 19.3 microg/mL) without exhibiting any cytotoxicity up to 100 microM. PMID- 16353277 TI - New synthetic thiocolchicine derivatives as lowtoxic anticancer agents. AB - New thiocolchicine derivatives (1-8) were designed as less toxic anticancer agents possessing the power full anticancer activity of colchicine. The synthesis and biological evaluation of these compounds were described. As a preliminary result of biological in vitro investigation, compounds 1, 6, and 7 showed lower toxicities than that of colchicine in combination with potent anticancer activities. PMID- 16353278 TI - Cyclooxygenase-1 and cyclooxygenase-2 inhibition of novel 1,2-disubstituted imidazoles. AB - Novel 1,2-disubstituted imidazoles were synthesized and tested for COX-1 and COX 2 inhibition. A substituted phenyl cycle in position 1 and an alicyclic tertiary alcohol in position 2 gave COX-1-selective compounds with potency and selectivity depending on the different substituents. PMID- 16353281 TI - Role of fetal ultrasound and magnetic resonance imaging in the prenatal diagnosis of migration disorders. AB - OBJECTIVES: To evaluate the contribution and limitations of fetal ultrasonography (US) and magnetic resonance imaging (MRI) for the diagnosis and management of migration disorders. METHODS: Over a 5-year period, 14 fetuses with pathological migration disorders, without an infectious context, were taken care of in our centre. All underwent US; nine underwent MRI as well. Sonographic and MRI data were compared with neuropathological data. RESULTS: The diagnosis of gyral disorders was obtained by US in 1/14 cases; other cerebral abnormalities were found suggesting neuronal disorder in the remainder. Cerebral MRI suggested gyral abnormality in eight of the nine cases. CONCLUSIONS: US performance is increasing. MRI appears to be a promising method for the diagnosis of fetal migration disorders, giving better results than US. It may be recommended in cases of abnormal cerebral US findings or familial clinical history. However, interpretation of MRI can be tricky and the resulting diagnosis occurs late within the pregnancy. PMID- 16353282 TI - Prenatal diagnosis of PLP1 copy number by array comparative genomic hybridization. AB - OBJECTIVES: To report a family with a history of Pelizaeus-Merzbacher disease (PMD) for which prenatal diagnosis of PLP1 gene duplication status was attempted by the use of custom array comparative genomic hybridization (aCGH). METHODS: A 28-year-old woman was referred for genetic counseling for her then current pregnancy because her existing 3-year-old son was diagnosed with a classic form of PMD. At 11 and 3/7 weeks gestation, chorionic villus sampling (CVS) was performed. Custom aCGH and fluorescence in situ hybridization (FISH) analyses were also performed on the DNA from family members. Fetal karyotyping revealed 46,XY. RESULTS: Analysis by aCGH revealed that the male fetus was not duplicated for the PLP1 gene, but confirmed a duplicated PLP1 gene in the 3-year-old son, and that the mother was a duplication carrier. These results were independently confirmed by FISH analysis. aCGH and FISH analyses on DNA and cells derived from cord blood confirmed PLP1 nonduplication in the newborn. CONCLUSION: aCGH is a reliable alternative method for detection of PLP1 copy number for prenatal diagnosis of Pelizaeus-Merzbacher disease. PMID- 16353283 TI - A comparison of prenatal versus postnatal karyotyping for the investigation of intrauterine fetal death after the first trimester of pregnancy. AB - INTRODUCTION: Aneuploidy is an important cause of intrauterine fetal death (IUFD) after the first trimester. Determination of the fetal karyotype of these pregnancies is commonly done in most units from solid tissues. Results from such techniques are disappointing. The aim of this audit was to compare the results of karyotyping IUFD by invasive testing (amniocentesis or chorionic villus sampling) and solid tissues (skin biopsy). SUBJECTS AND METHODS: Women with IUFD managed in our unit between 1 January 1998 and 31 December 2002 (inclusive) were offered either invasive testing before medical induction of labour or solid tissue biopsy after delivery. The amniotic fluid, chorionic villi and biopsies were processed following standard laboratory procedures. RESULTS: During the 60 months, 230 samples from cases of IUFD were received by the laboratory in our unit; 126 had skin biopsies and 104 underwent invasive testing (81 amniocenteses and 23 chorionic villus sampling). Successful karyotyping was possible in 90% of those who underwent amniocentesis, 100% of those who had chorionic villus sampling and 13.5% of those who had skin biopsies. 50% of skin biopsies were unsuitable for analysis compared to none in the CVS and amniocentesis group. The difference in successful karyotyping between invasive testing and solid tissue testing was statistically significant (P < 0.0001). There were 12 (10.6%) abnormal karyotypes from the 113 successful samples (11/96 in the invasive group versus 1/17 in the solid tissue group). CONCLUSION: Invasive testing has a much higher success rate of karyotyping in cases of IUFD and should, therefore, be offered to women presenting with this complication irrespective of gestational age. PMID- 16353284 TI - Methylenetetrahydrofolate reductase gene polymorphisms and their association with trisomy 21. AB - OBJECTIVES: To verify whether the frequencies of 5,10-methylenotetrahydrofolate reductase (MTHFR) polymorphisms at positions 677 and 1298 are higher in women with children affected by trisomy 21 than in those with chromosomally normal offspring. METHODS: A case-control study was carried out with 70 women whose children had trisomy 21 and 88 controls whose children were chromosomally normal. The frequencies of polymorphisms of points C677T and A1298C of MTHFR gene coding were studied in these two groups. Odds ratios (OR) for having a child affected by trisomy 21 were estimated for homozygosis, heterozygosis or the absence of the above-mentioned MTHFR polymorphisms. Logistic regression models were used to control for the effect of confounding variables on these odds ratios. RESULTS: The frequency of joint heterozygotic polymorphism (677 and 1298) was significantly higher in women with children affected by trisomy 21 than in those with chromosomally normal offspring (OR: 5.7). CONCLUSIONS: The presence of joint heterozygotic polymorphism in the codifying gene for MTHFR was a risk factor for having a child with trisomy 21. PMID- 16353285 TI - Preimplantation genetic diagnosis (PGD) for Duchenne muscular dystrophy (DMD) by triplex-nested PCR. AB - OBJECTIVES: Duchenne muscular dystrophy (DMD) is a lethal X-linked recessive disorder with an incidence of approximately 1 in 3500 males, caused by mutation in the DMD gene. About 2/3 of DMD cases are caused by gross DMD gene deletion mutations. The purpose of this study was to develop a series of single-cell multiplex-nested PCR protocols for preimplantation genetic diagnosis (PGD) of the most prevalent DMD deletions. METHODS: The protocols were developed on single blood leukocytes from normal males and females and patients with known DMD gene deletion. In the first reaction, 2 of 11 different primer sets (exons 4, 8, 12, 13, 17, 46, 47, 49, 50, 52 and intron 52) were used to allow the simultaneous amplification of different DMD loci and the SRY gender marker, in a single triplex-nested polymerase chain reaction (PCR). Aliquots of this reaction were then subjected to nested PCR in which each locus was amplified individually. Following the successful establishment of single-cell triplex-nested PCR in single leukocytes, the technique was employed in five clinical PGD cases. RESULTS: For each DMD locus, more than 50 single leukocytes from healthy controls and more than 100 single leukocytes from affected individuals with known deletions were analyzed. Amplification efficiency for each tested locus was 98 100%. The false-negative rates for each analysis taken separately was <1%. Taken together, however, the results of the triplex-nested PCR analysis had a false negative rate of 0%. No contamination was detected in all wash-drop blanks tested. We subsequently performed 18 PGD cycles in 5 DMD carriers. A total of 156 embryos were biopsied and successfully analyzed. Of these, 39 affected embryos were detected and 50 unaffected embryos were transferred (mean = 2.9 +/- 1.1 embryos per cycle). These resulted in three biochemical pregnancies and three clinical pregnancies, all of which have culminated in the birth of normal offspring. CONCLUSION: Triplex-nested PCR using 2 of 11 DMD loci and the SRY gender marker allow PGD for >90% of DMD families with known deletions. These protocols are associated with a high amplification efficiency and accuracy. PMID- 16353286 TI - Rapid prenatal diagnostic testing for Down syndrome only or longer wait for full karyotype: the views of pregnant women. AB - OBJECTIVES: Prenatal diagnosis by karyotype analysis determines the copy number and structure of each chromosome and is considered to be the 'gold standard' in detection of chromosome abnormality. This method is, however, time consuming and women may have to wait up to 21 days for the results. With improving molecular genetic techniques, the most commonly occurring chromosome abnormalities can be diagnosed within 2 days from amniotic fluid. This study investigates the value pregnant women place on these alternative prenatal diagnostic tests. METHODS: A structured self-completion discrete choice experiment questionnaire was designed. The subjects were 40 pregnant women, of unknown risk, attending for their 11-week booking scan and 10 elevated-risk women attending for amniocentesis at the Aberdeen Maternity Hospital, Scotland. RESULTS: Women value 'comprehensive' information of a full karyotype analysis at pounds 791, and 'simple' information of a rapid diagnostic test at pounds 690. Willingness to pay for a day's reduction in waiting time for results is pounds 18. Women prefer a prenatal service providing simple over comprehensive information as long as they receive results 6 days sooner than the comprehensive results would be provided. CONCLUSIONS: While women prefer comprehensive information, the bulk of the value is placed on knowing whether the fetus has Down syndrome. Given the longer wait times for comprehensive information, simple information is preferred as long as results are received 6 days sooner than would be the case for comprehensive information. These results have implications for the resources dedicated to providing a rapid prenatal diagnostic service. PMID- 16353287 TI - Ductus venosus Doppler velocimetry in the prediction of acidemia at birth: which is the best parameter? AB - OBJECTIVES: To evaluate the prediction of acidemia at birth using ductus venosus Doppler velocimetry and to determine the best parameter and cut-off values for this prediction in pregnancies complicated with placental insufficiency. METHODS: Prospective cross-sectional study. Forty-seven patients with placental insufficiency managed in two Brazilian hospitals were submitted to ductus venosus Doppler velocimetry in the last 24 h before delivery. All pregnancies were singleton, at least 26 weeks of age and without structural or chromosomal anomalies. A ROC curve was calculated for each ductus venosus parameter (independent variable) and acidemia (dependent variable). A cut-off value was established. The McNemar test was used to compare the various parameters. RESULTS: The ductus venosus S, D and A peak velocities were not good predictors of acidemia at birth. Pulsatility Index for Veins (PIV) was a good predictor of acidemia (ROC curve area 0.79, p = 0.003), as well as S/A and (S - A)/S ratios (ROC curve area 0.818, p = 0.001). The cut-off values were PIV = 0.76, S/A = 2.67 and (S - A)/S = 0.63. CONCLUSIONS: In this high-risk population, angle independent ductus venosus Doppler indexes were good predictors of birth acidemia. The S/A and (S - A)/S ratios and the ductus venosus PIV were statistically equivalent in this prediction. PMID- 16353288 TI - Rasch analysis of the dimensional structure of the Hospital Anxiety and Depression Scale. AB - The Hospital Anxiety and Depression Scale (HADS) has been used extensively in cancer patients to identify psychological distress. Reports of the factor structure and screening performance of the instrument vary. Rasch models allow an assessment of the structure of a questionnaire by identifying item fit. Removal of misfitting items may improve both the dimensionality and efficacy of screening questionnaires. A Rasch analysis of the HADS-T and subscales was used to explore the factor structure, dimensionality and screening efficacy. A total of 1855 patients completed a touchscreen version of the HADS, including 381 patients who had received a psychiatric interview (SCAN/PSE). These data were analysed using Rasch models, and the screening efficacy at identifying cases of psychological distress and anxiety and depression evaluated. The results demonstrated that the structure of the HADS-T and subscales was unidimensional. Three items from the HADS-T, and one from each of the subscales demonstrated misfit. Screening efficacy for the HADS-T and subscales was modest. However, removal of misfitting items had little impact on screening, demonstrating that items could potentially be omitted, if required. The item range covered a narrow spectrum of psychological distress, predominantly higher levels of distress. Additional items have to be added if screening for moderate to mild distress is to be improved for cancer patients. PMID- 16353289 TI - Mechanism of in vivo DNA transport into cells by electroporation: electrophoresis across the plasma membrane may not be involved. AB - BACKGROUND: Recently, in vivo gene transfer with electroporation (electro-gene transfer) has emerged as a leading technology for developing nonviral gene therapies and nucleic acid vaccines. The widely hypothesized mechanism is that electroporation induces structural defects in the membrane and provides an electrophoretic force to facilitate DNA crossing the permeabilized membrane. In this study, we have designed a device and experiments to test the hypothesis. METHODS: In this study, we have designed a device that alternates the polarity of the applied electric field to elucidate the mechanism of in vivo electro-gene transfer. We also designed experiments to challenge the theory that the low voltage (LV) pulses cannot permeabilize the membrane and are only involved in DNA electrophoresis, and answer the arguments that (1) the reversed polarity pulses can cause opposing sides of the cell membrane to become permeabilized and provide the electrophoresis for DNA entry; or (2) once DNA enters cytoplasmic/endosomal compartments after electroporation, it may bind to cellular entities and might not be reversibly extracted. Thus a gradual buildup of the DNA in the cell still seems quite possible even under the condition of the rapid reversal of polarity. RESULTS: Our results indicate that electrophoresis does not play an important role in in vivo electro-gene transfer. CONCLUSIONS: This study provides new insights into the mechanism of electro-gene transfer, and may allow the definition of newer and more efficient conditions for in vivo electroporation. PMID- 16353290 TI - Matrix-assisted laser desorption/ionization of peptides on AnchorChip targets with alpha-cyano-4-hydroxycinnamic acid and nitrocellulose as matrix. PMID- 16353291 TI - Investigation of interconversion between aspacochiosides A and B by fast-atom bombardment mass spectrometry. PMID- 16353292 TI - Cluster ion formation between 2,5-diphenyl-1,3,4-oxa(thia)diazole and alkali earth metal cations studied by electrospray ionization mass spectrometry. PMID- 16353294 TI - Winning against heart disease, losing against diabetes. PMID- 16353293 TI - Cytokine gene expression profile distinguishes CD4+/CD57+ T cells of the nodular lymphocyte predominance type of Hodgkin's lymphoma from their tonsillar counterparts. AB - Little is known about the cytokine profile of nodular lymphocyte predominance Hodgkin's lymphoma (NLPHL) and the significance of the characteristic rosetting CD4(+)/CD57(+) T cells. We analysed the T lymphocyte populations isolated from lymph node suspensions from five patients with NLPHL, two with follicular hyperplasia and progressive transformation of germinal centres (PTGC), three with classical Hodgkin's lymphoma (CHL) and five with hyperplasia of the tonsil. We sorted the T cells based on expression of CD3, CD4 and CD57 by flow cytometry and evaluated the cytokine mRNA profiles of the T cells with quantitative RT-PCR. NLPHL cases were as rich in T cells as the CHL cases, but all NLPHL cases had a much higher frequency of CD4(+)/CD57(+) T cells. In contrast to the CD4(+)/CD57(+) T cells from tonsils, IL2 and IL4 mRNAs were consistently absent from the CD4(+)/CD57(+) T cells of NLPHL. Even after stimulation, no IL4 transcripts could be detected in the CD4(+)/CD57(+) T cells of NLPHL. On the other hand, IFNgamma transcripts were elevated in NLPHL and PTGC T cell subsets as compared to tonsillar T cell subsets. IL13 mRNA was exclusively produced by the T cells of CHL cases, indicating that IL13 may be a key cytokine in CHL. In conclusion, elevated levels of CD4(+)/CD57(+) T cells are characteristic of NLPHL and these T cells display a distinct cytokine mRNA profile. PMID- 16353295 TI - Lower blood pressure may provide dementia protection. PMID- 16353296 TI - Years ago, I saw a TV medical show in which a doctor asked a patient to raise his arms over his head and report on whether his chest pain got better or worse. I've noticed that sometimes I have sharp pains in my chest, on one side or the other, particularly after I've carried suitcases. The pain often gets worse when I raise my arms. Does that indicate a heart condition or just muscle strain? PMID- 16353297 TI - I seem to be losing height as I age. In calculating body mass index, should I use current height or previous mature adult height in doing the math? Using my current height, my BMI is 28.2. But my previous mature adult height gives me a BMI of 25.8--a major difference. PMID- 16353298 TI - Presidential address. The nexus: where science meets society. PMID- 16353299 TI - Fluid solutions in dengue shock syndrome. PMID- 16353300 TI - Dengue in travelers. PMID- 16353301 TI - Dengue in travelers. PMID- 16353302 TI - Perioperative beta-blocker therapy and mortality. PMID- 16353303 TI - Perioperative beta-blocker therapy and mortality. PMID- 16353304 TI - Abstracts of the 18th European College of Neuropsychopharmacology Congress, Amsterdam, The Netherlands, October 22-26, 2005. PMID- 16353305 TI - Phthalate linked to lupus in mice. PMID- 16353306 TI - New human retroviruses. PMID- 16353307 TI - Beyond the bench: nurses adapt to changing health care climate. PMID- 16353308 TI - HIV vaccine moves to phase II. PMID- 16353309 TI - Avian influenza. PMID- 16353310 TI - Russia clamps down on foreign aid organisations. PMID- 16353311 TI - Malawi's food crisis continues to worsen. PMID- 16353312 TI - Controversial appointment upsets Czech health ministry. PMID- 16353313 TI - Combination chemotherapy with adriamycin, cyclophosphamide, vincristine, methotrexate, etoposide and dexamethasone (ACOMED) followed by involved field radiotherapy induces high remission rates and durable long-term survival in patients with aggressive malignant non-Hodgkin's lymphomas: long-term follow-up of a pilot study. AB - The aim of the present study was to evaluate the feasibility and efficacy of the intensified induction chemotherapy regimen ACOMED for patients with aggressive non-Hodgkin's lymphoma (NHL). Untreated adult patients with aggressive NHL, presenting with Ann Arbour stage II-IV disease or stage I with bulky disease, and with at least one of the following risk factors: age > 60 years, advanced disease, elevated serum lactate dehydrogenase level, Eastern Cooperative Oncology Group (ECOG) performance status >or= 2, presence of extranodal sites of disease and bulky disease, were treated with the ACOMED regimen consisting of 4-6 cycles of adriamycin 25 mg/m(2) i.v. on days 4-5, cyclophosphamide 250 mg/m(2) i.v. on days 1-5, vincristine 2 mg i.v. absolute on day 1, methotrexate 500 mg/m(2) i.v. on day 1 with leucovorin-rescue after 24 h 30 mg/m(2) i.v. and 3 x 15 mg p.o., etoposide 100 mg/m(2) i.v. on days 3-5, dexamethasone 10 mg/m(2) p.o. on days 1-5 and granulocyte colony-stimulating factor support, repeated on day 21. Twenty-two patients were treated within this study at a single center. After 4-6 cycles of ACOMED followed by additional involved field radiotherapy in 18 patients, the complete and overall response rates were 86% (19 of 22 patients) and 95% (21 of 22 patients), respectively. After a median observation time of 10 years and 2 months, 16/22 (73%) patients are alive in continuous complete response without evidence of any late toxicities. ACOMED followed by involved field radiation presents a highly effective regimen for remission induction and long-term survival in patients with aggressive NHL, and merits further investigation. PMID- 16353314 TI - Posterior leukoencephalopathy after combination chemotherapy in a patient with lymphoma. AB - Posterior leukoencephalopathy syndrome is a recently described syndrome involving mainly parieto-occipital gray/white matter of the brain. It occurs secondary to various clinical entities, like hypertension and immunosuppressive therapy. Few cases after combination chemotherapy have been reported. This study describes a 36-year-old woman with primary refractory T-cell lymphoma, who developed central nervous system toxicity due to treatment with intrathecal methotrexate and intravenous ifosfamide, idarubicine and etoposide given as a salvage regimen. Both clinical features as well as magnetic resonance imaging findings were typical for posterior leukoencephalopathy syndrome. The patient died despite anti hypertensive therapy and haemodialysis. Central nervous system toxicity related to chemotherapeutics and posterior leukoencephalopathy syndrome are discussed briefly. PMID- 16353315 TI - Modulation of the specific interaction of cardiolipin with Cytochrome c by Zwitterionic phospholipids in binary mixed bilayers: a 2H and 31P-NMR study. PMID- 16353317 TI - Surgeons pleased with patient's progress after face transplant. PMID- 16353318 TI - In celebration of cerebration. PMID- 16353319 TI - Bleeding profile and endometrial safety of continuous combined regimens 1 mg 17beta-estradiol/trimegestone versus 1 or 2 mg 17beta-estradiol/norethisterone acetate in postmenopausal women. AB - OBJECTIVE: To compare the bleeding profile and endometrial safety of continuous combined 1 mg 17beta-estradiol (17beta-E2) and 0.125 mg trimegestone (TMG) with those of two continuous combined 17beta-E2 and norethisterone acetate (NETA) regimens. STUDY DESIGN: This was a double-blind, randomized, multicenter study conducted in 12 European countries and Israel over a 2-year period. Healthy postmenopausal women with an intact uterus were given either 1 mg 17beta-E2/0.125 mg TMG, 2 mg 17beta-E2/1 mg NETA or 1 mg 17beta-E2/0.5 mg NETA for up to 26 cycles, each of 28 days. RESULTS: The percentage of amenorrheic women was greater in most cycles up to cycle 13 in the 1 mg 17beta-E2/0.125 mg TMG group than in the comparator groups. The mean number of bleeding days was similar in the 1 mg 17beta-E2/0.125 mg TMG and the 1 mg 17beta-E2/0.5 mg NETA groups, but greater in the 2 mg 17beta-E2/1 mg NETA group. No endometrial hyperplasia was observed for any group. CONCLUSION: Continuous combined 1 mg 17beta-E2/0.125 mg TMG exhibits a more favorable bleeding profile than 1 mg 17beta-E2/0.5 mg NETA up to 1 year, while providing an adequate protective effect on the endometrium. PMID- 16353321 TI - Sub-Saharan region remains hotspot as disease spreads. PMID- 16353320 TI - Reasons for exclusion from thrombolytic therapy following acute ischemic stroke. PMID- 16353322 TI - The doctor-patient relationship: a power shift to patients. PMID- 16353323 TI - Prevention: science with your fruit and vegetables. PMID- 16353324 TI - Cancer: battles won, but the war continues. PMID- 16353326 TI - Obesity: heavyweight champions of the world. PMID- 16353325 TI - Genetics begets genomics: tailor-made medicine. PMID- 16353327 TI - Mental health: Prozac changes moods...and minds. PMID- 16353328 TI - Replacement parts: organ transplants and stem cells. PMID- 16353329 TI - From the founding editors. Beginners' luck, pluck and conviction. PMID- 16353330 TI - [The significance of sickle cell anemia within the context of the Brazilian government's 'racial policies' (1995-2004)]. AB - This essay reflects on the social significance of growing interest in sickle cell anemia and other illnesses associated with the black body in Brazil. I explore the discursive network that has taken shape around the disease within the social context of its production. I first summarize anthropologist Melbourne Tapper's analysis of the United States program to fight sickle cell anemia in the 1970s, shortly after blacks attained victories in the civil rights movement. Tapper (1999) argues that one of the consequences of this policy was the creation of a responsible black citizenry. In the late 1990s, the Brazilian government developed a program (Programa de Anemia Falciforme) that counted on the heavy participation of black activists and that also contributed to the formation of a "responsible black community". My argument is that sickle cell anemia becomes much more than an illness to be eradicated. The discourse surrounding it is a powerful element in the process of naturalization of the "black race" (and, by logical and political complement, the "white race") in a country that until recently imagined itself a biologically and culturally hybrid nation. PMID- 16353331 TI - Can ethnic differences in men's preferences for women's body shapes contribute to ethnic differences in female adiposity? AB - In the United States, obesity is more common among black and Hispanic than white women. One putative cause of this difference is different cultural norms for attractiveness. Two studies assessed ethnic differences in men's perceptions of the attractiveness of females of varying sizes. In the first, 108 men recruited on the New York subway were shown sets of silhouettes depicting female bodies varying in fatness and were asked to pick the silhouette they found most attractive. They were also asked to indicate the thinnest and fattest figures they would consider dating. A measure of "latitude of acceptance" was computed as the difference between the thinnest and fattest figures considered. Results indicated no relationship between ethnicity and preference (F = 1.383, p = .257) or "latitude" (F = .102, p = .903). In Study 2, "personal advertisements" placed by 373 black, 1915 white, 110 Hispanic, and 30 Asian men from 35 newspapers and magazines were coded as: 1) thinness preferred; 2) no information on weight preference; 3) fatness preferred; or 4) states weight or looks unimportant. Results indicated a statistically significant but small association between ethnicity and preference (chi2 = 49.55, df = 9, p < .00001). Relative to white and Asian men, black and Hispanic men more frequently requested fat women, Hispanic men less frequently requested thin women, and black men more frequently stated that looks or weight did not matter. Ethnicity explained only 2.1% of the variance in preference. Thus, it seems unlikely that ethnic differences in men's preferences for women's body shapes contribute substantially to ethnic differences in female adiposity. PMID- 16353332 TI - Lipolysis in intraabdominal adipose tissues of obese women and men. AB - Intraabdominal fat in humans is located in two major depots, the omental and mesenteric. We compared basal and stimulated lipolysis in adipose tissue from these two depots and the subcutaneous abdominal depot of obese women and men. Omental fat cells of women are smaller and have lower rates of basal lipolysis than in men. Basal lipolysis rates are significantly higher in subcutaneous than intraabdominal adipose tissues of both genders. In men, the incremental lipolytic response to norepinephrine is significantly greater in both intraabdominal fat depots than in the subcutaneous fat, while in women the response of the mesenteric is lower than the omental. In women, but not men, responsiveness to the beta agonist isoproterenol is also increased in omental tissue. Thus, in women, omental and mesenteric adipose tissues show distinctly different metabolic properties which may moderate the impact of intraabdominal obesity. PMID- 16353333 TI - Insulin secretion in hypothalamic obesity: diurnal variation and the effect of naloxone. AB - This paper has tested the hypothesis that patients with hypothalamic obesity have altered mechanisms controlling insulin secretion when compared to obese patients without hypothalamic injury. Fasting glucose and insulin values were significantly higher in the morning than in the afternoon in the six control obese patients, but there was no diurnal difference in the six patients with hypothalamic obesity (n=6). The control obese subjects showed a diurnal variation in glucose-stimulated insulin secretion, whereas the patients with hypothalamic obesity did not, suggesting that hypothalamic injury had destroyed diurnal rhythms. Naloxone, an opioid antagonist, acutely suppressed fasting insulin in the six patients with essential obesity but had little effect on fasting insulin in the three patients with hypothalamic obesity or in five normal-weight controls. Naloxone increased insulin sensitivity in the obese control patients, but did not affect either insulin secretion or insulin sensitivity in patients with hypothalamic obesity or in normal weight subjects. Our results support the conclusion that hypothalamic obesity disrupts diurnal rhythms, with the suggestion that opioid peptides affect insulin secretion differently in patients with essential obesity as compared to normal weight subjects or those with hypothalamic obesity. PMID- 16353334 TI - Insulin action in rats is influenced by amount and composition of dietary fat. AB - The chronic influence of dietary fat composition on obesity and insulin action is not well understood. We examined the effect of amount (20% vs 60% of total calories) and type (saturated vs polyunsaturated) of fat on insulin action and body composition in mature male rats. Six months of feeding a high fat (HF) diet led to obesity and impaired insulin action (determined by a euglycemic hyperinsulinemic clamp), neither of which were reversed by a subsequent 6 months of feeding a low fat (LF) diet. Within HF fed rats, type of fat did not affect body composition or insulin action. Six months of feeding a low fat diet led to only a slight decline in insulin action, with no difference due to type of dietary fat. From 6-9 months, insulin action became more impaired in LF rats fed the saturated diet than in LF rats fed the polyunsaturated diet. By 12 months, all groups were obese and had a similar impairment in insulin action. The amount and type of fat in the diet did not influence the overall degree of impairment in insulin action but did affect the time course. Both feeding a high fat diet and feeding a low fat saturated diet accelerated the impairment in insulin action relative to rats fed a low fat polyunsaturated fat diet. PMID- 16353335 TI - Using an immortalized cell line to study the HPV life cycle in organotypic "raft" cultures. AB - The papillomavirus life cycle is tied to the differentiation of the stratified squamous epithelium that this virus infects. The ability to study the papillomavirus life cycle is facilitated by organotypic culturing techniques that allow one to closely recapitulate this terminal differentiation process in the laboratory. Current techniques allow for the establishment of recombinant wild type or mutant human papillomavirus (HPV) genomes in transfected early-passage human foreskin keratinocytes (HFKs). These cells can then be used in organotypic culture to investigate the role of individual viral genes in different aspects of the viral life cycle. When using early-passage HFKs, there is a need for the transfected HPV genome to extend the life span of the cells in order to have sufficient cell generations in which to carry out organotypic culturing. The recent isolation of a spontaneously immortalized HFK cell line that supports the complete HPV life cycle has further allowed investigators to study wild-type or mutant papillomaviral genomes that do not confer immortalization. In this chapter, we describe the methodologies that permit the study of the HPV life cycle in this HFK cell line. PMID- 16353336 TI - Retrovirus-mediated gene transfer to analyze HPV gene regulation and protein functions in organotypic "raft" cultures. AB - The productive phase of human papillomavirus (HPV) infection is dependent on squamous differentiation of epithelial keratinocytes. Organotypic culture systems of primary human keratinocytes (PHKs) or immortalized keratinocytes that contain HPV genomes were developed to recapitulate this permissive environment. A complementary approach to determine the functions of individual HPV genes and to examine the virus-host interactions is to introduce the gene, alone or in combination, into keratinocytes that are then grown in organotypic cultures. The success of the latter approach depends on the methodology of retrovirus-mediated gene transfer, which can transduce the viral gene or genes into an entire population of PHKs. In this chapter, we describe the strategies and methods of retrovirus-mediated gene transfer into keratinocytes grown into organotypic cultures. PMID- 16353337 TI - The HPV xenograft severe combined immunodeficiency mouse model. AB - The permissive propagation of papillomaviruses outside their natural hosts has not been possible, which is an important restriction to the study of human papillomavirus (HPV) infections and associated diseases. Since the mid-1980s, several models have been described that rely on the growth of HPV in susceptible human xenografts implanted in immunodeficient mice. The severe combined immunodeficiency (SCID) mouse has been particularly suited to this approach, and to reproduce reliably the macroscopic, microscopic, and molecular features of naturally occurring lesions. We describe two common methods that permit growth and propagation of HPV in subcutaneous (heterotopic) or cutaneous (orthotopic) human skin grafts implanted in the SCID mouse. PMID- 16353338 TI - Replication and encapsidation of papillomaviruses in Saccharomyces cerevisiae. AB - Improvements in methodologies to recapitulate and study particular biological functions of the papillomavirus life cycle have led to great advances in our knowledge of these viruses. Described in this chapter are techniques that allow low-copy and high-copy replication of full-length human papillomavirus (HPV) genomes, as well as assembly of virus-like particles, in Saccharomyces cerevisiae (yeast). This system has several distinct advantages that make it an attractive complement to the well-established raft-culturing system. First, yeast are inexpensive, rapid, and simple to culture in the lab. Second, they provide an ever-widening array of genetic tools to analyze HPV functions--most recently notable, the yeast open reading frame (ORF)-deletion library. Third, yeast provide a potentially high-efficiency means to produce large quantities of infectious virus in a short time frame. Fourth, assembly of HPV virus in yeast allows encapsidation of mutant genomes, since previous studies have shown that no viral ORF is required for replication of full-length HPV in yeast. PMID- 16353339 TI - Analysis of the regulation of viral transcription. AB - Despite the small genomes and number of genes of papillomaviruses, regulation of their transcription is very complex and governed by numerous transcription factors, cis-responsive elements, and epigenetic phenomena. This chapter describes the strategies of how one can approach a systematic analysis of these factors, elements, and mechanisms. From the numerous different techniques useful for studying transcription, we describe in detail three selected protocols of approaches that have been relevant in shaping our knowledge of human papillomavirus transcription. These are DNAse I protection ("footprinting") for location of transcription-factor binding sites, electrophoretic mobility shifts ("gelshifts") for analysis of bound transcription factors, and bisulfite sequencing for analysis of DNA methylation as a prerequisite for epigenetic transcriptional regulation. PMID- 16353340 TI - Analysis of HPV DNA replication using transient transfection and cell-free assays. AB - The genomes of human and animal papillomaviruses amplify in keratinocytes undergoing terminal squamous differentiation. Two approaches have been developed to facilitate the investigation into the requirement for viral DNA replication and its regulation outside the context of the host tissues. Under these conditions, the investigation can be conducted independently of viral genes required to re-establish the S-phase milieu in the differentiated keratinocytes to support viral DNA replication. The first method is transient replication in cell lines after transfection of a plasmid containing the viral origin of replication together with expression vectors of the necessary viral proteins that direct specific initiation from the origin. The second is cell-free replication in which purified viral replication proteins are complemented by cell extracts to initiate origin-specific replication. The two methods have identified the origin, the viral E1 and E2 proteins necessary for initiation, their functions, and the host factors that are required to support and regulate the viral DNA replication. These two methods are complementary in providing answers and insights that either one alone may not be able to achieve. This chapter provides a practical guide to these two replication assays. PMID- 16353342 TI - [Sex, hormones and Alzheimer's disease]. PMID- 16353341 TI - The development of retinotectal maps: a review of models based on molecular gradients. AB - Information about the world is often represented in the brain in the form of topographic maps. A paradigm example is the topographic representation of the visual world in the optic tectum/superior colliculus. This map initially forms during neural development using activity-independent molecular cues, most notably some type of chemospecific matching between molecular gradients in the retina and corresponding gradients in the tectum/superior colliculus. Exactly how this process might work has been studied both experimentally and theoretically for several decades. This review discusses the experimental data briefly, and then in more detail the theoretical models proposed. The principal conclusions are that (1) theoretical models have helped clarify several important ideas in the field, (2) earlier models were often more sophisticated than more recent models, and (3) substantial revisions to current modelling approaches are probably required to account for more than isolated subsets of the experimental data. PMID- 16353343 TI - Exhaled breath analysis: from occupational to respiratory medicine. AB - Breath analysis is a technique rapidly gaining ground as a non-invasive tool to diagnose and monitor various aspects of lung diseases. Measurement of exhaled breath is safe, rapid, simple to perform, and effort independent. Given that human breath contains upwards of 250 chemicals, the potential for developing new applications is high. Much of the current knowledge on breath analysis in respiratory medicine derives from years of experience gained in occupational settings, where breath analysis has been used mainly to assess exposure to volatile chemicals. Laboratory based analysis of exhaled air is a complex, expensive and time consuming process and thus is not in wide spread use in occupational medicine. However, recent knowledge of exhaled breath analysis in pulmonology, in particular in bronchial asthma and lung cancer, and the development of fast, and easy to perform non-invasive procedures for breath analysis, re-opened possible application of exhaled breath as a novel approach for biological monitoring of inhaled pneumotoxic substances. The simultaneous quantification of biomarkers of dose and effect in exhaled air may provide new insights into lung damage occurring in workers exposed to inhaled toxicants, thus representing a new and fascinating application in risk assessment strategies. PMID- 16353344 TI - Paraoxonase (PON1): from toxicology to cardiovascular medicine. AB - Paraoxonase (PON1) is a liver and plasma enzyme most studied because of its ability to hydrolyze the active metabolites of several organophosphorus insecticides. The discovery that PON1 can also metabolize oxidized phospholipids has spurred research on its possible role in coronary heart disease and atherosclerosis. Additionally, its potential roles in metabolizing pharmaceutical drugs and microbial quorum sensing factors are also being explored. PON1 displays several polymorphisms that influence both its level of expression and its catalytic activity, thus determining the rates at which a given individual will detoxify a specific insecticide, metabolize harmful oxidized lipids, and activate or inactivate specific drugs and quorum sensing factors. PMID- 16353345 TI - Tissue-specific lipoprotein lipase: relationships to body composition and body fat distribution in normal weight humans. AB - Twenty-six normal weight subjects (22 female, 4 male) were studied to determine the relationships of fasting levels of lipoprotein lipase in gluteal adipose tissue (ATLPL) and skeletal muscle (SMLPL) to body composition and body fat distribution. No relationship was found between fasting gluteal ATLPL and percent (%) body fat. There was, however, an inverse relationship between fasting SMLPL (from the vastus lateralis) and %body fat (p=0.005). A strong inverse correlation was also seen between fasting ATLPL and waist/hip ratio (p=0.0006), a measurement of body fat distribution. These relationships existed with or without the male subjects included. The tissue-specific relationships of lipoprotein lipase to body composition and body fat distribution could relate to the development of obesity or the maintenance of normal body weight by the effects of the lipase on the partitioning of lipoprotein triglyceride fatty acids. PMID- 16353346 TI - Insulin levels, physical activity, and urinary catecholamine excretion of obese and non-obese rhesus monkeys. AB - The hypothesis that spontaneous obesity in rhesus monkeys is associated with abnormalities in energy expenditure was tested. Obese (n=7) and non-obese (n=5) monkeys were described in terms of body size and composition, food intake, and physical activity. Additionally, the relationships among fasting and stimulated insulin levels in serum, C-peptide levels in serum and urine, and urinary catecholamines were examined. Obese animals had primarily abdominal deposition of excess body fat, as indicated by markedly elevated abdominal circumferences and skin-fold thicknesses. Food intake did not differ between groups. Physical activity was much lower in the obese group. Obese monkeys had markedly higher serum insulin and C-peptide levels in the fasted state and in response to an intravenous glucose challenge. Urinary excretion of C-peptide and catecholamines was measured during successive 2-day periods of ad libitum feeding, food deprivation, and refeeding in order to examine potential differences between groups in sympathoadrenal activity and their relationship to insulin secretion. C peptide excretion was greater for obese and decreased for both groups during food deprivation. Urinary dopamine (DA), norepinephrine (NE), and epinephrine (E) levels were significantly greater for obese animals in all conditions. DA excretion was lowest during deprivation and E excretion was lowest during refeeding, whereas NE excretion was relatively unaffected by feeding condition. The overall patterns of C-peptide and catecholamine excretion were qualitatively similar for both groups, and there were no reliable differences between obese and non-obese in their responses to the feeding manipulation. The results suggest that hyperinsulinemia associated with obesity in rhesus monkeys is linked to increased catecholamine secretion and a resistance to catecholaminergic action. PMID- 16353348 TI - Meal pattern analysis of macronutrient intake after PVN norepinephrine and peripheral clonidine administration. AB - Norepinephrine (NE) injected into the paraventricular nucleus (PVN) of the hypothalamus of rats is a potent stimulant of food intake, more specifically ingestion of the carbohydrate nutrient. In 2 experiments of the present study, this effect was found to be dose-dependent, and the effectiveness of NE in potentiating total food consumption was greatly reduced when the carbohydrate diet was removed. In addition, experiments using a computer-automated data acquisition apparatus were performed to characterize, in detail, the impact of PVN injection of NE and peripheral administration of the alpha2-noradrenergic agonist clonidine (CLON) on the macrostructure of feeding behavior in animals given 3 pure macronutrient diets. These 2 compounds, injected at the onset of the nocturnal feeding cycle, had very similar effects on meal patterns, with both affecting nutrient intake by increasing meal size and duration rather than by increasing meal frequency. They both affected primarily the first meal of the dark cycle, selectively enhancing carbohydrate ingestion by increasing Kcal intake, percent composition in the total diet and feeding time, and also by decreasing the satiating impact of this macronutrient. These stimulatory effects of NE and CLON on carbohydrate ingestion during the first meal were followed by complete recovery over the next 1 to 2 hours after injection. In addition to these predominant effects on carbohydrate intake, PVN NE at the highest doses tested (10 and 20 nmoles) produced a small increase in fat intake, whereas peripheral CLON actually decreased intake of fat and protein over the 12-hour cycle. The similarities in the impact of NE and CLON on carbohydrate feeding patterns support the hypothesis that both agonists may be acting via the same PVN alpha2-noradrenergic system controlling ingestion of the carbohydrate-rich meals which predominate at dark onset. PMID- 16353349 TI - Time-dependent effects of progressive gamma-linolenate feeding on hyperphagia, weight gain, and erythrocyte fatty acid composition during growth of Zucker obese rats. AB - Obese Zucker rats (fa/fa) have low levels of arachidonic acid (AA) in liver phospholipids (PL). We have previously shown that a 70% gamma-linolenate concentrate (GLA; an AA intermediate) fed at a fixed dose (0.07 g/day) normalized hepatic PL AA and reduced weight gain selectively in the obese animals. In a follow-up study, 16 obese (fa/fa) and 16 lean (Fa/Fa) 4-week-old male rats were randomized into 4 groups of 8 each and gavaged daily with soybean oil (SOY) containing 55% 18:2omega6 (an AA precursor) or GLA, using a progressive dose (< or = 5% of total calories) based on body weight. A defined diet with 11% of energy as SOY was fed ad libitum for 60 days. GLA obese had lower body weight (p<0.0001) and 60-day cumulative food intake (p<0.05) compared to SOY obese, but neither parameter differed between the lean groups. For the last twenty days cumulative food intake was identical for GLA obese and SOY lean, whereas SOY obese consumed 18% more (p<0.05). Thus the progressive dose of GLA selectively suppressed hyperphagia in obese Zucker rats. Erythrocytes collected at 15-day intervals showed parallel increases in AA in both genotypes over time, suggesting normal AA availability during rapid growth. Thus, the reduced PL AA in the livers from the obese rats probably reflects impaired distribution in selected tissues rather than reduced hepatic production. Due to the potential health risks of enriching tissue lipids with AA, great caution is advised in considering GLA as therapy for human obesity. PMID- 16353347 TI - Mechanism of free fatty acid effects on hepatocyte insulin receptor binding and processing. AB - We determined whether the palmitate effects on hepatocyte insulin receptor binding and post-receptor trafficking were mediated by accelerated mitochondrial beta-oxidation or accumulation of intracellular fatty acyl-CoA derivatives and possibly protein acylation. Preincubation of hepatocytes with moderate concentrations of palmitate (0.5 mM) resulted in a 23% decline in cell-surface binding and proportional decreases in receptor-mediated insulin internalization and degradation. Brief pretreatment of hepatocytes with the carnitine palmityltransferase-I inhibitor, methyl palmoxirate (MP), prevented 70% of the palmitate effects. At higher palmitate concentrations (2.0 mM), cell-surface binding was reduced by 34%, whereas internalization of the receptor complex was reduced by 78%. These effects were only partially prevented by MP pretreatment. Receptor-mediated insulin degradation increased by 34% and was uninfluenced by MP pretreatment. Octanoate, which is rapidly shunted into mitochondrial oxidation, produced a dose-dependent reduction in insulin binding, with proportional decreases in internalization and degradation. Similarly preincubation with 2.0 mM oleate, which, unlike palmitate, is not known to produce protein acylation, resulted in proportional decreases in insulin receptor binding and receptor mediated internalization and degradation. High concentrations of octanoate or oleate (2.0 mM) did not reproduce the additive post-receptor effects of palmitate. We conclude that the receptor and post-receptor effects of moderate palmitate concentrations are closely linked to accelerated fatty acid oxidation. The post-receptor effects observed at higher concentrations involve other mechanisms, possibly relating to intracellular levels of palmityl-CoA derivatives. PMID- 16353350 TI - Obesity in families of extremely obese women. AB - In order to assess the prevalence of obesity in families of extremely obese individuals, we conducted a mail survey of a national obesity organization. Thirty-nine percent (N=981) of the questionnaires were completed and returned. Respondents were excluded from further analysis if they were adopted, male, their gender could not be determined, provided incomplete information about their parents or their own height and weight, or were less than 22 or greater than 63 years of age. The analyses included 729 probands and their families. Both the prevalence and the extent of obesity were high in the families members. The average family members' body mass index (BMI=kg/m2) was 30, and 78% of the families studied had at least one other obese (BMI>30 kg/m2) first-degree relative (parent, sibling, or child). Although obesity was common in the families, survey respondents were generally the heaviest members of their families, having an average BMI of 47 kg/m2. Correlations among first-degree relatives were similar to those found for average weight groups, suggesting that obesity and BMI are similarly influenced by family genetic factors in this extremely obese population. PMID- 16353351 TI - Feeding response of rats to no-fat and high-fat cakes. AB - The nutritional effects of high-fat diets have been extensively studied in laboratory animals, but as yet few experiments have examined the feeding response of animals to newly developed fat substitutes. The present study used commercially available no-fat (0% fat, 92% carbohydrate) and high-fat (41% fat, 54% carbohydrate) cake to determine the effects of fat substitutes on food preference and caloric intake in rats. The first experiment showed that nondeprived rats found the high-fat and no-fat cakes equally palatable and highly preferred to lab chow. Food deprived rats, however, preferred the high-fat cake to the no-fat cake, which may be related to its higher caloric density. In the second experiment, rats fed high-fat cake, in addition to chow, for 30 days consumed more calories and gained more weight than did rats fed no-fat cake and chow. The no-fat cake group, however, overate and gained more weight than chow only controls. The hyperphagic response to the no-fat cake can be attributed to its carbohydrate content, moisture, and high palatability. Thus, removing fat from the cake reduced, but did not eliminate, its obesity-promoting effect. Obviously, low-fat foods must be consumed in moderation if used for weight control. PMID- 16353353 TI - Effect of enterostatin on the feeding responses to galanin and NPY. AB - We have investigated the possibility that enterostatin may inhibit the intake of dietary fat by inhibiting either galanin or NPY-induced feeding pathways. Rats, adapted to either high fat (HF) or low fat-high carbohydrate (HC) diets and fitted with third ventricular cannulas were used to study the effects of intracerebroventricular (i.c.v.) enterostatin on i.c.v. NPY and galanin induced feeding responses in satiated rats. An equimolar dose of enterostatin (0.1 nmoles) inhibited, while a tenfold excess of enterostatin abolished the feeding response to galanin in rats adapted to a HF diet. The galanin stimulation of food intake was reduced in rats adapted to the HC diet and this response was less sensitive to inhibition by enterostatin. Enterostatin had no inhibitory effects on NPY-induced feeding in rats adapted to the HC diet and only a small inhibitory effect, at tenfold molar excess, in rats adapted to the HF diet. The ability of enterostatin to bind to galanin or NPY Y-1 receptors was investigated in ligand binding studies. Enterostatin failed to displace 125I-galanin or 125I-PYY from specific binding sites in rat forebrain homogenates or SK-N-MC cells respectively. The data provide support for the hypothesis that enterostatin specifically inhibits a galanin-responsive fat intake system, but indicate that this effect is not modulated by direct interaction with either galanin or NPY-Y1 receptors. PMID- 16353352 TI - Non linear weight gain with long term overfeeding in man. AB - This study deals with the pattern of body weight gain during an overfeeding period with a constant energy intake, in order to assess whether total daily energy expenditure (TEE) increased with body weight and thus could account for the progressive slow down in body weight gain over time. Twenty-four young adult males (12 pairs of identical twins) were overfed by 4.2 MJ per day, six days a week, for a total of 84 days during a 100-day overfeeding period. The total excess amount each man consumed was 353 MJ. It was assumed that, at a given time, the TEE increase (E) was dependent on body weight gain and energy cost (C) was proportional to the daily body weight gain. Results show an exponential increase in body weight, fat free mass, and fat mass (with half-times of 86, 57, and 84 days, respectively) that allows the calculation of E (246 +/- 37 kJ x kg(-1) x d( 1), mean +/- SE) and C (32.3 +/- 2.4 MJ x kg(-1)). Energy expenditure from other sources besides resting metabolic rate, such as physical activity and thermic effect of food, may represent as much as 65% of E. At the beginning of the overfeeding period, almost all the energy surplus was recovered as body substances but this proportion decreased to 60% after 100 days of overfeeding. It is concluded that 1) TEE changes were related to body weight change, 2) about 65% of E were accounted for by physical activity, thermic effect of food, or some other components, and 3) the fraction of the energy surplus stored as body substances decreased with the duration of overfeeding. PMID- 16353354 TI - Effect of ethnicity and geographical location on body weight, dietary restraint, and abnormal eating attitudes. AB - Previous studies have examined the effect of ethnicity on obesity, concerns about shape and weight, and attitudes about eating. We hypothesized that geographical location would also influence these variables, and that students growing up in the northern part of the United States and attending northern colleges would differ from students from the South. To examine this, we studied a random sample of 275 African-Americans (AA) and 224 white college students in the entering class of two northern colleges (University of Pittsburgh or University of Massachusetts) or two southern colleges (Augusta or Paine College). All subjects were weighed and completed the Revised Restraint Scale and the EAT-26. AA women were heavier than white women, with no differences due to geographical location. Despite being thinner, white women reported more dietary restraint than AA women. This difference between AA and white women was apparent in both northern and southern college students. In contrast, geographical location was the strongest determinant of bulimic attitudes; both men and women at northern colleges reported higher bulimia scores than those at southern schools. Thus ethnicity appears to be a major determinant of body weight and attitudes about shape and dieting, whereas geographical location appears to exert greater influence on bulimic attitudes. PMID- 16353355 TI - Decreased hepatic insulin extraction precedes overt noninsulin dependent (Type II) diabetes in obese monkeys. AB - Many obese middle-aged rhesus monkeys (Macaca mulatta) spontaneously develop noninsulin dependent diabetes mellitus (NIDDM). Basal hyperinsulinemia and increased stimulated plasma insulin levels are associated with this obesity and precede the onset of overt diabetes. The present studies sought to determine the relative contributions of enhanced insulin secretion and of reduced insulin clearance to this early obesity-associated hyperinsulinemia. Direct simultaneous measurement of portal and jugular vein insulin levels in two normal monkeys showed a constant rate of hepatic insulin extraction of 56+/-3% over the range of peripheral insulin levels from 351+/-113 to 625+/-118 pmol/L. In 33 additional monkeys ranging from normal to diabetic, basal C-peptide levels were examined as an indicator of beta-cell secretion and the molar ratio of plasma C-peptide to insulin (C/I ratio) under basal steady state conditions calculated as an index of hepatic insulin extraction. Well in advance of overt diabetes, there was a progressive decline of 67% in the apparent hepatic insulin extraction rate in association with increased obesity and plasma insulin levels. Basal insulin levels and hepatic insulin extraction returned toward normal in monkeys with impaired glucose tolerance and in those with overt diabetes. We conclude that reduced insulin disposal, probably due to reduced hepatic extraction of insulin, in addition to increased beta-cell activity, contributes to the development of basal hyperinsulinemia in obese rhesus monkeys progressing toward NIDDM. In addition, in overt diabetes, normal hepatic insulin extraction in the presence of limited beta-cell secretion may exacerbate the hypoinsulinemic state. PMID- 16353356 TI - Familial aggregation of morbid obesity. AB - Recent studies have shown major gene effects for obesity in randomly ascertained families. To investigate the familial aggregation of a specific subset of obesity, which is particularly prone to medical complications, families with morbid obesity were studied. This condition occurs in 1%-2% of the population and is defined as 45.5 kg (100 pounds) or more over ideal weight. First-degree relatives of 221 morbidly obese probands (1560 adults) were identified, and height and weight (current and greatest) were obtained from each family member. Morbid obesity occurred in the family members of the probands 8 times more often than in the general population. Of the morbidly obese probands, 48% had one or more first-degree relatives who were also morbidly obese compared to a 6% population estimate. By the ages of 20-24, 12% of the morbidly obese probands were already 45.5 kg or more overweight, and 45% were 22.7 kg (50 pounds) or more overweight. There was little difference in the prevalence of familial morbid obesity by the gender of the probands: 47% of the male probands and 48% of the female probands had another morbidly obese relative, while 67% and 53% of the early onset (before age 25) male and female probands, respectively, had one or more first-degree relatives who were also morbidly obese. In addition to the extreme degree of familial aggregation, the prevalence of morbid obesity in parent-offspring sets was calculated within the morbidly obese families. Morbidly obese families who have one or two morbidly obese parents have a 2.6 times increased risk (p<0.002) of having one or more morbidly obese adult offspring, compared to families who have neither parent morbidly obese. Evidence for trimodality of the body mass index distribution was found for each gender (p = 0.0006 for male relatives and p = 0.075 for female relatives). The strong familial aggregation of morbid obesity indicates the need for further understanding of the genetic determinants of this extreme clinical disorder and how environmental factors affect the genetic expression of the trait. PMID- 16353357 TI - BSB: a new mouse model of multigenic obesity. AB - We report here a new mouse model of multigenic obesity. Backcross progeny ((C57BL/6J x Mus spretus)F1 x C57BL/6J), designated as BSB mice, range from 1% to 50% body fat. Since both parental strains are relatively lean, the wide range of the phenotype in the BSB mice indicates the involvement of multiple genes to produce obesity. Obesity in BSB mice results from increases in both intra abdominal and subcutaneous fat and is associated with hyperinsulinemia, hyperglycemia, and hyperlipidemia. Female and male BSB mice do not differ in the degree of obesity obtained. Stimulated plasma corticosterone levels are reduced in obese male and female mice. The development of appropriate genetic markers and statistical methods have made it feasible to analyze quantitative polygenic traits in animal models by employing F2 or backcross progeny. Thus, this BSB model is uniquely suited to the genetic analysis of the multifactorial quantitative trait of obesity and its associated phenotypes. PMID- 16353358 TI - Sympathetic activity, age, sucrose preference, and diet-induced obesity. AB - Only half the adult male Sprague-Dawley rats which are placed on a diet relatively high in calories, fat, and sucrose (HE diet) develop diet-induced obesity (DIO). The rest are diet-resistant (DR). Some chow-fed rats prone to develop DIO on an HE diet have greater initial food intake of this diet and all have greater glucose-induced plasma norepinephrine (NE) increases than DR-prone rats. Here we looked for a relationship of sucrose preference or 24-hour urinary catecholamine excretion as possible phenotypic markers of the DIO- and DR-prone states before HE diet exposure as a function of age. When begun on an HE diet at 3 months of age, DIO-prone rats gained 30% more weight over 3 months than DR prone rats and had 35% heavier retroperitoneal fat pads. While still on chow, sucrose preferences were similar, but 24 hour urine NE levels were 29% higher in DIO- than in DR-prone rats. The slope of the curve of urine NE versus body weight gain after 3 months on HE diet was 4-fold greater in DIO- than in DR-prone rats. After 3 months on the HE diet, there was no statistical relationship between 24 hour urine NE and body weight or prior body weight gain in DIO or DR rats. Six month-old DIO-prone rats had 126% and 128% more urine NE and gained 112% and 232% more weight after 3 months on HE diet than DR-prone and chow-fed rats, respectively. Only DIO-prone rats showed a correlation (r=0.879; p=0.05) between urine NE levels and subsequent weight gain on HE diet. Thus, 3- or 6-month-old DIO- and DR-prone rats can be identified by their 24-hour basal urine NE levels but not sucrose preference prior to HE diet exposure. While this may suggest higher basal sympathetic activity in DIO-prone rats, other explanations are possible. PMID- 16353359 TI - Segregation analysis of body mass index in an unselected French-Canadian sample: the Quebec Family Study. AB - Interest in a single gene etiology for obesity, as assessed by the body mass index (BMI), has been spurred recently by reports of a putative recessive major gene for extreme values, which accounts for as much as 40% of the variance. The major gene hypothesis was evaluated here in the Quebec Family Study, a random sample of 375 French-Canadian volunteer families. This report represents one component in a more complete investigation of obesity in these families. In contrast to the recent studies, a major gene hypothesis for BMI was not verified here. Although there was a major effect, it did not conform to a Mendelian pattern of transmission. A multifactorial component (i.e., polygenic and/or common environmental factors) accounted for 42% of the phenotypic variance. In addition, evidence of heterogeneity between the generations was found. The heterogeneity was traced to the major non-Mendelian component (which accounted for 0.01% of the variance in parents and over 40% in offspring) rather than to the multifactorial one. These results would suggest that a simple recessive gene mixed model may not be sufficient to explain the familial distribution of the BMI. Several factors which may have contributed to these results include temporal trends and surrogate effects such as those related to variation in body composition and energy balance components. PMID- 16353360 TI - Evidence of commingling in human eating behavior. AB - This investigation tested whether distributions of certain aspects of eating behavior were consistent with the notion of a "mixture model;" that is, two or more distinct commingled component distributions, consistent with the possibility of major gene action. Undergraduates (n=901) completed self-report trait measures of hunger, disinhibition, and dietary restraint. Variables were residualized for gender and age and transformed to remove skewness. Residualized transformed distributions were tested for departure from unimodality with Hartigan's dip statistic. The distributions of all three aspects of eating behavior were significantly non-unimodal. Next, component multivariate normal distributions were estimated via maximum likelihood. Likelihood ratio tests were employed to compare nested models. A mixture of four distributions with unequal variance covariance matrices fit significantly better than any more parsimonious model. In sum, these data strongly suggest that the distributions of several measures of eating behavior are composed of four component distributions. This finding is consistent with the possibility of major gene effects for eating behavior. PMID- 16353361 TI - Diversity of insulin resistance in monkeys with normal glucose tolerance. AB - Insulin resistance has been proposed as a critical factor in the development of Type II diabetes, hypertension, dyslipidemia, and coronary artery disease. However, even in normal healthy individuals, a wide range of in vivo insulin action has been found. In the present study we sought to examine this heterogeneity in insulin action in both normal and spontaneously obese nonhuman primates. Maximal insulin responsiveness as measured by a hyperinsulinemic euglycemic clamp, fasting plasma glucose, and insulin levels, beta-cell insulin response to glucose, glucose tolerance, and adiposity were measured in 22 male rhesus monkeys. Results showed that lean animals (body fat < or = 22%) had higher insulin-stimulated glucose uptake (M rate: 14.42+/-1.8 mg/kg FFM/min) compared to obese (8.08+/-0.8). The obese monkeys, with 23-49% body fat, had a wide range of M values (5.32-14.29 mg/kg FFM/min) which showed no relationship to degree of adiposity. In all monkeys, M values had a strong inverse correlation with fasting plasma insulin levels (r=-0.76; p<0.001), but not with fasting glucose or glucose disappearance rate. We conclude that neither degree of obesity above a critical threshold nor range of glucose tolerance is related to insulin resistance; however, in individuals with normal glucose tolerance an early reliable indicator of defective insulin action appears to be fasting insulin concentration. Longitudinal determination of basal insulin levels obtained under standardized conditions so as to minimize extraneous variability is likely to strengthen the ability to predict insulin resistance and possible later development of overt Type II diabetes. PMID- 16353362 TI - Tonic sympathetic nervous system inhibition of insulin secretion is diminished in obese Zucker rats. AB - It has long been known that the central nervous system (CNS) directly affects pancreatic insulin release. This study was undertaken to determine the effect of the CNS on pancreatic insulin release in three-month-old female lean (Fa/Fa) and hyperinsulinemic obese (fa/fa) Zucker rats. Chloral hydrate (400 mg/kg) was used as the anesthetic agent. The in situ brain-pancreas perfusion model with intact pancreatic innervation was used in this investigation. The study measured insulin secretion in response to a 60-minute glucose stimulus (200 mg/dl). CNS-intact and CNS-functionally ablated obese and lean rats were used. During the 60-minute perfusion period significantly more insulin was released by pancreata from obese rats compared to those from lean rats. In lean rats, about twice as much insulin was released by pancreata from CNS-ablated rats than from CNS-intact rats (P < 0.05), demonstrating a CNS tonic inhibition of insulin secretion. In obese rats, there was no significant difference in insulin released by the pancreata of the CNS-intact and CNS-ablated rats. To determine if there was a masking effect of predominant PNS activity over the SNS in the CNS-intact obese rats, bilateral vagotomy was performed in a group of otherwise CNS-intact obese rats prior to the onset of perfusion. Tonic inhibition was still not observed in the CNS vagotomized obese rats. In conclusion, hypersecretion of insulin in obese rats is partially due to diminished tonic sympathetic nervous system inhibition of insulin release. These results provide additional evidence regarding abnormal CNS control of insulin secretion in obese Zucker rats. PMID- 16353363 TI - Adrenalectomy and castration in the genetically obese (ob/ob) mouse. AB - The present studies have tested the hypothesis that adrenalectomy could modify the phenotypic expression of genetic obesity by examining the effects of adrenalectomy on the function of the gonadal system in lean and ob/ob mice. Corticosterone concentrations were undetectable in the adrenalectomized animals. Adrenalectomy significantly slowed the weight gain of obese mice in comparison to sham-adrenalectomized controls. Gonadectomy had no independent effect on weight gain. The testes, prostate, and seminal vesicles in the ob/ob mice were significantly smaller than in the lean animals. Castration lowered the weights of the prostate and seminal vesicles in the lean mice to weights close to those observed in the castrated ob/ob mice. Castration significantly increased the concentrations of LH and FSH in both ob/ob and lean mice, but the absolute concentrations were higher in the lean mice in both conditions. Adrenalectomy per se had no effect on the concentration of LH, FSH, or testosterone or on the weights of the prostate or seminal vesicles. These data indicate that adrenalectomy has no effect on the physiologic control of the reproductive system in genetically obese mice, and are consistent with the hypothesis that the defect in the ob/ob mouse is a modulator of steroid action which over expresses glucocorticoid effects and under expresses gonadal steroid effects. PMID- 16353364 TI - Weight cycling: a review of the animal literature. AB - This review critically evaluates the animal literature concerning the effects of weight cycling on factors related to development of obesity, diabetes, hypertension, and hyperlipidemia. Although human studies have been used to retrospectively examine the relationship between fluctuations in body weight and a variety of disease markers, direct causal links between weight cycling and negative health effects have been inferred from a series of scientific publications using animals as subjects. We use data from 24 such publications to evaluate evidence for and against a series of hypotheses that have been suggested regarding weight cycling and health. Although there are some intriguing results, there is currently little evidence to support any of these hypotheses. However, methodological limitations were identified in many of these studies, and caution should be used in making definitive decisions about weight cycling. Weight cycling studies could be improved by including more appropriate controls, comparing controls to weight cycling animals at more appropriate time points, and giving more attention to potential effects of diet composition. While more careful research is needed, at this time we conclude that the published animal literature does not justify any warnings about the hazards of weight cycling. PMID- 16353365 TI - Visual and tactile guidance of dexterous manipulation tasks: an fMRI study. AB - Models of motor guidance that dynamically adjust to the availability and quality of sensory information are based on the observation that dexterous tasks are routinely performed using various combinations of visual and tactile inputs. However, a dynamic neural system that acquires and processes relevant visual and tactile information remains relatively uncharacterized in humans. In this study, whole-brain functional magnetic resonance images were acquired during a dexterous manipulation task, compression of the end caps of a slender spring prone to buckling, to investigate the neural systems associated with motor guidance under four visual and tactile guidance conditions: (1) eyes closed (no visual input), smooth end caps, (2) eyes dosed, rough end caps, (3) eyes open and watching hand, smooth end caps, and (4) eyes open and watching hand, rough end caps. Performance of the dexterous task remained constant in all conditions. Variations in the two levels of visual input resulted in modulation of activity in the middle and inferior occipital gyrii and inferior parietal lobule, and variation in the two levels of tactile input during the task resulted in modulation of activity in the precentral (primary motor) gyrus. Although significantly active in all conditions, cingulate gyrus, medial frontal gyrus, postcentral gyrus, and cerebellum activities were not modulated by levels of either visual or somatosensory input, and no interaction effects were observed. Together, these data indicate that a fine-tuned motor task guided by varying visual and tactile information engages a distributed and integrated neural complex consisting of control and executive functions and regions that process dynamic sensory information related to guidance functions. PMID- 16353366 TI - Sentence processing in frontotemporal dementia. AB - Patients with frontotemporal dementia (FTD) have sentence comprehension difficulty. We examined the hypothesis that both grammatical and resource factors contribute to their impaired sentence comprehension with a traditional, off-line sentence comprehension task, and an on-line sentence processing procedure that minimizes task-related resources. This was investigated in subgroups of FTD patients with Progressive Non-fluent Aphasia (PNFA; n = 5) who have effortful speech; non-aphasic patients with an executive deficit (EXEC; n = 8); and Semantic Dementia (SD; n = 3) patients with poor single word comprehension. The results were correlated with measures of executive resources. We found that PNFA patients are significantly impaired in their off-line sentence comprehension, and that their performance correlated with auditory-verbal short-term memory. PNFA patients also demonstrated a pattern of slowed processing for the on-line sentence measure. This is consistent with the hypothesis that information relevant for constructing sentence representations during comprehension degrades in working memory as it is activated over an abnormally slowed time course. EXEC patients had modest off-line sentence comprehension difficulty, and this correlated with performance on measures of working memory, planning, and inhibitory control. On-line processing in EXEC patients demonstrated their insensitivity to sentence-based information. This raises the possibility that a limitation in material-neutral executive resources not dedicated to grammatical processing may play a role in their sentence comprehension deficit. SD patients' pattern of on-line sentence comprehension paralleled control subjects' performance. We conclude that grammatical and executive components both contribute to sentence comprehension, and that the profile of sentence comprehension difficulty varies across FTD subgroups depending on the sentence processing component that is impaired. PMID- 16353367 TI - Bilateral thalamic lesions affect recollection- and familiarity-based recognition memory judgments. AB - The contribution of the thalamus to different forms of explicit memory is poorly understood. In the current study, explicit memory performance was examined in a 40-year-old male (RG) with bilateral anterior and medial thalamic lesions. Standardized tests indicated that the patient exhibited more severe recall than recognition deficits and his performance was generally worse for verbal compared to nonverbal memory. Recognition memory tests using the remember-know (R/K) procedure and the confidence-based receiver operating characteristic (ROC) procedure were used to examine recollection- and familiarity-based recognition. These tests revealed that RG had deficits in recollection and smaller, but consistent deficits in familiarity. The results are in agreement with models indicating that the anteromedial thalamus is important for both recollection- and familiarity-based recognition memory. PMID- 16353368 TI - Multiregional cell assemblies, temporal binding and the representation of conceptual knowledge in cortex: a modern theory by a "classical" neurologist, Carl Wernicke. AB - A contemporary view of conceptual representation in the brain holds that conceptual knowledge is distributed throughout the cerebral cortex, localized to cortical regions involved in their initial processing, and functionally interconnected through synchronized associative processes that are mediated through "convergence zones". The primary goal of the present paper is to point out that Carl Wernicke proposed a theory of how concepts are acquired and represented in cortex which is strikingly similar to contemporary views. Wernicke sketched his ideas on this topic in his earliest writings on aphasia. But his theory is developed most fully in the Grundriss der Psychiatrie (Outlines of Psychiatry), published in 1900 and never translated into English. We describe Wernicke's views on the distributed nature of conceptual knowledge in the brain using select quotes from his early work, and by providing a translation of relevant sections of the Grundriss der Psychiatrie. PMID- 16353369 TI - Alcohol use and alcohol-related problems among lesbians and gay men. AB - While a substantial amount is known about some of the risk factors for alcohol related problems among lesbians and gay men, major gaps in knowledge exist. Epidemiological studies focusing on alcohol use rarely ask about sexual orientation, and broad-based studies of sexual minority population groups have only occasionally assessed alcohol use. Although the AIDS crisis has stimulated substantial research on alcohol and other substance use among gay men, only a handful of studies have systematically explored lesbians' use of alcohol. Further, existing research on sexual orientation and alcohol use is characterized by a plethora of methodological problems. Nevertheless, when viewed as a whole, this research suggests that lesbians and gay men are more likely than their heterosexual counterparts to drink alcohol and to report alcohol-related problems; differences based on sexual orientation are more pronounced for women than for men. Risks related to alcohol use do not stem from sexual orientation per se, but are more likely a consequence of cultural and environmental factors associated with being part of a stigmatized and marginalized population. Much of the research on alcohol use among sexual minorities has focused on White, middle class, and well-educated lesbians and gay men. There is a clear need for more research with bisexual women and men and with sexual minority members of color. Longitudinal studies, including those that focus on treatment effectiveness, are particularly lacking. PMID- 16353370 TI - Intrathoracic airway trees: segmentation and airway morphology analysis from low dose CT scans. AB - The segmentation of the human airway tree from volumetric computed tomography (CT) images builds an important step for many clinical applications and for physiological studies. Previously proposed algorithms suffer from one or several problems: leaking into the surrounding lung parenchyma, the need for the user to manually adjust parameters, excessive runtime. Low-dose CT scans are increasingly utilized in lung screening studies, but segmenting them with traditional airway segmentation algorithms often yields less than satisfying results. In this paper, a new airway segmentation method based on fuzzy connectivity is presented. Small adaptive regions of interest are used that follow the airway branches as they are segmented. This has several advantages. It makes it possible to detect leaks early and avoid them, the segmentation algorithm can automatically adapt to changing image parameters, and the computing time is kept within moderate values. The new method is robust in the sense that it works on various types of scans (low-dose and regular dose, normal subjects and diseased subjects) without the need for the user to manually adjust any parameters. Comparison with a commonly used region-grow segmentation algorithm shows that the newly proposed method retrieves a significantly higher count of airway branches. A method that conducts accurate cross-sectional airway measurements on airways is presented as an additional processing step. Measurements are conducted in the original gray-level volume. Validation on a phantom shows that subvoxel accuracy is achieved for all airway sizes and airway orientations. PMID- 16353371 TI - Matching and anatomical labeling of human airway tree. AB - Matching of corresponding branchpoints between two human airway trees, as well as assigning anatomical names to the segments and branchpoints of the human airway tree, are of significant interest for clinical applications and physiological studies. In the past, these tasks were often performed manually due to the lack of automated algorithms that can tolerate false branches and anatomical variability typical for in vivo trees. In this paper, we present algorithms that perform both matching of branchpoints and anatomical labeling of in vivo trees without any human intervention and within a short computing time. No hand-pruning of false branches is required. The results from the automated methods show a high degree of accuracy when validated against reference data provided by human experts. 92.9% of the verifiable branchpoint matches found by the computer agree with experts' results. For anatomical labeling, 97.1% of the automatically assigned segment labels were found to be correct. PMID- 16353372 TI - A novel local thresholding algorithm for trabecular bone volume fraction mapping in the limited spatial resolution regime of in vivo MRI. AB - Recent advances in micro-magnetic resonance imaging have shown the possibility of in vivo assessment of trabecular bone architecture. However, the small feature size and relatively low signal-to-noise ratio (SNR) achievable in vivo cause the intensity histogram to be unimodal. The critical first step in the processing of these images is the extraction of bone volume fraction for each voxel. Here, we propose a local threshold algorithm (LTA) that determines the marrow intensity value in the neighborhood of each voxel based on nearest-neighbor statistics. Using the local marrow intensities we threshold the image and scale the intensities of voxels partially occupied by bone to produce a marrow volume fraction map of the trabecular bone region. We show that structural parameters derived with the LTA are highly correlated with those obtained with the previously published histogram deconvolution algorithm (HDA) and that the LTA is robust to image noise corruption. The LTA is found to correctly identify trabeculae with a significantly higher reliability than HDA. Finally, we demonstrate that the LTA is superior in preserving connectivity by showing for 75 in vivo images that the genus of the trabecular bone surface is always higher than when processed with the HDA. PMID- 16353373 TI - Decision strategies that maximize the area under the LROC curve. AB - For the 2-class detection problem (signal absent/present), the likelihood ratio is an ideal observer in that it minimizes Bayes risk for arbitrary costs and it maximizes the area under the receiver operating characteristic (ROC) curve [AUC]. The AUC-optimizing property makes it a valuable tool in imaging system optimization. If one considered a different task, namely, joint detection and localization of the signal, then it would be similarly valuable to have a decision strategy that optimized a relevant scalar figure of merit. We are interested in quantifying performance on decision tasks involving location uncertainty using the localization ROC (LROC) methodology. Therefore, we derive decision strategies that maximize the area under the LROC curve, A(LROC). We show that these decision strategies minimize Bayes risk under certain reasonable cost constraints. The detection-localization task is modeled as a decision problem in three increasingly realistic ways. In the first two models, we treat location as a discrete parameter having finitely many values resulting in an (L + 1) class classification problem. In our first simple model, we do not include search tolerance effects and in the second, more general, model, we do. In the third and most general model, we treat location as a continuous parameter and also include search tolerance effects. In all cases, the essential proof that the observer maximizes A(LROC) is obtained with a modified version of the Neyman-Pearson lemma. A separate form of proof is used to show that in all three cases, the decision strategy minimizes the Bayes risk under certain reasonable cost constraints. PMID- 16353374 TI - Resolution-effective diameters for asymmetric-knife-edge pinhole collimators. AB - The effects of penetration are included in the formulas for the prediction of the resolution of pinhole collimators through the use of effective diameters. Expressions of the resolution-effective diameter for pinholes with a double-knife edge (DKE) profile are available in the literature. In this paper the expressions applicable to asymmetric-knife-edge (AKE) profiles, which include the important case of the single-knife-edge (SKE), are presented. Results indicate that the simplest methods that are still accurate in the calculation of DKE effective diameters do not produce in general formulas with similar accuracy for AKE profiles, due to increased susceptibility to penetration. Especially at high energy (365 keV), for the SKE case more advanced formulas are necessary and were, therefore, derived. PMID- 16353375 TI - Managing financial conflicts of interest in research. AB - Evidence points to commercial sponsorship and personal financial gains for researchers leading to reporting more favorable results, tainting the climate of academic integrity, and negative public perceptions. Research institutions attempt to protect their own reputations and those of their faculty through establishing thresholds for reporting financial involvement and through committee review that may suggest that the sponsoring organization impose management practices designed to reduce conflicts or declining the funding. In one prominent university research system, a quarter of research projects reviewed required management because of conflicts of interest, the most common of these stemming from high "consulting fees" paid to researchers. The degree of understanding of policies regarding conflicts among researchers is uneven, and some regard these as private matters. Differences of opinion exist in the research community over whether disclosure of financial interests, although necessary, is sufficient to ensure a reasonable level of freedom from bias and to maintain public trust. PMID- 16353376 TI - HPV testing and cervical cancer screening: recommendations and practice patterns. AB - In summary, guidelines for cervical cancer screening continue to evolve, as knowledge of the pathogenesis of the disease as well as the role of HPV expands. Oncogenic HPV typing has proven effective in cancer screening and follow-up in certain situations, and its uses will undoubtedly increase. Patients and providers may be reassured with negative HPV testing, but long term management of positive HPV testing (especially in conjunction with negative cytology) is unclear. At present, however, numerous worldwide, prospective studies involving HPV testing and cervical cancer screening will, we hope, provide some of the answers regarding optimal management of women who remain persistently HPV positive. PMID- 16353377 TI - Adenoviral expression of reporter proteins for high-throughput cell-based screening. AB - Recombinant adenoviruses are versatile tools for gene delivery into mammalian tissue-culture cells. Adenoviruses can infect both dividing and nondividing cells, with efficiencies approaching 100%. Here we describe the use of adenoviruses to express reporter proteins in high-throughput cell-based assays. PMID- 16353378 TI - Peptide and small-molecule microarrays. AB - Two methods that use chemoselective ligation chemistry to prepare peptide and small-molecule microarrays are described here. The first method involves the functionalization of a glass slide with a glyoxylyl group, followed by chemoselective ligation of small molecules or peptides to the functionalized surface via a covalent bond. In the second method, peptides or small molecules are first conjugated to a macromolecular scaffold. The final ligand-scaffold conjugates are then spotted and adsorbed onto the solid surface. Three different assay methods to screen such chemical microarrays are described. PMID- 16353379 TI - Development of a yeast two-hybrid screen for selection of human Ras-Raf protein interaction inhibitors. AB - A yeast two-hybrid screening system was developed to screen for small molecules that inhibit the interaction of the Ras and the Raf proteins. Hyperpermeable yeast strains useful for high-throughput screening (HTS) for the two-hybrid system were created. Differential inhibition of the Ras-Raf vs the hsRPB4-hsRPB7 interaction allowed the identification of selective inhibitors. PMID- 16353380 TI - Population-based study of the geographic variation in colon cancer incidence in Alabama: relationship to socioeconomic status indicators and physician density. AB - OBJECTIVES: The objective of this population-based study was to examine the relationship between race, socioeconomic characteristics (socioeconomic status, SES), physician density, and colon cancer incidence in Alabama. METHODS: Data for 5,788 colon cancer cases from 1996 to 1999 provided by the Alabama Statewide Cancer Registry are linked to county-level measures of SES, including median household income, percentage of high school graduates, percentage of families below poverty level, and occupational and health care factors. Poisson regression is used to model the predictors adjusting for age, gender, and race. RESULTS: Blacks had higher incidences of colon cancer compared with whites and presented with later stages (20.4% versus 14.8% for distant disease (P = 0.0089). After controlling for race, gender, and age at diagnosis, significant associations were detected between colon cancer incidence and higher education (RR = 1.10; 95% CI, 1.03-1.17), and increased number of physicians per 1,000 (RR = 1.14; 95% CI, 1.06 1.22). The county percentage of families below poverty is associated inversely with localized disease and positively with distant stage. CONCLUSIONS: Colon cancer incidence varies geographically across Alabama and is positively related to aggregate SES factors, including education and physician density. A higher incidence of distant disease is related to black race and increased poverty. Health disparities in colon cancer across Alabama warrant further investigation. PMID- 16353382 TI - Music and life. PMID- 16353381 TI - PTH (1-34): a novel anabolic drug for the treatment of osteoporosis. AB - The management of osteoporosis has in the past included providing patients with an adequate source of calcium and vitamin D and consideration of some type of antiresorption therapy such as hormone replacement, SERM therapy, or bisphosphonate therapy. However, patients who have very low bone density and continue to have fractures while receiving these therapies need to consider being treated with the recently approved novel anabolic drug PTH (1-34) (Forteo). The osteoblast has the receptor for parathyroid hormone (PTH). When PTH (1 -4) is given in a pulsatile fashion, for example a daily subcutaneous injection, it stimulates osteoblasts to make new bone. PTH (1-34) has been shown to markedly increase spinal bone density and increase hip bone density. In addition, the relative risk of having moderate vertebral fracture was reduced by 90%. Thus, PTH (1-34) offers an exciting new approach for the treatment of severe osteoporosis. PMID- 16353383 TI - The outpatient conundrum. PMID- 16353385 TI - Inflammatory bowel disease update. PMID- 16353384 TI - Malignant melanoma of the auricle. AB - Malignant melanomas are found in a variety of locations, ranging from sun-exposed skin to the nasal cavity and paranasal sinuses. Melanomas arising in the head and neck region comprise some 20% of all melanomas; of these, 7-15% occur in the vicinity of the external ear (most often on the helix). Auricular melanomas, like those arising elsewhere, are rare in childhood and are most often first diagnosed in men in their 50s. Melanomas of the external ear may present as flat pigmented lesions or as raised mass lesions; amelanotic (non-pigmented) variants exist as well. Auricular melanomas are frequently recognized pathologically as either superficial spreading melanomas or nodular melanomas. These tumors are aggressive, with a propensity for spreading to both regional lymph nodes and distant sites. Key pathologic prognostic features of auricular melanomas include the histological subtype, tumor thickness, level of invasion and presence of ulceration. Therapy includes both aggressive surgical attempts at excision in combination with sentinel node sampling in some instances and perhaps, adjuvant therapy as well. This is a tumor which is often overlooked until late in its course, with tragic consequences; vigilance and aggressive attempts at identifying these tumors at earlier stages are strongly advocated. PMID- 16353386 TI - Section F-314 through a long-term care surveyor's eyes. PMID- 16353387 TI - Medicare payment system for wound care dressings. PMID- 16353388 TI - Different effects of intracochlear sensory and neuronal injury stimulation on expression of synaptic N-methyl-D-aspartate receptors in the auditory cortex of rats in vivo. AB - CONCLUSIONS: The expression of synaptic N-methyl-D-aspartate (NMDA) receptors in the auditory cortex is dynamic and is bidirectionally regulated by auditory activity. Furthermore, the time course of changes in the level of NR2A protein differs after sensory and neuronal injury stimulation, which modulate different changes in synaptic plasticity. OBJECTIVE: To examine the effects of different types of auditory activity on the expression of synaptic NMDA receptors (NMDARs) in the auditory cortex of rats. MATERIAL AND METHODS: We prepared synaptosomes from the auditory cortices of postnatal Day 28 ototoxic-deafened Sprague-Dawley rats and postnatal Day 28 Sprague-Dawley rats subjected to noise trauma that were given various treatments and compared them to the synaptosomes of 1-6-week-old normal Sprague-Dawley rats. The expression of different NMDAR subunits in the synaptosomes was investigated by means of Western blotting. RESULTS: Changes in NR1 and NR2B proteins were not significant during different types of auditory activity. The level of NR2A protein increased remarkably during postnatal development and as a result of electrical intracochlear stimulation, auditory deprivation and noise trauma. Seventy-two h after a 2-h period of sensory electrical intracochlear stimulation, the expression of NR2A protein returned to the level caused by auditory deprivation. Seventy-two h after a 3-h period of noise trauma, elevation of the level of NR2A protein was unchanged. We also confirmed that elevation of the level of synaptic NR2A protein was sensitive to protein synthesis inhibitor and NMDAR antagonist. However, transcription inhibitor had no effect on NR2A protein expression. PMID- 16353389 TI - Telemedicine, home care, and reimbursement: legal considerations. PMID- 16353390 TI - "Reasonable and necessary": some case management considerations. PMID- 16353391 TI - Expression of vascular endothelial growth factor and its receptors in the cochlea of various experimental animals. AB - CONCLUSION: The results of this study indicate that vascular endothelial growth factor (VEGF) may be an important regulator of the vascular network of the inner ear and suggest that the VEGF signalling pathway may play a role in pathophysiologic conditions. OBJECTIVE: In order to clarify the role of vascular growth factor in the modulation of the vascular network of the cochlea, we studied the expression of VEGF and its receptors-fms-like tyrosine kinase (Flt-1) and foetal liver kinase (Flk-1)-in the inner ear of 3-month-old rodents of different species: C57BL/6J mice, Wistar albino rats and Hartley albino guinea pigs. MATERIAL AND METHODS: Qualitative immunohistochemical studies were performed by using specific antibodies to VEGF and its receptors on paraffin sections of the cochlea. The expression levels of VEGF and its receptors were quantified by means of Western blot analysis of cochlea protein extracts. RESULTS: We demonstrated that VEGF and its receptors are expressed in the cochlea and described their distribution in the inner ear. In particular, VEGF and Flt-1 are present at the level of the modiolus, spiral ganglion, spiral ligament, basilar membrane, supporting cells, outer and inner hair cells and stria vascularis. Flk-1 was less strongly expressed in the cochlea and was not detected in the organ of Corti. PMID- 16353392 TI - [Functional evolution of bacteria in degradation of environmental pollutants]. PMID- 16353393 TI - [In times of racialization: the case of the 'health of the black population' in Brazil]. AB - The article analyzes initiatives aimed at creating a field of reflection and political intervention called the 'health of the black population,' which occurred between 1996 and 2004, that is, under the administration of Fernando Henrique Cardoso and part of Luis Inacio Lula da Silva's administration. During this period, the process of discussing and enacting affirmative action policies in Brazil gained greater visibility, especially following the UN-sponsored Third World Conference on Racism, Racial Discrimination, Xenophobia, and Related Intolerance (Durban, South Africa, September, 2001). The article describes the emergence of a proposal of compensatory policy within the Brazilian public health system. It then addresses the contemporary debate on race and health, especially the U.S. biomedical literature, and explores how this discussion has been appropriated by agencies and agents concerned with drawing up a 'racial policy' for the public health sector in Brazil. PMID- 16353394 TI - Trans-tympanic silicone plug insertion for chronic patulous Eustachian tube. AB - CONCLUSION: Trans-tympanic insertion of a new silicone plug seems to be useful for controlling the distressing symptoms of patients with a chronic patulous Eustachian tube (PET). OBJECTIVE: To evaluate the effectiveness of a new silicone plug for blocking the isthmus of a PET in patients whose symptoms were resistant to other therapies for > 6 months. MATERIAL AND METHODS: The silicone plug (total length 23-25 mm; tip diameter 1.0-2.0 mm) was inserted in 44 ears of 37 patients with chronic PET. It was inserted through the tympanic orifice of the ET to obstruct the isthmus of the tube via an incision in the anterosuperior portion of the tympanic membrane. RESULTS: Insertion of the plug was possible in all except two ears, in which it failed because of a narrow tympanic orifice of the ET. In 11 ears of 10 patients, the plug was replaced by a larger one using the same approach to improve efficacy. Of the 42 ears in which the silicone plug was successfully inserted, 30 (71.4%) achieved relief from symptoms of PET without additional treatment. In > 60% of these cases, the symptoms of PET were well controlled with an aerated middle ear. The follow-up period ranged from 6 to 68 months (mean 38.9 months). PMID- 16353395 TI - Effect of i.v. dexamethasone on postoperative dizziness, nausea and pain during canal wall-up mastoidectomy. AB - CONCLUSION: Prophylactic i.v. administration of 10 mg of dexamethasone reduced postoperative dizziness and nausea without adverse effects in patients undergoing canal wall-up mastoidectomy, but did not reduce postoperative pain. OBJECTIVE: To investigate the efficacy of dexamethasone in reducing postoperative dizziness, nausea and pain in patients undergoing canal wall-up mastoidectomy. MATERIAL AND METHODS: A prospective, randomized, double-blind study was performed between January 2002 and December 2004. A total of 162 patients (66 males, 96 females) scheduled for canal wall-up mastoidectomy were enrolled and randomly assigned to receive either i.v. dexamethasone (10 mg in 2 ml) or i.v. placebo (2 ml of normal saline) during mastoidectomy. A standard general anesthetic technique was employed throughout the surgical procedure. At 3, 6 and 24 h after surgery, patients completed questionnaires regarding postoperative dizziness, nausea and pain, with their responses being given on a visual analog scale. RESULTS: There were no postoperative complications in either group. Compared to the placebo group, the dexamethasone group showed reduced postoperative dizziness and reduced nausea (p<0.05 for both) at 24 h postoperatively. Compared to the placebo group, dexamethasone had no effect on postoperative pain. PMID- 16353396 TI - Intratympanic gentamicin treatment after endolymphatic sac surgery. AB - CONCLUSIONS: Interval treatment with up to three intratympanic gentamicin injections once weekly effectively controlled vertigo while preserving hearing in patients with Meniere's disease and recurrent or resistant vertigo after saccotomy. OBJECTIVES: Recurrent or resistant incapacitating vertigo may occur after endolymphatic sac surgery (saccotomy) in patients with Meniere's disease. In these patients, revision saccotomy, vestibular nerve section or labyrinthectomy are the established treatment options. We advocate a once-weekly application of intratympanic gentamicin (12 mg) as an effective alternative in this group of patients. MATERIAL AND METHODS: Five patients (age range 39-65 years) with definite Meniere's disease according to the 1995 American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) criteria and incapacitating vertigo underwent gentamicin treatment after saccotomy. Control of vertigo and hearing preservation were the aims of treatment. The follow-up period ranged from 26 to 59 months. History and pure-tone audiometry were used to assess vertigo control and hearing, respectively. The frequency of vertigo in the 6-month period before gentamicin treatment ranged between 0.5 and four definitive episodes per month. Hearing stage (AAO-HNS criteria) before gentamicin treatment ranged between 2 and 4. Pre- and post-treatment pure-tone hearing thresholds at 0.5, 1, 2 and 3 kHz were compared by means of the Mann-Whitney U-test. RESULTS: Complete vertigo control (class A; AAO-HNS) and hearing preservation at 0.5, 1, 2 and 3 kHz were achieved. PMID- 16353397 TI - Validity and reliability study of three tinnitus self-assessment scales: loudness, annoyance and change. AB - CONCLUSIONS: The three tinnitus self-rating scales described herein can be employed as part of "minimal datasets" to reflect the patient's current tinnitus status. These tests are simple and easy to use and can be completed by the patient alone. The results are easy to interpret and provide a good foundation for an effective doctor-patient dialogue. OBJECTIVE: To investigate the reliability and validity of three tinnitus self-rating scales: a six-point response scale for tinnitus loudness; an eight-point response scale for tinnitus annoyance; and a six-point response scale for tinnitus change. MATERIAL AND METHODS: The data for 273 patients participating in 2 separate studies were assessed in terms of their validity and reliability. We used criterion validity to determine whether the scales had empirical associations with external criteria, in this case an already firmly established tinnitus questionnaire. In addition we examined construct validity, i.e. its subcategories convergent and discriminant validity, in order to find out how related or unrelated items or scales were. We tested the reliability and repeatability of the scales using patients on our waiting list for tinnitus desensitization. RESULTS: The test retest reliability was 0.72 for tinnitus loudness and 0.62 for tinnitus annoyance. Calculations showed that all three scales correlated positively with validated complex scales and thus we considered convergent validity to be adequate. PMID- 16353399 TI - Development and characterization of an animal model of dental sinusitis. AB - CONCLUSION: The results of this study confirm that the present rabbit model of dental maxillary sinusitis (dMxS) is reproducible and simulates human dental sinusitis with respect to initiation, progression and inflammation. It is applicable to further studies of sinusitis of odontogenic origin. OBJECTIVES: To induce acute dMxS in rabbits by using their own oral microflora to create a periapical infection and to follow morphological, radiographic, bacteriological and histological changes to the sinus mucosa. MATERIAL AND METHODS: The experimental animals comprised 26 New Zealand White rabbits. Maxillary premolar root canals were identified bilaterally and the continuously growing germs of the roots were severed by diathermy. The animals were randomized into 2 groups: in Group 1 (n=20) the teeth were left open for the entire study period; in Group 2 (n=6) the root canals were sealed 1 week after the initial intervention. The animals in Group 1 were sacrificed at intervals ranging from 2 h to 9 months after intervention. All animals in Group 2 were sacrificed 6 months after intervention. After macroscopic and radiographic examination, post-mortem inspection of the paranasal sinus cavity and maxillary complex and microbiological sampling, the entire nasal sinus complex with the hard palate in situ was resected and processed for serial coronal sectioning. RESULTS: In Group 1, after 3 months, the radiographic changes ranged from widening of the periodontal space to bone reaction. At sacrifice, changes in the sinus mucosa ranged from signs of mucosal inflammation to purulent dMxS. Microbial growth, predominantly Gram-negative aerobes, increased over time. In Group 2, the findings were generally more pronounced. Anaerobic microorganisms were predominant. In both groups the findings were consistent with dMxS. PMID- 16353398 TI - A mutation in Wolfram syndrome type 1 gene in a Japanese family with autosomal dominant low-frequency sensorineural hearing loss. AB - CONCLUSION: Our findings suggest that Wolfram syndrome type 1 gene (WFS1) mutation is an important cause of autosomal dominant low-frequency sensorineural hearing loss (LFSNHL) in Japan. OBJECTIVE: DFNA6/14 is caused by a heterozygous mutation of WFS1 and is a common cause of autosomal dominant LFSNHL among populations in both Europe and the US. The purpose of this study was to investigate WFS1 mutations among Japanese patients whose phenotypes were consistent with those of DFNA6/14. MATERIAL AND METHODS: Using audiometry and genetic analysis, we searched for WFS1 mutations in three unrelated Japanese patients with LFSNHL and a familial history of autosomal dominant hearing loss. RESULTS: One patient carried a heterozygous G2700A mutation at codon 844 in exon 8, resulting in substitution of a threonine for an alanine (A844T). Genetic analysis of the available members of the patient's family showed that the A844T mutation segregated with LFSNHL, but was not detected in any of 140 control chromosomes. It thus appears likely that the A844T mutation is causative for hearing loss in this group. Speech audiometry, self-recording audiometry and auditory brainstem responses showed the patient to have cochlear deafness without retrocochlear dysfunction. No mutation was found in the other two patients. PMID- 16353400 TI - Organic first: a biology-friendly chemistry curriculum. PMID- 16353401 TI - Using long-term time-lapse imaging of mammalian cell cycle progression for laboratory instruction and analysis. PMID- 16353402 TI - Taste disorders: a survey of the examination methods and treatments used in Japan. AB - CONCLUSIONS: In Japan, the number of patients seeking treatment for taste disorders from otolaryngologists is approximately 240 000/year, and this number has almost doubled in the last 13 years. Taste examinations are routinely conducted in almost all university hospitals; testing is not as common in private clinics. Administration of zinc is the main treatment for taste disorders in Japan. OBJECTIVES: To investigate the number of patients who visited otolaryngologists for treatment of taste disorders between 2000 and 2002. In addition to determining changes in the number of patients that had occurred in the 13 years since the last study in 1990, the survey also sought to find out what types of taste examination were used to diagnose taste disorders in Japan and what therapies were used to treat them. MATERIAL AND METHODS: A questionnaire was mailed to 1559 members of the Japan Society of Stomato-pharyngology. RESULTS: The average number of patients with taste disorders who visited otolaryngological clinics between 2000 and 2002 was 244 858/year. This number is 1.77-fold greater than the number of patients reported by the last study in 1990 (138 575). Electrogustometry was used in 99% of university hospitals and in 20% of private clinics. The filter-paper disk method was also used in 57% of university hospitals and in 27% of private clinics. The main treatment was zinc administration. Most doctors (70%) prescribed polaprezinc, a zinc-containing drug that is used to treat peptic ulcers. Of these doctors, 78% stated that it effectively treated taste disorders and 11% that it did not. PMID- 16353403 TI - Issues raised by prophylactic vaccine for cervical cancer. PMID- 16353404 TI - Carcinogenicity of polycyclic aromatic hydrocarbons. PMID- 16353405 TI - Occupational rhinitis: diagnostic criteria, relation to lower airway symptoms and IgE sensitization in bakery workers. AB - CONCLUSIONS: The use of different diagnostic criteria has considerable consequences for the prevalence estimates of occupational rhinitis. There is a strong relationship between occupational rhinitis and lower airway symptoms. Storage mites appear to be important occupational allergens in Norwegian bakeries. OBJECTIVES: To study the consequences of various diagnostic criteria on the prevalence of occupational rhinitis, assess the prevalence of IgE sensitization and explore the relationships between upper and lower airway symptoms and between symptoms and IgE sensitization. MATERIAL AND METHODS: A total of 197 employees in 6 bakeries were interviewed and completed a questionnaire. A skin prick test was performed, total and specific IgE were determined and a histamine release test was performed for relevant allergens. The criteria for the diagnosis of occupational rhinitis were based on the 1994 International Consensus Report on Rhinitis. RESULTS: The prevalence of occupational rhinitis varied between 23% and 50%, depending on the criteria used. The occurrence of nasal symptoms was found to precede the development of lower airway symptoms. Occupational rhinitis, both IgE- and non-IgE-mediated, was associated with asthma symptoms. The most frequent causes of sensitization (20%) were different species of storage mites. Storage mite sensitization was related to occupational rhinitis and work exposure. PMID- 16353406 TI - Survivin expression is significantly higher in pN+ oral and oropharyngeal primary squamous cell carcinomas than in pN0 carcinomas. AB - CONCLUSIONS: The preliminary results reported here suggest that survivin expression in primary oral and oropharyngeal squamous cell carcinomas (SCCs) may identify patients at risk of disease disseminating to neck lymph nodes. If these results are confirmed in larger series of patients it may imply that elective neck dissection should be considered in clinically N0 patients with oral and oropharyngeal SCCs who show high expression of survivin. OBJECTIVE: To investigate the expression of survivin, a member of the inhibitor of apoptosis proteins family, in patients with primary oral and oropharyngeal SCCs with and without neck lymph node metastases. MATERIAL AND METHODS: We considered 13 consecutive cases of oral and oropharyngeal SCCs with lymph node metastases (pN + ) and 13 cases of pN0 oral and oropharyngeal SCCs. The survivin reactivity of primary SCCs and lymph node metastases was evaluated immunohistochemically. A lesion was considered positive if >9.5% of the tumour cells showed diffuse strong staining. RESULTS: Sporadic groups of normal basal and parabasal epithelial cells showed weak survivin staining. In SCCs, a nuclear reaction predominated. Eight primary pN+ SCCs were survivin-positive (mean expression 34.7%), compared to 5 primary pN0 SCCs (mean expression 12.3%; p=0.017). Statistical analysis disclosed significantly higher survivin expression in primary oral and oropharyngeal SCCs that developed distant non-lymphatic metastases (p=0.012). PMID- 16353407 TI - Whole-body MRI for staging patients with head and neck squamous cell carcinoma. AB - CONCLUSIONS: Whole-body MRI is feasible for the tumor staging of patients with malignant head and neck tumors and appears to be a quick, reliable and proven alternative in general and for patients with contraindications to CT. This examination minimizes the logistical effort required compared to multimodality strategies. Its economic impact remains to be determined. OBJECTIVE: To assess the performance of whole-body MRI for staging patients with squamous cell carcinoma of the head and neck region. MATERIAL AND METHODS: This was a randomized, prospective clinical study. For tumor staging, 21 patients (mean age 56.7 years; range 43-80 years) with advanced malignant head and neck tumors underwent whole-body MRI in addition to routinely performed imaging investigations, including sonography, chest X-ray, CT of the head, neck and thorax and endoscopy. All investigations were accomplished within a period of 10+/-3 days in a random order. A randomized, blinded, consensus assessment of all the whole-body MRI examinations was performed by two radiologists. The localization and extent of the primary tumor and metastases were documented for whole-body MRI and compared to the standard of reference (all other imaging modalities as well as histology). Point estimates of the diagnostic accuracy of whole-body MRI were calculated. RESULTS: In accordance with the standard of reference, the overall TNM category was correctly determined with whole-body MRI in all 21 patients. However, four patients were classified as having carcinoma of unknown primary, as the primary tumor was not found with any imaging modality. Two patients had mediastinal, pulmonary and hepatic metastases. PMID- 16353408 TI - Delayed otogenic hydrocephalus after acute otitis media in pediatric patients: the changing presentation of a serious otologic complication. AB - CONCLUSIONS: The clinical presentation of otogenic dural sinus thrombosis (DST) as a complication of acute otitis media (AOM) can be masked by antibiotic treatment. Morning episodes of vomiting and/or headache, visual impairment and a history of AOM seem to be indicative of otogenic hydrocephalus. We therefore advocate that the MRI scans of patients with similar symptoms should be carefully studied to facilitate the early diagnosis of a potentially life-threatening complication. OBJECTIVE: To describe the frequency, pathognomonic signs, clinical course and outcome of otogenic hydrocephalus and DST as complications of AOM in pediatric patients. MATERIAL AND METHODS: We undertook a retrospective chart review of all pediatric patients (age 1-14 years) treated for otitis media and its complications at an academic medical center between 1999 and 2003. The main outcome measures were otologic and ophthalmologic findings and CT and MRI scans at the beginning of treatment and 3 months later. RESULTS: We report on five cases with otogenic DST following AOM. All but one of them presented initially with diplopia caused by otogenic hydrocephalus. In four cases the otologic complaints had already disappeared by the time of MRI confirmation of the diagnosis. Only one child was referred with severe otologic symptoms. Management included systemic antibiotics, short-term heparin anticoagulation and surgical decompression. In our cases, even after intensive i.v. antibiotic treatment, only surgery led to a significant improvement in the clinical condition. PMID- 16353409 TI - Primary tuberculous petrositis. AB - Tuberculous osteomyelitis of the temporal bone is a rare and dangerous entity that should be included in the differential diagnosis of infectious processes of the base of the skull. We present the case of an 11-year-old child who presented with diplopia, ear discharge and hearing loss. The radiological and histopathological findings revealed tuberculous otitis with osteomyelitis and an abscess in the petrous apex. The child responded to anti-tuberculous chemotherapy. The diagnosis and management of tuberculous osteomyelitis are discussed and a brief review of the literature is presented. PMID- 16353410 TI - Hereditary angioneurotic edema of the larynx. AB - Hereditary angioneurotic edema (HAE) is an autosomal dominant disease resulting from a deficiency of functional C1-esterase inhibitor. If not recognized promptly and treated properly the disease can result in a fatal outcome as it causes laryngeal edema, which can lead to a life-threatening acute upper airway obstruction. We present the case of a 37-year-old female with HAE of the larynx, who was diagnosed early and treated properly, together with a review of the literature. PMID- 16353411 TI - Zero tolerance of violence. PMID- 16353412 TI - Epidemiological approach to predicting psychiatric risk in the military. PMID- 16353413 TI - Is gestational week at birth a predictor of schizophrenia? PMID- 16353414 TI - Stigmatisation of people with schizophrenia in Japan. PMID- 16353415 TI - Surgical-site infection: still a reality. PMID- 16353416 TI - Tighten your glycemic control. PMID- 16353417 TI - Extreme makeover--OR edition. PMID- 16353418 TI - Slash sharps risk for surgical personnel. PMID- 16353419 TI - Aphonia and dysphagia after gastrectomy. AB - A 67-year-old male was referred to our otolaryngological clinic because of aphonia and dysphagia. His voice was breathy and he could not even swallow saliva following a total gastrectomy for gastric carcinoma performed 2 weeks previously. Laryngeal fiberscopy revealed major glottal incompetence when he tried to phonate. However, both vocal folds abducted over the full range during inhalation. The patient could not swallow saliva because of a huge glottal chink, even during phonation. Based on these findings, he was diagnosed as having bilateral incomplete cricoarytenoid dislocation after intubation. The patient underwent speech therapy; within 1 min his vocal fold movement recovered dramatically and he was able to phonate and swallow. There have been few case reports of bilateral cricoarytenoid dislocation, and no effective rehabilitation has been reported. We believe that our method of vocal rehabilitation serves as a useful reference for physicians and surgeons worldwide. PMID- 16353420 TI - Diagnostic criteria for migrainous vertigo. PMID- 16353421 TI - Galectins: novel anti-inflammatory drug targets. AB - Galectins are a protein family defined by their affinity for beta-galactosides and consensus sequences. They are pleiotropic regulators involved in a multitude of functions, both in and out of the cell. Extracellularly, they have the potential to bind to various surface receptors on a variety of cell types as well as extracellular matrix (ECM) proteins, thus causing cell activation or apoptosis, modulating cell adhesion and inducing cell migration. Intracellularly, they can regulate cell growth, apoptosis and cell cycle progression. Galectins are either pro-inflammatory or anti-inflammatory. Some, such as galectin-1, may be employed as anti-inflammatory agents, while others, such as galectin-3, are evidently suitable targets for anti-inflammatory drugs. The extracellular functions of galectins involve their lectin-carbohydrate interactions and thus their carbohydrate ligands or mimetics would be suitable inhibitors. While the intracellular functions of galectins do not appear to engage lectin-carbohydrate interactions, the carbohydrate-binding sites of these proteins may still be involved. Therefore, the same inhibitors may be used regardless of whether intracellular or extracellular galectins are to be targeted. PMID- 16353422 TI - Binge eating disorder affects outcome of comprehensive very-low-calorie diet treatment. AB - To determine the effects of binge eating disorder (BED) on weight loss and maintenance in women undergoing treatment for obesity, we studied the weight changes of 38 women (body mass index > 30 kg/m2), 21 of whom met proposed criteria for BED and 17 of whom reported few problems with binge eating, during and after a 26-week comprehensive very-low-calorie diet (VLCD) treatment program. All 17 subjects without and 16/21 subjects with BED returned for four follow-up visits over 12 months (p = 0.05). While a similar proportion of subjects with and without BED reported absolute adherence to both the modified fast and refeeding, those with BED showed a significantly different distribution in energy intake from those without BED, with fewer small and more large lapses among those who deviated from the diet (p < 0.05). There was no significant difference in mean weight loss over the 26 weeks of treatment, but subjects with BED showed significantly diminished weight loss during the middle third of treatment (p < 0.05). Black subjects, regardless of the presence of BED, lost significantly less weight during treatment than white subjects (p < 0.005). Although there was no significant difference in mean weight loss at any of the four follow-up visits between subjects with and without BED, 25% of subjects with BED had regained > 50% of their lost weight by three-month follow-up, vs. no subjects without the disorder (p < 0.05). One year after completing treatment, approximately half of BED (+) and BED (-) subjects had a good outcome, maintaining a weight loss > or = 10% of initial body weight. However, 35% of subjects with BED, and none of the subjects without BED, had a poor outcome (p < 0.05). We conclude that many individuals with BED will respond well to a medically supervised comprehensive VLCD program, attaining medically significant weight loss. However, this subgroup appears to be at risk for early major regain of lost weight and for poor outcome one year following weight-loss treatment. PMID- 16353423 TI - Factors associated with weight control in older adults. AB - Dieting behaviors in a sample of 183 overweight older adults were studied to assess how they were influenced by six cognitive, behavioral, emotional, and social variables. Membership in a weight control program was also evaluated to assess whether it affected these relationships. Responses indicated that reports of high quality dieting behaviors were associated with higher levels of depression and less effective coping skills. Dieting behaviors among subjects who were participants in weight loss programs were not as strongly associated with less effective coping skills, but were associated with external health locus-of control. The degree of social support had a limited impact on dieting behaviors, while measures of optimism and health status were unrelated to dieting behaviors. We concluded that older adults, especially those who diet independently, are likely to experience significant stress associated with weight loss efforts. Weight loss programs for older adults might produce better outcomes if they focus on reducing depression and stress associated with dieting. PMID- 16353424 TI - Comparison of cardiovascular risk factors in obese blacks and whites. AB - Blacks are known to have higher blood pressure levels, a higher prevalence of hypertension, and higher body weights than whites. However, the interrelationships of these and other cardiac risk factors have not been analyzed in an obese population. We compared blood pressure (BP) and lipid levels in 174 obese blacks and 939 obese white patients who were entering a weight loss program; we also assessed the effects of weight loss on these factors. Prevalence of treated hypertension was similar in blacks and whites (28% vs. 25%, respectively). In patients not taking BP medication, black women weighed more (108 kg) than white women (102 kg) and black and white males' weights were similar (135 kg vs. 131 kg). Systolic and diastolic BP were similar in black and white women; black males had similar SBP but a significantly lower DBP than white males (83 mmHg vs. 89 mmHg, respectively). Lipid levels were similar in black and white women except black women had lower triglycerides (1.30 mmol/L) than white women (1.58 mmol/L, p < 0.05); and black males compared to white males had significantly lower total cholesterol (4.76 mmol/L vs. 5.56 mmol/L), LDL cholesterol (3.15 mmol/L vs. 3.52 mmol/L) and triglycerides (1.31 mmol/L vs. 2.17 mmol/L, p < 0.05). Adult-onset obesity adversely affected a number of cardiovascular risk factors in whites, but not in blacks. Blacks lost significantly less weight (-13 kg) than whites (-19 kg). However, controlling for the difference in weight loss, blacks sustained comparable improvement in lipids and blood pressure, except for TC/HDL-C (whites improved significantly more, 0.36 kg/m2, than blacks, 0.03 kg/m2). Thus, the impact of obesity on cardiovascular risk factors seems ameliorated in blacks compared to whites. PMID- 16353425 TI - Influence of opioids in hypothalamic nuclei on cold thermogenesis of lean and obese LA/N-cp rats. AB - An overactive endogenous opioid peptide system (EOP) in the hypothalamus of the obese rats could contribute to a subnormal metabolic response to cold stress. Specific mu, delta, kappa opioid receptor antagonists and naloxone were infused into cannulaes aimed at the paraventricular nucleus (PVN) of awake freely moving obese (LA/N-cp corpulent) and lean littermate rats. Metabolic responses were measured by indirect calorimetry during thermoneutrality (30 degrees C) and at 10 degrees C for 60 minutes each. When expressed relative to metabolic body size (kg(-.75)) obese rats had lower values (obese = 21.1 +/- 1.9 vs. lean = 27.9 +/- 2.7 ml x kg(-.75) x min, mean +/- s.d., p < 0.05) at 10 degrees C during saline infusion. EOP antagonist infusions at 30 degrees C had no effect on metabolic rate for either lean or obese animals. Mu (23.5 +/- 3.4 ml x kg x (-75) x min) and delta (23.0 +/- 2.0) antagonism and naloxone (25.0 +/- 2.3) significantly increased the metabolic response to cold in obese but not lean rats. These data suggest that certain subtypes of EOP receptors in or near PVN are overactive in obese rats. This overactive state may inappropriately inhibit the thermogenic response to cold stress in obesity. PMID- 16353426 TI - Stress-induced cortisol response and fat distribution in women. AB - Recent studies have shown an association between uncontrollable stress and abdominal fat distribution. It has been suggested that changes in cortisol secretion might represent one possible mechanism for this relationship. This study investigated whether body fat distribution, determined by waist-to-hip ratio (WHR), is related to salivary cortisol levels in response to laboratory stressors. Subjects were 41 overweight women with a Low or a High WHR. Multiple measures of cortisol and mood were obtained during a session of stressful tasks (eg., timed arithmetic) and during a time-matched, control rest session. Also, background life stress and psychological trait variables were assessed. Compared to Low WHR subjects, High WHR subjects secreted significantly more cortisol during the stressful session after 60 minutes of stress, and considering the total area under the curve of secretion. This difference was not seen on the rest day. In terms of background and psychological measures, High WHR subjects were characterized by poorer coping skills and differences in mood reactivity. Specifically, although all subjects became more angry in response to the stressful session, High WHR subjects showed smaller increases in anger. This could indicate that they are more likely to evidence a helpless reaction to uncontrollable stress. These findings support the hypothesis that cortisol secretion might represent a mechanism for the observed association between stress and abdominal fat distribution. Furthermore, differences in coping and appraisal may suggest that a particular psychological pattern might influence the reactivity of the adrenal-cortical system to stress, and subsequent fat distribution. PMID- 16353427 TI - Growth hormone secretion after testosterone administration to men with visceral obesity. AB - Visceral obesity in men has been reported to be characterized by low testosterone (T) and insulin-like growth factor I (IGF-I) concentrations, the latter suggesting a relative growth hormone (GH) deficiency. Since T and GH-secretions are interrelated, men with visceral obesity were substituted with T for 14 days, and diurnal secretion pattern of GH as well as IGF-I concentrations, and metabolic variables were followed. Visceral obese men were characterized by a decreased total GH secretion and diminished peak amplitude, size, and number. T substitution was followed by elevation of IGF-I levels. The IGF-I increase correlated with the elevation of T-concentration, and was most pronounced in men with the lowest concentrations of free T from the outset. There were no detectable changes in total quantity, amplitude, size or number of peaks of GH secretion. Glucose, chlolesterol and triglycerides as well as diastolic blood pressure decreased. There were no changes in thyroid or hematology variables. T substitution of visceral obese men is followed by an elevation of IGF-I concentrations. It is suggested that this might be due either to minor, non detectable increases in GH secretion, or to direct effects of T on IGF-I concentrations. The regulatory mechanisms by which T-administration are leading to metabolic and anthropometric improvements, might be direct effects of T, with or without mediation via GH secretion. PMID- 16353428 TI - Effects of a fat substitute, sucrose polyester, on food intake, body composition, and serum factors in lean and obese Zucker rats. AB - Sucrose polyester, a fat substitute, has shown promise in reducing blood cholesterol and body weight of obese individuals. Effects of this compound in the Zucker rat, a genetic model of obesity, are unknown. Thus, we examined food intake, body weight, body composition, and several metabolic parameters in sera of lean and obese female Zucker rats. Eight-week-old lean and obese animals were given a choice between a control diet (15% corn oil) and fat substitute diet (5% corn oil and 10% sucrose polyester) for 2 days. Next, one-half of the lean and obese groups received control diet; the remaining lean and obese rats received fat substitute diet for 18 days. Cumulative food intake was depressed in fat substitute groups relative to control-fed animals; however, this effect was more predominant in obese animals. Obese rats consuming fat substitute diet (O-FS) gained less weight as compared to obese control-fed animals (O-C). Lean rats given fat substitute (L-FS) did not have significantly different body weights as compared to the L-C group. Fat substitute groups, combined, had lower body fat and higher body water as compared to controls. The O-FS group had lower serum glucose and insulin and higher fatty acid levels compared to the O-C group. There were no differences in serum cholesterol, HDL, or triglyceride levels due to fat substitute diet. These data suggest that the obese Zucker rat is unable to defend its body weight when dietary fat is replaced with sucrose polyester. PMID- 16353429 TI - Binge eating disorder and the treatment of obesity. PMID- 16353430 TI - Letters to the editor: an editorial. PMID- 16353431 TI - Amphetamine: the Janus of treatment for obesity. PMID- 16353432 TI - Human autonomic pharmacology. XVI. Benzedrine sulfate as an aid in the treatment of obesity. 1938. PMID- 16353433 TI - The striking differences in the prevalence of obesity by race. PMID- 16353434 TI - Validation and application of single breath-hold cine cardiac MR for ventricular function assessment in children with congenital heart disease at rest and during adenosine stress. AB - PURPOSE: To validate the accuracy of ventricular function analysis using a single breath-hold real-time steady sate free precession (SSFP) cine MR method and demonstrate its application during adenosine stress imaging in children with congenital heart disease. MATERIALS AND METHODS: Twenty-eight subjects with congenital heart disease were studied (mean age 12.4 +/- 2.3 years) with MR imaging at 1.5 T. Short-axis images covering the entirety of both ventricles were acquired at rest, with a conventional segmented SSFP cine sequence acquired over multiple breath-holds and a single breath-hold real-time SSFP sequence. Seventeen subjects were given an infusion of adenosine, and the single breath-hold real time short-axis stack was repeated during stress. Two independent observers performed the ventricular function analysis. Data was compared between the 2 acquisition methods at rest and between the single breath-hold acquisition at rest and during adenosine stress. RESULTS: There was good agreement between the multiple breath-hold and single breath-hold methods for measurement of end diastolic volume (r = 0.95 and 0.96, p < .0001) and end-systolic volume (ESV) (r = 0.76 and 0.90, p < .0001) for the left and right ventricles respectively, and the left ventricular mass (r = 0.97, p < .0001). Adenosine was administered safely to all subjects. During stress there were significant changes in the heart rate and ESV, which led to a significant increase in cardiac output (mean 1.5 +/- 1.0 L/min, p < .001). CONCLUSION: Single breath-hold real-time SSFP cine imaging is robust and accurate in assessing cardiac function in children with congenital heart disease. An application of this method is the rapid assessment of cardiac function during adenosine stress. PMID- 16353435 TI - Quantifying regional right ventricular function in tetralogy of Fallot. AB - Right ventricular (RV) function is notoriously difficult to quantify. Patients with tetralogy of Fallot (TOF) have decreased systolic performance. We measure regional RV performance using MRI with 1-dimensional myocardial tissue tagging. By tagging cine-MRI in two views, we measured regional shortening in 12 regions throughout the RV. We image 32 pediatric patients: 21 normal patients and 11 patients with repaired TOF. We establish a normal range for each RV region. TOF patients have decreased shortening on a region-by-region basis. We conclude that regional RV performance can be measured using this technique, and that decreased performance can be demonstrated in TOF patients. PMID- 16353436 TI - Reproducibility and inter-observer variability of dobutamine stress CMR in patients with severe coronary disease: implications for clinical research. AB - PURPOSE: The purpose of this study was to analyze reproducibility and inter observer variability of dobutamine stress cardiac magnetic resonance imaging (dobutamine CMR) and its implications on serial studies. METHODS: Nineteen consecutive patients underwent two dobutamine CMR each (median 12 days apart), as part of eligibility criteria for phase I/II stem cell therapy trial. These patients had Canadian Cardiovascular Society Class III/IV angina despite maximal therapy. The two studies were compared for reproducibility of stress response. To assess inter-observer variability, 29 randomly selected dobutamine CMR studies were analyzed by three experienced observers and Kappa values were computed to measure the agreement. RESULTS: Dobutamine CMR studies were completed without any major complications. The left ventricular function, dobutamine and atropine dose, hemodynamic response, symptomatic response and the results of wall motion and perfusion abnormalities were highly reproducible between the two studies (p = .91). Sample size calculations suggested that a clinical trial using dobutamine CMR to detect an endpoint of resolution of two ischemic segments would require a sample size of 20 subjects and to detect an improvement in perfusion of two segments would require a sample size of 8 subjects. Inter-observer variability between individual and consensus interpretation of dobutamine CMR was good to very good (kappa = 0.81 for wall motion and 0.70 for perfusion). CONCLUSION: Dobutamine CMR is a highly reproducible technique with very good inter-observer variability and could be used as a specific endpoint in a relatively small clinical trial. PMID- 16353437 TI - Modified RV short axis series--a new method for cardiac MRI measurement of right ventricular volumes. AB - PURPOSE: The current standard image orientation employed in the MRI assessment of right ventricular volumes uses a series of short axis cine acquisitions located with respect to a horizontal long axis view with the first slice placed across the atrio-ventricular valve plane at end diastole. Inherent inaccuracies are encountered with the use of this image orientation due to difficulty in defining the tricuspid valve and the border between atrium and ventricle on the resultant images. Our experience indicates that because the tricuspid valve is usually not in-plane in the slice the atrio-ventricular margin is difficult to distinguish. This leads to inaccuracies in measurements at the base of the RV and miscalculation of the RV volume. The purpose of this study was to assess an alternative method of image orientation aimed at increasing the accuracy of RV volume measurements using current commercially available CMRI sequences. This technique, the modified RV short axis series, is oriented to the outflow tract of the right ventricle. METHOD: We undertook a prospective study of 50 post cardiac transplant patients. A series of LV short axis multi-slice cine acquisition FIESTA images was acquired using the current standard technique. From this data set, LV and RV stroke volumes were derived on an Advantage Windows workstation using planimetry of the endocardial and epicardial borders in end systole and end diastole. Our new technique involved obtaining a set of multi-slice cine acquisition FIESTA images in a plane perpendicular to a line from the centre of the pulmonary valve to the apex of the RV. Planimetry of the RV was then performed and a stroke volume calculated using the same method of analysis. RV stroke volumes obtained from both techniques were compared with LV stroke volumes. Three operators independently derived RV data sets. RESULTS: On the images acquired with the new technique, the tricuspid valve was easier to define leading to more accurate and reproducible planimetry of ventricular borders. RV stroke volumes calculated from the new method showed better agreement with LV stroke volumes than with the current method. These results were consistent across the three operators. CONCLUSIONS: This new method improves visualisation of the tricuspid valve and makes analysis easier and less prone to operator error than the current standard technique for MRI assessment of RV volumes. PMID- 16353438 TI - Normal human left and right ventricular and left atrial dimensions using steady state free precession magnetic resonance imaging. AB - PURPOSE: The aim of this project was to establish a database of left and right ventricular and left atrial dimensions in healthy volunteers using steady-state free precession cardiac magnetic resonance imaging, the clinical technique of choice, across a wide age range. METHODS: 108 healthy volunteers (63 male, 45 female) underwent cardiac magnetic resonance imaging using steady-state free precession sequences. Manual analysis was performed by 2 experienced observers. RESULTS: Left and right ventricular volumes and left ventricular mass were larger in males than females: LV end-diastolic volume 160 +/- 29 mL vs. 135 +/- 26 mL, LV end-systolic volume 50 +/- 16 mL vs. 42 +/- 12 mL; RV end-diastolic volume 190 +/- 33 mL vs. 148 +/- 35 mL, RV end-systolic volume 78 +/- 20 mL vs. 56 +/- 18 mL (p < .05 for all). Normalization of values to body surface area removed the statistical differences for LV volumes, but not for LV mass or RV volumes. With increased age, males showed a significant decrease in volume and mass indices for both ventricles, while female values remained unchanged. Compared to females, males had significantly larger maximal left atrial volumes (103 +/- 30 mL vs. 89 +/- 21 mL, p = .01) and left atrial stroke volumes (58 +/- 23 mL vs. 48 +/- 15 mL, p = .01). There was no difference in left atrial ejection fraction between the sexes. CONCLUSION: We have produced a large database of age-related normal ranges for left and right ventricular function and left atrial function in males and females. This will allow accurate interpretation of clinical and research datasets. PMID- 16353439 TI - MR coronary artery imaging with 3D motion adapted gating (MAG) in comparison to a standard prospective navigator technique. AB - Magnetic resonance coronary angiography (MRCA) has been proven to be feasible for imaging of the proximal and medial portions of the three main coronary arteries. Free breathing techniques allow for high resolution imaging but prolong scan time. This could potentially be shortened by improving the efficiency, robustness and accuracy of the navigator gating algorithm. Aim of this study was to determine the feasibility, efficiency, and image quality of a new motion compensation algorithm (3D-MAG) for coronary artery imaging with navigator techniques. In 21 patients the coronaries were imaged in plane with a 3D k-space segmented gradient echo sequence. A T2 preparation prepulse was used for suppression of myocardial signal, during free breathing and a navigator technique with using real time slice following and a gating window of 5 mm was applied to suppress breathing motion artefacts. Imaging was performed with standard gating and compared to 3D-MAG. Image quality was visually compared, contrast-to-noise and signal-to-noise ratio were calculated, the length of visualized coronary arteries was measured and scan duration and scan efficiency were calculated. Standard navigator imaging was feasible in 19 of 21 (90.5%) patients 3D-MAG in 21/21 (100%). Scan efficiency and duration was significantly improved with 3D-MAG (p < .05) without change in image quality. 3D-MAG is superior to conventional navigator correction algorithms. It improves feasibility and scan efficiency without reduction of image quality. This approach should be routinely used for MR coronary artery imaging with navigator techniques. PMID- 16353440 TI - Sample size calculation for clinical trials using magnetic resonance imaging for the quantitative assessment of carotid atherosclerosis. AB - PURPOSE: To provide sample size calculation for the quantitative assessment of carotid atherosclerotic plaque using non-invasive magnetic resonance imaging in multi-center clinical trials. METHODS. As part of a broader double-blind randomized trial of an experimental pharmaceutical agent, 20 asymptomatic placebo control subjects were recruited from 5 clinical sites for a multi-center study. Subjects had 4 scans in 13 weeks on GE 1.5 T scanners, using TOF, T1-/PD-/T2- and contrast-enhanced Tl-weighted images. Measurement variability was assessed by comparing quantitative data from the index carotid artery over the four time points. The wall/outer wall (W/OW) ratio was calculated as wall volume divided by outer wall volume. The percent lipid-rich/necrotic core (%LR/NC) and calcification (%Ca) were measured as a proportion of the vessel wall. For %LR/NC and %Ca, only those subjects that exhibited LR/NC or Ca components were used in the analysis. RESULTS: Measurement error was 5.8% for wall volume, 3.2% for W/OW ratio, 11.1% for %LR/NC volume and 18.6% for %Ca volume. Power analysis based on these values shows that a study with 14 participants in each group could detect a 5% change in W/OW ratio, 10% change in wall volume, and 20% change in %LR/NC volume (power = 80%, p < .05). The calculated measurement errors presume any true biological changes were negligible over the 3 months that subjects received placebo. CONCLUSION: In vivo MRI is capable of quantifying plaque volume and plaque composition, such as %lipid-rich/necrotic core and %calcification, in the clinical setting of a multi-center trial with low inter-scan variability. This study provides the basis for sample size calculation of future MRI trials. PMID- 16353441 TI - MR coronary angiography using 3D-SSFP with and without contrast application. AB - We compared the performance of a contrast-enhanced with a non-contrast breath hold 3D-SSFP-sequence for Magnetic Resonance Coronary Angiography in seven healthy subjects and 14 patients. Visibility of coronary segments, vessel length, image quality and the influence of an extracellular contrast agent (Gadolinium DTPA) were assessed. Overall, the performance of the sequence was better in healthy subjects than in patients. Although the application of Gadolinium-DTPA increased the contrast-to-noise-ratio of the right coronary artery, the overall performance was not significantly improved. We conclude that a 3D-SSFP-technique depicts extensive parts of the coronary arteries and does not require contrast application. PMID- 16353442 TI - Interstudy reproducibility of quantitative perfusion cardiovascular magnetic resonance. AB - PURPOSE: To determine the interstudy reproducibility of quantitative first-pass perfusion cardiovascular magnetic resonance with comparison of 2 previously described analysis techniques. There is no published data on the interstudy reproducibility of perfusion cardiovascular magnetic resonance which can be used to determine the significance of longitudinal changes in myocardial perfusion after pharmacologic or therapeutic interventions with defined sample sizes. METHODS: Sixteen subjects (7 normal volunteers, 9 patients with coronary artery disease) had rest and adenosine stress perfusion cardiovascular magnetic resonance studies on two separate visits. A short axis slice was studied on each visit using a fast low-angle shot sequence. The global and regional myocardial perfusion reserve indices were calculated using 2 methods: model based constrained deconvolution with the Fermi function, and normalized upslopes. Reproducibility was defined as the standard deviation of the measurement differences, divided by the mean (coefficient of variation). RESULTS: The reproducibility of global myocardial perfusion reserve indices was 21% in normal volunteers, which was similar to that in patients with coronary artery disease (CAD) (23%, p = .88). The reproducibility of regional myocardial perfusion reserve indices was 28% (p = .45 vs. global analysis). The reproducibility of global MPRi was superior with Fermi deconvolution compared with normalized upslopes (21% vs. 41%, p = .02). CONCLUSION: At this stage of clinical development, the reproducibility of quantitative perfusion cardiovascular magnetic resonance is good, and superior using Fermi deconvolution in preference to upslope analysis. PMID- 16353443 TI - Cardiomyopathy in Becker muscular dystrophy--does regional fibrosis mimic infarction? AB - We present a case of a 39-year-old man with Becker muscular dystrophy and severe congestive cardiac failure. Cardiac magnetic resonance imaging revealed subendocardial late gadolinium enhancement, similar to that seen in myocardial infarction. He had no risk factors for atherosclerotic coronary artery disease and coronary angiography was normal. We propose that regional subendocardial myocardial fibrosis which has been described histologically in the cardiomyopathy associated with Becker muscular dystrophy may resemble previous infarction at contrast enhanced cardiac magnetic resonance imaging. PMID- 16353444 TI - Magnetic resonance criteria for future trials of cardiac resynchronization therapy. AB - Current patient selection criteria for Cardiac Resynchronization Therapy (CRT), an efficacious treatment for heart failure, include no measure of disconjugate cardiac contractility other than prolonged QRS on electrocardiogram. Using cardiac magnetic resonance imaging, we examined the roles of cardiac asymmetry, asynchrony, and circumferential strain in DCC with the principal aim of generating a robust numerical index for use in future trials of CRT. Standard cardiac magnetic resonance imaging was done on a GE 1.5 Tesla Signa LX MRI clinical scanner (GE Healthcare, Milwaukee, WI, USA) and analyzed by MASS Analysis (MEDIS, Leiden, The Netherlands). The methods were evaluated in eleven patients with advanced heart failure due to ischemic and non-ischemic cardiomyopathy, who did not qualify under current criteria for CRT, five CRT candidates pre-op and eleven normal subjects. Using t-test and standardized differences (SD = sd/diff, Power (N) = number of patients to reach p < .05) we determined efficacy. Indices of asymmetry and asynchrony (Ism and Isn, respectively) could be measured with accuracy and provided excellent statistical power when used as surrogate markers to delineate heart failure and CRT patients from control subjects. Asymmetry and asynchrony in heart contraction are both critical components of dilated cardiomyopathy that can be improved by CRT. Magnetic resonance asynchrony is efficacious in screening patients and should now be compared with recently published echocardiography data to improve outcome for this costly but valuable therapy. PMID- 16353445 TI - Mechanical testing of human cardiac tissue: some implications for MRI safety. AB - PURPOSE: The effects of aging on tissue strength and its ability to withstand forces associated with MRI have not been investigated. This study aimed to determine the forces required to cause partial or total detachment of a heart valve prosthesis in patients with age-related degenerative diseases exposed to MRI. METHODS: Eighteen tissue samples excised during routine heart valve replacement surgery were subjected to a suture pull-out test using a tensile materials testing machine. Five preconditioning cycles were applied before commencing the final destructive test. The test was complete when the sample ruptured and the suture was pulled completely free from the tissue. Results were compared with previously calculated magnetically induced forces at 4.7 T. RESULTS: All tissue samples displayed a basic failure pattern. Mean forces required to cause initial yield and total rupture were 4.0 N (+/- 3.3 N) and 4.9 N (+/- 3.6 N), respectively. Significant factors determining initial yield were stenosed calcific tissue (p < .01), calcific degeneration (single pathology) (p < .04) and tissue stiffness (p < .01). Calcific degeneration (p < .03) and tissue stiffness (p < .03) were also significant in determining maximum force required to cause total rupture. CONCLUSION: Specific age-related degenerative cardiac diseases stiffen and strengthen tissue resulting in significant forces being required to pull a suture through valve annulus tissue. These forces are significantly greater than magnetically induced < 4.7 T. Therefore, patients with degenerative valvular diseases are unlikely to be at risk of valve dehiscence during exposure to static magnetic field < or = 4.7 T. PMID- 16353446 TI - The case of the disappearing myxoma. AB - We present a case demonstrating the utility of cardiovascular magnetic resonance (CMR) in the diagnosis of a cardiac mass. A 70-year-old female who presented with chest pressure and left sided jaw pain was found to have a cardiac mass on transthoracic and transesophageal echocardiography that was diagnosed as an atrial myxoma. A cardiac magnetic resonance test determined the mass to be more consistent with a thrombus than a myxoma through a stepwise approach using multiple pulse sequences. Thus, unwarranted and potentially risky thoracic surgery was avoided by the incorporation of a systematic evaluation by cardiac MRI. PMID- 16353447 TI - Delayed hyperenhancement in a case of Takotsubo cardiomyopathy. AB - Takotsubo cardiomyopathy (TTC) consists of an acute onset of transient akinesia of various parts of the left ventricle (apex and mid in classical TTC, mid and base in the variant form), without significant coronary artery stenosis, often accompanied by chest pain, dynamic reversible ST-T segment abnormalities and elevation of cardiac enzymes disproportionate to the extent of akinesia. Contrast enhanced cardiovascular magnetic resonance (CMR) is a useful adjunct in the diagnostic work up of patients with TTC. Delayed hyperenhancement on gadolinium enhanced CMR, which is indicative of active inflammation (e.g. myocarditis) or myocardial fibrosis (e.g. myocardial infarction), is usually absent in patients with TTC. In this report we present the case of a 46-years old women with TTC who had an extensive area of apical and midventricular akinesia and in whom gadolinium-enhanced CMR demonstrated a small area of subendocardial delayed hyperenhancement. A gadolinium-enhanced CMR performed 6 weeks later exhibited complete reversal of all wall motion abnormalities and an identical area of subendocardial delayed hyperenhancement. PMID- 16353448 TI - Acute allergic reaction upon first exposure to gadolinium-DTPA: a case report. AB - This is a case report of allergic reaction to Gadolinium-DTPA on first exposure, with a brief review of safety of MR contrast agents. PMID- 16353449 TI - Functional and metabolic recovery of the right ventricle during Bosentan therapy in idiopathic pulmonary arterial hypertension. AB - An impaired high-energy phosphate metabolism might play a critical role in the pathogenesis of right ventricular (RV) failure due to chronic pulmonary arterial hypertension (PAH). 31P-NMR spectroscopy is well established for measurements of high-energy phosphate metabolites in various left ventricular heart diseases, however, mainly for technical and sensitivity reasons, its successful transfer for measurements in the RV is currently missing. In the present study, the usefulness of this non-invasive approach is not only shown in RV failure due to PAH but also tested during subsequent therapy. PMID- 16353450 TI - We must face 2006 together. PMID- 16353451 TI - Now there is a Roth IRA in everyone's future. PMID- 16353452 TI - Gratitude to the late Dr R J Donaldson. PMID- 16353453 TI - Trial by triathlon. PMID- 16353454 TI - Socio-economic status: an insufficiently appreciated component of US health. PMID- 16353455 TI - Bowel cancer: the outcome is improving. AB - The management of rectal cancer has been revolutionised over the last 20 years through improvements in staging, surgery and the use of radiotherapy and combination chemoradiotherapy. The traditional approach involved surgery followed by pathological assessment of the specimen reporting the completeness of excision. Incomplete excision in many cases led to local recurrence of the cancer within the pelvis with the disabling symptoms of pelvic pain, bowel fistulation and bowel and bladder dysfunction. Indeed, many patients died with local recurrence alone and salvage therapy in these patients was rarely successful. However, the recognition of the advantage of pre-operative therapy for patients in whom surgery would not provide local cure has been a major advance. The ability to downstage/downsize the tumour before curative surgery has improved outcome. However, these pre-operative therapies are toxic and not without their own complications, therefore the select use, targeted at the appropriate patient, represents the 'gold-standard'. The development of magnetic resonance imaging staging that accurately predicts operability has completed this triad approach which is focused through the weekly multidisciplinary team meeting. National acceptance of these techniques and standards can only improve the outcome for patients with this disease. PMID- 16353456 TI - HIV infection and circumcision: cutting through the hyperbole. AB - The objective of this study was to determine whether the justifications given for promoting mass circumcision as a preventive measure for HIV infection are reasonable and whether mass circumcision is a feasible preventive measure for HIV infection in developing countries. The medical literature concerning the practice of circumcision in the absence of medical indication was reviewed regarding its impact on HIV infection and related issues. The literature was analysed with careful attention to historical perspective. Our results show that the medical literature supporting mass circumcision for the prevention of HIV infection is inconsistent and based on observation studies. Even if the two ongoing randomised controlled trials in Africa show a protective benefit of circumcision, factors such as the unknown complication rate of the procedure, the permanent injury to the penis, human rights violations and the potential for veiled colonialism need to be taken into account. Based on the best estimates, mass circumcision would not be as cost-effective as other interventions that have been demonstrated to be effective. Even if effective, mass circumcision as a preventive measure for HIV in developed countries is difficult to justify. PMID- 16353457 TI - Knowledge of sexually transmitted infections and sources of information amongst men. AB - The objective of this study was to investigate men's awareness of sexually transmitted infections (STIs) and sexual health services, together with the best ways of providing information about them. Information was collated via a self administered questionnaire, which was answered by 429 men, giving a response rate of 31%. Men aged 16+ living in two primary care trusts (PCTs) in the north-west of England were accessed through a range of different sources, including social, educational, health, work and religious facilities. Despite the high rates of STIs in the north-west, approximately one in five men had not heard of herpes, syphilis or chlamydia. One in ten had not heard of gonorrhoea. The majority of men were unaware that many STIs could be asymptomatic. Only 13% knew where their local genitourinary medicine (GUM) clinic was. The main deterrents to accessing the clinic were embarrassment and not knowing where it was. Men's knowledge of STIs is limited, particularly in relation to the symptoms. The best way of providing information, according to the respondents, was through their general practitioner (GP). However, in practice this might be difficult, as men are reluctant users of GP services in general. Knowledge of the whereabouts of the local GUM clinics is also limited. Although most men thought that they would visit their GP if they thought they might have an STI, this may result from them not knowing the alternatives, rather than having a real choice. More information, targeted wisely, is thus necessary in order to educate men about STIs and service provision. PMID- 16353458 TI - Journalists and gender-based violence in Ibadan, Nigeria. AB - Violence against women (VAW) continues to occur in many countries of the world despite various measures taken to stop it. The media can shape the attitudes and perceptions of people and can be an advocate for social change. Thus an assessment of the knowledge, attitude and coverage of prevention of VAW by 230 media practitioners in Nigeria was conducted using semi-structured self administered questionnaires. Results of the survey revealed that 86 of the 230 (37%) stated that both sexes have equal privileges and rights, while 78 (34%) felt that violence could help to keep women 'in check' or in control. Mean knowledge score out of 10 was 7.5 +/- 2.1, while attitude scores out of 11 were 6.4 +/- 1.2. Two main groups of journalist were identified: prejudiced, (105, 46%), and non-prejudiced (125, 54%). The non-prejudiced group consisted mostly of females (p<0.05), over the age of 36 (p<0.05), and held senior positions within their profession (p>0.05). Female media practitioners had significantly higher knowledge and attitude scores than their male counterparts (7.47 +/- 2.0 versus 6.39 +/- 2.3) and (6.27 +/- 2.0 versus 5.11 +/- 6.1). Ninety respondents (39%) had worked on prevention of VAW in the last two years. Most of these (60, 67%) had worked on one-off programmes. Regular programmes usually consisted of 30 minutes or one-hour weekly programmes on radio and television as well as articles in the newspapers. Enlightening the public (41%) and 'portraying women in a successful light' (12%) were some of the activities the practitioners were ready to embark upon. Media practitioners need first of all comprehensive training on VAW, which should be followed by increased coverage of prevention of VAW in the media. Finally, more interactive media engagement with the public should be employed. Media organisations for women may be useful in initiating some of these changes. PMID- 16353459 TI - Long-term influence of working abroad on returnees' mental health. AB - Although international business travel is increasing, there is a lack of research on its repercussions for mental health. This study analysed the long-term influence of international business travel on the mental health status by comparing depression, anxiety and job stress between workers with and without international assignment experience. The subjects were divided into an 'experienced group' composed of 70 male workers who had experienced an overseas assignment of at least six months, and a 'non-experienced group' consisting of 2,163 male workers who had not. To assess the mental health status, Zung's Self Rating Depression Scale (SDS) and Sheehan's Patient Rated Anxiety Scale (Sheehan) were employed. The Job Content Questionnaire (JCQ) was used to examine job stress. In addition, information about the characteristics of the overseas assignments was collected. The experienced group had significantly higher scores for job control, supervisor support and co-worker support in the JCQ, while no differences were observed for the SDS and Sheehan. Whether or not the subjects travelled abroad with their families, whether or not they went against their will, and whether or not they enjoyed their stay had no effects on their mental health. Job demand had a significantly positive correlation with the duration of the assignment. PMID- 16353460 TI - 15 new developments are already reshaping 2006 Medicare coverage. PMID- 16353461 TI - CMS will limit bariatric surgery benefits to a younger demographic. PMID- 16353462 TI - Two key factors hold down health benefit cost increases. PMID- 16353464 TI - Two decades of hard work. PMID- 16353463 TI - TMJ surgery revisited. PMID- 16353465 TI - Craniofacial morphology and inclination of the posterior slope of the articular eminence in female patients with and without condylar bone change. AB - This study investigated the association of craniofacial and glenoid fossa shapes and temporomandibular joint (TMJ) pathology in 39 orthodontic patients with signs and symptoms of TMJ disorders, using helical CT scans. Cephalometric measurements showed that 21 subjects with bilateral condylar bone change (BBC) had significantly smaller SNB angles, ramus heights and S-Ar/N-Ba ratios, as well as larger mandibular plane angles and lower anterior facial height than the 18 subjects with no condylar bone change (NBC). The average posterior slope of the left and right articular eminence in their central and lateral sections was significantly steeper in NBC than in BBC. Condylar bone change might, therefore, not only be related to the morphology of the mandible, but also of the glenoid fossa and cranial base. This appears to reflect adaptive changes in the condyle, articular eminence and cranial base in response to changes in loading. PMID- 16353466 TI - Effect of interocclusal appliance on masticatory performance of patients with bruxism. AB - This study was designed to evaluate masticatory performance in bruxist patients. The experimental group was composed of 16 dentate volunteers presenting with bruxism and treated by interocclusal appliances and a control group, consisting of 16 dentate volunteers with no symptoms of bruxism. To evaluate masticatory performance, the volunteers chewed .03 grams of peanuts contained in polyvinyl wrapper for four and twelve masticatory cycles. The number of chewed particles was counted using an optical scanning method by Image-Pro Plus 1.4 software (Media Cybernetics, Inc., Silverspring, MD). The score was expressed as the mean of the number of chewed particles for each wrapper. Masticatory performance evaluations were carried out in a single period for the control group and for the experimental group, before interocclusal appliance insertion, and after seven, 15, 30, and 60 days. Comparison between groups, as well as between before and after interocclusal appliance insertion, showed no significant differences (P>.05), irrespective of the number of masticatory cycles. PMID- 16353467 TI - Evaluation of body posture in individuals with internal temporomandibular joint derangement. AB - Temporomandibular dysfunctions (TMD) comprise a great number of disruptions that may affect the temporomandibular joint (TMJ), the masticatory muscles, or both. TMJ internal derangement is a specific type of TMD, of which the etiology and physiopathology are broadly unknown, but have been suggested to be linked to head, neck, and body posture factors. This study aimed at verifying possible relationships between body posture and TMJ internal derangements (TMJ-id), by comparing 30 subjects presenting typical TMJ-id signs to 20 healthy subjects. Subjects' clinical evaluations included anamnesis, stomatognatic system evaluation, and plotting analysis on body posture photographs. No statistically significant differences were found between the groups. Results do not support the assertion that body posture plays a role in causing or enhancing TMD; however, these results should be cautiously considered because of the small number of subjects evaluated and the many posture variables submitted to statistical procedures that lead to high standard deviations. PMID- 16353468 TI - Effect of short-term use of a centric occlusion stabilization oral appliance on sensory and pain perception thresholds in the cervically innervated area: a pilot study. AB - This preliminary study was done to determine the effect of short-term use of a centric occlusion stabilization oral appliance with regard to noxious and sensory perception in the upper extremities. The subjects consisted of 22 asymptomatic females, and the experiments were performed on two separate days within the same week, with the days randomly assigned as either appliance-wearing or nonappliance wearing days. For each experimental day, cool sensation, warm sensation, cold induced pain and heat-induced pain were measured using a computer-based quantitative testing device, and these thresholds were compared between the experimental days with or without the oral appliance. We found that during the experimental day wearing an oral appliance, subjects had significantly higher thresholds for warm sensation and heat-induced pain. There was no statistical difference between the testing days in cool sensation or cold-induced pain thresholds. These findings indicate that short-term wearing of a centric occlusion stabilization oral appliance may inhibit some noxious and sensory inputs from cervically innervated structures, which are primarily served by unmyelinated C fibers, and that oral appliances may be appropriate for the treatment of painful cervical disorders. PMID- 16353469 TI - Prolotherapy (proliferation therapy) in the treatment of TMD. AB - Proliferation therapy, or "prolotherapy," is also known as regenerative injection therapy (RIT). Since the 1930s, the technique has been used to stabilize injured joints and to relieve joint pain. This article reviews the history and scientific literature regarding prolotherapy and describes the application of the technique to treat injured or unstable temporomandibular joints (TMJ). Alternative medicaments and the likely mechanisms of action are discussed. A brief preliminary summary of a retrospective clinical study of the efficacy of prolotherapy is included. The study shows that prolotherapy can be an effective therapeutic modality that reduces TMJ pain and joint noise in a majority of patients who have reached a plateau with use of an intraoral appliance, physical therapy, and home care. PMID- 16353470 TI - Full-mouth rehabilitation following treatment of temporomandibular disorders and teeth-related signs and symptoms. AB - The literature is replete with theories regarding temporomandibular disorders (TMD). However, there is a paucity of information concerning perceived malocclusion and other teeth-related signs and symptoms after full-mouth rehabilitation. This clinical study was designed to evaluate the perception of TMD patients concerning perceived malocclusion and other teeth-related signs and symptoms after full-mouth rehabilitation guided by the Mental Analog Scale (MAS). Among 38 patients referred for full-mouth rehabilitation, 20 were diagnosed as having TMD after reviewing a questionnaire, recording the major complaints and symptoms, in addition to performing comprehensive clinical examination. Nonsurgical therapy was performed, including fabricating an anterior programming device, a centric relation occlusal device and finally full-mouth rehabilitation by means of placing crowns on all upper and/or lower teeth. All full-mouth rehabilitation procedures were performed using a fully adjustable articulator and mandibular movements were recorded following pantographic tracings. After full mouth rehabilitation, the patients were followed up at 1, 2, 4, 6, 9, and 12 month intervals, and the major signs and symptoms were recorded along with adjunctive teeth-related signs and symptoms. Fisher exact probability tests were applied to analyze the results (P<.05). Statistical comparisons of the MAS responses before and after treatment (at 1-month recall) showed significant improvement (P<.05) for all teeth-related signs and symptoms except for bruxism (P=.0699). Further improvement was noted at the 4-month recall period. However, these improvements were not statistically significant for all teeth-related signs and symptoms. No further change was noted after the 4-month recall period. There was a marked reduction in perceived malocclusion and adjunctive teeth-related signs and symptoms during function, only after performing occlusal equilibration of the final restorations. PMID- 16353471 TI - Trifid mandibular condyle: a case report. AB - A suspected case of trifid mandibular condyle that was observed on a panoramic radiograph and confirmed using computed tomography (CT) is reported. No history of previous temporomandibular joint (TMJ) trauma was reported by the subject. PMID- 16353472 TI - Guidelines for anterior repositioning appliance therapy for the management of craniofacial pain and TMD. PMID- 16353473 TI - [Sensitivity of cells to oxygen and erythropoietin production]. AB - The review describes physiological responses of cells to oxygen concentration mediated mostly by action of hypoxia-inducible factor-1, as well as the role of this factor in erythropoietin production and induction of mammalian genes by hypoxia. PMID- 16353474 TI - [Reproductive function and viability of progeny in female rats under unfavourable influences in run up to puberty]. AB - Wistar female rats were subjected to a 3.5-day water deprivation once a week in the period of 1.5 to 3 months of age. Their progeny was subjected to the same influences during the same period of life. A week later, reproductive function of female rats was evaluated by mating them with normal males. In the experimental groups of both generations, no significant changes were found in number of neonates and their body mass, in maternal behaviour during the lactation period, in postnatal mortality of pups, in their growth and development, in motor activity, physical endurance and behaviour. PMID- 16353475 TI - [Features of a passive avoidance extinction of mice with a different level of anxiety]. AB - The dependence of the passive avoidance extinction from a level of mice initial anxiety is investigated. Mice were classified as high-, medium- and low-anxious by time spent in the open area of the elevated plus-maze. It is revealed that to each from levels of anxiety there corresponds certain dynamics of extinction. The high-anxious mice are characterized absence of the passive avoidance extinction and stability of good retrieval of memory trace for want of testing down to 15 days. At medium-anxious mice the deficit of avoidance habit performance developed since the 7th day of extinction. At low-anxious mice since the 11th day of testing the deterioration of retrieval was observed. PMID- 16353476 TI - [The behaviour and respiration features in rabbits with different pattern of their open field locomotor activity]. AB - Behavior of 19 Chinchilla rabbits was tested in the open field. By the results of the first trial, animals were divided into three groups by the type of the exploratory locomotor activity (passive--37%; active--21; and medium--42%). During the repeated open field trials, the passive rabbits increased their activity and the active ones decreased it. Correlations were found between the open field behavior and patterns of the external respiration in the absence of movements. The longest respiratory cycles, expirations and respiratory breath holdings were observed in passive rabbits. Animals with prevailing right turnings in the open field displayed longer respiratory cycles and expirations than those with prevailing left turnings. It seems that the pattern of external respiration can predict the passive defensive strategy of rabbits in negative emotional situations. PMID- 16353477 TI - [Analysis of heat transfer in the biological tissue layer with distributed system of blood vessels]. AB - Processes of heat transfer in the skin layer with blood vessels were investigated using mathematical modeling. Analysis of influence of a pathological state of blood vessels on heterogeneity of thermal field of the skin surface was carried out. For each site of body surface, there is a certain difference of temperature between normal and pathological sites, being specific for differential diagnosis of diseases of dermal and hypodermic vessels. PMID- 16353478 TI - [Possible mechanisms of unilateral hippocampal insult after bilateral occlusion of common carotid arteries in rats with different types of behavior]. AB - Possible mechanisms of unilateral ischemic insults in CA1 region of hippocampus and anterior dorsal nuclei of thalamus after bilateral occlusion of common carotid arteries in rats with different types of behavior are discussed. PMID- 16353479 TI - [Blood oxygen transport in rats during lipopolysaccharide administration combined with modification of L-arginine-No system]. AB - Lipopolysacharide from Escherichia coli was intravenously administered to rats (5.0 mg/kg). L-arginine-No system was modified by intravenous injection of L arginine, N(G)-nitro-L-arginine methyl ester or L-lysine-N(G)-acetamidine (nitric oxide synthase (NOS) substrate, nonselective NOS inhibitor, and selective inducible NOS inhibitor, respectively.) Lipopolysacharide-induced disorders of blood oxygen transport were the least during the selective inducible NOS inhibition. The protective effects of L-arginine and N(G)-nitro-L-arginine methyl ester were less prominent. Such features of NOS modification effect on the blood oxygen transport suggest that activation of inducible NOS may change the hemoglobin-oxygen affinity during the lipopolysacharide treatment. PMID- 16353480 TI - [Effect of corticoliberin fragment CRF(4-6) on pain sensitivity in rats]. AB - The effects of corticoliberin fragment CRF(4-6) (Pro-Pro-Ile) on pain sensitivity of rats in "hot plate" test were investigated. Intracerebroventricular administration of tripeptide CRF(4-6) (6, 30, 150 nmol/head) induced dose dependent antinociception: the latency of paw lick response increased by 7.4 +/- 1.4, 10.1 +/- 1.5 and 16.7 +/- 4.2 s respectively from the basic level of 10.2 +/ 0.9 s. Duration of tripeptide antinociceptive action was 30 min (for 6 nmol) and 60 min (for 30 and 150 nmol). Pretreatment with corticotropin-releasing factor antagonist alpha-helical CRF(9-41) (6.5 nmol/head) 60 minutes before tripeptide administration completely abolished the antinociceptive effects of CRF(4-6) (6 nmol). Therefore corticoliberin receptors seem to be involved in realization of tripeptide influence on pain sensitivity. The data obtained suggest that CRF(4-6) can either directly interact with corticoliberin receptors or modulate activity of CRF-ergic neurons. PMID- 16353481 TI - [Age- and sex-related differences in sensitivity to hepatotoxic action of estragole in mice]. AB - As determined by blood activity of alanine- and aspartate aminotransferases, intraperitoneal injection of estragole in subcarcinogenic dose 300 mg/kg does not damage the liver of suckling off-spring of both sexes and of adult SWR/J males but drastically damages it in mature females of this strain. Castration only slightly decreases the resistance of males to hepatotoxic action of estragole but significantly increases it in females; exogenous administration of estradiole benzoate to castrated males decreases their resistance to the hepatotoxin, whereas administration of testosterone propionate to ovariectomyzed females does not elevate it. Morphologically, estragole damages the same number of liver cells in females and males, but in males it induces mostly hydropic degenerative, and in females--necrotic changes of hepatocytes. PMID- 16353482 TI - [Behavioral activity and stress reaction of the pituitary-adrenocortical axis in rat with prenatal inhibition of testosterone metabolism]. AB - The effects of maternal administration of the aromatase inhibitor, 1,4,6 androstatrien-3,17-dione (ATD), during the last week of gestation on stress reaction of the hypothalamic-pituitary-adrenal axis (HPA) and behavior in a novel environment (open field) and the anxiety level in the elevated plusmaze, were studied in male and female adult offspring. The results showed that parental inhibition of brain testosterone metabolism decreases the basic level of corticosterone in male rats and prolongs hormonal stress reaction of the HPA axis in both sexes. Prenatally treated rats demonstrated significant elevation of the anxiety level and emotionality. There was no sexual dimorphism in behavioral response to a novel environment such as locomotor activity, the time of immobilization, the total duration of grooming reaction, and the anxiety level, between control male and treated female rats. These data suggest a prenatal inhibition of the brain testosterone metabolism after the stress reaction of HPA axis and formation of sexual dimorphism in the anxiety and behavioral response to a novel environment in adulthood. PMID- 16353483 TI - [Visual mechanisms of recognition of vanishing figures]. AB - The acuity test Landolt C's with a complex contour imitating various kinds of high-pass spatial frequency filtering generated as a printed visual acuity charts with a grey background were studied with regard to recognition thresholds. Comparisons were made for recognition thresholds with different complex contours. When the vanishing optotypes are out of focus, their image on the retina fades rapidly into the grey background, rendering them invisible rather than merely blurred as in the standard chart. The data obtained confirm that the recognition distance depends on physical property of complex contour profile and show the importance of the Landolt C spatial frequency spectrum for recognition of different types of the complex contour. PMID- 16353484 TI - [The dynamic of breathing biomechanics and intrathoracic hemodynamics parameters during microgravity modeling]. AB - In ground-based model of the hemodynamics effects of weightlessness, the intersystem relation of breathing and circulation was investigated during inspiration and expiration separately in anesthetized catz. It's shown that the dynamics of central venous pressure, esophageal pressure and filling pressure of the heart during inspiration in supine and head-down tilt position has obvious similarity to those which hypothetically can be present in microgravity. The results suggest that intrathoracic hemodynamics during inspiration in supine and head-down position may be an adequate ground model for investigation of weightlessness influences on intrathoracic circulation. PMID- 16353485 TI - [Coupling of protease activity and sodium loading with intestinal absorption of amino acids]. AB - Membrane-bound serine proteases to play a certain role in activation of sodium transport in epithelial cells. To were found explain the protease activity and sodium-dependent L-tryptophan transport across chicken small intestine interaction, four experiments were conducted. One hundred chicks were fed diets that contained 0; 0.3; 3 or 6% of supplemental NaCl and were given distillated water ad libitum. Signs of salt toxicity observed were as follows: a decreased body weight, increased heart and kidney weights, formation of secondary lysosomes in enterocytes and lymphocytes. Such chickens were in the state of negative nitrogen balance. Intestinal absorption of L-tryptophan correlated with mucosal protease activity during increased dietary sodium chloride intake. Recent in vitro and in vivo experiments indicate that enterocyte proteases may be of critical importance in activation of sodium-dependent intestinal transporters for L-tryptophan. PMID- 16353486 TI - [Modification of electrode for regional mucose blood flow measurements with the aid of hydrogen gas clearance]. PMID- 16353487 TI - LYVE-1 expression on high endothelial venules (HEVs) of lymph nodes. AB - LYVE-1 (lymphatic endothelium hyaluronan receptor) has been identified as a powerful marker for lymphatic endothelium. Apart from lymphatic endothelium, LYVE 1 is expressed in normal liver blood sinusoids, spleen endothelium and activated tissue macrophages. LYVE-1 has not been detected in blood vascular endothelium with the exception of blood vessels in the lung. High endothelial venules (HEVs) belong to the vascular compartment of lymph nodes. They are the major site of entry for circulating lymphocytes into the node. HEVs are characterized by cuboidal endothelial cells, the existence of discontinuous junctions between these endothelial cells, and the presence of large numbers of lymphocytes within their walls. 40 paraffin-embedded lymph node biopsy specimens from newly diagnosed patients with non-Hodgkin lymphoma were evaluated as well as 10 lymph node biopsy specimens from adult patients with reactive lymphadenitis, and 10 normal, non-metastatic lymph nodes obtained from adult patients during cancer surgery served as controls. Samples were fixed in 10% buffered formalin, paraffin embedded, and stained with hematoxylin and eosin for histopathological evaluation. Sections were also evaluated with mouse monoclonal antibodies against LYVE-1 and CD34, and expression of both LYVE-1 and CD34 was demonstrated in HEVs. LYVE-1 expression was also found on the endothelial cells of the lymphatic sinus and in reticular cells in the lymph nodes. PMID- 16353488 TI - Pulmonary lymphangiectasia. AB - Congenital pulmonary lymphangiectasia (PL) is a rare developmental disorder involving the lung and is characterized by pulmonary subpleural, interlobar, perivascular, and peribronchial lymphatic dilatation. Both frequency and etiology are unknown. PL presents at birth with severe respiratory distress, tachypnea, and cyanosis, with a very high mortality rate at or within a few hours of birth. At birth, mechanical ventilation and pleural drainage are nearly always necessary to obtain a favorable outcome of respiratory distress. Home supplemental oxygen therapy and symptomatic treatment of recurrent cough and wheeze are often necessary during childhood, sometimes associated to prolonged pleural drainage. Recent advances in intensive neonatal care have changed the previously nearly fatal outcome of PL at birth. Patients affected by PL who survive infancy present medical problems which are characteristic of chronic lung disease. PMID- 16353489 TI - New clinical and laboratory staging systems to improve management of chronic lymphedema. AB - We have developed new clinical (C) and laboratory (L) staging systems to improve the clinical management of chronic lymphedema. These systems were retrospectively assessed in 220 chronic lymphedema patients followed up for 4 years. Clinical evaluation of the treatment response/disease progression was performed at 6 month intervals and laboratory evaluation at a yearly interval except for recurrent sepsis cases. The reliability of C-stage and L-stage for the progression of disease were analyzed separately. The C-staging was based on the subjective and objective findings of local and systemic conditions, while L-staging was based on lymphoscintigraphicfindings. Clinical implementation of this new staging system facilitated interpretation of the progress/deterioration of the clinical response to CDT treatment, and it was found to be a useful guideline for the decision/selection of further surgical treatment. We propose that these two separate staging systems could now become a new guideline for improved management of lymphedema with a better prediction of treatment outcome and decision point for additional medical/surgical therapy. Further clinical implementation and evaluation is necessary to demonstrate clinical usefulness especially to guide surgical therapy and L-staging in followup. PMID- 16353490 TI - Differential expression of collagens type I and type IV in lymphangiogenesis during the angiogenic process associated with bleomycin-induced pulmonary fibrosis in rat. AB - In order to assess the role of collagens I and IV during the angiogenic process associated with bleomycin-induced pulmonary fibrosis in rat, in situ hybridization and immunocytochemical studies were carried out. An increased expression of collagen IV was observed before an enhanced expression of collagen I after intratracheal instillation of bleomycin. Deposits of both collagen types were detected on the 21st day after treatment with bleomycin, surrounding the new blood vessels formed during the fibrotic process. At this time, the presence of new lymphatic vessels was associated uniquely with deposition of collagen I. These observations lead us to conclude that, at least during pulmonary fibrosis, lymphangiogenesis takes place after blood angiogenesis. PMID- 16353491 TI - The effect of gentle arm exercise and deep breathing on secondary arm lymphedema. AB - The aim of this study was to explore the benefits of gentle arm exercise combined with deep breathing for secondary arm lymphedema. 38 women participated in 10 minutes of standardized arm exercise and deep breathing and were measured every 10 minutes for 1 hour, then 24 hours and 1 week post regime. A smaller cohort of 24 women continued the 10 minute exercise regime morning and evening for 1 month, with measurements being repeated at the end of this time. Directly after performing the regime, there was a reduction in arm volume of 52 mls (5.8%), with the reduction being sustained at 30 minutes (50 mls, 5.3%). Even though participants were told not to further do the exercise, at 24 hours the volume reduction was 46 mls (4.3%) and at 1 week, 33 mls (3.5%). At the one month follow up, the reduction was 101 mls (9.0%). All reductions were statistically significant. Reported arm heaviness and tightness also statistically significantly decreased directly after the regime with the reduction in tightness being sustained at 24 hours. The reduction in heaviness was sustained at 24 hours, 1 week, and even one month after the program. Perceived limb size was significantly reduced at 1 week and at the 1 month follow-up. There was also a significant improvement in the anterior thorax tonometry reading at the 1 month follow-up. PMID- 16353492 TI - Influence of angiostatin and thalidomide on lymphangiogenesis. AB - Malignant cancers commonly invade locally followed by spread through venous or lymphatic channels or both to distant sites. Hemangiogenesis and its relation to tumor growth and metastasis have been extensively studied. However, the role played by lymphangiogenesis in growth and metastasis of cancer has been largely neglected until just recently. Inhibition of lymphangiogenesis, as compared to inhibition of hemangiogenesis, may provide new insights into the mechanisms of cancer metastasis. The current study was designed to examine the in vitro effect of two commonly used inhibitors of hemangiogenesis, angiostatin and thalidomide, on the growth and proliferation of lymphatic endothelial cells isolated from pig thoracic ducts. We first isolated and characterized the lymphatic endothelial (LE) cells using specific markers for VEGFR3 and LYVE-1. The experimental results showed that treatment of the LE cells with these two drugs resulted in a decrease in the rate of cell proliferation in a dose-dependent manner as assessed by MTT assays. Cell migration rate was assessed by the speed of cell migration from the scrape-wound margin, and the results showed that migration of LE cells was also significantly inhibited in a dose-dependent fashion compared to controls. Treatment with angiostatin and thalidomide both resulted in an increase in apoptosis of LE cells as assessed by Hoechst staining and flow cytometry. We conclude that both angiostatin and thalidomide are able to inhibit LE cell growth in a dose-dependent manner and that the inhibition may be through induction of apoptosis. PMID- 16353493 TI - Effective error reporting: quality leaders share cutting-edge strategies. PMID- 16353494 TI - Measuring the value of patient education. PMID- 16353495 TI - Patient satisfaction planner. Program improves care, hospital collaborations. PMID- 16353496 TI - Patient satisfaction planner. Community interventions aid self-care, prevention. PMID- 16353497 TI - Maryland ICUs collaborate to revamp care processes. PMID- 16353498 TI - Study: public's interest in quality data is growing. PMID- 16353499 TI - Bird flu pandemic may not be certainty, but it's not too early to start preparing. AB - Revisiting your policies and procedures and intensifying your preventive practices will not only enhance the health of your staff, but it also will improve patient safety. Urge all of your staff to have a flu vaccination this winter. Encourage the use of protective gear, such as gloves and masks, when treating patients with coughs or colds. Have staff who are ill stay at home, rather than risk exposure of colleagues and patients. PMID- 16353500 TI - Some chest pain patients may be discharged early. AB - Your clinicians may wish to consider this new rule when treating chest pain patients, and then make their own determination as to treatment. It should not be incorporated into a clinical pathway. Patients younger than 40, with normal initial EKG and no prior history of ischemic chest pain, may be suitable for early discharge. Older patients also may be eligible for early release, if cardiac enzymes are low or do not change or rise in two hours. If your ED does not have a chest pain observation unit, you may end up admitting a greater percentage of these patients. PMID- 16353501 TI - 'Virtual beds' lower flow times, boost satisfaction. AB - Here's an approach that makes optimal use of space and time in the ED. It's not as easy as it sounds, however; be prepared to work hard at communications between your nurses and physicians. Take patients who potentially can be discharged quickly, put them in a room, examine them, then discharge them or put them back in a waiting room after ancillary testing. Order lab or radiology testing up front. Patients may be taken to those departments or testing is done in another area of the ED. Identifying nonurgent patients is a useful strategy for overcoming nurses' objections. One ED tried the system, but abandoned it due to volume increase. PMID- 16353502 TI - Katrina-born tracking forms aid Rita response. AB - Major disasters can lead to thousands of patients being evacuated. It's essential to track their whereabouts in a single source that is readily available to other facilities as well as concerned relatives. Centralized form should include most critical information, including patient's name, date of birth, and chief complaint, as well as hometown and how they were transported. Make this information available to local authorities and the American Red Cross. Trade patient information with other health care facilities. Expand the reach of your database as far geographically as possible. Patients may be moved to other cities throughout your state. PMID- 16353503 TI - Neuroplasticity, learning and recovery after stroke: a critical evaluation of constraint-induced therapy. AB - Constraint-induced movement therapy (CIMT) has been hailed as a radical new approach to stroke rehabilitation. The guiding theory is that impairment of hand function is exacerbated by learned non-use and that this in turn leads to a loss of cortical representation of the upper limb. It is claimed that these processes can be reversed by two weeks of constraint of the unaffected limb combined with intensive practice in use of the paretic hand, and numerous small-scale studies have suggested that CIMT can lead to large improvements in function more than a year after stroke. However, the theory of learned non-use is open to question and there is uncertainty about the nature of the improvements induced by CIMT. The greatest effect seems to be increased spontaneous use of the hand, either through reduction of learned non-use or by overcoming the sense of effort during movement. There is also evidence of some improvement on dexterity tests but no studies have analysed in detail whether this reflects reduction of basic motor impairment or learning of compensatory movement strategies. The current weight of evidence is in favour of compensatory learning. Cortical changes detected by transcranial magnetic stimulation (TMS) or functional imaging may reflect this compensatory motor skill learning rather than restoration of representations lost due to the infarct or non-use of the limb. If future studies confirm this then the clinical implication is that direct teaching of unimanual or bimanual compensatory strategies might be a more productive approach than constraint. PMID- 16353504 TI - Developing functional outcome measures for unilateral neglect: a pilot study. AB - Stroke patients may develop personal neglect, peripersonal neglect or both. Four new measures were tested in a sample of 42 right-handed inpatients (25 male; 17 female, median age 72 years). Participants removed keys from a rack, identified grocery items, washed their face, and cleaned a tray. Prior to this, they were classified as: no neglect (15), personal neglect (8), peripersonal neglect (7), and both personal and peripersonal neglect (12). The sensitivity and specificity of each new measure was determined by agreement with the classification. Test retest reliability was determined using weighted kappa statistics or limits of agreement. Four occupational therapists (OTs) rated videos of the face and tray measures, and software was developed to measure objectively time spent and area covered on Face and Tray. Keys and Grocery had high specificity, good reliability but poor sensitivity. For the OTs' video ratings, there was good and moderate inter-rater reliability on Tray and Face respectively for area covered, but not time spent. Intra-rater reliability was also better for area than time on Tray. However, the validity of Face and Tray themselves is currently inadequate. A longitudinal study is proposed to modify the measures, increase their sensitivity and evaluate their ability to monitor change over time. PMID- 16353505 TI - A single case experimental design comparing two memory notebook formats for a man with memory problems caused by traumatic brain injury. AB - There is extensive literature demonstrating that restoration of memory is not a realistic goal for rehabilitation and that efforts are more effective if focused on developing compensatory strategies. This paper compares two formats of memory notebook using an ABAB single-case experimental design with a 46-year-old man with a history of head injury. Results revealed significantly better performance on a series of prospective memory tasks for the "modified" notebook. The study highlights the importance of tailoring the diary to the patient's needs and giving careful consideration to the way in which the diary is introduced. PMID- 16353506 TI - Acute cognitive and neurobehavioural intervention for individuals with acquired brain injury: preliminary outcome data. AB - The present study provides a rationale for and detailed description of a structured curriculum for a cognitive and neurobehavioural group intervention for patients in an acute inpatient brain injury rehabilitation setting. Preliminary outcome data are provided for 29 patients with acquired brain injuries who attended the group during inpatient rehabilitation. The group was held during three 30-minute sessions per week. Prior to discharge, patients completed a Learning Assessment, which assessed their level of knowledge about the material covered during the course. Patients received a mean score of 85.54 on the Learning Assessment. High scores on the Learning Assessment correlated significantly with discharge Disability Rating Scale total and Social Interaction FIM scores. Age, level of education, race, sex, and length of stay did not significantly impact these results. Results suggest that patients with acquired brain injuries benefit from acute cognitive and neurobehavioural intervention and are capable of learning compensatory strategies, even in the acute stages of recovery. This learning may help improve functional status, especially skills needed for psychosocial adjustment. PMID- 16353508 TI - [Vaccination: choosing between individual desire and public policy]. PMID- 16353507 TI - Neuropsychological rehabilitation in a case of Cornelia de Lange syndrome. AB - Cornelia de Lange syndrome is a rare congenital disorder. Symptoms include a characteristic facial structure, pre- and post-natal growth deficiency, hypertrichosis, and visceral and cardiovascular anomalies. Behavioural problems and moderate to severe mental retardation are also present. In this paper, we report a mild case of Cornelia de Lange syndrome. The neuropsychological rehabilitation of this patient is also described. It was structured on the patient's specific areas of strength and weakness in order to facilitate the development of adaptive cognitive abilities. To maximise the learning potential of these children, we recommend specific interventions with a multidimensional neuropsychological approach, which considers developmental abilities and disabilities as related to the global reality of the child. PMID- 16353509 TI - [Compliance and resistance to child vaccination: a study among Swiss mothers]. AB - BACKGROUND: This study examines mothers' practices and attitudes in relation to their child's immunization. In this area, like in others, gaps are being observed between the public health model of risk management and the population expectations and behaviors. METHODS: Data were collected using a standardized questionnaire from 1295 women having, in 1999, one child between 7 and 8 years of age of Swiss nationality and attending the public school system in Geneva. RESULTS: Four immunization types were established based on (1) the degree of satisfaction (in relation to past choices), (2) the perception of adequate knowledge about immunization and (3) practices. The mothers were divided into the following categories: compliant (57%), compliant ambivalent (19%), moderately resistant (17%) and resistant (7%). Mothers having reached an intermediate education level are more likely to be resistant. Furthermore, women having negative attitudes towards biomedical institutions, having consulted alternative practitioners and those considering that they have a certain individual control over the health of the family are more often resistant to immunization. CONCLUSION: Our results confirm the existence of a resistance to child immunization among a section of the population. It should not be attributed to mothers' ignorance, but rather could reflect their perplexity towards the choices they are expected to make. In that regards, the importance some parents give to individual control over health can be in contradiction with community objectives of public health. Actions for the promotion of immunization should primarily target families who are uncertain (compliant ambivalent and moderately resistant). At the same time, the difficulties entailed in the - individual and collective - management of risks require that new forms of dialogue be developed between experts and the public. PMID- 16353510 TI - [Periconceptional folates and the prevention of orofacial clefts: role of dietary intakes in France]. AB - BACKGROUND: Orofacial clefts are among the most frequent congenital malformations at birth with a prevalence of 1 in 700 births in Europe. The implication of environmental factors in their etiology has been demonstrated. The role of folic acid, or folates, in the prevention of orofacial clefts is still debated although its efficacy has been demonstrated for the prevention of neural tube defects. METHODS: A case-control study was conducted in 7 hospitals in 4 centers in France. Cases (N=240) were children with non-syndromic orofacial cleft referred to one of the study hospitals for initial surgical repair in 1998 and 1999. Controls (N=236) matched for gender, geographic origin, and age were chosen in paediatric departments of the same hospitals. Usual dietary intake of folates was estimated using a food frequency questionnaire submitted to the mother at hospital. During the same interview, data on sociodemographic characteristics, medical and obstetrical history, tobacco and alcohol consumption, and vitamin supplements, were obtained. Odds ratios associated with quintiles of dietary intake of folates were estimated using logistic regression adjusting for known confounders. RESULTS: A significant dose-response relationship between the risk of orofacial clefts and a decrease in the intake of folates from diet was found, stronger for cleft palate without cleft lip. Only few women (<1%) declared having taken vitamin supplements containing folic acid when planning their pregnancy. CONCLUSION: Our study further suggests that folates are useful for the prevention of orofacial clefts during the periconceptual period. In our study, the estimated average daily intake of folates (270 microg/day) was below French national recommendations for the prevention of neural tube defects (400 microg/day). Since those insufficient folate intakes during the periconceptual period are not presently met by vitamin supplementation, the solution may come from the food fortification program proposed and implemented by the National Agency for Food Safety (AFSSA). PMID- 16353511 TI - Perinatal care regionalization and acceptability by professionals in France. AB - BACKGROUND: For twenty years, most of industrial countries developed recommendations on regionalization of perinatal care. Perinatal regionalization is particularly aimed at improving morbidity and mortality outcomes of low birth weight newborns by transferring pregnant women to the maternity units having a medical or neonatal environment suited to the risks incurred by mothers or babies. Perinatal regionalization cannot be effective without being well accepted by the majority of professionals. The objectives of this study were then to identify professionals'expectations and objections to perinatal regionalisation and to compare them from a professional group to another one. METHODS: Professionals of 3 French perinatal networks were under consideration: the Rhone, the Auvergne and the Gard-Lozere networks. The study included two stages: 1) a psychosociological qualitative study, based on professionals'interviews, aimed at identifying main concerns of professionals and developing a questionnaire; then 2) an epidemiological quantitative study, using this questionnaire within French networks. In the questionnaire, 8 dimensions explored the professionals'views: constraints related to regulation aspects and to the setting up of maternity units care levels, risk of loss of professionals' competence and prestige, consequences on medical practices, on inter-professional relationship, on work organization and financial aspects, and related to the new role of 'private practice'professionals, legal consequences. RESULTS: The response rate of the epidemiological study was 80%. The results permitted to construct 8 dimension scores describing the reasons of poor acceptability of regionalization. After taking into account the age, the sex, the network and the juridical status of the institution, the study revealed a significant poorer acceptability of regionalization by most of medical specialty groups (anesthetists, obstetricians, midwives and "private practice" professionals) compared with neonatologists, or by "private" professionals (professionals working in private clinics and "private practice" professionals) compared with professionals working in university or community hospitals. The study described also network setting up conditions related to its functioning. CONCLUSION: By identifying clearly professionals 'objections and expectations, this study should facilitate improvement in the organization of studied perinatal networks. PMID- 16353512 TI - [Early postpartum hospital discharge in France]. AB - BACKGROUND: Early postpartum discharge is a recent practice in France and for which there are few national data. METHODS: The Sentinel AUDIPOG network was used to describe the practice of postpartum early discharge (< 3 days in case of vaginal delivery and < 5 days in case of caesarean section) from 1994 till 2002, after standardization on the distribution according to the type of hospital and the region (n = 128232). Secondarily, the prognostic medical factors of an early postpartum hospital discharge were researched, in univariate analysis and logistic regression analysis, of the births of 2001-2002. RESULTS: Early postpartum discharges concerned 3% of the deliveries in 1997 and 7% in 2002. Eearly postpartum discharge was more frequent in the level II and III obstetric facilities, in non- university hospitals, in facilities with more than 1 500 deliveries a year, in urban hospitals and in Paris and the surrounding area. The women leaving early in the postpartum were more often multiparas, with no pregnancy pathology, with a single pregnancy, without postpartum hemorrhage, and a child > 2500 g without risk of infection. At present, 40% of vaginal delivery and 25% of caesarean section primiparas and 55% of vaginal delivery and 30% of caesarean section multiparas could be discharged early. CONCLUSION: The reduction of the postpartum hospital stay is inevitable but it is advisable to take care that the women who are discharged are medically fit for discharge. PMID- 16353513 TI - [Socioeconomical status, health behaviors and health status of French adolescents: Health Behavior in School-aged Children (HBSC) survey]. AB - BACKGROUND: The impact of living conditions on health is not well known, but health inequalities observed in adults seem partly determined by behaviours and health status at an earlier stage, and more particularly during adolescence. So, our aim was to study adolescents' health and their health behaviours function to family socioeconomic status. METHODS: We analysed French data from the international survey "Health Behavior in School-aged Children" carried out in a representative sample of adolescents aged 13 and 15 years in 1998. A self administered questionnaire was completed by the adolescents who answered questions concerning their health, health behaviours, and their families' socio economic status. RESULTS: Adolescents from blue-collar families were more numerous to report to be in "fairly good health" or "not very good health" (OR = 1.40, CI 95% = 1.12-1.74) and to be overweight or obese (OR = 1.85, CI 95% = 1.25 2.24) than those from executive families. They were also more numerous to have bad health behaviours than those from executive families. CONCLUSION: This study has established that, in adolescents, differences in health, health behaviours, types of consumption and physical activities depended on the families' socio economic status. PMID- 16353514 TI - [Changes in biological test practices after publication of national guidelines for lipid management among patients under statin treatment]. AB - BACKGROUND: Guidelines for screening and treatment of dyslipidemia were disseminated in September and October 2000 by the National Agency of Accreditation and of Evaluation in Health (ANAES) and the French Agency of Medical Safety of the Products of Health (AFSSAPS). It was confirmed that the specific biological test was the measurement of LDL Cholesterol. OBJECTIVE: To study changes in biological test practices after diffusion of guidelines among patients on statin therapy, using Health Insurance database on reimbursement of patients living in Ile-de-France region (8,534,623 social insurance contributors). METHODS: Two groups of patients were defined in the database from the codes for medication refunds during one month (March 2000 and March 2002). The first group named "new users" included patients starting statin therapy in March, in order to follow the biological test for screening. The second group named "long term users" included patients who had been treated by statin therapy for one year or more, in order to examine the biological follow-up of treatment. All lipid biological tests were recorded during one year, before March 2000 and March 2002. Changes in medical practices were noted as the percentage of the biological tests for "exploration of a lipidic anomaly" (EAL) with determination of LDL cholesterol. RESULTS: For new users the percentage of patients having had at least one EAL for screening purposes increased by 13.5 (39.9% in 2000 and 53.4% en 2002). For long term users the change was + 21.1 (38.3% in 2000 and 59.4% in 2002) during follow-up. CONCLUSION: An improvement in biological testing practices was noted after diffusion of guidelines. PMID- 16353515 TI - [Prevention of infective endocarditis: are recommendations about oral health procedures for high or moderate risk patients respected?]. AB - BACKGROUND: Infective endocarditis (IE) is a rare but particularly serious disease. It frequently requires surgical treatment with cardiac valve replacement. In-hospital mortality is very high. The guidelines of the Society of Infectious Pathology of French Language (SPILF) for the prevention of infective endocarditis of at-risk patients were updated in 2002. They recommended a bi annual oral health follow-up and antibiotic prophylaxis for invasive sequences of care for high-risk patients. The objective of the study was to assess the application of the guidelines. METHODS: Using databases and medical files of self employed persons insured by compulsory national health insurance in the Alps, we studied from January 2001 to December 2002 the oral ambulatory follow-up of patients with permanent health insurance for valvular heart disease or congenital heart disease. RESULTS: Among the 260 patients with valvular heart disease or congenital heart disease, 125 (48.1%) presented medical claims for at least one dental treatment. Antibiotic prophylaxis was found in 15.8% of the invasive sequences of care for high-risk patients. The prescribed antibiotic was amoxicillin in 90% of the cases. CONCLUSION: Guidelines are seldom applied. Public and practitioner awareness should be improved to promote oral health: patients should regularly consult their dentist, and practitioners should follow the guidelines for the prevention of infective endocarditis. PMID- 16353516 TI - [Regression models for crude and relative survival: a comparative review]. AB - BACKGROUND: Statistical analysis of lifetime data is frequently used in the biomedical area. Our objective was to present a comparative review of the different regression models according to the survival concept (crude survival or relative survival) in order to express guidelines. METHODS: From a methodological point of view, we compared a regressive crude survival model (Cox model) and regressive relative survival models for grouped data (Hakulinen and Tenkanen) and for individuals data (Esteve et al.). We illustrated our work with an analysis of survival data of 3,355 incident cases of breast cancer identified by a hospital registry. Comportment of the models was studied in situation where censors rates ranged from 31.7 to 96.5%. RESULTS: Because relative survival analysis takes into account natural mortality, the risk of death was smaller for women older than 50 years than for women aged from 35 to 49; this was not demonstrated in the crude survival analysis (Cox). Estimations obtained from Cox model were more accurate than those obtained from both studied regressive relative survival models. Estimations obtained from Esteve et al. model were not very different from those obtained from Hakulinen and Tenkanen model and they were more accurate. CONCLUSION: By definition, analysis of relative survival is more appropriate to estimate survival to a specific cause of death. It is preferable to use models based on individual estimation when data set is small or when the number of individual per strata is small. PMID- 16353517 TI - [Tuberculosis and health care services in Burkina Faso]. PMID- 16353518 TI - Leprosy elimination in India--so near. PMID- 16353519 TI - Final push for elimination of leprosy in India. PMID- 16353520 TI - A study on the impact of FD-MDT on 200 leprosy patients. AB - A study was carried out from June 1999 to June 2001 to assess the impact of fixed duration multidrug therapy (FD-MDT) in newly detected cases of leprosy in terms of clinical and neurological improvement and changes in the bacillary index of skin smear for AFB. 200 new leprosy cases (both PB & MB) were started on FD-MDT. Of these 200 cases, 16 were of pure neuritic leprosy. After treatment, out of 184 cases with typical skin lesions of leprosy, all 26 PB cases showed inactivity of skin lesions, and, of the remaining 158 MB cases, 40.5% showed inactivity and 59.5% showed complete resolution of skin lesions. Out of 68 skin smear-positive cases, 42 cases with a BI of < or = 3 became smear-negative, while others showed gradual fall in the BI. Such heavily bacilliferous cases were continued with treatment for 1 more year to prevent relapse. As FD-MDT alone does not cure established sensory and motor impairment, it did not show any change in 19% of the patients presented with permanent sensory motor disturbance. FD-MDT prevents progression of sensory/motor disturbance. PMID- 16353521 TI - Integration of leprosy control into general health care system: observations from a state with low endemicity. AB - The study was undertaken as part of operational research to assess the level of integration of leprosy services into general health care system in 24 low or moderately endemic states/union territories by the Ministry of Health and Family Welfare, Government of India. Himachal Pradesh was one of the nine randomly selected states for the study. Out of the 12 districts in the State, 2 were selected randomly for the study. In each of the selected districts, 8 health facilities (that included a district hospital, an urban hospital/urban health centre, an Employees' State Insurance Hospital, a community health centre and a primary health centre) and 9 sub-centres were surveyed. Selection was done randomly at each stage. Data were collected on training in leprosy of general health care staff, availability of drugs for MDT in the system and maintenance of leprosy records by the staff of the system. The study showed mixed results. About half (53.2%) of the existing medical officers, 83.9% of health supervisors and 96.8% of multi-purpose workers were trained in leprosy. But only 31.3% of medical officers were able to diagnose leprosy and most of them were relying on vertical staff and skin specialists for confirmation. MDT services were provided by 20% of rural and 66.7% of urban health facilities that were acting as treatment centres. None of the health facilities had 3 months' stock of all types of blister packs, as per the guidelines of the Government of India. None of the sub-centres was involved in MDT delivery. However, reporting as per SIS formats was universal. The study emphasized the need for training and better management of MDT drug stock. PMID- 16353522 TI - Leprosy situation in the slums of Agra City--epidemiological findings. AB - The present article is the result of a study of the leprosy caseload in Agra City and is based on a house-to-house survey conducted during April-July 2003 in 5 areas. During the survey 198,150 persons were examined, and 287 cases were detected, giving a prevalence rate of 14.5/10,000. A majority of them (92%) were new cases, detected and diagnosed for the first time. The patient load was found to be unevenly distributed with comparatively more number of patients in areas such as Jamuna Kinara, Shah Ganj and Lohamandi. Among the 264 newly detected cases, 14.8% were of MB type. Overall deformity of grade > or = 2 was seen in 2.8% of patients--0.4% in PB and significantly high at 18% in MB leprosy. The observations reveal that leprosy is endemic in slum areas of Agra City. PMID- 16353523 TI - Leprosy among adolescents in Kolkata, India. AB - Leprosy, manifesting during adolescence when significant physical and emotional changes are taking place, poses further stress and strain both on the individual and on the family. Based on hospital records, focus group discussions and in depth interviews, data on 258 adolescent leprosy patients seen at a leprosy referral hospital in Kolkata, India, are presented. The male-female sex ratio was 1.93:1, 56.6% were multibacillary patients and 13.2% had grade 2 disability. At the time of final follow up, 10% of PB and 33% of MB patients had already discontinued treatment. The commonest complication was reaction (14.5%). Adolescents were still dependent on their parents for health matters. Data obtained from questionnaires confirmed the role of social stigma in hiding, delay in starting of MDT and defaulting. Frequent hospital admissions resulted in loss of jobs and disruption of studies and caused psychological disturbances. It is critical to identify and treat adolescent leprosy on a priority basis. Health education and counselling programmes must be more focused and acceptable. Further research is necessary. PMID- 16353524 TI - Closure of simple heel ulcers by skin stretching. AB - A majority of heel ulcers, at least to begin with, extend to dermis or to the fat pad in its superficial part and an appropriate skin closure can heal these ulcers as most of the padding is in tact. Since the skin is adherent to the deeper structures with fibrous bands it has to be stretched or undermined (by cutting the fibrous bands) to close the wound without tension. 17 feet in 11 patients (10 males; one female) in the 12-54 year age-group were operated upon and followed up. Because skin is adherent to deeper tissues by fibrous septae, stretching of skin was planned to mobilize it for a tension-free closure. Of the 17 feet, 13 could be re-examined after 30 months or more. Most of the minor recurrences were seen in the first 6 months after surgery. Major recurrences were seen in 2 feet (one case). The suture line did not show hyperkeratosis and the scar merged well into the surrounding skin after one year. Available data suggest that simple heel ulcers can be made to heal with a good scar by skin-stretching and suture, and, by radiography of the foot, it is worth separating those cases in which ulcer is not extending deep involving calcaneum. The size of the ulcer in heel is important for the success of the operation. The procedure is not intended for big wounds (>15 mm in width). PMID- 16353525 TI - Dapsone-induced eosinophilic pneumonitis in a leprosy patient. AB - A leprosy patient with no prior history of respiratory complaints, developed symptoms of dry cough, fever and dyspnea after six weeks of therapy. Peripheral eosinophilia and radiological evidence of pulmonary interstitial infiltrates pointed towards the possibility of drug-induced eosinophilic pneumonitis. The results of relevant tests for other possible pathologies were normal. The resolution of symptoms without any intervention other than withdrawal of the drug and subsequent re-challenge proved dapsone to be the cause. PMID- 16353526 TI - Leprosy masquerading as Melkersson-Rosenthal syndrome. PMID- 16353527 TI - Differential diagnosis of pure neuritic leprosy. PMID- 16353528 TI - National IAL workshop on the role of dermatologists in leprosy eradication. PMID- 16353529 TI - Osseous response following resective therapy with piezosurgery. AB - A piezoelectric instrument vibrating in the ultrasonic frequency range was investigated for its potential use in periodontal resective therapy. The rate of postoperative wound healing (baseline and 14, 28, and 56 days after surgery) in a dog model following surgical ostectomy and osteoplasty was the marker used to compare the efficacy of this instrument (PS) with a commonly used carbide bur (CB) or a diamond bur (DB). The surgical sites treated by CB or DB lost bone, in comparison to baseline measurements, by the 14th day, while the surgical sites treated by PS revealed a gain in the bone level. By day 28, the surgical sites treated by all three instruments demonstrated an increased bone level and regeneration of cementum and periodontal ligament. However, by day 56, the surgical sites treated by CB or DB evidenced a loss of bone, versus a bone gain in the PS-treated sites. Thus, it appears that PS provided more favorable osseous repair and remodeling than CB or DB when surgical ostectomy and osteoplasty procedures were performed. Therefore, PS could be regarded as being efficacious for use in osseous surgery. PMID- 16353530 TI - Sinus augmentation utilizing anorganic bovine bone (Bio-Oss) with absorbable and nonabsorbable membranes placed over the lateral window: histomorphometric and clinical analyses. AB - The purpose of the present study, which used anorganic bovine bone (Bio-Oss) with and without autogenous bone as the augmentation material, was to compare the results of sinus elevation performed without a membrane (control) with the results of sinus elevation performed with either a short-term bioabsorbable membrane (Bio-Gide) or a nonabsorbable membrane (Gore-Tex) with regard to both vital bone formation and implant survival. Sinus lifts were performed on 51 patients (38 unilateral, 13 bilateral) with the delayed placement of 135 implants. Histomorphometric data were obtained at the time of implant placement, 6 to 10 months following the grafting procedure. Vital bone formation was 17.6%, 16.9%, and 12.1%, respectively, for the Bio-Gide, Gore-Tex, and no membrane groups. Of the 135 implants placed there were 3 failures (2 Bio-Gide, 1 Gore Tex). There was no significant difference between the membrane groups as to vital bone formation and implant survival. PMID- 16353531 TI - Full-thickness flap/subepithelial connective tissue grafting with intramarrow penetrations: three case reports of lingual root coverage. AB - Three case reports are presented that demonstrate the use of full-thickness flap/subepithelial connective tissue grafting for root coverage on the lingual surfaces of the mandibular anterior teeth. This is accomplished using an envelope full-thickness flap technique with intramarrow penetrations at the recipient site. Miller Class I, II, and III gingival recession defects and gingival perforation defects were treated. Complete root coverage was achieved in two Miller Class I gingival recession defects, in one Miller Class II gingival recession defect, and in two gingival perforation defects in areas that exhibited no radiographic evidence of bone loss. Partial root coverage was achieved in two Miller Class III gingival recession defects in an area that exhibited radiographic evidence of bone loss. Although the majority of the exposed root surface was covered in these two Miller Class III defects, about 1 mm of root surface remained exposed, which seemed to closely correspond to the amount of bone loss that was noted radiographically. A grafting technique has been presented that can be used to restore the functional properties of the lingual gingiva of the mandibular anterior teeth by repairing gingival defects and re establishing the continuity and integrity of the zone of keratinized gingiva. Our clinical impression is that this has made it easier for the three patients presented in this report to maintain the lingual surfaces of the mandibular anterior teeth with routine oral hygiene measures. PMID- 16353532 TI - Effects of application of platelet releasate in periodontal regeneration therapy. AB - The alpha granules of platelets contain various growth factors, which display in vitro and in vivo activities known to be important in wound healing. Biologically active proteins from platelets include platelet-derived growth factor, transforming growth factor-beta, and insulin-like growth factor, as well as other less well-described angiogenic and differentiated protein factors. The purpose of this study was to evaluate histologically and histometrically the new tissue formation on furcation treatment with platelet-derived factor releasate (PR) in beagle dogs. Class II furcations were created in the mandibular premolars of eight adult male beagle dogs. Periodontal regenerative treatments were then performed using collagen sponge graft material with and without the topical application of PR. PR was prepared fresh from blood drawn from dogs before treatments. A histologic evaluation of the effect on new tissue formation was then performed by comparing periodontal tissue regeneration of sites treated with and without PR. Four and 12 weeks after the flap operations, histologic sections were processed and histologically analyzed. The evaluated parameters were bone, cementum and connective tissue attachment regeneration, length of epithelium, resorption, and ankylosis. Histologically evaluated, the regeneration of new cementum was greater at the collagen sponge with PR sites compared to the control sites throughout the entire research period (P < .01). Four and 12 weeks after the flap operations, the amount of new bone in the sites treated with PR was greater than that in the control group (P < .05). These results suggest that collagen sponge graft material with PR promotes new attachment on the periodontal tissue regeneration treatments. PMID- 16353533 TI - Further data on the predictability of the indirect sinus elevation procedure used with short, sintered, porous-surfaced dental implants. AB - The object of this report was to provide further data supporting the use of short (primarily 7-mm-long) dental implants with a sintered, porous-surface geometry to treat the posterior maxilla using the indirect, osteotome-mediated, localized sinus elevation procedure. Records were available for 104 Endopore implants (Innova) in 70 patients, for whom the majority of implants had been placed in the location of the maxillary first molar. The mean initial subantral bone height before implant placement was 4.2 mm, with a range of 2 to 6.7 mm, and all implants were placed using hand osteotomes and a graft of bovine hydroxyapatite. After an average time in function of 3.14 years, only two implants had been lost, both as a result of unusual circumstances. It is concluded that the use of short, sintered, porous-surfaced implants and localized indirect sinus elevation is a predictable and minimally invasive approach to manage the posterior maxilla with minimal preoperative subantral bone height. PMID- 16353534 TI - Acellular dermal matrix allograft used alone and in combination with enamel matrix protein in gingival recession: histologic study in dogs. AB - Gingival recession was created in six mongrel dogs. The dogs were divided into two groups based on treatment: group 1--AlloDerm only, group 2--AlloDerm + Emdogain. The histologic results were compared. At the end of the study, the mean values were, for groups 1 and 2, respectively: 0.06 and 0.32 mm for cementum regeneration; -0.75 and -0.86 mm for bone regeneration; -2.15 and -3.11 mm for attachment level; and 4.90 and 5.51 mm for defect extent. The epithelial formation parameter was 2.88 mm in group 1 and 2.15 mm in group 2, which was a statistically significant difference. It could be concluded that Emdogain did not result in beneficial effects when associated with AlloDerm. PMID- 16353535 TI - Bond integrity of dental core materials to esthetic intraradicular posts. AB - Bond strengths between various materials for endodontic post-core buildups are unknown. The study determined the diametral tensile strength of three esthetic post systems bonded directly or indirectly to four core materials, and subsequent scanning electron microscopy was performed on the fractures. Cylindric core buildup was made around the parallel-sided section of the posts using a custom made jig. Strength was tested after 14 days. A significant difference was found between the control group and the other groups (P = .0001). SEM observations were in agreement with the results. Apart from the control group, the results suggested that directly and indirectly performed post-core buildups have similar diametral tensile strength values. PMID- 16353536 TI - Histologic analysis of a failing three-part dental implant: a human case report. AB - Two failing implants were removed for prosthetic reasons from the maxilla of a 60 year-old woman. The implants were clinically immobile but presented clear signs of peri-implant mucositis and bone destruction. One of the two implants was harvested together with the surrounding bone and analyzed histologically. The implant was threaded and consisted of three distinct components screwed together in a telescopic fashion. Histologically, bone loss reached the fourth thread, while apical to the fourth thread, osseointegration between the host bone and the implant surface was recognizable. The implant parts were not completely seated into each other, and the resulting gap was colonized by host bone. Newly formed alveolar bone penetrated deep into the implant body cavity and appeared similar in nature to the alveolar bone surrounding the implant. Histologic findings are discussed in reference to the ability of such an implant to withstand biomechanical loading over time. PMID- 16353537 TI - A comparison of the clinical and histologic crestal bone level measurements adjacent to dental implants in the baboon. AB - The purpose of this study was to compare clinical probing measurements with the histologic alveolar bone level in an established baboon model. Implant sites were prepared by removal of all premolars and first molars in 10 adult female baboons (Papio anubis), a total of 120 sites. The resulting edentulous ridges were allowed to heal for 6 weeks. One implant was placed in each site and allowed to heal, after which the implants were removed en bloc. The animals were not sacrificed. Clinical probing from the implant platform to the crest of the bone was performed at the buccal and lingual aspects of each implant at the time of placement and prior to section removal. All measurements were taken with a calibrated probe by a single examiner (ARV). Sections were cut in a buccolingual plane through the long axis of each implant. Histologic slides were digitized for data collection, and histologic measurements were taken at the buccal and lingual aspects from the implant platform to the crest of the bone. The results of the study showed that there were no significant differences between the clinical and histologic measurements. The authors concluded that clinical measurements around dental implants are as reliable as histologic measurements and that the tip of the probe extended to the alveolar bone every time the implant was probed in this model. PMID- 16353538 TI - [Why not hematology?]. PMID- 16353539 TI - [Clinical and epidemiological features of leptospirosis in Bolivar state, Venezuela. Comparison of diagnostic methods: LEPTO-Dipstick and plaque macroscopic agglutination test]. AB - Leptospirosis is caused by numerous serovars of Leptospira interrogans. The infection is acquired through the contact of softened skin, mucous and conjunctive with the urine of animals and humans infected by animal carriers. The aim of this study was to determine the clinical and epidemiological features of leptospirosis in Bolivar state Venezuela and to compare the sensibility and specificity of the LEPTO-Dipstick, with the test of macroscopic agglutination in plate with the termorresistent antigen (TR). Thirty one sample of serums were processed of patients with leptospirosis clinic, admitted in the Hospital Universitario "Ruiz y Paez" of Ciudad Bolivar during 12 months. Detection IgM antibodies of anti-Leptospiras was carried out by means of LEPTO-Dipstick and antigen TR. The Microscopic Agglutination Test (MAT) was also, carried out. The general prevalence of leptospirosis in patients who attended the "Ruiz y Paez" Hospital was of 80.6% (n = 25) by means of the determination of the termorresistent antigen. The most frequent serovars identified in Bolivar State, were the serovars: icterohaemorrhagiae, copenhageni (21.3%), autummalis and australis (12.8%). TR antigen and LEPTO Dipstick had a sensibility of 80% and a specificity of 25%. The agreement between both methods was null (Kappa: -0.2). Bolivar state has a high leptospirosis prevalence and the infection should be discarded in those patients with long-term fever and risk factors for the illness. PMID- 16353540 TI - [Phenotypic characterization of species of Malassezia in healthy skin of an university student population]. AB - The yeasts of the Malassezia genus are part of the normal skin of man and other vertebrates. The description of new species for this genus has induced on their study in several countries. For this reason, is important to do research in order to get epidemiologic data about Malassezia species in tropical countries like Venezuela, where the new Malassezia species have not been reported. This study was made on healthy skin of a university student population. The samples were taken from different body areas and inoculated in Dixon modified medium and Sabouraud dextrose agar medium e incubated at 32 degrees C. The identification was achieved following the key of species described by Gueho et al. and the tween diffusion test proposed by Guillot et al. In this investigation the isolated specie was M. furfur, corresponding the major positive percentage to the age group from 16 to 20 years old (66.7%). The presence of Malassezia was predominant on shoulder and chest (33.3% and 26.6%). There were not found significant differences between the evaluated groups, as was demonstrated by the applied statistical tests, exact Fisher's test and Chi square test. It is important to continue the investigations on other age groups, in order to establish the prevalent species in our region and evaluate their pathogenic potential. PMID- 16353541 TI - [Evaluation of K-ras 12 oncogene mutations in Venezuelan patients with Helicobacter pylori infection]. AB - This work evaluated the infection of H. pylori in the different gastric pathologies and its association with the oncogen K-ras 12. Endoscopy was performed in 62 patients and 3 biopsies from the antral region were taked and used for the histological diagnostic, PCR, and point mutations determination. The results showed a high incidence of H. pylori infection in patients with active chronic gastritis (AcCG) 90%, chronic atrophic gastritis (AtCG) 70%, intestinal metaplasia (IM) 67%, dysplasia (D) 83%, and decrease in in gastric cancer (GC) 33%. Evaluation of the oncogen K-ras 12 showed that 68% of the patients presented mutations in the different analyzed amino acids. In the 12 codon of the K-ras gene, we observed simple point mutations and combination in the same sample in different gastric pathologies. In AcCG samples were detected the greater number of mutations. A decrease of the point mutations were observed in the progression stages to gastric cancer. The presence of these specific mutations would be tumor markers and it determine the possible development of gastric tumors. PMID- 16353542 TI - [Homocysteinemia and its relationship with the methylentetrahydrofolate reductase polymorphism in various ethnic groups from western Venezuela]. AB - The prevalence of hyperhomocysteinemia and C677T MTHFR polymorphism was studied in various ethnic groups from Western Venezuela (60 Wayuu Indians, 42 italian immigrants and 77 Venezuelan mestizos) in relation with the prevalence of hyperhomocysteinemia and the C677T MTHFR polymorphism. Homocysteinemia was determined by polarized fluorescence immunoassay in an IMX system, serum folate was measured by radioimmunoanalysis and the MTHFR genotype was determined by PCR and restriction analysis. Hyperhomocysteinemia was defined as a value over 2 SD above the mean value for normal MTHFR (CC677) in each group. The prevalence of MTHFR variants (C677T and 677TT) was elevated in all ethnic groups (78% among the wayuu, 76% among Italians and 63% among mestizos) with a significant association between the concentrations of homocysteine and the levels of serum folate among the wayuu (p < 0.0001) and the mestizos (p < 0.001) only. Hyperhomocysteinemia was associated with MTHFR variants in 23% of the wayuu (OR: 6.17, CI 95: 0.74 51.36), 9.5% of the Italians (OR: 0.93, CI 95: 0.085-10.10) and 20.7 of the Venezuelans mestizos (OR: 5.2, CI 95: 1.08-24.90, p > 0.03). There was no relationship between hyperhomocysteinemia and folate deficiency in any of the groups studied. In conclusion, despite a high prevalence of C677T MTHFR variants in these ethnic groups of western Venezuela, the lack of no evidence of hyperhomocysteinemia combined with folate deficiency may imply that the nutritional status of these groups plays an important role in the control of hyperhomocysteinemia as a risk factor for cardiovascular disease. PMID- 16353543 TI - [Importance of IgG anti-CagA antibodies of Helicobacter pylori in Venezuelan patients with gastric diseases]. AB - Helicobacter pylori is one of the most common bacterial infections worldwide. It is associated with chronic gastritis, peptic ulcer disease and constitutes a major risk factor for gastric adenocarcinoma and lymphoma. The aim of this study was to evaluate the specific serologic immunoglobulin G (IgG) response to whole cells proteins, CagA and urease antigens of Helicobacter pylori in a Venezuelan population. We evaluated 66 patients from the Hospital Universitario de Caracas, attending in the gastroscopy service. H. pylori infection was detected by culture and rapid urease test. IgG antibodies against, CagA and ureases were tested by enzyme-linked immunosorbent assay method using highly purified recombinant antigens. We demonstrated the presence of H. pylori in 48/66 (72.7%), by culture and rapid urease test. We found a seroprevalence of 45 (68%) to whole cells, 34/66 (51%) to CagA and 18/66 (27%) to urease. The positive rates of CagA antibodies in patients with gastric ulcer, gastric cancer and chronic gastritis were 87.8%, 77.7% y 40.8% respectively. The serum antibodies anti-CagA were similar between peptic ulcer disease and gastric cancer patients. PMID- 16353544 TI - A system for the detection and typing of human papillomavirus of the lower genital tract: in situ hybridization screening and polymerase chain reaction confirmation. AB - We developed a simplified and non-isotopic in situ hybridization procedure for the detection of Human Papillomavirus (HPV) in routine Papanicolaou cervical smears. The assay involves one oligonucleotide (malignant probe) which recognizes high risk HPV 16 and 18, and another which detects HPV 6 and 11 (benign probe). We adapted the system to fulfill the requirements of gynecologists and cytologists, assimilating their protocols and simplifying the in situ hybridization assay. When we compared the detection levels reached by the in situ hybridization versus a ladder PCR assay in 156 clinical samples, original designed for this work, the kappa coefficient between both methods is 0.945, indicating a strong agreement between the ISH and the PCR assays. PMID- 16353545 TI - [Some immunological aspects in the reaccional states of Hansen disease]. AB - The crucial clinical problem in leprosy is represented by episodes of intense inflammation that produce nerve damage. When Mycobacterium leprae has been eliminated by means of antibiotics, the death of the bacteria is not a complete solution to the damage caused in nerves. Two of the more frequent immunopathological phenomena in Hansen's disease are Type I, reactions, known as Reversal Reactions (RR), and Type II reactions, of which the most frequent and well known are those called Erythema Nodosum Leprosum (ENL). Type II reactions have been defined as an immunologic complication in multibacillary patients. Both types of reactions are accompanied mainly by the increase of pro-inflammatory cytokines TNF-alpha, IL-1, IL-2, IL-4, IL-6, IL-8, INF-gamma, IL-10, IL-12, among others. In a retrospective analysis of a population of 150 patients of the Central Service of Dermatology, Institute of Biomedicine, where these patients were subjected to Multidrug Therapy (MDT) and MDT + inmunotherapy (IMT), both groups presented type II reactions, but the group that additionally received inmunotherapy also presented type I reactions. In the group of patients that presented ENL, it appeared to be associated with the large amount of bacilli in their lesions. PMID- 16353546 TI - [Cell immunity response in human pulmonary tuberculosis. Review]. AB - Human pulmonary tuberculosis (TB) is a worldwide public health problem, which is caused by Mycobacterium tuberculosis. It is a fact that one third of the world's population is infected with this mycobacteria, however, only a minority of people infected by M. tuberculosis may develop a clinical disease. In general, about 90% have their bacilli under control in a latent state throughout their lives by means of their immune responses. About 5% will develop primary progressive TB and the remaining 5% will develop the disease in the later stages of their lives, which is known as reactivation or post-primary TB. In resistant individuals, control of the infection mainly requires development of a Th1 cell immunity response. This type of response involves participation of alveolar macrophages and T CD4+, CD8+ and T gammadelta lymphocytes, and production of cytokines such as IL-2, IFN-gamma, IL-12, IL-18 and TNF-alpha, as well as chemokines such as RANTES, MCP-1, MIP-1alpha and IL-8 which play an important role in the migration of different cell subpopulations to the infection site for the formation of granulome. In addition, the role of "natural killer" (NK) cells, along with epitelial cells, is essential as part of the innate immune response. PMID- 16353547 TI - Exopolysaccharide structure is not a determinant of host-plant specificity in nodulation of Vicia sativa roots. AB - Exopolysaccharide (EPS)-deficient strains of the root nodule symbiote Rhizobium leguminosarum induce formation of abortive infection threads in Vicia sativa subsp. nigra roots. As a result, the nodule tissue remains uninfected. Formation of an infection thread can be restored by coinoculation of the EPS-deficient mutant with a Nod factor-deficient strain, which produces a similar EPS structure. This suggests that EPS contributes to host-plant specificity of nodulation. Here, a comparison was made of i) coinoculation with heterologous strains with different EPS structures, and ii) introduction of the pRL1JI Sym plasmid or a nod gene-encoding fragment in the same heterologous strains. Most strains not complementing in coinoculation experiments were able to nodulate V. sativa roots as transconjugants. Apparently, coinoculation is a delicate approach in which differences in root colonization ability or bacterial growth rate easily affect successful infection-thread formation. Obviously, lack of infection-thread formation in coinoculation studies is not solely determined by EPS structure. Transconjugation data show that different EPS structures can allow infection thread formation and subsequent nodulation of V. sativa roots. PMID- 16353548 TI - Identification of a protein from rust fungi transferred from haustoria into infected plant cells. AB - The formation of haustoria is one of the hallmarks of the interaction of obligate biotrophic fungi with their host plants. In addition to their role in nutrient uptake, it is hypothesized that haustoria are actively involved in establishing and maintaining the biotrophic relationship. We have identified a 24.3-kDa protein that exhibited a very unusual allocation. Rust transferred protein 1 from Uromyces fabae (Uf-RTP1p) was not only detected in the host parasite interface, the extrahaustorial matrix, but also inside infected plant cells by immunofluorescence and electron microscopy. Uf-RTP1p does not exhibit any similarity to sequences currently listed in the public databases. However, we identified a homolog of Uf-RTP1p in the related rust fungus Uromyces striatus (Us RTP1p). The localization of Uf-RTP1p and Us-RTP1p inside infected plant cells was confirmed, using four independently raised polyclonal antibodies. Depending on the developmental stage of haustoria, Uf-RTP1p was found in increasing amounts in host cells, including the host nucleus. Putative nuclear localization signals (NLS) were found in the predicted RTP1p sequences. However, functional efficiency could only be verified for the Uf-RTP1p NLS by means of green fluorescent protein fusions in transformed tobacco protoplasts. Western blot analysis indicated that Uf-RTP1p and Us-RTP1p most likely enter the host cell as N-glycosylated proteins. However, the mechanism by which they cross the extrahaustorial membrane and accumulate in the host cytoplasm is unknown. The localization of RTP1p suggests that it might play an important role in the maintenance of the biotrophic interaction. PMID- 16353549 TI - Fusarium oxysporum gas1 encodes a putative beta-1,3-glucanosyltransferase required for virulence on tomato plants. AB - Glycosylphosphatidylinositol-anchored (beta)-1,3-glucanosyltransferases play active roles in fungal cell wall biosynthesis and morphogenesis and have been implicated in virulence on mammals. The role of beta-1,3-glucanosyltransferases in pathogenesis to plants has not been explored so far. Here, we report the cloning and mutational analysis of the gas1 gene encoding a putative beta-1,3 glucanosyltransferase from the vascular wilt fungus Fusarium oxysporum. In contrast to Candida albicans, expression of gas1 in F. oxysporum was independent of ambient pH and of the pH response transcription factor PacC. Gene knockout mutants lacking a functional gas1 allele grew in a way similar to the wildtype strain in submerged culture but exhibited restricted colony growth on solid substrates. The restricted growth phenotype was relieved by the osmotic stabilizer sorbitol, indicating that it may be related to structural alterations in the cell wall. Consistent with this hypothesis, deltagas1 mutants exhibited enhanced resistance to cell wall-degrading enzymes and increased transcript levels of chsV and rho1, encoding a class V chitin synthase and a small monomeric G protein, respectively. The deltagas1 mutants showed dramatically reduced virulence on tomato, both in a root infection assay and in a fruit tissue invasion model, thus providing the first evidence for an essential role of fungal beta-1,3-glucanosyltransferases during plant infection. PMID- 16353550 TI - Evolution of an avirulence gene, AVR1-CO39, concomitant with the evolution and differentiation of Magnaporthe oryzae. AB - The significance of AVR1-CO39, an avirulence gene of the blast fungus corresponding to Pi-CO39(t) in rice cultivars, during the evolution and differentiation of the blast fungus was evaluated by studying its function and distribution in Pyricularia spp. When the presence or absence of AVR1-CO39 was plotted on a dendrogram constructed from ribosomal DNA sequences, a perfect parallelism was observed between its distribution and the phylogeny of Pyricularia isolates. AVR1-CO39 homologs were exclusively present in one species, Pyricularia oryzae, suggesting that AVR1-CO39 appeared during the early stage of evolution of P. oryzae. Transformation assays showed that all the cloned homologs tested are functional as an avirulence gene, indicating that selection has maintained their function. Nevertheless, Oryza isolates (isolates virulent on Oryza spp.) in P. oryzae were exceptionally noncarriers of AVR1-CO39. All Oryza isolates suffered from one of the two types of known rearrangements at the Avr1 CO39 locus (i.e., G type and J type). These types were congruous to the two major lineages of Oryza isolates from Japan determined by MGR586 and MAGGY. These results indicate that AVR1-CO39 was lost during the early stage of evolution of the Oryza-specific subgroup of P. oryzae. Interestingly, its corresponding resistance gene, Pi-CO39(t), is not widely distributed in Oryza spp. PMID- 16353551 TI - Expression profiling soybean response to Pseudomonas syringae reveals new defense related genes and rapid HR-specific downregulation of photosynthesis. AB - Transcript profiling during susceptible (S) and hypersensitive response associated resistance (R) interactions was determined in soybean (Glycine max). Pseudomonas syringae pv. glycinea carrying or lacking the avirulence gene avrB, was infiltrated into cultivar Williams 82. Leaf RNA was sampled at 2, 8, and 24 h postinoculation (hpi). Significant changes in transcript abundance were observed for 3,897 genes during the experiment at P < or = 0.000005. Many of the genes showed a similar direction of increase or decrease in abundance in both the S and R responses, but the R response generally showed a significantly greater degree of differential expression. More than 25% of these responsive genes had not been previously reported as being associated with pathogen interactions, as 704 had no functional annotation and 378 had no homolog in National Center for Biotechnology Information databases. The highest number of transcriptional changes was noted at 8 hpi, including the downregulation of 94 chloroplast-associated genes specific to the R response. Photosynthetic measurements were consistent with an R-specific reduction in photosystem II operating efficiency (phiPSII) that was apparent at 8 hpi for the R response with little effect in the S or control treatments. Imaging analyses suggest that the decreased phiPSII was a result of physical damage to PSII reaction centers. PMID- 16353552 TI - A ClC chloride channel homolog and ornithine-containing membrane lipids of Rhizobium tropici CIAT899 are involved in symbiotic efficiency and acid tolerance. AB - Rhizobium tropici CIAT899 is highly tolerant to several environmental stresses and is a good competitor for nodule occupancy of common bean plants in acid soils. Random transposon mutagenesis was performed to identify novel genes of this strain involved in symbiosis and stress tolerance. Here, we present a genetic analysis of the locus disrupted by the Tn5 insertion in mutant 899-PV9, which lead to the discovery of sycA, a homolog of the ClC family of chloride channels and Cl-/H+ exchange transporters. A nonpolar deletion in this gene caused serious deficiencies in nodule development, nodulation competitiveness, and N2 fixation on Phaseolus vulgaris plants, probably due to its reduced ability to invade plant cells and to form stable symbiosomes, as judged by electron transmission microscopy. A second gene (olsC), found downstream of sycA, is homologous to aspartyl/asparaginyl beta-hydroxylases and modifies two species of ornithine-containing lipids in vivo, presumably by hydroxylation at a still unknown position. A mutant carrying a nonpolar deletion in olsC is symbiotically defective, whereas overexpressed OlsC in the complemented strain provokes an acid sensitive phenotype. This is the first report of a ClC homolog being essential for the establishment of a fully developed N2-fixing root nodule symbiosis and of a putative beta-hydroxylase that modifies ornithine-containing membrane lipids of R. tropici CIAT899, which, in turn, are contributing to symbiotic performance and acid tolerance. PMID- 16353553 TI - Resistance quantitative trait loci originating from Solanum sparsipilum act independently on the sex ratio of Globodera pallida and together for developing a necrotic reaction. AB - Plant resistance to nematodes is related to the ability of the host to reduce the development of nematode juveniles into females. Resistance to the potato cyst nematode (PCN) Globodera pallida, originating from the wild species Solanum sparsipilum, was dissected by a quantitative trait loci (QTL) approach. Two QTL explained 89% of the phenotypic variation. The QTL GpaV(s)spl on chromosome V displayed the major effect on the cyst number (coefficient of determination [R2] = 76.6%). It restricted G. pallida development to 16.2% of juveniles, 81.5% of males, and 2.3% of females. The QTL GpaXI(s)spl on chromosome XI displayed a lower effect on the cyst number (R2 = 12.7%). It restricted G. pallida development to 13.8% of juveniles, 35.4% of males, and 50.8% of females. Clones carrying both QTL restricted the nematode development to 58.1% juveniles, 41.1% of males, and 0.8% of females. We demonstrated that potato clones carrying both QTL showed a strong necrotic reaction in roots infected by nematodes, while no such reaction was observed in clones carrying a single QTL. This result underlines the importance to introgress together GpaV(s)spl and GpaXI(s)spl into potato cultivars, in order to reduce the density of this quarantine pest in soil and to decrease the risk of selecting overcoming G. pallida subpopulations. PMID- 16353554 TI - Nod factors alter the microtubule cytoskeleton in Medicago truncatula root hairs to allow root hair reorientation. AB - The microtubule (MT) cytoskeleton is an important part of the tip-growth machinery in legume root hairs. Here we report the effect of Nod factor (NF) on MTs in root hairs of Medicago truncatula. In tip-growing hairs, the ones that typically curl around rhizobia, NF caused a subtle shortening of the endoplasmic MT array, which recovered within 10 min, whereas cortical MTs were not visibly affected. In growth-arresting root hairs, endoplasmic MTs disappeared shortly after NF application, but reformed within 20 min, whereas cortical MTs remained present in a high density. After NF treatment, growth-arresting hairs were swelling at their tips, after which a new outgrowth formed that deviated with a certain angle from the former growth axis. MT depolymerization with oryzalin caused a growth deviation similar to the NF; whereas, combined with NF, oryzalin increased and the MT-stabilizing drug taxol suppressed NF-induced growth deviation. The NF-induced disappearance of the endoplasmic MTs correlated with a loss of polar cytoarchitecture and straight growth directionality, whereas the reappearance of endoplasmic MTs correlated with the new set up of polar cytoarchitecture. Drug studies showed that MTs are involved in determining root hair elongation in a new direction after NF treatment. PMID- 16353555 TI - PecS and PecT coregulate the synthesis of HrpN and pectate lyases, two virulence determinants in Erwinia chrysanthemi 3937. AB - Erwinia chrysanthemi strain 3937 is a necrotrophic bacterial plant pathogen. Pectinolytic enzymes and, in particular, pectate lyases play a key role in soft rot symptoms; however, the efficient colonization of plants by E. chrysanthemi requires additional factors. These factors include HrpN (harpin), a heat-stable, glycine-rich hydrophilic protein, which is secreted by the type III secretion system. We investigated the expression of hrpN in E. chrysanthemi 3937 in various environmental conditions and different regulatory backgrounds. Using lacZ fusions, hrpN expression was markedly influenced by the carbon source, osmolarity, growth phase, and growth substrate. hrpN was repressed when pectinolysis started and negatively regulated by the repressors of pectate lyase synthesis, PecS and PecT. Primer extension data and in vitro DNA-protein interaction experiments support a model whereby PecS represses hrpN expression by binding to the hrpN regulatory region and inhibiting transcript elongation. The results suggest coordinated regulation of HrpN and pectate lyases by PecS and PecT. A putative model of the synthesis of these two virulence factors in E. chrysanthemi during pathogenesis is presented. PMID- 16353556 TI - Expression levels of avrBs3-like genes affect recognition specificity in tomato Bs4- but not in pepper Bs3-mediated perception. AB - The tomato Bs4 disease resistance gene mediates recognition of avrBs4-expressing strains of the bacterial spot pathogen Xanthomonas campestris pv. vesicatoria to give a hypersensitive response (HR). Here, we present the characterization of the Bs4 promoter and its application for low-level expression of bacterial type III effector proteins in planta. Real-time polymerase chain reaction showed that Bs4 is constitutively expressed at low levels and that transcript abundance does not change significantly upon infection with avrBs4-containing xanthomonads. A 302-bp promoter fragment was found to be sufficient to promote Bs4 gene function. Previous studies have shown that high, constitutive in planta expression of avrBs3 (AvrBs3 and AvrBs4 proteins are 96.6% identical) via the Cauliflower mosaic virus 35S (35S) promoter triggers a Bs4-dependent HR whereas X. campestris pv. vesicatoria-mediated delivery of AvrBs3 into the plant cytoplasm does not. Here, we demonstrate that, when expressed under control of the weak Bs4 promoter, avrBs3 does not trigger a Bs4-dependent HR whereas avrBs4 does. In contrast, the pepper Bs3 gene, which mediates recognition of AvrBs3- but not AvrBs4-delivering xanthomonads, retains its recognition specificity even if avrBs4 was expressed in planta from the strong 35S promoter. Importantly, Bs4 promoter-driven expression of hax3, hax4 (two recently isolated avrBs3-like genes), avrBs3, and avrBs4 resulted in identical reactions as observed upon infection with X. campestris pv. vesicatoria strains that express the respective avr gene, suggesting that the protein levels expressed under control of the Bs4 promoter are similar to those that are translocated by the bacterial type III secretion system. PMID- 16353557 TI - Genetic analysis of developmentally regulated resistance to downy mildew (Hyaloperonospora parasitica) in Arabidopsis thaliana. AB - Although developmentally regulated disease resistance has been observed in a variety of plant-pathogen interactions, the molecular basis of this phenomenon is not well understood. Arabidopsis thaliana ecotype Columbia-0 (Col-0) expresses a developmentally regulated resistance to Hyaloperonospora parasitica isolate Emco5. Col-0 seedlings support profuse mycelial growth and asexual spore formation in the cotyledons. In contrast, Emco5 growth and reproduction is dramatically (but not completely) restricted in the first set of true leaves. Subsequent leaves exhibit progresssively increased resistance. This adult resistance is strongly suppressed by expression of the salicylic acid-degrading transgene NahG and by loss-of-function mutations in the defense-response regulators PAD4, NDR1, RAR1, PBS3, and NPR1. In contrast to Col-0, the Wassilewskija-0 (Ws-0) ecotype supports profuse growth of Emco5 at all stages of development. Gene-dosage experiments and segregation patterns indicate that adult susceptibility in Ws-0 is incomepletely dominant to adult resistance in Col-0. Genetic mapping in a Col x Ws F2 population revealed a major locus on the bottom arm of chromosome 5, which we named RPP31. Analysis of T-DNA insertion lines indicated that the Columbia allele of RPP8, though tightly linked to RPP31, is not necessary for adult resistance. PMID- 16353559 TI - Practice-based commissioning: GPs set to receive extra cash. PMID- 16353558 TI - Characterization of the binding of diarrheagenic strains of E. coli to plant surfaces and the role of curli in the interaction of the bacteria with alfalfa sprouts. AB - Diarrheagenic Escherichia coli were able to bind to plant surfaces, including alfalfa sprouts and open seed coats, and tomato and Arabidopsis thaliana seedlings incubated in water. The characteristics of the binding differed with the bacterial strain examined. Laboratory K12 strains of E. coli failed to show significant binding to any of the plant surfaces examined, suggesting that some of the genes present and expressed in pathogenic strains and absent or unexpressed in K12 strains may be required for binding to plants. When a plasmid carrying the mlrA gene (a positive regulator of curli biosynthesis) or a plasmid carrying the operons that encode the synthesis of curli (csgA-G) was introduced into K12 strains, the bacteria acquired the ability to bind to sprouts. CsgA mutants of an avian pathogenic E. coli and an O157:H7 strain showed no reduction in their ability to bind to sprouts. Thus, the production of curli appears to be sufficient to allow K12 strains to bind, but curli are not necessary for the binding of pathogenic strains, suggesting that pathogenic strains may have more than one mechanism for binding to plant surfaces. PMID- 16353560 TI - One in three PCTs in the red. PMID- 16353561 TI - Homeless people struggling to get care. PMID- 16353562 TI - Who's in charge: regulation heavyweights square up. PMID- 16353563 TI - On championing regeneration. PMID- 16353564 TI - Data briefing. Patient charges. PMID- 16353566 TI - Implementing NICE guidance. Get the message. AB - Take-up of some NICE guidance is often erratic. Payment by results will change the way implementation of NICE guidance is funded, but there is little clarity on how it will work locally. Variations are particularly pronounced in cancer drugs. PMID- 16353565 TI - Primary care. The real deal. AB - Primary care trusts will be able to retain some provision functions in the restructured NHS. Some PCTs, such as in Greater Manchester, have plans to identify and develop services with community providers. PMID- 16353567 TI - Implementing NICE guidance. Roundtable discussion. Talking cure. PMID- 16353568 TI - Implementing NICE guidance. Best practice. Think local. AB - Leadership, prioritisation and planning as much as 18 months in advance have helped implementation. Effective networks bringing together primary and secondary care make implementation more efficient. Interactive IT can be a powerful tool. PMID- 16353569 TI - Implementing NICE guidance. Implementation planning. Nice work if you can do it. PMID- 16353570 TI - On public health. PMID- 16353571 TI - [In memory of John Paul II]. PMID- 16353572 TI - [The challenge of nursing teachers training: contribution to changes in training practices]. PMID- 16353573 TI - [Communication in palliative care: transmission of more news]. PMID- 16353574 TI - [The role of schools in the evolution of nursing teaching: 1886-1930]. PMID- 16353575 TI - [Training: contribution to quality]. PMID- 16353576 TI - [Hydration and its effects on health]. PMID- 16353578 TI - [Seminary of the CICIAMS in Vienna and visit to Prague, Bratislava, Budapest, and Vienna]. PMID- 16353577 TI - [The sisters of Sao Vicente de Paulo in the United States of America]. PMID- 16353579 TI - Endocrine regulation of fetal adipose tissue metabolism in the pig: role of hydrocortisone. AB - Glucocorticoids have been shown to be essential for the excessive fat deposition and development of obesity in several animal models. This study was performed to characterize the role of glucocorticoids in the developmental regulation of adipose tissue metabolism. On day 70 of gestation, pig fetuses were hypophysectomized by micro-cauterization. Hypophysectomized fetuses were implanted subcutaneously with hydrocortisone pellets or received no hormone replacement. Fetuses were removed by laparotomy on day 90 of gestation. Additional fetuses were hypophysectomized on day 70, implanted with hydrocortisone pellets on day 90 and removed on day 105 of gestation. Several intact fetuses were also implanted subcutaneously with hydrocortisone pellets during this later gestational period. Serum cortisol concentrations were reduced in hypophysectomized pigs at both fetal ages and were restored to intact levels by hydrocortisone treatment. Hydrocortisone supplementation enhanced lipolytic response to isoproterenol in intact fetuses but failed to restore lipolytic response to isoproterenol in hypophysectomized animals at either fetal age. Hydrocortisone induced a slight increase in lipogenesis in hypophysectomized fetuses when administered from 70 to 90 days of gestation and a more dramatic increase when administered from days 90 to 105 of gestation. However, hydrocortisone had no effect on basal or insulin stimulated lipogenesis in intact fetuses when administered from days 90 to 105 of gestation. These results indicate that hydrocortisone may have a primary influence on adipose tissue metabolism during late fetal development only in the absence of inhibition from counterregulatory hormones of pituitary origin. PMID- 16353580 TI - Estimates of the prevalence of obesity in Canadian children. AB - Childhood obesity is becoming a topic of great con cern due to the rising prevalence of this condition in North America. Studies conducted in the United States have indicated that the prevalence of obesity has increased dramatically over the past few decades. The purpose of this study was to estimate the prevalence of obesity in Canadian children between the ages of 5 and 12 years by examining data from two national and two regional surveys The 85th percentiles of each of four anthropometric indices derived from large normative populations were used as diagnostic criteria for obesity. As expected, the resulting prevalences varied according to the criteria used. A significant increase in childhood obesity between the 1981 to 1988 national surveys was observed when the three indices which used skinfolds were applied Weight-for-height percentiles did not indicate an increase in obesity in these samples. Regional samples showed a less than expected prevalence of obesity among the middle-class children and a higher than expected rate among the inner city boys. It can be concluded that there is a need for a defined criteria for identifying obesity in children in order to avoid confusion resulting from the wide variation in estimates of prevalence resulting from different standards and measurements. Using adiposity-based criteria for obesity it was clearly evident that the prevalence of obesity has increased in Canadian children. PMID- 16353581 TI - Hypersensitivity of the corticotropic axis to the serotoninergic agent clomipramine in obese women. AB - Serotoninergic control of food intake has been shown to be abnormal in obese persons with a decrease in serotoninergic tone. The neuroendocrine effects of intravenous I.V. administration of clomipramine (CMI), a serotonin uptake inhibitor, were studied in normal-weight (n=7) and obese subjects before (n=12) and after (n=6) dietary restriction. Under double-blind, placebo-controlled conditions, a single 12.5 mg dose of CMI was administered. There was no difference in baseline values of prolactin (PRL), corticotropin (ACTH) and cortisol in non-obese controls, obese before and obese after weight loss. CMI led to significant increases of PRL, ACTH, and cortisol concentrations in the controls as well as the obese group. The ACTH and cortisol responses to CMI in obese subjects were somewhat greater than the responses in normal-weight subjects. The area under the curve AUC for ACTH after clomipramine was 6202 +/- 976 pg/ml x 150 minutes for tile obese before weight loss and 3274 +/- 512 pg/ml x 150 minutes for the controls and the difference was significant at the level of p=0.052. The cortisol peak value after clomipramine was 163.71 +/- 14.31 ng/ml in the non-obese and 214.66 +/- 12.59 ng/ml in the obese (p=0.025). However, there was no difference in the obese subjects before and after weight loss. These data support the assumption that obese women have an abnormal sensitivity to the serotoninergic control of the hypothalamic pituitary adrenal axis (HPA), and that a mild weight loss does not significantly modify their serotoninergic tone. PMID- 16353582 TI - Baseline predictors of success when comparing two treatments. AB - Since few medications are equally effective in all patients, physicians can maximize the risk/benefit ratio of therapy for their patients by limiting exposure based on baseline predictors of success. Traditional procedures typically evaluate the response of patients receiving the same treatment regimen without evaluating a comparator. However, when treatments are compared, such as in clinical trials, traditional procedures of identifying predictors must be modified to analyze the treatment effect on the primary outcome variable. We focus on clinical and statistical considerations that arise when developing baseline predictors through models which consider treatment differences. To illustrate an application of this method, we used data from 1,026 patients completing at least 6 months of double-blind therapy in clinical trials comparing fluoxetine (N=522) with placebo (N=504) for weight loss. Stepwise regression procedures were used to identify baseline variables which were predictive of a beneficial fluoxetine treatment effect on last-visit-carried-forward (LVCF) weight change. In this example, age, smoking activity, and uric acid concentration were the best baseline predictors of long-term treatment effect relative to LVCF weight change. Patients were more likely to achieve long-term benefit with fluoxetine if they were older, and/or were nonsmokers, and/or had high concentrations of uric acid at baseline. These predictors, developed through models keying on treatment effect, can be used to identify patients who are more likely to accrue benefits with active therapy beyond those expected with placebo therapy, thus enriching the treatment population so that a higher proportion of treated patients are successful. PMID- 16353583 TI - The visceral adiposity syndrome in Japanese-American men. AB - Japanese-Americans have an increased prevalence of non-insulin-dependent diabetes mellitus and coronary heart disease when compared to native Japanese. This increase has been associated with fasting hyperinsulinemia, hypertriglyceridemia, and low plasma levels of high-density lipoprotein (HDL) cholesterol. The purpose of this study was to examine the relationship of both visceral adiposity and insulin resistance to this metabolic syndrome and to the presence of a predominance of small, dense low-density lipoprotein (LDL) particles (LDL subclass phenotype B) that has been associated with increased atherogenic risk. Six Japanese-American men with non-insulin-dependent diabetes, each receiving an oral sulfonylurea, were selected. One or 2 nondiabetic Japanese-American men, matched by age and body mass index, were selected for each diabetic subject, giving a total of 9 nondiabetic men. Diabetic subjects had significantly higher fasting plasma glucose (p=0.0007) and lower insulin sensitivity (SI, p=0.018) using the minimal model technique than nondiabetic subjects matched for body mass index. Six men (2 with diabetes) had LDL phenotype A and 8 (4 with diabetes) had phenotype B. One nondiabetic subject had an intermediate low-density lipoprotein pattern. Significantly greater amounts of intra-abdominal fat (p=0.045) measured by computed tomography were found in the men with phenotype B while fasting insulin (p=0.070) and triglycerides (p=0.051) tended to be higher. free fatty acids (r=0.677), LDL density (relative flotation rate, r=-0.803), and plasma HDL cholesterol (r=-0.717). SI was significantly correlated only with plasma free fatty acids (r=-0.546) and tended to be correlated with hepatic lipase activity (r=-0.512, p=0.061). In conclusion, these observations indicate that in non-obese Japanese-American men, the metabolic features of the so-called insulin resistance syndrome, including LDL phenotype B, are more strongly correlated with visceral adiposity than with SI. It may therefore be more appropriate to call this the visceral adiposity syndrome. Although questions concerning mechanisms still remain, we postulate that visceral adiposity plays a central role in the development of many of the metabolic abnormalities, including LDL subclass phenotype B, that occur in this metabolic syndrome. PMID- 16353584 TI - The quantitative relationship between "the metabolic syndrome" and abdominal obesity in women. AB - Abdominal obesity is closely associated with the presence of metabolic risk factors and elevated blood pressure in selected materials. This has, however, never been analyzed quantitatively in a non-selected cohort. Therefore, in a population-based study of 1462 Swedish women, four selected risk factors for non insulin dependent diabetes mellitus (NIDDM) and cardiovascular disease (CVD), serum triglycerides, blood glucose and systolic blood pressure and also serum insulin in a subsample, were examined in relation to regional and overall obesity. This was performed by subdividing the age adjusted sample into quintiles of waist to hip circumference ratio (WHR) or body mass index (BMI) as indicators of abdominal distribution of body fat and overall obesity, respectively. The risk factors serum triglycerides, blood glucose, blood pressure and serum insulin were defined as being elevated when the value of the risk factor was higher than the mean plus one or two standard deviations of the total age-adjusted cohort. The percentage of women with elevated risk factors according to this definition was then calculated in each of these quintiles. Having a risk factor which was elevated according to the definition was significantly correlated to WHR and BMI (p<0.0001) independent of age. The presence of one or several of these elevated risk factors was clearly higher than expected in the fifth quintile of WHR, and to a lesser extent in the fifth quintile of BMI while this was not the case in the lower quintiles of WHR and BMI. When studying the combination of the WHR and BMI, the presence of risk factors higher than the mean plus two standard deviations increased gradually with WHR in all five quintiles of BMI. A significant association was observed between WHR and presence of risk factors independent of BMI (p<0.0001) but BMI did not remain significantly correlated to presence of risk factors when controlling for WHR (p=0.09). These results indicate that abdominal distribution of body fat in women independently of general obesity is closely associated with metabolic risk factors including elevated blood pressure, a metabolic syndrome with increased risk for cardiovascular disease and non-insulin dependent diabetes mellitus. PMID- 16353585 TI - Do's and don't's for weight management: exercise is always good, but are some foods bad? PMID- 16353586 TI - What's in a name? Mr. Dickens' "Pickwickian" fat boy syndrome. PMID- 16353587 TI - A case of polysarka, in which death resulted from deficient arterialisation of the blood. 1866. PMID- 16353588 TI - Narcolepsy. 1889. PMID- 16353589 TI - Somnolence with cyanosis cured by massage. 1889. PMID- 16353590 TI - Case XXII. Excessive sleepiness. 1910. PMID- 16353591 TI - Extreme obesity associated with alveolar hypoventilation--a Pickwickian Syndrome. 1956. PMID- 16353592 TI - How can we do better? PMID- 16353593 TI - The influence of hydrocortisone (HC) on differentiation of adipose tissue is dependent on fetal age. AB - Elevated serum hydrocortisone (HC) levels are associated with larger fat cells and elevated levels of lipogenic and associated enzymes in late term pig fetuses from genetically obese dams. We have investigated the influence of HC status per se on these and other adipose tissue traits by chronically treating pig fetuses hypophysectomized (hypox) on day 70 with HC between either day 70 and 90 or 90 and 105 of gestation. Treatment with HC during both periods increased serum HC levels (P<.05) and increased fat cell size (P<.05) in the perirenal (PERI) and subcutaneous (SQ) depots, but failed to influence body weights, insulin-like growth factor-1 and insulin levels. Quantitative analysis of sections of PERI and SQ adipose tissue indicated that HC increased lipoprotein lipase (LPL), esterase and glucose-6-phosphate dehydrogenase (G6PDH) activities. The degree of esterase and G6PDH, but not LPL response to HC, was greater during the 90- to 105-day period than during the earlier period. HC significantly increased lipid accretion only in the SQ depot between 90 and 105 days. Overall, HC significantly augmented hypox-induced alterations in cellular and metabolic traits of developing adipose tissue. The general increase in fat cell size (21%) with moderate (SQ-105d) or no (PERI-90, 105d; SQ-90d) increase in lipid accretion indicates that HC either did not influence or decreased apparent fat cell number. Regardless, these data indicate that changes in serum HC per se may account for adipose tissue traits that characterize fetuses from genetically obese dams. PMID- 16353594 TI - Comparison of the lipolytic effects of norepinephrine and BRL 37344 in rat brown and white adipocytes. AB - The lipolytic effects of norepinephrine (a non-selective beta-agonist) and BRL 37344 (a selective beta3-agonist) were compared in isolated rat brown and white adipocytes. Norepinephrine and BRL 37344 maximally stimulated lipolysis in brown and white adipocytes, approximately 10 times above basal values. However, adipocyte sensitivity for BRL 37344 was greater than that for norepinephrine, particularly in brown adipocytes [the EC50 values (nM) for BRL 37344 and norepinephrine were 5 +/- 1 and 103 +/- 31 in brown adipocytes (P < 0.01) versus 56 +/- 9 and 124 +/- 17 in white adipocytes (P < 0.05), respectively]. On the other hand, the lipolytic effects of norepinephrine were totally blocked by 20-40 times superior concentrations of propranolol or bupranolol in brown as well as in white adipocytes. In contrast, the lipolytic effects of BRL 37344 were fully inhibited by concentrations of propranolol or bupranolol that were 200-1000 superior to the beta3 agonist concentration. The results demonstrate that: (1) the beta3-agonist BRL 37344 is as effective as norepinephrine for maximally stimulating lipolysis in rat brown and white adipocytes, (2) both adipocyte types are more sensitive to the lipolytic effects of BRL 37344 than to those of norepinephrine, (3) although bupranolol is a better antagonist than propranolol on BRL 37344-stimulated lipolysis, it cannot be considered as a specific beta3 antagonist, (4) brown adipocytes are 10 times more sensitive than white adipocytes to the lipolytic effects of BRL 37344, suggesting an important role of beta3-receptors in brown adipose tissue. PMID- 16353595 TI - Relation between BglII polymorphism in 3beta-hydroxysteroid dehydrogenase gene and adipose tissue distribution in humans. AB - The aim of this study was to investigate the association between a restriction fragment length polymorphism (RFLP) at the 3beta-hydroxysteroid dehydrogenase locus and adipose tissue distribution phenotypes. A total of 132 unrelated individuals from the Quebec Family Study were followed prospectively for an average period of 11.3 years. The BglII polymorphism in exon 4 of the 3beta-HSD gene was detected by PCR. Body mass, body fat, and regional fat distribution indicators were adjusted for age and age2 within each gender. Associations were assessed in unrelated adults with ANOVA across three genotypes. No association was found for the indicators of body mass, body fat, and regional distribution of adipose tissue measured in 1992. In women, the changes (difference between data collected in 1992 and at entry) in the sum of six skinfolds (p=0.04), abdominal skinfold (p=0.01), and abdominal skinfold adjusted (p=0.03) for the sum of six skinfolds at entry were related to the BglII polymorphism at the 3beta-HSD locus. These relations were not found in men, but they gained less body mass and body fat over the 11.3-year period. This suggests that sequence variation at the 3beta HSD locus or in neighboring genes on chromosome 1 may contribute to individual differences in body fat content and adipose tissue distribution in adult women, particularly in abdominal adipose tissue deposition as they grow older and gain body fat. PMID- 16353597 TI - LY226936 administered orally and centrally to obese Zucker rats suppresses food intake and body weight gain. AB - LY226936, methylcarbamothoic acid-S-(4,5-dihydro-2-thiazolyl) ester, is a new compound that, when administered to obese Zucker rats, caused reduced food intake. LY226936 reduced the food consumption after a single oral dose of 50 and 100 mg/kg. On chronic oral administration to meal-fed obese (5 to 35 mg/kg. once daily) and to fed obese and lean (15 mg/kg. twice daily) Zucker rats, LY226936 reduced food intake and body weight gain for periods ranging from 40 to 48 days. The effect on both parameters was statistically significant. There is no evidence in our studies that tolerance to the actions of LY226936 developed. LY226936 decreased the consumption of both high carbohydrate and high fat diets. Food consumption of meal-fed obese Zucker rats was reduced significantly each time a single dose of 10 ugm LY226936 per rat was infused intracerebroventricularly. None of the receptors studied (mu and kappa opioid, CCK, serotonin, neuropeptide Y, galinin, N-methyl-D-aspartic acid) appeared to bind LY226936 and therefore, appear not to be involved in the depression of food intake by the obese Zucker rat. PMID- 16353596 TI - Development and validation of computed tomography derived anthropometric regression equations for estimating abdominal adipose tissue distribution. AB - The purpose of this study was twofold: (1) to develop multiple regression equations for predicting computed tomography (CT) derived intra-abdominal (IAF), subcutaneous (SCF), and total (TOTF= IAF+SCF) abdominal adipose tissue areas from anthropometric measures in adult white males with a large range of age (18-71 years) and percent body fat (2.0-40.6); and (2) to validate the new and existing equations that used similar Hounsfield Units (HU) for determining IAF for estimating these fat depots. One hundred fifty-one white male subjects had IAF, SCF, and TOTF determined by a single CT scan, skinfold and circumference measures taken and body density determined. Linear intra-correlations and factor analysis procedures were used to identify variables for inclusion in stepwise multiple regression solutions. IAF was estimated from age, waist circumference, the sum of mid-thigh and lower thigh circumferences, and vertical abdominal skinfold. SCF was estimated from age, umbilicus circumference, chest and suprailiac skinfolds. TOTF was estimated from age, body mass index (BMI), chest skinfold, and umbilicus circumference. R2 for IAF, SCF, and TOTF was .73, .77, and .86 respectively. The existing and the new equations were validated on an independent sub-sample of 51 subjects. The only existing equation that met validation criteria had a validation R2 = .67 for IAF. All three new equations met validation criteria with R2 validations of .75, .79, and .85 for IAF, SCF, and TOTF respectively. It is concluded that the new equations might be used as an inexpensive estimation of IAF, SCF, and TOTF in adult white males varying greatly in age and percent body fat. PMID- 16353598 TI - Genetic factors influence the response of energy metabolism, body weight, and body composition to exercise. PMID- 16353599 TI - Antiobesity and antidiabetic beta-agonists: lessons learned and questions to be answered. AB - In several species of obese animals, a group of phenethanolamine beta-agonists stimulates lipolysis and thermogenesis, resulting in the loss of body fat and weight. Brown adipose tissue is considered to be the major target tissue for the antiobesity activity of these compounds. Independent of this antiobesity activity, some of these compounds are also antidiabetic and increase muscle mass. Based on the pharmacological profile of these compounds, a beta3-receptor was proposed and characterized in mouse, rat, and humans. The beta3-receptor in brown adipose tissue has been suggested to mediate the antiobesity activity of these beta-agonists. Whether this receptor is responsible for the antidiabetic activity and whether there is a linkage between the antiobesity/antidiabetic activity and the nutrient partitioning activity is not clear. Clinical trials with these mixed beta-agonists showed marginal antiobesity effects when caloric intake of subjects was restricted. Insulin sensitivity was also improved in some of the trials designed to test the antidiabetic activity of these compounds. Side effects included tachycardia and tremor. To eliminate these side effects, a second generation of compounds was selected for its agonist activity on rat beta3 receptors. Clinical trials with these compounds have shown little increase of energy expenditure even at high doses. Successful development of an antiobesity and antidiabetic drug from this class of compounds will require the elucidation of the physiological role of the human beta3-receptor and the regulatory mechanism between fuel efficiency and feeding behavior. PMID- 16353600 TI - Harvey Cushing and the neuroendocrinology of obesity. PMID- 16353601 TI - The basophil adenomas of the pituitary body and their clinical manifestations (pituitary basophilism). 1932. PMID- 16353602 TI - Is DL-fenfluramine a potentially helpful drug therapy in overweight adolescent subjects? AB - We have studied the therapeutic effects of two different doses (30 mg and 60 mg, twice daily) of DL-fenfluramine (DL-F) in, respectively, prepuberal (11-13 years old) and adolescent subjects (14-17 years old). Sixty-eight obese subjects were recruited for this study (22 boys, 36 girls, aged 10-17 years old) with body mass index ranging from 24.5 to 44.0 kg/m2, absolute weight ranging from 37.0 to 119.5 kg and % over IBW ranging from 122% to 260%. Results were compared to a placebo treated group of obese adolescent patients (n = 17), 6 boys and 11 girls, aged 10 17 years old, BMI ranging from 26-44 kg/m2, absolute weight 53.1 to 96.5 kg, and with 129% to 253% over IBW. In the DL-F-treated subjects most patients (n = 41) had a continuous weight loss during 12 months but 27 individuals were unable to lose any additional weight after the initial 6 months of the trial. Taken together 65% of all patients lost weight during DL-F treatment (12 months) whereas only 17.4% of the placebo group lost a significant (> 10% BMI) amount of excess weight. Also the placebo group had a higher withdrawal rate (57%) as compared with the DL-F-treated group (24%). There was a significant (p < 0.05) decrease of the mean +/- SD of the BMI (at 6 and 12 months of therapy). No significant change of the BMI was observed for control group. Minor adverse side effects consisted of a brief period of drowsiness and dry mouth. Our findings indicated that the continuous administration of DL-Fenfluramine might help obese adolescent subjects adhere to a diet and to maintain the weight loss achieved without major or harmful adverse effects. PMID- 16353603 TI - A randomized double-blind crossover study of the antiobesity effects of etiocholanedione. AB - Etiocholanedione (ED), a natural metabolite of dehydroepiandrosterone, has antiobesity effects in animals when given orally and is nontoxic. We carried out a trial of oral ED in obese humans. In a 20-week randomized double-blind crossover study, 14 subjects lost significantly more weight and body fat during treatment with oral ED, 4 gm daily, than during placebo administration. Mean weight loss during ED administration was 2.8 +/- 5.5 kilograms, which was equivalent to 0.53 +/- 0.91 kilograms per week per 100 kilograms of body fat; mean weight change during placebo administration was essentially zero: +0.21 +/- 4.2 kg, or +0.04 +/- 0.74 kg/wk/100 kg body fat. The difference between the weight changes in the two periods was significant: for delta kg, P < 0.05; for delta kg/wk/100 kg body fat, P < 0.03. Densitometric measurement of body fat content showed that the mean weight loss coincided almost exactly with the mean decrease in fat content; thus, over the 10-week period of ED administration, the mean fat loss was about 5% of the initial body fat content. Three of the obese subjects had strikingly greater fat loss, about 18%, 19%, and 25% of the initial body fat content. There were no significant subjective or objective side effects of ED administration. PMID- 16353604 TI - Do obese eat faster than lean subjects? Food intake studies in Pima Indian men. AB - Food intake rate has previously been derived from observation of eating behavior in laboratory settings or in public eating establishments. Although it has been suggested that obese individuals eat faster than lean individuals, observations of such an "obese eating style" have yielded mixed results. In the present study, the relationship between ad-libitum food intake rate and obesity was evaluated over 4 days on a metabolic ward in 28 healthy Pima Indian men (Mean +/- SD; 29 +/ 7 y, 100.4 +/- 27.1 kg, 33 +/- 10% body fat) using an automated food selection system containing a large variety of foods. Total energy intake averaged 18829 +/ 3299 kJ/d consisting of 47 +/- 4, 40 +/- 3, and 13 +/- 1 percent of carbohydrate, fat and protein, respectively. The average meal duration was 25 +/- 7 min. Food intake rate was 68 +/- 21 g/min while carbohydrate, fat and protein intake rates were 23 +/- 6, 9 +/- 3 and 6 +/- 2 g/min, respectively. Food intake rate correlated negatively with % body fat (r = -0.61, P < 0.01). Similar relationships were found between the intake rates of carbohydrate, fat and protein and body fatness. Only prospective studies will indicate whether a slow food intake rate may contribute to the etiology of obesity by possibly reducing satiety. PMID- 16353605 TI - The social self, body dissatisfaction, and binge eating in obese females. AB - We examined the relationship of the social self to body dissatisfaction and to binge eating in two clinical samples of obese females. In the first study, with 32 nonbingeing obese females, social self measures of Public Self-Consciousness and Social Anxiety were positively correlated with body dissatisfaction. Self esteem was negatively correlated with body dissatisfaction. In the second study, 11 obese binge eaters were compared with 11 matched nonbingeing obese females. Binge eaters were characterized by significantly higher levels of social anxiety and body dissatisfaction than the obese nonbingeing controls. These findings support the hypothesized links of social self concerns to body dissatisfaction and to binge eating in obese females. PMID- 16353606 TI - Racial differences in selection of ideal body size by adolescent females. AB - Cultural differences may partially account for the fact that more black women than white women are overweight in the United States. This study measured perceptions of ideal body size among 93 black and 80 white females, 14-17 years old, who were randomly selected from three public high schools in a southeastern state. The subjects' height and weight were measured along with their estimates of their mother's body size, weight control attitudes, and demographic variables. Blacks preferred a significantly larger body size than whites when asked to select ideal body size (p = 0.045). Subjects who estimated their mother's size to be larger, also selected a larger ideal body size (p = .047). Those who perceived themselves as too fat were more likely to skip meals to help control their weight, compared with those who perceived themselves as normal or too thin (p = .003). Approximately 30% of the sample was overweight or obese. There were not significant racial differences in weight or BMI. These results suggest that overweight is more acceptable among black females than among white females and may help explain why more black females are obese. PMID- 16353607 TI - Determinants of free-living energy expenditure in normal weight and obese women measured by doubly labeled water. AB - Total free-living energy expenditure (TEE) was measured in 9 normal weight controls and 5 obese women using the doubly labeled water (DLW) method. Resting energy expenditure (REE) and the thermic effect of food (TEF) were measured by indirect calorimetry and the energy cost of physical activity (PA) calculated by deduction, in order to quantify the components and identify determinants of free living TEE. Although REE was quantitatively the major component of TEE in both groups, PA best explained the variability, contributing 76% to the variance in free-living TEE. The obese women had elevated values for TEE (12397 +/- 2565 vs. 8339 +/- 1787 kJ/d, mean +/- SD; p < 0.005), compared with the control women. PA (5071 +/- 2385 vs. 2552 +/- 1452; p < 0.05) and REE (6393 +/- 678 vs. 5084 +/- 259; p < 0.0005) were also raised in the obese, whereas TEF was not significantly different between the groups, accounting for 7.6% of energy expenditure for the obese and 8% for the control subjects. Body weight was the single best determinant of mean daily free-living TEE across both groups. We conclude that PA and body weight are the main determinants of free-living TEE. PMID- 16353608 TI - Pharmacotherapy for childhood obesity? Maybe for some. PMID- 16353609 TI - A perspective on the hormonal abnormalities of obesity: are they cause or effect? AB - Studies in our laboratory and elsewhere have demonstrated numerous abnormalities of steroid and polypeptide hormone secretion in obesity: hyperestrogenemia and hypogonadotropic hypogonadism in obese men; diminished SHBG levels in both sexes; elevated free testosterone and free estradiol in obese women; PCOS-like gonadotropin and sex-hormone abnormalities in obese women; elevated serum insulin in both sexes; blunted stimulability of prolactin, growth hormone, and vasopressin in both sexes; and elevated basal levels and blunted stimulability and suppressibility of beta-endorphin in both sexes. All of these abnormalities have been clearly shown to be partly or completely reversible with weight loss, with the exception of the endorphin abnormalities. In that area, four out of the five studies reported show no reversibility with weight loss. Reversibility of nearly all the hormonal abnormalities of obesity (i.e., all but the hyperendorphinemia) by weight loss suggests that none of them is causative of obesity. Nevertheless, some of the reversible abnormalities may secondarily amplify the morbidity associated with obesity: the hyperinsulinemia may be related to the increased risk of hypertension, hyperlipidemia, coronary disease, and Type II diabetes; the elevated levels of free estradiol in obese women may be related to their increased risk of breast and endometrial cancer. The role of hyperendorphinemia in obesity clearly requires further investigation, since it is the only observed hormonal abnormality that appears to be non-reversible by weight loss, and also since there seems to be increased sensitivity to beta endorphin in obesity. The possibility that endorphin abnormalities may be causal in obesity cannot be ruled out. PMID- 16353610 TI - Commentary on classics in obesity. 1. Quetelet: quantitative medicine. PMID- 16353611 TI - A treatise on man and the development of his faculties. 1842. PMID- 16353612 TI - Higher calorie content preserves myocardial electrical activity during very-low calorie dieting. AB - We investigated the effect of calorie and protein content of very-low-calorie diets for weight loss upon myocardial electrical activity. Patients were followed on very low calorie diets for up to 20 weeks. Study No. 1 had 16 obese patients (4 men and 12 women). Study No. 2 had 113 obese women. Both study groups were consecutive samples with patients who had BMI > 30. In Study No. 1 patients were given 400 kcal/d with 50 gm of protein and 45 gm of carbohydrates over 12 weeks. In Study No. 2 patients were given either Metabolic Nutrition Program (MNP) which contained 600 kcal/d with 70 gm protein and 70 gm carbohydrates, or Optifast-70 which contained 420 kcal/d with 70 gm protein and 32 gm carbohydrates, for up to 20 weeks. Electrocardiograms were obtained at baseline and during the third month in Study No. 1, and monthly in Study No. 2. The sum of the voltage in leads I+II+III and the length of the QT interval were derived from the electrocardiograms. In Study No. 1 the QRS voltage decreased from 2.77 +/- .18 mv to 2.45 +/- .17 mv (p < .02). Patients lost 18.1 kg +/- 4 kg and the QT interval remained unchanged. In Study No. 2 at 20 weeks the QRS voltage decreased on Optifast-70 (p < .001) in comparison to MNP, in which the QRS voltage remained unchanged. Patients on Optifast-70 lost 17.5 +/- .53 kg, patients on MNP lost 18.5 +/- .66 kg and the QT interval in both groups remained unchanged. The formula diets tested of less than 600 kcal/d were associated with a decrease in electrocardiographic voltage. PMID- 16353613 TI - Black and white females' perceptions of ideal body size and social norms. AB - Different cultural norms and standards for appropriate female body size might contribute to the disparity in obesity rates between black and white adult females (46.0% and 24.6% respectively). The purpose of this study was to measure adolescents' perceptions of ideal size and social norms regarding female body size as well as adolescents' perceptions of significant others' evaluation and expectations of the adolescents' body size. Subjects included 437 adolescent girls (247 white and 190 black) aged 13 to 19 (x = 14.9, SD = .979) from six randomly selected public schools. The subjects, heights and weights were measured. Responses to a body image questionnaire and a series of nine female body drawings (arranged ordinally, 1 to 9, from thinnest to heaviest) were analyzed using the General Linear Model and Logistic Regression. The female body size considered ideal by black females was significantly larger than the size selected as ideal by white females (x = 3.47 and x = 3.13 respectively, p < 0.001). Black females were two times more likely than white females to describe themselves as thinner than other girls their age (O.R. = 2.01, 95% C.I. 1.34, 3.01) and seven times as likely to say that they were not overweight (O.R. = 7.08, 95% C.I. 3.72, 13.45). White females wanted to be a smaller size than they currently were and felt encouraged by significant others to lose weight or reduce their size. Black females did not indicate as great a desire as whites to be smaller and they tended to feel that their size was considered satisfactory by significant others. Only subjects from the low SES group perceived that significant others wanted them to gain weight. The differences between black and white subjects' beliefs and perceptions about body size norms may explain, in part, why heavier body weights persist in some cultural groups. PMID- 16353614 TI - Body size perceptions and eating attitudes in elderly men. AB - The majority of studies on eating attitudes, dieting and body size perceptions have focused on young adults and women. This study examined these attitudes in 334 black and white men, ages 55 to 98 years, who were members of the Charleston Heart Study cohort. Associations of the eating attitude variable with race, education and weight status were examined. Eighty-two percent of the overweight white men studied had dieted to lose weight, whereas only 49% of slimmer white men had dieted. In contrast, overweight black men did not diet more than slimmer black men. Overall black men dieted less than white men (37% reported dieting). Black men who were high school graduates were 1.3 times more likely to have dieted than were less educated black men. Overweight white men were over twice as likely as slimmer white men to feel guilty after overeating. This difference was not found in overweight versus slimmer black men. Education was not associated with measured body mass index (BMI) or perceived or ideal body size. However, there were some racial differences in these variables. White men preferred to be slightly thinner than black men (ideal BMI 25.6 vs. 26.1 kg/m2), and the difference between actual and desired BMI was 0.6 kg/m2 greater in white than in black men. These results indicate that effects of race and weight status on eating restraint and body size perceptions seen in younger subjects and in women are also present, at least to some degree, in elderly men. PMID- 16353615 TI - The relationship between daytime and nighttime food intake in an obese night eater. AB - Some obese individuals consume food during awakenings from nighttime sleep. Three studies were conducted on a 28-year-old morbidly obese male with chronic sleeping complaints and insignificant weight loss, despite self-reported daily caloric restriction: I. For 3 mo, the subject recorded food intake for 24-h periods. Mean daytime intake was 1286 kcal +/- 386 (SD), and mean nighttime intake was 1036 kcal +/- 487 (SD). Caloric values of daytime and nighttime intake were negatively correlated, r = -0.22, df= 82, p < .05. II. Seven consecutive 24-h food intake recordings were obtained with an automated formula dispenser when the subject was an inpatient on a metabolic ward and received ad libitum formula as his sole food source. Mean daytime intake was 1245 +/- 662 (SD), and mean nighttime intake was 231 +/- 236 (SD). There was a non-significant negative correlation between daytime and nighttime intake, r = -0.32, df = 5, NS. III. The subject underwent polysomnographic studies on 2 non-consecutive nights, following the administration of either a low (600 kcal) or high (1800 kcal) daytime caloric condition. The subject, upon awakening from nighttime sleep, could eat from a platter of sandwich quarters placed at his bedside. The addition of 1200 kcal to daytime intake decreased nighttime intake by 654 kcal, or by 55% of the additional calories delivered during the day. The three studies (I, II, and III) show that daytime food intake can be negatively correlated with nighttime intake, and that daytime intake can influence nighttime intake in a documented obese night-eater. PMID- 16353616 TI - No specific effect of fluoxetine treatment on fasting glucose, insulin, lipid levels, and blood pressure in healthy men with abdominal obesity. AB - In this paper we investigated the effect of fluoxetine (60 mg/d) on serum lipids, glucose and insulin concentrations and blood pressure by means of a randomized, double-blind placebo controlled trial. Thirty-eight overweight (BMI: 26-30 kg/m2), nondiabetic, nonhypertensive men with an abdominal fat distribution (waist/hip ratio: > 0.97) received dietary advice and placebo or fluoxetine for 12 weeks. The changes in serum parameters and blood pressure in the fluoxetine treated group were not different from the placebo treated group, despite a significantly larger weight loss in the fluoxetine group. In both groups serum total-cholesterol concentrations, serum LDL-cholesterol concentrations and the HDL/LDL ratio were significantly improved after treatment. Reductions in fasting glucose concentration and systolic blood pressure were only significant in the placebo group. A reduction of serum triglycerides and an increase of HDL cholesterol were found in the fluoxetine treated group. In the total study population the changes in serum lipids seemed to be more strongly related to the change in total body fat or subcutaneous abdominal fat (assessed by MRI) compared to the change in visceral fat. The improvement of most of the serum lipids was related to the change in total body fat independent of the mechanism for attaining this fat loss. Our results indicate that fluoxetine treatment has no specific effect beyond that expected for weight loss on serum lipid, glucose and insulin concentrations, and blood pressure in overweight men. PMID- 16353617 TI - Needed for NAASO: A code of ethics. PMID- 16353618 TI - Obesity in minority populations: an epidemiologic assessment. AB - This article reviews issues relating to the prevalence, health implications, and prevention and treatment perspectives of obesity in U.S racial and ethnic minority groups. The growing interest in obesity in minority populations reflects an awareness of the high prevalence of obesity among black, Hispanic, Asian and Pacific Islander and Native Americans as well as a generally increased interest in minority health. In addition, the fact that some aspects of obesity among minorities differ from those in whites suggests that new insights may be gained from studying obesity in diverse populations. However, there are many methodological problems to be overcome, including some that arise from the way minority groups are defined. Under the assumption that all obesity results from a period of sustained positive energy balance at the individual level, an epidemiologic explanation for the excess of obesity in minorities at the population level seems readily apparent. A surplus of obesity-promoting forces and a deficit of obesity-inhibiting forces, caused by secular changes in food availability and physical activity, accompany the early phases of modernization and economic advancement. The high prevalence of obesity in minority populations can be viewed as a function of the slope and timing of these secular changes. Genetic predisposition, cultural attitudes, and exposure to maternal obesity and diabetes in utero may be potentiating factors. In this context, interventions targeting individuals would seem inevitably to put racial and ethnic minority groups on the path toward the same weight control crisis now observed in the majority white population. This suggests that the underlying causes of the societal energy balance problem must be addressed at the population level in order for effective clinical approaches to be developed for minority populations with a high obesity prevalence. PMID- 16353619 TI - Lavoisier and scientific revolution: the oxygen theory displaces air, fire, earth, and water. PMID- 16353620 TI - Memoir on heat. Read to the Royal Academy of Sciences, 28 june 1783. 1783. PMID- 16353621 TI - [Biological effects of microwave radiation of low nonthermal intensity (regarding the maximal admissible values)]. AB - There is still much controversy in scientific approaches to setting limits of exposure to radio-frequency electromagnetic radiation in Russia, Europe and USA. The stumbling stone is recognition or denial of potential risk of biological effects from nonthermal electromagnetic radiation (to 1 mW/cm2). The authors review results of experimental investigations performed in Russia over the years since 1960s to study the direct and delayed effects of ultra-high and microwave frequencies of low non-thermal intensity EMR, and delayed negative effects in employees based on the data of several follow-up epidemiological studies. This evidence was used as a baseline for establishing the Russian limits for chronic exposure to, thresholds of harmful effects and the maximal admissible values of EMR within the above frequencies. PMID- 16353622 TI - [Concept of the telemedicine system for a Martian mission]. AB - The telemedicine concept for a mission to Mars is presented including the basic operational units and subsystems, and problems of elaboration and creation of the telemedicine system with the use of multilevel structure functional modeling. PMID- 16353623 TI - [Shifts in human venous hemodynamics in long-term space flight]. AB - Veins and venous hemodynamics before, during and after long-term space flights (SF) were evaluated with the ultrasonic (B-scan, Doppler) techniques and occlusion plethysmography. Fifteen cosmonauts were investigated in eight SFs of about 6 months in duration. In three SFs, 30 investigations of the main veins in different parts of the body were fulfilled with participation of 7 cosmonauts. In five SFs, 48 investigations of venous distensibility in legs were fulfilled with occlusion plethysmography with participation of 8 cosmonauts. Femoral veins tended to increase distension throughout SF. Exaggerated leg veins distensibility and compliance was persistently observed during application of occlusion from the early period of SF (wks 1-2) which, in our opinion, extenuates the reduction in LBNP tolerance and post-flight degradation of orthostatic stability. These shifts were growing markedly till months 2-3 of exposure to microgravity and tended to stabilize at the worst (as compared with prelaunch values) level at the end of six-month SFs. PMID- 16353624 TI - [Evaluation of +Gz tolerance following simulation of 8-hr flight]. AB - Tolerance of +Gz (head-pelvis) centrifugation of pilots was evaluated following simulation of a long flight on single-seat fighter. The experiment involved 5 test-subjects who were exposed to +Gz before and after simulated 8-hr flight with a growth gradient of 0.1 u/s without anti-g suits and muscles relaxed; in addition, limiting tolerance of intricate profile +Gz loads of 2.0 to 9.0 units with a growth gradient of 1.0 u/s of test-subjects in anti-g suits (AGS) with a change-over pressure valve in the peak mode using muscle straining and breathing maneuvers. To counteract the negative effects of extended flight, various seat configurations: with a back inclination at 30 degrees to the +Gz vector and changeable geometry with a back inclination at 55 degrees to the vector. The other counter-measures applied were cool air shower, suit ventilation, physical exercises, lower body massage with AGS, electrostimulation of the back and lumber region, profiling of the supporting and soft parts of the seat, and 30-s exposure to +5 Gz. Hemodynamic and respiration parameters as well as body temperature were measured in the course of 8 hrs of flight and during and shortly after centrifugation. According to the results of the investigation, seat inclination at 55 degrees to the +Gz vector and tested system of countermeasures prevent degradation of tolerance of large (9 u.) loads following 8-hr flight simulation with the use of the modern anti-g gear, PMID- 16353625 TI - [Characteristics of cosmonauts' metabolism after extended missions on the international space station]. AB - In eight Russian members of the ISS crews 1 to 5 enzymes and some other metabolites were analyzed in blood in the pre- launch period, on the landing day, and on days 6-8 and 14-19 of recovery. Deviations in the biochemical parameters were typically within the physiological norm without clinical implications. Biochemical deviations on the landing day seemed to relate to metabolic shifts due to long-term microgravity and, on the other hand, were a reaction to acute gravitational stress on return to 1 g. Dynamics of a number of parameters was markedly different as compared with the data from cosmonauts who had made long missions to the Mir space station, but this can be attributed to different g profiles during descent and blood sampling schedule. In two to three wks the parameters regained normal values suggesting that negative effects of readaptation to the Earth's gravity were fully remedied. PMID- 16353626 TI - [Impedancimetry of the hydration status and body composition of head-down tilted males]. AB - Studies of the hydration status and body composition of 6 normal males was performed before and after 7-d head-down tilt (-8 degrees) with the use of a portable impedance-measuring device. Fluid sectors in the organism tended to proportionally reduce. By the end of the experiment, the total body fluid was found reduced by 6.4 = 0.9%; the intracellular and extracellular volumes fluids were reduced by 6.4 +/- 0.9% and 6.5 +/- 0.9 %, respectively. Body mass changed but little, and not to the same extent in each subject. Lean body mass measured with the device showed a small reduction (2.0 +/- 0.6 %), whereas the fatty body mass grew in all subjects by 14.0 +/- 3.1% on the average. These results lead to the conclusion that during the 7d head-down tilt the hydration status of the human body grew low concurrent with loss in muscular mass and increase in the body fatty mass. PMID- 16353627 TI - [Investigation of the anti-motion sickness effect of 3-hydroxypyridine derivatives]. AB - Experiments with rats showed that three out of 12 3-hydroxypyridine derivatives (ethyl-methyl hyd- roxypyrine succinate, SK-132 and IBCP-2 - had an anti-motion sickness effect stronger than of scopolamine, the reference vestiboloprotector. The anti-motion sickness effect of ethyl-methyl hydroxypyrine was also demonstrated in experiments with cats. Apparent anti-motion sickness effect of ethyl-methyl hydroxypyrine (mexydol) was found in 69% of healthy male volunteers which is comparable with the effect of scopolamine (62%). In experiments with immobilized cats (myorelaxation drugs) the microelectrode technique and microontoiphoresis of physiologically active substances revealed that ethylmethyl hydroxypyrine influences the majority of neurons in the medial vestibular nucleus (61%). Suppression of cell spontaneous activities in more than one half of cases can be stopped completely or attenuated significantly by bicucculine, a specific GABA(A)-receptor antagonist. In 42% of neurons ethyl-methyl hydroxypyrine subdues the response to vestibular stimulation which is likely to underlie the anti motion sickness effect. PMID- 16353628 TI - [Disorders in rat's higher nervous activity in the course of chronic gamma irradiation]. AB - Investigations of early disorders in rat's higher nervous activity in the course of chronic gamma-irradiation (elaboration of the conditioned defensive reflex of avoidance "under the ray") showed that already in 10-15 minutes since the beginning of exposure (total dose of approx. 0.75-1.15 Gy) the conditioned reflex parameters degraded significantly with a general trend toward an increased excitation and disruption of active inhibition in the higher sections of the central nervous system (CNS). The most dramatic disorders occurred in the middle of the period of exposure (total dose of approx. 4-5 Gy). There were some individual differences in the CNS functional reaction to irradiation. PMID- 16353629 TI - [Investigation of the main gas-exchange parameters in the Mongolian gerbil in view of a space experiment]. AB - A pressurized module with a self-sustained life support system is being designed for spaceflight experiments with Mongolian gerbils (Meriones unguiculatis). As a rule, these rodents do not drink free water taking minimum of water available in nourishment Although the gerbil is a common test object, it has never been flown in space. Specifically, nothing is known about the animal gas-exchange parameters critical for the development of life support systems. Our purpose was to shed light on the gerbil's gas-exchange demands and thereupon to prototype the air supply system in simulation experiments. On the average, a group of (n = 9, mean body mass of 45 g) consumed 0.044 ml/g/min of oxygen and produced 0.024 ml/g/min of CO2. The proposed design of the animal gas supply system controls oxygen and carbon dioxide at the atmospheric concentrations. Data of these experiments were used to write specifications for animals maintenance module KONRTUR to be launched aboard space vehicle FOTON-M. PMID- 16353630 TI - [Octafluoropropane inhalation toxicity]. AB - Wistar rats were used to study inhalation toxicity of octafluoropropane (OFP, freon-218) at the following concentration: 300 g/m3 (4-hrs), 30 g/m3 (0.5 to 4 hrs), 3 g/m3 (8 hrs), and 0.3 g/m3 (16 hrs). According to the histological analysis, OFP at the concentrations of 300 and 30 g/m3 had a politrophic toxic effect. Target organs were the lung, trachea, bronchus, heart, kidney, and the adrenaL There were dystrophic and necrobiotic lesions in the upper airways epithelium. Subacute and chronic vesicular bronchiolitis developed on days 7 and 14, respectively. Visceral organs and brain were found plethoric and the lung was found hemorrhagic. Similar lesions were seen in the trachea, bronchus, lung, liver, spleen, kidney, adrenal, heart, and the brain. Lipid redistribution was observed in the adrenal cortex and vascular reactions of renal tissue with juxtamedullar blood shunting. Toxicity of small OFP concentrations (3 and 0.3 g/m3) was distinguished by an extended aftereffect, these concentrations did not cause visible pathomorphologic changes but gave rise to an extended pathologic process detectable by biochemistry. In all concentrations, OFP impacted erythrocyte metabolism changing the lipid composition of cell membrane and activating membrane-bound adenosinetriphos-phatases. The activities of hepatocyte and myocardiac cytoplasmatic enzymes were altered in blood plasma. Increased malonic dialdehyde in blood plasma and decreased cell antioxidant GSH in erythrocytes suggested exaggerated lipid peroxidation. These data point to the necessity of revising the existing limits for OFP concentrations in air of populated areas, working areas, and closed human environments. PMID- 16353631 TI - [Effects of medium with elevated partial pressure of argon and nitrogen on morphology of Escherichia coli colonies]. AB - The purpose was to study morphological specifics of microbial colonies cultivated in argon-containing gas medium. Investigated were computerized images of the colonies of museum strains Escherichia coil K-12. It was found that combination of elevated argon pressure (1 ata) and nitrogen pressure (1.8 ata) increased the maximal density of microbial colonies and decreased the colony size as compared with cultivation in argon only. PMID- 16353632 TI - Mitochondrial large ribosomal subunit sequences are homogeneous within isolates of Glomus (arbuscular mycorrhizal fungi, Glomeromycota). AB - Partial sequences of the mtLSU rDNA were obtained from the arbuscular mycorrhizal (AM) fungi Glomus proliferum (isolate DAOM 226389) and G. intraradices (isolates JJ291 and BEG75). The exon sequences of the two species showed regions of strong divergence. There was no evidence of intra-isolate sequence heterogeneity as it is found in variable regions of nuclear ribosomal genes of Glomeromycota. In G. intraradices JJ291, two introns were found in the partial LSU sequence. One of the introns contained an ORF for a putative site-specific homing endonuclease of the LAGLIDADG family. In G. intraradices BEG75, one of the introns was missing and the other had a DNA sequence distinct from JJ291. G. proliferum had no introns in the region sequenced. A PCR primer was designed to amplify the fragment of the mtLSU of a different, distinguishable G. intraradices genotype from colonized roots of a field sample. These mitochondrial gene sequences are the first reported from the phylum Glomeromycota. Our findings indicate that the intra-individual sequence heterogeneity of the Glomeromycota may be a peculiar feature of the nuclear genes. Therefore, mtLSU and its introns have the potential to be highly sensitive genetic markers for these fungi in the future. PMID- 16353633 TI - A complex of three new white-spored, sympatric, and host range limited Geosmithia species. AB - All hypocrealean species of the genus Geosmithia are anamorphic fungi with connections to bark beetles. G. fassatiae, G. langdonii and G. obscura are described as new sympatric species associated with Scolytus carpini, S. intricatus and S. rugulosus in Central Europe. The species represent a complex of three sister taxa with affinities to G. flava that may be distinguished by differences in morphology, unique RAPD patterns and by sequences of ITS region rDNA. Intraspecific variability and habitat specificity of new species is described and discussed. The high morphological, genetic and ecological uniformity suggest that these Geosmithia spp. are recently derived. A key to all accepted hypocrealean species of the genus is provided. PMID- 16353634 TI - Pythium phragmitis sp. nov., a new species close to P. arrhenomanes as a pathogen of common reed (Phragmites australis). AB - During a study on the occurrence and pathogenicity of oomycetes in the reed-belt (Phragmites australis) of Lake Constance (Germany), a new Pythium resembling the important cereal pathogen species complex P. arrhenomanes/P. graminicola was consistently isolated from necrotic mature reed leaves and reed rhizosphere samples. The new species proved to be significantly more aggressive towards reed leaves and seedlings in vitro than related species. It is characterised by filamentous, inflated sporangia and plerotic oospores with usually more than one antheridium. ITS and cox II sequence data indicate this new species shares a common ancestor with P. arrhenomanes, but the sequence differences are clearly consistent with a divergence of the two taxa and with P. phragmitis being a distinct species. ITS 1 and 2 of 15 isolates of the taxon consistently differed from P. arrhenomanes by 13 positions. Sequence analyses of the cox II gene confirmed the new species' phylogenetic position. This paper gives a formal description of the taxon as P. phragmitis sp. nov., providing information on morphology, ecology and pathogenicity in comparison to related species. As indicated by the close association to Phragmites australis, the high aggressiveness towards reed leaves and seedlings, and the abundance in the investigated stands, Pythium phragmitis might act as a reed pathogen of considerable importance, in particular under flooding situations. PMID- 16353635 TI - Botryosphaeria species from Eucalyptus in Australia are pleoanamorphic, producing dichomera synanamorphs in culture. AB - Species within the genus Botryosphaeria include some of the most widespread and important pathogens of woody plants, and have been the focus of numerous taxonomic studies in recent years. It is currently accepted that anamorphs of Botryosphaeria belong to two distinct genera, Fusicoccum and Diplodia. Species within the genus Fusicoccum commonly produce aseptate, hyaline conidia. In the present study, fungi were isolated from foliage and wood of Eucalyptus in native forests and plantations in Australia. Although these fungi produced Dichomera anamorphs in culture, they clustered within the Fusicoccum clade of Botryosphaeria based on their ITS sequence data. Four species, Botryosphaeria dothidea, B. parva, B. ribis and B. australis produced Dichomera conidia in culture. The Dichomera synanamorphs are described for these four species of Botryosphaeria. In addition, falling within the Fusicoccum clade of Botryosphaeria, two species were found to be distinct from previously described Botryosphaeria spp. based on their ITS sequences, but synonymous with D. versiformis and D. eucalypti. These observations are currently unique to isolates from host trees within the genus Eucalyptus in Australia, and the pleoanamorphic nature of these species is discussed. PMID- 16353636 TI - Genetic diversity and population structure among strains of the entomopathogenic fungus, Beauveria bassiana, as revealed by inter-simple sequence repeats (ISSR). AB - Inter-simple sequence repeat (ISSR) markers were used to investigate genetic diversity among 39 isolates of Beauveria spp. (36 B. bassiana, one B. brongniartii, one B. amorpha, and one B. velata) isolated from different insect hosts and geographical origins. Eighteen among 33 primers that contained different simple sequence repeats (SSR) were chosen for their reproducibility and high polymorphism. Considerable intraspecific and interspecific variability among the isolates of Beauveria spp. was revealed. One hundred sixty-eight highly reproducible fragments were amplified in all 39 isolates with an average of 9.3 markers per primer; among these, 161 (95.8%) were polymorphic. For 36 B. bassiana isolates, 8.9 (1-13) markers per primer were scored, and a total of 123 fragments were amplified, in which 102 (82.9%) were polymorphic. Among the 168 polymorphic bands, 7 bands were considered to be specific for B. brongniartii isolate Bbr06, 14 bands for B. amorpha isolate Ba08, and 18 bands for B. velata isolate Bv0. Within 36 B. bassiana isolates, genetic similarity ranged from 0.651 to 0.972. However, the genetic similarity values among different Beauveria species ranged from 0.411 to 0.720, suggesting that ISSR technique was successful in differentiating the three closely related species from B. bassiana. The results also indicated that there was a certain association between B. bassiana isolates and their geographical origins, but no clear correlation between those isolates and their insect hosts. The present study suggested that ISSR markers can be used as robust molecular markers for the population genetics, epidemiological and ecological studies of entomopathogenic fungi. PMID- 16353637 TI - NEP1 orthologs encoding necrosis and ethylene inducing proteins exist as a multigene family in Phytophthora megakarya, causal agent of black pod disease on cacao. AB - Phvytophthora megakarya is a devastating oomycete pathogen that causes black pod disease in cacao. Phytophthora species produce a protein that has a similar sequence to the necrosis and ethylene inducing protein (Nep1) of Fusarium oxysporum. Multiple copies of NEP1 orthologs (PmegNEP) have been identified in P. megakarya and four other Phytophthora species (P. citrophthora, P. capsici, P. palmivora, and P. sojae). Genome database searches confirmed the existence of multiple copies of NEP1 orthologs in P. sojae and P. ramorum. In this study, nine different PmegNEP orthologs from P. megakarya strain Mk-1 were identified and analyzed. Of these nine orthologs, six were expressed in mycelium and in P. megakarya zoospore-infected cacao leaf tissue. The remaining two clones are either regulated differently, or are nonfunctional genes. Sequence analysis revealed that six PmegNEP orthologs were organized in two clusters of three orthologs each in the P. megakarya genome. Evidence is presented for the instability in the P. megakarya genome resulting from duplications, inversions, and fused genes resulting in multiple NEP1 orthologs. Traits characteristic of the Phytophthora genome, such as the clustering of NEP1 orthologs, the lack of CATT and TATA boxes, the lack of introns, and the short distance between ORFs were also observed. PMID- 16353638 TI - Damage to DNA and lipids in Boletus edulis exposed to heavy metals. AB - This study investigates the potential of emissions from a zinc smelter to induce oxidative damage to DNA and lipids in Boletus edulis, the king bolete. Concentrations of cadmium, zinc, copper, and mercury were determined in 16 fruit bodies collected near the smelter (exposed group), as well as in 15 reference samples. Frequency of apurinic/apyrimidinic (AP) sites in DNA (a pre-mutagenic DNA base modification) and concentration of lipid hydroperoxides were chosen as damage parameters. Concentrations of the four metals, as well as oxidative damage to DNA and lipids were significantly elevated in the exposed group (Mann-Whitney, P<0.001). Both damage parameters correlated positively with concentrations of cadmium, zinc or copper in fruiting bodies (Spearman's P<0.01). Frequency of AP sites correlated significantly with mercury in the fruit bodies (P<0.05), whereas the association between lipid hydroperoxides and mercury was insignificant. Frequency of AP sites correlated positively with concentration of lipid hydroperoxides (P<0.001). Negative trends for the associations between concentrations of metals and AP sites or lipid hydroperoxides in the reference group (significant only for mercury and lipid hydroperoxides; P<0.05) suggest that in B. edulis low concentrations of mercury, possibly also of other of the metals determined in the present study, may induce dose-response relationships of a hormetic ('J-shaped') nature. PMID- 16353639 TI - Screening of antimicrobial activities in Trichoderma isolates representing three trichoderma sections. AB - Methanol extracts from 24 Trichoderma isolates, selected as biocontrol agents and representating different species and genotypes from three of the four taxonomic sections of this genus (T. sect. Trichoderma, T. sect. Pachybasium and T. sect. Longibrachiatum) were screened for antibacterial, anti-yeast and antifungal activities against a panel of seven bacteria, seven yeasts and six filamentous fungi previously used in similar studies. Two different growth media were tested (potato dextrose broth and CYS80), and all isolates included in the antimicrobial tests showed at least one inhibitory activity against one of the target microorganisms in one of the two culture media. No statistically significant differences were detected in the number of active strains between the two culture media, but the highest number of inhibitory strains against bacteria and fungi were found in strains from Trichoderma sect. Pachybasium, whereas strains from T. sect. Longibrachiatum showed the highest anti-yeast values. In all cases, a correlation was found between the strains that were active against yeasts and fungi. However, some degree of variability was detected for strains within the same taxonomic section. In general terms, strains from T. asperellum (mainly in CYS80 medium), and T. longibrachiatum gave the best non-enzymatic antimicrobial profiles. PMID- 16353640 TI - Molecular evidence confirms multiple origins of aquatic hyphomycetes. AB - Traditional taxonomy of aquatic hyphomycetes has been based on conidial morphology and development. Since the predominantly tetraradiate and sigmoid forms are due to convergent evolution, they are often phylogenetically non informative. The comparison of nuclear small-subunit ribosomal DNA sequences of 30 species (22 new, eight previously published) assigned 22 to Leotiomycetes, four to Dothideomycetes, three to Sordariomycetes, and one to Orbiliomycetes. Eight species of Anguillospora were distributed among the Leotiomycetes, Dothideomycetes, and Orbiliomycetes. All three anamorphs connected with Massarina were assigned to the Pleosporales, however, Clavariopsis aquatica and Tumularia aquatica separated from Anguillospora longissima. The nSSU rDNA sequences of several species were identical (e.g. Anguillospora crassa and A. furtiva), suggesting the need to include less conservative genes for resolving such differences. PMID- 16353641 TI - Fungal associates of Salix repens in northern oceanic Britain and their conservation significance. AB - Macromycetes recorded from beds of creeping willow, Salix repens, in the northern archipelagos of Shetland and Orkney are listed and discussed. Two components are demonstrated, an arctic-alpine group and one which is more typical of lowland frondose woodland communities in mainland Britain. The importance of maintaining, even encouraging the development of creeping willow beds with their attendant ectomycorrhizal fungi in conservation terms are explored. PMID- 16353642 TI - The occurrence and distribution of Piptocephalis, syncephalis and Chaetocladium species on dung. AB - Records of the Piptocephalidaceae and Chaetocladiaceae from over 800 samples of dung collected worldwide are analysed. Piptocephalis species were frequent, occurring especially on dung of rabbit (31% of all samples) in temperate regions. Chaetocladium and Syncephalis species were much less frequent. The status of these fungi as coprophils and reasons for the differences in distribution and substrate preference are discussed. PMID- 16353643 TI - Emergencies in the office. PMID- 16353644 TI - Managing emergencies Part 1. PMID- 16353645 TI - A 2-year-old girl with an abnormal gait. PMID- 16353646 TI - A 9-year-old boy with facial lesions. PMID- 16353647 TI - Evaluating cardiovascular presentations: what does an electrocardiogram have to offer? AB - The general pediatrician remains the key to the timely recognition and treatment of cardiovascular disorders, particularly those that present acutely and may require immediate attention. In the evaluation of these cardiovascular urgencies, ancillary studies such as the electrocardiogram continue to be important, readily available tools that can aide in the diagnostic process. It is thus incumbent on the general practitioner to foster the skill necessary to employ such tools reliably, particularly in the setting of urgent evaluations. At the same time, recognition of the limitations of such testing will help both in the acute setting and in the understanding of their application in population settings. PMID- 16353648 TI - Endocrine emergencies: recognizing signs and symptoms. AB - Endocrine emergencies may present to the pediatric practitioner in the office setting in a variety of forms. Four of the more common pediatric endocrine emergencies (DKA, symptomatic hypoglycemia, adrenal insufficiency, and hypocalcemia) have been discussed here. The recommended approach to a child or adolescent with an endocrine emergency involves recognizing clinical disease, stabilizing the patient with basic and advanced life support intervention, and transferring the patient to a facility which can provide appropriate definitive care. PMID- 16353649 TI - Rash, fever, and neck pain in the office: is this a neurologic emergency? AB - Anticonvulsants, neuroleptics, and antispasticity agents are used with increasing frequency in the pediatric population. Each of the drugs discussed in this article has serious but potentially reversible adverse effects. Pediatric primary care providers must be aware of the potential emergencies associated with the use of these neurologic medications to provide prompt and effective treatment. PMID- 16353650 TI - Respiratory distress. AB - Preparation for pediatric pulmonary emergencies in the office setting includes adequate training for all medical staff, properly sized and working equipment, and medications to help alleviate respiratory distress when indicated. Status asthmaticus, viral bronchiolitis, and croup account for the vast majority of respiratory emergencies encountered in the pediatric office setting. Timely application of proven approaches to assessment and treatment of these illnesses can prevent hospitalization, decrease length of hospitalizations, and save lives. PMID- 16353651 TI - Emergencies in adolescents: management guidelines for four presentations. AB - The cases presented in this article are representative of adolescent emergencies that may be encountered by the office practitioner. Many more examples in the area of adolescent gynecology could have been presented, including ectopic pregnancy and pelvic inflammatory disease. Substance abuse issues will be addressed in the second part of this topic, which will be published in the December 2005 issue of Pediatric Annals. Consultation with an adolescent medicine specialist is always recommended for challenging cases. PMID- 16353652 TI - A surgeon in your corner. AB - The surgical emergencies presented in this article encompass the vast majority of the nontraumatic emergencies seen by a pediatric surgeon but represent only a small proportion of surgical disease in children. Most children seeking outpatient medical care do not need a surgeon, but those who do need one quickly. When one of the processes detailed above is suspected, it is always best to avail oneself of the opinion of a surgeon in a timely manner. PMID- 16353653 TI - One more river to cross--looking back, moving forward: advancing the NMA "equality agenda" in the era of 21st-century medicine. AB - The following is an excerpt of the inaugural address given July 27, 2005 by Sandra L. Gadson, MD, who was installed as the 106th president of the National Medical Association (NMA) during NMA's 2005 Annual Convention and Scientific Assembly in New York, NY. PMID- 16353654 TI - NMA calls response to Hurricane Katrina a "national disgrace". AB - The National Medical Association is the oldest and largest organization representing the interest of more than 30,000 African-American physicians. The NMA was founded in 1895. PMID- 16353655 TI - Inpatient surgical treatment patterns for patients with uterine fibroids in the United States, 1998-2002. AB - OBJECTIVE: To analyze the impact of patient and organizational characteristics on surgical treatment patterns for patients with uterine fibroids. METHODS: Unadjusted means and percentages were calculated from a population-based inpatient sample (HCUPNIS). Multiple logistic regression analysis was used to estimate the prevalence odds ratios for the association of uterine fibroid treatments and covariates of interest. RESULTS: More than 1.2 million patients with a primary diagnosis of uterine fibroids were treated from 1998 to 2002. Of these, 84.4% received a hysterectomy and 12.3% received a myomectomy. Total abdominal hysterectomy was the most common procedure. The number of supracervical hysterectomies increased 18.1% over the five-year period. Black women and Asians/Pacific Islanders were more likely than white women to receive a myomectomy. All types of hysterectomies were more common in Medicaid patients compared with private/HMO patients. With the exception of patients in ZIP codes with a median income of <$25,000 per year, an inverse relationship was identified between income and hysterectomy rates. CONCLUSIONS: The management of uterine fibroids appears to differ across a variety of socioeconomic factors and institutional characteristics. This study suggests that additional research should be conducted to assess the impact of nonclinical factors on treatment decisions for patients with uterine fibroids. PMID- 16353656 TI - Fertility outcomes following myomectomy in an urban hospital setting. AB - OBJECTIVE: Infertility is rarely a consequence of myomas. However, a causal relationship may be suspected when other causes of infertility have been excluded. Uterine myomas have been reported in 27% of infertile women; 50% of women with unexplained infertility become pregnant after myomectomy. The objective of this study was to establish the impact of the surgical removal of myomas on fertility outcomes in women experiencing recurrent pregnancy loss or unexplained infertility. Fallopian tube, anovulatory disorders and male fertility factors had been appropriately excluded. DESIGN: This was a retrospective study in which we compiled data from the medical records of eight patients from 2003 2004 who underwent abdominal myomectomy for infertility or recurrent pregnancy loss. We calculated rates for subsequent spontaneous abortion, preterm delivery, cesarean delivery, malpresentation and postpartum hemorrhage. RESULTS: There were two patients who were nulliparous premyomectomy, and six had recurrent pregnancy losses. There was a cumulative success rate of 75% (six live births in eight patients) following myomectomy. One had two subsequent pregnancies. There were no spontaneous abortions. Three (37.5%) patients failed to conceive postmyomectomy, one of which was found to have bilateral tubal occlusion. Of the six pregnancies achieved, two (33%, 95% CI 2.06, 3.14) were preterm deliveries, six (100%, 95% CI 1.74, 3.50) were delivered by cesarean section and three (50%, 95% CI 3.50, 1.73) were malpresentations (two breech, one transverse lie). One patient (16%, 95% CI 2.06, 3.30) had abruptio placentae and two patients (33%, 95% CI 2.06, 3.14) experienced postpartum hemorrhage. CONCLUSION: This study suggests that there may be a beneficial effect of surgical removal of myomas on enhancing fertility and successful pregnancy outcome. However, the sample was too small to achieve statistical significance. PMID- 16353657 TI - Evaluating a tailored intervention to increase screening mammography in an urban area. AB - METHOD: The study was conducted over a four-year period, 1996 and 2000. Participants were recruited using Computer Assisted Telephone Interviewing (CATI) software and random-digit dialing (RDD). Study eligibility criteria included living in the King/Drew Medical Center service area in Los Angeles, having an operable telephone, being female > or = 40 years old and not having had a screening mammogram in the past year. Four-hundred-thirty respondents were randomly assigned to the intervention and comparison groups. English and Spanish focus-group-tested tailored interventions were administered telephonically by trained interviewers. African Americans and Latinas constituted 83.0% of the sample at assignment and 83.8% at six-month follow-up, which is representative of the study area. RESULTS: The main outcome variable of interest in this study was having a screening mammogram during the time interval between baseline and the six-month follow-up assessment. Multiple logistic regressions that revealed factors predicting the outcome variable included: 1) age (p < or = 0.05, OR=2.22, CI 0.98-5.0); 2) study group (p < or = 0.05, OR=1.76, CI 1.06-2.92); 3) prior mammograms (p < or = 0.05, O0R=2.51, 1.39-4.56); and 4) and knowledge of the age when a woman should begin getting mammograms on a regular basis (p < or = 0.05, OR=0.55, 0.33-0.92). CONCLUSION: Tailored telephone counseling increased the instances of screening mammograms by nearly 8% in the intervention group at follow-up. The results of this study confirm previous findings regarding the impact of structural and behavioral factors related to screening mammography. PMID- 16353658 TI - Racial differences in the physical and psychosocial health among black and white women with chronic pain. AB - Gender-based differences in pain epidemiology, pain threshold, attitudes toward pain management, coping styles and social roles are well described, yet little is known about the chronic pain experience in women or the role race plays. A retrospective analysis of self-reported data using a secondary clinical database was performed to elucidate the relationship between race and pain severity, depression, physical disability, posttraumatic stress disorder (PTSD) as well as affective distress in women with chronic pain. White (n=1,088) and black (n=104) adult women were compared based on their responses to the McGill Pain Questionnaire, Beck Depression Inventory, Pain Disability Index, Posttraumatic Chronic Pain Test and items from the West-Haven Yale Multidisciplinary Pain Inventory. After accounting for sociodemographic, medical, psychological and physical confounders, there was no significant race effect for pain severity or affective distress. However, black women with chronic pain experience more physical impairments than white women with chronic pain (beta = 4.622; p<0.005). Except for the family/home responsibilities, similar differences were found on all PDI subscales. We also found that disability mediates the race-depression relationship such that black women are comparatively more vulnerable to depression as a result of higher disability. Due to the economic, social and emotional impact that disability has on women with chronic pain and their families, these findings have significant implications for chronic pain research as well as its management in black women. PMID- 16353659 TI - Knowledge, attitudes and beliefs of women about the importance of prostate cancer screening. AB - OBJECTIVES: To understand women's knowledge, attitudes and beliefs about prostate cancer. METHODS: A survey was self-administered to 324 women age >18 years. It contained 42 questions that assessed women's knowledge about prostate cancer, possible risk factors, and opinions regarding screening and early detection. Women were grouped as married or unmarried for convenient comparisons. Chi squared and F statistics were performed. RESULTS: Ninety-seven percent of married women reported having some influence over the healthcare decisions of their spouse. Married women's worst fear about their spouse or family member's diagnosis of prostate cancer was death. The most important benefit of prostate cancer screening was the possibility of cure, while the main hindrance was fear of the digital rectal exam. Marital status, age, educational level and income were all significantly associated with women's knowledge about prostate cancer (p<0.001). CONCLUSIONS: Women play an important role in health-related matters in the home. Educating women on prostate cancer may improve early detection efforts and reduce the devastating impact of this disease on their family. PMID- 16353660 TI - Characterization of frequent douchers attending a community clinic primarily serving African-American women. AB - OBJECTIVE: The purpose of this study is to characterize African-American women attending a community clinic who report frequent douching (douching > or = 2 times per week). METHODS: A consecutive sample of 115 black women attending a community clinic were interviewed face-to-face about their douching practices. Logistic regression was used to control for age and compute odds ratios and 95% confidence intervals. RESULTS: Of the 115 women interviewed, 93% (107) had douched sometime during their lifetime; 16% (18) reported douching > or = 2 times per week. Frequent douchers compared with women who douche < 2 times per month were more likely to report douching after sex [89% (n=16) vs. 49% (n=32), odds ratio (OR): 5.35, 95% confidence interval (CI): 1.09, 26.2] or after discharge [89% (n=1 6) vs. 58% (n=38), OR: 8.11, 95% CI: 1.64, 40.1], and self-report a history of gonorrhea [28% (n=5) vs. 8% (n=5), OR: 4.87, 95% CI: 1.07, 22.2]. CONCLUSION: Further research should be done to understand the use of douching as an STD/HIV prevention method and the association between sexual risk behaviors and douching practices. PMID- 16353661 TI - Reproductive decision-making among HIV-Infected women. AB - OBJECTIVE: To describe factors related to reproductive decision-making among HIV infected women. MATERIALS AND METHODS: A sample of HIV-infected women (N=104) who received care at an HIV clinic in the southern United States were interviewed about their reproductive decision-making. Women who became pregnant subsequent to HIV diagnosis were compared to women who did not become pregnant, and women who underwent a sterilization procedure subsequent to HIV diagnosis were compared to women who did not get sterilized. RESULTS: Compared to women who did not get pregnant after receiving an HIV diagnosis, women who became pregnant were more likely to be young, single, diagnosed earlier in the epidemic and to have more recently used a noninjecting drug. Among women who did not get pregnant, 63% reported their diagnosis greatly affected that decision. Having a partner who wants more children was not associated with pregnancy. Compared to women who did not get sterilized after learning their HIV status, women who did get sterilized tended to be Baptist and already had a prior live birth. Neither a woman's desire nor her partner's desire for more children was associated with sterilization. CONCLUSIONS: HIV is an important influence on HIV-infected women's reproductive choices, regardless of the decision being made. Reproductive counseling by HIV care providers needs to be sensitive to all the issues faced by these women. PMID- 16353662 TI - Pregnancy complicated by emphysematous pyonephrosis. AB - Emphysematous pyonephrosis is a rare, rapidly progressive infection characterized by the presence of exudative material and fermentative gases within a dilated renal collecting system. This unusual clinical entity is noted almost exclusively in diabetic patients, often in conjunction with chronic urinary tract infection and obstruction. In most instances, E. coli is the etiologic agent; however other enterobacteriaceae may be isolated. High fever, leukocytosis, renal parenchymal necrosis and the accumulation of gases in the kidney and perinephric tissues are the hallmark of this condition. Surgical resection of the involved tissue and systemic antibiotic administration are among the primary mainstays of therapy. We present a unique case of a pregnant patient with fulminant emphysematous pyonephrosis and the successful treatment rendered. PMID- 16353663 TI - Long-term pegylated liposomal doxorubicin use in recurrent ovarian carcinoma. AB - BACKGROUND: Ovarian carcinoma is a devastating disease because patients are diagnosed with advanced disease at presentation and five-year survival ranges from 5-20%. Salvage therapy becomes important for survival in those patients with recurrent disease. There are a variety of agents with relatively similar response rates; however, side effects may limit choice. Pegylated liposomal doxorubicin was found to be less toxic but as effective as other agents. Cardiotoxicity continues to be a concern with long-term antracycline use. CASES: We present three cases of women diagnosed with advanced ovarian carcinoma. Each patient initially underwent optimal cytoreductive surgery, however, developed recurrent disease and were treated with pegylated liposomal doxorubicin. One patient remains disease-free following complete response. Two patients were maintained on pegylated liposomal doxorubicin with stable disease for 18 and 34 months, respectively. These cases demonstrate that pegylated liposomal doxorubicin can be used for extended periods of time without cardiotoxicity. CONCLUSION: The adverse events were few with cumulative doses as high as 1,360 mg/m2. These cases show that pegylated liposomal doxorubicin may be a promising agent in recurrent ovarian carcinoma. We recognize the limitations of our data. The results need to be confirmed in a larger group of patients. PMID- 16353664 TI - Reducing HIV/AIDS transmission among African-American females: is the female condom a solution? AB - Rates of HIV/AIDS have increased at an alarming rate among minority women, especially African-American women. Suggestions that have been presented to decrease HIV/AIDS transmission among African-American women include promoting abstinence and the use of the male condom. Little recognition and support have been given for promoting the female condom as a viable solution to combating the HIV/AIDS epidemic. PMID- 16353665 TI - Sickle cell anemia and academic achievement in Africa. PMID- 16353666 TI - Legitimacy for traditional healers? PMID- 16353667 TI - Critical tests missing from July case report and diagnosis of primary lymphedema tarda. PMID- 16353668 TI - Steroidogenic factor 1 gene transcription is inhibited by transforming growth factor beta. AB - The orphan nuclear receptor, steroidogenic factor 1 (SF-1), plays a major role in adrenal and gonadal development, as well as in sexual differentiation. It has been demonstrated that the expression of a number of genes regulated by SF-1 is inhibited by the transforming growth factor, (TGF-beta). To date, however, the influence of TGF-beta on the expression of SF-1 gene has not been reported. A Northern blot analysis with the use of a radiolabeled cDNA probe, and immunodetection with antibodies directed against SF-1, demonstrated that the Sf-1 transcript and the SF-1 protein levels were lowered by TGF-beta in Y-1 adrenocortical cells, both in untreated and adenylyl cyclase activator, forskolin treated cells. An examination of the Sf-1 transcript stability in the presence of actinomycin D revealed no influence of TGF-beta on the rate of Sf-1 mRNA decay. Inhibition of Sf-1 expression by TGF-beta was abolished by cycloheximide, suggesting that the growth factor inhibitory effect requires ongoing protein synthesis. We conclude that in Y-1 cells TGF-beta inhibits the expression of SF-1 gene at a transcriptional level, and we postulate that the inhibitory effect of TGF-beta on steroid hormone synthesis in the adrenal cortex could be due to an attenuated transcription of Sf-1. PMID- 16353669 TI - Effects of streptozotocin-diabetes and insulin replacement on the epididymis of prepubertal rats: histological and histomorphometric studies. AB - The present study evaluates the effects of streptozotocin (STZ)-induced diabetes and insulin replacement on the histoarchitecture of caput, corpus, and caudal epididymides during the critical period of sexual maturation in rats. Prepubertal male Wistar rats (40 days old) were made diabetic by a single injection of STZ (120 mg/kg body weight, intraperitoneally). To one set of diabetic rats, insulin was replaced daily at a dose of 3 U/100g body weight, subcutaneously in two equally divided doses at 8:00 a.m. and 6:00 p.m. All the rats were killed on the 61st day of postnatal life. STZ-diabetes reduced the body weight and also caused regression of epididymis, leading to a decrease in the absolute weight of caput, corpus, and caudal regions. Histological studies also revealed a considerable reduction in the size of the tubule and lumen of these segments with an increase in interstitial stroma. Because of shrinkage of tubules, principal cells were packed tightly with clumping of nuclei. Stereological studies support atrophic changes in the caput, corpus, and caudal epididymides by reduction in tubular diameter, volume, and surface density. The epididymal lumen of STZ-treated rats was totally devoid of spermatozoa. These findings emphasize the detrimental effects of diabetes on the maintenance and establishment of fully differentiated epididymal epithelium during sexual maturation. Insulin replacement was only able to prevent the adverse effects of diabetes on certain parameters and this response was region-specific. PMID- 16353670 TI - Growth restriction and exendin 4 promote endocrine expression in cultured islet cells derived from patients with persistent hyperinsulinemic hypoglycemia of infancy (PHHI). AB - Islets derived from patients with persistent hyperinsulinemic hypoglycemia of infancy, PHHI, show a significant capacity to proliferate in vitro without the addition of growth factors. However, as with other differentiated cells, PHHI derived islet cells show a loss of differentiated function with repeated subculture. Here, we have investigated methods of extending the differentiated function of PHHI-derived endocrine cells following in vitro expansion. The experiments were carried out on 13 primary pancreatic cell cultures from patients with PHHI, the majority of which (n = 11) were glucose unresponsive--a distinctive feature of PHHI disease. After a 20-day period of cell expansion in 10% FCS, cells were switched to media containing varying concentrations of FCS with or without exendin 4 and endocrine function was measured using ELISA and RT PCR for insulin and PDX-1. Switching the expanded cultures to low serum was shown to slow cell division while maintaining the residual differentiated endocrine characteristics of all the cultures tested. Exendin 4 was shown to further enhance the improved insulin secretion shown by low serum cultures, although in glucose nonresponsive cells, this was at the expense of insulin stores. However, we did observe that exendin 4 could upregulate insulin secretion, insulin storage, and PDX-1 expression in glucose responsive PHHI cultures. PMID- 16353671 TI - Plasma leptin concentrations in postmenopausal women with osteoporosis. AB - Osteoporosis is less common in obese individuals with increased bone mineral density (BMD) and plasma leptin concentrations. The aim of this study was to determine the correlation between leptin levels and BMD in postmenopausal women. The study consisted of 90 postmenopausal women with a mean age of 53.45 +/- 0.87 years who visited our outpatient clinic for the evaluation of BMD. Thirty-six post-menopausal women with osteoporosis (mean age: 54.52 +/- 1.41 years and mean body mass index (BMI, kg/m2) 29.33 +/- 0.66), 30 age- and BMI-matched postmenopausal women with normal BMD, and 24 postmenopausal women (mean age: 52.79 +/- 1.48 years and mean BMI: 29.45 +/- 0.89) with osteopenic BMD were included in the study. Plasma concentrations of leptin after an overnight fast were measured by radioimmunoassay. BMD values were measured by dual-energy X-ray absorptiometry (DEXA) at the L2-L4 lumbar spine and femoral neck. The median spine BMD value in the patient group (0.67 +/- 0.08 g/cm2, mean +/- SEM) was significantly lower than that in the control group (1.02 +/- 0.25 g/cm2, mean +/- SEM) and osteopenic group (0.87 +/- 0.05 g/cm2, mean +/- SEM) (p < 0.005). The mean spine BMD value (T score -3.63 +/- 0.25, mean +/- SEM) and the mean femur neck BMD value (T score -2.55 +/- 0.18, mean +/- SEM) of the osteoporosis group were significantly lower than that in the normal BMD group (+ 0.33 +/- 0.14 and + 0.27 +/- 0.18, P < 0.001) and in the osteopenia group (-1.74 +/- 0,1 and -1.18 +/ 0.05, p < 0.005). The mean plasma leptin concentration in the osteoporotic group (17.03 +/- 1.40 ng/ml) was not significantly different from that in the normal BMD group and the osteopenia group (16.55 +/- 1.50 ng/ml; 16.16 +/- 1.60, respectively, p > 0.150). Plasma leptin concentrations were correlated with BMI in three groups (r(s) = 0.450, p = 0.025 in normal BMD group and r(s) = 0.4254, P = 0.009 in the osteoporotic group, and r(s) = 395, p = 0.015 in the osteopenia group. There was no correlation between plasma leptin concentrations and BMD values in three groups (r(s) = -0.89 in normal BMD group, r(s) = -0.124 in osteopenia group, and r(s) = -0.195 in osteoporosis group). From this study we conclude that circulating plasma leptin does not have a significant direct influence on bone mass in postmenopausal women. PMID- 16353672 TI - The effect of testosterone supplement on insulin sensitivity, glucose effectiveness, and acute insulin response after glucose load in male type 2 diabetics. AB - Our understanding of the effect of androgens on insulin action and glucose metabolism is incomplete. Several different models and methods have been used to study androgen effects, with some studies indicating that higher testosterone levels are associated with increased insulin resistance. In polycystic ovary syndrome, where high testosterone levels are frequently found, affected patients have a higher risk of diabetes. In contrast, increased insulin resistance was found in both hypergonadotropic and hypogonadotropic men with hypoandrogenism, patients with Klinefelter's syndrome and men with idiopathic gonadotropin deficiency. Insulin resistance is considered to be one of the cornerstones in the state that ultimately leads to clinically established type 2 diabetes mellitus. In addition, men with type 2 diabetes have relative hypogonadism. Therefore, supplementation with testosterone might play a role in improving both insulin resistance and hypogonadism. The study population consisted of 11 male patients with type 2 diabetes. Their mean age was 57.7 +/- 3.41 years, the body mass index (BMI) was 24.4 +/- 1.02 kg/m2, and the waist-to-hip ratio (W/H) was 0.91 +/- 0.05. The patients were all treated with oral hypoglycemic agents. The men received androgen injections every 3 weeks intramuscularly for 12 weeks. The injections were testosterone depot 100 mg/3 weeks. Insulin sensitivity, glucose effectiveness and area under acute insulin response were calculated from "minimal model" algorithms. There were no significant differences in the value of BMI, W/H ratios, plasma lipid concentrations, testosterone, homeostasis model assessment (HOMA) of insulin sensitivity, and beta-cell function, before and after supplementation of testosterone. Furthermore, the insulin sensitivity (SI) (1.04 +/- 0.25, 1.11 +/- 0.36 x 10(-5) min(-1/)pM; p = 0.43), glucose effectiveness (EG) (0.018 +/- 0.003, 0.017 +/- 0.002 min(-1); p = 0.29), and acute insulin response (AIR) after a glucose load (45.7 +/- 24.3, 50.1 +/- 32.5 pM; p = 0.45) did not change significantly after supplmentation with testosterone. In our study, there was no improvement of SI, EG, and AIR after 3 months of Testosterone Depot treatment in type 2 diabetes, but we believe that duration and dosage of the androgen therapy might play an important role in improving insulin sensitivity. The mechanisms by which testosterone causes insulin resistance is unknown, and larger studies on androgen treatment in type 2 diabetic patients are necessary. PMID- 16353673 TI - The capability of glucose toxicity on severe type 2 diabetes. AB - The underlying defects of type 2 diabetes are impaired insulin sensitivity and decreased beta-cell function. In poorly controlled type 2 diabetes, the 'glucose toxicity' further deteriorates these defects. The objective was to determine whether correction of glucose toxicity will lead to improve insulin sensitivity and beta-cell function in these severely affected patients. Twelve severe type 2 diabetic patients were enrolled. An intravenous glucose tolerance test was performed before and after treatment with insulin for 3 months. The insulin sensitivity, glucose sensitivity, and acute insulin response after glucose loading were calculated by a minimal model algorithm. The lipid profiles did not change significantly after insulin therapy, but the hemoglobin A1c, level improved significantly (12.2 +/- 2.2% to 9.2 +/- 1.9%; p = 0.001). The insulin sensitivity, glucose sensitivity, and acute insulin response did not change significantly with insulin therapy. Correction of hyperglycemia with 3-month insulin therapy may improve metabolic effect instead of insulin sensitivity, glucose sensitivity, and acute insulin response to glucose load in severe type 2 diabetic patients. PMID- 16353674 TI - [Occurrence of Giardia and Cryptosporidium in wastewater, surface water and ground water samples in Palermo (Sicily)]. AB - This paper reports the investigation carried out on the occurrence of Cryptosporidium and Giardia (oo)cysts in water samples of two municipal treatment plants, and in surface water and ground water wells of the province of Palermo. The wastewater samples taken before and after treatment process were assayed over the course of one year Giardia cysts were detected in all samples throught the year at higher concentration levels than Cryptosporidium oocysts, with a peak observed in spring. The overall removal efficiency of (oo)cysts in the treatment plants was about of 90%. Their presence were also searched in surface waters (three artificial lakes and one river); (oo)cysts were detected in one lake at very low concentration; on the contrary, both parasites were found at high concentration levels in all the samples collected throught one year from the water of the river. The pattern of occurrence of both parasites appears temporally related to the level of rainfall trend. Cryptosporidium and Giardia were also found in ground water wells; their presence occurred only in waters taken from wells at a depth lower than 31 meters with concomitant presence of faecal bacteria. These results may provide further insight into the possible source of Cryptosporidium and Giardia in natural environmental and stress the potential risk associated with the use of waters for recreational and agricultural purposes. PMID- 16353675 TI - [Monitoring and control of opportunistic bacteria in a spa water used for aerosol hydrotherapy]. AB - A bicarbonate-sulphate-calcic water of a therapeutic spa was monitored for the presence of Legionella, Pseudomonas and Mycobacteria. The water was analysed by taking samples from the well, the feed tank and from the final aerosol generating devices of two different water lines, the former at 21-23 degrees, the second at 36-38 degrees. The bacteria in question were always absent from the well. Legionellae were found in the water of aerosol equipment: Legionella micdadei was isolated from 75% of samples, L. bozemanii from 75% and 50% (respectively 36-38 degrees and 21-22 degrees water lines) and other species of environmental Legionellae from 25% of samples. The water of aerosol equipment presented high total bacterial counts (10(3)-10(4) cfu/ml) and exspecially high concentrations of Pseudomonadaceae (10(2)-10(3) cfu/100 ml). These bacteria, unlike the Legionellae, were also isolated from the feed tank at mean concentrations of about 10(2) cfu/100 ml. Mycobacteria were found in 75 and 50% of samples collected from final devices, respectively from the heated and not heated water lines. The isolates were M. gordonae (85% of isolates) and M. fortuitum (15%), but at concentrations very low. Both treatments with sodium hypochlorite (20 ppm of residual chlorine) and peracetic acid (20 ppm) resulted in the reduction of total bacterial counts and elimination of Pseudomonas from the water in the tank, but not in elimination of Pseudomonas and Legionellae from the nebulizers. The disinfectants were evidently not able to efficiently reach all the points where Pseudomonas and Legionella had settled and grown. In order to obtain total abatement it was necessary to carry out a radical restructuring of the plant, involving the replacement of the old nebulizer benches with new aerosol equipment that could be subjected to a new system of programmed thermal shock. PMID- 16353676 TI - [Biological risk in the operating room: microbiological monitoring of the environment and analysis of the associated variables]. AB - The authors present an environmental microbiological monitoring programme carried out over a period of 15 months in 16 operating theatres performing specific types of surgery. The levels of microbial contamination of the air and of four of the most representative surfaces of the clean area were determined at 3 different times for each theatre, both before and during surgery. For the air assessment, the results obtained with three different samplers, Sed-3 Unit, SAS and RCS, were compared. The results were on the whole acceptable, but some poor conditions were detected during the theatres in use, especially in general surgery theatres; in some of these the floors showed levels of contamination consistently exceeding the reference limits. As the monitoring programme proceeded, the microbiological quality of the air and of the surfaces in the theatres notably improved. The three air samplers showed different conditions expressed with units of measure not always readily comparable. For active samplers, the bacterial load determined by RCS, although less variable, were always higher (even 2-3 fold) than those obtained with the SAS. Passive sampling takes longer but determines the real risk of infection for the patients; contemporary determination of the fall-out and the CFU/m3 helps to identify the occupational risks. Since the limit values established by the ISPESL guidelines for the operating theatres have been defined only for active samplers, there is urgent need for more exhaustive national guidelines to define similar values also for passive sampling. The Authors conclude stressing the importance of promoting continuing information-education programmes to heighten the awareness of all those involved in operating theatre activities. PMID- 16353677 TI - [Preoperative hair removal review]. AB - Surgical site infection (SSI) frequency is influenced by several risk factors, among which perioperative hair removal. Traditionally it is part of the procedures carried out on skin before surgery. Although preoperative shaving is widely used in many hospitals, its use should be re-evaluated. Therefore the AA. carried out a systematic literature review about the opportunity, the modality and the risks associated to preoperative hair removal. We found first of all that preoperative shaving (razor) is associated with a significantly higher surgical site infection (SSI) risk. Second, when hair removal is considered necessary by the surgeon, it should be carried out by means of a clipper or depilatory cream. Furthermore hair removal should be performed immediately before surgery and not in advance, to avoid an increased risk of SSI. PMID- 16353678 TI - [Epidemiology of hospital loads in patients with multiple hospitalizations due to diabetes]. AB - This study describes the results of a retrospective study carried out to evaluate the hospitalizations of patients with type 2 diabetes admitted from January to June 2001 in the teaching hospital "SS. Annunziata" of Chieti. This research evaluates the generic appropriateness using the PRUO manual and the specific appropriateness using the guidelines approved by Italian Diabetes Association, Italian Diabetes Society and Italian College of General Practitioners. A sample of 196 medical charts was examined. The percentage of inappropriate admission was 21.9%. The "critical" clinical conditions of patients were responsible for only 23.7% of inappropriate admissions. The first reason of the inappropriateness of the admission was the execution of diagnostic examinations (60.5%), followed by the execution of medical therapy (23.2%) and waiting for surgical intervention (16.3%). 46.5% of inappropriate hospitalization was prescribed by specialists. Concerning specific appropriateness, 42.3% of hospitalization was inappropriate. These findings suggest that a system for the assessment of disease management of diabetes should be started up in the Abruzzo region. Moreover, guidelines utilization should be implemented in order to get a more correct utilization of acute hospital by specialists and GPs. PMID- 16353679 TI - [Prevalence of obesity, overweight and hypertension in children and adolescents from Abruzzo, Italy]. AB - The present survey was aimed at determining the prevalence of overweight, obesity, hypertension, their correlation and the association with gender, age and provenience, in the pediatric population of Pescara province, Italy. During the academic year 2001-2002, the body mass index (BMI) and blood pressure of 15.612 scholars aged 6-14 years (mean age 9.9), coming from all province schools, have been measured. Two scales have been used to define obesity and overweight status (one developed by the National Center for Health Statistics--NCHS--the other by Cacciari et al.); while hypertension has been attributed using National Institute of Health curves. Several multiple logistic regression models have been fitted to analyze data. According to NCHS standards, the prevalence of childhood and adolescence overweight and obesity was, respectively, 40.6% and 19.5%. Using curves by Cacciari, the prevalence were 33.3% and 7.7%. Hypertensive subjects were 11.1%. With both scales, the prevalence of weight problems widely decreased after 12 years of age, and either obesity or overweight were significantly related with male sex, hypertension and rural area residence. A strong independent association was found between living inside the city and hypertension. Although the differences in the dimension of the problem "overweight" according to the used scale, and a potential overestimation of hypertension must be taken into account, the overall data suggest an urgent need, especially for primary school children, of preventive interventions aimed at reducing both overweight and blood pressure. PMID- 16353680 TI - [Customer satisfaction analysis in women attending an organized mammographic screening. Pilot study at Trento]. AB - Present paper reports on the results of a pilot customer satisfaction study carried out on 1.720 consecutive women (18.7 with spontaneous access) attended senology department (Trento and Borgo) in the context of a organized mammography screening programme, started in October 2000. Data were collected by a questionnarie filled by the women after mammography. 4.3% of the invited women reported the receiving the letter late, 0.9% considered it inaccurate, 7.2% had some problems for getting a new date for test, 1.8% of the whole sample reported some problems for external access and 2.2% for internal access to health facilities; 1.9% perceived waiting room as not friendly. The welcoming by the personnel is judged quite well, only 0.6% complained about it. Trento centre, is more efficient than Borgo. In 21.7% of the cases the mammography has been performed within next 30 minutes of the fixed time and in 7.9% besides that. 36.8% of the whole sample perceived mammography as tiresome and 4.1% as painful. The percentage of women reporting mammography disconforting increase, in Trento sample, according the education level as previously reported. The data about perceived quality are satisfactory, on the whole. Anyway it would be opportune to contain the waiting time. PMID- 16353681 TI - [Biological and health effects on electric and magnetic fields at extremely low frequencies]. AB - While the enormous benefits of using electricity in everyday life and health care are unquestioned, during the past 20 years the general public has become increasingly concerned about potential adverse health effects of exposure to electric and magnetic fields at extremely low frequencies (ELF). Such exposures arise mainly from the transmission and use of electrical energy at the power frequencies of 50/60 Hz. The international scientific community is addressing the associated health issues through in vitro, in vivo and epidemiological studies, although present research results are often contradictory. The purpose of this paper is to provide information about the possible impacts of exposure to electric and magnetic field at ELF frequencies on health within the community and the workplace. Information comes from recent research studies on this subject and from epidemiological pooled studies recently reviewed by eminent authorities. PMID- 16353682 TI - [Poverty and nutrition: a survey on the under age sons of the beneficiaries of the R.M.I. in the L'Aquila District Area]. AB - The alimentary education, let alone the relative behaviour, represent an important preliminary statement for a children perfect growth. The results of a not healthy nutrition or alimentary mistakes can produce several problems. The study aims to analyze the child and adolescent nutrition for families with economic difficulties that receive a (social and economic) benefit called "Reddito Minimo di Inserimento" (RMI), within the L'Aquila district area. For the valuation of nutrition state, it has been used the BMI (Body Mass Index), with the necessary corrections for sex and age. From the made analysis, the children's about 33% is classifiable as "at risk of soprappeso" or as "soprappeso", with prevalence for the males, above all in the classes of younger ages, like the most recent studies on the children's nutrition. PMID- 16353683 TI - [Cremation plant planning and related hygienic-sanitary aspects]. AB - The number of cremators is set to grow in Italy owing to current increasing demand for cremation, as a system of burial as well as a system of disposal of non-decomposed human remains resulting from operations of exhumation (as provided by the Circular No. 10 of 31 July of 1998 of the Ministry of Health). Some indications on how town planners need to size up the cremation plant with regard to its capacity to face cremation demand, in order to avoid any hygienic-sanitary inconveniences are given. As a matter of fact, for planning the demand for cremation from Circular No. 10 of 31 July of 1998 it is necessary to assess the decomposition time in burial sites. PMID- 16353684 TI - WHO Expert Committee on Specifications for Pharmaceutical Preparations. AB - This report presents the recommendations of an international group of experts convened by the World Health Organization to consider matters concerning the quality assurance of pharmaceuticals and specifications for drug substances and dosage forms. Of particular relevance to drug regulatory authorities and pharmaceutical manufacturers, this report discusses the monographs on antiretrovirals proposed for inclusion in The International Pharmacopoeia and specifications for radiopharmaceuticals, quality specifications for antituberculosis drugs and the revision of the monograph on artemisinin derivatives, as well as quality control of reference materials, good manufacturing practices (GMP), inspection, distribution and trade and other aspects of quality assurance of pharmaceuticals, and regulatory issues. The report is complemented by a number of annexes, including an amendment to good manufacturing practices: main principles regarding the requirement for the sampling of starting materials, guidelines on good manufacturing practices regarding water for pharmaceutical use, guidelines on the sampling of pharmaceutical products and related materials and draft guidelines for registration of fixed-dose combination medicinal products. PMID- 16353685 TI - [The 3' by 5' WHO/UNAIDS initiative--can 3 million people obtain HIV therapies by the end of 2005]. PMID- 16353686 TI - [Gastric carcinoma (adenocarcinoma of the stomach). Main symptoms: abdominal pain, weight loss, iron deficiency anemia]. PMID- 16353687 TI - [Gastroesophageal reflux disease]. AB - Gastroesophageal reflux disease (GERD) includes different manifestations of esophageal and extraesophageal affections, such as erosive reflux disease, Barrett's esophagus, laryngitis and even pulmonary diseases. Typical symptoms are heartburn and acid regurgitation, but there are a lot of atypical disturbances like hoarseness or chronic cough. Endoscopy is the most important diagnostic procedure for elderly patients and for those with atypical symptoms. Younger people without alarming symptoms can be tested for GERD by treatment with proton pump inhibitors (PPI). The therapeutic approach depends on the endoscopic findings. PPI are preferred drugs to manage most GERD-related problems. Anti reflux surgery is appropriate for selected patients. Long-term followup data will help to define the benefit of new endoscopic procedures. Barrett's esophagus has to be regarded as a precancerous lesion, therefore surveillance strategy is required. PMID- 16353688 TI - [Drug therapy in chronic liver disease]. AB - Treatment of virally induced chronic hepatitis consists of interferons, nucleotide and nucleoside analogues. So far, combination therapies are not superior to a monotherapeutic regimen. If detected early, inherited metabolic diseases (e.g. hereditary hemochromatosis, M. Wilson) can be successefuly treated, leading to a normal life expectancy of the patients. In contrary treatment of cholestatic chronic liver disease, especially of PSC, is far from being satisfying, due to our very limited knowledge of the etiology of this diseases. The same holds true for the therapy of NASH. Weight reduction and treatment of the often underlying diabetes are therapeutic cornerstones in the moment. On the other hand, immuosuppression, some times life-long, remains a (highly successful) treatment option in cases of proven autoimmune-hepatitis. PMID- 16353689 TI - [Gastroenterology in the elderly]. AB - The medical care of old patients becomes more and more important. Currently about 21% of all people are older than 60 years. Decisions about therapeutical procedures often cannot be based on guidelines, because studies leading to this guidelines have normally excluded older patients. Decisions have often to be made individually in view of different factors like comorbidity and social structures. All symptoms occuring in older patients need a careful diagnostic work-up, even the majority of complaints as in younger people are of functional origin. In view of the high life expectancy and quality of life the use of difficult and expansive diagnostic and therapeutical tools are justified also in older people. So diagnostic and interventional endoscopic procedures can be done with low risk in older patients. PMID- 16353690 TI - [Coma]. PMID- 16353691 TI - [Psychiatrics emergencies]. PMID- 16353692 TI - Management of pain in children: a review of the literature. AB - BACKGROUND: Pain is a very common situation encountered in clinical practice. It is as common in children as in adults and often it is underevaluated and undertreated. The aim of this review is to stimulate and sensitize paediatricians and other child health care providers to the importance and necessity of adequate appreciation, evaluation, and management of pain in children. METHODS: Review of the pertinent literature. Selected references to highlight the mechanism of pain, pain assessment tools and therapeutic approach to management, were included in this review. RESULTS: Pain in children is underevaluated and undertreated. This is so even in developed countries. A study put the prevalence of pain among children at 26%, and that two out of every three patients with pain were not detected, and that one out of three patients detected received no treatment. Popular dogma held that the human child does not feel pain and that giving the child powerful analgesics may lead to addiction. Despite the availability of several pain assessment tools, they are under used in paediatric practice. Pain pathways develop in the second trimester, so the smallest neonate is able to respond to noxious stimulation. CONCLUSION: Childhood pain is still underevaluated and under treated. The adequate management of pain in children is an important factor in better outcome of childhood illnesses. PMID- 16353693 TI - Molecular biology and medicine: a review of developments. AB - BACKGROUND: The human genome-sequencing project was completed in April 2003, in the same year that marks the 50th anniversary of the discovery of the double helical structure of DNA. Much of the knowledge derived from the huge number of discoveries in molecular biology research over the past five decades and the genome project has the potential to be of immense benefit world-wide. Such knowledge is already of widespread application in the developed world and much more potential in the future. METHODS & RESULTS: This article briefly reviews the literature of research works on molecular biology in English language and discusses some of the findings and highlights recent developments and future trends. CONCLUSIONS: The advancement in molecular biology presents huge opportunities and potentials for improved health services worldwide. The article also proposes an approach whereby benefits could be reaped from the application of molecular biology techniques and the information derived from the human genome project in sub-Saharan Africa. PMID- 16353694 TI - Structural heart diseases in Nigerian children. AB - BACKGROUND: Cardiovascular diseases both in adults and children constitute a major public health problem and structural heart diseases are an important group of disorders in children worldwide. The pattern of this group of disorders however, varies between regions and countries and even within countries. Recognizing the structural cardiac conditions that prevail in a particular area is important in health planning and for improving health care services. The present survey sets out to describe the pattern of structural heart diseases among children in Aminu Kano Teaching Hospital, Kano from our echocardiography data. METHODS: The echocardiographic reports of all children seen in Aminu Kano Teaching Hospital, Kano between August 2002 and September 2004 (24 months) were reviewed. Information obtained from the records includes age, gender, clinical diagnosis and echocardiographic findings. Data was analyzed using SPSS version 10.0 software. RESULTS: A total of 108 children, aged between two weeks and eighteen years, were referred for echocardiographic examination in the 2-year study period. Of these, 88 had an abnormal echocardiogram. There were 55 boys and 31 girls, giving a male and female ratio of 1.8:1. Congenital heart diseases accounted for 55 (62.5%) of the studied subjects while acquired heart diseases were responsible for 33 (37.5%). Isolated ventricular septal defect (VSD) was the commonest congenital heart disease. Rheumatic valvular heart diseases were the commonest acquired structural heart disease. CONCLUSION: With the establishment of tertiary healthcare institutions in Nigeria, availability of echocardiographic facilities as well as increasing number of paediatric cardiologists, more of these cases are likely to be seen in the future. There is an urgent need for the government to establish a well equipped cardiothoracic surgical centre to cater for these patients either free or at highly subsidized rates. PMID- 16353695 TI - Pattern of morbidity among pre-school children attending the children's outpatient clinic of Federal Medical Centre Owerri, Nigeria. AB - BACKGROUND: Several interventional programmes which sought broadly to promote the health of the child and also prevent diseases were introduced globally in the latter period of the last century. This study set to describe the pattern of morbidity among pre-school children attending the children's outpatient clinic of the Federal Medical Centre, Owerri. METHOD: The case records of pre-school children who presented at the clinic between January and April 2004 were collected and entered into a spread sheet. The age, sex and diagnosis made were extracted from the records. In cases where more than two diagnoses were made, the most likely two following a review of presenting complaints and findings on examination were chosen. RESULT: Eight hundred and twenty nine patients consisting of 468 (56.2%) males and 361 (43.5%) females met the inclusion criteria. Patients aged 0-11 months constituted 47.8% while those between 48 and 59 months accounted for 1%. The five commonest causes of morbidity were malaria (60.7%), acute respiratory infection (35.8%), diarrhoeal disease (7.4%), skin infection (6.8%) and urinary tract infection (3.6%). The prevalence of malaria and acute respiratory disease were highest in patients aged between 12-23 months while diarrhoea was highest in the group 0-11 months. The prevalence of vaccine preventable diseases such as measles and tuberculosis were low accounting for 1.1% and 1% of morbidity respectively. CONCLUSION: This study shows that morbidity from vaccine preventable diseases is low, which is likely a reflection of the increased emphasis on immunization programmes. The major causes of morbidity in pre-school children in Owerri are still common diseases that have been around for a while and are basically preventable. PMID- 16353696 TI - Tuberculosis in Calabar: a ten-year review (1994-2003). AB - BACKGROUND: The aim of the study is to determine the pattern of tuberculosis as seen in Calabar over 10 years (1994-2003). METHODS: A ten-year retrospective study (1994-2003) was carried out to evaluate the pattern of tuberculosis among subjects in Calabar, Nigeria. A total of 6,737 cases of confirmed pulmonary and extrapulmonary tuberculosis seen during the period of survey were reviewed. The cases were drawn from the Infectious Disease hospital (IDH) Calabar, University of Calabar teaching hospital (UCTH), and TBL unit of the Ministry of Health, Calabar. Subjects, apart from being symptomatic were confirmed to have Pulmonary Tuberculosis (PTB) by Ziehl-Neelsen's sputum staining technique. Positive confirmation was attained if a patient's sputum was positive for acid fast bacilli on at least two separate times in line with the World Health Organisation's (WHO's) recommendation. In addition, some patients had chest radiograph, and those with evidence of pleural effusion were confirmed by pleural aspirate. However, cases with extrapulmonary tuberculosis (EPTB) were considered separately and the site of the lesions noted. RESULTS: The results revealed some progressive yearly increase in the number of tuberculosis [TB] cases over the 10 year period with more children being diagnosed. Tuberculosis of the spine was the most common type of extrapulmonary tuberculosis among the patients with EPTB. The treatment outcome over the period was not satisfactory with a cure rate of 57% and a mean mortality rate of 14% (with a range of 12% to 17%), and a default rate of 18%. CONCLUSION: The implication is that either the available tools are not properly utilized or extra-measures will be required to contain the scourge. We therefore recommend an increased supervision of the intensive phase of therapy and provision of facilities for culture and drug sensitivity testing at treatment centres, in case drug resistance is a factor. PMID- 16353697 TI - Conservative management of tuberculous spondylitis in a developing country. AB - BACKGROUND: Tuberculous spondylitis is a significant health burden in the Third world. Tuberculosis and its complications are on the increase because of the HIV/AIDS challenge. The aim of this study was to review the patients managed for tuberculous spondylitis at the University of Port Harcourt Teaching Hospital, Port Harcourt, Nigeria. METHODS: This was a retrospective study of patients seen at the University of Port Harcourt Teaching Hospital between January 1999 and December 2002 with tuberculosis of the spine. Patients case notes were reviewed after collating out-and-in-patient records. Analysis was done using multiway frequency tables. RESULTS: There were 16 males and 18 females, with a ratio of 1:1.1. Their ages ranged from 3.3 to 82 years with an average of 31.1. Peak ages of occurrence were the 3rd and 4th decades. Eighteen patients (52.9%) presented within the first 6 months of symptoms. The most frequent presenting symptoms were back pain, weight loss, difficulty with walking, low grade fever, and night sweats. The most frequent signs were gibbus and lower limb paralysis. The most common association was pulmonary tuberculous (32.4%). The thoracic spine was mostly affected, followed by the lumbar spine. Wedge collapse of the vertebrae was the common radiological finding. Twenty two patients had multilevel affection. Twenty patients (71%) had a lymphocyte differential of more than 45%, 87% had erythrocyte sedimentation rate of over 20 mm/hr. Westergren, and Mantoux test was positive in 55 percent of the patients. Twelve patients had outpatient treatment and 22 had in-patient treatment. Duration of hospital stay ranged from 7 to 157 days with an average of 62.6 days. All the patients had standard combination antituberculosis therapy. No patient had surgery for tuberculous spondylitis. Outcome of treatment was good (68.2%). Follow up was poor with a high default rate. There were 3 mortalities (8.8%). CONCLUSION: Diagnostic delays partly due to lack of experience makes a high index of suspicion necessary to make a diagnosis. Outcome utilizing conservative methods of treatment is good. PMID- 16353698 TI - Blindness and low vision in adults in Ozoro, a rural community in Delta State, Nigeria. AB - BACKGROUND: The objective of this article is to determine the prevalence and causes of blindness and low vision in Ozoro, a rural town of Delta State in Nigeria, in order to provide baseline information for planning a prevention of blindness programme. METHODS: A cross sectional study using a stratified random sampling method was used to select the compounds. The five communities in the town formed the different sub-groups or strata. Visual acuity were recorded for all adults 40 years and older. Persons who had visual acuity less than 6/18 were further examined to determine the cause of low vision or blindness. The WHO definition of visual impairment according to visual acuity was used as criteria for classification. RESULTS: A total of 815 persons (coverage of 51.7%) were examined using a modified WHO PBL record of blindness form. The prevalence of blindness (VA < 3/60 in the better eye) for people of 40 years and above was 6.3% (95% CI, 4.6% to 8%) and low vision VA 6/24 to 3/60 in the better eye was 25.2%. The estimated prevalence of bilateral blindness for all ages was 1.3% and low vision was 5%. The rate of blindness and low vision increased with age being highest in subjects who were 60 years and above. The main causes of blindness and low vision were cataract, accounting for 60% of all bilateral blindness and 51.7% of all low vision. Other causes of blindness include, posterior segment diseases (11.7%). Glaucoma was presumed to be cause of blindness in 9.8% of cases; others were uncorrected aphakia 5.9%, and globe abnormalities 5.9%. Refractive error was the second major cause of low vision accounting for 22% of bilateral low vision. CONCLUSION: The magnitude of blindness and low vision in this oil rich Ozoro community in Delta State is high and majority are avoidable causes of blindness. PMID- 16353699 TI - Urethral prolapse: a retrospective analysis of hospitalized cases in Port Harcourt. AB - BACKGROUND: Urethral prolapse is a disease of the prepubertal girls and postmenopausal women. It is rare and seen in most cases as a circular protrusion of the distal urethra through the external meatus. METHOD: A retrospective study of cases managed in University of Port Harcourt Teaching Hospital (UPTH) between January 1989 and December 2000 were analysed. RESULTS: Nineteen (0.4%) excisions of urethral mucosal prolapse out of a total of 4351 gynaecological surgeries were carried out between January 1989 and December 2000 in UPTH. Of the 35 total paediatric gynaecological admissions, there were 17 (49%) cases of urethral prolapse. It occurred commonly between ages 4 and 7 years (63%). Common presenting complaints included vaginal bleeding, vaginal discharge and mass protruding from the urethral opening. Excision was found curative in all cases. CONCLUSION: There is a place for promotion of female education and community sensitization to encourage early presentation and treatment. This will reduce infectious morbidity and its sequelae in our environment. PMID- 16353700 TI - Management of cervical incompetence in Aba, south-eastern Nigeria. AB - BACKGROUND: There have been relatively few reports on the outcome of cervical cerclage performed in non-tertiary centres. The aim of this study was to determine the pregnancy complications and outcome in patients following cervical cerclage. METHODS: Seventy-one patients who had 103 pregnancies and underwent cervical cerclage at Women's Hospital, Aba over a ten-year period were reviewed. The diagnosis of cervical incompetence was made from the history in 90.6% of cases and also in some cases by hysterosalpingography and ultrasonography. RESULTS: The Shirodkar (65%) and McDonald (35%) techniques were employed and 10.7% of cases were done as emergency procedures. The more common post-insertion complications were urinary tract infection (50.4%), preterm rupture of membranes (20.4%) and vulvovaginitis (14.6%). Common labour associated complications were antepartum haemorrhage, perineal/cervical tears and malpresentations. The preterm birth rate was 32%. Term births accounted for 68% of the deliveries. The overall fetal salvage rate was 92.2%. Antepartum haemorrhge was a significant indication for caesarean section delivery. The perinatal mortality was 63.2 per 1000 and there was no maternal death. CONCLUSION: The high fetal salvage rate of 92.2% justifies the procedure of cervical cerclage. PMID- 16353701 TI - Spousal communication in contraceptive decisions among antenatal patients in Calabar, south eastern Nigeria. AB - BACKGROUND: The objective of this study was to determine how many women in Calabar urban community actually discuss with their spouses about family planning, how their spouses view family planning and how that affects their usage of family planning facilities. METHOD: The study was conducted using a structured questionnaire which was administered by house officers and medical students on one thousand consecutive patients who attended antenatal care during the study period Seven hundred patients met the inclusion criteria for the study. RESULT: Fifty six percent of the respondents in the study have never discussed family planning with their spouses. About 15% of the respondents have the same view on family planning with their husbands while 11% of the women themselves do not want family planning even though their husbands wanted it. CONCLUSION: Communication between couples about family planning should be encouraged. Whereas women have common meeting points like antenatal clinics to learn more about contraceptives and family planning, the men rarely have such opportunities. They will therefore benefit from discussing with their spouses who are opportuned to learn from the various clinics. Spousal communication should therefore be emphasized in these clinics where contraception and family planning are taught. PMID- 16353702 TI - Five-year experience with depot medroxy progesterone acetate injectable contraception. AB - BACKGROUND: The objective of this article is to describe the characteristics and experiences of clients who used depot medroxy progesterone acetate for contraception. METHODS: A retrospective review of family planning records of clients who used depo-provera between 1992 and 1996 at the family planning clinic of University of Nigerian Teaching Hospital, Enugu. RESULTS: The mean age of clients was 32.7 +/- 5.2 years. Over 71% of clients had five or more children. Clients with no education (78.7%) or primary education (75.1%) were more likely to be grand-multiparous compared to women with at least secondary education (49.6%). Among clients with 1-4 living children, 81.3% used depo-provera for child spacing. Among those with 5-8 living children, 48.8% used it for child spacing. Menstrual abnormalities were the main complications but the commonest reason given for discontinuation was "husband's wish". The continuation rate at 36 months was 82.7%. CONCLUSION: Depot medroxy progesterone acetate is an acceptable and effective method of contraception. The involvement of men in family planning programmes cannot be over-emphasised as they significantly influence the decision of women regarding contraceptive use. PMID- 16353703 TI - Spectrum of otorhinolaryngology emergencies in the elderly in Ibadan, Nigeria. AB - BACKGROUND: The elderly (65 years and above) are a special group of patients whose illnesses and management are modified by the process of aging. It is thus the aim of this study to evaluate the spectrum of ear, nose, throat, head and neck emergencies among the elderly in Ibadan, Nigeria. METHOD: This is an eight year (April 1996-March 2004) retrospective study of 87 geriatric patients who presented with otorhinolaryngological emergencies at our centre. RESULTS: Eighty seven emergency cases studied constituted about 20% of the total geriatric cases seen within this period. There were 55 (63.2 per cent) males and 32 (36.8 per cent) females. The pharyngolaryngeal emergencies with upper airway obstruction constituted 55 per cent of the cases, a majority being a result of malignant lesions of the larynx and pharynx. Others were nasal 20.7 per cent, otological 11.5 per cent, oesophageal 9.7 per cent and head and neck 3.1 per cent. Most of the patients had emergency tracheostomy (54 per cent), as a prelude to their definitive management while 24 per cent were managed medically. CONCLUSION: Prevalence of emergency otorhinolaryngology cases among the elderly appears high. Most of the throat diseases with upper airway obstruction were due to malignancy. There is thus the need for increased awareness of the people through social campaigns and health education on the merits of early detection and seeking appropriate treatment for these tumours. PMID- 16353704 TI - Retropharyngeal abscess: a clinical experience at the University College Hospital Ibadan. AB - BACKGROUND: Infection within the retropharyngeal space could progress on to an abscess formation resulting into retropharyngeal abscess (RPA), which can either be acute or chronic. RPA can be a life-threatening emergency, with potential for airway compromise and other catastrophic complications. This review is aimed at highlighting our experience with patients with a diagnosis of retropharyngeal abscess. METHODS: Retrospective review of thirty patients with a confirmed diagnosis of retropharyngeal abscess, who were admitted and managed within an eleven-year period (1993 to 2003) in the Otolaryngology department of the University College Hospital Ibadan. RESULTS: There were fifteen males and females each with M:F ratio of 1:1, consisting of twenty-five children and five adults and their median age was 21 months and twenty three (77%) were younger than 5 years. It was observed that while the adult patients presented early for specialist treatment, the paediatric patients presented late. The major complaints were fever (87%), respiratory distress or stridor (57%), cough (53%), neck pain/swelling (43%), and refusal of feeds (30%). Other minor complaints were throat pain, difficulty in swallowing, anorexia, and weight loss. The commonest associated symptoms seen especially among the younger age group were nasal discharge, nasal blockage, tooth-ache, snoring and limitation of neck movement. CONCLUSION: The treatment of retropharyngeal space infections in children and adults should include accurate clinical diagnosis, empirical usage of broad spectrum antibiotics, and timely surgical drainage. PMID- 16353705 TI - Seroprevalence of HIV among unemployed individuals undergoing pre-employment medical examination in Port Harcourt, Nigeria. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection is highly endemic in Nigeria, particularly with the prevalence in 2001 at 5.8%. This study was undertaken to determine the prevalence of HIV among unemployed individuals undergoing pre-employment medical examination in Port Harcourt. METHOD: HIV screening was performed on 868 individuals comprising 373 males and 495 females presenting to the University of Port Harcourt for the purpose of pre-employment medical examination using a double ELISA confirmatory test of Immunocomb and Genscreen HIV 1 & 2 kits. RESULTS: The sero-prevalence rate was 27/868 (3.19%) among the total population. HIV seroprevalence was relatively higher among females 18/495 (3.6%) compared to males 9/373 (2.4%). The highest prevalence was found in the <19 years age group 7/135 (5.1%) and lowest in the 40-49 years age group 3/130 (2.3%), although the difference was not statistically significant (chi2 = 4.86, p = 0. 09). The highest prevalence occurred among separated subjects 2/26 (7.7%) compared to singles 18/460 (3.9%) and married subjects 7/382 (1.8%). CONCLUSIONS: This study indicates a 3.1% prevalence of HIV infection among unemployed individuals studied and calls for urgent and concerted efforts aimed at promoting behavioural, cultural and social changes that will reverse the current trend in the prevalence of HIV among Nigerians. PMID- 16353706 TI - Bacteriology of urinary tract infection among patients with acquired immunodeficiency syndrome in Jos, Nigeria. AB - BACKGROUND: The study was set to find out the bacterial agents causing urinary tract infection (UTI) in acquired immunodeficiency syndrome (AIDS) patients. METHODS: Two hundred patients with AIDS on admission at the Jos University Teaching Hospital (JUTH) were recruited into this study. Urine sample was collected from each patient and processed. Other information was obtained with protocol questionnaire. The results were analyzed using Epi Info 6. RESULTS: The prevalence of UTI in AIDS patients was found to be 24% and control 10.6%. Escherichia coli were the commonest Gram-negative isolate followed by Klebsiella species. Enterococcus species was the commonest Gram-positive organism. CONCLUSION: There should be extreme caution whenever the need arises to catheterize AIDS patients. PMID- 16353707 TI - Hepatitis B virus infection in a rural settlement of northern Nigeria. AB - BACKGROUND: Hepatitis B virus (HBV) infection has been present in our communities for quite sometime now. However, majority of the people especially in rual areas are not aware of its devastating effects as adjudged by their socio-cultural practices. METHODS: Three hundred randomly selected residents of Zawan community (comprising 193 males and 107 females) were screened for hepatitis B surface antigen (HBsAg) presence in their sera. Other information was obtained with the aid of a structured questionnaire and results analysed using Epi Info 6 statistical software. RESULTS: The prevalence of HBV in Zawan village was found to be 12.6% (9% among males and 3.6% among females). The risk factors associated with HBV transmission were tonsillectomy, tattooing, use of sharp objects as well as trading. Marital status was also found to be a significant risk factor as there was a high prevalence of HBV infection among the widowed/divorced/separated group. CONCLUSION: HBV infection is endemic in our rural communities just as it is in the urban areas and efforts should be made to step up its health education campaign as well as the interventional mechanisms including immunization. PMID- 16353708 TI - Necrotizing fasciitis of the external genitalia following traditional circumcision. AB - BACKGROUND: Necrotizing fasciitis of the external genitalia is not common in children, and is particularly unusual after circumcision. METHOD: A case report of necrotizing fasciitis of the external genitalia in a one month old boy with discussion of relevant literature. RESULT: A one-month-old boy presented with necrotizing fasciitis of the external genitalia following traditional circumcision. Treatment consisted of early administration of intravenous broad spectrum antibiotics and debridement. Skin grafting was not necessary as the wound contracted and healed by secondary intention with minimal scarring. CONCLUSION: Early institution of appropriate antibiotics and debridement should prevent morbidity and mortality from this uncommon condition. PMID- 16353709 TI - Urethral prolapse in a woman in her reproductive age--a very rare occurrence. AB - BACKGROUND: Urethral prolapse is a rare condition occurring in prepubertal girls and postmenopausal women. Even most rare is that occurring in the reproductive age grade. METHOD: A report of a woman in her reproductive age who presented in the gynaecology clinic of a private hospital is presented together with a brief review of the literature. RESULTS: Urethral prolapse associated with uterovaginal prolapse, lower abdominal pain and dysuria in a 48-year old grand multiparous (para 8 + 0) farmer who is still observing her monthly menstrual cycle was seen. Her pregnancies and deliveries were unsupervised. A urine culture revealed moderate growth of Escherichia coli sensitive to ciprofloxacin. The urethral prolapse resolved satisfactorily on this. CONCLUSION: It is advocated that antenatal care, education and women empowerment be made a priority. This will almost eradicate this very rare condition in our society. PMID- 16353710 TI - Congenital inclusion cyst of the subgaleal space: a case report. AB - BACKGROUND: Subgaleal cysts are epidermal cysts located under the galea aponeurotica. They are rare lesions but are commoner among the children of African descent. To the authors knowledge, none has been described in Kano, Northwestern, Nigeria. METHODS: We present the management of a 6 month old Nigerian boy with scalp swelling. RESULT: The scalp swelling was noticed soon after birth and was progressively increasing in size. There was no preceding fever or trauma to the scalp and no associated pain. At six months, the child could sit without support and had attained adequate neck control. Examination revealed a swelling over the anterior fontanelle, measuring 3x2cm. The swelling was hemispherical, firm, fluctuant, and non-tender. It was not pulsatile. Its size did not increase on crying. Diagnosis was confirmed by plain x-ray film and transfontanelle sonography. Total excision of the mass was carried out under general anesthesia and histology confirmed the diagnosis of epidermal inclusion cyst. The child was discharged home in good condition. CONCLUSION: Awareness of this lesion will avoid misdiagnosis and inappropriate treatment. PMID- 16353711 TI - Corrosive-induced gastric outlet obstruction without oesphageal involvement: a case report. AB - BACKGROUND: The objective of this paper is to report an unusual case of isolated gastric outlet obstruction following corrosive ingestion. METHOD: A case report of a 28-year old female seen by the authors. The literature on gastric outlet obstruction following ingestion of corrosives is reviewed briefly. RESULTS: Features of worsening gastric outlet obstruction were found in this 28-year old female five months after ingestion of hydrochloric acid. There was an antecedent history of depressive illness. The upper gastrointestinal barium contrast radiographs showed a normal oesophagus and proximal stomach. The distal stomach was however scarred, contracted with severe antropyloric stenosis. She underwent nutritional rehabilitation with high protein diet and made an uneventful recovery after a gastrojejunostomy. CONCLUSION: This case suggests a relative resistance of the oesophagus to corrosive acids as reported in the literature. The stomach, however, is more susceptible to acids causing burns with subsequent cicatrisation around the antrum and pylorus. PMID- 16353712 TI - Electrocution attributed to supernatural forces in the Niger delta region of Nigeria: a report of three cases. AB - BACKGROUND: Accidental deaths from electrocution in this environment are sometimes related to charm and witchcraft caused by one's enemies. The aim of this article is to highlight the problems of beliefs and accidental electrocution. METHOD: We report three cases of accidental electrocution which was believed to be associated with traditional beliefs of witchcraft and charm. RESULTS: Autopsy findings in two cases showed characteristic electric burns (the joule burn which is the area of entry) without obvious organ changes. The third case revealed no burn but investigation of the scene of incidence and other sources of information are consistent with a diagnosis of electrocution. CONCLUSION: Post mortem examination can assist to dispel misconception and unnecessary belief on cause of death in our community. PMID- 16353713 TI - Megaloblastic anaemia: response to Amples A and B (folic acid, vitamin B12 (Cyanocobalamin), niacin and vitamin C)--a case report. AB - BACKGROUND: Megaloblastic anaemia is prevalent in our society and patients are diagnosed late. METHOD: Case Report of a patient with megaloblastic anaemia and discussion of relevant literature. RESULTS: A 50 year old Nigerian trader with recurrent chronic anaemia and repeated blood transfusions (Eight units in 10 days) was finally diagnosed to have megaloblastic anaemia. He was commenced on intramuscular, Ample A and Ample B (folic acid 0.7mg, vitamin B12 (Cyanocobalamin) 2500g, niacin 12mg and vitamin C 150mg) on alternate days (6 doses) then weekly as maintenance until haematocrit returned to normal. Response was dramatic; haemoglobin was 6.4g/dl before, 7.9g/dl after the second dose and 11.5g/dl after the 6th dose of the drug. The corrected retic was 2.0% before, 4.6% 72hrs after and 8.4% after the 6th dose. The MCV decreased from 105fl before to 79fl after the 6th dose of the drug. His packed cell volume has remained above 35% after completing the 6th dose and during maintenance therapy. CONCLUSION: The combination of Cyanocobalamin, folic acid, niacin, and vitamin C, in Amples A and B is complementary. Treatment with Amples A and B is effective and affordable in the management of megaloblastic anaemia. PMID- 16353714 TI - Fatal relapse of hyperreactive malarial splenomegaly (HMS) in a 10-year old Nigerian female--a case report. AB - BACKGROUND: The Hyperreactive Malarial Splenomegaly Syndrome (HMS) originally called the tropical splenomegaly syndrome (TSS) or Big spleen disease refers to cases of splenomegaly in the tropics for which no cause was found despite thorough investigation. It is restricted to the malarial belt, yet there are few reports on HMS in Nigeria, probably due to a low index of suspicion and non availability of laboratory facilities to determine titres of malarial antibodies. The objective of this paper is to highlight the features, management, risk of relapse and prognosis of HMS. METHOD/RESULT: We present a 10-year old female with recurrent massive splenomegaly with previous clinical response to antimalarials and evidence of hypersplenism. CONCLUSION: HMS should be suspected in any child with moderate to massive splenomegaly with evidence of hypersplenism and clinical response to antimalarials. PMID- 16353715 TI - Clinical photographic quiz. PMID- 16353716 TI - Deliveries during a nurses' strike in Port Harcourt. PMID- 16353717 TI - Dealing with diabetes in the school setting--update on diabetes resources. PMID- 16353718 TI - Combating childhood tooth decay. Avoiding the sugar that lurks in cafeteria lines and lunchbox favorites. PMID- 16353719 TI - Pediatric weight issues: what are healthcare providers and parents to do? PMID- 16353720 TI - School Nurse News Recognition Award. PMID- 16353721 TI - The perils of peanuts. PMID- 16353723 TI - Making our lives count: youth in partnership with health professionals. PMID- 16353722 TI - School health report card. AB - School nurses all across the nation perform valuable screenings every year for the students in their schools. These screenings identify abnormalities that have the potential to affect their education. With skill and determination, a variety of health screenings are performed and the results communicated with the parents of the children. A health "report card" is a proposed method of communicating vital health screening information, including the height, weight, and BMI of every student. This is directly related to the epidemic of childhood obesity. Current life styles promote minimal exercise and increased consumption of food that has minor nutritional value. It is time to identify affected children and become engaged in resolving this health crisis, by teaching this nation's children behaviors that will result in optimal health. PMID- 16353724 TI - Similarities stronger than differences. Career & salary survey 2005. PMID- 16353725 TI - Re: career & salary survey, March 2005 issue. PMID- 16353726 TI - Think before you speak. PMID- 16353727 TI - New research on late abortion. PMID- 16353728 TI - Bottle-feeding: report shows pressure on mothers. PMID- 16353729 TI - RCM conference. Motherhood and the mind. PMID- 16353730 TI - A new beginning. PMID- 16353731 TI - Policy into practice. PMID- 16353732 TI - Postpartum psychiatric illness: can women recognise the onset? AB - In the Department of Health's Confidential Enquiry into Maternal and Child Health (2004), deaths from psychiatric causes are reported to be the most common cause of maternal death overall. Midwives are in a prime position to inform women regarding this issue. This paper explores the lack of client understanding of postnatal depression through a small, personal exploratory study of a community midwife's routine postnatal visits. PMID- 16353733 TI - Rebirthing midwifery. PMID- 16353734 TI - Expanded newborn screening: anticipating the future. PMID- 16353735 TI - Inverting the hierarchy. PMID- 16353736 TI - Litigation and resolution. PMID- 16353737 TI - New on-call system for the community. PMID- 16353738 TI - Midwives: professional servants? PMID- 16353739 TI - Midwives: professional servants? PMID- 16353740 TI - Society's most vulnerable get a helping hand. PMID- 16353741 TI - Are students too academic to actually care? PMID- 16353742 TI - Maintaining confidentiality: for whose benefit? Rogers v. NYU Hospital Center. PMID- 16353743 TI - [The third year]. PMID- 16353744 TI - [Metronomic chemotherapy. Should we change our thinking in the treatment of advanced tumors?]. PMID- 16353745 TI - Skin diseases described in Japan 2004. AB - During the last century of modern dermatology, more than 30 skin diseases have been described first by physicians from Japan. Many of those conditions were disorders of pigmentation and keratinization, which are quite common in Oriental patients. Since the late 1940s, a number of skin diseases first reported in Japan have gained attention internationally among them being Kimura disease, hypomelanosis of Ito, Kawasaki disease, adult T-cell leukemia/ lymphoma, eosinophilic pustular folliculitis, prurigo pigmentosa, and Ofuji's papuloerythroderma. In this article, we review skin diseases that were first established as distinct entities in Japan, in order to familiarize readers of the Western literature with these conditions. PMID- 16353746 TI - Metronomic low-dose chemotherapy as antiangiogenic therapeutic strategy for cancer. AB - New blood vessel formation is essential for the growth and metastasis of many cancers. As a result, antitumor activities of various angiogenesis inhibitors have been intensely explored in various tumors. Recent preclinical studies suggest that certain conventional cytotoxic agents can function as antiangiogenic drugs when administered at comparatively low doses on a continuous or very frequent schedule. Such antiangiogenic 'metronomic' scheduling of chemotherapy without extended rest periods has been shown to exert significant therapeutic antitumor efficacy with very limited toxicity in different tumor models. Combining metronomic low-dose chemotherapy regimens with specific angiogenesis inhibitors further increases efficacy. Based on the promising preclinical studies, it is anticipated that metronomic chemotherapy in combination with angiogenesis inhibitors will prove effective in clinical trials in terms of survival prolongation. While considerable progress may derive from larger randomized clinical studies, only joint efforts between basic and clinical research will ultimately advance the new paradigm of long-term metronomic antiangiogenic chemotherapy, which carries the prospect of turning cancer into a more controllable chronic disease at minimal toxicity. PMID- 16353747 TI - [Skin protection and skin disease prevention courses for secondary prevention in health care workers: first results after two years of implementation]. AB - BACKGROUND: Healthcare workers have an increased risk of occupational dermatoses. In January 2002, the Department of Social Medicine, Occupational and Environmental Dermatology of the University of Heidelberg started organizing special prevention courses for this group of employees in cooperation with the Accident Prevention & Insurance Association for Health Care Workers (BGW). The major aims are to improve individual skin protection and skin care habits, as well as to optimize diagnostic procedures and therapy. PATIENTS AND METHODS: The two-day course has up to 14 participants. Teaching units mainly focus on skin structure and function, general aspects of occupational skin diseases, general information concerning skin protection and practical exercises emphasizing the correct use of skin cleansing and skin protection products. Additionally, every participant undergoes a dermatological examination including a detailed history and skin inspection. Individual skin protection strategies are developed, and the participants are provided with the opportunity to ask the dermatologist questions in confidence. RESULTS: Most of the 355 participants have been female (87.3%) with a mean age of 36.9 years. 95% had hand eczema, predominantly dyshidrotic morphology. In many cases, there was a mixture of atopic, irritant and allergic contact dermatitis but irritant contact dermatitis was most common (43%). Atopy was present in 68% of the patients. The participants rated the course as good to excellent. Other benefits are the exchange of experience between patients working in the same or similar occupations and transfer of the new knowledge to colleagues, family and friends. CONCLUSIONS: Examinations and advisory services in occupational dermatology are still fragmentary. Skin barrier creams and moisturizers are not sufficiently utilized in daily practice. In the future, similar courses should be offered for employees in other professions with an increased risk of occupational skin diseases. PMID- 16353748 TI - EMPACT syndrome. AB - BACKGROUND: Seizure prophylaxis with phenytoin is a common measure in oncologic patients with brain metastases. In these patients, generalized severe adverse drug reactions such as erythema multiforme (EEM) may occur. However, in a subgroup of patients with brain radiation therapy, EEM-like lesions develop particularly in the radiation field. Most recently, the acronym EMPACT (Erythema Multiforme associated with Phenytoin And Cranial radiation Therapy) was proposed to specifically describe this syndrome. PATIENT/METHOD: Here, we report on EMPACT syndrome in a 46-year-old woman. Therapeutic measures included seizure prophylaxis with phenytoin and total brain radiation therapy of brain metastases from bronchial carcinoma. Three weeks after introduction of phenytoin, the patient presented with EEM-like skin lesions restricted to the original radiation field and facial mucocutaneous involvement. After a few days, the rash spread to the upper part of the body. She was also in poor general condition. RESULTS: The immediate cessation of phenytoin therapy, combined with administration of systemic corticosteroids and high dose immunoglobulins along with intensive local treatment and pain medications, resulted in complete resolution of the skin eruption. Patch testing to phenytoin was positive after 72 hours. CONCLUSION: EMPACT should be classified as an specific entity among the EEM-like drug reactions as it only appears after radiotherapy and seizure prophylaxis with the anticonvulsant phenytoin. We propose including specific type IV-sensitization to phenytoin into the definition of EMPACT. PMID- 16353749 TI - Aggressive undifferentiated squamous cell carcinoma in an immunosuppressed patient after kidney transplantation. AB - A 56-year-old man developed a highly aggressive squamous cell carcinoma (SCC) on the upper back five years ago renal transplantation and subsequent immunosuppression. The tumor was excised using the three-dimensional micrographic technique; after the formation of granulation tissue, the wound was covered with a Meshgraft. His cyclosporine A dose was reduced from 2.5 to 2.0 mg/kg. The patient was counseled on sun avoidance and use of sunscreens, as well as placed on low-dose oral acitretin and imiquimod 5% cream to control actinic keratoses. He is followed every 3 months by a dermatologist and has shown no evidence of recurrence over 2 years. PMID- 16353750 TI - [Gingival Burkitt lymphoma in a hepatitis C-positive renal transplant patient]. AB - Malignant tumors are a frequent complication in the late phase after organ transplantation. Malignant lymphomas are the second most common tumor, exceeded only by cutaneous carcinomas. Compared to the general population, the organ transplant patients have a 30-60 fold increased risk of developing non-Hodgkin's lymphoma. A 55-year-old, hepatitis C-positive man developed an Epstein-Barr virus (EBV)- negative Burkitt lymphoma (BL) first appearing on the gingiva under immunosuppressive therapy nine years after allogenic renal transplantation. He died 37 days after the onset of polychemotherapy. Immediately before his death, meningeal involvement by the BL was diagnosed. BL after organ transplantation is often found at extranodal sites; it involves the central nervous system more frequently than it does in immunocompetent patients. In 70% of BL occurring after organ transplantation, genes or gene products related to EBV can be demonstrated within the tumor cells. The EBV status of the tumor is of important prognostic significance: EBV-positive BL occurring in organ transplant patients usually responds well to reduction or cessation of immunosuppressive therapy; in some cases permanent complete remissions can be achieved even without chemotherapy. In contrast, patients with EBV-negative BL have a very poor prognosis and hardly respond, even to aggressive chemotherapy protocols. PMID- 16353751 TI - [The menstrual cycle and the skin]. PMID- 16353752 TI - [Erosive verrucous skin lesion on the umbilicus of a 83 year old woman]. PMID- 16353753 TI - Porcine skin as an in-vivo model for ageing of human bite marks. AB - Porcine skin has been shown to have similar histological, physiological and immunological properties to human skin and has been suggested as a good analogue for medical and forensic research. This study was undertaken to examine the appearance of bite mark wounds inflicted at known time intervals before and after death. Under general anaesthesia, a series of bite marks were created on a pig's abdomen with a device designed to mechanically produce simulated human bite mark wounds. The pig skin model showed that bite mark characteristics are similar to those found on human skin. This study has provided information on the window of time showing clearly detailed bite marks occurring around the time of death. It also demonstrated that it is possible under certain conditions to determine that a bite mark was made before or after death in a porcine model. Under these experimental conditions, the results suggest that an in-vivo porcine skin model should be considered as a representative model for the study of human bite marks. PMID- 16353754 TI - An historical skull collection and its use in forensic odontology and anthropology. AB - The Institute of Forensic Medicine, Copenhagen, houses a collection of historical skulls of unclear origin, marked with a general geographic or "racial descriptor". Would these historical skulls be of any value for the forensic odontologist and anthropologist concerned with teaching and casework? We tried to clarify this question by recording non-metric dental traits and by performing craniometric analyses. A morphological and morphometric investigation of anatomical/dental traits in 80 adult skulls was performed. For each skull four non-metric dental traits using the ASU-System and three non-metric cranial traits were recorded. Nineteen cranial measures were also taken following the FORDISC programme manual. The non-metrical data were tabulated as frequencies, and the metric data were entered in the FORDISC programme. Observed non-metric trait frequencies were compared with published data. The FORDISC programme computed a discriminatory analysis for each skull and thereby assigned the skull to the most probable ethnic category. The results for the non-metric traits showed that the traits generally followed the expected frequencies in 80% of the cases. The FORDISC programme correctly assigned ethnicity based on skull measurements in overall 70% of the cases. It was found that this historical collection does show expected dental non-metric and craniometric traits and the collection may be of value in forensic casework in terms of comparison and for teaching purposes. PMID- 16353755 TI - Forensic facial approximation: an overview of current methods used at the Victorian Institute of Forensic Medicine/Victoria Police Criminal Identification Squad. AB - Forensic facial approximation involves building a likeness of the head and face on the skull of an unidentified individual, with the aim that public broadcast of the likeness will trigger recognition in those who knew the person in life. This paper presents an overview of the collaborative practice between Ronn Taylor (Forensic Sculptor to the Victorian Institute of Forensic Medicine) and Detective Sergeant Adrian Paterson (Victoria Police Criminal Identification Squad). This collaboration involves clay modelling to determine an approximation of the person's head shape and feature location, with surface texture and more speculative elements being rendered digitally onto an image of the model. The advantages of this approach are that through clay modelling anatomical contouring is present, digital enhancement resolves some of the problems of visual perception of a representation, such as edge and shape determination, and the approximation can be easily modified as and when new information is received. PMID- 16353756 TI - Unusual fatal dog attack in Dunedin, New Zealand. AB - A case of a fatal dog attack on a middle aged woman is presented. The offending dog was her own Bull-mastiff, which had previously shown signs of aggression towards her. Most of the injuries were found on the victim's face, neck and skull. A noteworthy feature of this attack was that the victim was known to suffer from Huntington disease. It is postulated that the involuntary movements, progressive dementia and increased moodiness characteristic of the disease may have had a significant role in triggering the attack. PMID- 16353757 TI - Dento-maxillo-facial radiology as an aid to human identification. AB - Analysis of some anatomical structures of the face using radiographs is fundamental for human identification. The position of the postmortem skull relative to the radiographic machine and the film, as well as the exposure time, are the greatest problems found by the forensic dentist. In view of this fact, some recognised radiographic techniques that are used in vivo must be adapted. This paper shows that simple devices can make the process easier and that variation of kVp or exposure time in different situations may increase the quality of the radiographs. PMID- 16353758 TI - Provide Medicaid to evacuees. PMID- 16353759 TI - If nonprofit doesn't mean "no profit," how much is enough in health care? PMID- 16353760 TI - The effect of SCHIP expansions on health insurance decisions by employers. AB - This study uses repeated cross-sectional data from the Medical Expenditure Panel Survey-Insurance Component (MEPS-IC), a large nationally representative survey of establishments, to investigate the effect of the State Children's Health Insurance Program (SCHIP) on health insurance decisions by employers. The data span the years 1997 to 2001, the period when states were implementing SCHIP. We exploit cross-state variation in the timing of SCHIP implementation and the extent to which the program increased eligibility for public insurance. We find evidence suggesting that employers whose workers were likely to have been affected by these expansions reacted by raising employee contributions for family coverage options, and that take-up of any coverage, generally, and family coverage, specifically, dropped in these establishments. We find no evidence that employers stopped offering single or family coverage outright. PMID- 16353761 TI - The impact of SCHIP on insurance coverage of children. AB - In this paper we use the Medical Expenditure Panel Survey between 1996 and 2002 to investigate the impact of the State Children's Health Insurance Program (SCHIP) on insurance coverage for children. We explore a range of alternative estimation strategies, including instrumental variables and difference-in-trends models. We find that SCHIP had a significant impact in decreasing uninsurance and increasing public insurance for both children targeted by SCHIP and those eligible for Medicaid. With respect to changes in private coverage our results are less conclusive: some specifications resulted in no significant effect of SCHIP on private insurance coverage, while others showed significant decreases in private insurance. Associated estimates of SCHIP crowd-out had wide confidence intervals and were sensitive to estimation strategy. PMID- 16353762 TI - Health plan switching among members of the Federal Employees Health Benefits Program. AB - This paper examines factors associated with switching health plans in the Federal Employees Health Benefits Program. Switching plans is not uncommon, with 12% of members switching plans annually. Individuals switch out of plans with premium increases and benefit decreases relative to other plans in the market. Switching is negatively associated with age due to increasing switching costs associated with age rather than decreasing premium sensitivity. Individuals in preferred provider organizations are less likely to switch, but are more responsive to premium increases than those in the managed care sector. Those who do switch plans are likely to switch to a different plan in the same sector. PMID- 16353763 TI - The value of experience: differences in knowledge among Medicare beneficiaries. AB - The Medicare Advantage program gives Medicare beneficiaries the opportunity to choose from an array of insurance options instead of receiving prescribed benefits. In 2006, beneficiaries who want prescription drug benefits will need to enroll in a Medicare managed care plan or a private prescription drug plan. To examine awareness and use of Medicare information programs, and the extent to which these programs are associated with beneficiary knowledge about Medicare and managed care, we conducted a national survey of Medicare beneficiaries six to 12 months after the nationwide mailing of the Medicare & You 2000 handbook. Beneficiary information-gathering behavior and experience with Medicare managed care were more highly associated with knowledge about Medicare managed care than formal education, age, income, or membership in a managed care plan before enrolling in Medicare. Practical life experience appears to outweigh traditional factors in beneficiary knowledge of Medicare and managed care. PMID- 16353764 TI - A copayment increase for prescription drugs: the long-term and short-term effects on use and expenditures. AB - This study estimates the effects of an increase in an outpatient prescription drug copayment using a natural experiment based upon a large firm that implemented such an increase. The findings suggest that the primary effect of a copayment increase is attenuation of the trend in prescription drug utilization. We also find an initial reduction in expenditures, with the effects on spending diminishing. Employees with an existing chronic illness and those without a chronic illness show a similar, inelastic response to a copayment increase; employees with a newly diagnosed chronic illness have a more inelastic response. PMID- 16353765 TI - [100th anniversary of Dr D. Antonio Puigvert Gorro's birth]. PMID- 16353766 TI - [Blood, hemorrhage, hematuria, urology]. PMID- 16353767 TI - [High risk disseminated prostate cancer (consensus paper and recommendations)]. PMID- 16353768 TI - [Influence of climate, according to Koppen classification and Lang's index, on gross rate mortality associated with prostate cancer in the geographical area of Spain]. AB - OBJECTIVE: To evaluate if the climatic factors of temperature and rainfall influence mortality associated with prostate cancer for a period of five years. MATERIAL AND METHODS: The mortality trends associated with prostate cancer will be evaluated in the period ranging from 1st january 1998 to 31st december 2002, in the geographical area of Spain. The demographic and mortality data were obtained from the National Institute of Statistics and the climatologically data on temperature and rainfall were obtained from the National Meteorology Institute applying the values registered in 2002. Based on the meteorological data, the different provinces are classified in accordance with Koppen climate classification and climatic index of Lang that defines them from the mean annual and monthly temperature and rainfall data. Mortality rates will be expressed by 100.000 men year, applying for their study the Kruskall-Wallis test for "n" independent samples through the SPSS v12.0 Windows software. RESULTS: Three main climatic areas are obtained, one area with type B climate, which accounts for 13% of the total population, a Csa climate, and a Csb, accounting for 67 and 20% respectively. The mortality found is higher in the Csb type for prostate cancer (p= 0.007), as compared to the other two climatic areas. CONCLUSION: Mortality associated with prostate cancer is significantly higher in the regions with a lower sun exposure. However, randomized prospective studies confirming these findings are warranted. PMID- 16353769 TI - [Dose escalation with three dimensional conformal radiotherapy for prostate cancer: more is better?]. AB - PURPOSE: The present study was undertaken to determine the effect of radiation dose on biochemical control and morbidity in prostate cancer patients. MATERIALS AND METHODS: Between 1995 and 2003, 360 patients with T1-T3b prostate cancer were treated in a sequential radiation dose escalation trial from 66.0 to 82.6 Gy. These patients were prospectively assigned to 1 of 3 prognostic groups according to risk factors: a) low risk patients were treated with 3DCRT alone; b) intermediate risk patients were allocated to receive neoadjuvant AD (NAD) 4-6 months prior and during 3DCRT; and c) high-risk received NAD and adjuvant AD (AAD) 2 years after 3DCRT. RTOG/EORTC toxicity score was used to analyze late complications RESULTS: Median follow-up was 48 months (12-138). The actuarial biochemical disease free survival (bDFS) at 4 years for low risk, intermediate risk and high risk patients was 88%, 68% and 79% respectively. Stratified and multivariate analysis showed that higher radiation dose (>76 Gy) (p=0.0053) and the use of AAD for high risk patients (p=0.0046) correlated significantly with an improvement of bDFS for all patients. The incidence of late grade 2 rectal and urinary bleeding were 7% and 11% respectively. CONCLUSION: The present study confirms an independent benefit of high-dose (> 76 Gy) radiation therapy and long term AAD in high-risk prostate cancerpatients. PMID- 16353770 TI - [Is there a place for randomized biopsies in the follow up of superficial transitional cell carcinoma of the urinary bladder?]. AB - INTRODUCTION: Transitional cell carcinoma of the bladder represents a disease of entire urothelial tract. The follow up is very important to detect any lesion that might represent a progression or a local recurrence. Some authors recommend randomized biopsies as a routine workup, others recommend cystoscopies and urinary cytology as the main part of superficial bladder cancer follow up. PATIENTS AND METHODS: Forty nine patients with superficial bladder cancer were followed up during a ten-year period. Randomized biopsies and urinary cytology were harvested according to the international cancer protocol on bladder cancer. RESULTS: 15 (1%) out of 1.489 randomized biopsies found to be positive to transitional cell carcinoma. Four out (10.5%) of 35 biopsies targeted to suspicious areas were positive to transitional cell carcinoma. 50 (17.4%) out of 288 cystoscopies with urinary cytology found to be positive to transitional cell carcinoma. Sensitivity and Specificity of biopsies (including randomized and targeted) were 31% and 85.2% respectively. Sensitivity and specificity of cystoscopies with urinary cytology were 48% and 86.5% respectively. CONCLUSION: Randomized biopsies did not show to detect more local recurrence or progression when compared to the urinary cytology. Cystoscopies with urinary cytology have good sensitivity and specificity for detection of tumor recurrence during follow up of transitional cell carcinoma. PMID- 16353771 TI - [Cell cycle and apoptosis mechanisms implicated in intravesical chemotherapy resistances in superficial bladder cancer]. AB - It is well documented the effectiveness of intravesical chemotherapy following transurethral resection to prevent recurrences of superficial bladder cancer. But it is also known that efficacy may be limited by tumour cell resistance to one or several of the drugs available for instillation. In addition to the genetically determined unicellular mechanisms classically described in the literature such as glycoprotein P-170 expression (mdr-1), overexpression of Bcl-2 or glutation S transferase activity, it has been recently shown that multicellular mechanisms may also be involved in drug resistance. Multicellular resistance can only be demonstrated in three-dimensional cultures and fails to be shown in monolayers or cell suspensions. This is explained by the fact that cell-to-cell and cell-to stroma adhesion limits drug penetration and by the variable susceptibility to cytotoxicity determined by oxygen and tissue proliferation gradients. A better understanding of the molecular mechanisms involved in drug resistance is necessary to increase intravesical chemotherapy effectiveness. Current research includes improving drug penetration, searching resistance reversing agents and developing in vitro chemosensitivity tests to identify drug resistance. PMID- 16353772 TI - [Laparoscopic assisted cryoablation of small renal masses]. AB - AIMS: To review the current status of cryoablation of small renal masses and to preliminary report our experience at the AMC. MATERIAL AND METHODS: A bibliographic search was conducted (PubMed/Medline/Embase) and the most important series were analyzed. Our series includes 13 patients with a solitary small renal mass treated by Laparoscopic assisted Cryoblation with fine cryoprobes (1,5 cm diameter). Postoperative follow-up was done by means of CT and/or MRI every three months during the first year and every 6 months during the second year. RESULTS: There are no randomized trials comparing Cryoblation of renal masses (<4 cm diameter) with either radical surgery or partial nephrectomy but only case series. Complication rate is low as it is the recurrence or persistence rate in most of the series but one referring to radiologically guided Cryoablation (8%). The maximal diameter of the masses treated in our series were 3,2 cm. Tumors were approached retro or transperitoneally depending on their localization in the kidney. Average surgical time were 208 minutes (108-379) and average time of exposition to temperatures lower than -20 degrees C in the tumor periphery was 10 minutes. AT a mean follow-up of 8 months no tumoral recurrence were objectivated. CONCLUSIONS: Cryoablation of small renal masses may be an acceptable alternative of treatment although mean follow-up is still short in all the series. PMID- 16353773 TI - [Spontaneous urinary bladder perforation after pediatric cystoplasty]. AB - OBJECTIVE: The spontaneous cystoplasty perforation is a serious and potentially fatal problem if a delay in diagnosis and treatment occurs. We pretend: 1) to look for prevention analyzing the risk factors, 2) to identify the main data of diagnostic suspicion and 3) to evaluate the result of the treatments done. MATERIAL AND METHODS: Out of 30 children with cystoplasty 5 of them have presented 8 perforations (16,6%). Several influential factors, the symptoms, the treatments and the evolution are reviewed. RESULTS: The average time between cystoplasty and the perforation was 8,2 years. A urethral resistance that allows continence, and an insufficient intermittent catheterization, have been the main risk factors. In the 8 episodes there were abdominal pain and distension. The ultrasonography showed intraperitoneal extravasation in 5 episodes, multiple peritoneal cysts in one, and suggestive image of appendicular plastron in another one. The cystography showed intraperitoneal extravasation only in 3 cases. The initial management was conservative in the 7 episodes diagnosed before surgery, and 3 had a good evolution (42,8%); the other 4 needed surgery with good evolution in all cases. Two of 5 patients (40%) presented 3 relapses in an average time of 5 years. The survival is 100%. CONCLUSIONS: 16,6% of patients with cystoplasty of this series had one or more episodes of spontaneous bladder perforation. The more significant risk factors are a high urethral resistance and an inadequate intermittent catheterization. The patients with cystoplasty, and their families, must know this complication, their risk factors and symptoms to prevent it, or to facilitate an early diagnosis. PMID- 16353774 TI - [Correlation between urogenital prolapse and detrusor overactivity]. AB - PURPOSE: To evaluate the urodynamic characteristics of a series of women with stress urinary incontinence presenting various rates of vaginal prolapse. METHODS: 92 female patients were included in this study, mean age was 48 years (range 29-75). All patients underwent urogynecologic physical examination (vaginal prolapse was graded) and complete urodynamic study in order to detect the presence of detrusor overactivity. Urodynamic terminology and measurements comply with the ICS (International Continence Society) standards. Statistical significance was established below 0.05. RESULTS: 65 women (70.6%) presented anterior vaginal prolapse and 31 (33.6%) posterior vaginal prolapse. Involuntary contractions of the detrusor muscle appeared in 13 patients (20%) who had an anterior vaginal prolapse and 6 women (19.3%) who had a posterior vaginal prolapse. The existence of involuntary contractions was not associated with the diagnosis of vaginal prolapse. CONCLUSION: Our study did not show any correlation between existence of vaginal prolapse and detrusor overactivity. PMID- 16353775 TI - [Psychological features and Doppler-Duplex in erectile disfunction]. AB - OBJECTIVES: To establish a relationship between Doppler-Duplex colour ultrasound after prostaglandin intracorporeal injec tion and psychological features in patients suffering from erectile dysfunction. METHODS: Forty two patients with erectile dysfunction were prospectively evaluated with Doppler-Duplex colour ultra sonography after intracavernosal injection of 20 micrograms of E1 prostaglandin. Dynamic vascular pattern were analyzed an penile tumescence were graded in poor, moderate or good (I, II, III). All of them completed the International Index of Erectile Dysfunction, International Exam of Personality Traits (IPDE) and the Symptom Checklist (SCL-90). RESULTS: 29 patients (69.05%) showed a normal ultrasonography response (Peak Systolic Velocity >30 cm/s; Telediastolic velocity negative or less than 5 cm/s and penile tumescence grade III) and were classified as good responders with probably psy chogenic erectile dysfunction. Patients who did not show these values were classified in the group of erectile dysfunction of vas cular origin. We found significative differences between the two groups in sex desire, tumescence and paranoid personality. Th more frequent personality features were paranoid and squizoid (excentric) and ananchastic and anxious (fearful ones). Somatization and obsessive-compulsive symptoms have been outlined over the rest. We have found that paranoid personality is sixteen times more frequent in patients with normal ultrasonography. CONCLUSIONS: Psychological features and dysfunctional personality traits accompany psychogenic and organic erectile dys function, thus it is thought that mixed aetiology coexist in this patients. Doppler-Dupplex Colour ultrasound is an usefu method to exclude organic vascular factors. On de basis of our article, is more common to find psychological discomfort an dysfunctional personality traits in patients with normal ultrasound, which may help in their diagnosis and treatment. PMID- 16353776 TI - [Erectile dysfunction: the role of laboratory in the diagnostic and pronostic evaluation]. AB - OBJECTIVE: To review and to update the different laboratory tests recommended for etiologic diagnostic of erectile dysfunction and to evaluate the effect these tests could have on the pronostic and therapeutic strategy of this pathology. MATERIAL AND METHODS: We review the last articles related with etiopathogenics and pathophysiologics mechanisms of erectile dysfunction, including our studies on endothelial dysfunction and erectile dysfunction. RESULTS: The depth and extension of the laboratory protocol in erectile dysfunction is not necessaryly the same in all situations. The age, coincidence of comorbilities, set a different limit between patients demanding complementaries investigations that go beyond the basic request. CONCLUSIONS: The etiopathogenic laboratory work up in erectile dysfunction is currently changing incorporating news tests. The traditional search of commorbilities like diabetes, hepatic dysfunction, hypogonadism, hyperglucemia is getting broad with recents analitics evaluations related with potential markers of endothelial disease. PMID- 16353777 TI - [Primary small cell carcinoma of the bladder treated with partial cystectomy. Case report]. AB - Primary small cell carcinomas of the bladder are extremely rare tumors with an aggressive behavior. To date, no standard therapy has been proposed. There are some brief reports about the role of conservative surgery in patients not suitable for radical surgery. We report a case of a primary bladder small cell carcinoma treated with partial cystectomy. PMID- 16353778 TI - [Primary non-Hodgkin large B-cell lymphoma of bladder. Report of a case]. AB - INTRODUCTION: primary genitourinary lymphomas are uncommon. Among them, bladder lymphomas are extremely unusual tumors, with clinico-radiological features similar to urothelial carcinomas of bladder. Histopathological, immunohistochemical and molecular studies are compulsory for the diagnosis. We report a case of this tumor. CLINICAL CASE: An 80-year-old woman was admitted to our hospital with hematuria. Abdominal ultrasound and cystoscopy revealed an infiltrating bladder tumor involving the right lateral wall. After transuretral biopsy, a diagnosis of non-Hodgkin large B-cell lymphoma was made. Neither clinical symptoms nor radiological findings showed disseminated disease, indicating that the tumor was localized in the bladder. After chemotherapy, the patient is disease-free after 9 months follow-up. COMMENT: if a bladder tumor with uncommon histopathological features is found, lymphoma should be excluded, because chemotherapy avoids cystectomy. PMID- 16353779 TI - [Testicular epidermoid cyst: uncommon lesion of difficult preoperative diagnosis]. AB - Intratesticular epidermoid cysts are rare tumours that constitute one percent of all testicular masses. They are bening lesions that make differential diagnosis from malignant testicular tumours difficult. The absence of serum markers elevation and ultrasound imaging could support these lesions being bening epidermoid cysts, and in that case, conservative surgery is adequate. We present the case of a 22 years old patient who complains of a left testicular mass. In this case ultrasound diagnosis was non-specific and a left radical inguinal orchiectomy was performed. PMID- 16353780 TI - [Urachal cyst presenting as acute urinary retention]. AB - About 2% of adults have an urachal cyst. The diagnosis is usually made due to its clinical complications. We report an inusual case of acute urinary retention due to an urachal cyst, with hipogastric pain and anuria as initial syntoms. PMID- 16353781 TI - [The impact factor. Actas Urologicas Espanolas]. PMID- 16353782 TI - [Crossed renal ectopy with fusion]. PMID- 16353783 TI - [Esthesioneuroblastoma treatment]. AB - INTRODUCTION: Esthesioneuroblastoma is an uncommon malignancy of the olfactory neuroepithelium. The best treatment has yet to be defined. The purpose of this study is to analyze the tumors's behaviour to choose the ideal treatment, the therapeutic strategy and the patterns of failure. MATERIALS AND METHODS: We carry out a revision of the series published between 1994 to 2004. In these series, we found 39 papers with 713 patients. DISCUSSION: CONCLUSIONS: In this review the 5 years survival rate is 51.2%. Through the analysis of 583 partients found in 34 publications, surgery (alone or combined) is the treatment most used (78%). The commonly management is surgical in combination with radiotherapy (47%). PMID- 16353784 TI - [Cochlear involvement in acute otitis media]. AB - OBJECTIVE: An experimental design with mice has been realized to understand the origin of sensorial hearing loss related to infectious processes in the middle ear. Our aim is to study if the cilliated cells in the Corti organ's are lost after one or more acute otitis media. MATERIAL AND METHODS: For that, thirty Sprague-Dawley mice have been divided in two groups. In the first one, we have induced one acute otitis media, in the second group we have induced two infections of this kind. RESULTS: The statistic study shows that the loss of the cilliated cells is greater in the basal turn of the mice is cochlea, in the first group (alpha=0,020) and also in the second (alpha=0,038). We've noticed in the first group the near absence of injury in the middle turn, but in all samples of the second group we found that injury (alpha=0,002). CONCLUSIONS: In acute otitis media, the loss of ciIliated cells are mainly in the basal turn of the mice is cochlea, and affects the middle turn when the infection is repeated. PMID- 16353785 TI - [Stapedectomy: our results]. AB - OBJECTIVE: The aim of this study is to value the hearing results obtained in our department following stapedectomy and the reasons of primary failures. MATERIAL AND METHOD: We present a retrospective study of 95 stapedectomies in 82 patients, performed from January 1997 to September 2002 at Hospital Xeral-Cies of Vigo (Pontevedra). RESULTS: Medium presurgical audiometric threshold was 65.71 and postsurgical 26.68. Good results were obtained in 84.21% of cases (air-bone gap closure difference under 20 dB), 10.52% had not change in air threshold and the rest deteriorated. CONCLUSIONS: We compare our results with other authors and we consider that stapedectomy is a safe surgical technique, but it has important complications and they have to be bear in mind to prevent them. PMID- 16353786 TI - [Computational fluid dynamics simulations of the airflow in the human nasal cavity]. AB - OBJECTIVE: This study represents an attempt to simulate the complex three dimensional airflow pattern in the human nasal passageways. MATERIALS AND METHODS: In the present study the computational Fluid Dynamics (CFD) have been used. The CFD solved numerically the flow governing equations. The CFD demonstrated to be a very efficient tool for researching on nasal flow. RESULTS: By means of CT scan, an anatomic accurate three-dimensional representation of the human nasal cavity was obtained. Based on this model the influence of the inspiration and at the expiration at the flow pattern has been reviewed. The flow pattern is visualized by the velocity, pressure and turbulence intensity fields. CONCLUSIONS: The flow patterns show the channel effect of turbinates and the influence of the breathing phase. PMID- 16353787 TI - [Gastroesophageal reflux and obstructive sleep apnea syndrome]. AB - INTRODUCTION: Patients with mild or moderate Sleep Apnea Syndrome (SAS) need wider therapeutic scope options according to their disease severity. AIM: To consider including proton pump inhibitors (PPI) to the therapeutical alternatives of these patients. MATERIAL AND METHODS: A prospective study was designed, among patients with SAS. Nocturnal polysomnography and double channel pHmetry were performed simultaneously. RESULTS: From the 18 patients included in this preliminary phase, in three (16.7%) nocturnal proximal ph monitoring was positive. These 3 patients were treated with PPI during at least 3 months with a very satisfactory outcome in two of them. CONCLUSIONS: Treatment with PPI may be a useful therapeutical alternative in patients with mild to moderate SAS. PMID- 16353788 TI - [Functional and oncologic results of the surgical salvage in the management of recurrent nasopharyngeal carcinomas]. AB - PURPOSE: To evaluate the indications and results of the surgical salvage in the treatment of recurrent nasopharyngeal carcinoma at the primary site. MATERIAL AND METHOD: Twelve patients with biopsy-proven recurrent nasopharyngeal cancer were managed with the subtemporal-preauricular and facial translocation approaches. One patient was staged as having recurrent T stage (rT1) 1 disease; 3 patients, rT2; 2 patients, rT3; and 6 patients rT4. RESULTS: With a mean follow-up of 3 years, the 3-year actuarial survival rate was 42%. Survival for recurrent T stage was as follows: rT1, 100%; rT2, 67%; rT3, 50%; rT4, 17%. Survival rates for patiens with rT3 and rT4 disease without intracranial disease are 40%. CONCLUSIONS: Facial translocation and subtemporal-preauricular approaches are versatile surgical techniques for radical resection of nasopharyngeal recurrences with an acceptable morbidity. The long term survival results are encouraging for rT1-rT3 tumors and also for rT4 without intracranial extension of the disease. PMID- 16353789 TI - [Near-total laryngectomy. Experience, complications and acoustic analysis]. AB - OBJECTIVE: To investigate the oncological efficiency, voice and complications of Pearson's near-total laryngectomy for advanced laryngeal cancers (T3, T4). MATERIAL AND METHODS: A retrospective review of 23 cases of near-total laryngectomy (carried out in our Hospital in the last nine years) was undertaken. RESULTS: None of the cases developed laryngeal mucosal recurrences. The overall three and five years disease free survival were 91,3% and 86,9%. Mean phonation time was 51 days. A statistically significant results were observed in our voice analysis (words/minute, taking as a reference healthy people (Medivoz). The pharyngeal fistula was the most frequent complication (60,86%). CONCLUSIONS: The local control of cancer was similar to that expected with total laryngectomy. Near-total laryngectomy is a valid alternative for extended laryngeal and neighbouring cancers with an acceptable morbidity and success rate for voice preservation. PMID- 16353790 TI - [The relationship between human papillomavirus and cyclin D1 gene amplification in squamous cell carcinomas of the head and neck]. AB - INTRODUCTION: infection by oncogenic subtypes of human papillomavirus (HPV) and cyclin D1 gene (CCND1) amplification are frequent events in head and neck squamous cell carcinomas. The objective of this paper is to establish the relationship between the presence of human papillomavirus (HPV) gene sequences and the development of CCND1 gene amplification in these tumours. MATERIALS AND METHODS: 59 squamous cell carcinomas of the head and neck were studied for HPV types 6b and 16 and CCND1 gene amplification by polymerase chain reaction. RESULTS: HPV DNA was detected in 14 tumors (24%). Ten of them were positive for the HPV type 6b and 4 for the HPV type 16. CCND1 gene amplification was found in 15 cases (25%). Although we have found a higher frequency of CCND1 amplification in the HPV-positive cases (36%, versus 22% in the HPV-negative cases), these differences were not statistically significant (P= 0,32). CONCLUSIONS: The presence of HPV gene sequences does not seem to be related to a significative higher incidence of CCND1 gene amplification in the squamous cell carcinomas of the head and neck. PMID- 16353791 TI - [Intrapretosal schwannomas of facial nerve]. AB - The aim of this study was to determine the factors that are important to the diagnosis and treatment of facial neuroma. The most common tumors of the peripheral nerves are the neurinomas; about 50% of them are found in the head and neck region. Among them, the most frequent is the VIII nerve tumor, while the facial neuroma is extremely rare. Neuromas can be intracraneal, intratemporal and extratemporal. The last one has a higher frequency. We present our experience in the intrapetrosal facial neuromas; usually showing different presentation symptoms wich is a characteristic of them. Retrospective study that was carried out at a private tertiary referral center. 3 patients were referred from other centers for otological surgery. One of them had symptoms of chronic otitis media, the second one was diagnosed of a glomus tumor and the last one showed a facial palsy. All the patients were operated on and then the facial nerve was repaired. Two patients had an end to end anastomosis, and in the other one we managed to keep the integrity of the facial nerve. The Hous -Brackmann facial grading scale was used. Two patients had grade II and the other grade III in this scale in the long-term follow up. PMID- 16353792 TI - [Metastasis in the palatine tonsil as the first sign of a gastric adenocarcinoma]. AB - Gastric adenocarcinomas rarely give rise to metastases in the palatine tonsils; twelve such cases have been published. In two of them the tonsillar tumour was the first sign of the disease. We present a patient with a tumour of the palatine tonsil and cervical lymph node involvement who was diagnosed as having an adenocarcinoma of "signet ring" cells in the histopathology and cytology studies. Finding such cells directed us to the stomach in our search for the primary. PMID- 16353793 TI - Phase demodulation method from a single fringe pattern based on correlation with a polynomial form. AB - The method presented extracts the demodulated phase from only one fringe pattern. Locally, this method approaches the fringe pattern morphology with the help of a mathematical model. The degree of similarity between the mathematical model and the real fringe is estimated by minimizing a correlation function. To use an optimization process, we have chosen a polynomial form such as a mathematical model. However, the use of a polynomial form induces an identification procedure with the purpose of retrieving the demodulated phase. This method, polynomial modulated phase correlation, is tested on several examples. Its performance, in terms of speed and precision, is presented on very noised fringe patterns. PMID- 16353794 TI - Characterization of gradient-index lens-fiber spacing toward applications in two photon fluorescence endoscopy. AB - We report on the experimental investigation into the characterization of two photon fluorescence microscopy based on the separation distance of a single-mode optical fiber coupler and a gradient-index (GRIN) rod lens. The collected two photon fluorescence signal exhibits a maximum intensity at a defined separation distance (gap length) where the increasing effective excitation numerical aperture is balanced by the decreasing confocal emission collection. A maximum signal is found at gap lengths of approximately 2, 1.25, and 1.75 mm for GRIN lenses with pitches of 0.23, 0.25, and 0.29 wavelength at 830 nm. The maximum two photon fluorescence signal collected corresponds to a threefold reduction of axial resolution (38.5 microm at 1.25 mm), compared with the maximum resolution (11.6 microm at 5.5 mm), as shown by the three-dimensional imaging of 10 microm beads. These results demonstrate an intrinsic trade-off between signal collection and axial resolution. PMID- 16353795 TI - Assessment of refractive index gradients by standard rainbow thermometry. AB - Up to now the application of rainbow thermometry has been limited to particle systems possessing a uniform refractive index. This is mostly due to the absence of an appropriate data inversion algorithm that takes into account the presence of a refractive index gradient. In this paper, for the first time to our knowledge, exploiting a generalization of the Airy theory, a data inversion algorithm for a single droplet, presenting a parabolic refractive index gradient, is proposed. This data inversion algorithm is used to compute the diameter and the refractive index at the core and at the surface of a simulated burning droplet. The results are compared to the analytical solutions showing a satisfactory agreement. PMID- 16353796 TI - Optical vector network analyzer for single-scan measurements of loss, group delay, and polarization mode dispersion. AB - We present a method for measuring the complete linear response, including amplitude, phase, and polarization, of a fiber-optic component or assembly that requires only a single scan of a tunable laser source. The method employs polarization-diverse swept-wavelength interferometry to measure the matrix transfer function of a device under test. We outline the theory of operation to establish how the transfer function is obtained. We demonstrate the enhanced accuracy, precision, and dynamic range of the technique through measurements of several components. PMID- 16353797 TI - In-line interferometer for direction-sensitive displacement measurements by optical feedback detection. AB - We demonstrate a compact in-line interferometer for direction-sensitive displacement measurement by optical feedback detection with a semiconductor laser (SL) light source. Two reflected beams from a semitransparent reference mirror and a reflecting test object interfere in the SL medium, causing a variation in its output power. The reference mirror is located between the SL output facet and the test object. The performance of the interferometer is investigated numerically and experimentally to determine its optimal operating conditions. We have verified the operating conditions where the behavior of the SL output power profile could indicate accurately the displacement magnitude and direction of the moving test object. The profile behavior is robust against variations in optical feedback and scale of the interferometer configuration. PMID- 16353798 TI - Plasma interferometry and how the bound-electron contribution can bend fringes in unexpected ways. AB - Utilizing a new average atom code, we calculate the index of refraction in C, Al, Ti, and Pd plasmas and show many conditions over which the bound-electron contribution dominates the free electrons as we explore photon energies from the optical to 100 eV (12 nm) soft x rays. For decades measurement of the electron density in plasmas by interferometers has relied on the approximation that the index of refraction in a plasma is due solely to the free electrons and therefore is less than 1. Recent measurements of Al plasmas using x-ray laser interferometers observed fringes bending in the opposite direction than expected due to the bound-electron contribution causing the index of refraction to be larger than 1. During the next decade x-ray free-electron lasers and other sources will be available to probe a wider variety of plasmas at higher densities and shorter wavelengths, so understanding the index of refraction in plasmas is essential. PMID- 16353799 TI - Rapid subsurface detection of nanoscale defects in live microprocessors by functional infrared emission spectral microscopy. AB - We demonstrate the rapid and nondestructive detection of subsurface nanometer size defects in 90 nm technology live microprocessors with a new technique called functional infrared emission spectral microscopy. Broken, leaky, and good transistors with similar photoemission images are identified from each other by their different emission spectra that are calculated as linear combinations of weighted basis spectra. The basis spectra are derived from a spectral library by principal component analysis. Leaky transistors do not exhibit apparent morphological damage and are undetectable by optical or scanning probe microscopy alone. The emission signals from two or more transistors combined incoherently, and defect detection is primarily limited by the signal-to-noise ratio of the detected spectrum and not by the separation distance of neighboring transistors. PMID- 16353800 TI - Finite conjugate spherical aberration compensation in high numerical-aperture optical disc readout. AB - Spherical aberration arising from deviations of the thickness of an optical disc substrate from a nominal value can be compensated to a great extent by illuminating the scanning objective lens with a slightly convergent or divergent beam. The optimum conjugate change and the amount and type of residual aberration are calculated analytically for an objective lens that satisfies Abbe's sine condition. The aberration sensitivity is decreased by a factor of 25 for numerical aperture values of approximately 0.85, and the residual aberrations consist mainly of the first higher-order Zernike spherical aberration term A60. The Wasserman-Wolf-Vaskas method is used to design biaspheric objective lenses that satisfy a ray condition that interpolates between the Abbe and the Herschel conditions. Requirements for coma by field use allow for only small deviations from the Abbe condition, making the analytical theory a good approximation for any objective lens used in practice. PMID- 16353801 TI - Subwavelength surface-relief gratings for stellar coronagraphy. AB - We present a new design of a phase mask coronagraph implemented with subwavelength diffractive optical elements consisting of optimized surface-relief gratings. Phase mask coronagraphy is a recent technique that seeks to accommodate both high dynamic and high angular resolution imaging of faint sources around bright astrophysical objects such as exoplanets orbiting their host stars. The original design we propose is a new, integrated, and flexible solution to the pi phase-shift chromaticity of the phase mask coronagraphs. It will allow broadband observations, i.e., shorter integration times and object characterizations, by means of spectroscopic analysis. The feasibility of the component manufacturing is also considered through a tolerance study. PMID- 16353802 TI - Active optics and modified-Rumsey wide-field telescopes: MINITRUST demonstrators with vase- and tulip-form mirrors. AB - Wide-field astronomy requires the development of larger aperture telescopes. The optical properties of a three-mirror modified-Rumsey design provide significant advantages when compared to other telescope designs: (i) at any wavelength, the design has a flat field and is anastigmatic; (ii) the system is extremely compact, i.e., it is almost four times shorter than a Schmidt. Compared to the equally compact flat-field Ritchey-Chretien with a doublet-lens corrector, as developed for the Sloan digital sky survey-and which requires the polishing of six optical surfaces-the proposed modified-Rumsey design requires only a two surface polishing and provides a better imaging quality. All the mirrors are spheroids of the hyperboloid type. Starting from the classical Rumsey design, it is shown that the use of all eight available free parameters allows the simultaneous aspherization of the primary and tertiary mirrors by active optics methods from a single deformable substrate. The continuity conditions between the primary and the tertiary hyperbolizations are achieved by an intermediate narrow ring of constant thickness that is not optically used. After the polishing of a double vase form in a spherical shape, the primary-tertiary hyperbolizations are achieved by in situ stressing. The tulip-form secondary is hyperbolized by stress polishing. Other active optics alternatives are possible for a space telescope. The modified-Rumsey design is of interest for developing large space- and ground based survey telescopes in UV, visible, or IR ranges, such as currently demonstrated with the construction of identical telescopes MINITRUST-1 and -2, f/5-2 degrees field of view. Double-pass optical tests show diffraction-limited images. PMID- 16353803 TI - Characterization of AlF3 thin films at 193 nm by thermal evaporation. AB - Aluminum fluoride (AlF3) was deposited by a resistive heating boat. To obtain a low optical loss and high laser-induced damage threshold (LIDT) at 193 nm, the films were investigated under different substrate temperatures, deposition rates, and annealing after coating. The optical property (the transmittance, refractive index, extinction coefficient, and optical loss) at 193 nm, microstructure (the cross-sectional morphology, surface roughness, and crystalline structure), mechanical property (stress), and LIDT of AlF3 thin films have been studied. AlF3 thin films deposited at a high substrate temperature and low deposition rate showed a lower optical loss. The highest LIDT occurred at the substrate temperature of 150 degrees C. The LIDT of the films prepared at a deposition rate of 2 A/s was higher than that at other deposition rates. The annealing process did not influence the optical properties too much, but it did increase the LIDT and stress. PMID- 16353804 TI - Complex-argument Laguerre-Gauss beams: transport of mean-squared beam width. AB - For the azimuthally varying complex-argument Laguerre-Gauss beams, the Fourier integral representation is used to obtain the time-averaged power and the transport equation for the mean-squared beam width. From the coefficients in the transport equation, two propagation parameters are derived and compared with the previous treatments. PMID- 16353805 TI - Polarization holographic high-density optical data storage in bacteriorhodopsin film. AB - Optical films containing the genetic variant bacteriorhodopsin BR-D96N were experimentally studied in view of their properties as media for holographic storage. Different polarization recording schemes were tested and compared. The influence of the polarization states of the recording and readout waves on the retrieved diffractive image's intensity and its signal-to-noise ratio were analyzed. The experimental results showed that, compared with the other tested polarization relations during holographic recording, the discrimination between the polarization states of diffracted and scattered light is optimized with orthogonal circular polarization of the recording beams, and thus a high signal to-noise ratio and a high diffraction efficiency are obtained. Using a He-Ne laser (633 nm, 3 mW) for recording and readout, a spatial light modulator as a data input element, and a 2D-CCD sensor for data capture in a Fourier transform holographic setup, a storage density of 2 x 10(8) bits/cm2 was obtained on a 60 x 42 microm2 area in the BR-D96N film. The readout of encoded binary data was possible with a zero-error rate at the tested storage density. PMID- 16353806 TI - Blind multichannel reconstruction of high-resolution images using wavelet fusion. AB - We developed an approach to the blind multichannel reconstruction of high resolution images. This approach is based on breaking the image reconstruction problem into three consecutive steps: a blind multichannel restoration, a wavelet based image fusion, and a maximum entropy image interpolation. The blind restoration step depends on estimating the two-dimensional (2-D) greatest common divisor (GCD) between each observation and a combinational image generated by a weighted averaging process of the available observations. The purpose of generating this combinational image is to get a new image with a higher signal-to noise ratio and a blurring operator that is a coprime with all the blurring operators of the available observations. The 2-D GCD is then estimated between the new image and each observation, and thus the effect of noise on the estimation process is reduced. The multiple outputs of the restoration step are then applied to the image fusion step, which is based on wavelets. The objective of this step is to integrate the data obtained from each observation into a single image, which is then interpolated to give an enhanced resolution image. A maximum entropy algorithm is derived and used in interpolating the resulting image from the fusion step. Results show that the suggested blind image reconstruction approach succeeds in estimating a high-resolution image from noisy blurred observations in the case of relatively coprime unknown blurring operators. The required computation time of the suggested approach is moderate. PMID- 16353807 TI - Finite element formulation for a digital image correlation method. AB - A finite element formulation for a digital image correlation method is presented that will determine directly the complete, two-dimensional displacement field during the image correlation process on digital images. The entire interested image area is discretized into finite elements that are involved in the common image correlation process by use of our algorithms. This image correlation method with finite element formulation has an advantage over subset-based image correlation methods because it satisfies the requirements of displacement continuity and derivative continuity among elements on images. Numerical studies and a real experiment are used to verify the proposed formulation. Results have shown that the image correlation with the finite element formulation is computationally efficient, accurate, and robust. PMID- 16353808 TI - Intensity fluctuations in laser links between the ground and a satellite. AB - Laser-beam-intensity fluctuations between an uplink (ground to satellite) and a downlink (satellite to ground) are described. The dependence of the beam radius on log-intensity variance in the uplink and the dependence of receiving-aperture diameter on the log-intensity variance in the downlink using a known theory were calculated. Statistical analysis of the experimental data on the laser links was performed, and high correlation coefficients of 0.94 between the normalized intensity variances of the uplink and those of the stellar scintillation were obtained. The beam-pointing error in the satelliteborne laser transmitter chiefly caused fluctuations in the downlink. PMID- 16353809 TI - Narrow band fiber-optic phase-shifted Fabry-Perot Bragg grating filters for atmospheric water vapor lidar measurements. AB - A unique ultranarrowband fiber-optic phase-shifted Fabry-Perot Bragg grating filter for atmospheric water vapor lidar measurements was designed, fabricated, and successfully tested. Customized optical fiber Bragg gratings were fabricated so that two transmission filter peaks occurred: one (89% transmission, 8 pm FWHM) near the 946-nm water vapor absorption line and the other peak (80% transmission, 4 pm FWHM) at a region of no absorption. Both transmission peaks were within a 2.66-nm stop band. Demonstration of tension tuning to the 946.0003-nm water vapor line was achieved, and the performance characterization of custom-made optical fiber Bragg grating filters are presented. These measurements are successfully compared to theoretical calculations using a piecewise-matrix form of the coupled mode equations. PMID- 16353810 TI - Reevaluation of the quondam dust trend in the middle atmosphere. AB - Quondam lunar eclipse photometry data offered valuable information on the optical properties of the middle atmosphere, including dust particles. However, in comparison with nonspherical grains, the simple model of spherical particles has a different effect on solar radiation penetrating horizontally through the atmosphere. It is shown that the systems, in which the smallest size fraction of dust particles dominates, reduce irradiation of the Earth's shadow more efficiently if the grains are of irregular shape. In contrast, the populations contaminated by a certain amount of large particles cause an opposite effect. Depending on the actual form of the size distribution function of the irregular grains, the irradiance within the center of the Earth's shadow may change by 2 orders of magnitude in the visible spectrum. It is therefore evident that dust properties retrieved in the past are eligible candidates for reevaluation to correct a view on the dust trend in the middle atmosphere. Sample calculations are presented for the lunar eclipse observed on 19 January 1954. PMID- 16353811 TI - Microbend-induced mode coupling in a graded-index multimode fiber. AB - We present a theoretical analysis of the mode coupling effects at a microbend along a graded-index (GI) multimode fiber (MMF). By matching the incident and excited mode fields at the microbend, we obtain the coupling coefficients among the guided modes at different microbending conditions. The theoretical results compare well with the experimental results from near-field measurements on a GI MMF subject to microbending. The usefulness of our theory is demonstrated with two applications: (i) an explanation of the operation principle of a wavelength switchable fiber laser that contains a fiber Bragg grating in a GI MMF and (ii) quantification of the function of a microbend-based mode scrambler. PMID- 16353812 TI - Vicarious calibration of the third and fourth Stokes parameters of Windsat measurements. AB - Microwave observations made at the third and fourth Stokes parameters can be used to determine the surface wind direction over oceans. However, due to their smaller amplitudes (less than 3 K at the third Stokes parameter and 1 K at the fourth Stokes parameter), the absolute calibration to these measurements becomes crucial. A new methodology is developed in this study to calibrate the Windsat third and fourth Stokes parameters through tropical rain forest measurements over the Amazon and central Africa. It is found that the Windsat fourth Stokes parameter at 18 GHz has biases of the order of 0.5 K, which could severely affect the wind vector retrievals. PMID- 16353813 TI - In-line type micropulse lidar with an annular beam: experiment. AB - An in-line type compact micropulse lidar (MPL) with an annular beam was developed for low-altitude cloud measurement. An optical circulator and a couple of axicon prisms for an annular beam were installed on the lidar optics. The advantage of using the in-line MPL is its ability to obtain a near-range measurement with a narrow field of view of 0.1 mrad and to obtain a depolarization measurement of the orthogonally polarized echoes caused by ice crystals of a low-altitude cloud. The total insertion loss of the lidar optics was 3 dB. Detectors such as avalance photodiode detectors can be operated in an analog mode near the breakdown voltage because of the high isolation of the optical circulator. The ideal lidar echo variation from the nearest distance was verified by measuring the mountain echoes at various distances. The depolarization measurement of a low-altitude ice cloud was also demonstrated. PMID- 16353814 TI - Two-color laser-induced incandescence (2C-LII) technique for absolute soot volume fraction measurements in flames. AB - A two-color version of the laser-induced incandescence (2C-LII) technique was implemented for measuring absolute soot volume fraction in flames. By using a calibrated tungsten ribbon lamp, soot peak temperatures were measured as a function of fluence at several locations in an ethylene diffusion flame by using a steeply edged laser beam profile. Above a certain fluence threshold, peak temperatures were tightly distributed just above 4000 K independent of the particle size and number density. Radial profiles of soot volume fraction were obtained and compared (not calibrated) with results from the laser extinction technique. Good agreement showed the validity of the 2C-LII technique at a controlled fluence. PMID- 16353815 TI - Imaging through turbulence with a quadrature-phase optical interferometer. AB - We present an improved technique for imaging through turbulence at visible wavelengths using a rotation shearing pupil-plane interferometer, intended for astronomical and terrestrial imaging applications. While previous astronomical rotation shearing interferometers have made only visibility modulus measurements, this interferometer makes four simultaneous measurements on each interferometric baseline, with phase differences of pi/2 between each measurement, allowing complex visibility measurements (modulus and phase) across the entire input pupil in a single exposure. This technique offers excellent wavefront resolution, allowing operation at visible wavelengths on large apertures, is potentially immune to amplitude fluctuations (scintillation), and may offer superior calibration capabilities to other imaging techniques. The interferometer has been tested in the laboratory under weakly aberrating conditions and at Palomar Observatory under ordinary astronomical observing conditions. This research is based partly on observations obtained at the Hale Telescope. PMID- 16353816 TI - Continuous-wave laser performance of Nd:LuVO4 crystal operating at 1.34 microm. AB - A laser-diode-array end-pumped Nd:LuVO4 crystal continuous-wave (cw) laser operating at 1.34 microm has been demonstrated. The maximum cw output power of 1.85 W was obtained at the incident pump power of 17 W for a c-cut 0.5 at. % Nd doped LuVO4 crystal sample, giving the corresponding optical conversion efficiency of 10.88% and a slope efficiency of 13.5%. Laser experiments of Nd3+ concentration of 0.5 and 0.9 at. % a-cut crystal LuVO4 samples were also investigated; due to the strong excited-state absorption of Nd:LuVO4 at 1.34 microm, the output power was limited. PMID- 16353817 TI - Three-dimensional calculation of high-power, annularly distributed, laser-beam induced thermal effects on reflectors and windows. AB - Based on the three-dimensional transient heat conduction equation and the elastic stress-strain equation, the temperature rise, distortion, and equivalent stress distributions of a high-reflectivity silicon reflector and a white bijou window irradiated by a high-power sloped annularly distributed laser beam are simulated using a three-dimensional finite element model (FEM). The effects of laser intensity, output duration, beam obscure ratio, and laser intensity spatial gradient on the results are especially investigated. The effects of mirror and window thermal distortion on laser beam phase aberrations are also evaluated. This noncylindrosymmetric three-dimensional FEM can be used to evaluate high power, high-energy, laser beam-induced thermal effects on optical components. PMID- 16353818 TI - Spatial distribution of power coupling in self-pumped photorefractive reflection gratings. AB - The spatial distribution of the power transfer achieved by contradirectional two beam coupling using self-pumped photorefractive reflection gratings is investigated in two materials with different photorefractive gain coefficients, LiNbO3:Fe and KNbO3:Fe. Incremental portions of the volume grating are erased optically by inducing thin optical damage planes, reducing the overall two-beam coupling efficiency. By monitoring the effect of local grating disruption, the distribution of power transfer is spatially resolved throughout the crystal, and the results are found to be in agreement with our theoretical predictions. PMID- 16353819 TI - Investigation of multiconversion processes in periodically poled LiNbO3-based optical parametric oscillators. AB - The multiconversion processes in optical parametric oscillators based on periodically poled LiNbO3 are investigated. Interpretations based on simultaneous quasi- and birefringent phase matching are presented. Three parametric and three harmonic generation processes in a multigrating periodically poled LiNbO3 crystal were observed. Two of the parametric processes and two of the harmonics were quasi-phase matched, and the other conversion processes were phase matched through birefringence in the crystal. The primary parametric process (omegap --> omegas + omegai) was obtained through first-order quasi-phase matching. The other quasi-phase-matched processes occurred within higher orders. The existence of even-order quasi-phase matching in the crystal is due to other than a 50% duty cycle grating periods. The tuning range for each of the generated waves is obtained and compared with theoretical fittings. PMID- 16353820 TI - In-line type micropulse lidar with an annular beam: theoretical approach. AB - An in-line type micropulse lidar (MPL) with an annular beam was designed and the transmitting and receiving characteristics were analyzed. Because the in-line MPL utilizes a common telescope for a transmitter and a receiver and the annular beam always overlaps with the receiver's field of view (FOV), it can measure near range lidar echoes with a narrow FOV. The transmitting annular beam changes its shape to a nearly nondiffractive beam through propagation. It improves the spatial resolution of the lidar observation. The receiving characteristics showed the ideal lidar echo variation, which was inversely proportional to the square of the distance the beam propagated, even if it was in the near range. PMID- 16353821 TI - Fabrication of subwavelength structure for improvement in light-extraction efficiency of light-emitting devices using a self-assembled pattern of block copolymer. AB - A new fabrication method to improve the optical extraction efficiency of light emitting devices is presented. The morphology of a self-assembled block copolymer was transferred to the surface of a compound semiconductor to achieve a subwavelength columnar structure. The optical extraction efficiency of the substrates with subwavelength columnar structures of 350 nm pillar height, 130 nm diameter, and 180 nm pitch, improved 2.2 times compared to unprocessed substrates. This method does not require expensive exposure lithography tools and is therefore suitable for conventional semiconductor processes. PMID- 16353822 TI - Oceanic inherent optical properties: proposed single laser lidar and retrieval theory. AB - It is suggested that an economical airborne lidar having a single laser can retrieve the three principal inherent optical properties of the ocean. Only three time-resolved backscattering receiver channels are required: (i) elastic (on wavelength), (ii) inelastic (water Raman), and (iii) inelastic [chromophoric dissolved organic matter (CDOM) fluorescence channel to remove the CDOM fluorescence interference from the Raman channel]. PMID- 16353823 TI - Patterns in Mie scattering: evolution when normalized by the Rayleigh cross section. AB - An alternative to using the traditional scattering angle theta to describe light scattering from a uniform dielectric sphere is the dimensionless parameter qR, where R is the radius of the sphere, q = 2k sin(theta/2), and k is the wavenumber of the incident light. Simple patterns appear in the scattered intensity if qR is used in place of theta. These patterns are characterized by the envelopes approximating the scattered intensity distributions and are quantified by the phase-shift parameter rho = 2kR/m - 1/, where m is the real refractive index of the sphere. Here we find new patterns in these envelopes when the scattered intensity is normalized to the Rayleigh differential cross section. Mie scattering is found to be similar to Rayleigh scattering when p < 1 and follows simple patterns for p > 1, which evolve predictably as a function of p. These patterns allow us to present a unifying picture of the evolution of Mie scattering for changes in kR and m. PMID- 16353824 TI - Probiotics-mixed messages. PMID- 16353826 TI - Sexual consequences of prostate cancer treatment. PMID- 16353825 TI - Clarifying the evidence: vitamin E, vitamin A, and folate. PMID- 16353827 TI - A disability paradox. PMID- 16353828 TI - Taking probiotics during pregnancy. Are they useful therapy for mothers and newborns? AB - QUESTION: Recently, several of my pregnant patients have asked me about using probiotics during pregnancy. Is there any evidence that these innocuous bacteria work effectively? ANSWER: An increasing body of evidence suggests that probiotics are effective for treating bacterial vaginosis and allergic reactions. Most probiotics available in Canada, however, are of dubious quality, and, for many claimed indications, there is no proof of effectiveness yet. PMID- 16353829 TI - Use of steroids for erythema multiforme in children. AB - QUESTION: I recently diagnosed an erythema multiforme rash in several patients, two of whom had the major variant, Stevens-Johnson syndrome. Should these patients be managed with corticosteroids? ANSWER: In most cases, mild erythema multiforme is self-limited and resolves in 2 to 4 weeks. Stevens-Johnson syndrome is a serious disease that involves the mucous membranes and lasts up to 6 weeks. There is no indication for using steroids for the mild form. Use of steroids for erythema multiforme major is debatable because no randomized studies clearly indicate which children will benefit from this treatment. PMID- 16353830 TI - Drugs for Alzheimer disease. PMID- 16353831 TI - Probiotics. Some evidence of their effectiveness. AB - OBJECTIVE: To define the term probiotics, to indicate how to identify products that have been proven beneficial, and to assess the quality of evidence regarding probiotics. QUALITY OF EVIDENCE: A few level I studies support the effectiveness of specific probiotics for certain diagnoses. For most so-called probiotics, however, weak or no evidence supports their effectiveness. MAIN MESSAGE: Probiotics are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. Level I evidence supports use of VSL#3 for maintaining remission of inflammatory colitis. Probiotics for treating vaginal infections, Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14, have level I evidence of effectiveness, but are not available in Canada. Specific probiotics taken for certain indications improve health and have few side effects. CONCLUSION: Limited but good evidence supports the role of certain probiotics in medical practice. Because consumer pressure will undoubtedly stimulate further interest in probiotics, family doctors need to be informed about them so they can advise their patients appropriately. PMID- 16353832 TI - Radiotherapy basics for family physicians. Potent tool for symptom relief. AB - OBJECTIVE: To review the use of radiotherapy in cancer care, especially the role of radiation in palliative care. QUALITY OF EVIDENCE: MEDLINE was searched from January 1966 to January 2004. Relevant articles evaluating indications for radiotherapy were reviewed. This paper describes use of radiotherapy based on well established current radiotherapy practices and level I and II evidence. MAIN MESSAGE: A substantial number of patients seeing family physicians develop cancer or die of it. Despite its effectiveness, radiotherapy remains underused in management of cancer patients. Broad uses of radiotherapy in cancer care include curative treatment, palliative symptom control, and management of oncologic emergencies. Radiotherapy can be especially effective for treating common complications of cancer, including painful bone and soft tissue metastases, hemoptysis, dyspnea, dysphagia, brain metastases, and spinal cord compression. CONCLUSION: Radiotherapy is a vital aspect of both curative and palliative cancer treatment. Understanding the basics of radiotherapy can assist family physicians in providing complete care for their cancer patients. PMID- 16353833 TI - Flu shot. PMID- 16353834 TI - Residents' guide to your first locum tenens. What do you have planned after residency? PMID- 16353836 TI - [Gynecology-obstetrics: a changing specialty]. PMID- 16353835 TI - What constitutes clinical evidence? A dynamic approach to clinical diagnosis. AB - The evidence-based diagnostic approach appreciates physicians' inability to eliminate uncertainties in clinical medicine. Likelihood ratios can be a dynamic tool for refining probability in clinical settings. Family physicians can use these parameters for better estimation of the strength of clinical evidence. PMID- 16353837 TI - [The epidemic of cesarean sections: a necessary evil?]. AB - The proportion of cesarean deliveries has raised worldwide, to reach 29% in Switzerland, with important differences between public and private hospitals. There is no simple explanation to this. The emphasis on safety is no excuse, as cesarean sections have short and long-term risks. In this article, we try to understand the reason of this epidemic and we discuss the benefits and risks of cesarean section. This intervention should be performed for medical reasons only. Maternal demand is acceptable, only after a real informed consent. While simplification of surgical techniques and loco-regional anesthesia made cesarean safer, it should not become commonplace. To respect physiology is obstetrician's primary duty, corresponding to the wish of women. Hospitals and clinics have to publish their cesarean section rate. PMID- 16353838 TI - [New options in the treatment of ano-rectal incontinence]. AB - Ano-rectal incontinence is known to affect about 2% of the population. Main risk factors are traumatic delivery and previous anal surgery. All patients should have a trial of conservative treatment. Patients with major external anal sphincter defect have a 70 to 80% improvement of their symptoms after an overlap sphincter repair Unfortunately, these results deteriorate over time. Sacral nerve modulation improves continence and quality of life in 75 to 100% of patients with various aetiologies. In case of idiopathic internal sphincter degeneration, sphincter augmentation with bulking agents seems to be the least expensive option. PMID- 16353839 TI - [Sexual and reproductive health--specific problems in adolescence]. AB - The specificity of adolescent gynecology is uncontested and asks for a specific knowledge and approach. Even if the symptoms remain the same, differential diagnostic and therapeutic approach will be different, depending on the state of development (early puberty, late adolescence), during which they occur. The ignorance of specific pathologies (utero-vaginal malformation, ovarian pathologies related to the menstrual cycle) and therefore inadequate or retarded medical treatment might be the source of unnecessary sequellae. There are now many possibilities to acquire or improve one's knowledge and skills in this particular domain of gynecology. The medico-legal situation as well differs in many points from that of the adult. PMID- 16353840 TI - [Female urinary incontinence: a review]. AB - Female urinary incontinence is a frequent problem. The diagnosis is quite easy, but a precise interview and often an urodynamic exam have to be performed to assess the aetiology. Multiple pathologies cause incontinence. The therapeutic possibilities are various, especially for stress incontinence and urge. Stress, urge and mixed incontinence are the most frequent types. The diagnosis is clinical and in certain circumstances, paraclinic investigations and a specialised consultation are mandatory. Therapeutic success depends from a thorough diagnosis with a treatment based on several parameters, depending on the type of incontinence. The treatment will be medical, physiotherapy or surgery. Success rates varies in function of the type of incontinence. With new surgical techniques, the success rate for stress incontinence is as high as 90%. PMID- 16353841 TI - [Practice guidelines: conservative treatment of fibroids]. AB - The conservative treatment of uterine fibroids is essentially based on symptomatology and patient's choice of treatment. The gynaecologist must develop a clear therapeutic protocol based on clinical examination, available test results and consideration of patient preference. The therapeutic options include close surveillance, hormonal treatments, conservative operative endoscopy and arterial embolization. PMID- 16353842 TI - [Management of postpartum haemorrhage]. AB - Today, postpartum hemorrhage remains a leading cause of maternal morbidity and mortality. Medical treatment, various surgical procedures and/or uterine artery embolisation have considerably reduced the risk of hysterectomy. It is important to identify the different risk factors of hemorrhage after delivery and to take the precautions to avoid it. A clear strategy defined by the obstetrical team is essential to decrease the delay in the management of this complication in order to increase the chances of a successful treatment. PMID- 16353843 TI - [Sleep and normality: usefulness for the practitioner]. AB - About fifty percent of patients consulting their primary care physician complain of sleep disorders. Clinicians have to distinguish normal from abnormal sleep. For example, sleep normality is usually characterised by an apnea/hypopnea index < 5 h(-1) in absence of sleep symptoms. However such an index varies in relation with age, gender, body mass index, measurements methods In daily clinical practice, clinicians should look for sleep symptoms to define an abnormal sleep. More studies are needed to better define which patient will really benefit from a specific sleep therapy. PMID- 16353844 TI - [Agenda for the future: a roadmap of ambulatory pediatrics]. PMID- 16353845 TI - [Outside of science, no salvation?]. PMID- 16353846 TI - [The incomprehensible message from Parma]. PMID- 16353847 TI - [Tobacco smoke progressively moves away from the public places in the European Union]. PMID- 16353848 TI - [The new provocation of Professor Claude Sureau]. PMID- 16353849 TI - [The (too) slow agony of poliovirus]. PMID- 16353850 TI - [Pastiche smoking]. PMID- 16353851 TI - Mapping therapy for sentence production impairments in nonfluent aphasia. AB - This study investigated a new treatment in which sentence production abilities were trained in a small group of individuals and nonfluent aphasia. It was based upon a mapping therapy approach which holds that sentence production and comprehension impairments are due to difficulties in mapping between the meaning form (thematic roles) and the syntactic form of sentences. We trained production of both canonical and noncanonical reversible sentences. Three patients received treatment and two served as control participants. Patients who received treatment demonstrated acquisition of all trained sentence structures. They also demonstrated across-task generalisation of treated and some untreated sentence structures on two tasks of constrained sentence production, and showed some improvements on a narrative task. One control participant improved on some of these measures and the other did not. There was no noted improvement in sentence comprehension abilities following treatment. Results are discussed with reference to the heterogeneity of underlying impairments in sentence production impairments in nonfluent patients, and the possible mechanisms by which improvement in sentence production might have been achieved in treatment. PMID- 16353852 TI - Design your own memory strategies! Self-generated strategy training versus mnemonic training in old age: an 8-month follow-up. AB - We examined maintenance effects following training in a self-generated strategy versus a mnemonic strategy, designed to improve recall of four-digit numbers in older adults. This was accomplished by reassessing participants from a previous study eight months after completion of training. A control group that received no training was also included. The results showed that both training groups maintained training-related gains in number recall performance eight months later in a criterion task providing cognitive support. However, when no support was provided, number recall performance for the mnemonic group dropped slightly whereas performance for the self-generated strategy group improved. The control group maintained performance across assessments. These data indicate the numerical memory training may result in long-term effects in older adults and that, under some conditions, practising one's own memory strategies may have advantages over mnemonic training in old age. PMID- 16353853 TI - The sensitivity and specificity of the Middlesex Elderly Assessment of Mental State (MEAMS) for detecting cognitive impairment after stroke. AB - The aim of the study was to assess the sensitivity and specificity of the MEAMS (Golding, 1989) for detecting cognitive impairment after stroke. Stroke patients admitted to hospital received a cognitive screening assessment, the MEAMS, and a detailed cognitive assessment. The information obtained from the detailed assessment was summarised in a structured written report. From the conclusions in these reports, patients were classified as "impaired" or "not impaired" in perception, memory, executive function and language. The sensitivity and specificity of the MEAMS subtests and the overall number of tests passed were determined in relation to the presence of impairment, as given in the overall conclusion of the written reports. There were 30 stroke patients, aged 58 to 92 (mean 75.80, SD 7.94) years. Of these, 17 were men and 13 were women. The sensitivity of the MEAMS subtests ranged from 11% to 100% and the specificity ranged from 69% to 100%. The sensitivity of the overall MEAMS score was 52% and the specificity was 100%, using a cut-off score of 3 or more fails to indicate impairment. Three subtests, Orientation, Naming and Unusual views had 81% sensitivity and 50% specificity for detecting problems in language, perception or memory. The MEAMS was not a sensitive screen for overall cognitive impairment or for memory, perceptual, language, or executive function problems after stroke, but it was specific. Although screening for cognitive impairment is important, the MEAMS is not recommended as the sole method, as it produces an unacceptably high false negative rate. Three subtests (Orientation, Naming and Unusual views) had 81% sensitivity and 50% specificity for detecting cognitive problems in language, perception or memory after stroke. PMID- 16353854 TI - NART prediction and impairment in neurological patients. AB - Crawford, Allan, Cochrane, and Parker (1990) used demographic variables in a multiple regression equation to predict NART error scores in a general population sample. The present study examined the applicability of Crawford's equation to neurological patients. Using this equation and the authors' suggestion that NART performance can be impaired in neurological patients, the data were also examined in participants with diffuse cortical damage. Participants (n = 175) were patients referred for clinical neuropsychological evaluation in north-east England. The findings confirmed the usefulness of Crawford's equation: Participants showing significantly higher obtained than predicted NART error scores (identified using Crawford's equation), were categorised as NART-impaired. Analyses of NART-impaired and NART-unimpaired patient subgroups showing diffuse cortical damage indicated the former had a significantly higher mean NART error score and mean discrepancy between NART-predicted and observed IQs. However, a significantly lower mean verbal (VIQ) IQ score was also noted in the NART impaired participants, suggesting that the risk of an impaired NART performance is greater in a diffusely damaged patient with a lower premorbid VIQ. PMID- 16353855 TI - Filling the quality gap: physican-hospital QIs. PMID- 16353856 TI - Improving bariatric services marketing at your hospital. PMID- 16353857 TI - Neoadjuvant chemotherapy for oesophageal cancer: the need for accurate response prediction and evaluation. AB - Primary surgical resection for locally advanced oesophageal cancer is associated with systemic failure and poor survival due to presence of micrometastatic disease at the time of diagnosis. Neoadjuvant chemotherapy prior to surgical resection aims to downstage these locally advanced tumours. A review of reported randomised controlled trials has shown only one sufficiently powered trial with a survival advantage for cisplatin-based chemotherapy. Published meta-analyses of neoadjuvant chemotherapy trials have shown little or no overall survival benefit. A subgroup of patients with biologically favourable tumours who respond to this treatment have been consistently shown to have a survival advantage. These patients need to be differentiated from non-responders preferably at an early stage of this potentially toxic treatment. Current clinical, endoscopic and radiological methods of response evaluation are all unreliable. Response evaluation with 18FDG-PET has been shown to accurately assess the pathological response and also to predict the risk of local recurrence and overall survival. The development of integrated PET/CT imaging may enhance the accuracy of this response evaluation. In the future, molecular markers of response prediction prior to initiation of treatment may allow the development of individualised treatment strategies. New emerging chemotherapeutic agents may prove to be more effective in eradicating micrometastatic disease. PMID- 16353858 TI - SELDI-TOF proteomic analysis and cancer detection. AB - BACKGROUND: In recent years proteomic approaches have been widely used to diagnose disease and the new technology of surface enhanced laser-desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS) is very promising. METHODS: A review of English language literature was undertaken using Internet databases such as PubMed and Medline for studies using proteomic technologies for the early detection of cancer. RESULTS: SELDI-TOF is an array based mass spectrometric method in which proteins of interest are selectively absorbed onto a chemically modified surface and the mass and amount of each protein is measured by irradiating the surface with a laser and measuring the time-of-flight. The technology has revealed a large number of previously uncharacterized biomarkers for a wide variety of cancers. Its versatility has also been demonstrated by its application as a tool for mining a wide variety of biological tissues and fluids. CONCLUSIONS: The majority of these studies have discriminated between diseased and healthy controls with a high degree of sensitivity and specificity. The reproducibility, standardization and feasibility of this technology need to be addressed before these proteomic approaches to the discovery of novel, highly sensitive diagnostic tools can become routine clinical care. PMID- 16353859 TI - Children's surgery performed by adult general surgeons in Scotland: the present and future. AB - OBJECTIVE: The aim of this study was to determine the extent and the future of paediatric surgery performed in Scotland outside of the designated surgical paediatric centres. MATERIALS AND METHODS: An anonymous questionnaire was sent to all 111 Scottish members of the Association of Surgeons of Great Britain and Ireland. There was a response rate of 69%. RESULTS: Overall, 45% of responders operated on children. This was independent of the surgeon's age but was related to the type of hospital that the surgeon worked in. Eighty-four per cent of responders had a lower age limit under which they would not operate and 94% stated that there were specific circumstances where they would not operate. A mean of 18.5 elective procedures (range 0-250, median two) and six emergency procedures (range 0-30, median five) were carried out by each surgeon operating on children under the age of five per annum. Only 13% of responders thought that their successor would operate on children. CONCLUSIONS: Non-specialist paediatric surgery in Scotland is currently provided by a significant number of surgeons whose successors will not continue to provide a comparative paediatric service. This has implications for local provision of care, emergency management and capacity of existing children's hospitals in the future. PMID- 16353860 TI - Paralysis after aortic surgery: in search of lost cord function. AB - During the early 1980s, the rate of paralysis after aortic surgery was as high as 41% in patients for the most complex thoracoabdominal aortic operations. After comparing human and chacma baboon (papio ursinus) spinal cord vascular anatomy, an animal model was established to study the pathophysiology of aortic cross clamping and the aetiology of the paralysis. Techniques, including motor evoked responses for monitoring spinal cord function, were developed that were tried in humans and later culminated in prospective and randomized studies. These established that the following were protective: combining cerebrospinal fluid with intrathecal papaverine; cooling systemically to moderate or profound hypothermia; minimizing intercostal ischaemia time; using a sequential segmental repair approach; re-attaching all patent and segmental intercostal arteries below T8 for descending thoracic aortic repair and from T6 to L2 for thoracoabdominal repairs; continuing cerebrospinal fluid drainage for at least two days and maintaining patients hypertensive after surgery. The net result has been that, in two of our recent series, the risk of permanent paralysis has been reduced to between 3.1% and 3.8%. PMID- 16353861 TI - My friend Dr Knox: a pupil writes about the anatomist. AB - Thomas Giordani Wright, a medical apprentice in Newcastle upon Tyne, attended Dr Robert Knox's anatomy classes in Edinburgh between November 1825 and April 1826, only two years before Burke and Hare began murdering people and selling the bodies of their victims to Knox's anatomy school. In March 1829, soon after the crimes came to light and Burke had been found guilty of murder and executed, Wright commented on the case in his diary, describing the practices in the dissection rooms and giving his views on Robert Knox, the teacher who had become his friend. PMID- 16353862 TI - Something not quite right: Gardner syndrome diagnosed by multiple cutaneous lesions and genetic testing. AB - Gardner syndrome is a variant of familial adenomatous polyposis characterized by intestinal adenomatous polyps, which can progress to adenocarcinoma, and a variety of extraintestinal manifestations, including skin cysts, osteomas, soft tissue fibrous tumours and a characteristic ocular lesion. The extraintestinal manifestations are often the presenting feature but are usually not sufficiently characteristic on their own to trigger recognition of the syndrome. We report a case of a 17-year-old female who had been treated by a number of specialists over a 13-year period for a variety of cutaneous lesions without a hereditary condition being suspected. Gardner syndrome was considered only after excision of subcutaneous fibrous tumours from the mastoid region and paraspinal area and was confirmed by genetic testing in spite of the patient's refusal to undergo colonic endoscopic examination. Subsequent resection revealed approximately 70 adenomatous colonic polyps in the colon and rectum but no invasive tumour, highlighting the benefits of genetic testing in treatment planning. PMID- 16353864 TI - Re: an audit of ultrasound-assisted catheter insertion in patients receiving chemotherapy. Surgeon 2005;3(1):32-35. PMID- 16353863 TI - Transanal endoscopic microsurgical repair of iatrogenic recto-urethral fistula. AB - BACKGROUND: Recto-urethral fistula formation following radical prostatectomy is an uncommon but potentially devastating event. Traditional surgery for such fistulae is technically demanding, jeopardizes continence and usually necessitates a diverting colostomy. We present the case of an iatrogenic fistula treated by a transanal endoscopic microsurgical approach, without recourse to a stoma. METHOD: A 71-year-old man had recently undergone a radical prostatectomy, complicated by significant intra-operative haemorrhage. He subsequently developed a recto-urethral fistula, confirmed clinically and endoscopically. Due to his bleeding diathesis, he was considered for, and underwent, a transanal endoscopic microsurgical (TEMS) repair. CONCLUSION: TEMS is a safe and reliable minimally invasive surgical technique for the treatment of this difficult condition. PMID- 16353865 TI - [Organization of medicinal service in local military conflicts of our times]. AB - In modern conditions the increasing frequency of local conflicts makes it necessary to pay special attention to some problems of medical support system taking into account the experience of previous military campaigns. Among these problems are the following: the realization of principle of territorial supply; the creation of distinct system of interdepartmental interaction in conflict zone; the improvement of individual and group medical equipment; as well as the development of standards of drug and medical material supply basing on the common approaches to treatment and care rendered to the patients and casualties. PMID- 16353866 TI - [Methodical approaches to distribution of population medical support means among the regions in conditions of extreme situations]. PMID- 16353867 TI - [Xenon anesthesia of the abdominal surgery]. PMID- 16353868 TI - [The influence of induced severe myocardial ischemia on coronary disease course]. AB - The aim is to study influence of short-term episodes ischemic of myocardium in a combination with intravenous introduction of creatine phosphate on current of ischemic heart disease at the patients with distal stenoses of coronary arteries. We survey 86 patients, which were divided into two groups: basic, which are receiving neoton in a combination with ischemic training and control who are receiving standard antianginal therapy. The research proceeded within 5 years. The authentic improvement of tolerance to physical effort, quality of life, decrease anginal attacks and normalization systolic and diastolic cardiac function of the basic group is marked. PMID- 16353869 TI - [Clinical, laboratory and instrumental precursors of ciliary arrhythmia in the patients with diffuse toxic goiter]. PMID- 16353870 TI - [Exercise therapy for ureterolithiasis after remote stroke-wave and contact lithotripsy]. PMID- 16353871 TI - [The main directions of "restorative ophthalmology" in conditions of multi purpose rehabilitation center]. AB - The authors have developed the 4-level system for restoration of operator's professional vision basing on the principles of conception's project of health protection in Russian Federation. On the hole the complex system of sanitary measures includes the following directions: the wide use of self-prophylaxis and correction methods of visual disorders, the purposeful use of effective means of visual profile ("operative correction") directly in the operator's working place, as well as the conduction of complex stimulation of vision organ on the basis of specialized room ("long-term correction"). PMID- 16353872 TI - [The surgical approaches in hysterectomy]. PMID- 16353873 TI - [Experience of prophylaxis of extra-hospital pneumonia with the use of vaccine "Pneumo-23" conducted in the military collectives]. PMID- 16353874 TI - [AIDS-associated pneumocystic pneumonia]. PMID- 16353875 TI - [The ways for improvement of infusion solution preparation in field conditions]. AB - To solve the problem of more effective preparation of infusion solutions in field conditions is impossible without conduction of research and experimental-and designers' work. The main direction of such work is creation of modern means of infusion solution packing with the use of polymeric materials (containers); the sterilization complex with filtering and aseptic dosed pouring of infusion solutions into containers; the water-obtaining installation for injections in field conditions from natural sources. Besides a set of technical means (the complex for infusion solution preparation in polymeric containers, the water obtaining installation for injections in field conditions, etc.) should be created. Also it is necessary to have the mobile technical means (the field laboratory of infusion solutions) that allows conducting all measures for mass preparation of infusion solutions in polymeric containers under field conditions. PMID- 16353876 TI - [Methodics of analysis of specialized equipment competitiveness for conduction of forensic examinations]. PMID- 16353877 TI - [Half-centenary of military field therapy department of Kirov Military Medical Academy]. PMID- 16353879 TI - [Surgical treatment of Crohn's disease]. AB - OBJECTIVE: The experience with the surgical treatment of Crohn's Disease is analyzed in order to determine the indications for surgery, surgical procedures, postoperative complications and associated risk factors, and recurrence rates. MATERIAL AND METHODS: Retrospective analysis of patients operated upon for Crohn's Disease between 1979 and 1997. Risk factors for operative morbidity were searched, and actuarial curves for recurrence of disease were constructed. RESULTS: There were 34 patients with a mean age of 46 years. All patients were operated upon because of complications; the most frequent ones were stenosis and obstruction. The affected patterns were ileocecal in 53%, colitis in 35% and small bowel in 12%. Segmental resections were the most frequent ones, mainly ileo cecal. Operative morbidity was 32% and mortality 6%. The presence of colonic affection and the necessity of its resection was a higher risk for complications (P< 0.05). Actuarial recurrence was 39% at 107 months. CONCLUSIONS: Surgery for Crohn's disease was mainly indicated in complications, presented high rates of morbidity associated with colonic involvement and resections, and there were high rates of recurrence at long-term. PMID- 16353880 TI - [Clinical characteristics and follow-up of patients with AIDS and acute abdominal pain]. AB - Abdominal symptoms frequently affect patients with AIDS. Acute abdominal pain is a diagnostic challenge that may require elective or urgent surgical treatment, although information about the latter is scarce. In this study we analyzed the clinical findings and follow-up of acute abdominal pain complicating patients with AIDS. In a two-year period, we collected several variables from patients with AIDS and acute abdominal pain: demographic, laboratory, clinical symptoms, initial diagnosis, surgical findings, post-surgical and histopathological diagnosis and post-surgical complications. From 232 hospitalized patients, 34 had acute abdominal pain: 32 male and 2 women (median age = 32 years; range 26 to 58 years). Twenty-two patients required surgical treatment. Eight patients had a post-surgical complication; in five of them, six surgical re-interventions were performed. Three deaths occurred in the 30-day period after surgery. Survival for patients conservatively treated was 4 months (1 to 17 months), vs. 6.5 months (1 to 20 months) in the surgically treated group. Physicians should be aware about the several diagnostic possibilities of acute abdominal pain complicating patients with AIDS. Delay of surgery in these patients may be lethal. Surgery has an important role in the integral treatment of patients with AIDS. PMID- 16353881 TI - [Efficacy of prednisolone and rimexolone in HLA-B27 positive patients with acute anterior uveitis]. AB - PURPOSE: To compare the efficacy and safety of prednisolone acetate 1% vs. rimexolone 1% ophthalmic suspension in the treatment of acute anterior uveitis (AAU) in HLA-B27+ patients. METHODS: Sixty-eight AAU HLA-B27+ patients were randomly selected for treatment with prednisolone acetate 1% or Rimexolone 1%. All patients showed mild to moderate anterior chamber inflammation. This was a prospective, randomized, double blind, clinical trial. RESULTS: There was no statistically significant difference between both groups when anterior chamber cells were measured. In the rimexolonae group, flare diminished since the first week. In both groups the intraocular pressure (IOP) raised since the first week, the increase was highly significant in the rimexolone group. Final intraocular pressure was higher in the prednisolone group. CONCLUSION: Rimexolone 1% is as effective as prednisolone acetate 1% in the treatment of mild to moderate AAU HLA B27+. IOP increased in both groups, but this variation was not clinically significant. PMID- 16353883 TI - [Distraction osteogenesis as treatment for craniofacial deformities]. AB - Distraction osteogenesis is a surgical technique for treating different craniofacial deformities. The sophisticated clinical expression of craniosynosotosis in the face, as well as in the skull can be corrected using different osteotomies, in which we can include the frontal bone, orbits, the malar bones and the maxillae. Once the osteotomy has been completed, we can use the distraction devices to achieve the planned bone advancement. This procedure can be also performed with endoscopic techniques producing less morbidity, with minimal complications, simultaneously producing a highly satisfactory correction of the frontal deformity, the exorbitismus, the malar and the mid-face retrusion in these patients. As the distraction procedure is gradual, soft tissues resistance against the advancement can be easily superated achieving larger antero-posterior bone advancement that can vary between 16 to 32 mm in the frontal area and between 9 to 16 mm in the maxillae. The new position of the bones are maintained in place for new bone formation areas over the osteotomy lines. The aesthetic and functional changes after the procedure have been highly satisfactory in these patients. PMID- 16353882 TI - [Stereotactic radiosurgery with linear accelerator (LINAC). Report of the technical experience in 100 cases treated at the Mexico General Hospital]. AB - Stereotactic radiosurgery consists of the release and high dose radiation inside the intracraneal, small, well defined and stereotactic localized injury. OBJECTIVE: In this study we describe the technical aspects, clinical results and dose radiation levels used in 100 patients treated with LINAC. METHODS: One hundred patients received treatment with LINAC stereotactic radiosurgery, 34 AVM, 22 meningiomas, 18 astrocytomas, 11 pituitary adenomas, 5 cavernous malformations, 3 acoustic neuromas, 3 craniopharyngiomas, 2 cases of epilepsy, one brain metastasis and one ependymoma. The radiation dose changed in each case. Studied variables: response of the injury, toxicity, in target dose, number of isocenters, number of treatment arcs, number o fcollimators, dose of risk organ, homogeneity of target, margin of error, target, diameter of collimator and location of target. We applied the chi2 test with statistical significance of p < 0.05. RESULTS: The median follow-up was 17.7 months. We had 3 cases of minor toxicity. We confirmed that for a greater target volume, greater number of treatment arcs were necessary, with greater margins of errors and greater number of isocenters. CONCLUSIONS: Radiosurgery is a safe and reliable technique to treat benign and malignant intracraneal lesions. PMID- 16353884 TI - [Mortality in cardiogenic shock]. AB - OBJECTIVE: Cardiogenic shock (CS) is one of principal causes of mortality after an acute myocardial infarction (MI). The objective of this study was to determine the principal causes that contribute to an increase in mortality in CS. METHODS: We studied 155 consecutive patients with CS admitted to the Coronary Care Unit of the Instituto Nacional de Cardiologia Ignacio Chavez from 1990-2002. RESULTS: Patients older than 60 years with MI and diabetes mellitus presented a higher cardiovascular mortality (p<0.001). Percutaneous coronary intervention (PCI) procedures decreased the cardiovascular mortaly in CS as compared to those patients not submmitted to PCI (59% vs. 98%, p<0.001). CONCLUSIONS: Mortality due to CS is still very high (80%). Previous MI and diabetes favor short-term mortality and the use of PCI suggests a clinical favourable trend in the reduction of mortality due to CS. PCI appears to be the most appropriate reperfusion procedure for treating CS. PMID- 16353885 TI - [Consistency of HIV/AIDS transmission categories reported by the epidemiological surveillance of Chiapas State, Mexico: a problem of method]. AB - OBJECTIVE: To compare the mechanisms of HIV/AIDS transmission reported by the epidemiological surveillance system information of the Chiapas State and those referred by HIV positive men interviewed in the Soconusco's region from the same state. METHODS: Databases containing information about the HIV/AIDS cases registered in the State of Chiapas and in the Soconusco's region were used. Additionally, 32 men previously detected as HIVpositive by the regional surveillance system answered a questionnaire about their sexual behavior and some socio-demographic characteristics. All participants provided a blood sample to identify antibodies against HIVusing two screening tests, ELISA and microagglutination, and one confirmatory test by Western blot. RESULTS: While the heterosexual transmission was predominant among HIV/AIDS cases reported by the epidemiological surveillance of Chiapas State and the Soconusco's (50.4% and 69. 40%, respectively), heterosexual transmission among the interviewed individuals was 36.7%. CONCLUSIONS: Contrary to the proposal about a process of "heterosexualization" of HIV/AIDS epidemic in Chiapas State and particularly in the Soconusco's region, the present study suggest that homosexual and bisexual behavior continue to be the main routes of HIV transmission among men. These results also suggest the presence of a methodological limitation in the evaluation of sexual behavior among HIV-seropositive individuals in the region. A gender analysis about masculinity, would allow a better understanding ofthe social and cultural aspects of sexuality that are not evident when only the sexual practices are evaluated. PMID- 16353886 TI - [Metabolism of cholesterol and fatty acids in nephrotic syndrome and its regulation by sterol regulatory element binding proteins (SREBP's). Effect of soy protein consumption]. AB - Hyperlipidemia occurs during nephrotic syndrome (NS). It is known that cholesterol and fatty acid biosynthesis is controlled by the transcription factors sterol regulatory element binding proteins (SREBPs). Soy protein consumption reduces the concentration of these lipids, although its mechanism of action is not well known. The aim of the present study was to establish whether soy protein consumption reduces cholesterol and triglycerides levels by regulating of SREBPs. Male Wistar rats with experimental NS were studied for 64 days. The results showed that rats fed with soy protein had significantly lower plasma cholesterol and triglyceride concentrations as well as proteinuria than rats fed with casein diet. These decrements were associated with a decrease in the expression of SREBP-1 and fatty acid biosynthetic enzymes. In addition, Western blot analysis revealed that in nuclear extracts from hepatocytes of rats fed with soy protein, there was a lower concentration of SREBP-1 than in rats fed with casein. The results of this study indicate that consumption of a soy protein diet has beneficial effects on nephrotic syndrome. PMID- 16353887 TI - [Intestinal trichobezoar as a cause of small bowel obstruction. Case report and review of the literature]. AB - BACKGROUND: The trichobezoars are conglomerates of hair ingested by a person. They are located in the stomach, although they can extend also to the small bowel. They are common in children and adolescents, and 90% of the patients are women. An occasional complication is mechanical obstruction, usually at the gastric level or in the small bowel. It is extremely rare to find trichobezoars in the small bowel without a gastric component. CASE REPORT: A 18 year-old female patient was admitted for small bowel obstruction. Three weeks before she had underwent appendectomy. Laparotomy was performed identifying three trichobezoars occluding the jejune, without evidence of gastric bezoars. The bezoars were extracted by enterotomy. No post-operatory complications were presented. CONCLUSIONS: Trichobezoars are a rare entity that may be complicated by bowel obstruction. The discovery of trichobezoars in the bowel may be due to primary formation, which is improbable, or to fragmentation in the stomach followed by migration of the fragments, which in the case presented could be favored by the patient's previous operation. PMID- 16353888 TI - [Appendicitis in a two-months old infant with a peritoneovaginal duct]. AB - Acute appendicitis in infants is a very unusual disease, but associated to structural defects like patent peritoneovaginal duct, Meckel's diverticulum, clubfeet, and Moebius syndrome is extremely rare. Case report. A male of two months-old with this association is presented. Left inguinal swelling and acute abdomen syndrome were identified. In laparotomy, iguinal defect and a gangrenous appendicitis were observed. Appendectomy was performed and postoperative recovery was unevenful. PMID- 16353889 TI - [Diagnostic radiology in the technological era. Comparison between two models]. AB - Diagnostic Radiology is a medical specialty that has played a dynamic role in the medical arena during the last three decades of the XX century. As a part of the evolution of this specialty, medicine and society at large have received diverse benefits in areas of diagnosis and for the decision making process in the clinical practice. Modern radiology has provided numerous advances and refined tools that give more convenience speed and precise diagnosis in the current medical practice. At the present time, these technologies are part of the standard of care in the US and other industrialized countries, representing the ultimate desire or goal to reach for other healthcare systems in developing countries as well. Unfortunately, many of the clinical skills in the health care personal have not evolved at the same pace of those modern technologies, conditioning a reduction in the performance of sensitive areas to the patient's interest, such as the caring, compassion and quality of health care. It is unquestionable the value and benefits that technologies such as CAT and MRI scans have brought to the standard practice in the recent time. Nonetheless, it is desirable to maintain the highest possible level of clinical skills despite the extended use of modern technologies by the health personnel, without compromising the quality of care. Yet is pending the future role and rational use for those technologies in radiology, especially in times when delivering good, reliable health care have become a priority for many health institutions worldwide. This approach would be the ultimate goal to reach in times and circumstances where health for all is the highest precious value to obtain at global level. PMID- 16353890 TI - [An adult woman with abdominal pain and distension, nausea and vomit]. PMID- 16353891 TI - [On the centennial of hormones. A tribute to Ernest H. Starling and William M. Bayliss]. AB - Ernest H. Starling introduced the term hormone 100 years ago in his Croonian Lectures to the Royal College of Physicians in June 1905. It was demonstrated for the first time that one part of the body could influence the function of another distant part. Starling for the first time suggested the word hormone. This review attempts to trace the development of studies in endocrinology, beginning in the middle of the nineteenth century. Starling discovered secretin, the first hormone, in collaboration with William M. Bayliss, and they introduced the hormone concept with recognition of chemical regulation. Thus the name hormone sparked multidisciplinary research in endocrinology and molecular biology, which shed light on the chemical communication within the organism. PMID- 16353893 TI - [Genes and malaria]. PMID- 16353892 TI - [Lissencephaly]. PMID- 16353894 TI - [Risk factors for students desertion from the UASLP School of Medicine, Mexico]. AB - OBJECTIVE: To obtain the profile of students that deserted from the Faculty of Medicine of the Autonomous University of San Luis Potosi, Mexico. DESIGN: Cases and controls nested in a cohort. METHODS: All students that voluntarily deserted between 1992 and 2002 were consulted. Each student was compared in an aleatory form with a regular student and a proper questionnaire was applied. RESULTS: The significantly associated factors to abandon the Faculty of Medicine were: high school of origin (OR=2.43), extra-ordinary exam (OR=3.13), and lack of vocation (OR=2.41). CONCLUSION: The subjacent factors for not deserting from the Faculty of Medicine were: study habits, capacity for sustained effort, and tolerance to frustration. PMID- 16353895 TI - [A Panglossian view of adipocentrism]. PMID- 16353896 TI - [Radiation diagnosis of tumors at four sites (the lung, breast, stomach, and large bowel) in the organizational and methodological standpoints of practical public health of the Russian Federation]. AB - The basic idea of the paper is to put forth today's vision of detecting tumors at four sites (cancer of the lung, breast, stomach, and large bowel) at the level of municipal and regional public health systems. Based on their many years' experience in "contacting" this pathology, the authors characterize the role and significance of basic radiation techniques applied to each of these four tumors, which should be used in their diagnosis. The authors also underline the need for reasonably certifying radiation diagnostic apparatuses for municipal and regional public health systems, which would efficiently combine their diagnostic and economic profitability, which will be able to avoid materializing the potential of expensive radiation equipment by just 15-20% of its built-in efficiency. By taking into account the present epidemiological situation with each of the tumors in question and scientific-and-technological achievements of current diagnostic equipment, the authors underline the need for correcting diagnostic approaches applied in their detection. The paper also presents the opinion of the authors as to selective screening. They are sure that it is impossible without its use to gain even some moderate improvement of the results of diagnosis and hence those of treatment of tumors at four sites, which are all responsible for 50% of all malignancy. The idea that it is necessary to actively return radiation studies to the diagnosis of cancer of the lung, breast, stomach, and large bowel runs throughout the paper. As applied to tumors at each of these sites, the authors express their opinion as to their radiation diagnosis. Naturally, they could not ignore the fact that the authorities of medical industry in our country should take an active part in the solution of organizational problems touched upon in the paper. PMID- 16353897 TI - [X-ray manifestations of contractures of the temporomandibular articulation]. AB - This paper describes the X-ray manifestations of chewer contracture that impairs the movement of the lower jaw and the function of the temporomandibular articulation. PMID- 16353898 TI - [Peripheral carcinoma of the lung in magnetic resonance imaging]. AB - This study shows magnetic resonance imaging (MRI) to be a technique in the comprehensive diagnosis of peripheral carcinoma of the lung and in the determination of the extent of a tumor process in the chest. The paper specifies and systematizes the MRI signs of peripheral carcinoma of the lung, the signs of tumor invasion into the pleura, chest, and mediastinal structures. It also analyzes additional capacities of contrast enhancement. PMID- 16353899 TI - [Magnetic resonance imaging in the diagnosis of nonorganic hydatid disease]. AB - This article evaluates MRI diagnostic value in discovering of the non-organic hydatid disease. MRI data of 21 patients, suffering from parasite pathology of liver (n = 12), liver and peritoneum cavity (n = 2), liver and retroperitoneal space (n = 2), liver and thigh's muscles (n = 1), peritoneum cavity (n = 2), retroperitoneal space (n = 1), spine and paravertebral area (n = 1) were analyzed. Based on histopathological results, features of unusually localized hydatid cysts (HC) MRI- semiotics are described in detail and compared with liver echinococcosis. MRI technique for identification of some hydatid cyst's structures is shown. The authors discuss the MRI reliability in differential diagnosis of non-organic HC and several disorders (non-parasite congenital and acquired cysts, hematoma, abscess, metastasis) of the same anatomical region. They underline some MRI advantages in GD disclosing comparing with ultrasonography and computed tomography. However, serological tests are needed for diagnosis verification. The authors also postulate the importance of clinical data being taken into account for radiological conclusion. PMID- 16353900 TI - [MALTlymphoma of the lung: a review of literature and case report]. PMID- 16353901 TI - [Spiral computed tomography in the differential diagnosis of renal angiomyolipoma with the minimum amount of fatty tissue and renal-cell carcinoma with fatty inclusions]. PMID- 16353902 TI - [Contrast-enhanced magnetic resonance imaging in the diagnosis of urinary tract disease]. PMID- 16353903 TI - [Radiation diagnosis in cardiology (according to the proceedings of the Russian National Congress of Cardiology "Russian Cardiology: from the Center to Regions", Tomsk October 12-14, 2004). Part 2]. PMID- 16353904 TI - [Economic profitability: current aspects of X-ray contrast diagnosis in urology]. AB - A diversity of X-ray contrast diagnostic techniques used in urology provides a way of using different groups of X-ray contrast materials (XCMs) in accordance of the set tasks. Many years' experience in handling water-soluble triiodinated XCMs while examining the kidneys and urinary tract provides evidence for their diagnostic effectiveness and safety. The new Triombrast (OAO "Farmac", Kiev) synthesized on the basis of the Spanish substance (Justesa Imagen SA) has successfully undergone clinical trials and meets the high requirements made by the specialized urological facilities is economically profitable. The paper has been prepared, by using the reports on the results of preclinical studies of the agent and the experience of its use in the clinic of the Research Institute of Urology and City Clinical Hospital No. 47, Moscow. PMID- 16353905 TI - Pharmacodynamic parameter estimation: population size versus number of samples. AB - The purpose of this study was to evaluate the effects of population size, number of samples per individual, and level of interindividual variability (IIV) on the accuracy and precision of pharmacodynamic (PD) parameter estimates. Response data were simulated from concentration input data for an inhibitory sigmoid drug efficacy (E(max)) model using Nonlinear Mixed Effect Modeling, version 5 (NONMEM). Seven designs were investigated using different concentration sampling windows ranging from 0 to 3 EC(50) (EC(50) is the drug concentration at 50% of the E(max)) units. The response data were used to estimate the PD and variability parameters in NONMEM. The accuracy and precision of parameter estimates after 100 replications were assessed using the mean and SD of percent prediction error, respectively. Four samples per individual were sufficient to provide accurate and precise estimate of almost all of the PD and variability parameters, with 100 individuals and IIV of 30%. Reduction of sample size resulted in imprecise estimates of the variability parameters; however, the PD parameter estimates were still precise. At 45% IIV, designs with 5 samples per individual behaved better than those designs with 4 samples per individual. For a moderately variable drug with a high Hill coefficient, sampling from the 0.1 to 1, 1 to 2, 2 to 2.5, and 2.5 to 3 EC(50) window is sufficient to estimate the parameters reliably in a PD study. PMID- 16353906 TI - A guide for reporting the results of population pharmacokinetic analyses: a Swedish perspective. AB - Population pharmacokinetic analyses are frequently part of regulatory submissions and are mainly used to provide information on special populations (effects of age, renal impairment, etc) and drug-drug interactions. A varying standard of population analysis reports has been received at the Medical Products Agency in Sweden, some very good and some unassessable. In the latter case, it may be that it is a report of an inadequate analysis or may be a report of a perfectly acceptable analysis, but too little detail has been provided in the report for the conclusions reached to be properly assessed. A sufficient level of detail must be present in these reports in order for them to be assessable and to allow the conclusions reached to be incorporated into the summary of product characteristics. The report should specify the goal(s) of the analysis, describe in detail the origin and nature of the data, clearly describe the model-building process, include a range of goodness of fit (GOF) plots to support decisions made during the model-building process, and demonstrate that the final model is a good description of the data. The use of color in GOF plots is encouraged so that key features are easily visible. Covariate effects in the final model should be clearly presented and their clinical relevance discussed. In the case of many covariates in the final model, it may be useful to perform some simulations to illustrate the effect of various covariate combinations for a series of different "typical" subjects. PMID- 16353907 TI - Preface: challenges and issues in veterinary pharmacology and animal health - 2005. PMID- 16353908 TI - Postexposure management and treatment of anthrax in dogs--executive councils of the American Academy of Veterinary Pharmacology and Therapeutics and the American College of Veterinary Clinical Pharmacology. AB - Dogs are generally at low risk of developing disease following exposure to anthrax. When disease does occur, it appears associated with oral exposure to the bacteria leading to massive swelling of the head, neck, and mediastinal regions. Death is due to toxemia and shock. For animals at high risk, such as search and rescue dogs with a known exposure, doxycycline at 5 mg/kg orally once daily for 45 to 60 days is suggested as a prophylactic treatment. Additional information on the diagnosis, prevention, and treatment of the disease in dogs is presented. PMID- 16353909 TI - The joint food and agriculture organization of the United Nations/World Health Organization Expert Committee on Food Additives and its role in the evaluation of the safety of veterinary drug residues in foods. AB - The Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) recommended the evaluation of food additives at the international level through the establishment of an expert committee or committees. These committees evaluated the safety of food additives present as residues resulting from the use of pesticides or veterinary pharmaceuticals. The results of these meetings include international harmonization on acceptable daily intake of these compounds and the maximum residue limit that is permitted to be present within any food of animal or plant origin. The decisions rendered by these committees provide a key element in the elimination of international trade barriers associated with products intended for human consumption. PMID- 16353910 TI - Drug compounding for veterinary patients. AB - Drugs have been compounded for veterinary practice for many years because it has been necessary in the course of routine practice. However, regulations and compliance policy guidelines (CPGs) should be recognized. A new CPG issued in July 2003 listed the current Food and Drug Administration (FDA) limitations on compounding for veterinary medicine. To summarize the guideline: drugs must not be compounded from bulk substances, and the compounding must not constitute manufacture of a new animal drug. Drug compounding on a case-by-case basis is allowed under the CPG. However, veterinarians and pharmacists must be aware of potential incompatibilities and practices that may interfere with the drug's stability, purity, and/or potency. PMID- 16353911 TI - Fish drug analysis--Phish-Pharm: a searchable database of pharmacokinetics data in fish. AB - Information about drug residues and pharmacokinetic parameters in aquatic species is relatively sparse. In addition, it is difficult to rapidly compare data between studies due to differences in experimental conditions, such as water temperatures and salinity. To facilitate the study of aquatic species drug metabolism, we constructed a Fish Drug/Chemical Analysis Phish-Pharm (FDA-PP) database. This database consists of more than 400 articles that include data from 90 species (64 genera) of fish. Data fields include genus, species, water temperatures, the average animal weight, sample types analyzed, drug (or chemical) name, dosage, route of administration, metabolites identified, method of analysis, protein binding, clearance, volume of distribution in a central compartment (Vc) or volume of distribution at steady-state (Vd), and drug half lives (t((1/2))). Additional fields list the citation, authors, title, and Internet links. The document will be periodically updated, and users are invited to submit additional data. Updates will be announced in future issues of The AAPS Journal. This database will be a valuable resource to investigators of drug metabolism in aquatic species as well as government and private organizations involved in the drug approval process for aquatic species. PMID- 16353913 TI - Challenges with the development and approval of pharmaceuticals for fish. AB - With an increase in consumer recognition of the health benefits associated with seafood consumption, the volume of fisheries and aquaculture products consumed by the average American is expected to rise. With a concomitant expectation for high quality products, aquaculture is likely to become a greater source of consumed fish. As the United States aquaculture industry grows, so does the need to provide veterinary services. As with any intensive farming system, appropriate medications are needed to maintain animal health and to manage fish populations. This article introduces some of the challenges associated with drug approvals for aquatic species and describes how the process of development and regulation of drugs for use in aquatic animals differs from that associated with uses in terrestrial species. PMID- 16353912 TI - Use of anatomical and kinetic models in the evaluation of human food additive safety. AB - Toxicological testing in animals is relied upon as a surrogate for clinical testing of most food additives. Both animal and human clinical test results are generally available for direct additives when high levels of exposure are expected. Limited animal studies or in vitro test results may be the only sources of toxicological data available when low levels of exposure (microg/person/day) are expected and where no effects of the additive on the food itself are desired. Safety assessment of such materials for humans requires mathematical extrapolation from any effects observed in test animals to arrive at acceptable daily intakes (ADIs) for humans. Models of anatomy may be used to estimate tissue and organ weights where that information is missing and necessary for evaluation of a data set. The effect of growth on target tissue exposure during critical phases of organ development can be more accurately assessed when models of growth and known physiological changes are combined with pharmacokinetic results for test species. Kinetic models, when combined with limited chemical property, kinetic, and distribution data, can often be used to predict steady-state plasma and tissue levels of a test material over the range of doses employed in chronic studies to aid in interpretation of effects that are often nonlinear with respect to delivered dose. A better understanding of the reasons for nonlinearity of effects in animals improves our confidence in extrapolation to humans. PMID- 16353914 TI - Small-molecule agonists and antagonists of the opioid receptor-like receptor (ORL1, NOP): ligand-based analysis of structural factors influencing intrinsic activity at NOP. AB - The recently discovered fourth member of the opioid receptor family, the nociceptin receptor (NOP) and its endogenous ligand, the heptadecapeptide nociceptin, are involved in several central nervous system pathways, such as nociception, reward, tolerance, and feeding. The discovery of small-molecule ligands for NOP is being actively pursued for several therapeutic applications. This review presents a brief overview of the several recently reported NOP ligands, classified as NOP agonists and antagonists, with an emphasis on the analysis of the structural features that may be important for modulating the agonist/antagonist profile (intrinsic activity) of these ligands. Structure activity relationships in our own series of dihydroindolinone-based NOP ligands and those of the various reported ligands indicate that the lipophilic substituent on the common basic nitrogen present in all NOP ligands plays a role in determining the agonist/antagonist profile of the NOP ligand. This analysis provides a basis for the rational drug design of NOP ligands of desired intrinsic activity and provides a framework for developing pharmacophore models for high affinity binding and intrinsic activity at the NOP receptor. Since NOP agonists and antagonists both have therapeutic value, rational approaches for obtaining both within a high-affinity binding class of compounds are very useful for designing potent and selective NOP ligands with the desired profile of intrinsic efficacy. PMID- 16353915 TI - DARPP-32 mediates the actions of multiple drugs of abuse. AB - Drugs of abuse share the ability to enhance dopaminergic neurotransmission in the dorsal and ventral striatum. The action of dopamine is modulated by additional neurotransmitters, including glutamate, serotonin and adenosine. All these neurotransmitters regulate the phosphorylation state of Dopamine- and cAMP regulated phosphoprotein, Mr 32 kDa (DARPP-32). Phosphorylation at Thr(34) by protein kinase A converts DARPP-32 into a potent inhibitor of the multifunctional serine/threonine protein phosphatase, PP-1. Phosphorylation at Thr(75) by Cdk5 converts DARPP-32 into an inhibitor of protein kinase A. The state of phosphorylation of DARPP-32 at Thr(34) also depends on the phosphorylation state of Ser(97) and Ser(130), which are phosphorylated by CK2 and CK1, respectively. By virtue of regulation of these 4 phosphorylation sites, and through its ability to modulate the activity of PP-1 and protein kinase A, DARPP-32 plays a key role in integrating a variety of biochemical, electrophysiological, and behavioral responses controlled by dopamine and other neurotransmitters. Importantly, there is now a large body of evidence that supports a key role for DARPP-32-dependent signaling in mediating the actions of multiple drugs of abuse including cocaine, amphetamine, nicotine, caffeine, LSD, PCP, ethanol and morphine. PMID- 16353916 TI - Recommended reading in population pharmacokinetic pharmacodynamics. AB - Developing the skills or expertise to create useful population pharmacokinetic pharmacodynamic models can be a daunting task-the level of mathematical and statistical complexity is such that newcomers to the field are frequently overwhelmed. A good place to start in learning the field is to read articles in the literature. However, the number of articles dealing with population pharmacokinetic pharmacodynamics is exponentially increasing on a yearly basis, so choosing which articles to read can be difficult. The purpose of this review is to provide a recommended reading list for newcomers to the field. The list was chosen based on perceived impact of the article in the field, the quality of the article, or to highlight some important detail contained within the article. After reading the articles in the list, it is believed that the reader will have a broad overview of the field and have a sound foundation for more-detailed reading of the literature. PMID- 16353917 TI - On some "disadvantages" of the population approach. AB - In a seminal article on population pharmacokinetic modeling, researchers demonstrated how means and variances of pharmacokinetic parameters for a patient population could be inferred from sparse data collected under conditions of routine patient care. But they also identified 4 potential concerns about their methodology: unobserved confounding variables may bias the inferences; conditions under which data are collected may lead to inaccuracies of reporting or recording; correlations among important predictor variables may reduce statistical efficiency; and costs cannot be controlled by principles of study design. Experiences are reviewed that relate to these potential disadvantages. A method is presented for diagnosing the possible presence of confounding. A model is constructed and applied that captures the influences of data inaccuracies. An example of selecting from among correlated covariates is summarized. Finally, a methodology for optimal study design is reviewed and applied to an example. PMID- 16353918 TI - Population pharmacokinetics/pharmacodynamics of anesthetics. AB - In this article we review how population pharmacokinetic/pharmacodynamic (PD) modeling has evolved in the specialty of anesthesiology, how anesthesiology benefited from the mixed-effects approach, and which features of modeling need careful attention. Key articles from the anesthesiology literature are selected to discuss the modeling of typical anesthesiological PD end points, such as level of consciousness and analgesia, interactions between hypnotics and analgesics, estimation with poor and sometimes rich data sets from populations of various sizes, covariate detection, covariances between random effects, and Bayesian forecasting. PMID- 16353919 TI - Modeling and simulation of adherence: approaches and applications in therapeutics. AB - Partial adherence with a prescribed or randomly assigned dose gives rise to unintended variability in actual drug exposure in clinical practice and during clinical trials. There are tremendous costs associated with incomplete and/or improper drug intake-to both individual patients and society as a whole. Methodology for quantifying the relation between adherence, exposure and drug response is an area of active research. Modeling and statistical approaches have been useful in evaluating the impact of adherence on therapeutics and in addressing the challenges of confounding and measurement error which arise in this context. This paper reviews quantitative approaches to using adherence information in improving therapeutics. It draws heavily on applications in the area of HIV pharmacology. PMID- 16353920 TI - A pragmatic approach to the design of population pharmacokinetic studies. AB - The publication of a seminal article on nonlinear mixed-effect modeling led to a revolution in pharmacokinetics (PKs) with the introduction of the population approach. Since then, interest in obtaining accurate and precise estimates of population PK parameters has led to work on population PK study design that extended previous work on optimal sampling designs for individual PK parameter estimation. The issues and developments in the design of population PK studies are reviewed as a prelude to investigating, via simulation, the performance of 2 approaches (population Fisher information matrix D-optimal design and informative block [profile] randomized [IBR] design) for designing population PK studies. The results of our simulation study indicate that the designs based on the 2 approaches yielded efficient parameter estimates. The designs based on the 2 approaches performed similarly, and in some cases designs based on the IBR approach were slightly better. The ease with which the IBR designs can be generated makes them preferable in drug development, where pragmatism and time are of great consideration. We, therefore, refer to the IBR designs as pragmatic designs. Pragmatic designs that achieve high efficiency in the estimation parameters should be used in the design of population PK studies, and simulation should be used to determine the efficiency of the designs. PMID- 16353921 TI - Serotonin transporters: implications for antidepressant drug development. AB - Due to the complexity of the disease, several hypotheses exist to explain the etiology of depression. The monoamine theory of depression suggests that disruptions in the serotonergic and noradrenergic systems result in depressive symptoms. Therefore, the serotonin transporter (SERT) has become a pharmacological target for treating these symptoms. This review will discuss what is known about the molecular interactions of antidepressants with SERT. The effects of antidepressants on SERT regulation and expression in addition to the receptors that may be involved in mediating these effects will be addressed. Specifically, how changes to SERT expression following chronic antidepressant treatment may contribute to the therapeutic benefits of antidepressants will be discussed. Furthermore, the effects of SERT gene polymorphisms on antidepressant efficacy will be examined. Finally, a brief overview of other hypotheses of depression will be addressed as well as factors that must be considered for future antidepressant development. PMID- 16353922 TI - Homology modeling of opioid receptor-ligand complexes using experimental constraints. AB - Opioid receptors interact with a variety of ligands, including endogenous peptides, opiates, and thousands of synthetic compounds with different structural scaffolds. In the absence of experimental structures of opioid receptors, theoretical modeling remains an important tool for structure-function analysis. The combination of experimental studies and modeling approaches allows development of realistic models of ligand-receptor complexes helpful for elucidation of the molecular determinants of ligand affinity and selectivity and for understanding mechanisms of functional agonism or antagonism. In this review we provide a brief critical assessment of the status of such theoretical modeling and describe some common problems and their possible solutions. Currently, there are no reliable theoretical methods to generate the models in a completely automatic fashion. Models of higher accuracy can be produced if homology modeling, based on the rhodopsin X-ray template, is supplemented by experimental structural constraints appropriate for the active or inactive receptor conformations, together with receptor-specific and ligand-specific interactions. The experimental constraints can be derived from mutagenesis and cross-linking studies, correlative replacements of ligand and receptor groups, and incorporation of metal binding sites between residues of receptors or receptors and ligands. This review focuses on the analysis of similarity and differences of the refined homology models of mu, delta, and kappa-opioid receptors in active and inactive states, emphasizing the molecular details of interaction of the receptors with some representative peptide and nonpeptide ligands, underlying the multiple modes of binding of small opiates, and the differences in binding modes of agonists and antagonists, and of peptides and alkaloids. PMID- 16353923 TI - Neuropeptide-processing enzymes: applications for drug discovery. AB - Neuropeptides serve many important roles in communication between cells and are an attractive target for drug discovery. Neuropeptides are produced from precursor proteins by selective cleavages at specific sites, and are then broken down by further cleavages. In general, the biosynthetic cleavages occur within the cell and the degradative cleavages occur postsecretion, although there are exceptions where intracellular processing leads to inactivation, or extracellular processing leads to activation of a particular neuropeptide. A relatively small number of peptidases are responsible for processing the majority of neuropeptides, both inside and outside of the cell. Thus, inhibition of any one enzyme will lead to a broad effect on several different neuropeptides and this makes it unlikely that such inhibitors would be useful therapeutics. However, studies with mutant animals that lack functional peptide-processing enzymes have facilitated the discovery of novel neuropeptides, many of which may be appropriate targets for therapeutics. PMID- 16353924 TI - Evidence of effectiveness: how much can we extrapolate from existing studies? AB - Drug development can be a science of extrapolation if the use of a drug exposure response relationship is embraced and implemented through mechanistically oriented pharmacokinetic (PK)-pharmacodynamic (PD) modeling analysis and clinical trial simulation. The traditional requirement of at least 2 adequate and well controlled phase III studies by the US Food and Drug Administration for drug approval can be waived in certain situations, substantially reducing the resources and time. In this article, the authors introduce a real drug development case where the chance for this exemption was maximized by actively using PK-PD modeling followed by clinical trial simulation, resulting in faster and more economical introduction of a new dosage regimen to patients. PMID- 16353926 TI - Challenges in the transition to model-based development. AB - Practitioners of the art and science of pharmacometrics are well aware of the considerable effort required to successfully complete modeling and simulation activities for drug development programs. This is particularly true because of the current, ad hoc implementation wherein modeling and simulation activities are piggybacked onto traditional development programs. This effort, coupled with the failure to explicitly design development programs around modeling and simulation, will continue to be an important obstacle to the successful transition to model based drug development. Challenges with timely data availability, high data discard rates, delays in completing modeling and simulation activities, and resistance of development teams to the use of modeling and simulation in decision making are all symptoms of an immature process capability for performing modeling and simulation. A process that will fulfill the promise of model-based development will require the development and deployment of three critical elements. The first is the infrastructure--the data definitions and assembly processes that will allow efficient pooling of data across trials and development programs. The second is the process itself--developing guidelines for deciding when and where modeling and simulation should be applied and the criteria for assessing performance and impact. The third element concerns the organization and culture--the establishment of truly integrated, multidisciplinary, and multiorganizational development teams trained in the use of modeling and simulation in decision-making. Creating these capabilities, infrastructure, and incentivizations are critical to realizing the full value of modeling and simulation in drug development. PMID- 16353925 TI - Population pharmacokinetic studies in pediatrics: issues in design and analysis. AB - The current review addresses the following 3 frequently encountered challenges in the design and analysis of population pharmacokinetic studies in pediatrics: (1) body size adjustments during the development of pharmacostatistical models, (2) design and validation of limited sampling strategies, and (3) the integration of historical priors in data analysis and trial simulation. Size adjustments with empiric approaches based on body weight or body surface area have frequently proven as a pragmatic tool to overcome large size differences in a pediatric study population. Allometric size adjustments, however, provide a more mechanistic, physiologically based approach that, if used a priori, allows delineation of the effect of size from that of other covariates that show a high degree of collinearity. The frequent lack of dense data sets in pediatric clinical pharmacology because of ethical and logistic constraints in study design can be overcome with the application of D-optimality-based limited sampling schemes in combination with Bayesian and nonlinear mixed-effects modeling approaches. Empirically based dose selection and clinical trial designs for pediatric clinical pharmacology studies can be improved by applying clinical trial simulation techniques, especially if they integrate adult and pediatric in vitro and/or in vivo data as historic priors. Although integration of these concepts and techniques in population pharmacokinetic analyses is not only limited to pediatric research, their application allows researchers to overcome some major hurdles frequently encountered in pharmacokinetic studies in pediatrics and, thus, provides the basis for additional clinical pharmacology research in this previously insufficiently studied fraction of the general population. PMID- 16353927 TI - Further advances in the synthesis of endocannabinoid-related ligands. AB - Recent advances in the synthesis of endocannabinoid-related ligands for the period 2001-2004 are covered in this review. During this period the first solid phase synthesis of anandamide (AEA) analogs was developed, which allows modification at both the head group and the end pentyl chain. Synthesis of water soluble prodrugs of noladin ether was reported, which are chemically stable, rapidly release noladin ether under enzymatic conditions and are shown to reduce intraocular pressure. The structure-activity relationships (SAR) of alkylcarbamic acid aryl esters and the discovery of potent archidonylsulfonyl derivatives as fatty acid amide hydrolase (FAAH) inhibitors are summarized. Recent synthetic developments in the controversial area of anandamide membrane transporter (AMT) inhibitors are also discussed. PMID- 16353928 TI - Impact of pharmacometrics on drug approval and labeling decisions: a survey of 42 new drug applications. AB - The value of quantitative thinking in drug development and regulatory review is increasingly being appreciated. Modeling and simulation of data pertaining to pharmacokinetic, pharmacodynamic, and disease progression is often referred to as the pharmacometrics analyses. The objective of the current report is to assess the role of pharmacometrics at the US Food and Drug Administration (FDA) in making drug approval and labeling decisions. The New Drug Applications (NDAs) submitted between 2000 and 2004 to the Cardio-renal, Oncology, and Neuropharmacology drug products divisions were surveyed. For those NDA reviews that included a pharmacometrics consultation, the clinical pharmacology scientists ranked the impact on the regulatory decision(s). Of about a total of 244 NDAs, 42 included a pharmacometrics component. Review of NDAs involved independent, quantitative evaluation by FDA pharmacometricians, even when such analysis was not conducted by the sponsor. Pharmacometric analyses were pivotal in regulatory decision making in more than half of the 42 NDAs. Of the 14 reviews that were pivotal to approval related decisions, 5 identified the need for additional trials, whereas 6 reduced the burden of conducting additional trials. Collaboration among the FDA clinical pharmacology, medical, and statistical reviewers and effective communication with the sponsors was critical for the impact to occur. The survey and the case studies emphasize the need for early interaction between the FDA and sponsors to plan the development more efficiently by appreciating the regulatory expectations better. PMID- 16353929 TI - Model-based development of gemcabene, a new lipid-altering agent. AB - The purpose of this study was to evaluate the value of model-based, quantitative decision making during the development of gemcabene, a novel lipid-altering agent. The decisions were driven by a model of the likely clinical profile of gemcabene in comparison with its competitors, such as 3-hydroxymethylglutaryl coenzyme A reductase inhibitors (statins), the cholesterol absorption inhibitor ezetimibe, and their combination. Dose-response models were developed for the lipid effects (low-density lipoprotein cholesterol [LDL-C] and high-density lipoprotein cholesterol); adverse effects, such as persistent alanine aminotransferase elevation and myalgia; tolerability issues, such as headache; and risk reduction for coronary artery disease-related events for 5 statins, ezetimibe, gemcabene, and their combinations. The integrated model was based on the joint analysis of publicly available summary-level data and proprietary patient-level data and included information from almost 10,000 patients. The model was made available and accessible to the development team by using the Pharsight Drug Model Explorer model visualization technology. The modeling greatly enhanced the understanding of the clinical profile of gemcabene when given alone or in combination with a statin. The interaction between statins and gemcabene for the LDL-C lowering effect was found to be significantly different from the interaction between statins and ezetimibe. Ezetimibe was found to have a pharmacological-independent interaction resulting in additional LDL-C lowering over the entire statin dose range. The gemcabene interaction was found to be less than independent, resulting in almost no additional LDL-C lowering at high-statin doses, although the drug has a significant LDL-C effect when administered alone or in combination with a low dose of a statin. The quick availability of the model after completion of the first phase II trial in the target patient population and the ability of the team to explore the potential clinical efficacy and safety of gemcabene in comparison with alternative treatment options facilitated a quick decision to stop development. PMID- 16353930 TI - A new equivalence based metric for predictive check to qualify mixed-effects models. AB - The main objective of any modeling exercise is to provide a rationale for effective decision making during drug development. The aim of the current simulation experiment was to evaluate the properties of predictive check as a covariate model qualification technique and, more importantly, to introduce and evaluate alternative criteria to qualify models.Original concentration-time profiles (yod) were simulated using a 1-compartment model for an intravenous drug administered to 25 men and 25 women. The typical clearance for male subjects (TVCLm) was assumed to be 5-fold higher than that for female subjects (TVCLf). Fifty such trials under the same design were generated randomly. Predictive check was used as the model qualification tool to study predictive performance of true (males not equal females) and false (males = females) models in the context of maximum likelihood estimation. For each yod, 200 replications were generated to study the properties of a discrepancy variable, a statistic that depends on the model, and a test statistic, a statistic that does not depend on the model. Several qualification criteria were evaluated in assessing predictive performance, such as, predictive p-value (Pp), probability of equivalence (peqv), and probability of rejecting the null hypothesis (data = model) using the Kolmogorov-Smirnov test (pks). The Pp value was calculated using sum of squared errors as a discrepancy variable. For both of the models, the Pp values uniformly ranged between 0 and 1. The pattern of Pp values suggests that qualification of the false model is unlikely. For both of the models, the range of peqv is about 0.95 to 1.0 for concentration at 0.5 hours. However, this is not the case for the concentration at 4 hours, which is primarily dependent on the clearance. The false model (0.35 to 0.50) has poor predictive performance compared with the true model (0.65 to 0.80) using peqv. The pks suggests no difference in the distributions of replicated and original concentrations at all of the time points for both of the models. Discrepancy variables cannot aid in rejecting false models, whereas the use of a test statistic can aid in rejecting false models. However, selection of an informative test statistic is challenging. As far as the qualification criteria are considered, the equivalence-based comparison of a test statistic is more informative than a significance-based comparison. No convincing evidence exists in the literature demonstrating the added advantages of predictive check as a routine model qualification tool over the existing tools, such as diagnostic plots or mechanistic reasoning. However, when a model is to be used for designing a trial, it should at least be able to regenerate the data used to build the model. In such cases, predictive check might offer insights into potential inconsistencies. PMID- 16353932 TI - Pharmacokinetics/Pharmacodynamics and the stages of drug development: role of modeling and simulation. AB - Pharmacokinetic (PK) and pharmacodynamic (PD) modeling and simulation (M&S) are well-recognized powerful tools that enable effective implementation of the learn and-confirm paradigm in drug development. The impact of PK/PD M&S on decision making and drug development risk management is dependent on the question being asked and on the availability and quality of data accessible at a particular stage of drug development. For instance, M&S methodologies can be used to capture uncertainty and use the expected variability in PK/PD data generated in preclinical species for projection of the plausible range of clinical dose; clinical trial simulation can be used to forecast the probability of achieving a target response in patients based on information obtained in early phases of development. Framing the right question and capturing the key assumptions are critical components of the "learn-and-confirm" paradigm in the drug development process and are essential to delivering high-value PK/PD M&S results. Selected works of PK/PD modeling and simulation from preclinical to phase III are presented as case examples in this article. PMID- 16353931 TI - Toward the prediction of CNS drug-effect profiles in physiological and pathological conditions using microdialysis and mechanism-based pharmacokinetic pharmacodynamic modeling. AB - Our ultimate goal is to develop mechanism-based pharmacokinetic (PK) pharmacodynamic (PD) models to characterize and to predict CNS drug responses in both physiologic and pathologic conditions. To this end, it is essential to have information on the biophase pharmacokinetics, because these may significantly differ from plasma pharmacokinetics. It is anticipated that biophase kinetics of CNS drugs are strongly influenced by transport across the blood-brain barrier (BBB). The special role of microdialysis in PK/PD modeling of CNS drugs lies in the fact that it enables the determination of free-drug concentrations as a function of time in plasma and in extracellular fluid of the brain, thereby providing important data to determine BBB transport characteristics of drugs. Also, the concentrations of (potential) extracellular biomarkers of drug effects or disease can be monitored with this technique. Here we describe our studies including microdialysis on the following: (1) the evaluation of the free drug hypothesis; (2) the role of BBB transport on the central effects of opioids; (3) changes in BBB transport and biophase equilibration of anti-epileptic drugs; and (4) the relation among neurodegeneration, BBB transport, and drug effects in Parkinson's disease progression. PMID- 16353933 TI - Opioid peptide-derived analgesics. AB - Two recent developments of opioid peptide-based analgesics are reviewed. The first part of the review discusses the dermorphin-derived, cationic-aromatic tetrapeptide H-Dmt-D-Arg-Phe-Lys-NH(2) ([Dmt(1)]DALDA, where Dmt indicates 2',6' dimethyltyrosine), which showed subnanomolar mu receptor binding affinity, extraordinary mu receptor selectivity, and high mu agonist potency in vitro. In vivo, [Dmt(1)]DALDA looked promising as a spinal analgesic because of its extraordinary antinociceptive effect (3000 times more potent than morphine) in the mouse tail-flick assay, long duration of action (4 times longer than morphine), and lack of effect on respiration. Unexpectedly, [Dmt(1)]DALDA also turned out to be a potent and long-acting analgesic in the tail-flick test when given subcutaneously (s.c.), indicating that it is capable of crossing the blood brain barrier. Furthermore, little or no cross-tolerance was observed with s.c. [Dmt(1)]DALDA in morphine-tolerant mice. The second part of the review concerns the development of mixed mu agonist/delta antagonists that, on the basis of much evidence, are expected to be analgesics with a low propensity to produce tolerance and physical dependence. The prototype pseudopeptide H-Dmt TicPsi[CH(2)NH]Phe-Phe-NH(2) (DIPP-NH(2)[Psi], where Tic indicates 1,2,3,4 tetrahydroisoquinoline-3-carboxylic acid) showed subnanomolar mu and delta receptor binding affinities and the desired mu agonist/delta antagonist profile in vitro. DIPP-NH(2)[Psi] produced a potent analgesic effect after intracerebroventricular administration in the rat tail-flick assay, no physical dependence, and less tolerance than morphine. The results obtained with DIPP NH(2)[Psi] indicate that mixed mu agonist/delta antagonists look promising as analgesic drug candidates, but compounds with this profile that are systemically active still need to be developed. PMID- 16353934 TI - Mucosal delivery of bacterial antigens and CpG oligonucleotides formulated in biphasic lipid vesicles in pigs. AB - The ineffectiveness of simple delivery of soluble antigens to mucosal membranes for immunization has stimulated extensive studies of strategies for appropriate delivery systems and adjuvants. Biphasic lipid vesicles are formulations suitable for the delivery of proteins, peptides, and oligo/polynucleotides. The purpose of these studies was to investigate the ability of biphasic lipid vesicles (as vaccine-targeting adjuvants) containing a bacterial antigen and unmethylated oligonucleotides containing CGdinucleotides - CpG motifs (CpG ODNs) to induce systemic and mucosal immune responses in pigs. Results showed that while the protein, either alone or with CpG ODNs, did not induce mucosal immune responses, administration of antigen and CpG ODNs in biphasic lipid vesicles resulted in induction of both systemic and local antibody responses after immunization using a combined mucosal/systemic approach. PMID- 16353935 TI - RNAi-directed inhibition of DC-SIGN by dendritic cells: prospects for HIV-1 therapy. AB - Drug-resistant human immunodeficiency virus (HIV) infections are increasing globally, especially in North America. Therefore, it is logical to develop new therapies directed against HIV binding molecules on susceptible host cells in addition to current treatment modalities against virus functions. Inhibition of the viral genome can be achieved by degrading or silencing posttranslational genes using small interfering (si) ribonucleic acids (RNAs) consisting of double stranded forms of RNA. These siRNAs usually contain 21-23 base pairs (bp) and are highly specific for the nucleotide sequence of the target messenger RNA (mRNA). These siRNAs form a complex with helicase and nuclease enzymes known as "RNA induced silencing complex" (RISC) that leads to target RNA degradation. Thus, siRNA has become a method of selective destruction of HIV now used by various investigators around the globe. However, given the sequence diversity of the HIV genomes of infected subjects, it is difficult to target a specific HIV sequence. Therefore, targeting nonvariable HIV binding receptors on susceptible cells or other molecules of host cells that are directly or indirectly involved in HIV infections may be an interesting alternative to targeting the virus itself. Thus, the simultaneous use of siRNAs specific for HIV and host cells may be a unique, new approach to the therapy of HIV infections. In this article, we present evidence that siRNA directed at the CD4 independent attachment receptor (DC-SIGN) significantly inhibits HIV infection of dendritic cells (DCs). This effect may be mediated by modulation of p38 mitogen activated protein kinase (MAPK). PMID- 16353936 TI - Recent advances for the treatment of cocaine abuse: central nervous system immunopharmacotherapy. AB - Cocaine addiction continues to be a major health and societal problem in spite of governmental efforts devoted toward educating the public of the dangers of illicit drug use. A variety of pharmacotherapies and psychosocial programs have been proposed in an effort to provide a method for alleviation of the physical and psychological symptoms of cocaine abuse. Unfortunately, these methods have been met with limited success, illustrating a critical need for new effective approaches for the treatment of cocaine addiction. Recently an alternative cocaine abuse treatment strategy was proposed using intranasal administration of an engineered filamentous bacteriophage displaying cocaine-sequestering antibodies on its surface. These phage particles are an effective vector for CNS penetration and are capable of binding cocaine, thereby blocking its behavioral effects in a rodent model. The convergence of phage display and immunopharmacotherapy has allowed for an investigation of the efficacy of protein based therapeutics acting within the CNS on the effects of cocaine in animal models and has uncovered a new tool in the battle against cocaine addiction. PMID- 16353937 TI - Mu opioid receptor regulation and opiate responsiveness. AB - Opiate drugs such as morphine are well known for their ability to produce potent analgesia as well as such unwanted side effects as tolerance, physical dependence, respiratory suppression and constipation. Opiates act at opioid receptors, which belong to the family of G protein-coupled receptors. The mechanisms governing mu opioid receptor (muOR) regulation are of particular interest since morphine and other clinically important analgesics produce their pharmacological effects through this receptor. Here we review recent advances in understanding how opioid receptor regulation can impart differential agonist efficacy produced in vivo. PMID- 16353938 TI - Kappa opioids as potential treatments for stimulant dependence. AB - Stimulant abuse is a major problem in the United States and the development of pharmacological treatments for stimulant abuse remains an important therapeutic goal. Classically, the "dopamine hypothesis" has been used to explain the development of addiction and dependence of stimulants. This hypothesis involves the direct increase of dopamine as the major factor in mediating the abuse effects. Therefore, most treatments have focused on directly influencing the dopamine system. Another approach, which has been explored for potential treatments of stimulant abuse, is the use of kappa opioid agonists. The kappa receptor is known to be involved, via indirect effects, in synaptic dopamine levels. This review covers several classes of kappa opioid ligands that have been explored for this purpose. PMID- 16353939 TI - Lung microdialysis--a powerful tool for the determination of exogenous and endogenous compounds in the lower respiratory tract (mini-review). AB - In vivo measurement of concentrations of drugs and endogenous substances at the site of action has become a primary focus of research. In this context the minimal invasive microdialysis (MD) technique has been increasingly employed for the determination of pharmacokinetics in lung. Although lung MD is frequently employed to investigate various drugs and endogenous substances, the majority of lung MD studies were performed to determine the pharmacokinetic profile of antimicrobials that can be related to the importance of respiratory tract infections. For the lower respiratory tract various methods, such as surgical collection of whole lung tissue and bonchoalveolar lavage (BAL), are currently available for the determination of pharmacokinetics of antimicrobials. Head-to head comparison of pharmacokinetics of antibiotics in lung revealed high differences between MD and conventional methods. MD might be regarded as a more advantageous approach because of its higher anatomical resolution and the ability to obtain dynamic time-vs-concentration profiles within one subject. However, due to ethical objections lung MD is limited to animals or patients undergoing elective thoracic surgery. From these studies it was speculated that the concentrations in healthy lung tissue may be predicted reasonably by the measurement of concentrations in skeletal muscle tissue. However, until now this was only demonstrated for beta-lactam antibiotics and needs to be confirmed for other classes of antimicrobials. In conclusion, the present review shows that MD is a promising method for the determination of antimicrobials in the lung, but might also be applicable for measuring a wide range of other drugs and for the investigation of metabolism in the lower respiratory tract. PMID- 16353940 TI - Pharmacokinetic-pharmacodynamic modeling in the data analysis and interpretation of drug-induced QT/QTc prolongation. AB - In this review, factors affecting the QT interval and the methods that are currently in use in the analysis of drug effects on the QT interval duration are overviewed with the emphasis on (population) pharmacokinetic-pharmacodynamic (PK PD) modeling. Among which the heart rate (HR) and the circadian rhythm are most important since they may interfere with the drug effect and need to be taken into account in the data analysis. The HR effect or the RR interval (the distance between 2 consecutive R peaks) effect is commonly eliminated before any further analysis, and many formulae have been suggested to correct QT intervals for changes in RR intervals. The most often used are Bazett and Fridericia formulae introduced in 1920. They are both based on the power function and differ in the exponent parameter. However, both assume the same exponent for different individuals. More recent findings do not confirm this assumption, and individualized correction is necessary to avoid under- or overcorrection that may lead to artificial observations of drug-induced QT interval prolongation. Despite the fact that circadian rhythm in QT and QTc intervals is a well-documented phenomenon, it is usually overlooked when drug effects are evaluated. This may result in a false-positive outcome of the analysis as the QTc peak due to the circadian rhythm may coincide with the peak of the drug plasma concentration. In view of these effects interfering with a potential drug effect on the QTc interval and having in mind low precision of QT interval measurements, a preferable way to evaluate the drug effect is to apply a population PK-PD modeling. In the literature, however, there are only a few publications in which population PK-PD modeling is applied to QT interval prolongation data, and they all refer to antiarrhythmic agents. In this review, after the most important sources of variability are outlined, a comprehensive population PK-PD model is presented that incorporates an individualized QT interval correction, a circadian rhythm in the individually corrected QT intervals, and a drug effect. The model application is illustrated using real data obtained with 2 compounds differing in their QT interval prolongation potential. The usefulness of combining data of several studies is stressed. Finally, the standard approach based on the raw observations and formal statistics, as described in the Preliminary Concept paper of the International Conference on Harmonization, is briefly compared with the method based on population PK-PD modeling, and the advantages of the latter are outlined. PMID- 16353942 TI - In vivo microdialysis for PK and PD studies of anticancer drugs. AB - In vivo microdialysis technique has become one of the major tools to sample endogenous and exogenous substances in extracellular spaces. As a well-validated sampling technique, microdialysis has been frequently employed for quantifying drug disposition at the desired target in both preclinical and clinical settings. This review addresses general methodological considerations critical to performing microdialysis in tumors, highlights selected preclinical and clinical studies that characterized drug disposition in tumors by the use of microdialysis, and illustrates the potential application of microdialysis in the assessment of tumor response to cancer treatment. PMID- 16353943 TI - Anion-induced water flux as drug release mechanism through cationic Eudragit RS 30D film coatings. AB - The objective of this study was to investigate the anion-controlled drug release mechanism through the cationic coating polymer Eudragit RS 30 D as a function of the anion attraction toward the polymer's quarternary ammonium group (QAG), anion valence, and film composition. The mechanism was investigated by dissolution testing, determination of chloride ion exchange using ion chromatography, plasticizer leaching by means of differential scanning calorimetry, and water uptake by Karl Fischer titration. All experiments were performed on coated theophylline micro tablets or isolated films of various compositions using 0.01 M sodium nitrate, sodium sulfate, disodium succinate, sodium acetate, and succinic acid as dissolution media. The mechanism of drug release involved an immediate penetration of dissolution medium into the polymer followed by an instant exchange of chloride against the medium's anion species at completely different rates compared with the drug release. Dependent on the attraction of the anion toward the QAGs, a water flux was induced by back and forth exchanging anions. Strong attraction (nitrate, sulfate) resulted in a low water flux while weak attraction resulted in a high flux (acetate, succinic acid). The water flux increased at increasing number of QAGs. Plasticizer acted as a diluent in respect of the number of QAGs, thus higher plasticizer concentrations led to lower drug release. PMID- 16353944 TI - Molecular modeling of mono- and bis-quaternary ammonium salts as ligands at the alpha4beta2 nicotinic acetylcholine receptor subtype using nonlinear techniques. AB - The neuronal nicotinic acetylcholine receptor (nAChR) has been a target for drug development studies for over a decade. A series of mono- and bis-quaternary ammonium salts, known to be antagonists at nAChRs, were separated into 3 structural classes and evaluated using both self-organizing map (SOM) and genetic functional approximation (GFA) algorithm models. Descriptors from these compounds were used to create several nonlinear quantitative structure-activity relationships (QSARs). The SOM methodology was effective in appropriately grouping these compounds with diverse structures and activities. The GFA models were also able to predict the activities of these molecules. Charge distribution and the hydrophobic free energies were found to be important indicators of bioactivity for this particular class of molecules. These QSAR approaches may be a useful to screen and select in silico new drug candidates from larger compound libraries to be further evaluated in in vitro biological assays. PMID- 16353945 TI - Microdialysis of large molecules. AB - Microdialysis has been used in many tissues, including skin, brain, adipose tissue, muscle, kidney, and gastrointestinal tract, to recover low-molecular mass endogenous mediators, metabolites, and xenobiotics from the interstitial space. Recently, molecules of larger molecular mass, such as plasma proteins, cytokines, growth factors, and neuropeptides, have also been recovered successfully using larger-pore membranes. Microdialysis recovery of large molecules offers the opportunity to identify patterns of protein expression in a variety of tissue spaces and to evaluate clinically useful biomarkers of disease. From this may develop a better understanding of the disease process and its diagnosis and more targeted approaches to therapy. PMID- 16353941 TI - The therapeutic potential of drugs that target cannabinoid receptors or modulate the tissue levels or actions of endocannabinoids. AB - There are at least 2 types of cannabinoid receptor, CB(1) and CB(2), both G protein coupled. CB(1) receptors are expressed predominantly at nerve terminals and mediate inhibition of transmitter release, whereas CB(2) receptors are found mainly on immune cells, their roles including the modulation of cytokine release and of immune cell migration. Endogenous agonists for cannabinoid receptors also exist. These "endocannabinoids" are synthesized on demand and removed from their sites of action by cellular uptake and intracellular enzymic hydrolysis. Endocannabinoids and their receptors together constitute the endocannabinoid system. This review summarizes evidence that there are certain central and peripheral disorders in which increases take place in the release of endocannabinoids onto their receptors and/or in the density or coupling efficiency of these receptors and that this upregulation is protective in some disorders but can have undesirable consequences in others. It also considers therapeutic strategies by which this upregulation might be modulated to clinical advantage. These strategies include the administration of (1) a CB(1) and/or CB(2) receptor agonist or antagonist that does or does not readily cross the blood brain barrier; (2) a CB(1) and/or CB(2) receptor agonist intrathecally or directly to some other site outside the brain; (3) a partial CB(1) and/or CB(2) receptor agonist rather than a full agonist; (4) a CB(1) and/or CB(2) receptor agonist together with a noncannabinoid, for example, morphine or codeine; (5) an inhibitor or activator of endocannabinoid biosynthesis, cellular uptake, or metabolism; (6) an allosteric modulator of the CB(1) receptor; and (7) a CB(2) receptor inverse agonist. PMID- 16353946 TI - Population pharmacokinetics of recombinant factor XIII in cynomolgus monkeys. AB - Hemostasis in humans and other animals is a complex process that controls blood loss after a vascular injury. Factor XIII (FXIII) stabilizes clots primarily by cross-linking fibrin, thus protecting a newly formed clot from fibrinolysis by plasmin. Congenital deficiencies in humans involving FXIII are associated with delayed bleeding and wound healing and severe spontaneous hemorrhaging. These symptoms can be alleviated by intravenous administration of enriched FXIII plasma fractions. Circulating plasma FXIII is found as a heterotetramer that dissociates in the presence of calcium and thrombin into an active dimer and 2 inactive monomers. The recombinant FXIII under investigation is the active dimer alone. A 3-compartment, nonlinear population pharmacokinetic model was implemented in NONMEM V and then used to analyze data from preclinical studies in cynomolgus monkeys. The model simultaneously describes endogenous production of dimer (0.622 microg kg(-1) hr(-1)) and monomer (12.1 microg kg(-1) hr(-1)), and the administration of recombinant dimer. The model incorporates the rate and extent of complexation of recombinant dimer with available endogenous monomer (6.59 mg( 1) kg hr(-1)) to form the heterotetramer. Half-lives for dimer, heterotetramer, and monomer (3.33 hours, 2.83 days, and 3.94 hours for A(2), A(2)B(2), and B, respectively) were estimated, along with their variability in the population studied. PMID- 16353948 TI - Patent issues in drug development: perspectives of a pharmaceutical scientist attorney. AB - The major purpose of this article is to emphasize the need for pharmaceutical scientists to have a better understanding of patent fundamentals. This need is illustrated by analyses of key scientific and legal issues that arose during recent patent infringement cases involving Prozac, Prilosec, and Buspar. Economic incentives for drug discovery and development clash with societal needs for low cost pharmaceuticals in the United States and all over the world. The Hatch Waxman Act of 1984 was enacted to promote public health by balancing the interests of brand name and generic companies. Patent protection, which provides a monopoly for a limited time, is aimed to provide such incentives. Creation of patents requires the interaction between scientists and lawyers, an endeavor made difficult by the differing intellectual spheres of their respective disciplines. Therefore, in the first place, a thorough understanding of patent fundamentals among pharmaceutical scientists will help them work more efficiently with patent attorneys. Second, it will enable them to appreciate the strengths and weaknesses of individual patents, which is critical in developing strategies amidst the ongoing patent tug-of-war between brand-name and generic companies. PMID- 16353947 TI - Kappa opioid antagonists: past successes and future prospects. AB - Antagonists of the kappa opioid receptor were initially investigated as pharmacological tools that would reverse the effects of kappa opioid receptor agonists. In the years following the discovery of the first selective kappa opioid antagonists, much information about their chemistry and pharmacology has been elicited and their potential therapeutic uses have been investigated. The review presents the current chemistry, ligand-based structure activity relationships, and pharmacology of the known nonpeptidic selective kappa opioid receptor antagonists. This manuscript endeavors to provide the reader with a useful reference of the investigations made to define the structure-activity relationships and pharmacology of selective kappa opioid receptor antagonists and their potential uses as pharmacological tools and as therapeutic agents in the treatment of disease states. PMID- 16353949 TI - Regulation of monoamine transporters: influence of psychostimulants and therapeutic antidepressants. AB - Synaptic neurotransmission in the central nervous system (CNS) requires the precise control of the duration and the magnitude of neurotransmitter action at specific molecular targets. At the molecular level, neurotransmitter signaling is dynamically regulated by a diverse set of macromolecules including biosynthetic enzymes, secretory proteins, ion channels, pre- and postsynaptic receptors and transporters. Monoamines, 5-hydroxytryptamine or serotonin (5-HT), norepinephrine (NE), and dopamine (DA) play an important modulatory role in the CNS and are involved in numerous physiological functions and pathological conditions. Presynaptic plasma membrane transporters for 5-HT (SERT), NE (NET), and DA (DAT), respectively, control synaptic actions of these monoamines by rapidly clearing the released amine. Monoamine transporters are the sites of action for widely used antidepressants and are high affinity molecular targets for drugs of abuse including cocaine, amphetamine, and 3,4-methylenedioxymetamphetamine (MDMA) "Ecstasy." Monoamine transporters also serve as molecular gateways for neurotoxins. Emerging evidence indicates that regulation of transporter function and surface expression can be rapidly modulated by "intrinsic" transporter activity itself, and antidepressant and psychostimulant drugs that block monoamine transport have a profound effect on transporter regulation. Therefore, disregulations in the functioning of monoamine transporters may underlie many disorders of transmitter imbalance such as depression, attention deficit hyperactivity disorder, and schizophrenia. This review integrates recent progress in understanding the molecular mechanisms of monoamine transporter regulation, in particular, posttranscriptional regulation by phosphorylation and trafficking linked to cellular protein kinases, protein phosphatases, and transporter interacting proteins. The review also discusses the possible role of psychostimulants and antidepressants in influencing monoamine transport regulation. PMID- 16353950 TI - Recognition of psychostimulants, antidepressants, and other inhibitors of synaptic neurotransmitter uptake by the plasma membrane monoamine transporters. AB - The plasma membrane monoamine transporters terminate neurotransmission by removing dopamine, norepinephrine, or serotonin from the synaptic cleft between neurons. Specific inhibitors for these transporters, including the abused psychostimulants cocaine and amphetamine and the tricyclic and SSRI classes of antidepressants, exert their physiological effects by interfering with synaptic uptake and thus prolonging the actions of the monoamine. Pharmacological, biochemical, and immunological characterization of the many site-directed, chimeric, and deletion mutants generated for the plasma membrane monoamine transporters have revealed much about the commonalities and dissimilarities between transporter substrate, ion, and inhibitor binding sites. Mutations that alter the binding affinity or substrate uptake inhibition potency of inhibitors by at least 3-fold are the focus of this review. These findings are clarifying the picture regarding substrate uptake inhibitor/transporter protein interactions at the level of the drug pharmacophore and the amino acid residue, information necessary for rational design of novel medications for substance abuse and a variety of psychiatric disorders. PMID- 16353952 TI - Oral formulation of a novel antiviral agent, PG301029, in a mixture of gelucire 44/14 and DMA (2:1, wt/wt). AB - To develop an oral formulation for PG301029, a novel potent agent for the treatment of Hepatitis C virus infection, that not only has very low aqueous solubility but also degrades rapidly in water. The solubility of PG301029 was determined in water, various aqueous media, and several neat organic solvents. The stability of PG301029 was monitored at room temperature in buffers for 4 days, and in several neat organic solvents for up to 8 mo. Drug concentrations were measured by high-performance liquid chromatography (HPLC). Based on solubility and stability data, Gelucire 44/14 and DMA (N,N-dimethylacetamide) at a weight ratio of 2 to 1 were chosen as the formulation vehicle. After the vehicle was prepared, it was maintained in liquid form at approximately 40 degrees C until the PG301029 was dissolved. The final formulation product was a semisolid at room temperature. The bioavailability of the formulation was tested on 4 female BALB/c mice. PG301029 is insoluble in all tested aqueous media, while its solubility is promising in DMA. This compound is unstable in aqueous media and some organic solvents; however, it is stable in DMA. This proposed formulation is able to hold up to 10 mg/mL of drug and is stable at 4 degrees C. The shelf life for this formulation stored at 4degreesC is extrapolated to be greater than 4 years. This formulation dramatically increases the bioavailability of PG301029. This nonaqueous formulation solves the stability, solubility, and bioavailability problems for PG301029. This semisolid formulation can easily be incorporated into soft elastic capsules. PMID- 16353951 TI - A general procedure for the enantioselective synthesis of the minor tobacco alkaloids nornicotine, anabasine, and anatabine. AB - The minor tobacco alkaloids nornicotine, anabasine, and anatabine from Nicotiana tobacum are known to possess nicotinic receptor agonist activity, although they are relatively less potent than S-(-)-nicotine, the principal tobacco alkaloid. Previous pharmacological investigations and structure-activity studies have been limited owing to the lack of availability of the optically pure forms of these minor alkaloids. We now report a 2-step synthetic procedure for the enantioselective synthesis of the optical isomers of nornicotine and anabasine, and a modified procedure for the synthesis of anatabine enantiomers. These procedures involve initial formation of the chiral ketimine resulting from the condensation of either 1R, 2R, 5R-(+)- or 1S, 2S, 5S-(-)-2-hydroxy-3-pinanone with 3-(aminomethyl)pyridine followed by enantioselective C-alkylation with an appropriate halogenoalkane or halogenoalkene species, N-deprotection, and base catalyzed intramolecular ring closure, to form the appropriate, chirally pure minor tobacco alkaloid. Using this approach, the R-(+)- and S-(-)-enantiomers of the above minor tobacco alkaloids were obtained in good overall chemical yield and excellent enantomeric excess. PMID- 16353953 TI - Eudragit S100 entrapped insulin microspheres for oral delivery. AB - The purpose of this research was to investigate whether Eudragit S100 microspheres have the potential to serve as an oral carrier for peptide drugs like insulin. Microspheres were prepared using water-in oil-in water emulsion solvent evaporation technique with polysorbate 20 as a dispersing agent in the internal aqueous phase and polyvinyl alcohol (PVA)/polyvinyl pyrrolidone as a stabilizer in the external aqueous phase. The use of smaller internal aqueous phase volume (50 microL) and external aqueous-phase volume (25 mL) containing PVA in the manufacturing process resulted in maximum encapsulation efficiency (81.8% +/- 0.9%). PVA-stabilized microspheres having maximum drug encapsulation released 2.5% insulin at pH 1.0 in 2 hours. In phosphate buffer (pH 7.4), microspheres showed an initial burst release of 22% in 1 hour with an additional 28% release in the next 5 hours. The smaller the volumes of internal and external aqueous phase, the lower the initial burst release. The release of drug from microspheres followed Higuchi kinetics. Scanning electron microscopy of PVA-stabilized microspheres demonstrated spherical particles with smooth surface, and laser diffractometry revealed a mean particle size of 32.51 +/- 20 microm. Oral administration of PVA stabilized microspheres in normal albino rabbits (equivalent to 6.6 IU insulin/kg of animal weight) demonstrated a 24% reduction in blood glucose level, with maximum plasma glucose reduction of 76 +/- 3.0% in 2 hours and effect continuing up to 6 hours. The area under the percentage glucose reduction-time curve was 93.75%. Thus, our results indicate that Eudragit S100 microspheres on oral administration can protect insulin from proteolytic degradation in the gastrointestinal tract and produce hypoglycemic effect. PMID- 16353954 TI - Spray-dried poly(D,L-lactide) microspheres containing carboplatin for veterinary use: in vitro and in vivo studies. AB - The aim of this study was the development of a veterinary dosage form constituted by injectable biodegradable microspheres designed for the subcutaneous release of carboplatin, a chemotherapeutic drug. Poly(D,L-lactide) (PDLLA) microspheres were prepared by an emulsification/spray-drying method, using the drug-to-polymer weight ratios 1:9 and 1:5; blank microspheres (1% w/v) were prepared as a comparison. Microparticles were characterized in terms of morphology, encapsulation efficiency, and in vitro drug release behavior. In vivo tests were conducted on rats by subcutaneous injection of microsphere aqueous suspensions. Levels of carboplatin were evaluated both in the skin and in serum. The microparticles obtained had a spherical shape; particle size ranged from 5 to 7 microm, dependent on drug loading. Microspheres were able to control the in vitro release of the drug: approximately 90% to 100% of the carboplatin was released over 30 days. In vivo results showed that the microspheres were able to release high drug amounts locally, and sustained serum levels of drug were also achieved. Based on these results, carboplatin-loaded PDLLA microspheres may be useful for local delivery of the antineoplastic drug to the tumor, avoiding tumor recurrence in small animals, and may decrease the formation of distant metastases. PMID- 16353955 TI - Structure determination and characterization of carbendazim hydrochloride dihydrate. AB - The objective of this study was to synthesize and characterize the hydrochloride salt of carbendazim with the aim of improving the intrinsic solubility of the parent compound. Carbendazim hydrochloride dihydrate was synthesized for the purpose of increasing the aqueous solubility of the parent drug, carbendazim. This was done with the commonly used saturation and cooling method. The structure was determined by single crystal radiograph crystallography, and the hydrochloride salt was found to be a dihydrate. The salt crystallized in a P 2(1) 2(1) 2(1) (#19) space group, which is typical for nonplanar, achiral, and noncentrosymmetric molecules. The asymmetric unit is comprised of 1 molecule each of carbendazim and chloride and 2 water molecules. The carbendazim molecules arrange themselves in a helical structure, with the waters and the chloride molecules in the channel linking the helix. The crystal lattice is held together by numerous hydrogen bonds, as well as van der Waals interactions. The melting point of the salt is 125.6 degrees C. The solubility of the salt is 6.08 mg/mL, which is a thousand-fold increase from the intrinsic solubility (6.11 microg/mL) of the free base. PMID- 16353956 TI - The influence of swelling capacity of superdisintegrants in different pH media on the dissolution of hydrochlorothiazide from directly compressed tablets. AB - The purpose of this study was to investigate the efficiency of superdisintegrants in promoting tablet disintegration and drug dissolution under varied media pH. Significant reductions in the rate and extent of water uptake and swelling were observed for both sodium starch glycolate (Primojel) and croscarmellose sodium (Ac-Di-Sol) in an acidic medium (0.1 N HCl) but not for crospovidone NF (Polyplasdone XL10), a nonionic polymer. When Primojel and Ac-Di-Sol were incorporated in model formulations, a significant increase in tablet disintegration time was observed for slowly disintegrating tablets (lactose-based tablets) but not for the rapidly disintegrating tablets (dicalcium phosphate based tablets). The dissolution rate of the model drug, hydrochlorothiazide, was found highly dependent on both tablet disintegration efficiency and the solubility of base material(s) in the testing medium. A laser diffraction particle size analyzer proved to be an effective tool for determining the intrinsic swelling of disintegrant particles in different media. Water uptake and swelling were confirmed as 2 important functions of superdisintegrants. The reduced water uptake and swelling capacity of disintegrants containing ionizable substituents in an acidic medium can potentially jeopardize their efficiency in promoting tablet disintegration and the drug dissolution rate. PMID- 16353957 TI - Quantitative analysis of film coating in a pan coater based on in-line sensor measurements. AB - A method was developed that enables in-line analysis of film coating thickness on tablets during a pan coating operation. Real-time measurements were made using a diffuse-reflectance near-infrared (NIR) probe positioned inside the pan during the coating operation. Real-time spectra of replicate batches were used for modeling film growth. Univariate analysis provided a simple method for in-line monitoring of the coating process using NIR data. An empirical geometric 2-vector volumetric growth model was developed, which accounts for differential growth on the face and band regions of biconvex tablets. The thickness of the film coat was determined by monitoring the decrease of absorption bands characteristic of a component of the tablet core and monitoring the increase of bands characteristic of a component in the coating material. There was good correlation between values estimated from the NIR data and the measured tablet volumetric growth. In-line measurements allow the coating process to be stopped when a predetermined tablet coating thickness is achieved. PMID- 16353958 TI - In vitro and in vivo evaluation of guar gum matrix tablets for oral controlled release of water-soluble diltiazem hydrochloride. AB - The objective of the study was to develop guar gum matrix tablets for oral controlled release of water-soluble diltiazem hydrochloride. Matrix tablets of diltiazem hydrochloride, using various viscosity grades of guar gum in 2 proportions, were prepared by wet granulation method and subjected to in vitro drug release studies. Diltiazem hydrochloride matrix tablets containing either 30% wt/wt low-viscosity (LM1), 40% wt/wt medium-viscosity (MM2), or 50% wt/wt high-viscosity (HM2) guar gum showed controlled release. The drug release from all guar gum matrix tablets followed first-order kinetics via Fickian-diffusion. Further, the results of in vitro drug release studies in simulated gastrointestinal and colonic fluids showed that HM2 tablets provided controlled release comparable with marketed sustained release diltiazem hydrochloride tablets (D-SR tablets). Guar gum matrix tablets HM2 showed no change in physical appearance, drug content, or in dissolution pattern after storage at 40 degrees C/relative humidity 75% for 6 months. When subjected to in vivo pharmacokinetic evaluation in healthy volunteers, the HM2 tablets provided a slow and prolonged drug release when compared with D-SR tablets. Based on the results of in vitro and in vivo studies it was concluded that that guar gum matrix tablets provided oral controlled release of water-soluble diltiazem hydrochloride. PMID- 16353959 TI - Preparation and characterization of chitosan and trimethyl-chitosan-modified poly (epsilon-caprolactone) nanoparticles as DNA carriers. AB - The purpose of this research was to prepare poly-(epsilon-caprolactone) (PCL) particles by an emulsion-diffusion-evaporation method using a blend of poly (vinyl alcohol) and chitosan derivatives as stabilizers. The chitosan derivatives used were chitosan hydrochloride and trimethyl chitosans (TMC) with varying degrees of quaternization. Particle characteristics-size, zeta potential, surface morphology, cytotoxicity, and transfection efficiency-were investigated. The developed method yields PCL nanoparticles in the size range of 250 to 300 nm with a positive surface charge (2.5 to 6.8 mV). The cytotoxicity was found to be moderate and virtually independent of the stabilizers' concentration with the exception of the highly quaternized TMC (degree of substitution 66%) being significantly more toxic. In immobilization experiments with gel electrophoresis, it could be shown that these cationic nanoparticles (NP) form stable complexes with DNA at a NP:DNA ratio of 3:1. These nanoplexes showed a significantly higher transfection efficiency on COS-1 cells than naked DNA. PMID- 16353960 TI - Effect of humidity on the disintegrant property of alpha-cellulose, part II: a technical note. PMID- 16353961 TI - Design and development of multivesicular liposomal depot delivery system for controlled systemic delivery of acyclovir sodium. AB - The aim of the present study was to design a depot delivery system of acyclovir sodium using multivesicular liposomes (MVLs) to overcome the limitations of conventional therapies and to investigate its in vivo effectiveness for sustained delivery. MVLs of acyclovir were prepared by the reverse phase evaporation method. The loading efficiency of the MVLs (45%-82%) was found to be 3 to 6 times higher than conventional multilamellar vesicles (MLVs). The in vitro release of acyclovir from MVL formulations was found to be in a sustained manner and only 70% of drug was released in 96 hours, whereas conventional MLVs released 80% of drug in 16 hours. Following intradermal administration to Wistar rats, the MVL formulations showed effective plasma concentration for 48 hours compared with MLVs and free drug solution (12-16 hours). C(max) values of MVL formulations were significantly less (8.6-11.4 microg/mL) than MLV and free drug solution (12.5 microg/mL). The AUC(0-48) of the MVL formulations was 1.5- and 3-fold higher compared with conventional liposomes and free drug solution, respectively. Overall, formulations containing phosphatidyl glycerol as negatively charged lipid showed better results. The MVL delivery system as an intradermal depot offers the advantage of a very high loading and controlled release of acyclovir for an extended period of time. The increase in AUC and decrease in C(max) reflects that the MVL formulations could reduce the toxic complications and limitations of conventional iv and oral therapies. PMID- 16353962 TI - The protective effect of lactose on lyophilization of CNK-20402. AB - The goal of this research was to assess the feasibility of using lyophilization to stabilize an exploratory compound, CNK-20402, with a minimal amount of impurity (CNK-20193) formation. A mixed-level full factorial experimental design was used to screen excipients of glycine, mannitol, lactose monohydrate, and povidone K-12. Cryostage microscopy, powder x-ray diffraction, Karl Fischer titration, HPLC, and water vapor sorption were used to assess the formulations' physicochemical properties and stability. Initial physical characterization from powder x-ray diffraction revealed that the mannitol- and glycine-containing formulations were crystalline with the patterns of the pure excipient, whereas the remaining formulations were amorphous in structure. Chemically, the formulations stored at 50 degrees C for 1 month had 2.36%, 1.05%, 0.81%, 0.79%, and 0.49% CNK-20193 for glycine, mannitol, drug alone, povidone K-12, and lactose formulations, respectively. The formulations containing drug-mannitol, drug alone, and drug-lactose were selected for accelerated stability study based on statistical analysis. Recovery of CNK-20193 in these formulations was 1.22%, 1.00%, and 0.55%, respectively, when stored at 40 degrees C/75% relative humidity storage conditions for 3 months. Water vapor sorption analysis revealed weight gains of over 7%, 21%, and 24% for the mannitol, lactose, and drug alone formulations, respectively. Testing formulations with different concentrations of lactose by water vapor sorption indicated that CNK-20402 concentrations as low as 10% (wt/wt) could inhibit the recrystallization of lactose. The lactose containing formulation exhibited the best stability among the formulations tested. The protective mechanism of lactose on the CNK-20402, based on water vapor sorption studies, is believed to be a result of (1) the drug-lactose interaction, and (2) competition between lactose and drug for the residual water in the formulation. PMID- 16353963 TI - Formulation and evaluation of mucoadhesive glipizide microspheres. AB - The purpose of this research was to formulate and systematically evaluate in vitro and in vivo performances of mucoadhesive microspheres of glipizide. Glipizide microspheres containing chitosan were prepared by simple emulsification phase separation technique using glutaraldehyde as a cross-linking agent. Results of preliminary trials indicate that volume of cross-linking agent, time for cross linking, polymer-to-drug ratio, and speed of rotation affected characteristics of microspheres. Microspheres were discrete, spherical, and free flowing. The microspheres exhibited good mucoadhesive property in the in vitro wash-off test and also showed a high percentage drug entrapment efficiency. A 3(2) full factorial design was employed to study the effect of independent variables, polymer-to-drug ratio (X(1) ), and stirring speed (X(2) ) on dependent variables percentage mucoadhesion, t(80), drug entrapment efficiency, and swelling index. The best batch exhibited a high drug entrapment efficiency of 75% and a swelling index of 1.42; percentage mucoadhesion after 1 hour was 78%. The drug release was also sustained for more than 12 hours. The polymer-to-drug ratio had a more significant effect on the dependent variables. In vivo testing of the mucoadhesive microspheres to albino Wistar rats demonstrated significant hypoglycemic effect of glipizide. PMID- 16353964 TI - Conceival, a novel noncontraceptive vaginal vehicle for lipophilic microbicides. AB - The objective of this study was to develop a nontoxic and noncontraceptive vaginal drug delivery vehicle for lipophilic anti-human immunodeficiency virus (HIV) microbicides. Three representative poorly water-soluble novel broad spectrum anti-HIV microbicides, PHI-113, PHI-346, and PHI-443, were evaluated in 11 different solvent systems. Based on their solubility profiles, a novel nonspermicidal self-emulsifying gel (viz Conceival) composed of pharmaceutical excipients, sorbitol, polyethylene glycol 400, polysorbate 80, microcrystalline cellulose, xanthan gum, and water was optimized. Conceival enhanced the solubility of these poorly water-soluble (<0.001 mg/mL) anti-HIV drugs by at least 150- to 270-fold. Conceival was evaluated in vivo in the New Zealand white rabbit model for the preservation of sperm function based on pregnancy outcome and the potential for vaginal irritation following single and multiple intravaginal applications, respectively. Conceival administered intravaginally immediately prior to artificial insemination with semen had no adverse effects on subsequent reproductive performance, neonatal survival, or pup development when compared with untreated control group. Histologic evaluation of vaginal tissues of rabbits exposed intravaginally to Conceival for 14 consecutive days revealed lack of epithelial, submucosal, and vascular changes at the gel application site (total irritation score <3 out of a possible 16). These findings indicate that Conceival has potential to become a clinically useful, safe noncontraceptive vaginal vehicle for lipophilic microbicides. PMID- 16353965 TI - Albumin microspheres as carriers for the antiarthritic drug celecoxib. AB - The present study investigates the preparation of celecoxib-loaded albumin microspheres and the biodistribution of technetium-99m ((99m)Tc)-labeled celecoxib as well as its microspheres after intravenous administration. Microspheres were prepared using a natural polymer BSA using emulsification chemical cross-linking method. The prepared microspheres were characterized for entrapment efficiency, particle size, and in vitro drug release. Surface morphology was studied by scanning electron microscopy. Biodistribution studies were performed by radiolabeling celecoxib (CS) and its microspheres (CMS) using (99m)Tc and injecting arthritic rats intravenously. The geometric mean diameter of the microspheres was found to be 5.46 microm. In vitro release studies indicated that the microspheres sustained the release of the drug for 6 days. Radioactivity measured in different organs after intravenous administration of celecoxib solution showed a significant amount of radioactivity in the liver and spleen. In case of celecoxib-loaded microspheres, a significant amount of radioactivity accumulated in the lungs. No significant difference (P > .1) in the radioactivity was observed between the inflamed joint and the noninflamed joint following intravenous injection of (99m)Tc-CS. However, in case of the microspheres (CMS), the radioactivity present in the inflamed joint was 2.5-fold higher than in the noninflamed joint. The blood kinetic studies revealed that celecoxib-loaded albumin microspheres exhibited prolonged circulation than the celecoxib solution. PMID- 16353966 TI - Licorice: a possible anti-inflammatory and anti-ulcer drug. AB - The purpose of this investigation was to study the anti-inflammatory activities of both glycerrhitinic acid (GA) and the aqueous licorice extract (ALE) in comparison with diclofenac sodium (DS) (10 mg/kg), using the carrageenan-induced paw edema model in male albino rats. In addition, the anti-ulcer activities of ALE, famotidine (FT), and a combination of ALE and FT using indomethacin-induced ulceration technique in rat stomach were investigated. Conventional DS tablets containing GA, as well as DS chewable tablets containing either GA or ALE with different tastes were prepared. Also, rapidly disintegrating FT tablets were prepared using direct compression and camphor sublimation methods. ALE or GA produced significant anti-inflammatory activity similar to DS, and when taken concomitantly, there is no possible antagonism. The anti-ulcer activity of licorice was found to be similar to that of FT in indomethacin-induced ulceration technique in rat stomach. Combination therapy of both FT and licorice showed higher anti-ulcer activity than either of them alone. Generally, tablets containing the crosslinked sodium carboxymethyl cellulose (AcDisol) showed more rapidly disintegrating effect than those including Sodium starch glycolate (Primojel). The oral disintegration was very rapid for all the tested formulations. Also, the amount of FT absorbed from the oral cavity was nearly 9 from 10 mg theoretically present in each formula. It could be concluded that both GA and ALE have anti-inflammatory activity comparable with DS. It may be recommended to add ALE to either FT or diclofinac for more effective anti inflammatory or anti-ulcer formulations, respectively. PMID- 16353967 TI - Physicochemical characterization, in vitro dissolution behavior, and pharmacodynamic studies of rofecoxib-cyclodextrin inclusion compounds. preparation and properties of rofecoxib hydroxypropyl beta-cyclodextrin inclusion complex: a technical note. PMID- 16353968 TI - Bioavailability assessment of ketoprofen incorporated in gelled self-emulsifying formulation: a technical note. PMID- 16353969 TI - Assessment of NIR spectroscopy for nondestructive analysis of physical and chemical attributes of sulfamethazine bolus dosage forms. AB - The goal of this study was to assess the utility of near infrared (NIR) spectroscopy for the determination of content uniformity, tablet crushing strength (tablet hardness), and dissolution rate in sulfamethazine veterinary bolus dosage forms. A formulation containing sulfamethazine, corn starch, and magnesium stearate was employed. The formulations were wet granulated with a 10% (wt/vol) starch paste in a high shear granulator and dried at 60 degrees C in a convection tray dryer. The tablets were compressed on a Stokes B2 rotary tablet press running at 30 rpm. Each sample was scanned in reflectance mode in the wavelengths of the NIR region. Principal component analysis (PCA) of the NIR tablet spectra and the neat raw materials indicated that the scores of the first 2 principal components were highly correlated with the chemical and physical attributes. Based on the PCA model, the significant wavelengths for sulfamethazine are 1514, (1660-1694), 2000, 2050, 2150, 2175, 2225, and 2275 nm; for corn starch are 1974, 2100, and 2325 nm; and for magnesium stearate are 2325 and 2375 nm. In addition, the loadings show large negative peaks around the water band regions ( approximately 1420 and 1940 nm), indicating that the partial least squares (PLS) models could be affected by product water content. A simple linear regression model was able to predict content uniformity with a correlation coefficient of 0.986 at 1656 nm; the use of a PLS regression model, with 3 factors, had an r (2) of 0.9496 and a standard error of calibration of 0.0316. The PLS validation set had an r (2) of 0.9662 and a standard error of 0.0354. PLS calibration models, based on tablet absorbance data, could successfully predict tablet crushing strength and dissolution in spite of varying active pharmaceutical ingredient (API) levels. Prediction plots based on these PLS models yielded correlation coefficients of 0.84 and 0.92 on independent validation sets for crushing strength and Q(120) (percentage dissolved in 120 minutes), respectively. PMID- 16353970 TI - An overview of electrical tomographic measurements in pharmaceutical and related application areas. AB - Tomographic measurement techniques offer the opportunity to quantify the degree of homogeneity of particulate suspensions and other multiphase mixtures. Electrical resistance tomography is a relatively simple and inexpensive technique for measuring the distribution of electrical conductivity within multiphase systems. This can provide pertinent information about the physical form, the chemical composition, or the general status of manufacturing. In this contribution, we present recent applications of this technology to processes in pharmaceutical and related application areas. Examples include on-line measurement of solids distribution in stirred tanks and crystallizers, monitoring the performance of an industrial pressure filter, and flow profile and velocity measurements in a physical model of a catalytic reactor. PMID- 16353971 TI - Feasibility of transdermal delivery of fluoxetine. AB - Feasibility of developing a transdermal drug delivery of fluoxetine has been investigated. Permeation studies of fluoxetine across human cadaver skin were carried out using Franz diffusion cells. The receptor phase consisted of pH 7.4 phosphate buffer maintained at 37 degrees C. Permeation enhancement of fluoxetine, either in the salt or base form, was achieved using various enhancers like azone, SR-38, and ethanol. Various O/W microemulsion systems of fluoxetine were developed to study their effect on the skin permeation of fluoxetine. The results indicated that ethanol at 65% vol/vol was able to increase the permeation of fluoxetine the most, while microemulsion systems showed decrease in the permeation of fluoxetine. The permeation of fluoxetine obtained using a 65% vol/vol ethanolic solution was found to be sufficient to deliver the required dose (20-80 mg) from a patch of feasible size. The results seem promising for developing a transdermal drug delivery system of fluoxetine. PMID- 16353972 TI - Freeze-drying of proteins from a sucrose-glycine excipient system: effect of formulation composition on the initial recovery of protein activity. AB - The purpose of this study was to investigate the effect of sucrose-glycine excipient systems on the stability of selected model proteins during lyophilization. Recovery of protein activity after freeze-drying was examined for the model proteins lactate dehydrogenase and glucose 6-phosphate dehydrogenase in a sucrose-glycine-based excipient system in which the formulation composition was systematically varied. In a sucrose-only excipient system, activity recovery of both model proteins is about 80% and is independent of sucrose concentration over a range from 1 to 40 mg/mL. When both sucrose and glycine are used and the ratio of the 2 excipients is varied, however, activity recovery decreases in a pattern that is consistent with the inhibition of activity recovery by glycine crystals, despite the presence of an adequate amount of sucrose to afford protection. Annealing of sucrose-glycine formulations causes a small but significant decrease in activity recovery relative to unannealed controls, whereas no annealing effect is observed with sucrose-only formulations. Addition of 0.01% polysorbate 80 to the formulation resulted in complete recovery of activity, irrespective of the sucrose-glycine ratio or annealing. Addition of the same concentration of polysorbate 80 to the reconstitution medium caused an increase in activity recovery for each formulation, but the overall pattern remained unchanged. The data are consistent with an interfacial model for lyophilization-associated loss of protein activity involving denaturation at a solid/freeze-concentrate interface. PMID- 16353973 TI - Etoposide-loaded nanoparticles made from glyceride lipids: formulation, characterization, in vitro drug release, and stability evaluation. AB - The aim of the study was to prepare etoposide-loaded nanoparticles with glyceride lipids and then characterize and evaluate the in vitro steric stability and drug release characteristics and stability. The nanoparticles were prepared by melt emulsification and homogenization followed by spray drying of nanodispersion. Spray drying created powder nanoparticles with excellent redispersibility and a minimal increase in particle size (20-40 nm). Experimental variables, such as homogenization pressure, number of homogenization cycles, and surfactant concentration, showed a profound influence on the particle size and distribution. Spray drying of Poloxamer 407-stabilized nanodispersion lead to the formation of matrix-like structures surrounding the nanoparticles, resulting in particle growth. The in vitro steric stability test revealed that the lipid nanoparticles stabilized by sodium tauroglycocholate exhibit excellent steric stability compared with Poloxamer 407. All 3 glyceride nanoparticle formulations exhibited sustained release characteristics, and the release pattern followed the Higuchi equation. The spray-dried lipid nanoparticles stored in black polypropylene containers exhibited excellent long-term stability at 25 degrees C and room light conditions. Such stable lipid nanoparticles with in vitro steric stability can be a beneficial delivery system for intravenous administration as long circulating carriers for controlled and targeted drug delivery. PMID- 16353974 TI - The effect of pH and organic ester penetration enhancers on skin permeation kinetics of terbutaline sulfate from pseudolatex-type transdermal delivery systems through mouse and human cadaver skins. AB - The purpose of this research was to prepare a pseudolatex transdermal delivery system for terbutaline sulfate and to evaluate the effect of pH and organic ester penetration enhancers on permeation kinetics of terbutaline sulfate through mice abdominal skin and human cadaver skin. An increase in the permeation flux by increasing pH was observed. The distribution coefficient of terbutaline sulfate between 1-octanol and buffers of different pH values was also pH-dependent. Furthermore, the change of the permeability coefficient with pH correlated well with the distribution coefficient by a 2-degree polynomial equation. The permeation profile and related kinetic parameters of terbutaline sulfate was determined in presence of 3 ester-type permeation enhancers incorporated in the films, viz methyl laureate, isopropyl lanolate, and isopropyl myristate. Among the 3, the more pronounced enhancing effect was obtained with isopropyl myristate, regarding the permeation flux, permeability coefficient, and diffusion coefficient. This was attributed to solubility parameter of isopropyl myristate being closer to the solubility parameter of human skin, and such a pronounced enhancing effect was probably caused by its passage across the skin barrier through the lipid pathway. PMID- 16353975 TI - Pellet manufacturing by extrusion-spheronization using process analytical technology. AB - The aim of this study was to investigate the phase transitions occurring in nitrofurantoin and theophylline formulations during pelletization by extrusion spheronization. An at-line process analytical technology (PAT) approach was used to increase the understanding of the solid-state behavior of the active pharmaceutical ingredients (APIs) during pelletization. Raman spectroscopy, near infrared (NIR) spectroscopy, and X-ray powder diffraction (XRPD) were used in the characterization of polymorphic changes during the process. Samples were collected at the end of each processing stage (blending, granulation, extrusion, spheronization, and drying). Batches were dried at 3 temperature levels (60 degrees C, 100 degrees C, and 135 degrees C). Water induced a hydrate formation in both model formulations during processing. NIR spectroscopy gave valuable real time data about the state of water in the system, but it was not able to detect the hydrate formation in the theophylline and nitrofurantoin formulations during the granulation, extrusion, and spheronization stages because of the saturation of the water signal. Raman and XRPD measurement results confirmed the expected pseudopolymorphic changes of the APIs in the wet process stages. The relatively low level of Raman signal with the theophylline formulation complicated the interpretation. The drying temperature had a significant effect on dehydration. For a channel hydrate (theophylline), dehydration occurred at lower drying temperatures. In the case of isolated site hydrate (nitrofurantoin), dehydration was observed at higher temperatures. To reach an understanding of the process and to find the critical process parameters, the use of complementary analytical techniques are absolutely necessary when signals from APIs and different excipients overlap each other. PMID- 16353976 TI - Insulin gel as an alternate to parenteral insulin: formulation, preclinical, and clinical studies. AB - The objective of the present study was to formulate insulin gel for intranasal administration and to evaluate with respect to in vitro release studies and hypoglycemic activity in animal model and healthy human volunteers. The insulin gel was formulated using the combination of carbopol and hydroxypropyl methylcellulose as gelling agent. The in vivo efficacy of insulin gel administered intranasally was assessed by measuring the blood glucose levels and serum insulin levels at specified time intervals in rats and humans. The use of bioadhesive nasal gel containing insulin not only promoted the prolonged contact between the drug and the absorptive sites in the nasal cavity but also facilitated direct absorption of medicament through the nasal mucosa. Absorption of the drug through the nasal mucosa was high in the first 0.5 to 1.5 hours of the study with a sharp decline in blood sugar and rise in insulin values corresponding to that decline in blood sugar. This study further demonstrates that administration of insulin intranasally in gel form is a pleasant and painless alternative to injectable insulin. PMID- 16353977 TI - Effect of polysulfonate resins and direct compression fillers on multiple-unit sustained-release dextromethorphan resinate tablets. AB - The purpose of this work was to investigate the effect of different polysulfonate resins and direct compression fillers on physical properties of multiple-unit sustained-release dextromethorphan (DMP) tablets. DMP resinates were formed by a complexation of DMP and strong cation exchange resins, Dowex 50 W and Amberlite IRP69. The tablets consisted of the DMP resinates and direct compression fillers, such as microcrystalline cellulose (MCC), dicalcium phosphate dihydrate (DCP), and spray-dried rice starch (SDRS). Physical properties of tablets, such as hardness, disintegration time, and in vitro release, were investigated. A good performance of the tablets was obtained when MCC or SDRS was used. The use of rod like and plate-like particles of Amberlite IRP69 caused a statistical decrease in tablet hardness, whereas good tablet hardness was obtained when spherical particle of Dowex 50 W was used. The plastic deformation of the fillers, such as MCC and SDRS, caused a little change in the release of DMP. A higher release rate constant was found in the tablets containing DCP and Dowex 50 W, indicating the fracture of the resinates under compression, which was attributable to the fragmentation of DCP. However, the release of DMP from the tablets using Amberlite IRP69 was not significantly changed because of the higher degree of cross-linking of the resinates, which exhibited more resistance to deformation under compression. In conclusion, the properties of polysulfonate resin, such as particle shape and degree of cross-linking, and the deformation under compaction of fillers affect the physical properties and the drug release of the resinate tablets. PMID- 16353978 TI - Monitoring the fluidized bed granulation process based on S-statistic analysis of a pressure time series. AB - Pressure fluctuation measurements collected during the fluidized bed granulation of pharmaceutical granule have been analyzed using the attractor comparison technique denoted as the S-statistic. Divergence of the bed state from the reference during granulation is followed by a return to a condition statistically similar to the original state of the dry fluidized ingredients on drying. This suggests insensitivity of the S-statistic technique to the changes in particle size distribution occurring during the granulation process. Consequently, the monitoring of pressure fluctuations alone may provide an easily implemented technique for the tracking of granule moisture and process end-point determination. PMID- 16353979 TI - Preparation and characteristics of high-amylose corn starch/pectin blend microparticles: a technical note. PMID- 16353980 TI - Papain entrapment in alginate beads for stability improvement and site-specific delivery: physicochemical characterization and factorial optimization using neural network modeling. AB - This work examines the influence of various process parameters (like sodium alginate concentration, calcium chloride concentration, and hardening time) on papain entrapped in ionotropically cross-linked alginate beads for stability improvement and site-specific delivery to the small intestine using neural network modeling. A 3(3) full-factorial design and feed-forward neural network with multilayer perceptron was used to investigate the effect of process variables on percentage of entrapment, time required for 50% and 90% of the enzyme release, particle size, and angle of repose. Topographical characterization was conducted by scanning electron microscopy, and entrapment was confirmed by Fourier transform infrared spectroscopy and differential scanning calorimetry. Times required for 50% (T(50)) and 90% (T(90)) of enzyme release were increased in all 3 of the process variables. Percentage entrapment and particle size were found to be directly proportional to sodium alginate concentration and inversely proportional to calcium chloride concentration and hardening time, whereas angle of repose and degree of cross-linking showed exactly opposite proportionality. Beads with >90% entrapment and T(50) of <10 minutes could be obtained at the low levels of all 3 of the process variables. The inability of beads to dissolve in acidic environment, with complete dissolution in buffer of pH >or=6.8, showed the suitability of beads to release papain into the small intestine. The shelf-life of the capsules prepared using the papain-loaded alginate beads was found to be 3.60 years compared with 1.01 years of the marketed formulation. It can be inferred from the above results that the proposed methodology can be used to prepare papain-loaded alginate beads for stability improvement and site-specific delivery. PMID- 16353981 TI - Devitrification of amorphous celecoxib. AB - The purpose of this research was to analyze the devitrification of amorphous celecoxib (CEL) in the presence of different stressors (temperature, pressure, and/or humidity) encountered during processing of solid dosage forms. Amorphous CEL was prepared in situ in the analytical instruments, as well as in laboratory, by quench-cooling of melt process, and analyzed by dynamic mechanical thermal analysis, differential scanning calorimetry, microscopy, and Fourier-transform infrared spectroscopy. Amorphous CEL prepared in situ in the analytical instruments was resistant to crystallization under the influence of temperature and/or pressure, because of its protection from the external environment during preparation. These samples exhibited structural relaxation during annealing at 25 degrees C/0% relative humidity (RH) for 16 hours. Generation of amorphous CEL in the laboratory resulted in partially crystalline samples, because of exposure to environmental temperature and humidity, resulting in incomplete vitrification. Subjection to thermal stress favored crystallization of amorphous CEL into metastable polymorphic forms, which were not obtained by solvent recrystallization approach. Temperature and humidity were identified as the major factors promoting devitrification of amorphous CEL, leading to loss of solubility advantage. Exposure to International Conference on Harmonization-specified accelerated stability storage conditions (40 degrees C/75% RH) resulted in complete devitrification of amorphous CEL within 15 days. The phase transformation process of amorphous CEL along the temperature scale was examined visually, as well as spectrally. This propensity for devitrification of amorphous CEL seemed to depend on the strength of differential molecular interactions between the amorphous and crystalline form. PMID- 16353982 TI - Controlled release of drug via methylcellulose-carboxyvinylpolymer interpolymer complex solid dispersion. AB - The purpose of this research was to examine the controlled release of phenacetin (PHE) from solid dispersion by the formation of an interpolymer complex between methylcellulose (MC) and carboxyvinylpolymer (CP). The PHE/polymer composition ratio was fixed at 20:80 (w/w) in the solid dispersion. The effect of the MC/CP ratio and molecular weight of MC on the PHE release was studied. The release of PHE from the solid-dispersion granules depended on the MC/CP ratio, with a ratio of 50:50 giving the lowest rate of release. In aqueous solution, this MC/CP ratio resulted in the lowest transmittance, suggesting a maximal extent of interpolymer complex formation between MC and CP. Furthermore, at a MC/CP ratio of 50:50, the release of PHE from the solid dispersion granules decreased as the molecular weight of the MC increased, reaching a plateau at molecular weights >or=180,000. The contributions of diffusion and polymer relaxation to PHE release increased as the molecular weight of the MC increased. This study shows that it is feasible to control the release of PHE from MC-CP solid dispersion granules by modulating complex formation between MC and CP, which can be accomplished by altering the MC/CP ratio and the molecular weight of MC. PMID- 16353983 TI - Movement of different-shaped particles in a pan-coating device using novel video imaging techniques. AB - The purpose of this study was to investigate the effects of particle shape on the movement of particles in a pan-coating device using novel video-imaging techniques. An area scan CCD camera was installed inside a 24-in pan coater at the same location as that of a spray nozzle, and the movement of particles was tracked using machine vision. A white tracer particle was introduced inside a bed of black-coated particles. The effects of pan loading, pan speed, and particle shape on the movement of particles was studied. The response variables were circulation time, surface time, projected area of particle per pass, dynamic angle of repose, cascading velocity, and dispersion coefficient. Experiments were conducted at 3 different pan speeds, 6, 9, and 12 rpm, and 2 fill levels (ratio of bed depth to pan diameter), one eighth and one quarter, and data were collected over a 30-minute time period. The differences in circulation times of spheres and tablets, with similar volume equivalent diameter as that of the sphere, were found to be insignificant at the 95% confidence interval. The circulation time ranged from 2.8 to 10.8 seconds depending on the operating condition and increased with increasing pan load and decreasing pan speed. The distributions of circulation time, surface time, and projected surface area were found to be nonnormal. The dynamic angle of repose for tablets was higher than for spheres. Also, the bed surface for spheres was much flatter in comparison with tablets where the bed shape attained a "wave-like" form. The average velocity of tablets in the cascading layer was found to be significantly higher than spheres. A linear model (R(2) > 0.98) best described the variation of velocity as a function of pan speed for all of the operating conditions. PMID- 16353984 TI - Quantifying crystal form content in physical mixtures of (+/-)-tartaric acid and (+)-tartaric acid using near infrared reflectance spectroscopy. AB - The objective of this study was to use diffuse reflectance near infrared spectroscopy (NIRS) to determine racemic compound content in physical mixtures composed primarily of the enantiomorph and to assess the error, instrument reproducibility and limits of detection (LOD) and quantification (LOQ) of the method. Physical mixtures ranging from 0 to 25% (+/-)-tartaric acid in (+) tartaric acid were prepared and spectra of the powder samples contained in glass vials were obtained using a Foss NIRSystems Model 5000 monochrometer equipped with a Rapid Content Analyzer scanning from 1100 to 2500 nm. A calibration curve was constructed by plotting (+/-)-tartaric acid weight percent against the 2(nd) derivative values of log (1/R) vs lambda at a single wavelength, normalized with a denominator wavelength (1480 nm/1280 nm). Excellent linearity was observed (R(2)=0.9999). The standard error of calibration (SEC) was 0.07 and the standard error of prediction (SEP) for the validation set was 0.11. Instrument and method errors for samples in the 2% composition range ((+/-)-tartaric acid in (+) tartaric acid) were less than 1% RSD and 3% RSD, respectively. The practical LOD and LOQ were 0.1% and 0.5%, respectively, and comparable to the calculated LOD and LOQ. These studies show that NIRS can be used as a rapid and sensitive quantitative method for determining racemate content in the presence of the enatiomerically pure crystal in the solid-state. PMID- 16353985 TI - The effects of physical parameters on laser-induced breakdown spectroscopy analysis of intact tablets. AB - With the advent of the Food and Drug Administration initiatives to investigate and encourage the use of process analytical technologies, laser-induced breakdown spectroscopy (LIBS) is considered an excellent analytical tool to understand the processability of solid dosage form. In this article, the feasibility of the LIBS system for quantitation of active drug within a solid dosage form, as well as the effects of various physical parameters on its signal, is investigated. A model drug containing chlorine and sulfur was used. The examination of the specificity and reproducibility of the measurements led to the use of chlorine and carbon as the internal standard. An overall relative SD of 1.1% for the signal was found. For quantitation purposes, calibration curves using compound-X in formulated tablets were generated. It was found that curves generated from roller-compaction tablets generally gave higher LIBS signal than those generated using direct compressed (DC) process. To investigate these differences, effect of LIBS signals from several physical properties of the tablets were examined. It was found that unmilled compound-X used in the manufacture of the tablets gave a LIBS signal 30% lower than when milled compound-X was used. However, by using multiple crushing recompression DC process of the milled compound-X, the LIBS results were comparable with those found from both processed tablets using milled compound-X. Other physical parameters, such as wide ranges of granule size and tablet hardness found in the typical manufacturing process, had limited effect on the LIBS signal. From these results, it was noted that for accurate quantitation, it is necessary to use the same physical properties of compound-X and the same manufacturing process in the calibration standards as the actual samples. PMID- 16353986 TI - Process analytical technology case study part I: feasibility studies for quantitative near-infrared method development. AB - This article is the first of a series of articles detailing the development of near-infrared (NIR) methods for solid-dosage form analysis. Experiments were conducted at the Duquesne University Center for Pharmaceutical Technology to qualify the capabilities of instrumentation and sample handling systems, evaluate the potential effect of one source of a process signature on calibration development, and compare the utility of reflection and transmission data collection methods. A database of 572 production-scale sample spectra was used to evaluate the interbatch spectral variability of samples produced under routine manufacturing conditions. A second database of 540 spectra from samples produced under various compression conditions was analyzed to determine the feasibility of pooling spectral data acquired from samples produced at diverse scales. Instrument qualification tests were performed, and appropriate limits for instrument performance were established. To evaluate the repeatability of the sample positioning system, multiple measurements of a single tablet were collected. With the application of appropriate spectral preprocessing techniques, sample repositioning error was found to be insignificant with respect to NIR analyses of product quality attributes. Sample shielding was demonstrated to be unnecessary for transmission analyses. A process signature was identified in the reflection data. Additional tests demonstrated that the process signature was largely orthogonal to spectral variation because of hardness. Principal component analysis of the compression sample set data demonstrated the potential for quantitative model development. For the data sets studied, reflection analysis was demonstrated to be more robust than transmission analysis. PMID- 16353987 TI - Process analytical technology case study: part II. Development and validation of quantitative near-infrared calibrations in support of a process analytical technology application for real-time release. AB - This article is the second of a series of articles detailing the development of near-infrared (NIR) methods for solid dosage-form analysis. Experiments were conducted at the Duquesne University Center for Pharmaceutical Technology to demonstrate a method for developing and validating NIR models for the analysis of active pharmaceutical ingredient (API) content and hardness of a solid dosage form. Robustness and cross-validation testing were used to optimize the API content and hardness models. For the API content calibration, the optimal model was determined as multiplicative scatter correction with Savitsky-Golay first derivative preprocessing followed by partial least-squares (PLS) regression including 4 latent variables. API content calibration achieved root mean squared error (RMSE) and root mean square error of cross validation (RMSECV) of 1.48 and 1.80 mg, respectively. PLS regression and baseline-fit calibration models were compared for the prediction of tablet hardness. Based on robustness testing, PLS regression was selected for the final hardness model, with RMSE and RMSECV of 8.1 and 8.8 N, respectively. Validation testing indicated that API content and hardness of production-scale tablets is predicted with root mean square error of prediction of 1.04 mg and 8.5 N, respectively. Explicit robustness testing for high-flux noise and wavelength uncertainty demonstrated the robustness of the API concentration calibration model with respect to normal instrument operating conditions. PMID- 16353988 TI - Process analytical technology case study, part III: calibration monitoring and transfer. AB - This is the third of a series of articles detailing the development of near infrared spectroscopy methods for solid dosage form analysis. Experiments were conducted at the Duquesne University Center for Pharmaceutical Technology to develop a system for continuous calibration monitoring and formulate an appropriate strategy for calibration transfer. Indicators of high-flux noise (noise factor level) and wavelength uncertainty were developed. These measurements, in combination with Hotelling's T(2) and Q residual, are used to continuously monitor instrument performance and model relevance. Four calibration transfer techniques were compared. Three established techniques, finite impulse response filtering, generalized least squares weighting, and piecewise direct standardization were evaluated. A fourth technique, baseline subtraction, was the most effective for calibration transfer. Using as few as 15 transfer samples, predictive capability of the analytical method was maintained across multiple instruments and major instrument maintenance. PMID- 16353989 TI - Lecithin organogels as a potential phospholipid-structured system for topical drug delivery: a review. AB - The purpose of this review is to give an insight into the considerable potential of lecithin organogels (LOs) in the applications meant for topical drug delivery. LOs are clear, thermodynamically stable, viscoelastic, and biocompatible jelly like phases, chiefly composed of hydrated phospholipids and appropriate organic liquid. These systems are currently of interest to the pharmaceutical scientist because of their structural and functional benefits. Several therapeutic agents have been formulated as LOs for their facilitated transport through topical route (for dermal or transdermal effect), with some very encouraging results. The improved topical drug delivery has mainly been attributed to the biphasic drug solubility, the desired drug partitioning, and the modification of skin barrier function by the organogel components. Being thermodynamically stable, LOs are prepared by spontaneous emulsification and therefore possess prolonged shelf life. The utility of this novel matrix as a topical vehicle has further increased owing to its very low skin irritancy potential. Varied aspects of LOs viz formation, composition, phase behavior, and characterization have been elaborated, including a general discussion on the developmental background. Besides a comprehensive update on the topical applications of lecithin organogels, the review also includes a detailed account on the mechanistics of organogelling. PMID- 16353990 TI - Excipient selection can significantly affect solid-state phase transformation in formulation during wet granulation. AB - Phase transformations in formulations can lead to instability in physicochemical, biopharmaceutical, and processing properties of products. The influences of formulation design on the optimal dosage forms should be specified. The aim here was to investigate whether excipients with different water sorption behavior affect hydrate formation of nitrofurantoin in wet masses. Nitrofurantoin anhydrate was used as a hydrate-forming model drug, and 4 excipients with different water-absorbing potential (amorphous low-substituted hydroxypropylcellulose, modified maize starch, partially amorphous silicified microcrystalline cellulose, and crystalline alpha-lactose monohydrate) were granulated with varying amounts of purified water. Off-line evaluation of wet masses containing nitrofurantoin anhydrate and excipient (1:1) was performed using an X-ray powder diffractometer (XRPD) and near-infrared spectroscopy, and drying phase was evaluated by variable temperature XRPD. Only amorphous excipient in the formulation retarded hydrate formation of an active pharmaceutical ingredient (API) at high water contents. Hygroscopic partially crystalline excipient hindered hydrate formation of API at low water contents. Crystalline excipient was unable to control hydrate formation of API. The character of excipient affects the stability of formulation. Thus, correct selection of excipients for the formulation can control processing-induced phase transitions and improve the storage stability of the final dosage form. PMID- 16353991 TI - Development of a dialysis in vitro release method for biodegradable microspheres. AB - The purpose of this research was to develop a simple and convenient in vitro release method for biodegradable microspheres using a commercially available dialyzer. A 25 KD MWCO Float-a-Lyzer was used to evaluate peptide diffusion at 37 degrees C and 55 degrees C in different buffers and assess the effect of peptide concentration. In vitro release of Leuprolide from PLGA microspheres, having a 1 month duration of action, was assessed using the dialyzer and compared with the commonly used sample and separate method with and without agitation. Peptide diffusion through the dialysis membrane was rapid at 37 degrees C and 55 degrees C in all buffers and was independent of peptide concentration. There was no detectable binding to the membrane under the conditions of the study. In vitro release of Leuprolide from PLGA microspheres was tri-phasic and was complete in 28 days with the dialysis technique. With the sample and separate technique, linear release profiles were obtained with complete release occurring under conditions of agitation. Diffusion through the dialysis membrane was sufficiently rapid to qualify the Float-a-Lyzer for an in vitro release system for microparticulate dosage forms. Membrane characteristics render it useful to study drug release under real-time and accelerated conditions. PMID- 16353993 TI - Dry granulation and compression of spray-dried plant extracts. AB - The purpose of this research was to evaluate the influence of dry granulation parameters on granule and tablet properties of spray-dried extract (SDE) from Maytenus ilicifolia, which is widely used in Brazil in the treatment of gastric disorders. The compressional behavior of the SDE and granules of the SDE was characterized by Heckel plots. The tablet properties of powders, granules, and formulations containing a high extract dose were compared. The SDE was blended with 2% magnesium stearate and 1% colloidal silicon dioxide and compacted to produce granules after slugging or roll compaction. The influences of the granulation process and the roll compaction force on the technological properties of the granules were studied. The flowability and density of spray-dried particles were improved after granulation. Tablets produced by direct compression of granules showed lower crushing strength than the ones obtained from nongranulated material. The compressional analysis by Heckel plots revealed that the SDE undergoes plastic deformation with a very low tendency to rearrangement at an early stage of compression. On the other hand, the granules showed an intensive rearrangement as a consequence of fragmentation and rebounding. However, when the compaction pressure was increased, the granules showed plastic deformation. The mean yield pressure values showed that both granulation techniques and the roll compaction force were able to reduce the material's ability to undergo plastic deformation. Finally, the tablet containing a high dose of granules showed a close dependence between crushing strength and the densification degree of the granules (ie, roll compaction force). PMID- 16353994 TI - Optimization of tablets containing a high dose of spray-dried plant extract: a technical note. PMID- 16353992 TI - Cyclodextrins in drug delivery: an updated review. AB - The purpose of this review is to discuss and summarize some of the interesting findings and applications of cyclodextrins (CDs) and their derivatives in different areas of drug delivery, particularly in protein and peptide drug delivery and gene delivery. The article highlights important CD applications in the design of various novel delivery systems like liposomes, microspheres, microcapsules, and nanoparticles. In addition to their well-known effects on drug solubility and dissolution, bioavailability, safety, and stability, their use as excipients in drug formulation are also discussed in this article. The article also focuses on various factors influencing inclusion complex formation because an understanding of the same is necessary for proper handling of these versatile materials. Some important considerations in selecting CDs in drug formulation such as their commercial availability, regulatory status, and patent status are also summarized. CDs, because of their continuing ability to find several novel applications in drug delivery, are expected to solve many problems associated with the delivery of different novel drugs through different delivery routes. PMID- 16353995 TI - Floating drug delivery systems: a review. AB - The purpose of writing this review on floating drug delivery systems (FDDS) was to compile the recent literature with special focus on the principal mechanism of floatation to achieve gastric retention. The recent developments of FDDS including the physiological and formulation variables affecting gastric retention, approaches to design single-unit and multiple-unit floating systems, and their classification and formulation aspects are covered in detail. This review also summarizes the in vitro techniques, in vivo studies to evaluate the performance and application of floating systems, and applications of these systems. These systems are useful to several problems encountered during the development of a pharmaceutical dosage form. PMID- 16353996 TI - Production of alkaline protease with immobilized cells of Bacillus subtilis PE-11 in various matrices by entrapment technique. AB - The purpose of this investigation was to study the effect of Bacillus subtilis PE 11 cells immobilized in various matrices, such as calcium alginate, k Carrageenan, ployacrylamide, agar-agar, and gelatin, for the production of alkaline protease. Calcium alginate was found to be an effective and suitable matrix for higher alkaline protease productivity compared to the other matrices studied. All the matrices were selected for repeated batch fermentation. The average specific volumetric productivity with calcium alginate was 15.11 U/mL/hour, which was 79.03% higher production over the conventional free-cell fermentation. Similarly, the specific volumetric productivity by repeated batch fermentation was 13.68 U/mL/hour with k-Carrageenan, 12.44 U/mL/hour with agar agar, 11.71 U/mL/hour with polyacrylamide, and 10.32 U/mL/hour with gelatin. In the repeated batch fermentations of the shake flasks, an optimum level of enzyme was maintained for 9 days using calcium alginate immobilized cells. From the results, it is concluded that the immobilized cells of B subtilis PE-11 in calcium alginate are more efficient for the production of alkaline protease with repeated batch fermentation. The alginate immobilized cells of B subtilis PE-11 can be proposed as an effective biocatalyst for repeated usage for maximum production of alkaline protease. PMID- 16353997 TI - Application of powder rheometer to determine powder flow properties and lubrication efficiency of pharmaceutical particulate systems. AB - The objective of this study was to understand the behavior of particulate systems under different conditions of shear dynamics before and after granulation and to investigate the efficiency of powder lubrication. Three drug powders, metronidazole, colloidal bismuth citrate, and tetracycline hydrochloride, were chosen as model drugs representing noncohesive and cohesive powder systems. Each powder was individually granulated with microcrystalline cellulose and 5%PVP as a binder. One portion from each granulation was lubricated with different levels of magnesium stearate for 5 minutes. The powder characterization was performed on the plain powders, nonlubricated and lubricated granules using powder rheometer equipped with a helical blade rotating and moving under experimentally fixed set of parameters. The profiles of interaction during the force-distance measurements indicate that powder compresses, expands, and shears many times in a test cycle. Test profiles also clearly reveal existence of significant differences between cohesive and noncohesive powders. In all cases lubrication normalized the overall interactive nature of the powder by reducing peaks and valleys as observed from the profiles and reduced the frictional effect. The developed methods are easy to perform and will allow formulation scientists to better understand powder behavior and help in predicting potential impact of processing factors on particulate systems. PMID- 16353998 TI - Preparation and characterization of etoricoxib solid dispersions using lipid carriers by spray drying technique. AB - The basic objectives of this study were to prepare and characterize solid dispersions of poorly water-soluble drug etoricoxib using lipid carriers by spray drying technique. The properties of solid dispersions were studied by diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), differential scanning calorimetry (DSC), hot-stage microscopy (HSM), radiograph powder diffraction (XRPD), and dissolution studies. The absence of etoricoxib peaks in XRPD profiles of solid dispersions suggests the transformation of crystalline etoricoxib into an amorphous form. In the HSM examination of solid dispersions, the dissolution of drug in the lipid carriers was observed, which was also confirmed by the absence of etoricoxib peak in DSC curves of solid dispersions. The DRIFTS spectra revealed the presence of hydrogen bonding in solid dispersions. The in vitro dissolution test showed a significant increase in the dissolution rate of solid dispersions as compared with pure etoricoxib, spray dried etoricoxib, and physical mixtures of drug with lipid carriers. Therefore, the dissolution rate of poorly water-soluble drug etoricoxib can be significantly enhanced by the preparation of solid dispersions using lipid carriers by spray drying technique. PMID- 16353999 TI - Physical changes of beta-sitosterol crystals in oily suspensions during heating. AB - The aim of this research was to describe the thermal behavior of beta-sitosterol crystals in oil-suspensions with a focus on the role of water during heating. The suspensions were prepared by recrystallization in order to achieve a microcrystalline particle size. The structural changes together with the mechanical properties of the suspensions during heating were studied by using variable temperature X-ray powder diffractometry (VT-XRPD), differential scanning calorimetry (DSC), and dynamic mechanical analysis (DMA). Hydrated beta sitosterol crystals in an oil-suspension dehydrated, despite the composition of the suspensions, at low temperatures. At high beta-sitosterol concentration, the monohydrate crystal form changed partially to a hemihydrated form, and when only a small amount of water was initially incorporated, the hemihydrate crystal form dehydrated to a mostly anhydrate crystal form. The released water, which was immiscible in the surrounding oil, caused the recrystallization of hydrated beta sitosterol during cooling. This procedure indicated a reversible dehydration process. Structural and thermal analysis of beta-sitosterol crystals in suspensions, together with mechanical analysis made it possible to understand various physical changes during heating. PMID- 16354000 TI - Supercritical assisted atomization: a novel technology for microparticles preparation of an asthma-controlling drug. AB - The objective of this study was to produce microparticles of a new asthma controlling drug by supercritical assisted atomization (SAA), proposed as an alternative to conventional jet-milling process. SAA is based on the solubilization of supercritical carbon dioxide in a liquid solution containing the drug; the ternary mixture is then sprayed through a nozzle, and microparticles are formed as a consequence of the enhanced atomization. SAA process parameters studied were precipitator temperature, nozzle diameter, and drug concentration in the liquid solution. Their influence was evaluated on morphology and size of precipitated particles. Spherical particles with mean particle size ranging from 1 to 3 microm of the new anti-asthma drug were produced by SAA. The mass median aerodynamic diameter (MMAD) of the SAA micronized particles and of the conventional jet-milled drug was used to compare the results obtainable using the 2 techniques. Particularly, MMADs from 1.6 to 4.0 microm were obtained by SAA at the optimum operating conditions and by varying the concentration of the solution injected. MMAD of 6.0 microm was calculated for the jet-milled drug. SAA samples also exhibited narrower particle size distribution (PSD). A good control of particle size and distribution together with no drug degradation was obtained by SAA process. PMID- 16354001 TI - Controlled release from triple layer, donut-shaped tablets with enteric polymers. AB - The purpose of this research was to evaluate triple layer, donut-shaped tablets (TLDSTs) for extended release dosage forms. TLDSTs were prepared by layering 3 powders sequentially after pressing them with a punch. The core tablet consisted of enteric polymers, mainly hydroxypropyl methylcellulose acetate succinate, and the bottom and top layers were made of a water-insoluble polymer, ethyl cellulose. Drug release kinetics were dependent on the pH of the dissolution medium and the drug properties, such as solubility, salt forms of weak acid and weak base drugs, and drug loading. At a 10% drug loading level, all drugs, regardless of their type or solubility, yielded the same release profiles within an acceptable level of experimental error. As drug loading increased from 10% to 30%, the drug release rate of neutral drugs increased for all except sulfathiazole, which retained the same kinetics as at 10% loading. HCl salts of weak base drugs had much slower release rates than did those of neutral drugs (eg, theophylline) as drug loading increased. The release of labetalol HCl retarded as drug loading increased from 10% to 30%. On the other hand, Na salts of weak acid drugs had much higher release rates than did those of neutral drugs (eg, theophylline). Drug release kinetics were governed by the ionization/erosion process with slight drug diffusion, observing no perfect straight line. A mathematical expression for drug release kinetics (erosion-controlled system) of TLDSTs is presented. In summary, a TLDST is a good design to obtain zero-order or nearly zero-order release kinetics for a wide range of drug solubilities. PMID- 16354002 TI - Studies on spray-dried mixtures of chitosan and hydrolyzed gelatin as tablet binder: a technical note. PMID- 16354003 TI - Solid-state fluorescence of the trihydrate phases of ampicillin and amoxicillin. AB - The purpose of this work was to study the effects of crystal structure on the solid-state photoluminescence of the trihydrate phases of ampicillin and amoxicillin, and to contrast these spectra with analogous spectra obtained on the molecules dissolved in a solution phase. The polymorphic identity of the analytes was established using x-ray powder diffraction and Fourier transform infrared absorption spectroscopy, and the solid-state luminescence spectra obtained under ambient conditions. It was found that the solid-state excitation and emission spectra of ampicillin trihydrate and amoxicillin trihydrate were dominated by energy transfer and exciton effects, which were manifested as decreases in the energy of the excitation and emission bands of the solid-state systems relative to those of the free molecule in solution. The photoluminescence data revealed that in spite of the known structural similarity of ampicillin trihydrate and amoxicillin trihydrate, the magnitude of the Davydov splitting, and the degree of band energy shifting differed between the 2 systems. This finding indicates that the small differences in crystal structure existing between the 2 compounds leads to measurable differences in the patterns of energy transfer. PMID- 16354004 TI - Generalization of a prototype intelligent hybrid system for hard gelatin capsule formulation development. AB - The aim of this project was to expand a previously developed prototype expert network for use in the analysis of multiple biopharmaceutics classification system (BCS) class II drugs. The model drugs used were carbamazepine, chlorpropamide, diazepam, ibuprofen, ketoprofen, naproxen, and piroxicam. Recommended formulations were manufactured and tested for dissolution performance. A comprehensive training data set for the model drugs was developed and used to retrain the artificial neural network. The training and the system were validated based on the comparison of predicted and observed performance of the recommended formulations. The initial test of the system resulted in high error values, indicating poor prediction capabilities for drugs other than piroxicam. A new data set, containing 182 batches, was used for retraining. Ten percent of the test batches were used for cross-validation, resulting in models with R2 > or = 70%. The comparison of observed performance to the predicted performance found that the system predicted successfully. The hybrid network was generally able to predict the amount of drug dissolved within 5% for the model drugs. Through validation, the system was proven to be capable of designing formulations that met specific drug performance criteria. By including parameters to address wettability and the intrinsic dissolution characteristics of the drugs, the hybrid system was shown to be suitable for analysis of multiple BCS class II drugs. PMID- 16354005 TI - Effects of plantain and corn starches on the mechanical and disintegration properties of paracetamol tablets. AB - The effects of plantain starch obtained from the unripe fruit of the plant Musa paradisiaca L. (Musaceae) on the mechanical and disintegration properties of paracetamol tablets have been investigated in comparison with the effects of corn starch BP using a 2(3) factorial experimental design. The individual and combined effects of nature of starch binder (N), concentration of starch binder (C), and the relative density of tablet (RD) on the tensile strength (TS), brittle fracture index (BFI), and disintegration time (DT) of the tablets were investigated. The ranking of the individual effects on TS was RD > C >> N, on BFI was C >> RD > N and on DT was N > C > RD. The ranking for the interaction effects on TS and DT was N-C >> N-RD > C-RD, while that on BFI was N-C >> C-RD > N-RD. Changing nature of starch from a "low" (plantain starch) to a "high" (corn starch) level, increasing the concentration of starch binding agent from 2.5% to 10.0% wt/wt, and increasing relative density of the tablet from 0.80 to 0.90, led to increase in the values of TS and DT, but a decrease in BFI. Thus, tablets containing plantain starch had lower tensile strength and disintegration time values than those containing corn starch, but showed better ability to reduce the lamination and capping tendency in paracetamol tablet formulation. The interaction between N and C was significantly (P < .001) higher than those between N and RD and between C and RD. There is therefore the need to carefully choose the nature (N) and concentration (C) of starch used as binding agent in tablet formulations to obtain tablets of desired bond strength and disintegration properties. Furthermore, plantain starch could be useful as an alternative binding agent to cornstarch, especially where faster disintegration is required and the problems of lamination and capping are of particular concern. PMID- 16354006 TI - Diclofenac-beta-cyclodextrin binary systems: physicochemical characterization and in vitro dissolution and diffusion studies. AB - The aim of this work was to study the influence of beta-cyclodextrin (beta-CD) on the biopharmaceutic properties of diclofenac (DCF). To this purpose the physicochemical characterization of diclofenac-beta-cyclodextrin binary systems was performed both in solution and solid state. Solid phase characterization was performed using differential scanning calorimetry (DSC), powder x-ray diffractometry (XRD), and Fourier transform infrared spectroscopy (FTIR). Phase solubility analyses, and in vitro permeation experiments through a synthetic membrane were performed in solution. Moreover, DCF/beta-CD interactions were studied in DMSO by 1H nuclear magnetic resonance (NMR) spectroscopy. The effects of different preparation methods and drug-to-beta-CD molar ratios were also evaluated. Phase solubility studies revealed 1:1 M complexation of DCF when the freeze-drying method was used for the preparation of the binary system. The true inclusion for the freeze-dried binary system was confirmed by 1H NMR spectroscopy, DSC, powder XRD, and IR studies. The dissolution study revealed that the drug dissolution rate was improved by the presence of CDs and the highest and promptest release was obtained with the freeze-dried binary system. Diffusion experiments through a silicone membrane showed that DCF diffusion was higher from the saturated drug solution (control) than the freeze-dried inclusion complexes, prepared using different DCF-beta-CD molar ratios. However, the presence of the inclusion complex was able to stabilize the system giving rise to a more regular diffusion profile. PMID- 16354008 TI - A preliminary pharmacokinetic study of liposomal leuprolide dry powder inhaler: a technical note. PMID- 16354007 TI - Lipid synthesis inhibitors: effect on epidermal lipid conformational changes and percutaneous permeation of levodopa. AB - A combination of lipid synthesis inhibitors was used to enhance the in vitro and in vivo permeation of levodopa (LD) across rat epidermis, and their influence on epidermal lipids was investigated using attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. Rat epidermis was treated with ethanol and a combination of atorvastatin (750 microg/7 cm2), cerulenin (20 microg/7 cm2), and beta-chloroalanine (600 microg/7 cm2) for sustaining the reduced content of epidermal cholesterol, fatty acids (as triglycerides), and ceramide (as sphingosine), respectively, in viable rat skin. This treatment resulted in significant (P < .05) synthesis inhibition of skin lipids up to 48 hours and 6-fold enhancement in the in vitro permeation of LD. The effective plasma concentration of LD was achieved within 1 hour and maintained over 48 hours after topical application to rat epidermis treated with a combination of these lipid synthesis inhibitors. ATR-FTIR studies of inhibitor(s)-treated rat epidermis revealed a significant decrease (P < .05) in peak height and area for both asymmetric and symmetric C-H stretching absorbances, suggesting extraction of lipids. However, an insignificant (P < .05) shift in the frequency of these peaks suggested no fluidization of epidermal lipids by lipid synthesis inhibitors. A direct correlation was observed between epidermal lipid synthesis inhibition, decrease in peak height or area, and percutaneous permeation of LD. Skin lipid synthesis inhibition by a combination of lipid synthesis inhibitors seems to offer a feasible approach for enhancing the transcutaneous delivery of LD. PMID- 16354009 TI - Preparation, in vitro release, in vivo absorption and biocompatibility studies of insulin-loaded microspheres in rabbits. AB - The purpose of this study was to develop a single-dose insulin delivery system based on poly (lactide-co-glycolide) (PLGA) microspheres to provide basal insulin level for a prolonged period. Insulin-loaded PLGA microspheres were prepared by water-in-oil-in-water double emulsion (batch A) and solid-in-oil-in-water emulsion (batch B) methods. Microspheres were characterized for physical characteristics and in vitro release. In vivo absorption of insulin and biocompatibility of insulin-loaded PLGA microspheres were performed in diabetic New Zealand white rabbits. Light and transmission electron microscopy were performed on the skin tissues excised from microspheres injected sites in order to study the biocompatibility. The burst release of insulin was high (47%) from batch B and low (5%) from batch A. Therefore, we mixed microspheres of batch A and B in ratio of 3:1 w/w, which produced desirable in vitro release profile. In vivo absorption study showed that insulin-loaded microspheres provided a serum insulin level of 20-40 microU/ml up to 40 days. Biocompatibility study provided evidence of normal inflammatory and foreign body reactions, which were characterized by the presence of macrophages, fibroblasts and foreign body giant cells. Neither necrosis nor tissue damage was identified. At the end of 12 weeks, no distinct histological differences were observed in comparison to the control tissue samples. In conclusion, insulin-loaded PLGA microspheres controlled the in vivo absorption of insulin to maintain the basal insulin level for longer period and the delivery system was biocompatible. PMID- 16354010 TI - Direct pelletization in a rotary processor controlled by torque measurements. III. Investigation of microcrystalline cellulose and lactose grade. AB - The aim of the present study was to investigate the use of different grades of microcrystalline cellulose (MCC) and lactose in a direct pelletization process in a rotary processor. For this purpose, a mixed 2- and 3-level factorial study was performed to determine the influence of the particle size of microcrystalline cellulose (MCC) (approximately 60 and 105 microm) and lactose (approximately 30, 40, and 55 microm), as well as MCC type (Avicel and Emcocel) on the pelletization process and the physical properties of the prepared pellets. A 1:4 mixture of MCC and lactose was applied, and granulation liquid was added until a 0.45 Nm increase in the torque of the friction plate was reached. All combinations of the 3 factors resulted in spherical pellets of a high physical strength. The particle size of MCC was found to have no marked effect on the amount of water required for agglomerate growth or on the size of the resulting pellets. An increasing particle size of lactose gave rise to more spherical pellets of a more narrow size distribution as well as higher yields. The MCC type was found to affect both the release of the model drug from the prepared pellets and the size distribution. Generally, the determined influence of the investigated factors was small, and direct pelletization in a rotary processor was found to be a robust process, insensitive to variations in the particle size and type of MCC and the particle size of lactose. PMID- 16354011 TI - Effect of polyamidoamine (PAMAM) dendrimers on the in vitro release of water insoluble nifedipine from aqueous gels. AB - The objective of this study was to determine the effect of ethylenediamine core PAMAM dendrimers on the release of nifedipine suspended in aqueous gels and to correlate release to the increase in solubility afforded by the dendrimers. Drug release from aqueous 5% HPMC gels containing nifedipine (2% wt/vol) through 0.2 microm membranes was measured using Enhancer cells and 50% ethanolic solution as the receptor medium. The release from gels containing PAMAM G-3 and G-5 (0.25%-1% wt/vol) was compared with gels containing the cosolvent isopropyl alcohol (10% 80% vol/vol). PAMAM dendrimers significantly increased the solubility of nifedipine. This caused a significant increase in the release rate of nifedipine from the gel suspensions. The increase in drug release depended on the concentration and generation size of the dendrimers added. For higher generations (G-5) lower concentrations were needed to obtain equivalent increases in release. Although the increase in solubility and release was not as high as from gels containing high concentrations of the cosolvent isopropyl alcohol, the dendrimers prevented the recrystallization of the drug that was observed when the gels containing isopropyl alcohol were left open. PMID- 16354012 TI - Proultraflexible lipid vesicles for effective transdermal delivery of levonorgestrel: development, characterization, and performance evaluation. AB - The present investigation aimed at formulation, performance evaluation, and stability studies of new vesicular drug carrier system protransfersomes for transdermal delivery of the contraceptive agent, levonorgestrel. Protransfersome gel (PTG) formulations of levonorgestrel were prepared and characterized for vesicle shape, size, entrapment efficiency, turbidity, and drug permeation across rat skin and were evaluated for their stability. The system was evaluated in vivo for biological assay of progestational activity including endometrial assay, inhibition of the formation of corpora lutea, and weight gain of uterus. The effects of different formulation variables (type of alcohol, type and concentration of surfactant) on transdermal permeability profile were assessed. The optimized PTG formulation showed enhanced in vitro skin permeation flux of 15.82 +/- 0.37 microg/cm2/hr as compared to 0.032 +/- 0.01 microg/cm2/hr for plain drug solution. PTG also showed good stability and after 2 months of storage there was no change in liquid crystalline nature, drug content, and other characteristic parameters. The peak plasma concentration of levonorgestrel (0.139 +/- 0.05 microg/mL) was achieved within 4 hours and maintained until 48 hours by a single topical application of optimized PTG formulation. In vivo performance of the PTG formulation showed increase in the therapeutic efficacy of drug. Results indicated that the optimized protransfersomal formulation of levonorgestrel had better skin permeation potential, sustained release characteristic, and better stability than proliposomal formulation. PMID- 16354013 TI - In vivo ocular availability of ketorolac following ocular instillations of aqueous, oil, and ointment formulations to normal corneas of rabbits: a technical note. PMID- 16354014 TI - Effect of tablet geometrical structure on the dehydration of creatine monohydrate tablets, and their pharmaceutical properties. AB - The effects of compression and pulverization on the dehydration kinetics and hardness of creatine monohydrate tablets were studied using a variety of kinetic equations and physical models. The dehydration behavior of unpulverized and pulverized tablets was investigated by using differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD). The hardness of both unpulverized and pulverized monohydrate tablets was significantly decreased after dehydration. The relationship between the degree of dehydration and the tablet hardness of both unpulverized and pulverized monohydrate tablets formed a straight line. The results suggest that the reduction in tablet hardness is dependent on the dehydration of crystal water, and the values of the slopes indicate that the bonding energy of the unpulverized sample was stronger than that of the pulverized sample. The dehydration kinetics of the unpulverized and pulverized monohydrate tablets were evaluated by analyzing the fit of the isothermal DSC data using a variety of solid-state kinetic models. The dehydration of the unpulverized tablets at various levels of compression pressure followed the 3 dimensional growth of nuclei mechanism. In contrast, although the dehydration kinetics of pulverized monohydrate tablets compressed at 500 and 750 kg/cm2 followed the 3-dimensional diffusion mechanism, those compressed at 1000 kg/cm2 followed the 3-dimensional growth of nuclei mechanism. The PXRD analysis indicated that the diffraction intensity of the pulverized monohydrate powder was significantly lower than that of the unpulverized powder. The diffraction peaks of the (h00) planes and the micropore structure of the unpulverized monohydrate tablets were affected by pulverization and compression force, respectively. PMID- 16354015 TI - Poly(amidoamine) (PAMAM) dendritic nanostructures for controlled site-specific delivery of acidic anti-inflammatory active ingredient. AB - The purpose of the investigation was to evaluate the potential of polyamidoamine (PAMAM) dendrimer as nanoscale drug delivery units for controlled release of water insoluble and acidic anti-inflammatory drug. Flurbiprofen (FB) was selected as a model acidic anti-inflammatory drug. The aqueous solutions of 4.0 generation (G) PAMAM dendrimer in different concentrations were prepared and used further for solubilizing FB. Formation of dendrimer complex was characterized by Fourier transform infrared spectroscopy. The effect of pH on the solubility of FB in dendrimer was evaluated. Dendrimer formulations were further evaluated for in vitro release study and hemolytic toxicity. Pharmacokinetic and biodistribution were studied in male albino rats. Efficacy of dendrimer formulation was tested by carrageenan induced paw edema model. It was observed that the loaded drug displayed initial rapid release (more than 40% till 3rd hour) followed by rather slow release. Pharmacodynamic study revealed 75% inhibition at 4th hour that was maintained above 50% till 8th hour. The mean residence time (MRT) and terminal half-life (THF) of the dendritic formulation increased by 2-fold and 3-fold, respectively, compared with free drug. Hence, with dendritic system the drug is retained for longer duration in the biosystem with 5-fold greater distribution. It may be concluded that the drug-loaded dendrimers not only enhanced the solubility but also controlled the delivery of the bioactive with localized action at the site of inflammation. PMID- 16354018 TI - Autobiography of Jack Simons. PMID- 16354020 TI - List of publications by Jack Simons 1969 through 2005. PMID- 16354016 TI - Kinetic spectrophotometric method for the determination of ramipril in pharmaceutical formulations. AB - The objective of this research was to develop a kinetic spectrophotometric method for determination of ramipril in pure form and pharmaceutical formulations. The method was based on the reaction of carboxylic acid group of the drug with a mixture of potassium iodate (KIO3) and potassium iodide (KI) in aqueous medium at room temperature. The reaction is followed spectrophotometrically by measuring the increase in absorbance at 352 nm as a function of time. The initial-rate and fixed-time methods were adopted for constructing the calibration curves. Both the calibration curves were linear in the concentration range of 10.0-70.0 microg mL( 1). The detection limits were 0.02 microg mL(-1) and 0.15-microg mL(-1) for initial rate and fixed time methods, respectively. The proposed methods are validated statistically and through recovery studies. The point and interval hypothesis tests have been performed confirming that there is no significant difference between the proposed methods and the reference method. The experimental true bias of all samples is less than +/- 2%. The methods have been successfully applied to the determination of ramipril in tablets and capsules. PMID- 16354021 TI - Nucleation from a supercooled binary mixture studied by crossed polarizers. AB - Polarized light passing through a supercooled binary liquid mixture sample is analyzed during the moment of the nucleation of the crystal phase to determine whether the stable equilibrium crystal is nucleated, or whether a transient phase of different composition or broken-symmetry is formed. This experiment is performed for the particular case of heterogeneous nucleation of a supercooled clathrate-forming liquid mixture, tetrahydrofuran (THF)/water, compared with ice nucleating from pure supercooled water. The new experimental results are consistent with the hypothesis that the equilibrium clathrate hydrate crystal is nucleated directly, with no transient phase detected on the time scale of these experiments. PMID- 16354022 TI - Monoligated monovalent Ni: the 3d(Ni)9 manifold of states of NiCu and comparison to the 3d9 States of AlNi, NiH, NiCl, and NiF. AB - A dispersed fluorescence investigation of the low-lying electronic states of NiCu has allowed the observation of four out of the five states that derive from the 3d(Ni)9 3d(Cu)10 sigma2 manifold. Vibrational levels of the ground X2delta(5/2) state corresponding to v = 0-11 are observed and are fit to provide omega(e) = 275.93 +/- 1.06 cm(-1) and omega(e)x(e) = 1.44 +/- 0.11 cm(-1). The v = 0 levels of the higher lying states deriving from the 3d(Ni)9 3d(Cu)10 sigma2 manifold are located at 912, 1466, and 1734 cm(-1), and these states are assigned to omega values of 3/2, 1/2, and 3/2, respectively. The last of these assignments is based on selection rules and is unequivocal; the first two are based on a comparison to ab initio and ligand field calculations and could conceivably be in error. It is also possible that the v = 0 level of the state found at 912 cm(-1) is not observed, so that T0 for the lowest excited state actually lies near 658 cm(-1). These results are modeled using a matrix Hamiltonian based on the existence of a ground manifold of states deriving from the 3d9 configuration on nickel. This matrix Hamiltonian is also applied to the spectroscopically well-known molecules AlNi, NiH, NiCl, and NiF. The term energies of the 2sigma+, 2pi, and 2delta states of these molecules, which all derive from a 3d9 configuration on the nickel atom, display a clear and understandable trend as a function of the electronegativity of the ligands. PMID- 16354023 TI - Sequential bond energies of Fe+ (CO2)n, n = 1-5, determined by threshold collision-induced dissociation and ab initio theory. AB - Collision-induced dissociation of the Fe+ (CO2)n complexes for n = 1-5 is studied using kinetic energy dependent guided ion beam mass spectrometry. In all cases, the primary products are endothermic loss of an intact neutral ligand from the complex. The cross section thresholds are interpreted to yield 0 K bond energies after accounting for the effects of multiple ion-molecule collisions, internal energy of the complexes, and unimolecular decay rates. These values are compared with density functional theoretical values for all five complexes. Theory provides bond energies in reasonable agreement with experiment for n = 1-4 and predictions for the infrared spectroscopy of these complexes that agree nicely with experimental results of Gregoire and Duncan (J. Chem. Phys. 2002, 117, 2120). Our thermochemical results are also compared with the Fe+ (CO)n and Fe+ (N2)n complexes, previously studied. PMID- 16354024 TI - Direct dynamics trajectory study of the reaction of formaldehyde cation with D2: vibrational and zero-point energy effects on quasiclassical trajectories. AB - Quasiclassical, direct dynamics trajectories have been used to study the reaction of formaldehyde cation with molecular hydrogen, simulating the conditions in an experimental study of H2CO+ vibrational effects on this reaction. Effects of five different H2CO+ modes were probed, and we also examined different approaches to treating zero-point energy in quasiclassical trajectories. The calculated absolute cross-sections are in excellent agreement with experiments, and the results provide insight into the reaction mechanism, product scattering behavior, and energy disposal, and how they vary with impact parameter and reactant state. The reaction is sharply orientation-dependent, even at high collision energies, and both trajectories and experiment find that H2CO+ vibration inhibits reaction. On the other hand, the trajectories do not reproduce the anomalously strong effect of nu2(+) (the CO stretch). The origin of the discrepancy and approaches for minimizing such problems in quasiclassical trajectories are discussed. PMID- 16354025 TI - Chemical bonding in Si5(2-) and NaSi5(-) via photoelectron spectroscopy and ab initio calculations. AB - Photoelectron spectroscopy and ab initio calculations are used to investigate the electronic structure and chemical bonding of Si5(-) and Si5(2-) in NaSi5(-). Photoelectron spectra of Si5(-) and NaSi5(-) are obtained at several photon energies and are compared with theoretical calculations at four different levels of theory, TD-B3LYP, R(U)OVGF, UCCSD(T), and EOM-CCSD(T), all with 6-311+G(2df) basis sets. Excellent agreement is observed between experiment and theory, confirming the obtained ground-state structures for Si5(-) and Si5(2-), which are both found to be trigonal bipyramid with D3h symmetry at several levels of theory. Chemical bonding in Si5, Si5(-), and Si5(2-) is analyzed using NPA, molecular orbitals, ELF, and NICS indices. The bonding in Si5(2-) is compared with that in the isoelectronic and isostructural B5H5(2-) species, but they are found to differ due to the involvement of electron densities, which are supposed to be lone pairs in the skeletal bonding in Si5(2-). PMID- 16354026 TI - Temperature-dependent photoelectron spectroscopy of methyl benzoate anions: observation of steric effect in o-methyl benzoate. AB - Temperature-dependent photoelectron spectra of benzoate anion (C6H5CO2(-)) and its three methyl-substituted isomers (o-, m-, p-CH3C6H4CO2(-)) have been obtained using a newly developed low-temperature photoelectron spectroscopy apparatus that features an electrospray source and a cryogenically controlled ion trap. Detachment channels due to removing electrons from the carboxylate group and benzene ring pi electrons were distinctly observed. Well-resolved vibrational structures were obtained in the lower binding energy region due to the OCO bending modes, except for o-CH3C6H4CO2(-), which yielded broad spectra even at the lowest ion trap temperature (18 K). Theoretical calculations revealed a large geometry change in the OCO angles between the anion and neutral ground states, consistent with the broad ground-state bands observed for all species. A strong steric effect was observed between the carboxylate and the methyl group in o CH3C6H4CO2(-), such that the -CO2(-) group is pushed out of the plane of the benzene ring by approximately 25 degrees and its internal rotational barrier is significantly reduced. The low rotational barrier in o-CH3C6H4CO2(-), which makes it very difficult to be cooled vibrationally, and the strong coupling between the OCO bending and CO2 torsional modes yielded the broad PES spectra for this isomer. It is shown that there is no C-H...O hydrogen bond in o-CH3C6H4CO2(-), and the interaction between the carboxylate and methyl groups in this anion is found to be repulsive in nature. PMID- 16354027 TI - Stable and long-lived trianions in the gas phase. AB - Stable doubly charged anions have become well known over the past decade, but the knowledge about higher charged molecules is still sparse. In this article, we discuss the current status of trianions. The different species, both from experimental and theoretical work, are classified according to their bonding characteristics, that is, ionic, metallic, or covalent. Both stability with respect to electron autodetachment and with respect to dissociation is covered. New results on the currently smallest stable covalently bound trianion are also shown. Last, we outline future perspectives in the field of multiply charged anions. PMID- 16354028 TI - Negatively charged xanthine. I. Anions formed by canonical isomers. AB - The possibility of an excess electron binding to canonical isomers of xanthine in the gas phase was studied at the coupled-cluster level with single and double excitations using the 6-31++G** basis sets supplemented with the 4(sp)3d set of diffuse functions. It was found that xanthine should exist in the gas phase as one canonical tautomer while all the other tautomers are not likely to be detected experimentally because of their significant thermodynamic instability. On the other hand, all canonical tautomers (except one) were found to be capable of forming electronically stable anionic states of dipole-bound nature. However, the only thermodynamically stable anion (with vertical electron binding energy estimated to be 0.041 eV) based on xanthine tautomers is the one supported by the most stable neutral species. PMID- 16354029 TI - Highly compact wave functions for He-like systems. AB - Wave functions which are compact, but still quite accurate, are extremely valuable as tools for gaining understanding of quantum systems. This paper investigates the use for that purpose of functions that depend exponentially on all the interparticle distances of a few-body system, illustrated by a study of the ground electronic states of the He isoelectronic series (Z from 1 to 10). Using as few as 4 exponential basis functions, it is found that nonrelativistic energies are reproduced to within 38 microhartrees of the exact values, an error far less than for previously reported compact wave functions. Other properties are also well-represented. PMID- 16354030 TI - Derivation of quantum langevin equation from an explicit molecule-medium treatment in interaction picture. AB - A quantum mechanical form of the Langevin equation is derived from an explicit consideration of the molecule-medium interaction, as advocated by Simons in 1978, and by using two identities in the interaction picture. This can be easily reduced to the classical regime, and further simplified to the macroscopic Langevin equation by considering the stochastic Langevin force autocorrelation function. One of the so-called Einstein relations appears as a byproduct. By following the methodology proposed by Simons, an exact expression for the momentum autocorrelation function is obtained. The latter can be used to calculate the zero-frequency macroscopic diffusion coefficient that is observed to satisfy the second Einstein relation. The formalism described above gives rise to the possibility of explicitly computing the transport characteristics such as friction constant and diffusion coefficient from the corresponding quantum statistical mechanical expressions. A discussion on the Langevin equation becomes complete only when the corresponding Fokker-Planck equation is obtained. Therefore, the probability of the evolution of states with a particular absolute magnitude of linear momentum from those of another momentum eigenvalue is quantum mechanically defined. This probability appears as a special average value of a projection operator and as a special projection operator correlation function. A classical identity is introduced that is shown to be valid also for the quantum mechanically defined probability function. By using this identity, the so-called Fokker-Planck equation for the evolution probability is easily established. PMID- 16354031 TI - Length and substituent-scrambling energies of parent and halogen-substituted conjugated polyynes. AB - Conjugated polyynes are a class of species of diverse and increasing interest. Length-scrambling and substituent scrambling reaction energies were examined using ab initio quantum chemistry calculations to investigate issues concerning the energetic effects of the molecular ends (substituent communication). Computations were performed for the parent, monohalogenated, and dihalogenated (F, Cl, Br, I) polyynes of up to 60 carbon atoms. A study of resonance effects using natural resonance theory and bond lengths demonstrates lone-pair-donating effects that increase in the series F < Cl < Br < I, but run counter to the halogen inductive effects which decrease in this series and dominate energetic effects. PMID- 16354032 TI - Analysis of local anharmonicity using Gaussian model potentials and cartesian oscillator basis sets: example, HCN. AB - This paper examines local anharmonic vibrations in molecules using an analysis that starts with an ab initio potential energy surface, fits a model potential constructed of Gaussian basis functions, and proceeds to a quantum mechanical analysis of the anharmonic modes using Cartesian harmonic oscillator basis functions in a variational calculation. The objective of this work is to suggest methods, with origins in nuclear and molecular (electronic) quantum mechanics, that should be useful for the accurate analysis of the local anharmonic motions of hydrogen, and perhaps other atoms or small molecular fragments, residing in molecularly complicated but otherwise harmonic environments. PMID- 16354033 TI - Singlet-triplet splittings and ground- and excited-state electron affinities of selected cyanosilylenes, XSiCN (X = H, F, Cl, CH3, SiH3, CN). AB - Several cyanosilylenes, XSiCN, (X = H, F, Cl, CH3, SiH3, CN) have been investigated using the RHF-ACPF and CAS(2,2)-ACPF methods in conjunction with the aug-cc-pVTZ basis sets. All silylenes are found to have singlet ground states. The ground-state electron affinities are found to be rather high, i.e., 1.832, 1.497, 1.896, 1.492, 2.235, and 2.631 eV for HSiCN, FSiCN, ClSiCN, H3CSiCN, H3SiSiCN, and Si(CN)2, respectively. The existence of bound excited negative ion states has been discovered for the first time within these silylenes. All these bound excited anion states belong to the totally symmetric irreducible representations and can be characterized as dipole-bound negative ion states. All triplet excited states have even larger dipole moments than the singlet states and are, therefore, "dressed" by dipole-bound negative ion states, which correspond to Feshbach resonances. PMID- 16354034 TI - Solvent effects on interfacial electron transfer from Ru(4,4'-dicarboxylic acid 2,2'-bipyridine)2(NCS)2 to nanoparticulate TiO2: spectroscopy and solar photoconversion. AB - Resonance Raman spectra are reported for Ru(4,4'-dicarboxylic acid-2,2' bipyridine)2(NCS)2 (commonly called "N3") in ethanol solution and adsorbed on nanoparticulate colloidal TiO2 in ethanol (EtOH) and in acetonitrile (ACN), at wavelengths within the visible absorption band of the dye. Raman cross sections of free N3 in EtOH are found to be similar to those of N3 adsorbed on colloidal TiO2 in EtOH, and are generally lower than those of N3 on TiO2 in ACN. Strong electronic coupling mediated by surface states results in red-shifted absorption spectra and enhanced Raman signals for N3 adsorbed on nanocolloidal TiO2 in ACN compared to EtOH. In contrast, the absorption spectrum of N3 on nanocrystalline TiO2 in contact with solvent is similar for ACN and EtOH. Wavelength-dependent depolarization ratios for N3 Raman bands of both free and adsorbed N3 reveal resonance enhancement via two or more excited electronic states. Luminescence spectra of N3 adsorbed on nanocrystalline films of TiO2 and ZrO2 in contact with solvent reveal that the quantum yield of electron injection phi(ET) into TiO2 decreases in the order ACN > EtOH > DMSO. Dye-sensitized solar cells were fabricated with N3 adsorbed on nanocrystalline films of TiO2 in contact with ACN, EtOH, and DMSO solutions containing LiI/LiI3 electrolyte. Photoconversion efficiencies eta were found to be 2.6% in ACN, 1.3% in DMSO, and 0.84% in EtOH. Higher short circuit currents are found in cells using ACN, while the maximum voltage is found to be largest in DMSO. It is concluded that the increased photocurrent and quantum yield of interfacial electron transfer in acetonitrile as compared to ethanol and DMSO is primarily the result of faster electron injection of N3 when adsorbed on TiO2 in the presence of ACN as opposed to EtOH or DMSO. PMID- 16354035 TI - Moment analysis method as applied to the 2S --> 2P transition in cryogenic alkali metal/rare gas matrices. AB - The moment analysis method (MA) has been tested for the case of 2S --> 2P ([core]ns1 --> [core]np1) transitions of alkali metal atoms (M) doped into cryogenic rare gas (Rg) matrices using theoretically validated simulations. Theoretical/computational M/Rg system models are constructed with precisely defined parameters that closely mimic known M/Rg systems. Monte Carlo (MC) techniques are then employed to generate simulated absorption and magnetic circular dichroism (MCD) spectra of the 2S --> 2P M/Rg transition to which the MA method can be applied with the goal of seeing how effective the MA method is in re-extracting the M/Rg system parameters from these known simulated systems. The MA method is summarized in general, and an assessment is made of the use of the MA method in the rigid shift approximation typically used to evaluate M/Rg systems. The MC-MCD simulation technique is summarized, and validating evidence is presented. The simulation results and the assumptions used in applying MA to M/Rg systems are evaluated. The simulation results on Na/Ar demonstrate that the MA method does successfully re-extract the 2P spin-orbit coupling constant and Lande g-factor values initially used to build the simulations. However, assigning physical significance to the cubic and noncubic Jahn-Teller (JT) vibrational mode parameters in cryogenic M/Rg systems is not supported. PMID- 16354036 TI - Reappraisal of the role of size-extensive normalization for multireference coupled cluster (MRCC) theory using general model space: a valence universal MRCC approach. AB - We present a brief description of a valence-universal multireference coupled cluster (VU-MRCC) theory that can handle completely general incomplete model spaces, remaining close to the intermediate normalization (IN) condition for omega as much as possible without violating extensivity and without the use of a post facto correction. In this formalism, the connectedness of the cluster operators as well as effective Hamiltonian and hence the extensivity of the corresponding roots is achieved by invoking appropriate decoupling conditions on the special type of wave operator omega = {exp(S + X(cl))} satisfying the Bloch equations in the Fock-space S in an excitation operator and X is a closed operator (denoted by cl). This special type of wave-operator leads to a unique partition of the excitations from the model space into those generated by the cluster operators (open and quasi-open) and those generated by the effective Hamiltonian (closed). In this formulation, for every X(cl), there is a counterterm from {exp(S)}(cl) canceling each other. This leads to a connected expressions for cluster amplitudes, using the constraint omega(cl) = 1. The new form of the effective Hamiltonian preserves the extensivity of the target energies. Our analysis implies that IN for omega is a valid size-extensive normalization for certain special IMS such as the quasi-complete model space and the isolated incomplete model space. PMID- 16354037 TI - Mechanism of the aminolysis of methyl benzoate: a computational study. AB - Density functional and ab initio methods were applied in examining the possible mechanistic pathways for the reaction of methyl benzoate with ammonia. Transition state structures and energies were determined for concerted and neutral stepwise mechanisms. The theoretical results show that the two possible pathways have similar activation energies. The general base catalysis of the process was also examined. The predictions reveal that the catalytic process results in considerable energy savings and the most favorable pathway of the reaction is through a general-base-catalyzed neutral stepwise mechanism. The structure and transition vectors of the transition states indicate that the catalytic role of ammonia is realized by facilitating the proton-transfer processes. Comparison of the energetics of the aminolysis for methyl benzoate and methyl formate shows the more favorable process to be that for the aliphatic ester. The differing reactivity of the two esters is explained in terms of the electrostatic potential values at the atoms of the ester functionality. PMID- 16354038 TI - Dynamics of charge-transfer-to-solvent precursor states in I- (water)n (n = 3-10) clusters studied with photoelectron imaging. AB - The dynamics of charge-transfer-to-solvent states are studied in I- (H2O)(n=3-10) clusters and their deuterated counterparts using time-resolved photoelectron imaging. The photoelectron spectra for clusters with n > or = 5 reveal multiple time scales for dynamics after their electronic excitation. An increase in the vertical detachment energy (VDE) by several hundred millielectronvolts on a time scale of approximately 1 ps is attributed to stabilization of the excess electron, primarily through rearrangement of the solvent molecules, but a contribution to this stabilization from motion of the I atom cannot be ruled out. The VDE drops by approximately 50 meV on a time scale of tens of picoseconds; this is attributed to loss of the neutral iodine atom. Finally, the pump-probe signal decays with a time constant of 60 ps-3 ns, increasing with cluster size. This decay is commensurate with the growth of very slow electrons and is attributed to autodetachment. Smaller clusters (n = 3, 4) display simpler dynamics. Anisotropy parameters are reported for clusters n = 4-9. PMID- 16354039 TI - Interactions in open-shell clusters: ab initio study of pre-reactive complex O(3P) + HCl. AB - Van der Waals interactions between the ground-state triplet O(3P) atom and the closed-shell HCl molecule are investigated in the pre-reactive region. Three adiabatic (two of A'' symmetry and one of A' symmetry) and four non-relativistic diabatic potential energy surfaces are obtained by combining a restricted open shell coupled cluster approach with the multireference configuration interaction method. The lower A'' adiabatic potential surface has a single minimum (D(e) = 589 cm(-1)) for a linear O...HCl configuration. The upper A'' potential has a weak (D(e) = 65 cm(-1)) minimum for a linear HCl...O configuration. The A' adiabatic potential has a weak (124 cm(-1)) T-shaped minimum. Adiabatic potentials intersect once in the O...HCl linear configuration and twice in the linear HCl...O geometry. The role of electrostatic interactions in shaping these potentials is discussed. The effects of spin-orbit coupling on this interaction are also investigated assuming a constant value of the SO parameter. PMID- 16354040 TI - Stabilization of very rare tautomers of 1-methylcytosine by an excess electron. AB - We characterized valence anionic states of 1-methylcytosine using various electronic structure methods. We found that the most stable valence anion is related to neither the canonical amino-oxo nor a rare imino-oxo tautomer, in which a proton is transferred from the N4 to N3 atom. Instead, it is related to an imino-oxo tautomer, in which the C5 atom is protonated. This anion is characterized by an electron vertical detachment energy (VDE) of 2.12 eV and it is more stable than the anion based on the canonical tautomer by 1.0 kcal/mol. The latter is characterized by a VDE of 0.31 eV. Another unusual low-lying imino oxo tautomer with a VDE of 3.60 eV has the C6 atom protonated and is 3.6 kcal/mol less stable than the anion of the canonical tautomer. All these anionic states are adiabatically unbound with respect to the canonical amino-oxo neutral, with the instability of 5.8 kcal/mol for the most stable valence anion. The mechanism of formation of anionic tautomers with carbon atoms protonated may involve intermolecular proton transfer or dissociative electron attachment to the canonical neutral tautomer followed by a barrier-free attachment of a hydrogen atom to the C5 or C6 atom. The six-member ring structure of anionic tautomers with carbon atoms protonated is unstable upon an excess electron detachment. Indeed the neutral systems collapse without a barrier to a linear or a bicyclo structure, which might be viewed as lesions to DNA or RNA. Within the PCM hydration model, the anions become adiabatically bound with respect to the corresponding neutrals, and the two most stable tautomers have a carbon atom protonated. PMID- 16354041 TI - Photoelectron spectra and ion chemistry of imidazolide. AB - The 351.1 nm photoelectron spectrum of imidazolide anion has been measured. The electron affinity (EA) of the imidazolyl radical is determined to be 2.613 +/- 0.006 eV. Vibrational frequencies of 955 +/- 15 and 1365 +/- 20 cm(-1) are observed in the spectrum of the (2)B1 ground state of the imidazolyl radical. The main features in the spectrum are well-reproduced by Franck-Condon simulation based on the optimized geometries and the normal modes obtained at the B3LYP/6 311++G(d,p) level of density functional theory. The two vibrational frequencies are assigned to totally symmetric modes with C-C and N-C stretching motions. Overtone peaks of an in-plane nontotally symmetric mode are observed in the spectrum and attributed to Fermi resonance. Also observed is the photoelectron spectrum of the anion formed by deprotonation of imidazole at the C5 position. The EA of the corresponding radical, 5-imidazolyl, is 1.992 +/- 0.010 eV. The gas phase acidity of imidazole has been determined using a flowing afterglow-selected ion tube; delta(acid)G298 = 342.6 +/- 0.4 and delta(acid)H298 = 349.7 +/- 0.5 kcal mol(-1). From the EA of imidazolyl radical and gas phase acidity of imidazole, the bond dissociation energy for the N-H bond in imidazole is determined to be 95.1 +/- 0.5 kcal mol(-1). These thermodynamic parameters for imidazole and imidazolyl radical are compared with those for pyrrole and pyrrolyl radical, and the effects of the additional N atom in the five-membered ring are discussed. PMID- 16354042 TI - Molecular dynamics of complex gas-phase reactive systems by time-dependent groups. AB - A novel way of assembling the total potential for performing molecular dynamical studies of complex gas-phase reactive chemical systems is introduced. The method breaks the calculation of the total potential and gradients of the potential into time-dependent groups that are governed by spatial cutoffs. These groups evolve during the course of the simulation and their number may increase or diminish as the dynamics of the system determine. In an effort to extend the simulation time of these complex reactive processes and to use high levels of theory when necessary, multiple levels of theory may be used over the groups for the calculation of both the intragroup and intergroup interactions. Representative simulations are performed to illustrate the method and a computationally facile method of obtaining the groups of a simulation are also discussed. PMID- 16354043 TI - Electronic structure differences in ZrO2 vs HfO2. AB - Although ZrO2 and HfO2 are, for the most part, quite similar chemically, subtle differences in their electronic structures appear to be responsible for differing MO2/Si (M = Zr, Hf) interface stabilities. To shed light on the electronic structure differences between ZrO2 and HfO2, we have conducted joint experimental and theoretical studies. Because molecular electron affinities are a sensitive probe of electronic structure, we have measured them by conducting photoelectron spectroscopic experiments on ZrO2(-) and HfO2(-). The adiabatic electron affinity of HfO2 was determined to be 2.14 +/- 0.03 eV, and that of ZrO2 was determined to be 1.64 +/- 0.03 eV. Concurrently, advanced electronic structure calculations were conducted to determine electron affinities, vibrational frequencies, and geometries of these systems. The calculated CCSD(T) electron affinities of HfO2 and ZrO2 were found to be 2.05 and 1.62 eV, respectively. The molecular results confirm earlier predictions from solid state calculations that HfO2 is more ionic than ZrO2. The excess electron in MO2(-) occupies an sd-type hybrid orbital localized on the M atom (M = Zr, Hf). The structural parameters of ZrO2 and HfO2 and their vibrational frequencies were found to be very similar. Upon the excess electron attachment, the M-O bond length increases by ca. 0.04 A, the OMO angle increases by 2-4 degrees, and frequencies of all vibrational modes become smaller, with the stretching modes being shifted by 30-50 cm(-1) and the bending mode by 15-25 cm(-1). Together, these studies unveil significant differences in the electronic structures of ZrO2 and HfO2 but not in their structural or vibrational characteristics. PMID- 16354044 TI - Infrared spectrum and structural assignment of the water trimer anion. AB - The bending vibrational spectrum of the perdeutero isotopomer of the water trimer anion has been measured and compared with spectra calculated using the MP2, CCSD, and Becke3LYP electronic structure methods. Due to its low electron binding energy (approximately 150 meV), only the OD bending region of the IR spectrum of (D2O)3(-) is accessible experimentally, with electron ejection dominating at higher photon energies. The calculated spectrum of the isomer having three water molecules arranged in a chain agrees best with the experimental spectrum. In the chain isomer, the excess electron is bound to the terminal water monomer with two dangling OH groups. This is consistent with the electron binding mechanism established previously for the (H2O)n(-) (n = 2, 4-6) anions. PMID- 16354045 TI - Quantum Drude oscillator model for describing the interaction of excess electrons with water clusters: an application to (H2O)13(-). AB - Cluster anions for which the excess electron occupies an extended nonvalence orbital can be described by use of a model Hamiltonian employing quantum Drude oscillators to represent the polarizable charge distributions of the monomers. In this work, a Drude model for water cluster anions is described and used to investigate the (H2O)13(-) cluster. Several low-energy isomers are characterized, and the finite-temperature properties of the cluster are investigated by means of parallel tempering Monte Carlo simulations. Two structural motifs, one with double-acceptor water monomers and the other with four-membered rings of double acceptor single-donor monomers with four free OH groups pointed in the same direction, are found to lead to large (approximately > eV) electron binding energies. The distributions of the computed vertical detachment energies qualitatively reproduce the experimentally measured photoelectron spectrum, and our simulations indicate that both of the main peaks in the measured spectrum derive from several isomers. PMID- 16354047 TI - Dissociation of benzene dication [C6H6]2+: exploring the potential energy surface. AB - The singlet potential energy surface for the dissociation of benzene dication has been explored, and its three major dissociation channels have been studied: C6H6(2+) --> C3H3(+) + C3H3(+), C4H3(+) + C2H3(+), and C5H3(+) + CH3(+). The calculated energetics suggest that the products will be formed with considerable translational energy because of the Coulomb repulsion between the charged fragments. The calculated energy release in the three channels shows a qualitative agreement with the experimentally observed kinetic energy release. The formation of certain intermediates is found to be common to the three dissociation channels. PMID- 16354046 TI - Cation-pi interactions with a model for the side chain of tryptophan: structures and absolute binding energies of alkali metal cation-indole complexes. AB - Threshold collision-induced dissociation techniques are employed to determine bond dissociation energies (BDEs) of mono- and bis-complexes of alkali metal cations, Li+, Na+, K+, Rb+, and Cs+, with indole, C8H7N. The primary and lowest energy dissociation pathway in all cases is endothermic loss of an intact indole ligand. Sequential loss of a second indole ligand is observed at elevated energies for the bis-complexes. Density functional theory calculations at the B3LYP/6-31G level of theory are used to determine the structures, vibrational frequencies, and rotational constants of these complexes. Theoretical BDEs are determined from single point energy calculations at the MP2(full)/6-311+G(2d,2p) level using the B3LYP/6-31G* geometries. The agreement between theory and experiment is very good for all complexes except Li+ (C8H7N), where theory underestimates the strength of the binding. The trends in the BDEs of these alkali metal cation-indole complexes are compared with the analogous benzene and naphthalene complexes to examine the influence of the extended pi network and heteroatom on the strength of the cation-pi interaction. The Na+ and K+ binding affinities of benzene, phenol, and indole are also compared to those of the aromatic amino acids, phenylalanine, tyrosine, and tryptophan to elucidate the factors that contribute to the binding in complexes to the aromatic amino acids. The nature of the binding and trends in the BDEs of cation-pi complexes between alkali metal cations and benzene, phenol, and indole are examined to help understand nature's preference for engaging tryptophan over phenylalanine and tyrosine in cation-pi interactions in biological systems. PMID- 16354048 TI - MX3(-) superhalogens (M = Be, Mg, Ca; X = Cl, Br): a photoelectron spectroscopic and ab initio theoretical study. AB - Gas-phase alkaline earth halide anions, MgX3(-) and CaX3(-) (X = Cl, Br), were produced using electrospray and investigated using photoelectron spectroscopy at 157 nm. Extremely high electron binding energies were observed for all species and their first vertical detachment energies were measured as 6.60 +/- 0.04 eV for MgCl3(-), 6.00 +/- 0.04 eV for MgBr3(-), 6.62 +/- 0.04 eV for CaCl3(-), and 6.10 +/- 0.04 eV for CaBr3(-). The high electron binding energies indicate these are very stable anions and they belong to a class of anions, called superhalogens. Theoretical calculations at several levels of theory were carried out on these species, as well as the analogous BeX3(-). Vertical detachment energy spectra were predicted to compare with the experimental observations, and good agreement was obtained for all species. The first adiabatic detachment energies were found to be substantially lower (by about 1 eV) than the corresponding vertical detachment energies for all the MX3(-) species, indicating extremely large geometry changes between MX3(-) and MX3. We found that all the MX3(-) anions possess D3h ((1)A1') structures and are extremely stable against dissociation into MX2 and X-. The corresponding neutral species MX3, however, were found to be only weakly bound with respect to dissociation toward MX2 + X. The global minimum structures of all the MX3 neutrals were found to be C2v ((2)B2), which can be described as (X2(-))(MX+) charge-transfer complexes, whereas the MX2...X (C2v, (2)B1) van der Waals complexes were shown to be low lying isomers. PMID- 16354049 TI - Complexation of the carbonate, nitrate, and acetate anions with the uranyl dication: density functional studies with relativistic effective core potentials. AB - The structures and vibrational frequencies of uranyl carbonates, [UO2(CO3)n](2 2n) and [(UO2)3(CO3)6]6-, uranyl nitrates, [UO2(NO3)n](2-n), and uranyl acetates, [UO2(CH3COO)n](2-n) (n = 1,2,3) have been calculated by using local density functional theory (LDFT). Only bidentate ligand coordination modes to the uranyl dication have been modeled. The calculated structures and frequencies are compared to available experimental data, including IR, Raman, X-ray diffraction, and EXAFS solution and crystal structure data. The energetics of ligand binding have been calculated using the B3LYP hybrid functional. In general, the structural and vibrational results at the LDFT level are in good agreement with experimental results and provide realistic pictures of solution phase and solid state behavior. For the [UO2(CO3)3]6- anion, calculations suggest that complexity in the CO3(2-) stretching signature upon complexation is due to the formation of C=O and C-O domains, the latter of which can split by as much as 300 cm(-1). Assessment of the binding energies indicate that the [UO2(CO3)2]2- anion is more stable than the [UO2(CO3)3]4- anion due to the accumulation of excess charge, whereas the tri-ligand species are the most stable in the nitrate and acetate anions. PMID- 16354050 TI - Quasiresonance: switching internal energy transfer on and off. AB - Quasiresonance involves a slow "external" switching on and off of an interaction between internal degrees of freedom described by action-angle variables having approximate resonances. The resonances or near-resonances spawn slow coordinates that fail to be adiabatic, but the remaining coordinates may be fast enough to have conserved actions. The interaction either can be imposed externally as a time dependent coupling or can arise autonomously due to interactions with other degrees of freedom. A resonance transformation into slow and fast angles reveals the action corresponding to the fast angle is adiabatic and conserved to very high accuracy. This paper extends our work on quasiresonance to new systems and regimes, including the He-H2 system, collisions with a periodic lattice, perturbative interactions, and discussion of quasiresonance in higher dimensional systems. PMID- 16354051 TI - Intermolecular potential of the O2-O2 dimer. An ab initio study and comparison with experiment. AB - Accurate intermolecular potentials for the lowest three multiplet states of O2-O2 dimer have been produced on the basis of ab initio calculations. The quintet potential was taken from previous highly correlated CCSD(T) calculations. In this work, we perform MRCI calculations, with large basis sets including bond functions, of the singlet and triplet states, which are of multireference character. As expected the size inconsistency and lack of higher order excitations limit the accuracy of the MRCI potentials specifically in describing the long range interactions. We show that the Heisenberg Hamiltonian provides an accurate representation of the exchange interactions in this system and this enables us to combine the accurate CCSD(T) potentials with the MRCI spin-exchange parameter to obtain accurate singlet and triplet potentials. The reliability of these potentials is tested by computing integral cross sections and comparing them with the detailed experimental study of the Perugia group, with excellent results. More interestingly, comparison with the experimentally derived potential shows important discrepancies for some angular orientations including that corresponding with the global minima, indicating the need for further work, both theoretical and experimental, to clarify their origin. PMID- 16354052 TI - Ab initio electron propagator calculations on the ionization energies of free base porphine, magnesium porphyrin, and zinc porphyrin. AB - Ab initio electron propagator methods are applied to the prediction, assignment, and interpretation of the valence photoelectron spectra of free base porphine and of magnesium and zinc porphyrins. Tests of various approximate self-energies, including the partial third (P3), the outer valence Green's function, and the nondiagonal, renormalized second-order (NR2) methods are performed. Basis set effects and reduced active orbital spaces are examined as well. The P3 method and the one-electron picture of ionization that accompanies it are validated for the first two cationic states and for states with sigma holes that are localized in nitrogen, lone pair regions. In the remaining pi-hole states, there is significant shake-up character and NR2 results provide useful diagnostics of correlation effects. PMID- 16354053 TI - p-Difluorobenzene-argon ground state intermolecular potential energy surface. AB - The ground state intermolecular potential energy surface for the p difluorobenzene-Ar van der Waals complex is evaluated using the coupled cluster singles and doubles including connected triple excitations [CCSD(T)] model and the augmented correlation consistent polarized valence double-zeta basis set extended with a set of 3s3p2d1f1g midbond functions. The surface minima are characterized by the Ar atom located above and below the difluorobenzene center of mass at a distance of 3.5290 A. The corresponding binding energy is -398.856 cm(-1). The surface is used in the evaluation of the intermolecular level structure of the complex. The results clearly improve previously available data and show the importance of using a good correlation method and basis set when dealing with van der Waals complexes. PMID- 16354054 TI - An efficient and accurate implementation of centroid molecular dynamics using a Gaussian approximation. AB - An approximate method for Centroid Molecular Dynamics (CMD) is presented which uses a Gaussian approximation. The resulting method, called Gaussian CMD (GCMD), is 100-1000 times faster than CMD because it replaces explicit path-integral sampling, which is the most time-consuming part of CMD, with a Gaussian averaging, which can be done analytically. Several methods for computing the Gaussian width parameter in the GCMD approach are also presented. This new method is shown to give satisfactory results for the position correlation function in simple one-dimensional systems when CMD itself is consistent with the exact result. The GCMD and CMD results are also compared for the case of 1-dimensional systems coupled to harmonic baths, with good success. GCMD is further compared to CMD with good success for liquid para-hydrogen at two different temperatures, 14 K and 25 K, and for ortho-deuterium at 20.7 K. PMID- 16354055 TI - Quadratic response functions in a second-order polarization propagator framework. AB - The linear and quadratic response functions have been derived for an exact state, based on an exponential parametrization of the time evolution consisting of products of exponentials for orbital rotations and for higher-order excitations. Truncating the linear response function such that the response function itself and its pole structure is correct to second order in Moller-Plesset perturbation theory, we arrive at the second-order polarization propagator approximation (SOPPA). Previous derivations of SOPPA have used the superoperator formalism, making the extension of SOPPA to quadratic and higher order response functions difficult. The derivation of the quadratic response function is described in detail, allowing molecular properties such as hyperpolarizabilities, two-photon cross sections, and excited-state properties to be calculated using the SOPPA model. PMID- 16354056 TI - A general nonlinear expansion form for electronic wave functions. AB - A new expansion form is presented for electronic wave functions. The wave function is a linear combination of product basis functions, and each product basis function in turn is formally equivalent to a linear combination of configuration state functions that comprise an underlying linear expansion space. The expansion coefficients that define the basis functions are nonlinear functions of a smaller number of variables. The expansion form is appropriate for both ground and excited states and to both closed and open shell molecules. The method is formulated in terms of spin-eigenfunctions using the graphical unitary group approach (GUGA), and consequently it does not suffer from spin contamination. PMID- 16354057 TI - Meso-structure formation for enhanced organic photovoltaic cells. AB - [reaction: see text] Formation of a controlled fullerene mesophase within an organic host system has enabled us to create high-power conversion efficiency photovoltaics. This mesophase is formed using thermal gradients that provide a fluidic mobility of the fullerenes allowing for greater dispersion of nanocrystalline 1-(3-methoxycarbonyl)propyl-1-phenyl-(6,6)C61 (PCBM) within regioregular poly(3-hexylthiophene) (P3HT). From this reorganization of the component materials in the matrix the overall efficiency of the system jumps dramatically from the roughly 2.4% to 5.2%. PMID- 16354058 TI - A fast-moving [2]rotaxane whose stoppers are remote from the copper complex core. AB - [reaction: see text] A new copper-complexed rotaxane is described. It consists of a two-coordination site ring threaded by a sterically non-hindering 2,2' bipyridine derivative. An electrochemical signal (oxidation or reduction of the copper center, Cu(I) or Cu(II)) induces rearrangement of the system. By using long and flexible linkers between the stoppers and the central complex, ligand exchange is fast, which leads to short response times (on the millisecond time scale and even below). PMID- 16354059 TI - Quantum mechanical calculations of conformationally relevant 1H and 13C NMR chemical shifts of calixarene systems. AB - [graphs: see text] QM GIAO calculations of 13C and 1H chemical shift values of the ArCH2Ar group have been performed, using the hybrid DFT functional MPW1PW91 and the 6-31G(d,p) basis set, on some representative calixarenes and on a series of simplified calixarene models allowing derivation of chemical shift surfaces versus phi and chi dihedral angles. A good reproduction of experimental data was obtained. The applicability of chemical shift surfaces in the study of calixarene conformational features is illustrated. PMID- 16354060 TI - Star-shaped hexa-peri-hexabenzocoronene "heptamer": synthesis and self-assembly. AB - [structure: see text] A star-shaped hexa-peri-hexabenzocoronene "heptamer" was prepared, which showed a strong tendency to aggregate in solution and in the bulk states. Higher order was found in a high-temperature, columnar liquid crystalline phase due to the higher mobility of the molecules in comparison with the low temperature solid. In addition, physical gel formation was observed due to the presence of covalent intercolumnar interactions. PMID- 16354061 TI - Synthesis of a novel cyclic donor-acceptor conjugate for selective recognition of ATP. AB - [reaction and structures: see text] Novel cyclophane 1 was synthesized, and its interactions with phosphate, adenosine, AMP, ADP, and ATP have been investigated. With addition of ATP, significant decrease in absorbance of 1 was observed, whereas other guest molecules showed negligible effect. The complex between 1 and ATP was confirmed through cyclic voltammetry and 1H NMR. The uniqueness of the system is that it complexes selectively with ATP in a cavity and involves synergistic effects of both electrostatic and pi-pi stacking interactions. PMID- 16354062 TI - Synthesis of a versatile (S)-3-(hydroxymethyl)butane-1,2,4-triol building block and its application for the stereoselective synthesis of N-homoceramides. AB - [structures: see text] A versatile (S)-3-(hydroxymethyl)butane-1,2,4-triol building block has been synthesized starting from D-isoascorbic acid, a common food preservative. The key transformation in this approach was the introduction of branching through a high yield and fully regioselective epoxide opening. This flexible synthon has been elaborated to a new class of (dihydro-)N homo(phyto)ceramides. PMID- 16354063 TI - A Diels-Alder approach to the stereoselective synthesis of 2,3,5,6-tetra- and 2,3,4,5,6-pentasubstituted piperidines. AB - [structures: see text] A stereoselective synthesis of 2,3,5,6-tetra- and 2,3,4,5,6-pentasubstituted piperidines was achieved from oxidative cleavage of 2 aza-bicyclo[2.2.2]octene Diels-Alder adducts derived from N-protected 2-methyl 1,2-dihydropyridine. A chiral auxiliary mediated asymmetric synthesis of the pentasubstituted piperidine is also demonstrated. This methodology incorporates orthogonal protecting groups, thus providing a piperidine scaffold with easily modified points of diversity. PMID- 16354064 TI - Tropos or atropos nature of rhodium complexes bearing a tetrakis(phosphanyl)terphenyl ligand: highly enantioselective catalysis of ene type cyclization. AB - [reaction and structure: see text] Not only axial but also helical chirality of tropos TETRAPHOS-Rh complexes can be controlled by chiral diamines. The flexibility of the TETRAPHOS-Rh complex is increased by association of DABN. In contrast, the diamine-free complex is chirally more stable than the BIPHEP counterpart. The higher levels of enantioselectivity in ene-type cyclization of 1,6-enynes can thus be achieved even at room temperature by the diamine-free TETRAPHOS-Rh complex. PMID- 16354065 TI - Synthesis of functionalized hydroquinones via [Cp*RuCl2]2-catalyzed cocyclization of alkynes, alpha,beta-unsaturated carbonyl compounds, and carbon monoxide. AB - [reaction: see text] Catalytic [2 + 2 + 1 + 1] cocyclization reaction of an alkyne, an alkene, and two molecules of carbon monoxide, leading to functionalized hydroquinones, was studied. Using [Cp*RuCl2]2 as a catalyst, we found that a variety of electron-deficient alkenes, such as alpha,beta unsaturated ketones, esters, amides, and nitriles, can be employed as an alkene coupling partner to give the corresponding hydroquinones. PMID- 16354066 TI - Lewis acid-mediated generation of bicyclo[5.3.0]decanes and bicyclo[4.3.0]nonanes. AB - [reactions: see text] Fragmentation of the cyclobutane-containing adducts generated from intramolecular cycloadditions of cyclobutadiene with olefins provides rapid entry into bicyclo[5.3.0]decane and bicyclo[4.3.0]nonane ring systems. Whereas earlier studies featured thermal methods to achieve the desired rearrangements, a mild, Lewis acid-mediated fragmentation has been identified for substrates with appropriate functionality adjacent to the strained ring system. The substrate scope and stereochemical outcome of the acid-mediated fragmentation are complementary to the thermal ring expansions, particularly in the case of the bicyclo[5.3.0]decanes. PMID- 16354067 TI - Telluronium salts mediated aziridination of chiral N-tert-butylsulfinylimines: highly stereoselective synthesis of optically active vinylaziridines. AB - [reaction: see text] Optically active cis-2-substituted vinylaziridines are synthesized by the reaction of N-tert-butylsulfinylimines with telluronium ylides with excellent diastereoselectivity in good to excellent yields. PMID- 16354068 TI - A new entry to the synthesis of 2,3-disubstituted indoles. AB - [reaction: see text] The Stille coupling of N-acyl-2-iodoanilines with the 1 (tributylstannyl)-1-substituted allenes affected the successive one-step formation of the 2-methyl-3-substituted indoles. Alternatively, the other type of 2-alkyl-3-substituted indoles could be synthesized in a one-pot operation, which consists of the Stille coupling reaction with the 1-(tributylstannyl)-1,3 disubstituted allenes, followed by TBAF treatment. This procedure could be applied to the synthesis of indomethacin. PMID- 16354069 TI - Addition of a chiral copper reagent derived from propargylic oxazolidininone to aldehydes. AB - [reaction: see text] The copper reagent arising from an optically pure propargylic oxazolidinone was found to react regio- and diastereoselectively with aldehydes, leading, in a one-pot procedure, to the anti homopropargylic amino alcohols derivatives in good yields. PMID- 16354070 TI - Aryl iodide mediated aziridination of alkenes. AB - [reaction: see text] Aryl iodide mediated aziridination of a variety of alkenes with N-aminophthalimide under mild conditions (m-CPBA, K2CO3, CH2Cl2, 25 degrees C) was achieved in moderate to good yields (up to 94%). By recovering the aryl iodide, a recyclable system is developed with product yield over 79% attained for the aziridination of trans-1,2-diphenylethylene. PMID- 16354071 TI - An efficient synthesis of D-erythro- and D-threo-sphingosine from D-glucose: olefin cross-metathesis approach. AB - [reaction: see text] The D-erythro- and D-threo-sphingosine were synthesized via E-selective olefin cross-metathesis using a D-glucose-derived building block and long-chain terminal alkene. PMID- 16354072 TI - Stereoselective pinacol coupling of chiral formylferrocene using divalent samarium triflate: preparation of a new chiral bisferrocenyl oxazoline ligand and its application to asymmetric Diels-Alder reactions. AB - [reaction: see text] The pinacol coupling reaction of planar chiral ortho oxazoline-substituted formylferrocene was smoothly mediated by SmI2 or Sm(OTf)3 to give the (R,R) isomer selectivity (up to 76% diastereomeric excess). The combination of Yb(OTf)3 and the (R,R)-ferrocenyl diol was revealed to be a good catalyst for the asymmetric Diels-Alder reaction of 3-acyloxazolidinone with cyclopentadiene, and the endo adduct was produced in up to 80% enantiomeric excess. PMID- 16354073 TI - A total synthesis of tarchonanthuslactone exploiting N-pyrrole carbinols as efficient stereocontrolling elements. AB - [reaction: see text] A short stereoselective total synthesis of the polyketide natural product, tarchonanthuslactone, has been achieved. The key sequence involves the first reported catalytic enantioselective reduction of an N-acyl pyrrole and subsequent use of this stereocenter in a diastereoselective reductive cascade. This proceeded with unprecedentedly high stereocontrol and offered an elegant method of generating the desired syn stereochemistry present in the final target in one step. PMID- 16354074 TI - A theoretical study of the [4 + 4] dimerization of thioformylketene. AB - [reaction and structure: see text] A theoretical study (B3LYP and G3MP2B3) of the dimerization of thioformylketene (1) was performed. Four pathways-two [4 + 2] pathways with thioformylketene (1), one [4 + 4] pathway with 1, and one [4 + 2] pathway involving 1 and thietone (11)-were considered. Interestingly, the [4 + 4] pathway with 1 had the lowest barrier (3.8 kcal/mol). The geometry of the transition state TS14 is unusual, with the forming bonds in the plane of the ketene. This suggests that the reaction is pseudopericyclic. PMID- 16354075 TI - Highly efficient chromatographic resolution of alpha,alpha'-dihydroxybiaryls. AB - [graph: see text] Separation factors as high as 115 were observed for the chromatographic resolution of many alpha,alpha'-dihydroxybiaryls with a single chiral stationary phase made from readily available amino acid derivatives. The stationary phase works well for biphenyl-type compounds. It works extremely well for larger bis-aromatic compounds, such as binaphthyl-type compounds. PMID- 16354076 TI - Cholic-acid-based fluorescent sensor for dicarboxylates and acidic amino acids in aqueous solutions. AB - [structure: see text] The binding affinities of a cholic-acid-based fluorescent neutral receptor toward dicarboxylate anions and amino acids have been investigated in a CH3OH/H2O system (1:1, 0.01 M HEPES buffer, pH = 7.4) by fluorescence titration experiments. The synthetic host bearing four convergent functionalities strongly binds glutamate via multiple hydrogen bonds with a binding constant of (5.57 +/- 0.88) x 10(6). PMID- 16354077 TI - Monitoring of microenvironmental changes in the major and minor grooves of DNA by dan-modified oligonucleotides. AB - [graph: see text] We describe the synthesis of new environmentally sensitive fluorescence probes to elucidate DNA structures. DNA oligonucleotides containing fluorophore dan (6-(dimethylamino)-2-acylnaphthalene)-modified dC or dG were able to monitor the microenvironmental changes in both the major and minor grooves of DNA with a B- to A-DNA conformational transition and RNA hybridization. PMID- 16354078 TI - Photoinduced metalation of nonactivated C-Cl bonds with samarium diiodide: synthesis of alkenes with high (Z)-selectivity through beta-elimination reactions. AB - [reaction: see text] The photoinduced metalation of nonactivated C-Cl bonds of O acetyl chlorohydrins is promoted by samarium diiodide. As a result of this, beta elimination of O-acetyl chlorohydrins is achieved, affording the corresponding (Z)-alkenes with total or high stereoselectivity. PMID- 16354079 TI - Palladium-catalyzed asymmetric amination and imidation of 2,3-allenyl phosphates. AB - [reaction: see text] Asymmetric amination of 2,3-allenyl phosphates with nitrogen nucleophiles such as amines, hydroxylamines, and imides can be performed efficiently using a combination of zerovalent palladium complexes and SEGPHOS or MeOBIPHEP ligand, affording the corresponding optically active 1-aminated derivatives with enantiomeric excess of up to 97% ee. PMID- 16354080 TI - Synthesis of (+/-)-brazilin using IBX. AB - [reaction: see text] A short synthesis of (+/-)-brazilin is reported. This synthesis uses several interesting and underutilized transformations including a regioselective dirhodium-catalyzed aryl C-H insertion, a regioselective IBX phenol --> o-quinone oxidation, a tautomerization of an o-quinone to a p-quinone methide, and an intramolecular aryl cyclization with a p-quinone methide. PMID- 16354081 TI - Development of an anomalous Heck reaction: skeletal rearrangement of divinyl and enyne carbinols. AB - [reaction: see text] A general set of conditions that achieves the union of aryl halides and divinyl or enyne carbinols to afford tri- or tetrasubstituted olefins in good yields (up to 83%) is described. The mechanism by which this proceeds is believed to involve the intermediacy of a cyclopropanol, followed by a novel skeletal reorganization. The ability to suppress beta-hydride elimination of organopalladium intermediates appears to be critical to the success of these processes. PMID- 16354082 TI - A new method for the functionalization of [60] fullerene: an unusual 1,3-dipolar cycloaddition pathway leading to a C60 housane derivative. AB - [reaction: see text] A variant of the Huisgen 1,3-dipolar cycloaddition reaction provides a new and convenient functionalization of fullerenes. This method complements the widely used Prato and Bingel-Hirsch reactions. The derived, highly functionalized cyclopentenone and cyclopentenamine fullerene compounds upon hydrolysis are suitable for further functionalization and may serve well in the synthesis of new C60 derivatives possessing uncommon and interesting properties. PMID- 16354083 TI - Rhodium-catalyzed intramolecular [4 + 2] cycloadditions of alkynyl halides. AB - [reaction: see text] Cationic rhodium(I)-catalyzed intramolecular [4 + 2] cycloadditions of diene-tethered alkynyl halides were found to occur in good yields (70-87%). The halide moiety is compatible with the cycloaddition reactions, and no oxidative insertion to the alkynyl halide was observed. The halogen-containing cycloadducts could be transformed into a variety of products that are difficult or impossible to obtain via direct cycloaddition. PMID- 16354084 TI - Practical synthesis of triaryl- and triheteroarylmethanes by reaction of aldehydes and activated arenes promoted by gold(III) chloride. AB - [reaction: see text] Electron-rich arenes condense efficiently with various aldehydes under the influence of AuCl3, thus opening up a practical route to triarylmethanes, which have important applications. The mild conditions employed are especially noteworthy. PMID- 16354085 TI - 3-Aminopyrrolidines via ring rearrangement of 2-aminomethylazetidines. Synthesis of (-)-absouline. AB - [reaction: see text] A new entry to enantiopure 3-aminopyrrolidines was developed using a boron trifluoride-mediated rearrangement of 2-aminomethylazetidines. The method is quite general and produces rearranged products in good yield regardless of both substitution pattern and relative stereochemistry of the starting material. A concise stereocontrolled synthesis of (-)-absouline was achieved on the basis of this new method. PMID- 16354087 TI - New Hg2+ and Cu2+ selective chromo- and fluoroionophore based on a bichromophoric azine. AB - [graph: see text] A new probe, 1,4-bis(1-pyrenyl)-2,3-diaza-1,3-butadiene, selectively senses Hg2+ and Cu2+ through two different channels: the yellow-deep pink color change and the enhancement of the fluorescence with the red shift of the excimer emission, which can visually be discernible by a green fluorescence in the presence of Hg2+ and an orange fluorescence in the presence of Cu2+. PMID- 16354086 TI - Pericyclic reactions of prenylated naphthoquinones: biomimetic syntheses of mollugin and microphyllaquinone. AB - [reaction: see text] A total synthesis of the bioactive naphthohydroquinone mollugin and the related naphthoquinone dimer microphyllaquinone is described. Both syntheses exploit the propensity of prenylated quinones to undergo tautomerization/oxa 6pi-electrocyclizations. PMID- 16354088 TI - Enzymatic formation of indole-containing unnatural cyclic polyprenoids by bacterial squalene:hopene cyclase. AB - [reaction: see text] Two indole-containing substrate analogues, in which a C20 isoprene unit is connected to indole (3-(geranylgeranyl)indole and 3 (farnesyldimethylallyl)indole), were synthesized and tested for enzymatic cyclization by squalene:hopene cyclase from Alicyclobacillus acidocaldarius. Interestingly, 3-(geranylgeranyl)indole was not a substrate for the bacterial squalene cyclase, while 3-(farnesyldimethylallyl)indole was efficiently converted to a 2:1 mixture of unnatural novel products. PMID- 16354089 TI - A highly facile and efficient one-step synthesis of N6-adenosine and N6-2' deoxyadenosine derivatives. AB - [reaction: see text] A highly facile and efficient one-step synthesis of N6 adenosine and N6-2'-deoxyadenosine derivatives has been developed. Treatment of inosine or 2'-deoxyinosine, without protection of sugar hydroxyl groups, with alkyl or arylamines, in the presence of BOP and DIPEA in DMF, led to the formation of N6-adenosine and N6-2'-deoxyadenosine derivatives in good to excellent yields. Carcinogenic polyaromatic hydrocarbon (PAH) N6-2' deoxyadenosine adduct 10 and a rare DNA constituent 11 were thus synthesized directly from 2'-deoxyinosine both in 98% yield. PMID- 16354090 TI - Solid-phase synthesis of callipeltin D. Stereochemical confirmation of the unnatural amino acid AGDHE. AB - [structure: see text] The lipopeptide callipeltin D (1) was synthesized using an Fmoc-based solid-phase strategy in seven steps and 35% overall yield. The 1H NMR of synthetic 1 correlated closely with that of the natural product, confirming the configurational assignment of the novel amino acid constituent (2R,3R,4S)-4 amino-7-guanidino-2,3-dihydroxyheptanoic acid. PMID- 16354091 TI - Pd-catalyzed carbocyclization-Negishi cross-coupling cascade: a novel approach to 1alpha,25-dihydroxyvitamin D3 and analogues. AB - [reaction: see text] A mild palladium-catalyzed cascade has been used for the synthesis of the hormone 1alpha,25-dihydroxyvitamin D3 (calcitriol, 1a) and its analogues 1b and 1c. This one-pot process involves two consecutive transformations at room temperature: An initial palladium-catalyzed 6-exo cyclocarbopalladation of vinyl triflates followed by a Negishi cross-coupling reaction with an alkenyl zinc. This novel strategy opens new possibilities for the preparation of a variety of new vitamin D analogues of therapeutic potential, particularly with modifications at the triene and/or ring-A. PMID- 16354092 TI - Enantiomerically pure rhodium complexes bearing 1,5-diphenyl-1,5-cyclooctadiene as a chiral diene ligand. Their use as catalysts for asymmetric 1,4-addition of phenylzinc chloride. AB - [reaction and structures: see text] A rhodium complex coordinated with 1,5 diphenyl-1,5-cyclooctadiene (Ph-cod), [RhCl((R)-Ph-cod)]2, was obtained enantiomerically pure through optical resolution of diastereomeric isomers [Rh(Ph cod)((R)-1,1'-binaphthyl-2,2'-diamine)]BF4. The enantiomerically pure rhodium complexes showed high catalytic activity and enantioselectivity (up to 98% ee) in the asymmetric 1,4-addition of phenylzinc chloride to alpha,beta-unsaturated ketones and esters in the presence of chlorotrimethylsilane. PMID- 16354093 TI - Room-temperature Cannizzaro reaction under mild conditions facilitated by magnesium bromide ethyl etherate and triethylamine. AB - [reaction: see text] A room-temperature convenient Cannizzaro reaction prompted by magnesium bromide ethyl etherate and triethylamine is described for smooth conversion of aromatic aldehydes into their respective alcohols and carboxylic acids. The methodology is applicable to both inter- and intramolecular reactions and could be directed to obtain the carboxylic moiety in the form of an acid, an amide, or an ester compound depending on the selected reaction conditions or workup procedure. PMID- 16354094 TI - The first synthesis of a methano[60]fullerene with an electron-donating group at the methano-bridge carbon: synthesis and reaction of aminomethano[60]fullerene. AB - [reaction: see text] Aminomethano[60]fullerene was synthesized for the first time as a trifluoromethanesulfonic acid salt by applying the Curtius rearrangement of azidocarbonylmethano[60]fullerene as the key reaction. Aminomethano[60]fullerene thus obtained was found to be able to react with various acyl chlorides to afford the corresponding amides. PMID- 16354095 TI - Diastereoselective, titanium-mediated cyclization of omega-vinyl tethered imides. AB - [reaction: see text] Diastereoselective reductive coupling reactions of omega vinyl tethered cyclic imides are achieved by a preexisting stereocenter at an allylic position. Particularly noteworthy is the effective use of a 1:2 mixture of Ti(O-i-Pr)4 and n-BuLi to afford the N-acylhemiaminal products in good yields. PMID- 16354096 TI - Asymmetric aza-Mannich reactions of sulfinimines: scope and application to the total synthesis of a bromopyrrole alkaloid. AB - [reaction: see text] An asymmetric intermolecular aza variant of the Mannich reaction is reported utilizing chiral sulfinimine anions as the nucleophile and N sulfonyl aldimines as the electrophilic component. A wide range of nucleophiles and electrophiles are tolerated by the reaction conditions, delivering the condensation products in good to excellent yield with a high degree of stereocontrol. Application of this methodology to the total synthesis of a natural product is reported. PMID- 16354097 TI - Radical reaction of sodium hypophosphite with terminal alkynes: synthesis of 1,1 bis-H-phosphinates. AB - [reaction: see text] The room-temperature radical addition of sodium hypophosphite to terminal alkynes produces the previously unknown 1-alkyl-1,1-bis H-phosphinates in moderate yield. The reaction is initiated by R3B and air and proceeds under mild conditions in an open container. The bissodium salts precipitate spontaneously from the reaction mixtures, thus providing a simple purification procedure and the opportunity for multigram synthesis. The 1,1-bis-H phosphinate products are novel precursors of the biologically important 1,1 bisphosphonates. PMID- 16354098 TI - Asymmetric [2,3]-Wittig rearrangement induced by a chiral carbanion whose chirality was transferred from an epoxide. AB - [reaction: see text] The enantioselective [2,3]-Wittig rearrangement of 1 allyloxy-1-(naphthalen-2-yl)-4-siloxy-2,4-pentadienyl anion, derived from optically enriched 4,5-epoxy-1-(naphthalen-2-yl)-5-silyl-2-pentenyl allyl ether via a base-induced ring opening of the epoxide followed by Brook rearrangement, has been studied. The chirality of the epoxide was transferred to the alcohols in up to 97% ee, depending on the solvent used. The best result was obtained in 1,4 dioxane at a temperature above room temperature. PMID- 16354099 TI - Stereoselective and efficient synthesis of (3R,3aS,6aR)-hexahydrofuro[2,3-b]furan 3-ol. AB - [reaction: see text] Two short and efficient synthesis routes have been developed for bis-THF-alcohol 2, a key building block of the investigational HIV protease inhibitor TMC114 (1). Using S-2,3-O-isopropylideneglyceraldehyde (4) as the source of chirality, both routes are based on a diastereoselective Michael addition of nitromethane to give predominantly the syn congeners 6 followed by a Nef oxidation and cyclization to afford lactone acetals 8, which are reduced and cyclized to give 2. PMID- 16354100 TI - Synthesis of highly functionalized bicyclo[m.n.1]alkanones via a cationic reaction cascade. AB - [reaction: see text] The acid-mediated Prins/pinacol and the triple domino reactions Diels-Alder/Prins/pinacol were used to construct highly functionalized bicyclo[m.n.1]alkanones 19-29 and 33a-c possessing various ring sizes from ketals 8-18 and 31a-c in 44-96% yields. This approach proves to be highly efficient and reliable to generate high molecular complexity in a single step. PMID- 16354102 TI - Diagnosis and management of work-related asthma. PMID- 16354103 TI - The new International Health Regulations and the federalism dilemma. PMID- 16354104 TI - Perspectives on the efficacy of antidepressants for child and adolescent depression. PMID- 16354105 TI - Endothelial lipase concentrations are increased in metabolic syndrome and associated with coronary atherosclerosis. AB - BACKGROUND: Endothelial lipase (EL), a new member of the lipase family, has been shown to modulate high-density lipoprotein (HDL-C) metabolism and atherosclerosis in mouse models. We hypothesized that EL concentrations would be associated with decreased HDL-C and increased atherosclerosis in humans. METHODS AND FINDINGS: Healthy individuals with a family history of premature coronary heart disease (n = 858) were recruited as part of the Study of the Inherited Risk of Atherosclerosis. Blood was drawn in the fasting state before and, in a subgroup (n = 510), after administration of a single dose of intravenous heparin. Plasma lipids were measured enzymatically, lipoprotein subclasses were assessed by nuclear magnetic resonance, and coronary artery calcification (CAC) was quantified by electron beam computed tomography. Plasma EL mass was measured using a newly developed enzyme-linked immunosorbent assay. Median EL mass in pre heparin plasma was 442 (interquartile range = 324-617) ng/ml. Median post-heparin mass was approximately 3-fold higher, 1,313 (888-1,927) ng/ml. The correlation between pre-heparin EL mass and post-heparin EL mass was 0.46 (p < 0.001). EL mass concentrations in both pre- and post-heparin plasma significantly correlated with all NCEP ATPIII-defined metabolic syndrome factors: waist circumference (r = 0.28 and 0.22, respectively, p < 0.001 for each), blood pressure (r = 0.18 and 0.24, p < 0.001 for each), triglycerides (r = 0.22, p < 0.001; and 0.13, p = 0.004), HDL cholesterol (r = -0.11, p = 0.002; and -0.18, p < 0.001), and fasting glucose (r = 0.11 and 0.16, p = 0.001 for both). EL mass in both routine (odds ratio [OR] = 1.67, p = 0.01) and post-heparin (OR = 2.42, p = 0.003) plasma was associated with CAC as determined by ordinal regression after adjustment for age, gender, waist circumference, vasoactive medications, hormone replacement therapy (women), and established cardiovascular risk factors. CONCLUSIONS: We report, to our knowledge for the first time, that human plasma EL concentrations, in both post-heparin and routine pre-heparin plasma, are significantly associated with metabolic syndrome features and with subclinical atherosclerosis. EL may be a pro atherogenic factor in humans, especially in overweight individuals and those with metabolic syndrome. PMID- 16354106 TI - Analysis of prion strains by PrPSc profiling in sporadic Creutzfeldt-Jakob disease. AB - BACKGROUND: Prion diseases are a group of invariably fatal neurodegenerative disorders affecting humans and a wide range of mammals. An essential part of the infectious agent, termed the prion, is composed of an abnormal isoform (PrPSc) of a host-encoded normal cellular protein (PrPC). The conversion of PrPC to PrPSc is thought to play a crucial role in the development of prion diseases and leads to PrPSc deposition, mainly in the central nervous system. Sporadic Creutzfeldt Jakob disease (sCJD), the most common form of human prion disease, presents with a marked clinical heterogeneity. This diversity is accompanied by a molecular signature which can be defined by histological, biochemical, and genetic means. The molecular classification of sCJD is an important tool to aid in the understanding of underlying disease mechanisms and the development of therapy protocols. Comparability of classifications is hampered by disparity of applied methods and inter-observer variability. METHODS AND FINDINGS: To overcome these difficulties, we developed a new quantification protocol for PrPSc by using internal standards on each Western blot, which allows for generation and direct comparison of individual PrPSc profiles. By studying PrPSc profiles and PrPSc type expression within nine defined central nervous system areas of 50 patients with sCJD, we were able to show distinct PrPSc distribution patterns in diverse subtypes of sCJD. Furthermore, we were able to demonstrate the co-existence of more than one PrPSc type in individuals with sCJD in about 20% of all patients and in more than 50% of patients heterozygous for a polymorphism on codon 129 of the gene encoding the prion protein (PRNP). CONCLUSION: PrPSc profiling represents a valuable tool for the molecular classification of human prion diseases and has important implications for their diagnosis by brain biopsy. Our results show that the co-existence of more than one PrPSc type might be influenced by genetic and brain region-specific determinants. These findings provide valuable insights into the generation of distinct PrPSc types. PMID- 16354109 TI - UV radiation, vitamin D and human health: an unfolding controversy introduction. PMID- 16354108 TI - Prospective study of one million deaths in India: rationale, design, and validation results. AB - BACKGROUND: Over 75% of the annual estimated 9.5 million deaths in India occur in the home, and the large majority of these do not have a certified cause. India and other developing countries urgently need reliable quantification of the causes of death. They also need better epidemiological evidence about the relevance of physical (such as blood pressure and obesity), behavioral (such as smoking, alcohol, HIV-1 risk taking, and immunization history), and biological (such as blood lipids and gene polymorphisms) measurements to the development of disease in individuals or disease rates in populations. We report here on the rationale, design, and implementation of the world's largest prospective study of the causes and correlates of mortality. METHODS AND FINDINGS: We will monitor nearly 14 million people in 2.4 million nationally representative Indian households (6.3 million people in 1.1 million households in the 1998-2003 sample frame and 7.6 million people in 1.3 million households in the 2004-2014 sample frame) for vital status and, if dead, the causes of death through a well validated verbal autopsy (VA) instrument. About 300,000 deaths from 1998-2003 and some 700,000 deaths from 2004-2014 are expected; of these about 850,000 will be coded by two physicians to provide causes of death by gender, age, socioeconomic status, and geographical region. Pilot studies will evaluate the addition of physical and biological measurements, specifically dried blood spots. Preliminary results from over 35,000 deaths suggest that VA can ascertain the leading causes of death, reduce the misclassification of causes, and derive the probable underlying cause of death when it has not been reported. VA yields broad classification of the underlying causes in about 90% of deaths before age 70. In old age, however, the proportion of classifiable deaths is lower. By tracking underlying demographic denominators, the study permits quantification of absolute mortality rates. Household case-control, proportional mortality, and nested case control methods permit quantification of risk factors. CONCLUSIONS: This study will reliably document not only the underlying cause of child and adult deaths but also key risk factors (behavioral, physical, environmental, and eventually, genetic). It offers a globally replicable model for reliably estimating cause specific mortality using VA and strengthens India's flagship mortality monitoring system. Despite the misclassification that is still expected, the new cause-of death data will be substantially better than that available previously. PMID- 16354107 TI - Interleukin-1 stimulates beta-cell necrosis and release of the immunological adjuvant HMGB1. AB - BACKGROUND: There are at least two phases of beta-cell death during the development of autoimmune diabetes: an initiation event that results in the release of beta-cell-specific antigens, and a second, antigen-driven event in which beta-cell death is mediated by the actions of T lymphocytes. In this report, the mechanisms by which the macrophage-derived cytokine interleukin (IL) 1 induces beta-cell death are examined. IL-1, known to inhibit glucose-induced insulin secretion by stimulating inducible nitric oxide synthase expression and increased production of nitric oxide by beta-cells, also induces beta-cell death. METHODS AND FINDINGS: To ascertain the mechanisms of cell death, the effects of IL-1 and known activators of apoptosis on beta-cell viability were examined. While IL-1 stimulates beta-cell DNA damage, this cytokine fails to activate caspase-3 or to induce phosphatidylserine (PS) externalization; however, apoptosis inducers activate caspase-3 and the externalization of PS on beta cells. In contrast, IL-1 stimulates the release of the immunological adjuvant high mobility group box 1 protein (HMGB1; a biochemical maker of necrosis) in a nitric oxide-dependent manner, while apoptosis inducers fail to stimulate HMGB1 release. The release of HMGB1 by beta-cells treated with IL-1 is not sensitive to caspase-3 inhibition, while inhibition of this caspase attenuates beta-cell death in response to known inducers of apoptosis. CONCLUSIONS: These findings indicate that IL-1 induces beta-cell necrosis and support the hypothesis that macrophage derived cytokines may participate in the initial stages of diabetes development by inducing beta-cell death by a mechanism that promotes antigen release (necrosis) and islet inflammation (HMGB1 release). PMID- 16354110 TI - Daily duration of vitamin D synthesis in human skin with relation to latitude, total ozone, altitude, ground cover, aerosols and cloud thickness. AB - Vitamin D production in human skin occurs only when incident UV radiation exceeds a certain threshold. From simulations of UV irradiances worldwide and throughout the year, we have studied the dependency of the extent and duration of cutaneous vitamin D production in terms of latitude, time, total ozone, clouds, aerosols, surface reflectivity and altitude. For clear atmospheric conditions, no cutaneous vitamin D production occurs at 51 degrees latitude and higher during some periods of the year. At 70 degrees latitude, vitamin D synthesis can be absent for 5 months. Clouds, aerosols and thick ozone events reduce the duration of vitamin D synthesis considerably, and can suppress vitamin D synthesis completely even at the equator. A web page allowing the computation of the duration of cutaneous vitamin D production worldwide throughout the year, for various atmospheric and surface conditions, is available on the Internet at http://zardoz.nilu.no/~olaeng/fastrt/VitD.html and http://zardoz.nilu.no/~olaeng/fastrt/VitD-ez.html. The computational methodology is outlined here. PMID- 16354111 TI - Youth solar ultraviolet radiation exposure, concurrent activities and sun protective practices: a review. AB - To assist standardization of procedures, facilitate comparisons, and help guide research efforts to optimally inform development of appropriately targeted interventions, there is a need to review methods used to quantify child and adolescent solar ultraviolet radiation (UV) exposure, related outdoor activities and sun-protective practices. This holistic approach is essential for comprehensive research that will provide all-inclusive, informative and meaningful messages for preventive measures of harmful UV exposure. Two databases were searched and 29 studies were retrieved, and these studies report measurement or assessment techniques documenting UV exposure patterns and related outdoor activities. Polysulfone film badges were the main measurement instrument used in 10 studies, with questionnaire, survey data, observation, a model, electronic dosimeters, biological dosimeters, colorimeter and UV colouring labels used in the remaining studies. Methods used to record activities included self-report, parental report, a logbook and observation. Measurement duration and unit of UV exposure varied in most studies, but a method common to 15 studies was measured UV exposure as a percentage of ambient UV. The studies reviewed do not provide sufficient information for the development and evaluation of targeted youth sun protection programs. Studies are required which document precise UV exposure, concurrent activities and sun protection usage for children and adolescents. PMID- 16354112 TI - Rapid detection of Aspergillus flavus contamination in peanut with novel delayed luminescence spectra. AB - Employing a luminescence spectrometer the usefulness of light-induced delayed luminescence (DL) for the detection of aflatoxinB1 (AfB1) contamination in whole peanut was studied. Peanut was artificially contaminated with Aspergillus flavus (7.5 x 10(5)conidia/mL) and incubated for 0-72 h. The DL and fluorescence spectra of contaminated peanut were obtained by a luminescence spectrometer. The correlation between the spectra and contamination levels was established. The DL and fluorescence intensity has a negative correlation with the AfB1 concentration. Our results suggested that the DL technique might be useful for the rapid and noninvasive evaluation of AfB1 contamination levels in whole peanut. PMID- 16354113 TI - Vibrational spectroscopic studies on the disulfide formation and secondary conformational changes of captopril-HSA mixture after UV-B irradiation. AB - The effects of pH and ultraviolet-B (UV-B) irradiation on the secondary structure of human serum albumin (HSA) in the absence or presence of captopril were investigated by an attenuated total reflection (ATR)/Fourier transform infrared (FTIR) spectroscopy. The UV-B exposure affecting the stability of captopril before and after captopril-HSA interaction was also examined by using confocal Raman microspectroscopy. The results indicate that the transparent pale-yellow solution for captopril-HSA mixture in all pH buffer solutions, except pH 5.0 approximately 7.0, changed into a viscous form then a gel form with UV-B exposure time. The secondary structural transformation of HSA in the captopril-HSA mixture with or without UV-B irradiation was found to shift the maxima amide I peak in IR spectra from 1652 cm(-1) assigned to alpha-helix structure to 1622 cm(-1) because of a beta-sheet structure, which was more evident in pH 3.0, 8.0 or 9.0 buffer solutions. The Raman shift from 1653 cm(-1) (alpha-helix) to 1670 cm(-1) (beta sheet) also confirmed this result. Captopril dissolved in distilled water with or without UV-B irradiation was determined to form a captopril disulfide observed from the Raman spectra of 512 cm(-1), which was exacerbated by UV-B irradiation. There was little disulfide formation in the captopril-HSA mixture even with long term UV-B exposure, but captopril might interact with HSA to change the protein secondary structure of HSA whether there was UV-B irradiation or not. The pH of the buffer solution and captopril-HSA interaction may play more important roles in transforming the secondary structure of HSA from alpha-helix to beta-sheet in the corresponding captopril-HSA mixture than UV-B exposure. The present study also implies that HSA has the capability to protect the instability of captopril in the course of UV-B irradiation. In addition, a partial unfolding of HSA induced by pH or captopril-HSA interaction under UV-B exposure is proposed. PMID- 16354114 TI - Hypericin-mediated photocytotoxic effect on HT-29 adenocarcinoma cells is reduced by light fractionation with longer dark pause between two unequal light doses. AB - The present study demonstrates the in vitro effect of hypericin-mediated PDT with fractionated light delivery. Cells were photosensitized with unequal light fractions separated by dark intervals (1 or 6 h). We compared the changes in viability, cell number, survival, apoptosis and cell cycle on HT-29 cells irradiated with a single light dose (12 J/cm(2)) to the fractionated light delivery (1 + 11 J/cm(2)) 24 and 48 h after photodynamic treatment. We found that a fractionated light regime with a longer dark period resulted in a decrease of hypericin cytotoxicity. Both cell number and survival were higher after light sensitization with a 6-h dark interval. DNA fragmentation occurred after a single light-dose application, but in contrast no apoptotic DNA formation was detected with a 6-h dark pause. After fractionation the percentage of cells in the G1 phase of the cell cycle was increased, while the proportion of cells in the G2 phase decreased as compared to a single light-dose application, i.e. both percentage of cells in the G1 and G2 phase of the cell cycle were near control levels. We presume that the longer dark interval after the irradiation of cells by first light dose makes them resistant to the effect of the second illumination. These findings confirm that the light application scheme together with other photodynamic protocol components is crucial for the photocytotoxicity of hypericin. PMID- 16354116 TI - Phototaxis in the cyanobacterium Synechocystis sp. PCC 6803: role of different photoreceptors. AB - The second cyanobacterial phytochrome Cph2 from Synechocystis sp. PCC 6803 was suggested as a part of a light-stimulated signal transduction chain inhibiting movement toward blue light. Cph2 has the two bilin binding sites, cysteine-129 and cysteine-1022, that might be involved in sensing of red/far-red and blue light, respectively. Here, we present data on wavelength dependence of the phototaxis inhibition under blue light, indicating that Cph2 itself is the photoreceptor for this blue light response. We found that inhibition of blue light phototaxis in wild-type cells occurred below the transition point of about 470 nm. Substitution of cysteine-1022 with valine led to photomovement of the cells toward blue light (cph2(-) mutant phenotype). Analysis of mutants lacking cysteine-129 in the N-terminal chromophore binding domain indicated that this domain is also important for Cph2 function or folding of the protein. Furthermore, putative blue-light and phytochrome-like photoreceptors encoded by the Synechocystis sp. PCC 6803 genome were inactivated in wild-type and cph2 knockout mutant background. Our results suggest that none of these potential photoreceptors interfere with Cph2 function, although inactivation of taxD1 as well as slr1694 encoding a BLUF protein led to cells that reversed the direction of movement under blue light illumination in mutant strains of cph2. PMID- 16354115 TI - The detection of UV-induced membrane damages by a combination of two biosensor techniques. AB - Ketoprofen is an important anti-inflammatory drug, but its dermal application is limited because of the photosensitizing properties causing phototoxic reactions of the skin when exposed to UV light. We have recently investigated the peroxide formation of ketoprofen in solutions of linoleic acid during UV irradiation. To continue these studies and focus on UV-induced changes in membrane integrity and barrier function we established an in vitro model using two biosensor techniques simultaneously. Support-fixed bilayers were irradiated with different doses of UV B up to damaging intensities with or without ketoprofen. Cyclic voltammetry was carried out to detect alterations in membrane permeability; quartz crystal microbalance (QCM) measurements were helpful in analyzing whether a permeability increase was caused by depletion of membrane components. In absence of ketoprofen, increasing UV-B doses induce membrane permeabilities of both unsaturated and saturated bilayers. QCM measurements could not reveal a significant loss of membrane components as a reason for the permeability. In contrast, 0.3 mM ketoprofen induced a dose-dependent increase in membrane permeability. QCM results indicated a mass loss. Although this model does not explain all molecular mechanisms of membrane damage by ketoprofen, the combined application of both QCM and CV is a novel and powerful tool to investigate functional mechanisms of UV-induced membrane damages. PMID- 16354117 TI - Protection against photoaging in the hairless mouse by the isoflavone equol. AB - Topical application of the isoflavone equol immediately following solar-simulated UV (SSUV) radiation exposure has previously been demonstrated to have significant photoprotective effects. Equol reduced both the inflammatory edema and the systemic suppression of the contact hypersensitivity reaction in hairless mice. Furthermore, daily topical equol application immediately following irradiation during a 10-week chronic SSUV exposure regime also reduced photocarcinogenesis severity in the mouse. This study examines the potential for topical equol to prevent photoaging in response to chronic SSUV irradiation for up to 30 weeks. We did not find consistent expression of the characteristic markers of photoaging until 30 weeks, although moderate epidermal hyperplasia and a transient increase in dermal mast cell numbers were evident after 1 week. Daily application of 10 muM equol lotion significantly reduced these early changes. However after 30 weeks of SSUV exposure, photoaging was well developed, as shown histologically by markedly increased epidermal hyperplasia, increased dermal mast cell number, pronounced focal elastotic deposits, degraded dermal collagen and deposition of glycosaminoglycans in the lower dermis. Topical equol treatment protected significantly from each of these impairments, as demonstrated histologically and quantitatively. Additionally, equol was found to have strong antioxidant action against acute UVA (320-400 nm)-induced lipid peroxidation of mouse skin, this property accounting for its antiphotoaging mechanism. The evidence for equol's antiphotoaging activity, taken together with its anti-inflammatory, immunoprotective and anticarcinogenic efficacy against SSUV irradiation in the mouse, suggests that equol could be developed as a helpful topical photoprotective agent for daily use by humans. PMID- 16354118 TI - Is primary care ready for pharmacogenetics? PMID- 16354119 TI - Prospective care: a personalized, preventative approach to medicine. PMID- 16354121 TI - Predicting and diagnosing abacavir and nevirapine drug hypersensitivity: from bedside to bench and back again. AB - There is a growing discussion surrounding the issue of personalized approaches to drug prescription based on an individual's genetic makeup. This field of investigation has focused primarily on identifying genetic factors that influence drug metabolism and cellular disposition, thereby contributing to dose-dependent toxicities and/or variable drug efficacy. However, pharmacogenetic approaches have also proved valuable in predicting drug hypersensitivity reactions in selected patient populations, including HIV-infected patients receiving long-term antiretroviral therapy. In this instance, susceptibility has been strongly linked to genetic loci involved in antigen recognition and presentation to the immune system--most notably within the major histocompatibility complex (MHC) region- consistent with the notion that hypersensitivity reactions represent drug specific immune responses that are largely dose independent. Here the authors describe their experiences with the development of pharmacogenetic approaches to hypersensitivity reactions associated with abacavir and nevirapine, two commonly prescribed antiretroviral drugs. It is demonstrated that prospective screening tests to identify and exclude individuals with a certain genetic makeup may be largely successful in decreasing or eliminating incidence of these adverse drug reactions in certain populations. This review also explores the broader implications of these findings. PMID- 16354122 TI - Copy number variants and pharmacogenomics. AB - The earliest pharmacogenomic studies focused on highly penetrant sequence polymorphisms in drug-metabolizing enzymes. The recent discovery of the widespread occurrence of copy number variants/polymorphisms in the human genome holds promise for new pharmacogenomic discoveries, aside from the commonly used single nucleotide polymorphism approach. Here we review the discovery of copy number variants and speculate on their implications for pathophysiology and pharmacogenomics. PMID- 16354123 TI - Pharmacogenomics: a path to predictive medicine for schizophrenia. AB - A significant variability is observed among patients in response to antipsychotics, and is caused by a variety of factors. This review summarizes the available knowledge of associations between pharmacogenetics and drug response in schizophrenia. The multifactorial etiology of schizophrenia makes it a complex interaction of symptoms. Adopting a pharmacogenomics approach represents a unique opportunity for the prediction of response to antipsychotic drugs by investigating genes implicated with specific symptoms and side effects. A network model of the interaction/crosstalk between the neurotransmitter signaling systems is presented to emphasize the importance of the genes associated with the molecular mechanisms of the disease and drug response. These genes may serve as potential susceptibility genes and drug targets for schizophrenia. The crucial point for the identification of a significant biologic marker(s) will include not only the experimental validation of the genes involved in the neurotransmitter signaling systems, but also the availability of large exactly comparable phenotyped patients samples. Coupling our knowledge of genetic polymorphisms with clinical response data promises a bright future for rapid advances in personalized medicine. PMID- 16354124 TI - Patients' and physicians' perspectives on pharmacogenetic testing. AB - INTRODUCTION: The integration of pharmacogenetic testing into routine care will, in part, depend upon the patients' and physicians' acceptance of these tests. Empirical data regarding patients' and physicians' views on pharmacogenetic testing are lacking. OBJECTIVES: To explore patients' and physicians' perspectives on the potential implications of pharmacogenetic testing, particularly focusing on asthma, and to analyze the possible determinants of their expectations, hopes and fears. METHODS: We conducted telephone interviews with patients with asthma or chronic obstructive pulmonary disease taking part in a larger pharmacogenetic study, in addition to general practitioners (GPs) from a different region in Germany. A total of 328 patients and 378 GPs were invited to participate. Determinants of their attitudes toward pharmacogenetic testing were assessed using logistic regression analysis. RESULTS: Informed consent to participate in this study was given by 196 patients (60%) and 106 GPs (28%). Most patients (96%) and physicians (52%) appreciated the availability of pharmacogenetic tests for a disease such as asthma. Approximately a third of the patients worried about potential unfavorable test results (35%) and violation of privacy (36%). Female patients were more likely to have a fearful attitude (odds ratio [OR]=2.85; 95% confidence interval [CI]=1.58-5.12). Younger patients were generally more likely to be hopeful about the usefulness of pharmacogenetic testing (OR=2.12; CI=1.01-4.46). The GPs' concerns were mainly related to the possibility that patients might either be put under pressure to be tested (72%) or be disadvantaged at private health insurance agencies (61%). The nature of the responsible institution, the clarity of the research aim and explicit informed consent from patients influenced a physicians' decision regarding whether to support a pharmacogenetic study. CONCLUSION: The concerns of patients and GPs differ somewhat with respect to negative psychosocial consequences, discrimination or violation of privacy. Development of information for physicians and patients would be helpful in preventing unrealistic fears or hopes. PMID- 16354125 TI - Defining the opportunity for pharmacogenetic intervention in primary care. AB - Pharmacogenetics (PG), the study of human genome function and its effects on drug response, represents an exciting approach for reducing adverse drug events and increasing therapeutic efficacy. However, there is no clear information of the potential impact of PG in the primary care setting. Therefore, a study was conducted to determine the frequency of use of medications under PG influence, including 16 PG adverse drug reaction (ADR)-associated medications, in the primary care setting. PATIENTS AND METHODS: A cohort of 607 consecutive patients was accrued over a 3-month period from three primary care practices. Patients were asked to answer a verbal survey of demographics and medication use during the past 12 months. The survey specifically evaluated 16 drugs known to commonly cause ADRs and undergo metabolism by polymorphic enzymes. Patients also disclosed information on all other medication use in the last year. Medication use was verified by chart review. The primary outcome was the frequency of medication use. RESULTS: Among the 16 ADR-associated medications, patients used analgesics (88.5%), antihypertensives (14.3%) and antidepressants (9.6%) most commonly. Overall, 28.6% of patients took more than one of the PG ADR-associated medications. Neither gender nor race appeared to influence the frequency of use of these medications (p=0.5 and p=0.08, respectively). Patients taking one or more of the drugs were older (p<0.001). More patients seen for a chronic visit took one or more of the ADR-associated drugs than patients seen for an acute visit (35.8 versus 18.5%, p<0.001). DISCUSSION: This is the first attempt to describe the potential role of pharmacogenetics in the primary care setting. The findings indicate that at least one in four primary care patients take at least one medication that commonly causes adverse drug reactions due to genetic variability in drug metabolism, indicating that there is a potential role of pharmacogenomics in primary care. Nearly every patient was on a medication with putative PG association. CONCLUSIONS: Studies of the ability of PG should not be limited to medical subspecialties, as there is a great potential impact of PG on the primary care setting. PMID- 16354126 TI - Genomic profiling associated with recurrence in patients with rectal cancer treated with chemoradiation. AB - PURPOSE: Stage II and III adenocarcinoma of the rectum has an overall 5-year survival rate of approximately 50%, and tumor recurrence remains a major problem despite an improvement in local control through chemotherapy and radiation. The efficacy of chemoradiation therapy may be significantly compromised as a result of interindividual variations in clinical response and host toxicity. Therefore, it is imperative to identify those patients who will benefit from chemoradiation therapy and those who will develop recurrent disease. In this study, we tested whether a specific pattern of 21 polymorphisms in 18 genes involved in the critical pathways of cancer progression (i.e., drug metabolism, tumor microenvironment, cell cycle regulation, and DNA repair) will predict the risk of tumor recurrence in rectal cancer patients treated with chemoradiation. PATIENTS AND METHODS: A total of 90 patients with Stage II or III rectal cancer treated with chemoradiation were genotyped using polymerase chain reaction (PCR)-based techniques for 21 polymorphisms. RESULTS: A polymorphism in interleukin (IL)-8 was individually associated with risk of recurrence. Classification and regression tree analysis of all polymorphisms and clinical variables developed a risk tree including the following variables: node status, IL-8, intracellular adhesion molecule-1, transforming growth factor-beta, and fibroblast growth factor receptor 4. CONCLUSION: Genomic profiling may help to identify patients who are at high risk for developing tumor recurrence, and those who are more likely to benefit from chemoradiation therapy. A larger prospective study is needed to validate these preliminary data using germline polymorphisms on tumor recurrences in rectal cancer patients treated with chemoradiation. PMID- 16354127 TI - Challenges and opportunities in the application of pharmacogenetics to antiepileptic drug therapy. AB - The recent surge of interest in pharmacogenetics has provoked considerable thought regarding its relevance to antiepileptic drug (AED) therapy. Initial studies have focused on genes whose products play a putatively important role in AED pharmacology, particularly drug transporter proteins, drug metabolizing enzymes and ion channel subunits. However, there is a lack of good correspondence between results from different laboratories, and more recent findings are awaiting attempts at confirmation. Thus, there are currently no AED treatment guidelines that are informed by pharmacogenetic data. In order to begin to have clinical impact, standards specific to the conduct of future AED studies must be established. Of particular importance are the need for accurate epilepsy classification, appropriate AED selection and clear and objective assessment outcome measures. In addition, general standards for analysis and interpretation of genetic association data must be better codified and applied consistently across studies. Finally, extensive clinical research networks must be formulated and large numbers of well characterized patients must be recruited. Further development of these critical factors will optimize chances for overcoming current challenges posed by AED pharmacogenetic research and ultimately allow the realization of improved, more rational therapeutic strategies. PMID- 16354128 TI - Expression biomarkers for clinical efficacy and outcome prediction in cancer. AB - Progress in cancer treatment has been slow, and the outlook for curing cancer is only marginally different from the situation a decade ago. Paradoxically, although the pharmaceutical industry has stepped up costly discovery research and drug development, approvals are on the decline and pipelines are dwindling. In an effort to reduce the number of drug failures and curtail burgeoning R&D costs, drug companies are exploring the use of biomarkers to evaluate toxicity and efficacy earlier in the development process. Biomarkers hold promise for optimization in dosing, adverse event prediction, efficacy evaluation, lead prioritization, and mechanism-of-action profiling of drug candidates. Furthermore, clinicians can use biomarkers to monitor patient response in clinical trials. In this perspective article, the authors explore the applications of cancer-related expression biomarkers in drug discovery and discuss how this will impact the industry and benefit the patient. PMID- 16354129 TI - Bridging the divide: the need for translational informatics. AB - Translational science promises to deliver real benefit to the pharmaceutical industry, reducing attrition and affording high quality, efficacious medicines. The development and use of biomarkers aims to reduce drug development risks and generate a better understanding of disease. Informatics is an essential component of the translational science toolkit; researchers must be able to work effectively with biomarker data, and the capture and reuse of knowledge is vital for long-term success. An analysis of current data and knowledge management practices in the translational science area is presented. PMID- 16354130 TI - Microarray expression technology: from start to finish. AB - The recent introduction of new microarray expression technologies and the further development of established platforms ensure that the researcher is presented with a range of options for performing an experiment. Whilst this has opened up the possibilities for future applications, such as exon-specific arrays, increased sample throughput and 'chromatin immunoprecipitation (ChIP) on chip' experiments, the initial decision processes and experiment planning are made more difficult. This review will give an overview of the various technologies that are available to perform a microarray expression experiment, from the initial planning stages through to the final data analysis. Both practical aspects and data analysis options will be considered. The relative advantages and disadvantages will be discussed with insights provided for future directions of the technology. PMID- 16354132 TI - Examining the influence of using same versus different questions on the reliability of the medical school preadmission interview. AB - BACKGROUND: Researchers generally recommend a structured format for the medical school preadmission interview (MSPI). However, the relative benefits of various elements of structure remain unexamined. PURPOSE: In this study, we compared the performance of a highly structured interview format with a semistructured format. Specifically, we examined how the reliability of interview ratings is likely to change when using the same versus different questions for each applicant being interviewed. METHOD: Variance components from a generalizability (G) study of a structured interview are used in decision studies to compare the relative efficiency of using the same versus different questions for each applicant. RESULTS: Using different questions for each interviewee is practically as reliable as using the same questions for all applicants (G = .55 vs. .57, respectively). CONCLUSIONS: Because there are a number of drawbacks to using the same questions for all applicants (i.e., security and validity) and little advantage in terms of increased reliability, the semistructured question format should be considered when conducting the MSPI. A suggested method of implementing a semistructured interview is by presenting each applicant a set of questions randomly drawn from a pool of interview questions. PMID- 16354133 TI - The relationship between service learning and medical student academic and professional outcomes. AB - BACKGROUND: Little is known about the relationship between medical student participation in service and performance outcomes. PURPOSE: To examine relationships between involvement in required service learning and measures of academic and professional achievement and specialty choice. METHODS: Using preexisting, self-reported data for Tulane University School of Medicine's class of 2004, we categorized students as low, moderate, or high on commitment and involvement in service learning (CISL) and compared to multiple outcomes: class rank; election to the Alpha Omega Alpha (AOA) Honor Society; election to the Gold Humanism in Medicine Honor Society; total percent scores on 2 multistation, structured, clinical examinations; and specialty choice per 2004 residency match results. RESULTS: Service learning involvement was related to class rank. There was a marginal relationship between CISL and election to AOA but no association with election to the Gold Humanism in Medicine Honor Society, Year 2 or 3 clinical skills examination results, or specialty choice. CONCLUSIONS: High participation in service learning and service learning leadership identify a group of students less likely to be in the top quartile of their class yet who are making important contributions to the community and profession. PMID- 16354134 TI - Teaching cultural competency through narrative medicine: intersections of classroom and community. AB - BACKGROUND: Cultural competency and narrative medicine are perspectives that assist medical educators in teaching effective, empathetic communication and service delivery to a variety of patients. PURPOSE/METHODS: In this article, we describe a unique educational activity at the crossroads of these perspectives in which pediatric residents participated in a monthly reading and discussion group with staff members of an inner-city Dominican American community organization. RESULTS: By discussing a literary text rather than cases and facilitating discussions with particular attention to power, not only were historic conflicts between the groups circumvented, but an environment was created in which discussants drew heavily from personal and professional experiences. Qualitative evaluation of both groups revealed improved self-reported understanding of (a) issues of cultural diversity, (b) issues of medical culture, and (c) physicians' attitudes and behaviors in practice. CONCLUSION: Methodologies drawing from cultural competency and narrative medicine can be used to help physicians work in multidisciplinary, multicultural teams in and out of the medical institution. PMID- 16354135 TI - Development and evaluation of an interactive multimedia clinical skills teaching program designed for the pediatric clerkship. AB - BACKGROUND AND PURPOSE: We evaluated the physical-examination section of a multimedia program developed to teach infant history and physical-examination skills. METHODS: A total of 71 students participated: one group viewed only the physical-examination section (PX), one the history section (HX), one none of the program (CX). We assessed physical-examination skills by direct observation of medical students performing an abdominal exam and scored using a checklist at baseline, immediately after intervention, and at the end of the pediatric clerkship. We analyzed results using analysis of variance with repeated measures. RESULTS: Baseline scores were PX = 2.5, HX = 2.8. The PX group scored significantly higher immediately postintervention at 6.8 compared to the HX group (3.1). At the end of the clerkship, significant differences between the groups remained. Final group mean scores were PX = 5.5, HX = 4.4, and CX = 2.7. CONCLUSION: The program improved examination skills with attenuation over 6 weeks. PMID- 16354136 TI - The cognitive validity of the Script Concordance Test: a processing time study. AB - BACKGROUND: According to the theory on which the Script Concordance Test (SCT) is based, scripts contain expectations on features that are associated with each illness and about the range of values that are typical, atypical, or incompatible. PURPOSE: To document the construct validity of the SCT, we investigated the theory prediction that once a script is activated, new incoming information (e.g., additional clinical features) is processed faster if it is typical for that script than if it is atypical. If it is incompatible, processing time falls in between. METHODS: We presented 2 groups of participants (30 fourth year medical students and 30 full-time geriatricians) with 64 clinical vignettes (divided over 5 types of prevalent clinical presentations in geriatrics), each accompanied by a diagnostic hypothesis aimed to instantiate an appropriate script. Next, we presented a new finding, which could be typical, atypical, or incompatible given the hypothesis. Participants had to decide as quickly and accurately as possible whether the new finding increased, decreased, of did not affect the likelihood of the diagnostic hypothesis. We administered the test on a computer. The dependent variable was processing time. We analyzed data with a repeated measure 2 x 3 analysis of variance. RESULTS: Typical information was processed faster than atypical and incompatible information (M = 10.6 sec vs. 19.2 sec and 16.4 sec, respectively; p lt; .001 for both). Incompatible information was processed faster than atypical information (16.4 sec vs. 19.2; p < .001). There was no significant difference between the groups of geriatricians and students. CONCLUSION: It is possible to predict what kind of information will be processed faster depending of the typicality and compatibility of clinical data for given hypotheses. Results support SCT construct validity. PMID- 16354137 TI - Medical education and the ACGME duty hour requirements: assessing the effect of a day float system on educational activities. AB - BACKGROUND: In July 2003, the Accreditation Council for Graduate Medical Education (ACGME) instituted new resident work hour mandates, which are being shown to improve resident well-being and patient safety. However, there are limited data on the impact these new mandates may have on educational activities. PURPOSES: To assess the impact on educational activities of a day float system created to meet ACGME work hour mandates. METHODS: The inpatient ward coverage was changed by adding a day float team responsible for new patient admissions in the morning, with the on-call teams starting later and being responsible for new patient admissions thereafter. I surveyed the residents to assess the impact of this new system on educational activities-resident autonomy, attending teaching, conference attendance, resident teaching, self-directed learning, and ability to complete patient care responsibilities. RESULTS: There was no adverse effect of the day float system on educational activities. House staff reported increased autonomy, enhanced teaching from attending physicians, and improved ability to complete patient care responsibilities. Additionally, house staff demonstrated improved compliance with the ACGME mandates. CONCLUSIONS: The implementation of a novel day float system for the inpatient medicine ward service improved compliance with ACGME work duty requirements and did not adversely impact educational activities of the residency training program. PMID- 16354138 TI - Enhancing the effectiveness of One-Minute Preceptor faculty development workshops. AB - BACKGROUND: The One Minute Preceptor (OMP) faculty development workshop is aimed at improving outpatient preceptors' abilities to teach in ambulatory settings. Practice applying the 5 microskills is central to learning the OMP, yet participants frequently avoid the role-playing portion of the standard workshop. Task complexity may result in significant cognitive burden while learning these new teaching skills. DESCRIPTION: The development of highly scripted case scenarios and standardized learners (SLs) to role-play authentic teaching situations are aimed at reducing task complexity. We describe case development, SL training, and application of these enhancements in 5 OMP workshops. EVALUATION: Qualitative assessments and workshop participant evaluations showed improved role-playing participation and satisfaction with the authenticity of the training sessions. CONCLUSION: The introduction of highly scripted cases and SLs appear to effectively help participants overcome dissatisfaction with role playing and provide increasingly challenging yet authentic clinical teaching scenarios for skills practice. We recommend further research using objective measures. PMID- 16354140 TI - Is medicine altruistic? A query from the medical school admissions office. AB - BACKGROUND: Applicants to medical school often state that they are motivated by a desire to help others. Admissions officers must evaluate these claims, but assessment of altruism is difficult and imprecise. SUMMARY: A purely utilitarian moral philosophy denies the possibility of altruism. However, the Enlightenment philosopher Hume described a force, sympathy, that engages channels of communication between persons and rewards benevolence. Although there are many pretenders to altruism, the medical school interview provides an opportunity to create a channel of sympathy and to assess its authenticity. CONCLUSIONS: By probing for evidence of clinical sympathy, the medical school interviewer may be able to verify whether the applicant's claims of altruism are authentic. Claims of altruism that have not been borne out in action should be viewed with skepticism. PMID- 16354139 TI - A model for educational feedback based on clinical communication skills strategies: beyond the "feedback sandwich". AB - BACKGROUND: Feedback is an essential tool in medical education, and the process is often difficult for both faculty and learner. There are strong analogies between the provision of educational feedback and doctor-patient communication during the clinical encounter. DESCRIPTION: Relationship-building skills used in the clinical setting-Partnership, Empathy, Apology, Respect, Legitimation, Support (PEARLS)-can establish trust with the learner to better manage difficult feedback situations involving personal issues, unprofessional behavior, or a defensive learner. Using the stage of readiness to change (transtheoretical) model, the educator can "diagnose" the learner's stage of readiness and employ focused interventions to encourage desired changes. EVALUATION: This approach has been positively received by medical educators in faculty development workshops. CONCLUSIONS: A model for provision of educational feedback based on communication skills used in the clinical encounter can be useful in the medical education setting. More robust evaluation of the construct validity is required in actual training program situations. PMID- 16354141 TI - Procedures for establishing defensible absolute passing scores on performance examinations in health professions education. AB - BACKGROUND: Establishing credible, defensible, and acceptable passing scores for performance-type examinations in real-world settings is a challenge for health professions educators. Our purpose in this article is to provide step-by-step instructions with worked examples for 5 absolute standard-setting methods that can be used to establish acceptable passing scores for performance examinations such as Objective Structured Clinical Examinations or standardized patient encounters. SUMMARY: All standards reflect the subjective opinions of experts. In this "how-to" article, we demonstrate procedures for systematically capturing these expert opinions using 5 research-based methods (Angoff, Ebel, Hofstee, Borderline Group, and Contrasting Groups). We discuss issues relating to selection of judges, use of performance data, and decision-making processes. CONCLUSIONS: Different standard-setting methods produce different passing scores; there is no "gold standard." The key to defensible standards lies in the choice of credible judges and in the use of a systematic approach to collecting their judgments. Ultimately, all standards are policy decisions. PMID- 16354142 TI - Comparison of aboriginal and nonaboriginal applicants for admissions on the Multiple Mini-Interview using aboriginal and nonaboriginal interviewers. AB - BACKGROUND: Achievement on grade point average and Medical College Admissions Test contribute as unintentional barriers to advancement of underrepresented minorities. So long as noncognitive measures mimic random number generators, they merely perpetuate such discrepancies. As reliable noncognitive measures are developed, it is crucial to ensure immunity from bias, enabling them to better dilute unintended discrimination of cognitive measures. PURPOSE: The Multiple Mini-Interview (MMI) is a recently developed, reliable (overall reliability = .70), noncognitive measure used for assessment of medical school applicants. Our purpose in this study was to evaluate whether any suggestion of bias existed in application of the MMI in its assessment of aboriginal medical school applicants. METHODS: In this study of the MMI (overall reliability = .70), each of 5 self declared aboriginal applicants and 7 general-pool applicants experienced the same 11 vetted interview stations with the same 6 aboriginal raters and 5 nonaboriginal raters. RESULTS: The Interviewer Type x Interviewee Type interaction was nonsignificant, p > .7. CONCLUSION: Based on the results of this study, it is recommended that MMI stations be vetted by aboriginally sensitive personnel, but neither aboriginal-specific rater training nor aboriginal rater assignment is required to ensure a level playing field for the assessment of applicants' personal qualities. PMID- 16354143 TI - Structured practice opportunities with a mnemonic affect medical student interviewing skills for intimate partner violence. AB - BACKGROUND: Low rates of partner violence inquiry and detection are reported in the medical setting. PURPOSE: To determine if a teaching module with a mnemonic improves interviewing skills. METHOD: Prospective randomized trial. A total of 43 medical students were assigned to either the intervention group (teaching module with guided discussion and practice highlighting use of a mnemonic) or the control group (general discussion and provision of the mnemonic at the end of the session). These students subsequently interviewed simulated patients. RESULTS: A total of 75% of the intervention group and 62% of the control group reported the mnemonic was helpful. A total of 68% of the intervention group and 45% of the control group asked a direct question about partner violence. Students who obtained a history of abuse consistently asked direct, nonjudgmental question(s). CONCLUSIONS: Students learn to perform desired interviewing skills more frequently when they have the benefit of guided discussion, practice, and memory aids. PMID- 16354144 TI - Abstracts from the Proceedings of the 2005 Annual Meetings of the Association for Surgical Education (ASE). PMID- 16354145 TI - Abstracts from the 2005 Annual Meeting of the Council on Medical Student Education in Pediatrics (COMSEP). PMID- 16354147 TI - Pimping perspectives: response to Wear. PMID- 16354157 TI - Uncoupled IP3 receptor can function as a Ca2+-leak channel: cell biological and pathological consequences. AB - Ca(2+) release via intracellular release channels, IP(3)Rs (inositol 1,4,5 trisphosphate receptors) and RyRs (ryanodine receptors), is perhaps the most ubiquitous and versatile cellular signalling mechanism, and is involved in a vast number of cellular processes. In addition to this classical release pathway there is limited, but yet persistent, information about less well-defined Ca(2+)-leak pathways that may play an important role in the control of the Ca(2+) load of the endo(sarco)plasmic reticulum. The mechanisms responsible for this 'basal' leak are not known, but recent data suggest that both IP(3)Rs and RyRs may also operate as Ca(2+)-leak channels, particularly in pathological conditions. Proteolytic cleavage or biochemical modification (such as hyperphosphorylation or nitrosylation), for example, occurring during conditions of cell stress or apoptosis, can functionally uncouple the cytoplasmic control domains from the channel domain of the receptor. Highly significant information has been obtained from studies of malfunctioning channels in various disorders; for example, RyRs in cardiac malfunction or genetic muscle diseases and IP(3)Rs in neurodegenerative diseases. In this review we aim to summarize the existing information about functionally uncoupled IP(3)R and RyR channels, and to discuss the concept that those channels can participate in Ca(2+)-leak pathways. PMID- 16354158 TI - Active cathepsins B, H, K, L and S in human inflammatory bronchoalveolar lavage fluids. AB - BACKGROUND INFORMATION: Chronic inflammation and tissue remodelling result from an imbalance between proteolytic enzymes and their inhibitors in the lungs in favour of proteolysis. While many studies have examined serine proteases (e.g. cathepsin G and neutrophil elastase) and matrix metalloproteases, little is known about the role of papain-like CPs (cysteine proteases). The present study focuses on the thiol-dependent cathepsins (CPs) and their specific cystatin-like inhibitors [CPIs (CP inhibitors)] in human inflammatory BALFs (BAL fluids, where BAL stands for broncho-alveolar lavage). RESULTS: Cathepsins B, K and S found were mostly zymogens, whereas cathepsins H and L were predominantly in their mature forms. Little immunoreactive cystatin C was found and the high- and low molecular-mass ('weight') kininogens were extensively degraded. The BALF procathepsins B and L could be activated autocatalytically, indicating that alveolar fluid pro-CPs are reservoirs of mature enzymes. Hydrolysis patterns of 7 amino-4-methylcoumarin-derived peptide substrates showed that extracellular alveolar CPs remain proteolytically active, and that cathepsins B and L are the most abundant thiol-dependent endoproteases. The CP/CPI balance was significantly tipped in favour of cathepsins (3- or 5-fold), as confirmed by the extensive CP dependent degradation of exogenous kininogens by BALFs. CONCLUSIONS: Although their importance for inflammation remains to be clarified, the presence of active cathepsins L, K and S suggests that they contribute to the extracellular breakdown of the extracellular matrix. PMID- 16354159 TI - Cytochrome P450 expression-induction profile and chemically mediated alterations of the WIF-B9 cell line. AB - BACKGROUND INFORMATION: WIF-B9 is a hybrid cell line obtained by fusion of rat hepatoma cells (Fao) and human fibroblasts (WI38). It exhibits the structural and functional characteristics of differentiated hepatocytes, including active bile canaliculi. The aim of the present study was to characterize the WIF-B9 cell line as a model for analysing drug-induced hepatic effects. The drug metabolism potential of WIF-B9 cells was identified by studying the rat and human CYP (cytochrome P450) mRNA constitutive expression profile and induction potential after exposure to reference inducers. The morphological alterations provoked by chemical entities were also characterized. RESULTS: Competitive reverse transcriptase-PCR revealed that four rat (1A1, 2B1/2, 2E1 and 4A1) and four human (1A1, 2Cs, 2D6 and 2E1) CYP mRNA isoforms were constitutively expressed in WIF-B9 cells. The rat CYP forms were expressed at levels 2-4 orders of magnitude higher than the human forms. Exposure for 20-72 h to increasing concentrations of CYP reference inducers (beta-naphthoflavone, 3-methyl cholanthrene, dexamethasone, phenobarbital, clofibrate and pregnenolone 16alpha-carbonitrile) revealed that the rat CYP 1A1, 1A2, 3A1, 3A2 and 4A1 and human CYP 1A1 and 2Cs mRNAs were inducible. Rat CYP 1A1 and 1A2 were the most inducible isoforms since they were overexpressed up to 100-fold after 20-48 h of treatment with beta-naphthoflavone. Human CYP 1A1 and 2Cs mRNAs were induced 3-fold after 48 h of treatment with phenobarbital. Other mechanisms involved in hepatotoxicity were explored using microscopy and immunofluorescence. The WIF-B9 cell line exhibited fragmentation and dilatation of bile canaliculi upon exposure to erythromycin, and to isoniazid and cytochalasins, respectively. Monensin promoted cell depolarization and cytoplasmic granulation. Ethionine promoted cytoplasmic vacuolation and dilatation of the Golgi structures. CONCLUSIONS: These results indicate that the CYP expression and induction profiles and the morphological features of WIF-B9 cells allow prediction in vitro of the induction and hepatotoxicity profiles of chemical entities. PMID- 16354160 TI - The potential role of caveolin-1 in inhibition of aquaporins during the AVD. AB - BACKGROUND INFORMATION: During apoptosis, the first morphological change is a distinct cell shrinkage known as the AVD (apoptotic volume decrease). This event is driven by a loss of intracellular K(+), which creates an osmotic gradient, drawing water out of the cell through AQPs (aquaporins). Loss of water in balance with K(+) would create a shrunken cell with an equivalent intracellular concentration of K(+) ([K(+)](i) = 140 mM). However, we have previously shown that the [K(+)](i) of the shrunken apoptotic cell is 35 mM, and this level is absolutely essential for the activation of apoptotic enzymes. We have recently found that AQPs are inactivated following the AVD, so that continued loss of K(+) will reduce the intracellular concentration to this critical level. Using thymocytes, we have investigated the expression profile and regulation of the AQP family members. RESULTS: In the present study, we have found that AQP1, AQP8 and AQP9 are present in non-apoptotic thymocytes and localized primarily to the plasma membrane. Expression and localization did not change when these cells were induced to undergo apoptosis by growth factor withdrawal for 24 h. To explore other possible mechanisms by which these water channels are inactivated, we investigated their association with CAV-1 (caveolin-1), binding to which is known to inactivate a variety of proteins. We found that CAV-1 is present in thymocytes and that this protein co-localizes with a portion of AQP1 in normal (non apoptotic) thymocytes. However, thymocytes induced to undergo apoptosis greatly increase their AQP1/CAV-1 association. CONCLUSIONS: Taken together, these results indicate that AQPs are localized to the plasma membrane of shrunken apoptotic thymocytes where increased binding to CAV-1 potentially inactivates them. AQP inactivation, coupled with continued K(+) efflux, then allows the [K(+)](i) to decrease to levels conducive for the activation of downstream apoptotic enzymes and the completion of the apoptotic cascade. PMID- 16354161 TI - Cytoskeletal and motor proteins facilitate trafficking of AQP1-containing vesicles in cholangiocytes. AB - BACKGROUND INFORMATION: We have previously showed that: (i) cholangiocytes contain AQP1 (aquaporin 1) water channels sequestered in intracellular vesicles; and (ii) upon stimulation with choleretic agonists such as secretin or dibutyryl cAMP (dbcAMP), the AQP1 vesicles move via microtubules to the apical cholangiocyte membrane to facilitate osmotically driven, passive water movement (i.e. ductal bile secretion). The aim of the present study was to determine which proteins and mechanisms regulate AQP1 trafficking in cholangiocytes. RESULTS: Using polarized cultured NMCs (normal mouse cholangiocytes) or NRCs (normal rat cholangiocytes) and affinity-purified antibodies, we performed immunofluorescent confocal microscopy on fixed cells or immunoblotting on cell lysates for actin, tubulin, kinesin and dynein, proteins known to regulate intracellular vesicle trafficking. By immunostaining, the appropriate orientation of the actin (i.e. sub-apical) and tubulin (i.e. generalized) cytoskeleton was apparent; kinesin and dynein displayed a homogeneous punctate distribution. Immunoblotting showed kinesin and dynein to be present in both cholangiocyte lysates and in isolated AQP1-containing vesicles. We utilized real-time fluorescence confocal microscopy of NMCs transfected with a GFP (green fluorescent protein)-AQP1 fusion construct in the presence and absence of dbcAMP. CONCLUSIONS: Our results provide additional insights into the potential molecular mechanisms of ductal bile secretion. PMID- 16354162 TI - Starvation-induced expression of autophagy-related genes in Arabidopsis. AB - BACKGROUND INFORMATION: Autophagy is a catabolic process for degradation of cytoplasmic components in the vacuolar apparatus. A genome-wide survey recently showed evolutionary conservation among autophagy genes in yeast, mammals and plants. To elucidate the molecular and subcellular machinery responsible for the sequestration and subsequent digestion of intracellular material in plants, we utilized a combination of morphological and molecular methods (confocal laser scanning microscopy, transmission electron microscopy and real-time PCR respectively). RESULTS: Autophagy in Arabidopsis thaliana suspension-cultured cells was induced by carbon starvation, which triggered an immediate arrest of cell growth together with a rapid degradation of cellular proteins. We followed the onset of these responses and, in this report, provide a clear functional classification for the highly polymorphic autophagosomes by which the cell sequesters and degrades a portion of its own cytoplasm. Quantification of autophagy-related structures shows that cells respond to the stress signal by a rapid and massive, but transient burst of autophagic activity, which adapts to the stress signal. We also monitored the real-time expressions of AtATG3, AtATG4a, AtATG4b, AtATG7 and AtATG8a-AtATG8i genes, which are orthologues of yeast genes involved in the Atg8 ubiquitination-like conjugation pathway and are linked to autophagosome formation. We show that these autophagy-related genes are transiently up-regulated in a co-ordinated manner at the onset of starvation. CONCLUSIONS: Sucrose starvation induces autophagy and up-regulates orthologues of the yeast Atg8 conjugation pathway genes in Arabidopsis cultured cells. The AtATG3, AtATG4a, AtATG4b, AtATG7 and AtATG8a-AtATG8i genes are expressed in successive waves that parallel the biochemical and cytological remodelling that takes place. These genes thus serve as early markers for autophagy in plants. PMID- 16354163 TI - Do Alix and ALG-2 really control endosomes for better or for worse? AB - Alix/AIP1 (ALG-2-interacting protein X/apoptosis-linked-gene-2-interacting protein 1) is an adaptor protein that was first described for its capacity to bind to the calcium-binding protein ALG-2 (apoptosis-linked gene 2), the expression of which seemed necessary for cell death. Over-expression of truncated forms of Alix blocks caspase-dependent and -independent mechanisms of cell death. Numerous observations in yeast and in mammalian cells suggest that Alix controls the making of and trafficking through endosomes called MVBs (multivesicular bodies), which are crucial intermediates within the endolysosomal system. In particular, deletion of Bro1, one of the yeast homologues of Alix, leads to an impairment in the function of MVBs, leading to mis-sorting of proteins normally destined to the vacuole. Mammalian Alix may have a similar function and has been shown to bind to lyso(bis)phosphatidic acid, ESCRT (endosomal sorting complex required for transport) proteins, endophilins and CIN85 (Cbl-interacting protein of 85 kDa), which are all main regulators of the endosomal system. EIAV (equine infectious anaemia virus) and HIV late domains use Alix to recruit the ESCRT machinery in order to bud from the cell surface, underscoring the crucial role of the protein in orchestrating membrane deformation. In this review I develop the hypothesis that the normal function of Alix in the endolysosomal system may be deviated by ALG-2 towards a destructive role during active cell death. PMID- 16354168 TI - Clinical snippets. PMID- 16354169 TI - Research snippets from the British Journal of Dermatology. 2005. PMID- 16354171 TI - Measuring the burden of atopic eczema in young children and the family unit. PMID- 16354172 TI - CD 200-mediated regulation of skin immunity. PMID- 16354173 TI - A merging duo in melanoma formation. PMID- 16354174 TI - The SNPs of RAF. PMID- 16354175 TI - Probing the immune dysregulation in cutaneous T cell lymphomas. PMID- 16354176 TI - Further evidence of multifunctionality in the keratinocyte: the endothelial protein C receptor. PMID- 16354177 TI - IVI g TREATMENT OF PEMPHIGUS: how it works and how to use it. PMID- 16354178 TI - In vitro differences between keratinocyte stem cells and transit-amplifying cells of the human hair follicle. AB - Epithelial stem cells within the human hair follicle are critical for hair development, hair cycling, wound healing, and tumorigenesis. We and others have previously shown that the hair follicle bulge area contains keratinocyte stem cells, whereas the hair matrix represents the proliferating and differentiating transit-amplifying (TA) cell compartment. In order to better characterize the phenotypic differences between human keratinocyte stem cells and their daughter TA cells, we compared the in vitro properties of cell adhesion, cell migration, clonogenicity, and in vitro life span. Epithelial outgrowths from the hair matrix appeared within 2 d of explant, whereas stem cell outgrowths appeared between 7 and 10 d after explant. Both populations form colonies; however, stem cells from telogen follicles formed more total colonies, and more colonies greater than 3 mm. Upon subculture, stem cells formed colonies until passage 6 and terminally differentiated at passage 7, whereas TA cells only formed colonies until passage 2. Stem cells express more beta1 integrin and adhere more rapidly to collagen IV. Most strikingly, TA cells showed a 7-fold greater mobility on migration assays than stem cells (0.704 vs 0.102 microm per min). These results help define the human hair follicle stem cell and TA cell phenotypes and correlate with the in vivo properties of these compartments. PMID- 16354179 TI - Development of the Childhood Atopic Dermatitis Impact Scale: initial validation of a quality-of-life measure for young children with atopic dermatitis and their families. AB - To measure the effects of atopic dermatitis (AD) on the quality of life of affected young children and their families, we developed a prototype 62-item instrument, the Childhood Atopic Dermatitis Impact Scale (CADIS). The prototype CADIS was developed from a comprehensive conceptual framework based on data from parents and clinicians. The instrument had eight subscales (four each for child and parent): physical health, emotional health, physical functioning, and social functioning. The goal of this work was to test the validity of and to refine the prototype of CADIS. Two hundred seventy parents of children under the age of 6 y with AD responded to the instrument. Content validity was demonstrated by expert and parent reviews of the drafted and refined instrument, and by analyzing parents' responses to open-ended questions about their children's skin disease. Construct validity was assessed in exploratory factor analyses which supported a refinement in the conceptual framework to consist of two dimensions with five domains: child dimensions (symptoms and activity limitation/behavior), and parent dimensions (family/social function, sleep, and emotions). Seventeen items were eliminated, yielding a 45-item refined version of CADIS (score 0-180) with evidence of content and construct validity and suggested use in clinical research. PMID- 16354180 TI - Molecular mechanisms of junctional epidermolysis bullosa: Col 15 domain mutations decrease the thermal stability of collagen XVII. AB - Mutations in the collagen XVII gene, COL17A1, are associated with junctional epidermolysis bullosa. Most COL17A1 mutations lead to a premature termination codon (PTC), whereas only a few mutations result in amino acid substitutions or deletions. We describe here two novel glycine substitutions, G609D and G612R, and a splice site mutation resulting in a deletion of three Gly-X-Y amino acid triplets. In order to investigate the molecular pathomechanisms of non-PTC mutations, G609D and G612R and two previously known substitutions, G627V and G633, and deletion of the amino acids 779-787 were introduced into recombinant collagen XVII. The thermal stability of the mutated collagens was assessed using trypsin digestions at incremental temperatures. All the four glycine substitutions significantly destabilized the ectodomain of collagen XVII, which manifested as 16 degrees C-20 degrees C lower T(m) (midpoint of the helix-to-coil transition). These results were supported by secondary structure predictions, which suggested interruptions of the collagenous triple helix within the largest collagenous domain, Col15. In contrast, deletion of the three full Gly-X-Y triplets, amino acids 779-787, had no overall effect on the stability of the ectodomain, as the deletion was in register with the triplet structure and also generated compensatory changes in the NC15 domain. PMID- 16354181 TI - Genetic and environmental influences on skin pattern deterioration. AB - Sun exposure has been known to cause histological changes in the dermal layer of the skin. Using deterioration in the fine reticular patterning of the epidermal stratum corneum (skin pattern, as measured on the Beagley-Gibson scale) as a proxy measure of histological changes in the dermal layer, previous studies have typically assumed that degradation of skin pattern is largely caused by sun exposure. A twin study comprising 332 monozygotic twin pairs and 488 dizygotic twin pairs at ages 12, 14, and 16 was used to investigate the etiology of variation in skin pattern, particularly in relation to measured sun exposure and skin color. Our results indicate that although self-reported sun exposure is a significant contributor to variation in skin pattern, its effect is small, explaining only 3.4% of variation in skin pattern at age 14. Additive genetic effects explain 86% of variation in skin pattern at age 12 but these effects reduce with age so that 75% of variation is due to additive genetic effects at age 14 and 72% at age 16. This trend of diminishing genetic influences continues into adulthood, with 62% of variation due to non-additive genetic factors in a smaller adult sample (aged 32-86). Skin color explains 10.4% of variation in skin pattern at age 12, which is due to additive genetic influences common to both. Melanin content appears to provide a protective effect against skin pattern deterioration, perhaps because of the structural differences in melanosomes between different skin types or the free radical scavenging properties of melanin. PMID- 16354182 TI - Characterization of CD 200-receptor expression in the murine epidermis. AB - CD 200 is a widely expressed transmembrane glycoprotein that transmits an inhibitory signal after ligation of the structurally homologous CD 200-receptor-1 (CD 200 R1). Recently, we showed that CD 200 is expressed on keratinocytes and plays a role in protecting hair follicles from autoimmune attack. Here, we report the characterization of cell surface and mRNA expression of CD 200 R1 by cells of the murine epidermis. In addition, we report mRNA expression for other members of the CD 200 R-family (R2-R4) by quantitative real-time RT-PCR. Variable levels of CD 200 R1, R2, R3, and R4 mRNA were detected in bulk epidermal cell suspensions. Freshly isolated Langerhans cells (LC) preferentially expressed CD 200 R1. Consistent with an inhibitory role for CD 200:CD 200 R1 interaction, LC obtained from mice deficient in CD 200 (CD 200(-/-)) were in a heightened state of activation as compared with wild-type (CD 200(+/+)) cells. Freshly isolated dendritic epidermal T cells (DETC) expressed low levels of CD 200 R1, R2, and R3 mRNA, but they preferentially increased cell surface and mRNA expression of CD 200 R1 upon activation in vitro. In functional assays using sub-optimal CD3 signaling, immobilized CD 200 inhibited DETC proliferation and cytokine secretion. Collectively, these results suggest that CD 200:CD 200 R interactions may play a role in regulating both LC and DETC in cutaneous immune reactions. PMID- 16354183 TI - Immunology of the human nail apparatus: the nail matrix is a site of relative immune privilege. AB - The nail apparatus is constantly exposed to environmental damage. It requires effective immune responses to combat infection, while avoiding the loss of nail production and regeneration by autoaggressive immunity. By immunohistology, we define here previously unknown characteristics of the normal human nail immune system (NIS). Compared with other regions of nail epithelium, human leukocyte antigen (HLA)-A/B/C expression is prominently down regulated on both keratinocytes and melanocytes of the proximal nail matrix (PNM), whereas HLA-G(+) is upregulated here. Together with the expression of macrophage migration inhibitory factor in PNM, this may serve to inhibit an natural killer (NK) cell attack on major histocompatibility complex (MHC) class Ia-negative PNM. PNM also displays strong immunoreactivity for potent, locally generated immunosuppressants such as transforming growth factor-beta1, alpha-melanocyte stimulating hormone, insulin-like growth factor-1, and adrenocorticotropic hormone, exhibits unusually few CD1a(+), CD4(+), or CD8(+), NK, and mast cells. Finally, MHC class II and CD 209 expression on CD1a(+) cells in and around the PNM is reduced, indicating diminished antigen-presenting capacity. Thus, the NIS strikingly differs from the skin immune system, but shows intriguing similarities to the hair follicle immune system, including the establishment of an area of relative immune privilege in the PNM. This nail immune privilege may offer a relative safeguard against autoimmunity. But, the localized intraepithelial defect of innate and adaptive immunity in the PNM revealed here also may impede effective anti-infection defense. PMID- 16354184 TI - Differential expression of genes involved in skin homing, proliferation, and apoptosis in CD4+ T cells of patients with atopic dermatitis. AB - CD4+ T cells play a critical role in allergic diseases, both in the affected tissue as well as systemically. Our objective was to investigate the in vivo activation state of peripheral blood CD4+ T cells of atopic dermatitis (AD) patients by analyzing gene expression profiles of unstimulated CD4+ T cells. mRNA samples from blood CD4+ T cells, isolated from five AD patients and seven healthy controls (HC), were analyzed using oligonucleotide arrays. Differentially regulated genes were validated by quantitative PCR (Q-PCR) in a larger group of patients with AD, in a group of patients with allergic asthma (AA), and HC subjects. In addition, "typical" T helper type 1 (Th1)- and Th2-related genes were analyzed by Q-PCR. Microarray analysis revealed differential expression of 52 genes in AD patients. Q-PCR confirmed several differentially regulated genes in AD, including CCR10, CRTH2, C-JUN, and NR4A2. Two groups of genes with highly correlating gene expression levels involved in tissue homing and proliferation or apoptosis, respectively, were identified. No marked differences were found in gene expression levels of typical Th1 or Th2 genes in AD or in AA patients. This study demonstrates that peripheral blood, unstimulated CD4+ T cells in AD patients show differentially expressed genes involved in tissue homing, proliferation, and apoptosis. No marked expression differences of "typical" atopy genes were found. PMID- 16354185 TI - Hyperproduction of IFN-gamma by CpG oligodeoxynucleotide-induced exacerbation of atopic dermatitis-like skin lesion in some NC/Nga mice. AB - Under conventional conditions, NC/Nga mice spontaneously develop an atopic dermatitis (AD)-like skin lesion accompanied by immunoglobulin E (IgE) hyperproduction and the expression of T helper 2 (Th2) cytokines. CpG DNA activates a strong interferon-gamma (IFN-gamma)-dominated T helper 1 (Th1) response, while inhibiting Th2-dependent allergies. In this study, we examined whether CpG oligodeoxynucleotide (ODN) could prevent the development of the skin lesions in NC/Nga mice. Sixteen of 26 NC/Nga mice did not exhibit dermatitis after CpG ODN was administered intraperitoneally every 2 wk for a total of five times. CpG ODN administration induced IFN-gamma production, which inhibited the production of Th2 cytokines (interleukin (IL)-4, IL-5, and IL-13) in both spleen and lymph node cells and culminated in a decrease in the serum IgE level. These data suggest that the CpG ODN has a therapeutic effect against AD; however, some mice (10 of 26) treated with CpG ODN exhibited an exacerbation of dermatitis accompanied by the hyperproduction of IFN-gamma, although Th2 cytokines were suppressed. These results suggest that the suppression of Th2 cytokines may not completely prevent dermatitis and that IFN-gamma may play a role in developing dermatitis in some NC/Nga mice. PMID- 16354186 TI - High expression levels of keratinocyte antimicrobial proteins in psoriasis compared with atopic dermatitis. AB - Recently, it was shown that lesional skin of atopic dermatitis patients expresses low levels of some antimicrobial peptides, compared with psoriasis patients. Here we performed microarray analysis on mRNA from purified lesional epidermal cells of patients with chronic plaque psoriasis and chronic atopic dermatitis, to investigate whether this is a general phenomenon for host defense proteins, and how specific it is for this class of molecules. Microarray data were confirmed on a selected set of genes by quantitative PCR and at the protein level by immunohistochemistry. We found overexpression of many antimicrobial proteins in keratinocytes from psoriatic skin compared with atopic dermatitis skin. Interestingly, we observed that markers of normal differentiation and the activated/hyperproliferative epidermal phenotype were expressed at equal levels. Chronic lesions of psoriasis and atopic dermatitis patients are remarkably similar with respect to cellular proliferation. We conclude that psoriatic epidermis expresses high levels of host defense proteins compared with atopic dermatitis epidermis, and this phenomenon appears to be specific for these proteins. It remains to be investigated whether this is caused by genetic polymorphisms in pathways leading to an epidermal antimicrobial response, or by differences in the cellular infiltrate in psoriasis compared with atopic dermatitis. PMID- 16354187 TI - Differential expression of fatty acid transport proteins in epidermis and skin appendages. AB - Epidermis and sebocyte-derived lipids are derived both from de novo synthesis and through uptake of fatty acids from the circulation. Plasma membrane proteins can significantly contribute to the latter process. In particular, fatty acid transport proteins (FATP/solute carrier family 27) are integral transmembrane proteins that enhance the uptake of long-chain fatty acids into cells. Using specific antisera against all six mammalian FATP, we found that both human and mouse skin express FATP1, -3, -4, and -6. In adult skin, FATP1 and -3 are expressed predominantly by keratinocytes, whereas FATP4 is strongly expressed by sebaceous glands and FATP6 by hair follicle epithelia. Sustained barrier disruption leads to increases in FATP1 and -6 levels as well as a robust increase in CD36 protein. Notably, expression of FATP1 by embryonic keratinocytes at day 18.5 was lower, and FATP4 increased in comparison with adult epidermis. Together, these findings indicate that FATP are not only expressed by different cell types within the skin, but also that their localization is dynamically regulated during development. PMID- 16354188 TI - Quantification of human tissue kallikreins in the stratum corneum: dependence on age and gender. AB - Human tissue kallikreins are a family of 15 trypsin or chymotrypsin-like secreted serine proteases (hK1-hK15). hK5, hK6, hK7, hK8, and hK13 have been identified in the stratum corneum (SC), stratum granulosum, and skin appendages. It has been reported that hK5 and hK7 degrade desmosomes/corneodesmosomes, suggesting that kallikreins are responsible for desquamation. We report the quantification of hK5, hK6, hK7, hK8, hK10, hK11, hK13, and hK14 in the SC by ELISA and their variation among age groups. The total SC trypsin and chymotrypsin-like activities were also measured. The amount of hK7, hK8, and hK11 (ng per mg dry weight) were high, and varied from 6 to 14, hK5 (2.0-4.0) was present at intermediate levels, and hK10 (0.65-1.0), hK14 (0.1-0.3), hK6 (0.1-0.3), and hK13 (0.02-0.1) were present at lower levels. hK6 and hK14 were significantly lower in females between 20 and 59 y. hK5, hK7, hK10, hK11, and hK14 were not significantly different across the age groups. hK8 was lowest at extremes of age (highest at 30-39 y), hK6 was lower at >30 y, and hK13 was lower at >20 y. Overall trypsin-like activity did not differ across age groups but was higher in subjects <11 y. Overall chymotrypsin-like activity was not related to age. In conclusion, we found multiple kallikreins in the SC and suggest that these enzymes may be responsible for desquamation through an enzymatic cascade pathway. PMID- 16354189 TI - Keratinocyte growth factor promotes melanosome transfer to keratinocytes. AB - Melanogenesis and melanosome transfer from the melanocytes to the neighboring keratinocytes are induced by ultraviolet radiation and modulated by autocrine and paracrine factors. Keratinocyte growth factor (KGF/fibroblast growth factor (FGF)7) is a paracrine mediator of human keratinocyte growth and differentiation. We evaluated the influence of KGF on melanosome transfer in co-cultures of keratinocytes and melanocytes. Immunofluorescence analysis using anti-tyrosinase and anti-human cytokeratin antibodies, phagocytic assays using fluorescent latex beads, and ultrastructural analysis indicated that KGF is able to induce melanosome transfer acting only on the recipient keratinocytes and as a consequence of a general role of KGF in the promotion of the phagocytic process. Inhibition of proteinase-activated receptor-2, to block the Rho-dependent phagocytic pathway, or of the Src family tyrosine kinases, to inhibit the Rac dependent pathway, showed that KGF promotes phagocytosis through both mechanisms. Increased expression of the KGF receptor (KGFR) on the keratinocytes by transfection led to increased phagocytosis of latex beads following KGF treatment, suggesting that the KGF effect is directly mediated by KGFR expression and activation. Moreover, confocal microscopic analysis revealed that KGFR localize in phagosomes during KGF-induced phagocytosis, suggesting a direct role of the receptor in regulating both the early steps of uptake and the intracellular traffic of the phagosomes. PMID- 16354190 TI - Iron in skin of mice with three etiologies of systemic iron overload. AB - In human hemochromatosis, tissue toxicity is a function of tissue iron levels. Despite reports of skin toxicity in hemochromatosis, little is known about iron levels in skin of individuals with systemic iron overload. We measured skin iron and studied skin histology in three mouse models of systemic iron overload: mice with a deletion of the hemochromatosis (Hfe) gene, mice fed a high iron diet, and mice given parenteral injections of iron. In Hfe(-/-) mice, iron content in the epidermis and dermis was unexpectedly the same as in Hfe(+/+) mice, and there were no histological abnormalities detected after 30 wk. A high iron diet produced increased iron in the epidermis of both normal and Hfe(-/-) animals; a high diet increased iron in the dermis only in Hfe(-/-) mice. Increased skin iron was not associated with other histological changes, even after 19 wk. Parenteral administration of iron produced increased iron in the epidermis and dermis, and gave the skin a bronze hue. These results show that the amount and distribution of iron in the skin depends on the etiology of iron overload. It appears that neither Hfe deletion nor elevated skin iron alone can account for cutaneous manifestations reportedly seen in humans with hereditary hemochromatosis. PMID- 16354191 TI - Topical liver x receptor activators accelerate postnatal acidification of stratum corneum and improve function in the neonate. AB - In neonatal rat stratum corneum (SC), pH declines from pH 6.8 at birth to adult levels (pH 5.0-5.5) over 5-6 d. Liver X receptor (LXR) activators stimulate keratinocyte differentiation, improve permeability barrier homeostasis, and accelerate the in utero development of the SC. In this manuscript we determined the effect of LXR activators on SC acidification in the neonatal period and whether these activators correct the functional abnormalities in permeability barrier homeostasis and SC integrity/cohesion. Formation of the acid SC-buffer system was accelerated by topically applying the LXR activator, 22(R) hydroxycholesterol, and non-oxysterol activators of LXR, TO-901317, and GW-3965. A sterol which does not activate LXR had no effect. LXR activation increased secretory phospholipase A(2) (sPLA(2)) activity and conversely, inhibition of sPLA(2) activity prevented the LXR induced increase in SC acidification, suggesting that increasing sPLA(2) accounts in part, for the LXR stimulation of acidification. LXR activation resulted in an improvement in permeability barrier homeostasis, associated with an increased maturation of lamellar membranes attributable to an increased beta-glucocerebrosidase activity. SC integrity cohesion also normalized in LXR-activator-treated animals and was associated with an increase in corneodesmosomes and in desmoglein 1 expression. These results demonstrate that LXR activators stimulate the formation of an acidic SC and improve both permeability barrier homeostasis and SC integrity/cohesion. PMID- 16354192 TI - Different consequences of beta1 integrin deletion in neonatal and adult mouse epidermis reveal a context-dependent role of integrins in regulating proliferation, differentiation, and intercellular communication. AB - There are conflicting reports of the consequences of deleting beta1 integrins from the epidermis of transgenic mice. Epidermal thinning with normal differentiation and lack of inflammation has been observed; conversely, epidermal thickening, abnormal differentiation, and dermal fibrosis can occur. beta1 integrin deletion results in decreased epidermal proliferation, yet on wounding the proliferative defect is overcome. To distinguish primary from secondary consequences of beta1 integrin loss, we compared epidermal beta1 deletion at E14.5 via K5Cre and 4-hydroxy-tamoxifen induced deletion in adulthood via K14CreER. As reported previously, there was dermo-epidermal splitting, inflammation, reduced proliferation, and hair follicle and sebaceous gland loss in 30-d-old K5Cre beta1-null mice. These changes were not observed 30 d after beta1 integrin deletion in adult epidermis, however, and there were no changes in the hair follicle stem cell compartment. Deletion in adult epidermis revealed a previously unreported correlation between the level of beta1 integrins and proliferation in the interfollicular epidermis that was remarkably consistent with human epidermis. In addition, the number of melanocytes in interfollicular epidermis was greatly increased. Our results highlight the context-dependent effects of beta1 integrin deletion and suggest that inflammation may be responsible for some of the K5Cre beta1-null phenotype. PMID- 16354193 TI - Disturbed epidermal structure in mice with temporally controlled fatp4 deficiency. AB - So far, little is known about the physiological role of fatty acid transport protein 4 (Fatp4, Slc27a4). Mice with a targeted disruption of the Fatp4 gene display features of a human neonatally lethal restrictive dermopathy with a hyperproliferative hyperkeratosis, a disturbed epidermal barrier, a flat dermal epidermal junction, a reduced number of pilo-sebaceous structures, and a compact dermis, demonstrating that Fatp4 is necessary for the formation of the epidermal barrier. Because Fatp4 is widely expressed, it is unclear whether intrinsic Fatp4 deficiency in the epidermis alone can cause changes in the epidermal structure or whether the abnormalities observed are secondary to the loss of Fatp4 in other organs. To evaluate the functional role of Fatp4 in the skin, we generated a mouse line with Fatp4 deficiency inducible in the epidermis. Mice with epidermal keratinocyte-specific Fatp4 deficiency developed a hyperproliferative hyperkeratosis with a disturbed epidermal barrier. These changes resemble the histological abnormalities in the epidermis of newborn mice with total Fatp4 deficiency. We conclude that Fatp4 in epidermal keratinocytes is essential for the maintenance of a normal epidermal structure. PMID- 16354194 TI - Biological effects of SLURP-1 on human keratinocytes. AB - A novel paradigm of keratinocyte (KC) regulation via nicotinic acetylcholine receptors (nAChR) has been discovered in studies of SLURP (secreted mammalian Ly 6/urokinase-type plasminogen activator receptor-related protein)-1 in Mal de Meleda. We cloned human SLURP-1 and produced recombinant protein and the monoclonal antibody 336H12-1A3 that visualized native SLURP-1. SLURP-1 ligated the conventional ligand-binding site of KC nAChR, showing a higher affinity to the [(3)H]nicotine-, compared with the [(3)H]epibatidine-sensitive nAChR. SLURP-1 significantly (p<0.05) increased the activities of caspases 3 and 8, and the number of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells. The pro-apoptotic activity of SLURP-1 exceeded that of tumor necrosis factor-alpha, suggesting the involvement of separate pathways. In a series of real-time PCR and in-cell western experiments, SLURP-1 significantly (p<0.05) upregulated expression of transglutaminase type I cytokeratin 10, p21, and caspase-3. In the presence of the agonist carbachol, the effects of SLURP-1 on gene expression were augmented, which is in keeping with the notion that SLURP 1 acts as an allosteric agonist at the KC nAChR. Thus, the changes in the cell state induced by SLURP-1 could result from nAChR-mediated effects on the KC gene expression. These results suggest that the biological role of SLURP-1 in the epidermis is to provide fine tuning of the physiologic regulation of KC functions through the cholinergic pathways. PMID- 16354195 TI - Recurrent patterns of dual RB and p53 pathway inactivation in melanoma. AB - Two growth inhibitory hurdles that must be overcome by the evolving cancer cell include pathways regulated by RB and p53. In human melanoma cells, inactivation of a single locus, CDKN2A, can lead to abrogation of both RB and p53 functionality through loss of the two CDKN2A cognate transcripts-p16 and p14ARF, respectively. We thus set out to assess how recurrent patterns of genetic injury at three critical human melanoma loci-CDKN2A, TP53, and CDK4-cooperate to disrupt both RB and p53 pathways. Overall, 77.8% of the melanoma cell lines analyzed showed genetic evidence of dual RB and p53 pathway compromise; this percentage is even higher if protein expression loss is considered. Although homozygous deletion of all three critical CDKN2A exons (exons 1 beta, 1 alpha, and 2) represent the most common mechanism, concurrent loss of CDKN2A(Exon1 alpha) and CDKN2A(Exon1 beta) and simultaneous point mutagenesis of CDK4 and TP53 reflect alternative cassettes of dual inactivation. In cell lines with isolated CDKN2A(Exon2) mutations, coincident alterations in TP53 or deletion of CDKN2A(Exon1 beta) suggest that p16 transcript may be preferentially targeted over the p14ARF transcript as additional p53 pathway lesions are recruited. Moreover, predictive modeling of CDKN2A(Exon2) missense mutations further suggests that the amino acid substitutions in this region negatively impact p16 to a greater extent than p14ARF. Taken together, our data point to a clear need in human melanoma cell lines, as in its murine counterpart, to disrupt both RB and p53 pathways and recurrent mechanisms may play into the unique genetic vulnerabilities of this tumor type. PMID- 16354196 TI - BRAF polymorphisms and risk of melanocytic neoplasia. AB - Somatic mutations of the BRAF gene are common in melanomas and nevi but the contribution of polymorphisms in this gene to melanoma or nevus susceptibility remains unclear. An Australian melanoma case-control sample was typed for 16 single nucleotide polymorphisms (SNP) within the BRAF gene, and five SNP in three neighboring genes. The sample comprised 755 melanoma cases from 740 families stratified by family history of melanoma and controls from 635 unselected twin families (2,239 individuals). Ancestry of the cases and controls was recorded, and the twins had undergone skin examination to assess total body nevus count, degree of freckling, and pigmentation phenotype. Genotyping was carried out via primer extension followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry. SNP in the BRAF gene were found to be weakly associated with melanoma status but not with development of nevi or freckles. The estimated proportion of attributable risk of melanoma due to variants in BRAF is 1.6%. This study shows that BRAF polymorphisms predispose to melanoma but the causal variant has yet to be determined. The burden of disease associated with this variant is greater than that associated with the major melanoma susceptibility locus CDKN2A, which has an estimated attributable risk of 0.2%. PMID- 16354197 TI - Changes in different melanocyte populations during hair follicle involution (catagen). AB - Melanin synthesis in the hair follicle (HF) is strictly coupled to the growth stage of the hair cycle and is interrupted during follicle regression (catagen) and resting. Using tyrosine-related protein 2 (Trp)2-LacZ transgenic mice as a model, we show that distinct melanocyte subpopulations of the HF display distinct patterns of apoptosis and survival during catagen. Melanocytes located in the outer root sheath express Bcl-2 and are TUNEL-negative. Part of the pigment producing melanocytes located above the follicular papilla expresses Fas, TUNEL, and is likely to undergo apoptosis, whereas the other part of these melanocytes expresses c-kit, Bcl-2, and becomes visible in the follicular papilla. During late catagen, TUNEL and Ki-67 negative melanocytes expressing Bcl-2 are seen in the secondary germ of the HF. Lack of proliferation in the follicular melanocytes during catagen suggests that secondary hair germ of late catagen HF is most likely repopulated by melanocytes arising from the outer root sheath or follicular papilla of early/mid-catagen HF. Taken together, these data suggest a possible scenario and mechanisms of the remodeling of the follicular pigmentary unit during HF anagen-catagen-telogen transition and may be used for the establishing in vivo models for pharmacological modulation of melanocyte apoptosis and survival during the hair cycle. PMID- 16354198 TI - Noxious heat and scratching decrease histamine-induced itch and skin blood flow. AB - The aim of this study was to assess the effect of thermal stimuli or distal scratching on skin blood flow and histamine-induced itch in healthy volunteers. Twenty-one healthy volunteers participated in the study. Baseline measurements of skin blood flow were obtained on the flexor aspect of the forearm. These measurements were compared with skin blood flow after various stimuli: heating the skin, cooling the skin, noxious cold 2 degrees C, noxious heat 49 degrees C, and scratching via a brush with controlled pressure. Afterwards histamine iontophoresis was performed and skin blood flow and itch intensity were measured immediately after the above-mentioned stimuli. Scratching reduced mean histamine induced skin blood flow and itch intensity. Noxious heat pain increased basal skin blood flow but reduced histamine-induced maximal skin blood flow and itch intensity. Cold pain and cooling reduced itch intensity, but neither affected histamine-induced skin blood flow. Sub-noxious warming the skin did not affect the skin blood flow or itch intensity. These findings suggest that heat pain and scratching may inhibit itch through a neurogenic mechanism that also affects skin blood flow. PMID- 16354199 TI - Circulating natural killer lymphocytes are potential cytotoxic effectors against autologous malignant cells in sezary syndrome patients. AB - Patients with advanced cutaneous T cell lymphoma (CTCL) exhibit profound defects in cell-mediated immunity. Although it has been suggested that Sezary syndrome (SS) patients have a decreased natural killer (NK) lymphocyte activity, nothing has been reported concerning the sensitivity of Sezary cells to NK lymphocyte mediated cytotoxicity. Peripheral blood NK cells from healthy donors were tested against Sezary tumoral cell lines as well as against freshly isolated Sezary cells. Further, we studied their ability to exhibit antibody -dependent cell mediated cytotoxicity using either the murine anti-CD158k/KIR3DL2 monoclonal antibody (moAb) AZ158 that specifically recognizes Sezary cells, or the anti-CD52 monoclonal antibody alemtuzumab. The results show that Sezary cell lines are susceptible to NK lymphocyte lysis. More importantly, we found that freshly isolated malignant cells are killed either by IL-2 activated allogeneic NK lymphocytes or when the tumor lymphocyte targets are incubated with an anti-MHC class I F(ab)'2 antibody. Further, anti-KIR3DL2 and anti-CD52 moAb can enhance the NK lysis. Finally, we report that NK lymphocytes isolated from SS patients are potentially cytotoxic lymphocytes against autologous malignant Sezary cells. These findings indicate that antitumor-mediated NK lymphocyte cytotoxic activity can be triggered in patients with CTCL and raise the possibility of developing novel therapeutic strategies by stimulating their innate immunity. PMID- 16354200 TI - Endothelial protein C receptor and protease-activated receptor-1 mediate induction of a wound-healing phenotype in human keratinocytes by activated protein C. AB - Activated protein C (APC) is a natural anticoagulant and inhibitor of inflammation that can stimulate keratinocyte wound repair in vitro and promote wound healing in vivo. The signaling mechanisms, however, are unknown and a keratinocyte receptor for APC has not been identified. Here, we show that cultured human keratinocytes from neonatal foreskins express the endothelial protein C receptor (EPCR). EPCR was also strongly expressed by lower epidermal layers of neonatal foreskin as determined by immunohistochemistry. In cultured keratinocytes, EPCR expression was upregulated by the addition of APC and inhibited by tumor necrosis factor-alpha. Addition of APC stimulated cell proliferation, production of matrix metalloproteinase-2, activation of ERK and p38 kinase signaling pathways, and expression of protease-activated receptor (PAR)-1. A monoclonal antibody, RCR252, which blocks APC binding to EPCR, or a blocking antibody to PAR-1, abolished APC's effects on keratinocytes. In summary, this study demonstrates that EPCR, a major receptor of protein C pathway, is expressed by human keratinocytes, and facilitates APC's function on keratinocytes via activation of PAR-1 pathway. Our findings highlight a possible new role for the protein C pathway in skin physiology and help elucidate the mechanisms of action by which APC promotes wound healing. PMID- 16354201 TI - Microarray analysis of gene expression in cultured skin substitutes compared with native human skin. AB - Cultured skin substitutes (CSS), prepared using keratinocytes, fibroblasts, and biopolymers, can facilitate closure of massive burn wounds by increasing the availability of autologous tissue for grafting. But because they contain only two cell types, skin substitutes cannot replace all of the functions of native human skin. To better understand the physiological and molecular differences between CSS and native skin, we undertook a comprehensive analysis of gene expression in native skin, cultured keratinocytes, cultured fibroblasts, and skin substitutes using Affymetrix gene chip microarrays. Hierarchical tree clustering identified six major clusters of coordinately regulated genes, using a list of 1030 genes that were the most differentially expressed between groups. These clusters correspond to biomarker pools representing expression signatures for native skin, fibroblasts, keratinocytes, and cultured skin. The expression analysis revealed that entire clusters of genes were either up- or downregulated upon combination of fibroblasts and keratinocytes in cultured skin grafts. Further, several categories of genes were overexpressed in CSS compared with native skin, including genes associated with hyperproliferative skin or activated keratinocytes. The observed pattern of expression indicates that CSS in vitro, which display a well-differentiated epidermal layer, exhibit a hyperproliferative phenotype similar to wounded native skin. PMID- 16354202 TI - Functional significance of Smad2 in regulating basal keratinocyte migration during wound healing. AB - Members of the transforming growth factor-beta (TGF-beta) superfamily are critical regulators for wound healing. Transduction of TGF-beta signaling depends on activation of Smad2 and Smad3 by heteromeric complexes of ligand-specific receptors. Mice lacking Smad3 show accelerated wound healing, whereas the biological significance of Smad2-mediated TGF-beta signaling in wound healing remains unknown. To understand the function of Smad2 in regulating wound healing, we investigated the effect of Smad2 overexpression on epithelialization of incision wounds. Cutaneous wounds made in K14-Smad2 mice showed delayed healing. This delay in wound healing resulted from a defect in basal keratinocyte migration in K14-Smad2 mice. Instead of basal keratinocytes, the suprabasal layer of keratinocytes migrated into the wound region. Furthermore, overexpression of Smad2 activated the Smad2/Smad4 complex in keratinocytes and inhibited keratin 16 (K16) expression. As K16 functions as a critical mediator for reorganization of keratin filaments following skin injury, we propose that altered K16 expression affects the migration of basal keratinocytes in the K14-Smad2 mice. Taken together, these findings demonstrate a crucial role of TGF-beta signaling mediator Smad2 in regulating keratinocyte migration and re-epithelialization during wound healing. The K14-Smad2 transgenic mice can serve as an animal model for the investigation of TGF-beta signaling mechanism in regulating wound healing. PMID- 16354203 TI - Nevus distribution in a Utah melanoma kindred with a temperature-sensitive CDKN2A mutation. PMID- 16354204 TI - Infrared-mediated hyperthermia is effective in the treatment of scleroderma associated Raynaud's phenomenon. PMID- 16354205 TI - Selective cyclooxygenase-2 inhibition produces heterogeneous erythema response to ultraviolet irradiation. PMID- 16354210 TI - Clinical course of congenital nephrotic syndrome and Denys-Drash syndrome in Japan. AB - BACKGROUND: The prognosis of Japanese patients with congenital nephrotic syndrome (CNS) and Denys-Drash syndrome (DDS) is not clear. METHODS: Five patients with CNS and four patients with DDS, which causes secondary CNS, were studied retrospectively. RESULTS: Seven patients were sporadic and two DDS patients were identical twins. Five CNS patients presented with edema within 3 months of birth. In four DDS patients, edema was not noted and end-stage renal disease developed between 7 months and 6 years of age. Of these five CNS patients, one patient had cerebral thrombosis and cytomegalovirus pneumonia at the onset and another patient died during dialysis. Frequent intravenous albumin administration required, growth and development during infancy were varied. Of the nine patients with CNS and DDS, seven received renal transplantation and were alive with functioning grafts at the last follow up. Catch-up growth was observed in five patients after transplantation. Five school-aged patients attended school and received adequate grades and two adults worked full-time. Of the DDS patients, dysuria due to hypospadias persisted in one patient and treatment for hypogonadism was needed in one patient. CONCLUSIONS: CNS and DDS were diagnosed early after onset and adequate treatment was started. Growth and development after renal transplantation were relatively good. Thrombotic episodes or severe infection in CNS patients was difficult to manage and complications resulting from DDS affected the quality of life. PMID- 16354211 TI - Familial cases of Henoch-Schonlein purpura in eight families. AB - BACKGROUND: Familial cases of Henoch-Schonlein purpura (HSP) have rarely been reported. METHODS: Familial cases of HSP were reviewed by medical records of 418 children with HSP. RESULTS: Two members developed HSP in eight families. HSP occurred in a mother and her daughter in one family and in siblings, including one pair of twin sisters, in seven other families. Four pairs of patients developed HSP at the same age. Three pairs presented HSP within 1 month of each other and the other pairs presented HSP between 9 months and 5 years. Seven patients had a history of allergic diseases. The clinical courses of 12 patients were reviewed. Upper respiratory tract infection preceded HSP in 10 patients, two of whom had elevated antistreptolysin-O titers. No pairs of patients in a family received the same drugs before the onset of HSP. Abdominal pain was noted in eight patients, arthralgia in six and nephritis in four. Severity of skin lesions, presence of abdominal pain and nephritis, and serum IgA levels at the acute stage varied among family members of HSP. CONCLUSIONS: The incidence of HSP in family members of children with HSP seems to be high. Onset at the same age and onset of HSP within 1 month in siblings have not previously been reported. There were no characteristic or similar findings between two patients of the same family. No trigger or genetic factor causing HSP was identified. PMID- 16354212 TI - Different patterns of cytokines, ECP and immunoglobulin profiles at two adverse drug reactions in a patient. AB - OBJECTIVES: Drug-induced hypersensitivity syndrome (HS) is a rare but life threatening disease. We experienced carbamazepine-induced HS in a 14-year-old boy, who had cefaclor-induced cutaneous eruptions 15 months later. To clarify the mechanisms of HS and the differences between two diseases we studied this case in detail. METHODS: We investigated the associated viral agents by polymerase chain reaction and the specific antibodies. We also studied the mechanism of diseases by measuring chemical mediators including cytokines, ECP and immunoglobulins. RESULTS: The patient was diagnosed as having carbamazepine-induced HS associated with reactivation of human herpesvirus 6 based on the clinical course and laboratory data including drug-induced lymphocyte stimulation tests. Similarly, the diagnosis of cefaclor-induced eruption without any viral reactivation was made. Serum levels of IFN-gamma, IL-6, TNF-alpha, IL-5 and ECP were increased significantly at HS but mildly at cefaclor-induced eruptions. Furthermore, we detected transient hypogammaglobulinemia only at HS. CONCLUSIONS: This is the first report of anticonvulsant-induced HS followed by antibiotic-induced eruptions in a patient. In addition, we demonstrated difference in serum levels of inflammatory cytokines, immunoglobulins, activated eosinophils and viral reactivation between these diseases. This case would contribute to the understanding of the pathophysiology of adverse drug reactions including HS. PMID- 16354213 TI - Effects of montelukast on symptoms and eNO in children with mild to moderate asthma. AB - BACKGROUND: Asthma is a chronic inflammatory airway disease. Exhaled nitric oxide (eNO) is a marker reflecting airway inflammation. This study was conducted to investigate whether montelukast, a leukotriene receptor antagonist, could be used for the management of asthma and how fast the montelukast sodium decreased airway inflammation as demonstrated by eNO levels. METHODS: Twenty children aged 6-14 years (mean age: 9.2 +/- 2.4 years; mean weight 30 +/- 4.6 kg) with mild to moderate asthma were recruited for the study. They received montelukast plus an inhaled short-acting beta2 agonist as open and uncontrolled therapy. Asthma score (AS) and peak expiratory flow rate (PEFR) and eNO concentrations were measured at pretreatment (0 week) and post-treatment (1 and 2 weeks) as well as 2 weeks after withdrawal of therapy. RESULTS: In one week, the eNO levels (33.3 +/- 15.5 p.p.b. vs 14.8 +/- 8.6 p.p.b.; P < 0.05), and AS (4.2 +/- 1.3 vs 1.8 +/- 1.3; P < 0.05) decreased rapidly, and PEFR (206.9 +/- 69.7 L/min vs 236.2 +/- 69.8 L/min; P < 0.05) increased. Concurrent beta2 agonist use decreased from a mean +/- SD of 2.2 +/- 0.4-1.3 +/- 0.3 puffs per weeks (P < 0.05). After the withdrawal of treatment for 2 weeks, the eNO levels (29.2 +/- 16.1 p.p.b) rebounded again, although the improvements in AS (1.1 +/- 1.3) and PEFR (245.0 +/- 91.3 L/min) persisted. CONCLUSION: Oral montelukast sodium treatment of these children with mild to moderate asthma effectively improved asthmatic symptoms and suppressed airway inflammation in 1 week, suggesting that this leukotriene antagonist combined with short-acting beta2 agonists may provide effective treatment option in mild to moderate childhood asthma. Larger, controlled, and double-blinded studies are needed to confirm these preliminary open uncontrolled observations. PMID- 16354214 TI - Sodium cromoglycate therapy in wheezing infants: preliminary evidence of beneficial outcome at early school age. AB - BACKGROUND: In order to affect the natural course of childhood wheezing and asthma, anti-inflammatory therapy is often prescribed for young wheezing children, but there is lack of long-term follow-up data. METHODS: Eighty-two of the original 100 children, hospitalized for wheezing under the age of 2 years in 1992-1993, were re-examined at school age in 1999. The children had participated in an open, randomized, parallel-group trial including a 4-month intervention with inhaled sodium cromoglycate (SCG) or budesonide (BUD). The baseline data, including data on atopy, eosinophilia and viral etiology, were prospectively collected on admission. RESULTS: At early school age (median 7.2 years), asthma was present in 33 (40%) children. There was less asthma in the original SCG (21%) than in the control group (54%) (OR 0.23; 95% CI 0.07-0.77). The figure was 46% in the BUD group. When the analyses were performed separately for atopic and non atopic infants, the difference was significant only among atopics. The lowered risk for asthma in the SCG group remained significant in the multivariate logistic regression analysis when adjusted for age, sex and atopy, and further when adjusted for earlier episodes of wheezing and respiratory syncytial virus identification. However, after adjustment for blood eosinophilia, the significance was lost, albeit the risk for asthma remained low (OR 0.21; 95% CI 0.04-1.12). A sensitivity analysis, which was done by including the six drop-outs of the SCG group as unfavorable and the 12 drop-outs of other groups as favorable outcomes in the model, did not change the direction of the result (OR 0.70; 95% CI 0.26-1.89). CONCLUSIONS: An early SCG intervention in infants hospitalized for wheezing was associated with a lowered risk for early school-age asthma, especially in infants with evidence of atopy. PMID- 16354215 TI - Decreased total antioxidant capacity and increased oxidative stress in passive smoker infants and their mothers. AB - BACKGROUND: Smoking has many adverse health effects in infants and adults. The purpose of the study was to study the effect of passive cigarette smoking on oxidative and antioxidative status of plasma in passive smoker infants and their mothers and to compare with those of non-smokers. METHODS: Subjects were randomly chosen from infants aged 8-26 weeks and their mothers aged 20-34 years. Passive smoker infants (n = 29) and their mothers (n = 29) were defined as having other family members who smoked six or more cigarettes per day continually for at least 8 weeks. Non-smokers were defined as infants (n = 30) and their mothers (n = 24) who had never been exposed to passive smoking. The antioxidative status of plasma were perused by measuring the total antioxidant capacity. Oxidative status was evaluated by predicating total peroxide level, oxidative stress index, protein oxidation and lipid peroxidation. RESULTS: Plasma concentrations of total antioxidant capacity were significantly lower in passive smoker infants and their mothers than non-passive smoker infants and their mothers. However, lipid peroxidation and oxidative stress index were remarkably higher in passive smoker infants and their mothers than those of non-passive smoker infants and their mothers. There were significant correlations between the oxidative and antioxidative parameters of the passive smoker infants and their mothers. CONCLUSIONS: Oxidants are increased and antioxidants are decreased in passive smoker infants and their mothers than those of non-smokers. Passive smoker infants and their mothers are exposed to potent oxidative stress. PMID- 16354216 TI - Familial Mediterranean fever in a childhood population in eastern Turkey. AB - BACKGROUND: Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent attacks of inflammation of serosal membranes. Amyloidosis is the most severe complication of the disease The aim of this study was to explore the magnitude of the FMF problem and to describe clinical phenotypic and genotypic profile in the childhood population in Eastern Turkey. METHODS: In this study, 52 patients who were diagnosed as FMF between January 2000 and January 2003 in Department of Pediatrics, Ataturk University Hospital, were included. The diagnosis of FMF was based on typical clinical and laboratory features. The 12 FMF mutations were investigated in the patients. RESULTS: Of the 52 patients, 30 (57.7%) were girls, 22 (42.3%) were boys, and the age ranged from 9 months to 15 years (8.5 +/- 3.2 years). A positive family history for FMF was noted in 33 (63.5%) patients. The mean onset age was 6 +/- 3.4 (from 8 months to 14 years). Nineteen children (36.5%) were symptomatic below the age of 5 years. Abdominal pain was observed in 50 (96.2%), fever in 42 (80.8%), arthralgia in 29 (55.8%), arthritis in 18 (34.6%), splenomegaly in 11 (21.2), hepatomegaly in 15 (28.8%), myalgia in 11 (26.2%), erysipelas-like erythema in 10 (19.2%), thoracic pain in four (7.7%), protracted febrile myalgia in three (5.8%), and seizures in two (3.8%). The most frequent mutation was the M694V/M694V. Clinical presentation of the patients was not different in respect with genotypes (P > 0.05). Two patients had chronic renal disease suggestive of amyloidosis. CONCLUSION: It was noted that the FMF patients in this study had a broad spectrum of mutation combination, which might reflect the intercultural interactions of ancient ethnic groups that lived in Anatolia, and these mutations were not significantly different in respect to clinical presentations. PMID- 16354217 TI - Diurnal variation of height in children. AB - BACKGROUND: Diurnal variation of height is largely ignored in the assessment of growth in children. Diurnal decrease in stature may effect the reliability of height measurement. In this study, the degree of differences in daily height measurement were evaluated. METHODS: A total of 478 children, aged 3-15 years (mean age 9.9 +/- 2.3 years) were enrolled in the study. The height of children were measured twice in a day. The first measurements were taken between 09:00 and 10:00 h and the second measurements between 15:00 and 16:00 h. RESULTS: Children's height measurements differences were +1.8 to -2.7 cm, and a mean of 0.47 +/- 0.05 cm decrease was determined. CONCLUSIONS: Little differences of height measurement could be very important in evaluating the short child. When the result of height measurement is recorded, recording the time of day is offered. PMID- 16354218 TI - Vitamin D status of mothers and their neonates in Kuwait. AB - BACKGROUND: There has been a resurgence of nutritional rickets in children in many developing countries and some of the developed countries. Children between 6 and 18 months old are commonly affected. In order to find out the association between vitamin D and rickets we studied the vitamin D status of the neonates and their mothers in Kuwait. METHODS: Two hundred and fourteen full-term pregnant mothers and their neonates were selected from two hospitals in Kuwait. All mothers had normal vaginal delivery. On the day of delivery 2.5 mL of maternal blood and 2.5 mL of cord blood samples were withdrawn. Serum 25-hydroxyvitamin D (25OHD) was determined in duplicate by radioimmunoassay using an Incstar kit. Quality control analyses were done using several between and within run experiments. RESULTS: A total of 128 mother-neonate pairs were selected from the Al-Adan hospital and 86 from the Maternity Hospital. The mean age and parity of the mothers were similar in both hospitals. The mean (+/- SD) 25OHD levels of the mothers and the neonates in the Adan hospital were 13.3 (6.5) ng/mL and 8.2 (6.5) ng/mL, respectively. The corresponding values in the Maternity Hospital were 17.6 (12.4) and 8.1 (7.3) ng/mL for the mothers and the neonate, respectively. Serum 25OHD of the mothers and their newborn infants were highly correlated (r = 0.790, P < 0.001). CONCLUSIONS: Results demonstrate that 40% of the mothers and 60% of the neonates are vitamin D deficient on the day of delivery. The vitamin D of the mothers and neonates are highly correlated (r = 0.790, P < 0.001). PMID- 16354219 TI - Primary malignant skin tumors in children: etiology, treatment and prognosis. AB - OBJECTIVE: The aim of the study was to evaluate the etiology, treatment and prognosis of the malignant skin tumors in children. METHODS: Twenty-one patients who had been diagnosed with malignant skin tumors between 1972 and 2003 were retrospectively analyzed. Age range was 0.5-20 years (median 9), and the male/female ratio was 12/9. We had nine (42.9%) patients with malignant melanoma, five (23.8%) with primary skin non-Hodgkin lymphoma, three (14.3%) with Kaposi sarcoma (KS), two (9.5%) with basal cell carcinoma (BCC), and two (9.5%) with squamous cell carcinoma (SCC). RESULTS: We could define the etiologic factors in only nine (42.9%) patients. Two KS cases were associated with renal transplantation, two cases of malignant melanoma occurred within the area of giant hairy cell nevus, one melanoma patient previously had bone marrow transplantation due to Gricelli syndrome, one patient with BCC had xeroderma pigmentosum and the other BCC had got radiotherapy due to previous diagnosis of medulloblastoma. One SCC patient also had xeroderma pigmentosum and the other had previous skin burn. Overall survival rate was 77%. Melanoma patients were treated successfully with high-dose interferon. CONCLUSION: Although malignant skin tumors are rare in childhood, the prognosis is relatively better than it is for adults. Malignant melanoma was the most frequent tumor. Forty-three per cent of our patients had an underlying defect in their immune barriers which is thought to be responsible for the development of their malignancies. PMID- 16354220 TI - Oxidative stress in infants born to preeclamptic mothers. AB - BACKGROUND: Oxidative stress may play an important role in the pathophysiology of preeclampsia. An increase in lipid peroxidation products and a decrease in antioxidant activity in preeclamptic women have been reported in many papers. The objective of this study was to evaluate oxidative stress in infants born to preeclamptic mothers. METHODS: Malondialdehyde (MDA) and glutathione (GSH) levels and glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were measured in cord plasma of infants born to preeclamptic (n = 18) or normotensive (n = 9) mothers. RESULTS: Gestational age was similar in both groups. The mean birth weight was significantly lower in the preeclamptic group (P = 0.007). Maternal age, primigravidity, antenatal steroid use, premature rupture of the membranes, clinical chorioamnionitis and adverse neonatal outcomes including sepsis, respiratory distress syndrome and neonatal mortality did not differ between groups. Cesarean delivery was significantly higher in the preeclamptic group. There was no significant difference in cord plasma levels of MDA and GSH, and activity of GPx between the preeclamptic and control groups. SOD was found to be increased in preeclamptic group (P = 0.03). CONCLUSIONS: We concluded that although cord plasma MDA levels were similar in both the preeclamptic and control groups, increased SOD activity might be an indicator of increased oxidative stress in infants born to preeclamptic mothers. PMID- 16354221 TI - Foreign body aspiration in children. AB - BACKGROUND: The aim was to investigate the role of physical and radiological findings before bronchoscopy in the diagnosis of foreign body aspiration (FBA). METHODS: We retrospectively reviewed the clinical records for 82 patients (mean age 26.4 +/- 21.4 months, range 9 months to 13.5 years; 49 males) with a history suggestive of foreign body aspiration. RESULTS: The presence of a foreign body in the airways was confirmed in 70 children (85.4%) (mean age 25 +/- 14.1 months, 45 boys). Of the 70 children, 63 patients (90%) were under 3 years of age, with a peak incidence during the second year. Of the 70 foreign bodies retrieved, 46 (60%) were vegetable and 35 (76%) of these were nuts. In 42% of the patients the foreign body was located in the right bronchial tree. The most frequent physical findings observed in our patients were persistent cough (75%), localized decreased breath sound (62.8%) and localized wheezing (30%). The clinical triad (concomitant cough, localized wheezing and decreased breath sound) was present in 11 patients (15.7%). All clinical findings had a high positive predictive value with poor sensitivity. In 11 patients (20%) chest X-rays were normal. Five foreign bodies (9.1%) were radiopaque. The most frequent radiological findings observed were localized air trapping (43.6%), followed by atelectasis (40%). The diagnostic sensitivity was 80% and the specificity 33% for the presence of a single positive radiological finding. CONCLUSIONS: Our study confirmed that clinical symptoms and radiological findings before bronchoscopy have a low diagnostic value in children with a history of FBA. PMID- 16354222 TI - Clinical characteristics of children with snakebite poisoning and management of complications in the pediatric intensive care unit. AB - BACKGROUND: Venomous snakebite is an emergency condition with high morbidity and mortality in childhood. Nearly all venomous snakes in Turkey are members of the Viperidae family and show poisonous local and hematotoxic effects. METHODS: A total of 77 children (mean age 9.9 +/- 2.9 years; age range 3-14 years) with venomous snakebites were investigated. General characteristics of the children, species of the snakes, localization of the bite, clinical and laboratory findings, treatment approaches, complications and prognosis were evaluated. RESULTS: The male to female ratio was 1.4. Ninety-one per cent of cases were from rural areas. Most of the bites were seen in May and June. Mean duration between snakebites and admissions to our department was 13 +/- 6.5 h. According to a clinical grading score, 57.1% of patients presented to us as grade II. Mean leukocyte count, aspartate aminotransferase, lactate dehydrogenase, creatinine phosphokinase and protrombin time levels were above the normal ranges and mean activated partial tromboplastin time was below the normal range. Platelet counts inversely correlated with the grading score and duration of hospitalization. The most common complication that occurred during the treatment was tissue necrosis (13%). The mean hospital stay time was 6.3 +/- 6 days. Three children with disseminated intravascular coagulation died. Fasciotomies were performed to seven (9.1%) children due to compartment syndrome. Of 10 children with tissue necrosis, three (3.9%) had finger amputation and seven (9.1%) had toe amputation. Higher grading score on admission, platelet count below 120 000/mm3, AST over 50 IU/L and existence of evident ecchymosis were found as significant risk factors for development of serious complications by logistic regression analysis. CONCLUSIONS: Snakebite poisoning is an emergency medical condition that is particularly important in childhood. The envenomations are still considerable public health problems with a high morbidity and mortality in rural areas of Turkey. PMID- 16354223 TI - The efficacy of two different lipid-based amphotericin B in neonatal Candida septicemia. AB - BACKGROUND: Fungal sepsis is becoming more frequent in neonatal intensive care units (NICU) and has a high mortality rate due to the invasive nature of the disease and to the insufficiency of low doses and high incidence of renal problems with effective doses of amphotericin B. New generation lipid formulated amphotericin B preparations may be more efficient because they are less toxic to be applied in target doses. However, there is limited experience in neonates and preterm infants. METHODS: The charts of 917 patients admitted to NICU between 2001 and 2003 were reviewed and the data of 21 patients with systemic Candida infection, requiring different amphotericin B therapy, were analyzed. RESULTS: Infants with fungal septicemia were treated with amphotericin B lipid complex (Abelcet)(n = 10) and liposomal amphotericin B (AmBisome)(n = 9) for a mean duration of 21 and 18 days. The mean gestational age of the patients was 30.9 +/- 4.2 weeks and mean birth weight was 1536 +/- 714 g. Two patients in the Abelcet group and one patient in the AmBisome group died during therapy. Fungal eradication was achieved in 16 surviving infants and mean eradication time was 8.1 +/- 2.6 days and mean duration of therapy was 19.2 +/- 4.1 days. Mortality rates related to treatment failure were similar being 20% in the Abelcet group and 11% in the AmBisome group. No patient showed severe side-effects from the antifungal therapy; the incidence of minimal side-effects were similar in both groups and they were elevated serum transaminase levels in six patients, increased serum creatinine in one patient and hypokalemia in one patient. CONCLUSION: Both preparations have the same benefits for the treatment of neonatal fungal sepsis and they can be used safely in neonates including very low birth weight infants. However, the clinician must keep in mind the cost of treatment. PMID- 16354224 TI - Allogenic bone marrow transplantation for late-infantile neuronal ceroid lipofuscinosis. PMID- 16354225 TI - A molecular lesion in a Japanese patient with severe phenotype of 3 methylglutaconic aciduria type I. PMID- 16354226 TI - A novel mutation of the PHKA2 gene in a patient with X-linked liver glycogenosis type 1. PMID- 16354227 TI - The Jewish-type Cohen syndrome complicated with refractory bronchial asthma and depression. PMID- 16354228 TI - Hemorrhagic bullous lesions in Henoch-Schonlein purpura. PMID- 16354229 TI - Interrupted aortic arch type C associated with DiGeorge syndrome in 22q11.2 deletion: first case detected in Japan. PMID- 16354230 TI - Laparoscopic gastropexy for chronic gastric volvulus complicated by pathologic aerophagia in a boy. PMID- 16354231 TI - Forehead swelling caused by Pott's puffy tumor in a 9-year-old boy with sinusitis. PMID- 16354232 TI - Celiac disease with Takayasu arteritis. PMID- 16354233 TI - Guidelines for diagnosis and management of cardiovascular sequelae in Kawasaki disease. PMID- 16354234 TI - Acute renal failure in patients with pre-existing renal dysfunction following coronary artery bypass grafting. AB - BACKGROUND: Pre-existing renal dysfunction predisposes to acute renal failure (ARF) in patients undergoing coronary artery bypass grafting. We assessed the incidence and impact of the development of ARF in this patient population in our unit. METHODS: One-hundred and six patients had a preoperative serum creatinine of >or=0.13 mmol/L and underwent coronary artery bypass grafting in the year 2000. The incidence of ARF (as defined by a >or=50% rise in postoperative serum creatinine), hospitalization days, dialysis requirement, in-hospital and 1-year mortality, and potential risk factors for ARF were recorded. RESULTS: Of the patients recorded, 43/104 (41.35%) developed ARF following coronary artery bypass grafting. Patients with ARF stayed in hospital longer (P < 0.02). Ten out of forty-three patients required some form of dialysis and the in-hospital mortality of the renal failure group was 23% compared to 3.1% in the other group (P < 0.002). One year postoperatively, the group with renal failure had significantly worse survival (71.8% vs 98%P < 0.0001). CONCLUSION: For patients undergoing coronary artery bypass grafting, pre-existing renal dysfunction predisposes to the development of ARF, this is associated with prolonged hospitalization and increased mortality. PMID- 16354235 TI - Alterations in the biochemical markers of renal function after sevoflurane anaesthesia. AB - AIM: This study has been carried out to see whether renal function is acutely altered in patients undergoing sevoflurane anaesthesia. For this purpose, the urinary levels of markers of renal tubular function, namely leucine amino peptidase (LAP), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and beta-2 microglobulin (beta-2M), and urinary albumin as a predictor of renal glomerular function were measured before and after sevoflurane anaesthesia. METHODS: This study was comprised of 20 patients (11 males and nine females) aged 18-55, who underwent various elective surgical procedures under general anaesthesia. Urine samples of all patients were collected before and 1, 2 and 8 h after the anaesthesia. The levels of LAP, GGT, beta-2M, and albumin were then expressed as factored by urinary creatinine. In all patients, the anaesthesia was maintained with sevoflurane (2% end-tidal) at a high flow-rate (6 L/min). RESULTS: Urinary beta-2M and LAP levels after anaesthesia were unchanged (P > 0.05). While urinary GGT and ALP levels were found elevated in the first hour, LDH levels were higher in the second hour (P < 0.05). They returned to normal levels in the later periods after the anaesthesia. Urinary albumin excretion (UAE) was significantly elevated in the second hour after the anaesthesia (P < 0.001). Although UAE was decreased in the eighth hour after the anaesthesia, it still remained higher than the pre-anaesthesia level (P < 0.001). CONCLUSIONS: These results suggest that a 2% end-tidal concentration of sevoflurane at a high flow-rate (6 L/min) acutely alters renal glomerular function but does not have a significant acute effect on biochemical markers of renal tubular damage. PMID- 16354236 TI - Wasp envenomation-induced acute renal failure: a report of three cases. AB - Acute renal failure is an unusual complication of wasp stings. We report three cases who developed acute renal failure after multiple wasp stings (Vespa magnifica). Two patients had evidence of intravascular haemolysis and rhabdomyolysis whereas one patient investigation showed no evidence of intravascular haemolysis or rhabdomyolysis. All three cases had impaired liver functions. Oligo-anuria was seen in all three of the patients and all of them required dialytic support. One patient died of massive gastrointestinal bleeding while the remaining two recovered completely. Although acute renal failure after wasp stings is typically caused by acute tubular necrosis in the setting of haemolysis or rhabdomyolysis, in some patients, renal failure may result from a direct nephrotoxic effect or acute interstitial nephritis from a hypersensitivity reaction to the wasp venom. PMID- 16354237 TI - Late onset of familial nephrotic syndrome associated with a compound heterozygous mutation of the podocin-encoding gene. AB - A case of two young adult brothers with nephrotic syndrome secondary to focal segmental glomerulosclerosis is reported. Steroid resistance prompted us to perform genetic studies. These showed a compound heterozygous mutation of NPHS2, the gene encoding podocin. It was composed of a missense mutation in exon 7 (A284V) and the non-neutral polymorphism R229Q in exon 5. We review literature supporting the genetic basis of the disease. PMID- 16354238 TI - Nocturnal haemodialysis: an Australian cost comparison with conventional satellite haemodialysis. AB - Dialysis is an expensive therapy, particularly considering its recurrent, protracted nature while patient numbers are also increasing. To afford dialysis for those in need, smarter, more efficient use of limited funds is mandatory. Newer techniques and improved equipment now permit safe, highly effective haemodialysis (HD) at home, alone and while asleep. Indeed, the increase in treatment hours and frequency achieved through nocturnal HD both increase HD efficiency and reduce cardiovascular stress when comparing nocturnal HD (6 nights/week for 8 h/treatment) to conventional daytime HD (4 h/treatment, three times/week). This study compares the expenditure of two distinct HD programmes in the same renal service during the Australian financial year 2003/2004. A conventional satellite HD unit (SHDU) and a nocturnal home HD programme (NHHD(6)) are compared, with both programmes 'notionalised' to 30 patients. The state derived funding models under which these programmes operate are explained. All wage costs, recurrent expenditure, fixed costs and the estimated costs of building and infrastructure are included. The total NHHD(6) programme expenditure was 33,392 Australian dollars/patient per year (103.82 Australian dollars/treatment) and was 3,892 Australian dollars/patient per year less (a 10.75% saving) when compared with the SHDU expenditure of 36,284 Australian dollars/patient per year (232.58 Australian dollars/treatment). This represented an annual 116,750 Australian dollars programme saving for a 30 patient cohort. Potential additional NHHD(6) savings in erythropoietin, hospitalization and social security dependence were also identified. Home-based therapies are clinically sound, effective and fiscally prudent and efficient. Funding models should reward home-based HD. Health services should encourage home training and support systems, sustaining patients at home wherever possible. PMID- 16354239 TI - Peritoneal clearance of homocysteine with icodextrin or standard glucose solution exchange. AB - BACKGROUND: The aim of the study was to assess plasma homocysteine concentration in peritoneal dialysis patients, and to compare the effect of different peritoneal solutions (glucose-based and icodextrin-based) on peritoneal clearance of homocysteine. METHODS: The study group comprised 10 chronic peritoneal dialysis patients; the control group comprised 15 healthy, age-matched non-obese subjects with normal renal function. Patients with vitamin B(12) or folate deficiency were excluded. In all subjects, plasma homocysteine and dialysis adequacy parameters were assessed at baseline. The clearance study was carried out with 2.27% glucose and 7.5% icodextrin solutions (12-h dwell time). RESULTS: Mean dialysate concentration of homocysteine was similar for both glucose and icodextrin solutions (8.3 +/- 3.2 and 8.4 +/- 1.9 micromol/L, respectively), but homocysteine clearance was significantly higher for icodextrin than glucose solution (1.82 +/- 0.57 vs 1.39 +/- 0.53 mL/min per 1.73 m(2)P = 0.01). Net ultrafiltration after icodextrin solution was also higher than after glucose solution (599 +/- 136 mL vs 134 +/- 337 mL, P < 0.01). A correlation between total plasma level of homocysteine and its peritoneal clearance was found (r = 0.69; P = 0.03). CONCLUSION: It appears that peritoneal elimination of homocysteine depends primarily on its plasma concentration. Icodextrin-based solution for peritoneal dialysis seems to be more efficient in homocysteine elimination than a standard glucose-based solution. PMID- 16354240 TI - Elevated serum interleukin-18 levels might reflect the high risk of hospitalization in patients on peritoneal dialysis. AB - BACKGROUND: Interleukin (IL)-18 is a potent pro-inflammatory cytokine and plays a central role in atherosclerotic plaque rupture and accelerates atherosclerosis. AIM: The aim of this study was to determine serum IL-18 levels in patients on peritoneal dialysis (PD) and to assess their relationship with hospitalization. METHODS: Forty-three PD patients and 20 healthy individuals were enrolled in this study. We investigated the relationship of the serum concentrations of IL-18 and other well-established atherosclerotic markers, such as asymmetric dimethylarginine (ADMA). Hospitalization data from over a 18-month period were prospectively obtained on all 43 PD patients. Classic factors were entered into a Cox regression model to predict first hospitalization. RESULTS: The serum levels of IL-18 in patients on PD were significantly higher than those of healthy individuals (228.5 +/- 140.3 pg/mL vs 154.8 +/- 44.7 pg/mL, P < 0.05, respectively). Furthermore, serum IL-18 levels showed a positive correlation with duration of PD, serum beta2 microglobulin and serum ADMA levels. Mean serum levels of IL-18 were significantly higher among patients who had experienced at least one hospitalization than those who had not (279.9 +/- 164.3 vs 158.5 +/- 43.9 pg/mL, P = 0.0426). Furthermore, the relative risk for first hospitalization for each increase in IL-18 (pg/mL) levels was associated with a 1.182 (95% confidence interval, 1.012-1.364; P = 0.0071) increase in the risk for future hospitalization events. CONCLUSION: The present study suggests the elevated serum IL-18 levels might increase the risk for future hospitalization in patients on PD. PMID- 16354241 TI - Melatonin corrects reticuloendothelial blockade and iron status in haemodialysed patients. AB - AIM: Treatment of anaemia in haemodialysed patients in the setting of inflammation usually displays high levels of serum ferritin (>800 ng/mL) and low transferrin saturation (TSAT) (<20%) despite i.v. iron supplementation, thus proving iron trapping in the reticuloendothelial system. Melatonin has been reported to reduce cytokine production and, in dialysis patients, to prevent oxidative stress resulting from iron and erythropoietin treatment. METHOD: In this study, we evaluated a group of 10 patients undergoing haemodialysis who displayed elevated serum ferritin (981 +/- 44.6 ng/mL) and TSAT <20% (15.6 +/- 3.8%) after having received 1.2 g of i.v. iron dextran over a period of 8 weeks. These patients received oral melatonin, 6 mg/day at night for 30 days. RESULTS: After this treatment, all of them markedly increased TSAT values, reaching 35.5 +/- 6.7% (P < 0.0001 vs basal values). In addition, ferritin values decreased to 754.4 +/- 263.7 ng/mL (P < 0.05), and serum iron dramatically increased in all of the patients under study (42.4 +/- 9.4 vs 109.7 +/- 24.3 microg/dL; P < 0.0001). Values for haematocrit (28.6 +/- 2.7 vs 31.9 +/- 3.57%; P < 0.05) and haemoglobin (9.19 +/- 0.97 vs 10.04 +/- 1.29 g/dL; P < 0.05) were also improved. Measurements were then repeated 2 weeks after melatonin withdrawal, showing an impressive decrease in TSAT (16.4 +/- 5.3%; P < 0.00001) and serum iron (48 +/- 14.7 microg/dL; P < 0.0001) values and an almost significant increase in ferritin values (954.4 +/- 86 ng/mL; P < 0.054). CONCLUSION: The present study demonstrates that melatonin may strongly correct the reticuloendothelial blockade seen in dialysis patients under an inflammatory status, thus allowing a better management of iron derangements and renal anaemia. PMID- 16354242 TI - Propylthiouracil attenuates acetaminophen-induced renal damage in the rat. AB - BACKGROUND: To date, there is no specific antidote to acetaminophen poisoning. Propylthiouracil (PTU) has been shown to be protective against acetaminophen (APAP)-induced liver damage in rats; however, the nephroprotective effect of propylthiouracil has not been studied yet. METHODS: In order to verify this, rats were given different doses of PTU (100, 200 or 400 mg/kg per body weight, orally) 1 h before a nephrotoxic dose of APAP (1,000 mg/kg per body weight, intraperitoneally (i.p.)). RESULTS: Propylthiouracil pretreatment significantly reduced APAP-induced nephrotoxicity in a dose-dependant manner, as evidenced by reduction in plasma creatinine and by amelioration of renal pathology (interstitial congestion, tubular cell degeneration and necrosis). CONCLUSION: The mechanism of protection by PTU is probably not due to the sparing effect of non-protein thiol (approximately 95% of which is reduced glutathione), as similar depletion of renal glutathione was observed regardless of PTU pretreatment; other mechanisms are suggested. PMID- 16354243 TI - Multiple myeloma-associated cast nephropathy with crystal structure: case report and review of the literature. AB - The causes of renal failure are diverse. Among them, monoclonal gammopathy is one important but easily-missed cause in aged people. Monoclonal gammopathy may produce a large number of abnormal immunoglobulins and/or fragments and produce different kinds of deposition in tissues, including cast, crystal, fibril and granules. Cast nephropathy is considered the hallmark of the renal disease in patients with multiple myeloma. Crystaglobulinemia syndrome and crystal nephropathy, on the other hand, have been rarely reported. Herein, we report a case of multiple myeloma presented with irreversible renal failure. The biopsy showed massive crystal deposition in bone marrow and kidneys. PMID- 16354244 TI - Insulin resistance: more important to identify than quantify. PMID- 16354245 TI - Should we quantify insulin resistance in patients with renal disease? AB - Cardiovascular disease is a major cause of morbidity and mortality in dialysis patients. Vascular disease develops before the initiation of dialysis, and it is now recognized that chronic kidney disease (CKD) is an independent risk factor for cardiovascular disease. Death from cardiovascular disease is a more common endpoint of CKD than progression to dialysis. There are multiple mechanisms that contribute to the increased vascular risk of CKD, one of which is the presence of insulin resistance (IR). CKD is characterised by many features of the metabolic syndrome, and features of IR are also observed in dialysis and transplant patients. IR may be quantified by several different methods. One such method is homeostatic model assessment (HOMA) technique, which derives a measurement of IR from fasting plasma glucose and insulin concentrations. The HOMA index has been demonstrated to be an independent predictor of survival in dialysis patients. CKD is characterised by a chronic inflammatory response and abnormalities in the production and regulation of adipose tissue derived proteins, which may contribute to the development of IR. There are a range of interventions including diet and exercise programmes or medications that may influence IR; however, the impact of these interventions in the context of CKD has not been systematically evaluated. PMID- 16354246 TI - Sirolimus: its role in nephrology. AB - Sirolimus (Rapamycin, Wyeth Pharmaceuticals Australia Pty Ltd, Baulkham Hills, NSW, Australia) (SRL) has received increasing attention as an immunosuppressant in renal and other solid organ transplantation. Sirolimus is the first marketed agent in a new class of drugs with a novel mechanism of action. Sirolimus binds, like tacrolimus, to a member of the FK binding protein (FKBP) family. The SRL/FKBP complex binds to the protein kinase mTOR. Binding to mTOR blocks activation of signal transduction pathways causing arrest of the cell cycle in the G1 phase. It is now known that mTOR is a central regulator of cell growth and proliferation. The immunosuppressive properties of SRL are due primarily to blockade of interleukin-2 (IL-2)-induced proliferation of T cells. There is still much to be learnt about how best to use the drug. The key advantage over the current choice of immunosuppressive agents is the ability to preserve renal function and pathology while producing excellent rejection-free, graft survival rates. Thus, SRL may find its pivotal role as a calcineurin inhibitors replacement in patients whose grafts are affected by chronic allograft nephropathy. A second major driver for use may prove to be the impact of SRL on cancer incidence and prognosis. Studies still need to be performed to evaluate the best timing for commencement of SRL and the optimal dosage to minimize side effects. PMID- 16354247 TI - A study of gene expression of cyclophilin in childhood refractory nephrotic syndrome and its clinical significance. AB - AIM: The purpose of this study was to investigate the clinical significance of gene expression of cyclophilin (CyP) in children with refractory nephrotic syndrome (NS). METHODS: The levels of leukocyte mRNA expression in 33 cases of childhood refractory NS were determined by reverse transcription polymerase chain reaction (RT-PCR). The CyP levels in these children were compared with those of a healthy control groups. RESULTS: Cyclophilin (408 bp) and beta-actin bands (234 bp) were distinct on electrophoresis. The levels of leukocyte CyP mRNA expression were higher in children with mesangio-proliferative glomerulonephritis (MsPGN) and focal segmental glomerulosclerosis (FSGS) than in minimal-change nephrotic syndrome (MCNS), whereas in membrano-proliferative glomerulonephritis (MPGN) they were found to be lower. The levels of leukocyte CyP mRNA were increased with exacerbation of clinical disease. Determination of the levels of leukocyte CyP mRNA expression in children with different clinical types showed that there was no difference in the levels of CyP mRNA expression in children with prednisone dependent, frequent relapsing or prednisone-resistant types. The level of leukocyte CyP mRNA expression was found to be higher in acute stage groups than in convalescent stage or control groups. CONCLUSIONS: Determination of the levels of leukocyte CyP mRNA may be of assistance in cyclosporine A therapy. PMID- 16354248 TI - Fatal case of Aspergillus coinfection in a renal transplant recipient suffering from cytomegalovirus pneumonitis. AB - Cytomegalovirus (CMV) disease is common in postrenal transplant recipients, and may predispose the patients to secondary bacterial or fungal infections. However, simultaneous coinfection is rare and often makes diagnosis difficult. We report a case of CMV pneumonitis in a renal transplant recipient presenting with elevated CMV pp65 antigen level and abnormal chest radiograph. Despite potent and broad spectrum antimicrobial therapy, his condition deteriorated rapidly - he soon went into respiratory failure, septic shock and died several days later. Transbronchial biopsy and bronchoalveolar lavage obtained before the patient's death showed evidence of invasive pulmonary aspergillosis with concomitant CMV pneumonitis. High index of suspicion and early and empirical initiation of antifungal therapy may be necessary for successful management of fulminant pneumonia in solid organ transplant recipients. PMID- 16354249 TI - Mega dose of vitamin C augments the nephrotoxicity of paracetamol. PMID- 16354250 TI - Bilateral emphysematous pyelonephritis. PMID- 16354251 TI - Tanning and skin cancer. AB - Skin cancer is a large and growing problem in the United States. Sun and other ultraviolet (UV) light exposures play a key role in the development of skin cancer. Pediatricians can play an important role in counseling patients and are in a position to help educate children and their families about skin cancer. The purpose of this review is to familiarize pediatricians with the magnitude of the skin cancer problem and the evidence that ultraviolet light exposure, particularly indoor tanning, contributes to this problem. We reviewed the literature on ultraviolet light and skin cancer (based on a MEDLINE search of articles using the headings "ultraviolet light" and "skin cancer") and found that skin cancer is the most rapidly growing cause of cancer deaths in the United State. There is strong epidemiologic evidence for the relationship between UV exposure and nonmelanoma skin cancer and growing evidence for the relationship between indoor tanning and melanoma. We recommend that pediatricians counsel children and their parents about UV protection. Measures such as use of sunscreen and hats for outdoor play, both at home and in school, should be encouraged. PMID- 16354252 TI - Sun protection policies in Miami-Dade County public schools: opportunities for skin cancer prevention. AB - Childhood exposure to ultraviolet radiation from the sun and a history of sunburns are risk factors for skin cancer. Because children spend time outdoors when they are at school, school sun protection policies are an important health issue, particularly in areas of the country with year-round warm and sunny climates, such as Florida. To better understand the sun protection policies and practices in South Florida schools, a sample (n = 51) of elementary and middle schools in Miami-Dade County public schools were surveyed as part of a CDC-funded cancer control program at the University of Miami. Of the principals and teachers surveyed, most (78%) knew about the county school system's guidelines for avoiding excessive heat exposure, which include two sun protection measures. Two thirds reported that they shared these guidelines with teachers; 21% shared them with parents. Few schools monitor implementation of the guidelines, although 70% schedule outdoor activities to avoid peak sun hours. No schools required sunscreen, hats, or protective clothing. Physical education teachers and students spend an average of 4.5 and 0.6 hours per day outdoors, respectively. Improved school sun protection policies and monitoring of such policies is needed to reduce sun exposure and skin cancer risk for both students and staff. PMID- 16354253 TI - Children and sun exposure in the northeast of Italy. AB - Excessive, incorrect exposure to the sun at a young age can be a risk factor for skin cancer at a later age. In this study we have investigated the exposure and protection habits of a group of children to see whether there are behavioral errors that should and possibly could be corrected. We handed out a multiple choice questionnaire to 310 subjects, 212 boys and 98 girls between the ages of 6 and 14, all residents in the province of Udine in the northeastern part of Italy. Of these children, 24% had sunburn on several occasions; boys were four times more at risk than girls (OR = 0.4) and the frequency reduced by 30% for each skin phototype class higher. In children between 11 and 14 years of age, exposure was prolonged (43% for 2-4 hours, 38% for 4-8 hours) and 30% preferred peak hours. Eighty percent of the children, mostly girls, younger children (6-10), and the lower skin phototypes used sunscreens, but only 38% used them on a regular basis, whereas 20% applied them after sunbathing and 42% used a single daily application. Two percent of the children used sunglasses and 5% wore a T-shirt regularly. The dermatologist and pediatrician play an important role in advising parents that the sun is good for children, but appropriate measures must be taken to reduce any risks, both immediately and in the long term. PMID- 16354254 TI - Sweet syndrome in two children. AB - We report a 9-month-old girl and a 4-year-old boy with acute febrile neutrophilic dermatosis (Sweet syndrome). Both children were febrile, had leukocytosis, and exhibited lesions characteristic of Sweet syndrome. Both had an antecedent infection. Our evaluation and long-term follow-up of these children failed to reveal evidence of underlying malignancy or a chronic systemic illness typically encountered in Sweet syndrome. Of interest, the 4-year-old boy responded to systemic corticosteroids with remission, whereas the 9-month-old infant experienced flaring of the disease on successive attempts to taper the systemic corticosteroids. Systemic corticosteroid usage was associated with alteration in behavior in the 4-year-old and transient growth retardation in the 9-month-old. In both patients, the adverse effects resolved after discontinuation of the corticosteroids. PMID- 16354255 TI - Pediatric Sweet syndrome and immunodeficiency successfully treated with intravenous immunoglobulin. AB - Painful erythematous plaques and a pathognomonic, dense, dermal infiltrate of neutrophils characterize Sweet syndrome. Although numerous instances of this disease have been reported in children, only four have had an associated immunodeficiency to date. Sweet syndrome has been traditionally treated with systemic corticosteroids despite an unknown pathogenesis. Prior reports suggest that immunodeficient children with Sweet syndrome may have a persistent course and fail to respond to standard treatment. We describe a child with debilitating Sweet syndrome and a primary immunodeficiency whose symptoms were controlled with intravenous immunoglobulin and dapsone. PMID- 16354257 TI - Ichthyosis associated with ARC syndrome: ARC syndrome is one of the differential diagnoses of ichthyosis. AB - The arthrogryposis, renal tubular dysfunction and cholestasis syndrome is rare. Novel identification of the mutation in VPS33B in this syndrome, which involves intracellular protein trafficking by regulation of vesicle-to-target sensory nerve action potential receptor (SNARE) family, might explain the consistent combination of membrane fusion defects. We can guess the defective lamellar body secretion mediated by the soluble N-ethylmaleimide-sensitive factor attachment protein receptor or SNARE protein pathway in the epidermis might result in the ichthyosiform phenotype. In the English-language literature, half of the reported instances of this syndrome are associated with ichthyosis. We report an infant with ichthyosis in association with arthrogryposis, renal tubular dysfunction, and cholestasis syndrome, and review the literature. Our findings suggest that the differential diagnosis of ichthyosis during infancy should include this syndrome. PMID- 16354256 TI - Tinea faciei in an infant caused by Microsporum gypseum simulating a dry impetigo. AB - We report a 14-month-old White boy who was referred to our dermatology unit for evaluation of a skin eruption on his nose. The initial examination led us to the clinical diagnosis of impetigo. The mycologic studies found an uncommon dermatophyte agent, Microsporum gypseum. The main differential diagnosis of tinea faciei is discussed. PMID- 16354258 TI - Atypical pyoderma gangrenosum as a manifestation of childhood acute lymphoblastic leukemia. AB - Pyoderma gangrenosum is a neutrophilic dermatosis that may occur idiopathically or in association with various systemic diseases and malignancy. Although the association of this entity with myeloid malignancies is well known, its association with lymphoid malignancy is extremely rare. We describe atypical pyoderma gangrenosum in association with acute lymphoblastic leukemia in a 2-year old child, an occurrence not reported before. PMID- 16354259 TI - Agminated Spitz nevi: report of a child with a unique dermatomal distribution. AB - Spitz nevi most commonly present as solitary lesions. Multiple agminated Spitz nevi are a rare presentation, with 38 reported cases in the English language literature. We report a 2-year-old girl who presented with multiple Spitz nevi in a unique, dermatome-like distribution and review the English-language literature on agminated Spitz nevi. PMID- 16354260 TI - Xanthoma disseminatum with hepatic involvement in a child. AB - Xanthoma disseminatum is a rare, usually self-healing dermatologic disease of unknown etiology. Involvement of other organs and tissues including bone marrow, bone, and brain may be seen rarely in children. However, to date, hepatic involvement has not been reported. We describe a child with xanthoma disseminatum who had hepatic involvement, and discuss his course and treatment with steroid and azathioprine. PMID- 16354261 TI - An unusual presentation of mastocytosis: unilateral swelling of the vulva. AB - Mastocytosis is a primary, abnormal accumulation of mast cells associated with a broad range of local and systemic symptoms. We report two female adolescents with episodic, unilateral, swelling of the labia majora that was discovered to be an unusual presentation of mastocytosis. Mastocytosis is frequently misdiagnosed because of its rarity and variable clinical presentation, which often mimics other conditions. In the appropriate setting, mastocytosis should be included in the differential diagnosis of labium majus swelling. PMID- 16354262 TI - Persistent B-cell lymphopenia, multiorgan disease, and erythema multiforme caused by Mycoplasma pneumoniae infection. AB - We report a 6-year-old girl in whom Mycoplasma pneumoniae infection presenting with erythema multiforme, multiorgan, and hematologic dysfunctions induced a long standing, marked B-cell lymphopenia. An increase of CD8+ lymphocytes was also detected. We suggest that a selective cytotoxic T lymphocyte-dependent B cell lysis and the expansion of super-antigen activated CD8+ T cells may account for the multiorgan and hematologic disturbances triggered by M. pneumoniae. PMID- 16354263 TI - Successful acne management in Apert syndrome twins. AB - Apert syndrome, or acrocephalosyndactyly, is characterized by craniosynostosis and early epiphyseal closure resulting in various deformities of the skull, hands, and feet. Typically a sporadic condition, autosomal dominant inheritance with complete penetrance has been known to occur. Most adolescents with the disorder are prone to the development of severe pustular facial and truncal acne, with extension to the upper arms and forearm. We report twin brothers with Apert syndrome who, after 2 years of standard management by their pediatrician, were referred for management of complicated acne. In our patients there were a constellation of findings consistent with the disorder and, of importance to this report, significant dermatological manifestations. On presentation, each brother was found to have acne vulgaris of a different stage. Our patients were refractory to conventional treatment for acne but one required and had a significant response to isotretinoin. The risk/benefit ratio in treating acne lesions with isotretinoin in a teenager with Apert syndrome is reviewed. PMID- 16354264 TI - Purpura after application of EMLA cream in two children. AB - The eutectic mixture of local anesthetic cream, a 1 : 1 mixture of prilocaine and lidocaine, 2.5% each, is frequently used in pediatric and dermatologic practice to obtain local anesthesia. Side effects include transient skin blanching, erythema, urticaria, allergic contact dermatitis, irritant contact dermatitis, hyperpigmentation, and purpura. We report two children with a purpuric reaction after application of this mixture cream. Purpura after application of this anesthetic cream is a rare nonallergic reaction and only 17 occurrences have been reported, to our knowledge, in the literature. Patch tests could not be performed in our two patients because of lack of parental consent but we suggest that the purpuric reactions were most probably of toxic origin. The pathogenesis of purpura after application of eutectic mixture of local anesthetics cream, which resolves within 2 weeks without dermatologic sequelae and without any specific therapy, is complex. The lesions are probably caused by the direct effect of the cream components on the vessels but many other factors, such as atopic dermatitis, prematurity, subjective predisposition to purpura, trauma, and thrombocytopenia may play important pathogenetic roles. PMID- 16354265 TI - What syndrome is this? Sjogren-Larsson syndrome. PMID- 16354268 TI - Hyperpigmented patch associated to heterochromia iridis. PMID- 16354270 TI - Wiedemann-Rautenstrauch or trichorhinophalangeal syndrome? PMID- 16354271 TI - Sjogren-Larsson syndrome: Indian and Central European similarities. PMID- 16354272 TI - Pseudoacne of the nasal crease in a patient with congenital deafness and pigmentary mosaicism. PMID- 16354273 TI - Do gender-specific differences in eyelash length in 5- to 6-year-old preschoolchildren with and without atopic eczema exist? Results from the Miriam study conducted in Augsburg, Germany. PMID- 16354276 TI - An overview of regular dialysis treatment in Japan as of 31 December 2003. AB - A statistical survey of 3750 nationwide dialysis facilities was carried out by the Japanese Society for Dialysis Therapy (JSDT) at the end of 2003, with answers to the questionnaires received from 3717 facilities (99.12%). The population of dialysis patients in Japan at the end of 2003 was 237,710, and the number of dialysis patients per million people was 1862.7. The crude death rate during a 1 year period from the end of 2002 to the end of 2003 was 9.3%. The mean age of patients newly introduced to dialysis was 65.4 years, and the mean age of the entire dialysis population was 62.3 years. The primary diseases in the patients newly introduced to dialysis in 2003 included diabetic nephropathy (41.0% of patients) and chronic glomerulonephritis (29.1% of patients). The mean serum neutral fat concentration for all the dialysis patients was 113.9 +/- 71.7 mg/dL (+/- SD). The mean serum low density lipoprotein (LDL)-cholesterol concentration was 90.8 +/- 30.9 mg/dL. Dialysate calcium concentrations ranging from 3.0 mEq/L to less than 3.5 mEq/L were used for majority of the dialysis patients (55.4%). Among anticoagulants given to the dialysis patients, heparins were the most commonly used in 79.3% of the dialysis patients. The relationship between blood pressure during dialysis and life expectancy for 1 year was analyzed for 43,465 patients who had undergone dialysis three times per week at the end of 2001. Results showed a significantly high mortality risk for patients who had systolic blood pressure of less than 100 mm Hg at the start of dialysis, systolic blood pressure of less than 100 mm Hg at the end of dialysis, and the greatest decrease (lowest) in systolic blood pressure of less than 120 mm Hg during dialysis. Patients who received vasopressor therapy during dialysis had a higher mortality risk than those who received no vasopressor therapy. PMID- 16354277 TI - Therapeutic apheresis--state of the art in the year 2005. AB - Therapeutic apheresis is an extracorporeal blood purification method for the treatment of diseases in which pathological proteins or cells have to be eliminated. Selective plasma processing is more efficient in pathogen removal than unselective plasma exchange and does not require a substitution fluid like albumin. This overview presents the various selective devices for the treatment of plasma (plasmapheresis) and blood cells (leukocyte apheresis). Prospective randomized trials were performed for the treatment of age-related macular degeneration (Rheopheresis), sudden hearing loss (heparin-induced lipoprotein precipitation [HELP]), rheumatoid arthritis (Prosorba), dilative cardiomyopathy (Ig-Therasorb, Immunosorba), acute-on-chronic liver failure (molecular adsorbent recirculating system [MARS]), and ulcerative colitis (Cellsorba). Prospective non randomized controlled trials were carried out treating hypercholesterolemia (Liposorber) and crossmatch-positive recipients before kidney transplantation (Immunosorba). Uncontrolled studies were done for ABO-incompatibility in living donor kidney transplantation (KT) (Glycosorb), acute humoral rejection after KT (Immunosorba) and acute liver failure (Prometheus). According to the 2002 International Apheresis Registry covering 11428 sessions in 811 patients, 79% of the patients showed an improvement of their condition by apheresis and only a few sessions were fraught with adverse effects (AE). The major AE were blood access difficulties (3.1%) and hypotension (1.6%). In summary, therapeutic apheresis is a safe and effective procedure for the treatment of diseases refractory to drug therapy. PMID- 16354278 TI - Direct adsorption of lipoproteins from whole blood by direct adsorption of lipoprotein apheresis: first experience in two hypercholesterolemic children. AB - Low density lipoprotein (LDL) cholesterol apheresis, combined with lipid lowering drugs, provides a safe and effective means of improving the prognosis of patients with homozygote familial hypercholesterolemia, especially if started before the age of seven. The direct adsorption of lipoprotein (DALI) is the first extracorporeal low density lipoprotein removing system compatible with whole blood. The purpose of the present study was to clarify the efficacy and safety of DALI in children with homozygous familial hypercholesterolemia. Two boys, aged 9 and 15 years, with familial hypercholesterolemia, who were highly resistant to dietary regimes and to drug therapy, were treated with the low density lipoprotein adsorber DALI apheresis once every 2 weeks for 24 weeks. The treated blood volumes for each procedure were 2911 mL (493 +/- SD) and 5982 mL (1129 +/- SD), respectively. In our patients, the acute mean LDL cholesterol reductions were 44.7 +/- 8.9% and 58.8 +/- 4.5%. The corresponding reductions were 42.5 +/- 7.2% and 56 +/- 4.3% for total cholesterol, and 46.5 +/- 17.1% and 55 +/- 7.5% for very low density lipoprotein cholesterol (VLDL-C). There were insignificant losses of high density lipoprotein (12.2 +/- 5.7%, 8.3 +/- 5.5%). Treatment was well tolerated in general, and neither patient suffered from irreversible or long lasting adverse effects. Our experience with DALI apheresis is encouraging. The present report is the first on the use of DALI in children. Based on this short term evaluation we think that DALI might be safe and effective in children with homozygote familial hypercholesterolemia, however further evaluation of long-term effects is needed. PMID- 16354279 TI - Rheopheresis in patients with critical limb ischemia--results of an open label prospective pilot trial. AB - Rheopheresis is a specifically designed application of double filtration plasmapheresis, for extracorporeal treatment of microcirculatory disorders. Safety and efficacy of Rheopheresis for wound healing and skin oxygenation were investigated in patients with critical limb ischemia. Twelve patients of Fontaine stage III-IV were treated with a series of 10 Rheopheresis sessions over 17 weeks. Transcutaneous oxygen pressure (tcpO(2)) and ankle-brachial index (ABI) were repeatedly determined to monitor the effects of the Rheopheresis treatment series on microcirculation and skin blood flow. Laboratory parameters of blood rheology were measured in addition to safety parameters and course of the pain syndrome was documented. In four patients (baseline Fontaine stage III) Rheopheresis was associated with an improvement of Fontaine stage, a pronounced increase in tcpO(2) and complete regression of the rest pain. As an adjunct therapeutic option, Rheopheresis may preserve a functional lower extremity, delay amputation or reduce the extent of amputation. PMID- 16354280 TI - Abstracts of the 28th Study Group on IgA Nephropathy, Tokyo, Japan, 29 January 2005. PMID- 16354294 TI - Geostatistical analysis of disease data: estimation of cancer mortality risk from empirical frequencies using Poisson kriging. AB - BACKGROUND: Cancer mortality maps are used by public health officials to identify areas of excess and to guide surveillance and control activities. Quality of decision-making thus relies on an accurate quantification of risks from observed rates which can be very unreliable when computed from sparsely populated geographical units or recorded for minority populations. This paper presents a geostatistical methodology that accounts for spatially varying population sizes and spatial patterns in the processing of cancer mortality data. Simulation studies are conducted to compare the performances of Poisson kriging to a few simple smoothers (i.e. population-weighted estimators and empirical Bayes smoothers) under different scenarios for the disease frequency, the population size, and the spatial pattern of risk. A public-domain executable with example datasets is provided. RESULTS: The analysis of age-adjusted mortality rates for breast and cervix cancers illustrated some key features of commonly used smoothing techniques. Because of the small weight assigned to the rate observed over the entity being smoothed (kernel weight), the population-weighted average leads to risk maps that show little variability. Other techniques assign larger and similar kernel weights but they use a different piece of auxiliary information in the prediction: global or local means for global or local empirical Bayes smoothers, and spatial combination of surrounding rates for the geostatistical estimator. Simulation studies indicated that Poisson kriging outperforms other approaches for most scenarios, with a clear benefit when the risk values are spatially correlated. Global empirical Bayes smoothers provide more accurate predictions under the least frequent scenario of spatially random risk. CONCLUSION: The approach presented in this paper enables researchers to incorporate the pattern of spatial dependence of mortality rates into the mapping of risk values and the quantification of the associated uncertainty, while being easier to implement than a full Bayesian model. The availability of a public domain executable makes the geostatistical analysis of health data, and its comparison to traditional smoothers, more accessible to common users. In future papers this methodology will be generalized to the simulation of the spatial distribution of risk values and the propagation of the uncertainty attached to predicted risks in local cluster analysis. PMID- 16354295 TI - Detection of cold pain, cold allodynia and cold hyperalgesia in freely behaving rats. AB - BACKGROUND: Pain is elicited by cold, and a major feature of many neuropathic pain states is that normally innocuous cool stimuli begin to produce pain (cold allodynia). To expand our understanding of cold induced pain states we have studied cold pain behaviors over a range of temperatures in several animal models of chronic pain. RESULTS: We demonstrate that a Peltier-cooled cold plate with +/ 1 degrees C sensitivity enables quantitative measurement of a detection withdrawal response to cold stimuli in unrestrained rats. In naive rats the threshold for eliciting cold pain behavior is 5 degrees C. The withdrawal threshold for cold allodynia is 15 degrees C in both the spared nerve injury and spinal nerve ligation models of neuropathic pain. Cold hyperalgesia is present in the spared nerve injury model animals, manifesting as a reduced latency of withdrawal response threshold at temperatures that elicit cold pain in naive rats. We also show that following the peripheral inflammation produced by intraplantar injection of complete Freund's adjuvant, a hypersensitivity to cold occurs. CONCLUSION: The peltier-cooled provides an effective means of assaying cold sensitivity in unrestrained rats. Behavioral testing of cold allodynia, hyperalgesia and pain will greatly facilitate the study of the neurobiological mechanisms involved in cold/cool sensations and enable measurement of the efficacy of pharmacological treatments to reduce these symptoms. PMID- 16354296 TI - Implementation failures in the use of two New Zealand laws to control the tobacco industry: 1989-2005. AB - BACKGROUND: We reviewed the implementation of New Zealand laws in relation to the activities of the tobacco industry and their allies. Material for two brief case studies was obtained from correspondence with official agencies, official information requests, internet searches (tobacco industry documents and official government sites), and interviews with 12 key informants. RESULTS: The first case study identified four occasions over a period of 14 years where New Zealand Government agencies appeared to fail to enforce consumer protection law, although apparent breaches by the tobacco industry and their allies had occurred in relation to statements on the relative safety of secondhand smoke. The second case study examined responses to a legal requirement for the tobacco industry to provide information on tobacco additives. There was failure to enforce the law, and a failure of the political process for at least 13 years to clarify and strengthen the law. Relevant factors in both these cases of 'policy slippage' appear to have been financial and opportunity costs of taking legal action, political difficulties and the fragmented nature of government structures. CONCLUSION: Considered together, these case studies suggest the need for governments to: (i) make better use of national consumer laws (with proper monitoring and enforcement) in relation to tobacco; and (ii) to strengthen international law and resources around tobacco-related consumer protection. A number of options for achieving these aims are available to governments. PMID- 16354297 TI - Probabilistic annotation of protein sequences based on functional classifications. AB - BACKGROUND: One of the most evident achievements of bioinformatics is the development of methods that transfer biological knowledge from characterised proteins to uncharacterised sequences. This mode of protein function assignment is mostly based on the detection of sequence similarity and the premise that functional properties are conserved during evolution. Most automatic approaches developed to date rely on the identification of clusters of homologous proteins and the mapping of new proteins onto these clusters, which are expected to share functional characteristics. RESULTS: Here, we inverse the logic of this process, by considering the mapping of sequences directly to a functional classification instead of mapping functions to a sequence clustering. In this mode, the starting point is a database of labelled proteins according to a functional classification scheme, and the subsequent use of sequence similarity allows defining the membership of new proteins to these functional classes. In this framework, we define the Correspondence Indicators as measures of relationship between sequence and function and further formulate two Bayesian approaches to estimate the probability for a sequence of unknown function to belong to a functional class. This approach allows the parametrisation of different sequence search strategies and provides a direct measure of annotation error rates. We validate this approach with a database of enzymes labelled by their corresponding four-digit EC numbers and analyse specific cases. CONCLUSION: The performance of this method is significantly higher than the simple strategy consisting in transferring the annotation from the highest scoring BLAST match and is expected to find applications in automated functional annotation pipelines. PMID- 16354298 TI - A decoy set for the thermostable subdomain from chicken villin headpiece, comparison of different free energy estimators. AB - BACKGROUND: Estimators of free energies are routinely used to judge the quality of protein structural models. As these estimators still present inaccuracies, they are frequently evaluated by discriminating native or native-like conformations from large ensembles of so-called decoy structures. RESULTS: A decoy set is obtained from snapshots taken from 5 long (100 ns) molecular dynamics (MD) simulations of the thermostable subdomain from chicken villin headpiece. An evaluation of the energy of the decoys is given using: i) a residue based contact potential supplemented by a term for the quality of dihedral angles; ii) a recently introduced combination of four statistical scoring functions for model quality estimation (FRST); iii) molecular mechanics with solvation energy estimated either according to the generalized Born surface area (GBSA) or iv) the Poisson-Boltzmann surface area (PBSA) method. CONCLUSION: The decoy set presented here has the following features which make it attractive for testing energy scoring functions:1) it covers a broad range of RMSD values (from less than 2.0 A to more than 12 A);2) it has been obtained from molecular dynamics trajectories, starting from different non-native-like conformations which have diverse behaviour, with secondary structure elements correctly or incorrectly formed, and in one case folding to a native-like structure. This allows not only for scoring of static structures, but also for studying, using free energy estimators, the kinetics of folding;3) all structures have been obtained from accurate MD simulations in explicit solvent and after molecular mechanics (MM) energy minimization using an implicit solvent method. The quality of the covalent structure therefore does not suffer from steric or covalent problems. The statistical and physical effective energy functions tested on the set behave differently when native simulation snapshots are included or not in the set and when averaging over the trajectory is performed. PMID- 16354299 TI - Technical efficiency of peripheral health units in Pujehun district of Sierra Leone: a DEA application. AB - BACKGROUND: The Data Envelopment Analysis (DEA) method has been fruitfully used in many countries in Asia, Europe and North America to shed light on the efficiency of health facilities and programmes. There is, however, a dearth of such studies in countries in sub-Saharan Africa. Since hospitals and health centres are important instruments in the efforts to scale up pro-poor cost effective interventions aimed at achieving the United Nations Millennium Development Goals, decision-makers need to ensure that these health facilities provide efficient services. The objective of this study was to measure the technical efficiency (TE) and scale efficiency (SE) of a sample of public peripheral health units (PHUs) in Sierra Leone. METHODS: This study applied the Data Envelopment Analysis approach to investigate the TE and SE among a sample of 37 PHUs in Sierra Leone. RESULTS: Twenty-two (59%) of the 37 health units analysed were found to be technically inefficient, with an average score of 63% (standard deviation = 18%). On the other hand, 24 (65%) health units were found to be scale inefficient, with an average scale efficiency score of 72% (standard deviation = 17%). CONCLUSION: It is concluded that with the existing high levels of pure technical and scale inefficiency, scaling up of interventions to achieve both global and regional targets such as the MDG and Abuja health targets becomes far-fetched. In a country with per capita expenditure on health of about USD 7, and with only 30% of its population having access to health services, it is demonstrated that efficiency savings can significantly augment the government's initiatives to cater for the unmet health care needs of the population. Therefore, we strongly recommend that Sierra Leone and all other countries in the Region should institutionalize health facility efficiency monitoring at the Ministry of Health headquarter (MoH/HQ) and at each health district headquarter. PMID- 16354300 TI - Encapsulated Escherichia coli in alginate beads capable of secreting a heterologous pectin lyase. AB - BACKGROUND: Production of heterologous proteins in the E. coli periplasm, or into the extracellular fluid has many advantages; therefore naturally occurring signal peptides are selected for proteins translocation. The aim of this study was the production in high yields of a recombinant pectin lyase that is efficiently secreted and the encapsulation of transformed E. coli cells for pectin degradation in a biotechnological process. RESULTS: The nucleotide sequence of Bacillus subtilis alpha-amylase's signal peptide was fused to the N-terminal of an heterologously expressed pectin lyase in E. coli BL21 [DE3]. Thus pectin lyase secretion was achieved into the extracellular growth medium. E. coli cells harboring the recombinant plasmid heterologously express pectin lyase to around 22% of the total cellular proteins, as it was estimated by SDS-PAGE and image analysis. IPTG induces the heterologously expressed enzyme, which is initially distributed extracellularly (7 hour) and later on at the periplasmic (9 hours) or cytosolic fraction (20 hours). No pectin lyase activity was found in the membranes fraction and in the inclusion bodies. Encapsulation of the recombinant strains of E. coli in alginate or alginate/silica beads 1:5 showed that pectin lyase could degrade effectively its substrate, for at least ten operational cycles. CONCLUSION: Secretion of an heterologously overexpressed pectin lyase in E. coli BL21 [DE3] was achieved in this study. For this purpose the signal peptide of alpha-amylase from B. subtilis was fused to the N-terminal domain of pectin lyase. Encapsulated E. coli BL21 [DE3] cells harboring pET29c/exPNL were used successfully for pectin degradation up to ten operational cycles indicating that under special conditions this might have biotechnological implementations. PMID- 16354301 TI - Experimental glomerulonephritis induced by hydrocarbon exposure: a systematic review. AB - BACKGROUND: Much epidemiological evidence suggests that hydrocarbon exposure may induce glomerulonephritis and worsen its course in many patients. The mechanisms are unknown, however, no specific microscopic pattern has been identified, and it has also been argued that hydrocarbon exposure causes tubular damage mainly. Studying experimental animals may best answer these questions, and as no systematic review of glomerulonephritis produced experimentally by hydrocarbon exposure has been performed previously, I found it relevant to search for and analyse such studies. METHODS: Animal experiments having mimicked human glomerulonephritis by hydrocarbon exposure were sought on Medline and Toxnet RESULTS: Twenty-six experiments using thirteen different hydrocarbons were identified. Several human subtypes were observed including IgA nephritis, mesangial, proliferative and extracapillary glomerulonephritis, focal and focal segmental sclerosis, minimal change nephropathy, anti-GBM and anti-TBM nephritis, and glomerulonephritis associated with peiarteritis nodosa. Glomerular proteinuria was seen in 10/12 experiments that included urine analyses, and renal failure in 5/8 experiments that included measurements of glomerular function. All experiments resulted in various degrees of tubular damage as well. In most studies, where the animals were examined at different times during or after the exposure, the renal microscopic and functional changes were seen immediately, whereas deposits of complement and immunoglobulins appeared late in the course, if at all. CONCLUSION: These experiments are in accord with epidemiological evidence that hydrocarbon exposure may cause glomerulonephritis and worsen renal function. Probable mechanisms include an induction of autologous antibodies and a disturbance of normal immunological functions. Also, tubular damage may increase postglomerular resistance, resulting in a glomerular deposition of macromolecules. In most models a causal role of glomerular immune complex formation was unlikely, but may rather have been a secondary phenomenon. As most glomerulonephritis subgroups were seen and as some of the hydrocarbons produced more than one subgroup, the microscopic findings in a patient cannot be used as a clue to the causation of his disease. By the same reason, the lack of a specific histological pattern in patients with glomerulonephritis assumed to have been caused by hydrocarbon exposure is not contradictive. PMID- 16354302 TI - Exclusion of PINK1 as candidate gene for the late-onset form of Parkinson's disease in two European populations. AB - BACKGROUND: Parkinson's disease (PD) is the second most common neurodegenerative disorder. Recently, mutations in the PINK1 (PARK6) gene were shown to rarely cause autosomal-recessively transmitted, early-onset parkinsonism. In order to evaluate whether PINK1 contributes to the risk of common late-onset PD we analysed PINK1 sequence variations. A German (85 patients) and a Norwegian cohort (90 patients) suffering from late-onset PD were screened for mutations and single nucleotide polymorphisms (SNPs) in the PINK1 gene. Both cohorts consist of well characterized patients presenting a positive family history of PD in approximately 17%. Investigations were performed by single strand conformation polymorphism (SSCP), denaturating high performance liquid chromatography (DHPLC) and sequencing analyses. SNP frequencies were compared by the chi2 test. RESULTS: Several common SNPs were identified in our cohorts, including a recently identified coding variant (Q115L) in exon 1. Genotyping of the Q115L variation did not reveal significant frequency differences between patients and controls. Pathogenic mutations in the PINK1 gene were not identified, neither in the German nor in the Norwegian cohort. CONCLUSION: Sequence variation in the PINK1 gene appears to play a marginal quantitative role in the pathogenesis of the late onset form of PD, in German and Norwegian cohorts, if at all. PMID- 16354303 TI - Indirect comparison of interventions using published randomised trials: systematic review of PDE-5 inhibitors for erectile dysfunction. AB - BACKGROUND: There are no randomised and properly blinded trials directly comparing one PDE-5 inhibitor with another in a normal home setting. Valid indirect comparisons with a common comparator must examine equivalent doses, similar duration, similar populations, with the same outcomes reported in the same way. METHODS: Published randomised, double-blind trials of oral PDE-5 inhibitors for erectile dysfunction were sought from reference lists in previous reviews and electronic searching. Analyses of efficacy and harm were carried out for each treatment, and results compared where there was a common comparator and consistency of outcome reporting, using equivalent doses. RESULTS: Analysis was limited by differential reporting of outcomes. Sildenafil trials were clinically and geographically more diverse. Tadalafil and vardenafil trials tended to use enriched enrollment. Using all trials, the three interventions were similar for consistently reported efficacy outcomes. Rates of successful intercourse for sildenafil, tadalafil and vardenafil were 65%, 62%, and 59%, with placebo rates of 23-28%. The rates of improved erections were 76%, 75% and 71%, respectively, with placebo rates of 22-24%, and NNTs of 1.9 or 2.0. Reporting of withdrawals was less consistent, but all-cause withdrawals for sildenafil, tadalafil and vardenafil were 8% 13% and 20%. All three drugs were well tolerated, with headache being the most commonly reported event at 13-17%. There were few serious adverse events. CONCLUSION: There were differences between trials in outcomes reported, limiting comparisons, and the most useful outcomes were not reported. For common outcomes there was similar efficacy between PDE-5 inhibitors. PMID- 16354304 TI - X-ray structure of engineered human Aortic Preferentially Expressed Protein-1 (APEG-1). AB - BACKGROUND: Human Aortic Preferentially Expressed Protein-1 (APEG-1) is a novel specific smooth muscle differentiation marker thought to play a role in the growth and differentiation of arterial smooth muscle cells (SMCs). RESULTS: Good quality crystals that were suitable for X-ray crystallographic studies were obtained following the truncation of the 14 N-terminal amino acids of APEG-1, a region predicted to be disordered. The truncated protein (termed DeltaAPEG-1) consists of a single immunoglobulin (Ig) like domain which includes an Arg-Gly Asp (RGD) adhesion recognition motif. The RGD motif is crucial for the interaction of extracellular proteins and plays a role in cell adhesion. The X ray structure of DeltaAPEG-1 was determined and was refined to sub-atomic resolution (0.96 A). This is the best resolution for an immunoglobulin domain structure so far. The structure adopts a Greek-key beta-sandwich fold and belongs to the I (intermediate) set of the immunoglobulin superfamily. The residues lying between the beta-sheets form a hydrophobic core. The RGD motif folds into a 310 helix that is involved in the formation of a homodimer in the crystal which is mainly stabilized by salt bridges. Analytical ultracentrifugation studies revealed a moderate dissociation constant of 20 microM at physiological ionic strength, suggesting that APEG-1 dimerisation is only transient in the cell. The binding constant is strongly dependent on ionic strength. CONCLUSION: Our data suggests that the RGD motif might play a role not only in the adhesion of extracellular proteins but also in intracellular protein-protein interactions. However, it remains to be established whether the rather weak dimerisation of APEG-1 involving this motif is physiologically relevant. PMID- 16354305 TI - Policy lessons from comparing mortality from two global forces: international terrorism and tobacco. AB - BACKGROUND: The aim of this study was to compare the mortality burdens from two global impacts on mortality: international terrorism and the major cause of preventable death in developed countries--tobacco use. We also sought to examine the similarities and differences between these two causes of mortality so as to better inform the policy responses directed at prevention. METHODS: Data on deaths from international terrorism were obtained from a US State Department database for 1994-2003. Estimates for tobacco-attributable deaths were based on Peto et al 2003. The countries were 37 developed and East European countries. RESULTS AND DISCUSSION: The collective annualized mortality burden from tobacco was approximately 5700 times that of international terrorism. The ratio of annual tobacco to international terrorism deaths was lowest for the United States at 1700 times, followed by Russia at 12,900 times. The tobacco death burden in all these countries was equivalent to the impact of an 11 September type terrorist attack every 14 hours. Different perceptions of risk may contribute to the relative lack of a policy response to tobacco mortality, despite its relatively greater scale. The lack is also despite tobacco control having a stronger evidence base for the prevention measures used. CONCLUSION: This comparison highlights the way risk perception may determine different policy responses to global forces causing mortality. Nevertheless, the large mortality differential between international terrorism and tobacco use has policy implications for informing the rational use of resources to prevent premature death. PMID- 16354306 TI - A balanced transcription between telomerase and the telomeric DNA-binding proteins TRF1, TRF2 and Pot1 in resting, activated, HTLV-1-transformed and Tax expressing human T lymphocytes. AB - BACKGROUND: The functional state of human telomeres is controlled by telomerase and by a protein complex named shelterin, including the telomeric DNA-binding proteins TRF1, TRF2 and Pot1 involved in telomere capping functions. The expression of hTERT, encoding the catalytic subunit of telomerase, plays a crucial role in the control of lymphocyte proliferation by maintaining telomere homeostasis. It has been previously found that hTERT activity is down-regulated by the human T cell leukaemia virus type 1 (HTLV-1) Tax protein in HTLV-1 transformed T lymphocytes. In this study, we have examined the effects of Tax expression on the transcriptional profile of telomerase and of shelterin in human T lymphocytes. RESULTS: We first provide evidence that the up-regulation of hTERT transcription in activated CD4+ T lymphocytes is associated with a down regulation of that of TERF1, TERF2 and POT1 genes. Next, the down-regulation of hTERT transcription by Tax in HTLV-1 transformed or in Tax-expressing T lymphocytes is found to correlate with a significant increase of TRF2 and/or Pot1 mRNAs. Finally, ectopic expression of hTERT in one HTLV-1 T cell line induces a marked decrease in the transcription of the POT1 gene. Collectively, these observations predict that the increased transcriptional expression of shelterin genes is minimizing the impact on telomere instability induced by the down regulation of hTERT by Tax. CONCLUSION: These findings support the notion that Tax, telomerase and shelterin play a critical role in the proliferation of HTLV-1 transformed T lymphocytes. PMID- 16354308 TI - Treatment options for patients with advanced-stage non-small-cell lung cancer: who are the "elderly" and who are the "unfit"? PMID- 16354307 TI - Use of different but overlapping determinants in a retrovirus receptor accounts for non-reciprocal interference between xenotropic and polytropic murine leukemia viruses. AB - BACKGROUND: Retrovirus infection depends on binding of the retroviral envelope (Env) protein to specific cell-surface protein receptors. Interference, or superinfection resistance, is a frequent consequence of retroviral infection, and occurs when newly-synthesized Env binds to receptor proteins resulting in a block to entry by retroviruses that use the same receptors. Three groups of viruses demonstrate a non-reciprocal pattern of interference (NRI), which requires the existence of both a common receptor utilized by all viruses within the group, and a specific receptor that is used by a subset of viruses. In the case of amphotropic and 10A1 murine leukemia viruses (MLV), the common and specific receptors are the products of two related genes. In the case of avian sarcoma and leukosis virus types B, D, and E, the two receptors are distinct protein products of a single gene. NRI also occurs between xenotropic and polytropic MLV. The common receptor, Xpr1, has been identified, but a specific receptor has yet to be described. RESULTS: Using chimeric receptor proteins and interference studies, we have identified a region of Xpr1 that is uniquely utilized by xenotropic MLV and show that this receptor domain is required for non-reciprocal interference. CONCLUSION: We propose a novel pattern of receptor usage by xenotropic and polytropic MLV to explain the NRI observed between these viruses. We propose that the specific and common receptor determinants for xenotropic and polytropic viruses are simultaneously present in discreet domains of a single Xpr1 protein. PMID- 16354309 TI - Relevance of epidermal growth factor receptor genomic gain in predicting clinical response to kinase inhibitors in non-small-cell lung cancer. PMID- 16354310 TI - Undertreatment of elderly patients with non-small-cell lung cancer. AB - The absolute number of patients with lung cancer is rising as a result of our aging population. Until recently, clinicians have been reluctant to aggressively treat elderly patients with non-small-cell lung cancer (NSCLC) because of a lack of supportive data and concern for potential toxicity. Recently, evidence has emerged that suggests that, similar to younger patients, healthy elderly patients can benefit from therapy in all stages of NSCLC. This review will discuss the findings that indicate that chronologic age alone should not be a barrier to appropriate treatment for NSCLC, but consideration should be given to more important prognostic factors such as comorbidities and performance status. PMID- 16354311 TI - Vascular-targeting agents and radiation therapy in lung cancer: where do we stand in 2005? AB - With recent Food and Drug Administration approval of the anti-vascular endothelial growth factor (VEGF) antibody for the treatment of colon cancer, it may be possible to achieve similar progress in the treatment of locally advanced lung cancer. Antiangiogenic therapies in the clinic are a reality, and it is important to demonstrate that they can be used safely with conventional modalities, including radiation therapy (RT). Strategies under scrutiny in preclinical and clinical studies include the use of endogenous inhibitors of angiogenesis, use of agents that target VEGF and VEGF receptor signaling, targeting endothelial-related integrins during angiogenesis, and targeting the preexisting immature vessels growing within tumors (ie, vascular targeting). Regardless of the approach, it is necessary to address whether angiogenesis is a consistent phenomenon within the lung parenchyma around a cancer and a relevant target and whether inhibiting angiogenesis will improve current lung cancer therapies without increasing toxicity. Vascular-targeting agents (VTAs) are an interesting class of agents that have the potential to enhance RT, but their clinical promise has yet to be realized. In preclinical models, these agents selectively destroy the tumor vasculature, initiating a rapid centralized necrosis within established tumors. Characteristically, after treatment with VTAs, a rim of viable tumor cells remains at the periphery of the tumor, which remains well perfused and should therefore be relatively sensitive to radiation induced cytotoxicity. This review will focus on VTAs in the treatment of lung cancer and includes a discussion of combination studies with RT in the laboratory and some of the hurdles in the clinical application of these agents. PMID- 16354312 TI - Influence of sex on lung cancer histology, stage, and survival in a midwestern United States tumor registry. AB - BACKGROUND: A study was performed to identify differences between men and women with regard to lung cancer type, stage at diagnosis, and survival in a single hospital system cancer registry. PATIENTS AND METHODS: A retrospective cohort study was designed based on a study population drawn from the lung cancer tumor registry at a single hospital system composed of 2 independent hospitals in the Midwestern United States. This database included all patients from 1996 to 2002 with known lung cancer or abnormal findings on chest radiography or computed tomography (N=2618). Patients with adenocarcinoma or squamous cell, small-cell, or large-cell carcinoma were included in the study. Data were collected on patient sex, age, cancer type, stage at diagnosis, and survival status. RESULTS: A total of 1216 men and 997 women met inclusion criteria for the study. There was no significant difference in age between sexes at diagnosis. Women were significantly more likely to have adenocarcinoma or small-cell carcinoma but less likely to have squamous cell carcinoma compared with men. There were no significant differences between sexes in the incidence of large-cell carcinoma. No significant differences were found between men and women in terms of cancer stage at diagnosis. There were significant differences in survival between the histologic types at years 3, 4, and 5. Only patients with stage I disease showed a difference between sexes and only for years 2, 3, 4, and 5. CONCLUSION: Overall differences in lung cancer histology and survival were found between men and women. Because a high mortality rate of lung cancer exists in both sexes, it is important to understand its occurrence and survival rates in both sexes. PMID- 16354313 TI - Sequential versus alternating administration of cisplatin/etoposide and topotecan as first-line treatment in extensive-stage small-cell lung cancer: preliminary results of a Phase III Trial of the Hellenic Oncology Research Group. AB - BACKGROUND: This trial was designed to compare the efficacy and toxicity of sequential versus alternating administration of cisplatin/etoposide and topotecan in patients with previously untreated extensive-stage small-cell lung cancer (SCLC). PATIENTS AND METHODS: Two hundred eighty-four chemotherapy-naive patients were randomized between the sequential therapy arm (n=142; 4 cycles of cisplatin 75 mg/m2 intravenously [I.V.] on day 1 with etoposide 100 mg/m2 per day I.V. on days 1-3 followed by 4 cycles of topotecan 1.5 mg/m2 per day I.V. on days 1-5) and the alternating arm (n=142; same doses of cisplatin/etoposide on cycles 1, 3, 5, and 7 and topotecan on cycles 2, 4, 6, and 8). Treatment cycles for both regimens were administered every 3 weeks. RESULTS: At this preliminary analysis, no statistically significant difference in the overall response rate, duration of response, time to disease progression, or median survival was observed between the 2 arms. A total of 756 cycles of the sequential therapy and 830 cycles of the alternating therapy were administered, with a median numbers of 6 and 7 cycles per patient, respectively. Topotecan was administered in 85 patients on the sequential arm and 132 patients on the alternating arm. Dose reductions for toxicity were similar in both arms. Grade 3/4 toxicities in the sequential and alternating arms, respectively, included neutropenia (51% and 52%; P=NS), anemia (12% and 11%; P=NS), febrile neutropenia (7% and 9%; P=NS), thrombocytopenia (19% and 20%; P=NS), and asthenia (8% and 2%; P=0.02). There were 4 toxicity-related deaths in the sequential arm versus 3 in the alternating arm. CONCLUSION: Our preliminary conclusion is that the sequential and alternating regimens resulted in comparable activity and tolerability in previously untreated patients with extensive-stage SCLC. PMID- 16354314 TI - Topotecan in the treatment of elderly patients with relapsed small-cell lung cancer. AB - BACKGROUND: Almost 70% of all patients with lung cancer in the United States are>65 years of age, and the incidence of small-cell lung cancer (SCLC) increases with age until the eighth decade of life. However, elderly patients are underrepresented in clinical trials and are often suboptimally treated. The validity of age as a prognostic factor for toxicity or survival remains controversial. PATIENTS AND METHODS: To investigate the safety and efficacy of topotecan (an approved treatment for relapsed SCLC) in older patients, we performed a retrospective analysis in patients >or= 65 years of age versus patients < 65 years of age from 5 large topotecan trials. In all 5 trials, patients received topotecan 1.5 mg/m2 per day via a 30-minute intravenous infusion on days 1 through 5 of a 21-day cycle. Efficacy and tolerability outcomes were assessed for both age groups. RESULTS: Topotecan was similarly tolerated in both age groups, with generally manageable hematologic toxicity. The incidence, duration, and onset of severe hematologic toxicities did not vary significantly with age. In the<65 age group, grade 4 neutropenia and leukopenia were reported in 72% and 32% of patients, respectively; in the >or= 65 age group, grade 4 neutropenia and leukopenia were reported in 77% and 31% of patients, respectively. Grade 4 thrombocytopenia was less common in the<65 age group. Nonhematologic toxicities, median time to progression, and overall survival were comparable between groups. CONCLUSION: This is the first demonstration of the safety and efficacy of topotecan in older patients with recurrent SCLC. Future studies are needed to fully characterize the role of topotecan in the treatment of older patients. PMID- 16354315 TI - Patterns of recurrence and long-term survival after curative resection of localized fibrous tumors of the pleura. AB - BACKGROUND: Localized fibrous tumors of the pleura (LFTPs) are uncommon thoracic neoplasms with variable malignant potential that were previously classified as benign presentation of mesothelioma. We investigate the clinical presentation, recurrence patterns, and long-term survival of patients undergoing curative resection of LFTPs. PATIENTS AND METHODS: Thirty-four patients underwent resection of LFTPs in the present study. There were 20 women and 14 men with a median age of 59 years (range, 35-81 years). The malignant potential of the tumor was estimated through histologic assessment of the degree of cellularity, mitotic activity, and nuclear pleomorphism. Patients' clinical outcomes were correlated to pathologic findings. RESULTS: Seventeen patients (50%) were symptomatic, and 7 tumors (21%) were considered malignant by histologic characteristics. Ipsilateral pleural recurrence remote to the original tumor site occurred in 6 of these patients with malignant microscopic characteristics at a mean of 9 months after resection. Repeated resection was accomplished in 3 of these patients (range, 3-8 repeat operations). There have been no recurrences among the other 27 patients with benign histologic features, and 31 patients remain alive at a median follow up of 34.5 months. CONCLUSION: Histologic characteristics are helpful in estimating the risk of recurrence among patients with LFTPs. Close surveillance is recommended for patients with malignant histologic characteristics. Local recurrence is common without evidence of systemic spread among such patients. Surgical resection is definitive therapy; however, patients with tumors with histologic atypia require close postoperative observation because of the frequency of locoregional recurrence. PMID- 16354316 TI - Gemcitabine/carboplatin in patients with metastatic non-small-cell lung cancer: phase II study of 28-day and 21-day schedules. AB - BACKGROUND: The combination of gemcitabine/carboplatin is active and is widely used for advanced non-small-cell lung cancer (NSCLC). The optimum schedule for the administration of these agents is not currently known. This study compared 2 different combinations of carboplatin/gemcitabine to identify a schedule with a superior therapeutic index. PATIENTS AND METHODS: The 28-day schedule used gemcitabine 1,100 mg/m2 on days 1 and 8 plus carboplatin to an area under the curve (AUC) of 5 on day 8. The 21-day schedule used gemcitabine 1,000 mg/m2 on days 1 and 8 plus carboplatin to an AUC of 5 on day 1. RESULTS: Four hundred ninety-nine chemotherapy-naive patients with stage IIIB NSCLC (with malignant pleural effusion) or stage IV NSCLC were accrued. Grade 3/4 toxicities with the 28-day and 21-day schedules were anemia (4.8% and 6.9% of cycles, respectively), neutropenia (9.1% and 10.7% of cycles, respectively), and thrombocytopenia (8% and 14.4% of cycles, respectively). Thirty-four patients (13.2%) received blood transfusions with the 28-day regimen, as did 43 (20%) with the 21-day regimen, and no bleeding episodes occurred. Overall response rates were 22.8% with the 28 day schedule and 32.6% with the 21-day schedule (P=0.0338). Median survival and 1 year and 2-year survival rates were 9.8 months, 38.6%, and 15.7%, respectively, with the 28-day schedule and 12 months, 49.1%, and 15.8%, respectively, with the 21-day schedule (P=0.5098). CONCLUSION: Gemcitabine/carboplatin demonstrated efficacy comparable to other platinum agent doublets. Either schedule can be recommended, as differences in survival between groups were not significant. PMID- 16354317 TI - Randomized multicenter phase II study of gemcitabine versus docetaxel as first line therapy with second-line crossover in advanced-stage non-small-cell lung cancer. AB - BACKGROUND: A randomized phase II study was performed to determine whether single agent gemcitabine or docetaxel with the introduction of the opposite agent in case of disease progression (ie, in the second-line setting) is feasible and effective in chemotherapy-naive patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: The doses were 1,000 mg/m2 for gemcitabine and 35 mg/m2 for docetaxel, each given on days 1, 8, and 15 every 4 weeks. After a planned interim analysis, the docetaxel/gemcitabine arm (ie, docetaxel followed by gemcitabine) was closed after enrollment of 49 patients because of poor predefined feasibility. A total of 98 patients were recruited to the gemcitabine/docetaxel arm (ie, gemcitabine followed by docetaxel). RESULTS: Quality of life remained near baseline levels during the administration of 6 cycles of gemcitabine/docetaxel chemotherapy, whereas it deteriorated after 2 cycles of docetaxel/gemcitabine. Toxicity was comparable between arms. Median times to progression were 4.3 months and 2.2 months with gemcitabine/docetaxel and docetaxel/gemcitabine, respectively, and median overall survival times were 9 months (gemcitabine/docetaxel) and 5 months (docetaxel/gemcitabine; P=0.029, Wilcoxon rank-sum test). CONCLUSION: These results indicate that first-line gemcitabine followed by second-line weekly docetaxel is feasible, with promising survival in patients with advanced NSCLC. PMID- 16354318 TI - Efficacy and safety of paclitaxel poliglumex as first-line chemotherapy in patients at high risk with advanced-stage non-small-cell lung cancer: results of a phase II study. AB - BACKGROUND: The objectives of this open-labeled, multicenter, phase II trial were to evaluate response, survival, and tolerability in patients at high risk (Eastern Cooperative Oncology Group performance status [PS] of 2 or age >or= 70 years) with advanced-stage non-small-cell lung cancer (NSCLC) receiving single agent paclitaxel poliglumex as first-line monotherapy. PATIENTS AND METHODS: Paclitaxel poliglumex was administered as a 10-20-minute infusion on day 1 of each 3-week cycle. Thirty patients were enrolled: 28 received paclitaxel poliglumex 175 mg/m2, and 2 received 235 mg/m2. Patients exhibiting a partial response (PR; by Response Evaluation Criteria in Solid Tumors) or stable disease (SD) continued uninterrupted treatment with paclitaxel poliglumex for or= 70 years was 7.8 months. No grade 4 nonhematologic toxicities were reported. Three patients experienced grade 3 neuropathy. Weekly hematologic assessments showed grade 3 anemia in 2 patients, grade 3 neutropenia (not associated with neutropenic fever) in 3 patients, and grade 4 neutropenia in 2 patients. No patient required growth factor support. CONCLUSION: The results of this study indicate that paclitaxel poliglumex is generally well tolerated and has activity at a dose level of 175 mg/m2 as first line monotherapy in patients at high risk with advanced NSCLC. PMID- 16354319 TI - The role of mitogen-activated ERK-kinase inhibitors in lung cancer therapy. AB - The Ras-Raf-MAPK pathway has been implicated in lung carcinogenesis and, potentially, the maintenance of the malignant phenotype in these tumors. Mutations in ras and B-raf genes have been described in lung cancer, representing one of the few examples of tandem mutations in a signaling cascade. As a result, numerous approaches to inhibiting this pathway in lung cancer have been explored in the past decade. The most promising approach to date appears to be the inhibition of mitogen-activated ERK kinase or MEK. In this review, the potential utility of MEK inhibitors in the therapy of lung cancer is discussed. PMID- 16354320 TI - Meeting highlights from 2nd symposium on controversies and clinical challenges in myeloma, lymphoma and leukemia. PMID- 16354323 TI - New approaches to treatment for multiple myeloma: durable remission and quality of life as primary goals. AB - New treatment approaches are changing the traditional paradigm for myeloma management. Partial or complete response with first-line therapy is now highly attainable. The focus of attention has thus shifted to obtaining the most durable remissions with the highest quality of life. A critical open question is whether more arduous and toxic therapies are justified with the intent to seek a cure. Patients with minimal symptoms at diagnosis are particularly reluctant to pursue aggressive strategies without documented long-term benefit. Conversely, patients with poor-risk molecular features, such as 13q deletion or t(4;14) translocation, can hopefully benefit from novel targeted therapies. New combinations incorporating bortezomib, thalidomide, and/or lenalidomide plus other novel agents offer the opportunity to explore therapy that is more effective and less toxic than in the past. The efficacy of single and tandem transplantation, which have documented long-term survival benefit, need to be compared with regimens integrating novel therapies. Whether true complete remission is a prerequisite for substantially improved survival is a central question in the framework of planned trials. The ultimate goal is to achieve clinical response (complete or partial) that offers the best quality remission for the longest period. Ideally, more effective induction and/or consolidation treatments will avoid concomitant toxicities and the need for maintenance therapies. Fortunately, new agents already offer longer-term disease control. The ongoing search for a cure will undoubtedly demand courage and dedication on the part of investigators and patients. PMID- 16354324 TI - Marked clinical activity of the proteasome inhibitor bortezomib in patients with follicular and mantle-cell lymphoma. AB - Recent advanced developments in our understanding of cancer cell biology have begun to generate a host of new targets that are proving to be valuable substrates for new drug development. One example includes our ever-increasing understanding of the complex biology surrounding the ubiquitin-proteasome pathway. For years there have been a variety of compounds used in the laboratory that have been shown to inhibit the proteasome, though many of these compounds have proven to be relative non-specific inhibitors of intracellular and proteasome proteases. The recent synthesis of 1 novel inhibitor, bortezomib (formerly known as PS341), has proven to be an effective reversible inhibitor of the chymotryptic protease in the 26S proteasome. Proteasome inhibition represents a new approach for the treatment of many forms of cancer, especially select hematologic malignancies. Bortezomib has been approved by the United States Food and Drug Administration for the treatment of relapsed or refractory multiple myeloma. In addition to myeloma, bortezomib has also shown promising activity in the treatment of select types of non-Hodgkin's lymphomas (NHLs). Several single agent phase II clinical trials in patients with a host of different NHL histologies have demonstrated that bortezomib has reproducible activity in mantle cell lymphoma (MCL) and follicular lymphoma (FL), with some suggestion of activity in marginal zone lymphoma. The promising activity in these smaller studies has led to a number of larger multicenter studies with bortezomib in combination with rituximab in MCL, FL, and marginal zone lymphoma. The collective early experience from these studies continues to support the activity of bortezomib in these histologies of NHL. Herein, some of the biologic rationale for using proteasome inhibitors in lymphoma as well as some of the clinical data from these promising studies are discussed. PMID- 16354325 TI - Risk stratification of patients with newly diagnosed multiple myeloma: optimizing treatment based on pretreatment characteristics. AB - Marked clinical and biologic heterogeneity exists in multiple myeloma (MM). Over the years, many prognostic factors have been identified and several prognostic systems have been proposed. The integration of data from international groups including patients treated with common modalities such as chemotherapy and high dose therapy culminated in the International Staging System. Recently, genetic information has also been shown to include powerful prognostic factors across different treatment modalities. These advances have facilitated categorization of patients into different risk groups, particularly a subset of patients at high risk with short survival times after current standard therapy. The expanding armamentarium of effective treatments in MM also means that it is now possible to select treatments for patients based on their risk categories. This review will summarize the important prognostic factors identified to date, how they can be used to identify patients at high risk, and their clinical utility in relation to treatment optimization at diagnosis. PMID- 16354326 TI - Amyloidosis: diagnosis and management. AB - Amyloidosis is a rare plasma cell proliferative disorder. The annual incidence in Olmsted County, Minnesota, is 8 in 1,000,000 patients. This is a difficult disorder to diagnose, because the symptoms at presentation are vague and include dyspnea, paresthesias, edema, weight loss, and fatigue. The clinical syndromes at the time of presentation include nephrotic-range proteinuria with or without renal failure, cardiomyopathy, "atypical multiple myeloma," hepatomegaly, and autonomic or peripheral neuropathy. The serum immunoglobulin free light chain assay has been an important step forward in classifying systemic amyloidosis as an immunoglobulin light chain form and in monitoring therapy. Recently, the importance of serum cardiac biomarkers in assessing outcome has been recognized. New therapies developed over the past 5 years include high-dose chemotherapy with stem cell reconstitution, combinations of alkylating agents with dexamethasone, and, most recently, thalidomide. PMID- 16354327 TI - Primary pulmonary lymphoma: current status. AB - Primary pulmonary lymphoma (PPL) is a rare disease with a favorable prognosis compared with lung cancer. Although a number of histologic variables of B- and T cell lymphoma were reported as PPL, marginal zone lymphoma of the mucosa associated lymphoid tissue type is by far the most frequent diagnosis. This review summarizes the present knowledge of histopathology, molecular biology, diagnosis, prognosis, and treatment of this heterogeneous entity. PMID- 16354328 TI - Advances in multiple myeloma and spine disease. AB - Progressive bone destruction is the hallmark of multiple myeloma (MM) and is responsible for principal morbidity in the disease. The spine is the most afflicted skeletal organ, and vertebral fractures have significantly contributed to its poor prognosis. The principal underlying pathologic mechanism causing bone disease in MM is a shift in the balance of bone formation and bone resorption toward bone resorption, and eventually total dissociation between the 2 processes occurs in latter stages of the disease. During the past decade bisphosphonates have become an important adjunctive treatment in the management of MM, in which they have shown the ability to reduce bony complications associated with the disease. Advances in minimally invasive surgical techniques, such as percutaneous vertebroplasty and kyphoplasty, offer these patients less-invasive options for the treatment of vertebral collapse and restoration of their normal function. This report reviews recent advances in the understanding of bone disease in MM, the role of bisphosphonates in the prevention of skeletal events, and available data regarding percutaneous vertebroplasty and kyphoplasty. PMID- 16354329 TI - T-cell prolymphocytic leukemia: a single-institution experience. AB - BACKGROUND: T-cell prolymphocytic leukemia is an uncommon, aggressive, mature T cell leukemia characterized by proliferation of T-cell lymphocytes. The recent availability of modern immunophenotypic and molecular tools has allowed a better distinction of this disorder from its B-cell counterpart and other mature T-cell leukemias. PATIENTS AND METHODS: The clinical, pathologic, and cytogenetic features of 57 patients with T-PLL who were evaluated at the Department of Leukemia, M. D. Anderson Cancer Center (MDACC) from 1986 to 2004 were examined. RESULTS: The most common cytogenetic abnormality was inv(14)(q11;q32), which was present in 7 patients. In all 7 patients, this abnormality was associated with other chromosomal aberrations. Patients treated with alemtuzumab at MDACC had a significantly better response rate (P = 0.02) and survival rate (P = 0.002). There were no significant differences in survival based on Tcl-1 expression or different patterns of CD4 and CD8 expression. CONCLUSION: Treatment with alemtuzumab results in higher response rates and a better survival rate in patients with T-cell prolymphocytic leukemia. PMID- 16354330 TI - Treatment and survival of patients with thyroid lymphoma: a population-based study with clinical and pathologic reviews. AB - PURPOSE: The purpose of this study was to determine the incidence, clinical and histologic features, and patterns of outcome of thyroid lymphomas. PATIENTS AND METHODS: A retrospective population-based survey of 38 patients with thyroid lymphoma was taken. Median age was 69 years (range, 33-87 years), with a 1:4 female predominance. Fifty percent of cases had a history of autoimmune thyroiditis, and coexistent thyroiditis was found in 67% of cases in which preexistent thyroid tissue was present. The most common subtype was diffuse large B-cell lymphoma (DLBCL; 63%) followed by extranodal marginal zone lymphoma (ENMZL; 29%). Ten of the patients with DLBCL showed a concomitant low-grade mucosa-associated lymphatic tissue component, and 4 cases of aggressive ENMZL were diagnosed. At diagnosis, 22 patients (58%) had localized disease, and 41% had low-risk international prognostic index scores. RESULTS: Therapy was diverse and included all possible treatment modalities, none of which showed superiority. A complete clinical response was exhibited in 64% of patients, 14% exhibited a partial response, and 22% developed progressive disease. At a median follow-up of 43 months (range, 0-240 months), 15 patients had relapsed or developed progressive disease. Two-year overall survival rate was 59% for all patients, 68% for patients with localized disease, and 47% for patients with disseminated lymphoma. CONCLUSION: Many thyroid lymphomas have clinical and histologic features characteristic of ENMZL and belong to this specific clinicopathologic entity. PMID- 16354332 TI - Activation-induced cytidine deaminase and CD38 expression in B-cell chronic lymphocytic leukemia. PMID- 16354331 TI - Fatal West Nile Virus infection after rituximab/fludarabine--induced remission for non-Hodgkin's lymphoma. AB - West Nile virus (WNV) infections are potentially life threatening in immunocompromised hosts. Currently, the best diagnostic test is serology. Reverse transcriptase polymerase chain reaction (RT-PCR) testing has a role, but, because WNV is a cell-associated neurotropic virus, RT-PCR results are frequently negative even in cases of active infection. We present a case in which serology results were persistently negative because the patient was immunocompromised following lymphoma treatment. The role of humoral immunity in resolution of WNV is also discussed. PMID- 16354334 TI - The aetiology and management of atrophic rhinitis. AB - Atrophic rhinitis is a chronic, debilitating and recalcitrant disease of the nasal cavities that is prevalent in several parts of the world. It has unique epidemiological features and clinical characteristics. Clinicians and researchers for decades have tried to postulate theories for the aetiology of the primary form of the disease. Management of the disease has seen several medical therapeutic regimens including alternative forms of medicine. Surgical options for the condition are also not completely satisfactory with a number of failures and recurrences. The authors provide here a comprehensive review of the existing literature as regards the aetiology and management of this refractory condition. PMID- 16354335 TI - Epilepsy following middle-fossa extradural retraction: implications for driving. AB - New-onset epilepsy has several social and financial repercussions. Development of seizures after middle-fossa surgery is uncommon. We present two subjects who developed epilepsy following middle-fossa surgery, requiring treatment with anticonvulsants, and discuss the implications. PMID- 16354336 TI - A multipurpose landmark for skull-base surgery: Henle's spine. AB - OBJECTIVE: To determine whether Henle's spine could be used as a reliable and multipurpose landmark for the other important structures of the skull base. MATERIALS AND METHODS: Ninety-two specimens from 46 cadaveric adult dry skulls were studied. Two imaginary lines and a triangle were defined: a spinopterygoidal line extending from Henle's spine to the root of the medial pterygoid plate, a bispinal line extending from one Henle's spine to the contralateral one, and a parapetrosal triangle lying between the spinopterygoidal line, the bispinal line and the sagittal midline. The parapetrosal triangle encompasses nearly all the main structures of the skull base, including the petrosal internal carotid artery. RESULTS: Along the spinopterygoidal line the distance from Henle's spine to the spine of the sphenoid was found to be about 3 cm, to the foramen spinosum 3.5 cm, to the posterior and anterior margins of the foramen ovale 4 and 4.5 cm, to the root of the lateral pterygoid plate 5 cm, to the root of the medial pterygoid plate 5.5 cm, and to the vomer 6.5-7 cm. Along the bispinal line, the distance from Henle's spine to the stylomastoid foramen was found to be about 1.5 cm, to the lateral and medial margins of the jugular foramen 2.5 and 3.5 cm, to the external orifice of the hypoglossal canal 4 cm, and to the foramen magnum 5 cm. CONCLUSION: Henle's spine with its superficial and central position can be used to localize important anatomical structures during skull-base surgery. PMID- 16354337 TI - Impact of antibiotics on pathogens associated with otitis media with effusion. AB - OBJECTIVE: To further understand the roles of bacteria and antibiotics in the development of otitis media with effusion (OME). METHODS: Samples of middle-ear effusion (MEE) were collected during the placement of ventilation tubes to treat chronic OME. Children with acute otitis media within the past three months were excluded from this study. We used polymerase chain reaction (PCR) to detect pathogens and to test the susceptibility of Streptococcus pneumoniae to penicillin. RESULTS: Among MEE samples from 52 children, PCR detected bacterial DNA in 32 per cent (24/75) of them. S. pneumoniae was detected more frequently in middle ears that required ventilation tube insertion at least twice compared with those requiring ventilation tube insertion only once (5/15 versus 4/60; p = 0.013). Higher levels of S. pneumoniae were detected in MEE from children with, than without, a long history of antibiotic administration (7/10 versus 2/14; p = 0.0187). The pbp genes of all isolated S. pneumoniae contained mutations. CONCLUSIONS: Long exposure to antibiotics might significantly influence the bacterial genome in MEE. PMID- 16354338 TI - Endonasal endoscopic repair of anterior skull-base fistulas: the Kuala Lumpur experience. AB - The purpose of this retrospective study is to determine the pattern of cerebrospinal fluid (CSF) rhinorrhoea presenting to our tertiary referral centre in Kuala Lumpur and to assess the clinical outcomes of endonasal endoscopic surgery for repair of anterior skull base fistulas. Sixteen patients were treated between 1998 and 2004. The aetiology of the condition was spontaneous in seven and acquired in nine patients. In the acquired category, three patients had accidental trauma and this was iatrogenic in six patients (five post pituitary surgery), with one post endoscopic sinus surgery (ESS). Imaging included computed tomography (CT) scan and magnetic resonance imaging (MRI). Endoscopic repair is less suited for defects in the frontal sinuses with prominent lateral extension and defects greater than 1.5 cm in diameter involving the skull base. Fascia lata, middle turbinate mucosa, nasal perichondrium and ear fat ('bath plug') were the preferred repair materials in the anterior skull base, whereas fascia lata, cartilage and abdominal fat obliteration was preferentially used in the sphenoid leak repair. Intrathecal sodium flourescein helped to confirm the site of CSF fistula in 81.3 per cent of the patients. Ninety per cent of the patients who underwent 'bath plug' repair were successful. The overall success rate for a primary endoscopic procedure was 87.5 per cent, although in two cases a second endoscopic procedure was required for closure. In the majority of cases endoscopic repair was successful, and this avoids many of the complications associated with craniotomy, particularly in a young population. Therefore it is our preferred option, but an alternative procedure should be utilized should this prove necessary. PMID- 16354339 TI - Allergic fungal sinusitis (AFS)--earlier diagnosis and management. AB - AIM: To evaluate the criteria for diagnosing allergic fungal sinusitis and to maintain permanent drainage and ventilation, while preserving the integrity of mucosa. METHODS: This is a prospective study of 251 patients with chronic rhinosinusitis with or without polyposis, of whom 199 were treated surgically. Mucus sample collection, nasal secretion culture, surgical specimen handling and histological evaluation of surgical specimens are described. The management included wide local endoscopic sinus debridement, adequate sinus aeration, post operative use of steroids and antifungal therapy. RESULTS: Fungal cultures of nasal secretions were positive in 201 (80.01 per cent) of 251 patients. Of the 199 surgical cases, fungal elements were found in 156 histological specimens (62.1 per cent). Allergic mucin was found in 182 patients (91.45 per cent). Nasal obstruction and proptosis were the commonest presentations. All pre-operative versus post-operative changes in AFS-associated complaints reached statistical significance of p < 0.001. The ethmoid sinus was commonly involved with adjacent lamina papyracea exhibiting demineralization in 26.6 per cent of cases. Intracranial extension was seen in 15 cases. Recurrence was noted in 11 cases. CONCLUSION: Comprehensive treatment with endoscopic sinus surgery, steroids and antifungal therapy is needed. AFS is readily recurrent. Long-term follow up is important. PMID- 16354340 TI - Usefulness of technetium-99m methoxyisobutylisonitrile single-photon emission computed tomography and computed tomography in the evaluation of cervical lymph node metastasis. AB - The aim of this study was to compare the effectiveness of technetium-99m methoxyisobutylisonitrile ((99m)Tc-MIBI) single-photon emission computed tomography (SPECT) and computed tomography (CT) in evaluating cervical lymph node metastasis of head and neck cancer. Histopathologic results of 166 cervical lymph node levels in 31 neck-dissected patients were compared with pre-operative CT and (99m)Tc-MIBI SPECT findings about cervical lymph node metastasis, retrospectively. Sensitivity, specificity and predictability of CT and (99m)Tc MIBI SPECT were 68.2, 93.1 and 89.8 per cent and 59.1, 87.5 and 83.7 per cent, respectively. When analysing CT and(99m)Tc-MIBI SPECT together, sensitivity and specificity were 86.4 and 99.3 per cent, respectively. The combined use of (99m)Tc-MIBI SPECT and CT could increase the accuracy of cervical lymph node metastases detection, compared with separate use of either (99m)Tc-MIBI SPECT or CT. PMID- 16354341 TI - Radiofrequency vs laser in the management of mild to moderate obstructive sleep apnoea: does the number of treatment sessions matter? AB - OBJECTIVE AND HYPOTHESIS: Obstructive sleep apnea (OSA) is a relatively common and serious problem with many medical and social consequences. Laser and radiofrequency are two recent techniques used to treat OSA and they can be carried out under local anaesthesia, but they need multiple sessions to achieve satisfactory outcome and are associated with better short-term than long-term outcomes. In this work we compare the two modalities as regards the optimal number of treatment sessions needed to achieve a favourable outcome in the short and long term. STUDY DESIGN: A total of 150 patients with apnoea hypopnoea index (AHI) between 5 and 30 events per hour, no morbid obesity and retropalatal site of obstruction were included in this prospective study. METHODS: Patients were randomly and equally divided into two groups, each comprising 75 patients. The first group was treated with bipolar radiofrequency volumetric tissue reduction of the palate (BRVTR) and the second group was treated with laser-assisted uvulopalatoplasty (LAUP). Each group was further subdivided into five subgroups each consisting of 15 patients. The first group received one treatment session, the second received two sessions, the third received three sessions, the fourth received four sessions and the fifth group received five treatment sessions. Evaluation of efficiency of both techniques in treating OSA was assessed objectively by polysomnography. RESULTS: In those treated with BRVTR; at least three sessions were needed to achieve a favourable outcome in OSA in the short and long term. In those treated with LAUP, a single treatment session was enough to achieve a favourable outcome on OSA in the short term, while two sessions were needed to achieve the same long-term outcome. DISCUSSION: In OSA, fewer treatment sessions are needed with LAUP (one session) than with BRVTR (three sessions) to achieve a favourable outcome. In LAUP more treatment sessions (two) are needed to maintain a longer-term favourable outcome than those needed to achieve short-term favourable outcome (one session), which is not the case with BRVTR (three sessions are needed to achieve both short- and long-term favourable results). PMID- 16354342 TI - The influence of weather on the frequency of secondary post-tonsillectomy haemorrhage. AB - The aim of this study was to determine whether certain weather variables influence the secondary post-tonsillectomy haemorrhage rate and to examine the influence of a change in these variables on secondary haemorrhage. This was a prospective study carried out in a tertiary referral institution. All patients undergoing bilateral tonsillectomy over a one-year period were included. Local weather data, including daily temperature (max/min), relative humidity and water vapour pressure, were acquired. All patients readmitted to our department with secondary post-tonsillectomy haemorrhage were recorded. Of 346 patients undergoing tonsillectomy, 32 developed secondary haemorrhage (9.2 per cent). A significant negative correlation was found between secondary haemorrhage rate and the average monthly temperature (max/min) and water vapour pressure (Pearson's correlation = -0.8) (p < or = 0.002). These results suggest that performing tonsillectomy in warmer weather when the water vapour pressure is higher may reduce the secondary haemorrhage rate. PMID- 16354343 TI - Videoendoscopic surgery for inaccessible glottic lesions. AB - OBJECTIVES: Lesions in the anterior segment of the vocal fold are sometimes difficult to access by means of conventional suspension microlaryngoscopy under general anaesthesia because of anatomical factors such as short, stout and inflexible necks, reduced jaw protrusion, and long incisors. Various techniques have recently been proposed for the management of inaccessible glottic lesions, most of which are performed under general anaesthesia. The use of flexible videoendoscopic surgery under topical anaesthesia in two cases of anterior glottic lesions that could not be treated by means of conventional suspension laryngoscopy is described. STUDY DESIGN: Case report. METHODS: A flexible videobronchoscope with an instrument channel was inserted transnasally on an out patient basis. While the examiner carried out the endoscopy, an assistant maneuvered the biopsy forceps through the instrument channel, and removed the lesion. RESULTS: Both patients underwent successful removal of an anterior glottic polyp, and the one-year follow-up evaluation revealed normal anatomy of the vocal folds and normal vocal function. CONCLUSIONS: Flexible videoendoscopic surgery under topical anaesthesia is a safe, simple and minimally invasive procedure that can be considered as an alternative to traditional endoscopic surgery for inaccessible anterior glottic lesions. PMID- 16354344 TI - Mediastinal tracheostomy: unilateral resection of the anterior chest wall. AB - Since the first reliable mediastinal tracheostomy described by Grillo et al. in 1966, many new techniques have been described in order to reduce the number of complications. We here report the case of a 55-year-old man who was referred for surgery with post-radiochemotherapy recurrence of a double neoplasm of the pharyngolarynx extending to the proximal trachea and the medial part of the oesophagus. Through a median sternotomy, a pharyngolaryngoesophagectomy was performed with an extended tracheal resection. The reconstruction of the upper digestive tract was performed with a gastric pull-up. The mediastinal tracheostomy was performed with a pectoralis major muscular flap through a right unilateral resection of the manubrium, the right clavicular head and the right first and second costal cartilages. Historically, the mediastinal tracheostomy was performed through a large bilateral resection of the anterior chest wall, in order to prevent the tension on the tracheocutaneous sutures. Nowadays, with the possibility of various pedicled flaps, bilateral resection no longer seems to be necessary. This unilateral resection leads to a reduction in post-operative sequelae. PMID- 16354345 TI - Upper neck spinal accessory nerve identification during neck dissection. AB - Iatrogenic injury to the spinal accessory nerve (SAN) during neck dissection may result in significant and avoidable morbidity in the form of 'shoulder syndrome'. The authors describe a simple method, based on the anatomy of the sternocleidomastoid muscle (SCM), which allows consistent and rapid identification of the SAN in the upper neck during dissection, thereby facilitating its preservation. PMID- 16354346 TI - Survival of a patient with endopharyngeal common carotid blow out due to parapharyngeal abscess as an unusual complication of cholesteatoma. AB - Parapharyngeal abscess as a complication of cholesteatoma is an uncommon entity. Endopharyngeal common carotid artery rupture due to parapharyngeal abscess is also uncommon, and these cases usually end fatally. We present a 17-year-old male with parapharyngeal abscess due to cholesteatoma who developed an endopharyngeal common carotid blow out and survived after common carotid ligation without any neurological sequalae. PMID- 16354347 TI - Pharyngeal perforation: a late complication of cervical spine surgery. AB - Anterior cervical spine fusion and stabilization is a well-recognized procedure for a number of cervical spine disorders. Unfortunately, the complex anatomy of the cervical spine means that these procedures are not without complications. Pharyngo-oesophageal perforation is a rare but potentially life-threatening complication of cervical spine surgery and may present intra-operatively, in the immediate post-operative period or many years later. We present the case of a gentleman with ankylosing spondylitis who presented with a pharyngeal perforation and fistula five years after cervical spine surgery. PMID- 16354348 TI - Telomerase catalytic subunit gene expression does not influence survival of patients with squamous cell carcinoma of the larynx and hypopharynx: a case control study. AB - AIMS: to determine correlations between relative quantities of telomerase catalytic subunit m-ribonucleic acid (hTERT mRNA) and conventional clinicopathological parameters (such as site, size and grade of tumour, the presence of regional lymph node metastases, and, in particular, survival) in patients with laryngeal and hypopharyngeal squamous cell carcinomas (SCCs). MATERIAL AND METHODS: The relative quantity of hTERT mRNA was analysed by a commercially available LightCycler Telo TAGGG hTERT Quantification Kit in 56 cases of SCC (40 laryngeal and 16 hypopharyngeal). The association with cancer specific survival was evaluated by univariate and multivariate analysis. RESULTS: Location of the tumour in the hypopharynx was the only significant negative predictive factor for survival, as determined by univariate analysis (p = 0.028). Although a tendency towards a better overall survival was observed for female patients younger than 50 years, for lower tumour grades and sizes, and for the absence of regional lymph node metastases, the prognostic significance of these factors could not be confirmed. No differences existed in hTERT mRNA expression between laryngeal and hypopharyngeal SCCs. Furthermore, no correlation was found between the relative quantities of hTERT mRNA and the tumour size, regional lymph node metastases or survival of patients with laryngeal or hypopharyngeal SCCs. CONCLUSIONS: The results of the present study suggest that genetic abnormalities other than telomerase reactivation are responsible for progression of laryngeal and hypopharyngeal SCCs. PMID- 16354349 TI - Late laryngeal radionecrosis in severe arteriosclerosis. AB - Laryngeal necrosis is a serious complication that usually occurs within the first year following completion of radiotherapy, although it is reported that cases can develop after a long period of latency. Factors such as dosage and irradiation technique employed, tumour invasion into the laryngeal cartilage, infection, continued smoking, trauma and general vascular condition of the patient have been considered to increase the rate and degree of development of radionecrosis. We report an unusual case of laryngeal radionecrosis in a patient with hypertension, hyperlipidaemia, diabetes and a history of cigarette smoking, which developed 25 years after radiotherapy for laryngeal carcinoma. His systemic illnesses and continued smoking were speculated to have contributed to the progress of the radionecrosis, suggesting that cessation of smoking and control of arteriosclerotic diseases should be considered to decrease its incidence. PMID- 16354350 TI - Spitting in the ear: a falsified disease using video evidence. AB - We present the case of Munchausen's syndrome by proxy (MSBP) in which the mother spat and placed blood in her child's ear, and videoed the result, in order to gain the attentions of the medical profession. This is the first case report of this kind and may represent a disturbing trend in the use of digital photography and video to support factitious illnesses. PMID- 16354351 TI - Ectopic submandibular gland presenting as a swelling in the floor of mouth. AB - The authors describe a patient who presented with a left sublingual swelling. Ultrasonic and sialographic investigation showed no salivary tissue in the expected left submandibular region, but a large salivary gland in the left floor of the mouth deep to the mylohyoid muscle. This is likely to represent an ectopically positioned submandibular gland. PMID- 16354354 TI - Expression of female sex hormone receptors in human middle-ear cholesteatomas. AB - The purpose of this study was to establish the eventual presence of progesterone receptor (PGR) and oestrogen receptor (EGR) in human middle-ear cholesteatoma (MECh) tissues and to compare their expression between male and female patients. An immunohistochemical technique was employed for detection of PGR- and EGR specific immunoreactivity in MECh samples using formalin-fixed paraffin-embedded tissue sections. The positive results were verified with reverse transcriptase polymerase chain reaction (RT-PCR). The morphological study revealed stable expression of PGR in suprabasal layers of all cholesteatoma samples. Weaker immunoreactivity for PGR was demonstrated in external auditory canal skin (EACS) samples in comparison with MECh, while PGR-specific staining was not observed in retroauricular skin (RAS) samples. EGR was detected only at mRNA levels. Stronger expression of EGR PCR products was disclosed in female cholesteatoma samples, while PGR mRNA was predominantly detected in male cholesteatoma specimens. Our preliminary experimental results give us ground to assume that female sex hormones may stimulate proliferation and affect differentiation of MECh keratinocytes. PMID- 16354355 TI - Cochlear implantation in chronic otitis media. AB - Chronic otitis media (COM) patients who had a multichannel device implanted were evaluated regarding surgical problems and technical modifications. In a multicentric study, implantees whose aetiology was COM were retrospectively evaluated. Patients were operated on and evaluated at three different tertiary referral centres: SSK Izmir Hospital Cochlear Implantation (CI) Center (32 cases), Istanbul Marmara University ENT Clinic (six cases), Eskisehir Anadolu University CI Center (one case). Thirty-nine implantees were evaluated with respect to surgical problems, technical modifications, complications and hearing results. All patients had lost their hearings as a result of COM. Three out of 39 cases were children. Thirty-seven of the patients either had a radical cavity or ear converted to radical at the first stage or concomittantly with the implantation. Patients were evaluated in order to find out the best possible surgical solutions to specific problems caused by COM. In seven cases electrode array disrupted the epithelial lining of the cavity despite specific measures. Five of those cases were re-implanted, passing the electrode array through a tunnel under the facial nerve. Seven recent cases were also implanted with the same subfacial route. In all patients but one, satisfactory hearing results were achieved one to five years after implantation (SDS scores with monosyllabic word list were between 59 and 89 per cent, median 67.4 per cent). CI in COM patients necessitates technical modifications. In radical cavities subfacial implantation seems to be a good solution for the protection of the electrode array. PMID- 16354356 TI - A comparative study between ciliary count and the degree of opacity of paranasal sinus CT scans in chronic sinusitis pre and post FESS. AB - OBJECTIVE: To investigate the relationship between the extent of sinus disease in chronic sinusitis as detected radiologically by computed tomography (CT) scan and the population of cilia (ciliary area) both before and after functional endoscopic sinus surgery (FESS). In a simple way this is a trial to statistically prove that the CT scan could be a valid indicator and a mirror of the histological status of the sinus mucosa. DESIGN: Twenty adult patients were enrolled in this study. Radiological extension of the sinus disease was quantitated using the classification proposed by Kennedy in 1992 and the ciliary population was studied using scanning electron microscopy and image analysis softwares. RESULTS AND CONCLUSION: The more advanced the sinusitis, as evidenced by CT scans, the more the expected reduction in the ciliary area (CA) and in the ciliary count. But after FESS the degree of ciliary regeneration does not depend statistically on the radiological condition of the sinuses and the degree of opacity prior to intervention, i.e. a statistically valid inverse relationship exists between the radiological stage of sinusitis and the ciliary population pre operatively but the same relation does not extend to the ciliary population post operatively. PMID- 16354357 TI - Passive smoking, allergic rhinitis and nasal obstruction in children. AB - Allergic rhinitis is a common cause of nasal obstruction in childhood. This prospective study looked at the effect of passive smoking on nasal obstruction in children with and without allergic rhinitis. Eighty-one children took part. Each child was asked to score his or her degree of nasal obstruction on a visual analogue scale. Exposure to passive smoking was determined subjectively using a parental questionnaire, and objectively by measuring the urinary cotinine/creatinine ratio. Results were tabulated using Microsoft Excel and analysed with SPSS statistical software. Nasal obstruction was significantly worse in children with a positive history of allergic rhinitis (p < 0.05). There was also a trend towards a higher nasal obstruction score in children without allergic rhinitis exposed to passive smoking compared to those who were not so exposed. As would be expected, nasal obstruction is worse in children with allergic rhinitis than in those without. Passive smoking tends to increase the symptom of nasal obstruction in children without allergic rhinitis. PMID- 16354358 TI - Aryepiglottoplasty for laryngomalacia: the Alder Hey experience. AB - Laryngomalacia is the most common cause of stridor in infants. Severely affected children are at risk of feeding difficulties, apnoeic episodes and cor pulmonale secondary to upper airway obstruction. The aim of this study was to assess the outcome of aryepiglottoplasty. This is a simple surgical procedure that relieves the obstruction by dividing the aryepiglottic folds. Thirty children had an aryepiglottoplasty at the Royal Liverpool Children's Hospital between January 1995 and June 2001. The case notes of all 30 children were reviewed for age, sex, age at operation, indications, operative technique, complications and long-term outcomes. Complete resolution of stridor was obtained in 83 per cent of patients, with an improvement in a further 7 per cent. Post-operative complications included lower respiratory tract infections (13 per cent) and vomiting (7 per cent). In conclusion, simple endoscopic aryepiglottoplasty remains an effective way of treating upper airway obstruction in children. Its high resolution and low complication rate make it a safe, first choice procedure for treatment of moderate to severe laryngomalacia. PMID- 16354359 TI - Radiophonosurgery of benign superficial vocal fold lesions. AB - OBJECTIVE: To conduct a prospective randomized controlled trial describing and investigating the efficacy and safety of radiosurgical excision of benign superficial vocal fold lesions. MATERIALS AND METHODS: Fifty patients with benign superficial vocal fold lesions (20 vocal nodules, 27 vocal polyps and three Reinke's oedema) who failed conservative therapy were included in the study. They were equally randomized into cold knife or radiosurgical excision. Clinical and voice assessments were done pre-operatively and after surgery. Voice analysis included a subjective visual analogue scale (VAS) and a perceptual assessment with a simplified version of the GRBAS scale (GRB) consisting of G (grade), R (roughness), and B (breathiness). Acoustic voice evaluation included jitter and shimmer. Post-operative voice therapy was provided for all patients. Complications, smoothness of post-operative recovery, and administration of analgesia were reported. RESULTS: Both groups experienced significant improvement in VAS, perceptual evaluation and acoustic analysis after surgery, with no evidence of significant differences between the cold knife and radiofrequency groups. The radiofrequency group showed a decrease of 17 per cent in the mean operative time when compared with the cold knife group, but this was not statistically significant. No evidence of a significant difference was noticed in the smoothness of post-operative recovery, administration of analgesia and complication rate. CONCLUSION: Radiophonosurgery opens a new therapeutic approach for patients with benign superficial vocal fold lesions. It combines the advantages of both cold knife and laser phonosurgery, being easy, safe, precise and effective, and having excellent tactile and haemostatic properties. PMID- 16354360 TI - Acute epiglottitis: analysis of factors associated with airway intervention. AB - We reviewed acute epiglottitis (AE) and identified factors associated with airway intervention. This report was a retrospective review of patients with AE and compared with factors associated with airway intervention. We reviewed 96 patients who were diagnosed with AE in our hospitals in Japan. Ninety-two (96 per cent) patients were adults, and no seasonal variation in the incidence of AE was encountered. Eight (8 per cent) patients had tracheostomy and endotracheal intubation had not been done. We found that symptoms of stridor and muffled voice, a rapid clinical course, and diabetes mellitus were the factors associated with airway intervention. Extremely severe swelling of the epiglottis such that only less than half of the posterior vocal fold (scope classification (SC): III) could be seen, and extension of the swelling to the arytenoids (SC: B) were the two factors that were strongly associated with airway intervention. PMID- 16354361 TI - Thyroidectomy does not cause globus pattern symptoms. AB - Post-thyroidectomy patients often complain of globus pharyngeus type symptoms. One of the organic pathologies recognized as causing globus pattern symptoms is goitre. We conducted a prospective questionnaire-based study in the thyroid clinic at Hull Royal Infirmary to examine the relationship between goitre, thyroidectomy and globus pattern symptoms by using the Glasgow-Edinburgh Throat Scale. A questionnaire-based study in which 41 consecutive patients due to undergo thyroidectomy prospectively completed the questionnaire pre-operatively and 12 months post-operatively. A subset of 25 randomly selected patients also completed the questionnaire three months post-operatively. Globus symptoms were not worsened by thyroidectomy at three or 12 months. Indeed two of the questioned parameters were significantly improved at three months, and six at one year (p < 0.023). In conclusion, thyroidectomy does not exacerbate globus pharyngeus symptoms. Indeed, in the absence of other clinical causes, when a patient has a World Health Organization (WHO) 2 goitre or greater, thyroidectomy may improve them. PMID- 16354362 TI - Radiotherapy of the neck influences the distribution of laryngeal secretory glands. AB - To elucidate the influence of radiation therapy on the larynx in patients who receive radiotherapy to the neck, we observed the changes in the distribution of the laryngeal glands. Human adult larynges from patients with either laryngeal or hypopharyngeal cancer were histopathologically examined, and the changes in the glandular distribution as well as in the ratio of serous or mucous type cells were surveyed using an image analysis system. The mean ratio of the area occupied by glandular acini in the subglottic larynx was 0.182 in the non-radiation group and 0.098 in the radiation group (p < 0.001). Although no statistical difference was observed, the average density of the glands decreased and the average ratio of serous-type/mucous-type glandular cells decreased in the irradiated larynx. The glandular acini are often replaced with fibrous connective tissue and the ratio of serous-type glandular cells decreases. These results indicate that not only the voice function but also the local defence function of the larynx might be impaired after radiotherapy. PMID- 16354363 TI - The laryngeal dissection laboratory. AB - The purpose of this study was to dissect various animal larynges in support of a proposal for a laryngeal dissection laboratory. It was found that although the bovine larynx is larger than the human larynx, the anatomy is similar. Thirty three bovine larynges were dissected by the author in various simulated phonosurgical and partial laryngectomy procedures. Following each procedure, the specimen was photographed for teaching purposes. The author concludes that dissecting bovine larynges offers a valuable aid to training in the field of laryngeal surgery. It is intended to apply these dissections in the form of a departmental surgical workshop. PMID- 16354364 TI - Maturation sutures for the paediatric tracheostomy--an extra safety measure. AB - OBJECTIVE: Displacement of the tracheostomy tube in paediatric patients is a potentially fatal complication. We describe an extra safety measure which facilitates tube replacement. SETTING: Tertiary referral specialist paediatric centre. MATERIALS AND METHODS: The method involves the placement of sutures between the anterior tracheal wall and skin to hasten the formation of a mature stoma (maturation sutures). We also undertook a retrospective case note review on patients from an academic tertiary referral centre. Thirty-five notes were reviewed. The most common indication for tracheostomy was airway obstruction (65 per cent). Fourteen patients had early and 10 had late complications with three tube displacements occurring. No added complications due to the use of sutures were found. CONCLUSIONS: Our complication rates compare well with those in the literature, and we recommend considering the use of such a technique. PMID- 16354365 TI - Insertion of Provox2 voice prosthesis using the anterograde technique immediately after the secondary tracheoesophageal puncture. AB - Insertion of a voice prosthesis through the tracheoesophageal puncture (TEP) is one way to restore the voice after total laryngectomy. The second generation Provox voice prosthesis is presently one of the most popular devices. Although TEP can be done primarily, there are many centres that prefer it to be done at a second stage for various reasons. However, secondary TEP for retrograde replacement of prosthesis can be difficult and general anaesthesia is very often necessary. Moreover, the presence of neck stiffness and fibrosis from the surgery or previous radiotherapy could affect the neck extension for proper positioning of the trocar. Similarly, it is difficult to insert the prosthesis if there is stenosis at the pharyngoesophageal segment. We describe a technique in which creation of secondary TEP and insertion of Provox2 is done with local anaesthesia under the same setting. The procedure is well tolerated and can be safely performed on an out-patient basis. PMID- 16354366 TI - Clinical and prognostic aspects of laryngeal clear cell carcinoma. AB - Clear cell carcinoma (CCC) of the larynx is a rare pathological finding; only eight cases are described in the literature. It occurs in older adults and shows a predilection for men. We report the ninth observation of laryngeal CCC in the literature. We reviewed the literature and correlated the prognosis of the tumour according to its site of onset and treatment. The literature review showed that this neoplasm is highly aggressive, with a high recurrence rate and short mean survival time; the treatment of choice is surgery, and chemo- or radiotherapy are used mainly for the treatment of recurrences. PMID- 16354367 TI - Stenosis of the internal auditory meatus masquerading as bilateral vestibular schwannomas: a cautionary tale. AB - Stenotic malformations of the internal auditory meatus (IAM) are rare. They are known to symptomatically mimic vestibular schwannomas leading to potential diagnostic error. We present a case (along with literature review) where a stenotic IAM was clinically and radiologically misdiagnosed as a vestibular schwannoma. PMID- 16354368 TI - Pneumomediastinum due to tracheal foreign body granuloma. AB - Primary tracheal masses are rare. Secondary masses of the trachea are commonly foreign body granuloma, intubation granuloma or viral granuloma. The differential diagnoses given in such cases include both benign and malignant lesions. The otolaryngologist is often asked to perform a biopsy of the lesion to arrive at a diagnosis. However, even malignant processes can cause a granulomatous reaction. The timely diagnosis of tracheal masses depends upon maintaining a high index of suspicion. A rare case of tracheal granuloma leading to pneumomediastinum in a 53 year-old female is presented. The clinical features, investigations and treatment are detailed. The tracheal granuloma was managed by excision through bronchoscopy and the pneumomediastinum was managed conservatively. PMID- 16354369 TI - A large and uncharacteristically aggressive cholesterol granuloma of the middle ear. AB - Cholesterol granulomas of the middle ear tend to be benign in nature, frequently seen by otologists during cholesteatoma or chronic otitis media surgery and easily dealt with by evacuation. In contrast, cholesterol granulomas of the petrous apex appear to have a more aggressive nature and present significant pathological threats. We present a case of a large destructive cholesterol granuloma of the middle ear eroding the middle and posterior cranial fossa bone and dura to exert a mass effect upon the temporal lobe. We emphasize the destructive potential of this extremely rare middle-ear cholesterol granuloma, and draw attention to the distinction between this variant and the commonplace and benign form of the lesion more usually found at this site. We also present the current theories of the aetiology of cholesterol granuloma formation with possible explanations for the rare aggressive behaviour. PMID- 16354370 TI - Tubular apocrine adenoma with syringocystadenoma papilliferum arising from the external auditory canal. AB - Tubular apocrine adenoma (TAA) is a very rare sweat gland tumour. Comprehensive review of the literature reveals that TAA in the external auditory canal (EAC) has not yet been reported. We report a case of TAA in the EAC, together with characteristic histopathological findings. PMID- 16354371 TI - Pharyngeal perforation with deep neck abscess secondary to isolated hyoid bone fracture. AB - We report the case of a 26-year-old man who presented to us with dysphagia secondary to blunt trauma to the neck. The patient was found to have a hyoid bone fracture with pharyngeal perforation and resultant neck abscess. The patient responded to prompt surgical and medical management. We believe this to be the first report of such a case. PMID- 16354372 TI - Metastatic carcinoma mimicking a facial nerve schwannoma: the role of computerized tomography in diagnosis. AB - Secondary deposits in the temporal bone are uncommon but well recognized. Such tumours may involve the facial nerve by direct extension of the destructive process into the fallopian canal. We present a rare case of metastasis from a breast carcinoma in the facial nerve itself, involving the nerve in the internal acoustic meatus with extension into the labyrinthine segment, the first genu and into the middle-ear segment. The rest of the temporal bone was not involved. The lesion resembled a facial schwannoma on a routine magnetic resonance (MR) image. The diagnosis was confirmed after a post-operative computed tomography (CT) scan showed another separate secondary deposit in the basisphenoid. Histology was consistent with secondary tumour from a breast carcinoma. The case highlights the importance of keeping a high degree of suspicion for metastatic tumours in patients with a previous history of malignancy and the usefulness of CT scan in the evaluation of such cases. PMID- 16354373 TI - Diagnostic dilemma of an atraumatic clavicle fracture following radical treatment for laryngeal carcinoma. AB - An atraumatic clavicular fracture presented after radical treatment for laryngeal carcinoma. This presented a diagnostic dilemma. The differential diagnosis included metastatic bone disease and osteomyelitis as well as post-radiotherapy complications. After investigation, the cause was thought to be a post-radiation fracture of the clavicle and to the best knowledge of the authors, this is the first ever documented in a patient who had undergone a total laryngectomy with bilateral modified radical neck dissections and post-operative radiotherapy. Cases of a fractured clavicle post-radiation have been most commonly documented in patients with breast cancer and only a few cases have been documented in patients with laryngeal cancer treated with a total laryngectomy, bilateral radical neck dissections and radiotherapy. PMID- 16354374 TI - Leishmaniasis of the nasal cavity: a case report. AB - Establishing diagnosis of a granulomatous lesion of the nose is often difficult. Here we report a case of granulomatous lesion of the nose caused by Leishmania- an unlikely cause in the UK. The diagnosis and management of the case is discussed here. PMID- 16354375 TI - Dumbbell-shaped cervical spinal neurilemmoma presenting as neck mass. AB - Cervical neurilemmoma may originate from any nerve sheath tissue in the neck including the vagus nerve, glossopharyngeal nerve, brachial plexus, sympathetic trunk and cervical spine. We report an unusual case of a dumbbell-shaped neurilemmoma arising from the cervical spinal roots in a patient who complained of having had a neck mass for several months. Computed tomographic scan and magnetic resonance imaging revealed a dumbbell-shaped tumour extending from the C4 spinal level through the intervertebral foramen into the right parapharyngeal space. Decompression surgery was performed first via the cervical approach. Five months later, the patient received laminectomy and a complete tumour excision. The symptoms and signs were significantly relieved without neurological sequelae. No evidence of recurrence was noted after one-year follow up. This two-staged operation could offer an alternative surgical approach yielding ideal therapeutic results in such a rare disease. PMID- 16354376 TI - Importance of X-ray in chronic stridor. PMID- 16354378 TI - Sensory memory for odors is encoded in spontaneous correlated activity between olfactory glomeruli. AB - Sensory memory is a short-lived persistence of a sensory stimulus in the nervous system, such as iconic memory in the visual system. However, little is known about the mechanisms underlying olfactory sensory memory. We have therefore analyzed the effect of odor stimuli on the first odor-processing network in the honeybee brain, the antennal lobe, which corresponds to the vertebrate olfactory bulb. We stained output neurons with a calcium-sensitive dye and measured across glomerular patterns of spontaneous activity before and after a stimulus. Such a single-odor presentation changed the relative timing of spontaneous activity across glomeruli in accordance with Hebb's theory of learning. Moreover, during the first few minutes after odor presentation, correlations between the spontaneous activity fluctuations suffice to reconstruct the stimulus. As spontaneous activity is ubiquitous in the brain, modifiable fluctuations could provide an ideal substrate for Hebbian reverberations and sensory memory in other neural systems. PMID- 16354379 TI - The optimal synapse for sparse, binary signals in the rod pathway. AB - The sparsity of photons at very low light levels necessitates a nonlinear synaptic transfer function between the rod photoreceptors and the rod-bipolar cells. We examine different ways to characterize the performance of the pathway: the error rate, two variants of the mutual information, and the signal-to-noise ratio. Simulation of the pathway shows that these approaches yield substantially different performance at very low light levels and that maximizing the signal-to noise ratio yields the best performance when judged from simulated images. The results are compared to recent data. PMID- 16354380 TI - Simultaneous rate-synchrony codes in populations of spiking neurons. AB - Firing rates and synchronous firing are often simultaneously relevant signals, and they independently or cooperatively represent external sensory inputs, cognitive events, and environmental situations such as body position. However, how rates and synchrony comodulate and which aspects of inputs are effectively encoded, particularly in the presence of dynamical inputs, are unanswered questions. We examine theoretically how mixed information in dynamic mean input and noise input is represented by dynamic population firing rates and synchrony. In a subthreshold regime, amplitudes of spatially uncorrelated noise are encoded up to a fairly high input frequency, but this requires both rate and synchrony output channels. In a suprathreshold regime, means and common noise amplitudes can be simultaneously and separately encoded by rates and synchrony, respectively, but the input frequency for which this is possible has a lower limit. PMID- 16354381 TI - Spontaneous dynamics of asymmetric random recurrent spiking neural networks. AB - In this letter, we study the effect of a unique initial stimulation on random recurrent networks of leaky integrate-and-fire neurons. Indeed, given a stochastic connectivity, this so-called spontaneous mode exhibits various nontrivial dynamics. This study is based on a mathematical formalism that allows us to examine the variability of the afterward dynamics according to the parameters of the weight distribution. Under the independence hypothesis (e.g., in the case of very large networks), we are able to compute the average number of neurons that fire at a given time-the spiking activity. In accordance with numerical simulations, we prove that this spiking activity reaches a steady state. We characterize this steady state and explore the transients. PMID- 16354382 TI - Bayesian population decoding of motor cortical activity using a Kalman filter. AB - Effective neural motor prostheses require a method for decoding neural activity representing desired movement. In particular, the accurate reconstruction of a continuous motion signal is necessary for the control of devices such as computer cursors, robots, or a patient's own paralyzed limbs. For such applications, we developed a real-time system that uses Bayesian inference techniques to estimate hand motion from the firing rates of multiple neurons. In this study, we used recordings that were previously made in the arm area of primary motor cortex in awake behaving monkeys using a chronically implanted multielectrode microarray. Bayesian inference involves computing the posterior probability of the hand motion conditioned on a sequence of observed firing rates; this is formulated in terms of the product of a likelihood and a prior. The likelihood term models the probability of firing rates given a particular hand motion. We found that a linear gaussian model could be used to approximate this likelihood and could be readily learned from a small amount of training data. The prior term defines a probabilistic model of hand kinematics and was also taken to be a linear gaussian model. Decoding was performed using a Kalman filter, which gives an efficient recursive method for Bayesian inference when the likelihood and prior are linear and gaussian. In off-line experiments, the Kalman filter reconstructions of hand trajectory were more accurate than previously reported results. The resulting decoding algorithm provides a principled probabilistic model of motor-cortical coding, decodes hand motion in real time, provides an estimate of uncertainty, and is straightforward to implement. Additionally the formulation unifies and extends previous models of neural coding while providing insights into the motor cortical code. PMID- 16354383 TI - Facial attractiveness: beauty and the machine. AB - This work presents a novel study of the notion of facial attractiveness in a machine learning context. To this end, we collected human beauty ratings for data sets of facial images and used various techniques for learning the attractiveness of a face. The trained predictor achieves a significant correlation of 0.65 with the average human ratings. The results clearly show that facial beauty is a universal concept that a machine can learn. Analysis of the accuracy of the beauty prediction machine as a function of the size of the training data indicates that a machine producing human-like attractiveness rating could be obtained given a moderately larger data set. PMID- 16354384 TI - Classification of faces in man and machine. AB - We attempt to shed light on the algorithms humans use to classify images of human faces according to their gender. For this, a novel methodology combining human psychophysics and machine learning is introduced. We proceed as follows. First, we apply principal component analysis (PCA) on the pixel information of the face stimuli. We then obtain a data set composed of these PCA eigenvectors combined with the subjects' gender estimates of the corresponding stimuli. Second, we model the gender classification process on this data set using a separating hyperplane (SH) between both classes. This SH is computed using algorithms from machine learning: the support vector machine (SVM), the relevance vector machine, the prototype classifier, and the K-means classifier. The classification behavior of humans and machines is then analyzed in three steps. First, the classification errors of humans and machines are compared for the various classifiers, and we also assess how well machines can recreate the subjects' internal decision boundary by studying the training errors of the machines. Second, we study the correlations between the rank-order of the subjects' responses to each stimulus the gender estimate with its reaction time and confidence rating-and the rank order of the distance of these stimuli to the SH. Finally, we attempt to compare the metric of the representations used by humans and machines for classification by relating the subjects' gender estimate of each stimulus and the distance of this stimulus to the SH. While we show that the classification error alone is not a sufficient selection criterion between the different algorithms humans might use to classify face stimuli, the distance of these stimuli to the SH is shown to capture essentials of the internal decision space of humans. Furthermore, algorithms such as the prototype classifier using stimuli in the center of the classes are shown to be less adapted to model human classification behavior than algorithms such as the SVM based on stimuli close to the boundary between the classes. PMID- 16354388 TI - Introduction - The coxib controversies. AB - Much concern has arisen in the past year concerning the occurrence of serious cardiovascular adverse reactions from rofecoxib, celecoxib and some other members of the coxib group of NSAIDs. Factors underlying the sudden appearance of these events have been proposed to include the use of high doses of the drugs, undue reliance on their safety profile from controlled clinical trails and potent marketing leading to exposure of a large population who would inevitably present risks of cardiovascular adverse events. Suggestions are presented for future uses of the coxibs and approaches for their more cautious use and marketing. The use of the term "COX-2 inhibitor" to describe the coxibs is unhelpful and should be avoided since all NSAIDs have some degree of COX-2 inhibitory effect, though they may vary in their selectivity. Coxibs should be set aside in a group of their own within the broader category of the NSAIDs in view of both their cardiovascular risk and unique pharmacology. PMID- 16354389 TI - The use of NSAIDs in rheumatic disorders 2005: a global perspective. AB - Population studies and World Health Organisation (WHO) statistics indicate that 10-50% of individuals suffer from musculoskeletal disorders. Up to 3% will be classified as disabled due to their bone and joint condition, and the majority will suffer from pain. Almost all will require non-steroidal anti-inflammatory drugs (NSAIDs) and other analgesics for their management. The large majority of this population is elderly and, hence, at greater risk of adverse effects to the NSAIDs. The NSAIDs are a necessary choice in pain management because of the integrated role of the cyclo-oxygenase (COX) pathway in the generation of inflammation and in the biochemical recognition of pain. For over 80 years the management of musculoskeletal pain was hampered by NSAID toxicity problems related to the traditional NSAIDs. In the early 1990s, paracetamol was recommended as the first-choice analgesic for osteoarthritis, but subsequent studies have shown that paracetamol has a significant gastrointestinal (GI) toxicity profile. In addition, it has lower analgesic efficacy than NSAIDs and is, thus, not an effective alternative to NSAIDs in any of the inflammatory arthritides. The identification of cyclo-oxygenase 2 (COX-2) and the subsequent introduction of the COX-2-selective inhibitor NSAID drugs was thought to be a major breakthrough with the expectation of a significant reduction in G/I side effects. This has not been the case for celecoxib, and indeed for all COX-2 selective inhibitors when given with ASA. The COX-2-selective inhibitors also inhibit renal COX-2 with the potential for problems of fluid retention, oedema, hypertension and congestive heart failure. The major controversy with respect to the COX-2-selective inhibitors as a class has been the increase in myocardial infarction and other cardiovascular events observed in some studies. Thus, the initial expected global benefits of the COX-2-selective inhibitors may be outweighed by their potential for toxicity. Recent studies have shown that the use of a proton-pump inhibitor drug with traditional NSAIDs and with COX-2 selective inhibitors has been shown to significantly reduce GI symptoms and peptic ulceration. Thus, the traditional NSAIDs have been re-established as the preferred choice in the management of arthritis and musculoskeletal disorders. PMID- 16354390 TI - The murky waters of the coxibs: a review of the current state of play. AB - Since their development and licensing no class of drug has received as much attention both in the scientific and lay press as the cyclo-oxygenase 2 (COX-2) inhibitors. These compounds, also known as coxibs, were developed as a safer alternative to traditional non-steroidal anti-inflammatory drugs (NSAIDs) due to their reduced propensity to cause gastrointestinal (GI) irritation. However, an unforeseen complication was identified of increased cardiovascular (CV) morbidity and mortality. This observation threw into question the true 'safety' of the class and their subsequent role, if any, in patient management. The amount of information and misinformation regarding the coxibs is vast and as this field is in a state of flux much more will be forthcoming. This article will attempt to review the data regarding coxibs and make some recommendations regarding their ongoing use. PMID- 16354391 TI - The murky world of the COX-2-selective agents. PMID- 16354392 TI - COX-2 controversy: where are we and where do we go from here? AB - Much has been written in the last few months on the association between the cyclooxygenase-2 (COX-2)-selective inhibitors and cardiovascular events. We summarize the randomized controlled studies of different COX-2 inhibitors that led to rofecoxib withdrawal, provide evidence on the differences between various selective coxibs and summarize recommendations from the FDA and the American College of Rheumatology on use of COX-2 inhibitors. We give suggestions on the use of coxibs and briefly mention a general approach to the future of coxibs. PMID- 16354393 TI - Coxibs and their aftermath: an (opinionated) commentary, based on some historical viewpoints. AB - Five historical viewpoints are presented, each foreshadowing to some degree that it was only a matter of time before we would (re)learn that imbalanced anti prostanoid therapy was a potentially lethal strategy. Positive outcomes from this coxib debacle might be more willingness to (i) acknowledge our lack of understanding concerning normal homeostatic mechanisms involving the autopharmacology and pleiotropic functions of prostanoids and acyclic eicosanoids (leukotrienes, lipoxins, resolvins, etc.); (ii) revisit the question of salicylate versus aspirin for therapies not requiring anti-thrombotic prophylaxis; (iii) re-examine some leads provided by natural/alternate therapies for inflammatory disorders; and (iv) reconsider the downside of over-reliance upon the present patent system as the main shield and driver of drug innovation. PMID- 16354394 TI - COX-2 inhibitors: a story of greed, deception and death. AB - In 1999, drug manufacturers introduced a class of NSAIDs called COX-2 inhibitors or coxibs. The drugs were avidly promoted directly to the consumers and became bestsellers from the start. Arthritis sufferers were eager to take medications that eased joint pain with less risk of causing gastrointestinal pain, bleeding and other side-effects. In the year after their introduction, doctors wrote over 100 million prescriptions for celecoxib (Celebrex) and rofecoxib (Vioxx). Celebrex is the sixth best-selling drug, with sales of more than US$ 4 billion since its debut in 1999. Vioxx had sales of US$ 2.6 billion in 2001. However, the coxibs increase the risk of heart attacks and strokes, and their price, in the USA, is obscene. The manufacturers faced a possibly complicit, toothless and bloodless FDA, and used every maneuvering to fleece the patients. We must now reflect on attitudes that we thought only belong to the tobacco industry. Fortunately, safe and active alternatives exist. PMID- 16354395 TI - Pharmacological actions of Cordyceps, a prized folk medicine. AB - Cordyceps species, including C. sinensis, C. militaris, C. pruinosa and C. ophioglossoides, are prized traditional medicinal materials. The aim of this article is to review the chemical constituents and pharmacological actions of Cordyceps species. The chemical constituents include cordycepin (3'-de oxyadenosine) and its derivatives, ergosterol, polysaccharides, a glycoprotein and peptides containing alpha-aminoisobutyric acid. They include anti-tumour, anti-metastatic, immunomodulatory, antioxidant, anti-inflammatory, insecticidal, antimicrobial, hypolipidaemic, hypoglycaemic, anti-ageing, neuroprotective and renoprotective effects. Polysaccharide accounts for the anti-inflammatory, antioxidant, anti-tumour, anti-metastatic, immunomodulatory, hypoglycaemic, steroidogenic and hypolipidaemic effects. Cordycepin contributes to the anti tumour, insecticidal and antibacterial activity. Ergosterol exhibits anti-tumour and immunomodulatory activity. A DNase has been characterized. PMID- 16354396 TI - Controlled release tamsulosin hydrochloride from alginate beads with waxy materials. AB - The objective of this study was to develop oral controlled release delivery systems for tamsulosin hydrochloride (TSH) using alginate beads with various waxy materials, such as Compritol 888 ATO, Precirol ATO 5 and Gelucires. The beads were prepared from sodium alginate-waxy material-TSH slurry dropped onto calcium chloride to form spherical beads. The effects of the addition of various waxy materials to alginate beads on the drug encapsulation efficiency, bead size and morphology were investigated. The drug encapsulation efficiency significantly increased with the addition of waxy materials. The TSH-loaded alginate beads with and without waxy materials were almost spherical particles with an average diameter of 1.44 and 1.22 mm, respectively. In dissolution study, the TSH-loaded alginate beads with waxy materials exhibited controlled release behaviour over a 6-h period, while beads without waxy materials showed release of 100% TSH within 2 h. These results may be attributed to the formation of a more rigid alginate matrix structure due to incorporated waxy materials. From the Dunnett's t-test and the f2 factor, the release of TSH from alginate beads, a similar dissolution pattern to that of the marketed product (Harunal capsules) could be achieved by adding Gelucire 50/13 into TSH-loaded alginate beads. From these results, oral controlled release of TSH could be achieved with loading in alginate beads with waxy materials, such as Compritol 888 ATO, Precirol ATO 5 and Gelucires. PMID- 16354397 TI - Cefuroxime axetil solid dispersions prepared using solution enhanced dispersion by supercritical fluids. AB - Cefuroxime axetil (CA) solid dispersions with HPMC 2910/PVP K-30 were prepared using solution enhanced dispersion by supercritical fluids (SEDS) in an effort to increase the dissolution rate of poorly water-soluble drugs. Their physicochemical properties in solid state were characterized by differential scanning calorimeter (DSC), powder X-ray diffraction (PXRD), Fourier transform infrared spectrometry (FT-IR) and scanning electron microscopy. No endothermic and characteristic diffraction peaks corresponding to CA were observed for the solid dispersions in DSC and PXRD. FTIR analysis demonstrated the presence of intermolecular hydrogen bonds between CA and HPMC 2910/PVP K-30 in solid dispersions, resulting in the formation of amorphous or non-crystalline CA. Dissolution studies indicated that the dissolution rates were remarkably increased in solid dispersions compared with those in the physical mixture and drug alone. In conclusion, an amorphous or non-crystalline CA solid dispersion prepared using SEDS could be very useful for the formulation of solid dosage forms. PMID- 16354398 TI - Clinical pharmacokinetics of the cyclooxygenase inhibiting nitric oxide donator (CINOD) AZD3582. AB - The clinical pharmacokinetics of the COX-inhibiting nitric oxide donator (CINOD) AZD3582 and its metabolites, including naproxen, nitric oxide and nitrate, are summarized. AZD3582 has low aqueous solubility, moderate and passive intestinal permeability and is degraded by intestinal esterases. Its oral bioavailability (F) appears to be maximally a few per cent, and increases by several-fold after food intake. Ninety-four per cent or more of an AZD3582 dose is absorbed, of which at least 9-20% appears to be taken up as intact substance. AZD3582 has a predicted plasma protein binding degree of approximately 0.1%, a half-life (t1/2) of 3 to 10 h and does not accumulate after repeated once- and twice-daily dosing. In patients AZD3582 does not provide a significantly better gastrointestinal (GI) side-effect profile than the highly permeable and locally irritating naproxen. Possible reasons for this include considerable GI uptake as naproxen, limited duration and extent of nitric oxide donation in the GI mucosa and the circulation, tolerance development (involving auto-inhibition of nitric oxide catalysing enzymes) and mucosal damage caused by nitric oxide. Blood pressure data suggest that nitric oxide is mainly donated within 3 h. The uptake of naproxen is slightly slower and lower (> or = 94% relative GI uptake and 80-85% relative F) after AZD3582 administration compared with naproxen dosing. The naproxen t1/2 and trough steady-state concentrations after AZD3582 and naproxen dosing are similar. The average systemic nitrate exposure is approximately doubled after dosing of 375 to 750 mg AZD3582 twice daily. PMID- 16354399 TI - Size-dependent disposition of nanoparticles and microparticles following subconjunctival administration. AB - The purpose of this study was to determine the retention and ocular distribution of subconjunctivally administered nanoparticles and microparticles. Fluorescent polystyrene particles (carboxylate modified, negatively charged) of various sizes (20 nm, 200 nm and 2 microm; Fluospheres, dose 400 microg) were administered to male Sprague-Dawley rats by subconjunctival injection under anaesthesia. The disposition of the particles in the periocular and ocular tissues was studied for up to 60 days by quantifying the particle amounts using liquid extraction followed by spectrofluorimetric analysis. The effect of dose on the particle disposition was investigated with a 40-microg dose of the particles. The effect of an increase in surface hydrophobicity was evaluated for the 20 and 200 nm particles at 1 day post administration. Following periocular administration, penetration into the ocular tissues was negligible for the carboxylate-modified microparticles as well as nanoparticles. Almost the entire dose of the 200 nm and 2 microm particles was retained in the periocular tissue at 60 days post administration. The 20 nm particles disappeared rapidly from the periocular tissue with 15 and 8% of administered dose remaining after 1 and 7 days, respectively. The 20 nm particles could not be detected in the periocular tissue at 60-days post-administration. An increase in the surface hydrophobicity did not affect the periocular retention of 200 nm particles but elevated that of the 20 nm particles, at the end of day 1. It was concluded that subconjunctivally administered 200 nm and larger particles can be almost completely retained at the site of administration for at least two months. Periocular administration of particulate systems of this size would likely be useful as sustained drug delivery systems. PMID- 16354400 TI - Sulfasalazine transport in in-vitro, ex-vivo and in-vivo absorption models: contribution of efflux carriers and their modulation by co-administration of synthetic nature-identical fruit extracts. AB - Sulfasalazine is characterised by low oral bioavailability. In this study, its intestinal transport characteristics were studied in an in-vitro, ex-vivo and in situ system. The absorptive transport of sulfasalazine across Caco-2 monolayers appeared to be lower than the secretory transport (P(app-abs) = 0.21 +/- 0.02 x 10(-6) cm s(-1) and P(app-secr) = 2.97 +/- 0.30 x 10(-6) cm s(-1), respectively). This polarity in transport of sulfasalazine was not mediated by P-glycoprotein (P gp), as inclusion of verapamil (100 microM) did not have any effect on the transport polarity of sulfasalazine. However, inclusion of the multidrug resistance-associated protein (MRP) inhibitors benzbromarone (50 microM) and sulfinpyrazone (1 mM), and the glutathione-depleting agent chlorodinitrobenzene (100 microM), resulted in an increased absorptive transport of sulfasalazine in the Caco-2 system (P(app-abs) = 0.64 +/- 0.02, 0.51 +/- 0.04 and 0.60 +/- 0.03 x 10(-6) cm s(-1), respectively). The interference of carriers implies that, during absorption, interactions with food components may occur at the level of this carrier. Therefore, the effect of food extracts was studied in a parallel set of experiments. For two standardized nature-identical fruit extracts (pineapple and apricot extract) a concentration-dependent absorption-enhancing effect could be observed in the Caco-2 system. The functional expression of similar carriers was also demonstrated in rat ileum in the Ussing chamber system. Interaction studies with fruit extracts in the Ussing chamber system, as well as in the in-situ intestinal perfusion study, revealed a 2- to 4-fold increase in the absorptive transport of sulfasalazine. These results indicate that food components in the intestinal lumen can have a significant impact on the intestinal absorption characteristics of sulfasalazine by modulating the biochemical barrier function of the intestinal mucosa. PMID- 16354401 TI - Antidiabetic effect of a new peptide from Squalus mitsukurii liver (S-8300) in streptozocin-induced diabetic mice. AB - We have evaluated the antidiabetic effect of S-8300 (a peptide extracted from shark liver (Squalus mitsukurii)) in streptozocin (streptozotocin)-diabetic mice. Diabetes was induced by a single intravenous injection of streptozocin (150 mg kg(-1)). The effects of S-8300 (3 or 10 mg kg(-1)) on diabetic mice were investigated by observing the changes in the levels of fasting plasma glucose, glycosylated haemoglobin, hepatic glycogen, triglycerides, cholesterol, free fatty acid, superoxide dismutase, and malondialdehyde. Body weight, kidney weight and the degree of injured beta-cells in pancreatic islets were recorded also. Diabetic mice treated with S-8300 showed a significant decrease in the levels of fasting plasma glucose, glycosylated haemoglobin, triglycerides, cholesterol, free fatty acid in plasma and malondialdehyde in tissues. The animals showed a significant increase in the content of hepatic glycogen and the activity of superoxide dismutase. Treatment with S-8300 attenuated the degree of injured beta cells in the pancreatic islets. The effect of S-8300 was similar to that of glibenclamide (5 mg kg(-1)). PMID- 16354402 TI - Cytoprotective mechanism of baicalin against endothelial cell damage by peroxynitrite. AB - To evaluate the mechanism responsible for the cytoprotective effects of baicalin, an antioxidant flavonoid isolated from Scutellaria baicalensis, we investigated its effects against peroxynitrite (ONOO-)-induced endothelial cell (EC) damage. Baicalin showed efficient antioxidative actions by its ability to scavenge ONOO and inhibit ONOO(-)-mediated nitrotyrosine formation in vitro. Using an EC (YPEN 1) culture system, baicalin exhibited cytoprotective effects against cell death by ONOO- that was induced exogenously with tert-butyl hydroperoxide (t-BHP) in the YPEN-1 model. Baicalin was also found to reduce the intracellular precursors of ONOO-, NO* and O2(-) in the t-BHP-treated ECs. Evidence from Western blotting further revealed down-regulated expressions of iNOS and COX2, endogenous sources of NO* and O2(-) by baicalin treatment. In addition, pre-incubation of baicalin with EC suppressed t-BHP-induced nuclear factor kappa-B binding activity as determined by the transfection assay and Western blot analysis, further indicating baicalin's inhibition of iNOS and COX-2 expression. Based on the present data, we propose that baicalin scavenges ONOO- and protects cells against injury. Based on these data, it was concluded that baicalin is potentially a useful antioxidant against ONOO- and NO* and an inhibitor of iNOS and COX-2. PMID- 16354403 TI - Apoptotic potential of sesquiterpene lactone ergolide through the inhibition of NF-kappaB signaling pathway. AB - Treatment with ergolide, a sesquiterpene lactone from Inula britannica var chinensis, caused the induction of apoptosis in Jurkat T cells, which was confirmed by DNA fragmentation, caspase-3 activation and cleavage of poly(ADP ribose) polymerase in response to ergolide. Furthermore, mitochondrial dysfunction appeared to be associated with ergolide-induced apoptosis, because Bax translocation and cytochrome c release were stimulated by ergolide. In parallel, the nuclear factor-kappaB (NF-kappaB) signaling pathway was significantly inhibited by ergolide, which was accompanied by down-regulation of cell survival molecules, such as X-chromosome-linked inhibitor of apoptosis and Bcl-2. In addition, the JNK signaling pathway was involved in ergolide-induced apoptosis. Collectively, our results identified a new mechanism for the anti cancer property of ergolide, attributable to the induction of apoptosis through down-regulation of cell survival signal molecules resulting from inhibition of the NF-kappaB signaling pathway. PMID- 16354404 TI - The role of endothelin receptor antagonism in collar-induced intimal thickening and vascular reactivity changes in rabbits. AB - Intimal thickening, due to smooth muscle cell migration and proliferation, is considered to be one of the major components of vascular proliferative disorders such as atherosclerosis and restenosis. One experimental model, resulting in intimal thickening in the rabbit, involves placing a silicon collar around the carotid artery, and is used in this study. Endothelin is known to act as a strong mitogen and to stimulate smooth muscle cell proliferation and migration. We investigated the contribution of endothelin to the development of collar-induced intimal thickening and the effects of TAK-044, (5 mg kg(-1) daily, s.c.), a non selective ET(A)/ET(B) receptor antagonist, on intimal thickening and vascular reactivity changes in the collared rabbit carotid artery. Endothelin levels and the intimal cross-sectional area, as well as the ratio of intimal area to media (index), increased significantly in collared arteries as compared with those in sham-operated arteries. TAK-044 significantly inhibited intimal thickening and also decreased the index without affecting increased endothelin levels in collared arteries. Vascular reactivity changes in response to collaring produced predictable effects, such as decreased contractile responses to vasoconstrictor agents and increased sensitivity to serotonin (5-hydroxytryptamine, 5-HT). In terms of contractile responses in this model, TAK-044, in particular, did not affect collar-induced vascular reactivity changes. These results suggest that endothelin may be involved in the pathogenesis of collar-induced intimal thickening. As an endothelin receptor antagonist, TAK-044 may potentially be beneficial in the treatment of atherosclerosis. PMID- 16354405 TI - The phosphodiesterase 4 inhibitor AWD 12-281 is active in a new guinea-pig model of allergic skin inflammation predictive of human skin penetration and suppresses both Th1 and Th2 cytokines in mice. AB - The selective phosphodiesterase 4 (PDE4) inhibitor AWD 12-281 is structurally optimized for topical administration. It has potent effects in models of lung inflammation if administered as a dry powder inhalation. It has also demonstrated its anti-inflammatory property in a mouse model of cutaneous inflammation after topical administration. The aim of this study was to evaluate whether AWD 12-281 may be capable of penetrating human skin. Therefore a new guinea-pig model of allergic skin inflammation had to be developed. In ovalbumin-sensitized guinea pigs, intracutaneous administration of ovalbumin results in a rapid development of allergic skin wheals. Topically administered AWD 12-281 was capable of reducing the development of wheals, indicating that this compound can penetrate the stratum corneum of guinea-pig skin as a predictor of human skin penetration. A secondary aim was the evaluation of a T cell subtype preference of AWD 12-281 since PDE4 inhibitors are said to preferentially inhibit Th2-type cytokines. Therefore, the effects of AWD 12-281 on a broad spectrum of Th1- and Th2-type cytokines were studied in tissue homogenates after allergen challenge in sensitized mice and in supernatants of anti CD3/anti-CD28-stimulated peripheral blood mononuclear cells (PBMCs). In both models, AWD 12-281 suppressed both T cell subtype cytokines indicating a broad spectrum activity of AWD 12-281. A further issue was to determine the duration of action and the concentration response relationship of the topical activity of AWD 12-281 using a model of acute local inflammation--the arachidonic-acid-induced mouse ear oedema. The compound exhibited a dose-dependent effect with a minimally effective concentration of 0.3%; after repeated administration the minimally effective concentration was found to be 0.03%. A single administration of a 3% solution resulted in significant suppression of inflammation even 48 h after treatment. In conclusion, our results indicate that AWD 12-281 is a very promising drug candidate not only for the treatment of lung inflammation using inhalative administration but also for the treatment of atopic dermatitis. PMID- 16354406 TI - Preparation and stereochemical integrity of certain thioesters of 2-arylpropionic acids and related compounds. AB - 2-Arylpropionate thioesters 5, 6a/6b and 7a/7b, 2-aryloxypropionate thioesters 8a/8b and 2-alkoxy-2-arylacetate thioesters 9a/9b were prepared from thiol 4 and the corresponding carboxylic acids. Thioesters 6a/6b, 7a/7b, 8a/8b and 9a/9b were monitored for evidence of inter-conversion between epimers in acetonitrile solvent at 40 degrees C, by optical activity in the cases of 6a/6b and 7a/7b, and by 1H NMR spectroscopy in the cases of 8a/8b and 9a/9b. Only in the case of thioesters 9a/9b was significant inter-conversion between epimers observed. PMID- 16354407 TI - Gastroprotection and effect of the simultaneous administration of Cuachalalate (Amphipterygium adstringens) on the pharmacokinetics and anti-inflammatory activity of diclofenac in rats. AB - This work aimed to study the effect of Cuachalalate methanol extract (CME) on the anti-inflammatory activity and pharmacokinetics of diclofenac sodium, a frequently prescribed non-steroidal anti-inflammatory drug (NSAID). The gastroprotective effect of CME on the gastric injury induced by diclofenac was studied in rats. CME showed a gastroprotective effect of 15.7% at 1 mg kg(-1) and 72.5% at dose of 300 mg kg(-1). Omeprazole, used as anti-ulcer reference drug, showed gastroprotective effects of 50-89.7% at doses tested (1-30 mg kg(-1)). The value of the 50% effective dose for the anti-inflammatory effect of diclofenac sodium (ED50 = 1.14 +/- 0.23 mg kg(-1)) using carrageenan-induced rat paw oedema model, was not modified by the concomitant administration of 30 or 100 mg kg(-1) of CME. The effect of CME (30, 100 and 300 mg kg(-1), p.o.) on the pharmacokinetics of diclofenac sodium was studied. It was observed that the simultaneous administration of diclofenac sodium and 300 mg kg(-1) of CME decreased significantly the values of Cmax (7.08 +/- 1.42 microg mL(-1)) and AUC (12.67 +/- 2.97 microg h mL(-1)), but not the value of tmax (0.13 (0.1-0.25)h) obtained with the administration of diclofenac alone. The simultaneous administration of 30 or 100 mg kg(-1) of CME did not modify the pharmacokinetic parameters of diclofenac. The experimental findings in rats suggest that CME at doses lower than 100 mg kg(-1) protects the gastric mucosa from the damage induced by diclofenac sodium without altering either the anti-inflammatory activity or the pharmacokinetics of this NSAID. PMID- 16354408 TI - Protective effect of Hachimi-jio-gan against renal failure in a subtotal nephrectomy rat model. AB - The protective effect of Hachimi-jio-gan extract against chronic renal failure in a subtotal nephrectomy rat model was investigated. The level of serum urea nitrogen by nephrectomy was increased over 15 weeks, but the administration of Hachimi-jio-gan at 50 and 200 mg led to the decrease. In addition, the levels of creatinine (Cr), urinary methylguanidine (MG) and MG/Cr were increased, whereas Cr clearance dramatically decreased in nephrectomized rats. However, oral administration of Hachimi-jio-gan extract prevented the elevation of these uremic toxins in serum and urine, and the production of hydroxyl radical. Moreover, nephrectomy led to a significant decline in superoxide dismutase (SOD) and catalase activities, but increased glutathione peroxidase activity compared with normal levels, indicating an abnormal antioxidative system. The increased activity of both SOD and catalase by the oral administration of Hachimi-jio-gan suggested that these enzymes are associated with the protective role of Hachimi jio-gan extract against oxidative stress by nephrectomy. Moreover, the decrease in serum albumin in nephrectomized control rats was increased and proteinuria was ameliorated by the administration of Hachimi-jio-gan with improved glomerular hyalinosis, interstitial fibrosis and inflammation, suggesting the beneficial effect of Hachimi-jio-gan to prevent glomerular sclerosis and progressive renal fibrosis. This study suggests that Hachimi-jio-gan plays a protective role in the progression of chronic renal failure through the decline in uremic toxins, elevation of antioxidative enzyme activity such as SOD and catalase, and amelioration of histopathological lesions in the kidney. PMID- 16354409 TI - Quantitative analysis of the major constituents of St John's wort with HPLC-ESI MS. AB - A method was developed to profile the major constituents of St John's wort extracts using high-performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI-MS). The objective was to simultaneously separate, identify and quantify hyperforin, hypericin, pseudohypericin, rutin, hyperoside, isoquercetrin, quercitrin and chlorogenic acid using HPLC-MS. Quantification was performed using an external standardisation method with reference standards. The method consisted of two protocols: one for the analysis of flavonoids and glycosides and the other for the analysis of the more lipophilic hypericins and hyperforin. Both protocols used a reverse phase Luna phenyl hexyl column. The separation of the flavonoids and glycosides was achieved within 35 min and that of the hypericins and hyperforin within 9 min. The linear response range in ESI MS was established for each compound and all had linear regression coefficient values greater than 0.97. Both protocols proved to be very specific for the constituents analysed. MS analysis showed no other signals within the analyte peaks. The method was robust and applicable to alcoholic tinctures, tablet/capsule extracts in various solvents and herb extracts. The method was applied to evaluate the phytopharmaceutical quality of St John's wort preparations available in the UK in order to test the method and investigate if they contain at least the main constituents and at what concentrations. PMID- 16354410 TI - Methylisogermabullone isolated from radish roots stimulates small bowel motility via activation of acetylcholinergic receptors. AB - We have previously reported that extract of radish roots exhibits an increase in gastrointestinal motility through the activation of muscarinic acetylcholine (ACh) receptors. Based on the stimulatory activity-guided fractionation on rat ileal segments, this study isolated methylisogermabullone (MIGB, C23H31O5NS, MW 433) from methanol extracts of radish roots. MIGB caused a significant increase of the isolated rat ileal contraction in a concentration-dependent manner (23-693 microM), and the pattern of MIGB-induced ileal contraction was different in the time course to that produced by ACh. The EC50 value of MIGB, to produce 50% maximum ileal contraction, was estimated to be 45.5 microM. MIGB (230 microM) induced ileal contractions were enhanced by pretreatment of segments with ACh (0.1 microM). Ileal contractions produced by MIGB (230 microM) or ACh (0.1 microM) at submaximal concentration were partially inhibited by pretreatment of hexamethonium (0.1 mM), a ganglionic blocker, whereas they were almost completely abolished by atropine (10 microM). Oral administration of MIGB to mice stimulated the small intestinal transit of charcoal in a dose-dependent manner (10-100 mg kg(-1)), and MIGB (100 mg kg(-1))-induced stimulation of small intestinal transit was significantly attenuated by co-administration of atropine (50 mg kg(-1)). Taken together, these results demonstrate that MIGB isolated from radish roots stimulates the small bowel motility through the activation of ACh receptors. These findings suggest that MIGB may become a potential regulatory agent for therapeutic intervention in dysfunction of gastrointestinal motility. PMID- 16354411 TI - Inhibition by licochalcone A, a novel flavonoid isolated from liquorice root, of IL-1beta-induced PGE2 production in human skin fibroblasts. AB - Licochalcone A, a novel flavonoid isolated from the root of Glycyrrhiza inflata, has been reported to exhibit anti-inflammatory activity in animal models. In this study, we examined the effect of licochalcone A on the production of chemical mediators such as prostaglandin (PG)E2 and cytokines by interleukin (IL)-1beta in human skin fibroblasts. Licochalcone A (IC50 15.0 nM) inhibited PGE2 production, but not IL-6 and IL-8 production, in response to IL-1beta. NS-398 (IC50 1.6 nM), a COX-2 selective inhibitor, also suppressed the PGE2 production. Furthermore, licochalcone A and NS-398 suppressed PGF(2alpha) production by IL-1beta. However, licochalcone A (1 microM) had no effect on increased levels of cyclooxygenase (COX)-2 mRNA and protein in cells. Dexamethasone (100 nM) not only inhibited PGE2, PGF(2alpha), IL-6 and IL-8 production but also strongly suppressed the expression of COX-2 mRNA and protein. Licochalcone A had no effect on COX-1 dependent PGE2 production, whereas indometacin (100 nM), a dual inhibitor of COX 1 and COX-2, was very effective. These results suggest that licochalcone A induces an anti-inflammatory effect through the inhibition of COX-2-dependent PGE2 production. Furthermore, it appears that the inhibitory effect of licochalcone A on PGE2 production in response to IL-1beta is quite different from that of the steroid. PMID- 16354412 TI - Direct oxidative modifications of signalling proteins in mammalian cells and their effects on apoptosis. AB - The production of ROS is an inevitable consequence of metabolism. However, high levels of ROS within a cell can be lethal and so the cell has a number of defences against oxidative cell stress. Occasionally the cell's antioxidant mechanisms fail and oxidative stress occurs. High levels of ROS within a cell have a number of direct and indirect consequences on cell signalling pathways and may result in apoptosis or necrosis. Although some of the indirect effects of ROS are well known, limitations in technology mean that the direct effects of the cell's redox environment upon proteins are less understood. Recent work by a number of groups has demonstrated that ROS can directly modify signalling proteins through different modifications, for example by nitrosylation, carbonylation, di-sulphide bond formation and glutathionylation. These modifications modulate a protein's activity and several recent papers have demonstrated their importance in cell signalling events, especially those involved in cell death/survival. Redox modification of proteins allows for further regulation of cell signalling pathways in response to the cellular environment. Understanding them may be critical for us to modulate cell pathways for our own means, such as in cytotoxic drug treatments of cancer cells. Protein modifications mediated by oxidative stress can modulate apoptosis, either through specific protein modifications resulting in regulation of signalling pathways, or through a general increase in oxidised proteins resulting in reduced cellular function. This review discusses direct oxidative protein modifications and their effects on apoptosis. PMID- 16354413 TI - Oxidative stress and antioxidant deficiencies in asthma: potential modification by diet. AB - The lungs of asthmatic patients are exposed to oxidative stress due to the generation of reactive oxygen and nitrogen species as a consequence of chronic airway inflammation. Increased concentrations of NO*, H2O2 and 8-isoprostane have been measured in exhaled breath and induced sputum of asthmatic patients. O2*-, NO*, and halides interact to form highly reactive species such as peroxynitrite and HOBr, which in turn cause nitration and bromination of protein tyrosine residues. Oxidative stress may also reduce glutathione levels and cause inactivation of antioxidant enzymes such as superoxide dismutase, with a consequent increase in apoptosis, shedding of airway epithelial cells and airway remodelling. The oxidant/antioxidant equilibrium in asthmatic patients may be further perturbed by low dietary intakes of the antioxidant vitamins C and E, selenium and flavonoids, with a consequent lowering of the concentrations of these and other non-dietary antioxidants such as bilirubin and albumin in plasma and airway epithelial lining fluid. Although supplementation with vitamins C and E appears to offer protection against the adverse effects of ozone, recent randomised, placebo-controlled trials of vitamin C or E supplements for patients with mild asthma have not shown significant benefits over standard therapy. However, genetic variation in glutathione S-transferase may influence the susceptibility of asthmatic individuals to oxidative stress and the extent to which they are likely to benefit from antioxidant supplementation. Long-term prospective trials are required to determine whether modification of dietary intake will benefit asthma patients and reduce the socio-economic burden of asthma in the community. PMID- 16354414 TI - Antioxidant and radioprotective properties of an Ocimum sanctum polysaccharide. AB - The antioxidant activity of two polysaccharides isolated from the Indian medicinal plants, Ocimum sanctum and Tinospora malabarica, was studied. Only the O. sanctum polysaccharide (OSP) showed significant activity. OSP could prevent oxidative damage to liposomal lipids and plasmid DNA induced by various oxidants such as iron, AAPH and gamma-radiation, besides scavenging important ROS such as the superoxide radical and hydrogen peroxide and inhibiting xanthine oxidase. In addition, OSP could prevent gamma-radiation-mediated cell deaths in mouse splenocytes. PMID- 16354415 TI - Biotransformation of nitric oxide in the cerebrospinal fluid of amyotrophic lateral sclerosis patients. AB - Recent findings indicate that nitric oxide (NO*) over-production might be an important factor in the pathogenesis of sporadic amyotrophic lateral sclerosis (SALS). We measured significantly higher concentrations of uric acid and thiol group-containing molecules (R-SH groups) in the cerebrospinal fluid (CSF) from SALS patients compared to controls. The above factors, together with a slightly increased free iron concentration found in the CSF, favour conditions necessary for the formation of the dinitrosyl iron complex, capable of NO* bio transformation. Thus, we performed ex vivo saturation of CSF (from both SALS patients and controls) with NO*. A decrease in the level of R-SH was found. This was more pronounced in the CSF from SALS patients. In the CSF from SALS patients the production of nitrite and hydroxylamine was greater than that observed in the CSF from controls. Moreover, we also found increased Cu,Zn-SOD activity in the CSF from SALS patients (when compared to control subjects) but no activity corresponding to Mn-SOD in any CSF samples. As Cu,Zn-SOD can react with nitroxyl forming NO*, the conditions for a closed, but continuous, loop of NO* biotransformation are present in the CSF of ALS patients. PMID- 16354417 TI - Mental health: a public health priority in the Americas. PMID- 16354419 TI - [Mental disorders in Latin America and the Caribbean: a public health priority]. AB - OBJECTIVE: The growing burden of mental disorders in Latin America and the Caribbean has become too large to ignore. There is a need to know more about the prevalence of mental disorders and the gap between the number of individuals with psychiatric disorders and the number of those persons who remain untreated even though effective treatments exist. Having that knowledge would make it possible to improve advocacy, adopt better policies, formulate innovative intervention programs, and apportion resources commensurate with needs. METHODS: Data were extracted from community-based psychiatric epidemiological studies published in Latin America and the Caribbean from 1980 through 2004 that used structured diagnostic instruments and provided prevalence rates. Estimates of the crude rates in Latin America and the Caribbean for the various disorders were determined by calculating the mean and median rates across the studies, by gender. In addition, data on service utilization were reviewed in order to calculate the treatment gap for specific disorders. RESULTS: Nonaffective psychosis (including schizophrenia) had an estimated mean one-year prevalence rate of 1.0%; major depression, 4.9%; and alcohol use abuse or dependence, 5.7%. Over one-third of individuals with nonaffective psychosis, over half of those with an anxiety disorder, and some three-fourths of those with alcohol use abuse or dependence did not receive mental health care from either specialized or general health services. CONCLUSIONS: The current treatment gap in mental health care in Latin America and the Caribbean remains wide. Further, current data likely greatly underestimate the number of untreated individuals. The epidemiological transition and changes in the population structure will further widen the treatment gap in Latin America and the Caribbean unless mental health policies are formulated or updated and programs and services are expanded. PMID- 16354420 TI - Alcohol consumption and burden of disease in the Americas: implications for alcohol policy. AB - OBJECTIVES: To describe patterns of alcohol consumption in the Americas, to estimate the burden of disease attributable to alcohol in the year 2000, and to suggest implications for policies to reduce alcohol-related disease burden. METHODS: Two dimensions of alcohol exposure were included in this secondary data analysis: average volume of alcohol consumption and patterns of drinking. There were two main outcome measures: mortality (number of deaths) and disability adjusted life years (DALYs) lost (number of years of life lost due to premature mortality and disability). Separate estimates were obtained for different sexes, age groups and WHO regions. RESULTS: Despite regional variations, alcohol consumption in the Americas averaged more than 50% higher than worldwide consumption. Patterns of irregular heavy drinking prevailed. Alcohol consumption caused a considerable disease burden: 4.8% of all the deaths and 9.7% of all DALYs lost in the year 2000 were attributable to drinking, with most of the burden occurring outside North America. Intentional and unintentional injuries accounted for 59.8% of all alcohol-related deaths and 38.4% of the alcohol related disease burden. Of all risk factors compared here, alcohol accounted for the greatest proportion of risk, followed by smoking. CONCLUSIONS: Interventions should be implemented to reduce the high burden of alcohol-related disease in the Americas. Given the epidemiological structure of this burden, injury prevention including, but not restricted to, prevention of traffic injuries, as well as appropriate treatment options, should play an important role in comprehensive plans to reduce the alcohol-related public health burden. PMID- 16354421 TI - [Substance use, risk situations, and HIV seroprevalence among individuals seeking free HIV testing in Porto Alegre, Brazil]. AB - OBJECTIVE: To describe substance use, HIV seroprevalence, and risk of exposure to HIV infection in individuals seeking HIV testing in two screening centers (a municipal one and a state one) in the city of Porto Alegre, Rio Grande do Sul, Brazil. METHODS: Using a cross-sectional design, we enrolled a convenience sample of 1 026 men and women between 15 and 60 years of age. We included all the subjects who, after an initial triage, reported any drug use or any behavior or situation with risk of HIV transmission. A Brazilian-Portuguese version of the Risk Assessment Battery was used to assess exposure to risk situations. Blood samples were tested for anti-HIV antibodies, using enzyme-linked immunosorbent assay. Before the blood test, individuals participated in a group counseling session (with a maximum of 20 participants) on HIV and AIDS, in line with the criteria of the Ministry of Health of Brazil. RESULTS: Among the sample, a high overall HIV seroprevalence was found, 15.1%. Drug use did not explain all of the risk exposure. In comparison to those who reported no injecting drug use, the odds ratio (OR) for being HIV-positive for those who reported injecting drug use at some point in their life was 7.6 (95% confidence interval (CI) = 4.4 to 13.0). However, only 10.3% of the sample reported any injecting drug use. The variables that were associated with HIV seropositivity were: male gender (OR = 1.8; 95% CI = 1.1 to 2.8), monthly family income below three times the minimum-salary amount (OR = 2.1; 95% CI = 1.3 to 3.5), age over 25 (OR = 1.7; 95% CI = 1.1 to 2.7), and having had sexual relations with a partner who was possibly HIV-positive (OR = 1.8; 95% CI = 1.1 to 3.2). CONCLUSIONS: Even irregular or occasional drug use increases the odds of seropositivity, particularly if intravenous drugs are used. Sexual transmission played an important role in seropositivity in this sample, indicating that drug use can affect the judgment of risk and thus contribute to HIV transmission. PMID- 16354422 TI - The epidemiology of schizophrenia and other common mental health disorders in the English-speaking Caribbean. AB - OBJECTIVE: To report on the epidemiology of schizophrenia and other common mental health disorders in the English-speaking Caribbean. METHODS: The MEDLINE, MEDCARIB and West Indiana electronic databases were searched with the phrases Caribbean AND psychiatry OR mental OR suicide, and items with specific reference to epidemiology were culled and reviewed. RESULTS: The age-corrected incidence rate for schizophrenia per 10,000 is 2.09 in Jamaica, 2.2 per 1000 in Trinidad, and 2.92 in Barbados. These rates are lower than the incidence rates reported for white British people, and significantly lower that the 6- to 18-fold higher risk ratio incidence reported for African Caribbeans living in Britain. A comparative diagnostic study carried out in the United Kingdom (UK) suggests that misdiagnosis plays a significant role in this difference. Relatively low incidence figures for affective disorders, anxiety states, suicide and attempted suicide have been reported for Jamaica, Trinidad, and Barbados. DISCUSSION: Most published epidemiological studies of Caribbean populations center on schizophrenia, because of the startling findings of a large increase in risk ratios for African Caribbean people living in England compared to the white native British people. The etiological evidence is shifting toward factors of racism and social alienation experienced by black people in the UK, and to misdiagnosis by white British psychiatrists. Studies of the role of colonial enslavement and social engineering raise the etiological possibility of a role for social and structural community organization in the genesis of schizophrenia. The Caribbean thus represents a rich crucible for research in the epidemiology and etiology of mental health disorders. PMID- 16354423 TI - [Service use patterns among adults with mental health problems in Chile]. AB - OBJECTIVE: To describe the patterns in the use of general health services and specialized health services among adults with mental health problems in Chile, as well as those persons' level of satisfaction with the services. The overall objective was to optimize the use of the limited resources available for mental health care in the countries of the Americas, especially Chile. METHODS: The diagnoses and the patterns of use of mental health services were obtained from the Chilean Study of Psychiatric Prevalence (Estudio Chileno de Prevalencia Psiquiatrica). That representative research on the adult population of Chile was based on a stratified random sample of 2,987 people 15 years old and older, done over the period of 1992 to 1999. The psychiatric diagnoses were obtained using the Composite International Diagnostic Interview (CIDI). The interviewees were also asked about their use of general health care services and of mental health services in the preceding six months, as well as any barriers to accessing the services. RESULTS: More than 44% of the interviewees had had contact with some type of health service during the six months prior to the study, but only 5.6% received specialized care. Those who presented with a diagnosis of obsessive compulsive disorder or of panic disorder consulted more frequently, but not in specialized centers. Consumption of substances such as alcohol and drugs as well as antisocial personality disorder were associated with a low level of consultation. When asked about sources of assistance for mental health problems, the majority of the interviewees mentioned only the formal health care system. More than 75% of the interviewees said that they were satisfied or very satisfied with the care that they had received. CONCLUSIONS: Our results confirm the existence of a wide gap between the need for care and the treatment that is actually received. The informal and folkloric alternative resources (priests, family members, healers, herbalists, etc.) were used less frequently than is generally believed to be true. The indirect barriers of access to services- linked to a lack of knowledge and to stigma--were more frequent than were the direct barriers. A sizable number of people who did not have a positive diagnosis on the CIDI utilized mental health services. PMID- 16354424 TI - A window of opportunity: declining rates of hepatitis B virus infection among injection drug users in Rio de Janeiro, and prospects for targeted hepatitis B vaccination. AB - OBJECTIVES: To measure hepatitis B virus (HBV) infection rates among injection drug users in Rio de Janeiro, Brazil, and to report their knowledge of and attitudes toward hepatitis and HBV vaccination. METHODS: 609 injection drug users recruited in Rio de Janeiro between 1999 and 2001 answered a questionnaire and were tested for hepatitis B and other blood-borne infections. Questions covered sociodemographic information, alcohol and illicit drug consumption, drug injection and sexual practices, medical history, and knowledge about HIV, AIDS and viral hepatitis. RESULTS: The prevalence of HBV infection was 27.1%, with 3.4% of the sample positive for HbsAg (active infection) and 0.8% positive for anti-HBs (indicating previous HBV vaccination). Most interviewees (81.3%) were aware of at least one form of viral hepatitis and received information from many different sources. In agreement with laboratory findings, 96.7% of the interviewees stated they had never been vaccinated against hepatitis B, but almost all unvaccinated interviewees (97.8%) said they would volunteer to be vaccinated if HBV vaccination were available. CONCLUSIONS: Few of the injection drug users surveyed had ever been vaccinated against HBV. Although most were aware of the risks posed by viral hepatitis, this awareness seldom translated into consistent behavioral change. The participants' willingness to be vaccinated against HBV suggests that the implementation of vaccination for this population may help decrease rates of hepatitis B infection. PMID- 16354425 TI - [Evaluation of the Program for Depression Detection, Diagnosis, and Comprehensive Treatment in primary care in Chile]. AB - OBJECTIVE: To conduct a preliminary evaluation of the Program for Depression Detection, Diagnosis, and Comprehensive Treatment, which was developed by the Ministry of Health of Chile. MATERIAL AND METHODS: A group of women who had entered the Depression Program in eight primary care centers in three regions of Chile were monitored for three months. The initial sample was made up of 229 women, of whom 169 were able to be interviewed at the end of the three months. Sociodemographic variables, psychosocial risk factors, anxiety and somatic symptoms, and the degree of satisfaction with the care received were studied. Depressive symptoms were evaluated with the Beck Depression Inventory (BDI). The treatments prescribed for the women were also analyzed. RESULTS: The clinicians' decision to enroll the patients in the Depression Program was appropriate, with 95.2% of the women having depressive symptoms, according to the BDI. The accuracy in the clinical assessment of the severity of the symptoms was around 50%, with a tendency to underestimate the severity of the depression. The dropout rate for the women was 19.5% at the end of three months of monitoring. Those who dropped out tended to present less severe clinical symptoms (less intense anxiety and somatic symptoms), a smaller number of psychosocial risk factors, and a lower level of satisfaction with the care received. The patients showed good adherence (73.3%) with the pharmacological treatment, but less with the individual psychotherapy (47.4%) and the group psychotherapy (37.8%). A significant decline was seen in the intensity of the depressive symptoms by the end of the three months, with the decline being greater among the women with more serious symptoms. There was also a significant decline in anxiety and somatic symptoms. CONCLUSIONS: The preliminary evaluation indicates that the Depression Program is meeting the proposed objectives, although there appears to be a need to develop specific strategies to improve the accuracy of the clinical diagnosis as well as compliance with the treatments that are offered. PMID- 16354426 TI - Psychological and psychopathological reactions in Honduras following Hurricane Mitch: implications for service planning. AB - BACKGROUND: Posttraumatic stress disorder (PTSD) and other psychopathological outcomes have not been sufficiently studied in community-based samples in Latin America. This study explored various psychopathological reactions and their respective risk factors two months after Hurricane Mitch struck Honduras in October 1998. METHODS: In the Honduran capital of Tegucigalpa, 800 respondents age 15 and older were selected from residential areas of high, middle, or low socioeconomic status that had suffered either high or low impact from the devastating effects of the hurricane. The Composite International Diagnostic Interview was used to diagnose PTSD. Depression, alcohol misuse, and grief reaction were examined using screening instruments, and the Self-Reporting Questionnaire was used to measure demoralization. The Impact of Event Scale was administered to ascertain the severity of the posttraumatic reaction. RESULTS: PTSD was present in 10.6% of the sample. Respondents from the high-impact residential areas were more distressed, had higher scores on the grief inventory, and showed greater severity in PTSD symptoms. The respondents from the high impact residential areas also had higher prevalence rates of major depression, alcoholism, and prior emotional problems. The best explanatory model for the risk of developing PTSD included the degree of exposure based on reported traumatic events, and associated increased demoralization. Among the persons with PTSD, its severity was predicted by being female and by the degree of exposure to hurricane related traumatic events. CONCLUSIONS: Out of a total population of 3.3 million adults (age 15 and older) in Honduras, it is estimated that over 492,000 of them may have developed PTSD due to Hurricane Mitch. Adequate health disaster preparedness and response requires full acknowledgement of the multiple psychological effects that victims experience. PMID- 16354427 TI - [Managing epilepsy in the primary care network in Brazil: are health professionals prepared?]. AB - OBJECTIVE: To assess attitudes and beliefs concerning epilepsy held by physicians and allied health professionals who work in the primary care network in Brazil. METHODS: The data were collected during a national family and community medicine conference held in Rio de Janeiro in 2004, which was attended by 1,200 health professionals from throughout the country. Two questionnaires were used, one for physicians and another for nonphysician health workers. RESULTS: A total of 598 conference participants (345 physicians and 253 nonphysician health professionals) completed a questionnaire. Both the physicians and the nonphysician health professionals had acceptable personal attitudes towards persons with epilepsy. According to data from 286 physicians who provided information, the estimated mean percentage of individuals with epilepsy in the populations cared for by the physicians' health care systems in the preceding year was 0.78% (range, 0 to 8%; median = 0.37%). The estimated mean percentage of seizure-free patients was 60%. The estimated mean percentage of persons on monotherapy was 55%. The estimated mean percentage of referrals to a neurologist was 59%. The estimated mean percentage of patients who were working or studying was 56%. Of the 345 physicians, 252 of them (73%) reported not feeling confident about managing individuals with epilepsy, whereas 84 (24%) felt confident, and 9 (3%) were not sure. Of the 252 physicians who did not feel confident, 226 (90%) said that they would participate in a training program to improve the quality of the care provided to individuals with epilepsy. CONCLUSIONS: The professionals participating in the study indicated that the primary care system has the essential elements needed to provide comprehensive epilepsy care, including professional personnel who want to improve their skills. However, establishing an efficient, effective program of epilepsy management in the primary care network will require an effective referral network, regular distribution of antiepileptic drugs, the close monitoring of epilepsy management by using the federal primary health information system, and the ongoing education of health professionals, with emphasis on psychological support and the inclusion in society of individuals with epilepsy. PMID- 16354428 TI - Human immunodeficiency virus, AIDS, and drug consumption in South America and the Caribbean: epidemiological evidence and initiatives to curb the epidemic. AB - OBJECTIVE: The paper reviews data on drug use in relation to the spread of human immunodeficiency virus and AIDS in South America and the Caribbean. METHODS: Information was gathered by thoroughly reviewing major bibliographic databanks, web sites of international institutions and regional networks working with substance misuse or human immunodeficiency virus and AIDS, and abstracts from conferences and meetings. RESULTS: Although some gaps remain, a growing body of evidence documents the significant role of injected cocaine in the Brazilian and Southern Cone epidemics. The Caribbean and the Andean areas have thus far been spared in large part from the spread of injection drug use and its consequences, but the situation has been changing in Southern Cone countries towards a higher prevalence of harmful injection habits. Additional challenges have been posed by the increasing availability of heroin in the Andean Area and the abuse of crack cocaine and its impact on the sexual transmission of human immunodeficiency virus in many cities. Harm reduction strategies have been established in most areas of Brazil and are gaining momentum in Argentina. Other countries in the Region still face serious limitations due to restrictive legislation and lack of broader support. CONCLUSION: Greater participation of Latin American and Caribbean countries in research protocols and continued debate on both successful and failed experiences should be encouraged in order to minimize existing barriers to the full adoption of effective measures to curb the human immunodeficiency virus and AIDS epidemic in this Region. PMID- 16354429 TI - [Technical cooperation strategies of the Pan American Health Organization in the new phase of mental health services reform in Latin America and the Caribbean]. AB - The beginning of the new millennium coincided with the start of a new phase in the reform of mental health services in Latin America and the Caribbean. This new phase has imposed new priorities and prompted new technical cooperation strategies at the international level. This piece points out the main characteristics of the first phases in the reform of mental health services in Latin America and the Caribbean, discusses the factors that led to the phase that started in 2001, and describes the strategies and the technical cooperation activities of the Pan American Health Organization to deal with the challenges that have arisen in the current stage of reform. The piece also considers the prospects for international cooperation in this field, as well as the advantages of establishing a program for the reform of mental health services in the Americas that would contribute to the combined efforts of governments and international organizations in an action plan with defined objectives. The piece recommends taking advantage of the celebration of the 15th anniversary of the Declaration of Caracas in order to launch an action plan that gives new impetus to mental health services reform in the Americas. PMID- 16354430 TI - Evidence-based alcohol policy in the Americas: strengths, weaknesses, and future challenges. AB - The objectives of this article are to describe the evidence base for alcohol policy in the Americas, to evaluate the extent to which national policies are likely to have an impact on public health, and to identify areas where alcohol policies could be improved. The paper begins with a brief review of epidemiological surveys of the prevalence of alcohol problems in the Americas. This is followed by an analysis of 32 prevention strategies and interventions in terms of the evidence for their effectiveness, amount of research support, cost to implement, and other feasibility issues. Overall, the strategies and interventions with the greatest amount of empirical support are low blood alcohol concentration levels for driving while intoxicated, controls on alcohol availability, age limits on alcohol purchases, and relatively high alcohol prices. The implications of the evidence are next discussed in relation to alcohol policy initiatives in the Americas, based on an analysis of the extent to which strategies and interventions currently used in 25 countries of the Americas are likely to have a public health impact on alcohol-related problems. The countries that have adopted the policies with the highest expected impact overall are Colombia, Costa Rica, Venezuela, and El Salvador. Nevertheless, the analysis indicates that almost all the countries of the Americas could improve the likelihood of preventing alcohol-related problems. Policy efforts in the developing countries of Latin America should focus on improving countermeasures against driving while intoxicated, measures that alter the drinking context, and limits on physical availability. For the developed, high-income countries of North America the goal should be to prevent deterioration of current drinking patterns and to reduce the overall volume of drinking. Given the low to moderate cost of many of the policies reviewed in this article, it now seems possible for communities and nations to substantially reduce the alcohol-related burden of illness in the Americas. PMID- 16354431 TI - An integrated program to train local health care providers to meet post-disaster mental health needs. AB - This paper describes a post-disaster mental health training program developed by the International Section of the Department of Psychiatry at Dalhousie University (Halifax, Canada) and delivered in Grenada after Hurricane Ivan struck the country in September 2004. This train-the-trainer program used an integrated community health model to help local health care providers develop the necessary skills for the identification and evidenced-based treatment of mental disorders occurring after a natural disaster. The approach also provided for ongoing, sustainable mental health care delivered in the community setting, as advocated by the World Health Organization and the Pan American Health Organization. This approach is in contrast to the largely ineffective and costly vertical whole population psychosocial counseling activities that have often been used in the Caribbean following natural disasters. PMID- 16354432 TI - [The National Mental Health Plan in Chile: 10 years of experience]. AB - Dealing with mental health problems is undoubtedly an increasingly important public health responsibility around the world. In Chile, because of the changes in the epidemiological profile of the population, the lifetime prevalence of mental and behavioral disorders has reached 36%. In response, the Ministry of Health of Chile, through its Mental Health Unit, prepared the National Plan for Mental Health and Psychiatry. The Plan establishes objectives, strategies, and steps to improve the well-being and mental health of Chileans. This piece describes the model of care for mental health and psychiatry used in Chile's public health care system, analyzes the main difficulties encountered and the achievements made in the 10 years that the Plan has been in place, and makes recommendations for improving the Plan. Over the 10-year period, the new model for mental health and psychiatry has managed to make a place for itself in the public health care system. Indicators show that the beneficiaries of the public health care system in Chile now have greater access to mental health services than before the new model of community care was established, have broader health care coverage, and receive better quality services. PMID- 16354433 TI - Child and adolescent mental health in Latin America and the Caribbean: problems, progress, and policy research. PMID- 16354434 TI - Mental health and due process in the Americas: protecting the human rights of persons involuntarily admitted to and detained in psychiatric institutions. PMID- 16354435 TI - Human rights and deinstitutionalization: a success story in the Americas. PMID- 16354436 TI - How to address patients' defences: a pilot study of the accuracy of defence interpretations and alliance. AB - This pilot study examined the accuracy of therapist defence interpretations (TAD) in high-alliance patients (N = 7) and low-alliance patients (N = 8). TAD accuracy was assessed in the two subgroups by comparing for each case the patient's most frequent defensive level with the most frequent defensive level addressed by the therapist when making defence interpretations. Results show that in high-alliance patient-therapist dyads, the therapists tend to address accurate or higher (more mature) defensive level than patients most frequent level. On the other hand, the therapists address lower (more immature) defensive level in low-alliance dyads. These results are discussed along with possible ways to better assess TAD accuracy. PMID- 16354437 TI - Psychological dynamics affecting traumatic memories: implications in psychotherapy. AB - The search to understand response to trauma has turned to the contribution of personality factors. The way people process the stressor event is critical in determining whether a trauma will be configured or not. Neuroscience shows that the brain does not store memories, but traces of information that are later used to create memories, which do not always express a completely factual picture of the past experience. Whenever an event is retrieved, it may undergo a cognitive and emotional change. Psychological dynamics--emotional interpretative tendency that affects the internal dialogue related to a meaningful event--may influence the development of positive or negative outcomes after stressor events. We postulate that therapists must see beyond the traumatic event itself and work with the internal dialogues that maintain the pathological relationship with the past episode. Thus, they may better treat traumatized patients by therapeutically rebuilding the memory. A brief clinical case is presented to show how exposure based and cognitive restructuring therapy may help trauma victims experience psychological growth from their negative experiences, by fostering healthy psychological dynamics. PMID- 16354438 TI - Ripley's Game: projective identification, emotional engagement, and the counselling psychologist. AB - Counselling psychology's dual emphasis on the use of the self as both vehicle of therapeutic change and legitimate focus of inquiry and research is one of the features by which the profession may be distinguished from related fields such as clinical psychology or psychotherapy. This paper discusses the relevance of the psychoanalytic concept of projective identification in understanding the extent, nature and subtlety of the ways in which the therapist's 'self' and emotions may be deployed within the therapeutic relationship. Illustrated with reference to the film 'Ripley's Game' and a clinical case vignette, Searles's (1978) advocacy of 'a richness of emotional participation' within clinical work is discussed in relation to the pluralist philosophy of counselling psychology; implications for the training and personal development of counselling psychologists are also briefly explored. PMID- 16354439 TI - A prospective study of the effectiveness of group and individual psychotherapy for women CSA survivors. AB - OBJECTIVES: There is little empirical evidence to guide clinical practice in treating adult patients presenting to adult mental health and primary care services with severe psychological difficulties consequent on childhood sexual abuse. The aim of this study was to determine the effectiveness of a model of short term, focal, integrative psychotherapy with this population of patients and to compare outcomes when the model is delivered on an individual or group basis. DESIGN: This study used a patient preference design with random allocation to one of two treatment modalities (individual or group treatment). There was a waiting list control. METHOD: A group of 48 women patients were assessed on 4 psychological measures when entering a waiting list condition, immediately before treatment and after completion of 12 sessions of psychotherapy, either in a group or individually. Follow-up data were collected at 4 months and 8 months. RESULTS: Both individual and group patients showed highly statistically and clinically significant improvements after treatment. These gains were maintained at follow up with the exception of one measure that indicated a significant decline from post-treatment levels for the group patients. CONCLUSIONS: This model of psychotherapy is highly effective, when delivered by chartered clinical psychologists to patients whose preferences for individual or group therapy have been met. PMID- 16354440 TI - Characteristics of participants in the Forum, psychotherapy clients, and control participants: a comparative study. AB - 'New age' activities, personality variables, symptomatology, and subjective well being (SWB) among the Forum (F) participants, psychotherapy (P) clients, individuals who are both Forum and psychotherapy (FP) clients, and control (C) participants, who were never involved in either F or P were compared. A group of 64 Israeli men and 76 women (mean age = 38.55 years) completed a demographic questionnaire; the Internal-external, Sensation-seeking, Happiness, Affects balance, Satisfaction with life, and Anxiety and depression scales of the SCL-90. The FP and F participants used occult counsellors significantly more than the C participants, although gender differences were also found. All in all, the FP and P clients were more depressive and anxious, and less happy, less satisfied with their life, and affectively balanced than the F and the C participants. The rationality, personal responsibility, and mental health of the F participants are discussed in light of the results with considerations of short-versus long-term interventions. PMID- 16354441 TI - Negotiating a pathological identity in the clinical dialogue: discourse analysis of a family therapy. AB - Within the framework of social constructionism, psychotherapy has been re conceptualized as a semiotic process, which consists of the creative generation of new meanings in the context of collaborative discourse. In recent years, research approaches that draw from social constructionism, such as discourse analysis, have been fruitfully employed in the study of psychotherapy processes, whilst being in line with the contemporary emphasis on language, narrative, and meaning making. This paper aims to further the exploration of the usefulness of discourse analysis in the study of psychotherapy processes, and in particular, in situations where the medical discourse is powerfully implicated in the construction of a person's identity. It is based on the analysis of a family therapy with a family whose child has a diagnosis of autism. The analysis focuses on two features of the family's talk, namely shifts in the flexibility of employment of a diverse range of discourses and subject positions, and shifts in the ways agency is constructed and discursively negotiated in the clinical conversations. It is suggested that these shifts can be used as indications of change in the family's network of meanings. The analysis suggests that an important aspect in clinical work with families with a member with a psychiatric diagnosis lies in decentring, or deconstructing, the dominant, pathology maintaining accounts, and allowing for a wider range of less problematic narratives and subject positions to emerge. PMID- 16354442 TI - Factors in outcome of cognitive-behavioural therapy for psychosis: users' and clinicians' views. AB - OBJECTIVES: Despite increasing evidence to suggest that cognitive-behavioural therapy (CBT) is helpful for a significant proportion of people with psychosis, only limited information is available regarding factors implicated in outcome. The present study investigated factors differentiating outcomes on the basis of accounts from participants in the therapeutic process. METHOD: Four therapists and eight of their clients were interviewed about their experiences of CBT. Clients were defined as having progressed or not progressed during therapy. Interview data were analysed using a qualitative 'grounded theory' methodology. RESULTS: A number of major categories differentiated the two client groups, including ability to let go of distressing beliefs, logical thought, holding therapy, and presence of a shared goal. Overall, clients who progressed were better able to move into the therapist's frame of reference. Therapists and clients also felt that non-specific benefits accrued from the therapy for both groups. CONCLUSION: The results were consistent with previous studies suggesting that ability to disengage from distressing beliefs is important in therapeutic progression. Reasons considered for the inability to progress include emotional investment in psychotic beliefs and cognitive processing. Further research is required to clarify the role of logical thought and therapeutic alliance in progress and in predicting outcome. PMID- 16354443 TI - Utilization of psychotherapy in patients with personality disorder: the impact of gender, character traits, affect regulation, and quality of object-relations. AB - OBJECTIVES: The aim of the study was to generate hypotheses for examining gender differences in variables with predictive value for the utilization of psychotherapy in patients with personality disorders (PDs). DESIGN: Personality traits, affect experience and regulation, the quality of object relations and interpersonal problems within the process of psychotherapy planning were assessed in 140 psychiatric outpatients. METHODS: Besides the structured clinical interviews for DSM-IV I+II, variables were assessed with the Shedler-Westen assessment procedure (SWAP-200), the affect regulation and experience Q sort (AREQ), the quality of object-relations scale (QORS), and the Inventory of Interpersonal Problems (IIP). Correlation and group difference statistics, regression and canonical correlation analysis were performed. RESULTS: Predictors concerning the utilization or non-utilization of psychotherapy were a schizoid PD rating a self-report of subassertive behaviour related to interpersonal problems in women, and a narcissistic PD rating in men. Canonical correlations between predictors and quality of object relations or interpersonal problems were found in women, while in men there was merely a tendency for predictor and affect regulation to be related. CONCLUSION: The results suggest that for men it is more important to interpret the dominating affect, while for women, understanding the pathological object relation pattern is useful for successful therapy planning. PMID- 16354444 TI - Early maladaptive schemas and adult attachment in a UK clinical population. AB - OBJECTIVES: Both insecure attachment style and early maladaptive schemas have been associated with mental health difficulties. This study aimed to investigate how schemas relate to attachment style classification in mental health service users. A further aim was to investigate the nature of the relationship between psychological distress, common psychological difficulties, and attachment style. METHOD: Participants were 72 people in contact with mental health services who completed the short form of Young's schema questionnaire, a measure identifying their attachment style (experience of close relationships), and the clinical outcomes in routine evaluation (CORE; Evans et al., 1998) outcome measure. RESULTS: Overall, 81% of the participants had an insecure attachment style. The fearful group were the most distressed across several domains of the CORE, followed by the preoccupied group. Schemas differed significantly according to attachment style grouping, with the fearful group possessing the greatest degree of maladaptive schemas, followed by the preoccupied group. Discriminant function analysis identified differing patterns of schemas associated with attachment style. CONCLUSIONS: The results supported the relevance of both attachment style and maladaptive schemas to individuals with mental health difficulties. Importantly, several aspects of the individuals' symptoms and difficulties and their schemas were meaningfully related to their adult attachment style. Limitations included the small number of dismissing individuals identified, the problems of self-report in this area, and the absence of information about the quality of participants' current relationships. PMID- 16354454 TI - Diet and healthy ageing. AB - In the future there will be more people aged 65 years and over ('older adults'). Although the exact mechanisms underlying normal ageing are not fully understood, ageing is generally associated with an increase in chronic diseases, such as cardiovascular disease, diabetes, cancer and osteoporosis. It is becoming clear that it is possible to prevent, slow or reverse the onset of many these by modifying lifestyle factors such as diet. Studies of older adults in a range of countries have highlighted a number of areas in which dietary quality could be improved. It is important to identify dietary patterns in addition to specific dietary components that offer protection against chronic disease. The challenge in the area of diet and healthy ageing is twofold: first, there is a need to improve the diet of older adults; and second, as most chronic diseases begin earlier in life, there is a need to encourage other age groups to adapt their diet so they can enter old age in better health. PMID- 16354455 TI - Reproduction and ovarian ageing. AB - Female fertility rates are inherently linked to a woman's age, which is in turn related to ovarian function. Reproductive potential declines gradually until 37 38 years of age, from when the rate of decline hastens. Approximately 1% of women suffer from premature ovarian ageing, and many may not have completed their families. This paper reviews the physiology and fertility consequences of ovarian ageing, premature ovarian failure, measures of ovarian reserve and methods of fertility preservation. PMID- 16354456 TI - The investigation and management of interstitial cystitis. AB - Interstitial cystitis (IC) is a chronic inflammatory disorder of the bladder that is notoriously difficult to manage and can result in considerable morbidity. It very likely overlaps with painful bladder syndrome, but they are different conditions. The aetiology remains obscure, and the definition and diagnostic criteria are debated. The diagnosis of IC is one of exclusion, frequently based on symptoms and cystoscopy findings. Typical symptoms include frequency, urgency, dysuria and lower abdominal, bladder, vaginal, urethral or perineal pain, in the absence of bacterial cystitis. Voiding often relieves the suprapubic discomfort, and drinking alcohol- and caffeine-containing drinks frequently exacerbates it. Many treatments have been tried, with little sustained success. Proposed systemic treatments include anti-histamines, heparin, amitriptyline and pentosan polysulfate (a synthetic analogue of glycosaminoglycan which augments the mucous protective layer of the bladder). In many patients symptoms are improved following cysto-distension but the benefits are short-lived. Instillations of dimethyl sulfoxide, hyaluronic acid or chondroitin also show promise. Where treatments have failed and symptom severity is such that the patient's quality of life is poor, a urological opinion should be sought and reconstructive surgery considered. Available options include partial cystectomy, augmentation cystoplasty, and urinary diversion with or without cystectomy. PMID- 16354457 TI - Is there a role for hormone replacement therapy after breast cancer? AB - Breast cancer is the most common female malignancy in the UK, with an overall lifetime risk of 1 in 9. Despite the high incidence, breast cancer mortality is decreasing. Approximately 40,000 women were diagnosed with breast cancer in England and Wales in 2000 but the majority will have normal or near-normal life expectancy. One of the main contributory factors to this marked improvement in survival over the last 20 years in women of all ages has been the more widespread use of systemic therapy in early-stage disease. For women with hormone-sensitive cancer, this involves adjuvant endocrine therapy that reduces estrogen synthesis (i.e. ovarian suppression in premenopausal women or aromatase inhibitors in postmenopausal women) or estrogen activity (the anti-estrogen tamoxifen, irrespective of menopausal status). Many women experience health and quality-of life problems related to estrogen deficiency as a result, the commonest being vasomotor symptoms and vaginal dryness. This article summarizes and interprets key recent papers on the use of hormone replacement therapy (HRT) and selective serotonin reuptake inhibitors in breast cancer survivors. HRT may be safe in women with receptor-negative disease or receptor-positive cancers in the presence of tamoxifen. However, there is a dearth of useful alternatives. PMID- 16354458 TI - Decade-long use of continuous combined hormone replacement therapy is associated with better health-related quality of life in postmenopausal women, as measured by the generic 15D instrument. AB - OBJECTIVE: To examine quality of life after a decade of continuous combined hormone replacement therapy. STUDY DESIGN: The 15D, a generic health-related quality-of-life (HRQoL) instrument, was used to evaluate trends in HRQoL in women who used continuous combined hormone replacement therapy (ccHRT; Indivina, Orion Pharma, Finland) for up to nine years. These women had a mean age of 56 years of age at the start of therapy. Control data on HRQoL were obtained from age-matched women participating in Finnish population health surveys. RESULTS: Relative to controls, ccHRT was associated with significantly better HRQoL after six and nine years of treatment. Dose minimization at 8.5 years was not associated with a decline in HRQoL in the ensuing six months. One year after discontinuation of ccHRT there was evidence of a decline in HRQoL in women who discontinued ccHRT as planned, whereas HRQoL was maintained in women who had continued or resumed ccHRT during the one-year post-study follow-up. The benefits of ccHRT were apparent in multiple dimensions of the 15D, being largest and most robustly reproduced in the dimension 'discomfort and symptoms'. It is conjectured that the effectiveness of ccHRT in relieving symptoms of menopause may have contributed to the improved HRQoL scores registered in other dimensions, in addition to any direct effects of ccHRT on specific aspects of those other dimensions. CONCLUSIONS: These data indicate that up to 10 years of low-dose ccHRT has sustained value in the enhancement of HRQoL when used by women for whom relief of symptoms of menopause and control of bleeding are primary objectives of treatment. PMID- 16354459 TI - Managing the menopause: British Menopause Society Council consensus statement on hormone replacement therapy. AB - The British Menopause Society Council aims to help health professionals inform and advise women about the menopause. This guidance regarding estrogen-based hormone replacement therapy (HRT), including tibolone, which is classified in the British National Formulary as HRT, responds to the results and analysis of the randomized Women's Health Initiative studies and the observational Million Women Study. Treatment choice should be based on up-to-date information and targeted to individual women's needs. HRT still offers the potential for benefit to outweigh harm, providing the appropriate regimen has been instigated in terms of dose, route and combination. PMID- 16354460 TI - Premature menopause: management challenges after the Women's Health Initiative. PMID- 16354461 TI - Pharmacotherapy of female urinary incontinence. AB - Urinary incontinence is a major clinical problem and a significant cause of disability and dependency in older adults. Overall, the prevalence of urinary incontinence approaches 55% among women aged over 55 years. The past few years have seen significant advances in the pharmacotherapy of overactive bladder and stress incontinence. The review examines the evidence regarding their benefits and side-effects. PMID- 16354462 TI - Hormone replacement therapy and colorectal cancer: an update. AB - Observational studies and randomized clinical trials that have looked at the effect of hormone replacement therapy (HRT) on the risk of colorectal cancer are reviewed. Nine cohort studies in this area have included a total of over 2700 cases of colorectal cancer. Most of these studies found a relative risk (RR) of around or below unity. Of 15 case-control studies, with a total of over 7300 cases, six reported a 20-40% risk reduction among women who had ever used HRT. With reference to randomized clinical trials, in the Women's Health Initiative (WHI) study, after seven years of follow-up, 45 cases of colorectal cancer were observed in the combined HRT group versus 67 in the placebo group, corresponding to a RR of 0.63. A combined reanalysis of data from both the WHI and the Heart and Estrogen Progestin Replacement Study (HERS) included 56 cases in the combined HRT group and 83 cases in the placebo group (pooled RR = 0.64). However, in the WHI study cancers diagnosed in the HRT group were more advanced and there were more positive lymph nodes. Furthermore, among women in the WHI who had had a hysterectomy, there was no difference at the eight-year follow-up in the incidence of colorectal cancer between those in the estrogen-only arm of the trial (n = 61) and those in the control group (n = 68). PMID- 16354463 TI - Reflections on the impact of the Bamako Initiative and the role of user fees. PMID- 16354464 TI - Does maternity care improve pregnancy outcomes in women with previous complications? A study from Zimbabwe. AB - To determine the utilization of maternal health care services and pregnancy outcomes for women with a history of complications in previous pregnancy, we analysed the pregnancy records of multiparous women (parity > or =1) who booked and completed follow-up in Gutu district, Zimbabwe between January 1995 and June 1998. Women with previous uncomplicated pregnancies (n = 6140) were classified as low risk, whereas those with complications of previous pregnancy (n = 1077) were classified high risk. At enrolment, there was no difference in maternal age and parity between low- and high-risk women. A higher proportion of high-risk women had more than five antenatal visits (32% versus 21%; P<0.001) and gave birth in hospital (47% versus 18%; P<0.001). The risk of antenatal (relative risk [RR] 1.57; 95% confidence interval [CI] 1.32-1.88), labour/delivery (RR 1.98; 95% CI 1.75-2.25) and neonatal (RR 1.83; 95% CI 1.44-2.34) complications was elevated in high-risk women. There was increased risk for perinatal death in high-risk women, but this did not reach statistical significance (RR 1.56; 95% CI 0.98-2.49). The recurrence ratio for most complications was low and the sensitivity of historical risk markers in predicting women likely to develop further complicated pregnancies was only 23%. Most women with previous pregnancy complications can safely give birth in the rural health centre. We concluded that high-risk women had an elevated risk of complications in the index pregnancy and that better utilization of maternal health care, especially for delivery, reduced adverse perinatal outcomes. PMID- 16354465 TI - Cervical spine injuries in 64 attempted suicidal hangings in India. AB - This report describes the outcome of 64 attempted suicidal hangings, 81% of whom made a full recovery. PMID- 16354466 TI - Choice of drainage procedure in paediatric pyopericardium: a 30-year experience. AB - Although there is general agreement on the necessity of draining pyopericardium, debate continues as to the safe and effective method of drainage. Studies describing head-to-head comparison of various drainage procedures are very few and are disadvantaged by small numbers of cases. In this observational study, we review our 30-years experience with different techniques of pericardial drainage. Between 1972 and 2003, the authors have personally treated 39 children who suffered from pyopericardium. Among the 22 children who underwent early partial pericardiectomy, 20 were alive. In contrast to this, 12 out of 15 children treated with repeated pericardiocentesis or sub-xiphoid tube drainage were dead. The median hospital stay for pericardiectomy group was 18 days (range 11-32) and that for the non-thoracotomy group was 34 days (range 18-55 days). With regard to immediate survival and early convalescence in the pyopericardium, partial pericardiectomy is superior to pericardiocentesis and sub-xiphoid tube drainage. Pericardiocentesis can be used for diagnostic or temporizing purposes, but not as the definitive drainage procedure. Partial pericardiectomy can be done even in small hospitals where heart-lung machines are not available. PMID- 16354467 TI - Determining the sensitivity of sitting height in predicting cephalo pelvic disproportion in Eritrea. AB - To determine the sensitivity of sitting height in predicting cephalo pelvic disproportion (CPD), a prospective case-control study was conducted. A total of 724 primiparas who delivered in Mekane Hiwot Maternity Hospital, Asmara, Eritrea were studied. Primiparas who delivered by caesarian section because of CPD were taken as cases (221), and primiparas who delivered vaginally were taken as controls (503). Height was measured for all, while sitting and while standing. The sensitivity of sitting height in predicting CPD was 34.1% (95% confidence interval [CI] = 27.7-40.3%), while that of the standing height was 33.5% (95% CI = 27.1-39.84). CPD cannot be predicted accurately by height in general and by sitting height in particular. Therefore, in order to avoid obstetric complications, all pregnant women should have access to skilled attendance during delivery and access to facilities with emergency obstetric care when needed. PMID- 16354468 TI - Epidemiological study of patients with connective tissue diseases in Brazil. AB - To study the prevalence of connective tissue disorders and identify the epidemiological profile of the population from the northern and northeastern regions of Brazil. All patients admitted to the general medical ward at Getulio Vargas Hospital were screened through history and physical examination for the existence of known, suspected or possible connective tissue disorder. A total of 120 patients were enrolled in the study - 61 (50.8%) with systemic lupus erythematosus (SLE), 41 (34.2%) with rheumatoid arthritis (RA), 16 (13.3%) with systemic sclerosis (SS), one (0.8%) with mixed connective tissue disease (MCTD) and one (0.8%) with polymyositis (PM). Women accounted for (104) 86.7% of the participants and (83) 70.9% were non-Caucasians. The mean age was 36.8 years. Alopecia was the most frequent symptom (55/46.6%), followed by other cutaneous lesions (50/42.4%), dyspnea (47/39.8%) and chest pain (39/33.0%). The most frequent rheumatologic disorder encountered in our hospital-based study population was systemic lupus erythematosus, which is contrary to findings of the majority of studies performed in this country. PMID- 16354469 TI - Comparison of three different combination therapies in the treatment of human brucellosis. AB - The efficacy and tolerability of three different combination treatment regimens in human brucellosis were compared in 118 uncomplicated patients enrolled in a prospective study between May 1997 and December 2002. Brucellosis was diagnosed using standard clinical and microbiological findings. Patients with central nervous system involvement, spondylitis, endocarditis or children under 16 years of age were excluded from the study. Patients were randomly assigned to receive 400 mg of ofloxacin plus 600 mg of rifampicin (OR, n = 41), 200 mg of doxycycline plus 600 mg of rifampicin (DR, n = 45) or 1g intramuscularly streptomycin (administered for three weeks) plus 200 mg doxycycline (DS, n = 32) daily for 6 weeks. All patients were followed up at least 6 months after cessation of therapy. There was no statistical difference between the groups on relapse rates and clinical response to the treatment (P>0.05). Five patients in OR (12.8%), six patients in DR (14.3%) and three patients in DS groups (9.7%) suffered relapse. The side-effects were seen in eight (19.5%), 21 (46.7%) and eight (25.0%) patients of OR, DR and DS groups, respectively. The use of combination therapy of ofloxacin plus rifampicin for 6 weeks was found to be as effective as DR and DS. The side-effects of therapy in OR and DS groups was less severe than in the DR group. PMID- 16354470 TI - Comparison of albendazole regimen for prophylaxis of strongyloides hyperinfection in nephrotic syndrome patients on long-term steroids in Cambodia. AB - Nephrotic syndrome patients on long-term steroids face the risk of having heavy uncomplicated strongyloidiasis or death from its extreme form, the strongyloides hyperinfection. The risk can be minimized if we eradicate the parasite first. We compare a once daily and twice daily albendazole regimen in preventing this potentially fatal complication in 122 patients with nephrotic syndrome. PMID- 16354471 TI - Sexual risk behaviour among young people in Osogbo, Osun state Nigeria. AB - By analysing data randomly collected via administration of questionnaire by interview of individuals on sexual risk behaviour to 628 young people living in Osogbo, Nigeria, the results of the questionnaire revealed that the mean age of first sexual encounter was 17.4 years for men and 16.5 years for women. Men tended to have multiple sexual partners more than the women (74.4% and 25.1%, respectively), while there was a highly significant relationship between sexually transmitted disease and multiple sexual partners among singles with P<0.001 and P= 0.000. We concluded that parents and government alike should teach sex education at home, and in schools involving intensive counselling. PMID- 16354472 TI - Assessment of physical conditions and current practice in laboratories carrying out sputum smear microscopy in Northwest Ethiopia. AB - A cross-sectional study was conducted in Northwestern Ethiopia involving 18 health-care institutes in August 2003 to assess the physical conditions and current laboratory practice, pertaining to handling sputum specimens in the health-care laboratories carrying out sputum smear microscopy. A structured and pre-tested questionnaire was used to collect information. The laboratories had an area of less than 25 m(2). None of them had separate rooms or safety cabinets for sputum smear preparation. Only three laboratories used facemasks. QJ;Decontamination of sputum specimens prior to disposal was reported in only two. Incineration as a means of sputum specimen disposal was used in only six laboratories. The physical conditions of the laboratories were found below the standard set by the International Union Against Tuberculoses and Lung Diseases. A large number of sputum specimens continue to be handled, despite the poor conditions and procedures in all the healthcare laboratories. Laboratory safety standards need to be improved, to minimize the risk of infection. In addition, there is an urgent need for education, training and supervision of the staff involved in sputum microscopy. PMID- 16354473 TI - Assessing traditional healers: an observational clinical study of classical Arabic medicine in Mauritania, with comparison of prognosis and outcome. AB - In a classical Arabic medicine clinic near Nouakchott, Mauritania, two general practitioners observed 170 consecutive consultations. With a follow-up rate of 66%, the studied practices appeared fairly safe and, in many chronic conditions, traditional healers performed similarly to what can be expected with modern medicine. Traditional healers and professionals from the nearby primary health care centre started a weekly joint consultation. PMID- 16354474 TI - Incidence of fungi pathogens associated with root canal treatments, in Benin City, Nigeria. AB - In order that appropriate clinical interpretation of microflora and drug management of root canal infections would be achieved, the fungi contents of root canal following biomechanical instrumentation were determined. Mucor, yeast, candida, microsporium and fusarium species were recovered at the different stages of instrumentation. The presence of fungi could complicate root canal treatments. The possible implications of these findings in treatment failures are highlighted. PMID- 16354475 TI - Deaths associated with anaesthesia in Togo, West Africa. AB - This study attempts to determine the anaesthetic death rate, the causes of deaths and the avoidable mortality rate (AMR) in consecutive cases. The number of anaesthetics given was 1464: 30 cases died within 24 h. The incidence of 24-h perioperative deaths per 100 anaesthetics was 2.57. In all, 50% of deaths were observed in obstetric surgery; 47% of deaths were associated with cardiovascular management, 30% with respiratory management; 93% of deaths were identified as avoidable. The AMR was 1.5% (anaesthetic AMR: 0.75%, administrative AMR: 0.68%, surgical AMR: 0.07%). Insufficient or no blood available is the only factor for administrative AMR. PMID- 16354476 TI - A survey of the use of medicinal plants and other traditional medicine in Kasese District, Uganda. AB - The use and value of traditional plants and medicines is only slowly being investigated by Western medical organizations. A survey of 492 members of support groups and health-care clinics in Kasese district, Uganda was undertaken in a group setting: 23 groups with a mean size of 21, age range 4-53, which represented 0.1% of the population, covering nine of the 20 sub-counties, both in the native languages and in English, using photographs and specimens of 12 plants. Most admitted to using plants at some time-81% for self, 77% for their children; 45% admitted to using traditional healers as a source for information about health. Most plants were home grown or available locally. Medicinal plants were used for respiratory infections, fever, malaria and diarrhoea/vomiting. HIV/AIDS was rarely treated with medicinal plants. PMID- 16354477 TI - Issues of measuring and improving the treatment of malaria in sub-Saharan Africa. AB - This paper, using focus group discussions and questionnaires in Enugu State, Southeast Nigeria, examines the implications of consumer malaria perceptions and behaviour for measuring the disease burden and improving its treatment. The results show that, because peoples' understanding of the disease was related to its symptoms, this could lead to overestimation of the economic burden of malaria, based only on surveys without diagnostic confirmation. Survey-based estimations of the burden of malaria should control for the different local terminologies of malaria, and health personnel should be aware of these in order to improve the appropriate use of antimalarial drugs in presumptive treatment of malaria. PMID- 16354478 TI - Drug use among adolescents in Ilorin, Nigeria. AB - The types and frequency of drug use among 1200 students aged 10-19 years was investigated and a prevalence rate of 40.1% found; currently used drugs included mild stimulants such as kolanut and coffee 294 (26.2%), alcohol 164 (14.5%), sniffing agents 80 (7.2%), amphetamine and ephedrine 66 (6.7%), cigarette 54 (4.8%), heroin 45 (4%) cocaine 40 (3.6%) and cannabis 38 (3.4%). Multiple drug use was found among the students, with the abuse of cannabis, cocaine and heroin being significant among those who smoked cigarette (P<0.001). The relative risk (RR) for cannabis use when cigarette was smoked was 37.4 (24.1-57.8); RR for cigarette smoking when alcohol was used, 6.8, while RR for cocaine abuse when cigarette was used, 21.8 (13.9-34.5) and 52.8 (29.2-95.5) when cannabis was used. It is therefore concluded that the use of the licit and "socially" acceptable drugs may pave the way for the abuse of illicit ones. PMID- 16354479 TI - Childhood brucellosis in Babol, Iran. AB - This prospective study was conducted on 111 children with brucellosis, who attended the Department of Infectious Diseases, Babol, Iran, from September 1999 to March 2003. Fever (73.9%), sweating (67.6%), artheralgia (49.5%) and splenomegaly (18%) were common findings. The disease was acute and sub-acute in 97.3% and chronic in 2.7% of cases. Peripheral arthritis was seen in 35 (31.5%) of cases, which was monoarticular in 29 patients, most involving the knees and hips. Sacroilitis was seen in six (5.4%) cases. PMID- 16354480 TI - Aetioepidemiological profile of spinal injury patients in Eastern Nepal. AB - This is a retrospective case series of 233 spinal injury patients admitted to the orthopaedic ward of BPKIHS from May 1997 to April 2001. The inpatient records were analysed. In all, 40.3% of spinal injuries resulted from falls from trees while cutting leaves for fodder, and 27.9% resulted from falls from first/second floors. More than 75% of total spinal injuries are largely preventable. Overall, 46.8% of our spinal injury patients had complete cord transection at the level of injury. All adolescents and adults, irrespective of age or sex, should be the target groups for community education and intervention programmes for prevention of spinal injury. PMID- 16354481 TI - Unusual sites of hydatid cysts in India. AB - Hydatid disease usually affects liver and lungs, but may affect any organ, posing a diagnostic and therapeutic dilemma. We analysed 110 patients with hydatid cyst over 21 years in our general surgical unit, which included 24 cases in unusual sites. The spleen was the most common, followed by skin and soft tissues. PMID- 16354482 TI - Puffer fish poisoning (tetrodotoxin) in Bangladesh: clinical profile and role of anticholinesterase drugs. PMID- 16354483 TI - Obstructed labour due to sacral TB. PMID- 16354484 TI - Generalized tetanus complicating lower limb fractures managed by traditional bone healers. PMID- 16354485 TI - Intramuscular ketamine anaesthesia for paediatric eye surgery in Madagascar. PMID- 16354486 TI - Splenic abscess in typhoid fever. PMID- 16354487 TI - Guinea worm infection of urinary bladder manifesting as obstructive uropathy in rural Maharashtra. AB - Guinea worm or Dracunculus medinensis is a well-documented helminthic infestation in many areas of Asia. In this report, we describe a rare case of guinea worm infestation in a 25-year-old woman who had developed symptoms of obstructive uropathy, in whom fragments of guinea worm were removed after urethral catheterization. To the best of our knowledge, adult guinea worm occurring in the urinary bladder has not been previously described. PMID- 16354488 TI - Cholecystitis and cholelithiasis in adults in Zaria. PMID- 16354489 TI - Taenia-induced ileal perforation. AB - An unusual case of taenia-induced ileal perforation is reported and the management is discussed. PMID- 16354490 TI - Complications from traditional tooth extraction in South-western Uganda. AB - We report on the practices of traditional tooth extraction and highlight the complications. PMID- 16354491 TI - Isolated splenic TB report of three cases. PMID- 16354492 TI - Peritonitis in patients over the age of 50 years: 98 cases managed surgically. AB - Generalized peritonitis in patients over the age of 50 years is a common surgical emergency. This is a retrospective analysis of 98 cases managed surgically. Duodenal ulcer perforations, necrotizing enteritis, acute cholecystitis with perforation and small bowel perforations were the common causes. Most of them presented late, and many had associated conditions. Re-look laparotomies had a definite role to play. While there is significant decrease in the number of typhoid and tubercular peritonitis, there appears to be an increase in the incidence of necrotizing enteritis and acute cholecystitis. PMID- 16354493 TI - Offering highly active antiretroviral treatment to children in Thailand: the difficulties. PMID- 16354495 TI - Causal hypothesis for some congenital anomalies. AB - Congenital anomalies are a major cause of fetal and neonatal death and of childhood morbidity. Chromosomal and other genetic abnormalities, environmental teratogens and some nutritional deficiencies account for some congenital anomalies but the majority are of unknown etiology. The hypothesis is here proposed that a significant proportion of congenital anomalies and cerebral palsy of unknown etiology are attributable to a monozygotic multiple conception with monochorionic placentation and that these anomalies, even in singletons, may be explained by early, unrecognized or unrecorded loss of one conceptus in a monochorionic monozygotic conception. The pathological mechanism is hemodynamic instability with episodes of acute feto-fetal transfusion that produce ischemic organ impairment in either or both twins. The resultant clinical abnormality will depend on range of severity (fetal death, infant death, congenital anomaly, normal infant), site or combination of sites (which organ[s] present[s] with the congenital anomaly) and timing (early, middle or late in gestation as shown by variation in brain pathology that is observed). PMID- 16354496 TI - Genetic influence on reproductive behavior in female rhesus macaques. PMID- 16354497 TI - Problems with using sum scores for estimating variance components: contamination and measurement noninvariance. AB - Twin studies of complex traits, such as behavior or psychiatric diagnoses, frequently involve univariate analysis of a sum score derived from multiple items. In this article, we show that absence of measurement invariance across zygosity can bias estimates of genetic and environmental components of variance. Specifically, if the item responses are considered as multiple indicators of a latent factor, and the aim is to partition the variance in the latent factor, then the factor loadings relating the items to the factor should be equal for monozygotic (MZ) and dizygotic (DZ) twins. While it seems unlikely, a priori, that these loadings should differ as a function of zygosity, certain special measurement situations are cause for concern. Ratings by parents, or self-ratings of phenotypes which are more easily observed in others than via introspection, may be tainted by the co-twin's phenotype to a greater extent in MZ than DZ pairs. We also show that the analysis of sum scores typically biases both MZ and DZ correlations compared to the true latent trait correlation. These two sources of bias are quantified for a range of values and are shown to be especially acute for sum scores based on binary items. Solutions to these problems include formal tests for measurement invariance across zygosity prior to analysis of the sum or scale scores, and multivariate genetic analysis at the individual item or symptom level. PMID- 16354498 TI - Parameterization of sex-limited autosomal linkage analysis for Mx. AB - Incorporation of sex-limitation (genotype-sex interaction) effects into a model of quantitative trait loci ( QTL) analysis has been shown to increase the power to detect linkage when analyzing traits in which sex limitation is present (Towne et al., 1997). The present note provides a parameterization of the nonscalar sex limitation ACE model incorporating autosomal sex-limited QTL effects for use with the Mx matrix algebra program (Neale et al., 2002). An example script designed for use with extended sibships that takes advantage of the versatile treatment of covariates within Mx is included. PMID- 16354499 TI - Limitations of DSM-IV operationalizations of alcohol abuse and dependence in a sample of Australian twins. AB - Alcohol abuse and dependence are among the most common psychiatric conditions identified in epidemiological surveys of the general population. The aim of this article is to examine the psychometric properties of Diagnostic and Statistical Manual of Mental Disorders, (4th ed.; DSM-IV; American Psychiatric Association, 1994) criteria for alcohol abuse and dependence using latent class analysis (LCA). Six thousand two hundred and sixty-five young Australian twins (median age 30 years) were interviewed by telephone between 1996 and 2000 using a modified version of the Semi-Structured Assessment for the Genetics of Alcoholism (SSAGA). DSM-IV symptoms of alcohol abuse and dependence were collected by structured diagnostic interview and analyzed using methods of LCA. LCA revealed a 4-class solution for women that classified individuals according to the severity of their alcohol-related problems: no/few problems (66.5%), heavy drinking (23.9%), moderate dependence (7.6%) and severe dependence (2.0%). Among men the preferred solution included 5 classes corresponding to no/few problems (46.4%), heavy drinking (34.3%), moderate dependence (12.2%), severe dependence (3.0%) and abuse (4.0%). Evidence of a male-specific class of alcohol-related problems corresponding to abuse partially supports the DSM conceptualization of alcohol use disorders but suggests that this conceptualization--and measurement--may need to be refined for women. Identification of a male-specific abuse class also has important implications for interventions and treatment as these individuals experienced significant alcohol-related problems and comprised approximately 21% of all men classified with an alcohol use disorder. PMID- 16354500 TI - The etiology of mathematical and reading (dis)ability covariation in a sample of Dutch twins. AB - The genetic etiology of mathematical and reading (dis)ability has been studied in a number of distinct samples, but the true nature of the relationship between the two remains unclear. Data from the Netherlands Twin Register was used to determine the etiology of the relationship between mathematical and reading (dis)ability in adolescent twins. Ratings of mathematical and reading problems were obtained from parents of over 1500 twin pairs. Results of bivariate structural equation modeling showed a genetic correlation around .60, which explained over 90% of the phenotypic correlation between mathematical and reading ability. The genetic model was the same for males and females. PMID- 16354501 TI - Genetic etiology of reading difficulties in boys and girls. AB - Although it has been suggested that genetic influences on reading difficulties may differ in boys and girls, results obtained from previous analyses of data from same-sex twin pairs have failed to provide evidence for a differential genetic etiology of reading disability (RD) as a function of gender. However, results of a recent study in which data from both same-sex and opposite-sex twin pairs were analyzed indicated a higher heritability for reading difficulties in boys (Harlaar et al., 2005). Because the current sample of twin pairs tested in the Colorado Learning Disabilities Research Center is substantially larger than that analyzed for our previous report (Wadsworth et al., 2000), this hypothesis was tested more rigorously using data from both same-sex and opposite-sex twin pairs in our current augmented sample. Composite reading scores from 634 twin pairs were subjected to DeFries-Fulker sex-limitation analysis using the model fitting approach of Purcell and Sham (2003). Analysis of data from the combined sample of male and female twins indicated that genetic influences account for more than half the proband reading deficit (h2g = .58). When this model was extended to test for gender differences in the magnitude of genetic influences on RD, h2g estimates were somewhat higher for females than for males (.63 and .53, respectively), but the difference was nonsignificant (p > .3). A test for qualitative gender differences was also nonsignificant. Thus, these results provide little evidence for a differential genetic etiology of RD in boys and girls. PMID- 16354502 TI - Multivariate genetic analysis of academic skills of the Queensland core skills test and IQ highlight the importance of genetic g. AB - This study examined the genetic and environmental relationships among 5 academic achievement skills of a standardized test of academic achievement, the Queensland Core Skills Test (QCST; Queensland Studies Authority, 2003a). QCST participants included 182 monozygotic pairs and 208 dizygotic pairs (mean 17 years +/- 0.4 standard deviation). IQ data were included in the analysis to correct for ascertainment bias. A genetic general factor explained virtually all genetic variance in the component academic skills scores, and accounted for 32% to 73% of their phenotypic variances. It also explained 56% and 42% of variation in Verbal IQ and Performance IQ respectively, suggesting that this factor is genetic g. Modest specific genetic effects were evident for achievement in mathematical problem solving and written expression. A single common factor adequately explained common environmental effects, which were also modest, and possibly due to assortative mating. The results suggest that general academic ability, derived from genetic influences and to a lesser extent common environmental influences, is the primary source of variation in component skills of the QCST. PMID- 16354503 TI - Familial clustering of major depression and anxiety disorders in Australian and Dutch twins and siblings. AB - The aim of this study was to investigate familial influences and their dependence on sex for panic disorder and/or agoraphobia, social phobia, generalized anxiety disorder and major depression. Data from Australian (N = 2287) and Dutch (N = 1185) twins and siblings who were selected for a linkage study and participated in clinical interviews to obtain lifetime Diagnostic and Statistical Manual of Mental Disorders (4th ed.; DSM-IV) diagnoses were used. In a liability model, tetrachoric correlations were estimated in sibling pairs and sex differences between sibling correlations were tested. For each diagnosis, the sibling correlations could be constrained to be equal across the Australian and Dutch samples. With the exception of panic disorder and/or agoraphobia, all sibling correlations were the same for brother, sister and opposite-sex sibling pairs and were around .20. For panic disorder and/or agoraphobia, the correlation was .23 in brother and sister pairs, but absent in opposite-sex sibling pairs. From these results it can be concluded that upper heritability estimates, based on twice the correlations in the sibling pairs, vary between 36% (major depression) and 50% (social phobia). Furthermore, different genetic risk factors appear to contribute to the vulnerability for panic disorder and/or agoraphobia in men and women. No other sex differences were found. PMID- 16354504 TI - Association between dopamine D4 receptor exon III polymorphism and emotional reactivity as a temperamental trait. AB - The association between high and low levels of emotional reactivity (ER) as a temperamental trait and the dopamine D4 receptor (DRD4) exon III polymorphism in healthy men aged 18 to 27 (M = 21.03, SD = 2.23) was examined. ER, measured by the Formal Characteristics of Behavior-Temperament Inventory (FCB-TI), is defined as a tendency to react intensively to emotion-generating stimuli and is expressed in high emotional sensitivity and in low emotional endurance. Data analysis demonstrated a statistically significant difference in the distribution of genotypic frequencies between the low and high ER groups: chi2 = 4.88; df = 1; p = .027, odds ratio (OR) =2.85, 95% confidence interval (CI) = 1.11-7.32. An insignificant difference in allele frequencies between the two groups was noted: chi(2) = 7.47; df = 3; p = .058; OR = 2.9, 95% CI = 1.29-6.53. These findings suggest a role of the DRD4 exon III polymorphism in the modulation of ER as a temperamental trait. Due to the preliminary nature of our findings, replication is necessary. PMID- 16354505 TI - A comparison of twin birthweight data from Australia, the Netherlands, the United States, Japan, and South Korea: are genetic and environmental variations in birthweight similar in Caucasians and East Asians? AB - Birthweight has implications for physical and mental health in later life. Using data from Caucasian twins collected in Australia, the Netherlands and the United States, and from East Asian twins collected in Japan and South Korea, we compared the total phenotypic, genetic and environmental variances of birthweight between Caucasians and East Asians. Model-fitting analyses yielded four major findings. First, for both males and females, the total phenotypic variances of birthweight were about 45% larger in Caucasians than in East Asians. The larger phenotypic variances were mainly attributable to a greater shared environmental variance of birthweight in Caucasians (ranging from 62% to 67% of variance) than Asians (48% to 53%). Second, the genetic variance of birthweight was equal in Caucasians and East Asians for both males and females, explaining a maximum of 17% of variance. Third, small variations in total phenotypic variances of birthweight within Caucasians and within East Asians were mainly due to differences in nonshared environmental variances. We speculate that maternal effects (both genetic and environmental) explain the large shared environmental variance in birthweight and may account for the differences in phenotypic variance in birthweight between Caucasians and East Asians. Recent molecular findings and specific environmental factors that are subsumed by maternal effects are discussed. PMID- 16354506 TI - Genetic and environmental influences on the handedness and footedness in Japanese twin children. AB - The purpose of this study was to examine the genetic contribution to handedness and footedness in childhood using one of the largest available databases of Japanese twins. The participants were 1131 twin pairs, 1057 males and 1205 females, of 11 or 12 years of age (6th grade of secondary school in the Japanese education system). All data were gathered by questionnaire. The prevalence of left (nonright) handedness was 15% in males and 13% in females. The prevalence of left (nonright) footedness was 13% in males and 11% in females. The similarities between twin pairs, estimated by concordance rates and tetrachoric correlations, suggested a slight genetic effect on male handedness, no genetic effect on female handedness, and no genetic effect on footedness in either sex. Structural equation modeling showed small genetic factors (11%) in male handedness and no genetic factors in female handedness. As to footedness, no genetic factors were observed in either sex. The effects of nonshared environmental factors were large (85%) in males and moderate (44%) in females. Moreover, handedness and footedness tended to be concordant irrespective of sex, with polychoric correlations over r = .70. The results of bivariate genetic analyses were not necessarily satisfactory. For males, no model fit. For females, shared and nonshared environmental factors explained the concordance of handedness and footedness. It was concluded that the genetic effects on handedness and footedness are relatively small, as is their association; moreover, considerably large twin samples are needed to obtain stable and appropriate results. PMID- 16354507 TI - Analysis of factors affecting birthweight, birth length and head circumference: study of Japanese triplets. AB - The aims of this study were to identify factors associated with birthweight, birth length and head circumference for triplets, and analyze these body size parameters at birth, especially head circumference, according to gestational age. The subjects of this study were 370 mothers and their 1109 triplet children (excluding one stillborn infant) who were born between 1978 and 2002. The gestational age proved to be the strongest contributing factor to birthweight, birth length and head circumference of the triplets. Moreover, sex was a significant factor affecting birthweight, birth length and head circumference. Male neonates had a higher birthweight, longer birth length and greater head circumference than female neonates. Birth order in triplets also had a significant effect on birthweight and head circumference. Lower birth-order neonates had a higher birthweight and greater head circumference. An effect of maternal pregravid body mass index (BMI) on both birthweight and birth length was observed. The birthweights of triplets born to women whose pregravid BMIs were more than 26.0 kg/m2 weighed an average of 150 g more than those of triplets born to women whose pregravid BMIs were less than 19.8 kg/m2, and the birth length of triplets born to women whose pregravid BMIs were more than 26.0 kg/m2 averaged 1.5 cm longer than those of triplets born to women whose pregravid BMIs were less than 19.8 kg/m2. Concerning head circumference, the median head circumference of male neonates was approximately 0.5 cm longer than female neonates. Compared to singleton neonates, the median head circumference of triplets was almost the same. PMID- 16354512 TI - Will we still be cutting in the 21st century? PMID- 16354513 TI - Enoxaparin for thromboprophylaxis in morbidly obese patients undergoing bariatric surgery: findings of the prophylaxis against VTE outcomes in bariatric surgery patients receiving enoxaparin (PROBE) study. AB - BACKGROUND: Obese patients undergoing bariatric surgery are at significant risk for venous thromboembolism (VTE). We performed a multicenter, retrospective survey to evaluate the safety and efficacy of enoxaparin for thromboprophylaxis in patients with morbid obesity undergoing primary bariatric surgery. METHODS: From January to December 2002, 668 patients who underwent primary bariatric surgery at 5 centers were analyzed retrospectively. Baseline patient demographics, objectively diagnosed cases of VTE, and bleeding events were recorded. Patients received enoxaparin preoperatively (30 mg) or postoperatively (40 mg) every 12 or 24 hours or upon discharge (30 mg every 24 hours for 10 days). RESULTS: Overall, there were 6 (0.9%) pulmonary embolisms (PE) and 1 (0.1%) occurrence of deep vein thrombosis (DVT); all but 1 occurred after the cessation of thromboprophylaxis. The highest incidence of VTE was at Center B, which did not administer perioperative thromboprophylaxis (1 DVT and 2 PEs). There were 6 (0.9%) severe bleeding complications: 3 at center D and 3 at center E. In Center B, 2 deaths were recorded (0.3%): 1 due to sepsis and 1 due to bleeding, with both occurring after thromboprophylaxis was discontinued. CONCLUSION: The administration of enoxaparin, in various dosing regimens, is safe for thromboprophylaxis in morbidly obese patients undergoing bariatric surgery. Fewer events occurred with perioperative prophylaxis initiated in the hospital. Because all thromboembolic events occurred after the cessation of thromboprophylaxis, extended thromboprophylaxis may be of value. PMID- 16354514 TI - Role of gastric acid in stomal ulcer after gastric bypass. AB - BACKGROUND: The pathogenetic mechanisms of stomal ulcer after Roux-en-Y gastric bypass (RYGBP) are unclear. In order to study the role of gastric acid, we measured acidity in the proximal pouch using a pH-sensitive probe. METHODS: 6 patients (5 females, mean age 45 years old at time of operation) with endoscopically confirmed stomal ulcer, were studied 2 to 6 years after RYGBP. All complained of epigastric pain that improved during proton pump inhibitor (PPI) therapy. Control subjects were 6 females (50 years old) who had had RYGBP at least 5 years earlier and denied symptoms of epigastric pain or heartburn. The pH sensitive probe (Digitrapper-pH, Medtronic) was passed through the nose to the proximal pouch, guided by the calculated distance and pH response. The probe was left in place for 4 hours. The percentage of time with pH <4 was calculated. RESULTS: The probe could be accurately positioned in the proximal pouch both in symptomatic patients and in controls as evidenced by the acid pH reaction. The proximal pouches of patients with stomal ulcer were significantly more exposed to acid compared to controls. The median percentage of time with pH <4 was 69% and 20% in the stomal ulcer and the control group, respectively (P<0.01). Barium follow-through excluded gastro-gastric fistula in stomal ulcer patients. CONCLUSION: RYGBP patients with stomal ulcer have increased acid production in their proximal pouch in comparison with asymptomatic RYGBP patients. Gastric acid appears to have an important role in the pathogenesis of stomal ulcer. PMID- 16354515 TI - Increased burst pressure in gastrointestinal staple-lines using reinforcement with a bioprosthetic material. AB - BACKGROUND: Leakage at an anastomosis is a major and often catastrophic complication of gastrointestinal (GI) surgery. Staple-line reinforcement with one of the several materials commercially available has been utilized to reduce the incidence of this complication. The bioprosthetic material, small intestinal submucosa (SIS, Surgisis((R)); Cook, Inc., Bloomington, IN) has found widespread applications in surgery. However, its ability to improve the durability of staple lines in GI surgery in terms of burst pressure has not been documented. We hypothesized that SIS reinforcement of staple-lines in healthy living GI tissue would increase durability, as determined by leak rates at increased intraluminal pressures, compared to unreinforced staple-lines. METHODS: Two healthy Yorkshire Cross pigs were subjected to midline laparotomy and underwent small intestinal division (n=28) with GIA stapling devices. Half of the staple-lines were reinforced with SIS. The staple-lines were then exposed to increased intraluminal pressures by means of a constant-rate dye solution infusion, until staple-lines exhibited visible leakage of the dye solution. The intraluminal pressure was recorded at the time of visible leakage. RESULTS: Staple-lines reinforced with SIS had significantly better durability as determined by analysis of variance and Kaplan-Meier survival calculations, with respect to leak rate as a function of intraluminal pressure (P<0.003). The mean burst pressure of the unreinforced staple-lines was 53 mmHg, while those staple-lines reinforced with SIS had a mean burst pressure of 83 mmHg. CONCLUSION: Reinforcement of stapled GI anastomoses with SIS significantly increases anastomotic burst pressure. These findings suggest a role for this material in GI surgery. PMID- 16354516 TI - Incidental finding of gastrointestinal stromal tumors (GISTs) during laparoscopic gastric bypass. AB - BACKGROUND: Gastrointestinal stromal tumors (GISTs) are rare tumors, accounting for <1% of all neoplasms of the alimentary tract. GISTs have not been previously reported in association with gastric bypass surgery. METHODS: This study is a retrospective review of 517 consecutive morbidly obese patients who underwent laparoscopic Roux-en-Y gastric bypass (LRYGBP) between January 2002 and August 2005. Incidental intraoperative findings of gastric GIST were recorded. RESULTS: 4 patients (0.8%) were noted to have GISTs intra-operatively upon inspection of the stomach prior to partition. All GISTs were identified along the anterior aspect of the upper third of the stomach and were removed by laparoscopic wedge excision with at least a 1 cm margin. The 4 tumors were <1 cm in size and all had immunohistochemical analysis positive for CD117 (c-kit). None of the tumors had determinants of malignant behavior (high mitotic rate, necrosis or pleomorphism). CONCLUSION: We have found a 0.8% incidence of gastric GISTs in our morbidly obese patients undergoing LRYGBP. All of these small, benign tumors were found incidentally in asymptomatic patients. This case series underscores the need to fully assess the stomach prior to gastric pouch formation. Without the ability to grossly determine the benign or malignant behavior of GISTs, all these tumors found incidentally should be resected with adequate margins. PMID- 16354517 TI - Effects of bariatric surgery on vitamin D status and secondary hyperparathyroidism: a prospective study. AB - BACKGROUND: Morbidly obese patients have been reported to present with vitamin D insufficiency and secondary hyperparathyroidism. Scattered data are available regarding the effects of bariatric surgery on vitamin D status. We studied calcium metabolism and vitamin D status before and after bariatric surgery. METHODS: In this prospective study, 64 patients (M5/F59) fulfilled the inclusion criteria (i.e. 2 calcidiol serum determinations in the winter season) among 457 morbidly obese individuals who underwent Roux-en-Y gastric bypass (RYGBP) a mean of 36 months previously. Laboratory data (serum calcium, phosphorus, creatinine, alkaline phosphatase, albumin, calcidiol, albumin and iPTH) were determined before and after RYGBP. Pre- and postoperative calcidiol levels were categorized as being normal (>50 nmol/L), insufficient (25-50 nmol/L), and deficient (<25 nmol/L). Pre- and postoperative mild secondary hyperparathyroidism was defined as iPTH >7.3 pmol/L with simultaneous normal values for creatinine, calcium and phosphorus. RESULTS: RYGBP produced a significant weight loss coupled with a simultaneous increase in calcidiol (+28%, P<0.0005) and decrements in total alkaline phosphatase (-53%, P<0.0005) and iPTH (-74%, P=0.001). Corrected serum calcium, phosphorus, and creatinine levels were indistinguishable before and after RYGBP. Additionally, 37.5% of the patients maintained their calcidiol category, while 42.2 % improved it and 20.3% lost one category. CONCLUSIONS: RYGBP does not completely correct pre-existing vitamin D deficient states with secondary hyperparathyroidism. Low calcidiol bioavailability and or insufficient sunlight exposure do probably persist after bariatric surgery. While randomized controlled studies are warranted, it seems advisable to support vitamin D supplementation as well as increasing sunlight exposure in the morbidly obese population. PMID- 16354518 TI - The effects of acute preoperative weight loss on laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Minimal acute pre-operative weight loss significantly reduces liver size and intra-abdominal adipose tissue. We hypothesize that these changes will reduce intra-operative complications and reduce the difficulty of laparoscopic Roux-en-Y gastric bypass (LRYGBP). METHODS: This is a retrospective chart review of consecutive patients who had undergone isolated LRYGBP between July 2003 and March 2005. All patients participated in our institution's medically supervised Weight Management Program before surgery. RESULTS: 48 patients (Weight Loss Group) had an average percent loss of excess weight (%EWL) of 4.6; whereas 47 patients (No Weight Loss Group) gained an average of 4.8% of excess weight over an average period of 2.4 and 3 months (P=0.09), respectively. There were no differences between the two groups in age, gender, ASA class, co-morbidities, or BMI at operation. The Weight Loss Group had less intra-operative blood loss (102 vs 72 ml, P=.03). The surgeon was also less likely to report an enlarged liver in the Weight Loss Group (P=.02). Finally, the operation was less likely to deviate from the standard LRYGBP when patients lost weight (P=.02). No differences were seen in operative time, length of hospital stay, wound infections, or major complications. CONCLUSION: Acute preoperative weight loss is associated with less intra-operative blood loss and reduces the need for intraoperative deviation from the standard LRYGBP. A larger series with a greater reduction in excess weight is necessary to determine the maximal benefits of acute preoperative weight loss. PMID- 16354519 TI - Silastic ring vertical gastric bypass: evolution of an open surgical technique, and review of 1,588 cases. AB - BACKGROUND: Silastic ring vertical gastric bypass (SRVGBP) has evolved from a stapled (SSRVGBP) to a transected (TSRVGBP), and finally to a transected pouch with jejunal interposition (TSRVGBP with J-I). The creation of the gastroenterostomy evolved from a hand-sewn to a stapled and finally to a combined stapled and hand-sewn anastomosis. The circumference of the ring was increased from 5.5 to 6.0 cm. We address the effect of these modifications on surgical outcome. METHOD: The records of 1,588 consecutive patients (mean BMI of 44.5) since 1990 who had a SRVGBP were indentified from a prospective data-base of all patients undergoing bariatric operations. 205 patients with a prior bariatric operation were excluded from the review, leaving 1,383 patients who had a primary SRVGBP. RESULTS: In the 193 SRVGBP patients, there was 1 gastric leak (0.5%) and 64 gastrogastric fistulas (33.2%). In the 165 TSRVGBP patients, there were 4 gastric leaks (2.4%) and 14 gastrogastric fistulas (8.5%). In the 1,025 patients with TSRVGBP with JI, there were 8 gastric leaks (0.8%) and no gastro-gastric fistulas. In the TSRVGBP with J-I, 367 patients had a hand-sewn, 16 a stapled, and 642 a combined stapled and hand-sewn anastomosis. Stricture rate was 3.8%, 31%, and 2.6% respectively. There were 7 ring migrations (0.7%), all in the totally hand-sewn group. Ring removal was necessary in 20 (5%) with a 5.5-cm and 4 (0.74%) with a 6.0-cm ring. CONCLUSION: TSRVGBP with J-I with a combined stapled and hand-sewn gastrojejunal anastomosis using a 6.0-cm ring decreased the incidence of complications, and is our current technique. PMID- 16354520 TI - Incidence of regurgitation after the banded gastric bypass. AB - BACKGROUND: Frequent regurgitation is a common complication following Roux-en-Y gastric bypass (RYGBP). This study investigated the risk of becoming a chronic regurgitator, by considering silicone ring size and lower esophageal sphincter (LES) function, and their relationship with weight loss. METHODS: 80 morbidly obese patients were randomly selected to undergo surgery using ring length of 62 mm (40 patients, group A) or 77 mm (40 patients, group B), with 6 months' postoperative follow-up. Preoperative esophageal manometry parameters were correlated with occurrence of chronic postoperative regurgitation. Patients were considered to present chronic regurgitation when this occurred on >10 days/month. RESULTS: The groups were homogeneous regarding age, gender, race, weight, BMI (47.8+/-6.1 vs 50.2+/-6.4 kg/m2) and obesity-related diseases. There were 15% more chronic regurgitators in group A than in group B. Chronic regurgitators in group A lost more weight than chronic regurgitators in group B (P=0.026) or non chronic regurgitators in group A (P=0.016). A greater proportion of chronic regurgitators had LES hypotonia (mean respiratory pressure <14 mmHg) than did non chronic regurgitators (P=0.008). Logistic regression demonstrated that the chance of being a chronic regurgitator in group A was 4.5 times greater than in group B (P=0.046), and that the chance of a chronic regurgitator having LES hypotonia was seven times greater than of having normal LES pressure (P=0.006). CONCLUSION: Silicone ring size and LES hypotonia are independent prognostic factors for chronic regurgitation following RYGBP. Ring size and chronic regurgitation contribute significantly towards weight loss during the first 6 postoperative months. PMID- 16354521 TI - Duodenal switch has no detrimental effects on hepatic function and improves hepatic steatohepatitis after 6 months. AB - BACKGROUND: Nonalcoholic steatohepatitis (NASH) is a common histological finding on liver biopsy in morbidly obese patients. The condition, although benign, can progress to cirrhosis and liver failure. We investigated the effect of the duodenal switch (DS) operation on hepatic function and architecture, specifically hepatic steatosis and NASH. METHODS: Between November 1999 and June 2004, 697 DS operations were performed for the treatment of morbid obesity. A Tru-Cut needle liver biopsy was routinely performed during the DS operation. Liver function tests were drawn preoperatively for AST and ALT, and again postoperatively at 6, 12 and 18 months and yearly thereafter. Repeat Tru-Cut liver biopsy was performed on all patients (n = 78) who underwent a second intra-abdominal operation for any indication > or = 6 months postoperatively. The pathologist evaluated the 2 sets of liver biopsies in a blinded fashion. The hepatic adipose tissue content and the degree of hepatitis were compared in these patients. RESULTS: A transient worsening of the AST (13% of the baseline value, P<.02) and ALT ( 130-160% of the baseline value, P<.0001) levels was found at 6 months after the DS operation. Normal levels were achieved by 12 months postoperatively. A progressive improvement of about 3 grades in severity of NASH and a 60% improvement in hepatic steatosis, were seen by 3 years after the DS operation. CONCLUSION: DS improves both hepatic steatosis and its resulting inflammation. No detrimental effects on hepatic function were noted after 6 months. PMID- 16354522 TI - Varying blood monocyte and dendritic cell responses after laparoscopic versus open gastric bypass surgery. AB - BACKGROUND: Laparoscopy may activate innate immunity less than conventional open surgery. This may be important in obese patients who have pre-existing low-grade inflammation. This study examined phenotypic changes in blood monocytes (Mcs) and dendritic cells (DCs) from patients undergoing laparoscopic (L) or open (O) Roux en-Y gastric bypass (RYGBP) surgery. METHODS: 8 patients (3 male) had blood drawn before and after RYGBP, and on postoperative day (POD) 1, 3, and 28. Mc and DC quantity, phenotype, and activation status were determined by flow cytometry. RESULTS: Mean BMI was 53 +/- 4 and 46 +/- 1, and length of stay was 6.3 +/- 3.2 and 3.5 +/- 0.6 days, in the O (n=4) versus L (n=4) groups, respectively. Postoperative WBC count was 16 +/- 1 x 10(3)/mm3 after O and 10 +/- 1 x 10(3)/mm3 after LRYGBP (P<0.001). This was due to a greater rise in neutrophils and decline in lymphocytes after ORYGBP (P<0.001). Total Mcs increased in both groups at POD 1, but the number of CD18+ Mcs was reduced after ORYGBP (P=0.04). Mc human leukocyte antigen (HLA)-DR expression was lower in CD16+ Mcs after ORYGBP, suggesting decreased capacity to present antigen (P=0.002). Postoperatively, total DCs decreased in both groups, but recovered (P=0.04). The proportion and activation of the tolerogenic DC2 phenotype was lower, whereas the percentage of the ldDC phenotype was higher, in the O group (P=0.006). CONCLUSION: RYGBP changes the quantity and phenotype of circulating blood Mcs and DCs. Although there were overall similarities in the overall response to gastric surgery between open and laparoscopic, there were some notable differences, including a greater reduction in HLA-DR expression and increased number of immature DCs in the ORYGBP group. The findings suggest that RYGBP may have varying immunologic consequences depending upon the surgical procedure employed. PMID- 16354523 TI - In vivo band manometry: a new access to band adjustment. AB - BACKGROUND: By application of a newly developed device for invasive pressure measurements, we have investigated band adjustments monitored by in vivo intraband pressures. With access to the port of the gastric banding device, pressures can be recorded inside the band system at rest and during bolus application with different adjustments of the band. METHODS: 25 patients (mean age 38.7, mean BMI 45.1, 80% women) had intraband pressure measurements at the first band adjustment 8.2 weeks (range 6 to 17) postoperatively. For this purpose, we adapted a pressure monitoring system with the TruWave disposable pressure transducer of Edwards. All patients underwent gastric banding using the Swedish adjustable gastric band (SAGB) by the pars flaccida technique. RESULTS: In vivo intraband pressures differ from ex vivo intraband pressures. With increasing fill volume in vivo measurements show increasingly higher pressures than ex vivo measurements. This difference can mainly be attributed to the influence of the enclosed tissue. The in vivo intraband pressures correlate with the amount of outflow obstruction. CONCLUSION: Intraband pressure measurement is an encouraging new access to gastric banding. It appears to be a feasible method to control band adjustment without need for x-ray studies in low pressure bands. We expect physiologically exact adjustments to achieve good weight loss and to prevent esophageal problems in the long term. PMID- 16354524 TI - The influence of surgically-induced weight loss on the knee joint. AB - BACKGROUND: The causal relationship between obesity and osteoarthritis (OA) of the knee is generally accepted. Weight loss has been shown to reduce the development of OA and improve the radiological parameters of existing disease. However, inducing weight reduction is difficult, and thus the number of patients studied has been small. We wished to determine the effects of surgically-induced weight loss on objective, radiological evidence of OA in the knee joint. METHODS: 64 consecutive patients that were referred to the Bariatric Surgical Unit were enrolled in the study. The only exclusion criterion was the prior diagnosis of OA. Knee pain alone did not exclude patients from the study. The study was performed in a prospective manner as a before-after trial. Radiographic data was evaluated by an independent radiologist not involved in the patient care or follow-up. Upright film of the knee was taken prior to surgery and 3 months following surgery. Minimal medial joint space width (JSW) was measured by a digital image computer. In addition, patients were clinically assessed using the American Knee Society Score (AKSS) at these times. RESULTS: 59 of 64 patients were available for followup. BMI decreased from 43.4 to 36.9 (P<0.01). The medial joint space increased from 4.6 mm to 5.25 mm (P<0.001). The AKSS improved from 78.5 points (perfect function = 100 points) to 90.69 points (P<0.01). CONCLUSION: Surgically-induced weight loss is an effective, rapid and dependable means of reversing the radiological signs of early changes associated with OA. PMID- 16354525 TI - Adjustable gastric banding as a revisional bariatric procedure after failed gastric bypass. AB - BACKGROUND: Inadequate weight loss after proximal gastric bypass presents a clinical challenge to bariatric surgeons. Pouch size, stoma size and limb length are the variables that can be surgically altered. Aside from conversion to distal bypass, which may have significant negative nutritional sequelae, revisional surgery for this group of patients has not often been reported. The addition of adjustable silicone gastric banding (ASGB) to Roux-en-Y gastric bypass (RYGBP) may be a useful revision strategy because it has potential safety benefits over other revisional approaches. MATERIALS AND METHODS: We report on 8 patients who presented with inadequate weight loss or significant weight regain after proximal gastric bypass. All patients underwent revision with the placement of an ASGB around the proximal gastric pouch. Bands were adjusted at 6 weeks postoperatively and beyond as needed. Complications and weight loss at the most recent follow-up visit were evaluated. RESULTS: Mean age and body mass index (BMI) at the time of revision were 39 +/- 9.9 years and 44.0 +/- 4.5 kg/m2 respectively. No patients were lost to follow-up, and they lost an average of 38.1 +/- 10.4% and 44.0 +/- 36.3% of excess weight and 49.1 +/- 20.9% and 52.0 +/- 46.0% of excess BMI in 12 and 24 months respectively. Patients lost an average of 62.0 +/- 20.5% of excess weight from the combined surgeries in 67 (48-84) months. The only complication was the development of a seroma overlying the area of the port adjustment in one patient. There have been no erosions or band slippages to date. CONCLUSIONS: These results indicate that the addition of an ASGB causes significant weight loss in patients with poor weight loss outcome after RYGBP. The fact that no anastomosis or change in absorption is required may make this an attractive revisional strategy. Long- term evaluation in a larger population is warranted. PMID- 16354526 TI - The early effects of weight loss surgery on regional adiposity. AB - BACKGROUND: Weight loss beyond 6 months following laparoscopic adjustable gastric banding (LAGB) is associated with a preferential mobilization of visceral adipose tissue and an improvement in insulin sensitivity in insulin resistant subjects. Because the rate of weight loss is greatest in the first 3 months after LAGB, we investigated the impact of LAGB on changes in regional lipid deposition and insulin sensitivity over this period. METHODS: 10 female obese non-diabetic subjects underwent magnetic resonance (MR) imaging and spectroscopy before and 12 weeks after LAGB (using the Swedish band), for the quantification of abdominal subcutaneous and visceral adipose tissue areas and intrahepatic lipid. Fasting blood free fatty acids were analyzed. Insulin sensitivity was monitored by fasting insulin and homeostasis model assessment (HOMA). RESULTS: Median weight loss 12 weeks after gastric banding was 9.5 kg [interquartile range (IQR): -16.5 to -6]. There were significant reductions in median abdominal subcutaneous (-20% [IQR: -24 to -13]) and visceral (-15% [IQR: -49 to -8]) adipose tissue depots as well as plasma free fatty acids (-34% [IQR: -79 to -8]). The amount of weight lost was directly proportional to the initial BMI (r=0.778; P=0.008). Visceral fat loss was proportional to initial visceral adiposity (r=0.80, P=0.01). There was no significant improvement in insulin sensitivity. CONCLUSION: Significant fat loss occurs 3 months after LAGB. The absence of a concurrent improvement in insulin sensitivity may reflect the relatively small reduction in visceral adipose tissue at this stage. Improvement in insulin sensitivity beyond 3 months after LAGB may be due to the continued loss of visceral adipose tissue. PMID- 16354527 TI - The relationship between BMI, plasma leptin, insulin and proinsulin before and after laparoscopic adjustable gastric banding. AB - BACKGROUND: Morbid obesity is associated with over-secretion of leptin and insulin, and predisposes to development of carbohydrate intolerance. In the current study, we explored the impact of BMI after laparoscopic adjustable gastric banding (LAGB) on leptin, insulin and proinsulin levels. METHODS: 23 obese patients (8 males, 15 females) were included in the study. Their mean age was 36+/-6 yrs (range 21-56 yrs). Blood samples were collected for measurement of plasma leptin, insulin and proinsulin before and 6 and 14 months after LAGB. RESULTS: Mean BMI before surgery was 46.04 +/- 4.44 kg/m2, with significant and equal reduction of 18% in each of the follow-up periods, with total BMI reduction of 33% (P <.0001). The levels of circulating leptin, insulin and proinsulin before intervention were 119.3 +/- 53.1 ng/ml, 159 +/- 13 pmol/l, and 36.36 +/- 23.06 pmol/l respectively. Despite an equal BMI reduction in the 2 follow-up periods, the most significant decrease in hormone levels was observed in the immediate postoperative period (54, 53, and 45%, respectively), when compared to the second follow-up period (15, 30, 10%, respectively). The highest total decline in hormone level of 70% was obtained with insulin, compared to 52% in leptin, and 50% in proinsulin. Despite the significant decrease in proinsulin and insulin levels, their ratio increased from 0.22, to 0.28 and 0.36 after LAGB. Unlike insulin and proinsulin, leptin levels strongly and persistently correlated with BMI during the study. CONCLUSION: Following LAGB, weight loss was associated with decreased levels of circulating leptin, insulin and proinsulin, most prominent in the first follow-up period. Unlike insulin and proinsulin, leptin showed the most significant and persistent correlation with BMI, suggesting that morbid obesity acts through different feedback hormonal mechanisms which are probably not regulated only by absolute weight loss. Longer follow-up and larger numbers of patients are needed to clarify long-term hormonal profile, as well as the beneficial lasting effects of such interventions. PMID- 16354528 TI - Alterations in T-cell subset frequency in peripheral blood in obesity. AB - BACKGROUND: Obesity affects the regulation of immune and inflammatory responses. This study characterizes differences in peripheral blood lymphocyte phenotype in obese humans. METHODS: Frequencies of lymphocyte subsets among peripheral blood mononuclear cells were compared between 10 obese (BMI > or = 35) and 10 lean subjects, as determined by antibodies directed against cluster differentiation (CD) markers. RESULTS: Obese patients demonstrated an increased frequency of CD3+CD4+ T-cells (mean difference 12%, P=0.004), a decreased frequency of CD3+CD8+ T-cells (mean difference 9.4%, P=0.016) and an increased frequency of CD3+CD8+CD95+ T-cells (mean difference 13.3%, P=0.032). No other differences among T-cell or monocyte subsets were noted. CONCLUSIONS: Obesity is associated with alterations in frequencies of peripheral CD4+ and CD8+ T-cells and aberrations in the expression of CD95 among CD8+ T-cells. These data suggest both CD4+ and CD8+ T-cell compartments, as well as the regulation of CD95 expression on CD8+ T-cells, as targets for further study into obesity's effects on the immune system. PMID- 16354529 TI - Results of laparoscopic sleeve gastrectomy (LSG) at 1 year in morbidly obese Korean patients. AB - BACKGROUND: In Asia, the type and main cause of obesity is different than in western society. Therefore, the treatment plan should be differentiated, and the surgery for morbid obesity should be carefully chosen. The early results of laparoscopic sleeve gastrectomy (LSG) without duodenal switch that has been performed in the Korean population is reported. METHODS: We retrospectively reviewed 130 patients who underwent LSG from January 2003 to May 2004. 60 of these patients now had >1 year of regular follow-up, and are the subject of this report. LSG was performed through 4 12-mm ports and 1 15-mm port, using the Endo GIA stapler to create a lesser curve gastric tube over a 48-Fr bougie. RESULTS: For the 60 patients, the postoperative EWL was 71.6+/-21.9% at 6 months and 83.3+/-28.3% at 12 months. At 12 months after LSG, decrease in BMI was 9.2+/-3.7 kg/m2, and median weight loss was 24.6+/-10.0 kg. Dyslipidemia resolved in 75% of patients within 12 months. Diabetes resolved in 100% of patients within 6 months of operation. Hypertension resolved in 92.9% and improved in 100%. Joint pain resolved in 100% within 12 months. Weight loss plateaued at 12 months in the majority of patients. One patient has undergone a malabsorption procedure (duodenal switch) as a second-stage operation. CONCLUSION: Additional studies and follow-up are needed to determine the best surgical treatment for morbidly obese Asian patients. However, LSG without the second-stage duodenal switch operation has been an effective weight loss operation thus far, in most of the Korean patients. PMID- 16354530 TI - Recommendations regarding obesity surgery. AB - In France, 1,000 obese persons per month undergo a bariatric operation. Obesity surgery requires coordination and monitoring of aftercare. The French public health-care insurer asked the medical associations involved in obesity management to provide guidelines for obesity surgery. The recommendations were developed by the national associations of Obesity, Nutrition and Diabetes: the Association Francaise d'Etudes et de Recherches sur l'Obesite (AFERO), member of the EASO and IASO; the Association de Langue Francaise pour l'Etude du Diabete et des Maladies Metaboliques (ALFEDIAM); the Societe Francaise de Nutrition (SFN); and the Societe Francaise de Chirurgie de l'Obesite (SOFCO). This article presents the short version of the guidelines. PMID- 16354531 TI - Laparoscopic Roux-en-Y gastric bypass is safe and feasible after orthotopic liver transplantation. AB - Liver transplantation has become a life-saving procedure for patients with end stage liver disease. Since obesity is rampant in our society, it is not surprising that patients with a liver transplant suffer from obesity as well. In addition, Roux-en-Y gastric bypass (RYGBP) is a life-saving procedure for patients suffering from morbid obesity. However, a liver transplant is an extensive and invasive abdominal procedure. Further operations after orthotopic liver transplantation may be challenging. With increasing surgeon experience and technical advancements in laparoscopic equipment, previous abdominal surgery is no longer a contraindication to utilize the laparoscopic approach for RYGBP. In fact, herein, we describe the first laparoscopic RYGBP after orthotopic liver transplantation in the world literature. PMID- 16354532 TI - Late gastric pouch necrosis after Lap-Band, treated by an individualized conservative approach. AB - Late band slippage has occurred in nearly 3-10% of patients after laparoscopic adjustable gastric banding (LAGB) with an average delay of 13 months. Band slippage can rarely lead to necrosis of the enlarged pouch, a potentially life threatening condition. We report a female (BMI 39.92 with co-morbidities) who developed acute outlet obstruction 2 years after LAGB placement. After prompt band deflation, an urgent Gastrografin swallow showed stomach slippage without emptying. At re-operation pouch strangulation was discovered. The pouch appeared to be ill-fated, but as no tear was evident on intra-operative assessment, we decided to simply remove the band and drain. The patient was successfully discharged after 8 days, and the last upper endoscopy showed only a large ulcer in the fundus that was healing. Proper and prompt management of symptomatic patients with stomach slippage, with early operation when acute obstruction is evident, can enable a successful stomach-sparing approach. PMID- 16354533 TI - Emergence of pica (ingestion of non-food substances) accompanying iron deficiency anemia after gastric bypass surgery. AB - Pica, the compulsive ingestion of non-food substances, is a common, yet less well known symptom of iron deficiency anemia (IDA). Ice eating or pagophagia is one of the most common forms of pica associated with iron deficiency and IDA, and has only recently been identified among patients who had undergone Roux-en-Y gastric bypass (RYGBP). This article presents additional cases of pagophagia in women which emerged after RYGBP and resolved with iron treatment. These cases, added to the literature, lead to further discussion of how healthcare providers can subtly probe patients for the presence of pica. PMID- 16354534 TI - Rhabdomyolysis following laparoscopic gastric bypass. AB - Rhabdomyolysis is a rare complication of serious surgical procedures, and constitutes a clinical and biochemical syndrome, caused by injury and destruction of skeletal muscles. It is accompanied by pain in the region of the referred muscle group, increase in creatine phosphokinase levels, myoglobinuria, often with severe renal failure, and finally multi-organ system failure and death, if not treated in time. The main risk factor in the development of postoperative rhabdomyolysis is prolonged intraoperative immobilization of the patient. Morbidly obese patients who undergo laparoscopic bariatric operations should be considered high-risk for rhabdomyolysis, from extended immobilization and pressure phenomena in the lumbar region and gluteal muscles. We report a 20-year old woman with BMI 51, who underwent a prolonged laparoscopic Roux-en-Y gastric bypass. Postoperatively, she presented severe myalgia in the gluteal muscles and lumbar region, oliguria and creatine phosphokinase levels that reached 38,700 U/L. She was treated with intensive hydration and analgesics, and did not develop acute renal failure because diagnosis and treatment were attained immediately. PMID- 16354535 TI - Ghrelin levels and sleeve gastrectomy in super-super-obesity. PMID- 16354536 TI - Gordon Shaw. PMID- 16354537 TI - Long-term enhancement of maze learning in mice via a generalized Mozart effect. AB - OBJECTIVES: An animal model of the 'generalized Mozart effect' (GME) - enhanced/normalized higher brain function in response to music exposure - has been established. We extend those results in two studies using another species (mice). Study 1: (1) maze testing after music exposure was extended to a minimum of 6 hours; (2) no exposure to music in utero. Study 2: (1) music exposure time further reduced; (2) maze testing extended to 24 hours. METHODS: Study 1: two mouse groups were exposed to music continuously for 10 hours per day for 10 weeks (Group I: Mozart's Sonata K.448, Group II: Beethoven's Fur Elise). After 10 weeks, the ability to negotiate a T-maze was assessed (recording working time in maze, number of errors). Maze ability was tested 6 hours following the last music exposure. Study 2: two mouse groups were exposed periodically to music (58% silence) 10 hours per day for 10 weeks. Experiments after 10 weeks examined the groups' abilities to run the maze (recording working time/errors). Experiments were conducted 24 hours following the last music exposure. RESULTS: The Mozart group exhibited significant enhancements compared with the control mice in both studies, i.e. significantly lower working time (p<0.05) and committed fewer errors. DISCUSSION: Observation of GME in another species supports its generality for the mammalian cortex. The absence of a GME in fMRI studies for the control music also indicates a neurophysiological basis. With extended exposure, GME is a long-term effect, indicating potential clinical importance. It has been demonstrated that GME reduces neuropathological spiking significantly in epileptics. We discuss the relevance of this study for epilepsy treatment. PMID- 16354538 TI - Dihydroxy-2,5 benzenesulphonate (dobesilate) elicits growth arrest and apoptosis in glioma cells. AB - OBJECTIVES: Dihydroxy-2,5 benzenesulphonate (dobesilate) is used as an oral agent for treatment of vascular complications of diabetic retinopathy. We previously showed that blockade of fibroblast growth factor (FGF) driving angiogenesis with dobesilate inhibited new blood vessel formation in a mouse gelatine plug assay. In the present study we assessed the effects of dobesilate in rat glioma cells. METHODS: Rat C6 cells line were grown as adherent cells in Dulbecco modified Eagle medium supplemented with 1% (v/v) fetal bovine serum and antibiotics. Calcium dobesilate was added in independent experiments at the following concentrations: 10, 25, 50, 75 and 100 microM, and cells were incubated for 24 hours. Effects of dobesilate in glioma cell proliferation and survival were assessed using crystal violet staining and TUNEL assay, respectively. RESULTS: Incubation of glioma cells with dobesilate for 24 hours concentration-dependently decreased cell proliferation with an apparent IC50 of 25 microM, and this antiproliferative effect was related to a significant increase in glioma cell apoptosis. CONCLUSIONS: The results suggest that dobesilate is a promising candidate leading to the development of a new adjuvant therapeutic strategy for gliomas. PMID- 16354539 TI - Differential expression of angiopoietin-1 and angiopoietin-2 may enhance recruitment of bone-marrow-derived endothelial precursor cells into brain tumors. AB - OBJECTIVES: Angiogenesis is necessary for sustained neoplastic development. The angiopoietins Ang-1 and Ang-2 have been implicated in the regulation of this process; recent reports have suggested that a net gain in Ang-2 activity may be an initiating factor for tumor angiogenesis. We examined the recruitment of bone marrow-derived endothelial precursor cells into developing tumor neovasculature, and the spatial relationship between these cells and angiopoietin (Ang-1 and Ang 2) expression. METHODS: For this study T-cell depleted knockout mice (RAG-2/KO 5.2) were lethally irradiated and their bone marrow was reconstituted by bone marrow cells (BMCs) from transgenic mice (C57BL/Ka-Thy1.1) expressing green fluorescent protein (GFP). Rat glioma cells (RT-2/RAG) were then injected into the transplanted animals to form solid brain tumors. The animals were killed and their brains were analysed using immunohistochemistry and fluorescence-activated cell sorting. RESULTS: We found that BMCs migrated preferentially into the tumor when compared to adjacent healthy brain parenchyma. Furthermore, GFP+/CD34+ cells represented up to 8% of endothelial-like cells within the walls of tumor blood vessels. In the tumor, significant colocalization of Ang-2 with GFP+/CD34+ cells was noted (>80%), but colocalization with Ang-1 never exceeded 20%. In normal tissue directly surrounding the tumor, GFP+/CD34+ cells colocalized strongly with both angiopoietins (>75% and >70% for Ang-1 and Ang-2, respectively). DISCUSSION: The relative increase in angiopoietin-2 activity in brain tumors may result in the creation of a pro-angiogenic environment that enhances the recruitment of putative bone marrow-derived endothelial precursor cells into the tumor's developing vascular tree. PMID- 16354540 TI - Angiogenic capacity of human omental stem cells. AB - OBJECTIVES: The goals of the present study are to obtain, expand and characterize a stem cell population from human omentum and to evaluate its in vivo angiogenic capacities. METHODS: Human omental CD34+ cells were obtained from samples of human omentum by density gradient centrifugation in Ficoll. Proliferative pattern, marker expression (by flow cytometry) and angiogenic growth factor synthesis by omental cell cultures were determined. In vivo angiogenic capacity of the cells was evaluated in rats. RESULTS: Omental stem cells showed a high rate of proliferation (Ki67 staining), expressed CD34 marker and synthesized bFGF and VEGF. When implanted in rats, omental cells promoted neovascularization. Human omental cells were localized in rat tissue, mainly forming the endothelium of neo-vessels. Implantation of omental cells also facilitated angiogenesis of rat origin. CONCLUSION: CD34+ cell population of human omentum could be responsible for the clinical benefit of omental transplantation by promoting angiogenesis and synthesizing angiogenic growth factors to facilitate revascularization of injured tissue. PMID- 16354541 TI - Transplantation of embryonic stem cell-derived neural stem cells for spinal cord injury in adult mice. AB - AIMS: To investigate the efficacy of embryonic stem cell-derived neural stem cells (NSCs) for spinal cord injury (SCI) in mice and whether a combination treatment with thyroid hormone provides a more effective ES cell-based therapy. METHODS: Nestin-positive NSCs were induced from undifferentiated mouse ES cells by a step-by-step culture and used as grafts. Thirty-six mice were subjected to an SCI at Th10 and divided into three groups of 12. Graft cells were transplanted into the injury site 10 days after injury. Group 1 mice were left under observation without receiving graft cells, while mice in Group 2 received 2 x 104 graft cells, and those in Group 3 received 2 x 104 graft cells and were treated with a continuous intraperitoneal injection of thyroxin using osmotic mini-pumps. Behavioral improvement was assessed by a scoring system throughout the experimental period until post-transplantation day (PD) 28. RESULTS: Mice in Groups 2 and 3 demonstrated an improved behavioral function, as compared to those in Group 1 after PD 14. There was no significant difference in behavioral recovery between Groups 2 and 3. CONCLUSIONS: Transplantation of ES-NSCs into the injury site was effective for SCI, while thyroxine did not deliver additional effectiveness. PMID- 16354542 TI - Targeted gene therapy to antigen-presenting cells in the central nervous system using hematopoietic stem cells. AB - BACKGROUND: Hematopoietic stem cells (HSC) have been previously used as vectors for gene therapy of systemic disease. The effectiveness of HSC-mediated gene therapy largely depends on efficient gene delivery into long-term repopulating progenitors and targeted transgene expression in an appropriate progeny of the transduced pluripotent HSCs. In the present study, we examined the feasibility of using HSC transduced with self-inactivating (SIN) lentiviral vectors for the delivery of gene therapy to the central nervous system (CNS). MATERIAL AND METHODS: We constructed two SIN lentiviral vectors, EF.GFP and DR.GFP, to express the green fluorescent protein (GFP) gene controlled solely by the promoter of either a housekeeping gene EF-1alpha or the human HLA-DRalpha gene, which is selectively expressed in antigen-presenting cells. RESULTS: We demonstrated that both vectors efficiently transduced human pluripotent CD34+ cells capable of engrafting NOD/SCID mice. Only the DR.GFP vector mediated transgene expression in the murine CNS containing human HLA-DR+ cells. These cells express surface markers characteristic of resident CNS microglia. Furthermore, human dendritic cells derived from transduced and engrafted human cells potently stimulated allogeneic T cell proliferation. CONCLUSIONS: The present study demonstrated successful targeting of transgene expression to CNS microglia after stable gene transduction of pluripotent HSC. PMID- 16354543 TI - Therapeutic efficacy of intraventricular cyclosporine A and methylprednisolone on a global cerebral ischemia model in rats. AB - OBJECTIVE: Cyclosporine A (CsA) with its immunosuppressive actions and methylprednisolone (MP) as a free radical scavenger were suggested together to alleviate neural tissue damage after an ischemic insult. The aim of this study was to investigate neuroprotective properties of CsA and MP in a global cerebral ischemia model. METHODS: Twenty-eight male Sprague-Dawley rats were divided randomly into four separate groups: CsA, MP, sham and control. Global cerebral ischemia was performed with the four-vessel occlusion model. After 30 minutes of ischemia, reperfusion was started with concomitant intraventricular administration of saline, MP (20 mg/kg) and CsA (10 mg/kg) into the lateral ventricle. Lipid peroxidation levels were measured from all experimental groups. Rats subjected to global cerebral ischemia exhibited a significant increase in cerebral lipid peroxide levels 6 hours after the onset of reperfusion. Both CsA and MP treatment significantly attenuated the degree of lipid peroxidation in cerebral tissues (p<0.05). Histopathological examinations of the CA1 sector of the hippocampus verified the neuroprotective properties of MP and CsA. CONCLUSIONS: The results suggested the neuroprotective properties of both agents, emphasizing more potent protection against ischemia by CsA. It was proposed that CsA could have exerted this effect with the blockage of mitochondrial permeability transition (MPT) pores, which are also critical if the necrotic and apoptotic cascades of the cell are considered. MP is judged to be neuroprotective, particularly in terms of its effects on lipid peroxidation. In conclusion, CsA and MP are ascertained to be neuroprotective agents as long as they cross the blood-brain barrier. PMID- 16354544 TI - Reducing unnecessary morbidity from percutaneous thermocoagulation in the treatment of trigeminal neuralgia--part C: a starting point for a somatotopic map of the human Gasserian ganglion. AB - OBJECTIVES: An original method and technique has been designed to reduce the significant morbidity associated with techniques currently used for percutaneous thermocoagulation in the treatment of trigeminal neuralgia. The current report deals with the mathematical and biostatistical analysis of verbal responses gathered using such a method in an attempt, as a starting point, to establish the somatotopic organization of the human gasserian ganglion. METHOD: A correspondence analysis was used to validate verbal responses. These were ordered in three 34 x 34 matrices, according the initial sequence of 34 subsegments of the face, which was based on the operative experience of one of the authors. After using a filter for the consistency of responses, and a maximum threshold below 0.5 V, 967 responses from 99 patients were selected for analysis. The frequencies obtained from each subsegment were compared, using all the possible pairwise combinations of the subsegments of the face, and the sequences were ordered using the least contradictory criterion. RESULTS: The incidence of each verbal response within each trigeminal division was analysed, resulting in a proposal of a sequence of 20 subsegments of the gasserian ganglion, listed from the depth to the surface. DISCUSSION: From the strict clinical point of view, the somatotopic map of each individual is invariant and easily analysed over long time periods. Its precise knowledge is critical for inducing smaller, properly placed lesions, in order to avoid unnecessary morbidity from percutaneous thermocoagulation in the treatment of trigeminal neuralgia. The proposed sequence of the gasserian somatotopic organization will be hopefully a useful guide for those interested in trigeminal physiological organization as well as for the therapeutic exploration of gasserian trigeminal fibers. PMID- 16354545 TI - Impaired regeneration of bcl-2-lacking peripheral nerves. AB - OBJECT: The outcome of peripheral nerve damage in still not satisfactory, despite the general capacity of peripheral nervous system to regenerate. The molecular mechanisms underlying nerve regeneration are still not clear, but it is likely that apoptosis regulating genes plays a crucial role in these processes. The aim of the present study was to establish the role of the anti-apoptotic gene bcl-2 in peripheral nerve repair. MATERIAL AND METHODS: Sciatic nerves of bcl-2 deficient and wild type mice were transected, and immediately re-sutured. The regeneration was assessed functionally and morphologically throughout the 4-week follow-up. RESULTS: We found markedly worse sciatic function index outcome, as well as more significant atrophy of denervated muscles in bcl-2 knock-out animals when compared with wild-type ones. The intensity of histological regeneration features, including GAP-43-positive growth cones, Schwann cells and macrophages in the distal stump of the transected nerve, was also decreased. The number of motor and sensory neurons in the relevant cross-sections of spinal cord was similar in both groups of mice. CONCLUSION: We concluded that the bcl-2 gene plays an important role in peripheral nerve regeneration, influencing nerve injury site clearing, fiber regrowth and myelination. PMID- 16354546 TI - Carotid endarterectomy without shunt: the role of cerebral metabolic protection. AB - BACKGROUND: The optimal method to protect the brain from hemodynamic ischemia during carotid endarterectomy (CEA) remains controversial. This study reports our experience with induced arterial hypertension and selective etomidate cerebral protection in a cohort of patients who underwent CEA without shunting and continuous electroencephalography (EEG) monitoring. METHODS: We reviewed retrospectively 102 consecutive CEAs performed in 102 patients with routine EEG monitoring and general anesthesia between March 1998 and October 2002. There were 65 (66%) symptomatic and 37 (34%) asymptomatic individuals. A protocol of induced arterial hypertension against EEG ischemic changes during carotid artery cross clamping was followed. Only patients with EEG changes refractory to induced hypertension went into etomidate-induced burst suppression. RESULTS: EEG changes were classified as mild, moderate and severe. Twenty patients (19.6%) developed asymmetric EEG changes, of which the great majority were mild and moderate (75%, p< 0.05). Seven patients with moderate (n=3) and severe (n=4) EEG changes needed etomidate cerebral protection. There were no mortalities and only one stroke (0.98%) is reported in the series. The morbidity rate was 6.8% and included transient cranial nerve palsies (n=5) and wound hematoma (n=1). CONCLUSIONS: Carotid endarterectomy can be safely performed with EEG monitoring and selective induced arterial hypertension and etomidate cerebral protection. Our results suggest that this method may be a good alternative for shunting and its inherent risks. PMID- 16354547 TI - Experimental cerebral arteriovenous fistulas. AB - OBJECTIVE: The aim of this paper is to analyse and summarize the main advances in experimental research on cerebral arteriovenous fistulas. METHODS: A detailed analysis of the literature and my own research experience were employed to outline the methodology whereby experimental cerebral arteriovenous fistulas are created and further studied. RESULTS: The analysis and quantification of the anatomical and functional variables in different experimental cerebral arteriovenous fistula models make it possible to develop more appropriate and individual ways of treatment in affected patients. CONCLUSION: Experimental research on cerebral arteriovenous fistulas helps physicians to understand and predict more accurately the future evolution of arteriovenous malformations in humans. PMID- 16354548 TI - Effects of VEGF and nitric oxide synthase inhibition on blood-brain barrier disruption in the ischemic and non-ischemic cerebral cortex. AB - OBJECTIVES: This study was performed to compare the effects of exogenous vascular endothelial growth factor (VEGF) and nitric oxide synthase (NOS) inhibition on blood-brain barrier (BBB) disruption in the ischemic cortex (IC) and non-ischemic contralateral cortex (CC) during the early stage of focal cerebral ischemia in rats. METHODS: A middle cerebral artery (MCA) was occluded after a craniotomy in each rat under isoflurane anesthesia. Two more craniotomies were performed over the contralateral non-ischemic hemisphere to expose cerebral cortex. For the control rats, the normal saline patches were applied to all three craniotomy holes (control group). To inhibit NOS, NG-nitro-L-arginine-methyl ester (L-NAME) (10 mg/kg) was administered i.v. 20 minutes after MCA occlusion (L-NAME group). In another group, VEGF (10(-10) M) was topically applied 30 minutes after MCA occlusion on the IC as well as one of the holes of the contralateral cortex (VEGF group). To investigate the effects of the combination of VEGF and L-NAME, both L NAME and VEGF were administered as described above (L-NAME+ VEGF group). The transfer coefficient (Ki) of 14C-alpha-aminoisobutyric acid and the volume of 3H dextran (70 000 Da) distribution were determined to measure the degree of BBB disruption at 1 hour after MCA occlusion. RESULTS: In the control group, Ki of the IC was significantly higher than the contralateral cortex (CC) (p<0.005). VEGF application increased the Ki of the IC further when compared with the control group (+51%, p<0.05%). L-NAME administration produced no significant decrease in the Ki of the IC when compared with the control group. With L-NAME+ VEGF administration, the Ki of the IC became significantly lower than that of the VEGF alone (-38%, p<0.005). Thus, L-NAME produced a much greater decrease in the Ki of the IC in the VEGF treated than the control animals (p<0.05). In the non IC, VEGF, L-NAME, or their combination did not affect BBB disruption. The volume of dextran distribution followed a similar pattern to Ki. DISCUSSION: Our data suggest that even in the early stage of focal cerebral ischemia, the degree of BBB disruption in response to the exogenous VEGF is much greater in the ischemic than in the non-IC and that the mechanism of the increase of BBB disruption by VEGF in the IC involves the NOS pathway. PMID- 16354549 TI - Physicochemical characteristics of soluble oligomeric Abeta and their pathologic role in Alzheimer's disease. AB - Extracellular fibrillar amyloid deposits are prominent and universal Alzheimer's disease (AD) features, but senile plaque abundance does not always correlate directly with the degree of dementia exhibited by AD patients. The mechanism(s) and dynamics of Abeta fibril genesis and deposition remain obscure. Enhanced Abeta synthesis rates coupled with decreased degradative enzyme production and accumulating physical modifications that dampen proteolysis may all enhance amyloid deposit formation. Amyloid accumulation may indirectly exert the greatest pathologic effect on the brain vasculature by destroying smooth muscle cells and creating a cascade of negative impacts on cerebral blood flow. The most visible manifestation of amyloid dis-equilibrium could actually be a defense mechanism employed to avoid serious vascular wall degradation while the major toxic effects to the gray and white matter neurons are mediated by soluble oligomeric Abeta peptides with high beta-sheet content. The recognition that dynamic soluble oligomeric Abeta pools exist in AD and are correlated to disease severity led to neurotoxicity and physical conformation studies. It is now recognized that the most basic soluble Abeta peptides are stable dimers with hydrophobic regions sequestered from the aqueous environment and are capable of higher order aggregations. Time course experiments employing a modified ELISA method able to detect Abeta oligomers revealed dynamic intermolecular interactions and additional experiments physically confirmed the presence of stable amyloid multimers. Amyloid peptides that are rich in beta-sheet structure are capable of creating toxic membrane ion channels and a capacity to self-assemble as annular structures was confirmed in vitro using atomic force microscopy. Biochemical studies have established that soluble Abeta peptides perturb metabolic processes, provoke release of deleterious reactive compounds, reduce blood flow, induce mitochondrial apoptotic toxicity and inhibit angiogenesis. While there is no question that gross amyloid deposition does contribute to AD pathology, the destructive potential now associated with soluble Abeta suggests that treatment strategies that target these molecules may be efficacious in preventing some of the devastating effects of AD. PMID- 16354550 TI - Cognitive impairment and mortality in older healthy Mexican subjects: a population-based 10-year follow-up study. AB - OBJECTIVES: To estimate the incidence of cognitive impairment (CI) among cognitively healthy, Mexican subjects, and to evaluate the impact of demographic and vascular factors on the conversion to CI and mortality. METHODS: 734 eligible subjects (aged 55 to >90 years) from a population-based sample were examined. The cognitive function of participants was assessed using the Mini-Mental State Examination (MMSE) every 2 years. The subjects were followed for an average of 3.2 years. The CI was defined using two sets of criteria: (i) moderate CI, as a drop to 25-21 on the MMSE at 2-year follow-up or a decrease of at least four points and (ii) severe CI, defined as a drop of 21 or less in MMES at follow-up. The incidence density and period prevalence were determined as epidemiological measures as well as the cumulative incidence as a risk measure. Kaplan-Meier survival curves were used to analyse the main points of interest: CI, dementia and mortality. RESULTS: The period prevalence of moderate CI was 20%, and 10% for severe CI. During 1959 person-years of follow-up, severe CI developed in 33 of the 361 participants. While during 2096 person-years of follow-up; 80 of 361 participants developed moderate CI. The rate of progression to severe CI in moderate CI subjects gradually increases with follow-up. Both, moderate and severe CI were associated with low educational level, higher age and higher mortality. CONCLUSIONS: Elderly people with moderate CI have an increased risk of severe CI. Moderate and severe CI are both predictive of higher mortality in Mexican subjects. PMID- 16354551 TI - Growth of rat cortical neurons on DuraGen, a collagen-based dural graft matrix. AB - OBJECTIVES: DuraGen, a collagen-based dural graft matrix, is frequently used in clinical neurosurgery. In the present study we examined whether DuraGen influenced neuron survival of or process growth from cerebral cortex neurons in culture. METHODS: Dissociated E19 rat cerebral cortical neurons were cultured at low density on poly-L-lysine or on cryostat-sectioned DuraGen. Neuron survival was assessed using morphological criteria, fluorescein diacetate (FDA) and propidium iodide (PI), nuclear staining and TUNEL labeling. Process growth was analysed using specific antibodies against MAP2 and the 200 kDa neurofilament subunit (NF-H) to identify dendrites and axons, respectively. RESULTS: In immature cultures (3 days in vitro, DIV), nearly 70% of the neurons remained viable in control and DuraGen-exposed cells. In mature cultures (10 DIV), approximately 45% of the neurons were viable. Survival was similar in DuraGen cultures and controls. Cell viability also was similar when DuraGen conditioned the medium, but was not in contact with the neurons. When 10-day-old cultures were treated with glutamate (100 mumol/l for 24 hours) to elicit excitotoxic injury, a 40% decrease in neuron survival was observed. DuraGen's presence neither exacerbated nor attenuated glutamate-induced excitotoxic neuron death. The amount of necrotic or apoptotic cells also was similar in control and DuraGen cultures. Finally, DuraGen had an equal ability to support both axon and dendrite growth as poly-L-lysine. CONCLUSION: Our findings demonstrate that DuraGen has no adverse effect on survival of or process growth from cerebral cortical neurons in vitro. These data support DuraGen's biosafety as a dural substitute in clinical neurosurgery. PMID- 16354552 TI - An editorial overview: HIV/AIDS in China. PMID- 16354553 TI - Commentary remarks on "HIV/AIDS in China". PMID- 16354554 TI - Commentary remarks on "HIV/AIDS in China". PMID- 16354555 TI - The HIV epidemic in China: history, response, and challenge. AB - The first case of AIDS was reported in 1985 in China, but by the early 21st century, the government estimated that there were 840,000 citizens living with HIV/AIDS. The number is increasing rapidly. The major risk groups are injection drug users (IDUSs; 43%) and former plasma donors (27%), but rates among heterosexual groups are rising rapidly. Sentinel surveillance was initiated in 1986, and now includes IDUs, men-who-have-sex-with-men, sexually transmitted disease clinic attendees, antenatal women, long-distance truck drivers, and sex workers. Although the government was slow to respond to the epidemic in the late 20th century, it has made a vigorous response in the early 21st century. Components of that response include implementation and evaluation of harm reduction programs for IDUs, education to increase knowledge and reduce stigma, treatment and social support for rural and poor HIV/AIDS patients, widespread testing, and increased funding for HIV/AIDS programs. International agencies have been generous in their support of the government initiatives. To successfully combat the epidemic, China needs to develop and train the necessary infrastructure to implement its intervention programs, particularly in the rural areas, to vigorously combat stigma and discrimination, support research especially in the universities and research institutions other than the China Centers for Disease Control, develop a system for efficient exchange of research and program information, and update legislation to reflect the current situation. PMID- 16354556 TI - Global human genetics of HIV-1 infection and China. AB - Genetic polymorphisms in human genes can influence the risk for HIV-1 infection and disease progression, although the reported effects of these alleles have been inconsistent. This review highlights the recent discoveries on global and Chinese genetic polymorphisms and their association with HIV-1 transmission and disease progression. PMID- 16354557 TI - Prevalence and evolution of drug resistance HIV-1 variants in Henan, China. AB - To understand the prevalence and evolution of drug resistant HIV strains in Henan China after the implementation of free antiretroviral therapy for AIDS patients. 45 drug naive AIDS patients, 118 AIDS patients who received three months antiretroviral therapy and 124 AIDS patients who received six months antiretroviral treatment were recruited in the southern part of Henan province. Information on general condition, antiretroviral medicines, adherence and clinical syndromes were collected by face to face interview. Meanwhile, 14 ml EDTA anticoagulant blood was drawn. CD4/CD8 T cell count, viral load and genotypic drug resistance were tested. The rates of clinical improvement were 55.1% and 50.8% respectively three months and six months after antiretroviral therapy. The mean CD4 cell count after antiretroviral therapy was significantly higher than in drug naive patients. The prevalence rate of drug resistant HIV strains were 13.9%, 45.4% and 62.7% in drug naive patients, three month treatment patients and six month treatment patients, respectively. The number of resistance mutation codons and the frequency of mutations increased significantly with continued antiretroviral therapy. The mutation sites were primarily at the 103, 106 and 215 codons in the three-month treatment group and they increased to 15 codon mutations in the six-month treatment group. From this result, the evolution of drug resistant strains was inferred to begin with the high level NNRTI resistant strain, and then develop low level resistant strains to NRTIs. The HIV strains with high level resistance to NVP and low level resistance to AZT and DDI were highly prevalent because of the AZT+DDI+NVP combination therapy. These HIV strains were also cross resistant to DLV, EFV, DDC and D4T. Poor adherence to therapy was believed to be the main reason for the emergence and prevalence of drug resistant HIV strains. The prevalence of drug resistant HIV strains was increased with the continuation of antiretroviral therapy in the southern part of Henan province. Measures, that could promote high level adherence, provide new drugs and change ART regimens in failing patients, should be implemented as soon as possible. PMID- 16354558 TI - Commentary on "Prevalence and evolution of drug resistance HIV-1 variants in Henan, China". PMID- 16354559 TI - High risk populations and HIV-1 infection in China. AB - China is currently experiencing one of the most rapidly expanding HIV epidemics in the world. Although the overall prevalence rate is still low, with a population of 1.3 billion, high-risk factors which have contributed to the HIV/AIDS epidemics worldwide continue to prevail in China, including a high rate of injecting drug use and needle sharing, commercial sex with low rates of condom use, and concurrent sex with both commercial sex workers and non-commercial casual or steady sex partners. In addition, there are increasing "double risk" populations overlapping drug users and sex workers, as well as increasing rates of STDs and HIV among high-risk populations. Sexual transmission, therefore, may serve as a bridge connecting high-risk populations with general populations. There is an urgent need to prevent the spread of HIV from these high-risk populations into the general population of China. PMID- 16354560 TI - MSM and HIV/AIDS in China. AB - This article profiles current status of spread and control of HIV/AIDS in China. China has a significant population of MSM (men who have sex with men) and they have been becoming very much alive in many ways since 1990s due to recent social changes. Some surveys indicate that great many of MSM are engaged in high-risk behaviors. In addition, majority of MSM have also experienced sexual encounters with women sometimes in their lives, which possibly contribute to spread of HIV to women. Some reports documented that HIV is becoming rampant among MSM since more than 1% of them are now infected. Political, cultural and custom elements could hinder intervention activities against HIV spread among MSM. Fortunately, many cities in China have seen that MSM were in cooperation with responsible institutions carrying out certain intervention measures. The general situation is promising. The authors forecast that the fast HIV spread among MSM of China could possibly be halted within several years when the authorities become more sensible to this issue, health service institutions offer unswerving efforts toward the MSM community and those who involve in MSM undertakes necessary responsibilities. PMID- 16354561 TI - Injection drug use and HIV/AIDS transmission in China. AB - After nearly three decades of being virtually drug free, use of heroin and other illicit drugs has re-emerged in China as a major public health problem. One result is that drug abuse, particularly heroin injection, has come to play a predominant role in fueling China's AIDS epidemic. The first outbreak of HIV among China's IDUs was reported in the border area of Yunnan province between China and Myanmar where drug trafficking is heavy. Since then drug-related HIV has spread to all 31 provinces, autonomous regions and municipalities. This paper provides an overview to HIV/AIDS transmission through injection drug use in China. It begins with a brief history of the illicit drug trade in China, followed by a discussion of the emergence of drug related AIDS, and a profile of drug users and their sexual partners who have contracted the virus or who are vulnerable to infection. It ends by summarizing three national strategies being used by China to address both drug use and AIDS as major health threats. PMID- 16354562 TI - Update on the laboratory diagnosis and monitoring of HIV infection. AB - In China, the estimated number of HIV infected cases is approaching one million. Although public education has been initiated for awareness and behavioral modification for this devastating infection, better diagnostic methods are needed to identify infected persons and manage infection. Simple and more accurate diagnostic tools have become available, particularly for early detection and to monitor treatment in those who receive anti-retroviral treatment. In this short review, we summarize some of the common and new methodologies that can be used in clinical laboratories, in the field, or in private laboratories. These range from simple antibody tests to more sophistical methods that are used to monitor disease progression and identify drug resistance. These tools can assist physicians, medical practitioners, and laboratory personnel to select suitable diagnostic tools for the diagnosis, blood screening, monitoring of disease progression, and for detection of drug resistance to anti-retroviral therapies. PMID- 16354563 TI - Current progress of China's free ART program. AB - China's Free ART Program was initiated in 2002 as an emergency response to save and improve the lives of AIDS patients living mainly in impoverished rural regions of central China. With little experience in HIV/AIDS treatment and care and resource limitations, China's efforts to provide widespread access to free antiretroviral therapy has been a process fraught with difficulty. However, the Free ART Program is progressing from an emergency response to a standardized treatment and care system. The development of national guidelines, training programs, a laboratory support network, a national patient database, programs for special populations such as children and patients living with co-infections, and operational research has improved the scope and quality of the free treatment program. As of June 30, 2005, a total of 19,456 patients in 28 provinces, autonomous regions, and special municipalities had received free ART. Challenges stemming from the nature of China's health system and patient population persist, but with strong government support and a diverse set of resources, China has the capacity to overcome these challenges and to provide nationwide access to high quality treatment and care. PMID- 16354564 TI - Care of HIV-infected patients in China. AB - Compared with high infection areas of the world, the total HIV infection rate in China is relatively low. Nonetheless, because of China's vast territory and large population, the potential infection risk must be taken seriously. In the next few years, needle sharing among injection drug users will remain the most common route of transmission for the HIV/AIDS epidemic in China. Unprotected sex is gradually becoming a major route of transmission. China began to implement HAART in 1999 according to international standards. Prior to 2003, there were only about 150 HIV/AIDS patients were treated with HAART in some clinical trials and about 100 HIV/AIDS patients were treated by private sources. Results of those treatments are the scientific basis for development of the therapeutic strategies in China. In March of 2003, the Chinese government initiated China CARES program. In November of 2003, the Chinese Ministry of Health announced a national policy of free ARV treatment to all HIV+ Chinese citizens who were in poverty and required ARV therapy. There are total of 19,456 HIV/AIDS patients received free ARV drugs to date in 159 regions and 441 towns. Current challenges are how to follow-up and evaluate those patients in the clinical settings. The longer the therapy is postponed, the more side effects and the higher probability of drug resistance are going to occur. It remains unclear, therefore, when HAART regimen should be started in the HIV/AIDS population in China. PMID- 16354565 TI - Effects of drug relief hospital-based AIDS educational methods on drug users. AB - The objective of this study is to explore a potentially effective training method for the hospital professionals to educate drug users and to enhance their knowledge of HIV infection. One hundred and sixty one subjects, who came from 13 different provinces and were admitted in a drug relief hospital in Beijing, were recruited for this study. The average age of these subjects was 35.21 +/- 6.24 year old. The average numbers of years for drug addiction were 7 years, and the average numbers of drug relief treatment received in the past was 5.5 times. The level of AIDS knowledge of these subjects, including pathogenic factors, source of infection, route of transmission and preventive measures, were evaluated before and after receiving the AIDS educational training to these drug users. Our results showed that there was a statistically significant increase (P<0.01) in the knowledge of HIV infection and prevention among these subjects. Positive attitude and behavioral tendencies toward HIV prevention were also improved. Therefore, it is imperative for the medical professionals to incorporate AIDS education into drug relief treatment to achieve the maximum effect on the knowledge of AIDS and improvement of positive attitudes and behaviors toward HIV prevention among drug users. PMID- 16354566 TI - Approaches to antiretroviral therapy in China. AB - China has recognized the threat of HIV to its population and responded with a national antiretroviral treatment (ART) program. However, high ART failure rates and the spread of resistance within populations are important realities to consider when developing and managing ART programs in China and worldwide. Concepts which will define treatment success and local and national programmatic goals are 1) access to ART, 2) durability of ART at the patient level, 3) scalability of treatment modalities, and the 4) sustainability of the program at the community or national level. In the face of limited resources, China must also consider when to start ARV therapy, which agents to use, when to switch them, and how to treat highly experienced patients with drug resistance. The optimal ARV regimen to start with is changing frequently with the introduction of new agents and the presentation of new data. Currently, a regimen including tenofovir, emtricitabine or lamivudine and a nonnucleoside reverse transcriptase inhibitor appears to have optimal characteristics to treat HIV/AIDS in China. However, critical to all of these choices is the evaluation of programs implemented to insure wide scale success. China has wisely begun this process of evaluating the performance of local programs through systematic monitoring and evaluation of treatment outcomes. This will allow regimens and programs that work to be expanded, and programs with high failure rates to be eliminated. In the end, evidence based data supporting treatment strategies will allow China to successfully confront its AIDS epidemic early and prevent its tragic consequences. PMID- 16354567 TI - Governmental policies on HIV infection in China. AB - This article is a general review of the evolvement of HIV/AIDS-related public policies in China since 1980's. It tracks the important laws, regulations and other governmental documents in regard to HIV/AIDS prevention mainly at central level. PMID- 16354568 TI - United Nations system efforts to support the response to AIDS in China. AB - In the last two years, we have seen a remarkable intensification in the response to AIDS in China. A number of organizations have joined and contributed to the efforts of the Chinese government in responding the AIDS epidemic in China. This article specifically describes the role of the United Nations (UN) in supporting and strengthening those responses. Achievements of the United Nations highlighted in the article include: strengthened leadership and political commitment to respond to AIDS; improved HIV/AIDS surveillance and information; expanded prevention efforts; improved treatment, care and support to people living with HIV and increased resources for AIDS programs. Additional roles of the United Nations system in the near future include strengthening national leadership by supporting the 'three ones', i.e., one national plan on AIDS; one national coordinating authority for AIDS; and one monitoring and evaluation system for AIDS. In addition, the UN system is expected to strengthen alignment and harmonization of activities of all international organizations and improved accountability and oversight. Remaining challenges identified include increasing awareness of AIDS and reducing stigma and discrimination; reducing vulnerability and risk behaviour among specific groups; providing improved treatment, care and support for people living with HIV; promoting stronger engagement by civil society, and; addressing the gender dimensions of AIDS. PMID- 16354569 TI - Chinese NGOs in action against HIV/AIDS. AB - Chinese non-governmental organizations (NGOs) have played a significant role in the battle against AIDS in the People's Republic of China. This article provides a brief overview of the structure of these organizations, as well as an analysis of their principle accomplishments. Of great significance in this analysis is the fact that Chinese NGOs have effectively dealt with many sensitive health education areas that government authorities have felt reluctant to handle directly. As such, they have provided an indispensable component in the HIV/AIDS prevention and control calculus on the mainland. PMID- 16354570 TI - International non-governmental actors in HIV/AIDS prevention in China. AB - International non-governmental organizations were among the first international actors that responded to the emergence of AIDS crisis in China. Since 1994, the number of international non-governmental organizations and charitable foundations working in AIDS related issue areas in China has grown steadily and substantially. Despite their organizational differences, most of these non governmental actors present the characteristics of independent mission, localized practice and diverse working focus. Even though they are constrained by financial and other factors compared with multilateral and bilateral official assistance agencies, they have still played a unique role in fighting against AIDS in China as technical experts, public educators, and civil society supporters. PMID- 16354571 TI - Roles of HIV-1 auxiliary proteins in viral pathogenesis and host-pathogen interactions. AB - Active host-pathogen interactions take place during infection of human immunodeficiency virus type 1 (HIV-1). Outcomes of these interactions determine the efficiency of viral infection and subsequent disease progression. HIV infected cells respond to viral invasion with various defensive strategies such as innate, cellular and humoral immune antiviral mechanisms. On the other hand, the virus has also developed various offensive tactics to suppress these host cellular responses. Among many of the viral offensive strategies, HIV-1 viral auxiliary proteins (Tat, Rev, Nef, Vif, Vpr and Vpu) play important roles in the host-pathogen interaction and thus have significant impacts on the outcome of HIV infection. One of the best examples is the interaction of Vif with a host cytidine deaminase APOBEC3G. Although specific roles of other auxiliary proteins are not as well described as Vif-APOBEC3G interaction, it is the goal of this brief review to summarize some of the preliminary findings with the hope to stimulate further discussion and investigation in this exhilarating area of research. PMID- 16354572 TI - siRNA, miRNA and HIV: promises and challenges. AB - Small interfering RNA (siRNA) and microRNA (miRNA) are small RNAs of 18-25 nucleotides (nt) in length that play important roles in regulating gene expression. They are incorporated into an RNA-induced silencing complex (RISC) and serve as guides for silencing their corresponding target mRNAs based on complementary base-pairing. The promise of gene silencing has led many researchers to consider siRNA as an anti-viral tool. However, in long-term settings, many viruses appear to escape from this therapeutical strategy. An example of this may be seen in the case of human immunodeficiency virus type-1 (HIV-1) which is able to evade RNA silencing by either mutating the siRNA targeted sequence or by encoding for a partial suppressor of RNAi (RNA interference). On the other hand, because miRNA targeting does not require absolute complementarity of base-pairing, mutational escape by viruses from miRNA specified silencing may be more difficult to achieve. In this review, we discuss stratagems used by various viruses to avoid the cells' antiviral si/mi-RNA defenses and notions of how viruses might control and regulate host cell genes by encoding viral miRNAs (vmiRNAs). PMID- 16354573 TI - AIDS and associated malignancies. AB - AIDS associated malignancies (ARL) is a major complication associated with AIDS patients upon immunosuppression. Chronically immunocompromised patients have a markedly increased risk of developing lymphoproliferative disease. In the era of potent antiretrovirals therapy (ARV), the malignant complications due to HIV-1 infection have decreased in developed nations where ARV is administered, but still poses a major problem in developing countries where HIV-1 incidence is high and ARV is still not yet widely available. Even in ARV treated individuals there is a concern that the prolonged survival of many HIV-1 carriers is likely to eventually result in an increased number of malignancies diagnosed. Malignancies that were found to have high incidence in HIV-infected individuals are Kaposi's sarcoma (KS), Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL). The incidence of NHL has increased nearly 200 fold in HIV-positive patients, and accounts for a greater percentage of AIDS defining illness in the US and Europe since the advent of HAART therapy. These AIDS related lymphomas are distinct from their counterparts seen in HIV-1 seronegative patients. For example nearly half of all cases of ARL are associated with the presence of a gamma herpesvirus, Epstein Barr virus (EBV) or human herpesvirus-8 (HHV-8)/ Kaposi's sarcoma associated herpesvirus (KSHV). The pathogenesis of ARLs is complex. B-cell proliferation driven by chronic antigenemia resulting in the induction of polyclonal and ultimately monoclonal lymphoproliferation may occur in the setting of severe immunosuppression. PMID- 16354574 TI - Snapshot of HIV pathogenesis in China. AB - Several reviews have focused on the nature of HIV infection and its spread in various geographical regions of China. In contrast, this review provides a comprehensive update on the prevalence of multiple HIV-1 subtypes, consequent emergence of recombinant and novel forms of HIV-1 in China, and the implications this may have on HIV diversity and the development of effective vaccines. In addition it also examines the dissemination of primary drug resistance in therapy naive patients, as well as co-infections with two other important viruses hepatitis B and C. The main purpose of this review is to provide a current snapshot of HIV-1 pathogenesis in China and possibly shed some light on the future of HIV evolution, and potential challenges for future vaccine and anti retroviral therapeutics against HIV strains in this area. PMID- 16354575 TI - International NeuroAIDS: prospects of HIV-1 associated neurological complications. AB - Neurological complications associated with HIV-1/AIDS are being recognized with a high frequency that parallels the increased number of AIDS cases. The early infiltration by HIV-1 into the nervous system can cause primary and/or secondary neurological complications. The most common neurocognitive disorder is AIDS Dementia Complex (ADC). In developing countries of Asia the three most opportunistic infections are tuberculosis (TB), cryptococcosis, and Pneumocystis carinii pneumonia. Therefore, it is expected that secondary neurological complications due to TB and cryptococcosis will be the most common cause of morbility and mortality in HIV-1/AIDS cases in China. Research of NeuroAIDS in China is necessary to understand the impact and the biology of HIV-1 in the nervous system. Future studies would include, the molecular epidemiology and the description of opportunistic infections associated to HIV-1; the neuropathological description of primary and secondary HIV-1 complications in different groups; the HIV-1 neurotropism and immune response studies for China's unique HIV-1 strains and recombinant forms derived from the nervous system, including experimental models such as the use of transgenic rats; and the study of potential resistant virus, primarily when the anti-retroviral therapy (ART) has not full access in the brain. PMID- 16354576 TI - Errol Crook, MD. PMID- 16354579 TI - Intestinal adaptation in short bowel syndrome. AB - Short bowel syndrome occurs when there is insufficient length of the small intestine to maintain adequate nutrition and/or hydration status without supplemental support. This syndrome most frequently occurs following extensive surgical resection of the intestine, and the extent of adaptation depends on the anatomy of the resected bowel and the amount of bowel remaining. Following resection, the intestinal tissue undergoes morphologic and functional changes to compensate for the lost function of the resected bowel. These changes are mediated by multiple interactive factors, including intraluminal and parenteral nutrients, gastrointestinal secretions, hormones, cytokines, and growth factors, many of which have been well characterized in animal models. The amount of small bowel remaining is the most important predictor of adaptive potential; neither structural nor functional adaptative changes have been demonstrated in humans or animal models with more extreme resections resulting in an end-jejunostomy. The current understanding of these processes has led to the recent use of supplemental hormones, such as growth hormone and glucagon-like peptide 2, in intestinal rehabilitation programs and may lead to the development of pharmacologic agents designed to augment the innate adaptive response. PMID- 16354580 TI - Guanosine triphosphatase activation occurs downstream of calcineurin in cardiac hypertrophy*. AB - There is great interest in deciphering mechanisms of maladaptive remodeling in cardiac hypertrophy in the hope of affording clinical benefit. Potential targets of therapeutic intervention include the cytoplasmic phosphatase calcineurin and small guanosine triphosphate-binding proteins, such as Rac1 and RhoA, all of which have been implicated in maladaptive hypertrophy. However, little is known about the interaction-if any-between these important signaling molecules in hypertrophic heart disease. In this study, we examined the molecular interplay among these molecules, finding that Rho family guanosine triphosphatase signaling occurs either downstream of calcineurin or as a required, parallel pathway. It has been shown that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibition blocks hypertrophy, and we report here that "statin" therapy effectively suppresses small G protein activation and blunts hypertrophic growth in vitro and in vivo. Importantly, despite significant suppression of hypertrophy, clinical and hemodynamic markers remained compensated, suggesting that the hypertrophic growth induced by this pathway is not required to maintain circulatory performance. PMID- 16354581 TI - Current status of integrating information technologies into the clinical research enterprise within US academic health centers: strategic value and opportunities for investment. AB - Little information exists about the incorporation of information technologies (ITs) into clinical research processes within US academic health centers (AHCs). Therefore, we queried a group of 37 leading AHCs regarding their current status and future plans in clinical research IT. The survey specifically inquired about the presence or absence of basic infrastructure and IT support requirements; individual applications needed to support study preparation, study conduct, and its administrative support; and integration of data from basic research, clinical trials, and the clinical information systems increasingly used in health care delivery. Of the 37 AHCs, 78% responded. All strongly agreed that a "state-of-the art" clinical research IT program would be ideal today and will be essential tomorrow. Nonetheless, no AHC currently has an IT solution that even approached this ideal. No AHC reported having all of the essential management foundations (ie, a coherent vision, an overall strategy, a governance structure, and a dedicated budget) necessary to launch and sustain a truly successful implementation of a cohesive clinical research IT platform. Many had achieved breakthroughs in individual aspects of clinical research IT, for example, adverse event reporting systems or consent form templates. However, overall implementation of IT to support clinical research is uneven and insufficient. These data document a substantial gap in clinical research IT investments in leading US AHCs. Linking the clinical research IT enterprise with its clinical operations in a meaningful fashion remains a crucial strategic goal of AHCs. If they are to continue to serve as the "translational research engines" that our society expects, AHCs must recognize this gap and allocate substantial resource deployment to remedying this situation. PMID- 16354582 TI - Osborn (J) wave appearance on the electrocardiogram in relation to potassium transfer and myocardial metabolism during hypothermia. AB - The genesis of the J wave during hypothermia has been attributed to injury current, delayed ventricular depolarization and early repolarization, tissue anoxia, and acidosis. To our knowledge, no studies have addressed the appearance of the J wave in relation to the myocardial K+ transfer and metabolism during hypothermia. Dogs (n = 9) were progressively cooled, blood samples were taken from the aorta and coronary sinus, and myocardial tissue samples were obtained for adenosine triphosphate (ATP), creatine phosphate (CP), and glycolytic intermediate determination. In every instance, the appearance of the J wave was preceded by a net loss of K+ from the myocardium. In one dog, there was no myocardial K+ loss and the J wave was absent. The J wave appeared when the esophageal temperature was between 27 degrees and 24 degrees C (26.6 +/- 0.73 degrees C). At that temperature, the animals were hypotensive and bradycardic, but arterial oxygen partial pressure, carbon dioxide partial pressure, and pH were within the physiologic range at that temperature. The myocardial ATP and CP from the hypothermic dogs was lower compared with the value obtained from dogs at 37 degrees C (p < .025 and p < .005, respectively). The levels of the glycolytic intermediates, fructose-1,6-diphosphate, dihydroxyacetone phosphate, and pyruvate, were lower and the level of lactate was higher compared with those from the normothermic dogs (not significant; p < .007, p < .02, p < .001, respectively). These findings suggest that the appearance of the J wave on electrocardiography during cooling is a result of depression of the metabolic process concerned with maintenance of the partition of ions across the cell membrane, as evidenced by decreased myocardial energy content and K+ loss during the hypothermic state. PMID- 16354586 TI - Activating mutations and/or expression levels of tyrosine kinase receptors GRB7, RAS, and BRAF in testicular germ cell tumors. AB - Amplification and/or overexpression of genes encoding tyrosine kinase receptors KIT and ERBB2 have been reported in testicular germ cell tumors (TGCTs). These receptors can bind the adaptor molecule GRB7 encoded by a gene adjacent to ERBB2 at 17q12, a region also frequently gained in TGCTs. GRB7 binding may be involved in the activation of RAS signaling and KRAS2 maps to 12p, which is constitutively gained in TGCT and lies within a minimum overlapping region of amplification at 12p11.2-12.1, a region we have previously defined. RAS proteins activate BRAF, and activating mutations of genes encoding these proteins have been described in various tumors. Here we determine the relationships between expression levels and activating mutations of these genes in a series of 65 primary TGCTs and 4 TCGT cell lines. High levels of expression and activating mutations in RAS were mutually exclusive events, and activating mutations in RAS were only identified in the seminoma subtype. Mutations in BRAF were not identified. Increased ERBB2 expression was associated with differentiated nonseminoma histology excised from lymph nodes postchemotherapy. Mutation, elevated expression, and correlations between expression levels of KRAS2, GRB7, and KIT are consistent with their involvement in the development of TGCTs. PMID- 16354587 TI - Efficacy of deguelin and silibinin on benzo(a)pyrene-induced lung tumorigenesis in A/J mice. AB - We evaluated deguelin and silibinin in A/J mice treated with the tobacco-specific carcinogen benzo(a)pyrene (BP) for their ability to inhibit pulmonary adenoma formation and growth. Animals were treated with either deguelin (5.0 or 10.0 mg/kg body weight, by gavage) or silibinin at doses of 0.05% and 0.1% in the diet, approximately 10 days before a single intraperitoneal dose of BP. We found that oral administration of deguelin reduced tumor multiplicity by 56% and tumor load by 78%, whereas silibinin treatment at doses of 0.05% and 0.1% in the diet did not show any significant efficacy on either tumor multiplicity or tumor load. The result indicates that deguelin significantly inhibits pulmonary adenoma formation and growth in A/J mice. Finding new and effective agents that can prevent lung cancer is urgently needed because cancer of the lungs remains the principal cause of cancer deaths in the United States and because effective chemoprevention of this cancer type remains elusive. Thus, deguelin appears to be a promising new preventive agent for lung cancer and may be considered for further studies in other animal models and in clinical trials. PMID- 16354585 TI - A review of the past, present, and future directions of neoplasia. PMID- 16354588 TI - Expression of axl in lung adenocarcinoma and correlation with tumor progression. AB - We used the Transwell system to select highly invasive cell lines from minimally invasive parent cells, and we compared gene expression in paired cell lines with high and low invasive potentials. Axl was relatively overexpressed in the highly invasive cell lines when compared with their minimally invasive counterparts. However, there is only limited information about the role of Axl in cancer invasion. The biologic function of Axl in tumor invasion was investigated by overexpression of full-length Axl in minimally invasive cells and by siRNA knockdown of Axl expression in highly invasive cells. Overexpression of Axl in minimally invasive cells increased their invasiveness. siRNA reduced cell invasiveness as Axl was downregulated in highly invasive cells. We further investigated the protein expression of Axl by immunohistochemistry and its correlation with clinicopathologic features. Data from a study of 58 patient specimens showed that Axl immunoreactivity was statistically significant with respect to lymph node status (P < .0001) and the patient's clinical stage (P < .0001). Our results demonstrate that Axl protein kinase seems to play an important role in the invasion and progression of lung cancer. PMID- 16354589 TI - Construction of a novel constitutively active chimeric EGFR to identify new targets for therapy. AB - Tumor cells and tumor-associated endothelial cells express activated epidermal growth factor receptor (EGFR) due to production of EGF-related ligands in the tumor microenvironment. To investigate the effect of perpetual EGFR activation on endothelial cells, we developed a novel method to generate constitutively active EGFR. We fused the entire intracellular domain of the EGFR to the N-terminus of the CD3zeta component of the T-cell receptor signaling complex. Expression of the chimeric receptor CD3-EGFR in EGFR-deficient human embryonic kidney cells resulted in ligand-independent sustained EGFR phosphorylation and in the induction of Akt, mitogen-activated protein kinase, and signal transducer and activator of transcription 3 (Stat3). Next, CD3-EGFR was stably expressed in murine brain endothelial cells where it signaled for the initiation of angiogenic programs, Stat3 activation, and continuous proliferation. A comparison between brain endothelial cells encoding CD3zeta and CD3-EGFR revealed that proangiogenic phenotype was modulated by the intracellular effector Stat3 and that suppression of this downstream target with the EGFR tyrosine kinase inhibitor PKI166 could revert this phenotype. Thus, our results validate the use of chimeric constitutively active receptors to replicate critical features observed in pathophysiological processes that can expedite the identification of novel therapeutic agents targeting EGFR activation and function. PMID- 16354590 TI - BJ-TSA-9, a novel human tumor-specific gene, has potential as a biomarker of lung cancer. AB - Using bioinformatics, we have identified a novel tumor-specific gene BJ-TSA-9, which has been validated by Northern blot analysis and reverse transcription polymerase chain reaction (RT-PCR). BJ-TSA-9 mRNA was expressed in 52.5% (21 of 40) of human lung cancer tissues and was especially higher in lung adenocarcinoma (68.8%). To explore the potential application of BJ-TSA-9 for the detection of circulating cancer cells in lung cancer patients, nested RT-PCR was performed. The overall positive detection rate was 34.3% (24 of 70) in peripheral blood mononuclear cells (PBMCs) of patients with various types of lung cancers and was 53.6% (15 of 28) in PBMCs of lung adenocarcinoma patients. In combination with the detection of two known marker genes SCC and LUNX, the detection rate was increased to 81.4%. A follow-up study was performed in 37 patients after surgical removal of tumor mass. Among nine patients with persistent detection of two to three tumor marker transcripts in PBMCs, six patients had recurrence/metastasis. In contrast, 28 patients with transient detection of one tumor marker or without detection of any tumor marker were all in remission. Thus, BJ-TSA-9 may serve as a marker for lung cancer diagnosis and as a marker, in combination with two other tumor markers, for the prediction of the recurrence and prognosis of lung cancer patients. PMID- 16354591 TI - Regulation of the pathological vasculature of malignant astrocytomas by angiopoietin-1. AB - Malignant astrocytomas are the most common and highly vascularized of all primary adult brain tumors. The histopathological hallmarks of malignant astrocytomas are microvascular proliferation and formation of vascular entities, which are referred to as "glomeruloid bodies." The significance of glomeruloid bodies and the molecular mechanisms driving the abnormal vascular architecture in malignant astrocytomas are not understood. We have observed that overexpression of angiopoietin-1 (Ang1) in both subcutaneous and intracranial xenograft models of malignant astrocytomas reproduces many of the vascular features of these tumors, including glomeruloid bodies. To confirm that the formation of glomeruloid bodies was directly dependent on Ang1, we performed experiments where levels of Ang1 expression were regulated under tetracycline control, and we found a direct correlation between levels of Ang1 expression and the occurrence of glomeruloid bodies in xenografts. Additionally, we inhibited the action of Ang1 by blocking its cognate receptor Tie2, and we found that the formation of glomeruloid bodies was inhibited. Collectively, these results support our hypothesis that Ang1 is a key molecular regulator of pathological vascularization characteristic of malignant astrocytomas. PMID- 16354592 TI - Uroplakin Ib gene transcription in urothelial tumor cells is regulated by CpG methylation. AB - Uroplakin Ib is a structural protein on the surface of urothelial cells. Levels of uroplakin Ib mRNA are dramatically reduced or absent in many transitional cell carcinomas, but the molecular mechanisms responsible remain undetermined. Previously, we showed that loss of uroplakin Ib expression correlated with CpG methylation of Sp1/NFkappaB-binding motifs within the proximal promoter. In this study, we show that reporter activity was completely blocked by the methylation of three CpG pairs in this promoter region. Gel shift analysis using purified proteins or nuclear extracts showed that Sp1 and NFkappaB bound to motifs encompassing two of the three CpG pairs. Interestingly, the methylation of these two CpG sites did not prevent the binding of proteins to the promoter in gel shift analyses. Additionally, mutation of these two CpGs did not affect reporter activity, but mutation of 6-bp fragment spanning each CpG partially inhibited reporter activity, suggesting that these sites were functional. A requirement for both Sp1 and NFkappaB in regulating reporter activity was confirmed in transfection experiments using plasmids expressing individual proteins. Our data suggest that the methylation of specific CpG sites can silence the uroplakin Ib promoter, at least in part, by blocking the binding of Sp1 and NFkappaB, although other factors may be involved. PMID- 16354593 TI - Combination of proteasomal inhibitors lactacystin and MG132 induced synergistic apoptosis in prostate cancer cells. AB - The proteasome inhibitor Velcade (bortezomib/PS-341) has been shown to block the targeted proteolytic degradation of short-lived proteins that are involved in cell maintenance, growth, division, and death, advocating the use of proteasomal inhibitors as therapeutic agents. Although many studies focused on the use of one proteasomal inhibitor for therapy, we hypothesized that the combination of proteasome inhibitors Lactacystin (AG Scientific, Inc., San Diego CA) and MG132 (Biomol International, Plymouth Meeting, PA) may be more effective in inducing apoptosis. Additionally, this regimen would enable the use of sublethal doses of individual drugs, thus reducing adverse effects. Results indicate a significant increase in apoptosis when LNCaP prostate cancer cells were treated with increasing levels of Lactacystin, MG132, or a combination of sublethal doses of these two inhibitors. Furthermore, induction in apoptosis coincided with a significant loss of IKKalpha, IKKbeta, and IKKgamma proteins and NFkappaB activity. In addition to describing effective therapeutic agents, we provide a model system to facilitate the investigation of the mechanism of action of these drugs and their effects on the IKK-NFkappaB axis. PMID- 16354594 TI - A population of HLA-DR+ immature cells accumulates in the blood dendritic cell compartment of patients with different types of cancer. AB - Dendritic cell (DC) defects are an important component of immunosuppression in cancer. Here, we assessed whether cancer could affect circulating DC populations and its correlation with tumor progression. The blood DC compartment was evaluated in 136 patients with breast cancer, prostate cancer, and malignant glioma. Phenotypic, quantitative, and functional analyses were performed at various stages of disease. Patients had significantly fewer circulating myeloid (CD11c+) and plasmacytoid (CD123+) DC, and a concurrent accumulation of CD11c( )CD123(-) immature cells that expressed high levels of HLA-DR+ immature cells (DR(+)IC). Although DR(+)IC exhibited a limited expression of markers ascribed to mature hematopoietic lineages, expression of HLA-DR, CD40, and CD86 suggested a role as antigen-presenting cells. Nevertheless, DR(+)IC had reduced capacity to capture antigens and elicited poor proliferation and interferon-gamma secretion by T-lymphocytes. Importantly, increased numbers of DR(+)IC correlated with disease status. Patients with metastatic breast cancer showed a larger number of DR(+)IC in the circulation than patients with local/nodal disease. Similarly, in patients with fully resected glioma, the proportion of DR(+)IC in the blood increased when evaluation indicated tumor recurrence. Reduction of blood DC correlating with accumulation of a population of immature cells with poor immunologic function may be associated with increased immunodeficiency observed in cancer. PMID- 16354599 TI - Hyperoxic ventilation enables hemodilution beyond the critical myocardial hemoglobin concentration. AB - BACKGROUND: When initiated in anemic hypoxia, hyperoxic ventilation (ventilation with pure O2, FiO2 1.0, HV) reverses hypoxia-induced ECG-changes and enables survival for several hours. The quantification of the HV-induced gain in anemia tolerance and particularly the Hb-equivalent of HV in this situation are unknown. METHODS: Nine anaesthetized pigs were hemodiluted under normoxia (FiO2 0.21) by exchange of whole blood for hydroxyethyl starch (HES) until predefined, ischemia associated ECG-changes occurred (timepoint Hb(crit)). From that time on all animals were ventilated with 100% O2 (FiO2 1.0). In the case of disappearance of the ECG changes with onset of HV, the animals were further hemodiluted until ECG changes reoccurred. RESULTS: HV initiated in anemic hypoxia (Hb 2.3 +/- 0.2 g/dl) improved ECG-readings of all animals, and allowed for a further exchange of 14 +/ 11 ml/kg blood until ECG-changes reoccurred at Hb 1.2 +/- 0.4 g/dl. CONCLUSION: HV initiated in anemic hypoxia creates a margin of safety for myocardial tissue oxygenation and thus further increases anemia tolerance. The Hb equivalent of HV in this situation amounts to approximately 1g/dl. PMID- 16354595 TI - HLA-DR+ immature cells exhibit reduced antigen-presenting cell function but respond to CD40 stimulation. AB - Dendritic cells (DC) have been implicated in the defective function of the immune system during cancer progression. We have demonstrated that patients with cancer have fewer myeloid (CD11c+) and plasmacytoid (CD123(hi)) DC and a concurrent accumulation of CD11c(-)CD123- immature cells expressing HLA-DR (DR(+)IC). Notably, DR(+)IC from cancer patients have a reduced capacity to stimulate allogeneic T-cells. DR(+)IC are also present in healthy donors, albeit in smaller numbers. In this study, we assessed whether DR(+)IC could have an impact on the immune response by comparing their function with DC counterparts. For this purpose, DR(+)IC and DC were purified and tested in the presentation of antigens through major histocompatibility complex (MHC) II and MHC-I molecules. DR(+)IC were less efficient than DC at presenting antigens to T-cells. DR(+)IC induced a limited activation of T-cells, eliciting poor T-helper (Th) 1 and preferentially inducing Th2-biased responses. Importantly, despite DR(+)IC's poor responsiveness to inflammatory factors, in samples from healthy volunteers and breast cancer patients, CD40 ligation induced phenotypic maturation and interleukin 12 secretion, in turn generating more efficient T-cell responses. These data underscore the importance of inefficient antigen presentation as a mechanism for tumor evasion and suggest an approach to improve the efficacy of DC-based immunotherapy for cancer. PMID- 16354598 TI - Effects of soluble TNF receptor II (sTNF-RII), IL-1 receptor antagonist (IL-1ra), tumor load and hypermetabolism on malnutrition in children with acute leukemia. AB - OBJECTIVE: Soluble tumor necrosis factor receptor II (sTNF-RII) and interleukin-1 receptor antagonist (IL-1ra) might modulate nutritional status in acute leukemia since they are inhibitors of tumor necrosis factor-alpha and interleukin-1 that can induce tissue wasting. On the other hand, tumor load and hypermetabolism may induce malnutrition. We determined whether serum levels of sTNF-RII and IL-1ra are upregulated to prevent overt malnutrition and whether tumor load and hypermetabolism induce overt malnutrition. METHODS: We examined 31 children with newly diagnosed acute leukemia and correlated sTNF-RII, IL-1ra, tumor load and energy expenditure to anthropometric characteristics (weight, weight for height, height, body mass index, fat free mass) and serum protein concentrations (albumin, transferrin, prealbumin). As controls, 68 healthy children were examined for anthropometric characteristics; 33 healthy controls were included for cytokine analysis and biochemical indices. RESULTS: We found no correlations between sTNF-RII, IL-1ra, tumor load and energy expenditure and anthropometric characteristics or protein concentrations. Mean sTNF-RII level was significantly, mean IL-1ra level slightly increased (223% and 113% of the controls). 29% of the children had a high tumor load (> 100.000/microl white blood cells) and 53% had hypermetabolism (resting energy expenditure > 110% of predicted). Anthropometric characteristics were similar to those in controls, however, serum protein concentrations were decreased. CONCLUSION: sTNF-RII and IL-1ra are upregulated in children with leukemia and may therefore prevent overt malnutrition. Tumor load and hypermetabolism do not induce overt malnutrition. The children presented with an early stage of malnutrition as evidenced by low serum protein concentrations but normal anthropometric characteristics. PMID- 16354600 TI - Monitoring of expiratory flow rates and lung volumes during a high altitude expedition. AB - BACKGROUND: Data on lung volumes and changes in flow-volume spirometry at high altitude are few and do not provide comprehensive assessment of the occurring changes. This study characterizes alterations of the forced expiratory flow volume curve (FEFV-curve) and lung volumes at increasing altitude. METHODS: FEFV curve and lung volumes at increasing altitude were characterized by daily assessment of peak expiratory flow (PEF), forced expiratory volume in one second (FEV1), forced vital capacity (FVC) and maximal expiratory flow rates (MEF 25, 50, 75) at 25%, 50% and 75% of the FEFV-curve with a portable spirometer (turbinometric method) three times a day during an expedition to Mustagh Ata (7545m) in 15 healthy mountaineers. RESULTS: With increasing altitude FVC and FEV1 were reduced by up to 25% (74.8% / 74.6% of baseline) and MEF25 was reduced to 81.5% of baseline values. PEF initially increased up to 4451m and returned to baseline values above 5000m. After descent below 2000m, all values normalized within one day. There were weak negative correlations between AMSS and FEV1, FVC and PEF (r = -0.23, p<0.001). CONCLUSIONS: We found increasing pulmonary restriction at high altitude without a marked reduction of PEF. Assessment of the FEFV-curve at high altitudes with a portable spirometer is a practical method reflecting the true field situation and may provide clinically relevant information (impending pulmonary edema). PMID- 16354601 TI - Immunohistochemical examination of cytological differentiation in osteosarcomas. AB - In this immunohistochemical examination, the expression of Runx2, Notch1, Delta and Osteopontin peptides were detected in neoplastic cells in 10 Japanese cases of osteosarcoma. Immunohistochemically, Runx2 peptide expression appeared in the cytoplasm of almost all neoplastic cells of the 10 cases examined. However, Notch1 peptide expression appeared in the cytoplasm of neoplastic cells in the localized and comparatively well-differentiated area of osteosarcoma, which osteoblastic and chondroblastic containing osteoid and/or chondroid tissues. No expression of Notch1 peptide was detected in the fibroblastic and poorly differentiated areas. Delta peptide appearance was nearly the same pattern of Notch1 peptide. Expression of Osteopontin peptide appeared in almost all cells and the strength expression was shown in the area of comparatively well differentiated tissues. Therefore, these results suggest that Runx2, Notch1, and Delta peptides are closely related to cytological differentiation or acquisition of tissue specific characteristics in neoplastic cells in osteosarcomas. PMID- 16354602 TI - Bone mineral density changes and bone turnover in thyroid carcinoma patients treated with supraphysiologic doses of thyroxine. AB - The aim of this one-year prospective study was to determine whether longterm thyroxine treatment is a risk factor for elevated bone turnover, loss of bone mass and subsequent development of osteoporosis. Premenopausal women (N = 19), and men (N = 9) suffering from differentiated thyroid gland carcinoma in the mean age of 39.0 +/- 8.0 years and 41.8 +/- 10.0 years were investigated. All of them had undergone a total thyroidectomy and subsequent thyroxine therapy. The duration of the TSH-suppressive therapy prior to the the beginning of our study was 9.4 +/- 6.4 years in the female and 8.1 +/- 6.0 years in the male group. The prospective observation was performed by dual X-ray absorptiometry (DXA) at the spine and the femoral neck and by single-photon absorptiometry (SPA) at the distal radius. Laboratory testings included thyroid hormones T3, T4 and TSH, serum calcium, phosphate and PTH, and urinary calcium and phosphate from spontaneous and 24-hour urine samples. Markers of bone formation (osteocalcin, alkaline phosphatase and PICP) and resorption (Ca/Cr and ICTP) were determined. Statistically significant loss of bone mass was observed only on the distal radius in males (p<0.05). At the lumbar spine and femoral neck, only a minor bone loss was registered in a small number of patients. Almost 50 % of the females showed values above the reference range. In more than 30 % of the females, and smaller number of male patients, ICTP values ranged above the reference range, corresponding to elevated bone turnover. These two variables exhibited a slight correlation with bone density at the measured skeletal areas, mostly considering the male group. The results are a proof that accelerated bone turnover and subsequent bone loss occurs during TSH-suppressive thyroxine therapy. In future prospective studies a prolonged time of observation will be necessary, as well as to increase the number of studied patients, in order to better assess the relative risk of osteoporosis in patients undergoing TSH-suppressive treatment more precisely. PMID- 16354603 TI - GNB3 C825T polymorphism and response to anti-retroviral combination therapy in HIV-1-infected patients--a pilot study. AB - METHODS: Analyses were based on a cohort of 55 HIV infected patients whose treatment was changed due to therapy failure. Viral load, CD4+ and CD8+ cell counts were determined before therapy and after 8 weeks and 16-32 weeks, respectively. GNB3 genotyping was performed using Pyrosequencing. Chemotaxis of purified CD4+ cells was quantified in a Boyden chamber using stromal cell-derived factor 1alpha (SDF-1alpha) as a stimulus. RESULTS: Age gender, route of infection, treatment, and baseline values for viral load and CD4+ and CD8+ cell counts were not associated with GNB3 genotypes (10 TT, 24 TC, 21CC). After 8 weeks of treatment viral load was lowest in TT genotypes (log copies/ml: TT: 1.88 +/- 0.32; TC: 2.49 +/- 0.24; CC: 3.06 +/- 0.22; p = 0.039) and this trend tended to persist until final visit. Unexpectedly, TT genotypes had lowest CD4+ cell counts at final visit (TT: 165.3 +/- 34.0; TC: 414.4 +/- 72.1; CC: 441.5 +/-70.9; p = 0.047). SDF-1alpha-stimulated chemotaxis was reduced in CD4+ cells from HIV-1 infected patients compared to healthy controls. Still cells from TT genotypes displayed strongest chemotaxis. CONCLUSIONS: This pilot study suggests that the GNB3 C825T polymorphism is associated with short-term success of HAART treatment in HIV-1-infected patients. PMID- 16354604 TI - Traumatic pneumobilia after cardiopulmonary resuscitation. AB - Pneumobilia is a rare pathological finding, which denotes an abnormal connection between the gastrointestinal and the biliary tract. In the absence of surgically created anastomosis between the bowel and the bile duct, the most common causes for pneumobilia are gallstone obstruction, endoscopic interventions, or emphysematous cholecystitis. We present this case of a middle-aged multiple injured male who developed traumatic pneumobilia after cardiopulmonary resuscitation. We suppose that chest compression in combination with a sphincter of Oddi (SO) dysfunction forced intraluminal air retrograde through the SO into the biliary tract, since intraabdominal injury as well as former biliary pathology, inflammation, or biliary-enteric fistula were excluded. PMID- 16354605 TI - Symptoms and signs of an acute myocardial ischemia caused by chemotherapy with Paclitaxel (Taxol) in a patient with metastatic ovarian carcinoma. AB - INTRODUCTION: Paclitaxel (Taxol) is an anticancer agent used for the treatment of breast and ovarian cancer. The major side effects are bone marrow suppression, alopecia, polyneuropathy and cardiac toxicity like bradycardia, myocardial infarction, congestive heart failure and cardiac death. SETTING: Intensive care unit (ICU) of a university hospital. PATIENT: We report on a 58-years-old woman with a metastatic ovarian carcinoma who had chest pain, nausea and collapse during their first Taxol infusion. The infusion was stopped and the patient was submitted to the intensive care unit (ICU) to exclude an acute coronary syndrome. RESULTS: The electrocardiography (ECG) showed a third-degree heart block and ST elevation in II, III and avF. In the initial and in the control laboratory investigation values of cardiac enzymes (creatininkinase and Troponine T) remained normal. The control ECG after 30 minutes turned back to normal. After one day the patient was submitted back to a normal ward. CONCLUSION: Symptomatic bradyarrhythmia and clinical sign of an myocardial infarction are rare but important cardiac side effects in patients treated with Taxol. Those patients should be under intensive care unit until patients conditions improve and acute myocardial ischemia has been excluded. PMID- 16354606 TI - Expression of Kir2.1 channel during differentiation of human macrophages into foam cells. AB - OBJECTIVE: Detected in non-transformed bone marrow-derived macrophages (BMDM) and identified as one of the key channels in modulating macrophage proliferation, activation and apoptosis, Kir2.1 channel is also characterized to play a crucial role in cell differentiation. The purpose of this study was to investigate the expression of Kir2.1 channel mRNA and protein during human monocyte-derived macrophage differentiation into foam cells. METHODS: Human peripheral blood monocytes were isolated from healthy male volunteers by density gradient centrifugation and then by adherence method. The macrophages identified as a homogeneous population of adherent cells were obtained after 5 days of culture. Expression of Kir2.1 channel during human macrophage differentiation into foam cells was investigated by RT-PCR, Western blotting and immunocytochemistry, respectively. RESULTS: After incubation of the macrophages with 30 mg/L OxLDL at 37 degrees C for 60 h, the cells were obviously enlarged in size and numerous red lipid granules observed under optical microscope. The cellular contents of the total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) were markedly increased from 54.79+/-28.304 mg/g, 47.968+/-26.787 mg/g and 6.822+/ 3.437 mg/g to 229.775+/-57.453 mg/g, 96.241+/-24.003 mg/g and 133.535+/-36.292 mg/g, respectively; the CE/TC ratio rose from (14.437+/-6.781)% to (57.946+/ 3.507)% (n=7, P<0.05), suggesting the phenotype of foam cells. However, there was no significant difference in the relative expression of Kir2.1 channel mRNA between the macrophages and foam cells [(59.074+/-10.566)% vs (46.98+/-12.527)%, n=5, P>0.05], nor was there significant difference in the relative expression of Kir2.1 channel protein between them [(60.527+/-18.621)% vs (50.243+/-11.583)%, n=6, P>0.05]. CONCLUSION: Incubation of human monocyte-derived macrophages with 30 mg/L OxLDL for 60 h induces the differentiation of the cells into foam cells, but the expression of Kir2.1 channel does not change obviously. PMID- 16354607 TI - Selection of femoral prosthesis in total hip replacement for ankylosing spondylitis. AB - OBJECTIVE: To investigate the implication of morphological changes of the proximal femur in femoral prosthesis selection in total hip replacement for treatment of ankylosing spondylitis (AS). METHODS: Twenty-four patients (36 hips) admitted for cementless total hip replacement (THR) because of AS were enrolled in this study along with the control group of 30 patients (45 hips) without AS. Measurement of the Singh index, canal flare index, morphological index of the cortex, cortical index and stem-canal fit was performed in the two groups on the basis of roentgenograms. RESULTS: The AS patients had more serious osteoporosis in their proximal femur. There were significant differences between the two groups in Singh index, canal flare index, morphological index of the cortex and stem-canal fit (P<0.05), but not in the cortical index (P>0.05). The AS patients were found to have severe osteoporosis in the upper femur and massive spongy bone loss, so that the morphology of the canal was almost orthostatic in the upper femur because of the thinned femoral cortex. CONCLUSION: Cemented prosthesis is recommended for AS patients with severe osteoporosis, who should be treated with revised THR to ensure good fit of the prosthesis to the canal. Cementless prosthesis, however, can still achieve good therapeutic effect in AS patients without significant morphologic changes in the proximal femurs. Computer-aided design and manufacture of individualized prosthesis better suits the AS patients than the currently available prosthesis. PMID- 16354608 TI - [Liposome-mediated human CD40 gene transfection and human umbilical vein endothelial ECV-304 cells]. AB - OBJECTIVE: To construct an eukaryotic expression vector containing human CD40 gene for its efficient, continuous and stable expression in human umbilical vein endothelial ECV-304 cells. METHODS: The recombinant plasmid pUCD40 was digested with endonucleases to obtain human CD40 gene fragment, which was cloned into pCDNA3.1 vector to construct recombinant eukaryotic expression vector pCDNA3.1(+)/CD40. The recombinant vector was identified by enzyme digestion before introduced into ECV-304 cells via liposome, with the positive cell clones selected with G418. The stable transfection and expression of CD40 in ECV-304 cells were identified by reverse transcription (RT)-PCR, Western blotting and flow cytometry, respectively. RESULTS: Enzyme digestion analysis showed that target gene had been cloned into the recombinant vector. The transfected ECV-304 cells successfully expressed human CD40 as determined by RT-PCR and Western blotting, and 95% of the cells were CD40-positive as shown by flow cytometry. CONCLUSION: The recombinant eukaryotic expression vector pCDNA3.1(+)/CD40 has been successfully constructed, which is capable of stable transfection and expression of CD40 in ECV-304 cells to facilitate further investigation of the roles of CD40 molecule in antiatherosclerotic drug development. PMID- 16354609 TI - [Expression of DNA transcription- and repair-related genes in cisplatin-resistant human ovarian carcinoma cell line COC1/DDP]. AB - OBJECTIVE: To study the molecular mechanism underlying cisplatin resistance in ovarian carcinoma by detecting the expressions of DNA transcription- and repair related genes in cisplatin-resistant human ovarian carcinoma COC1 cell line. METHODS: The differential expression of DNA transcription- and repair-related genes between the parental COC1 and cisplatin-resistant COC1/DDP cell line was determined using cDNA microarray. RESULTS AND CONCLUSION: Compared with COC1 cells, 143 genes in COC1/DDP cells showed significant differential expression, among which 20 were DNA transcription- and repair-related genes including 13 significantly up-regulated genes and 7 down-regulated ones. Abnormality of DNA transcription and repair might be involved in the development of cisplatin resistance in COC1/DDP cells. PMID- 16354610 TI - [Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells]. AB - OBJECTIVE: To construct the eukaryotic expression vector for HPC2 for expression in HEK293 cells. METHODS: HPC2 from pcDNA3/HPC2 were inserted into the flag tagged vector pcDNA3-flag by subcloning method. The recombinant plasmid pcDNA3 flag/HPC2 was then transfected into HEK293 cells using a routine lipofectamine method. The cell lysate was used for Western blotting to examine the expression of the target protein. RESULTS AND CONCLUSION: Double restriction enzyme digestion and DNA sequencing indicated successful construction of the eukaryotic expression vector for HPC2 and the fusion protein was highly expressed in HEK293 cells, which provides an important basis for functional study of HPC2. PMID- 16354611 TI - [Effect of magnetic mitomycin C nanoparticles on proliferation and apoptosis of L 02 cells in vitro]. AB - OBJECTIVE: To investigate the effects of mitomycin magnetic polybutylcyanoacrylate nanoparticles (MMC-MPBCA- NP), magnetic polybutylcyanoacrylate nanoparticles (MPBCA-NP) and magnetic fluids (MF) and MMC on the proliferation and apoptosis of normal human liver cell strain (L-02 cells) in vitro. METHODS: The effect of MMC, MMC-MPBCA-NP, MPBCA-NP, and MF on the apoptosis and cell cycle of L-02 cells was studied by flow cytometry. The changes in the cell proliferation in response to treatments of the preparations and agents were examined by MTT colorimetric assay. RESULTS: MMC, MMC-MPBCA-NP, MPBCA NP produced inhibition of the cell proliferation to different degrees but MF failed to result in cell proliferation inhibition. The results of MTT assay showed that MM, MPBCA-NP and MMC-MPBCA-NP all possessed concentration-dependent cytotoxicity, which increased with the concentration increment in comparison with the blank control. CONCLUSION: The descending order of the 4 agents in their effect of L-02 cell strain growth inhibition is MF >MPBCA-NP>MMC-MPBCA-NP>MMC. MMC and MMC-MPBCA-NP can inhibit the proliferation and cause cell cycle changes of L-02 cells, while MPBCA-NP inhibits the cell proliferation without inducing cell cycle alterations. PMID- 16354612 TI - [Expression of HLA-G protein in trophoblast cells]. AB - OBJECTIVE: To investigate the expression of human leucocyte antigen protein G (HLA-G) in different trophoblast cells and different stages of pregnancy. METHODS: The expression of HLA-G protein in normal placenta and trophoblasts of different trimesters was detected using immunohistochemical method (SP). RESULTS: HLA-G protein expression exhibited spatio-temporal changes, which located in the extravillous trophoblast (EVT) and was higher in the placenta of the first and second trimesters while lower in the third trimester (P<0.01). CONCLUSION: HLA-G protein expression in different stages of pregnancy and different trophoblasts may be related to the controlled invasion of the trophoblast. PMID- 16354613 TI - [Effect of chronic enhanced external counterpulsation on arterial endothelial cells of porcine with hypercholesteremia]. AB - OBJECTIVE: To verify whether chronic enhanced external counterpulsation (EECP) may repair artery endothelial cells (ECs) damage resulted from hypercholesteremia in pigs. METHODS: EECP was performed for 36 hours in pigs with hypercholesteremia and the left descending artery (LDA) was isolated for scanning electron microscopy (SEM). The ECs were collected from the thoracic aorta and analyzed by proteomic technique. RESULTS: The ECs of hypercholesteremia pigs showed irregular arrangement with obvious desquamation of coronary ECs and formation of atherosclerotic plaque, and these lesions were milder in EECP-treated pigs. Six over-expressed proteins were detected in the endothelial cells in EECP-treated pigs in comparison with those of the hypercholesteremia pigs. CONCLUSION: Chronic EECP helps restore cell morphology and repair functional damage of ECs resulted from hypercholesteremia by regulating endothelial protein expressions, and consequently improves cell adhesion and intracellular metabolism and reduces EC apoptosis. PMID- 16354614 TI - [A new method for static intensity-modulated radiation therapy]. AB - OBJECTIVE: To develop a new method for static intensity-modulated radiation therapy (IMRT) possible for implementation in smaller hospitals by incorporating accurate location equipments and treatment planning system in the available equipments for general radiotherapy. METHODS: Based on the techniques of conformal radiotherapy, a new split method for fabricating the three-dimensional physical compensator was developed to achieve IMRT. RESULTS: Experiment with the new method showed that the fabricated compensator could achieve good match between the high dosage shape in the target area and the dimensions of the tumor, and allowed adjustment of the dosage distribution according to the therapeutic requirement. CONCLUSION: This simple and feasible method allows cost-effect application of IMRT in smaller hospitals. PMID- 16354615 TI - [Rat model of Klebsiella pneumoniae-induced severe pneumonia]. AB - OBJECTIVE: To explore the diagnostic criteria and physiopathological features of severe bacterial pneumonia in rats. METHODS: A total of 60 Sprague Dawleg rats were randomly divided into 3 groups, namely the model group (n=24), observation group (n=24), and control group (n=12). The rats in the former two groups were intratracheally instilled with Klebsiella pneumoniae suspension at different doses, while those in the control group received intratracheal administration of 1 ml saline. On the 2nd, 4th, and 6th days after intratracheal instillation, 1/3 of the rats in each group were killed to determine the hemodynamics, arterial blood gas (ABG), peripheral hemogram, leukocytes and neutrophils in the bronchoalveolar lavage fluid (BALF), followed by pathological examination of the lung. RESULTS: Obvious hemodynamic changes occurred in the rats of the model group on the 5th day and ABG changes appeared on the 4th day. The changes in peripheral hemogram, leukocytes and neutrophils in the BALF, and lung pathology of the rats in the model group were more obvious than those in the observation group on each time point. As the time prolonged, the changes in the hemodynamics, ABG, peripheral hemogram, leukocytes and neutrophils in the BALF and pathology of the lung exacerbated in the model group and observation group, but all these indices in the control group remain unchanged after bacterial inoculation. CONCLUSION: The severity of pneumonia in rats increases with the dose increment of the administered Klebsiella pneumoniae suspension, which mimics the pathological changes in severe bacterial pneumonia in human. PMID- 16354616 TI - The effect of L-thyroxine supplementation on retinal vascular development in neonatal rats. AB - PURPOSE: Thyroxine (T4) plays a role in neuroretinal maturation, but little is known regarding its role in retinal vascularization. The neonatal rat retina is incompletely vascularized at birth, providing a model for the human premature infant retina and for retinopathy of prematurity (ROP). We hypothesized that T4 supplementation would accelerate vascular development of normal neonatal rat retina. METHODS: Two hundred twenty Sprague-Dawley rats were raised in litters of 10 in room air and received either 0.05 microg/g, 0.5 microg/g, or 1.0 microg/g of intraperitoneal T4 or saline control beginning on day 1 of life for 3 days, 7 days, or 3 days followed by 4 days recovery. Rats were sacrificed on either day 4 or day 8 of life. Left eyes were fixed, retinae dissected and ADPase-stained. Flat mounted retinae were digitized and total retinal areas and retinal vascular density were evaluated in a masked manner. Serum T4, thyroid stimulating hormone (TSH), and insulin-like growth factor-1 (IGF-1) were measured at each time point. RESULTS: Retinal vascular density was reduced in animals receiving daily 1 microg/g T4 compared with saline controls after 3 days of T4 (16.8 +/- 1.4 vessels/mm vs. 18.3 +/- 1.3 vessels/mm, p = 0.04) and 7 days of T4 (14.4 +/- 1.3 vessels/mm vs. 16.8 +/- 1.1 vessels/mm, p < 0.0006). However, retinal vascular density returned to normal after 3 days of treatment and 4 days of recovery. Vascularized retinal area was reduced in animals receiving 1 microg/g T4 for 3 days followed by 4 days recovery compared with saline controls (85 +/- 6% vs. 92 +/- 3%, p = 0.002). At lower doses of T4 (0.05 microg/g and 0.5 microg/g for 3 or 7 days) and at 1 microg/g T4 for 7 days, there was no effect on vascularized retinal area. Serum T4 levels were increased, with corresponding TSH suppression, after T4 treatment for 3 or 7 days. Serum IGF-1 levels were unaffected by T4 supplementation. CONCLUSIONS: Systemic T4 supplementation at 1 microg/g per day was detrimental to retinal vascular development in neonatal animals. If these effects are paralleled in human neonates, T4 supplementation might increase, rather than decrease, the risk of developing ROP. Further work on the role of T4 in the pathogenesis of ROP is warranted. PMID- 16354617 TI - High-resolution MAS 1H NMR spectroscopic analysis of rabbit cornea after treatment with dexamethasone and exposure to UV-B radiation. AB - Metabolic changes in rabbit cornea after combined long-term steroid treatment and UV-B exposure were investigated. Corneas were exposed to UV-B radiation (2.05 J/cm2) after 36 days topical pretreatment with either 0.1% dexamethasone or saline. Twenty-four hours after UV-B exposure, corneas were excised and aqueous humour aspirated. Intact corneal tissues were analyzed by magic angle spinning proton NMR spectroscopy and pattern recognition methods. UV-B decreased corneal ascorbate (63% reduction), taurine (62%), and choline (63%), whereas glucose was elevated. Dexamethasone pretreatment further depleted corneal taurine and ascorbate, decreased aqueous ascorbate (85%), and accumulated glucose in cornea and aqueous humour. The results reflect antioxidative mechanisms and osmoregulation. PMID- 16354618 TI - Gene transfer to trabecular meshwork endothelium via direct injection into the Schlemm canal and in vivo toxicity study. AB - PURPOSE: Our aim was to investigate the efficiency of adenoviral gene transfer via direct injection into the Schlemm canal ex vivo in human donor eyes and to examine the effect of human MMP-3 transgene expression in a rat model in vivo. METHODS: A viscocanalostomy-like operation was performed and adenoviral vector encoding for MMP-3 and green fluorescent protein was injected into human Schlemm canal or rat anterior chamber. RESULTS: Transgene expression was high in trabecular meshwork endothelium in human donor eyes. In vivo, adenovirus caused dose-dependent inflammation. CONCLUSIONS: Direct injection of adenoviral vectors into the Schlemm canal has potential in glaucoma treatment. PMID- 16354619 TI - Mast cell chymase decreases the barrier function and inhibits the migration of corneal epithelial cells. AB - PURPOSE: We investigated the in vitro effects of human mast cell chymase on corneal epithelial cells. METHODS: Human corneal epithelial cells were incubated with human chymase at activity levels that were likely to exist in the tears of patients with vernal keratoconjunctivitis. RESULTS: Incubation of chymase resulted in a decrease of barrier function of corneal epithelium. Occludin protein was cleaved by chymase. In the wound assay, incubation with chymase resulted in an inhibition of cell migration. CONCLUSION: Human chymase causes the proteolysis of occludin and fibronectin, resulting in a decrease of barrier function and inhibition of the migration of corneal epithelial cells. PMID- 16354620 TI - Dispersion of DMPC liposomes in contact lenses for ophthalmic drug delivery. AB - Approximately 90% of all ophthalmic drug formulations are now applied as eyedrops. Although eyedrops are convenient and well accepted by patients, about 95% of the drug contained in the drops is lost due to absorption through the conjunctiva or through the tear drainage. A major fraction of the drug eventually enters the bloodstream and may cause side effects. To reduce drug loss and side effects, it is proposed to encapsulate the ophthalmic drug formulations in liposomes and to disperse the drug-laden liposomes in the lens material. Upon insertion into the eye, the liposome-laden lens will slowly release the drug into the pre-lens (the film between the air and the lens) and the post-lens (the film between the cornea and the lens) tear films and thus provide drug delivery for extended periods of time. This paper focuses on dispersing dimyristoyl phosphatidylcholine (DMPC) liposomes in poly-2-hydroxyethyl methacrylate (p-HEMA) hydrogels, which are a common contact lens material. The results of this study show that the p-HEMA gels loaded with liposomes are transparent and that these gels release drugs for a period of about 8 days. Contact lenses made of particle laden gels are expected to deliver drugs at therapeutic levels for a few days. The delivery rates can be tailored by controlling the particle and the drug loading. PMID- 16354621 TI - Molecular genetic analysis of two functional candidate genes in the autosomal recessive retinitis pigmentosa, RP25, locus. AB - PURPOSE: To identify the disease gene in five Spanish families with autosomal recessive retinitis pigmentosa (arRP) linked to the RP25 locus. Two candidate genes, EEF1A1 and IMPG1, were selected from the region between D6S280 and D6S1644 markers where the families are linked. The genes were selected as good candidates on the basis of their function, tissue expression pattern, and/or genetic data. METHODS: A molecular genetic study was performed on DNA extracted from one parent and one affected member of each studied family. The coding exons, splice sites, and the 5' UTR of the genes were amplified by polymerase chain reaction (PCR). For mutation detection, direct sequence analysis was performed using the ABI 3100 automated sequencer. Segregation of an IMPG1 single nucleotide polymorphism (SNP) in all the families studied was analyzed by restriction enzyme digest of the amplified gene fragments. RESULTS: In total, 15 SNPs were identified of which 7 were novel. Of the identified SNPs, one was insertion, two were deletions, five were intronic, six were missense, and one was located in the 5' UTR. These changes, however, were also identified in unaffected members of the families and/or 50 control Caucasians. The examined known IMPG1 SNP was not segregating with the disease phenotype but was correlating with the genetic data in all families studied. CONCLUSIONS: Our results indicate that neither EEF1A1 nor IMPG1 could be responsible for RP25 in the studied families due to absence of any pathogenic variants. However, it is important to notice that the methodology used in this study cannot detect larger deletions that lie outside the screened regions or primer site mutations that exist in the heterozygous state. A role of both genes in other inherited forms of RP and/or retinal degenerations needs to be elucidated. PMID- 16354622 TI - Serotonergic retinopetal axons in the monkey retina. AB - PURPOSE: To describe serotonergic retinopetal axons in monkeys. METHODS: Whole macaque and baboon retinas, fixed in 4% paraformaldehyde, were labeled with antisera raised against serotonin (5-HT). RESULTS: Several large-diameter 5-HT immunoreactive (IR) axons emerged from the optic disk. Most axons ran to the peripheral retina, where they branched extensively. Most terminated in the ganglion cell layer, but a few 5-HT-IR axons terminated in distal inner plexiform or within inner nuclear layer. Some axons branched extensively near the fovea, and a dense plexus of 5-HT-IR axons was also found around the optic disk. Varicose 5-HT-IR axons were also associated with blood vessels, especially in the central retina. CONCLUSIONS: Immunoreactive serotonin is present in a distinct population of retinopetal axons in the monkey retina. Receptors for serotonin are present in the primate retinas, and based on physiological studies in other mammals, these retinopetal axons are expected to modulate neuronal activity and regulate blood flow. PMID- 16354623 TI - The fibrinolysis inhibitor alpha2-antiplasmin in the human cornea. AB - PURPOSE: To determine whether the cornea contains and expresses, at the gene level, the major plasmin inhibitor alpha2-antiplasmin. METHODS: Corneal sections were immunostained for alpha2-antiplasmin. Extracts of human corneal stroma, epithelium, and endothelium were subjected to immunodot blot and Western blot analysis. Total RNA and alpha2-antiplasmin specific primers were used for RT-PCR. The cDNA was sequenced. RESULTS: Alpha2-antiplasmin was observed in all three corneal layers by immunolocalization and Western blots. The major alpha2 antiplasmin form observed in most extracts was the 70-kDa form. Total alpha2 antiplasmin was present at 0.119 +/- 0.014 microg/epithelium (n = 10) and 1.45 +/ 0.47 microg/stroma (n = 10). Alpha2-antiplasmin mRNA was detected in epithelial and stromal extracts and cultured human corneal stromal cells. The sequences of the PCR products were identical to that for human alpha2-antiplasmin. CONCLUSIONS: Alpha2-antiplasmin and its mRNA are present in the cornea and may serve to regulate corneal plasmin activity. PMID- 16354624 TI - Effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on IL-6, IL 8, and MCP-1 expression in human retinal pigment epithelial cell line. AB - PURPOSE: To examine pituitary adenylate cyclase-activating polypeptide (PACAP) receptors (PAC1, VPAC1, and VPAC2) mRNA and the effect of PACAP on interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) expression in human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1beta (IL-1beta). METHODS: Expression of PACAP receptor mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR). PACAP and IL-1beta were added to serum-free medium. IL-6, IL-8, and MCP-1 mRNA were measured by real-time PCR. IL-6, IL-8, and MCP-1 protein concentrations were measured using enzyme-linked immunosorbent assay. Nuclear factor kappaB (NF kappaB) translocation was examined by immunofluorescence. RESULTS: PAC1 and VCAP1 receptors mRNA were expressed in unstimulated cells. VCAP2 mRNA was expressed in cells stimulated with IL-1beta. IL-1beta stimulated IL-6, IL-8, and MCP-1 mRNA expression and protein levels. PACAP (10(- 7) to 10(- 6) M) inhibited IL-1beta stimulated IL-6, IL-8, and MCP-1 mRNA and protein levels. Immunofluorescence of NF-kappaB in the nucleus was dense 30 min after stimulation with IL-1beta, and it was decreased by PACAP. CONCLUSIONS: ARPE-19 cells had PACAP receptors mRNA. PACAP inhibited IL-6, IL-8, and MCP-1 expression and protein secretion. Possibly, the effect on cytokines may be via suppression of NF-kappaB translocation. PMID- 16354625 TI - Fungal spectrum identified by a new slide culture and in vitro drug susceptibility using Etest in fungal keratitis. AB - PURPOSE: To investigate the spectrum of fungal species causing keratitis and to test antifungal drug susceptibility to each isolate using Etest. METHODS: Microbial cultures were performed for patients who were clinically diagnosed with fungal keratitis between September 2002 and July 2004. Modified slide culture was established to identify the fungal species of the isolates. Etest (AB BIODISK, Solna, Sweden) was applied to determine the antifungal agent susceptibility of each isolate to itraconazole, fluconazole, and amphotericin B in vitro, respectively. RESULTS: Among 73 eyes of 73 patients with clinical diagnosis of fungal keratitis, 61 strains of fungi were isolated from 61 eyes. The rate of positive culture was 81.3% of all cases. The spectrum of fungal species involved: 58 (95.1%) isolates of filamentous fungi, including the two most common genera Fusarium (n = 33, 54.1%) and Aspergillus (n = 9, 14.8%),-followed by 16 (26.2%) isolates of other genera of filamentous fungi such as Alternaria (n = 3, 4.9%), Trichophyton (n = 3, 4.9%), Curvularia (n = 2, 3.3%), Chrysosporium (n = 2, 3.3%), Acremonium (n = 2, 3.3%), and Scedosporium (n = 1, 1.6%), 1 (1.6%) yeast of Candida, as well as two (3.3%) dimorphic fungi of Blastomyces and Sporothrix isolate each. Three filamentous fungi of the isolates failed to be identified according to the information provided by slide culture. The results of Etest showed that 20 (60.6%) isolates of Fusarium were susceptible to amphotericin B, whereas all of them were resistant to itraconazole and fluconazole. All nine (100%) isolates of Aspergillus were sensitive to itraconazole, whereas four (44.4%) of them were sensitive to amphotericin B, and only two (22.2%) of them were sensitive to fluconazole. Seventeen (89.5%), 13 (68.4%), and 10 (52.6%) isolates of the remaining 19 organisms were sensitive to amphotericin B, itraconazole, and fluconazole, respectively. CONCLUSIONS: Fusarium and Aspergillus are the most frequent pathogenic organisms in causing fungal keratitis, whereas other species of fungi can also cause corneal infection. In vitro Etest for assessing antifungal drug susceptibility is a simple and practical method and may provide referential information for clinical consideration of choosing antifungal agents to treat fungal keratitis. PMID- 16354626 TI - Alpha2-adrenergic receptors are present in normal human conjunctiva. AB - PURPOSE: Despite the passage of medications, including antiglaucoma drugs, through the ocular surface, and despite the increasing relevance of neurogenic inflammation in the ocular surface, the presence of some neuroreceptors in the conjunctiva has not been ascertained. This study describes the presence of alpha2 adrenergic receptors in normal human conjunctiva. METHODS: Immunofluorescence microscopy, electrophoresis, and Western blot analyses were done in human conjunctival biopsies and rat control tissues. Antibodies against alpha2 adrenergic receptor subtypes alpha2A, alpha2B, and alpha2C were used. RESULTS: Immunoreactivity for alpha2A- and alpha2B-, but not alpha2C-adrenergic receptors was evenly distributed in epithelial cells of human conjunctiva cryosections. Immunoreactive bands were detected for the three alpha2-adrenergic receptor subtypes: a major band of 48-50 kDa and fellow bands of 65-67 kDa. CONCLUSIONS: Normal human conjunctival epithelial cells express alpha2A-, alpha2B-, and alpha2C-adrenergic receptors. Further studies to determine the functional implications of these receptors in ocular surface homeostasis are warranted. PMID- 16354627 TI - Comments on "The thickness of the tear film". PMID- 16354629 TI - Use of enophthalmic implants in the repair of orbital floor fractures. AB - INTRODUCTION: High-density porous polyethylene (Medpor) enophthalmic implants are used in patients with the sunken socket syndrome to augment the orbital volume. We have used them to improve enophthalmos and hypoglobus in the repair of large long-standing orbital floor fractures. This is the first report of enophthalmic wedge implants being used in seeing eyes to improve function and cosmesis. MATERIALS AND METHODS: Four patients had large long-standing orbital floor fractures with subsequent enlargement/expansion of the orbital volume. Indications for surgical intervention were enophthalmos (> 2 mm), hypoglobus (> 2 mm) and diplopia in primary position and downgaze. A transconjunctival approach was used to access the orbital floor and a Medpor enophthalmic wedge implant was placed postero-inferiorly to the globe and periosteum. RESULTS: Patients were followed up for a minimum of 4 to 6 months post-operatively with no complications noted. All patients achieved a good cosmetic and functional result post operatively with resolution of their enophthalmos and hypoglobus. Diplopia in primary position was corrected with only mild residual diplopia present in extreme up-gaze. CONCLUSION: We have found enophthalmic implants useful in the surgical repair of symptomatic long-standing orbital floor fractures. PMID- 16354630 TI - A rationale for therapeutic decision-making in chalazia. AB - BACKGROUND: Cytologically, a chalazion may be either a mixed-cell or a suppurating granuloma. Treatment includes incision & curettage or intralesional steroid injection. The choice of treatment is based on clinical experience. OBJECTIVE: To determine whether the cytological type of chalazion has any bearing on its response to treatment. MATERIALS AND METHODS: Prospective study of 62 clinically suspected cases of chalazia subjected to fine-needle aspiration cytology followed by either incision & curettage or intralesional steroid injection. The outcome measure was the size of the residual lesion at one week and one month post-intervention. The relationship between cytology and age, duration, size of lesion, type of treatment and response was analyzed by means of Student's t-test, the chi-square test and discriminant analysis. RESULTS: Thirty five (56.5%) patients underwent incision & curettage while 27 (43.5%) received intralesional steroid injections. Cytology revealed mixed-cell granuloma in 41 (66.1%) and suppurating granuloma in 21 (33.9%) of the lesions. Patients aged > or = 35.1 years, lesions with duration > or = 8.5 months, and lesion sizes > or = 11.4 mm (correctly classified in 66.1%, 66.7% and 71.0% of the cases, respectively) were likely to have/be suppurating granulomas. Mixed-cell granulomas responded equally well to the two therapeutic modalities, while suppurating granulomas responded significantly better to incision & curettage (p = 0.008). CONCLUSION: Mixed-cell granulomas may be treated by either of the two therapeutic modalities. However, suppurating granulomas should be treated by incision & curettage. Incision & curettage should also be the procedure of choice in patients > or = 35.1 years of age, lesions with duration > or = 8.5 months, and lesions > or = 11.4 mm in size. PMID- 16354631 TI - Onset and pattern of tear secretions in full-term neonates. AB - PURPOSE: Serial evaluations of total, basal and emotional tear secretion in full term normal neonates to determine the time after birth when these parameters attain normal adult values. METHOD: Both eyes of 102 full-term normal neonates were prospectively evaluated for tear secretions in the Department of Ophthalmology, over a period of one year. Serial recording of the Schirmer I test (total tear secretions), Basal Secretion Test, and Schirmer II test (emotional tears) was done, the first one being within 6 hours of birth. All the tests were repeated in each infant until normal adult values for each test were obtained. RESULTS: The average values of the Schirmer I test, Basal Secretion Test and Schirmer II test at birth were 23.2 (+/- 3.96) mm, 6.2 (+/- 2.15) mm and 19.2 (+/ 4.94) mm, respectively. A statistically significant correlation of birth weight but not of gestational age was found with basal secretions (p = 0.004) as well as with emotional tears (Schirmer II test, p = 0.010). At birth, 98% of infants had total tear secretion, 3.9% had basal tear secretion and 2.9% had emotional tear secretion comparable to normal adult values. All the parameters of tear secretions increased with time, so that 100% of infants had total tear secretion comparable to normal adult values within 12 hours of birth. The basal secretions took three weeks and emotional tears took four weeks to attain normal adult values in all the neonates. CONCLUSION: Total tear secretions were the earliest to reach normal adult values, followed by basal secretions and, lastly, emotional tears. Reduced basal tears at birth may predispose neonates to corneal drying during prolonged ocular examinations such as indirect ophthalmoscopy and procedures under general anesthesia. PMID- 16354632 TI - Spontaneous eye blink analysis in the normal individual. AB - PURPOSE: Spontaneous eye blink activity in the primary eye position and its relationship to age and gender were assessed using digital image processing techniques to quantify blink opening and closing time. METHODS: One hundred-and eighty healthy volunteers (90 males and 90 females), divided into the age groups 0-3, 4-12, 13-20, 21-40, 41-60 and > or = 60 years old, were evaluated prospectively. They were videotaped digitally in a standard setting and the images were transferred to a personal computer (Macintosh 400) and processed with the iMovie software. Blink opening and closing time were measured at 30 frames/second. The data were then subjected to statistical analysis. RESULTS: The closing time was significantly longer than the opening time for all ages and both genders. Elderly individuals (> or = 41 years old) and women had significantly longer closing times. CONCLUSION: Image processing techniques made possible the observation of differences in spontaneous eye blink opening and closing time in relation to age and gender. PMID- 16354633 TI - Outcome of periocular basal cell carcinoma managed by overnight paraffin section. AB - BACKGROUND: Surgical excision of periocular skin cancer allows for optimum control in terms of tumour recurrence. Although Mohs' technique gives the best outcome, it is not widely available. Processing paraffin sections is slower but histologically superior to Mohs' frozen sections. We report the results of using a standard paraffin section (non-Mohs') technique to confirm histological clearance. METHOD: A retrospective study between 1/6/95 and 1/6/99 of all consecutive patients who had excision of periocular cancer was performed. All patients had surgical excision of the tumour with a 3-mm margin. Rapid (24-hour) paraffin sections were done and reconstruction performed if histological clearance was confirmed. If tumour was still present, a further 3-mm margin was excised at the appropriate edge(s) before reconstruction took place. RESULTS: This study yielded 93 basal cell carcinomas (BCCs) of which 86 were of primary origin and 7 were recurrent tumours. The tumour characteristics were as follows: 88% nodular BCCs, 82% had a maximum surface measurement less than or equal to 10 mm, 39.8% were inner canthal and 49.5% were localised to the lower lid. In the 30 (35.4%) cases that required further excision based on the initial histological reports, tumour was seen in only four (11.4%) cases. Overall histological clearance for primary BCCs was achieved in 81/86 (94.2%) cases. There was only one late recurrence at 4 years and 5 months. Among recurrent BCCs, there was one recurrence (12.5%) and this was despite histological clearance having been reported. CONCLUSION: For primary BCCs the recurrence rate in this study was low and comparable to that following Mohs' technique. This approach, using routine pathological facilities, therefore offers a viable technique for managing periocular BCCs. PMID- 16354634 TI - A comparison of human and porcine acellular dermal tissues in the subcutaneous space of a rat model. AB - The study compared the host response to a human and a porcine acellular dermal tissue implanted in the subcutaneous space of a rat model. The human and porcine acellular grafts were surgically implanted in the subcutaneous tissue of rats (5 rats/group) and the materials were evaluated at 7, 15, 30, 60 and 180 postoperative days (PO). The histological immune response was quantified using a digital image analysis system, which evaluated the number of vessels present in the implants and in the surrounding soft tissue, the area of inflammatory cell infiltration in the grafts, the width of the capsular formation present around the tissues and the area of implants absorbed. The data were submitted to statistical analysis. Light microscopy showed mononuclear cellular infiltration, the presence of a capsular formation surrounding the grafts and the presence of vacuolar structures (optically "empty spaces") inside the implants. The image analysis comparing both materials showed significant inflammatory cells in the human graft at 15 and 30 PO, thicker capsular formation in the porcine tissue at 60 PO, increased number of vessels inside the implants and in the surrounding tissues in the porcine graft and a similar absorption pattern in both materials at 180 PO. The histological findings showed that both tissues were well-tolerated when implanted in the subcutaneous tissue of rats, allowing us to consider the porcine acellular dermal graft as a provisional alternative material for reconstructive plastic surgery. PMID- 16354635 TI - Lacrimal gland orbital lobe cysts associated with MALT lymphoma and primary Sjogren's syndrome. AB - PURPOSE: To describe, for the first time, the occurrence of a large cyst in the orbital lobe of the lacrimal gland in association with marginal zone B-cell MALT lymphoma in three female patients, two of whom also had primary Sjogren's syndrome. METHODS: Retrospective case note review. RESULTS: Three female patients, aged 35, 41 and 47 years, presented with progressive proptosis caused by an enlarging cyst within the orbital lobe of the lacrimal gland. Two of the patients had primary Sjogren's syndrome. Excision of the cyst and part of the lacrimal gland revealed marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) type within the gland or wall of the cyst in all cases. The cysts were partially lined by ductular epithelium. One patient had systemic lymphoma, was treated with chemotherapy and remains in remission one year later; the other two had disease apparently confined to the lacrimal gland and are disease-free at 9 months and at three years after local irradiation, respectively. CONCLUSION: To our knowledge, these are the first reported cases of cysts affecting the orbital lobe of the lacrimal gland in patients with B-cell MALT lymphoma of the gland. Although cystic changes have been described in other exocrine glands, lung and thymus in patients with Sjogren's syndrome, this association has also not been described in the lacrimal gland before. PMID- 16354636 TI - Lower lid epiblepharon as the presenting feature of spontaneous dural carotid cavernous fistula in an Asian patient. AB - Lower lid epiblepharon is a relatively common problem among young Asian patients. Occasionally, it may be secondary to underlying orbital pathology. We report a 33 year-old Chinese patient with a dural carotid cavernous fistula who presented with an ipsilateral lower lid epiblepharon in the absence of any other significant signs. After a recurrence following primary corrective lid surgery, she was investigated further because of mild ipsilateral proptosis. CT-scans of the orbit revealed an enlarged superior ophthalmic vein and a dural carotid cavernous fistula was confirmed on angiography. Subsequent embolisation of the fistula led to resolution of the proptosis as well as the epiblepharon. We believe that extension of orbital fat to the inferior tarsal border in Asian eyelids together with a poorly fixed anterior lamella contributes to the occurrence of epiblepharon when the Asian orbit becomes congested. The new onset of such features in the Asian patient should lead one to suspect underlying orbital pathology, even in the absence of other, more classic signs. PMID- 16354637 TI - Orbital metastasis of urinary bladder carcinoma: a clinicopathologic report and review of the literature. AB - A 53-year-old male presented with a progressive mass of the left orbit. His medical history included an invasive carcinoma of the bladder diagnosed three weeks earlier. An orbital biopsy was performed and the diagnosis was that of an orbital metastasis of urinary bladder carcinoma. The patient developed widespread metastatic disease and unfortunately died one month after the diagnosis of orbital metastasis. Orbital metastasis of urinary bladder carcinoma is associated with a poor prognosis and is more frequently observed in older people. In addition, it is five times more prevalent in men than in women. PMID- 16354638 TI - Mohs' micrographic surgery for Merkel cell carcinomas of the eyelid. AB - Merkel cell carcinoma (MCC) is a rare neuroendocrine tumour that may affect the eyelid. There is no consensus on the optimal treatment for eyelid MCC, and management remains controversial. Here we present our experience in a case of eyelid MCC treated by Mohs' micrographic surgery. Although this treatment has been advocated for general skin MCCs, there is very little experience with this technique in relation to eyelid MCCs. We suggest that Mohs' micrographic surgery may be an appropriate method of treating MCCs of the eyelid. PMID- 16354639 TI - Unmasking of ocular bobbing associated with jaw movement following inferior rectus surgery. AB - MG phenomenon refers to involuntary lid movements associated with jaw movements. We report a case unilateral vertical movement of the globe in a case of monocular elevation deficit. The above mentioned movement may be an unreported variant of oculo-trigemial synkinesis. PMID- 16354640 TI - Renal cell carcinoma metastasis masquerading as recurrent orbital haematoma. AB - A 70 year old man developed orbital haemorrhage after retrobulbar anaesthesia for cataract surgery and biopsy of a persistent lateral rectus mass suggested organising haematoma. Subsequent progression of the mass was shown, on repeated biopsy, to be due to metastatic renal cell carcinoma--a tumour recognised for its angiogenic and haemorrhagic potential. PMID- 16354641 TI - Voriconazole in the treatment of fungal osteomyelitis of the orbit in the immunocompromised host. AB - Scedosporium apiospermum is an uncommon cause of fungal infection in the orbit and sinus. Morbidity and mortality rates in immunocompromised patients are high. We review the first case of Scedosporium apiospermum orbito-sinus infection that was successfully treated with the new antifungal agent voriconazole. We highlight the efficacy of this drug, particularly in the treatment of osteomyelitis of the orbit and sinus, which may lead to an improvement in survival rates after Scedosporium infection. PMID- 16354642 TI - A case of primary adenocarcinoma of the lacrimal sac. AB - PURPOSE: To present a rare case of adenocarcinoma arising from the lacrimal sac. DESIGN: Interventional case report. METHODS: Report of a new case of primary lacrimal sac adenocarcinoma. RESULTS: The clinical presentation, workup, surgical treatment and pathology of a case of adenocarcinoma of the lacrimal sac are described. CONCLUSION: Although very rare, these tumors should be in the differential diagnosis of any lacrimal sac mass, especially those presenting above the medial canthal tendon. Other common features include expressible blood from the punctum, lack of pain, and a history of chronic epiphora or intermittent dacryocystitis. Local destruction and metastatic spread can be prevented by a high index of suspicion and early intervention. PMID- 16354643 TI - Ambulation in a wheelchair-bound stroke survivor using a walker with body weight support: a case report. AB - Treadmill systems with body weight support (BWS) have been demonstrated to increase over-ground walking speed, endurance, and balance. However, their use is limited to physical therapy gyms. Training cannot be carried over to home without significant expense. We present the case of a 43-year-old white female with no past medical history who suffered significant neurological impairment as a result of a basilar artery aneurysm. The patient was issued a walker system with BWS that allowed her to function significantly better than before she received the walker system. Implications of using the walker system with BWS in the inpatient, outpatient, and home settings are discussed. PMID- 16354644 TI - Still going, growing.....Twenty-three years after a stroke. PMID- 16354646 TI - Adult perpetrator gender asymmetries in child sexual assault victim selection: results from the 2000 National Incident-Based Report System. AB - Data from the 2000 National Incident-Based Reporting System (NIBRS) show that while males make up about nine out of every 10 adult sexual assault perpetrators, totaling about 26,878 incidents within the reporting period, females account for about one out of 10 perpetrators, totaling about 1,162 incidents. Male sexual assault perpetrators offend against child victims about 25% of the time and predominantly choose female child victims, whereas female perpetrators offend against child victims about 40% of the time and choose child victims of both genders equally. Male perpetrators offend against adolescent victims about 40% of the time, and once again tend to choose female adolescent victims. Female perpetrators offend against adolescent victims a comparable amount of time (about 45%), and for forcible offenses (rape, sodomy, sexual assault with an object, and forcible fondling) choose adolescent victims of both genders equally, while for non-forcible offenses (non-forcible incest and statutory rape) they tend to choose predominantly male victims. Finally, adult male sexual assault perpetrators choose adult victims about 36% of the time while female perpetrators choose adult victims only 16% of the time. Implications for professionals are discussed, including recommendations to aid in correct identification of adult perpetrators and child/adolescent victims of sexual assault. PMID- 16354647 TI - Chronic avoidance helps explain the relationship between severity of childhood sexual abuse and psychological distress in adulthood. AB - Recent studies have found that chronic avoidance of unpleasant internal experiences (e.g., thoughts, emotions, memories) is a maladaptive means of affect regulation often adopted by women with a history of sexual victimization in childhood. The primary aim of this study was to replicate and extend previous findings suggesting that higher levels of experiential avoidance may account for the relationship between childhood sexual abuse (CSA) and psychological distress in adulthood. It was hypothesized that, in a sample of undergraduate females (n = 151), the relationship between severity of CSA (e.g., frequency, nature of victimization) and trauma-related psychological distress would be mediated by avoidance. Results supported this hypothesis. Findings are consistent with previous studies, and further suggest that the general tendency to avoid or escape from unpleasant internal experiences may be a specific factor that exacerbates psychological distress among women with a history of sexual victimization in childhood. PMID- 16354648 TI - Cognitive distortions about sex and sexual offending: a comparison of sex offending girls, delinquent girls, and girls from the community. AB - Cognitive distortions about sexual offending were examined in 11 girls who committed sexual offenses, 12 girls who committed non-sexual criminal offenses, and 21 girls with no history of sexual of non-sexual offending. Participants responded to 12 vignettes that described sexual contact between an adolescent girl and a younger boy. The vignettes varied with respect to the sexual contact portrayed and the victim's response. Girls who had sexually offended were more likely to endorse statements reflecting the belief that the offender in a sexually aggressive vignette was not responsible for initiating the sexual contact. In addition, when the victim's response to the sexual contact was clearly negative, and the degree of contact was more serious, girls who had sexually offended demonstrated more distorted beliefs about the victim than the other two groups. Similarities and differences between the current findings and studies of distorted thinking in male sexual offenders are discussed. PMID- 16354649 TI - Efficacy of a group therapy for sexually abused adolescent girls. AB - The effects of a group therapy program for teenage girls reporting child sexual abuse were evaluated by means of a pretest/ post-test design with a control group. The psycho-educational intervention consisted of an average of 20 weekly two-hour meetings. Results of the repeated analyses of variance (ANOVA) revealed a significant improvement in youth participating in the therapy when compared with the control group on measures of post-traumatic stress, internalizing and externalizing behavior problems, coping strategies, relationship with the mother, and sense of empowerment. The findings suggest that the group therapy offered by the Centre d'Intervention en Abus Sexuels pour la Famille (CIASF) was effective in reducing symptoms for sexually abused teenagers. PMID- 16354650 TI - Against the odds: the impact of woman abuse on maternal response to disclosure of child sexual abuse. AB - Although the co-occurrence of woman abuse and child sexual abuse is high little research exists exploring the impact of woman abuse on maternal response to child sexual abuse (CSA). Findings from two qualitative studies indicate the form of woman abuse to have differential impact on maternal response. Mothers who were abused in non-physical ways, psychological, or emotional, displayed more ambivalent, less supportive responses. Mothers who were physically abused by their partners more often acted supportively to their children and displayed less ambivalence about separating from the perpetrator. Some survivors reported non disclosure as children because of their fear of the perpetrator due to the woman abuse they were exposed to while being sexually abused. Practice and research implications are discussed. PMID- 16354651 TI - Childhood physical and sexual abuse in China. AB - In order to determine the prevalence and characteristics of childhood physical and sexual abuse in China, the authors conducted a survey in Shanghai. The Dissociative Disorders Interview Schedule was administered to 423 inpatients and 304 outpatients at Shanghai Mental Health Center, and to a non-clinical sample of 618 workers at a clothing factory. The results were compared to a previous sample of 502 respondents in the general population in Winnipeg, Canada. The identities of the perpetrators of physical and sexual abuse, and the types of sexual abuse reported were similar in the two countries; however, the rates of reported abuse were lower in China. Childhood sexual abuse appears to be far less common in the general population in Canada than in China. PMID- 16354652 TI - Peptidoglycan recognition proteins are a new class of human bactericidal proteins. AB - Skin and mucous membranes come in contact with external environment and protect tissues from infections by producing antimicrobial peptides. We report that human peptidoglycan recognition proteins 3 and 4 (PGLYRP3 and PGLYRP4) are secreted as 89-115-kDa disulfide-linked homo- and heterodimers and are bactericidal against several pathogenic and nonpathogenic transient, but not normal flora, Gram positive bacteria. PGLYRP3 and PGLYRP4 are also bacteriostatic toward all other tested bacteria, which include Gram-negative bacteria and normal flora Gram positive bacteria. PGLYRP3 and PGLYRP4 are also active in vivo and protect mice against experimental lung infection. In contrast to antimicrobial peptides, PGLYRPs kill bacteria by interacting with their cell wall peptidoglycan, rather than permeabilizing their membranes. PGLYRP3 and PGLYRP4 are expressed in the skin, eyes, salivary glands, throat, tongue, esophagus, stomach, and intestine. Thus, we have identified the function of mammalian PGLYRP3 and PGLYRP4, and show that they are a new class of bactericidal and bacteriostatic proteins that have different structures, mechanism of actions, and expression patterns than antimicrobial peptides. PMID- 16354653 TI - Characterization of a counterpart to Mammalian ornithine decarboxylase antizyme in prokaryotes. AB - The degradation of mammalian ornithine decarboxylase (ODC) (EC 4.1.1.17) by 26 S proteasome, is accelerated by the ODC antizyme (AZ), a trigger protein involved in the specific degradation of eukaryotic ODC. In prokaryotes, AZ has not been found. Previously, we found that in Selenomonas ruminantium, a strictly anaerobic and Gram-negative bacterium, a drastic degradation of lysine decarboxylase (LDC; EC 4.1.1.18), which has decarboxylase activities toward both L-lysine and L ornithine with similar K(m) values, occurs upon entry into the stationary phase of cell growth by protease together with a protein of 22 kDa (P22). Here, we show that P22 is a direct counterpart of eukaryotic AZ by the following evidence. (i) P22 synthesis is induced by putrescine but not cadaverine. (ii) P22 enhances the degradation of both mouse ODC and S. ruminantium LDC by a 26 S proteasome. (iii) S. ruminantium LDC degradation is also enhanced by mouse AZ replacing P22 in a cell-free extract from S. ruminantium. (iv) Both P22 and mouse AZ bind to S. ruminantium LDC but not to the LDC mutated in its binding site for P22 and AZ. In this report, we also show that P22 is a ribosomal protein of S. ruminantium. PMID- 16354654 TI - Cathepsin D is present in human eccrine sweat and involved in the postsecretory processing of the antimicrobial peptide DCD-1L. AB - The protein pattern of healthy human eccrine sweat was investigated and 10 major proteins were detected from which apolipoprotein D, lipophilin B, and cathepsin D (CatD) were identified for the first time in human eccrine sweat. We focused our studies on the function of the aspartate protease CatD in sweat. In vitro digestion experiments using a specific fluorescent CatD substrate showed that CatD is enzymatically active in human sweat. To identify potential substrates of CatD in human eccrine sweat LL-37 and DCD-1L, two antimicrobial peptides present in sweat, were digested in vitro with purified CatD. LL-37 was not significantly digested by CatD, whereas DCD-1L was cleaved between Leu(44) and Asp(45) and between Leu(29) and Glu(30) almost completely. The DCD-1L-derived peptides generated in vitro by CatD were also found in vivo in human sweat as determined by surface-enhanced laser desorption/ionization (SELDI) mass spectrometry. Furthermore, besides the CatD-processed peptides we identified additionally DCD 1L-derived peptides that are generated upon cleavage with a 1,10-phenanthroline sensitive carboxypeptidase and an endoprotease. Taken together, proteolytic processing generates 12 DCD-1L-derived peptides. To elucidate the functional significance of postsecretory processing the antimicrobial activity of three CatD processed DCD-1L peptides was tested. Whereas two of these peptides showed no activity against Gram-positive and Gram-negative bacteria, one DCD-1L-derived peptide showed an even higher activity against Escherichia coli than DCD-1L. Functional analysis indicated that proteolytic processing of DCD-1L by CatD in human sweat modulates the innate immune defense of human skin. PMID- 16354655 TI - A novel thioredoxin h is secreted in Nicotiana alata and reduces S-RNase in vitro. AB - Thioredoxins type h are classified into three subgroups. The subgroup II includes thioredoxins containing an N-terminal extension, the role of which is still unclear. Although thioredoxin secretion has been observed in animal cells, there is no evidence suggesting that any thioredoxin h is secreted in plants. In this study, we report that a thioredoxin h, subgroup II, from Nicotiana alata (NaTrxh) is secreted into the extracellular matrix of the stylar transmitting tract tissue. Fractionation studies showed that NaTrxh is extracted along with well characterized secretion proteins such as S-RNases and NaTTS (N. alata transmitting tissue-specific protein). Moreover, an NaTrxh-green fluorescent fusion protein transiently expressed in Nicotiana benthamiana and Arabidopsis thaliana leaves was also secreted, showing that NaTrxh has the required information for its secretion. We performed reduction assays in vitro to identify potential extracellular targets of NaTrxh. We found that S-RNase is one of the several potential substrates of the NaTrxh in the extracellular matrix. In addition, we proved by affinity chromatography that NaTrxh specifically interacts with S-RNase. Our findings showed that NaTrxh is a new thioredoxin h in Nicotiana that is secreted as well as in animal systems. Because NaTrxh is localized in the extracellular matrix of the stylar transmitting tract and its specific interaction with S-RNase to reduce it in vitro, we suggest that this thioredoxin h may be involved either in general pollen-pistil interaction processes or particularly in S-RNase-based self-incompatibility. PMID- 16354656 TI - Stabilization of a stalled replication fork by concerted actions of two helicases. AB - PriA helicase plays crucial roles in restoration of arrested replication forks. It carries a "3' terminus binding pocket" in its N-terminal DNA binding domain, which is required for high affinity binding of PriA to a fork carrying a 3'-end of a nascent leading strand at the branch. We show that the abrogation of the 3' terminus recognition either by a mutation in the 3' terminus binding pocket or by the bulky modification of the 3'-end leads to unwinding of the unreplicated duplex arm on this fork, causing potential fork destabilization. This indicates a critical role of the 3' terminus binding pocket of PriA in its "stable" binding at the fork for primosome assembly. In contrast, PriA unwinds the unreplicated duplex region on a fork without a 3'-end, potentially destabilizing the fork. However, this process is inhibited by RecG helicase, capable of regressing the fork until the 3'-end of the nascent leading strand reaches the branch. PriA now stably binds to this regressed fork, stabilizing it. Using a model arrest-fork substrate, we reconstitute the above process in vitro with RecG and PriA proteins. Our results present a novel mechanism by which two helicases function in a highly coordinated manner to generate a structure in which an arrested fork is stabilized for further repair and/or replication restart. PMID- 16354657 TI - Compensatory increase in hepatic lipogenesis in mice with conditional intestine specific Mttp deficiency. AB - Microsomal TG transfer protein (MTTP) is required for the assembly and secretion of TG (TG)-rich lipoproteins from both enterocytes and hepatocytes. Liver specific deletion of Mttp produced a dramatic reduction in plasma very low density lipoprotein-TG and virtually eliminated apolipoprotein B100 (apoB100) secretion yet caused only modest reductions in plasma apoB48 and apoB48 secretion from primary hepatocytes. These observations prompted us to examine the phenotype following intestine-specific Mttp deletion because murine, like human enterocytes, secrete virtually exclusively apoB48. We generated mice with conditional Mttp deletion in villus enterocytes (Mttp-IKO), using a tamoxifen inducible, intestine-specific Cre transgene. Villus enterocytes from chow-fed Mttp-IKO mice contained large cytoplasmic TG droplets and no chylomicron-sized particles within the secretory pathway. Chow-fed, Mttp-IKO mice manifested steatorrhea, growth arrest, and decreased cholesterol absorption, features that collectively recapitulate the phenotype associated with abetalipoproteinemia. Chylomicron secretion was reduced dramatically in vivo, in conjunction with an approximately 80% decrease in apoB48 secretion from primary enterocytes. Additionally, although plasma and hepatic cholesterol and TG content were decreased, Mttp-IKO mice demonstrated a paradoxical increase in both hepatic lipogenesis and very low density lipoprotein secretion. These findings establish distinctive features for MTTP involvement in intestinal chylomicron assembly and secretion and suggest that hepatic lipogenesis undergoes compensatory induction in the face of defective intestinal TG secretion. PMID- 16354658 TI - A novel principle for partial agonism of liver X receptor ligands. Competitive recruitment of activators and repressors. AB - Partial, selective activation of nuclear receptors is a central issue in molecular endocrinology but only partly understood. Using LXRs as an example, we show here that purely agonistic ligands can be clearly and quantitatively differentiated from partial agonists by the cofactor interactions they induce. Although a pure agonist induces a conformation that is incompatible with the binding of repressors, partial agonists such as GW3965 induce a state where the interaction not only with coactivators, but also corepressors is clearly enhanced over the unliganded state. The activities of the natural ligand 22(R) hydroxycholesterol and of a novel quinazolinone ligand, LN6500 can be further differentiated from GW3965 and T0901317 by their weaker induction of coactivator binding. Using biochemical and cell-based assays, we show that the natural ligand of LXR is a comparably weak partial agonist. As predicted, we find that a change in the coactivator to corepressor ratio in the cell will affect NCoR recruiting compounds more dramatically than NCoR-dissociating compounds. Our data show how competitive binding of coactivators and corepressors can explain the tissue specific behavior of partial agonists and open up new routes to a rational design of partial agonists for LXRs. PMID- 16354659 TI - Light-induced oxidation of photoreceptor outer segment phospholipids generates ligands for CD36-mediated phagocytosis by retinal pigment epithelium: a potential mechanism for modulating outer segment phagocytosis under oxidant stress conditions. AB - Clearance by the retinal pigment epithelium (RPE) of shed photoreceptor outer segments (OSs), a tissue with one of the highest turnover rates in the body, is critical to the maintenance and normal function of the retina. We hypothesized that there is a potential role for photo-oxidation in OS uptake by RPE via scavenger receptor-mediated recognition of structurally defined lipid peroxidation products. We now demonstrate that specific structurally defined oxidized species derived from arachidonyl, linoleoyl, and docosahexanoyl phosphatidylcholine may serve as endogenous ligands on OSs for uptake by RPE via the scavenger receptor CD36. Mass spectrometry studies of retinal lipids recovered from dark-adapted rats following physiological light exposure demonstrate in vivo formation of specific oxidized phosphatidylcholine molecular species possessing a CD36 recognition motif, an oxidatively truncated sn-2 acyl group with a terminal gamma-hydroxy(or oxo)-alpha,beta-unsaturated carbonyl. Cellular studies using RPE isolated from wild-type versus CD36 null mice suggest that CD36 plays a role in engulfment, but not initial binding, of OSs via these oxidized phospholipids. Parallel increases in OS protein-bound nitrotyrosine, a post-translational modification by nitric oxide (NO)-derived oxidants, were also observed, suggesting a possible role for light-induced generation of NO-derived oxidants in the initiation of OS lipid peroxidation. Collectively, these studies suggest that intense light exposure promotes "oxidative tagging" of photoreceptor outer segments with structurally defined choline glycerophospholipids that may serve as a physiological signal for CD36-mediated phagocytosis under oxidant stress conditions. PMID- 16354660 TI - Role of the PAR1 receptor 8th helix in signaling: the 7-8-1 receptor activation mechanism. AB - The protease-activated receptors are tethered ligand G protein-coupled receptors that are activated by proteolytic cleavage of the extracellular domain of the receptor. The archetypic protease-activated receptor PAR1 strongly activates G(q) signaling pathways, but very little is known regarding the mechanism of signal transference between receptor and internally located G protein. The recent x-ray structure of rhodopsin revealed the presence of a highly conserved amphipathic 8th helix that is likely to be physically interposed between receptor and G protein. We found that the analogous 8th helix region of PAR1 was critical for activation of G(q)-dependent signaling. Engineering an 8th helix alpha-aneurysm with a downwards-directed alanine residue markedly interfered with signal transference to G(q). The 8th helix-anchoring cysteine palmitoylation sites were important for the affinity of ligand-dependent G protein coupling but did not affect the maximal signal. A network of H-bond and ionic interactions was found to connect the N-terminal portion of the 8th helix to the nearby NPXXY motif on transmembrane helix 7 and also to the adjacent intracellular loop-1. Disruption of these pairwise interactions caused additive defects in coupling to G protein, indicating that the transmembrane 7-8th helix-i1 loop may move in a coordinated manner to transfer the signal from PAR1 to G protein. This "7-8-1" interaction network was found to be prevalent in G protein-coupled receptors involved in endothelial signaling and angiogenesis. PMID- 16354661 TI - A novel lipase belonging to the hormone-sensitive lipase family induced under starvation to utilize stored triacylglycerol in Mycobacterium tuberculosis. AB - Twenty-four putative lipase/esterase genes of Mycobacterium tuberculosis H37Rv were expressed in Escherichia coli and assayed for long-chain triacylglycerol (TG) hydrolase activity. We show here that the product of Rv3097c (LIPY) hydrolyzed long-chain TG with high specific activity. LIPY was purified after solubilization from inclusion bodies; the enzyme displayed a K(m) of 7.57 mM and V(max) of 653.3 nmol/mg/min for triolein with optimal activity between pH 8.0 and pH 9.0. LIPY was inhibited by active serine-directed reagents and was inactivated at temperatures above 37 degrees C. Detergents above their critical micellar concentrations and divalent cations inhibited the activity of LIPY. The N terminal half of LIPY showed sequence homology with the proline glutamic acid polymorphic GC-rich repetitive sequences protein family of M. tuberculosis. The C terminal half of LIPY possesses amino acid domains homologous with the hormone sensitive lipase family and the conserved active-site motif GDSAG. LIPY shows low sequence identity with the annotated lipases of M. tuberculosis and with other bacterial lipases. We demonstrate that hypoxic cultures of M. tuberculosis, which had accumulated TG, hydrolyzed the stored TG when subjected to nutrient starvation. Under such conditions, lipY was induced more than all lipases, suggesting a central role for it in the utilization of stored TG. We also show that in the lipY-deficient mutant, TG utilization was drastically decreased under nutrient-deprived condition. Thus, LIPY may be responsible for the utilization of stored TG during dormancy and reactivation of the pathogen. PMID- 16354662 TI - Interactions of nitroaromatic compounds with the mammalian selenoprotein thioredoxin reductase and the relation to induction of apoptosis in human cancer cells. AB - Here we described novel interactions of the mammalian selenoprotein thioredoxin reductase (TrxR) with nitroaromatic environmental pollutants and drugs. We found that TrxR could catalyze nitroreductase reactions with either one- or two electron reduction, using its selenocysteine-containing active site and another redox active center, presumably the FAD. Tetryl and p-dinitrobenzene were the most efficient nitroaromatic substrates with a k(cat) of 1.8 and 2.8 s(-1), respectively, at pH 7.0 and 25 degrees C using 50 muM NADPH. As a nitroreductase, TrxR cycled between four- and two-electron-reduced states. The one-electron reactions led to superoxide formation as detected by cytochrome c reduction and, interestingly, reductive N-denitration of tetryl or 2,4-dinitrophenyl-N methylnitramine, resulting in the release of nitrite. Most nitroaromatics were uncompetitive and noncompetitive inhibitors with regard to NADPH and the disulfide substrate 5,5'-dithiobis(2-nitrobenzoic acid), respectively. Tetryl and 4,6-dinitrobenzofuroxan were, however, competitive inhibitors with respect to 5,5'-dithiobis(2-nitrobenzoic acid) and were clearly substrates for the selenolthiol motif of the enzyme. Furthermore, tetryl and 4,6-dinitrobenzofuroxan efficiently inactivated TrxR, likely by alkylation of the selenolthiol motif as in the inhibition of TrxR by 1-chloro-2,4-dinitrobenzene/dinitrochlorobenzene (DNCB) or juglone. The latter compounds were the most efficient inhibitors of TrxR activity in a cellular context. DNCB, juglone, and tetryl were highly cytotoxic and induced caspase-3/7 activation in HeLa cells. Furthermore, DNCB and juglone were potent inducers of apoptosis also in Bcl2 overexpressing HeLa cells or in A549 cells. Based on these findings, we suggested that targeting of intracellular TrxR by alkylating nitroaromatic or quinone compounds may contribute to the induction of apoptosis in exposed human cancer cells. PMID- 16354663 TI - Identification and characterization of Rv3281 as a novel subunit of a biotin dependent acyl-CoA Carboxylase in Mycobacterium tuberculosis H37Rv. AB - Mycobacterium tuberculosis produces a large number of structurally diverse lipids generated from the carboxylation products of acetyl-CoA and propionyl-CoA. A biotin-dependent acyl-CoA carboxylase was purified from M. tuberculosis H37Rv by avidin affinity chromatography, and the three major protein components were determined by N-terminal sequencing to be the 63-kDa alpha3-subunit (AccA3, Rv3285), the 59-kDa beta5-subunit (AccD5, Rv3280), and the 56-kDa beta4-subunit (AccD4, Rv3799). A minor protein of about 24 kDa that co-purified with the above subunits was identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry to be the product of Rv3281 that is located immediately downstream of the open reading frame encoding the beta5-subunit. This protein displays identity over a short stretch of amino acids with the recently discovered epsilon-subunits of Streptomyces coelicolor, suggesting that it might be an epsilon-subunit of the mycobacterial acyl-CoA carboxylase. To test this hypothesis, the carboxylase subunits were expressed in Escherichia coli and purified. Acyl-CoA carboxylase activity was successfully reconstituted for the first time from purified subunits of the acyl-CoA carboxylase of M. tuberculosis. The reconstituted alpha3-beta5 showed higher activity with propionyl-CoA than with acetyl-CoA, and the addition of the epsilon-subunit stimulated the carboxylation by 3.2- and 6.3-fold, respectively. The alpha3-beta4 showed very low activity with the above substrates but carboxylated long chain acyl-CoA. This epsilon-subunit contains five sets of tandem repeats at the N terminus that are required for maximal enhancement of carboxylase activity. The Rv3281 open reading frame is co-transcribed with Rv3280 in the mycobacterial cell, and the level of epsilon-protein was highest during the log phase and decreased during the stationary phase. PMID- 16354664 TI - Small weak acids reactivate proton transfer in reaction centers from Rhodobacter sphaeroides mutated at AspL210 and AspM17. AB - In reaction centers of Rhodobacter sphaeroides, site-directed mutagenesis has implicated several acidic residues in the delivery of protons to the secondary quinone (Q(B)) during reduction to quinol. In a double mutant (Asp(L210) --> Asn + Asp(M17) --> Asn) that is severely impaired in proton transfer capability over a wide pH range, proton transfer was "rescued" by added weak acids. For low pK(a) acids the total concentration of salt required near neutral pH was high. The ionic strength effect of added salts stimulated the rate of proton-coupled electron transfer at pH < 7, but decreased it at pH > 7.5, indicating an effective isoelectric point between these limits. In this region, a substantial rate enhancement by weak acids was clearly evident. A Bronsted plot of activity versus pK(a) of the rescuing acids was linear, with a slope of -1, and extrapolated to a diffusion-limited rate at pK(a)(app) approximately 1. However, the maximum rate at saturating concentrations of acid did not correlate with pK(a), indicating that the acid and anion species compete for binding, both with weak affinity. This model predicts that pK(a)(app) corresponds to a true pK(a) = 4-5, similar to that for a carboxylic acid or Q(B)(-), itself. Only rather small, neutral acids were active, indicating a need to access a small internal volume, suggested to be a proton channel to the Q(B) domain. However, the on-rates were near the diffusion limit. The implications for intraprotein proton transfer pathway design are discussed. PMID- 16354665 TI - Release of the soluble transferrin receptor is directly regulated by binding of its ligand ferritransferrin. AB - The human transferrin receptor (TfR) is shed by an integral metalloprotease releasing a soluble form (sTfR) into serum. The sTfR reflects the iron demand of the body and is postulated as a regulator of iron homeostasis via binding to the hereditary hemochromatosis protein HFE. To study the role of transferrin in this process, we investigated TfR shedding in HL60 cells and TfR-deficient Chinese hamster ovary cells transfected with human TfR. Independent of TfR expression, sTfR release decreases with increasing ferritransferrin concentrations, whereas apo-transferrin exhibits no inhibitory effect. To investigate the underlying mechanism, we generated several TfR mutants with different binding affinities for transferrin. Shedding of TfR mutants in transfected cells correlates exactly with their binding affinity, implying that the effect of ferritransferrin on TfR shedding is mediated by a direct molecular interaction. Analysis of sTfR release from purified microsomal membranes revealed that the regulation is independent from intracellular trafficking or cellular signaling events. Our results clearly demonstrated that sTfR does not only reflect the iron demand of the cells but also the iron availability in the bloodstream, mirrored by iron saturation of transferrin, corroborating the important potential function of sTfR as a regulator of iron homeostasis. PMID- 16354666 TI - Aminoglycosides decrease glutathione peroxidase-1 activity by interfering with selenocysteine incorporation. AB - Cellular glutathione peroxidase is a key intracellular antioxidant enzyme that contains a selenocysteine residue at its active site. Selenium, a selenocysteine incorporation sequence in the 3'-untranslated region of the glutathione peroxidase mRNA, and other translational cofactors are necessary for "read through" of a UGA stop codon that specifies selenocysteine incorporation. Aminoglycoside antibiotics facilitate read-through of premature stop codons in prokayotes and eukaryotes. We studied the effects of G418, an aminoglycoside, on cellular glutathione peroxidase expression and function in mammalian cells. Insertion of a selenocysteine incorporation element along with a UGA codon into a reporter construct allows for read-through only in the presence of selenium. G418 increased read-through in selenium-replete cells as well as in the absence of selenium. G418 treatment increased immunodetectable endogenous or recombinant glutathione peroxidase but reduced the specific activity of the enzyme. Tandem mass spectrometry experiments indicated that G418 caused a substitution of l arginine for selenocysteine. These data show that G418 can affect the biosynthesis of this key antioxidant enzyme by promoting substitution at the UGA codon. PMID- 16354667 TI - Rational design of intercellular adhesion molecule-1 (ICAM-1) variants for antagonizing integrin lymphocyte function-associated antigen-1-dependent adhesion. AB - The interaction between integrin lymphocyte function-associated antigen-1 (LFA-1) and its ligand intercellular adhesion molecule-1 (ICAM-1) is critical in immunological and inflammatory reactions but, like other adhesive interactions, is of low affinity. Here, multiple rational design methods were used to engineer ICAM-1 mutants with enhanced affinity for LFA-1. Five amino acid substitutions 1) enhance the hydrophobicity and packing of residues surrounding Glu-34 of ICAM-1, which coordinates to a Mg2+ in the LFA-1 I domain, and 2) alter associations at the edges of the binding interface. The affinity of the most improved ICAM-1 mutant for intermediate- and high-affinity LFA-1 I domains was increased by 19 fold and 22-fold, respectively, relative to wild type. Moreover, potency was similarly enhanced for inhibition of LFA-1-dependent ligand binding and cell adhesion. Thus, rational design can be used to engineer novel adhesion molecules with high monomeric affinity; furthermore, the ICAM-1 mutant holds promise for targeting LFA-1-ICAM-1 interaction for biological studies and therapeutic purposes. PMID- 16354668 TI - Nucleotide-binding sites in the voltage-dependent anion channel: characterization and localization. AB - In this study, we addressed the presence and location of nucleotide-binding sites in the voltage-dependent anion channel (VDAC). VDAC bound to reactive red 120 agarose, from which it was eluted by ATP, less effectively by ADP and AMP, but not by NADH. The photoreactive ATP analog, benzoyl-benzoyl-ATP (BzATP), was used to identify and characterize the ATP-binding sites in VDAC. [alpha-(32)P]BzATP bound to purified VDAC at two or more binding sites with apparent high and low binding affinities. Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) analysis of BzATP-labeled VDAC confirmed the binding of at least two BzATP molecules to VDAC. The VDAC BzATP-binding sites showed higher specificity for purine than for pyrimidine nucleotides and higher affinity for negatively charged nucleotide species. VDAC treatment with the lysyl residue modifying reagent, fluorescein 5'-isothiocyanate, markedly inhibited VDAC labeling with BzATP. The VDAC nucleotide-binding sites were localized using chemical and enzymatic cleavage. Digestion of [alpha-(32)P]BzATP-labeled VDAC with CNBr or V8 protease resulted in the appearance of approximately 17- and approximately 14-kDa labeled fragments. Further digestion, high performance liquid chromatography separation, and sequencing of the selected V8 peptides suggested that the labeled fragments originated from two different regions of the VDAC molecule. MALDI-TOF analysis of BzATP-labeled, tryptic VDAC fragments indicated and localized three nucleotide binding sites, two of which were at the N and C termini of VDAC. Thus, the presence of two or more nucleotide-binding sites in VDAC is suggested, and their possible function in the control of VDAC activity, and, thereby, of outer mitochondrial membrane permeability is discussed. PMID- 16354669 TI - Guide molecule-driven stereospecific degradation of alpha-methylpolyamines by polyamine oxidase. AB - FAD-dependent polyamine oxidase (PAO; EC 1.5.3.11) is one of the key enzymes in the catabolism of polyamines spermidine and spermine. The natural substrates for the enzyme are N1-acetylspermidine, N1-acetylspermine, and N1,N12 diacetylspermine. Here we report that PAO, which normally metabolizes achiral substrates, oxidized (R)-isomer of 1-amino-8-acetamido-5-azanonane and N1 acetylspermidine as efficiently while (S)-1-amino-8-acetamido-5-azanonane was a much less preferred substrate. It has been shown that in the presence of certain aldehydes, the substrate specificity of PAO and the kinetics of the reaction are changed to favor spermine and spermidine as substrates. Therefore, we examined the effect of several aldehydes on the ability of PAO to oxidize different enantiomers of alpha-methylated polyamines. PAO supplemented with benzaldehyde predominantly catalyzed the cleavage of (R)-isomer of alpha-methylspermidine, whereas in the presence of pyridoxal the (S)-alpha-methylspermidine was preferred. PAO displayed the same stereospecificity with both singly and doubly alpha-methylated spermine derivatives when supplemented with the same aldehydes. Structurally related ketones proved to be ineffective. This is the first time that the stereospecificity of FAD-dependent oxidase has been successfully regulated by changing the supplementary aldehyde. These findings might facilitate the chemical regulation of stereospecificity of the enzymes. PMID- 16354670 TI - Involvement of a novel Q-SNARE, D12, in quality control of the endomembrane system. AB - The cellular endomembrane system requires the proper kinetic balance of synthesis and degradation of its individual components, which is maintained in part by a specific membrane fusion apparatus. In this study, we describe the molecular properties of D12, which was identified from a mouse expression library. This C terminal anchored membrane protein has sequence similarity to both a yeast soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor (SNARE), Use1p/Slt1p, and a recently identified human syntaxin 18-binding protein, p31. D12 formed a tight complex with syntaxin 18 as well as Sec22b and bound to alpha-SNAP, indicating that D12 is a SNARE protein. Although the majority of D12 is located in the endoplasmic reticulum and endoplasmic reticulum Golgi intermediate compartments at steady state, overexpression or knockdown of D12 had no obvious effects on membrane trafficking in the early secretory pathway. However, suppression of D12 expression caused rapid appearance of lipofuscin granules, accompanied by apoptotic cell death without the apparent activation of the unfolded protein response. The typical cause of lipofuscin formation is the impaired degradation of mitochondria by lysosomal degradative enzymes, and, consistent with this, we found that proper post-Golgi maturation of cathepsin D was impaired in D12-deficient cells. This unexpected observation was supported by evidence that D12 associates with VAMP7, a SNARE in the endosomal lysosomal pathway. Hence, we suggest that D12 participates in the degradative function of lysosomes. PMID- 16354671 TI - Roles of the hydrophobic cavity and lid of LolA in the lipoprotein transfer reaction in Escherichia coli. AB - LolA, a periplasmic chaperone, binds to outer membrane-specific lipoproteins released from the inner membrane through the action of an ATP-binding cassette transporter, LolCDE and then transfers them to the outer membrane receptor LolB, thereby mediating the inner to outer membrane transport of lipoproteins. The crystal structure of free LolA revealed that it has an internal hydrophobic cavity, which is surrounded by hydrophobic residues and closed by a lid comprising alpha-helices. The hydrophobic cavity most likely represents the binding site for the lipid moiety of a lipoprotein. It is speculated that the lid undergoes opening and closing upon the binding and transfer of lipoproteins, respectively. To determine the functions of the hydrophobic cavity and lid in detail, 14 residues involved in the formation of these structures were subjected to random mutagenesis. Among the obtained 21 LolA derivatives that did not support growth, 14 were active as to the binding of lipoproteins but defective in the transfer of lipoproteins to LolB, causing the periplasmic accumulation of a lipoprotein as a complex with a LolA derivative. A LolA derivative, I93G, bound lipoproteins faster than wild-type LolA did, whereas it did not transfer associated lipoproteins to LolB. When I93G and wild type LolA co-existed, lipoproteins were bound only to I93G; which therefore exhibited a dominant negative property. Another derivative, L59R, was also defective in the transfer of lipoproteins to LolB but did not exhibit a dominant negative property. Taken together, these results indicate that both the hydrophobic cavity and the lid are critically important for not only the binding of lipoproteins but also their transfer. PMID- 16354672 TI - The holo-form of the nucleotide binding domain of the KdpFABC complex from Escherichia coli reveals a new binding mode. AB - P-type ATPases are ubiquitously abundant enzymes involved in active transport of charged residues across biological membranes. The KdpB subunit of the prokaryotic Kdp-ATPase (KdpFABC complex) shares characteristic regions of homology with class II-IV P-type ATPases and has been shown previously to be misgrouped as a class IA P-type ATPase. Here, we present the NMR structure of the AMP-PNP-bound nucleotide binding domain KdpBN of the Escherichia coli Kdp-ATPase at high resolution. The aromatic moiety of the nucleotide is clipped into the binding pocket by Phe(377) and Lys(395) via a pi-pi stacking and a cation-pi interaction, respectively. Charged residues at the outer rim of the binding pocket (Arg(317), Arg(382), Asp(399), and Glu(348)) stabilize and direct the triphosphate group via electrostatic attraction and repulsion toward the phosphorylation domain. The nucleotide binding mode was corroborated by the replacement of critical residues. The conservative mutation F377Y produced a high residual nucleotide binding capacity, whereas replacement by alanine resulted in low nucleotide binding capacities and a considerable loss of ATPase activity. Similarly, mutation K395A resulted in loss of ATPase activity and nucleotide binding affinity, even though the protein was properly folded. We present a schematic model of the nucleotide binding mode that allows for both high selectivity and a low nucleotide binding constant, necessary for the fast and effective turnover rate realized in the reaction cycle of the Kdp-ATPase. PMID- 16354673 TI - Decreased renal organic anion secretion and plasma accumulation of endogenous organic anions in OAT1 knock-out mice. AB - The "classical" organic anion secretory pathway of the renal proximal tubule is critical for the renal excretion of the prototypic organic anion, para aminohippurate, as well as of a large number of commonly prescribed drugs among other significant substrates. Organic anion transporter 1 (OAT1), originally identified as NKT (Lopez-Nieto, C. E., You, G., Bush, K. T., Barros, E. J. G., Beier, D. R., and Nigam, S. K. (1997) J. Biol. Chem. 272, 6471-6478), has physiological properties consistent with a role in this pathway. However, several other transporters (e.g. OAT2, OAT3, and MRP1) have also been proposed as important PAH transporters on the basis of in vitro studies; therefore, the relative contribution of OAT1 has remained unclear. We have now generated a colony of OAT1 knock-out mice, permitting elucidation of the role of OAT1 in the context of these other potentially functionally redundant transporters. We find that the knock-out mice manifest a profound loss of organic anion transport (e.g. para-aminohippurate) both ex vivo (in isolated renal slices) as well as in vivo (as indicated by loss of renal secretion). In the case of the organic anion, furosemide, loss of renal secretion in the knock-out results in impaired diuretic responsiveness to this drug. These results indicate a critical role for OAT1 in the functioning of the classical pathway. In addition, we have determined the levels of approximately 60 endogenous organic anions in the plasma and urine of wild-type and knock-out mice. This has led to identification of several compounds with significantly higher plasma concentrations and/or lower urinary concentrations in knock-out mice, suggesting the involvement of OAT1 in their renal secretion. We have also demonstrated in xenopus oocytes that some of these compounds interact with OAT1 in vitro. Thus, these latter compounds might represent physiological substrates of OAT1. PMID- 16354674 TI - Helicobacter pylori thioredoxin is an arginase chaperone and guardian against oxidative and nitrosative stresses. AB - The gastric human pathogen Helicobacter pylori faces formidable challenges in the stomach including reactive oxygen and nitrogen intermediates. Here we demonstrate that arginase activity, which inhibits host nitric oxide production, is post translationally stimulated by H. pylori thioredoxin (Trx) 1 but not the homologous Trx2. Trx1 has chaperone activity that renatures urea- or heat denatured arginase back to the catalytically active state. Most reactive oxygen and nitrogen intermediates inhibit arginase activity; this damage is reversed by Trx1, but not Trx2. Trx1 and arginase equip H. pylori with a "renox guardian" to overcome abundant nitrosative and oxidative stresses encountered during the persistence of the bacterium in the hostile gastric environment. PMID- 16354676 TI - Reconstitution of the Reelin signaling pathway in fibroblasts demonstrates that Dab1 phosphorylation is independent of receptor localization in lipid rafts. AB - The Reelin signaling pathway operates in migrating neurons and is indispensable for their correct positioning during embryonic brain development. Many biochemical and cell biological studies to dissect the Reelin pathway at the molecular level are hampered by the lack of a cell line harboring a functional Reelin signaling pathway. Here we present fibroblast cell lines in which all required functional components of the pathway have been reconstituted. These cells react upon Reelin treatment in the same way as primary neurons. We have subsequently used these cell lines to study the subcellular localization of ApoER2 and the VLDL receptor and could demonstrate that receptor-mediated Dab1 phosphorylation does not depend on lipid rafts and that phosphorylated Dab1 remains bound to the receptor tail when the pathway is activated by Reelin. PMID- 16354677 TI - Inhibition of SIRT1 catalytic activity increases p53 acetylation but does not alter cell survival following DNA damage. AB - Human SIRT1 is an enzyme that deacetylates the p53 tumor suppressor protein and has been suggested to modulate p53-dependent functions including DNA damage induced cell death. In this report, we used EX-527, a novel, potent, and specific small-molecule inhibitor of SIRT1 catalytic activity to examine the role of SIRT1 in p53 acetylation and cell survival after DNA damage. Treatment with EX-527 dramatically increased acetylation at lysine 382 of p53 after different types of DNA damage in primary human mammary epithelial cells and several cell lines. Significantly, inhibition of SIRT1 catalytic activity by EX-527 had no effect on cell growth, viability, or p53-controlled gene expression in cells treated with etoposide. Acetyl-p53 was also increased by the histone deacetylase (HDAC) class I/II inhibitor trichostatin A (TSA). EX-527 and TSA acted synergistically to increase acetyl-p53 levels, confirming that p53 acetylation is regulated by both SIRT1 and HDACs. While TSA alone reduced cell survival after DNA damage, the combination of EX-527 and TSA had no further effect on cell viability and growth. These results show that, although SIRT1 deacetylates p53, this does not play a role in cell survival following DNA damage in certain cell lines and primary human mammary epithelial cells. PMID- 16354678 TI - Recruitment of DNA damage checkpoint proteins to damage in transcribed and nontranscribed sequences. AB - We developed a chromatin immunoprecipitation method for analyzing the binding of repair and checkpoint proteins to DNA base lesions in any region of the human genome. Using this method, we investigated the recruitment of DNA damage checkpoint proteins RPA, Rad9, and ATR to base damage induced by UV and acetoxyacetylaminofluorene in transcribed and nontranscribed regions in wild-type and excision repair-deficient human cells in G1 and S phases of the cell cycle. We find that all 3 damage sensors tested assemble at the site or in the vicinity of damage in the absence of DNA replication or repair and that transcription enhances recruitment of checkpoint proteins to the damage site. Furthermore, we find that UV irradiation of human cells defective in excision repair leads to phosphorylation of Chk1 kinase in both G1 and S phase of the cell cycle, suggesting that primary DNA lesions as well as stalled transcription complexes may act as signals to initiate the DNA damage checkpoint response. PMID- 16354679 TI - Transactivation of platelet-derived growth factor receptor alpha by the GTPase deficient activated mutant of Galpha12. AB - The GTPase-deficient, activated mutant of Galpha12 (Galpha12Q229L, or Galpha12QL) induces neoplastic growth and oncogenic transformation of NIH 3T3 cells. Using microarray analysis, we have previously identified a role for platelet-derived growth factor receptor alpha (PDGFRalpha) in Galpha12-mediated cell growth (R. N. Kumar et al., Cell Biochem. Biophys. 41:63-73, 2004). In the present study, we report that Galpha12QL stimulates the functional expression of PDGFRalpha and demonstrate that the expression of PDGFRalpha by Galpha12QL is dependent on the small GTPase Rho. Our results indicate that it is cell type independent as the transient expression of Galpha12QL or the activation of Galpha12-coupled receptors stimulates the expression of PDGFRalpha in NIH 3T3 as well as in human astrocytoma 1321N1 cells. Furthermore, we demonstrate the presence of an autocrine loop involving PDGF-A and PDGFRalpha in Galpha12QL-transformed cells. Analysis of the functional consequences of the Galpha12-PDGFRalpha signaling axis indicates that Galpha12 stimulates the phosphatidylinositol 3-kinase (PI3K)-AKT signaling pathway through PDGFR. In addition, we show that Galpha12QL stimulates the phosphorylation of forkhead transcription factor FKHRL1 via AKT in a PDGFRalpha- and PI3K-dependent manner. Since AKT promotes cell growth by blocking the transcription of antiproliferative genes through the inhibitory phosphorylation of forkhead transcription factors, our results describe for the first time a PDGFRalpha-dependent signaling pathway involving PI3K-AKT-FKHRL1, regulated by Galpha12QL in promoting cell growth. Consistent with this view, we demonstrate that the expression of a dominant negative mutant of PDGFRalpha attenuated Galpha12-mediated neoplastic transformation of NIH 3T3 cells. PMID- 16354681 TI - Defects in yolk sac vasculogenesis, chorioallantoic fusion, and embryonic axis elongation in mice with targeted disruption of Yap65. AB - YAP is a multifunctional adapter protein and transcriptional coactivator with several binding partners well described in vitro and in cell culture. To explore in vivo requirements for YAP, we generated mice carrying a targeted disruption of the Yap gene. Homozygosity for the Yap(tm1Smil) allele (Yap-/-) caused developmental arrest around E8.5. Phenotypic characterization revealed a requirement for YAP in yolk sac vasculogenesis. Yolk sac endothelial and erythrocyte precursors were specified as shown by histology, PECAM1 immunostaining, and alpha globin expression. Nonetheless, development of an organized yolk sac vascular plexus failed in Yap-/- embryos. In striking contrast, vasculogenesis proceeded in both the allantois and the embryo proper. Mutant embryos showed patterned gene expression domains along the anteroposterior neuraxis, midline, and streak/tailbud. Despite this evidence of proper patterning and tissue specification, Yap-/- embryos showed developmental perturbations that included a notably shortened body axis, convoluted anterior neuroepithelium, caudal dysgenesis, and failure of chorioallantoic fusion. These results reveal a vital requirement for YAP in the developmental processes of yolk sac vasculogenesis, chorioallantoic attachment, and embryonic axis elongation. PMID- 16354680 TI - Nutrients suppress phosphatidylinositol 3-kinase/Akt signaling via raptor dependent mTOR-mediated insulin receptor substrate 1 phosphorylation. AB - Nutritional excess and/or obesity represent well-known predisposition factors for the development of non-insulin-dependent diabetes mellitus (NIDDM). However, molecular links between obesity and NIDDM are only beginning to emerge. Here, we demonstrate that nutrients suppress phosphatidylinositol 3 (PI3)-kinase/Akt signaling via Raptor-dependent mTOR (mammalian target of rapamycin)-mediated phosphorylation of insulin receptor substrate 1 (IRS-1). Raptor directly binds to and serves as a scaffold for mTOR-mediated phosphorylation of IRS-1 on Ser636/639. These serines lie close to the Y(632)MPM motif that is implicated in the binding of p85alpha/p110alpha PI3-kinase to IRS-1 upon insulin stimulation. Phosphomimicking mutations of these serines block insulin-stimulated activation of IRS-1-associated PI3-kinase. Knockdown of Raptor as well as activators of the LKB1/AMPK pathway, such as the widely used antidiabetic compound metformin, suppress IRS-1 Ser636/639 phosphorylation and reverse mTOR-mediated inhibition on PI3-kinase/Akt signaling. Thus, diabetes-related hyperglycemia hyperactivates the mTOR pathway and may lead to insulin resistance due to suppression of IRS-1 dependent PI3-kinase/Akt signaling. PMID- 16354682 TI - Distinct roles of phosphoinositide-3 kinase and phospholipase Cgamma2 in B-cell receptor-mediated signal transduction. AB - During B-cell receptor (BCR) signaling, phosphoinositide-3 kinase (PI3K) is thought to function upstream of phospholipase Cgamma2 (PLCgamma2). PLCgamma2 deficiency specifically impedes transitional type 2 (T2) to follicular (FO) mature B-cell transition. Here, we demonstrate that PI3K deficiency specifically impaired T2-to-FO mature B-cell transition and marginal zone B-cell development. Furthermore, we investigated the functional relationship between PI3K and PLCgamma2 using PI3K-/-, PLCgamma2-/-, and PI3K-/- PLCgamma2-/- B cells. Interestingly, PLCgamma2 deficiency had no effect on BCR-mediated PI3K activation, whereas PI3K deficiency only partially blocked activation of PLCgamma2. Moreover, whereas PI3K-/- PLCgamma2-/- double deficiency did not affect hematopoiesis, it resulted in embryonic lethality. PI3K-/- PLCgamma2-/- fetal liver cells transplanted into B-cell null JAK3-/- mice failed to restore development of peripheral B cells and failed to progress through early B-cell development at the pro-B- to pre-B-cell transition, a more severe phenotype than was observed with either PI3K or PLCgamma2 single-deficiency B cells. Consistent with this finding, BCR signaling was more severely impaired in the absence of both PI3K and PLCgamma2 genes than in the absence of either one alone. Taken together, these results demonstrate that whereas PI3K functions upstream of PLCgamma2, activation of PLCgamma2 can occur independently of PI3K and that PI3K and PLCgamma2 also have distinct functions in BCR signal transduction. PMID- 16354683 TI - Distinct utilization of effectors and biological outcomes resulting from site specific Ras activation: Ras functions in lipid rafts and Golgi complex are dispensable for proliferation and transformation. AB - Ras proteins are distributed in different types of plasma membrane microdomains and endomembranes. However, how microlocalization affects the signals generated by Ras and its subsequent biological outputs is largely unknown. We have approached this question by selectively targeting RasV12 to different cellular sublocalizations. We show here that compartmentalization dictates Ras utilization of effectors and the intensity of its signals. Activated Ras can evoke enhanced proliferation and transformation from most of its platforms, with the exception of the Golgi complex. Furthermore, signals that promote survival emanate primarily from the endoplasmic reticulum pool. In addition, we have investigated the need for the different pools of endogenous Ras in the conveyance of upstream mitogenic and transforming signals. Using targeted RasN17 inhibitory mutants and in physiological contexts such as H-Ras/N-Ras double knockout fibroblasts, we demonstrate that Ras functions at lipid rafts and at the Golgi complex are fully dispensable for proliferation and transformation. PMID- 16354684 TI - PTF1 is an organ-specific and Notch-independent basic helix-loop-helix complex containing the mammalian Suppressor of Hairless (RBP-J) or its paralogue, RBP-L. AB - PTF1 is a trimeric transcription factor essential to the development of the pancreas and to the maintenance of the differentiated state of the adult exocrine pancreas. It comprises a dimer of P48/PTF1a (a pancreas and neural restricted basic helix-loop-helix [bHLH] protein) and a class A bHLH protein, together with a third protein that we show can be either the mammalian Suppressor of Hairless (RBP-J) or its paralogue, RBP-L. In mature acinar cells, PTF1 exclusively contains the RBP-L isoform and is bound to the promoters of acinar specific genes. P48 interacts with the RBP subunit primarily through two short conserved tryptophan-containing motifs, similar to the motif of the Notch intracellular domain (NotchIC) that interacts with RBP-J. The transcriptional activities of the J and L forms of PTF1 are independent of Notch signaling, because P48 occupies the NotchIC docking site on RBP-J and RBP-L does not bind the NotchIC. Mutations that delete one or both of the RBP-interacting motifs of P48 eliminate RBP binding and are associated with a human genetic disorder characterized by pancreatic and cerebellar agenesis, which indicates that the association of P48 and RBPs is required for proper embryonic development. The presence of related peptide motifs in other transcription factors indicates a broader Notch independent function for RBPJ/SU(H). PMID- 16354685 TI - Alternative pathways for the repair of RAG-induced DNA breaks. AB - RAG1 and RAG2 cleave DNA to generate blunt signal ends and hairpin coding ends at antigen receptor loci in lymphoid cells. During V(D)J recombination, repair of these RAG-generated double-strand breaks (DSBs) by the nonhomologous end-joining (NHEJ) pathway contributes substantially to the antigen receptor diversity necessary for immune system function, although recent evidence also supports the ability of RAG-generated breaks to undergo homology-directed repair (HDR). We have determined that RAG-generated chromosomal breaks can be repaired by pathways other than NHEJ in mouse embryonic stem (ES) cells, although repair by these pathways occurs at a significantly lower frequency than NHEJ. HDR frequency was estimated to be >or=40-fold lower than NHEJ frequency for both coding end and signal end reporters. Repair by single-strand annealing was estimated to occur at a comparable or lower frequency than HDR. As expected, V(D)J recombination was substantially impaired in cells deficient for the NHEJ components Ku70, XRCC4, and DNA-PKcs. Concomitant with decreased NHEJ, RAG-induced HDR was increased in each of the mutants, including cells lacking DNA-PKcs, which has been implicated in hairpin opening. HDR was increased to the largest extent in Ku70-/- cells, implicating the Ku70/80 DNA end-binding protein in regulating pathway choice. Thus, RAG-generated DSBs are typically repaired by the NHEJ pathway in ES cells, but in the absence of NHEJ components, a substantial fraction of breaks can be efficiently channeled into alternative pathways in these cells. PMID- 16354686 TI - Nox2 and Rac1 regulate H2O2-dependent recruitment of TRAF6 to endosomal interleukin-1 receptor complexes. AB - Reactive oxygen species (ROS) generated by NADPH oxidases (Nox) have been implicated in the regulation of signal transduction. However, the cellular mechanisms that link Nox activation with plasma membrane receptor signaling remain poorly defined. We have found that Nox2-derived ROS influence the formation of an active interleukin-1 (IL-1) receptor complex in the endosomal compartment by directing the H2O2-dependent binding of TRAF6 to the IL-1R1/MyD88 complex. Clearance of both superoxide and H2O2 from within the endosomal compartment significantly abrogated IL-1beta-dependent IKK and NF-kappaB activation. MyD88-dependent endocytosis of IL-1R1 following IL-1beta binding was required for the redox-dependent formation of an active endosomal receptor complex competent for IKK and NF-kappaB activation. Small interfering RNAs to either MyD88 or Rac1 inhibited IL-1beta induction of endosomal superoxide and NF kappaB activation. However, MyD88 and Rac1 appear to be recruited independently to IL-1R1 following ligand stimulation. In this context, MyD88 binding was required for inducing endocytosis of IL-1R1 following ligand binding, while Rac1 facilitated the recruitment of Nox2 into the endosomal compartment and subsequent redox-dependent recruitment of TRAF6 to the MyD88/IL-1R1 complex. The identification of Nox-active endosomes helps explain how subcellular compartmentalization of redox signals can be used to direct receptor activation from the plasma membrane. PMID- 16354687 TI - The forkhead transcription factor FoxI1 remains bound to condensed mitotic chromosomes and stably remodels chromatin structure. AB - All forkhead (Fox) proteins contain a highly conserved DNA binding domain whose structure is remarkably similar to the winged-helix structures of histones H1 and H5. Little is known about Fox protein binding in the context of higher-order chromatin structure in living cells. We created a stable cell line expressing FoxI1-green fluorescent protein (GFP) or FoxI1-V5 fusion proteins under control of the reverse tetracycline-controlled transactivator doxycycline inducible system and found that unlike most transcription factors, FoxI1 remains bound to the condensed chromosomes during mitosis. To isolate DNA fragments directly bound by the FoxI1 protein within living cells, we performed chromatin immunoprecipitation assays (ChIPs) with antibodies to either enhanced GFP or the V5 epitope and subcloned the FoxI1-enriched DNA fragments. Sequence analyses indicated that 88% (106/121) of ChIP sequences contain the consensus binding sites for all Fox proteins. Testing ChIP sequences with a quantitative DNase I hypersensitivity assay showed that FoxI1 created stable DNase I sensitivity changes in condensed chromosomes. The majority of ChIP targets and random targets increased in resistance to DNase I in FoxI1-expressing cells, but a small number of targets became more accessible to DNase I. Consistently, the accessibility of micrococcal nuclease to chromatin was generally inhibited. Micrococcal nuclease partial digestion generated a ladder in which all oligonucleosomes were slightly longer than those observed with the controls. On the basis of these findings, we propose that FoxI1 is capable of remodeling chromatin higher-order structure and can stably create site-specific changes in chromatin to either stably create or remove DNase I hypersensitive sites. PMID- 16354688 TI - A family of human zinc finger proteins that bind methylated DNA and repress transcription. AB - In vertebrates, densely methylated DNA is associated with inactive transcription. Actors in this process include proteins of the MBD family that can recognize methylated CpGs and repress transcription. Kaiso, a structurally unrelated protein, has also been shown to bind methylated CGCGs through its three Kruppel like C2H2 zinc fingers. The human genome contains two uncharacterized proteins, ZBTB4 and ZBTB38, that contain Kaiso-like zinc fingers. We report that ZBTB4 and ZBTB38 bind methylated DNA in vitro and in vivo. Unlike Kaiso, they can bind single methylated CpGs. When transfected in mouse cells, the proteins colocalize with foci of heavily methylated satellite DNA and become delocalized upon loss of DNA methylation. Chromatin immunoprecipitation suggests that both of these proteins specifically bind to the methylated allele of the H19/Igf2 differentially methylated region. ZBTB4 and ZBTB38 repress the transcription of methylated templates in transfection assays. The two genes have distinct tissue specific expression patterns, but both are highly expressed in the brain. Our results reveal the existence of a family of Kaiso-like proteins that bind methylated CpGs. Like proteins of the MBD family, they are able to repress transcription in a methyl-dependent manner, yet their tissue-specific expression pattern suggests nonoverlapping functions. PMID- 16354689 TI - Mice with a targeted disruption of the Cl-/HCO3- exchanger AE3 display a reduced seizure threshold. AB - Neuronal activity results in significant pH shifts in neurons, glia, and interstitial space. Several transport mechanisms are involved in the fine-tuning and regulation of extra- and intracellular pH. The sodium-independent electroneutral anion exchangers (AEs) exchange intracellular bicarbonate for extracellular chloride and thereby lower the intracellular pH. Recently, a significant association was found with the variant Ala867Asp of the anion exchanger AE3, which is predominantly expressed in brain and heart, in a large cohort of patients with idiopathic generalized epilepsy. To analyze a possible involvement of AE3 dysfunction in the pathogenesis of seizures, we generated an AE3-knockout mouse model by targeted disruption of Slc4a3. AE3-knockout mice were apparently healthy, and neither displayed gross histological and behavioral abnormalities nor spontaneous seizures or spike wave complexes in electrocorticograms. However, the seizure threshold of AE3-knockout mice exposed to bicuculline, pentylenetetrazole, or pilocarpine was reduced, and seizure induced mortality was significantly increased compared to wild-type littermates. In the pyramidal cell layer of the hippocampal CA3 region, where AE3 is strongly expressed, disruption of AE3 abolished sodium-independent chloride-bicarbonate exchange. These findings strongly support the hypothesis that AE3 modulates seizure susceptibility and, therefore, are of significance for understanding the role of intracellular pH in epilepsy. PMID- 16354690 TI - Tissue-specific differences of p53 inhibition by Mdm2 and Mdm4. AB - The function of the p53 tumor suppressor to inhibit proliferation or initiate apoptosis is often abrogated in tumor cells. Mdm2 and its homolog, Mdm4, are critical inhibitors of p53 that are often overexpressed in human tumors. In mice, loss of Mdm2 or Mdm4 leads to embryonic lethal phenotypes that are completely rescued by concomitant loss of p53. To examine the role of Mdm2 and Mdm4 in a temporal and tissue-specific manner and to determine the relationships of these inhibitors to each other, we generated conditional alleles. We deleted Mdm2 and Mdm4 in cardiomyocytes, since proliferation and apoptosis are important processes in heart development. Mice lacking Mdm2 in the heart were embryonic lethal and showed defects at the time recombination occurred. A critical number of cardiomyocytes were lost by embryonic day 13.5, resulting in heart failure. This phenotype was completely rescued by deletion of p53. Mice lacking Mdm4 in the heart were born at the correct ratio and appeared to be normal. Our studies provide the first direct evidence that Mdm2 can function in the absence of Mdm4 to regulate p53 activity in a tissue-specific manner. Moreover, Mdm4 cannot compensate for the loss of Mdm2 in heart development. PMID- 16354691 TI - Kaiso-deficient mice show resistance to intestinal cancer. AB - Kaiso is a BTB domain protein that associates with the signaling molecule p120 catenin and binds to the methylated sequence mCGmCG or the nonmethylated sequence CTGCNA to modulate transcription. In Xenopus laevis, xKaiso deficiency leads to embryonic death accompanied by premature gene activation in blastulae and upregulation of the xWnt11 gene. Kaiso has also been proposed to play an essential role in mammalian synapse-specific transcription. We disrupted the Kaiso gene in mice to assess its role in mammalian development. Kaiso-null mice were viable and fertile, with no detectable abnormalities of development or gene expression. However, when crossed with tumor-susceptible Apc(Min/+) mice, Kaiso null mice showed a delayed onset of intestinal tumorigenesis. Kaiso was found to be upregulated in murine intestinal tumors and is expressed in human colon cancers. Our data suggest that Kaiso plays a role in intestinal cancer and may therefore represent a potential target for therapeutic intervention. PMID- 16354692 TI - Notch activation is an early and critical event during T-Cell leukemogenesis in Ikaros-deficient mice. AB - The Ikaros transcription factor is both a key regulator of lymphocyte differentiation and a tumor suppressor in T lymphocytes. Mice carrying a hypomorphic mutation (Ik(L/L)) in the Ikaros gene all develop thymic lymphomas. Ik(L/L) tumors always exhibit strong activation of the Notch pathway, which is required for tumor cell proliferation in vitro. Notch activation occurs early in tumorigenesis and may precede transformation, as ectopic expression of the Notch targets Hes-1 and Deltex-1 is detected in thymocytes from young Ik(L/L) mice with no overt signs of transformation. Notch activation is further amplified by secondary mutations that lead to C-terminal truncations of Notch 1. Strikingly, restoration of Ikaros activity in tumor cells leads to a rapid and specific downregulation of Notch target gene expression and proliferation arrest. Furthermore, Ikaros binds to the Notch-responsive element in the Hes-1 promoter and represses Notch-dependent transcription from this promoter. Thus, Ikaros mediated repression of Notch target gene expression may play a critical role in defining the tumor suppressor function of this factor. PMID- 16354693 TI - Oxidative and electrophilic stresses activate Nrf2 through inhibition of ubiquitination activity of Keap1. AB - The Keap1-Nrf2 system is the major regulatory pathway of cytoprotective gene expression against oxidative and/or electrophilic stresses. Keap1 acts as a stress sensor protein in this system. While Keap1 constitutively suppresses Nrf2 activity under unstressed conditions, oxidants or electrophiles provoke the repression of Keap1 activity, inducing the Nrf2 activation. However, the precise molecular mechanisms behind the liberation of Nrf2 from Keap1 repression in the presence of stress remain to be elucidated. We hypothesized that oxidative and electrophilic stresses induce the nuclear accumulation of Nrf2 by affecting the Keap1-mediated rapid turnover of Nrf2, since such accumulation was diminished by the protein synthesis inhibitor cycloheximide. While both the Cys273 and Cys288 residues of Keap1 are required for suppressing Nrf2 nuclear accumulation, treatment of cells with electrophiles or mutation of these cysteine residues to alanine did not affect the association of Keap1 with Nrf2 either in vivo or in vitro. Rather, these treatments impaired the Keap1-mediated proteasomal degradation of Nrf2. These results support the contention that Nrf2 protein synthesized de novo after exposure to stress accumulates in the nucleus by bypassing the Keap1 gate and that the sensory mechanism of oxidative and electrophilic stresses is closely linked to the degradation mechanism of Nrf2. PMID- 16354694 TI - Mitogen-activated protein kinase cascade-mediated histone H3 phosphorylation is critical for telomerase reverse transcriptase expression/telomerase activation induced by proliferation. AB - Telomerase activity and telomerase reverse transcriptase (hTERT), the key component of the telomerase complex, are tightly proliferation regulated in normal and malignant cells both in vitro and in vivo; however, underlying mechanisms are unclear. In the present study, we identified mitogen-activated protein kinase (MAPK) cascade-mediated histone H3 ser10 phosphorylation to be a molecular link between proliferation and induction of hTERT/telomerase activity. In normal human T lymphocytes and fibroblasts, growth or stress stimuli known to drive H3 phosphorylation through the MAPK signaling induce hTERT expression and/or telomerase activity that was preceded by phosphorylated histone H3 (ser10) at the hTERT promoter. Blockade of the MAPK-triggered H3 phosphorylation significantly abrogates hTERT induction and ser10 phosphorylation at this promoter. However, H3 ser10 phosphorylation alone resulted in low, transient hTERT induction, as seen in fibroblasts, whereas H3 phosphorylation followed by its acetylation at lys14 robustly trans-activated the hTERT gene accompanying constitutive telomerase activity in normal and malignant T cells. H3 acetylation without phosphorylation similarly exerted weak effects on hTERT expression. These results define H3 phosphorylation as a key to hTERT transactivation induced by proliferation and reveal a fundamental mechanism for telomerase regulation in both normal human cells and transformed T cells. PMID- 16354695 TI - Procollagen C proteinase enhancer 1 genes are important determinants of the mechanical properties and geometry of bone and the ultrastructure of connective tissues. AB - Procollagen C proteinases (pCPs) cleave type I to III procollagen C propeptides as a necessary step in assembling the major fibrous components of vertebrate extracellular matrix. The protein PCOLCE1 (procollagen C proteinase enhancer 1) is not a proteinase but can enhance the activity of pCPs approximately 10-fold in vitro and has reported roles in inhibiting other proteinases and in growth control. Here we have generated mice with null alleles of the PCOLCE1 gene, Pcolce, to ascertain in vivo roles. Although Pcolce-/- mice are viable and fertile, Pcolce-/- male, but not female, long bones are more massive and have altered geometries that increase resistance to loading, compared to wild type. Mechanical testing indicated inferior material properties of Pcolce-/- male long bone, apparently compensated for by the adaptive changes in bone geometry. Male and female Pcolce-/- vertebrae both appeared to compensate for inferior material properties with thickened and more numerous trabeculae and had a uniquely altered morphology in deposited mineral. Ultrastructurally, Pcolce-/- mice had profoundly abnormal collagen fibrils in both mineralized and nonmineralized tissues. In Pcolce-/- tendon, 100% of collagen fibrils had deranged morphologies, indicating marked functional effects in this tissue. Thus, PCOLCE1 is an important determinant of bone mechanical properties and geometry and of collagen fibril morphology in mammals, and the human PCOLCE1 gene is identified as a candidate for phenotypes with defects in such attributes in humans. PMID- 16354696 TI - Drosophila Paf1 modulates chromatin structure at actively transcribed genes. AB - The Paf1 complex in yeast has been reported to influence a multitude of steps in gene expression through interactions with RNA polymerase II (Pol II) and chromatin-modifying complexes; however, it is unclear which of these many activities are primary functions of Paf1 and are conserved in metazoans. We have identified and characterized the Drosophila homologs of three subunits of the yeast Paf1 complex and found striking differences between the yeast and Drosophila Paf1 complexes. We demonstrate that although Drosophila Paf1, Rtf1, and Cdc73 colocalize broadly with actively transcribing, phosphorylated Pol II, and all are recruited to activated heat shock genes with similar kinetics; Rtf1 does not appear to be a stable part of the Drosophila Paf1 complex. RNA interference (RNAi)-mediated depletion of Paf1 or Rtf1 leads to defects in induction of Hsp70 RNA, but tandem RNAi-chromatin immunoprecipitation assays show that loss of neither Paf1 nor Rtf1 alters the density or distribution of phosphorylated Pol II on the active Hsp70 gene. However, depletion of Paf1 reduces trimethylation of histone H3 at lysine 4 in the Hsp70 promoter region and significantly decreases the recruitment of chromatin-associated factors Spt6 and FACT, suggesting that Paf1 may manifest its effects on transcription through modulating chromatin structure. PMID- 16354697 TI - Focal adhesion kinase is a substrate and downstream effector of SHP-2 complexed with Helicobacter pylori CagA. AB - Infection with cagA-positive Helicobacter pylori (H. pylori) is associated with atrophic gastritis, peptic ulcer, and gastric adenocarcinoma. The cagA gene product CagA is translocated from H. pylori into gastric epithelial cells and undergoes tyrosine phosphorylation by Src family kinases (SFKs). Tyrosine phosphorylated CagA binds and activates SHP-2 phosphatase and the C-terminal Src kinase (Csk) while inducing an elongated cell shape termed the "hummingbird phenotype." Here we show that CagA reduces the level of focal adhesion kinase (FAK) tyrosine phosphorylation in gastric epithelial cells. The decrease in phosphorylated FAK is due to SHP-2-mediated dephosphorylation of FAK at the activating phosphorylation sites, not due to Csk-dependent inhibition of SFKs, which phosphorylate FAK. Coexpression of constitutively active FAK with CagA inhibits induction of the hummingbird phenotype, whereas expression of dominant negative FAK elicits an elongated cell shape characteristic of the hummingbird phenotype. These results indicate that inhibition of FAK by SHP-2 plays a crucial role in the morphogenetic activity of CagA. Impaired cell adhesion and increased motility by CagA may be involved in the development of gastric lesions associated with cagA-positive H. pylori infection. PMID- 16354698 TI - Akt-mediated YB-1 phosphorylation activates translation of silent mRNA species. AB - YB-1 is a broad-specificity RNA-binding protein that is involved in regulation of mRNA transcription, splicing, translation, and stability. In both germinal and somatic cells, YB-1 and related proteins are major components of translationally inactive messenger ribonucleoprotein particles (mRNPs) and are mainly responsible for storage of mRNAs in a silent state. However, mechanisms regulating the repressor activity of YB-1 are not well understood. Here we demonstrate that association of YB-1 with the capped 5' terminus of the mRNA is regulated via phosphorylation by the serine/threonine protein kinase Akt. In contrast to its nonphosphorylated form, phosphorylated YB-1 fails to inhibit cap-dependent but not internal ribosome entry site-dependent translation of a reporter mRNA in vitro. We also show that similar to YB-1, Akt is associated with inactive mRNPs and that activated Akt may relieve translational repression of the YB-1-bound mRNAs. Using Affymetrix microarrays, we found that many of the YB-1-associated messages encode stress- and growth-related proteins, raising the intriguing possibility that Akt-mediated YB-1 phosphorylation could, in part, increase production of proteins regulating cell proliferation, oncogenic transformation, and stress response. PMID- 16354699 TI - Innate immune responses in NF-kappaB-repressing factor-deficient mice. AB - NF-kappaB-repressing factor (NRF) is a transcriptional silencer protein that specifically counteracts the basal activity of several NF-kappaB-dependent promoters by direct binding to specific neighboring DNA sequences. In cell culture experiments, the reduction of NRF mRNA leads to a derepression of beta interferon, interleukin-8, and inducible nitric oxide synthase transcription. The X chromosome-located single-copy NRF gene is ubiquitously expressed and encodes a protein of 690 amino acids. The N-terminal part contains a nuclear localization signal, the DNA-binding domain, and the NF-kappaB-repressing domain, while the C terminal part is responsible for double-stranded RNA binding and nucleolar localization. To study the function of NRF in a systemic context, transgenic mice lacking the NRF gene were created. Against predictions from in vitro experiments, mice with a deletion of the NRF gene are viable and have a phenotype that is indistinguishable from wild-type mice, even after challenge with different pathogens. The data hint towards an unexpected functional redundancy of NRF. PMID- 16354700 TI - Direct interaction with Rab11a targets the epithelial Ca2+ channels TRPV5 and TRPV6 to the plasma membrane. AB - TRPV5 and TRPV6 are the most Ca2+-selective members of the transient receptor potential (TRP) family of cation channels and play a pivotal role in the maintenance of Ca2+ balance in the body. However, little is known about the mechanisms controlling the plasma membrane abundance of these channels to regulate epithelial Ca2+ transport. In this study, we demonstrated the direct and specific interaction of GDP-bound Rab11a with TRPV5 and TRPV6. Rab11a colocalized with TRPV5 and TRPV6 in vesicular structures underlying the apical plasma membrane of Ca2+-transporting epithelial cells. This GTPase recognized a conserved stretch in the carboxyl terminus of TRPV5 that is essential for channel trafficking. Furthermore, coexpression of GDP-locked Rab11a with TRPV5 or TRPV6 resulted in significantly decreased Ca2+ uptake, caused by diminished channel cell surface expression. Together, our data demonstrated the important role of Rab11a in the trafficking of TRPV5 and TRPV6. Rab11a exerts this function in a novel fashion, since it operates via direct cargo interaction while in the GDP bound configuration. PMID- 16354701 TI - Identifying optimal lipid raft characteristics required to promote nanoscale protein-protein interactions on the plasma membrane. AB - The dynamic lateral segregation of signaling proteins into microdomains is proposed to facilitate signal transduction, but the constraints on microdomain size, mobility, and diffusion that might realize this function are undefined. Here we interrogate a stochastic spatial model of the plasma membrane to determine how microdomains affect protein dynamics. Taking lipid rafts as representative microdomains, we show that reduced protein mobility in rafts segregates dynamically partitioning proteins, but the equilibrium concentration is largely independent of raft size and mobility. Rafts weakly impede small-scale protein diffusion but more strongly impede long-range protein mobility. The long range mobility of raft-partitioning and raft-excluded proteins, however, is reduced to a similar extent. Dynamic partitioning into rafts increases specific interprotein collision rates, but to maximize this critical, biologically relevant function, rafts must be small (diameter, 6 to 14 nm) and mobile. Intermolecular collisions can also be favored by the selective capture and exclusion of proteins by rafts, although this mechanism is generally less efficient than simple dynamic partitioning. Generalizing these results, we conclude that microdomains can readily operate as protein concentrators or isolators but there appear to be significant constraints on size and mobility if microdomains are also required to function as reaction chambers that facilitate nanoscale protein-protein interactions. These results may have significant implications for the many signaling cascades that are scaffolded or assembled in plasma membrane microdomains. PMID- 16354702 TI - Exploring the cellular activity of camptothecin-triple-helix-forming oligonucleotide conjugates. AB - Topoisomerase I is a ubiquitous DNA-cleaving enzyme and an important therapeutic target in cancer chemotherapy for camptothecins (CPTs). These drugs stimulate DNA cleavage by topoisomerase I but exhibit little sequence preference, inducing toxicity and side effects. A convenient strategy to confer sequence specificity consists of the linkage of topoisomerase poisons to DNA sequence recognition elements. In this context, triple-helix-forming oligonucleotides (TFOs) covalently linked to CPTs were investigated for the capacity to direct topoisomerase I-mediated DNA cleavage in cells. In the first part of our study, we showed that these optimized conjugates were able to regulate gene expression in cells upon the use of a Photinus pyralis luciferase reporter gene system. Furthermore, the formation of covalent topoisomerase I/DNA complexes by the TFO CPT conjugates was detected in cell nuclei. In the second part, we elucidated the molecular specificity of topoisomerase I cleavage by the conjugates by using modified DNA targets and in vitro cleavage assays. Mutations either in the triplex site or in the DNA duplex receptor are not tolerated; such DNA modifications completely abolished conjugate-induced cleavage all along the DNA. These results indicate that these conjugates may be further developed to improve chemotherapeutic cancer treatments by targeting topoisomerase I-induced DNA cleavage to appropriately chosen genes. PMID- 16354703 TI - Central role for protein targeting to glycogen in the maintenance of cellular glycogen stores in 3T3-L1 adipocytes. AB - Overexpression of the protein phosphatase 1 (PP1) subunit protein targeting to glycogen (PTG) markedly enhances cellular glycogen levels. In order to disrupt the endogenous PTG-PP1 complex, small interfering RNA (siRNA) constructs against PTG were identified. Infection of 3T3-L1 adipocytes with PTG siRNA adenovirus decreased PTG mRNA and protein levels by >90%. In parallel, PTG reduction resulted in a >85% decrease in glycogen levels 4 days after infection, supporting a critical role for PTG in glycogen metabolism. Total PP1, glycogen synthase, and GLUT4 levels, as well as insulin-stimulated signaling cascades, were unaffected. However, PTG knockdown reduced glycogen-targeted PP1 protein levels, corresponding to decreased cellular glycogen synthase- and phosphorylase-directed PP1 activity. Interestingly, GLUT1 levels and acute insulin-stimulated glycogen synthesis rates were increased two- to threefold, and glycogen synthase activation in the presence of extracellular glucose was maintained. In contrast, glycogenolysis rates were markedly increased, suggesting that PTG primarily acts to suppress glycogen breakdown. Cumulatively, these data indicate that disruption of PTG expression resulted in the uncoupling of PP1 activity from glycogen metabolizing enzymes, the enhancement of glycogenolysis, and a dramatic decrease in cellular glycogen levels. Further, they suggest that reduction of glycogen stores induced cellular compensation by several mechanisms, but ultimately these changes could not overcome the loss of PTG expression. PMID- 16354704 TI - Rhp51-dependent recombination intermediates that do not generate checkpoint signal are accumulated in Schizosaccharomyces pombe rad60 and smc5/6 mutants after release from replication arrest. AB - The Schizosaccharomyces pombe rad60 gene is essential for cell growth and is involved in repairing DNA double-strand breaks. Rad60 physically interacts with and is functionally related to the structural maintenance of chromosomes 5 and 6 (SMC5/6) protein complex. In this study, we investigated the role of Rad60 in the recovery from the arrest of DNA replication induced by hydroxyurea (HU). rad60-1 mutant cells arrested mitosis normally when treated with HU. Significantly, Rad60 function is not required during HU arrest but is required on release. However, the mutant cells underwent aberrant mitosis accompanied by irregular segregation of chromosomes, and DNA replication was not completed, as revealed by pulsed field gel electrophoresis. The deletion of rhp51 suppressed the aberrant mitosis of rad60-1 cells and caused mitotic arrest. These results suggest that Rhp51 and Rad60 are required for the restoration of a stalled or collapsed replication fork after release from the arrest of DNA replication by HU. The rad60-1 mutant was proficient in Rhp51 focus formation after release from the HU-induced arrest of DNA replication or DNA-damaging treatment. Furthermore, the lethality of a rad60 1 rqh1Delta double mutant was suppressed by the deletion of rhp51 or rhp57. These results suggest that Rad60 is required for recombination repair at a step downstream of Rhp51. We propose that Rhp51-dependent DNA structures that cannot activate the mitotic checkpoints accumulate in rad60-1 cells. PMID- 16354705 TI - Deletion of the gene encoding proprotein convertase 5/6 causes early embryonic lethality in the mouse. AB - PC5 belongs to the proprotein convertase family and activates precursor proteins by cleavage at basic sites during their transit through the secretory pathway and/or at the cell surface. These precursors include prohormones, proreceptors, growth factors, adhesion molecules, and viral glycoproteins. The Pcsk5 gene encodes two alternatively spliced isoforms, the soluble PC5A and transmembrane PC5B. We have carefully analyzed the expression of PC5 in the mouse during development and in adulthood by in situ hybridization, as well as in mouse tissues and various cell lines by quantitative reverse transcription-PCR. The data show that adrenal cortex and intestine are the richest sources of PC5A and PC5B, respectively. To better define the specific physiological roles of PC5, we have generated a mouse Pcsk5(Delta4)-deficient allele missing exon 4 that encodes the catalytic Asp173. While Delta4/+ heterozygotes were healthy and fertile, genotyping of progeny obtained from Delta4/+ interbreeding indicated that Delta4/Delta4 embryos died between embryonic days 4.5 and 7.5. These data demonstrate that Pcsk5 is an essential gene. PMID- 16354706 TI - Regulation of CD44 alternative splicing by SRm160 and its potential role in tumor cell invasion. AB - The multiple isoforms of the transmembrane glycoprotein CD44 are produced by alternative RNA splicing. Expression of CD44 isoforms containing variable 5 exon (v5) correlates with enhanced malignancy and invasiveness of some tumors. Here we demonstrate that SRm160, a splicing coactivator, regulates CD44 alternative splicing in a Ras-dependent manner. Overexpression of SRm160 stimulates inclusion of CD44 v5 when Ras is activated. Conversely, small interfering RNA (siRNA) mediated silencing of SRm160 significantly reduces v5 inclusion. Immunoprecipitation shows association of SRm160 with Sam68, a protein that also stimulates v5 inclusion in a Ras-dependent manner, suggesting that these two proteins interact to regulate CD44 splicing. Importantly, siRNA-mediated depletion of CD44 v5 decreases tumor cell invasion. Reduction of SRm160 by siRNA transfection downregulates the endogenous levels of CD44 isoforms, including v5, and correlates with a decrease in tumor cell invasiveness. PMID- 16354707 TI - The short-lived Matalpha2 transcriptional repressor is protected from degradation in vivo by interactions with its corepressors Tup1 and Ssn6. AB - The Matalpha2 (alpha2) protein is a transcriptional repressor necessary for the proper expression of cell type-specific genes in Saccharomyces cerevisiae. Like many transcription factors, alpha2 is rapidly degraded in vivo by the ubiquitin proteasome pathway. At least two different ubiquitin-dependent pathways target alpha2 for destruction, one of which recognizes the well-characterized Deg1 degradation determinant near the N terminus of the protein. Here we report that the alpha2 corepressors Tup1 and Ssn6 modify the in vivo degradation rate of alpha2. Tup1 modulates the metabolic stability of alpha2 by directly binding to the Deg1-containing region of the protein. TUP1 overexpression specifically stabilizes Deg1-containing proteins but not other substrates of the same ubiquitination enzymes that recognize Deg1. Point mutations in both alpha2 and Tup1 that compromise the alpha2-Tup1 binding interaction disrupt the ability of Tup1 to stabilize Deg1 proteins. The physical association between Tup1 and alpha2 competes with the ubiquitination machinery for access to the Deg1 signal. Finally, we observe that overproduction of both Tup1 and Ssn6, but not either alone, strongly stabilizes the endogenous alpha2 protein. From these results, we propose that the fraction of alpha2 found in active regulatory complexes with Tup1 and Ssn6 is spared from rapid proteolytic destruction and is stabilized relative to the uncomplexed pool of the protein. PMID- 16354708 TI - Replication past a trans-4-hydroxynonenal minor-groove adduct by the sequential action of human DNA polymerases iota and kappa. AB - The X-ray crystal structure of human DNA polymerase iota (Poliota) has shown that it differs from all known Pols in its dependence upon Hoogsteen base pairing for synthesizing DNA. Hoogsteen base pairing provides an elegant mechanism for synthesizing DNA opposite minor-groove adducts that present a severe block to synthesis by replicative DNA polymerases. Germane to this problem, a variety of DNA adducts form at the N2 minor-groove position of guanine. Previously, we have shown that proficient and error-free replication through the gamma-HOPdG (gamma hydroxy-1,N2-propano-2'-deoxyguanosine) adduct, which is formed from the reaction of acrolein with the N2 of guanine, is mediated by the sequential action of human Poliota and Polkappa, in which Poliota incorporates the nucleotide opposite the lesion site and Polkappa carries out the subsequent extension reaction. To test the general applicability of these observations to other adducts formed at the N2 position of guanine, here we examine the proficiency of human Poliota and Polkappa to synthesize past stereoisomers of trans-4-hydroxy-2-nonenal deoxyguanosine (HNE-dG). Even though HNE- and acrolein-modified dGs share common structural features, due to their increased size and other structural differences, HNE adducts are potentially more blocking for replication than gamma HOPdG. We show here that the sequential action of Poliota and Polkappa promotes efficient and error-free synthesis through the HNE-dG adducts, in which Poliota incorporates the nucleotide opposite the lesion site and Polkappa performs the extension reaction. PMID- 16354709 TI - Man-made versus female-made environment--will the real capacitation please stand up? AB - Two recent reviews discuss the importance of the female tract in regulating the function of the spermatozoon during its eventful journey to the site of fertilization in this journal. However, our understanding of the nature of this journey, specifically in the human, is remarkably poor. What is also clear is the discourse between what is likely happening in vivo and the design of our experiments in vitro. Our conclusion is that, to date, we have been studying the spermatozoa in the wrong environment, in the wrong way and at the wrong time. PMID- 16354710 TI - Using dietetic assistants to improve the outcome of hip fracture: a randomised controlled trial of nutritional support in an acute trauma ward. AB - OBJECTIVE: To examine how improved attention to nutritional status and dietary intake, achieved through the employment of dietetic assistants (DAs), will affect postoperative clinical outcome among elderly women with hip fracture. DESIGN: Open prospective randomised controlled trial, comparing conventional nursing care with the additional nutritional support provided by DA. SETTING: Thirty-eight bedded acute trauma ward in a teaching hospital. PARTICIPANTS: All but 11 of 344 consecutive admissions with acute nonpathological hip fracture were approached. Three hundred and eighteen (93%) agreed to inclusion. Sixteen were ineligible as they were immediately transferred to another acute ward, were managed conservatively or died preoperatively. PRIMARY OUTCOME MEASURE: Postoperative mortality in the acute trauma unit. SECONDARY OUTCOME MEASURES: Postoperative mortality at 4 months after fracture, length of stay, energy intake and nutritional status. RESULTS: DA-supported participants were less likely to die in the acute ward (4.1 versus 10.1%, P = 0.048). This effect was still apparent at 4 month follow-up (13.1 versus 22.9%, P = 0.036). DA-supported subjects had significantly better mean daily energy intake (1,105 kcal versus 756 kcal/24 h, 95% CI 259-440 kcal/24 h, P<0.001), significantly smaller reduction in mid-arm circumference during their inpatient stay (0.39 cm, P = 0.002) and nonsignificantly favourable results for other anthropometric and laboratory measurements. CONCLUSION: Dietetic or nutrition assistants are being introduced in units across the UK. This, the largest ever study of nutritional support after hip fracture, shows that their employment significantly reduced patients' risk of dying in the acute trauma unit; an effect that persisted at 4 month follow-up. PMID- 16354711 TI - Impact of zinc supplementation in children with acute diarrhoea in Turkey. AB - OBJECTIVE: Zinc deficiency is prevalent in children in developing countries. Supplemental zinc provides therapeutic benefits in diarrhoea. Our aim was to evaluate the effect of daily zinc supplementation for 14 days on diarrhoea duration, severity, and morbidity in children. METHODS: In a randomised, open label non-placebo controlled trial, we assessed the efficacy of providing zinc sulfate to 6-60 month old children with acute diarrhoea for 2 weeks followed by 3 months of morbidity surveillance. Children were randomly assigned to zinc (n = 150) and control (n = 130) groups and received 15-30 mg elemental zinc daily. RESULTS: Supplemented children had significantly improved plasma zinc levels by day 14 of therapy. Zinc deficiency was observed in 2.6% of the treatment and 3.3% of the control group. The mean duration of diarrhoea after starting supplementation was 3.02+/-2 days in the zinc group and 3.67+/-3.2 days in the control group. There was no significant difference in diarrhoea duration by treatment group (p>0.05). The number of stools after starting supplementation was 5.8+/-3.7 and 5.1+/-3.9 on day 1, 2.9+/-1.6 and 3.0+/-2.2 on day 2, and 1.8+/-1.1 and 1.6+/-0.9 on day 3 in the zinc and control groups, respectively. There was no significant difference in diarrhoea severity by treatment group (p>0.05). No significant effect was found on the incidence and prevalence of diarrhoea in the zinc compared with the control group. CONCLUSION: Our data indicate that supplementing children with acute diarrhoea in Turkey with 3 RDA of elemental zinc for 14 days improved neither diarrhoea duration nor severity despite significant increments in plasma zinc. PMID- 16354712 TI - The TRENDS Project: development of a methodology to reliably monitor the obesity epidemic in childhood. AB - AIMS: The government has set a target to halt the rise in childhood obesity in those aged under 11 by 2010, but no system is in place to ascertain if this has been achieved. We aimed to develop a simple and reproducible methodology to monitor trends in childhood obesity. METHODS: A purposive sample of 10 primary schools and three secondary schools was selected. Children were measured with parental "opt out" consent in reception class, year 4, and year 8 (ages 5, 9, and 13 years, respectively). Measurements were compared with those obtained locally in 1996-2001. Calculations were then performed to ascertain the sample size required to confidently identify a halt in the rise in obesity using three growth measures. RESULTS: A total of 999 children were measured with ascertainment of 95% in primary and 85% in secondary schools. The proportion of overweight and obese children aged 9 and 13 years had increased since 1996-2001, although only 9 year olds showed a significant rise. A general trend of an increase in obesity was observed with increasing age. Calculations showed that 1900-2400 children per age group are needed to detect a halt in the rise in obesity based on mean body mass index (BMI) standard deviation scores (SDS) by 2010 with 90% power, whereas 4200-10 500 children are needed for other measures. CONCLUSION: We have developed a simple, cost effective methodology for accurately measuring the epidemic and recommend the use of mean BMI SDS for demonstrating if a halt has been achieved. PMID- 16354713 TI - Incorporating interference into linkage analysis for experimental crosses. AB - The phenomenon of interference in genetic recombination is well-known and studied in a wide variety of organisms. Multilocus linkage analysis, which makes use of recombination patterns among all genetic markers simultaneously, is routinely used with data on humans and experimental organisms to build genetic maps. It is also used to try to determine the genes involved in traits of interest, such as common diseases. Most linkage analyses performed today ignore the occurrence of genetical interference. We present an extension to the Lander-Green algorithm for experimental crosses (backcross and intercross) to incorporate crossover interference according to the chi2 model. Simulation results show the impact of using this model on the accuracy of estimated genetic maps. PMID- 16354714 TI - The endocytic pathway and formation of the Wingless morphogen gradient. AB - Controlling the spread of morphogens is crucial for pattern formation during development. In the Drosophila wing disc, Wingless secreted at the dorsal-ventral compartment boundary forms a concentration gradient in receiving tissue, where it activates short- and long-range target genes. The glypican Dally-like promotes Wingless spreading by unknown mechanisms, while Dynamin-dependent endocytosis is thought to restrict Wingless spread. We have utilized short-term expression of dominant negative Rab proteins to examine the polarity of endocytic trafficking of Wingless and its receptors and to determine the relative contributions of endocytosis, degradation and recycling to the establishment of the Wingless gradient. Our results show that Wingless is internalized via two spatially distinct routes: one on the apical, and one on the basal, side of the disc. Both restrict the spread of Wingless, with little contribution from subsequent degradation or recycling. As previously shown for Frizzled receptors, depleting Arrow does not prevent Wingless from entering endosomes. We find that both Frizzled and Arrow are internalized mainly from the apical membrane. Thus, the basal Wingless internalization route must be independent of these proteins. We find that Dally-like is not required for Wingless spread when endocytosis is blocked, and propose that Dally-like promotes the spread of Wingless by directing it to lateral membranes, where its endocytosis is less efficient. Thus, subcellular localization of Wingless along the apical-basal axis of receiving cells may be instrumental in shaping the Wingless gradient. PMID- 16354715 TI - Convergence of Wnt and FGF signals in the genesis of posterior neural plate through activation of the Sox2 enhancer N-1. AB - The expression of the transcription factor gene Sox2 precisely marks the neural plate in various vertebrate species. We previously showed that the Sox2 expression prevailing in the neural plate of chicken embryos is actually regulated by the coordination of five phylogenetically conserved enhancers having discrete regional coverage, among which the 420-bp long enhancer N-1, active in the node-proximal region, is probably involved directly in the genesis of the posterior neural plate. We investigated the signaling systems regulating this enhancer, first identifying the 56-bp N-1 core enhancer (N-1c), which in a trimeric form recapitulates the activity of the enhancer N-1. Mutational analysis identified five blocks, A to E, that regulate the enhancer N-1c. Functional analysis of these blocks indicated that Wnt and FGF signals synergistically activate the enhancer through Blocks A-B, bound by Lef1, and Block D, respectively. Fgf8b and Wnt8c expressed in the organizer-primitive streak region account for the activity in the embryo. Block E is essential for the repression of the enhancer N-1c activity in the mesendodermal precursors. The enhancer N-1c is not affected by BMP signals. Thus, Wnt and FGF signals converge to activate Sox2 expression through the enhancer N-1c, revealing the direct involvement of the Wnt signal in the initiation of neural plate development. PMID- 16354716 TI - The conserved oligomeric Golgi complex acts in organ morphogenesis via glycosylation of an ADAM protease in C. elegans. AB - In C. elegans, the gonad acquires two U-shaped arms through directed migration of gonadal distal tip cells (DTCs). A member of the ADAM (a disintegrin and metalloprotease) family, MIG-17, is secreted from muscle cells and localizes to the gonadal basement membrane where it functions in DTC migration. Mutations in cogc-3 and cogc-1 cause misdirected DTC migration similar to that seen in mig-17 mutants. Here, we report that COGC-3 and COGC-1 proteins are homologous to mammalian COG-3/Sec34 and COG-1/ldlBp, respectively, two of the eight components of the conserved oligomeric Golgi (COG) complex required for Golgi function. Knockdown of any of the other six components by RNA interference also produces DTC migration defects, suggesting that the eight components function in a common pathway. COGC-3 and COGC-1 are required for the glycosylation and gonadal localization of MIG-17, but not for secretion of MIG-17 from muscle cells. Furthermore, COGC-3 requires MIG-17 activity for its action in DTC migration. Our findings demonstrate that COG complex-dependent glycosylation of an ADAM protease plays a crucial role in determining organ shape. PMID- 16354717 TI - Use of time-lapse imaging and dominant negative receptors to dissect the steroid receptor control of neuronal remodeling in Drosophila. AB - During metamorphosis, the reorganization of the nervous system of Drosophila melanogaster proceeds in part through remodeling of larval neurons. In this study, we used in-vitro imaging techniques and immunocytochemistry to track the remodeling of the thoracic ventral neurosecretory cells. Axons of these neurons prune their larval arbors early in metamorphosis and a larger, more extensive adult arbor is established via branch outgrowth. Expression of EcR dominant negative constructs and an EcR inverted repeat construct resulted in pruning defects of larval axon arbors and a lack of filopodia during pruning, but showed variable effects on outgrowth depending on the construct expressed. Cells expressing either UAS-EcR-B1(W650A) or UAS-EcR-A(W650A) lacked filopodia during the outgrowth period and formed a poorly branched, larval-like arbor in the adult. Cells expressing UAS-EcR-B1(F645A), UAS-EcR-B2(W650A) or UAS-IR-EcR (core) showed moderate filopodial activity and normal, albeit reduced, adult-like branching during outgrowth. These results are consistent with the role of activation versus derepression via EcR for successive phases of neuronal remodeling and suggest that functional ecdysone receptor is necessary for some, but not all, remodeling events. PMID- 16354718 TI - Chromosomal clustering and GATA transcriptional regulation of intestine-expressed genes in C. elegans. AB - We used mRNA tagging to identify genes expressed in the intestine of C. elegans. Animals expressing an epitope-tagged protein that binds the poly-A tail of mRNAs (FLAG::PAB-1) from an intestine-specific promoter (ges-1) were used to immunoprecipitate FLAG::PAB-1/mRNA complexes from the intestine. A total of 1938 intestine-expressed genes (P<0.001) were identified using DNA microarrays. First, we compared the intestine-expressed genes with those expressed in the muscle and germline, and identified 510 genes enriched in all three tissues and 624 intestine-, 230 muscle- and 1135 germ line-enriched genes. Second, we showed that the 1938 intestine-expressed genes were physically clustered on the chromosomes, suggesting that the order of genes in the genome is influenced by the effect of chromatin domains on gene expression. Furthermore, the commonly expressed genes showed more chromosomal clustering than the tissue-enriched genes, suggesting that chromatin domains may influence housekeeping genes more than tissue-specific genes. Third, in order to gain further insight into the regulation of intestinal gene expression, we searched for regulatory motifs. This analysis found that the promoters of the intestine genes were enriched for the GATA transcription factor consensus binding sequence. We experimentally verified these results by showing that the GATA motif is required in cis and that GATA transcription factors are required in trans for expression of these intestinal genes. PMID- 16354719 TI - Nitric oxide production is higher in rat cardiac microvessel endothelial cells than ventricular cardiomyocytes in baseline and hypoxic conditions: a comparative study. AB - The relative importance of endothelium- and cardiomyocyte-derived nitric oxide (NO) is unknown, with a lack of direct studies on cardiac microvessel endothelial cells (CMEC) and cardiomyocytes regarding relative cellular NO production. AIMS: To assess and compare baseline and hypoxia-induced NO and ONOO- production in cardiomyocytes and CMEC. METHODS: Rat cardiomyocytes were isolated, and cultured rat CMEC were purchased commercially. Hypoxia (+/- NOS inhibitors) was induced by mineral oil layering or hypoxic culture. NO and ONOO- were detected by FACS analysis of DAF-2/DA and DHR123, respectively. Total eNOS was determined by Western blot analysis. RESULTS: 1) Baseline NO production in CMEC was sevenfold (cultured cells) and 26-fold (isolated cells) higher than in cardiomyocytes, 2) eNOS expression was 22-fold higher in CMEC, 3) hypoxia increased NO production in both cell types, albeit to a larger extent in CMEC, 4) in hypoxic cardiomyocytes, nonselective NOS and iNOS-specific inhibition attenuated NO production, whereas in CMEC, iNOS-specific inhibition was ineffective, and 5) baseline ONOO- production was 2.2 times greater in CMEC than in cardiomyocytes. CONCLUSION: Using a novel approach, this study demonstrated that CMEC produce more baseline NO than cardiomyocytes, and that hypoxia activates NOS to increase NO production in both cell types. Baseline eNOS content was higher in CMEC than in cardiomyocytes, suggesting that differences in baseline NO production were eNOS associated. PMID- 16354720 TI - Endocytic pathways and biological effects induced by UVB-dependent or ligand dependent activation of the keratinocyte growth factor receptor. AB - UVB exposure of epidermal cells is known to trigger early and late molecular pathways dependent on receptor tyrosine kinases and reactive oxygen species (ROS). We have recently reported that UVB irradiation induces tyrosine phosphorylation, kinase activation, and internalization of the receptor for the keratinocyte growth factor (KGFR), a paracrine mediator of epithelial growth, differentiation, and survival. Here we analyzed in more detail the UVB-induced endocytic pathway of KGFR and the role of KGFR activation and internalization in regulating UVB-promoted apoptosis and cell cycle arrest. Immunogold electron microscopy and confocal analysis revealed that the UVB-induced endocytosis of KGFR occurs through clathrin-coated pits and that the internalized receptors are sorted to the degradative route and reach the lysosomal compartment with a timing similar to that induced by their ligand KGF. Treatment with the anti-oxidant N acetylcysteine inhibited KGFR endocytosis, suggesting that the receptor internalization is mediated by the intracellular production of ROS. The ligand independent KGFR endocytic pathway induced by UVB requires receptor kinase activity and tyrosine phosphorylation and involves transient receptor ubiquitination. Inhibition of KGFR activity reduces both the KGF-mediated proliferative response and the UVB-promoted apoptotic cell death, indicating a different effect of ligand-induced and UVB-induced KGFR triggering. In addition, receptor internalization leads to protection from apoptosis caused by UVB exposure. Finally, we compared directly the behavior of KGFR with that of the epidermal growth factor receptor (EGFR) upon UVB exposure. Surprisingly, biochemical and immunofluorescence analysis showed that EGFR, differently from KGFR, does not undergo UVB-induced tyrosine phosphorylation and internalization. Taken together, our results suggest a differential role of KGFR and EGFR in the response of epidermal cells to UVB possibly because KGFR endocytosis could be crucial for attenuation of survival signals in the suprabasal layers of human skin. PMID- 16354721 TI - Transgenic plant production of Cyanovirin-N, an HIV microbicide. AB - Cyanovirin-N (CV-N) is a microbicide candidate that inactivates a wide range of HIV strains by binding to gp120. Production of CV-N, or any protein microbicide, needs to be at extremely high levels and low cost to have an impact on global health. Thus, it is unlikely that fermentor-based systems will be suitable, including recombinant E. coli, where CV-N aggregates and dimers have consistently been found. Transgenic plants may provide a suitable expression system for protein microbicides, as production can be easily and economically scaled up. Here, Nicotiana tabacum was transformed with a gene encoding CV-N to explore proof of concept for the production of CV-N in transgenic plants. Plant-derived rCV-N was recoverable at levels of 130 ng/mg of fresh leaf tissue, or at least 0.85% of total soluble plant protein. Western blot analysis demonstrated that virtually all of the rCV-N was expressed in the desired monomeric form. Functionality was demonstrated by specific binding to gp120 and protection of T cells from in vitro HIV infection. Hydroponic culturing of transgenic plants demonstrated CV-N rhizosecretion at levels of 0.64 mug/ml hydroponic media after 24 days. Therefore, we suggest that transgenic plants have the potential to provide strategies for large-scale protein microbicide production. PMID- 16354722 TI - Calpain inhibitor MDL28170 modulates Abeta formation by inhibiting the formation of intermediate Abeta46 and protecting Abeta from degradation. AB - The observations that three major cleavages within the transmembrane domain of APP, namely, the gamma-cleavage, -cleavage, and the newly identified zeta cleavage, are involved in the generation of secreted Abeta40 and Abeta42 prompted us to determine how the calpain inhibitor III MDL 28170 influences these three cleavages and Abeta formation. With the use of a cell culture system, our data demonstrate that 1) at either high concentrations, or at a low range of concentrations, at early time points, MDL 28170 inhibits the formation of secreted Abeta40 and Abeta42. However, this effect is due to inhibition of the intermediate Abeta46 generation by zeta-cleavage and not due to direct inhibition of the gamma-cleavage that produces Abeta40/42 from Abeta46; 2) at low range of concentrations and at late time points, MDL 28170 causes an increase in secreted Abeta40/42 that likely results from inhibition of degradation of both the initial substrate, CTFbeta, and the final product, Abeta40/42, of gamma-secretase. These data strongly suggest that formation of Abeta46 is a key step in the gamma secretase mediated generation of Abeta40/42 and provide a new target for the development of Abeta inhibitors. These data also suggest that calpain and related proteases, which are sensitive to MDL 28170, play an important role in the accumulation of secreted Abeta. PMID- 16354723 TI - Dietary approaches to positively influence fetal determinants of adult health. AB - An imbalance between production and scavenging of oxidants is a commonality found in factors that result in fetal determinants that negatively affect adult health. We reasoned that a dietary intervention that promotes oxidant scavenging through phase 2 protein induction would have positive effects on fetal programming of adult health. Previously, we demonstrated that a diet containing broccoli sprouts high in glucoraphanin (Grn), precursor of the phase 2 protein inducer sulforaphane, decreased oxidative stress and associated problems in male spontaneously hypertensive stroke-prone rats (SHRsp). We hypothesized that placing females on a Grn-containing (Grn+) diet would have similar positive effects and that the adult offspring of such females would have lower blood pressures and less tissue inflammation than offspring from mothers on control diet. We demonstrate that female SHRsp on a Grn+ diet had decreased oxidative stress and associated problems such as hypertension than females on control diet. The offspring of females on Grn+ diet also had lower blood pressures and less tissue inflammation in adulthood regardless of diet, with offspring placed on a Grn+ diet having the best health outcomes. We conclude that reducing oxidative stress in pregnant females has profound outcomes in the health of their adult offspring. PMID- 16354724 TI - Long-term effect of therapeutic laser photocoagulation on gene expression in the eye. AB - Microarray-based gene expression analysis demonstrated that laser photocoagulation (LPC) of mouse eyes had a long-term effect on the expression of genes functionally related to tissue repair, cell migration, proliferation, ion, protein and nucleic acid metabolism, cell signaling, and angiogenesis. Six structural genes, including five crystallins (Cryaa, Cryba1, Crybb2, Crygc, Crygs) and keratin 1-12 (Krt1-12), the anti-angiogenic factor thrombospondin 1 (Tsp1), the retina- and brain-specific putative transcription factor tubby-like protein 1 (Tulp1), and transketolase (Tkt), a key enzyme in the pentose-phosphate pathway, were all shown to be up-regulated by real-time PCR and/or Western blotting. Immunohistochemistry localized five of these proteins to the laser lesions and surrounding tissue within the retina and pigmented epithelium. This is the first study demonstrating long-term changes in the expression of these genes associated with LPC. Therefore, it suggests that modulated gene expression might contribute to the long-term inhibitory effect of LPC. In addition, these genes present novel targets for gene-based therapies aimed at treating microangiopathies, especially diabetic retinopathy, a disease currently only treatable with LPC. PMID- 16354725 TI - Topochemical studies on modified lignin distribution in the xylem of Poplar (Populus spp.) after wounding. AB - BACKGROUND AND AIMS: Information on the influence of wounding on lignin synthesis and distribution in differentiating xylem tissue is still scarce. The present paper provides information on cell modifications with regard to wall ultrastructure and lignin distribution on cellular and subcellular levels in poplar after wounding. METHODS: Xylem of Populus spp. close to a wound was collected and processed for light microscopy, transmission electron microscopy and cellular UV microspectrophotometry. Cell wall modification with respect to lignin distribution was examined at different stages of wound tissue development. Scanning UV microspectrophotometry and point measurements were used to determine the lignin distribution. KEY RESULTS: Xylem fibres within a transition zone between differentiated xylem laid down prior to wounding and the tissues formed after wounding developed distinctively thickened secondary cell walls. Those modified walls and cell corners showed, on average, a higher lignin content and an inhomogeneous lignin distribution within the individual wall layers. CONCLUSIONS: The work presented shows that wounding of the xylem may induce a modified wall architecture and lignin distribution in tissues differentiating at the time of wounding. An increasing lignin content and distinctively thickened walls can contribute to improved resistance as part of the compartmentalization process. PMID- 16354726 TI - Motor coding in floccular climbing fibers. AB - The climbing fibers (CFs) that project from the dorsal cap of the inferior olive (IO) to the flocculus of the cerebellar cortex have been reported to be purely sensory, encoding "retinal slip." However, a clear oculomotor projection from the nucleus prepositus hypoglossi (NPH) to the IO has been shown. We therefore studied the sensorimotor information that is present in the CF signal. We presented rabbits with visual motion noise stimuli to break up the tight relation between instantaneous retinal slip and eye movement. Strikingly, the information about the motor behavior in the CF signal more than doubled that of the sensory component and was time-locked more tightly. The contribution of oculomotor signals was independently confirmed by analysis of spontaneous eye movements in the absence of visual input. The motor component of the CF code is essential to distinguish unexpected slip from self-generated slip, which is a prerequisite for proper oculomotor learning. PMID- 16354727 TI - Enhancement of excitatory synaptic transmission in spiny neurons after transient forebrain ischemia. AB - Spiny neurons in the neostriatum are highly vulnerable to ischemia. Enhancement of excitatory synaptic transmissions has been implicated in ischemia-induced excitotoxic neuronal death. Here we report that evoked excitatory postsynaptic currents in spiny neurons were potentiated after transient forebrain ischemia. The ischemia-induced potentiation in synaptic efficacy was associated with an enhancement of presynaptic release as demonstrated by an increase in the frequency of miniature excitatory postsynaptic currents (mEPSCs) and a decrease in the paired-pulse ratio. The amplitude of inward currents evoked by exogenous application of glutamate did not show significant changes after ischemia, suggesting that postsynaptic mechanism is not involved. The ischemia-induced increase in mEPSCs frequency was not affected by blockade of voltage-gated calcium channels, but it was eliminated in the absence of extracellular calcium. Bath application of ATP P2X receptor antagonist pyridoxal-phosphate-6-azophenyl 2',4'-disulfonic acid (PPADS) significantly reduced mEPSC frequency in ischemic neurons but had no effects on the control ones. Furthermore, the inhibitory effect of PPADS on ischemic neurons was abolished in Ca2+-free external solution. These results indicate that excitatory synaptic transmissions in spiny neurons are potentiated after ischemia via presynaptic mechanisms. Activation of P2X receptors and the consequent Ca2+ influx might contribute to the ischemia-induced facilitation of glutamate release. PMID- 16354728 TI - Temporal correlations in stochastic models of double bursting during simulated locomotion. AB - The output of the spinal central pattern generator for locomotion falls into two broad categories: alternation between antagonistic muscles and double bursting within muscles acting on multiple joints. We first model an alternating half center and then present two different models of double bursting. The first double bursting model consists of a central clock with an explicit one-to-one mapping between interneuron activity and model output. The second double-bursting model consists of a half-center with an added feedback neuron. Models are built using rate-coded leaky integrator neurons with slow self-inhibition. Structure-function relationships are explored by the addition of noise. The interaction of noise with the dynamics of each network creates a unique pattern of correlation between phases of the simulated cycle. The effects of noise can be explained by perturbation of deterministic versions of the networks. Three basic results were obtained: slow self-inhibitory currents lead to correlations between parts of the step cycle that are separated in time and network relative; model outputs are most sensitive to perturbations presented just before competitive switches in network activity, and clock-like models possess substantial symmetries within the correlation structure of burst durations, whereas the correlation structure of feedback models are asymmetric. Our models suggest that variability in burst length durations can be analyzed to make inferences about the structure of the spinal networks for locomotion. In particular, correlation patterns within double bursting outputs may yield important clues regarding the interaction between more central, clock-like networks and feedback from more peripheral interneurons. PMID- 16354729 TI - Histamine excites neonatal rat sympathetic preganglionic neurons in vitro via activation of H1 receptors. AB - The role of histamine in regulating excitability of sympathetic preganglionic neurons (SPNs) and the expression of histamine receptor mRNA in SPNs was investigated using whole-cell patch-clamp electrophysiological recording techniques combined with single-cell reverse transcriptase polymerase chain reaction (RT-PCR) in transverse neonatal rat spinal cord slices. Bath application of histamine (100 microM) or the H1 receptor agonist histamine trifluoromethyl toluidide dimaleate (HTMT; 10 microM) induced membrane depolarization associated with a decrease in membrane conductance in the majority (70%) of SPNs tested, via activation of postsynaptic H1 receptors negatively coupled to one or more unidentified K+ conductances. Histamine and HTMT application also induced or increased the amplitude and/or frequency of membrane potential oscillations in electrotonically coupled SPNs. The H2 receptor agonist dimaprit (10 microM) or the H3 receptor agonist imetit (100 nM) were without significant effect on the membrane properties of SPNs. Histamine responses were sensitive to the H1 receptor antagonist triprolidine (10 microM) and the nonselective potassium channel blocker barium (1 mM) but were unaffected by the H2 receptor antagonist tiotidine (10 microM) and the H3 receptor antagonist, clobenpropit (5 microM). Single cell RT-PCR revealed mRNA expression for H1 receptors in 75% of SPNs tested, with no expression of mRNA for H2, H3, or H4 receptors. These data represent the first demonstration of H1 receptor expression in SPNs and suggest that histamine acts to regulate excitability of these neurons via a direct postsynaptic effect on H1 receptors. PMID- 16354730 TI - Synchronous activity in cat visual cortex encodes collinear and cocircular contours. AB - We explored how contour information in primary visual cortex might be embedded in the simultaneous activity of multiple cells recorded with a 100-electrode array. Synchronous activity in cat visual cortex was more selective and predictable in discriminating between drifting grating and concentric ring stimuli than changes in firing rate. Synchrony was found even between cells with wholly different orientation preferences when their receptive fields were circularly aligned, and membership in synchronous groups was orientation and curvature dependent. The existence of synchrony between cocircular cells reinforces its role as a general mechanism for contour integration and shape detection as predicted by association field concepts. Our data suggest that cortical synchrony results from common and synchronous input from earlier visual areas and that it could serve to shape extrastriate response selectivity. PMID- 16354731 TI - Enhanced neuronal damage after ischemic insults in mice lacking Kir6.2-containing ATP-sensitive K+ channels. AB - Adenosine triphosphate (ATP)-sensitive potassium (KATP) channels, incorporating Kir6.x and sulfonylurea receptor subunits, are weak inward rectifiers that are thought to play a role in neuronal protection from ischemic insults. However, the involvement of Kir6.2-containing KATP channel in hippocampus and neocortex has not been tested directly. To delineate the physiological roles of Kir6.2 channels in the CNS, we used knockout (KO) mice that do not express Kir6.2. Immunocytochemical staining demonstrated that Kir6.2 protein was expressed robustly in hippocampal neurons of the wild-type (WT) mice and absent in the KO. To examine neuronal sensitivity to metabolic stress in vitro, and to ischemia in vivo, we 1) exposed hippocampal slices to transient oxygen and glucose deprivation (OGD) and 2) produced focal cerebral ischemia by middle cerebral artery occlusion (MCAO). Both slice and whole animal studies showed that neurons from the KO mice were severely damaged after anoxia or ischemia, whereas few injured neurons were observed in the WT, suggesting that Kir6.2 channels are necessary to protect neurons from ischemic insults. Membrane potential recordings from the WT CA1 pyramidal neurons showed a biphasic response to OGD; a brief hyperpolarization was followed by a small depolarization during OGD, with complete recovery within 30 min after returning to normoxic conditions. By contrast, CA1 pyramidal neurons from the KO mice were irreversibly depolarized by OGD exposure, without any preceding hyperpolarization. These data suggest that expression of Kir6.2 channels prevents prolonged depolarization of neurons resulting from acute hypoxic or ischemic insults, and thus protects these central neurons from the injury. PMID- 16354732 TI - Visual cortical recovery from reverse occlusion depends on concordant binocular experience. AB - The effects of early monocular deprivation on visual acuity and visual cortical responses can be reversed quickly if vision is restored to the deprived eye and the other eye is deprived instead, a procedure known as reverse occlusion. However, recovery of vision through the originally deprived eye (ODE) is not stable. Following re-opening of the recently deprived (originally nondeprived) eye (ONDE), vision in the ODE typically deteriorates rapidly, possibly because of competitive interactions, whereas vision in the ONDE also remains compromised, resulting in bilateral amblyopia, the reasons for which are unknown. Here we monitor the physiological changes in the visual cortex during recovery from reverse occlusion in a longitudinal study, using optical imaging of intrinsic signals and single-cell recording in anesthetized cats. We show that a brief period of just 4 days of concordant binocular vision intercalated between the two periods of monocular experience allows close to equal responses to develop through both eyes, both in terms of cortical territory and orientation selectivity. In contrast, with no binocular vision or discordant binocular experience, cortical territory dominated by the ONDE is significantly reduced, and orientation tuning of cells dominated by the ODE is wider than that of cells dominated by the ONDE. These results support the notion that a brief period of binocular vision is sufficient to prevent bilateral acuity loss caused by reverse occlusion. PMID- 16354733 TI - Spike-timing precision underlies the coding efficiency of auditory receptor neurons. AB - Sensory systems must translate incoming signals quickly and reliably so that an animal can act successfully in its environment. Even at the level of receptor neurons, however, functional aspects of the sensory encoding process are not yet fully understood. Specifically, this concerns the question how stimulus features and neural response characteristics lead to an efficient transmission of sensory information. To address this issue, we have recorded and analyzed spike trains from grasshopper auditory receptors, while systematically varying the stimulus statistics. The stimulus variations profoundly influenced the efficiency of neural encoding. This influence was largely attributable to the presence of specific stimulus features that triggered remarkably precise spikes whose trial to-trial timing variability was as low as 0.15 ms--one order of magnitude shorter than typical stimulus time scales. Precise spikes decreased the noise entropy of the spike trains, thereby increasing the rate of information transmission. In contrast, the total spike train entropy, which quantifies the variety of different spike train patterns, hardly changed when stimulus conditions were altered, as long as the neural firing rate remained the same. This finding shows that stimulus distributions that were transmitted with high information rates did not invoke additional response patterns, but instead displayed exceptional temporal precision in their neural representation. The acoustic stimuli that led to the highest information rates and smallest spike-time jitter feature pronounced sound-pressure deflections lasting for 2-3 ms. These upstrokes are reminiscent of salient structures found in natural grasshopper communication signals, suggesting that precise spikes selectively encode particularly important aspects of the natural stimulus environment. PMID- 16354734 TI - Informed consent--a new approach to drug regulation? PMID- 16354735 TI - Increased libido associated with quetiapine. AB - We report sexual symptoms in a 44 year old man with schizophrenia who was receiving treatment with the atypical antipsychotic quetiapine. The symptoms consisted of an increased drive to masturbate and spontaneous ejaculations at night time. We discuss the potential neuro-chemical mechanisms for this and argue that quetiapine's unique pharmacological profile may have contributed to the development of his symptoms. PMID- 16354736 TI - Development and validation of the Essen Intracerebral Haemorrhage Score. AB - BACKGROUND: Spontaneous intracerebral haemorrhage (ICH) accounts for the highest in-hospital mortality of all stroke types. Nevertheless, outcome is favourable in about 30% of patients. Only one model for the prediction of favourable outcome has been validated so far. OBJECTIVE: To describe the development and validation of the Essen ICH score. METHODS: Inception cohorts were assessed on the National Institutes of Health stroke scale (NIH-SS) on admission and after follow up of 100 days. On the basis of previously validated clinical variables, a simple clinical score was developed to predict mortality and complete recovery (Barthel index after 100 days>or=95) in 340 patients with acute ICH. Subscores for age (<60=0; 60-69=1; 70-79=2; >or=80=3), NIH-SS level of consciousness (alert=0; drowsy=1; stuporose=2; comatose=3), and NIH-SS total score (0-5=0; 6-10=1; 11 15=2; 16-20=3; >20 or coma=4) were combined into a prognostic scale with <3 predicting complete recovery and >7 predicting death. The score was subsequently validated in an external cohort of 371 patients. RESULTS: The Essen ICH score showed a high prognostic accuracy for complete recovery and death in both the development and validation cohort. For prediction of complete recovery on the Barthel index after 100 days, the Essen ICH score was superior to the physicians' prognosis and to two previous prognostic scores developed for a slightly modified outcome. CONCLUSIONS: The Essen ICH score provides an easy to use scale for outcome prediction following ICH. Its high positive predictive values for adverse outcomes and easy applicability render it useful for individual prognostic indications or the design of clinical studies. In contrast, physicians tended to predict outcome too pessimistically. PMID- 16354737 TI - Prevalence and intensity of pain after stroke: a population based study focusing on patients' perspectives. AB - OBJECTIVE: To determine prevalence and intensity of pain after stroke, focusing on patients' perspectives. METHODS: During a one year period, 416 first-ever stroke patients were included in the population based Lund Stroke Register. After 4 and 16 months (median), 297 patients (98% of survivors) were followed up. Worst pain intensity during the previous 48 hours was assessed on a visual analogue scale (VAS), range 0 to 100: a score of 0 to 30 was defined as no or mild pain; 40 to 100 as moderate to severe pain. NIH stroke scale (NIHSS) score and HbA1c were assessed at baseline. At 16 months, screening for depression was done using the geriatric depression scale (GDS-20), and cognition with the mini-mental state examination (MMSE). Predictors of pain were determined by multivariate analyses. RESULTS: Moderate to severe pain was reported by 96 patients (32%) after four months (VAS median=60). Predictors of pain were younger age (p=0.01), female sex (p=0.006), higher NIHSS score (p<0.001), and raised HbA1c (p=0.001) at stroke onset. At 16 months, only 62 patients (21%) had moderate to severe pain, but pain intensity was more severe (median VAS score=70; p<0.016). Higher pain intensity correlated with female sex, worse GDS-20 score, better MMSE score, and raised HbA1c. Pain was persistent in 47%, disturbed sleep in 58%, and required rest for relief in 40% of patients. CONCLUSIONS: Although prevalence of pain after stroke decreased with time, after 16 months 21% had moderate to severe pain. Late pain after stroke was on average more severe, and profoundly affected the patients' wellbeing. PMID- 16354738 TI - Stress induced hypotension in pure autonomic failure. AB - A 47 year old woman with pure autonomic failure complained of dizziness during emotional stress. Emotional stimuli have not previously been reported to cause hypotension in patients with autonomic failure. In the patient, ambulatory blood pressure recording revealed severe hypotension (50/30 mm Hg) after a stressful event. During a tilt table test, hyperventilation was shown to cause a significant fall of blood pressure. This suggests that emotional stress can induce hypotension, probably through hyperventilation, in subjects with autonomic failure. PMID- 16354739 TI - Sneeze related area in the medulla: localisation of the human sneezing centre? AB - Sneezing is a rarely explored symptom in neurological practice. In the cat, a sneeze evoking centre is located in the medulla. The existence of a sneezing centre has not been confirmed in humans. A case with abnormal sneezing secondary to a strategic infarct in the right latero-medullary region is presented. A 66 year old man suddenly presented paroxysmal sneezing followed by ataxia, right sided motor and sensory symptoms, and hoarseness. The application of stimuli to the right nasal fossa did not evoke sneezing nor the wish to sneeze. The same stimuli to the contralateral nasal fossa evoked normal sneezing. The preservation of the superficial sensitivity of the nasal fossa indicates that the lesion was localised in the hypothetical human sneezing centre, very close to the spinal trigeminal tract and nucleus. This centre appears to be bilateral and functionally independent on both sides. PMID- 16354740 TI - Workplace protection of air-fed visors used in paint spraying operations. AB - Air-fed visors are commonly used for protection against exposure to airborne isocyanates during paint spraying. Protection levels for this class of equipment are theoretically adequate, yet isocyanate sensitization in this occupation still occurs. The work reported here set out to establish the level of respiratory protection that is achieved during real paint spraying activities when air-fed visors are used. The work also examined the effects of reduced air supply flow rates on this type of respiratory protection. The workplace study highlighted common problems that occur when attempting to measure protection factors, and process and interpret the collected data. Many of the environments included in this study did not exhibit challenge concentrations high enough to reliably measure the workplace protection factor of this class of device. When detection limits are taken into consideration, the remaining field data suggest that an assigned protection factor in the region of 40 may be appropriate. When well maintained and used in accordance with the manufacturer's instructions, air-fed visors are capable of providing a good level of respiratory protection. The protection given by air-fed visors is strongly dependent on the air flow supplied to them. Laboratory measurements demonstrate that protection falls as the air supply falls. This is a gradual process and does not suddenly occur at any particular air supply flow. Observations made during the field tests indicate that there may be other activities associated with the spraying process that need to be taken into consideration when looking for sources of respiratory sensitization. PMID- 16354741 TI - Characterization of microbial particle release from biomass and building material surfaces for inhalation exposure risk assessment. AB - A conceptual approach including measurements of materials at rest (step 1), measurements using a large rotating drum (step 2) or a Particle-FLEC (step 2) and measurements at a workplace (step 4) has been used to characterize the release of microbial components (bacteria, fungi, actinomycetes, endotoxin or enzymes) and particles from straw, wood chips or fungal cultures of different ages on gypsum boards. Repeated agitation or handling periods were included in step 2 and step 4. There was a low similarity between the amount of microbial components measured in step 1 and the aerosolized amount (step 2) from gypsum boards, wood chips and straw. Ratios between some microbial components measured at the workplace (step 4) and measured in step 2, showed similarities. Less than 1.3% of the total amount of microorganisms and endotoxin becomes airborne during 5 min of agitation of straw or wood chips. Most microbial components were released at higher rates during the first agitation period than during the following periods. However, differences were seen between different microbial components, and endotoxin from straw was released at the same rate in two successive agitation periods. Fungal particles smaller than spores were released from fungal colonized gypsum boards at amounts that were up to 30 times higher in the first agitation period compared with that in the following period, while fungal spores were released at amounts that were five times as high in the first period compared with that in the following period. In addition to differences between microbial components, the release patterns of microbial components were different for wood chips and straw. The time for maximum particle release to half particle release was longer for straw than for wood chips. The observation that some components, e.g. endotoxin, are released at the same rate in two successive handling steps, and that others (e.g. fungi) are mainly released initially, shows that the exposure period to different components from the same material differs in duration. The observed differences in the release patterns of different components and the differences between materials are important when preventive steps are to be taken, and it stresses the importance of applying a relevant sampling time and period in exposure assessments. PMID- 16354742 TI - Effect of air pollution on olfactory function in residents of Mexico City. AB - To our knowledge there has been no study of the effect of everyday air pollution on olfactory function. It was therefore the aim of this study to compare the olfactory performance of long-term residents of Mexico City, an environment with high air pollution, with the olfactory performance of residents of the Mexican state of Tlaxcala, a region geographically similar to Mexico City but with low air pollution. Healthy volunteers [82 Mexico City subjects (MEX), 86 Tlaxcala subjects (TLX)] 20-63 years of age and balanced for age and gender between the two localities were tested for the perception of the odors of everyday beverages presented in squeeze bottles. When tested with ascending concentrations of stimuli in a three-way oddball paradigm, residents of Tlaxcala detected the odors of instant coffee and of an orange drink at significantly lower concentrations than residents of Mexico City. They also performed significantly better in discriminating between the two similar-smelling Mexican beverages horchata and atole in an oddball test. Significant differences between the two populations in overall olfactory performance were apparent in three of the four age classes (20- to 29-, 30- to 39-, and 40- to 49-year-old subjects) but not in the 50-63 years age class. About 10% of MEX subjects compared to about 2% of TLX subjects were judged to have poor olfactory function; all were from the older age classes (mean age: 48.6 years). Thus, air pollution in Mexico City appears to have a substantial impact on olfactory function even in young and middle-aged residents. PMID- 16354743 TI - Sniffing and spatiotemporal coding in olfaction. AB - The act of sniffing increases the air velocity and changes the duration of airflow in the nose. It is not yet clear how these changes interact with the intrinsic timing within the olfactory bulb, but this is a matter of current research activity. An action of sniffing in generating a high velocity that alters the sorption of odorants onto the lining of the nasal cavity is expected from the established work on odorant properties and sorption in the frog nose. Recent work indicates that the receptor properties in the olfactory epithelium and olfactory bulb are correlated with the receptor gene expression zones. The responses in both the epithelium and the olfactory bulb are predictable to a considerable extent by the hydrophobicity of odorants. Furthermore, receptor expression in both rodent and salamander nose interacts with the shapes of the nasal cavity to place the receptor sensitivity to odorants in optimal places according to the aerodynamic properties of the nose. PMID- 16354744 TI - Numerical modeling of turbulent and laminar airflow and odorant transport during sniffing in the human and rat nose. AB - Human sniffing behavior usually involves bouts of short, high flow rate inhalation (>300 ml/s through each nostril) with mostly turbulent airflow. This has often been characterized as a factor enabling higher amounts of odorant to deposit onto olfactory mucosa than for laminar airflow and thereby aid in olfactory detection. Using computational fluid dynamics human nasal cavity models, however, we found essentially no difference in predicted olfactory odorant flux (g/cm2 s) for turbulent versus laminar flow for total nasal flow rates between 300 and 1000 ml/s and for odorants of quite different mucosal solubility. This lack of difference was shown to be due to the much higher resistance to lateral odorant mass transport in the mucosal nasal airway wall than in the air phase. The simulation also revealed that the increase in airflow rate during sniffing can increase odorant uptake flux to the nasal/olfactory mucosa but lower the cumulative total uptake in the olfactory region when the inspired air/odorant volume was held fixed, which is consistent with the observation that sniff duration may be more important than sniff strength for optimizing olfactory detection. In contrast, in rats, sniffing involves high frequency bouts of both inhalation and exhalation with laminar airflow. In rat nose odorant uptake simulations, it was observed that odorant deposition was highly dependent on solubility and correlated with the locations of different types of receptors. PMID- 16354745 TI - Activity of mecillinam against ESBL producers in vitro. PMID- 16354746 TI - Effect of 1-(1-naphthylmethyl)-piperazine, a novel putative efflux pump inhibitor, on antimicrobial drug susceptibility in clinical isolates of Enterobacteriaceae other than Escherichia coli. AB - OBJECTIVES: 1-(1-Naphthylmethyl)-piperazine (NMP) has been shown to reverse multidrug resistance (MDR) in Escherichia coli overexpressing RND type efflux pumps, but there is no data on its activity in Enterobacteriaceae other than E. coli. METHODS: The antimicrobial susceptibilities of laboratory strains and 167 clinical isolates of Enterobacteriaceae to a variety of antimicrobial agents were determined in the absence and presence of NMP and, for comparison, of Phe-Arg beta-naphthylamide (PAbetaN), another putative efflux pump inhibitor (EPI). A 4 fold or greater reduction of the MIC after EPI addition was considered significant. RESULTS: NMP consistently reduced the MIC of linezolid in Citrobacter freundii, Enterobacter aerogenes and Klebsiella pneumoniae clinical isolates. Significant effects of NMP addition in >50% of tested isolates were also seen for levofloxacin, tetracycline and chloramphenicol in E. aerogenes, and for levofloxacin and tetracycline in K. pneumoniae, whereas no or minor effects were observed in Serratia marcescens. MDR reversal by NMP was more likely in isolates with decreased susceptibility to fluoroquinolones. In most fluoroquinolone-resistant strains the activity was sufficient to render isolates drug-susceptible at clinically achievable concentrations. The activity of PAbetaN was different from that of NMP, suggesting different modes of action of the two putative EPIs. CONCLUSION: NMP has moderate activity in reversing MDR in many but not all members of the Enterobacteriaceae family including clinical isolates. Its effects on resistance reversal depend on bacterial species and drug, and are different from those seen with PAbetaN. PMID- 16354747 TI - Effect of 1-(1-naphthylmethyl)-piperazine, a novel putative efflux pump inhibitor, on antimicrobial drug susceptibility in clinical isolates of Escherichia coli. AB - OBJECTIVES: 1-(1-Naphthylmethyl)-piperazine (NMP) has been shown to reverse multidrug resistance (MDR) in Escherichia coli overexpressing resistance nodulation-cell division type efflux pumps, but there is no data on its activity in clinical isolates of E. coli. METHODS: The antimicrobial susceptibility of 60 clinical isolates of E. coli to a variety of antimicrobial agents was determined in the absence and presence of NMP and, for comparison, of Phe-Arg-beta naphthylamide (PAbetaN), another putative efflux pump inhibitor (EPI). The intracellular accumulation of ethidium bromide was measured to confirm efflux pump inhibition as the likely mechanism of action of NMP. RESULTS: Based on a 4 fold or greater reduction of the MIC after the addition of NMP in >50% of the isolates, significant effects of NMP at a concentration of 100 mg/L were seen for levofloxacin, linezolid and ethidium bromide. The ethidium bromide MIC changes after NMP addition correlated with differences in the ethidium bromide intracellular accumulation as measured by fluorometry in whole cell accumulation experiments. The activity of PAbetaN was different from that of NMP, in particular regarding macrolide resistance reversal, suggesting different modes of action of the two putative EPIs. CONCLUSIONS: NMP is moderately active in reversing MDR in clinical isolates of E. coli and can partially restore fluoroquinolone susceptibility through inhibition of efflux pumps. PMID- 16354748 TI - Immune reconstitution inflammatory syndrome: more answers, more questions. AB - The institution of highly active antiretroviral therapy (HAART) in HIV-infected patients restores protective immune responses against a wide variety of pathogens and dramatically decreases mortality. In a subset of patients receiving HAART, immune reconstitution is associated with a pathological inflammatory response leading to substantial short-term morbidity and even mortality. The past several years have seen marked advances in our clinical understanding of the immune reconstitution inflammatory syndrome (IRIS), but many questions remain. This article summarizes recent data on clinical risk factors for the development of IRIS. A consistent finding from multiple groups is that IRIS develops in a substantial percentage of HIV-infected patients who have an underlying opportunistic infection and receive HAART. As the use of HAART stands to markedly increase over the next several years, optimal care of patients receiving HAART will need to incorporate monitoring for and treating complications of IRIS. PMID- 16354749 TI - The pathogenesis of obstructive sleep apnea: advances in the past 100 years. PMID- 16354750 TI - Chromosomal mapping of 170 BAC clones in the ascidian Ciona intestinalis. AB - The draft genome ( approximately 160 Mb) of the urochordate ascidian Ciona intestinalis has been sequenced by the whole-genome shotgun method and should provide important insights into the origin and evolution of chordates as well as vertebrates. However, because this genomic data has not yet been mapped onto chromosomes, important biological questions including regulation of gene expression at the genome-wide level cannot yet be addressed. Here, we report the molecular cytogenetic characterization of all 14 pairs of C. intestinalis chromosomes, as well as initial large-scale mapping of genomic sequences onto chromosomes by fluorescent in situ hybridization (FISH). Two-color FISH using 170 bacterial artificial chromosome (BAC) clones and construction of joined scaffolds using paired BAC end sequences allowed for mapping of up to 65% of the deduced 117-Mb nonrepetitive sequence onto chromosomes. This map lays the foundation for future studies of the protochordate C. intestinalis genome at the chromosomal level. PMID- 16354751 TI - Mutation hot spots in mammalian mitochondrial DNA. AB - Animal mitochondrial DNA is characterized by a remarkably high level of within species homoplasy, that is, phylogenetic incongruence between sites of the molecule. Several investigators have invoked recombination to explain it, challenging the dogma of maternal, clonal mitochondrial inheritance in animals. Alternatively, a high level of homoplasy could be explained by the existence of mutation hot spots. By using an exhaustive mammalian data set, we test the hot spot hypothesis by comparing patterns of site-specific polymorphism and divergence in several groups of closely related species, including hominids. We detect significant co-occurrence of synonymous polymorphisms among closely related species in various mammalian groups, and a correlation between the site specific levels of variability within humans (on one hand) and between Hominoidea species (on the other hand), indicating that mutation hot spots actually exist in mammalian mitochondrial coding regions. The whole data, however, cannot be explained by a simple mutation hot spots model. Rather, we show that the site specific mutation rate quickly varies in time, so that the same sites are not hypermutable in distinct lineages. This study provides a plausible mutation model that potentially accounts for the peculiar distribution of mitochondrial sequence variation in mammals without the need for invoking recombination. It also gives hints about the proximal causes of mitochondrial site-specific hypermutability in humans. PMID- 16354752 TI - Logistic regression protects against population structure in genetic association studies. AB - We conduct an extensive simulation study to compare the merits of several methods for using null (unlinked) markers to protect against false positives due to cryptic substructure in population-based genetic association studies. The more sophisticated "structured association" methods perform well but are computationally demanding and rely on estimating the correct number of subpopulations. The simple and fast "genomic control" approach can lose power in certain scenarios. We find that procedures based on logistic regression that are flexible, computationally fast, and easy to implement also provide good protection against the effects of cryptic substructure, even though they do not explicitly model the population structure. PMID- 16354753 TI - Functional genomics of membrane transporters in human populations. AB - Although considerable progress has been made toward characterizing human DNA sequence variation, there remains a deficiency in information on human phenotypic variation at the single-gene level. We systematically analyzed the function of all protein-altering variants of eleven membrane transporters in heterologous expression systems. Coding-region variants were identified by screening DNA from a large sample (n = 247-276) of ethnically diverse subjects. In total, we functionally analyzed 88 protein-altering variants. Fourteen percent of the polymorphic variants (defined as variants with allele frequencies > or =1% in at least one major ethnic group) had no activity or significantly reduced function. Decreased function variants had significantly lower allele frequencies and were more likely to alter evolutionarily conserved amino acid residues. However, variants at evolutionarily conserved positions with approximately normal activity in cellular assays were also at significantly lower allele frequencies, suggesting that some variants with apparently normal activity in biochemical assays may influence occult functions or quantitative degrees of function that are important in human fitness but not measured in these assays. For example, eight (14%) of the 58 variants for which we had measured the transport of at least two substrates showed substrate-specific defects in transport. These variants and the reduced function variants provide plausible candidates for disease susceptibility or variation in clinical drug response. PMID- 16354754 TI - Identification of transposable elements using multiple alignments of related genomes. AB - Accurate genome-wide cataloging of transposable elements (TEs) will facilitate our understanding of mobile DNA evolution, expose the genomic effects of TEs on the host genome, and improve the quality of assembled genomes. Using the availability of several nearly complete Drosophila genomes and developments in whole genome alignment methods, we introduce a large-scale comparative method for identifying repetitive mobile DNA regions. These regions are highly enriched for transposable elements. Our method has two main features distinguishing it from other repeat-finding methods. First, rather than relying on sequence similarity to determine the location of repeats, the genomic artifacts of the transposition mechanism itself are systematically tracked in the context of multiple alignments. Second, we can derive bounds on the age of each repeat instance based on the phylogenetic species tree. We report results obtained using both complete and draft sequences of four closely related Drosophila genomes and validate our results with manually curated TE annotations in the Drosophila melanogaster euchromatin. We show the utility of our findings in exploring both transposable elements and their host genomes: In the study of TEs, we offer predictions for novel families, annotate new insertions of known families, and show data that support the hypothesis that all known TE families in D. melanogaster were recently active; in the study of the host, we show how our findings can be used to determine shifts in the eu-heterochromatin junction in the pericentric chromosome regions. PMID- 16354755 TI - Retrotransposon accumulation and satellite amplification mediated by segmental duplication facilitate centromere expansion in rice. AB - The abundance of repetitive DNA varies greatly across centromeres within an individual or between different organisms. To shed light on the molecular mechanisms of centromere repeat proliferation, we performed structural analysis of LTR-retrotransposons, mostly centromere retrotransposons of rice (CRRs), and phylogenetic analysis of CentO satellite repeats harbored in the core region of the rice chromosome 4 centromere (CEN4). The data obtained demonstrate that the CRRs in the centromeric region we investigated have been enriched more significantly by recent rounds of segmental duplication than by original integration of active elements, suggesting that segmental duplication is an important process for CRR accumulation in the centromeric region. Our results also indicate that segmental duplication of large arrays of satellite repeats is primarily responsible for the amplification of satellite repeats, contributing to rapid reshuffling of CentO satellites. Intercentromere satellite homogenization was revealed by genome-wide comparison of CentO satellite monomers. However, a 10 bp duplication present in nearly half of the CEN4 monomers was found to be completely absent in rice centromere 8 (CEN8), suggesting that CEN4 and CEN8 may represent two different stages in the evolution of rice centromeres. These observations, obtained from the only complex eukaryotic centromeres to have been completely sequenced thus far, depict the evolutionary dynamics of rice centromeres with respect to the nature, timing, and process of centromeric repeat amplification. PMID- 16354756 TI - Modest dietary K+ restriction provokes insulin resistance of cellular K+ uptake and phosphorylation of renal outer medulla K+ channel without fall in plasma K+ concentration. AB - Extracellular K(+) concentration ([K(+)]) is closely regulated by the concerted regulatory responses of kidney and muscle. In this study, we aimed to define the responses activated when dietary K(+) was moderately reduced from a control diet (1.0% K(+)) to a 0.33% K(+) diet for 15 days. Although body weight and baseline plasma [K(+)] (4.0 mM) were not reduced in the 0.33% K(+) group, regulatory responses to conserve plasma [K(+)] were evident in both muscle and kidney. Insulin-stimulated clearance of K(+) from the plasma was estimated in vivo in conscious rats with the use of tail venous and arterial cannulas. During infusion of insulin.(50 mU.kg(-1).min(-1)), plasma [K(+)] level fell to 3.2 +/- 0.1 mM in the 1.0% K(+) diet group and to only 3.47 +/- 0.07 mM in the 0.33% K(+) diet group (P < 0.01) with no reduction in urinary K(+) excretion, which is evidence of insulin resistance to cellular K(+) uptake. Insulin-stimulated cellular K(+) uptake was quantitated by measuring the K(+) infusion rate necessary to clamp plasma K(+) at baseline (in micromol.kg(-1).min(-1)) during 5 mU of insulin.kg( 1).min(-1) infusion: 9.7 +/- 1.5 in 1% K(+) diet was blunted to 5.2 +/- 1.7 in the 0.33% K(+) diet group (P < 0.001). Muscle [K(+)] and Na(+)-K(+)-ATPase activity and abundance were unchanged during the 0.33% K(+) diet. Renal excretion, which was measured overnight in metabolic cages, was reduced by 80%, from 117.6 +/- 10.5 micromol/h/animal (1% K(+) diet) to 24.2 +/- 1.7 micromol/h/animal (0.33% K(+) diet) (P < 0.001). There was no significant change in total abundance of key renal K(+) transporters, but 50% increases in both renal PTK cSrc abundance and ROMK phosphorylation in the 0.33% K(+) vs. 1% K(+) diet group, previously established to be associated with internalization of ROMK. These results indicate that plasma [K(+)] can be maintained during modest K(+) restriction due to a decrease in insulin-stimulated cellular K(+) uptake as well as renal K(+) conservation mediated by inactivation of ROMK, both without a detectable change in plasma [K(+)]. The error signals inciting and maintaining these responses remain to be identified. PMID- 16354757 TI - Induced focal adhesion kinase expression suppresses apoptosis by activating NF kappaB signaling in intestinal epithelial cells. AB - Focal adhesion kinase (FAK) integrates various extracellular and intracellular signals and is implicated in a variety of biological functions, but its exact role and downstream targeting signals in the regulation of apoptosis in intestinal epithelial cells (IECs) remains unclear. The current study tested the hypothesis that FAK has an antiapoptotic role in the IEC-6 cell line by altering NF-kappaB signaling. Induced FAK expression by stable transfection with the wild type (WT)-FAK gene increased FAK phosphorylation, which was associated with an increase in NF-kappaB activity. These stable WT-FAK-transfected IECs also exhibited increased resistance to apoptosis when they were exposed to TNF-alpha plus cycloheximide (TNF-alpha/CHX). Specific inhibition of NF-kappaB by the recombinant adenoviral vector containing the IkappaBalpha superrepressor prevented increased resistance to apoptosis in WT-FAK-transfected cells. In contrast, inactivation of FAK by ectopic expression of dominant-negative mutant of FAK (DNM-FAK) inhibited NF-kappaB activity and increased the sensitivity to TNF-alpha/CHX-induced apoptosis. Furthermore, induced expression of endogenous FAK by depletion of cellular polyamines increased NF-kappaB activity and resulted in increased resistance to TNF-alpha/CHX-induced apoptosis, both of which were prevented by overexpression of DNM-FAK. These results indicate that increased expression of FAK suppresses TNF-alpha/CHX-induced apoptosis, at least partially, through the activation of NF-kappaB signaling in IECs. PMID- 16354758 TI - Nitric oxide attenuates IGF-I-induced aortic smooth muscle cell motility by decreasing Rac1 activity: essential role of PTP-PEST and p130cas. AB - Recent data support the hypothesis that reactive oxygen species (ROS) play a central role in the initiation and progression of vascular diseases. An important vasoprotective function related to the regulation of ROS levels appears to be the antioxidant capacity of nitric oxide (NO). We previously reported that treatment with NO decreases phosphotyrosine levels of adapter protein p130(cas) by increasing protein tyrosine phosphatase-proline, glutamate, serine, and threonine sequence protein (PTP-PEST) activity, which leads to the suppression of agonist induced H(2)O(2) elevation and motility in cultured rat aortic smooth muscle cells (SMCs). The present study was performed to investigate the hypotheses that 1) IGF-I increases the activity of the small GTPase Rac1 as well as H(2)O(2) levels and 2) NO suppresses IGF-I-induced H(2)O(2) elevation by decreasing Rac1 activity via increased PTP-PEST activity and dephosphorylation of p130(cas). We report that IGF-I induces phosphorylation of p130(cas) and activation of Rac1 and that NO attenuates these effects. The effects of NO are mimicked by the overexpression of PTP-PEST or dominant-negative (dn)-p130(cas) and antagonized by the expression of dn-PTP-PEST or p130(cas). We conclude that IGF-I induces rat aortic SMC motility by increasing phosphotyrosine levels of p130(cas) and activating Rac1 and that NO decreases motility by activating PTP-PEST, inducing dephosphorylating p130(cas), and decreasing Rac1 activity. Decreased Rac1 activity lowers intracellular H(2)O(2) levels, thus attenuating cell motility. PMID- 16354759 TI - Commentary: seeing the light. PMID- 16354761 TI - A refinement to 'how many genes underlie the occurrence of common complex diseases in the population?'. PMID- 16354762 TI - How batrachotoxin modifies the sodium channel permeation pathway: computer modeling and site-directed mutagenesis. AB - A structural model of the rNav1.4 Na+ channel with batrachotoxin (BTX) bound within the inner cavity suggested that the BTX pyrrole moiety is located between a lysine residue at the DEKA selectivity filter (Lys1237) and an adjacent phenylalanine residue (Phe1236). We tested this pyrrole-binding model by site directed mutagenesis of Phe1236 at D3/P-loop with 11 amino acids. Mutants F1236D and F1236E expressed poorly, whereas nine other mutants either expressed robust Na+ currents, like the wild-type (F1236Y/Q/K), or somewhat reduced current (F1236G/A/C/N/W/R). Gating properties were altered modestly in most mutant channels, with F1236G displaying the greatest shift in activation and steady state fast inactivation (-10.1 and -7.5 mV, respectively). Mutants F1236K and F1236R were severely resistant to BTX after 1000 repetitive pulses (+50 mV/20 ms at 2 Hz), whereas seven other mutants were sensitive but with reduced magnitudes compared with the wild type. It is noteworthy that rNav1.4-F1236K mutant Na+ channels remained highly sensitive to block by the local anesthetic bupivacaine, unlike several other BTX-resistant mutant channels. Our data thus support a model in which BTX, when bound within the inner cavity, interacts with the D3/P-loop directly. Such a direct interaction provides clues on how BTX alters the Na+ channel selectivity and conductance. PMID- 16354763 TI - Angiotensin II up-regulates the leukemia-associated Rho guanine nucleotide exchange factor (RhoGEF), a regulator of G protein signaling domain-containing RhoGEF, in vascular smooth muscle cells. AB - In vascular smooth muscle, stimulation of heterotrimeric G protein-coupled receptors (GPCRs) by various contractile agonists activates intracellular signaling molecules to result in an increase in cytosolic Ca2+ and the subsequent phosphorylation of myosin light chain (MLC) by Ca2+/calmodulin-dependent MLC kinase. In addition, a portion of agonist-induced contraction is partially mediated by the Ca2+-independent activation of the small G protein RhoA and a downstream target, Rho-kinase. The activation of RhoA is controlled by several regulatory proteins, including guanine nucleotide exchange factors (GEFs). GEFs activate RhoA by promoting the release of GDP and then facilitating the binding of GTP. There are three Rho-specific GEFs (RhoGEFs) in vascular smooth muscle that contain a binding domain [regulator of G protein signaling (RGS) domain] capable of linking GPCRs to RhoA activation: PDZ-RhoGEF, leukemia-associated RhoGEF (LARG), and p115RhoGEF. We hypothesized that RGS domain-containing RhoGEFs, especially LARG, participate in linking GPCR to RhoA activation in vascular smooth muscle. We observed that angiotensin II up-regulates LARG via the AT1 receptor, and this up-regulation is signaled via the phosphatidylinositol 3 kinase pathway. Furthermore, angiotensin II treatment caused a small, but significant, increase in the component of contractile responses sensitive to Rho kinase antagonism. These observations support the hypothesis that RhoGEFs, particularly LARG, participate in linking GPCR to RhoA activation in vascular smooth muscle. PMID- 16354764 TI - Glabridin suppresses intercellular adhesion molecule-1 expression in tumor necrosis factor-alpha-stimulated human umbilical vein endothelial cells by blocking sphingosine kinase pathway: implications of Akt, extracellular signal regulated kinase, and nuclear factor-kappaB/Rel signaling pathways. AB - (R)-4-(3,4-Dihydro-8,8-dimethyl)-2H,8H-benzo[1,2-b:3,4-b'] dipyran-3yl)-1,3 benzenediol (glabridin) is known to have anti-inflammatory, antimicrobial, and cardiovascular protective activities. In the present study, we report the inhibitory effect of glabridin on intercellular adhesion molecule-1 (ICAM-1) expression in tumor necrosis factor-alpha (TNF-alpha)-stimulated human umbilical vein endothelial cells (HUVECs). Glabridin inhibited THP-1 cell adhesion to HUVECs stimulated by TNF-alpha and cell surface expression of ICAM-1 in TNF-alpha stimulated HUVECs. The mRNA expression of adhesion molecules, including ICAM-1, vascular cell adhesion molecule-1, and E-selectin, was also suppressed by glabridin. Further study demonstrated the inhibitory effect of glabridin on nuclear factor (NF)-kappaB/Rel DNA binding, inhibitory factor-kappaB alpha (IkappaB alpha), and IkappaB beta degradation, IkappaB kinase activation, and p65 nuclear translocation in TNF-alpha-stimulated HUVECs. Treatment of a variety of cell lines with glabridin revealed that inhibitory effect of glabridin on NF kappaB/Rel activation is not cell type-specific, and both inducible and constitutive NF-kappaB/Rel activation was suppressed by glabridin treatment. Moreover, TNF-alpha-induced phosphorylation of Akt and extracellular signal regulated kinase (ERK) was blocked by glabridin treatment in HUVECs. Glabridin also suppressed sphingosine-1-phosphate (S1P)-induced cell surface expression and mRNA expression of ICAM-1. Further study demonstrated that TNF-alpha-induced sphingosine kinase activity was inhibited by glabridin, and the inhibitory effect of glabridin on TNF-alpha-induced ICAM-1 expression was reversed by addition of exogenous S1P. Together, our results indicate that the inhibitory effect of glabridin on ICAM-1 expression might be mediated, at least in part, by inhibiting sphingosine kinase pathway and subsequent inhibition of signaling pathways, including Akt, ERK, and NF-kappaB/Rel signaling pathway. PMID- 16354765 TI - The teratogenic sensitivity to 2,3,7,8-tetrachlorodibenzo-p-dioxin is modified by a locus on mouse chromosome 3. AB - In an effort to understand how genetics can influence individual sensitivity to environmentally induced disease, we performed a linkage analysis to identify murine loci in addition to the Ahr locus that influence the incidence of cleft palate and hydronephrosis in developing mice exposed to the pollutant 2,3,7,8 tetrachlorodibenzo-p-dioxin (dioxin). Administration of 64 microg/kg dioxin to C57BL/6J (B6) dams at embryonic day 9 (E9) led to palatal clefting and hydronephrosis in nearly 100% of embryos by E17. In contrast, similar exposure of CBA/J (CBA) dams led to cleft palate in only 8% and hydronephrosis in 69% of embryos. To determine the genetic basis for this strain-dependent sensitivity, linkage analyses on the progeny of a B6CBAF1 intercross and a CBAxB6CBAF1 backcross were performed. The incidences of cleft palate and hydronephrosis were assessed and genomic DNA from embryos was analyzed at informative simple sequence length polymorphism (SSLP) markers. One locus segregating with dioxin-induced cleft palate was identified (p < 0.01) and designated as chemically mediated teratogenesis number 1 (Cmt1). The Cmt1 locus is located on chromosome 3. PMID- 16354766 TI - Novel targeting of cyclooxygenase-2 (COX-2) pre-mRNA using antisense morpholino oligonucleotides directed to the 3' acceptor and 5' donor splice sites of exon 4: suppression of COX-2 activity in human amnion-derived WISH and myometrial cells. AB - Increased expression of cyclooxygenase-2 (COX-2) has been implicated in the onset of both term and preterm labor. In this context, both selective and nonselective COX-2 inhibitors have been used in clinical trials to determine their efficacy in delaying preterm labor. However, recent evidence indicates that these tocolytics may have potentially adverse fetal and maternal side effects. Therefore, the development of more specific and nontoxic agents to inhibit COX-2 needs to be considered. We have evaluated whether antisense morpholino oligonucleotides have therapeutic potential in inhibiting COX-2 by specifically targeting both the 3' and 5' acceptor and donor sites of exon 4 of COX-2's pre-mRNA sequence. Confocal microscopy on "live" cells illustrated high levels of penetrance of antisense morpholino oligonucleotides using the Endo-Porter formula (Gene-Tools, LLC, Philomath, OR), with delivery efficiencies of 82 and 78%, respectively, in amnion derived WISH and myometrial cells. Substantial inhibition by the morpholino oligonucleotides of COX-2 expression, induced by lipopolysaccharide administration, was observed at both the mRNA and protein levels. Loss of enzymic activity of COX-2 was confirmed using a sensitive COX enzyme activity assay, which reflects the rate of conversion of arachidonic acid to prostaglandin H2. Our results indicate that antisense morpholino oligonucleotides significantly inhibit expression and activity of this enzyme in in vitro cultures of amnion WISH and myometrial cells. The potential thus exists that a similar approach can be mimicked in vivo to produce a highly specific and nontoxic strategy to inhibit COX-2 activity with its subsequent effects on the better management of preterm labor and other inflammatory conditions. PMID- 16354767 TI - Zinc and mercuric ions distinguish TRESK from the other two-pore-domain K+ channels. AB - TWIK-related spinal cord K+ channel (TRESK) is the most recently cloned two-pore domain potassium (2PK+) channel, regulated by the calcium/calmodulin-dependent protein phosphatase calcineurin. Functional identification of endogenous TRESK and its distinction from the other 2PK+ channels, producing similar background K+ current, are impeded by the lack of specific inhibitors. Therefore, we searched for antagonists selective against TRESK among the mouse 2PK+ channels by screening more than 200 substances. Mibefradil, zinc, and mercuric ions inhibited TRESK expressed in Xenopus laevis oocytes with IC50 values lower than 10 microM. The specificity of the identified agents was determined by measuring their effects on mouse TALK-1, TASK-1, TASK-2, TASK-3, THIK-1, TRAAK, TREK-1, and TREK 2. Mibefradil failed to discriminate well among the functional 2PK+ channels; however, Zn2+ and Hg2+ exerted a significantly stronger inhibitory effect on TRESK than on the other channels. Sensitivity to zinc but insensitivity to ruthenium red were distinctive features of TRESK. Whereas both Zn2+ and Hg2+ were selective blockers of TRESK among the mouse 2PK+ channels, human TRESK was resistant to Zn2+; it was blocked only by Hg2+. His132 of mouse TRESK was partly responsible for this difference. Mouse TRESK expressed in COS-7 cells was also inhibited by Zn2+ and Hg2+, and TRESK single-channel current was diminished in outside-out patches, indicating that the action of the ions was membrane delimited, most probably targeting the channel itself. Thus, both Zn2+ and Hg2+ are expected to inhibit endogenous TRESK in isolated mouse cells, and these ions can be applied to identify the calcineurin-activated 2PK+ channel in its natural environment. PMID- 16354768 TI - Janus kinase-signal transducer and activator of transcription mediates phosphatidic acid-induced interleukin (IL)-1beta and IL-6 production. AB - We have found previously that phosphatidic acid (PA) can induce inflammatory mediators such as cytokines, which implies that PA plays a role in inflammatory response. In the present study, we provide evidence of the PA-mediated activation of the Janus tyrosine kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway, which results in the production of interleukin (IL)-1beta and IL-6. PA elicited the rapid phosphorylations of JAK2 and STAT1/3, and the subsequent nuclear translocation. Macrophages that had been transiently transfected with a luciferase reporter construct containing eight consecutive gamma-interferon activating sequence (GAS) elements, a known STAT binding site, exhibited enhanced reporter gene activity in response to PA stimulation, which further supports the involvement of JAK-STAT activation in the PA-induced signaling pathway. Of the inflammatory cytokines, IL-1beta, IL-6, and tumor necrosis factor (TNF)-alpha were detected in media from macrophages stimulated with PA. Moreover, the JAK2 inhibitor alpha-cyano-(3,4-dihydroxy)-N benzylcinnamide (AG-490) abolished PA-induced IL-1beta and IL-6 release but not TNF-alpha production, which is consistent with the notion that IL-1beta and IL-6 but not TNF-alpha contain a STAT binding element in their promoter region. The knockdown of JAK2 in macrophages by small interfering RNA significantly attenuated PA-induced IL-1beta and IL-6 production. In addition, JAK2 inhibitor suppressed PA-induced Akt phosphorylation, and the Akt inhibitor 2-(4 morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) blocked GAS activation (GAS contains a promoter that responds to PA), suggesting that PA-mediated JAK2 activation leads to phosphatidylinositol 3-kinase/Akt phosphorylation and STAT activation, and the subsequent translocation of STAT to the nucleus. Together, our data demonstrate that PA-activated macrophages produce IL-1beta and IL-6 and that these processes require the activation of the JAK2-STAT1/3 or JAK2-Akt-STAT signaling pathways. PMID- 16354769 TI - Involvement of Nrf2, p38, B-Raf, and nuclear factor-kappaB, but not phosphatidylinositol 3-kinase, in induction of hemeoxygenase-1 by dietary polyphenols. AB - The highly inducible enzyme, hemeoxygenase-1 (HO-1), metabolizes heme, thereby protecting a variety of cells against oxidative stress and apoptosis. Up regulation by cancer chemopreventive agents has been reported, but its regulation and function in transformed cells are unclear. We compared induction by two dietary polyphenols, curcumin and epigallocatechin-3-gallate (EGCG), with that by the endogenous substrate hemin in epithelial and endothelial cells and examined the relevance to apoptosis. Curcumin or hemin (20 microM) induced HO-1 in breast cells from 5 to 24 h. Curcumin (5-40 microM) or hemin (5-100 microM) induced HO-1 and nuclear levels of nuclear factor (erythroid-derived 2)-related factor (Nrf2) in a dose-dependent manner. EGCG had no effect in breast cells, but at 30 microM, it induced nuclear translocation of Nrf2 and HO-1 expression in B-lymphoblasts. In all cases, induction was inhibited by pretreatment with the phosphatidylinositol 3-kinase (PI3K) inhibitor 2-(4-morpholinyl)-8-phenyl-4H-1 benzopyran-4-one (LY294002) or the p38 inhibitor 4-(4-fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580). The nuclear factor kappaB (NF-kappaB)-DNA binding inhibitor helenalin (20 microM) also prevented induction. However, wortmannin had no effect, suggesting that PI3K was not involved. Curcumin and hemin also induced nuclear Nrf2 and HO-1 effectively in wild-type mouse embryo fibroblasts (wt MEFs) and in B-Raf(-/-) MEFs but not in Nrf2(-/-) MEFs. However, EGCG (5-20 microM) induced HO-1 only in wt MEFs. Results suggest that signaling through p38 mitogen-activated protein kinase, NF-kappaB, and Nrf2 as well as other unidentified molecules is involved in HO-1 induction by hemin and both polyphenols, but cell-specific factors also play a role, particularly with respect to EGCG. Induction of HO-1 by curcumin, EGCG, or low concentrations (5-10 microM) of helenalin did not protect MDA-MB468 breast cells or B-lymphoblasts from apoptosis. PMID- 16354771 TI - Iron imports. III. Transfer of iron from the mucosa into circulation. AB - Transfer of iron from the mucosa is a critical step in dietary iron assimilation that is tightly regulated to ensure the appropriate amount of iron is absorbed to meet the body's demands. Too much iron is highly toxic, and failure to properly control intestinal iron export causes iron overload associated with hereditary forms of hemochromatosis. One form of genetic iron overload, ferroportin disease, originates due to defects in ferroportin, the membrane iron exporter. Ferroportin acts in conjunction with the intestinal ferroxidase hephaestin to mediate release of iron from the enterocyte. How iron is then acquired by transferrin and released into circulation remains an unknown step in this process. PMID- 16354773 TI - Calibration of magnetic and celestial compass cues in migratory birds--a review of cue-conflict experiments. AB - Migratory birds use multiple sources of compass information for orientation, including the geomagnetic field, the sun, skylight polarization patterns and star patterns. In this paper we review the results of cue-conflict experiments designed to determine the relative importance of the different compass mechanisms, and how directional information from these compass mechanisms is integrated. We focus on cue-conflict experiments in which the magnetic field was shifted in alignment relative to natural celestial cues. Consistent with the conclusions of earlier authors, our analyses suggest that during the premigratory season, celestial information is given the greatest salience and used to recalibrate the magnetic compass by both juvenile and adult birds. Sunset polarized light patterns from the region of the sky near the horizon appear to provide the calibration reference for the magnetic compass. In contrast, during migration, a majority of experiments suggest that birds rely on the magnetic field as the primary source of compass information and use it to calibrate celestial compass cues, i.e. the relative saliency of magnetic and celestial cues is reversed. An alternative possibility, however, is suggested by several experiments in which birds exposed to a cue conflict during migration appear to have recalibrated the magnetic compass, i.e. their response is similar to that of birds exposed to cue conflicts during the premigratory season. The general pattern to emerge from these analyses is that birds exposed to the cue conflict with a view of the entire sunset sky tended to recalibrate the magnetic compass, regardless of whether the cue conflict occurred during the premigratory or migratory period. In contrast, birds exposed to the cue conflict in orientation funnels and registration cages that restricted their view of the region of sky near the horizon (as was generally the case in experiments carried out during the migratory season) did not recalibrate the magnetic compass but, instead, used the magnetic compass to calibrate the other celestial compass systems. If access to critical celestial cues, rather than the timing of exposure to the cue conflict (i.e. premigratory vs migratory), determines whether recalibration of the magnetic compass occurs, this suggests that under natural conditions there may be a single calibration reference for all of the compass systems of migratory birds that is derived from sunset (and possibly also sunrise) polarized light cues from the region of sky near the horizon. In cue-conflict experiments carried out during the migratory season, there was also an interesting asymmetry in the birds' response to magnetic fields shifted clockwise and counterclockwise relative to celestial cues. We discuss two possible explanations for these differences: (1) lateral asymmetry in the role of the right and left eye in mediating light-dependent magnetic compass orientation and (2) interference from the spectral and intensity distribution of skylight at sunset with the response of the light-dependent magnetic compass. PMID- 16354774 TI - Pupil shapes and lens optics in the eyes of terrestrial vertebrates. AB - Animal eyes that are primarily used under low-light conditions usually have optical systems of short depth of focus, such that chromatic defocus may lead to considerable blurring of the images. In some vertebrates, the problem is solved by multifocal lenses having concentric zones of different focal lengths, each of which focuses a different relevant spectral range onto the retina. A partially constricted circular pupil would shade the peripheral zones of the lens, leading to the loss of well-focused images at relevant wavelengths. The slit pupil, however, allows for use of the full diameter of the lens even in bright light. We studied species of terrestrial vertebrates from a variety of phylogenetic groups to establish how widespread multifocal lenses are and how pupil shapes are adapted to the optical systems. We found that multifocal lenses are common from amphibians to mammals, including primates. Slit pupils were only present in animals having multifocal optical systems. Among the felids, small species have multifocal lenses and slit pupils, while large species have monofocal lenses and round pupils. The Eurasian lynx, a cat of intermediate size, has an intermediate eye design. The functional significance of the absence of multifocal optical systems in large felids remains mysterious, because such systems are present in other large-eyed terrestrial vertebrates. Multifocal optical systems in nocturnal prosimians suggest that those animals have colour vision despite being described as cone monochromats. PMID- 16354775 TI - Ant navigation: resetting the path integrator. AB - Desert ants use path integration as their predominant system of long-distance navigation, but they also make use of route-defining and nest-defining visual landmarks. Such landmark-gained information might override the information provided by the path integrator, but nevertheless the path integrator keeps running. Here we show that only cues that are associated with the ant being inside the nest are able to reset the path integrator to zero state. Ants were captured at a feeder, i.e. without having run off their home vector, and were forced to enter the nest. On their next outbound run their walking direction differed by 180 degrees from that of ants that had also been captured at the feeder but instead of having been forced to enter the interior of the nest were released at its entrance. Whereas these latter ants still ran off their home vector pointing in the feeder-nest direction (and by this departed from the nest in a direction opposite to the feeder direction) the former ants had reset their home vector to zero state, and had therefore been able to reload their learned feeder vector, and consequently departed from the nest in the feeder direction. Owing to its egocentric nature the path-integrator is error prone. Hence, it is a suitable strategy to reset the path integrator if the ant has appeared at its final goal, the nest. Otherwise during consecutive foraging runs navigational errors would steadily increase. PMID- 16354776 TI - Induced airflow in flying insects I. A theoretical model of the induced flow. AB - A strong induced flow structure envelops the body of insects and birds during flight. This flow influences many physiological processes including delivery of odor and mechanical stimuli to the sensory organs, as well as mass flow processes including heat loss and gas exchange in flying animals. With recent advances in near-field aerodynamics of insect and bird flight, it is now possible to determine how wing kinematics affects induced flow over their body. In this paper, I develop a theoretical model based in rotor theory to estimate the mean induced flow over the body of flapping insects. This model is able to capture some key characteristics of mean induced flow over the body of a flying insect. Specifically, it predicts that induced flow is directly proportional to wing beat frequency and stroke amplitude and is also affected by a wing shape dependent parameter. The derivation of induced flow includes the determination of spanwise variation of circulation on flapping wings. These predictions are tested against the available data on the spanwise distribution of aerodynamic circulation along finite Drosophila melanogaster wings and mean flows over the body of Manduca sexta. To explicitly account for tip losses in finite wings, a formula previously proposed by Prandtl for a finite blade propeller system is tentatively included. Thus, the model described in this paper allows us to estimate how far-field flows are influenced by near-field events in flapping flight. PMID- 16354777 TI - Induced airflow in flying insects II. Measurement of induced flow. AB - The flapping wings of insects and birds induce a strong flow over their body during flight. Although this flow influences the sensory biology and physiology of a flying animal, there are very little data on the characteristics of this self-generated flow field or its biological consequences. A model proposed in the companion paper estimated the induced flow over flying insects. In this study, we used a pair of hot wire anemometers to measure this flow at two locations near the body of a tethered flapping hawk moth, Manduca sexta. The axial inflow anemometer measured the airflow prior to its entry into the stroke plane, whereas the radial outflow anemometer measured the airflow after it crossed the stroke plane. The high temporal resolution of the hot wire anemometers allowed us to measure not only the mean induced flow but also subtle higher frequency disturbances occurring at 1-4 times the wing beat frequency. These data provide evidence for the predictions of a mathematical model proposed in the companion paper. Specifically, the absolute value of the measured induced flow matches the estimate of the model. Also, as predicted by the model, the induced flow varies linearly with wing beat frequency. Our experiments also show that wing flexion contributes significantly to the observed higher frequency disturbances. Thus, the hot wire anemometry technique provides a useful means to quantify the aerodynamic signature of wing flexion. The phasic and tonic components of induced flow influence several physiological processes such as convective heat loss and gas exchange in endothermic insects, as well as alter the nature of mechanosensory and olfactory stimuli to the sensory organs of a flying insect. PMID- 16354778 TI - Trabecular bone in the bird knee responds with high sensitivity to changes in load orientation. AB - Wolff's law of trajectorial orientation proposes that trabecular struts align with the orientation of dominant compressive loads within a joint. Although widely considered in skeletal biology, Wolff's law has never been experimentally tested while controlling for ontogenetic stage, activity level, and species differences, all factors that may affect trabecular bone growth. Here we report an experimental test of Wolff's law using a within-species design in age-matched subjects experiencing physiologically normal levels of bone strain. Two age matched groups of juvenile guinea fowl Numida meleagris ran on a treadmill set at either 0 degrees (Level group) or 20 degrees (Incline group), for 10 min per day over a 45-day treatment period. Birds running on the 20 degrees inclined treadmill used more-flexed knees than those in the Level group at midstance (the point of peak ground reaction force). This difference in joint posture enabled us to test the sensitivity of trabecular alignment to altered load orientation in the knee. Using a new radon transform-based method for measuring trabecular orientation, our analysis shows that the fine trabecular bone in the distal femur has a high degree of correspondence between changes in joint angle and trabecular orientation. The sensitivity of this response supports the prediction that trabecular bone adapts dynamically to the orientation of peak compressive forces. PMID- 16354779 TI - Take-off and landing forces in jumping frogs. AB - Anurans use a saltatorial (jumping) mode of locomotion. A jumping cycle can be divided into four subphases: propulsion, flight, landing and recovery. We studied the landing phase during locomotion in Rana esculenta by measuring the ground reaction forces during propulsion and landing over a range of distances. Landing performance affects locomotor ability in jumping frogs. Landing and recovery together take up one third of the locomotor cycle. Peak landing forces are on average almost three times larger than propulsive forces. The forelimbs appear to be fully extended when they make contact with the substrate and absorb the first impact peak. The height of this peak varies depending on arm positioning and jumping distance. Since the stiffness of the arms stays constant over the full jumping range, it is possible that this is a limiting factor in the ability of the forelimbs to work as dampers. A spring-dashpot model is used to model the effect of arm angle at touch down. Damping during landing is performed by placing the forelimbs at an optimal angle to cancel frictional forces effectively. PMID- 16354780 TI - Packaging of chemicals in the defensive secretory glands of the sea hare Aplysia californica. AB - Sea hares protect themselves from predatory attacks with several modes of chemical defenses. One of these is inking, which is an active release of a protective fluid upon predatory attack. In many sea hares including Aplysia californica and A. dactylomela, this fluid is a mixture of two secretions from two separate glands, usually co-released: ink, a purple fluid from the ink gland; and opaline, a white viscous secretion from the opaline gland. These two secretions are mixed in the mantle cavity and directed toward the attacking predator. Some of the chemicals in these secretions and their mechanism of action have been identified. In our study, we used western blots, immunocytochemistry, amino acid analysis, and bioassays to examine the distribution of these components: (1) an L-amino acid oxidase called escapin for A. californica and dactylomelin-P for A. dactylomela, which has antimicrobial activity but we believe its main function is in defending sea hares against predators that evoke its release; and (2) escapin's major amino acid substrates--L-lysine and L arginine. Escapin is exclusively produced in the ink gland and is not present in any other tissues or secretions. Furthermore, escapin is only sequestered in the amber vesicles of the ink glandand not in the red-purple vesicles, which contain algal-derived chromophores that give ink its distinctive purple color. The concentration of escapin and dactylomelin-P in ink, both in the gland and after its release, is as high as 2 mg ml(-1), or 30 micromol ml(-1), which is well above its antimicrobial threshold. Lysine and arginine (and other amino acids) are packaged into vesicles in the ink and opaline glands, but arginine is present in ink and opaline at <1 mmol l(-1) and lysine is present in ink at <1 mmol l(-1) but in opaline at 65 mmol l(-1). Our previous results showed that both lysine and arginine mediate escapin's bacteriostatic effects, but only lysine mediates its bactericidal effects. Given that escapin's antimicrobial effects require concentrations of lysine and/or arginine >1 mmol l(-1), our data lead us to conclude that lysine in opaline is the primary natural substrate for escapin in ink. Furthermore, packaging of the enzyme escapin and its substrate lysine into two separate glands and their co-release and mixing at the time of predatory attack allows for the generation of bioactive defensive compounds from innocuous precursors at the precise time they are needed. Whether lysine and/or arginine are substrates for escapin's antipredatory functions remains to be determined. PMID- 16354781 TI - Dietary choice behavior in Caenorhabditis elegans. AB - Animals have evolved diverse behaviors that serve the purpose of finding food in the environment. We investigated the food seeking strategy of the soil bacteria eating nematode Caenorhabditis elegans. C. elegans bacterial food varies in quality: some species are easy to eat and support worm growth well, while others do not. We show that worms exhibit dietary choice: they hunt for high quality food and leave hard-to-eat bacteria. This food seeking behavior is enhanced in animals that have already experienced good food. When hunting for good food, worms alternate between two modes of locomotion, known as dwelling: movement with frequent stops and reversals; and roaming: straight rapid movement. On good food, roaming is very rare, while on bad food it is common. Using laser ablations and mutant analysis, we show that the AIY neurons serve to extend roaming periods, and are essential for efficient food seeking. PMID- 16354782 TI - On the importance of radiative heat exchange during nocturnal flight in birds. AB - Many migratory flights take place during cloudless nights, thus under conditions where the sky temperature can commonly be 20 degrees C below local air temperature. The sky then acts as a radiative sink, leading objects exposed to it to have a lower surface temperature than unexposed ones because less infrared energy is received from the sky than from the surfaces that are isothermic to air. To investigate the significance of this effect for heat dissipation during nocturnal flight in birds, we built a wind tunnel with the facility to control wall temperature (TASK) and air temperature (TAIR) independently at air speeds (UWIN) comparable to flying speeds. We used it to measure the influence of TASK, TAIR and UWIN on plumage and skin temperatures in pigeons having to dissipate a thermal load while constrained at rest in a flight posture. Our results show that the temperature of the flight and insulation plumages exposed to a radiative sink can be accurately described by multiple regression models (r2>0.96) based only on TAIR, TASK and UWIN. Predictions based on these models indicate that while convection dominates heat loss for a plumage exposed to air moving at flight speed in a thermally uniform environment, radiation may dominate in the presence of a radiative sink comparable to a clear sky. Our data also indicate that reducing TASK to a temperature 20 degrees C below TAIR can increase the temperature difference across the exposed plumage by at least 13% and thus facilitate heat flow through the main thermal resistance to the loss of internally produced heat in birds. While extrapolation from our experimentally constrained conditions to free flight in the atmosphere is difficult, our results suggest that the sky temperature has been a neglected factor in determining the range of TAIR over which prolonged flight is possible. PMID- 16354783 TI - Maximum aerobic performance in lines of Mus selected for high wheel-running activity: effects of selection, oxygen availability and the mini-muscle phenotype. AB - We compared maximum aerobic capacity during forced exercise (VO2max) in hypoxia (PO2=14% O2), normoxia (21%) and hyperoxia (30%) of lines of house mice selectively bred for high voluntary wheel running (S lines) with their four unselected control (C) lines. We also tested for pleiotropic effects of the ;mighty mini-muscle' allele, a Mendelian recessive that causes a 50% reduction in hind limb muscle but a doubling of mass-specific aerobic enzyme activity, among other pleiotropic effects. VO2max of female mice was measured during forced exercise on a motorized treadmill enclosed in a metabolic chamber that allowed altered PO2. Individual variation in VO2max was highly repeatable within each PO2, and values were also significantly correlated across PO2. Analysis of covariance showed that S mice had higher body-mass-adjusted VO2max than C at all PO2, ranging from +10.7% in hypoxia to +20.8% in hyperoxia. VO2max of S lines increased practically linearly with PO2, whereas that of C lines plateaued from normoxia to hyperoxia, and respiratory exchange ratio (=CO2 production/VO2max) was lower for S lines. These results suggest that the physiological underpinnings of VO2max differ between the S and C lines. Apparently, at least in S lines, peripheral tissues may sustain higher rates of oxidative metabolism if central organs provide more O2. Although the existence of central limitations in S lines cannot be excluded based solely on the present data, we have previously reported that both S and C lines can attain considerably higher VO2max during cold exposure in a He-O2 atmosphere, suggesting that limitations on VO2max depend on interactions between the central and peripheral organs involved. In addition, mini-muscle individuals had higher VO2max than did those with normal muscles, suggesting that the former might have higher hypoxia tolerance. This would imply that the mini-muscle phenotype could be a good model to test how exercise performance and hypoxia tolerance could evolve in a correlated fashion, as previous researchers have suggested. PMID- 16354784 TI - Interpolation of animal tracking data in a fluid environment. AB - Interpolation of geolocation or Argos tracking data is a necessity for habitat use analyses of marine vertebrates. In a fluid marine environment, characterized by curvilinear structures, linearly interpolated track data are not realistic. Based on these two facts, we interpolated tracking data from albatrosses, penguins, boobies, sea lions, fur seals and elephant seals using six mathematical algorithms. Given their popularity in mathematical computing, we chose Bezier, hermite and cubic splines, in addition to a commonly used linear algorithm to interpolate data. Performance of interpolation methods was compared with different temporal resolutions representative of the less-precise geolocation and the more-precise Argos tracking techniques. Parameters from interpolated sub sampled tracks were compared with those obtained from intact tracks. Average accuracy of the interpolated location was not affected by the interpolation method and was always within the precision of the tracking technique used. However, depending on the species tested, some curvilinear interpolation algorithms produced greater occurrences of more accurate locations, compared with the linear interpolation method. Total track lengths were consistently underestimated but were always more accurate using curvilinear interpolation than linear interpolation. Curvilinear algorithms are safe to use because accuracy, shape and length of the tracks are either not different or are slightly enhanced and because analyses always remain conservative. The choice of the curvilinear algorithm does not affect the resulting track dramatically so it should not preclude their use. We thus recommend using curvilinear interpolation techniques because of the more realistic fluid movements of animals. We also provide some guidelines for choosing an algorithm that is most likely to maximize track quality for different types of marine vertebrates. PMID- 16354785 TI - Hunting archer fish match their take-off speed to distance from the future point of catch. AB - Archer fish can shoot down insect prey with a sharp jet of water. Fish usually fire from positions that are not directly below their target so that a dislodged insect falls ballistically with a horizontal velocity component. Only 100 ms after the insect is on its path both the shooter and other school members can initiate a rapid turn and then head straight in the direction of the later point of impact of their falling prey. The quick turn and subsequent take-off are performed ;open-loop', based on the initial values of the falling insect's motion. We report here that archer fish can not only take off in the direction of the later point of impact but also predict its distance. Distance information allows the fish to adjust their take-off speed so that they would arrive within a narrow time slot slightly (about 50 ms) after their prey's impact, despite large differences in the size of the aligning turn and in the distance to be covered. Selecting a constant speed program with matched speed and catching the insect on the move minimizes frictional losses. The initial speed of starting fish is slightly but systematically too slow and is increased later so that the fish arrive 20 ms earlier than expected and often make the catch on a higher than take off speed. The variability of later speed changes suggests a systematic ;error' in the take-off, as if the fish underestimated distance. However, this apparent deficiency seems well adapted to the fish catching their prey at a high speed: if later the fish had no possibility to correct an initial error then it is better to start slightly too slow in order to minimize the risk of overshooting the point of catch. PMID- 16354786 TI - The absence of spatial echo suppression in the echolocating bats Megaderma lyra and Phyllostomus discolor. AB - Acoustic orientation most often takes place in echoic environments. The mammalian auditory system shows a variety of specializations to suppress misleading spatial information mediated by echoes. Psychophysically these specializations are summarized as the precedence effect. This study investigates how echolocating bats deal with multiple reflections of their sonar emissions from different spatial positions. In a two-alternative, forced choice paradigm, a study was made of the extent to which the echolocating bats Megaderma lyra and Phyllostomus discolor spontaneously suppress the spatial information of a second reflection of their sonar emission. The delay between the first and the second reflection ranged between 0 and 12.8 ms. In general, M. lyra (five individuals) and P. discolor (two individuals) did not suppress the spatial information of the second reflection of their sonar emission, whatever the delay. Only one M. lyra showed significant suppression for delays between 0.8 and 3.2 ms. However, this suppression could not be confirmed in an exact repetition of the experiment. The current data indicate that although bats may be able to suppress the spatial information of a second reflection, this is not their default mode of auditory processing. The reason for this exceptional absence of spatial echo suppression may lie in the shorter time constants of cochlear processing in the ultrasonic frequency range and the strong influence of cognitive components associated with the ;precedence effect'. PMID- 16354787 TI - Fixed metabolic costs for highly variable rates of protein synthesis in sea urchin embryos and larvae. AB - Defining the physiological mechanisms that set metabolic rates and the 'cost of living' is important for understanding the energy costs of development. Embryos and larvae of the sea urchin Lytechinus pictus (Verrill) were used to test hypotheses regarding differential costs of protein synthesis in animals differing in size, rates of protein synthesis, and physiological feeding states. For embryos, the rate of protein synthesis was 0.22+/-0.014 ng protein embryo(-1) h( 1) (mean +/- s.e.m.) and decreased in unfed larvae to an average rate of 0.05+/ 0.001 ng protein larva(-1) h(-1). Fed larvae had rates of synthesis that were up to 194 times faster than unfed larvae (9.7+/-0.81 ng protein larva(-1) h(-1)). There was no significant difference, however, in the cost of protein synthesis between these larvae with very different physiological states. Furthermore, the cost of synthesis in the larval stages was also similar to costs measured for blastula and gastrula embryos of 8.4+/-0.99 J mg(-1) protein synthesized. The cost of protein synthesis was obtained using both direct ('inhibitor') and indirect ('correlative') measurements; both methods gave essentially identical results. Protein synthesis accounted for up to 54+/-8% of metabolic rate in embryos. Percent of metabolism accounted for by protein synthesis in larvae was dependent on their physiological feeding state, with protein synthesis accounting for 16+/-4% in unfed larvae and 75+/-11% in fed larvae. This regulation of metabolic rate was due to differential rates of synthesis for a fixed energy cost per unit mass of protein synthesized. The cost of synthesizing a unit of protein did not change with increasing rates of protein synthesis. We conclude that the cost of protein synthesis is independent of the rate of synthesis, developmental stage, size and physiological feeding state during sea urchin development. PMID- 16354788 TI - Running over rough terrain: guinea fowl maintain dynamic stability despite a large unexpected change in substrate height. AB - In the natural world, animals must routinely negotiate varied and unpredictable terrain. Yet, we know little about the locomotor strategies used by animals to accomplish this while maintaining dynamic stability. In this paper, we perturb the running of guinea fowl with an unexpected drop in substrate height (DeltaH). The drop is camouflaged to remove any visual cue about the upcoming change in terrain that would allow an anticipatory response. To maintain stability upon a sudden drop in substrate height and prevent a fall, the bird must compensate by dissipating energy or converting it to another form. The aim of this paper is to investigate the control strategies used by birds in this task. In particular, we assess the extent to which guinea fowl maintain body weight support and conservative spring-like body dynamics in the perturbed step. This will yield insight into how animals integrate mechanics and control to maintain dynamic stability in the face of real-world perturbations. Our results show that, despite altered body dynamics and a great deal of variability in the response, guinea fowl are quite successful in maintaining dynamic stability, as they stumbled only once (without falling) in the 19 unexpected perturbations. In contrast, when the birds could see the upcoming drop in terrain, they stumbled in 4 of 20 trials (20%, falling twice), and came to a complete stop in an additional 6 cases (30%). The bird's response to the unexpected perturbation fell into three general categories: (1) conversion of vertical energy (EV=EP+EKv) to horizontal kinetic energy (EKh), (2) absorption of EV through negative muscular work (-DeltaEcom), or (3) converting EP to vertical kinetic energy (EKv), effectively continuing the ballistic path of the animal's center of mass (COM) from the prior aerial phase. However, the mechanics that distinguish these categories actually occur along a continuum with varying degrees of body weight support and actuation by the limb, related to the magnitude and direction of the ground reaction force (GRF) impulse, respectively. In most cases, the muscles of the limb either produced or absorbed energy during the response, as indicated by net changes in COM energy (Ecom). The limb likely begins stance in a more retracted, extended position due to the 26 ms delay in ground contact relative to that anticipated by the bird. This could explain the diminished decelerating force during the first half of stance and the exchange between EP and EK during stance as the body vaults over the limb. The varying degree of weight support and energy absorption in the perturbed step suggests that variation in the initial limb configuration leads to different intrinsic dynamics and reflex action. Future investigation into the limb and muscle mechanics underlying these responses could yield further insight into the control mechanisms that allow such robust dynamic stability of running in the face of large, unexpected perturbations. PMID- 16354789 TI - Molecular cloning and mRNA expression analysis of carp embryonic, slow and cardiac myosin heavy chain isoforms. AB - Three embryonic class II myosin heavy chains (MYHs) were cloned from the common carp (Cyprinus carpio L.), MYHemb1, MYHemb2 and MYHemb3. MYH DNA clones were also isolated from the slow muscle of adult carp acclimated to 10 degrees C (MYHS10) and 30 degrees C (MYHS30). Phylogenetic analysis demonstrated that MYHemb1 and MYHemb2 belonged to the fast skeletal muscle MYH clade. By contrast, the sequence of MYHemb3 was similar to the adult slow muscle isoforms, MYHS10 and MYHS30. MYHemb1 and MYHemb2 transcripts were first detected by northern blot analysis in embryos 61 h post-fertilization (h.p.f.) at the heartbeat stage, with peak expression occurring in 1-month-old juveniles. MYHemb1 continued to be expressed at low levels in 7-month-old juveniles when MYHemb2 was not detectable. MYHemb3 transcripts appeared at almost the same stage as MYHemb1 transcripts did (61 h.p.f.), and these genes showed a similar pattern of expression. Whole mount in situ hybridization analysis revealed that the transcripts of MYHemb1 and MYHemb2 were expressed in the inner part of myotome, whereas MYHemb3 was expressed in the superficial compartment. MYHS10 and MYHS30 mRNAs were first detected at hatching. In adult stages, the expression of slow muscle MYH mRNAs was dependent on acclimation temperature. MYHS10 mRNA was expressed at an acclimation temperature of 10 and 20 degrees C, but not at 30 degrees C. In contrast, MYHS30 mRNA was strongly expressed at all acclimation temperatures. The predominant MYH transcripts found in adult slow muscle and in embryos at hatching were expressed in adult fast muscle at some acclimation temperatures but not others. A MYH DNA clone was isolated from the cardiac muscle of 10 degrees C-acclimated adult fish (MYHcard). MYHcard mRNA was first detected at 61 h.p.f., but strong signals were only observed in the adult myocardium. The present study has therefore revealed a complex pattern of expression of MYH genes in relation to developmental stage, muscle type and acclimation temperature. None of the skeletal muscle MYHs identified so far was strongly expressed during the late juvenile stage, indicating further developmentally regulated members of the MYH II gene family remain to be discovered. PMID- 16354790 TI - Effect of subchronic treatment of memantine, galantamine, and nicotine in the brain of Tg2576 (APPswe) transgenic mice. AB - An increasing number of studies suggest that the present clinical therapy used in Alzheimer's disease (AD), in addition to having a symptomatic effect, also may interact with the ongoing neuropathological processes in the brain. The aim of this study was to investigate the effect of the cholinesterase inhibitor galantamine and the N-methyl-d-aspartate (NMDA) antagonist memantine in comparison to nicotine on the neuropathology of Tg2576 transgenic mice (APPswe). Nontransgenic and APPswe mice at 10 months of age were treated subcutaneously with saline, memantine, galantamine, or nicotine for 10 days. Nicotine reduced the guanidinium-soluble amyloid-beta peptide (Abeta) levels by 46 to 66%, whereas the intracellular Abeta levels remained unchanged. Treatment with nicotine also resulted in less glial fibrillary acidic protein immunoreactive astrocytes around the plaques, increased levels of synaptophysin, and increased number of alpha7 nicotinic acetylcholine receptors (nAChRs) in the cortex of APPswe transgenic mice. Galantamine treatment caused an increase in the cortical levels of synaptophysin in the APPswe mice. Memantine treatment reduced the total cortical levels of membrane-bound amyloid precursor protein (45-55%) in both transgenic and nontransgenic mice, which eventually may decrease the level of Abeta. In conclusion, galantamine, memantine, and nicotine have different interactions with Abeta processes, alpha7 nAChRs, and NMDA receptors in APPswe mice. These different effects might have therapeutic relevance, and this knowledge might be applicable to the development of new effective therapeutic strategies for AD. PMID- 16354791 TI - Talniflumate increases survival in a cystic fibrosis mouse model of distal intestinal obstructive syndrome. AB - Intestinal disease in cystic fibrosis (CF) mice closely mirrors aspects of obstructive syndromes in CF patients. The pathogenesis involves accumulation of mucoid debris in the crypts that fuse with intestinal content to form obstructing mucofeculant impactions. Treatment involves modalities that increase the fluidity of the luminal content, such as osmotic laxatives and liquid diets. We investigated the effects of talniflumate (Lomucin, Genaera Corporation, Plymouth Meeting, PA), a compound that may be beneficial to treatment of CF intestinal disease based on three mechanisms of action: mucus synthesis inhibition by blockade of the murine calcium-activated chloride channel 3 (mCLCA3), nonsteroidal anti-inflammatory effects, and inhibition of Cl(-)/HCO (-)(3) exchanger(s) involved in intestinal NaCl absorption. Cohorts of CF mice were fed control diet or diets containing either talniflumate (0.4 mg/g chow) or ibuprofen (0.4 mg/g chow) for 21 days to assess survival. Talniflumate significantly increased CF mouse survival from 26 to 77%, whereas ibuprofen had no effect (22% survival). Oral talniflumate did not alter crypt goblet cell numbers or change intestinal expression of mCLCA3 but tended to decrease crypt mucoid impaction. Ussing chamber studies indicated that talniflumate slightly increased the basal short-circuit current of CF intestine, but the change was not sensitive to secretagogue stimulation or bumetanide inhibition. In contrast, intracellular pH measurements of intact intestinal villous epithelium indicated that talniflumate significantly inhibited apical membrane Cl(-)/HCO (-)(3) exchange by >50%. We conclude that oral talniflumate increases the survival of CF mice, possibly by the beneficial effects of decreasing small intestinal NaCl absorption through the inhibition of apical membrane Cl(-)/HCO (-)(3) exchanger(s). PMID- 16354792 TI - The effects of deletions of the mouse Y chromosome long arm on sperm function- intracytoplasmic sperm injection (ICSI)-based analysis. AB - In mouse and man, Y chromosome deletions are frequently associated with spermatogenic defects. XY(Tdy)(m1)qdelSry males have an extensive Yq deletion that almost completely abolishes the expression of two gene families, Ssty and Sly, located within the male-specific region of the mouse Y long arm. These males exhibit severe sperm defects and sterility. XY(RIII)qdel males have a smaller interstitial Yq deletion, removing approximately two thirds of Ssty/Sly gene copies, and display an increased incidence of mild sperm head anomalies with impairment of fertility and an intriguing distortion in the sex ratio of offspring in favor of females. Here we used intracytoplasmic sperm injection (ICSI) to investigate the functional capacity of sperm from these Yq deletion males. Any selection related to the ability of sperm to fertilize in vitro is removed by ICSI, and we obtained two generations of live offspring from the infertile males. Genotyping of ICSI-derived offspring revealed that the Y(Tdym1)qdel deletion does not interfere with production of Y chromosome-bearing gametes, as judged from the frequency of Y chromosome transmission to the offspring. ICSI results for XY(RIII)qdel males also indicate that there is no deficiency of Y sperm production in this genotype, although the data show an excess of females following in vitro fertilization and natural mating. Our findings suggest that 1) Yq deletions in mice do not bias the primary sex ratio and 2) Y(RIII)qdel spermatozoa have poorer fertilizing ability than their X bearing counterparts. Thus, a normal complement of the Ssty and/or Sly gene families on mouse Yq appears necessary for normal sperm function. SUMMARY: ICSI was successfully used to reproduce infertile mice with Yq deletions, and the analysis of sperm function in obtained offspring demonstrated that gene families located within the deletion interval are necessary for normal sperm function. PMID- 16354793 TI - UTP14c is a recently acquired retrogene associated with spermatogenesis and fertility in man. AB - In the mouse, Utp14b is a retrogene transposed to an intron of Acsl3 (long-chain fatty-acid coenzyme A ligase 3) on mouse chromosome 1. It represents a copy of Utp14a, a ubiquitously expressed, X-linked gene involved in 18S rRNA synthesis. The Utp14b is specifically expressed in male germ cells and, when mutated in the jsd (juvenile spermatogonial depletion) mouse, results in early spermatogenic arrest and male infertility. To understand the function and relevance of the orthologous human gene in testis pathology, we mapped transcripts and searched for mutations within the gene in infertile males. In humans, the strict ortholog of UTP14b has degenerated and is no longer functional. However, a second active retroposon, UTP14c, is found within a widely expressed, putative glycosyl transferase-containing gene, GT8, on human chromosome 13. Unlike mouse Utp14b, which is only expressed in the male germ line, human UTP14c is expressed in testis and ovary, which is consistent with having a gonad-specific function. To determine if UTP14c is functionally equivalent to mouse Utp14b and essential to spermatogenesis in humans, we screened DNA from 234 nonobstructive, azoospermic/severely oligospermic males and 208 proven-fertile controls for mutations within UTP14c. We identified a mutation in three unrelated patients that introduces an in-frame stop codon truncating the UTP14c protein near the carboxyl terminus. These data indicate that UTP14c may be functionally equivalent to mouse Utp14b and required for normal male fertility in humans. The novel evolution of retroposed UTP14 genes supports the hypothesis that retrogenes play an important role in evolution via regulation of male reproductive fitness. PMID- 16354794 TI - Oleoylethanolamide protects human sperm cells from oxidation stress: studies on cases of idiopathic infertility. AB - N-acylethanolamides are naturally occurring hydrophobic molecules usually present in a very small amount in many mammalian tissues and cells. The presence of N acylethanolamides has also been demonstrated in human reproductive tracts and fluids, although their biological effects and molecular mechanisms of action are not yet completely elucidated. It is known that some N-acylethanolamides, such as oleoylethanolamide, have antioxidative properties. The aim of this study was to test whether oleoylethanolamide could protect sperm cells from reactive oxygen species-induced oxidative damage in cases of idiopathic infertility, because the excessive generation of these radicals was associated with this pathology. Our results show that 2.5 nM oleoylethanolamide in vitro supplementation significantly reduces DNA strand breaks both in fertile and infertile subjects. Moreover, oleoylethanolamide increases kinematic parameters, such as curvilinear velocity and amplitude of lateral head displacement and hyperactivation, both in the presence and in the absence of oxidative stress. Results of this study support the hypothesis of a possible protective action of oleoylethanolamide against reactive oxygen species, which could explain its beneficial effects on in vitro capacitated spermatozoa. PMID- 16354795 TI - The mouse T complex gene Tsga2, encoding polypeptides located in the sperm tail and anterior acrosome, maps to a locus associated with sperm motility and sperm egg interaction abnormalities. AB - Previous studies of sperm from mice heterozygous for a t haplotype (t) and heterospecific combinations of the t complex identified two tightly linked genetic factors responsible for t/t male sterility related to expression of the flagellar waveform aberration, curlicue. Dnahc8, an axonemal dynein heavy chain gene, is a strong candidate for the proximal factor, Ccua, but the identity of the distal factor, Ccub, is unknown. In the present study, we employ motility assays of sperm from males heterozygous for t and novel heterospecific combinations of the t complex to demonstrate that Ccub is a composite of at least two synergic elements, Ccub1, positioned within a genomic interval spanning approximately 0.6 Mb immediately distal to Dnahc8, and Ccub2, situated in a region approximately 4-7 Mb distal to Ccub1. We also show that Tsga2, a testis restricted gene, fulfills many of the prerequisites required to make it a strong candidate for Ccub1. These include: 1) its location within the aforementioned genomic interval; 2) a highly reduced level of testis expression by its heterospecific allele relative to the level of expression of its t allele; 3) determination that TSGA2(t) carries numerous nonsynonymous mutations in residues otherwise highly conserved in all known orthologous proteins; 4) the detection of major TSGA2 polypeptides in sperm protein extracts; and 5) the apparent distribution of these polypeptides in major sperm tail structures. Surprisingly, these TSGA2 isoforms appear to localize in the vicinity of the anterior acrosome, as well, suggesting that Tsga2 may also play a role in sperm-egg interaction. Finally, our results indicate that a TSGA2 polypeptide with apparent similarities to the smaller of the two sperm isoforms is expressed by epididymal cells. PMID- 16354796 TI - Clearance mechanisms of the aperistaltic oesophagus: the "pump gun" hypothesis. PMID- 16354797 TI - Concordance, disease progression, and heritability of coeliac disease in Italian twins. AB - BACKGROUND AND AIMS: We adopted the twin method to disentangle the genetic and environmental components of susceptibility to coeliac disease (CD). We estimated disease concordance rate by zygosity and HLA genotypes, discordance times, progression rates to disease, and heritability. METHODS: We crosslinked the Italian Twin Registry with the membership lists of the Italian Coeliac Disease Association and recruited 23 monozygotic (MZ) and 50 dizygotic (DZ) twin pairs with at least one affected member. Zygosity was assigned by DNA fingerprinting, and HLA-DQ and DR alleles were genotyped. Disease status was ascertained by antiendomysial, anti-human tissue transglutaminase antibodies, and bowel biopsy. RESULTS: Concordance was significantly higher in MZ (83.3% probandwise, 71.4% pairwise) than in DZ (16.7% probandwise, 9.1% pairwise) pairs. Concordance was not affected by sex or HLA genotype of the co-twin and being MZ was significantly associated with the occurrence of CD (Cox adjusted hazard ratio 14.3 (95% confidence interval 4.0-50.3)). In 90% of concordant pairs the discordance time was 3) samples from the phenotypically affected and non-penetrant subjects with R122H genotypes using allele specific quantitative reverse transcription-polymerase chain reaction (RT-PCR) and intron spanning nested RT-PCR followed by cDNA sequencing. Compared with PRSS1 mRNA levels, SPINK1 mRNA levels were low in normal appearing tissue but markedly increased in samples with chronic inflammation, independent of PRSS1 genotype. CONCLUSION: Attacks of acute pancreatitis in HP subjects appear to be independent of the relative expression of the mutant PRSS1 H122 allele or SPINK1 gene expression. The marked increase in SPINK1 gene expression with inflammation is consistent with its regulation as an acute phase protein. PMID- 16354800 TI - Natural course of lymphatic filariasis: insights from epidemiology, experimental human infections, and clinical observations. AB - Lymphatic filariasis has been described as a "spectral disease". Analysis of the natural course of infection in nonendemic individuals as well as experimental infections of "volunteers" suggests that the filarial parasites are not inherently aggressive infectious agents. Experimental infections of humans with infective larvae result in transient, low-level microfilaremia, if at all. Nonendemic individuals with limited exposure show no evidence of persistent infection or pathology. Nonendemic individuals exposed to repeated infections show accelerated pathology. It is tempting to speculate that normal, immunocompetent residents in an endemic area show either (a) no pathology (endemic normals) because they are subject to the relatively low levels of infection or (b) chronic pathology if they are repeatedly infected. It would appear that only those individuals rendered immunologically tolerant to filarial parasites become productively infected with the filarial parasites. The intensity of transmission may underlie the differences in clinical presentation seen in diverse global pockets of endemicity. PMID- 16354801 TI - Rapid identification of Leishmania complexes by a real-time PCR assay. AB - A real-time PCR assay for the detection of four Leishmania complexes (L. Viannia, L. mexicana, L. donovani/infantum, and L. major) was developed and evaluated. The assay was developed to detect the glucosephosphate isomerase gene and capitalizes on DNA sequence variability within that gene for Leishmania complex identification. Primer/probe sets were created and tested against a panel of 21 known negative controls and on DNA extracted from cultured promastigotes or from tissue biopsies from patients with cutaneous leishmaniasis. The assay was highly specific, as no amplification products were detected in the negative control samples while simultaneously retaining a high degree of complex-specific diagnostic accuracy for cultured organisms and patient clinical samples. Real time PCR offers rapid (within hours) identification of Leishmania to the complex level and provides a useful molecular tool to assist both epidemiologists and clinicians. PMID- 16354802 TI - A phase II dose-ranging study of sitamaquine for the treatment of visceral leishmaniasis in India. AB - This randomized, open label, multicenter study assessed the dose-response and safety profile for oral sitamaquine in 120 Indian subjects with visceral leishmaniasis (VL). Patients aged 5-64 years (mean age 21.2 years) received one of four sitamaquine doses (1.5, 1.75, 2.0, or 2.5 mg kg(-1) day(-1)) daily for 28 days. At Day 180 in the intent-to-treat population, final cure (primary efficacy outcome) was achieved in 92 of 106 (87%) patients overall and 25 of 31 (81%), 24 of 27 (89%), 23 of 23 (100%), and 20 of 25 (80%) patients at doses of 1.5, 1.75, 2.0, or 2.5 mg kg(-1) day(-1) sitamaquine, respectively. Sitamaquine was generally well tolerated. The most common adverse events during the active treatment phase were vomiting (8% [10 of 120]), dyspepsia (8% [9 of 120]) and cyanosis (3% [4 of 120]). Nephrotic syndrome (3% [3 of 120]) and glomerulonephritis (2% [2 of 120]) were also reported and require further investigation. Oral sitamaquine demonstrated efficacy in Indian VL and was well tolerated. PMID- 16354803 TI - Value of culture and nested polymerase chain reaction of blood in the prediction of relapses in patients co-infected with leishmania and human immunodeficiency virus. AB - The use of culture and a nested polymerase chain reaction (PCR) of blood in predicting the probability of relapse was evaluated in 20 patients co-infected with Leishmania and human immunodeficiency virus (HIV). Fourteen of 20 patients relapsed, with 24 clinical relapses diagnosed. During clinical relapse, the parasite was detected by culture in 21 of 24 blood samples and by nested PCR in 23 of 24 blood samples. After treatment and during asymptomatic periods, the parasite was detected by culture in 18 (19.1%) of 94 blood samples and by nested PCR in 58 (61.7%) of 94 blood samples. For positive blood cultures, the Kaplan Meier probability estimates for relapse at 6, 12, 18, and 24 months were 44%, 68%, 76%, and 76%, respectively, while for positive nested PCRs, the estimates were 20%, 33%, 45%, and 50%, respectively. For negative blood cultures, relapse probabilities for the same time points were 7%, 12%, 12%, and 12%, while for negative nested PCRs, these probabilities were 8%, 14%, 21%, and 26%. Nested PCR positive results in asymptomatic periods indicated presence of the parasite, but not necessarily relapse. However, the presence of viable parasites during post treatment follow-up increased the probability of relapse and showed that culture positivity could be a good relapse marker. PMID- 16354804 TI - Molecular diagnosis and typing of Trypanosoma cruzi populations and lineages in cerebral Chagas disease in a patient with AIDS. AB - Trypanosoma cruzi DNA was amplified from an intracranial biopsy and peripheral blood of an HIV patient with encephalitis; this episode was indicative of AIDS and congenital Chagas disease. The analysis of a micro-satellite locus revealed a multiclonal parasite population at the brain lesion with a more complex minicircle signature than that profiled in blood using restriction fragment length polymorphism (RFLP)-PCR and low stringency single primer (LSSP) PCR. Interestingly, different sublineages of T. cruzi II were detected in blood and brain by means of spliced-leader and 24salpha ribosomal-DNA amplifications. Quantitative-competitive PCR monitored the decrease of parasitic load during treatment and secondary prophylaxis with benznidazole. The synergy between parasiticidal plus anti-retroviral treatments probably allowed the patient a longer survival than usually achieved in similar episodes. This is the first case report demonstrating a differential distribution of natural parasite populations and sublineages in Chagas disease reactivation, showing the proliferation of cerebral variants not detectable in peripheral blood. PMID- 16354805 TI - Effect of Hurricane Isidore on Triatoma dimidiata distribution and Chagas disease transmission risk in the Yucatan Peninsula of Mexico. AB - Hurricanes can have devastating effects on health and may directly modulate vector-borne diseases. Chagas disease is a zoonosis caused by the protozoan parasite Trypanosoma cruzi and transmitted by triatomine bugs, and the effect of hurricanes on these bugs is largely unknown. We thus performed a detailed study of the changes in Triatoma dimidiata geographic distribution and infection rates after Hurricane Isidore devastated the Yucatan Peninsula in September 2002. Bugs were collected in 34 villages from the entire peninsula, during a year, starting 3 months after the hurricane. Pre- and posthurricane bug collections were compared to assess changes. The most notable effect was a large increase in domestic abundance of T. dimidiata during the 6 months after the hurricane. This increase was maximum along the path of the hurricane. These results suggest that vector control programs should be implemented along the path of hurricanes to prevent an increase in Chagas disease transmission risk in the ensuing months. PMID- 16354806 TI - Short report: persistent bradycardia caused by ciguatoxin poisoning after barracuda fish eggs ingestion in southern Taiwan. AB - We report an outbreak of ciguatoxin poisoning after barracuda fish ingestion in southern Taiwan. Three members of a family developed nausea, vomiting, watery diarrhea, and myalgias about 1 hour after eating three to ten eggs of a barracuda fish. Numbness of the lips and extremities followed the gastrointestinal symptoms about 2 hours after ingestion. Other manifestations included hyperthermia, hypotension, bradycardia, and hyperreflexia. Bradycardia persisted for several days, and one patient required a continuous infusion of intravenous atropine totaling 40 mg over 2 days. Further follow-up of the patients disclosed improvement of neurologic sequelae and bradycardia, but sensory abnormalities resolved several months later. In conclusion, ciguatoxin poisoning causes mainly gastrointestinal and neurologic effects of variable severity. In two patients with ciguatoxin poisoning after barracuda fish egg ingestion, persistent bradycardia required prolonged atropine infusion. PMID- 16354807 TI - Short report: Q fever and Plasmodium falciparum malaria co-infection in a patient returning from the Comoros archipelago. AB - Although Plasmodium falciparum malaria and Q fever are both prevalent in Africa, there have been no reports of co-infection to date. We report a case who returned from the Comoros archipelago diagnosed by serologic analysis as well as detection of Coxiella burnetii DNA in acute-phase serum. Thus, Q fever may be associated with malaria infection in travelers returning from disease-endemic countries. This diagnosis should be considered when the response to malaria treatment is incomplete. PMID- 16354808 TI - Avian hosts for West Nile virus in St. Tammany Parish, Louisiana, 2002. AB - West Nile virus (WNV) infections in free-ranging birds were studied in Slidell, St. Tammany Parish, Louisiana, after a human encephalitis outbreak peaked there in July 2002. Seroprevalence in resident, free-ranging wild birds in one suburban site was 25% and 24% in August and October, respectively, indicating that most transmission had ceased by early August. Mortality rates, seroprevalence rates, host competence, and crude population estimates were used in mathematical models to predict actual infection rates, population impacts, and importance as amplifying hosts for several common passerine birds. Northern cardinal (Cardinalis cardinalis) and house sparrow (Passer domesticus) were the principal amplifying hosts, but blue jay (Cyanocitta cristata) and northern mockingbird (Mimus polyglottos) also contributed. The blue jay population was reduced by an estimated 47%. A variety of passerine bird species combined to play an important role as amplifying hosts in the WNV transmission cycle. PMID- 16354809 TI - West Nile virus serosurveillance in Iowa white-tailed deer (1999-2003). AB - Sera from white-tailed deer (Odocoileus virginianus) were collected in Iowa during the winter months (1999-2003), 2 years before and after West Nile virus (WNV) was first reported in Iowa (2001), and were analyzed for antibodies to WNV. Samples from 1999 to 2001 were antibody negative by blocking enzyme-linked immunosorbent assay (bELISA) and plaque reduction neutralization test (PRNT(90)). Prevalence derived from bELISA (2002, 12.7%; 2003, 11.2%) and WNV PRNT(90) (2002, 7.9%; 2003, 8.5%) assays were similar. All samples were negative for antibodies against St. Louis encephalitis virus as determined by PRNT(90). Antibodies to flaviviruses were detected by indirect enzyme-linked immunosorbent assay (iELISA) prior to the first WNV cases reported in Iowa (1999-2001) with prevalence ranging from 2.2% to 3.2%, suggesting the circulation of an additional undescribed flavivirus prior to the introduction of WNV into the area. Flavivirus prevalence as determined by iELISA increased in 2002 and 2003 (23.3% and 31.9%, respectively). The increase in prevalence exceeded estimates of WNV prevalence, suggesting that conditions favored general flavivirus transmission (including WNV) during the 2002-2003 epizootic. These data indicate that serologic analysis of deer sera collected from hunter harvests may prove useful for surveillance and evidence of local transmission of WNV and other pathogens and identify white tailed deer as a species for further studies for host competency. PMID- 16354810 TI - A longitudinal study of Bayou virus, hosts, and habitat. AB - What is currently known about the ecology of North American hantaviruses has come largely from studies on Sin Nombre virus (SNV). We conducted a longitudinal study of Bayou virus (BAYV), the second-leading agent of hantavirus pulmonary syndrome in the United States. Antibodies to hantavirus were detected from Oryzomys palustris (most commonly infected species), Sigmodon hispidus, Peromyscus leucopus, Reithrodontomys fulvescens, and Baiomys taylori. However, only O. palustris had viral RNA in tissues and excreta, suggesting that antibodies detected in other species may have resulted from spill-over infection. Seroprevalence rates averaged around 16% for O. palustris and varied seasonally. The heaviest males exhibited the highest levels of seroprevalence. Seroprevalence was higher in coastal prairie (20.0%) than old-fields (10.5%) and was associated with host abundance. These patterns are similar to those of SNV and can be used in identification of potentially at-risk areas. PMID- 16354811 TI - Araguari virus, a new member of the family Orthomyxoviridae: serologic, ultrastructural, and molecular characterization. AB - This paper reports the results of serologic, structural, biochemical, and genetic studies indicating that Araguari virus, a previously unassigned viral agent, is a member of the family Orthomyxoviridae and genus Thogotovirus. Araguari virus has six RNA fragments; biologically, it shares several properties with other viruses in the family Orthomyxoviridae. Nucleotide sequencing of the RNA segments 4 (glycoprotein) and 5 (nucleoprotein) of Araguari virus aligned with the orthomyxoviruses, showing the closest relationship with Thogoto virus (sequence similarity = 61.9% and 69.1%, respectively, for glycoprotein and nucleoprotein), but also sharing a more distant similarity with Dhori and Influenza C viruses, especially for the glycoprotein gene. Based on these results, we propose that Araguari virus should be assigned as a new member of the family Orthomyxoviridae and genus Thogotovirus. PMID- 16354812 TI - Short report: assessment of the World Health Organization scheme for classification of dengue severity in Nicaragua. AB - The World Health Organization (WHO) scheme for classification of dengue severity was evaluated in a three-year study of 1,671 confirmed dengue cases in three Nicaraguan hospitals. The WHO classification of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) was compared with the presence of hemorrhagic manifestations, signs of vascular permeability, marked thrombocytopenia, and shock in 114 infants, 1,211 children, and 346 adults. We found that strict application of the WHO criteria fails to detect a significant number of patients with severe manifestations of dengue, especially in adults. PMID- 16354813 TI - Differences in dengue severity in infants, children, and adults in a 3-year hospital-based study in Nicaragua. AB - To investigate age-related differences in dengue severity, 114 infants, 1,211 children, and 346 adults with laboratory-confirmed dengue virus (DEN) infections presenting to three hospitals in major urban centers in Nicaragua were recruited from 1999 to 2001. The age distribution of dengue cases and the circulating serotype (predominantly DEN2) were representative of national data. Similar results were obtained when either dengue hemorrhagic fever/dengue shock syndrome or its principal manifestations (vascular permeability, internal hemorrhage, marked thrombocytopenia, and/or shock) were analyzed in relation to age and immune status. The burden of disease and of severe dengue was found predominantly in infants 4-9 months of age and in children 5-9 years old, and secondary DEN infection was a risk factor for severity in children. Age-related differences were identified in the prevalence of specific clinical manifestations as well as in their association with a confirmed DEN diagnosis. This represents one of the few comprehensive studies to analyze characteristics of dengue in infants, children, and adults in the same population and highlights age-related differences in dengue severity. PMID- 16354814 TI - Chromosomal differentiation of Anopheles funestus from Luanda and Huambo Provinces, western and central Angola. AB - The chromosomal polymorphism of Anopheles funestus sensu stricto from Angola was analyzed from indoor-resting samples collected in 11 peri-urban and rural sites of the Luanda and Huambo Provinces, which are > 450 km apart and have distinct eco-climatic conditions. Five polymorphic paracentric inversions were observed (scored chromatids range = 202 to 248): 2Ra, 2Rh, 3Ra, 3Rb, and 3La. Inversions 3Rb and 3La were highly polymorphic; the 2Ra and 3Ra arrangements were absent in Luanda. No significant departures from Hardy-Weinberg and linkage equilibria were found at the locality, commune, or province level (sites 50 chromosomes). Supervised discriminatory analyses were used to identify differentially expressed genes correlating with lineage and primary genetic change. The gene-expression profiles of normal hematopoietic cells were also studied. By using principal component analyses (PCA), a differentiation axis was exposed, reflecting lineages and maturation stages of normal hematopoietic cells. By applying the three principal components obtained from PCA of the normal cells on the leukemic samples, similarities between malignant and normal cell lineages and maturations were investigated. Apart from showing that leukemias segregate according to lineage and genetic subtype, we provide an extensive study of the genes correlating with primary genetic changes. We also investigated the expression pattern of these genes in normal hematopoietic cells of different lineages and maturations, identifying genes preferentially expressed by the leukemic cells, suggesting an ectopic activation of a large number of genes, likely to reflect regulatory networks of pathogenetic importance that also may provide attractive targets for future directed therapies. PMID- 16354840 TI - Microarray-based detection of genetic heterogeneity, antimicrobial resistance, and the viable but nonculturable state in human pathogenic Vibrio spp. AB - The morbidity and mortality associated with Vibrio-mediated waterborne diseases necessitates the development of sensitive detection technologies that are able to elucidate the identity, potential pathogenicity, susceptibility, and viability of contaminating bacteria in a timely manner. For this purpose, we have designed a single multiplex PCR assay to simultaneously amplify 95 diagnostic regions (encompassing species/serogroup-specific, antimicrobial resistance, and known toxin markers) and combined it with a long oligonucleotide microarray to create a platform capable of rapidly detecting and discriminating the major human pathogenic species from the genus Vibrio: V. cholerae, V. parahaemolyticus, V. vulnificus, and V. mimicus. We were able to validate this strategy by testing 100 geographically and temporally distributed isolates and observed an excellent concordance between species- and serotype-level microarray-based identification and traditional typing methods. In addition to accurate identification, the microarray simultaneously provided evidence of antibiotic resistance genes and mobile genetic elements, such as sulfamethoxazole-trimethoprim constins and class I integrons, and common toxin (ctxAB, rtxA, hap, hlyA, tl, tdh, trh, vvhA, vlly, and vmhA) and pathogenicity (tcpA, type III secretion system) genes that are associated with pathogenic Vibrio. The versatility of this method was further underscored by its ability to detect the expression of known toxin and virulence genes from potentially harmful viable but nonculturable organisms. The results suggest that this molecular identification method provides rapid and definitive information that would be of value in epidemiological, environmental, and health risk assessment surveillance. PMID- 16354841 TI - Make no bones about it: increasing epidemiologic evidence links vitamin D to pulmonary function and COPD. PMID- 16354842 TI - Rehabilitation and the national emphysema treatment trial. PMID- 16354843 TI - Health-care-associated pneumonia: a new therapeutic paradigm. PMID- 16354844 TI - A shift for the better. PMID- 16354845 TI - Pulmonary physicians in the practice of sleep medicine. PMID- 16354846 TI - Noninvasive ventilation for acute respiratory failure: but how severe? PMID- 16354847 TI - Relationship between serum 25-hydroxyvitamin d and pulmonary function in the third national health and nutrition examination survey. AB - CONTEXT: Age, gender, height, ethnicity, and smoking are important determinants of lung function but do not explain all of the variation between individuals. Low concentrations of vitamin D have been associated with a number of diseases, including osteoporosis, hypertension, and type I diabetes. It is possible that serum concentrations of vitamin D might also influence pulmonary function. OBJECTIVES: To determine the relationship between serum concentrations of 25 hydroxy vitamin D and pulmonary function. DESIGN, SETTING AND PARTICIPANTS: The analysis was conducted using data from the Third National Health and Nutrition Examination Survey, which was a cross-sectional survey of the US civilian population that was conducted from 1988 to 1994. The analyses were restricted to 14,091 people who > or = 20 years of age, were interviewed at mobile examination centers, and had undergone spirometry, and in whom serum 25-hydroxy vitamin D levels had been measured. RESULTS: After adjustment for age, gender, height, body mass index, ethnicity, and smoking history, the mean FEV1 was 126 mL (SE, 22 mL), and the mean FVC was 172 mL (SE, 26 mL) greater for the highest quintile of serum 25-hydroxy vitamin D level (> or = 85.7 nmol/L) compared with the lowest quintile (< or = 40.4 nmol/L; p < 0.0001). With further adjustment for physical activity, the intake of vitamin D supplements, milk intake, and the level of serum antioxidants, the mean difference between the highest and lowest quintiles of 25 hydroxy vitamin D was 106 mL (SE, 24 mL) for FEV1, and 142 mL (SE, 29 mL) for FVC (p < 0.0001). CONCLUSIONS: There is a strong relationship between serum concentrations of 25-hydroxy vitamin D, FEV1, and FVC. Further studies are necessary to determine whether supplementation with vitamin D is of any benefit in patients with respiratory disease. PMID- 16354849 TI - A multidimensional grading system (BODE index) as predictor of hospitalization for COPD. AB - STUDY OBJECTIVES: We hypothesized that the BODE (body mass index, airflow obstruction, dyspnea, and exercise capacity) index would better predict hospitalization for COPD than FEV1 alone, and the purpose of this study was to test this hypothesis in a cohort of patients with COPD. DESIGN: Historical cohort study. SETTING: University-affiliated hospital. PATIENTS: One hundred twenty seven patients with COPD recruited from the outpatient clinic of a single institution were followed up for a mean period of 16.2 months. MEASUREMENTS: The BODE index was calculated for each patient using variables obtained within 4 weeks of enrollment. The main outcome measure was the number of hospital admissions for COPD during follow-up. We used the Poisson regression model to quantify and compare the relationship between FEV1 and BODE scores with the number of hospital admissions. RESULTS: During the follow-up period, 47% of patients required at least one hospital admission and 17% died. Using Poisson regression analysis, a significant effect of BODE score on the number of hospital admissions was found (incidence rate ratio, 1.20; 95% confidence interval [CI], 1.15 to 1.25; p < 0.001). In comparison, there was a significant but smaller effect of the FEV1 percentage of predicted on the number of hospital admissions (incidence rate ratio, 0.08; 95% CI, 0.04 to 0.16; p < 0.001). When categorizing the BODE scores into four quartiles, we found that the BODE index is also a better predictor of hospital admissions than the staging system of COPD as defined by the Global Initiative for Chronic Obstructive Lung Disease. The pseudo r2 using quartiles of the BODE index as the predictor was 0.16, as compared to 0.04 for stages of severity based on FEV1. CONCLUSIONS: The BODE staging system, which includes in addition to FEV1 other physiologic and clinical variables, helps to better predict hospitalization for COPD. PMID- 16354850 TI - Effects of omega-3 polyunsaturated fatty acids on inflammatory markers in COPD. AB - BACKGROUND: COPD, the fifth-leading cause of death worldwide, is characterized by chronic inflammation. However, no available agent can effectively cure this inflammation. A dietary supplement containing omega-3 polyunsaturated fatty acids (PUFAs) has anti-inflammatory effects. In this study, we hypothesized that nutritional support with omega-3 PUFA-rich diets may be useful for treating COPD, and we compared the clinical features and inflammatory mediator levels between the COPD patients who received an omega-3 PUFA-rich supplement and those who received a nonrich supplement. METHODS: Sixty-four COPD patients received 400 kilocalories per day of an omega-3 PUFA-rich supplement (n-3 group) or an omega-3 PUFA-nonrich supplement (n-6 group) for 2 years. We prospectively investigated the clinical features of these patients and measured the levels of inflammatory mediators. RESULTS: In 6-min walk testing, the dyspnea Borg scale and decrease of arterial oxygen saturation measured by pulse oximetry significantly improved in the n-3 group. Leukotriene B4 levels in serum and sputum and tumor necrosis factor-alpha and interleukin-8 levels in sputum decreased significantly in the n 3 group, while there was no significant change in the n-6 group. Two patients in the n-3 group and three patients in the n-6 group had mild diarrhea, and three patients in the n-3 group and three patients in the n-6 group had nausea; however, their symptoms were controllable and they improved with treatment. With multiple regression analysis, it was proved that the omega-3 PUFA-rich diet significantly contributed to the change in cytokine levels in this study. CONCLUSION: We suggest nutritional support with an omega-3 PUFA-rich diet as a safe and practical method for treating COPD. PMID- 16354851 TI - Pattern of variables describing desaturator COPD patients, as revealed by cluster analysis. AB - STUDY OBJECTIVES: The aims of this study were to define, by cluster analysis, a pattern of clinical variables that differentiate desaturator (D) from nondesaturator (ND) patients affected by COPD, and to identify daytime variables that are predictive of nocturnal desaturation. PATIENTS: Fifty-one random, consecutive COPD outpatients (20 women; mean [+/- SD] age, 69.6 +/- 4.0 years) with mild daytime hypoxemia (Pao(2), 60 to 70 mm Hg) were enrolled into the study. Obstructive sleep apnea syndrome patients were excluded. MEASUREMENTS AND RESULTS: Lung volumes, arterial blood gas levels, and mean pulmonary artery pressure (MPAP) were measured, and nocturnal desaturation was evaluated with nighttime polygraphy. With least squares simple linear regression, the percentage of total recording time was highly correlated with a total nocturnal recording time of arterial oxygen saturation of < 90 mm Hg (T90) and MPAP (R = 0.84; R2= 71.20%); T90 was also highly correlated with daytime Paco2 (R = 0.70; R2= 48.96%). Multiple regression showed that T90 was highly correlated with both MPAP and Paco2 (R2= 97.75%). Hierarchical cluster analysis conducted with these three variables showed that D and ND patients differed in both nocturnal and daytime variables. The mean T90 was 30 +/- 3.5% in 19.2% and 8%, respectively, of the D and ND groups. Moreover, two D subgroups differing in MPAP and two ND subgroups differing in Paco2 were identified. CONCLUSIONS: D patients may be identified by a pattern of T90, MPAP, and Paco2 values, rather than by T90 alone, with the latter two variables being predictors of nocturnal desaturation severity. PMID- 16354852 TI - Skeletal muscle adaptations to interval training in patients with advanced COPD. AB - STUDY OBJECTIVES: To investigate the response to interval exercise (IE) training by looking at changes in morphologic and biochemical characteristics of the vastus lateralis muscle, and to compare these changes to those incurred after constant-load exercise (CLE) training. DESIGN: Randomized, controlled, parallel, two-group study (IE vs CLE training). SETTING: Multidisciplinary, outpatient, hospital-based, pulmonary rehabilitation program. PATIENTS: Nineteen patients with stable advanced COPD (mean +/- SEM FEV1, 40 +/- 4% predicted). INTERVENTIONS: Patients (n = 10) assigned to IE training exercised at a mean intensity of 124 +/- 15% of baseline peak exercise capacity (peak work rate [Wpeak]) with 30-s work periods interspersed with 30-s rest periods for 45 min/d. Patients (n = 9) allocated to CLE training exercised at a mean intensity of 75 +/ 5% Wpeak for 30 min/d. Patients exercised 3 d/wk for 10 weeks. MEASUREMENTS AND RESULTS: Needle biopsies of the right vastus lateralis muscle were performed before and after rehabilitation. After IE training, the cross-sectional areas of type I and IIa fibers were significantly increased (type I before, 3,972 +/- 455 microm2; after, 4,934 +/- 467 microm2 [p = 0.004]; type IIa before, 3,695 +/- 372 microm2; after, 4,486 +/- 346 microm2 [p = 0.008]), whereas the capillary-to fiber ratio was significantly enlarged (from 1.13 +/- 0.08 to 1.24 +/- 0.07 [p = 0.013]). Citrate synthase activity increased (from 14.3 +/- 1.4 to 20.5 +/- 4.2 micromol/min/g), albeit not significantly (p = 0.097). There was also a significant improvement in Wpeak (by 19 +/- 5%; p = 0.04) and in lactate threshold (by 17 +/- 5%; p = 0.02). The magnitude of changes in all the above variables was not significantly different compared to that incurred after CLE training. During training sessions, however, ratings of dyspnea and leg discomfort, expressed as fraction of values achieved at baseline Wpeak, were significantly lower (p < 0.05) for IE training (73 +/- 9% and 60 +/- 8%, respectively) compared to CLE training (83 +/- 10% and 87 +/- 13%, respectively). CONCLUSIONS: High-intensity IE training is equally effective to moderately intense CLE training in inducing peripheral muscle adaptations; however, IE is associated with fewer training symptoms. PMID- 16354853 TI - Risk factors for asthma and asthma severity in nonurban children in Connecticut. AB - STUDY OBJECTIVE: To examine asthma diagnosis, asthma severity, and the presence of established asthma risk factors in children who reside in nonurban communities. DESIGN: A cross-sectional study was conducted of 19,076 children (6 months to 18 years of age) who lived in 146 nonurban communities in the greater Hartford, CT, region and who were enrolled in a disease-management program (Easy Breathing II; Michelle Cloutier, MD; Hartford, CT) designed to improve asthma diagnosis and treatment. RESULTS: The overall frequency of physician-confirmed asthma in children seeking health care was 18%. Asthma frequency was related to low socioeconomic status (SES), non-Caucasian ethnicity, male gender, age > or = 5 years, and exposure to tobacco smoke, dust, or cockroaches in the multivariate analysis. When controlling for SES, African-American children were 1.33 times more likely (95% confidence interval [CI], 1.15 to 1.53) and Hispanic children were 1.60 times as likely (95% CI, 1.38 to 1.85) as Caucasian children to have asthma. In contrast, asthma severity was related to dust exposure, a family history of asthma, non-Caucasian ethnicity, and age < or = 4 years in the multivariate analysis. African-American children (odds ratio, 1.31; 95% CI, 1.03 to 1.67) had more severe asthma diagnosed as compared to Caucasian children. Hispanic ethnicity was not associated with an increase in asthma severity. CONCLUSION: Risk factors for asthma in nonurban children are similar to risk factors in urban children. Ethnicity is a risk factor for asthma regardless of SES. Even in nonurban environments, African-American and Hispanic children have more asthma, and African-American children have more severe disease than their Caucasian counterparts. PMID- 16354854 TI - Epidemiology and outcomes of health-care-associated pneumonia: results from a large US database of culture-positive pneumonia. AB - CONTEXT: Traditionally, pneumonia developing in patients outside the hospital is categorized as community acquired, even if these patients have been receiving health care in an outpatient facility. Accumulating evidence suggests that health care-associated infections are distinct from those that are truly community acquired. OBJECTIVE: To characterize the microbiology and outcomes among patients with culture-positive community-acquired pneumonia (CAP), health-care-associated pneumonia (HCAP), hospital-acquired pneumonia (HAP), and ventilator-associated pneumonia (VAP). DESIGN AND SETTING: A retrospective cohort study based on a large US inpatient database. PATIENTS: A total of 4,543 patients with culture positive pneumonia admitted into 59 US hospitals between January 1, 2002, and December 31, 2003, and recorded in a large, multi-institutional database of US acute-care hospitals (Cardinal Health-Atlas Research Database; Cardinal Health Clinical Knowledge Services; Marlborough, MA). MAIN MEASURES: Culture data (respiratory and blood), in-hospital mortality, length of hospital stay (LOS), and billed hospital charges. RESULTS: Approximately one half of hospitalized patients with pneumonia had CAP, and > 20% had HCAP. Staphylococcus aureus was a major pathogen in all pneumonia types, with its occurrence markedly higher in the non-CAP groups than in the CAP group. Mortality rates associated with HCAP (19.8%) and HAP (18.8%) were comparable (p > 0.05), and both were significantly higher than that for CAP (10%, all p < 0.0001) and lower than that for VAP (29.3%, all p < 0.0001). Mean LOS varied significantly with pneumonia category (in order of ascending values: CAP, HCAP, HAP, and VAP; all p < 0.0001). Similarly, mean hospital charge varied significantly with pneumonia category (in order of ascending value: CAP, HCAP, HAP, and VAP; all p < 0.0001). CONCLUSIONS: The present analysis justified HCAP as a new category of pneumonia. S aureus was a major pathogen of all pneumonias with higher rates in non-CAP pneumonias. Compared with CAP, non-CAP was associated with more severe disease, higher mortality rate, greater LOS, and increased cost. PMID- 16354855 TI - Nontuberculous mycobacterial infection: CT scan findings, genotype, and treatment responsiveness. AB - STUDY OBJECTIVE: The purpose of this study was to compare the imaging findings of nontuberculous mycobacterial (NTM) infection in patients with normal and abnormal cystic fibrosis (CF) genotypes, and normal and abnormal alpha1-antitrypsin (AAT) phenotypes. DESIGN: A retrospective review of medical records and chest CT scans from 85 patients with microbiologically proven NTM infection was performed. All patients had undergone genotype analysis for CF mutations, and phenotypic evaluation for AAT status. Patients with homozygous CF or AAT were not included. Two independent observers assessed the patterns and distribution of disease, according to a standardized score sheet. In 52 patients, follow-up CT scans were obtained 1 to 46 months (mean duration, 8 months) following the initial CT scan. The CT scan findings on the follow-up scan were visually compared with those on the initial CT scan for progression or regression of abnormalities. Statistical analysis was performed to evaluate the relationship between the dominant CT scan pattern and CF/AAT status, and between CT scan pattern and radiologic change on follow-up. RESULTS: Fifteen patients (18%) were found to carry a single CF mutation, and an abnormal AAT phenotype was seen in 13 patients (15%). Three patients (3%) were found to have both a heterozygous CF mutation and a heterozygous AAT phenotype. On the initial CT scans, bronchiectasis and nodules were the most frequent findings of NTM infection in all three groups (p > 0.05). The prevalence of nodules was slightly lower in the CF group, and the prevalence of linear scarring was greater in the AAT group than in the normal group (p < 0.05). Among the 52 patients who had a follow-up CT scan, 8 (15%) had a CF mutation and 6 (12%) had an abnormal AAT phenotype. The extent and pattern of abnormality seen on the initial CT scan did not predict change on follow-up evaluation. After treatment, 40 patients (56%) with a normal CF genotype had decrease in disease extent, compared with 4 patients (25%) with a CF mutation (p < 0.05). Bronchiectasis was improved in approximately 35% of those with normal genotype, but in none of those with a CF mutation. CONCLUSION: In patients with NTM infection, the CT scan findings show only minor differences according to phenotype and genotype. Initial CT scan findings do not predict change on follow up CT scan evaluation. However, on follow-up CT scan, patients with CF mutations are less likely to improve, while those with AAT phenotype appear to have a radiographic outcome similar to those with normal phenotype. PMID- 16354856 TI - Relationship of dyspnea to respiratory drive and pulmonary function tests in obese patients before and after weight loss. AB - BACKGROUND: Dyspnea is a common complaint in obese patients, who also frequently have abnormal pulmonary function test (PFT) results without evidence of lung disease. We studied the relationship between dyspnea, PFT results, and respiratory drive in morbidly obese patients before and after weight loss. METHOD: Twenty-eight obese patients underwent PFTs including spirometry, lung volume measurements, and ventilatory drive assessment using the carbon dioxide rebreathing technique. The score of the dyspnea portion of the Chronic Respiratory Disease Questionnaire (CRQ) was used to assess dyspnea. CRQ and respiratory drive measurements were repeated in 10 patients after induced weight loss by gastroplasty RESULTS: Mean +/- SD body mass index (BMI) prior to surgery was 47 +/- 6.5 kg/m2. Patients were then classified into two groups: group 1, mild-to-moderate dyspnea (dyspnea score > 4); and group 2, severe dyspnea (dyspnea score < 4). Group 2 had higher respiratory drive parameters and significantly lower lung volumes compared to group 1. After gastroplasty, there were significant reductions in BMI (p = 0.000), dyspnea score (p = 0.000), occlusion pressure 100 ms after the start of inspiration (P100) at end-tidal carbon dioxide (ETCO2) of 60 mmHg (p = 0.011), minute ventilation (Ve) at ETCO2 of 60 mmHg, and Ve slope (0.017). P100 slope was reduced, but it did not reach statistical significance. CONCLUSION: The degree of dyspnea commonly observed in obese patients can be explained, in part, by increased ventilatory drive and reduced static lung volumes. Gastroplasty results in a significant reduction in BMI and respiratory drive measurements as well as significant improvement in dyspnea. PMID- 16354848 TI - The effects of pulmonary rehabilitation in the national emphysema treatment trial. AB - STUDY OBJECTIVES: Pulmonary rehabilitation is an established treatment in patients with chronic lung disease but is not widely utilized. Most trials have been conducted in single centers. The National Emphysema Treatment Trial (NETT) provided an opportunity to evaluate pulmonary rehabilitation in a large cohort of patients who were treated in centers throughout the United States. DESIGN: Prospective observational study of cohort prior to randomization in a multicenter clinical trial. SETTING: University-based clinical centers and community-based satellite pulmonary rehabilitation programs. PATIENTS AND INTERVENTION: A total of 1,218 patients with severe emphysema underwent pulmonary rehabilitation before and after randomization to lung volume reduction surgery (LVRS) or continued medical management. Rehabilitation was conducted at 17 NETT centers supplemented by 539 satellite centers. MEASUREMENTS AND RESULTS: Lung function, exercise tolerance, dyspnea, and quality of life were evaluated at regular intervals. Significant (p < 0.001) improvements were observed consistently in exercise (cycle ergometry, 3.1 W; 6-min walk test distance, 76 feet), dyspnea (University of California, San Diego Shortness of Breath Questionnaire score, -3.2; Borg breathlessness score: breathing cycle, -0.8; 6-min walk, -0.5) and quality of life (St. George Respiratory Questionnaire score, -3.5; Quality of Well-Being Scale score, +0.035; Medical Outcomes Study 36-item short form score: physical health summary, +1.3; mental health summary, + 2.0). Patients who had not undergone prior rehabilitation improved more than those who had. In multivariate models, only prior rehabilitation status predicted changes after rehabilitation. In 20% of patients, exercise level changed sufficiently after rehabilitation to alter the NETT subgroup predictive of outcome. Overall, changes after rehabilitation did not predict differential mortality or improvement in exercise (primary outcomes) by treatment group. CONCLUSIONS: The NETT experience demonstrates the effectiveness of pulmonary rehabilitation in patients with severe emphysema who were treated in a national cross-section of programs. Pulmonary rehabilitation plays an important role in preparing and selecting patients for surgical interventions such as LVRS. PMID- 16354857 TI - The effects of ethanol ingestion on the accuracy of pulmonary diffusing capacity measurement. AB - BACKGROUND: Erroneous diffusing capacity of the lung for carbon monoxide (Dlco) values as measured by spectrophotometry were observed at our facility in ethanol intoxicated subjects. An atypical methane curve tracing was noted in these subjects. STUDY OBJECTIVES: We hypothesized that ethanol intoxication interferes with Dlco measurements obtained using methane and designed a study to assess the blood ethanol level at which this occurs. STUDY DESIGN: Dlco and breath ethanol levels were measured at baseline and after escalating doses of ethanol in seven healthy subjects. SETTING: Pulmonary function testing laboratory of a university hospital. PARTICIPANTS: Seven healthy volunteers. INTERVENTIONS: Dlco measurement at baseline and after escalating doses of ethanol. MEASUREMENTS AND RESULTS: We found no significant change in measured Dlco values for a wide range of blood ethanol levels (from 0.006 to 0.12 mg/dL). However, subsequently, an abnormal methane curve and Dlco were again observed in an intoxicated subject whose ethanol blood level was 0.22 mg/dL. CONCLUSIONS: We conclude that interference between breath ethanol level and Dlco measurement exists only for blood ethanol levels well above the legal limit for intoxication. Based on our observations, we suggest that ethanol intoxication should be suspected when an abnormal Dlco measurement occurs along with an abnormal methane curve tracing. If intoxication is confirmed, precautions should be taken and referral for treatment of possible ethanol dependence should be offered. PMID- 16354858 TI - Maximal airway response to methacholine in cough-variant asthma: comparison with classic asthma and its relationship to peak expiratory flow variability. AB - BACKGROUND: In asthmatic subjects, not only airway sensitivity but maximal airway response are increased on the dose-response curve to methacholine, and peak expiratory flow (PEF) variability is closely related to airway hypersensitivity and maximal airway response. OBJECTIVE: The aims of this study were to compare the prevalence and the level of maximal response plateau between patients with cough-variant asthma (CVA) and those with classic asthma (CA), and to examine the relationship between airway hypersensitivity or maximal airway response and PEF variability in patients with CVA. METHODS: A high-dose methacholine inhalation test was performed on 83 patients with CVA and on 83 patients with CA matched for provocative concentration of methacholine causing a 20% fall in FEV1 (PC20). PEF was recorded in the morning and evening for 14 consecutive days in 78 CVA patients, and the amplitude percentage mean was used to express the diurnal PEF variation. RESULTS: Fifty-two CVA subjects (62.7%) but only 33 CA subjects (39.8%) showed a maximal response plateau. This difference was significant after correction by the Bonferroni method (corrected p = 0.024). Subjects in the CVA and CA groups showing a plateau had significantly different plateau levels (38.0 +/- 5.9% vs 42.9 +/- 3.9%, corrected p = 1.0 x 10(-4)). In patients with CVA, no significant relationship was found between PC20 and PEF variability. However, the absence of a maximal response plateau and a higher plateau level were associated with increased PEF variability. CONCLUSIONS: Maximal airway response may be an important confounder in the relationship between airway hypersensitivity and the clinical expression of asthma. The identification of a maximal response plateau and the level of this plateau in patients with CVA provide information relevant to PEF variability. PMID- 16354859 TI - Lipomatous hypertrophy of the interatrial septum: prevalence and features on fusion 18F fluorodeoxyglucose positron emission tomography/CT. AB - OBJECTIVE: To determine the prevalence of lipomatous hypertrophy of the interatrial septum (LHIS) on CT and its metabolic pattern on 18F fluorodeoxyglucose (FDG)-positron emission tomography (PET). METHOD AND MATERIALS: Eight hundred two CT PET scans were reviewed. Patients were included if the interatrial septum was > or = 1 cm and excluded if there was evidence of malignancy in the adjacent lung, hilum, or mediastinum. CT scans were fused with PET scans, and the mean standardized uptake value (SUV) was calculated over the LHIS, chest wall (CW) fat, and mediastinal blood pool. CT scans were reviewed for presence of excessive fat in the mediastinum, pericardial, peridiaphragmatic, peritoneal, and retroperitoneal regions and for the presence of emphysema. Medical records were reviewed for body mass index (BMI) and history of arrhythmia. RESULTS: Twenty-three of 802 patients (2.8%) had LHIS on CT (9 women and 14 men); average age was 75.6 years (range, 58 to 95 years). Average BMI of 17 patients (+/- SD) was 31 +/- 4.9 (range, 22.1 to 39.9). Mean CT values were as follows: thickening of LHIS, 1.47 +/- 0.35 cm (range, 1.07 to 2.25 cm); LHIS, - 79.6 + 24.5 Hounsfield unit (HU) [range, - 11 to - 121 HU]. LHIS was dumbbell shaped in 18 patients. Mean SUVs were as follows: LHIS, 1.84 +/- 0.10 (range, 0.48 to 3.48); CW fat, 0.36 + 0.37 (range, 0.04 to 1.98); blood pool, 1.74 + 0.51 (range, 0.25 to 2.71). The SUV of LHIS was greater than the SUV of CW wall fat in all patients (p < 0.0001). There was significant correlation between SUV and thickness of the LHIS on CT (p < 0.0001, r = 0.883). Those with dumbbell-shaped LHIS (p < 0.003) and presence of emphysema (p < 0.0377) had greater LHIS mean SUV. CONCLUSION: The SUV of LHIS was greater than the SUV of CW fat in all patients. LHIS with greater thickness or dumbbell shape had greater FDG uptake. These findings on CT and PET are important to recognize in order to avoid false positive FDG-PET interpretations. PMID- 16354860 TI - Contrast-enhanced sonography for differential diagnosis of pleurisy and focal pleural lesions of unknown cause. AB - BACKGROUND: Ultrasound enables the visualization of pleural-based lesions with a poor correlation to specific pathology. At this time, there are no data about the diagnostic value of contrast-enhanced sonography (CES) in pleural lesions. METHODS: From August 2004 to January 2005, 25 consecutive patients with clinical symptoms of pleurisy and focal pleural lesions of unknown origin seen on B-mode ultrasonography were prospectively studied by CES. The lesions were diagnosed as pleuropneumonia (n = 12), pulmonary embolism/infarction (n = 7), malignant lymphoma (n = 2), pleural metastasis (n = 2), granuloma (n = 1), and unknown cause (n = 1). The diagnosis of the lesions was confirmed by contrast-enhanced CT scanning (n = 20), scintigraphy (n = 3), and follow-up (n = 2). Time to the enhancement of the contrast agent was determined. The CES patterns were evaluated during the arterial phase (ie, 2 to 30 s) and the parenchymal phase (ie, 1 to 5 min). The extent of the enhancement of pleural lesions was classified using normal liver tissue as an in vivo reference (absent, hypoechoic, isoechoic, hyperrechoic, or mixed echogenicity). RESULTS: In 20 patients, an enhancement of the pleural lesion was seen. All 12 patients with pleuropneumonia had a short time to enhancement (between 1 and 6 s), and a marked enhancement (isoechoic/hyperechoic) during the arterial and parenchymal phase. In the remaining 13 patients with other diagnoses than pleuropneumonia, 5 patients had no enhancement and 8 patients had a delayed time to enhancement (> 6 s). The extent of the enhancement was reduced (hypoechoic/anechoic) in 12 of 13 patients during the arterial and parenchymal phases. CONCLUSION: In patients with pleurisy and pleural lesions of unknown cause that were found sonographically, CES enables the diagnosis or exclusion of pleuropneumonia. PMID- 16354861 TI - Frequency and correlates of adverse events in a respiratory diseases hospital in Mexico city. AB - OBJECTIVES: To estimate the frequency of severe adverse events (AEs) during hospital stay as well as their type and correlates in a referral hospital for respiratory diseases, using methods similar to those used in the Harvard Medical Malpractice Study. DESIGN: Retrospective review of medical records in a stratified sample of 836 patients drawn from a total of 4,555 hospital admissions registered during the year 2001. SETTING: A referral tertiary-care hospital for patients with respiratory diseases located in Mexico City. MAIN OUTCOME MEASURES: Weighted prevalence of AEs and odds ratios for correlates. RESULTS: The overall weighted prevalence for AEs was 9.1% (95% confidence interval, 7.5 to 10.4%). Of these patients, 17% had a related transient disability, 52% had a prolonged hospital stay, and 26% had an AE that according to the reviewers contributed to their death. Of the total number of AEs, 74% were qualified as potentially preventable. Among all types of AEs, we identified as most relevant for a chest hospital the delayed surgical treatment of empyema, representing 11% of the total. CONCLUSIONS: The frequency of AEs in a tertiary-care respiratory hospital is similar to that reported in general hospitals. A strategy to improve the treatment of empyema is needed. PMID- 16354862 TI - The incidence of tracheoarterial fistula in patients with chronic tracheostomy tubes: a retrospective study of 544 patients in a long-term care facility. AB - OBJECTIVE: The incidence of tracheoarterial fistula (TAF) in patients with short term tracheostomy tubes has been reported between 0.6% and 0.7%. The purpose of this study was to determine the incidence of TAF in patients with long-term tracheostomy tubes used for the management of chronic respiratory failure. SETTING: Long-term ventilator facility. DESIGN: Retrospective. METHODS: Medical records of 544 patients admitted to our institution between January 1981 and December 2002 were reviewed. All patients underwent tracheostomy prior to admission to our facility. Patient age, length of stay (LOS), duration of tracheostomy, and serum albumin levels were compared between patients with and without TAF; p values were obtained using the Student unpaired t test for equal variances. RESULTS: TAF was diagnosed in five patients. The incidence of TAF in our population was 0.7%. The average age of these patients was significantly less than the study population (31.20 years vs 68.27 years). When one patient outlier was eliminated, LOS was not significant. CONCLUSION: TAF is an uncommon complication of tracheostomy tubes. The incidence of TAF in patients with long term tracheostomy tubes is similar to that reported in short-term tracheostomy tubes. PMID- 16354863 TI - Introduction of a 14-hour work shift model for housestaff in the medical ICU. AB - STUDY OBJECTIVE: To describe the outcomes of switching housestaff from a traditional model of "long-call" every 4 days to a 14-h work-shift model in a medical ICU (MICU) over a 5-week pilot period. DESIGN: Retrospective comparison of a 5-week pilot period for a 14-h work-shift model vs a 4-month period for the traditional model. SETTING: The MICU of a tertiary medical center. PARTICIPANTS: A total of 626 patients admitted to the MICU and 34 internal medicine residents taking care of them. INTERVENTIONS: None. MEASUREMENTS: Severity-adjusted patient outcomes, housestaff performance on end-of-rotation examinations, and scheduled duty hours during the 5-week 14-h work-shift pilot period compared to a 16-week traditional nonpilot work period. RESULTS: There were no statistically significant differences in patients' adjusted mortality rates, hospital lengths of stay, or housestaff performance on end-of-rotation knowledge assessment examinations between the pilot and nonpilot periods. During the pilot period, each resident was scheduled to work for an average of 61.3 h weekly, and each fellow for 65.3 h weekly. In comparison, each resident and fellow was scheduled to work for an average of 73.3 h weekly during the nonpilot period. CONCLUSIONS: The 14-h work shift is a feasible option for housestaff rotation in the MICU. Although the power of our study to detect significant differences in mortality, length of stay, and educational outcomes was low, there was no evidence of compromised patient care or housestaff education associated with the 14-h shift model over the course of this 5-week pilot study. PMID- 16354864 TI - Noninvasive vs conventional mechanical ventilation in acute respiratory failure: a multicenter, randomized controlled trial. AB - STUDY OBJECTIVE: Noninvasive mechanical ventilation (NIMV) is beneficial for patients with acute respiratory failure (ARF) when added to medical treatment. However, its role as an alternative to conventional mechanical ventilation (CMV) remains controversial. Our aim was to compare the efficacy and resource consumption of NIMV against CMV in patients with ARF. DESIGN: A randomized, multicenter, controlled trial. SETTING: Seven multipurpose ICUs. PATIENTS: Sixty four patients with ARF from various causes who fulfilled criteria for mechanical ventilation. INTERVENTION: The noninvasive group received ventilation through a face mask in pressure-support mode plus positive end-expiratory pressure; the conventional group received ventilation through a tracheal tube. MEASUREMENTS AND RESULTS: Avoidance of intubation, mortality, and consumption of resources were the outcome variables. Thirty-one patients were assigned to the noninvasive group, and 33 were assigned to the conventional group. In the noninvasive group, 58% patients were intubated, vs 100% in the conventional group (relative risk reduction, 43%; p < 0.001). Stratification by type of ARF gave similar results. In the ICU, death occurred in 23% and 39% (p = 0.09) and complications occurred in 52% and 70% (p = 0.07) in the noninvasive and conventional groups, respectively. There were no differences in length of stay. The Therapeutic Intervention Score System-28, but not the direct nursing activity time, was lower in the noninvasive group during the first 3 days. CONCLUSIONS: NIMV reduces the need for intubation and therapeutic intervention in patients with ARF from different causes. There is a nonsignificant trend of reduction in ICUs and hospital mortality together with fewer complications during ICU stay. PMID- 16354865 TI - Impact of a disease management program upon caregivers of chronically critically ill patients. AB - OBJECTIVES: Few studies have examined the effects of caregiving on the caregivers of chronically critically ill (CCI) patients, and no one has examined the impact of a disease management program (DMP) on physical and psychological outcomes for the caregivers of CCI patients. The purposes of this study of caregivers of CCI patients were as follows: (1) to describe the characteristics of CCI patients and caregivers and to examine the frequency of depression, subjective burden, and physical health; (2) to examine factors related to depression after hospital discharge; and (3) to examine the effects of a DMP on the physical health, depression, and burden of caregivers 2 months post-hospital discharge. DESIGN: Prospective experimental design. SETTING AND PARTICIPANTS: Caregivers of 290 patients who had received > 3 days of mechanical ventilation while in the ICU of a university medical center. MEASUREMENTS: Sociodemographics, caregiver burden, physical health status, and depression were measured using established tools. RESULTS: Interviews of caregivers were conducted at hospital discharge and 2 months later. Seventy-three percent of patients survived, completed the study period, and required caregiving 2 months later. Caregivers of patients residing in an institution reported higher depression (p = 0.0001), higher burden (ie, disrupted schedule, p = 0.0001; lack of family support, p = 0.036), and greater health problem scores (p = 0.0001) than did caregivers of patients residing at home. The DMP did not have a statistically significant impact on any of the outcome variables. However, by 2 months, 54% of caregivers in the experimental group had no depression or mild depression compared with 34.5% of the control group. CONCLUSION: Two months after hospital discharge, approximately 25% of caregivers were classified as depressed with 16.7% of the depressed group classified as moderately or severely depressed. The caregivers of CCI patients are at risk for post-hospital discharge depression, and the caregivers of institutionalized CCI patients are at highest risk of long-term negative effects from caregiving. PMID- 16354866 TI - Management of patients requiring prolonged mechanical ventilation: report of a NAMDRC consensus conference. AB - Patients requiring prolonged mechanical ventilation (PMV) are rapidly increasing in number, as improved ICU care has resulted in many patients surviving acute respiratory failure only to then require prolonged mechanical ventilatory assistance during convalescence. This patient population has clearly different needs and resource consumption patterns than patients in acute ICUs, and specialized venues, management strategies, and reimbursement schemes for them are rapidly emerging. To address these issues in a comprehensive way, a conference on the epidemiology, care, and overall management of patients requiring PMV was held. The goal was to not only review existing practices but to also develop recommendations on a variety of assessment, management, and reimbursement issues associated with patients requiring PMV. Formal presentations were made on a variety of topics, and writing groups were formed to address three specific areas: epidemiology and outcomes, management and care settings, and reimbursement. Each group was charged with summarizing current data and practice along with formulation of recommendations. A working draft of the products of these three groups was then created and circulated among all participants. The document was reworked with input from all concerned until a final product with consensus recommendations on 12 specific issues was achieved. PMID- 16354867 TI - Bronchopleural fistulas: an overview of the problem with special focus on endoscopic management. AB - A bronchopleural fistula (BPF) is a communication between the pleural space and the bronchial tree. Although rare, BPFs represent a challenging management problem and are associated with high morbidity and mortality. By far, the postoperative complication of pulmonary resection is the most common cause, followed by lung necrosis complicating infection, persistent spontaneous pneumothorax, chemotherapy or radiotherapy (for lung cancer), and tuberculosis. The treatment of BPF includes various surgical and medical procedures, and of particular interest is the use of bronchoscopy and different glues, coils, and sealants. Localization of the fistula and size may indicate potential benefits of surgical vs endoscopic procedures. In high-risk surgical patients, endoscopic procedures may serve as a temporary bridge until the patient's clinical status is improved, while in other patients endoscopic procedures may be the only option. Therapeutic success has been variable, and the lack of consensus suggests that no optimal therapy is available; rather, the current therapeutic options seem to be complementary, and the treatment should be individualized. PMID- 16354868 TI - Exercise-induced bronchoconstriction in athletes. AB - Exercise-induced bronchoconstriction (EIB) describes airway narrowing that occurs in association with exercise. EIB occurs in up to 90% of asthmatic patients and is estimated to occur in > 10% of the general population. Recent reviews have identified asthma as a risk factor for sudden death and have reported many deaths that have been attributed directly to EIB. We present a review of the literature related to EIB in athletes including sections discussing its pathogenesis, diagnosis, and treatment, and which athletes are most at risk for experiencing EIB. PMID- 16354869 TI - Targeted therapy for the treatment of advanced non-small cell lung cancer: a review of the epidermal growth factor receptor antagonists. AB - Lung cancer is the most common cause of cancer death. The vast majority of patients present with non-small cell lung cancer (NSCLC) in advanced inoperable stages. The current first-line treatment for patients with advanced NSCLC includes chemotherapy and palliative radiotherapy, but most patients relapse and eventually succumb to the disease. Advances in our knowledge of cancer cell biology have led to the development of specific molecular-targeted therapeutic agents. Mutations in the epidermal growth factor receptor (EGFR) have been identified in NSCLC cells, and overexpression of the EGFR and its ligands is a common feature of many cancers; therefore, EGFR has become an attractive target for various antitumor strategies. Aberrant signaling from the EGFR is known to be important in the development and progression of NSCLC. Two oral EGFR inhibitors, gefitinib and erlotinib, are small-molecule agents that selectively inhibit the intracellular tyrosine kinase activity of the EGFR. Both have demonstrated antitumor activity in patients with advanced NSCLC who have failed all prior treatment regimens. In addition, the anti-EGFR monoclonal antibody cetuximab has shown promising activity in both first-line and second-line settings in patients with advanced NSCLC. Furthermore, patients with severe comorbidities who would not be eligible for systemic chemotherapy are candidates for these targeted therapies. Finally, these agents have also been shown to be effective for relieving symptoms, maintaining stable disease, and improving quality of life without the adverse events that may be associated with cytotoxic cancer therapies. This report will review the mechanism of action, indications, contraindications, patient selection, and efficacy and side effects of this new class of compounds. It is important for pulmonologists to be aware of this class of compounds, as they can provide benefit to patients with NSCLC who may not have been previously considered for antitumor therapy. PMID- 16354870 TI - Pitfalls in diagnosis and clinical, echocardiographic, and hemodynamic findings in endomyocardial fibrosis: a 25-year experience. AB - Endomyocardial fibrosis (EMF) is a fascinating disease entity of unknown etiology. It is prevalent in the tropical zone. Its essential features are the formation of fibrous tissue on the endocardium and to a lesser extent in the myocardium of the inflow tract and apex of one or both ventricles. It results in endocardial rigidity, atrioventricular valve incompetence secondary to papillary muscle involvement, and progressive reduction of the cavity of the involved ventricle leading to restriction in filling and atrial enlargement. This article will present 21 patients with EMF who were initially referred to our hospital from 1979 to 2004 with different diagnoses: rheumatic heart disease with mitral and or tricuspid regurgitation (n = 9), constrictive pericarditis (n = 6), restrictive cardiomyopathy (n = 1), hypertrophic cardiomyopathy apical type (n = 2), dilated cardiomyopathy (n = 2), and Ebstein malfunction of the tricuspid valve (n = 1). The clinical, echocardiographic, hemodynamic, and angiographic findings in these 21 patients are presented; echocardiographic findings lead to the right diagnosis. The presence of a small ventricle with obliteration of the apex and large atrium shown on two-dimensional echocardiography is highly suggestive of EMF. PMID- 16354871 TI - A bibliometric analysis of global research production in respiratory medicine. AB - STUDY OBJECTIVES: To evaluate the contribution of different world regions in respiratory research productivity. METHODS: The world was divided into nine regions based on a combination of geographic, economic, and scientific criteria. Using the PubMed database, we retrieved information about the origin of articles from 30 journals included in the Respiratory System category of the Journal Citation Reports database for a 9-year period (1995 to 2003). We estimated the total number of publications, their mean impact factor, the product of these two parameters, and the research productivity per million of population of the world area divided by the gross national income per capita (GNIPC), for every year and the whole period of the study, for all defined world regions. MEASUREMENTS AND RESULTS: Data on the country of origin of the publications was available for 48,614 of 49,382 retrieved articles (98.5%). The majority of articles published between 1995 and 2003 originated from Western Europe (40.4%) and the United States (35.4%). The research productivity compared to population and the GNIPC was found to be higher for Canada and Oceania compared to the United States and Western Europe. The rate of increase of the total published research product (number of published articles multiplied by the impact factor) was higher in the United States and Europe. The total research contribution of Asia, Eastern Europe, Central and Latin America, and Africa regarding the number of published articles was notably very low (approximately 8%). CONCLUSIONS: The data suggest that there was a significant research activity in the field of respiratory medicine during the studied period. Although leaders of production of respiratory medicine research were from Western Europe and the United States, Canada, and Oceania had the best performance after adjustment for population and GNIPC. PMID- 16354873 TI - A 38-year-old man with HIV infection and subacute onset of cough and dyspnea. PMID- 16354872 TI - Infection of human papillomavirus type 18 and p53 codon 72 polymorphism in lung cancer patients from India. AB - STUDY OBJECTIVES: Infection with specific high-risk HPV types 16 and 18 and polymorphism of p53 codon 72 has been strongly associated with the genesis of various neoplasms in humans, but such study in lung cancer is limited and the results are controversial. In India, the role of these two factors has been strongly implicated in cervical and other cancers, but the occurrence of HPV or p53 codon 72 polymorphism has not been examined in lung cancer, which is the most common cause of cancer-related death in India. DESIGN AND PATIENTS: A total of 40 tumor biopsy specimens from advanced lung cancer patients and blood samples from 40 matching control subjects were obtained for the analysis of high-risk HPV types 16 and 18 infection and p53 codon 72 polymorphism by polymerase chain reaction. RESULTS: Only HPV type 18 was detected in 5% (2 of 40 lung cancer patients), but no other HPV could be detected. A significantly increased frequency of Arg/Arg homozygotes was observed in patients with advanced lung cancer when compared to that of control subjects (p = 0.004; odds ratio, 5.13; 95% confidence interval, 1.59 to 17.26). However, no significant correlation could be made between p53 polymorphism and different clinical stages, except for advanced stage IV patients, who showed a higher proportion of Arg/Pro heterozygous genotype. CONCLUSIONS: HPV detected in a small proportion of lung cancer patients in India demonstrated an exclusive prevalence of HPV type 18, and there was a significantly higher frequency of p53 Arg/Arg genotype when compared to that of control subjects. Observation of a shorter duration of symptoms (< or = 4 months) in as many as 78% (seven of nine stage IV patients) with Arg/Pro genotype may be an indication that lung cancer patients with the heterozygous p53 genotype are more susceptible to early progression. PMID- 16354874 TI - A 24-year-old woman with bilateral pulmonary infiltrates, pericardial effusion, and bilateral pleural effusions. PMID- 16354875 TI - Difference between dosimeter and tidal breathing methacholine challenge: contributions of dose and deep inspiration bronchoprotection. AB - BACKGROUND: Two bronchoprovocation methods are widely used. Compared to the tidal breathing method, the dosimeter method delivers approximately half the dose and involves five deep inhalations. Both the lower dose and the bronchoprotective deep inhalations contribute to the lesser airway response of the dosimeter. OBJECTIVE: To determine the relative role of dose and deep inspiration in the difference between the two methods. METHODS: Subjects with asthma (n = 24) underwent three methacholine challenges: a dosimeter challenge, a 2-min tidal breathing challenge (twice the dose), and a modified 2-min tidal breathing challenge (twice the dose plus five deep inhalations). RESULTS: The dosimeter method produced a nonsignificantly lower response than the modified tidal breathing method (p = 0.14). Both deep inhalation methods produced significantly less response than did the standard tidal breathing method (p = 0.011). In the 12 subjects with the most mild airway hyperresponsiveness (AHR), the differences between the deep inhalation method and the tidal breathing method were greater (p = 0.007). By contrast, deep inhalations produced no effect in the 12 subjects with greater AHR; the two tidal breathing methods produced identical results, while the dosimeter produced less response than either (p = 0.033). Six current asthmatics with mild airway responsiveness (tidal breathing method) had negative dosimeter methacholine challenge results. CONCLUSIONS: In subjects with moderate airway responsiveness, the difference between the methods is due to the difference in dose, whereas in subjects with mild AHR, deep inhalations had a large effect overwhelming the dose effect and producing false-negative methacholine challenge results in 25% of the subjects. PMID- 16354876 TI - Simultaneous donor marrow cell transplantation with reduced intensity conditioning prevents tracheal allograft obliteration in a bronchiolitis obliterans murine model. AB - STUDY OBJECTIVES: Prolonged survival of transplanted kidney or liver allografts has been associated with prolonged chimerism resulting from donor-origin leukocytes carried within the allograft parenchyma. The present study, performed in a murine model, examined whether simultaneous administration of donor bone marrow cells after reduced intensity conditioning allows acceptance of heterotopic tracheal allografts and prevents the formation of the airway fibroproliferative lesion, which mimics bronchiolitis obliterans (BO). METHODS: Allogeneic tracheal allografts from C57BL/6 mice were grafted subcutaneously into BALB/c mice (n = 6) [day 0]. Conditioning consisted of total lymphoid irradiation (200 cGy) at day - 1, donor marrow cells (3 x 10(7)) administered IV on day 0, intraperitoneal cyclophosphamide (200 mg/kg) on day 1 to eliminate alloreactive marrow cells, followed by a repeated dose of donor marrow cells on day 2. Control groups consisted of one group (n = 4) that underwent similar conditioning without donor marrow cells, and another group (n = 4) that received syngeneic BALB/c marrow cells. None of these groups were administered maintenance immunosuppression. Grafts were harvested and histopathology findings were evaluated semiquantitatively at day 28, day 55, and day 95. RESULTS: Tracheal allografts from donor marrow cell recipients still maintained a patent airway with intact airway epithelium at 95 days after transplant. However, grafts from control animals not receiving donor marrow cells or mice administered syngeneic marrow cells had lumen obliteration by 28 days after transplant. Chimerism in animals receiving allogeneic bone marrow was confirmed. Graft vs host disease did not develop in animals receiving allogeneic marrow cells. CONCLUSIONS: Further investigation may verify this approach to be applicable for the prevention of posttransplantation BO. PMID- 16354877 TI - T-helper type 1/T-helper type 2 balance in malignant pleural effusions compared to tuberculous pleural effusions. AB - STUDY OBJECTIVES: Malignant and tuberculous pleurisies are two major causes of lymphocyte-dominant pleurisy. Several studies have already reported that tuberculous pleurisy is a T-helper type 1(Th1)-dominant disease. In this study, we sought to examine the Th1/T-helper type 2 (Th2) balance, especially focusing on the polarizing status of T-cells to Th1/Th2 in malignant pleural effusions by measuring cytokines in pleural effusions and by evaluating the polarizing status of T-cells on the point of stimulation with interleukin (IL)-12 and/or IL-18. Furthermore, we evaluated inhibitors of interferon (IFN)-gamma production in effusions to rule out the possibility of direct inhibition of T-cell polarization. PATIENTS: Effusion samples were collected from 19 patients with malignant pleurisy caused by lung cancer and from 7 patients with tuberculous pleurisy. MEASUREMENTS: Concentrations of pleural fluid IFN-gamma, IL-12, and IL 4 were measured. IFN-gamma production of T-cells enriched from malignant pleural effusions in the presence of IL-12 and/or IL-18 was also examined. We further compared the inhibitory activity of malignant pleural effusions against IFN-gamma production and analyzed the expression of T-cell immunoglobulin mucin, mucin domain (Tim-3), a Th1-specific molecule in pleural fluid T-cells. RESULTS: Although malignant pleural effusions showed low levels of Th1 and Th2 cytokines and ratios of IFN-gamma and IL-12 to IL-4 were low, isolated T-cells produced a significant level of IFN-gamma in the presence of IL-12 and IL-18. Soluble factors were not found to inhibit IFN-gamma production in malignant pleural effusions. In tuberculous pleural effusion, ratios of IFN-gamma and IL-12 to IL-4 were significantly higher, and T-cells showed the expression of Tim-3 messenger RNA. CONCLUSIONS: We confirmed that T-cells in the malignant pleural effusions are mainly naive or not definitely polarized to Th1. Moreover, malignant tumor does not actively distort the cytokine condition through production of soluble inhibitors within effusions. The present study indicates that antitumor immunity may be enhanced by restored IFN-gamma activity through combination of IL-12 and IL-18, and that it will lead to new therapies for malignant effusion. PMID- 16354878 TI - Heat decreases formoterol delivery. AB - STUDY OBJECTIVES: Based on anecdotal reports of formoterol aggregating in mailboxes in the summer in Arizona, we examined the effect of heat on formoterol as well as on drug delivery. DESIGN: Formoterol capsules in original blister packaging were heated to 40 to 70 degrees C (104 to 158 degrees F) for 3 h and at 70 degrees C (158 degrees F) for 15 to 180 min. Capsules were removed from packaging, and a vacuum setup was used to dispense the formoterol into a filter using the device provided by the manufacturer. The weights of the capsule predispensation and postdispensation were measured to calculate drug delivery. Measurements were compared to those of capsules not exposed to heat. For comparison, tiotropium and a combination of fluticasone propionate and salmeterol (Advair; GlaxoSmithKline; Research Triangle Park, NC) were similarly tested. RESULTS: Visual inspection of the heated capsules revealed gross distortion as well as visible clumping of formoterol at the higher temperatures. The mean (+/- SEM) change in the weights of capsules that underwent heating were significantly less than those obtained from capsules that had not been heated (mean change after heating for 3 h at 70 degrees C, 2.3 +/- 0.7 vs 24.7 +/- 0.6 mg, respectively; p < 0.001), indicating decreased formoterol delivery. Heat produced a dose-responsive and time-responsive decrease in formoterol delivery. One of six capsules that were subjected to temperatures as low as 40 degrees C (104 degrees F) for 3 h had decreased delivery, and three of six capsules subjected to a temperature of 70 degrees C (158 degrees F) for times as short as 30 min decreased delivery. In contrast, neither tiotropium nor fluticasone propionate/salmeterol delivery was decreased by heating for up to 3 h at 70 degrees C (158 degrees F). Thermometers placed in mailboxes or in car windows in mid-summer in Arizona (approximate outside temperature, 110 degrees F [43 degrees C]) exceeded 70 degrees C (158 degrees F). CONCLUSIONS: These data demonstrate that the exposure of formoterol to heat decreases drug delivery and that caution should be used when mailing, transporting or storing formoterol. PMID- 16354879 TI - Influence of antiinflammatory drugs (methylprednisolone and diclofenac sodium) on experimental pleurodesis induced by silver nitrate or talc. AB - STUDY OBJECTIVE: To determine whether the administration of antiinflammatory drugs interferes with experimental pleurodesis induced by silver nitrate or talc. STUDY DESIGN: Two groups of 30 white New Zealand rabbits were scheduled to receive an intrapleural injection of 0.5% silver nitrate or 400 mg/kg of talc. Each group was further classified into three subgroups (10 animals each), which received the following: (subgroup 1) the sclerosing agent only, (subgroup 2) the sclerosing agent plus 1 mg/kg of methylprednisolone, and (subgroup 3) the sclerosing agent plus 1.1 mg/kg of diclofenac sodium. The antiinflammatory agents were administered IM 24 h before the sclerosing agent and daily during the first week, followed by once-weekly injections until death at 28 days. At this time, the pleural cavity was macroscopically evaluated, and samples of pleura and lungs were collected for further microscopic examination. MEASUREMENTS AND RESULTS: The degree of pleural adhesions was higher after silver nitrate administration (p = 0.019). No reduction in the adhesions was observed after administering antiinflammatory drugs to this group (p > 0.05). Conversely, the adhesion score was significantly reduced after administration of both prednisolone (p = 0.028) and diclofenac (p = 0.032) to the animals that received talc. Administration of the antiinflammatory agents did not influence microscopic pleural or lung changes induced by silver nitrate or talc. CONCLUSION: These results show that the sustained systemic administration of antiinflammatory agents (steroidal or nonsteroidal) reduces the degree of pleural adhesions in animals with talc induced pleurodesis but does not affect silver nitrate-induced pleurodesis. Extrapolation of these results to humans suggests that the use of antiinflammatory drugs should be avoided in patients with talc-induced pleurodesis and that appropriate clinical studies with silver nitrate should be conducted in patients chronically treated with these antiinflammatory agents. PMID- 16354880 TI - Concurrent robotic hybrid revascularization using an enhanced operative suite. AB - Hybrid myocardial revascularization combines coronary surgery with percutaneous intervention as an alternative therapy for ischemic heart disease. The order and sequence of the hybrid approach is not yet clearly defined. We report on the benefits of an enhanced surgical suite equipped with a carbon fiber operating table and digital C-arm for robotic-assisted hybrid revascularization in a single operative sequence. To our knowledge, this is the first reported case of concurrent robotic-assisted hybrid revascularization utilizing an enhanced operative suite. PMID- 16354881 TI - Detection of hypoventilation. PMID- 16354882 TI - Aging and induced-sputum cells. PMID- 16354883 TI - A modified percutaneous tracheostomy technique without bronchoscopic guidance: a note of concern. PMID- 16354884 TI - Ribavarin should be tested in clinical trials in combination with other antiviral agents for severe acute respiratory syndrome. PMID- 16354885 TI - "Frequent fliers" do not receive a free trip in the emergency department. PMID- 16354886 TI - Obesity and migraine: a population study. AB - OBJECTIVE: To assess the influence of body mass index (BMI) on the prevalence, attack frequency, and clinical features of migraine. METHODS: In a population based telephone interview study, the authors gathered information on headache, height, and weight. The 30,215 participants were divided into five categories, based on BMI: 1, underweight (< 18.5), normal weight (18.5 to 24.9), overweight (25 to 29.9), obese (30 to 24.9), and morbidly obese (> or = 35). Migraine prevalence and modeled headache features were assessed as a function of BMI, adjusting by covariates (age, sex, marital status, income, medical treatment, depression). RESULTS: Subjects were predominantly female (65% female) and in middle life (mean age 38.4). BMI group was not associated with the prevalence of migraine, but was associated with the frequency of headache attacks. In the normal weight group, 4.4% had 10 to 15 headache days per month, increasing to 5.8% of the overweight (odds ratio [OR] = 1.3), 13.6% of the obese (OR = 2.9), and 20.7% of the morbidly obese (OR = 5.7). The proportion of subjects with severe headache pain increased with BMI, doubling in the morbidly obese relative to the normally weighted (OR = 1.9). Similar significant associations were demonstrated with BMI category for disability, photophobia, and phonophobia. CONCLUSION: Though migraine prevalence is not associated with body mass index, attack frequency, severity, and clinical features of migraine increase with body mass index group. PMID- 16354887 TI - Images in clinical medicine. Urinary calcium oxalate crystals in ethylene glycol intoxication. PMID- 16354888 TI - Glimpses of Guantanamo--medical ethics and the war on terror. PMID- 16354889 TI - Influenza--interpandemic as well as pandemic disease. PMID- 16354890 TI - A trial of contraceptive methods in women with systemic lupus erythematosus. AB - BACKGROUND: The effects of estrogen-containing contraceptives on disease activity in women with systemic lupus erythematosus have not been determined. METHODS: We conducted a single-blind clinical trial involving 162 women with systemic lupus erythematosus who were randomly assigned to combined oral contraceptives, a progestin-only pill, or a copper intrauterine device (IUD). Disease activity was assessed at 0, 1, 2, 3, 6, 9, and 12 months according to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). The primary outcome was global disease activity, which we estimated by measuring the area under the SLEDAI curve. Secondary outcomes included the maximum SLEDAI score, change in SLEDAI score, incidence of lupus flares, median time to first flare, systemic lupus erythematosus treatment, and adverse events. The results were analyzed by the intention-to-treat method. RESULTS: At baseline, all demographic features and disease characteristics were similar in the three groups. The mean (+/-SD) SLEDAI score was 6.1+/-5.6 in the group assigned to combined oral contraceptives, 6.4+/ 4.6 in the group assigned to the progestin-only pill, and 5.0+/-5.3 in the group assigned to the IUD (54 patients in each group) (P=0.36). Disease activity remained mild and stable in all groups throughout the trial. There were no significant differences among the groups during the trial in global or maximum disease activity, incidence or probability of flares, or medication use. The median time to the first flare was three months in all groups. Thromboses occurred in four patients (two in each of the two groups receiving hormones), and severe infections were more frequent in the IUD group. One patient receiving combined oral contraceptives died from amoxicillin-related severe neutropenia. CONCLUSIONS: Global disease activity, maximum SLEDAI score, incidence of flares, time to first flare, and incidence of adverse events were similar among women with systemic lupus erythematosus, irrespective of the type of contraceptive they were using. PMID- 16354891 TI - Combined oral contraceptives in women with systemic lupus erythematosus. AB - BACKGROUND: Oral contraceptives are rarely prescribed for women with systemic lupus erythematosus, because of concern about potential negative side effects. In this double-blind, randomized, noninferiority trial, we prospectively evaluated the effect of oral contraceptives on lupus activity in premenopausal women with systemic lupus erythematosus. METHODS: A total of 183 women with inactive (76 percent) or stable active (24 percent) systemic lupus erythematosus at 15 U.S. sites were randomly assigned to receive either oral contraceptives (triphasic ethinyl estradiol at a dose of 35 microg plus norethindrone at a dose of 0.5 to 1 mg for 12 cycles of 28 days each; 91 women) or placebo (92 women) and were evaluated at months 1, 2, 3, 6, 9, and 12. Subjects were excluded if they had moderate or high levels of anticardiolipin antibodies, lupus anticoagulant, or a history of thrombosis. RESULTS: The primary end point, a severe lupus flare, occurred in 7 of 91 subjects receiving oral contraceptives (7.7 percent) as compared with 7 of 92 subjects receiving placebo (7.6 percent). The 12-month rates of severe flare were similar: 0.084 for the group receiving oral contraceptives and 0.087 for the placebo group (P=0.95; upper limit of the one sided 95 percent confidence interval for this difference, 0.069, which is within the prespecified 9 percent margin for noninferiority). Rates of mild or moderate flares were 1.40 flares per person-year for subjects receiving oral contraceptives and 1.44 flares per person-year for subjects receiving placebo (relative risk, 0.98; P=0.86). In the group that was randomized to receive oral contraceptives, there was one deep venous thrombosis and one clotted graft; in the placebo group, there was one deep venous thrombosis, one ocular thrombosis, one superficial thrombophlebitis, and one death (after cessation of the trial). CONCLUSIONS: Our study indicates that oral contraceptives do not increase the risk of flare among women with systemic lupus erythematosus whose disease is stable. PMID- 16354892 TI - Influenza-associated deaths among children in the United States, 2003-2004. AB - BACKGROUND: Although influenza is common among children, pediatric mortality related to laboratory-confirmed influenza has not been assessed nationally. METHODS: During the 2003-2004 influenza season, we requested that state health departments report any death associated with laboratory-confirmed influenza in a U.S. resident younger than 18 years of age. Case reports, medical records, and autopsy reports were reviewed, and available influenza-virus isolates were analyzed at the Centers for Disease Control and Prevention. RESULTS: One hundred fifty-three influenza-associated deaths among children were reported by 40 state health departments. The median age of the children was three years, and 96 of them (63 percent) were younger than five years old. Forty-seven of the children (31 percent) died outside a hospital setting, and 45 (29 percent) died within three days after the onset of illness. Bacterial coinfections were identified in 24 of the 102 children tested (24 percent). Thirty-three percent of the children had an underlying condition recognized to increase the risk of influenza-related complications, and 20 percent had other chronic conditions; 47 percent had previously been healthy. Chronic neurologic or neuromuscular conditions were present in one third. The mortality rate was highest among children younger than six months of age (0.88 per 100,000 children; 95 percent confidence interval, 0.52 to 1.39 per 100,000). CONCLUSIONS: A substantial number of influenza associated deaths occurred among U.S. children during the 2003-2004 influenza season. High priority should be given to improvements in influenza-vaccine coverage and improvements in the diagnosis and treatment of influenza to reduce childhood mortality from influenza. PMID- 16354893 TI - Atrial overdrive pacing for the obstructive sleep apnea-hypopnea syndrome. AB - BACKGROUND: The role of atrial overdrive pacing (AOP) in sleep apnea remains uncertain. We prospectively evaluated the effect of AOP after 24 hours and after one month in patients with the obstructive sleep apnea-hypopnea syndrome and compared it with the use of nasal continuous positive airway pressure (n-CPAP). METHODS: We studied 16 patients with a moderate or severe case of the obstructive sleep apnea-hypopnea syndrome (baseline mean apnea-hypopnea index, 49) and normal left ventricular systolic function in whom a dual-chamber pacemaker had been implanted. After 48 hours, the patients were randomly assigned to AOP (pacing at 15 bpm above the spontaneous mean nocturnal heart rate) or backup atrial pacing (pacing at a heart rate below 40 bpm); the latter group began n-CPAP therapy one day later. After one month, the two groups switched therapies and were followed for an additional month. Polysomnographic studies were performed at baseline, on the first night after randomization, at crossover, and at the end of the study. RESULTS: During AOP, no significant changes were observed in any of the respiratory variables measured. The change in the apnea-hypopnea index at one month with AOP was +0.2 (95 percent confidence interval, -2.7 to +2.3; P=0.87). In contrast, all variables improved significantly after one month of n-CPAP (change in the apnea-hypopnea index, -46.3; 95 percent confidence interval, -56.2 to -36.5; P<0.001). CONCLUSIONS: Nasal continuous positive airway pressure therapy is highly effective for the treatment of the obstructive sleep apnea hypopnea syndrome, whereas AOP has no significant effect. PMID- 16354894 TI - Clinical practice. Hirsutism. PMID- 16354895 TI - Images in clinical medicine. Pemphigus foliaceus. PMID- 16354896 TI - Case records of the Massachusetts General Hospital. Case 38-2005. A 29-year-old pregnant woman with the nephrotic syndrome and hypertension. PMID- 16354897 TI - Oral contraceptives in systemic lupus erythematosus--a tough pill to swallow? PMID- 16354898 TI - Can sleep apnea be treated without modifying anatomy? PMID- 16354899 TI - Appropriate use of artificial nutrition and hydration--fundamental principles and recommendations. PMID- 16354900 TI - Circulating endothelial progenitor cells. PMID- 16354901 TI - Paget's disease and bisphosphonates. PMID- 16354902 TI - Alendronate and parathyroid hormone. PMID- 16354903 TI - A sad day for science at the FDA. PMID- 16354904 TI - Sudden reversible osmotic lens damage ("sugar cracks") after initiation of metformin. PMID- 16354905 TI - Oxidants off the hook? PMID- 16354906 TI - When T cells get old. AB - Why is vaccination against infectious diseases less effective in older patients than in younger ones? Why do the elderly suffer from more frequent and severe infectious episodes than the young? The answer to both these questions is immunosenescence--the poorly defined changes that occur in the immune system as a result of the disrupted performance of multiple components of immunity. Presentations from a recent workshop, which are summarized here, examined these questions and provided some insights from the perspective of improving vaccination strategies among the elderly. PMID- 16354907 TI - IgG-assisted age-dependent clearance of Alzheimer's amyloid beta peptide by the blood-brain barrier neonatal Fc receptor. AB - The role of blood-brain barrier (BBB) transport in clearance of amyloid beta peptide (Abeta) by Abeta immunotherapy is not fully understood. To address this issue, we studied the effects of peripherally and centrally administered Abeta specific IgG on BBB influx of circulating Abeta and efflux of brain-derived Abeta in APPsw(+/-) mice, a model that develops Alzheimer's disease-like amyloid pathology, and wild-type mice. Our data show that anti-Abeta IgG blocks the BBB influx of circulating Abeta in APPsw(+/-) mice and penetrates into the brain to sequester brain Abeta. In young mice, Abeta-anti-Abeta complexes were cleared from brain to blood by transcytosis across the BBB via the neonatal Fc receptor (FcRn) and the low-density lipoprotein receptor-related protein (LRP), whereas in older mice, there was an age-dependent increase in FcRn-mediated IgG-assisted Abeta BBB efflux and a decrease in LRP-mediated clearance of Abeta-anti-Abeta complexes. Inhibition of the FcRn pathway in older APPsw(+/-) mice blocked clearance of endogenous Abeta40/42 by centrally administered Abeta immunotherapy. Moreover, deletion of the FcRn gene in wild-type mice inhibited clearance of endogenous mouse Abeta40/42 by systemically administered anti-Abeta. Our data suggest that the FcRn pathway at the BBB plays a crucial role in IgG-assisted Abeta removal from the aging brain. PMID- 16354908 TI - Short-term regulation of information processing at the corticoaccumbens synapse. AB - In relation to expectation and delivery of reward, pyramidal neurons of the prefrontal cortex either switch from a single spiking mode to transient phasic bursting, or gradually increase their sustained tonic activity. Here, we examined how switching between firing modes affects information processing at the corticoaccumbens synapse. We report that increasing presynaptic firing frequency in a tonic manner either depresses or facilitates synaptic transmission, depending on initial probability of release. In contrast, repeated bursts of stimulation of cortical afferents trigger a new form of short-term potentiation of synaptic transmission (RB-STP) in the nucleus accumbens (NAc). RB-STP involves the regulation of axonal excitability mediated by 4-AP-sensitive potassium channels in afferent cortical neurons. Thus, in a tonic mode, information flow is tightly controlled by regulatory mechanisms at the level of presynaptic terminals, whereas switching to a bursting mode reliably enhances efficacy of information processing for all cortical afferents to NAc neurons. PMID- 16354909 TI - Extrasynaptic GABAA receptors of thalamocortical neurons: a molecular target for hypnotics. AB - Among hypnotic agents that enhance GABAA receptor function, etomidate is unusual because it is selective for beta2/beta3 compared with beta1 subunit-containing GABAA receptors. Mice incorporating an etomidate-insensitive beta2 subunit (beta(2N265S)) revealed that beta2 subunit-containing receptors mediate the enhancement of slow-wave activity (SWA) by etomidate, are required for the sedative, and contribute to the hypnotic actions of this anesthetic. Although the anatomical location of the beta2-containing receptors that mediate these actions is unknown, the thalamus is implicated. We have characterized GABAA receptor mediated neurotransmission in thalamic nucleus reticularis (nRT) and ventrobasalis complex (VB) neurons of wild-type, beta2(-/-), and beta(2N265S) mice. VB but not nRT neurons exhibit a large GABA-mediated tonic conductance that contributes approximately 80% of the total GABAA receptor-mediated transmission. Consequently, although etomidate enhances inhibition in both neuronal types, the effect of this anesthetic on the tonic conductance of VB neurons is dominant. The GABA-enhancing actions of etomidate in VB but not nRT neurons are greatly suppressed by the beta(2N265S) mutation. The hypnotic THIP (Gaboxadol) induces SWA and at low, clinically relevant concentrations (30 nM to 3 microM) increases the tonic conductance of VB neurons, with no effect on VB or nRT miniature IPSCs (mIPSCs) or on the holding current of nRT neurons. Zolpidem, which has no effect on SWA, prolongs VB mIPSCs but is ineffective on the phasic and tonic conductance of nRT and VB neurons, respectively. Collectively, these findings suggest that enhancement of extrasynaptic inhibition in the thalamus may contribute to the distinct sleep EEG profiles of etomidate and THIP compared with zolpidem. PMID- 16354910 TI - Mecp2 deficiency disrupts norepinephrine and respiratory systems in mice. AB - Rett syndrome is a severe X-linked neurological disorder in which most patients have mutations in the methyl-CpG binding protein 2 (MECP2) gene and suffer from bioaminergic deficiencies and life-threatening breathing disturbances. We used in vivo plethysmography, in vitro electrophysiology, neuropharmacology, immunohistochemistry, and biochemistry to characterize the consequences of the MECP2 mutation on breathing in wild-type (wt) and Mecp2-deficient (Mecp2-/y) mice. At birth, Mecp2-/y mice showed normal breathing and a normal number of medullary neurons that express tyrosine hydroxylase (TH neurons). At approximately 1 month of age, most Mecp2-/y mice showed respiratory cycles of variable duration; meanwhile, their medulla contained a significantly reduced number of TH neurons and norepinephrine (NE) content, even in Mecp2-/y mice that showed a normal breathing pattern. Between 1 and 2 months of age, all unanesthetized Mecp2-/y mice showed breathing disturbances that worsened until fatal respiratory arrest at approximately 2 months of age. During their last week of life, Mecp2-/y mice had a slow and erratic breathing pattern with a highly variable cycle period and frequent apneas. In addition, their medulla had a drastically reduced number of TH neurons, NE content, and serotonin (5-HT) content. In vitro experiments using transverse brainstem slices of mice between 2 and 3 weeks of age revealed that the rhythm produced by the isolated respiratory network was irregular in Mecp2-/y mice but could be stabilized with exogenous NE. We hypothesize that breathing disturbances in Mecp2-/y mice, and probably Rett patients, originate in part from a deficiency in noradrenergic and serotonergic modulation of the medullary respiratory network. PMID- 16354911 TI - A C-terminal domain directs Kv3.3 channels to dendrites. AB - Pyramidal neurons of the electrosensory lateral line lobe (ELL) of Apteronotus leptorhynchus express Kv3-type voltage-gated potassium channels that give rise to high-threshold currents at the somatic and dendritic levels. Two members of the Kv3 channel family, AptKv3.1 and AptKv3.3, are coexpressed in these neurons. AptKv3.3 channels are expressed at uniformly high levels in each of four ELL segments, whereas AptKv3.1 channels appear to be expressed in a graded manner with higher levels of expression in segments that process high-frequency electrosensory signals. Immunohistochemical and recombinant channel expression studies show a differential distribution of these two channels in the dendrites of ELL pyramidal neurons. AptKv3.1 is concentrated in somas and proximal dendrites, whereas AptKv3.3 is distributed throughout the full extent of the large dendritic tree. Recombinant channel expression of AptKv3 channels through in vivo viral injections allowed directed retargeting of AptKv3 subtypes over the somadendritic axis, revealing that the sequence responsible for targeting channels to distal dendrites lies within the C-terminal domain of the AptKv3.3 protein. The targeting domain includes a consensus sequence predicted to bind to a PDZ (postsynaptic density-95/Discs large/zona occludens-1)-type protein-protein interaction motif. These findings reveal that different functional roles for Kv3 potassium channels at the somatic and dendritic level of a sensory neuron are attained through specific targeting that selectively distributes Kv3.3 channels to the dendritic compartment. PMID- 16354912 TI - Suppression of cyclin-dependent kinase 5 activation by amyloid precursor protein: a novel excitoprotective mechanism involving modulation of tau phosphorylation. AB - Alzheimer's disease is cytopathologically characterized by loss of synapses and neurons, neuritic amyloid plaques consisting of beta-amyloid (Abeta) peptides, and neurofibrillary tangles consisting of hyperphosphorylated tau protein in susceptible brain regions. Abeta, which triggers a cascade of pathogenic events including tau phosphorylation and neuronal excitotoxicity, is proteolytically derived from beta-amyloid precursor protein (APP); the pathological and physiological functions of APP, however, remain undefined. Here we demonstrate that the level of tau phosphorylation in cells and brains deficient in APP is significantly higher than that in wild-type controls, resulting from activation of cyclin-dependent kinase 5 (CDK5) but not glycogen synthase kinase 3, the two major tau kinases. In addition, we show that overexpression of APP or its non amyloidogenic homolog amyloid precursor-like protein 1 suppresses both basal and stress-induced CDK5 activation. The ectodomain of APP, sAPPalpha, is responsible for inhibiting CDK5 activation. Furthermore, neurons derived from APP-deficient mice exhibit reduced metabolism and survival rates and are more susceptible to excitotoxic glutamate-induced apoptosis. These neurons also manifest significant defects in neurite outgrowth compared with neurons from the wild-type littermates. The observed neuronal excitotoxicity/apoptosis is mediated through a mechanism involving CDK5 activation. Our study defines a novel neuroprotective function for APP in preventing tau hyperphosphorylation via suppressing overactivation of CDK5. We suggest that CDK5 activation, through a calcium/calpain/p25 pathway, plays a key role in neuronal excitotoxicity and represents an underlying mechanism for the physiological functions of APP. PMID- 16354913 TI - GABAA receptor-mediated tonic inhibition in thalamic neurons. AB - Tonic GABAA receptor-mediated inhibition is typically generated by delta subunit containing extrasynaptic receptors. Because the delta subunit is highly expressed in the thalamus, we tested whether thalamocortical (TC) neurons of the dorsal lateral geniculate nucleus (dLGN) and ventrobasal complex exhibit tonic inhibition. Focal application of gabazine (GBZ) (50 microM) revealed the presence of a 20 pA tonic current in 75 and 63% of TC neurons from both nuclei, respectively. No tonic current was observed in GABAergic neurons of the nucleus reticularis thalami (NRT). Bath application of 1 microM GABA increased tonic current amplitude to approximately 70 pA in 100% of TC neurons, but it was still not observed in NRT neurons. In dLGN TC neurons, the tonic current was sensitive to low concentrations of the delta subunit-specific receptor agonists allotetrahydrodeoxycorticosterone (100 nM) and 4,5,6,7-tetrahydroisoxazolo[5,4-c] pyridin-3-ol (THIP) (100 nM) but insensitive to the benzodiazepine flurazepam (5 microM). Bath application of low concentrations of GBZ (25-200 nM) preferentially blocked the tonic current, whereas phasic synaptic inhibition was primarily maintained. Under intracellular current-clamp conditions, the preferential block of the tonic current with GBZ led to a small depolarization and increase in input resistance. Using extracellular single-unit recordings, block of the tonic current caused the cessation of low-threshold burst firing and promoted tonic firing. Enhancement of the tonic current by THIP hyperpolarized TC neurons and promoted burst firing. Thus, tonic current in TC neurons generates an inhibitory tone. Its modulation contributes to the shift between different firing modes, promotes the transition between different behavioral states, and predisposes to absence seizures. PMID- 16354914 TI - Abnormalities of neurogenesis in the R6/2 mouse model of Huntington's disease are attributable to the in vivo microenvironment. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative condition characterized by movement disorders, psychiatric disturbance, and cognitive decline. There are no treatments to halt or reverse the disease. Mammalian neurogenesis persists into adulthood in the subventricular zone (SVZ) and dentate gyrus (DG) of the hippocampus. In 2001, our laboratory published the hypothesis that neurogenesis is impaired in neurodegenerative diseases and that this may contribute to disease progression. Since then, it has been shown that neurogenesis is reduced in the DG of transgenic HD mice but increased in the SVZ of HD patients. We sought to characterize neurogenesis further. We found that, in the DG of the transgenic R6/2 mouse model of HD, newborn cell proliferation and morphology, but not differentiation or survival, was compromised. In R6/2 mice, neurogenesis failed to upregulate in the DG in response to seizures. Basal SVZ neurogenesis was similar between R6/2 mice and their wild-type littermates. There was no difference in the in vitro growth of adult neural precursor cells (NPCs) between genotypes. These results suggest that abnormal neurogenesis in the R6/2 mouse is not attributable to an intrinsic impairment of the NPC itself but is attributable to the environment in which the cell is located. PMID- 16354915 TI - Few CaV1.3 channels regulate the exocytosis of a synaptic vesicle at the hair cell ribbon synapse. AB - Hearing relies on faithful sound coding at hair cell ribbon synapses, which use Ca2+-triggered glutamate release to signal with submillisecond precision. Here, we investigated stimulus-secretion coupling at mammalian inner hair cell (IHC) synapses to explore the mechanisms underlying this high temporal fidelity. Using nonstationary fluctuation analysis on Ca2+ tail currents, we estimate that IHCs contain approximately 1700 Ca2+ channels, mainly of CaV1.3 type. We show by immunohistochemistry that the CaV1.3 Ca2+ channels are localized preferentially at the ribbon-type active zones of IHCs. We argue that each active zone holds approximately 80 Ca2+ channels, of which probably <10 open simultaneously during physiological stimulation. We then manipulated the Ca2+ current by primarily changing single-channel current or open-channel number. Effects on exocytosis of the readily releasable vesicle pool (RRP) were monitored by membrane capacitance recordings. Consistent with the high intrinsic Ca2+ cooperativity of exocytosis, RRP exocytosis changed nonlinearly with the Ca2+ current when varying the single channel current. In contrast, the apparent Ca2+ cooperativity of RRP exocytosis was close to unity when primarily manipulating the number of open channels. Our findings suggest a Ca2+ channel-release site coupling in which few nearby CaV1.3 channels impose high nanodomain [Ca2+] on release sites in IHCs during physiological stimulation. We postulate that the IHC ribbon synapse uses this Ca2+ nanodomain control of exocytosis to signal with high temporal precision already at low sound intensities. PMID- 16354916 TI - Distinct roles for different Homer1 isoforms in behaviors and associated prefrontal cortex function. AB - Homer1 mutant mice exhibit behavioral and neurochemical abnormalities that are consistent with an animal model of schizophrenia. Because the Homer1 gene encodes both immediate early gene (IEG) and constitutively expressed (CC) gene products, we used the local infusion of adeno-associated viral vectors carrying different Homer1 transcriptional variants into the prefrontal cortex (PFC) to distinguish between the roles for IEG and CC Homer1 isoforms in the "schizophrenia-like" phenotype of Homer1 mutant mice. PFC overexpression of the IEG Homer1 isoform Homer1a reversed the genotypic differences in behavioral adaptation to repeated stress, whereas overexpression of the constitutively expressed Homer1 isoform Homer1c reversed the genotypic differences in sensorimotor and cognitive processing, as well as cocaine behavioral sensitivity. Homer1a overexpression did not influence PFC basal glutamate content but blunted the glutamate response to cocaine in wild-type mice. In contrast, Homer1c overexpression reversed the genotypic difference in PFC basal glutamate content and enhanced cocaine-induced elevations in glutamate. These data demonstrate active and distinct roles for Homer1a and Homer1c isoforms in the PFC in the mediation of behavior, in the maintenance of basal extracellular glutamate, and in the regulation of PFC glutamate release relevant to schizophrenia and stimulant abuse comorbidity. PMID- 16354917 TI - Brn3a-expressing retinal ganglion cells project specifically to thalamocortical and collicular visual pathways. AB - Retinal ganglion cells (RGCs) innervate several specific CNS targets serving cortical and subcortical visual pathways and the entrainment of circadian rhythms. Recent studies have shown that retinal ganglion cells express specific combinations of POU- and LIM-domain transcription factors, but how these factors relate to the subsequent development of the retinofugal pathways and the functional identity of RGCs is mostly unknown. Here, we use targeted expression of an genetic axonal tracer, tau/beta-galactosidase, to examine target innervation by retinal ganglion cells expressing the POU-domain factor Brn3a. Brn3a is expressed in RGCs innervating the principal retinothalamic/retinocollicular pathway mediating cortical vision but is not expressed in RGCs of the accessory optic, pretectal, and hypothalamic pathways serving subcortical visuomotor and circadian functions. In the thalamus, Brn3a ganglion cell fibers are primarily restricted to the outer shell of the dorsal lateral geniculate, providing new evidence for the regionalization of this nucleus in rodents. Brn3a RGC axons have a relative preference for the contralateral hemisphere, but known mediators of the laterality of RGC axons are not repatterned in the absence of Brn3a. Brn3a is coexpressed extensively with the closely related factor Brn3b in the embryonic retina, and the effects of the loss of Brn3a in retinal development are not severe, suggesting partial redundancy of function in this gene class. PMID- 16354918 TI - Neurosteroid access to the GABAA receptor. AB - GABAA receptors are a pivotal inhibitory influence in the nervous system, and modulators of the GABAA receptor are important anesthetics, sedatives, anticonvulsants, and anxiolytics. Current views of receptor modulation suggest that many exogenous drugs access and bind to an extracellular receptor domain. Using novel synthetic steroid analogs, we examined the access route for neuroactive steroids, potent GABAA receptor modulators also produced endogenously. Tight-seal recordings, in which direct aqueous drug access to receptor was prevented, demonstrated that steroids can reach the receptor either through plasma membrane lateral diffusion or through intracellular routes. A fluorescent neuroactive steroid accumulated intracellularly, but recordings from excised patches indicated that the intracellular reservoir is not necessary for receptor modulation, although it can apparently equilibrate with the plasma membrane within seconds. A membrane impermeant neuroactive steroid modulated receptor activity only when applied to the inner membrane leaflet, demonstrating that the steroid does not access an extracellular modulatory site. Thus, neuroactive steroids do not require direct aqueous access to the receptor, and membrane accumulation is required for receptor modulation. PMID- 16354919 TI - Focal and temporal release of glutamate in the mushroom bodies improves olfactory memory in Apis mellifera. AB - In contrast to vertebrates, the role of the neurotransmitter glutamate in learning and memory in insects has hardly been investigated. The reason is that a pharmacological characterization of insect glutamate receptors is still missing; furthermore, it is difficult to locally restrict pharmacological interventions. In this study, we overcome these problems by using locally and temporally defined photo-uncaging of glutamate to study its role in olfactory learning and memory formation in the honeybee, Apis mellifera. Uncaging glutamate in the mushroom bodies immediately after a weak training protocol induced a higher memory rate 2 d after training, mimicking the effect of a strong training protocol. Glutamate release before training does not facilitate memory formation, suggesting that glutamate mediates processes triggered by training and required for memory formation. Uncaging glutamate in the antennal lobes shows no effect on memory formation. These results provide the first direct evidence for a temporally and locally restricted function of glutamate in memory formation in honeybees and insects. PMID- 16354920 TI - Presynaptic homeostatic plasticity rescues long-term depression after chronic Delta 9-tetrahydrocannabinol exposure. AB - Alterations of long-term synaptic plasticity have been proposed to participate in the development of addiction. To preserve synaptic functions, homeostatic processes must be engaged after exposure to abused drugs. At the mouse cortico accumbens synapses, a single in vivo injection of Delta9-tetrahydrocannabinol (THC) suppresses endocannabinoid-mediated long-term depression. Using biochemical and electrophysiological approaches, we now report that 1 week of repeated in vivo THC treatment reduces the coupling efficiency of cannabinoid CB1 receptors (CB1Rs) to G(i/o) transduction proteins, as well as CB1R-mediated inhibition of excitatory synaptic transmission at the excitatory synapses between the prefrontal cortex and the nucleus accumbens (NAc). Nonetheless, we found that cortico-accumbens synapses unexpectedly express normal long-term depression because of a reversible switch in its underlying mechanisms. The present data show that, in THC-treated mice, long-term depression is expressed because a presynaptic mGluR2/3 (metabotropic glutamate receptor 2/3)-dependent mechanism replaces the impaired endocannabinoid system. Thus, in the NAc, a novel form of presynaptic homeostasis rescues synaptic plasticity from THC-induced deficits. PMID- 16354921 TI - Attentional selection and action selection in the ventral and orbital prefrontal cortex. AB - Different accounts of the ventral and orbital prefrontal cortex (PFv+o) have emphasized either its role in learning conditional rules for action selection or the attentional selection of behaviorally relevant stimuli. Although the accounts are not mutually exclusive, it is possible that the involvement of PFv+o in conditional action selection is a consequence of its role in selecting relevant stimuli or that its involvement in attentional selection is a consequence of the conditional rules present in many attentional paradigms. Five macaques learned a conditional action-selection task in which the difficulty of identifying the stimulus relevant for guiding action selection was varied in a simple manner by either altering its distance from the action or presenting additional distracting stimuli. Simply increasing the spatial separation between the instructing stimulus led to slower responses. Experiment 1 showed that bilateral PFv+o lesions impaired conditional action selection even when attentional demands were kept to a minimum, but there was evidence that the impairment was exacerbated by manipulating stimulus selection difficulty. Experiment 2 confirmed the importance of PFv+o for conditional action selection even when stimulus selection difficulty was minimal. Experiments 3 and 4 demonstrated that the action-selection impairment was significantly increased by making identification of the behaviorally relevant stimulus difficult. PFv+o is central to the use of conditional rules when selecting courses of action, but conditional rules are also represented in premotor and striatal regions. A special contribution of PFv+o may be initial selection of behaviorally relevant stimuli. PMID- 16354922 TI - Functional magnetic resonance imaging shows oxytocin activates brain regions associated with mother-pup bonding during suckling. AB - Oxytocin is released in the maternal brain during breastfeeding and may help strengthen the mother-infant relationship. Here, we used functional magnetic resonance imaging to determine whether oxytocin modulates brain activity in postpartum day 4-8 dams receiving suckling stimulation. During imaging sessions, dams were exposed to pup suckling before and after administration of an oxytocin receptor antagonist. Another group of dams received oxytocin alone. Changes in brain activation in response to suckling closely matched that elicited by oxytocin administration. The overlapping brain areas included the olfactory system, nucleus accumbens, insular cortex, prefrontal cortex, ventral tegmental area, cortical amygdala, and several cortical and hypothalamic nuclei. Blockade of oxytocin receptors largely attenuated activation in these regions. The data suggest that oxytocin may strengthen mother-infant bond formation partly by acting through brain areas involved in regulating olfactory discrimination, emotions, and reward. PMID- 16354923 TI - Dysregulation of receptor interacting protein-2 and caspase recruitment domain only protein mediates aberrant caspase-1 activation in Huntington's disease. AB - Caspase-1 plays a role in the pathogenesis of a variety of neurological diseases. Caspase-1 activation is an early event in models of Huntington's disease (HD). However, mechanisms regulating the activation of this apical caspase in cell death are not known. Receptor interacting protein-2 (Rip2) and caspase recruitment domain (CARD) only protein (Cop) are two CARD proteins with significant homology to the caspase-1 CARD and modulate caspase-1 activation in inflammation. Rip2 is a caspase-1 activator, and Cop is a caspase-1 inhibitor. We demonstrate in models of HD that caspase-1 activation results from dysregulation of caspase-1 activation pathways. Associated with disease progression, we detect elevation of the caspase-1 activator Rip2 and reduction of the caspase-1 inhibitor Cop. Knocking down endogenous Rip2/Cop respectively results in reduced/increased sensitivity to neurotoxic stimuli. Our data provide evidence that caspase-1-mediated cell death is regulated, at least in part, by the balance of Rip2 and Cop, and alterations of this balance may contribute to aberrant caspase-1-mediated pathogenesis in Huntington's disease. PMID- 16354924 TI - The influence of multivesicular release and postsynaptic receptor saturation on transmission at granule cell to Purkinje cell synapses. AB - The properties of a synapse are crucially dependent on whether an action potential can trigger the release of multiple vesicles at an individual release site [multivesicular release (MVR)] and whether fusion of a single vesicle leads to receptor saturation. MVR and receptor saturation both occur at some high p synapses, but it is not known whether they also occur at low p synapses. Here we examine this issue at the low p synapse between parallel fibers and Purkinje cells using the low-affinity antagonist DGG (gamma-D-glutamylglycine) to relieve AMPA receptor saturation. We find that the presence of MVR and receptor saturation at this synapse alters the calcium dependence of synaptic transmission and reduces the extent of facilitation. These findings establish that MVR and postsynaptic receptor saturation can influence transmission even at synapses with a low initial probability of release and suggest that these properties may be common at synapses in the mammalian brain. PMID- 16354925 TI - Early and late mechanisms of surround suppression in striate cortex of macaque. AB - The response of a neuron in striate cortex to an optimally configured visual stimulus is generally reduced when the stimulus is enlarged to encroach on a suppressive region that surrounds its classical receptive field (CRF). To characterize the mechanism that gives rise to this suppression, we measured its spatiotemporal tuning, its susceptibility to contrast adaptation, and its capacity for interocular transfer. Responses to an optimally configured grating confined to the CRF were strongly suppressed by annular surrounding gratings drifting at a wide range of temporal and spatial frequencies (including spatially uniform fields) that extended from well below to well above the range that drives most cortical neurons. Suppression from gratings capable of driving cortical CRFs was profoundly reduced by contrast adaptation and showed substantial interocular transfer. Suppression from stimuli that lay outside the spatiotemporal passband of most cortical CRFs was relatively stronger when the stimulus on the CRF was of low contrast, was generally insusceptible to contrast adaptation, and showed little interocular transfer. Our findings point to the existence of two mechanisms of surround suppression: one that is prominent when high-contrast stimuli drive the CRF, is orientation selective, has relatively sharp spatiotemporal tuning, is binocularly driven, and can be substantially desensitized by adaptation; the other is relatively more prominent when low contrast stimuli drive the CRF, has very broad spatiotemporal tuning, is monocularly driven, and is insusceptible to adaptation. Its character suggests an origin in the input layers of primary visual cortex, or earlier. PMID- 16354926 TI - Activity-dependent acceleration of endocytosis at a central synapse. AB - Accumulated evidence indicates the existence of rapid and slow endocytosis at many synapses. It has been proposed that rapid endocytosis is activated by intense stimulation when vesicle recycling needs to be speeded up to supply vesicles at hippocampal synapses. However, the evidence, as obtained with imaging techniques, which are somewhat indirect in indicating rapid endocytosis, is controversial. Furthermore, a slower time course of endocytosis is often found after more intense nerve activity, casting doubt on the role of rapid endocytosis at synapses. Here, we addressed this issue at a mammalian central synapse, the calyx of Held, using a capacitance measurement technique that provides a higher time resolution than imaging techniques. We found that rapid endocytosis with a time constant of approximately 1-2 s was activated during intense nerve activity. Reducing the presynaptic calcium current or buffering the intracellular calcium with EGTA significantly inhibited rapid endocytosis, suggesting that calcium triggers rapid endocytosis. During intense stimulation, rapid endocytosis retrieved up to approximately eight vesicles per second per active zone, approximately eightfold larger than reported in the hippocampus, and thus played a dominant role during and within 3 s after intense stimulation. Slow endocytosis became dominant 3 s after intense stimulation likely because of the fall of the intracellular calcium level that deactivated rapid endocytosis. These results underscore the importance of calcium-triggered rapid endocytosis, which offers the nerve terminal the plasticity to speed up vesicle cycling during intense nerve activity. PMID- 16354927 TI - Visual deprivation modifies both presynaptic glutamate release and the composition of perisynaptic/extrasynaptic NMDA receptors in adult visual cortex. AB - Use-dependent modifications of synapses have been well described in the developing visual cortex, but the ability for experience to modify synapses in the adult visual cortex is poorly understood. We found that 10 d of late-onset visual deprivation modifies both presynaptic and postsynaptic elements at the layer 4-->2/3 connection in the visual cortex of adult mice, and these changes differ from those observed in juveniles. Although visual deprivation in juvenile mice modifies the subunit composition and increases the current duration of synaptic NMDA receptors (NMDARs), no such effect is observed at synapses between layer 4 and layer 2/3 pyramidal neurons in adult mice. Surprisingly, visual deprivation in adult mice enhances the temporal summation of NMDAR-mediated currents induced by bursts of high-frequency stimulation. The enhanced temporal summation of NMDAR-mediated currents in deprived cortex could not be explained by a reduction in the rate of synaptic depression, because our data indicate that late-onset visual deprivation actually increases the rate of synaptic depression. Biochemical and electrophysiological evidence instead suggest that the enhanced temporal summation in adult mice could be accounted for by a change in the molecular composition of NMDARs at perisynaptic/extrasynaptic sites. Our data demonstrate that the experience-dependent modifications observed in the adult visual cortex are different from those observed during development. These differences may help to explain the unique consequences of sensory deprivation on plasticity in the developing versus mature cortex. PMID- 16354929 TI - Distinct subunits in heteromeric kainate receptors mediate ionotropic and metabotropic function at hippocampal mossy fiber synapses. AB - Heteromeric kainate receptors (KARs) containing both glutamate receptor 6 (GluR6) and KA2 subunits are involved in KAR-mediated EPSCs at mossy fiber synapses in CA3 pyramidal cells. We report that endogenous glutamate, by activating KARs, reversibly inhibits the slow Ca2+-activated K+ current I(sAHP) and increases neuronal excitability through a G-protein-coupled mechanism. Using KAR knockout mice, we show that KA2 is essential for the inhibition of I(sAHP) in CA3 pyramidal cells by low nanomolar concentrations of kainate, in addition to GluR6. In GluR6(-/-) mice, both ionotropic synaptic transmission and inhibition of I(sAHP) by endogenous glutamate released from mossy fibers was lost. In contrast, inhibition of I(sAHP) was absent in KA2(-/-) mice despite the preservation of KAR mediated EPSCs. These data indicate that the metabotropic action of KARs did not rely on the activation of a KAR-mediated inward current. Biochemical analysis of knock-out mice revealed that KA2 was required for the interaction of KARs with Galpha(q/11)-proteins known to be involved in I(sAHP) modulation. Finally, the ionotropic and metabotropic actions of KARs at mossy fiber synapses were differentially sensitive to the competitive glutamate receptor ligands kainate (5 nM) and kynurenate (1 mM). We propose a model in which KARs could operate in two modes at mossy fiber synapses: through a direct ionotropic action of GluR6, and through an indirect G-protein-coupled mechanism requiring the binding of glutamate to KA2. PMID- 16354928 TI - BACE1, a major determinant of selective vulnerability of the brain to amyloid beta amyloidogenesis, is essential for cognitive, emotional, and synaptic functions. AB - A transmembrane aspartyl protease termed beta-site APP cleavage enzyme 1 (BACE1) that cleaves the amyloid-beta precursor protein (APP), which is abundant in neurons, is required for the generation of amyloid-beta (Abeta) peptides implicated in the pathogenesis of Alzheimer's disease (AD). We now demonstrate that BACE1, enriched in neurons of the CNS, is a major determinant that predisposes the brain to Abeta amyloidogenesis. The physiologically high levels of BACE1 activity coupled with low levels of BACE2 and alpha-secretase anti amyloidogenic activities in neurons is a major contributor to the accumulation of Abeta in the CNS, whereas other organs are spared. Significantly, deletion of BACE1 in APPswe;PS1DeltaE9 mice prevents both Abeta deposition and age-associated cognitive abnormalities that occur in this model of Abeta amyloidosis. Moreover, Abeta deposits are sensitive to BACE1 dosage and can be efficiently cleared from the CNS when BACE1 is silenced. However, BACE1 null mice manifest alterations in hippocampal synaptic plasticity as well as in performance on tests of cognition and emotion. Importantly, memory deficits but not emotional alterations in BACE1( /-) mice are prevented by coexpressing APPswe;PS1DeltaE9 transgenes, indicating that other potential substrates of BACE1 may affect neural circuits related to emotion. Our results establish BACE1 and APP processing pathways as critical for cognitive, emotional, and synaptic functions, and future studies should be alert to potential mechanism-based side effects that may occur with BACE1 inhibitors designed to ameliorate Abeta amyloidosis in AD. PMID- 16354930 TI - Distinct mechanisms for neurotrophin-3-induced acute and long-term synaptic potentiation. AB - Although neurotrophins elicit both acute and long-term effects, it is unclear whether the two modes of action are mediated by the same or different mechanisms. Using neuromuscular junction (NMJ) as a model system, we identified three characteristic features required for long-term, but not acute, forms of synaptic modulation by neurotrophin-3 (NT-3): endocytosis of NT-3-receptor complex, activation of the PI3 kinase substrate Akt, and new protein synthesis. Long-term effects were eliminated when NT-3 was conjugated to a bead that was too large to be endocytosed or when dominant-negative dynamin was expressed in presynaptic neurons. Presynaptic inhibition of Akt also selectively prevented NT-3-mediated long-term effects. Blockade of protein translation by the mammalian target of rapamycin inhibitor rapamycin prevented the long-term structural and functional changes at the NMJ, without affecting the acute potentiation of synaptic transmission by NT-3. These results reveal fundamental differences between acute and long-term modulation by neurotrophins. PMID- 16354931 TI - Trial-by-trial coupling of concurrent electroencephalogram and functional magnetic resonance imaging identifies the dynamics of performance monitoring. AB - Goal-directed behavior requires the continuous monitoring and dynamic adjustment of ongoing actions. Here, we report a direct coupling between the event-related electroencephalogram (EEG), functional magnetic resonance imaging (fMRI), and behavioral measures of performance monitoring in humans. By applying independent component analysis to EEG signals recorded simultaneously with fMRI, we found the single-trial error-related negativity of the EEG to be systematically related to behavior in the subsequent trial, thereby reflecting immediate behavioral adjustments of a cognitive performance monitoring system. Moreover, this trial-by trial EEG measure of performance monitoring predicted the fMRI activity in the rostral cingulate zone, a brain region thought to play a key role in processing of response errors. We conclude that investigations of the dynamic coupling between EEG and fMRI provide a powerful approach for the study of higher order brain functions. PMID- 16354932 TI - Spinal cord-transected mice learn to step in response to quipazine treatment and robotic training. AB - In the present study, concurrent treatment with robotic step training and a serotonin agonist, quipazine, generated significant recovery of locomotor function in complete spinal cord-transected mice (T7-T9) that otherwise could not step. The extent of recovery achieved when these treatments were combined exceeded that obtained when either treatment was applied independently. We quantitatively analyzed the stepping characteristics of spinal mice after alternatively administering no training, manual training, robotic training, quipazine treatment, or a combination of robotic training with quipazine treatment, to examine the mechanisms by which training and quipazine treatment promote functional recovery. Using fast Fourier transform and principal components analysis, significant improvements in the step rhythm, step shape consistency, and number of weight-bearing steps were observed in robotically trained compared with manually trained or nontrained mice. In contrast, manual training had no effect on stepping performance, yielding no improvement compared with nontrained mice. Daily bolus quipazine treatment acutely improved the step shape consistency and number of steps executed by both robotically trained and nontrained mice, but these improvements did not persist after quipazine was withdrawn. At the dosage used (0.5 mg/kg body weight), quipazine appeared to facilitate, rather than directly generate, stepping, by enabling the spinal cord neural circuitry to process specific patterns of sensory information associated with weight-bearing stepping. Via this mechanism, quipazine treatment enhanced kinematically appropriate robotic training. When administered intermittently during an extended period of robotic training, quipazine revealed training induced stepping improvements that were masked in the absence of the pharmacological treatment. PMID- 16354933 TI - Virally mediated increased neurotensin 1 receptor in the nucleus accumbens decreases behavioral effects of mesolimbic system activation. AB - Dopamine receptor agonist and NMDA receptor antagonist activation of the mesolimbic dopamine system increases locomotion and disrupts prepulse inhibition of the acoustic startle response (PPI), paradigms frequently used to study both the pharmacology of antipsychotic drugs and drugs of abuse. In rats, virally mediated overexpression of the neurotensin 1 (NT1) receptor in the nucleus accumbens antagonized d-amphetamine- and dizocilpine-induced PPI disruption, hyperlocomotion, and D-amphetamine-induced rearing. The NT receptor antagonist SR 142948A [2-[[5-(2,6-dimethoxyphenyl)-1-(4-N-(3-dimethylaminopropyl)-N methylcarbamoyl)-2-isopropylphenyl)-1H-pyrazole-3-carbonyl]amino] adamantane-2 carboxylic acid, hydrochloride] blocked inhibition of dizocilpine-induced hyperlocomotion mediated by overexpression of the NT1 receptor. Together, these results suggest that increased nucleus accumbens NT neurotransmission, via the NT1 receptor, can decrease the effects of activation of the mesolimbic dopamine system and disruption of the glutamatergic input from limbic cortices, resembling the action of the atypical antipsychotic drug clozapine. In contrast to clozapine, virally mediated overexpression of the NT1 receptor in the nucleus accumbens had prolonged protective effects (up to 4 weeks after viral injection) without perturbing baseline PPI and locomotor behaviors. These data further confirm the NT1 receptor as the receptor mediating the antistimulant- and antipsychotic-like properties of NT and provide rationale for the development of NT1 receptor agonists as novel antipsychotic drugs. In addition, the NT1 receptor vector might be a valuable tool for understanding the mechanism of action of antipsychotic drugs and drugs of abuse and may have potential therapeutic applications. PMID- 16354934 TI - Specificity in the projections of prefrontal and insular cortex to ventral striatopallidum and the extended amygdala. AB - The basal forebrain functional-anatomical macrosystems, ventral striatopallidum, and extended amygdala are innervated by substantially coextensive distributions of neurons in the prefrontal and insular cortex. This suggests two alternative organizational schemes: convergent, in which a given cortical area projects exclusively to only one of these macrosystems and divergent, in which a given cortical area innervates both forebrain macrosystems. To examine the underlying organization and possibly discriminate between these alternatives, rats were injected with two retrograde tracers in different parts of ventral striatopallidum or extended amygdala (homotypic injection pairs) or with one tracer in each macrosystem (heterotypic). The prefrontal and insular cortex was evaluated microscopically for overlap of retrograde labeling and double labeling of neurons. Homotypic injection pairs in the ventral striatum and extended amygdala produced extensive overlap of retrogradely labeled neurons and significant double labeling, suggesting that cortical projections spread broadly within macrosystems. In contrast, heterotypic injection pairs produced significant overlap of retrograde labeling but negligible double labeling, indicating that ventral striatopallidum and extended amygdala receive inputs from separate sets of prefronto- and insular cortical neurons. The caudomedial shell of the nucleus accumbens, a supposed "transition" zone between striatopallidum and extended amygdala, had extended amygdala-like afferents but produced few double-labeled neurons and these only when paired with ventral striatopallidum. The data suggest that a modular organization of the basal forebrain, with postulated independent information processing by the ventral striatopallidal and extended amygdala macrosystems, is reflected in a corresponding segregation of output neurons in the prefrontal and insular cortices. PMID- 16354935 TI - Inflammatory pain upregulates spinal inhibition via endogenous neurosteroid production. AB - Inhibitory synaptic transmission in the dorsal horn (DH) of the spinal cord plays an important role in the modulation of nociceptive messages because pharmacological blockade of spinal GABAA receptors leads to thermal and mechanical pain symptoms. Here, we show that during the development of thermal hyperalgesia and mechanical allodynia associated with inflammatory pain, synaptic inhibition mediated by GABAA receptors in lamina II of the DH was in fact markedly increased. This phenomenon was accompanied by an upregulation of the endogenous production of 5alpha-reduced neurosteroids, which, at the spinal level, led to a prolongation of GABAA receptor-mediated synaptic currents and to the appearance of a mixed GABA/glycine cotransmission. This increased inhibition was correlated with a selective limitation of the inflammation-induced thermal hyperalgesia, whereas mechanical allodynia remained unaffected. Our results show that peripheral inflammation activates an endogenous neurosteroid-based antinociceptive control, which discriminates between thermal and mechanical hyperalgesia. PMID- 16354936 TI - Hedonic hot spot in nucleus accumbens shell: where do mu-opioids cause increased hedonic impact of sweetness? AB - Mu-opioid systems in the medial shell of the nucleus accumbens contribute to hedonic impact ("liking") for sweetness, food, and drug rewards. But does the entire medial shell generate reward hedonic impact? Or is there a specific localized site for opioid enhancement of hedonic "liking" in the medial shell? And how does enhanced taste hedonic impact relate to opioid-stimulated increases in food intake? Here, we used a functional mapping procedure based on microinjection Fos plumes to localize opioid substrates in the medial shell of the nucleus accumbens that cause enhanced "liking" reactions to sweet pleasure and that stimulate food intake. We mapped changes in affective orofacial reactions of "liking"/"disliking" elicited by sucrose or quinine tastes after D Ala2-N-Me-Phe4-Glycol5-enkephalin (DAMGO) microinjections in rats and compared hedonic increases to food intake stimulated at the same sites. Our maps indicate that opioid-induced increases in sucrose hedonic impact are generated by a localized cubic millimeter site in a rostrodorsal region of the medial shell. In contrast, all regions of the medial shell generated DAMGO-induced robust increases in eating behavior and food intake. Thus, our results identify a locus for opioid amplification of hedonic impact and reveal a distinction between opioid mechanisms of food intake and hedonic impact. Opioid circuits for stimulating food intake are widely distributed, whereas hedonic "liking" circuits are more tightly localized in the rostromedial shell of the nucleus accumbens. PMID- 16354937 TI - A genetic screen for mutations that affect cranial nerve development in the mouse. AB - Cranial motor and sensory nerves arise stereotypically in the embryonic hindbrain, act as sensitive indicators of general and region-specific neuronal development, and are directly or indirectly affected in many human disorders, particularly craniofacial syndromes. The molecular genetic hierarchies that regulate cranial nerve development are mostly unknown. Here, we describe the first mouse genetic screen that has used direct immunohistochemical visualization methods to systematically identify genetic loci required for cranial nerve development. After screening 40 pedigrees, we recovered seven new neurodevelopmental mutations. Two mutations model human genetic syndromes. Mutation 7-1 causes facial nerve anomalies and a reduced lower jaw, and is located in a region of conserved synteny with an interval associated with the micrognathia and mental retardation of human cri-du-chat syndrome. Mutation 22-1 is in the Pax3 gene and, thus, models human Waardenburg syndrome. Three mutations cause global axon guidance deficits: one interferes with initial motor axon extension from the neural tube, another causes overall axon defasciculation, and the third affects general choice point selection. Another two mutations affect the oculomotor nerve specifically. Oculomotor nerve development, which is disrupted by six mutations, appears particularly sensitive to genetic perturbations. Phenotypic comparisons of these mutants identifies a "transition zone" that oculomotor axons enter after initial outgrowth and in which new factors govern additional progress. The number of interesting neurodevelopmental mutants revealed by this small-scale screen underscores the promise of similar focused genetic screens to contribute significantly to our understanding of cranial nerve development and human craniofacial syndromes. PMID- 16354938 TI - Brief report: the relationship between chronic illness and identification and management of psychosocial problems in pediatric primary care. AB - OBJECTIVE: To compare identification, management, and barriers to treatment for psychosocial concerns in children with and without a chronic illness. METHODS: Using data from the Child Behavior Study (CBS), provider rates of identification, treatment, and reports of barriers to treatment were compared in children with and without a chronic illness. Of the 21,065 children ranging in age from 4 to 15 years, 808 children were identified with a chronic illness and were matched on eight demographic variables with 763 healthy children. RESULTS: Providers identified psychosocial concerns in significantly more children with a chronic illness (36.8%) than healthy children (20.2%). Among children with psychosocial concerns, rates of identification, treatment, and barriers to treatment did not differ across the two groups. CONCLUSIONS: Chronic illness did not present more barriers to the management of psychosocial concerns. Increased rates of identification and treatment of psychosocial concerns require attention to general barriers to treatment and screening. PMID- 16354939 TI - Profile of anemia in chronic renal failure patients: comparison between predialyzed and dialyzed patients at the Division of Nephrology, Department of Internal Medicine, Sanglah Hospital, Denpasar, Bali, Indonesia. AB - AIM: To explore the profile of anemia in pradialytic and dialytic CRF patients at the Division of Nephrology, Department of Internal Medicine, Sanglah Hospital, Denpasar, from January to June 2000. METHODS: There were 26 chronic dialyzed patients and 26 pradialyzed patients. Technicon H-1 was used to examine peripheral blood count; blood urea nitrogen and serum creatinine were examined using standard technique. MEIA (microparticle enzyme immunoassay) was used for serum folic acid and serum B12 level. RESULTS: Of 52 CRF patients, hemoglobin (Hb) levels ranged from 4.6 g/dl to 15.1 g/dl with a mean of 9.3 +/- 2.7 g/dl. Hematocrit levels ranged from 15.3% to 49.3%, with a mean of 29.8 +/- 7.9%. There was a significant difference in the hemoglobin and hematocrit levels of chronic dialyzed and pradialyzed patients. The prevalence of anemia (according to the WHO's criteria) among CRF patients was 84.5% (45/52), with the prevalence in chronic dialyzed patients being 100% and 73.1% in pradialyzed patients. If a hemoglobin level of less than 10 g/dl is used as a cut off point, the prevalence of anemia in dialyzed patients was 96.2% and 30.8% among pradialyzed patients. The severity of anemia among 26 dialyzed patients was: severe in 2 cases (8%), moderate in 16 cases (64%) and mild in 7 cases (28%), while in pradialytic patients the severity of anemia was: moderate in 4 patients (50%) and mild in 4 (50%). The morphology of 33 anemic patients was normocrhomic normocytic in 26 (78.8%) cases, slightly macrocytic in 7 (21.2%) cases, and no hypochromic anemia was found. Two anemic patients were associated with low serum folic acid (1 patient with macrocytic anemia and the other case with normochromic normocytic anemia). The serum B12 level was found to be normal in all cases. There was no significant correlation between the severity of anemia and serum creatinine levels or duration of hemodialysis. CONCLUSION: Anemia is a hallmark for CRF patients with a high prevalence of anemia and moderate degree of anemia. Most of anemic cases were normochromic normocytic. Loss of renal mass could be the principle mechanism. In a small proportion of patients, folic acid deficiency may be the cause of anemia. In this study, iron deficiency anemia was not likely an important factor since no hypochromic anemia was found. PMID- 16354940 TI - Microalbuminuria prevalence study in hypertensive patients with type 2 diabetes in Indonesia. AB - AIM: To assess the prevalence of macroalbuminuria and microalbuminuria in hypertensive patients with type 2 diabetes in 10 Asian countries. METHODS: This cross-sectional clinic-based epidemiological study is a subanalysis of data collected from patients attending three medical centres in Indonesia from May 2002 to October 2002. A total of 207 patients were enrolled, of which 177 patients constituted the per protocol population (patients with bacteriuria and haematuria were excluded). RESULTS: Overall, the prevalence of diabetic kidney disease was high, with macroalbuminuria comprising 44.7% (41.2-48.1;95% confidence interval) and microalbuminuria comprising 33.0% (29.7-36.3; 95% confidence interval). While the majority (91.53%) of patients were receiving treatment for hypertension, only 6.21% of the patients had systolic/diastolic blood pressures below the 130/85 mmHg target. CONCLUSION: The prevalence of microalbuminuria and macroalbuminuria was high in hypertensive patients with type 2 diabetes in Indonesia, which is indicative of an impending pandemic of diabetic cardiovascular and renal diseases in the region. PMID- 16354941 TI - The role of persistent anticardiolipin antibody as risk factor of ischemic stroke. AB - AIM: To determine the role of persistent ACA and hyperviscosity as risk factor of ischemic stroke. METHODS: A study was conducted on 76 subjects whose age 40 to 70 years. Subjects consisted of 38 patients of post ischemic stroke and 38 controls with diagnosis other than stroke. Fresh blood samples were taken and mixed with EDTA for viscosity examination and serum for ACA IgM and IgG examination. The laboratory examination for persistent ACA IgM and IgG used ELISA method, while viscosity analysis was using viscometer. Statistic analysis used chi-square and multivariate analysis with logistic regression. RESULTS: In this study we found persistent ACA IgG in 25% of case group , and 2.63% in control group. Multivariate analysis showed persistent ACA IgG as risk factor for ischemic stroke with p < 0.05 and OR 14.11 (CI 95%:1.64;121.11). We found persistent ACA IgM in 2.78% of case group and 5.26% in control group. High blood viscosity was found in 15.79% case group and 10.53% in a control group. Statistical analysis showed no significant difference of viscosity (p = 0.740) and persistent ACA IgM (p = 1.000) between case and control group. CONCLUSION: study showed that persistent ACA IgG in stroke ischemic was higher than in control subjects. Blood viscosity examination and persistent ACA IgM did not show significant difference. While persistent ACA IgG with OR 14.11 (CI: 1.64; 121.11) was the risk factor for ischemic stroke. Blood viscosity and persistent ACA IgM were not risk factors for ischemic stroke. PMID- 16354942 TI - Overview of Smith-Magenis syndrome. AB - Increased awareness of the phenotype associated with SMS permits early diagnosis and optimal management and intervention. From a management standpoint, the unique constellation of behavioral dysfunction and sleep disturbances has a significant impact on the parents, siblings and relatives of individuals with SMS as well as the professionals involved in their care. Support groups for families of persons with SMS like PRISMS in the US [http://www.prisms.org], provide families with up to-date information, as well as emotional and peer support, enabling them to better care for their child. PMID- 16354943 TI - Cell death as a possible mechanism for tissue limited mosaicism in Pallister Killian syndrome. AB - Pallister-Killian syndrome is a chromosomal mosaic syndrome with a normal and an isochromosome 12p cell line, the latter rarely seen in peripheral blood. The isochromosome 12p cell line decreases with serial passages of fibroblasts in vitro and with age of patient in vivo. To evaluate cell death as a possible mechanism for loss of the abnormal cell line, amniocytes from a fetus with Pallister-Killian syndrome were identified as normal or aneuploid using a chromosome 12 alpha-satellite DNA probe by fluorescent in situ hybridization (FISH) and then subsequently stained with Annexin V, which stains the cytoplasm of cells that are dying. Although not conclusive, our preliminary results suggest that the abnormal cell line is going through apoptosis or necrosis at a higher rate than normal cells. Cell death may be a possible mechanism for decrease of the aneuploid cell line in patients with Pallister-Killian syndrome. PMID- 16354945 TI - Environmental factors influencing allergy and asthma. AB - Despite the strong and consistent association between immediate hypersensitivity, allergy, asthma and exposure, there is still controversy about the role inhaled allergen plays and about the timing of events related to sensitization. However, IgE antibodies are essential for the asthmatic response and the symptoms are much more closely associated with IgE antibodies to perennial than to seasonal allergens. Although there have been many changes in our environment, none of these alone explains the consistency or the scale of the rise in asthma over the 40 years between 1960 and 2000. PMID- 16354946 TI - Should asthma management include sojourns at high altitude? AB - Sojourns in the high mountains have been recommended by specialists for patients with asthma since many decades. An inquiry among physicians of the 'Davoser Arzteverein' revealed as early as 1906 that 133/143 patients with bronchial asthma had no or only few asthma attacks during their stay in Davos, and that 81% had a persistent improvement of their disease. These early observations about effects of the alpine climate were, of course, reported at a time, when the spectrum of pharmacotherapy was very limited. However, these observations were consistent and were therefore regarded as proof for the therapeutic value of sojourns under alpine conditions in bronchial asthma. In recent years, however, the indication for asthma treatment in high mountains is increasingly questioned, in particular by health insurance systems. Therefore it is the aim of this contribution to summarize the available data about the effects of a stay of asthmatic patients at 1,500-1,800 m above sea level. It is concluded that the available evidence suggests a significant beneficial effect of high altitude in bronchial asthma, in particular in steroid-dependent patients. PMID- 16354947 TI - The immunological basis of the hygiene hypothesis. AB - The dramatic increase of allergic disorders in the last decades made their study an imperious demand. The increasing incidence of the development of allergic disorders seems to be associated with the modern westernized lifestyle, but causal reasons and the underlying mechanisms are far from being completely understood. Evidences suggest that priming of the immune responses against allergens happens already in utero. In addition, early life events are essential in shaping the immune answer towards the Th1- or Th2-profile, associated with a nonallergic or allergic phenotype, respectively. The hygiene hypothesis suggests that an early life environment rich in normal microbial flora primes the immune system in the Th1 direction towards clinical balance while a 'sterile' environment rather promotes the development of pathological immune phenotypes. In this review we collect epidemiological evidence for this concept. The data suggest an association between environment, lifestyle and the development of allergic diseases. This is the basis for the development of new hypotheses regarding the underlying pathomechanisms. The current view of cellular and molecular mechanisms underlying these phenomena includes fine-balancing between innate immune mechanisms and Th1, Th2 and regulatory T cells. These novel immunoregulatory events may explain the hygiene hypothesis by an interaction of environmental factors with innate immune mechanisms and various subtypes of T cell responses. PMID- 16354948 TI - Molecules involved in the regulation of eosinophil apoptosis. AB - Apoptosis is the most common form of physiological cell death and a necessary process to maintain cell numbers in multicellular organisms. Eosinophils are constantly produced in the bone marrow and the same numbers die, under normal circumstances, within a relatively short time period. In many eosinophilic inflammatory diseases, reduced eosinophil apoptosis has been described. This mechanism may contribute to increased eosinophil numbers, a phenomenon called eosinophilia. Overexpression of interleukin-5 appears to be crucial for delaying eosinophil apoptosis in many allergic disorders. Survival factor withdrawal leads to the induction of apoptosis. Besides survival cytokines, eosinophil apoptosis is also regulated by death factors. Recent observations suggest a role for mitochondria in conducting eosinophil apoptosis, although the mechanisms that trigger mitochondria to release proapoptotic factors remain less clear. Drugs that specifically induce eosinophil apoptosis might be useful for triggering the resolution of unwanted eosinophilic inflammatory responses. PMID- 16354949 TI - The role of T lymphocytes in asthma. AB - There is now overwhelming evidence to support a major role for T cells in asthma, in particular the involvement of T helper type 2 (Th2) cells in atopic allergic asthma as well as nonatopic and occupational asthma. There may also be a minor contribution from T cytotoxic type 2 CD8+T cells. Several Th2 cytokines have potential to modulate airway inflammation, in particular interleukin-13 which induces airway hyperresponsiveness independently of IgE and eosinophilia in animal models. The identification of transcription factors controlling Th1, Th2 and T-regulatory cell (T(Reg)) development further support the Th2 hypothesis since GATA3 is overexpressed and T-bet underexpressed in the asthmatic airway and Foxp3 is induced in asthma by corticosteroid treatment. Specific T-cell-directed immunotherapy may allow induction/modulation of T-cell responses, and elucidation of the mechanisms of T(Regs) may allow further optimization of immunotherapy. Recent advances in the understanding of dendritic cell function in directing T cell responses may uncover further therapeutic targets. Efficacy of cyclosporin and anti-CD4 treatment in chronic severe asthma argues for continued T-cell involvement, but whether remodeling contributes to pathology inaccessible to antiinflammatory treatment or T-cell immunotherapy remains an important question. PMID- 16354950 TI - Allergic manifestations of skin diseases--atopic dermatitis. AB - Atopic dermatitis (AD) is a chronic inflammatory skin disease which often becomes manifest in early infancy and is characterized by itchy eczematous lesions with characteristic localization. The cellular infiltrate of allergic eczematous skin diseases (i.e. AD, allergic contact dermatitis) is mainly composed of mononuclear cells. Whereas allergic contact dermatitis is always triggered by allergen specific T cells, a number of allergic and nonallergic trigger factors appear to be relevant in AD. This article discusses data coming from immunological studies focusing on T-cell responses in AD. The concept of a switch from a T helper type 1 (Th1) to a Th2 cytokine profile in lesional skin of AD is well accepted. Besides CD4+ T lymphocytes, CD8+ cells are likely to play an important role in the pathogenesis of AD. Recent studies point to the induction of apoptosis in keratinocytes by interferon-gamma derived from skin-homing T cells as a further important mechanism for the induction and maintenance of the eczema. Recent clinical studies have confirmed the major role of food allergy and infectious microorganisms as trigger factors of AD. New therapeutic strategies for AD include topical calcineurin inhibitors which were introduced as a new therapeutic principle at the beginning of this decade. PMID- 16354951 TI - Skin-homing T cells in cutaneous allergic inflammation. AB - The involvement of circulating cutaneous lymphocyte-associated (CLA)+ T cells in skin allergy was initially described in atopic and contact dermatitis in a series of experiments performed in Davos, at the Swiss Institute of Allergy and Asthma Research. Since then, numerous studies have confirmed and extended those initial findings. Both basic and clinical research data obtained on this subset of memory T cells support a relevant role of CLA+ T cells in the regional cutaneous immune system. The spectrum of cutaneous diseases where CLA+ T cells have been shown to play a relevant role has expanded far beyond the field of allergy, highlighting the relevance of this memory T-cell subpopulation in cutaneous diseases. The goal of this chapter is to review known features of those cells in atopic dermatitis, contact dermatitis and nonimmediate cutaneous allergic reactions to drugs. PMID- 16354952 TI - The role of sensitization to Malassezia sympodialis in atopic eczema. AB - Atopic eczema (AE) is a chronic relapsing, highly pruritic inflammation of the skin with a worldwide prevalence of 10-20% in children and of 1-3% in adults. Malassezia sympodialis has been reported as the most frequent skin-colonizing yeast in both AE patients and healthy individuals. Approximately 50% of the AE patients show immediate-type skin reactions or have specific serum IgE against M. sympodialis. Sensitization to the yeast occurs almost exclusively in AE patients. The main cause for this specific sensitization may be the disrupted skin barrier facilitating allergen uptake. So far thirteen allergens of Malassezia have been cloned produced, characterized and partly studied in vitro and in vivo. Phylogenetically conserved allergen structures, such as manganese superoxide dismutase, may play a role as cross-reactive allergens in a subset of AE patients as a result of molecular mimicry and cross-reactivity with structurally related human proteins and might contribute to the perpetuation of the inflammatory skin reactions. The use of recombinant Malassezia allergens will contribute to elucidate the pathways of sensitization occurring in AE, the underlying immunological mechanisms governing IgE- and T-cell-mediated responses and may provide new therapeutic options to alleviate Malassezia-related symptoms in AE. PMID- 16354953 TI - Allergic conjunctivitis: the forgotten disease. AB - The eye is largely exposed to foreign substances, and ocular tissues have a complete array of immune cells to interact with offending antigens. In particular, the external eye represents an ideal site for immediate hypersensitivity reactions because of the high number of mast cells in the eyelids and bulbar conjunctiva, as well as for the potential local synthesis of IgE. In fact, the involvement of the eye was part of the original description of hay fever by Charles Blackley in 1873. In a large epidemiological survey of allergic patients we reported since 1987, 40% had symptoms possibly related to allergic conjunctival disease. However, the participation of the conjunctiva to the multiorgan picture of allergy is largely underestimated. Allergic eye diseases have long been referred to the classical Type I hypersensitivity mechanism according to Coombs and Gell. Recently, however, the tremendous progress in understanding allergic mechanisms and inflammation has brought new insights into the pathophysiology of several allergic diseases, including those of the eye. Accordingly, several systematic descriptions of allergic eye diseases should be revisited. In particular, the classification of the various forms of a 'red eye' of allergic origin appears inadequate to answer the progress achieved in their multifactorial pathogenesis. PMID- 16354954 TI - Fungal allergies: a yet unsolved problem. AB - Airborne fungal spores have been implicated as causative factors in respiratory allergy, particularly asthma. However, the prevalence of fungal sensitization is not known mainly due to the lack of standardized fungal extracts and to the overwhelming number of fungal species able to elicit IgE-mediated reactions. Recent work based on high-throughput cloning of fungal allergens revealed that fungi are able to produce extremely complex repertoires of species-specific and cross-reactive allergens. There is evidence that fungal sensitization also contributes to auto-reactivity against self-antigens due to shared epitopes with homologous fungal allergens. Detailed studies at structural and immunological level indicate molecular mimicry as a basic mechanism involved in perpetuation of severe chronic allergic diseases. The real challenge at present is not related to cloning or production of a large number of different fungal allergens but rather to the assessment of the clinical relevance of each single structure. To date, substitution of complex extracts presently used in the diagnosis of fungal allergy by single, perfectly standardized components seems feasible in contrast to specific immunotherapy which is still not developed. Recombinant fungal allergens might create new perspectives in diagnosis and therapy of fungal allergy. PMID- 16354955 TI - Structural features of allergenic molecules. AB - In this paper the relation between protein allergenicity (the capacity to induce IgE antibody production or the capacity to activate mast cells sensitized with IgE antibodies induced by a cross-reactive allergen) and protein structure is discussed. While cross-reactivity is to a large degree predictable from primary sequence comparisons, the IgE-inducing capacity is mostly determined by factors other than the primary sequence. Two routes to IgE are discussed: (1) the atopic route (used by allergens from pollen and mites) in which a direct switch from mu to epsilon is common and (2) the 'modified Th2' route (used by allergens from pets) in which the class switch to IgE is often preceded by a switch to IgG4. According to this working hypothesis, the choice between these two routes is determined at the level of the germinal center activity. PMID- 16354956 TI - Regulation of human T helper cell differentiation by antigen-presenting cells: the bee venom phospholipase A2 model. AB - Whereas some individuals develop immunity to bee sting and mount protective IgG4- mediated antibody responses to bee venom phospholipase A2 (PLA), others produce large amounts of PLA-specific IgE antibodies and become allergic to this, otherwise, innocuous antigen. PLA-specific IgE responses are the result of imbalanced T helper (Th)2-cell differentiation. There are multiple mechanisms driving the differentiation of naive CD4+ T cells into Th1- or Th2-cell phenotypes. Most of them are linked to the conditions occurring during initial or repeated encounters with the allergen, in the context of an antigen-presenting cell (APC). The different types of APC and their availability to display particular cytokine production profiles, pattern recognition receptors, costimulatory molecules and specific HLA haplotypes are key determinants for human Th1- and Th2-cell polarization. PMID- 16354957 TI - T regulatory cells in allergy. AB - Activation-induced cell death, anergy and/or immune response modulation by T regulatory cells (T(Reg)) are essential mechanisms of peripheral T-cell tolerance. There is growing evidence that anergy, tolerance and active suppression are not entirely distinct, but rather, represent linked mechanisms possibly involving the same cells and multiple suppressor mechanisms. Skewing of allergen-specific effector T cells to T(Reg) cells appears as a crucial event in the control of healthy immune response to allergens and successful allergen specific immunotherapy. The T(Reg) cell response is characterized by abolished allergen-induced specific T-cell proliferation and suppressed T helper 1 (Th1)- and Th2-type cytokine secretion. The increased levels of interleukin-10 (IL-10) and transforming growth factor-Beta (TGF-Beta) that are produced by T(Reg) cells, potently suppress IgE production, while simultaneously increasing production of noninflammatory isotypes IgG4 and IgA, respectively. In addition, T(Reg) cells directly or indirectly suppress effector cells of allergic inflammation such as mast cells, basophils, and eosinophils. In conclusion, peripheral tolerance to allergens is controlled by multiple active suppression mechanisms. It is associated with regulation of antibody isotypes and effector cells to the direction of a healthy immune response and opens a window for novel therapies of allergic diseases. PMID- 16354958 TI - The role of histamine in regulation of immune responses. AB - Histamine is not only the major mediator of the acute inflammatory and immediate hypersensitivity responses, but has also been demonstrated to affect chronic inflammation and regulate several essential events in the immune response. It can influence numerous functions of the cells involved in the regulation of immune response and hematopoiesis including macrophages, dendritic cells, T lymphocytes, B lymphocytes and endothelial cells. These cells express histamine receptors and also secrete histamine, which can selectively recruit the major effector cells into tissue sites and affect their maturation, activation, polarization, and effector functions leading to chronic inflammation. Histamine regulates antigen specific Th1 and Th2 cells, as well as related antibody isotype responses. Histamine acting through its receptor (HR) type 2, positively interferes with the peripheral antigen tolerance induced by T regulatory (T(Reg)) cells in several pathways. The diverse effects of histamine on immune regulation are due to differential expression and regulation of 4 histamine receptors and their distinct intracellular signals. In addition, differences in affinities of these receptors are highly decisive on the biological effects of histamine and agents that target histamine receptors. Although substantial evidence has been accumulated about histamine metabolism, receptors, signal transduction, physiological and pathological effects, the complex interrelationship and cross talk by histamine, its receptors and other G-protein coupled receptors remain to be understood. PMID- 16354959 TI - Gene expression profiling in allergy and asthma. AB - DNA array technique was applied for the simultaneous analysis of multiple genes for the characterization of diseases and for cellular analysis. This technology implicated genes in the pathogenesis of allergic disease, which were previously not discussed in this context. This article summarizes results of gene expression profiling of allergic disease and immunologically relevant cells, using DNA array technology. The results of these studies are searchable in public databases and represent exciting tools to study gene expression of diseases on a global base. PMID- 16354960 TI - Mechanisms of allergen-specific immunotherapy. AB - Allergen-specific immunotherapy (SIT) is the only treatment, which leads to a life-long tolerance against allergens due to restoration of normal immunity. The induction of a tolerant state in peripheral T cells represents an essential step in allergen-SIT. Peripheral T-cell tolerance is characterized mainly by suppressed proliferative and cytokine responses against the major allergens and its T-cell recognition sites. It is initiated by autocrine action of IL-10 and/or TGF-Beta, which are increasingly produced by the antigen-specific T Regulatory (T(Reg)) cells. Tolerized T cells can be reactivated to produce either of the distinct Th1 or Th2 cytokine patterns, thus directing allergen-SIT towards successful or unsuccessful treatment. T(Reg) cells directly or indirectly influence effector cells of allergic inflammation, such as mast cells, basophils and eosinophils. In addition, there is accumulating evidence that they may suppress IgE production and induce IgG4 and IgA production against allergens. By the application of the recent knowledge in mechanisms of allergen-SIT, more rational and safer approaches are awaiting in the future for the prevention and cure of allergic diseases. PMID- 16354961 TI - Regulation of the IgE response at the molecular level: impact on the development of systemic anti IgE therapeutic strategies. AB - The classical allergic reaction starts within seconds or minutes after antigen contact and is induced by antibodies produced by a special subset of B lymphocytes. These antibodies belong to the IgE subclass and are responsible for Type I hyper-reactivity reactions. IgE plays a minor role in healthy individuals. In allergic individuals, however, IgE antibodies trigger allergic responses through allergen-mediated cross-linking on effector cells followed by mediator release. The mechanisms inducing a switch to IgE production are not fully understood with the consequence that allergies are mainly treated with antisymptomatic drugs. To develop basic therapies, many questions concerning the very complex regulation of IgE expression have to be understood. Positive and negative regulators influence the synthesis of IgE. Experiments in our laboratory could show that not only regulatory molecules, but also the membrane bound IgE itself controls the quantity and quality of the IgE produced. This fact becomes more and more interesting, because the signals generated by the B-cell receptor may be important targets for interference in allergic patients, in whom the titer and the affinity of the IgE antibodies for the allergen are directly related to disease activity. PMID- 16354962 TI - Gallbladder sludge and acute pancreatitis induced by acute hepatitis A. AB - In this case report, a young woman with gallbladder sludge and acute pancreatitis due to acute hepatitis A (HAV) is presented. She was admitted to our hospital with abnormal hepatic enzymes. Five days prior to her admission, an initial abdominal ultrasound was performed at another hospital and revealed no abnormality, while her serum aspartate aminotransferase (AST) level was at the upper limit of normal (ULN) x 8. A second ultrasound was performed at our hospital and revealed a gallbladder wall thickness (9.3 mm), gallbladder sludge in the gallbladder lumen, pancreatic edema, ascites, and hepatomegaly while AST was at the ULN x 50. Magnetic resonance imaging and magnetic resonance cholangiopancreatography revealed imaging features of an acute stage of pancreatitis and gallbladder wall thickness with coexisting sludge in the gallbladder lumen. HAV infection was diagnosed by the detection of immunoglobulin M against HAV in the serum. The patient underwent two repeated abdominal ultrasound examinations on the 5th (AST was at the ULN x 3) and the 20th days (AST was at the normal) after her discharge, and both revealed normal findings. In our case, we observed reversible changes in the hepatobiliary and pancreatic system which was related to the severity of hepatic necro-inflammation. HAV associated pancreatitis may be due to the formation of biliary sludge during the acute phase of the viral illness, but this association needs further investigation. PMID- 16354963 TI - Inflammatory myofibroblastic tumor of the distal bile duct associated with lymphoplasmacytic sclerosing pancreatitis. Case report and review of the literature. AB - Inflammatory myofibroblastic tumor (IMT) or inflammatory pseudotumor has been described in various organs such as the liver, intestinal tract, spleen, kidney, bladder, lung, peritoneum and heart. However, its appearance in the periampullary region is uncommon and has rarely been reported in the literature. It is characterized histologically by myofibroblastic cell proliferation together with a mixed inflammatory infiltrate that clinically and radiologically mimics a malignant tumor. We report a case of IMT located in the distal common bile duct of a 51-year-old woman. She underwent Whipple resection with the initial diagnosis of cholangiocarcinoma; the pathologic diagnosis of the tumor was IMT of the distal bile duct associated with lymphoplasmacytic sclerosing pancreatitis. Referring to previously reported cases, suspected diagnosis of a malignant tumor made surgical excision the primary choice for symptom relief and in order to obtain a definitive diagnosis. IMT relationship with lymphoplasmacytic sclerosing pancreatitis is discussed. PMID- 16354964 TI - Radiofrequency ablation has a valuable therapeutic role in metastatic VIPoma. AB - BACKGROUND: Vasoactive intestinal peptide-secreting tumours (VIPomas) are rare islet cell tumours of the pancreas that can result in life-threatening biochemical abnormalities. The optimal intervention for metastatic VIPoma remains undecided. This case history documents the clinical role of radiofrequency (RF) ablation in the treatment of metastatic VIPoma. CASE HISTORY: A primary pancreatic VIPoma was diagnosed in a 61-year-old female in 1998 and a distal pancreatectomy and splenectomy were performed. She remained disease-free for 44 months when she presented as an emergency with watery diarrhoea, hypokalaemia, renal failure and an elevated serum VIP level. CT scanning showed a liver metastasis and open RF ablation was performed with complete resolution of symptoms and biochemistry within 48 h. Post-ablation imaging confirmed complete ablation of the metastasis. She remained disease-free until 22 months later when watery diarrhoea resumed and a new hepatic metastasis was seen on CT. Percutaneous RF ablation was performed and follow-up CT scan showed complete ablation of the metastasis. The patient remains disease- and symptom-free 10 months after the second RF ablation. CONCLUSION: This case illustrates that the pronounced clinical and biochemical upset caused by metastatic VIPoma can be resolved safely, quickly and repeatedly by RF ablation. PMID- 16354965 TI - Recurrent pancreatitis in a patient with autosomal-dominant polycystic kidney disease. AB - Autosomal-dominant polycystic kidney disease is an inherited disorder characterized by multiple cysts in kidneys and other organs. A 63-year-old man was evaluated for the etiology of recurrent pancreatitis and chronic renal failure. Multiple cysts of kidneys, liver, and pancreas and pancreas divisum was diagnosed. Pancreatitis should be included in the differential diagnosis of abdominal pain in patients with ADPKD. Pancreas divisum may be a predisposing factor for acute pancreatitis in these patients. PMID- 16354966 TI - Placental lesions in a case of DiGeorge sequence. AB - This work describes some placental alterations found in a partial form of DiGeorge sequence, namely, hypoplasia of a cord artery with internal calcification of an extensive endoluminal thrombosis, and widespread calcification of microthrombi in the arteries of the second and third order villous branches. Hypoplasia of a cord artery is a relatively rare event, and is also associated with malformations of the gastroenteric and cardiovascular system, as sometimes described in the DiGeorge sequence. Interesting placental alterations are reported and their likely physiopathologic basis and pathogenic correlation discussed in order to give a better and more comprehensive picture of the DiGeorge sequence in which the correlated placental alterations are not sufficiently known. PMID- 16354967 TI - Massive feto-maternal hemorrhage: diagnosis by cardiotocography, Doppler ultrasonography and ST waveform analysis of fetal electrocardiography. AB - A 34-year-old healthy gravida 2 para 1 presented after an uncomplicated pregnancy at term with a 2-day history of diminished fetal movements. Fetal anemia was suspected by fetal heart rate monitoring and Doppler estimation of the fetal peak blood flow velocity of the middle cerebral artery. We were also fortunate to register pathological ST waveform changes of the fetal ECG indicating fetal hypoxia. The diagnosis of a massive feto-maternal hemorrhage was confirmed by an extremely high fraction of erythrocytes containing fetal hemoglobin in maternal blood and, after delivery, by placental histology. PMID- 16354968 TI - Fetal magnetic resonance imaging of an intracranial venous thrombosis. Case report. AB - OBJECTIVE: To describe the magnetic resonance imaging findings of an antenatal intracranial venous thrombosis. METHODS: Prenatal ultrasound (US) at 22 weeks' gestational age (GA) in a 27-year-old patient (G2 P1) revealed a nonspecific avascular echogenic mass above the cerebellum evaluated with MRI at 23 weeks' GA. RESULTS: With MRI an enlarged occipital pericerebral space with a layered ovoid mass at the torcular herophili was demonstrated and MRI characteristics suggested a subacute cloth. In follow-up, an important enlargement of the mass was noticed on US examinations. After multidisciplinary consult and parental consent, a prostaglandin induction was performed at 25 weeks' GA. The baby died immediately after birth. Pathology confirmed the diagnosis of a venous thrombosis at the torcular herophili with extension into the superior sagittal sinus. CONCLUSION: In our case, MRI findings were helpful in diagnosing an intracranial fetal venous thrombosis. PMID- 16354969 TI - Prenatal diagnosis of hypochondroplasia: three-dimensional multislice computed tomography findings and molecular analysis. AB - We report the first case of sporadic hypochondroplasia diagnosed in utero by computed tomography (CT) three-dimensional (3D) imaging and molecular analysis at 38 weeks' gestation. Prenatal sonographic examinations performed at 32 and 35 weeks' gestation revealed a rhizomelic shortness of the long bones (femur and humerus) with macrocephaly. Based on these findings, a nonlethal form of skeletal dysplasia was suspected and a multislice CT imaging with 3D reconstruction was performed depicting skeletal abnormalities which suggested hypochondroplasia. The prenatal diagnosis was confirmed by DNA mutation analysis of the fibroblast growth receptor 3 gene. PMID- 16354970 TI - Optimal interval between middle cerebral artery velocity measurements when monitoring pregnancies complicated by red cell alloimmunization. AB - OBJECTIVE: To evaluate the optimal interval between middle cerebral artery (MCA) Doppler measurements when monitoring pregnancies complicated by red cell alloimmunization. METHODS: Thirty-nine fetal blood samplings (FBS) performed on 24 pregnant women with red blood cell alloimmunization followed up using both MCA peak systolic velocity and time-averaged mean velocity measurements on weekly basis. RESULTS: In total, 65.5 and 37.5% of women with moderate or severe fetal anemia had abnormal MCA Doppler values 1 and 2 weeks, respectively, before FBS was performed. CONCLUSIONS: A weekly assessment of women at risk for fetal anemia is optimal in most of the cases even though 35.5% of cases of moderate or severe fetal anemia are expected to have normal Doppler measurements the week before the decision of doing an FBS is made. PMID- 16354971 TI - Size and location of the fetal human ovary. AB - OBJECTIVE: To determine the development and the localization of the ovaries during the fetal period. MATERIAL AND METHODS: One hundred and fifty-four ovaries obtained from 77 human fetuses aged between 9 and 40 weeks of gestation were used in this study. Firstly, the shapes and the positions of the ovaries were established. Second, the localization of the ovaries with respect to linea terminalis, ureters, and the iliac arteries were determined. Finally, the dimensions and the weight of the ovaries were measured. FINDINGS: In the fetal period, the ovaries were most commonly almond shaped and had an oblique orientation. In the 1st trimester the midpoint of the long axis of the fetal ovaries were at the level of linea terminalis. In the 2nd and 3rd trimester and full-term fetuses, it was observed that the ovaries were not in ovarian fossa, suggesting that descensus ovary was in progression during these times. During the intrauterine period, the ovaries were most commonly located anterior to the ureters and over the common iliac artery, only to migrate to its final location between the internal and external iliac arteries towards the end of the 40th week. CONCLUSION: We found that the ovaries did not assume the position of the adults at the end of the fetal period, rather continued its descent after the birth. We believe our findings about the fetal ovaries will be useful in obstetrics, fetal pathology, and forensic pathology. PMID- 16354972 TI - Sonographic cervical length measurement before labor induction in term nulliparous women. AB - OBJECTIVE: The purpose of the study was to determine if transvaginal sonographic measurement of the cervical length is a useful method to predict successful labor induction in nulliparas. METHODS: 137 women who were scheduled for medically indicated induction of labor had a transvaginal sonographic measurement of the cervical length before labor induction. Inclusion criteria were: (1) singleton pregnancy; (2) gestational age between 37-42 weeks; (3) live fetus in cephalic presentation; (4) intact membranes; (5) no vaginal bleeding; (6) no previous history of uterine surgery; (7) nulliparous women, and (8) no allergy or asthma in response to prostaglandins. Induction of labor was performed within 6 h of the ultrasonographic examination, by inserting 2 mg of dinoprostone in the posterior vaginal fornix, repeated if needed every 6 h for up to three doses. When the cervix became favorable and no regular contractions were observed, amniotomy and oxytocin augmentation, starting at 1 mIU/min and increasing 1 mIU every 30 min as necessary, was performed. RESULTS: All women were Caucasians and the mean age was 24.3 years (range 19-37 years). The mean cervical length was 28 mm (range 11-39 mm). The Bishop score was < or =5 in 101 women and >5 in the 36 others. Vaginal delivery occurred in 92 women (67.1%), and the vast majority of them (89 women; 96.7%) gave birth within 24 h of induction. Forty-five women (32.8%) had a cesarean section. The Bishop score was not predictive of the mode of delivery. Thirty-six of 101 women (35.6%) with a Bishop score < or =5 delivered by cesarean section, compared to 9 of 36 women with a Bishop score >5 (25%) (p = NS). Women with a cervical length <27 mm were more likely to deliver vaginally. Using this cutoff value the sensitivity of a successful labor induction was 76% and the specificity was 75.5%. CONCLUSIONS: Transvaginal sonographic measurement of cervical length is a good predictor of a successful labor induction at term in nulliparas. PMID- 16354973 TI - Fetal lung growth in congenital diaphragmatic hernia. AB - BACKGROUND: In spite of significant therapeutic progress, the prognosis of congenital diaphragmatic hernia (CDH) remains pejorative in those forms in which the liver is herniated into the chest. The severity of this malformation relies on the pulmonary hypoplasia due to lung compression by the herniated viscera in the thoracic cavity, particularly the liver. This impaired growth concerns the whole pulmonary tissue, i.e. both the vessels and the alveoli. For the clinician, it is mandatory to know the evolution pattern of the lesions, to define the best time to treat them. AIM AND METHOD: The aim of this work was to study the pulmonary lesions along the gestation in fetuses affected byCDH. This morphological study was carried out on 134 human fetuses aged from 22 to 40 weeks of gestation. Anatomical and histological analysis focused on lung weight, alveolar count and wall thickness of the distal vessels. RESULTS: The results indicate that the pulmonary lesions worsen as the pregnancy continues, particularly beyond 30 weeks of gestation. CONCLUSION: Such an anatomical study should bring to the clinicians useful data to enhance the management of the patients. PMID- 16354974 TI - Evaluation levels of cytokines in amniotic fluid of women with intrauterine infection in the early second trimester. AB - Amniotic fluid was obtained from 180 patients by amniocentesis at 16-22 weeks of gestation and assayed for the levels of interleukin (IL)-6, IL-8, leukocyte elastase (LE), and glucose. Ten of cases had clinical symptoms, such as uterine contraction, genital bleeding, and cervical ripening, and the other 170 were assessed for fetal chromosomal features. Four of the ten cases with uterine contraction developed abortion, while 10 of those screened had findings of fetal chromosomal anomalies, and 7 cases then underwent induced abortion artificially. In the cases of abortion, levels of IL-6, IL-8 and LE were higher than in the samples from the 160 pregnant women without clinical symptoms and a normal karyotype, while glucose in amniotic fluid was lower. Of 6 cases with clinical symptoms, but not developing abortion, 4 developed preterm labor, and in these IL 6 and IL-8 also were significantly elevated, with LE being slight high compared to normal. The results suggest that IL-6, IL-8, LE, and glucose in amniotic fluid at early second trimester can be used as markers of severe infection in the uterus, and with the first two being particularly sensitive. PMID- 16354975 TI - Manifestation of mirror syndrome after fetoscopic laser photocoagulation in severe twin-twin transfusion syndrome. AB - Mirror syndrome is a preeclampsia-like disease first described in a case of severe hydrops fetalis caused by rhesus isoimmunization, later reported in some cases of nonimmunological fetal hydrops. Twin-twin transfusion syndrome (TTTS) is a severe complication associated with monochorionic pregnancies, in particular, severe TTTS with one hydropic fetus leading to a poor prognosis. We report here a case of mirror syndrome that occurred after selective fetoscopic laser photocoagulation in severe TTTS at 24 weeks' gestation. PMID- 16354976 TI - Intravenous immunoglobulins without initial and follow-up cordocentesis in alloimmune fetal and neonatal thrombocytopenia at high risk for intracranial hemorrhage. AB - OBJECTIVE: To report on a less invasive treatment strategy in alloimmune fetal and neonatal thrombocytopenia (FNAIT) at high risk for either in utero or neonatal intracranial hemorrhage (ICH). METHODS: In 7 pregnancies, with a history of ICH in the older sibling, weekly intravenous immunoglobulin (IVIG) therapy to the mother (1 g/kg) without initial cordocentesis was started at a median gestational age of 16 weeks. RESULTS: In 4 pregnancies cordocentesis was avoided. One predelivery cordocentesis with platelet transfusion was performed in 3 further cases. Although none of the cases had a platelet count of >50 x 10(9)/l at cordocentesis, predelivery or birth, no ICHs were observed. The neonatal periods of the infants were uncomplicated. CONCLUSION: IVIG treatment alone might be considered in patients with both severe platelet alloimmunization and an increased risk for morbidity and mortality at cordocentesis. PMID- 16354977 TI - Hemodynamic analysis of cephalothoracopagus by the color doppler method. A comparison to normal fetuses via a longitudinal study. AB - In a case of cephalothoracopagus, the umbilical artery (UA) was observed with color Doppler method, and the findings were compared with the hemodynamics of 46 normal fetuses. The patient was a 25-year-old primigravida who had appeared for routine prenatal visits since her 6th week of pregnancy. At a later time, the patient was examined after an interval of 4 weeks. Although an ultrasonography was also conducted, unfortunately, any findings of cephalothoracopagus were not detected. In the 25th week of gestation, we hospitalized her for marked polyhydramnios (amniotic fluid index: 280 mm), at which time an ultrasound examination revealed cephalothoracopagus. In the UA, the V(max) was 30.3 cm/s (normal fetus at 25-28 weeks: 33.5 +/- 3.9 cm/s). The UA hemodynamics fell below the normal range. At 26 weeks, the UA V(max) was 56.5 cm/sec, a level which significantly exceeded the normal range. The patient underwent a cesarean section at 27 weeks of gestation; the indication was fetal distress. This is caused by the condition in which the fetal heart beats decreases to 90 beats per minute 3 times during a 10-min period as measured on the cardiotocograms. She delivered a 1,392-gram female with an Apgar score of 2 points (respiratory 1 point and heart rate 1 point). The infant was a cephalothoracopagus, with one head, two hearts, four upper limbs, and four lower limbs. The neonate died from circulatory failure 56 min after birth. PMID- 16354978 TI - Triploidy in a growth discordant twin pregnancy after intracytoplasmic sperm injection treatment. Case report. AB - OBJECTIVE: We report on a case of a triploidy in one fetus of a twin pregnancy who presented with the findings of growth discordance. METHODS: Three weeks' difference between fetuses in ultrasonographic measurements in the first trimester was observed in a twin pregnancy following intracytoplasmic sperm injection (ICSI) treatment for male infertility. Hydrocephaly developed in the growth-discordant fetus subsequently. Amniocentesis for both fetuses and selective feticide for the abnormal one was performed at 19 weeks' gestation through a single insertion. Fetal karyotype of the abnormal one was 69,XXX and the healthy one was 46,XX. CONCLUSION: Early developed growth discordance in a twin pregnancy may be a sign of chromosomal abnormality such as triploidy. Those cases should be karyotyped without delay to decrease preterm delivery risk of possible selective feticide. PMID- 16354979 TI - Prenatal diagnosis of trisomy 9. AB - Fetal trisomy 9, especially its nonmosaic form, is a rare chromosomal abnormality and there are only 8 cases reported to have been sonographically detected in the prenatal period. We report a case of nonmosaic fetal trisomy 9, mimicking trisomy 13 on sonographic findings at 32 weeks' gestation. Although the incidence of trisomy 9 is rare, diagnosing trisomy 9 is important because the sonographic features are similar to those of trisomies 13 and 18, and cannot to be identified by routine fluorescencein situ hybridization. Because nonmosaic trisomy 9 is universally lethal, correct diagnosis and appropriate counseling is essential in patient care and clinical management. PMID- 16354980 TI - Amiodarone in treatment of fetal supraventricular tachycardia. A case report and review of literature. AB - We report a case of nonimmune hydrops fetalis detected at 32 weeks of gestation. Fetal heart rate was 300 beats per minute. Ultrasound and fetal Doppler echocardiography showed it to be due to supraventricular tachycardia (SVT). Following failed maternal therapy with digoxin alone, amiodarone with digoxin was used. Conversion to sinus rhythm and resolution of hydrops followed this treatment. Since there is no ideal treatment protocol for these cases at present, we reviewed reports of transplacental treatment of SVT. PMID- 16354981 TI - Reduced nitric oxide in amniotic fluid of patients with chorioamnionitis. AB - OBJECTIVE: Levels of nitric oxide (NO) and cytokines were assessed in amniotic fluid obtained from patients with severe chorioamnionitis (CAM) and appropriate controls. METHODS: Amniotic fluid was obtained from 12 patients with CAM (17-24 weeks of gestation) and 89 patients undergoing diagnostic amniocentesis (16-18 weeks of gestation). The concentrations of NO, interleukin-6 (IL-6), and leukocyte elastase (LE) in amniotic fluid were then measured and compared. RESULTS: The concentrations of NO, IL-6, and LE were all higher in CAM cases than in normal pregnant women. Furthermore, an inverse correlation between NO and LE was suggested in the CAM group. CONCLUSIONS: These results indicate that in severe CAM, the action of NO might be reduced, not only due to blockage of action but also by degradation, despite increased production. PMID- 16354982 TI - Heterotopic trigeminal pregnancy in infertile women after ovulation stimulation and embolisation of a uterine myoma. AB - Heterotopic pregnancy is a simultaneous occurrence of intra-uterine pregnancy and ectopic pregnancy. The incidence of ectopic pregnancy has increased as a consequence of assisted reproduction and ovulation stimulation agents. In this report, we describe the case of a 34-year-old nulliparous woman who became pregnant after ovulation induction with clomiphene. According to her gynaecologic history, she had embolisation of a uterine myoma. The report presents a case of ectopic and twin intra-uterine pregnancy. After total laparoscopic salpingectomy, she had normal intra-uterine pregnancy. PMID- 16354983 TI - Increased expression of vascular endothelial growth factor in cardiac structures of fetus with hydrops as compared to nonhydropic controls. AB - OBJECTIVE: The hypothesis that severe fetal hydrops is caused by an excess of vascular endothelial growth factor (VEGF), mainly produced in the fetal heart, is tested. METHODS: Immunohistochemical VEGF-stained postmortem biopsies from the right ventricle and right atrium of 8 hydropic fetuses were compared to those of 8 nonhydropic fetuses. The endocardium, myocardium, epicardium, endothelium, and vascular smooth muscle cells were scored on intensity of VEGF-staining. The Mann Witney test was used to test for significancy (p < 0.05) of the differences in staining. Increased vascularization as a result of VEGF was measured in both groups by standard randomization count. RESULTS: The endocardium, epicardium and endothelium of the coronary vessels showed significantly (p < 0.05) more intense VEGF-staining in the hydrops group than in the control group. The atria showed more intense staining than the ventricles in both groups. The hydropic fetuses showed a significantly increased number of coronary vessels in the myocardium. These vessels contained more blood cells than the coronary vessels in nonhydropic fetuses. CONCLUSION: The fetal heart appears to be a major source of excess VEGF in fetal hydrops. PMID- 16354984 TI - Ballantyne syndrome: a case report. AB - Ballantyne syndrome (also called mirror syndrome or triple edema) describes the unusual association of fetal and placental hydrops with maternal preeclampsia. This is a case report illustrating a 37-year-old patient who was referred to our clinics at 28 weeks of gestation (wg) because of fetal hydrothorax. On examination, the woman did not show signs of preeclampsia. The fetal ultrasound examination revealed bulky hydrothorax, generalized subcutaneous edema, placental edema, and polyhydramnios. It was not possible to find the cause of the fetal hydrops. At 29 weeks and 4 days of gestation, the fetal hydrothorax was removed by two pleuro-amniotic shunts, but at the moment of our intervention anasarca was already present. In the following 3 days, despite observing bed rest, the mother developed edema of hands and face, while blood pressure remained normal. At 30 wg the patient underwent cesarean section because fetal movements ceased and the fetal heart rate monitoring showed loss of variability and decelerations. Before dying, the neonate lived for 20 days in a state of deep hypotension. PMID- 16354985 TI - Is the nuchal index increased in fetuses with congenital structural heart defects? AB - OBJECTIVES: To determine if the Nuchal index (NIx) is increased in euploid fetuses with structural congenital heart defects (CHD). METHODS: Euploid fetuses with CHD between 18 and 24 weeks gestation were identified. The next fetus meeting the same criteria with a normal fetal echocardiogram were selected as a control. The NIx [(mean nuchal thickness /mean biparietal diameter) x 100] and cardiac axis (CA; degrees) were calculated for each fetus. Standard descriptive tests and two-tailed t test were used. RESULTS: The NIx in the abnormal (n = 20) and control (n = 20) groups were 9.10 (2.35) and 7.54 (p = 0.04) and CA was 55.8 degrees and 48.6 degrees (p = 0.02), respectively. CONCLUSIONS: The NIx and CA were significantly different in fetuses with CHD. A prospective study to confirm these findings and determine clinical utility is warranted. PMID- 16354986 TI - Pulmonary balloon valvuloplasty in a fetus with critical pulmonary stenosis/atresia with intact ventricular septum and heart failure. AB - Outcome of fetuses with critical pulmonary stenosis (critical PS) or atresia of the pulmonary valve (PA) with intact ventricular septum (IVS) is closely related with right ventricle hypoplasia and its consequent hemodynamics. Fetal echocardiography not only allows early detection of this condition but also monitors its normally unfavorable evolution. These cases may benefit from intrauterine intervention relieving outflow tract obstruction in order to achieve a biventricular circulation. Successful valvuloplasty of the pulmonary valve was performed in a fetus with critical PS-IVS and heart failure at 25 weeks. After the procedure there was a significant improvement in fetal hemodynamics. Follow up scans at 34 weeks detected a significant restenosis with signs of circulatory failure leading to premature delivery of the baby. An immediate postnatal valvuloplasty successfully completed the ultimate objective of biventricular repair. Fetal pulmonary valvuloplasty is feasible and may change the natural history of the disease in fetuses with critical PS-IVS. PMID- 16354987 TI - Three-dimensional cephalometric evaluation of maxillary growth following in utero repair of cleft lip and alveolar-like defects in the mid-gestational sheep model. AB - OBJECTIVE: To evaluate maxillary growth following in utero repair of surgically created cleft lip and alveolar (CLA)-like defects by means of three-dimensional (3D) computer tomographic (CT) cephalometric analysis in the mid-gestational sheep model. METHODS: In 12 sheep fetuses a unilateral CLA-like defect was created in utero (untreated control group: 4 fetuses). Four different bone grafts were used for the alveolar defect closure. After euthanasia, CT scans of the skulls of the fetuses, 3D reconstructions, and a 3D-CT cephalometric analysis were performed. RESULTS: The comparisons between the operated and nonoperated skull sides as well as of the maxillary asymmetry among the experimental groups revealed no statistically significant differences of the 12 variables used. CONCLUSIONS: None of the surgical approaches used for the in utero correction of CLA-like defects seem to affect significantly postsurgical maxillary growth; however, when bone graft healing takes place, a tendency for almost normal maxillary growth can be observed. PMID- 16354988 TI - Abruptio placentae during fetal myelomeningocele repair. AB - Myelomeningocele (MM) is a congenital neural tube defect with serious consequences, including hydrocephaly. An important hope for intrauterine repair is that hydrocephaly may be prevented by reversing the Arnold-Chiari malformation. Three medical centers in the United States are doing trials with this objective. We describe an intrauterine correction of MM in a Brazilian research center of fetal medicine, which resulted in abruptio placentae and fetal death, to illustrate factors that influence fetal-maternal risks during this surgical procedure. PMID- 16354989 TI - Prenatal exclusion/confirmation of Fanconi anemia via flow cytometry: a pilot study. AB - OBJECTIVE: To explore the potential of flow cytometry in the prenatal exclusion or confirmation of Fanconi anemia (FA). METHODS: Indications for prenatal diagnosis were (1) FA-negative family history, but suspicious ultrasound findings such as radial ray aplasia, (2) FA-positive family history, but without knowledge of the affected gene and/or mutation. Amniotic fluid (AF) cell cultures and umbilical cord (UC) blood cultures were assayed for typical cell cycle changes (G2-phase accumulations) without and with mitomycin C (MMC) treatments using single- and dual-parameter (BrdU-Hoechst) flow cytometry. RESULTS: Single parameter flow cytometry correctly identified 2 positive and 9 negative cases on the basis of MMC sensitivity of cultivated AF cells. Likewise, 8 negative and 2 positive cases were correctly predicted using bivariate flow cytometry of 72-hour UC blood cultures. In contrast, bivariate flow cytometry applied to AF cells grown in the presence of bromodeoxyuridine (BrdU) yielded false-positive and false-negative results. CONCLUSIONS: Single-parameter flow cytometry of AF cell cultures and bivariate flow cytometry of UC cell cultures have the potential to correctly predict the affected status in cases at risk for FA, whereas bivariate flow cytometry proved unreliable when applied to BrdU-substituted AF cell cultures. Cases with a low a priori risk (e.g. sonographic finding of radial ray abnormalities and negative family history) would benefit most from flow cytometry as a rapid and economical prenatal screening procedure. PMID- 16354990 TI - Prenatal diagnosis of trisomy 21 with fetal cells in maternal blood using comparative genomic hybridization. AB - OBJECTIVE: This study was undertaken to determine the clinical use of comparative genomic hybridization (CGH) for detection of fetal trisomy 21 from fetal ceIls (nucleated red blood cells; nRBCs) isolated from maternal peripheral venous blood. METHODS: Maternal peripheral venous blood samples were collected in sterile tubes containing heparin. After triple density gradient centrifugation, magnetic activated cell sorting using CD45 and CD71 was used to isolate the fetal nRBCs. Fetal nRBCs were successfully isolated from maternal peripheral blood in all cases. After laser-microdissecting fetal nRBCs, degenerate oligonucleotide primed polymerase chain reaction, and nick translation, DNA size was suitable for hybridization. RESULTS: By CGH analysis, we diagnosed one normal male, one normal female, and one trisomy 21 male fetus. These results were confirmed by amniocentesis. CONCLUSIONS: Prenatal diagnosis from fetal cells in maternal peripheral blood by CGH shows clinical promise as an alternative or as a supplement to fluorescence in situ hybridization with chromosome-specific probes but further studies are warranted. PMID- 16354991 TI - Cardiac circumference measurement: possible screening tool in early pregnancy for anomalous cardiac development. AB - OBJECTIVE: In this study, cardiac circumference (CC), measured by transvaginal sonography, was analyzed to determine the growth during the early stage of pregnancy and its relationship to abdominal circumference and femur length (FL). METHODS: Biometric measurements, including CC, were obtained in 1,182 euploid fetuses at 9-16 weeks' gestation. The CC measurements were related to gestational age (GA), abdominal circumference, and FL. The corresponding 95% confidence intervals were calculated. RESULTS: A linear growth function was observed between CC (mm) and GA (days) (r2 = 0.601; p < 0.0001; y = 0.573 GA - 24.185). Similarly, a good correlation is described with a linear function between CC (mm) and abdominal circumference (mm) (r2 = 0.70; p < 0.0001; y = 0.343 AC + 3.696) and between CC (mm) and FL (mm) (r2 = 0.626; p < 0.0001; y = 1.335 FL + 14.444). The regression analysis that best correlates the dependent variable CC (mm) with the independent variables, GA (days), abdominal circumference (mm), and FL (mm), is: y = 0.137 GA + 0.235 AC + 0.199 FL - 3.303 (r2 = 0.708; p < 0.0001). CONCLUSION: Our results provide normative data of the growth of the CC in early pregnancy. The good correlation described between CC and abdominal circumference and FL suggests that cardiac measurements in early pregnancy alone, or related to other fetal biometric parameters, could be used as a screening tool to identify fetuses at risk for abnormal heart development. PMID- 16354992 TI - Higher median levels of free beta-hCG and PAPP-A in the first trimester of pregnancy in a Chinese ethnic group. Implication for first trimester combined screening for Down's syndrome in the Chinese population. AB - OBJECTIVE: To study the effect of ethnic Chinese on the medians of free beta-hCG and PAPP-A in the first trimester of pregnancy. METHODS: The data of 943 women undergoing first trimester combined screening for fetal Down syndrome were analysed to derive the Chinese-specific medians. The calculated risk of Down syndrome based on these Chinese-specific medians was compared with that based on the original algorithm of the Fetal Medicine Foundation (FMF). RESULTS: The maternal serum levels of multiples of median of free beta-hCG and PAPP-A were significantly higher among the Chinese than among the Caucasians. The weight adjusted gestation-specific medians were developed. Without adjustment for ethnicity, the original FMF algorithm underestimated the risk of Down syndrome by a median of 1%. Adjustment by ethnicity increased the false-positive rate by 10% (from 5.3 to 5.9%). CONCLUSION: Ethnic Chinese have a significantly higher maternal serum level of free beta-hCG and PAPP-A in the first trimester, which could not be explained by differences in maternal weight. Adjustment for ethnicity may be necessary for these biochemical markers in a first trimester screening program. PMID- 16354993 TI - Gender-related differences in fetal heart rate during first trimester. AB - OBJECTIVE: Many expecting parents wish to ascertain fetal gender early in pregnancy. Our goal was to determine whether fetal heart rate (FHR) of males and females during the first trimester is significantly different. MATERIALS AND METHODS: From November 1997 to February 2003 we enrolled pregnant women with singleton gestations who underwent obstetric sonography at less than 14 weeks of gestational age. Indications for the sonographic study included first-trimester bleeding, uncertain gestational dating, poor obstetrical history, and aneuploidy screening by nuchal translucency. The sonographic studies were performed by a single sonographer and reviewed by the first author. The FHR was determined by m mode. All subjects underwent second-trimester sonography at 18.0-24.0 weeks' gestation by the same team, and fetal gender was recorded. Multiple gestations, miscarriages and pregnancies with uncertain fetal gender were excluded. Sonographically assigned fetal gender was confirmed at delivery. RESULTS: Of the 966 first-trimester studies performed, 477 met the inclusion criteria. Of these, 244 (51%) were female and 233 (49%) were males. There were no statistical differences in mean maternal age, gravidity, parity, and mean gestational age at the time of the first study (9.0 +/- 2.3 weeks for female fetuses and 9.0 +/- 2.3 weeks for males, p = 0.7). The average female FHR was 151.7 +/- 22.7 bpm and male FHR was154.9 +/- 22.8 bpm (p = 0.13). DISCUSSION: Contrary to beliefs commonly held by many pregnant women and their families, there are no significant differences between male and female FHR during the first trimester. PMID- 16354994 TI - A folate-dependent metabolite in amniotic fluid from pregnancies with normal or trisomy 21 chromosomes. AB - INTRODUCTION: Previous studies have given conflicting results as to whether or not folate metabolism is altered in Down syndrome. Folate is necessary to facilitate metabolism of one-carbon units. Folate accepts one-carbon units from one-carbon unit donors, including formiminoglutamate (FIGLU). Folate deficiency leads to accumulation of FIGLU and impairment of one-carbon unit metabolism. FIGLU is a functional measure of folate deficiency. MATERIALS AND METHODS: Archived anonymized amniotic fluid specimens were obtained from normal pregnancies and those with Down syndrome. Gas liquid chromatography/mass spectrometry was used to quantitate FIGLU, which is elevated in folate deficiency. A tetra-deuterated FIGLU was used as a standard, and single-ion monitoring was performed. Nonparametric statistical analysis was performed with the Mann-Whitney U test. RESULTS: FIGLU was significantly lower in pregnancies with Down syndrome. The median FIGLU level was 0.9 micromol/l in amniotic fluid from fetuses with Down syndrome. The median FIGLU level was 1.3 in amniotic fluid from control fetuses. This difference was statistically significant (p = 0.009). No statistically significant differences were found with histidine or glutamate. DISCUSSION: There was no evidence of folate deficiency. FIGLU was decreased, not increased. Decreased FIGLU might result from accelerated activity of one or more genes on chromosome 21, by a gene dosage effect. Genes which might explain the reduced FIGLU include one which degrades FIGLU (glutamate formiminotransferase cyclodeaminase), one which participates in purine synthesis, and one which degrades homocysteine (cystathionine-beta-synthase). PMID- 16354995 TI - Elevated interferon-alpha in fetal blood in the prenatal diagnosis of Aicardi Goutieres syndrome. AB - A case of Aicardi-Goutieres syndrome is described in a family with index cases. The diagnosis was made prenatally based on high fetal blood concentration of interferon alpha. The biological measurement could be of interest for further diagnosis of other cases. PMID- 16354996 TI - Examination of maternal plasma erythropoietin and activin A concentrations with regard to circulatory erythroblast levels in normal and preeclamptic pregnancies. AB - OBJECTIVES: Preeclampsia has been shown to be associated with an increased number of fetal and maternal erythroblasts in the maternal circulation, suggesting that preeclampsia involves increased leakage of fetal cells across the placental barrier, as well as increased erythropoiesis. We examined the relationship between circulatory erythroblast levels with maternal plasma concentrations of erythropoietin and activin A. METHODS: In a case-control study, we examined 15 pregnancies affected by preeclampsia and 10 matched controls. Erythroblasts were enriched from maternal blood samples by magnetic cell sorting, enumerated and correlated with corresponding plasma activin A and erythropoietin concentrations. RESULTS: The proportion of erythroblast was elevated in preeclampsia (0.8 vs. 0.1%, p = 0.023). Erythropoietin and activin A concentrations were significantly elevated in preeclampsia (100.4 vs. 44.5 pg/ml, p = 0.023, and 7.4 vs. 1.85 ng/ml, p = 0.029, respectively). Circulatory erythroblast numbers were found to correlate with plasma activin A concentrations (r = 0.76, p = 0.01) in cases with preeclampsia. No such relationship existed for erythropoietin. CONCLUSIONS: Our data suggest that increased concentrations of activin A promote enhanced levels of erythropoiesis in preeclampsia. As the placenta is one of the major sources of activin A in pregnancy, this increase in activin A-dependent erythropoiesis in preeclampsia may be a reflection of an underlying placental hypoxic condition. PMID- 16354997 TI - Application of pharmacokinetics and pharmacodynamics to antimicrobial therapy of community-acquired respiratory tract infections. AB - To achieve bacteriologic and clinical success, sufficient concentrations of antimicrobial at the site of infection must be maintained for an adequate period of time. These dynamics are determined by combining drug pharmacokinetic and pharmacodynamic (PK/PD) data with minimum inhibitory concentrations. Bacteriologically confirmed failures have been reported in otitis media and, with a lesser degree of evidence, in pneumococcal pneumonia with a variety of agents that include beta-lactams, macrolides and fluoroquinolones. These failures have been shown to be due to infection by resistant pathogens or suboptimal therapy. However, no clinical failure has been reported during therapy for bacteremic pneumococcal pneumonia with adequate doses of beta-lactams. The failures reported with macrolides or fluoroquinolones have been due to either preexisting resistance to these agents that cannot be overcome by increasing the dose of the antimicrobial or, more rarely, the emergence of resistance during therapy. In this review, we offer an overview of the most important attributes of the main antimicrobials that are currently used in the treatment of community-acquired respiratory tract infections from a PK/PD perspective. PMID- 16354998 TI - More than measures and mechanics. The art of breathing. PMID- 16354999 TI - Chronic obstructive pulmonary disease beyond cigarette smoke. PMID- 16355000 TI - Vascular endothelial growth factor in the human diaphragm: new insight into adaptation mechanisms in chronic obstructive pulmonary disease patients. PMID- 16355001 TI - Relationship of allergic degree and PC20 level in adults with positive methacholine challenge test. AB - BACKGROUND: The relationship between allergic markers and PC(20) (provocative concentration causing a 20% fall in FEV(1)) in adults with positive methacholine challenge test (MCT) is still subject to discussion. OBJECTIVE: The goal of this study is to determine whether the degree of bronchial hyperresponsiveness (BHR) is correlated with the severity of atopy or not. METHODS: Forty patients with persistent cough and/or unexplained episodes of dyspnea exhibiting a normal baseline pulmonary function test and positive MCT were enrolled in this study. They were divided into two groups according to their Phadiatop response (positive and negative), and three groups by PC(20) level: PC(20) < 1.25 mg/ml, 1.25 mg/ml < or = PC(20) < 5 mg/ml, and 5 mg/ml < or = PC(20) < 25 mg/ml. RESULTS: Twenty three patients (57.5%) were Phadiatop positive. Of the 23 Phadiatop-positive patients, 20 patients (87%) were positive for specific serum IgE to Dermatophagoides pteronyssinus (D1) and Dermatophagoides farinae (D2). The total serum IgE and eosinophil cationic protein (ECP) level of the Phadiatop-positive group were higher than those of the Phadiatop-negative group (p < 0.0001 and p = 0.005, respectively). The age, gender, FEV(1)/FVC ratio, ECP and total serum IgE level were not significantly different in the three PC(20) subgroups. The Phadiatop response and PC(20) level were significantly correlated (p = 0.0063). For Phadiatop-positive adults, PC(20) level was not significantly correlated with total serum IgE and specific serum IgE to D1 and D2. CONCLUSION: Atopy is related to adults with positive MCT. However, the degree of BHR to methacholine is not significantly correlated with the severity of atopy. PMID- 16355002 TI - Sedative drug requirements during flexible bronchoscopy. AB - BACKGROUND: There is a paucity of data comparing doses of sedative medication during bronchoscopy in immunosuppressed and non-immunosuppressed patients. OBJECTIVES: The aim of this study was to define the sedative medication doses used in specific patient groups during bronchoscopy. METHODS: Bronchoscopy was performed under local anesthesia, sedation with intermittent boluses of intravenous midazolam and intravenous hydrocodone 5 mg. Two hundred and thirty nine consecutive bronchoalveolar lavage procedures were included. Procedures in non-immunosuppressed patients were classified as controls (n = 91). Procedures in immunosuppressed patients who received midazolam consisted of stem cell transplant (34), solid organ transplant (25), chemotherapy (33), HIV with drug abuse (10), HIV (5), prednisone (17) and immunosuppression for other diseases (12). Intravenous propofol was administered during 12 procedures due to inability to achieve optimal sedation with midazolam in a previous bronchoscopy (stem cell transplant recipient 1, lung transplant for cystic fibrosis 5) and during the same bronchoscopy due to inadequate sedation with a high dose of midazolam--renal transplant recipient 1, drug abuse (HIV 1, renal transplant recipient 1), bronchoscopy combined with gastroscopy (2) and a hypoxemic patient (1). The mean dose of propofol administered was 2.8 +/- 1.3 mg/kg. RESULTS: Midazolam requirement was significantly higher in patients with stem cell transplantation (0.09 +/- 0.05 mg/kg) compared with controls (0.06 +/- 0.03 mg/kg; p = 0.0002). In the HIV patients with drug abuse (0.12 +/- 0.10 mg/kg), there was a tendency for the need of a higher dose of midazolam compared with the control group (p = 0.0754). CONCLUSION: Stem cell transplant recipients and selected HIV patients with drug abuse need higher doses of midazolam for bronchoscopy. PMID- 16355003 TI - Management of benign stenoses of the large airways in the university hospital in Prague, Czech Republic, in 1998-2003. AB - BACKGROUND: Clinically significant benign stenoses of the large airways develop in about 1% of patients after intubation. The management of benign stenoses is not unified around the world, nor are there any accepted methods for their screening. OBJECTIVES: The purpose of this study is to describe and compare results of interventional bronchoscopy and surgical therapy of benign stenoses as well as to propose an algorithm for the management of this airways disorder. METHODS: Prospective study on 80 consecutive patients with benign stenoses of the large airways admitted to the Pulmonary Department of the University Hospital of Prague-Motol. RESULTS: Sixty-two patients developed stenoses after endotracheal intubation or tracheostomy, in 18 patients the stenosis was caused by other diseases or pathological situations. Thirty-eight patients were sent for surgical resection of the stenotic part of the airways. 2 surgically treated patients developed recurrence of the stenosis and had to be reoperated on. Narrowing of the trachea at the site of end-to-end anastomosis developed in 6 other patients and was cured by interventional bronchoscopy. The remaining 42 patients were treated by interventional bronchoscopy (Nd-YAG laser, electrocautery, stent) which was curative in 35 patients. Sixty-five patients were alive at the time of evaluation, 15 patients died. Five of them died between 3 and 14 (median 4) months after surgery from a disease other than airway stenosis. Ten nonresected patients also died, with 1 exception, due to a disease other than airway stenosis; the median survival was 9 months. CONCLUSIONS: We recommend to assess the patient for surgery after the initial diagnosis and therapeutic bronchoscopy with dilatation of the stenosis. If the patient is not a suitable candidate for resection, interventional bronchoscopy is an appropriate alternative for the management of benign stenoses of the large airways. PMID- 16355004 TI - Adenoviral E1A suppresses secretory leukoprotease inhibitor and elafin secretion in human alveolar epithelial cells and bronchial epithelial cells. AB - BACKGROUND: An imbalance between neutrophil protease and surrounding antiprotease levels has been shown to be important in the pathogenesis of chronic obstructive pulmonary disease (COPD). Adenoviral E1A DNA and protein are frequently detected in the lungs of COPD patients. As secretory leukoprotease inhibitor (SLPI) and elafin/skin-derived antileukoproteinase (SKALP) are locally produced in the lung and inhibit neutrophil elastase activity, we hypothesized that adenoviral E1A might affect the production of these antiproteases. OBJECTIVES: To examine the effect of E1A on SLPI and elafin/SKALP secretion in A549 (alveolar epithelial) cells and primary human bronchial epithelial (HBE) cells. METHODS: SLPI and elafin/SKALP were quantitated from cell culture supernatants using an ELISA. SLPI mRNA expression was examined by Northern blotting, and SLPI promoter activity was measured using a reporter gene assay. RESULTS: E1A significantly suppressed SLPI and elafin/SKALP secretion by A549 cells upon interleukin (IL)-1beta stimulation. E1A also suppressed SLPI and elafin/SKALP secretion by HBE cells. SLPI mRNA expression in A549 cells was suppressed by E1A regardless of IL-1beta stimulation. IL-1beta-induced SLPI promoter activity was suppressed by E1A gene transfection into A549 cells. CONCLUSIONS: Our findings of adenoviral E1A mediated suppression of SLPI and elafin/SKALP secretion suggest that E1A may be involved in the enhancement of alveolar damage and play a role in the COPD process. PMID- 16355005 TI - Vascular-specific growth factor mRNA levels in the human diaphragm. AB - BACKGROUND: Angiogenesis is an adaptation mechanism of skeletal muscles to increased load. Animal data have shown increased vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and transforming growth factor-beta(1) (TGF-beta(1)) mRNA levels in the diaphragm as a result of increased minute ventilation, but there are no data concerning the human diaphragm. OBJECTIVES: The purpose of this study was to investigate the VEGF, bFGF, TGF-beta(1) mRNA levels in the human diaphragm of normal subjects and patients with altered respiratory mechanics. METHODS: We studied 9 patients with chronic obstructive pulmonary disease (COPD), 4 obese patients and 12 controls. We performed multiplex semiquantitative reverse transcription polymerase chain reaction to determine the VEGF, bFGF and TGF-beta(1) mRNA levels in specimens taken from their diaphragm. RESULTS: VEGF mRNA levels were 18% higher in COPD patients compared with controls (p = 0.04), while for the obese patients, these levels were not statistically significantly different. bFGF and TGF-beta(1) mRNA levels in COPD patients or obese individuals compared with controls did not differ significantly either. CONCLUSIONS: The results of our study showed that TGF-beta(1), VEGF and bFGF mRNA was detected in the human diaphragm. The VEGF levels were higher in COPD patients than in normal subjects. This upregulation of VEGF may suggest an enhancement of angiogenesis in the diaphragm in COPD patients. PMID- 16355006 TI - Two full-term pregnancies in a patient with mitochondrial myopathy and chronic ventilatory insufficiency. AB - Mitochondrial myopathies are a group of diseases characterized by metabolic defects at the mitochondrial respiratory chain level which result in impaired oxidative phosphorylation and ATP synthesis. As with other neuromuscular diseases, respiratory muscles can be affected and ventilatory failure may occur. There have been isolated case reports of pregnant patients with ventilatory failure due to neuromuscular diseases such as polio and spinal muscular atrophy. We describe the case of a 34-year-old patient with mitochondrial myopathy and ventilatory failure requiring non-invasive ventilation who carried two pregnancies to term with no complications. We have not found a similar case in the literature. PMID- 16355007 TI - Leukopenia during treatment with first-line anti-tuberculosis medication. PMID- 16355008 TI - The metabolic syndrome: an overview. PMID- 16355009 TI - Metabolic syndrome: historical perspectives. AB - The metabolic syndrome is an aggregation of biochemical and physical conditions that presage the development of atherosclerotic cardiovascular disease. The history of the metabolic syndrome is rooted in the recognition of adipose tissue as a heterogeneous, biologically active organ, as well as in the concepts of insulin resistance and its consequences. Establishment of the metabolic syndrome as a disease entity has been hindered by non-uniform criteria for its diagnosis. PMID- 16355010 TI - Definition of the metabolic syndrome: current proposals and controversies. AB - Metabolic syndrome includes a clustering of metabolic derangements that cause affected subjects to have an increased risk for developing diabetes, cardiovascular disease, and, according to recent epidemiologic studies, chronic kidney disease. The present review discusses four definitions of metabolic syndrome published by different national and international committees. In an effort to bridge the differences existent in those classifications, a unified definition that recognizes the increased biologic activity of the upper visceral fatty tissue and the strong association of abdominal obesity as a leading part of metabolic syndrome is proposed herein. The diagnosis of metabolic syndrome is reserved for pre-diabetic patients who share the risk of becoming diabetic or developing cardiovascular or chronic kidney disease. PMID- 16355011 TI - Epidemiology of the metabolic syndrome. AB - The clustering of cardiovascular risk factors, known as the metabolic syndrome, greatly increases the risk of developing diabetes, kidney disease, and cardiovascular disease. Individuals with the metabolic syndrome are also at increased risk for premature death from cardiovascular disease or all-cause mortality. Cross-sectional and longitudinal epidemiologic studies provide prevalence data on the syndrome based on criteria proposed by the World Health Organization and the National Cholesterol Education Program Adult Treatment Panel III. Owing to differences in the criteria, estimates of the prevalence of the syndrome vary according to the criteria used. Generally, the syndrome is more common in older people and in the United States and it is more prevalent among Mexican Americans. Obesity and sedentary lifestyles are major contributing factors to the syndrome and provide opportunities for interventions. Recent data from a randomized controlled trial indicate that a weight loss and exercise intervention reduced the incidence of the metabolic syndrome by 41% among individuals with impaired glucose tolerance. Pharmacologic treatment of the individual components of the metabolic syndrome provides an alternate strategy for managing the syndrome. The rising global epidemics of overweight and obesity will likely lead to increases in the prevalence of the metabolic syndrome posing a serious burden for clinicians and public health officials. PMID- 16355012 TI - Fat as an endocrine organ: relationship to the metabolic syndrome. AB - Obesity and the metabolic syndrome have both reached pandemic proportions. Together they have the potential to impact on the incidence and severity of cardiovascular pathologies, with grave implications for worldwide health care systems. The metabolic syndrome is characterized by visceral obesity, insulin resistance, hypertension, chronic inflammation, and thrombotic disorders contributing to endothelial dysfunction and, subsequently, to accelerated atherosclerosis. Obesity is a key component in development of the metabolic syndrome and it is becoming increasingly clear that a central factor in this is the production by adipose cells of bioactive substances that directly influence insulin sensitivity and vascular injury. In this paper, we review advances in the understanding of biologically active molecules collectively referred to as "adipokines" and how dysregulated production of these factors in obese states mediates the pathogenesis of obesity associated metabolic syndrome. PMID- 16355013 TI - The key role of insulin resistance in the cardiometabolic syndrome. AB - Insulin resistance is invariably present in patients with the cardiometabolic syndrome and is thought to play a key role in its pathogenesis. It represents a complex interaction of maladaptive characteristics related to impaired insulin action at target organs and external factors such as genetics and environment. It is likely that the molecular factors that underlie insulin resistance and resultant hyperinsulinemia contribute to many of the clinical components of the cardiometabolic syndrome, although the precise associations remain poorly understood. Abnormalities of various adipocytokines (particularly reduced circulating levels of adiponectin and leptin) resistance may also play a role in the genesis of insulin resistance. Genetic factors are an important determinant of insulin sensitivity. Effective management of the cardiometabolic syndrome will ultimately depend on our understanding of the underlying molecular mechanisms of key components of this disorder, such as insulin resistance. PMID- 16355014 TI - Importance and management of dyslipidemia in the metabolic syndrome. AB - The metabolic syndrome is a common disorder characterized by central obesity, impaired glucose tolerance, hypertension, and atherogenic dyslipidemia (including the combination of hypertriglyceridemia, low levels of high-density lipoprotein cholesterol, and a preponderance of small, dense low-density lipoprotein particles). In this manuscript, we review the pathogenesis and significance of dyslipidemia in the metabolic syndrome, the role of nonpharmacologic therapy with therapeutic lifestyle changes, and drug therapies, including statins, fibrates, nicotinic acid, and omega-3 fatty acids or fish oils, alone or in drug combinations, to improve lipids and reduce the chance of subsequent cardiovascular disease events. PMID- 16355015 TI - Hypertension and the metabolic syndrome. AB - The cause of hypertension in the metabolic syndrome is complex and multifactorial and all of the elements of the metabolic syndrome, including obesity, insulin resistance, and the characteristic dyslipidemia probably are involved in mediating changes ultimately resulting in hypertension and modifying its course. Of these elements, obesity may play the most important and pivotal role in creating the conditions that lead to hypertension in the metabolic syndrome. This is not to say that the other elements of the syndrome are less important, and, as we gain more insight into the processes involved, we should be able to better manage the disease and tailor our therapeutic interventions appropriately. PMID- 16355016 TI - The cardiometabolic syndrome as a cardiovascular risk factor. AB - The cardiometabolic syndrome (CMS) is associated with cardiovascular disease (CVD) and includes a constellation of risk factors such as central obesity, hypertension, insulin resistance, dyslipidemia, microalbuminuria, and hypercoagulability. Collectively, these risk factors increase CVD endpoints such as stroke, congestive heart failure, chronic kidney disease (CKD), and overall mortality. The CMS is associated with endothelial dysfunction, inflammation, abnormal thrombolysis, and increased oxidative stress that accentuate progression of CVD. We will review how the varying components of the CMS relate to an increased CVD and renal disease risk. PMID- 16355017 TI - Kidney disease and the metabolic syndrome. AB - The epidemic of metabolic syndrome contributes to the rapid growth of cardiovascular and renal diseases. Hyper-hemodynamics, impaired pressure natriuresis, excess excretory load, insulin resistance, endothelial dysfunction, chronic inflammation, and prothrombotic status individually and interdependently initiate renal injury in metabolic syndrome. The prevention and treatment of kidney disease require a multifactorial approach. Weight loss through diet control and exercise can reverse many pathophysiologic processes. Pharmacologic intervention includes insulin sensitizers, tight glycemic and lipid control, blockage of renin angiotensin aldosterone system, and anti-inflammatory and antithrombotic therapies. Each peroxisome proliferator-activated receptor isoform plays a distinct role in metabolic syndrome, and their agonists may prevent or reverse the early renal injuries. PMID- 16355018 TI - Nonalcoholic steatohepatitis and the metabolic syndrome. AB - Relatively recently, the liver has been recognized as a major target of injury in patients with insulin resistance or the metabolic syndrome. Insulin resistance is associated with fat accumulation in the liver, a condition called nonalcoholic fatty liver disease (NAFLD). Excess fat in the liver is not a benign condition. Some patients with NAFLD develop necroinflammatory changes in the liver called nonalcoholic steatohepatitis (NASH) and a fraction of those will develop cirrhosis. About 20% all adults have NAFLD and 2% to 3% of adults have NASH. Approximately 20% of patients with NASH are at risk for developing cirrhosis and subsequently dying from end-stage liver disease. The diagnosis of NASH requires a high index of suspicion, especially in obese patients over the age of 45 years who have diabetes, because these are the patients at greatest risk for developing cirrhosis. Treatment focuses on addressing the underlying insulin resistance with increased exercise and weight reduction. PMID- 16355019 TI - Polycystic ovarian syndrome and the metabolic syndrome. AB - Polycystic ovarian syndrome (PCOS), first described in 1937, was defined by specific ovarian histopathology and a constellation of signs and symptoms. Through the years, the etiology remained elusive, with heated debates focusing in turn on the ovary and then the pituitary as the causative agents. In the last several decades, it has become clear that insulin resistance makes up a very important component of this syndrome. With this knowledge, new therapies have emerged along with the realization that PCOS and the metabolic syndrome are closely related through their shared insulin resistance. In this review, the diagnosis, pathophysiology, and therapy of PCOS are discussed and upon this background, those areas held in common by PCOS and the metabolic syndrome are explored. PMID- 16355020 TI - Management of the metabolic syndrome. AB - The metabolic syndrome is most commonly defined as the presence of three or more of the following atherogenic risk factors: increased abdominal waist circumference, hypertriglyceridemia, low serum high-density lipoprotein level, raised blood pressure, and a fasting blood sugar level greater than 110 mg/dL. It is associated with increased risk for the development of atherosclerosis and cardiovascular disease. Current management recommendations promote therapeutic lifestyle changes (mainly diet and exercise) for all of the core elements of the syndrome. Pharmacologic therapy is indicated in most patients with atherogenic dyslipidemia and in some with hyperglycemia and raised blood pressure. These therapeutic measures are able to favorably modify the core components of the metabolic syndrome and it is hoped that they will also improve the long-term cardiovascular prognosis in patients with this disorder. PMID- 16355022 TI - Diagnosis and management of the metabolic syndrome: an American Heart Association/National Heart, Lung, and Blood Institute scientific statement. PMID- 16355023 TI - Selective site ventricular pacing. AB - PURPOSE OF REVIEW: The traditional site for ventricular pacing, the right ventricular apex, produces an abnormal pattern of ventricular depolarization and there is growing evidence that pacing from this site is associated with adverse functional and structural changes in the left ventricle. This is manifest clinically as an increased morbidity and mortality. These observations have fuelled interest in pacing at sites alternative to the right ventricular apex. In this article, we review the evidence for selective site pacing, focusing mainly on the right ventricular outflow tract. RECENT FINDINGS: Data are conflicting on the acute and medium-term effects of right ventricular outflow tract pacing. Although a recent meta-analysis has suggested acute benefit from pacing at this site, the data are confounded by poor definition of the outflow tract and the non randomized nature of most trials. There is a need for standardization of nomenclature and better definition of non-apical sites. Long-term data on chronic pacing are limited, with two studies showing equivalency between apical and outflow tract pacing. In another two studies, right ventricular outflow tract pacing was associated with improved ejection fraction. SUMMARY: Selective site pacing holds promise in attempting to reduce the problems associated with chronic ventricular pacing. Large, prospective, randomized control trials are needed. PMID- 16355024 TI - Recent primary prevention implantable cardioverter defibrillator trials. AB - PURPOSE OF THE REVIEW: The aim of this article is to summarize the most relevant findings of recently published trials on prophylactic implantable cardioverter defibrillator therapy. RECENT FINDINGS: A number of important randomized clinical trials on the efficacy of prophylactic implantable cardioverter defibrillator therapy in patients deemed to be at high risk for ventricular tachyarrhythmias have recently reported their results. Patients with chronic ischemic cardiomyopathy, a long history of heart failure, and an ejection fraction of 0.30 or below benefit from preventive device therapy and are thus candidates for prophylactic defibrillator implantation. For this purpose, a single chamber device appears to be appropriate since there have been no prospective studies showing convincing clinical benefit by adding an atrial lead. Prophylactic implantable cardioverter defibrillator therapy should not be used in patients with recent myocardial infarction. There is convincing evidence from one trial that benefit from the defibrillator in coronary patients accrue after a considerable time has elapsed from the most recent infarct, presumably at least 6 months or perhaps longer. Finally, in patients with chronic dilated non-ischemic cardiomyopathy and a left ventricular ejection fraction of 0.35 or below, there is also benefit from prophylactic implantable cardioverter defibrillator therapy. SUMMARY: Taken together, these trials allow an evidence-based approach to primary prevention of sudden cardiac death in patients with both ischemic and non ischemic cardiomyopathy. PMID- 16355025 TI - The problem of non-response to cardiac resynchronization therapy. AB - PURPOSE OF REVIEW: Cardiac resynchronization therapy improves quality of life, exercise performance, left ventricular ejection fraction, and reduces heart failure hospitalizations and mortality in patients with New York Heart Association class III or IV congestive heart failure and intraventricular conduction delay. A number of key clinical research questions remain, perhaps most importantly the issue of why apparently suitable patients do not respond to cardiac resynchronization therapy. These issues are also relevant to patients who do respond to cardiac resynchronization therapy as potentially their response might be further increased. This article will review the data regarding the frequency of the problem of non-response to cardiac resynchronization therapy and then discuss the postulated reasons and potential solutions. RECENT FINDINGS: Rates of non-response to cardiac resynchronization therapy are often quoted as 20 30%, but a critical analysis of the data would suggest the true non-responder rate can be estimated as perhaps 40-50%. The data indicate that on a population basis non-response is multi-factorial and the extent of mechanical dyssynchrony, left ventricular pacing site and cause of congestive heart failure are likely to be important. Ongoing research is exploring the utility of various techniques for quantifying mechanical dyssynchrony and the potential benefits of targeted left ventricular lead placement and post-implant optimization. SUMMARY: Cardiac resynchronization therapy is a major breakthrough in treatment for advanced congestive heart failure patients. There is substantial rate of non-response to this therapy, however, and research is exploring various ways to increase the response to the technique. PMID- 16355026 TI - Cardiac rhythm management: the shape of things to come. AB - PURPOSE OF REVIEW: The aim of this article is to forecast a major imminent change in the clinical practice of cardiac rhythm management, which is argued to be remote patient monitoring, its potential benefits to clinical practitioners and its barriers to widespread diffusion. RECENT FINDINGS: All four major manufacturers of cardiac rhythm management devices have recently introduced varying types of systems that allow remote patient monitoring. These remote patient monitoring systems promise more efficient patient management in today's clinical setting of rapidly growing numbers of patients with increasingly heterogeneous etiology, varying indications and comorbidities. The major differences between current systems are related to the degree of patient involvement in remote patient monitoring and limits on patient mobility. Other important factors that influence the value of remote patient monitoring are the degree to which remote follow-up would completely fulfill the requirements of an in-office follow-up and the inclusion of sensors that enable the prediction of major clinical events such as heart failure decompensation with a high degree of accuracy. SUMMARY: Even if the different remote patient monitoring systems currently available offer several clinical benefits such as early detection of cardiac events and complications, reduced follow-up costs and increased safety, the full potential of such systems requires the possibility to easily transfer relevant patient data to common patient databases that are linked to hospital information systems or electronic patient records. Only then will it be possible to gain a complete picture of patient conditions. This will require the development of common protocols for data communication and may involve issues of patient data ownership and integrity. PMID- 16355027 TI - Investigating syncope: a review. AB - PURPOSE OF REVIEW: This review focuses on recent literature on the cardiovascular investigation of syncope. RECENT FINDINGS: Syncope is a common and complex clinical entity with many varied etiologies, the diagnosis of which can often be elusive. Recent advances in the area of investigation in syncope include improvements in technologies for arrhythmia event monitoring and an increase in applicability and efficacy of traditional investigations. These advances have increased our ability to manage syncope. SUMMARY: A better understanding of the etiology of syncope in certain cohorts has allowed tailoring of investigations and management of syncope. This is evident when syncope occurs in the presence of structural heart disease, which is associated with a higher incidence of arrhythmias and an increased 1-year mortality. Patients with left-ventricular dysfunction should be considered candidates for an implantable cardioverter defibrillator based on heart function and syncopal presentation. In the absence of significant heart disease, investigations need to be tailored to diagnose neurally mediated causes for syncope and detection of intermittent bradycardia. The improved understanding of syncope has arisen from studies using improved investigational tools for syncope, in particular loop recorders, which are able to monitor cardiac rhythm over long periods. In those without structural heart disease and undifferentiated syncope, neurally mediated causes should be pursed initially. Tilt testing and loop recorders are employed most often in this cohort. Electrophysiological studies have largely been supplanted by implantable cardioverter defibrillator use in structural heart disease, with a low yield in patients with preserved heart function. PMID- 16355028 TI - Genetic profiling as a marker for risk of sudden cardiac death. AB - PURPOSE OF REVIEW: Clinical trials provide evidence that an empirical approach of implantable cardioverter-defibrillator implantation in all heart failure patients (ejection fraction 0.05). CONCLUSION: The radiation dose can be reduced by 50% without loss of nodule detectability by applying the adaptive noise reduction filter to simulated and patient liver images obtained at MDCT. PMID- 16355034 TI - Arterial distribution of calibrated tris-acryl gelatin and polyvinyl alcohol microspheres in a sheep kidney model. AB - OBJECTIVE: The objective of this study was to compare the repartition in the renal arterial vasculature of tris-acryl gelatin microspheres (TGMS) and polyvinyl alcohol microspheres (PVAMS) of 3 calibers (500-700, 700-900, and 900 1200 microm). MATERIALS AND METHODS: Twelve kidneys from 6 adult sheep were embolized and histologically analyzed. The number and size of microspheres and vessels were measured, as well as the deformation of TGMS and PVAMS, and the histologic location according to a classification in 5 zones of the kidney. RESULTS: Two hundred eighty-four vessels were measured. The diameter of the occluded vessels increased when the caliber used for embolization was larger for TGMS and for PVAMS (P < 0.0001, each). The location of TGMS and PVAMS within the vasculature was different for each caliber, because PVAMS blocked significantly more distally than TGMS (P < 0.0001 each). The deformation within the tissue was greater for PVAMS (18.0 +/- 12.3%) than for TGMS (9.0 +/- 8.3%) in general (P < 0.001) and for each caliber of injected microspheres (P < 0.001 each). CONCLUSION: The repartition of a spherical embolic agent in a given vascular network can be influenced by its size and also by its deformation within the vascular bed. PMID- 16355035 TI - Blood flow changes in hepatocellular carcinoma after the administration of thalidomide assessed by reperfusion kinetics during microbubble infusion: preliminary results. AB - OBJECTIVES: We sought to investigate whether thalidomide is able to produce tumor vascular changes in patients with untreatable hepatocellular carcinoma (HCC) that can be detected using microbubble contrast agents. MATERIALS AND METHODS: Eleven consecutive patients with untreatable HCC underwent contrast-enhanced ultrasound before and during thalidomide administration. Real-time destruction reperfusion kinetics was obtained from a representative HCC nodule and from the surrounding liver parenchyma during SonoVue infusion (Bracco, Milan, Italy) at a constant rate of 0.10 mL/s by using a syringe pump and modelized according to the mathematical function SI = A(1 - exp(-betat)) where the plateau signal intensity A reflects the percent blood volume, the time constant beta reflects the average speed of blood, and their product A*beta reflects the nutrient blood flow. RESULTS: Size of the representative nodule reduced significantly 3 to 6 months after the start of thalidomide treatment. Before thalidomide administration A, beta, and A*beta of the index lesion were 44 +/- 60 LIU, 0.31 +/- 0.40 seconds and 8.1 +/- 11.8 LIU/s, respectively). A and A*beta reduced significantly after 15 days (26 +/- 50 LIU and 2.9 +/- 4.8 LIU/s, P < 0.01), 3 months (12 +/- 18 LIU, and 4.3 +/- 7.7 LIU/s, P < 0.01), and 6 months (13 +/- 23 LIU and 2.4 +/- 3.7 LIU/s, P < 0.05) of treatment. No statistically significant changes of the exponential time constant beta were observed, nor changes of A, beta and A*beta in the liver parenchyma. CONCLUSIONS: Contrast-enhanced ultrasound can be used effectively to evaluate changes in perfusion parameters of HCC nodules during thalidomide administration. PMID- 16355036 TI - 64- Versus 16-slice CT angiography for coronary artery stent assessment: in vitro experience. AB - OBJECTIVES: We sought to assess the visualization of different coronary artery stents and the delineation of in-stent stenoses using 64- and 16-slice multidector computed tomography (MDCT). MATERIALS AND METHODS: A total of 15 different coronary stents with a simulated in-stent stenosis were placed in a vascular phantom and scanned with a 16-slice and a 64-slice MDCT at orientations of 0 degree, 45 degrees, and 90 degrees relative to the scanner's z-axis. Visible lumen diameter and attenuation in the stented and the unstented segment of the phantom were measured. Three readers assessed stenosis delineation and visualization of the residual lumen using a 5-point scale. RESULTS: Artificial lumen narrowing (ALN) was significantly reduced with 64-slice CT compared with 16 slice CT. At an angle of 0 degree, 45 degrees, and 90 degrees relative to the scanner's z-axis, the ALN for 16-slice CT was 42.2%, 39.8%, and 44.0% using a slice-thickness of 1.0 mm and 40.9%, 40.4%, and 41.6% using a slice thickness of 0.75 mm, respectively. With 64-slice CT, the ALN was 39.1%, 37.3%, and 36.0% at the respective angles. The differences between attenuation values in the stented and unstented segment of the tube were significantly lower for 64-slice CT. Mean visibility scores were significantly higher for 64-slice CT. CONCLUSION: Use of the 64-slice CT results in superior visualization of the stent lumen and in-stent stenosis compared with 16-slice CT, especially when the stent is orientated parallel to the x-ray beam. PMID- 16355037 TI - Reproducibility of computer-aided volumetry of artificial small pulmonary nodules in ex vivo porcine lungs. AB - OBJECTIVE: The main purpose of this study was to investigate the reproducibility of computed tomography (CT)-based volumetric measurements of small pulmonary nodules. METHODS: We implanted 70 artificial pulmonary nodules in 5 ex vivo porcine lungs in a dedicated chest phantom. The lungs were scanned 5 times consecutively with multislice-CT (collimation 16 x 0.75 mm, slice thickness 1 mm, reconstruction increment 0.7 mm). A commercial software package was used for lesion volumetry. The authors differentiated between intrascan reproducibility, interscan reproducibility, and results from semiautomatic and postprocessed volumetry. RESULTS: Analysis of intrascan reproducibility revealed a mean variation coefficient of 6.2% for semiautomatic volumetry and of 0.7% for human adapted volumetry. For interscan reproducibility a mean variation coefficient of 9.2% and for human adapted volumetry a mean of 3.7% was detected. CONCLUSION: The presented volumetry software showed a high reproducibility that can be expected to detect nodule growth with a high degree of certainty. PMID- 16355038 TI - Computerized quantification of joint space narrowing and periarticular demineralization in patients with rheumatoid arthritis based on digital x-ray radiogrammetry. AB - OBJECTIVES: The aim of our work was to evaluate digital x-ray radiogrammetry (DXR) for the quantification of disease-related periarticular demineralization and computerized analysis of joint space distances (JSDA) for the measurement of joint space narrowing as a new diagnostic method for the early detection of joint associated alterations and for monitoring disease progression in patients with rheumatoid arthritis (RA). MATERIALS AND METHODS: Digital radiographs in 313 patients with varying severity of RA were performed annually and assessed by 2 radiologists using modified Larsen and also the Sharp scores within an observation period of 3 years. The hand radiographs underwent measurements of bone mineral density (BMD) and metacarpal index (MCI) by DXR, as well as computerized JSDA at the metacarpal-phalangeal articulation (JSD-MCP) for a cross sectional and longitudinal study design. RESULTS: Both DXR-BMD (-29.6%; P < 0.01) and DXR-MCI (-31.0%; P < 0.01) revealed a notable reduction dependent on the severity of RA (from grade 1 to grade 5 of the modified Larsen score); the severity dependent decrease of mean JSD-MCP ranged from -31.9% (P < 0.01; Sharp erosion part) to -39.1% (P < 0.01) for the modified Larsen score. Over an observation period of 3 years, a significant decrease of DXR-BMD (-22.3%) and DXR MCI (-23.3%) as well as JSD-MCP mean (-17.5%) was observed (P < 0.05), whereas an accentuated decline of DXR and JSDA parameters was verified for patients without disease-modifying antirheumatic drugs or methotrexate therapy. CONCLUSION: Computerized analysis of hand radiographs by DXR and JSDA is a promising approach to assess the severity and to monitor the progression of RA because DXR and JSDA are timely able to measure periarticular demineralization and also narrowing of JSD-MCP dependent on the severity, the medical treatment and the course of RA. PMID- 16355040 TI - Time continuous tracking and segmentation of cardiovascular magnetic resonance images using multidimensional dynamic programming. AB - In this article, we propose a semiautomatic method for time-continuous contour detection in all phases of the cardiac cycle in magnetic resonance sequences. The method is based on multidimensional dynamic programming. After shape parameterization, cost hypercubes are filled with image-feature derived cost function values. Using multidimensional dynamic programming, an optimal path is sought through the sequence of hypercubes. Constraints can be imposed by setting limits to the parameter changes between subsequent hypercubes. Quantitative evaluation was performed on 20 subjects. Average border positioning error over all slices, all phases and all studies, was 1.77 +/- 0.57 mm for epicardial and 1.86 +/- 0.59 mm for endocardial contours. The average error in end-diastolic and end-systolic volumes over all studies was small: 4.24 +/- 4.62 mL and -4.36 +/- 4.26 mL, respectively. The average error in ejection fraction was 4.82 +/- 3.01%. The reported results compare favorable to the best-reported results in recent literature, underlining the potential of this method for application in daily clinical practice. PMID- 16355039 TI - Ultrasmall supraparamagnetic iron oxide-enhanced magnetic resonance imaging of antigen-induced arthritis: a comparative study between SHU 555 C, ferumoxtran-10, and ferumoxytol. AB - OBJECTIVES: We sought to compare the ability of 3 ultrasmall superparamagnetic iron oxides (USPIOs) to detect and characterize antigen-induced arthritis with MR imaging. MATERIALS AND METHODS: A monoarthritis was induced in the right knee of 18 rats. The left knee served as a normal control. Knees underwent magnetic resonance (MR) imaging before, up to 2 hours, and 24 hours after injection (p.i.) of 200 mumol Fe/kg SHU 555 C (n= 6), ferumoxtran-10 (n = 6), or ferumoxytol (n = 6), using T2-2D-SE 100/20,40,60,80/90 (TR/TE/flipangle), T2*-3D-spoiled gradient recalled (SPGR) 100/15/38, and T1-3D-SPGR 50/1,7/60 sequences. RESULTS: Quantitative signal to noise ratio and DeltaSI data of arthritic knees on T1- and T2*-weighted MR images showed no significant differences between the 3 USPIOs (P > 0.05). At 2 hours p.i., SNR and DeltaSI data were significantly increased from baseline on T1-weighted images and significantly decreased on T2*-weighted images (P < 0.001). At 24 hours p.i., the T1-enhancement returned to baseline, whereas the T2*-enhancement remained significantly elevated (P < 0.001). Immunostains demonstrated an USPIO compartmentalization in macrophages in the arthritic synovium. CONCLUSIONS: Based on the relatively small number of animals in our study group, inflammation in antigen-induced arthritis can be equally detected and characterized with any of the three USPIOs evaluated. PMID- 16355041 TI - Controlling alcohol problems among hospitalized trauma patients. PMID- 16355042 TI - Interventions in trauma centers for substance use disorders: new insights on an old malady. PMID- 16355044 TI - Operational feasibility of interventions in trauma centers. PMID- 16355045 TI - Barriers to interventions for alcohol problems in trauma centers. AB - There is clear evidence that trauma-related serious injuries and fatalities can be directly linked to alcohol misuse. Because alcohol is a factor in many motor vehicle crashes (the leading cause of death for persons up to age 49), the National Highway Traffic and Safety Administration has set a high priority on decreasing alcohol-related motor vehicle crash injuries. Research now recognizes that most alcohol-impaired drivers who are injured are binge drinkers, not alcoholics. During the past decade, public health and substance treatment researchers have begun to focus on screening and early intervention in clinical settings as a means of reducing alcohol-related injuries. Trauma centers are uniquely positioned to address this problem because of the high percentage of alcohol-impaired patients admitted after motor vehicle crashes. This article discusses the barriers to implementing alcohol screening and intervention programs in trauma centers from the trauma surgeon's perspective. PMID- 16355046 TI - Implementing screening, brief intervention, and referral for alcohol and drug use: the trauma service perspective. AB - BACKGROUND: Most trauma surgeons are unfamiliar with screening, brief intervention, and referral (SBIR) programs for substance use disorders, and few trauma centers provide them. This report describes how an urban private-teaching hospital adapted a protocol from an existing emergency department-based program to include patients treated by the trauma service. METHODS: We recorded the rates of SBIR completion and reasons for failure during each phase of the implementation, interviewed trauma service staff and health educators to assess attitudes toward the program, and evaluated patient satisfaction surveys. RESULTS: By adding SBIR staff to the trauma outpatient clinic and to trauma morning rounds, the capture rate increased from 12 to 71%. Most screened patients (59%) were found at risk for problems or probably dependent on alcohol or drugs. Trauma service staff and health educators reported high satisfaction with the program. Patients reported higher satisfaction with SBIR. CONCLUSION: SBIR services can be effectively integrated into all components of a busy, urban trauma service by adding specially trained health educators to the trauma service staff. This collaboration provides effective SBIR services to both trauma and emergency service patients without interfering with patient flow or medical procedures. The relatively high percentage of patients at risk for alcohol or drug problems supports the inclusion of routine alcohol and drug screening for all eligible trauma patients. PMID- 16355047 TI - Feasibility of alcohol screening and brief intervention. AB - BACKGROUND: A variety of policy groups recommend that screening and brief intervention (SBI) programs for alcohol-use disorders be widely implemented in health care settings. This article reports the extent to which trauma surgeons support SBI programs and the feasibility of implementing these programs in trauma centers. METHODS: Trauma surgeons were surveyed to assess their support for implementing alcohol screening and brief intervention in trauma centers. To assess feasibility of implementation, three trauma centers implemented such programs. Each trauma center used one half-time research assistant who screened trauma inpatients for alcohol-use disorders and provided brief interventions for at-risk drinkers. The research assistant also recorded time spent screening and performing interventions, patient satisfaction with the intervention, and whether standard intervention elements were performed. RESULTS: Most surgeons surveyed supported alcohol screening, and 72% supported brief interventions. Research assistants who had no previous training in alcohol screening and brief interventions were successfully trained to screen and interview patients. One half-time research assistant was able to screen the eligible inpatient trauma population, with the exception of patients who were hospitalized on the weekends. Nearly 17% of patients at one trauma center were not screened because of language barriers. On any given day, roughly half the patients could not be screened because of the severity of their injuries. However, most of the patients were eventually screened during their hospital stay. Patient satisfaction was high. CONCLUSION: Most trauma surgeons supported alcohol screening and interventions. Preliminary data showed that one half-time research assistant at each facility could successfully screen most injured patients and implement brief interventions. An alcohol screening and brief intervention program seems feasible in any trauma center committed to implementation. PMID- 16355050 TI - Are we the problem? Overcoming obstacles to implementing intervention programs. AB - Alcohol-related injuries comprise a large percentage of injuries in the United States. As the impact of these injuries on society increases, a well-functioning trauma system becomes increasingly important. During the last decade, evidence based guidelines to reduce alcohol-related injuries have emerged. Further, evidence supports the effectiveness of brief intervention programs to reduce alcohol-related injuries and demonstrates that trauma centers can improve patient outcomes by integrating them into care. Although many obstacles have inhibited progress and made implementing preventive interventions a difficult task, economic constraints are among the biggest challenges to implementing intervention programs as part of routine trauma care. PMID- 16355051 TI - Confronting the obstacles to screening and interventions for alcohol problems in trauma centers. AB - Despite the demonstrated clinical benefits and decreased risks of injury recurrence, brief alcohol interventions are still not routine practice in trauma centers. Although alcohol and drugs play a significant role in trauma, few trauma specialists are aware of the potential benefits of interventions because alcohol treatment specialists have not widely disseminated their findings to other specialties. This article addresses some key obstacles that must be overcome to facilitate brief interventions as routine trauma practice. Included are discussions on training, cost and reimbursement factors, responsibility of the trauma surgeon, patient privacy and confidentiality issues, insurance laws and regulations, needed collaboration with partners, and research priorities and funding. PMID- 16355052 TI - Interventions-developing a plan for implementation. PMID- 16355055 TI - Alcohol interventions in trauma centers: the opportunity and the challenge. PMID- 16355056 TI - Brief motivational interventions: an introduction. AB - This article is an introduction to brief motivational interventions, which is an effective strategy to address alcohol-use disorders and the public health issues these disorders present. In this article, we summarize core concepts and our clinical experiences. To explore the contrast between these interventions and more traditional approaches to patient-provider interaction, the article describes strategies used in brief motivational interventions, answers common questions about the process, and provides references and resources for those who would like to learn more. PMID- 16355057 TI - The stages of change: when are trauma patients truly ready to change? AB - This article summarizes the Stages of Change model, which identifies five stages that people experience as they gradually move away from engaging in harmful behaviors to sustaining healthy behaviors. Patients in different stages of change need different kinds of interventions. The Stages of Change model enhances brief counseling interventions for trauma patients with substance use problems because counselors can now accurately choose an appropriate intervention strategy. The authors present three case studies illustrating the three earliest stages of change most commonly encountered in trauma center patients. PMID- 16355058 TI - The challenge of change. PMID- 16355059 TI - Changing the battle plan. PMID- 16355060 TI - Recommendations for trauma centers to improve screening, brief intervention, and referral to treatment for substance use disorders. PMID- 16355062 TI - The impact of alcohol and other drug problems on trauma care. PMID- 16355063 TI - Alcohol and trauma: the perfect storm. AB - Alcohol misuse, when combined with the right circumstances, culminates in a "Perfect Storm" that has catastrophic results. Alcohol misuse impairs judgment and increases the likelihood of serious injury. Once injured, the intoxicated patient is more likely to be hypotensive and less likely to be able to protect his or her airway. Alcohol also impairs multiple compensatory responses to injury that are critical to survival, thereby increasing the likelihood of serious complications. When complications do occur, they may be more severe for intoxicated patients because both acute and chronic ethanol use adversely affect immunity. Thus, all phases of trauma care are potentially affected by excessive alcohol use. This paper presents facts about trauma care and the physiologic consequences and clinical implications of alcohol intoxication. Further, it shows how health-care costs increase when evaluating and managing an intoxicated trauma patient. This financial burden further escalates in the surgical ICU because the care of an intoxicated patient falls outside the realm of standard care. Trauma surgeons, perhaps more than other health-care providers, have a unique opportunity and a responsibility to address potential alcohol misuse with their patients. They witness the Perfect Storm almost daily and are fully aware of the short- and long-term consequences of alcohol misuse. A trauma center visit provides an opportune time and place to incorporate alcohol screening and brief interventions as a part of standard trauma care. PMID- 16355064 TI - The impact of street drugs on trauma care. AB - The use of illicit drugs, specifically heroin and cocaine, complicates trauma patient management and consumes extensive hospital resources. This paper focuses on heroin- and cocaine-related injuries observed by physicians at Detroit Receiving Hospital, a large urban Level I trauma center. The pharmaceutical effects, mode of administration, and the manner in which these drugs affect diagnosis and treatment of injuries are documented and discussed. Specific drug related complications associated with overdose, soft-tissue infections, bacterial endocarditis (therapy resistant), vascular thromboses, vascular aneurysms, vasoconstriction, stroke, cardiac arrhythmias, muscle ischemia, and solid-organ abscesses are also analyzed. Illicit drug use significantly complicates initial diagnosis and trauma management and is associated with severe adverse pathophysiologic effects. Currently, prevention efforts, such as interventions in trauma centers, should be considered as the most efficient and feasible way to prevent injury recidivism in this patient population. We also conclude that legislative change may be the answer in reducing or preventing the horrendous problems caused by illicit drugs. PMID- 16355065 TI - Alcohol Problems among Hospitalized Trauma Patients: Controlling Complications, Mortality, and Trauma Recidivism. May 28-30, 2003, Arlington, Virginia, USA. Conference proceedings. PMID- 16355066 TI - A rational approach to formulating public policy on substance abuse. AB - Unlike alcohol, which is legal and regulated, current public policy makes drugs such as heroin, cocaine, methamphetamines, and marijuana illegal. This article summarizes the history of drug and alcohol use in the United States, compares our public policies on alcohol to those on drugs, and shows the direct link between alcohol or drug use and crime, corruption, violence, and health problems in other countries and in our own. A rational approach to formulating a workable public policy is presented. PMID- 16355070 TI - Session 2: substance-abuse interventions-setting the stage for discussion. PMID- 16355071 TI - Screening and interventions for alcohol and drug problems in medical settings: what works? AB - This article summarizes current knowledge about the accuracy of screening tests and the efficacy of interventions for substance use disorders in different medical settings (including trauma centers) where the practitioners are not specialists in the management of substance use disorders. In the first section, we introduce basic screening approaches for psychoactive substance use disorders and issues of natural history, risk factors, and populations at risk. Next, we review recent scientific research on the development of screening tests and the evaluation of early intervention services for persons at risk. We conclude that reliable and valid screening tests are available to detect alcohol use disorders but that further work is needed before routine screening for drug use disorders is warranted. We found strong evidence to support the effectiveness of brief interventions in managing at-risk drinkers; however, the evidence is only suggestive for drug use disorders. Finally, we explore the implications of the findings for developing a public health approach to early intervention, particularly as it relates to the unique needs of trauma centers. PMID- 16355072 TI - Brief interventions for hospitalized trauma patients. AB - Substance abuse is one of the most prevalent comorbid conditions among trauma patients. Research has shown that injury can be reduced when brief interventions are provided to trauma patients who are hospitalized for substance abuse. This article presents data from a Level I trauma center that provided brief interventions daily over a 5-year period. A generic model for brief intervention is described, along with concrete details of this addiction intervention service. PMID- 16355074 TI - Why does knee extensor muscles torque decrease after eccentric-type exercise? AB - AIM: The purpose of this study was to re-examine central and peripheral origins of neuromuscular fatigue after a highly strenuous eccentric exercise of the knee extensor muscles (KE) using both voluntary/evoked contractions and electromyographic recordings (EMG). METHODS: Before, and 30 min after 15 min of intermittent one-logged downhill running, maximal percutaneous electrical stimulations (single twitch, 0.5 s tetanus at 20 Hz and 80 Hz) were applied to the femoral nerve of 10 male subjects. Electrically evoked superimposed twitches were delivered during isometric maximal voluntary contraction (MVC) to determine maximal voluntary activation (%VA). Vastus lateralis (VL), vastus medialis (VM) and biceps femoris (BF) EMG were recorded during MVC and quantified using the root mean square (RMS) value. M-wave characteristics were also determined. RESULTS: KE MVC and %VA decreased significantly with fatigue (-19.6+/-6.1%; P<0.001 and -7.8+/-6.6%; P<0.01, respectively). Peak tetanus tension at 20 and 80 Hz (P20 and P80, respectively) declined (P<0.001), concurrently with a decrement of the P20 x P80(-1) ratio (-37.3+/-16.6%; P<0.001). Antagonist muscle coactivation, RMS to M-wave peak-to-peak amplitude and MVC x P80(-1) ratios were unchanged after the fatiguing exercise. CONCLUSIONS: The results reveal that part of the large loss in MVC may have a central origin but most of the MVC decrement is due to the presence of low-frequency fatigue while possible contractile failure cannot be excluded. PMID- 16355075 TI - Effects of anthropometric parameters and stride frequency on estimation of energy cost of walking. AB - AIM: This study was conducted to develop a generalized, multifactorial walking equation for estimating VO2 during treadmill walking at self-selected treadmill belt speeds. Also, the study sought to find the stride frequency (SF) and stride length (SL) combination which yielded the highest VO2. METHODS: The participants for this study included 100 university students (50 male and 50 female) between the ages of 18 and 57. After standing height, sitting height, hip breadth, calf length, thigh length, and gross body mass were measured, the preferred speed (Pv), self-selected as being challenging but comfortable, was determined for each subject at 0% grade. Subsequently, a 2nd walk test was conducted at Pv utilizing 5 different SF which were paced by means of a metronome. In each of these 5 SF-SL combinations, heart rate and metabolic data were recorded. RESULTS: Stepwise regression analysis showed that calf length and treadmill belt speed were two independent variables that account for significant increase in variance in which VO2 at preferred stride Frequency (PSF)-15 strides was used as dependent variable. Split plot ANOVA results indicated that the participants' VO2, heart rate, and breathing frequency responses were significantly (p<0.05) lower at PSF when compared to the other 4 SF conditions. CONCLUSIONS: Research indicated that other than velocity of walking, anthropometric characteristic such as calf length has a significant effect on estimation of energy cost of walking. Furthermore, cardiovascular loading can be affected by manipulating SL and SF as alternate forms of training in regimens of walking. PMID- 16355076 TI - Ground reaction forces and heart rate profile of aerobic dance instructors during a low and high impact exercise programme. AB - AIM: The magnitude of ground reaction forces (GRF) has been associated, although never verified, with the high incidence of lower extremities injuries in aerobic dance (AD) instructors. Moreover, during their working activities AD instructors have demonstrated HR levels, such as 70% HRmax, values, more in training rather in working status. This study was designed to investigate GRF and heart rate (HR) exhibited by AD instructors of both genders, during a simulated AD instruction, from the perspective of accepted occupational workloads. METHODS: Fourteen females and 14 males instructors performed a 35 min AD exercise programme (warm up--low impact (LI) interval--in high impact (HI) interval--cool down). Four GRF measurements were taken during LI and HI time intervals, respectively. HR was recorded throughout the whole experimental procedure and was synchronised to GRF measurements. RESULTS: All GRF and HR values were significantly increased in HI exercise (p<0.05) with a non significant (p>0.05) time effect for GRF. In both LI and HI exercises, females demonstrated significantly higher vertical but lower lateral GRF (p<0.05) and significantly shorter cycles of movement (p<0.05) while in HI exercise they had significant longer flight times (p<0.05). For both genders, HR was kept at 70% and 80% of HR(max-calc) and RHR was 60% and 70%, during LI and HI intervals respectively, with females showing a trend, though non significant, for higher HR values. CONCLUSIONS: The gender specificity of the significant vertical and lateral GRF pattern differences, may possibly be associated with the significant anthropometric differences of male and female AD instructors. HR(max-calc) and RHR exceeded the accepted occupational levels rising to training status levels. We suggest that AD instructors take up such instructing methods which allow them to minimize the magnitude or the rate of GRF, as well as HR levels, developed in the course of their working hour. PMID- 16355077 TI - Indoor physical education measured by heart rate monitor. A case study in Portugal. AB - AIM: Reports indicated that children are spending too much time being physically inactive during physical education classes and many school physical education programs are not meeting the recommendation. The purpose of this study was to investigate the physical activity levels in Portuguese indoor physical education classes. METHODS: The physical activity levels of 28 Portuguese 7th grade children were assessed by heart rate monitor during their indoor physical education classes. RESULTS: The study showed that much physical education class time had been wasted, and less than 70% of physical education class time had actually been used in classes. On average, 14.4 minutes of 45-min physical education classes and 27.9 min of 90-min physical education classes had been spent in MVPA with children's heart rates above 139 bpm; 6.7 min of 45-min physical education classes and 15.7 min of 90-min physical education classes had been spent in VPA with children's heart rates above 159 bpm. Children engaged in more MVPA, VPA, and continuous bouts of physical activity in 90-min physical education classes than those of 45-min physical education classes. There were no significant differences in activity levels between genders in co-educational indoor physical education classes. CONCLUSIONS: This indoor PE class study supports the results that children are less active during school physical education classes. Both the quantity and the quality of school physical education needs to improve to encourage children to engage in a substantial amount of physical activity both inside and outside school. PMID- 16355078 TI - Effects of Panax ginseng extract on exercise-induced oxidative stress. AB - AIM: This study was designed to study the effects of Panax ginseng extract (PGE) on lipid peroxidation and scavenger enzymes induced by an acute exhaustive exercise in sedentary humans. METHODS: Seven healthy male subjects performed 2 exhaustive incremental exercises on the treadmill before and after 8 weeks' PGE ingestion (2 g each time, 3 times a day) as the control and PGE exercise, respectively. VO2, HR, and exercise duration during exercise were measured. Blood samples were collected at rest, and immediately, 10 and 30 min after each test and used to measure malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD). RESULTS: PGE administration significantly increased exercise duration until exhaustion by 1.5 min (p<0.05). MDA was significantly elevated following both trials (p<0.01), however, it was attenuated after PGE administration (p<0.01). CAT and SOD activities following exercise were significantly elevated, but the activities following control exercise were much lower than those following PGE exercise. CONCLUSIONS: These results suggest that the elevation in CAT and SOD activities as scavenger enzymes after PGE administration result in decrease of MDA level as one of PGE action mechanisms and consequently, prolong exercise duration until exhaustion. These findings support scientific claims that ginseng has ergogenic properties in facilitating recovery from exhaustive exercise. PMID- 16355079 TI - The effects of proprioceptive neuromuscular facilitation stretching on shoulder range of motion in overhand athletes. AB - AIM: The purpose of this study was to compare the effects of the contract-relax contract (CRC) and hold-relax-contract (HRC) proprioceptive neuromuscular facilitation (PNF) stretching programs against a control, on external range of motion (ROM) of the shoulder in apparently healthy athletes. METHODS: The subjects were male and female adults between the ages of 25 to 50 years old. Subjects also had been involved in at least one overhand throwing sport (tennis, baseball, quarterback in football, etc.) in the past year. There were 30 participants whom were randomly assigned to 1 of 3 groups (CRC PNF, HRC PNF, control) with 10 subjects per group. Measurement of ROM for external rotation of the shoulder was performed prior to and after 6 weeks of training using a goniometer. The CRC and HRC PNF stretches were performed on subjects twice a week for 6 weeks. The statistical analysis conducted involved a 3x2 analysis of variance with the criteria for statistical significance set at p<0.05. Post hoc testing included paired t-tests and Tukey tests to pinpoint significant differences relative to the interaction between group and time. RESULTS: There was an increase in ROM from pretest to post-test for the HRC group (+13.50 degrees) and CRC group (+14.60 degrees), but not in the control group (+0.30 degrees). The improvement in ROM however, was similar between the HRC and CRC groups. CONCLUSIONS: The CRC and HRC PNF stretching techniques are effective at increasing external shoulder ROM when consistently performed 2 times a week for 6 weeks. PMID- 16355080 TI - Quantification of tension in Thera-Band and Cando tubing at different strains and starting lengths. AB - AIM: Elastic resistance exercise is frequently used in rehabilitation programs. Precise quantification using color coded bands is not possible. A method to predict forces for any length of mostly used elastic tubings and thereby a general equation for the comparison of elastic material is presented. METHODS: In an in vitro-series the tubing tension in relation to elongation of 6 colors of Thera-Band and Cando tubings was directly measured. The relation between tension and elongation was determined depending on different resting lengths. In clinical practice tension and elongation are prescribed. Two mathematical methods to calculate the resting length as regulatory variable are presented. One is based on the construction of iso-tension curves, the 2nd on the general mathematical relation delta % tension=K* % length change+n. RESULTS: 1. The tension/elongation relation is linear beyond a threshold in the early part of elongation. 2. The iso tension method works graphically and facilitates the precise determination of resting length. 3. The double % relation between tension and elongation method describes the general properties of all tubings, where K=0.75 is a general constant. To calculate absolute values for the resting length, corresponding to a given tension and elongation, special ''color constants'' are presented. CONCLUSIONS: The prediction of maximum tension values for any given elongation is possible by simple variation of resting length. These findings may improve the usefulness of elastic tubings in rehabilitation programs. The mathematical method further may be used for the comparison of elastic materials with linear characteristics. PMID- 16355081 TI - Fitness centre membership is related to healthy behaviours. AB - AIM: Maximizing the health benefits associated with reducing inactivity levels requires an understanding of the individual and environmental determinants of physical activity. Membership in a fitness facility promotes physical activity, yet little is known of its relationship to health. The purpose of this study was to compare physical activity levels, and health status, behaviours, and beliefs, in members of a fitness centre, and non-member community residents. METHODS: Using a cross-sectional design, fitness centre members (n=236) and non-members (n=302) were compared with respect to perceived health status, use of health care services, fitness status, physical activity level, perceived control over health, and the likelihood of engaging in health promoting behaviours, using The Health Promoting Lifestyle Profile. Questionnaires were mailed to adult members of a fitness centre, and a stratified (age, sex) sample of non-members randomly selected from the local community. RESULTS: Fitness centre members were more likely than the comparison group to have visited a general physician, dentist, athletic therapist, optometrist, or nutritionist during the previous year (p<0.05), to exercise regularly, and to rate their physical fitness as very fit. They scored significantly higher on the overall health promoting lifestyle score (p=0.0353) as well as on health responsibility (p=0.0053), exercise (p=0.0001), and nutrition (p=0.0166) subscales, even after adjusting for differences in activity levels between groups. CONCLUSIONS: Fitness centre membership is associated with increased health responsibility and health promoting behaviours. This finding appears to be related to membership in the fitness centre, and not to increased participation in physical activity. PMID- 16355082 TI - Personality does not influence exercise-induced mood enhancement among female exercisers. AB - AIM: The present study investigated the influence of personality on exercise induced mood changes. It was hypothesised that a) exercise would be associated with significant mood enhancement across all personality types, b) extroversion would be associated with positive mood and neuroticism with negative mood both pre- and post-exercise, and c) personality measures would interact with exercise induced mood changes. METHODS: Participants were 90 female exercisers (M=25.8 y, SD=9.0 y) who completed the Eysenck Personality Inventory once and the Brunel Mood Scale before and after a 60-min exercise session. Median splits were used to group participants into 4 personality types: stable introverts (n=25), stable extroverts (n=20), neurotic introverts (n=26), and neurotic extroverts (n=19). RESULTS: Repeated measures MANOVA showed significant mood enhancement following exercise across all personality types. Neuroticism was associated with negative mood scores pre- and post-exercise but the effect of extroversion on reported mood was relatively weak. There was no significant interaction effect between exercise-induced mood enhancement and personality. CONCLUSIONS: Findings lend support to the notion that exercise is associated with improved mood. However, findings show that personality did not influence this effect, although neuroticism was associated with negative mood. PMID- 16355083 TI - The influence of intrinsic motivation on an endurance field test. AB - AIM: The aim of the present study was to examine the influence of various aspects of intrinsic motivation on the levels of performance in an endurance field test. METHODS: The sample of the study consisted of 144 undergraduate Physical Education students. A Greek version of the Intrinsic Motivation Inventory (IMI) was administered to participants after the completion of the 20 m shuttle field test. RESULTS: The 20 m shuttle run test was significantly and positively associated with the interest-enjoyment, perceived competence and effort importance subscales. Regression analysis showed that only the perceived competence subscale could significantly contribute to the prediction the 20 m shuttle run scores. CONCLUSIONS: It was concluded that the perceived competence subscale of the IMI significantly predicted 20 m shuttle run performance and thus could be used to explain an additional percent of the variation in the participants' performance scores. PMID- 16355084 TI - Lifelong physical training prevents the age-related impairment of heart rate variability and exercise capacity in elderly people. AB - AIM: Aging is associated with a reduction on heart rate variability (HRV) and working capacity. Aim of this study was to evaluate in a group of elite master athletes the effect of a lifelong history of endurance running on HRV and exercise working capacity. METHODS: Twenty athletes (males, age 68.5+/-4.5 years) who practiced endurance running for at least 40 years, and 20 age-sex-matched control subjects with sedentary lifestyle were studied. All the participants underwent a maximal stepwise electrocardiogram (ECG) on effort (work-rate increments of 25 Watts every 2 min) and a 24-hour ECG monitoring. RESULTS: All the time domain measures of HRV and the LF and HF powers were significantly higher in elderly athletes than in sedentary subjects (P<0.001), while the LF/HF ratio was comparable between the 2 groups. Athletes exhibited significantly higher workload than controls (1610+/-489 vs 687+/-236 W, P<0.0001). Both the groups achieved, at maximum workload, similar heart rate (142+/-10 vs 138+/-18 bpm, ns), systolic blood pressure (226+/-18 vs 220+/-16 mmHg, ns), and rate pressure product (32,596+/-2952 vs 30,838+/-3675, ns). Maximum work-rate attained in athletes was 225 W. By contrast, none of the controls reached a work-rate higher than 150 W. In the whole group we also showed a positive correlation between the time domain HRV parameter SDNN and maximum workload (r=0.58, P<0.001). CONCLUSION: Long-term endurance training induces in elderly subjects an increased HRV and a higher exercise working capacity, which are well-established predictors of cardiovascular and overall mortality. PMID- 16355085 TI - Treadmill running promotes functional recovery and decreases brain-derived neurotrophic factor mRNA expression following sciatic crushed nerve injury in rats. AB - AIM: Peripheral nerve injuries are commonly encountered clinical problems and often result in severe functional deficits. In the present study, the effects of treadmill running on the recovery rate of locomotor function and the expression of brain-derived neurotrophic factor (BDNF) mRNA following sciatic crushed nerve injury in rats were investigated. METHODS: EXPERIMENTAL DESIGN: Comparative investigation was performed over 14 days. SETTING: Experimental animal laboratory. PARTICIPANTS: 24 male Sprague-Dawley rats weighing 200+/-10 g. Animals were randomly assigned into 3 groups: the sham-operation group, the sciatic nerve injury group, and the sciatic nerve injury and running group. INTERVENTIONS: The right sciatic nerve was crushed for 30 s using a surgical clip. Rats of the running group were made to run on a treadmill for 30 min once a day for 12 consecutive days. MEASURES: Functional recovery was analyzed using a walking track analysis which can be quantified with the sciatic function index (SFI) and BDNF mRNA expression was analyzed using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Sciatic crushed nerve injury showed characteristic gait changes showing decrease of SFI value and treadmill running significantly enhanced the SFI value. The level of BDNF mRNA expression was increased following sciatic crushed nerve injury and treadmill running significantly suppressed the sciatic nerve injury-induced increment of BDNF mRNA expression. CONCLUSIONS: It can be suggested that treadmill running after peripheral nerve injury is effective in the functional recovery by inhibition on the over-expression of BDNF mRNA. PMID- 16355086 TI - Examination of quantitative and fractal analysis of sway characteristics of the center of foot pressure movement during a static upright posture. Analysis based on alcohol intake. AB - AIM: This study aimed to examine the sway characteristics of center of foot pressure (CFP) movement during a static upright posture under the influence of alcohol by using quantitative and fractal analysis. METHODS: Eleven healthy young people participated. They drank in a range of 0.54-1.83 ml/kg of alcohol, standardized by body mass, within 10 min. Blood pressure, heart rate, 2 types of nervous function tests and the CFP movement were measured before and after the alcohol intake. Thirty-six parameters of the CFP movement were used for quantitative analysis. Diffusion coefficient, scaling exponent and critical point coordinates were used in fractal analysis. RESULTS: Quantitative analysis confirmed that the CFP parameters such as distance, velocity and area were significantly changed, although the parameters evaluating cyclical characteristics and center average were not. Fractal analysis confirmed that critical point coordinates existed at time-lag=1.0 before and after the alcohol intake, and the short-term region (ST) and long-term region (LT) could be separated. A diffusion coefficient in the ST was larger than that in the LT, and the scaling exponent was over 0.5 in the ST and under 0.5 in the LT. CONCLUSIONS: Influences of alcohol intake were found in amount of body-sway, but not in the displacement or periodicity of the CFP parameters. Fractal analysis confirmed that CFP movement was dependent on time-series, and its characteristics changed before and after the time-series critical point and was unsteady. PMID- 16355087 TI - Angioplasty and stenting of extracranial vertebral artery stenosis. AB - Percutaneous angioplasty and stent placement seem a useful technique for the treatment of vertebro-basilar insufficiency and the first treatment to be proposed. This technique appears safe and effective for alleviating symptoms and improving blood flow to the cerebral circulation, with a low complications rate and good long-term results. However, this procedure needs experienced interventionists to choose the stent and have appropriate placement of the stent in the ostium of the vertebral artery (VA). The tortuosity of the VA may be technically challenging. The new coronary stents seem to be well suited to treat atherosclerotic lesions of the origin and of the proximal VA. A large variability of restenosis risk has been reported. Drug eluting stents may be the solution. Prospective randomized studies are needed to demonstrate the clinical effectiveness of VA stenting in stroke prevention, its durability, and to define more clearly its indications. PMID- 16355088 TI - Effects of elastic stocking on quality of life of patients with chronic venous insufficiency. An Italian pilot study on Triveneto Region. AB - AIM: Even though compression therapy is the most recommended treatment for chronic venous insufficiency (CVI) in the national and international guidelines, its application, at least in Italy, is lower than the estimated need from the prevalence of CVI in the epidemiological studies. Since we believe that the measurement of the impact of compression therapy on quality of life (QoL) could improve the compliance for this precious treatment, we carried out this study on 50 patients with CVI. METHODS: Fifty patients (23 CEAP C2 and 27 C(3-4-5), selected within a larger study on QoL in CVI), received a prescription for compression therapy. Before treatment and 4 months afterwards, the patients received the instrument for QoL assessment (MOS SF-36; CIVIQ-2; Euro-QoL 5D and a visual analogue scale). The score scales have been adjusted to poorest QoL as 0 and best QoL as 100. RESULTS: Baseline QoL of patients in class C2 did not show significant difference with the healthy Italian Population, except for the physical role item. The patients in class C(3-4-5) showed significant reduction of QoL. After 4 months all the items of the utilized instrument showed significant improvement (from P<0.01 to P<0.04 for SF-36; from P<0.099 to P<0.006 for other instruments) in all CEAP classes. CONCLUSIONS: The study unquestionably shows that the compression therapy improves the QoL of patients with CVI, and should be included in the CVI treatment covered by the Public National Health Insurance. Finally, the QoL measurement could be utilized as the scientific method to assess the effectiveness and efficacy of different therapeutic devices. PMID- 16355089 TI - Comparison of two intermittent pneumatic compression systems. A hemodynamic study. AB - AIM: Sequential leg compression has been previously shown to be superior to uniform compression. The aim of our study was to compare the hemodynamic effectiveness of the portable sequential compression device (SCD Express Compression System, Tyco Healthcare Group LP, Mansfield, MA, USA) with a rapid inflation device (VenaFlow, Aircast, Inc, Summit, NJ, USA). The former, by sensing venous refill time, commences compression when the calf veins are refilled. METHODS: The two devices were tested in 12 normal volunteers in the semirecumbent position using duplex ultrasound. Baseline and augmented flow velocity and volume flow were measured at the level of the common femoral vein, above the saphenofemoral junction. Refilling time was determined from velocity recordings of the common femoral vein. Total and peak volume of blood expelled per hour during compression were calculated using flow data and the individual cycling rate. RESULTS: Both devices increased venous flow velocity, up to 3.8 times the baseline (all P<0.001). Refill time of the rapid inflation device was shorter in comparison with the sequential compression device (15+/-2.2 vs 25+/-4 s; P<0.001), suggesting incomplete vein evacuation. The sequential compression device, by augmenting flow throughout a significantly longer compression period per cycle (10.9 s vs 6.3 s), expelled significantly more venous blood (121+/-68 vs 81+/-63 mL; P<0.001). Similarly, the total volume of blood expelled per hour with the sequential compression device was 100% higher than the rapid inflation device (9685+/-5426 vs 4853+/-3658 mL; P<0.001). Although peak velocity enhancement was higher with the rapid inflation device, flow augmentation (a product of average blood flow velocity) was comparable (669+/-367 vs 771+/-574 cm/s; P=0.223) with the sequential compression device, mainly because the rapid inflation device failed to maintain flow enhancement beyond the initial flow surge. CONCLUSIONS: Sequential compression showed hemodynamic superiority compared to a rapid inflation device. This was enhanced further by the sensing of refill time, which resulted in more compression cycles over time. The relative efficacy of the two devices in deep vein thrombosis prevention should be tested in future studies. PMID- 16355090 TI - Impaired ambulatory venous function in lymphedema assessed by near-infrared spectroscopy. AB - AIM: The aim of this study was to investigate venous function in patients with leg lymphedema during exercise using near-infrared spectroscopy (NIRS), compared with that of patients with chronic venous insufficiency (CVI). METHODS: Forty three legs of 33 patients (5 males, 28 females; mean age: 53 years) with leg lymphedema without varicose veins or deep vein thrombosis underwent a treadmill walking test with simultaneous NIRS. For comparison, 136 legs of 91 patients (35 males, 56 females; mean age: 56 years) with varicose veins as a CVI group and 45 legs of 38 healthy volunteers (23 males, 15 females; mean age: 50 years) were also evaluated in the same method. Deoxygenated hemoglobin (HHb) was continuously measured during exercise, and the ambulatory venous retention index (AVRI) of each leg was obtained from the serial changes in HHb. RESULTS: The mean AVRI of the lymphedema group was significantly higher than that of healthy legs and significantly lower than of legs with moderate or severe CVI. Furthermore, it was similar to that in the mild CVI group. CONCLUSIONS: Venous function is impaired in exercising legs with lymphedema, and corresponds to that in legs with mild venous insufficiency. The treatment of lymphedema should take CVI into consideration. PMID- 16355091 TI - Long-term results of combined iliac endovascular intervention and infrainguinal surgical revascularization for treatment of multilevel arterial occlusive disease. AB - AIM: The purpose of this study was to investigate the long-term results of combined iliac endovascular intervention and infrainguinal surgical revascularization. METHODS: A retrospective review of 39 infrainguinal bypasses combined with iliac endovascular intervention in 35 patients over a 16-year period was performed (Combined group). These results were compared to those of 43 infrainguinal bypasses performed with suprainguinal bypass operation in 39 patients (Surgical group), who had iliac lesions not amenable to angioplasty/stenting. There was no significant difference in the preoperative limb ischemic symptoms between them. RESULTS: Although the proportion of patients with coronary artery disease in the Combined group was significantly higher than that in the Surgical group, there was a trend toward lower morbidity/mortality in the Combined group compared with the Surgical group (8.6% vs 15.4%; P=0.3706). No significant differences in the rates of clinical and hemodynamic improvement and limb salvage rate were observed between the two groups. The primary patency rate of infrainguinal bypass at 1, 3, and 5 years was 83.2%, 80%, and 71.2% in the Combined group, and 97.1%, 89.9%, and 80.5% in the Surgical group, respectively. The secondary patency rate at 1, 3, and 5 years was 91.9%, 91.9%, and 76.3% in the Combined group, and 97.1%, 89.9%, and 84.6% in the Surgical group, respectively. Importantly, there was no significant difference in the primary/secondary patency rates between the two groups (Kaplan-Meier, log-rank test, primary patency, P=0.116; secondary patency, P=0.4407). CONCLUSIONS: Infrainguinal surgical reconstruction combined with iliac endovascular procedure may reduce operative risk, and further, long-term patency is comparable to that in the Surgical group. PMID- 16355092 TI - The impact of patient characteristics on long-term results of above-knee prosthetic femoropopliteal bypass for critical ischemia. AB - AIM: The aim of this paper was to study the impact of risk factors on long-term results of above-knee prosthetic femoropopliteal bypass for critical ischemia. METHODS: One hundred and eleven consecutive operations (108 patients) were done between January 1990 and December 2001. All cases were prospectively registered. The patient characteristics were subjected to univariate analysis using the log rank test for impact on survival, limb salvage and patency rates. Variables approaching significance (P<0.1) were included in multivariate analyses performed with the Cox proportional hazard model. RESULTS: The 30-day mortality rate was 5.5%. The 2- and 5-year survival was 72% and 42%, respectively. Twenty-seven limbs were subjected to major amputations during follow-up. The limb salvage rates at 2 and 5 years were 83% and 73%. The 2- and 5-year assisted primary patency rates were 45% and 17% versus 52% and 27% for the secondary patency. The 2-year primary patency rate for smokers was 38% versus 62% for non-smokers (P=0.018, hazard ratio 2.18). Smoking and tissue loss were significantly associated with reduced secondary patency rates on multivariate analysis. CONCLUSIONS: The inferior primary patency rates of smokers indicate that prosthetic femoropopliteal bypass for critical leg ischemia should not be the primary treatment option for these patients. The poor secondary patency rates for smokers as well as for patients with tissue loss suggest that these patients may benefit from alternative treatment modalities, instead of reopening an occluded bypass. PMID- 16355093 TI - A comparison of the inflammatory response and the recovery of bowel function between trans- and extraperitoneal approaches of abdominal aortic aneurysmectomy. AB - AIM: To determine whether surgical stress is less with transperitoneal or extraperitoneal abdominal aortic aneurysmectomy, blood concentrations of inflammatory cytokines and other inflammatory markers with recovery of bowel function were examined. METHODS: Patients who underwent abdominal aortic aneurysmectomy electively via the transperitoneal (T-group; n=15) or the extraperitoneal approach (E-group; n=17) were evaluated. Inflammatory cytokines (interleukin[IL]) IL-6, IL-8, C-reactive protein concentrations, and systemic inflammatory response syndrome score were determined before operation and after operation on days (POD) 1, 3, 7, and 14. Recovery of bowel function was estimated by the time taken for resumption of bowel movement and oral intake, and by the volume of fluid collection from the nasogastric tube. RESULTS: Cytokine (IL-8) concentrations were higher in the T-group than the E-group with significant difference on POD 7 (4.8+/-0.5 versus 3.4+/-0.2 pg/mL, respectively; P=0.02). Recovery of bowel function and oral intake were earlier, and the volume of fluid collection from the nasogastric tube was smaller in the E-group than the T-group significantly. CONCLUSIONS: Early recovery of bowel function and low concentration of inflammatory cytokines suggest that the extraperitoneal approach to the abdominal aorta is less stressful to the transperitoneal approach. Postoperative inflammatory response may mainly depend on damage of the bowel in the operation of the abdominal aortic aneurysm. PMID- 16355094 TI - Data density curves of B-mode ultrasound arterial wall thickness measurements in unaffected control and at-risk populations. AB - AIM: Atherosclerosis is a slow disease process of arterial walls with onset decades prior to its clinical manifestations. Lifelong follow-up data may help to identify and understand the pathophysiology of this process. These longitudinal data are scarce. Using a standardized imaging and image analysis protocol, we acquired cross-sectional data of carotid and femoral arterial wall segments in populations at different cardiovascular disease risk. METHODS: B-mode ultrasound intima-media thickness (IMT) data of carotid and femoral arteries were acquired in individuals at high cardiovascular disease risk: 44 young adolescents with familial hypercholesterolemia (FH), 248 adult FH patients and 184 patients with coronary artery disease (CAD), as well as in disease free unaffected individuals, 44 young adolescents, 26 middle-aged adults and 48 senior adults. RESULTS: Per patient combined average IMT (SD) and % of lesions in the high risk populations were 0.55 (0.05) mm, 0.1%, 0.86 (0.18) mm, 15%, and 0.9 (0.18) mm, 18%, respectively. In the unaffected groups these values were 0.53 (0.03) mm, 0%, 0.59 (0.07) mm, 0%, and 0.77 (0.12) mm, 8%. Of all arterial segments, the far wall of the common femoral artery (CFA) of the FH patients exhibited the highest absolute IMT (1.12 [0.61] mm), the most rapid estimated IMT increase since adolescence (+0.58 mm) and the highest percentage of lesions (39% of CFA measurements). CONCLUSIONS: Regardless of location, carotid and femoral arterial walls increase in thickness with age and cardiovascular disease risk. This increase in thickness and prevalence of lesions is not similarly distributed among anatomical segments. The strong preponderance in arterial wall segments with the highest estimated atherosclerosis progression indicates the existence of a threshold value beyond which plaque formation is greatly increased. In the set of arterial locations we studied, this process might be best represented by the far wall of the CFA. PMID- 16355095 TI - TFPI antigen and activity levels in patients with asymptomatic atherosclerosis and target organ acute and chronic complications. AB - AIM: In patients with atherosclerosis there is an activation of the tissue factor mediated coagulation cascade. The aim of our study was to ascertain if there is a relationship between increased tissue factor pathway inhibitor (TFPI) antigen and activity plasma levels and atherosclerosis. METHODS: DESIGN: case-control study. SETTING: Department of Internal Medicine and Cardiovascular Diseases, University of Palermo and Laboratory of Genetics and Molecular Biology, Mayo Clinic, Rochester, MN, USA. PATIENTS: 63 consecutive patients with asymptomatic atherosclerosis or with its acute or chronic complications, and 20 healthy volunteers. MEASUREMENTS: TFPI antigen was detected by an immunoenzimatic assay (Imunobind total TFPI ELISA kit, American Diagnostica Inc., USA). TFPI activity was evaluated by an activity assay (Actichrome TFPI activity assay, American Diagnostica Inc., USA). RESULTS: PATIENTS with chronic (P=0.0001 for TFPI Ag, P=0.006 for TFPI Ac) and acute (P=0.04 for TFPI Ag, P=0.01 for TFPI Ac) vascular disease and with asymptomatic carotid plaque (P=0.0019 for TFPI Ag, P<0.05 for TFPI Ac) had significantly increased TFPI antigen and activity plasma levels vs healthy volunteers. Moreover, patients with chronic vascular disease had higher TFPI Ag levels vs patients with asymptomatic atherosclerosis (P=0.0013). CONCLUSIONS: The originality of our study was the finding of increased TFPI levels, not only in patients with acute complication of atherosclerosis, but also in those with chronic vascular disease and with asymptomatic carotid plaque. PMID- 16355096 TI - Angiocidal effect of Cyclosporin A: a new therapeutic approach for pathogenic angiogenesis. AB - AIM: Angiogenesis is essential in the development of several disorders such as cancer, arthritis, and autoimmune diseases. Several agents prevent angiogenesis but only a few destroy established angiogenesis. In this study we tested whether local or systemic administration of Cyclosporin A (CyA) would inhibit as well as destroy established angiogenesis in an in vivo assay of angiogenesis. METHODS: We utilized an in vivo assay of angiogenesis in which an angiogenic mixture of Matrigel, FGF, VEGF, and heparin was injected subcutaneously into mice. Angiogenesis in the subcutaneous plugs was quantified by ANOVA. CyA or the vehicle for CyA was administered to the experimental or the control groups by three routes: by addition to the angiogenic mixture, by local injection into the angiogenic plug at various time points or by systemic administration at high doses. Angiogenesis was quantified by pointing method and expressed as an angiogenic index (AI). RESULTS: In control animals the subcutaneous plug of Matrigel with the angiogenic mixture revealed exuberant angiogenesis at day 4 and day 7. This angiogenesis was completely inhibited when CyA was included in the angiogenic mixture; the vehicle for CyA had no such effect. Angiogenesis that had progressed was found to regress after local subcutaneous injection of CyA at day 4 and 7. Similar regression of angiogenesis was noted when CyA was administered systemically after allowing angiogenesis to proceed for 4 days. CONCLUSIONS: Our experiments strongly suggest that CyA is both angiocidal and angiostatic in vivo. These results provide a basis for future therapy directed against established angiogenesis in malignancies and autoimmune diseases. PMID- 16355097 TI - Preoperative measurement of the intrathrombotic pressure of a thrombosed popliteal artery aneurysm for decision of surgical indication. AB - To evaluate the risk of rupture of a 77 mm thrombosed popliteal artery aneurysm (PAA) in an 82-year-old woman, the aneurysm was percutaneously punctured and the intrathrombotic pressure (ITP) was measured preoperatively. The percentage of the mean ITP to the mean brachial artery pressure was 80%. The aneurysm was judged to have the risk of rupture, and replacement with a prosthetic graft was performed. The percentages of the mean ITP to the mean radial artery pressure and to the mean femoral artery pressure measured intraoperatively were 83% and 71%, respectively, which approximated to the percentage measured preoperatively. PAA may have the risk of rupture even though thrombosed, and preoperative measurement of the ITP might be useful to decide surgical indication. PMID- 16355098 TI - Salmonella reinfection manifesting as a fistula between the duodenum and an aortic stump pseudoaneurysm. AB - Despite the worldwide increase in Salmonella infections, a Salmonella infection of an aortic graft is an exceptional event. A Salmonella reinfection manifesting as a fistula between the duodenum and an aortic stump pseudoaneurysm 30 months after prosthetic excision, aortic ligature and extra-anatomic bypass for a Salmonella graft infection is a unique event. This unusual late complication described in this case report developed in 1 of the 5 patients whose Salmonella aortic graft infections have been previously reported. The reinfection causing septic aortitis responded to conventional surgery. PMID- 16355099 TI - Heparin-induced thrombocytopenia complicated with massive thrombosis of the inferior vena cava after filter placement. AB - A 45-year-old man presented with deep vein thrombosis of the right leg and bilateral pulmonary embolism. Heparin was administered on the initial one and a half days. On the 3rd day, an inferior vena cava (IVC) filter was placed with a heparin flush, after which massive IVC thrombosis developed. The platelet count was 221000/mm3, decreased 42% from the initial level, but remained within the normal range. Heparin was replaced by argatroban on the 13th day. The platelet count increased to 355000/mm3 on the 15th day. The patient was positive for antibody against complexes of heparin and platelet factor 4, and was diagnosed as heparin-induced thrombocytopenia with thrombosis syndrome (HITTS). When thrombosis develops during heparin treatment, it is important to suspect HITTs and to assay for the associated antibodies, regardless of the actual platelet count. PMID- 16355100 TI - Persistent sciatic artery in a patient with extracranial internal carotid artery aneurysm and infrarenal abdominal aortic aneurysm. A case report. AB - A rare case of a persistent sciatic artery (PSA) in a patient with aneurysms of the internal carotid artery and abdominal aorta is presented. A 70-year-old man was referred with intermittent claudication of the right lower extremity. Angiography and computed tomography demonstrated that this symptom was due to occlusion of the PSA. On preoperative examinations, aneurysms of the extracranial internal carotid artery and abdominal aorta were incidentally discovered, and then surgically treated prior to the management of PSA. Systemic examinations must be performed in patients with PSA in order to scrutinize associated anomalies or vascular disease. PMID- 16355101 TI - Detection of early atherosclerosis using the ultrasound parameters of the intima media thickness of the common carotid artery in families with familial combined hyperlipidaemia. PMID- 16355102 TI - Elective cesarean delivery: when is it justified? PMID- 16355103 TI - Management of neonatal abstinence syndrome in neonatal intensive care units: a national survey. AB - AIMS: To determine the monitoring and treatment of neonatal abstinence syndrome (NAS) in neonatal intensive care units (NICUs) following opiate or polydrug exposure in utero. METHODS: A pretested questionnaire was distributed via email to the chiefs of the neonatology divisions with accredited Fellowship programs in Neonatal-Perinatal Medicine in the United States. RESULTS: Of the 102 individuals contacted, 75 participated in the survey. In all, 41 of the respondents (54.5%) have a written policy regarding the management of neonatal NAS. The method of Finnegan is the most commonly used abstinence scoring system (49 of 75, 65%), while only three respondents use the Lipsitz tool. Opioids (tincture of opium, or morphine sulfate solution) are used most commonly for management of both opioid (63% of respondents) and polydrug (52% of respondents) withdrawal, followed by phenobarbital (32 % of respondents) for polydrug withdrawal and methadone (20% of respondents) for opioid withdrawal. In all, 53 respondents (70%) use phenobarbital, and 19 (25%) use intravenous morphine to control opioid withdrawal seizures, while 61 (81%) use phenobarbital in cases of polydrug withdrawal seizures. Only 53 respondents (70%) always use an abstinence scoring system to determine when to start, titrate, or terminate pharmacologic treatment of neonatal NAS. CONCLUSION: The management of neonatal psychomotor behavior consistent with withdrawal varies widely, with inconsistent policies to determine its presence or treatment. Only about half of NICUs have written guidelines for the management of NAS, which may preclude effective auditing of this practice. Educational interventions may be necessary to ensure changes in clinical practice. PMID- 16355104 TI - Fungal and bacterial sepsis and threshold ROP in preterm very low birth weight neonates. AB - OBJECTIVE: To determine whether an association exists between either fungal or bacterial sepsis and retinopathy of prematurity (ROP). STUDY DESIGN: Retrospective cohort study on all neonates with birth weight <1500 g admitted to a large Italian third Level Neonatal Intensive Care Unit in the years 1997-2001 and screened for ROP. Univariate analysis and multiple logistic regression were used to detect significant associations with ROP (all grades and threshold) in neonates with birth weight<1000 g (extremely low birth weight (ELBW)) and 1000 1500 g. RESULTS: Among 301 enrolled neonates, ROP (all grades), threshold ROP, fungal and bacterial sepsis occurred in 31.9, 12.9, 11.6 and 40.5% of the infants, respectively. At multivariate analysis, only gestational age (P=0.03), colonization by Candida non-albicans spp (P=0.03) and fungal sepsis (P=0.03) were independent predictors of threshold ROP, and only in ELBW neonates. CONCLUSIONS: Fungal (but not bacterial) sepsis is significantly and independently associated with ROP, but only in ELBW neonates and only with threshold ROP. PMID- 16355105 TI - Subcutaneous fat necrosis associated with severe hypocalcaemia in a neonate. AB - Subcutaneous fat necrosis (SFN) of the newborn is an uncommon disorder of the adipose tissue, mostly affecting full-term or post-term newborns who experience perinatal distress. The lesions of SFN typically occur during the first six weeks of life; they are usually self-limited and no specific therapy is required. The disorder may be rarely complicated with hypercalcaemia. We present the case of a neonate with perinatal asphyxia who manifested SFN followed by hypocalcaemia instead of hypercalcaemia and a biochemical profile of pseudohypoparathyroidism four weeks after the eruption of skin lesions. The infant was treated with alfacalcidiol. Blood biochemistry was normalized within one week and serum parathyroid hormone levels declined to normal over the next two months. It is suggested that perinatal asphyxia was the common etiopathogenetic factor for the development of both SFN and pseudohypoparathyroidism. PMID- 16355106 TI - Neonatal respiratory failure associated with mutation in the surfactant protein C gene. AB - We report on a term newborn with an unusual presentation and course of a rare lung disease due to mutation in SFTPC gene. This particular SFTPC mutation is novel, and the infant's lung disease was unusually severe compared to what has been previously reported in association with SFTPC mutations. PMID- 16355107 TI - Congenital epiglottic cyst presenting with severe airway obstruction at birth. PMID- 16355108 TI - Smoking and sexual dysfunction in Chinese males: findings from men's health survey. AB - To describe the prevalence of erectile dysfunction (ED) and its association with smoking among the Chinese in Hong Kong, we conducted a cross-sectional study among 819 men (aged 31-60 years) who were randomly selected among the Hong Kong residents and interviewed by trained interviewers. A structured questionnaire was used for data collection. We found that current smokers who smoked 20 cigarettes or more daily had more dissatisfaction, erection difficulty and ED than never smokers. The prevalence of dissatisfaction, difficulty in erection and ED increased significantly (P<0.05) with increasing age. Compared with never smokers, current smokers of more than 20 cigarettes daily had a greater risk of ED (age-adjusted odds ratio=1.47, 95% confidence interval: 1.00-2.16). Our results support that there are association between ED and smoking among the Chinese and suggest linking ED with smoking in the antismoking campaign and promoting smoking cessation to reduce ED among smokers. PMID- 16355109 TI - Correction of nitrergic neurovascular dysfunction in diabetic mouse corpus cavernosum by p38 mitogen-activated protein kinase inhibition. AB - Increased p38 mitogen-activated protein kinase (MAPK) in response to stress stimuli, including hyperglycemia, contributes to diabetic somatic neuropathy. However, effects on autonomic nerve and vascular function have not been determined. The aim of this study was to investigate the effects of the p38 MAPK inhibitor, LY2161793, on penile neurovascular function in streptozotocin-induced diabetic mice. Diabetes duration was 6 weeks and intervention LY2161793 treatment was given for the final 2 weeks. In vitro measurements on phenylephrine precontracted corpus cavernosum revealed a 32% reduction in maximum nitrergic nerve-mediated relaxation with diabetes that was 74% corrected by LY2161793 treatment. Maximum nitric oxide-mediated endothelium-dependent relaxation to acetylcholine was 42% attenuated by diabetes and 88% restored by LY2161793. Moreover, treatment partially corrected a diabetic deficit in endothelium independent relaxation to a nitric oxide donor. Thus, p38 MAPK inhibition corrects nitric oxide-dependent indices of diabetic erectile autonomic neuropathy and vasculopathy, a therapeutic approach potentially worthy of consideration for clinical trials. PMID- 16355110 TI - Identification of novel genes and transcription factors involved in spleen, thymus and immunological development and function. AB - We constructed and analyzed six serial analysis of gene expression (SAGE) libraries to identify genes with previously uncharacterized roles in spleen or thymus development. A total of 625 070 tags were sequenced from the three spleen (embryonic day (E)15.5, E16.5 and adult) and three thymus (E15.5, E18.5 and adult) libraries. These tags corresponded to 83 182 tag types, which mapped unambiguously to 36 133 different genes. Genes over-represented in these libraries, compared to 115 mouse SAGE libraries (www.mouseatlas.org), included genes of known and unknown immunological or developmental relevance. The expression profiles of 11 genes with unknown roles in spleen and thymus development were validated using reverse transcription-qPCR. We further characterized the expression of one of these candidates, RIKEN cDNA 9230105E10 that encodes a murine homolog of Trim5alpha, in numerous adult tissues and immune cell types. In addition, we demonstrate that transcript levels are upregulated in response to TLR stimulation of plasmacytoid dendritic cells and macrophages. This work provides the first evidence of regulated and cell type-specific expression of this gene. In addition, these observations suggest that the SAGE libraries provide an important resource for further investigations into the molecular mechanisms regulating spleen and thymus organogenesis, as well as the development of immunological competence. PMID- 16355111 TI - Variants in the gene encoding C3 are associated with asthma and related phenotypes among African Caribbean families. AB - Proinflammatory and immunoregulatory products from C3 play a major role in phagocytosis, respiratory burst, and airways inflammation. C3 is critical in adaptive immunity; studies in mice deficient in C3 demonstrate that features of asthma are significantly attenuated in the absence of C3. To test the hypothesis that the C3 gene on chromosome 19p13.3-p13.2 contains variants associated with asthma and related phenotypes, we genotyped 25 single nucleotide polymorphism (SNP) markers distributed at intervals of approximately 1.9 kb within the C3 gene in 852 African Caribbean subjects from 125 nuclear and extended pedigrees. We used the multiallelic test in the family-based association test program to examine sliding windows comprised of 2-6 SNPs. A five-SNP window between markers rs10402876 and rs366510 provided strongest evidence for linkage in the presence of linkage disequilibrium for asthma, high log[total IgE], and high log[IL 13]/[log[IFN-gamma] in terms of global P-values (P = 0.00027, 0.00013, and 0.003, respectively). A three-SNP haplotype GGC for the first three of these markers showed best overall significance for the three phenotypes (P = 0.003, 0.007, 0.018, respectively) considering haplotype-specific tests. Taken together, these results implicate the C3 gene as a priority candidate controlling risk for asthma and allergic disease in this population of African descent. PMID- 16355112 TI - Exposure to carcinogenic PAHs for the vendors of broiled food. AB - Three types of popular vendors, including meat and non-meat food (e.g, fishballs), vendor group (I), rice sausage sandwiches vendor group (II), and roasted corn on-the-cob vendor group (III), from the night markets of Taiwan were chosen for this research. The average concentrations of carcinogenic polycyclic aromatic hydrocarbons (car-PAHs) were 2445 ng/m3 for vendor (II), 2276 ng/m3 for vendor (II), and 133 ng/m3 for vendor (III). Vendors groups (I) and (II) had exposure levels of Benzo(a)pyrene (BaP)-equivalent doses (BaPeq) approximately 16 and 13 times higher than those for vendor group (III). The daily exposure doses of BaPeq in working duration for vendor group (II) and (II) were 2.80 and 2.28 microg/day, respectively. The employees of five offices on campus were chosen as a control group. The daily exposure doses of BaPeq during working hours for vendor group (II) and (II) were about 22 times and 18 times, respectively, higher than those for control group. This exposure dose for vendors group (III) (0.18 microg/day) was comparable with the dose for control group (0.13 microg/day). The whole-day exposure doses of t-PAHs, car-PAHs, BaP, and BaPeq for vendor groups (I) and (II) were very close to those during daily working hours. However, the daily exposure doses of BaPeq during working hours for vendor group (III) occurred only in a proportion of 62% of the whole-day exposure doses. PMID- 16355113 TI - Allele-specific RNA interference for neurological disease. AB - Suppressing the expression of toxic genes through RNAi holds great promise for the treatment of human disease. Allele-specific approaches have now been used to silence dominant toxic genes implicated in several neurological disorders. Here, we review strategies used to achieve allele-specific silencing in light of recent developments in the field of RNAi biology. In particular, new insights into siRNA and miRNA processing may be used to improve efficiency and specificity of RNAi therapy. We further discuss steps that can be taken to maximize the therapeutic benefits of this powerful technology. PMID- 16355114 TI - Woodchuck hepatitis virus post-transcriptional regulatory element deleted from X protein and promoter sequences enhances retroviral vector titer and expression. AB - Introduction of the post-transcriptional regulatory element (PRE) of woodchuck hepatitis virus (WHV) into the 3' untranslated region of retroviral and lentiviral gene transfer vectors enhances both titer and transgene expression. Optimal use of the PRE is often necessary to obtain vectors with sufficient performance for therapeutic applications. The enhancing activity of the PRE depends on the precise configuration of its sequence and the context of the vector and cell into which it is introduced. However, data obtained in the context of WHV-associated hepatocellular carcinomas suggests that the PRE might potentially contribute to tumorigenesis, especially if encoding a truncated version of the WHV X protein. Oncogenic side effects of lentiviral vectors containing the PRE have reinforced these safety concerns, although a causal role of the PRE remained unproven. Here, we demonstrate that PRE mutants can be generated that are devoid of X protein open reading frames (ORFs) as well as other ORFs exceeding 25 amino acids, without significant loss of RNA enhancement activity. Furthermore, the X protein promoter could be deleted without compromising the enhancement of vector titers and transgene expression. Such a modified PRE sequence appears useful for future vector design. PMID- 16355115 TI - Induction of effective therapeutic antitumor immunity by direct in vivo administration of lentiviral vectors. AB - Ex vivo lentivirally transduced dendritic cells (DC) have been described to induce CD8+ and CD4+ T-cell responses against various tumor-associated antigens (TAAs) in vitro and in vivo. We report here that direct administration of ovalbumin (OVA) encoding lentiviral vectors caused in vivo transduction of cells that were found in draining lymph nodes (LNs) and induced potent anti-OVA cytotoxic T cells similar to those elicited by ex vivo transduced DC. The cytotoxic T-lymphocyte (CTL) response following direct injection of lentiviral vectors was highly effective in eliminating target cells in vivo up to 30 days after immunization and was efficiently recalled after a boost immunization. Injection of lentiviral vectors furthermore activated OVA-specific CD4+ T cells and this CD4 help was shown to be necessary for an adequate primary and memory CTL response. When tested in therapeutic tumor experiments with OVA+ melanoma cells, direct administration of lentiviral vectors slowed down tumor growth to a comparable extent with the highest dose of ex vivo transduced DC. Taken together, these data indicate that direct in vivo administration of lentiviral vectors encoding TAAs has strong potential for anticancer vaccination. PMID- 16355116 TI - Human neural progenitors deliver glial cell line-derived neurotrophic factor to parkinsonian rodents and aged primates. AB - Glial cell line-derived neurotrophic factor (GDNF) has been shown to increase the survival and functioning of dopamine neurons in a variety of animal models and some recent human trials. However, delivery of any protein to the brain remains a challenge due to the blood/brain barrier. Here we show that human neural progenitor cells (hNPC) can be genetically modified to release glycosylated GDNF in vitro under an inducible promoter system. hNPC-GDNF were transplanted into the striatum of rats 10 days following a partial lesion of the dopamine system. At 2 weeks following transplantation, the cells had migrated within the striatum and were releasing physiologically relevant levels of GDNF. This was sufficient to increase host dopamine neuron survival and fiber outgrowth. At 5 weeks following grafting there was a strong trend towards functional improvement in transplanted animals and at 8 weeks the cells had migrated to fill most of the striatum and continued to release GDNF with transport to the substantia nigra. These cells could also survive and release GDNF 3 months following transplantation into the aged monkey brain. No tumors were found in any animal. hNPC can be genetically modified, and thereby represent a safe and powerful option for delivering growth factors to specific targets within the central nervous system for diseases such as Parkinson's. PMID- 16355117 TI - Chick embryo lethal orphan virus can be polymer-coated and retargeted to infect mammalian cells. AB - Non-human adenovirus vectors have attractive immunological properties for gene therapy but are frequently restricted by inefficient transduction of human target cells. Using chicken embryo lethal orphan (CELO) virus, we employed a nongenetic mechanism of polymer coating and retargeting with basic fibroblast growth factor (bFGF-pc-CELOluc), a strategy that permits efficient tropism modification of human adenovirus. bFGF-pc-CELOluc showed efficient uptake and transgene expression in chick embryo fibroblasts (CEF), and increased levels of binding and internalization in a variety of human cell lines. Transgene expression was also greater than unmodified CELOluc in PC-3 human prostate cells, although the specific activity (RLU per internalized viral genome) was decreased. In CEF, the specific activity of bFGF-pc-CELOluc was considerably higher than in the human prostate cell line PC-3. Retargeted virus was fully resistant to inhibition by human serum with known adenovirus-neutralizing activity in vitro, while in mice CELOluc was cleared less rapidly from the blood than Adluc following i.v. administration in the presence of adenovirus neutralizing serum. Polymer coating and retargeting with bFGF further reduced rates of clearance for both viruses, suggesting protection against both neutralizing and opsonizing factors. The data indicate that CELO virus may be retargeted to infect human cells via alternative, potentially disease-specific, receptors and resist the effects of pre-existing humoral immunity. PMID- 16355118 TI - Awareness of the Japanese Society of Hypertension Guidelines for the Management of Hypertension (JSH 2000) and compliance to its recommendations: surveys in 2000 and 2004. AB - Clinic physicians' awareness of the Japanese hypertension guideline (JSH 2000) and compliance with its recommendations were assessed to derive policy implications for effective blood pressure control. Data were obtained from two postal questionnaire surveys conducted in 2000 and 2004, and subjects were 896 and 1425 clinic physicians, respectively, who were engaged in general internal medicine. Recognition rates of JSH 2000 were 63.1% (n = 822) before its announcement in 2000 and 94.4% (n = 1400) in 2004. Rates of access, familiarity and utilisation of JSH 2000 were 87.0, 81.6 and 68.9%, respectively (n = 1400) in 2004. As for major management strategies for low-risk hypertension: in 2000, for patients with 140-149/90-94 mmHg, 81.5% of 812 respondents performed lifestyle modification, and 11.2% prescribed medicines, whereas for patients with 150 159/95-99 mmHg, 71.7% of 807 respondents prescribed medicines, and 24.3% conducted lifestyle modification; in 2004, 90.0% of 1384 respondents conducted lifestyle modification, 22.6% prescribed medicines, 2.5% referred patients to other facilities, and 6.4% did nothing. In 2004, 68.9% of 1388 respondents agreed with the new definition of hypertension, whereas 17.1% preferred 160/95 mmHg. Respondents' age (P<0.05) and a percentage of hypertensives in daily patient load (P<0.0005) significantly associated with the choice of the old criteria. In conclusion, JSH 2000 achieved a substantial improvement in clinic physicians' awareness and their compliance to its recommendations on low-risk hypertension management. One of the strategies for further enhancement in their compliance with JSH 2004 would be its dissemination to those who are old and/or do not see hypertensive patients so frequently. PMID- 16355119 TI - Use of home blood pressure measurement by hypertensive patients in Southern Greece. PMID- 16355120 TI - Influence of target organ lesion detection (assessment of microalbuminuria and echocardiogram) in cardiovascular risk stratification and treatment of untreated hypertensive patients. AB - European guidelines indicate the importance of the evaluation of global cardiovascular risk (CVR) to determine the management of the hypertensive patients (EH). However, in primary care, the diagnostic work-up (PCD) only includes the metabolic risk factors. The aim of this study was to assess the importance of microalbuminuria (MA) and echocardiogram (ECHO) in the process of risk stratification, and the number of patients to be treated with drugs at diagnosis. In total, 155 nontreated EH were included in the study. Blood pressure, a lipid profile and plasma glucose (LG) were determined after an overnight fast. MA was evaluated with dipstick MICRALTEST, and in those patients with two positive results, it was measured again in two 24-h urine samples and was considered positive (MA+) if the average was >30 mg/24 h. Left ventricular mass index was calculated and values>125 g/m2 were considered as LV hypertrophy (LVH+). When the patients were stratified according to PCD, 22 had to be treated with drugs. When MA, ECHO and both tests used together were added to the risk evaluation, the number of patients to be treated were 42, 51 and 64, respectively (P<0.001 vs PCD). It is mainly in patients who have moderate cardiovascular risk that risk changes, whereas risk hardly changes in those having low and very high risk. In conclusion, in EH with moderate risk, measurement of MA, due to its easy availability and low cost, seems to be a cost effective screening test to avoid the underestimation of the CVR. PMID- 16355121 TI - Interaction between pulse wave velocity, augmentation index, pulse pressure and left ventricular function in chronic heart failure. AB - Pulse wave velocity (PWV), the carotid augmentation index (AIx), and pulse pressure (PP) may be prognostic factors in heart failure, but the possible influence of the ejection fraction (EF) and other simple haemodynamic variables on them has not been investigated in this setting. Noninvasive methods were used to measure carotid-radial (CR), carotid-femoral (CF) PWV and AIx, and brachial PP, in 135 consecutive patients with stable symptomatic chronic heart failure. The patients were divided into two groups, with preserved (>or=40%) or reduced (<40%) EF. CF-PWV, AIx and PP were lower in the decreased EF group (8.85+/-2.77 versus 10.60+/-2.75 m/s, P<0.001; 121+/-21 versus 132+/-24, P=0.009 and 41+/-19 versus 67+/-17 mmHg, P<0.001), but CR-PWV values were similar regardless of the EF status. These results were not modified after adjustment for age and sex. Multiple regression analysis showed that AIx and PP were systematically related to time domain parameters (heart rate or ejection duration) and EF, whatever the group. CF-PWV was weakly related to time domain values and unrelated to mean blood pressure (BP) or EF in the preserved EF group, whereas it was related to both mean BP and EF in the low EF group. In conclusion, whatever the EF level, PP and AIx were strongly modulated in the time domain, by pressure and by the EF level. The same relationships were found with CF-PWV, but only in the reduced EF group. Whether CF-PWV is the best prognostic factor in patients with 'diastolic' heart failure must be confirmed in a prospective study. PMID- 16355136 TI - Top-down versus step-up approaches to chronic inflammatory bowel disease: presumed innocent or presumed guilty. PMID- 16355137 TI - Role of genetic screening in identifying susceptibility to acute fatty liver of pregnancy. PMID- 16355138 TI - Selecting livers for transplantation: can beggars be choosers? PMID- 16355151 TI - Is esomeprazole useful for the treatment of upper gastrointestinal symptoms in patients on NSAIDs? PMID- 16355152 TI - How useful are proton-pump inhibitors for diagnosis and therapy of patients with noncardiac chest pain? PMID- 16355153 TI - How effective is Enteryx implantation for the treatment of patients with GERD? PMID- 16355155 TI - Is ursodeoxycholic acid effective for the prevention of colorectal adenoma recurrence? PMID- 16355154 TI - Incidence of adverse reactions to azathioprine in patients with Crohn's disease. PMID- 16355156 TI - How safe and effective is endoscopic full-thickness plication for the treatment of patients with GERD? PMID- 16355157 TI - The etiology, assessment, and treatment of fecal incontinence. AB - Fecal incontinence is a common symptom that often impairs quality of life. It is generally caused by a variety of conditions that are associated with anorectal sensorimotor dysfunction and/or diarrhea. Assessment should be tailored to age and symptom severity. Modulation of disordered bowel habits is the key to management; biofeedback and surgery might also be beneficial. PMID- 16355158 TI - Endoscopic band ligation in the treatment of portal hypertension. AB - The evidence that endoscopic band ligation (EBL) has greater efficacy and fewer side effects than endoscopic injection sclerotherapy has renewed interest in endoscopic treatments for portal hypertension. The introduction of multishot band devices, which allow the placement of 5-10 bands at a time, has made the technique much easier to perform, avoiding the use of overtubes and their related complications. EBL sessions are usually repeated at 2 week intervals until varices are obliterated, which is achieved in about 90% of patients after 2-4 sessions. Variceal recurrence is frequent, with 20-75% of patients requiring repeated EBL sessions. According to current evidence, nonselective beta-blockers are the preferred treatment option for prevention of a first variceal bleed, whereas EBL should be reserved for patients with contraindications or intolerance to beta-blockers. Nonselective beta-blockers, probably in association with the vasodilator isosorbide mononitrate, and EBL are good treatment options to prevent recurrent variceal rebleeding. The efficacy of EBL might be increased by combining it with beta-blocker therapy. Patients who are intolerant, have contraindications or bled while receiving primary prophylaxis with beta-blockers must be treated with EBL. In the latter situation, EBL should be added to rather than replace beta-blocker therapy. EBL, in combination with vasoactive drugs, is the recommended form of therapy for acute esophageal variceal bleeding; however, endoscopic injection sclerotherapy can be used in the acute setting if EBL is technically difficult. PMID- 16355159 TI - Mechanisms of disease: nicotine--a review of its actions in the context of gastrointestinal disease. AB - Smoking tobacco is associated with a number of gastrointestinal disorders. In some, such as Crohn's disease and peptic ulcer disease, it increases the risk of disease and has a detrimental effect on their course. In others, such as ulcerative colitis, it decreases the risk of disease and can have a favorable effect on disease course and severity. In the eighteenth and nineteenth centuries, nicotine was used as a 'panacea' for various ailments, including abdominal symptoms--it is now under investigation to elucidate its role in gastrointestinal diseases that are associated with smoking. The actions of nicotine are complex; it is likely that its effects on the central nervous system, gastrointestinal tract and immune system interact with other risk factors, such as genetic susceptibility, to influence disease outcomes. This review focuses on the mechanisms of action of nicotine that might be relevant in gastrointestinal disease. PMID- 16355160 TI - Acute deterioration of a woman following acute pancreatitis with pseudocysts. AB - BACKGROUND: A 49-year-old white female with a history of acute pancreatitis that was complicated by pseudocysts presented with severe acute-onset abdominal pain of a few hours duration with associated vomiting. Physical examination showed a soft abdomen with mild diffuse tenderness and positive bowel sounds. Initial blood work revealed a drop in her hematocrit, and elevated but stable amylase and lipase levels. A CT scan of the abdomen revealed a splenic artery pseudoaneurysm with extravasation of contrast medium into an adjacent pseudocyst. INVESTIGATIONS: Physical examination, blood analysis and a CT scan of the abdomen with contrast medium. DIAGNOSIS: Splenic artery pseudoaneurysm bleeding into an adjacent pseudocyst. MANAGEMENT: Embolization of the splenic artery across the neck of the pseudoaneurysm. PMID- 16355161 TI - Retinal angiogenesis in development and disease. AB - The retina has long been regarded as 'an approachable part of the brain' for investigating neurosensory processes. Cell biologists are now capitalizing on the accessibility of the retina to investigate important aspects of developmental angiogenesis, including how it relates to neuronal and glial development, morphogenesis, oxygen sensing and progenitor cells. Pathological angiogenesis also occurs in the retina and is a major feature of leading blinding diseases, particularly diabetic retinopathy. The retina and its clinical disorders have a pivotal role in angiogenesis research and provide model systems in which to investigate neurovascular relationships and angiogenic diseases. PMID- 16355163 TI - Seismology: tectonic strain in plate interiors? AB - It is not fully understood how or why the inner areas of tectonic plates deform, leading to large, although infrequent, earthquakes. Smalley et al. offer a potential breakthrough by suggesting that surface deformation in the central United States accumulates at rates comparable to those across plate boundaries. However, we find no statistically significant deformation in three independent analyses of the data set used by Smalley et al., and conclude therefore that only the upper bounds of magnitude and repeat time for large earthquakes can be inferred at present. PMID- 16355167 TI - Europe's right stuff. PMID- 16355168 TI - Preparing for disaster. PMID- 16355169 TI - Wiki's wild world. PMID- 16355173 TI - Fake pottery buries theory of early start for Christianity. PMID- 16355172 TI - Big money for cancer genomics. PMID- 16355174 TI - Developing nations offer hope in climate talks. PMID- 16355175 TI - Budget boost gets Russia back in the space game. PMID- 16355177 TI - Pokemon blocks gene name. PMID- 16355178 TI - Journal grows suspicious of Vioxx data. PMID- 16355179 TI - Snapshot: blot on the landscape. PMID- 16355180 TI - Internet encyclopaedias go head to head. PMID- 16355183 TI - Natural disasters: the chaos to come. PMID- 16355184 TI - Natural disasters: the vanishing coast. PMID- 16355185 TI - Natural disasters: roots of recovery. PMID- 16355187 TI - Olympus finds market rival hard to swallow. PMID- 16355189 TI - Languages: land may speak louder than words. PMID- 16355190 TI - Languages: Europe puts its money where its mouth is. PMID- 16355191 TI - Climate research opponent is not a friend to science. PMID- 16355197 TI - An asymmetric world. PMID- 16355192 TI - Biodiversity: there's a role to be played by 'museum-keepers' too. PMID- 16355199 TI - Fluid dynamics: drat such custard! PMID- 16355198 TI - Archaeology: life on the Costa del Cromer. PMID- 16355200 TI - Neuroscience: a painful factor. PMID- 16355202 TI - Earth science: volte-face in the Punjab. PMID- 16355203 TI - Developmental biology: a message to the back side. PMID- 16355204 TI - Microbiology: perspectives on plague. PMID- 16355205 TI - Obituary: Richard Southwood (1931-2005). PMID- 16355206 TI - Post-spawning egg care by a squid. AB - Gonatus onyx is one of the most abundant cephalopods in the Pacific and Atlantic Oceans and is an important prey species for a variety of vertebrate predators, but a full understanding of its life history has been hampered because spawning occurs at great depths, where observation is difficult. Here we describe post spawning egg care, or brooding, in this deep-sea squid. Our finding is unexpected because this behaviour differs from the reproductive habits of all other known squid species. PMID- 16355208 TI - Colloid science: non-spherical bubbles. AB - Surface tension gives gas bubbles their perfect spherical shape by minimizing the surface area for a given volume. Here we show that gas bubbles and liquid drops can exist in stable, non-spherical shapes if the surface is covered, or 'armoured', with a close-packed monolayer of particles. When two spherical armoured bubbles are fused, jamming of the particles on the interface supports the unequal stresses that are necessary to stabilize a non-spherical shape. PMID- 16355210 TI - Angiogenesis in life, disease and medicine. AB - The growth of blood vessels (a process known as angiogenesis) is essential for organ growth and repair. An imbalance in this process contributes to numerous malignant, inflammatory, ischaemic, infectious and immune disorders. Recently, the first anti-angiogenic agents have been approved for the treatment of cancer and blindness. Angiogenesis research will probably change the face of medicine in the next decades, with more than 500 million people worldwide predicted to benefit from pro- or anti-angiogenesis treatments. PMID- 16355211 TI - Endothelial cells and VEGF in vascular development. AB - The intricate patterning processes that establish the complex vascular system during development depend on a combination of intrinsic pre-patterning and extrinsic responses to environmental parameters. Mutational studies in mice and fish have shown that the vascular system is highly sensitive to genetic disruption and have identified potential targets for therapeutic interventions. New insights into non-vascular roles of vascular endothelial growth factor and the requirement for endothelial cells in adult organs and stem-cell niches highlight possible side effects of anti-angiogenic therapy and the need for new targets. PMID- 16355212 TI - Lymphangiogenesis in development and human disease. AB - The lymphatic vasculature forms a vessel network that drains interstitial fluid from tissues and returns it to the blood. Lymphatic vessels are also an essential part of the body's immune defence. They have an important role in the pathogenesis of several diseases, such as cancer, lymphoedema and various inflammatory conditions. Recent biological and technological developments in lymphatic vascular biology will lead to a better understanding and treatment of these diseases. PMID- 16355213 TI - From angiogenesis to neuropathology. AB - Angiogenesis--the growth of new blood vessels--is a crucial force for shaping the nervous system and protecting it from disease. Recent advances have improved our understanding of how the brain and other tissues grow new blood vessels under normal and pathological conditions. Angiogenesis factors, especially vascular endothelial growth factor, are now known to have roles in the birth of new neurons (neurogenesis), the prevention or mitigation of neuronal injury (neuroprotection), and the pathogenesis of stroke, Alzheimer's disease and motor neuron disease. As our understanding of pathophysiology grows, these developments may point the way towards new molecular and cell-based therapies. PMID- 16355214 TI - Angiogenesis as a therapeutic target. AB - Inhibiting angiogenesis is a promising strategy for treatment of cancer and several other disorders, including age-related macular degeneration. Major progress towards a treatment has been achieved over the past few years, and the first antiangiogenic agents have been recently approved for use in several countries. Therapeutic angiogenesis (promoting new vessel growth to treat ischaemic disorders) is an exciting frontier of cardiovascular medicine, but further understanding of the mechanisms of vascular morphogenesis is needed first. PMID- 16355215 TI - Probing ion-channel pores one proton at a time. AB - Although membrane proteins often rely on ionizable residues for structure and function, their ionization states under physiological conditions largely elude experimental estimation. To gain insight into the effect of the local microenvironment on the proton affinity of ionizable residues, we have engineered individual lysines, histidines and arginines along the alpha-helical lining of the transmembrane pore of the nicotinic acetylcholine receptor. We can detect individual proton binding-unbinding reactions electrophysiologically at the level of a single proton on a single side chain as brief blocking-unblocking events of the passing cation current. Kinetic analysis of these fluctuations yields the position-dependent rates of proton transfer, from which the corresponding pK(a) values and shifts in pK(a) can be calculated. Here we present a self-consistent, residue-by-residue description of the microenvironment around the pore-lining transmembrane alpha-helices (M2) in the open-channel conformation, in terms of the excess free energy that is required to keep the engineered basic side chains protonated relative to bulk water. A comparison with closed-channel data leads us to propose that the rotation of M2, which is frequently invoked as a hallmark of the gating mechanism of Cys-loop receptors, is minimal, if any. PMID- 16355216 TI - Hypomethylation-linked activation of PAX2 mediates tamoxifen-stimulated endometrial carcinogenesis. AB - Tamoxifen, a selective oestrogen receptor modulator, has been used in the treatment of all stages of hormone-responsive breast cancer. However, tamoxifen shows partial oestrogenic activity in the uterus and its use has been associated with an increased incidence of endometrial cancer. The molecular explanation for these observations is not known. Here we show that tamoxifen and oestrogen have distinct but overlapping target gene profiles. Among the overlapping target genes, we identify a paired-box gene, PAX2, that is crucially involved in cell proliferation and carcinogenesis in the endometrium. Our experiments show that PAX2 is activated by oestrogen and tamoxifen in endometrial carcinomas but not in normal endometrium, and that this activation is associated with cancer-linked hypomethylation of the PAX2 promoter. PMID- 16355217 TI - The afterglow and elliptical host galaxy of the short gamma-ray burst GRB 050724. AB - Despite a rich phenomenology, gamma-ray bursts (GRBs) are divided into two classes based on their duration and spectral hardness--the long-soft and the short-hard bursts. The discovery of afterglow emission from long GRBs was a watershed event, pinpointing their origin to star-forming galaxies, and hence the death of massive stars, and indicating an energy release of about 10(51) erg. While theoretical arguments suggest that short GRBs are produced in the coalescence of binary compact objects (neutron stars or black holes), the progenitors, energetics and environments of these events remain elusive despite recent localizations. Here we report the discovery of the first radio afterglow from the short burst GRB 050724, which unambiguously associates it with an elliptical galaxy at a redshift z = 0.257. We show that the burst is powered by the same relativistic fireball mechanism as long GRBs, with the ejecta possibly collimated in jets, but that the total energy release is 10-1,000 times smaller. More importantly, the nature of the host galaxy demonstrates that short GRBs arise from an old (> 1 Gyr) stellar population, strengthening earlier suggestions and providing support for coalescing compact object binaries as the progenitors. PMID- 16355218 TI - An origin in the local Universe for some short gamma-ray bursts. AB - Gamma-ray bursts (GRBs) divide into two classes: 'long', which typically have initial durations of T90 > 2 s, and 'short', with durations of T90 < 2 s (where T90 is the time to detect 90% of the observed fluence). Long bursts, which on average have softer gamma-ray spectra, are known to be associated with stellar core-collapse events-in some cases simultaneously producing powerful type Ic supernovae. In contrast, the origin of short bursts has remained mysterious until recently. A subsecond intense 'spike' of gamma-rays during a giant flare from the Galactic soft gamma-ray repeater, SGR 1806-20, reopened an old debate over whether some short GRBs could be similar events seen in galaxies out to approximately 70 Mpc (refs 6-10; redshift z approximately 0.016). Shortly after that, localizations of a few short GRBs (with optical afterglows detected in two cases) have shown an apparent association with a variety of host galaxies at moderate redshifts. Here we report a correlation between the locations of previously observed short bursts and the positions of galaxies in the local Universe, indicating that between 10 and 25 per cent of short GRBs originate at low redshifts (z < 0.025). PMID- 16355219 TI - An origin for short gamma-ray bursts unassociated with current star formation. AB - Two short (< 2 s) gamma-ray bursts (GRBs) have recently been localized and fading afterglow counterparts detected. The combination of these two results left unclear the nature of the host galaxies of the bursts, because one was a star forming dwarf, while the other was probably an elliptical galaxy. Here we report the X-ray localization of a short burst (GRB 050724) with unusual gamma-ray and X ray properties. The X-ray afterglow lies off the centre of an elliptical galaxy at a redshift of z = 0.258 (ref. 5), coincident with the position determined by ground-based optical and radio observations. The low level of star formation typical for elliptical galaxies makes it unlikely that the burst originated in a supernova explosion. A supernova origin was also ruled out for GRB 050709 (refs 3, 31), even though that burst took place in a galaxy with current star formation. The isotropic energy for the short bursts is 2-3 orders of magnitude lower than that for the long bursts. Our results therefore suggest that an alternative source of bursts--the coalescence of binary systems of neutron stars or a neutron star-black hole pair--are the progenitors of short bursts. PMID- 16355220 TI - Chaos and threshold for irreversibility in sheared suspensions. AB - Systems governed by time reversible equations of motion often give rise to irreversible behaviour. The transition from reversible to irreversible behaviour is fundamental to statistical physics, but has not been observed experimentally in many-body systems. The flow of a newtonian fluid at low Reynolds number can be reversible: for example, if the fluid between concentric cylinders is sheared by boundary motion that is subsequently reversed, then all fluid elements return to their starting positions. Similarly, slowly sheared suspensions of solid particles, which occur widely in nature and science, are governed by time reversible equations of motion. Here we report an experiment showing precisely how time reversibility fails for slowly sheared suspensions. We find that there is a concentration dependent threshold for the deformation or strain beyond which particles do not return to their starting configurations after one or more cycles. Instead, their displacements follow the statistics of an anisotropic random walk. By comparing the experimental results with numerical simulations, we demonstrate that the threshold strain is associated with a pronounced growth in the Lyapunov exponent (a measure of the strength of chaotic particle interactions). The comparison illuminates the connections between chaos, reversibility and predictability. PMID- 16355221 TI - Reorganization of the western Himalayan river system after five million years ago. AB - Uplift of mountains driven by tectonic forces can influence regional climate as well as regional drainage patterns, which in turn control the discharge of eroded sediment to the ocean. But the nature of the interactions between tectonic forces, climate and drainage evolution remains contested. Here we reconstruct the erosional discharge from the Indus river over the past 30 million years using seismic reflection data obtained from drill core samples from the Arabian Sea and neodymium isotope data. We find that the source of the Indus sediments was dominated by erosion within and north of the Indus suture zone until five million years ago; after that, the river began to receive more erosional products from Himalayan sources. We propose that this change in the erosional pattern is caused by a rerouting of the major rivers of the Punjab into the Indus, which flowed east into the Ganges river before that time. Seismic reflection profiles from the Indus fan suggest high mass accumulation rates during the Pleistocene epoch partly driven by increased drainage to the Indus river after five million years ago and partly by faster erosion linked to a stronger monsoon over the past four million years. Our isotope stratigraphy for the Indus fan provides strong evidence for a significant change in the geometry of western Himalayan river systems in the recent geologic past. PMID- 16355222 TI - Efficacy of the post-perovskite phase as an explanation for lowermost-mantle seismic properties. AB - Constraining the chemical, rheological and electromagnetic properties of the lowermost mantle (D'') is important to understand the formation and dynamics of the Earth's mantle and core. To explain the origin of the variety of characteristics of this layer observed with seismology, a number of theories have been proposed, including core-mantle interaction, the presence of remnants of subducted material and that D'' is the site of a mineral phase transformation. This final possibility has been rejuvenated by recent evidence for a phase change in MgSiO3 perovskite (thought to be the most prevalent phase in the lower mantle) at near core-mantle boundary temperature and pressure conditions. Here we explore the efficacy of this 'post-perovskite' phase to explain the seismic properties of the lowermost mantle through coupled ab initio and seismic modelling of perovskite and post-perovskite polymorphs of MgSiO3, performed at lowermost mantle temperatures and pressures. We show that a post-perovskite model can explain the topography and location of the D'' discontinuity, apparent differences in compressional- and shear-wave models and the observation of a deeper, weaker discontinuity. Furthermore, our calculations show that the regional variations in lower-mantle shear-wave anisotropy are consistent with the proposed phase change in MgSiO3 perovskite. PMID- 16355223 TI - The earliest record of human activity in northern Europe. AB - The colonization of Eurasia by early humans is a key event after their spread out of Africa, but the nature, timing and ecological context of the earliest human occupation of northwest Europe is uncertain and has been the subject of intense debate. The southern Caucasus was occupied about 1.8 million years (Myr) ago, whereas human remains from Atapuerca-TD6, Spain (more than 780 kyr ago) and Ceprano, Italy (about 800 kyr ago) show that early Homo had dispersed to the Mediterranean hinterland before the Brunhes-Matuyama magnetic polarity reversal (780 kyr ago). Until now, the earliest uncontested artefacts from northern Europe were much younger, suggesting that humans were unable to colonize northern latitudes until about 500 kyr ago. Here we report flint artefacts from the Cromer Forest-bed Formation at Pakefield (52 degrees N), Suffolk, UK, from an interglacial sequence yielding a diverse range of plant and animal fossils. Event and lithostratigraphy, palaeomagnetism, amino acid geochronology and biostratigraphy indicate that the artefacts date to the early part of the Brunhes Chron (about 700 kyr ago) and thus represent the earliest unequivocal evidence for human presence north of the Alps. PMID- 16355224 TI - A RhoGDP dissociation inhibitor spatially regulates growth in root hair cells. AB - Root hairs are cellular protuberances extending from the root surface into the soil; there they provide access to immobile inorganic ions such as phosphate, which are essential for growth. Their cylindrical shape results from a polarized mechanism of cell expansion called tip growth in which elongation is restricted to a small area at the surface of the hair-forming cell (trichoblast) tip. Here we identify proteins that spatially control the sites at which cell growth occurs by isolating Arabidopsis mutants (scn1) that develop ectopic sites of growth on trichoblasts. We cloned SCN1 and showed that SCN1 is a RhoGTPase GDP dissociation inhibitor (RhoGDI) that spatially restricts the sites of growth to a single point on the trichoblast. We showed previously that localized production of reactive oxygen species by RHD2/AtrbohC NADPH oxidase is required for hair growth; here we show that SCN1/AtrhoGDI1 is a component of the mechanism that focuses RHD2/AtrbohC-catalysed production of reactive oxygen species to hair tips during wild-type development. We propose that the spatial organization of growth in plant cells requires the local RhoGDI-regulated activation of the RHD2/AtrbohC NADPH oxidase. PMID- 16355225 TI - BDNF from microglia causes the shift in neuronal anion gradient underlying neuropathic pain. AB - Neuropathic pain that occurs after peripheral nerve injury depends on the hyperexcitability of neurons in the dorsal horn of the spinal cord. Spinal microglia stimulated by ATP contribute to tactile allodynia, a highly debilitating symptom of pain induced by nerve injury. Signalling between microglia and neurons is therefore an essential link in neuropathic pain transmission, but how this signalling occurs is unknown. Here we show that ATP stimulated microglia cause a depolarizing shift in the anion reversal potential (E(anion)) in spinal lamina I neurons. This shift inverts the polarity of currents activated by GABA (gamma-amino butyric acid), as has been shown to occur after peripheral nerve injury. Applying brain-derived neurotrophic factor (BDNF) mimics the alteration in E(anion). Blocking signalling between BDNF and the receptor TrkB reverses the allodynia and the E(anion) shift that follows both nerve injury and administration of ATP-stimulated microglia. ATP stimulation evokes the release of BDNF from microglia. Preventing BDNF release from microglia by pretreating them with interfering RNA directed against BDNF before ATP stimulation also inhibits the effects of these cells on the withdrawal threshold and E(anion). Our results show that ATP-stimulated microglia signal to lamina I neurons, causing a collapse of their transmembrane anion gradient, and that BDNF is a crucial signalling molecule between microglia and neurons. Blocking this microglia-neuron signalling pathway may represent a therapeutic strategy for treating neuropathic pain. PMID- 16355226 TI - Heat activation of TRPM5 underlies thermal sensitivity of sweet taste. AB - TRPM5, a cation channel of the TRP superfamily, is highly expressed in taste buds of the tongue, where it has a key role in the perception of sweet, umami and bitter tastes. Activation of TRPM5 occurs downstream of the activation of G protein-coupled taste receptors and is proposed to generate a depolarizing potential in the taste receptor cells. Factors that modulate TRPM5 activity are therefore expected to influence taste. Here we show that TRPM5 is a highly temperature-sensitive, heat-activated channel: inward TRPM5 currents increase steeply at temperatures between 15 and 35 degrees C. TRPM4, a close homologue of TRPM5, shows similar temperature sensitivity. Heat activation is due to a temperature-dependent shift of the activation curve, in analogy to other thermosensitive TRP channels. Moreover, we show that increasing temperature between 15 and 35 degrees C markedly enhances the gustatory nerve response to sweet compounds in wild-type but not in Trpm5 knockout mice. The strong temperature sensitivity of TRPM5 may underlie known effects of temperature on perceived taste in humans, including enhanced sweetness perception at high temperatures and 'thermal taste', the phenomenon whereby heating or cooling of the tongue evoke sensations of taste in the absence of tastants. PMID- 16355227 TI - Mapping stem cell activities in the feather follicle. AB - It is important to know how different organs 'manage' their stem cells. Both hair and feather follicles show robust regenerative powers that episodically renew the epithelial organ. However, the evolution of feathers (from reptiles to birds) and hairs (from reptiles to mammals) are independent events and their follicular structures result from convergent evolution. Because feathers do not have the anatomical equivalent of a hair follicle bulge, we are interested in determining where their stem cells are localized. By applying long-term label retention, transplantation and DiI tracing to map stem cell activities, here we show that feather follicles contain slow-cycling long-term label-retaining cells (LRCs), transient amplifying cells and differentiating keratinocytes. Each population, located in anatomically distinct regions, undergoes dynamic homeostasis during the feather cycle. In the growing follicle, LRCs are enriched in a 'collar bulge' niche. In the moulting follicle, LRCs shift to populate a papillar ectoderm niche near the dermal papilla. On transplantation, LRCs show multipotentiality. In a three-dimensional view, LRCs are configured as a ring that is horizontally placed in radially symmetric feathers but tilted in bilaterally symmetric feathers. The changing topology of stem cell activities may contribute to the construction of complex feather forms. PMID- 16355228 TI - The zebrafish dorsal axis is apparent at the four-cell stage. AB - A central question in the development of multicellular organisms pertains to the timing and mechanisms of specification of the embryonic axes. In many organisms, specification of the dorsoventral axis requires signalling by proteins of the Transforming growth factor-beta and Wnt families. Here we show that maternal transcripts of the zebrafish Nodal-related morphogen, Squint (Sqt), can localize to two blastomeres at the four-cell stage and predict the dorsal axis. Removal of cells containing sqt transcripts from four-to-eight-cell embryos or injection of antisense morpholino oligonucleotides targeting sqt into oocytes can cause a loss of dorsal structures. Localization of sqt transcripts is independent of maternal Wnt pathway function and requires a highly conserved sequence in the 3' untranslated region. Thus, the dorsoventral axis is apparent by early cleavage stages and may require the maternally encoded morphogen Sqt and its associated factors. Because the 3' untranslated region of the human nodal gene can also localize exogenous sequences to dorsal cells, this mechanism may be evolutionarily conserved. PMID- 16355229 TI - The Rae1-Nup98 complex prevents aneuploidy by inhibiting securin degradation. AB - Cdc20 and Cdh1 are the activating subunits of the anaphase-promoting complex (APC), an E3 ubiquitin ligase that drives cells into anaphase by inducing degradation of cyclin B and the anaphase inhibitor securin. To prevent chromosome missegregation, APC activity directed against these mitotic regulators must be inhibited until all chromosomes are properly attached to the mitotic spindle. Here we show that in mitosis timely destruction of securin by APC is regulated by the nucleocytoplasmic transport factors Rae1 and Nup98. We show that combined Rae1 and Nup98 haploinsufficiency in mice results in premature separation of sister chromatids, severe aneuploidy and untimely degradation of securin. We find that Rae1 and Nup98 form a complex with Cdh1-activated APC (APC(Cdh1)) in early mitosis and specifically inhibit APC(Cdh1)-mediated ubiquitination of securin. Dissociation of Rae1 and Nup98 from APC(Cdh1) coincides with the release of the mitotic checkpoint protein BubR1 from Cdc20-activated APC (APC(Cdc20)) at the metaphase to anaphase transition. Together, our results suggest that Rae1 and Nup98 are temporal regulators of APC(Cdh1) that maintain euploidy by preventing unscheduled degradation of securin. PMID- 16355232 TI - Let's get critical. PMID- 16355230 TI - Towards complete cofactor arrangement in the 3.0 A resolution structure of photosystem II. AB - Oxygenic photosynthesis in plants, algae and cyanobacteria is initiated at photosystem II, a homodimeric multisubunit protein-cofactor complex embedded in the thylakoid membrane. Photosystem II captures sunlight and powers the unique photo-induced oxidation of water to atmospheric oxygen. Crystallographic investigations of cyanobacterial photosystem II have provided several medium resolution structures (3.8 to 3.2 A) that explain the general arrangement of the protein matrix and cofactors, but do not give a full picture of the complex. Here we describe the most complete cyanobacterial photosystem II structure obtained so far, showing locations of and interactions between 20 protein subunits and 77 cofactors per monomer. Assignment of 11 beta-carotenes yields insights into electron and energy transfer and photo-protection mechanisms in the reaction centre and antenna subunits. The high number of 14 integrally bound lipids reflects the structural and functional importance of these molecules for flexibility within and assembly of photosystem II. A lipophilic pathway is proposed for the diffusion of secondary plastoquinone that transfers redox equivalents from photosystem II to the photosynthetic chain. The structure provides information about the Mn4Ca cluster, where oxidation of water takes place. Our study uncovers near-atomic details necessary to understand the processes that convert light to chemical energy. PMID- 16355233 TI - Evidence-based dentistry in German-language dental journals 1997-2003. AB - The purpose of this systematic literature search was to evaluate how the evidence based approach appears in German-language dental journals. All volumes of 28 dental journals published between 1997 and 2003 were searched by hand; for 15 of these journals, an additional electronic search was made. The focus of the search was all articles, editorials or letters to the Editor dealing with topics related to evidence-based dentistry (EBD)/medicine or using its terminology. Of the 1776 journal issues that were scrutinised, 368 relevant publications were found. After a low, albeit slightly increasing, number of pertinent contributions published between 1997 and 1999, there was a sharp rise in 2000 and 2001, culminating in 2003. Our findings indicate a rising awareness of the concept of EBD in the German-language dental literature. Nonetheless, its reception within the German speaking dental community is still modest. PMID- 16355234 TI - Fluoridated milk may be beneficial to schoolchildren by helping prevent caries. PMID- 16355235 TI - Flowable restorative system has a higher retention rate than conventional resin sealant on primary teeth. PMID- 16355236 TI - Periodontal disease, preterm birth and low birthweight. PMID- 16355237 TI - Rapid maxillary expansion treatment could produce long-term transverse skeletal changes. PMID- 16355238 TI - Rapid maxillary expansion treatment could produce long-term dental arch changes. PMID- 16355239 TI - Third molars may have a negative impact on periodontal health. PMID- 16355240 TI - Ten-year survival rate for cantilevered fixed partial dentures. AB - DATA SOURCES: PubMed (1966-April 2004) provided the primary data source along with the bibliographies from identified articles and reviews. A manual search of two relevant journals (Clinical Implant Dentistry and Related Research, Clinical Oral Implants Research, International Journal of Oral & Maxillofacial Implants, International Journal of Periodontics and Restorative Dentistry, International Journal of Prosthodontics, Journal of Clinical Periodontology, Journal of Periodontology and Journal of Prosthetic Dentistry published; published 2001 2003) provided a further source of data. STUDY SELECTION: Because there were no randomised controlled trials (RCT) that compared fixed partial dentures (FPD) with and without cantilever extensions, other studies were selected if they met the following criteria: were published in the English language; were prospective and retrospective cohort studies; had a mean follow-up of >5 years; included patients who were clinically examined at follow-up; and reported details on suprastructures and described at least one-third of reconstructions as FPD. DATA EXTRACTION AND SYNTHESIS: Two independent reviewers screened articles for inclusion. Disagreements were resolved by discussion and agreement determined by kappa. Two reviewers extracted data on the survival and success of the reconstructions and on biological and technical complications. Failure and complication rates were calculated by dividing the number of events by the total exposure time. RESULTS: Three prospective and 10 retrospective cohort studies incorporating 700 patients and 816 FPD were included. The mean number of cantilever extensions ranged from 1.1 to 6.0. Meta-analyses gave an estimated survival, after 10 years, of 81.8% [95% confidence interval (CI), 78.2-84.9] and a proportion success of 63% (95% CI, 54.7-70.2). Considering biological complications using Poisson model analyses, the estimated rate of FPD lost because of caries after 10 years was 3.1% (95%CI, 1.0-8.8) and that for loss of vitality was 32.6% (95% CI, 13.9-64.9). The rate of FPD lost because of periodontitis was 1% (95% CI, 0.3-3.0). Cumulative 10-year complication rates were: 2.9% (95% CI, 1.7-5.0) for fracture of abutment tooth; 2.4% (95% CI, 0.6 9.8) for rate of loss as a result of abutment fracture; 16.1% (95% CI, 8.8-28.4) for loss of retention; and 5.9% (95% CI, 3.3-10.4) for material complications. CONCLUSIONS: Success and survival rates for cantilever FPD are poorer than those for conventional FPD and this is accompanied by frequent biological and technical complications. PMID- 16355241 TI - Ninety-four per cent of combined tooth-implant fixed partial dentures survive 5 years. PMID- 16355242 TI - Implant survival in augmented maxillary sinus is more variable than that of implants placed in posterior maxilla. PMID- 16355243 TI - No "gold standard" critical appraisal tool for allied health research. PMID- 16355244 TI - Study design II. Issues of chance, bias, confounding and contamination. AB - In the first article in the series I explained the importance of study design and gave an overview of the main types of design. Here, I describe the ways in which the results of a study may deviate from the truth and the measures that can be taken to help minimise this when designing a study. PMID- 16355250 TI - Cure therapeutics and strategic prevention: raising the bar for mental health research. AB - Mental disorders cause more disability than any other class of medical illness in Americans between ages 15 and 44 years. The suicide rate is higher than the annual mortality from homicide, AIDS, and most forms of cancer. In contrast to nearly all communicable and most non-communicable diseases, there is little evidence that the morbidity and mortality from mental disorders have changed in the past several decades. Mental health advocates, including psychiatric researchers, have pointed to stigma as one of the reasons for the lack of progress with mental illnesses relative to other medical illnesses. This review considers how the expectations and goals of the research community have contributed to this relative lack of progress. In contrast to researchers in cancer and heart disease who have sought cures and preventions, biological psychiatrists in both academia and industry have set their sights on incremental and marketable advances, such as drugs with fewer adverse effects. This essay argues for approaches that can lead to cures and strategies for prevention of schizophrenia and mood disorders. PMID- 16355251 TI - Biological, cellular, and molecular characteristics of an inducible transgenic skin tumor model: a review. AB - The genetically initiated Tg.AC transgenic mouse carries a transgene consisting of an oncogenic v-Ha-ras coding region flanked 5' by a mouse zeta-globin promoter and 3' by an SV-40 polyadenylation sequence. Located on chromosome 11, the transgene is transcriptionally silent until activated by chemical carcinogens, UV light, or full-thickness wounding. Expression of the transgene is an early event that drives cellular proliferation resulting in clonal expansion and tumor formation, the unique characteristics now associated with the Tg.AC mouse. This ras-dependent phenotype has resulted in the widespread interest and use of the Tg.AC mouse in experimental skin carcinogenesis and as an alternative carcinogenesis assay. This review examines the general biology of the tumorigenic responses observed in Tg.AC mice, the genetic interactions of the ras transgene, and explores the cellular and molecular regulation of zeta-globin promoted transgene expression. As a prototype alternative model to the current long-term rodent bioassays, the Tg.AC has generated a healthy discussion on the future of transgenic bioassays, and opened the doors for subsequent models for toxicity testing. The further exploration and elucidation of the molecular controls of transgene expression will enhance the usefulness of this mouse and enable a better understanding of the Tg.AC's discriminate response to chemical carcinogens. PMID- 16355252 TI - Clines, clusters, and the effect of study design on the inference of human population structure. AB - Previously, we observed that without using prior information about individual sampling locations, a clustering algorithm applied to multilocus genotypes from worldwide human populations produced genetic clusters largely coincident with major geographic regions. It has been argued, however, that the degree of clustering is diminished by use of samples with greater uniformity in geographic distribution, and that the clusters we identified were a consequence of uneven sampling along genetic clines. Expanding our earlier dataset from 377 to 993 markers, we systematically examine the influence of several study design variables--sample size, number of loci, number of clusters, assumptions about correlations in allele frequencies across populations, and the geographic dispersion of the sample--on the "clusteredness" of individuals. With all other variables held constant, geographic dispersion is seen to have comparatively little effect on the degree of clustering. Examination of the relationship between genetic and geographic distance supports a view in which the clusters arise not as an artifact of the sampling scheme, but from small discontinuous jumps in genetic distance for most population pairs on opposite sides of geographic barriers, in comparison with genetic distance for pairs on the same side. Thus, analysis of the 993-locus dataset corroborates our earlier results: if enough markers are used with a sufficiently large worldwide sample, individuals can be partitioned into genetic clusters that match major geographic subdivisions of the globe, with some individuals from intermediate geographic locations having mixed membership in the clusters that correspond to neighboring regions. PMID- 16355253 TI - Revealing posttranscriptional regulatory elements through network-level conservation. AB - We used network-level conservation between pairs of fly (Drosophila melanogaster/D. pseudoobscura) and worm (Caenorhabditis elegans/C. briggsae) genomes to detect highly conserved mRNA motifs in 3' untranslated regions. Many of these elements are complementary to the 5' extremity of known microRNAs (miRNAs), and likely correspond to their target sites. We also identify known targets of RNA-binding proteins, and many novel sites not yet known to be functional. Coherent sets of genes with similar function often bear the same conserved elements, providing new insights into their cellular functions. We also show that target sites for distinct miRNAs are often simultaneously conserved, suggesting combinatorial regulation by multiple miRNAs. A genome-wide search for conserved stem-loops, containing complementary sequences to the novel sites, revealed many new candidate miRNAs that likely target them. We also provide evidence that posttranscriptional networks have undergone extensive rewiring across distant phyla, despite strong conservation of regulatory elements themselves. PMID- 16355255 TI - [Pediatricians and the external causes of morbidity and mortality]. PMID- 16355254 TI - Folding free energies of 5'-UTRs impact post-transcriptional regulation on a genomic scale in yeast. AB - Using high-throughput technologies, abundances and other features of genes and proteins have been measured on a genome-wide scale in Saccharomyces cerevisiae. In contrast, secondary structure in 5'-untranslated regions (UTRs) of mRNA has only been investigated for a limited number of genes. Here, the aim is to study genome-wide regulatory effects of mRNA 5'-UTR folding free energies. We performed computations of secondary structures in 5'-UTRs and their folding free energies for all verified genes in S. cerevisiae. We found significant correlations between folding free energies of 5'-UTRs and various transcript features measured in genome-wide studies of yeast. In particular, mRNAs with weakly folded 5'-UTRs have higher translation rates, higher abundances of the corresponding proteins, longer half-lives, and higher numbers of transcripts, and are upregulated after heat shock. Furthermore, 5'-UTRs have significantly higher folding free energies than other genomic regions and randomized sequences. We also found a positive correlation between transcript half-life and ribosome occupancy that is more pronounced for short-lived transcripts, which supports a picture of competition between translation and degradation. Among the genes with strongly folded 5' UTRs, there is a huge overrepresentation of uncharacterized open reading frames. Based on our analysis, we conclude that (i) there is a widespread bias for 5' UTRs to be weakly folded, (ii) folding free energies of 5'-UTRs are correlated with mRNA translation and turnover on a genomic scale, and (iii) transcripts with strongly folded 5'-UTRs are often rare and hard to find experimentally. PMID- 16355256 TI - [Injury control from the perspective of contextual pediatrics]. AB - OBJECTIVE: To describe the relationship between injury control and contextual pediatrics. SOURCES OF DATA: Quasi-systematic review of MEDLINE, SciELO and LILACS databases, using combinations of the words contextual, community, injury, accident and violence; and non-systematic review of book chapters and classic articles. SUMMARY OF THE FINDINGS: Safety depends on the interaction of family habits, cultural patterns and surroundings. Contextual pediatrics sees the child, the family, and the community as a continuum; health diagnosis (sequential observation of problems and assets) is one of its cornerstones. Changing intrapersonal factors for injuries requires the use of both passive and active strategies. Family and cultural risk factors for injury: home overcrowding, moving, poverty, and young, illiterate and unemployed parents. The main neighborhood factors: material deprivation and traffic. Cultural factors: illiteracy, unsafe products, lack of mass transportation, handguns, workplaces without safety rules, faulty community organization, lack of communication between social sectors, inadequate legislation, low priority for safety among government actions, lack of economic resources, and low academic commitment with the field of safety. CONCLUSIONS: The pediatrician's roles include strengthening of the longitudinal relationship with families, integrated interdisciplinary work, constructive intervention, partnership with community, counseling on injury risks pertaining to each developmental stage, by using lists with explicit processes and contents, and by handing out written materials. Active advocacy for safety promotion in different environments, besides the clinical setting. PMID- 16355257 TI - [Safe community]. AB - OBJECTIVE: To analyze the effectiveness of prevention programs for intentional and unintentional injuries, based on safe community principles, in children and adolescents. SOURCES OF DATA: An electronic search was performed in the MEDLINE and LILACS database, corresponding to the period from 2000 to 2005. For prevention of unintentional injuries, we used the following keywords: injury and prevention and community or population and intervention. For prevention of intentional injuries, we used the following keywords: violence and prevention and community or population and intervention; the age range from zero to 18 years was used as a threshold. The inclusion criteria were: original articles which evaluated the effectiveness of interventions using more than one strategy, involving a whole community or group, published in Portuguese, English or Spanish. SUMMARY OF THE FINDINGS: 152 studies were obtained as a result. Considering the inclusion criteria, five articles were found on unintentional injury prevention, one about intentional injury prevention and one paper addressing these two issues. All studies analyzed demonstrate benefits to children and adolescents, but in different proportions. CONCLUSIONS: A restricted number of programs using the safe community concept were found. The investigations analyzed in this study presented positive results. Increased efforts to further develop this evidence are still needed, respecting local characteristics, and developing evaluation indicators that allow for a better comparison between different studies. PMID- 16355258 TI - [Unintentional injuries in the home environment: home safety]. AB - OBJECTIVE: To review the characteristics of unintentional injuries and their impact on children and adolescents. SOURCES OF DATA: Articles published between 2000 and 2005 in the MEDLINE, EBSCO, Proquest, SciELO, BVS and Google Scholar databases were selected. The authors used the keywords unintentional injuries, injuries, safe home, burns, falls, drowning, scorpions, snakes, poisoning, child, adolescent, mortality, injury control, and hospitalization. Some articles were evaluated based on the selected publications. SUMMARY OF THE FINDINGS: Unintentional injuries in the world and in Brazil are analyzed, and so are the behaviors currently adopted for injury prevention and control. The impact on mortality, on physical damage, and the economic burden of injuries are evaluated. Special emphasis is placed on home environment, approaching the effects of child development, social disparities and contextualization of home environment on children's world and vulnerabilities. The main types of events that cause physical damage to the child and adolescent in the home environment are described. CONCLUSION: The prevention of injuries in the home environment is possible. In this case, health professionals have the challenge to reduce the consequences of unintentional injuries on the morbidity and mortality of children and young people in Brazil and in the whole world. PMID- 16355259 TI - [Safe school]. AB - OBJECTIVE: To review the strategies to make school a safe environment. The paper first addresses the social context of accidents and violence in the school environment, and makes recommendations, based on the literature data, for the implementation of safe schools. SOURCE OF DATA: Articles published between 1993 and 2005 in the MEDLINE database. Brazilian epidemiological and literature data have also been searched. SUMMARY OF THE FINDINGS: There is growing evidence that intervention has multiple components, focusing on health education practices, with the participation of the whole community. The aim of those interventions is to help students and community members to adopt healthy and safe behaviors. Schools are taking on an increasing role in health promotion, disease prevention, and injury prevention. In the context of prevention of external causes of morbidity and mortality, it is important to recognize a risky environment, places, and risk behaviors as favorable to injury and violence, as well as the concept of accident as something one can avoid. CONCLUSION: Implementation of safe schools represents a promising new direction for school-based preventive work. It is important to note that a safe school should intervene not only in its physical structure, but it should also make it as safe as possible by gathering the school community through health education, and mainly encouraging healthy behavior. PMID- 16355260 TI - [Bullying--aggressive behavior among students]. AB - OBJECTIVE: To warn pediatricians about the high prevalence of bullying among students, to raise their awareness about the importance of their action in the prevention, diagnosis, and treatment of possible damage to children s health and development, and about the necessity to instruct families and society on how to face the most frequent form of youth violence. SOURCE OF DATA: Bibliographic databases and relevant Internet sites were searched for recent articles and texts about the theme. SUMMARY OF THE FINDINGS: Aggressive behavior among students is a universal problem, traditionally accepted as natural and usually disregarded or not given proper attention by adults. Studies carried out during the past two decades showed that bullying can have immediate and late negative outcomes for children and adolescents who are directly or indirectly involved. The adoption of continued preventive programs in grade schools and in junior high schools has demonstrated to be one of the most effective measures for the prevention of alcohol and drug consumption and for the reduction of social violence. CONCLUSION: The prevention of bullying among students represents an essential public health measure that may allow for total children s development, qualifying them for a healthy and safe social coexistence. PMID- 16355261 TI - [Follow-up of child abuse victims: challenges for the pediatrician]. AB - OBJECTIVE: To review practical questions about the initial assistance and follow up of child abuse victims and their families by pediatricians. SOURCES OF DATA: A literature review was carried out using the MEDLINE and LILACS databases, including the years 2000 to 2005. Some articles from past years and books were included due to their importance. SUMMARY OF THE FINDINGS: Initial assistance is one of the most important actions by health professionals for the protection of abused children in different healthcare sectors (community, outpatient clinics, emergency rooms and infirmary), and it is fundamental for the reduction of immediate and long-term negative consequences of violence. The protection services cannot monitor all the families under their responsibility and most child abuse cases are not even reported to those institutions; therefore, regular follow-up by a pediatrician is advisable. It is important to provide the family with support and guidance until the child is safe. The main challenges are: to be involved without causing more violence; to consider all the family as the focus of attention, including the family members who have committed the assault, helping them to change inadequate behaviors; to develop specific abilities to carry out this work, which must be multiprofessional, interdisciplinary and intersectoral. CONCLUSIONS: Families face difficulties when their children are abused and when the situation gains notoriety, demanding interventions from many institutions. In this process, a pediatrician can guide and help them to guarantee the protection and healthy development of their children. To overcome challenges, health professionals have to be technically and emotionally prepared. PMID- 16355262 TI - [The pediatrician and traffic safety]. AB - OBJECTIVE: To instruct children and teenagers about safe transportation and about their behavior as cyclists and pedestrians. SOURCES OF DATA: The MEDLINE and LILACS databases were searched, using the following terms, from 1995 to 2005: traffic accidents, child, adolescent, morbidity, mortality, age, and gender. Scientific articles on morbidity and mortality from traffic accidents, children in a pedestrian situation, as cyclists or as motor vehicle passengers, were selected. SUMMARY OF THE FINDINGS: More than 300 scientific articles were found, but only the studies that addressed those issues in a broad manner were selected. CONCLUSIONS: Traffic-related mortality rates are still very high and show the necessity to intensify educational campaigns for community awareness, in addition to encouraging investments in effective pedestrian safety. The pediatrician s role is to decrease the number of children and adolescents who get involved in traffic accidents by providing them with traffic guidance and education. PMID- 16355263 TI - [Urban violence: a challenge for pediatricians]. AB - OBJECTIVES: To present the main aspects of an approach to urban violence among children and adolescents and to point out the social and educational role of pediatricians. SOURCES OF DATA: A literature review based on MEDLINE, LILACS and SciELO was carried out for the years 1993 to 2005, using the following keywords: urban violence, children and adolescents. In addition to the review, policies and institutional reports on violence were also analyzed. SUMMARY OF THE FINDINGS: The causal relationship of violence is presented in a range of different ways, from personal points of view to broader structural aspects. The literature suggests that urban violence results from varied actions, and also from specific risk behaviors. It is a worrying and complex phenomenon that results in high levels of morbidity and mortality, affecting children and adolescents. Special attention was given to homicide and to the psychological results of violent acts. Firearms are the most lethal instruments among adolescents and young males when compared to all other causes of death. CONCLUSIONS: Urban violence is one of the main social problems in Brazil. Violence prevention requires intersectoral and multiprofessional actions with the participation of the government and of the organized civil society. Children, and above all adolescents, are the groups that are most widely exposed to the consequences of urban violence. Pediatricians can have an important role in the process of prevention, diagnosis and treatment. Through their wide-ranging abilities, pediatricians are well-positioned to help victims and their families to establish a healthy and dynamic relationship with their environment and with themselves. PMID- 16355264 TI - [Current view of sexual abuse in childhood and adolescence]. AB - OBJECTIVE: To review single aspects, which involve sexual abuse in childhood and adolescence, giving subsidies for accurate diagnosis and management, emphasizing the short and long-term consequences. SOURCES OF DATA: National and international literature review of the MEDLINE and LILACS databases, using abuse and sexual violence as keywords (1988 to 2005), in addition to the authors theoretical and practical experiences. SUMMARY OF THE FINDINGS: Sexual abuse has a great impact on child and adolescent physical and mental health, affecting development and causing lifelong damage. Its early detection allows for adequate treatment and follow-up, with minimum consequences. Family involvement must be taken into consideration, and any kind of improper relationship should be evaluated and treated, so that it can be discouraged and eliminated; otherwise, intergenerational abuse will result and probably recur. The identification of domestic violence and of physical and psychological warning signs concerning sexual abuse is part of the general evaluation. CONCLUSIONS: Pediatricians should be able to identify the signs and symptoms sexually abused children and adolescents carry with them. They have to listen carefully to the patients, perform the necessary clinical examination and establish the proper treatment. They should also be able to deal with psychological aspects and have enough knowledge about the laws and about legal and social protection measures, which are always very useful in assisting victims of sexual violence. PMID- 16355265 TI - [Environmental chemical hazards and child health]. AB - OBJECTIVES: To review the recent medical literature on environmental chemical hazards to child health. SOURCES OF DATA: Articles published on this subject between 1999 and 2005 were searched in the MEDLINE database. Books, manuals and statements on child environmental health, issued by institutions such as the American Academy of Pediatrics and the World Health Organization, were also reviewed. SUMMARY OF THE FINDINGS: There has been a growing concern in the last few years with the exposure of children to environmental chemicals. Around 85,000 synthetic chemicals are produced today, and 2,800 of them are mass-produced. There is little knowledge regarding their effects on developing organisms. Children have a greater exposure to environmental pollutants than adults, because their metabolic needs and behaviors (e.g.: crawling, bringing objects to the mouth, playing closer to the ground) put them at special risk of contact with chemicals when they breathe, eat, drink or play. Heavy metals, pesticides, persistent organic pollutants and, at home, environmental tobacco smoke have been associated with the increasing number of diseases such as asthma, neurodevelopmental disorders and childhood cancer. CONCLUSION: Screening of risk situations using tools such as Environmental History has been stimulated alongside a greater commitment of pediatricians towards measures that can reduce the exposure of children and adolescents to environmental chemicals. PMID- 16355266 TI - [Acute toxic exposure in children: an overview]. AB - OBJECTIVE: To review the literature on acute toxic exposure in children, excluding envenomations. SOURCES OF DATA: MEDLINE review (emphasis on the past decade), including the American Academy of Clinical Toxicology and the European Association of Poison Centres and Clinical Toxicologists position statements and position papers (peer-reviewed information based on scientific evidence and broad consensus) on gastrointestinal decontamination, multiple-dose activated charcoal and urine alkalinization. SUMMARY OF THE FINDINGS: Acute toxic exposure in children is a common event, mainly in children under six years of age. Death is rare. Although widely employed, there is no evidence that gastrointestinal decontamination and multiple-dose activated charcoal improve the outcome of poisoned patients. Very few efficient antidotes are used on a consistent basis, and some of them are very expensive and not available in Brazil. CONCLUSIONS: Ipecac syrup and cathartics should not be administered on a routine basis in acute toxic exposures in outpatient treatment. Excluding the contraindications, single-dose activated charcoal and gastric lavage may be considered within one hour of ingestion if a patient ingested a potentially toxic amount or a potentially lethal amount, respectively. Whole bowel irrigation, multiple-dose activated charcoal and urine alkalinization may be considered in a few situations. Fomepizole and octreotide are safe and efficient antidotes, which can be used in the treatment of alcohol (methanol and ethylene glycol) and sulfonylureas poisoning, respectively. PMID- 16355267 TI - [Initial assessment and transportation of an injured child]. AB - OBJECTIVES: Since trauma has a great impact on mortality and preventable morbidity among children and adolescents, in addition to its social consequences, the aim of this study is to evaluate peculiarities regarding prehospital, in hospital, and interhospital assessment of injured pediatric patients. SOURCES OF DATA: The Cochrane database was searched for systematic reviews and controlled trials, the MEDLINE and LILACS databases were used for the last 5 years, and review of older reference sections in significant publications using trauma, pediatric trauma, primary survey, secondary survey, in-hospital assessment, prehospital and transportation as keywords. SUMMARY OF THE FINDINGS: There are different strategies that constitute the systematic assessment of the injured pediatric patient. Nevertheless, almost all strategies involve knowing the peculiarities about the pediatric population, with regard to age, to the growth and development process, and finally, to the minimally acceptable and available resources. CONCLUSIONS: The most important principle is to do no further harm. If the peculiarities about airway maintenance, ventilation, circulation with hemorrhage control, disability, exposure up to secondary survey and preparation of prehospital and interhospital transportation are properly taken care of, better results will certainly be obtained. PMID- 16355270 TI - Four and half lim protein 2 (FHL2) stimulates osteoblast differentiation. AB - FHL2, a molecule that interacts with many integrins and transcription factors, was found to play an important role in osteoblast differentiation. Overexpression of FHL2 increases the accumulation of osteoblast differentiation markers and matrix mineralization, whereas FHL2 deficiency results in inhibition of osteoblast differentiation and decreased bone formation. INTRODUCTION: Integrin matrix interaction plays a critical role in osteoblast function. It has been shown that the cytoplasmic domains of integrin beta subunits mediate signal transduction induced by integrin-matrix interaction. We reasoned that the identification of proteins interacting with beta-cytoplasmic tails followed by analysis of the function of these proteins would enhance our understanding on integrin signaling and the roles of these proteins in osteoblast activities. MATERIALS AND METHODS: Yeast two hybrid assay was used to identify proteins interacting with the cytoplasmic domain of integrin beta5 subunit. The association of these proteins with integrin alphavbeta5 was confirmed by confocal analysis and co-immunoprecipitation. A stable MC3T3-E1 cells line overexpressing Four and Half Lim Protein 2 (FHL2) and mouse osteoblasts deficient in FHL2 were used to study the roles of FHL2 in osteoblast differentiation and bone formation. Matrix protein expression was determined by mRNA analysis and Western blotting. Matrix mineralization was detected by Alizarin red staining. Alkaline phosphatase activity was also measured. muCT was used to determine bone histomorphometry. RESULTS AND CONCLUSIONS: FHL2 and actin-binding proteins, palladin and filamin A, were identified as proteins interacting with beta5 cytoplasmic domain. FHL2 co localized with alphavbeta5 at the focal adhesion sites in association with palladin and filamin A. FHL2 was also present in nuclei. Osteoblasts overexpressing FHL2 exhibited increased adhesion to and migration on matrix proteins. Conversely, FHL2 stimulation of CREB activity was dependent on integrin function because it was inhibited by Gly-Arg-Gly-Asp-Ser (GRGDS) peptide. The expression of osteoblast differentiation markers and Msx2 was upregulated, and bone matrix mineralization was increased in FHL2 overexpressing cells. In contrast, FHL2-deficient bone marrow cells and osteoblasts displayed decreased osteoblast colony formation and differentiation, respectively, compared with wildtype cells. Moreover, FHL2-deficient female mice exhibited greater bone loss than the wildtype littermates after ovariectomy. Thus, FHL2 plays an important role in osteoblast differentiation and bone formation. PMID- 16355269 TI - Smad3-deficient chondrocytes have enhanced BMP signaling and accelerated differentiation. AB - Smad3 deficiency accelerates chondrocyte maturation and leads to osteoarthritis. Primary chondrocytes without Smad3 lack compensatory increases of TGF-beta signaling factors, but BMP-related gene expression is increased. Smad2 or Smad3 overexpression and BMP blockade abrogate accelerated maturation in Smad3-/- chondrocytes. BMP signaling is increased in TGF-beta deficiency and is required for accelerated chondrocyte maturation. INTRODUCTION: Disruption of TGF-beta signaling results in accelerated chondrocyte maturation and leads to postnatal dwarfism and premature osteoarthritis. The mechanisms involved in this process were studied using in vitro murine chondrocyte cultures. MATERIALS AND METHODS: Primary chondrocytes were isolated from the sterna of neonatal wildtype and Smad3 /- mice. Expressions of maturational markers, as well as genes involved in TGF beta and BMP signaling were examined. Chondrocytes were treated with TGF-beta and BMP-2, and effects on maturation-related genes and BMP/TGF-beta responsive reporters were examined. Recombinant noggin or retroviral vectors expressing Smad2 or Smad3 were added to the cultures. RESULTS: Expression of colX and other maturational markers was markedly increased in Smad3-/- chondrocytes. Smad3-/- chondrocytes lacked compensatory increases in Smad2, Smad4, TGFRII, Sno, or Smurf2 and had reduced expression of TGF-beta1 and TGFRI. In contrast, Smad1, Smad5, BMP2, and BMP6 expression was increased, suggesting a shift from TGF-beta toward BMP signaling. In Smad3-/- chondrocytes, alternative TGF-beta signaling pathways remained responsive, as shown by luciferase assays. These non-Smad3 dependent TGF-beta pathways reduced colX expression and alkaline phosphatase activity in TGF-beta-treated Smad3-/- cultures, but only partially. In contrast, Smad3-/- chondrocytes were more responsive to BMP-2 treatment and had increased colX expression, phosphoSmads 1, 5, and 8 levels, and luciferase reporter activity. Overexpression of both Smad2 and Smad3 blocked spontaneous maturation in Smad3-deficient chondrocytes. Maturation was also abrogated by the addition of noggin, an extracellular BMP inhibitor. CONCLUSIONS: These findings show a key role for BMP signaling during the chondrocyte maturation, occurring with loss of TGF-beta signaling with important implications for osteoarthritis and cartilage diseases. PMID- 16355271 TI - Immunocompetent properties of human osteoblasts: interactions with T lymphocytes. AB - We sought to determine whether osteoblasts (OBs) can serve as accessory cells (ACs) for T-cell activation and whether T cells directly activate OB production of IL-6, using primary human OBs (NHOst), the transformed fetal osteoblast line hFOB1.19, and an osteosarcoma line SaOS-2. Robust, bidirectional activating interactions were shown using each of these three human ostoblast lines. INTRODUCTION: Osteoblasts (OBs) could come into contact with lymphocytes during inflammatory joint destruction and fracture repair. MATERIALS AND METHODS: We used several in vitro assays to assess the ability of T cells and OBs to interact in the generation of immune and inflammatory responses. RESULTS: By flow cytometry, three OB cell lines all were found to express ligands for T-cell co stimulation. The integrin ligand CD54/ICAM-1 was constitutively expressed by hFOB1.19 and NHOst and was upregulated on SaOS-2 by IFN-gamma. MHC Class II was upregulated on all three lines by IFN-gamma. CD166/ALCAM, a ligand of the T-cell molecule CD6, was constitutively expressed on all three lines. A second putative CD6 ligand designated 3A11 was expressed on hFOB1.19 and NHOst, but not consistently on SaOS-2. The ectoenzyme CD26 (dipeptidyl peptidase IV) was expressed on hFOB1.19 and NHOst, but not on SaOS-2. All three cell lines presented superantigen to T cells, especially after treatment with IFN-gamma. Superantigen presentation was inhibited by antibodies to the leukocyte integrin CD11a/CD18 (LFA-1), MHC Class II, and CD54/ICAM-1. T cells, particularly when cytokine activated for 7 days before co-culture, stimulated all three osteoblast lines to produce interleukin (IL)-6, and this effect was boosted when IL-17 was added to the co-cultures with either resting T cells or cytokine-activated T cells. CONCLUSIONS: Bidirectional activating interactions are readily shown between human T cells and several types of human OBs. The expression by OBs of ligands for the T cell-specific molecule CD6, as well as other molecules involved in immune interactions, strongly suggests that such in vitro interactions are representative of physiologic or pathologic events that occur in vivo. PMID- 16355272 TI - Immune regulation of 25-hydroxyvitamin-D3-1alpha-hydroxylase in human monocytes. AB - Monocytes express 1alpha-hydroxylase, the enzyme responsible for final hydroxylation of vitamin D3, in response to IFNgamma and CD14/TLR4 activation. Cross-talk between the JAK-STAT, the NF-kappaB, and the p38 MAPK pathways is necessary, and direct binding of C/EBPbeta to its recognition sites in the promoter of the 1alpha-hydroxylase gene is a prerequisite. INTRODUCTION: The activated form of vitamin D3, 1,25(OH)2D3, known for its action in bone and mineral homeostasis, has important immunomodulatory effects. 1,25(OH)2D3 modulates the immune system through specific nuclear receptors, whereas macrophages produce 1,25(OH)2D3. In monocytes, the expression of 1alpha hydroxylase, the enzyme responsible for final hydroxylation of vitamin D3, is regulated by immune stimuli. The aim of this study was to elucidate the intracellular pathways through which interferon (IFN)gamma and Toll-like receptor (TLR) modulation regulate expression of 1alpha-hydroxylase in monocytes/macrophages. MATERIALS AND METHODS: Monocytes were isolated from peripheral blood mononuclear cells (PBMCs) and stimulated with IFNgamma (12.5 U/ml) and/or lipopolysaccharide (LPS; 100 ng/ml) for 48 h. The following inhibitors were used: janus kinase (JAK) inhibitor AG490 (50 microM), NF-kappaB inhibitor sulfasalazine (0.25 mM), p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 (5 microM). 1alpha-hydroxylase mRNA expression was monitored by qRT-PCR. Phosphorylation of transcription factors was studied by Western blotting. Transfection of mutated or deletion promoter constructs, cloned in the pGL3-luciferase reporter plasmid, were performed in the RAW264.7 cell line. Cells were stimulated with IFNgamma (100 U/ml) and LPS (100 microg/ml), and promoter activity was studied. Binding of signal transducer and activator of transcription (STAT)1alpha, NF-kappaB, and C/EBPbeta to their respective binding sites in the promoter was analyzed by gel shift assays. RESULTS: 1alpha-hydroxylase mRNA expression in monocytes is synergistically induced by IFNgamma and CD14/TLR4 ligation and paralleled by 1,25(OH)2D3 production. This induction requires the JAK-STAT, NF-kappaB, and p38 MAPK pathways. Each of them is essential, because blocking individual pathways is sufficient to block 1alpha-hydroxylase expression (JAK inhibitor, 60% inhibition, p < 0.01; NF-kappaB inhibitor, 70% inhibition, p < 0.05; p38 MAPK inhibitor, 95% inhibition, p < 0.005). In addition, we show the involvement of the p38 MAPK pathway in phosphorylation of C/EBPbeta. Direct binding of C/EBPbeta to its recognition sites in the 1alpha-hydroxylase promoter is necessary to enable its immune-stimulated upregulation. CONCLUSION: IFNgamma and CD14/TLR4 binding regulate expression of 1alpha-hydroxylase in monocytes in a synergistic way. Combined activation of the JAK-STAT, p38 MAPK, and NF-kappaB pathways is necessary, with C/EBPbeta most probably being the essential transcription factor controlling immune-mediated transcription. PMID- 16355273 TI - CCR1 acts downstream of NFAT2 in osteoclastogenesis and enhances cell migration. AB - We found that a chemokine receptor gene, CCR1, acts downstream of NFAT2 in RANKL stimulated RAW264 and bone marrow cells. The upstream regulatory region of CCR1 showed RANKL-dependent and CsA-suppressible promoter activity. Downregulation of the expression and function of CCR1 suppressed cell migration. INTRODUCTION: We previously reported that the expression of NFAT2 induced by RANKL is a key process for progression to multinucleated cells in an in vitro osteoclastogenesis system. Identifying the target genes of NFAT2 would thus be informative about the differentiation process. We focused here on chemokine and chemokine receptor genes that act downstream of NFAT2 in RAW264 cells as well as osteoclast precursors prepared from bone marrow cells. MATERIALS AND METHODS: RAW264 mouse monocyte/macrophage line cells were cultured with or without cyclosporin A (CsA) in the presence of RANKL or glutathione S-transferase (GST). Osteoclast precursors were prepared from bone marrow cells. RANKL-inducible and CsA suppressible genes were searched for by microarray analysis, and expression was confirmed by quantitative RT-PCR. Promoter activity was measured by luciferase gene reporter assay. Short interfering (si)RNA for CCR1 was introduced in RAW264 cells. Cell migration activity was examined using a Boyden chamber assay. RESULTS AND CONCLUSIONS: We identified the chemokine receptor gene CCR1 as a gene showing significant differential expression profiles in osteoclastogenesis in the presence versus the absence of CsA, an inhibitor of NFAT. This property was unique to CCR1 among the chemokine and chemokine receptor genes examined in both RAW264 and bone marrow cells. The upstream regulatory region was isolated from CCR1, and its RANKL-dependent and CsA-suppressible promoter activity was confirmed. The functional significance of CCR1 was assessed by monitoring the migration of cells in a transwell migration assay, and this activity was abolished when either CsA- or CCR1 siRNA-treated cells were used. Moreover, treatment with a Galpha inhibitor pertussis toxin (PTX) or methiolynated regulated on activation, normal T cells expressed and secreted (Met-RANTES), an antagonist of CCR1, suppressed multinucleated cell formation in the bone marrow cell system. Together, these results suggest that the CCR1 signaling cascade is under the control of NFAT2 and seems to enhance the migration of differentiating osteoclasts. PMID- 16355274 TI - Degradation of the organic phase of bone by osteoclasts: a secondary role for lysosomal acidification. AB - Osteoclasts degrade bone matrix by secretion of hydrochloric acid and proteases. We studied the processes involved in the degradation of the organic matrix of bone in detail and found that lysosomal acidification is involved in this process and that MMPs are capable of degrading the organic matrix in the absence of cathepsin K. INTRODUCTION: Osteoclasts resorb bone by secretion of acid by the vacuolar H+-adenosine triphosphatase (V-ATPase) and the chloride channel ClC-7, followed by degradation of the matrix, mainly collagen type I, by cathepsin K and possibly by matrix metalloproteinases (MMPs). However, the switch from acidification to proteolysis and the exact roles of both the ion transporters and the proteinases still remain to be studied. MATERIALS AND METHODS: We isolated CD14+ monocytes from human peripheral blood from either controls or patients with autosomal dominant osteopetrosis type II (ADOII) caused by defective ClC-7 function and cultured them in the presence of RANKL and macrophage-colony stimulating factor (M-CSF) to generate osteoclasts. We decalcified cortical bovine bone slices and studied the osteoclasts with respect to morphology, markers, and degradation of the decalcified matrix in the presence of various inhibitors of osteoclast acidification and proteolysis, using normal calcified bone as a reference. RESULTS: We found that ADOII osteoclasts not only have reduced resorption of the calcified matrix, but also 40% reduced degradation of the organic phase of bone. We found that both acidification inhibitors and cathepsin K inhibitors reduced degradation of the organic matrix by 40% in normal osteoclasts, but had no effect in the ADOII osteoclasts. Furthermore, we showed that inhibition of MMPs leads to a 70% reduction in the degradation of the organic bone matrix and that MMPs and cathepsin K have additive effects. Finally, we show that osteoclastic MMPs mediate release of the carboxyterminal telopeptide of type I collagen (ICTP) fragment in the absence of cathepsin K activity, and therefore, to some extent, are able to compensate for the loss of cathepsin K activity. CONCLUSIONS: These data clearly show that osteoclastic acidification of the lysosomes plays a hitherto nonrecognized role in degradation of the organic matrix. Furthermore, these data shed light on the complicated interplay between acidification dependent and independent proteolytic processes, mediated by cathepsin K and the MMPs, respectively. PMID- 16355275 TI - Identification of multiple osteoclast precursor populations in murine bone marrow. AB - Murine BM was fractionated using a series of hematopoietic markers to characterize its osteoclast progenitor populations. We found that the early osteoclastogenic activity in total BM was recapitulated by a population of cells contained within the CD11b(-/low) CD45R- CD3- CD115high fraction. INTRODUCTION: Osteoclasts are of hematopoietic origin and they have been shown to share the same lineage as macrophages. We further characterized the phenotype of osteoclast progenitor populations in murine bone marrow (BM) by analyzing their cell surface markers. MATERIALS AND METHODS: We used fluorescence-activated cell sorting (FACS) to identify the subsets of BM cells that contained osteoclast progenitors. We fractionated BM according to several markers and cultured the sorted populations for a period of 2-6 days with macrophage-colony stimulating factor (M CSF) and RANKL. The numbers of multinucleated osteoclast-like cells (OCLs) that formed in the cultures were counted. RESULTS: We found that the CD45R- CD11b( /low) population recapitulated the early osteoclastogenic activity of total BM. In addition, although previous experiments indicated that osteoclastogenic activity was enriched within the CD45R+ population, we found that highly purified CD45R+ BM was incapable of differentiating into osteoclasts in vitro. We also found that CD45R- CD11b(high) BM cells were an inefficient source of osteoclast progenitors. However, CD11b was transiently upregulated by cells of the CD45R- CD11b(-/low) fraction early (within 24 h) during culture with M-CSF. Finally, further fractionation of BM using CD115 and CD117 showed that, as osteoclast precursor cells matured, they downregulate CD117 but remain CD115+. Curiously, pure populations of CD117- (CD115high) cells isolated fresh from BM have low osteoclastogenic activity in vitro. CONCLUSIONS: We provided a refined analysis of the precise subpopulations of murine BM that are capable of differentiating into OCLs in vitro when treated with M-CSF and RANKL. PMID- 16355276 TI - Decreased collagen organization and content are associated with reduced strength of demineralized and intact bone in the SAMP6 mouse. AB - To examine the link between bone material properties and skeletal fragility, we analyzed the mechanical, histological, biochemical, and spectroscopic properties of bones from a murine model of skeletal fragility (SAMP6). Intact bones from SAMP6 mice are weak and brittle compared with SAMR1 controls, a defect attributed to reduced strength of the bone matrix. The matrix weakness is attributed primarily to poorer organization of collagen fibers and reduced collagen content. INTRODUCTION: The contribution of age-related changes in tissue material properties to skeletal fragility is poorly understood. We previously reported that bones from SAMP6 mice are weak and brittle versus age-matched controls. Our present objectives were to use the SAMP6 mouse to assess bone material properties in a model of skeletal fragility and to relate defects in the mechanical properties of bone to the properties of demineralized bone and to the structure and organization of collagen and mineral. MATERIALS AND METHODS: Femora from 4- and 12-month-old SAMR1 (control) and SAMP6 mice were analyzed using bending and torsional mechanical testing of intact bones, tensile testing of demineralized bone, quantitative histology (including collagen fiber orientation), collagen cross-links biochemistry, and Raman spectroscopic analysis of mineral and collagen. RESULTS: Intact bones from SAMP6 mice have normal elastic properties but inferior failure properties, with 60% lower fracture energy versus SAMR1 controls. The strength defect in SAMP6 bones was associated with a 23% reduction in demineralized bone strength, which in turn was associated with poorer collagen fiber organization, lower collagen content, and higher hydroxylysine levels. However, SAMP6 have normal levels of collagen cross-links and normal apatite mineral structure. CONCLUSIONS: Bones from SAMP6 osteoporotic mice are weak and brittle because of a defect in the strength of the bone matrix. This defect is attributed primarily to poorer organization of collagen fibers and reduced collagen content. These findings highlight the role of the collagen component of the bone matrix in influencing skeletal fragility. PMID- 16355277 TI - Standardized bending and breaking test for the normal and osteoporotic metaphyseal tibias of the rat: effect of estradiol, testosterone, and raloxifene. AB - The fracture of bone plays a key role in osteoporosis. BMD measurement, however, is only an indirect parameter of this phenomenon. We therefore developed a highly sensitive three-point bending test for the metaphyseal tibias in rats to evaluate stiffness and strength. This was validated in a right-left comparison and a bioassay with soy-free food, estradiol, raloxifene, and testosterone in orchidectomized rats. INTRODUCTION: Osteoporosis becomes manifest predominantly in the metaphyseal rat tibia. The anti-osteoporotic character of substances should, therefore, be tested (mechanically) in this bone area. MATERIALS AND METHODS: We evaluated a new three-point bending test for the metaphyseal tibia in rats in a right-left trial. In an animal experiment, we studied the change of bone quality under estradiol (E)-, raloxifene (R)-, and testosterone (T) supplemented food and compared it with trabecular BMD (qCT). RESULTS: In the right-left comparison, the mean difference between the metaphyseal loads of both tibias in 37 rats was 8.43% for the maximum load (Fmax) and 6.46% for the failure load (fL). These results show the high reproducibility of the test, because they are close to the usual intraindividual difference of the two extremities. In a second experiment, four groups of 11 3-month-old male orchidectomized rats were fed with soy-free food only (C) or with the additives E, T, or R for 12 weeks. E and R were similar for Fmax and fL. There were significant differences in the stiffness (E = 406.92 N/mm versus R = 332.08 N/mm), the yield load (yL; E = 99.17 N versus R = 83.33 N), and the ratio between yL and Fmax (E = 86.33% versus R = 76.37%). T was similar to the controls concerning F(max), fL, and stiffness. There were significant differences in yL (T = 49.00N versus C = 39.5N) and the ratio between yL and Fmax (T = 64.28% versus C = 51.28%). CONCLUSIONS: Estradiol is superior to raloxifene concerning stiffness and yield load, and both are superior to testosterone. We conclude that the described three-point bending test for the metaphyseal tibia is a highly sensitive method to study hormones and substances with regard to their osteoprotective character. The precision and the low SD of the presented results are superior to the data from qCT and the calculated index of stiffness (SSI). PMID- 16355278 TI - Multiple genetic loci from CAST/EiJ chromosome 1 affect vBMD either positively or negatively in a C57BL/6J background. AB - Skeletal phenotype analyses of 10 B6.CAST-1 congenic sublines of mice have revealed evidence for the presence of three closely linked QTLs in Chr 1 that influence femoral vBMD both positively and negatively. INTRODUCTION: BMD is an important component of bone strength and a recognized predictor of risk for osteoporotic fracture. Our goal in this study was to fine map the chromosomal location of volumetric BMD (vBMD) quantitative trait loci (QTLs) in mouse distal chromosome 1 (Chr 1). MATERIALS AND METHODS: After several backcrosses of the B6.CAST-1T congenic strain, which carried the initial BMD QTL in Chr 1 with B6 mice, the N10F1 generation mice were intercrossed to obtain recombinations that yielded different regions of the QTL. Thirty-eight polymorphic markers were used to fine map the initial 1T QTL region (100-192 Mb). Different skeletal parameters were compared between the 10 sublines and B6 female mice at 16 weeks of age. A t test was used to determine the significant difference between sublines and B6 control mice, whereas one-way ANOVA and posthoc (Newman-Keuls) tests were performed to compare the phenotype between the sublines. RESULTS: Significantly higher femur vBMD was found in sublines that carried cast alleles from 100 to 169 and 172 to 185 Mb of the centromere compared with the B6 control mice (10-12%, p < 0.001). However, sublines that carried cast alleles from 185 to 192 Mb showed significantly lower femur vBMD compared with the control mice (-6%, p < 0.05). Furthermore, femur vBMD phenotype showed a negative correlation with endosteal circumference (r = -0.8, p = 0.003), and a strong correlation with cortical thickness for combined data from the 10 sublines (r = 0.97, p < 0.001). Moreover, a high correlation was found between body weight and both periosteal and endosteal circumferences for sublines carrying cast alleles from 167 to 175, 168 to 185, and 169 to 185 Mb, whereas no significant correlation was found between these parameters for sublines carrying cast alleles from 172 to 185 Mb. CONCLUSIONS: Genetic analysis using congenic sublines revealed that the initial BMD QTL on Chr 1 is a complex site with multiple loci affecting bone phenotypes, showing the value of the congenic approach in clearly identifying loci that control specific traits. PMID- 16355279 TI - Quantitative trait loci that determine BMD in C57BL/6J and 129S1/SvImJ inbred mice. AB - BMD is highly heritable; however, little is known about the genes. To identify loci controlling BMD, we conducted a QTL analysis in a (B6 x 129) F2 population of mice. We report on additional QTLs and also narrow one QTL by combining the data from multiple crosses and through haplotype analysis. INTRODUCTION: Previous studies have identified quantitative trait loci (QTL) that determine BMD in mice; however, identification of genes underlying QTLs is impeded by the large size of QTL regions. MATERIALS AND METHODS: To identify loci controlling BMD, we performed a QTL analysis of 291 (B6 x 129) F2 females. Total body and vertebral areal BMD (aBMD) were determined by peripheral DXA when mice were 20 weeks old and had consumed a high-fat diet for 14 weeks. RESULTS AND CONCLUSIONS: Two QTLs were common for both total body and vertebral aBMD: Bmd20 on chromosome (Chr) 6 (total aBMD; peak cM 26, logarithm of odds [LOD] 3.8, and vertebral aBMD; cM 32, LOD 3.6) and Bmd22 on Chr 1 (total aBMD; cM 104, LOD 2.5, and vertebral aBMD; cM 98, LOD 2.6). A QTL on Chr 10 (Bmd21, cM 68, LOD 3.0) affected total body aBMD and a QTL on Chr 7 (Bmd9, cM 44, LOD 2.7) affected vertebral aBMD. A pairwise genome-wide search did not reveal significant gene-gene interactions. Collectively, the QTLs accounted for 21.6% of total aBMD and 17.3% of vertebral aBMD of the F(2) population variances. Bmd9 was previously identified in a cross between C57BL/6J and C3H/HeJ mice, and we narrowed this QTL from 34 to 22 cM by combining the data from these crosses. By examining the Bmd9 region for conservation of ancestral alleles among the low allele strains (129S1/SvImJ and C3H/HeJ) that differed from the high allele strain (C57BL/6J), we further narrowed the region to approximately 9.9 cM, where the low allele strains share a common haplotype. Identifying the genes for these QTLs will enhance our understanding of skeletal biology. PMID- 16355280 TI - Initial characterization of PTH-related protein gene-driven lacZ expression in the mouse. AB - The PTHrP gene generates low-abundance mRNA and protein products that are not easily localized by in situ hybridization histochemistry or immunohistochemistry. We report here a PTHrP-lacZ knockin mouse in which beta-gal activity seems to provide a simple and sensitive read-out of PTHrP gene expression. INTRODUCTION: PTH-related protein (PTHrP) is widely expressed in fetal and adult tissues, typically as low-abundance mRNA and protein products that maybe difficult to localize by conventional methods. We created a PTHrP-lacZ knockin mouse as a means of surveying PTHrP gene expression in general and of identifying previously unrecognized sites of PTHrP expression. MATERIALS AND METHODS: We created a lacZ reporter construct under the control of endogenous PTHrP gene regulatory sequences. The AU-rich instability sequences in the PTHrP 3' untranslated region (UTR) were replaced with SV40 sequences, generating products with lacZ/beta gal kinetics rather than those of PTHrP. A nuclear localization sequence was not present in the construct. RESULTS: We characterized beta-galactosidase (beta-gal) activity in embryonic whole mounts and in the skeleton in young and adult animals. In embryos, we confirmed widespread PTHrP expression in many known sites and in several novel epidermal appendages (nail beds and footpads). In costal cartilage, beta-gal activity localized to the perichondrium but not the underlying chondrocytes. In the cartilaginous molds of forming long bones, beta gal activity was first evident at the proximal and distal ends. Shortly after birth, the developing secondary ossification center formed in the center of this PTHrP-rich chondrocyte population. As the secondary ossification center developed, it segregated this population into two distinct PTHrP beta-gal+ subpopulations: a subarticular subpopulation immediately subjacent to articular chondrocytes and a proliferative chondrocyte subpopulation proximal to the chondrocyte columns in the growth plate. These discrete populations remained into adulthood. beta-gal activity was not identified in osteoblasts but was present in many periosteal sites. These included simple periosteum as well as fibrous tendon insertion sites of the so-called bony and periosteal types; the beta-gal expressing cells in these sites were in the outer fibrous layer of the periosteum or its apparent equivalents at tendon insertion sites. Homozygous PTHrP-lacZ knockin mice had the expected chondrodysplastic phenotype and a much expanded region of proximal beta-gal activity in long bones, which appeared to reflect in large part the effects of feedback signaling by Indian hedgehog on proximal cell proliferation and PTHrP gene expression. CONCLUSIONS: The PTHrP-lacZ mouse seems to provide a sensitive reporter system that may prove useful as a means of studying PTHrP gene expression. PMID- 16355281 TI - Effects of sex and age on bone microstructure at the ultradistal radius: a population-based noninvasive in vivo assessment. AB - In a population-based cross-sectional study, we examined effects of sex and age on bone microstructure at the wrist using high-resolution 3-D pQCT. Compared with women, men had thicker trabeculae in young adulthood and had less microstructural damage with aging. These findings may contribute to the virtual immunity of men to age-related increases in wrist fractures. INTRODUCTION: Although changes in bone microstructure contribute to fracture risk independently of BMD, it has not heretofore been possible to assess this noninvasively in population-based studies. MATERIALS AND METHODS: We used high-resolution 3-D pQCT imaging (voxel size, 89 mum) to define, in a random sample of women (n = 324) and men (n = 278) 21-97 years of age, sex and age effects on bone microstructure at the wrist. RESULTS: Relative to young women (age, 20-29 years), young men had greater trabecular bone volume/tissue volume (BV/TV; by 26%, p = 0.001) and trabecular thickness (TbTh; by 28%, p < 0.001) but similar values for trabecular number (TbN) and trabecular separation (TbSp). Between ages 20 and 90 years, cross sectional decreases in BV/TV were similar in women (-27%) and in men (-26%), but whereas women had significant decreases in TbN (-13%) and increases in TbSp (+24%), these parameters had little net change over life in men (+7% and -2% for TbN and TbSp, respectively; p < 0.001 versus women). However, TbTh decreased to a greater extent in men (-24%) than in women (-18%; p = 0.010 versus men). CONCLUSIONS: Whereas decreases with age in trabecular BV/TV are similar in men and women, the structural basis for the decrease in trabecular volume is quite different between the sexes. Thus, over life, women undergo loss of trabeculae with an increase in TbSp, whereas men begin young adult life with thicker trabeculae and primarily sustain trabecular thinning with no net change in TbN or TbSp. Because decreases in TbN have been shown to have a much greater impact on bone strength compared with decreases in TbTh, these findings may help explain the lower life-long risk of fractures in men, and specifically, their virtual immunity to age-related increases in distal forearm fractures. PMID- 16355282 TI - Two-year clinical trial of oral alendronate versus intravenous pamidronate in children with osteogenesis imperfecta. AB - A 2-year prospective, partially randomized open-label trial comparing oral alendronate with intravenous pamidronate therapy in children with OI showed equivalence in increasing total body BMD, spine BMD, and linear growth, and decreasing bone turnover and fracture incidence. Children with mild OI had greater responses than severe OI in BMD and growth. INTRODUCTION: Bisphosphonate therapies increase BMD and may reduce fractures in children with osteogenesis imperfecta (OI). A study directly comparing oral with intravenous bisphosphonate has not been published. This clinical trial compares oral alendronate with intravenous pamidronate in children with OI using an open-label, prospective, 2 year, randomized design. MATERIALS AND METHODS: Children over the age of 3 years were stratified by bone age, pubertal stage, and type of OI and then randomized to receive oral alendronate 1 mg/kg/day in tablet form or intravenous pamidronate, 3 mg/kg/4 months. One child was assigned to pamidronate. One child randomized to intravenous pamidronate changed to oral alendronate. Eighteen children completed 12 months of therapy: nine on oral alendronate and nine on intravenous pamidronate. Primary outcome efficacy was increase in BMD. Secondary outcomes included changes in bone turnover biomarkers, fracture incidence, and growth. RESULTS: Total body and lumbar spine BMD increased, turnover markers decreased, and linear growth increased equivalently with oral and intravenous therapy. Fracture incidence showed a trend to decrease in both groups, with a significant decrease in fracture rates when the oral and intravenous groups were pooled. There were greater responses in BMD and growth in children with milder OI (type I) than those with more severe disease (types III and IV), but there were no significant effects of age or pubertal stage. CONCLUSIONS: Oral and intravenous bisphosphonate therapies are equally effective in children with OI and are particularly effective in milder forms. The oral route is highly acceptable in children and has practical advantages over the intravenous route. PMID- 16355283 TI - Common genetic variation of the low-density lipoprotein receptor-related protein 5 and 6 genes determines fracture risk in elderly white men. AB - Both LRP5 and LRP6 genes have been implicated to play a role in bone metabolism. In a large population-based study, we related common variation in both genes to bone parameters and fractures. LRP5 variation was associated to both BMD and frame size, whereas both LRP5 and 6 variations were associated with an increased fracture risk in males. INTRODUCTION: The low-density lipoprotein receptor related protein 5 (LRP5) gene has a clear role in rare BMD traits and also in normal variation in peak BMD. We examined whether common variation in LRP5 and its close homolog, LRP6, plays a role in BMD in old age and fractures, the main clinical endpoint of osteoporosis. MATERIALS AND METHODS: We analyzed four variants of LRP5 and one amino acid variant of the LRP6 gene in a large prospective population-based cohort study of elderly subjects. RESULTS AND CONCLUSIONS: In men, the LRP5 1330-valine variant was associated with decreased BMD at the lumbar spine and the femoral neck with evidence for an allele-dose effect (p = 0.001 and 0.01, respectively). The Val allele was also associated with decreased vertebral body size and femoral neck width. Haplotype analysis of studied polymorphisms did not improve the association found and suggested that the 1330 variant was driving the association. We observed a borderline significant association of the LRP6 Ile1062Val polymorphism with height and vertebral body size in males. Male carriers of the LRP5 1330-valine variant had a 60% increased risk for fragility fractures, and the LRP6 1062-valine allele also conferred a 60% higher risk. Carriers of both the risk alleles of LRP5 and 6 had a 140% (p = 0.004) higher risk compared with noncarriers of both risk alleles and accounted for 10% of the fractures in males. The fracture risks were independent of age, height, weight, and BMD. In women, all of these associations were weaker and less consistent compared with men. The polymorphisms that were found associated were both situated in potentially important domains of the receptor and show considerable evolutionary conservation, which is evidence for functional importance of these residues. PMID- 16355284 TI - Large-scale population-based study shows no evidence of association between common polymorphism of the VDR gene and BMD in British women. AB - The VDR is a candidate gene for osteoporosis. Here we studied five common polymorphisms of VDR in relation to calcium intake and vitamin D status in a population-based cohort of 3100 British women, but found no significant association with bone mass, bone loss, or fracture. INTRODUCTION: Population studies of vitamin D receptor (VDR) polymorphisms have produced conflicting results. We performed a comprehensive study dealing with all potential confounders in a large population to determine whether polymorphisms in the VDR gene influence bone health. MATERIALS AND METHODS: We studied 3100 women (50-63 years old) with bone markers, 25-hydroxyvitamin D, calcium, PTH, diet, and physical activity collected in 1998-2000. BMD was measured in 1990-1994 and 1998 2000. Fracture prevalence was assessed in 2002. Women were genotyped for five polymorphisms in the VDR gene: Cdx-2, Fok1, Bsm1, Apa1, and Taq1. The relationship between VDR and BMD, and interactions between VDR genotype, dietary calcium, and 25-hydroxyvitamin D, were examined using analysis of covariance. RESULTS: Compared with carriers of the G allele, homozygotes for the rare Cdx-2 A polymorphism (n = 136) had less bone loss (-0.5 +/- 1.2 versus -0.7 +/- 1.0%/year [SD]; p = 0.01) and lower PTH (3.0 +/- 1.6 versus 3.4 +/- 2.0 pM; p = 0.03) despite similar vitamin D status. The association was not significant after correction for multiple testing or adjustment for confounders. At low calcium intakes, AA homozygotes had greater femoral neck (FN) BMD compared with carriers of the G allele, but at higher calcium intakes, the association was reversed. At low calcium intake, homozygotes for the b allele of Bsm1 had greater BMD compared with carriers of the B allele, but at higher calcium intakes, there was no difference. Similar results were seen for the Taq1 polymorphism. There was no evidence of gene-nutrient interaction when adjusted for body weight. No interactions between genotypes and vitamin D status on BMD were observed. CONCLUSIONS: VDR does not seem to influence BMD or bone turnover in early postmenopausal white women with adequate calcium intake. Gene-nutrient interactions on BMD may be an indirect consequence of interactions between genotype and calcium intake on weight. PMID- 16355285 TI - Loop diuretics increase bone turnover and decrease BMD in osteopenic postmenopausal women: results from a randomized controlled study with bumetanide. AB - To study effects of loop diuretics on bone, 87 women were randomized to 1 year of treatment with bumetanide or placebo. Compared with placebo, bumetanide decreased BMD by 2% at the total hip and by 1.4% at the whole body. Levels of biochemical bone markers were lower in the placebo than in the bumetanide group. Thus, treatment with loop diuretics affects bone metabolism. INTRODUCTION: Loop diuretics may affect bone because they increase the renal calcium excretion and alters the diurnal rhythm of plasma PTH levels. We studied the effects of 1 year of treatment with the loop diuretic bumetanide on bone metabolism. MATERIALS AND METHODS: In a double-blinded design, 87 healthy postmenopausal women with osteopenia were randomized to 1-year bumetanide treatment 2 mg/day or placebo. BMD, plasma levels of calcitropic hormones, and biochemical bone markers were measured at baseline, after 1 year of treatment (week 52), and 6 months after withdrawal of treatment (week 78). Calcium (800 mg/day) and vitamin D (10 microg/day) were administered to all participants during the entire 1.5-year study period. RESULTS: Compared with placebo, urinary calcium (+17%) and plasma PTH levels (+9%) increased significantly in response to bumetanide. After 1 year of treatment, BMD in the bumetanide compared with the placebo group was significantly decreased by 2% at the total hip and ultradistal forearm and by 1.4% at the whole body. In addition, levels of biochemical markers of bone turnover differed significantly (approximately 20%) between groups, with lower levels in the placebo than in the bumetanide group. Six months after the end of treatment, the effects of bumetanide were weakening. CONCLUSIONS: Treatment with loop diuretics affects bone turnover and decreases BMD. Further studies may reveal whether loop diuretics should be considered as a risk factor for fracture. PMID- 16355286 TI - Incidence of primary hyperparathyroidism in Rochester, Minnesota, 1993-2001: an update on the changing epidemiology of the disease. AB - We updated the incidence of primary hyperparathyroidism in Rochester, Minnesota. The lower rates previously noted persisted, whereas parathyroidectomies at our institution remained high. These data suggest an etiologic factor may be responsible for the peak incidence in the 1970s. INTRODUCTION: Automated serum calcium measurements were associated with a dramatic rise in primary hyperparathyroidism in the early 1970s, but a progressive decline in the incidence thereafter was unexpected and suggested a fundamental change in the epidemiology of the disease. Our objective was to evaluate trends in the incidence of primary hyperparathyroidism since 1992. MATERIALS AND METHODS: In this population-based descriptive study, Rochester, MN, residents who met defined diagnostic criteria for primary hyperparathyroidism from January 1993 through December 2001 were identified through the medical record linkage system of the Rochester Epidemiology Project and the Mayo Clinic Laboratory Information System. Changes in incidence were evaluated by Poisson regression. RESULTS: Altogether, 136 Rochester residents (94 women and 42 men) were newly identified with primary hyperparathyroidism in 1993-2001. Their mean age was 56 years, and 93% had definite disease. The overall age- and sex-adjusted (to 2000 U.S. whites) rate during this period was 21.6 per 100,000 person-years, which was less than the annual rate of 29.1 per 100,000 observed in 1983-1992 and 82.5 per 100,000 in July 1974-1982. Although community incidence declined, the number of parathyroidectomies performed at our institution increased during the same period. Serum calcium was deleted from the automated chemistry panel in June 1996, but most subjects remained asymptomatic at diagnosis (95%) with mild hypercalcemia. The majority of subjects were observed without parathyroid surgery (75%), and there was minimal impact on patient management from the 1990 NIH consensus conference on asymptomatic primary hyperparathyroidism. CONCLUSIONS: The lower incidence of primary hyperparathyroidism noted through 1992 has persisted in our community through 2001, whereas parathyroidectomies at our institution remained high. These data suggest that some underlying etiologic factor, in addition to the introduction of automated serum calcium testing, may have been responsible for the peak incidence in the 1970s. PMID- 16355287 TI - Bone remodeling: biochemical markers or bone biopsy? PMID- 16355289 TI - Academic specialist education in headache medicine. Next move for the headache community. PMID- 16355290 TI - The pathophysiology of migraine: year 2005. AB - Migraine is a complex patholophysiology in which both central and peripheral components of the trigeminal pain pathway probably play a significant role, both in the symptoms and signs of the attack and in the mechanisms of action of antimigraine compounds, such as triptans, which constitute the most important therapy for aborting migraine pain and possess several mechanisms on 5-HT receptor-mediated actions. The experimental neurogenic inflammation model represents a simple procedure to obtain preliminary information on well characterized receptortargeted drugs. The apparent paradox observed with certain drugs that are shown to be effective in this model but not in clinical trials offers the opportunity to better manipulate structure-activity to obtain the best pharmacological profile using an array of experimental models. The observation that nitric oxide donors induce migraine-like pain in migraineours and that nitric oxide plays a pivotal role in the control of several functions in the central nervous system, has prompted the use of such molecules for better understanding the pathophysiology of migraine attacks. A link between central and peripheral components of the trigeminal pain pathway is provided by the observation that cortical spreading depression in the rat activates trigeminovascular afferents and induces a series of cortical meningeal and brainstem events consistent with the development of headache. Studies in humans support the hypothesis that cortical spreading depression underlies migraine.aura. Therefore, tt is possible that visual, motor or sensory aura might be responsible for the generation of the pain through the above mechanisms. PMID- 16355291 TI - Patients' preference for triptans and other medications as a tool for assessing the efficacy of acute treatments for migraine. AB - Oral triptans are effective and well tolerated acute treatments for migraine, but clinical differences between them are small and difficult to measure in conventional clinical trials. Patient preference assesses a global measure of efficacy and tolerability, and may be a more sensitive means of distinguishing between these drugs. In a series of studies, patients consistently expressed a clear preference for triptans over their usual non-triptan acute medications, e.g., analgesics and ergotamine. Direct comparator studies of patient preference with oral triptans showed that patients could distinguish between different triptans, and between different formulations of the same triptan. Patients could even distinguish between the three oral doses of sumatriptan. The most frequently provided reasons for preference were speed of response and overall effectiveness. Patient preference is a sensitive clinical trial endpoint and physicians should consider using it when reviewing the efficacy of acute migraine medications. PMID- 16355292 TI - Nosology and treatment of primary headache in a Swiss headache clinic. AB - We assessed demographics, diagnoses, course, severity, impact and treatment of primary headache outpatients from records in the Headache and Pain Clinic, Neurological Department, Zurich University Hospital. All outpatients seen from 1996 to 1998 for migraine, tension-type headache, and both, were included. Diagnoses, drug, physical and alternative treatments before and after referral were listed. Descriptive statistics were used for differences between the general population and this sample, the diagnoses, and treatments. The coexistence of migraine and tension-type headache, and the high frequencies of headache days would have excluded most migraine patients from typical drug trials: at best, only one third were eligible. The socioeconomic impact of combined and difficult syndromes calls for comprehensive management beyond simple treatment with instant relief drugs. The diagnostic and therapeutic practices of referring physicians exposed a deficit of information on headache, and a need for relevant education. PMID- 16355293 TI - Demographic, clinical and comorbidity data in a large sample of 1,147 patients with migraine in Mexico City. AB - The objective was to identify the sociodemographic and clinical characteristics of a large sample of patients with migraine in Mexico City. This cross-sectional study was performed in two tertiary centers in Mexico City and affiliated hospitals. We evaluated the presence of migraine through a standardised interview according to the criteria of the International Headache Society. We studied 1,147 patients. The mean age was 37.1+/-13.6 (6-77) years. Nine hundred and twenty one patients were female (80%). The age of onset of migraine was 19.4+/-10.3 (1-69) years. Six hundred and four patients had migraine with aura (53%) and 543 without aura (47%). The female/male ratio was 4:1. One hundred and forty-seven patients had cardiovascular problems (13%), 72 had neurological problems (6%), 233 had gastrointestinal problems (20%) and 323 had psychiatric problems (28%). In this study we described the clinical characteristics of a large sample of patients with migraine in Mexico City. Our sample has similar characteristics to other countries. PMID- 16355294 TI - Recurrent and chronic headaches in children below 6 years of age. AB - The objective was to determine the frequency of headache subtypes, according to International Headache Society (IHS) criteria, in a population of children below 6 years visiting a Center for Diagnosis and Treatment of Headache in Youth. Medical records of the children below 6 years at their first visit, admitted for headache between 1997 and 2003, were studied. Headache was classified according to the IHS criteria 2004. Children with less than three headache attacks or less than 15 days of daily headache were excluded. We found 1598 medical records of children who visited our Headache Center in the study period. One hundred and five (6.5%) were children younger than 6 years. The mean age at the first medical control was 4.8+/-1.3 years (range 17-71 months). There were 59 males (56.1%) and 46 females (43.9%). The mean age at onset of headaches was 4.3 years (range 14-69 months). According to the IHS criteria we found 37 cases (35.2%) with migraine, 19 cases (18%) with episodic tension headache, 5 cases (4.8%) with chronic daily headache, 13 cases (12.4%) with primary stabbing headache, 18 cases (17.1%) with post-traumatic headache, 7 cases (6.6%) with other non-dangerous secondary headaches (otorhinolaryngological diseases, post-infectious headaches), 3 cases (2.85%) with dangerous headaches (Arnold-Chiari type 1 malformation, brain tumour) and 9 cases (8.6%) with unclassifiable headaches. Six children (5.7%) reported more than one headache subtype. The prevalence of dangerous headaches was higher than those in school age (chi(2)=4.70, p<0.05). Our study shows some differences in headaches in this population vs. school children. In fact at this age migraine is the most common headache, but we also found an increase of secondary causes among the chronic/recurrent and daily headaches, especially posttraumatic disorders and potentially dangerous headaches. Finally our study shows the highest prevalence of the idiopathic stabbing headache in pre-school children in comparison with other ages. PMID- 16355295 TI - Intramuscular tramadol vs. diclofenac sodium for the treatment of acute migraine attacks in emergency department: a prospective, randomised, double-blind study. AB - The aim of this prospective, randomised, double-blind study was to evaluate the efficacy of intramuscular (IM) tramadol 100 mg in emergency department treatment of acute migraine attack and to compare it with that of IM diclofenac sodium 75 mg. Forty patients who were admitted to our emergency department with acute migraine attack according to the International Headache Society criteria were included in the study. Patients were randomised to receive either tramadol 100 mg (n=20) or diclofenac sodium 75 mg (n=20) intramuscularly. Patients rated their pain on a four-point verbal scale (0=none, 1=mild, 2=moderate, 3=severe) at the beginning of the trial and at 30, 60, 90 and 120 min. At each time interval, severity of associated symptoms were also questioned and recorded. Global evaluation of the drugs by patients and doctors were also recorded. Patients were also asked if they would prefer the same injection in future visits. Any adverse events, whether related to the drug or not, were also recorded. Patients were followed up by telephone 48 h later to check for any headache recurrence. Two hour pain response rate, which was the primary endpoint, was 80% for both tramadol and diclofenac groups. There were no statistically significant differences among groups in terms of 48-h pain response, rescue treatment, associated symptoms' response, headache recurrence and adverse event rates. Fifteen (75%) patients in the tramadol group and 16 (80%) patients in the diclofenac group stated that they may prefer the same agent for future admissions. In selected patients, tramadol 100 mg IM may be an effective and reliable alternative treatment choice in acute migraine attacks. PMID- 16355296 TI - Non-hypothalamic cluster headache: the role of the greater occipital nerve in cluster headache pathogenesis. AB - Cluster headache is marked by its circadian rhythmicity and the hypothalamus appears to have a significant influence over cluster pathogenesis. However, as not all cluster patients present in the same manner and not all respond to the same combination of medications, there is likely a nonhypothalamic form of cluster headache. A patient is presented who began to develop cluster headaches after receiving bilateral greater occipital nerve (GON) blockade. His headaches fit the IHS criteria for cluster headache but had some irregularities including frequent side shifting of pain, irregular duration and time of onset and the ability of the patient to sit completely still during a headache without any sense of agitation. This article will suggest that some forms of cluster headache are not primarily hypothalamic influenced and that the GON may play a significant role in cluster pathogenesis in some individuals. PMID- 16355298 TI - Headache attributed to unruptured saccular aneurysm, mimicking hemicrania continua. AB - Unruptured cerebral arterial aneurysms most often remain asymptomatic, but they may cause headache or other symptoms or signs. We describe herewith a case of headache attributed to an unruptured internal carotid artery aneurysm, clearly mimicking the phenotype of hemicrania continua. Potential pathophysiological explanations and recommendations for recognition of similar cases are discussed. PMID- 16355297 TI - A case of a GH-producing pituitary adenoma associated with a unilateral headache with autonomic signs. AB - A 66-year-old man suffered from a drug-resistant, left-sided headache with autonomic signs, triggered by the supine position. The acromegalic facies initially suggested a possible increase in basal plasma levels of GH, but routine haematological controls excluded abnormal values of GH. Cerebral and facial CT scan and MRI did not detect any alterations in the nasal sinuses, except for a mucous cyst. Surgical ablation of the cyst did not alleviate the pain. Further endocrinological the pain. Further endocrinological tests demonstrated an increase of IGF-1 (somatomedin C), and another MRI scan of the sellar region confirmed the presence of a pituitary macroadenoma on the left paramedian side. After an initial improvement of the symptomatology due to trans-sphenoidal ablation of a benign GH-producing macroadenoma, the headache worsened again. Pain was well correlated with the increased plasma levels of IGF-1. The patient died suddenly for myocardial infarct. PMID- 16355299 TI - Training on the International Classification of Functioning, Disability and Health (ICF): the ICF-DIN Basic and the ICF-DIN Advanced Course developed by the Disability Italian Network. AB - The objective is to present training on the International Classification of Functioning, Disability and Health (ICF) prepared by the Disability Italian Network (DIN) and to present strategies of ICF dissemination in Italy. A description of DIN's training methodology, prepared in collaboration with World Health Organization (WHO) experts, is provided within its practical applications in health, labour, rehabilitation and statistical sectors. The ICF-DIN Basic Course is eight hours long and focuses on ICF basic principles, structure and application in different settings. The ICF-DIN Advanced Course, three days long followed by three months of distance learning, assumes Basic Course completion, and focuses also on ICF-checklist's coding and WHO-DAS II administration. The first training courses' outcomes, held in Italy and addressed to health, social and labour professionals, are provided. The feedback received by participants at the end of the courses showed that the main mistake they made was to consider ICF as an assessment instrument. The ICF-DIN training course was crucial in explaining the correct use of the ICF as a classification and to show its impact and usefulness on daily practice, particularly in multidisciplinary teams. The ICF-DIN courses already carried out in Italy show that this teaching methodology teaches how to avoid incomplete applications, simplification and misunderstanding of ICF's complexity. PMID- 16355300 TI - [Brain perfusion disturbances in patients with vertigo -- a study with SPECT]. AB - BACKGROUND AND PURPOSE: The aim of our study was to evaluate brain perfusion in patients with vertigo using the SPECT technique. METHODS: The study involved a group of 32 patients and was performed in the Neurological Department of the Medical University in Bialystok. Patients with vertigo of peripheral origin like middle ear pathology were excluded from the study. Tomographic pictures were taken with Nucline X-Ring camera after administration of Tc99m-ECD. Perfusion maps were estimated by qualitative and semi-quantitative methods. RESULTS: In 8 patients (25%) perfusion maps were normal in the hemispheres, cerebellum and subcortical structures. In 10 patients (31.2%) there was a substantial decrease in perfusion in the left temporal region, in 8 patients (25%) -- hypoperfusion was seen in the right temporal region. In 4 patients (12.5%) there was a substantial decrease in perfusion in the cerebellum, in two persons -- in the frontal lobes. CONCLUSIONS: The results obtained so far confirm the major role of ischemia in etiology of the central origin vertigo and balance disturbances. It involves not only the brainstem and cerebellar structures, but the temporal lobes as well. The test has also proved that the qualitative and semi-quantitative methods of assessing brain perfusion with a SPECT are satisfactory in diagnostics of vertigo. PMID- 16355301 TI - [The role of environmental factors in Parkinson's disease may depend on disease onset age]. AB - BACKGROUND AND PURPOSE: Various factors are suspected to participate in PD onset and include environment-related factors and workplace exposure to pesticides, metals and hydrocarbons. Nevertheless, results of epidemiological research are inconsistent. Some authors emphasize hydrocarbons exposure to younger patients. Our aim was to compare PD risk factors to onset age. MATERIAL AND METHODS: Of 174 patients with idiopathic PD, without dementia, two subgroups were isolated: 65 patients with early onset PD (EOPD) below 50 (n=65, age 52.8+/-7.6 years, onset 42.8+/-5.3 years) and 109 patients with late onset (LOPD) above 50 (n=109, age 67.8+/-7.0, onset 60.8+/-6.7 years). Various environmental factors reported in literature were analyzed. RESULTS: The univariate analysis showed that factors significantly predisposing to EOPD are vocational education (OR 3.24, 95%CI 1.50 7.00, p<0.003), smoking (OR 1.94, 95%CI 1.02-3.69, p<0.05), well water consumption at 20-40 (OR 2.77, 95%CI 1.31-5.86, p<0.008), and after 40 (OR 4.84, 95%CI 1.95-11.99, p<0.0007), side-effects following exposure to paints (OR 2.26, 95%CI 1.10-4.66, p<0.03) and exposure to solvents (OR 1.98, 95%CI 0.96-4.07, p<0.07) on borderline significance. Drinking well water both between 20-40 and after 40 involved a substantial increase in EOPD (OR 6.57, 95%CI 2.43-17.75, p<0.0002). Education only at a primary level proved to be protective against EOPD (OR 0.20, 95%CI 0.07-0.55, p<0.002). The multivariate logistic regression model demonstrated that independent EOPD risk factors are smoking (OR 2.20, 95%CI 1.07 4.53, p<0.04) and well water consumption both between 20-40 and after 40 (OR 8.29, 95%CI 2.73-25.23, p<0.0002), whilst the independent protective factor is education only at a primary level (OR 0.17, 95%CI 0.05-0.53, p<0.003). CONCLUSIONS: Our research demonstrated that a number of independent environmental factors significantly affect the risk of PD onset at younger ages. Presumably, some of the observed differences in the results of research of various authors into PD risk factors may be caused by ignoring onset age within the researched patients. PMID- 16355302 TI - [Motor-evoked potentials in patients with diabetes mellitus type 1]. AB - BACKGROUND AND PURPOSE: The aim of the study was to assess the function of the central motor pathway in young patients with diabetes mellitus type 1 by use of transcranial and paravertebral magnetic stimulation. MATERIALS AND METHODS: MEPs were recorded in 68 young patients (25+/-5.69 years), with diabetes mellitus type 1, from muscles: abductor digiti minimi and abductor hallucis (AH). Central motor conduction time (CMCT) was calculated by subtracting cortical latency (CL) after transcranial stimulation from the motor nerve conduction time (MNCT) after paravertebral stimulation. The obtained results were compared with normative data from the group of 36 healthy volunteers, matched for age and height. Statistical comparison of CMCT between diabetic and control groups was performed. RESULTS: There were no significant differences between the diabetics and control means of CMCT. Also, we were unable to elicit the MEPs cortically from AH muscle in 19 (27.9%) of diabetic patients and only in 3 (8.3%) controls. CONCLUSION: CMCT is normal in patients below 40 years of age, in whom the MEPs after transcranial stimulation can be elicited. Lack of MEPs in lower limb muscles following transcranial stimulation in almost 30% of patients in the presence of MEPs in upper limbs may indirectly suggest the dysfunction of central motor conduction in those cases. PMID- 16355303 TI - [GEOS (Gabitril Epilepsy Observational Study) -- Polish results of the international study of tiagabine in partial epilepsy]. AB - BACKGROUND AND PURPOSE: Tiagabine (TGB) is a new antiepileptic drug with a unique mechanism of action, with efficacy and tolerance confirmed in many randomized, placebo-controlled add-on trials in patients with partial epilepsy. The main aim of the international GEOS study (Gabitril Epilepsy Observational Study) was to analyze the efficacy and safety of TGB as an add-on therapy in everyday clinical practice. MATERIAL AND METHODS: 260 patients from Poland (1227 in the whole study) were enrolled (mean age 35.0+/-13.7 years). 53% had symptomatic epilepsy, 25% idiopathic, 12% cryptogenic, in remaining 10% of patients the etiology of the epileptic syndrome had not been determined. 69% of patients had complex partial seizures (CPS), 60% of patients had secondary generalized tonic clonic seizures (SGTCS) and 32% of patients had simple partial seizures (SPS). The patients were observed for 12 months. RESULTS: The full study was completed by 201 (76%) of patients. The mean TGB dose received at the end of the study was 31,3+/-12 mg/day. At the end of the study, i. e. after 12 months, 75% patients with SP, 67% patients with SGTCS and 48% patients with CPS were free of seizures. 94% of patients with SPS, 92% patients with SGTCS and 91% of patients with CPS had 50% or more reduction in seizures versus baseline. TGB was well-tolerated, adverse events appeared in 14% of patients and had mild or moderate severity. CONCLUSIONS: The obtained results confirmed that TGB was effective and well tolerated in everyday clinical practice when given as add-on therapy to patients with partial epilepsy. PMID- 16355304 TI - [Neuropsychological assessment in the diagnosis and differential diagnosis of fronto-temporal dementia]. AB - Clinical diagnosis of fronto-temporal dementia (FTD) and its rare variants (semantic dementia and primary progressive aphasia) represents a special challenge bearing in mind its difficulties and is important due to therapeutic differences with the most common Alzheimer's dementia. Of note, cholinesterase inhibitors commonly prescribed in Alzheimer's disease are not effective in FTD and a novel drug, memantine, has not been evaluated as yet. Neuropsychological evaluation is an important add-on to neurological, psychiatric and neuroimaging assessments in the clinical diagnosis of FTD. In this paper, we have described characteristics of neuropsychological deficits observed in FTD (such as attention, language, visuospatial and memory impairments) as well as tools used (tests) in clinical practice. Special attention is paid to the utility of neuropsychological examination in the differential diagnosis of FTD versus Alzheimer's disease, vascular dementia and other, less common forms of dementia. PMID- 16355305 TI - [The effect of MDR1 gene polymorphism in the pathogenesis and the treatment of drug-resistant epilepsy]. AB - Recent data indicate the possibility of P-glycoprotein involvement in drug resistance in patients diagnosed with epilepsia. It was demonstrated that P glycoprotein is expressed in the endothelial cells of the blood-brain barrier, and in neurons and glial cells isolated from the epileptogenic brain tissue. The glycoprotein functions as an efflux pump, thus limiting penetration of antiepileptic drugs (phenytoin, carbamazepine, phenobarbital, gabapentin, felbamate, topiramate, lamotrigine) to the site of action. A naturally occurring MDR1 polymorphism has been described and correlated with potential clinical effects. The C3435T polymorphism was found to significantly correlate with the function of MDR1 and the expression of P-glycoprotein. This polymorphism consists of a C-to-T exchange at position 3435 in exon 26 of the MDR1 gene. Individuals with the TT genotype had significantly lower P-glycoprotein expression than those with the CC and CT genotype. Because C3435T does not change the amino acid sequence and is not located at a promotor position in the MDR1 gene, it is unlikely that this polymorphism directly influences P-glycoprotein expression. However, a strong association between the C3435T and G2677 (A, T) allele was revealed. Since G2677 (A, T) in exon 21 is a missense mutation, it is likely to be causative for differences in P-glycoprotein expression. Finding out the relationship between MDR-1 gen polymorphism and drug-resistant epilepsia may lead to the effective treatment of epilepsia by application of P-glycoprotein inhibitors. PMID- 16355306 TI - [Lactoferrin in the central nervous system]. AB - Lactoferrin (LF) is a protein secreted by the tissues of ectodermal origin. Its structure is similar to transferrin. LF appeared to be multifunctional, but its main functions are connected with the natural defense system of mammals. The biological role and origin of LF within brain in normal and disease processes are as yet uncharted. LF expression is greatly upregulated during neurodegenerative disorders and in elderly brains. LF may exert an antiinflammatory function via its inhibitory effect on hydroxyl radical formation. By antioxydative properties, LF prevents DNA damage and consequently tumor formation in the CNS. Moreover, LF specifically transactivates the p53 tumor suppressor gene. LF suppresses distress perception via opioid mediated mechanism and prevents a decrease of the immune system activity caused by psychosocial stress. Furthermore, LF possibly modulates behavior in man and in animals. PMID- 16355307 TI - [Comparison of the safety of the medicinal product in the European Union and the United States, tolcapone (Tasmar) -- COMT inhibitor as the analyzed example]. AB - The authors present regulatory procedures and pharmaco-vigilance systems obligatory in the European Union and the United States. The post-approval procedures in the European Union and the United States in case of serious drug reaction (urgent safety restriction) are discussed. Worldwide implementations of the post-approval safety procedure for the tolcapone case are analyzed. Some practical information concerning tolcapone and regulatory procedures obligatory in the European Union and the United States is included. PMID- 16355308 TI - [Research advances in molecular genetics of epilepsies]. AB - Last years brought a large number of papers shedding light on the molecular and genetic background of epilepsy. The aim of the present work was to review recent literature concerning this issue. In the first section, the authors presented biochemical and functional characteristics of receptors and ion channels. Mutations in their genes can lead to epileptic seizure development. Particularly, the role of GABA receptors as well as voltage-gated sodium, calcium and potassium channels in epilepsy pathogenesis was discussed. The second part of the work focuses on clinical implications of these receptors and channels disturbances. Epileptic syndromes with proved genetic defect are presented on the basis of recent literature and OMIM (Online Mendelian Inheritance in Man) database. The authors point out that reviewed results show vast heterogeneity of many known epileptic syndromes. On the other hand, defects in the same locus can result in heterogeneous phenotypes. This can indicate multifactorial pathogenesis of epilepsy and implicate further revision of epilepsies and epileptic seizures classification. PMID- 16355309 TI - [Suprascapular nerve entrapment. A case report]. AB - We present a case of a 35 year-old man, an office employee, who got ill when working hard building his own house. At the same time shoulder pain and fever occurred, the latter due to the tooth apical abscess. Several months later atrophy of the left supraspinatus and infraspinatus muscles was found. The imaging study (MRI, USG) did not show changes which might affect the suprascapular nerve. Electromyography of the left infraspinatus and supraspinatus muscles revealed fibrillation and positive sharp waves. Distal motor latency to the infraspinatus and supraspinatus muscles, examined by electroneurography, was normal. Mechanical irritation of the spinoglenoid notch is only one of possible mechanisms of the suprascpular nerve impairment in this case. The second is the inflammatory process. Suprascapular nerve impairment does not play an important role in pathogenesis of shoulder pain, but probably it is taken too rarely into consideration in differential diagnosis of shoulder pain. PMID- 16355310 TI - [A sixty-eight-year-old man with giant intracranial arteriovenous malformation and right-sided hemicrania -- a case report]. AB - Arteriovenous malformations (AVMs) are the most dangerous congenital vascular malformations. Intracranial AVMs occur in about 0.1 percent of the population and account for 1 to 2 percent of all strokes. The angioarchitecture of AVMs consists of direct arterial to venous connections without an intervening capillary network. The main symptom reported by half of all patients with arteriovenous malformations is an episodic headache. The headache is associated with nausea and vomiting and due to this resembles the hemicrania or cluster headache attacks. Although cerebral angiography is considered as the gold standard for the diagnosis, planned treatment and after treatment observation, AVMs are usually identified with advanced visual imaging techniques such as magnetic resonance imaging, computed tomography, and computed angiotomography. The treatment options consist of surgery, radiosurgery and endovascular embolization. We report a case of a 68-year-old man with giant intracranial arteriovenous malformation and right sided hemicrania. PMID- 16355311 TI - [Mental disorders in a female patient with probable Creutzfeldt-Jakob disease]. AB - The paper presents an overview of the current knowledge about the etiology, classification of Creutzfeldt-Jakob disease, abnormalities in the results of the EEG, MR and laboratory examinations in patients with this disease. The diagnostic value of the CSF examination for presence of protein 14-3-3 is underlined. The article is based on both Polish and foreign literature, describing mainly the diagnostics of CJD. The case of a female patient with dementia, mental disorders and neurological symptoms in the course of probable CJD, who was hospitalized at the Psychogeriatric Department of the Neuropsychiatric Hospital in Lublin is described. PMID- 16355312 TI - [Pneumocephalus after shunt in a patient with hydrocephalus. A case report and review of the literature]. AB - Pneumocephalus in hydrocephalic shunted patients is very rare. So far only 46 cases have been described in the literature. A case of pneumocephalus in a patient with shunted hydrocephalus, and the diagnostic and therapeutic problems were described. A 38-year-old woman suffered from a subacute headache, dizziness and bradyphrenia. Eight years ago, the ventriculo-atrial shunt was implanted due to idiopathic hydrocephalus, and the shunt was changed 2 months before the present admission (ventriculo-peritoneal valve). CT and MRI revealed pneumocephalus, but neither clinical nor radiological symptoms of the CSF fistula were found. After temporary ligation of a distal catheter, the anterior fossa and a sellar area were directly repaired via right fronto-temporal craniotomy, with a good outcome. In the case of pneumocephalus in hydrocephalic shunted patients, the direct closure of the skull base fistula via craniotomy combined with restoration of the proper shunt function is usually the most effective treatment. If the site of the fistula can not be established, dural cranioplasty in the most likely site of the fistula is recommended. PMID- 16355313 TI - [Cannabinoids in multiple sclerosis -- therapeutically reasonable?]. PMID- 16355314 TI - [Recent advances in pathogenesis and therapy of multiple sclerosis]. AB - In this article, recent advances in the research on pathogenesis and therapy of multiple sclerosis (MS) will be summarized. New evidence from clinical studies, imaging, histopathology and experimental models are discussed with a focus on neurodegenerative aspects and evidence from recent therapeutic studies. During the last decade, important advances in immunotherapy have been achieved, which proved especially useful for patients with relapsing remitting MS. The introduction of interferons and glatiramer acetate into MS therapy often leads to a stabilization of the disease course if administered adequately and early. The pathogenetic insights presented here may open new avenues for innovative immunomodulatory approaches and lead to an individualized MS therapy in the future. Neuroprotective treatment strategies aim at the protection of glial and neuronal cells. PMID- 16355315 TI - [Motivational interviewing for patients with comorbid schizophrenia and substance abuse disorders: a review]. AB - Patients with schizophrenia and substance abuse disorders [dually diagnosed patients (DD)] show an unfavourable course of the illness and little interest in participating on specific integrated treatment programmes. Motivational interviewing (MI) has been shown to be effective among other substance abuse disorders and it aims to enhance intrinsic motivation to change problem behaviour. MI has been adapted for DD. The present paper reviews the empirical evidence for the efficacy of MI in DD. A search in the databases MEDLINE, EMBASE, PsycINFO was conducted and the methodological quality of the identified trials was assessed according to the Cochrane Collaboration and to the JADAD Scale. We identified 4 randomised studies with a total of 346 participants, in which MI interventions of 1 to 3 sessions were compared with various control conditions over a follow-up period of up to 6 months. With regard to the main outcome measures "subsequent participation at integrated treatment programme" (1 x positive, 2 x negative) and "substance use" (1 x positive, 1 x negative,) the studies gained contradictory results. In all 4 studies, there were relevant general methodological limitations (randomisation, blindness of raters, description of the reasons for drop-outs) and specific methodological shortcomings (sample size and sample homogenity, numbers of MI sessions, assessment of motivational status). Hence, at present the evidence for supporting MI in DD is not clear. This may be due to the methodological problems mentioned above or it may be that there is, in fact, no effect. Therefore, there is an urgent need for further research of MI in DD. PMID- 16355316 TI - [The situation of siblings of mentally ill people]. AB - OBJECTIVE: The purpose of this article is to provide an overview of the current research regarding the situation of mentally ill persons. METHODS: The relevant literature was identified by means of a computerized MEDLINE search on the years 1996 - 2004 with the key words "sibling" and "mentally ill"/"chronically ill"/"psychiatric disorder" (631 articles). Also the related articles in the MEDLINE search were viewed. Finally 170 articles of journals and books were included in the review. RESULTS: The situation of siblings of psychiatric ill has manifold aspects, which can be classified into the following categories: Burdens due to the relationship to the ill sibling, burdens because of the relations to the shared parents and other family members, and burdens in the daily life of the siblings due to the illness of his/her sibling. On the other side there are rewards and positive developments in their life because of the illness of the sibling. Personal variables like sex, family size and age at onset of the illness are an influence on the experience of the sibling. CONCLUSION: The revealed burdens of the affected siblings require necessary answers by political representatives, professionals in and outside a clinic, and the involved families. PMID- 16355317 TI - [Neuroborreliosis and diphasic meningoencephalitis -- common features and differences]. PMID- 16355318 TI - Challenges in the management of serious infections. PMID- 16355319 TI - Trends in antimicrobial resistance in health care-associated pathogens and effect on treatment. AB - Antimicrobial resistance in health care-associated pathogens is a growing concern for health care and for public health. A recent shift in the epidemiological profile of methicillin-resistant Staphylococcus aureus has resulted not only in health care-associated infections but now, also, in community-associated infections. Reports have described S. aureus exhibiting decreased susceptibility and, even, resistance to vancomycin. The rate of vancomycin resistance among enterococci may be leveling; however, vancomycin resistance in Enterococcus faecium has already achieved high levels. Multidrug resistance in Pseudomonas aeruginosa is increasing, and carbapenem-resistant Klebsiella strains are emerging. Acinetobacter species cause a minority of health care-associated pneumonias due to gram-negative organisms, but a growing proportion is resistant to carbapenems and third-generation cephalosporins. Recent increases in the frequency and severity of Clostridium difficile-associated illness are associated with the emergence of a hypervirulent C. difficile strain with increased resistance to the fluoroquinolones. The presence of these and other resistant organisms in health care facilities limits the number of effective antimicrobials available for treatment. PMID- 16355320 TI - Use of broad-spectrum antimicrobials for the treatment of pneumonia in seriously ill patients: maximizing clinical outcomes and minimizing selection of resistant organisms. AB - Among various risk factors for death among critically ill patients with serious infection, inappropriate antimicrobial therapy is an important factor that clinicians can modify directly. The presence of multidrug-resistant bacteria is the primary reason that patients with ventilator-associated pneumonia receive inappropriate antimicrobial therapy. Empirical antimicrobial therapy for ventilator-associated pneumonia should be initiated promptly and should have a broad spectrum that covers all potential antimicrobial-resistant pathogens. Delaying the start of therapy or modifying an inappropriate antimicrobial regimen does not improve outcome, probably because the change comes too late to redirect the course of illness. Timely empirical therapy with highly effective agents that are rapidly bactericidal could minimize the emergence of resistance. Broad spectrum therapy should be streamlined (i.e., de-escalated), as appropriate, on the basis of microbiological data and clinical response. Switching to narrower spectrum therapy that is directed by culture results may minimize the emergence of resistance. For some patients, clinical response will allow a shortening of the duration of antimicrobial therapy. PMID- 16355321 TI - The relationship between antimicrobial resistance and patient outcomes: mortality, length of hospital stay, and health care costs. AB - There is an association between the development of antimicrobial resistance in Staphylococcus aureus, enterococci, and gram-negative bacilli and increases in mortality, morbidity, length of hospitalization, and cost of health care. For many patients, inadequate or delayed therapy and severe underlying disease are primarily responsible for the adverse outcomes of infections caused by antimicrobial-resistant organisms. Patients with infections due to antimicrobial resistant organisms have higher costs (approximately 6,000-30,000 dollars) than do patients with infections due to antimicrobial-susceptible organisms; the difference in cost is even greater when patients infected with antimicrobial resistant organisms are compared with patients without infection. Strategies to prevent nosocomial emergence and spread of antimicrobial-resistant organisms are essential. PMID- 16355322 TI - The role of antimicrobial management programs in optimizing antibiotic prescribing within hospitals. AB - Managing serious infections is a balance between providing timely and appropriate broad-spectrum empirical therapy for individual patients, which has been consistently shown to improve outcomes, and reducing unnecessary use of antimicrobial agents, which may contribute to the development of antimicrobial resistance. To control the spread of antimicrobial resistance, hospitals commonly implement programs designed to optimize antimicrobial use, supported by infection control measures. Hospital-based antimicrobial management programs--also called "antimicrobial stewardship programs"--are primarily based on education coupled with a "front-end" approach (i.e., restricting the availability of selected antimicrobial agents) or a "back-end" approach (i.e., reviewing broad-spectrum empirical therapy and then streamlining or discontinuing therapy, as indicated, on the basis of culture and susceptibility testing results and clinical response). Institutional efforts to optimize antimicrobial use should concentrate on patient outcomes, should have multidisciplinary support, and should use a combination of interventions customized to the needs, resources, and information technology infrastructure of the health care institution. PMID- 16355323 TI - Efficacy of machine laundering to eradicate head lice: recommendations to decontaminate washable clothes, linens, and fomites. AB - The efficacy of machine laundering to eradicate head lice should be determined. Viable lice and nits were machine laundered using 3 washing programs (with water temperatures of 40 degrees C, 50 degrees C, and 60 degrees C), with and without detergent, and the results were compared with results for control lice and nits. A drying program was also used. Either washing done with a water temperature of at least 50 degrees C or drying is necessary to kill head lice and nits. PMID- 16355324 TI - A large, multiple-restaurant outbreak of infection with Shigella flexneri serotype 2a traced to tomatoes. AB - BACKGROUND: Foodborne outbreaks of Shigella infection are uncommon and tomatoes are an unusual vehicle. We describe a large, multiple-restaurant outbreak of Shigella flexneri serotype 2a infection that was associated with tomatoes. METHODS: We conducted nationwide surveillance and a case-control study, collected fecal specimens for culture, and measured the survival of the outbreak strain of S. flexneri in tomatoes. RESULTS: We interviewed 306 of 886 ill restaurant patrons and 167 control subjects. Matched univariate analysis showed that several food items were associated with illness, but only tomatoes remained significant in multivariate models. Illness peaked at each restaurant within 24 h after the arrival of hand-sorted bruised and overripe tomatoes from a new distributor; all patient isolates that were tested were indistinguishable by PFGE. Sliced tomatoes from the distributor were inoculated with the outbreak strain, and viable S. flexneri were recovered for 72 h. CONCLUSION: To prevent such outbreaks, persons with shigellosis should be excluded from handling food at all points along the distribution chain. PMID- 16355325 TI - Control and outcome of a large outbreak of colonization and infection with glycopeptide-intermediate Staphylococcus aureus in an intensive care unit. AB - BACKGROUND: Glycopeptide-intermediate Staphylococcus aureus (GISA) is emerging as a cause of nosocomial infection and outbreaks of infection and colonization in intensive care units (ICUs). We describe an outbreak of GISA colonization/infection and the ensuing control measures in an ICU and investigate outcomes of the affected patients. METHODS: We describe an outbreak of GISA colonization and infection that affected 21 patients in a medical ICU at a tertiary care teaching hospital, as well as the measures taken to eradicate the GISA strain. RESULT: Recognition of the outbreak was difficult. Infections, all of which were severe, were diagnosed in 11 of 21 patients. Patient isolation and barrier precautions failed when used alone. Addition of a stringent policy of restricted admissions, twice daily environmental cleaning, and implementation of hand decontamination with a hydroalcoholic solution led to outbreak termination. This was associated with increases in workload, despite a marked decrease in the number of admissions. CONCLUSION: This first description of a large outbreak of GISA colonization and infection underlines the importance of routine GISA-strain detection when methicillin-resistant S. aureus is isolated. Outbreak control may be difficult to achieve. PMID- 16355326 TI - Staphylococcus aureus: times, they are a-changin'. PMID- 16355327 TI - A clinical and microbiological comparison of Staphylococcus aureus toxic shock and scalded skin syndromes in children. AB - BACKGROUND: This study was designed to compare the clinical characteristics, toxin expression, virulence factors, and antimicrobial susceptibilities of staphylococci isolated from Taiwanese children with staphylococcal toxic shock syndrome (STSS) and staphylococcal scalded skin syndrome (SSSS). METHODS: Demographic characteristics, hospital course, and outcomes of the children were analyzed. Toxin-specific and virulence genes of the staphylococci were detected by polymerase chain reaction amplification. Antimicrobial susceptibilities were determined by disk diffusion and the Etest. RESULTS: Staphylococcus aureus was isolated from 16 children (6 in the STSS group and 10 in the SSSS group). Children with STSS tended to be older than those with SSSS, had a longer duration of hospitalization, and a much higher mortality rate. Community-associated methicillin-resistant S. aureus was isolated from 11 (68.8%) of 16 children. All of these isolates contained the ermB and mecA genes, but none had the mefA gene. All 16 isolates tested positive for the fnbA gene. The pvl and seb genes were more frequently found among S. aureus from the STSS group, compared with S. aureus from the SSSS group. We found that 67% (4 of 6) of the STSS isolates were genetically related. All of the S. aureus isolates were susceptible to vancomycin, gentamicin, doxycycline, and trimethoprim-sulfamethoxazole. Most isolates were resistant to clindamycin (63%), oxacillin (69%), and clarithromycin (81%). CONCLUSIONS: The most distinguishing feature of these isolates is the greater frequency of pvl and seb carriage among those from the STSS group. Most of the isolates were community-associated methicillin-resistant S. aureus that were highly resistant to macrolides but susceptible to trimethoprim sulfamethoxazole. Vancomycin remains the initial drug of choice for treatment of STSS and SSSS. More studies are needed to determine the efficacy of trimethoprim sulfamethoxazole in children with these syndromes. PMID- 16355328 TI - Diagnostic and clinical implications of anorectal lymphogranuloma venereum in men who have sex with men: a retrospective case-control study. AB - BACKGROUND: Recently, outbreaks of anorectal lymphogranuloma venereum (LGV) have occurred among men who have sex with men (MSM). This study identifies risk factors and clinical predictors of LGV to determine the implications for clinical practice. METHODS: The Chlamydia trachomatis serovars for all MSM who had anorectal chlamydia diagnosed at a sexually transmitted infection clinic in Amsterdam, The Netherlands, in 2002 and 2003 were retrospectively typed; 87 persons were infected with C. trachomatis serovar L2b and received a diagnosis of LGV. MSM infected with C. trachomatis serovars A-K and who thus had non-LGV anorectal chlamydia (n = 377) and MSM who reported having receptive anorectal intercourse but who did not have anorectal chlamydia (n = 2677) served as 2 separate control groups. Risk factors and clinical predictors were analyzed by multivariate logistic regression. Receiver operating characteristic curves were used to determine clinical relevance. RESULTS: HIV seropositivity was the strongest risk factor for LGV (odds ratio for patients with LGV vs. those with non-LGV chlamydia, 5.7 [95% confidence interval, 2.6-12.8]; odds ratio for patients with LGV vs. control subjects without chlamydia, 9.3 [95% confidence interval, 4.4-20.0]). Proctoscopic findings and elevated white blood cell counts in anorectal smear specimens were the only clinically relevant predictors for LGV infection (area under the curve of the receiver operating characteristic curve, > 0.71). Use of these 2 parameters and HIV infection status provided the highest diagnostic accuracy (for MSM with anorectal chlamydia, the area under the curve was > 0.82; sensitivity and specificity were 89% and 50%, respectively). CONCLUSIONS: LGV testing is recommended for MSM with anorectal chlamydia. If routine LGV serovar typing is unavailable, we propose administration of syndromic LGV treatment for MSM with anorectal chlamydia and either proctitis detected by proctoscopic examination, > 10 white blood cells/high-power field detected on an anorectal smear specimen, or HIV seropositivity. PMID- 16355329 TI - Vancomycin-resistant enterococcal colonization and infection in liver transplant candidates and recipients: a prospective surveillance study. AB - BACKGROUND: Vancomycin-resistant enterococcal (VRE) infections cause significant morbidity and mortality among patients undergoing liver transplantation. We performed a prospective study among patients awaiting transplantation to assess rates, risk factors, and outcomes associated with VRE colonization before and after transplantation. METHODS: All adults on the transplantation waiting list from 2000-2003 were eligible. Demographic, historical, and laboratory data, as well as stool samples to be analyzed for VRE, were collected at enrollment and every 4-6 months thereafter until transplantation. After transplantation, samples were obtained every 3 days during hospitalization and were analyzed for VRE; outcomes were assessed at 90 days. RESULTS: Overall, 375 patients were enrolled in our study, and 142 received transplants. VRE colonization occurred in 50 (13%) of 375 patients before transplantation and was independently associated with treatment with antianaerobic antimicrobials, third-generation cephalosporins, proton pump inhibitors, or neomycin; having a recent endoscopic retrograde cholangiopancreatogram or paracentesis procedure; and admission to the liver unit. Of these 50 patients, 22 (44%) received a transplant, and 7 (32%) of 22 developed a VRE infection after transplantation. An additional 22 patients (18%) who were not colonized before transplantation acquired VRE after transplantation; VRE infection developed in 5 (23%) of these patients. Patients colonized with VRE either before or after transplantation had longer stays in the intensive care unit and the hospital. Mortality at 90 days was significantly greater among those who acquired VRE after transplantation (5 [23%] of 22), compared with those who had VRE colonization before transplantation (2 [9%] of 22). CONCLUSIONS: Liver transplantation candidates with VRE colonization before transplantation experience greater morbidity but not greater mortality, compared with noncolonized candidates. Transplant recipients who acquire VRE after transplantation have a higher mortality rate than noncolonized recipients. Strategies should be implemented to reduce nosocomial VRE acquisition after transplantation among this vulnerable group. PMID- 16355330 TI - Human parechovirus infections in Dutch children and the association between serotype and disease severity. AB - BACKGROUND: Human parechoviruses (HPeVs) are members of the family Picornaviridae and are classified into 3 known serotypes: HPeV1, HPeV2, and the recently identified HPeV3. HPeV1 and HPeV2 infections are most commonly associated with mild respiratory or gastrointestinal symptoms and occasionally with severe disease conditions, such as flaccid paralysis and encephalitis. HPeV3 infection has been associated with transient paralysis and neonatal infection and has until now only been reported in Japan and Canada. METHODS: Culture isolates considered to be enterovirus on the basis of cell culture but that were found to be enterovirus negative by 5' untranslated region reverse-transcriptase polymerase chain reaction (5'UTR RT-PCR) during the period December 2000 through January 2005 were selected. Isolates were tested by HPeV 5'UTR RT-PCR and were genotyped by sequencing the VP1 region. Phylogenetic analysis was performed, and the association with clinical symptoms was established. RESULTS: Thirty-seven (12%) of the 303 isolates that tested positive for enterovirus by cell culture were in fact HPeV. The majority of the HPeV-positive isolates (n = 27) could be identified as HPeV1. The remaining 10 isolates, which were grown from samples obtained in 2001, 2002, and 2004, could be typed as the recently identified HPeV3. HPeV was exclusively detected in children aged < 3 years. Children infected with HPeV3 were significantly younger than children infected with HPeV1, and sepsis-like illness and central nervous system involvement were more frequently reported in children infected with HPeV3. CONCLUSIONS: We report HPeV infections in young children during the period of 2000-2005 and show an association between HPeV3 infection and sepsis-like illness and central nervous system involvement in neonates. PMID- 16355331 TI - The association of respiratory viruses, temperature, and other climatic parameters with the incidence of invasive pneumococcal disease in Sydney, Australia. AB - BACKGROUND: Increases in incidence of invasive pneumococcal disease (IPD) during the colder months of the year in temperate regions are well recognized, but few detailed studies of possible interactions are available. We examined the relationship between virus activity, climatic parameters, and IPD during a winter in which there were separate peak incidences of influenza and respiratory syncytial virus (RSV) infection. METHODS: We performed an ecological study that correlated population-based data on IPD and respiratory virus activity in the year 2000 in metropolitan New South Wales, Australia, with climatic parameters, including weekly mean maximum and minimum temperature, relative humidity, rainfall, and wind speed. RESULTS: In children, RSV activity was significantly positively correlated with IPD activity (r = 0.578; P = .002) but not with influenza virus activity. There was a weak inverse relationship between parainfluenza virus activity and IPD activity (r = -0.401; P = .043) and a stronger inverse relationship between weekly mean maximum temperature (r = 0.458; P = .001), weekly mean minimum temperature (r = -0.437; P = .001), and IPD activity. In adults, there was no significant correlation between RSV or influenza virus activity alone and IPD, but the combination of RSV and influenza was significantly correlated with IPD (r = 0.481; P = .013). CONCLUSIONS: This study suggests that RSV infection and influenza contribute to IPD incidence peaks differently for children than for adults. Data from other geographic areas and more rigorous study designs are required to confirm these findings. PMID- 16355332 TI - Outcome of prosthetic joint infection in patients with rheumatoid arthritis: the impact of medical and surgical therapy in 200 episodes. AB - BACKGROUND: Prosthetic joint infection in patients with rheumatoid arthritis is a serious complication of total joint arthroplasty. Little information is available on the outcome of medical and surgical treatments of prosthetic joint infection in patients with rheumatoid arthritis. METHODS: We conducted a retrospective analysis of all patients with rheumatoid arthritis and a total hip or total knee arthroplasty infection evaluated at Mayo Clinic (Rochester, MN) between 1 January 1969 and 31 December 1995. RESULTS: A total of 200 first episodes of prosthetic joint infection in 160 patients with rheumatoid arthritis were diagnosed during the study period. Thirty-seven percent of prosthetic joint infection episodes were due to Staphylococcus aureus. Of these episodes, 23% and 19% were treated with debridement and retention of components and 2-stage exchange, respectively. The type of surgical procedure was the only analyzed clinical variable that was associated with treatment failure (P < .001). Rates of 5-year survival free of treatment failure for patients with prosthetic joint infection episodes treated with debridement and retention of components, 2-stage exchange, and resection arthroplasty were 32% (95% confidence interval [CI], 21%-49%), 79% (95% CI, 66% 93%), and 61% (95% CI, 49%-74%), respectively. CONCLUSIONS: S. aureus is the most common pathogen among patients with rheumatoid arthritis with prosthetic joint infection. Two-stage exchange was used in only 19% of the prosthetic joint infection episodes among patients with rheumatoid arthritis during the study period, but it was associated with the best outcome. The variable most strongly associated with the outcome was the type of surgical procedure. PMID- 16355333 TI - Penicillins for treatment of pneumococcal pneumonia: does in vitro resistance really matter? AB - BACKGROUND: The rate of in vitro bacterial resistance to antimicrobial agents is escalating among pathogens that cause the most serious respiratory tract infections. Many reports published during the past few years suggest that this has direct clinical implications. In particular, resistance of Streptococcus pneumoniae to beta-lactam antibiotic therapy has assumed a prominent role in the evolution of guidelines for the initial treatment of respiratory tract infection. METHODS: I conducted a critical review of the published medical literature. RESULTS: There is only a single report of documented microbiologic failure of parenteral penicillin-class antibiotics in the treatment of pneumococcal pneumonia in patients with or without bacteremia, whereas there are numerous well documented reports of treatment failure with quinolone-class (n > or = 21) and macrolide-class (n > or = 33) antibiotics in the treatment of pneumococcal pneumonia. CONCLUSION: The recommended optimal in-hospital therapy for community acquired pneumonia should continue to be a beta-lactam antibiotic (penicillin, aminopenicillin, cefotaxime, or ceftriaxone) administered with a macrolide or a fluoroquinolone agent for adjunctive treatment of infection with potential atypical pathogens. PMID- 16355334 TI - Does penicillin remain the drug of choice for pneumococcal pneumonia in view of emerging in vitro resistance? PMID- 16355335 TI - Dihydropteroate synthase mutations in the folP1 gene predict dapsone resistance in relapsed cases of leprosy. AB - Molecular detection was compared with the mouse footpad inoculation test for detection of dapsone resistance in 38 strains of Mycobacterium leprae. Mutations of the folP1 gene (at codons 53 or 55) were found in 6 of 6 strains with high level resistance, in 3 of 4 strains with intermediate-level resistance, and in 1 of 6 strains with low-level resistance, but not in 22 dapsone-susceptible strains. In cases of infection with strains of M. leprae carrying the folP1 mutation, therapy with dapsone may be replaced by therapy with a fluoroquinolone. PMID- 16355336 TI - Current treatment strategies for disseminated candidiasis. AB - The incidence of disseminated candidiasis has increased dramatically over the past several decades. Fortunately, in recent years, a variety of new antifungal agents have become available to treat these infections. On the basis of efficacy, safety, and cost considerations, fluconazole is the agent of choice for the empirical treatment of disseminated candidiasis in nonneutropenic, hemodynamically stable patients, unless a patient is suspected to be infected with an azole-resistant species (i.e., Candida glabrata or Candida krusei). For hemodynamically unstable or neutropenic patients, agents with broader species coverage, such as polyenes, echinocandins, or, possibly, voriconazole, are preferred for empirical treatment of candidemia. Modification of the initial, empirical regimen depends on the response to therapy and the subsequent identification of the species of the offending pathogen. Echinocandins or high dose polyenes are preferred for the treatment of infections with C. glabrata or C. krusei. Central venous catheters should be removed from all patients who have disseminated candidiasis, if feasible, and antifungal therapy should be administered to all patients who have candidemia or proven candidiasis. PMID- 16355337 TI - Response to antiretroviral therapy in HIV-infected patients attending a public, urban clinic in Kampala, Uganda. AB - BACKGROUND: Access to antiretroviral therapy and human immunodeficiency virus (HIV) care is increasing in resource-limited settings. We evaluated clinical, behavioral, and demographic risk factors associated with virologic suppression in a public, urban clinic in Kampala, Uganda. METHODS: We conducted a cross sectional, observational study of 137 HIV-infected patients who were receiving antiretroviral therapy at the infectious diseases clinic at Mulago Hospital (Kampala). We measured the prevalence of viral suppression, evaluated risk factors associated with virologic failure, and documented phenotypic resistance patterns and genotypic mutations. RESULTS: A total of 91 (66%) of 137 participants had an undetectable viral load (< 400 copies/mL) after a median duration of 38 weeks (interquartile range, 24-62 weeks) of antiretroviral therapy. Median CD4 cell count was 163 cells/mm3 (interquartile range, 95-260 cells/mm3). The majority of the patients (91%) were treated with nonnucleoside reverse-transcriptase inhibitor-based 3-drug regimens. In multivariate analysis, treatment with the first antiretroviral regimen was associated with viral suppression (odds ratio, 2.6; 95% confidence interval, 1.1-6.1). In contrast, a history of unplanned treatment interruption was associated with virologic treatment failure (odds ratio, 0.2; 95% confidence interval, 0.1-0.6). Of 124 participants treated with nonnucleoside reverse-transcriptase inhibitors, 27 (22%) were documented to have experienced virologic treatment failure. The most common mutation detected was K103N (found in 14 of 27 patients with virologic treatment failure). CONCLUSIONS: Although many HIV-infected people treated in Kampala, Uganda, have advanced HIV disease, the majority of patients who received antiretroviral therapy experienced viral suppression and clinical benefit. Because of the frequent use of nonnucleoside reverse-transcriptase inhibitor based therapy, the majority of resistance was against this drug class. In resource-limited settings, initiation of therapy with a potent, durable regimen, accompanied by stable drug supplies, will optimize the likelihood of viral suppression. PMID- 16355338 TI - Universal access to antiretroviral therapy: when, not if. PMID- 16355339 TI - Do type and duration of antiretroviral therapy attenuate liver fibrosis in HIV hepatitis C virus-coinfected patients? AB - BACKGROUND: This study aimed to determine whether type and duration of therapy for human immunodeficiency virus (HIV) infection attenuates liver fibrosis in patients with HIV and hepatitis C virus (HCV) coinfection. METHODS: Patients with HCV monoinfection (group 1) and HIV-HCV coinfection were retrospectively selected; the latter patients were classified into the following 3 groups: group 2, patients who received no therapy or only nucleoside reverse-transcriptase inhibitors (NRTIs); group 3, those who received highly active antiretroviral therapy (HAART); and group 4, those who initially received NRTIs followed by HAART. Fibrosis stage (scale, 0-6) and necroinflammatory score (scale, 0-18) were assessed according to the Ishak system. Data are presented as mean +/- standard deviation. RESULTS: Three hundred eighty-one patients (296 HCV-monoinfected patients and 85 HIV-HCV-coinfected patients) were recruited. The durations of HIV therapy before liver biopsy was performed for groups 2, 3, and 4 were 3.8 +/- 2.8, 3.3 +/- 1.8, and 6.6 +/- 2.2 years. The time from HIV diagnosis to HAART initiation was shorter for group 3 than for group 4 (9.1 +/- 7.3 vs. 34.1 +/- 13.1 months; P < .0001). Groups 1 and 3 had similar fibrosis stages (3.1 +/- 2 vs. 3.4 +/- 2.4), rates of fibrosis progression (0.13 +/- 0.09 vs. 0.16 +/- 0.11 per year), and necroinflammatory scores (6.1 +/- 1.8 vs. 6.1 +/- 2.0). Groups 2 and 4 had significantly more-advanced liver disease, as determined by fibrosis stage (4.6 +/- 1.8 vs. 4.3 +/- 2.0; P < .0009), rate of fibrosis progression (0.24 +/- 0.11 vs. 0.20 +/- 0.10 per year; P < .0001), and prevalence of cirrhosis (68% vs. 55%; P < .006), compared with group 1. CONCLUSIONS: HIC-HCV coinfected subjects who receive HAART as their sole form of therapy have liver histology findings comparable to those for HCV-monoinfected patients. A similar degree of benefit is not observed for HIV-HCV-coinfected patients who receive no therapy, NRTIs, or HAART after NRTIs, despite having a longer duration of therapy. PMID- 16355340 TI - Liver fibrosis and antiretroviral therapy. PMID- 16355341 TI - Body fat and other metabolic effects of atazanavir and efavirenz, each administered in combination with zidovudine plus lamivudine, in antiretroviral naive HIV-infected patients. AB - BACKGROUND: Protease inhibitor treatment of human immunodeficiency virus (HIV) infected individuals has been linked to the development of lipodystrophy. The effects of atazanavir on body fat distribution and related metabolic parameters were examined in antiretroviral-naive patients. METHODS: HIV-positive patients with CD4 cell counts > or = 100 cells/mm3 were randomized to 1 of 2 treatment arms: (1) atazanavir, 400 mg given once daily, plus efavirenz placebo; or (2) efavirenz, 600 mg given once daily, plus atazanavir placebo; each drug was administered with fixed-dose zidovudine (300 mg) and lamivudine (150 mg) given twice daily, and patients were treated for at least 48 weeks. Fat distribution measurements (visceral adipose tissue [VAT], subcutaneous adipose tissue [SAT], and total adipose tissue [TAT], as measured by computed tomography; and appendicular fat, truncal fat, and total fat levels, as measured by dual-energy x ray absorptiometry), metabolic measurements (cholesterol and fasting triglyceride levels), and measurements of insulin resistance (fasting glucose and fasting insulin levels) were made at baseline and at week 48 of treatment for a subgroup of 111 atazanavir recipients and 100 efavirenz recipients. RESULTS: Atazanavir and efavirenz treatments resulted in minimal to modest increases in fat accumulation, as measured by VAT, SAT, TAT, appendicular fat, truncal fat, and total fat levels; results were comparable in both arms. In addition, atazanavir was associated with none of the metabolic abnormalities seen with many other protease inhibitors. CONCLUSIONS: Use of atazanavir for 48 weeks neither resulted in abnormal fat redistribution in antiretroviral-naive patients nor induced other metabolic disturbances commonly associated with HIV-related lipodystrophy. Longer term assessments (e.g., at 96 weeks) will be important to confirm these findings. PMID- 16355342 TI - HIV-associated lipoatrophy: what are the kinder, gentler agents? PMID- 16355343 TI - Tenofovir-associated acute and chronic kidney disease: a case of multiple drug interactions. AB - Tenofovir therapy in patients with human immunodeficiency virus (HIV) infection has been associated with acute renal failure (ARF) and Fanconi syndrome. In the past 2 years, we diagnosed tenofovir-associated ARF in 5 HIV-infected patients who were receiving tenofovir therapy and who had classic findings of acute tubular necrosis, and we compared findings for our patients with data on 22 patients described in the literature. The mean serum creatinine level increased from 0.9 to 3.9 mg/dL, and it decreased to 1.2 mg/dL during recovery. ARF resolved in 22 of 27 patients after discontinuation of tenofovir therapy. The most common drugs given with tenofovir were ritonavir or lopinavir-ritonavir (21 of 27 patients), atazanavir (5 of 27 patients), and didanosine (9 of 27 patients). Tenofovir-associated ARF manifests as acute tubular necrosis that may not resolve with tenofovir withdrawal. Tenofovir is associated with multiple drug interactions, leading to an increased risk of ARF. Frequent monitoring of renal function is warranted for any patient receiving these combinations. PMID- 16355344 TI - Plasma levels of atazanavir and the risk of hyperbilirubinemia are predicted by the 3435C-->T polymorphism at the multidrug resistance gene 1. AB - The 3435C-->T polymorphism at the multidrug resistance gene 1 (MDR1) was examined in 74 patients with human immunodeficiency virus who initiated atazanavir therapy. The MDR1 genotype distribution at position 3435 was 28% CC, 45% CT, and 27% TT. Plasma levels of atazanavir were significantly higher in patients with genotype CC than in those with CT or TT, and bilirubin levels correlated with atazanavir concentrations. PMID- 16355345 TI - Coinfection with HIV and hepatitis C virus and immune restoration during HAART. PMID- 16355346 TI - "Strange things I have in head": evidence of prion disease in Shakespeare's Macbeth. PMID- 16355347 TI - Acute cholecystitis complicating mumps. PMID- 16355348 TI - Symptomatic Schistosoma mansoni infection as an immune restoration phenomenon in a patient receiving antiretroviral therapy. PMID- 16355349 TI - Revisiting combination antibiotic therapy for community-acquired invasive Streptococcus pneumoniae pneumonia. PMID- 16355350 TI - Rash decisions about southern tick-associated rash illness and Lyme disease. PMID- 16355352 TI - Self-screening for rectal sexually transmitted infections: human papillomavirus. PMID- 16355353 TI - [Topiramate or flunarizine in the preventive treatment of migraine. A comparative study of two series of cases]. AB - INTRODUCTION: Both flunarizine and, more recently, topiramate have proved to be effective in the prophylaxis of migraine. AIM: To compare two independent groups of patients with similar clinical and demographic characteristics who were treated with flunarizine or topiramate as the preferred drug in the prevention of their migraines and to evaluate effectiveness and safety according to the medication they received. PATIENTS AND METHODS: All the patients reported more than four episodes a month and/or transformed migraine according to Silberstein's criteria, and had never received prophylactic treatment. Data determined: the number and average number of migraines in the previous month and at the fourth month of treatment, and the rate of respondents. RESULTS: A total of 125 patients were included in each group. No significant differences were found between the groups as regards mean age or the average number of migraines in the previous month. With both drugs there was a significant decrease (0.0001) in the mean number of episodes in the fourth month of treatment, but with no significant difference between them: topiramate (5.88 +/- 3.7 to 2.1 +/- 2.5) and flunarizine (5.24 +/- 3.2 to 2.3 +/- 2.7). The mean reduction in the number of migraines at the fourth month was 58.2 +/- 38.2% with topiramate, and 55.4 +/- 37.5% with flunarizine. The respondent rate was 71% with topiramate and 67% with flunarizine. The percentage of dropouts with topiramate (28%) was higher than with flunarizine (11%) (0.0013). With topiramate 69 patients reported side effects and 53 patients reported them with flunarizine (0.0427). CONCLUSIONS: Both drugs showed a high degree of effectiveness when used as the preferred drug in the preventive treatment of migraine. Topiramate offered better results as far as effectiveness is concerned, but also more side effects, none of which were serious. PMID- 16355354 TI - [Impact of experience on improving the surgical outcome in temporal lobe epilepsy]. AB - INTRODUCTION: Recently, we have published the results of a first surgical series of patients with temporal lobe epilepsy (TLE). We describe a posterior series of patients intervened of TLE, we compare the functional results with the previous series and we finally analyze the causes of changes. PATIENTS AND METHODS: We studied the first 22 consecutive patients surgically intervened of TLE with a minimum post-surgery follow-up of 2 years. Patients showing I and II Engel's grade were used as gold standard for evaluation of pre-surgical complementary studies. RESULTS: We have obtained better functional results: 91% patients showing Engel's grade I, 9% showing grade II and neither III nor IV grades were obtained. Pre-surgical studies changed in comparison with the previous report. The most improving change was observed in video-EEG with foramen-ovale electrodes (FOE) (37%), scalp EEG (26.6%), interictal SPECT (11.7%) and MRI (11.7%). Video EEG with FOE was the study than showed greater concordance with epileptic focus (95.5%), followed by EEG (86.4%). In 35% of cases, MRI was normal or without valid data for correct localization of focus. CONCLUSIONS: Video-EEG with FOE and TLE surgery are safety methods, which results improve with the experience. Normal or not informative MRI do not should a priori reject those patients with drug resistant TLE from surgery. PMID- 16355355 TI - [Validity of the Spanish version of the Addenbrooke's Cognitive Examination for the diagnosis of dementia and to differentiate Alzheimer's disease and frontotemporal dementia]. AB - INTRODUCTION: The Addenbrooke's Cognitive Examination (ACE) is a brief bedside test battery to detect mild dementia and differentiate frontotemporal dementia (FTD) from Alzheimer's disease (AD). AIM: To validate the ACE in Spanish. PATIENTS AND METHODS: The study evaluated the Spanish version of ACE on 128 subjects consisting in two groups a patient group (n = 76) and a control subjects group (n = 52). The patient group was divided in AD (n = 54) based on the NINCDS ADRDA criteria and FTD (n = 22) based on the Lund y Manchester criteria. All patients underwent clinical, neuropsychological, radiologic (MRI, CT, and SPECT), and laboratory evaluations. Group's differences were evaluated using ANOVA. The internal consistency of the Spanish version of the ACE was measured using the Cronbach's alpha coefficient. The discriminative capability of the Spanish version of the ACE was examined by the receiver operating characteristic (ROC) analysis. RESULTS: The cut-off score of 86 showed a sensitivity of 92% (CI 95% = 83.6-97.0) and a specificity of 96.2% (CI 95% = 86.8-99.4). The ROC curve showed higher sensitivity and specificity of the ACE than the Mini-Mental State Examination in discriminating the dementia and control group. The VLOM ratio (verbal fluency + language)/(orientation + memory) of < 1.82 discriminated for FTD and > 4.87 discriminated for AD. CONCLUSION: The Spanish version of ACE is a brief and reliable instrument for early detection of dementia in highly educated people and offers a simple objective index to differentiate AD and FTD. More studies in less educated people are warranted. PMID- 16355356 TI - [Cerebral aneurysm related to pediatric AIDS. Case report]. AB - INTRODUCTION: Aneurysms, described in pediatric AIDS, are related with bad prognosis with high mortality in 5 months. The twenty reports found in literature were considered late (mean age 9.97 years old) and associated with severe immunological involvement. The mean CD-4 count was 79.17 cells and mean viral load 222,662 copies/mL. Some authors consider this complication associated to the human immunodeficiency virus (HIV), to an opportunistic agent (citomegalovirus and Mycobacterium avium) or due to inflammatory mechanisms related with the immunological improvement. CASE REPORT: This paper present a 9 years old girl with severe AIDS (C-3) and with transient amaurosis related to transient ischemic attacks on ophthalmic artery during her immunological improvement on viral load and CD-4 counts. Necropsy showed aneurysm due to atypical micobateriosis. CONCLUSION: This report highlights the occurrence of this rare but severe HIV complication witch was associated with an opportunistic etiology during an immunological recovery, when inflammatory mediators could play a role. PMID- 16355357 TI - [Cerebral cavernomas. A review and update of aetiological, clinical and therapeutic features]. AB - INTRODUCTION: Cavernous angiomas are uncommon lesions, with a reported incidence of 0.4 to 0.8%, presenting a controversial management especially regarding their surgical treatment. AIM: To update cavernous angiomas characteristics and management through a deep review of the literature concerning their aetiology, epidemiology, history, signs and symptoms, diagnosis, and surgical and radiosurgical treatment. DEVELOPMENT AND CONCLUSIONS: Most important advances found in the recent literature include the identification of the genetic basis responsible for the familial form of cavernomatosis (CCM1, CCM2 and CCM3), the identification of the dynamic pattern of these lesions based on their pathology and imaging features, the deeper knowledge on their natural history depending on their supra/infratentorial location, and the main indications for surgical treatment and radiosurgical therapy suggested by the recent series. PMID- 16355358 TI - [The neurophysiology of the interaction between attention and episodic memory: a review of studies into the visual mode]. AB - INTRODUCTION: Attention is the process that provides us with the capacity to select perceptions or internal representations for conscious processing and action. Given that episodic memory is where our personal experiences are stored together with the context they take place in, it has been observed that disturbances affecting attention, whether they are due to neurological disorders or experimental manipulation, deteriorate the storage of episodic information. AIMS: The aim of this study is to explain, from a neurophysiological perspective, how the processes of attention and memory interact with each other and which areas of the brain are involved in those processes. This objective is motivated by the fact that, despite empirical evidence supporting the interaction between the two processes, to our knowledge no reviews dealing with such an interaction have been published to date in the literature. DEVELOPMENT: In this work attention and memory are defined in terms of the neurophysiological changes that take place when the two processes occur. In what follows we offer a description of the systems that constitute attention, as well as those involved in the acquisition and recognition of information in episodic memory. Finally, we propose a possible explanation of how the mechanisms of interaction between these two processes work. CONCLUSIONS: The system of attention and the system of episodic memory are brought into play at the same time; the prefrontal cortex, in particular, plays a role in both processes. Attention seems to precede the mechanisms behind the acquisition of information while it is held during recognition. PMID- 16355359 TI - [Ischemic stroke in childhood and adolescence. A study of 46 cases in the south of Brazil]. AB - INTRODUCTION: Ischaemic stroke (IS) in childhood and adolescence is a condition that has received little attention from researchers and presents a multifactorial aetiology. In recent decades the rates of incidence seem to have risen, but underdiagnosis is still commonplace. It may appear as the first symptom of systemic disease in childhood, with the resulting devastating effects. Prothrombotic disorders represent important causal factors of ischaemic attacks in childhood. AIMS: The aims of this study were to identify the factors associated to IS in infancy and adolescence, as well as to establish the causation and to determine the abnormalities in the coagulation proteins of the patients that were studied. PATIENTS AND METHODS: A total of 46 patients diagnosed with IS, between 0 and 18 years of age, were included in the study over the period between March 2002 and September 2003. A number of lab tests were carried out, including analysis of coagulation proteins and echocardiograms. RESULTS: Neonatal IS was found in almost a third of the cases and practically half the sample had some disease prior to the ischaemic event. In all, 20% of the patients showed no IS-associated disorders. The most frequent clinical presenting symptoms were convulsions and hemiparesis. Anomalies were detected in proteins S and C in 22 and 17% of the sample, respectively. The middle cerebral artery was the most affected vascular region. CONCLUSIONS: IS-associated disorders were identified in most of the children and adolescents that were studied; moreover, prothrombotic risk factors are of great significance in the research into cases. PMID- 16355360 TI - [Endovascular treatment of carotid disease. The state of the art, technical aspects and occupational training]. AB - AIMS: The aim of this study was to carry out a review of the state of the art in the endovascular treatment of carotid disease, taking into account the findings currently available, its indications, the technical aspects linked to the intervention as well as those related to the occupational training of the specialists involved in performing the technique. DEVELOPMENT: Surgical treatment of carotid artery disease can be of benefit to symptomatic patients with stenoses above 70% and to subgroups of patients with symptomatic stenoses between 50-69%. The benefit of carotid endarterectomy in asymptomatic patients is the object of a great deal of controversy that is concerned with the reduction in risk that is obtained and also the large number of patients to be treated in order to prevent the occurrence of ischaemic events. The endovascular treatment of carotid disease comes to the fore, then, as an alternative to surgical treatment. At the present time, a number of randomised multicentre studies are being conducted that will allow the two techniques to be compared in homogeneous groups of patients. Preliminary data nevertheless seem to suggest that this technique offers a number of benefits, especially in groups of subjects with a high surgical risk, and thus they could allow the indications for revascularisation to be extended. We analyse the technical and medical aspects linked to this procedure, the findings from studies carried out to date, its indications and the occupational training of the specialists involved in performing the technique. CONCLUSIONS: The endovascular treatment of carotid disease constitutes an alternative to surgical treatment in specifically selected patients. The potential increase in its indications is conditioned by the results from the multicentre studies currently being carried out. PMID- 16355361 TI - [Transdural herniation of the spinal cord: a rare cause of myelopathy]. PMID- 16355363 TI - [Variability in clinical practice in cases of headaches during infancy: the need for studies into how to adjust clinical practice to the scientific evidence]. PMID- 16355364 TI - [Treatment with valproate and hepatic steatosis]. PMID- 16355365 TI - [Nitrogen monoxide in malignant astrocytomas]. PMID- 16355366 TI - Estimating the optimal radiotherapy utilization for carcinoma of the central nervous system, thyroid carcinoma, and carcinoma of unknown primary origin from evidence-based clinical guidelines. AB - BACKGROUND: In this one in a series of articles, the objective was to estimate the ideal proportion of patients with cancer who should receive radiotherapy at least once during the course of their illness based on the best available evidence. This estimate should be useful in planning for future radiotherapy facilities. Optimal rates of radiotherapy for patients with central nervous system (CNS) carcinoma, thyroid carcinoma, or carcinoma of unknown primary site (CUP) have not been studied previously. METHODS: A systematic review of evidence based treatment guidelines for the treatment of CNS carcinoma, CUP, and thyroid carcinoma was undertaken. An optimal radiotherapy utilization tree was constructed for each of these malignancies depicting the indications for radiotherapy at various stages of disease. The proportion of patients who had clinical attributes that indicated a possible benefit from radiotherapy was calculated by adding epidemiological data to the radiotherapy utilization tree. The optimal proportion of patients who should receive radiotherapy was then calculated using specialized decision-analysis software. Sensitivity analyses using univariate analysis and Monte Carlo simulations were performed. RESULTS: The optimal rates of radiotherapy utilization for carcinoma of the CNS, thyroid carcinoma, and CUP were 92%, 10%, and 61%, respectively. Comparison with actual rates of utilization in South Australia, Sweden, and the U.S. suggested an under utilization of radiotherapy for CNS carcinoma and CUP. However, the actual rates of radiotherapy for thyroid carcinoma exceeded the optimal rate for some jurisdictions, although some data may have included radioactive iodine, which was not included in the current project. CONCLUSIONS: It was possible to estimate optimal radiotherapy utilization rates based on evidence. This methodology allowed a comparison of optimal rates with actual rates to identify areas in which improvements in the evidence-based use of radiotherapy can be made, and it may provide valuable data for future radiotherapy service planning. PMID- 16355368 TI - Following a crush and a kiss: why is it lonely out there? PMID- 16355367 TI - ACC/AHA/SCAI 2005 guideline update for percutaneous coronary intervention- summary article: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (ACC/AHA/SCAI Writing Committee to update the 2001 Guidelines for Percutaneous Coronary Intervention). PMID- 16355369 TI - Growth conelike sensorimotor structures are characteristic features of postmigratory, premyelinating oligodendrocytes. AB - During development, postmigratory, premyelinating oligodendrocytes extend processes that navigate through the central nervous system (CNS) environment, where they recognize a number of extracellular cues, including axonal segments to be myelinated. Ultimately this recognition event leads to the formation of the CNS myelin sheath. However, the morphological structures and molecular mechanisms that control such oligodendroglial pathfinding are poorly understood. Here we show that postmigratory, premyelinating oligodendrocyte processes possess at their distal tips expansions that ultrastructurally resemble growth cones of postmigratory neurons and that we will refer to as OLG-growth cones. OLG-growth cones are highly motile and capable of mediating process outgrowth, retraction, and branching. In addition, they express regulators of cytoskeletal organization, GAP43 and cofilin, that are known to mediate neuronal growth cone navigation. In a choice situation, processes of postmigratory, premyelinating oligodendrocytes and their OLG-growth cones have the ability to selectively avoid a nonpermissive substrate, that is, collagen IV. Thus, our findings provide, for the first time, a detailed characterization of sensorimotor structures present at the tips of postmigratory, premyelinating oligodendrocyte processes. Furthermore, the data presented here suggest that, although the cellular mechanisms involved in growth cone steering may be similar for postmigratory neuronal and oligodendroglial cells, extracellular cues may be interpreted in a cell-type-specific fashion. PMID- 16355370 TI - Bystander killing of neurons by cytotoxic T cells specific for a glial antigen. AB - To explore pathways to neuron loss in inflammatory diseases, transgenic mice expressing beta-galactosidase (beta-gal) in either astrocytes or photoreceptor cells, or both, were inoculated with activated, beta-gal-specific cytotoxic CD8 T lymphocytes (CTLs). beta-gal-positive astrocytes in brain were rapidly attacked, with particular damage in cerebellum. Substantial loss of cerebellar granule cells was found, even though these neurons did not express beta-gal. The small number of beta-gal-positive retinal astrocytes present in these mice was also rapidly destroyed by transferred CTLs, but without detectable consequences for retinal neurons. However, in mice with photoreceptor cell-specific beta-gal expression, near-total destruction of photoreceptor cells was produced by CTL transfer. Attack on photoreceptor cells displayed minimal inflammation, and onset was a week later than onset of astrocyte-directed disease. CTL transfer into F1 mice expressing beta-gal in both astrocytes and photoreceptor cells confirmed that pathogenesis directed against antigen expressed in glia versus neurons proceeded in two distinct, independent phases. beta-gal-positive retinal astrocytes were severely affected by 5 days post-transfer, followed by rapid resolution. Photoreceptor cells in the same retina were unaffected until 12 days post-transfer. The susceptibility of photoreceptor cells was not enhanced by the prior CTL attack on beta-gal-expressing retinal astrocytes. The results demonstrate that extensive bystander killing of neurons can occur in vivo as a result of direct CTL attack on surrounding astrocytes. Antigen-expressing retinal neurons were also efficiently killed by CTLs, but by a mechanism that was substantially delayed and dissociated from the killing of retinal astrocytes. PMID- 16355371 TI - Adult olfactory ensheathing glia promote the long-distance growth of adult retinal ganglion cell neurites in vitro. AB - In vivo, transplanted adult olfactory ensheathing glia (OEG) and adult Schwann cells (SC) can support the regrowth of at least some transected axons within adult CNS neuropil. In the present study, we developed an in vitro adult rat retinal explant model to explore the influence of primary adult SC and OEG on retinal ganglion cell (RGC) neurite regrowth in the presence of glial cells endogenous to the retina. Retinal quadrants were plated RGC-side down onto aclar hats coated with either pure collagen (type 1), collagen with OEG, collagen with SCs, or collagen coated with both OEG and SCs. Regrowing retinal neurites extended onto the pure collagen substrate, largely in association with astrocytes that migrated out from the explants (mean number of neurites: 144+/-65 SEM). The additional presence of OEG (669+/-122), but not SCs (97+/-41), supported the regrowth of significantly greater numbers of RGC neurites. Furthermore, this OEG stimulated regeneration was over significantly greater distances; >68% of neurites extended >500 microm from the explant, compared with explants plated onto SCs or collagen alone (15% and 29%, respectively). When OEG and SCs were co cultured the number of regenerating neurites was reduced (397+/-81) compared with the pure OEG treatment. Analysis of explants on pure collagen substrates fed with media conditioned by purified OEG or SC showed no increase in neurite outgrowth compared with control treatments, suggesting that the enhanced growth in the presence of OEG is a contact-mediated effect. The observed differences between the abilities of OEG and SC to support the growth of CNS-derived fibers in the presence of astrocytes support the suggestion that OEG may be better suited for direct transplantation into CNS neuropil following injury. PMID- 16355372 TI - Techniques for percutaneous closure of prosthetic paravalvular leaks. AB - Percutaneous transcatheter closure techniques are now routinely applied in the management of atrial and ventricular septal defects, patent ductus arteriosus, and other pathological cardiac and vascular communications. Recently, these same techniques have been applied to paravalvular defects. Reports are few; success variable and techniques vary widely. We review the current considerations and techniques of percutaneous transcatheter closure of paravalvular leaks. PMID- 16355373 TI - Mediastinum thymoma diagnosed by FNA and ThinPrep technique: a case report. AB - Thymoma is a primary tumor of the thymus epithelial cells. It may be asymptomatic or accompanied with atypical clinical symptoms or paraneoplastic syndromes, such as myasthenia gravis. The biological behavior of thymomas is unpredictable. The invasion of the capsule or the adjacent tissues is the major diagnostic criterion for the malignant behavior of these tumors. This is an interesting case of thymoma diagnosed by fine-needle aspiration biopsy (FNA) and ThinPrep technique, in a 54-year-old female patient with a history of gastric adenocarcinoma. Cytology of the mediastinum mass revealed a mixed population of epithelial cells and lymphocytes. The tumor was excised and the histopathological examination supported the cytological diagnosis. Thymomas commonly constitute a problem in differential diagnosis of mediastinum masses. FNA biopsy in correlation with ThinPrep technique and immunocytochemistry may play a significant role in clarifying the nature of these lesions and may contribute to the early management and choice of the optimal therapeutic manipulation. PMID- 16355374 TI - Salivary duct carcinoma metastatic to inguinal lymph node: a case report of salivary duct carcinoma with distant metastasis diagnosed by fine-needle aspiration. AB - Salivary duct carcinoma (SDC) is a high-grade malignant tumor exhibiting aggressive growth with early regional and distant metastasis. We report a case of SDC in a 53-yr-old male with distant metastasis to an inguinal lymph node. The diagnosis of the primary tumor as well as the metastatic lesion was accomplished by fine-needle aspiration (FNA). Aggressive clinical management appears to be the main therapeutic option for long-term survival. Therefore, establishing an accurate preoperative diagnosis by FNA can have both clinical and prognostic relevance. PMID- 16355375 TI - Dysbacteriosis in silver-stained cervical smears of Dutch-Moroccan immigrants: HPV infection and preneoplasia. AB - The vaginal/cervical smears of a group of Moroccan immigrants were used to compare vaginal dysbacteriosis (i.e., a bacterial population change with a decrease in lactobacilli and an increase of coccoid bacteria in vaginal/cervical smears) with Dutch women. From our archives, 779 smears from Moroccan immigrants were compared with 1,060 smears of age-matched Dutch women. For bacterial flora, Jones-Marres silver stains were used to define four groups. Koilocytosis and cervical intraepithelial neoplasia (CIN) were also recorded. The bacterial vaginal flora of Moroccan immigrants and Dutch women was different. The Moroccan women had a lower dysbacteriosis than Dutch women (3% vs. 24%). Koilocytosis and CIN were less frequent in the immigrant population. The possible synergy of a disturbed vaginal flora with human papillomavirus (HPV), HIV, or cervical preneoplasia indicates that vaginal hygiene and a normal flora may have positive effects on the uterine cervix. PMID- 16355376 TI - Effect of Bethesda 2001 on reporting of atypical squamous cells (ASC) with special emphasis on atypical squamous cells-cannot rule out high grade (ASC-H). AB - We compared the overall ASC rate and the outcomes for women with different categories of ASC before and after TBS 2001 to evaluate the impact of TBS 2001. Our laboratory reported ASC in four subcategories before TBS 2001; ASC, favor reactive (ASC-R), favor low-grade squamous intraepithelial lesion (SIL) (ASC-L), undetermined significance (ASC-US), and ASC-H. Since the implementation of TBS 2001, we have been reporting ASC as ASC-US and ASC-H. After TBS 2001, our ASC rate decreased from 6.2% to 4% (P < 0.0001). Before TBS 2001, ASC-R carried the same risk as ASC-US for underlying condyloma/CIN 1. Moreover, ASC-R was rarely associated with CIN 2 or 3. Before TBS 2001, ASC-H captured more low-grade (condyloma/CIN 1) and less high-grade (CIN 2, 3, and SCC) lesions compared to after TBS 2001. More women with ASC-H after TBS 2001 underwent colposcopy (80% vs. 71%, P < 0.05). Twenty-two percent of women with ASC-H required 2-4 colposcopies to identify the high-grade lesion. Our results confirm that consistent application of TBS 2001 terminology for ASC reduces the ASC rate, and better identifies women at high risk for CIN 2, 3, and SCC. In addition, ASC-H requires very close clinical follow-up as a significant percentage of women require more than one colposcopy to identify the high-grade lesion. PMID- 16355377 TI - Comparison of three cytologic preparation methods and immunocytochemistries to distinguish adenocarcinoma cells from reactive mesothelial cells in serous effusion. AB - We assessed whether a panel of seven antibodies is useful in the differentiation of adenocarcinoma cells (ACCs) from reactive mesothelial cells (RMCs) in effusion samples and to determine optimal specimen preparation conditions for immunocytochemical analysis of effusion samples. Immunocytochemistry (ICC) was performed on three types of effusion preparations from the same effusion specimens: ethanol-fixed smears, ethanol-fixed cell-blocks, and formalin-fixed cell-blocks. Commercially available antibodies MOC-31, Ber-EP4, CA19-9, CEA, EMA, CA125, and HBME-1 were tested on RMCs from four samples of various etiology and 15 samples of adenocarcinoma from various primary sites. Ethanol-fixed smears showed strong immunoreactivity to all antibodies tested. The immunoreactivity of ethanol-fixed and formalin-fixed cell-blocks was significantly lower with all antibodies except CA19-9. Smear preparations are more sensitive than cell-blocks for immunocytochemical study. A panel of antibodies MOC-31, Ber-EP4, CA19-9, and CEA appears to be suitable to distinguish between ACCs and RMCs. PMID- 16355378 TI - Reporting thyroid fine-needle aspiration: literature review and a proposal. AB - Reporting schemes for thyroid fine-needle aspirations in the literature were reviewed and classified according to the number of categories in the scheme and the significance of each category. The sensitivity, specificity, and positive predictive and negative values were determined for each scheme, if possible. A reporting scheme based on the probability of finding carcinoma on histology is proposed. PMID- 16355379 TI - Fine-needle aspiration of parapharyngeal lipoma. PMID- 16355380 TI - Fine-needle aspiration cytology and immunocytochemistry of orbital masses. AB - A series of 85 fine-needle aspiration (FNA) biopsies from orbital space occupying lesions of 82 patients are reviewed. A total of 32 benign lesions and 49 malignant lesions were conclusively diagnosed. In two cases the aspirates were insufficient for diagnosis. Of two cases, which were cytologically suspicious for lymphoma, a repeat FNA resulted in a conclusive diagnosis of lymphoma in one case, while the second case proved to be a pseudotumor on an open biopsy material. Of the 32 benign lesions seven were fibrosis, six pseudotumors, four epidermal cysts, four meningiomas, and three pleomorphic adenomas. The remaining cases included two hematomas, one granuloma, three inflammations, and one malformation. In 43 of 49 malignant tumors cytomorphology was corroborated with immunocytochemistry. Thirty five of these were low- or high-grade lymphomas, nine metastases, two sarcomas, two plasmacytomas, and one chloroma. All lymphomas were of B phenotype with monoclonal light chain expression. The rate of cell proliferation as measured by Ki-67 immunostaining varied between 4-25% and 30-80% for low- and high-grade lymphomas, respectively. These results confirm previous reports on the usefulness of FNA biopsy in diagnosing orbital masses and emphasize the value of immunocytochemistry in tumor characterization. PMID- 16355381 TI - Detection of Epstein-Barr virus (EBV) in the oral mucosa of renal transplant patients. AB - The purpose of this study was to determine the prevalence of Epstein-Barr virus (EBV)-DNA in the oral mucosa of renal transplant patients and observe the efficacy of mouth rinses with phosphate-buffered saline (PBS) to eliminate EBV present in the saliva. Lingual, gingival, and buccal cytobrushings were obtained from normal oral mucosa of 10 renal transplant patients and 10 normal subjects, and were examined through polymerase chain reaction (PCR), before and after rinses with PBS. EBV-DNA was detected in 86.6% of renal transplant recipients and in 46.6% of healthy subjects. No significant difference was observed between oral scrapes obtained before and after rinses with PBS with regard to detection of EBV DNA. Our results suggest that the use of PCR to detect the presence of EBV-DNA in oral mucosa in the absence of specific lesions gives rise to the problem of identifying the viral replication sites. In addition, PBS was not effective at minimizing contamination by saliva. PMID- 16355383 TI - Quantification of the cellular components of breast duct lavage samples. PMID- 16355382 TI - Vegetable cells in Papanicolaou-stained cervical smears. AB - Vegetable cells are unusual findings in Papanicolaou-stained cervical smears; these structures could be wrongly mistaken for abnormal human cells, worm eggs, or spores by a cytologist encountering the possibility of meeting those elements in cytological analysis. Five cervicovaginal smears showing similar vegetable cells have been detected over a 3-yr period (2002-2004) in the course of a population screening program for cancer of the uterine cervix in Modena (Italy) involving 32,500 women. According to the clinical histories of the patients, the vaginal pharmaceutical drugs or appliances used were of different types: vaginal lavages, pessaries, and vaginal creams. Following a careful investigation, the only substance that can lead to vegetal elements has been identified as polysaccharide galactomannan, which is one of the excipient present in the drugs used. The authors have identified the origin of these contaminants and the means of pollution, using cytological and pharmaceutical investigation. PMID- 16355384 TI - Factors influencing long-term survival following salvage total laryngectomy after initial radiotherapy or conservative surgery. AB - BACKGROUND: This retrospective study investigated survival outcomes of salvage total laryngectomy (STL) after initial radiation therapy (RT) or larynx conservation surgery (CS) at an academic center. METHODS: A chart review yielded 64 patients with STL: 53 with RT failures, six with CS failures, and five after RT + CS. Median potential follow-up after STL was 9.4 years (mean, 9.2 years; range, 0.3-17.4 years). RESULTS: Five- and 10-year actuarial overall survival (OS) after STL was 65.2% and 37.7%, respectively. Mean survival after STL was 7.2 years (median, 6.8 years; range, 0.2-17.4 years). No significant survival difference was found between the three treatment groups (p = .50). For 21 patients with nodes assessed at STL, 9-year OS was 45.4% for patients with N0 disease versus 26.7% for patients with N+ disease (p = .25). CONCLUSION: These data suggest that STL after radiation failure is associated with equivalent long term survival as STL after RT + CS or after failure of CS alone. PMID- 16355385 TI - Endoscopic CO2 laser surgery for early glottic cancer in patients who are candidates for radiotherapy: results of a prospective nonrandomized study. AB - BACKGROUND: Treatment of early glottic malignancies is controversial, particularly if postoperative endoscopy shows minimal/no residual disease. METHODS: In a prospective nonrandomized study, we performed endoscopic laser surgery with curative intent in 59 patients with early glottic carcinoma presenting for radiotherapy after diagnosis elsewhere by random biopsy or cordal stripping. We analyzed outcomes and residual cancer in the specimen. RESULTS: Clinical staging elsewhere did not correspond to (usually underestimated) the pathologic stage in 60.7%. In 22%, no tumor was identified on pathologic examination. After 35.3 months (mean), 93.2% were alive with no evidence of disease. Local control was achieved in 98.3% of the patients, and 13.5% of the patients required radiotherapy. The larynx was conserved in 98.3%. CONCLUSIONS: Biopsy/stripping are best abandoned for persistent glottic lesions. A single laser endoscopic procedure provides reliable staging and definitive treatment in most cases using fewer resources. Biopsied patients presenting for treatment should be offered laser surgery as an alternative to radiotherapy. PMID- 16355386 TI - Effects of a novel NF-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), on growth, apoptosis, gene expression, and chemosensitivity in head and neck squamous cell carcinoma cell lines. AB - BACKGROUND: Recent studies provide evidence that the constitutive activation of nuclear factor-kappa B, NF-kappaB plays a critical role in enhancing the growth of several types of malignancies, including head and neck squamous cell carcinoma (HNSCC). METHODS: In this study, we examined the effects of a newly synthesized NF-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), on growth, induction of apoptosis, gene expression, and chemosensitivity in two HNSCC cell lines (YCU H891 and KB), which expressed high levels of nuclear NF-kappaB protein. RESULTS: DHMEQ showed strong growth inhibitory effects on these two cell lines, with a 50% cell growth inhibition (IC50) concentration of approximately 20 microg/mL. These growth inhibitory effects were associated with inhibition of the NF-kappaB activity. Treatment with DHMEQ induced apoptosis in a dose-dependent manner accounting, at least in part, for the growth inhibition by DHMEQ. DHMEQ strongly inhibited cyclin D1 and vascular endothelial growth factor (VEGF) promoter activity and decreased the levels of cyclin D1 protein and VEGF mRNA in KB cells. In addition, low concentrations of DHMEQ (1.0 or 5.0 microg/mL) synergistically enhanced the cellular sensitivity of YCU-H and KB cells to cisplatin, which is a key chemotherapeutic agent in the treatment of HNSCC. CONCLUSIONS: These results suggest that DHMEQ may be effective when used alone or in combination with other agents in the treatment of HNSCC. PMID- 16355387 TI - Hyaluronan and its receptors in mucoepidermoid carcinoma. AB - BACKGROUND: Hyaluronan (HA) is a prominent extracellular matrix component undergoing continuous production and degradation. Increased HA levels have been described in a variety of tumors. The objective of this study was to examine the staining patterns of HA and two of its associated receptors (CD44 and HARE) in relation to the metastatic potential of mucoepidermoid carcinoma (MC). Immunohistochemical staining of preserved surgical specimens was used. METHODS: Tissues from 12 patients with a histologic diagnosis of salivary MC (10 parotid, one submandibular gland, one minor salivary gland) were studied. Half (six of 12) of the patients had regional metastases. Tumor, normal salivary tissue, and regional lymph nodes were stained for HA, CD44, and HARE expression. Specimens were graded for staining intensity and a percent of the specimen stained. RESULTS: Normal salivary tissue did not demonstrate epithelial cell surface HA expression, whereas HA was expressed on tumor cells and in regional lymph nodes containing metastases. These differences were both significant using Student's t test (p < .00002, and p < .0022, respectively). Tumors with positive nodes tended to have greater cell surface HA. Decreased expression or downregulation of HARE was also noted in involved lymph nodes. No differences in CD44 expression were seen between primary specimens and lymph nodes. The observed staining patterns for CD44 and HARE were not reflective of the metastatic potential of the primary MC. CONCLUSIONS: Increased HA expression was seen on mucoepidermoid carcinoma cells compared with adjacent normal salivary gland epithelium. This observation may assist in explaining the development of regional metastasis in these tumors. We did not identify specific HA, CD44, or HARE staining patterns in primary lesions that were predictive of regional metastases. PMID- 16355389 TI - Effect of chemical composition on the induction of DNA damage by urban airborne particulate matter. AB - Airborne particulate matter (PM) contains a large number of genotoxic substances capable of endangering human health. In the present study, we have investigated the ability of chemically characterized water-soluble and organic-soluble fractions of two particle sizes (PM2.5 and PM10) from different regions of Mexico City to induce DNA damage in a human lung epithelial cell line. We also evaluated associations between the physicochemical parameters of the PM and its genotoxicity. The airborne particulate samples were collected from four regions of the city; a HiVol air sampler was used to collect PM10 on glass fiber filters and a tapered element oscillating system coupled to an automatic cartridge collection unit was used to collect PM2.5 on teflon filters. PM mass was determined by gravimetric analysis of the filters. Filters containing PM2.5 and one section of each PM10 filter were agitated either with deionized water to extract water-soluble compound, or with dichloromethane to prepare organic soluble compounds. The chemical composition of the extracts was determined by ion and gas chromatography and atomic adsorption spectroscopy. A549 human type II alveolar epithelial cells were exposed to different concentrations of the PM2.5 and PM10 extracts, and alkaline single cell gel electrophoresis or the Comet assay was performed to measure DNA damage and repair. These analyses indicated that soluble transition metals and the organic-soluble PM fractions are crucial factors in the DNA damage induced by PM. PM composition was more important than PM mass for producing genotoxicity. The results of this study showed that the constituents of the water-soluble PM extract are more likely to induce DNA damage than the organic compounds. PMID- 16355390 TI - Modulatory effects of Mentha piperita on lung tumor incidence, genotoxicity, and oxidative stress in benzo[a]pyrene-treated Swiss albino mice. AB - Mentha piperita or peppermint is currently used for alleviating nausea, flatulence, and vomiting. In the present investigation, we evaluated the chemopreventive, antigenotoxic, and antioxidative effects of an aqueous extract of Mentha piperita leaves. One-day-old Swiss albino mice were treated with a single subcutaneous injection of 0.5 mg benzo[a]pyrene (BP) and then given either water or a Mentha extract (ME; 1 g/kg body weight) by gavage starting at 3 weeks of age (weaning). The mice were killed at 9 weeks of age and tested for lung tumor incidence (chemoprevention); bone marrow micronucleus and chromosome aberration frequency (antigenotoxicity); and levels of liver and lung sulfhydral groups, superoxide dismutase (SOD) and catalase (CAT) activity, and lipid peroxidation (LPO) (antioxidative properties). The ME treatment resulted in a significant reduction in the number of lung adenomas from an incidence of 67.92% in animals given only BP to 26.31%, an inhibition of 61.26%. Tumor multiplicity was 1.22 in the BP-alone group and 1.15 in the BP + ME group. In addition, compared with the animals in the BP-alone group, ME reduced the frequency of chromosomal aberrations and micronuclei in bone marrow cells and decreased the levels of LPO and increased reduced glutathione content, and SOD and CAT activities in liver as well as lung. The results of this study indicate that ME is chemopreventive and antigenotoxic when given subsequent to an initiating dose of BP in newborn Swiss albino mice. The chemopreventive action and antigenotoxic effects observed in the present study may be due to the antioxidative properties of ME. PMID- 16355392 TI - Increased sensitivity to mitomycin C-induced sister chromatid exchange in lymphocytes from patients undergoing hyperbaric oxygen therapy. AB - Treatment of cells with hyperbaric oxygen (HBO) results in the generation of reactive oxygen species and the induction of DNA damage. In the present study, we have evaluated the sister chromatid exchange (SCE) frequencies in lymphocytes from patients undergoing hyperbaric oxygen therapy (HBOT). In addition, we have determined the sensitivity of lymphocytes from those patients to SCE induction by 20 and 40 ng/ml mitomycin C (MMC). Patients undergoing HBOT for diabetic feet were exposed to 10 consecutive daily HBO treatments according to a routine therapy protocol. The study began with 12 patients; however, it was not possible to sample all of the patients at all HBOT sessions, and the number of patients gradually decreased towards the end of the HBOT. We observed a statistically significant induction in mean SCE/cell (P < 0.05; n = 11) immediately after the first session of HBOT. Relative to its frequency after the 1st treatment, the mean SCE frequency gradually decreased after the 5th and 10th HBOT sessions and reached baseline (pretherapy) levels 1 day after the last treatment in the four patients that were sampled. The mean MMC-induced SCE frequency was highest in lymphocytes sampled immediately after the first HBOT session, and significantly higher than the MMC-induced SCE frequency in cells sampled before HBOT. Unlike the case with untreated cells, MMC-induced SCE frequencies remained high in lymphocytes sampled at later stages of therapy and mean MMC-induced SCE frequencies were significantly higher (P < 0.05; n = 4) in lymphocytes sampled 1 day after the last session of HBOT than in lymphocytes sampled from these patients prior to beginning the therapy. The results indicate that HBOT induces SCE and that lymphocytes retain increased sensitivity to the genotoxicity of MMC one day after completing the HBOT. PMID- 16355393 TI - Pseudogenes of the human HPRT1 gene. AB - Entrez Gene lists four HPRT1 gene pseudogenes (HPRTP1, HPRTP2, HPRTP3, and HPRTP4) mapping to chromosomes 3, 5, 11q, and 11q, respectively, as originally reported by Patel et al. in 1984 (Patel PI, et al. 1984 Somat Cell Mol Genet 10:483-493). However, the Entrez Gene reports for three of the four pseudogenes (HPRTP1, HPRTP3, and HPRTP4) are currently empty. A BLAST search of both GenBank (Homo sapiens) and the human genome found the chromosome 5 associated HPRTP2 sequence and a single chromosome 11q sequence (HPRTP3 or HPRTP4?). This chromosome 11 sequence had a unique 7.2 kb insert, which may explain why it originally appeared to be two separate pseudogenes. No evidence of a chromosome 3 associated sequence was found; however, a sequence highly homologous to HPRT1 was located on chromosome 4. All of these sequences are intronless processed pseudogenes. Lastly, a sequence highly homologous to HPRT1 exon 8 was found on chromosome 10. This homologous sequence was exactly exon 8 of a gene designated PRTFDC1, for phosphoribosyl transferase domain containing 1. This gene with unknown function is almost completely homologous to HPRT1 in exon structure (except for a 21 bp (seven amino acid) insertion in exon 1) and 68% homologous in amino acid sequence. PMID- 16355394 TI - Subacute thyroiditis: fine-needle aspiration cytology of 14 cases presenting with thyroid nodules. AB - Subacute thyroiditis (SAT) is usually diagnosed clinically without the need for fine-needle aspiration. The cytologic literature on this condition is therefore rare. We report on 14 cases of SAT presenting with thyroid nodules. The majority of patients were women with a mean age of 46 yr. All had pain/tenderness in the thyroid area accompanied by fever or an elevated ESR. The salient cytologic features included cellular smears; multinucleated giant cells in 100% of cases, some ingesting colloid or neutrophils; fibrous fragments with enmeshed inflammatory cells were a constant feature; follicular cells were scant to absent in most cases. Granulomas were rare. Colloid, when present was thick, with central cracks and frayed edges. One case was suspicious for malignancy. We conclude that the cytologic features of SAT are predictable, particularly, in the appropriate clinical setting. FNA is also helpful in ruling out concomitant neoplastic conditions. PMID- 16355395 TI - Cytologic comparison of a primary parathyroid cancer and its metastatic lesions: a case report. AB - We describe the fine-needle aspiration cytology features of a primary parathyroid cancer and of the local recurrent and distant metastatic lesions. The presence of prognostic factors Ki-67 and proliferating cell nuclear antigen (PCNA) was compared immunohistochemically between primary parathyroid carcinoma and related metastatic and recurrent foci. Flow cytometric DNA analysis was also performed to investigate any chromosomal abnormality of the parathyroid carcinoma. Cytologic examination of the endocrine tumor showed that it comprised a loose cohesive cluster and tumor cells with granular cytoplasm and mild nuclear atypia, but for purposes of cytodiagnosis, it is difficult to determine whether such a neoplasm is malignant on the basis of morphology alone. Immunohistochemical analysis showed that Ki-67 and PCNA labeling indices were higher in the recurrent and metastasized carcinomas than in the primary cancer, suggesting that neoplastic cells become more malignant in the recurrent and metastasized foci. To our knowledge, this is the first report describing not only cytopathologic but also immunocytologic differences between primary parathyroid cancer and the metastatic lesion. PMID- 16355396 TI - Cytologic differential diagnosis of follicular lymphoma grades 1 and 2 from reactive follicular hyperplasia: cytologic features of fine-needle aspiration smears with Pap stain and fluorescence in situ hybridization analysis to detect t(14;18)(q32;q21) chromosomal translocation. AB - Fine-needle aspiration cytology (FNAC) is a well-established technique for diagnosis of malignant lymphoma (ML). Generally, Giemsa but not Pap stain is used in FNAC. However, cytologic features obtained from Pap stain are also valuable. Very few studies on the cytologic characteristics of ML, as determined by Pap stain, are available. It is easier to observe nuclear irregularity and to identify nucleoli in ML cells by Pap stain than by Giemsa stain. Here, we applied Pap stain for cytomorphologic differential diagnosis of follicular lymphoma (FL) from reactive follicular hyperplasia (RFH). Eighteen biopsy-confirmed cases of FL grades 1 and 2, with available FNAC smears, and six cases of RFH were selected for this study. Low-power magnification showed well-known features, and tingible body macrophages and lymphoid cell aggregates were observed frequently in RFH and FL, respectively. In addition, the so-called two-nuclei-like cleaved cells were observed frequently in FL. These cells showed notably cleaved nuclei, and therefore, appeared to possess two nuclei. Under high-power magnification, the occurrence of cells with nucleoli >1 microm and of cleaved cells was high in FL compared to RFH. It is believed that FL derives from centrocytes and that FL cells are slightly larger than non-neoplastic small lymphocytes. However, analysis of cell diameter in this study indicated that small lymphoma cells were predominant in half the cases of FL grades 1 and 2, and the percentage of these cells was similar to that in RFH, showing why false-negative diagnosis of FL grades 1 and 2 occasionally occurs. There are limitations of FNAC in the diagnosis of FL. However, we believe that the appearance of two-nuclei-like cleaved cells and the high percentage of nucleoli-possessing cells, which we describe here, provide significant and valuable clues for the differential diagnosis of FL from RFH. Of 18 cases of FL grades 1 and 2, t(14;18)(q32;q21) was found in 13 cases with the use of destained FNAC smears. Our study suggests that, together with the cytomorphologic findings described earlier, FISH analysis for the chromosomal translocation, t(14;18)(q32;q21), is crucial for final cytologic diagnosis of FL grades 1 and 2. PMID- 16355397 TI - Fine-needle aspiration of a presacral myelolipoma. PMID- 16355398 TI - Interaction of erythrocytes with clue cells: light microscopic observation. PMID- 16355399 TI - Malignant biphasic pleural mesothelioma metastatic to the liver diagnosed by fine needle aspiration. AB - As malignant pleural mesotheliomas are most often rapidly fatal, distant metastases are rarely detected. Here, we report a unique case in which the diagnosis of metastatic pleural mesothelioma was made via cytologic examination of a fine-needle aspiration (FNA) of the liver. Recognition of the cytomorphologic features inherent to mesothelioma cells on FNA material may become important for proper patient management. To the best of our knowledge, the diagnosis of malignant pleural mesothelioma metastatic to the liver made by FNA has not been previously documented. PMID- 16355400 TI - Expression of peptidylarginine deiminase type 4 (PAD4) in various tumors. AB - Peptidylarginine deiminase type 4 (PAD4/PADI4) posttranslationally converts peptidylarginine to citrulline, in a process known as citrullination. Evidence suggests that PAD4 plays an essential role in pathogenesis of rheumatoid arthritis (RA). RA synovium has many features in common with tumor tissues, including abnormal cell proliferation, extensive fibrin deposition, high coagulation activity, and extreme angiogenesis. The purpose of the present study was to investigate expression of PAD4 in various tumor tissues. Immunohistochemistry indicated that PAD4 had significant expression in many tumor tissues, especially various adenocarcinoma. Western blotting with anti PAD4 antibody and immunostaining with anti citrulline antibody confirmed the expression of the enzyme in these tumors. Furthermore, our immunohistochemistry also detected co-location of PAD4 with cytokeratin (CK), a well-known tumor marker for oncological study in many tumors. Western blot analysis also detected citrulline signals in CK extracted from the tumors. In addition, CK 8, 18, and 19 following in vitro citrullination resisted to the digestion of caspase. The results further confirm the expression of PAD4 in the tumors and support that PAD4 may contribute to the disrupted apoptosis of tumors by caspase-mediated cleavage of CK. Double immunofluorescent labeling detected co-location of PAD4 with CD34, a cell marker of heamatopoietic progenitor cells (HPC) in bone marrow and other normal tissues, as well as in some fibroblast-like cells at stroma region of tumors, but not in the tumor cells. The findings imply that PAD4 is initially expressed in CD34(+) cells of bone marrow and then distributed in derives of the multi-potent progenitor cells in diverse tissues. The development of tumor cells expressing PAD4 is possibly associated with abnormal proliferation of CD34(+) stem cells. PMID- 16355401 TI - Differentiated rabbit prostatic stromal cells in primary culture display functional alpha1A-adrenoceptors. AB - AIMS: BPH is characterized by uncontrolled proliferation and increased contractility of prostatic smooth muscle cells. The activation of alpha1 adrenoceptors (alpha1-AR) seems involved in the latter event, but the lack of in vitro models expressing these receptors has hampered a more specific characterization of their role. In order to do so, we attempted to develop a new model of rabbit cultured prostatic stromal cells (PSC) in a non-proliferative and differentiated state. METHODS: The expression of cytoskeletal and stromal markers was confirmed by immunohistochemistry on primary cultured PSC. Alpha1-AR subtype expression was assessed by RT-PCR, while receptor coupling to the ERK1/ERK2 and calcium pathways was studied by Western Blot and Fura-2 calcium imaging, respectively. RESULTS: Cells grown under non-proliferative conditions displayed a differentiated phenotype, with expression of contractile cytoskeletal and stromal proteins. Furthermore, the alpha1A-AR was shown to activate ERK1/ERK2 as well as calcium signaling. CONCLUSION: These results emphasize the interest of this model for the characterization of PSC adrenergic regulation, in particular through the little-known alpha1A-AR. PMID- 16355402 TI - A two-step procedure for constructing confidence intervals of trait loci with application to a rheumatoid arthritis dataset. AB - Preliminary genome screens are usually succeeded by fine mapping analyses focusing on the regions that signal linkage. It is advantageous to reduce the size of the regions where follow-up studies are performed, since this will help better tackle, among other things, the multiplicity adjustment issue associated with them. We describe a two-step approach that uses a confidence set inference procedure as a tool for intermediate mapping (between preliminary genome screening and fine mapping) to further localize disease loci. Apart from the usual Hardy-Weiberg and linkage equilibrium assumptions, the only other assumption of the proposed approach is that each region of interest houses at most one of the disease-contributing loci. Through a simulation study with several two-locus disease models, we demonstrate that our method can isolate the position of trait loci with high accuracy. Application of this two-step procedure to the data from the Arthritis Research Campaign National Repository also led to highly encouraging results. The method not only successfully localized a well characterized trait contributing locus on chromosome 6, but also placed its position to narrower regions when compared to their LOD support interval counterparts based on the same data. PMID- 16355403 TI - Validity, efficiency, and robustness of a family-based test of association. AB - We propose a new test of linkage in the presence of allelic association that uses all available information in a sample of nuclear families, including parental phenotypes, genotypes from both affected and unaffected siblings, and families with homozygous parents. The test is based on the conditional framework developed by Rabinowitz and Laird [2000: Hum Hered 50:211-223] and is thus immune to population stratification and can be applied to families with any pattern of missing information. The test statistic is a conditional likelihood ratio based on a standard two-point linkage model with allelic association, where parameters are estimated from the sample. Through a simulation study, we determined that the proposed test has near optimal power for a wide range of scenarios, outperforming FBAT both when data were complete and when parental genotypes were missing, although differences between the two tests diminish as the genetic effect is reduced. To assess robustness, we also evaluated the performance of the tests under scenarios with population stratification and found that although there is a loss of efficiency, our proposed test remains a strong competitor to FBAT. PMID- 16355404 TI - Microsatellites versus Single-Nucleotide Polymorphisms in confidence interval estimation of disease loci. AB - With cost-effective high-throughput Single Nucleotide Polymorphism (SNP) arrays now becoming widely available, it is highly anticipated that SNPs will soon become the choice of markers in whole genome screens. This optimism raises a great deal of interest in assessing whether dense SNP maps offer at least as much information as their microsatellite (MS) counterparts. Factors considered to date include information content, strength of linkage signals, and effect of linkage disequilibrium. In the current report, we focus on investigating the relative merits of SNPs vs. MS markers for disease gene localization. For our comparisons, we consider three novel confidence interval estimation procedures based on confidence set inference (CSI) using affected sib-pair data. Two of these procedures are multipoint in nature, enabling them to capitalize on dense SNPs with limited heterozygosity. The other procedure makes use of markers one at a time (two-point), but is much more computationally efficient. In addition to marker type, we also assess the effects of a number of other factors, including map density and marker heterozygosity, on disease gene localization through an extensive simulation study. Our results clearly show that confidence intervals derived based on the CSI multipoint procedures can place the trait locus in much shorter chromosomal segments using densely saturated SNP maps as opposed to using sparse MS maps. Finally, it is interesting (although not surprising) to note that, should one wish to perform a quick preliminary genome screening, then the two-point CSI procedure would be a preferred, computationally cost-effective choice. PMID- 16355405 TI - The value of relatives with phenotypes but missing genotypes in association studies for quantitative traits. AB - The additional statistical power of association studies for quantitative traits was derived when ungenotyped relatives with phenotypes are included in the analysis. It was shown that the extra power is a simple function of the coefficient of additive genetic relationship and the phenotypic correlation coefficient between the genotyped and ungenotyped relatives. For close relatives, such as pairs of fullsibs and identical twin pairs, gains in power in the range of 10 to 30% are achieved if only one of the pair is genotyped. The theoretical results were verified by simulations. It was shown that ignoring the error in estimating the genotype of the ungenotyped relative has little impact on the estimates and on statistical power, consistent with results from quantitative trait loci (QTL) linkage studies. For genome-wide association studies in which not all relatives with phenotypes can be genotyped, our study provides a prediction of the additional power of an analysis that includes phenotypes on ungenotyped individuals, and can be used in experimental design. We show that a two-step procedure, in which missing genotypes are imputed and subsequently an association analysis is performed, is efficient and powerful. PMID- 16355406 TI - Comparison of methods incorporating quantitative covariates into affected sib pair linkage analysis. AB - For complex traits, it may be possible to increase the power to detect linkage if one takes advantage of covariate information. Several statistics have been proposed that incorporate quantitative covariate information into affected sib pair (ASP) linkage analysis. However, it is not clear how these statistics perform under different gene-environment (G x E) interactions. We compare representative statistics to each other on simulated data under three biologically-plausible G x E models. We also compared their performance with a model-free method and with quantitative trait locus (QTL) linkage approaches. The statistics considered here are: (1) mixture model; (2) general conditional logistic model (LODPAL); (3) multinomial logistic regression models (MLRM); (4) extension of the maximum-likelihood-binomial approach (MLB); (5) ordered-subset analysis (OSA); and (6) logistic regression modeling (COVLINK). In all three G x E models, most of these six statistics perform better when using the covariate C1 associated with a G x E interaction effect than when using the environmental risk factor C2 or the random noise covariate C3. Compared with a model-free method without covariates (S(all)), the mixture model performs the best when using C1, with the high-to-low OSA method also performing quite well. Generally, MLB is the least sensitive to covariate choice. However, most of these statistics do not provide better power than S(all). Thus, while inclusion of the "correct" covariate can lead to increased power, careful selection of appropriate covariates is vital for success. PMID- 16355407 TI - Directed evolution of the epidermal growth factor receptor extracellular domain for expression in yeast. AB - The extracellular domain of epidermal growth factor receptor (EGFR-ECD) has been engineered through directed evolution and yeast surface display using conformationally-specific monoclonal antibodies (mAbs) as screening probes for proper folding and functional expression in Saccharomyces cerevisiae. An EGFR mutant with four amino acid changes exhibited binding to the conformationally specific mAbs and human epidermal growth factor, and showed increased soluble secretion efficiency compared with wild-type EGFR. Full-length EGFR containing the mutant EGFR-ECD was functional, as assayed by EGF-dependent autophosphorylation and intracellular MAPK signaling in mammalian cells, and was expressed and localized at the plasma membrane in yeast. This approach should enable engineering of other complex mammalian receptor glycoproteins in yeast for genetic, structural, and biophysical studies. PMID- 16355410 TI - Intact surface of bioactive glass S53P4 is resistant to osteoclastic activity. AB - Bioactive glass reacts with body fluids and is gradually dissolved in tissues and in cell cultures. We investigated whether osteoclasts contribute to this process, by culturing newborn rat bone-marrow cells containing osteoclasts on polished bioactive glass plates (glass S53P4). The cultures were inspected at days 1-5 and stained for alkaline phosphatase (ALP) to demonstrate osteoblasts and for tartrate resistant acid phosphatase (TRAP) to visualize osteoclasts. Nonosteoclastic cells proliferated several-fold both on bioactive glass and on plastic, whereas osteoclasts and their precursors matured into multicellular giant cells and degenerated. Most cells on bioactive glass became ALP-positive, whereas on plastic the majority of cells remained ALP-negative. Osteoclasts survived on bioactive glass for 4-5 days, whereas on plastic they degenerated and disappeared after 3 days. Condensed nuclei indicating apoptosis were detected both in degenerating osteoclasts and osteoblasts. The surface of the bioactive glass reacted rapidly forming rounded pits, erosions, and cracks within 24 h in areas occupied by osteoblasts. Light microscopy and scanning electron micrographs demonstrated, however, a smooth surface below the cytoplasm of osteoclasts. This indicates that when applied on an intact bioactive glass surface, osteoclasts were unable to dissolve the glass material within this culture period. PMID- 16355408 TI - Experimental investigation of interfaces in hydroxyapatite/polyacrylic acid/polycaprolactone composites using photoacoustic FTIR spectroscopy. AB - Molecular interactions in hydroxyapatite (HAP) polymer composites have been studied using photoacoustic spectroscopy. HAP is mineralized by wet precipitation under two conditions: first is in the absence of polyacrylic acid (PAAc) (ex-situ HAP) and second in the presence of PAAc (in-situ HAP). Porous and solid composites of ex-situ and in-situ HAP with polycaprolactone (PCL) have also been made to evaluate their applicability as bone scaffolds. Photoacoustic Fourier transform infrared (PA-FTIR) spectroscopy studies indicate that both in-situ and ex-situ HAP have HPO4 (2-) in their structure, which leads to Ca2+ deficiency. During crystallization of in-situ HAP, PAAc dissociates to form carboxylate ions, which binds to calcium ions and act as suitable site for nucleation for HAP crystallization. PA-FTIR spectra of porous and solid composites indicate that porous composites adsorb more water, which is hydrogen bonded with carbonyl of PCL. Mechanical tests on solid samples indicate that ex-situ HAP/PCL composites have higher elastic modulus than in-situ HAP/PCL composites. However, in case of porous composites, in-situ HAP/PCL composites are found to have higher elastic modulus. In-situ HAP is chemically and structurally different from ex-situ HAP. This modified HAP causes variation in microstructure of porous composite and hence alteration of its load transfer mechanisms and hence mechanical properties. PMID- 16355411 TI - Bone morphogenetic proteins 5 and 6 stimulate osteoclast generation. AB - Bone regeneration is required for fracture-healing, and different procedures have been used to promote osteogenesis. Recently, BMP-2 has been shown to induce bone formation in vivo and has been tested in clinical trials. A recent in vitro study evaluated the osteogenic activity of 14 BMPs on osteoblastic progenitor cells with an osteogenic hierarchical model in which BMP-2 and BMP-6 may play an important role in inducing osteoblast differentiation. Although the relative osteoinductive activity of each BMP is important, bone regeneration is a process consisting of bone formation and bone resorption. Therefore, it remains unclear which effects BMP-5 and -6 have on the generation of osteoclasts and by which mechanism osteoclastogenesis is stimulated. To compare osteoclastic potency of each BMP, primary murine bone marrow cells were treated with human recombinant BMP-2, BMP-5, or BMP-6 and 1,25-(OH)2 vitamin D3 and stained for the TRAP enzyme. Osteogenic activity of BMP-5 was determined by measuring induction of ALP activity and proliferation after incubation with primary murine osteoblasts. For elucidating the molecular mechanism, primary bone marrow cells with various concentrations of OPG were added to the TRAP assay and mRNA levels of RANKL and OPG were measured after stimulation with BMP-5. The presented data show that BMP 5 and BMP-6, unlike BMP-2, enhanced the formation of murine TRAP+/MNCs in a biphasic curve. BMP-5 and -6 were less potent in stimulating osteoclastogenesis compared to BMP-2. Concerning the effects of BMP-5 on osteoblasts, there was a dose-dependent increase of ALP activity and proliferation up to a maximum dose of 300 ng/mL. At the mRNA level, BMP-5 increased the RANKL/OPG ratio. In conclusion, this study demonstrates that in contrast to BMP-2, BMP-5 and -6 influences the generation of osteoclasts in a biphasic mode. Both proteins might be very important regulators of bone homeostasis, and therefore, potent candidates for future treatment strategies of bone regeneration. PMID- 16355412 TI - Effects of a biodegradable polymer synthesized with inorganic tin on the chondrogenesis of human articular chondrocytes. AB - Recent study has shown that biodegradable polymers are attractive candidates for chondrocyte fixation and further transplantation in cartilage tissue engineering. Poly (glycolic acid) (PGA), a polymer of glycolic acid, is widely used in orthopedic applications as a biodegradable polymer. Organotin, lead, antimony, and zinc are catalysts commonly used in synthesizing PGA. Here, we investigated the biocompatibility of PGA, synthesized with and without inorganic tin as a catalyst in chondrogenesis of human articular chondrocytes in a micromass culture system. Significant enhancement of chondrocyte proliferation and expression of the collagen type II protein gene were observed in cultures treated with PGA synthesized with a tin catalyst. However, aggrecan gene expression was very similar to the control culture. Amount of collagen type II protein was also increased in the same group of cultured chondrocytes. In contrast, PGA without a catalyst caused overall inhibition of chondrogenesis. Despite several positive findings, extensive investigations are essential for the feasibility of this PGA(Sn) in future clinical practice. PMID- 16355413 TI - Three-dimensional culture of porcine fetal liver cells for a bioartificial liver. AB - A three-dimensional (3-D) culture experiment of porcine fetal liver cells (FLCs) was performed using a porous resin substrate, for the purpose of developing a bioartificial liver. A long-term 3-D culture and monolayer culture as the control were performed for more than 1 month. To promote cell growth and maturation, human oncostatin M (OSM), the human leukemia inhibitory factor (LIF), or cortisol was added to the cultures, and the effect of each agent on cell proliferation and liver-specific cellular functions was investigated. The cell numbers in both the monolayer and 3-D cultures increased gradually with time, irrespective of the supplementation of the stimulating agents. In the monolayer culture, the albumin secretion of FLCs decreased rapidly, and scarce activity was detected from 2 weeks onward under all culture conditions tested. In the 3-D cultures, neither human OSM nor human LIF had any definite effect on the albumin secretion of FLCs. However, in the cultures with cortisol, albumin secretion was maintained for a considerably long period. These findings suggest that a bioartificial liver can be developed by culturing porcine FLCs with cortisol as the stimulant. PMID- 16355414 TI - Solvent participation in Serratia marcescens endonuclease complexes. AB - The monomer and dimer of the bacterium Serratia marcescens endonuclease (SMnase) are each catalytically active and the two subunits of the dimer function independently of each other. Specific interfacial waters may play a role in stability, complex formation, and functionality. We performed molecular dynamics simulations of both a subunit of SMnase and its model built complex with DNA and analyzed the relation of the hydration sites to the catalytic mechanism. It was found that the binding of DNA has little influence on the global hydration properties of the protein, including occupancy and water residence time distributions. DNA and protein recognition in our model mainly involves direct contacts by hydrogen bond and hydrophobic interactions. Water-mediated contacts exist, but are less common. Three interior water clusters were identified for SMnase. One cluster around the active site in the monomer-DNA complex shows relatively strong interactions between hydration sites as well as between the sites and the biomolecules. The simulated cluster properties agreed well with experimental data. The magnesium ion shows ligand exchange. Although Mg2+ keeps six ligands during the entire simulation, upon the binding of DNA, Asn119 loses its coordination with Mg2+, while one nonbridging oxygen of the phosphate of a DNA residue and two oxygen atoms of the side chain of Glu127 become the ligands. Waters in a nearby cluster exchange and participate in the resolvation of groups in the presence of DNA. Water thus not only participates in the cleavage of DNA but also can stabilize the transition state and the leaving groups in our model. PMID- 16355415 TI - Conversion of type I 4:6 to 3:5 beta-turn types in human acidic fibroblast growth factor: effects upon structure, stability, folding, and mitogenic function. AB - Human acidic fibroblast growth factor (FGF-1) is a member of the beta-trefoil superfold, a protein architecture that exhibits a characteristic threefold axis of structural symmetry. FGF-1 contains 11 beta-turns, the majority being type I 3:5; however, a type I 4:6 turn is also found at three symmetry-related locations. The relative uniqueness of the type I 4:6 turn in the FGF-1 structure suggests it may play a key role in the stability, folding, or function of the protein. To test this hypothesis a series of deletion mutations were constructed, the aim of which was to convert existing type I 4:6 turns at two locations into type I 3:5 turns. The results show it is possible to successfully substitute the type I 4:6 turn by a type I 3:5 turn with minimal impact upon protein stability or folding. Thus, these different turn structures, even though they differ in length, exhibit similar energetic properties. Additional sequence swapping mutations within the introduced type I 3:5 turns suggests that the turn sequence primarily affects stability but not turn structure (which appears dictated primarily by the local environment). Although the results suggest that a stable, foldable beta-trefoil protein may be designed utilizing a single turn type (type I 3:5), a type I 4:6 turn at turn 1 of FGF-1 appears essential for efficient mitogenic function. PMID- 16355416 TI - Generalized correlation for biomolecular dynamics. AB - Correlated motions in biomolecules are often essential for their function, e.g., allosteric signal transduction or mechanical/thermodynamic energy transport. Because correlated motions in biomolecules remain difficult to access experimentally, molecular dynamics (MD) simulations are particular useful for their analysis. The established method to quantify correlations from MD simulations via calculation of the covariance matrix, however, is restricted to linear correlations and therefore misses part of the correlations in the atomic fluctuations. Herein, we propose a general statistical mechanics approach to detect and quantify any correlated motion from MD trajectories. This generalized correlation measure is contrasted with correlations obtained from covariance matrices for the B1 domain of protein G and T4 lysozyme. The new method successfully quantifies correlations and provides a valuable global overview over the functionally relevant collective motions of lysozyme. In particular, correlated motions of helix 1 together with the two main lobes of lysozyme are detected, which are not seen by the conventional covariance matrix. Overall, the established method misses more than 50% of the correlation. This failure is attributed to both, an interfering and unnecessary dependence on mutual orientations of the atomic fluctuations and, to a lesser extent, attributed to nonlinear correlations. Our generalized correlation measure overcomes these problems and, moreover, allows for an improved understanding of the conformational dynamics by separating linear and nonlinear contributions of the correlation. PMID- 16355417 TI - Reversible constriction of the fetal ductus arteriosus after maternal use of topical diclofenac and methyl salicylate. PMID- 16355418 TI - Diabetes preventive gluten-free diet decreases the number of caecal bacteria in non-obese diabetic mice. AB - BACKGROUND: A gluten-free diet reduces the incidence of diabetes mellitus in non obese diabetic (NOD) mice, but the mechanism is not known. The aim of this study was to examine the possible influence of the diet on the caecal bacterial flora, which may affect the intestinal physiology and mediate disease prevention. METHODS: Two groups of NOD mice from the age of 3 weeks were fed either a gluten free diet or a standard diet. Each diabetic mouse, when diagnosed, along with a non-diabetic mouse from the same diet group and two non-diabetic mice from the alternate diet group were euthanized and sampled for classical bacteriological examination. RESULTS: Nine out of 19 (47%) standard-fed mice and 1 out of 19 (5%) gluten-free-fed mice developed diabetes (p < 0.01). Mice on the gluten-free diet had significantly fewer aerobically (p < 0.01) and microaerophilically (p < 0.001) cultivated bacteria in their intestines than standard-fed mice. Non diabetic mice also had significantly fewer microa erophilic and anaerobic bacteria than diabetic mice (p < 0.05). These differences were primarily due to a difference in the Gram-positive flora. CONCLUSIONS: The gluten-free diet compared to the standard diet both qualitatively and quantitatively substantially altered the composition of the caecal bacterial flora in NOD mice. Although Gram-positive bacteria might influence the beta cells through certain digestive products, it is more likely to assume that any effect on diabetes incidence is immunological. PMID- 16355419 TI - The relevance of international consortia in studies on the pathogenesis of type 1 diabetes: the role of ET1DGN. AB - The Type 1 Diabetes Genetics Consortium (T1DGC), funded September 2002, is a collaborative effort to develop resources for the purpose of identifying risk genes for type 1 diabetes. One of the aims is to establish a renewable resource of DNA on 2,500 affected sib-pair families. Regional networks have been established and in continental Europe, the European Type 1 Diabetes Genetics Network (ET1DGN) is responsible for identifying and collecting families. Most European countries have shown interest in this study and many have started recruiting families. The consortium is open to participation and there are several ways one can participate as a member of the consortium. ET1DGN suggests going beyond the tasks of T1DGC and establishing a well-organized network for additional and future studies in Europe. PMID- 16355420 TI - Epitope mapping of the protective antigen of B. anthracis by using nanoclusters presenting conformational peptide epitopes. PMID- 16355421 TI - Separation of hydrocarbons with a microporous metal-organic framework. PMID- 16355422 TI - Functionalized benzylic magnesium reagents through a sulfur-magnesium exchange. PMID- 16355423 TI - The importance of inter- and intramolecular van der Waals interactions in organic reactions: the dimerization of anthracene revisited. PMID- 16355424 TI - Helicity induction in hydrogen-bonding-driven zinc porphyrin foldamers by chiral C60-incorporating histidines. PMID- 16355425 TI - Virus-engineered colloidal particles--a surface display system. PMID- 16355426 TI - Ligand-selective aqueous synthesis of one-dimensional CdTe nanostructures. PMID- 16355427 TI - A highly active, heterogeneous catalyst for alkyne metathesis. PMID- 16355428 TI - Ordered nanostructures from the self-assembly of reactive coil-rod-coil molecules. PMID- 16355429 TI - Total synthesis of (-)- and ent-(+)-vindorosine: tandem intramolecular Diels Alder/1,3-dipolar cycloaddition of 1,3,4-oxadiazoles. PMID- 16355430 TI - Computational design of a human butyrylcholinesterase mutant for accelerating cocaine hydrolysis based on the transition-state simulation. PMID- 16355431 TI - The [Ga2(C5Me5)]+ ion: bipyramidal double-cone structure and weakly coordinated, monovalent Ga+. PMID- 16355432 TI - Particle-stabilized emulsions: a bilayer or a bridging monolayer? PMID- 16355433 TI - Reagent-controlled switching of 5-exo to 6-endo cyclizations in epoxide openings. PMID- 16355434 TI - Tetraanionic organoborate squares glued together by cations to generate nanotubular stacks. PMID- 16355435 TI - Depression in paediatric cancer: an overview. AB - Research into depression in paediatric cancer is in its early stages, but nevertheless has presented interesting challenges regarding the recognition and measurement of depression in a medically ill population. In this article we discuss the complex interaction between physical and psychological variables, and the diagnostic difficulties arising from this. We review the epidemiological findings regarding prevalence, evaluating the apparently low prevalence rate in the light of methodological weaknesses. Hypotheses put forward to explain the findings are discussed. We conclude by highlighting areas for future research. PMID- 16355436 TI - The versatility of pentalene coordination to transition metals: a density functional theory investigation. AB - DFT calculations with full geometry optimization have been carried out on a series of real and hypothetical compounds of the type [CpM(C8H6)], [(CO)3M(C8H6)], [M(C8H6)2], [(CpM)2(C8H6)], [[(CO)3M]2(C8H6)], and [M2(C8H6)2] (M = transition metal). The bonding in all the currently known compounds is rationalized, as well as in the (so far) hypothetical stable complexes. Depending on the electron count and the nature of the metal(s), eta2 (predicted), eta3, eta5, eta8, or intermediate coordination modes can be adopted. In the case of the mononuclear species, the most favored closed-shell electron counts are 18 and 16 metal valence electrons (MVE). In the case of the dinuclear species, an electron count of 34 MVEs is most favored. However, other electron counts can be stabilized, especially in the case of dinuclear complexes. Coordinated pentalene should most often be considered as formally being a dianion, but sometimes as a neutral ligand. In the former case it can behave as an aromatic species made of two equivalent fused rings, as a C5 aromatic ring connected to an allylic anion, or even as two allylic anions bridged by a C7=C8 double bond. In the latter case, it can behave as a bond-alternating cyclic polyene or as a C5 aromatic ring connected to an allylic cation. PMID- 16355437 TI - Amplification of chirality in hydrogen-bonded tetrarosette helices. AB - The amplification of chirality in hydrogen-bonded tetrarosette assemblies under thermodynamic equilibrium is described. The extent of the chiral amplification obtained by means of "sergeants-and-soldiers" experiments depends only on the structure of the assembly and it is independent of the methodology used for the formation of the tetrarosette assemblies. The difference in free energy (deltaG(o)(M/P)) between the M- and P-diastereomeric helices is up to 40 times higher than for double rosette assemblies. PMID- 16355438 TI - Interaction of indole derivatives and tryptophan peptides with interfaces of sodium dodecyl sulfate micelles. AB - The free energies of transfer for indole and tryptophan derivatives and pentapeptides having single tryptophan residues from aqueous to sodium dodecyl sulfate (SDS) micellar phases have been systematically studied using the conventional method of ultraviolet absorption spectrophotometry. The free energies for the position isomers of methyl indoles varied depending on the substitution positions. Thus, the contribution of the methyl group to the binding affinity of the 4-methyl indole to the micelle was about twice that of the 2- and 7-methyl indoles. The free energy changes with the introduction of halogen groups to the indole rings were correlated to the nonpolar water-accessible surface area (DeltaA(np)) of the halogen moieties, which were regarded as hydrophobic. The relationships followed straight lines passing through the origins. Position dependence having tendencies similar to the methyl indoles was observed among the magnitudes of the slopes of the straight lines. These results strongly suggest that the indole rings of the derivatives residing in the micellar interface regions direct their imino moieties --NH-- toward the micellar surfaces. Experiments using model tryptophan pentapeptides showed that the magnitude of free energy change per methylene unit of an alkyl amino acid residue in the pentapeptide increased with elongation of the alkyl moiety and was not a constant value as reported for various alkyl compounds. When the peptides distribute to the SDS micelles, the peptide backbones are anchored in aqueous phases and the amino acid side chains in the interfaces extend their alkyl groups toward the micellar centers. Thus, the free energy changes can be connected to the positions of the alkyl groups of the amino acid residues in the micelles. PMID- 16355439 TI - Using internal and collective variables in Monte Carlo simulations of nucleic acid structures: chain breakage/closure algorithm and associated Jacobians. AB - This article describes a method for solving the geometric closure problem for simplified models of nucleic acid structures by using the constant bond lengths approximation. The resulting chain breakage/closure equations, formulated in the space of variable torsion and bond angles, are easy to solve, and have only two solutions. The analytical simplicity is in contrast with the high complexity of the closure problem in the torsion angle space with at most 16 solutions, which has been dealt with by several authors and was solved analytically by Wu and Deem (J. Chem. Phys. 1999, 111, 6625). The discussion on the choice of variables and associated Jacobians is focussed on the question of how conformational equilibration is affected in Monte Carlo simulations of molecular systems. In addition to the closure of the phosphate backbone, it is necessary to also solve the closure problem for the five-membered flexible furanose sugar ring. Explicit closure equations and the resulting Jacobians are given both for the complete four-variable model of the furanose ring and simulations in the phase-amplitude space of the five-membered ring, which are based on the approximate two-variable model of furanose introduced by Gabb et al. (J. Comput. Chem. 1995, 16, 667). The suggested closure algorithm can be combined with collective variables defined by translations and rotations of the monomeric nucleotide units. In comparison with simple internal coordinate moves, the resulting concerted moves describe local structural changes that have high acceptance rates and enable fast conformational equilibration. Appropriate molecular models are put forward for prospective Monte Carlo simulations of nucleic acids, but can be easily adapted to other biomolecular systems, such as proteins and lipid structures in biological membranes. PMID- 16355440 TI - Pharmacokinetic of rhein in healthy male volunteers following oral and retention enema administration of rhubarb extract: a single dose study. AB - Rhubarb is a common herb used in traditional Chinese medicine. However, few publications exist about its pharmacokinetic profiles in animals or healthy volunteers. Whether retention enema administration of rhubarb extract affects its pharmacokinetics as well as its tolerability is unknown. Therefore, we set out to compare the pharmacokinetic parameters of rhein administered by retention enemas with those of conventional oral dosing of rhubarb extract. Eight healthy male volunteers were enrolled in a prospective crossover study. All subjects received a single dose of rhubarb extract (50 mg.kg(-1)) on two separate occasions, once orally, once by a retention enema. Rhein plasma concentration was measured by HPLC. The C(max), AUC(0-infinity), AUMC were significantly higher in oral administration than those in retention enema administration (p < 0.05), while V(d) of rhein after oral administration of rhubarb extract was significantly lower (p < 0.05) than that after retention enema administration. However, no statistically significant differences between the two treatments were observed for any of the other pharmacokinetic parameters (T(max), t(1/2), MRT(0-infinity), CL). Dosage adjustment is advisable for retention enema administration of rhubarb extract in patients. PMID- 16355441 TI - Comparison of body composition methods during weight loss in obese women using herbal formula. AB - Bioelectrical impedance analysis (BIA), a device that analyzes the current conduction differences between the fat and water components is widely used for reasons that include convenience of use, non-invasiveness, safety, and low cost. Dual energy X-ray absorptiometry (DXA) allows for the assessment of total body and regional lean and fat tissues and bone mineral content (BMC). The objective of this study was to compare body composition assessments by BIA and DXA before and after a 6-week herbal diet intervention program in 50 pre-menopausal women [mean +/- SD: age 30.58 +/- 6.15, body mass index (BMI) 31.72 +/- 3.78]. Waist-to hip ratio (WHR) was measured by BIA and anthropometry. Lean body mass (LBM), body fat (BF), BMC and percent body fat (%BF) were measured by BIA and DXA. Highly significant correlations were observed between BIA and DXA measurements for LBM, BF, BMC and %BF (r = 0.73, 0.93, 0.53, 0.79, respectively) before the intervention. Differences between BIA and DXA measurements were observed in LBM, BF, %BF and BMC before intervention (p < 0.01) where WHR by BIA was significantly higher compared to anthropometry before (p < 0.01) and after the intervention (p < 0.01). BIA underestimated LBM by 1.85 kg and overestimated BF by 2.54 kg compared to DXA before the intervention. Although BIA and DXA showed highly significant correlations for LBM, BF, BMC and %BF before the intervention, they did not produce statistically comparable results in pre-menopausal Korean women and therefore should not be used interchangeably when measuring body composition. PMID- 16355442 TI - Computerized diagnosis from tongue appearance using quantitative feature classification. AB - This study investigates relationships between diseases and the appearance of the human tongue in terms of quantitative features. The experimental samples are digital tongue images captured from three groups of candidates: one group in normal health, one suffering with appendicitis, and a third suffering with pancreatitis. For the purposes of diagnostic classification, we first extract chromatic and textural measurements from original tongue images. A feature selection procedure then identifies the measures most relevant to the classifications, based on which of the three tongue image categories are clearly separated. This study validates the use of tongue inspection by means of quantitative feature classification in medical diagnosis. PMID- 16355443 TI - Relationship between bone mineral density and syndrome types described in traditional chinese medicine in chronic obstructive pulmonary disease: a preliminary clinical observation. AB - Osteoporosis is a common finding following chronic obstructive pulmonary disease (COPD), but there are few reports on the relationship between bone mineral density (BMD) and the syndrome types described in traditional Chinese medicine (TCM) in patients with COPD. A cross-sectional medical survey was used in this study. Twenty-six male patients with COPD and 26 age-matched male healthy subjects were recruited. The symptom questionnaire survey of TCM was implemented, and thereafter the COPD patients were divided into two subgroups: type of deficiency of the lung and spleen (TDLS) and type of deficiency of the lung, spleen and kidney (TDLSK). BMD of lumbar spine (L2-4), non-dominant femoral neck (Neck), Ward's triangle (Ward's), and great trochanter (Troch) were measured by dual-energy x-ray absorptiometry. In addition, the other bone turnover markers were also examined. The results showed that BMD was much more decreased in TDLSK than that in TDLS patients (p < 0.05), and BMD in the patients of the TDLS subgroup without symptoms of kidney-vacuity has showed the decreased trend from healthy subjects to TDLS patients. Furthermore, there was a higher incidence of osteoporosis in patients with TDLSK compared with that in TDLS (p < 0.05, OR > 2.0). Therefore, the data suggest that: (1) BMD might be a marker more sensitive than the symptom for the diagnosis of kidney-vacuity in COPD patients; (2) the deficiency of kidney would be the key factor of bone mineral loss; and (3) that invigorating the kidney should be performed in the phase of TDLS in COPD patients in advance. PMID- 16355444 TI - Effects of acupuncture on potential along meridians of healthy subjects and patients with gastric disease. AB - The purpose of this study was to investigate the differences in bio-potential in the stomach meridians of 20 normal healthy control subjects (NHC) and 20 patients with gastric disease (PGD). We measured changes in potential between ST-39 and ST 37 on the stomach meridian in response to invasive insertion of a needle at ST 36. The response patterns of NHC were regular in both the left and right meridians, whereas the response patterns of PGD were irregular. The potential of the NHC was significantly higher than that of the PGD group. The potential values in the left and right meridians of NHC were the same. However, the potential in the right meridian was significantly higher than that in the left meridian of PGD (p < 0.001). This study indicates that there may be changes in Qi or in the electrical properties of meridians according to health status, reflected in the regularity of the response to acupuncture or in the level of potential along meridians. Therefore, it may be possible to make a diagnosis from the state of a meridian by comparing levels of potential difference and its regularity. PMID- 16355445 TI - Acupuncture reduces alcohol withdrawal syndrome and c-Fos expression in rat brain. AB - Acupuncture as a therapeutic intervention is widely practiced in the treatment of many functional disorders including alcohol abuse. In the present study, the effects of acupuncture on alcohol withdrawal syndrome (AWS) and Fos-like immunoreactivity (FLI) in the striatum and the nucleus accumbens (NAC) of rats were investigated. During 3 days of cessation following chronic administration of ethanol (3 g/kg, i.p. for 3 weeks), rats showed a significant increase in AWS, such as hypermotility, tail rigidity, grooming and tremor, and an increase in FLI in the dopamine terminal areas of the brain. Treatment with acupuncture at zusanli (ST36) or sanyinjiao (SP6) during the withdrawal period inhibited both AWS and FLI of rats undergoing ethanol injection. These results suggest that acupuncture may be useful in the treatment of alcoholism by modulating post synaptic neural activation in the striatum and NAC. PMID- 16355446 TI - Building a bridge for integrating Chinese medicine into conventional healthcare: observations drawn from the development of the Chinese quality of life instrument. AB - This paper describes the methods and theories of patient-reported outcomes, in particular to the health-related quality of life recently applied in Chinese medicine research. It begins with an investigation of the reasons for a patient reported outcomes measure for Chinese medicine and the development of a new health-related quality of life instrument based on Chinese culture and Chinese medicine. Discussions on the importance and application of patient-reported outcomes as well as the relationship between quality of life and Chinese medicine are at the focus of this paper. Through a description of the Chinese Quality of Life Instrument and its development, the present work demonstrates an evidence based approach using patient-reported outcomes or health-related quality of life measures to evaluate treatment efficacy of Chinese medicine, and thereby build a bridge for the integration of Chinese medicine into mainstream health care. PMID- 16355447 TI - A birdcage model for the Chinese meridian system: Part V: Applications to animals and plants. AB - Since we all belong to the Kingdom Animalia, it is not surprising that animals in general benefit from the healing art of acupuncture that helps humans. Consequently, any proposed mechanism of Qi and acupuncture for humans based on animal physiology is probably applicable to animals as well, yet none is capable of explaining most of the complicated physiological effects observed. Not much attention was paid to the effects of Qi and acupuncture on plants (Kingdom Plantae) and on enoki mushrooms (Kingdom Fungi) by the TCM community, probably because they cannot be explained in terms of neurochemistry or connective tissue structures. However, our transmission and birdcage model is in principle applicable across Kingdom boundaries, because it is based on physical properties underlying the biological structure, thus its explanatory power is not restricted by categories of biology. We estimate several possible parameters of the birdcage model for animals and plants and give a possible interpretation for the sound fertilization phenomenon. PMID- 16355448 TI - Anti-hypertension effects of traditional Chinese medicine ju-ling-tang on renal hypertensive rats. AB - This research investigated the anti-hypertension effect of the traditional Chinese medicine (TCM) ju-ling-tang (JLT) on an animal model of hypertension induced by unilateral renal artery ligation. In the study of anti-hypertension effects, 60 minutes after oral administration with NG tube feeding of 240 mg/kg JLT, a significant decrease in blood pressure (p < 0.05) was observed and sustained till 120 minutes. In the group given 50 mg/kg alpha-methyldopa orally, the effect was obvious 90 minutes after medication (p < 0.01), and lasted until 240 minutes. In terms of organ pathology, a significant reduction in the extent of induced glomerular sclerosis was observed in rats given 240 mg/kg JLT compared with the control. From these results, we infer that JLT has a beneficial anti hypertensive effect on renal hypertension. PMID- 16355450 TI - Pharmacokinetics of berberine in rat thalamus after intravenous administration of Coptidis rhizoma extract. AB - A rapid and simple high-performance liquid chromatographic method for determination of berberine in rat thalamus was described in this study. Thalamus samples were pretreated by protein precipitation with methanol and acetonitrile. Berberine was determined using a Hypersil C(18) column with an isocratic mobile phase of acetonitrile--0.05 M potassium dihydrogen phosphate (containing 0.5% triethylamine, pH 3.0) (30:70 v/v) and with UV detection at 265 nm. The lower limit of quantification for berberine in thalamus was 24 ng/ml, and the lowest concentration of berberine determined in rat thalamus samples was 47.5 ng/ml at 48 hours. The calibration curve for berberine was linear (r(2) = 0.9994) over the concentration range 24-6000 ng/ml. At this concentration range, the overall recoveries (91.20%-93.24%) for berberine were determined and the accuracy of the intra- and inter-day assays from rat thalamus were less than 6% RSD. Following intravenous administration of 10.2 mg/kg of Coptidis Rhizoma (CR) extract containing 3 mg/kg berberine into rats, the thalamus level of berberine increased rapidly (t(1/2alpha) = 1.93 hours), peaked at 2.48 hours with a concentration of 271 ng/g, and had a slow elimination rate (t(1/2beta) = 14.6 hours), which suggested that berberine might directly act on certain regions of the thalamus, have pharmacological effects on some cerebral dysfunctions, and be an active ingredient of huang lian jie du tang for the treatment of cerebral disease. PMID- 16355449 TI - Effects of Gingko biloba extract on gap junction changes induced by reperfusion/reoxygenation after ischemia/hypoxia in rat brain. AB - Gap junction communication between astrocytes plays an important role in the brain. The purpose of this study was to investigate the effects of Gingko biloba extract (GBE) on the changes of connexin 43 (Cx43) mRNA and protein expression levels of rat cortex and hippocampus induced by ischemia-reperfusion and astrocyte gap junction intercellular communication (GJIC) induced by hypoxia reoxygenation. After 2 hours of middle cerebral artery occlusion (MCAO) followed by 24 hours of reperfusion, there was obvious neurological deficit in rats. Cx43 mRNA and protein expression levels of rat cortex and hippocampus in the ischemia hemisphere were decreased significantly. When GBE at doses of 50 and 100 mg/kg body weight was administrated by p.o. daily for 7 days, the neurological deficit was improved, and lower Cx43 mRNA and protein expression levels induced by ischemia-reperfusion were recovered to normal. The i.p. injection of nimodipine (0.7 mg/kg weight body) also showed improvement on neurological deficit and Cx43 expression levels. Astrocyte GJIC was measured by the fluorescence recovery after photobleaching (FRAP). Hypoxia-reoxygenation induced a significant decrease in GJIC. Pretreatment with GBE (100 mg/l) and nimodipine (1.6 mg/l) significantly prevented the hypoxia-reoxygenation inhibition of GJIC. These results suggest that GBE could exert its neuroprotective effects by improvement of Cx43 expression and GJIC induced by hypoxia/ischemia-reoxygenation/ reperfusion injury. PMID- 16355451 TI - Effects of the extract of a Chinese herb Tripterygium wilfordii hook f on rat pituitary gland. AB - In China, the ethylacetate extract of the herb Tripterygium wilfordii Hook f (TWEE), containing the major active ingredient triptolide, is often used with favorable effect on rheumatoid arthritis patients, in alternation with the use of prednisone. The mechanism of this therapeutic effect, however, has not been completely delineated. In this study, we studied how TWEE and prednisone affect the pituitary and adrenal glands in rats. Thirty normal male Sprague-Dawley rats (ten per group) were randomly assigned to receive: (1) TWEE (25 mg/kg, twice a day), (2) prednisone (2 mg/kg, twice a day), or (3) vehicle (control) (0.5% sodium carboxymethyl cellulose 1 ml, twice a day), orally for 30 days. Pituitary and trunk blood were collected on day 31. Adrenocorticotropic hormone (ACTH) expression in the pituitary gland was assessed morphologically by immunohistochemical techniques. Plasma ACTH concentrations and serum corticosterone concentrations were quantitatively measured by radioimmunoassay. We found that TWEE significantly increased plasma ACTH concentration and serum corticosterone concentration and dramatically increased the number of ACTH positive cells in the pituitary. Our findings indicate that TWEE can promote the synthesis and secretion of ACTH cells--in the pars distalis of the rat pituitary gland and the production of corticosterone in the zone fasciculata of the adrenal cortex. Our results indicate that TWEE has a cortical hormone-like function and can promote adrenal cortex function by activating the hypothalamus-pituitary adrenal axis. PMID- 16355452 TI - Inhibitory effects of Selaginella tamariscina on immediate allergic reactions. AB - The anti-allergic effects of a 70% ethanol extract from Selaginella tamariscina herb (EST) were evaluated in this study. EST given at the doses of 500 and 1000 mg/kg can inhibit mouse systemic anaphylactic shock induced by compound 48/80 in a dose-dependent manner. It can also dose-dependently block rat homologous passive cutaneous anaphylaxis and skin reactions caused by exogenous histamine and serotonin with a significant difference observed at the dose of 1000 mg/kg. In addition, EST can reduce histamine release from rat peritoneal mast cells triggered by compound 48/80 and an antigen in vitro. When incubated with rat mast cells, the extract (200 microg/ml) can significantly elevate the intracellular cAMP levels. The finding suggests that EST inhibits mast cell-dependent, immediate allergic reactions. Its effects appear to be mediated by reducing the release of vasoactive amines such as histamine from mast cells via stabilizing the cell membrane and weakening the inflammatory action of these amines. Based on these results, Selaginella tamariscina and one of its active components flavonoids may be useful as potential remedies for allergic rhinitis and other allergy-related diseases. PMID- 16355453 TI - Electropharmacological actions of the constituents of Sinomeni Caulis et Rhizome and Mokuboi-to in guinea pig heart. AB - Cardiac actions of the constituents of Sinomeni Caulis et Rhizome (SCR) and Mokuboi-to, a traditional herbal drug, were investigated. In voltage-clamp experiments, sinomenine (1 mM) and tetrandrine (100 microM) inhibited the ionic currents concentration-dependently. The constituents affected the action potential configurations. In multicellular preparations, SCR (1 mg/ml) decreased the maximum rate of depolarization. Tetrandrine (30 microM) and sinomenine (300 microM) also had similar effects, but magnoflorine (1 mM) had less or no effect. Dysrhythmias were abolished under Ca(2+) overload conditions by sinomenine. These results indicate that at even acute administrations, these drugs exert the active electropharmacological actions and cardioprotections. PMID- 16355454 TI - Antioxidative effects of tetramethylpyrazine on acute ethanol-induced lipid peroxidation. AB - Acute p.o. administration of 99.5% ethanol (0.1 ml) to fasted mice produced heart toxicity. Pretreatment with p.o. administration of tetramethylpyrazine (TMP) could prevent such toxicity effectively and dose-dependently. The maximal antioxidative effect against 99.5% ethanol-induced heart toxicity could be observed at 1 hour after TMP administration. In order to further investigate the heart protective mechanism of TMP, both lipid peroxidation level in vivo and superoxide scavenging activity were conducted. TMP exhibited a dose-dependently anti-lipid peroxidation effect in mice heart homogenate, and results indicated that 99.5% ethanol-induced intoxicated mice hearts have higher malonic dialdehyde (MDA) levels compared with those in TMP administrated mice hearts. These results suggest that the potentially heart protective mechanism of TMP could be contributed, at least in part, to its prominent anti-lipid peroxidation and anti free radical formation effects, hence it could protect the heart from lipid peroxidation-induced heart toxicity. PMID- 16355455 TI - Heart disease: opening the supply lines. PMID- 16355456 TI - Imaging: let's take a peek inside. PMID- 16355457 TI - Infectious diseases: reports of a demise were greatly exaggerated. PMID- 16355459 TI - Alzheimer's disease: scientific advances. PMID- 16355458 TI - Ophthalmology: seeing clearly now. PMID- 16355460 TI - A look ahead: plans for our November 2035 issue. PMID- 16355462 TI - Endocrine-related resources from the National Institutes of Health. PMID- 16355461 TI - Authors' response: HPA axis testing after pituitary surgery. PMID- 16355463 TI - Hormones & you. Patient information page. Erectile dysfunction. PMID- 16355465 TI - Medical dreams in Graeco-Roman times. PMID- 16355466 TI - For parents. Problem-solving school issues. PMID- 16355467 TI - Three clucks for poultry! PMID- 16355468 TI - American Diabetes Month. One size does not fit all. PMID- 16355469 TI - Tackling "diabesity" head-on. Joslin Diabetes Center's new nutrition guideline. PMID- 16355470 TI - Take a bite out of hypoglycemia. 10 proven strategies for cutting down on low blood glucose. PMID- 16355471 TI - Diabetes basics. Extreme makeover--recipe edition. PMID- 16355472 TI - Supermarket smarts. Chips, pretzels, and popcorn. PMID- 16355473 TI - Recipe makeovers. Grandma's Thanksgiving sausage stuffing. PMID- 16355474 TI - Women & diabetes. Self-management. Taming shaming and blame. PMID- 16355475 TI - Organ donation and transplantation. Understanding the basics. PMID- 16355476 TI - Pump training. Getting off to a good start. PMID- 16355477 TI - Diabetes quiz. How much do you know about potassium? PMID- 16355478 TI - Diabetes resources. Logbooks and record-keeping tools. PMID- 16355479 TI - An exploratory investigation of genetic linkage with body composition and fatness phenotypes: the Quebec Family Study. AB - In the present investigation, we have attempted to identify regions of the genome in which "obesity genes" potentially reside using robust sib-pair linkage analysis. Data were collected on 1,628 individuals in 301 nuclear families residing in the environs of Quebec City during the period 1978-1981. In addition to traditional blood group antigens and enzyme polymorphisms, several phenotypes in the obesity domain that are associated with increased morbidity were assessed, including measures relating to heaviness (i.e., the body mass index), body composition and nutrient partitioning (i.e., % body fat), and regional fat distribution without and with standardization for total fat mass (i.e., the sum of six skinfold thicknesses, and the ratio of the sums of trunk to extremity skinfold thicknesses). Three consistent patterns of potential linkage relationships with obesity phenotypes were revealed in these data, involving the marker loci adenosine deaminase, the Kell blood group antigen, and esterase D, which identify chromosomal regions 20q13, 7q33, and 13q14, respectively. Other potential linkages also were identified in the short arm of chromosome 1, interesting because of the presence of the db and fa loci on homologous regions of chromosome 1 in mouse and rat models of obesity, respectively. Each of the tentative linkage relationships reported here warrant follow-up using alternative methods and require replication in independent studies. PMID- 16355480 TI - Plasticity of brain alpha-adrenoceptors during the development of diet-induced obesity in the rat. AB - Male Sprague-Dawley rats, which are prone to develop diet-induced obesity (DIO) on a high energy (HE) diet can be separated from rats which are diet-resistant (DR) by several prospective tests. Using such tests, chow-fed DRl-prone rats have higher binding of 3H paraminoclonidine (PAC) to brain alpha2-adrenoceptors than do DIO-prone rats. These differences disappear after 3 months on a HE diet. To study the predictive value of these tests and possible associated changes in presynaptic membrane composition, brain alpha3(1-) (1nM 3H prazosin) and (alpha2 adrenoceptor (1nM) 3-H PAC) binding and synaptosomal fatty acid composition were assessed in 3-month-old male rats separated by weight gain into DR and DIO groups after 1 month on a HE diet. DIO had comparable total caloric intake but gained 30% and 43% more weight and were hyperinsulinemic compared to DR and chow-fed rats, respectively. After 1 month on a HE diet, DR rats still had 15%-53% higher 3H PAC binding than DIO and/or chow-fed rats in 14 of 16 brain areas assessed. A phenotype effect was present primarily in the amygdala where DR rats had higher 3H PAC binding than DIO rats. A diet effect was seen in some hypothalamic nuclei where both DR and DIO rats had higher 3H PAC binding than chow-fed rats. Conversely, DIO rats had 14%-21% higher 3H prazosin binding than DR rats in 3 brain areas. Changes in brain synaptosomal membranes' fatty acids reflected both phenotype and diet effects. Thus, while diet composition affects presynaptic membrane composition and alpha2-adrenoceptor binding in both DR and DIO rats, the predominance of plasticity of these parameters is limited to the brains of DR rats. This suggests that such plasticity may be an important determinant of the ability to resist the development of diet-induced obesity on a HE diet. PMID- 16355481 TI - The angle of the articular eminence in modern dentate African-Americans and European-Americans. AB - The purpose of this study was to determine if there were differences in the angle of eminentia of two 20th century populations based on race, age, gender, and number of teeth and whether there was asymmetry of the angles of eminentia. The sample included dry skulls from the Hamann-Todd Osteological collection as follows: 80 African-Americans (AA, 53 males and 27 females) and 62 European Americans (EA, 49 males and 13 females), ranging in age from 16-77 years. The lateral, central, and medial aspects of the right and left slopes of the articular eminence were measured in a parasagittal plane. Independent t-tests, paired t-tests, and Pearson correlation coefficients were computed. For the AA population, the right central, lateral, and medial angles of eminentia were steeper than the corresponding left angles (paired t-test, p<0.05); for the AE males only the right lateral and medial angles were significantly steeper than the corresponding left angles (paired t-test, p<0.05). There were no significant relationships between age or number of teeth and the angle of eminentia measurements, nor were there differences in angle of eminentia by gender. There were two differences by race: the EA males had steeper left central and left medial angles than the AA males (independent t-test, p<0.05). The central angle of eminentia was consistently steeper than the medial angle (paired t-test, p<0.01), and the lateral was generally steeper than the medial. PMID- 16355482 TI - Characteristics of the obesity syndrome in Zucker-Brown Norway (ZBN) hybrid rats. AB - The existence of a restriction fragment length polymorphism (RFLP) closely linked to the fatty locus between the Zucker (Z) and Brown Norway (BN) rat strains allows evaluation of early effects of the fatty (fa) gene using offspring of back crosses (N2) between F1 females and Zucker obese males. We examined several metabolic characteristics of N2 animals to determine if these hybrid animals exhibited similar characteristics of the obese syndrome to those of Zucker rats. Females from crosses of obese male Zucker (fa/fa) and lean female BN (+/+) rats were back-crossed to their sires, resulting in twelve N2 litters. At 9 weeks of age, liver, spleen, interscapular brown fat (IBAT), and gonadal, retroperitoneal (RP), and inguinal fat depots were removed and weighed. Samples of the RP depot were analyzed for cell size and number. Obese N2 rats were hyperphagic, with body weights in the range of those of obese Zucker rats. Obese N2 rats were also hyperinsulinemic [mean +/- SEM, microU/ml: females, 7.9 +/- 0.6 vs. 82.1 +/- 8.4 (lean vs. obese); males, 10.5 +/- 1.6 vs. 128.5 +/- 13.4 (lean vs. obese)] and mildly hyperglycemic [mean +/- SEM, mg/dl: females, 104.1 +/- 2.0 vs. 139.0 +/- 14.7 (lean vs. obese); males, 100.9 +/- 2.6 vs. 132.0 +/- 2.8 (lean vs. obese) p < or = 0.05]. White fat depots in obese rats were 3 to 7 times heavier than those in lean rats; adipocyte numbers in RP depots were 50% greater in obese than in lean rats; and cell size was more than 3 times larger. IBAT, liver, and spleen were also heavier in obese vs. lean rats, while tail lengths were shorter. Percent lean carcass mass and % carcass protein were about 30% greater in lean vs. obese rats, while % carcass fat in obese rats was 5 times greater than that of lean rats. Thus, phenotypic expression of the fa gene in ZBN hybrid animals, with approximately 25% of their genetic background coming from the BN strain, appears to be similar to that in Zucker rats. Given the similarity of phenotypic expression of the fa gene between the Zucker strain and ZBN hybrids, it is plausible to consider using ZBN hybrids for studies of early manifestations of fa gene action prior to onset of detectable obesity. PMID- 16355483 TI - Hypothalamic monoaminergic activity in 11-week-old cold-exposed female lean (Fa/Fa) and obese (fa/fa) Zucker rats. AB - We previously reported that serotonergic activity was reduced in the ventromedial hypothalamic nucleus (VMN) of obese vs. lean male Zucker rats. To verify that this reduction was associated with genotype rather than gender, we measured monoamines and their major metabolites in hypothalamic nuclei of 11-week-old female lean (Fa/Fa) and obese (fa/fa) Zucker rats. In addition, since the thermic response to cold is reported to differ between lean and obese rats, some rats were also exposed to 9 degrees or 22 degrees C for 2h to determine if cold exposure altered hypothalamic monoaminergic activity. As in males, levels of 5 hydroxyindoleacetic acid [5-HIAA; major metabolite of serotonin (5-HT)] and the ratio of 5-HIAA/5-HT were lower in the VMN of obese vs. lean females (P = 0.008, 0.001, respectively). 5-HIAA/5-HT was also reduced in the paraventricular (PVN) and suprachiasmatic nuclei (SCN) of the obese compared to the lean females. Cold exposure significantly stimulated brown fat mitochondrial GDP binding in lean but not obese rats. Similarly, levels of norepinephrine, dopamine (DA), 5-HIAA, and 5 HT in the PVN, and 5-HIAA in the SCN increased in cold-exposed lean but not obese rats. In contrast, VMN and preoptic 3,4-dihydroxyphenylacetic acid (DOPAC; major metabolite of DA) increased in the cold-exposed obese but not lean animals. We conclude that: (1) the blunted peripheral response to cold in obese vs. lean Zucker rats is accompanied by altered hypothalamic monoaminergic activity, the physiological role of which needs further evaluation; and 2) depressed VMN serotonergic activity is associated with the obese genotype (fa/fa) rather than gender and as such may contribute to the reduced sympathetic and enhanced parasympathetic outflow from the VMN. PMID- 16355484 TI - Extracellular hypothalamic monoamines measured by in vivo microdialysis in a rat model of dietary fat-induced obesity. AB - We tested two hypotheses about monoamine neurotransmitters in two strains of rats that differ in their sensitivity to obesity when eating a high-fat diet; 1) that the concentrations of norepinephrine and serotonin and of their metabolites differ in the extracellular fluid of the ventromedial hypothalamus of conscious, unrestrained Osborne-Mendel and S 5B/Pl rats, and 2) that these monoamines are altered differently between strains by a high-fat diet. The monoamines were measured by HPLC in dialysate collected by in vivo microdialysis in rats eating a semisynthetic low-fat diet (10% of kcal as fat) and again after either two or seven days of eating a high-fat diet (56% of kcal as fat). Norepinephrine, serotonin (5-HT), and 5-hydroxyindole-3-acetic acid (5-HIAA) were lower in Osborne-Mendel rats than in S 5B/Pl rats eating the low-fat diet. Norepinephrine and serotonin both increased in Osborne-Mendel rats with the onset of the high fat diet so that Osborne-Mendel and S 5B/Pl rats no longer differed in these neurotransmitters. By day 7 of high-fat feeding, the concentrations of 3-methoxy 4-hydroxyphenylglycol (MHPG), 5-HIAA and the 5-HIAA/5-HT ratio rose in both strains. Ambient extracellular monoamines in the medial hypothalamus are lower in Osborne-Mendel rats than in S 5B/Pl rats and the response of these catecholamines to dietary fat was greater in Osborne-Mendel rats than in S 5B/Pl rats. PMID- 16355485 TI - Positive modulation of the adipoconversion of adipoblasts derived from genetically obese rats by sodium butyrate. AB - Using a new serum-free primary culture system, we have previously reported genotypic differences between adipoblasts derived from the epididymal adipose deposit of lean and obese 8-week-old Zucker and Wistar Diabetic Fatty (WDF) rats (15). In these strictly controlled culture conditions, obese-derived adipoblasts expressed low levels of the late markers of differentiation (lipid accumulation, GPDH). In order to further characterize obese-derived adipoblasts and analyze the critical relationship between growth and differentiation, growth arrest was induced in lean- and obese-derived cultures using sodium butyrate treatment. Addition of 2.5 mM sodium butyrate to the serum-free medium from day 1 reduced markedly the growth of lean as well as obese-derived cells. Adipoconversion of lean-derived adipoblasts was not altered, similar levels of LPL and GPDH activities being obtained in control and butyrate-treated groups. By contrast, a marked increase in both activities was observed in obese-derived cultures, restoring the level of both markers of differentiation to the lean level. Similar results were obtained with adipoblasts derived from subcutaneous inguinal (ING) fat pad of obese Zucker as well as adipoblasts derived from ING and EPI fat deposits from obese WDF rats. Taken together, these results suggest that adipose deposits of these genetically obese rats contain a specific adipoblast population which differs from lean-derived adipoblasts in respect to its adipoconversion capacity and/or its stage of commitment to differentiation. PMID- 16355486 TI - Dieting status and its relationship to weight, dietary intake, and physical activity changes over two years in a working population. AB - The present study prospectively examined changes in dietary intake, physical activity and weight associated with self-reported efforts to lose weight in a cohort of 3671 men and women sampled from the general population. Dieting efforts, dietary intake, physical activity and weight were measured at two points in time, 24 months apart. At baseline, current dieters reported consuming fewer dairy products, sweets, meat, soft drinks and fried potatoes (all p's < .0001), and engaging more frequently in high-intensity physical activity (p < .0001) than those not currently dieting. At follow-up, current dieters reported consuming fewer sweets (p < .0001) and fried potatoes (p < .0008), and engaging more frequently in moderate-intensity physical activity (p < .02) than those not currently dieting. Prospectively, those who initiated weight-loss diets showed the largest decrease in consumption of sweets (p < .0001), soft drinks (p < .0001), and fried potatoes (p < .01), and increase in frequency of high-intensity physical activity (p < .0001) and moderate-intensity physical activity (p < .007). Those initiating weight-loss diets were the only group to lose weight (1 lb.). Those dieting at baseline but not at follow-up gained the most weight (4 lbs.). Self-reports of current dieting correspond to reported changes in dietary intake and physical activity, and to measured changes in weight over the same time period. Individuals who report dieting to lose weight have healthier eating and exercise patterns than those who do not report dieting. PMID- 16355487 TI - Obesity is bad for the heart, but is weight loss always good? PMID- 16355488 TI - Predictors of endoscopy in minority women. AB - BACKGROUND: Colorectal cancer (CRC) is the second leading cause of cancer related deaths in the United States. Underrepresented minorities suffer disproportionately from CRC largely because of disparities in CRC screening rates, particularly by endoscopic methods. This study evaluates the association between socioeconomic, medical and psychosocial factors and the use of endoscopy in low-income minority women. METHODS: The participants were recruited from community health fairs, tenant association meetings, senior centers and local medical clinics. A survey instrument was administered to the minority women. RESULTS: Eighty-one women age >50 were included in this analysis (44 African Americans and 37 Hispanics). The two ethnic groups were demographically similar. The factors associated with having had endoscopy were language spoken (English versus Spanish), physician recommendation, cancer cons and decisional balance (difference between cancer cons and cancer pros). When endoscopy was modeled as a function of decisional balance and language spoken, only decisional balance was a significant predictor of endoscopy. CONCLUSIONS: Physician recommendation and decisional balance have a tremendous influence on whether minority women undergo endoscopy. These data suggest that if physicians increase their communication with patients regarding the benefits of screening and address patients concerns, adherence with endoscopic CRC screening can be improved in minority women. PMID- 16355489 TI - Results of a faith-based weight loss intervention for black women. AB - Obesity is a risk factor for a variety of chronic diseases. Although weight loss may reduce these risks, weight loss programs designed for black women have yielded mixed results. Studies suggest that religion/spirituality is a prominent component of black culture. Given this, the inclusion of religion/spirituality as an active component of a weight loss program may enhance the benefits of the program. The role of religion/spirituality, however, has not been specifically tested as a mechanism that enhances the weight loss process. This paper presents the results of "Faith on the Move," a randomized pilot study of a faith-based weight loss program for black women. The goals of the study were to estimate the effects of a 12-week culturally tailored, faith-based weight loss intervention on weight loss, dietary fat consumption and physical activity. The culturally tailored, faith-based weight loss intervention was compared to a culturally tailored weight loss intervention with no active faith component. Fifty-nine overweight/obese black women were randomized to one of the two interventions. Although the results were not statistically significant, the effect size suggests that the addition of the faith component improved results. These promising preliminary results will need to be tested in an adequately powered trial. PMID- 16355490 TI - Sexually dimorphic expression of co-repressor Sin3A in mouse kidneys. AB - Using Western blot analysis we found transcriptional co-repressor Sin3A to be expressed at a higher level in male mouse kidney than in females. HDAC1 (histone deacetylase 1) protein, another co-repressor forming complexes with Sin3A, was not higher in males. No sex differences in Sin3A expression were found after gonadectomy, suggesting that gonadal secretions in adulthood cause the sex difference in kidney expression of Sin3A. In contrast, HDAC1 levels were higher in castrated gonadal males than in females, which presumably reflects a long lasting differentiating effect of testicular secretions in early development on this protein in kidneys. In gonadectomized mice in which sex chromosome complement (XX vs. XY) is independent of gonadal type (testes vs. ovaries), there was no difference in the level of Sin3A or HDAC1 expression in kidney in XX or XY mice of the same gonadal sex. PMID- 16355491 TI - Estrogen replacement raises rat CRP without evidence of complement activation. AB - Given current controversies regarding anti- and pro-inflammatory effects of estrogen, there is a need to explore relationships between gonadal hormones and inflammation using appropriate animal models. It has been proposed that rats are not appropriate for such research since, contrary to the effect of estrogen in humans, earlier animal studies had reported that estrogen downregulates serum C reactive protein (rCRP) levels in the rat. With these considerations in mind, we re-examined the effects of estrogen withdrawal and replacement on CRP expression and complement activation in the rat. F-344 rats underwent bilateral ovariectomy or sham surgery at 9-10 months of age. Four months later, ovariectomized rats were treated with traditional high-dose 17beta-estradiol (Hi-E2) capsules, lower dose (Lo-E2) 17beta-estradiol capsules, or placebo capsules for 7 days prior to sacrifice. Levels of plasma rat C-reactive protein (rCRP) were significantly lower in ovariectomized vs. sham-operated animals (415.5 +/- 10.6 vs. 626.6 +/- 23.0 mg/L, p < 0.001). Estrogen replacement significantly raised rCRP levels in ovariectomized animals (690.0 +/- 28.0 mg/L in Lo-E2 and 735.5 +/- 35.8 mg/L in Hi-E2, respectively, p < 0.001). Plasma rCRP levels correlated significantly with both hepatic rCRP (r = 0.79, p < 0.001) and serum estradiol (r = 0.70, p < 0.001) levels. However, no significant differences were observed in indices of complement activation (C4b/c) or CRP-complement complex generation (rCRP-C3 complex). In the mature female rat, ovariectomy reduces and estrogen replacement raises rCRP. Effects of estrogen on plasma rCRP induction are mediated, at least in part, through hepatic mechanisms and do not appear to require or be associated with complement activation. PMID- 16355492 TI - Nursing home closures, changes in ownership, and competition. AB - This research examines the relationship of competition among nursing homes and the likelihood of their closure or change in ownership. The study uses nationally representative data from the 1992-1998 Online Survey, Certification, and Reporting system, and is supplemented with several other primary and secondary data sources. It is hypothesized that facilities located in more competitive environments will be more likely to close. Multinomial logistic regression analyses are employed to examine this hypothesis in a model containing organizational and aggregate resident characteristics, and market factors. The Herfindahl index is used as a measure of competition. The descriptive analysis shows that 621 nursing homes closed and 6,471 changed ownership from 1992 to 1998. The incidence rate of closures was .7% of facilities per year. The multivariate analysis shows that facilities located in more competitive environments were significantly more likely to close. PMID- 16355493 TI - Will you recommend the flu shot for infants and children? PMID- 16355494 TI - Medical ethics and therapeutic progress: the example of lung cancer. Hippocrates to the rescue! PMID- 16355495 TI - Proceedings of the 5th International Conference on Nutrition and Fitness, Athens, Greece, June 9-12, 2004. Part 1. PMID- 16355496 TI - Proceedings of the 5th International Conference on Nutrition and Fitness, Athens, Greece, June 9-12, 2004. Part 2. PMID- 16355497 TI - Prevention of Renal Disease in the Emerging World: Toward Global Health Equity. Proceedings of the Bellagio Conference, March 16-18, 2004, Italy. PMID- 16355498 TI - Expanding the role of the forensic consultant. PMID- 16355499 TI - RETRACTION: Antimicrobial peptides (AMPs): peptide structure and mode of action. PMID- 16355500 TI - [Abstracts of the 9th Conference on Alzheimer Disease, Kecskemet, Hungary, September 21-23, 2005]. PMID- 16355501 TI - Serum TRACP 5b is a useful marker for monitoring alendronate treatment: comparison with other markers of bone turnover. AB - We studied clinical performance of serum TRACP 5b and other bone turnover markers, including S-CTX, U-DPD, S-PINP, S-BALP, and S-OC, for monitoring alendronate treatment. TRACP 5b had higher clinical sensitivity, area under the ROC curve, and signal-to-noise ratio than the other markers. INTRODUCTION: The purpose of this study was to compare the clinical performance of serum TRACP 5b (S-TRACP5b) with that of other markers of bone turnover in the monitoring of alendronate treatment. MATERIALS AND METHODS: This double-blinded study included 148 healthy postmenopausal women that were randomly assigned into two groups: one receiving 5 mg alendronate daily (n=75) and the other receiving placebo (n=73) for 12 months. All individuals in both groups received calcium and vitamin D daily. The bone resorption markers S-TRACP5b, serum C-terminal cross-linked telopeptides of type I collagen (S-CTX), and total urinary deoxypyridinoline (U DPD), and the serum markers of bone formation procollagen I N-terminal propeptide (S-PINP), bone-specific alkaline phosphatase (S-BALP), and total osteocalcin (S OC) were assessed at baseline and at 3, 6, and 12 months after initiation of treatment. Lumbar spine BMD (LBMD) was measured at baseline and 12 months. RESULTS: Compared with the placebo group, LBMD increased, and all bone markers decreased significantly more in the alendronate group (p<0.001 for each parameter). The decrease of S-TRACP5b after first 3 months of alendronate treatment correlated significantly with the changes of all other markers except S OC, the best correlation being with S-CTX (r=0.60, p<0.0001). The changes of LBMD at 12 months only correlated significantly with the changes of S-TRACP5b (r= 0.32, p=0.005) and S-CTX (r=-0.24, p=0.037) at 3 months. Based on clinical sensitivity, receiver operating characteristic (ROC) curves, and signal-to-noise ratio, S-TRACP5b, S-CTX, and S-PINP were the best markers for monitoring alendronate treatment. Clinical sensitivity, area under the ROC curve, and signal to-noise ratio were higher for S-TRACP5b than for the other markers. CONCLUSION: These results show that S-TRACP5b, S-CTX, and S-PINP are useful markers for monitoring alendronate treatment. PMID- 16355502 TI - Bone morphogenetic protein 2 induces cyclo-oxygenase 2 in osteoblasts via a Cbfa1 binding site: role in effects of bone morphogenetic protein 2 in vitro and in vivo. 2002. PMID- 16355503 TI - Preoperative planning for revision total hip arthroplasty. PMID- 16355504 TI - Saving lives through global safe water. PMID- 16355505 TI - Melioidosis in tsunami survivors. PMID- 16355506 TI - Chytrid fungus in Europe. PMID- 16355507 TI - Vibrio metschnikovii pneumonia. PMID- 16355508 TI - Lassa fever, Nigeria, 2003 and 2004. PMID- 16355509 TI - Methicillin-resistant Staphylococcus aureus skin infections. PMID- 16355510 TI - Angiostrongyliasis, Mainland China. PMID- 16355511 TI - Methicillin-resistant Staphylococcus aureus necrotizing pneumonia. PMID- 16355512 TI - West Nile virus infection and conjunctival exposure. PMID- 16355513 TI - Of tidal waves and human frailty. PMID- 16355514 TI - Diabetic foot: a major medical, social and economic problem in patients with diabetes. PMID- 16355515 TI - Energy intake and physical activity in Pima Indians: comparison with energy expenditure measured by doubly-labeled water. AB - To test the validity of survey techniques for measuring diet and activity patterns of Pima Indians, sequential 24-hour recalls, a food frequency questionnaire (FFQ), and an activity questionnaire were compared to free-living energy expenditure. Total energy expenditure (TEE) measured by doubly labeled water was 13.27 +/- 2.95 MJ/d for the 12 males (mean +/- SD: 35 +/- 14 yr; 97 +/- 35 kg; 32 +/- 9% body fat) and 11.67 +/- 1.85 MJ/d for the 9 females (31 +/- 13 yr; 106 +/- 32 kg; 49 +/- 6% body fat). Energy intake assessed by 24-hour recall was 13.59 +/- 7.81 MJ/d for men and 9.29 +/- 2.77 MJ/d for women, compared to 12.84 + 2.85 and 9.40 + 2.61 MJ/d for men and women, respectively, by FFQ. Both dietary methods indicated significant underreporting by women when compared to TEE. Energy intake assessed by FFQ was significantly correlated with TEE (r=0.48, p=0.03). This was true with 24-hour recall energy intake only when data from two extremely large alcohol consumers were eliminated (r=0.64, p=0.03, N=19). Although a low level of activity was apparent, the activity questionnaire produced significant correlations with measurements of energy expenditure and therefore represents an important tool for examining the relationship between physical activity and diseases. PMID- 16355516 TI - Intra-abdominal adipose tissue values associated with risk of possessing elevated blood lipids and blood pressure. AB - Central fat distribution and more recently intra-abdominal adipose tissue (IAF) have been associated with elevated cardiovascular risk factors (CRF). Despite increased interest in use of LAF for screening for CRF, interpretation of risk found in a specific IAF is difficult since regressions for estimating CRF from IAF have not been published. The purpose of this paper is to report IAF values that are likely to be associated with elevated CRF. One hundred forty-six healthy male subjects 30-71 years were evaluated for IAF and subcutaneous fat (computed tomography scan at 4th lumbar vertebra), height, body weight, % fat, various anthropometric measures, blood cholesterol (CHOL), HDL cholesterol (HDL), systolic blood pressure (SBP), and diastolic blood pressure (DBP). Receiver Operating-Characteristic curves (ROC) were used to develop IAF cutpoints associated with elevation of at least one established CRF criteria (CHOL=200, HDL=<35, SBP=140, DBP=90). A sensitivity/(1- specificity) curve established the value of using IAF cutpoints for detecting elevated CRF. Likelihood ratios were used to identify optimal cutpoints. Two cutpoints were identified, 131 cm2 with a relatively high Lpos ratio and 71 cm2 with a relatively low Lneg. False positives associated with 131 cm2 were 14% for one or more elevated CRF. False negatives associated with 71 cm2 were 9% for one or more elevated CRF, 4% for two or more CRF, and 0% for three or more elevated CRF. This study clearly indicates that IAF above 131 cm2 is related to elevated CRF and IAF below 71 cm2 is associated with reduced cardiovascular risk. PMID- 16355517 TI - Obesity prevention initiative. PMID- 16355518 TI - Towards prevention of obesity: research directions. PMID- 16355519 TI - NIDDK role in obesity research. PMID- 16355520 TI - The inheritance of corpulence. PMID- 16355521 TI - Body-build and its inheritance. 1923. PMID- 16355522 TI - Hereditary biological research. 1927. PMID- 16355523 TI - Nationwide Interoperability. When standards are available at the application level, our health infrastructure will be a reality. PMID- 16355524 TI - An action-PACS market. There's no end in sight for demands and use of medical imaging technology. PMID- 16355525 TI - Expertise times two. Collaboration between IT and radiology raises both proficiency and understanding. PMID- 16355526 TI - A deal from the top of the PACS. Organizations have more power than they realize to negotiate with PACS vendors. PMID- 16355527 TI - The architecture of sharing. An HL7 Version 3 framework offers semantically interoperable healthcare information. PMID- 16355528 TI - A provincial proving ground. By steering a regional registry, Mike Barron helps pave the way for Canada's EHR. PMID- 16355529 TI - In and out of surgery. For now, niche vendors fill the needs. PMID- 16355530 TI - Upgrading the RIS. The challenges and rewards are many. PMID- 16355531 TI - From N-WASP to WAVE: key molecules for regulation of cortical actin organization. AB - We first isolated N-WASP as one of the proteins bound to Ash/Grb2 SH3 domain. This protein has a VCA region (verplorin-like, cofilin-like, acidic region) at the C-terminus, which binds to G-actin and Arp2/3 complex, and several functional domains at the N-terminus, such as WHD (WASP homology domain) and GBD/CRIB domain. N-WASP activates Arp2/3 complex-dependent actin polymerization through the VCA region, leading to filopodium formation. Next, we found WAVE1, WAVE2 and WAVE3. All these proteins have also VCA regions at C-terminal areas and induce membrane ruffle formation. To clarify the different roles of WAVE1 and WAVE2, we established WAVE1- and WAVE2-deficient mouse embryonic fibroblasts (MEFs), because these two WAVEs are expressed in MEF. When wild-type MEFs are stimulated randomly by PDGF, two types of ruffles, peripheral and dorsal, are formed. However, dorsal ruffle formation does not occurin WAVE1-deficient MEFs. In contrast, peripheral ruffle formation is diminished in WAVE2-deficient MEFs. On the other hand, in MEFs migrating towards a chemoattractant gradient, only peripheral ruffles (lamellipodia) are formed. In this migration, WAVE1-deficient MEFs still could form lamellipodia but WAVE2-deficient MEFs could not. All these data show that WAVE2 but not WAVE1 is essential for lamellipodium formation and directed migration. PMID- 16355532 TI - A conserved role for myosin VII in adhesion. AB - The class VII myosins (M7) are expressed in a wide range of organisms. M7 mutants in mice, zebrafish and Dictyostelium exhibit phenotypes that reveal a role for M7 in adhesion in these highly divergent systems, suggesting a basic conservation of M7 function throughout evolution. M7s are characterized by the presence of two FERM domains in their C-terminal tail region, and deletion of either from the Dictyostelium M7 (DdM7) tail results in loss of function without affecting localization. A search for DdM7 binding partners has revealed that talin, an actin-binding protein that provides a key link between adhesion receptors and the actin cytoskeleton, interacts directly with DdM7. The phenotypes of the DdM7 and talin null mutants are highly similar, suggesting that these two proteins work co operatively to maintain cell-cell and cell-surface contact and that this interaction may also be conserved throughout evolution. PMID- 16355533 TI - Cytoskeletal networks and pathways involved in endocytosis. AB - Until recently, the actin cytoskeleton and the endocytic machinery were thought to operate independently. However, the actin cytoskeleton is an integral part of the cell cortex and there is growing evidence in diverse eukaryotes that F-actin plays a direct role during endocytic internalization. Genetic studies in Saccharomyces cerevisiae have demonstrated that Arp2/3-mediated F-actin assembly is required specifically for the internalization step of endocytosis. Using real time image analysis, we recently defined a pathway for receptor-mediated endocytosis in budding yeast. Many features of this pathway appear to be conserved widely, indicating that principles derived from our studies in yeast will be directly applicable in more complex eukaryotes. We are pursuing our yeast studies using a combined approach involving image analysis, functional genomics, proteomics and biochemistry. These ongoing studies are providing a broader and deeper understanding of the molecular events of endocytosis, of how forces for actin polymerization are harnessed, and of how steps in the pathway are regulated. Our studies in mammalian cells provide evidence that this pathway is conserved in more complex organisms for endocytic and Golgi trafficking events. PMID- 16355534 TI - Control of cell polarity in response to intra- and extracellular signals in budding yeast. AB - Budding yeast serves as a powerful genetic model organism for studying the molecular mechanisms of cell polarity in single cells. Like other polarized eukaryotic cells, yeast cells possess polarity programs that regulate where they grow and divide. Establishment of a site of cell polarity may be conceptualized in several stages. First, cells mark a specific location at the cell surface for polarized cell growth and cell division. To define these sites, cells use intrinsic cues present in the cell or landmarks determined by extracellular signals such as morphogens. Second, these landmark proteins then recruit or activate polarity establishment proteins including small GTPases and their regulators. Positive and negative feedback mechanisms are required to transform these site-selection processes into a stable axis of polarity. Finally, these locally activated GTPase modules recruit and activate proteins that organize the actin cytoskeleton and cell growth. In this short review, we describe molecular pathways required to establish oriented cell polarity, and emphasize recent advances in defining positive and negative feedback mechanisms that together may translate an initially weak symmetry-breaking signal into a robust axis of polarity. PMID- 16355535 TI - Regulation of actin assembly by microtubules in fission yeast cell polarity. AB - It has been speculated that microtubule plus ends function to regulate the actin cytoskeleton in processes such as cytokinesis, cell polarization and cell migration. In the fission yeast Schizosaccharomyces pombe, interphase microtubules regulate cell polarity through proteins such as tea1p, a kelch repeat protein, and for3p, a formin that nucleates actin cable assembly at cell tips. Here, we review recent progress on understanding tea1p regulation and function. Microtubules may govern the localization of tea1p by transporting it on the plus ends of microtubules and depositing it directly onto the cell tip when the microtubule catastrophes. The interaction of tea1p with the CLIP170 protein tip1p is responsible for its localization at growing microtubule plus ends. Tea1p may regulate cell polarity by associating with large 'polarisome' complexes that include for3p. For3p is present at both cell tips, but is not on the microtubules. Tea1p is needed to localize the formin to establish polarized cell growth at cell tips that have not grown previously. These studies begin to elucidate a molecular pathway for how microtubules contribute to the proper spatial regulation of actin assembly and polarized cell growth. PMID- 16355536 TI - Finding the way: directional sensing and cell polarization through Ras signalling. AB - Chemotactic eukaryotic cells have the unique ability to sense a shallow extracellular chemoattractant gradient and translate it into a steep intracellular gradient. For example, phosphoinositide-3,4,5-trisphosphate (PIP3), the product of phosphatidylinositol-3-kinase (PI3K), is accumulated at the leading edge but not the back of a polarized chemotaxing cell. This is partially controlled by the reciprocal, preferential localization of PI3K and PTEN to the membrane at the front and back, respectively. However, upstream events that control the localized activation and localization of PI3K and PTEN remain unclear. Recent findings indicate that Ras is important for activation of the PI3K pathway and regulation of directed cell movement and cell polarity. Ras is activated at the leading edge, and this local activation occurs without asymmetric localization of PI3K and PTEN or the F-actin cytoskeleton. In contrast, P13K localization is driven by F-actin polymerization. Thus, Ras functions as an essential part of the cell's compass acting upstream of PI3K while reciprocal localization of PI3K and PTEN amplify the PIP3 gradient, rather than create it. These observations suggest a positive feedback loop to amplify an initial PIP3 gradient in which localized F-actin polymerization recruits cytosolic PI3K to the leading edge, where it is activated by Ras to locally produce PIP3 that induces F-actin polymerization. PMID- 16355537 TI - Roles of IQGAP1 in cell polarization and migration. AB - Cell polarization and migration are fundamental processes in all organisms and are stringently regulated during tissue development, chemotaxis and wound healing. Migrating cells have a polarized morphology with an asymmetric distribution of signalling molecules and the cytoskeleton. Linkage of microtubule plus ends to the cortical region is essential for polarized migration. +TIPs, including CLIP-170 and APC (adenomatous polyposis coli) are thought to function as capturing devices at specialized cortical regions. Rho family GTPases, particularly Rac1 and Cdc42, play pivotal roles in cell polarization and migration acting through their effectors. We found that IQGAP1, an effector of Rac1 and Cdc42, interacts with CLIP-170. Activated Rac1 and Cdc42 enhance the binding of IQGAP1 to CLIP-170, and capture GFP-CLIP-170 at the base of leading edges and filopodia, respectively. Recently, we found that IQGAP1 directly binds to APC in addition to CLIP-170. IQGAP1 and APC interdependently localize to leading edges in migrating cells. IQGAP1 can link APC to actin filaments in vitro. Thus, activation of Rac1 and Cdc42 in response to migration signals leads to recruitment of IQGAP1 and APC which, together with CLIP-170, form a complex that links the actin cytoskeleton and microtubule dynamics during cell polarization and migration. PMID- 16355538 TI - Actin organization in the early Drosophila embryo. AB - Organization of the cortical cytoplasm during the syncytial blastoderm stages of early Drosophila embryogenesis relies on cyclic transitions between transient microfilament structures. Microtubule-organizing centres (MTOCs) appear to provide the instructive cues governing this dynamic, cell-cycle-dependent process. Using a genetic approach, we have identified key roles for two molecular pathways in mediating these events. The conserved Arp2/3 microfilament nucleation machinery, likely acting in response to the activating element SCAR, plays an essential role in establishment of a cortical F-actin array, and contributes to specific aspects of cyclic microfilament restructuring. Defective cortical microfilament organization is the primary phenotypic feature of embryos derived from mothers bearing mutations in the sponge locus. Several lines of investigation suggest that the primary defect in sponge lies in a faulty cortical microfilament response, downstream of the centrosomal signal. We have determined that sponge encodes a Drosophila homologue of the evolutionarily-conserved CDM (DOCK180) protein family. PMID- 16355539 TI - Epithelial cell shape and Rho small GTPases. AB - In epithelial cells, morphology is intrinsically related to function. Typically, polarization (i.e. acquisition of a cuboidal cell shape) must occur prior to terminal differentiation and functionality of epithelial sheets. Extensive work has been performed to understand the initial steps that drive cell-cell contact assembly, a process that is essential for polarization. However, not much is known about the subsequent steps that lead to remodelling of actin cytoskeleton and concomitant cell shape changes. Here we review what is known about actin organization during epithelial polarization, and discuss current models for junction assembly, actin reorganization and signalling pathways that may contribute to the generation of a polarized epithelial morphology. PMID- 16355540 TI - Integrin-syndecan cooperation governs the assembly of signalling complexes during cell spreading. AB - Cell adhesion to fibronectin (FN) triggers the formation and maturation of adhesion complexes by modulating the activity of the Rho family of GTPases. Cells plated onto a ligand of integrin alpha5beta1 spread but fail to form focal adhesions or fully organize actin into bundled stress fibres unless co-stimulated with a ligand of syndecan 4. Engagement of syndecan 4 in such pre-spread cells recapitulates the Rac1 and RhoA activation profiles observed during spreading on whole FN. Furthermore, since adhesion to a ligand of alpha5beta1 alone does not activate Rac1, engagement of syndecan 4 appears to be an absolute requirement. In related work, we have examined differences in the mechanism of focal adhesion formation mediated by the FN-binding integrins alpha4beta1 and alpha5beta1. Two signalling differences were found. First, while alpha5beta1 required syndecan 4 as a co-receptor, alpha4beta1 did not. Second, focal adhesion formation via alpha5beta1 required PKCalpha activation, but only basal PKCalpha activity was observed following adhesion via alpha4beta1. These findings demonstrate that different integrins can signal to induce focal adhesion formation by different mechanisms. PMID- 16355541 TI - Formation of multicellular epithelial structures. AB - The kidney is primarily comprised of highly polarized epithelial cells. Much has been learned recently about the mechanisms of epithelial polarization. However, in most experimental systems the orientation of this polarity is determined by external cues, such as growth of epithelial cells on a filter support. When Madin Darby canine kidney (MDCK) cells are grown instead in a three-dimensional (3D) collagen gel, the cells form hollow cysts lined by a monolayer of epithelial cells, with their apical surfaces all facing the central lumen. We have found that expression of a dominant-negative (DN) form of the small GTPase Rac1 causes an inversion of epithelial polarity, such that the apical surface of the cells instead faces the periphery of the cyst. This indicates that the establishment of polarity and the orientation of polarity can be experimentally separated by growing cells in a 3D collagen gel, where there is no filter support to provide an external cue for orientation. DN Rac1 causes a defect in the assembly of laminin into its normal basement membrane network, and addition of a high concentration of exogenous laminin rescues the inversion of polarity caused by DN Rac1. PMID- 16355542 TI - An overview of methods for interval-censored data with an emphasis on applications in dentistry. AB - Interval-censored time-to-event data occur in many medical areas, with dentistry or AIDS research being typical representatives. This article reviews methods for the analysis of such data, with an emphasis on the use of the accelerated failure time (AFT) model. A flexible AFT model (avoiding parametric assumptions on the distribution of the error term) is described in greater detail and is used to solve a typical dental question in a longitudinal oral health study. PMID- 16355543 TI - A multilevel modelling solution to mathematical coupling. AB - Owing to mathematical coupling, statistical analyses relating change to baseline values using correlation or regression are erroneous, where the statistical procedure of testing the null hypothesis becomes invalid. Alternatives, such as Oldham's method and the variance ratio test, have been advocated, although these are limited in the presence of measurement errors with non-constant variance. Furthermore, such methods prohibit the consideration of additional covariates (e.g., treatment group within trials) or confounders (e.g., age and gender). This study illustrates the more sophisticated approach of multilevel modelling (MLM) which overcomes these limitations and provides a comprehensive solution to the analysis of change with respect to baseline values. Although mathematical coupling is widespread throughout applied research, one particular area where several studies have suggested a strong relationship between baseline disease severity and treatment effect is guided tissue regeneration (GTR) within dental research. For illustration, we use GTR studies where the original data were found to be available in the literature for reanalysis. We contrast the results from an MLM approach and Oldham's method with the standard (incorrect) approach that suffers from mathematical coupling. MLM provides a robust solution when relating change to baseline and is capable of simultaneously dealing with complex error structures and additional covariates and/or potential confounders. PMID- 16355544 TI - A Bayesian analysis of amalgam restorations in the Royal Air Force using the counting process approach with nested frailty effects. AB - Survival analysis methods are increasingly used in dental research to measure risk of tooth eruption and caries as well as life spans of amalgam restorations. Analyses have been extended to account for lack of independence in the data, which arises from the clustering of observations within units such as tooth surfaces, teeth and subjects. There are various analytical strategies and modelling approaches now available to us in dealing with clustered dental data. In this article, the modelling strategy of Cox's proportional hazards regression is formulated using the counting process approach, which can easily be extended to include time-variant covariates as well as nested random frailty effects. A semi-parametric Bayesian method is presented for the analysis of the proposed model. The methodology is applied to an analysis of nested clustered data on life span of amalgam restorations in the UK Royal Air Force. These data have previously been analysed using a non-Bayesian approach. The Gibbs sampler, a Markov chain Monte Carlo method, is used to generate samples from the marginal posterior distribution of the parameters of this Bayesian model. PMID- 16355545 TI - Endpoints and survival analysis for successful osseointegration of dental implants. AB - We review and adapt modern survival analysis adapted for evaluating the success of dental implants is reviewed. A defined composite success criterion is a valuable tool for assessing new developments of dental implants. However, several inaccuracies are detected in the relevant dental literature. The Kaplan-Meier estimator is distinguished from ad-hoc life table methods and crude cumulative percentages, and generally, the usage of confidence intervals is emphasized. This article reviews the definition of implant success also. For the example of longitudinal measurements of the peri-implant bone level, it is shown how to derive success probabilities by time-to-event analysis. It is concluded that advanced statistical methods and well-defined endpoints are needed to achieve meaningful and comparable results. The article is illustrated using the data of a familiar implant system (TPS-SteriOss). PMID- 16355546 TI - Group sequential testing in dental clinical trials with longitudinal data on multiple outcome variables. AB - In this article, methods are proposed for design and analysis of clinical trials that gather longitudinal data on multiple outcome variables. A valid test of the null hypothesis of no treatment group differences can be obtained for any choice of a working alternative hypothesis and a working covariance matrix for the outcome variables. Increased power can be achieved by accurate modeling of the true treatment effect and covariance structure. Implementation of the procedure is simple using existing software for generalized estimating equations. The procedure is an extension of the 'derived variable' technique (univariate analysis applied to a linear combination of the outcome variables) and also of O'Brien's generalized least squares test. The procedure is extended to allow sequential testing using an arbitrary division of the total type I error rate among repeated hypothesis tests. The methods are illustrated by the design of a study on the safety of dental amalgam fillings, which served as the motivation for the research. PMID- 16355547 TI - Treatment of anemia with darbepoetin alfa in the radiation oncology setting. PMID- 16355548 TI - Development of a strategy to successfully convert an ambulatory infusion center from epoetin alfa to darbepoetin alfa. PMID- 16355549 TI - Hemoglobin response and improvements in quality of life in anemic children with cancer receiving myelosuppressive chemotherapy. PMID- 16355550 TI - Advances in conducting retrospective chart review studies: use of electronic report forms and database in a community study in anemic patients with myelodysplastic syndrome. PMID- 16355551 TI - Optimizing chemotherapy-induced anemia management with the use of patient flow sheets. PMID- 16355552 TI - Iron, folate, and cancer-related anemia: perfect together. PMID- 16355553 TI - A multidisciplinary approach for establishing standardized anemia procedures. PMID- 16355554 TI - Utilizing quality of life as a factor in treatment choices for chemotherapy induced anemia. PMID- 16355555 TI - Admission and care of the febrile neutropenic patient in a community hospital setting. PMID- 16355556 TI - Predictors of chemotherapy-induced neutropenia and its complications: results from a prospective nationwide registry. PMID- 16355557 TI - Best practices in neutropenia management: you can get there from here. PMID- 16355558 TI - Identification and verification of symptom clusters in cancer patients. PMID- 16355559 TI - Advancing our knowledge of symptom clusters. PMID- 16355560 TI - Profile of cancer-related symptoms prior to chemotherapy. PMID- 16355561 TI - TITAN: modifying an innovative American program for use with a European audience. PMID- 16355562 TI - Baseline evaluation of assessment, information, and management patterns in a national quality improvement project. PMID- 16355563 TI - Cell transplantation: stem cells in the spotlight. PMID- 16355564 TI - Beta1 integrin as a xenoantigen in fetal porcine mesencephalic cells transplanted into the rat brain. AB - Xenografts of fetal porcine mesencephalic cells implanted into the rat striatum are generally rejected within several weeks. The fetal donor mesencephalon predominantly consists of neurons, but also contains microglial and endothelial cells, which are more immunogenic. In the present work, we investigated the occurrence of donor endothelial cells in grafts of porcine mesencephalic cells implanted into the rat striatum. Pig endothelial cells were monitored by immunochemical methods, using a monoclonal antibody (mAb) that recognizes a peptidic epitope of the porcine beta1 integrin, and isolectin IB4, for the staining of the Galalpha1,3Gal epitope. The analysis also involved the detection of the pig hyaluronate receptor CD44, and the cell adhesion molecule CD31. The anti-beta1 integrin mAb revealed endothelial-like cells in grafts of porcine mesencephalic cells as soon as 1 week after implantation. A similar staining pattern was obtained with the IB4 lectin. Unlike aortic endothelial cells, these pig brain-derived endothelial-like cells were not recognized by the anti-CD44 antibody. They also failed to express the CD31 adhesion molecule, a fact which suggests that they remained poorly mature, even in grafts maintained during 45 days in immunosuppressed rats. Interestingly, a strong expression of beta1 integrin immunoreactivity was noticed in a large proportion (80%) of the cells freshly dissociated from the fetal pig mesencephalic tissue. The immunoreactivity decreased progressively after transplantation of the cells into the rat brain. This observation suggests that dissociated neuroblasts are capable of a temporary expression of beta1 integrin. This molecule is known to participate in the process of cell sorting and migration in the developing brain. Hence, its expression could be the hallmark of a rescue mechanism triggered by the disruption of the cell/matrix interactions during the dissociation of the fetal mesencephalon. This disruption might account for part of the dramatic cell death process that occurs during the manipulation of the donor tissue. PMID- 16355565 TI - Creatine supplementation improves dopaminergic cell survival and protects against MPP+ toxicity in an organotypic tissue culture system. AB - Cell replacement therapy using mesencephalic precursor cells is an experimental approach for the treatment of Parkinson's disease (PD). A significant problem associated with this procedure is the poor survival of grafted neurons. Impaired energy metabolism is considered to contribute to neuronal cell death after transplantation. Creatine is a substrate for mitochondrial and cytosolic creatine kinases (CK) and buffers cellular ATP resources. Furthermore, elevated cellular creatine levels facilitate metabolic channeling and show antiapoptotic properties. Exogenous creatine supplementation therefore might offer a tool for improvement of dopaminergic neuron survival. The present study aimed at investigating the effects of creatine on cell survival of rat embryonic day 14 (E14) ventral mesencephalic neurons grown as organotypic free-floating roller tube (FFRT) cultures. We found that the brain-specific isoform of CK (BB-CK) and the ubiquitous mitochondrial isoform (uMt-CK) are expressed at high levels in FFRT cultures and colocalize with tyrosine hydroxylase immunoreactive (TH-ir) cells. Exposure of these cultures to creatine induced an increase in the content of the BB-CK isotype. Creatine (5 mM) administration starting at day in vitro (DIV) 7 resulted in a significant increase (+35%) in TH-ir cell density at DIV21. In addition, we observed that creatine treatment provided neuroprotection against 1-methyl-4-phenyl pyridinium ion (MPP+)-induced TH-ir cell loss in the FFRT culture system, resulting in a significantly higher density (+19%) of TH-ir neurons in creatine-treated cultures compared to corresponding controls. The decrease of TH-ir neurons in the MPP+-treated group corresponded with an increase in immunoreactivity for active caspase-3, an effect that was not seen in the group receiving creatine supplementation. In conclusion, our data imply that creatine administration is beneficial for the survival of TH-ir neurons encountering harmful conditions. PMID- 16355566 TI - Selection of cell source for ligament tissue engineering. AB - Use of appropriate types of cells could potentially improve the functionality and structure of tissue engineered constructs, but little is known about the optimal cell source for ligament tissue engineering. The object of this study was to determine the optimal cell source for anterior cruciate ligament (ACL) tissue engineering. Fibroblasts isolated from anterior cruciate ligament, medial collateral ligament (MCL), as well as bone marrow mesenchymal stem cells (MSC) were compared using the following parameters: proliferation rate, collagen excretion, expression of collagen type I, II, and III, as well as alpha-smooth muscle actin. Green fluorescent protein (GFP) transfected MSCs were used to trace their fate in the knee joints. MSC, ACL, and MCL fibroblasts were all highly stained with antibodies for collagen types I and III and alpha-smooth muscle actin while negatively stained with collagen type II. Proliferation rate and collagen excretion of MSCs were higher than ACL and MCL fibroblasts (p < 0.05), and MSCs could survive for at least 6 weeks in knee joints. In summary, MSC is potentially a better cell source than ACL and MCL fibroblasts for anterior cruciate ligament tissue engineering. PMID- 16355567 TI - The effects of cryopreservation on human hepatocytes obtained from different sources of liver tissue. AB - Successful cryopreservation of human hepatocytes is important to establish hepatocyte banks for clinical use or in vitro research. The availability of donor tissue from unused liver segments/lobes and non-heart-beating donors (NHBD) has provided newer sources of hepatocytes. The quality of hepatocytes at the time of cryopreservation is important as cells isolated from liver tissue of borderline quality may not withstand the stresses associated with cryopreservation and subsequent thawing. Human hepatocytes were cryopreserved after isolation from mainly donor tissues (n = 40). In vitro assessment of the viability and function of the fresh and thawed cryopreserved hepatocytes was performed. Viability, attachment efficiency, enzyme activity, and albumin production of hepatocytes were all significantly decreased, and LDH leakage significantly increased, on thawing after cryopreservation. The viability of cryopreserved hepatocytes isolated from tissue rejected for orthotopic liver transplantation (36 +/- 15%) was significantly lower than those isolated from tissue where part was used for liver transplantation (47 +/- 14%, p = 0.002), but there were no significant differences in functional parameters. The viability of cryopreserved hepatocytes isolated from NHBD tissue (29 +/- 9%, p = 0.001) and from steatotic donor tissue (35 +/- 11%, p = 0.019) was significantly lower than those isolated from normal donor tissue (49 +/- 14%). There was no difference in functional parameters, except for albumin production of hepatocytes from NHBD tissue (2.9 +/- 1.0 microg/h/mg protein) being significantly lower than those from normal donor tissue (4.8 +/- 2.8 microg/h/mg protein, p = 0.03). The viability and attachment efficiency of cryopreserved hepatocytes isolated from liver tissue from resections for tumors was significantly higher, and the LDH leakage significantly lower, than those isolated from all donor tissue. Hepatocytes isolated from NHBD and steatotic tissue were more vulnerable to the effects of cryopreservation. Further research is required to improve hepatocyte isolation and cryopreservation protocols for different types of liver tissue. PMID- 16355568 TI - Subcutaneous transplantation of islets into streptozocin-induced diabetic rats. AB - Pancreatic islet transplantation into type 1 diabetic patients is currently being performed by intraportal infusion. This method, albeit reproducible, has some disadvantages including potential development of portal hypertension, hemorrhage, and an inability to retrieve or detect the transplanted tissue. Other transplant sites have been examined in animal models including the omentum, peritoneal cavity, and the spleen. A transplant site that has not been successful in supporting functional islet tissue transplantation in humans is the subcutaneous space due primarily to the lack of a well-defined vascular bed. This site has many favorable characteristics such as ease of access for transplantation and potential for removal of the transplanted tissue with a minimally invasive surgical procedure. This report addresses the evaluation of a subcutaneously placed device for the support of rat syngeneic islet transplantation in a streptozocin-induced diabetic model. The data generated support the use of this device for islet engraftment. In addition, beta cell function in this device compared favorably with the function of islets transplanted to the renal subcapsular space as well as islets within the native pancreas. PMID- 16355569 TI - [Complications of an unrecognized myocardial infarction]. AB - Patients with an untreated myocardial infarction may present with serious late complications. 3 patients are described. A 63-year-old woman became progressively more short of breath 4 days after an acute episode of chest pain accompanied by nausea and sweating. It proved to be a cardiogenic shock following a rupture of a papillary muscle. A man aged 65 collapsed 5 days after an episode of back pain and nausea. This was a cardiac tamponade due to rupture of the left ventricle. A woman aged 74 had transient aphasia and during investigations for this was seen to have anomalies on ECG. She had cerebral emboli and a cardiac aneurysm with associated thrombus. All 3 patients recovered following mitral-valve replacement, repair of the rupture and medicinal treatment for the clot, respectively. Around one-third of patients who have a myocardial infarction do not have chest pain but experience shortness of breath, autonomic nervous symptoms (sweating, nausea, vomiting), extreme and inexplicable tiredness and fainting. These atypical symptoms should suggest myocardial infarction. In order to avoid high morbidity and death from complications such as arrhythmias, heart failure, rupture and aneurysm formation it is important that a patient who has had a myocardial infarction should be treated as soon as possible, preferably by reperfusion therapy. PMID- 16355570 TI - [Meningococci infection by a bacteriophage with a virulence factor]. AB - Meningococci are bacteria dreaded for their ability to kill young people. However, meningococci and humans usually live together peacefully. In a minority of cases, the co-existence results in disease. Recently, whole genome comparisons between hyperinvasive clones and clones not associated with disease revealed that a chromosomally integrated bacteriophage was related to invasiveness. Many examples of bacteriophage-encoded virulence factors are known--as such, this finding is not remarkable. However, the way this virulence factor was found is a nice example of unravelling the pathogenesis of infectious diseases in the genomic era. PMID- 16355571 TI - [Prevention of female genital mutilation in The Netherlands]. AB - Political attention was drawn to the item of female genital mutilation in The Netherlands in 2004. A task force has been installed to investigate the incidence of and evaluate the attitude towards female genital mutilation. The incidence is estimated to be 50 girls a year, especially girls from Africa. The government has decided to draw up rules for physicians and nurses in paediatric public health in order to influence the attitude of women and help them to abandon this tradition. PMID- 16355572 TI - [Lifestyle intervention for the prevention of cardiovascular disease]. AB - The high cardiovascular disease prevalence in western countries is largely attributable to the contemporary lifestyle. Interventions in the area of nutrition and physical activity have been shown to be effective in the prevention of cardiovascular disease. Successful implementation of lifestyle intervention programmes may be just as effective as drug treatment. In combination with drug treatment, intervention in the area of nutrition and physical activity is the recommended treatment for patients at a high risk of cardiovascular disease. Addition of new drugs to those presently available is associated with low absolute risk reductions and high costs, particularly in the presence of successful lifestyle interventions. PMID- 16355573 TI - [The practice guideline 'Blood transfusion' (third integral revision)]. AB - The revised and expanded practice guideline 'Blood transfusion' describes the whole transfusion chain within the hospital for the first time. Despite compatibility tests before transfusion (determination of the ABO and Rhesus blood groups and detection of clinically relevant antibodies (C, c, D, E, e, Fy(a), Fy(b), Jk(a), Jk(b), M, S and s)), transfusion reactions can occur. So that a transfusion reaction can be recognised in time, the patient must be observed intensively for the first 5-10 minutes after the start of any new transfusion and the vital functions must be recorded. In patients with a Hb level of 4-6 mmol/l, the decision whether or not to transfuse should be made dependent on the patient's other characteristics. Thrombocyte transfusion is not indicated in case of thrombopenia due to increased breakdown or pooling. If leukaemia, tumour infiltration or drug toxicity is the underlying cause of thrombopenia, then a platelet count of 10 x 10(9)/l or 20 x 10(9)/l should be the transfusion trigger. Reduction of the number of blood transfusions can be achieved by the administration of epoetin in case of renal insufficiency: transfusion can thus be avoided in more than 70% of the patients concerned. Autotransfusion during surgery with severe blood loss also results in a reduction of the number of allogenic blood transfusions. PMID- 16355574 TI - [From gene to disease; a progressive cochlear-vestibular dysfunction with onset in middle-age (DFNA9)]. AB - DFNA9 is an autosomal dominant genetic inner-ear hearing impairment that starts to show itself in the 3rd and 4th decades of life. This hearing impairment may be of a different degree of severity in each ear. Progression of hearing loss is about 3 dB/year. In about one in three patients severe vestibular symptoms similar to those in Meniere's disease are present as a result of a progressive impairment of the vestibular system. Several mutations were found in the COCH gene on chromosome 14. There are indications that some of the mutations disrupt the folding of the cochlin protein, an important component of the extracellular matrix in the inner ear. DNA-diagnostics confirming the diagnosis ofDFNA9 are possible. PMID- 16355575 TI - [Transfusion reactions in patients: haemovigilance reports to the Dutch National Haemovigilance Office in 2003]. AB - OBJECTIVE: National registration and analysis of unexpected side effects and incidents associated with blood transfusion (together termed 'transfusion reactions') in 2003 in order to arrive at recommendations to improve safety in the transfusion chain. DESIGN: Observational METHOD: A uniform national reporting form with definitions and a reporting manual were sent to all Dutch hospitals in the spring of 2003 with the request to report transfusion reactions retroactively to January 2003 to the Dutch National Haemovigilance Office 'Transfusion reactions in patients' (TRIP). TRIP is an independent organisation managed by representatives of professional societies that are involved in blood transfusion. Each hospital was given a reporting code. The reports were in principle voluntary and anonymous with regard to both the patient and the attending physician. Transfusion reactions were assessed for severity as well as for the level of probability with which they could be ascribed to the transfusion. RESULTS: Reports were received from 82 (80%) of the hospitals; 9 hospitals informed the TRIP explicitly that there had been no transfusion reactions in 2003. A total of 267 reports were received. Of these, 803 (63%) were graded for severity and of these 803, 52 (6%) were grade 2 ('moderate to severe morbidity') or worse. In the categories involving possible infectious complications, there were 2 reports of bacterial contamination that were judged, on review, to be due 'with certainty' to the transfusion. 34 reports concerned transfusion of the wrong blood product, resulting in a total of 9 transfusion reactions (4 of grade 2). The total number of reports concerning 2003 was 1.6/1000 blood products. CONCLUSION: The participation by the hospitals was high in the first year of national reporting, 2003. Most of the reports were of non-serious reactions known to be possible side effects of blood transfusion; 52 reports were rated as grade 2 or worse. PMID- 16355576 TI - [Severe bleeding in a patient with anti-c alloantibodies and a rare Rhesus phenotype treated with compatible erythrocyte concentrate from the blood bank of the Council of Europe]. AB - An 84-year-old women had repeated gastrointestinal bleeding from a Dieulafoy lesion, i.e. a gastric or duodenal ulcer containing an aberrant artery. Her blood group was AB-D negative; her Rhesus phenotype was CCdee. In addition, antibody screening revealed anti-c alloantibodies as the result of a previous transfusion. Donors negative for D and c are very rare in Caucasian populations. Compatible red cell concentrates are available only from the European Bank of Frozen Blood of the Council of Europe, located at Sanquin in Amsterdam, Tthe Netherlands. The patient could be saved by requesting compatible erythrocyte concentrate from this blood bank. Severe blood loss poses a serious challenge in patients who are positive for alloantibodies against blood group antigens with a high frequency in the population, and in patients who are themselves negative for such antigens. The presence of alloantibodies is often the result of previous blood transfusions. In view of the large number of antigens on erythrocytes, one should therefore be conservative as to blood transfusion in order to prevent alloantibody formation. PMID- 16355577 TI - [A pregnant woman with irregular erythrocyte antibodies for whom no compatible packed red blood cells were available]. AB - A 45-year-old woman underwent a Caesarean section at a gestational age of over 32 weeks. Screening for irregular erythrocyte antibodies in the transfusion laboratory yielded a positive result. It appeared that the patient had for several years been known to have antibodies against At(a), a high-frequency antigen that may cause severe transfusion reactions when incompatible packed cells are administered. No autologous donated blood was available and the only compatible At(a)-negative unit of packed cells in the Blood Bank of the Council of Europe was damaged during the thawing process. A cell saver was therefore used during the Caesarean section, and family members were summoned for donation. This case report illustrates the necessity of a transfusion plan for pregnant women with (rare) irregular antibodies. PMID- 16355578 TI - [Thyrotoxicosis following the use of ashwagandha]. AB - A 32-year-old healthy woman developed thyrotoxicosis while taking capsules that contained ashwagandha herbal extract for symptoms of chronic fatigue. She was not taking any other remedies or medications. During the first few weeks, she took the capsules only occasionally without any symptoms, but after increasing the dose, she experienced clinical symptoms indicative of thyrotoxicosis. This was confirmed by laboratory assessment. The symptoms resolved spontaneously after discontinuation of the ashwagandha capsules and laboratory values normalised. To our knowledge, this relationship has not been reported previously in humans. Data from animal studies, however, have suggested that ashwagandha can increase serum concentrations of thyroid hormones. This case study suggests that thyrotoxicosis is a potentially serious side effect of ashwagandha. PMID- 16355579 TI - [The 'memory lymphocyte immunostimulation assay' (MELISA) is useless for the detection of metal allergy]. PMID- 16355580 TI - [Large-scale contact investigation for tuberculosis in Zeist]. PMID- 16355581 TI - [Restrictive dermopathy: a rare, lethal genodermatosis]. PMID- 16355583 TI - Suite solution. PMID- 16355582 TI - Younger managers; older workers. PMID- 16355584 TI - Hot new options for hospital fire safety. PMID- 16355585 TI - Braving the challenge of CBRN. PMID- 16355586 TI - Full-feature protection. PMID- 16355587 TI - Early warnings: health care preparedness. AB - As nurses, we represent the backbone of the health care system. It is essential that we have a core understanding of infectious disease emergencies and begin to use the strengths that characterize nursing. These strengths include the ability to evaluate situations and use evidence on which to base our actions. Early identification of an infectious disease emergency is one example of using nursing skills to strengthen emergency preparedness. During an infectious disease emergency, nurses certainly will bear the burden of patient management. Because of this, the need for infectious disease emergency preparedness has become a national priority and a moral imperative for all nurses. One topic necessary for ED and OH nurses' preparedness has been discussed in this article, but nurses must take the initiative to learn more about disaster preparedness and incorporate these skills into everyday practice. PMID- 16355588 TI - Toxic VOCs and confined space entry. PMID- 16355589 TI - Do the prep work. PMID- 16355590 TI - With open arms. PMID- 16355591 TI - Beware the dark side of online EHS training. PMID- 16355592 TI - Headline: hazard assessment. PMID- 16355593 TI - Verifying lockout/tagout electrically safe status. PMID- 16355594 TI - Keeping electrical safety simple. PMID- 16355595 TI - The medical miracle: more questions. PMID- 16355596 TI - Improvements in protease inhibitor-based HAART. PMID- 16355597 TI - Across a divide. PMID- 16355598 TI - Impact of HIV protease inhibitor resistance in treatment-naive populations in the united states. AB - There is concern about the increasing transmission of drug-resistant HIV to uninfected persons. Harboring a resistant variant of HIV-1 can influence the antiviral effect of the initial treatment regimen. These strains may be resistant to 1 or more of the existing classes of antiretrovirals. Use of resistance testing before initiating antiretroviral therapy to identify persons with acquired resistance is becoming more common. For these persons, as well as for highly treatment-experienced patients who may be transmitting drug-resistant HIV, the use of ritonavir-boosted protease inhibitors, possibly including investigational agents that retain activity against these isolates, may be a valuable approach to treatment. PMID- 16355599 TI - Use of acid-reducing agents in protease inhibitor-based HAART and the potential for negative treatment outcomes. AB - A number of protease inhibitors (PIs) are dependent on an acidic gastric pH for optimal drug dissolution and absorption. As a result, the potential for negative drug interactions with acid-reducing agents exists and could lead to subtnerapeutic drug concentrations, viral breakthrough, and development of drug resistance. Pharmacokinetic evaluations of a number of PIs given with acid reducing agents have been performed and show varying degrees of effect. Given the possibility of decreases in PI exposures, clinicians should be aware of the potential for a negative interaction when selecting PI-based HAART for patients taking acid-reducing agents. PMID- 16355600 TI - ADHD: strategies to unravel its genetic architecture. AB - Attention deficit/hyperactivity disorder (ADHD) is a common and impairing neuropsychiatric disorder with onset at preschool age and strong persistence over time. Its validity as a psychiatric disorder has been established according to Robins and Guze criteria. Genetic factors predominate in the etiology of ADHD. This paper summarizes the current status of genetic research into ADHD, and describes eight factors that complicate research into genetically complex disorders as ADHD. These factors are that multiple genes of small main effects are involved rather than main genes, the relevant phenotype is unknown, presence of clinical heterogeneity, presence of genetic heterogeneity, gene-environment correlation, gene-environment interaction, importance of endophenotypes, and importance of developmental factors. The further unraveling of the genetic architecture of ADHD will depend to a large extent on how well these complicating factors are handled or even used. PMID- 16355601 TI - Developmental dyslexia, learning and the cerebellum. AB - Theoretical frameworks for dyslexia must explain how the well-established phonological deficits and the literacy deficits arise. Our longstanding research programme has led to a distinctive 'twin level' framework that proposes, first, that the core deficits are well described in terms of poor skill automaticity. Second, these 'cognitive level' symptoms are attributed to abnormal cerebellar function--a 'brain-level' analysis. The evidence includes data from behavioural, imaging, neuroanatomical and learning studies. The frame-work leads to an 'ontogenetic' analysis that links cerebellar deficit at birth, via problems in articulation and working memory, to the known phonological, speed and literacy difficulties. Differences in locus of cerebellar impairment, experience and/or links to other brain regions may account for subtypes of dyslexia and possibly other developmental disorders. The automaticity/ cerebellar deficit framework provides an explicit demonstration that it is possible to explain motor, speed and phonological deficits within a unified account, integrating previously opposed approaches. PMID- 16355602 TI - Disorders of speech development: diagnostic and treatment aspects. AB - Language acquisition is a complex process relying on the well-tuned interaction of a variety of factors. Its genetic base is still little explored, but perhaps plays the most important role during the early periods of this process. In addition, neurological, cognitive and emotional abilities of the child as well as verbal stimulation from the environment are crucial. Language development disorders manifest themselves as late onset, slow progression or as erroneous course of the language development. Because of the multitude of factors involved, their aetiology is frequently difficult to determine. Diagnosis of such disorders commonly requires the cooperation of several professionals, like paediatricians, otolaryngologists, psychologists, and speech pathologists. The "late bloomer" hypothesis suggests, that up to 50% of children presenting with language problems in early years, make up for them without intervention up to age four years. Nevertheless, treatment for a language problem, as soon as it appears, is generally recommended in order to minimize adverse effects on succeeding developmental steps. PMID- 16355603 TI - Disorders of motor development (clumsy child syndrome). AB - This presentation will focus on motor competence, the clumsy child, perceptual deficits in clumsy children and possible neurological dysfunction in this group of children. Motor competence not only allows children to carry out everyday practical tasks, but it is also an important determinant of their level of self esteem and of their popularity and status in their peer group. Research has shown that about 6-10% of children have motor competences well below the norm. It is unusual for motor problems to simply disappear over time. In the absence of intervention the syndrome is likely to manifest itself. In the clinical literature attempts have been made to establish causal links between surface manifestations of clumsiness and underlying perceptual deficits. In this respect the attention is primarily directed towards the concept of inter- and intra-modal matching, particularly with respect to vision and proprioception, an ability deemed to underlie many real-life motor skills. Neurobehavioural model of inter- and intra-modal matching and deficit model is presented. Findings from studies using this paradigm are discussed and it is argued that clumsiness must be seen as a neurological dysfunction (insufficiency). PMID- 16355604 TI - Tic disorders and obsessive compulsive disorder: where is the link? AB - Over the last years evidence on the overlap between tic-disorders (TD) and obsessive compulsive behavior/disorder (OCB/OCD) has increased. The main focus of research have been the phenomenological and epidemiological similarities and differences in samples of different age, primary diagnosis (TD vs. OCD) including the co-occurrence of both. Unfortunately, only a minority of studies included all three groups (TD, TD + OCD, OCD). Nevertheless, new insight concerning possible subtypes for both TD and OCD has been gained. While some authors concentrated on OCD with/without tics we will summarize the field of TD and OCB/OCD from the viewpoint of tics, since OCB plays an important role in patients with TD. Thereby we will not only sharpen the clinicans' awareness of known differences in phenomenology, epidemiology, genetics and neurobiology, aimed to improve their diagnoses and treatment but also highlight the gaps of knowledge and discuss possibilities for further research in this field. PMID- 16355605 TI - The effectiveness of interventions for children with autism. AB - Over the past 50 years very many different treatments have been promoted as bringing about significant improvements, or even cures, for children with autism. However, few interventions involve controlled studies of any kind; randomised control trials are virtually non-existent and when appropriate research methodology has been applied the results are generally far from positive. Recent research suggests that the most effective results stem from early intensive behavioural interventions. Although many questions remain concerning the optimal age at which treatment should begin, the intensity of treatment and the many other variables that may affect outcome, there is growing evidence of general strategies that can be effective in ameliorating the problems associated with autism. PMID- 16355606 TI - Schizophrenia and related disorders in children and adolescents. AB - This paper reviews the concept and recent studies on childhood and adolescent psychoses with special reference to schizophrenia. After a short historical introduction, the definition, classification, and epidemiology of child- and adolescent-onset psychoses are described, pointing out that some early-onset psychotic states seem to be related to schizophrenia (such as infantile catatonia) and others not (such as desintegrative disorder). The frequency of childhood schizophrenia is less than 1 in 10,000 children, but there is a remarkable increase in frequency between 13 and 18 years of age. Currently, schizophrenia is diagnosed according to ICD-10 and DSM-IV criteria. The differential diagnosis includes autism, desintegrative disorder, multiplex complex developmental disorder (MCDD) respectively multiple developmental impairment (MDI), affective psychoses, Asperger syndrome, drug-induced psychosis and psychotic states caused by organic disorders. With regard to etiology, there is strong evidence for the importance of genetic factors and for neurointegrative deficits preceding the onset of the disorder. Treatment is based upon a multimodal approach including antipsychotic medication (mainly by atypical neuroleptics), psychotherapeutic measures, family-oriented measures, and specific measures of rehabilitation applied in about 30% of the patients after completion of inpatient treatment. The long-term course of childhood- and adolescent-onset schizophrenia is worse than in adulthood schizophrenia, and the patients with manifestation of the disorder below the age of 14 have a very poor prognosis. PMID- 16355607 TI - [Modernized traditional medicine could not take the place of integrative medicine]. PMID- 16355608 TI - [Effect of changji' an oral liquid on activated signal alterative intensity in algesthesia domain in patients with diarrhea type irritable bowel syndrome due to gan-pi disharmony]. AB - OBJECTIVE: To observe the effect of Changji' an (CJA) oral liquid on the activated signal alterative intensity (ASAI) in intracranial algesthesia domain in patients with diarrhea type irritable bowel syndrome (IBS) due to Gan-Pi disharmony. METHODS: Twenty-four patients were randomly divided into 2 groups, 14 in the treated group and 10 in the control group, they were administrated with CJA and placebo respectively. The sensory threshold and score in the two groups recorded by rectal inflation test were compared and analyzed. The change of ASAI in intracranial algesthesia domain was analyzed by functional magnetic resonance imagine (fM-RI) during rectum being inflated with 30 ml, 60 ml, 90 ml and 120 ml of gas respectively. RESULTS: The initial sensory thresholds in the two groups were insignificantly different, but significant difference did show between the two groups in urgent defecation threshold and pain threshold after treatment (P < 0.05). Comparison in visual simulative scores between the two groups after treatment at rectal inflated for 30 ml showed no significant difference, but it showed significant difference when the inflation was over 30 ml (P < 0.05). In the treated group, the ASAI in insula cortex when rectal inflation being 90 ml or 120 ml and that in thalamus when rectal inflation being 120 ml were significantly decreased (P < 0.05). But in the control group, it changed insignificantly after treatment. CONCLUSION: The treatment of CJA on Gan-Pi disharmony caused diarrhea type IBS might be effected by regulating the ASAI in intracranial insula cortex and thalamus. PMID- 16355609 TI - [Clinical efficacy of shenzhe zhuyun mixture in treating esophageal lesion in patients with systemic sclerosis]. AB - OBJECTIVE: To investigate the clinical efficacy of Shenzhe Zhuyun mixture (SZM) in treating esophageal lesion in patients with systemic sclerosis. METHODS: Sixty four patients conforming to the inclusion criteria were randomly divided into 2 groups, the treated group and the control group treated respectively by SZM and cisapride, with the blank preparation imitating the contrast one, the treatment course to both groups was 2 months. The clinical efficacy, the changes in symptom scores of esophageal lesion, the orthostatic and clinostatic time of barium passing through esophagus, the clinostatic esophageal emptying index of barium, and the widest transverse diameter of ectatic esophageal segment, as well as the influence of treatment on patients' quality of life were observed. RESULTS: The total effective rate in the treated group was superior to that in the control group (96.9% vs 56.2%, P < 0.01). Symptom of esophageal lesion was significantly improved after treatment in both groups (P<0.01), and the improvement in the treated group was better than that in the control group (P <0.01). The time of Barium passing through esophagus, both orthostatic and clinostatic, and the esophageal emptying index of barium were improved in the treated group (P < 0.01), but the change of the widest transverse diameter of ectatic esophagus was insignificantly improved. While in the control group, excepting the clinostatic time of barium passing through was improved (P<0.01), the other indexes showed no obvious change. Patients' quality of life was significantly improved in the treated group, but improvement didn't revealed in the control group. CONCLUSION: SZM can reinforce esophageal dynamic function in patients with systemic sclerosis, it can be an effective TCM prescription in treating esophageal lesion for them. PMID- 16355610 TI - [Study on diagnostic standard for dampness syndrome in patients with chronic gastritis]. AB - OBJECTIVE: To consummate the standard of macroscopic syndrome differentiation for diagnosis of dampness syndrome in patients with chronic gastritis (CG), based on the essentials of syndrome differentiation in TCM diagnostics and combined with the multivariant mathematical statistic method. And try to find out the methods and approaches for establishing standard for TCM syndrome differentiation. METHODS: Clinical investigation on CG patients was carried out adopting clinical epidemiological method. RESULTS: The meaningful symptoms for making diagnosis of Pi-Wei damp-heat syndrome in frequency order, were red tongue with yellow and greasy fur, sticky and greasy sensation in mouth, brown urine, constipation, dry stool, flushed face and ponderous extremities. Those for Pi deficiency with damp retention syndrome were swollen tongue with teeth-print, greasy fur, sticky and greasy sensation in mouth, tastelessness, and poor appetite. CONCLUSION: By combining the integrative medical theory with multivariant statistic method, the meaningful essentials for diagnosis of dampness syndrome can be screened out. PMID- 16355612 TI - [Study on the molecular mechanism of lingual epithelial cell apotosis and its related genes in different tongue furs]. AB - OBJECTIVE: To investigate the molecular mechanism ot lingual epithelial cell (LEC) apoptosis and its related genes expression in different tongue furs. METHODS: The LEC apoptosis and its related genes expression including bax, fas, TGF-beta 3 mRNA and protein product in tongue fur was determined using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling technique (TUNEL), in situ hybridization, immunohistochemical technique and image analysis. RESULTS: LEC apoptosis could always be seen in 4 types commonly encountered tongue fur. The tendency of changing in thickness of tongue fur was opposite to that of apoptotic index. Compared with normal thin fur, bax and fas genes were over-expressed in exfoliative fur with increased apoptosis, while in thick fur, bax and TGF-beta 3 genes were low-expressed and accompanied with decreased apoptosis. The level of apoptosis promoting genes, bax, fas, TGF beta 3 gene expression in LEC showed a tendency parallel to that of LEC apoptosis. CONCLUSION: Change of apoptosis related genes expression, bax, fas and TGF-beta 3 may effect the LEC apoptosis and be the important factor for changing of the thickness of tongue fur. PMID- 16355611 TI - [Observation on efficiency of shelian capsule as adjuvant of embolismic chemotherapy on primary hepatic carcinoma]. AB - OBJECTIVE: To evaluate the clinical efficacy and side effects of Shelian capsule (SLC) as adjuvant of embolismic chemotherapy on primary hepatic carcinoma. METHODS: One hundred and twenty patients with hepatic carcinoma were conducted arterial embolismic chemotherapy with FAM program for two therapeutic cycles. The 60 patients in the treated group were taken SLC orally for auxiliary treatment, and the other 60 in the control group were given glucurolactone instead. The tumor size, Karnofsky score, clinical symptoms, alpha fetoprotein (AFP), NK cell, T cell-subgroup and adverse effect before and after treatment between the two groups were compared respectively. RESULTS: As compared with the control group, in the treated group the Karnofsky score was better ((P < 0.05) and the remission rate of clinical symptom were superior (66.7% vs 88.3 %, P < 0.05). The ratio of CD4/CD8 and NK cell activity was significantly different between the two groups (P<0.05). There was no obvious adverse effect in the treated group. CONCLUSION: SLC can effectively enhance the immunity of patients with tumor, remit clinical symptoms, improve quality of life and without any side effects. PMID- 16355613 TI - [Effect of oxymatrine on serum matrix metalloproteinase-2 and its inhibitor in patients with chronic hepatitis B and liver cirrhosis]. AB - OBJECTIVE: To observe the effect of oxymatrine on the level of serum matrix metalloproteinase-2 (MMP-2) and its inhibitor (TIMP-2) in patients with chronic hepatitis B (CHB) and post-hepatitis B liver cirrhosis (LC), as well as on liver fibrosis indexes as hyaluronic acid (HA), laminin (LN) and IV type collagen (IV C). METHODS: Changes of all the above-mentioned indexes in patients with CHB (n = 36) and LC (n = 36) before and after treatment were determined, and the relationship of MMP-2 and TIMP-2 with liver fibrosis indexes were analyzed. RESULTS: Oxymatrine could decrease the levels of MMP-2, HA, LN and IV C in patients with severe or moderate CHB and LC of Child-pugh A, B and C grade, as compared with the data before treatment (P < 0.05). Serum level of MMP-2 and TIMP 2 was well correlated with the levels of liver cirrhosis indexes. CONCLUSION: MMP 2 and TIMP-2 can be used as a reference for evaluating the degree of LC, and oxymatrine has a certain anti-LC effect. PMID- 16355614 TI - [Clinical observation on treatment of early diabetic nephropathy by milkvetch injection combined with captopril]. AB - OBJECTIVE: To investigate the therapeutic effect of milkvetch injection (MI) combined with Captopril on early diabetic nephropathy (EDN). METHODS: A total of 69 EDN patients were randomly divided into three groups, with 23 in each group. Besides the conventional hypoglycemic therapy, patients in Group A Captopril, in Group B MI plus Captopril and in Group C MI were given respectively. The therapeutic course for all was 3 months. The related indices of EDN before and after treatment were measured and compared. RESULTS: After treatment, the blood pressure significantly lowered after treatment in Group A and B (P<0.01), but unchanged in Group C; the levels of blood glucose and HbA1c significantly decreased in Group B and C (P < 0.05 or P<0.01), and significant difference was shown in comparison of Group B with Group A (P<0.05); levels of 24hrs urinary albumin excretion (UAER), blood urea nitrogen (BUN) and serum creatinine (SCr) significantly decreased in all 3 groups, and the decrement in Group B was more significant than that in the other two groups (P <0.05). CONCLUSION: MI combined with Captopril can not only decrease blood pressure, blood glucose and HbA1c, but also significantly decrease the UAER in treating EDN. PMID- 16355615 TI - [Effect of er'bao granule on integration of ingestion behavior-related information by neurons in lateral hypothalamic area of anorexia rats]. AB - OBJECTIVE: To confirm the effect of Er'bao granule (EBG) on the sensitivity to peripheral afferent signal of neurons in lateral hypothalamic area (LHA) to illustrate the central mechanism of EBG in promoting ingestion behavior. METHODS: The anorexia rat model was established by feeding special prepared forage for one week, and all the model rats were administrated with EBG by gavage for 3 weeks. The spontaneous discharge of LHA neurons was recorded using electro-physiological extracellular recording method, and its response to electrical stimulus on gastric vagus nerve and intravenous injection of glucose were observed and compared among the normal, model and treated groups. RESULTS: As compared with the normal group, among the LHA neurons responding to afferent gastric vagal impulse, the proportion of glycemia-sensitive neurons in the model group was significantly decreased (P <0.01), but insignificant difference was shown in comparison between the treated group and the normal group. CONCLUSION: EBG play a role in regulating the sensitivity of LHA neurons to peripheral afferent signal and thus to influence the multi-afferent information integration of ingestion central neurons. PMID- 16355616 TI - [Predominant effect of bushen ningxin decoction on postmenopausal osteoporosis in mice]. AB - OBJECTIVE: To investigate the preventive and therapeutic effect of Bushen Ningxin decoction (BSNX) on postmenopausal osteoporosis. METHODS: The BALB/c female mice postmenopausal osteoporosis model was established. The model mice were treated by BSNX, with 17beta-estradiol (E2) and normal saline as positive and negative control, respectively. All mice were sacrificed after 12 weeks' treatment, the serum cytokines Th1/Th2, bone mineral density (BMD) of vertebrae (L3 - 4) and left femur were determined, and morphological quantitative analysis of bone tissue of right femurs was performed and osteoprotegerin (OPG) mRNA expression in tibia was detected by semi-quantitative RT-PCR. The ratio in weight of uterus to body was calculated, and uterine slice was gotten for histological observation. RESULTS: As compared with the negative control group, the level of interferon gamma (IFN-gamma) was significantly increased (P < 0.01) and interleukin-4 (IL-4) decreased (P < 0.05) in the BSNX treated group. The BMD of mice were improved, area of bone trabecula and OPG mRNA expression were increased in the BSNX treated and E2 treated group (P < 0.01). But the uterus in the former was significantly smaller than that in the latter (P < 0.01), while it was not significantly different to that in the negative control group. CONCLUSION: BSNX can selectively prevent and cure the postmenopausal osteoporosis, it has no or slight stimulation on uterus. The mechanism may relate with its effects in regulating the deviation of Th1/Th2, enhancing the OPG expression and inhibiting the activity of osteoclasts. PMID- 16355617 TI - [Time-phase changed character of cardiac muscle cell apoptosis and proliferation induced by angiotensin II and influences of TCM herbs for supplementing qi and activating blood circulation on it]. AB - OBJECTIVE: To observe the dynamic change of apoptosis and proliferation of cardiac muscle cells (CMC) after being induced by Angiotensin II (Ang I), and the effect of TCM herbs for supplementing qi and/ or activating blood circulation on it. METHODS: The cultured CMC of SD suckling rat were treated by Ang II, and the amplitude, rhythm and frequency of cell pulsation, the protein content, area size and apoptosis of cells at various phases as well as the influence of TCM herbs afterwards were determined by image pattern analysis system, flow cytometry and biochemical assay. RESULTS: In the model group, cell pulsation showed quickened frequency from the 24th to 48th hr after Ang II treatment with the highest amplitude at the 24th hr; the cell area enlarged at the 24th hr, the enlargement became evident at the 48hr. Cell content of protein increased at the 24th hr, which reached to its peak at the 48th hr; an increasing trend of cell number was shown from the 12th to 48th hr; cell apoptosis started to appear at the 24th hr, it increased gradually from the 48th to 72th hr, and reached to the peak at the 72th hr (P < 0.05 or P < 0.01). All the Chinese herbs, both for supplementing qi and/or activating blood circulation, especially when they were used in combination, showed favourable preventive and therapeutic effect on CMC (P < 0.05 or P < 0.01), either at the early phase (24-48th hrs) mainly manifesting hypertrophy and proliferation or the late phase (48-96th hrs) mainly manifesting apoptosis. CONCLUSION: There exist characterized phasic windows of CMC after being treated by Ang II, the window of hypertrophy-proliferation phase and the window of cell apoptosis phase. When CMC were mainly in hypertrophic manner, myocardial hypertrophy may appear. Cell apoptosis may be one of the mechanisms for turning myocardial hypertrophy to heart failure, and it could be improved by Chinese herbs for supplementing qi and/or activating blood circulation. PMID- 16355618 TI - [Effects of yinxing pingchan recipe and its components on activity of mitochondrial enzyme complex in brain of mice with Parkinson's disease]. AB - OBJECTIVE: To investigate the mechanisms of Yinxing Pingchan recipe (YXPC) and its components, i.e. the components for detoxicating (A), for calming liver (B) and for dissolving blood stasis(C), in preventing and treating Parkinson's disease, and the path of its inhibition on nigrostriatal dopaminergic neuron (DAn) apoptosis in model mice of Parkinson's disease. METHODS: Male C57BL/6J mice were divided into the normal group, the model group and four Chinese medicinal groups, that is, the YXPC group, and Group A, B and C, treated with YXPC and its components A, B and C respectively. Mouse model of Parkinson's disease was established by intraperitoneal injection with 1-methl-4-phenyl-1,2,3,6 tetrahydropyridin (MPTP). All mice were sacrificed in 2 batches at the 14th and the 28th day respectively. The activity of mitochondrial enzyme complex I, II and IV (MEC I, II and IV) in the brain of mice were measured, respectively. RESULTS: As compared with the normal group, the activity of MEC I and IV in brain was significantly lower (P < 0.05 or P < 0.01), and that of MEC II had no obvious change in the model group. As compared with the model group, the activity of MEC I was significantly higher in YXPC group and Group C at the 14th day (P < 0.05), while the activity of MECII in Group A at the 14th day, Group B at the 28th day and Group C at both 14th and 28th day was significantly lower (P<0.05 or P<0.01). Activity of MEC IV in the four Chinese medicinal groups at the 14th day all significantly increased (P<0.05 or P<0.01), and retained at high level in Group B and Group C at the 28th day (P<0.05). CONCLUSION: YXPC and its components can maintain the mitochondrial function by partial inhibiting the activity of its enzyme complex, preventing DAn apoptosis to slow down the progress of Parkinson's disease. PMID- 16355619 TI - [Effect of Salviae miltiorrhizae on mRNA expression of inducible nitric oxide synthase and organ injury in rats with severe acute pancreatitis]. AB - OBJECTIVE: To investigate the relationship between mRNA expression of inducible nitric oxide synthase (iNOS) and organ injury, and the effect of Salviae Miltiorrhizae on iNOS mRNA in severe acute pancreas (SAP) rats. METHODS: Forty five Wistar rats were randomly divided into 3 groups, the model group (MG), the Salviae Miltiorrhizae group (SMG), and the control group (CG), with 15 rats in each group. Rats in MG and SMG were established to SAP model by intraductal injection with 5% sodium taurocholate in a dose of 1.0 ml/kg, while rats in CG were merely performed sham-operation. Immediately after modeling, rats in SMG were injected with Salviae Miltiorrhizae injection (SMI) 2 ml/kg every 6 h for 4 times in total, but to rats in other two groups same volume of normal saline were administered. The level of serum amylase (AMY), nitric oxide (NO) and the volume of ascites of rats were determined 24h after modeling, and intensity of iNOS mRNA expression in pancreas, lung and kidney were detected in the same time using in situ hybridization and image analysis. The pathologic change was observed as well. RESULTS: The volume of ascites and serum levels of AMY and NO in MG were significantly higher then those in SMG and CG (P < 0.05 and P <0.01). The expression of iNOS mRNA in pancreas, lung and kidney increased in MG and SMG, it was significantly higher in MG than that in SMG. And the pathological change of pancreas, lung and kidney in SMG was milder than that in MG. CONCLUSIONS: Over expression of iNOS mRNA could cause excessive synthesis of NO, and lead to injury of pancreas, lung and kidney in SAP. Salviae Miltiorrhizae in early stage of the disease can inhibit the over-expression of iNOS mRNA to ameliorate the injury of the pancreas, lung and kidney. PMID- 16355620 TI - [Effect of mailuoning injection on calcitonin gene-related peptide expression in facial nerve of rabbits with facial spasm]. AB - OBJECTIVE: To investigate the mechanism of Mailuoning injection (MLN) in protecting facial nerve from injury. METHODS: The New Zealand white rabbit model with facial spasm was established by compressing superficial temporal artery to make artificial demyelinated lesion of the main peripheral facial nerve trunk. The successful establishment was confirmed by using electrophysiological technique to determine abnormal muscle response (AMR) which is a characteristic for facial spasm. MLN was injected continuously through ear marginal vein for 2 weeks. The change of CGRP expression in facial nerve was detected by immunohistochemical technique. RESULTS: As compared with the model group, CGRP expression in facial nerve was significantly increased in the MLN group (P <0.01), and CGRP immunoreactive positive fibers were not seen in the shamoperation group. In the model group, the facial nerve fibers degenerated obviously, myelin sheath loosened and dissociated, the turgent axons with vacuole or even completely disappeared. But the facial nerve lesion was lessened in the MLN group. CONCLUSION: MLN has a significant protective effect on facial nerve demyelination in rabbits with facial spasm, which is closely related with its effect in improving CGRP expression in the facial nerve. PMID- 16355621 TI - [Clinical observation on effect of huoxue jieyu decoction in treating depression after craniotomy: a report of 45 cases]. AB - OBJECTIVE: To observe the therapeutic effect of Huoxue Jieyu decoction (HJD) on patients with depression after craniotomy. METHODS: Ninety patients were randomly divided into 2 groups according to the sequence of their consultation. The 45 patients in the treated group were treated with conventional symptomatic treatment plus oral administration of HJD, and the 38 patients in the control group were treated in the same way but with chlorimipramine (anafranil) instead of HJD. Hamilton depression (HAMD) scores before and after treatment, occurrence of adverse effects and long-term therapeutic effect of treatment were observed. RESULTS: The score of HAMD was insignificantly different in 2 groups before and after treatment (P >0.05), but the occurrence of adverse effects between them was significantly different (P < 0.01). CONCLUSION: The therapeutic effect of HJD on depression was similar to that of anafranil, but HJD has less adverse effect and more stabilized long-term effect. PMID- 16355622 TI - [Clinical observation of fengbei huayu recipe in treating diabetic nephropathy]. AB - OBJECTIVE: To study the effects of Fengbei Huayu recipe (FHR) on urinary albumin excretion rate (UAER) in patients with diabetic nephropathy (DN). METHODS: Seventy-two type 2 DN patients in III or IV stage were randomly divided into two groups, 36 in each group. All patients were treated with conventional hypoglycemic agents and hypotensor, but those in the treated group were given additionally with FHR twice a day for 8 successive weeks. The changes of UAER, D polymer, glycosylate hemoglobin (HbA1c), renal function indexes, including blood urea nitrogen (BUN) and serum creatinine (SCr), and blood lipids, including total cholesterol (TC) and triglyceride (TG) in the two groups before and after treatment were compared. RESULTS: The levels of UAER, D-polymer, HbA1c, TC and TG were significantly decreased after treatment in the treat- ed group (P < 0.05), and the improvement were superior to those in the control group (P <0.05) respectively. But the difference of renal function before and after treatment showed no significance in both groups. CONCLUSION: FHR could not only obviously decrease the level of UAER, but also decrease the levels of blood glucose and blood-lipids in patients with DN. PMID- 16355623 TI - [Clinical observation of toothpaste containing zanthoxylum nitidum extract on dental plaque and gingivitis]. AB - OBJECTIVE: To observe the clinical efficacy of drug toothpaste containing 2% Zanthoxylum Nitidum extract in reducing accumulation of dental plaque and occurrence of gingivitis in adults. METHODS: Adopting double-blind, stratified and parallel design, the status of dental plaque and gingivitis in 200 healthy adults enrolled in the trial (100 in the treated group took the drug toothpaste to clean teeth, and 100 in the control group use common toothpaste instead) were assessed at before experiment, 3 months and 6 months after experiment. RESULTS: At the end of the 3rd month and the 6th month, as compared with the control group, the L e-Silness Gingival Index (GI) in the treated group was decreased by 9.1% (P > 0.05) and 13.9 % (P< 0.5), in the same period, the Quigley-Hein(Turesky Modification) Plaque Index (PlI) decreased by 18.7% and 22.6% (P<0.05), respectively. CONCLUSION: Toothpaste containing Zanthoxylum Nitidum extract can obviously decrease the incidence of dental plaque and enhance gingival health. PMID- 16355625 TI - [Emodin and organ fibrosis]. AB - The aim of this article was to investigate the mechanisms of emodin in antagonizing against organ fibrosis, and to illustrate that emodin can be an effective Chinese herbal preparation for treatment of organ fibrosis. PMID- 16355624 TI - [Thinking and challenge induced by the hypothesis of breaking stagnant and eliminating blood stasis in Treating acute cerebral hemorrhage by rF VII a]. AB - Definite therapeutic effect has obtained by TCM in treating acute cerebral hemorrhage (ACH) according the TCM theory of "blood circulating outside the vessels is the stasis" using breaking stagnant and eliminating blood stasis (Poxue Zhuyu) method, but no material involving the natural development of stoke in superacue stage (0 - 4 hrs after onset of the disease) being presented so far. It has been proved by randomized, double-blinded multi-centeric clinical trials that recombinant activated factor VII (rF VII a) could decreased the morbidity and disability of patients suffered from ACH, suggesting that use hemostasis treatment in ACH during superacu stage should be stressed, and the drugs for Poxue Zhuyu should be used cautiously in the period of 0 - 4 hrs after onset. The hemorrhagic disorder could be eliminated by using drugs for Poxue Zhuyu and other medicines in rational combination. PMID- 16355626 TI - [Reviewing over the history of integrative medicine in China in 20th century]. AB - Integrative traditional Chinese and Western medicine (integrative medicine) is the advanced scientific thinking system in order to settle the relationship between traditional Chinese medicine and Western medicine, which is advocated based on the convergence of traditional Chinese and Western medicine and the scientifization of traditional Chinese medicine. The establishment of the guideline of integrative medicine and the unceasing acquirement of researches are one of the light spots of Chinese medical history in the 20th century. The medical pattern of integrative medicine founded gradually in uneven pathway in the past half century reveals its tremendous advantages increasingly and should give an important impact on the progress of the Chinese medicine and even that of the world medicine. PMID- 16355627 TI - [Comments on Nigel Wiseman's A Practical Dictionary of Chinese Medicine: on the use of Western medical terms in English glossary of Chinese medicine]. AB - Mr. Wiseman believes that Western medical terms chosen as equivalents of Chinese medical terms should be the words known to all speakers and not requiring any specialist knowledge or instrumentation to understand or identify, and strictly technical Western medical terms should be avoided regardless of their conceptual conformity to the Chinese terms. According to such criteria, many inappropriate Western medical terms are selected as English equivalents by the authors of the Dictionary, and on the other hand, many ready-made appropriate Western medical terms are replaced by loan English terms with the Chinese style of word formation. The experience obtained by translating Western medical terms into Chinese when Western medicine was first introduced to China should be helpful for developing English equivalents at present. However, the authors of the Dictionary adhere to their own opinions and reject others' experience. The English terms thus created do not reflect the genuine meaning of the Chinese terms, but make the English glossary in chaos. The so-called true face of traditional Chinese revealed by such terms is merely the Chinese custom of word formation and metaphoric rhetoric. In other words, traditional Chinese medicine is not regarded as a system of medicine but merely some Oriental folklore. PMID- 16355628 TI - [Progress on the updated methods and sophisticated technics used in research of Chinese traditional medicinal compound recipes]. AB - This article reviewed the updated methods and sophisticated technics used in research of traditional Chinese medicinal compound recipes, including mainly biochip technic, computer technic, metabonomics, ADME/Tox technic, fuzzy mathematics, molecular imprinting technic, biotransformation and metabolization in intestinal bacterium and so on. PMID- 16355629 TI - [Progress in treatment of post-abdominal operational gastroparesis syndrome by integrative traditional Chinese and Western medicine]. AB - Literature regarding researches on traditional Chinese medicine therapy, Western medicine therapy and integrative traditional Chinese and Western medicine therapy of post-operational gastropareis syndrome published in recent years were reviewed in this paper. It was showed that good therapeutic effect could be obtained by applying approaches as basic Western medicine therapy, new type prokinetics and mechanical stimulation of gastroscope, etc. and in combination with traditional Chinese medicine therapies, such as orally administering, enteroclysis or acupoint drug-injection, acupuncture and abdominal compressing with Chinese drug decoction. PMID- 16355630 TI - Tort reform: a good beginning. PMID- 16355631 TI - The case for a Missouri stabilization fund. PMID- 16355632 TI - What's wrong with competition in healthcare? Isn't that the American way? PMID- 16355633 TI - Saving Medicaid. PMID- 16355634 TI - Dealing with disaster--Hurricanes Katrina & Rita. PMID- 16355635 TI - Violence in some so-called 'sports'. PMID- 16355636 TI - Protecting our assets (Missouri vs. Kansas). PMID- 16355637 TI - Physician cooperation with tort attorneys in litigatic fraud. PMID- 16355638 TI - Missouri tobacco excise tax increase initiative. PMID- 16355639 TI - Who is pain management specialist? PMID- 16355640 TI - Evidence-based drug treatment of heart failure. AB - Modern management strategies have revolutionized the treatment of patients with acute and chronic heart failure (HF). Physicians should identify the risks and benefits of each drug or interventional option being considered for the treatment of HF. Inhibition of the neurohormonal axis with angiotensin converting enzyme (ACE) inhibitors and beta-blockers has significantly improved morbidity and mortality of patients with chronic HF. An aggressive approach targeted at improving hemodynamics and end-organ function is warranted during acute exacerbations or decompensated heart failure. Immediate recognition and management of potentially reversible causes and simultaneous initiation of supportive therapy to augment the cardiac output are key elements to the successful treatment of acute heart failure. PMID- 16355641 TI - The obesity epidemic: lessons from the war on smoking. AB - Obesity is no longer a personal disease; it is a new global disease and public health phenomenon. Since the 1980s, obesity in the U.S. has taken on epidemic dimensions. This paper will review the epidemiology, mortality, and cost of obesity in the U.S. and Missouri. It will examine the national experience in the war on smoking and point out the similarities with our new struggle against obesity. Finally, it concludes with recommendations. PMID- 16355642 TI - Electrocardiographic changes in obstructive sleep apnea syndrome. AB - The acute electrocardiographic changes during apneic episodes in patients with sleep apnea are well known. Long-term electro-cardiographic changes in these patients are not well studied. We conducted a retrospective case-control study to assess the electrocardiographic changes in African-American patients with established obstructive sleep apnea syndrome (OSA). A significant percentage of patients with OSA had abnormal EKGs as compared to the control group. The effect of sleep apnea on the cardiovascular system is more complex in African-Americans due to higher prevalence of co-morbid conditions. Seventy-three percent of our patients with OSA had metabolic syndrome. PMID- 16355643 TI - Advances in liver surgery. AB - Techniques for liver surgery have evolved over the past few decades, which have, in turn, allowed for safer operations and have broadened indications for liver resection for benign and malignant liver tumors. Major advances have included appreciation of limiting liver resection to conserve functioning liver tissue and avoid extensive operations that carry a risk of postoperative liver failure, particularly in patients with cirrhosis--the so-called segment-oriented approach to liver resection; multidisciplinary management for patients with malignant liver disease to tailor treatment to each patient depending on tumor burden, condition of the liver, and associated co-morbidity; and development of laparoscopic liver resection to shorten postoperative recovery and reduce postoperative pain. Importantly, it is recommended that liver surgery, whether open or laparoscopic, be done by experienced liver surgeons in centers with the familiarity and resources to address the perioperative needs of these patients. PMID- 16355644 TI - Emergency department visits and hospitalizations resulting from dog bites, Kansas City, MO, 1998-2002. AB - Dog bites are the second most costly public health problem in the United States with half of all Americans being bitten in their lifetime. Approximately 17 to 18 percent of dog bite injuries receive medical attention. Between 1998 and 2002, Kansas City, Missouri, residents made 3,467 emergency department visits for dog bite trauma and 96 individuals were hospitalized. We report the principal diagnoses for those visits and admissions. Median hospital charges for emergency department services and hospitalizations were $300 per visit and $4,698 per admission. We discuss deficiencies in the reporting of animal bites as well as recommended community efforts in reducing the burden of dog bite trauma. PMID- 16355645 TI - A case of seven nuchal loops and a review of the literature. AB - We present a case of seven nuchal loops of umbilical cord present at birth and review the literature. We believe this is the largest number of loops reported in the literature. PMID- 16355646 TI - Tooth brushing and oral health: how frequently and when should tooth brushing be performed? AB - This review shows that there is consensus in the literature that (meticulous) tooth brushing once per day is sufficient to maintain oral health and to prevent caries and periodontal diseases. Tooth brushing is also regarded as an important vehicle for application of anti-caries agents, such as fluorides. However, most patients are not able to achieve sufficient plaque removal by performing oral hygiene measures at home. Therefore, tooth brushing twice daily is recommended by most of the dentists in order to improve plaque control. This rule is followed by most of the patients taking care for their oral health and has shown to be effective in maintenance of oral health in numerous studies. Study of the literature gives no clear evidence as to the optimal time-point of tooth brushing (before or after meals). However, in order to eliminate food impaction and to shorten the duration of sucrose impact by tooth cleaning after meals seems to be recommendable. Although--with our current knowledge of potential harm due to brushing of erosively altered and softened tooth surfaces--giving advice on a more individual basis is recommended for patients suffering from erosion. PMID- 16355647 TI - Effect of regular fluoride gel application on incipient carious lesions. AB - OBJECTIVES: To evaluate the efficacy of weekly supervised tooth-brushing with a toothpaste and a 1.23% acidulated phosphate fluoride gel (APF) gel on white lesion reversal to treat incipient enamel lesions. DESIGN: A double-blind and randomized controlled trial. SAMPLE AND METHODS: Three-hundred seven to 12-year old Brazilian schoolchildren who presented with white spots on the buccal surfaces of permanent upper incisors were randomly allocated to three groups. In group I, children underwent supervised tooth-brushing and APF gel (1.23%) for one minute once a week. In group II, children were subjected to a weekly supervised tooth-brushing and a topical application of placebo, whereas group III (control) received no intervention. An expert dental examiner performed all intra-oral examinations for dental caries (DMF-s and dmf-s) and oral hygiene (Visible Plaque Index, Gingival Blood Index). RESULTS: After three months, 258 children and 460 lesions were analyzed. The results of the clinical evaluation of arrested white spots did not differ significantly (p = 0.95) between the two experimental groups (57.9% in group I and 56.8% in group II) but differed significantly between them and the control group (p = 0.022). Logistic regression analysis identified gingival blood index (OR = 1.70, CI = 1.13 -2.55), DMF-s2 (OR = 1.61, CI = 1.07 2.43) and number of white spot lesions (OR = 1.76, CI = 1.04 to 2.98) as independent 'risk' factors for white spot lesion activities. On the other hand, supervised tooth-brushing with APF (OR = 0.55, CI = 0.34-0.91) or without APF (OR = 0.58, CI = 0.35-0.94) was a protective factor. CONCLUSIONS: These results suggest that the weekly supervised tooth-brushing was able to arrest enamel white spots and that poor oral hygiene increases the probability of keeping white spots active. PMID- 16355648 TI - Primary reason for tooth extraction in a Brazilian adult population. AB - PURPOSE: To identify the primary reasons for tooth extraction in a Brazilian adult population. MATERIALS AND METHODS: Interviews and oral examinations were conducted with 466 subjects aged from 18-76 years in the city of Maceio, Brazil. Frequency distributions, means and medians were calculated and the chi-square test was used to determine the level of significance. A logistic regression model was used to evaluate the variables associated with reasons for tooth extraction. RESULTS: Of the 466 extractions, 295 (63.3%) were due to dental caries, 61 (13.1%) due to periodontal disease, 56 (12.0%) for orthodontic reasons, 32 (6.9%) at the patient's request, 15 (3.2%) for pre-prosthetic reasons, four (0.9%) due to pericoronitis, two (0.4%) due to trauma and one (0.2%) for other reasons. Tooth extraction due to caries and other causes (excluding periodontal disease) shows a significant association with family income, toothache, type of health centre (public or private), educational level (P < 0.001) and marital status (P = 0.002). The logistic regression model has shown that patients undergoing treatment at a public health centre, those suffering from toothache or with an incomplete secondary education were more likely to lose their teeth due to caries, with educational level as the strongest indicator. For tooth extraction due to periodontal disease and other causes (excluding dental caries) the variables age, family income, type of health centre, tooth types (anterior or posterior), educational level (P < 0.001) and toothache (P = 0.006) were statistically significant, and age was the only associated variable in the logistic regression model. CONCLUSION: Dental caries was the main cause of tooth extraction in the study group, following tooth extraction due to periodontal disease. PMID- 16355649 TI - Oral retention of fluoride from a mucosa adhesive paste (Orabase) supplemented with NaF--a pilot study. AB - PURPOSE: Individuals with cognitive impairments have an increased caries risk. Methods to prevent caries based on home care use of fluorides (F) presuppose a person's ability to cooperate and follow instructions. The aim of the present investigation was therefore to develop and evaluate an F-containing mucosa adhesive paste, which can be applied in home care by nursing staff. MATERIALS AND METHODS: A cross-over, double-blind study design was used. The F concentrations were studied in whole saliva and interproximal fluid samples after application of a paste with 0.1% F based on the mucosa adhesive Orabase. A placebo product without F was used as control. In six healthy adults the paste was applied before sleep in the vestibulum in the region of the first molar in each quadrant. The application was repeated during four nights. RESULTS: Six hours after the application of the F paste the F concentrations in the interproximal areas and whole saliva had increased by 12-160 times compared to samples collected before the applications. The lower increase of the interproximal F concentrations was observed in the front region. CONCLUSIONS: The results suggest that an F containing mucosa adhesive paste may be a promising vehicle for caries prevention in subjects unable to follow traditional home care recommendations. PMID- 16355650 TI - Dental caries and ear infections in preschool-aged children. AB - PURPOSE: The purpose of this case-control study was to assess the association between ear infection and dental caries. MATERIALS AND METHODS: The sample consisted of 126 children (range: two to five years) with no major medical problems or craniofacial anomalies. Ear infection history, demographic, dental, health and diet history of each child was determined using a questionnaire administered to the parent/guardian of the child. Dental charts were used to abstract dmft (decayed, missing and filled teeth) scores for children with dental caries (DC: dmft > or = 1) and without caries (NDC: dmft = 0), and oral hygiene index (OHI) scores. RESULTS: Chi-square analysis indicated no differences in ear infection history between the 71 DC and 55 NDC (past year: 35% vs. 40%; lifetime: 30% vs. 31%) children. However, there was a trend (p = 0.07) for the mean number of ear infections to be higher in DC versus the NDC group. Baby bottle use was highest among those who had both ear infection and caries. Multivariate regression model revealed that OHI scores, reason for dental visit, and frequency of visits were the best predictors of dmft scores. CONCLUSION: An association between dental caries and ear infection was not observed in this sample. Future investigations should explore common risk factors in increasing the risk of both diseases simultaneously. PMID- 16355651 TI - Smoking and vertical bone defects in a Saudi Arabian population. AB - PURPOSE: To investigate the relationship between water pipe and cigarette smoking and the prevalence and severity of vertical periodontal bone defects. MATERIAL AND METHODS: A study sample of 355 individuals in the age range 17 to 60 years was recruited from Jeddah, Saudi Arabia. Full sets of intra-oral radiographs for each individual were assessed with regard to the presence or absence of vertical bone defects. A vertical defect was defined as an angular resorption of the interdental marginal bone of 2 mm or more at either the mesial or distal aspect of the root. RESULTS: The overall prevalence of vertical defects was 39%, with a specific prevalence of 47% in water pipe smokers, 54% in cigarette smokers, and 23% in non-smokers. The prevalence was significantly elevated in both types of smokers compared with non-smokers (p < 0.001). Expressed as the proportion of sites with vertical defects per person, the severity was 2.6% for water pipe smokers, 2.8% for cigarette smokers, and 1.3% for non-smokers. The association between smoking and severity of vertical defects was statistically significant (p < 0.001). The severity of vertical defects was significantly greater in heavy exposure compared to light exposure smokers in water pipe as well as cigarette smokers (p < 0.001). The relative risk associated with water pipe and cigarette smoking was 2.9-fold and 6.6-fold increased, respectively, compared to non smoking. CONCLUSION: The present observations suggest that prevalence and severity of vertical periodontal bone defects are increased in tobacco smokers. The association of vertical bone loss with water pipe smoking is comparable to the association with cigarette smoking. PMID- 16355652 TI - Barriers to the provision of oral health education among Mongolian dentists. AB - PURPOSE: To study barriers to providing oral health education (OHE) to their patients among Mongolian dentists. SUBJECTS AND METHODS: A questionnaire survey was carried out in 2000 among practising dentists (n = 250) in the capital city of Mongolia. The response rate was 98%. Barriers to OHE were measured by six statements describing problems that might interfere with dentists' OHE activities and attitudes towards it by a statement about its importance; all answers were rated on a 5-point Likert scale (strongly agree-strongly disagree). Self perceived competency in providing OHE was measured by means of a 4-point scale of competency (very to not at all competent) and preventive knowledge by 14 statements related to preventive dentistry. Dentists' work-related backgrounds were work experience in years, type of practice (state or private) and field of practice (general practice or speciality field). Statistical evaluation was carried out by chi-square test and logistic regression analysis. RESULTS: Poor appreciation of OHE by patients (85%) followed by insufficient teaching materials (73%) and time constraint due to the huge demand for curative care (59%) were the most commonly agreed-upon barriers. Concerning their attitudes towards and competency in providing OHE, 88% of the dentists agreed that OHE should be statutory, and 85% perceived themselves at least quite competent. Dentists' years of work experience were positively, but preventive knowledge score negatively associated with their agreeing about barriers to OHE activities. CONCLUSION: Despite appreciation of OHE, Mongolian dentists seem to face practical barriers to providing OHE activities. PMID- 16355653 TI - Etiology of xerostomia and dental caries among methamphetamine abusers. AB - This study reviews the peripheral effects of methamphetamine on the salivary acini, the pathogenesis of methamphetamine-induced xerostomia, and its anecdotal relationship to dental caries. Methamphetamine is a sympathomimetic central stimulant which is abused for its euphoric effects. Its pharmacological action is exerted indirectly by sustaining high levels of catecholamines in the synaptic cleft and directly by binding to the postsynaptic adrenergic receptors. Methamphetamine abusers report subjective perception of xerostomia, which cannot be explained by the direct peripheral action of methamphetamine on the secretory acini. The drug may cause a decrease in salivary flow rate by centrally inhibiting salivatory nuclei via stimulation of alpha-2 receptors in the brain. Drug mediated dehydration state may influence the perception of dry mouth in abusers. The decreased salivary flow rate, either due to a central inhibitory action of methamphetamine or generalised dehydration, likely contributes to the increased occurrence of dental caries. Five cases of methamphetamine abuse are presented, three of whom experienced rampant dental caries. A direct association between methamphetamine abuse and the occurrence of rampant caries was not clear. PMID- 16355654 TI - Autocalibration of a projector-camera system. AB - This paper presents a method for calibrating a projector-camera system that consists of multiple projectors (or multiple poses of a single projector), a camera, and a planar screen. We consider the problem of estimating the homography between the screen and the image plane of the camera or the screen-camera homography, in the case where there is no prior knowledge regarding the screen surface that enables the direct computation of the homography. It is assumed that the pose of each projector is unknown while its internal geometry is known. Subsequently, it is shown that the screen-camera homography can be determined from only the images projected by the projectors and then obtained by the camera, up to a transformation with four degrees of freedom. This transformation corresponds to arbitrariness in choosing a two-dimensional coordinate system on the screen surface and when this coordinate system is chosen in some manner, the screen-camera homography as well as the unknown poses of the projectors can be uniquely determined. A noniterative algorithm is presented, which computes the homography from three or more images. Several experimental results on synthetic as well as real images are shown to demonstrate the effectiveness of the method. PMID- 16355655 TI - A modified K-means algorithm for circular invariant clustering. AB - Several important pattern recognition applications are based on feature vector extraction and vector clustering. Directional patterns are commonly represented by rotation-variant vectors Fd formed from features uniformly extracted in M directions. It is often desirable that pattern recognition algorithms are invariant under pattern rotation. This paper introduces a distance measure and a K-means-based algorithm, namely, Circular K-means (CK-means) to cluster vectors containing directional information, such as Fd, in a circular-shift invariant manner. A circular shift of Fd corresponds to pattern rotation, thus, the algorithm is rotation invariant. An efficient Fourier domain representation of the proposed measure is presented to reduce computational complexity. A split and merge approach (SMCK-means), suited to the proposed CK-means technique, is proposed to reduce the possibility of converging at local minima and to estimate the correct number of clusters. Experiments performed for textural images illustrate the superior performance of the proposed algorithm for clustering directional vectors Fd, compared to the alternative approach that uses the original K-means and rotation-invariant feature vectors transformed from Fd. PMID- 16355656 TI - Clustering ensembles: models of consensus and weak partitions. AB - Clustering ensembles have emerged as a powerful method for improving both the robustness as well as the stability of unsupervised classification solutions. However, finding a consensus clustering from multiple partitions is a difficult problem that can be approached from graph-based, combinatorial, or statistical perspectives. This study extends previous research on clustering ensembles in several respects. First, we introduce a unified representation for multiple clusterings and formulate the corresponding categorical clustering problem. Second, we propose a probabilistic model of consensus using a finite mixture of multinomial distributions in a space of clusterings. A combined partition is found as a solution to the corresponding maximum-likelihood problem using the EM algorithm. Third, we define a new consensus function that is related to the classical intraclass variance criterion using the generalized mutual information definition. Finally, we demonstrate the efficacy of combining partitions generated by weak clustering algorithms that use data projections and random data splits. A simple explanatory model is offered for the behavior of combinations of such weak clustering components. Combination accuracy is analyzed as a function of several parameters that control the power and resolution of component partitions as well as the number of partitions. We also analyze clustering ensembles with incomplete information and the effect of missing cluster labels on the quality of overall consensus. Experimental results demonstrate the effectiveness of the proposed methods on several real-world data sets. PMID- 16355657 TI - Noisy text categorization. AB - This work presents categorization experiments performed over noisy texts. By noisy, we mean any text obtained through an extraction process (affected by errors) from media other than digital texts (e.g., transcriptions of speech recordings extracted with a recognition system). The performance of a categorization system over the clean and noisy (Word Error Rate between approximately 10 and approximately 50 percent) versions of the same documents is compared. The noisy texts are obtained through handwriting recognition and simulation of optical character recognition. The results show that the performance loss is acceptable for Recall values up to 60-70 percent depending on the noise sources. New measures of the extraction process performance, allowing a better explanation of the categorization results, are proposed. PMID- 16355658 TI - Matching shape sequences in video with applications in human movement analysis. AB - We present an approach for comparing two sequences of deforming shapes using both parametric models and nonparametric methods. In our approach, Kendall's definition of shape is used for feature extraction. Since the shape feature rests on a non-Euclidean manifold, we propose parametric models like the autoregressive model and autoregressive moving average model on the tangent space and demonstrate the ability of these models to capture the nature of shape deformations using experiments on gait-based human recognition. The nonparametric model is based on Dynamic Time-Warping. We suggest a modification of the Dynamic time-warping algorithm to include the nature of the non-Euclidean space in which the shape deformations take place. We also show the efficacy of this algorithm by its application to gait-based human recognition. We exploit the shape deformations of a person's silhouette as a discriminating feature and provide recognition results using the nonparametric model. Our analysis leads to some interesting observations on the role of shape and kinematics in automated gait based person authentication. PMID- 16355659 TI - Analyzing and capturing articulated hand motion in image sequences. AB - Capturing the human hand motion from video involves the estimation of the rigid global hand pose as well as the nonrigid finger articulation. The complexity induced by the high degrees of freedom of the articulated hand challenges many visual tracking techniques. For example, the particle filtering technique is plagued by the demanding requirement of a huge number of particles and the phenomenon of particle degeneracy. This paper presents a novel approach to tracking the articulated hand in video by learning and integrating natural hand motion priors. To cope with the finger articulation, this paper proposes a powerful sequential Monte Carlo tracking algorithm based on importance sampling techniques, where the importance function is based on an initial manifold model of the articulation configuration space learned from motion-captured data. In addition, this paper presents a divide-and-conquer strategy that decouples the hand poses and finger articulations and integrates them in an iterative framework to reduce the complexity of the problem. Our experiments show that this approach is effective and efficient for tracking the articulated hand. This approach can be extended to track other articulated targets. PMID- 16355660 TI - Multiple paths extraction in images using a constrained expanded trellis. AB - Single shortest path extraction algorithms have been used in a number of areas such as network flow and image analysis. In image analysis, shortest path techniques can be used for object boundary detection, crack detection, or stereo disparity estimation. Sometimes one needs to find multiple paths as opposed to a single path in a network or an image where the paths must satisfy certain constraints. In this paper, we propose a new algorithm to extract multiple paths simultaneously within an image using a constrained expanded trellis (CET) for feature extraction and object segmentation. We also give a number of application examples for our multiple paths extraction algorithm. PMID- 16355661 TI - Generalized principal component analysis (GPCA). AB - This paper presents an algebro-geometric solution to the problem of segmenting an unknown number of subspaces of unknown and varying dimensions from sample data points. We represent the subspaces with a set of homogeneous polynomials whose degree is the number of subspaces and whose derivatives at a data point give normal vectors to the subspace passing through the point. When the number of subspaces is known, we show that these polynomials can be estimated linearly from data; hence, subspace segmentation is reduced to classifying one point per subspace. We select these points optimally from the data set by minimizing certain distance function, thus dealing automatically with moderate noise in the data. A basis for the complement of each subspace is then recovered by applying standard PCA to the collection of derivatives (normal vectors). Extensions of GPCA that deal with data in a high-dimensional space and with an unknown number of subspaces are also presented. Our experiments on low-dimensional data show that GPCA outperforms existing algebraic algorithms based on polynomial factorization and provides a good initialization to iterative techniques such as K-subspaces and Expectation Maximization. We also present applications of GPCA to computer vision problems such as face clustering, temporal video segmentation, and 3D motion segmentation from point correspondences in multiple affine views. PMID- 16355662 TI - Integration of geometric elements, Euclidean relations, and motion curves for parametric shape and motion estimation. AB - This paper presents an approach to shape and motion estimation that integrates heterogeneous knowledge into a unique model-based framework. We describe the observed scenes in terms of structured geometric elements (points, line segments, rectangles, 3D corners) sharing explicitly Euclidean relationships (orthogonality, parallelism, colinearity, coplanarity). Camera trajectories are represented with adaptative models which account for the regularity of usual camera motions. Two different strategies of automatic model building lead us to reduced models for shape and motion estimation with a minimal number of parameters. These models increase the robustness to noise and occlusions, improve the reconstruction, and provide a high-level representation of the observed scene. The parameters are optimally computed within a sequential Bayesian estimation procedure that gives accurate and reliable results on synthetic and real video imagery. PMID- 16355663 TI - Effective representation using ICA for face recognition robust to local distortion and partial occlusion. AB - The performance of face recognition methods using subspace projection is directly related to the characteristics of their basis images, especially in the cases of local distortion or partial occlusion. In order for a subspace projection method to be robust to local distortion and partial occlusion, the basis images generated by the method should exhibit a part-based local representation. We propose an effective part-based local representation method named locally salient ICA (LS-ICA) method for face recognition that is robust to local distortion and partial occlusion. The LS-ICA method only employs locally salient information from important facial parts in order to maximize the benefit of applying the idea of "recognition by parts." It creates part-based local basis images by imposing additional localization constraint in the process of computing ICA architecture I basis images. We have contrasted the LS-ICA method with other part-based representations such as LNMF (Localized Nonnegative Matrix Factorization) and LFA (Local Feature Analysis). Experimental results show that the LS-ICA method performs better than PCA, ICA architecture I, ICA architecture II, LFA, and LNMF methods, especially in the cases of partial occlusions and local distortions. PMID- 16355664 TI - Face recognition using face-ARG matching. AB - In this paper, we propose a novel line feature-based face recognition algorithm. A face is represented by the Face-ARG model, where all the geometric quantities and the structural information are encoded in an Attributed Relational Graph (ARG) structure, then the partial ARG matching is done for matching Face-ARG's. Experimental results demonstrate that the proposed algorithm is quite robust to various facial expression changes, varying illumination conditions and occlusion, even when a single sample per person is given. PMID- 16355665 TI - Theoretical bounds of majority voting performance for a binary classification problem. AB - A number of earlier studies that have attempted a theoretical analysis of majority voting assume independence of the classifiers. We formulate the majority voting problem as an optimization problem with linear constraints. No assumptions on the independence of classifiers are made. For a binary classification problem, given the accuracies of the classifiers in the team, the theoretical upper and lower bounds for performance obtained by combining them through majority voting are shown to be solutions of the corresponding optimization problem. The objective function of the optimization problem is nonlinear in the case of an even number of classifiers when rejection is allowed, for the other cases the objective function is linear and hence the problem is a linear program (LP). Using the framework we provide some insights and investigate the relationship between two candidate classifier diversity measures and majority voting performance. PMID- 16355666 TI - An efficient method for the computation of Legendre moments. AB - Legendre moments are continuous moments, hence, when applied to discrete-space images, numerical approximation is involved and error occurs. This paper proposes a method to compute the exact values of the moments by mathematically integrating the Legendre polynomials over the corresponding intervals of the image pixels. Experimental results show that the values obtained match those calculated theoretically, and the image reconstructed from these moments have lower error than that of the conventional methods for the same order. Although the same set of exact Legendre moments can be obtained indirectly from the set of geometric moments, the computation time taken is much longer than the proposed method. PMID- 16355667 TI - Runaway pacemaker in a dog. PMID- 16355668 TI - The diagnostics and prognostic value of alkaline phosphatase activity in serum and peritoneal fluid from horses with acute colic. PMID- 16355669 TI - Doppler echocardiographic assessment of ventricular function--time to move to the right? PMID- 16355670 TI - Spinal shock--comparative aspects and clinical relevance. AB - Spinal shock is the loss of muscle tone and segmental spinal reflexes that develops caudal to a severe spinal cord injury. Although little recognized in the veterinary literature, spinal shock occurs in animals and is important in both accurate lesion localization and case management. In this review, we consider why spinal shock occurs and discuss the subsequent physiologic alterations, including the development of reflex hyperactivity. We also discuss differences in the expression of spinal shock between species and suggest why recovery of reflexes is relatively rapid in animals compared with humans. Finally, the implications for clinicians dealing with animals in the period immediately after severe spinal injury are considered. PMID- 16355671 TI - Renal biopsy: a retrospective study of methods and complications in 283 dogs and 65 cats. AB - Renal biopsy often is required to establish a definitive diagnosis in dogs and cats with renal disease. In this retrospective study, we determined the complications of renal biopsy as well as factors that may be associated with development of complications and procurement of adequate renal biopsy specimens in 283 dogs and 65 cats. Data extracted from medical records at 4 institutions were evaluated using logistic regression. Proteinuria was the most common indication for renal biopsy in dogs. Complications were reported in 13.4 and 18.5% of dogs and cats, respectively. The most common complication was severe hemorrhage; hydronephrosis and death were uncommon. Dogs that developed complications after renal biopsy were more likely to have been 4 to < 7 years of age and > 9 years, to weigh < or = 5 kg, and to have serum creatinine concentrations > 5 mg/dL. The majority of biopsies from both dogs (87.6%) and cats (86.2%) were considered to be of satisfactory quality. Biopsies from dogs were more likely to be of high quality if they were obtained when the patient was under general anesthesia and more likely to contain only renal cortex if they were obtained by surgery. We concluded that renal biopsy is a relatively safe procedure, with a low frequency of severe complications. Hospital practices and patient variables have the potential to impact both the quality of the specimen obtained and the rate of complications. PMID- 16355672 TI - Utility of diagnostic tests used in diagnosis of infection in dogs experimentally inoculated with a North American isolate of Leishmania infantum infantum. AB - Eight female beagles were infected with 1 x 10(7) (low dose, LD) or 2 x 10(8) (high dose, HD) promastigotes of a North American isolate of Leishmania infantum infantum (LIVT-1 strain) isolated from naturally infected Virginia Foxhounds. Two female beagles served as negative controls and 2 male beagles chronically infected (> 3 years) with Leishmania infantum chagasi were positive controls. Bone marrow (BM) and lymph node (LN) aspirates were collected every 6-8 weeks for cytologic evaluation, parasite culture, and polymerase chain reaction (PCR). Serum samples were collected monthly for determination of serologic responses by indirect fluorescent antibody test (IFAT) and diagnostic rK39 antigen. Cultures of BM and LN aspirates and cytology evaluation were consistently positive in positive control dogs during the course of study. Negative control dogs were negative on BM and LN cultures and on cytologic evaluation of aspirates. Amastigotes were present on cytological examination of BM aspirates in 2 experimentally infected dogs. Cultures of LN aspirates were positive on 22 samples, whereas BM cultures were positive on 12 samples for all dogs. IFA titers ranged from 0 to 1 :400 in experimentally infected dogs during the course of the study. Recombinant K39 immunoassay tests were consistently positive in positive control dogs and in the HD dogs by approximately 8 weeks after infection. BM PCR products were identified more consistently in the HD dogs compared with the LD dogs. Kappa statistics indicated PCR correlated better with cultures and cytology than did IFAT or the rK39 immunoassay results in the experimentally infected dogs. PMID- 16355673 TI - A comparison of the survival times of dogs treated with mitotane or trilostane for pituitary-dependent hyperadrenocorticism. AB - The survival times of 148 dogs treated for pituitary-dependent hyperadrenocorticism were studied using clinical records from 3 UK veterinary centers between 1998 and 2003. Of these animals, 123 (83.1%) were treated with trilostane, while 25 (16.9%) were treated with mitotane. Treatment groups were compared using t-tests and analysis of variance (or their nonparametric equivalents) and chi-square tests. Survival data were analyzed using Kaplan-Meier survival plots and Cox proportional hazard methods. There was no significant difference between the population attributes from each center or between treatment groups. The median survival time for animals treated with trilostane was 662 days (range 8-1,971) and for mitotane it was 708 days (range 33-1,399). There were no significant differences between the survival times for animals treated with trilostane and those treated with mitotane. In the multivariable model (including drug, center, breed group, weight, diagnostic group, and age at diagnosis), only age at diagnosis and weight were significantly negatively associated with survival. Importantly, there was no significant effect of drug choice on survival. PMID- 16355675 TI - Effect of diet and tylosin on chronic diarrhea in beagles. AB - Seven beagles in a colony of dogs had chronic diarrhea for at least 30 days. The dogs were subsequently treated with tylosin 20 mg/kg BW q24h PO for 10 days. During the treatment period, the feces became firmer but remained loose. When the treatment was discontinued, the diarrhea reappeared in 3 weeks. The feces remained abnormally loose in all dogs treated with metronidazole, trimethoprim sulfadiazine, or doxycycline and prednisone. The diet was then changed for 10 days from a highly digestible moist pet food to a dry food developed for normal adult dogs. The feces again became firmer, although still loose in some dogs. The period was then extended to 3 month, but the fecal consistency continued to fluctuate from ideal to diarrhea. The dogs were treated a 2nd time with tylosin 20 mg/kg BW q24h PO for 10 days. The feces then became significantly firmer and remained so throughout a 3-month follow-up. We conclude that the combination of diet and tylosin was more effective than either agent alone in control of chronic diarrhea. PMID- 16355674 TI - Acid-base and electrolyte abnormalities in dogs with gastrointestinal foreign bodies. AB - Gastrointestinal foreign bodies occur commonly in dogs. The objective of the study was to describe the acid-base and electrolyte abnormalities identified in dogs with gastrointestinal foreign bodies and determine if these abnormalities are related to the site or type of foreign body present. Medical records of 138 dogs were reviewed, and information on signalment, initial venous electrolyte and acid-base values, surgical findings, relevant historical information, imaging modalities used, cost of hospital visit, intra- or postoperative complications, and survival was obtained. The site of the foreign body was recorded in 94.9% of cases and the most common site was the stomach (50%), followed by the jejunum (27.5%). The foreign bodies were linear in 36.2% of cases. The most common electrolyte and acid-base abnormalities regardless of the site or type of foreign body were hypochloremia (51.2%), metabolic alkalosis (45.2%), hypokalemia (25%), and hyponatremia (20.5%). No significant association was found between electrolyte or acid-base abnormalities and the site of foreign body. Linear, as opposed to discrete, foreign bodies were more likely to be associated with a low serum sodium concentration (odds ratio, 0.85; 95% confidence interval, 0.75 0.95). Hyperlactatemia (> 2.4 mmol/L) was seen in 40.5% of dogs. A wide variety of electrolyte and acid-base derangements are found in dogs with gastrointestinal foreign bodies. Hypochloremia and metabolic alkalosis are common in these dogs. Hypochloremic, hypokalemic metabolic alkalosis is seen with both proximal and distal gastrointestinal foreign bodies. PMID- 16355676 TI - Tei index of myocardial performance applied to the right ventricle in normal dogs. AB - Right ventricular (RV) dysfunction is a cause of exercise intolerance, hypotension, syncope, and heart failure in dogs with cardiac and respiratory disorders. The study objective was to determine Doppler-derived reference values that reflect global RV function in healthy dogs. We measured systolic time intervals and an RV index of myocardial performance (IMP) in 45 healthy dogs between 8 months and 8 years of age. Pulsed-wave Doppler recordings of mitral, tricuspid, aortic, and pulmonic were acquired. Pre-ejection period (PEP), ejection time (ET), PEP/ET, and IMP were determined for both ventricles by separate cardiac cycles. Compared to the mean left ventricular (LV) IMP (0.410; 95% confidence intervals [CI] 0.378-0.442), mean RV IMP (0.250; 95% CI 0.222 0.278) was significantly smaller, and mean ET for the RV (187 millisecond [ms]; 95% CI 182-192) was significantly longer than the LV (173 ms; 95% CI 168-179). A clinically relevant correlation was not found among RV IMP and body weight, heart rate, RV ET, RV PEP, or RV PEP/ET. Calculation of LV IMP with 2 separate sample volumes yielded smaller values than from a single sample volume, with a difference in means of 0.040. We conclude that the RV IMP is relatively independent of body weight and heart rate within the ranges studied and is consistently lower than values derived from the LV in healthy dogs. This study provides additional reference values for RV function in dogs and may be useful for identification of RV dysfunction in dogs. PMID- 16355677 TI - Biochemical analysis of pericardial fluid and whole blood in dogs with pericardial effusion. AB - Studies evaluating pericardial fluid analysis in dogs to determine the etiology of pericardial effusions have yielded conflicting results. The purpose of this prospective study was to compare acid-base status, electrolyte concentrations, glucose, and lactate of pericardial fluid to peripheral blood from dogs with pericardial effusion and to compare these variables between dogs with neoplastic and nonneoplastic pericardial effusion. Acid-base status, electrolyte concentrations, glucose, hematocrit, urea nitrogen, and lactate concentrations were evaluated in peripheral blood samples and in pericardial effusion samples of 41 client-owned dogs with pericardial effusion. Common abnormal findings in the peripheral blood of dogs with pericardial effusion included hyperlactatemia (n = 38 [of 41]; 93%), hyponatremia (n = 25/41; 61%), hyperglycemia (n = 13/41; 32%), and hypermagnesemia (n = 13/41; 32%). Bicarbonate, sodium, ionized calcium, glucose, and hematocrit were all significantly lower in the pericardial fluid compared with peripheral blood, whereas lactate, chloride, and PCO2 were significantly higher in the pericardial fluid. When comparing the concentrations of variables in the pericardial fluid of dogs with neoplasia (n = 28) to those without neoplasia (n = 13), pH, bicarbonate, and chloride were significantly lower in dogs with neoplasia, whereas lactate, hematocrit, and urea nitrogen were significantly higher in the pericardial fluid of dogs with neoplasia. The difference between peripheral and pericardial glucose concentrations was significantly larger in dogs with neoplasia than in dogs without neoplasia. Although differences between variables in dogs with neoplastic and nonneoplastic pericardial effusion were documented, clinical relevance is likely limited by the degree of overlap between the 2 groups. PMID- 16355678 TI - Quantitative assessment of regional right ventricular myocardial velocities in awake dogs by Doppler tissue imaging: repeatability, reproducibility, effect of body weight and breed, and comparison with left ventricular myocardial velocities. AB - Right ventricular myocardial (RVM) motion is poorly documented. The objective of this study was to determine the variability of RVM velocities by tissue Doppler imaging (TDI) in healthy dogs (study 1), to analyze RVM motion in a large healthy canine population (study 2), and to compare the results with those obtained for the left ventricular free wall. Six healthy Beagle Dogs were monitored in study 1, and 64 healthy dogs of 14 different breeds were monitored in study 2. Velocities were recorded in 2 segments (basal and apical) of the right and left myocardial walls. In study 1, 36 TDI examinations were performed for 4 days, whereas a single TDI examination was performed on each dog in study 2. All velocity profiles included 1 positive systolic wave and 2 negative diastolic waves. The lowest intraday and interday coefficient of variation values of the right TDI variables were observed at the base (3.5-16.1%). The variability of the right apical velocities was much higher, with most coefficient of variation values > 15%. RVM velocities were higher in the basal than in the apical segments (P < .001) and were higher than the left velocities of the corresponding segment (P < .01). Body weight and breed had an effect on only a few right and left TDI variables. TDI provides a repeatable and reproducible method for evaluating basal RV function in the dog. These data also demonstrate the heterogeneity of the myocardial velocities between the left and the right ventricles and between the base and the apex. PMID- 16355679 TI - Irradiation of brain tumors in dogs with neurologic disease. AB - Radiation therapy is the treatment of choice for many primary canine brain tumors. The radiation dose tolerated by surrounding healthy brain tissue can be a limiting factor for radiation treatment and total dose as well as fractionation schedules, and volume effects may play a role in the outcome of patients undergoing radiation therapy. The purpose of this retrospective study was to evaluate the efficacy of radiation therapy in dogs with brain tumors that showed signs of neurologic disease. Forty-six dogs with brain tumors were included in the analysis. In 34 dogs, computer-generated treatment plans were available, and dose-volume data could be obtained. The totally prescribed radiation therapy doses ranged from 35 to 52.5 Gy (mean = 40.9 [SD +/- 2.91) applied in 2.5- to 4 Gy fractions (mean = 3.2). The median overall survival time calculated for deaths attributable to worsening of neurologic signs was 1,174 days (95% confidence interval [CI], 693-1,655 days). Assuming that all deaths were due to disease or treatment consequences, the median survival time was 699 days (95% CI, 589-809 days). No prognostic clinical factors such as the location or size of the tumor or neurologic signs at presentation were identified. With computerized treatment planning and accurate positioning, high doses of radiation (> 80% of the total dose) could be limited to mean relative brain volumes of 35.3% (+/- 12.6). These small volumes may decrease the probability of severe late effects such as infarction or necrosis. In this study, very few immediate or early delayed adverse effects and no late effects were noted, and quality of life was good to excellent. PMID- 16355680 TI - Differentiating benign and malignant causes of lymphocytosis in feline bone marrow. AB - Differentiation of benign and malignant causes of lymphocytosis in blood or bone marrow can be problematic. In the present study, reports of examinations of bone marrow from cats, submitted over an 8-year period, were reviewed to identify cats with increased numbers of small lymphocytes. Of 203 reports reviewed, 12 (5.9%) indicated increased numbers of small lymphocytes. Diagnoses for these cats included chronic lymphocytic leukemia (CLL; n = 2), pure red cell aplasia (PRCA; n = 4), immune-mediated hemolytic anemia (IMHA; n = 3), thymoma (n = 1), cholangiohepatitis (n = 1), and fever of unknown origin (n = 1). Several factors were identified that could be used to differentiate reactive lymphocytosis from CLL. Cats with CLL tended to be older, and lymphocytes were slightly larger and had cleaved or lobulated nuclei. Reactive lymphocytosis was associated with immune-mediated anemias and inflammatory diseases. In reactive lymphocytosis, the proliferating lymphocytes were organized into lymphoid aggregates in bone marrow and were predominately B cells. Alternatively, in CLL and thymoma, the proliferating lymphocytes were diffusely distributed and were predominately T cells. Therefore, differentiation of the causes of lymphocytosis should include evaluation of signalment, concurrent disease conditions, lymphocyte morphology, lymphocyte distribution in bone marrow, and immunophenotype. Cat age, presence of severe anemia, and evidence of inflammatory disease also should be considered. PMID- 16355681 TI - A phase I clinical trial evaluating imatinib mesylate (Gleevec) in tumor-bearing cats. AB - A phase I clinical trial evaluating the toxicity of orally-administered imatinib mesylate was performed in 9 tumor-bearing cats. Imatinib is a small molecule, tyrosine kinase inhibitor, which selectively blocks the function of overexpressed proteins associated with various malignancies. Cats included in the study had diagnoses of fibrosarcoma, squamous cell carcinoma, and mast cell tumor, and each cat was staged using CBC and serum biochemistry; urinalysis, thoracic radiographs, and abdominal ultrasonography were performed in some cats. Most cats were treated previously by surgery, radiation therapy, chemotherapy, or some combination of these treatments. None of the cats received any concurrent chemotherapy. Six cats were treated with imatinib mesylate at 1-2 mg/kg PO q24h. Dose escalations were made to 2, 4, and 10 mg/kg PO q24h in 5 cats. Two cats started therapy at 10 mg/kg PO q24h, and 1 cat started therapy at 15 mg/kg PO q24h; all 3 cats remained at these dosages. No signs of toxicity, as evaluated by CBC and serum biochemistry, were noted in 8 of the 9 cats, and minimal gastrointestinal toxicity was observed. Due to the low frequency of adverse effects, further evaluation of imatinib is ongoing at a dosage of 10 mg/kg PO q24h. PMID- 16355682 TI - Preliminary studies of serum acute-phase protein concentrations in hematologic and neoplastic diseases of the dog. AB - Serum concentrations of acute-phase proteins (APPs): haptoglobin (Hp), ceruloplasmin (Cp), serum amyloid A (SAA), and C-reactive protein (CRP) were determined in healthy dogs (n = 15) and dogs with different diseases grouped as acute inflammation (I, n = 12), hematologic neoplasias (HT, including leukemia and lymphoma, n = 16), nonhematologic neoplasias (NHT, including epithelial, mesenchymal, and mixed, n = 20), and autoimmune hemolytic anemia (AIHA, n = 8). SAA and CRP were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits, and Hp and Cp were measured using colorimetric methods, all previously validated for use in dogs. Increased concentrations of all APPs were observed in all groups of diseased dogs, but statistical significance only was observed with Hp (I, P < .001; HT, P < .05), Cp (I, P < .05; AIHA, P < .01), and CRP (I, P < .001; HT, P < .001; AIHA, CRP P < .05). High variability in individual APPs within each group of diseases was found with no significant differences between leukemia and lymphoma as well as among different types of neoplasia. The AIHA group had smaller increases in Hp, SAA, and CRP but higher concentrations of Cp. When follow-up of individual cases was possible, a decrease in APPs generally was found in cases with favorable outcome. The results of this study suggest that neoplasia and hematologic diseases such as AIHA should be considered as possible causes of mild increases in APPs in dogs. Measurement of APPs may be helpful to assess clinical evolution and monitor treatment of these processes. PMID- 16355683 TI - A comparison of traditional and quantitative analysis of acid-base and electrolyte imbalances in horses with gastrointestinal disorders. AB - The purpose of this study was to compare traditional and quantitative approaches in analysis of the acid-base and electrolyte imbalances in horses with acute gastrointestinal disorders. Venous blood samples were collected from 115 colic horses, and from 45 control animals. Horses with colic were grouped according to the clinical diagnosis into 4 categories: obstructive, ischemic, inflammatory, and diarrheic problems. Plasma electrolytes, total protein, albumin, pH, pCO2, tCO2, HCO3-, base excess, anion gap, measured strong ion difference (SIDm), nonvolatile weak buffers (A(tot)), and strong ion gap were determined in all samples. All colic horses revealed a mild but statistically significant decrease in iCa2+ concentration. Potassium levels were mildly but significantly decreased in horses with colic, except in those within the inflammatory group. Additionally, the diarrheic group revealed a mild but significant decrease in Na+, tCa, tMg, total protein, albumin, SIDm, and A(tot). Although pH was not severely altered in any colic group, 26% of the horses in the obstructive group, 74% in the ischemic group, 87% in the inflammatory group, and 22% in the diarrheic group had a metabolic imbalance. In contrast, when using the quantitative approach, 78% of the diarrheic horses revealed a metabolic imbalance consisting mainly of a strong ion acidosis and nonvolatile buffer ion alkalosis. In conclusion, mild acid-base and electrolyte disturbances were observed in horses with gastrointestinal disorders. However, the quantitative approach should be used in these animals, especially when strong ion imbalances and hypoproteinemia are detected, so that abnormalities in acid-base status are evident. PMID- 16355684 TI - In vitro efficacy of lufenuron against filamentous fungi and blood concentrations after PO administration in horses. AB - Lufenuron is a benzoylphenyl urea-derived insecticide that has been recently introduced as a novel treatment for fungal infections in horses. The purposes of this study were to determine (1) the in vitro efficacy of lufenuron against Aspergillus and Fusarium spp. and (2) the ability of lufenuron to reach efficacious blood concentrations after PO administration in horses. Fungal colonies isolated from diseased equine corneas were tested against lufenuron solutions up to 700 microg/mL. Twenty-one adult horses received 1 of 3 PO lufenuron treatment regimens: 5 mg/kg body weight (BW) q24h for 3 days, 20 mg/kg BW q24h for 3 days, or 60 mg/ kg BW q24h for 1 day. Blood samples were collected up to 96 hours after drug administration and analyzed by high-performance liquid chromatography. Statistical analyses of lufenuron blood concentrations were performed by analysis of variance and Fischer's Least Significant Difference test, with statistical significance set at P < .05. Lufenuron showed no effect on the in vitro growth of Aspergillus or Fusarium spp. Lufenuron was detected in the blood of all but 1 horse and showed no adverse effects. The maximum blood lufenuron concentration (83.5 +/- 58.7 microg/L) was lower than the concentrations proven to be ineffective in vitro in this study. Further therapeutic use of lufenuron as an antifungal agent in horses should be based on proven efficacy against specific strains of clinically relevant fungi with pharmacokinetic data demonstrating sufficient lufenuron concentrations in target tissues. PMID- 16355685 TI - Repeatability of 2 methods for assessment of insulin sensitivity and glucose dynamics in horses. AB - Both the euglycemic-hyperinsulinemic clamp (EHC) and minimal model analysis of the frequently sampled intravenous glucose tolerance test (FSIGT) have been applied for measurement of insulin sensitivity in horses. However, no published data are available on the reproducibility of these methods. Therefore, the objective of this study was to evaluate the variation and repeatability of measures of glucose dynamics and insulin sensitivity in horses derived from minimal model analysis of the FSIGT and from the EHC method. Six healthy horses underwent both the FSIGT and EHC on 2 occasions over a 4-week period, with a minimum of 5 days between tests. Coefficient of variation (CV) and intraclass correlation coefficient (ICC) were calculated for measures of glucose metabolism and insulin sensitivity derived from each test. In the EHC, insulin sensitivity, expressed as the amount of metabolized glucose (M) per unit of serum insulin (I) (M/I ratio), averaged 0.19 +/- 0.06 x 10(-4) mmol/kg/min x (pmol/L)(-1) with an average interday CV of 14.1 +/- 5.7% (range, 7-20%) and ICC of 0.74. Minimal model analysis of the FSIGT demonstrated mean insulin sensitivity (Si) of 0.49 +/ 0.17 x 10(-4)/min x (pmol/L)(-1) with an average interday CV of 23.7 +/- 11.2% (range, 9-35%) and ICC of 0.33. Mean CV and ICC for minimal model glucose effectiveness (Sg) and acute insulin response (AIRg) were, respectively, 26.4 +/- 11.2% (range 13-40%) and 0.10 and 11.7 +/- 6.5% (range 7-21%) and 0.98. Insulin sensitivity measured by the EHC has lower interday variation when compared with the minimal model estimate derived from the FSIGT. PMID- 16355686 TI - Uroperitoneum in 32 foals: influence of intravenous fluid therapy, infection, and sepsis. AB - Foals may present to a referral hospital with the primary diagnosis of uroperitoneum (UP), or they may develop UP while hospitalized for other reasons. Historical, physical, laboratory, and diagnostic variables of foals presenting with UP were compared to those developing UP while hospitalized. Emphasis was placed on the presence of electrolyte abnormalities, evidence of sepsis or infection, and development of anesthetic complications during surgical correction of the defect. Foals developing UP while in the hospital frequently had a history of dystocia and presented at a very young age (< 48 hours) with primary clinical signs compatible with intrauterine compromise or presumed hypoxic or ischemic insult with or without sepsis. Foals referred with suspected UP often had additional problems unrelated to the urinary system. These foals had hyponatremia and hyperkalemia on presentation, whereas foals receiving intravenous fluid therapy consisting of a balanced electrolyte solution did not develop the classical pattern of electrolyte abnormalities, yet a similar increase in serum creatinine and, frequently, decreasing urine production were noted. Infection was present in 63% of the foals, and 78% of foals revealed signs suggestive of sepsis or infection. Intrauterine compromise, presumed hypoxia or ischemia, and sepsis may predispose foals to development of UP. Anesthetic complications occurred in 16% of the foals undergoing surgical correction of the defect, although hyperkalemia was only present in half of the foals with anesthetic complications. PMID- 16355687 TI - Phosphorus homeostasis in dairy cows with abomasal displacement or abomasal volvulus. AB - Abnormal phosphorus homeostasis occurs in dairy cows with an abomasal displacement or volvulus. The goal of this study was to identify potential mechanisms for hypophosphatemia and hyperphosphatemia in cows with a left displaced abomasum (LDA), right displaced abomasum (RDA), or abomasal volvulus (AV). Accordingly, the results of preoperative clinicopathologic analyses for 1,368 dairy cows with an LDA (n = 1,189), RDA, or AV (n = 179) (data set 1) and for 44 cows with an AV (data set 2) were retrieved. Laboratory values were compared by Student's t-tests, and correlation and regression analyses were performed. Thirty-four percent of the animals from data set 1 (463/1,368) were hypophosphatemic (serum phosphorus concentration ([Pi]) < 1.4 mmol/L), and 9% (122/1,368) were hyperphosphatemic ([Pi] >2.3 mmol/L). Serum [Pi] was significantly lower (P < .05) in cows with an LDA (1.60 +/- 0.53 mmol/L; mean +/- SD) than in cows with an RDA or AV (1.85 +/- 0.68 mmol/L). For cows with an LDA, [Pi] was correlated with serum urea nitrogen concentration ([SUN]) (r = 0.34) and serum concentration of magnesium ([Mg]) (r = 0.20). For cows with an RDA or AV, linear correlations existed between [Pi] and [SUN] (r = 0.45), [Mg] (r = 0.43), and serum chloride concentration ([Cl]) (r = -0.27). Stepwise logistic regression analysis indicated that low [SUN] and the diagnosis of an LDA had the strongest associations with hypophosphatemia. In cows with hyperphosphatemia, [Pi] was most strongly associated with azotemia. In cows with an AV, the strongest correlations with [Pi] were found for [SUN] and serum creatinine. We conclude that hypophosphatemia in cows with an LDA is primarily due to decreased feed intake. In contrast, hyperphosphatemia in cattle with an RDA or AV appears to result from dehydration and decreased renal blood flow. PMID- 16355688 TI - Effect of age and abomasal puncture on peritoneal fluid, hematology, and serum biochemical analyses in young calves. AB - The goals of this study were to evaluate techniques for collection of peritoneal fluid from calves, establish reference ranges for fibrinogen in peritoneal fluid during the 1st month of life, and determine if abomasal puncture would alter peritoneal fluid or hematologic variables. Twenty-two healthy Holstein calves underwent 3 peritoneal fluid collections on day 1, day 15, and day 30 of age. Fibrinogen concentration in peritoneal fluid was 0.20 g/dL and 0.10 g/dL (P < .05) for day 1 and day 30, respectively, and 0.10 at day 15 (P > .05) for calves without abomasal puncture. Plasma fibrinogen concentration was 0.60 g/dL and 0.70 g/ dL (P < .05) for days 15 and 30, respectively, in calves without abomasal puncture. There were no significant differences (P < or = .05) in peritoneal fluid and peripheral blood total protein and fibrinogen concentrations, specific gravity, total and differential cell count, or erythrocyte counts between calves with or without abomasal puncture. We concluded that the reference ranges established for fibrinogen and total protein concentration are important for accurate evaluation of peritoneal fluid in calves for further comparison with similar-aged animals with gastrointestinal-tract or abdominal-cavity disease. Additionally, accidental abomasal puncture does not alter values of fibrinogen, total protein, and nucleated cell count in peritoneal fluid and does not cause apparent clinical abnormalities. PMID- 16355689 TI - Use of the D-xylose absorption test to measure abomasal emptying rate in healthy lactating Holstein-Friesian cows and in cows with left displaced abomasum or abomasal volvulus. AB - We determined the abomasal emptying rates of Holstein-Friesian cows at different stages of lactation, with left displaced abomasum (LDA), or immediately after surgical correction of LDA or abomasal volvulus (AV). D-xylose (0.5 g/kg body weight [BW], 50% solution) was injected into the abomasum in healthy cows (group 1, 4-7 days in milk [DIM], n = 7; group 2, 90-120 DIM, n = 7; group 3, > 300 DIM, n = 7) and in cows with LDA (n = 10; group 4). D-xylose was injected into the abomasum during right flank laparotomy in cows with LDA (n = 22; group 5) and cows with AV (n = 15; group 6). The time to maximal serum D-xylose concentration was used as an index of emptying rate. The abomasal emptying rates for cows in groups 1, 2, and 3 were similar, whereas emptying was slower in cows with LDA and in cows after surgical correction of LDA or AV. The abomasal emptying rate of cows with LDA was slowed to a greater extent immediately after surgery, when compared to the rate obtained before surgery. There was no difference in abomasal emptying rate immediately after surgical correction between cows with LDA or AV. The results indicate that the increased incidence of LDA in the first month of lactation is not associated with an intrinsic decrease in abomasal emptying rate in healthy cows. Our findings also demonstrate that surgical correction further slows the emptying rate in cows with LDA. PMID- 16355690 TI - Central nervous system hemangiosarcoma in a horse. PMID- 16355691 TI - Platelet dysfunction (Glanzmann's thrombasthenia) in horses. PMID- 16355692 TI - Congenital myasthenia gravis in Smooth-Haired Miniature Dachshund dogs. AB - Three 8-week-old Miniature Smooth Haired Dachshund littermates were diagnosed with myasthenia gravis based on clinical signs, results of electrophysiological testing, and response to the short-acting anticholinesterase drug, edrophonium chloride. Congenital myasthenia gravis was confirmed by the demonstration of decreased acetylcholine-receptor density in external intercostal muscle in the absence of demonstrable serum antiacetylcholine receptor antibody or antibodies complexed to acetylcholine receptors in muscle biopsy samples. Unlike most previously reported cases of congenital myasthenia gravis that are relentlessly progressive, clinical signs resolved spontaneously by 6 months of age. PMID- 16355693 TI - Bone metabolic effects of single-dose zoledronate in healthy dogs. AB - Zoledronate, an aminobisphosphonate with potent antiresorptive activity in bone, has the potential for treatment of malignant osteolysis and hypercalcemia in dogs. The purpose of this study was to evaluate the bone metabolic effects of a single dose of zoledronate in healthy dogs. Four skeletally mature, male, intact dogs received a 15-minute i.v. infusion of zoledronate at a dosage of 0.25 mg/kg. Urine N-telopeptide of type I collagen (NTx) excretion decreased significantly from baseline by 76, 63, 77, and 73% on days 7, 14, 21, and 28, respectively (P < .0125). Serum bone-specific alkaline phosphatase (bALP) decreased significantly from baseline by 36 and 42% on days 21 and 28, respectively (P < .0125). No changes were detected in indices of calcium homeostasis (ionized calcium, intact PTH, or urine calcium excretion). Single-dose i.v. zoledronate at 0.25 mg/kg appears to inhibit homeostatic osteolytic activity in healthy, skeletally mature dogs. Prospective studies should assess repeated-dose safety and activity in healthy and diseased animals of various age groups. PMID- 16355694 TI - Ambulance trust numbers face two-thirds cut by April. PMID- 16355695 TI - Crisp warns trusts as pound 250m deficit is exposed. PMID- 16355696 TI - NHS told to heed Katrina's warning. PMID- 16355697 TI - White paper task forces start work. PMID- 16355698 TI - Trusts don't know who to consult on what. PMID- 16355700 TI - Health hotel. Thanks for all the cash, but... PMID- 16355699 TI - Health hotel. Getting down to nuts and bolts. PMID- 16355701 TI - On changing lifestyle choices. PMID- 16355702 TI - Data briefing. Resource allocation. PMID- 16355703 TI - Clinical management. Failure is not an option. AB - The deadline for targets to start treatment for cancer within 31 days of a decision to treat, and 62 days of an urgent GP referral is at the end of the year. The health secretary is concerned over lack of progress towards the targets. Problems hampering progress include data collection issues and overly complicated waiting lists. Whittington Hospital and University of Leicester Hospitals trusts have made marked progress by redesigning the care pathway. PMID- 16355704 TI - Sir Nigel: PCTs must prove worth before 2008 autonomy. PMID- 16355705 TI - PCTs lament 'flawed' policy. PMID- 16355706 TI - Public flexes its muscles on healthcare outside hospitals. PMID- 16355707 TI - On them and U.S. PMID- 16355708 TI - Will PCTs be open for business? PMID- 16355709 TI - Barometer. Mental health Oct 2005. PMID- 16355710 TI - Patient centred care. When know means know. AB - Patients' understanding of health information varies hugely, is hard to predict and can widen inequality gaps. PCTs should consider appointing patient care advisers. PMID- 16355711 TI - Patient-centred care. Direct action. PMID- 16355712 TI - On coping. PMID- 16355713 TI - On Kaiser. PMID- 16355714 TI - "How are you managing with heating and lighting, herr professor?" On the publication of the Freud-Eitingon correspondence. PMID- 16355715 TI - Categories as symptoms: conceptions of love in the psychoanalytic relationship. AB - An invitation to participate in a conference, especially the phrase, "we hope that your [contribution] might deal with same-sex love," reveals the persistent, lingering effects of the creation of the categories homosexual and heterosexual. Analysis of the phrase suggests a fantasy condensed in the invitation: that a gay psychoanalyst loves and works differently than his heterosexual colleagues and that this difference is determined by his sexual orientation. Approaching the creation of these categories as a symptom of a process of repudiation and projection, the author seeks to begin identifying and sorting through the projected psychic contents that keep these categories' specious effects alive. PMID- 16355716 TI - We are driven. AB - Is metapsychology out-or in again? Is it a millstone tied around our necks-or is it an intriguing Freudian witch whom we can even dance with? Is the concept of drives an outmoded oddity-or an indispensable companion, inspiring our understanding of the patient's material and even opening new windows for further development? Can we proceed with the concept of structures and object relationships alone -or do we need the concept of drives in order to understand what these object relationships are all about? The author clearly opts for the second option in each of these pairs of alternatives. Musing on the sophisticated metapsychology debate that unsettled psychoanalysis in the United States for many years, she reviews some of the most frequently quoted objections to the concept of drives. Further, she offers an introduction to modern drive theory with the new duality of sexual and preservative drives, as well as a different concept of aggression, and explains how drives relate to structures-specifically, to the representations of self and object. PMID- 16355717 TI - Triadic reality and the capacity to love. AB - The author describes the concept of triadic reality, which he sees as a necessary prerequisite to healthy negotiation of the oedipal phase. Without positive representations of both parents individually-and, significantly, of both parents together in relation to the child-the child is hampered in her efforts to achieve a sense of triadic reality, resulting in difficulties in self representation and in object choice, as well as a limited capacity to love. A detailed case report is presented, highlighting relevant aspects of the author's work with a patient named Marielena. PMID- 16355718 TI - To repress: a note on an ambiguity of meaning. AB - An analysand used the word repress in a dream in an unusual way. In the dream, a record had been re-pressed: it was not the original disc but a copy. This manifest meaning of the word led associatively to more latent meanings. A kind of dialectical process ensued whereby, whenever the concept of repression came up, several meanings had to be considered to set the record straight. The classical way of thinking about repression had been augmented a little by this novel meaning that the analysand had stumbled on in his dream. Psychoanalytic process was enriched by this ongoing scrutiny of repression in theory and practice. PMID- 16355719 TI - On the psychoanalytic treatment of a psychotic breakdown. AB - The author discusses the four-session-a-week psychoanalysis of a patient in psychotic breakdown with outbursts of violence. The analyst's first appearance in the transference was as a "rattle" (the noise made by his shifting in his chair), which constituted undeniable evidence of corporality--first the analyst's and then the patient's--leading eventually to the awareness of there being two separate persons in the psychoanalytic relationship. This case highlights the analyst's need to function in a particular way, and to allow him- or herself to be used in a particular way, in working with very disturbed patients, where issues of the body-mind relationship and of separation from the other are often central to the analytic work. PMID- 16355720 TI - "On my way here, I passed a man with a scab": understanding a case of severe obsessive-compulsive disorder. AB - The author describes the psychoanalysis of "John" to illustrate her view of the psychodynamic organization of obsessive-compulsive disorder. She begins with Freud's recognition of the patient's sadism and of the patient's terror of the oedipal situation, which led to the regression to the anal-erotic level of psychic organization. The author then calls upon ideas from contemporary British object relations theory to describe the damage to the objects and to the sense of self in the individual's inner world that occurs as a further defense when overwhelming danger is experienced. This integrated conceptualization is clearly illustrated in John's clinical material. PMID- 16355721 TI - Reflections on cathexis. PMID- 16355722 TI - An integrated strategy for the optimization of microarray data interpretation. AB - The completion of a microarray experiment represents just a starting point toward understanding the biology of interest. A follow-up strategy is needed to fully elucidate the functional significance of microarray-derived measurements of differential expression. Given the fact that no single approach can fully unravel the fundamental biology that is typically quite complex, the follow-up strategy must be integrated at multiple levels encompassing bioinformatics, genomics, and proteomics. In this review, we discuss an integrative approach, which can be used to prioritize microarray-derived candidate genes, define their functions, and place them in the context of the biological system being studied. PMID- 16355724 TI - Testosterone-dependent transgene expression in the liver of the CAG-lacZ transgenic rat. AB - Many endogenous gene expressions in the liver are well known to be predominant in males, compared with those of females. In contrast, the fate of hepatic transgene expression between sexes is not fully understood. Here we studied whether sex hormones changed hepatic transgene expression in the ubiquitous CAG promoter driven lacZ transgenic (Tg) rat. Both sexes of CAG-lacZ Tg rats received gonadectomy. Liver biopsy was taken weekly to determine the change of transgene expression. Histological result of adult males showed mosaic lacZ expression but it was negative in adult females, while livers in neonatal stage showed comparable expression of lacZ. Other organs exhibited equal expression in both sexes. At 2 weeks after castration, lacZ expression in male liver was significantly decreased and became negative after 4 weeks while no significant difference was observed in the lacZ expression pattern in other organs. After ovariectomy, lacZ expression in female liver remained undetectable. Moreover, testosterone treatment to gonadectomized rats of both sexes could enhance lacZ expression in the liver. In summary, we report that CAG-lacZ Tg rats demonstrate sexual dimorphism of transgene expression specifically only in the liver. Testosterone administration mediated upregulation of liver lacZ expression. Our findings suggested that androgen, especially testosterone, plays an important role in the hepatic transgene expression. PMID- 16355725 TI - An investigation of factors associated with infant mortality in two midwest counties. AB - The retrospective research study was designed to investigate the relationship between infant mortality and the six variables of early prenatal care, immaturity, maternal age, Maternal education, and socioeconomic status in two midwestern counties. Data from birth and birth and death cohort records were analyzed. The Health Belief Model was used as the theoretical framework. Its constructs were explored for potential influence on infant mortality. A significant relationship existed between the six variables and infant mortality. Based on the results of this present study, the constructs of the Health Belief Model hold promise for the development of effective interventions to affect the behavior that will reduce infant mortality. PMID- 16355723 TI - Distinct gene expression profile of human mesenchymal stem cells in comparison to skin fibroblasts employing cDNA microarray analysis of 9600 genes. AB - Broad differentiation capacity has been described for mesenchymal stem cells (MSC) from human bone marrow. We sought to identify genes associated with the immature state and pluripotency of this cell type. To prove the pluripotent state of the MSC, differentiation into osteocytes, adipocytes, and chondrocytes was performed in vitro. In contrast, normal skin cells did not harbor these differentiation abilities. We compared the expression profile of human bone marrow MSC with cDNA from one primary human skin cell line as control, using a cDNA chip providing 9600 genes. The identity of all relevant genes was confirmed by direct sequencing. Data of gene array expression were corroborated employing quantitative PCR analysis. About 80 genes were differently expressed more than threefold in MSC compared to mature skin fibroblasts. Interestingly, primary human MSC were found to upregulate a number of genes important for embryogenesis such as distal-less homeo box 5, Eyes absent homolog 2, inhibitor of DNA binding 3, and LIM protein. In contrast, mesenchymal lineage genes were downregulated in MSC in comparison to skin cells. We also detected expression of some genes involved in neural development, indicating the broad differentiation capabilities of MSC. We conclude that human mesenchymal stem cells harbor an expression profile distinct from mature skin fibroblast, and genes associated with developmental processes and stem cell function are highly expressed in adult mesenchymal stem cells. PMID- 16355726 TI - Antibiotic resistance. PMID- 16355727 TI - Living with a CPAP machine. PMID- 16355728 TI - Practical nursing students partnering within the community. PMID- 16355729 TI - Health care & health insurance improvement. To: The Journal of Practical Nursing. PMID- 16355730 TI - Prostatopexy as a treatment for urethral sphincter mechanism incompetence in male dogs. AB - OBJECTIVES: The incidence of urinary incontinence due to urethral sphincter mechanism incompetence (USMI) in male dogs is relatively rare compared with the incidence in bitches, but the medical management of USMI in male dogs is less rewarding than in bitches. Attempts have been made to manage this condition surgically using either urethral bulking agents such as Teflon or by relocating the intrapelvic bladder neck to an intra-abdominal position by vas deferentopexy. This paper reports the response to prostatopexy in male dogs with USMI. METHODS: The response to prostatopexy was determined in nine severely incontinent male dogs with USMI that were followed up for periods ranging from 10 months to five years (mean 2.3 years). RESULTS: One dog was cured, four were improved, and no improvement in the frequency or degree of urinary incontinence occurred in the remaining four animals. No complications were seen in any of the dogs. CLINICAL SIGNIFICANCE: Prostatopexy may provide a further method of treating male dogs with USMI that do not respond to medical therapy. PMID- 16355731 TI - Evaluation of clinical signs and causes of lower urinary tract disease in European cats. AB - OBJECTIVES: To investigate the clinical signs and causes of lower urinary tract disease (LUTD) in 77 cats. METHODS: Cats diagnosed with LUTD over a two-year period were included in the study. RESULTS: The study population comprised 67 male and 10 female cats. Uroliths occurred in 17 of the 77 cats (22 per cent), urethral plugs in eight cats (10 per cent) and urinary tract infection in six cats (8 per cent). In 44 cats (57 per cent), no specific cause for the disease was found and they were classified as having idiopathic LUTD. In two of the 77 cats (3 per cent) no definitive diagnosis was established. Pain was less common in cats with uroliths and haematuria was more often seen in cats with urinary tract infection. At presentation, urethral obstruction was diagnosed in 45 of the 77 cats (58 per cent). CLINICAL SIGNIFICANCE: The causes of LUTD found in cats in this study are similar to those that have been previously documented, and idiopathic LUTD is the most frequent diagnosis. However, the rate of urethral obstruction, particularly in cats with idiopathic LUTD, was higher than in other reports. The cause of this difference is unknown. PMID- 16355732 TI - Acquired urinary bladder diverticulum in a dog. AB - A neutered female chow chow aged six years and five months was evaluated for dysuria. An initial diagnosis of bladder distension and atony was made following physical, ultrasonographic and radiographic examinations. The problems did not resolve with medical management and exploratory surgery revealed a large bladder diverticulum. This report describes the radiographic findings and surgical repair of the acquired bladder diverticulum. The literature available on this condition is also reviewed. PMID- 16355733 TI - Perivulvar dermatitis in a cat treated by episioplasty. AB - Perivulvar dermatitis is common in obese, older female dogs that have infantile vulvae. Removing the excessive perivulvar skin folds by episioplasty has been recommended as the treatment of choice for perivulvar dermatitis in bitches. This report describes a case of perivulvar dermatitis in a cat that was treated by episioplasty. Ten days after surgery a substantial improvement was noticed, and 67 days later the skin in the perineal/perivulvar area appeared to be normal. The cat had not been given any medication. As in bitches, episioplasty in cats can be successful in reducing perivulvar dermatitis. PMID- 16355734 TI - Unusual vascular ring anomaly associated with a persistent right aortic arch in two dogs. AB - An unusual vascular ring anomaly consisting of a persistent right aortic arch and a left ligamentum arteriosum extending from the main pulmonary artery to an aberrant left subclavian artery and left aortic arch remnant complex was identified in a German shepherd dog and a great Dane. The left subclavian artery and left aortic arch remnant complex originated at the junction between the right distal aortic arch and the descending aorta and coursed dorsal to the oesophagus in a cranial direction. The attachment of the ligamentum arteriosum to the aberrant left subclavian artery was approximately 5 cm cranial to the point of origin of the aberrant left subclavian artery and left aortic arch remnant complex from the descending aorta in both dogs. This anomaly observed in both dogs is similar to an anomaly reported in humans, in which a persistent right aortic arch is found in conjunction with an aberrant left subclavian artery and a left aortic arch remnant (Kommerell's diverticulum). Surgical ligation and division of the left ligamentum arteriosum in both dogs, along with division of the left subclavian artery in the great Dane, resulted in resolution of clinical signs in both of the dogs in this report. PMID- 16355735 TI - Phaeochromocytoma treated by en bloc resection including the suprarenal caudal vena cava in a dog. AB - A 10-year-old, neutered male Cairn terrier was examined after experiencing intermittent vomiting of several months' duration and sudden onset of acute depression. Abdominal ultrasound examination revealed an adrenal mass invading the suprarenal part of the caudal vena cava. The vascular invasion could not be corrected so en bloc ligation/excision of the adrenal neoplasm and the suprarenal part of the vena cava was performed. Microscopic examination identified an adrenal phaeochromocytoma. The dog recovered well and had no significant vascular or renal dysfunction at follow-up 20 months after surgery. To the authors' knowledge, this is the first report of a dog surviving long term after resection of the suprarenal caudal vena cava. PMID- 16355736 TI - Radiographic features of two cases of feline proximal femoral dysplasia. AB - Congenital or acquired feline femoral dysplasia have seldom been reported in the literature. This report describes two cases of femoral dysplasia involving the proximal epiphyses. Bilateral agenesis of the femoral head and neck (case 1) constituted an incidental finding during a post-traumatic radiographic examination, while right proximal femoral dysplasia (case 2) was observed during an orthopaedic examination for lameness. To the authors' knowledge, such imaging findings have not been reported before in cats. PMID- 16355737 TI - What is your diagnosis? Synovial sarcoma. PMID- 16355738 TI - Advances in small animal fracture repair. PMID- 16355739 TI - Interactive art and science. PMID- 16355740 TI - The perception of distances and spatial relationships in natural outdoor environments. AB - The ability of observers to perceive distances and spatial relationships in outdoor environments was investigated in two experiments. In experiment 1, the observers adjusted triangular configurations to appear equilateral, while in experiment 2, they adjusted the depth of triangles to match their base width. The results of both experiments revealed that there are large individual differences in how observers perceive distances in outdoor settings. The observers' judgments were greatly affected by the particular task they were asked to perform. The observers who had shown no evidence of perceptual distortions in experiment 1 (with binocular vision) demonstrated large perceptual distortions in experiment 2 when the task was changed to match distances in depth to frontal distances perpendicular to the observers' line of sight. Considered as a whole, the results indicate that there is no single relationship between physical and perceived space that is consistent with observers' judgments of distances in ordinary outdoor contexts. PMID- 16355741 TI - Spatial and temporal properties of stereoscopic surface interpolation. AB - It is well established that under a wide range of conditions when a sparse collection of texture elements varies smoothly in depth, the spaces between the elements are assigned depth values. This disparity interpolation process has been studied in an effort to define some of its fundamental spatial and temporal constraints. To assess disparity interpolation we employed two tasks: a novel task that relies on the bisection of illusory boundaries created when subjective stereoscopic surfaces intersect, and one that relies on a 3-D shape discrimination. The results of both experiments show that there is no improvement in performance when texture density is increased from near 0.20 to 0.85 or when exposure duration is increased from 50-100 to 1000 ms. This lack of dependence on the addition of features that define the interpolated surface, along with the abrupt decline in performance below a critical value, is consistent with the view that surface interpolation is an important function of human stereoscopic vision. PMID- 16355742 TI - Do rotation coordinates provide the substrate for a mental protractor? AB - In previous studies, we have found that the accuracy in judging collinearity of lines or dots varies considerably from one subject to another as a function of the relative angle of the stimulus elements. A model of errors generally shows large excursions across several subranges of angular position. These do not appear to be motor errors, at least not ones that are well separated from perceptual mechanisms. The errors are most likely generated at primary visual cortex, or beyond. We examined and modeled accuracy in judging collinearity of dot pairs, varying the angular position of the dots through 360 degrees, the distance between the dots (stimulus span), and the distance at which the subject was required to respond (response span). Subjects manifested idiosyncratic profiles of error across angular positions, as reported previously. But across the tested range of spans, from 4 to 8 deg, the errors tended to be the same, irrespective of stimulus or response span. This suggests that the judgments are based on a radial (angular) measure of spatial position. We discuss these results in the context of proposals that the brain maps spatial position using rotation coordinates. These new data are consistent with the hypothesis that subjects use the z-axis coordinates as a mental protractor for judging angular position and collinearity. PMID- 16355743 TI - The Hermann grid illusion revisited. AB - The Hermann grid illusion consists of smudges perceived at the intersections of a white grid presented on a black background. In 1960 the effect was first explained by a theory advanced by Baumgartner suggesting the illusory effect is due to differences in the discharge characteristics of retinal ganglion cells when their receptive fields fall along the intersections versus when they fall along non-intersecting regions of the grid. Since then, others have claimed that this theory might not be adequate, suggesting that a model based on cortical mechanisms is necessary [Lingelbach et al, 1985 Perception 14(1) A7; Spillmann, 1994 Perception 23 691 708; Geier et al, 2004 Perception 33 Supplement, 53; Westheimer, 2004 Vision Research 44 2457 2465]. We present in this paper the following evidence to show that the retinal ganglion cell theory is untenable: (i) varying the makeup of the grid in a manner that does not materially affect the putative differential responses of the ganglion cells can reduce or eliminate the illusory effect; (ii) varying the grid such as to affect the putative differential responses of the ganglion cells does not eliminate the illusory effect; and (iii) the actual spatial layout of the retinal ganglion cell receptive fields is other than that assumed by the theory. To account for the Hermann grid illusion we propose an alternative theory according to which the illusory effect is brought about by the manner in which S1 type simple cells (as defined by Schiller et al, 1976 Journal of Neurophysiology 39 1320-1333) in primary visual cortex respond to the grid. This theory adequately handles many of the facts delineated in this paper. PMID- 16355744 TI - Dissociation between visual perception of allocentric distance and visually directed walking of its extent. AB - Walking without vision to previously viewed targets was compared with visual perception of allocentric distance in two experiments. Experimental evidence had shown that physically equal distances in a sagittal plane on the ground were perceptually underestimated as compared with those in a frontoparallel plane, even under full-cue conditions. In spite of this perceptual anisotropy of space, Loomis et al (1992 Journal of Experimental Psychology. Human Perception and Performance 18 906-921) found that subjects could match both types of distances in a blind-walking task. In experiment 1 of the present study, subjects were required to reproduce the extent of allocentric distance between two targets by either walking towards the targets, or by walking in a direction incompatible with the locations of the targets. The latter condition required subjects to derive an accurate allocentric distance from information based on the perceived locations of the two targets. The walked distance in the two conditions was almost identical whether the two targets were presented in depth (depth presentation condition) or in the frontoparallel plane (width-presentation condition). The results of a perceptual-matching task showed that the depth distances had to be much greater than the width distances in order to be judged to be equal in length (depth compression). In experiment 2, subjects were required to reproduce the extent of allocentric distance from the viewing point by blindly walking in a direction other than toward the targets. The walked distance in the depth-presentation condition was shorter than that in the width presentation condition. This anisotropy in motor responses, however, was mainly caused by apparent overestimation of length oriented in width, not by depth compression. In addition, the walked distances were much better scaled than those in experiment 1. These results suggest that the perceptual and motor systems share a common representation of the location of targets, whereas a dissociation in allocentric distance exists between the two systems in full-cue conditions. PMID- 16355745 TI - Homage to Peter Thompson: the Tony Blair illusion. PMID- 16355746 TI - Effects of feedforward and feedback consistency on reading and spelling in dyslexia. AB - We investigated the effects of rime consistency on reading and spelling among dyslexic children and a group of matched reading age skilled readers by manipulating consistency of orthography-tophonology (OP) mappings and consistency of mappings from phonology-to-orthography (PO). For both dyslexic and control children we found feedforward consistency effects on reading (O->P) and spelling (P->O) and feedback (O->P) consistency effects on spelling. Dyslexic children demonstrated feedback (PO) consistency effects in reading but control children did not. Our results challenge accounts of reading and spelling in dyslexia that assume feedforward consistency effects alone. We consider the implications of these results in relation to theories in which children may assess candidate responses for goodness of fit to prior expectations. We discuss the wider implications of our results for the assessment and remediation of dyslexia. PMID- 16355747 TI - Exploring dyslexics' phonological deficit I: lexical vs sub-lexical and input vs output processes. AB - We report a series of experiments designed to explore the locus of the phonological deficit in dyslexia. Phonological processing of dyslexic adults is compared to that of age- and IQ-matched controls. Dyslexics' impaired performance on tasks involving nonwords suggests that sub-lexical phonological representations are deficient. Contrasting nonword repetition vs auditory nonword discrimination suggests that dyslexics are specifically impaired in input phonological processing. These data are compatible with the hypothesis that the deficit initially affects input sub-lexical processes, and further spreads to output and lexical processes in the course of language acquisition. Further longitudinal research is required to confirm this scenario as well as to tease apart the role of the quality of phonological representations from that of verbal short-term memory processes. PMID- 16355748 TI - Phonological processing skills as predictors of literacy amongst Arabic speaking Bahraini children. AB - This paper reports a study of the reading and spelling skills of grades 1-3 Arabic-speaking children in Bahrain. Children were tested on their literacy skills (single word reading and spelling), their ability to decode letter strings (non-word reading) and measures of phonological awareness, short-term memory, speed of processing and non-verbal ability. These tests were included to identify the best predictors of literacy skills amongst Arabic young readers. The results were consistent with the literature based on tests of English-speaking children in that measures of phonological skills (decoding and awareness) were the best predictors of variability in reading and spelling among the Bahraini children. The results are discussed in terms of the literacy experiences of the children and the use of short vowels in Arabic writing. PMID- 16355749 TI - Auditory and speech processing and reading development in Chinese school children: behavioural and ERP evidence. AB - By measuring behavioural performance and event-related potentials (ERPs) this study investigated the extent to which Chinese school children's reading development is influenced by their skills in auditory, speech, and temporal processing. In Experiment 1, 102 normal school children's performance in pure tone temporal order judgment, tone frequency discrimination, temporal interval discrimination and composite tone pattern discrimination was measured. Results showed that children's auditory processing skills correlated significantly with their reading fluency, phonological awareness, word naming latency, and the number of Chinese characters learned. Regression analyses found that tone temporal order judgment, temporal interval discrimination and composite tone pattern discrimination could account for 32% of variance in phonological awareness. Controlling for the effect of phonological awareness, auditory processing measures still contributed significantly to variance in reading fluency and character naming. In Experiment 2, mismatch negativities (MMN) in event-related brain potentials were recorded from dyslexic children and the matched normal children, while these children listened passively to Chinese syllables and auditory stimuli composed of pure tones. The two groups of children did not differ in MMN to stimuli deviated in pure tone frequency and Chinese lexical tones. But dyslexic children showed smaller MMN to stimuli deviated in initial consonants or vowels of Chinese syllables and to stimuli deviated in temporal information of composite tone patterns. These results suggested that Chinese dyslexic children have deficits in auditory temporal processing as well as in linguistic processing and that auditory and temporal processing is possibly as important to reading development of children in a logographic writing system as in an alphabetic system. PMID- 16355750 TI - The subtypes of developmental dyslexia in Chinese: evidence from three cases. AB - This study investigated subtypes of developmental dyslexia in Chinese by assessing three cases of Chinese dyslexic children. A battery of screening measures, a character naming and meaning task, and metalinguistic awareness tasks were administered to each child. One of the three children demonstrated characteristics of developmental surface dyslexia, and the other two showed characteristics of developmental deep dyslexia. Moreover, the children's reading problems were found to be specifically related to their deficits in metalinguistic awareness. The dissociation between developmental surface and deep dyslexia provides support to Weekes et al.'s (Neurocase 1997; 3: 51-60) model that a semantic and a nonsemantic pathway exist independently in Chinese reading. The results also suggest that deficits in morphological and phonological awareness may cause developmental delays in the semantic and the nonsemantic pathway. PMID- 16355751 TI - Meeting the pay-for-performance imperative. PMID- 16355752 TI - 'The mission is still the mission'. PMID- 16355753 TI - Smart strategic planning for cardiovascular services. AB - Strategic planning for cardiovascular services should include formation of a cardiovascular advisory committee composed of key stakeholders. The strategic plan should include an internal assessment, external market analysis, review of operations, development of strategies and initiatives, and a financial analysis. The organization's mission and vision, as well as its financial situation, need to be considered in formulating strategies. PMID- 16355754 TI - Breaking the code. AB - University Physician Associates of New Jersey learned a number of lessons in implementing electronic charge capture: Using informal physician leaders to support the training process helped encourage adoption of new technology. Training was best accomplished in a group setting, where physician leaders could serve as role models. Approaching the project incrementally helped to achieve reasonable comfort levels. PMID- 16355755 TI - Are you keeping an eye on your organization's financial pulse? PMID- 16355756 TI - No small change: payment trends call for big preparations for 2006. AB - Payment trends that hospitals will need to respond to in 2006 are related to five areas of concern: The need to audit commercial insurers' payment for accuracy. The need for contract modeling and data-driven organization. The shift of volume risk from payers to hospitals. The shift of cost risk from payers to hospitals. The rise of consumer-directed health plans and related pay-for-performance and tiered pricing initiatives. PMID- 16355757 TI - The next big thing: health care when the patient pays. AB - With consumer-directed health plans growing in popularity, healthcare organizations should be planning: How to respond to demands for point-of-service payment. How to address competition posed by convenient care centers with attractive pricing advantages. How to offer convenient, high-quality, competitively priced care for those who have a choice. PMID- 16355758 TI - Dealing with self-pay patients--compassionately. AB - Efforts to collect money should start at the beginning--in preregistration and registration. Hospitals should be firm, but compassionate, about communicating payment responsibilities to self-pay patients. Involving all revenue cycle employees in improving the collection process will gain valuable input and ensure their buy-in. PMID- 16355759 TI - Capturing the benefits of web-based connectivity. AB - For providers as well as health plans, there are several benefits to web-based connectivity: Faster transactions. Cost savings. Heightened productivity. Increased patient and provider satisfaction. PMID- 16355760 TI - Minding your business. AB - There are a number of ways that hospitals can enhance financial perfomance from their outpatient physical therapy services: Provide training for therapists on Medicare's "eight minute rule". Differentiate between service-based and time based CPT codes on the charge ticket. Have a professional review CPT documentation before charges are submitted. Implement a program to prevent and manage no-shows and cancellations. Match staffing level with productivity. Pay careful attention to detail when documenting plans of care and preparing daily treatment notes. PMID- 16355761 TI - Leadership and power. PMID- 16355762 TI - Demystifying 'digital hospitals'. AB - Going digital requires a well-formulated strategy implementation plan. This plan is not just a list of applications to be implemented. Rather, it identifies the governance, education, communication, planning, training, and other decisions that need to be made, as well as the timing and sequencing of these components. PMID- 16355764 TI - U.S. medical liability trend showing strong signs of improvement. PMID- 16355763 TI - Revenue growth: how to win the war for markets and share. PMID- 16355765 TI - [Effects of soil temperature and humidity on soil respiration rate under Pinus sylvestriformis forest]. AB - Employing root-wrenching method and LI-6400-09 soil respiration chamber, this paper measured the diurnal changes of soil respiration rate with and without roots in situ on June 17, August 5, and October 10, 2003. The seasonal changes of soil respiration were also measured from May to September, 2004. The results showed that both the total and the root-wrenched soil respiration appeared single diurnal pattern, with the peaks presented during 12:00-14:00. The diurnal fluctuation of soil respiration on August 5 was smaller than that on June 17 and October 10. There were also obvious seasonal changes in total and root-wrenched soil respiration, as well as in root respiration, which were higher from June to August but lower in May and September. The average total soil respiration, root wrenched soil respiration, and root respiration were 3.12, 1.94 and 1.18 micromol CO2 x m(-2) s(-1), respectively, and the contribution of roots to total soil respiration ranged from 26.5% to 52.6% from May to September, 2004. There were exponential correlations between respiration rate and soil temperature, and linear correlations between respiration rate and soil humidity. The Q10 values were 2.44, 2.55 and 2.27 for total soil respiration, root-wrenched soil respiration, and root respiration, respectively. The effect of soil temperature on root-wrenched soil respiration was lager than that on total soil respiration and root respiration. Soil humidity had a larger effect on total soil respiration than on root respiration and root-wrenched soil respiration. PMID- 16355766 TI - [Dynamic changes of soil ecological factors in Ziwuling secondary forest area under human disturbance]. AB - As a widespread natural phenomenon, disturbance is considered as a discrete event occurred in natural ecosystems at various spatial and temporal scales. The occurrence of disturbance directly affects the structure, function and dynamics of ecosystems. Forest logging and forestland assart, the common human disturbances in forest area, have caused the dynamic changes of forest soil ecological factors in a relatively consistent environment. A study on the dynamics of soil bulk density, soil organic matter, soil microbes and other soil ecological factors under different human disturbance (logging and assart, logging but without assart, control) were conducted in the Ziwuling secondary forest area. The results indicated that human disturbance had a deep impact on the soil ecological factors, with soil physical and chemical properties become bad, soil organic matter decreased from 2.2% to 0.8%, and soil stable aggregates dropped more than 30%. The quantity of soil microbes decreased sharply with enhanced human disturbance. Soil organic matter and soil microbes decreased more than 50% and 90%, respectively, and soil bulk density increased from 0.9 to 1.21 g x cm( 3) with increasing soil depth. Ditch edge level also affected the dynamics of soil factors under the same disturbance, with a better soil ecological condition at low-than at high ditch edge level. PMID- 16355767 TI - [Spatial heterogeneity of soil moisture after raining at forest-grassland landscape boundary in hilly area of Loess Plateau]. AB - Soil moisture is a main factor limiting vegetation restoration in semi-arid region. In this paper, the spatial variability of different layers soil moisture after raining at the forest-grassland boundary in hilly area of Loess Plateau were studied by traditional and geostatistical analysis methods. The results showed that the moisture content in surface (0 - 10 cm) and subsurface soil layer (10 - 20 cm) of grassland was higher than that of forestland. The two layers soil moisture content at forest-grassland boundary showed a small variation but an obvious ecological distribution. By using moving split-window techniques, it was obtained that the width of edge influence in surface and subsurface layer was 8 and 6 m, respectively. Geostatistical analyses showed that the spatial distribution of two layers soil moisture had a pure nugget effect in grassland, linear model in forestland, and spherical model in forest-grassland boundary. The spatial heterogeneity of two layers soil moisture was higher at forest-grassland boundary than at forestland and grassland, which had a stronger spatial dependence and autorelation. Kriging maps expressed the spatial structural characters. The distribution of soil moisture in two layers showed a strip shape near forest edge, and a patch shape far from the edge. PMID- 16355768 TI - [Generation mechanism of woodland runoff and sediment on Loess Plateau under hypo rainfall--a case study of artificial P. tabulaeformis and secondary natural P. dadidiana stands]. AB - Based on the long-term observation and from the viewpoints of water balance and runoff-and sediment generation, this paper studied the generation processes of runoff and sediment on two typical woodlands, artificial P. tabulaeformis and secondary natural P. dadidiana, and uncultivated slope-land in Loess Plateau under hypo-rainfall. The results showed that within the range of 5.0 - 50.0 mm rainfall, the total interception of canopy and litter was 15.45 % - 56.80 % for P. tabulaeformis and 20.56% - 47.81% for P. dadidiana, and decreased with increasing rainfall. Woodlands had a higher soil water infiltration capacity than uncultivated slope-land, especially in 0-20 cm soil layer. Both the two woodlands did not generate runoff under regular rainfall. Under the assumed rainfall of 2.5 mm x min(-1) intensity and 30 min duration, P. dadidiana stand did not produce runoff, but the runoff velocity and sediment-carrying capacity of uncultivated slope-land were 23.5 times, and runoff shearing stress and energy were 8 times as much as P. tabulaeformis stand. The runoff-and sediment generation on P. tabulaeformis stand decreased by 87.6% and 99.4%, respectively, compared with those on uncultivated slopeland, which was well accorded with the average observed value in runoff plots during 1988 - 2000. The theoretical analysis on the generation mechanism of woodland runoff and sediment may be effective to evaluate the benefits of forest in soil and water conservation. PMID- 16355769 TI - [Response of sediment production to land-use change in Luergou watershed of Loess Plateau]. AB - The study on the effects of different land use patterns on the sediment production in Luergou watershed of Loess Plateau indicated that in comparing with 1982, the areas of forest and grass and of terrace in 1989 increased by 5% and 2.09%, respectively, while that of slope plowland decreased by 1.5%, with the area of naked land and shrubbery respectively increased and decreased a little. During 1998 - 2000, with the increase of vegetation cover and the decrease of steep slope plowland, the sediment production reduced significantly under high precipitation, 85326 and 52937 t under the precipitation of 803 and 786 mm, respectively, and not distinguishable in dry years. The reduction effect had a strong seasonality, which consisted with rainfall distribution. For example, in a month with 50 mm precipitation, the mean daily sediment concentration was reduced by 6 kg x m(-3), while in a month with 100 mm precipitation, the reduction was 12 kg x m(-3). Rainfall intensity also played an important role in soil erosion and sediment production, regardless of land use conditions. PMID- 16355770 TI - [Effects of forest vegetation on runoff and sediment production on sloping lands of Loess area]. AB - Based on the 1985 - 2003 fixed-position data in 9 runoff plots of Caijiachuan watershed in the Jixian county of Shanxi Province in loess area, this paper discussed the relationships between vegetation and runoff and sediment production on sloping lands of loess area. The results showed that natural-secondary forest had better function in soil and water conservation than artifical Robinia pseudoacacia forest, with runoff and sediment produced 65% - 82% and 23% - 92%, respectively. Multiple regression analysis indicated that runoff and sediment production had a significant correlation with rainfall and its intensity, but this relationship was decreased gradually with increasing canopy density. Different land-use type had different runoff and sediment production, e. g., Ostryopsis davidiana and natural-secondary forests had the least runoff and sediment production, artificial Robinia pseudoacacia and Pinus tabulaeformis forests had 5 folds of it as much as Ostryopsis davidiana forest, mixed apple trees and crops had 17.14 and 3.96 folds of it than Ostryopsis davidiana forest, respectively, while high-standard soil preparation could decrease the production obviously. Gray correlation analysis suggested that the stand canopy density and the biomass of herb and litter were the most important factors affecting the runoff and sediment production on sloping land, whose gray correlation degrees all exceeded 0.6. Mixed forest with multi-layer stand structure and shrub forest should be developed in vegetation re-construction of loess area, which could help to increase the coverage and litter thickness to dramatically decrease the runoff and sediment production on sloping land. PMID- 16355771 TI - [Mechanism of watershed soil erosion control by vegetation]. AB - From the view of hydrodynamics, this paper studied the acting mechanism of tree, grass and forest litter on slope runoff velocity and kinetic energy. The results showed that slope runoff head loss was related to slope gradient, forest density, net rainfall intensity and slope length. The relationship of water head loss with the distance among trees and the diameter at the ground of tree was Eoc (D/b)4/3. The grass on slope turned to be curved with s flowing, and thus, increased the bottom resistance of flow, and reduced the shearing stress of soil surface. Therefore, silt-carrying capacity decreased dramatically. The analysis of actually measured materials of each rainfall, runoff and sediment, and the comparison of Qiaozi eastern gully and Qiaozi west gully in Tianshui city of Gansu Province showed that under same precipitation condition, the runoff, sediment yield, flood peak discharge and maximum sediment transport rate in treated watershed was less than those in untreated watershed, suggesting that vegetation was obviously beneficial to water reservation and water and soil conservation. PMID- 16355772 TI - [Soil properties and water holding capacites of Michelia macclurei, Schima superba and Castanopis fissa stands]. AB - The study showed that the soil density, total porosity, natural water capacity and capillary moisture capacity were 1.19 g x cm(-3), 56.73%, 15.7% and 43.2% in Michelia nacclurei stand, 1.26 g x cm(-3), 54.18%, 13.0% and 37.8% in Schima superb stand, and 1.06 g x cm(-3), 60.74%, 19.4%, and 45.8% in Castanopsis fissa stand, respectively. Soil water holding capacity and aeration were good in Castanopsis fissa stand but bad in Schima superba stand, whereas Michelia macclurei stand had a medium water holding capacity and a bad aeration. The water holding capacity of litter per tree was in order of Michelia nuacclurei (20 kg) > Castanopsis fissa (15 kg) > Schima superba (8 kg), whereas that of litter in stand was Castanopsis fissa (17 t x hm(-2)) > Michelia macclurei (16 t x hm(-2)) > Schima superba (13 t x hm(-2)). The nutrient storage of litter per tree was 112.71, 31.20 and 87.30 g in Michelia macclurei, Schima superba and Castanopsis fissa stands, respectively, and that of litter in stand was 84.35,51.86 and 98.11 kg x hm(-2), respectively. The soil in the three stands was strong acidic, and the content of soil organic matter, total N, total P, total K, alkalized N, available P and available K was 18.43 g x kg(-1), 0.69 g x kg(-1), 0.17 g x kg( 1), 5.83 g x kg(-1), 45.67 mg x kg(-1), 0.83 mg x kg(-1) and 23.13 mg x kg(-1) in Michelia macclurei stand, 13.40 g x kg(-1), 0.68 g x kg(-1), 0.20 g x kg(-1), 12.32 g x kg(-1), 40.78 mg x kg(-1), 0.85 mg x kg(-1) and 90.63 mg x kg(-1) in Schima superba stand, and 28.50 g x kg(-1), 0.97 g x kg(-1), 0.23 g x kg(-1), 18.77 g x kg(-1), 73.40 mg x kg(-1), 1.45 mg x kg(-1) and 66.50 mg x kg(-1) in Castanopsis fissa stand, respectively. Soil bacteria accounted for > 94% of soil microbes, and their individuals were 41 x 105, 34 x 104 and 5.3 x 104 g(-1) in Michelia macclurei, Schima superba and Castanopsis fissa stands, respectively. The activities of soil urease, catalase and phosphatase in Castanopsis fissa stand were the greatest among the three stands, while soil cellulase activity in Michelia macclurei stand was greater than that in other two stands. In a word, soil fertility of Castanopsis fissa stand was the highest among three test stands. PMID- 16355773 TI - [Evapotranspiration of natural Quercus litaotungensis and Tilia paucicostata secondary stands in Liupan Mountains of Ningxia]. AB - With heat dissipation probe technique and combined with microlysimeter and hydrological methods, this paper studied the evapotranspiration of secondary Quercus liaotungensis and Tilia paucicostata stands, and its relationship to forest structure from August to September 2004. The results indicated that the stem sap flux density (SFD) of Quercus liaotungensis and Tilia paucicostata changed regularly from day to night in later growth season. In relatively still period (nighttime), the SFD kept low values continuously, usually below 0.05 microl x cm(-2). min(-1), while in active period (daytime), it increased quickly, usually below 0.25 microl x cm(-2) x min(-1). The daily transpiration of whole tree was estimated based on the calculation of daily cumulative SFD, which showed that there existed a significant difference between two tree species. The daily transpiration of Quercus liaotungensis reached 5.31 and 2.48 L x d(-1) in sunny- and cloudy-days, 2.35 and 3.75 folds as that of Tilia paucicostata, respectively. There was no significant difference in daily leaf transpiration rate between these two shrub species. During measurement periods, the average daily stand evapotranspiration was 1.45 mm x d(-1), including transpiration (0.72 mm x d( 1)), soil evaporation (0.19 mm x d(-1)), and canopy interception (0.54 mm x d( 1)), which accounted for 49.6%, 13.3% and 37.1% of the total evapotranspiration, respectively. The results showed that the difference of whole-tree transpiration between tree species contributed significantly to the difference of tree canopy transpiration, while the difference of shrub canopy transpiration was on account of the leaf amount in the canopy. The effects of vertical layers in the stand on total stand evapotranspiration also differed, with a percentage of 65.8%, 20.9% and 13.3% for tree canopy, shrub canopy and floor, respectively, which meant that the transpiration and interception of tree canopy contributed most to the total evpotranspiration, followed by shrub transpiration, soil evaporation, and grass transpiration. PMID- 16355774 TI - [Rainffall interception model of forest canopy: a preliminary study]. AB - Canopy interception is an important hydrological process in forest ecosystem, and its modelling is of significance to understand and estimate the rainfall interception by the canopy. In this paper, a canopy rainfall interception model was established by dividing a rain incident into a set of short period, calculating the rainfall distribution intercepted by the canopy, and educing the process of the rain incident. This model considered the effects of the dryness of canopy and trunk on the evaporation from wet canopy and trunk during one rain incident, and introduced two factors, leaf area index (LAI) and surface area of trunk per unit area of ground (SAI), when computing the evaporation. The application of the model to simulate the rainfall interception process in a Larix principis-rupprechtii plantation in Guyuan, south Ningxia Hui autonomous region of China showed that the simulated and measured throughfall were identical, and the absolute deviation between simulated and measured results was within +/- 1 mm. But, when the precipitation was smaller than 6 mm, the simulated throughfall was lower than the measured one. The values of simulated stemflow were lower than the measured values, and the relative deviation between simulated and measured values was smaller when the precipitation was larger. The process of throughfall in the forest was also simulated, with the results fitted well to the measured one. PMID- 16355775 TI - [Construction and verification of distributed rain-runoff model for forested watershed in alpine and gorge region]. AB - Considering of the main hydrological characteristics in the upper Zagunao River watershed of the upper reach of Minjiang River, general hydrological models were selected to construct the distributed rainfall-runoff process model for the forested watershed in typical alpine and gorge terrains. Calibration of too many regional parameters was avoided to assure the transportability of the models in similar watersheds. Two flow series of 1 000 hours in 1999 and 2000 were simulated by using the distributed model. The runoff hydrograph, accumulative runoff volume, peak discharge, and peak time showed good fittings with observed series. The simple structure and less empirical parameters gave the distributed model the ability to simulate the rainfall-runoff processes in similar watershed across scales, which provided the basic approach to the hydrological scaling research. PMID- 16355776 TI - [Application of wavelet transform to monthly runoff time serial analysis in Zagunao watershed, the upper Minjiang river]. AB - Runoff is an important component of regional water resources, and its dynamics is to some extent an indicator of water resources dynamics in a region. To know the runoff dynamics and water resources in a region is essential for the sustainable utilization and planning of water resources, and for the research on hydrological response of vegetation change at watershed scale. To disclose the water resources dynamics in Upper Minjiang River, one of the large reaches of Yangze River in southwest China, this paper analyzed the runoff dynamic features of Zagunao watershed, an important watershed in Minjiang River basin. Multi-Resolution Analysis (MRA) and periodicity analysis were conducted with Dmey wavelet function on the monthly runoff data from 1962 to 2002 observed by Zagunao hydrological station, which provided a data-based approximation on the evolution of monthly runoff in Zagunao watershed. It was found that the runoff dynamics in Zagunao watershed was relatively stable during 1962 - 1978, despite that this period was just in correspondence with the term of intensive deforestation activities in Upper Minjiang River basin. It was also clear that the runoff in Zagunao watershed was increased from 1986 to 1997, which was inconsistent with the commonly accepted viewpoint that the runoff decreased with increasing vegetation cover in forest watershed. However, the increasing trend from 1986 - 1997 was consistent with the research results in Yichang by Wang Wensheng and with the global warming at global and continent scale, which meant that global climate change plays a big role in runoff dynamics in Upper Minjiang River. Periodicity analysis showed that the rich-short water periodicity at the scale of 10 years (120 months) and 5 years (60 months) was 3 and 7 times, respectively, which could provide invaluable information for the eco-hydrological function research of forest landscape in Minjiang river basin and the sustainable utilization of water resources. The results from the case study highlighted that sometimes, it was very necessary to consider the role of global climatic warming in disclosing the hydrological response of land use/cover change at watershed scale, otherwise, it would be difficult to explain some hydrological phenomena. PMID- 16355777 TI - [Microclimate characters of urban forest in Shenyang]. AB - This paper analyzed the diurnal changes of the temperature and humidity in Shenyang urban forest during its different growth stages. The results showed that air temperature was the lowest in the morning and the highest at noon. The highest peak point of air temperature lagged behind that of solar radiation about 2 hours, due to the influence of forest environment. The annual air temperature amplitude was in order of stop-growth period (dormancy stage) (DS) > prophase of growth season (PGS) > anaphase of growth season (AGS) > medium-term of growth season (MGS). Soil temperature of PGS, MGS and AGS at daytime was higher than that at night, while that of DS was in reverse. The daily soil temperature amplitude at different growth stages was in order of upper soil layer (20 cm) >medium soil layer (40 cm) >lower soil layer(80 cm),while the diurnal soil temperature amplitude in the same soil layer was in order of PGS > AGS > DS > MGS (20 cm) and PGS > DS > AGS > MGS (80 cm). The diurnal change of relative humidity was reverse to that of air- and soil temperature, which was higher at night than at daytime, and was in order of MGS > AGS > DS > PGS. PMID- 16355778 TI - [Dynamics of total organic carbon (TOC) in hydrological processes in coniferous and broad-leaved mixed forest of Dinghushan]. AB - The total flux and concentration of total organic carbon (TOC) in hydrological processes in coniferous and broad-leaved mixed forest of Dinghushan were measured from July 2002 to July 2003. The results showed that the TOC input by precipitation was 41.80 kg x hm(-2) x yr(-1), while its output by surface runoff and groundwater (soil solution at 50 cm depth) was 17.54 and 1.80 kg x hm(-2) x yr(-1), respectively. The difference between input and output was 22.46 kg x hm( 2) x yr(-1), indicating that the ecosystem TOC was in positive balance. The monthly variation of TOC flux in hydrological processes was very similar to that in precipitation. The mean TOC concentration in precipitation was 3.64 mg x L( 1), while that in throughfall and stemflow increased 6.10 and 7.39 times after rain passed through the tree canopies and barks. The mean TOC concentration in surface runoff and in soil solution at 25 and 50 cm depths was 12.72, 7.905 and 3.06 mg x L(-1), respectively. The monthly TOC concentration in throughfall and stemflow had a similar changing tendency, showing an increase at the beginning of growth season (March), a decrease after September, and a little increase in December. The TOC concentration in runoff was much higher during high precipitation months. No obvious monthly variation was observed in soil solution TOC concentration (25 and 50 cm below the surface). Stemflow TOC concentration differed greatly between different tree species. The TOC concentration in precipitation, throughfall, and soil solution (25 and 50 cm depths) decreased with increasing precipitation, and no significant relationship existed between the TOC concentrations in stemflow, surface runoff and precipitation. The TOC concentrations in the hydrological processes fluctuated with precipitation intensity, except for that in stemflow and soil solutions. PMID- 16355779 TI - [Application of phenological pattern recognition in ecological dynamic forecasting]. AB - This paper described the principles, methods, and procedures of ecological dynamic forecasting by the automation techniques of pattern recognition and mathematical logic judgment on the basis of phenological data and model output maps from T42L9 numerical weather prediction model. This new forecasting method proposed on the basis of modern meteorology and automation techniques enables the classic phenology to apply to a new field ecological forecasting. It enables phenological forecasting to develop from single-station forecasting stage to regional forecasting stage, which is greatly corresponded to the development stage from single station forecasting stage to synoptic stage in weather forecasting, and enables agro-meteorological forecasting to develop from qualitative and statistical forecasting stage to ecological dynamic forecasting stage. With this new qualitative forecasting method, both the predicted objective and predictors are of considerable bio-physical interests. The ecological dynamic forecasting method could be applied to crop sowing, crop growth, irrigation and fertilization, and diseases and pests PMID- 16355780 TI - [Effects of continuous cropping on understorey species diversity in Eucalypt plantations]. AB - By the methods of time-space series analysis and field monitoring, this paper studied the species diversity and its maintaining mechanism in Dongmen Eucalypt plantations of Guangxi. The results indicated that during the period of 1998 - 2003, continuous cropping of Eucalyptus plantation resulted in a reduction of species diversity. The plant species number of second generation was reduced by 54.43% in a 667 m2 plot, and by 50% in 18 sampling plots (each of 4 m2), and the species richness and Shannon-Wiener index of 6-year second generation forest decreased by 39.39% and 17.76%, respectively, compared with the first generation. The continuous cropping not only decreased the species diversity, but also changed the species composition and characteristics of the community. Life-form spectra of Eucalypt plantations varied with generation. The plant life-form consisted of microphanerophytes, liana phanerophytes, and nana-phanerophytes in the first generation, while the herbaceous-phanerophytes and chamaephytes occurred in the second generation. The k-strategy and r-strategy species accounted for 69.32% and 30.68% in the first generation, while 47.83% and 52.17% in the second generation, respectively. Therefore, continuous cropping of Eucalypt plantations was advantage for herb, therophytes and r-strategy species, but disadvantage for woody plants and k-strategy species. Seed band and vegetative body in soil were considered to play an important role in maintaining diversity. A new hypothesis, initial plant Diaspora, on the maintaining mechanism of species diversity in Eucalypt plantation was put forward. PMID- 16355781 TI - [Effects of islanding on plant species diversity in Thousand-island Lake region]. AB - In this paper, the typical area of fragmentation in Thousand-island Lake region was selected to study the effects of islanding on plant species diversity after reservoir built. 26 quadrates were installed on 18 small, medium and large islands to investigate the species and number of trees and shrubs, with inland as the control One-way variance analysis (ANOVA) showed that the species richness of trees was significantly different (F = 13.055, P = 0.000) among all kinds of islands, which was significantly higher on large islands than on small and medium ones, but not significantly different from that on inland. The species richness of shrubs was not significantly different among all kinds of islands and inland. Spearman correlation analysis showed that the species richness of trees was significantly positively correlated with island area, while the correlation between specie richness of shrubs and island area was not significant. Shannon Wiener Index (H) analysis suggested that the diversity of both trees and shrubs on large islands was the highest, followed by on inland, but the diversity of shrubs was larger on small than on medium islands. The analysis of Simpson index and Pielou index showed that the species evenness of trees was the highest, but the dominance was the lowest on medium and large islands, while the species evenness of shrubs was the highest on medium and small islands, but the dominance was the lowest on small islands. PMID- 16355782 TI - [Secondary succession characteristics of vegetations on reclaimed land inside Chongming wetland seawall]. AB - National reserve Chongming Dongtan wetland was designated as an important international wetland (ramsar site) in 2002. The area outside 98 seawalls is the core area of the reserve, while their inside is the experimental area. A wetland restoration project was started in June 2003, and a large pond behind the seawall was unwatered, which changed the soil condition and appeared drought and salinization. The vegetations presented typical secondary succession, and Aeluropus littoralis and Suaeda glauca invaded the area originally dominated by reed. From July 2003 to April 2004, the soil moisture content and salinity of 15 samples were determined, with the biomass, height, density, and coverage of each kind of vegetations measured. The results showed that the vegetation layout had become an obviously zonary distribution of Aeluropus littoralis-Aeluropus littoralis/Suaeda glauca-Suaeda glauca/Phragmites communis-Phragmites communis community. Suaeda glauca gradually extended to the reed area, and became the dominant species of the area, with an average biomass of 2003 415.4 g x m(-2) in autumn, and of 2004 391.53 g x m(-2) in spring. The biomass and height of Suaeda glauca increased, and its density was getting stable, suggesting that the plant had strong growth ability in drought area. PMID- 16355783 TI - [Physiological responses of 2-year-old Acer davidii seedings to short-term enhanced UV-B radiation]. AB - At the Maoxian Ecological Experimental Station of Chinese Academy of Sciences in northwest Sichuan Province, 2-year-old native maple(Acer davidii) seedlings were potted outdoors with enhanced UV-B radiation(280 - 320 nm) of 0.27 W x m(-2) (7.7 kJ x m(-2) x d(-1)), which was approximated to the predicted enhanced UV-B reaching the earth surface when stratosphere ozone was depleted by 15% in the local area, with the control plant received ambient UV-B. The gas exchange index and chlorophyll fluorescence, and the contents of chlorophyll and UV-absorbing compounds were examined after 50 days of the radiation. The results showed that enhanced UV-B radiation significantly lowered the maximal net photosynthetic rate (CK = 6.214, TR = 4.452), raised the dark respiration rate(CK = 0.413, TR = 1.295) and light compensation point (CK = 21.629, TR = 59.861), but had little effect on quantum yield (CK = 0.021, TR = 0.032). Under enhanced UV-B radiation, the diurnal changes in net photosynthetic rate, water use efficiency, quantum efficiency of photosystem II centers (Fv/Fm), and quantum yield of photosystem II photochemistry (phi(pspi)) were reduced, chlorophyll a, total chlorophylls, and chlorophyll a/b (CK= 16.23, 5.39, TR = 13.17, 4.93) were also markedly reduced, but chlorophyll b remained nearly unchanged. Contrary to the previous studies, enhanced UV-B radiation decreased the content of UV-absorbing compounds (CK = 0.87, TR = 0.79) in 2-year-old Acer davidii seedling leaves, indicating that the measurement of leaf UV-B absorbing compounds didn't necessarily provide a good indicator of plant tolerance to UV-B. It could be concluded that enhanced UV-B radiation had some inhibitory effects on the photosynthesis of Acer davidii seedlings. Long-term researches are necessary to confirm this conclusion. PMID- 16355784 TI - [Fitness analysis of seed- and vegetative reproduction of clonal tree Symplocos laurina]. AB - There are two ways in Symplocos laurina propagation, clonal and sexual reproduction. The study showed that under different habitat conditions, Symplocos laurina could adopt different ways to propagate and occupy space. In conditions with abundant water and nutrient resources, such as in evergreen broad-leaved forests or bamboo forests, the survival rate and space-occupying ability of both ramets and sexual seedlings were relatively high, with clonal ramets took advantage in terms of number and space, suggesting that clonal propagation was the dominant way in such environments. Oppositely, in habitats lack of sufficient nutrition, the survival rate and space-occupying ability of seedlings were low, and grown-up plantlets would preempt in number and space occupation. Bottleneck in sexual propagation appeared in the stage from seed to seedling, while clonal propagation appeared during the period from seedling to ramet. The way of Symplocos laurina invasion was to settle a plantlet, and then occupied the space rapidly by clonal growth, with clonal seedlings dominated in initial stage and lost the advantage after 15 ages. PMID- 16355785 TI - [Landscape stability and its pattern transition in Jinghe watershed]. AB - Based on remote sensing and GIS, this paper analyzed the landscape change patterns in the Jinghe watershed of northwest Loess Plateau, one of the areas most suffered from soil and water erosion. The results indicated that the landscape structure in the watershed did not change very much over the past 15 years, and was dominated by grassland and arable land, which occupied 85% of total watershed area. Forest and other land use accounted for 10% and 5% of total area, respectively. A transition of landscape patterns occurred at large-scale watershed level. During 1986- 2000, the inter-transitions between different landscape types were very complicated, mainly occurred among woodland, grassland and arable land. The self-stableness of dominant landscape patterns was > 89%, and only 11% of them transferred into other types. Among landscape types, the one sharing small percentage of the total had a high speed of vanishing or transferring into other types. A significant transition was found between sub patterns of a large landscape pattern, e. g., the transition of medium- into low coverage grassland during 1986 - 1995 and the reverse during 1995 - 2000 was occurred, implying a situation of grassland degradation and improvement over the decades. The spatial-temporal changes of landscape features were observed in the watershed. In terms of spatial change, the dominance index and contagion index of the landscape showed an increasing trend, fractal dimension index showed a decreasing trend, while patch density showed an increasing trend, followed by a decreasing trend from upper to down streams. For temporal change, both landscape dominance and contagion decreased to a certain extent, fractal index remained stable, while patch density increased, showing a fragmentation direction of the landscape in the watershed. The NDVI value changed since the implementation of the policy of conversing farmland into grassland and forest in 2000, which was from 7.4% to 0.8% in low value class, from 29.8% to 25.1% in high value class, from 18.3% to 25.7% in higher value class, and from 2.3% to 5.5% in the highest class, showing an improving trend in landscape quality. PMID- 16355786 TI - [Landscape pattern and its functioning after ecological reconstruction in black soil of northern China]. AB - Taken Baiquan County in Heilongjiang Province as a case, this paper studied the effects of regional ecological reconstruction on agricultural landscape pattern and its functioning in soil and water conservation, with the spatial expression and quantity identification investigated based on Geographic Information System (GIS) and field data. The landscape pattern index in class scale, transition matrix of landscape elements, macrostructure of shelter-belt networks, and spatial distribution and hydrological characteristic of reservoirs and ponds in 1989 and 2002, as well as the effects of landscape pattern and its spatial configuration change on soil and water loss were compared. The results showed that artificial ecological reconstruction changed the quantity and spatial configuration of landscape patterns in regional scale, and exerted a significant effect on soil and water conservation. From 1989 to 2002, farmland area reduced 24 731.01 hm2, while forest land, grassland and water area increased 11 728.56, 1838.97 and 1190.97 hm2, respectively. The controlled eroded land increased by 55%, and the shelterbelt-protected farmland increased by 82.2%. The number and storage capacity of reservoirs and ponds added up to 1 490 and 2.4 x 10(8) m3, respectively, and 43.7% of intercepted surface runoff of whole landscape was collected. PMID- 16355787 TI - [Spatial pattern of sub-alpine forest restoration in west Sichuan]. AB - West Sichuan sub-alpine is an extension of Qinghai-Tibet Plateau to southeast China, which is covered mainly with dark coniferous forest. As a result of long term large scale over-logging, the forests have been greatly reduced and degraded. Nowadays, the forest restoration and regeneration in the region are being highlighted. Selecting Miyaluo as a case study area and employing the methods of plot investigation, ETM image interpretation, and overlaying vegetation map with digital topography, this paper analyzed the relations between the appearance and origin of four forest vegetation types, along with their topography differentiation and spatial patterns after a large scale logging and regeneration. The results showed that the appearance of forest vegetations was significantly correlated with their origin. Old coniferous forests (OC) were primitive ones, middle-aged and young coniferous forests (MYC) were from artificial regeneration, deciduous broadleaf forests (DB) were natural secondary ones, while mixed coniferous and deciduous forests (MCD) were partly from natural secondary ones and others from the conjunct action of artificial and natural regeneration. The main cut area in Miyaluo located in the sites with elevation from 2 800 to 3 600 m, where forest restoration appeared difference among different aspects. MYC was mainly distributed on sunny and half-sunny slope, DB and MCD were distributed on shady and half-shady slope, and OC were reserved on the sites with elevation more than 3 600 m. In the process of forest restoration, the four forest vegetation types were in mosaic pattern, and the landscape was seriously fragmentized. PMID- 16355788 TI - [Forest landscape restoration and its affecting factors in burned area of northern Great Xing'an Mountains--taking forest coverage as an example]. AB - Forest landscape restoration has been an attractive issue since the catastrophic fire took place on the northern slope of Great Xing'an Mountains in 1987. Based on the China forest inventory data and employing Kendall Bivariate and Distances Correlation Analyses, an investigation was made to search for what changes of the forest coverage pattern being happened in this area during the past 13 years after fire, and how the fire severity, foster type and terrain factors influenced the restoration of forest coverage. The results showed that the forest coverage in 2000 changed a lot, in comparing with that in 1987 before fire. The percentage of non-stocked land area and coverage grade declined markedly, with lower coverage grade increased. Among all test factors, fire severity which was inversely correlated with forest coverage grade was the key one. Though the regeneration measures didn't markedly affect forest coverage restoration within a short period, they might shorten the cycle of forest succession and promote the productivity of coniferous forest in the future. Among three terrain factors, slope was the strongest one affecting forest coverage, followed by position and aspect. PMID- 16355789 TI - [Relationships between soil and water loss and landscape pattern on Loess Plateau]. AB - Based on the theories of detrended canonical correspondence analysis (DCCA), 12 soil and water loss indices ("species") and 7 landscape indices ("environmental factors") were adopted to quantitatively analyze the effects of landscape pattern on the soil and water loss in Jinghe River basin on Loess Plateau. The results showed that the first four DCCA axes were significantly correlated to agricultural land ratio, landscape diversity index, forest land ratio, and landscape contagion index. The characteristics of soil and water loss changed obviously with landscape pattern gradient, e. g., Sanshuihe watershed with a forest cover larger than 65% and low landscape diversity had a big and stable runoff, but small sediment and low river sand content. The runoff generation coefficient and runoff depth as well as the sediment transport and river sand content increased with the increasing agricultural land ratio and landscape diversity and the decreasing forest land ratio. As for Honghe watershed with a very low forest cover but high agricultural land ratio (53.41%), and with a relatively high landscape diversity and complex landscape pattern, it had the highest river sand content and the biggest sediment transport, and its monthly runoff and sediment transport varied markedly. The soil and water loss in the Upper-Huan, Down-Huan and Dongchuan watersheds with smaller forest and agricultural land ratio and relatively simple landscape pattern was smaller than that of the watersheds with dominated agricultural land. PMID- 16355790 TI - [Landscape ecological planning of Jiaozuo city]. AB - With the application of the principles of landscape ecology and the techniques of geographic information systems, the landscape of Jiaozuo city was divided into six types, i. e., farmland, garden plot, woodland, residential and industrial area, waters, and others. The landscape diversity, dominance, fragmentation, and isolation indexes were calculated by models and GIS to analyze the spatial patterns of these landscape types. Based on the optimization of landscape spatial patterns, a pattern for developing a rational and natural ecosystem in Jiaozuo city was proposed, i. e., keeping four or five large natural patches, planning small artificial green patches, linking the large and small patches by various corridors to ensure the ecosystem inside the city to circulate well, combining residential areas properly to form a urban troop with Jiaozuo town as the center and with seven surrounding towns joined through highways to form a network city pattern. After doing these, urban built-up areas could be linked each other, and inlayed in a natural ecological matrix. This pattern could strengthen the ecological connection, raise the stability of the ecosystem, and maintain the balance of urban ecological system. PMID- 16355791 TI - [Landscape change in Kangbao County of Hebei {rovince]. AB - Based on RS image TM5 of 1999 and SPOT5 of 2003, this paper studied the landscape change from 1999 to 2003 of Kangbao County, Hebei Province. Logistic regression was adopted to analyze the driving factors, and Kappa index was used to evaluate the accuracy of the landscape classification result, which was 86.72% for 1999, and 89.76% for 2003. The area of cropland in Kangbao County reduced largely, while that of vegetable field, forestland and artificial grassland increased sharply, among which, vegetable field and artificial grassland had the greatest increasing rate. The landscape fragmentation aggravated in the region. The landscape change was mainly caused by the policy of reducing cultivated land, with the main driving factor slope, and the direct driving factor water-heat condition and soil fertility determined by slope aspect. PMID- 16355792 TI - [Fine-root character and its action mechanism of forest at its initial reestablished stage on degraded red soil]. AB - This paper studied the fine-root character and its action mechanism of forest reestablished by different models for 10 years on a seriously degraded red soil, and analyzed their correlation with soil property. The results showed that fine roots mostly distributed in 0-20 cm soil layer, accounted for 73.39% -87.41% of those in 0-40 cm soil layer. There were notable differences in nutrient storage of fine-roots under different reestablishment models. The fine- roots of medium density pure Pinus serotina forest had a much higher storage of total nitrogen, phosphorus and potassium, with the content of total nitrogen, phosphorus, potassium, calcium and magnesium being 84.47, 5.55, 38.42, 17.00 and 10.76 kg hm( 2), respectively. Soil organic matter, total nitrogen, available nitrogen, available potassium and pH had significant correlations with fine-root biomass (P < 0.05). The effects of fine-roots on soil physical properties mainly reflected on soil capillary porosity and total porosity. Fine-roots obviously increased the contents of > 0.25 mm and > 5 mm soil waterstable aggregates, and improved soil structure-stability. The correlations between fine-roots and soil bacteria and microbial quantities were significant. PMID- 16355793 TI - [Effects of different cutting intensities on above and underground growth of Stylosanthes guianensis]. AB - The study on the effects of different cutting intensities on the eco physiological characteristics and growth status of Stylosanthes guianensis showed that moderate cutting could promote the regrowth capacity of the grass. The total content of leaf chlorophyll had no significant difference between zero-, light and heavy cutting, but the ratio of leaf chlorophyll a/b increased to 1.59: 1 and 1.52: 1 under light-and heavy cutting, respectively, compared with 1.22:1 under zero-cutting, which could be very helpful for the plant to over-compensate itself. Cutting also affected the growth status and activities of plant roots. At the earlier stage of heavy cutting, the total length, total surface area and mean diameter of plant roots decreased by 54.9%, 66.5% and 27.2%, respectively, and root activities also decreased greatly, in comparing with zero-cutting. But, the above-mentioned indices could be recovered to a higher level under different cutting treatments than under zero-cutting at the later growth period of the grass. As for the total annual yield of the grass, it was 3 179.8 g x m(-2) under light-cutting, 3006.1 g x m(-2) under heavy- cutting, and 2 936.98 g x m(-2) under zero-cutting, indicating that rational cutting could improve grass productivity. PMID- 16355794 TI - [Economic values of nitrogen transformation in rice field ecosystems]. AB - Based on the 2002 field experimental results in the Wusi Farm of Fengxian County in Shanghai and related studies, and by the methods of ecological economy and environmental economy, this article examined and estimated the nitrogen input and output, and the economic values of nitrogen transformation in rice field ecosystems. The results indicated that the plot without N fertilization showed a deficit of nitrogen budget, while those with urea application showed a surplus. The economic value of the plot without urea amendment was evaluated to be positive, while that of the plots with urea application was calculated to be negative. The negative value of the plots with urea amendment resulted from the nitrogen losses through ammonia volatilization, nitrogen leaching and runoff, which had serious harmful impacts on environment and human society. Therefore, only when some measures were taken to encourage and facilitate farmers to improve the efficiency of urea application and to mitigate the environment problems from N losses in the process of food production, the advantages of nitrogen transformation in rice fields could be promoted, and the sustainability of agriculture would become reality. PMID- 16355795 TI - [Sex pheromone secondary components of Indian meal moth Plodia interpunctella in China. HU wenlil 2, DU]. AB - Indian meal moth Plodia interpunctella Hubner (Lepidoptera: Pyralidae) is considered as an important insect pest infesting stored grains and other products in China. The major sex pheromone component of P. interpunctella, Z9, E12-14: OAc (TDA), has already been identified. Though the efficiency of male capture by using the bait with this component alone is quite good, the pheromone system is far from fully understood. The identification with capillary chromatographic analysis and GC-MS methods showed that there were four main components, i. e., Z9, E12-14: OAc(A), Z9, E12-14: OH (B), Z9, E12-14: Ald(C), and Z9-14: OAc(D), in the sex pheromone gland of female P. interpunctella, and the ratio of these four components was A: B: C:D= 100:22: 12:9. Wind tunnel experimental results suggested that the response of male P. interpunctella to a blend (A: B: C: D = 8:2:1:0.8) was not significantly different from that to female sex pheromone gland extracts. PMID- 16355796 TI - [Phosphate adsorption and desorption characteristics of several fly ashes]. AB - Through adsorption-desorption experiment and incubation test, this paper studied the phosphate adsorption and desorption characteristics of several fly ashes. The test fly ashes contained 0.545 - 4.540 g x kg(-1) of total P and 19.55 - 163.0 mg x kg(-1) of available P, which were significantly higher than those in soils. The P adsorption capacity of fly ashes increased with increasing added P, while their P adsorption rate was in adverse. Fly ashes had a higher P adsorption rate but a lower P desorption rate than soils, mainly because fly ashes had more P adsorption sites and stronger bound energy. In this study, Langmuir, Freundlich, and Temkin equations were fit to the measured data, and the MBC value in Langmuir equation, a value in Freundlich equation, and k2 value in Temkin equation could be used as a comprehensive index to characterize the potential phosphate adsorptivity of fly ashes. The larger these values were, the stronger the P adsorptivity was. The MBC, a and k2 value of 5 fly ashes collected from different locations was in order of Xiangtan power factory (5 167.7, 4 056.2 and 831.5) > Yueyang paper factory (1 650.7, 2803.4 and 711.9) >Huaneng power factory (303.0, 1677.6 and 368.7) > Zhuzhou power factory (34.7,413.48 and 213.8) > Dongting nitrogen fertilizer factory (34.7, 413.48 and 213.8). The P fixation by fly ashes generally increased with their increasing water content, and their phosphate adsorption was mainly specific adsorption and chemical precipitation, suggesting that the P fixation and the water content of fly ashes should be considered when fly ash was used as soil amendment or as compound fertilizer filling substance. PMID- 16355797 TI - [Degradation-detoxification behavior of methamidophos in phaiozem and burozem rhizosphere]. AB - With plastic rhizobox system, this paper studied the degradation behavior of methamidophos in the rhizosphere and non-rhizosphere of phaiozem and burozem planted with soybean. The results suggested that methamidophos could be rapidly decomposed in soil environment despite its higher acute toxicity, and its residual amount in phaiozem was generally below the level in burozem under same condition. In the absence of soybean (control), the residual amount of methamidophos at the 2nd day of incubation was about 33% in burozem, whereas only about 26% in phaiozem. An accelerated degradation of methamidophos in soybean rhizospheric soil was observed, especially in phaiozem. At the 9th day of incubation, the insecticide residue in the middle rhizobox phaiozem and burozem with soybean was decreased by 87.5% and 76.0%, respectively, compared with that of the control. The degradation process of methamidophos in soil environment followed the first-order equation, and its half-life was about 2 days. PMID- 16355798 TI - [Bt toxin distribution in transgenic Bt cotton and soil system]. AB - This study showed that the amounts of Bt toxin expressed in transgenic Bt cotton leaves and stems (103.5 - 134.1 ng x g(-1)) were rather higher than those expressed in transgenic Bt cotton roots (44.7 - 21.2 ng x g(-1)), indicating that total amount of soil Bt toxin introduced by transgenic Bt cotton could be decreased through treating its above-ground biomass. In comparing with the amount of Bt toxin expressed in transgenic Bt cotton plant, that expressed in its root exudates was rather lower, showing that the effects of plant Bt toxin on soil ecosystem would be limited if other sources of Bt toxin introduced into soil were controlled. PMID- 16355799 TI - [Research advances in inverse methods used for modeling plant-atmosphere exchange]. AB - To estimate the source/sink and the vertical fluxes of mass and energy within and above plant canopies continues to be a critical research problem in biosphere atmosphere exchange processes. The underlying approaches in such problem are to exploit the natural properties of turbulence within and above vegetation, such as Lagrangian inverse analysis, high order Eulerian closure model, and hybrid Eulerian-Lagrangian method. This paper introduced the recent development in multilayer turbulent transport methods to compute the distributions of the strengths of scalar sources and sinks within plant-atmosphere continuum, and in particular, focused on the so-called "inverse methods", and described above three methods and their characteristics in detail. The limitation and prospect of these methods were also mentioned. PMID- 16355800 TI - [Research progress on unsaturated and saturated soil water movement in forest catchments]. AB - This paper reviewed the studies on the movement ways, i. e., infiltration, phreatic evaporation, ground water recharge and interflow, of unsaturated and saturated soil water in forest catchments, and introduced the present advances in soil hydraulic parameters, including soil water characteristic curve, and unsaturated and saturated soil hydraulic conductivity. Research directions in the future were also proposed. PMID- 16355801 TI - [Advances in research effects of temperature fluctuation on aquatic fauna]. AB - This paper reviewed the researches on the effects of diurnal temperature fluctuation on aquatic fauna, including the test species involved, design of thermal regimes, growth responses of aquatic fauna to various fluctuated temperature, and growth-promotion mechanisms of temperature fluctuation. The problems existed in related researches and the application perspectives in the future were also highlighted. PMID- 16355802 TI - [Understory vegetation composition and main woody population niche of artificial masson pine forest in south subtropical area]. AB - This paper studied the characteristics of the natural understory vegetation in the artificial masson pine forest replanted after Chinese fir cut for 13 years. The results indicated that there were 100 species of understory vegetation, including 74 species of shrub or small arbor, accounting for 74.0% of the total species, belonging to 41 faculties and 63 categories, and 26 herbs belonging to 14 faculties and 23 categories. The tropical and subtropical elements were dominant in the vegetation, while the temperate ones were scare. The species diversity and richness were decreased after masson pine regenerated from Chinese fir. The analysis of important value showed that the main species of shrub or small arbor were Tinomiscium tonkinensis, Mycetia longiflora, Evodia lepta, Paederia scandens, Mussaenda pubuscens, Actinodaphne pilosa and Quercus glauca, and those of herb were Cyrtococcum patens, Blechnunm orientale, Dicranopteris dichotoma, Adiantum flabellulatum, Adiantum edgeworthii and Pteris semipinnata. The biomass of the vegetation in different slope positions was decreased in order of upside> downside > middle part. There was a significantly positive correlation between the niche breath and the important value of main woody population. The important values of woody populations were higher than those of the others, and their niche breadths were also larger, which meant that they were more adaptable to the habitat. The niche overlaps of six population counterparts Tinomiscium tonkinensis-Mycetia longiflora, Tinomiscium tonkinensis-Evodia lepta, Mussaenda pubuscens-Maesa japonica, Quercus glauca-Aphananthe aspera, Actinodaphne pilosa Saraca chinensis and Evodia lepta-Quercus glaucathese were very large, which could offer information for the restoration and reconstruction in selecting natural vegetation. The large niche overlaps of major woody population counterparts suggested their similar utilization of natural resources. PMID- 16355803 TI - [Biodiversity of main coniferous forests at low elevation of Three Gorges Reservoir area]. AB - The study showed that at low elevation of Three Gorges Reservoir area, Pinus massoniana, Cupressus funebris and Cunninghamia lanceolata were the main coniferous forests, which were respectively divided into 14, 9 and 5 community types based on field investigation. The whole trend of their species richness index, species diversity index and species evenness index was that shrub layer and herb layer were larger than tree layer. The diversity indices had no obvious trend along the low elevation gradients, which was probably relevant to the high intensity of human activities in the area. PMID- 16355804 TI - [Application of fractal theory in studying soil aggregates under different skidding modes]. AB - In this paper, the characters of soil aggregates under different skidding modes were investigated by fractal theory. The results showed that the particle fraction of soil aggregates was smaller under tractor- than under animal traction skidding. As for different parts of skidding road, the particle fraction of air dried soil aggregates was in the order of bottom (2.309) > middle (2.291) > upper (2.014), while that of water-stable aggregates followed the sequence of bottom (2.04) < middle (2.143) < upper (2.262). The particle fraction of aggregates was 2.215 in forest soil and 2.175 in skidding roads. Skidding movement could improve soil aggregates when the cutting intensity was 15%. Tractor skidding was more favorable for soil aggregates formation. PMID- 16355805 TI - Leptin levels in boys with pubertal gynecomastia. AB - BACKGROUND: It has been reported that there is a relationship between circulating leptin and sex steroid hormones and leptin is able to stimulate estrogen secretion by increasing aromatase activity in adipose stromal cells and breast tissue. Leptin receptors have been also shown in mammary epithelial cells and it has been suggested that leptin is involved in the control of the proliferation of both normal and malignant breast cells. AIM: To investigate circulating leptin levels in boys with pubertal gynecomastia. METHODS: Twenty boys with pubertal gynecomastia who were in early puberty and had no obesity, and 20 healthy individuals matched for age, pubertal stage and body mass index (BMI) with the study group, were enrolled in the study. Body weight, height and left midarm circumference (MAC) and left arm triceps skinfold thickness (TSF) were measured and BMI was calculated. A fasting blood sample was collected and routine hormonal parameters including prolactin, beta-human chorionic gonadotropin (betaHCG), total and free testosterone, estradiol, luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin, androstenedione (AS) and dehydroepiandrosterone sulfate (DHEAS) levels were studied. Serum leptin levels were analyzed using radioimmunoassay. RESULTS: The mean ages of the study and control group were not different (13.9 +/- 0.89 and 14.2 +/- 0.66, respectively). No significant difference was found for BMI, MAC and TSF values between the two groups. There was no significant difference for hormonal parameters including FSH, LH, total and free testosterone, estradiol, AS, DHEAS and estradiol/total testosterone ratio between boys with pubertal gynecomastia and the controls. Serum leptin levels were found significantly higher in the study group compared with the healthy controls (5.58 +/- 0.81 and 2.39 +/- 0.29 ng/ml, respectively; p <0.001). No correlation could be determined between serum leptin levels and hormonal parameters. CONCLUSION: The presence of higher leptin levels in boys with pubertal gynecomastia indicates that leptin may be involved in the pathogenesis of pubertal gynecomastia. The role of circulating leptin in pubertal gynecomastia is probably related to increase in estrogen and/or estrogen/ androgen ratio by the stimulating effect of leptin on aromatase enzyme activity in both adipose and breast tissues, or a direct growth stimulating effect of leptin on mammary epithelial cells, or increase in sensitivity of breast epithelial cells to estrogen with inducing functional activation of estrogen receptors by leptin in breast tissue. PMID- 16355806 TI - Endocrine disorders associated with holoprosencephaly. AB - OBJECTIVE: To investigate the incidence of endocrinopathies in holoprosencephaly (HPE) and correlate the severity of the endocrinopathies with the neuroanatomic abnormalities. STUDY DESIGN: We reviewed the histories and medical records of 117 children with HPE for endocrinopathies and related treatments. Neuroimaging studies were graded for severity of HPE, hypothalamus non-separation, and pituitary abnormalities. RESULTS: Diabetes insipidus (DI) occurred in 70% of patients with classic HPE. The severity of the DI correlated with the grade of HPE and hypothalamic non-separation (p < 0.0001). Anterior pituitary dysfunctions were much less common. Hypothyroidism was identified in 11% of patients, hypocorticism in 7%, and growth hormone deficiency in 5%. Only one patient with middle interhemispheric variant of holoprosencephaly (MIH) had any of these disorders. CONCLUSIONS: Patients with HPE have a high incidence of DI that may be related to the failure of cleavage of hypothalamic nuclei. Anterior pituitary dysfunctions are much less common than DI. PMID- 16355807 TI - Are factors at diagnosis of growth hormone deficiency in childhood associated with persistence of growth hormone deficiency into adult life? AB - A proportion of children with growth hormone deficiency (GHD) have persistence of GHD as young adults. To date, no markers have been shown in childhood to have predictive value in determining persistence of GHD into adult life. We examined the hypothesis in 31 patients that variables present at the time of diagnosis of childhood-onset GHD, or those related to the early response to growth hormone (GH) therapy, are associated with the likelihood of persistence of GHD. The results show that, as previously demonstrated, children with GHD are more likely to have persistent severe GHD in adult life when the diagnosis is associated with other pituitary hormone deficiencies (p = 0.02), abnormal pituitary neuroimaging (p = 0.003), induced puberty (p = 0.001), early age of diagnosis (p = 0.03) and lower peak GH response at the first dynamic GH test in childhood (p = 0.02). However, there are no associations of persistent severe GHD with the pattern of pretreatment growth or growth response to GH treatment in the initial phase. PMID- 16355808 TI - Response to growth hormone with respect to pubertal status on increased dose in idiopathic growth hormone deficiency: an analysis of Turkish children in the KIGS database (Pfizer International Growth Study). AB - AIM: To compare the growth response to growth hormone (GH) treatment in patients with idiopathic GH deficiency (IGHD) who were prepubertal with the response of those who were pubertal at the onset of GH therapy on an increased GH dose. PATIENTS AND METHODS: Among the Turkish patients enrolled in the Pfizer International Growth Study (KIGS) database with the diagnosis of IGHD, the growth data over 2 years of GH therapy were analyzed longitudinally of 113 (79 M) prepubertal (Group 1) and 44 (33 M) pubertal (Group 2) patients. Pubertal signs were reported to be present initially or to have appeared within 6 months of GH therapy in Group 2. Mean +/- SD age at onset of therapy was 8.7 +/- 3.5 and 13.5 +/- 1.8 years; height SDS -4.2 +/- 1.4 and -3.2 +/- 1.1 (p < 0.05) in Groups 1 and 2, respectively. Mid-parental height (MPH) SDS did not show a significant difference between the two groups (-1.5 +/- 1.1 vs -1.7 +/- 1.1). RESULTS: Delta height SDS over 2 years of therapy was significantly higher in Group 1 (1.1 +/- 1.0) than in Group 2 (0.7 +/- 0.6) (p <0.05) in spite of a significantly lower dose of GH (14.6 +/- 3.3 in Group 1 vs 17.0 +/- 3.1 IU/m2/week in Group 2, p < 0.05). Ht--MPH SDS showed an increase from -2.4 +/- 1.7 to -1.4 +/- 1.5 in Group 1 and from -1.5 +/- 1.5 to -0.8 +/- 1.3 in Group 2. Overall delta height SDS showed negative correlations with age (r = -0.32), height SDS (r = -0.41) and height--MPH SDS (r = -0.40) at onset of therapy (p < 0.001). CONCLUSIONS: These data show that in IGHD the slight increase (15-20%) in the dose of GH during puberty was not adequate to maintain height velocity at the same magnitude as in prepuberty, and thus was not cost effective. PMID- 16355809 TI - Growth hormone releasing hormone receptor (GHRH-r) gene mutation in Indian children with familial isolated growth hormone deficiency: a study from western India. AB - BACKGROUND: Various mutations of the growth hormone releasing hormone receptor (GHRH-R) gene have been recently described to cause familial isolated growth hormone (GH) deficiency (FIGHD), with the GHRH-R nonsense mutation E72X reported in patients with FIGHD from South Asia. The molecular genetic basis of FIGHD in Indian children is not known. OBJECTIVE: To look for the GHRH-R E72X non-sense mutation in our patients with FIGHD and describe its clinical phenotype. PATIENTS AND METHOD: A total of 31 patients from 22 families diagnosed 4-20 years previously, 20 patients with familial IGHD-IB from 11 families and 11 patients with non-familial isolated GH deficiency (NFIGHD) (phenotypes IGHD-IB in eight patients and -IA in three) were included. Twenty-eight of 31 patients with IGHD IB came from two states of Western India, 27 of them Hindus from 18 families (three consanguineous) and one from an inbred Moslem kindred. RESULTS: Twenty-two of the patients (71%) (18 FIGHD and four NFIGHD) had a homozygous G-->T transversion in exon 3, with this GHRH-R gene mutation E72X in 90% (18/20) of patients with FIGHD, 36% (4/11) of NFIGHD, altogether 78% (22/28) with phenotype IB. One parent pair with IGHD had homozygous E72X mutation, the rest were heterozygous carriers. Two siblings with IGHD due to homozygous E72X mutation were also heterozygous carriers for GH-1 gene 6.7 kb deletion, inherited from their mother, heterozygous for both GH-1 and GHRH-R mutations. Initial chronological age was 10.89 +/- 3.69 years, bone age 6.4 +/- 3.4 years, and mean height SDS was -5.83 +/- 1.41. The clinical phenotype, with sharp features, lean habitus, lack of frontal bossing or hypoglycemia, was characteristic. The mean peak GH was 1.25 +/- 0.75 ng/ml, IGF-I and IGFBP-3 below -2 SDS with no response to GHRH in those tested. MRI (n = 10) showed pituitary hypoplasia, mean vertical height 2.61 +/- 0.76 mm. Among the other 7/11 NFIGHD patients, four with phenotype IB were negative for genotypes tested in this study; of three patients with phenotype IA, two had the GH-1 gene 6.7 kb deletion, and one was a compound heterozygote with 6.7 and 7.6 kb deletions. CONCLUSIONS: The majority of patients with FIGHD from different communities belonged to non-consanguineous Hindu families from Western India. The GHRH-R gene E72X mutation was found in 71% of this series, in 90% of FIGHD, 36% of NFIGHD, and in 78% with phenotype IB. The characteristic phenotype helped in suspecting this mutation. GHRH-R gene mutations may be the most reasonable candidate for IGHD-IB with the E72X mutation predominating in the Indian subcontinent. More extensive studies need to be undertaken. PMID- 16355810 TI - Body composition, dehydroepiandrosterone sulfate and leptin concentrations in girls approaching menarche. AB - There is strong evidence that the initiation of adrenarche and gonadarche during puberty in girls depends on body mass in general and body fat in particular. The aim of this study was to analyze changes in body composition, i.e. body fat (BF), fat percentage (BF%), and lean tissue mass (LM) in girls during pre-menarcheal stages of development, including the earliest stage lacking clinical manifestations of changes in primary, secondary, and tertiary sexual characteristics. Puberty was assessed according to clinical and ultrasonographic staging of sex features developed by us. Concentrations of leptin and DHEA-S were compared and related to changes in body composition. SUBJECTS AND METHODS: The study was carried out on 65 healthy girls aged 8 years and older who were followed every 3 months over a 5-year period. Age, height, weight, and BMI were recorded. Body composition (BF, BF%, LM) was determined with an infrared method. Tertiary sexual features were staged according to Tanner. Vaginal secretion was assessed according to Peter et al. Transabdominal ultrasound of the uterus and ovaries was performed with the bladder unvoided. Groups were formed according to developmental stage: E0A = pre-estrogenization (no ultrasonographic or clinical evidence of estrogenization); E0B pre-estrogenization with 'luminosity' of mucus in cervical canal; E1 = onset of estrogenization; E = full estrogenization; M = menarche +/- 3 months. Concentrations of DHEA-S and leptin were determined by radioimmunoassay. RESULTS: BF in prepubertal girls averaged 16%. At menarche, BF was 23.9%. Body weight at menarche was 50.6 kg and the LM/BF ratio was 3.0. High leptin concentrations were found in E0B and M groups. Leptin concentrations were lowest during full estrogenization (E). Positive correlations of leptin with BF and LM were found in girls during developmental stages preceding menarche. Mean concentration of DHEA-S started at 1,091.6 microg/l during E0A stage, dropped significantly on passing to E0B (p <0.05), and increased by menarche. DHEA-S levels were found to correlate with BF, LM, and leptin in E0A, E0B, and E1 groups. Correlation coefficients were highest (DHEA-S/BF r = 0.61; DHEA-S/LM r = 0.54; DHEA-S/LEP r = 0.57) in the E0A group, i.e. about 5 months before the appearance of 'luminosity' of cervical mucus, considered to be the first ultrasonographic sign of puberty. Apparently, leptin stimulates somatic maturation during this stage of gonadarche which terminates with menarche. The action of DHEA-S is exerted during the early stages of female puberty. PMID- 16355811 TI - Valproate-induced insulin resistance in prepubertal girls with epilepsy. AB - Valproate is commonly used for treatment of a variety of seizure types in both children and adults. However, if the medication is started before the age of 20 years, it may affect reproductive endocrine functions. In order to investigate the possible role of valproate treatment in the development of obesity, hyper insulinism and polycystic ovaries, we studied metabolic parameters and ovarian morphology/size in prepubertal girls with epilepsy. Our study included 14 girls with epilepsy and 15 healthy age-matched controls. The age of the patients ranged from 7 years to 13 years. Mean body weight, fasting serum insulin and glucose levels and HOMA index of girls in the study group were significantly greater than those of the control girls (p < 0.05). Serum androstenedione, prolactin and free testosterone were significantly lower in the VPA-treated girls than in the controls, whereas SHBG level was higher (p < 0.05). There was no difference between the groups for ovarian morphology. In conclusion, our findings showed that valproate treatment may lead to hyperinsulinemia and hypoandrogenism during the prepubertal period. This emphasizes that a mature adult endocrine system may not be necessary for the development of VPA-related hyperinsulinemia. PMID- 16355812 TI - Novel deletion mutations of the DAX1 (NR0B1) gene in two Taiwanese families with X-linked adrenal hypoplasia congenita. AB - OBJECTIVE: To analyze the DAX1 (NROB1) gene in Taiwanese families with adrenal hypoplasia congenita. PATIENTS AND METHODS: Two unrelated Taiwanese patients were followed up at our pediatric endocrine clinic. Both patients presented with adrenal crisis. One patient entered puberty spontaneously. However, arrest of puberty was noted in the following years and hypogonadotropic hypogonadism was confirmed by GnRH test. The other patient was still prepubertal. Sequencing of the DAX1 (NROB1) gene was carried out in both patients and their respective family members. RESULTS: Two different novel mutations were identified. The first patient had one base (G) deletion at nucleotide 159, resulting in a frame-shift and a premature stop codon at position 84. The other patient had one base (G) deletion at nucleotide 831, leading to a frameshift and a premature stop codon at position 371. Family studies revealed that their mothers and sisters were heterozygotes for the mutations while their maternal grandmothers did not carry the mutations. CONCLUSIONS: Two Taiwanese patients with adrenal hypoplasia congenita were detected to have novel mutations of the DAX1 (NR0B1) gene. Family studies suggested that such mutations resulted from de novo mutation of the DAX1 (NROB1) gene in their mothers. These data indicate that molecular analysis of the DAX1 (NR0B1) gene is important for the diagnosis and genetic counseling of children with primary adrenal insufficiency. PMID- 16355813 TI - Comparison of clinical, radiological and molecular findings in Korean infants and children with achondroplasia and hypochondroplasia. AB - Achondroplasia (ACH) and hypochondroplasia (HCH) share clinical features characterized by disproportionate short stature with rhizomelic shortening of the limbs. In an attempt to clarify genotype-phenotype correlation in ACH and HCH, we investigated the presence of the previously identified mutations of FGFR3 in 26 patients with ACH- or HCH-mimicking features and compared clinical and radiographic findings between the two groups. Using genomic DNA sequencing and RFLP analysis, G380R, an ACH-specific mutation, and N540K, an HCH-specific mutation, were detected in 13 patients (50%) and five patients (19%), respectively. No mutations of FGFR3 were detected in eight patients (31%). No remarkable clinical or radiological differences were evident among the ACH infants and children with G380R, the HCH patients with N540K, and the patients without verified mutations. These results suggest that genotype-based diagnosis needs to precede proper genetic counseling for patients with ACH or HCH, which show very similar clinical and radiological features. PMID- 16355814 TI - Serum zinc, insulin-like growth factor-I and insulin-like growth factor binding protein-3 levels in children with type 1 diabetes mellitus. AB - BACKGROUND: Growth is impaired during the course of diabetes mellitus (DM). Derangement of the growth hormone/insulin-like growth factor (IGF) axis, insulinopenia and zinc deficiency are the possible causative factors of this impairment. Zn supplementation is proven to attenuate hyperglycemia in mice but its use to ameliorate impaired height is still a matter of discussion. OBJECTIVE: To investigate serum Zn, IGF-I and IGF binding protein-3 (IGFBP-3) levels and to emphasize the potential beneficial effects of Zn supplementation for the prevention of growth failure in children with type 1 DM (DM1). PATIENTS AND METHODS: Twenty-eight patients with DM1 and 15 control children were included in the study. Zn levels were measured by flame atomic absorption spectrophotometry; IGF-I and IGFBP-3 levels were measured by immunoradiometric assay. RESULTS: Mean serum Zn levels were significantly lower in diabetic children taken as a whole and as their pubertal subgroup compared to the controls. Mean serum IGF-I and IGFBP-3 levels were significantly lower in both prepubertal and pubertal diabetic groups compared to those of control groups. CONCLUSION: From the results of our study, it can be hypothesized that serum Zn levels should be closely monitored during the course of DM1 and supplementation may be given to patients, especially at the time of puberty. This hypothesis needs to be confirmed by further studies. PMID- 16355815 TI - Primary ovarian failure and deletions of the long arm of chromosome 3. AB - A 14 year-old girl was found to have a deletion of the distal segment of chromosome 3 [46,XX,Del(3)(q28-29)]. The main features of this presentation were mild intellectual disability, facial dysmorphism, short stature, kypho-scoliosis, and primary ovarian failure, an association that has not been described before in association with chromosome 3 deletion. The phenotype and presentation are compared with those of previous case reports. PMID- 16355816 TI - Hypertrichosis cubiti (hairy elbow syndrome): a clue to a malformation syndrome. AB - Hypertrichosis cubiti (HC) or hairy elbow syndrome (OMIM # 139600) consists of a localised form of long vellus hair on the extensor surfaces of the distal third of the upper arm and the proximal third of the forearm bilaterally, or occasionally on other parts of the body. In the 28 cases reported in the literature so far the elbow hair abnormality was either isolated or associated with short stature or other physical abnormalities. Most of these cases were sporadic, but autosomal dominant as well as autosomal recessive inheritance patterns have been postulated. We report on three unrelated girls (aged 7 to 11 years) of whom one presented with excess hair in the elbows alone and the other two had associated abnormalities including short stature, dysmorphic facial features and mental retardation. The literature on this subject has been reviewed and the authors focus on cases of HC with associated anomalies. A pathogenic explanation by somatic mosaicism is proposed. PMID- 16355818 TI - Sustained decrease in the rate of Escherichia coli O157:H7-positive raw ground beef samples tested by the Food Safety and Inspection Service. PMID- 16355817 TI - Central precocious puberty with an incidental suprasellar lipoma. PMID- 16355819 TI - Internalization of Escherichia coli O157:H7 following biological and mechanical disruption of growing spinach plants. AB - The internalization and persistence of a bioluminescent Escherichia coli O157:H7 Ph1 was investigated in growing spinach plants that had been either biologically or mechanically damaged. In control (undamaged) plants cultivated in soil microcosms inoculated with E. coli O157:H7 Phl, the bacterium was recovered from surface-sterilized root tissue but not from leaves. Mechanical disruption of the seminal root and root hairs of the plants did not result in the internalization of the pathogen into the aerial leaf tissue. When imprints of the root tissue were made on plates of tryptic soy agar plus ampicillin, no colonies of E. coli O157:H7 were observed around damaged tissue. The roots of growing plants were exposed to the northern root-knot nematode, Meloidogyne hapla, in the presence of E. coli O157:H7. Although this treatment caused knot formation on the roots, it did not enhance the internalization of the bacterium into the plant vascular system. Coinoculation of intact leaves with E. coli O157:H7 and the phytopathogen Pseudomonas syringae DC3000 resulted in localized necrosis, but the persistence of the human pathogen was not affected. The mechanical disruption of roots does not result in the internalization of E. coli O157:H7 into the aerial tissue of spinach, and there does not appear to be any effect of P. syringae in terms of enhancing the persistence of E. coli O157:H7 in spinach leaves. PMID- 16355820 TI - Spatial distribution of Salmonella, Escherichia coli O157:H7, and other bacterial populations in commercial and laboratory-scale sprouting mung bean beds. AB - The reliability of testing spent irrigation water to assess the microbiological status of sprouting mung bean beds has been investigated. In commercial trials, the distribution of opportunistic contaminants within 32 bean sprout beds (25 kg of mung beans per bin) was assessed 48 h after germination. The prevalence of generic Escherichia coli, thermotolerant coliforms, and Aeromonas in sprouts (n = 288) was 5, 11, and 39%, respectively, and 57, 70, and 79% in the corresponding spent irrigation water samples (n = 96). Contamination was heterogeneously distributed within the seedbed. In laboratory trials, beans inoculated with a five-strain cocktail of either Salmonella or E. coli O157:H7 (10(3) to 10(4) CFU/g) were introduced (1 g/500 g of noninoculated seeds) at defined locations (top, middle, or base), and the beans were then sprouted for 48 h. When seeds inoculated with pathogens were introduced at the base or top of the seedbed, the pathogens were typically restricted to these sites and resulted in 44% of the spent irrigation water samples returning false-negative results. Introducing inoculated beans into the middle or at the presoak stage enhanced the distribution of both pathogens within the subsequent sprout bed and resulted in comparable levels recovered in spent irrigation water. The study demonstrated that even though screening a single sample of spent irrigation water is more reliable than testing sprouts directly, it does not provide an accurate assessment of the microbiological status of sprouting mung bean beds. Such limitations may be addressed by ensuring that bean batches are mixed prior to use and by taking spent irrigation water samples from multiple sites at the latter stages of the sprouting process. PMID- 16355822 TI - Salmonella and the sanitary quality of aquacultured shrimp. AB - In this study, we examined the prevalence of Salmonella and coliform bacteria on shrimp aquaculture farms to develop guidelines or preventative measures for reducing Salmonella and fecal contamination on products harvested from these farms. The U.S. Food and Drug Administration, in conjunction with foreign government regulatory agencies, the aquaculture industry, and academia affiliates, analyzed 1,234 samples from 103 shrimp aquaculture farms representing six countries between July 2001 and June 2003 for fecal coliforms, Escherichia coli, and Salmonella. A significant relationship was found (P = 0.0342) between the log number of fecal bacteria and the probability that any given sample would contain Salmonella. The likelihood of any given sample containing Salmonella was increased by 1.2 times with each 10-fold increase in either fecal coliform or E. coli concentration. The statistical relationship between Salmonella concentration and that of both fecal coliforms and E. coli was highest in grow-out pond water (P = 0.0042 for fecal coliforms and P = 0.0021 for E. coli). The likelihood of finding Salmonella in grow-out pond water increased 2.7 times with each log unit increase in fecal coliform concentration and 3.0 times with each log unit increase in E. coli concentration. Salmonella is not part of the natural flora of the shrimp culture environment nor is it inherently present in shrimp grow-out ponds. The occurrence of Salmonella bacteria in shrimp from aquaculture operations is related to the concentration of fecal bacteria in the source and grow-out pond water. PMID- 16355821 TI - Campylobacter in food animals and humans in northern Thailand. AB - Cross-sectional, longitudinal, and case-control studies were conducted to describe the epidemiology of Campylobacter in chickens, swine, dairy cows, farm workers, nonfarm residents, and children with diarrhea. Samples were collected in Chiang Mai and Lamphung provinces of northern Thailand from 2000 through 2003. A total of 2,360 samples were processed. Results from the cross-sectional study indicated that the prevalences of Campylobacter in chickens at the farm, slaughterhouse, and market were 64, 38, and 47%, respectively. In swine, the prevalences at the farm, slaughterhouse, and market were 73, 69, and 23%, respectively. Campylobacter prevalence was 14% in dairy cows and 5% in raw milk. The prevalence of Campylobacter on farms was lower in environmental samples than in samples collected from live animals. No Campylobacter isolates were obtained from healthy nonfarm residents, but isolates were obtained from 5 and 18% of farm workers and children with diarrhea, respectively. The prevalence of Campylobacter in pigs in the longitudinal study was 61% at the farm, 46% at the slaughterhouse, and 33% at the market. The majority of Campylobacter isolates from chickens (52%), swine (98%), and farm workers (66%) were Campylobacter coli, whereas the majority of isolates from dairy cows (63%) and children with diarrhea (62%) were Campylobacter jejuni. Most Campylobacter isolates from diarrheal children had single-strand conformation polymorphism profiles similar to those of isolates from chickens. None of the risk factors for infection in children with diarrhea were significantly associated with the isolation of Campylobacter. PMID- 16355823 TI - Virulence and stress susceptibility of clinical and environmental strains of Vibrio vulnificus isolated from samples from Taiwan and the United States. AB - Vibrio vulnificus is an estuarine bacterium that causes severe wound infection and septicemia with high mortality. It also can be transmitted through the consumption of raw contaminated seafood and is an important foodborne pathogen. A total of 40 environmental and clinical V. vulnificus strains isolated from the United States and Taiwan were analyzed for virulence in animals, the presence of virulence-associated factors, and susceptibility to environmental stresses. Virulence in mice was exhibited by 85% of the environmental strains and 95% of the clinical strains. Strains from environmental or clinical sources were similar in virulence-associated phenotypes (protease activity, utilization of transferrin bound iron, hemolysis, and inactivation in serum) and susceptibility to various stresses (4 and 52 degrees C, 0.1 and 10% NaCl, and pH 3.2), except freeze-thaw treatment. The clinical strains killed experimental animals after a shorter incubation time than did the environmental strains. Most of the 15 virulence associated genes examined were present in most of the strains, regardless of their sources or virulence, with the exception of vvh, flgF, and purH. vvh was significantly more common in clinical strains than in environmental strains, and vvh, flgF, and purH were more common in virulent strains than in nonvirulent strains. These data may be helpful in devising strategies to manage or reduce the presence of V. vulnificus in foods. PMID- 16355824 TI - Survival and growth of Enterobacter sakazakii on fresh-cut fruits and vegetables and in unpasteurized juices as affected by storage temperature. AB - Enterobacter sakazakii is an emerging foodborne pathogen that has caused illnesses and deaths in infants and elderly immunocompromised adults. Outbreaks of E. sakazakii infection have been associated with infant formulas, but the documented presence of this pathogen in a wide variety of ready-to-eat foods, including lettuce and other raw vegetables, makes it important to learn more about its behavior in these environments. We investigated the survival and growth characteristics of E. sakazakii on fresh-cut apple, cantaloupe, strawberry, watermelon, cabbage, carrot, cucumber, lettuce, and tomato and in juices prepared from these fruits and vegetables. Produce and juices were inoculated with E. sakazakii at 2 to 3 log CFU/g and 1 to 2 log CFU/ml, respectively, and stored at 4, 12, or 25 degrees C. Populations either did not change or gradually decreased in fresh-cut produce and juices stored at 4 degrees C but grew at 12 degrees C on fresh-cut apple, cantaloupe, watermelon, cucumber, and tomato and in all juices except apple, strawberry, cabbage, and tomato juices. All fresh-cut fruits and vegetables except strawberry supported growth of E. sakazakii at 25 degrees C. Growth occurred in all juices except apple, strawberry, and cabbage juices, followed by decreases in population to < 1 CFU/ml after 48 to 72 h, which coincided with decreases in pH and an increase in the population of lactic acid bacteria. Increases in total counts occurred in all juices except strawberry juice stored at 25 degrees C and apple and strawberry juices stored at 12 degrees C. Total counts increased in cantaloupe, carrot, cucumber, and lettuce juices stored at 4 degrees C. Populations of molds and yeasts increased in apple and tomato juices stored at 25 degrees C but decreased to <1 CFU/ml in cabbage, lettuce, and cucumber juices. Further characterization of the behavior of E. sakazakii on fresh produce and in unpasteurized juice as affected by commercial packaging and handling practices is needed. PMID- 16355825 TI - Outgrowth of Salmonellae and the physical property of albumen and vitelline membrane as influenced by egg storage conditions. AB - This study was undertaken to determine the influence of storage time and temperature on the volume, weight, and pH of egg albumen, the physical strength of vitelline membrane, and the fate of Salmonella Enteritidis artificially inoculated into egg albumen. A fiber-optic probe was used for inoculation with Salmonella Enteritidis at 10(2), 10(4), or 10(6) cells per egg. Both fresh and inoculated eggs were stored at 4, 10, and 22 degrees C for 6 weeks. Five fresh uninoculated eggs from each storage group were collected each week, and the weight, volume, and pH of the egg albumen were measured. The forces, energies, and degrees of membrane deformation required to rupture the vitelline membranes also were determined from either albumen-free yolks or yolks surrounded by albumen. In separate experiments, five inoculated eggs were evaluated each week for populations of Salmonella Enteritidis. When the eggs were stored at 4 degrees C, the albumen retained significantly more volume and weight and had a relatively lower pH. The vitelline membranes from eggs stored at 4 and 10 degrees C required more force and energy for rupture. Salmonellae flourished at 22 degrees C, even in the albumen with the lowest initial population, 10(2) cells per egg. Storage at 4 and 10 degrees C inhibited the growth of salmonellae in the albumen of eggs with initial populations of 10(2), 10(4), or 10(6) cells per egg. In eggs with initial Salmonella populations of 10(6) cells per egg that were stored at 22 degrees C, the populations of reached as high as 10(10) cells per egg after 4 weeks of storage. Storage at 4 and perhaps 10 degrees C postponed the aging process of chicken eggs, preserved the antimicrobial agents of the albumen, and maintained the integrity of vitelline membrane. Low-temperature storage therefore had a significant impact on the safety and overall quality of the eggs. PMID- 16355826 TI - Growth inhibition of Escherichia coli O157:H7 and Listeria monocytogenes by carvacrol and eugenol encapsulated in surfactant micelles. AB - Growth inhibition of four strains of Escherichia coli O157:H7 (H1730, F4546, 932, and E0019) and Listeria monocytogenes (Scott A, 101, 108, and 310) by essential oil components (carvacrol and eugenol) solubilized in nonionic surfactant micelles (Surfynol 465 and 485W) was investigated. Concentrations of encapsulated essential oil components ranged from 0.02 to 1.25% depending on compound, surfactant type, and surfactant concentration (0.5 to 5%). Eugenol encapsulated in Surfynol 485W micelles was most efficient in inhibiting growth of the pathogens; 1% Surfynol 485W and 0.15% eugenol was sufficient to inhibit growth of all strains of E. coli O157:H7 and three of four strains of L. monocytogenes (Scott A, 310, and 108). The fourth strain, L. monocytogenes 101, was inhibited by 2.5% Surfynol and 0.225% eugenol. One percent Surfynol 485W in combination with 0.025% carvacrol was effective in inhibiting three of four strains of E. coli O157:H7. Strain H1730 was the most resistant strain, requiring 0.3% carvacrol and 5% surfactant for complete inhibition. Growth inhibition of L. monocytogenes by combinations of carvacrol and Surfynol 465 ranged between 0.15 and 0.35% and 1 and 3.75%, respectively. Generally, the antimicrobial activity of Surfynol 465 in combination with eugenol was higher than that for the combination with carvacrol. The potent activity was attributed to increased solubility of essential oil components in the aqueous phase due to the presence of surfactants and improved interactions of antimicrobials with microorganisms. PMID- 16355827 TI - Combined effect of carvacrol and packaging conditions on radiosensitivity of Escherichia coli and Salmonella Typhi in ground beef. AB - This study was undertaken to evaluate the effect of 1.0% carvacrol and 0.1% tetrasodium pyrophosphate on the radiation D10 of Escherichia coli and Salmonella Typhi added to ground beef at a concentration of 10(5) CFU/g. The ground beef was then packaged under four different atmosphere conditions: air (78.1% N2, 20.9% O2, 0.036% CO2), 100% CO2, modified atmosphere (60% O2, 30% CO2, 10% N2) and vacuum. Samples were irradiated at doses of 0.1 to 0.6 kGy for E. coli and 0.5 to 2.0 kGy for Salmonella Typhi. Radiation D10-values of 0.126 and 0.526 kGy were observed for E. coli and Salmonella Typhi, respectively, when meat was packed under air. When meat was packed under modified atmosphere conditions, the radiation D10-values for E. coli and Salmonella Typhi were 0.086 and 0.221 kGy, respectively. The addition of carvacrol and tetrasodium pyrophosphate and the use of modified atmosphere packaging reduced the radiation D10-value from 0.126 to 0.046 kGy for E. coli and from 0.526 to 0.053 kGy for Salmonella Typhi. Under vacuum and 100% CO2 conditions, the radiation D10 reduction was not as great as that for the modified atmosphere regardless of the presence or absence of carvacrol and tetrasodium pyrophosphate. PMID- 16355828 TI - Effect of gamma radiation and oregano essential oil on murein and ATP concentration of Escherichia coli O157:H7. AB - This study was carried out to evaluate the effects of gamma radiation alone or in combination with oregano essential oil on the murein composition of Escherichia coli O157:H7 and on the intracellular and extracellular concentrations of ATP. The bacterial strain was treated with three radiation doses: 0.4 kGy to induce cell damage, 1.1 kGy to obtain a viable but non-culturable state, and 1.3 kGy to cause cell death. Oregano essential oil was used at 0.006 and 0.025% (wt/vol), which is the MIC. All treatments had a significant effect (P < or = 0.05) on the murein composition, although some muropeptides did not seem to be affected by the treatment. Each treatment had a different effect on the relative percentage and number of muropeptides. There was a significant correlation (P < or = 0.05) between the decrease in intracellular ATP and the increase in extracellular ATP following treatment of the cells with oregano oil. The reduction of intracellular ATP was even more important when oregano oil was combined with irradiation, but irradiation alone at a high dose (< or = 1.1 kGy) significantly decreased (P < or = 0.05) the internal ATP without affecting the external ATP. Transmission electron microscopic examination revealed that oregano oil and irradiation have an effect on cell wall structure. PMID- 16355829 TI - Identification of Escherichia coli O157:H7 meat processing indicators for fresh meat through comparison of the effects of selected antimicrobial interventions. AB - Fresh meat products can become contaminated with the pathogen Escherichia coli O157:H7 during the slaughter process; therefore, an E. coli O157:H7 indicator to verify the effectiveness of process controls in slaughter establishments would be extremely useful. The hides of 20 beef cattle were sampled, and 113 bacterial isolates were obtained. Thirteen of these isolates representing four genera, Escherichia, Enterobacter, Providencia, and Serratia, were selected based on growth and biochemical characteristics similar to those of five clinical strains of E. coli O157:H7. The temperature sensitivity was determined for the individual isolates and the five E. coli O157:H7 strains at 55 and 65 degrees C. D65-values for all 13 isolates were not significantly different from D65-values of the E. coli O157:H7 strains. E. coli isolates were the only isolates whose D55-values were not significantly different from those of the E. coli O157:H7 strains. E. coli isolates P3 and P68 were more resistant to the effects of 55 degrees C than were the other E. coli isolates but were not significantly different from E. coli O157:H7 WS 3331 (P > 0.05). The remaining E. coli isolates (P1, P8, and P14) were not significantly different from E. coli O157:H7 strains ATCC 35150, ATCC 43894, ATCC 43895, and WS 3062 (P > 0.05). Prerigor lean and adipose beef carcass tissue was artificially contaminated with stationary-phase cultures of the five E. coli beef cattle isolates or a cocktail of five E. coli O157:H7 strains in a fecal inoculum. Each tissue sample was processed with the following microbial interventions: 90 degrees C water; 90 degrees C water followed by 55 degrees C 2% lactic acid; 90 degrees C water followed by 20 degrees C 2% lactic acid; 20 degrees C water followed by 20 degrees C 2% lactic acid; 20 degrees C water followed by 20 degrees C 20 ppm chlorine; and 20 degrees C water followed by 20 degrees C 10% trisodium phosphate. The appropriateness of the E. coli isolates as potential E. coli O157:H7 indicators was dependent upon the microbial intervention utilized. For all microbial intervention methods applied irrespective of tissue type, the mean log reductions of at least two E. coli isolates were not significantly different from the mean log reduction of the E. coli O157:H7 cocktail (P > 0.05). Because of the frequent employment of multiple microbial interventions in the cattle industry, no single isolate can realistically represent the effectiveness of all microbial interventions for reduction of E. coil O157:H7. Thus, the use of a combination of E. coli isolates may be required to accurately predict the effectiveness of microbial intervention methods on the reduction of E. coli O157:H7 in beef carcass tissue. PMID- 16355830 TI - Influence of blanching treatments on Salmonella during home-type dehydration and storage of potato slices. AB - Recommended drying treatments may not enhance destruction of pathogens that could be present on home-dried foods. In this study, the effects of traditional and modified treatments on Salmonella were evaluated during preparation, home-type dehydration (60 degrees C for 6 h), and storage of potato slices. Potato slices inoculated with five strains of Salmonella (8.4 log CFU/ g) were left untreated or were treated by steam blanching (88 degrees C for 10 min), water blanching (88 degrees C for 4 min), 0.105% citric acid blanching (88 degrees C for 4 min), or 0.210% citric acid blanching (88 degrees C for 4 min). Slices were then dried (6 h for 60 degrees C) and aerobically stored for up to 30 days at 25 +/- 3 degrees C. Cells were enumerated on tryptic soy agar with 0.1% pyruvate (TSAP) and on xylose lysine deoxycholate agar. Salmonella populations were reduced by 4.5 to 4.8 CFU/g and by >5.4 log CFU/g immediately following steam and water blanching, respectively. Populations were below the detection limit (0.80 log CFU/g) immediately following acid blanching, except for samples blanched in 0.105% citric acid and recovered on TSAP. After dehydration (6 h for 60 degrees C), Salmonella reductions on blanched potato slices (5.3 to 5.6 log CFU/g) were significantly greater (P < 0.05) than those on untreated samples (1.9 to 2.7 log CFU/g). Populations on all samples continued to decrease throughout 30 days of storage but still were 3.1 to 3.9 log CFU/g on untreated samples. In comparison, bacterial populations on blanched samples were undetectable by direct plating following 30 days of storage (regardless of blanching method). Blanching treatments used in this study improved the effectiveness of drying for inactivating Salmonella inoculated onto potato slices and, therefore, may enhance the safety of the product. PMID- 16355831 TI - Predictive model for Clostridium perfringens growth in roast beef during cooling and inhibition of spore germination and outgrowth by organic acid salts. AB - Spores of foodborne pathogens can survive traditional thermal processing schedules used in the manufacturing of processed meat products. Heat-activated spores can germinate and grow to hazardous levels when these products are improperly chilled. Germination and outgrowth of Clostridium perfringens spores in roast beef during chilling was studied following simulated cooling schedules normally used in the processed-meat industry. Inhibitory effects of organic acid salts on germination and outgrowth of C. perfringens spores during chilling and the survival of vegetative cells and spores under abusive refrigerated storage was also evaluated. Beef top rounds were formulated to contain a marinade (finished product concentrations: 1% salt, 0.2% potassium tetrapyrophosphate, and 0.2% starch) and then ground and mixed with antimicrobials (sodium lactate and sodium lactate plus 2.5% sodium diacetate and buffered sodium citrate and buffered sodium citrate plus 1.3% sodium diacetate). The ground product was inoculated with a three-strain cocktail of C. perfringens spores (NCTC 8238, NCTC 8239, and ATCC 10388), mixed, vacuum packaged, heat shocked for 20 min at 75 degrees C, and chilled exponentially from 54.5 to 7.2 degrees C in 9, 12, 15, 18, or 21 h. C. perfringens populations (total and spore) were enumerated after heat shock, during chilling, and during storage for up to 60 days at 10 degrees C using tryptose-sulfite-cycloserine agar. C. perfringens spores were able to germinate and grow in roast beef (control, without any antimicrobials) from an initial population of ca. 3.1 log CFU/g by 2.00, 3.44, 4.04, 4.86, and 5.72 log CFU/g after 9, 12, 15, 18, and 21 h of exponential chilling. A predictive model was developed to describe sigmoidal C. perfringens growth curves during cooling of roast beef from 54.5 to 7.2 degrees C within 9, 12, 15, 18, and 21 h. Addition of antimicrobials prevented germination and outgrowth of C. perfringens regardless of the chill times. C. perfringens spores could be recovered from samples containing organic acid salts that were stored up to 60 days at 10 degrees C. Extension of chilling time to > or =9 h resulted in >1 log CFU/g growth of C. perfringens under anaerobic conditions in roast beef. Organic acid salts inhibited outgrowth of C. perfringens spores during chilling of roast beef when extended chill rates were followed. Although C. perfringens spore germination is inhibited by the antimicrobials, this inhibition may represent a hazard when such products are incorporated into new products, such as soups and chili, that do not contain these antimicrobials, thus allowing spore germination and outgrowth under conditions of temperature abuse. PMID- 16355832 TI - Development and validation of primary, secondary, and tertiary models for growth of Salmonella Typhimurium on sterile chicken. AB - Models are used in the food industry to predict pathogen growth and to help assess food safety. However, criteria are needed to determine whether models provide acceptable predictions. In the current study, primary, secondary, and tertiary models for growth of Salmonella Typhimurium (10(4.8) CFU/g) on sterile chicken were developed and validated. Kinetic data obtained at 10 to 40 degrees C were fit to a primary model to determine initial density (N0), lag time (lambda), maximum specific growth rate (micromax), and maximum population density (Nmax). Secondary models for N0, lambda, micromax, and Nmax as a function of temperature were developed and combined with the primary model to create a tertiary model that predicted pathogen density (N) at times and temperatures used and not used in model development. Performance of models was evaluated using the acceptable prediction zone method in which experimental error associated with growth parameter determinations was used to set criteria for acceptable model performance. Models were evaluated against dependent and independent (validation) data. Models with 70% prediction or relative errors (RE) in an acceptable prediction zone from -0.3 to 0.15 for micromax, -0.6 to 0.3 for lambda, and -0.8 to 0.4 for N, N0, and Nmax were classified as acceptable. All secondary models had acceptable goodness of fit and were validated against independent (interpolation) data. Percent RE in the acceptable prediction zone for the tertiary model was 90.7 for dependent data and 97.5 for independent (interpolation) data. Although the tertiary model was validated for interpolation, an unacceptable %RE of 2.5 was obtained for independent (extrapolation) data obtained with a lower N0 (10(0.8) CFU/g). The tertiary model provided overly fail-dangerous predictions of N from a lower N0. Because Salmonella concentrations on chicken are closer to 10(0.8) than 10(4.8) CFU/g, the tertiary model should not be used to help assess chicken safety. PMID- 16355833 TI - Biofilm formation and sporulation by Bacillus cereus on a stainless steel surface and subsequent resistance of vegetative cells and spores to chlorine, chlorine dioxide, and a peroxyacetic acid-based sanitizer. AB - Biofilm formation by Bacillus cereus 038-2 on stainless steel coupons, sporulation in the biofilm as affected by nutrient availability, temperature, and relative humidity, and the resistance of vegetative cells and spores in biofilm to sanitizers were investigated. Total counts in biofilm formed on coupons immersed in tryptic soy broth (TSB) at 12 and 22 degrees C consisted of 99.94% of vegetative cells and 0.06% of spores. Coupons on which biofilm had formed were immersed in TSB or exposed to air with 100, 97, 93, or 85% relative humidity. Biofilm on coupons immersed in TSB at 12 degrees C for an additional 6 days or 22 degrees C for an additional 4 days contained 0.30 and 0.02% of spores, respectively, whereas biofilm exposed to air with 100 or 97% relative humidity at 22 degrees C for 4 days contained 10 and 2.5% of spores, respectively. Sporulation did not occur in biofilm exposed to 93 or 85% relative humidity at 22 degrees C. Treatment of biofilm on coupons that had been immersed in TSB at 22 degrees C with chlorine (50 microg/ml), chlorine dioxide (50 microg/ml), and a peroxyacetic acid-based sanitizer (Tsunami 200, 40 microg/ml) for 5 min reduced total cell counts (vegetative cells plus spores) by 4.7, 3.0, and 3.8 log CFU per coupon, respectively; total cell counts in biofilm exposed to air with 100% relative humidity were reduced by 1.5, 2.4, and 1.1 log CFU per coupon, respectively, reflecting the presence of lower numbers of vegetative cells. Spores that survived treatment with chlorine dioxide had reduced resistance to heat. It is concluded that exposure of biofilm formed by B. cereus exposed to air at high relative humidity (> or =97%) promotes the production of spores. Spores and, to a lesser extent, vegetative cells embedded in biofilm are protected against inactivation by sanitizers. Results provide new insights to developing strategies to achieve more effective sanitation programs to minimize risks associated with B. cereus in biofilm formed on food contact surfaces and on foods. PMID- 16355834 TI - Economic cost of illness due to Escherichia coli O157 infections in the United States. AB - The Centers for Disease Control and Prevention (CDC) has estimated that Shiga toxin-producing Escherichia coli O157 (0157 STEC) infections cause 73,000 illnesses annually in the United States, resulting in more than 2,000 hospitalizations and 60 deaths. In this study, the economic cost of illness due to O157 STEC infections transmitted by food or other means was estimated based on the CDC estimate of annual cases and newly available data from the Foodborne Diseases Active Surveillance Network (FoodNet) of the CDC Emerging Infections Program. The annual cost of illness due to O157 STEC was $405 million (in 2003 dollars), including $370 million for premature deaths, $30 million for medical care, and $5 million in lost productivity. The average cost per case varied greatly by severity of illness, ranging from $26 for an individual who did not obtain medical care to $6.2 million for a patient who died from hemolytic uremic syndrome. The high cost of illness due to O157 STEC infections suggests that additional efforts to control this pathogen might be warranted. PMID- 16355835 TI - Relationship between consumer food safety knowledge and reported behavior among students from health sciences in one region of Spain. AB - A survey was conducted to investigate the relationship between knowledge about food safety and actual food handling practices among Spanish university students (mainly from the health sciences disciplines) who usually prepare meals at home. Based on level of education in food safety topics, students were divided in three groups: high, which included students from Food Science and Nutrition; medium, which included students from other health sciences; and low, which included students from non-health-related studies. More than two thirds of the 562 people selected had an accurate knowledge of the eight foodborne pathogens included in the survey, but only 5.2% were able to identify Staphylococcus aureus as a foodborne pathogen. Significant differences in responses were found depending on educational level concerning the food safety topic. For food handling, up to 60% of the responses reflected accurate knowledge of proper storage of prepared meals and washing of hands and materials to avoid cross-contamination. However, with the exception of questions related to storage temperature, there was considerable difference between knowledge and reported behavior. Although 98.6% of the participants recognized the importance of hand washing before and during food preparation, only one quarter (24.4%) affirmed that they washed their hands with soap and water. On questions concerning food practices, more accurate answers were given by the older students. Women answered questions regarding cross contamination more accurately, whereas men were more accurate in response to questions concerning temperature and food preservation. In general, students with more knowledge of food hygiene had better reported practices, but even these students reported some high-risk behaviors. These results confirm the need to improve educational programs, ensuring that the acquired knowledge actually modifies consumer behavior. PMID- 16355836 TI - Evaluation of detection methods for screening meat and poultry products for the presence of foodborne pathogens. AB - Rapid and molecular technologies such as enzyme-linked immunosorbent assay (ELISA), PCR, and lateral flow immunoprecipitation can reduce the time and labor involved in screening food products for the presence of pathogens. These technologies were compared with conventional culture methodology for the detection of Salmonella, Campylobacter, Listeria, and Escherichia coli O157:H7 inoculated in raw and processed meat and poultry products. Recommended protocols were modified so that the same enrichment broths used in the culture methods were also used in the ELISA, PCR, and lateral flow immunoprecipitation assays. The percent agreement between the rapid technologies and culture methods ranged from 80 to 100% depending on the pathogen detected and the method used. ELISA, PCR, and lateral flow immunoprecipitation all performed well, with no statistical difference, compared with the culture method for the detection of E. coli O157:H7. ELISA performed better for the detection of Salmonella, with sensitivity and specificity rates of 100%. PCR performed better for the detection of Campylobacter jejuni, with 100% agreement to the culture method. PCR was highly sensitive for the detection of all the foodborne pathogens tested except Listeria monocytogenes. Although the lateral flow immunoprecipitation tests were statistically different from the culture methods for Salmonella and Listeria because of false-positive results, the tests did not produce any false negatives, indicating that this method would be suitable for screening meat and poultry products for these pathogens. PMID- 16355837 TI - Multicenter validation of a multiplex PCR assay for differentiating the major Listeria monocytogenes serovars 1/2a, 1/2b, 1/2c, and 4b: toward an international standard. AB - The performance of a multiplex PCR assay that separates the four major serovars of the pathogenic Listeria monocytogenes into four distinct PCR groups was evaluated through a multicenter typing study. Identical panels of 90 Listeria isolates were distributed to five participating laboratories that were blind to the nature of the isolates. Isolates were characterized using the previously standardized protocol. Overall concordance was 96.6 to 100%, sufficient for the assay to be used as an alternative to serotyping and confidently applied in laboratories involved in L. monocytogenes typing. PMID- 16355838 TI - Evaluation of a rapid PCR-based method for the detection of animal material. AB - A rapid PCR-based analytical method for detection of animal-derived materials in complete feed was developed. Using a commercially available DNA forensic kit for the extraction of DNA from animal feed, a sensitive method was developed that was capable of detecting as little as 0.03% bovine meat and bone meal in complete feed in under 8 h of total assay time. The reduction in assay time was accomplished by reducing the DNA extraction time to 2 h and using the simpler cleanup procedure of the kit. Assay sensitivity can be increased to 0.006% by increasing the DNA extraction time to an overnight incubation of approximately 16 h. Examination of dairy feed samples containing either bovine meat and bone meal, porcine meat and bone meal, or lamb meal at a level of 0.1% (wt/wt basis) suggested that this method may be suitable for regulatory uses. The adoption of this commercially available kit for use with animal feeds yields an assay that is quicker and simpler to perform than a previously validated assay for the detection of animal proteins in animal feed. PMID- 16355839 TI - Evaluation of two commercial lateral-flow test kits for detection of animal proteins in animal feed. AB - Performance characteristics were evaluated for two lateral-flow test kits, Reveal for Ruminant in Feed (Neogen Corporation) and FeedChek (Strategic Diagnostics Inc.), designed to detect ruminant or terrestrial animal proteins in feeds. The stringent acceptance criteria used were developed by the Center for Veterinary Medicine Office of Research to identify test kits with comparable selectivity and sensitivity to microscopy and PCR assay, the analytical methods used by the U.S. Food and Drug Administration (FDA). Guidelines were developed for evaluating the selectivity, sensitivity, ruggedness, and specificity of these kits. These guidelines further stated that ruggedness and specificity testing would be performed only after a test passed both the selectivity and sensitivity assessments. Acceptance criteria for determining success were developed using a statistical approach requiring 90% probability of achieving the correct response, within a 95% confidence interval. A minimum detection level of 0.1% bovine meat and bone meal, consistent with the sensitivity of the methods used by the FDA, was required. Selectivity was assessed by testing 60 dairy feed samples that contained no added animal proteins; sensitivity was determined by evaluating 60 samples (per level of fortification) of the same feed that contained 0.025, 0.05, 0.1, 0.25, 0.5, 1, or 2% bovine meat and bone meal. The Reveal test passed the selectivity assessment but failed the sensitivity assessment, detecting only samples fortified at the 2% level and then only 17 to 33% of those samples, when read according to the label directions. The FeedChek test passed the sensitivity assessment but failed the selectivity assessment, with rates for false-positive results ranging from 34 to 38%, depending on the user. The sensitivity of the Reveal test was affected by the concentration of trace minerals present in the feed; concentrations toward the high end of the normal range prevented the detection of true positive feed samples containing bovine meat and bone meal. Better sensitivity assessments were obtained when lamb meal was used either alone or in combination with bovine meat and bone meal. The FeedChek test was not affected by the concentration of trace minerals or by the type of animal meal used. These results indicate that neither of the two tests is adequate for routine regulatory use. PMID- 16355840 TI - A washed-curd goat's cheese as a vehicle for delivery of a potential probiotic bacterium: Lactobacillus delbrueckii subsp. lactis UO 004. AB - This study characterizes the probiotic properties of Lactobacillus delbrueckii subsp. lactis UO 004 and examines its suitability for making cheese. This strain was isolated from infant feces and shows interesting features, such as acid and bile tolerance, adherence to intestinal epithelial cells, and inhibition of the growth of certain enteropathogens, that support its potential use as a probiotic strain. In this regard, the suitability of a washed-curd cheese (Vidiago type) made with goat's milk as a delivery system for this probiotic strain was assessed. Lactobacillus delbrueckii subsp. lactis UO 004 was incorporated into a starter culture (IPLA 001). Changes in the overall composition of control and experimental cheeses were determined during ripening through bacteriological, chemical, high-performance liquid chromatography, and gas chromatography analyses. Slight changes in the gross composition and appreciable differences in the flavor compounds profile were observed between control and experimental cheeses. This strain was capable of surviving at high cell numbers (10(8) to 10(9) CFU/g) in cheeses after 28 days of ripening without adversely affecting sensory criteria or appearance of the cheese, thus satisfying the criteria for a probiotic food product. PMID- 16355841 TI - Adhesion of selected Bifidobacterium strains to human intestinal mucus and the role of adhesion in enteropathogen exclusion. AB - The ability of potential probiotic strains to adhere to the intestinal mucosa and exclude and displace pathogens is of utmost importance for therapeutic manipulation of the enteric microbiota. The ability of seven selected human bifidobacterial strains and five human enteropathogenic strains to adhere to human intestinal mucus was analyzed and compared with that of four strains isolated from chicken intestines. The adhesion of the bifidobacterial strains ranged from 3 to 16% depending on the strain. Bifidobacterium strains of animal origin adhered significantly better than did strains of human origin. Of the pathogenic bacteria, Escherichia coli NCTC 8603 had the highest adhesion value (20%), Salmonella Typhimurium ATCC 29631, Enterobacter sakazakii ATCC 29544, and Clostridium difficile ATCC 9689 had adhesion values ranging from 10 to 15%, and Listeria monocytogenes ATCC 15313 had the lowest adhesive value (3%). The ability of these bifidobacteria to inhibit pathogen adhesion and to displace pathogens previously adhering to mucus was also tested. The inhibition of pathogens adhesion by these bifidobacterial strains was variable and clearly strain dependent. In general, bifidobacterial strains of animal origin were better able to inhibit and displace pathogens than were human strains. Preliminary characterization of bacterial adhesion was accomplished using different pretreatments to explore adhesion mechanisms. The results indicate that different molecules are implicated in the adhesion of bifidobacteria to the human intestinal mucus, constituting a multifactorial process. PMID- 16355842 TI - Predicting toxicity equivalents in fish tissue. AB - Toxicity equivalents is a measure of "dioxin-like" toxicity contributed by polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (dioxins), and polychlorinated dibenzofurans (furans). Calculation of toxicity equivalents require the use of analytical procedures that are expensive and slow, making them impractical for routine analysis. U.S. Environmental Protection Agency National Fish Tissue Study (2003) data were used to determine the most significant predictors of toxicity equivalents with multiple regression analysis. The strongest predictive model (P < 0.0001, R2 = 0.97) included five compounds (PCB 118; PCB-126; 2,3,7,8-TCDD; 1,2,3,6,7,8-HxCDD; 2,3,4,7,8-PeCDF). However, the required lower limit of detection for an analytical method measuring these congeners is 0.1 ppt and would not provide much benefit over the current analytical method. An alternative model (P < 0.0001, R2 = 0.68) that included three PCBs (PCB-138, PCB-153, PCB-118) would require a limit of detection of 1,000 ppt and be more practical. This research demonstrates that the measurement of selected compounds can be used to estimate toxicity equivalents and consequently serve as the impetus for the development of lower cost, rapid analytical methods for analysis of fish. PMID- 16355843 TI - Shelf life of fresh sausages stored under modified atmospheres. AB - The aim of this study was to investigate differences in modified atmosphere packaging (MAP) for the improvement of the shelf life of fresh meat products. Three different conditions for preserving fresh sausages were tested: MAP1 (20% CO2, 70% O2, and 10% N2), MAP2 (40% CO2 and 60% O2), and MAP3 (40% CO2, 30% O2, and 30% N2). Samples from the MAP2 group had fewer spoilage bacteria, stable red color (no change of a* value), and good physical attributes (high water-holding capacity, little loss from cooking, and low shear force needed for cutting) compared with samples from other treatment groups. Thus, high concentrations of CO2 (40%) and O2 (60%) resulted in a longer shelf life for fresh sausages. PMID- 16355844 TI - Prevalence of class 1 integrons and antimicrobial resistance gene cassettes among enteric bacteria found in multisite group-level cohorts of humans and swine. AB - The prevalence of antimicrobial resistance genotypic characteristics (class 1 integrons and antimicrobial resistance gene cassettes) among commensal Escherichia coli isolated from humans and swine in a semiclosed, integrated farrow-to-fork population was evaluated in a cross-sectional study. The objective of this study was to establish baseline antimicrobial resistance patterns of enteric bacteria from animals and humans within the study population; specifically, genotypic traits both unique and common to commensal E. coli derived from the different sources were evaluated. There were significant differences between host species; swine isolates were more likely to harbor integrons (odds ratio = 2.33, P = 0.0487). No significant differences were found for facility location, facility type, human housing cohort, or time of day (P > 0.05). There were significant differences (P = 0.006) among swine production groups (fecal samples from boars, dry sows, finishers, growers, intake boars, lactating sows, the lagoon, nursery piglets, influent, and piglets); the grower group was less likely than the nursery group to harbor a class 1 integron (nursery as referent: odds ratio = 0.22, P = 0.04). Among all isolates with an integron present, human isolates were more likely to harbor an antimicrobial resistance gene cassette (odds ratio = 6.36, P = 0.003). When isolates that possessed gene cassettes coding for resistance to specific antimicrobials were compared, no significant differences between host species (P > 0.05) were observed. PMID- 16355845 TI - Movement and persistence of Salmonella in broiler chickens following oral or intracloacal inoculation. AB - The dissemination of Salmonella into various lymphoid-like organs in young broiler chicks after oral and intracloacal inoculation was studied. A three strain cocktail of Salmonella Typhimurium, Salmonella Montevideo, and Salmonella Enteritidis was administered either orally or intracloacally to day-old chicks. After 1 h, 1 day, or 1 week, the ceca, thymus, liver and gallbladder, spleen, and bursa were sampled for the presence of Salmonella. There was a marked difference in the recovery of Salmonella 1 h postinoculation. Only 6 of 50 samples from orally inoculated chicks were positive compared with 33 of 50 samples from cloacally inoculated samples. In comparison, 24 h and 1 week after inoculation, there was no difference in the number of positive samples between oral or cloacal inoculation. The rapidity of the translocation of the Salmonella from the cloacal inoculum compared to the oral inoculum is likely due to the transient time required for Salmonella to move through the alimentary tract. The method of inoculation did not affect the distribution of serogroups. Of the three serotypes in the composite inoculum, the Salmonella Enteritidis (group D) was recovered only twice in replication 1 and not at all in replication 2. Both the Salmonella Typhimurium (serogroup B) and the Salmonella Montevideo (serogroup C1) were recovered extensively throughout the study. PMID- 16355846 TI - Salmonella enterica serotypes isolated from imported frozen chicken meat in the Canary islands. AB - To determine the prevalence of Salmonella enterica serotypes in imported frozen chicken meat, 406 samples (whole chicken, legs, and breast meat) were analyzed for Salmonella according to ISO6579 rules, serotypes were assigned, and phage typing was conducted for Salmonella serotypes Enteritidis, Typhimurium, and Heidelberg. The overall frequency of Salmonella isolation was 16.5%. By country of origin, the highest percentage of cases was found among the samples from France followed by samples from Brazil. The differences between legs and breast meat were significant. The most frequently isolated serotype of Salmonella was Enteritidis, followed by Salmonella Heidelberg, Salmonella Typhimurium, and Salmonella Virchow. By country of origin, we identified a large percentage of serotype Salmonella Enteritidis in the samples imported from Brazil. There was a greater diversity of serotypes isolated from the French samples, and Salmonella Enteritidis was not the dominant strain. In the samples from the United States, the only serotype isolated was Salmonella Kentucky, although a smaller number of samples was analyzed. The Salmonella Enteritidis phage type that prevailed in both France and Brazil was 4. Phage types 204c and 204 were identified for Salmonella Typhimurium, and phage types 8, 31, and 37 were identified for Salmonella Virchow. PMID- 16355847 TI - Subtyping Listeria monocytogenes from bulk tank milk using automated repetitive element-based PCR. AB - Sixty-one Listeria monocytogenes strains from raw milk were analyzed with an automated repetitive element-based PCR (rep-PCR) system to examine the utility of this system for serotype grouping and to determine whether specific regional relationships could be identified. Results of the similarity analysis revealed two primary clusters of L. monocytogenes isolates. Cluster 2 exclusively contained serogroup 1/2a isolates; however, two 1/2a isolates were also found in cluster 1. Isolates of serogroups 1/2b, 4b, 3b, and 4c were also in cluster 1. Clusters 1 and 2 were separated at a relative similarity of 86%. Listeria species other than L. monocytogenes (L. ivanovii, L. seeligeri, L. welshimeri, L. grayi, and L. innocua) had similarity scores of less than 80% in pairwise comparisons with the L. monocytogenes isolates. Thus, this method may be useful for species identification once an isolate is characterized as Listeria. When rep-PCR fingerprints of the L. monocytogenes 1/2a isolates were compared, there was no apparent regional grouping. However, discrimination between isolates suggests that the rep-PCR assay might be useful for tracking L. monocytogenes 1/2a and for tracking isolates across regions or within smaller ecological niches. The automated rep-PCR method could not discriminate between serotypes 1/2b and 4b but may be useful for discriminating between 1/2a and other serotypes and for tracking isolates within serotype 1/2a. PMID- 16355848 TI - Antifungal activity of Mexican oregano (Lippia berlandieri Shauer). AB - Antifungal and sensorial properties of spices have been recognized for years. The antifungal compounds are products of the plant's secondary metabolism, and the action of those compounds could be used to inhibit the growth of spoilage and pathogenic microorganisms in food. Mexican oregano (Lippia berlandieri) grows wildly in the desert zone of Mexico and is usually added to regional foods. The goal of this study was to evaluate the antifungal activity of Mexican oregano versus food-contaminant fungi. Fungi were isolated from spoiled fruit and vegetables and identified according to morphological characteristics. The antifungal activity of oregano was evaluated by radial growth measurement on potato dextrose agar added with dried oregano (0.25 to 4.0%). The essential oil antifungal activity of oregano was also evaluated by the diffusion well test. Twenty-one fungal strains were isolated, which included Penicillium, Geotrichum, Aspergillus, and Bipolaris. In seven of the 21 strains, no inhibitory effect was observed at either concentration of oregano. An increase in growth at the lower or higher concentrations of oregano, when compared to the control, was observed in two fungal strains; in 12 strains, a strong inhibitory effect of oregano was evident. The oregano essential oil was inhibitory to all fungal strains, but there were differences in the extent of the effect. Although the antifungal effect of oregano is strongly established, there was a differential effect with the fungal strains studied. Besides pathogenic fungi and bacteria, microbial spoilage flora should be considered when the addition of spices for food preservation is proposed. PMID- 16355849 TI - Microbiological sampling of poultry carcass portions by excision, rinsing, or swabbing. AB - Groups of 25 skin-on thighs or skin-on, skinned, or tumbled breast portions of broiler chicken carcasses were sampled by excision of skin or muscle tissue, rinsing, or swabbing. Counts of total aerobic bacteria, coliforms, and Escherichia coli were recorded for each sample. For all types of carcass portions, the mean log counts and the total log counts obtained for each group of bacteria by excision or rinsing mostly differed by <0.5 log unit. However, the counts obtained by swabbing were generally >0.5 log unit lower than the smaller of the values obtained by the other two sampling methods. PMID- 16355851 TI - Thyroid intrafollicular neoplasia (TIN) as a precursor of papillary microcarcinoma. AB - BACKGROUND: The aim of this study was to identify a spectrum of morphological changes from benign reactive follicles to suspected thyroid intrafollicular neoplasia (TIN) and papillary microcarcinoma observed in different pathological processes of the thyroid gland. CASE SERIES: In a retrospective study on a total of 105 cases, occult or incidental thyroid papillary microcarcinoma was found in association with multinodular goiter (52%), chronic thyroiditis (19%), follicular adenoma (19%), parathyroid neoplasia (5%), and no evident thyroid pathology (5%). RESULTS: Atypical follicles are described in various pathological conditions correlating with lymphoid inflammatory infiltration, stromal sclerosis, and hyperplastic or neoplastic follicular nodules. The histological pattern of papillary carcinoma is compared with morphological changes ranging from atrophic and/or reactive thyroid follicles, potential intrafollicular thyroid neoplasia (TIN) and papillary microcarcinoma. CONCLUSIONS: The atypical features of follicular epithelial areas make up a wide spectrum that starts with the dispersion of nuclear chromatin. Low- and high-grade TIN may represent a cytological marker of dysplastic lesions and precursors of thyroid papillary carcinoma when it is still in an early intrafollicular or pre-invasive stage in various pathological processes of the thyroid. PMID- 16355850 TI - [Bronchial carcinoid: a review of the recent literature]. AB - OBJECTIVE: To evaluate methods of diagnosis and treatment, and the long term survival of patients treated for bronchial carcinoid tumor by the review of recent literature. METHODS: The Authors conducted a retrospective study on internet-based-evidence of patients treated for bronchial carcinoid tumor since 1993 to 2004. Symptoms, diagnosis, operative approach and survival were assessed. CONCLUSIONS: On the basis of the review of the international literature, the Authors affirm that the 5 and 10-year probability of survival are closely linked to the histological type of carcinoid, to the presence of nodal and distant metastasis. Owing to the potential malignancy of these tumours, preference should be given to radical exeresis. PMID- 16355852 TI - [Major thoracic trauma--sternal trauma]. AB - From 1999 to 2003, 101 patients (M/F = 83/18, mean age = 46 +/- 18 y) with thoracic trauma have been admitted to the Center for Thoracic Surgery in Varese. Over 50% of pts. with major thoracic trauma were treated by chest tube drainage; however, surgery was necessary just in 16/101 pts.; 29 pts. were admitted to ICU; deaths were 3/101. The Authors recorded 23/101 sternal fractures. Routine blood test, chest radiography and EKG were performed in all patients; in case of abnormal EKG, echocardiography was performed; in case of widened mediastinum, chest CT was mandatory. 13/23 pts. showed associated injuries, 12 pts. a non aligned sternal fracture, 7 pts. mediastinal hematoma. Simple observation in hospital was indicated in 15/23 pts. with sternal fractures; chest tubes were positioned in 3 pts. (hemopneumothorax) and one patient was surgically treated because of painful sternal pseudoarthrosis (2 months after trauma). Admission in ICU was necessary in 3 pts. with non-aligned sternal fracture, mediastinal hematoma and associated injuries. No patient with sternal fracture died. PMID- 16355853 TI - [Videolaparoscopic cholecystectomy: personal experience]. AB - AIM: Laparoscopic cholecystecomy (LC) is now the gold standard for the treatment of gallstones. In this report were analyzed 153 LC performed by a single surgical team and, according to results, elaborated any technical recommendation. MATERIALS AND METHODS: 153 LC has been performed in Section of General and Thoracic Surgery of University of Palermo, Sicily, by a single surgical team, since 2000. Indications for LC were all forms of calculous cholecystitis (biliary colic in 73.2%, acute cholecystitis in 4.5%, gallbladder polyps in 5.8%) or colecystocoledochal lithiasis (9.8%). The mean age of patients was 49.46 years (range 18-78) and 62.7% were female. The patients were studied in our Section with hematochemical routine, plane chest roentgram, ECG, abdominal ultrasound (the day before the operation) and ASA classification (ASA I: 9.8%, ASA II: 67.3%, ASA III: 22.9%). RESULTS: All cases (153) were uneventful in terms of either serious intraoperative complications or necessity in relaparoscopy. In operatory room we adopted the French position, with mean operatory time of 65.03 minutes (range 30-180 minutes) and we also used the French technique of exposure of the cystic pedicle. The grade of difficulty of LC was analysed according 4 variables (approach to peritoneum, approach to gallbladder, pedicle dissection, cholecystectomy). There were 10 cases (6.5%) of convertions in total. Subhepatic space drainage was performed in 60.1% of cases. No postoperative biloma or subhepatic infiltration were observed. No port site infections were diagnosed, no port site hernias observed and no mortality observed. The mean postoperative bedstay was 2.21 days. CONCLUSIONS: LC is the treatment of choice for symptomatic gallstone disease. When performed by experienced surgeons, it is safe and effective. PMID- 16355854 TI - Minimally invasive approach of the most common complications after laparoscopic cholecystectomy. AB - AIM OF THE STUDY: LC is now considered the gold standard in the treatment of cholelithiasis with a minimal incidence of complications, typically, bile peritoneum and hemoperitoneum. METHODOLOGY: The Authors wished to verify the possibility of treating complications after LC with a minimally invasive approach. In 1100 LCs over 8 years, they treated 9 bile peritoneum and 4 hemoperitoneum. RESULTS: It was possible to treat all 13 patients with a minimally invasive approach with a complete resolution of the complication and with no further occurrences during the follow up. CONCLUSIONS: The Authors believe that it is possible to treat the majority of complications arising after LC with minimally invasive methods. Nevertheless, a very early diagnosis and the close cooperation of an Endoscopist and an Radiologist is necessary. PMID- 16355855 TI - [Role of the severity score and of the multiple organ dysfunctions in the treatment of severe acute pancreatitis and its infective complications]. AB - INTRODUCTION: There is considerable controversy about timing of surgery in acute pancreatitis. Scoring system is widely used in Intensive Care Units (ICU) but above all in order to assess the severity of disease. AIM OF THE STUDY: In this prospective clinical study, modification of clinical evolution and scores (APACHE II, SAPS II and SOFA) are analyzed. Scores were computed, daily or every other day. Two particular phases were observed: the day of ICU admission and the day of surgical treatment. MATERIAL AND METHODS: Twenty-one patients are studied; they were all surgically treated only after identification of positive pancreatic coltures. Open-packing was performed 2-6 weeks after the beginning of acute pancreatitis. In six patients who died, a progressive deterioration was noticed between admission and identification of infected necrosis. All scores were higher in patients who died, in particular a significant difference was found between SOFA score at admission and at treatment (mean +/- SD: 5.0 +/- 3.2 vs 8.5 +/- 3.0, p < 0.05). CONCLUSIONS: In conclusion when clinical conditions are critical a high SOFA score could contribute to indicate surgical treatment even without identification of sepsis. PMID- 16355856 TI - [Diagnostic and therapeutic approach to pancreatic trauma]. AB - INTRODUCTION: The pancreatic trauma is rare, compared with the injuries of the other abdominal organs and occurs in 0.2-6 per cent of the cases of abdominal trauma. AIM OF THE STUDY: The aim of this essay is to demonstrate the Authors' experience in the treatment of five cases of pancreatic injury during the last five years, as well as to retrospect the contemporary bibliography, connected with the diagnostic and curative approach of the pancreatic trauma. The diagnosis of the pancreatic trauma is difficult and many times, late. In their experience of pancreatic trauma, the Authors ascertained the pancreatic injury during the laparotomy which was made in order to treat other abdominal injuries. MATERIAL AND METHODS: The surgical techniques were chosen taking into account the extent of the injury, the detection and the existence of accompanying. Marginal resection of pancreas, splenectomy and drainage were applied to three patients, suture of the pancreas and drainage to one patient, drainage alone and treatment of synchronous rupture of the duodenum to one patient. RESULTS: The mortality was 0%. Came whereas the morbidity came basically on the seriousness of the accompanying injuries. PMID- 16355857 TI - [Reintervention in abdominal surgery. Personal experience in emergency]. AB - The Authors, regarding their emergency surgical experience, examine the need of reintervention in abdominal surgery at a distance (operations carried out not less than 30 days after previous procedure), a controversial situation, usually dealt with empirical approach. Such a feature is not rare in the experience of a surgeon; nevertheless still few are the studies concerning the severity of the disease and related complications requiring surgery. Occasionally it is really difficult--quite impossible- in these often complex clinical conditions, to identify the aetiology and even to perform a pre-operative diagnosis. Sometimes other factors may inhibit an exhaustive evaluation (acute onset of the symptoms, time spent from the former procedure, choice of the patient to refer to another surgeon) thus conditioning an adequate review of the first surgical act. The Authors study the more frequently observed clinical features, post-surgical abdominal adhesive syndrome, cancer recurrence, incisional hernia. Former surgical diseases and the latter one may be the same, but not necessarily. Surgery is only a feature of the treatment (multimodal treatment) in case of neoplastic recurrence. Morbidity and mortality concerning the latter surgical procedure are higher than the mere sum of those related to both the former and last operation performed (pre-operative disease understaging). Finally it must be stressed the need for early and accurate diagnosis to clearly steer the choice and course of surgical action. PMID- 16355858 TI - [The ambulatory treatment of varicose veins pathology. Personal experience]. AB - OBJECTIVE: In time of Surgery Cost Planning, progress is every step to ward an efficient and complete ambulatory treatment of pathology. In the field of venous surgery, a large share of patients take profit from a complete treatment in a single hospitalisation day. MATERIALS: The Authors carried out 21 strippings of great saphenous vein in 21 patients with local and regional anaesthesia. RESULTS: The results were the same as those of the traditional operation carried out with the help of spinal anaesthesia for a greater safety and comfort of the surgeons action. Moreover the authors obtained an immediate take off of the patient and his discharging after a few hours under observation. COMMENTS: The particular surgery technique perfectly copes with the anaesthetic method and fully profits from it because it limits local complication and unacceptable aesthetic defects. The chosen anaesthetic technique and the early take off haven't brought to any complications; moreover subcutaneous bleeding has been reduced, due to the absence of vasoparalysis typical of spinal anaesthesia. PMID- 16355859 TI - An original use in low colo-rectal anastomosis of the new surgical device "grasping tie". AB - The grasping tie is an original and mechanical tool, conceived and planned to be used in digestive surgery to do the equivalent of a purse string. Its use is conceived to make easier the mechanical anastomosis in the more difficult situations as the oesophago-jejunal or the very low colo-rectal ones, as in the example of the presented paper. A thin ribbon of nylon will be progressively tightened as a one-way running knot upon the tract of the gut, either directly or about the axis of a circular stapler. Once completed the stapler suture it will be completely extracted together with the muscle-mucosal rings, so that nothing of the ribbon remains in the patient. The use of the grasping tie is also advantageous in quickly closing the gut above and below a tumour, to a safer manipulation and to reduce the contamination of the operative field. An original and useful use of the grasping tie is demonstrated in the present paper for the surgical treatment of a low rectal cancer with preservation of the anus. Its use is that to close the distal stump of the rectum below the tumour, to take away from the pelvis the proximal stump with the tumour, and then to proceed to the mechanical anastomosis in a now wide operatory field, introducing in the anus an enteral dilator and preparing on it the definitive purse string, as illustrated. PMID- 16355860 TI - [A rare case of emphysematous gastritis by Salmonella arizonae]. AB - Gas within the wall of the stomach (emphysematous gastritis) is a rare condition associated with systemic toxicity and very often mortal end. The etiology includes firstly infections with gas-foaming organisms, other predisposing causes are the ingestion of corrosive substances, alcohol abuse and immunosuppression. But aplastic anemia hemodialysis even guzzling excessive amounts of carbonated beverage have being reported. Diagnosis is based on radiological techniques. There is no definitive treatment for emphysematous gastritis but the conservative approach with broad spectrum antibiotics seems always to be the first step. In this case, the guilty organism was Salmonella Arizonae. PMID- 16355861 TI - Giant aneurysm of the splenic artery in an elderly man short report and review of the literature. AB - Splenic Artery Aneurysms (SAAs) are usually single and small lesions, and their size rarely exceed 3 cm. In a review of the literature from 1950 to date, only 18 aneurysms defined as "giant" were found in 15 reported papers. CASE REPORT: A case of an 87-year-old man, successfully treated for a 7 cm wide aneurysm of the splenic artery is reported. Except for his age, the patient did not show any significant association with aneurysm-related diseases and was successfully submitted to en-bloc aneurysmectomy and splenectomy via open surgery. PMID- 16355862 TI - [Ulnar nerve pressure at the level of the elbow. Treatment by resection of the paratroclear apophysis]. AB - The pressure of the ulnar nerve at the level of the elbow is a pathological situation which causes serious problems of use of the suffering upper limb to its patient. In this study a reference is made to the anatomy of the ulnar nerve, to the reasons and to the main anatomic points of pressure of the ulnar nerve on the elbow, to the clinical indications and to the subjective symptoms of the patient. The state of art according to the diagnosis and methods of treatment is reported. The Authors particularly outline on the surgical technique used in order to solve this particular problem and to the patients that went to an under medical inspection. A step-by-step description of Authors' surgical technique and its benefits as well as its drawbacks is reported with an additionally, full iconography. PMID- 16355863 TI - [Informed consent in thyroid and parathyroid surgery]. AB - The Authors outline the law problems and the practice implications of thyroid and parathyroid surgery from the point of view of the informed consensus of the patient, and expose the criteria they usually adopt in their clinical practice for such procedures. This problem is particularly important because is exceptionally needed in emergency; therefore the preoperative information must be completed as possible, illustrating the risks of the procedure and the possible solutions alternative to surgery. PMID- 16355864 TI - [Legal contentions in thyroid surgery. Kind of preoperative information to the patients]. AB - The question of patients information and the achievement of an informed consent to medical and/or surgical treatment by the patient still represents one of the main issue of the medical profession, having greater importance nowadays in respect to the past regarding the relationship doctor-patient and to the diagnostic-therapeutic approach to the patient. Written informed consent is a pre requisite for surgical intervention as it provides the forum for the patient to appreciate implications of the procedure and the doctor to explain details and effects of the surgery. Patients refusal is, nowadays, according to the latest orientation of the Italian Supreme Court, the major limitation to medical or surgical intervention. For these reason we have thought better to propose a written form of informed consent regarding the thyroid surgery, which implies, more than other surgery's act, the exposition of the patient to adverse iatrogenic risks and as a consequence it can expose the surgeon to criminal or civil liabilities. PMID- 16355865 TI - Virus perpetuation in populations: biological variables that determine persistence or eradication. AB - In this review, I use the term "perpetuation" for persistence of a virus in a population, since this is a different phenomenon from persistence of a virus in an infected host. Important variables that influence perpetuation differ in small (<1000 individuals) and large (>10,000) populations: in small populations, two important variables are persistence in individuals, and turnover of the population, while in large populations important variables are transmissibility, generation time, and seasonality. In small populations, viruses such as poliovirus that cause acute infections cannot readily be perpetuated, in contrast to viruses such as hepatitis B virus, that cause persistent infections. However, small animal populations can turnover significantly each year, permitting the perpetuation of some viruses that cause acute infections. Large populations of humans are necessary for the perpetuation of acute viruses; for instance, measles required a population of 500,000 for perpetuation in the pre-measles vaccine era. Furthermore, if an acute virus, such as poliovirus, exhibits marked seasonality in large populations, then it may disappear during the seasonal trough, even in the presence of a large number of susceptible persons. Eradication is the converse of perpetuation and can be used as a definitive approach to the control of a viral disease, as in the instance of smallpox. Therefore, the requirements for perpetuation have significant implications for practical public health goals. PMID- 16355866 TI - The virus-immunity ecosystem. AB - The ecology of pathogenic viruses can be considered both in the context of survival in the macro-environments of nature, the theme pursued generally by epidemiologists, and in the micro-environments of the infected host. The long lived, complex, higher vertebrates have evolved specialized, adaptive immune systems designed to minimise the consequences of such parasitism. Through evolutionary time, the differential selective pressures exerted variously by the need for virus and host survival have shaped both the "one-host" viruses and vertebrate immunity. With the development of vaccines to protect us from many of our most familiar parasites, the most dangerous pathogens threatening us now tend to be those "emerging", or adventitious, infectious agents that sporadically enter human populations from avian or other wild-life reservoirs. Such incursions must, of course, have been happening through the millenia, and are likely to have led to the extraordinary diversity of recognition molecules, the breadth in effector functions, and the persistent memory that distinguishes the vertebrate, adaptive immune system from the innate response mechanisms that operate more widely through animal biology. Both are important to contemporary humans and, particularly in the period immediately following infection, we still rely heavily on an immediate response capacity, elements of which are shared with much simpler, and more primitive organisms. Perhaps we will now move forward to develop useful therapies that exploit, or mimic, such responses. At this stage, however, most of our hopes for minimizing the threat posed by viruses still focus on the manipulation of the more precisely targeted, adaptive immune system. PMID- 16355867 TI - Regulation of rodent-borne viruses in the natural host: implications for human disease. AB - Prevalence and transmission rates of rodent-borne viruses within host populations vary in time and space and among host-virus systems. Improving our understanding of the causes of these variations will lead to a better understanding of changes in disease risk to humans. The regulators of prevalence and transmission can be categorized into five major classes: (1) Environmental regulators such as weather and food supply affect transmission rates through their effect on reproductive success and population densities. (2) Anthropogenic factors, such as disturbance, may lead to ecosystem simplification and decreased diversity. These changes favor opportunistic species, which may serve as reservoirs for zoonotic viruses. (3) Genetic factors influence susceptibility of mice to infection or capacity for chronic shedding and may be related to population cycling. (4) Behavioral factors, such as fighting, increase risk of transmission of some viruses and result in different patterns of infection between male and female mice. Communal nesting may result in overwinter transmission in colder climates. (5) Physiologic factors control host response to infection and length of time the host remains infectious. Risk prediction is difficult because these regulators are numerous and often interact, and the relative importance of each varies according to the host species, season, year, and geographic location. PMID- 16355868 TI - Population dynamics of RNA viruses: the essential contribution of mutant spectra. AB - Cells and their viral and cellular parasites are genetically highly diverse, and their genomes contain signs of past and present variation and mobility. The great adaptive potential of viruses, conferred on them by high mutation rates and quasispecies dynamics, demands new strategies for viral disease prevention and control. This necessitates a more detailed knowledge of viral population structure and dynamics. Here we review studies with the important animal pathogen Foot-and-mouth disease virus (FMDV) that document modulating effects of the mutant spectra that compose viral populations. As a consequence of interactions within mutant spectra, enhanced mutagenesis may lead to viral extinction, and this is currently investigated as a new antiviral strategy, termed virus entry into error catastrophe. PMID- 16355869 TI - Pathogenesis of Rift Valley fever virus in mosquitoes--tracheal conduits & the basal lamina as an extra-cellular barrier. AB - Knowledge of the fate of an arbovirus in a mosquito is fundamental to understanding the mosquito's competence to transmit the virus. When a competent mosquito ingests viremic vertebrate blood, virus infects midgut epithelial cells and replicates, then disseminates to other tissues, including salivary glands and/or ovaries. The virus is then transmitted to the next vertebrate host horizontally via bite and/or vertically to the mosquito's offspring. Not all mosquitoes that ingest virus become infected or, if infected, transmit virus. Several "barriers" to arbovirus passage, and ultimately transmission, have been identified in incompetent or partially competent mosquitoes, including, among others, gut escape barriers and salivary gland infection barriers. The extra cellular basal lamina around the midgut epithelium and the basal lamina that surrounds the salivary glands may act as such barriers. Midgut basal lamina pore sizes are significantly smaller than arboviruses and ultrastructural evidence suggests that midgut tracheae and tracheoles may provide a means for viruses to circumvent this barrier. Further, immunocytochemical evidence indicates the existence of a salivary gland infection barrier in Anopheles stephensi. The basal lamina may prevent access to mosquito cell surface virus receptors and help explain why anopheline mosquitoes are relatively incompetent arbovirus transmitters when compared to culicines. PMID- 16355870 TI - The virulence of the 1918 pandemic influenza virus: unraveling the enigma. AB - The 1918 influenza pandemic caused acute illness in 25-30% of the world's population and resulted in the death of up to 40 million people. Using lung tissue of 1918 influenza victims, the complete genomic sequence of the 1918 influenza virus is being deduced. Neither the 1918 hemagglutinin nor neuraminidase genes possess mutations known to increase tissue tropicity that account for virulence of other influenza virus strains, such as A/WSN/33 or the highly pathogenic avian influenza H5 or H7 viruses. Using reverse genetics approaches, influenza virus constructs containing the 1918 hemagglutinin and neuraminidase on an A/WSN/33 virus background were lethal in mice. The genotypic basis of this virulence has not yet been elucidated. The complete sequence of the non-structural (NS) gene segment of the 1918 virus was deduced and also tested to determine the validity of the hypothesis that enhanced virulence in 1918 could have been due to type I interferon inhibition by the NS1 protein. Results from these experiments suggest that in human cells the 1918 NS1 is a very effective interferon antagonist. Sequence analysis of the 1918 influenza virus is allowing us to test hypotheses as to the origin and virulence of this strain. This information should help elucidate how pandemic influenza virus strains emerge and what genetic features contribute to virulence in humans. PMID- 16355871 TI - Transient or occult HIV infections may occur more frequently than progressive infections: changing the paradigm about HIV persistence. AB - Evidence of transient HIV infections was found in 8 subjects at high-risk for HIV infection among 47 longitudinally studied over 2-5 (average approximately 3.5) years, whereas only two subjects developed progressive infection. All of these subjects developed serum antibodies (Ab) to conformational epitopes of HIV gp41 (termed "early HIV Ab"), but the 8 transiently infected subjects lost this Ab within 4-18 months, and did not seroconvert to positivity in denatured antigen EIA or Western Blot (WB). However, the two progressively infected subjects eventually seroconverted in the EIA and WB tests within one to two months after the appearance of "early HIV Ab". HIV env and nef sequences were directly PCR amplified from the peripheral blood mononuclear cells (PBMCs) of two of the eight transiently infected subjects during the time of "early HIV Ab"-postivity, and these showed significant sequence divergence from the HIV strains in the laboratory, indicating that they were not laboratory contaminants. Genome identity typing ("paternity-typing") of PBMC samples obtained at the time of "early HIV Ab"-positivity, and later when Ab was absent from each of the 8 subjects, showed that blood samples were not mixed-up. This provides further evidence that transient or occult infection with HIV does occur, and perhaps at a greater frequency than do progressive infections. PMID- 16355872 TI - The role of reverse genetics systems in determining filovirus pathogenicity. AB - The family Filoviridae is comprised of two genera: Marburgvirus and Ebolavirus. To date minigenome systems have been developed for two Ebola viruses (Reston ebolavirus and Zaire ebolavirus [ZEBOV]) as well as for Lake Victoria marburgvirus, the sole member of the Marburgvirus genus. The use of these minigenome systems has helped characterize functions for many viral proteins in both genera and have provided valuable insight towards the development of an infectious clone system in the case of ZEBOV. The recent development of two such infectious clone systems for ZEBOV now allow effective strategies for experimental mutagenesis to study the biology and pathogenesis of one of the most lethal human pathogens. PMID- 16355873 TI - Species barriers in prion diseases--brief review. AB - Transmissible spongiform encephalopathies (TSEs or prion diseases) are neurological disorders associated with the aggregation of a pathologic isoform of a host-encoded protein, termed prion protein (PrP). The pathologic isoform of PrP, termed PrP(Sc), is a major constituent of the infectious agent. TSE diseases are characterized by neurodegenerative failure and inevitable morbidity. Bovine spongiform encephalopathy (BSE) has been transmitted from cattle to humans to cause a new variant of Creutzfeldt-Jakob syndrome. The potential for chronic wasting disease to similarly cross the species barrier from cervids to humans is considered unlikely but possible. Thus, understanding how TSE agents overcome resistance to transmission between species is crucial if we are to prevent future epidemics. The species barrier usually can be abrogated to varying degrees in laboratory animals. Studies done with transgenic animals, tissue culture, and cell-free assays established PrP as being necessary for TSE pathogenesis and illustrated that certain amino acid residues are more influential than others for conferring resistance to TSE agent transmission. The essence of what constitutes a TSE agent's species compatibility is thought to be orchestrated by a complex interplay of contributions from its primary amino acid sequence, its glycoform patterns, and its three-dimensional structure. PMID- 16355874 TI - Academic science and the business of vaccines. PMID- 16355875 TI - Emerging infectious diseases: the public's view of the problem and what should be expected from the public health community. PMID- 16355876 TI - [Concentration or network?]. PMID- 16355877 TI - [Is intrinsic or non-atopic asthma a particular disease?]. AB - Intrinsic or non-atopic asthma is defined by the absence of positive skin test for specific aero-allergens, together with the absence of specific serum IgE. In the adult population, about one third of asthmatic patients are non-atopic. Non atopic asthma begins generally later in life and is often more severe than atopic asthma. It is more often associated with chronic sinusitis and nasal polyps, a fact that is often underestimated. At the cellular and molecular level, non atopic asthma and atopic asthma a very similar, and the reason for their clinical difference is not really understood. PMID- 16355878 TI - [Immunization and immuno-modulation for prevention of respiratory tract infections]. AB - Respiratory tract infections are a major public health issue. Prevention in high risk populations relies mainly on vaccination against Influenza and S. pneumoniae. Vaccination of health-care workers is highly recommended, to decrease absenteism, but above all to protect high risk patients. New conjugate vaccines have shown their effectiveness in the paediatric population. In patients with chronic bronchitis or COPD, immunomodulatory agents (OM-85 BV) and anti-oxidants (NAC) are probably contributive in decreasing exacerbation rates. Inhaled corticosteroids decrease exacerbations in a well defined group of severe COPD. In patients with diffuse bronchiectasis, the immunomodulatory effect of macrolides, and the use of inhaled corticosteroids should be confirmed by larger clinical investigations. PMID- 16355879 TI - [Chronic thromboembolic pulmonary hypertension]. AB - Chronic thromboembolic pulmonary hypertension (CTEPH) is caused by obstruction of large pulmonary arteries by acute or recurrent emboli, organisation of these clots, and vascular remodeling of occluded as well as non-occluded peripheral arteries. Up to 4% of patients surviving from an acute embolic event will eventually develop chronic pulmonary hypertension. Major goals of the diagnostic work-up of pulmonary hypertension include the determination of its cause, the evaluation of its functional and haemodynamic repercussions, and if thromboembolic disease is present, the exact mapping of the pulmonary vascular bed obstruction. Pulmonary endarterectomy is the treatment of choice for selected patients. Therapeutic alternatives include lung or heart-lung transplantation, pulmonary angioplasty and pharmacological treatment with pulmonary vasodilators. PMID- 16355881 TI - [Pulmonary emphysema: mechanisms and therapeutic perspectives]. AB - Emphysema is one of the components of chronic obstructive pulmonary disease beside the bronchial, vascular and systemic parts. The pathogenesis involves an inflammatory process (macrophages, neutrophiles and lymphocytes), but also an imbalance between proteases-anti-proteases and the oxidative stress by the formation of oxygen radicals. On the therapeutic level, these physiopathological pathways are the rational basis to the progressive use of antioxidants (mainly N acetylcysteine) or, for the moment at an experimental level, of anti-proteases (inhibitors of serine proteases, cysteine proteases and the matrix metalloproteases). To reach the secondary lobule, site of the emphysema, the inhaled future treatments will have to be constituted of small particles, as in cigarette smoke, in order to distribute into the most homogeneous way the affected lung. PMID- 16355880 TI - [Hemoptysis: methods of localization]. AB - Depending on the amount of bleeding, chest radiograph localises the origin of an hemoptysis in 20 to 50% of cases. Computed tomography (CT) scan of the chest is the most accurate method used to localise and identify the source of bleeding. In case of normal imaging, bronchoscopy localises the bleeding source in 40% of cases. Bronchogenic carcinoma was identified in 3% of the bronchoscopies performed for hemoptysis in patients with a normal chest roentgenogram. The evaluation of a minor hemoptysis in a patient without risk factor for a lung cancer could be limited to a CT-scan of the chest if this one is normal. If the prognosis of an idiopathic hemoptysis is generally good, the presence of a smoking history and an age over 50 years justifies a radiologic follow up to exclude a growing bronchial tumor. PMID- 16355882 TI - [Do inhaled corticosteroids reduce mortality in chronic obstructive pulmonary disease?]. AB - In Switzerland, high dose of inhaled corticosteroids are frequently prescribed to COPD patients. Observational studies have suggested a reduction in mortality, although several randomised controlled trials did not show such a beneficial effect. Two important exposure biases are highlighted by a careful review of the methodological aspects of these observational studies: the immortal time bias and the failure to perform a time dependent analysis lead to underestimate the risk of death among exposed patients therefore incorrectly suggesting a survival benefit of regular inhaled steroid use. Two recent pharmaco-epidemiological studies, controlling for these exposure biases did not find any reduction in mortality due to inhaled steroids among COPD patients. PMID- 16355883 TI - [Extrapulmonary effects of chronic obstructive pulmonary disease]. AB - Patients suffering from advanced COPD not only present respiratory symptoms but extrapulmonary manifestations as well: muscle dysfunction, weight loss, osteoporosis, sexual dysfunction, cardiovascular diseases. A chronic inflammation is likely to play a role in the occurrence of these associated disorders. These different aspects contribute to a poor quality of life, and to added morbidity and mortality. The presence of systemic effects in COPD means that the patient suffering from this pulmonary disease needs a comprehensive care. PMID- 16355884 TI - [Mirror, mirror on the wall... Or the right to contract presented as a fact account]. PMID- 16355885 TI - [Practical problems of the ECG in the elderly. Case No. 4]. PMID- 16355886 TI - [On the eve of the Geneva Conference]. PMID- 16355887 TI - [The Rylander affair: science corrupted knowingly]. PMID- 16355888 TI - [Tamiflu: 300 million treatments will be produced in the next few months]. PMID- 16355889 TI - [November 2005, France and her "scum"]. PMID- 16355890 TI - [Mucoviscidosis: France is heading towards biological rather than genetic screening]. PMID- 16355891 TI - [Bariatric surgery explodes in the United States]. PMID- 16355893 TI - The pet prescription. PMID- 16355892 TI - [Maieutics]. PMID- 16355894 TI - Hurricane Katrina: now the real work begins. PMID- 16355895 TI - Are your hospital reports online yet? PMID- 16355896 TI - Why these docters love their RHIO. PMID- 16355897 TI - Is a tablet right for you? PMID- 16355898 TI - Well-woman exams. PMID- 16355899 TI - Fire the transcriptionist--voice recognition works. PMID- 16355901 TI - ICD-9: what's changed. PMID- 16355900 TI - How young doctors are doing today. PMID- 16355902 TI - Caring for refugees. PMID- 16355903 TI - Don't bother donating expired drugs. PMID- 16355904 TI - Where does your state rank? PMID- 16355905 TI - If the patient needs a consult. PMID- 16355906 TI - When you refer the patient. PMID- 16355907 TI - How to respond to nasty letters. PMID- 16355908 TI - Practice mergers: what makes a winner. PMID- 16355909 TI - Secret weapons for a successful practice. PMID- 16355910 TI - If you cover for post-op patients. PMID- 16355911 TI - Autonomy in physical therapy: less is more. PMID- 16355912 TI - Primary care physical therapy practice models. AB - The purpose of this paper is to provide a brief background on the concept of primary care physical therapy, describe 3 existing models of primary care physical therapy, explore their similarities and differences, and discuss the potential implications and opportunities for the profession. The programs at US Army medical facilities, Kaiser Permanente Northern California, and the Department of Veterans Affairs Salt Lake City Health Care System are presented by the author affiliated with each respective program. PMID- 16355913 TI - Musculoskeletal imaging in physical therapist practice. AB - This article presents an overview of current concepts of evidence-based diagnosis using a variety of imaging modalities for a broad spectrum of musculoskeletal conditions and syndromes. There is limited but increasing evidence that physical therapists appropriately use diagnostic studies in clinical practice. Pathology revealed by diagnostic studies must be viewed in the context of the complete examination, as pathology is common in the asymptomatic population. Special diagnostic challenges are presented by patients with areas of referred pain, multiple injuries or multiple areas of pathology, neoplasms, and infections. Plain film radiographs have been overused in the clinical management of many conditions, including low back pain. Clinical decision rules provide simple evidence-based guidelines for the appropriate use of imaging studies. PMID- 16355914 TI - Cervical cord compression secondary to ossification of the posterior longitudinal ligament. AB - STUDY DESIGN: Resident's case problem. BACKGROUND: A 52-year-old Chinese male with a 10-year history of gradually worsening right hip stiffness, weakness, and pain was referred to physical therapy by his orthopedist, who made a diagnosis of developmental dysplasia of the right hip, with possible Legg-Calve-Perthes disease. The patient reported multiple falls over the last several years and a gradual onset of low back pain with an onset of "electricity" down both legs. The patient also reported mild numbness in both forearms and the right hand over the previous several months. This resident's case problem illustrates how a physical therapist recognized the presence of an atypical musculoskeletal pathology through the use of hypothesis-driven clinical reasoning and detailed physical examination. DIAGNOSIS: Examination of the patient's lumbar and cervical spine and hips revealed joint dysfunctions. Neurological testing revealed hyperreflexia. Special testing revealed lower extremity clonus with a positive Babinski sign with gait disturbances. The patient was referred back to his primary physician and then to a neurologist and neurosurgeon. An MRI revealed cervical myelopathy due to ossification of the posterior longitudinal ligament from C3/C4 to C5/C6. The patient then underwent a C3 through C7 laminectomy. DISCUSSION: It is always imperative that sound clinical reasoning be used when performing physical therapy evaluations, regardless of the referral status of the patient. Patients with nonmusculoskeletal pathology may seek physical therapy services and it is the physical therapist's responsibility to complete a thorough examination and refer to specialists when appropriate. PMID- 16355915 TI - Cardiovascular assessment in the orthopaedic practice setting. AB - As consumer access to physical therapy practice expands, it is important that physical therapists are familiar with and implementing accepted methods of identifying the cardiovascular status of their clients. Established guidelines for assessing cardiovascular risk prior to initiating aerobic exercise programs are available and can be readily adopted by physical therapists in diverse clinical settings. We have provided a process for integrating existing guidelines into clinical practice. Because little evidence exists regarding the clinical behaviors and knowledge of orthopedic physical therapists in the area of cardiovascular risk, we conducted a survey to assess current practice patterns. The results suggest that orthopedic physical therapists are performing cardiovascular screening at frequencies similar to other components of the history and systems review, but that monitoring baseline or exercising vital signs does not occur with every exercise session. PMID- 16355916 TI - Decision making for a painful hip: a case requiring referral. AB - STUDY DESIGN: Resident's case problem. BACKGROUND: The purpose of this resident's case problem is to describe a 39-year-old female patient with insidious onset of hip pain. This patient had discrete findings on subjective physical examination that prompted referral for further imaging studies of the left hip and pelvis. Despite having seen multiple providers, no imaging of the involved hip or pelvis had been performed. A prolonged duration of symptoms, severe gait disturbance with an associted Trendelenburg sign, difficulty sleeping, and an empty end feel with passive range of motion increased concern that a pathological process might be present. DIAGNOSIS: Imaging studies revealed a large destructive soft-tissue tumor later found to be non-Hodgkin's lymphoma. DISCUSSION: It is incumbent upon physical therapists to be aware of the potential for severe pathological conditions that mimic musculoskeletal complaints to exist and understand how to identify patients for whom further testing and/or referral may be appropriate. Existing guidelines for low back pain may assist with decision making in the absence of specific guidelines for when to request imaging in patients with nontraumatic hip and pelvis pain. Proficiency in screening for conditions not amenable to physical therapy treatment or that require consultation to other health care professionals is essential to physical therapy practice. PMID- 16355917 TI - Diagnostic imaging and differential diagnosis in 2 case reports. AB - STUDY DESIGN: Retrospective resident's case reports. BACKGROUND: In today's healthcare setting, it is important for physical therapists to recognize when diagnostic imaging is necessary--as well as know how to interpret the results of these tests--to assist in the clinical decision-making process. Two cases are presented that illustrate how a physical therapist, credentialed to request and review diagnostic imaging, effectively and efficiently utilized multiple forms of diagnostic imaging to assist in his differential diagnosis and clinical decision making. DIAGNOSIS: The first case report describes the differential diagnostic process for a 33-year-old active duty military paratrooper who had sustained trauma to his neck. His history was consistent with a C6 radiculopathy, which was confirmed by a neurological screening examination. Radiographs requested by the physical therapist revealed an anterolithesis of C5 on C6, with a possible fracture. An orthopedic surgeon was consulted and further diagnostic testing via magnetic resonance imaging revealed a large disc herniation at C5-6, with spinal cord compression, as well as a C5 vertebral body fracture with nearly perched facets at C5 on C6. The patient was subsequently referred to a neurosurgeon and underwent an emergency C5-6 fusion that afternoon. The second case report describes the differential diagnosis of a 20-year-old active-duty soldier referred for rehabilitation with a diagnosis of a distal fibula stress fracture. Previous treatment by the referring provider included 3 months of rest and anti inflammatory medications. Physical examination of the patient revealed a marked decrease in ankle inversion with a firm end feel. This was not consistent with the diagnosis established by the referring provider. Subsequent radiographs requested by the physical therapist and a computed tomography scan requested by a podiatrist revealed synostosis of the middle facet of the talocalcaneal joint with an apparent fracture line. The patient subsequently underwent a subtalar arthrodesis. DISCUSSION: In these cases the physical therapist requested imaging needed for appropriate management, despite the patient having previously seen a primary care provider. In both examples, the physical therapist successfully identified abnormalities prior to a radiologist or other physician reviewing the results. This avoided delay in definitive management of the patients' problems. It is imperative that physical therapists understand when diagnostic imaging is necessary to assist in the differential diagnosis of patients. Likewise, it is important for physical therapists to be competent in interpreting the results of these tests. When not in a direct access physical therapy environment, a physical therapist should understand when diagnostic imaging tests are indicated. This facilitates working with the entire health care team to acquire necessary tests in an appropriate timeframe. PMID- 16355918 TI - Abdominal differential diagnosis in a patient referred to a physical therapy clinic for low back pain. AB - STUDY DESIGN: Resident's case problem. BACKGROUND: Acute back pain most often presents as musculoskeletal in nature; however, less frequently it may be the result of an underlying, or coexisting, systemic pathology. When present, the signs and symptoms of systemic pathology can mimic, or be masked by, musculoskeletal back pain, which may pose a diagnostic challenge during the clinical evaluation. The purpose of this resident's case problem is to describe the clinical reasoning process leading to a medical referral for a patient who presented to physical therapy with debilitating low back pain. DIAGNOSIS: The patient in this resident's case problem was a 67-year-old male referred to physical therapy with a 2-week history of severe low back pain and muscle spasms. The patient history and physical examination were suggestive of musculoskeletal back pain and physical therapy treatment was initiated. Abdominal pain was elicited during an introductory therapeutic exercise, which was recognized by the therapist as a potential sign of abdominal pathology. The therapist performed an additional review of systems and an abdominal screening examination, which established the necessity of an immediate medical referral. At the emergency department, ominous abdominal pathology was safely ruled out through diagnostic imaging and the patient was treated for secondary gastrointestinal effects of opioid analgesic medications. DISCUSSION: This resident's case problem provides an opportunity to discuss the clinical reasoning process leading to the suspicion of abdominal pathology. Specifically, this case reinforces the importance of recognizing potential signs of systemic pathology, executing an appropriate physical examination, including screening of the involved anatomical region, and providing an appropriate medical referral when indicated. PMID- 16355919 TI - BNP as a biomarker in heart disease. AB - Heart ventricles produce B-type natriuretic peptide (BNP) in response to increased mechanical load and wall stretch. BNP protects the heart from adverse consequences of overload by increasing natriuresis and diuresis, relaxing vascular smooth muscle, inhibiting the renin-angiotensin-aldosterone system, and by counteracting cardiac hypertrophy and fibrosis. BNP is synthesized by human cardiac myocytes as a 108-amino acid prohormone (proBNP), which is cleaved to the 32-residue BNP and the 76-residue N-terminal fragment of proBNP (NT-proBNP). Both can be used as sensitive biomarkers of cardiac dysfunction and well-characterized commercial assays have recently become available. In acute coronary syndromes increased concentrations are strong predictors of recurring myocardial infarction, heart failure, and death. In acute dyspnea, high BNP and NT-proBNP point to a cardiac rather than a pulmonary origin of the symptoms. BNP and NT proBNP help in the assessment of the severity of ventricular dysfunction and heart failure and as a prognostic predictor, regardless of the primary cause of the condition. They can be used to guide the therapy of heart failure and left ventricular dysfunction. BNP and NT-proBNP work better when they are used for specific clinical purposes, rather than for screening in the general population. Their main strength is the excellent negative predictive value with regard to left ventricular dysfunction and heart failure. BNP and NT-proBNP are nonspecific biomarkers of cardiac dysfunction. Specific diagnostic tools, such as echocardiography, are required to define the actual abnormality. PMID- 16355920 TI - Pathways of myocyte death: implications for development of clinical laboratory biomarkers. AB - The recognition that cardiac myocytes die by multiple mechanisms and thus substantially affect ventricular remodeling in diseased human hearts supports the concept of ongoing myocyte death in the progression of heart failure and constitutes the basis of this review. In addition, based on the pathophysiology of myocardial cell deaths, the present study emphasizes that currently methodologies, although with some inherent limitations, are available to recognize and measure quantitatively the contribution of myocyte cell death to the progression of the pathologic state of the heart. Our own studies show that application of such methodologies including modern microscopy techniques and the use of different molecular and immunohistochemical markers may generate the consensus that myocyte cell death is a quantifiable parameter in the normal and pathological human heart. The present study also demonstrates that myocyte cell death, apoptotic, oncotic or autophagic in nature, has to be regarded as an additional critical variable of the multifactorial events implicated in the alterations of cardiac anatomy and myocardial structure of the diseased human heart. PMID- 16355921 TI - Breast cancer biomarkers. AB - Substantial progress has been made over the past three decades in our understanding of the epidemiology, clinical course and basic biology of breast cancer. This chapter considers the existing ancillary tests and emerging molecular markers in breast cancer prognosis assessment and the prediction of response of breast cancer to treatment of the disease. PMID- 16355922 TI - Autoantibodies in autoimmune liver disease. AB - Autoantibodies indicate an immune reactive state, but in liver disease they lack pathogenicity and disease specificity. Antinuclear antibodies, smooth muscle antibodies, antibodies to liver/kidney microsome type 1, antimitochondrial antibodies, and perinuclear antineutrophil cytoplasmic antibodies constitute the standard serological repertoire that should be assessed in all liver diseases of undetermined cause. Antibodies to soluble liver antigen/liver pancreas, asialoglycoprotein receptor, actin, liver cytosol type 1, nuclear antigens specific to primary biliary cirrhosis, and pore complex antigens constitute an investigational repertoire that promises to have prognostic and diagnostic value. These autoantibodies may emerge as predictors of treatment response and outcome. Antibodies to histones, doubled-stranded DNA, chromatin, and lactoferrin constitute a supplemental repertoire, and they support the immune nature of the liver disease. Final diagnoses and treatment strategies do not depend solely on serological markers. Autoantibodies are floating variables, and their behavior does not correlate closely with disease activity. There are no minimum levels of significant seropositivity, especially in children. Over-interpretation is the major pitfall in the clinical application of the serological results. New autoantibodies will emerge as the search for target antigens and key pathogenic pathways continues. PMID- 16355923 TI - Mass spectrometry of advanced glycation end products. AB - Mass spectrometry, in particular matrix assisted laser desorption/ionisation, is a powerful analytical tool in studies devoted to protein non-enzymatic glycation. It has been firstly tested on in vitro glycated proteins, and looking at the reliable results so obtained, on in vivo glycated proteins in population of healthy, well-controlled and badly controlled diabetic patients. The comparison of the data so obtained in case of human serum albumin and IgG unequivocally demonstrates the highest glycation level for the third set of subjects. Further results obtained in the case of hemoglobin glycation showed that both alpha and beta globins are glycated in a similar extent and that the method can be employed to investigate on the "oxidative stress" experimented by the patients. PMID- 16355924 TI - Advances in real-time PCR: application to clinical laboratory diagnostics. AB - The polymerase chain reaction (PCR) has become one of the most important tools in molecular diagnostics, providing exquisite sensitivity and specificity for detection of nucleic acid targets. Real-time monitoring of PCR has simplified and accelerated PCR laboratory procedures and has increased information obtained from specimens including routine quantification and differentiation of amplification products. Clinical diagnostic applications and uses of real-time PCR are growing exponentially, real-time PCR is rapidly replacing traditional PCR, and new diagnostic uses likely will emerge. This review analyzes the scope of present and potential future clinical diagnostic applications of this powerful technique. Critical discussions focus on basic concepts, variations, data analysis, instrument platforms, signal detection formats, sample collection, assay design, and execution of real-time PCR. PMID- 16355925 TI - Growth factors in lung development. AB - Organized and coordinated lung development follows transcriptional regulation of a complex set of cell-cell and cell-matrix interactions resulting in a blood-gas interface ready for physiologic gas exchange at birth. Transcription factors, growth factors, and various other signaling molecules regulate epithelial mesenchymal interactions by paracrine and autocrine mechanisms. Transcriptional control at the earliest stages of lung development results in cell differentiation and cell commitment in the primitive lung bud, in essence setting up a framework for pattern formation and branching morphogenesis. Branching morphogenesis results in the formation of the conductive airway system, which is critical for alveolization. Lung development is influenced at all stages by spatial and temporal distribution of various signaling molecules and their receptors and also by the positive and negative control of signaling by paracrine, autocrine, and endocrine mechanisms. Lung bud formation, cell differentiation, and its interaction with the splanchnic mesoderm are regulated by HNF-3beta, Shh, Nkx2.1, HNF-3/Forkhead homolog-8 (HFH-8), Gli, and GATA transcription factors. HNF-3beta regulates Nkx2.1, a transcription factor critical to the formation of distal pulmonary structures. Nkx2.1 regulates surfactant protein genes that are important for the development of alveolar stability at birth. Shh, produced by the foregut endoderm, regulates lung morphogenesis signaling through Gli genes expressed in the mesenchyme. FGF10, produced by the mesoderm, regulates branching morphogenesis via its receptors on the lung epithelium. Alveolization and formation of the capillary network are influenced by various factors that include PDGF, vascular endothelial growth factor (VEGF), and retinoic acid. Epithelial-endothelial interactions during lung development are important in establishing a functional blood-gas interface. The effects of various growth factors on lung development have been demonstrated by gain- or loss-of-function studies in null mutant and transgenic mice models. Understanding the role of growth factors and various other signaling molecules and their cellular interactions in lung development will provide us with new insights into the pathogenesis of bronchopulmonary dysplasia and disorders of lung morphogenesis. PMID- 16355926 TI - Clinical laboratory evaluation of male subfertility. AB - Male subfertility is a common problem with a complex etiology, requiring a complete andrological work-up for proper diagnosis. The male reproductive tract is controlled by a well-balanced hormonal system, in which hypothalamic (GnRH), pituitary (LH, FSH) and testicular hormones (androgens, inhibin B) participate. Any disturbance of this hormonal system may therefore lead to testicular dysfunction and interfere with the spermatogenesis process. In addition, also other components along the ductal system, such as epididymis, prostate and seminal vesicles, that improve sperm fertility by contributing their secretions to the semen, might function inadequately and thus fail to enhance the fertilizing capacity of the sperm cells. External factors (heat, chemicals, life style) and anatomical abnormalities (varicocele) were shown to have a negative influence on male fertility. In a number of patients genetic defects can be identified as the cause of their infertility. Laboratory tests are available to assess hormone concentrations, semen composition, accessory gland function and sperm cell function. Conventional semen analysis includes the determination of sperm concentration, semen volume, sperm motility (qualitative and quantitative), sperm morphology, sperm cell vitality, pH, leucocytes and antibodies. The usefulness of the determination of these parameters as predictor of fertility appears to be rather limited, however. Therefore, alternative tests, some based on more functional aspects (sperm penetration, capacitation, acrosome reaction), have been developed. Furthermore, there is an increasing attention for the assessment of DNA integrity, for instance by the flowcytometer-based Sperm Chromation Structure Assay (SCSA), as an additional or alternative parameter of sperm quality. It is likely and desirable that further assays with better predictive value are being developed in the near future. PMID- 16355927 TI - What's new in MR safety: the latest on the safe use of equipment in the magnetic resonance environment. AB - Magnetic resonance (MR) scanners present a unique set of safety risks for the healthcare facility. In the space surrounding an MR scanner, ferromagnetic objects can become deadly projectiles, device cables can become hot enough to cause burns, and medical equipment can cease to function properly. ECRI detailed these risks--and provided recommendations to help healthcare facilities minimize them--in a December 2001 Guidance Article. In this supplement to that article, we detail some recent MR safety developments and offer additional guidance. In particular, we describe changes in the terminology used to characterize the safety of devices in the MR environment, we outline new guidance on MR safe practices published by the American College of Radiology, and we review the safety issues that facilities need to be aware of when implementing 3-tesla (3 T) MR systems. We also present an updated Starter List of medical device models that have been designed for use in the MR environment. In addition, supplementary articles address the use of ferromagnetic detection systems and the selection of a fire extinguisher for use in the MR environment. PMID- 16355928 TI - Newborn hearing screening devices: sound advice on choosing the right technology. AB - Infant hearing loss occurs more often than any other problem screened for at birth. Finding such hearing loss helps to prevent early developmental delays that can hinder a child's physical, mental, and emotional growth. To address this public health concern, many hospitals are implementing universal newborn hearing screening (UNHS) programs. They are called "universal" because the objective is to test all newborns. Such programs use auditory brainstem response (ABR), otoacoustic emissions (OAE), or a combination of the two to screen newborn hearing. A goal of any newborn hearing screening program is to reduce the false positive rates that result in unnecessary referrals. Such rates should be kept low because unnecessary referrals mean personnel must spend extra time scheduling follow-up testing, thus increasing program costs. Therefore, referral rates and costs will be the main factors when hospitals are choosing the appropriate screening technology. PMID- 16355929 TI - Misconnection between St. Jude Medical Pacel bipolar pacing catheter and pacing cable causes improper pacing. PMID- 16355930 TI - Sharing the stories of perioperative nursing. PMID- 16355931 TI - Perioperative nurse week: recognition of excellence. PMID- 16355932 TI - Vaginal prep solutions. PMID- 16355933 TI - Differing views on perioperative prima donnas. PMID- 16355934 TI - Differing views on perioperative prima donnas. PMID- 16355935 TI - Using an algorithm to easily interpret basic cardiac rhythms. AB - MANY NURSES STRUGGLE with identifying electrocardiogram (ECG) rhythms, but rapidly interpreting primary ECG rhythms is an essential skill that every nurse should master. THIS ARTICLE PROVIDES an algorithm that nurses can use to easily interpret basic ECG rhythms. PMID- 16355936 TI - Substance abuse among nurses--intercession and intervention. AB - IT IS NOT EASY to report a coworker who may have a substance abuse problem, so many nurses choose to remain silent about this issue. THIS ARTICLE PROVIDES suggestions for staff nurses about how to confront a peer, document inappropriate nursing behaviors related to substance abuse, and report these issues to a manager. The manager's role in counseling and intercession with a substance abusing employee also is detailed. REMEDIATION AND SUPPORT programs are addressed along with return-to-work issues and the need for education about this debilitating disease. PMID- 16355937 TI - Decreasing latitude and increasing regulation in transplantable tissue programs. AB - ADVANCED TECHNOLOGY and improved surgical techniques have led to new therapeutic uses for allografts. DISEASE TRANSMISSION via allograft tissue transplants has prompted federal intervention in the tissue banking industry and resulted in federal regulations. NEW STANDARDS from the Joint Commission on Accreditation of Healthcare Organizations became effective July 1, 2005, and apply to all hospitals that store or implant allograft tissues. These standards include mandatory policies on all aspects of hospital transplantation programs, including tissue ordering, receipt, storage, issuance, and record keeping. PMID- 16355938 TI - Anterior lumbar interbody fusion--advances in spinal fusion technology. AB - AS MUCH AS 80% OF THE US POPULATION will be affected by back pain at some time during their lives. Some of the most common disorders are herniated disc, degenerative disc disease, degenerative spondylolisthesis, spinal stenosis, and revision of previously failed low back surgery. IF CONSERVATIVE TREATMENT for back pain fails, spinal fusion may be performed. Anterior lumbar interbody fusion effectively manages degenerative joint disease, instability, and spondylolisthesis. NURSES WHO PROVIDE CARE for patients undergoing this procedure help ensure patient safety and promote positive outcomes. PMID- 16355939 TI - An emerging role for nurse practitioners--preoperative assessment. AB - NURSE PRACTITIONERS (NPs) are expanding their role beyond primary care, and preoperative assessment is one area in which existing medical and nursing models of care could be merged to create a specialty role for NPs. THIS ARTICLE REVIEWS health-related literature regarding the impact of NPs in the preoperative assessment setting and explores the potential for this emerging role as an alternative to existing preoperative assessment models. PMID- 16355940 TI - Using the DISC behavioral instrument to guide leadership and communication. AB - BEHAVIORAL INSTRUMENTS can be useful tools to help leaders gain insight into how to better communicate with coworkers. The DISC instrument classifies behaviors into four personality types (ie, Dominant, Influencer, Steady, Conscientious) and provides methods leaders can use to work with each personality type. LEADERS SHOULD STRIVE to have a good mix of personalities on their teams to achieve optimal success. THIS ARTICLE DESCRIBES the benefits of using the DISC behavioral evaluation method to better understand and work with team members and gives role play scenarios for dealing with each personality. PMID- 16355941 TI - The expert nurse witness. PMID- 16355942 TI - Antibiotic prophylaxis: what's the fuss? PMID- 16355943 TI - Sweetwater Hospital Association v Carpenter. PMID- 16355944 TI - Effects of preincisional ketamine treatment on natural killer cell activity and postoperative pain management after oral maxillofacial surgery. AB - Poorly controlled pain may lead to increased risk of cancer metastasis by suppressing natural killer (NK) cell activity. Ketamine may be beneficial by potentiating opioid-induced analgesia. We enrolled 59 participants in a randomized double-blind, placebo-controlled clinical trial and assigned them to receive propofol plus (1) saline, 2 mL; (2) ketamine, 0.5 mg/kg; or (3) ketamine, 1.2 mg/kg, followed by a standardized anesthesia protocol. The visual analogue scale (VAS) and 24-hour opioid consumption measured postoperative pain perception. NK cell activity was measured before and 24 hours after ketamine administration using the chromium 51 release assay. Nonparametric analysis of VAS data revealed that women receiving 0.5 mg/kg of ketamine reported less pain (P <.05) compared with the saline 1.2 mg/kg-ketamine groups. This finding was not evident in men. Comparing opioid consumption among the 3 groups (using analysis of variance) revealed a drug-gender interaction (P < .05): 0.5 mg/kg of ketamine decreased postoperative opioid consumption for women more than for men. Although not statistically significant, women receiving 0.5 mg/kg of ketamine had the least NK cell suppression compared with preoperative values (repeated analysis of variance). These findings suggest that for women, low-dose ketamine may be beneficial. PMID- 16355945 TI - Hyperkalemia-induced residual neuromuscular blockade: a case report. AB - Muscle relaxants are routinely administered during anesthesia. The residual effects of nondepolarizing muscle relaxants must be reversed by the anesthesia provider prior to extubation of the trachea to ensure that patients can safely protect their own airways and maintain adequate ventilation. Many factors can potentiate the effects of muscle relaxants, and each of these factors must be taken into consideration when postoperative muscle weakness is encountered. Only after the cause of residual neuromuscular weakness is discerned can the appropriate treatment be determined. This article describes the anesthetic management of a patient who experienced postoperative residual muscle weakness secondary to unsuspected hyperkalemia. A discussion of potassium's role in membrane excitability and factors that must be considered in the differential diagnosis of residual neuromuscular weakness is included. PMID- 16355946 TI - A study of occupational stress, scope of practice, and collaboration in nurse anesthetists practicing in anesthesia care team settings. AB - This study examined occupational stress in Certified Registered Nurse Anesthetists (CRNAs) practicing with anesthesiologists in anesthesia care team (ACT) settings. The focus was to examine the relationships among CRNA scope of practice (SOP) in ACTs, collaboration, and role-related occupational stress. A survey questionnaire was mailed to CRNAs from the 6 New England states, with a return rate of 30.87% (n = 347). Data analysis included practice characteristics and demographics of the sample, and the research questions were examined applying correlational analysis, t test, and analysis of variance addressing relationships among the study measures. Data analyses revealed that limited, restricted CRNA practice scope was particularly evident in respondents employed by anesthesiology groups, compared with hospital-employed CRNAs. Few CRNA respondents perceived their practice as collaborative, and many used compromise as a conflict resolution style. Respondents with a broader SOP reported higher collaboration than those with restrictions. Respondents reporting a broader SOP also experienced increased job stress in relation to role overload but used coping resources effectively. Implications for future studies include exploring strategies that achieve consensus between CRNAs and anesthesiologists in ACTs, emphasizing clearly defined roles and optimizing productivity. PMID- 16355947 TI - Update for nurse anesthetists--anesthetic considerations for patients with amyloidosis. AB - Amyloidosis is a rare disease process that results in the deposition of insoluble, fibrous amyloid proteins in extracellular spaces and tissues. Amyloid fibrils can be deposited locally or may involve every organ system of the body. Advancements in the treatment for amyloidosis allow longer survival, and patients are being seen in our operating rooms for diagnostic, interventional, and curative purposes. Amyloidosis has numerous implications for anesthesia providers due to the possibility of systemic involvement. This course describes 2 cases of amyloidosis and discusses the types of amyloidosis and their anesthetic implications. PMID- 16355948 TI - Long-term results of external dacryocystorhinostomy. AB - BACKGROUND AND OBJECTIVE: To evaluate the long-term success following external dacryocystorhinostomy, which is used as a standard in comparing endoscopic or laser techniques. PATIENTS AND METHODS: A chart review of 128 patients (150 procedures) undergoing external dacryocystorhinostomy within a 14-year period was undertaken. Patient demographics, symptoms, examination findings, surgical and anesthesia information, postoperative course, and complications were recorded. Lack of tearing or evidence of dacryocystitis at the final postoperative visit was considered a successful result. RESULTS: External dacryocystorhinostomy was found to be highly successful, with an overall 93% success rate at an average follow-up of 2.7 years. Kaplan-Meier analysis of the data, applied for the first time in this study, predicts a success rate of 90% beyond 4 years. Most important, this demonstrated that most surgical failures were identified in the first 2 years. Postoperative complications were rare and the surgical scar was not a source of patient concern in most cases. CONCLUSIONS: This study confirms that external dacryocystorhinostomy is a highly successful long-term solution to nasolacrimal duct obstruction with low morbidity. PMID- 16355949 TI - Early and late effects of pterygium surgery on corneal topography. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to compare the corneal topographic changes at the early and late postoperative period after pterygium surgery. PATIENTS AND METHODS: Sixteen eyes of 15 patients with primary pterygium were included in the study. All of the pterygia were excised primarily. Computerized corneal topography measurements were taken before surgery and at the second week (postoperative early period) and third month (postoperative late period) after the operation. Keratometric astigmatism, total mean corneal refractive power, and flattened or steepened areas of the cornea were detected at each visit. The topographical changes were evaluated using the analysis of variance. Multiple comparison analyses were made with the Tukey HSD test. RESULTS: Keratometric astigmatism was 2.58 +/- 2.33 diopters (D) at the preoperative period, 1.30 +/- 1.24 D at the postoperative early period, and 0.61 +/- 0.38 D at the postoperative late period (P < .001). The surgically induced corneal astigmatism was 3.23 +/- 2.33 D (range, 0.52 to 8.32 D) at the postoperative early period and 2.70 +/- 2.24 D (range, 0.47 to 7.72 D) at the postoperative late period (P = .287). Astigmatism was 81.25% with-the-rule at the preoperative period, 43.75% oblique and 37.50% against-the-rule at the postoperative early period, and 62.50% with-the-rule at the postoperative late period. Total mean corneal refractive power was 42.51 +/- 1.99 D at the preoperative period, 43.95 +/- 1.58 D at the postoperative early period, and 43.89 +/- 1.80 D at the postoperative late period (P = .014). CONCLUSION: Corneal topographic changes induced by pterygium greatly improve at the postoperative early period. However, normal corneal topography patterns are formed at the postoperative late period. PMID- 16355950 TI - Effect of apraclonidine versus dorzolamide on intraocular pressure after phacoemulsification. AB - BACKGROUND AND OBJECTIVE: To investigate the efficacy of 1% apraclonidine hydrochloride and 2% dorzolamide hydrochloride in controlling postoperative intraocular pressure elevation in patients undergoing phacoemulsification. PATIENTS AND METHODS: A prospective, randomized, double-blind, placebo-controlled study was conducted. Seventy-five patients were randomized to receive either 1% apraclonidine hydrochloride, 2% dorzolamide hydrochloride, or artificial tears immediately after uncomplicated phacoemulsification. Intraocular pressure was measured by Goldmann applanation tonometer 1 to 3 hours preoperatively and 24 hours and 5 days postoperatively. RESULTS: Mean intraocular pressure measured at 24 hours and 5 days after surgery was not significantly different between groups. None of the patients had an elevated intraocular pressure of 30 mm Hg or higher. No marked difference was observed in patient age, preoperative intraocular pressure, mean ultrasound time, degree of nuclear hardness, and amount of postoperative intraocular inflammation between groups. CONCLUSION: Uneventful phacoemulsification involving meticulous cortical and viscoelastic removal does not cause a significant increase in intraocular pressure 24 hours following surgery. PMID- 16355951 TI - Preliminary results of combined simultaneous transpupillary thermotherapy and ICG based photodynamic therapy for choroidal melanoma. AB - BACKGROUND AND OBJECTIVE: To evaluate whether the combination of simultaneous hyperthermia by transpupillary thermotherapy and indocyanine green-based photodynamic therapy is an effective treatment for small and medium choroidal melanomas. PATIENTS AND METHODS: Twenty-five patients with small and medium choroidal melanomas were treated with combined simultaneous transpupillary thermotherapy and indocyanine green-based photodynamic therapy. RESULTS: The median age of the 25 patients was 64 years (range, 35 to 88 years). The pretreatment volume of the tumors ranged from 15.9 to 653.5 mm3 (mean, 118.7 +/- 146.6 mm3). After a mean of 2.4 treatments (range, 1 to 5 treatments), all of the tumors but one showed a significant volume reduction without clinical evidence of recurrences. The follow-up ranged from 6 to 59 months (mean, 12 +/- 14 months). Complications included retinal vascular occlusions, edema and superficial scarring of the macula, and rhegmatogenous retinal detachment. CONCLUSIONS: The effects of combined simultaneous transpupillary thermotherapy and indocyanine green-based photodynamic therapy appears to be effective in achieving local tumor control in selected small and medium choroidal melanomas. PMID- 16355952 TI - Optical coherence tomography in group 2A idiopathic juxtafoveolar telangiectasis. AB - BACKGROUND AND OBJECTIVE: To describe the optical coherence tomographic features in patients with group 2A idiopathic juxtafoveolar telangiectasis. PATIENTS AND METHODS: Forty eyes of 20 consecutive patients with idiopathic juxtafoveolar telangiectasis seen between August 2002 and January 2004 were included in the study. All of the patients underwent color fundus photography, fluorescein angiography, and optical coherence tomography. The main outcome measure was optical coherence tomography findings. RESULTS: The most consistent finding between stages 2 through 5 of group 2A idiopathic juxtafoveolar telangiectasis seen in 35 (87.5%) eyes on optical coherence tomography was the presence of hyporeflective intraretinal spaces in the absence of retinal thickening. Other findings included the presence of hyperreflectivity in the middle or inner retinal layers, suggesting retinal pigment epithelium proliferation and migration corresponding to the stellate foci of pigmentation in stage 4 and features of choroidal or subretinal neovascular membrane in stage 5. CONCLUSION: The optical coherence tomography findings in group 2A idiopathic juxtafoveolar telangiectasis were characteristic and may be helpful in making the diagnosis and defining, as far as possible, the anatomical staging. PMID- 16355953 TI - Extraocular muscle imaging in complex strabismus. AB - BACKGROUND AND OBJECTIVE: Abnormalities of extraocular muscle are an unusual cause of complex strabismus. The traditional evaluation based on clinical examinations is insufficient for the interpretation of incomitant motility disorders resulting from extraocular muscle anomalies. Extraocular muscle imaging by computed x-ray tomography (CT) or magnetic resonance imaging (MRI) can provide useful information for diagnosis, pathophysiology, and treatment of complex strabismus. PATIENTS AND METHODS: Five cases of complex strabismus resulting from congenital anomalies of the extraocular muscles and their successful evaluation using extraocular muscle imaging are described. RESULTS: Orbital CT or MRI scan was obtained in five patients who had unusual incomitant strabismus. It confirmed the diagnosis of the absence of the medial rectus muscle, accessory lateral rectus muscle, atrophy of the inferior or both superior and medial rectus muscles, and abnormal thickening of the levator palpebrae superioris and superior rectus muscle. CONCLUSIONS: Extraocular muscle imaging is a useful technique for evaluating anatomic abnormalities. It should be considered when evaluating patients with atypical strabismus. PMID- 16355954 TI - Retinal nerve fiber analysis and tomography of the optic disc in eyes with tilted disc syndrome. AB - BACKGROUND AND OBJECTIVE: To investigate retinal nerve fiber layer thickness and optic disc parameters in eyes with tilted disc syndrome. PATIENTS AND METHODS: Thirty-eight eyes with tilted disc syndrome (20 myopic control eyes, and 20 emmetropic control eyes) were examined. After a routine ophthalmologic examination of all eyes, scanning laser polarimetry (for retinal nerve fiber layer thickness) and scanning laser tomography (for optic disc parameters) were performed. Scanning laser polarimetry and scanning laser tomography parameters were compared in the different groups. RESULTS: On scanning laser polarimetry evaluation, statistically significant differences were found in the superior integral, nasal integral, and superior nasal quadrants of eyes with tilted disc syndrome compared with control groups (P = .01, P = .04, P = .00, P = .00, P = .00, and P = .00, respectively). On scanning laser tomography evaluation, statistically significant differences were found in average diameter, total contour area, effective area, average depth, volume above, neuroretinal rim area, and cup-disc ratio in eyes with tilted disc syndrome compared with control groups (P = .00, P = .00, P = .00, P = .04, P = .02, P = .00, and P = .01, respectively). CONCLUSION: Parameters acquired through imaging with scanning laser polarimetry and scanning laser tomography in eyes with tilted disc syndrome are different from those of normal eyes. PMID- 16355955 TI - An anterior chamber mass discovered after routine cataract surgery. AB - A 68-year-old man was noted to have an anterior chamber mass shortly after uneventful cataract surgery. The mass was believed to be iatrogenic in nature and necessitated surgical removal because of intermittent obstruction of vision. The patient was observed closely and achieved a good visual outcome with no complaints. This case is described to illustrate a potential hazard of modern clear cornea cataract surgery. PMID- 16355957 TI - Chorioretinal neovascularization after radial optic neurotomy for central retinal vein occlusion. AB - Radial optic neurotomy was recently introduced for the treatment of central retinal vein occlusion. Two patients developed chorioretinal neovascularization through the radial cut of the optic disc after pars plana vitrectomy, radial optic neurotomy, and endophotocoagulation. Patients undergoing radial optic neurotomy should be closely observed to minimize the risk of this complication. PMID- 16355956 TI - Delayed-onset endophthalmitis following cataract surgery caused by Acremonium strictum. AB - This case represents the first report of Acremonium strictum endophthalmitis. Endophthalmitis caused by A. strictum may share clinical features typical of endophthalmitis due to Propionibacterium acnes (such as delayed onset of the infection and white plaque-like material in the capsular bag). PMID- 16355958 TI - Phacoemulsification and lens-iris diaphragm retropulsion syndrome. AB - Lens-iris diaphragm retropulsion syndrome is mainly caused by a reverse pupillary block as a result of a defect or laxity in the zonular apparatus. It is characterized by a posterior displacement of the lens-iris diaphragm with posterior iris bowing, pupil dilation, and pain. The authors describe a new management technique using an iris hook retractor to lift the iris to relieve the pupillary block. PMID- 16355959 TI - Treatment of retinal breaks with large-spot diode laser photocoagulation. AB - A new technique is described for treating peripheral retinal breaks using large spot, diode laser retinopexy. A large-spot (1.2 to 3.0 mm) diode laser (810 nm) was used to treat patients with breaks in attached retina. Powers between 800 and 1,200 mW produced a uniform retinal photocoagulation beyond the break margins. Patients were observed monthly until an adequate chorioretinal scar had formed. Eleven eyes were treated using this technique. Satisfactory retinopexy was obtained in all cases. The efficiency of large-spot diode laser photocoagulation for retinal breaks and its advantages over conventional laser and cryopexy techniques justifies further studies into this technique. PMID- 16355960 TI - Sutureless limited vitrectomy for positive vitreous pressure in cataract surgery. AB - A sutureless transconjunctival pars plana vitrectomy with the 25-gauge transconjunctival vitrectomy system is used to facilitate phacoemulsification in eyes with positive posterior vitreous pressure and shallow anterior chamber. Peribulbar local anesthesia is administered. In eyes with shallow anterior chamber, if an injection of a viscoelastic substance through anterior chamber paracentesis fails to deepen the anterior chamber, a limited pars plana vitrectomy is performed to remove a small amount of retro-lental vitreous (approximately 0.2 to 0.3 cc) with a 25-gauge high-speed cutter. Phacoemulsification is subsequently performed. The limited pars plana vitrectomy reduces the chances of intraoperative vitreous loss and suprachoroidal hemorrhage. It also increases the anterior chamber depth, facilitates intraoperative steps such as pupil stretching and capsulorhexis, and results in a phacoemulsification procedure that is less complex and safer. PMID- 16355961 TI - An improved water bath for ultrasound biomicroscopy. AB - A beveled, oval-shaped eyecup molded from a medical-grade polymer with a round fluid reservoir fused to the top of it has been designed for use with the ultrasound biomicroscope. This new design addresses the problem of saline leaking out from under the water bath, improves comfort for patients with various lid fissures, and increases room for oscillations of the ultrasound probe. PMID- 16355962 TI - Phases in titanium alloys. AB - Titanium (Ti) provides the basis for many alloys used in medical technology. These alloys have been discussed many times in this column over the past decade. In recent years in particular, there have been a number of developments in Ti metallurgy, which suggests that a reappraisal of these alloy systems is timely. PMID- 16355963 TI - Selecting silicone tubing for device applications. AB - A number of factors are involved in selecting the most suitable silicone tubing for a given purpose. These include physical chemistry, performance properties, a supplier's quality system and regulatory compliance. This article provides a guide for device developers when selecting silicone tubing for their applications. PMID- 16355964 TI - Material selection: the moulder's view. AB - Where is the best place to turn for help with plastics material selection? Sources of data and expertise are outlined here. PMID- 16355965 TI - Angioplasty catheters: manufacturing and material problems addressed. AB - This article describes a project to determine the causes of a high failure rate in one manufacturer's percutaneous transluminal coronary angioplasty catheters. Recommendations for optimal production are outlined. PMID- 16355966 TI - Conducting clinical studies in Italy. AB - Medical device manufacturers conduct clinical studies in Italy for various reasons, including the size of the Italian market and the presence of leading health-technology centres. This article discusses the manner in which the European requirements for medical device clinical investigations are interpreted and enforced in Italy. PMID- 16355967 TI - Stem cells are big news, but are they big business? AB - Future health care will involve treatment with stem cells and tissue engineering. This article, the first in a series reporting on the scientific breakthroughs in this area and relating them to the medical device industry, describes the science. PMID- 16355968 TI - Circuit board design for good electromagnetic compatibility performance. AB - Design engineers need to consider the electromagnetic performance of printed circuit boards to adhere to electromagnetic compatibility requirements. This article examines factors that influence performance and outlines techniques to avoid the use of overly expensive screen enclosures in order to comply with the standards. PMID- 16355969 TI - Insights on ideation and innovation. AB - Even when companies are struggling or seeking to cut costs, it is still important for them to generate new ideas. This article discusses the concept of ideation and provides strategies for successful innovation. PMID- 16355970 TI - Packaging issues: avoiding delamination. AB - Manufacturers can minimise delamination occurrence by applying the appropriate packaging design and process features. The end user can minimise the impact of fibre tear and reduce subsequent delamination by careful package opening. The occasional inconvenient delamination is a small price to pay for the high level of sterility assurance that comes with the use of Tyvek. PMID- 16355971 TI - A dynamic industry where innovation is a priority. AB - Each year, Medical Device Technology surveys its readers to identify some of the trends and characteristics of the European market. In this, the first of a three part report on the findings, the focus is on innovation and change. PMID- 16355972 TI - Testing the materials used in a needle-free injector. AB - A universal material tester has proved invaluable during the development of one company's needle-free injection system. PMID- 16355973 TI - A novel valve offers designers new opportunities. AB - A miniature valve has been developed that reportedly offers flow rates of more than 200 L/min, depending on supply pressure. The thinking behind the design is reported here. PMID- 16355974 TI - Induction of cysteine dioxygenase activity by oral administration of cysteine analogues to the rat: implications for drug efficacy and safety. AB - One of the major steps in the oxidation of the sulphur-containing amino acid, L cysteine, is the production of cysteine sulphinic acid, catalysed by the enzyme cysteine dioxygenase. This enzyme plays a key role in the intermediary metabolism of sulphur-containing compounds. The activity of this crucial enzyme is known to be influenced by sulphur-compound intake, being increased in animals fed an excess of L-cysteine or methionine. However, the affects on this enzyme of the chronic administration of drugs similar in structure to cysteine are unknown. This has now been investigated using the anti-rheumatic agent, D-penicillamine, and the mucoactive compound, S-carboxymethyl-L-cysteine. Repeated oral administration of these sulphur-containing drugs to male Wistar rats for five consecutive days led to a significant increase in hepatic cysteine dioxygenase activity. This increase in the production rate of cysteine sulphinic acid remained evident until returning to control levels four days after cessation of drug administration. These observations provide evidence that these two drugs interact with the intermediary biochemistry of sulphur compounds and may provide hitherto unappreciated insights into mechanisms by which therapeutic effects and adverse reactions may occur. PMID- 16355975 TI - Interaction between thiopental and fentanyl in mice is different between hypnotic and lethal doses. AB - Much is known about the interaction of intravenous anesthetics and opioids at the therapeutic level, but less is known regarding their combined lethal effect, leaving some uncertainty regarding the window of safety for their clinical use. We set out to document the type of interaction between thiopental and fentanyl for both the hypnotic effect (loss of righting reflex) and lethal effect in mice. Hypnotic and lethal dose-response curves were constructed for thiopenthal alone and in combination with fentanyl (0.8 microg/kg, each based on five to seven subgroups of six to ten ICR mice. The dose of fentanyl was that needed to double the lag time to tail flick following a noxious stimulus (the equivalent of human analgesia). While fentanyl did not change the median effective hypnotic dose of thiopental (8.9 mg/kg [95% confidence interval {CI} 8.0-9.9 mg/kg] alone versus 7.8 mg/kg [95% CI 6.7-8.7 mg/kg] in combination), it significantly reduced its median lethal dose from 71.8 mg/kg (95% CI 68.3-74.8 mg/kg) to 64.5 mg/kg (95% CI 63.7-65.2 mg/kg). Most remarkably, it increased the slope of the curve from 0.17 (95% CI 0.10-0.36) to 0.61 (95% CI 0.24-1.10), virtually eliminating the difference between the non-lethal and lethal ranges. We conclude that the type of interaction between thiopental and fentanyl is stronger for the lethal effect than for the hypnotic effect. This may become relevant to clinical situations in humans when higher doses of thiopental are used. PMID- 16355976 TI - The effects of vigabatrin on rat liver antioxidant status. AB - The anti-epileptic drug vigabatrin was developed as an inhibitor of gamma aminobutyric acid transaminase, and its ability to increase inhibition in the central nervous system led to its testing in an animal model. In animal models chronic use of vigabatrin is associated with irreversible myelin vacuolation. Antioxidant drugs change the antioxidant capacity of the body. Oxidative stress of the body increased when valproic acid and carbamazepine were used chronically. To assess whether vigabatrin may affect protein oxidation and lipid peroxidation, glutathione, glutathione peroxidase (GPx), and glutathione-S-transferase (GST) levels were studied in the livers of 57 rat fetuses after administration of vigabatrin to the mothers (19 in the first week of pregnancy, 20 in the second week, and 18 in the third week) and in 19 control rat fetuses without vigabatrin. We compared the results of administration of vigabatrin in each group with the controls. Rat fetus protein oxidation in group I (0.686 nmol/mg protein) and group II (0.723 nmol/mg protein) was higher than in the control group (0.388 nmol/mg protein). Lipid peroxidation (0.209, 0.224, 0.253 nmol/mg protein, respectively) and GPx levels (345.4, 329.0, 283.2 nmol/mg protein, respectively) of groups I, II, and III were higher than in the control group (0.104, 167.2 nmol/mg protein, respectively). GST in group II (79.2 nmol/mg protein) and group III (77.8 nmol/mg protein) were not different from that in the control group (78 nmol/mg protein). It was found that vigabatrin affected all the parameters that were studied, especially in group I, which was given the drug in the first week of pregnancy. PMID- 16355977 TI - Interactions between ciprofloxacin and antacids--dissolution and adsorption studies. AB - Ciprofloxacin is a fluorinated quinolone antibacterial agent extensively used against both Gram-positive and Gram-negative microorganisms. In certain polytherapy programs, ciprofloxacin can be administered with some antacids that could modify its dissolution rate and reduce its absorption leading to therapeutic failure. The aim of this study was to evaluate the influence of some antacids on the availability of ciprofloxacin. The release of ciprofloxacin from tablets in the presence of antacids, such as sodium bicarbonate, calcium hydroxide, calcium carbonate, aluminum hydroxide, magnesium hydroxide, magnesium carbonate, magnesium trisilicate and magaldrate was studied on BP 2002 dissolution test apparatus. These studies were carried out in simulated gastric and intestinal juices for 3 hours at 37 degrees C. The results confirmed that the dissolution rate of tablets was markedly retarded in the presence of all the antacids studied. Magaldrate and calcium carbonate in simulated gastric juice exhibited relatively higher adsorption capacities, as did magnesium trisilicate and calcium hydroxide in simulated intestinal juice. PMID- 16355978 TI - Determination of the antimicrobial properties of oligo-2-hydroxy-1 naphthaldehyde. AB - Oligo-2-hydroxy-1-naphthaldehyde (OHNA) was synthesized by oxidative polycondensation using H2O2 (35%, aqueous solution), air O2 and NaOCl (34%, aqueous solution) by Kaya and Senol and the products were characterized by spectral techniques. Antimicrobial activities of the first and second fractions of OHNA were tested against Corynobacterium xerosis CCM 2824, Proteus vulgaris ATCC 6897, Staphylococcus epidermidis NRRL B-4877, S. aureus ATCC 6538, Enterobacter aerogenes ATCC 13048, Salmonella thyphimurium CCM 5445, Pseudomonas aeroginosa ATCC 27853, Escherichia coli ATCC 11230, E. coli ATCC 23998, Bacillus cereus ATCC 7064, B. cereus ATCC 99, B. subtilis ATCC 6633, Yersinia spp., Neisseria canis, Rhodotorula rubra, Kluyveromyces fragilis NRRL 2415, Saccharomyces cerevisiae ATCC 9763, S. ovarum, Debaryomyces hensenii, Hansenula anamola, Candida albicans, C. utilis, Aspergillus niger, A. fumigates, A. versicolor, A. flavus, A. parasiticus, Penicillium granulatum, P. chrysogenum, and P. herque. OHNA demonstrated antimicrobial activity against various bacteria and yeast, but did not affect filamentous fungi. PMID- 16355979 TI - Total synthesis of guanacastepene a: a route to enantiomeric control. AB - [reaction: see text] The goal of the total synthesis of guanacastepene A served as a focus to bring together several chemical inquiries. One involved the synthesis of fused 5,7-hydrazulenones (see structure 20). Another issue had to do with the mechanistic intermediates in reductive cyclizations (see 17 to 18 and 19). The total synthesis required a mastery of an intramolecular Knoevenagel condensation of a beta,gamma-unsaturated ketone (see compound 41). Actually, cyclization was best accomplished when the terminal double bond of 41 was first converted to an epoxide. Further issues related to the stereochemistry at C5 and, rather surprisingly, the propensity for beta-face acetoxylation at C13. Crystallographic verification of the assigned beta-stereochemistry at C13 is provided. Finally, a route to optically active material is provided (see compound 20). A key element in this construction was an enantioselective addition of isopropenyl cuprate to 2-methylcyclopentenone (see compound 99). PMID- 16355980 TI - Is the Beckmann rearrangement a concerted or stepwise reaction? A computational study. AB - [reaction: see text] RB3LYP calculations were performed on the Beckman rearrangement by the use of three substrates, acetone oxime (1), acetophenone oxime (2), and cyclohexanone oxime (3). Acidic solvents were modeled by H+ (CH3COOH)3 and H3O+ (H2O)6, and reaction paths were determined precisely. For 1, a two-step process involving a sigma-type cationic complex was obtained. For 2, a three-step process with pi- and sigma-type complexes was found in H+ (CH3COOH)3 and a two-step process involving a sigma-type cationic complex was obtained in H3O+ (H2O)6. However, for 3, a concerted process without pi and sigma complexes was calculated, which leads to the product, epsilon-caprolactam. Three different mechanisms were explained in terms of FMO theory. PMID- 16355982 TI - Pentafluorophenyl transfer: a new group-transfer reaction in organoborate salts. AB - [reaction: see text] Irradiation of isoquinolinium hydroxytris(pentafluorophenyl)borate, 1, and phenanthridium hydroxytris(pentafluorophenyl)borate, 2, in either CH2Cl2 or CH3CN resulted in C6F5 transfer to the isoquinolinium and phenanthridium cations, generating 2 methyl-1-(2,3,4,5,6-pentafluorophenyl)-1,2-dihydroisoquinoline, 3, and 2-methyl-1 (2,3,4,5,6-pentafluorophenyl)-1,2-dihydrophenanthridine, 4, respectively. In addition, photogeneration of H2O x B(C6F5)3 resulted from 1. Photogeneration of C6F5-C6F4H from HO-B(C6F5)3(-) and of C6H5-C6F4H from C6H5-B(C6F5)3(-) was discovered. PMID- 16355981 TI - Revisiting a classic approach to the Aspidosperma alkaloids: an intramolecular Schmidt reaction mediated synthesis of (+)-aspidospermidine. AB - [reaction: see text] A total synthesis of (+)-aspidospermidine (1) is described. The key reactions used in the synthesis of this pentacyclic Aspidosperma alkaloid were a deracemizing imine alkylation/Robinson annulation sequence, a selective "redox ketalization", and an intramolecular Schmidt reaction. A Fischer indolization step carried out on a tricyclic ketone mirrored the sequence reported by Stork and Dolfini in their classic aspidospermine synthesis. PMID- 16355983 TI - Synthesis of crescent aromatic oligoamides. AB - [structures: see text] This article describes the synthetic procedures for the preparation of crescent (and helical) aromatic oligoamides developed in recent years in our laboratory. The large-scale preparation of a variety of monomers derived from various tetrasubstituted benzenes is presented. Three different strategies for constructing various oligomers consisting of meta- and meta/para linked benzene residues are discussed. Factors affecting coupling efficiency and yields are analyzed. The developed synthetic methods have provided the basis for the preparation of longer oligomers and for the development of solid-phase synthesis. PMID- 16355985 TI - Rare-earth silylamide-catalyzed monocoupling reaction of isocyanides with terminal alkynes. AB - [reaction: see text] Rare-earth silylamides, Ln[N(SiMe3)2]3 (Ln = Y, La, Sm, Yb), serve as good catalysts for monoinsertion of isocyanides into terminal alkynes in the presence of amine additives, leading to 1-aza-1,3-enyens in excellent yields. The reaction is applicable to a diverse set of terminal alkynes with various functionalities such as ethers, acetals, and amino groups. Larger metals (La and Sm) give a better performance than smaller ones (Y and Yb). Using less hindered primary amines and, in contrast, bulky isocyanides is crucial for the coupling reaction; otherwise, competitive oligomerization of the isocyanides occurs predominantly. In the mechanistic study, the rate-determining step of the reaction seems to be the first insertion of the isocyanides into rare-earth alkynides, which is followed by spontaneous protonation with the amine additives. PMID- 16355984 TI - Propagation of polar substituent effects in 1-(substituted phenyl)-6,7-dimethoxy 3,4-dihydro- and -1,2,3,4-tetrahydroisoquinolines as explained by resonance polarization concept. AB - [structures: see text] Propagation of inductive and resonance effects of phenyl substituents within 1-(substituted phenyl)-6,7-dimethoxy-3,4-dihydro- and 1,2,3,4-tetrahydroisoquinolines were studied with the aid of 13C and 15N NMR chemical shifts and ab initio calculations. The substituent-induced changes in the chemical shift (SCS) were correlated with a dual substituent parameter equation. The contributions of conjugative (rhoR) and nonconjugative effects (rhoF) were analyzed, and mapping of the substituent-induced changes is given over the entire isoquinoline moiety for both series. The experimental results can be rationalized with the aid of the resonance polarization concept. This means the consideration of the substituent-sensitive balance of different resonance structures, i.e., electron delocalization, and the effect of the aromatic ring substituents on their relative contributions. With tetrahydroisoquinolines, the delocalization of the nitrogen lone pair (stereoelectronic effect) particularly contributes. Correlation analysis of the Mulliken atomic charges for the dihydroisoquinoline derivatives was also performed. The results support the concept of the substituent-sensitive polarization of the isoquinoline moiety even if the polarization pattern achieved via the NMR approach is not quite the same as that predicted by the computational charges. Previously the concepts of localized pi-polarization and extended polarization have been used to explain polar substituent effects within aromatic side-chain derivatives. We consider that the resonance polarization model effectively contributes to the understanding of the polar substituent effects. PMID- 16355986 TI - Structure and reactivity of trans-bis[2-(2-chloroethyl)pyridine]palladium chloride (1). A study on the elimination reaction of 1 and 2-(2 chloroethyl)pyridine induced by quinuclidine in acetonitrile. AB - [reaction: see text] The trans-bis[2-(2-chloroethyl)pyridine]palladium chloride (1) has been prepared and structurally characterized by X-ray spectroscopy and computational study. The X-ray structure of 1 is consistent with the trans isomer (with respect to Pd). The NMR spectrum and the computational study are in agreement with an equilibrium in CD3CN solution between two isomers of the trans structure. The reaction of the palladium complex with quinuclidine in CH3CN, at 25 degrees C, leads to competing elimination and displacement reactions with formation of vinylpyridine and chloroethylpyridine in a ratio of 1.5:1. However, the rate constant for formation of uncoordinated (vinyl)pyridine monitored by HPLC (kQ(HPLC) = 2.3 x 10(-3) M(-1) s(-1)) is nearly 3 times slower than a rate constant monitored spectrophotometrically (kQ = 6.5 x 10(-3) M(-1) s(-1)). This suggests that the initial product of elimination is a palladium complex of vinylpyridine and that displacement from this complex is partially rate determining in the formation of the uncoordinated product. A study by UV spectroscopy at lambda = 295 nm of trans-bis[2-(2-chloroethyl)pyridine d2]palladium chloride with quinuclidine (Q) has shown the presence of a significant primary kinetic isotope effect, kQ(H)/kQ(D) = 1.8, for the elimination reaction within the Pd complex, 1. The second-order rate constant for the beta-elimination reaction from 2-(2-chloroethyl)pyridine induced by quinuclidine in CH3CN at 25 degrees C is kQ(FREE) = 6.2 x 10(-6) M(-1) s(-1). It can be observed as a significant activation (about 3 orders of magnitude) of the beta-elimination reaction within the complex 1 with respect to the free 2-(2 chloroethyl)pyridine. The possible mechanism in agreement with these results is discussed. PMID- 16355987 TI - Reductive cleavage of tetrahydrofuryl sulfur-substituted oxiranes: application to the formal synthesis of Kumausyne and Kumausallene. AB - [reactions: see text] Readily available sulfinyl and sulfonyl tetrahydrofuran methanol derivatives have been transformed efficiently into a variety of substituted tetrahydrofuryl alcohols by treatment with (PhSe)2 in the presence of an excess of NaBH4. Alternatively, oxirane cleavage with MgI2 produces the related ketones, amenable to stereocontrolled reduction. This reductive cleavage methodology has been applied to short formal syntheses of trans-Kumausyne and Kumausallene. PMID- 16355988 TI - Solid-phase synthesis of "mixed" peptidomimetics using Fmoc-protected aza-beta3 amino acids and alpha-amino acids. AB - [structure: see text] A solid-phase fluorenylmethyloxycarbonyl (Fmoc)-based synthesis strategy is described for "mixed" aza-beta3-peptides as well as a convenient general approach for their required building blocks, the aza-beta3 amino acid residues (aza-beta3-aa). These monomers allow the synthesis of relatively large quantities of pure mixed aza-beta3-peptides. The required Fmoc substituted aza-beta3-amino acids are accessible by convenient synthesis, and a number of monomers including those containing side chains with functional groups have been synthesized. The method was applied toward the solid-phase synthesis of aza-beta3-peptide mimetics of a biologically active histone H4 sequence. PMID- 16355989 TI - Dications of fluorenylidenes. The relationship between redox potentials and antiaromaticity for meta- and para-substituted diphenylmethylidenefluorenes. AB - [reaction: see text] Electrochemical oxidation of meta-substituted diphenylmethylidenefluorenes (3a-g) results in the formation of fluorenylidene dications that are shown to be antiaromatic through calculation of the nucleus independent chemical shift (NICS) for the 5- and 6-membered rings of the fluorenyl system. There is a strong linear correlation between the redox potential for the dication and both the calculated NICS and sigma(m). Redox potentials for formation of dications of analogously substituted tetraphenylethylenes shows that, with the exception of the p-methyl derivative, the redox potentials for these dications are less positive than for formation of the dications of 3a-g and for dications of p-substituted diphenylmethylidenefluorenes, 2a-g. The greater instability of dications of 2a-g and 3a-g compared to the reference system implies their antiaromaticity, which is supported by the positive NICS values. The redox potentials for formation of the dications of meta-substituted diphenylmethylidenes (3a-g) are more positive than for the formation of dications of para-substituted diphenylmethylidenes (2a-g), indicating their greater thermodynamic instability. The NICS values for dications of 3a-g are more antiaromatic than for dications of 2a-g, which is consistent with their greater instability of the dications of 3a-g. Although the substituted diphenylmethyl systems are not able to interact with the fluorenyl system through resonance because of their geometry, they are able to moderate the antiaromaticity of the fluorenyl cationic system. Two models have been suggested for this interaction, sigma to p donation and the ability of the charge on the substituted ring system to affect delocalization. Examination of bond lengths shows very limited variation, which argues against sigma to p donation in these systems. A strong correlation between NICS and sigma constants suggests that factors that affect the magnitude of the charge on the benzylic (alpha) carbon of the diphenylmethyl cation affect the antiaromaticity of the fluorenyl cation. Calculated atomic charges on carbons 1-8 and 10-13 show an increase in positive charge, and therefore greater delocalization of charge in the fluorenyl system, with increasing electronegativity of the substituent. The change in the amount of positive charge correlated strongly with NICS, supporting the model in which the amount of delocalization of charge is related to the antiaromaticity of the species. Thus, both aromatic and antiaromatic species are characterized by extensive delocalization of electron density. PMID- 16355990 TI - Hydrosilylation of terminal alkynes with alkylidene ruthenium complexes and silanes. AB - [reaction: see text] Ruthenium alkylidene complexes 1-3 mediate hydrosilylation of alkynes with silanes. When triethoxy- or triphenylsilanes are used as silylating agents, the reaction affords alpha-substituted vinylsilanes as major products. PMID- 16355991 TI - Intramolecular cycloaddition of O-tert-butyldimethylsilyloximes in the presence of BF3 x OEt2. AB - [reaction: see text] Intramolecular cycloaddition of novel 1,3-dipoles, N boranonitrones, was examined. Treatment of O-tert-butyldimethylsilyloximes 9-12 having olefin moieties with 2 equiv of BF3 x OEt2 generated N-boranonitrones, which underwent intramolecular cycloaddition to afford N-nonsubstituted cycloadducts 16 (and/or 18) after extractive workup. Despite the Lewis-acidic conditions, the olefin geometry of the substrates was retained in the cycloadducts in the present cycloaddition. The electronic nature of the N boranonitrones appeared to be electrophilic. In the case of substrate 11c, having an electron-donating methyl group at an internal position of the olefin moiety, the cycloaddition gave the bridged cycloadduct 18b. The cycloaddition proceeded at relatively low temperature, and the diastereoselectivity was high. PMID- 16355992 TI - Conformational study of cis-1,4-di-tert-butylcyclohexane by dynamic NMR spectroscopy and computational methods. Observation of chair and twist-boat conformations. AB - [reaction: see text] Low-temperature 13C NMR spectra of cis-1,4-di-tert butylcyclohexane (1) showed signals for the twist-boat (1a) and chair (1b) conformations. 13C NMR signals were assigned to specific carbons based on the different populations, different symmetries (time-averaged C(2v) for 1a and time averaged C(s) for 1b), and calculated chemical shifts (GIAO, HF/6-311+G*). In addition to slow ring inversion and interconversion of the chair and twist-boat conformations, slow rotation of the tert-butyl groups was found. Most of the expected 13C peaks were observed. Free-energy barriers of 6.83 and 6.35 kcal/mol were found for interconversion of 1a (major) and 1b (minor) at -148.1 degrees C. Conformational space was searched with Allinger's MM3 and MM4 programs, and free energies were obtained for several low-energy conformations 1a-c. Calculations were repeated with ab initio methods up to the HF/6-311+G* level. Molecular symmetries, relative free energies, relative enthalpies and entropies, frequencies, and NMR chemical shifts were obtained. A boat conformation (1d; C(2v) symmetry) was generated and optimized as a transition state by ab initio, MM3, and MM4 calculations. PMID- 16355994 TI - Asymmetric intermolecular C-H functionalization of benzyl silyl ethers mediated by chiral auxiliary-based aryldiazoacetates and chiral dirhodium catalysts. AB - [reaction: see text] C-H functionalization of benzyl silyl ethers by means of rhodium-catalyzed insertions of aryldiazoacetates can be achieved in a highly diastereoselective and enantioselective manner by judicious choice of chiral catalyst or auxiliary. The dirhodium tetraprolinates such as Rh2((S)-DOSP)4 have been widely successful as chiral catalysts in the C-H functionalization chemistry of aryldiazoacetates, but give poor enantioselectivity in the reactions of aryldiazoacetates with benzyl silyl ether derivatives. The use of (S)-lactate as a chiral auxiliary resulted in C-H functionalization with moderately high diastereoselectivity (79-88% de) and enantioselectivity (68-85% ee). The best results (91-95% de, 95-98% ee), however, were achieved using Hashimoto's Rh2((S) PTTL)4 catalyst. PMID- 16355993 TI - Synthesis of the shark repellent pavoninin-4. AB - [reaction: see text] The first synthesis of the shark repellent pavoninin-4, 3, was achieved in 12 steps with 21% overall yield from diosgenin, 8. Key reactions involve an efficient synthesis of the C-15alpha hydroxyl steroid from a C-16beta hydroxyl steroid by an unexpected 1,2-transposition strategy, a stereospecific glycosylation of a hindered C-15alpha alcohol using glycosyl fluoride as a glycosyl donor and a highly chemoselective acetylation of the C-26 primary alcohol by catalytic transesterification. PMID- 16355995 TI - Stereospecific preparation of (Z)- and (E)-2,3-difluoro-3-stannylacrylic ester synthons and a new, efficient stereospecific route to (Z)- and (E)-2,3 difluoroacrylic esters. AB - [reaction: see text] The (Z)-2,3-difluoro-3-stannylacrylic ester is readily prepared from (Z)-1,2-difluorovinyltriethylsilane via stereospecific stannyl/silyl exchange with KF/(Bu3Sn)2O or Bu3SnCl in DMF at 70 degrees C. The corresponding (E)-2,3-difluoro-3-stannylacrylate is prepared by stereospecific carbonylation of (E)-1,2-difluorovinyl iodide followed by low temperature/in situ stannylation of the resultant (Z)-2,3-difluoroacrylic ester. With Cu(I) iodide and Pd(PPh3)4 catalysis, the (Z)- and (E)-stannylacrylate esters readily couple with aryl iodides and vinyl bromides, as well as 2-iodothiophene, at room temperature to stereospecifically produce the respective (E)- and (Z)-2,3 difluoro-3-aryl substituted acrylic esters or conjugated dienes in high yields. PMID- 16355996 TI - Intramolecular ene reaction of epoxyallylsilanes: synthesis of allyl- and vinylsilane-functionalized cyclohexanols. AB - [reaction: see text] Epoxyallylsilanes bearing the bulky tert-butyldiphenylsilyl group undergo an uncommon tandem rearrangement-cyclization process upon treatment with Lewis acids. Two pathways for the carbonyl ene reaction are observed: one leading to allylsilane-cyclohexanols when the epoxyallylsilane (28-31) is nonsubstituted, 2-, or 4-monosubstituted and other leading to vinylsilane cyclohexanols when the epoxyallylsilane (24-27) is 2,4-disubstituted or trisubstituted. An explanation for the observed regio- and stereoselectivity is advanced and a reliable mechanism proposed. PMID- 16355997 TI - Structural investigation of "cis" and "trans" vinylogous peptides: cis-vinylog turn in folded cis-vinylogous peptides, an excellent mimic of the natural beta turn. AB - [structure: see text] Various sequences of modified peptides including those containing a cis- or trans-vinylogous residue have been studied using X-ray diffraction in the solid state and 1H NMR and IR spectroscopy in solution. A cis vinylogous residue promotes an NH to CO intramolecular H-bond, closing a nine membered pseudocycle that stabilizes a folded moiety that we proposed to name the cis-vinylogous turn. A trans-vinylogous residue involves an extended conformation. Two consecutive vinylogous residues retain their own structural propensity: "Xaa(tr)"-"Xaa(cis)" or "Xaa(cis)"-"Xaa(tr)" sequence is singly folded, whereas "Xaa(cis)"-"Xaa(cis)" sequence is doubly folded. Oligo vinylogs with all-trans or all-cis or alternating cis-trans motifs could constitute new classes of foldamers. PMID- 16355998 TI - RCM of tripeptide dienes containing a chiral vinylcyclopropane moiety: impact of different Ru-based catalysts on the stereochemical integrity of the macrocyclic products. AB - [structures: see text] Tripeptide dienes containing an (1R,2S)-vinyl aminocyclopropylcarboxylate residue were cyclized to beta-strand scaffolds under ring-closing metathesis (RCM). Conformational factors, ligand effects, and reaction conditions were evaluated. A protocol was developed for the efficient synthesis of 15-membered ring peptides in high diastereomeric purity. These peptides are key synthetic precursors to antiviral agents that target the hepatitis C virus and represent the first class of clinically validated pharmaceutical agents that are synthesized in large scale using RCM. PMID- 16355999 TI - Synthesis of symmetrical organic carbonates via significantly enhanced alkylation of metal carbonates with alkyl halides/sulfonates in ionic liquid. AB - [reaction: see text] We report a new phosgene-free method for the synthesis of symmetrical organic carbonates via alkylation of metal carbonate with various alkyl halides and sulfonates in 1-n-butyl-3-methylimidazolium hexafluorophosphate, [bmim][PF6], as an ecofriendly reaction media. Alkylation of metal carbonate in various ionic liquids with 1-bromo-3-phenylpropane (1a) as a model reactant has thoroughly been investigated. Potassium and cesium carbonates appeared to be the most suitable metal carbonate due to their high solubility in ionic liquids. Besides good to excellent yields, this simple and convenient methodology is devoid of highly toxic and harmful chemicals such as phosgene and carbon monoxide, which is an additional advantage. PMID- 16356000 TI - Mixed effect of the supporting electrolyte and the zinc anode in the electrochemical homocoupling of 2-bromopyridines catalyzed by nickel complexes in an undivided cell. AB - [reactions: see text] Nickel-catalyzed electroreductive homocoupling of 2 bromomethylpyridines and 2-bromopyridine has been investigated in an undivided cell in the presence of a zinc sacrificial anode. A series of reactions were performed with various types and concentrations of supporting electrolyte. It was observed that a key step in this process is the formation of an arylzinc through a nickel-zinc transmetalation. This intermediate can be transformed back to the reactive arylnickel species to afford the homocoupling as the final product. The back process from the arylzinc intermediate is, however, suppressed in the presence of high concentration (0.2 M) of tetraalkylammonium salts. On the contrary, with NaI, the formation of the dimer is not prevented, whatever the NaI concentration. PMID- 16356001 TI - Efficient catalytic effects of Lewis acids in the 1,3-dipolar cycloaddition reactions of carbonyl ylides with imines. AB - [reactions: see text] 1,3-Dipolar cycloaddition reactions between imines and carbonyl ylides generated by tandem intramolecular carbenoid-carbonyl cyclizations were found to be effectively catalyzed by Lewis acids (10 mol %). The Rh2(OAc)4-catalyzed reactions of o-(methoxycarbonyl)-alpha-diazoacetophenone with imines such as N-[2-(benzyloxy)benzylidene]aniline in the absence of Lewis acid gave no 1,3-dipolar cycloaddition products, but rather the dimeric product of the corresponding carbonyl ylide. In contrast, in the presence of Lewis acids such as Yb(OTf)3, the 1,3-dipolar cycloaddition reactions of the corresponding 1 methoxy-2-benzopyrylium-4-olate proceeded smoothly with several imines, giving in most cases exo-selectivity and no formation of the dimeric product. When Yb(OTf)3 was used as a Lewis acid catalyst, a fundamental catalytic effect was also observed in the cycloaddition reactions of imines with carbonyl ylides generated from 1-diazo-5-phenyl-2,5-pentanedione, 1-diazo-2,5-hexanedione and diazomethyl 2,3,4,5-tetrachloro-6-methoxycarbonylphenly ketone. This efficient catalytic effect can be satisfactorily explained in terms of energetics of the cycloaddition in the absence and the presence of Lewis acid by calculations using the ONIOM (B3LYP/6-31G(d):PM3) method. PMID- 16356003 TI - Diastereoselective ring-closing metathesis: synthesis of P-stereogenic phosphinates from prochiral phosphinic acid derivatives. AB - [reaction: see text] The preparation of phosphorus-containing trienes featuring two diastereotopic vinyl moieties followed by a diastereoselective ring-closing metathesis is described. This methodology allowed for the synthesis of novel highly functionalized P-stereogenic heterocycles featuring both an exo- and an endocyclic double bond. An investigation into the factors influencing the diastereochemical outcome of the ring-closing metathesis is also presented, revealing that the geometry of the double bonds conjugated to phosphorus is important and that 1,3-stereoinduction is superior to 1,4-stereoinduction for these reactions. PMID- 16356004 TI - Synthesis of novel tricyclic pyrimido[4,5-b][1,4]benzothiazepines via Bischler Napieralski-type reactions. AB - [reaction: see text] Novel tricyclic pyrimido[4,5-b][1,4]benzothiazepines were readily prepared from 5-amino-4,6-bis(arylthio)pyrimidines and carboxylic acids via Bischler-Napieralski-type reactions. The 6-aryl sulfide group of the resulting pyrimido[4,5-b][1,4]benzothiazepines could be selectively oxidized to its corresponding sulfoxide, which underwent facile substitution reactions when treated with nucleophiles such as an amine. This synthetic strategy provides an efficient way to access a library of novel heterocyclic compounds that are of interest in drug discovery. PMID- 16356002 TI - Squaric acid-based peptidic inhibitors of matrix metalloprotease-1. AB - [structure: see text] A series of squaric acid-peptide conjugates were synthesized and evaluated as inhibitors of MMP-1. The cyclobut-3-enedione core was substituted at the 3-position with several functional groups, such as N(alkyl)OH, -NHOH, and -OH, that are designed to bind to the zinc atom in the active site of the metalloprotease. The 4-position of the cyclobut-3-enedione was derivatized with mono- or dipeptides that are designed to bind in the S1' and S2' subsites of the enzyme, and position the metal chelating group appropriately in the active site for binding to zinc. Positional scanning revealed that -N(Me)OH provided the highest level of inhibition among the chelating groups that were tested, and Leu-Tle-NHMe was the preferred amino acid sequence. A combination of these groups yielded an inhibitor with an IC50 value of 95 microM. For one inhibitor, conversion of one of the carbonyl groups on the cyclobut-3-enedione core to a thiocarbonyl group resulted in a 18-fold increase in potency, and yielded a compound with an IC50 value of 15 microM. PMID- 16356005 TI - Electrochemically controlled hydrogen bonding. Redox-dependent formation of a 2:1 diarylurea/dinitrobenzene2- complex. AB - [reaction: see text] The electrochemistry of 1,2-dinitrobenzene (1,2-DNB), 1,3 dinitrobenzene (1,3-DNB), and 1,4-dinitrobenzene (1,4-DNB) is strongly affected by the presence of 1,3-diphenylurea. In DMF, the second reduction potential of all three DNBs shifts substantially positive in the presence of the urea, indicating very strong hydrogen bonding to the dianions. With 1,2- and 1,3-DNB, the hydrogen bonding leads to irreversible chemistry, likely due to proton transfer from the urea to the dianions. No such irreversible behavior is observed with 1,4-DNB. Instead, the second reduction shifts into the first reduction, producing a single, reversible, two-electron cyclic voltammetric wave at high urea concentrations. Computer simulations show that the changes in wave shape accompanying this process are well accounted for by the stepwise formation of a 1:1 and 2:1 1,3-diphenylurea/DNB2- complex, with sequential binding constants of approximately 5.5 x 10(4) M(-1) and approximately 4.0 x 10(3) M(-1) in DMF. PMID- 16356006 TI - Lewis base catalyzed aldol additions of chiral trichlorosilyl enolates and silyl enol ethers. AB - [structures: see text] The consequences of double diastereodifferentiation in chiral Lewis base catalyzed aldol additions using chiral enoxysilanes derived from lactate, 3-hydroxyisobutyrate, and 3-hydroxybutyrate have been investigated. Trichlorosilyl enolates derived from the chiral methyl and ethyl ketones were subjected to aldolization in the presence of phosphoramides, and the intrinsic selectivity of these enolates and the external stereoinduction from chiral catalyst were studied. In the reactions with the lactate derived enolate, the strong internal stereoinduction dominated the stereochemical outcome of the aldol addition. For the 3-hydroxyisobutyrate- and 3-hydroxybutyrate derived enolates, the catalyst-controlled diastereoselectivities were observed, and the resident stereogenic centers exerted marginal influence. The corresponding trimethylsilyl enol ethers were employed in SiCl4/bisphosphoramide catalyzed aldol additions, and the effect of double diastereodifferentiation was also investigated. The overall diastereoselection of the process was again controlled by the strong external influence of the catalyst. PMID- 16356007 TI - On the origin of cis/trans stereoselectivity in intramolecular Diels-Alder reactions of substituted pentadienyl acrylates: a comprehensive density functional study. AB - [structures: see text] A gas-phase B3LYP/6-31+G(d) study of substituent effects on the stereochemistry of both intramolecular Diels-Alder (IMDA) reactions of 9-E and 9-Z-substituted pentadienyl acrylates and intermolecular Diels-Alder (DA) reactions between butadiene and monosubstituted alkenes and 3-substituted acrylates is reported and involves the calculation of 230 transition structures. It was found that, although exo ("anti-Alder") addition of monosubstituted ethenes to butadiene is the norm, Alder endo selectivity is more widely predicted for 3-substituted methyl acrylate dienophiles, and this was explained in terms of secondary orbital interactions (SOIs). Whereas cis/trans selectivity for IMDA reactions involving 9-E-substituted pentadienyl acrylates generally follows the normal pattern found for the corresponding intermolecular DA reactions, the 9-Z substituted stereoisomers generally displayed trans selectivity that was much stronger than can be attributed to effects of the isolated substituent. This is strikingly so with unsaturated electron-withdrawing substituents whose endo selectivities, displayed in intermolecular DA reactions, are reversed in the IMDA reactions of pentadienyl acrylates. The origin of this anomalous Z-effect is explained in terms of the twist-mode asynchronicity concept of Brown and Houk. These ideas are used to explain the stereochemical outcomes of IMDA reactions of other triene systems. PMID- 16356008 TI - Radical [n + 1] annulations with sulfur dioxide. AB - [reaction: see text] A new methodology for [n + 1] radical annulation using sulfur dioxide as a geminal radical acceptor/donor is presented. This methodology provides a novel route to the formation of five-, six-, and seven-membered cyclic sulfones utilizing a radical chain mechanism under very mild conditions. PMID- 16356009 TI - Stereocontrolled synthesis of (-)-kainic acid from trans-4-hydroxy-L-proline. AB - [reaction: see text] A highly stereoselective synthesis of (-)-kainic acid has been achieved in 14 steps and >7% overall yield starting from inexpensive trans-4 hydroxy-L-proline. The key steps are diastereoselective enolate alkylation and cuprate substitution reactions. PMID- 16356010 TI - Multicomponent one-pot procedure for the synthesis of free alpha-chiral amines from aldehydes. AB - [reaction: see text] The synthesis of free alpha-chiral amines by a one-pot multicomponent procedure from commercially available starting materials is described. This enantioselective reaction involves a catalytic asymmetric addition of dialkylzinc reagents to N-diphenylphosphinoylimines with use of an air-stable precatalyst complex 1. The alpha-chiral amines are prepared with a one pot procedure from alkyl and aryl aldehydes in good yield (41-90%) and with excellent enantioselectivity (90-97% ee). PMID- 16356011 TI - Synthesis of highly functionalized pyrrolidines via a mild one-pot, three component 1,3-dipolar cycloaddition process. AB - [reaction: see text] A simple and efficient one-pot, three-component synthesis of highly functionalized pyrrolidines via cascade imine --> azomethine ylide --> 1,3 dipolar cycloadditions is reported. Admixing a variety of aldehydes, dimethyl 2 aminomalonate, and electron deficient alkenes in THF leads to the clean production of pyrrolidines in good to excellent yields. The mild reaction conditions enabled the generation of previously inaccessible azomethine ylides from enolizable aldehydes. Endo selectivity was exclusive with N-phenyl maleimide and maleic anhydride. Good chemo-, regio-, and stereoselectivities were observed with methyl acrylate, though catalysis by Ag(I) was necessary with this dipolarophile. PMID- 16356012 TI - An expeditious, high-yielding construction of the food aroma compounds 6-acetyl 1,2,3,4-tetrahydropyridine and 2-acetyl-1-pyrroline. AB - [reaction: see text] The key compound responsible for the aroma of bread, 6 acetyl-1,2,3,4-tetrahydropyridine (1), has been constructed in an efficient three step procedure from 2-piperidone in an overall yield of 56%. Compound 1 was liberated in the final step under basic conditions. A related synthetic route produced 2-acetyl-1-pyrroline (2), the principal component of cooked rice, in 10% overall yield. PMID- 16356013 TI - Colorimetric "naked eye" sensing of anions in aqueous solution. AB - [structure: see text] The synthesis and UV-vis and NMR spectroscopic studies of thiourea-based colorimetric sensors for anions are presented. These sensors can recognize anions through hydrogen binding even in competitive pH-buffered aqueous solutions, giving rise to large color changes that are clearly visible to the naked eye. PMID- 16356014 TI - Baeyer-Villiger oxidation with potassium peroxomonosulfate supported on acidic silica gel. AB - [reaction: see text] Potassium peroxomonosulfate deposited onto silica SiO2 x KHSO5 efficiently reacts with ketones in dichloromethane at room temperature to give the corresponding esters or lactones in quantitative yields. This method avoids hydrolysis of the reaction products. The Baeyer-Villiger reaction is catalyzed by potassium hydrogensulfate present in the supported reagent. PMID- 16356015 TI - A new, mild, and efficient synthesis of 2,2-difluoro-3-hydroxyacids through a selective haloform reaction. AB - [reaction: see text] Long-chain 2,2-difluoro-3-hydroxyacids have been synthesized in a new, straightforward manner by treatment of 4-hydroxy-1,1,1,3,3 pentafluoroalkyl ketones, easily obtained by reaction of pentafluoroenolate 2 with aldehydes and ketones, with base under mild conditions. The reaction sequence is marked by the selective cleavage of the CO-CF3 bond, as well as the absence of products arising from the alternative CO-CF2R bond cleavage. The process represents a convenient approach for the synthesis of 2,2-difluoro-3 hydroxyacids, as it is short, provides good to excellent yields under mild conditions, and uses hexafluoro-2-propanol, a very cheap reagent, as the fluorine source. PMID- 16356016 TI - [5C + 1S] annulation: a facile and efficient synthetic route toward functionalized 2,3-dihydrothiopyran-4-ones. AB - [reaction: see text] A facile and efficient synthetic route toward highly substituted 2,3-dihydrothiopyran-4-ones 2 has been developed via a formal [5C + 1S] annulation of readily available alpha-alkenoyl ketene-(S,S)-acetals 1 with sodium sulfide nonahydrated salt (Na2S x 9H2O) and utilized in the synthesis of 2 (4-chlorophenyl)-6-(morpholin-4-yl)-4H-thiopyran-4-one 5l, an inhibitor of DNA dependent protein kinase (DNA-PK). PMID- 16356022 TI - Iron deficiency and impaired cognition in toddlers: an underestimated and undertreated problem. AB - Iron deficiency in toddlers is associated with impaired cognition and is an underestimated and undertreated problem. The prevalence of iron deficiency anemia (IDA) during the first year of life has been dramatically reduced in developed countries, mainly due to the increase in breastfeeding and the use of iron fortified feeding formulae. However, in US and UK children aged 1-2 years, recent studies have shown prevalence rates of >10% and 30% for IDA and iron deficiency, respectively. The daily iron intake in children aged 1-2 years is lower than in any other age group during life. IDA during the first 2 years of life is associated with impaired mental and psychomotor development and these deficits are long lasting, and perhaps irreversible, despite the correction of the anemia. Another compelling reason to prevent iron deficiency in children, especially in children aged 1-2 years, is the proven association of iron deficiency with increased lead absorption. Lead-associated cognitive deficits occur at blood lead levels <10 microg/L, a level once thought to be harmless. The current prevalence rates of iron deficiency and IDA in toddlers, especially among those in the lower socioeconomic groups, are unacceptably high. These young children are doubly at risk for neurodevelopmental impairment, both from the iron deficiency itself as well as from CNS damage caused by the associated increased lead absorption. The current screening and treatment recommendations for IDA in the US and in other developed countries appear to have been unsuccessful in preventing iron deficiency and IDA in a large number of toddlers. Similarly, the associated problem of impaired mental and psychomotor development has not been adequately recognized or addressed in the existing medical literature. The author recommends that, after breastfeeding or an iron-fortified formula is stopped, iron deficiency and IDA be prevented by routine daily supplemental doses of 10mg of elemental iron via iron-fortified vitamins, iron drops, or iron-fortified drinks. PMID- 16356021 TI - Controversies in the diagnosis and management of urinary tract infections in children. AB - Urinary tract infection (UTI) is one of the most common childhood bacterial infections, after upper respiratory tract and middle ear infections. The current goal of management is to prevent detrimental effects of UTI by early detection and treatment. Recommendations for the imaging of children depend upon age at presentation and sex. All children aged <5 years who have had a febrile UTI require a radiologic evaluation to identify any underlying genitourinary pathology. Older children can undergo a more tailored work-up depending on whether there is a febrile UTI or cystitis-type symptoms. Dysfunctional voiding and urge syndrome significantly increase the risk of developing UTIs in children. Vesicoureteral reflux can increase the risk of pyelonephritis and renal scarring in children with UTIs. For the most part, pyelonephritis can be diagnosed on clinical grounds in the majority of patients and a subsequent (99m)Tc dimercaptosuccinic acid scan can be reserved to identify post-nephritic renal scarring. When renal scarring is identified, the child and parents need to be educated regarding the possibility of hypertension, proteinuria, progressive nephropathy, and the risk of complications in future pregnancies. Treatment of UTI is started in the unwell child before the culture results are available and subsequently changed to culture-specific antimicrobial therapy. A short course of treatment is required for acute uncomplicated UTIs. A child with acute pyelonephritis requires 10-14 days of antibacterial treatment. The oral route in young children often causes vomiting, which implies therapeutic delay, a well known risk factor for scarring. PMID- 16356023 TI - Leukotriene receptor antagonists in children with cystic fibrosis lung disease : anti-inflammatory and clinical effects. AB - Cystic fibrosis (CF) lung disease is characterized by chronic endobronchial infection resulting in progressive pulmonary destruction; this is a major cause of mortality and morbidity. Neutrophils are the primary effector cells responsible for the progressive deterioration of lung function. Peptido leukotriene B4 antagonists, new anti-inflammatory agents that block the neutrophil-dominated inflammation, could have had the potential for long-term use. A trial on the pharmacokinetics of amelubant administered orally as a single dose of up to 75 mg in pediatric patients with CF and 300 mg in adults, and as a repeated dose of 75 mg and 150 mg, respectively, once daily for 15 days provided evidence that amelubant metabolism in adult and pediatric patients with CF is similar to that in healthy adults. In another study using the same dosage regimen, amelubant appeared to be safe and well tolerated. Safety measures included physical examination, vital signs, spirometry, oximetry, ECG, and clinical laboratory testing. However, a randomized, double-blind, placebo controlled, multinational, phase II trial (Boehringer Ingelheim 543.45) was conducted to investigate the clinical efficacy of 24 weeks of treatment with amelubant in patients with CF with mild-to-moderate lung disease. Two doses of amelubant (75 and 150 mg) were tested in adult patients (> or = 18 years) and one dose of amelubant (75mg) was tested in pediatric (6-17 years) patients. The trial was terminated early due to a statistically significant increase in the risk of pulmonary-related, serious adverse events in adults receiving amelubant. Cysteinyl leukotrienes, eosinophilic inflammation, and viral infections also contribute to progressive pulmonary destruction in CF. Cysteinyl leukotrienes are potential targets for cysteinyl leukotriene receptor antagonist use. A study on the pharmacokinetics of montelukast in children with CF provided evidence that the dose of montelukast and the administration interval does not need to be modified if the goal is to mimic the serum concentrations used to treat asthma. In a randomized, double-blind, crossover, placebo-controlled study, 16 children with mild CF (median age 9.5 years; vital capacity [VC] >70%) were treated with montelukast (5 to < or =14 years; 5 mg; >14 years; 10 mg) or placebo as a once daily tablet for 21 days. There was a significant (p < or = 0.02) reduction in serum eosinophil cationic protein levels and eosinophils (p < or = 0.027) with montelukast. However, neither lung function tests (VC, forced expiratory volume in 1 second [FEV1], maximum expiratory flow at 25% of forced VC), nor clinical symptom scores changed significantly. In another study, 26 patients aged 6-18 years with moderate CF (VC between 40% and 69% predicted) received montelukast or placebo for 8 weeks in a 20-week, randomized, double-blind, crossover, placebo controlled trial. After treatment with montelukast there was a significant improvement in FEV1, peak expiratory flow, and forced expiratory flow between 25% and 75%, and a significant decrease in cough and wheezing scale scores (p < 0.001 for all). Montelukast treatment decreased serum and sputum levels of eosinophil cationic protein and interleukin-8 (IL-8), decreased sputum levels of myeloperoxidase, and increased serum and sputum levels of IL-10 (p < 0.001 for all) compared with placebo. To date, clinical experience and research data on the anti-inflammatory effects of leukotriene receptor antagonists in CF are limited. Multicenter trials with longer observation periods and greater patient numbers are needed to prove the hypothesis that leukotriene receptor antagonists have the potential to ameliorate CF lung disease with long term use. PMID- 16356024 TI - Diagnosis and management of trichotillomania in children and adolescents. AB - Trichotillomania (TTM), a disorder characterized by the repeated non-cosmetic pulling out of hair from any part of the body, was first described in 1889. The disorder can be associated with serious social and psychologic dysfunction, as well as medical problems. A large proportion of the published scientific literature on TTM consists of case reports, and the disorder has only received significant clinical and research attention over the last 20 years. The disorder occurs across age groups and tends to follow a chronic course in the majority of cases. There is evidence for a bimodal onset, with peaks in the pre-school years and in early adolescence. TTM in child and adolescent populations has not been extensively studied, and the etiology, natural course, and best treatment approaches for the disorder are not known. Assessment for TTM in children and adolescents focuses on making the diagnosis and documenting the response to treatment. Despite the lack of validity studies in child and adolescent populations, most assessments for TTM use one or more formal TTM measures. Although classified as an impulse control disorder in the Diagnostic and Statistical Manual of Mental Disorders (4th Edition), there is some controversy about making the diagnosis in child populations because of criteria B (pattern of rising tension prior to pulling) and C (relief after pulling). There is no consensus for the treatment of TTM in children and adolescents. As in adults, a variety of interventions have been reported, including dynamic therapy, behavioral therapy, and psychopharmacology. Use of pharmacologic interventions in the pre-school age group is rare, but becomes more common as the child ages into adolescence. The most frequently used agents include clomipramine, fluoxetine, and paroxetine. The effectiveness of psychopharmacologic interventions for TTM in children and adolescents is, at best, mixed. A multiple modal approach that includes behavioral, pharmacologic, and other therapies may be the best strategy. There have been no controlled treatment trials in child and adolescent populations. Case reports favor a behavioral approach as the first-line single modality of treatment. Controlled studies of single modalities and combined treatment approaches are clearly needed. PMID- 16356025 TI - Management of Landau-Kleffner syndrome. AB - Landau-Kleffner syndrome (LKS) is an acquired epileptic aphasia disorder in which children, usually 3-8 years of age who have developed age-appropriate speech, experience language regression with verbal auditory agnosia, abnormal epileptiform activity, behavioral disturbances, and sometimes overt seizures. There are no controlled clinical trials investigating the therapeutic options for LKS. Only open-label data are available. Early diagnosis and initiation of prompt medical treatment appear to be important to achieving better long-term prognosis.Several antiepileptic drugs have been reported to be beneficial in treating this syndrome. These include valproic acid (valproate sodium), diazepam, ethosuximide, clobazam, and clonazepam. Reports on the efficacy of lamotrigine, sultiame, felbamate, nicardipine, vigabatrin, levetiracetam, vagal nerve stimulation, and a ketogenic diet are few and more experience is needed. Carbamazepine and possibly phenobarbital and phenytoin have been reported to occasionally exacerbate the syndrome. As initial therapy, valproic acid or diazepam is often empirically chosen. Subsequently, other antiepileptic drugs, corticosteroids, or intravenous immunoglobulin (IVIG) therapy are often used. Corticosteroid therapy should probably not be delayed more than 1-2 months after the initial diagnosis. Various corticosteroid regimens including oral prednisone and, recently, high doses of intravenous pulse corticosteroids, as well as corticotropin (adrenocorticotropic hormone) have been reported to be effective in LKS. Oral corticosteroids are used more often and usually need to be maintained for a long period of time to prevent relapses. The use of IVIG has been associated with an initial dramatic response in only a few patients. In our experience, a long-term worthwhile improvement has been noted in only 2 of 11 patients. These two patients had an immediate response to IVIG initially and after relapses before eventually achieving a long-term sustained remission. Surgical treatment by multiple subpial transection, which is reserved for patients who have not responded to multiple medical therapies, has been followed in selected cases by a marked improvement in language skills and behavior. However, a widely accepted consensus about suitable candidates for this surgery and about its efficacy is still lacking. Speech therapy, including sign language, and a number of classroom and behavioral interventions are helpful in managing LKS, and should be used in all patients. PMID- 16356026 TI - Pharmacotherapy of hyperlipidemia in pediatric heart transplant recipients: current practice and future directions. AB - Lipoprotein abnormalities are fairly common after pediatric heart transplantation. Graft coronary artery disease (GCAD) limits long-term survival and has been linked to elevated serum triglyceride levels and decreased high density lipoprotein levels. Histologically, GCAD represents intimal hyperplasia of the coronary vessel and is best imaged by intravascular ultrasound.A number of pharmacologic agents are available for the management of lipid disorders but experience with these drugs has mainly been in adults. HMG-CoA reductase inhibitors (statins) are currently used by many adult transplantation centers to alter lipid profiles in the hope of reducing GCAD. The use of statins among pediatric heart transplant centers is more limited. Although rhabdomyolysis is a concern with these agents, the incidence among individuals receiving immunosuppressant therapy is low. Aside from their lipid-lowering properties, statins may also protect against graft failure and rejection. PMID- 16356027 TI - Toxicokinetics of mercury elimination by succimer in twin toddlers. AB - We describe the toxicokinetics of mercury in two twin toddlers poisoned by an East Indian remedy for teething. Succimer (dimercaprosuccinic acid, DMSA) decreased the plasma elimination half-life of mercury by 3-fold in the patient with high exposure; a more modest effect was observed in the other twin. Succimer is a chelation agent used in the treatment of heavy metal intoxication. A water soluble agent, it increases the urinary excretion of lead and mercury. The drug is rapidly but variably absorbed through the gastrointestinal tract, with peak levels occurring at 1-2 hours. After its absorption, peak mercury excretion through the urine occurs within 2-4 hours. In the poisoned twin sisters, succimer administration led to a mercury plasma elimination half-life of 6 weeks. Although succimer has been used in mercury poisoning in adults and children, the toxicokinetics of mercury have not previously been characterized in either age group. We believe this is the first such description. More studies on the toxicokinetics and dynamics of mercury chelation with succimer in young children are needed to ensure the optimal use of the drug in this population. PMID- 16356028 TI - Postcollision interaction in noble gas clusters: observation of differences in surface and bulk line shapes. AB - The surface and bulk components of the x-ray photoelectron spectra of free noble gas clusters are shown to display differences in the influence of postcollision interaction between the photoelectron and the Auger electron on the spectral line shape; the bulk component is observed to be less affected than the surface and atomic parts of the spectra. A model for postcollision interaction in nonmetallic solids and clusters is also provided which takes the polarization screening into account. Core-level photoelectron spectra of Ar, Kr, and Xe have been recorded to verify the dependence of the postcollision interaction effect on the polarizability of the sample. PMID- 16356029 TI - A new method to reconstruct three-dimensional spatial distribution function from radial distribution function in solvation structure. AB - Three-dimensional spatial distribution function (SDF) of solvent is a fundamental quantity for analysis of solvation. However, its calculation has been very limited because long computational time is required. We here developed a novel and robust method to construct approximated SDFs of solvent sites from radial distribution functions. In this method, the expansion of SDFs in real solid harmonics around atoms of solute leads to a linear equation, from which SDFs are evaluated with reasonable computational time. This method is applied to the analysis of the solvation structure of liquid water, as an example. The successful results clearly show that this method is very powerful to investigate solvation structure. PMID- 16356030 TI - Static and dynamic descriptions of bond breaking/formation: a complementary view? AB - Ab initio molecular-dynamic simulations using density-functional theory and the recent atom-centered density-matrix propagation (ADMP) method were used to study the bond breaking and formation for a case-study substitution nucleophilic bimolecular reaction, namely, the Walden inversion. Using the atoms-in-molecule approach, we have performed a detailed analysis to investigate intra- and intermolecular charge transfer along the ADMP trajectory. These results were compared to those obtained considering a static approach, such as the intrinsic reaction path. In particular, the topological properties computed along the dynamic trajectory well evidence a stronger electron exchange tending to spontaneously maximize the rising covalent interaction. Furthermore, their analysis suggests that the bond formation mechanism involves a reactive intermediate with a bonding interaction stronger than in the final product. PMID- 16356031 TI - Lipid diffusion compared in outer and inner leaflets of planar supported bilayers. AB - The translational diffusion coefficient (D) of lipids located in the outer and inner leaflets of planar supported DLPC (1,2-dilauroyl-sn-glycero-3 phosphocholine) bilayers in the fluid phase was measured using fluorescence correlation spectroscopy of dye-labeled lipids at the low concentration of 0.001% and using iodide quenching of dyes in the outer leaflet to distinguish diffusion in the inner leaflet from that in the outer leaflet. To confirm the generality of these findings, the bilayers were prepared not only by vesicle fusion but also by Langmuir-Blodgett deposition. We conclude that regardless of whether the bilayers were supported on quartz or on a polymer cushion, D in the inner and outer leaflets was the same within an experimental uncertainty of +/-10% but with a small systematic tendency to be slower (by <5%) within the inner leaflet. PMID- 16356032 TI - A combined crossed-beam and theoretical investigation of radical-radical reaction dynamics of O(3P)+t-C4H9-->OH+iso-C4H8. AB - The oxidation reaction dynamics of a saturated hydrocarbon radical t-butyl leading to the isobutene +OH (X 2Pi:v"=0, 1, 2) products in the gas phase were first investigated by applying a combination of high-resolution spectroscopy in a crossed-beam configuration and ab initio calculations. By comparing the nascent OH populations with the statistical theory, the reaction mechanism at the molecular level can be described in terms of two competing dynamic pathways: the major direct abstraction process leading to the inversion of vibrational populations, and the minor short-lived addition-complex process for hot rotational distributions. PMID- 16356033 TI - Ultrasoft spin-dependent pseudopotentials. AB - The use of the spin-dependent pseudopotentials has been shown to markedly enhance the transferability of the commonly used spin-neutral pseudopotential method for the study of the structural and magnetic properties of transition-metal containing materials. Unfortunately, because the method was based on the rather expensive norm-conserving pseudopotential formalism, the method was limited to the study of fairly small systems. Here we present an extension of the spin dependent pseudopotential method for the far more computationally advantageous ultrasoft formalism and show that it is very easy to add such a feature to any preexisting computer code. We benchmark our new method by comparing to previously published results and then apply it to the study of several relevant test cases: bulk Ni, Fe, and Co, as well as a Pd atomic wire. PMID- 16356034 TI - Assessment of a new approach for the two-electron cumulant in natural-orbital functional theory. AB - The Piris natural orbital functional (PNOF) based on a new approach for the two electron cumulant has been used to predict adiabatic ionization potentials, equilibrium bond distances, and harmonic vibrational frequencies of 18 diatomic molecules. Vertical ionization potentials have been calculated for the same set of diatomic molecules and another set of 20 polyatomic molecules using energy difference methods as well as the extended Koopman theorem. The PNOF properties compare favorably with the coupled-cluster-doubles results. The calculated PNOF values are in good agreement with the corresponding experimental data, considering the basis sets used (6-31G**). PMID- 16356035 TI - Intermolecular potentials based on symmetry-adapted perturbation theory with dispersion energies from time-dependent density-functional calculations. AB - Recently, three of us have proposed a method [Phys. Rev. Lett. 91, 33201 (2003)] for an accurate calculation of the dispersion energy utilizing frequency dependent density susceptibilities of monomers obtained from time-dependent density-functional theory (DFT). In the present paper, we report numerical calculations for the helium, neon, water, and carbon dioxide dimers and show that for a wide range of intermonomer separations, including the van der Waals and short-range repulsion regions, the method provides dispersion energies with accuracies comparable to those that can be achieved using the current most sophisticated wave-function methods. If the dispersion energy is combined with (i) the electrostatic and first-order exchange interaction energies as defined in symmetry-adapted perturbation theory (SAPT) but computed using monomer Kohn-Sham (KS) determinants, and (ii) the induction energy computed using the coupled KS static response theory, (iii) the exchange-induction and exchange-dispersion energies computed using KS orbitals and orbital energies, the resulting method, denoted by SAPT(DFT), produces very accurate total interaction potentials. For the helium dimer, the only system with nearly exact benchmark values, SAPT(DFT) reproduces the interaction energy to within about 2% at the minimum and to a similar accuracy for all other distances ranging from the strongly repulsive to the asymptotic region. For the remaining systems investigated by us, the quality of the SAPT(DFT) interaction energies is so high that these energies may actually be more accurate than the best available results obtained with wave-function techniques. At the same time, SAPT(DFT) is much more computationally efficient than any method previously used for calculating the dispersion and other interaction energy components at this level of accuracy. PMID- 16356036 TI - An Aufbau Ansatz for geminal functional theory. AB - We generalize antisymmetric geminal products to more than just one generating geminal using an Aufbau Ansatz similar to the Hartree-Fock theory. Investigation of Li-, Be, B+, LiH, BeH+, and He2 shows a very high recovery of electron correlation energy using this Aufbau Ansatz. The method is inherently multideterminantal and insensitive to symmetry problems. The computational complexity is en par with configuration interaction of singles and doubles. PMID- 16356037 TI - Approximate treatment of higher excitations in coupled-cluster theory. AB - The possibilities for the approximate treatment of higher excitations in coupled cluster (CC) theory are discussed. Potential routes for the generalization of corresponding approximations to lower-level CC methods are analyzed for higher excitations. A general string-based algorithm is presented for the evaluation of the special contractions appearing in the equations specific to those approximate CC models. It is demonstrated that several iterative and noniterative approximations to higher excitations can be efficiently implemented with the aid of our algorithm and that the coding effort is mostly reduced to the generation of the corresponding formulas. The performance of the proposed and implemented methods for total energies is assessed with special regard to quadruple and pentuple excitations. The applicability of our approach is illustrated by benchmark calculations for the butadiene molecule. Our results demonstrate that the proposed algorithm enables us to consider the effect of quadruple excitations for molecular systems consisting of up to 10-12 atoms. PMID- 16356038 TI - An equation-free probabilistic steady-state approximation: dynamic application to the stochastic simulation of biochemical reaction networks. AB - Stochastic chemical kinetics more accurately describes the dynamics of "small" chemical systems, such as biological cells. Many real systems contain dynamical stiffness, which causes the exact stochastic simulation algorithm or other kinetic Monte Carlo methods to spend the majority of their time executing frequently occurring reaction events. Previous methods have successfully applied a type of probabilistic steady-state approximation by deriving an evolution equation, such as the chemical master equation, for the relaxed fast dynamics and using the solution of that equation to determine the slow dynamics. However, because the solution of the chemical master equation is limited to small, carefully selected, or linear reaction networks, an alternate equation-free method would be highly useful. We present a probabilistic steady-state approximation that separates the time scales of an arbitrary reaction network, detects the convergence of a marginal distribution to a quasi-steady-state, directly samples the underlying distribution, and uses those samples to accurately predict the state of the system, including the effects of the slow dynamics, at future times. The numerical method produces an accurate solution of both the fast and slow reaction dynamics while, for stiff systems, reducing the computational time by orders of magnitude. The developed theory makes no approximations on the shape or form of the underlying steady-state distribution and only assumes that it is ergodic. We demonstrate the accuracy and efficiency of the method using multiple interesting examples, including a highly nonlinear protein-protein interaction network. The developed theory may be applied to any type of kinetic Monte Carlo simulation to more efficiently simulate dynamically stiff systems, including existing exact, approximate, or hybrid stochastic simulation techniques. PMID- 16356039 TI - Combining fixed- and moving-grid methods to study direct dissociation processes involving nonadiabatic transitions. AB - We present a novel quantum-dynamics approach suitable for computing direct dissociation processes, including electronic transitions. This approach combines quantum trajectories in the Lagrangian reference frame with standard fixed-grid wave packets in order to overcome the limitations and difficulties of both techniques. As a model application, we consider the ultrafast photodissociation of H2 excited by a femtosecond extreme UV laser pulse. PMID- 16356040 TI - Relativistic effects on the nuclear magnetic shieldings of rare-gas atoms and halogen in hydrogen halides within relativistic polarization propagator theory. AB - In this work an analysis of the electronic origin of relativistic effects on the isotropic dia- and paramagnetic contributions to the nuclear magnetic shielding sigma(X) for noble gases and heavy atoms of hydrogen halides is presented. All results were obtained within the 4-component polarization propagator formalism at different level of approach [random-phase approximation (RPA) and pure zeroth order approximation (PZOA)], by using a local version of the DIRAC code. From the fact that calculations of diamagnetic contributions to sigma within RPA and PZOA approaches for HX(X=Br,I,At) and rare-gas atoms are quite close each to other and the finding that the diamagnetic part of the principal propagator at the PZOA level can be developed as a series [S(Delta)], it was found that there is a branch of negative-energy "virtual" excitations that contribute with more than 98% of the total diamagnetic value even for the heavier elements, namely, Xe, Rn, I, and At. It contains virtual negative-energy molecular-orbital states with energies between -2 mc2 and -4 mc2. This fact can explain the excellent performance of the linear response elimination of small component (LR-ESC) scheme for elements up to the fifth row in the Periodic Table. An analysis of the convergency of S(Delta) and its physical implications is given. It is also shown that the total contribution to relativistic effects of the innermost orbital (1s1/2) is by far the largest. For the paramagnetic contributions results at the RPA and PZOA approximations are similar only for rare-gas atoms. On the other hand, if the mass-correction contributions to sigma(p) are expressed in terms of atomic orbitals, a different pattern is found for 1s1/2 orbital contributions compared with all other s-type orbitals when the whole set of rare-gas atoms is considered. PMID- 16356041 TI - Free energy of solvation from molecular dynamics simulation applying Voronoi Delaunay triangulation to the cavity creation. AB - The free energy of solvation for a large number of representative solutes in various solvents has been calculated from the polarizable continuum model coupled to molecular dynamics computer simulation. A new algorithm based on the Voronoi Delaunay triangulation of atom-atom contact points between the solute and the solvent molecules is presented for the estimation of the solvent-accessible surface surrounding the solute. The volume of the inscribed cavity is used to rescale the cavitational contribution to the solvation free energy for each atom of the solute atom within scaled particle theory. The computation of the electrostatic free energy of solvation is performed using the Voronoi-Delaunay surface around the solute as the boundary for the polarizable continuum model. Additional short-range contributions to the solvation free energy are included directly from the solute-solvent force field for the van der Waals-type interactions. Calculated solvation free energies for neutral molecules dissolved in benzene, water, CCl4, and octanol are compared with experimental data. We found an excellent correlation between the experimental and computed free energies of solvation for all the solvents. In addition, the employed algorithm for the cavity creation by Voronoi-Delaunay triangulation is compared with the GEPOL algorithm and is shown to predict more accurate free energies of solvation, especially in solvents composed by molecules with nonspherical molecular shapes. PMID- 16356042 TI - N-K near-edge x-ray-absorption fine structures of acetonitrile in gas phase. AB - The dynamic processes of N(1s) core-hole excitation in gas-phase CH3CN molecule have been studied at both Hartree-Fock and hybrid density-functional theory levels. The vibrational structure is analyzed for fully optimized core-excited states. Frank-Condon factors are obtained using the linear coupling model for various potential surfaces. It is found that the vibrational profile of the N-K absorption can be largely described by a summation of two vibrational progressions: a structure-rich profile of nu(CN) stretching mode and a large envelope of congestioned vibrational levels related to the strong (-C-CN) terminal bending bond. Excellent agreement between theoretical and experimental spectra is obtained. PMID- 16356043 TI - Theoretical study of the adsorption of H on Si n clusters, (n=3-10). AB - A recently proposed local Fukui function is used to predict the binding site of atomic hydrogen on silicon clusters. To validate the predictions, an extensive search for the more stable SinH (n=3-10) clusters has been done using a modified genetic algorithm. In all cases, the isomer predicted by the Fukui function is found by the search, but it is not always the most stable one. It is discussed that in the cases where the geometrical structure of the bare silicon cluster suffers a considerable change due to the addition of one hydrogen atom, the situation is more complicated and the relaxation effects should be considered. PMID- 16356044 TI - Oscillator strength and linewidth measurements of dipole-allowed transitions in 14N2 between 93.5 and 99.5 nm. AB - Line oscillator strengths in 16 electric dipole-allowed bands of 14N2 in the 93.5 99.5 nm (106,950-100,500 cm(-1)) region have been measured at an instrumental resolution of 6.5 x 10(-4) nm (0.7 cm(-1)). The transitions terminate on vibrational levels of the 3psigma 1Sigma u (+), 3ppi 1Pi u, and 3ssigma 1Pi u Rydberg states and of the b' 1Sigma u (+) and b 1Pi u valence states. The J dependences of band f values derived from the experimental line f values are reported as polynomials in J'(J'+1) and are extrapolated to J'=0 in order to facilitate comparisons with results of coupled-Schrodinger-equation calculations that do not take into account rotational interactions. Most bands in this study reveal a marked J dependence of the f values and/or display anomalous P-, Q- and R-branch intensity patterns. These patterns should help inform future spectroscopic models that incorporate rotational effects, and these are critical for the construction of realistic atmospheric radiative transfer models. Linewidth measurements are reported for four bands. Information provided by the J dependences of the experimental linewidths should be of use in the development of a more complete understanding of the predissociation mechanisms in N2. PMID- 16356045 TI - Rotational effects in the band oscillator strengths and predissociation linewidths for the lowest 1Pi u-X 1Sigmag+ transitions of N2. AB - A coupled-channel Schrodinger equation (CSE) model of N2 photodissociation, which includes the effects of all interactions between the b, c, and o 1Pi u and the C and C' 3Pi u states, is employed to study the effects of rotation on the lowest upsilon 1Pi u-X 1Sigmag+(upsilon,0) band oscillator strengths and 1Pi u predissociation linewidths. Significant rotational dependences are found which are in excellent agreement with recent experimental results, where comparisons are possible. New extreme-ultraviolet (EUV) photoabsorption spectra of the key b 1Pi u<--X 1Sigmag +(3,0) transition of N2 are also presented and analyzed, revealing a b(upsilon=3) predissociation linewidth peaking near J=11. This behavior can be reproduced only if the triplet structure of the C state is included explicitly in the CSE-model calculations, with a spin-orbit constant A approximately 15 cm(-1) for the diffuse C(upsilon=9) level which accidentally predissociates b(upsilon=3). The complex rotational behavior of the b-X(3,0) and other bands may be an important component in the modeling of EUV transmission through nitrogen-rich planetary atmospheres. PMID- 16356046 TI - Ab initio and direct quasiclassical-trajectory study of the F+CH4-->HF+CH3 reaction. AB - We present an electronic structure and dynamics study of the F+CH4-->HF+CH3 reaction. CCSD(T)/aug-cc-pVDZ geometry optimizations, harmonic-frequency, and energy calculations indicate that the potential-energy surface is remarkably isotropic near the transition state. In addition, while the saddle-point F-H-C angle is 180 degrees using MP2 methods, CCSD(T) geometry optimizations predict a bent transition state, with a 153 degrees F-H-C angle. We use these high-quality ab initio data to reparametrize the parameter-model 3 (PM3) semiempirical Hamiltonian so that calculations with the improved Hamiltonian and employing restricted open-shell wave functions agree with the higher accuracy data. Using this specific-reaction-parameter PM3 semiempirical Hamiltonian (SRP-PM3), we investigate the reaction dynamics by propagating quasiclassical trajectories. The results of our calculations using the SRP-PM3 Hamiltonian are compared with experiments and with the estimates of two recently reported potential-energy surfaces. The trajectory calculations using the SRP-PM3 Hamiltonian reproduce quantitatively the measured HF vibrational distributions. The calculations also agree with the experimental HF rotational distributions and capture the essential features of the excitation function. The results of the SRP semiempirical Hamiltonian developed here clearly improve over those using the two prior potential-energy surfaces and suggest that reparametrization of semiempirical Hamiltonians is a promising strategy to develop accurate potential-energy surfaces for reaction dynamics studies of polyatomic systems. PMID- 16356047 TI - Electron-spin multiplicities and molecular structures of neutral and ionic scandium-benzene complexes. AB - Scandium-benzene complexes, Sc-(C6H6)1,2 are produced by interactions between the laser-vaporized scandium atoms and benzene vapor in pulsed molecular beams, and identified by photoionization time-of-flight mass spectrometry and photoionization efficiency spectroscopy. The electron-spin multiplicities and geometries of these complexes and their ions are determined by combining pulsed field-ionization zero electron kinetic-energy spectroscopy and density-functional theory calculations. For scandium-monobenzene, a short-range quartet ground state is determined for the neutral complex, and a low-energy triplet state is probed for the ion. For the dibenzene complex, the neutral ground state is a doublet, and two low-energy ion states are singlet and triplet. The quartet and triplet states of scandium-monobenzene and the triplet state of scandium-dibenzene possess sixfold symmetry, whereas the doublet and singlet of the dibenzene complex have twofold symmetry. Moreover, ionization energies and metal-ring stretching wavenumbers are measured for both complexes. PMID- 16356048 TI - Understanding glycine conformation through molecular orbitals. AB - The four most stable C(s) conformers of glycine have been investigated using a variety of quantum-mechanical methods based on Hartree-Fock theory, density functional theory (B3LYP and statistical average of orbital potential), and electron propagation (OVGF) treatments. Information obtained from these models were analyzed in coordinate and momentum spaces using dual space analysis to provide insight based on orbitals into the bonding mechanisms of glycine conformers, which are generated by rotation of C-O(H) (II), C-C (III), and C-N (IV) bonds from the global minimum structure (I). Wave functions generated from the B3LYP/TZVP model revealed that each rotation produced a unique set of fingerprint orbitals that correspond to a specific group of outer valence orbitals, generally of a' symmetry. Orbitals 14a', 13a', 12a', and 11a' are identified as the fingerprint orbitals for the C-O(H) (II) rotation, whereas fingerprint orbitals for the C-C (III) bond rotation are located as 16a' [highest occupied molecular orbital (HOMO)], 15a' [next highest molecular occupied molecular orbital (NHOMO)], 14a', and 12a' orbitals. Fingerprint orbitals for IV generated by the combined rotations around the C-C, C-O(H), and C-N bonds are found as 16a', 15a', 14a', 13a', and 11a', as well as in orbitals 2a" and 1a". Orbital 14a' is identified as the fingerprint orbital for all three conformational processes, as it is the only orbital in the outer valence region which is significantly affected by the conformational processes regardless rotation of which bond. Binding energies, molecular geometries, and other molecular properties such as dipole moments calculated based on the specified treatments agree well with available experimental measurements and with previous theoretical calculation. PMID- 16356049 TI - Photoinduced ion-pair formation in the (HI)m(H2O)n cluster system. AB - The temporal behavior of the photoinduced ion-pair formation process in the (HI)m(H2O)n (n=1-6 for m=1 and n=1-4 for m=2) cluster system has been studied via the coupling between the g 3Sigma- Rydberg and V 1Sigma+ valence states. Comparison of the time constants obtained to those measured in previous experiments for the analogous process in HBr-water clusters, along with a detailed analysis of the signal intensity as a function of laser-pulse power, provides new insight into and confirmation of the previously proposed ion-pair formation mechanism. PMID- 16356050 TI - Ab initio configuration-interaction investigation of optical transitions in K+He and K+H2. AB - The potassium resonance line (4s-->4p) centered around 770 nm is a major contributor to the optical extinction in the atmospheres of certain classes of brown dwarfs and extrasolar giant planets. The resonance line is significantly broadened by collisions with He and H2, and an accurate calculation of the line profile is needed for astrophysical models of these objects. As a first step, we report an accurate ab initio study of the K+He and K+H2 potential-energy curves correlating to the K 4s and 4p atomic energy levels, together with the dipole moments governing the transitions between these potential-energy curves. The molecular calculations have been carried out using a multireference configuration interaction method, with the molecular orbitals expanded in a large Gaussian basis set. The transition dipole moments show significant variation with the molecular geometry. Calculations for the K+H2 system have been carried out for a range of H2 orientations and internuclear separations, so that the effect of H2 rotation and vibration may be explicitly included in future calculations of the pressure-broadened line profiles. PMID- 16356051 TI - An anomalous x-ray diffraction study of the hydration structures of Cs+ and I- in concentrated solutions. AB - Anomalous x-ray diffraction experiments were carried out on concentrated aqueous solutions of sodium iodide (6 molal) and cesium iodide (3 molal). Data were gathered at two energies below the absorption edges of the Cs+ and I- ions in order to avoid contributions from fluorescence. The statistics and quality of the raw data were improved by the use of a focusing analyzer crystal. Differences were taken between the data sets and used to calculate the hydration structures of Cs+ and I-. The structures found are more complex than anticipated for such large ions with relatively low charge densities and show evidence of ion-pair formation in both solutions. A two-Gaussian fit to the Cs+ data gives information about the Cs+-O and Cs+-I- correlations. The central position of the Gaussian representing the Cs+-O was fixed at 3.00 A, that is, the maximum of this contribution. The other parameters were allowed to vary freely, giving a Cs+-I- distance of 3.84+/-0.05 A and coordination numbers of 7.9 and 2.7, respectively, for the Cs+-O and Cs+-I- correlations. The results on the structure of I- in the 6 molal NaI aqueous solution were also fitted to a model based on Gaussians; this gives correlations for I- -O and I- -Na+ at 3.17+/-0.06 and 3.76+/-0.06 A with respective coordination numbers of 8.8 and 1.6. The structure of I- in the 3 molal CsI solution shows overlapping contributions due to I- -H, I- -O, and I- Cs+. The best Gaussian fit gives two peaks centered at 3.00+/-0.08 and 3.82+/ 0.04 A and shows that the latter two correlations are unresolved. The hydration structures are compared with those of other alkali and halide ions. The results are also found to be in good agreement with those obtained from standard x-ray diffraction and computer simulation. PMID- 16356052 TI - Pressure tuning of the Fermi resonance in liquid methanol: implications for the analysis of high-pressure vibrational spectroscopy experiments. AB - It has been argued that pressure tuning allows for unambiguous assignment of the nonperturbed bands involved in the Fermi coupling of molecular systems in the condensed phase. Here we study the pressure evolution of the Fermi resonance occurring in liquid methanol between the symmetric methyl-stretch fundamental and the methyl-bending overtones. Our analysis is based on Raman experiments in both stretching and bending fundamental regions, which are used to evaluate the effect of pressure on accidental degeneracies occurring in the vibrational spectra of liquid methanol. We emphasize that the difference in frequency of the Fermi doublet constitutes the governing quantity to determine the condition at which the exact degeneracy of the unperturbed modes occurs. Analysis based on the intensity ratio of the Fermi doublet must be disregarded. We confirm the necessity of measuring the full vibrational spectrum under pressure in order to obtain the Fermi coupling parameters unambiguously and to give a correct assignment of the bands involved in the resonance phenomenon. We also analyze the possible occurrence of several simultaneous resonance effects using a multilevel perturbation model. This model provides an appropriate description of the frequencies observed in the experiments over the whole pressure range if we consider that the main resonance occurs between nu3 and 2nu10, in contrast to previous assignments. Our global analysis leads to some general rules concerning measurement and interpretation of high-pressure vibrational spectroscopy experiments. PMID- 16356053 TI - Kinetic theory of nucleation based on a first passage time analysis: improvement by the density-functional theory. AB - A recent kinetic theory of nucleation [see, e.g., E. Ruckenstein and B. Nowakowski, J. Colloid Interface Sci. 137, 583 (1990)] is based on molecular interactions and avoids the traditional thermodynamics. The rate of emission of molecules from a cluster is found via a first passage time analysis. This time is calculated by solving the single-molecule master equation for the probability distribution function of a surface molecule located in the potential field created by the cluster. The liquid cluster was assumed to have sharp boundaries and uniform density. In the present paper, this assumption is removed by using the density-functional theory to find the density profiles. Thus, more accurate calculations of the potential field created by the cluster, its emission rate, and nucleation rate are obtained. The modified theory is illustrated by numerical calculations for a molecular pair interaction potential combining the dispersive attraction with the hard-sphere repulsion. PMID- 16356054 TI - Spectroscopic study of water-NaCl-benzene mixtures at high temperatures and pressures. AB - Near-infrared and ultraviolet spectra of water-NaCl-benzene mixtures have been measured in the 473-573 K and 100-400 bar range and 373-498 K and 50-300 bar range, respectively. Concentrations of water in the benzene-rich phase and benzene in the water-rich phase were estimated from integrated intensities of the absorption bands. It is found that addition of NaCl in the aqueous phase suppresses transfer of water into the benzene-rich phase, and the relative decrease in water solubility in benzene exhibits good correlation with an increase in density of the aqueous NaCl solution relative to that of neat water. The salting-out constant for the water-NaCl-benzene system, which is estimated from a relative decrease in benzene solubility in the aqueous phase by addition of sodium chloride, increases significantly with increasing temperature. It is suggested that the effect of sodium chloride on the water-benzene mutual solubilities can be explained by ion-induced electrostriction of the aqueous phase. PMID- 16356055 TI - Collisional deactivation mechanism of luminescence in hydrogen-loaded Ge-doped fibers. AB - We report experimental results on the temperature and Ge-content dependencies of the decay times of the 3.1 eV emission of twofold-coordinated germaniums, excited at 3.51 eV, in both virgin and H2-loaded germanosilicate optical fibers. For T<110 K, the lifetimes are temperature invariant in the two kinds of fibers; at higher temperatures, they are progressively shortened in the H2-loaded fibers whereas they remain unaltered in the virgin fibers. On increasing the Ge content in the fiber core the lifetime decreases in the whole temperature range. We also found a direct correlation in H2-loaded samples between the dependencies on temperature of the emission lifetime and the diffusion lengths of H2 molecules in silica glass determined from earlier work. These experimental features are explained by assuming an elastic-collisional deactivation mechanism due to interaction with diffusing H2 molecules, described in terms of an Arrhenius law with activation energy approximately 64 meV that operates in competition with the normal radiative process. PMID- 16356056 TI - Chemisorption and dissociation of single oxygen molecules on Ag110. AB - The chemisorption of single oxygen molecules on Ag110 and the dissociation of the adsorbed molecules induced by tunneling electrons were studied at 13 K using a variable-low-temperature scanning tunneling microscope. Two predominant types of chemisorbed O2 molecules were identified, one with the O2 molecular axis aligned along the [001] direction of the substrate [O2(001)], and the other with the molecular axis aligned along the [110] direction [O2(110)]. Tunneling of electrons between the scanning tunneling microscope tip and O2(001) caused the molecule either to rotate or dissociate, depending on the direction of electron tunneling. In contrast, electron tunneling caused O2(110) to dissociate regardless of tunneling direction. In addition to O2(001) and O2(110), several other oxygen species and their dynamical behaviors were observed. PMID- 16356057 TI - Nematic-isotropic interfaces under shear: a molecular-dynamics simulation. AB - We present a large-scale molecular-dynamics study of nematic-paranematic interfaces under shear. We use a model of soft repulsive ellipsoidal particles with well-known equilibrium properties, and consider interfaces which are oriented normal to the direction of the shear gradient (common stress case). The director at the interface is oriented parallel to the interface (planar). A fixed average shear rate is imposed with moving periodic boundary conditions, and the heat is dissipated with a profile-unbiased thermostat. First, we study the properties of the interface at one particular shear rate in detail. The local interfacial profiles and the capillary wave fluctuations of the interfaces are calculated and compared with those of the corresponding equilibrium interface. Under shear, the interfacial width broadens and the capillary wave amplitudes at large wavelengths increase. The strain is distributed inhomogeneously in the system (shear banding), the local shear rate in the nematic region being distinctly higher than in the paranematic region. Surprisingly, we also observe (symmetry-breaking) flow in the vorticity direction, with opposite direction in the nematic and the paranematic state. Finally, we investigate the stability of the interface for other shear rates and construct a nonequilibrium phase diagram. PMID- 16356058 TI - The dynamics of solvation of an electron in the image potential state by a layer of polar adsorbates. AB - Recently, ultrafast two-photon photoemission has been used to study electron solvation at a two-dimensional metalpolar adsorbate interfaces [A. Miller et al., Science 297, 1163 (2002)]. The electron is bound to the surface by the image interaction. Earlier we have suggested a theoretical description of the states of the electron interacting with a two-dimensional layer of the polar adsorbate [K. L. Sebastian et al., J. Chem. Phys. 119, 10350 (2003)]. In this paper we have analyzed the dynamics of electron solvation, assuming a trial wave function for the electron and the solvent polarization and then using the Dirac-Frenkel variational method to determine it. The electron is initially photoexcited to a delocalized state, which has a finite but large size, and causes the polar molecules to reorient. This reorientation acts back on the electron and causes its wave function to shrink, which will cause further reorientation of the polar molecules, and the process continues until the electron gets self-trapped. For reasonable values for the parameters, we are able to obtain fair agreement with the experimental observations. PMID- 16356059 TI - The structure of ultrathin Langmuir-Blodgett films of cadmium behenate. AB - Grazing incidence x-ray-diffraction investigations of the structures of Langmuir Blodgett films of cadmium behenate with 1, 2, 3, 5, and 21 monolayers are reported. The single monolayer film, deposited on a hydrophilic substrate, showed a hexagonal structure, whereas the bilayer film, deposited on a hydrophobic substrate, had a rectangular structure with herringbone orientation of the acyl chains. With multilayer films formed on a hydrophilic substrate, it was possible to detect that the hexagonal structure of the first layer was retained when additional layers were deposited and that the additional layers had the same rectangular structure as the bilayer. PMID- 16356060 TI - Heat capacity of water in nanopores. AB - Heat capacity of controlled amounts of water in Vycor's 2 nm radius pores has been determined in real time during the course of water's isothermal nanoconfinement from bulk state at 358 K, by using temperature-modulated calorimetry. As water transfers from bulk to nanopores via the vapor phase, its heat capacity per molecule increases asymptotically toward a limiting value of 1.4 times the heat capacity of bulk water for 1.8 wt % water in Vycor and 1.04 times for 10.0 wt %. The observations indicate that vibrational and configurational contributions to the heat capacity are highest when the amount of water is insufficient to completely cover the pore wall, and they decrease as more water is present in the nanopores and water clusters form. The heat capacity of water in completely filled nanopores approaches the value for bulk water, thus indicating that the heat capacity varies with the water molecules' position in the nanopores. PMID- 16356061 TI - New isoreticular metal-organic framework materials for high hydrogen storage capacity. AB - We propose new isoreticular metal-organic framework (IRMOF) materials to increase the hydrogen storage capacity at room temperature. Based on the potential-energy surface of hydrogen molecules on IRMOF linkers and the interaction energy between hydrogen molecules, we estimate the saturation value of hydrogen sorption capacity at room temperature. We discuss design criteria and propose new IRMOF materials that have high gravimetric and volumetric hydrogen storage densities. These new IRMOF materials may have gravimetric storage density up to 6.5 wt % and volumetric storage density up to 40 kg H2/m3 at room temperature. PMID- 16356062 TI - Bond order bond polarizability model for fullerene cages and nanotubes. AB - It is still a challenge to accurately calculate the polarizabilities of large fullerene cages and nanotubes. In this paper, a simple bond order bond polarizability relationship for carbon was found, which allowed us to apply the bond polarizability model to any pentagon isolation rule (PIR) fullerene (cage or nanotube). Following this approach, the following simple equation, alpha=1.262n, was obtained relating the static dipole polarizability (alpha) of PIR fullerenes (cages or closed nanotubes) to their number (n) of carbon atoms. Furthermore, it was shown that the polarizabilities of C60 and C70, calculated on the basis of this model, are in excellent agreement with those obtained experimentally and by density-functional theory calculations. PMID- 16356063 TI - C-H activation of alkanes on Rh+ n (n=1-30) clusters: size effects on dehydrogenation. AB - The rate coefficients for the dehydrogenation of ethane, propane, and isobutane with cationic rhodium atoms Rh+ and clusters Rh+ n of up to 30 atoms were measured under single-collision conditions in a Fourier-transform ion cyclotron resonance mass spectrometer. The reaction rates are cluster size dependent and parallel for all the three alkanes. While the reactions proceed close to the theoretical collision rates for a large number of clusters, characteristic minima are observed for Rh+ (5/6/9/19/28). The degree of dehydrogenation varies with the cluster size with maxima for 10< or =n< or =15 for the three alkanes and for n=3 and 2-4 in the cases of ethane and propane, respectively. However, complete dehydrogenation is only observed for the reaction of Rh+ 11 with propane. Dehydrogenation is remarkably selective and no other neutral products than H2 are observed. The results are interpreted in terms of likely cluster geometries. PMID- 16356064 TI - Integral equation theory of randomly coupled multiblock copolymer melts: effect of block size on the phase behavior. AB - The phase behavior of randomly coupled multiblock copolymer melts is studied using the polymer reference interaction site model integral equation theory. The molecules are modeled as flexible chains with random sequences of two types of blocks, each of which consists of the same number (R) of monomer beads. In the random copolymer (R=1) limit the theory predicts macrophase separation as the temperature is decreased for all values of the monomer correlation length lambda. For R>2, however, the theory predicts a microphase separation for values of lambda less than some critical value which increases as the block size increases. PMID- 16356065 TI - Density-functional theory for fluid mixtures of charged chain particles and spherical counterions in contact with charged hard wall: Adsorption, double layer capacitance, and the point of zero charge. AB - We consider a density-functional theory to describe nonuniform fluids composed of chain molecules, containing a charged segment each, and spherical counterions. The chain molecules are modeled as freely jointed chains of hard spheres, the counterions are oppositely charged spheres of the same diameter as all segments of chain molecules. The theory is applied to study the structure of adsorbed layers, the excess adsorption isotherms, the capacitance of the double layer, and the potential of the zero charge. We show that all electric properties are strongly dependent on the length of the chain molecules. Moreover, these properties are also dependent on the position of the charged segment in the chain. PMID- 16356066 TI - Phase behavior of a blend of polymer-tethered nanoparticles with diblock copolymers. AB - Using the self-consistent field theory (SCFT), we investigate the phase behavior of a mixture of diblock copolymers and nanoparticles with monodisperse polymer chains tethered to their surfaces. We assume the size of the nanoparticles to be much smaller than that of the attached polymer chains and therefore model the particles with their grafted polymer "shell" as star polymers. The polymer chains attached to the particles are of the same species as one of the blocks of the symmetric diblock copolymer. Of primary interest is how to tune the shell of the particle by changing both the length and number of tethered polymers in order to achieve higher loading of nanoparticles within an ordered structure without macrophase separation occurring. We find that the phase behavior of the system is very sensitive to the size of the particle including its tethered shell. The region of microphase separation is increased upon decreasing the star polymer size, which may be achieved by shortening and/or removing tethered polymer chains. To explore the possible structures in these systems we employ SCFT simulations that provide insight into the arrangement of the different species in these complex composites. PMID- 16356067 TI - The microrheology of polystyrene sulfonate combs in aqueous solution. AB - Video particle tracking (VPT) and diffusing wave spectroscopy were used to characterize the microrheology of polystyrene sulfonate combs in aqueous solutions. At low frequencies VPT demonstrated predominantly viscous behavior. The manner in which the viscosity scaled as a function of monomer concentration was a sensitive function of the comb architecture. Densely branched combs or combs with long side chains demonstrated entangled polyelectrolyte scaling above the overlap concentration, whereas sparsely branched combs had unentangled polyelectrolyte scaling. A dynamic scaling model was developed for the viscosity of unentangled semidilute solutions of comb polyelectrolytes. Diffusing wave spectroscopy demonstrated Rouse modes (G' approximately G" approximately omega12) for the high-frequency dynamics of the semidilute comb solutions. The form of the high-frequency viscoelasticity was independent of the chain architecture and the modulus scaled as expected for linear flexible polyelectrolytes. PMID- 16356068 TI - Signal transduction in a coupled hormone system: selective explicit internal signal stochastic resonance and its control. AB - Cooperative interactions of signal transduction and environmental noise are investigated with a coupled hormone system, in which selective explicit internal signal stochastic resonance (EISSR) is observed. More specifically, the large peak of a period-2 oscillation (i.e., a strong signal) is greatly amplified by the environmental noise while the small peak (i.e., a weak signal) does not exhibit cooperative interactions with noise. The EISSR phenomenon could be controlled by adjusting the frequency or amplitude of an external signal and a critical amplitude for external signal is found. Significantly, the maximal signal-to-noise ratio increases almost linearly with the increment of control parameter, despite that the magnitude of the large peak is decreased. In addition, the noise does not alter the fundamental frequencies of the strong signal and the weak signal, which implicates that the system can keep its intrinsic oscillatory state and resist the effect of environmental fluctuations. PMID- 16356069 TI - Area compressibility and buckling of amphiphilic bilayers in molecular dynamics simulations. AB - The elastic modulus or area compressibility of a membrane is routinely calculated in molecular dynamics simulations as the proportionality constant relating surface tension and projected surface area. Recent studies, however, have revealed a marked system size dependence of these moduli, which we attribute to the neglect of thermal undulations in the area calculation. We discuss several methods, based on the Helfrich model and on numerical triangulation, to remedy this situation, and find a satisfying agreement between them. The Helfrich model also quantitatively describes a buckling transition observed for compressed bilayers. PMID- 16356070 TI - A reference interaction site model approach to depletion forces induced by hard rodlike particles. AB - We study the effective interactions among large hard spherical colloidal particles induced by small hard rodlike particles and compare them with those induced by small hard spherical particles to highlight the specific effects due to the anisotropic shape of the former. This is done by determining the effective pair potentials within the framework of the reference interaction site model approach. The rodlike particles are modeled as N nonoverlapping spherical units arranged in a straight line, so that their total length is N times their transversal diameter. These results are compared against those obtained in the Asakura-Oosawa limit. PMID- 16356071 TI - Calculation of the entropy of random coil polymers with the hypothetical scanning Monte Carlo method. AB - Hypothetical scanning Monte Carlo (HSMC) is a method for calculating the absolute entropy S and free energy F from a given MC trajectory developed recently and applied to liquid argon, TIP3P water, and peptides. In this paper HSMC is extended to random coil polymers by applying it to self-avoiding walks on a square lattice--a simple but difficult model due to strong excluded volume interactions. With HSMC the probability of a given chain is obtained as a product of transition probabilities calculated for each bond by MC simulations and a counting formula. This probability is exact in the sense that it is based on all the interactions of the system and the only approximation is due to finite sampling. The method provides rigorous upper and lower bounds for F, which can be obtained from a very small sample and even from a single chain conformation. HSMC is independent of existing techniques and thus constitutes an independent research tool. The HSMC results are compared to those obtained by other methods, and its application to complex lattice chain models is discussed; we emphasize its ability to treat any type of boundary conditions for which a reference state (with known free energy) might be difficult to define for a thermodynamic integration process. Finally, we stress that the capability of HSMC to extract the absolute entropy from a given sample is important for studying relaxation processes, such as protein folding. PMID- 16356072 TI - Effects of counterion valency on the damping of phonons propagating along the axial direction of liquid-crystalline DNA. AB - The phonon propagation and damping along the axial direction of films of aligned 40 wt % calf-thymus DNA rods are studied by inelastic x-ray scattering (IXS). The IXS spectra are analyzed with the generalized three effective eigenmode theory, from which we extract the dynamic structure factor S(Q,E) as a function of transferred energy E=variant Planck's over 2piomega, and the magnitude of the transferred wave vector Q. S(Q,E) of a DNA sample typically consists of three peaks, one central Rayleigh scattering peak, and two symmetric Stokes and anti Stokes Brillouin side peaks. By analyzing the Brillouin peaks, the phonon excitation energy and damping can be extracted at different Q values from about 4 to 30 nm(-1). A high-frequency sound speed is obtained from the initial slope of the linear portion of the dispersion relation below Q=4 nm(-1). The high frequency sound speed obtained in this Q range is 3100 ms, which is about twice faster than the ultrasound speed of 1800 ms, measured by Brillouin light scattering at Q approximately 0.01 nm(-1) at the similar hydration level. Our observations provide further evidence of the strong coupling between the internal dynamics of a DNA molecule and the dynamics of the solvent. The effect on damping and propagation of phonons along the axial direction of DNA rods due to divalent and trivalent counterions has been studied. It is found that the added multivalent counterions introduce stronger phonon damping. The phonons at the range between approximately 12.5 and approximately 22.5 nm(-1) are overdamped by the added counterions according to our model analyses. The intermediate scattering function is extracted and it shows a clear two-step relaxation with the fast relaxation time ranging from 0.1 to 4 ps. PMID- 16356073 TI - Comment on "a nonequilibrium molecular-dynamics method for thermal conductivities based on thermal noise" [J. Chem. Phys. 122, 081103 (2005)]. PMID- 16356074 TI - Comment on "path-integral virial estimator based on the scaling of fluctuation coordinates: application to quantum clusters with fourth-order propagators" [J. Chem. Phys. 123, 104101 (2005)]. PMID- 16356076 TI - Accurate statistical tests for smooth classification images. AB - Despite an obvious demand for a variety of statistical tests adapted to classification images, few have been proposed. We argue that two statistical tests based on random field theory (RFT) satisfy this need for smooth classification images. We illustrate these tests on classification images representative of the literature from F. Gosselin and P. G. Schyns (2001) and from A. B. Sekuler, C. M. Gaspar, J. M. Gold, and P. J. Bennett (2004). The necessary computations are performed using the Stat4Ci Matlab toolbox. PMID- 16356077 TI - Global competition dictates local suppression in pattern rivalry. AB - Certain pairs of visual patterns, when superimposed as if transparent, elicit a wavering percept in which one or the other pattern can enjoy temporary periods of exclusive visibility. This multistable perceptual phenomenon is called monocular rivalry or pattern rivalry (PR) and is most pronounced when the component patterns are faint and devoid of detail. The principal mechanisms that give rise to PR continue to be a topic of speculation. In the present study, we examine the determinants of exclusive dominance during PR using a novel stimulus in which a central portion is free of conflict. By observing the properties of suppression in this so-called rivalry-free region, we demonstrate that perception is driven largely by the global and holistic interpretation of the patterns, rather than by the need to resolve local spatial conflict. The suppression of this central region was often complete and varied as a function of the parameters of the global stimulus, including the size of the surround region, its ocular configuration, and stereoscopic depth ordering. Suppression also varied as a function of pattern continuity across the central region as well as with the temporal offset of the overlapping components. These findings demonstrate that the visibility or invisibility of a pattern is not fundamentally a product of local processing, but is instead shaped by the brain's global interpretive assumptions regarding the composition of the stimulus. PMID- 16356079 TI - Rapid detection of salient regions: evidence from apparent motion. AB - Most studies that have used Kanizsa-type illusory figures to investigate perceptual completion have treated the crisp bounding illusory contours (ICs) and the enclosed region as nondissociable stimulus attributes. However, there is evidence that enclosed "salient regions" (SRs; Stanley & Rubin, 2003) are detected even in cases when bounding ICs are not perceptually completed. Here we used apparent motion (AM) to test whether SRs are detected in the absence of crisp bounding ICs. Kanizsa-type stimuli were modified in ways that eliminated the bounding ICs, but the clear impression of an enclosed region remained. SR stimuli were embedded in an array of like inducers. On successive frames, the inducers in the array rotated in a way that resulted in translation of the enclosed region. Four speeds of translation were tested. Observers performed a two-alternative forced-choice task on the direction of translation. Perceptually completed SRs produced robust AM whether they were bound by crisp ICs or not observer performance was as good and, in certain cases, even better for SRs with no bounding ICs. We interpret these findings within a theoretical framework that makes a distinction between region-based and contour-based segmentation processes that operate in concert to achieve segmentation of the visual scene. PMID- 16356078 TI - Shared decision signal explains performance and timing of pursuit and saccadic eye movements. AB - Each voluntary eye movement provides physical evidence of a visuomotor choice about where and when to look. Primates choose visual targets with two types of voluntary eye movements, pursuit and saccades, although the exact mechanism underlying their coordination remains unknown. Are pursuit and saccades guided by the same decision signal? The present study compares pursuit and saccadic choices using techniques borrowed from psychophysics and models of response time. Human observers performed a luminance discrimination task and indicated their choices with eye movements. Because the stimuli moved horizontally and were offset vertically, subjects' tracking responses consisted of combinations of both pursuit and saccadic eye movements. For each of two signal strengths, we constructed speed-accuracy curves for pursuit and saccades. We found that speed accuracy curves for pursuit and saccades have the same shape, but are time shifted with respect to one another. We argue that this pattern occurs because pursuit and saccades share a decision signal, but utilize different response thresholds and are subject to different motor processing delays. PMID- 16356080 TI - Long-wavelength adaptation reveals slow, spectrally opponent inputs to the human luminance pathway. AB - In addition to its expected fast, additive L- and M-cone inputs (L + M), the luminance pathway has slow, spectrally opponent inputs. We have previously shown that on long-wavelength fields, the dominant slow signals change from L-M at moderate intensity levels to M-L signals at high. Here, we focus on the transition between them, which we find is marked by substantial changes in temporal phase delay, and by large and unexpected shifts in flicker spectral sensitivity. At moderate temporal frequencies, counter to the selective adaptation caused by the field, spectral sensitivity changes from being M-cone like to more L-cone-like. These changes can be accounted for by a change in the relative strengths of the slow spectrally opponent cone signals from L-M exceeding M-L below the transition to M-L exceeding L-M above it, and by the resulting changes in constructive and destructive interference between the dominant signal components. We speculate that the transition is caused by the deep-red field becoming equivalent, postreceptorally, to a green field at high bleaching levels. These results further challenge the dogma that there are separable psychophysical channels for the transmission and processing of color and luminance information. Although its output generates an achromatic percept, the luminance channel has spectrally opponent inputs. PMID- 16356081 TI - A standard model for foveal detection of spatial contrast. AB - The ModelFest data set was created to provide a public source of data to test and calibrate models of foveal spatial contrast detection. It consists of contrast thresholds for 43 foveal achromatic contrast stimuli collected from each of 16 observers. We have fit these data with a variety of simple models that include one of several contrast sensitivity functions, an oblique effect, a spatial sensitivity aperture, spatial frequency channels, and nonlinear Minkowski summation. While we are able to identify one model, with particular parameters, as providing the lowest overall residual error, we also note that the differences among several good-fitting models are small. We find a strong reciprocity between the size of the spatial aperture and the value of the summation exponent: both are effective means of limiting the extent of spatial summation. The results demonstrate the power of simple models to account for the visibility of a wide variety of spatial stimuli and suggest that special mechanisms to deal with special classes of stimuli are not needed. But the results also illustrate the limited power of even this large data set to distinguish among similar competing models. We identify one model as a possible standard, suitable for simple theoretical and applied predictions. PMID- 16356082 TI - Seeing and ballistic pointing at perisaccadic targets. AB - We studied the effects of visual references and the level of illumination on the localization of stimuli flashed briefly near the start of saccades. A translucent shutter made it possible to remove visual references, but admit light, at different times after saccadic onset. The results show that post-saccadic visual references are not necessary for compression: a consistent compression of verbally reported relative stimulus distances is found at all shutter latencies and at all post-shutter levels of illumination. They also show that positions indicated by blind pointing show no compression except when visual references remain in view for a substantial time after saccades. These results confirm that the visual system uses multiple representations of space and suggest that it weights them differently for different tasks and different viewing conditions. No single map is used exclusively for conscious perception or for motor action, and conscious perception is always subject to compression at the time of saccades. PMID- 16356084 TI - The Thr92Ala deiodinase type 2 (DIO2) variant is not associated with type 2 diabetes or indices of insulin resistance in the old order of Amish. AB - A common polymorphism of the type 2 deiodinase gene (Thr92Ala DIO2) was found to be associated with insulin resistance in a mixed Caucasian population. The aim of this study was to investigate the association of the Thr92Ala DIO2 variant to indices of insulin resistance in the Old Order Amish. A genotype-phenotype association study was performed at the research clinic in Strasburg, Pennsylvania, and the molecular genetics laboratory at the University of Maryland, Baltimore, Maryland, and the National Institutes of Health, Bethesda, Maryland. A total of 1,268 subjects participated in the Amish Family Diabetes Study. An association among the Thr92Ala DIO2 variant and type 2 diabetes, indices of insulin resistance (HOMA-IR), insulin secretion, free thyroid hormones, and thyrotropin (TSH) was found. No association was found among the Thr92Ala DIO2 variant and type 2 diabetes, impaired glucose tolerance, or body mass index (BMI) in the Amish. In nondiabetics (n = 747), the Ala92 allele tended to be associated with decreased rather then increased insulin secretion. No differences were observed in thyroid hormones or TSH. Contrary to prior findings, the Thr92Ala DIO2 variant tends to be associated with increased rather then decreased insulin sensitivity in the Amish. These findings could be secondary to a different genetic background or to environmental factors specific for this population. PMID- 16356085 TI - Gene expression profiles differ markedly in mouse strains that are (or are not) susceptible to hyperthyroidism induced using thyrotropin receptor-expressing adenovirus. AB - BALB/c mice are susceptible and C57BL/6 mice are resistant to Graves' hyperthyroidism induced by immunization with adenovirus encoding the thyrotropin receptor (TSHR) A-subunit. Both strains develop comparable levels of TSHR antibodies, but potent TSH blocking antibody activity in C57BL/6 mice likely blocks development of hyperthyroidism. We used microarrays to compare gene expression in spleens of mice immunized with A-subunit adenovirus (TSHR-Ad) or control adenovirus (Con-Ad). To preclude the effects of variable thyroxine (T(4)) levels, mice were studied when euthyroid as follows: BALB/c mice immunized three times with TSHR-Ad or Con-Ad and C57BL/6 mice immunized three times with TSHR-Ad or Con-Ad. Among the 14,000 expressed probe sets, there were no statistically significant differences in gene expression in BALB/c mice immunized with TSHR-Ad versus Con-Ad. In contrast, expression of 57 transcripts (representing 40 genes) changed in response to TSHR-Ad in C57BL/6 mice. Diverse genes were identified, including proteins involved in immune responses, inflammation, and cell cycling as well as heat-shock proteins and proteases. Down-regulation of chitinase 3- and 4 gene expression likely reflects cytokines produced by T-helper 2 (Th2) type cells. Indeed, the immunoglobulin (IgG) subclass for TSHR antibodies reflects a deviation away from Th2 cytokines and toward Th1 in C57BL/6 mice. In conclusion, TSHR-Ad immunization altered gene expression profiles in C57BL/6, but not in BALB/c, mice. This response primarily involved reduced gene expression. In C57BL/6 mice, decreased expression of genes such as cathelicidin, calgranulins, and lipocalin following TSHR A-subunit adenovirus immunization suggests the importance of innate immunity in this response. PMID- 16356086 TI - The possible contribution of anti-Gal to Graves' disease. AB - Anti-Gal is a natural antibody specific for the alpha-galactosyl epitope. Previous studies suggested that Graves' disease (GD) patients had elevated anti Gal titers compared to normal controls, but titers returned to normal after treatment. We developed an anti-Gal enzyme-linked immunosorbent assay (ELISA) using the property of anti-Gal to bind tightly to mouse laminin. We found no significant correlations between anti-Gal and thyroidstimulating immunoglobulin (TSI) or free thyroxine (T(4)) in untreated hyperthyroid GD patients (n = 15) without clinical ophthalmopathy or euthyroid, previously treated GD patients with ophthalmopathy. There was a significant regression between TSI and free T(4) in the hyperthyroid patients (p < 0.01). Addition of total anti- Gal antibody to the regression showed a trend toward improved correlation (p = 0.15 for improved correlation relative to TSI and free T(4) alone), suggesting it may stimulate GD thyroid tissue. However, in contrast to previous studies we found hyperthyroid patients (n = 20) had lower levels of anti-Gal immunoglobulin G (IgG) (18.4 +/- 4.0 vs. 41.8 +/- 8.9) than normals (n = 36 p < 0.05). Interestingly, hyperthyroid patients without clinical ophthalmopathy tended to have lower IgG anti-Gal levels than euthyroid patients with ophthalmopathy (p = 0.1). Hyperthyroidism significantly lowers anti-Gal, but the possible increase of anti-Gal in patients with ophthalmopathy suggests anti-Gal may play a role in ophthalmopathy, or may reflect the euthryoid status of these patients. This trend needs further study. PMID- 16356087 TI - Pre-incision infiltration of local anesthetic reduces postoperative pain with no effects on bruising and wound cosmesis after thyroid surgery. AB - Optimizing postoperative pain control is an important aspect in perioperative patient care. The aim of this study was to investigate the efficacy of preincision local anesthetic infiltration in postoperative pain management for thyroid surgery and its relationship to bruising and wound cosmesis. In a randomized single-blinded study, 39 consecutive patients listed for thyroid surgery were assigned into two groups. Group I (n = 19) received subcuticular preincision infiltration with 10 ml of bupivacaine (0.5%) and Group II (n = 20) received no infiltration. Postoperatively, the pain experienced was evaluated by two methods: verbal response scores and linear analogue scores (0-100 mm) at different time intervals following surgery. Bruising and cosmetic effects resulting from surgery were assessed using a linear analogue score at discharge. The two groups were well matched for confounding variables. Pain scores were significantly different at 6 hours post operatively (p = 0.0341) with mean scores Group I = 33 and Group II = 50, but this difference disappeared at 24 hours. No patients (0%) received IV morphine in Group I compared to 5 patients (25%) in Group II. There was no significant difference in the mean bruising scores (p = 0.8864) and mean cosmetic scores (p = 0.3339) at discharge. Preincision infiltration with bupivacaine provides easy and better analgesic control postoperatively in patients following thyroid surgery with no effects on bruising or wound cosmesis. PMID- 16356088 TI - Measurement of thyroid blood flow area is useful for diagnosing the cause of thyrotoxicosis. AB - We have utilized color Doppler ultrasonography (CDU) to evaluate the thyroid blood flow area (TBFA) quantitatively, and we propose criteria to differentiate Graves' disease (GD) and destruction-induced thyrotoxicosis (DT) in patients with thyrotoxicosis. We studied 32 patients with diffuse toxic goiter, 21 with GD in the euthyroid state, 12 with chronic thyroiditis in the euthyroid state, and 31 normal individuals. TBFA was calculated as (thyroid blood flow area/thyroid area) x 100%. CDU showed high sensitivity (84%) and specificity (90%) in distinguishing GD from DT when TBFA was between 7.7% and 8.8%. Using CDU to diagnose GD in cases with TBFA >or=8% or positive serum anti-thyrotropin receptor antibody (TRAb), the sensitivity was 95% and the specificity was 90%, which are similar results to those obtained when GD was diagnosed by radioactive iodine uptake (sensitivity 100%, specificity 90%). Therefore, CDU is a more useful and economical method of distinguishing GD patients with TBFA of 8% or above from DT than measurement of TRAb or radioactive iodine uptake. PMID- 16356089 TI - Hypothyroidism might be related to breast cancer in post-menopausal women. AB - An association between breast cancer and thyroid (autoimmune) diseases or the presence of thyroid peroxidase antibodies (TPOAb; a marker of thyroid autoimmune disease) has been suggested. However, little is known about whether women with thyroid (autoimmune) diseases are at increased risk for developing breast cancer. This cross-sectional and prospective cohort study investigated whether the presence of TPOAb or thyroid dysfunction is related to the presence or development of breast cancer. An unselected cohort of 2,775 women around menopause was screened for the thyroid parameters thyrotropin (TSH), free thyroxine (FT(4)), and TPOAb during 1994. Detailed information on previous or actual thyroid disorders and breast cancer, and on putative factors related to breast cancer and thyroid disorders, was obtained. Clinical thyroid dysfunction was defined by both abnormal FT4 and TSH, and subclinical thyroid dysfunction by abnormal TSH (with normal FT4). A TPOAb concentration >or= 100 U/ml was defined as positive (TPOAb(+)). The study group was linked with the Eindhoven Cancer Registry to detect all women with (in situ) breast cancer (ICD-O code 174) diagnosed between 1958 and 1994. Subsequently, in the prospective study, all women who did not have breast cancer in 1994 (n = 2,738) were followed up to July, 2003, and all new cases of (in situ) breast cancer and all cancer-related deaths were registered. Of the 2,775 women, 278 (10.0%) were TPOAb(+). At the 1994 screening, 37 women (1.3%) had breast cancer. TPOAbs were (independently) related to a current diagnosis of breast cancer (OR = 3.3; 95% CI 1.3-8.5). Of the remaining women, 61 (2.2%) developed breast cancer. New breast cancer was related to: (1) an earlier diagnosis of hypothyroidism (OR = 3.8; 95% CI 1.3 10.9); (2) the use of thyroid medication (OR = 3.2; 95% CI 1.0-10.7); and (3) low FT4 (lowest tenth percentile: OR = 2.3; 95% CI 1.2-4.6). In the first 3 years follow up, the relationship between FT4 and log-TSH was disturbed in women with a new breast cancer diagnosis. The presence of TPOAb was not related to breast cancer during follow-up. A direct relationship between thyroid autoimmunity and breast cancer is unlikely. Hypothyroidism and low-normal FT4 are related with an increased risk of breast cancer in post-menopausal women. Studies are needed to clarify the origins of this possible association. PMID- 16356090 TI - Fractionated dosage of radioiodine for the ablation of differentiated thyroid carcinoma. AB - In some countries with a limited number of specialized hospital beds for radionuclide therapy, ablation therapy (RIT) of differentiated thyroid carcinoma (DTC) is performed using a fractionated dosage of radioiodine. The aim of this study was to evaluate the early clinical outcome of ablation with fractionated doses of RIT in comparison to the ablation with a single dose. A subset of 386 subjects with DTC referred for the initial RIT was selected retrospectively for the study. Of these, 113 patients (29.3%) were treated with one (131)I dose of 2.2 GBq (group 1, RIT between 2001 and 2003) and 273 patients (70.7%) with fractionated doses (1.1 GBq + 1.1 GBq administered in 24 hour intervals) (group 2, RIT between 1999 and 2001). The early outcome of the initial RIT was evaluated 6-8 months later by radioiodine uptake test (RIU), thyroglobulin concentration, whole-body diagnostic scan, and neck ultrasound. On the basis of these results, the patients were classified as: CR, complete remission; NCR, no complete remission. Frequency of CR and NCR outcomes and the parameters measured during the follow-up evaluation in both groups were compared. CR outcome was found in 69 patients (61.1%) of group 1 and in 172 patients (63.0%) of group 2 (p = n.s.). No difference in measured parameters was found in both groups at the follow-up evaluation. In uncomplicated cases of DTC, RIT using a regimen of a fractionated dosage, is equally effective as the therapy with a single dose. No influence of stunning was observed in patients treated with a fractionated dosage, but the time interval between the doses was 24 hours. PMID- 16356091 TI - Early diagnosis and curative therapy of medullary thyroid carcinoma by routine measurement of serum calcitonin in patients with thyroid disorders. AB - To identify patients with medullary thyroid carcinoma (MTC) at a potentially curable stage of the disease, serum concentrations of calcitonin (hCT) were determined in 14,000 patients (including 10,158 patients with thyroid nodules) referred to a thyroid outpatient clinic. Excluding patients in whom elevated basal hCT concentrations had already been known at the time of their referral, 507 patients with thyroid nodules presented basal concentrations of hCT of more than 10 pg/ml. Following stimulation by IV pentagastrin (0.5 microg/kg BW), hCT concentrations of more than 100 pg/ml were seen in 103 patients. This group included 32 new cases of MTC (29 patients with sporadic MTC and 3 new index cases of the familial form) and 43 patients with C cell hyperplasia (CCH). Among the 3,843 patients without thyroid nodules, 2 were found to harbor sporadic MTC while 4 had CCH. As compared to 1.1 cases of MTC per 1,000 patients with nodular thyroid diseases diagnosed in our institution before hCT screening was begun, 3.2 cases of MTC per 1,000 patients were identified when hCT was determined in all patients with thyroid nodules. The determination of hCT in all patients with thyroid nodular disease facilitates the timely diagnosis of MTC, thus providing the chance of curative surgery. PMID- 16356092 TI - Putative association between anti-Borrelia IgG and autoimmune thyroid disease? AB - An association between borreliosis and autoimmune thyroid disease (AITD) has previously been discussed. We undertook the present analyses to explore associations between anti-Borrelia immunoglobulin G (IgG) and AITD using data from the population-based Study of Health in Pomerania (SHIP), which was conducted in a region with endemic Borreliosis. Data of 4,256 persons were available for the present analyses. IgG antibodies to Borrelia were determined by enzyme-linked immunosorbent assay (ELISA). AITD was defined as the combined presence of a hypoechogenic thyroid pattern in thyroid ultrasound and positive anti-thyroperoxidase levels. An association between anti-Borrelia IgG and AITD was neither present in bivariate nor in multivariate statistical analyses. This result was not affected by using different definitions of the exposure and the end points. There is no association between the exposure to Borrelia as defined by anti-Borrelia IgG and the risk of AITD. It seems unlikely that Borrelia species represent a relevant causal factor for AITD. PMID- 16356093 TI - Graves' disease: a long-term quality-of-life follow up of patients randomized to treatment with antithyroid drugs, radioiodine, or surgery. AB - The effects of treatment modality for Graves' disease (GD) were studied with respect to long-term quality of life and present health status. A total of 179 patients with GD were randomized during the period 1983-1990 for treatment with antithyroid drugs, radioiodine, or surgery. A 36-item Short Form Health Status Survey questionnaire and specific questions for GD were sent to patients 14-21 years after randomization. Present medical records, and clinical and laboratory status were recorded. No major significant differences in quality of life among the three treatments were observed. Compared to a large Swedish reference group, all treatment groups had significantly lower scores for vitality (p < 0.05). The Mental Component Summary was lower for both the young medical, young surgical, and the older medical group (p < 0.05). Radioiodine-treated patients had a lower General Health score. Young medical patients (<35 years) had lower Mental Health scores (p < 0.05). There was also a strong trend, which barely met statistically significance, for older surgical and radioiodine groups for lower Mental Component Summary. GD patients have, compared with a large Swedish reference population, diminished vital and mental quality of life aspects even many years after treatment. The quality-of-life scores were not different among the three treatment modalities. PMID- 16356094 TI - Increased risk of Graves' disease after pregnancy. AB - The improvement in autoimmune thyroid disease during pregnancy and the subsequent exacerbation postpartum is secondary to immune system changes necessary to a normal pregnancy. Prior studies have shown that a clinically significant number of women develop Graves' disease (GD) in the postpartum period. The aim of this study was to examine the risk of post pregnancy GD and define patient characteristics that may impact the diagnosis and treatment strategies for this group. We performed a retrospective review of 152 consecutive women, aged 18-39 years when diagnosed with GD, to examine the relation between disease diagnosis and prior pregnancy. Differences in patient characteristics and treatment outcomes of women were analyzed. New York City population data were used to estimate a relative risk for the development of postpartum GD. We found that in parous women, 45% were diagnosed with GD in the postpartum period and 55% had an onset in subsequent years. No significant differences were noted in patient characteristics or treatment outcomes. We found that the risk of women developing post pregnancy GD was greatest in the older patients (35-39 years), with 56% developing GD compared to 42% of nulliparous women. These data, therefore, suggest an increased risk for older women. We were able to calculate the increase in estimated relative risk for postpartum disease by using control population data. The risk maximized at 5.6 for the age group 35-39 years when compared to the control population. These data support earlier studies that showed that a clinically significant number of women develop GD after childbirth compared to nulliparous women and extends this risk for many years. The mechanism of this long-standing increased susceptibility requires further delineation. PMID- 16356095 TI - Iodine excretion in regions of Ukraine affected by the Chornobyl Accident: experience of the Ukrainian-American cohort study of thyroid cancer and other thyroid diseases. AB - Urinary iodine concentrations were measured in 11,926 subjects who are participants in the Ukrainian-American Cohort Study of Thyroid Cancer and Other Thyroid Diseases Following the Chornobyl Accident. Measurements were made in two time periods corresponding to the first and second thyroid screening cycles (1998 2000, 2001-2003). These time periods fall before and after initiation of a government program to increase iodine sufficiency. Median urinary iodine concentrations did increase in the later time period compared to the earlier [47.5 microg/L, 95% confidence interval (CI) 46.5-48.9 microg/L versus 41.7 microg/L, 95% CI 40.4-42.5 microg/L], but levels remained in the mild-to-moderate iodine deficiency range as defined by the World Health Organization (WHO), indicating the need for further efforts at iodination. In both time periods, urinary iodine levels were found to vary by place of residence and were lower in rural compared to urban areas. Iodine status needs to be considered when evaluating risk of thyroid cancer and other thyroid diseases. PMID- 16356096 TI - Definitive treatment for persistent hypoparathyroidism in a kidney transplant patient: parathyroid allotransplantation. AB - Post-surgical hypocalcemia is usually a transitory complication in thyroid and parathyroid surgery that can be resolved quickly, although it becomes a delicate matter when the problem persists. Parathyroid transplantation is the choice of treatment; however, the associated immunosuppression can cause side effects. The following case study shows the transplantation of parathyroid tissue from a patient with secondary hyperparathyroidism to another kidney transplant patient with severe hypocalcemia that was medically intractable. The graft is functioning after 2 years. PMID- 16356097 TI - Primary hyperparathyroidism as the leading symptom in a patient with a Y791F RET mutation. AB - Primary hyperparathyroidism (PHP; serum calcium 2.75 mmol/L, PTH 226 pg/ml) had been the first clinical manifestation of MEN-2A in a female patient (aged 55 years) with a mutation (Y791F, TAT-->TTT) in exon 13 of the RET proto-oncogene. The patient has a pentagastrin-induced rise in serum calcitonin (up to 57 pg/ml) considered normal for noncarriers but abnormal in family members of MEN-2 patients. This is the first case of MEN-2 due to this specific mutation with primary hyperparathyroidism as the first manifestation of the disease. In addition, the patient harbored, within the Menin gene, a polymorphism (D418D) reportedly associated with sporadic primary hyperparathyroidism. This case report indicates that molecular biological tests in MEN- 2 may only suggest a certain phenotype but cannot predict it with certainty. It may also suggest that genetic screening for MEN-2 may be advisable in patients with primary hyperparathyroidism and a borderline-high pentagastrin stimulation test, even in the absence of a positive family history. PMID- 16356098 TI - Hurthle cell carcinoma metastatic to uterus. PMID- 16356099 TI - Uncommon case of orbital metastasis secondary to papillary thyroid carcinoma. PMID- 16356101 TI - Alert: 6-mercaptopurine may be erroneously dispensed instead of propylthiouracil. PMID- 16356103 TI - Emerging roles of reactive oxygen and nitrogen species in stem/progenitor cells. PMID- 16356104 TI - Nitric oxide synthases modulate progenitor and resident endothelial cell behavior in galactosemia. AB - We used knockout animals of either inducible nitric oxide synthase (iNOS(/)) or endothelial NOS (eNOS(/)) to characterize the role of NOS in galactosemia, a model of diabetic retinopathy. NADH oxidase and nitrotyrosine were used as biomarkers of oxidative stress and vascular dysfunction. These animals were engrafted with hematopoietic stem cells (HSC) expressing green fluorescence protein (gfp(+)) to characterize the contribution of HSC and endothelial progenitor cells to neovascularization. Increased NADH oxidase activity and superoxide generation occurred in all galactose-fed mice. eNOS(/) mice demonstrated increased iNOS immunoreactivity in their retinal vasculature. Nitrotyrosine levels were low at baseline in the wild-type (WT) mice, eNOS(/) and iNOS(/) mice, and the galactose-fed iNOS mice and increased following galactose feeding in eNOS(/) and WT. Galactose-fed WT.gfp and iNOS(/).gfp chimeric animals had areas of perfused new vessels composed of gfp(+) cells. In contrast, galactose-fed eNOS(/).gfp mice produced copious, unbranched, nonperfused tubes. Thus, nitric oxide modulates HSC behavior and vascular phenotype in the retina. Although there is increased NADH oxidase and superoxide in galactosemic mice of all isoforms, iNOS is the source of nitric oxide responsible for peroxynitrite and nitrotyrosine formation that leads to the pathology observed in galactosemic mice. PMID- 16356105 TI - Reactive oxygen species as signaling molecules in cardiovascular differentiation of embryonic stem cells and tumor-induced angiogenesis. AB - Besides the well known pathophysiological impact of oxidative stress in cardiovascular disease, reactive oxygen species (ROS) generated at low concentrations exert a role as signaling molecules that are involved in signal transduction cascades of numerous growth factor-, cytokine-, and hormone-mediated pathways, and regulate biological effects such as apoptosis, cell proliferation, and differentiation. Embryonic stem cells have the capacity to differentiate into the cardiovascular cell lineage. Furthermore, upon confrontation culture with tumor tissue, they form blood vessel-like structures that induce tumor-induced angiogenesis within tumor tissues. The role of ROS in cardiovascular differentiation of embryonic stem cells appears to be antagonistic. Whereas continuous exposure to ROS results in inhibition of cardiomyogenesis and vasculogenesis, pulse chase exposure to low-level ROS enhances differentiation toward the cardiomyogenic as well as vascular cell lineage. This review summarizes the current knowledge of ROS-induced cardiovascular differentiation of embryonic stem cells as well as the role of ROS in tumor-induced angiogenesis. PMID- 16356106 TI - Role of Rac-GTPase and reactive oxygen species in cardiac differentiation of stem cells. AB - In the life of a cell, there is a constant balance between generation of reactive oxygen species (ROS) and activity of antioxidant defense mechanisms. Besides the damaging effects of ROS on many biomolecules, ROS also play a significant role in signal transduction pathways of growth factors suggesting a role of oxidative species in cell differentiation. ROS have recently been involved in the process of cardiac differentiation of stem cells. Several molecular mechanisms, including ones mediated by the GTPase Rac that underlie the regulatory role of ROS in the process of stem cell differentiation toward a cardiac lineage, are reviewed. PMID- 16356107 TI - Rho GTPases in hematopoietic cells. AB - The ubiquitous Rho GTPases are instrumental in the organization of the actin cytoskeleton, but also for the control of gene expression. Here we review the role of the major members of this family, i.e., RhoA, Rac1, Rac2, and Cdc42, and their intracellular signaling in hematopoietic cells. Although these proteins have been classically implicated in chemotaxis, there are now clear indications on how differential signaling toward other, more specific functions, such as phagocytosis or the production of reactive oxygen species, is regulated by relatively small differences in primary sequence. The identification of mutations in these GTPases or their regulators has provided novel insights in their function as well as their relevance for the development of hematological diseases. PMID- 16356108 TI - Redox regulation of precursor cell function: insights and paradoxes. AB - Studies on oligodendrocytes, the myelin-forming cells of the central nervous system, and on the progenitor cells from which they are derived, have provided several novel insights into the role of intracellular redox state in cell function. This review discusses our findings indicating that intracellular redox state is utilized by the organism as a means of regulating the balance between progenitor cell division and differentiation. This regulation is achieved in part through cell-intrinsic differences that modify the response of cells to extracellular signaling molecules, such that cells that are slightly more reduced are more responsive to inducers of cell survival and division and less responsive to inducers of differentiation or cell death. Cells that are slightly more oxidized, in contrast, show a greater response to inducers of differentiation or cell death, but less response to inducers of proliferation or survival. Regulation is also achieved by the ability of exogenous signaling molecules to modify intracellular redox state in a highly predictable manner, such that signaling molecules that promote self-renewal make progenitor cells more reduced and those that promote differentiation make cells more oxidized. In both cases, the redox changes induced by exposure to exogenous signaling molecules are a necessary component of their mode of action. Paradoxically, the results obtained through studies on the oligodendrocyte lineage are precisely the opposite of what might be predicted from a large number of studies demonstrating the ability of reactive oxidative species to enhance the effects of signaling through receptor tyrosine kinase receptors and to promote cell proliferation. Taken in sum, available data demonstrate clearly the existence of two distinct programs of cellular responses to changes in oxidative status. In one of these, becoming even slightly more oxidized is sufficient to inhibit proliferation and induce differentiation. In the second program, similar changes enhance proliferation. It is not yet clear how cells can interpret putatively identical signals in such opposite manners, but it does already seem clear that resolving this paradox will provide insights of considerable relevance to the understanding of normal development, tissue repair, and tumorigenesis. PMID- 16356109 TI - Endothelial progenitor cell dysfunction in type 1 diabetes: another consequence of oxidative stress? AB - Endothelial progenitor cells (EPC) have been shown to contribute to neovascularization and vascular maintenance and repair in adults. Recently, the concept has evolved that EPC dysfunction, in patients at risk for cardiovascular disease, may contribute to the development of atherosclerosis and ischemic vascular disease. Particularly, patients with diabetes mellitus are likely to be affected by EPC dysfunction as several studies have shown a reduced number and function of EPC in patients, as well as in preclinical models for type 1 diabetes. Here, we review our current understanding of EPC (dys)function in diabetes and discuss some potential mechanisms underlying their altered properties. Moreover, we provide circumstantial evidence indicating that increased oxidative stress could play a role in the development of EPC dysfunction in type 1 diabetes. Finally, we discuss the potential implication of our findings for EPC-based therapies and the potential impact of pharmacological interventions on the vascular regenerative capacity of EPC. PMID- 16356111 TI - Redox-based mechanisms in diabetes. PMID- 16356110 TI - Hyperglycemia-induced reactive oxygen species and impaired endothelial progenitor cell function. AB - Vascular complications in diabetes are a significant source of human morbidity and mortality, affecting multiple organ systems and persisting despite tight glucose control. Many of these complications can be linked to impairments in vasculogenesis, the process by which circulating and bone marrow-derived endothelial progenitor cells (EPCs) contribute to new vessel formation. Recent evidence suggests that hyperglycemia alone, through the mitochondrial overproduction of reactive oxygen species (ROS), can induce changes in gene expression and cellular behavior in diabetes. In this review, we examine how hyperglycemia-induced overproduction of ROS could explain EPC impairments observed in diabetes. Experimentally, impairments in EPC function prevent new blood vessel growth and are potentially reversible by manipulations to decrease ROS. Novel strategies aimed at reducing hyperglycemia-induced ROS may be a useful adjuvant to antihyperglycemic therapies in the restoration of vasculogenesis and the prevention of diabetic complications. PMID- 16356112 TI - Bovine retinal pericytes are resistant to glucose-induced oxidative stress in vitro. AB - Diabetic retinopathy is a sight-threatening complication of diabetes, and loss of pericytes represents early signs of its development. We tested the hypothesis that high glucose levels may induce signs of oxidative stress in cultured bovine retinal pericytes. Pericytes were exposed to either normal (5.5 mM) or high (22 mM) glucose levels for 1, 3, and 5 days. Signs of oxidative stress were measured by expression of copper/zinc superoxide dismutase, manganese superoxide dismutase, catalase, and glutathione peroxidase using real-time RTPCR. To elucidate the role of oxidative stress, we also measured glutathione (GSH) concentration in the cells and investigated the impact of thiol-reactive metal ions and hydrogen peroxide (H(2)O(2)) on intracellular GSH. Despite the stimulation with high glucose, thiol-reactive metal ions, or H(2)O(2), there was no clear increased expression of antioxidant enzymes or influence of GSH levels. Lipid peroxidation (malondialdehyde level) was increased in bovine aortic smooth muscle cells, but not in bovine retinal pericytes. The data indicate that pericytes do not develop oxidative stress in response to hyperglycemia. However, it is not definitively excluded that oxidative stress may occur after longer time periods of glucose stimulation. PMID- 16356113 TI - Cell culture modeling to test therapies against hyperglycemia-mediated oxidative stress and injury. AB - The concept that oxidative stress is a key mediator of nerve injury in diabetes has led us to design therapies that target oxidative stress mechanisms. Using an in vitro model of glucose-treated dorsal root ganglion (DRG) neurons in culture, we can examine both free radical generation, using fluorimetric probes for reactive oxygen species, and cell death via the TUNEL assay. The cell culture system is scaled down to a 96-well plate format, and so is well suited to high throughput screening. In the present study, we test the ability of three drugs, nicotinamide, allopurinol, and alpha-lipoic acid, alone and in combination to prevent DRG neuron oxidative stress and cell death. This combination of drugs is currently in clinical trial in type 1 diabetic patients. We demonstrate independent effects on oxidative stress and neuronal survival for the three drugs, and neuronal protection using the three drugs in combination. The data strengthen the rationale for the current clinical trial. In addition, we describe an effective tool for rapid preclinical testing of novel therapies against diabetic neuropathy. PMID- 16356114 TI - Glucose-induced enhancement of hemin-catalyzed LDL oxidation in vitro and in vivo. AB - Growing evidence indicates that oxidative modification of low-density lipoprotein (LDL) is increased in diabetes mellitus; however, the mechanism(s) of this phenomenon is still unclear. gamma-Glutamyl semialdehyde (gammaGSA) is a product of hemin (Fe(3+)-protoporphyrin IX)-catalyzed oxidation of apolipoprotein B-100 (apoB- 100) proline and arginine residues. On reduction, gammaGSA forms 5-hydroxy 2-aminovaleric acid (HAVA). This report describes the application of sensitive HAVA assay, to characterize gammaGSA formation in LDL under normo- and hyperglycemic conditions, both in vitro and in vivo. In vitro studies revealed that apoB-100 proline and arginine residues are not oxidized to HAVA by HOCl or the myeloperoxidase/hydrogen peroxide (H(2)O(2)) oxidation system. Cu(2+), Cu(2+)/H(2)O(2), and Fe(2+) induced only minor HAVA formation. In contrast, the hemin oxidation system appeared reactive toward LDL apoB-100 proline and arginine residues. The resulting significant HAVA formation was specifically inhibited by a redox-inert ferric iron chelator. Glucose further enhanced hemin-induced increase in relative electrophoretic mobility of LDL and apoB-100 HAVAformation. In vivo we observed elevated concentrations of HAVA in LDL apoB-100 in patients with impaired glucose tolerance and with manifest diabetes mellitus. In conclusion, glucose promotes iron-mediated oxidation of apoB- 100 proline and arginine residues via a superoxide-dependent mechanism, thus rendering the LDL particles more atherogenic. The findings (a) identify a potential mechanism of enhanced atherogenesis in subjects with diabetes mellitus and (b) support the value of HAVA as a specific marker of LDL apoB-100 oxidation. Antioxid. Redox Signal. 7, 1507-1512. PMID- 16356115 TI - Ischemia-reperfusion injury causes oxidative stress and apoptosis of Schwann cell in acute and chronic experimental diabetic neuropathy. AB - Mild ischemia-reperfusion (IR) injury to diabetic peripheral nerve is known to cause severe ischemic fiber degeneration. Little information is available on its effects on Schwann cell (SC). In this study, we evaluated oxidative stress and apoptosis of SC following mild IR, using immunohistochemistry in streptozotocin (STZ)- induced diabetic rats. Twenty-six rats were divided into four groups according to the duration of diabetes: 1- month STZ-induced diabetic group (n=7) and age-matched control group (n=7); 4-month STZ-induced diabetic group (n=6) and age-matched control group (n=6). Using our established IR model of 3 h of ischemia followed by 7 days of reperfusion, sciatic and tibial nerves were harvested and labeled with 8-hydroxydeoxyguanosine (8-OHdG; oxidative stress marker), caspase-3 (apoptotic executor), and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) activity (apoptotic indicator). Marked positive staining with 8-OHdG, caspase-3, and TUNEL were found in diabetic ischemic nerves (right side) following IR in both 1-month and 4-month groups. Only mild positive staining or no staining was seen in the nonischemic side (left side) of diabetic and age-matched control groups. Co-labeling with S 100 confirmed that the cells labeled with 8-OHdG, caspase3, and TUNEL were SC. SC was susceptible to oxidative injury and apoptosis in experimental diabetic neuropathy when subjected to mild IR injury. PMID- 16356116 TI - Peripheral nerve dysfunction in experimental diabetes is mediated by cyclooxygenase-2 and oxidative stress. AB - Glucose-mediated oxidative stress and alterations in cyclooxygenase (COX) pathway activity with secondary deficits of endoneurial perfusion have been implicated in the pathogenesis of experimental diabetic neuropathy (EDN). We have previously reported that activation of the COX-2 pathway is an important mediator of neurochemical and neurovascular defects in EDN in a rat model. Considering that chemical COX inhibition may exert other pharmacological effects in addition to inhibition of COX activity, the aim of this study was to explore the role of COX 2 in experimental diabetic neuropathy, using a COX-2 knockout mouse model. Here we provide evidence that COX-2 inactivation had a protective effect against diabetes-induced motor and sensory nerve conduction slowing and impaired nerve antioxidative defense that were clearly manifest in the wild-type (COX-2(+/+)) diabetic mice. These preliminary data support the role of the activation of the COX-2 pathway in mediating sensory and motor nerve conduction velocity deficits in EDN. These findings also suggest that the COX-2 pathway seems to be an important modulator of oxidative stress in EDN. PMID- 16356117 TI - Regulation of the human taurine transporter by oxidative stress in retinal pigment epithelial cells stably transformed to overexpress aldose reductase. AB - In diabetes, overexpression of aldose reductase (AR) and consequent glucose induced impairment of antioxidant defense systems may predispose to oxidative stress and the development of diabetic complications, but the mechanisms are poorly understood. Taurine (2-aminoethanesulfonic acid) functions as an antioxidant, osmolyte, and calcium modulator such that its intracellular depletion could promote cytotoxicity in diabetes. The relationships of oxidative stress and basal AR gene expression to Na+-taurine cotransporter (TT) gene expression, protein abundance, and TT activity were therefore explored in low AR expressing human retinal pigment epithelial (RPE) 47 cells and RPE 47 cells stably transformed to overexpress AR (RPE 75). Changes in TT gene expression were determined using a 4.6-kb TT promoter-luciferase fusion gene. Compared with RPE 47 cells, in high AR-expressing RPE 75 cells, TT promoter activity was decreased by 46%, which was prevented by an AR inhibitor. TT promoter activity increased up to 900% by prooxidant exposure, which was associated with increased TT peptide abundance and taurine transport. However, induction of TT promoter activity by oxidative stress was attenuated in high AR-expressing cells and partially corrected by AR inhibitor. Finally, exposure of RPE 75 cells to high glucose increased oxidative stress, but down-regulated TT expression. These studies demonstrate for the first time that the TT is regulated by oxidative stress and that overexpression of AR and high glucose impair this response. Abnormal expression of AR may therefore impair antioxidant defense, which may determine tissue susceptibility to chronic diabetic complications. PMID- 16356118 TI - Increased sorbitol pathway activity generates oxidative stress in tissue sites for diabetic complications. AB - Chronic diabetic complications, in particular, nephropathy, peripheral and autonomic neuropathy, "diabetic foot," retinopathy, and cardiovascular disease, remain the major cause of morbidity and mortality in patients with diabetes mellitus. Growing evidence indicates that both increased activity of the sorbitol pathway of glucose metabolism and enhanced oxidative stress are the leading factors in the pathogenesis of diabetic complications. The relation between the two mechanisms remains the area of controversy. One group has reported that increased sorbitol pathway activity has a protective rather than detrimental role in complication-prone tissues because the pathway detoxifies toxic lipid peroxidation products. Others put forward a so-called "unifying hypothesis" suggesting that activation of several major pathways implicated in diabetic complications (e.g., sorbitol pathway) occurs due to increased production of superoxide anion radicals in mitochondria and resulting poly(ADP-ribose) polymerase activation. This review (a) presents findings supporting a key role for the sorbitol pathway in oxidative stress and oxidative stress-initiated downstream mechanisms of diabetic complications, and (b) summarizes experimental evidence against a detoxifying role of the sorbitol pathway, as well as the "unifying concept." PMID- 16356119 TI - Proposed mechanisms for the induction of insulin resistance by oxidative stress. AB - In diabetes (type 1 and type 2), increased flux of free fatty acids and glucose is associated with increased mitochondrial reactive oxygen species (ROS) production and, as a consequence, increased oxidative stress. ROS have been shown to activate various cellular stress-sensitive pathways, which can interfere with cellular signaling pathways. Exposure of different cell lines to micromolar concentrations of hydrogen peroxide leads to the activation of stress kinases such as c-Jun N-terminal kinase, p38, I kappaB kinase, and extracellular receptor kinase 1/2. This activation is accompanied by a down-regulation of the cellular response to insulin, leading to a reduced ability of insulin to promote glucose uptake, and glycogen and protein synthesis. The mechanisms leading to this down regulation in oxidized cells are complicated, involving increased serine/threonine phosphorylation of insulin receptor substrate-1 (IRS1), impaired insulin-stimulated redistribution of IRS1 and phosphatidylinositol-kinase between cytosol and low-density microsomal fraction, followed by a reduced protein kinase B phosphorylation and GLUT4 translocation to the plasma membrane. In addition, prolonged exposure to ROS affects transcription of glucose transporters: whereas the level of GLUT1 is increased, GLUT4 level is reduced. As can be expected, administration of antioxidants such as lipoic acid in oxidized cells, in animal models of diabetes, and in type 2 diabetes shows improved insulin sensitivity. Thus, oxidative stress is presently accepted as a likely causative factor in the development of insulin resistance. PMID- 16356121 TI - Diabetic retinopathy: mitochondrial dysfunction and retinal capillary cell death. AB - Oxidative stress is increased in the retina in diabetes; the levels of oxidatively modified DNA and nitrosylated proteins are elevated, and antioxidant defense enzymes are impaired. The levels of superoxides are elevated in the retina, and the mitochondria become dysfunctional with proapoptotic protein, Bax, translocating from the cytosol into the mitochondria, and cytochrome c leaking out from the mitochondria. This is accompanied by increased retinal capillary cell apoptosis, and the formation of acellular capillaries and pericyte ghosts, the early signs of retinopathy in animal models of diabetic retinopathy. Inhibition of superoxides inhibits glucose -induced mitochondrial dysfunction, activation of caspase-3, and cell death in retinal capillary cells. In animal models, long-term administration of lipoic acid or other antioxidants inhibits the development of diabetic retinopathy via inhibition of accumulation of oxidatively modified DNA and nitrotyrosine and capillary cell apoptosis in the retina. Understanding the role of mitochondria in the development of retinopathy in diabetes should help identify therapies that can neutralize superoxides and inhibit their dysfunction and, ultimately, the development of retinopathy. PMID- 16356120 TI - Role of poly(ADP-ribose) polymerase-1 activation in the pathogenesis of diabetic complications: endothelial dysfunction, as a common underlying theme. AB - Hyperglycemia-induced overproduction of superoxide by mitochondrial electron transport chain triggers several pathways of injury involved in the pathogenesis of diabetic complications [protein kinase C (PKC), hexosamine and polyol pathway fluxes, advanced glycation end product (AGE) formation] by inhibiting glyceraldehyde- 3-phosphate dehydrogenase (GAPDH) activity. Increased oxidative and nitrosative stress activates the nuclear enzyme, poly(ADP-ribose) polymerase 1 (PARP). PARP activation, on the one hand, depletes its substrate, NAD+, slowing the rate of glycolysis, electron transport, and ATP formation. On the other hand, it inhibits GAPDH by poly(ADP-ribosy)lation. These processes result in acute endothelial dysfunction in diabetic blood vessels, which importantly contributes to the development of various diabetic complications. Accordingly, hyperglycemia induced activation of PKC isoforms, hexosaminase pathway flux, and AGE formation is prevented by blocking PARP activity. Furthermore, inhibition of PARP protects against diabetic cardiovascular dysfunction in preclinical models. PARP activation is present in microvasculature of human diabetic subjects. The oxidative/nitrosative stress-PARP pathway leads to diabetes-induced endothelial dysfunction, which may be an important underlying mechanism for the pathogenesis of other diabetic complications (cardiomyopathy, nephropathy, neuropathy, and retinopathy). This review focuses on the role of PARP in diabetic complications and the unique therapeutic potential of PARP inhibition in the prevention or reversal of diabetic complications. PMID- 16356122 TI - Diabetic vascular disease: it's all the RAGE. AB - The major consequence of long-term diabetes is the increased incidence of disease of the vasculature. Of the underlying mechanisms leading to disease, the accumulation of advanced glycation end products (AGEs), resulting from the associated hyperglycemia, is the most convincing. Interaction of AGEs with their receptor, RAGE, activates numerous signaling pathways leading to activation of proinflammatory and procoagulatory genes. Studies in rodent models of macro- and microvascular disease have demonstrated that blockade of RAGE can prevent development of disease. These observations highlight RAGE as a therapeutic target for treatment of diabetic vascular disease. PMID- 16356123 TI - Time-dependent modulation of thioredoxin reductase activity might contribute to sulforaphane-mediated inhibition of NF-kappaB binding to DNA. AB - The chemopreventive agent sulforaphane (SFN) exerts anti-inflammatory activity by thiol-dependent inhibition of nuclear factor kappaB (NF-kappaB) DNA binding. To further analyze the underlying mechanisms, we focused on the thioredoxin/thioredoxin reductase (TrxR) system as a key redox mechanism regulating NF-kappaB DNA binding. Using cultured Raw 264.7 mouse macrophages as a model, 1-chloro-2,4-dinitrobenzene (CDNB), a known inhibitor of TrxR, was identified as an inhibitor of lipopolysaccharide (LPS)-mediated nitric oxide (NO) production and of NF-kappaB DNA binding. CDNB and SFN acted synergistically with respect to inhibition of LPS-induced NO release, and we consequently identified SFN as a novel inhibitor of TrxR enzymatic activity in vitro. Short-term treatment of Raw macrophages with SFN or CDNB resulted in the inhibition of TrxR activity in vivo with half-maximal inhibitory concentration of 25.0 +/- 3.5 microM and 9.4 +/- 3.7 microM, respectively, whereas after a 24-h treatment with 25 microM SFN, TrxR activity was >1.5-fold elevated. In additional experiments, we could exclude that inhibition of trans-activating activity of NF-kappaB contributed to the reduced expression of pro-inflammatory proteins by SFN, based on transient transfection experiments with a (kappaB)(2)- chloramphenicol acetyltransferase construct and a lack of inhibition of protein kinase A activity. These findings further emphasize the importance of redox modulation or thiol reactivity for the regulation of NF-kappaB-dependent transcription by SFN. Antioxid. Redox Signal. 7, 1601-1611. PMID- 16356124 TI - Curcumin inhibits phorbol ester-induced up-regulation of cyclooxygenase-2 and matrix metalloproteinase-9 by blocking ERK1/2 phosphorylation and NF-kappaB transcriptional activity in MCF10A human breast epithelial cells. AB - Elevated levels of cyclooxygenase-2 (COX-2) and matrix metalloproteinases (MMPs) are often observed in various types of cancerous and transformed cells, and hence recognized as potential molecular targets for the chemoprevention. In the present study, we investigated the possible inhibitory effects of curcumin on the expression of COX-2 and MMP-9 induced by the tumor promoter 12-O tetradecanoylphorbol 13-acetate (TPA) in MCF10A human breast epithelial (MCF10A) cells and the underlying mechanisms. Curcumin inhibited the TPA-induced COX-2 expression at both transcriptional and post-transcriptional levels, and reduced the synthesis of prostaglandin E(2), one of the major products of COX-2. Likewise, curcumin attenuated invasiveness and motility of MCF10A cells stimulated with TPA through suppression of MMP expression. Curcumin blocked TPA induced activation of extracellular signal-regulated protein kinase (ERK1/2) and nuclear factor kappaB (NF-kappaB) transcriptional activity. Overexpression of the dominant negative forms of ERK2 abrogated the TPA-induced NF-kappaB transcriptional activity. Treatment of MCF10A cells with U0126, which is a pharmacological inhibitor of ERK1/2, reduced TPA-induced up-regulation of COX-2 and MMP-9. Taken together, these findings suggest that curcumin inhibits the TPA induced up-regulation of COX-2 and MMP-9 by suppressing ERK1/2 phosphorylation and NF-kappaB trans-activation in human breast epithelial cells, which may contribute to its chemopreventive potential. PMID- 16356125 TI - Suppressive effects of mioga ginger and ginger constituents on reactive oxygen and nitrogen species generation, and the expression of inducible pro-inflammatory genes in macrophages. AB - We previously conducted screening tests of the chloroform extracts from a total of 89 species of Japanese plant food items for their suppressive effects on superoxide (O(2) ()) generation through both NADPH oxidase and xanthine oxidase, and reported that mioga ginger (Zingiber mioga Roscoe) indicated the strongest suppressive activities. In this study, the suppressive effects of mioga ginger constituents, aframodial, and galanal B, together with [6]-gingerol and galanolactone occurring in ginger, on free radical generation and inducible proinflammatory gene expressions were investigated. Of these constituents, aframodial (20 microM) exhibited marked suppressive effects on 12-O tetradecanoylphorbol-13-acetate-induced O(2) () generation in HL-60 cells and lipopolysaccharide (LPS)/interferon-gamma-induced nitric oxide (NO) generation in RAW264.7 cells (inhibition rates [IRs]=84.6% and 95.9%, respectively). Aframodial also strongly suppressed the stimulated HL-60 cell-induced mutagenicity in AS52 cells (IR=95.9%). The LPS-induced expression of inducible proinflammatory genes such as inducible NO synthase, interleukin (IL)-1beta, IL-6, and granulocyte macrophage colony-stimulating factor was significantly abolished (IRs=99.1%, 74.6%, 74.0%, and 64.4%, respectively) by aframodial. In addition, degradation of the inhibitor of nuclear factor kappaB was suppressed by this compound (IR=100%), suggesting that the suppression of nuclear factor kappaB activation, at least in part, is involved. Taken together, these results suggest that aframodial has potent antioxidative and anti-inflammatory potentials, and may be a promising candidate in prevention and/or therapy for chronic inflammationassociated carcinogenesis. PMID- 16356126 TI - Chemosensitization and radiosensitization of tumors by plant polyphenols. AB - The treatment of cancer with chemotherapeutic agents and radiation has two major problems: time-dependent development of tumor resistance to therapy (chemoresistance and radioresistance) and nonspecific toxicity toward normal cells. Many plant-derived polyphenols have been studied intently for their potential chemopreventive properties and are pharmacologically safe. These compounds include genistein, curcumin, resveratrol, silymarin, caffeic acid phenethyl ester, flavopiridol, emodin, green tea polyphenols, piperine, oleandrin, ursolic acid, and betulinic acid. Recent research has suggested that these plant polyphenols might be used to sensitize tumor cells to chemotherapeutic agents and radiation therapy by inhibiting pathways that lead to treatment resistance. These agents have also been found to be protective from therapy-associated toxicities. How these polyphenols protect normal cells and sensitize tumor cells to treatment is discussed in this review. PMID- 16356127 TI - Regulation of Nrf2, NF-kappaB, and AP-1 signaling pathways by chemopreventive agents. AB - The inhibition of carcinogenesis by chemopreventive agents has been demonstrated in many tumorigenesis animal models. The chemopreventive mechanisms of those phytochemicals have been investigated extensively, though mostly in in vitro cell culture systems. The cellular signaling cascades mediated by transcription factors, including nuclear factor E2-related factor 2 (Nrf2), nuclear factor kappaB (NF-kappaB), and activator protein-1 (AP-1), have been shown to play pivotal roles in tumor initiation, promotion, and progression processes. Thus, as demonstrated by previous substantive mechanistic studies, they appear to be ideal targets for cancer chemoprevention. In this review, we discuss the current progress and future challenges on our understanding of the molecular mechanisms in cancer chemoprevention by phytochemicals, focusing on the regulation of Nrf2, NF-kappaB, and AP-1 signaling pathways. PMID- 16356128 TI - Molecular mechanism of nrf2 activation by oxidative stress. AB - The capacity of cells to maintain homeostasis during oxidative stress resides in activation or induction of protective enzymes. Nuclear-factor-E2-related factor (Nrf)-2 as a member of bZIP transcription factors is expressed in a variety of tissues. Transcriptional activation of antioxidant genes through an antioxidant response element (ARE) is largely dependent upon Nrf2. The genes that contain a functional ARE include those encoding GSTA1, GSTA2, NAD(P)H:quinone reductase, and gamma-glutamylcysteine synthetase heavy and light subunits that play a role in defense against oxidative stress. Previously, we showed that phosphatidylinositol 3-kinase (PI3-kinase) controls nuclear translocation of Nrf2 in response to oxidative stress, which involves rearrangement of actin microfilaments. Now, we report that PI3-kinase is responsible for the rise of cellular Ca(2+), which is requisite for nuclear translocation of Nrf2. Immunocytochemistry and subcellular fractionation analyses revealed that Nrf2 relocated from the cytoplasm to the plasma membrane prior to its nuclear translocation. We further found that CCAAT/enhancer binding protein-beta (C/EBPbeta), peroxisome proliferatoractivated receptor-gamma (PPARgamma), and retinoid X receptor (RXR) heterodimer serve as the activating transcription factors for the phase II gene induction. Hence, PI3-kinase-mediated Nrf2 activation in combination with activating PPARgamma-RXR and C/EBPbeta contributes to antioxidant phase II enzyme induction via coordinate gene transactivation. PMID- 16356129 TI - Molecular mechanisms involved in enhancing HO-1 expression: de-repression by heme and activation by Nrf2, the "one-two" punch. AB - Heme oxygenase (HO)-1 is a stress response protein, which confers cytoprotection against oxidative injury and provides a vital function in maintaining tissue homeostasis. Molecular mechanisms involved in the inducible transcription of ho-1 occurring in response to numerous and diverse stressful conditions have remained elusive. Since the discovery of E1 and E2, the two upstream enhancers regulating induction of ho-1 transcription in 1989, there have been many studies dealing with molecular mechanisms involved in enhancing HO-1 expression. In this commentary, recent advances in our understanding of the mechanisms involved in the induction of HO-1 expression in mammalian cells are summarized with some supportive results reported by others. Currently available data indicate that activation of ho-1 transcription involves both the heme (native substrate) dependent selective alleviation of repressor and the oxidative stress-dependent activation of transcriptional activator. The stress-released free-heme (HO-1 substrate) from hemoproteins involved in causing oxidative stress itself appears to act as a molecular switch controlling the repressor- activator antagonism on the enhancer sequences of ho-1. Thus, induction of HO-1 appears to operate in a manner like a simple feedback loop. dox Signal. 7, 1674-1687. PMID- 16356130 TI - Molecular basis of heme oxygenase-1 induction: implications for chemoprevention and chemoprotection. AB - Heme oxygenase (HO)-1, involved in the heme degradation process, is an important antioxidant enzyme. The induction of HO-1 gene expression, in response to diverse oxidative stimuli, represents a critical event in adaptive cellular response. Experimental models of various diseases, including acute inflammation, atherosclerosis, degenerative diseases, and carcinogenesis, have demonstrated that the induction of HO-1 can prevent or mitigate the symptoms associated with these ailments. Recent progress in our understanding of cellular signaling networks as critical modulators of gene transcription sheds light on the molecular basis of HO-1 gene expression. A panel of redox-sensitive transcription factors such as activator protein-1, nuclear factor- kappaB, and nuclear factor E2-related factor-2, and some of the upstream kinases have been identified as regulators of HO-1 gene induction. The scope of this review is limited to focus on molecular mechanisms underlying HO-1 expression and the significance of targeted induction of HO-1 as a strategy to achieve chemoprevention and chemoprotection. PMID- 16356131 TI - Redox properties of tea polyphenols and related biological activities. AB - Plant polyphenolic compounds are known to be strong antioxidants. Because oxidative stress is believed to contribute to many acute and chronic diseases, these polyphenols have been postulated to have many beneficial health effects, such as the prevention of cancer and cardiovascular diseases. Indeed, some of these beneficial effects have been demonstrated in animal models and in some, but not all, epidemiological studies. Nevertheless, only some of these activities have been demonstrated to be associated with the antioxidative activities of polyphenols. In studies with cell lines in culture, some of the observed activities may be due to superoxide and hydrogen peroxide produced during the autooxidation of polyphenols. Such pro-oxidation-dependent reactions may not happen in tissues where the oxygen partial pressure is much lower than that in cell culture medium. This review will use the well-studied tea polyphenol, (-) epigallocatechin-3-gallate, as an example to illustrate the redox properties of polyphenols and their influence on signaling pathways related to anti-cancer activities. Existing data suggest, however, that most of the relevant mechanisms of cancer prevention by tea polyphenols are not related to their redox properties, but are due to the direct binding of the polyphenol to target molecules, including the inhibition of selected protein kinases, matrix metalloproteinases, and DNA methyltransferases. PMID- 16356132 TI - Selenium and cancer chemoprevention: hypotheses integrating the actions of selenoproteins and selenium metabolites in epithelial and non-epithelial target cells. AB - The trace element nutrient selenium (Se) discharges its well-known nutritional antioxidant activity through the Se-dependent glutathione peroxidases. It also regulates nuclear factor activities by redox mechanisms through the selenoprotein thioredoxin reductases. Converging data from epidemiological, ecological, and clinical studies have shown that Se can decrease the risk for some types of human cancers, especially those of the prostate, lung, and colon. Mechanistic studies have indicated that the methylselenol metabolite pool has many desirable attributes of chemoprevention, targeting both cancer cells and vascular endothelial cells, whereas the hydrogen selenide pool in excess of selenoprotein synthesis can lead to DNA single strand breaks, which may be mediated by some reactive oxygen species. We propose a new paradigm based on a consideration of the post-initiation biology of avascular early lesion expansion microenvironment, physiochemistry of Se delivery, and the obligatory need for angiogenesis to sustain lesion progression. Our model integrates the roles of selenoproteins and specific Se metabolites to account for cancer risk reduction or enhancement. For future studies, speciation (profiling) methods for Se metabolites and for Se forms in foods and supplements are much needed for hypothesis testing and for the development of mechanism-based Se status markers for cancer prevention. Randomized cancer prevention trials are necessary to test the efficacy of methyl selenium compounds. PMID- 16356133 TI - Evaluation for safety of antioxidant chemopreventive agents. AB - Antioxidants are considered as the most promising chemopreventive agents against various human cancers. However, some antioxidants play paradoxical roles, acting as "double-edged sword." A primary property of effective and acceptable chemopreventive agents should be freedom from toxic effects in healthy population. Miscarriage of the intervention by beta-carotene made us realize the necessity for evaluation of safety before recommending use of antioxidant supplements for chemoprevention. We have evaluated the safety of antioxidants on the basis of reactivity with DNA. Our results revealed that phytic acid, luteolin, and retinoic acid did not cause DNA damage under the experimental condition. Furthermore, phytic acid inhibited the formation of 8-oxo-7,8-dihydro 2'-deoxyguanosine, an indicator of oxidative DNA damage, in cultured cells treated with a H(2)O(2)-generating system. Thus, it is expected that these chemopreventive agents can safely protect humans against cancer. On the other hand, some chemopreventive agents with prooxidant properties (alpha-tocopherol, quercetin, catechins, isothiocyanates, N-acetylcysteine) caused DNA damage via generation of reactive oxygen species in the presence of metal ions and endogenous reductants under some circumstances. Furthermore, other chemopreventive agents (beta-carotene, genistein, daidzein, propyl gallate, curcumin) exerted prooxidant properties after metabolic activation. Therefore, further studies on safety should be required when antioxidants are used for cancer prevention. PMID- 16356134 TI - Thioredoxin and its role in premature newborn biology. AB - Thioredoxin (Trx) is a redox-active protein that has been shown to regulate various cellular processes due to its thiol-disulfide exchange reaction. It has antioxidant properties and also induces the expression of critical antioxidant enzymes such as manganese superoxide dismutase. Trx along with thioredoxin reductase and peroxiredoxins forms a complete system similar to the glutathione system, but with different and divergent functions. This review is a mini-update on key advances in the role of Trx in signal transduction and premature newborn biology. In addition, this mini-update also reviews recently reported prooxidant properties of Trx that relate to anthracycline redox cycling. PMID- 16356135 TI - The role of endothelial cells in the resolution of acute inflammation. AB - Endothelial cells are key regulators of the inflammatory response. Lining blood vessels, they provide in the steady state an antiinflammatory, anticoagulatory surface. However, in the case of injury or infection, endothelial cells control the adhesion and migration of inflammatory cells, as well as the exchange of fluid from the bloodstream into the damaged tissue. Thus, expression of endothelial adhesion molecules, cytokines, and changes in permeability need to be tightly regulated to allow for a controlled inflammatory response. Acute inflammation is characterized by tissue infiltration of neutrophils, followed by monocytes/macrophages. For successful tissue regeneration and healing, the acute inflammatory response needs to be actively shut down, a process called resolution of inflammation. Unsuccessful resolution may lead to excessive tissue damage and ultimately results in chronic, self-promoting inflammation. This review will summarize recent advances in the field of endothelial biology, which point to an active participation of the endothelial barrier in the resolving process. PMID- 16356136 TI - Redox biology of blood revisited: the role of red blood cells in maintaining circulatory reductive capacity. AB - There is an increasing recognition of the role of red blood cells (RBCs) in cell signaling above and beyond its oxygen (O(2))-carrying function. A recent forum published in the December 2004 issue of Antioxidants & Redox Signaling focused on redox biology of blood and the intricate signaling pathways of RBCs or its free components, i.e., hemoglobin, with the vasculature. The forum provided an up-to date source of information on this emerging and exciting area of blood biology and the underlying redox chemistry. In the current short review, we have revisited the topic of redox biology of blood and focused on yet another emerging area of research, which deals with the reductive power of blood and the physiological Redox Signal. PMID- 16356137 TI - The heme oxygenase system: update 2005. PMID- 16356138 TI - Augmentation of sodium butyrate-induced apoptosis by p38 MAP kinase inhibition in rat liver epithelial cells. AB - Sodium butyrate (NaBu) has an inhibitory effect on histone deacetylases (HDACs). The mitogen-activated protein (MAP) kinases, such as extracellular signal regulated kinase 1/2 (ERK1/2) and p38 MAP, kinase are known to be modulated during NaBu-induced apoptosis. In the present study, we showed that low concentrations of NaBu could induce apoptosis synergistically with the inhibition of p38 MAP kinase as proven by using specific p38 MAP kinase inhibitor and dominant negative p38 transfection in a ras-transformed rat liver epithelial cell line (WB-ras). There were no changes in HDAC1, suggesting that NaBu might be able to kill transformed cells bypassing the HDAC inhibitory effect. We further demonstrated that inhibition of p38 MAP kinase potentiated apoptotic cascades, including cleavage of poly(ADP-ribose) polymerase, caspase-3, and decrease in Bcl 2/Bax ratio even at a lower concentration of NaBu. Thus, p38 MAP kinase played inhibitory roles in NaBu-induced apoptosis, and simultaneous modulation of MAP kinases in NaBu treatment could increase the efficiency of the chemotherapeutic effect of NaBu. PMID- 16356139 TI - Differential modulation of MAP kinases by zinc deficiency in IMR-32 cells: role of H(2)O(2). AB - The influence of zinc deficiency on the modulation of the mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinase (ERK1/2), p38, and c-Jun N-terminal kinase (JNK) was studied. Using human IMR-32 cells as a model of neuronal cells, the role of oxidants on MAPKs and activator protein-1 (AP-1) activation in zinc deficiency was investigated, characterizing the participation of these events in the triggering of apoptosis. Relative to controls, cells incubated in media with low zinc concentrations showed increased cell oxidants and hydrogen peroxide (H(2)O(2)) release, increased JNK and p38 activation, high nuclear AP-1-DNA binding activity, and AP-1-dependent gene expression. Catalase addition to the media prevented the increase of cellular oxidants and inhibited JNK, p38, and AP-1 activation. Low levels of ERK1/2 phosphorylation were observed in the zinc-deficient cells in association with a reduction in cell proliferation. Catalase treatment did not prevent the above events nor the increased rate of apoptosis in the zinc-deficient cells. It is first demonstrated that a decrease in cellular zinc triggers H(2)O(2)-independent, as well as H(2)O(2)-dependent effects on MAPKs. Zinc deficiency-induced increases in cellular H(2)O(2) can trigger the activation of JNK and p38, leading to AP-1 activation, events that are not involved in zinc deficiency-induced apoptosis. PMID- 16356141 TI - Low level laser therapy is not low. PMID- 16356142 TI - Intratumor injections of cisplatin and laser thermal therapy for palliative treatment of recurrent cancer. AB - OBJECTIVE: The aim of this work was to report initial findings on the clinical application of intratumor injection of cisplatin in a gel (CDDP/gel) combined with laser-induced thermal therapy (LITT) for cancer treatment in a single patient with advanced stage disease. BACKGROUND DATA: LITT with the neodymium:yttriumaluminum- garnet (Nd:YAG) laser via fiberoptics is a precise, minimally invasive alternative for thermoablation of unresectable or recurrent head and neck neoplasms, but recurrence is often seen at the treatment margins. Combining intratumor chemotherapy with interstitial laser should be most effective using drugs with thermally enhanced toxicity, such as cisplatin. The CDDP/gel therapeutic implant was expected to retain a higher concentration of cisplatin in the tumor margins for improved LITT treatment of the patient presented. METHODS: In this case report, the cisplatin dose was 0.25 mL gel/cm(3) tumor volume (20 mg of CDDP) followed by LITT (Nd:YAG laser, 50 W, PD = 2,200 J/cm(2)) after the chemotherapy session. RESULTS: The patient responded with local tumor eradication, and no signs of systemic toxicity were observed related to this therapy. However, the patient developed progressive metastatic disease in the lungs and died 2.5 months later. CONCLUSIONS: This is a report of a patient with an accessible solid tumor who was treated with intratumor injection of CDDP/gel followed by LITT, which proved to be feasible. Based on preclinical evidence obtained at UCLA and the results of this study, we are encouraged to continue our refinement of LITT combined with chemotherapy for cancer treatment. PMID- 16356143 TI - Endoluminal phototherapy for prevention of restenosis: preliminary results at 6 month follow-up. AB - OBJECTIVE: The objective of this observational study was to investigate safety and efficacy of laser phototherapy (LPh) in prevention of restenosis after percutaneous coronary intervention (PCI). BACKGROUND DATA: Laser irradiation is known to cause a limitation of the local inflammatory cascade and a stimulation of proliferation of specific cells. Based on the results of previous experiments proving the beneficial effects of laser light on the activity of vascular and inflammatory cells, we attempted to use these properties to prevent restenosis. METHODS: Laser phototherapy was performed in 41 patients after stent implantation or balloon angioplasty. Illumination power of 100 mW and energy dose equal to 9 J/cm(2) was used. Patients were monitored for major adverse cardiac events (MACE) after 30 days and 6 months. At 6 months, angiography as a control was performed to assess the influence of LPh on restenosis rate. RESULTS: Angiographic follow up (n = 30) revealed restenosis in 9% and 25% of patients after stent implantation and balloon angioplasty, respectively. The MACE rate was 4.5% and 12.5% in stent and balloon-treated patients, respectively. CONCLUSIONS: Laser phototherapy gives very promising results in restenosis prevention, especially after stent implantation. The treatment method is safe, with a low rate of MACE in follow-up. PMID- 16356144 TI - Micro-tensile bond strength between a resin cement and an aluminous ceramic treated with Nd:YAG laser, Rocatec System, or aluminum oxide sandblasting. AB - OBJECTIVE: The aim of this study was to evaluate the effect of different surface treatments on bond strength between In-Ceram Alumina (VITA) and Panavia Fluoro Cement (Kuraray), and to verify the association between bond strength values and the percentage of remaining cement on the ceramic surface. BACKGROUND DATA: Although several conventional surface treatments have been used on aluminous ceramic, no study investigated the effect of the Nd:YAG laser. METHODS: Nine In Ceram Alumina blocks (10 x 10 x 6 mm) received three surface treatments: group 1, Al(2)O(3) sandblasting + silane; group 2, Al(2)O(3) sandblasting + Rocatec Plus + silane; and group 3, Al(2)O(3) sandblasting + Nd:YAG Laser + silane. Resin cement was applied and light cured, followed by the application of composite. Blocks were sectioned to obtain sticks with a cross-sectional area of 1.0 mm(2). Microtensile bond strength (microTBS) was recorded in a universal testing machine at a crosshead speed of 0.5 mm/min. Scanning electron microscopy images of the fractured areas were used to calculate the amount of remaining cement on the ceramic substrate. RESULTS: All treatments yielded statistically different microTBS values (MPa): group 1, 14.38(A); group 2, 20.09(B); and group 3, 23.21(C) (ANOVAand Tukey's test, alpha = 0.05). Pearson's correlation coefficient for microTBS x percentage of remaining cement was r = 0.92 (p < 0.05). CONCLUSIONS: The Nd:YAG laser was the most effective surface treatment, followed by Rocatec and Al(2)O(3) sandblasting. A strong positive association was found between bond strength values and percentage of remaining cement. PMID- 16356145 TI - Influence of visible light and ultraviolet irradiation on motility and fertility of mammalian and fish sperm. AB - OBJECTIVE: The effects of visible light irradiation on sperm motility, fertility, and reactive oxygen species (ROS) formation were investigated and compared in ram and fish (tilapia). BACKGROUND DATA: Low-energy visible light has previously been found to modulate various processes in different biological systems. In the literature, it is accepted that the first step following visible light irradiation is the formation of ROS by endogenous cellular photosensitizers. METHODS: Sperm of ram and tilapia were irradiated with various light sources (400 800 nm white light, 660 nm red light, 360 nm blue light, 294 nm UV), and their motility and fertility rates were measured. The amount of ROS generated by irradiation was estimated using electron paramagnetic resonance (EPR) technique. RESULTS: Sperm taken from tilapia showed higher motility and fertility following red and white light irradiation. In contrast, the motility and fertility of ram sperm were slightly increased only by red light. A negative effect on motility and fertility of sperm of both species was obtained following irradiation with UV and blue light. The amount of ROS produced in irradiated tilapia sperm was much higher than that of ram sperm. CONCLUSIONS: The results show that different wavelengths differentially affect tilapia and ram sperm motility and fertilization. The difference in response to the various light sources might be explained by the different amounts of ROS formation by ram and tilapia, which are in agreement with the physiology of fertilization appropriate to each of these species. Based on these results, it is suggested that in vitro fertilization in mammals should be performed in darkness or at least under red light. PMID- 16356146 TI - Low-level laser therapy improves early healing of medial collateral ligament injuries in rats. AB - OBJECTIVE: This study sought to investigate whether or not low-level laser therapy (LLLT) with a helium-neon laser increased biomechanical parameters of transected medial collateral ligament (MCL) in rats. BACKGROUND DATA: It has been reported that LLLT can enhance tendon healing. METHODS: Thirty rats received surgical transection to their right MCL, and five were assigned as the control group. After surgery, the rats were divided into three groups: group 1 (n = 10) received LLLT with 0.01 J/cm(2) energy density per day, group 2 (n = 10) received LLLT with 1.2 J/cm(2) energy density per day, and group 3 (sham = exposed group; n = 10) received daily placebo laser with shut-down laser equipment, while the control group received neither surgery nor LLLT. Biomechanical tests were performed at 12 and 21 days after surgery. The data were analyzed by one-way analysis of variance. RESULTS: The ultimate tensile strength (UTS) of group 2 on day 12 was significantly higher than that of groups 1 and 3. Furthermore, the UTS and energy absorption of the control (uninjured) group were significantly higher than those of the other groups. CONCLUSIONS: LLLT with a helium-neon laser is effective for the early improvement of the ultimate tensile strength of medial collateral ligament injuries. PMID- 16356147 TI - XeCl excimer laser ablation of rabbit tibia bone: morphology of the irradiated site and self-limiting effect. AB - OBJECTIVE: The purpose of this work is to investigate the morphological changes of the rabbit tibia bone submitted to osteotomy with XeCl excimer laser. BACKGROUND DATA: Laser ablation of tissue is a relevant clinical application of high-power lasers. Due to the variety of laser wavelengths and properties of biological targets, the subject has not yet been completely explored. METHODS: In this study, four tibias were irradiated with six different durations. The applied energy density was 6.7 J/cm(2) per pulse at a 4-Hz repetition rate. RESULTS: The 24 samples had similar and well-defined craters that were free of carbonization. The ablation rates per pulse, at 60-120 seconds, were 3.8 and 1.9 microm at the compact bone and the marrow tissue, respectively. The ablation process ceased after 120 sec, with a 1.6-mm crater depth. CONCLUSIONS: In the present study, the laser used showed a self-limiting characteristic, which is a phenomenon that can be of great value in preserving important structures close to the working area, giving a margin of safety to the surgeon, in case of over-irradiation. This self limiting effect is attributed to liquid filling the cavity by the bleeding and irrigation process. PMID- 16356148 TI - Evaluation of low intensity laser effects on the thyroid gland of male mice. AB - OBJECTIVE: The purpose of this study was to assess whether there were alterations in the thyroid hormone plasma levels under infrared laser irradiation, in the thyroid gland region. BACKGROUND DATA: Studies have demonstrated that infrared laser can cause alterations in thyroid glands. METHODS: Sixty-five albino male mice were used and assigned to five groups (n = 13), with differences in the times that they were sacrificed. Irradiation procedures consisted of an infrared diode laser emitting at 780 nm, at 4 J/cm(2) energy density, in contact mode, point manner. Blood was collected before irradiation (group 1), and then at 24 h (group 2), 48 h (group 3) and 72 h (group 4), and 1 week (group 5) after the third irradiation. The collected material was used for clinical analysis to evaluate the T(3) (triiodothyronine) and T(4) (thyroxin) hormones. Five animals were used for light microscopy analysis. RESULTS: A statistically significant hormonal level alteration between the first day and 7 days after the last irradiation was found. CONCLUSIONS: It was concluded that low-level laser therapy (LLLT) of the thyroid gland may affect the level of thyroidal hormones. PMID- 16356149 TI - Helium-neon and nitrogen laser irradiation accelerates the phagocytic activity of human monocytes. AB - OBJECTIVE: Intracellular survival of mycobacteria within monocytes is a crucial stage in the pathogenesis of tuberculosis. The aim was to check intracellular survival of Mycobacterium fortuitum within the human monocytes exposed to He-Ne and nitrogen laser irradiation. BACKGROUND DATA: Tuberculosis remains one of the most important infectious diseases for developing countries. Low-level laser therapy (LLLT) has been tried to treat tubercular cavitory lung disease with encouraging results. The in vitro photobiological effect of low level laser radiation on the intracellular mycobacteria needs to be evaluated before we could go for large clinical trials. METHODS: The aliquots of human monocytes from peripheral blood of healthy volunteers and tuberculosis cases were exposed to He Ne or nitrogen laser beam. The non-irradiated monocytes from the same source served as controls. The monocytes were then challenged with M. fortuitum, and surviving mycobacteria within monocytes were subjected to viable counts. RESULTS: Enhanced killing of mycobacterial cells was seen among monocytes exposed to He-Ne and nitrogen laser irradiation. CONCLUSIONS: He-Ne and nitrogen laser irradiation activates the monocytes to increase intracellular killing of mycobacteria. PMID- 16356150 TI - Effects of diode laser irradiation on smear layer removal from root canal walls and apical leakage after obturation. AB - OBJECTIVE: The objective of this study was to investigate the rise in temperature in root surfaces during and immediately after diode laser irradiation, to observe morphological changes of root canal wall after irradiation, and to evaluate the apical leakage after irradiation and obturation in vitro. BACKGROUND DATA: There have been very few reports on root canal treatment by 980-nm wavelength diode laser. METHODS: Sixty-six extracted human single-rooted teeth were instrumented up to size 60 K-file, and then randomly divided into three groups of 22 teeth each. Groups 1 and 2 were irradiated with a diode laser at 5 W for 7 sec using fibers of diameters 550 and 365 microm, respectively. Group 3 was not irradiated, and served as a control. The rise in temperature on root surfaces of the teeth in groups 1 and 2 were measured by thermography. Six teeth in each group were bisected longitudinally and observed morphologically. Other teeth were obturated and immersed in rhodamine B solution, and the degree of apical leakage was evaluated longitudinally and transversally. RESULTS: A maximum temperature rise of 8.1( degrees )C was recorded in group 1. The smear layer in the laser-treated groups was evaporated and removed, resulting in clean root canal walls, which was significantly superior to the control group (p < 0.05). After obturation, the laser-treated groups showed significantly less apical leakage than the control group (p < 0.05). CONCLUSIONS: These results indicate that the diode laser is useful for removing smear layer and debris from root canal walls, and reducing apical leakage after obturation in vitro, and suggest that it would be useful for root canal treatment in clinic. PMID- 16356151 TI - In vivo study of the Nd:YAP laser in persistent periapical lesion. AB - OBJECTIVE: The aim of this study was to evaluate the therapeutic effect of ND:YAP laser in persistent lesions through signs and symptoms such as the presence of fistulas and pain, respectively. BACKGROUND DATA: Periapical lesions with bacterial invasion, giving origin to bacterial infections, appear many times during the endodontic treatment, which aims at preparing and disinfecting the root canal system, in canals with mortified pulp. The endodontic treatment aim at eliminating infections and reinfection prevention; however, sometimes re treatment is necessary, and it can be complemented with paraendodontic surgery due to reinfection, which can also persist. METHODS: Our study group consisted of six clinical cases (endodontic treatment), with the introduction of a 300-microm optic fiber through the fistula channel, where three applications were carried out with an interval of 15 sec, each with a discharge of 300 mJ of energy and frequency of 30 Hz. RESULTS: After a 7-day interval, the signs and symptoms were absent, even after a follow-up of 18 months. It was also observed, through a microbiological study, the decrease of the local microbiota. CONCLUSIONS: Our results supported the use of Nd:YAP laser, in part since it created an unfavorable environment for the continuing development of microorganisms. PMID- 16356152 TI - The effect of laser therapy on the proliferation of oral KB carcinoma cells: an in vitro study. AB - OBJECTIVE: The aim of this study was to assess the proliferative effect of carcinoma cells, strain KB, submitted to laser therapy with wavelengths of lambda685nm (31 mW; O; 0.38 cm(2), 4 J/cm(2)) or lambda830nm (34.5 mW; O; 0.38 cm(2), 4 J/cm(2)). BACKGROUND DATA: It is known that the interaction of laser light with living tissues may lead to different results depending upon several factors such as wavelength, dose, potency, and optical properties of the tissue as well as on the condition being treated. The response to the use of laser light may be of stimulation or inhibition. One successful model used to study the effects of laser light on living tissues is the in vitro use of different lineages of cells in culture. METHODS: Cellular viability was assessed using MTT spectroscopy immediately, and 6, 12, 24, and 72 h after treatment. The irradiations were carried out twice, at 24 h after cell seeding and at 48 h after the first irradiation. The dose of 4 J/cm(2) was given by a lambda685 nm (31 mW, Phi 0.8 cm(2)) or lambda830 nm (34.5 mW, Phi 0.8 cm(2)) diode lasers. RESULTS: The results demonstrated that the time influenced significantly both control (p = 0.01) and both cultures irradiated with lambda685-nm laser (p = 0.01) or lambda830-nm laser (p = 0.09). The influence of the treatment (laser therapy) was also significant when comparing the results observed in irradiated groups and the control (p = 0.01). The influence of the wavelength in the final result, in other words, in the cellular viability of cultures irradiated with the two wavelengths was also significant (p = 0.01). CONCLUSIONS: It is concluded that laser therapy had a positive biomodulatory effect on the proliferation of KB cells and that this was influenced by the wavelength. PMID- 16356153 TI - Morphologic evaluation of the radicular dentine irradiated with Nd:YAG laser under different parameters and angles of incidence. AB - OBJECTIVE: The aim of this research was to evaluate, under the scanning electron microscope, the role of different parameters and angles of incidence of the Nd:YAG laser on the morphology of the radicular dentine. BACKGROUND DATA: Irradiation of dentine with Nd:YAG laser removes smear layer and promotes its fusion and recrystallization, thus decreasing permeability. METHODS: Forty-eight human maxillary canines were randomly divided in four groups of 12 teeth each, according to the following parameters of irradiation: 1 Wand 10 Hz; 1 Wand 20 Hz; 3 Wand 10 Hz; 3 Wand 20 Hz. Two teeth of each group were split longitudinally and the laser was applied perpendicularly to the root canal. Specimens were observed under the SEM, and the morphological alterations in dentine were recorded. RESULTS: The increase of power and frequency of the laser produced more structures that characterize the irradiation of dentine by Nd:YAG laser, such as globular formations, melting and glazing, and ebullitions. Such structures were especially frequent when 3 Wand 20 pps were used as parameters. The removal of the smear layer produced root canals with more dentinal tubules visible under the SEM. CONCLUSIONS: The increase in power and frequency of the Nd:YAG laser irradiation produced more visible morphological alterations than the lower settings. The removal of the smear layer increased the number of visible dentinal tubule openings. PMID- 16356154 TI - Sacroiliac joint debridement: a novel technique for the treatment of sacroiliac joint pain. AB - OBJECTIVE: This study was a retrospective analysis of 38 patients who underwent sacroiliac joint debridement (SJD) as a treatment for confirmed sacroiliac joint (SI joint) pain. BACKGROUND DATA: This is a new, unpublicized, minimally invasive (we define minimally invasive as a surgery with an incision of less than 1 inch) surgical technique. There are no prior studies on this surgery, but the surgery is compared to SI joint fusion surgery, which offers a success rate of 50-70% in larger studies. METHODS: Thirty-eight patients with confirmed SI joint pain via a preoperative modified SI joint injection underwent SJD. These patients were followed up at 12-month intervals to determine their degree of pain relief from this surgery. RESULTS: Of the 38 patients, 61% of these patients had 50-100% reductions of their VAS and 53% had >75% improvement for >2 years. No complications were noted. Histology sections of the removed tissues revealed a non-inflammatory degenerative musculo-tendinous tissue similar to a chronic tendonitis. CONCLUSIONS: SJD is a reasonable treatment option for SI joint pain, which has a low complication rate and a success rate similar to SI joint fusion. Sacroiliac joint pain may be related to a degenerative musculo-tendinous condition of the surface of the joint on the iliac side. PMID- 16356155 TI - Laser literature watch. PMID- 16356162 TI - Bile acid profiles by capillary electrophoresis in intrahepatic cholestasis of pregnancy. AB - ICP (intrahepatic cholestasis of pregnancy) is characterized by pruritus and biochemical cholestasis, including raised SBAs (serum bile acids) and, usually, elevated aminotransferases levels. However, AHP (asymptomatic hypercholanaemia of pregnancy) is defined as the presence of total SBA levels above the cut-off value (11 microM) in healthy pregnant women, thus elevation of total SBAs do not necessarily reflect an ICP condition. The aim of the present study was to describe clinical, obstetric, perinatal and biochemical findings, as well as the SBA profile, in pregnant women studied in the third trimester of pregnancy in order to define characteristic patterns of individual bile acids that enable women with ICP to be distinguished from AHP and healthy pregnancies. Free and conjugated ursodeoxycholic (UDCA), cholic (CA), lithocholic (LCA), deoxycholic (DCA) and chenodeoxycholic (CDCA) acids were evaluated by CE (capillary electrophoresis) in 41 patients (15 of them simultaneously by HPLC), in 30 healthy pregnant women and in 10 non-pregnant women. A highly significant correlation between CE and HPLC for total SBAs (r=0.990) and for individual SBAs was found. Normal pregnant women had higher total SBA levels than non-pregnant women (due to an increase in taurine-conjugated dihydroxy SBAs). Women with ICP had higher levels of total SBAs, the free/conjugated ratio, LCA, CA, CDCA and DCA than normal pregnant women. Newborns from women with ICP had lower birth weight and gestational age. Women with AHP had higher levels of conjugated dihydroxy SBAs than normocholanaemic patients, without any evidence of a clinical difference. In conclusion, the present study has shown a clear difference in SBA profiles between ICP and normal pregnancies (including AHP), involving a shift towards a characteristic hydrophobic composition in women with ICP. PMID- 16356163 TI - A comparison between active- and reactive-hyperaemia-induced brachial artery vasodilation. AB - The measurement of brachial artery vasodilation in response to a hyperaemic stimulus has been used extensively to assess changes in endothelial function. However, whether or not similar changes occur in response to an active hyperaemic stimulus is unknown. The purpose of the present study was to compare brachial artery vasodilation in response to an active compared with a reactive hyperaemic stimulus following a known perturbation of endothelial function. Eight apparently healthy adults were assigned to four treatment conditions in a counter-balanced design: (i) low-fat meal with active hyperaemic stimulus (LFM-A), (ii) high-fat meal with active hyperaemic stimulus (HFM-A), (iii) low-fat meal with reactive hyperaemic stimulus (LFM-R), and (iv) high-fat meal with reactive hyperaemic stimulus (HFM-R). Meals were ingested at 08:00 hours on each treatment day. Brachial artery vasodilation was assessed via ultrasound 4 h after ingestion of each meal. The active hyperaemic stimulus was induced by 5 min of rhythmic handgrip exercise, whereas reactive hyperaemia was induced by 5 min of forearm occlusion. Brachial artery vasodilation was expressed as the percentage change in diameter from baseline to post-active/reactive hyperaemia. Using a 2x2 repeated measures ANOVA, a significant stimulusxmeal interaction (P=0.025) was found. Simple main effects revealed no difference (P=0.541) in brachial artery vasodilation between LFM-A (5.75+/-1.64%) and HFM-A (6.39+/-1.45%); however, a significant decrease (P=0.014) in brachial artery vasodilation was found in the HFM-R (4.29+/-1.64%) compared with the LFM-R (7.18+/-1.13%) treatment. In conclusion, the measurement of brachial artery vasodilation in response to active hyperaemia did not detect a change in endothelial function following a single perturbation meal, whereas reactive hyperaemia did. PMID- 16356164 TI - Haemoglobin and flow-mediated vasodilation. AB - A low [Hb] (Hb concentration) is out-balanced by peripheral vasodilation via mechanisms that are incompletely understood. Peripheral vasodilation is influenced by NO (nitric oxide) released from vascular endothelium in response to increased vessel wall shear stress, and absorption by Hb is the main mechanism by which the bioactivity of NO is disarmed. Thus we propose that graded NO absorption is the mechanism through which a low [Hb] is related to peripheral vasodilation. In the present study, we examined the relationship between [Hb] and FMD (flow-mediated vasodilation; 5 min of cuff ischaemia) of the radial and brachial arteries in 33 normal subjects and in 13 patients with Type II diabetes, known to have impaired NO-mediated vasodilation. The smaller radial artery provided the more sensitive test, as it had a 2-fold larger FMD than the brachial artery (22+/-18% compared with 9+/-18% respectively, in normal subjects; means+/ S.D., P<0.05). FMD of the radial artery had a negative correlation with [Hb] (r(2)=-0.66, P<0.05; n=27). In subjects with [Hb] below and above the median of 14.1 g/dl, the radial artery FMD was 30+/-22% compared with 13+/-12% respectively (P<0.05). In diabetic patients, FMD was lower and a co-variation with [Hb] could not be established. Thus, in normal subjects, NO-mediated endothelium-related vasodilation at least partly out-balanced the 'added burden' of a low [Hb] during post-ischaemic reperfusion. PMID- 16356165 TI - Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 (GRK2) in HEK-293beta2 cells and cardiac myocytes. AB - Membrane-recruitment of GRK2 (G-protein receptor kinase 2) provides a fundamental step in the desensitization process controlling GPCRs (G-protein-coupled receptors), such as the beta2AR (beta2-adrenergic receptor). In the present paper, we show that challenge of HEK-293beta2 [human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP (green fluorescent protein)] cells with the beta-adrenoceptor agonist, isoprenaline, causes GRK2 to become phosphorylated by PKA (cAMP-dependent protein kinase). This action is facilitated when cAMP-specific PDE4 (phosphodiesterase-4) activity is selectively inactivated, either chemically with rolipram or by siRNA (small interfering RNA) mediated knockdown of PDE4B and PDE4D. PDE4-selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of GRK2, phosphorylation of the beta2AR by GRK2, membrane translocation of beta-arrestin and internalization of beta2ARs. PDE4-selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of GRK2 in cardiac myocytes. In the absence of isoprenaline, rolipram-induced inhibition of PDE4 activity in HEK-293beta2 cells acts to stimulate PKA phosphorylation of GRK2, with consequential effects on GRK2 membrane recruitment and GRK2-mediated phosphorylation of the beta2AR. We propose that a key role for PDE4 enzymes is: (i) to gate the action of PKA on GRK2, influencing the rate of GRK2 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge, and (ii) to protect GRK2 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of cAMP. PMID- 16356166 TI - Kinetic analysis using low-molecular mass xyloglucan oligosaccharides defines the catalytic mechanism of a Populus xyloglucan endotransglycosylase. AB - Plant XETs [XG (xyloglucan) endotransglycosylases] catalyse the transglycosylation from a XG donor to a XG or low-molecular-mass XG fragment as the acceptor, and are thought to be important enzymes in the formation and remodelling of the cellulose-XG three-dimensional network in the primary plant cell wall. Current methods to assay XET activity use the XG polysaccharide as the donor substrate, and present limitations for kinetic and mechanistic studies of XET action due to the polymeric and polydisperse nature of the substrate. A novel activity assay based on HPCE (high performance capillary electrophoresis), in conjunction with a defined low-molecular-mass XGO {XG oligosaccharide; (XXXGXXXG, where G=Glcbeta1,4- and X=[Xylalpha1,6]Glcbeta1,4-)} as the glycosyl donor and a heptasaccharide derivatized with ANTS [8-aminonaphthalene-1,3,6-trisulphonic acid; (XXXG-ANTS)] as the acceptor substrate was developed and validated. The recombinant enzyme PttXET16A from Populus tremula x tremuloides (hybrid aspen) was characterized using the donor/acceptor pair indicated above, for which preparative scale syntheses have been optimized. The low-molecular-mass donor underwent a single transglycosylation reaction to the acceptor substrate under initial-rate conditions, with a pH optimum at 5.0 and maximal activity between 30 and 40 degrees C. Kinetic data are best explained by a ping-pong bi-bi mechanism with substrate inhibition by both donor and acceptor. This is the first assay for XETs using a donor substrate other than polymeric XG, enabling quantitative kinetic analysis of different XGO donors for specificity, and subsite mapping studies of XET enzymes. PMID- 16356167 TI - Lipid phosphate phosphatase-1 regulates lysophosphatidic acid- and platelet derived-growth-factor-induced cell migration. AB - LPPs (lipid phosphate phosphatases) are members of a family of enzymes that catalyse the dephosphorylation of lipid phosphates. The only known form of regulation of this family of enzymes is via de novo expression of LPP isoforms in response to growth factors. In this respect, we evaluated the effect of moderate increases in the expression of recombinant LPP1 on signal transduction by both G protein-coupled receptors and receptor tyrosine kinases. We present evidence for a novel role of LPP1 in reducing PDGF (platelet-derived growth factor)- and lysophosphatidic acid-induced migration of embryonic fibroblasts. We demonstrate that the overexpression of LPP1 inhibits cell migration by reducing the PDGF induced activation of p42/p44 MAPK (mitogen-activated protein kinase). This appears to occur via a mechanism that involves the LPP1-induced down-regulation of typical PKC (protein kinase C) isoform(s), which are normally required for PDGF-induced activation of p42/p44 MAPK and migration. In this regard, DAG (diacylglycerol) levels are high and sustained in cells overexpressing LPP1, suggesting a dynamic interconversion of phosphatidic acid into DAG by LPP1. This may account for the effects of LPP1 on cell migration, as sustained DAG is known to down-regulate PKC isoforms in cells. Therefore the physiological changes in the expression levels of LPP1 might represent a heterologous desensitization mechanism for attenuating PKC-mediated signalling and regulation of cell migration. PMID- 16356168 TI - Gene expression profiling of mice with genetically modified muscle glycogen content. AB - Glycogen, a branched polymer of glucose, forms an energy re-serve in numerous organisms. In mammals, the two largest glyco-gen stores are in skeletal muscle and liver, which express tissue-specific glycogen synthase isoforms. MGSKO mice, in which mGys1 (mouse glycogen synthase) is disrupted, are devoid of muscle glycogen [Pederson, Chen, Schroeder, Shou, DePaoli-Roach and Roach (2004) Mol. Cell. Biol. 24, 7179-7187]. The GSL30 mouse line hyper-accumulates glycogen in muscle [Manchester, Skurat, Roach, Hauschka and Lawrence (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 10707-10711]. We performed a microarray analysis of mRNA from the anterior tibialis, medial gastrocnemius and liver of MGSKO mice, and from the gastroc-nemius of GSL30 mice. In MGSKO mice, transcripts of 79 genes varied in their expression in the same direction in both the anterior tibialis and gastrocnemius. These included several genes encoding proteins proximally involved in glycogen metabolism. The Ppp1r1a [protein phosphatase 1 regulatory (inhibitor) sub-unit 1A] gene underwent the greatest amount of downregulation. In muscle, the downregulation of Pfkfb1 and Pfkfb3, encoding isoforms of 6-phosphofructo-2 kinase/fructose-2,6-bisphospha-tase, is consistent with decreased glycolysis. Pathways for branched-chain amino acid, and ketone body utilization appear to be downregulated, as is the capacity to form the gluconeogenic precursors alanine, lactate and glutamine. Expression changes among several members of the Wnt signalling pathway were identified, suggesting an as yet unexplained role in glycogen meta-bolism. In liver, the upregulation of Pfkfb1 and Pfkfb3 expression is consistent with increased glycolysis, perhaps as an adaptation to altered muscle metabolism. By comparing changes in muscle expression between MGSKO and GSL30 mice, we found a subset of 44 genes, the expression of which varied as a function of muscle glycogen content. These genes are candidates for regulation by glycogen levels. Particularly interesting is the observation that 11 of these genes encode cardiac or slow-twitch isoforms of muscle contractile proteins, and are upregulated in muscle that has a greater oxidative capacity in MGSKO mice. PMID- 16356169 TI - Lack of ceramide generation and altered sphingolipid composition are associated with drug resistance in human ovarian carcinoma cells. AB - PTX (Paclitaxel) is an antimitotic agent used in the treatment of a number of major solid tumours, particularly in breast and ovarian cancer. This study was undertaken to gain insight into the molecular alterations producing PTX resistance in ovarian cancer. PTX treatment is able to induce apoptosis in the human ovarian carcinoma cell line, CABA I. PTX-induced apoptosis in CABA I cells was accompanied by an increase in the cellular Cer (ceramide) levels and a decrease in the sphingomyelin levels, due to the activation of sphingomyelinases. The inhibition of acid sphingomyelinase decreased PTX-induced apoptosis. Under the same experimental conditions, PTX had no effect on Cer and sphingomyelin levels in the stable PTX-resistant ovarian carcinoma cell line, CABA-PTX.The acquisition of the PTX-resistant phenotype is accompanied by unique alterations in the complex sphingolipid pattern found on lipid extraction. In the drug resistant cell line, the levels of sphingomyelin and neutral glycosphingolipids were unchanged compared with the drug-sensitive cell line. The ganglioside pattern in CABA I cells is more complex compared with that of CABA-PTX cells. Specifically, we found that the total ganglioside content in CABA-PTX cells was approximately half of that in CABA I cells, and GM3 ganglioside content was remarkably higher in the drug-resistant cell line. Taken together our findings indicate that: i) Cer generated by acid sphingomyelinase is involved in PTX induced apoptosis in ovarian carcinoma cells, and PTX-resistant cells are characterized by their lack of increased Cer upon drug treatment, ii) PTX resistance might be correlated with an alteration in metabolic Cer patterns specifically affecting cellular ganglioside composition. PMID- 16356170 TI - Rapid screening for glucose-6-phosphate dehydrogenase deficiency and haemoglobin polymorphisms in Africa by a simple high-throughput SSOP-ELISA method. AB - BACKGROUND: Mutations in the haemoglobin beta-globin (HbB) and glucose-6 phosphate dehydrogenase (G6PD) genes cause widespread human genetic disorders such as sickle cell diseases and G6PD deficiency. In sub-Saharan Africa, a few predominant polymorphic variants of each gene account for a majority of these deficiencies. Examining at a larger scale the clinical importance of these independent genetic disorders, their possible association with malaria pathogenesis and innate resistance, and their relevance for antimalarial drug treatment, would be easier if an accurate screening method with limited costs was available. METHODS: A simple and rapid technique was developed to detect the most prominent single nucleotide polymorphisms (SNPs) in the HbB and G6PD genes. The method is able to detect the different haemoglobin polymorphisms A, S, C and E, as well as G6PD polymorphisms B, A and A- based on PCR-amplification followed by a hybridization step using sequence-specific oligonucleotide probes (SSOPs) specific for the SNP variants and quantified by ELISA. RESULTS: The SSOP-ELISA method was found to be specific, and compared well to the commonly used PCR-RFLP technique. Identical results were obtained in 98% (haemoglobin) and 95% (G6PD) of the tested 90 field samples from a high-transmission area in Tanzania, which were used to validate the new technique. CONCLUSION: The simplicity and accuracy of the new methodology makes it suitable for application in settings where resources are limited. It would serve as a valuable tool for research purposes by monitoring genotype frequencies in relation to disease epidemiology. PMID- 16356171 TI - Fast-track applications: the potential for direct delivery of proteins and nucleic acids to plant cells for the discovery of gene function. AB - In animal systems, several methods exist for the direct delivery of nucleic acids and proteins into cells for functional analysis. Until recently, these methods have not been applied to plant systems. Now, however, several preliminary reports suggest that both nucleic acids and proteins can also be delivered into plant cells by very simple, direct application. This promises to open the way for high throughput screening for gene function in a range of plant species. PMID- 16356172 TI - Analysis of concordance of different haplotype block partitioning algorithms. AB - BACKGROUND: Different classes of haplotype block algorithms exist and the ideal dataset to assess their performance would be to comprehensively re-sequence a large genomic region in a large population. Such data sets are expensive to collect. Alternatively, we performed coalescent simulations to generate haplotypes with a high marker density and compared block partitioning results from diversity based, LD based, and information theoretic algorithms under different values of SNP density and allele frequency. RESULTS: We simulated 1000 haplotypes using the standard coalescent for three world populations--European, African American, and East Asian--and applied three classes of block partitioning algorithms--diversity based, LD based, and information theoretic. We assessed algorithm differences in number, size, and coverage of blocks inferred under different conditions of SNP density, allele frequency, and sample size. Each algorithm inferred blocks differing in number, size, and coverage under different density and allele frequency conditions. Different partitions had few if any matching block boundaries. However they still overlapped and a high percentage of total chromosomal region was common to all methods. This percentage was generally higher with a higher density of SNPs and when rarer markers were included. CONCLUSION: A gold standard definition of a haplotype block is difficult to achieve, but collecting haplotypes covered with a high density of SNPs, partitioning them with a variety of block algorithms, and identifying regions common to all methods may be the best way to identify genomic regions that harbor SNP variants that cause disease. PMID- 16356173 TI - The effect of cartilage and bone density of mushroom-shaped, photooxidized, osteochondral transplants: an experimental study on graft performance in sheep using transplants originating from different species. AB - BACKGROUND: Differences in overall performance of osteochondral photooxidized grafts were studied in accordance of their species origin and a new, more rigorous cleansing procedure using alcohol during preparation. METHODS: Photooxidized mushroom-shaped grafts of bovine, ovine, human and equine origin were implanted in the femoral condyles of 32 sheep (condyles: n = 64). No viable chondrocytes were present at the time of implantation. Grafts were evaluated at 6 months using plastic embedded sections of non-decalcified bone and cartilage specimens. Graft incorporation, the formation of cyst-like lesions at the base of the cartilage junction as well as cartilage morphology was studied qualitatively, semi-quantitatively using a score system and quantitatively by performing histomorphometrical measurements of percentage of bone and fibrous tissue of the original defects. For statistical analysis a factorial analysis of variance (ANOVA- test) was applied. RESULTS: Differences of graft performance were found according to species origin and cleansing process during graft preparation. According to the score system cartilage surface integrity was best for equine grafts, as well as dislocation or mechanical stability. The equine grafts showed the highest percentage for bone and lowest for fibrous tissue, resp. cystic lesions. The new, more rigorous cleansing process decreased cartilage persistence and overall graft performance. CONCLUSION: Performance of grafts from equine origin was better compared to bovine, ovine and human grafts. The exact reason for this difference was not proven in the current study, but could be related to differences in density of cartilage and subchondral bone between species. PMID- 16356174 TI - Mutations in APC, CTNNB1 and K-ras genes and expression of hMLH1 in sporadic colorectal carcinomas from the Netherlands Cohort Study. AB - BACKGROUND: The early to intermediate stages of the majority of colorectal tumours are thought to be driven by aberrations in the Wnt (APC, CTNNB1) and Ras (K-ras) pathways. A smaller proportion of cancers shows mismatch repair deficiency. The aim of this study was to analyse the co-occurrence of these genetic alterations in relation to tumour and patient characteristics. METHODS: In a group of 656 unselected sporadic colorectal cancer patients, aberrations in the APC, K-ras, CTNNB1 genes, and expression of hMLH1 were investigated. Additionally, tumours were divided in groups based on molecular features and compared with respect to patient's age at diagnosis, sex, family history of colorectal cancer, tumour sub-localisation, Dukes' stage and differentiation. RESULTS: Mutations at the phosphorylation sites (codons 31, 33, 37, and 45) in the CTNNB1 gene were observed in tumours from only 5/464 patients. Tumours with truncating APC mutations and activating K-ras mutations in codons 12 and 13 occurred at similar frequencies (37% (245/656) and 36% (235/656), respectively). Seventeen percent of tumours harboured both an APC and a K-ras mutation (109/656). Nine percent of all tumours (58/656) lacked hMLH1 expression. Patients harbouring a tumour with absent hMLH1 expression were older, more often women, more often had proximal colon tumours that showed poorer differentiation when compared to patients harbouring tumours with an APC and/or K-ras mutation. CONCLUSION: CTNNB1 mutations seem to be of minor importance in sporadic colorectal cancer. The main differences in tumour and patient characteristics are found between groups of patients based on mismatch repair deficiency. PMID- 16356175 TI - Clinical benefit of metaphase I oocytes. AB - BACKGROUND: We studied the benefit of using in vitro matured metaphase I (MI) oocytes for ICSI in patients with a maximum of 6 mature metaphase II (MII) oocytes at retrieval. METHODS: In 2004, 187 ICSI cycles were selected in which maximum 6 MII oocytes and at least one MI oocyte were retrieved. MI oocytes were put in culture to mature until the moment of ICSI, which was performed between 2 to 11 hours after oocyte retrieval (day 0). In exceptional cases, when the patient did not have any mature oocyte at the scheduled time of ICSI, MI oocytes were left to mature overnight and were injected between 19 to 26 hours after retrieval (day 1). Embryos from MI oocytes were chosen for transfer only when no other good quality embryos from MII oocytes were available. Outcome parameters were time period of in vitro maturation (IVM), IVM and fertilization rates, embryo development, clinical pregnancy rates, implantation rates and total MI oocyte utilization rate. RESULTS: The overall IVM rate was 43%. IVM oocytes had lower fertilization rates compared to in vivo matured sibling oocytes (52% versus 68%, P < 0.05). The proportion of poor quality embryos was significantly higher in IVM derived oocytes. One pregnancy and live birth was obtained out of 13 transfers of embryos exclusively derived from IVM oocytes. This baby originated from an oocyte that was injected after 22 hrs of IVM. CONCLUSION: Fertilization of in vitro matured MI oocytes can result in normal embryos and pregnancy, making IVM worthwhile, particularly when few MII oocytes are obtained at retrieval. PMID- 16356176 TI - The economic value of an improved malaria treatment programme in Zambia: results from a contingent valuation survey. AB - BACKGROUND: Zambia is facing a double crisis of increasing malaria burden and dwindling capacity to deal with the endemic malaria burden. The pursuit of sustainable but equity mechanisms for financing malaria programmes is a subject of crucial policy discussion. This requires that comprehensive accounting of the economic impact of the various malaria programmes. Information on the economic value of programmes is essential in soliciting appropriate funding allocations for malaria control. AIMS AND OBJECTIVES: This paper specifically seeks to elicit a measure of the economic benefits of an improved malaria treatment programme in Zambia. The paper also studies the equity implications in malaria treatment given that demand or malaria treatment is determined by household socio-economic status. METHODS: A contingent valuation survey of about 300 Zambian households was conducted in four districts. Willingness-to-pay (WTP) was elicited for an improved treatment programme for malaria in order to generate a measure of the economic benefits of the programme. The payment card method was used in eliciting WTP bids. FINDINGS: The study reports that malaria treatment has significant economic benefits to society. The total economic benefits of an improved treatment programme were estimated at an equivalent of USD 77 million per annum, representing about 1.8% of Zambia's GDP. The study also reports the theoretically anticipated association between WTP and several socio-economic factors. Our income elasticity of demand is positive and similar in magnitude to estimates reported in similar studies. Finally, from an equity standpoint, the constraints imposed by income and socio-economic status are discussed. PMID- 16356177 TI - Socio-economic inequity in demand for insecticide-treated nets, in-door residual house spraying, larviciding and fogging in Sudan. AB - BACKGROUND: In order to optimally prioritize and use public and private budgets for equitable malaria vector control, there is a need to determine the level and determinants of consumer demand for different vector control tools. OBJECTIVES: To determine the demand from people of different socio-economic groups for indoor residual house-spraying (IRHS), insecticide-treated nets (ITNs), larviciding with chemicals (LWC), and space spraying/fogging (SS) and the disease control implications of the result. METHODS: Ratings and levels of willingness-to-pay (WTP) for the vector control tools were determined using a random cross-sectional sample of 720 householdes drawn from two states. WTP was elicited using the bidding game. An asset-based socio-economic status (SES) index was used to explore whether WTP was related to SES of the respondents. RESULTS: IRHS received the highest proportion of highest preferred rating (41.0%) followed by ITNs (23.1%). However, ITNs had the highest mean WTP followed by IRHS, while LWC had the least. The regression analysis showed that SES was positively and statistically significantly related to WTP across the four vector control tools and that the respondents' rating of IRHS and ITNs significantly explained their levels of WTP for the two tools. CONCLUSION: People were willing to pay for all the vector-control tools, but the demand for the vector control tools was related to the SES of the respondents. Hence, it is vital that there are public policies and financing mechanisms to ensure equitable provision and utilisation of vector control tools, as well as protecting the poor from cost-sharing arrangements. PMID- 16356178 TI - Chronic fatigue syndrome--a clinically empirical approach to its definition and study. AB - BACKGROUND: The lack of standardized criteria for defining chronic fatigue syndrome (CFS) has constrained research. The objective of this study was to apply the 1994 CFS criteria by standardized reproducible criteria. METHODS: This population-based case control study enrolled 227 adults identified from the population of Wichita with: (1) CFS (n = 58); (2) non-fatigued controls matched to CFS on sex, race, age and body mass index (n = 55); (3) persons with medically unexplained fatigue not CFS, which we term ISF (n = 59); (4) CFS accompanied by melancholic depression (n = 27); and (5) ISF plus melancholic depression (n = 28). Participants were admitted to a hospital for two days and underwent medical history and physical examination, the Diagnostic Interview Schedule, and laboratory testing to identify medical and psychiatric conditions exclusionary for CFS. Illness classification at the time of the clinical study utilized two algorithms: (1) the same criteria as in the surveillance study; (2) a standardized clinically empirical algorithm based on quantitative assessment of the major domains of CFS (impairment, fatigue, and accompanying symptoms). RESULTS: One hundred and sixty-four participants had no exclusionary conditions at the time of this study. Clinically empirical classification identified 43 subjects as CFS, 57 as ISF, and 64 as not ill. There was minimal association between the empirical classification and classification by the surveillance criteria. Subjects empirically classified as CFS had significantly worse impairment (evaluated by the SF-36), more severe fatigue (documented by the multidimensional fatigue inventory), more frequent and severe accompanying symptoms than those with ISF, who in turn had significantly worse scores than the not ill; this was not true for classification by the surveillance algorithm. CONCLUSION: The empirical definition includes all aspects of CFS specified in the 1994 case definition and identifies persons with CFS in a precise manner that can be readily reproduced by both investigators and clinicians. PMID- 16356179 TI - Tango's maximized excess events test with different weights. AB - BACKGROUND: Tango's maximized excess events test (MEET) has been shown to have very good statistical power in detecting global disease clustering. A nice feature of this test is that it considers a range of spatial scale parameters, adjusting for the multiple testing. This means that it has good power to detect a wide range of clustering processes. The test depends on the functional form of a weight function, and it is unknown how sensitive the test is to the choice of this weight function and what function provides optimal power for different clustering processes. In this study, we evaluate the performance of the test for a wide range of weight functions. RESULTS: The power varies greatly with different choice of weight. Tango's original choice for the weight function works very well. There are also other weight functions that provide good power. CONCLUSION: We recommend the use of Tango's MEET to test global disease clustering, either with the original weight or one of the alternate weights that have good power. PMID- 16356180 TI - Subacute sclerosing panencephalitis: results of the Canadian Paediatric Surveillance Program and review of the literature. AB - BACKGROUND: Subacute Sclerosing Panencephalitis (SSPE) is so rare in developed countries with measles immunization programs that national active surveillance is now needed to capture sufficient number of cases for meaningful analysis of data. Through the Canadian Paediatric Surveillance Program (CPSP), the SSPE study was able to document a national incidence and determine the epidemiology of affected Canadian children. METHODS: Between 1997 and 2000, the CPSP surveyed monthly 1978 to 2294 Canadian pediatricians and sub-specialists for SSPE cases. The response rate varied from 82-86% over those years. RESULTS: Altogether, four SSPE cases were reported to the CPSP: one case before, two during and one after the study period. The incidence of SSPE in Canadian children was 0.06/million children/year. Of the four cases, diagnosed between ages four and 17 years, three children had measles infection in infancy. All children showed a progressive course of dementia, loss of motor skills and epilepsy. Two children were treated with isoprinosine and intraventricular interferon but died in less than three years from disease onset. One child did not have any treatment and died after seven years of illness. One child received intraventricular ribavirin and remains alive, but markedly impaired, nine years following diagnosis. CONCLUSION: The CPSP has demonstrated that Canadian paediatricians and paediatric neurologists may encounter cases of SSPE. This report highlights the clinical course of affected Canadian children and provides a review of the disease and its management. PMID- 16356181 TI - A long-term survivor of Bland-White-Garland syndrome with systemic collateral supply: a case report and review of the literature. AB - BACKGROUND: Bland-White-Garland syndrome (anomalous origin of the left coronary artery from the pulmonary artery) is a rare disease which may result in myocardial infarction, congestive heart failure and sometimes death during the early infantile period. CASE PRESENTATION: A successfully treated case of a 45 year-old mother of 2 children with Bland-White-Garland syndrome and concomitant severe mitral regurgitation is presented. Subsequent therapy consisted of ligation of the anomalous origin of the left coronary artery, anastomosis of the left internal mammary artery to the left anterior descending branch and mitral valve replacement. Continuous blood flow from the left coronary artery ostium during extracorporeal circulation and aorta clamping suggested systemic collateral supply. CONCLUSIONS: Recognition and diagnosis of Bland-White-Garland syndrome is important due to its potentially life-threatening complications. PMID- 16356182 TI - Promoting physical activity using an activity monitor and a tailored web-based advice: design of a randomized controlled trial [ISRCTN93896459]. AB - BACKGROUND: Ageing is associated with a decrease in physical activity. This decrease particularly occurs during specific transitional life stages. Especially during adolescence and young adulthood a steep decrease in physical activity is observed. Inactive people are often not aware of their inactivity. Providing feedback on the actual physical activity level by an activity monitor can increase awareness and may in combination with an individually tailored physical activity advice stimulate a physically active lifestyle. METHODS: In a randomized controlled trial the effectiveness of providing an activity monitor in combination with a personal physical activity advice through the Internet will be examined. Outcome measures are level of physical activity, determinants of physical activity, quality of life, empowerment, aerobic fitness and body composition. Participants are relatively inactive adolescents and young adults who are measured at baseline, after 3 months intervention and 5 months after the end of the intervention. In addition, facilitating and hindering factors for implementation of the intervention will be investigated. DISCUSSION: The use of a personal activity monitor in combination with web-based assisted individually tailored health promotion offers a good opportunity to work interactively with large groups of adolescents and young adults and provide them with advice based on their actual activity level. It has great potential to motivate people to change their behaviour and to our knowledge has not been evaluated before. PMID- 16356183 TI - Use of a food frequency questionnaire in American Indian and Caucasian pregnant women: a validation study. AB - BACKGROUND: Food frequency questionnaires (FFQs) have been validated in pregnant women, but few studies have focused specifically on low-income women and minorities. The purpose of this study was to examine the validity of the Harvard Service FFQ (HSFFQ) among low-income American Indian and Caucasian pregnant women. METHODS: The 100-item HSFFQ was administered three times to a sample of pregnant women, and two sets of 24-hour recalls (six total) were collected at approximately 12 and 28 weeks of gestation. The sample included a total of 283 pregnant women who completed Phase 1 of the study and 246 women who completed Phase 2 of the study. Deattenuated Pearson correlation coefficients were used to compare intakes of 24 nutrients estimated from the second and third FFQ to average intakes estimated from the week-12 and week-28 sets of diet recalls. RESULTS: Deattenuated correlations ranged from 0.09 (polyunsaturated fat) to 0.67 (calcium) for Phase 1 and from 0.27 (sucrose) to 0.63 (total fat) for Phase 2. Average deattenuated correlations for the two phases were 0.48 and 0.47, similar to those reported among other groups of pregnant women. CONCLUSION: The HSFFQ is a simple self-administered questionnaire that is useful in classifying low-income American Indian and Caucasian women according to relative dietary intake during pregnancy. Its use as a research tool in this population may provide important information about associations of nutrient intakes with pregnancy outcomes and may help to identify groups of women who would benefit most from nutritional interventions. PMID- 16356184 TI - Sequence analysis of Drd2, Drd4, and Dat1 in SHR and WKY rat strains. AB - BACKGROUND: The Spontaneously Hypertensive Rat (SHR) shows a number of behaviours that closely parallel those seen in children with attention-deficit hyperactivity disorder. These include motor hyperactivity, excessive responses under a fixed interval/extinction schedule, difficulty in acquiring operant tasks and increased sensitivity to immediate behavioural reinforcement. As in children with ADHD, the behavioural and cognitive deficits in the SHR are responsive to stimulants, including d-amphetamine and d,l-methylphenidate. The non-hyperactive Wistar Kyoto (WKY) rat strain is often used as a control in behavioural studies of the SHR, and WKY itself has been suggested to be a useful animal model of depression. Numerous studies have shown that dopaminergic neurotransmission is altered between the two strains. Human genetic studies have found associations between several dopaminergic genes and both ADHD and depression. METHODS: We sequenced three candidate dopaminergic genes (Drd2, Drd4, and Dat1) in the SHR and WKY to identify between-strain sequence differences. RESULTS: No between-strain sequence differences were found in either Drd2 or Drd4, but several variations were found in the Dat1 gene that encodes the dopamine transporter. CONCLUSION: It is plausible that DNA sequence changes in the Dat1 gene account for some of the behavioural differences observed between the SHR and WKY strains. Future work will focus on elucidating the functional effects of the observed polymorphisms. PMID- 16356185 TI - In situ studies of algal biomass in relation to physicochemical characteristics of the Salt Plains National Wildlife Refuge, Oklahoma, USA. AB - This is the first in a series of experiments designed to characterize the Salt Plains National Wildlife Refuge (SPNWR) ecosystem in northwestern Oklahoma and to catalogue its microbial inhabitants. The SPNWR is the remnant of an ancient ocean, encompassing approximately 65 km2 of variably hypersaline flat land, fed by tributaries of the Arkansas River. Relative algal biomass (i.e., chlorophyll concentrations attributed to Chlorophyll-a-containing oxygenic phototrophs) and physical and chemical parameters were monitored at three permanent stations for a one-year period (July 2000 to July 2001) using a nested block design. Salient features of the flats include annual air temperatures that ranged from -10 to 40 degrees C, and similar to other arid/semi-arid environments, 15-20-degree daily swings were common. Shade is absent from the flats system; intense irradiance and high temperatures (air and sediment surface) resulted in low water availability across the SPNWR, with levels of only ca. 15 % at the sediment surface. Moreover, moderate daily winds were constant (ca. 8-12 km h-1), sometimes achieving maximum speeds of up to 137 km h-1. Typical of freshwater systems, orthophosphate (PO(4)3 ) concentrations were low, ranging from 0.04 to <1 microM; dissolved inorganic nitrogen levels were high, but spatially variable, ranging from ca. 250-600 microM (NO(3)- + NO(2)-) and 4-166 microM (NH(4)+). Phototroph abundance was likely tied to nutrient availability, with high-nutrient sites exhibiting high Chl-a levels (ca. 1.46 mg m-2). Despite these harsh conditions, the phototrophic microbial community was unexpectedly diverse. Preliminary attempts to isolate and identify oxygenic phototrophs from SPNWR water and soil samples yielded 47 species from 20 taxa and 3 divisions. Our data indicate that highly variable, extreme environments might support phototrophic microbial communities characterized by higher species diversity than previously assumed. PMID- 16356186 TI - Effects of psycho-educational intervention on health-related quality of life (QOL) of patients with chronic liver disease referring to Shiraz University of Medical Sciences. AB - BACKGROUND: Chronic liver diseases (CLDs) are progressive disorder which has a significant impact on the well-being of patients and leads to significant morbidity. CLDs are characterized by disturbances in physical, psychological and social aspects of well-being. It causes significant health-related quality of life (QOL) impairment. Psycho-educational interventions targeting to functional factors could be beneficial for patients with CLDs. METHODS: An interventional study was conducted on 110 patients with CLDs in Shiraz Liver Transplantation Center (SLTC). Subjects with the required CLDs criteria were selected and randomly divided into experimental (55) and control (55) groups. A two part questionnaire with 25 items concerning demographic and general information and 29 items regarding QOL was used. The psycho-educational needs of the experimental group were assessed in a session before the intervention, then the experimental group took part in 3 sessions individually and one session in groups. The questionnaires were filled in again for both groups but the control group did not receive the intervention program. The questionnaires were filled in again for both groups one day and three months after the intervention. RESULTS: Findings revealed no significant differences between the two groups from the view point of demographic characteristics such as marital status, gender, etc... (p > 0.05) and from the point of clinical variables no statistically significant differences were found (p > 0.05). Findings revealed statistically significant differences in all domain of QOL in the experimental group three months after the intervention (p = 0.001), while there was no statistically significant differences in the control group. CONCLUSION: The results of this study revealed that psycho educational intervention had a significant effect on QOL of patients with CLDs. PMID- 16356187 TI - Genetic mechanisms of knee osteoarthritis: a population-based longitudinal study. AB - To describe the differences in knee structure and non-knee structural factors between offspring having at least one parent with a total knee replacement for severe primary knee osteoarthritis and age- and sex-matched controls with no family history of knee osteoarthritis, a population-based longitudinal study of 163 matched pairs (mean age 45 years, range 26 to 61) was performed at baseline and about 2 years later. Knee cartilage defect score (0 to 4), cartilage volume and bone size were determined with T1-weighted fat saturation magnetic resonance imaging. Body mass index (BMI), lower-limb muscle strength, knee pain, physical work capacity at 170 beats/minute (PWC170) and radiographic osteoarthritis were measured by standard protocols. In comparison with controls, offspring had higher annual knee cartilage loss (-3.1% versus -2.0% at medial tibial site, -1.9% versus -1.1% at lateral tibial site and -4.7% versus -3.7% at patellar site, all P < 0.05), a greater increase in medial cartilage defect score (+0.15 versus 0.01, P < 0.05) and a greater decline in PWC170 (-0.7 watts/kg versus -0.4 watts/kg, P < 0.01). There were no significant differences in change in BMI, lower-limb muscle strength, knee pain or tibial bone area between these two groups; however, the differences in knee cartilage loss and cartilage defect change decreased in magnitude and became non-significant after adjustment for baseline cartilage volume, tibial bone area, BMI and knee pain. This longitudinal study suggests that knee cartilage loss, change in cartilage defects and decrease in physical fitness all have roles in the development of knee osteoarthritis, which is most probably polygenic but may reflect a shared environment. Importantly, the cartilage changes are largely dependent on baseline differences in cartilage volume, tibial bone area, BMI and knee pain, suggesting that these factors might have a role in their initiation. PMID- 16356188 TI - Differential direct effects of cyclo-oxygenase-1/2 inhibition on proteoglycan turnover of human osteoarthritic cartilage: an in vitro study. AB - Treatment of osteoarthritis (OA) with nonsteroidal anti-inflammatory drugs (NSAIDs) diminishes inflammation along with mediators of cartilage destruction. However, NSAIDs may exert adverse direct effects on cartilage, particularly if treatment is prolonged. We therefore compared the direct effects of indomethacin, naproxen, aceclofenac and celecoxib on matrix turnover in human OA cartilage tissue. Human clinically defined OA cartilage from five different donors was exposed for 7 days in culture to indomethacin, naproxen, aceclofenac and celecoxib--agents chosen based on their cyclo-oxygenase (COX)-2 selectivity. As a control, SC-560 (a selective COX-1 inhibitor) was used. Changes in cartilage proteoglycan turnover and prostaglandin E2 production were determined. OA cartilage exhibited characteristic proteoglycan turnover. Indomethacin further inhibited proteoglycan synthesis; no significant effect of indomethacin on proteoglycan release was found, and proteoglycan content tended to decrease. Naproxen treatment was not associated with changes in any parameter. In contrast, aceclofenac and, prominently, celecoxib had beneficial effects on OA cartilage. Both were associated with increased proteoglycan synthesis and normalized release. Importantly, both NSAIDs improved proteoglycan content. Inhibition of prostaglandin E2 production indirectly showed that all NSAIDs inhibited COX, with the more COX-2 specific agents having more pronounced effects. Selective COX-1 inhibition resulted in adverse effects on all parameters, and prostaglandin E2 production was only mildly inhibited. NSAIDs with low COX-2/COX-1 selectivity exhibit adverse direct effects on OA cartilage, whereas high COX-2/COX-1 selective NSAIDs did not show such effects and might even have cartilage reparative properties. PMID- 16356189 TI - Analysis of Fcgamma receptor haplotypes in rheumatoid arthritis: FCGR3A remains a major susceptibility gene at this locus, with an additional contribution from FCGR3B. AB - The Fcgamma receptors play important roles in the initiation and regulation of many immunological and inflammatory processes, and genetic variants (FCGR) have been associated with numerous autoimmune and infectious diseases. The data in rheumatoid arthritis (RA) are conflicting and we previously demonstrated an association between FCGR3A and RA. In view of the close molecular proximity with FCGR2A, FCGR2B and FCGR3B, additional polymorphisms within these genes and FCGR haplotypes were examined to refine the extent of association with RA. Biallelic polymorphisms in FCGR2A, FCGR2B and FCGR3B were examined for association with RA in two well characterized UK Caucasian and North Indian/Pakistani cohorts, in which FCGR3A genotyping had previously been undertaken. Haplotype frequencies and linkage disequilibrium were estimated across the FCGR locus and a model-free analysis was performed to determine association with RA. This was followed by regression analysis, allowing for phase uncertainty, to identify the particular haplotype(s) that influences disease risk. Our results reveal that FCGR2A, FCGR2B and FCGR3B were not associated with RA. The haplotype with the strongest association with RA susceptibility was the FCGR3A-FCGR3B 158V-NA2 haplotype (odds ratio 3.18, 95% confidence interval 1.13-8.92 [P = 0.03] for homozygotes compared with all genotypes). The association was stronger in the presence of nodules (odds ratio 5.03, 95% confidence interval 1.44-17.56; P = 0.01). This haplotype was also more common in North Indian/Pakistani RA patients than in control individuals, but not significantly so. Logistic regression analyses suggested that FCGR3A remained the most significant gene at this locus. The increased association with an FCGR3A-FCGR3B haplotype suggests that other polymorphic variants within FCGR3A or FCGR3B, or in linkage disequilibrium with this haplotype, may additionally contribute to disease pathogenesis. PMID- 16356190 TI - 52-kDa Ro/SSA epitopes preferentially recognized by antibodies from mothers of children with neonatal lupus and congenital heart block. AB - Neonatal lupus erythematosus is a rare disorder caused by the transplacental passage of maternal autoantibodies. The 52-kDa Ro/SSA antigen (Ro52) ribonucleoprotein represents an antigenic target strongly associated with the autoimmune response in mothers whose children have neonatal lupus and cardiac conduction disturbances, mainly congenital heart block. The objective of this study was to identify putative Ro52/60-kDa Ro/SSA antigen (Ro60) epitopes associated with neonatal lupus and congenital heart block. The reactivity of IgG antibodies present in the sera from mothers with systemic lupus erythematosus and Sjogren's syndrome and in the sera from asymptomatic mothers (a longitudinal study of 192 samples from 66 subjects) was investigated by ELISA using Ro52, Ro60 and 48-kDa La/SSB antigen proteins, as well as 45 synthetic peptides, 13-24 residues long, of Ro52/Ro60 proteins. One to 19 samples collected before, during and after pregnancy were available for each mother. Forty-three disease controls selected randomly and normal sera were tested in parallel. Although no differences were found between Sjogren's syndrome and asymptomatic mothers of group I, who had at least one infant with neonatal lupus, and of group II, who had healthy babies only, significant differences were observed between lupus mothers from both groups. In the former group of lupus mothers, a significantly higher frequency of antibodies to Ro52 peptides 107-122 and 277-292 was observed. Between 18 and 30 weeks of gestation, the period of risk, there was clearly an elevated level of antibodies reacting with Ro52 peptides 1-13, 277-292 and 365 382. Antibodies to Ro52 peptide 365-382 have been shown previously to cross-react with residues 165-185 of the heart 5-HT4 serotoninergic receptor, and might be pathologically important. The level of these Ro52 antibody subsets decreased at the end of pregnancy and after delivery. IgG antibodies to Ro52 peptides 1-13, 107-122, 277-292 and 365-382 may therefore represent important biomarkers to predict a complication in pregnant lupus women with Ro52 antibodies. PMID- 16356191 TI - Regulatory polymorphisms in extracellular matrix protease genes and susceptibility to rheumatoid arthritis: a case-control study. AB - Many extracellular matrix (ECM) proteases seem to be important in rheumatoid arthritis (RA) and regulation of their transcription levels is a critical mechanism for controlling their activity. We have investigated, therefore, whether the best-characterized single nucleotide polymorphisms (SNPs) affecting transcription of the ECM proteases that have been related with joint pathology are associated with RA susceptibility. Nine SNPs in eight genes were selected by bibliographic search, including SNPs in the genes encoding matrix metalloproteinase (MMP)1, MMP2, MMP3, MMP7, MMP9, MMP13, plasminogen activator, tissue type (PLAT) and PAI-1. They were studied in a case-control setting that included 550 RA patients and 652 controls of Spanish ancestry from a single center. Genotyping was performed by single-base extension. Only two of the nine SNPs showed significant association with RA susceptibility. RA patients showed increased frequencies of the -7351 T allele of the gene encoding PLAT (36.4% versus 32.1% in controls, p = 0.026) and the -1306 T allele of the gene encoding MMP2 (24.5% versus 20.3% in controls, p = 0.013). These two alleles seemed to cooperate according to an additive model with respect to increased RA susceptibility (p = 0.004), and they were the low-expression alleles of the respective SNPs in a PLAT enhancer and the MMP2 promoter. These findings are in agreement with previous data suggesting that these two ECM proteases have a protective role in RA pathology. Confirmation of these associations will be needed to support these hypotheses. The remaining SNPs did not show association, either individually or collectively. Therefore, although regulatory SNPs in ECM proteases did not show any major effect on RA susceptibility, it was possible to find modest associations that, if replicated, will have interesting implications in the understanding of RA pathology. PMID- 16356192 TI - Adalimumab clinical efficacy is associated with rheumatoid factor and anti-cyclic citrullinated peptide antibody titer reduction: a one-year prospective study. AB - Studies on autoantibody production in patients treated with tumor necrosis factor alpha (TNF-alpha) inhibitors reported contradictory results. We investigated in a prospective study the efficacy of a treatment with human monoclonal anti-TNF alpha antibody (adalimumab) in patients with rheumatoid arthritis (RA) and we evaluated the relationship between treatment efficacy and the incidence and titers of disease-associated and non-organ-specific autoantibodies. Fifty-seven patients with RA not responsive to methotrexate and treated with adalimumab were enrolled. Antinuclear, anti-double-stranded(ds)DNA, anti-extractable nuclear antigens, anti-cardiolipin (aCL), anti-beta2 glycoprotein I (anti-beta2GPI) autoantibodies, rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) autoantibodies were investigated at baseline and after 6 and 12 months of follow-up. Comparable parameters were evaluated in a further 55 patients treated with methotrexate only. Treatment with adalimumab induced a significant decrease in RF and anti-CCP serum levels, and the decrease in antibody titers correlated with the clinical response to the therapy. A significant induction of antinuclear autoantibodies (ANA) and IgG/IgM anti-dsDNA autoantibodies were also found in 28% and 14.6% patients, respectively, whereas aCL and anti-beta2GPI autoantibodies were not detected in significant quantities. No association between ANA, anti-dsDNA, aCL and anti-beta2GPI autoantibodies and clinical manifestations was found. Clinical efficacy of adalimumab is associated with the decrease in RF and anti-CCP serum levels that was detected after 24 weeks and remained stable until the 48th week of treatment. Antinuclear and anti-dsDNA autoantibodies, but not anti-phospholipid autoantibodies, can be induced by adalimumab but to a lower extent than in studies with other anti-TNF blocking agents. PMID- 16356193 TI - B lymphocyte stimulator (BLyS) isoforms in systemic lupus erythematosus: disease activity correlates better with blood leukocyte BLyS mRNA levels than with plasma BLyS protein levels. AB - Considerable evidence points to a role for B lymphocyte stimulator (BLyS) overproduction in murine and human systemic lupus erythematosus (SLE). Nevertheless, the correlation between circulating levels of BLyS protein and disease activity in human SLE is modest at best. This may be due to an inadequacy of the former to reflect endogenous BLyS overproduction faithfully, in that steady-state protein levels are affected not just by production rates but also by rates of peripheral utilization and excretion. Increased levels of BLyS mRNA may better reflect increased in vivo BLyS production, and therefore they may correlate better with biologic and clinical sequelae of BLyS overexpression than do circulating levels of BLyS protein. Accordingly, we assessed peripheral blood leukocyte levels of BLyS mRNA isoforms (full-length BLyS and DeltaBLyS) and plasma BLyS protein levels in patients with SLE, and correlated these levels with laboratory and clinical features. BLyS protein, full-length BLyS mRNA, and DeltaBLyS mRNA levels were greater in SLE patients (n = 60) than in rheumatoid arthritis patients (n = 60) or normal control individuals (n = 30). Although full length BLyS and DeltaBLyS mRNA levels correlated significantly with BLyS protein levels in the SLE cohort, BLyS mRNA levels were more closely associated with serum immunoglobulin levels and SLE Disease Activity Index scores than were BLyS protein levels. Moreover, changes in SLE Disease Activity Index scores were more closely associated with changes in BLyS mRNA levels than with changes in BLyS protein levels among the 37 SLE patients from whom repeat blood samples were obtained. Thus, full-length BLyS and DeltaBLyS mRNA levels are elevated in SLE and are more closely associated with disease activity than are BLyS protein levels. BLyS mRNA levels may be a helpful biomarker in the clinical monitoring of SLE patients. PMID- 16356194 TI - THR0921, a novel peroxisome proliferator-activated receptor gamma agonist, reduces the severity of collagen-induced arthritis. AB - THR0921 is a novel peroxisome proliferator-activated receptor gamma (PPARgamma) agonist with potent anti-diabetic properties. Because of the proposed role of PPARgamma in inflammation, we investigated the potential of orally active THR0921 to inhibit the pathogenesis of collagen-induced arthritis (CIA). CIA was induced in DBA/1J mice by the injection of bovine type II collagen in complete Freund's adjuvant on days 0 and 21. Mice were treated with THR0921 (50 mg/kg/day) starting on the day of the booster injection and throughout the remaining study period. Both clinical disease activity scores as well as histological scores of joint destruction were significantly reduced in mice treated with THR0921 compared to untreated mice. Proliferation of isolated spleen cells, as well as circulating levels of IgG antibody to type II collagen, was decreased by THR0921. Moreover, spleen cell production of IFN-gamma, tumor necrosis factor (TNF)-alpha and IL 1beta in response to exposure to lipopolysaccharide or type II collagen was reduced by in vivo treatment with THR0921. Steady state mRNA levels of TNF-alpha, IL-1beta, monocyte chemotactic protein-1 and receptor activator of nuclear factor kappaB ligand (RANKL) in isolated joints were all decreased in mice treated with THR0921. Finally, THR0921 inhibited osteoclast differentiation of bone marrow derived cells stimulated with macrophage colony-stimulating factor and RANKL. In conclusion, THR0921 attenuates collagen-induced arthritis in part by reducing the immune response. As such, PPARgamma may be an important therapeutic target for rheumatoid arthritis. PMID- 16356196 TI - Aging, osteoarthritis and transforming growth factor-beta signaling in cartilage. AB - Osteoarthritis is a common malady of the musculoskeletal system affecting the articular cartilage. The increased frequency of osteoarthritis with aging indicates the complex etiology of this disease, which includes pathophysiology and joint stability including biomechanics. The balance between anabolic morphogens and growth factors and catabolic cytokines is at the crux of the problem of osteoarthritis. One such signal is transforming growth factor-beta (TGF-beta). The impaired TGF-beta signaling has been identified as a culprit in old mice in a recent article in this journal. This commentary places this discovery in the context of anabolic and catabolic signals and articular cartilage homeostasis in the joint. PMID- 16356195 TI - Osteoclasts; culprits in inflammatory osteolysis. AB - Periarticular osteolysis, a crippling complication of rheumatoid arthritis, is the product of enhanced osteoclast recruitment and activation. The osteoclast, which is a member of the monocyte/macrophage family, is the exclusive bone resorptive cell, and its differentiation and activation are under the aegis of a variety of cytokines. Receptor activator of NF-kappaB ligand (RANKL) and macrophage colony-stimulating factor are the essential osteoclastogenic cytokines and are increased in inflammatory joint disease. Tumor necrosis factor-alpha, which perpetrates arthritic bone loss, exerts its osteoclastogenic effect in the context of RANKL with which it synergizes. Achieving an understanding of the mechanisms by which the three cytokines affect the osteoclast has resulted in a number of active and candidate therapeutic targets. PMID- 16356197 TI - Magnetic resonance imaging changes of sacroiliac joints in patients with recent onset inflammatory back pain: inter-reader reliability and prevalence of abnormalities. AB - To study the inter-reader reliability of detecting abnormalities of sacroiliac (SI) joints in patients with recent-onset inflammatory back pain by magnetic resonance imaging (MRI), and to study the prevalence of inflammation and structural changes at various sites of the SI joints. Sixty-eight patients with inflammatory back pain (at least four of the five following criteria: symptom onset before age 40, insidious onset, morning stiffness, duration >3 months, improvement with exercise--or three out of five of these plus night pain) were included (38% male; mean age, 34.9 years [standard deviation 10.3]; 46% HLA-B27 positive; mean symptom duration, 18 months), with symptom duration <2 years. A MRI scan of the SI joints was made in the coronal plane with the following sequences: T1-weighted spin echo, short-tau inversion recovery, T2-weighted fast spin echo with fat saturation, and T1-spin echo with fat saturation after the administration of gadolinium. Both SI joints were scored for inflammation (separately for subchondral bone and bone marrow, joint space, joint capsule, ligaments) as well as for structural changes (erosions, sclerosis, ankylosis), by two observers independently. Agreement between the two readers was analysed by concordance and discordance rates and by kappa statistics. Inflammation was present in 32 SI joints of 22 patients, most frequently located in bone marrow and/or subchondral bone (29 joints in 21 patients). Readers agreed on the presence of inflammation in 85% of the cases in the right SI joint and in 78% of the cases in the left SI joint. Structural changes on MRI were present in 11 patients. Ten of these 11 patients also showed signs of inflammation. Agreement on the presence or absence of inflammation and structural changes of SI joints by MRI was acceptable, and was sufficiently high to be useful in ascertaining inflammatory and structural changes due to sacroiliitis. About one-third of patients with recent-onset inflammatory back pain show inflammation, and about one-sixth show structural changes in at least one SI joint. PMID- 16356198 TI - Polarized subsets of human T-helper cells induce distinct patterns of chemokine production by normal and systemic sclerosis dermal fibroblasts. AB - The role of fibroblasts in inflammatory processes and their cross-talk with T cells is increasingly being recognized. Our aim was to explore the capacity of dermal fibroblasts to produce inflammatory chemokines potentially involved in fibrosis occurring in response to contact with polarized human T cells. Our findings indicate that the program of chemokine production by fibroblasts is differentially regulated depending on the T-helper (Th) cell subset used to activate them. Thus, Th1 and Th2 cells preferentially induced production of IFN gamma inducible protein (IP)-10 and IL-8, respectively, whereas monocyte chemoattractant protein (MCP)-1 was equally induced by both subsets at mRNA and protein levels. Neutralization experiments indicated that membrane-associated tumour necrosis factor-alpha and IL-1 played a major role in the induction of IL 8 and MCP-1 by Th1 and Th2 cells, whereas membrane-associated IFN-gamma (present only in Th1 cells) was responsible, at least in part, for the lower IL-8 and higher IP-10 production induced by Th1 cells. The contributions of tumour necrosis factor-alpha, IL-1 and IFN-alpha were confirmed when fibroblasts were cultured separated in a semipermeable membrane from living T cells activated by CD3 cross-linking. We observed further differences when we explored signal transduction pathway usage in fibroblasts. Pharmacological inhibition of c-Jun N terminal kinase and nuclear factor-kappaB resulted in inhibition of IL-8 mRNA transcription induced by Th1 cells but not that by Th2 cells, whereas inhibition of MEK/ERK (mitogen-activated protein kinase of extracellular signal-regulated kinase/extracellular signal-regulated kinase) and nuclear factor-kappaB resulted in inhibition of MCP-1 mRNA induced by Th2 but not by Th1 cells. Finally, no distinct differences in chemokine production were observed when the responses to T cell contact or to prototypic Th1 and Th2 cytokines were examined in systemic sclerosis versus normal fibroblasts. These findings indicate that fibroblasts have the potential to participate in shaping the inflammatory response through the activation of flexible programs of chemokine production that depend on the Th subset eliciting their response. PMID- 16356199 TI - Control of hyperuricemia in subjects with refractory gout, and induction of antibody against poly(ethylene glycol) (PEG), in a phase I trial of subcutaneous PEGylated urate oxidase. AB - PEG-modified recombinant mammalian urate oxidase (PEG-uricase) is being developed as a treatment for patients with chronic gout who are intolerant of, or refractory to, available therapy for controlling hyperuricemia. In an open-label phase I trial, single subcutaneous injections of PEG-uricase (4 to 24 mg) were administered to 13 such subjects (11 had tophaceous gout), whose plasma uric acid concentration (pUAc) was 11.3 +/- 2.1 mg/dl (mean +/- SD). By day seven after injection of PEG-uricase, pUAc had declined by an average of 7.9 mg/dl and had normalized in 11 subjects, whose mean pUAc decreased to 2.8 +/- 2.2 mg/dl. At doses of 8, 12, and 24 mg, the mean pUAc at 21 days after injection remained no more than 6 mg/dl. In eight subjects, plasma uricase activity was still measurable at 21 days after injection (half-life 10.5 to 19.9 days). In the other five subjects, plasma uricase activity could not be detected beyond ten days after injection; this was associated with the appearance of relatively low-titer IgM and IgG antibodies against PEG-uricase. Unexpectedly, these antibodies were directed against PEG itself rather than the uricase protein. Three PEG antibody positive subjects had injection-site reactions at 8 to 9 days after injection. Gout flares in six subjects were the only other significant adverse reactions, and PEG-uricase was otherwise well tolerated. A prolonged circulating life and the ability to normalize plasma uric acid in markedly hyperuricemic subjects suggest that PEG-uricase could be effective in depleting expanded tissue stores of uric acid in subjects with chronic or tophaceous gout. The development of anti PEG antibodies, which may limit efficacy in some patients, is contrary to the general assumption that PEG is non-immunogenic. PEG immunogenicity deserves further investigation, because it has potential implications for other PEGylated therapeutic agents in clinical use. PMID- 16356200 TI - Heterogeneity of psychophysiological stress responses in fibromyalgia syndrome patients. AB - Dysregulated psychophysiological responses have been observed in patients with fibromyalgia syndrome (FMS), although the results are inconsistent. Surface electromyographic (EMG), systolic and diastolic blood pressure, heart rate (HR), and skin conductance levels (SCLs) were continuously recorded at baseline, and during a series of stress and relaxation tasks in 90 FMS patients and 30 age and sex matched healthy controls (HCs). The patient sample demonstrated lower baseline EMG levels compared to the HCs on all tasks. In contrast, the patients displayed elevated HR and SCL (sympathetic vasomotor and sudomotor indices, respectively) during both stress tasks. A cluster analysis identified four psychophysiological response patterns: 63.3% of HCs showed increased muscle tension and stable cardiovascular responses; 34.8% of FMS patients showed a pattern of increased sympathetic vasomotor reactivity with stable sudomotor and reduced muscular response; 12.2% of FMS patients showed a pattern of increased sympathetic sudomotor reactivity connected with increased sympathetic vasomotor response and reduced muscular response; and, in contrast, 46.7% of FMS patients showed a pattern of parasympathetic vasomotor reactivity and reduced sudomotor as well as muscular response. The identification of low baseline muscle tension in FMS is discrepant with other chronic pain syndromes and suggests that unique psychophysiological features may be associated with FMS. The different psychophysiological response patterns within the patient sample support the heterogeneity of FMS. PMID- 16356201 TI - Partial protection against collagen antibody-induced arthritis in PARP-1 deficient mice. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear DNA-binding protein that participates in the regulation of DNA repair and maintenance of genomic integrity. In addition, PARP-1 has a role in several models of inflammation disease, where its absence or inactivation confers protection. The aim of this study was to analyze the impact of selective PARP-1 suppression in collagen antibody-induced arthritis. We show that PARP-1 deficiency partially reduces the severity of arthritis, although the incidence of disease was similar in control and deficient mice. Decreased clinical scores were accompanied by partial reduction of histopathological findings. Interestingly, quantitative real-time PCR and ELISA analysis revealed that the absence of PARP-1 down-regulated IL 1beta and monocyte chemotactic protein 1 expression in arthritic joints whereas tumor necrosis factor-alpha transcription was not impaired. Our results provide evidence of the contribution of PARP-1 to the progression of arthritis and identify this protein as a potential therapeutic target for the treatment of rheumatoid arthritis. PMID- 16356202 TI - Bench-to-bedside review: early tracheostomy in critically ill trauma patients. AB - A significant proportion of trauma patients require tracheostomy during intensive care unit stay. The timing of this procedure remains a subject of debate. The decision for tracheostomy should take into consideration the risks and benefits of prolonged endotracheal intubation versus tracheostomy. Timing of tracheostomy is also influenced by the indications for the procedure, which include relief of upper airway obstruction, airway access in patients with cervical spine injury, management of retained airway secretions, maintenance of patent airway and airway access for prolonged mechanical ventilation. This review summarizes the potential advantages of tracheostomy versus endotracheal intubation, the different indications for tracheostomy in trauma patients and studies examining early versus late tracheostomy. It also reviews the predictors of prolonged mechanical ventilation, which may guide the decision regarding the timing of tracheostomy. PMID- 16356203 TI - Clinical review: percutaneous dilatational tracheostomy. AB - As the number of critically ill patients requiring tracheotomy for prolonged ventilation has increased, the demand for a procedural alternative to the surgical tracheostomy (ST) has also emerged. Since its introduction, percutaneous dilatational tracheostomies (PDT) have gained increasing popularity. The most commonly cited advantages are the ease of the familiar technique and the ability to perform the procedure at the bedside. It is now considered a viable alternative to (ST) in the intensive care unit. Evaluation of PDT procedural modifications will require evaluation in randomized clinical trials. Regardless of the PDT technique, meticulous preoperative and postoperative management are necessary to maintain the excellent safety record of PDT. PMID- 16356204 TI - Clinical review: a systematic review of corticosteroid use in infections. AB - Traditional teaching suggests that corticosteroids should be avoided during acute infectious episodes for fear of compromising the immune response. However, the outcome benefit shown through steroid administration in early septic shock implies this paranoia may be misplaced. We therefore performed a systematic review of the literature to identify the current strength of evidence for the use of corticosteroids in specified infections, and to make appropriate graded recommendations. PMID- 16356205 TI - Correlation of procalcitonin and C-reactive protein to inflammation, complications, and outcome during the intensive care unit course of multiple trauma patients. AB - BACKGROUND: A comparison of the amount of and the kinetics of induction of procalcitonin (PCT) with that of C-reactive protein (CRP) during various types of and severities of multiple trauma, and their relation to trauma-related complications, was performed. METHODS: Ninety adult trauma patients admitted to the intensive care unit of our tertiary care hospital were evaluated in a prospective case study. During the initial 24 hours after trauma the Injury Severity Score, the Sepsis-related Organ Failure Assessment score, and the Acute Physiology and Chronic Health Evaluation II score were evaluated. PCT, CRP, the sepsis criteria (American College of Chest Physicians/Society of Critical Care Medicine definitions), and the Sepsis-related Organ Failure Assessment score were measured at days 1-7, as well as at days 14 and 21, concluding the observation period with the 28-day survival. RESULTS: The induction of PCT and CRP varied in patients suffering from trauma. PCT increased only moderately in most patients and peaked at day 1-2 after trauma, the concentrations rapidly declining thereafter. CRP ubiquitously increased and its kinetics were much slower. Complications such as sepsis, infection, blood transfusion, prolonged intensive care unit treatment, and poor outcome were more frequent in patients with initially high PCT (>1 ng/ml), whereas increases of CRP showed no positive correlation. CONCLUSION: In patients with multiple trauma due to an accident, the PCT level provides more information than the CRP level since only moderate amounts of PCT are induced, and higher concentrations correlate with more severe trauma and a higher frequency of various complications, including sepsis and infection. Most importantly, the moderate trauma-related increase of PCT and the rapidly declining concentrations provide a baseline value near to the normal range at an earlier time frame than for CRP, thus allowing a faster and more valid prediction of sepsis during the early period after trauma. PMID- 16356206 TI - Bronchoalveolar lavage cytological alveolar damage in patients with severe pneumonia. AB - INTRODUCTION: Histological examination of lung specimens from patients with pneumonia shows the presence of desquamated pneumocytes and erythrophages. We hypothesized that these modifications should also be present in bronchoalveolar lavage fluid (BAL) from patients with hospital-acquired pneumonia. METHODS: We conducted a prospective study in mechanically ventilated patients with clinical suspicion of pneumonia. Patients were classified as having hospital-acquired pneumonia or not, in accordance with the quantitative microbiological cultures of respiratory tract specimens. A group of severe community-acquired pneumonias requiring mechanical ventilation during the same period was used for comparison. A specimen of BAL (20 ml) was taken for cytological analysis. A semiquantitative analysis of the dominant leukocyte population, the presence of erythrophages/siderophages and desquamated type II pneumocytes was performed. RESULTS: In patients with confirmed hospital-acquired pneumonia, we found that 13 out of 39 patients (33.3%) had erythrophages/siderophages in BAL, 18 (46.2%) had desquamated pneumocytes and 8 (20.5%) fulfilled both criteria. Among the patients with community-acquired pneumonia, 7 out of 15 (46.7%) had erythrophages/siderophages and 6 (40%) had desquamated pneumocytes on BAL cytology. Only four (26.7%) fulfilled both criteria. No patient without hospital acquired pneumonia had erythrophages/siderophages and only 3 out of 18 (16.7%) had desquamated pneumocytes on BAL cytology. CONCLUSION: Cytological analysis of BAL from patients with pneumonia (either community-acquired or hospital-acquired) shows elements of cytological alveolar damage as hemorrhage and desquamated type II pneumocytes much more frequently than in BAL from patients without pneumonia. These elements had a high specificity for an infectious cause of pulmonary infiltrates but low specificity. These lesions could serve as an adjunct to diagnosis in patients suspected of having ventilator-associated pneumonia. PMID- 16356207 TI - A comparison of admission and worst 24-hour Acute Physiology and Chronic Health Evaluation II scores in predicting hospital mortality: a retrospective cohort study. AB - INTRODUCTION: The Acute Physiology and Chronic Health Evaluation (APACHE) II score is widely used in the intensive care unit (ICU) as a scoring system for research and clinical audit purposes. Physiological data for calculation of the APACHE II score are derived from the worst values in the first 24 hours after admission to the ICU. The collection of physiological data on admission only is probably logistically easier, and this approach is used by some ICUs. This study compares the performance of APACHE II scores calculated using admission data with those obtained from the worst values in the first 24 hours. MATERIALS AND METHODS: This was a retrospective cohort study using prospectively collected data from a tertiary ICU. There were no missing physiological data and follow-up for mortality was available for all patients in the database. The admission and the worst 24-hour physiological variables were used to generate the admission APACHE II score and the worst 24-hour APACHE II score, and the corresponding predicted mortality, respectively. RESULTS: There were 11,107 noncardiac surgery ICU admissions during 11 years from 1 January 1993 to 31 December 2003. The mean admission and the worst 24-hour APACHE II score were 12.7 and 15.4, and the derived predicted mortality estimates were 15.5% and 19.3%, respectively. The actual hospital mortality was 16.3%. The overall discrimination ability, as measured by the area under the receiver operating characteristic curve, of the admission APACHE II model (83.8%, 95% confidence interval = 82.9-84.7) and the worst 24-hour APACHE II model (84.6%, 95% confidence interval = 83.7-85.5) was not significantly different (P = 1.00). CONCLUSION: Substitution of the worst 24 hour physiological variables with the admission physiological variables to calculate the admission APACHE II score maintains the overall discrimination ability of the traditional APACHE II model. The admission APACHE II model represents a potential alternative model to the worst 24-hour APACHE II model in critically ill nontrauma patients. PMID- 16356208 TI - A preliminary study on the monitoring of mixed venous oxygen saturation through the left main bronchus. AB - INTRODUCTION: The study sought to assess the feasibility and accuracy of measuring mixed venous oxygen saturation (SvO2) through the left main bronchus (SpO2(trachea)) METHODS: Twenty hybrid pigs of each sex were studied. After anesthesia, a Robertshaw double-lumen tracheal tube with a single-use pediatric pulse oximeter attached to the left lateral surface was introduced toward the left main bronchus of the pig by means of a fibrobronchoscope. Measurements of SpO2(trachea) and oxygen saturation from pulmonary artery samples (SvO2(blood)) were performed with an intracuff pressure of 0 to 60 cmH2O. After equilibration, hemorrhagic shock was induced in these pigs by bleeding to a mean arterial blood pressure of 40 mmHg. With the intracuff pressure maintained at 60 cmH2O, SpO2(trachea) and SvO2(blood) were obtained respectively during the pre-shock period, immediately after the onset of shock, 15 and 30 minutes after shock, and 15, 30, and 60 minutes after resuscitation. RESULTS: SpO2(trachea) was the same as SvO2(blood) at an intracuff pressure of 10, 20, 40, and 60 cmH2O, but was reduced when the intracuff pressure was zero (p < 0.001 compared with SvO2(blood)) in hemodynamically stable states. Changes of SpO2(trachea) and SvO2(blood) corresponded with varieties of cardiac output during the hemorrhagic shock period. There was a significant correlation between the two methods at different time points. CONCLUSION: Measurement of the left main bronchus SpO2 is feasible and provides similar readings to SvO2(blood) in hemodynamically stable or in low saturation states. Tracheal oximetry readings are not primarily derived from the tracheal mucosa. The technique merits further evaluation. PMID- 16356209 TI - Using simulation for training and to change protocol during the outbreak of severe acute respiratory syndrome. AB - INTRODUCTION: During the 2003 severe acute respiratory syndrome (SARS) crisis, we proposed and tested a new protocol for cardiac arrest in a patient with SARS. The protocol was rapidly and effectively instituted by teamwork training using high fidelity simulation. METHODS: Phase 1 was a curriculum design of a SARS-specific cardiac arrest protocol in three steps: planning the new protocol, repeated simulations of this protocol in a classroom, and a subsequent simulation of a cardiac arrest on a hospital ward. Phase 2 was the training of 275 healthcare workers (HCWs) using the new protocol. Training involved a seminar, practice in wearing the mandatory personal protection system (PPS), and cardiac arrest simulations with subsequent debriefing. RESULTS: Simulation provided insights that had not been considered in earlier phases of development. For example, a single person can don a PPS worn for the SARS patient in 1 1/2 minutes. However, when multiple members of a cardiac arrest team were dressing simultaneously, the time to don the PPS increased to between 3 1/2 and 5 1/2 minutes. Errors in infection control as well as in medical management of advanced cardiac life support (ACLS) were corrected. CONCLUSION: During the SARS crisis, real-time use of a high-fidelity simulator allowed the training of 275 HCWs in 2 weeks, with debriefing and error management. HCWs were required to manage the SARS cardiac arrest wearing unfamiliar equipment and following a modified ACLS protocol. The insight gained from this experience will be valuable for future infectious disease challenges in critical care. PMID- 16356210 TI - Bench-to-bedside review: weaning failure--should we rest the respiratory muscles with controlled mechanical ventilation? AB - The use of controlled mechanical ventilation (CMV) in patients who experience weaning failure after a spontaneous breathing trial or after extubation is a strategy based on the premise that respiratory muscle fatigue (requiring rest to recover) is the cause of weaning failure. Recent evidence, however, does not support the existence of low frequency fatigue (the type of fatigue that is long lasting) in patients who fail to wean despite the excessive respiratory muscle load. This is because physicians have adopted criteria for the definition of spontaneous breathing trial failure and thus termination of unassisted breathing, which lead them to put patients back on the ventilator before the development of low frequency respiratory muscle fatigue. Thus, no reason exists to completely unload the respiratory muscles with CMV for low frequency fatigue reversal if weaning is terminated based on widely accepted predefined criteria. This is important, since experimental evidence suggests that CMV can induce dysfunction of the diaphragm, resulting in decreased diaphragmatic force generating capacity, which has been called ventilator-induced diaphragmatic dysfunction (VIDD). The mechanisms of VIDD are not fully elucidated, but include muscle atrophy, oxidative stress and structural injury. Partial modes of ventilatory support should be used whenever possible, since these modes attenuate the deleterious effects of mechanical ventilation on respiratory muscles. When CMV is used, concurrent administration of antioxidants (which decrease oxidative stress and thus attenuate VIDD) seems justified, since antioxidants may be beneficial (and are certainly not harmful) in critical care patients. PMID- 16356211 TI - Evaluating mortality in intensive care units: contribution of competing risks analyses. AB - INTRODUCTION: Kaplan-Meier curves and logistic models are widely used to describe and explain the variability of survival in intensive care unit (ICU) patients. The Kaplan-Meier approach considers that patients discharged alive from hospital are 'non-informatively' censored (for instance, representative of all other individuals who have survived to that time but are still in hospital); this is probably wrong. Logistic models are adapted to this so-called 'competing risks' setting but fail to take into account censoring and differences in exposure time. To address these issues, we exemplified the usefulness of standard competing risks methods; namely, cumulative incidence function (CIF) curves and the Fine and Gray model. METHODS: We studied 203 mechanically ventilated cancer patients with acute respiratory failure consecutively admitted over a five-year period to a teaching hospital medical ICU. Among these patients, 97 died before hospital discharge. After estimating the CIF of hospital death, we used Fine and Gray models and logistic models to explain variability hospital mortality. RESULTS: The CIF of hospital death was 35.5% on day 14 and was 47.8% on day 60 (97/203); there were no further deaths. Univariate models, either the Fine and Gray model or the logistic model, selected the same eight variables as carrying independent information on hospital mortality at the 5% level. Results of multivariate were close, with four variables selected by both models: autologous stem cell transplantation, absence of congestive heart failure, neurological impairment, and acute respiratory distress syndrome. Two additional variables, clinically documented pneumonia and the logistic organ dysfunction, were selected by the Fine and Gray model. CONCLUSION: The Fine and Gray model appears of interest when predicting mortality in ICU patients. It is closely related to the logistic model, through direct modeling of times to death, and can be easily extended to model non-fatal outcomes. PMID- 16356212 TI - Outcomes of interfacility critical care adult patient transport: a systematic review. AB - INTRODUCTION: We aimed to determine the adverse events and important prognostic factors associated with interfacility transport of intubated and mechanically ventilated adult patients. METHODS: We performed a systematic review of MEDLINE, CENTRAL, EMBASE, CINAHL, HEALTHSTAR, and Web of Science (from inception until 10 January 2005) for all clinical studies describing the incidence and predictors of adverse events in intubated and mechanically ventilated adult patients undergoing interfacility transport. The bibliographies of selected articles were also examined. RESULTS: Five studies (245 patients) met the inclusion criteria. All were case-series and two were prospective in design. Due to the paucity of studies and significant heterogeneity in study population, outcome events, and results, we synthesized data in a qualitative manner. Pre-transport severity of illness was reported in only one study. The most common indication for transport was a need for investigations and/or specialist care (three studies, 220 patients). Transport modalities included air (fixed or rotor wing; 66% of patients) and ground (31%) ambulance, and commercial aircraft (3%). Transport teams included a physician in three studies (220 patients). Death during transfer was rare (n = 1). No other adverse events or significant therapeutic interventions during transport were reported. One study reported a 19% (28/145) incidence of respiratory alkalosis on arrival and another study documented a 30% overall intensive care unit mortality, while no adverse events or outcomes were reported after arrival in the three other studies. CONCLUSION: Insufficient data exist to draw firm conclusions regarding the mortality, morbidity, or risk factors associated with the interfacility transport of intubated and mechanically ventilated adult patients. Further study is required to define the risks and benefits of interfacility transfer in this patient population. Such information is important for the planning and allocation of resources related to transporting critically ill adults. PMID- 16356213 TI - Contemporary management of infected necrosis complicating severe acute pancreatitis. AB - Pancreatic necrosis complicating severe acute pancreatitis is a challenging scenario in contemporary critical care practice; it requires multidisciplinary care in a setting where there is a relatively limited evidence base to support decision making. This commentary provides a concise overview of current management of patients with infected necrosis, focusing on detection, the role of pharmacologic intervention, and the timing and nature of surgical interventions. Fine-needle aspiration of necrosis remains the mainstay for establishment of infection. Pharmacological intervention includes antibiotic therapy as an adjunct to surgical debridement/drainage and, more recently, drotrecogin alfa. Specific concerns remain regarding the suitability of drotrecogin alfa in this setting. Early surgical intervention is unhelpful; surgery is indicated when there is strong evidence for infection of necrotic tissue, with the current trend being toward 'less drastic' surgical interventions. PMID- 16356216 TI - Mechanical ventilation in severe asthma. PMID- 16356217 TI - Role of cardiac troponin as a prognosticator in critically ill patients. PMID- 16356218 TI - Cerebral perfusion pressure and risk of brain hypoxia in severe head injury: a prospective observational study. AB - INTRODUCTION: Higher and lower cerebral perfusion pressure (CPP) thresholds have been proposed to improve brain tissue oxygen pressure (PtiO2) and outcome. We study the distribution of hypoxic PtiO2 samples at different CPP thresholds, using prospective multimodality monitoring in patients with severe traumatic brain injury. METHODS: This is a prospective observational study of 22 severely head injured patients admitted to a neurosurgical critical care unit from whom multimodality data was collected during standard management directed at improving intracranial pressure, CPP and PtiO2. Local PtiO2 was continuously measured in uninjured areas and snapshot samples were collected hourly and analyzed in relation to simultaneous CPP. Other variables that influence tissue oxygen availability, mainly arterial oxygen saturation, end tidal carbon dioxide, body temperature and effective hemoglobin, were also monitored to keep them stable in order to avoid non-ischemic hypoxia. RESULTS: Our main results indicate that half of PtiO2 samples were at risk of hypoxia (defined by a PtiO2 equal to or less than 15 mmHg) when CPP was below 60 mmHg, and that this percentage decreased to 25% and 10% when CPP was between 60 and 70 mmHg and above 70 mmHg, respectively (p < 0.01). CONCLUSION: Our study indicates that the risk of brain tissue hypoxia in severely head injured patients could be really high when CPP is below the normally recommended threshold of 60 mmHg, is still elevated when CPP is slightly over it, but decreases at CPP values above it. PMID- 16356220 TI - Changes in central venous saturation after major surgery, and association with outcome. AB - INTRODUCTION: Despite recent interest in measurement of central venous oxygen saturation (ScvO2), there are no published data describing the pattern of ScvO2 changes after major general surgery or any relationship with outcome. METHODS: ScvO2 and other biochemical, physiological and demographic data were prospectively measured for 8 hours after major surgery. Complications and deaths occurring within 28 days of enrollment were included in the data analysis. Independent predictors of complications were identified with the use of logistic regression analysis. Optimum cutoffs for ScvO2 were identified by receiver operator characteristic analysis. RESULTS: Data from 118 patients was analysed; 123 morbidity episodes occurred in 64 these patients. There were 12 deaths (10.2%). The mean +/- SD age was 66.8 +/- 11.4 years. Twenty patients (17%) underwent emergency surgery and 77 patients (66%) were male. The mean +/- SD P POSSUM (Portsmouth Physiologic and Operative Severity Score for the enUmeration of Mortality and morbidity) score was 38.6 +/- 7.7, with a predicted mortality of 16.7 +/- 17.6%. After multivariate analysis, the lowest cardiac index value (odds ratio (OR) 0.58 (95% confidence intervals 0.37 to 0.9); p = 0.018), lowest ScvO2 value (OR 0.94 (0.89 to 0.98); p = 0.007) and P-POSSUM score (OR 1.09 (1.02 to 1.15); p = 0.008) were independently associated with post-operative complications. The optimal ScvO2 cutoff value for morbidity prediction was 64.4%. In the first hour after surgery, significant reductions in ScvO2 were observed, but there were no significant changes in CI or oxygen delivery index during the same period. CONCLUSION: Significant fluctuations in ScvO2 occur in the immediate post-operative period. These fluctuations are not always associated with changes in oxygen delivery, suggesting that oxygen consumption is also an important determinant of ScvO2. Reductions in ScvO2 are independently associated with post operative complications. PMID- 16356219 TI - Early goal-directed therapy after major surgery reduces complications and duration of hospital stay. A randomised, controlled trial [ISRCTN38797445]. AB - INTRODUCTION: Goal-directed therapy (GDT) has been shown to improve outcome when commenced before surgery. This requires pre-operative admission to the intensive care unit (ICU). In cardiac surgery, GDT has proved effective when commenced after surgery. The aim of this study was to evaluate the effect of post-operative GDT on the incidence of complications and duration of hospital stay in patients undergoing general surgery. METHODS: This was a randomised controlled trial with concealed allocation. High-risk general surgical patients were allocated to post operative GDT to attain an oxygen delivery index of 600 ml min(-1) m(-2) or to conventional management. Cardiac output was measured by lithium indicator dilution and pulse power analysis. Patients were followed up for 60 days. RESULTS: Sixty-two patients were randomised to GDT and 60 patients to control treatment. The GDT group received more intravenous colloid (1,907 SD +/- 878 ml versus 1,204 SD +/- 898 ml; p < 0.0001) and dopexamine (55 patients (89%) versus 1 patient (2%); p < 0.0001). Fewer GDT patients developed complications (27 patients (44%) versus 41 patients (68%); p = 0.003, relative risk 0.63; 95% confidence intervals 0.46 to 0.87). The number of complications per patient was also reduced (0.7 SD +/- 0.9 per patient versus 1.5 SD +/- 1.5 per patient; p = 0.002). The median duration of hospital stay in the GDT group was significantly reduced (11 days (IQR 7 to 15) versus 14 days (IQR 11 to 27); p = 0.001). There was no significant difference in mortality (seven patients (11.3%) versus nine patients (15%); p = 0.59). CONCLUSION: Post-operative GDT is associated with reductions in post-operative complications and duration of hospital stay. The beneficial effects of GDT may be achieved while avoiding the difficulties of pre operative ICU admission. PMID- 16356221 TI - Pulse contour analysis after normothermic cardiopulmonary bypass in cardiac surgery patients. AB - INTRODUCTION: Monitoring of the cardiac output by continuous arterial pulse contour (COPiCCOpulse) analysis is a clinically validated procedure proved to be an alternative to the pulmonary artery catheter thermodilution cardiac output (COPACtherm) in cardiac surgical patients. There is ongoing debate, however, of whether the COPiCCOpulse is accurate after profound hemodynamic changes. The aim of this study was therefore to compare the COPiCCOpulse after cardiopulmonary bypass (CPB) with a simultaneous measurement of the COPACtherm. METHODS: After ethical approval and written informed consent, data of 45 patients were analyzed during this prospective study. During coronary artery bypass graft surgery, the aortic transpulmonary thermodilution cardiac output (COPiCCOtherm) and the COPACtherm were determined in all patients. Prior to surgery, the COPiCCOpulse was calibrated by triple transpulmonary thermodilution measurement of the COPiCCOtherm. After termination of CPB, the COPiCCOpulse was documented. Both COPACtherm and COPiCCOtherm were also simultaneously determined and documented. RESULTS: Regression analysis between COPACtherm and COPiCCOtherm prior to CPB showed a correlation coefficient of 0.95 (P < 0.001), and after CPB showed a correlation coefficient of 0.82 (P < 0.001). Bland-Altman analysis showed a mean bias and limits of agreement of 0.0 l/minute and -1.4 to +1.4 l/minute prior to CPB and of 0.3 l/minute and -1.9 to +2.5 l/minute after CPB, respectively. Regression analysis of COPiCCOpulse versus COPiCCOtherm and of COPiCCOpulse versus COPACtherm after CPB showed a correlation coefficient of 0.67 (P < 0.001) and 0.63 (P < 0.001), respectively. Bland-Altman analysis showed a mean bias and limits of agreement of -1.1 l/minute and -1.9 to +4.1 l/minute versus -1.4 l/minute and -4.8 to +2.0 l/minute, respectively. CONCLUSION: We observed an excellent correlation of COPiCCOtherm and COPACtherm measurement prior to CPB. Pulse contour analysis did not yield reliable results with acceptable accuracy and limits of agreement under difficult conditions after weaning from CPB in cardiac surgical patients. The pulse contour analysis thus should be re calibrated as soon as possible, to prevent false therapeutic consequences. PMID- 16356222 TI - Anti-L-selectin antibody therapy does not worsen the postseptic course in a baboon model. AB - INTRODUCTION: Anti-adhesion molecule therapy prevents leukocytes from extravasating. During exaggerated inflammation, this effect is wanted; however, during infection, blocking diapedesis may be detrimental. In this study, therefore, the potential risks of anti-L-selectin antibody therapy were evaluated in a primate model of sepsis. METHODS: Sixteen baboons were anesthetized and randomized into two groups. The experimental group received 2 mg/kg of the anti-L selectin antibody HuDREG-55 and the control group received Ringer's solution prior to the onset of a 2 h infusion of Escherichia coli (1-2 x 10(9) colony forming units (CFU)/kg body weight). Serial blood samples were drawn over a 72 h period for the measurement of tumour necrosis factor-alpha, IL-6 and polymorphonuclear elastase. In addition, blood gas analysis, hematology and routine clinical chemistry were determined to monitor cardiovascular status, tissue perfusion and organ function. RESULTS: The three-day mortality rate and the mean survival time after E. coli-induced sepsis were similar in the two groups. The bacterial blood CFU levels were significantly higher in the placebo group than in the anti-L-selectin group. Other parameters measured throughout the 72 h experimental period, including the cardiovascular, immunologic, and hematologic responses as well as indicators of organ function and tissue perfusion, were similar in the two groups, with the exception of serum creatinine and mean arterial pressure at 32 h after E. coli challenge. CONCLUSION: Anti-L selectin therapy did not adversely affect survival, promote organ dysfunction or result in major side effects in the baboon sepsis model. Additionally, as anti-L selectin therapy improved the bacterial clearance rate, it appears that this therapy is not detrimental during sepsis. This is in contrast to previous studies using the baboon model, in which antibody therapy used to block CD18 increased mortality. PMID- 16356223 TI - Is albumin administration in the acutely ill associated with increased mortality? Results of the SOAP study. AB - INTRODUCTION: Albumin administration in the critically ill has been the subject of some controversy. We investigated the use of albumin solutions in European intensive care units (ICUs) and its relationship to outcome. METHODS: In a cohort, multicenter, observational study, all patients admitted to one of the participating ICUs between 1 May and 15 May 2002 were followed up until death, hospital discharge, or for 60 days. Patients were classified according to whether or not they received albumin at any time during their ICU stay. RESULTS: Of 3,147 admitted patients, 354 (11.2%) received albumin and 2,793 (88.8%) did not. Patients who received albumin were more likely to have cancer or liver cirrhosis, to be surgical admissions, and to have sepsis. They had a longer length of ICU stay and a higher mortality rate, but were also more severely ill, as manifested by higher simplified acute physiology score (SAPS) II and sequential organ failure assessment (SOFA) scores than the other patients. A Cox proportional hazard model indicated that albumin administration was significantly associated with decreased 30-day survival. Moreover, in 339 pairs matched according to a propensity score, ICU and hospital mortality rates were higher in the patients who had received albumin than in those who had not (34.8 versus 20.9% and 41.3 versus 27.7%, respectively, both p < 0.001). CONCLUSION: Albumin administration was associated with decreased survival in this population of acutely ill patients. Further prospective randomized controlled trials are needed to examine the effects of albumin administration in sub-groups of acutely ill patients. PMID- 16356224 TI - Early veno-venous haemodiafiltration for sepsis-related multiple organ failure. AB - INTRODUCTION: We conducted a prospective observational study from January 1995 to December 2004 to evaluate the impact on recovery of a major advance in renal replacement therapy, namely continuous veno-venous haemodiafiltration (CVVHDF), in patients with refractory septic shock. METHOD: CVVHDF was implemented after 6 12 hours of maximal haemodynamic support, and base excess monitoring was used to evaluate the improvement achieved. Of the 60 patients studied, 40 had improved metabolic acidosis after 12 hours of CVVHDF, with a progressive improvement in all failing organs; the final mortality rate in this subgroup was 30%. In contrast, metabolic acidosis did not improve in the remaining 20 patients after 12 hours of CVVHDF, and the mortality rate in this subgroup was 100%. The crude mortality rate for the whole group was 53%, which is significantly lower than the predicted mortality using Simplified Acute Physiology Score II (79%). CONCLUSION: Early CVVHDF may improve the prognosis of sepsis-related multiple organ failure. Failure to correct metabolic acidosis rapidly during the procedure was a strong predictor of mortality. PMID- 16356225 TI - The impact of compliance with 6-hour and 24-hour sepsis bundles on hospital mortality in patients with severe sepsis: a prospective observational study. AB - INTRODUCTION: Compliance with the ventilator care bundle affects the rate of ventilator-associated pneumonia. It was not known, however, whether compliance with sepsis care bundles has an impact on outcome. The aims of the present study were to determine the rate of compliance with 6-hour and 24-hour sepsis bundles and to determine the impact of the compliance on hospital mortality in patients with severe sepsis or septic shock. METHODS: We conducted a prospective observational study on 101 consecutive adult patients with severe sepsis or septic shock on medical or surgical wards, or in accident and emergency areas at two acute National Health Service Trust Teaching hospitals in England. The main outcome measures were: the rate of compliance with 6-hour and 24-hour sepsis care bundles adapted from the Surviving Sepsis Campaign guidelines on patients' clinical care; and the difference in hospital mortality between the compliant and the non-compliant groups. RESULTS: The median age of the patients was 69 years (interquartile range 51 to 78), and 53% were male. The sources of infection were sought and confirmed in 87 of 101 patients. The chest was the most common source (50%), followed by the abdomen (22%). The rate of compliance with the 6-hour sepsis bundle was 52%. Compared with the compliant group, the non-compliant group had a more than twofold increase in hospital mortality (49% versus 23%, relative risk (RR) 2.12 (95% confidence interval (CI) 1.20 to 3.76), P = 0.01) despite similar age and severity of sepsis. Compliance with the 24-hour sepsis bundle was achieved in only 30% of eligible candidates (21/69). Hospital mortality was increased in the non-compliant group from 29% to 50%, with a 76% increase in risk for death, although the difference did not reach statistical significance (RR 1.76 (95% CI 0.84 to 3.64), P = 0.16). CONCLUSION: Non-compliance with the 6-hour sepsis bundle was associated with a more than twofold increase in hospital mortality. Non-compliance with the 24-hour sepsis bundle resulted in a 76% increase in risk for hospital death. All medical staff should practise these relatively simple, easy and cheap bundles within a strict timeframe to improve survival rates in patients with severe sepsis and septic shock. PMID- 16356226 TI - Meta-analysis of hemodynamic optimization: relationship to methodological quality. AB - INTRODUCTION: To review systematically the effect of interventions aimed at hemodynamic optimization and to relate this to the quality of individual published trials. METHODS: A systematic, computerized bibliographic search of published studies and citation reviews of relevant studies was performed. All randomized clinical trials in which adult patients were included in a trial deliberately aiming at an optimized or maximized hemodynamic condition of the patients (with oxygen delivery, cardiac index, oxygen consumption, mixed venous oxygen saturation and/or stroke volume as end-points) were selected. A total of 30 studies were selected for independent review. Two reviewers extracted data on population, intervention, outcome and methodological quality. Agreement between reviewers was high: differences were eventually resolved by third-party decision. The methodological quality of the studies was moderate (mean 9.0, SD 1.7), and the outcomes of the randomized clinical trials were not related to their quality. RESULTS: Efforts to achieve an optimized hemodynamic condition resulted in a decreased mortality rate (relative risk ratio (RR) 0.75 (95% confidence interval (CI) 0.62 to 0.90) in all studies combined. This was due to a significantly decreased mortality in peri-operative intervention studies (RR 0.66 (95% CI 0.54 to 0.81). Overall, patients with sepsis and overt organ failure do not benefit from this method (RR 0.92 (95% CI 0.75 to 1.11)). CONCLUSION: This systematic review showed that interventions aimed at hemodynamic optimization reduced mortality. In particular, trials including peri-operative interventions aimed at the hemodynamic optimization of high-risk surgical patients reduce mortality. Overall, this effect was not related to the trial quality. PMID- 16356227 TI - Spontaneous breathing with airway pressure release ventilation favors ventilation in dependent lung regions and counters cyclic alveolar collapse in oleic-acid induced lung injury: a randomized controlled computed tomography trial. AB - INTRODUCTION: Experimental and clinical studies have shown a reduction in intrapulmonary shunt with spontaneous breathing during airway pressure release ventilation (APRV) in acute lung injury. This reduction was related to reduced atelectasis and increased aeration. We hypothesized that spontaneous breathing will result in better ventilation and aeration of dependent lung areas and in less cyclic collapse during the tidal breath. METHODS: In this randomized controlled experimental trial, 22 pigs with oleic-acid-induced lung injury were randomly assigned to receive APRV with or without spontaneous breathing at comparable airway pressures. Four hours after randomization, dynamic computed tomography scans of the lung were obtained in an apical slice and in a juxtadiaphragmatic transverse slice. Analyses of regional attenuation were performed separately in nondependent and dependent halves of the lungs on end expiratory scans and end-inspiratory scans. Tidal changes were assessed as differences between inspiration and expiration of the mechanical breaths. RESULTS: Whereas no differences were observed in the apical slices, spontaneous breathing resulted in improved tidal ventilation of dependent lung regions (P < 0.05) and less cyclic collapse (P < 0.05) in the juxtadiaphragmatic slices. In addition, with spontaneous breathing, the end-expiratory aeration increased and nonaerated tissue decreased in dependent lung regions close to the diaphragm (P < 0.05 for the interaction ventilator mode and lung region). CONCLUSION: Spontaneous breathing during APRV redistributes ventilation and aeration to dependent, usually well-perfused, lung regions close to the diaphragm, and may thereby contribute to improved arterial oxygenation. Spontaneous breathing also counters cyclic collapse, which is a risk factor for ventilation-associated lung injury. PMID- 16356228 TI - Intensive insulin therapy does not alter the inflammatory response in patients undergoing coronary artery bypass grafting: a randomized controlled trial [ISRCTN95608630]. AB - INTRODUCTION: Strict control of plasma glucose in diabetic and non-diabetic patients has been shown to improve outcome in several clinical settings. There is extensive evidence that glucose can stimulate the production of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-alpha and IL-6, with no effect on the anti-inflammatory cytokine IL-10. We hypothesized that strict glucose regulation results in a change in cytokine balance from a pro-inflammatory state to a more balanced anti-inflammatory condition. In a randomized controlled trial we studied the effect of strict glycemic control on the local and systemic pro inflammatory and anti-inflammatory balance in non-diabetic patients undergoing elective coronary artery bypass grafting with cardiopulmonary bypass. METHODS: After surgery patients were randomly assigned to intensive insulin therapy (blood glucose between 80 and 110 mg/dl) or conventional insulin therapy (blood glucose less than 200 mg/dl). At 0, 1, 2, 4, 8, 12, 16 and 24 hours after admission to the intensive care unit, plasma samples and samples from the mediastinal drains were obtained. We measured the concentrations of the pro-inflammatory cytokines TNF-alpha and IL-6 and the anti-inflammatory cytokine IL-10 by enzyme-linked immunosorbent assay. RESULTS: Both patient groups were comparable in demographics, clinical characteristics and peri-operative data. In the intensive treatment group, glucose levels were significantly lower than in the conventionally treated group. No differences were found between both groups in the concentrations of TNF-alpha, IL-6 and IL-10 in plasma samples or in fluid draining the mediastinal cavity. Levels of IL-6 and IL-10 were significantly higher in mediastinal fluid samples than in plasma samples, suggesting a compartmentalized production of cytokines. CONCLUSION: The protective effect of intensive insulin therapy in patients after cardiac surgery with cardiopulmonary bypass is not related to a change in cytokine balance from a pro-inflammatory to an anti-inflammatory pattern. Systemic cytokine levels are not representative of the local inflammatory response. PMID- 16356229 TI - Does taking endurance into account improve the prediction of weaning outcome in mechanically ventilated children? AB - INTRODUCTION: We conducted the present study to determine whether a combination of the mechanical ventilation weaning predictors proposed by the collective Task Force of the American College of Chest Physicians (TF) and weaning endurance indices enhance prediction of weaning success. METHOD: Conducted in a tertiary paediatric intensive care unit at a university hospital, this prospective study included 54 children receiving mechanical ventilation (> or = 6 hours) who underwent 57 episodes of weaning. We calculated the indices proposed by the TF (spontaneous respiratory rate, paediatric rapid shallow breathing, rapid shallow breathing occlusion pressure [ROP] and maximal inspiratory pressure during an occlusion test [Pimax]) and weaning endurance indices (pressure-time index, tension-time index obtained from P(0.1) [TTI1] and from airway pressure [TTI2]) during spontaneous breathing. Performances of each TF index and combinations of them were calculated, and the best single index and combination were identified. Weaning endurance parameters (TTI1 and TTI2) were calculated and the best index was determined using a logistic regression model. Regression coefficients were estimated using the maximum likelihood ratio (LR) method. Hosmer-Lemeshow test was used to estimate goodness-of-fit of the model. An equation was constructed to predict weaning success. Finally, we calculated the performances of combinations of best TF indices and best endurance index. RESULTS: The best single TF index was ROP, the best TF combination was represented by the expression (0.66 x ROP) + (0.34 x Pimax), and the best endurance index was the TTI2, although their performance was poor. The best model resulting from the combination of these indices was defined by the following expression: (0.6 x ROP) - (0.1 x Pimax) + (0.5 x TTI2). This integrated index was a good weaning predictor (P < 0.01), with a LR+ of 6.4 and LR+/LR- ratio of 12.5. However, at a threshold value < 1.3 it was only predictive of weaning success (LR- = 0.5). CONCLUSION: The proposed combined index, incorporating endurance, was of modest value in predicting weaning outcome. This is the first report of the value of endurance parameters in predicting weaning success in children. Currently, clinical judgement associated with spontaneous breathing trials apparently remain superior. PMID- 16356230 TI - Long term effect of a medical emergency team on cardiac arrests in a teaching hospital. AB - INTRODUCTION: It is unknown whether the reported short-term reduction in cardiac arrests associated with the introduction of the medical emergency team (MET) system can be sustained. METHOD: We conducted a prospective, controlled before and-after examination of the effect of a MET system on the long-term incidence of cardiac arrests. We included consecutive patients admitted during three study periods: before the introduction of the MET; during the education phase preceding the implementation of the MET; and a period of four years from the implementation of the MET system. Cardiac arrests were identified from a log book of cardiac arrest calls and cross-referenced with case report forms and the intensive care unit admissions database. We measured the number of hospital admissions and MET reviews during each period, performed multivariate logistic regression analysis to identify predictors of mortality following cardiac arrest and studied the correlation between the rate of MET calls with the rate of cardiac arrests. RESULTS: Before the introduction of the MET system there were 66 cardiac arrests and 16,246 admissions (4.06 cardiac arrests per 1,000 admissions). During the education period, the incidence of cardiac arrests decreased to 2.45 per 1,000 admissions (odds ratio (OR) for cardiac arrest 0.60; 95% confidence interval (CI) 0.43-0.86; p = 0.004). After the implementation of the MET system, the incidence of cardiac arrests further decreased to 1.90 per 1,000 admissions (OR for cardiac arrest 0.47; 95% CI 0.35-0.62; p < 0.0001). There was an inverse correlation between the number of MET calls in each calendar year and the number of cardiac arrests for the same year (r2 = 0.84; p = 0.01), with 17 MET calls being associated with one less cardiac arrest. Male gender (OR 2.88; 95% CI 1.34-6.19) and an initial rhythm of either asystole (OR 7.58; 95% CI 3.15-18.25; p < 0.0001) or pulseless electrical activity (OR 4.09; 95% CI 1.59-10.51; p = 0.003) predicted an increased risk of death. CONCLUSION: Introduction of a MET system into a teaching hospital was associated with a sustained and progressive reduction in cardiac arrests over a four year period. Our findings show sustainability and suggest that, for every 17 MET calls, one cardiac arrest might be prevented. PMID- 16356231 TI - Mid-regional pro-adrenomedullin as a prognostic marker in sepsis: an observational study. AB - INTRODUCTION: Measurement of biomarkers is a potential approach to early assessment and prediction of mortality in patients with sepsis. The aim of the present study was to evaluate the prognostic value of mid-regional pro adrenomedullin (MR-proADM) levels in a cohort of medical intensive care patients and to compare it with other biomarkers and physiological scores. METHOD: We evaluated blood samples from 101 consecutive critically ill patients admitted to the intensive care unit and from 160 age-matched healthy control individuals. The patients had initially been enrolled in a prospective observational study investigating the prognostic value of endocrine dysfunction in critically ill patients ("PEDCRIP" Study). The prognostic value of MR-proADM levels was compared with those of two physiological scores and of various biomarkers (for example C reactive Protein, IL-6, procalcitonin). MR-proADM was measured in EDTA plasma from all patients using a new sandwich immunoassay. RESULTS: On admission, 53 patients had sepsis, severe sepsis, or septic shock, and 48 had systemic inflammatory response syndrome. Median MR-proADM levels on admission (nmol/l [range]) were 1.1 (0.3-3.7) in patients with systemic inflammatory response syndrome, 1.8 (0.4-5.8) in those with sepsis, 2.3 (1.0-17.6) in those with severe sepsis and 4.5 (0.9-21) in patients with septic shock. In healthy control individuals the median MR-proADM was 0.4 (0.21-0.97). On admission, circulating MR-proADM levels in patients with sepsis, severe sepsis, or septic shock were significantly higher in nonsurvivors (8.5 [0.8-21.0]; P < 0.001) than in survivors (1.7 [0.4-17.6]). In a receiver operating curve analysis of survival of patients with sepsis, the area under the curve (AUC) for MR-proADM was 0.81, which was similar to the AUCs for IL-6, Acute Physiology and Chronic Health Evaluation II score and Simplified Acute Physiology Score II. The prognostic value of MR-proADM was independent of the sepsis classification system used. CONCLUSION: MR-proADM may be helpful in individual risk assessment in septic patients. PMID- 16356232 TI - Tissue engineered fetal skin constructs for pediatric burns. AB - The management of patients with partial thickness (second degree) burns is problematic due to the different treatments needed for varying depths of injury. A report recently published in The Lancet describes a novel treatment for deep second degree burns using a fetal skin construct (FSC). The authors included eight pediatric patients with small second degree burns. They showed that FSCs reduced the need for autografting of deep second degree burns, with little hypertrophy of new skin and no skin contraction. This technology is new and exciting, but in our opinion several issues must be addressed before FSCs can enter the clinical arena. All of the patients were included in the treatment group, and therefore no comparison with conventional skin substitutes was possible. There is no mention of the use of laser Doppler in any initial assessment of patients. The debridement carried out before application of the FSC is not elaborated upon, and the surface areas involved in the study were very small in most cases, which limits the relevance to patients with larger burns. The use of FSCs gives us an additional option in a range of possible treatments for this notoriously difficult-to-treat patient group. PMID- 16356233 TI - Recently published papers: treating sepsis, measuring troponin and managing the obese. AB - Sepsis and septic shock continue to contribute to our workload and stimulate our research activities although many fundamental questions remain. Studies reported on here focus on inotrope use and a novel way of predicting inotrope response. Continuing this theme more fundamental work is reported examining the mitochondrial respiratory chain and the effects of sepsis coupled with interesting work on lactic acidosis. Troponin raises its head again and we are still left quizzing over its value in the ICU. Finally we discuss a paper on the outcome of the obese patient on a general ICU. Like sepsis a continuing challenge. PMID- 16356234 TI - Pro/con ethics debate: when is dead really dead? AB - Contemporary intensive care unit (ICU) medicine has complicated the issue of what constitutes death in a life support environment. Not only is the distinction between sapient life and prolongation of vital signs blurred but the concept of death itself has been made more complex. The demand for organs to facilitate transplantation promotes a strong incentive to define clinical death in a manner that most effectively supplies that demand. We consider the problem of defining death in the ICU as a function of viable organ availability for transplantation. PMID- 16356235 TI - Bench-to-bedside review: Antidotal treatment of sulfonylurea-induced hypoglycaemia with octreotide. AB - The major potential adverse effect of use of sulfonylurea agents (SUAs) is a hyperinsulinaemic state that causes hypoglycaemia. It may be observed during chronic therapeutic dosing, even with very low doses of a SUA, and especially in older patients. It may also result from accidental or intentional poisoning in both diabetic and nondiabetic patients. The traditional approach to SUA-induced hypoglycaemia includes administration of glucose, and glucagon or diazoxide in those who remain hypoglycaemic despite repeated or continuous glucose supplementation. However, these antidotal approaches are associated with several shortcomings, including further exacerbation of insulin release by glucose and glucagon, leading only to a temporary beneficial effect and later relapse into hypoglycaemia, as well as the adverse effects of both glucagon and diazoxide. Octreotide inhibits the secretion of several neuropeptides, including insulin, and has successfully been used to control life-threatening hypoglycaemia caused by insulinoma or persistent hyperinsulinaemic hypoglycaemia of infancy. Therefore, this agent should in theory also be useful to decrease glucose requirements and the number of hypoglycaemic episodes in patients with SUA induced hypoglycaemia. This has apparently been confirmed by experimental data, one retrospective study based on chart review, and several anecdotal case reports. There is thus a need for further prospective studies, which should be adequately powered, randomized and controlled, to confirm the probable beneficial effect of octreotide in this setting. PMID- 16356236 TI - Bench-to-bedside review: Paediatric viral lower respiratory tract disease necessitating mechanical ventilation--should we use exogenous surfactant? AB - Treatment of infants with viral lower respiratory tract disease (LRTD) necessitating mechanical ventilation is mainly symptomatic. The therapeutic use of surfactant seems rational because significantly lower levels of surfactant phospholipids and proteins, and impaired capacity to reduce surface tension were observed among infants and young children with viral LRTD. This article reviews the role of pulmonary surfactant in the pathogenesis of paediatric viral LRTD. Three randomized trials demonstrated improved oxygenation and reduced duration of mechanical ventilation and paediatric intensive care unit stay in young children with viral LRTD after administration of exogenous surfactant. This suggest that exogenous surfactant is the first beneficial treatment for ventilated infants with viral LRTD. Additionally, in vitro and animal studies demonstrated that surfactant associated proteins SP-A and SP-D bind to respiratory viruses, play a role in eliminating these viruses and induce an inflammatory response. Although these immunomodulating effects are promising, the available data are inconclusive and the findings are unconfirmed in humans. In summary, exogenous surfactant in ventilated infants with viral LRTD could be a useful therapeutic approach. Its beneficial role in improving oxygenation has already been established in clinical trials, whereas the immunomodulating effects are promising but remain to be elucidated. PMID- 16356237 TI - Bench-to-bedside review: Amelioration of acute renal impairment using ethyl pyruvate. AB - Inflammation and oxidative stress cause renal impairment. Renal failure exacerbates the effect of oxidative stress on many organ systems. Antioxidants can prevent or treat renal failure in various experimental models and clinical situations. Pyruvate is an endogenous antioxidant with beneficial effects in animal models of oxidative stress. Because sodium pyruvate rapidly degrades in solution, a simple derivative of pyruvic acid, namely ethyl pyruvate, has been investigated as a therapeutic agent in preclinical studies. Ethyl pyruvate reduces organ system damage in ischaemia/reperfusion injury and haemorrhagic and endotoxic shock, at least in part through its antioxidant action. In addition, ethyl pyruvate appears to have direct beneficial effects on cytokine expression and proinflammatory gene regulation. The effect is long lasting and, importantly, even when it is administered after the onset of inflammation it can ameliorate organ damage and improve survival. Ethyl pyruvate is a widely used as a food additive and was shown to be safe in phase I clinical trials. We suggest ethyl pyruvate warrants further evaluation in the management of acute renal impairment. PMID- 16356239 TI - Bench-to-bedside review: Is there a place for epinephrine in septic shock? AB - The use of epinephrine in septic shock remains controversial. Nevertheless, epinephrine is widely used around the world and the reported morbidity and mortality rates with it are no different from those observed with other vasopressors. In volunteers, epinephrine increases heart rate, mean arterial pressure and cardiac output. Epinephrine also induces hyperglycemia and hyperlactatemia. In hyperkinetic septic shock, epinephrine consistently increases arterial pressure and cardiac output in a dose dependent manner. Epinephrine transiently increases lactate levels through an increase in aerobic glycolysis. Epinephrine has no effect on splanchnic circulation in dopamine-sensitive septic shock. On the other hand, in dopamine-resistant septic shock, epinephrine has no effect on tonometric parameters but decreases fractional splanchnic blood flow with an increase in the gradient of mixed venous oxygen saturation (SVO2) and hepatic venous oxygen saturation (SHO2). In conclusion, epinephrine has predictable effects on systemic hemodynamics and is as efficient as norepinephrine in correcting hemodynamic disturbances of septic shock. Moreover, epinephrine is cheaper than other commonly used catecholamine regimens in septic shock. The clinical impact of the transient hyperlactatemia and of the splanchnic effects are not established. PMID- 16356240 TI - Functional hemodynamic monitoring. AB - Hemodynamic monitoring is a central component of intensive care. Patterns of hemodynamic variables often suggest cardiogenic, hypovolemic, obstructive, or distributive (septic) etiologies to cardiovascular insufficiency, thus defining the specific treatments required. Monitoring increases in invasiveness, as required, as the risk for cardiovascular instability-induced morbidity increases because of the need to define more accurately the diagnosis and monitor the response to therapy. Monitoring is also context specific: requirements during cardiac surgery will be different from those in the intensive care unit or emergency department. Solitary hemodynamic values are useful as threshold monitors (e.g. hypotension is always pathological, central venous pressure is only elevated in disease). Some hemodynamic values can only be interpreted relative to metabolic demand, whereas others have multiple meanings. Functional hemodynamic monitoring implies a therapeutic application, independent of diagnosis such as a therapeutic trial of fluid challenge to assess preload responsiveness. Newer methods for assessing preload responsiveness include monitoring changes in central venous pressure during spontaneous inspiration, and variations in arterial pulse pressure, systolic pressure, and aortic flow variation in response to vena caval collapse during positive pressure ventilation or passive leg raising. Defining preload responsiveness using these functional measures, coupled to treatment protocols, can improve outcome from critical illness. Potentially, as these and newer, less invasive hemodynamic measures are validated, they could be incorporated into such protocolized care in a cost effective manner. PMID- 16356241 TI - Clinical review: Renal tubular acidosis--a physicochemical approach. AB - The Canadian physiologist PA Stewart advanced the theory that the proton concentration, and hence pH, in any compartment is dependent on the charges of fully ionized and partly ionized species, and on the prevailing CO2 tension, all of which he dubbed independent variables. Because the kidneys regulate the concentrations of the most important fully ionized species ([K+], [Na+], and [Cl ]) but neither CO2 nor weak acids, the implication is that it should be possible to ascertain the renal contribution to acid-base homeostasis based on the excretion of these ions. One further corollary of Stewart's theory is that, because pH is solely dependent on the named independent variables, transport of protons to and from a compartment by itself will not influence pH. This is apparently in great contrast to models of proton pumps and bicarbonate transporters currently being examined in great molecular detail. Failure of these pumps and cotransporters is at the root of disorders called renal tubular acidoses. The unquestionable relation between malfunction of proton transporters and renal tubular acidosis represents a problem for Stewart theory. This review shows that the dilemma for Stewart theory is only apparent because transport of acid-base equivalents is accompanied by electrolytes. We suggest that Stewart theory may lead to new questions that must be investigated experimentally. Also, recent evidence from physiology that pH may not regulate acid-base transport is in accordance with the concepts presented by Stewart. PMID- 16356242 TI - Clinical review: Mechanical ventilation in severe asthma. AB - Respiratory failure from severe asthma is a potentially reversible, life threatening condition. Poor outcome in this setting is frequently a result of the development of gas-trapping. This condition can arise in any mechanically ventilated patient, but those with severe airflow limitation have a predisposition. It is important that clinicians managing these types of patients understand that the use of mechanical ventilation can lead to or worsen gas trapping. In this review we discuss the development of this complication during mechanical ventilation, techniques to measure it and strategies to limit its severity. We hope that by understanding such concepts clinicians will be able to reduce further the poor outcomes occasionally related to severe asthma. PMID- 16356243 TI - Lactate as a marker of energy failure in critically ill patients: hypothesis. AB - Lactate measurement in the critically ill has been traditionally used to stratify patients with poor outcome. However, plasma lactate levels are the result of a finely tuned interplay of factors that affect the balance between its production and its clearance. When the oxygen supply does not match its consumption, organisms such as man who are forced to produce ATP for their integrity adapt in many different ways up to the point when energy failure occurs. Lactate, being part of the adaptive response, may then be used to assess the severity of the supply/demand imbalance. In such a scenario, the time to intervention becomes relevant: early and effective treatment may allow the cell to revert to a normal state, as long as the oxygen machinery (i.e. mitochondria) is intact. Conversely, once the mitochondria are deranged, energy failure occurs even in the presence of normoxia. The lactate increase in critically ill patients may therefore be viewed as an early marker of a potentially reversible state. PMID- 16356244 TI - Clinical review: Independent lung ventilation in critical care. AB - Independent lung ventilation (ILV) can be classified into anatomical and physiological lung separation. It requires either endobronchial blockade or double-lumen endotracheal tube intubation. Endobronchial blockade or selective double-lumen tube ventilation may necessitate temporary one lung ventilation. Anatomical lung separation isolates a diseased lung from contaminating the non diseased lung. Physiological lung separation ventilates each lung as an independent unit. There are some clear indications for ILV as a primary intervention and as a rescue ventilator strategy in both anatomical and physiological lung separation. Potential pitfalls are related to establishing and maintaining lung isolation. Nevertheless, ILV can be used in the intensive care setting safely with a good understanding of its limitations and potential complications. PMID- 16356245 TI - Clinical review: interpretation of arterial pressure wave in shock states. AB - In critically ill patients monitored with an arterial catheter, the arterial pressure signal provides two types of information that may help the clinician to interpret haemodynamic status better: the mean values of systolic, diastolic, mean and pulse pressures; and the magnitude of the respiratory variation in arterial pressure in patients undergoing mechanical ventilation. In this review we briefly discuss the physiological mechanisms responsible for arterial pressure generation, with special focus on resistance, compliance and pulse wave amplification phenomena. We also emphasize the utility of taking into consideration the overall arterial pressure set (systolic, diastolic, mean and pulse pressures) in order to define haemodynamic status better. Finally, we review recent studies showing that quantification of respiratory variation in pulse and systolic arterial pressures can allow one to identify the mechanically ventilated patients who may benefit from volume resuscitation. PMID- 16356247 TI - Lactate in the intensive care unit: pyromaniac, sentinel or fireman? AB - Lactate, indispensable substrate of mammalian intermediary metabolism, allows shuttling of carbons and reducing power between cells and organs at a high turnover rate. Lactate is, therefore, not deleterious, although an increase in its concentration is often a sensitive sign of alteration in energy homeostasis, a rise in it being frequently related to poor prognosis. Such an increase, however, actually signifies an attempt by the body to cope with a new energy status. Hyperlactatemia, therefore, most often represents an adaptive response to an acute energy disorder. Investigation of lactate metabolism at the bedside is limited to the determination of its concentration. Lactate metabolism and acid base homeostasis are both closely linked to cellular energy metabolism, acidosis being potentially a cause or a consequence of cellular energy deficit. PMID- 16356248 TI - Antithrombin in sepsis revisited. AB - As the pivotal phase III randomized controlled clinical trial on antithrombin concentrate in patients with severe sepsis did not show a beneficial effect of antithrombin treatment on 28-day mortality, the interest in the potential use of this treatment modality in sepsis has diminished. However, recent data on the effect of antithrombin administration on coagulation in combination with recent analyses from the clinical trials that were aimed to restore physiological anticoagulant pathways in patients with sepsis may revitalize the interest in antithrombin concentrate for the treatment of severe sepsis. PMID- 16356246 TI - Clinical review: Positive end-expiratory pressure and cardiac output. AB - In patients with acute lung injury, high levels of positive end-expiratory pressure (PEEP) may be necessary to maintain or restore oxygenation, despite the fact that 'aggressive' mechanical ventilation can markedly affect cardiac function in a complex and often unpredictable fashion. As heart rate usually does not change with PEEP, the entire fall in cardiac output is a consequence of a reduction in left ventricular stroke volume (SV). PEEP-induced changes in cardiac output are analyzed, therefore, in terms of changes in SV and its determinants (preload, afterload, contractility and ventricular compliance). Mechanical ventilation with PEEP, like any other active or passive ventilatory maneuver, primarily affects cardiac function by changing lung volume and intrathoracic pressure. In order to describe the direct cardiocirculatory consequences of respiratory failure necessitating mechanical ventilation and PEEP, this review will focus on the effects of changes in lung volume, factors controlling venous return, the diastolic interactions between the ventricles and the effects of intrathoracic pressure on cardiac function, specifically left ventricular function. Finally, the hemodynamic consequences of PEEP in patients with heart failure, chronic obstructive pulmonary disease and acute respiratory distress syndrome are discussed. PMID- 16356249 TI - The staging of sepsis: understanding heterogeneity in treatment efficacy. AB - Human sepsis is an intrinsically complex disease. Populations of patients enrolled into clinical trials of novel sepsis therapies are notoriously heterogeneous with respect to the inciting cause of their disease, the co-morbid conditions that define its course, and the acute severity of their initial presentation. This heterogeneity is reflected in strikingly variable mortality risks across studies, and probably, though less clearly-established, in variable response rates to a given intervention. In an accompanying article in this issue of Critical Care, Macias and colleagues argue that the effectiveness of adjuvant sepsis therapies is not dependent on the baseline mortality risk, since the few "positive" trials that have been published show widely divergent placebo mortality rates. But this analysis assumes that biologically distinct interventions will be equally efficacious in clinically diverse populations, and confuses severity as a population descriptor with severity as a surrogate measure of a biologic state in an individual patient. In a pivotal trial of recombinant human activated protein C (rhAPC) in patients with severe sepsis, an aggregate 6% mortality decrement appeared to be the result of negligible efficacy in the least severely ill patients, and considerably greater efficacy in those who were at greatest risk of dying. A larger follow-up study recruiting low risk patients confirmed this impression, showing a convincing absence of benefit in patients who clinicians judged to be less severely ill. If we accept Macias' argument, we are led to the conclusion that rhAPC is of limited use in the management of severe sepsis. On the other hand, if we view severity as a crude surrogate for a particular pathologic state, we would shift our focus to better defining those populations most likely to benefit from intervention, including patients who may not have met criteria for entry in the original PROWESS trial--those with disseminated intravascular coagulation or acute organ dysfunction from causes other than sepsis. Staging systems that stratify heterogeneous patient populations by risk and by potential to benefit from intervention have proven to be essential to the development of multimodal adjuvant treatment for cancer. They will be no less important in the optimal management of sepsis. PMID- 16356250 TI - Open the intensive care unit doors to HIV-infected patients with sepsis. AB - Severe sepsis is more and more frequent, especially because of an increased rate of immunocompromised patients. Despite the improvement in the overall prognosis of HIV/AIDS patients and the improvement of global ICU care, the prognosis of HIV/ADS patients hospitalized in ICU with severe sepsis remained poor. This situation is partly due to the increased proportion of HIV/AIDS patients with limited access to health care and to a reluctance of ICU physicians in admitting HIV infected patients. However, medical literature suggests that ICU prognosis of immunocompromised (especially cancer) patients should be largely improved by early ICU admission and by an early institution of supportive techniques. This strategy should be used in HIV/AIDS patients with severe sepsis. PMID- 16356251 TI - The place of early haemoperfusion with polymyxin B fibre column in the treatment of sepsis. AB - Direct haemoperfusion with polymyxin B-immobilized fibre (PMX-F) is a promising treatment for Gram-negative sepsis in critically ill patients. Indeed, it has been used routinely in Japan for a decade. Recent evidence presented in this journal suggests that PMX-F can have a positive impact on outcome in patients with sepsis, although other reports in the literature have presented confusing or even conflicting results. This commentary considers whether the available evidence allows us to establish an appropriate role for PMX-F treatment in sepsis and what further work is needed. PMID- 16356252 TI - Myocardial infarction complicating critical illness. AB - Cardiac troponins are highly sensitive and specific indicators of myocardial injury. Although the mechanism of this injury is not entirely clear, it carries important prognostic information. Elevated serum levels of cardiac troponins have been described in a wide variety of conditions other than myocardial infarction (MI). The current study is an important first step in trying to determine the exact frequency of MI among critically ill patients with elevated troponin. At present, the rate of MI in these patients is unknown and its implications on outcome and management will have to await future prospective clinical trials. PMID- 16356253 TI - Predictive mortality models are not like fine wine. AB - The authors of a recent paper have described an updated simplified acute physiology score (SAPS) II mortality model developed on patient data from 1998 to 1999. Hospital mortality models have a limited range of applicability. SAPS II, Acute Physiology, Age, and Chronic Health Evaluation (APACHE) III, and mortality probability model (MPM)-II, which were developed in the early 1990s, have shown a decline in predictive accuracy as the models age. The deterioration in accuracy is manifested by a decline in the models' calibration. In particular, mortality tends to get over predicted when older models are applied to more contemporary data, which in turn leads to 'grade inflation' when benchmarking intensive care unit (ICU) performance. Although the authors claim that their updated SAPS II can be used for benchmarking ICU performance, it seems likely that this model might already be out of calibration for patient data collected in 2005 and beyond. Thus, the updated SAPS II model may be interesting for historical purposes, but it is doubtful that it can be an accurate tool for benchmarking data from contemporary populations. PMID- 16356254 TI - Cerebral perfusion pressure and brain ischaemia: can one size fit all? AB - Current recommendations regarding the management of patients after traumatic brain injury include reduction in brain tissue pressure (i.e. intracranial pressure) and maintenance of an adequate arterial pressure; these measures combined should result in cerebral perfusion pressure sufficient to achieve adequate oxygen delivery. After almost 20 years of observational studies comparing cerebral perfusion pressure and indices of cerebral oxygenation, it is apparent that there is no single value for cerebral perfusion pressure that, if achieved, will provide adequate cerebral oxygen delivery in all patients. Traumatic brain injury remains a common problem, and this should encourage researchers and clinicians to design better and adequately powered trials of monitors and associated interventions. PMID- 16356255 TI - Can we predict septic shock in patients with hospital-acquired pneumonia? AB - Hospital-acquired pneumonia is a serious and potentially life-threatening complication, with reported pneumonia-attributable mortality rates as high as 50%. Rapid diagnosis and immediate institution of adequate empirical antimicrobial treatment are of paramount importance in patient management. Nevertheless, some patients deteriorate and develop respiratory insufficiency, septic shock and a multiorgan dysfunction syndrome. Early recognition of these patients might help in reducing morbidity and mortality. Elevated systemic levels of proinflammatory cytokines (IL-1beta, IL-6, IL-8 and IL-10) at the time of diagnosis of hospital-acquired pneumonia appear to be indicative of subsequent progression to septic shock. Should this now become a part of patient management? PMID- 16356256 TI - Highs and lows in high-risk surgery: the controversy of goal-directed haemodynamic management. AB - Although various systems have been developed to identify patients at increased risk of peri- and postoperative mortality and morbidity, little effort has been made in developing tools to reduce this risk. In this issue of Critical Care, Pearse et al. publish two reports related to predicting and improving outcome in high-risk surgical patients. Rather than conducting large, multicentre, randomised, controlled trials, the research group at St George's Hospital in London has persistently and systematically tested the concept of goal-directed haemodynamic management in high risk surgery in their single-centre setting. Their results have been impressive, demonstrating that in this setting, various outcome measures can be reduced with goal-directed haemodynamic management. The impressive positive results of the Pearse studies contrast sharply with the negative results of multicentre studies, such as that of Sandham et al. One reason may be that, like several other successful single-centre trials, Pearse et al. used strict treatment protocols rather than guidelines. In addition, single centre studies utilize their investigators' knowledge of their patients' risk profiles and familiarity with the care processes and infrastructures of their institutions. An understanding of the organisational and case-mix aspects of pre , peri-and post-operative management is vital for planning multicentre trials of goal-directed management. PMID- 16356257 TI - Optimal management of the high risk surgical patient: beta stimulation or beta blockade? AB - Several groups of investigators have shown that peri-operative goal directed therapy (GDT) may reduce mortality in high-risk surgical patients. GDT usually requires the use of beta-adrenergic agents, however, and these may also carry the risk of cardiac ischemia, especially in patients with ischemic diseases. In this commentary, we will discuss the apparent contradiction between studies showing beneficial effects of GDT in high-risk surgical patients and studies showing the benefit of beta-blockade in high-risk surgery. One of the key differences between both types of studies is that GDT is applied in patients with high risk of postoperative death, excluding patients with cardiac ischemic disease, while studies reporting beneficial effects of beta-blockade have investigated patients with high risk of cardiac ischemia but moderate risk of death related to the surgical procedure itself. It is likely that beta-blockade should be proposed in patients with moderate risk of death, whereas GDT using fluids and inotropic agents should be applied in patients with high risk of peri-operative death. Monitoring central venous oxygen saturation may be useful to individualize therapy, but further studies are required to validate this option. PMID- 16356258 TI - Goal directed therapy: how long can we wait? AB - Intensive monitoring and aggressive management of perioperative haemodynamics (goal directed therapy) have repeatedly been reported to reduce the significant morbidity and mortality associated with high risk surgery. It may not matter what particular monitor is used to assess cardiac output but it is essential to ensure adequate oxygen delivery. If this management cannot begin preoperatively, it is still worth beginning goal directed therapy in the immediate postoperative period. PMID- 16356259 TI - Conflicting clinical trial data: a lesson from albumin. AB - Albumin is a frequently prescribed drug in hospitalized patients, and its effect on clinical outcomes has been scrutinized in recent years. Data from meta analyses has suggested harm related to albumin therapy in critically ill patients, and new observational data are consistent with these results. However, appropriately powered randomized, controlled trials have shown albumin to be safe in broad groups of critically ill patients. This article will discuss the reasons for differences between observational and controlled trial data, and the implications for future albumin use and clinical research. PMID- 16356260 TI - Prediction of ventilation weaning outcome: children are not little adults. AB - Prediction of ventilation weaning outcome in children is important, as unsuccessful extubation increases both morbidity and mortality. Adult weaning criteria are poor predictors of weaning outcome in children for several possible reasons: the length of mechanical ventilation is generally much shorter, and the weaning failure rate is lower in children (thus larger patient numbers are required); integrated weaning indices, such as the rapid shallow breathing index, do not account for normal developmental changes in respiratory function; and the heterogeneity of mechanically ventilated children is greater than in adults. The challenge remains to find universal weaning outcome predictors in children. PMID- 16356261 TI - The Surviving Sepsis Campaign sepsis change bundles and clinical practice. AB - The Surviving Sepsis Campaign (SSC) is an international effort to reduce mortality in severe sepsis and septic shock. The campaign included the creation of evidence-based guidelines sponsored and endorsed by 11 international organizations. From these guidelines, sepsis change bundles for initial resuscitation (6 hours) and management (24 hours) were created as a performance improvement tool. In this issue of Critical Care, Gao et al. have evaluated performance at their institution by using a close adaptation of the two SSC bundle sets and demonstrated an association between 100% compliance with the bundle elements and clinical outcome. The next step will be to demonstrate that the use of education and feedback for performance improvement will increase compliance and decrease mortality in the patient population in general. PMID- 16356262 TI - Penetration of the English language in science: the case of a German national interdisciplinary critical care conference. PMID- 16356263 TI - Biased alternative polyadenylation in human tissues. AB - BACKGROUND: Alternative polyadenylation is one of the mechanisms in human cells that give rise to a variety of transcripts from a single gene. More than half of the human genes have multiple polyadenylation sites (poly(A) sites), leading to variable mRNA and protein products. Previous studies of individual genes have indicated that alternative polyadenylation could occur in a tissue-specific manner. RESULTS: We set out to systematically investigate the occurrence and mechanism of alternative polyadenylation in different human tissues using bioinformatic approaches. Using expressed sequence tag (EST) data, we investigated 42 distinct tissue types. We found that several tissues tend to use poly(A) sites that are biased toward certain locations of a gene, such as sites located in introns or internal exons, and various sites in the exon located closest to the 3' end. We also identified several tissues, including eye, retina and placenta, that tend to use poly(A) sites not frequently used in other tissues. By exploring microarray expression data, we analyzed over 20 genes whose protein products are involved in the process or regulation of mRNA polyadenylation. Several brain tissues showed high concordance of gene expression of these genes with each other, but low concordance with other tissue types. By comparing genomic regions surrounding poly(A) sites preferentially used in brain tissues with those in other tissues, we identified several cis-regulatory elements that were significantly associated with brain-specific poly(A) sites. CONCLUSION: Our results indicate that there are systematic differences in poly(A) site usage among human tissues, and both trans-acting factors and cis-regulatory elements may be involved in regulating alternative polyadenylation in different tissues. PMID- 16356264 TI - Gene expression and metabolite profiling of Populus euphratica growing in the Negev desert. AB - BACKGROUND: Plants growing in their natural habitat represent a valuable resource for elucidating mechanisms of acclimation to environmental constraints. Populus euphratica is a salt-tolerant tree species growing in saline semi-arid areas. To identify genes involved in abiotic stress responses under natural conditions we constructed several normalized and subtracted cDNA libraries from control, stress exposed and desert-grown P. euphratica trees. In addition, we identified several metabolites in desert-grown P. euphratica trees. RESULTS: About 14,000 expressed sequence tag (EST) sequences were obtained with a good representation of genes putatively involved in resistance and tolerance to salt and other abiotic stresses. A P. euphratica DNA microarray with a uni-gene set of ESTs representing approximately 6,340 different genes was constructed. The microarray was used to study gene expression in adult P. euphratica trees growing in the desert canyon of Ein Avdat in Israel. In parallel, 22 selected metabolites were profiled in the same trees. CONCLUSION: Of the obtained ESTs, 98% were found in the sequenced P. trichocarpa genome and 74% in other Populus EST collections. This implies that the P. euphratica genome does not contain different genes per se, but that regulation of gene expression might be different and that P. euphratica expresses a different set of genes that contribute to adaptation to saline growth conditions. Also, all of the five measured amino acids show increased levels in trees growing in the more saline soil. PMID- 16356265 TI - Identification of novel Y chromosome encoded transcripts by testis transcriptome analysis of mice with deletions of the Y chromosome long arm. AB - BACKGROUND: The male-specific region of the mouse Y chromosome long arm (MSYq) is comprised largely of repeated DNA, including multiple copies of the spermatid expressed Ssty gene family. Large deletions of MSYq are associated with sperm head defects for which Ssty deficiency has been presumed to be responsible. RESULTS: In a search for further candidate genes associated with these defects we analyzed changes in the testis transcriptome resulting from MSYq deletions, using testis cDNA microarrays. This approach, aided by accumulating mouse MSYq sequence information, identified transcripts derived from two further spermatid-expressed multicopy MSYq gene families; like Ssty, each of these new MSYq gene families has multicopy relatives on the X chromosome. The Sly family encodes a protein with homology to the chromatin-associated proteins XLR and XMR that are encoded by the X chromosomal relatives. The second MSYq gene family was identified because the transcripts hybridized to a microarrayed X chromosome-encoded testis cDNA. The X loci ('Astx') encoding this cDNA had 92-94% sequence identity to over 100 putative Y loci ('Asty') across exons and introns; only low level Asty transcription was detected. More strongly transcribed recombinant loci were identified that included Asty exons 2-4 preceded by Ssty1 exons 1, 2 and part of exon 3. Transcription from the Ssty1 promotor generated spermatid-specific transcripts that, in addition to the variable inclusion of Ssty1 and Asty exons, included additional exons because of the serendipitous presence of splice sites further downstream. CONCLUSION: We identified further MSYq-encoded transcripts expressed in spermatids and deriving from multicopy Y genes, deficiency of which may underlie the defects in sperm development associated with MSYq deletions. PMID- 16356266 TI - The design of transcription-factor binding sites is affected by combinatorial regulation. AB - BACKGROUND: Transcription factors regulate gene expression by binding to specific cis-regulatory elements in gene promoters. Although DNA sequences that serve as transcription-factor binding sites have been characterized and associated with the regulation of numerous genes, the principles that govern the design and evolution of such sites are poorly understood. RESULTS: Using the comprehensive mapping of binding-site locations available in Saccharomyces cerevisiae, we examined possible factors that may have an impact on binding-site design. We found that binding sites tend to be shorter and fuzzier when they appear in promoter regions that bind multiple transcription factors. We further found that essential genes bind relatively fewer transcription factors, as do divergent promoters. We provide evidence that novel binding sites tend to appear in specific promoters that are already associated with multiple sites. CONCLUSION: Two principal models may account for the observed correlations. First, it may be that the interaction between multiple factors compensates for the decreased specificity of each specific binding sequence. In such a scenario, binding-site fuzziness is a consequence of the presence of multiple binding sites. Second, binding sites may tend to appear in promoter regions that are subject to low selective pressure, which also allows for fuzzier motifs. The latter possibility may account for the relatively low number of binding sites found in promoters of essential genes and in divergent promoters. PMID- 16356267 TI - The discovery, positioning and verification of a set of transcription-associated motifs in vertebrates. AB - We have developed several new methods to investigate transcriptional motifs in vertebrates. We developed a specific alignment tool appropriate for regions involved in transcription control, and exhaustively enumerated all possible 12 mers for involvement in transcription by virtue of their mammalian conservation. We then used deeper comparative analysis across vertebrates to identify the active instances of these motifs. We have shown experimentally in Medaka fish that a subset of these predictions is involved in transcription. PMID- 16356268 TI - A low-cost open-source SNP genotyping platform for association mapping applications. AB - Association mapping aimed at identifying DNA polymorphisms that contribute to variation in complex traits entails genotyping a large number of single nucleotide polymorphisms (SNPs) in a very large panel of individuals. Few technologies, however, provide inexpensive high-throughput genotyping. Here, we present an efficient approach developed specifically for genotyping large fixed panels of diploid individuals. The cost-effective, open-source nature of our methodology may make it particularly attractive to those working in nonmodel systems. PMID- 16356269 TI - Ulysses - an application for the projection of molecular interactions across species. AB - We developed Ulysses as a user-oriented system that uses a process called Interolog Analysis for the parallel analysis and display of protein interactions detected in various species. Ulysses was designed to perform such Interolog Analysis by the projection of model organism interaction data onto homologous human proteins, and thus serves as an accelerator for the analysis of uncharacterized human proteins. The relevance of projections was assessed and validated against published reference collections. All source code is freely available, and the Ulysses system can be accessed via a web interface http://www.cisreg.ca/ulysses. PMID- 16356270 TI - Protein-protein interactions of the hyperthermophilic archaeon Pyrococcus horikoshii OT3. AB - BACKGROUND: Although 2,061 proteins of Pyrococcus horikoshii OT3, a hyperthermophilic archaeon, have been predicted from the recently completed genome sequence, the majority of proteins show no similarity to those from other organisms and are thus hypothetical proteins of unknown function. Because most proteins operate as parts of complexes to regulate biological processes, we systematically analyzed protein-protein interactions in Pyrococcus using the mammalian two-hybrid system to determine the function of the hypothetical proteins. RESULTS: We examined 960 soluble proteins from Pyrococcus and selected 107 interactions based on luciferase reporter activity, which was then evaluated using a computational approach to assess the reliability of the interactions. We also analyzed the expression of the assay samples by western blot, and a few interactions by in vitro pull-down assays. We identified 11 hetero-interactions that we considered to be located at the same operon, as observed in Helicobacter pylori. We annotated and classified proteins in the selected interactions according to their orthologous proteins. Many enzyme proteins showed self interactions, similar to those seen in other organisms. CONCLUSION: We found 13 unannotated proteins that interacted with annotated proteins; this information is useful for predicting the functions of the hypothetical Pyrococcus proteins from the annotations of their interacting partners. Among the heterogeneous interactions, proteins were more likely to interact with proteins within the same ortholog class than with proteins of different classes. The analysis described here can provide global insights into the biological features of the protein protein interactions in P. horikoshii. PMID- 16356273 TI - Ancient genomes. AB - Ever since its invention, the polymerase chain reaction has been the method of choice for work with ancient DNA. In an application of modern genomic methods to material from the Pleistocene, a recent study has instead undertaken to clone and sequence a portion of the ancient genome of the cave bear. PMID- 16356271 TI - The genomic response to 20-hydroxyecdysone at the onset of Drosophila metamorphosis. AB - BACKGROUND: The steroid hormone 20-hydroxyecdysone (20E) triggers the major developmental transitions in Drosophila, including molting and metamorphosis, and provides a model system for defining the developmental and molecular mechanisms of steroid signaling. 20E acts via a heterodimer of two nuclear receptors, the ecdysone receptor (EcR) and Ultraspiracle, to directly regulate target gene transcription. RESULTS: Here we identify the genomic transcriptional response to 20E as well as those genes that are dependent on EcR for their proper regulation. We show that genes regulated by 20E, and dependent on EcR, account for many transcripts that are significantly up- or downregulated at puparium formation. We provide evidence that 20E and EcR participate in the regulation of genes involved in metabolism, stress, and immunity at the onset of metamorphosis. We also present an initial characterization of a 20E primary-response regulatory gene identified in this study, brain tumor (brat), showing that brat mutations lead to defects during metamorphosis and changes in the expression of key 20E-regulated genes. CONCLUSION: This study provides a genome-wide basis for understanding how 20E and its receptor control metamorphosis, as well as a foundation for functional genomic analysis of key regulatory genes in the 20E signaling pathway during insect development. PMID- 16356274 TI - Splicing regulators: targets and drugs. AB - Silencing of splicing regulators by RNA interference, combined with splicing specific microarrays, has revealed a complex network of distinct alternative splicing events in Drosophila, while a high-throughput screen of more than 6,000 compounds has identified drugs that interfere specifically and directly with one class of splicing regulators in human cells. PMID- 16356275 TI - Predicting the sites of metastases. AB - Transplantation of human breast cancer cells into immunodeficient mice together with gene-expression microarray studies has recently identified genes implicated in the tissue tropism of breast-cancer metastasis. Such signatures of site specific metastatic capabilities might allow the targeting of therapy to likely sites of metastasis. PMID- 16356277 TI - Understanding embryonic development: from screens to genes. AB - A report on the 64th Annual Meeting of the Society for Developmental Biology, San Francisco, USA, 27 July-1 August 2005. PMID- 16356276 TI - The expansin superfamily. AB - The expansin superfamily of plant proteins is made up of four families, designated alpha-expansin, beta-expansin, expansin-like A and expansin-like B. alpha-Expansin and beta-expansin proteins are known to have cell-wall loosening activity and to be involved in cell expansion and other developmental events during which cell-wall modification occurs. Proteins in these two families bind tightly to the cell wall and their activity is typically assayed by their stimulation of cell-wall extension and stress relaxation; no bona fide enzymatic activity has been detected for these proteins. Alpha-expansin proteins and some, but not all, beta-expansin proteins are implicated as catalysts of 'acid growth', the enlargement of plant cells stimulated by low extracellular pH. A divergent group of beta-expansin genes are expressed at high levels in the pollen of grasses but not of other plant groups. They probably function to loosen maternal cell walls during growth of the pollen tube towards the ovary. All expansins consist of two domains; domain 1 is homologous to the catalytic domain of proteins in the glycoside hydrolase family 45 (GH45); expansin domain 2 is homologous to group-2 grass pollen allergens, which are of unknown biological function. Experimental evidence suggests that expansins loosen cell walls via a nonenzymatic mechanism that induces slippage of cellulose microfibrils in the plant cell wall. PMID- 16356278 TI - Defining the proteome. AB - A report on the Fourth Annual HUPO World Congress (HUPO2005) 'From Defining the Proteome to Understanding Function', Munich, Germany, 28 August-1 September 2005. PMID- 16356279 TI - Transposon technology and vertebrate functional genomics. AB - A report on the Third Annual International Conference on Transposition and Animal Biotechnology, Minneapolis, USA, 23-24 June 2005, and the FASEB Summer Research Conference 'Mammalian Mobile Elements', Tuscon, USA, 4-9 June, 2005. PMID- 16356282 TI - QTL-based evidence for the role of epistasis in evolution. AB - The extent to which epistasis contributes to adaptation and speciation has been a controversial topic in evolutionary genetics. One experimental approach to study epistasis is based on quantitative trait locus (QTL) mapping using molecular markers. Comparisons can be made among all possible pair-wise combinations of the markers, irrespective of whether an additive QTL is associated with a marker; several software packages have been developed that facilitate this. We review several examples of using this approach to identify epistatic QTLs for traits of evolutionary or ecological interest. While there is variability in the results, the number of epistatic QTL interactions is often greater than or equal to the number of additive QTLs. The magnitude of epistatic effects can be larger than the additive effects. Thus, epistatic interactions seem to be an important part of natural genetic variation. Future studies of epistatic QTLs could lead to descriptions of the genetic networks underlying variation for fitness-related traits. PMID- 16356283 TI - Mapping segregation distortion loci and quantitative trait loci for spikelet sterility in rice ( Oryza sativa L.). AB - Markers with segregation ratio distortion are commonly observed in data sets used for quantitative trait locus (QTL) mapping. In this study, a multipoint method of maximum likelihood (ML) was newly developed to estimate the positions and effects of the segregation distortion loci (SDLs) in two F2 populations of rice (Oryza sativa L.), i.e. Taichung65/Bhadua (TB; japonica-indica cross) and CPSLO17/W207-2 (CW; japonica-japonica). Of the four parents, W207-2 and Bhadua were found to be spikelet semi-sterile and stably inherited through selfing, and spikelet fertility segregated in the two populations. Therefore, recombination frequencies were recalculated after mapping the SDLs by using the multipoint method, and the molecular linkage maps of the two F2 populations were constructed to detect QTLs underlying spikelet fertility. As a result, five SDLs in the TB population were mapped on chromosomes 1, 3, 8 and 9, respectively. Two major QTLs underlying spikelet fertility, namely qSS-6a and qSS-8a, were detected on chromosomes 6 and 8, respectively. In the CW population, a total of 12 SDLs were detected on all 12 chromosomes except 1, 5, 7 and 11. Three QTLs underlying spikelet sterility, namely qSS-2, qSS-6b and qSS-8b on chromosomes 2, 6 and 8, were determined on the whole genome scale. Interestingly, both qSS-6a and qSS-6b, detected in the two F2 populations respectively, were located on a similar position as the S5 gene on chromosome 6; while qSS-8a and qSS-8b were also simultaneously detected on similar positions of the short arm of chromosome 8 in the two populations, which should be a new sterility gene showing the same type of zygotic selection. PMID- 16356284 TI - A QTL analysis of female variation contributing to refractoriness and sperm competition in Drosophila melanogaster. AB - Sperm competition is an important fitness component in many animal groups. Drosophila melanogaster males exhibit substantial genetic variation for sperm competitive ability and females show considerable genetic variation for first versus second male sperm use. Currently, the forces responsible for maintaining genetic variation in sperm competition related phenotypes are receiving much attention. While several candidate genes contributing to the variation seen in male competitive ability are known, genes involved in female sperm use remain largely undiscovered. Without knowledge of the underlying genes, it will be difficult to distinguish between different models of sexual selection such as cryptic female choice and sexual conflict. We used quantitative trait locus (QTL) mapping to identify regions of the genome contributing to female propensity to use first or second male sperm, female refractoriness to re-mating, and early life fertility. The most well supported markers influencing the phenotypes include 33F/34A (P2), 57B (refractoriness) and 23F/24A (fertility). Between 10% and 15% of the phenotypic variance observed in these recombinant inbred lines was explained by these individual QTLs. More detailed investigation of the regions detected in this experiment may lead to the identification of genes responsible for the QTLs identified here. PMID- 16356285 TI - Many QTLs with minor additive effects are associated with a large difference in growth between two selection lines in chickens. AB - Two growth-selected lines in chickens have been developed from a single founder population by divergent selection for body weight at 56 days of age. After more than 40 generations of selection they show a nine-fold difference in body weight at selection age and large differences in growth rate, appetite, fat deposition and metabolic characteristics. We have generated a large intercross between these lines comprising more than 800 F2 birds. QTL mapping revealed 13 loci affecting growth. The most striking observation was that the allele in the high weight line in all cases was associated with enhanced growth, but each locus explained only a small proportion of the phenotypic variance using a standard QTL model (1.3 3.1%). This result is in sharp contrast to our previous study where we reported that the two-fold difference in adult body size between the red junglefowl and White Leghorn domestic chickens is explained by a small number of QTLs with large additive effects. Furthermore, no QTLs for anorexia or antibody response were detected despite large differences for these traits between the founder lines. The result is an excellent example where a large phenotypic difference between populations occurs in the apparent absence of any single locus with large phenotypic effects. The study underscores the need for powerful experimental designs in genetic studies of multifactorial traits. No QTL at all would have reached genome-wide significance using a less powerful design (e.g. approx. 200 F2 individuals) regardless of the nine-fold phenotypic difference between the founder lines for the selected trait. PMID- 16356286 TI - A large-scale screening of the normalized mammalian mitochondrial gene expression profiles. AB - Mammalian mitochondrial genomes are organized in a conserved and extremely compact manner, encoding molecules that play a vital role in oxidative phosphorylation (OXPHOS) and carry out a number of other important biological functions. A large-scale screening of the normalized mitochondrial gene expression profiles generated from publicly available mammalian serial analysis of gene expression (SAGE) datasets (over 17.7 millions of tags) was performed in this study. Acquired SAGE libraries represent an extensive range of human, mouse, rat, bovine and swine cell and tissue samples (normal and pathological) in a variety of conditions. Using a straightforward in silico algorithm, variations in total mitochondrial gene expression, as well as in the expression of individual genes encoded by mitochondrial genomes are addressed, and common patterns in the species- and tissue-specific mitochondrial gene expression profiles are discussed. PMID- 16356287 TI - A simulation study on detecting purging of inbreeding depression in captive populations. AB - Inbreeding depression threatens the survival of small populations of both captive and wild outbreeding species. In order to fully understand this threat, it is necessary to investigate what role purging plays in reducing inbreeding depression. Ballou (1997) undertook such an investigation on 25 mammalian populations, using an ancestral inbreeding regression model to detect purging. He concluded that there was a small but highly significant trend of purging on neonatal survival across the populations. We tested the performance of the regression model that Ballou used to detect purging on independently simulated data. We found that the model has low statistical power when inbreeding depression is caused by the build-up of mildly deleterious alleles. It is therefore possible that Ballou's study may have underestimated the effects of ancestral inbreeding on the purging of inbreeding depression in captive populations if their inbreeding depression was caused mainly by mildly deleterious mutations. We also developed an alternative regression model to Ballou's, which showed an improvement in the detection of purging of mildly deleterious alleles but performed less well if deleterious alleles were of a large effect. PMID- 16356291 TI - Population promotion of informal self-help strategies for early intervention against depression and anxiety. AB - Much of the burden of depression and anxiety in the population is attributable to subclinical symptoms. Broadening formal health-care systems to treat subclinical depression and anxiety is not the answer to reducing this burden, because health care systems lack the resources even to provide optimal care to clinical cases. The solution proposed is the population-wide dissemination of informal self-help strategies that have evidence for effectiveness. These are highly acceptable to the public, easily applied, inexpensive and may avert the development of many clinical cases. PMID- 16356292 TI - Case management to improve major depression in primary health care: a systematic review. AB - BACKGROUND: Deficits in the care of depression lead to poor medication adherence, which increases the risk of an unfavourable outcome for this care. This review evaluates effects on symptoms and medication adherence of case management in primary health care. METHOD: A systematic literature search was performed. The quality of the studies was rated according to the Cochrane Effective Practice and Organization of Care Group (EPOC) criteria. To conduct a subgroup analysis interventions were classified as either 'standard' or 'complex' case management. RESULTS: Thirteen studies met the inclusion criteria. In a meta-analysis we calculated a standard mean difference/effect size on symptom severity after 6-12 months of -0.40 (95% CI -0.60 to -0.20). Patients in the intervention groups were more likely to achieve remission after 6-12 months [relative risk (RR) 1.39, 95% CI 1.30-1.48]. The relative risk for clinical response was 1.82 (95% CI 1.68 2.05). Patients in intervention groups had better medication adherence than the control group (RR 1.5, 95% CI 1.28-1.86). We found heterogeneous results when assessing effects of different types of intervention. CONCLUSIONS: We conclude that case management improves management of major depression in primary health care settings. PMID- 16356293 TI - Short-term effects of enhanced treatment for depression in primary care: results from a randomized controlled trial. AB - BACKGROUND: Depression is a highly prevalent, often recurring or persistent disorder. The majority of patients are initially seen and treated in primary care. Effective treatments are available, but possibilities for providing adequate follow-up care are often limited in this setting. This study assesses the effectiveness of primary-care-based enhanced treatment modalities on short term patient outcomes. METHOD: In a randomized controlled trial we evaluated a psycho-educational self-management intervention. We included 267 adult patients meeting criteria for a DSM-IV diagnosis of major depressive disorder, assessed by a structured psychiatric interview. Patients were randomly assigned to: the Depression Recurrence Prevention (DRP) program (n=112); a combination of the DRP program with psychiatric consultation (PC+DRP, n=39); a combination with brief cognitive behavior therapy (CBT+DRP, n=44); and care as usual (CAU, n=72). Follow up assessments were made at 3 months (response 90%) and 6 months (85%). RESULTS: Patient acceptance of enhanced care was good. The mean duration of the index episode was 11 weeks (S.D.=9.78) and similar in CAU and enhanced care. Recovery rate after 6 months was 67% overall; 17% of all participants remained depressed for the entire 6-month period. CONCLUSION: Enhanced care did not result in better short-term outcomes. We found no evidence that the DRP program was more effective than CAU and no indications for added beneficial effects of either the psychiatric evaluation or the CBT treatment to the basic format of the DRP program. Observed depression treatment rates in CAU were high. PMID- 16356294 TI - Gender differences in the prevalence of DSM-IV and ICD-10 PTSD. AB - BACKGROUND: Gender differences in the prevalence of post-traumatic stress disorder were examined by analysing discrepancies between the DSM-IV and ICD-10 diagnostic systems. METHOD: Data from the Australian National Survey of Mental Health and Well-Being (n=10641) were analysed at the diagnostic, criterion and symptom level for DSM-IV and ICD-10 PTSD for males versus females. RESULTS: While there was a significant gender difference in the prevalence of PTSD for ICD-10, no such difference was found for DSM-IV. The pattern of gender difference at the diagnostic level was mirrored in the pattern of gender differences at the criterion level for both DSM-IV and ICD-10. Females only endorsed three symptoms at a significantly higher rate than males. For all other symptoms, endorsement was equal. This apparently small gender difference at the symptom level was sufficient to cause the gender difference at the diagnostic level for ICD-10, but not DSM-IV because of the different manner in which symptoms are configured into criteria in each of the diagnostic systems. CONCLUSIONS: Gender differences in ICD-10 PTSD but not in DSM PTSD diagnoses are attributable in this study to different patterns of endorsement of symptoms by males and females. Possible reasons for the differential endorsement of symptoms and implications for the use of epidemiological instruments are discussed. PMID- 16356295 TI - The impact of cognitions on the development of panic and somatoform disorders: a prospective study in patients with vestibular neuritis. AB - BACKGROUND: There is a high incidence of panic and somatoform disorders after vestibular neuritis. However, the occurrence of psychiatric disorders has been shown to be unrelated to persistent functional vestibular impairment. The aim of the present study was to examine the role played by cognitions in the development of panic and somatoform disorders. METHOD: In a 2-year prospective study, cognitions were recorded at various stages using the Agoraphobic Cognitions Questionnaire and Body Sensations Questionnaire. Our analysis focused on whether body-related anxiety or panic-related thoughts can predict the development of a panic or somatoform disorder. RESULTS: Fear arising on the first day of an acute vestibular episode did not predict the development of panic or somatoform disorders. One week after the dysfunction, however, the fear of vertigo was a significant predictor, explaining 20% of the variance in the development of either disorder. After 6 weeks, persistent fear of vertigo or vomiting predicted approximately 30% of the variance, and after 6 months panic-related thoughts predicted 40% of the variance and, with the inclusion of body-related fears, as much as 60% of the variance in the development of panic or somatoform disorders. CONCLUSION: Our data confirm prospectively a number of fundamental assumptions of cognitive theory concerning the development of anxiety disorders. Subjects who experience vertigo as particularly alarming focus more intensely than other patients on the negative symptoms they perceive as being related to the disorder. Patients with panic-related cognitions were prone to develop panic or somatoform disorders. PMID- 16356296 TI - Analgesic effect of electroacupuncture on complete Freund's adjuvant-induced inflammatory pain in mice: a model of antipain treatment by acupuncture in mice. AB - Electroacupuncture (EA) was applied bilaterally to the acupoints of Zu-san-li (ST 36) and Kun-lun (BL-60) in the hindlimbs of mice. The therapeutic effect of EA on inflammatory pain induced by an ipsilateral injection of complete Freund's adjuvant (CFA) into the right paw of the mouse was investigated in this study. The time of paw-withdrawal latency (PWL) was used as an indicator for judging the intensity of the pain induced by the CFA injection. The EA effects were divided into immediate (PWL tests within 2 h after EA treatment) and cumulative (PWL tests during and after repetitive EA treatments for 3 weeks) effects. As immediate effects, PWL was significantly shortened in the CFA-injected paw, but was again prolonged 20 min after an EA treatment and lasted until 30 min after. As cumulative effects, PWL was significantly shortened in the CFA-injected paw, but recovered from the 2nd to the 8th day during repetitive EA treatments. No such effects could be observed after sham EA treatment, which resulted in behavior similar to that in untreated animals. These results demonstrate that the CFA-induced inflammatory pain in mice is an ideal model system for the investigation of EA effects and may serve as a valuable reference for the clinical treatment of inflammatory pain in human beings. Furthermore, the mouse pain model opens the possibility to apply the investigation also to transgenic mice. PMID- 16356297 TI - Noninvasive visualization of apoptosis using radiolabeled annexin v could predict response to chemotherapy. PMID- 16356298 TI - Lessons learned from Mother Nature. PMID- 16356300 TI - Capecitabine/Oxaliplatin combinations in advanced colorectal cancer: summary of recent randomized studies. PMID- 16356301 TI - Current data with HMG-CoA reductase inhibitors (statins) for colorectal cancer prevention. PMID- 16356302 TI - Phase I/II dose-escalation study of pemetrexed plus irinotecan in patients with advanced colorectal cancer. AB - BACKGROUND: Pemetrexed and irinotecan have demonstrated antitumor activity as single agents in lung, pancreatic, breast, and colorectal cancer (CRC). The distinct mechanisms of action and patterns of resistance displayed by pemetrexed and irinotecan make them attractive agents for combination therapy. PATIENTS AND METHODS: This phase I/II, nonrandomized, open-labeled, single-arm study was composed of 3 segments. The initial phase II portion of the study enrolled 23 patients with advanced CRC who had received 1 previous dose of 5-fluorouracil (5 FU)-based chemotherapy for advanced disease. Because of poorer than anticipated efficacy, a phase I dose-escalation study using vitamin supplementation (n = 12) was added to the original protocol. The phase II dose-escalation portion of the study enrolled 36 patients (64% with previous oxaliplatin-based therapy) who received pemetrexed 500 mg/m2 followed by irinotecan 300 mg/m2 on day 1, every 21 days. RESULTS: For the 35 evaluable patients in the phase II dose-escalation study, the objective response rate was 11.4% (95% confidence interval, 3.2% 26.7%); there was 1 patient with a complete response, 3 with partial responses, and 17 with stable disease. Three of four responders had received previous oxaliplatin-based combination therapy. Grade 3/4 hematologic toxicities included leukopenia (5.6%), anemia (2.8%), and thrombocytopenia (2.8%). Grade 3/4 nonhematologic toxicities included diarrhea (11.1%), increased aminotransferase levels (8.3%), nausea (8.3%), febrile neutropenia (5.6%), vomiting (5.6%), and reduced creatinine clearance (2.8%). CONCLUSION: Pemetrexed plus irinotecan appears to be at least as active as FOLFIRI (leucovorin/5-FU/irinotecan) for second-line therapy of CRC following 5-FU-based combination chemotherapy. Further studies are warranted. PMID- 16356303 TI - FOLFOX alternated with FOLFIRI as first-line chemotherapy for metastatic colorectal cancer. AB - BACKGROUND: 5-fluorouracil (5-FU), irinotecan, and oxaliplatin are the most active drugs in advanced colorectal cancer (CRC), and survival is improved with patient exposure to all of them. The efficacy and safety of an alternating schedule of continuous-infusion 5-FU with leucovorin (LV) plus oxaliplatin (ie, FOLFOX regimen) or irinotecan (ie, FOLFIRI regimen) was assessed in the first line setting. PATIENTS AND METHODS: Seventy-nine patients with previously untreated, unresectable CRC were included. Treatment consisted of 5-FU/LV (de Gramont schedule) plus oxaliplatin (85 mg/m2) alternated biweekly with the same 5 FU/LV regimen plus irinotecan (180 mg/m2). Treatment was maintained until tumor progression or unacceptable toxicity was noted. RESULTS: Median age was 62 years. Performance status was 0/1 in 91% of patients, 63% had 1 organ involved, and 80% had liver metastases. A median of 6 courses per patient (range, 1-9) and a total of 952 infusions were given. The most frequent grade 3/4 toxic events were neutropenia (32%), diarrhea (26%), and asthenia (7%). Grade 1/2 neurotoxicity was seen in 59% of cases, but no grade 3/4 neurotoxicity was observed. There were no toxic deaths. An objective response rate of 54% (4 complete responses plus 39 partial responses) was attained. Median time to progression and overall survival were 13 months and 18 months, respectively. CONCLUSION: This alternating schedule is active, with efficacy results similar to those seen with sequential protocols, the advantages of less toxicity, and 100% patient exposure to irinotecan and oxaliplatin. PMID- 16356304 TI - Contemporary applications of transanal endoscopic microsurgery: technical innovations and limitations. AB - PURPOSE: Transanal endoscopic microsurgery (TEM) is a minimally invasive procedure used to transanally excise select benign and malignant tumors of the rectum. In properly selected patients, TEM can provide for decreased postoperative morbidity without compromising oncologic outcome. This report summarizes the recent literature concerning TEM, comprehensively analyzes the authors' experience with TEM, and describes recent technical innovations and indications. PATIENTS AND METHODS: Thirty-two consecutive patients scheduled for TEM were identified from our prospectively maintained colorectal service database. Clinicopathologic factors, postoperative complications, and oncologic outcomes were analyzed for all patients. In addition, a PubMed literature search was performed with use of the key words "transanal endoscopic microsurgery," "TEM," "rectal tumor," and "rectal cancer." RESULTS: Transanal endoscopic microsurgery was performed for rectal adenocarcinoma (n = 17; 53%), adenoma (n = 12; 38%), and carcinoid tumors (n = 3; 9%). Median tumor location was 9 cm from the anal verge (range, 3-15 cm). Of the 32 attempted TEM procedures, 27 (84%) were completed. Reasons for inability to complete TEM included narrow rectal lumen or contour of bony pelvis prohibiting passage of the operating proctoscope into the upper rectum and inability to maintain the proctoscope in the rectal lumen with carbon dioxide insufflation because of the distal location of the tumor. Innovations used in the excision of rectal tumors via TEM included the use of the harmonic scalpel, closure of the rectal defect with an extracorporeal slip knot, and a hybrid approach incorporating TEM and traditional transanal techniques. CONCLUSION: Transanal endoscopic microsurgery provides for low morbidity and does not appear to impair oncologic outcome in properly selected patients. PMID- 16356305 TI - Clinicopathologic significance of synchronous and metachronous adenomas in colorectal cancer. AB - PURPOSE: Colorectal cancers (CRCs) evolve from a multiple-step tumorigenesis and, morphologically, are characterized by adenoma. Colorectal cancers with adenomas have distinct clinical features, including reports of improved survival. It is hypothesized that this survival advantage is related to biologic differences in CRC with adenomas rather than earlier diagnosis or earlier stage of disease presentation. PATIENTS AND METHODS: A retrospective chart review of 569 patients treated from 1983 through 2002 was conducted. Data on age, sex, and survival; CRC stage, location, and recurrence; adenoma number, size, histology, and location; and colonoscopy history were analyzed. RESULTS: The mean patient age was 62 years (range, 17-90 years), and 54% of patients were men. The majority of CRCs were left-sided (67%). The American Joint Committee on Cancer stage distribution was 0/I (12%), II (21%), III (34%), and IV (33%). Colorectal cancer with synchronous adenoma was seen in 33% of cases; overall, CRC with adenoma comprised 42% of cases. The event-free survival and overall survival favored CRC with adenoma. After adjusting for age, disease stage, sex, and total number of colonoscopic examinations, the relative risk for an event was 1.51 (P < 0.003) for patients without adenomas versus those with adenomas. CONCLUSION: Colorectal cancer with adenoma represents a distinct population of patients with CRC. The apparent association seems to confer a survival advantage that is not based on age, sex, or disease stage. The survival benefit, although slightly less dramatic, remained significant even when controlled for the number of colonoscopies. PMID- 16356306 TI - Outcomes in elderly patients with advanced colorectal cancer treated with capecitabine: a population-based analysis. AB - BACKGROUND: Toxicity concerns impact the delivery of palliative chemotherapy to elderly patients with advanced colorectal cancer (CRC). Capecitabine was approved for funding in the province of British Columbia in the spring of 2002 as an oral chemotherapeutic option for metastatic CRC. PATIENTS AND METHODS: We conducted a population-based study to assess temporal trends in the use of systemic agents in elderly patients. Patients aged > or = 70 years with metastatic CRC diagnosed between January 1, 2000, and December 31, 2000, and between June 1, 2002, and May 31, 2003, were identified through the British Columbia Cancer Agency Registry. The time cohorts were before and after the provincial approval of capecitabine. Data were obtained regarding demographics, systemic therapies, and outcomes. RESULTS: In cohort A, 35 of 89 patients (39%) were treated with chemotherapy. In the treated versus untreated groups of cohort A, 66% and 57% of patients were male, median ages were 73 years and 76 years, and liver metastasis was seen in 69% and 70% of patients, respectively. In cohort B, 36 of 78 patients (46%) were treated with systemic therapy. In the treated versus untreated groups of cohort B, 58% and 40% of patients were male, median ages were 75 years and 78.5 years, and liver metastasis was seen in 78% and 64% of patients. The most common first line chemotherapy regimens used in cohort A included single-agent 5-FU in 66%, irinotecan-based regimens in 17%, and other regimens in 11%. First-line chemotherapy in cohort B included capecitabine in 47%, oxaliplatin-based regimens in 19%, and irinotecan-based regimens in 17%. The median times to treatment failure resulting from toxicity, disease progression, or death were 37 days in cohort A and 61 days in cohort B. Overall survival between the 2 time cohorts did not differ significantly. Toxicities resulting in dose delay and/or reduction were comparable. CONCLUSION: We conclude that, in patients > or = 70 years of age with advanced CRC, single-agent 5-FU and capecitabine were the favored palliative regimens in British Columbia in 2000 and 2002, respectively. Capecitabine was well tolerated, and both treatments demonstrated similar survival. There was a trend observed toward a greater proportion of patients being offered systemic therapy in the 2002 cohort; however, the difference was not statistically significant. PMID- 16356307 TI - Oxaliplatin induces a delayed immune-mediated hemolytic anemia: a case report and review of the literature. AB - We report a case of a 59-year-old woman with metastatic carcinoma of the ileocecal region who received FOLFOX(oxaliplatin/leucovorin/5-fluorouracil) and bevacizumab therapy and exhibited a partial remission with minimal side effects. She developed a mild self-limited episode of immune-mediated hemolytic anemia during her 16th cycle of chemotherapy, which precluded her from receiving further oxaliplatin. We review the literature on oxaliplatin-induced immune-mediated hemolysis, including its mechanism, presenting symptoms, laboratory features, management, and implications for future therapy. PMID- 16356308 TI - Thyroid metastasis from colorectal cancer: role of [18F]-fluoro-2-deoxy-D-glucose positron emission tomography. AB - We present a patient with colon carcinoma metastatic to the thyroid. Review of the literature reveals only a few reports of metastatic colorectal carcinoma to the thyroid. Metastatic tumors of the thyroid are no longer considered rare. Unfortunately, they often remain undetected because only a small minority of patients present with a mass lesion or enlargement of the gland. This is further evidenced by the fact that most reports come from autopsy series. Establishing this diagnosis is important because metastatic deposits in the thyroid can sometimes cause respiratory compromise as well as thyrotoxicosis. PMID- 16356309 TI - Cancer and Leukemia Group B/Southwest Oncology Group trial 80405: a phase III trial of chemotherapy and biologics for patients with untreated advanced colorectal adenocarcinoma. PMID- 16356310 TI - The need for cost-effectiveness studies in telemedicine. AB - Telemedicine has the potential substantially to improve the delivery of health care. However, cost-effectiveness studies are needed to help define the appropriate scope and application of telemedicine in different settings. Reports on the evaluation of telemedicine are dominated by technical and feasibility studies. Such studies may be very helpful for initial decision making. However, any cost information at this level tends to be very preliminary and often concerned with making a case to proceed further. Decision makers will wish for further information as the telemedicine application is introduced, to consider its effectiveness - its performance under routine conditions. Without information on the costs and effectiveness of telemedicine services, decision makers run the risk of supporting telemedicine systems that are not responsive to health-care needs or which do not provide cost-effective services. The most immediate needs seem to be improvements in the conduct and reporting of studies, and additional information on the performance of telemedicine in routine practice. Investigators need to provide transparent accounts of their studies, describing in detail the approaches taken, sources of data and assumptions made, and indicating the reliability of their results. Decisions may have to be made on the basis of limited studies, but sufficient detail must be made available to decision makers. PMID- 16356311 TI - Telemedicine in the future. AB - Telemedicine can provide a compelling alternative to conventional acute, chronic and preventive care, and can improve clinical outcomes. In the industrialized world, it is likely that telemedicine will continue to move healthcare delivery from the hospital or clinic into the home. In the developing world or in regions with limited infrastructure, telemedicine will mainly be used in applications that link providers based at health centres, referral hospitals and tertiary centres. The future of telemedicine will depend on: (1) human factors, (2) economics and (3) technology. Behaviours related to technology affect change at the individual, organizational and societal level. Personnel shortages and decreasing third-party reimbursement are significant drivers of technology enabled health care in the industrialized world, particularly in the areas of home care and self-care. We can safely assume that developments in mobile communications, sensor devices and nanotechnology will alter the way that health care is delivered in the future. The growth and integration of information and communication technologies into health-care delivery holds great potential for patients, providers and payers in health systems of the future. Perhaps the most difficult question to answer, however, is 'When will telemedicine become part of the standard of care?' PMID- 16356312 TI - Telemedicine for eye care. AB - Tele-ophthalmology has been employed mainly for patients in under-served rural areas in need of specialty care, but other applications such as telementoring have also been used. In certain populations, cost containment is a significant issue and telemedicine is a solution. Tele-ophthalmology can be performed in realtime, by store-and-forward mode, or by hybrid techniques. After appropriate modification, a range of peripherals may be used for tele-ophthalmology, including the direct ophthalmoscope, indirect ophthalmoscope, slit lamp or retinal camera. Tele-ophthalmology applications include: detecting, screening and diagnosing diabetic retinopathy; anterior segment imaging; glaucoma screening; low vision consultation; telementoring. Tele-ophthalmology shows great promise for improving patient care and increasing access to specialty care not available in under-served areas. In developing countries tele-ophthalmology may be a cost effective method by which richer countries can assist them. PMID- 16356313 TI - The Arizona Telemedicine Program business model. AB - The Arizona Telemedicine Program (ATP) was established in 1996 when state funding was provided to implement eight telemedicine sites. Since then the ATP has expanded to connect 55 health-care organizations through a membership programme formalized through legal contracts. The ATP's membership model is based on an application service provider (ASP) concept, whereby organizations can share services at lower cost; that is, the ATP acts as a broker for services. The membership fee schedule is flexible, allowing clients to purchase only those services desired. An annual membership fee is paid by every user, based on the services requested. The membership programme income has provided a steady revenue stream for the ATP. The membership-derived revenue represented 30% of the ATP's 2.6 million dollars total income during fiscal year 2003/04. PMID- 16356314 TI - Bases for assessments made by telephone advice nurses. AB - Telephone advice nursing includes triage, advice, referral, information and coordination. The aim of the study was to explore what telephone nurses base their assessments on. We conducted 14 interviews with seven telephone nurses at a health-care call centre in Sweden. Two authentic calls per nurse were used in stimulated recall interviews, where the nurses commented on the basis for their assessments. A qualitative manifest content analysis was employed. Three major categories emerged in the analysis: care-seeker, e.g. 'symptomatic sounds'; nurse, e.g. 'nurse's own experience'; and organization, e.g. 'health-care accessibility'. The findings show that the telephone advice nurses' bases for assessments appear to be very broad. They include both verbally and nonverbally communicated information, and care-seeker-, nurse- and certain organization related factors influence the assessments. We found that an individualistic view of the care-seeker seems to dominate the assessments in non-urgent calls to a health-care call centre. PMID- 16356315 TI - An evaluation of a surgical telepresence system for an intrahospital local area network. AB - We evaluated a digital telepresence system in an operating theatre (OR) environment which enabled a consultant to join the surgical team from a remote site by audiovisual communication. The system is based on video transmission using a streaming technique, with a server and a client connected via a local area network (LAN). Two cameras can be remotely controlled: one camera is built into the OR lamp and a second, laparoscopic camera is mounted on a robotic arm. Another feature of the system is teledemonstration, which permits the remote consultant to demonstrate points of particular interest. We evaluated the system clinically in 237 cases. In 28 cases (12%), telepresence could not be established for various reasons, mainly human failure. In 42 cases (18%), the full potential of telepresence was used. Technical evaluation showed that a data rate of 2 Mbit/s provides sufficient audio and video quality, as well as reliable teledemonstration. The data transmission delay was acceptable for clinical purposes (video 0.92 s, audio 0.6 s from OR to client, audio 0.7 s from client to OR). The study showed that telepresence is a promising means of providing highly specialized expertise within the OR. PMID- 16356316 TI - User satisfaction with a telemedicine amputee clinic in Saskatchewan. AB - A group of 15 patients with amputee-related diagnoses were given a satisfaction survey after telemedicine assessment. Most of the videoconferencing sessions used an IP connection at 768 kbit/s. The patients were seen at four sites. The average connection time was less than 5 min and the average time for a session was approximately 40 min. Thirteen questions required scaled responses (poor, fair, good, excellent) and two required yes/no answers. The 13 categories broadly related to satisfaction with the telemedicine service and the quality of specialist care. In all categories, 97% of the responses fell in the good to excellent range. Concerns were raised about ease of access to local telemedicine sites, connection waiting times and lack of familiarity with telemedicine technology. The study showed that telemedicine was acceptable to patients with amputations and provided a reliable assessment of the amputee. PMID- 16356317 TI - Delivery of rural and remote health care via a broadband Internet Protocol network - views of potential users. AB - We asked the views of potential users of a proposed Canadian broadband Internet Protocol (IP) network for health, the Alberta SuperNet. The three user groups were drawn from the public, provider and private sectors. In all, 35 health sector participants were selected (17 government, nine health-care organizations, five providers/practitioners and four private sector). The questionnaire was Web based, semistructured and self-administered. It consisted of four major areas: value, readiness, effect on usual care and limitations. A total of 28 (80%) individuals responded to the questionnaire: 21 (81%) were from the public sector (three provincial, nine regional and nine organizational), three (60%) were from the provider sector and four (100%) were from the private sector. Overall, the items related to health services and health human resources were considered to be the most valuable to rural communities. Respondents identified the expansion of telehealth services as the most important, except those from the private sector, who ranked this a close second. The health system's move to the use of electronic health records was ranked second in importance by all respondents. The private sector respondents viewed all user groups to be generally less ready (mean score 2.5 on a seven-point scale from 1 = not ready to 7 = ready), while the public sector respondents were the most optimistic (mean score 4.0). Specific socioeconomic impact data were limited. The top-ranked disadvantage of the 10 suggested was that 'Changes in health-service delivery practices and/or processes will be required'. PMID- 16356321 TI - Graded visual attention modulates brain responses evoked by task-irrelevant auditory pitch changes. AB - Previous studies suggested that auditory change-specific neural responses are attention-independent and reflect central auditory processing. The automaticity of the brain's response to infrequent changes in pitch within a series of auditory tone pips was examined in parallel functional magnetic resonance imaging (fMRI) and event-related potential (ERP) studies. Subjects performed a continuous perceptual-motor visual tracking task at two levels of difficulty while simultaneously hearing a series of task-irrelevant standard tone pips and infrequent pitch-deviant tones. fMRI results revealed that the unattended pitch deviant tones strongly activated superior temporal and frontal cortical regions. These activations were significantly modulated by the tracking difficulty of the primary task. ERP results revealed that the amplitude of the scalp-negative component evoked by deviant tones (MMN) was attenuated during the more difficult tracking task. Our results demonstrate that the brain's response to task irrelevant sensory changes is strongly influenced by intermodal attentional demands. PMID- 16356322 TI - Different effects of exogenous cues in a visual detection and discrimination task: delayed attention withdrawal and/or speeded motor inhibition? AB - Several studies examining spatial attention have found a discrepancy regarding the effects of exogenous cues on reaction times in visual detection and discrimination tasks. Namely, across a wide range of cue-target intervals, responses are slower for targets at cued than at uncued locations (inhibition of return) in detection tasks, whereas responses are faster for targets at cued than at uncued locations (facilitation) in discrimination tasks. Two hypotheses were proposed to account for this discrepancy. First, attention may dwell much longer on the exogenously cued location in discrimination tasks because stimuli have to be identified (i.e., the delayed attention withdrawal hypothesis). Secondly, due to increased motor preparation in detection tasks, cue-induced motor inhibition may rise much faster in these tasks than in discrimination tasks (i.e., the speeded motor inhibition hypothesis). We examined to what extent these hypotheses can account for effects of exogenous cues in a detection and discrimination task on the extrastriate P1 component, and the onset of motor activation, as indexed by the lateralized readiness potential. Some support was found for the delayed attention withdrawal hypothesis, as task-dependent cueing effects were found on the P1 component. Other aspects of our data, however, indicate that motor inhibition is also involved. Based on these findings, we propose that effects of exogenous cues in detection and discrimination tasks are determined by the interplay between two mechanisms, of which the time courses of activation may be modulated by the specific setting. PMID- 16356323 TI - Orthographic distinctiveness and semantic elaboration provide separate contributions to memory. AB - Orthographic distinctiveness and semantic elaboration both enhance memory. The present behavioral and functional magnetic resonance imaging (fMRI) studies examined the relationship between the influences of orthographic distinctiveness and semantic elaboration on memory, and explored whether they make independent contributions. As is typical for manipulations of processing levels, words studied during semantic encoding were better remembered than words studied during nonsemantic encoding. Notably, orthographically distinct words were better recalled and received more remember responses on recognition memory tests than orthographically common words regardless of encoding task, suggesting that orthographic distinctiveness has an additive effect to that of semantic elaboration on memory. In the fMRI study, orthographic distinctiveness and semantic elaboration engaged separate networks of brain regions. Semantic elaboration modulated activity in left inferior prefrontal and lateral temporal regions. In contrast, orthographic distinctiveness modulated activity in distinct bilateral inferior prefrontal, extrastriate, and parietal regions. Orthographic distinctiveness and semantic elaboration appear to have separate behavioral and functional-anatomic contributions to memory. PMID- 16356324 TI - Conceptual representations of action in the lateral temporal cortex. AB - Retrieval of conceptual information from action pictures causes greater activation than from object pictures bilaterally in human motion areas (MT/MST) and nearby temporal regions. By contrast, retrieval of conceptual information from action words causes greater activation in left middle and superior temporal gyri, anterior and dorsal to the MT/MST. We performed two fMRI experiments to replicate and extend these findings regarding action words. In the first experiment, subjects performed conceptual judgments of action and object words under conditions that stressed visual semantic information. Under these conditions, action words again activated posterior temporal regions close to, but not identical with, the MT/MST. In the second experiment, we included conceptual judgments of manipulable object words in addition to judgments of action and animal words. Both action and manipulable object judgments caused greater activity than animal judgments in the posterior middle temporal gyrus. Both of these experiments support the hypothesis that middle temporal gyrus activation is related to accessing conceptual information about motion attributes, rather than alternative accounts on the basis of lexical or grammatical factors. Furthermore, these experiments provide additional support for the notion of a concrete to abstract gradient of motion representations with the lateral occipito-temporal cortex, extending anterior and dorsal from the MT/MST towards the peri-sylvian cortex. PMID- 16356325 TI - Automatic priming of semantically related words reduces activity in the fusiform gyrus. AB - We used rapid, event-related fMRI to identify the neural systems underlying object semantics. During scanning, subjects silently read rapidly presented word pairs (150 msec, SOA = 250 msec) that were either unrelated in meaning (ankle carrot), semantically related (fork-cup), or identical (crow-crow). Activity in the left posterior region of the fusiform gyrus and left inferior frontal cortex was modulated by word-pair relationship. Semantically related pairs yielded less activity than unrelated pairs, but greater activity than identical pairs, mirroring the pattern of behavioral facilitation as measured by word reading times. These findings provide strong support for the involvement of these areas in the automatic processing of object meaning. In addition, words referring to animate objects produced greater activity in the lateral region of the fusiform gyri, right superior temporal sulcus, and medial region of the occipital lobe relative to manmade, manipulable objects, whereas words referring to manmade, manipulable objects produced greater activity in the left ventral premotor, left anterior cingulate, and bilateral parietal cortices relative to animate objects. These findings are consistent with the dissociation between these areas based on sensory- and motor-related object properties, providing further evidence that conceptual object knowledge is housed, in part, in the same neural systems that subserve perception and action. PMID- 16356326 TI - An effect of dopamine depletion on decision-making: the temporal coupling of deliberation and execution. AB - When a decision between alternative actions has to be made, the primate brain is able to uncouple motor execution from mental deliberation, providing time for higher cognitive processes such as remembering and reasoning. The mental deliberation leading to the decision and the motor execution applying the decision are likely to involve different neuronal circuits linking the basal ganglia and the frontal cortex. Behavioral and physiological studies in monkeys indicate that dopamine depletion may result in a loss of functional segregation between these circuits, hence, in interference between the deliberation and execution processes. To test this hypothesis in humans, we analyzed the movements of parkinsonian patients in a go/no-go task, contrasting periods of uncertainty with periods of knowledge about the rule to be applied. Two groups of patients were compared to healthy subjects: one group was treated with dopaminergic medication and the other with deep brain stimulation; both groups were also tested without any treatment. In healthy subjects, the movement time was unaffected by uncertainty. In untreated patients, the movement time increased with uncertainty, reflecting interference between deliberation and execution processes. This interference was fully corrected with dopaminergic medication but was unchanged with deep brain stimulation. Moreover, decision-related hesitations were detectable in the movements of dopamine-depleted patients, revealing a temporal coupling of deliberation and execution. We suggest that such coupling may be related to the loss of dopamine-mediated functional segregation between basal ganglia circuits processing different stages of goal-directed behavior. PMID- 16356328 TI - Pushing around the locus of selection: evidence for the flexible-selection hypothesis. AB - Attention operates at an early stage in some experimental paradigms and at a late stage in others, which suggests that the locus of selection is flexible. The present study was designed to determine whether the locus of selection can vary flexibly within a single experimental paradigm as a function of relatively modest variations in stimulus and task parameters. In the first experiment, a new method for assessing the locus of selection was developed. Specifically, attention can influence perceptual encoding only if it is directed to the target before a perceptual representation of the target has been formed, whereas attention can influence postperceptual processes even if attention is cued after perception is complete. Event-related potentials were used to confirm the validity of this method. The subsequent experiments used cueing tasks in which subjects were required to perceive and remember a set of objects, and the difficulty of the perception and memory components of the task were varied. When the task overloaded perception but not working memory, attention influenced the formation of perceptual representations but not the storage of these representations in memory; when the task overloaded working memory but not perception, attention influenced the transfer of perceptual representations into memory but not the formation of the perceptual representations. Thus, attention operates to select relevant information at whatever stage or stages of processing are overloaded by a particular stimulus-task combination. PMID- 16356327 TI - The cerebral response during subjective choice with and without self-reference. AB - The anterior medial prefrontal (AMPFC) and retrosplenial (RSC) cortices are active during self-referential decision-making tasks such as when participants appraise traits and abilities, or current affect. Other appraisal tasks requiring an evaluative decision or mental representation, such as theory of mind and perspective-taking tasks, also involve these regions. In many instances, these types of decisions involve a subjective opinion or preference, but also a degree of ambiguity in the decision, rather than a strictly veridical response. However, this ambiguity is generally not controlled for in studies that examine self referential decision-making. In this functional magnetic resonance imaging experiment with 17 healthy adults, we examined neural processes associated with subjective decision-making with and without an overt self-referential component. The task required subjective decisions about colors-regarding self-preference (internal subjective decision) or color similarity (external subjective decision) under conditions where there was no objectively correct response. Results indicated greater activation in the AMPFC, RSC, and caudate nucleus during internal subjective decision-making. The findings suggest that self-referential processing, rather than subjective judgments among ambiguous response alternatives, accounted for the AMPFC and RSC response. PMID- 16356329 TI - Effects of divided attention on fMRI correlates of memory encoding. AB - Performing a secondary task concurrently with a study task has a detrimental effect on later memory for studied items. To investigate the mechanisms underlying this effect, the processing resources available for an incidental encoding task were varied by manipulating secondary task difficulty. fMRI data were acquired as volunteers (n = 16) made animacy decisions to visually presented study words while concurrently performing either an easy or a hard auditory monitoring task. "Subsequent memory effects"--greater activity at study for words later remembered versus words later forgotten--were identified in the left ventral inferior frontal gyrus and the left anterior hippocampus. These effects did not vary according to whether the encoding task was performed concurrently with the easy or the hard secondary task. However, as secondary task difficulty increased, study-item activity declined and auditory-item activity increased in dorsolateral prefrontal and superior parietal regions that have been implicated in the support of executive and control functions. The findings suggest that dividing attention during encoding influences the probability of engaging the encoding operations that support later episodic memory, but does not alter the nature of the operations themselves. The findings further suggest that the probability of engaging these encoding operations depends on the level of general processing resources engaged in service of the study task. PMID- 16356330 TI - Attention and sensory gain control: a peripheral visual process? AB - Attention-related sensory gain control in human extrastriate cortex is believed to improve the acuity of visual perception. Yet given wide variance in the spatial resolution of vision across the retina, it remains unclear whether sensory gain operates homogenously between foveal and nonfoveal retinotopic locations. To address this issue, we used event-related potentials (ERPs) in a variant of the canonical spatial attention task. Participants were cued to expect targets at either fixation (foveal targets) or at a location several degrees above fixation (parafoveal targets). At both target locations, manual reaction times were shorter for cued relative to uncued targets, indicating that attention was consistently oriented to the cued location. Nevertheless, attention-related increases in sensory-evoked cortical activity were only observed at the parafoveal target location, as measured by the amplitude of the lateral occipital P1 ERP component. A second experiment replicated this data pattern using targets with lower stimulus contrast, indicating that the absence of a P1 effect for foveal targets could not be attributed to a saturated P1 response under higher contrast stimulus conditions. When considered in light of retinogeniculate projections to cortex showing systematic changes in their physiological organization beginning within a degree of visual angle of the fovea, our findings support the proposal that the strategic functions of visual attention may vary with the retinotopic location involved. PMID- 16356331 TI - Anticipatory gain modulation in preparation for smooth pursuit eye movements. AB - We studied the effect of the probability of required tracking on the gain of visuomotor transmission for pursuit initiation in monkeys. We recorded the ocular responses to a brief movement (perturbation) of a target located at an eccentric position from the central fixation spot. As soon as the central fixation spot went off, the animal was required to make a saccade to the target if it remained stationary or to track if it moved. The probability of an upcoming ramp motion of the target (horizontal, 20 degrees /sec), requiring pursuit, was varied (target motion probability [TMP] = 0, 0.25, 0.5, 0.75, and 1, which was fixed in a block). We found that the magnitude of the response to the perturbation increased gradually as the TMP increased. The initial pursuit response and the perturbation response showed very similar dependence on the TMP, suggesting that the response to the perturbation could be used as an index of the gain of visuomotor transmission for pursuit initiation. We also found that the changes in the ocular responses after the TMP was changed from one probability to another occurred rapidly (decay constant of only a few trials). These results suggest that the gain of visuomotor transmission in preparing for pursuit is rapidly modulated in accordance with the anticipated future need for pursuit. PMID- 16356332 TI - Phase synchronization analysis of EEG during attentional blink. AB - The attentional blink (AB) phenomenon occurs when perceivers must report two targets embedded in a sequence of distracters; if the first target precedes the second by 200-600 msec, the second one is often missed. We investigated AB by measuring dynamic cross-lag phase synchronization for 565 electrode pairs in 40 Hz-range EEG. Phase synchrony, on average, was higher in experimental conditions, where two targets are reported, than in control conditions, where only the second target is reported. The effect occurred in electrode pairs covering the whole head. Timing of the synchrony was crucial: Brief episodes of enhanced synchrony occurred 100-500 msec before expected target onset in AB conditions where the second target was correctly reported. These results show that intrinsic brain dynamics produce anticipatory synchronization in transient assemblies of cortical areas. Enhanced levels of anticipatory synchronization occur in response to the demands of the task in conditions where the system's limited capacity is under strain. PMID- 16356333 TI - Evaluation of buprenorphine CEDIA assay versus GC-MS and ELISA using urine samples from patients in substitution treatment. AB - As buprenorphine becomes more clinically used in heroin substitution treatment, there is an increasing need for methods suitable for high-volume screening. In this study, a new immunochemical test based on CEDIA technology was evaluated for the use in clinical urine drug testing. The method was compared with an existing ELISA method and a gas chromatography-mass spectrometry (GC-MS) method on urine specimens from patients in heroin substitution treatment. The precision of the CEDIA assay was < 9% both within- and between-day at levels at and above the cutoff limit of 5 microg/L. The concordance in qualitative results with an existing ELISA method was 96.8%. The CEDIA measuring range was extended by diluting urine samples 100-fold with saline, and the results agreed well (slope of regression line was 1.09, r(2) = 0.968) with GC-MS. The sensitivity of CEDIA in detecting authentic specimen containing buprenorphine at levels >or= 5 microg/L was 99.5%. Cross-reactivity causing false-positive response was discovered in patients receiving prescribed dihydrocodeine. The urine concentration of total buprenorphine in urine from patients prescribed daily doses between 0.2 and 24 mg ranged from 0.5 to 2900 microg/L. The concentration of the metabolite norbuprenorphine was usually higher, and the median ratio of buprenorphine to norbuprenorphine was 0.23 (95% were below 1). We conclude that the CEDIA assay is suitable for application in high-volume screening of buprenorphine for urine drug testing. PMID- 16356334 TI - Detection of exogenous GHB in blood by gas chromatography-combustion-isotope ratio mass spectrometry: implications in postmortem toxicology. AB - Because GHB (gamma-hydroxybutyrate) is present in both blood and urine of the general population, toxicologists must be able to discriminate between endogenous levels and a concentration resulting from exposure. In this paper, we propose a procedure for the detection of exogenous GHB in blood by gas chromatography combustion-isotope ratio mass spectrometry (GC-C-IRMS). Following liquid-liquid and solid-phase extractions, GHB is derivatized to GHB di-TMS before analysis by GC-C-IRMS. Significant differences in the carbon isotopic ratio (delta(13)C values > 13.5 per thousand) were found between endogenous and synthetic GHB. Indeed, for postmortem blood samples with different GHB concentrations (range: 13.8-86.3 mg/L), we have obtained GHB delta(13)C-values ranging from -20.6 to 24.7 per thousand, whereas delta(13)C-values for the GHB from police seizure were in the range -38.2 to -50.2 per thousand. In contrast to the use of cut-off concentrations for positive postmortem blood GHB concentrations, this method should provide an unambiguous indication of the drug origin. PMID- 16356336 TI - ELISA on a microchip with a photodiode for detection of amphetamine in plasma and urine. AB - A rapid and sensitive assay was developed for the detection of amphetamine in plasma and urine. The method relies on the principle of competitive ELISA (enzyme linked immunosorbent assay). A flow microchip with a total volume of 7 microL was used for the development of a chemiluminescent ELISA technique. Solutions, samples, and the chemiluminescence substrate were injected by a flow system, and a photodiode detector was used to measure the light intensity. The incubation time of the competitor (competition phase) was reduced to 10 min. Calibration curves corresponding to analyte concentrations ranging from 40 to 1,000 microg/L in urine samples and from 6 to 96 microg/L in plasma samples were obtained. The detection limits were in the region of 20 and 6 microg/L in urine and plasma, respectively. The main focus of the work was on speed, reliability, reproducibility, and operational stability of the assay. This method was proven readily adaptable to automation and provided reproducible results. PMID- 16356335 TI - Simultaneous and sensitive analysis of THC, 11-OH-THC, THC-COOH, CBD, and CBN by GC-MS in plasma after oral application of small doses of THC and cannabis extract. AB - Besides the psychoactive Delta(9)-tetrahydrocannabinol (THC), hashish and marijuana as well as cannabis-based medicine extracts contain varying amounts of cannabidiol (CBD) and of the degradation product cannabinol (CBN). The additional determination of these compounds is interesting from forensic and medical points of view because it can be used for further proof of cannabis exposure and because CBD is known to modify the effects of THC. Therefore, a method for the simultaneous quantitative determination of THC, its metabolites 11-hydroxy Delta(9)-tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-Delta(9) tetrahydrocannabinol (THC-COOH), CBD and CBN from plasma was developed. The method was based on automatic solid-phase extraction with C(18) ec columns, derivatization with N,O-bistrimethylsilyltrifluoroacetamide (BSTFA), and gas chromatography-electron impact ionization-mass spectrometry (GC-EI-MS) with deuterated standards. The limits of detection were between 0.15 and 0.29 ng/mL for THC, 11-OH-THC, THC-COOH, and CBD and 1.1 ng/mL for CBN. The method was applied in a prospective pharmacokinetic study after single oral administration of 10 mg THC alone or together with 5.4 mg CBD in cannabis extract. The maximum plasma concentrations after cannabis extract administration ranged between 1.2 and 10.3 ng/mL (mean 4.05 ng/mL) for THC, 1.8 and 12.3 ng/mL (mean 4.9 ng/mL) for 11-OH-THC, 19 and 71 ng/mL (mean 35 ng/mL) for THC-COOH, and 0.2 and 2.6 ng/mL (mean 0.95 ng/mg) for CBD. The peak concentrations (mean values) of THC, 11-OH THC, THC-COOH, and CBD were observed at 56, 82, 115, and 60 min, respectively, after intake. CBN was not detected. Caused by the strong first-pass metabolism, the concentrations of the metabolites were increased during the first hours after drug administration when compared to literature data for smoking. Therefore, the concentration ratio 11-OH-THC/THC was discussed as a criterion for distinguishing oral from inhalative cannabis consumption. PMID- 16356337 TI - Monitoring of occupational exposure to methylene chloride: sampling protocol and stability of urine samples. AB - A sampling protocol for biomonitoring of the volatile solvent methylene chloride (MeCl(2)) by analysis of urine from exposed workers was established. Storage temperature, sample volume in headspace vial (HSV), and time to sealing HSV on determination of MeCl(2) in urine were evaluated. MeCl(2) was analyzed by a solid phase microextraction technique combined with gas chromatography. Volume of urine in HSV has no effect on MeCl(2) analysis. Delays of 30 and 60 min from collection of urine until sealing the HSV caused 14.47 +/- 6.98% and 26.17 +/- 9.57% decreases from baseline concentration, respectively. MeCl(2) concentration in spiked urine samples stored in sealed HSVs decreased on day 2 and then remained stable for 2 weeks. Refrigeration did not improve recovery although it seems to be associated with less variability. MeCl(2) in urine samples of seven exposed workers was in the range of 0.02-0.06 mg/L. Sampling of MeCl(2)-containing urine should include collection of urine in closed plastic bottles, transfer to HSV within 15 min, sealing and clamping of HSV within 15 s, and storage of HSV in refrigeration until analysis, but no longer than 2 weeks. Standard samples should be prepared on the day of test sample collection and handled under the same conditions. PMID- 16356338 TI - Rapid and simple quantitation of methamphetamine by using a homogeneous time resolved fluoroimmunoassay based on fluorescence resonance energy transfer from europium to Cy5. AB - A rapid and simple homogeneous time-resolved fluoroimmunoassay based on fluorescence resonance energy transfer from europium (Eu) to cyanine dye (Cy5) has been developed for the quantitation of methamphetamine. In this assay, Eu chelate was labeled to a conjugate of methamphetamine and bovine serum albumin (MA-BSA), as an energy donor, and Cy5 was labeled to anti-MA as an energy acceptor. The close proximity between the two labels in the immunocomplex permits energy transfer from the excited Eu(3+) donor. Therefore, by measuring the sensitized emission of Cy5 with the time-resolved assay, immunocomplex of MA-BSA and anti-MA can be measured in the homogeneous solution without separation steps within 30 min. By a competitive immunoassay, MA could be assayed in the range 0.1 1,000 ng/mL. The intra-assay variations were 5.4-14.8% at 5 different concentrations. When urine or serum samples were examined, the quenching of Eu fluorescence was observed, but the acceptor-to-donor ratio constantly depended upon the dilution of samples. Twenty urine samples were assayed, and the data showed a good correlation to those obtained by gas chromatography (r = 0.94). The homogeneous assay using Eu-Cy5 energy transfer is time-saving without any washing procedures and is suitable for screening drugs that are commonly abused. PMID- 16356339 TI - Pesticide hair analysis: development of a GC-NCI-MS method to assess chronic exposure to diazinon in rats. AB - The present study aimed to improve the gas chromatography-mass spectrometry (GC MS) method, already developed in our laboratory, for trace analysis of diazinon in hair. Furthermore, it aimed to compare the disposition of the pesticide in the hair of two different animal species, one susceptible to diazinon toxicity and one resistant, under identical experimental conditions. Sprague Dawley rats were systemically exposed to two dose levels (6 mg/kg/day and 3 mg/kg/day) of the pesticide, through their drinking water, for a period of one and a half months. Hair samples from the back of the rats were removed before commencing the experiment and at the end of the dosing period. Diazinon was selectively isolated from pulverized hair, sample or spiked, by stepwise consequent extractions with methanol and ethyl acetate and quantified by GC-negative chemical ionization-MS. It was found that the concentration of diazinon in the hair of exposed animals was dose dependent and was found to be 0.24 +/- 0.01 ng/mg (n = 5) and 0.53 +/- 0.05 ng/mg (n = 5) for the low and high dosage, respectively. The concentration in both dose groups was much higher than the corresponding rabbit hair (rabbits were exposed to the pesticide under similar experimental conditions) as previously reported. Our results strongly point to the possibility of using hair analysis for low-level exposure monitoring of diazinon. PMID- 16356340 TI - Bone surface and whole bone as biomarkers for acute fluoride exposure. AB - This study compares fluoride concentrations ([F]) in surface and whole bone for up to 27 days following an acute oral dose of F. Four groups of rats received single oral F dose (50 mg/kg body weight), and the control group received deionized water (n = 10/group). Groups were euthanized at 1, 3, 9, or 27 days after F administration. Plasma and femurs were collected. F on the femur surface was removed from a circular area (4.52 mm(2)) by immersion in 0.5M HCl for 15 s. The solution was buffered with total ionic strength adjustment buffer and analyzed with an electrode. The subjacent bone was sectioned and ashed at 600 degrees C. Ash and plasma were analyzed for F with the electrode following hexamethyldisiloxane-facilitated diffusion. Data were analyzed by Kruskall-Wallis and Dunn's test and by linear regression (p < 0.05). Peak plasma and bone surface [F] occurred on day 1 (0.26 +/- 0.14 microg/mL and 1801 +/- 888 microg/g, respectively). Bone surface [F] at 3, 9, and 27 were not statistically different from control. A significant increase in whole bone [F] was observed 3 days after F administration and the [F] remained relatively constant thereafter. The mean (+/- SD) surface/whole bone [F] ratios for the control and F groups were 2.45 +/- 0.98, 3.92 +/- 1.32, 1.61 +/- 0.82, 1.73 +/- 0.39, and 1.09 +/- 0.28, respectively. Plasma and bone surface [F]s were positively correlated (r = 0.74). Thus, bone surface was found to be a suitable biomarker for acute, sublethal F exposure 1 day after F administration. Whole bone [F] were significantly increased at 3, 9, and 27 days after F administration. PMID- 16356342 TI - THC can be detected in brain while absent in blood. PMID- 16356341 TI - A fatal intoxication following the ingestion of 5-methoxy-N,N-dimethyltryptamine in an ayahuasca preparation. AB - A case of a 25-year-old white male who was found dead the morning after consuming herbal extracts containing beta-carbolines and hallucinogenic tryptamines is presented. No anatomic cause of death was found at autopsy. Toxicologic analysis of the heart blood identified N,N-dimethyltryptamine (0.02 mg/L), 5-methoxy-N,N dimethyltryptamine (1.88 mg/L), tetrahydroharmine (0.38 mg/L), harmaline (0.07 mg/L), and harmine (0.17 mg/L). All substances were extracted by a single-step n butyl chloride extraction following alkalinization with borate buffer. Detection and quantitation was performed using liquid chromatography-electrospray mass spectrometry. The medical examiner ruled that the cause of death was hallucinogenic amine intoxication, and the manner of death was undetermined. PMID- 16356354 TI - This world is yours. PMID- 16356355 TI - Program evaluation and chronic diseases: methods, approaches, and implications for public health. PMID- 16356356 TI - The common threads in program evaluation. PMID- 16356357 TI - Using the Office of Management and Budget (OMB) clearance process in program planning and evaluation. PMID- 16356358 TI - The behavioral and clinical effects of therapeutic lifestyle change on middle aged adults. AB - INTRODUCTION: Chronic diseases such as cancer, cardiovascular disease, stroke, and diabetes are responsible for most deaths in the United States. Lifestyle factors--poor nutrition, sedentary living, and tobacco use--appear to play a prominent role in the development of many chronic diseases. This study determined the behavioral and clinical impact of a therapeutic lifestyle-modification intervention on a group of community volunteers. METHODS: Participants included 348 volunteers aged 24 to 81 years from the Rockford, Ill, metropolitan area who participated in a randomized clinical trial. The intervention group attended a 40 hour educational course delivered as lectures during a 4-week period. Participants learned the importance of making better lifestyle choices and how to make improvements in nutrition and physical activity. Changes in nutrition, physical activity behavior, and several chronic disease risk factors were assessed at baseline and 6 months. RESULTS: Intervention participants showed significant 6-month improvement in all nutrition and physical activity measures except calories from protein and whole-grain servings and all clinical measures except blood glucose, total cholesterol, triglycerides, and high-sensitivity C reactive protein. Total cholesterol and low-density lipoprotein cholesterol were worse after 6 months in both groups but only significantly worse in the control group. The control group experienced small but significant improvements in systolic and diastolic blood pressure and high-density lipoproteins. Change-score comparisons between the intervention and control groups were significant for all nutrition and physical activity variables except total steps per week and daily sodium intake and were also significant for the clinical measures of weight, body fat, and body mass index. CONCLUSION: This therapeutic lifestyle-modification program can significantly improve nutrition and physical activity behavior and can reduce many of the risk factors associated with common chronic diseases. PMID- 16356360 TI - Identifying best practices for WISEWOMAN programs using a mixed-methods evaluation. AB - INTRODUCTION: Recommendations on best practices typically are drawn from unique settings; these practices are challenging to implement in programs already in operation. We describe an evaluation that identifies best practices in implementing lifestyle interventions in the Center for Disease Control and Prevention's WISEWOMAN program and discuss our lessons learned in using the approach. METHODS: We used a mixed-methods evaluation that integrated quantitative and qualitative inquiry. Five state or tribal WISEWOMAN projects were included in the study. The projects were selected on the basis of availability of quantitative program performance data, which were used to identify two high-performing and one low-performing site within each project. We collected qualitative data through interviews, observation, and focus groups so we could understand the practices and strategies used to select and implement the interventions. Data were analyzed in a multistep process that included summarization, identification of themes and practices of interest, and application of an algorithm. RESULTS: Pilot testing data collection methods allowed for critical revisions. Conducting preliminary interviews allowed for more in-depth interviews while on site. Observing the lifestyle intervention being administered was key to understanding the program. Conducting focus groups with participants helped to validate information from other sources and offered a more complete picture of the program. CONCLUSION: Using a mixed-methods evaluation minimized the weaknesses inherent in each method and improved the completeness and quality of data collected. A mixed-methods evaluation permits triangulation of data and is a promising strategy for identifying best practices. PMID- 16356359 TI - Using concept mapping to develop a logic model for the Prevention Research Centers Program. AB - INTRODUCTION: Concept mapping is a structured conceptualization process that provides a visual representation of relationships among ideas. Concept mapping was used to develop a logic model for the Centers for Disease Control and Prevention's Prevention Research Centers Program, which has a large and diverse group of stakeholders throughout the United States. No published studies have used concept mapping to develop a logic model for a national program. METHODS: Two logic models were constructed using the data from the concept mapping process and program documents: one for the national level and one for the local level. Concept mapping involved three phases: 1) developing questions to generate ideas about the program's purpose and function, 2) gathering input from 145 national stakeholders and 135 local stakeholders and sorting ideas into themes, and 3) using multivariate statistical analyses to generate concept maps. Logic models were refined using feedback received from stakeholders at regional meetings and conferences and from a structured feedback tool. RESULTS: The national concept map consisted of 9 clusters with 88 statements; the local concept map consisted of 11 clusters with 75 statements. Clusters were categorized into three logic model components: inputs, activities, and outcomes. Based on feedback, two draft logic models were combined and finalized into one for the Prevention Research Centers Program. CONCLUSION: Concept mapping provides a valuable data source, establishes a common view of a program, and identifies inputs, activities, and outcomes in a logic model. Our concept mapping process resulted in a logic model that is meaningful for stakeholders, incorporates input from the program's partners, and establishes important program expectations. Our methods may be beneficial for other programs that are developing logic models for evaluation planning. PMID- 16356361 TI - Using focus groups in the consumer research phase of a social marketing program to promote moderate-intensity physical activity and walking trail use in Sumter County, South Carolina. AB - INTRODUCTION: The use of social marketing approaches in public health practice is increasing. Using marketing concepts such as the "four Ps" (product, price, place, and promotion), social marketing borrows from the principles of commercial marketing but promotes beneficial health behaviors. Consumer research is used to segment the population and develop a strategy based on those marketing concepts. In a community-based participatory research study, 17 focus groups were used in consumer research to develop a social marketing program to promote walking and other moderate-intensity physical activities. METHODS: Two phases of focus groups were conducted. Phase 1 groups, which included both men and women, were asked to respond to questions that would guide the development of a social marketing program based on social marketing concepts. Phase 1 also determined the intervention's target audience, which was irregularly active women aged 35 to 54. Phase 2 groups, composed of members of the target audience, were asked to further define the product and discuss specific promotion strategies. RESULTS: Phase 1 participants determined that the program product, or target behavior, should be walking. In addition, they identified price, place, and promotion strategies. Phase 2 participants determined that moderate-intensity physical activity is best promoted using the term exercise and offered suggestions for marketing walking, or exercise, to the target audience. CONCLUSION: There have been few published studies of social marketing campaigns to promote physical activity. In this study, focus groups were key to understanding the target audience in a way that would not have been accomplished with quantitative data alone. The group discussions generated important insights into values and motivations that affect consumers' decisions to adopt a product or behavior. The focus group results guided the development of a social marketing program to promote physical activity in the target audience in Sumter County, South Carolina. PMID- 16356362 TI - Race, ethnicity, and linguistic isolation as determinants of participation in public health surveillance surveys. AB - INTRODUCTION: To plan, implement, and evaluate programs designed to improve health conditions among racial and ethnic minority populations in the United States, public health officials and researchers require valid and reliable health surveillance data. Monitoring chronic disease and behavioral risk factors among such populations, however, is challenging. This study assesses the effects of race, ethnicity, and linguistic isolation on rates of participation in the Behavioral Risk Factor Surveillance System (BRFSS). METHODS: County-level data from the 2003 BRFSS survey and 2000 U.S. census were used to examine the effects of race, ethnicity, and linguistic isolation on six measures of survey participation (i.e., rates of resolution, screening, cooperation, response, language barriers, and refusal). RESULTS: Participation rates were significantly lower in counties with higher percentages of black people and people who did not speak English. Response rates decreased by 4.6% in counties with the highest concentration of black residents compared with counties with few black residents. Likewise, response rates decreased by approximately 7% in counties in which a larger percentage of the population spoke only Spanish or another Indo-European language compared with counties in which all residents spoke English. CONCLUSION: The negative relationship between the percentage of Spanish-only-speaking households and participation rates is troubling given that the BRFSS is conducted in both Spanish and English. The findings also indicate that more needs to be done to improve participation among other minorities. Researchers are investigating several ways of addressing disparities in participation rates, such as using postsurvey adjustments, developing more culturally appropriate data collection procedures, and offering surveys in multiple languages. PMID- 16356363 TI - Identifying geographic disparities in the early detection of breast cancer using a geographic information system. AB - INTRODUCTION: Identifying communities with lower rates of mammography screening is a critical step to providing targeted screening programs; however, population based data necessary for identifying these geographic areas are limited. This study presents methods to identify geographic disparities in the early detection of breast cancer. METHODS: Data for all women residing in Dane County, Wisconsin, at the time of their breast cancer diagnosis from 1981 through 2000 (N = 4769) were obtained from the Wisconsin Cancer Reporting System (Wisconsin's tumor registry) by ZIP code of residence. Hierarchical logistic regression models for disease mapping were used to identify geographic differences in the early detection of breast cancer. RESULTS: The percentage of breast cancer cases diagnosed in situ (excluding lobular carcinoma in situ) increased from 1.3% in 1981 to 11.9% in 2000. This increase, reflecting increasing mammography use, occurred sooner in Dane County than in Wisconsin as a whole. From 1981 through 1985, the proportion of breast cancer diagnosed in situ in Dane county was universally low (2%-3%). From 1986 through 1990, urban and suburban ZIP codes had significantly higher rates (10%) compared with rural ZIP codes (5%). From 1991 through 1995, mammography screening had increased in rural ZIP codes (7% of breast cancer diagnosed in situ). From 1996 through 2000, mammography use was fairly homogeneous across the entire county (13%-14% of breast cancer diagnosed in situ). CONCLUSION: The percentage of breast cancer cases diagnosed in situ increased in the state and in all areas of Dane County from 1981 through 2000. Visual display of the geographic differences in the early detection of breast cancer demonstrates the diffusion of mammography use across the county over the 20-year period. PMID- 16356364 TI - Evaluating progress toward Healthy People 2010 national diabetes objectives. AB - INTRODUCTION: In 1999, the National Diabetes Prevention and Control Program at the Centers for Disease Control and Prevention and its 59 Diabetes Prevention and Control Programs adopted five Healthy People 2010 objectives. These objectives aim to improve the rates of preventive care services among people with diabetes and include annual foot examinations, hemoglobin A1c tests, and annual dilated eye examinations. This paper examines progress toward meeting these three objectives. METHODS: Questions from the diabetes module of the Behavioral Risk Factor Surveillance System (BRFSS) were used to evaluate changes in age-adjusted rates for annual foot examinations, hemoglobin A1c tests, and annual dilated eye examinations of 44 jurisdictions between 2000 and 2003. Questions from the diabetes module were also used to compare percentage rates of 47 jurisdictions in 2003 with Healthy People 2010 percentage targets. RESULTS: From 2000 to 2003, for the 44 jurisdictions, the aggregate, age-adjusted rate of annual foot examinations increased from 63.7% to 69.3% (P < .001); the rate of self-reported hemoglobin A1c tests increased from 68.3% to 69.5% (P = .35); and the rate of annual dilated eye examinations decreased from 67.7% to 65.2% (P = .05). In 2003, 20 of 47 jurisdictions met the Healthy People 2010 target for foot or eye examinations, and all 47 jurisdictions met the target for hemoglobin A1c tests. An inverse association was found between baseline rates in 2000 and the magnitude of change from 2000 to 2003 for all three national diabetes objectives. CONCLUSION: The National Diabetes Prevention and Control Program should consider adopting additional Healthy People 2010 objectives. Baseline rates should be considered in 1) selecting objectives, 2) setting percentage targets, and 3) evaluating current or future objectives. Program-related information should be linked with traditional data sources such as BRFSS so that we can understand the role of environmental factors and evaluate progress of jurisdictions toward national diabetes objectives. PMID- 16356365 TI - A cost evaluation of the Georgia Stroke and Heart Attack Prevention Program. AB - INTRODUCTION: Hypertension is a leading cause of stroke, coronary artery disease, heart attack, and heart and kidney failure in the United States, all of which contribute to the rising costs of health care. The Georgia Stroke and Heart Attack Prevention Program is an education and direct service program for low income patients with hypertension. This project evaluated the cost-effectiveness of the program compared with the following two alternative scenarios: no treatment for high blood pressure and the typical hypertension treatment received in the private sector nationwide (usual care). METHODS: We estimated the preventive treatment costs and number of adverse health events averted (hemorrhagic and ischemic stroke, heart disease, and kidney failure) associated with the Georgia Stroke and Heart Attack Prevention Program in two Georgia health districts. We used program cost and service usage data obtained from the Georgia Department of Human Resources and probabilities and costs of expected adverse events published in peer-reviewed sources. We compared program costs and number of expected adverse health events averted with those expected from 1) no preventive care and 2) usual care for high blood pressure. RESULTS: The Georgia Stroke and Heart Attack Prevention Program was less costly and resulted in better health outcomes than either no preventive care or usual care. Compared with no preventive care in the two districts, the program was estimated to result in 54% fewer expected adverse events; compared with usual care, the program was estimated to result in 46% fewer expected adverse events. Combining the costs of preventive treatment with the costs of expected adverse events, the Georgia Stroke and Heart Attack Prevention Program cost an average of 486 dollars per patient annually, compared with average annual costs of 534 dollars for no care and 624 dollars for usual care. CONCLUSION: Maintaining a publicly financed stroke and heart attack prevention program is more cost-effective and results in greater health benefits than other plausible scenarios. Because the benefits of this program accrue to both the state and federal governments through reduced Medicaid and indigent care expenditures, both the state and federal governments have a financial incentive to support the program. PMID- 16356366 TI - The challenge of preventing cardiovascular disease in Tunisia. AB - Chronic disease, and particularly cardiovascular disease (CVD), is the major cause of death in most developed countries, despite the downward trend observed during the last three decades. Although CVD is emerging in developing countries, little is known there about comprehensive preventive measures for controlling its expansion. The health care system in Tunisia faces the challenge of increasing rates of CVD risk factors. Epidemiologic studies show high levels of CVD risk factors among Tunisian adults and children. Evidence shows that several risk factors and conditions are commonly associated with major chronic diseases. Integrated actions against selected risk factors (i.e., smoking, physical inactivity, and unhealthy diet), implemented within the social context, can lead to the reduction of major chronic diseases. These interventions should take place early in childhood. In Tunisia, a much-needed community-based intervention program to control CVD is being planned. This program will promote healthy living, smoke-free air, healthy nutrition, regular physical activity, and supportive living and working environments. Its ultimate goal is to reduce the burden of CVD and its related behaviors. A description of this program and how it will be implemented and assessed in the region of Sousse, Tunisia, is presented. PMID- 16356367 TI - Process, rationale, and interventions of Pakistan's National Action Plan on Chronic Diseases. AB - Most developing countries do not comprehensively address chronic diseases as part of their health agendas because of lack of resources, limited capacity within the health system, and the threat that the institution of national-level programs will weaken local health systems and compete with other health issues. An integrated partnership-based approach, however, could obviate some of these obstacles. In Pakistan, a tripartite public-private partnership was developed among the Ministry of Health, the nongovernmental organization (NGO) Heartfile, and World Health Organization. This was the first time an NGO participated in a national health program; NGOs typically assume a contractual role. The partnership developed a national integrated plan for health promotion and the prevention and control of noncommunicable diseases (NCDs), which as of January 2006 is in the first stage of implementation. This plan, called the National Action Plan on NCD Prevention, Control, and Health Promotion (NAP-NCD), was released on May 12, 2004, and attempts to obviate the challenges associated with addressing chronic diseases in countries with limited resources. By developing an integrated approach to chronic diseases at several levels, capitalizing on the strengths of partnerships, building on existing efforts, and focusing primary health care on chronic disease prevention, the NAP-NCD aims to mitigate the effects of national-level programs on local resources. The impact of the NAP-NCD on population outcomes can only be assessed over time. However, this article details the plan's process, its perceived merits, and its limitations in addition to discussing challenges with its implementation, highlighting the value of such partnerships in facilitating the missions and mandates of participating agencies, and suggesting options for generalizability. PMID- 16356368 TI - Barriers to diabetes self-management education programs in underserved rural Arkansas: implications for program evaluation. AB - BACKGROUND: Diabetes prevalence has reached epidemic proportions. Diabetes self management education (DSME) has been shown to improve preventive care practices and clinical outcomes. In this study, we discuss the barriers faced during the implementation of DSME programs in medically underserved rural areas of Arkansas. CONTEXT: Arkansas is a rural state, with most southeastern counties experiencing a shortage of health care professionals. The Arkansas Diabetes Prevention and Control Program and its partners established 12 DSME programs in underserved counties with a high prevalence of diabetes. METHODS: DSME programs were delivered by a registered nurse and a dietitian who provided 10 to 13 hours of education to each program participant. Baseline, 6-month, and year-end data were collected on preventive care practices, such as daily blood glucose monitoring, foot examination, systolic and diastolic blood pressure, and glycosylated hemoglobin level, among the participants in newly established DSME programs. CONSEQUENCES: Of the 12 DSME programs established, 11 received American Diabetes Association recognition. The number of participants in the DSME programs increased 138% in 1 year, from 308 in February 2003 to 734 in March 2004. Preventive care practices improved: daily blood glucose monitoring increased from 56% to 67% of participants, and daily foot examinations increased from 63% to 84% of participants. Glycosylated hemoglobin decreased by an average of 0.5 units per participant who completed the program. However, many anticipated and a few unanticipated barriers during the implementation of the program could not be overcome because of the lack of an evaluation plan. INTERPRETATION: Although results point to potential benefits of preventive care practices among DSME participants, interpretation of findings was limited by sample size. Sample size limitations are traced to barriers to assessing program outcome. Program evaluation should be integrated into the planning phase to ensure adequate measures of program effectiveness. PMID- 16356369 TI - A community coalition board creates a set of values for community-based research. AB - BACKGROUND: Researchers generally agree that communities should participate in the community-based research process, but neither a universally accepted approach to community participation nor a set of guiding principles exists. CONTEXT: The Morehouse School of Medicine Prevention Research Center was established in 1999 with the support of a grant from the Centers for Disease Control and Prevention. Its partners include a low-income, predominantly African American community, six public agencies, and two other academic institutions. A Community Coalition Board was established to represent the partners. The majority of the board is community members; it serves in a governance rather than an advisory capacity, with the community acting as the senior partner in interactions with the medical school, the agencies, and other academic institutions. METHODS: The Community Coalition Board developed a set of research priorities and a set of 10 community values, or principles, to guide research. A board committee reviews each protocol to ensure they uphold the values. CONSEQUENCES: The Community Coalition Board has been using the values since 1999, and in this article we describe its experience. After an initial period that included some disagreements between researchers and community members on the board, relationships have been good, and protocols have been approved with only minor changes. INTERPRETATION: Although the established community values reflect universally acknowledged principles of research ethics, they also address local concerns. An equal partnership between community members and researchers is most beneficial if the partners can agree on a set of values to govern research. PMID- 16356370 TI - The Prevention Research Centers Healthy Aging Research Network. AB - BACKGROUND: The Prevention Research Centers Healthy Aging Research Network (PRC HAN), funded by the Centers for Disease Control and Prevention's (CDC's) Healthy Aging program, was created in 2001 to help develop partnerships and create a research agenda that promotes healthy aging. The nine universities that participate in the network use their expertise in aging research to collaborate with their communities and other partners to develop and implement health promotion interventions for older adults at the individual, organizational, environmental, and policy levels. CONTEXT: The population of older adults in the United States is growing rapidly; approximately 20% of Americans will be aged 65 years or older by 2030. The health and economic impact of an aging society compel the CDC and the public health community to place increased emphasis on preventing unnecessary disease, disability, and injury among older Americans. METHODS: The PRC-HAN has a broad research agenda that addresses health-promoting skills and behaviors, disease and syndrome topics, and knowledge domains. The network chose physical activity for older adults as its initial focus for research and has initiated two networkwide projects: a comprehensive, multisite survey that collected information on the capacity, content, and accessibility of physical activity programs for older adults and a peer-reviewed publication that describes the role of public health in promoting physical activity among older adults. In addition to participating in the core research area, each network member works independently with its community committee on PRC-HAN activities. CONSEQUENCES: As a result, the network is 1) expanding prevention research for older adults and their communities; 2) promoting the translation and dissemination of findings to key stakeholders; 3) strengthening PRC-HAN capacity through partnerships and expanded funding; and 4) stimulating the adoption of policies and programs by engaging policymakers, planners, and practitioners. In 2003, the PRC-HAN initiated an internal evaluation to better define the network's contributions to healthy aging, formalize internal processes, and better equip itself to serve as a model for other PRC thematic networks. The PRC-HAN is conducting a pilot evaluation for eventual inclusion in the PRC national evaluation. INTERPRETATION: The PRC-HAN has established itself as an effective research network to promote healthy aging. It has developed trust and mutual respect among participants, forged strong ties to local communities, and shown the ability to combine its expertise in healthy aging with that of partners in national, state, and local organizations. PMID- 16356372 TI - Steps to a HealthierUS Cooperative Agreement Program: foundational elements for program evaluation planning, implementation, and use of findings. AB - The Steps to a HealthierUS Cooperative Agreement Program (Steps Program) enables funded communities to implement chronic disease prevention and health promotion efforts to reduce the burden of diabetes, obesity, asthma, and related risk factors. At both the national and community levels, investment in surveillance and program evaluation is substantial. Public health practitioners engaged in program evaluation planning often identify desired outcomes, related indicators, and data collection methods but may pay only limited attention to an overarching vision for program evaluation among participating sites. We developed a set of foundational elements to provide a vision of program evaluation that informs the technical decisions made throughout the evaluation process. Given the diversity of activities across the Steps Program and the need for coordination between national- and community-level evaluation efforts, our recommendations to guide program evaluation practice are explicit yet leave room for site-specific context and needs. Staff across the Steps Program must consider these foundational elements to prepare a formal plan for program evaluation. Attention to each element moves the Steps Program closer to well-designed and complementary plans for program evaluation at the national, state, and community levels. PMID- 16356371 TI - The Annual African American Conference on Diabetes: evolving program evaluation with evolving program implementation. AB - BACKGROUND: According to 2003 Behavioral Risk Factor Surveillance System data, South Carolina has the fourth highest rate of overall diabetes among the 50 states (9.3%) but the second highest rate among African Americans (15.5%). Nationwide, African Americans are disproportionately affected by diabetes. In addition, 40% of the African American population in South Carolina lives in a rural area, and approximately 26% live at or below the poverty level. Lack of access to health care and diabetes education are additional barriers for people with diabetes and their families. CONTEXT: Since 1997, the South Carolina Diabetes Prevention and Control Program and the Diabetes Today Advisory Council have sponsored the African American Conference on Diabetes, which targets African Americans with diabetes, their families, and their caregivers. This article describes the evolution of the conference and its evaluation. METHODS: In 2002, we conducted focus groups with 20 African American conference attendees with diabetes to 1) assess the program's effects, 2) determine how to reach more individuals, and 3) improve programming. In 2004, we incorporated the preconference and postconference Diabetes Understanding Scale survey to assess the cognitive impact of the conference on participants. CONSEQUENCES: Focus group results revealed that participants wanted to attend the conference because of the opportunity to increase their knowledge and change their behaviors through 1) education, 2) social support, 3) resources, and 4) logistics. Self-rated understanding increased significantly after the conference for each cognitive understanding item on the Diabetes Understanding Scale. INTERPRETATION: Focus group results suggested that participants who continue to attend the conference year after year may improve diabetes self-management skills. A quantitative evaluation showed that this 1-day diabetes education conference significantly increased short-term, self-rated cognitive understanding of diabetes behaviors. PMID- 16356373 TI - Development of the Diabetes Indicators and Data Sources Internet Tool (DIDIT). AB - Developing a Web-based tool that involves the input, buy-in, and collaboration of multiple stakeholders and contractors is a complex process. Several elements facilitated the development of the Web-based Diabetes Indicators and Data Sources Internet Tool (DIDIT). The DIDIT is designed to enhance the ability of staff within the state-based Diabetes Prevention and Control Programs (DPCPs) and the Centers for Disease Control and Prevention (CDC) to perform diabetes surveillance. It contains information on 38 diabetes indicators (measures of health or factors associated with health) and 12 national- and state-level data sources. Developing the DIDIT required one contractor to conduct research on content for diabetes indicators and data sources and another contractor to develop the Web-based application to house and manage the information. During 3 years, a work group composed of representatives from the DPCPs and the Division of Diabetes Translation (DDT) at the CDC guided the development process by 1) gathering information on and communicating the needs of users and their vision for the DIDIT, 2) reviewing and approving content, and 3) providing input into the design and system functions. Strong leadership and vision of the project lead, clear communication and collaboration among all team members, and a commitment from the management of the DDT were essential elements in developing and implementing the DIDIT. Expertise in diabetes surveillance and software development, enthusiasm, and dedication were also instrumental in developing the DIDIT. PMID- 16356374 TI - The REACH 2010 logic model: an illustration of expected performance. AB - The Centers for Disease Control and Prevention (CDC) supports 40 Racial and Ethnic Approaches to Community Health (REACH 2010) community coalitions in designing, implementing, and evaluating community-driven strategies to eliminate health disparities in racial and ethnic groups. The REACH 2010 logic model was developed to assist grantees in identifying, documenting, and evaluating local attributes of the coalition and its partners to reduce and eliminate local health disparities. The model emphasizes the program's theory of change for addressing health disparities; it displays five distinct stages of evaluation for which qualitative and quantitative measurement data are collected. The CDC is relying on REACH 2010 grantees to provide credible evidence that explains how community contributions have changed conditions and behaviors, thus leading to the reduction and elimination of health disparities. PMID- 16356375 TI - Transforming a Master of Public Health program to address public health practice needs. AB - The future of the constantly changing public health profession is tied to the development of practice skills through competency-based training. In this article, we describe a program change in the Master of Public Health program at East Stroudsburg University in northeastern Pennsylvania. The first goal of the program transition was to ensure that all program elements included the relevant vision, values, mission, goals, and objectives. The second goal was to use continuous data input and evaluation to incorporate opportunities for flexible assessments. The change process helped the university faculty define the program's vision and fostered an environment of community collaboration that guides training for public health professionals. PMID- 16356376 TI - The Nutrition and Physical Activity Program to Prevent Obesity and Other Chronic Diseases: monitoring progress in funded states. AB - To help address the challenges posed by the obesity epidemic in the United States, the U.S. Congress authorized the Centers for Disease Control and Prevention to establish the Nutrition and Physical Activity Program to Prevent Obesity and Other Chronic Diseases. In this article, we summarize the progress of the first 20 states funded by this program. The data presented are based on the information provided by the states in their semiannual progress monitoring reports on program activities from January through June 2004. The states have made progress in developing capacity and infrastructure for their programs, including leveraging financial resources and developing strong partnerships. In addition, they are planning and initiating environmental changes through legislation, and, although less frequently, through policies and other changes such as expanding physical activity opportunities. Collectively, the states are making progress in planning and implementing activities to prevent and control obesity and other chronic diseases. PMID- 16356377 TI - A six-step model for evaluation of community-based physical activity programs. PMID- 16356378 TI - Solution for survey discrepancies in Washington State smoking prevalence. PMID- 16356379 TI - Provision of school-based preventive oral health services to Medicaid beneficiaries. PMID- 16356380 TI - Response to "Old black water". PMID- 16356381 TI - A lemming's view of rural practice. PMID- 16356382 TI - President's message. Summertime and the livin' ain't easy. PMID- 16356383 TI - Qualitative troponin I estimation in the diagnosis of acute coronary syndromes in three rural hospitals. AB - OBJECTIVE: To examine the utility of point-of-care qualitative troponin I (TnI) testing in patients with possible acute coronary syndromes (ACS). METHODS: A retrospective chart review of all patients undergoing qualitative TnI testing between September 2001 and February 2002 was conducted at the emergency departments of 3 rural hospitals in Alberta. We looked at the incidence of ACS, the comparison between TnI and creatine kinase (CK) testing and the timing of testing. RESULTS: Of the 235 patients tested, 8 had ST-elevation myocardial infarctions and 11 non ST-elevation infarctions. One patient had unstable angina with minimal myocardial damage. Qualitative TnI testing was positive in all 14 cases of infarction tested more than 6 hours after symptom onset, and CK elevation occurred in 15/17 cases (TnI sensitivity 1.0 [95% confidence interval (CI) 0.78-1.0], CK sensitivity 0.882 [95% CI 0.66-0.97]). There were 3 positive TnI tests and 33 raised CK levels in patients without evidence for ACS (TnI specificity 0.986 [95% CI 0.96-0.99], likelihood ratio [LR] 72.0 [95% CI 23.4 221.5]); CK specificity 0.847 [95% CI 0.79-0.89], LR 5.8 [95% CI 4.0-8.3]). In 44 patients (20.8%) TnI testing was inappropriately not repeated more than 6 hours after symptom onset. CONCLUSION: Qualitative TnI testing appears highly sensitive and more specific than CK estimation in detecting myocardial infarction. Diagnostic algorithms must emphasize the importance of testing 6 or more hours after symptom onset. PMID- 16356384 TI - Relationship between practice location of Ontario family physicians and their rural background or amount of rural medical education experience. AB - INTRODUCTION: The present study was designed to determine if there was a difference in rural background and rural medical education experience between practising rural physicians and practising urban physicians in Ontario. METHOD: A cross-sectional survey was mailed to 507 strictly defined rural family physicians and 505 urban family physicians practising in Ontario. The main outcome measures were population of the community while growing up, rural medical education and medical school attended. RESULTS: Responses of 264 rural physicians were compared with 179 urban physician responses. The groups were comparable in years of practice. Rural physicians were significantly more likely to have grown up in a rural community (34.9% v. 14.6%), to have had clinical training in a rural setting during medical school (55.4% v. 35.2%) and to have had clinical training in a rural setting of 8 weeks or more during postgraduate residency training (38.8% v. 20.2%). During residency training, longer duration of rural placements (more than 6 months) was significantly associated with practice in a rural area (15.5% of rural physicians, 1.7% of urban physicians). After controlling for other predictors, each of the following were independent variables: growing up in a community of less than 10 000 people (odds ratio [OR] 3.31), having had some undergraduate rural clinical training (OR 2.46), having had postgraduate rural training of 8 weeks or more (OR 2.17), attending a Canadian medical school outside Ontario (OR 3.80) and being male (OR 2.57). CONCLUSION: Practising rural physicians compared with urban physicians were significantly more likely to have come from a rural background, to have had an undergraduate rural medical education, to have had postgraduate rural training, to have graduated from a Canadian medical school outside Ontario, and to be male. Each of these had an independent effect on practice location. PMID- 16356385 TI - Women's health in northern British Columbia: the role of geography and gender. AB - INTRODUCTION: Although research interest in women's health is growing, much of the literature does not sufficiently describe the importance of geography and gender for the health of women. This qualitative study explored factors in the northern Canadian context that influence women's health by interviewing 25 women in northern Canada. RESULTS: Findings reveal that the importance of the northern context for women's health can be attributed to the north's historical location, and its physical, sociocultural and political environments. The northern context contributes to the marginalization of northern women that is characterized by isolation, limited options, limited power and being silenced. CONCLUSION: Health care practice and policy must attend to contextual as well as individual and sociocultural factors if women's health is to be advanced in northern settings. PMID- 16356386 TI - The occasional removal of an embedded fish hook. PMID- 16356387 TI - Country cardiograms case 28. Ventricular hypertrophy. PMID- 16356388 TI - Polar physician. PMID- 16356389 TI - CMA Web site: InfoPOEMs. PMID- 16356390 TI - The diversity of Family Medicine Fellowship programs. PMID- 16356391 TI - TP53, Bcl-2 and growth hormone receptor expression in cutaneous squamous cell carcinoma. AB - The incidence and prevalence of premalignant and malignant skin lesions including squamous cell carcinoma (SCC) of the skin are increasing worldwide. The aim of the study was to determine TP53, Bcl-2 and growth hormone receptor (GHR) expression in SCC and to investigate relative importance of these proto-oncogenes in its biological behavior. Expression of TP53, Bcl-2 and GHR was determined by immunohistochemistry in 27 SCC specimens and adjacent perilesional skin. The relative proportion of immunoreactive cells was counted with semiquantitative method. TP53 positivity was detected in 24 (89%), Bcl-2 in 18 (67%) and GHR in 25 (93%) of 27 SCC specimens investigated. In comparison with perilesional skin, TP53 and GHR positivity was significantly increased, and Bcl-2 positivity significantly decreased in SCC. Increased TP53 expression in SCC lesions implies that Tp53 mutation is an early and crucial event in its development. Increased GHR expression suggests a role of growth hormone in the development of SCC. PMID- 16356392 TI - Expression of keratins 14, 10 and 16 in marginal keratoderma of the palms. AB - Marginal keratoderma of the palms (MKP), also known as degenerative collagenous plaques of the hands, is an infrequent disease affecting individuals with photoaged skin and submitted to mechanical trauma of the hands. The aim of the study was to study the expression of keratin 14 (K14) in the basal layer and keratins 10 (K10) and 16 (K16) in the suprabasal layer, and to establish the effect of growth factors and receptors of the epidermal growth factor in the genesis of hyperkeratosis observed in MKP. The study included 14 patients with MKP. Expression of keratins 10, 14 and 16 was visualized by immunohistochemical staining using monoclonal antibodies by standard immunoperoxidase method. As control three normal skin samples were used. Absence or weak expression of keratins 14 and 10, and strong expression of keratin 16 was recorded in MKP patients. It was concluded that MKP patients present weak expression of keratins 14 (basal membrane) and 10 (suprabasal layer) but strong expression of keratin 16, emphasizing the importance of stimulation of epidermal growth factor receptors by ultraviolet radiation and traumatism. PMID- 16356393 TI - Surgical treatment of axillary hyperhidrosis with liposuction equipment: risks and benefits. AB - Axillary hyperhidrosis poses a serious problem to the affected patients. So far, the conservative measures employed seem to be disappointing, operations with axillary skin excision, undermining and/or skin reconstruction may cause secondary functional and cosmetic problems, whereas botulinum A toxin injections need to be repeated frequently. The aim of this study was to establish the safety, efficacy, and durability of subdermal (subcorial) suction sweat gland curettage in the treatment of axillary hyperhidrosis. So far, the method seems to be devoid of possible risks and drawbacks. In the last 4.5 years, 15 patients with axillary hyperhidrosis were operated on with the use of liposuction tools. First operations were performed in general anesthesia, then in tumescent anesthesia. The procedure of suction curettage was performed with the use of 3- to 4-mm wide liposuction cannulas. The patients were closely monitored during early stages of the healing process; then they were evaluated at 1 and 3 months, and finally at 1-4 years of the operation, when they were asked to assess the effects of the operation. Four patients had recurrence of the disease within 3 months; three of them were reoperated on, with good result. At 1-4 years of the operation, all our responders (ten of 15 patients) stated that the disease had completely subsided. The following complications were observed during the process of healing: hematomas, transient skin unevenness, and partial skin flap necrosis. In conclusion, subdermal suction curettage seems to be superior to botulinum A toxin injections by the effect durability, and to the surgical methods with skin excision and undermining by the probably lower complication rates. PMID- 16356394 TI - The new scoring system for evaluation of skin inflammation extent and severity in patients with atopic dermatitis. AB - The new scoring system for assessment of the extent and severity of skin inflammation index in atopic dermatitis patients, W-AZS, is presented. The system provides detailed assessment of both subjective and objective signs and symptoms of atopic dermatitis. With the use of W-AZS, acute and chronic skin manifestations of inflammatory process are appropriately evaluated and scored. It also enables the practitioner to assess various localizations of skin lesions at different time points. W-AZS is a relatively easy and rapid index to perform, and it seems very beneficial for clinicians. Other scoring systems used in atopic dermatitis are also presented, analyzed and compared, e.g., Atopic Dermatitis Area and Severity Index (ADASI), SCORing Atopic Dermatitis (SCORAD), Eczema Area and Severity Index (EASI), Six Area, Six Sign Atopic Dermatitis (SASSAD), and Three-Item Severity score (TIS). There is a strong necessity to standardize clinical evaluation of the extent and severity of skin diseases such as atopic dermatitis, as laboratory techniques and parameters are not really of great use for practitioners. PMID- 16356395 TI - Epicutaneous patch test reactions in atopic dermatitis patients. AB - During the 1998-2003 period, patch testing was carried out in 65 atopic dermatitis patients, 20 (31%) male and 45 (69%) female, mean age 34.7 (range 6 77) years. Twenty-six (40%) patients, 7 (27%) male and 19 (73%) female, showed positive reaction to one or more allergens. Allergic reactions were more common in women. The most common allergens were nickel (33.3%), cobalt (11.1%), fragrance mix (11.1%), white mercury precipitate (8.4%), and others (36.1%). There was a significant association between nickel allergy and cobalt allergy. It is concluded that contact hypersensitivity is not rare in patients with atopic dermatitis. PMID- 16356396 TI - Epidemiology of acquired bullous diseases in Eastern Croatia: a retrospective prewar to postwar study. AB - The purpose of this study was to evaluate the epidemiology of bullous diseases (BD) in eastern Croatia during a ten-year period (1986-1990/1992-1996), and to estimate the effect of prolonged exposure to traumatic events during the war in Croatia on the prevalence and incidence of acquired BD. The files of all BD patients hospitalized at Department of Dermatology and Venereology, Osijek University Hospital, during the periods from January 1986 to December 1990 and from January 1992 to December 1996 were collected and analyzed with regard to personal data, history of the disease including age, sex and onset of symptoms, clinical diagnosis, laboratory findings, and associated illness. Forty-five patients were newly diagnosed with BD over the ten-year period. During the 1986 1990 period, 19 patients with BD represented 0.89% of 2133 patients admitted to our Department. During the 1992-1996 period, 26 newly diagnosed patients with BD represented 1.27% of 2050 patients treated at our Department. Females were more affected than males. The most common clinical variant was pemphigus vulgaris, occurring frequently in the middle-aged population. All our patients were exposed to prolonged stressful war conditions during the 1992-1996 period, therefore, we speculate that extended emotional stress may have triggered the onset of the disease. PMID- 16356397 TI - Atypical clinical appearance of pemphigus vulgaris on the face: case report. AB - Pemphigus vulgaris is an organ-specific autoimmune mucocutaneous disorder. In the majority of cases, the disease manifests initially with oral lesions, and may be limited to a single site for months before spreading. A 78-year-old woman with yellowish crusted areas on her left preauricular region and close to the medial angle of her right eye is presented. Although she described an episode of erosions on her lower lip, the involvement of mucosal surfaces was not noticed on examination. Before she presented to our Department, she was misdiagnosed as an actinic cheilitis and malignant skin tumor. Histopathologic examination and direct immunofluorescence confirmed the diagnosis of pemphigus vulgaris. Immunoblotting of epidermal extracts detected IgG antibodies against desmoglein 3 but not desmoglein 1, which was also confirmed by ELISA test. The patient responded favorably to systemic corticosteroid therapy combined with adjuvant immunosuppressive therapy, with complete clearance of the lesions. PMID- 16356398 TI - Herpes simplex: autoinoculation versus dissemination. AB - Autoinoculation and dissemination (or Kaposi's varicelliform eruption (KVE) or eczema herpeticum) of herpetic lesions are two forms of viral spread, and it is essential to differentiate the two. Autoinoculation means true infection with retrograde transport of the virus to the dorsal root ganglia of the relevant dermatome that allows the virus to remain there in a latent state for a lifetime, with periodic reactivation. Autoinoculation is, in a manner of speaking, a kind of self-infection with a virus that exists in the host. In contrast, KVE involves a spread of the lesions to the skin areas affected by another skin disease, but there is no true inoculation, i.e. the nerve endings and ganglion are not affected, and so reactivation and recurrences of these lesions will not usually occur. Four cases of autoinoculation and two of KVE illustrate the differences between these two forms of viral spread. PMID- 16356399 TI - The role of nitric oxide in the pathogenesis of venous ulcers. AB - Chronic venous insufficiency frequently leads to ulceration. The exact pathophysiological mechanisms underlying the development of venous ulceration remain to be elucidated. One major etiological factor of the trophic changes is the phenomenon of leukocyte trapping. The aim of the study was to review the pathophysiological events culminating in venous ulceration, focusing primarily on the role of alterations in nitric oxide (NO) production. We establish the hypothesis that venous stasis in the microcirculation reduces the rate of shear stress on the endothelial cells, effectively resulting in a decrease in cellular levels of NO, a key event of enhanced adhesion molecule expression and subsequent massive neutrophil activation. A similar series of events is proposed to explain the ischemic-reperfusion tissue injury. Inducible NO (iNO) produced by the inflammatory cells causes free radical injury seen as a venous ulceration. PMID- 16356400 TI - Health problems in musicians--a review. AB - Among artists, musicians suffer from special health problems related to their occupation and lifestyle. These mainly include musculoskeletal disorders, skin disorders, respiratory disorders, and noise induced hearing impairment. In this review we point out the most prevalent health problems that musicians face in their professional lives as well as the means to prevent them. The characteristics of individual performer and the type of instrument that he/she plays will determine the occupational disorder or disease that a musician may suffer from. The main worry about health problems in musicians is that they can become potentially career-ending disabilities. Therefore well-planned preventive strategies should be employed to help musicians in obtaining long and healthy career. PMID- 16356401 TI - What is evidence-based dermatology? PMID- 16356415 TI - Lost in the black box: juvenile depression, suicide, and the FDA's black box. PMID- 16356416 TI - The relationship between ethnicity and outcomes in solid organ transplantation. PMID- 16356417 TI - Down syndrome: immunologic and epidemiologic associations-enigmas remain. PMID- 16356418 TI - Secular trends in menarche. PMID- 16356419 TI - Will, ideas, and execution: their role in reducing adverse medication events. PMID- 16356420 TI - The expanding spectrum of congenital disorders of glycosylation. PMID- 16356421 TI - The importance of calcium-binding proteins in childhood diseases. PMID- 16356422 TI - Disparities in outcome for black patients after pediatric heart transplantation. AB - OBJECTIVE: To examine the relationship of black race to graft survival after heart transplantation in children. STUDY DESIGN: United Network for Organ Sharing records of heart transplantation for subjects <18 years of age from 1987 to 2004 were reviewed. Analysis was performed using proportional hazards regression controlling for other potential risk factors. RESULTS: Of the 4227 pediatric heart transplant recipients, 717 (17%) were black. The 1-year graft survival rate did not differ among groups; however, the 5-year graft survival rate was significantly lower for black recipients, 51% versus 69%, P < .001. The median graft survival for black recipients was 5.3 years as compared with 11.0 years for other recipients. Black recipients had a greater number of human leukocyte antigen mismatches, lower median household income, and a greater percentage with Medicaid as primary insurance, P < .001, P < .001, and P < .001. After adjusting for economic disparities, black race remained significantly associated with graft failure, odds ratio = 1.67 (95% CI 1.47 to 1.87), P < .001. CONCLUSIONS: Median graft survival after pediatric heart transplantation for black recipients is less than half that of other racial groups. These differences do not appear to be related primarily to economic disparities. PMID- 16356423 TI - Intrinsic abnormalities of lymphocyte counts in children with down syndrome. AB - OBJECTIVE: Down syndrome (DS) is associated with an increased frequency of infections, hematologic malignancies, and autoimmune diseases, suggesting that immunodeficiency is an integral part of DS that contributes significantly to the observed increased morbidity and mortality. We determined the absolute counts of the main lymphocyte populations in a large group of DS children to gain further insight into this immunodeficiency. STUDY DESIGN: In a large group of children with DS (n = 96), the absolute numbers of the main lymphocyte subpopulations were determined with 3-color immunophenotyping using the lysed whole-blood method. The results were compared with previously published data in healthy children without DS. RESULTS: In healthy children with DS, the primary expansion of T and B lymphocytes seen in healthy children without DS in the first years of life was severely abrogated. The T- lymphocyte subpopulation counts gradually reached more normal levels with time, whereas the B- lymphocyte population remained severely decreased, with 88% of values falling below the 10th percentile and 61% below the 5th percentile of normal. CONCLUSIONS: The diminished expansion of T and B lymphocytes strongly suggests that a disturbance in the adaptive immune system is intrinsically present in DS and is not a reflection of precocious aging. Thymic alterations have been described in DS that could explain the decreased numbers of T lymphocytes, but not the striking B lymphocytopenia, seen in these children. PMID- 16356424 TI - Risk of death for children with down syndrome and sepsis. AB - OBJECTIVE: To determine differences in case fatality rates between children with and without Down syndrome. STUDY DESIGN: We used the Pediatric Health Information System (PHIS) database, which includes demographic and diagnostic data from freestanding children's hospitals. Using Poisson regression, we determined the risk of mortality from sepsis for children with Down syndrome, after controlling for potential confounding factors. RESULTS: A total of 35,645 patients met our inclusion criteria, of which 3936 (11%) died during hospitalization. Altogether, 620 of the included patients also had a diagnosis of Down syndrome; 106 (17%) of these died during hospitalization. Children with Down syndrome had significantly elevated risk of mortality (mortality rate ratio = 1.30; 95% confidence interval = 1.06 to 1.59) after adjusting for potential confounding factors including demographics, pathogens, and concomitant conditions. CONCLUSIONS: Children with Down syndrome and sepsis have elevated risk of mortality. These findings have implications for treatment decisions, communications about prognosis, and future research. PMID- 16356426 TI - Interpreting the continued decline in the average age at menarche: results from two nationally representative surveys of U.S. girls studied 10 years apart. AB - OBJECTIVES: To determine whether average age at menarche declined in the United States during the past decade, and whether associations between menarcheal timing, weight status, and race/ethnicity changed. STUDY DESIGN: Relative weight, race/ethnicity, and menarcheal status of girls (n = 1577) in the National Health and Nutrition Examination Survey (NHANES) III (1988-1994) were compared with those of girls (n = 1720) in NHANES 1999-2002. Probit analysis estimated average age at menarche overall and also by race/ethnicity. Logistic regression assessed associations of relative weight and race/ethnicity with menarcheal status. RESULTS: In the United States, average age at menarche declined from 12.53 years (95% confidence interval [CI] = 12.43 to 12.63 years) in 1988-1994 to 12.34 years (95% CI = 12.24 to 12.45 years) in 1999-2002. By race/ethnicity, average age at menarche estimates were as follows: non-Hispanic whites, 12.57 years (95% CI = 12.45 to 12.69 years) and 12.52 years (95% CI = 12.38 to 12.67 years); non Hispanic blacks, 12.09 years (95% CI = 11.82 to 12.36 years) and 12.06 years (95% CI = 11.81 to 12.32 years); and Mexican Americans, 12.24 years (95% CI = 11.88 to 12.59 years) and 12.09 years (95% CI = 11.81 to 12.37 years). Higher relative weight was consistently associated with increased likelihood of having reached menarche. CONCLUSIONS: Average age at menarche in the United States declined by 2.3 months between 1988-1994 and 1999-2002; by race/ethnicity, declines were considerably smaller. Changes in the population distribution of race/ethnicity and relative weight should be considered when interpreting trends in age at menarche. PMID- 16356427 TI - Potential medication dosing errors in outpatient pediatrics. AB - OBJECTIVE: To determine the prevalence of potential dosing errors of medication dispensed to children for 22 common medications. STUDY DESIGN: Using automated pharmacy data from 3 health maintenance organizations (HMOs), we randomly selected up to 120 children with a new dispensing prescription for each drug of interest, giving 1933 study subjects. Errors were defined as potential overdoses or potential underdoses. Error rate in 2 HMOs that use paper prescriptions was compared with 1 HMO that uses an electronic prescription writer. RESULTS: Approximately 15% of children were dispensed a medication with a potential dosing error: 8% were potential overdoses and 7% were potential underdoses. Among children weighing <35 kg, only 67% of doses were dispensed within recommended dosing ranges, and more than 1% were dispensed at more than twice the recommended maximum dose. Analgesics were most likely to be potentially overdosed (15%), whereas antiepileptics were most likely potentially underdosed (20%). Potential error rates were not lower at the site with an electronic prescription writer. CONCLUSIONS: Potential medication dosing errors occur frequently in outpatient pediatrics. Studies on the clinical impact of these potential errors and effective error prevention strategies are needed. PMID- 16356429 TI - Influence of timing and dose of thyroid hormone replacement on mental, psychomotor, and behavioral development in children with congenital hypothyroidism. AB - OBJECTIVES: To evaluate the influence of initial and postinitial treatment factors on cognitive, psychomotor, and psychological outcome in schoolchildren with congenital hypothyroidism (CH). STUDY DESIGN: We studied 45 patients (19 with severe CH and 26 with mild CH) and 37 control children by correlating initial and postinitial treatment factors (free thyroxine and thyroid-stimulating hormone [TSH] concentrations, and the percentage of overtreatment and undertreatment periods) with the results of neuropsychological tests and behavior (as reported on the Teacher Report Form [TRF]). RESULTS: The global IQ of the children with CH was comparable to that of the controls; visuomotor and verbal scores were lower, and total TRF scores were higher. Ethnic group, previous development, and overtreatment predicted IQ and verbal scores, with higher scores seen for the overtreated patients than for the control children and those patients who had not been overtreated. As initial treatment was less satisfactory, total TRF scores were higher. CONCLUSIONS: Our study suggests that initial and postinitial suboptimal treatment of CH leads to abnormalities in IQ and specific fields. Overtreatment may advance cognitive development in 5-1/2- to 7-year-olds. Suboptimal initial treatment may lead to behavioral problems. We recommend that TSH concentrations be maintained within the normal range in patients with CH. PMID- 16356430 TI - Neurodevelopmental outcomes in congenital hypothyroidism: comparison of initial T4 dose and time to reach target T4 and TSH. AB - OBJECTIVES: To compare neurodevelopmental outcomes in severe and moderate congenital hypothyroidism (CH) among 3 different initial L-thyroxine doses and to examine the effect of the time to thyroid function normalization on neurodevelopmental outcomes. STUDY DESIGN: Neurodevelopmental assessments of 31 subjects included the Mullen Scales of Early Learning, Wechsler Preschool and Primary Scale of Intelligence-Revised, Wechsler Intelligence Scale for Children, Wide-Range Achievement Test, and Child Behavioral Checklist. RESULTS: Subjects started on higher initial L-thyroxine doses (50 mug) had full-scale IQ scores 11 points higher than those started on lower (37.5 mug) initial doses. However, verbal IQ, performance IQ, and achievement scores did not differ among the 3 treatment cohorts. Subjects with moderate CH had higher full-scale IQ scores than subjects with severe CH, regardless of the initial treatment dose. Subjects who took longer than 2 weeks to normalize thyroid function had significantly lower cognitive, attention, and achievement scores than those who achieved normal thyroid function at 1 or 2 weeks of therapy. CONCLUSIONS: Initial L-thyroxine dose and faster time to normalization of thyroid function are important to optimal neurodevelopmental outcome. In severe CH, it is important to choose an initial dose at the higher end of the recommended range to achieve these goals. PMID- 16356431 TI - A randomized controlled trial of fluid supplementation in term neonates with severe hyperbilirubinemia. AB - OBJECTIVE: To evaluate the effectiveness of fluid supplementation in decreasing the rate of exchange transfusion and the duration of phototherapy in term neonates with severe nonhemolytic hyperbilirubinemia. STUDY DESIGN: This was a randomized controlled trial conducted in a tertiary care referral unit in northern India. Seventy-four term neonates with severe nonhemolytic hyperbilirubinemia (total serum bilirubin > 18 mg/dL [308 micromol/L] to < 25 mg/dL [427 micromol/L]). The subjects were randomized to an "extra fluids" group (intravenous fluid supplementation for 8 hours and oral supplementation for the duration of phototherapy; n = 37) or a control group (n = 37). RESULTS: At inclusion, 54 infants (73%) had high serum osmolality, including 28 (75%) in the extra fluids group and 26 (70%) in the control group. The proportion of infants who underwent exchange transfusion was lower in the extra fluids group than in the control group: 6 (16%) versus 20 (54%)(P = .001; relative risk = 0.30; 95% confidence interval = 0.14 to 0.66). The duration of phototherapy was also shorter in the extra fluids group: 52 +/- 18 hours versus 73 +/- 31 hours (P = .004). CONCLUSION: Fluid supplementation in term neonates presenting with severe hyperbilirubinemia decreased the rate of exchange transfusion and duration of phototherapy. PMID- 16356432 TI - Association between fluid intake and weight loss during the first ten days of life and risk of bronchopulmonary dysplasia in extremely low birth weight infants. AB - OBJECTIVE: To demonstrate the association between fluid intake and weight loss during the first 10 days of life and the risk of bronchopulmonary dysplasia (BPD) in extremely low birth weight (ELBW) infants. STUDY DESIGN: A retrospective analysis of data from a cohort of ELBW infants enrolled in the Neonatal Research; 1,382 infants with birth weight between 401 and 1,000 g were randomized. The daily fluid intake and weight loss during the first 10 days of life were compared between the infants who survived without BPD and those who either died or developed BPD. Demographic and clinical neonatal variables were also compared. Multivariate logistic regression was used to analyze the effect of fluid intake and weight loss on death or BPD, controlling for demographic and clinical factors that are significantly associated with BPD by univariate analysis. RESULTS: 585 infants survived without BPD and 797 infants either died or developed BPD. Univariate analysis showed that the daily fluid intakes were higher (day 2-10) and weight loss less (day 6-9) in the group of infants who either died or developed BPD. In addition, lower birth weight, lower gestational age, male gender, lower 1 and 5-minute Apgar Scores, higher oxygen requirement at 24 hours of age, longer duration of assisted ventilation, use of postnatal steroids for BPD and presence of severe intraventricular hemorrhage, proven necrotizing enterocolitis, patent ductus arteriosus, and late onset sepsis, were associated with higher incidence of death or BPD. The adjusted risk of higher fluid intake and less weight loss during the first 10 days of life remained significantly related to death or BPD. CONCLUSION: In this cohort of ELBW infants treated during the post surfactant era, higher fluid intake and less weight loss during the first 10 days of life were associated with an increased risk of BPD. The finding suggests that careful attention to fluid balance might be an important means to reduce the incidence of BPD. PMID- 16356433 TI - Bone measurements by peripheral quantitative computed tomography (pQCT) in children with cerebral palsy. AB - OBJECTIVE: To use peripheral quantitative computed tomography (pQCT) to determine bone measurements in patients with cerebral palsy (CP) age 3 to 20 years and compare them with control subjects. STUDY DESIGN: A total of 13 (5 male) patients with CP, along with 2 sex- and age-matched controls for each, were included in a mixed-model analysis with matched pairs as random effects for pQCT bone measurements of the 20% distal tibia. RESULTS: Tibia length was similar in the CP and control groups (P = .57). Weight was marginally higher in the control group (P = .06). Cortical bone mineral content (BMC), area, thickness, polar strength strain index (pSSI), and periosteal and endosteal circumferences were greater in the control group (P < .05 for all). Relationships between bone measurements and weight showed that cortical BMC, area, periosteal circumference, and pSSI were greater at higher weights in the control group (group-by-weight interaction, P < .05 for all). Cortical thickness was greater in the control group and was correlated with weight. Cortical volumetric bone mineral density (vBMD) was greater with higher weights in the CP group (group-by-weight interaction, P = .03). CONCLUSIONS: Bone strength, as indicated by pSSI, is compromised in children with CP due to smaller and thinner bones, not due to lower cortical bone density. PMID- 16356434 TI - The influence of pulmonary function testing on the management of asthma in children. AB - OBJECTIVE: To assess how often in a single encounter that pulmonary function tests (PFTs) influenced management decisions in children with asthma, beyond what was obtained from history and physical examination alone. STUDY DESIGN: Children with asthma (n = 367, age 4 to 18 years) performed spirometry before clinical evaluation. Physicians and nurse practitioners in the outpatient pulmonary office evaluated the children and made initial treatment recommendations before reviewing the spirometry results. Any changes based on the test results were documented. RESULTS: Spirometry was abnormal in 45% of the visits, related to underlying asthma severity but not to clinical findings. PFT results changed management decisions in 15% of visits. This frequency was not affected by the patient's age, disease severity, symptom control, or exam findings. When spirometry did not change treatment decisions, the provider was more likely to maintain therapy (58%) than to increase (17%) or decrease (24%) therapy. In contrast, when spirometry did change treatment decisions, the provider was more likely to increase therapy (75%) than to maintain (20%) or decrease (5%) therapy. CONCLUSION: Without PFTs, providers often overestimated the degree of asthma control. This incorrect assessment could have resulted in suboptimal therapy. PMID- 16356435 TI - School absenteeism in children with asthma in a Los Angeles inner city school. AB - OBJECTIVE: To investigate the pattern of school absenteeism in asthmatic children within a Los Angeles inner city school. STUDY DESIGN: Five hundred twenty-eight students of predominant Hispanic ethnicity, from a Los Angeles inner city school were divided into 3 groups: known asthma, high probability of asthma, and low probability of asthma using a previously validated instrument. Attendance records of these students were analyzed to determine total and respiratory absences over a year. School records were compared to the corresponding answers on 513 surveys to determine the accuracy of parental responses in regard to their children's absenteeism. RESULTS: Children with known asthma missed on average 2 more days of school than children with low probability of asthma and high probability of asthma. This was only significant in the younger age groups. Survey responses were found to have a 45.6% agreement with school attendance records. Underestimation occurred more often when school-recorded absentee rates were highest. Overestimation occurred more by parents of children with known asthma or a high probability of asthma. CONCLUSION: In a Los Angeles inner city population, younger children with known asthma miss more days of school than those with no asthma. Survey-reported absenteeism is less accurate than school attendance records. PMID- 16356436 TI - Cost of influenza hospitalization at a tertiary care children's hospital and its impact on the cost-benefit analysis of the recommendation for universal influenza immunization in children age 6 to 23 months. AB - OBJECTIVE: To calculate the costs of influenza hospitalization at a tertiary care children's hospital as the basis of a cost-benefit analysis of the new influenza vaccine recommendation for children age 6 to 23 months. STUDY DESIGN: We reviewed the medical records of all patients admitted to Children's Memorial Hospital (CMH) in 2002 diagnosed with influenza. Total hospital costs were obtained from the Business Development Office. RESULTS: Thirty-five charts were analyzed. Both of the 2 patients requiring mechanical ventilation and 4 of 6 patients admitted to the intensive care unit had high-risk underlying medical conditions. Nine children were age 6 to 23 months; 4 of these 9 had no preexisting medical conditions. Had all 18 high-risk children over age 6 months been protected from influenza, approximately $350,000 in hospital charges could have been saved. CONCLUSIONS: Preventing the additional 4 hospitalizations in the otherwise low risk children age 6 to 23 months for whom vaccine is currently recommended would have cost approximately $281,000 ($46/child) more than the hospital charges saved. When all children age 6 to 23 months are considered, influenza vaccination is less costly than other prophylactic measures. Addition of indirect costs, deaths, outpatient costs, and the cost of secondary cases would favor the cost:benefit ratio for influenza vaccination of all children age 6 to 23 months. PMID- 16356437 TI - Noninvasive therapy with helium-oxygen for severe bronchiolitis. AB - OBJECTIVE: To determine whether noninvasive therapy using a helium-oxygen mixture reduces the use of positive-pressure ventilation in the treatment of respiratory failure caused by severe bronchiolitis. STUDY DESIGN: This was a multicenter, randomized, double-blind, placebo-controlled trial that recruited infants in 4 pediatric intensive care units (PICUs). A total of 39 nonintubated infants with severe bronchiolitis caused by respiratory syncytial virus (RSV) were randomly assigned within 8 hours of PICU admission to receive a helium-oxygen mixture (helium group) or an air-oxygen mixture (control group) through an inflatable head hood. The primary study outcome was the requirement for positive pressure mechanical ventilation. Results were compared using Fisher's exact test. RESULTS: No differences were noted between the control and helium groups with respect to age (1.0 vs 1.1 months), prematurity, or family history of asthma or smoking. Positive pressure ventilation was judged necessary for 4 of the 21 (19.0%) infants in the control group and in 4 of the 18 (22.2%) in the helium group (relative risk = 1.17; 95% confidence interval = 0.34 to 4.01). CONCLUSIONS: This study did not detect any differences between the patients in the helium group and the control group with respect to the rate of positive-pressure ventilation. PMID- 16356438 TI - Deletion allele of angiotensin-converting enzyme is associated with increased risk and severity of bronchopulmonary dysplasia. AB - OBJECTIVE: To explore whether the deletion (D) allele of angiotensin-converting enzyme (ACE) is associated with the risk or severity of bronchopulmonary dysplasia (BPD) among very low birth weight (BW) infants. STUDY DESIGN: Infants with a BW < or = 1250 g were prospectively recruited. The D and I (insertion) alleles of ACE were determined using a polymerase chain reaction followed by restriction fragment length polymorphism analysis. RESULTS: Infants with DD/DI genotype of ACE had a (mean +/- SD) birth weight (938 +/- 204 g vs 925 +/- 196 g) and gestational age (28 +/- 3 weeks vs 28 +/- 2 weeks), similar to infants with II genotype of ACE (P > .05). Infants with DD/DI genotype of ACE were more likely to have BPD than infants with II genotype (47% vs 22%, P = .025). Among infants with BPD, ACE DD/DI genotype was more common among infants with moderate or severe BPD compared with infants with mild BPD (74% vs 26%, P = .012). The number of D alleles of ACE correlated directly and positively with the severity of BPD (R = 0.23, P = .045). CONCLUSION: The D allele of ACE is associated with an increased risk and severity of BPD among preterm infants. PMID- 16356439 TI - Sleep-disordered breathing in Prader-Willi syndrome and its association with neurobehavioral abnormalities. AB - OBJECTIVES: To determine the prevalence and type of sleep-disordered breathing among patients with Prader-Willi syndrome (PWS) and its relationship to such neurobehavioral abnormalities as mental retardation, obsessive-compulsive behavior, and conduct disorders. STUDY DESIGN: Polysomnography (PSG) studies were conducted in 13 unselected subjects with PWS (age 1.5 to 28 years). PSG results were compared with tests of behavior and cognition (Development Behavior Checklist [DBC], Auditory Continuous Performance Test [ACPT], and Wechsler Intelligence Scale appropriate for age). RESULTS: Nine of 13 (69%) subjects had > 10 apneas and hypopneas per hour of sleep. Apart from a 2-year-old subject with normal body weight who demonstrated severe central hypopnea in rapid eye movement sleep, the sleep-breathing disturbance was due to upper airway obstruction. Age adjusted body mass index was associated with more severe hypoxemia during sleep (min SaO2, r = -.87, P < .005) and more sleep disruption (arousals/hour of sleep, r = .62, P < .05; sleep efficiency, r = -.66, P < .05). Increasing severity of obstructive sleep apnea (OSA) or sleep disturbance was associated with daytime inactivity/sleepiness and autistic-relating behavior (DBC) and with impulsiveness (ACPT). Unexpectedly, sleep hypoxemia appeared to be predictive of increased performance IQ. CONCLUSIONS: OSA is prevalent among subjects with PWS and is associated with increased body mass, daytime inactivity/ sleepiness, and some behavioral disturbances. PMID- 16356441 TI - Sex differences in the association between sleep and body mass index in adolescents. AB - OBJECTIVE: To determine whether an association between short sleep duration and increased body mass index (BMI) exists in a sample of U.S. adolescents. STUDY DESIGN: Public-use dataset of the National Longitudinal Study of Adolescent Health. Final sample included 4486 adolescents (51% female). Main outcome was BMI transformed into z-scores for age and sex using reference values from the Centers for Disease Control/National Center for Health Statistics. Overweight was defined as > or =95th percentile. Linear and logistic regression models were calculated. Sleep duration was self-reported in hours. A quadratic term for sleep was added to test curvilinear association. Covariates included age, race, parental education, activity and inactivity scores. RESULTS: Among males, linear regression indicated that sleep duration significantly predicted BMI z-score (Beta = -0.08, 95% CI: -0.12, -0.03). Logistic regression indicated that sleep duration predicted risk of overweight among males (OR = 0.90, 95% CI: 0.82, 1.00). Sleep duration was not a significant predictor among females in either regression model. Quadratic term for sleep was not significant for either sex. CONCLUSIONS: Longer sleep duration was weakly associated with lower BMI and risk of overweight among male adolescents only. This sex-related difference may be due to differences in the physiology of puberty or in sleep characteristics. PMID- 16356442 TI - Decreased serum adiponectin: an early event in pediatric nonalcoholic fatty liver disease. AB - OBJECTIVE: To evaluate the relative concentrations of cytokines in pediatric nonalcoholic fatty liver disease (NAFLD). STUDY DESIGN: Thirty children were evaluated at a fasting morning visit to a pediatric research unit. RESULTS: Compared with normal-weight children (n = 12) and children who were overweight (n = 11), children who had presumed NAFLD (elevated Alanine aminotransferase [ALT] with negative work-up) (n = 7) had significantly lower mean serum adiponectin levels (P = .004). Adiponectin negatively correlated with body mass index (r = 0.60, P = .001), insulin (r = -0.74, P < .001), glucose (r = -0.52, P = .004), and ALT (r = -0.53, P = .003). There was no difference between normal-weight, obese, and presumed NAFLD subjects in mean serum tumor necrosis factor alpha and interleukin-6 and -8 concentrations nor in tumor necrosis factor alpha and interleukin-8 and -10 levels in an ex vivo lipopolysaccharide-stimulated system. CONCLUSIONS: Serum adiponectin is reduced in children with elevated ALT, similar to adults. However, children with presumed NAFLD do not have elevated pro inflammatory cytokine levels. This suggests that depressed adiponectin plays a more proximal role than elevated levels of circulating pro-inflammatory cytokines in the development of NAFLD in children. PMID- 16356443 TI - Underdiagnosis of pediatric obesity and underscreening for fatty liver disease and metabolic syndrome by pediatricians and pediatric subspecialists. AB - OBJECTIVES: To evaluate how often general pediatricians, pediatric endocrinologists, and gastroenterologists diagnose children as overweight and how often interventions are provided, including nutritional counseling and screening for nonalcoholic fatty liver disease (NAFLD) and metabolic syndrome. STUDY DESIGN: The study was a retrospective chart review of outpatient visits at 2 academic hospitals. RESULTS: A total of 2256 patient visits were analyzed, including 715 visits by overweight children. Of those 715 visits, 31% resulted in a diagnosis of overweight. Diagnosis of overweight and nutritional counseling were least likely to occur during gastroenterology visits (22% and 13%, respectively, P < .01). Screening for metabolic syndrome was most likely to occur during endocrinology visits (34%; P < .01). Screening for NAFLD was most likely to occur during gastroenterology visits (23%; P < .01). Children age < 5 years and those with a body mass index percentile (BMI%) of 85% to 94% were least likely to receive diagnosis and intervention for overweight. CONCLUSIONS: The majority of overweight children were not diagnosed and did not receive relevant and recommended evaluations and interventions. Specific attention should be focused on providing diagnosis and interventions for overweight children, especially those age < 5 years and with a BMI% of 85% to 94%. PMID- 16356444 TI - Catch-up growth: testing the hypothesis of delayed growth plate senescence in humans. PMID- 16356445 TI - Congenital disorder of glycosylation (CDG)-Ih patient with a severe hepato intestinal phenotype and evolving central nervous system pathology. AB - We present the clinical, molecular, and biochemical diagnosis of a patient with congenital disorder of glycosylation (CDG)-Ih. We report significant brain dysfunction in this multisystem disease, further expanding its complex clinical spectrum. PMID- 16356446 TI - Clinical and biochemical characterization of a patient with congenital disorder of glycosylation (CDG) IIx. AB - We describe a case of congenital disorder of glycosylation with chronic diarrhea, progressive liver cirrhosis, and recurrent infections. Transferrin analysis showed only hyposialylation, but analysis of total serum N-glycans indicated loss of additional sugars, arguing that the latter generates a more informative picture to search for the primary defect. PMID- 16356447 TI - Reduced docosahexaenoic acid synthesis may contribute to growth restriction in infants born to mothers who smoke. AB - In newborn infants, progressive decrease in anthropometric values and impairment of the docosahexaenoic acid (DHA) status was associated with maternal smoking during pregnancy, with a parallel increase of the metabolic precursor of DHA, alpha-linolenic acid. Maternal smoking may impair DHA synthesis which may contribute to reduced fetal growth. PMID- 16356448 TI - Spontaneous liver hemorrhage during laparotomy for necrotizing enterocolitis: a potential role for recombinant factor VIIa. AB - Necrotizing enterocolitis remains a serious condition in very low birth weight infants, particularly in those infants who require surgery. Perioperative hemorrhage is a potentially fatal complication in this population. We describe our experience in 4 premature infants with necrotizing enterocolitis who received recombinant factor VIIa to manage life-threatening intraoperative hemorrhage. PMID- 16356449 TI - Male/female differences in indomethacin effects in preterm infants. AB - To test whether indomethacin prophylaxis has sex-mediated effects on severe intraventricular hemorrhage (grade III and IV) and on long-term outcomes in extremely-low-birth-weight infants. A secondary analysis was performed in the entire "Trial of Indomethacin Prophylaxis in Preterms study" cohort. The results suggest a weak differential treatment effect of indomethacin by sex. PMID- 16356450 TI - Efficacy of bystander cardiopulmonary resuscitation and out-of-hospital automated external defibrillation as life-saving therapy in commotio cordis. AB - We report a child who sustained commotio cordis after being struck by a baseball, and offer documentation of the advantages of having readily available access to bystander cardiopulmonary resuscitation (CPR) and an automated external defibrillator (AED). We suggest that communities and school districts reexamine the need for accessible AEDs and CPR-trained coaches at organized sporting events for children. PMID- 16356451 TI - Pulmonary hypertension of the neonate resistant to inhaled nitric oxide. PMID- 16356452 TI - Severe ocular trauma in a young girl with a paintball pellet. PMID- 16356454 TI - Pediatric hypereosinophilic syndrome (HES) clinically differs from adult HES, but there is a lack of confirmatory laboratory data. PMID- 16356455 TI - Pancreatic enzyme therapy and gastrointestinal symptoms in patients with cystic fibrosis. PMID- 16356456 TI - Safety and accessibility effects of code modifications and traffic calming of an arterial road. AB - The European Road E12 through the community center of Storuman, Sweden was reconstructed in 1999 and 2000. Pedestrian walkways, traffic islands, chicanes of a type referred to as "Danish buns", a roundabout and a two-directional cycle track along the E12 were installed. The purpose of the reconstruction was to improve safety for pedestrians and bicyclists, primarily for children, the elderly and the disabled, and to reduce the barrier effect of the E12 thoroughfare. In May 2000, the code governing the conduct of drivers at marked crosswalks in Sweden became stricter to improve safety and mobility for pedestrians. The combined effect of reconstructions and change of code was analyzed. Traffic behavior was studied at the intersection where the roundabout was constructed. Yield behavior towards pedestrians changed significantly. The difference was even greater with respect to yielding to child bicyclists - from 6% before to 84% after - even though the code change only related to pedestrians. Crash data analysis suggests a minor increase in fall injuries after reconstructions and change of code. Measures of speed, behavioral studies, questionnaires, face-to-face interviews and crash data analysis suggest that safety has increased not only along the E12 but also along adjacent roads. The final conclusion is that a bypass is not needed in a case like Storuman. Traffic calming of the main arterial through the town seems sufficient. PMID- 16356457 TI - Cardiac autonomic modulation in hypertensive patients with Chagas' disease. AB - BACKGROUND: Arterial hypertension and Chagas' disease are prevalent pathologies in Latin America. It has been demonstrated that each one of them may cause cardiac autonomic dysfunction. This study aimed to investigate the pattern of cardiac autonomic modulation in chagasic-hypertensive patients. METHODS: Subjects (n=120) without left ventricular dysfunction were distributed in four groups: healthy control (n=30); hypertensive (n=30); chagasic (n=30) and chagasic hypertensive (n=30). Patients were evaluated by autoregressive spectral analysis of heart rate variability in three different conditions: baseline, cold face and passive tilt tests. Power spectral densities in low (0.04-0.15 Hz) and high (0.15 0.50 Hz) frequency bands were estimated in both absolute and normalized units. RESULTS: Baseline median values (percentile 25 to percentile 75) of mean arterial pressure (in mmHg) were 93.3 (85.0-96.7), 116.7 (*, #) (110.0-129.2), 86.7 (83.3 92.5) and 106.7 (*, #) (106.7-110.0) for healthy control, hypertensive, chagasic and chagasic-hypertensive patients, respectively (*p<0.05 versus healthy control, #p<0.05 against chagasic group). Heart rate at rest did not differ among groups. Regarding to spectral parameters in baseline conditions, the absolute power of high frequency component of heart rate variability of the chagasic-hypertensive group was significantly lower than that found in healthy control and hypertensive patients. There were no differences in spectral parameters responses during cold face test. After passive tilt test, however, decreases in high frequency oscillations and increases in sympathovagal balance (low and high frequency ratio) were significantly lower in hypertensive, chagasic and chagasic hypertensive patients as compared with healthy control. CONCLUSIONS: These data indicate that chagasic-hypertensive patients presented an impairment of cardiac parasympathetic modulation at baseline conditions as well as in response to passive orthostatic stress. PMID- 16356458 TI - Obtaining high sequence coverage in matrix-assisted laser desorption time-of flight mass spectrometry for studies of protein modification: analysis of human serum albumin as a model. AB - Several approaches were explored for obtaining high sequence coverage in protein modification studies performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Human serum albumin (HSA, 66.5kDa) was used as a model protein for this work. Experimental factors considered in this study included the type of matrix used for MALDI-TOF MS, the protein digestion method, and the use of fractionation for peptide digests prior to MALDI-TOF MS analysis. A mixture of alpha-cyano-4-hydroxycinnamic acid and 2,5 dihydroxybenzoic acid was employed as the final matrix for HSA. When used with a tryptic digest, this gave unique information on only half of the peptides in the primary structure of HSA. However, the combined use of three enzyme digests based on trypsin, endoproteinase Lys-C, and endoproteinase Glu-C increased this sequence coverage to 72.8%. The use of a ZipTip column to fractionate peptides in these digests prior to analysis increased the sequence coverage to 97.4%. These conditions made it possible to examine unique peptides from nearly all of the structure of HSA and to identify specific modifications to this protein (e.g., glycation sites). For instance, Lys199 was confirmed as a glycation site on normal HSA, whereas Lys536 and Lys389 were identified as additional modification sites on minimally glycated HSA. PMID- 16356459 TI - Comparative analysis of various in vitro COT kinase assay formats and their applications in inhibitor identification and characterization. AB - Cancer osaka thyroid (COT) is a member of the mitogen-activated protein kinase kinase kinase family of enzymes and plays a pivotal role in tumor necrosis factor alpha production in macrophages. Consequently, COT is considered to be a promising target for antiinflammatory drug discovery. We describe here the development of in vitro COT assays in several formats and the advantages and disadvantages of each. A cascade assay requires very small amounts of enzyme and can provide a useful tool for high-throughput screening, but it is not desirable for compound mechanistic studies due to complicated kinetics. Direct assays are superior to cascade assays and are suitable for both compound screening and mechanistic studies. Among the direct assays, the homogeneous time-resolved fluorescence (HTRF) format is preferred over the radiometric format due to the robustness, throughput, and ease of use of the HTRF format. When the physiological protein substrate MEK1 (MAP/Erk kinase 1) was used to determine inhibitor potencies, false positives were observed due to compound interference by binding to MEK1. Using a MEK1 peptide substrate, these false positives were eliminated. In addition, we describe a simple method to study the ATP competitiveness of compounds. The knowledge gained through our studies with COT, and the methods described for our assays and compound mechanistic studies, can be readily applied to other kinase targets. PMID- 16356460 TI - Unusual chromatographic behavior of oligonucleotide sequence isomers on two different anion exchange HPLC columns. AB - The retention behavior of the unmodified phosphodiester oligonucleotide sequence isomers was investigated on two different anion exchange columns: Biospher GMB 1000Q (based on DEAE-modified glycidyl methacrylate) and PolyWAX LP (based on silica with a crosslinked coating of linear polyethyleneimine). There was a notable difference in retention of oligonucleotides of the same composition but differing in the position of a single base. The most pronounced difference was observed between the oligonucleotides with the variable base in the end and in the center of the sequence. The use of either acetonitrile or 2-propanol as a mobile phase organic modifier did not markedly affect the retention time patterns. Prediction of the retention times of oligonucleotides must take into account the base position as well as identity. This is the first report of such a "same composition different sequence" effect, described for the short peptides, for synthetic oligonucleotides. PMID- 16356461 TI - Kinetic studies of protein-surface interactions: A two-stage model of surface induced protein transitions in adsorbed biofilms. AB - The irreversible adsorption of proteins on artificial surfaces plays an important role in a wide variety of practical problems. The simple analytical models based on definite concepts regarding the mechanisms of interfacial evolution can be used efficiently for characterization of protein-surface interactions by analyzing the intrinsic kinetics of the process. In this article, analytical expressions are derived for the adsorption kinetics that take into account the presence of more than one adsorbed state for proteins in biofilms. It is shown that the experimentally observed dependence of the adsorbed mass on the concentration of protein in solution can be reproduced with this model, and the approach provides a rapid method for obtaining quantitative parameters for the adsorption process. It is shown by analytical approximation of the kinetic curves for fibrinogen adsorption onto an unmodified gold surface studied by a surface plasmon resonance biosensor that this model is in good quantitative agreement with experiments. It is found that the rate of adsorption, controlled mainly by the mass flow from the solution, determines the contribution both to self assembling and spreading, resulting in variations of adsorbed fibrinogen interfacial structures. PMID- 16356463 TI - Adapter annealing to engineer restriction enzyme sites at cloning junctions. PMID- 16356462 TI - Small ubiquitin-like modifying protein isopeptidase assay based on poliovirus RNA polymerase activity. AB - The ubiquitin-proteasome pathway is the major nonlysosomal proteolytic system in eukaryotic cells responsible for regulating the level of many key regulatory molecules within the cells. Modification of cellular proteins by ubiquitin and ubiquitin-like proteins, such as small ubiquitin-like modifying protein (SUMO), plays an essential role in a number of biological schemes, and ubiquitin pathway enzymes have become important therapeutic targets. Ubiquitination is a dynamic reversible process; a multitude of ubiquitin ligases and deubiquitinases (DUBs) are responsible for the wide-ranging influence of this pathway as well as its selectivity. The DUB enzymes serve to maintain adequate pools of free ubiquitin and regulate the ubiquitination status of cellular proteins. Using SUMO fusions, a novel assay system, based on poliovirus RNA-dependent RNA polymerase activity, is described here. The method simplifies the isopeptidase assay and facilitates high-throughput analysis of these enzymes. The principle of the assay is the dependence of the viral polymerase on a free N terminus for activity; accordingly, the polymerase is inactive when fused at its N terminus to SUMO or any other ubiquitin-like protein. The assay is sensitive, reproducible, and adaptable to a high-throughput format for use in screens for inhibitors/activators of clinically relevant SUMO proteases and deubiquitinases. PMID- 16356464 TI - Locked nucleic acids for optimizing displacement probes for quantitative real time PCR. AB - Displacement probes have recently been described as a novel probe-based detection system for use in both quantitative real-time polymerase chain reaction (PCR) and single nucleotide polymorphism genotyping analysis. Previous reports have shown that shorter probes (23 mer) had improved detection sensitivity relative to longer probes (29 mer), with the likely reason for this effect being the improved hybridization kinetics of shorter probes. Sterically modified locked nucleic acids (LNAs) have been used to improve the design of a range of real-time PCR probes by raising the melting temperature (Tm) of the probe and enabling shorter probe designs to be considered. A displacement probe for gapdh was designed and tested successfully, and this probe was then redesigned with LNAs to an 11 mer probe. This probe showed increased detection sensitivity compared with the original 26 mer probe. To detect the widest range of displacement probe designs at maximum sensitivity, we have also developed a novel fluorescence capture two step PCR protocol. This method produces enhanced probe quenching with a single standardized protocol ideal for high-throughput applications. The displacement probes tested produced sensitive and efficient quantitative analyses of template serial dilutions when compared with a range of commercially available predesigned real-time PCR detection systems, including TaqMan MGB probes, QuantiTect MGB probes, and LUX primers. PMID- 16356465 TI - Extracellular metabolomics: a metabolic footprinting approach to assess fiber degradation in complex media. AB - This work reports the implementation and optimization of a method for high throughput analysis of metabolites produced by the breakdown of natural polysaccharides by microorganisms. Our simple protocol enables simultaneous separation and quantification of more than 40 different sugars and sugar derivatives, in addition to several organic acids in complex media, using 50-mul samples and a standard gas chromatography-mass spectrometry platform that was fully optimized for this purpose. As an implementation proof-of-concept, we assayed extracellular metabolite levels of three bacterial strains cultivated on complex medium rich in polysaccharides and under identical growth conditions. We demonstrate that the metabolic footprinting profile data distinguish among sample types such as typical metabolomics data. Moreover, we demonstrate that the differential metabolite-level data provide insight on specific fibrolytic activity of the different microbial strains and lay the groundwork for integrated proteome-metabolome studies of fiber-degrading microorganisms. PMID- 16356466 TI - Calibrating Borg scale ratings of hand force exertion. AB - A study was conducted to assess the efficacy of calibrating subjective worker ratings of hand exertions to reduce error in estimates of applied force. Twenty volunteer subjects applied pinch and power grip forces corresponding to their perceptions of different Borg CR-10 scale levels using both "grip-to-scale" and "guided-grip" procedures. These data were used separately to define relationships between scale ratings and actual force application. Two gripping tasks were performed and corresponding subjective hand force ratings were calibrated using the grip-to-scale calibration data. Results showed that the mean estimation error for a 44.5 N (10 lb) power grip task was significantly reduced from 142.8 (+/ 69.0) to 62.3 (+/-58.3) N. The guided-grip calibration method also significantly reduced rating error for the power grip task, however the estimates were biased toward zero. Neither calibration procedure improved rating accuracy of an 8.9 N (2 lb) pinch grip task. The study results indicate that calibration of hand force ratings using the grip-to-scale procedure may improve the accuracy of hand exertion measurements using the Borg CR-10 scale. PMID- 16356467 TI - Effects of carried weight on random motion and traditional measures of postural sway. AB - The purpose of the study was to investigate the effects of load weight carried by soldiers upon postural sway. Fourteen US Army enlisted men participated. Postural sway and muscle activity were measured while participants stood on a force plate. The load weight conditions, comprised of Army clothing and load-carriage equipment were 6, 16, and 40 kg. With an increase in load weight, stabilogram diffusion analysis revealed that random movement of postural sway decreased. Also, with an increase in load weight, center of pressure excursions increased linearly but muscle activity changed minimally. In short, increasing load weight challenged the load carriers' stability, reduced the randomness of postural sway and required the load carriers to exert greater control of the load in order to maintain balance. PMID- 16356468 TI - Identification and purification of a DNA-binding protein interacting with the promoter of 5'-nucleotidase in Dictyostelium discoideum. AB - The developmental management of 5'-nucleotidase (5nt) expression in Dictyostelium discoideum has provided a focal point for studies of gene regulation at the level of transcription. To identify DNA-protein interactions involved in the 5nt regulation, EMSAs were performed using short oligonucleotides, designed to span a 357bp promoter region. A binding activity (R(f)=0.33) was identified and shown to be specific to the nucleotide sequence between -338 and -309bp relative to 5nt ATG. Characterization of the binding activity, including the effects of salt and temperature, provided insight into the nature and stability of the protein. The protein was purified in a series of chromatographic stages, including DEAE Sephacel, heparin-Sepharose, DNA affinity, and gel filtration. SDS-PAGE analysis identified a polypeptide with a molecular weight of 70kDa. Mass spectrometry revealed that the purified protein was a putative formyltetrahydrofolate synthase. PMID- 16356469 TI - Asn183 in alpha5 is essential for oligomerisation and toxicity of the Bacillus thuringiensis Cry4Ba toxin. AB - The proposed toxicity mechanism of the Bacillus thuringiensis Cry insecticidal proteins involves membrane penetration and lytic pore formation of the alpha4 alpha5 hairpins in the target larval midgut cell membranes. In this study, alanine substitutions of selected polar residues (Tyr(178), Gln(180), Asn(183), Asn(185), and Asn(195)) in the hydrophobic helix-alpha5 of the Cry4Ba mosquito larvicidal protein were initially conducted via PCR-based directed mutagenesis. Upon IPTG induction, all the 130-kDa mutant protoxins were highly expressed in Escherichia coli as cytoplasmic inclusions, with yields similar to the wild-type protoxin. When E. coli cells expressing each mutant toxin were tested against Stegomyia aegypti mosquito larvae, the larvicidal activity of the N183A mutant was almost completely abolished whereas the four other mutants showed only a small reduction in toxicity. Additionally, replacements of this critical residue with various amino acids revealed that the uncharged polar residue at position 183 in alpha5 is crucial for larvicidal activity. Further characterisation of the N183K bio-inactive mutant revealed that the 65-kDa activated toxin was unable to form oligomers in lipid vesicles and its ability to induce the release of entrapped calcein from liposomes was much weaker than that of the wild-type toxin. These results suggest that the highly conserved Asn(183) located in the middle of the transmembrane alpha5 of Cry4Ba plays a crucial role in toxicity and toxin oligomerisation in the lipid membranes. PMID- 16356470 TI - Effects of local injections of vasoactive drugs on eruption rate of incisor teeth in anaesthetized rats. AB - OBJECTIVES: Previous studies have suggested that the vasculature in the dental pulp and periodontal tissues plays an important role in producing the eruptive force in continuously erupting incisors. The aim of the present study was to investigate the effects of local injections of vasoactive drugs on regional blood flow within the socket in association with axial tooth movements to clarify the role of the local vascular system on tooth eruption. DESIGN: Twenty-two male Wistar rats, weighing 309+/-21 g (S.D.), were immobilized with halothane anaesthesia. We measured the regional blood flow within the socket using a laser Doppler flowmeter, and the axial movements of the mandibular incisor using a displacement detector. The local injections of the vasoactive drugs, adrenaline (0.001, 0.01, and 0.1 microg/kg body weight) and acetylcholine (0.05, 0.5, and 5 microg/kg), into the base of the incisor were performed by a microinjector at a rate of 1 microl/kg body weight. RESULTS: The injections of various doses of adrenaline decreased the mean regional blood flow and eruption rate dose dependently, while those of acetylcholine increased the mean regional blood flow and eruption rate dose-dependently. The changes in the regional blood flow and eruption rate were transient. Significant correlations (p<0.001) were obtained between the maximum and minimum values in the regional blood flow and in the eruption rate following injections of various doses of adrenaline and acetylcholine. CONCLUSION: These results support the hypothesis that the eruptive force of the rat incisor is closely related to the vasculature within the socket. PMID- 16356471 TI - Stabilization of trypsin by association to plasma membranes: implications for tryptic cleavage of membrane-bound Na,K-ATPase. AB - Tryptic cleavage has been a potential method for studying the structure and mechanism of many membrane transport proteins. Here, we report tight association of trypsin to pig kidney plasma membranes enriched in Na,K-ATPase. Trypsin also associated with protein-free vesicles prepared from plasma membrane lipids. Membrane-associated trypsin was found to be highly resistant to autolysis and insensitive to inhibition by PMSF. Na,K-ATPase substrate ions differentially influenced the level of trypsin membrane association. Thus, NaCl significantly increased trypsin membrane association compared to KCl. The ions seem to exert direct effects on the membrane independent of their effects on protein conformation. Bicarbonate anions, which detach peripheral membrane proteins, efficiently released trypsin from the membrane. Trypsin membrane association was found to enhance the cleavage of the Na,K-ATPase gamma-subunit. Comparison between membranes from shark rectal gland and pig kidney showed that trypsin association was significantly higher in the former. This was found to be partly due to the presence of higher cholesterol levels in the membrane. In conclusion, the differential membrane association of trypsin may affect the outcome of proteolytic cleavage of membrane-bound proteins. PMID- 16356472 TI - Reduced specificity of autobiographical memories following a negative mood induction. AB - Reduced autobiographical memory specificity (AMS) to emotional and neutral cue words appears to be a stable cognitive marker of clinical depression. For example, reduced AMS is present in remitted/recovered depressed patients and shows no reliable relationship with current levels of depressed mood in correlational studies. The present study examined whether reduced AMS could be induced in healthy volunteers with no history of depression, using a negative mood manipulation and whether levels of AMS and induced mood were positively correlated. Results showed a reduction in AMS following negative mood induction, compared to a neutral induction, whereas positive mood induction had no effects on AMS. Furthermore, lower happiness following the induction phase correlated positively with reduced AMS, and the extent of happiness reduction from pre- to post-induction correlated positively with reduction in AMS. These results suggest that AMS is, at least in part, a function of current emotion state. The implications for the literature on AMS as a stable marker of clinical depression are discussed. PMID- 16356473 TI - Loop model: mechanism to explain partial gene duplications in segmented dsRNA viruses. AB - Gene rearrangements in a head-to-tail fashion have been described several times for gene segments of the rota-, phytoreo-, and orbiviruses. Several mechanisms have been proposed to explain the occurrence of partial duplications, however, none of these models has been fully satisfactory to explain the occurrence of all the observed duplicated genes. Based on recently available structural data about the lambda3 RNA-dependent-RNA-polymerase of reoviruses, we propose the 'loop model' as a plausible explanation for the occurrence of partial gene duplications in dsRNA viruses. PMID- 16356474 TI - Free fatty acids administered into the colon promote the secretion of glucagon like peptide-1 and insulin. AB - We examined whether free fatty acids (FFAs) promote glucagon-like peptide-1 (GLP 1) secretion when administered into the intestinal tract. We found that an unsaturated long-chain FFA, alpha-linolenic acid (alpha-LA), resulted in increased plasma GLP-1 and insulin levels when administered into the colon. Such stimulatory effects were not apparent with either vehicle or a saturated middle chain FFA, octanoic acid (OA). Concomitant with GLP-1 secretion, the administration of alpha-LA, but not vehicle or OA, also resulted in a significant increase in the population of pERK positive cells within the GLP-1 positive cells of the colonic mucosa. Moreover, colonic administration of alpha-LA into normal C3H/He mice caused a reduction in plasma glucose levels, as well as in type 2 diabetic model NSY mice. Our results indicate that the in vivo colonic administration of alpha-LA promotes secretion of incretin GLP-1 by activating the ERK pathway in L-cells and thereby enhances the secretion of insulin. PMID- 16356475 TI - A novel phenotype of eight spores asci in deletants of the prion-like Rnq1p in Saccharomyces cerevisiae. AB - We report that a null rnq1 mutation in the yeast RNQ1 (YCL028w) prion-like gene of so far unknown function produces the doubling of spores in the asci. This phenotype is possibly due to the lack of inhibition by Rnq1p of an additional mitotic division during ascus formation. This novel phenotype termed "octopus asci" could be similar to prion [PIN+] phenotype. PMID- 16356476 TI - Differential gene expression profiling between wild-type and ALAS2-null erythroblasts: identification of novel heme-regulated genes. AB - To identify erythroid-specific heme-regulated genes, we performed differential expression analysis between wild-type and heme-deficient erythroblasts, which had been prepared from wild-type and erythroid-specific delta-aminolevulinate synthase-null mouse ES cells, respectively. Among 8737 clones on cDNA array, 40 cDNA clones, including 34 unknown ESTs, were first selected by their high expression profiles in wild-type erythroblasts, and evaluated further for their erythroid-lineage specificity, expression in hematopoietic tissues in vivo, and heme-dependent expression, which yielded 11, 4, and 4 genes, respectively. Because of the selection strategy employed, the final 4 were considered as the newly identified erythroid-specific heme-regulated genes. These 4 genes were uncoupling protein 2, nucleolar spindle-associated protein, cellular nucleic acid binding protein, and a novel acetyltransferase-like protein. These findings thus suggest that heme may regulate a wide variety of hitherto unrecognized genes, and further analysis of these genes may clarify their role in erythroid cell differentiation. PMID- 16356477 TI - The candidate tumor suppressor CST6 alters the gene expression profile of human breast carcinoma cells: down-regulation of the potent mitogenic, motogenic, and angiogenic factor autotaxin. AB - We recently coined CST6 as a novel candidate tumor suppressor gene for breast cancer. CST6 indeed is expressed in the normal human breast epithelium, but little or not at all in breast carcinomas and breast cancer cell lines. Moreover, ectopic expression of CST6 in human breast cancer cells suppressed cell proliferation, migration, invasion, and orthotopic tumor growth. To obtain insights into the molecular mechanism by which CST6 exhibits its pleiotropic effects on tumor cells, we compared global gene expression profiles in mock- and CST6-transfected human MDA-MB-435S cells. Out of 12,625 transcript species, 61 showed altered expression. These included genes for extracellular matrix components, cytokines, kinases, and phosphatases, as well as several key transcription factors. TaqMan PCR assays were used to confirm the microarray data for 7 out of 11 genes. One down-regulated gene product, secreted autotaxin/lyso phospholipase D, was of particular interest because its down-regulation by CST6 could explain most of CST6's effect on the breast cancer cells. This study thus provides the first evidence that CST6 plays a role in the modulation of genes, particularly, genes that are highly relevant to breast cancer progression. PMID- 16356478 TI - Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer. AB - The low success rate of animal cloning by somatic cell nuclear transfer (SCNT) is believed to be associated with epigenetic errors including abnormal DNA hypermethylation. Recently, we elucidated by using round spermatids that, after nuclear transfer, treatment of zygotes with trichostatin A (TSA), an inhibitor of histone deacetylase, can remarkably reduce abnormal DNA hypermethylation depending on the origins of transferred nuclei and their genomic regions [S. Kishigami, N. Van Thuan, T. Hikichi, H. Ohta, S. Wakayama. E. Mizutani, T. Wakayama, Epigenetic abnormalities of the mouse paternal zygotic genome associated with microinsemination of round spermatids, Dev. Biol. (2005) in press]. Here, we found that 5-50 nM TSA-treatment for 10 h following oocyte activation resulted in more efficient in vitro development of somatic cloned embryos to the blastocyst stage from 2- to 5-fold depending on the donor cells including tail tip cells, spleen cells, neural stem cells, and cumulus cells. This TSA-treatment also led to more than 5-fold increase in success rate of mouse cloning from cumulus cells without obvious abnormality but failed to improve ES cloning success. Further, we succeeded in establishment of nuclear transfer embryonic stem (NT-ES) cells from TSA-treated cloned blastocyst at a rate three times higher than those from untreated cloned blastocysts. Thus, our data indicate that TSA-treatment after SCNT in mice can dramatically improve the practical application of current cloning techniques. PMID- 16356479 TI - Small interference RNA-mediated knockdown of sperm associated antigen 9 having structural homology with c-Jun N-terminal kinase-interacting protein. AB - Recently, we reported a novel testis-specific sperm associated antigen 9 (SPAG9) protein, a new member of the JNK-interacting protein family, having a functional role in sperm-egg fusion [N. Jagadish, R. Rana, R. Selvi, D. Mishra, M. Garg, S. Yadav, J.C. Herr, K. Okumura, A. Hasegawa, K. Koyama, A. Suri, Biochem. J. 389 (2005) 73-82]. NCBI Blast searches revealed SPAG9 nucleotide sequence similarities with ESTs of various cancerous tissues. In the present study, we compared the efficiency of two independent SPAG9 specific small interfering RNA (siRNA) constructs, BS/U6/spag9 and BS/U6/spag9-I, to ablate the SPAG9 expression in mammalian cells. A positive correlation between the ratio of target gene versus siRNA and the suppression of SPAG9 expression was observed. Further, the cotransfection of BS/U6/spag9 with pcDNA-SPAG9 and pFlag-CMV2-JNK-3 resulted in specific suppression of SPAG9 without affecting JNK-3 expression. The present investigation will eventually extend the application of SPAG9 siRNA in in vivo targeting experiments that aim to define the SPAG9 functional genomics in tumor and reproductive biology. PMID- 16356480 TI - Quantification by real-time PCR of developmental and adult myosin mRNA in rat muscles. AB - A real-time RT-PCR assay using newly designed primers was developed to analyze developmental and adult MHC mRNA expression both in skeletal muscles and single fibers. Only 4 ng of total RNA was necessary for the analysis of the relative mRNA expression of MHC genes. Different validation steps were realized concerning both specificity and sensitivity of each primer set, and linearity and efficiency of each real-time PCR amplification. Then, quantification of MHC mRNA in neonatal and adult muscles as well as in single fibers was done by the deltaC(T) method, with CycA gene as the reference gene. Due to a higher sensitivity than that of a competitive PCR method, we demonstrated that this assay is suitable to study very low level of MHC mRNA expression as developmental MHC in adult muscle and to quantify mRNA from very small samples. PMID- 16356485 TI - Metabolism and transporter-mediated drug-drug interactions of the endothelin-A receptor antagonist CI-1034. AB - CI-1034, an endothelin-A receptor antagonist was being developed for pulmonary hypertension. Drug-drug interaction studies using human hepatic microsomes were conducted to assess CYP1A2, CYP2C9, CYP2C19, CYP3A4 and CYP2D6 inhibition potential; CYP3A4 induction potential was evaluated using primary human hepatocytes. CI-1034 moderately inhibited CYP2C9 (IC(50) 39.6 microM) and CYP3A4 activity (IC(50) 21.6 microM); CYP3A4 inhibition was metabolism-dependent. In human hepatocytes, no increase in CYP3A4 activity was observed in vitro, while mRNA was induced 15-fold, similar to rifampin, indicating that CI-1034 is both an inhibitor and inducer of CYP3A4. A 2-week clinical study was conducted to assess pharmacokinetics, pharmacodynamics and safety. No significant changes were observed in [formula: see text] between days 1 and 14. However, reversible elevations of serum liver enzymes were observed with a 50mg BID dose and the program was terminated. To further understand the interactions of CI-1034 in the liver and possible mechanisms of the observed hepatotoxicity, we evaluated the effect of CI-1034 on bile acid transport and previously reported that CI-1034 inhibited biliary efflux of taurocholate by 60%, in vitro. This indicated that inhibition of major hepatic transporters could be involved in the observed hepatotoxicity. We next evaluated the in vitro inhibition potential of CI-1034 with the major hepatic transporters OATP1B1, OATP1B3, OATP2B1, MDR1, MRP2 and OCT. CI-1034 inhibited OATP1B1 (K(i) 2 microM), OATP1B3 (K(i) 1.8 microM) and OATP2B1 activity (K(i) 3.3 microM) but not OCT, MDR1 or MRP2 mediated transport. Our data indicates that CI-1034 is an inhibitor of major hepatic transporters and inhibition of bile efflux may have contributed to the observed clinical hepatotoxicity. We recommend that in vitro drug-drug interaction panels include inhibition and induction studies with transporters and drug metabolizing enzymes, to more completely assess potential in vivo interactions or toxicity. PMID- 16356486 TI - Identification of HCC-22-5 tumor-associated antigen and antibody response in patients. AB - BACKGROUND: Serological identification of antigens by recombinant expression cloning (SEREX) is a promising method used to analyze tumor-associated antigen (TAA). Nineteen primary HCC-associated antigens have been found from a HCC cDNA library using autogenous serum by the SEREX approach. We searched for HCC associated antigens and applied them to HCC diagnosis. METHODS: Nine of 19 primaries HCC-associated antigens identified by SEREX method were tested their immune response again with distinct allogeneic sera. One of the screened HCC associated antigens, HCC-22-5 was recombined and expressed and made the frequency analysis of its seropositivity in various patients using the methods of Western blot and ELISA. RESULTS: SEREX analysis showed that 9 primary HCC-associated antigens had high-titered IgG antibody in the majority of HCC patients. Western Blot method confirmed that 3/7 HCC patients had antibodies against HCC-22-5, which demonstrated that expressed HCC-22-5 antigen had the character of antigen. Sera samples from 341 patients and 80 normal individuals have been tested for autoantibodies against HCC-22-5 by ELISA method. The results found that 51/128 of HCC, 11/76 of chronic hepatitis, 11/22 of liver cirrhosis and 8/54 of nasopharynx cancer patients had antibodies against HCC-22-5. No antibody response to HCC-22-5 had been found in the sera of 7 lung cancers, 54 gastric-intestine patients and 80 normal individuals. The groups of HCC and liver cirrhosis had higher antibody positive rate than that of other groups (p<0.05). In the HCC sera with alpha fetoprotein (AFP) negative, the positive rate of HCC-22-5 was as high as 78.9%. CONCLUSIONS: HCC-22-5 can be used for HCC serologic screening, especially for the patients with AFP negative. PMID- 16356487 TI - Suboptimal tradeoffs in information seeking. AB - Explicit information-seeking actions are needed to evaluate alternative actions in problem-solving tasks. Information-seeking costs are often traded off against the utility of information. We present three experiments that show how subjects adapt to the cost and information structures of environments in a map-navigation task. We found that subjects often stabilize at suboptimal levels of performance. A Bayesian satisficing model (BSM) is proposed and implemented in the ACT-R architecture to predict information-seeking behavior. The BSM uses a local decision rule and a global Bayesian learning mechanism to decide when to stop seeking information. The model matched the human data well, suggesting that adaptation to cost and information structures can be achieved by a simple local decision rule. The local decision rule, however, often limits exploration of the environment and leads to suboptimal performance. We propose that suboptimal performance is an emergent property of the dynamic interactions between cognition and the environment. PMID- 16356488 TI - Na+/K+ -ATPase regulates tight junction formation and function during mouse preimplantation development. AB - Research applied to the early embryo is required to effectively treat human infertility and to understand the primary mechanisms controlling development to the blastocyst stage. The present study investigated whether the Na(+)/K(+) ATPase regulates tight junction formation and function during blastocyst formation. To investigate this hypothesis, three experimental series were conducted. The first experiments defined the optimal dose and treatment time intervals for ouabain (a potent and specific inhibitor of the Na(+)/K(+)-ATPase) treatment. The results demonstrated that mouse embryos maintained a normal development to the blastocyst stage following a 6-h ouabain treatment. The second experiments investigated the effects of ouabain treatment on the distribution of ZO-1 and occludin (tight junction associated proteins). Ouabain treatment (up to 6 h) or culture in K(+)-free medium (up to 6 h) resulted in the appearance of a discontinuous ZO-1 protein distribution and a loss of occludin immunofluorescence. The third set of experiments examined the influence of ouabain treatment on tight junction function. Ouabain treatment or culture in K(+)-free medium affected tight junction permeability as indicated by an increase in the proportion of treated embryos accumulating both 4 kDa and 40 kDa fluorescein isothiocyanate (FITC)-dextran into their blastocyst cavities. The results indicate that the Na(+)/K(+)-ATPase is a potent regulator of tight junction formation and function during mouse preimplantation development. PMID- 16356489 TI - PEPPER, a novel K-homology domain gene, regulates vegetative and gynoecium development in Arabidopsis. AB - Pistil final morphology relies on floral meristem homeostasis, proper organ specification and regional differentiation. These are developmental processes in which sophisticated signaling networks are being uncovered. However, further elements for fine-tuning adjustment still remain to be disclosed. At the molecular level, posttranscriptional modulators may fit such a profile. In this work, we describe the characterization of PEPPER (PEP), a novel Arabidopsis gene encoding a polypeptide with K-homology (KH) RNA-binding modules, which acts on vegetative growth and pistil development. PEP was initially identified as one of the gene functions affected in a complex mutant carrying a chromosomal reorganization, which exhibits aberrant phyllotaxy and small fruits with supernumerary carpels. In contrast, plants carrying single-gene pep null mutations exhibit subtle morphological alterations. Individuals bearing a stronger-than-null allele present a phenotype comprising leaf alterations, phyllotactic errors and sporadic presence of fruits with multiple valves. Accordingly, dynamic PEP expression was detected in all major organs examined. Complementation experiments with a PEP genomic clone confirmed a role for PEP as a regulator in vegetative and reproductive development. Moreover, our genetic studies suggest that PEP interacts with element(s) of the CLAVATA signaling pathway. PMID- 16356490 TI - Scratching behavior in mice induced by the proteinase-activated receptor-2 agonist, SLIGRL-NH2. AB - We examined whether the proteinase-activated receptor-2 (PAR2) agonist, H-Ser-Leu Ile-Gly-Arg-Leu-NH2 (SLIGRL-NH2), could induce scratching behavior in mice. Intradermal injections of SLIGRL-NH2 (10-50 microg) evoked dose dependent scratching. This behavior peaked near 5 min and returned to preinjection levels within 30 min. Pretreatment of animals with a histamine H1 receptor antagonist, pyrilamine, blocked histamine induced scratching, but it had little effect on SLIGRL scratching. Our study suggests that PAR2 mediates histamine independent itch. PMID- 16356491 TI - Influence of the post-ovariectomy time frame on the experimental anxiety and the behavioural actions of some anxiolytic agents. AB - This study analyzes the long-term effects of ovariectomy on the basal experimental anxiety of rats and the influence of this condition on the anxiolytic properties of diazepam and the 5-HT(1A) receptor agonist 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH-DPAT). Rats ovariectomized 3, 6 and 12 weeks previously, were tested in the burying behaviour paradigm and in an automatic activity counter. The highest values of time spent burying were observed in the 12-week group. In general, the 12-week group was more responsive to diazepam than the 3-week group, while 8-OH-DPAT showed similar effects on time spent burying, independently of how long ago the ovariectomy was done. Detection of anxiogenic like behaviours by the shock-probe burying test in rats after chronic absence of gonadal hormones could help in the understanding of mood changes associated with human menopause. PMID- 16356492 TI - Differential in vitro cellular response induced by exposure to synthetic vitreous fibers (SVFs) and asbestos crocidolite fibers. AB - In this study, we analyzed the effects of synthetic vitreous fibers (SVFs) on a mesothelial (MeT5A) and a fibroblast cell line (NIH3T3), compared to those exerted by crocidolite asbestos fibers. SVFs (glass wool, rock wools) do not induce significant changes in cell mortality, whereas crocidolite asbestos fibers caused a dose-dependent cytotoxicity. We investigated the correlation between the fiber-induced cytotoxicity and the extent and type of interaction of the fibers with the cell surface, and we observed that SVFs, unlike crocidolite asbestos fibers, establish few and weak interactions. Moreover, after internalization, crocidolite asbestos fibers are often found free in the cytoplasm, whereas glass wool fibers are mainly localized inside cytoplasmic vacuoles. After treatments, we also detected signs of oxidative stress, revealed by an increased reactive oxygen species (ROS) production and by an induction of superoxide dismutase (SOD) activity. The lipoperoxidative damage was characterized by a decrease in polyunsaturated fatty acids (PUFA), an increase in the content of thiobarbituric reactive species (TBARS) and a consumption of vitamin E, as a lipophilic antioxidant. Furthermore, we investigated the effect of fiber exposure on cell proliferation. and it was found that, unlike crocidolite asbestos fibers, SVFs did not induce a significant increase in DNA synthesis. PMID- 16356493 TI - Modulation of RIZ gene expression is associated to estradiol control of MCF-7 breast cancer cell proliferation. AB - The retinoblastoma protein-interacting zinc-finger (RIZ) gene, a member of the nuclear protein methyltransferase superfamily, is characterized by the presence of the N-terminal PR domain. The RIZ gene encodes for two proteins, RIZ1 and RIZ2. While RIZ1 contains the PR (PRDI-BF1 and RIZ homologous) domain, RIZ2 lacks it. RIZ gene expression is altered in a variety of human cancers and RIZ1 is now considered to be a candidate tumor suppressor. Estradiol treatment of MCF-7 cells produced a selective decrease of RIZ1 transcript and an increase of total RIZ mRNA. Experiments of chromatin immunoprecipitation indicated that RIZ2 protein expression was controlled by estrogen receptor and RIZ1 had a direct repressor function on c-myc gene expression. To investigate the role of RIZ gene products as regulators of the proliferation/differentiation transition, we analyzed the effects of forced suppression of RIZ1 induced in MCF-7 cells by siRNA of the PR domain-containing form. Silencing of RIZ1 expression stimulated cell proliferation, similar to the effect of estradiol on these cells, associated with a transient increase of c-myc expression. PMID- 16356494 TI - Histone deacetylase inhibitor FR901228 enhances the antitumor effect of telomerase-specific replication-selective adenoviral agent OBP-301 in human lung cancer cells. AB - Replication-competent oncolytic viruses are being developed for human cancer therapy. We previously reported that an attenuated adenovirus OBP-301 (Telomelysin), in which the human telomerase reverse transcriptase promoter element drives expression of E1A and E1B genes linked with an internal ribosome entry site, could replicate in and causes selective lysis of human cancer cells. Infection efficiency in target cancer cells is the most important factor that predicts the antitumor effects of OBP-301. The objectives of this study are to examine the effects of the histone deacetylase inhibitor FR901228 on the level of coxsackie and adenovirus receptor (CAR) expression and OBP-301-mediated oncolysis in human non-small cell lung cancer cell lines. Flow cytometric analysis revealed up-regulated CAR expression in A549 and H460 cells following treatment with 1 ng/ml of FR901228, which was associated with increased infection efficiency as confirmed by replication-deficient beta-galactosidase-expressing adenovirus vector. In contrast, neither CAR expression nor infection efficiency was affected by FR901228 in H1299 cells. To visualize and quantify viral replication in the presence of FR901228, we used OBP-401 (Telomelysin-GFP) that expresses the green fluorescent protein (GFP) reporter gene under the control of the cytomegalovirus promoter in the E3 region. Fluorescence microscopy and flow cytometry showed that FR901228 increased GFP expression in A549 and H460 cells following OBP-401 infection in a dose-dependent manner, but this effect did not occur in H1299 cells. In addition, OBP-301 and FR901228 demonstrated a synergistic antitumor effect in A549 cells in vitro, as confirmed by isobologram analysis. Our data indicate that FR901228 preferentially increases adenovirus infectivity via up regulation of CAR expression, leading to a profound oncolytic effect, which may have a significant impact on the outcome of adenovirus-based oncolytic virotherapy. PMID- 16356496 TI - Trichinella spiralis infection affects p47(phox) protein expression in guinea-pig alveolar macrophages. AB - To establish whether NADPH oxidase activation, responsible for previously demonstrated Trichinella spiralis-induced respiratory burst, results from assembling of membrane and cytosolic NADPH oxidase components and/or increased expression of the oxidase complex proteins, the superoxide anion production and expression of the regulatory p47(phox) subunit were measured in cultured alveolar macrophages obtained during T. spiralis infection of guinea pigs. The results demonstrate for the first time helminth parasite-infection-induced stimulation of NADPH oxidase p47(phox) subunit protein expression, with the effect being decreased by in vivo treatment with cyclosporin A, previously shown to inhibit T. spiralis infection-induced respiratory burst in guinea-pig alveolar macrophages. However, although the expression of the p47(phox) subunit protein remained induced during secondary infection, it was accompanied by superoxide anion production that was significantly suppressed in comparison with that observed during primary infection, suggesting suppressive action of T. spiralis on host's alveolar macrophage immune response, presumably connected with NADPH oxidase complex activity attenuation. PMID- 16356495 TI - Echinostoma caproni: intestinal pathology in the golden hamster, a highly compatible host, and the Wistar rat, a less compatible host. AB - The histopathological changes induced by Echinostoma caproni (Trematoda: Echinostomatidae) in a high (golden hamster) and a low compatible host (rat) were compared at 15 and 30 days post-infection. Infection of rats was characterized by a progressive increase in erosion of villi and elevated numbers of goblet cells, which could be related to the early expulsion of the parasite in a host of low compatibility. In contrast to rats, the number of goblet cell in E. caproni infected hamsters was low, but increased numbers of neutrophils and mesenteric inflammatory cells were observed. This indicated that local inflammatory responses in hamsters were greater than in rats. An immunohistochemical study using polyclonal IgG anti-E. caproni excretory-secretory antigens demonstrated a greater level of passage of E. caproni antigens through the intestinal mucosa in hamsters than in rats, probably in relation to the greater inflammatory response. Our results indicate the fact that early inflammatory responses could be important for the establishment of E. caproni chronic infections in highly compatible hosts. PMID- 16356497 TI - Distribution of IgG subclass antibodies specific for Plasmodium falciparum glutamate-rich-protein molecule in sickle cell trait children with asymptomatic infections. AB - Polymorphism in the beta-globin gene (hemoglobin S) has been associated with protection against severe forms of malaria. In a cross-sectional study, 180 young Gabonese children with and without sickle cell trait and harboring asymptomatic Plasmodium falciparum infections, were assessed for the responses to recombinant protein containing the conserved region of glutamate-rich protein (GLURP). We reported increased age-dependence of antibody prevalence and levels of total IgG (p<0.0001), IgG1 (p=0.009), and IgG3 (p<0.03) antibodies to GLURP with a cut-off at 5 years of age. Whatever the hemoglobin type, cytophilic antibodies (IgG1 and IgG3) were prevalent, but GLURP-specific IgG4 antibodies were detected at significantly (p<0.05) lower levels in HbAS children. We showed that the distribution of non-cytophilic IgG antibodies differs according to the hemoglobin type and to the malaria antigens tested. This may have possible implication for the clearance of malaria parasites and for protection against severe malaria. PMID- 16356498 TI - The dawn and evolution of hormones in the adenohypophysis. AB - The adenohypophysial hormones have been believed to have evolved from several ancestral genes by duplication followed by evolutionary divergence. To understand the origin and evolution of the endocrine systems in vertebrates, we have characterized adenohypophysial hormones in an agnathan, the sea lamprey Petromyzon marinus. In gnathostomes, adrenocorticotropin (ACTH) and melanotropin (MSH) together with beta-endorphins (beta-END) are encoded in a single gene, designated as proopiomelanocortin (POMC), however in sea lamprey, ACTH and MSH are encoded in two distinct genes, proopoicortin (POC) gene and proopiomelanotropin (POM) gene, respectively. The POC and POM genes are expressed specifically in the rostral pars distalis (RPD) and the pars intermedia (PI), respectively. Consequently, the final products from both tissues are the same in all vertebrates, i.e., ACTH from the PD and MSH from the PI. The POMC gene might have been established in the early stages of invertebrate evolution by internal gene duplication of the MSH domains. The ancestral gene might be then inherited in lobe-finned fish and tetrapods, while internal duplication and deletion of MSH domains as well as duplication of whole POMC gene took place in lamprey and gnathostome fish. Sea lamprey growth hormone (GH) is expressed in the cells of the dorsal half of the proximal pars distalis (PPD) and stimulates the expression of an insulin-like growth factor (IGF) gene in the liver as in other vertebrates. Its gene consists of 5 exons and 4 introns spanning 13.6 kb, which is the largest gene among known GH genes. GH appears to be the only member of the GH family in the sea lamprey, which suggests that GH is the ancestral hormone of the GH family that originated first in the molecular evolution of the GH family in vertebrates and later, probably during the early evolution of gnathostomes. The other member of the gene family, PRL and SL, appeared by gene duplication. A beta-chain cDNA belonging to the gonadotropin (GTH) and thyrotropin (TSH) family was cloned. It is expressed in cells of the ventral half of PPD. Since the expression of this gene is stimulated by lamprey gonadotropin-releasing hormone, it was assigned to be a GTHbeta. This GTHbeta is far removed from beta-subunits of LH, FSH, and TSH in an unrooted tree derived from phylogenetic analysis, and takes a position as an out group, suggesting that lampreys have a single GTH gene, which duplicated after the agnathans and prior to the evolution of gnathostomes to give rise to LH and FSH. PMID- 16356499 TI - Polymorphism of the growth hormone gene of red deer (Cervus elaphus). AB - In mammals, pituitary growth hormone (GH) is usually encoded by a single gene, but in some caprine ruminants there are two GH genes, and higher primates have a cluster of at least 5 GH-like genes. We have previously shown that in several artiodactyls (chevrotain, giraffe, and hippopotamus) there are two GH gene sequences, differing by 5-21 nucleotides (nt), but whether these arise from two distinct gene loci is unclear. We report here that in the red deer (Cervus elaphus) also there are two main GH gene sequences (designated A and B) differing at about 23 nt. Investigation of DNA from a number of individual animals demonstrated that this variation was due to allelic polymorphism, with individuals carrying either the A-type or the B-type sequence, or both. A- and B type sequences showed some variation between individuals. The overall difference between the A and B sequences is substantial-greater than that between the GH gene sequences of three distinct bovine species, Bos taurus (ox), Bos indicus (zebu) and Bos grunniens (yak). The biological significance of the presence of two markedly differing GH gene sequences in red deer is not clear, but it is notable that several of the differences between the A and B sequences occur in the 5' upstream region, which may be associated with differences in gene expression. PMID- 16356500 TI - The effect of IV cannula length on the rate of infusion. AB - According to the Hagen-Poiseuille's law, rate of laminar flow through a tubular structure varies directly with fourth power of its radius and inversely with its length. Although it is well recognised that faster infusion rates can be achieved with wider-bore IV cannulae, the effect of length on flow rates is less well known. In the current in vitro study, we assessed the effect of length of an IV cannula on the rate of flow of infusion. Mathematical calculations performed using Hagen-Poiseuille's law predicted an increase of 40% in flow rates when the IV cannulae were shortened by 13mm. However, when the flow rates of the shortened cannulae were measured in vitro an increase of only 4-18% was observed. Although the increase in flow rates was statistically significant, it may not be sufficient to be significant in clinical practice. Turbulence resulting from design characteristics of the infusion system is responsible for the measured flow rates to be lower than that predicted by mathematical calculations. PMID- 16356501 TI - Sexual abuse and reproductive health among unmarried young women seeking abortion in China. AB - OBJECTIVE: To investigate sexual violence and its impact on reproductive health in unmarried young women seeking abortion in China. METHODS: A total of 2002 participants were surveyed by questionnaire, gynecologic examination, and laboratory tests for sexually transmitted disease (STD). RESULTS: Overall, 14% of participants had experienced sexual violence and 43.4% were diagnosed with STD. Among victims of sexual abuse, 8.6% had their first sexual encounter when they were younger than 18 years; 42.7% had had 2 or more sexual partners; and 21.6% never used contraception. Multivariate analysis revealed that sexual abuse, multiple sexual partners, sexual activity before the age of 18 years, and not using contraception were important indicators of the presence of STD. CONCLUSION: The prevalence of sexual abuse is high in China; and among unmarried young women seeking abortion, those who experience sexual abuse are at significantly increased risk for STD. PMID- 16356502 TI - Stage IV cystocele treated by sacrocolpopexy. PMID- 16356503 TI - Tacit knowledge of public health nurses in identifying community health problems and need for new services: a case study. AB - BACKGROUND: The purpose of this study was to explore the tacit knowledge of public health nurses in identifying community health problems and developing relevant new projects. Previous research only roughly showed those skills for creating new community health services, such as lobbying. METHOD: Nine Japanese public health nurses who had created new projects in their municipalities were selected by theoretical sampling and interviewed in 2002-2003. Yin's Case Study Method, especially the multiple-case study design, was used. RESULTS: All 9 public health nurses used similar approaches in identifying community health problems and the need for creating new services, even though their experiences differed and the kinds of projects varied. They identified the difficulties of clients, recognized clients who had the same problems, elucidated the limitations of existing services, and forecasted outcomes from the neglect of the clients' problems. Then they succeeded in creating a new project by examining individual health problems in the context of their community's characteristics, societal factors, and using existing policies to support their clients. CONCLUSION: This is the first study to explore the skills of public health nurses and their intention to use such skills in creating new projects as well as the exact process. They could identify community health problems that will be the basis for developing new services to provide care for individual clients. This is different from the traditional community assessment approach that requires the collection of a huge amount of information to clarify community health problems. The tacit knowledge of public health nurses will help to create needs-oriented new services more smoothly. PMID- 16356504 TI - Examination of genetic effects of polymorphisms in the MCP-1 and CCR2 genes on MI in the Icelandic population. AB - Monocyte chemotactic protein-1 (MCP-1) and its receptor CCR2 may play a role in mediating atherosclerosis. The polymorphisms MCP-1 -2518A>G and CCR2 190G>A have been reported to be associated with increased risk for developing atherosclerosis. The aim of this study was to investigate the effect of these polymorphisms and the MCP-1 -2076A>T polymorphism on the development of myocardial infarction (MI) in an Icelandic cohort. Four hundred and sixty MI survivors and 1842 disease free controls were genotyped. No significant difference in the frequencies of any of the polymorphisms between the cases and the controls was found, with OR=0.87 (95% CI 0.71-1.08) for MCP-1 -2518G, OR=1.05 (95% CI 0.84-1.33) for MCP-1 -2076T and OR=0.93 (95% CI 0.71-1.23) for CCR2 190A. An effect of an OR for MI of 1.4 for the MCP-1 -2518G, MCP-1 -2076T and CCR2 190A alleles could be detected in a cohort of the size of this study with power=0.8 and alpha=0.05. In our cohort we were unable to demonstrate a significant association of the MCP-1 -2518A>G, MCP-1 -2076A>T or CCR2 190G>A polymorphisms with MI. These results do not support the extent of risk associated with developing MI previously reported for the CCR2 190 and MCP-1 -2518 polymorphisms. PMID- 16356505 TI - Chylomicron remnants upregulate CD40 expression via the ERK pathway and a redox sensitive mechanism in THP-1 cells. AB - CD40 is a 48kDa phosphorylated transmembrane glycoprotein that belongs to the tumor necrosis factor receptor superfamily and may play a role in formation of atherosclerotic plaques. Here, we investigated the effect of chylomicron remnants on CD40 expression in the human premonocytic cell line, THP-1 cells. Chylomicron remnants upregulated the expression of CD40 protein and mRNA in a dose- and time dependent manner. Further, chylomicron remnants increased the generation of reactive oxygen species as determined by an increasing level of 2',7' dichlorofluorescein. Pretreatment with the antioxidant, N-acetylcysteine, inhibited chylomicron remnant-induced CD40 protein expression by 60%. On the other hand, chylomicron remnants transiently increased the phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and p38 mitogen-activated protein kinase (MAPK). Pretreatment with the MAPK kinase inhibitor, U0126, completely inhibited chylomicron remnants-induced CD40 protein expression, whereas the p38 MAPK inhibitor, SB203580, had no effect. Pretreatment with N-acetylcysteine had no effect on chylomicron remnant-induced ERK 1/2 phosphorylation. These data suggest that CD40 expression stimulated by chylomicron remnants in THP-1 cells is dependent on ERK 1/2-mediated pathway, which is followed by redox-sensitive mechanism-dependent and independent pathway. Thus, chylomicron remnants may contribute to the formation of atherosclerotic plaques via their immunological and proinflammatory effects. PMID- 16356506 TI - Force plate for measuring the ground reaction forces in small animal locomotion. AB - The importance of kinetic force plate studies of locomotion in small animals has grown recently with the increasing use of rodent models for studies of musculoskeletal diseases. However, the force plates for use with animals much smaller than a cat are difficult to design and use. Here we present data on a commercially available small force plate that accurately collects whole-body and, in a modified form, single-limb ground reaction forces in mice. The method used here is convenient, inexpensive, and readily adaptable for use with a variety of small species. PMID- 16356507 TI - Chromatographic analysis of lignans. AB - Methods and procedures for analysis of lignans in trees and other plants are reviewed. The importance of cautious sample handling and pretreatment procedures to avoid contamination, loss of sample, and unwanted chemical reactions is discussed. Sequential extraction with a non-polar solvent followed by extraction with acetone or ethanol is recommended to separate the lignans from the plant matrix. An additional step of acid, alkaline, or enzymatic hydrolysis may be necessary for some plant matrixes. Flash chromatography is a convenient method for preparative separation and isolation of pure lignans from raw extracts. TLC is very suitable for qualitative screening of extracts and for monitoring of lignan isolation and purification steps. Trimethylsilyl ethers of lignans can be separated and quantified by GC even in the case of complex mixtures of lignans and other polyphenols, and the lignans can be identified by GC-MS in a routine manner. HPLC on reversed-phase columns is especially suited for analysis of lignans and their metabolites in biological matrixes. The recent development of HPLC-electrospray ionisation (ESI)-iontrap MS (MS(n)) and corresponding techniques with high sensitivity and selectivity has proven valuable in lignan analysis. Lignan enantiomers can be separated on chiral HPLC columns. PMID- 16356508 TI - Multiscale-linking simulation of irreversible colloidal deposition in the presence of DLVO interactions. AB - An efficient multiscale-linking algorithm, based on the self-consistent integration of Brownian dynamics simulation of particle trajectories with the solution of the continuum-level conservation equation for particle concentration subject to an adaptive Neumann boundary condition that accounts for the blocking effect of deposition, is developed. The algorithm has been already validated in the case of deposition of noninteracting hard spheres [R.V. Magan, R. Sureshkumar, Multiscale Model. Simul. 2 (2004) 475]. In this study, the above algorithm is extended to incorporate particle interactions modeled by the DLVO theory. The simulations are used to identify a time scale at which the deposition process transitions from a power-law to an asymptotic regime. Detailed characterization of the two regimes is provided for a wide range of ionic strength, particle surface charge density, bulk volume fraction, and substrate potential values. The radial distribution functions obtained for various ionic strengths can be collapsed into a master curve when the radial distance is normalized with respect to a characteristic length scale of inter-particle repulsion. Moreover, simulation results suggest a rescaled, uniformly valid soft random sequential adsorption (RSA) model. Simulation results for the kinetics and monolayers structure compare favorably with experimental data, without the use of adjustable parameters. Comparison with other dynamic simulation techniques shows that while their predictions are qualitatively similar, notable quantitative differences exist especially for small ionic strengths. PMID- 16356509 TI - Investigation on gamma-cyclodextrin nanotube induced by N,N'-diphenylbenzidine molecule. AB - Dynamic light scattering (DLS) measurement provides an effective way to investigate the formation of nanotube of gamma-cyclodextrin (gamma-CD) induced by N,N'-diphenylbenzidine (DPB) in water. With the combination of steady-state fluorescence and fluorescence anisotropy experiments, it was found that for alpha and beta-CD, only 1:2 (guest:host) inclusion complexes were formed and for gamma CD, cyclodextrin nanotube was formed involving 16 gamma-CD units at maximum. The pH effect studies with both DLS and fluorescence anisotropy measurements indicated that the hydrogen bonding between neighboring CDs was necessary to the formation of cyclodextrin nanotube. In the temperature experiment, we found that the nanotube of DPB-gamma-CD could exist stably at relatively high temperatures and the transition point for structural collapse was estimated to be around 54 degrees C. The aggregation states of both gamma-CD itself and DPB-gamma-CD nanotube were observed with TEM. PMID- 16356510 TI - Self-assembly of amino-functionalized monolayers on silicon surfaces and preparation of superhydrophobic surfaces based on alkanoic acid dual layers and surface roughening. AB - Reproducibly smooth amino-functionalized surfaces were obtained by deposition of aminopropyltrimethoxysilane (APTMS) at the vapor/solid interface. Characteristics of these amino-functionalized surfaces were evaluated based on atomic force microscopy, water contact angle measurement and X-ray photoelectron spectroscopy. The results showed that APTMS modified surfaces are very homogeneous and the chemical reactivity of modified surfaces can be ensured with high free amino content. Furthermore, for the purpose of tailoring the wettability of silicon surface, dual self-assembled films were achieved by performing reaction between amino-functionalized surface and n-alkanoic acids with different chain length. The wettability of the self-assembled films can be adjusted with altering the hydrocarbon chain length of alkanoic acids. Moreover, cooperation of dual self assembled films with surface roughening, superhydrophobic surfaces with CA larger than 153 degrees were obtained. Thus, the wettability of modified surfaces can be altered greatly with changing hydrocarbon chain length of self-assembled films. PMID- 16356511 TI - Detection of neutralizing anti-therapeutic protein antibodies in serum or plasma samples containing high levels of the therapeutic protein. AB - Neutralizing antibodies against therapeutic proteins can be potentially harmful if the antibody blocks not only the therapeutic activities of the therapeutic protein but also the normal functions of the endogenous counterpart. Detection of the neutralizing anti-therapeutic protein antibodies generally relies on bioassays measuring changes in the biologic activity of the therapeutic protein triggered by the presence of the antibody. Most of the bioassays, particularly the cell-based in vitro assays, fail to detect neutralizing anti-therapeutic protein antibodies when the remaining therapeutic protein level in the assay samples is high. The remaining therapeutic protein, either a free molecule or an immune complex with anti-therapeutic protein antibodies, can inhibit the neutralizing activity of the antibody and prevent detection. We describe the development of a procedure that uses acid dissociation and affinity adsorption to remove therapeutic protein from assay samples. With this procedure, we can detect the presence of neutralizing anti-therapeutic protein antibodies from samples containing high levels of therapeutic protein. PMID- 16356512 TI - Myosin phosphatase isoform switching in vascular smooth muscle development. AB - We are using the myosin phosphatase targeting subunit (MYPT1) as a model gene to study smooth muscle phenotypic diversity. Myosin phosphatase (MP) is the primary effector of smooth muscle relaxation, and MYPT1 is a key target of signals that regulate smooth muscle tone. In a model of portal hypertension we previously showed dynamic changes in the expression of MYPT1 isoforms in the portal vein and upstream mesenteric artery. We hypothesized that this represents a reversion to the fetal phenotype characteristic of muscle hypertrophy. Here we studied MP during vascular smooth muscle phenotypic specification. Between postnatal days 6 and 12 the expression of MYPT1 increased approximately twofold in portal vein with a similar increase in MP activity. MYPT1 switched from C-terminal leucine zipper (LZ) positive to LZ negative splice variant isoforms. This was concordant with a switch from sensitive (10(-7) M) to resistant to cGMP-mediated vascular relaxation. This is consistent with the model in which the MYPT1 C-terminal LZ is required for cGMP-dependent activation of MP. Concordant changes in the expression of other contractile proteins were consistent with a switch from a slow-tonic to a fast-phasic contractile phenotype. In contrast aortic smooth muscle throughout development expressed the MYPT1 LZ positive isoform and relaxed to cGMP. We propose that MP isoform switching during neonatal vascular smooth muscle phenotypic specification may determine changing vascular responses to NO/cGMP signaling in the transition from the fetal to the adult circulation. PMID- 16356513 TI - Can sodium hypochlorite reduce the risk of species introductions from diapausing invertebrate eggs in non-ballasted ships? AB - Many transoceanic vessels enter the Great Lakes carrying residual ballast water and sediment that harbours live animals and diapausing eggs. In this study, we examine the potential for sodium hypochlorite (NaOCl) to reduce the risk of species introductions from diapausing invertebrate eggs in residual ballast sediment. We collected sediment from three transoceanic vessels and from Lake Erie and exposed them to NaOCl concentrations between 0 and 10,000 mg/L for 24 h. Hatching success was reduced by >89% in all four experiments at 1,000 mg/L relative to unexposed controls. Fewer species hatched at high than at low NaOCl concentrations. Based on an average residual ballast of 46.8 tonnes, the volume of NaOCl required to treat inbound vessels is 374 L. Impacts of NaOCl use could be minimized by neutralization of treated residuals with sodium bisulfite. Further research is needed, however, to evaluate the effect of NaOCl on ballast tank corrosion. PMID- 16356514 TI - Imposex in Hexaplex (Trunculariopsis) trunculus (Gastropoda: Muricidae) from the Ria Formosa lagoon (Algarve coast-southern Portugal). PMID- 16356515 TI - The spatial representation of numerical and non-numerical sequences: evidence from neglect. AB - Psychophysical and neuropsychological studies have revealed that humans represent numbers along a continuous, left-to-right oriented mental line. However, it has been recently claimed that this format of representation is not special to numbers because non-numerical sequences would be spatially coded in the same way. To test this hypothesis, the present study investigated the effects of left neglect upon the bisection of numerical and non-numerical intervals. Eight patients with left neglect performed a visual line bisection task and three mental bisection tasks with number, letter, and month intervals. The error pattern in the number bisection task, indexed by the modulating effect of interval length, mirrored that of the visual task and confirmed the left-to-right spatial orientation of the mental number line. In contrast, the bisection of non numerical intervals showed a very different pattern. The results suggest that the spatial layout characterizing numerical representations constitutes a specific property of numbers rather than a general characteristic of ordered sequences. PMID- 16356516 TI - The orexigenic effect of peripheral ghrelin differs between rats of different age and with different baseline food intake, and it may in part be mediated by the area postrema. AB - Ghrelin is mainly secreted during fasting. While an orexigenic effect of peripherally injected ghrelin has been reported, reproducing this effect has often proven difficult. Here, we hypothesized that ghrelin's effect to increase food intake may depend on the experimental conditions (e.g., age of animals). We therefore investigated the effect of an IP ghrelin injection (100 microg/kg) on food intake in rats of different age and at different times during the light-dark cycle, i.e. with different levels of baseline food intake. Ghrelin injected at dark onset in ad libitum fed young rats (body weight [BW] 92 g) slightly increased feeding while no such effect was observed in 12 h food deprived rats (BW 150 g). In the middle of the light phase, ghrelin significantly increased feeding up to 2 h after injection in ad libitum fed rats (BW 130 g; food intake 1 h after injection: NaCl 0.4 +/- 0.2 g versus ghrelin 1.2 +/- 0.3 g [p < 0.05]). In various subsequent experiments, older rats (BW 300-490 g) tested under the same conditions did not respond to a single ghrelin injection. However repeated ghrelin injection (15 microg/kg/day once daily at light onset) over 10 days significantly increased food intake in rats (BW 400-460 g) starting from day 4 of the experiment (24 h food intake: NaCl approx. 19.5 g, ghrelin 22.5 g). Interestingly, the latter effect was completely abolished in rats lesioned in the area postrema (AP). Cumulative food intake was also increased in SHAM but not in AP-X animals (e.g., after 7 days: SHAM/NaCl 135.1 +/- 5.3 g versus SHAM/ghrelin 149.7 +/- 3.5 g [p < 0.05], AP-X/NaCl 127.2 +/- 16.4 versus AP-X/ghrelin 127.9 +/ 5.3). We conclude that ghrelin's effect to increase food intake can best be demonstrated when basal food intake is low. Ghrelin increases feeding mainly in young, fast growing animals. Ghrelin may therefore link the high energy needs to body growth in young individuals. In older animals, peripheral ghrelin increased feeding when injected repeatedly over several days. At least under these conditions, ghrelin's effect was mediated by the AP/NTS region. Using repeated administration, ghrelin might be an interesting tool to increase feeding in patients suffering from wasting diseases such as cancer anorexia. PMID- 16356517 TI - Secondary metabolites from Ganoderma lucidum and Spongiporus leucomallellus. AB - The hydrodistillates and solvent extracts of the fruit bodies of Ganoderma lucidum (Fr.) P. Karst. and Spongiporus leucomallellus (Murril) A. David were investigated. The constituents in both oils comprised hydrocarbons, monoterpenes, sesquiterpenes, and fatty acids. Major volatiles of G. lucidum were trans anethol, R-(-)-linalool, S-(+)-carvone and alpha-bisabolol, while the essential oil of S. leucomallellus contained relatively large amounts of R-(-)-1-octene-3 ol, R-(-)-linalool, 1-hepten-3-one and (Z)-nerolidol. From the n-hexane extract of G. lucidum, the steroid ester ergosta-7,22-diene-3beta-yl pentadecanoate could be identified. From S. leucomallellus two constituents showing structures of 3,4 seco-lanostane type triterpene acids were identified as (+)-23-oxo-3,4-seco lanosta-4(28),7(8),9(11),24(31)-tetraene-3,26-dicarboxylic acid and (+)-20 hydroxy-23-oxo-3,4-seco-lanosta-4(28),7(8),9(11),24(31)-tetraene3,26-dicarboxylic acid, respectively. Cytotoxicity and antimicrobial activity of selected compounds were investigated using standard tests. PMID- 16356518 TI - Occurrence of Leishmania infantum cutaneous leishmaniasis in central Tunisia. AB - Cutaneous leishmaniasis (CL) due to Leishmania infantum occurs sporadically in Tunisia where its distribution is confined to the northern parts of the country. However, during the past decade there have been occasional repeated reports of cases from areas in central Tunisia, known to be free of CL. Epidemiological, clinical and parasitological data regarding these patients were collected and analysed. Data were very suggestive of the sporadic form of CL due to L. infantum. The parasites contained within the lesions of some of the patients were characterised by two different previously described PCR assays, each having different resolutive powers. The first assay, which amplified complete kDNA minicircles, showed a fragment size characteristic of the L. donovani complex; whilst the second consisted of a PCR-RFLP analysis targeting the gp63 coding sequences that confirmed assignment of the parasites to L. infantum species while illustrating its differences from the reference isolate. These findings confirm the aetiology of CL in the concerned areas in central Tunisia and suggest that L. infantum CL might be more prevalent and widespread than previously thought, or possibly emerging in these areas. PMID- 16356519 TI - Risk factors for infection during a dengue-1 outbreak in Maui, Hawaii, 2001. AB - Autochthonous dengue virus transmission, last identified in the state of Hawaii in 1945, was detected again in 2001. A seroepidemiological survey in a high incidence community (Nahiku) and a nearby low-incidence community (Hana Subdivision) was implemented. The two communities studied differed in median household size (two vs. four persons), median lot size (2.8 vs. 0.8acres), proportion of households with mosquito larvae (81 vs. 28%) and incidence of recent infection (39% [28/72] vs. 1% [1/131]). The average number of reported anti-mosquito actions by residents of both locations remained low, and approximately 50% (42/80) of the inspected houses had larvae, evidencing the need for more effective community mosquito control. Logistic regression analysis of risk factors for infection in Nahiku identified residing in properties with birds in the house or yard as significantly associated with infection (odds ratio 7.0, 95% CI 1.7-28.5), probably as an indicator of unspecified environmental characteristics that were attractive to the vector. We documented that nearly 40% of Nahiku residents had acquired dengue locally in 2001 and that undetected dengue outbreaks had occurred in Hawaii. Our data suggest that ecological characteristics may help Hawaii health officials identify communities at increased risk of dengue infection. PMID- 16356520 TI - KChIP3: a binding protein for Taiwan banded krait beta-bungarotoxin. AB - Using B1 chain of beta-bungarotoxin (beta-Bgt) as bait in yeast two-hybrid screen, we found that KChIP3 was a binding protein of B1 chain. Thus, protein protein interaction between beta-Bgt and KChIP3 is investigated in the present study. Pull-down assay showed that recombinant KChIP3 proteins were associated with beta-Bgt as well as B1 chain, whereas the inability of KChIPs 1, 2 and 4 to bind with beta-Bgt was observed. Although Ca2+ was not a crucial factor essential for the binding of KChIP3 with beta-Bgt and B1 chain, their interaction could be enhanced by the addition of Ca2+. Alternatively, the association of A1 chain of beta-Bgt with KChIP3 was marginally detected. The dissociation constant of beta Bgt with KChIP3 were 12.2 and 6.08 microM in the absence and presence of 2mM Ca2+, respectively. Moreover, native KChIP3 from rat brain was to be isolated by beta-Bgt-Sepharose. These observations indicate that KChIP3 is a binding protein of beta-Bgt. In view of the multiple functions of KChIP3 in neuronal cells, the interaction of KChIP3 with beta-Bgt may represent an event for the manifestation of the biological activities of beta-Bgt. PMID- 16356521 TI - The neutralizing effect of a polyclonal antibody raised against the N-terminal eighteen-aminoacid residues of birtoxin towards the whole venom of Parabuthus transvaalicus. AB - Scorpion venom is composed among other things of a large number of neurotoxic peptides affecting all major types of ion channels. The majority of the toxicity of the venom is attributed to the presence of these peptides. In our previous studies using a combination of HPLC and mass spectrometry, we showed that birtoxin like peptides are the major peptidic components of the venom of Parabuthus transvaalicus. These peptides are quite similar to each other differing by only few amino acid residues. In addition they all share a common N terminus of eighteen amino acid residues. We hypothesize that neutralization of this domain will decrease the toxicity of the whole venom of P. transvaalicus. Polyclonal antibodies against the common N-terminal region of the peptides are generated. Here we show by bioassays that the polyclonal antibodies neutralize the venom of P. transvaalicus in a dose dependent manner and by mass spectrometry and western blotting that these peptides indeed react with the polyclonal antibodies. Previously antibodies generated against a single major toxic component of venom have proven to be an effective strategy for antivenin production. In the case of P. transvaalicus the generated antibody is against the majority of the peptidic fraction due to the presence of several highly similar and highly toxic components in this venom. We show that using the knowledge obtained through biochemical characterization studies it is possible to design very specific antibodies that will be useful for clinical applications against Parabuthus envenomation. PMID- 16356522 TI - First report on cylindrospermopsin producing Aphanizomenon flos-aquae (Cyanobacteria) isolated from two German lakes. AB - Three single-filament isolates of Aphanizomenon flos-aquae from two German lakes were found to produce remarkable amounts of the cyanobacterial hepatotoxin cylindrospermopsin (CYN). CYN-synthesis of the strains were evidenced both by LC MS/MS analysis and detection of PCR products of gene fragments which are implicated in the biosynthesis of the toxin. The strains contain CYN in the range of 2.3-6.6 mg g(-1) of cellular dry weight. To our knowledge this is the first report of CYN in A. flos-aquae. PMID- 16356523 TI - Complete nucleotide sequence and genomic organization of hepatopancreatic parvovirus (HPV) of Penaeus monodon. AB - We have determined the genome of hepatopancreatic parvovirus (HPV), a minus, single-stranded DNA virus isolated from infected Penaeus monodon in Thailand. Its genome consisted of 6321 nucleotides, representing three large open reading frames (ORFs) and two non-coding termini. The left (ORF1), mid (ORF2), and right (ORF3) ORFs on the complementary (plus) strand may code for 428, 579, and 818 amino acids, equivalent to 50, 68, and 92 kDa, respectively. The 5' and 3' ends of viral genome contained hairpin-like structure length of approximately 222 and 215 bp, respectively. No inverted terminal repeat (ITR) was detected. The ORF2 contained conserved replication initiator motif, NTP-binding and helicase domain similar to NS-1 of other parvoviruses. Therefore, it most likely encoded the major nonstructural protein (NS-1). The ORF1 encoded putative nonstructural protein-2 (NS-2) with unknown function. The ORF3 of the HPV genome encoded a capsid protein (VP) of approximately 92 kDa. This may be later cleaved after arginine residue to produce a 57-kDa structural protein. A phylogenetic tree based on conserved amino acid sequences (119 aa) revealed that it is closely related to Brevidensoviruses, which are shrimp parvovirus (IHHNV) and mosquito densoviruses (AaeDNV and AalDNV). However, the overall genomic organization and genome size of HPV were different from these parvoviruses, for instance, the non overlapping of NS1 and NS2, the larger VP gene, and the bigger genome size. This suggested that this HPV virus is a new type in Parvoviridae family. We therefore propose to rename this virus P. monodon densovirus (PmDNV). PMID- 16356524 TI - Crystal structure of the serine protease domain of Sesbania mosaic virus polyprotein and mutational analysis of residues forming the S1-binding pocket. AB - Sesbania mosaic virus (SeMV) polyprotein is processed by its N-terminal serine protease domain. The crystal structure of the protease domain was determined to a resolution of 2.4 A using multiple isomorphous replacement and anomalous scattering. The SeMV protease domain exhibited the characteristic trypsin fold and was found to be closer to cellular serine proteases than to other viral proteases. The residues of the S1-binding pocket, H298, T279 and N308 were mutated to alanine in the DeltaN70-Protease-VPg polyprotein, and the cis-cleavage activity was examined. The H298A and T279A mutants were inactive, while the N308A mutant was partially active, suggesting that the interactions of H298 and T279 with P1-glutamate are crucial for the E-T/S cleavage. A region of exposed aromatic amino acids, probably essential for interaction with VPg, was identified on the protease domain, and this interaction could play a major role in modulating the function of the protease. PMID- 16356525 TI - Chimpanzee adenovirus vaccine protects against Zaire Ebola virus. AB - This study evaluated the use of a chimpanzee-based adenovirus vaccine in mouse and Guinea pigs models of Zaire Ebola virus (ZEBOV) infection. Vaccine vector expressing the envelope glycoprotein of ZEBOV was created from the molecular clone of chimpanzee adenovirus pan7 (AdC7). AdC7 vaccine stimulated robust T and B cell responses to ZEBOV in naive mice inducing complete protection to an otherwise lethal challenge of ZEBOV. Complete protection to Zaire Ebola virus was also observed in Guinea pigs vaccinated with a relatively low dose of AdC7 (5 x 10(9)/kg). Pre-existing immunity to AdHu5 was generated in mice following pre exposure to AdHu5 or administration of pooled human immune globulin. Pre-existing immunity to human adenoviruses severely compromised the efficacy of the human AdHu5 vaccine but not the chimpanzee AdC7 vaccine. These results validate further development of Chimpanzee-based vaccine and highlight the impact of pre-existing immunity to the vaccine carrier. PMID- 16356526 TI - A reversed structure-from-motion effect for simultaneously viewed stereo surfaces. AB - A spatially flat stimulus is perceived as varying in depth if its velocity structure is consistent with that of a three-dimensional (3D) object. This is structure from motion (SFM). We asked if the converse effect also exists. A motion-from-structure effect would skew an object's perceived velocity structure to make it more consistent with the 3D structure provided by its depth cues. This proposed phenomenon should be opposite in sign from velocity constancy and could potentially interfere with it. Previous tests of velocity constancy compared stimuli presented at different times, not simultaneously. This explains why a reversal of SFM has not been previously reported, as it is expected to appear only for simultaneous presentations. We tested this prediction using random-dot stereograms to define two adjacent moving surfaces separated in stereoscopic depth. We found that subjects did not perceive velocity constancy with either simultaneous or sequential stimulus presentations. For sequential presentations, subjects matched retinal speeds, in agreement with previous work. However, for simultaneous presentations, the nearer surface was seen as moving faster when both surfaces were moving with the same retinal speed, an effect opposite in polarity from velocity constancy and a signature of the motion-from-structure phenomenon. PMID- 16356527 TI - Shape analysis of female facial attractiveness. AB - Previous studies have suggested that female facial attractiveness is associated with exaggerated sex-specific facial traits and averageness. Here we applied geometric morphometrics, a method for multivariate statistical analysis of shape, to measure geometric averageness and geometric sexual dimorphism of natural female face profiles. Geometric averageness and geometric sexual dimorphism correlate with attractiveness ratings. However, principal component analysis extracted a shape component robustly correlated with attractiveness but independent of sexual dimorphism. The shape differences between attractive- and hyperfeminine traits are localised: attractive facial shape and sexual dimorphism are similar in the upper face, but are markedly distinct in the jaw and chin. PMID- 16356528 TI - Peripheral refraction along the horizontal and vertical visual fields in myopia. AB - Peripheral refractions were measured to 35 degrees eccentricity using a free space autorefractor in young adult emmetropic and myopic subjects. Refractions were measured along horizontal and vertical visual fields for 116 subjects and a 43 subject subset, respectively. Along the horizontal visual field, peripheral myopic shifts in spherical equivalent M of emmetropes changed to relative hypermetropic shifts in the myopes, there were temporal-nasal asymmetries of 90 degrees to 180 degrees astigmatism J(180) which decreased as myopia increased, and 45 degrees to 135 degrees astigmatism J(45) was linearly related to field angle. Along the vertical visual field, both peripheral myopic shifts in peripheral M and J(180) asymmetry were unaffected by magnitude of myopia, and J(45) changed at three times the rate as for the horizontal visual field. Myopia has more effect on peripheral refraction of adult eyes along the horizontal than along the vertical visual field. The peripheral variations in refraction match well what is known about the shapes of emmetropic and myopic eyes. PMID- 16356529 TI - The visual motion detectors underlying ocular following responses in monkeys. AB - Psychophysical evidence indicates that visual motion can be sensed by low-level (energy-based) and high-level (feature-based) mechanisms. The present experiments were undertaken to determine which of these mechanisms mediates the initial ocular following response (OFR) that can be elicited at ultra-short latencies by sudden motion of large-field images. We used the methodology of Sheliga, Chen, Fitzgibbon, and Miles (Initial ocular following in humans: A response to first order motion energy. Vision Research, 2005a), who studied the initial OFRs of humans, to study the initial OFRs of monkeys. Accordingly, we applied horizontal motion to: (1) vertical square-wave gratings lacking the fundamental ("missing fundamental stimulus") and (2) vertical grating patterns consisting of the sum of two sinusoids of frequency 3f and 4f, which created a repeating pattern with beat frequency, f. Both visual stimuli share a critical property: when subject to 1/4 wavelength steps, their overall pattern (feature) shifts in the direction of the steps, whereas their major Fourier component shifts in the reverse direction (because of spatial aliasing). We found that the initial OFRs of monkeys to these stimuli, like those of humans, were always in the opposite direction to the 1/4 wavelength shifts, i.e., in the direction of the major Fourier component, consistent with detection by (low-level) oriented spatio-temporal filters as in the well-known energy model of motion analysis. Our data indicate that the motion detectors mediating the initial OFR have quantitatively similar properties in monkeys and humans, suggesting that monkeys provide a good animal model for the human OFR. PMID- 16356530 TI - Motion in depth from interocular velocity differences revealed by differential motion aftereffect. AB - There are two possible binocular mechanisms for the detection of motion in depth. One is based on disparity changes over time and the other is based on interocular velocity differences. It has previously been shown that disparity changes over time can produce the perception of motion in depth. However, existing psychophysical and physiological data are inconclusive as to whether interocular velocity differences play a role in motion in depth perception. We studied this issue using the motion aftereffect, the illusory motion of static patterns that follows adaptation to real motion. We induced a differential motion aftereffect to the two eyes and then tested for motion in depth in a stationary random-dot pattern seen with both eyes. It has been shown previously that a differential translational motion aftereffect produces a strong perception of motion in depth. We show here that a rotational motion aftereffect inhibits this perception of motion in depth, even though a real rotation induces motion in depth. A non horizontal translational motion aftereffect did not inhibit motion in depth. Together, our results strongly suggest that (1) pure interocular velocity differences can produce motion in depth, and (2) the illusory changes in position from the motion aftereffect are generated relatively late in the visual hierarchy, after binocular combination. PMID- 16356531 TI - Thermodynamics of metal cation binding by a solid soil derived humic acid. 2. Binding of Mn(II), Co(NH3)6aq3+ and Hg(II). AB - Metal binding is an important function of humic acids (HAs) in soils, sediments and waters. At pH 2.0, Mn(II) and Co(NH3)6aq3+ bind tightly in one step labeled A to a solid humic acid NHA isolated from a New Hampshire soil. Two consecutive steps are observed for Hg(II) binding. All the binding isotherms fit the Langmuir model in the temperature range 10.0-50.0 degrees C. Stoichiometric site capacities indicate predominant binding by charge-neutralizing HA carboxylate groups for Mn(II) and the second step A of Hg(II) binding. The binding affinity order in step A is Co(NH3)(6)3+>Hg(II)>Mn(II). Metal binding enthalpy and entropy changes fit the linear correlation found previously for binding of other metal cations by solid HAs. Free energy buffering from cooperative enthalpy and entropy changes and lower enthalpies for metal-HA interactions in solution suggest that desolvation of the cations and HA binding sites as well as HA conformational changes to allow for inner-sphere complexation predominate metal binding by hydrated solid HAs. PMID- 16356532 TI - Developmental effects in Japanese medaka (Oryzias latipes) exposed to nonylphenol ethoxylates and their degradation products. AB - The endocrine modulating potency of five alkylphenol compounds to fish, including nonylphenol (NP), three nonylphenol ethoxylate mixtures (NP1EO, NP4EO, NP9EO) and one nonylphenol ethoxycarboxylate (NP1EC) was assessed using in vivo tests conducted with Japanese medaka (Oryzias latipes). Medaka exposed to test materials from 1 day to 100 days post-hatch were monitored for alterations to sex ratios and secondary sex characteristics and development of gonadal intersex (i.e., testis-ova). The treatment with 100 microg l-1 NP (measured 29 microg l-1) induced gonadal intersex in over 80% of exposed males, mixed secondary sex characteristics in over 40% of exposed fish and suppression of the development of papillae on the anal fin of 100% of males. The 30 microg l-1 NP (measured 8.7 microg l-1) treatment induced gonadal intersex in only one of the 22 exposed males and mixed secondary sex characteristics in approximately 20% of the exposed fish. An elevated incidence of fish with mixed secondary sex characteristics and suppression of papillae development was also observed in the treatment with NP1EO at the highest test concentration of 300 microg l-1 (measured 105 microg l-1). There was no evidence of mixed secondary sex characteristics or gonadal intersex in treatments with the remaining test mixtures. This study confirms that NP is an estrogenic compound that could affect gonadal development in fish chronically exposed to concentrations in the range of 10 microg l-1. NP1EO is very weakly estrogenic at concentrations that are an order of magnitude higher than the lowest observed effect concentration for nonylphenol. PMID- 16356533 TI - Exposure of arc-furnace-plant workers to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). AB - This study aimed to evaluate serum polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) levels in electric-arc-furnace workers according to their corresponding occupational exposure. In addition, the ambient PCDD/Fs of the work environment were measured to provide additional support for the exposure and accumulation inside the electric arc furnace. The ambient PCDD/F concentrations inside the electric arc furnace were 1.557-1.917 pg-TEQ/Nm(3), 5-24 folds higher than those outside (0.080-0.385 pg-TEQ/N m(3)). In addition, higher average serum levels were measured in the workers with high occupational dioxin exposure (24.0 pg WHO-TEQ/g lipid) than in those with lower occupational dioxin exposure (13.8 pg WHO-TEQ/g lipid). Higher PCDFs/PCDDs ratios were found in serum samples from high-exposure groups than in low-exposure groups. The higher ratio of PCDFs/PCDDs was also found in ambient samples consistent with other metallurgical processes reported previously. Our results suggest that PCDD/Fs exposure exits in the smelting process of electric arc furnace, and the occupational hygiene should be taken more seriously concern in that workplace. PMID- 16356534 TI - Tealeaves may release or absorb fluoride, depending on the fluoride content of water. AB - As the tea plant (Camellia sinensis) is known to accumulate fluoride from the soil, the tealeaves may contain high concentrations of fluoride, which is easily released during infusion. In this study, we have tested the possible effect of original fluoride concentration in the water on the fluoride release from tea. Moreover, we wanted to test the possible capacity of tealeaves (commercially available tea) to absorb fluoride from high-fluoride water. In low-fluoride water, fluoride is easily released from tealeaves. Depending upon the fluoride content of the water, dried tealeaves are able also to absorb fluoride. Thus, if a cup of tea is made from high-fluoride water, the fluoride concentration of the infusion may actually be lower than the original fluoride concentration of the water. PMID- 16356535 TI - Short-term variability of dissolved trace element concentrations in the Marne and Seine rivers near Paris. AB - The concentrations of dissolved trace elements (Li, B, Mn, Cu, As, Rb, Sr, Mo, Cd, Ba, Pb) in the Marne and Seine rivers in the Paris urban area were monitored over a 2-year period. The resulting data indicated moderate contamination of waters by the most toxic elements (Cu, As, Cd and Pb). The River Marne upstream and the River Seine downstream of the city of Paris displayed similar concentrations. However higher fluxes of trace elements were observed in the Seine than in the Marne due to their different discharges. Li, B, Rb, Sr and Ba concentrations were correlated with river discharge and concentrations were higher during high river flow. This was interpreted as a dilution by discharge from a major natural or anthropogenic source. Mn, Cu, Mo, Cd and Pb concentrations were not correlated with discharge. Dissolved Mn, Cu and Cd increased rapidly in summer, whereas the concentration of Mo decreased. These variations were attributed to redox processes. During summer when the dissolved oxygen concentrations decrease, Mn, Cu, Cd and Pb are released into solution whereas Mo is immobilised. Like metals, variations in arsenic contents were not linked with discharge. Its similarity with phosphate distribution suggests similar controls involving phytoplankton uptake and release from sediments through organic matter mineralization. PMID- 16356536 TI - Comment on Martinez-Garcia et al. "Heavy metals in human bones in different historical epochs". AB - Martinez-Garcia et al. (Sci. Tot Env. 348:51-72) have examined heavy metal exposure of humans in the Cartagena region using analysis of archaeological bones. An analysis of the lead and iron levels they report shows that they are physiologically implausible and must therefore result from diagenesis. This, and analogy with the known diagenetic origin of certain other elements, suggests that the other metal analyses they report are also unlikely to be in vivo concentrations. Lifetime heavy metal exposure cannot be deduced from diagenetically altered concentrations. PMID- 16356537 TI - Atmospheric gaseous mercury in Northern Taiwan. AB - The first attempt to systematically investigate the atmospheric mercury over the territory of Taiwan has been undertaken. Four sampling sites were selected in localities qualified as urban, suburban, industrial-rural and rural ones, within the northern part of Taiwan. The sites were monitored for atmospheric gaseous mercury alternately from September to December 2003 and from April to June 2004, by using a portable automated mercury analyzer Model Gardis-3. The results have shown rather high atmospheric gaseous mercury levels (up to 30 ng/m3 at the urban site, up to 22 ng/m3 at the rural site, up to 24 ng/m3 at the industrial-rural site, and up to 90 ng/m3 at the suburban site) which may indicate the presence of considerably strong point and area mercury sources whose origin still has to be detected more precisely. PMID- 16356538 TI - Estimating the pretest probability threshold to justify empiric administration of heparin prior to pulmonary vascular imaging for pulmonary embolism. AB - Expertly interpreted pulmonary vascular imaging (either ventilation-perfusion scan or computed tomography chest angiography) is not uniformly available at most hospitals, including those in the US. When evaluating a patient with suspected pulmonary embolism during times when pulmonary vascular imaging is not available, clinicians frequently face the decision of whether to administer heparin while awaiting availability of imaging. In this report, we analyze published data to quantitate the probability of death or disability from untreated pulmonary embolism versus the probability of serious bleeding for one, two and seven days of heparin therapy. For these three time points, we estimate the pretest probability of pulmonary embolism to justify the empiric administration of heparin. PMID- 16356539 TI - Circulating CD40 ligand is elevated only in patients with more advanced symptomatic peripheral arterial diseases. AB - INTRODUCTION: CD40 and its ligand participate in atherosclerosis formation, progression and destabilization. Increased soluble CD40 ligand (sCD40L) was observed in hypercholesterolemia, unstable angina and conditions with platelet activation. To date, there is no report on the association of sCD40L with angiographic peripheral artery disease (PAD) disease severity. MATERIALS AND METHODS: From March 1999 to April 2004, consecutive patients having angiographically documented PAD and given consents for pre-procedural serum sample use were recruited into this study. The key PAD lesions should be > or = 70% diameter stenotic at the lower limbs and patients were dichotomized into two groups depending on total PAD lengths. Peer angiographic control subjects were those free of coronary disease, PAD and major medical diseases. The serum samples were thawed and analyzed for sCD40L, monocyte chemotactic protein 1 (MCP-1) and hs-CRP in a single batch. RESULTS: A total of 63 well-defined lower-limb PAD patients and 30 control subjects were studied. Patients with PAD lengths >5 cm (N=38) presented higher sCD40L than those with lesions < or = 5 cm (N=25) (5430+/ 459 vs. 3889+/-507 pg/ml, p=0.037) and control subjects (5430+/-459 vs. 3973+/ 551 pg/ml, p=0.037). However, there was no significant difference in circulating MCP-1 (375+/-49 vs. 310+/-49 pg/ml and 297+/-24 pg/ml, respectively, p=0.371) or hs-CRP (0.64+/-0.16 vs. 0.51+/-0.15 mg/ml and 0.46+/-0.15 mg/dl, respectively, p=0.682) across three groups. PAD patients with associated coronary lesions did not differ in circulating CD40L, MCP-1 or hs-CRP from without and control subjects. CONCLUSIONS: Soluble CD40L was significantly elevated in patients with more advanced symptomatic PAD and might be an indicator for disease extent stratification. The distribution of sCD40L in PAD was not affected by coronary involvement or not. PMID- 16356540 TI - Detection of up-regulated genes in thrombin-stimulated human umbilical vein endothelial cells. AB - INTRODUCTION: Thrombin, a serine protease, plays an important role in such actions as coagulation, cell proliferation and inflammation. It has been sporadically reported that endothelial cells, when stimulated by thrombin via protease-activated receptors (PAR), express various mediators and proteins including cytokines, chemokines, growth factors, and adhesion molecules. However, the pleiotropic effect of thrombin on endothelial cells has not yet been fully elucidated. MATERIALS AND METHODS: We newly searched for the up-regulated genes in the thrombin-stimulated endothelial cells by thorough screening using a microarray chip, printed with 22,575 human genes, followed by verification using real-time PCR (n=3). RESULTS AND CONCLUSIONS: Twelve genes, which were 4.8 times or more up-regulated in a microarray analysis, were selected and further analyzed. In real-time PCR, ICAM-1, IL-8, BIRC3, COL3A1, CXCL3, and CXCL1 were significantly up-regulated in the thrombin-stimulated cells: 16.0-, 8.81-, 5.92-, 3.74-, 1.74-, and 1.66-fold, respectively. VCAM-1, CXCL2, CCL20, CSF2, CD69, and CCL2 were up-regulated in the thrombin-stimulated cells: 12.2-, 2.44-, 1.90-, 1.82-, 1.62-, and 1.06-fold, respectively, without attaining statistical significance. We demonstrated, for the first time, that BIRC3 (anti-apoptotic protein), COL3A1 (matrix protein synthesis), and CXCL3 (chemokine) were up regulated in the thrombin-stimulated HUVECs. PMID- 16356541 TI - Meal patterns and body weight after nicotine in male rats as a function of chow or high-fat diet. AB - Studies of the effects of nicotine (NIC) on meal patterns in rats often employ chow pellet diets that contain little fat, whereas humans using NIC commonly consume diets relatively rich in fat. The aim of the present study was therefore to compare the impact of NIC administration and NIC cessation on meal pattern in adult male rats offered a standard powdered chow (CHOW: 10.9% fat by calories) diet or a palatable high-fat (HIFAT: 58.3% fat by calories) diet. Computerized meal pattern analyses were conducted for male rats treated for 14 days with injections of either saline or 1.4 mg/kg/day of NIC (as the free base given in 5 equal amounts) during the dark phase and continued for 10 days after NIC cessation. The suppression of daily caloric intake by NIC was larger in HIFAT-NIC rats than in CHOW-NIC rats (p < .01), such that NIC induced a greater suppression of body weight in HIFAT-NIC rats, relative to CHOW-NIC rats (p < 0.02). NIC administration reduced MS in both CHOW and HIFAT rats. CHOW fed rats showed a gradual increase in meal number in response to NIC, whereas HIFAT fed rats showed a significant initial suppression of meal number, which returned to control levels by day 4 of the 14 day NIC treatment period. In addition, NIC increased water intake more in HIFAT fed rats than in CHOW rats. Cessation of NIC resulted in transient increases in daily caloric intake in CHOW and in HIFAT rats. The present study demonstrates that NIC actions on food intake suppression, meal patterns, and weight reduction differ depending on whether the rats are fed low- or high-fat diets. PMID- 16356542 TI - Comparison of pregnancy rates in dromedary camels (Camelus dromedarius) after deep intra-uterine versus cervical insemination. AB - The ovarian follicular wave patterns of sixty adult female camels were monitored by serial trans-rectal ultrasound examinations and when the dominant follicle reached 1.3-1.8 cm in diameter they received a single intravenous injection of 20 microg buserelin, to induce ovulation, and were inseminated with a known number of spermatozoa 24 h later. Ejaculates were collected from the male camels and diluted 1:1 in Green Buffer with 20% egg yolk (v:v) added. Sperm concentration and motility were assessed and a dose of 40, 80 or 150 x 10(6) motile spermatozoa were deposited either just through the cervix into the uterine body or at the tip of the horn ipsilateral with the ovary containing the dominant follicle. Insemination of 150, 80 and 40 x 10(6) spermatozoa into the uterine body resulted in conception rates of 53, 7 and 0%, respectively, whereas insemination at the tip of the uterine horn resulted in conception rates of 43, 40 and 7%, respectively. PMID- 16356543 TI - Expression of leptin ligand and receptor and effect of exogenous leptin supplement on in vitro development of porcine embryos. AB - The present study investigated the expression of ligand and receptor for leptin, and the effect of leptin supplementation on preimplantation development of porcine in vitro fertilized (IVF) and somatic cell nuclear transfer (SCNT) embryos. The IVF embryos were produced using frozen boar semen and SCNT embryos were obtained by nuclear transfer of fetal fibroblasts into enucleated oocytes. The protein expression of leptin ligand and receptor was investigated in in vitro matured oocytes, 2-, 4- and 8-cell embryos, morulae and blastocysts derived from IVF and SCNT using immunofluorescence. Both the ligand and receptor were detected in in vitro matured oocytes and all stage of IVF and SCNT embryos. The IVF and SCNT embryos were cultured in modified North Carolina State University (mNCSU)-23 medium supplemented with various concentrations (0, 1, 10, 100 or 1000 ng/mL) of leptin. The rates of cleavage at day 2 and blastocyst formation at day 7, and cell number of blastocysts were monitored as experimental parameters. In SCNT embryos, supplementing with 1000 ng/mL leptin significantly (P<0.05) increased the rate of blastocysts formation (20.2% versus 12.9%) and total cell number (54.6+/-17.4 versus 45.1+/-15.2) compared to the control group. In IVF embryos, leptin supplementation did not affect preimplantation embryo development and cell number in blastocysts. In conclusion, the present study demonstrated the expression of leptin ligand and receptor and the embryotropic effect of leptin in SCNT embryos. PMID- 16356544 TI - Expression, localisation and hormone regulation of the human copper transporter hCTR1 in placenta and choriocarcinoma Jeg-3 cells. AB - Copper is an essential trace element necessary for normal growth and development. During pregnancy, copper is transported from the maternal circulation to the fetus by mechanisms which have not been clearly elucidated. The copper uptake protein, hCTR1 is predicted to play a role in copper transport in human placental cells. This study has examined the expression and localisation of hCTR1 in human placental tissue and Jeg-3 cells. In term placental tissue the hCTR1 protein was detected as a 105 kDa protein, consistent with the size of a trimer which may represent the functional protein. A 95 kDa band, possibly representing the glycosylated protein, was also detected. hCTR1 was localised within the syncytiotrophoblast layer and the fetal vascular endothelial cells in the placental villi and interestingly was found to be localised toward the basal plasma membrane. It did not co-localise with either the Menkes or the Wilson copper transporting ATPases. Using the placental cell line Jeg-3, it was shown that the 35 kDa monomer was absent in the extracts of cells exposed to insulin, estrogen or progesterone and in cells treated with estrogen an additional 65 kDa band was detected which may correspond to a dimeric form of the protein. The 95 kDa band was not detected in the cultured cells. These results provide novel insights indicating that hormones have a role in the formation of the active hCTR1 protein. Furthermore, insulin altered the intracellular localisation of hCTR1, suggesting a previously undescribed role of this hormone in regulating copper uptake through the endocytic pathway. PMID- 16356545 TI - TRPV6 potentiates calcium-dependent cell proliferation. AB - The Ca(2+) homeostasis within cells controls a diversity of cellular processes including gene transcription, proliferation and apoptosis. Perturbance of Ca(2+) signaling may induce deregulation of cell proliferation and suppression of cell death providing the basis for cancer development. In human prostate cancer, a correlation between the mRNA expression of the Ca(2+) channel TRPV6 and the staging of the cancer has been described. We have analyzed the influence of TRPV6 on cell proliferation within HEK-293 cells. We show that TRPV6 increases cell proliferation of HEK-293 cells in a Ca(2+) dependent manner. The increased proliferation correlates with slightly increased intracellular Ca(2+) levels without interfering with the intrinsic Ca(2+) dependence of HEK-293 cell proliferation. Low doses of econazole inhibit both, TRPV6 dependent Ca(2+) signals and cell proliferation while BTP2, a potent inhibitor of Ca(2+) signals and cell proliferation in T-cells, neither influences TRPV6 dependent Ca(2+) signals nor cell proliferation of HEK-293 cells. Our data demonstrate that TRPV6 increases the rate of Ca(2+) dependent cell proliferation which is a prerequisite for its potential role in tumor progression. PMID- 16356546 TI - A multiparametric approach for monitoring immunotoxic responses in mussels from contaminated sites in Western Mediterranea. AB - As a part of the multidisciplinary program Biological Effects of Environmental Pollution in Marine Coastal Ecosystems of the European Commission, this study aimed to validate immunological alterations as biomarkers of exposure to chemical contamination in polluted areas of Western Mediterranea. The status of the immune system has been assessed in mussels (Mytilus galloprovincialis) by measuring several immunopathological and immunocompetence parameters. Alterations of total hemocyte counts, lysosomal stability, and phagocytosis were among the most reliable effects observed in polluted sites and suggested immunosuppressive conditions in contaminated mussels. An immunotoxicological index was calculated from the set of individual data. By providing a single value per sampling station to score immunological alterations in mussels, this novel approach allowed recognition of a gradient of perturbation correlated to pollution intensity in two of the three sites monitored. Processing a set of biological parameters by this method was found to increase the ecotoxicological relevance of such multiparametric studies for the assessment of chemical contamination in coastal waters. PMID- 16356547 TI - The dual origin hypothesis: an evolutionary brain-behavior framework for analyzing psychiatric disorders. AB - According to the dual origin hypothesis, the cerebral cortex of higher mammals evolved from two primordial brain structures, the amygdala and hippocampal formation. This developmental process defines the orderly principles of cortical connectivity and gives rise to functionally distinct ventral and dorsal systems within the cerebrum. This paper reviews the basic features of the dual origin theory. This model is then applied to understanding symptom production in a number of psychiatric illnesses, with particular reference to recent structural and functional imaging studies. In this paper I propose that psychiatric symptoms can be conceptualized as arising from abnormal processing within dorsal (time space-motility) or ventral (meaning-motivation) systems, or from a disturbance in the functional interaction/balance between them. Within this framework, one can identify symptom-specific correlations that cross-traditional diagnostic boundaries, as well as potential mechanisms that may explain biologically valid diagnostic entities. Integrating evolutionary, connectional and functional bases across multiple species, the dual origin hypothesis offers a powerful neural systems model to help organize our understanding of psychiatric illness, therein suggesting novel approaches to diagnosis, prevention and treatment. PMID- 16356548 TI - Relationship between the brain and aggression. PMID- 16356549 TI - The analysis of human health risk with a detailed procedure operating in a GIS environment. AB - An approach for quantifying the human health risk caused by industrial sources, which, daily or accidentally, emit dangerous pollutants able to impact on different environmental media, is introduced. The approach is performed by the HHRA-GIS tool which employs an integrated, multimedia, multi-exposure pathways and multi-receptors risk assessment model able to manage all the steps of the analysis in a georeferenced structure. Upper-bound excess lifetime cancer risk and noncarcinogenic hazards are the risk measures, the spatial distribution of which is calculated and mapped on the involved territory, once all the pathways and receptors of the study area are identified. A sensitivity analysis completes the calculations allowing to understand how risk estimates are dependent on variability in the factors contributing to risk. The last part of the paper makes use of a case study concerning a working industrial site to put in evidence in which way the designed tool can help local authorities and policy makers in managing risks and planning remedial and reduction actions. The considered geographical area is a hypothetical territory characterized by residential, agricultural and industrial zones. The presence of two sources of contamination, a municipal waste incinerator (MWI) and a contaminated site, are evaluated by the tool application. Various typologies of receptors have been taken into account, each of them characterized by different anatomical and dietary properties. The achieved results are analyzed, compared with acceptable and background values and alternatives of minor environmental impact calculated. PMID- 16356550 TI - Orientations of axially coordinated imidazoles and pyridines in crystal structures of model systems of cytochromes. AB - Many properties of cytochromes and model systems depend on orientations of axial ligands. In this work, we elucidated the role of porphyrin substituents on orientation of axial ligands in model systems of cytochromes. The orientations of axially coordinated imidazoles and pyridines in crystal structures of model systems of cytochromes were analyzed and data were compared with previous quantum chemical calculations. The results show that eight ethyl groups on porphyrin ring strongly favor parallel orientation, hence, in all these complexes axial ligands, pyridines or imidazoles, are mutually parallel. Four phenyl or mesityl groups at meso-carbons also favor parallel orientation but less strongly. Hence, in most of the bis-imidazole complexes the orientation is parallel, while in bis-pyridine complexes the orientation of pyridines depends on oxidation state of Fe. In bis pyridine Fe(II) complexes orientation is parallel, in Fe(III) it is orthogonal. This analysis is in agreement with previous quantum-chemical calculations. PMID- 16356551 TI - Large-conductance, calcium-activated potassium channels: structural and functional implications. AB - The large-conductance, calcium-activated potassium channels (BK, also termed BK(Ca), Slo, or MaxiK) distributed in both excitable and non-excitable cells are involved in many cellular functions such as action potential repolarization; neuronal excitability; neurotransmitter release; hormone secretion; tuning of cochlear hair cells; innate immunity; and modulation of the tone of vascular, airway, uterine, gastrointestinal, and urinary bladder smooth muscle tissues. Because of their high conductance, activation of BK channels has a strong effect on membrane potential. BK channels differ from all other potassium (K(+)) channels due to their high sensitivity to both intracellular calcium (Ca(2+)) concentrations and voltage. These features make BK channels ideal negative feedback regulators in many cell types by decreasing voltage-dependent Ca(2+) entry through membrane potential hyperpolarization. The current review aims to give a comprehensive understanding of the structure and molecular biology of BK channels and their relevance to various pathophysiological conditions. The review will also focus on the therapeutic potential and pharmacology of the various BK channel activators and blockers. PMID- 16356552 TI - Objective evaluation of manual performance deficits in neurological movement disorders. AB - Impaired hand function is a frequent finding in movement disorders. The skilled control of prehensile finger forces is an essential feature of tool use in daily life. In healthy subjects, grip force is precisely adjusted to the mechanical object properties, such as weight and surface friction. Grip force is accurately scaled to be only a small amount higher than the minimum necessary to prevent a hand-held object from slipping. When an object is lifted and moved around in space, grip force is modulated in parallel with the movement-induced fluctuations in load. The absence of a temporal delay between grip and load force profiles implies that the central nervous system is able to predict the load variations before the intended manipulation. Sensory information is used to adjust the level of applied finger forces efficiently to the requirements of the mechanical object properties and the task at hand. The characteristics of impaired finger force control include inefficient grip force scaling and imprecision of the temporal coupling between grip and load force profiles. Here, we review the characteristics of deficient grip force behavior in movement disorders, e.g. Parkinson's disease, task-specific dystonia, Gille de la Tourette's syndrome and cerebellar disease. Grip force analysis is a highly sensitive method to document even subtle impairments of finger force control and may be used both as a diagnostic tool and for the objective evaluation of treatment in neurological movement disorders. PMID- 16356553 TI - Analysis of COMT gene (Val 158 Met polymorphism) in the clinical response to SSRIs in depressive patients of European origin. AB - INTRODUCTION AND OBJECTIVES: There is convincing evidence of interactions between serotonergic and dopaminergic systems and it seems that an increase of dopamine concentration in the whole brain could be a limiting factor for the antidepressant like effect of antidepressants. The COMT gene might be a good candidate for explaining some aspects of the pharmacological response to SSRIs. METHODS: The aim of our study was to analyse the Val 158 Met functional polymorphism on COMT gene and clinical response (4 weeks) and clinical remission (6/8 and 12 weeks) in two samples of depressive patients (DSM-IV) treated with SSRIs of Italian and Spanish origin. Clinical outcome was measured using 21 items Hamilton scale, weekly in the Italian sample (along 6 weeks) and monthly in the Spanish one (along 12 weeks). RESULTS: No overall effect of genotype or genotypextime interaction was detected. However, we observed a genotypextime interaction on HDRS decrease for citalopram treatment (F((4.6,317.5)) = 3.38, P = 0.007) in the Spanish sample. No clear effect was observed in the Italian sample. The three samples were pooled in order to test if carrying the Met/Met genotype confers an increased risk for non-remission at 6-8 weeks. The results showed that Met/Met carriers have an odds ratio of 2.21 (95% CI [1.20-4.12]) for non remission (chi(2) = 7.43, df = 2, P = 0.006). The Met/Met effect was not observed in response at 4th week (for all SSRI treatments) or in remission at 12th week (citalopram treatment). CONCLUSIONS: COMT gene could have a small and indirect effect of clinical response to SSRIs by slowing-down the antidepressant action along the follow-up, basically in citalopram treatment. PMID- 16356554 TI - Two dimensional Blue Native-/SDS-PAGE analysis of SLP family adaptor protein complexes. AB - SH2 domain containing leukocyte protein (SLP) adaptor proteins serve a central role in the antigen-mediated activation of lymphocytes by organizing multiprotein signaling complexes. Here, we use two dimensional native-/SDS-gel electrophoresis to study the number, size and relative abundance of protein complexes containing SLP family proteins. In non-stimulated T cells all SLP-76 proteins are in a approximately 400 kDa complex with the small adaptor protein Grb2-like adaptor protein downstream of Shc (Gads), whereas half of Gads is monomeric. This constitutive SLP-76/Gads complex could be reconstituted in Drosophila S2 cells expressing both components, suggesting that it might not contain additional subunits. In contrast, in B cells SLP-65 exists in a 180 kDa complex as well as in monomeric form. Since the complex was not found in S2 cells expressing only SLP-65, it was not di/trimeric SLP-65. Upon antigen-stimulation only the complexed SLP-65 was phosphorylated. Surprisingly, stimulation-induced alteration of SLP complexes could not be detected, suggesting that active signaling complexes form only transiently, and are of low abundance. PMID- 16356556 TI - Management of pediatric plunging ranula. AB - OBJECTIVE: There appears to be a high incidence of ranula in New Zealand. This study was designed to evaluate the diagnosis and surgical treatment of plunging ranula in children at Auckland Starship Children's Hospital, New Zealand. METHODS: A prospective study of 21 pediatric patients with a clinical diagnosis of plunging ranula was conducted. All children underwent clinical assessment and fine needle aspiration cytology. Cytological diagnosis was considered definitive and radiological investigations were not conducted. Surgical treatment comprising intraoral excision of the ipsilateral sublingual gland and pseudocyst was performed. RESULTS: The preoperative diagnosis was unchanged after surgery and histopathological analysis in all cases. There were no recurrences at 24-month follow up. There are no long-term complications to date. CONCLUSIONS: Intraoral removal of the sublingual glands and pseudocyst is an effective and safe method for the treatment of plunging ranula in the pediatric population. PMID- 16356555 TI - Neuronal surface glycolytic enzymes are autoantigen targets in post-streptococcal autoimmune CNS disease. AB - Infection with the Group A Streptococcus (GAS) can result in immune mediated brain disease characterised by a spectrum of movement and psychiatric disorders. We have previously described anti-neuronal antibodies in patients that bind to a restricted group of brain antigens with molecular weights 40 kDa, 45 kDa (doublet) and 60 kDa. The aim of this study was to define these antigens using 2 dimensional electrophoresis or ion exchange and hydrophobic interaction chromatography, followed by mass spectrometry. The findings were confirmed using commercial antibodies, commercial antigens and recombinant human antigens. The autoantigens were neuronal glycolytic enzymes--NGE (pyruvate kinase M1, aldolase C, neuronal-specific and non-neuronal enolase). These are multifunctional proteins that are all expressed intracellularly and on the neuronal cell surface. On the neuronal plasma membrane, NGE are involved in energy metabolism, cell signalling and synaptic neurotransmission. Anti-NGE antibodies were more common in the 20 unselected post-streptococcal CNS patients compared to 20 controls. In vitro experiments using cultured neurons showed that commercial anti-NGE antibodies induced apoptosis compared to blank incubation and control anti-HuD antibody. GAS also expresses glycolytic enzymes on cell surfaces that have 0-49% identity with human NGE, suggesting molecular mimicry and autoimmune cross reactivity may be the pathogenic mechanism in post-streptococcal CNS disease. PMID- 16356557 TI - Amelioration of cognitive impairment and changes in microtubule-associated protein 2 after transient global cerebral ischemia are influenced by complex environment experience. AB - In this study we examined whether expression of microtubule-associated protein 2 (MAP2) after transient global cerebral ischemia can be influenced by behavioral experience and if the changes are associated with functional improvement. Rats received either ischemia or sham surgery then assigned to: complex environment housing (EC) or social housing (SC) as controls for 14 days followed by water maze testing. Upregulation of MAP2 was seen in all ischemic animals with a significant overall increase evident in the EC housed rats. Behaviorally, all animals learned to perform the water maze task over time but the ischemia SC rats had the worst performance overall while all the EC housed animals demonstrated the best performance in general. Regression analysis showed that increase MAP2 expression was able to explain some of the variance in the behavioral parameters in the water maze suggesting that this cytoskeletal protein probably played a role in mediating enhanced functional outcomes. PMID- 16356558 TI - The Concentric Square Field: a multivariate test arena for analysis of explorative strategies. AB - In this study, we describe the behavior of laboratory rats in a recently developed observation arena, the Concentric Square Field (CSF). The CSF contains a number of areas designed to provoke exploration and behaviors associated with risk assessment, risk taking and security seeking in an environment not previously experienced. The model includes sheltered, open and elevated areas, a hole board device, areas with different light conditions, and wall-enclosed corridors. The rationale behind the CSF is to meet the demand for multivariate test situations that are not predictive in the sense of previous definition of a specific purpose of measuring a certain mental state. We define multivariate as being a free choice of where to stay in areas of different qualities. In the present study, identification of risky as opposed to safe areas is based on the retrieval behavior in lactating females and hoarding of food pellets in food deprived males. Furthermore, we describe the effects of pre-trial food deprivation, immobilization, social stress, strain differences (Sprague-Dawley, Wistar and Lister Hooded males), sex differences (Sprague-Dawley) and repeated testing. Besides the conventional statistics, a principal component analysis (PCA) helped to discriminate between the various categories tested. Our conclusion is that the multivariate and non-predictive test situation (CSF) and the use of PCA provide a good tool for ethoexperimental analysis. PMID- 16356559 TI - Physiological changes and differential gene expression in mummichogs (Fundulus heteroclitus) exposed to arsenic. AB - Arsenic has been detected as a contaminant in water bodies around the world. Although a number of studies have shown toxicity to adult fish, little is known about its effects on the offspring. However, human epidemiological studies have shown that arsenic increases the number of stillbirths and prematurely born infants. We examined changes in the morphology and gene expression in juvenile mummichogs (Fundulus heteroclitus) whose parents were exposed to 230 ppb arsenic for 10 days immediately prior to spawning. The hatchlings of exposed fish had a 2.8-fold increased incidence of curved or stunted tails. Total RNA from 6-week old hatchlings, reared in clean water, was used to construct a cDNA subtractive hybridization library. Using this library, we found 13 genes whose expression was altered in the hatchlings as a result of arsenic exposure. We confirmed differential expression by real-time PCR and found significant up-regulation of myosin light chain 2 (4.2-fold), type II keratin (1.5-fold), tropomyosin (3.1 fold) and parvalbumin (3.5-fold) in the hatchlings whose parents were exposed to arsenic. These genes are important during embryogenesis and their differential expression may be linked to the morphological changes observed in the hatchlings. PMID- 16356560 TI - Dependency of copper toxicity to polychaete larvae on algal concentration. AB - Algae are often used as food in aquatic metal toxicity tests to maintain the well being of test animals. Such food addition may change metal bioavailability because of the reduction of aqueous metal concentration and the increase in particle-bound metal concentration. While the importance of aqueous exposure pathway is widely recognized, few studies have determined the contribution of the dietary pathway to the overall metal toxicity to aquatic invertebrates. In this study, we determined the toxicity of both algal-bound copper alone and copper solution containing algae to the larvae of marine polychaete Hydroides elegans. Algae that had been pre-exposed to copper at up to 1024 microg l(-1), when fed to the larvae, did not cause significant abnormal larval development. However, when larvae were exposed to algal-copper mixture, percentage of normal larvae could be modeled as a logistic function of aqueous copper concentration, with a 48-h EC(50) (mean +/- S.E.) of 64.9 +/- 4.8 microg Cul(-1). When the toxicity was expressed using total copper concentration, the EC(50) ranged from 58.4 +/- 4.5 microg l(-1) in the control to 121.9 +/- 9.9 microg l(-1) in the 10(6) cells ml( 1) algal treatment. This study highlights the dependency of copper toxicity on the aqueous exposure pathway in this polychaete and the importance of considering algal binding of the metal in larval toxicological tests. PMID- 16356561 TI - Functional outcomes of elderly patients after the first hospital admission for decompensated heart failure (HF). A prospective study. AB - Congestive HF is one of the most common discharge medical diagnoses in elderly hospitalized patients. We evaluate prospectively the usefulness of a global geriatric assessment to identify changes in the functional status of patients who experience their first hospitalization for a new diagnosis of HF. The Barthel Index (BI), the Older Americans Resource Scale (OARS), the Short Portable Mental Status Questionnaire (SPMSQ), and the short form of the Mini Nutritional Assessment (short-MNA) were used to estimate functional, cognitive, and nutritional status. The Charlson score (CS) was used to measure comorbidity. Eighty-eight patients (mean age 79 years; 57% women) were finally included; their median CS score was 2.1. Prior to the index admission, their mean BI score was 91, OARS 9.8, SPMSQ 1.8 errors, and short-MNA 10.7. Twenty-four patients (27%) died during the first year of follow-up. Low preadmission BI scores were predictive of mortality (p=0.02), but not of readmission (p=0.9). After one-year of follow-up BI scores remained lower than preadmission values in 64% of the surviving patients; for OARS scores the figure was 67%. In conclusion, a previous low functional capacity is associated with higher mortality but not with HF related hospital readmission. Admission because of a new onset HF is often followed by a sustained functional decline both for the performance of basic and instrumental activities of the daily living. PMID- 16356562 TI - Acute phase reactants predict mitral regurgitation following mitral valvuloplasty. AB - This report evaluated whether acute phase reactants can predict the development of mitral regurgitation following percutaneous mitral valvotomy. 58 patients who developed significant mitral regurgitation following valvotomy were retrospectively compared with 58 age, sex and procedure technique matched control patients, who had valvotomy without mitral regurgitation. ESR and total leucocyte count were significantly higher in the group who developed mitral regurgitation, than in the control group. Higher ESR and total leucocyte count may be indicative of ongoing low grade sub-clinical inflammatory process, which makes the valve tissue friable which can give way during balloon stretch and lead onto mitral regurgitation. PMID- 16356563 TI - Aneurysm of the membranous interventricular septum demonstrated by multislice computed tomography. PMID- 16356564 TI - The relationship between variables of 123-I-metaiodobenzylguanidine cardiac imaging and clinical status of the patients with diastolic heart failure. AB - BACKGROUND: The status of cardiac sympathetic nerve activity in patients with diastolic heart failure has not been fully understood. 123-I metaiodobenzylguanidine cardiac images are valuable for evaluating cardiac sympathetic nerve activity. METHODS: We obtained 123-I-metaiodobenzylguanidine cardiac images from 34 consecutive patients with moderate heart failure and an ejection fraction of > or = 45%. RESULTS: The decay-corrected washout rate of 123 I-metaiodobenzylguanidine correlated with each plasma concentration of brain natriuretic peptide (standardized correlation coefficient=0.305, p<0.05), New York Heart Association functional class (standardized correlation coefficient=0.364, p<0.02), and exercise capacity (standardized correlation coefficient=-0.388, p<0.04). A multiple regression analysis revealed that the washout rate independently predicted plasma concentration of brain natriuretic peptide (standardized regression coefficient=0.367, p<0.02). In a univariate regression, the washout rate did not significantly correlate with the presence of ischemic heart disease (p=0.254); in a multivariate regression, the presence of ischemic heart disease did not predict the washout rate. For the 14 patients with sinus rhythm, there was a marginal negative correlation between the E/A velocity ratio of the transmitral flow and washout rate (standardized correlation coefficient=-0.518, p<0.07). CONCLUSIONS: In diastolic heart failure, cardiac sympathetic nerve activity increases proportionally to severity of the disease. PMID- 16356565 TI - Interaction of low serum adiponectin levels and smoking on coronary stenosis in Japanese men. AB - BACKGROUND: The relationship between serum adiponectin levels and coronary stenosis and the interaction of low serum adiponectin levels and smoking on coronary stenosis have not been clarified. METHODS: We conducted a hospital-based case-control study to examine serum adiponectin levels and smoking in 86 male patients with coronary stenosis and 145 male controls, using multiple logistic regression analyses. Serum adiponectin levels were grouped into quartiles for the analysis of the relationship between serum adiponectin levels and coronary stenosis, and into dichotomy for the analysis of the interaction of low serum adiponectin levels and smoking status. RESULTS: Decreasing serum adiponectin levels were significantly associated with increased risk of coronary stenosis in a dose-dependent manner. As for the interaction of low serum adiponectin levels and smoking on coronary stenosis, adjusted odds ratios were 14.00 (95% Confidence Interval 5.45-37.48) among ever-smokers with low serum adiponectin levels, 3.84 (95% CI: 1.44-10.21) among ever-smokers with high serum adiponectin levels and 3.79 (95% CI: 1.38-10.45) among never-smokers with low serum adiponectin levels, compared with the reference group of never-smokers with high serum adiponectin levels. CONCLUSIONS: The interaction of low serum adiponectin levels and smoking on coronary stenosis was likely to be a synergic effect. This study suggests that the combination of low serum adiponectin levels and smoking is the most useful predictor for coronary stenosis among Japanese men. PMID- 16356566 TI - Estrogen decreases neointimal hyperplasia and improves re-endothelialization in pigs. AB - BACKGROUND: 17-Beta-estradiol inhibits smooth muscle cell proliferation and migration and accelerates endothelial cell repair in response to balloon injury. The aim of this study was to determine whether administration of estrogen is associated with decreased neointimal hyperplasia after stenting in the porcine model. METHODS AND RESULTS: Twenty-two farm female pigs 7 months old were oophorectomized and at the third day normal saline (11 pigs) or 20 mg of estradiol valerate (11 pigs), modified release, were administered intramuscularly. At the 10th day after oophorerectomy, 1 or 2 stainless steel stents were implanted in the right coronary artery in each pig, and at the 17th day, normal saline or 10 mg estradiol valerate, modified release, were administered intramuscularly in the two groups. Pigs were sacrificed 28 days after stent implantation and histomorphometric analysis of the coronary arteries was performed (20 stents from the estrogen and 18 stents from the control groups). In the estrogen group, neointimal proliferation area was 1.42+/-0.55 mm(2), whereas in the control group, 1.96+/-0.89 mm(2) (p=0.02). Area stenosis was 39+/-13% and 49+/-16% in the two groups, respectively (p=0.07). Re endothelialization was 2.67+/-0.34 and 2.22+/-0.46 in the two groups, respectively (p<0.010). The injury score was similar between the two groups. CONCLUSIONS: These data suggest that intramuscular administration of estrogen accelerates the endothelial cell repair in response to injury and reduces intimal hyperplasia in the porcine model. PMID- 16356567 TI - Short-term effects of low-dose atorvastatin on inflammatory status and lipid profiles in perimenopausal hypercholesterolemic, hypertriglyceridemic women. AB - The short-term and small-dose pleiotropic effects of atorvastatin and influence on sex steroid production were investigated in 35 premenopausal and 71 postmenopausal hypercholesterolemic, hypertriglyceridemic women, as well as the temporal differences in these pleiotropic effects. Atorvastatin (10 mg daily) was given for 6 months and fasting lipid concentrations, high sensitive CRP, and coagulo-fibrinolytic parameters were measured at baseline and after 3 and 6 months of therapy. Atorvastatin reduced the low-density lipoprotein cholesterol, remnant-like particle lipoprotein cholesterol, and malondialdehyde-modified low density lipoprotein cholesterol after 3 and 6 months in both pre- and postmenopausal women. Atorvastatin decreased significantly high-sensitivity C reactive protein concentration (-47.6% and -58.0%, P<0.01) and tissue plasminogen activator/plasminogen activator inhibitor-1 ratio (-31.8% and -40.0%, P<0.001) after 6 months in pre- and postmenopausal women. There was no correlation between the pleiotropic effects and the improvement in the lipid profile. Furthermore, atorvastatin has no influence on sex steroid production in both pre- and postmenopausal period. The results indicate some short-term pleiotropic effects of small-dose atorvastatin therapy without influence of endocrinological status, which may be important with respect to the early benefits of statin therapy in the perimenopausal hyperlipidemic women. PMID- 16356568 TI - The value of changes in QRS width and in ST-T segment during exercise test in hypertrophic cardiomyopathy for identification of associated coronary artery disease. AB - OBJECTIVES: Non-invasive methods cannot reliably predict the presence of coronary artery disease (CAD) in hypertrophic cardiomyopathy (HCM). This study aims to define the accuracy of QRS width changes versus standard ST-T criteria for recognition of associated ischemic CAD in patients with HCM undergoing exercise testing (ET). METHODS: A retrospective study including patients with HCM. HCM was defined by left ventricular hypertrophy (LVH) of unknown etiology of at least 15 mm. Coronary angiography was performed as a gold standard for definition of CAD (> or =70% obstruction in at least one major artery). QRS width duration was measured at peak ET by a computerized method employing an optical scanner. No changes in QRS width or shortening during ET were considered normal; QRS width prolongation of more than 3 ms was defined as abnormal. RESULTS: 68 patients (56/12 M/F) aged 60+/-12 y were studied. During ET, abnormal QRS response was found in 40 (58.8%) and Ischemic ST-T changes in 52 (76.5%) patients. CAD in at least one artery was diagnosed in 31 patients (45.5%). The sensitivity of QRS width versus ST-T changes during ET for associated CAD was 82% and 28%, respectively. Specificity was 75% and 48%, respectively. Positive and negative predictive values were 88%; 68% for QRS width and 67%; 59% for ST-T changes respectively. CONCLUSIONS: In patients with HCM undergoing ET, the association with CAD was more accurately predicted by an increase in QRS complex width than by standard criteria of ST-T segment changes. Thus, its use should be encouraged, especially in patients with HCM. PMID- 16356569 TI - Exercise training increases myocardial inotropic response in food restricted rats. AB - This study evaluated the effects of exercise training on myocardial function and ultrastructure of rats submitted to different levels of food restriction (FR). Male Wistar-Kyoto rats, 60 days old, were submitted to free access to food, light FR (20%), severe FR (50%) and/or to swimming training (one hour per day with 5% of load, five days per week for 90 days). Myocardial function was evaluated by left ventricular papillary muscle under basal condition (calcium 1.25 mM), and after extracellular calcium elevation to 5.2 mM and isoproterenol (1 microM) addition. The ultrastructure of the myocardium was examined in the papillary muscle. The training effectiveness was verified by improvement of myocardial metabolic enzyme activities. Both 20% and 50% food restriction protocols presented minor body and ventricular weights gain. The 20%-FR, in sedentary or trained rats, did not alter myocardial function or ultrastructure. The 50%-FR, in sedentary rats, caused myocardial dysfunction under basal condition, decreased response to inotropic stimulation, and promoted myocardial ultrastructural damage. The 50%-FR, in exercised rats, increased myocardial dysfunction under basal condition but increased response to inotropic stimulation although there was myocardial ultrastructural damage. In conclusion, the exercise training in severe restriction caused marked myocardial dysfunction at basal condition but increased myocardial response to inotropic stimulation. PMID- 16356570 TI - How far to the hospital? The effect of hospital closures on access to care. AB - Do urban hospital closures affect health care access or health outcomes? We study closures in Los Angeles County between 1997 and 2003, through their effect on distance to the nearest hospital. We find that increased distance to the closest hospital increases deaths from heart attacks and unintentional injuries. This finding is robust to several sensitivity checks. We also find that, for residents with health insurance, increased distance shifts regular care towards doctor's offices. While most residents are otherwise unaffected, we find some evidence that seniors perceive more difficulty accessing care. PMID- 16356571 TI - A versatile and sensitive tritium-based radioassay for measuring hydrogenase activity in aquatic sediments. AB - We present a method for the measurement of hydrogenase (H(2)ase) activity in aquatic sediments. The assay is based on the H(2)ase-mediated isotopic exchange between dissolved molecular hydrogen (H(2)) and water. A slurry of sediment material is incubated with a tritiated hydrogen (HT) headspace in a glass syringe on a rotary shaker. The method includes a procedure for preparing HT from radiolabeled sodium borohydride, which is a useful alternative to purchasing HT directly. A method for measuring HT specific activity based on liquid scintillation counting is also presented. Validation tests were run using live and frozen cultures of Clostridium pasteurianum and Desulfovibrio vulgaris, and freshly collected marine sediments. Adherence to Michaelis-Menten kinetics was demonstrated. An interassay coefficient of variation of 15% was determined using frozen C. pasteurianum cultures as reference material. Serial dilutions of cultures and sediments showed that measured H(2)ase activity scales with cell concentration, and indicate that the method can detect C. pasteurianum cell concentrations of between 300 and 3000 cells/ml. This technique allows measurement of H(2)ase activity in a variety of environmental samples, and will be particularly useful in the study of deep marine sediments with low microbial activity. PMID- 16356572 TI - ERP topography and human perceptual learning in the peripheral visual field. AB - We studied human perceptual learning in the peripheral visual field in 16 healthy adults. Horizontal or vertical vernier stimuli were presented simultaneously at 8 locations at an eccentricity of 4 degrees . One of the stimuli displayed an offset, and subjects were asked to detect the target offset. Training was performed with either vertical or horizontal stimuli by the repeated presentation of stimuli. Discrimination performance was also measured with the untrained stimuli. Before and after the psychophysical experiment, EEG was recorded from 30 electrodes over the occipital areas (between the inion and Cz) while targets were presented at all locations as vernier onset/offset stimuli. The EEG was averaged for each orientation separately. Improvement in discrimination performance was observed in about 70% of the subjects with the trained orientation only. The evoked potential maps displayed three components occurring between 80 and 160, 180 and 260, and 280 and 340 ms. The potential field topography of the first and third component showed significant differences before and after learning. In addition, field strength (global field power) of the second and third component increased with learning. No effects were seen with the untrained stimuli in the psychophysical and electrophysiological experiments. Our findings suggest that perceptual learning in the peripheral visual field is specifically related to neurophysiological changes induced by training, and it is not caused by unspecific changes of spatial attention. The changes of electrical brain activity reflect short-term plasticity related to human perceptual learning. PMID- 16356573 TI - Lipase-catalyzed synthesis of sorbitol-fatty acid esters at extremely high substrate concentrations. AB - Lipase-catalyzed synthesis of sorbitol-fatty acid esters was performed in eutectic media with extremely high substrate concentrations. Homogeneous eutectic melts of sorbitol and fatty acids of C6-C16 were prepared using an adjuvant mixture. Enhanced homogeneity of mixtures was confirmed by X-ray diffraction analysis. The substrate concentration was 3.63-6.67 M in the eutectic media, whereas in organic media the concentration was below 0.10 M. Esters were synthesized with an immobilized Candida antarctica lipase, and optimum conditions were analyzed. Compared to reactions in organic media, the initial reaction rate of ester synthesis and the overall productivity were significantly enhanced in eutectic media while the conversion yields were similar. Based on the kinetic analysis, highly viscous eutectic media were shown to influence the initial reaction rate and the apparent activation energy resulting in diffusion limitations. PMID- 16356574 TI - Design of high-throughput-compatible protocols for microencapsulation, cryopreservation and release of bovine spermatozoa. AB - With a rate exceeding 90% in cattle, artificial insemination (AI) is the prime reproduction technology in stock farming. AI success is expected to increase with extended persistence of sperms in utero. In order to enable controlled sperm release during artificial insemination we have designed two strategies for the automated microencapsulation of bovine spermatozoa in either alginate-Ca2+ or cellulose sulfate (CS)-poly-diallyldimethyl ammonium chloride (pDADMAC) capsules using standard encapsulation hardware. Animal protein- and citric acid-free sperm extenders and encapsulation protocols have been developed to ensure encapsulation compatible with sperm physiology. Bovine spermatozoa have showed high motility rates inside CS-pDADMAC-based capsules, were preserved by standard cryoconservation and rescued with high viability/motility following disintegration of the thawed capsules. CS-pDADMAC-based capsules break up within 72 h after addition of either purified cellulase or cellulase-filled alignate Ca2+ capsules. The controlled release, associated with the microencapsulation of bovine spermatozoa, may be a promising approach to increase the success rate of artificial insemination. PMID- 16356575 TI - Targeted drug delivery crossing cytoplasmic membranes of intended cells via ligand-grafted sterically stabilized liposomes. AB - In this study, we tested whether sterically stabilized liposomes (SSL) with surface ligands specific for the mu opioid receptor (MOR) can actively target MOR expressing cells. Dermorphin, a selective MOR agonist, was conjugated to DSPE PEG(3400) to obtain DSPE-PEG(3400)-dermorphin. Dermorphin-grafted SSL (dermorphin SSL) was prepared by thin-film rehydration-extrusion and post-insertion method. DSPE-PEG(3400)-dermorphin and dermorphin-SSL retained the affinity to MOR as determined by receptor binding assay using [(3)H]DAMGO, whereas plain SSL without surface ligands showed no binding to the receptor. Cellular uptake of cholesteryl BODIPY encapsulated dermorphin-SSL was studied by microplate spectrofluorometry as well as fluorescent and confocal microscopy. Significant fluorescence signal was observed inside CHO-hMOR cells after the treatment with dermorphin-SSL, indicative of MOR-mediated endocytosis. In contrast, no uptake of dermorphin-SSL was found in naive CHO cells or CHO-hDOR cells that lack MOR. Taken together, these results demonstrate that dermorphin-SSL delivery system is capable of targeting intracellular components of MOR-expressing cells. Such a system may be applied to carry pharmaceutical agents to achieve region-specific delivery of analgesics and/or to attenuate side effects associated with opioids. PMID- 16356576 TI - Painless electroporation with a new needle-free microelectrode array to enhance transdermal drug delivery. AB - A microelectrode array was designed to minimize the pain sensation of electroporation for enhancing transdermal drug delivery. The influence of the size of the electrode-skin contact area and of the distance between electrodes on the pain sensation was tested on human volunteers. The pain level decreased with the dimension of electrode-skin contact area and with inter-electrode distance. When both reached about 0.5 mm, the pain level was not perceptible even at the threshold of transdermal electroporation level of sixty electric pulses at 150 V, 1 ms at 1-10 Hz. An array of 11 x 11 alternately connected electrodes with 0.6 x 0.6 mm dimension was fabricated. The electric thresholds for effective drug delivery, using toluidine blue O as a marker on mouse skin, was found to be the same for microelectrode arrays as for larger electrodes and wider inter-electrode distances. In vivo transdermal electroporation using microelectrode array with 180 pulses of 150 V, 0.2 ms at 1 Hz, followed by 30 min methotrexate (MTX) occlusion increased more than 4 fold the systemic MTX level in mice. The results demonstrated the potential of painless delivery of significant amounts of chemotherapeutic agents through skin with the new electrode arrays in a clinical setting. PMID- 16356577 TI - Long term outcome and response to therapy of primary biliary cirrhosis-autoimmune hepatitis overlap syndrome. AB - BACKGROUND/AIMS: Whether primary biliary cirrhosis (PBC)-autoimmune hepatitis (AIH) overlap syndrome requires immunosuppressive therapy in addition to ursodeoxycholic acid (UDCA) is a controversial issue. METHODS: Seventeen patients with simultaneous form of strictly defined overlap were followed for 7.5 years. First-line treatment was UDCA alone (UDCA) in 11 and combination of immunosuppressors and UDCA (UDCA + IS) in 6. RESULTS: Characteristics at presentation were not significantly different between the 2 groups. In the UDCA + IS group (f-up 7.3 years), biochemical response in terms of AIH features (ALT<2ULN and IgG < 16 g/L) was achieved in 4/6 and fibrosis did not progress. In the UDCA group, biochemical response was observed in three patients together with stable or decreased fibrosis (f-up 4.5 years) whereas the eight others were non responders with increased fibrosis in four (f-up 1.6 years). Seven of these eight patients subsequently received combined therapy for 3 years. Biochemical response was obtained in 6/7 and no further increase of fibrosis was demonstrated. Overall, fibrosis progression in non-cirrhotic patients occurred more frequently under UDCA monotherapy (4/8) than under combined therapy (0/6) (P = 0.04). CONCLUSIONS: Combination of UDCA and immunosuppressors appears to be the best therapeutic option for strictly defined PBC-AIH overlap syndrome. PMID- 16356579 TI - Treatment of viral hepatitis in HIV-coinfected patients-adverse events and their management. AB - For the treatment of HBV/HIV-co-infection, study data on interferon-based therapy are very limited and insufficient to draw any specific conclusions. In contrast, data on HBV-polymerase inhibitors (lamivudine, adefovir, tenofovir) are available from controlled trials. Lamivudine is well tolerated and safe, however, development of HBV-resistance is frequent. Adefovir has a nephrotoxic potential and may at least theoretically induce antiretroviral resistance in HBV/HIV patients treated with adefovir. Tenofovir has gastrointestinal side effects, is associated with hypophospatemia, which has not induced serious osteopenia so far and may have a nephrotoxic potential. For HCV/HIV-co-infection pegylated interferon alpha plus ribavirin is standard of care. Flu-like symptoms, fatigue and depressive mood changes are frequent. In patients with a history of neurotic or minor depression initiation of treatment with antidepressants before the start of interferon-based therapy should be considered. Weight loss may be pronounced in individual cases. A marked decrease in absolute, but not relative CD4 +/- cells is the rule, but no relevant increase in opportunistic infection was observed, and anaemia (<10 g/dl) is reported in up to 30% of patients. Neutropenia (< 1,000 cells/microl) is observed in up to 50% of the patients. Adverse events specific to the HCV/HIV-patient population as compared to HCV-mono infected patients are the occurrence of hyperlactataemia/lactic acidosis and hepatic decompensation. PMID- 16356578 TI - Options for treatment of hepatitis C in HIV-infected persons. AB - Irrespective of whether a patient has HIV infection, the optimal treatment for hepatitis C virus (HCV) infection is peginterferon alpha and ribavirin. In both HIV-infected and uninfected persons, sustained virologic response (SVR) rates are higher for genotype 2 and genotype 3 HCV infection and for patients with lower pre-treatment HCV RNA levels. HIV-infection does not alter either the reality that persons who fail to achieve a 2log(10) reduction in HCV RNA level after 12 weeks of therapy rarely achieve a SVR, or the theoretical benefits of maintenance therapy in those without viral responses. The same adverse treatment effects can occur in HIV-infected and uninfected persons, but treatment of HIV-infected persons is complicated by interactions between ribavirin and antiretroviral medications and effects of HCV treatment on the course of HIV. The optimal treatment doses and durations are not known for HIV-infected persons, who are also less likely to achieve a SVR. A final difference is that the benefits of HCV treatment breakthroughs are usually realized in patients without HIV years before those with HIV. Future research must focus both on improving outcomes with currently available medications and rapidly evaluating the safety and efficacy of forthcoming antiviral compounds in HIV/HCV coinfected persons. PMID- 16356580 TI - Evaluation of a bioartificial liver based on a nonwoven fabric bioreactor with porcine hepatocytes in pigs. AB - BACKGROUND/AIMS: We developed a bioartificial liver (BAL) based on a direct hemoperfusion typed nonwoven fabric bioreactor containing porcine hepatocytes. In this study, the efficacy of our BAL was evaluated with a pig fulminant hepatic failure (FHF) model. METHODS: FHF was induced with intravenous administration of D-galactosamine (1.3 g/kg) in each pig. Twelve hours post D-galactosamine injection, fifteen pigs were divided into: a BAL group (n = 5), in which pigs received the BAL treatment with 1.0 to 1.3 x 10(9) hepatocytes for 6 h, a sham BAL group (n = 5), in which pigs received the BAL treatment without hepatocytes, and a FHF group (n = 5), in which pigs only received intensive care. Parameters related to liver function and animal survival up to 168 h were determined. RESULTS: In the BAL group, blood ammonia and plasma lactate levels were lower, and serum glucose levels and Fischer index were higher than those in the other two groups. Survival time of pigs in the BAL group was significantly prolonged as compared with the sham BAL and the FHF group. CONCLUSIONS: The BAL based on a nonwoven fabric bioreactor containing porcine hepatocytes appears to be effective in the treatment of FHF in pigs. PMID- 16356581 TI - Treatment options in HBV. AB - The available evidence on interferon-alpha (IFN) treatment for chronic hepatitis B is sufficient to conclude that in patients with HBeAg positive chronic hepatitis, standard IFN therapy significantly improves clearance of HBeAg (number needed to treat [NNT] = 4), loss of HBV-DNA (NNT = 4) and clearance of HBsAg (NNT = 18). HBeAg positive patients with normal or slightly raised ALT should be treated only if there is histological evidence of progressive disease. In patients with HBeAg negative chronic hepatitis, less than 20% of subjects who have achieved an end-of-treatment virological response after a course of standard IFN maintain a sustained virological response in the long-term. IFN treatment could help to delay or prevent disease decompensation and liver-related deaths but further large studies are needed. Lamivudine is effective at reducing, and sometimes clearing, HBV replication in heavily immunosuppressed patients and can be safely administered to patients with advanced liver disease. Lamivudine should be continued over an undefined extended period of time, with a switch from lamivudine to adefovir if there is an HBV-DNA breakthrough under therapy. Adefovir, excluding cost, is preferable to lamivudine as a first-choice because there is less chance of inducing resistance. The long-term benefit of lamivudine and adefovir and the role of combinations is under investigation. PMID- 16356582 TI - Management of hepatocellular carcinoma in human immunodeficiency virus-infected patients. AB - Hepatocellular carcinoma (HCC) resulting from chronic infection with hepatitis B or C virus (HBV, HCV) is a significant health problem. Concurrent infection with human immunodeficiency virus (HIV) may accelerate the progression from cirrhosis to HCC. Current guidelines advise screening patients with cirrhosis at 6-month intervals using ultrasonography and measurement of alpha-fetoprotein levels. In early-stage HCC, resection and liver transplantation are curative, as is percutaneous ethanol injection for small tumours in patients who are not candidates for surgery. HIV-infected patients do not qualify for liver transplantation. For late-stage HCC, chemoembolization can improve survival. Prevention of hepatitis and cirrhosis are key goals in reducing the impact of HCC. Numerous issues in HCC prevention, diagnosis, and management still remain to be resolved through large-scale, randomized clinical trials. PMID- 16356583 TI - Natural history and predictors of disease severity in chronic hepatitis C. AB - Cirrhosis is the end-stage consequence of fibrosis progression in patients with chronic hepatitis C. The median time from infection to cirrhosis is 30 years, with a high inter-individual variability, which is now better understood. Several factors have been clearly shown to be associated with fibrosis progression rate: duration of infection, age, male gender, alcohol consumption, HIV co-infection and low CD4 count. Metabolic conditions such as steatosis, being overweight and diabetes are emerging as independent co-factors of fibrogenesis. The recent validation of non-invasive biomarkers should facilitate the study of fibrosis progression in large populations. PMID- 16356584 TI - Retroposition of processed pseudogenes: the impact of RNA stability and translational control. AB - Human processed pseudogenes are copies of cellular RNAs reverse transcribed and inserted into the nuclear genome by the enzymatic machinery of L1 (LINE1) non-LTR retrotransposons. Although it is generally accepted that germline expression is crucial for the heritable retroposition of cellular mRNAs, little is known about the influences of RNA stability, mRNA quality control and compartmentalization of translation on the retroposition of processed pseudogenes. We found that frequently retroposed human mRNAs are derived from stable transcripts with translation-competent functional reading frames that are resistant to nonsense mediated RNA decay. They are preferentially translated on free cytoplasmic ribosomes and encode soluble proteins. Our results indicate that interactions between mRNAs and L1 proteins seem to occur at free cytoplasmic ribosomes. PMID- 16356585 TI - Molecular clocks: when times are a-changin'. AB - The molecular clock has proved to be extremely valuable in placing timescales on evolutionary events that would otherwise be difficult to date. However, debate has arisen about the considerable disparities between molecular and palaeontological or archaeological dates, and about the remarkably high mutation rates inferred in pedigree studies. We argue that these debates can be largely resolved by reference to the "time dependency of molecular rates", a recent hypothesis positing that short-term mutation rates and long-term substitution rates are related by a monotonic decline from the former to the latter. Accordingly, the extrapolation of rates across different timescales will result in invalid date estimates. We examine the impact of this hypothesis with respect to various fields, including human evolution, animal domestication and conservation genetics. We conclude that many studies involving recent divergence events will need to be reconsidered. PMID- 16356586 TI - Design and implementation of a smart card based healthcare information system. AB - Smart cards are used in information technologies as portable integrated devices with data storage and data processing capabilities. As in other fields, smart card use in health systems became popular due to their increased capacity and performance. Their efficient use with easy and fast data access facilities leads to implementation particularly widespread in security systems. In this paper, a smart card based healthcare information system is developed. The system uses smart card for personal identification and transfer of health data and provides data communication via a distributed protocol which is particularly developed for this study. Two smart card software modules are implemented that run on patient and healthcare professional smart cards, respectively. In addition to personal information, general health information about the patient is also loaded to patient smart card. Health care providers use their own smart cards to be authenticated on the system and to access data on patient cards. Encryption keys and digital signature keys stored on smart cards of the system are used for secure and authenticated data communication between clients and database servers over distributed object protocol. System is developed on Java platform by using object oriented architecture and design patterns. PMID- 16356587 TI - The use of a high-FeO olivine rock as a redox buffer in a nuclear waste repository. AB - Due to the higher stability of the spent nuclear fuel (mainly composed of UO2) under reducing conditions, and in order to enhance the retention/retardation of some key radionuclides, the olivine rock from the Lovasjarvi intrusion has been proposed as a potential redox-active backfill-additive in deep high-level nuclear waste (HLNW) repositories. In this work, two different approaches have been undertaken in order to establish the redox buffer capacity of olivine rock: (1) The capacity of the rock to respond to changes in pH or pe has been demonstrated and the final (pH, pe) coordinates agree with the control exerted by the system Fe(II)/Fe(III). (2) The rate of consumption of oxygen has been determined at different pH values. These rates are higher than the ones reported in the literature for other solids, what would point to the possibility of using this rock as an additive to the backfill material in a HLNW. PMID- 16356589 TI - In vitro biological activities of magainin alone or in combination with nisin. AB - Antimicrobial peptides have received increasing attention not only as potential candidates to their administration as antimicrobial agents, but also as potential drugs applied in cancer therapy. Here, we have examined the action of both nisin and magainin on human promyelocytic leukemia HL-60 cells. Cells were cultured in presence of either nisin or magainin 1 as well as in combination with both nisin and magainin 1. Results have revealed that magainin, but not nisin, produces a loss of cell viability in HL-60 cells, and a minor increase of hemolysis, whereas it is not responsible for cell membrane disruption and lactate dehydrogenase (LDH) leakage. In addition, magainin is involved in a significant generation of reactive oxygen species (ROS), as well as in an augment of caspase-3 activity. Magainin-induced apoptosis was verified by DNA fragmentation and annexin V FITC/propidium iodide (PI) staining of the cells. Promotion of cell death by magainin occurs via cytochrome c release accompanied by a substantial increase of proteasome activity. These results underline the importance of magainin as a drug capable of exerting an in vitro antitumoral activity by triggering apoptosis. PMID- 16356588 TI - AKAP350 modulates microtubule dynamics. AB - AKAP350 is a multiply spliced type II protein kinase A-anchoring protein that localizes to the centrosomes in most cells and the Golgi apparatus in epithelial cells. Multiple studies suggest that AKAP350 is involved in microtubule nucleation at the centrosome. Our previous studies demonstrated that AKAP350 was necessary for the maintenance of Golgi apparatus integrity. These data suggested that AKAP350 might be necessary for normal cytoskeletal interactions with the Golgi. To examine the relationship of AKAP350 with the microtubule cytoskeleton, we analyzed the effect of the depletion of AKAP350 on microtubule regrowth after nocodazole treatment in HeLa cells. The decrease in AKAP350 expression with short interfering RNA induced a delay in microtubule elongation with no effect on microtubule aster formation. In contrast, overexpression of the centrosomal targeting domain of AKAP350 elicited alterations in aster formation, but did not affect microtubule elongation. RNA interference for AKAP350 also induced an increase in cdc42 activity during microtubule regrowth. Our data suggest that AKAP350 has a role in the remodeling of the microtubule cytoskeleton. PMID- 16356590 TI - Design of genetically modified soybean proglycinin A1aB1b with multiple copies of bioactive peptide sequences. AB - The peptide IIAEK derived from beta-lactoglobulin has a hypocholesterolemic activity greater than that of beta-sitosterol. To create food proteins with multiple copies of this valuable peptide sequence, we introduced tandem multimers of the nucleotide sequence encoding the peptide into DNA regions corresponding to the five variable regions of soybean glycinin A1aB1b subunit, and expressed the mutants in Escherichia coli. The expression level and solubility of the five mutants, each containing four IIAEK sequences in each of the variable regions, were compared. Overall, the expression level and solubility of the mutants with four IIAEK sequences in the variable regions IV and V were the best followed by II > III > I. Further, introduction of the fifth IIAEK sequence to the variable region IV did not decrease expression level and solubility. Increasing the number of IIAEK to 7 and 10 slightly decreased expression level, while their solubility decreased to as low as 40 and 1%, respectively. Various mutations were combined to get a mutant containing as many IIAEK sequences as possible. Some of the resulting mutants were expressed in the soluble form. The mutant containing eight IIAEK from the combination of variable regions IV and V (IV-4 + V-4) showed the best balance of the expression level and solubility, followed by the combination of variable regions II and III (II-4 + III-4). The soluble fractions of these mutants were purified by hydrophobic, gel filtration and ion-exchange column chromatography. Yields of IIAEK peptide released by in vitro digestion with trypsin from both mutants were around 80%. This is the first report that a large amount of a physiologically active peptide could be introduced into food protein. PMID- 16356591 TI - Effects of central injection of angiotensin-converting-enzyme inhibitor and angiotensin type 1 receptor antagonist on the brain NF-kappaB and AP-1 activities of rats given LPS. AB - Angiotensin II (ANG II) activation of the angiotensin type 1 (AT1) receptor facilitates the production of brain interleukin-1beta (IL-1beta) and contributes to the induction of the fever following the intracerebroventricular (i.c.v.) injection of lipopolysaccharide (LPS). The purpose of the present study was to investigate whether proinflammatory transcription factors [nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1)] contribute to the ANG II-dependent production of cytokines within the brain. Interestingly, we found that a single i.c.v. injection of LPS had no effect on NF-kappaB and AP-1 activities in the hypothalamus, hippocampus, and cerebellum at either 1 or 3 h post-injection (except for a decrease in hypothalamic AP-1 activity at 1 h). Furthermore, both an angiotensin-converting-enzyme (ACE) inhibitor and an AT1 receptor antagonist enhanced (rather than reduced) the NF-kappaB and AP-1 activities in the hippocampus and/or cerebellum of rats given LPS. In contrast, an i.c.v. injection of ANG II increased the NF-kappaB activity in the hypothalamus. These results suggest that while "endogenous" ANG II exerts (via AT1 receptors) inhibitory effects on the activation of transcription factors in the brain of rats given LPS, a large dose of exogenous ANG II produces effects opposite to those induced by the presumably small amount of endogenous ANG II released locally by LPS. Our results seem not to support the idea that NF-kappaB and AP-1 play key roles in the ANG II-induced enhancement of the production of proinflammatory cytokines that is induced by LPS in the rat's brain. PMID- 16356592 TI - Immunohistochemical mapping of adrenomedullin in the human medulla oblongata. AB - We studied by immunocytochemistry the expression of adrenomedullin (AM) in the human medulla oblongata, sampled from 13 adult subjects (mean age: 38 years), whose medical history was negative for neurological and neurovascular pathologies. Immunoreactive neurons were found in the medulla oblongata with statistically significant differences among the various nuclei (one-way ANOVA, P < 0.001). The hypoglossal nucleus showed higher AM expression than that of the spinal tract of the trigeminal nerve (P < 0.05), solitary tract nucleus (P < 0.05), nucleus intercalatus (P < 0.05), and area postrema (P < 0.05). The arcuate nucleus and inferior olivary nuclear complex showed lower AM expression than the hypoglossal nucleus (P < 0.001), vestibular nuclei (P < 0.01), cuneate and gracile nuclei (P < 0.05), lateral column of the reticular formation (P < 0.05), and nucleus ambiguous (P < 0.05). Furthermore the nuclei were grouped with reference to their function, into somatic sensitive nuclei, somatic motor nuclei, visceral nuclei, reticular formation, and nuclei involved in cerebellar functions. The ANOVA revealed statistically significant differences (P < 0.001) in mean AM scores among the different groups. Nuclei involved in cerebellar function showed the lowest mean AM score (P < 0.05). The difference in AM score between somatic motor nuclei and visceral nuclei was also statistically significant (P < 0.05). Widespread AM immunoreactivity in the nuclei of the medulla oblongata may account for the role of the peptide in neuronal function and regulation of regional blood flow. Differences in the expression of AM in the nuclei studied indicate the different involvement of AM in neurotransmission and neuromodulation. PMID- 16356593 TI - Synthesis, antibacterial activity and QSAR studies of 1,2-disubstituted-6,7 dimethoxy-1,2,3,4-tetrahydroisoquinolines. AB - Some new substituted-tetrahydroisoquinoline derivatives were synthesized and evaluated for their in vitro antimicrobial activities against the standard Gram positive and Gram negative strains: Staphylococcus aureus (ATCC 25923), S. epidermidis (WHO-6), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), 1-aryl-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline 4a-c proved to be effective with MIC 3.5-20 (microg ml(-1)). Quantitative structure activity relationship (QSAR) studies with multiple linear regression analysis were applied to find correlation between different calculated molecular descriptors of the synthesized compounds and biological activity. PMID- 16356594 TI - Syntheses of new substituted triazino tetrahydroisoquinolines and beta-carbolines as novel antileishmanial agents. AB - A series of triazino tetrahydroisoquinolines (3-5) and beta-carboline derivatives (15-27) have been synthesized as novel antileishmanial agents. Among them, compounds 15, 16 and 25 have shown 78.0%, 78.6% and 68.0% in vivo inhibition against Leishmania donovani at a dose of 50 mg kg(-1) x 5 days, respectively, while compounds 3 and 18 exhibited 55.6% and 53.3% in vivo inhibitions, respectively, against L. donovani at a dose of 50 mg kg(-1) x 5 days. PMID- 16356595 TI - Comparison of tools for nutritional assessment and screening at hospital admission: a population study. AB - INTRODUCTION: This population study aimed to test the sensitivity and specificity of nutritional risk index (NRI), malnutrition universal screening tool (MUST) and nutritional risk screening tool 2002 (NRS-2002) compared to subjective global assessment (SGA) and to evaluate the association between nutritional risk determined by these screening tools and length of hospital stay (LOS). METHODS: Patients (n=995) were assessed at hospital admission by four screening tools (SGA, NRI, MUST and NRS-2002). Sensitivity, specificity and predictive values were calculated to evaluate NRI, MUST and NRS-2002 compared to SGA. Multiple logistic regressions, adjusted for age, were used to estimate odds ratios (OR) and confidence interval (CI) for medium and high, compared to low risk in patients hospitalized >11, compared to 1-10 days LOS. RESULTS: The sensitivity was 62%, 61% and 43% and specificity was 93%, 76% and 89% with the NRS-2002, MUST and NRI, respectively. NRS-2002 had higher positive (85%) and negative predictive values (79%) than the MUST (65% and 76%) or NRI (76% and 66%, respectively). Patients who were severely malnourished or at high nutritional risk by SGA (OR 2.4, CI 1.5-3.9), MUST (OR 3.1, CI 2.1-4.7) and NRS-2002 (OR 2.9, CI 1.7-4.9) were significantly more likely to be hospitalized >11 days, compared to 1-10 days, than patients assessed as low risk. CONCLUSION: NRS-2002 had higher sensitivity and specificity than the MUST and NRI, compared to SGA. There was a significant association between LOS and nutritional status and risk by SGA, NRS 2002, MUST and NRI. Nutritional status and risk can be assessed by SGA, NRS-2002 and MUST in patients at hospital admission. PMID- 16356597 TI - Immunogenicity of a single dose of reduced-antigen acellular pertussis vaccine in a non-vaccinated adolescent population. AB - German adolescents (n=123) without previous pertussis vaccination, no history of pertussis and low IgG-anti-pertussis-toxin (PT) levels received one dose of the Tdap vaccine Boostrix. Blood samples were taken before, and 5-12 days and 29-49 days after vaccination. IgG- and IgA-anti-PT, IgG- and IgA-anti filamentous hemagglutinin, IgG-anti-pertactin, IgG-anti-tetanus-toxin, and IgG-anti diphtheria-toxin were measured by ELISA. 88.6% of subjects had an immune response to PT, and all vaccinees had an immune response to at least one pertussis antigen 29-49 days after vaccination. IgA-anti-PT and IgA-anti-FHA responses were found in 43 and 81% of subjects, respectively. This study shows that in unvaccinated German adolescents pertussis immunity can be achieved by a single dose of Tdap. PMID- 16356596 TI - Chronic dehydration may impair renal function in patients with chronic intestinal failure on long-term parenteral nutrition. AB - BACKGROUND & AIMS: Renal impairment is a documented complication in long-term parenteral nutrition (LTPN) patients. However, the aetiologies have remained elusive. The aim of this study was to evaluate the impact of parenteral nutrition, digestive status, and hydration level on renal function in LTPN patients. METHODS: In a prospective study of 40 LTPN patients, renal function and hydration level were assessed by measurement of inulin and creatinine clearances, plasma creatinine, urea, aldosterone and renin and urinary sodium/potassium ratio. Patients were assigned to one of two groups according to their inulin clearance (normal = Group 1, 20% decrease or more = Group 2). RESULTS: Of the patients, 52.5% (21/40) had a decrease in glomerular filtration rate (-38 +/- 15%), with age taken into consideration. Patient characteristics, parenteral nutrition composition or duration and intestinal status were not different between the two groups. Urologic or nephrologic diseases were more frequent in Group 2 patients. Moreover, in Group 2 patients, a urinary sodium/potassium excretion ratio of less than 1 in 8/21 patients and plasma renin (316 +/- 298 vs. 86 +/- 53% of normal value) and aldosterone (291 +/- 464 vs. 58 +/- 36 pmol/l) that were significantly higher than in Group 1 patients suggested a hypovolemic component. CONCLUSION: Decreased renal function is frequent (52.5%) in LTPN patients. A volemic component was associated in more than 70% of them. An elevation of serum creatinine or an inversion of the urinary Na/K ratio requires an evaluation of hydration equilibration and an oral rehydration and a modification in parenteral nutrition formulation. PMID- 16356598 TI - Assessment of the compatibility of co-administered 7-valent pneumococcal conjugate, DTaP.IPV/PRP-T Hib and hepatitis B vaccines in infants 2-7 months of age. AB - This study assessed compatibility of concurrently administered 7-valent pneumococcal conjugate (PCV7), hepatitis B (HB) and DTaP.IPV/Hib vaccines. Infants were given DTaP.IPV/Hib and HB at 2, 4, 6 months and randomly assigned (2:1) to receive PCV7 concurrently or sequentially (at 3, 5, 7 months). Antibody levels were compared in 246 concurrent and 122 sequential vaccinees. Responses to PCV7, DTaP.IPV/Hib and HB were generally unaltered with concurrent administration except that Hib responses were increased (p=0.008) and HB responses were reduced (p=0.006) with concurrent dosing, the latter possibly from same thigh injection with DTaP.IPV/Hib. We conclude that PCV7, DTaP.IPV/Hib and HB are compatible with concurrent, separate injections. PMID- 16356599 TI - Interventions over 2 years to increase influenza vaccination of children aged 6 23 months in inner-city family health centers. AB - The purposes of this study were to test the ability of tailored interventions to raise influenza immunization rates and assess the effect on timely receipt of other vaccines. We conducted a before/after trial over 2 years to increase influenza vaccination rates of patients 6-23 months old in five inner-city family health centers serving low-income children with a sixth site as a concurrent control. Influenza vaccination rates improved significantly from a baseline of 4.7-24.7% in the first year and 36.6% in the second year, P < 0.001. The increase in rates was greater in intervention sites than the control site (31.9% versus 25.7%, P = 0.02). In regression analyses of influenza vaccination, intervention year was associated with an odds ratio (OR) of 9.4 (95% confidence interval (CI) = 4.4-20.0) for the first intervention year and OR = 13.4 (95% CI = 6.5-28.0) for the second intervention year. Children vaccinated against influenza were more likely to have received DTaP 3 and MMR within 2 months of the recommended age than children not vaccinated against influenza (P < 0.001). PMID- 16356600 TI - Immune response induced by Salmonella typhimurium defective in ppGpp synthesis. AB - Systemic infection by Salmonella typhimurium requires coordinated expression of virulence genes found primarily in Salmonella Pathogenecity Islands (SPIs). We have previously reported that the intracellular signal that induces these virulence genes is a stringent signal molecule, ppGpp [Song et al. J Biol Chem 2003;279:34183]. In this study, we found that relA and spoT double mutant Salmonella (DeltappGpp strain), which is defective in ppGpp synthesis, was virtually avirulent in BALB/c mice. Subsequently, the live vaccine potential of the avirulent DeltappGpp Salmonella strain was determined. A single immunization with live DeltappGpp Salmonella efficiently protected mice from challenge with wild-type Salmonella at a dose 10(6)-fold above the LD50 30 days after immunization. Various assays revealed that immunization of mice with the DeltappGpp strain elicited both systemic and mucosal antibody responses, in addition to cell-mediated immunity. PMID- 16356601 TI - Cost-effectiveness of universal pneumococcal vaccination for infants in Italy. PMID- 16356602 TI - Immunization by particle bombardment of antigen-loaded poly-(DL-lactide-co glycolide) microspheres in mice. AB - In the present study, we investigated whether poly-(DL-lactide-co-glycolide) (50:50) microspheres (PLG MS) containing a model antigen, ovalbumin (OVA), were delivered into mouse skin and the immune responses induced using a microparticulate bombardment system, Helios gene gun system, which can painlessly deliver the powdered drug through the stratum corneum to the epidermal-dermal interface using a high velocity supersonic flow of helium gas to accelerate the particles. The introduction of OVA-loaded PLG MS shows helium pressure dependence, so that improved introduction can be achieved by a higher helium pressure used, thereby inducing sufficient anti-OVA IgG level. Moreover, in order to determine the type of immune system induced using particle bombardment, we investigated helper T-cell response characterized by the cytokine production in the isolated splenocytes 6 weeks after immunization and consequent production of the anti-OVA IgG subclasses in the serum in mice. As a result, IL-4 production in splenocytes and anti-OVA IgG1 level were preferentially elicited by particle bombardment with OVA-loaded PLG MS compared with IFN-gamma and anti-OVA IgG2a level. It seemed likely that particle bombardment using this system led to a Th-2 type immune response, i.e. a humoral immune response. In conclusion, this microparticulate bombardment system is a promising immunization method, expected to become an alternative to needle injection used to administer a broad range of vaccines for the treatment of various diseases. PMID- 16356603 TI - Phase I safety and immunogenicity trial of FMP1/AS02A, a Plasmodium falciparum MSP-1 asexual blood stage vaccine. AB - We report the first safety and immunogenicity trial of the Plasmodium falciparum malaria blood stage vaccine candidate, FMP1/AS02A consisting of the FMP1 antigen, an Escherichia coli-expressed His-tagged fusion protein from the 42 kDa C terminal fragment from the 3D7 clone of the merozoite surface protein 1 formulated in the AS02A adjuvant. An open label, prospective, single-center Phase I dose escalation trial of FMP1/AS02A was conducted in 15 adult malaria-naive human volunteers to assess safety, reactogenicity, and immunogenicity. The vaccine was safe and well-tolerated and no serious adverse events were observed. The vaccine induced high-titer ELISA and IFA responses in all volunteers. Proliferative and ELISPOT responses were induced to vaccine antigen. Biologically active antibodies were induced as measured by GIA. This study establishes the foundation to further evaluate and measure the vaccine's ability to reduce morbidity and mortality in target populations directly affected by P. falciparum malaria. PMID- 16356604 TI - Radon exhalation of hardening concrete: monitoring cement hydration and prediction of radon concentration in construction site. AB - The unique properties of radon as a noble gas are used for monitoring cement hydration and microstructural transformations in cementitious system. It is found that the radon concentration curve for hydrating cement paste enclosed in the chamber increases from zero (more accurately - background) concentrations, similar to unhydrated cement. However, radon concentrations developed within 3 days in the test chamber containing cement paste were approximately 20 times higher than those of unhydrated cement. This fact proves the importance of microstructural transformations taking place in the process of cement hydration, in comparison with cement grain, which is a time-stable material. It is concluded that monitoring cement hydration by means of radon exhalation method makes it possible to distinguish between three main stages, which are readily seen in the time dependence of radon concentration: stage I (dormant period), stage II (setting and intensive microstructural transformations) and stage III (densification of the structure and drying). The information presented improves our understanding of the main physical mechanisms resulting in the characteristic behavior of radon exhalation in the course of cement hydration. The maximum value of radon exhalation rate observed, when cement sets, can reach 0.6 mBq kg(-1) s( 1) and sometimes exceeds 1.0 mBq kg(-1) s(-1). These values exceed significantly to those known before for cementitious materials. At the same time, the minimum ventilation rate accepted in the design practice (0.5 h(-1)), guarantees that the concentrations in most of the cases will not exceed the action level and that they are not of any radiological concern for construction workers employed in concreting in closed spaces. PMID- 16356605 TI - Expression of urokinase-type plasminogen activator receptor is correlated with metastases of lingual squamous cell carcinoma. AB - Lingual squamous cell carcinoma is common and the survival rate is relatively low. The invasion of cancer cells from the primary tumour into the surrounding tissue is an early step in the process of metastasis and urokinase-type plasminogen activator receptor (uPAR) is a vital mediator of cellular migration in some carcinomas. By binding urokinase-type plasminogen activator, uPAR localises proteolytic activity to the leading edge of the cells, thereby facilitating cellular migration and penetration through tissue boundaries. uPAR also binds directly to vitronectin and associates with integrins within the plasma membrane, which alters the strength of cellular adhesion. In this study we used reverse transcription polymerase chain reaction, immunocytochemistry, and Western-blot to examine the expression of uPAR mRNA and protein in Ts and Tca 8113 cell lines of lingual squamous cell carcinoma and in normal oral mucosal cells. uPAR mRNA and protein were expressed in Ts cells, but not in Tca 8113 cells or in normal oral epithelial cells. Ts cells have higher metastatic potential than Tca 8113 cells. The results suggest that uPAR has an important role in the aggressiveness of lingual squamous cell carcinoma. PMID- 16356606 TI - An easy and innovative technique for recording operation notes. PMID- 16356607 TI - Recurrent Kikuchi-Fujimoto disease: case report. AB - Kikuchi-Fujimoto disease, also known as histiocytic necrotising lymphadenitis, is a self-limiting condition of uncertain aetiology characterised by lymphadenopathy, pyrexia, and neutropenia. Some reported cases have been associated with systemic lupus erythematosus and there have been suggestions that Kikuchi's disease could represent a mild form of lupus but without definite evidence. We describe an unusual case of histiocytic necrotising lymphadenitis in an Asian woman who had recurrent episodes for five years before a diagnosis was made. PMID- 16356608 TI - A grounded theory study of Swedish women's experiences of inflammatory symptoms of the breast during breast feeding. AB - OBJECTIVE: to gain an understanding of the experiences of women who have had an episode of inflammatory symptoms of the breast during lactation. DESIGN: a grounded theory approach, inspired by Glaser and Strauss, was used in an interview study. SETTING: the study was carried out at a midwife-led breast feeding clinic in Sweden. PARTICIPANTS: the strategic sample consisted of 14 mothers, all of whom had had an episode of breast inflammation between 10 and 22 weeks before the interview. Nine were primiparous. Ages ranged between 24 and 38 years. FINDINGS: the analysis resulted in the emergence of one core category and five conceptual categories. The core category was 'the will to breast feed', and the five conceptual categories were 'perspectives on breast feeding', 'personal strategies', 'enduring and adjustment', 'support' and 'causal frameworks'. The core category was reflected in all the other categories and was dependent on them. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: an episode of breast inflammation is a possible threat to the maintenance of breast feeding, but the will to breast feed may make it possible for mothers to withstand physical and emotional difficulties caused by the illness. Breast-feeding mothers may have chosen a goal for the length of time they wish to breast feed, and the attainment of this goal is prestige-filled. Midwives, throughout the chain of care, could improve the amount and type of information given about the demands and possible complications of breast feeding. The availability of immediate clinical expertise for these mothers is an important factor, and health-care planners should be aware of this. PMID- 16356609 TI - An exploratory study in the UK of the effectiveness of three different pain management regimens for post-caesarean section women. AB - OBJECTIVE: To compare the effects of three types of analgesic administration after elective caesarean section on a number of clinical outcome measures. Supplementary aims of the study were to determine the acceptability of, and satisfaction with, the different regimens. DESIGN: A quasi-experimental different subject design was used to compare three types of analgesic administration on pain, post-operative nausea and vomiting, analgesic consumption, length of hospital stay and overall satisfaction with pain management. SETTING: A specialist women's hospital in a large UK city, with around 1500 caesarean sections per annum. PARTICIPANTS: 95 women who had undergone elective caesarean section. INTERVENTIONS: The women were allocated to one of the three pain management groups: group 1 (oral morphine, Co-dydramol and diclofenac [all self administered]); group 2 (oral morphine, Co-dydramol and diclofenac [all midwife administered]); and Group 3 (intra-muscular morphine, oral Co-dydramol and diclofenac [all midwife-administered]). The safety of self-medication was measured by adherence to a safety protocol. MEASUREMENTS: Data collection was undertaken over the first 3 days after surgery and included visual analogue scale (0-100mm) pain scores, analgesic consumption, incidence of post-operative nausea and vomiting, and length of hospital stay. In addition, questionnaires were given to midwives and patients to assess the acceptability of self-medication and patient satisfaction. Data collection took place between June 2002 and June 2003. FINDINGS: The results indicated that the outcomes of all three interventions were comparable in terms of pain scores, incidence of post-operative nausea and vomiting, and overall levels of satisfaction, although intra-muscular morphine was disliked to a degree that deterred some women from requesting it. Consumption of oral morphine was significantly greater than consumption of intramuscular injections of morphine, whereas Co-dydramol use was lower in the self-medicating group; the self-medicating women also went home, on average, a day earlier than women in the other two groups. PMID- 16356610 TI - A qualitative study exploring the support needs of first-time mothers on their journey towards intuitive parenting. AB - OBJECTIVE: to gain an understanding of the experiences of first-time mothers in the early weeks of motherhood in order to elicit what areas of support these women find empowering in easing their adjustment. DESIGN: a grounded theory approach was used. Data were collected through individual in-depth interviews. SETTING: an area in the South of England with maternity services provided by a consultant obstetric unit and four midwife-led centres. PARTICIPANTS: eight primiparous women aged 20-39 years, who had given birth normally at term to a healthy baby. FINDINGS: five categories 'expert to novice', 'losing touch', 'perceiving expertise', 'restoring balance' and 'falling into place' revealed a journey women travelled as they left behind their comfortable, controlled lives in which they were 'experts' and faced the unknown world of motherhood. Eventually, practice, support and knowledge shared with peers facilitated proficiency and intuitive mothering. Throughout this transition, the overriding concern of the mothers was to develop confidence and skills to give optimal care to their baby. 'Doing it right' emerged as the core category. IMPLICATIONS FOR PRACTICE: an understanding of the factors women considered to be supportive or inhibitive in easing their adjustment to motherhood might enable midwives to move beyond more traditional forms of postnatal care to explore innovative ways of providing and facilitating access to supportive resources. PMID- 16356611 TI - Squamous cell carcinoma of the lateral nail fold. AB - The digital nail complex is occasionally involved by squamous cell carcinoma. The published literature has either been indiscriminating of the site of origin of this tumour within the nail complex or has concentrated attention on the nail bed as the site of pathology. Tumours originating in the lateral nail fold can be clearly differentiated from those of the nail bed itself. This study identifies six cases of squamous cell carcinoma arising in the lateral nail fold. While surgical convention remains to amputate the digital tip for squamous cell carcinoma of any part of the nail complex, the dermatological literature identifies that local surgery can be curative for these tumours, when presenting early and without bone involvement, although offering no discussion of reconstruction. Reconstruction is desirable and methods of achieving this following local excision of lateral nail fold tumours are illustrated in this series. PMID- 16356612 TI - Relationship between CT intensity, micro-architecture and mechanical properties of porcine vertebral cancellous bone. AB - BACKGROUND: In vivo assessment of bone density is insufficient for the evaluation of osteoporosis in patients. A more complete diagnostic tool for the determination of bone quality is needed. Micro-computed tomography imaging allows a non-destructive method for evaluating cancellous bone micro-architecture. However, lengthened exposure to ionizing radiation prevents patients to be imaged by such a system. The aim for this study was to elucidate the relationships between image intensity (of Hounsfield units), cancellous bone micro-architecture and mechanical properties. METHODS: Using pig vertebral cancellous bone, the bone specimens were imaged using clinical and micro-computed tomography scanners and subsequently subjected to uniaxial compression testing. RESULTS: Results indicate that micro-architecture can be predicted using clinical image intensity. Micro architectural parameters relevant to osteoporosis study, such as percent bone volume, trabecular bone pattern factor, structure model index, trabecular thickness and trabecular separation have shown significant correlation with R2 values of 0.83, 0.80, 0.70, 0.72, and 0.54, respectively, when correlated to Hounsfield units. In addition, the correlation of mechanical properties (E, sigma yield, and sigma ult) in the superior-inferior direction (the primary loading direction), to micro-architecture parameters has also been good (R2 > 0.5) for all except tissue volume, tissue surface and degree of anisotropy. INTERPRETATION: This proves that the predictive power of bone strength and stiffness was improved with the combination of bone density and micro architecture information. This work supports the prediction of micro-architecture using current clinical computed tomography imaging technology. PMID- 16356613 TI - Effects of fusion-bone stiffness on the mechanical behavior of the lumbar spine after vertebral body replacement. AB - BACKGROUND: Implants for vertebral body replacement are often inserted together with an additional stabilizing implant, e.g. an internal fixation device. During implantation bone grafts or milled bone is normally added to the anterior implant. Little is known about the stiffening effect of this fusion-bone mass on the mechanical behavior of the corresponding bone region, including the load distribution between the different parts. METHODS: A three-dimensional finite element model of the lumbar spine was created with a vertebral body replacement at L3, a paired internal fixation device between L2 and L4, and left anterolateral fusion bone. The elastic modulus of fusion bone was varied in discrete steps between 0 MPa and 10,000 MPa. The model was loaded to simulate standing, 20 degrees flexion, 15 degrees extension and 6 degrees axial rotation in the lumbar spine. FINDINGS: The elastic modulus of fusion bone has a considerable effect on the compressive force on vertebral body replacement and fusion bone for all loading cases studied. For extension, it also affects intersegmental rotation, the force in the erector spinae muscle, the compressive force on the internal fixator and intradiscal pressure in the adjacent discs. The elastic modulus most strongly affects the different parameters at values between 0 MPa and 500 MPa. INTERPRETATION: Adding bone mass during vertebral body replacement reduces the loads on the ventral implant for all loading cases studied but extension when the fusion-bone stiffens. This protects the implant from fatigue. The load on the fusion bone increases with increasing elastic modulus. Thus bone grafts should be used whenever possible. PMID- 16356614 TI - What do we know about relapse in pathological gambling? AB - The study of pathological gambling and its treatment is in a nascent stage. To date, no consistent definition of relapse to gambling exists, and only a few empirical studies have evaluated the phenomenon of relapse directly in this patient population. Perspectives on relapse to drug and alcohol use appear relevant to the study of pathological gambling, and relapse to gambling is reviewed within the larger context of addictive disorders. The application of psychological, biological, environmental, and treatment factors are described as they may relate to relapse among pathological gamblers. PMID- 16356615 TI - A probabilistic effect assessment model for hazardous substances at the workplace. AB - A major problem in risk assessment is the quantification of uncertainties. A probabilistic model was developed to consider uncertainties in the effect assessment of hazardous substances at the workplace. Distributions for extrapolation factors (time extrapolation, inter- and intraspecies extrapolation) were determined on the basis of appropriate empirical data. Together with the distribution for the benchmark dose obtained from substance-specific dose response modelling for the exemplary substances 2,4,4-trimethylpentene (TMP) and aniline, they represent the input distributions for probabilistic modelling. These distributions were combined by Monte Carlo simulation. The resulting target distribution describes the probability that an aspired protection level for workers is achieved at a certain dose and the uncertainty associated with the assessment. In the case of aniline, substance-specific data on differences in susceptibility (between species; among humans due to genetic polymorphisms of N acetyltransferase) were integrated in the model. Medians of the obtained target distributions of the basic models for TMP and aniline, but not of the specific aniline model are similar to deterministically derived reference values. Differences of more than one order of magnitude between the medians and the 5th percentile of the target distributions indicate substantial uncertainty associated with the effect assessment of these substances. The probabilistic effect assessment model proves to be a practical tool to integrate quantitative information on uncertainty and variability in hazard characterisation. PMID- 16356616 TI - Making the invisible body visible. Bone scans, osteoporosis and women's bodily experiences. AB - The imaging technology of bone scans allows visualization of the bone structure, and determination of a numerical value. Both these are subjected to professional interpretation according to medical (epidemiological) evidence to estimate the individual's risk of fractures. But when bodily experience is challenged by a visual diagnosis, what effect does this have on an individual? The aim of this study was to explore women's bodily experiences after a bone scan and to analyse how the scan affects women's self-awareness, sense of bodily identity and integrity. We interviewed 16 Danish women (aged 61-63) who had had a bone scan for osteoporosis. The analysis was based on Merleau-Ponty's perspective of perception as an embodied experience in which bodily experience is understood to be the existential ground of culture and self. Women appeared to take the scan literally and planned their lives accordingly. They appeared to believe that the 'pictures' revealed some truth in themselves. The information supplied by the scan fostered a new body image. The women interpreted the scan result (a mark on a curve) to mean bodily fragility which they incorporated into their bodily perception. The embodiment of this new body image produced new symptom interpretations and preventive actions, including caution. The result of the bone scan and its cultural interpretation triggered a reconstruction of the body self as weak with reduced capacity. Women's interpretation of the bone scan reorganized their lived space and time, and their relations with others and themselves. Technological information about osteoporosis appeared to leave most affected women more uncertain and restricted rather than empowered. The findings raise some fundamental questions concerning the use of medical technology for the prevention of asymptomatic disorders. PMID- 16356617 TI - Effects of clozapine on memory function in the rat neonatal hippocampal lesion model of schizophrenia. AB - Clozapine is an effective atypical antipsychotic drug used to treat schizophrenia. It has the advantage of producing fewer extrapyramidal motor side effects than typical antipsychotic drugs such as haloperidol. Schizophrenia involves more than the hallmark symptom of psychosis. Substantial cognitive impairment is also seen. Effective drug treatments against the cognitive impairment of schizophrenia need to be developed. The current study was conducted to determine the effects of clozapine on working memory in the rat neonatal hippocampal lesion model of schizophrenia, which includes symptoms of cognitive impairment. Infant Sprague-Dawley rats were given ibotenic acid lesions of the hippocampus on day 7 of age (using the day of birth as day 0). Controls were given vehicle infusions. In adulthood, the rats were trained on the 8-arm radial maze using the win-shift procedure. After 6 sessions of training, the lesioned rats and their controls were administered repeated injections of saline or clozapine (2.5 mg/kg) for the next 12 sessions of training. The females had significant radial-arm maze choice accuracy impairments caused by either clozapine or the hippocampal lesion, but the combination of the two treatments had no additive effect. The males showed a different pattern of effects. Intact males did not show a significant clozapine-induced impairment, whereas males with hippocampal lesions did show significant clozapine-induced impairment although hippocampal lesions by themselves did not significantly impair male choice accuracy. These data show that clozapine can cause memory impairment and it potentiates rather than reverses hippocampal lesion-induced deficits. There are critical sex-related differences in these effects. PMID- 16356618 TI - 90-day oral (gavage) study in rats with galactooligosaccharides syrup. AB - A 90-day oral (gavage) study was conducted in male and female Sprague Dawley rats to investigate the safety of Vivinal galactooligosaccharides (GOS) syrup at 2500 or 5000 mg/kg bw/day. A reference control containing fructooligosaccharides (FOS) was used to match the oligosaccharide and digestible sugars in the test material (approximately 45% and 30%, respectively) and to assess if these had an impact on food consumption. Measurements included clinical observations, body weights, food consumption, hematology, clotting parameters, blood chemistries, urinalysis, ophthalmologic examinations, gross necropsies, organ weights, and histological examinations. There were no effects of feeding GOS syrup at either concentration on any parameter except food consumption. Statistically significant decreases (7 13%) in food consumption were seen in both sexes in the GOS syrup-treated animals at 5000 mg/kg bw/day and animals treated with the FOS control when compared to the reverse osmosis deionized (RODI) water controls. Based on the lack of toxicological effects in the study, the NOAEL for Vivinal GOS syrup is 5000 mg/kg bw/day when administered by gavage for 90 consecutive days. PMID- 16356619 TI - [Future research in immunology for composite tissue allotransplantation]. AB - Hand and composite tissue allotransplantation (CTA) holds great potential for reconstructive surgery but its development is currently limited by the side effects of the immunosuppressive drugs. Induction of specific tolerance, a situation where the recipient does not mount an immune response against the allograft but remains fully immunocompetent, holds exciting promise. Generation of mixed hematopoietic chimerism by infusing the recipient with donor bone marrow cells has been shown to induce tolerance without chronic immunosuppression. Genetic matching of the donor and the recipient is another option for transplanting composite tissues with only an initial course of immunosuppression. Experiments demonstrated long-term survival of musculoskeletal allografts between MHC-matched miniature swine. Finally, new immunosuppressive agents with a more targeted action will reduce side-effects and may prevent the development of chronic rejection. Skin-specific immunosuppression is particularly useful for limb transplants since skin, regarded as the most antigenic component, is easily accessible to topical or irradiation therapies. PMID- 16356620 TI - 2,3-Epithiopropyl methacrylate as functionalized monomer in a dental adhesive. AB - OBJECTIVES: The aim of this study was to investigate new dentin bonding agents formulated with 2,3-epithiopropyl methacrylate (ETMA) comonomer blends. METHODS: Synthesis and characterization of ETMA were performed. Adhesive resins with three different ETMA concentrations (0.1, 1 and 10 wt.%) were prepared. To comparison a blend with only Bis-GMA/HEMA and with 1% of GMA were used as controls. FTIR analysis was used to verify the degree of conversion. Blend copolymerization was investigated by (1)H NMR spectroscopy analysis. To investigate immediate properties of experimental adhesives, microtensile bond strength to human dentin and SEM fractographic analysis were studied. RESULTS: ETMA was synthesized with 90% yield through a one step synthetic route. Degree of conversion for the experimental and control resins was not significantly different. Different monomers showed copolymerization among them, not presenting traces of ETMA release in 0.1 and 1% groups. Bonding agents containing ETMA had statistically higher microtensile bond strength values than the no ETMA adhesive (p<0.05). CONCLUSIONS: Dental adhesive resin-ETMA mixtures produced increased bond strengths without harming the other properties investigated. PMID- 16356621 TI - Cholesterol effects on a mixed-chain phosphatidylcholine bilayer: a molecular dynamics simulation study. AB - A molecular dynamics simulation of a mono-cis-unsaturated 1-palmitoyl-2-oleoyl phosphatidylcholine bilayer containing approximately 22 mol% of cholesterol (POPC Chol) was carried out for 15 ns. An 8-ns trajectory was analysed to determine the effects of Chol on the membrane properties and compare it with that on the fully saturated 1,2-dimyristoyl-phosphatidylcholine bilayer containing approximately 22 mol% of Chol (DMPC-Chol). The study suggests that the experimentally observed weaker effect of Chol on the POPC than DMPC bilayer might result from a different vertical localisation of the Chol hydroxyl group (OH-Chol) in both bilayers: in the POPC-Chol bilayer, OH-Chol is placed approximately 3 A higher in the bilayer interface than in the DMPC-Chol bilayer. Because of the rigid cis double bond in the beta-chain of POPC, Chol fits worse to the POPC-Chol membrane environment and is pushed up, in effect all Chol ring atoms are, on average, located above the double bond. Both in mono-cis-unsaturated and fully saturated PC bilayers, Chol induces stronger van der Waals interactions among the chains, whereas its interactions with the chains are weak. In contrast to DMPC, the smooth alpha-face of the Chol ring lowers the order of POPC chains, whereas the rough beta-face increases the order. PMID- 16356622 TI - Fos, nociception and the dorsal horn. AB - The protooncogene c-fos is rapidly activated after noxious stimuli to express the protein Fos in spinal dorsal horn neurons that are in the 'correct' locations for nociceptive information transfer. As such, therefore, mapping Fos expression in these neurons is at present the best global marker for efficiently locating populations of neurons in the awake animal that respond to nociceptive input. This allows, among other things, precise behavioral measurements to be correlated with Fos expression. Two arenas where mapping dorsal horn Fos expression has made a major impact are in the anatomy of nociceptive systems and as a useful assay for the analgesic properties of various therapeutic regimens. Also Fos expression is the only way to map populations of neurons that are responding to non localized input such as withdrawal after addiction and vascular occlusion. Another insight is that it shows a clear activation of neurons in superficial 'pain-processing' laminae by innocuous stimuli after nerve lesions, a finding that presumably bears on the allodynia that often accompanies these lesions. It is to be understood, however, that the Fos localizations are not sufficient unto themselves, but the major function of these studies is to efficiently locate populations of cells in nociceptive pathways so that powerful anatomic and physiologic techniques can be brought to bear efficiently. Thus, the purpose of this review is to summarize the studies whose numbers are geometrically expanding that deal with Fos in the dorsal horn and the conclusions therefrom. PMID- 16356623 TI - Management of a Wilms' tumour, in a Jehovah's Witness, 30+ weeks pregnant. PMID- 16356624 TI - Existence of lipid microdomains in bilayer of dipalmitoyl phosphatidylcholine (DPPC) and 1-stearoyl-2-docosahexenoyl phosphatidylserine (SDPS) and their perturbation by chlorpromazine: a 13C and 31P solid-state NMR study. AB - The polyunsaturated fatty acid docosahexaenoic acid (DHA, 22:6, n-3) is found at a level of about 50% in the phospholipids of neuronal tissue membranes and appears to be crucial to human health. Dipalmitoyl phosphatidylcholine (DPPC, 16:0/16:0 PC) and the DHA containing 1-stearoyl-2-docosahexenoyl phosphatidylserine (SDPS) were used to make DPPC (60%)/SDPS (40%) bilayers with and without 10 mol% chlorpromazine (CPZ), a cationic, amphiphilic phenothiazine. Resonances that are present in 13C NMR spectrum of the DPPC (60%)/SDPS (40%) sample and that disappear in presence of 10% CPZ most probably are due to the special interface environment, e.g. the hydrophobic mismatch, at the interface of DPPC and SDPS microdomains in the DPPC/SDPS bilayer. In itself the appearance of resonances at novel chemical shift values is a clear demonstration of a unique chemical environment in the DPPC (60%)/SDPS (40%) bilayer. The findings of the study presented here suggest CPZ bound to the phosphate of SDPS will slow down and partially inhibit such a DHA acyl chain movement in the DPPC/SDPS bilayer. This would affect the area occupied by a SDPS molecule (in the bilayer) and probably the thickness of the bilayer where SDPS molecules reside as well. It is quite likely that such CPZ caused changes can affect the function of proteins embedded in the bilayer. PMID- 16356625 TI - Concentration of potentially toxic elements in field crops grown near and far from cities of the Pampas (Argentina). AB - Soils of the Pampas show no signs of contamination with potentially toxic elements (PTEs), except in the areas that surround cities. The concentration of PTE in crops grown in this region is in most cases unknown. Our objective was to determine the PTE concentration in main field crops and pastures grown in 'a priori' non-contaminated areas and in potentially contaminated areas around cities. Forty-eight plots from farms located far from cities or roads and 36 plots from farms located very close to cities or to high traffic roads were sampled. The area ranged from 33 degrees 40' S to 36 degrees 0' S and from 57 degrees 35' W to 61 degrees 22' W. Maize, soybean, wheat, grazed grassland and pastures, and their top soils were sampled. All samples were acid digested. Cadmium, zinc, chromium, copper, nickel and lead were determined using ICPES. Standard t-tests were performed. All soils were within the known normal values of soil PTE concentrations, with the exception of a few cases around cities. PTE accumulation on grains and aerial biomass is considered almost negligible in crops grown in both studied areas, with the exception of a few elements in soybean. PTE concentrations in crops and pastures are in keeping with the low content of trace metals found in soils of the Pampas. PMID- 16356626 TI - Information processing in brain microtubules. AB - Models of the mind are based on the idea that neuron microtubules can perform computation. From this point of view, information processing is the fundamental issue for understanding the brain mechanisms that produce consciousness. The cytoskeleton polymers could store and process information through their dynamic coupling mediated by mechanical energy. We analyze the problem of information transfer and storage in brain microtubules, considering them as a communication channel. We discuss the implications of assuming that consciousness is generated by the subneuronal process. PMID- 16356627 TI - Regulation of thyroid hormone receptor alpha2 RNA binding and subcellular localization by phosphorylation. AB - Thyroid hormone receptor alpha2 (TRalpha2) is an alternative splice product of the TRalpha primary transcript whose unique carboxyl terminus does not bind T3 or activate transcription. The physiological function of TRalpha2 is unknown. We have found that TRalpha2 is a single stranded RNA binding protein and that the RNA binding domain localizes to a 41 amino acid region immediately distal to the second zinc finger. TRalpha2 contains a single protein kinase CK2 phosphorylation site in its amino terminus and potentially nine CK2 sites in its unique carboxyl terminus. In vitro CK2 treatment of TRalpha2 eliminated its RNA binding. Mutational analysis indicated that phosphorylations at the N- and C-terminal sites both contribute to this inhibitory effect. Cellular localization studies demonstrated that phosphorylated TRalpha2 is primarily cytoplasmic, whereas unphosphorylated TRalpha2 is primarily nuclear. Since RNA binding is a property of unphosphorylated TRalpha2, the TRalpha2-RNA interaction likely represents a nuclear function of TRalpha2. PMID- 16356628 TI - Steroidogenesis in zebrafish and mouse models. AB - Steroid hormones regulate physiological homeostasis for salt, sugar, and sex differentiation. All steroids are synthesized from a common precursor, cholesterol, in a step that converts cholesterol to pregnenolone. The enzyme carrying out this first conversion step is CYP11A1. To further investigate the importance of steroid biosynthesis, animal models with defects in the Cyp11a1 gene are used. Mice with targeted disruption of the Cyp11a1 gene produce no steroids with severe adrenal defects. These mice survive during embryogenesis, but die after birth. Zebrafish with a block in cyp11a1 gene function has an earlier defect, presumably because it lacks adequate maternal steroid supply. When cyp11a1 activity was compensated by the injection of antisense morpholino oligos, the embryos have shortened axis and a defect of epibolic cell movement during early embryogenesis. The discovery of steroid function in cell movement is novel, and should provide new insights into our understanding of diverse functions of steroids. PMID- 16356630 TI - Analysis of the 5' flanking regions of human and murine HSD17B7: identification of a cholesterol dependent enhancer region. AB - 17Beta hydroxysteroid dehydrogenase type 7 (HSD17B7) was described to possess dual functionality in steroidogenesis as well as in postsqualene cholesterol biosynthesis in vitro. In order to gain insight into the transcriptional regulation, and thereby into in vivo functionality of HSD17B7, we analyzed and compared the 5' flanking regions of the corresponding human and murine genes. For this task we used bioinformatic and experimental approaches. The identified proximal promoter regions of both human and murine HSD17B7 genes contain multiple transcription factor binding sites and show strong similarity to cholesterogenic genes, especially to other postsqualene genes, but not to other steroidogenic genes. In liver cell lines, the transcriptional activity is dependent on the level of cholesterol, but not estradiol. The results of our study lead us to the conclusion that HSD17B7 is involved in postsqualene cholesterol biosynthesis in both human and mice. PMID- 16356629 TI - Dominant negative FTase (DNFTalpha) inhibits ERK5, MEF2C and CREB activation in adipogenesis. AB - We recently demonstrated that dominant negative FTase/GGTase I alpha-subunit inhibited (DNFTalpha-inhibited) insulin-stimulated adipocytes differentiation. DNFTalpha interferes with Ras prenylation whereby ERK1/2, CREB and the differentiation cascade are downregulated. To further investigate prenylation in adipogenesis, we examined DNFTalpha's ability to inhibit activation of ERK5, MEF2C and CREB. DNFTalpha-inhibited insulin-stimulated expression, activation and nuclear translocation of ERK5. Inhibition was associated with decreased activation of MEF2C and CREB by 80 and 78%, respectively. PD98059 did not block activation of ERK5 and MEF2C, but inhibited CREB phosphorylation by 90%. ERK5 siRNA-inhibited MEF2C activation, whereas it reduced CREB phosphorylation only 50%. Pre-adipocytes expressing DNFTalpha or treated with PD98059 were unable to differentiate to mature adipocytes, whereas pre-adipocytes transfected with ERK5 siRNA showed moderate inhibition of insulin-induced adipogenesis. Taken together, these data suggest that prenylation plays a critical role in insulin-stimulated adipogenesis, and that the ERK5 plays an important, but less crucial role in adipogenesis as compared to ERK1/2. PMID- 16356631 TI - Time-varying spreading depression waves in rat cortex revealed by optical intrinsic signal imaging. AB - This study aimed to investigate the variation of propagation patterns of successive spreading depression (SD) waves induced by K+ in rat cortex. SD was elicited by 1 M KCl solution in the frontal cortex of 18 Sprague-Dawley rats under alpha-chloralose/urethane anesthesia. We applied optical intrinsic signal imaging (OISI) at an isosbestic point of hemoglobin (550 nm) to examine regional cerebral blood volume (CBV) changes in the parieto-occipital cortex. In 6 of the 18 rats, OISI was performed in conjunction with DC potential recording of the cortex. CBV changes appeared as repetitive propagation of wave-like hyperemia at a speed of 3.7+/-0.4 mm/min, which was characterized by a significant negative peak (-14.3+/-3.2%) in the reflectance signal. Among the observed 186 SDs, the first wave always propagated through the entire imaged cortex in every rat, whereas following waves were likely to bypass the medial area of the imaged cortex (partially propagated waves, n=65, 35%). Correspondingly, DC potential shifts showed non-uniform in the medial area, and they seemed closely related to the changes in reflectance. For partially propagated SD waves, the mean time interval to the previous SD wave (217.0+/-24.3 s) was significantly shorter than for fully propagated SD waves (251.2+/-29.0 s). The results suggest that the propagation patterns of a series of SD waves are time-varying in different regions of rat cortex, and the variation is related to the interval between SD waves. PMID- 16356632 TI - alpha2-Adrenoceptor-mediated modulation of the release of GABA and noradrenaline in the rat substantia nigra pars reticulata. AB - The control of movement by the basal ganglia is influenced by inputs from diverse brain structures. Unfortunately, the mechanisms of modulation are poorly defined. Based on neuroanatomical evidence for alpha2A and alpha2C subtypes of alpha2 adrenergic receptors within this region, we hypothesize that noradrenergic alpha2 receptors can influence transmitter release in the SNr. To test this hypothesis we examined the effect of the alpha 2 adrenergic agonist, clonidine, and antagonist, rauwolscine, on the efflux of [3H]-GABA and [3H]-noradrenaline from brain slices of the rat substantia nigra pars reticulata. At low concentrations (10 nM), rauwolscine caused an 84.2 +/- 18.51% (p < 0.01) increase in KCl-evoked GABA release. At higher concentrations, rauwolscine caused a dose-dependent return to basal levels. Rauwolscine also enhanced basal GABA efflux after KCl washout with a similar biphasic concentration-dependence. Surprisingly, clonidine also enhanced [3H]-GABA release but had no effect on KCl-evoked [3H]-GABA release at concentrations which inhibited [3H]-NA efflux. These effects were potentiated by the GABA re-uptake inhibitor nipecotic acid. Together, our data indicate an important role for noradrenergic modulation in the SNr. The enhancing effect of both the alpha2 adrenoceptor agonist and antagonist on GABA release, while appearing paradoxical, can be rationalised by actions at distinct subsets of alpha2 adrenoceptors, using a simple model where alpha2A adrenoceptors are localized on the terminals of noradrenergic afferents impinging upon alpha2C adrenoceptor-containing GABAergic striato-nigral neurones. PMID- 16356633 TI - HIV-1 Tat neurotoxicity in primary cultures of rat midbrain fetal neurons: changes in dopamine transporter binding and immunoreactivity. AB - HIV-1 neurotoxic proteins (Tat, gp120) are believed to play a major role in pathogenesis of dementia in a significant portion of the AIDS patient population. Dopaminergic systems appear to be particularly important in HIV-associated dementia. In the current studies, we determined that primary cell cultures prepared from the midbrain of 18-day-old rat fetuses are sensitive to Tat neurotoxicity and investigated the possible effects of Tat on DAT-specific ligand binding and DAT immunoreactivity in rat fetal midbrain cultures. We found that Tat neurotoxicity was associated with a significant decrease in [3H]WIN 35428 binding. Immunostaining of cell cultures with antibodies recognizing the C-end epitope of DAT did not reveal significant changes in DAT immunoreactivity. The results of this study implicate involvement of monoamine transmission systems in HIV-associated dementia. PMID- 16356634 TI - Visuotactile temporal order judgments in ageing. AB - We report an experiment on the effects of ageing on crossmodal temporal perception. Young (mean age = 21.7 years) and old (mean age = 75.1 years) participants were presented with pairs of visual and vibrotactile stimuli to either hand and required to make unspeeded temporal order judgments (TOJs) regarding which sensory modality appeared to have been presented first. The stimulus onset asynchrony (SOA) between the two stimuli was varied using the method of constant stimuli. Temporal precision, as indexed by the just noticeable difference (JND), was better (i.e., JNDs were lower) when the stimuli were presented from different positions (M = 101 ms) rather than from the same position (M = 120 ms), as has been demonstrated previously. Additionally, older observers required more time (i.e., their JNDs were larger) to accurately perceive the temporal order (M = 131 ms) as compared to younger observers (M = 98 ms). Our results confirm that ageing deleteriously affects crossmodal temporal processing even when the spatial confound inherent in previous research has been ruled out. PMID- 16356635 TI - Attenuation of amyloid beta (Abeta)-induced inhibition of phosphatidylinositol 4 kinase activity by Abeta fragments, Abeta20-29 and Abeta31-35. AB - We previously reported that pathophysiological concentrations of amyloid beta protein (Abeta25-35, 0.1-10 nM) directly inhibited type II phosphatidylinositol 4 kinase (PI4KII) activity in neuronal plasma membranes, which resulted in the enhanced glutamate neurotoxicity. In the present study, we examined the effects of Abeta fragments, Abeta20-29 and Abeta31-35, on the 10 nM Abeta25-35- or Abeta1 42-induced inhibition of PI4KII activity. Both of the peptide fragments recovered the inhibition of rat brain plasma membrane PI4KII activity over the concentration range of 0.1-5 nM. Such protection by the Abeta fragments was observed in the 10 nM Abeta25-35-induced inhibition of recombinant human PI4KII, suggesting that these Abeta fragments blocked the inhibition on PI4KII molecule. The Abeta25-35-induced enhancement of glutamate neurotoxicity was also completely inhibited in the presence of these fragments. Thus, Abeta20-29 and Abeta31-35 ameliorated the Abeta-enhanced glutamate neurotoxicity probably through attenuation of Abeta-induced inhibition of PI4KII activity. PMID- 16356636 TI - Early increase in mRNA levels of pro-inflammatory cytokines and their interactions in the mouse hippocampus after transient global ischemia. AB - There is convincing evidence that cytokines are involved in the inflammatory response following cerebral ischemia, but the interactions among the pro inflammatory cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 in the early stage of ischemic reperfusion are not yet completely understood. In this study, we examined the early mRNA expressions of pro inflammatory cytokines in the ischemic hippocampus after 30 min of bilateral common carotid artery occlusion in C57BL/6J wild-type (WT) and TNF-alpha, IL 1alpha/beta or IL-6 gene knockout (KO) mice utilizing real-time polymerase chain reaction. The mRNA expressions of the pro-inflammatory cytokines TNF-alpha, IL-6 and IL-1beta were significantly induced in ischemic WT mice compared with in the sham-operated mice. These increases peaked at 3 to 24 h for TNF-alpha, at 12 h for IL-1beta, and at 6 to 24 h for IL-6 after ischemia. The pattern of temporal expression of the cytokine mRNAs in ischemic gene KO mice, however, differed from that in WT mice. The TNF-alpha mRNA expression showed a similar temporal expression pattern in IL-6 KO mice compared to in WT mice following ischemic reperfusion, and the levels at all time points were lower than in WT mice. The IL 1beta mRNA level was very low in ischemic TNF-alpha KO mice and IL-6 KO mice in spite of a small peak observed in both at 24 h. The IL-6 mRNA level was significantly upregulated at all time points in both ischemic WT and TNF-alpha KO mice; however, the peak was delayed by 12-h in IL-1alpha/beta KO mice. In conclusion, the present study indicates that the rapid increases in cytokine levels are interdependent, interactive, and possibly modulate each other in the mouse hippocampus after transient global ischemia. PMID- 16356637 TI - Examining task-dependencies of different attentional processes as reflected in the P3a and reorienting negativity components of the human event-related brain potential. AB - Unexpected changes in task-irrelevant auditory stimuli are capable to distract processing of task-relevant visual information. This effect is accompanied by the elicitation of event-related potential (ERP) components associated with attentional orientation, i.e. P3a and reorienting negativity (RON). In the present study we varied the demands of a visual task in order to test whether the RON component -- as an index of attentional reorientation after distraction -- is confined to a semantic task requiring working memory. In two ERP experiments we applied an auditory-visual distraction paradigm in which subjects were instructed to discriminate visual stimuli preceded by a task-irrelevant sound, this being either a standard tone (600 Hz, 88%) or a deviant tone (660 Hz, 12%). The visual stimuli were numbers which had to be judged on basis of a semantic (odd or even) or physical feature (either size or colour). As expected, deviance related ERP components namely the mismatch negativity (MMN), P3a, and RON were elicited. Importantly, the RON was affected by the variation of the task: within the semantic task an early RON and within the physical task a late RON was obtained. These results suggest that the RON component reflects two functionally distinct processes of attentional allocation after distraction: refocusing on task relevant information on the working memory level, and general reorientation of attention, e.g. preparation for the upcoming task. PMID- 16356638 TI - Rapid eye movement sleep deprivation induces changes in the high-affinity binding of [3H]-ouabain to the rat cortical membranes. AB - Rapid eye movement sleep (REMS) suppresses seizures. On the other hand, REMS deprivation (REMSD) increases brain susceptibility to seizures. Sodium potassium/ATPase is involved in the control of brain excitability. Ouabain, a cardiotonic glycoside, binds to a regulatory extracellular allosteric site in the sodium-potassium/ATPase inhibiting/stimulating its activity depending on its concentration. Endogenous ouabain-like substances exist in the brain; therefore, changes in the ouabain binding site may be involved in the increased brain excitability induced by REMSD. Adult, Wistar male rats were deprived of REMS for 96 hours by the flower-pot method (REMSD). A stress control group was kept in the same environment on a larger platform (LP). A third group of rats was kept in the same room in their home-cages (CONTROL). After REMSD all rats were sacrificed by decapitation and their cerebral cortex dissected. High-affinity [3H]-ouabain binding was carried out in cortical crude membrane preparation using 8 concentrations of [3H]-ouabain (1-24 nM). The results show a statistically significant increase of KD in the REMSD rats compared to both CONTROL and LP groups. There were no statistically significant differences in the Bmax among the experimental groups. There was also no change either in cortical activity of K+ stimulated p-nitrophenylphosphatase, the dephosphorylation reaction of phosphorylated sodium-potassium/ATPase or in Mg2+-stimulated p nitrophenylphosphatase. An increase in the KD of [3H]-ouabain binding to the sodium-potassium/ATPase in REMSD rats indicates a lower affinity to the endogenous inhibitors/stimulators of the enzyme. Therefore, this decreased affinity of the endogenous ouabain-like substances may be involved in the increased excitability induced by REMSD. PMID- 16356639 TI - Morphine protects for head trauma induced cognitive deficits in mice. AB - Victims of minor traumatic brain injury (mTBI) can show long lasting cognitive, emotional and concentration difficulties, amnesia, depression, apathy and anxiety. The symptoms are generally known as a post-concussive syndrome without clear morphological brain defects. Endogenous opiates are released after impact to the brain, suggesting they may play a role in TBI pathophysiology. Furthermore, the administration of opiates to the brain of injured animals has been shown to affect the injury, induce cellular changes and also have protective qualities for neurological impairments. Here, we examined the protective properties of the opiate morphine on cognitive performances following minimal brain injury in mice. For this purpose, we have used our non-invasive closed-head weight drop model in mice, which closely mimics real life mTBI and examined mice performance in the Morris water maze. Our procedure did not cause visible structural or neurological damage to the mice. A single morphine injection administrated immediately after the induction of minimal TBI protected the injured mice from cognitive impairment, checked 30, 60 and 90 days post injury. However, mice injected with morphine that were examined 7 days after the injury did not show better performance than the saline injected mice. Our results indicate that morphine has long but not short-term effects on the cognitive ability of brain-injured mice. Although the exact nature of opioid neuroprotection is still unknown, its elucidation may lead to the much-needed treatment for traumatic brain injury. PMID- 16356640 TI - The 5-HT- and FMRFa-activated signaling pathways interact at the level of the Erk MAPK cascade: potential inhibitory constraints on memory formation. AB - The sensorimotor synapse of Aplysia exhibits long-term facilitation (LTF) and long-term depression (LTD) elicited by the neuromodulator serotonin (5-HT) and the peptide Phe-Met-Arg-Phe-NH(2), respectively. 5-HT-induced LTF engages extracellular-regulated kinase (Erk) and CREB1, whereas FMRFa-induced LTD engages p38 MAPK (mitogen-activated protein kinase) and CREB2. The interaction of the 5 HT and FMRFa pathways was recently investigated in Aplysia at the level of gene expression. However, little is known about crosstalk of these pathways at the level of the second messenger cascades. We investigated the potential interaction of the 5-HT and FMRFa pathways at the level of the Erk cascade. We found that FMRFa inhibited basal Erk activity through p38 MAPK. FMRFa also inhibited 5-HT induced phosphorylation of Erk and nuclear accumulation of phospho-ERK, suggesting that FMRFa may place inhibitory constraints on memory formation through regulation of the Erk MAPK cascade. PMID- 16356641 TI - Expression of the cannabinoid CB2 receptor in the rat cerebellum: an immunohistochemical study. AB - Reports of cannabinoid CB2 receptor protein in the brain have been ambiguous. We therefore tested for CB2 immunoreactivity in the rat brain using immunofluorescence. We detected CB2 labeling in fine fibers in the granule layer. This CB2 labeling did not co-localise with the astrocyte marker glial fibrillary acidic protein (GFAP) and, therefore, the CB2-positive fibers were not astrocytes and were possibly microglial or neuronal. Additionally, strong CB2 labeling was detected in capillary endothelia in the granule, Purkinje cell, and molecular layers. Our results suggest that the role of CB2 receptors in the brain may have been previously underestimated. PMID- 16356642 TI - Dietary restriction increases hippocampal neurogenesis by molecular mechanisms independent of NMDA receptors. AB - Neurogenesis occurs in the adult hippocampus and is promoted by various stimulatory factors, such as enriched environments, running exercises, learning and dietary restriction. We have previously shown that the promotion of adult neurogenesis in the hippocampus by running wheel exercise is mediated by NMDA receptors. In the present study, we report that dietary restriction enhanced the hippocampal neurogenesis in mice lacking NMDA receptor epsilon1 subunit as well as wild-type mice. This result suggests that the molecular mechanism by which dietary restriction promotes adult neurogenesis may be, in contrast to that of running exercise, independent on the NMDA receptor functions. PMID- 16356643 TI - No association of the Val66Met polymorphism of brain-derived neurotrophic factor (BDNF) to multiple sclerosis. AB - Brain-derived neurotrophic factor (BDNF), a neurotrophin produced by neurons and immune cells, promotes neuronal survival and repair during development and after CNS injury. The BDNF-Val66Met polymorphism is functional and induces abnormal intracellular trafficking and decreased BDNF release. Therefore, we investigated the impact of the BDNF-Val66Met polymorphism on the susceptibility and clinical course in a case-control study of 224 multiple sclerosis (MS) Spanish patients and 177 healthy controls. We found no evidence for association to susceptibility or severity of the disease in our population. Moreover, we did not observe, in a subgroup of 12 MS patients, that the methionine substitution at position 66 in the prodomain had negative impact in the capacity to produce BDNF by peripheral blood mononuclear cells (PBMC). PMID- 16356644 TI - Family-based and case-control association studies of glutamate receptor GRIK3 Ser310Ala polymorphism in Polish patients and families with alcohol dependence. AB - The aim of this study was to evaluate the role of the GRIK3 functional polymorphism (Ser310Ala) in the pathogenesis of alcoholism. This polymorphism was investigated in two types of studies: (1) the association study in a whole group of alcoholics (116 patients fulfilling ICD-10 alcohol dependence (AD) criteria and 255 controls, Polish descent) and homogenous overlapping subgroups of patients with: a history of delirium tremens and/or alcohol seizures, early age of onset of alcoholism (AOO<26 years), a co-occurrence of dissocial personality disorder, a history of familial alcoholism; (2) the family-based study (using Transmission Disequilibrium Test (TDT) in 100 Polish families with alcohol dependence). The history of alcoholism was obtained using SSAGA (Polish version). GRIK3 functional polymorphism was determined using PCR. TDT revealed an adequate transmission of both alleles to the affected offspring in the whole group of alcohol families (29 x Ser, 24 x Ala; chi2=0.472; d.f.=1; p=0.492) and in the homogenous subgroups of families. No significant associations between any of the above mentioned alcohol phenotypes and Ser310 allele were observed (the whole AD group: p=0.66 AD with delirium and/or seizures: p=0.521; early onset AD: p=0.868; AD with familial history of alcoholism: p=0.798 and AD with dissocial personality disorder: p=0.618). These findings do not seem to support the hypothesis of the role of this polymorphism in the pathogenesis of alcoholism. PMID- 16356645 TI - Quantification of vesicles in differentiating human SH-SY5Y neuroblastoma cells by automated image analysis. AB - A new automated image analysis method for quantification of fluorescent dots is presented. This method facilitates counting the number of fluorescent puncta in specific locations of individual cells and also enables estimation of the number of cells by detecting the labeled nuclei. The method is here used for counting the AM1-43 labeled fluorescent puncta in human SH-SY5Y neuroblastoma cells induced to differentiate with all-trans retinoic acid (RA), and further stimulated with high potassium (K+) containing solution. The automated quantification results correlate well with the results obtained manually through visual inspection. The manual method has the disadvantage of being slow, labor intensive, and subjective, and the results may not be reproducible even in the intra-observer case. The automated method, however, has the advantage of allowing fast quantification with explicitly defined methods, with no user intervention. This ensures objectivity of the quantification. In addition to the number of fluorescent dots, further development of the method allows its use for quantification of several other parameters, such as intensity, size, and shape of the puncta, that are difficult to quantify manually. PMID- 16356646 TI - Emergence of endoplasmic reticulum stress and activated microglia in Purkinje cell degeneration mice. AB - In the current studies, we characterized the molecular and cellular mechanism of cell death in Purkinje cell degeneration (pcd) mice using real-time quantitative PCR, immunohistochemistry, and Western blotting. It appears that endoplasmic reticulum (ER) stress is involved in this degeneration of Purkinje cells because ER stress-related substrates, such as CHOP and caspase 12, were strongly activated in Purkinje cells of pcd mice during the third postnatal (P) week. A significant increase in the expression of the ER-specific chaperone BiP suggested that unfolded protein responses were induced. We also found that Purkinje cells underwent apoptosis via the activation of caspase 3 and subsequent fragmentation of DNA. In addition to the activation of apoptosis in Purkinje cells, many activated microglial cells are found to be present in the molecular layer of the cerebellar cortex. In the later phase of degeneration, there was conspicuous expression of inducible nitric oxide synthase (iNOS), and some Purkinje cells were strongly labeled with an antibody to nitrotyrosine, suggesting that Purkinje cells in pcd mice are damaged by nitric oxide released from microglial cells. Administration of minocycline, which may inhibit iNOS expression, delayed the death of Purkinje cells in pcd mice and mildly improved their motor abilities. These findings suggest that ER stress participates in the degeneration of Purkinje cells and that activation of microglia accelerates Purkinje cell death in pcd mice. PMID- 16356647 TI - Regulation of the transcriptional activity of the tyrosine hydroxylase gene by androgen receptor. AB - Dopamine and the sex hormone testosterone are important factors regulating male sexual behavior. To investigate the possibility that these two factors are functionally interrelated, we investigated the potential role of the androgen receptor (AR) on transcriptional activity of the tyrosine hydroxylase (TH) gene that encodes the rate-limiting enzyme of the dopamine biosynthesis pathway. In this study, using transient co-transfection assays in TH-positive SK-N-BE(2)C and MN9D cells, we show that AR prominently transactivates TH promoter function in a ligand-dependent manner. Deletional and site-directed mutational analyses have mapped a putative androgen response element (ARE) in a region from -1562 to -1328 base pairs in the upstream TH promoter. We also found that DJ-1, one of recently identified genes whose mutations cause Parkinson's disease, down-regulated AR dependent TH activation by approximately 50% in SK-N-BE(2)C cells. Based on these data, we propose that AR activates TH gene expression and that DJ-1 may modulate AR activity as a transcriptional co-repressor. PMID- 16356648 TI - Regional brain cerebral glucose metabolism and temperament: a positron emission tomography study. AB - Personality, and in particular temperament, is thought to have a biological basis. In the present study, the relationships between regional brain glucose metabolism and temperament have been investigated. Regional brain glucose metabolism was measured using [18F] fluorodeoxyglucose positron emission tomography in 31 healthy subjects. Temperament was assessed using the Temperament and Character Inventory. Temperament dimensions were observed to be significantly correlated with specific brain regions. In particular, novelty seeking was significantly correlated with the superior temporal gyrus, inferior parietal lobule, and the precuneus, which have been reported to be related with impulsiveness, while reward dependence was significantly correlated with the caudate head, which has been shown to be associated with reward processing. The various aspects of temperament may have biological bases in the specific brain regions. The accumulation of results from studies of this kind should provide further evidence connecting personality traits with their biological bases. PMID- 16356649 TI - HLA-DRB1 genotyping in Italian migraine patients. AB - We examined the distribution of HLA-DRB1 alleles in a cohort 255 Italian migraine patients and in a control group of 325 healthy subjects. 214 patients fulfilled the ICHD-II criteria for migraine without aura and 41 patients the criteria for migraine with aura. The frequency of DRB1*16 allele was found to be significantly increased in migraine without aura patients (p=0.02; OR 1.97, 95% CI: 1.10-3.54) than in healthy controls. The frequencies of HLA-DRB1 alleles were not significantly different between migraine with aura patients and controls. We did not detect any effect of DRB1 alleles on age at onset, duration of the disease, frequency and duration of migraine attacks. Our data suggest the presence of a genetic susceptibility factor for migraine without aura within the HLA region. PMID- 16356650 TI - The effect of epigallocatechin gallate on suppressing disease progression of ALS model mice. AB - Epigallocatechin gallate (EGCG) is a constituent of green tea, and increasing evidence suggests that EGCG has neuroprotective effects on oxidative stress injured neuronal cells, especially motoneurons. Although the neuroprotective effects of EGCG have been demonstrated in Parkinson's and Alzheimer's diseases and ischemic stroke models, there has been no report on the effect of EGCG on an in vivo model of amyotrophic lateral sclerosis (ALS). This study was undertaken to evaluate the effect of EGCG on ALS model mice with the human G93A mutated Cu/Zn-superoxide dismutase (SOD1) gene. We treated each group of 11 ALS model mice with EGCG (1.5, 2.9, and 5.8 microg/g body weight), dissolved in 0.5 ml of 0.9% sterile NaCl, and one group of 11 with 0.5 ml of 0.9% sterile NaCl (control group) intraorally every day after 60 days of age (presymptomatic treatment). The treatment of more than 2.9 microg EGCG/g body weight significantly prolonged the symptom onset and life span, preserved more survival signals, and attenuated death signals. These data suggest that EGCG could be a potential therapeutic candidate for ALS as a disease-modifying agent. PMID- 16356651 TI - A pilot study of hormone modulation as a new treatment for mania in women with bipolar affective disorder. AB - We tested and compared the use of two adjunctive hormonal agents, tamoxifen and medroxyprogesterone acetate (MPA), for the treatment of acute mania or hypomania. A total of 13 women with acute Bipolar Affective Disorder in the manic or hypomanic phase were recruited from a clinical population to participate in this 28-day, three-arm, double blind, placebo-controlled study. The women who received tamoxifen exhibited significant improvement in symptoms of mania from baseline to final assessment compared with the placebo group. The MPA group improved more than the placebo group. Further exploration of tamoxifen as a useful adjunct in the treatment of acute manic symptoms in women with Bipolar Affective Disorder is warranted. PMID- 16356652 TI - Increased oxidative activity from hydrogen peroxide may be the cause of the predisposition to cataracts among patients with atopic dermatitis. PMID- 16356653 TI - Sprouting phenomenon, a new model for the role of A-beta fibers in wind up. AB - Wind up is a progressive frequency-dependent facilitation of the responses of nociceptive neurons observed on the application of repetitive (usually electrical) stimuli of constant intensity. The NMDA and NK1 receptors are essentially involved in wind up. After induction of wind up, stimulation of C fibers show the characteristics of wind up, but stimulation of Abeta fibers for induction of wind up is controversial. In this study, we have proposed a new model for the role of Abeta fibers in wind up, through sprouting of nerve fibers in the dorsal horn of spinal cord. We named it "sprouting phenomenon". It has been reported that in some clinical hyperalgesic states induced by peripheral injury or inflammation, wind up may aggravate the pain. For example, studies have indicated the presence of wind up in post-surgical states, some neuropathic pains, fibromyalgia syndrome, and post-herpetic neuralgia. According to sprouting phenomenon, it seems that some clinical interventions can be assessed to alleviate post-inflammatory pains: (1) Immediate and complete relief of inflammation by anti-inflammatory agents to prevent repetitive excitation of C fibers and subsequent morphological changes of dorsal horn laminae; (2) using local anesthetics in order to prevent pain signal transmission; (3) prevention of sprouting by intrathecal injection of some anti-proliferation agents; (4) using NMDA or NK1 receptor antagonists to prevent central mechanism of wind up. Some clinical trials have indicated the effectiveness of these antagonists. It is worth noting that future clinical studies are needed to validate these predictions. PMID- 16356654 TI - Migrainous scintillating scotoma and headache is ocular in origin: A new hypothesis. AB - Brain neuronal dysfunction has been implicated in pathogenesis of migraine but direct evidence is lacking. Scintillating scotoma of migraine is generally believed to originate at the visual cortex. While cortical spreading depression is a relatively late physiological alteration in migraine, its protective role in neuronal ischaemia is increasingly being recognized. Atenolol, nadolol, or verapamil prevent migraine but do not readily cross the blood-brain barrier or critically influence any brain or peripheral neuronal function. Typical migraine headache, aura, or scintillating scotoma has not been reported following enucleation or evisceration of the eye. In humans, pain and temperature fibres from only the ophthalmic division of the trigeminal nerve reach the upper cervical spinal segments. Pain in migraine attacks including occipital and nuchal discomfort reflects selective involvement of the ophthalmic nerve. Photophobia is largely a retinal reflex involving the ophthalmic division of the trigeminal nerve. Key clinical features of the migrainous scintillating scotoma are consistent with retinal origin. Spreading depression in the retina is well established. A subtle regional ocular sympathetic deficit prevails in migraine patients and possibly impairs regulation of intraocular choroidal blood volume and intraocular pressure. Several first-line migraine prophylactic agents lower the intraocular pressure. The neuro-ophthalmological basis for a monocular origin of migrainous scintillating scotomata due to mechanical deformation of the posterior segment of the corneo-scleral envelope consequent to choroidal venous congestion and rise in intraocular pressure is presented. Study of distribution and displaceability of the migrainous scintillating scotoma can settle its site of origin. Headache of migraine possibly arises from a similar mechanical deformation of the anterior eye segment followed by antidromic discharge in the trigeminovascular system. Lateralizing negative deficits such as homonymous hemianopia probably reflect vasospastic complications of migraine. A rational explanation for the most characteristic clinical features of migraine and a new template to elucidate the pharmacological basis of anti-migraine drugs is offered. PMID- 16356655 TI - Deficiencies in estrogen-mediated regulation of cerebrovascular homeostasis may contribute to an increased risk of cerebral aneurysm pathogenesis and rupture in menopausal and postmenopausal women. AB - Despite the catastrophic consequence of ruptured intracranial aneurysms, very little is understood regarding their pathogenesis, and there are no reliable predictive markers for identifying at-risk individuals. Few studies have addressed the molecular pathological basis and mechanisms of intracranial aneurysm formation, growth, and rupture. The pathogenesis and rupture of cerebral aneurysms have been associated with inflammatory processes, and these have been implicated in the digestion and breakdown of vascular wall matrix. Epidemiological data indicate that the risk of cerebral aneurysm pathogenesis and rupture in women rises during and after menopause as compared to premenopausal women, and has been attributed to hormonal factors. Moreover, experimental evidence supports a role for estrogen in the modulation of each phase of the inflammatory response implicated in cerebral aneurysm pathogenesis and rupture. While the risk of aneurysm rupture in men also increases with age, this increased risk has been attributed to other recognized risk factors including cigarette smoking, use of alcohol, and history of hypertension, all of which are more common in men than women. We hypothesize, therefore, that decreases in both circulating estrogen levels and cerebrovascular estrogen receptor density may contribute to an increased risk of cerebral aneurysm pathogenesis and rupture in women during and after menopause. To test our hypothesis, experiments are needed to identify genes regulated by estrogen and to evaluate gene expression and intracellular mechanisms in cells/tissues exposed to varying concentrations and duration of treatment with estrogen, metabolites of estrogen, and selective estrogen receptor modulators (SERMs). Furthermore, it is not likely that the regulation of cerebrovascular homeostasis is due to the actions of estrogen alone, but rather the interplay of estrogen and other hormones and their associated receptor expression. The potential interactions of these hormones in the maintenance of normal cerebrovascular tone need to be elucidated. Additional studies are needed to define the role that estrogen and other sex hormones may play in the cerebrovascular circulation and the pathogenesis and rupture of cerebral aneurysms. Efforts directed at understanding the basic pathophysiological mechanisms of aneurysm pathogenesis and rupture promise to yield dividends that may have important therapeutic and clinical implications. The development of non-invasive tools such as molecular MRI for the detection of specific cells, molecular markers, and tissues may facilitate early diagnosis of initial pathophysiological changes that are undetectable by clinical examination or other diagnostic tools, and can also be used to evaluate the state of activity of cerebral aneurysm pathogenesis before, during, and after treatment. PMID- 16356656 TI - Reciprocal oocyte sharing in return for altruistic sperm donation to fulfill the demands of clinical assisted reproduction and somatic cell nuclear transfer (SCNT) research. PMID- 16356657 TI - The C282Y mutation may have been positively selected as it mitigates the infertility of celiac disease. AB - Celiac disease and C282Y homozygous hemochromatosis have a similar increasing incidence across Europe. Both have gradients of frequency which increase from Turkey to North West Europe and culminate with a high frequency in Ireland. These two gradients follow the path of the Neolithic settlers who reached the edge of Europe at Ireland. Celiac disease and C282Y hereditary hemochromatosis have opposite effects on iron absorption and probably on the absorption of some other divalent metals including copper. The C282Y mutation is estimated to be some 2000 years old. Celiac disease is likely a much older disorder. The C282Y mutation may have been positively selected for as it increases absorption of divalent metal ions in celiac disease and thus has a mitigating effect on the infertility which may be associated with celiac disease. PMID- 16356658 TI - Comparative genomic hybridization pattern of non-anaplastic and anaplastic oligodendrogliomas--a meta-analysis. AB - Many oligodendrogliomas (ODG) have been investigated by comparative genomic hybridization (CGH). To visualize characteristic aberration profiles of non anaplastic in a comparison with anaplastic ODGs, we performed a meta-analysis of the CGH results of all 89 cases published so far. Therefore, we expanded all given aberrations to the maximum of 850 GTG band resolution. The frequencies of each chromosomal band affected by a genetic imbalance were calculated for WHO grades II and III separately. In non-anaplastic ODGs, -1p and -19q were the most prominent aberrations. In anaplastic ODGs, +7, -4q, -9p, -10, and -15q emerged additionally. We could confirm the existence of three disjunct genetically defined subgroups of ODGs, characterized by -1p/-19q (n=58, 65%, subgroup A), +7/ 10 (n=6, 7%, subgroup B) or the absence of either of the two patterns (n=25, 28%, subgroup C). Interestingly, we found a unique aberration pattern in subgroup C ( 1p31, -4q, -11p15, -18q, -22q, +17p, +17q) that was different from subgroups A and B, which could indicate a unique molecular carcinogenetic pathway of this ODG subset. Scrutinizing published putative progression markers of ODG, we found that only +7, -10, and -15q significantly correlated with a higher grade of malignancy. Summing up, the expansion of the CGH results to the 850 GTG band resolution enabled a meta-analysis to visualize WHO grade-specific aberration profiles in ODG for the first time. PMID- 16356659 TI - Attendance, drug use patterns, and referrals made from North America's first supervised injection facility. AB - BACKGROUND: North America's first government sanctioned supervised injection facility (SIF) was opened in Vancouver in response to the serious health and social consequences of injection drug use and the perseverance of committed advocates and drug user groups who demanded change. This analysis was conducted to describe the attendance, demographic characteristics, drug use patterns, and referrals made during the first 18 months of operation. METHODS: As part of the evaluation strategy for the SIF, information is collected through a comprehensive on-site database designed to track attendance and the daily activities within the facility. All users of the SIF must sign a waiver form and are then entered into a database using a unique identifier of their choice. This identifier is used at each subsequent visit to provide a prospective record of attendance, drug use, and interventions. RESULTS: From 10 March 2004 to 30 April 2005 inclusive, there were 4764 unique individuals who registered at the SIF. The facility successfully attracted a range of community injection drug users including women (23%) and members of the Aboriginal community (18%). Although heroin was used in 46% of all injections, cocaine was injected 37% of the time. There were 273 witnessed overdoses with no fatalities. During just 12 months of observation, 2171 individual referrals were made with the majority (37%) being referred for addiction counseling. INTERPRETATION: Vancouver's SIF has successfully been integrated into the community, has attracted a wide cross section of community injection drug users, has intervened in overdoses, and initiated over 2000 referrals to counseling and other support services. These findings should be useful for other settings considering SIF trials. PMID- 16356660 TI - Genomic structure and transcriptional studies on the mouse ribosomal protein S3 gene: expression of U15 small nucleolar RNA. AB - Ribosomal protein S3 (rpS3) is a multifunctional ribosomal protein (RP) which is known to function as a DNA repair endonuclease as well as an RP. Recently, it was reported that rpS3 is involved in apoptosis. We identified the complete 4760 base pair genomic structure of the mouse rpS3 gene, which is composed of 7 exons and 6 introns. Promoter study revealed that transcription of the mouse rpS3 gene started at two C residues embedded in the 5'-terminal oligopyrimidine tract (5' TOP); this was then compared with the human counterpart. Functional U15 small nucleolar RNAs (snoRNAs) were expressed from the first and the fifth introns. About 300 base pairs (bps) upstream of the 5'-untranslated region (5'-UTR) of the mouse rpS3 gene was sufficient to show maximum transcription activity. This report shows the conservation of the genomic structure of the rpS3 gene in vertebrates and characteristics of its promoter similar to those of promoters of other mammalian RPs. PMID- 16356661 TI - Isolation and characterization of retrotransposition-competent LINEs from zebrafish. AB - Long interspersed elements (LINEs) are a type of retroposon and are widely distributed in most eukaryotic genomes. LINEs are classified into two groups, the stringent type and relaxed type, based on the recognition of the 3' tail of their own RNA by reverse transcriptase (RT) during retrotransposition. Although most LINEs are thought to belong to the stringent type, retrotransposition studies of the stringent type LINEs are relatively limited compared with those of the relaxed type. We have now isolated two retrotransposition-competent LINEs (ZfL2-1 and ZfL2-2) from the zebrafish genome. Both ZfL2-1 and ZfL2-2 are members of the L2 clade; ZfL2-1 encodes two open reading frames (ORFs) and ZfL2-2 encodes one ORF, and each of the ORFs is required for retrotransposition. Using a retrotransposition assay in HeLa cells, we established that both ZfL2-1 and Zfl2 2 belong to the stringent type. We also demonstrated that an esterase (ES) domain encoded by ZfL2-1 ORF1 strongly enhances its own retrotransposition. The ES domain is encoded only in ORF1 of LINEs classified in the CR1 and L2 clades, although its function or significance in retrotransposition has not been elucidated. Thus, this is the first experimental evidence that the ES domain has an enhancing function during retrotransposition. These zebrafish LINEs will be useful for determining the function of ORF1 and the retrotransposition mechanism of stringent-type LINEs. PMID- 16356662 TI - Human polymorphism and human-chimpanzee divergence in pseudoautosomal region correlate with local recombination rate. AB - Previous studies have shown widespread correlation between nucleotide polymorphism and recombination rate, but the cause of this correlation is unresolved. One explanation is that recombination is associated with point mutations, potentially through mutagenic effects of meiotic crossover. This hypothesis predicts that regions of frequent recombination should show both elevated nucleotide diversity within a species and increased nucleotide divergence between species. Here we tested this hypothesis by studying the human short-arm pseudoautosomal region (PAR1), which recombines between X and Y chromosomes in men at a rate approximately 20 times the genome average. We sequenced dispersed intronic loci within PAR1 in a panel of humans and in the chimpanzee and directly measured sequence variation and recombination rate from these data. In line with previous reports, we saw a correlation between human polymorphism level and local recombination rate. Moreover, we also found a highly significant correlation between human-chimpanzee divergence and recombination rate. These results are consistent with the hypothesis that recombination is associated with point mutations, possibly because recombination is mutagenic. PMID- 16356663 TI - The evolution of introns in human duplicated genes. AB - In previous work [Jabbari, K., Rayko, E., Bernardi, G., 2003. The major shifts of human duplicated genes. Gene 317, 203-208], we investigated the fate of ancient duplicated genes after the compositional transitions that occurred between the genomes of cold- and warm-blooded vertebrates. We found that the majority of duplicated copies were transposed to the "ancestral genome core", the gene-dense genome compartment that underwent a GC enrichment at the compositional transitions. Here, we studied the consequences of the events just outlined on the introns of duplicated genes. We found that, while intron number was highly conserved, total intron size (the sum of intron sizes within any given gene) was smaller in the GC-rich copies compared to the GC-poor copies, especially in dispersed copies (i.e., copies located on different chromosomes or chromosome arms). GC-rich copies also showed higher densities of CpG islands and Alus, whereas GC-poor copies were characterized by higher densities of LINEs. The features of the copies that underwent the compositional transition and became GC richer are suggestive of, or related to, functional changes. PMID- 16356664 TI - Probing the secondary structure of salmon SmaI SINE RNA. AB - SmaI is a short interspersed element (SINE) of the salmon genome, and is derived from tRNA(Lys). We probed the secondary structure of SmaI SINE RNA by enzymatic cleavage and found that the RNA structure comprises three separate domains. The 5'-terminal region (the 5' domain) forms a tRNA-like cloverleaf structure, whereas the 3'-terminal region (the 3' domain) forms an extended stem-loop. The loop region is thought to be recognized by the reverse transcriptase (RT) encoded by the long interspersed element (LINE). The two structural domains are linked by a single-stranded region (the linker domain). Our melting profile analyses indicated the presence of two structural domains having different thermal stabilities, thus supporting the domain composition described above. Based on these results, we discuss the structural generality and evolutionary advantage of the domain composition of SINE RNA. PMID- 16356665 TI - Computational analysis of microRNA targets in Caenorhabditis elegans. AB - MicroRNAs (miRNAs) are endogenous approximately 22-nucleotide (nt) non-coding RNAs that post-transcriptionally regulate the expression of target genes via hybridization to target mRNA. Using known pairs of miRNA and target mRNA in Caenorhabditis elegans, we first performed computational analysis for specific hybridization patterns between these two RNAs. We counted the numbers of perfectly complementary dinucleotide sequences and calculated the free energy within complementary base pairs of each dinucleotide, observed by sliding a 2-nt window along all nucleotides of the miRNA-mRNA duplex. We confirmed not only strong base pairing within the 5' region of miRNAs (nts 1-8) in C. elegans, but also the required mismatch within the central region (nt 9 or nt 10), and we found weak binding within the 3' region (nts 13-14). We also predicted 687 possible miRNA target transcripts, many of which are thought to be involved in C. elegans development, by combining the above mentioned hybridization tendency with the following analyses: (1) prediction of the miRNA-mRNA duplex with free-energy minimization; (2) identification of the complementary pattern within the miRNA mRNA duplex; (3) conservation of target sites between C. elegans and C. briggsae, a related soil nematode; and (4) extraction of mRNA candidates with multiple target sites. Rigorous tests using shuffled miRNA controls supported these predictions. Our results suggest that miRNAs recognize their target mRNAs by their hybridization pattern and that many target mRNAs may be regulated through a combination of several specific miRNA target sites in C. elegans. PMID- 16356666 TI - Twelve novel C. elegans RNA candidates isolated by two-dimensional polyacrylamide gel electrophoresis. AB - C. elegans small RNAs (<50 nt) were separated by two-dimensional gel electrophoresis (2D-PAGE). cDNAs were prepared from the RNAs extracted from randomly chosen 2D-PAGE spots. Although many cDNA sequences corresponded to parts of known RNAs, twelve novel small RNA candidates were identified: eleven from 2D PAGE spots of the mixed-stage worm RNA preparation and one from those of the embryonic RNA preparation. These are encoded in the intergenic regions, in the introns of protein-coding genes, in the anti-sense strand of protein-coding sequences and repetitive sequence regions of the genome. None of them showed a characteristic structure of miRNAs, suggesting that they are candidates of other or new classes of RNAs. PMID- 16356667 TI - Tetrahydrogestrinone is a potent but unselective binding steroid and affects glucocorticoid signalling in the liver. AB - Tetrahydrogestrinone (THG) is a steroid recently identified to be misused as doping agent. However, the knowledge on functions of this substance in humans or animal models is rather limited. Therefore, it was our aim to further characterize the pharmacological profile of THG and identify potential adverse side effects. THG was synthesized, the purity was confirmed and its biological activity was tested. The potency of THG to transactivate AR dependent reporter gene expression was two orders of magnitude lower compared to dihydrotestosterone. THG binds with high affinity but unselective to the androgen (AR), progesterone (PR), glucocorticoid (GR) and mineralocorticoid (MR) receptor. Treatment of orchiectomised rats with THG resulted in a stimulation of prostate, seminal vesicle and levator ani muscle, indicating androgenic and anabolic properties. In the liver THG, in contrast to testosteronepropionate (TP), down regulates the expression of the GR dependent tyrosine aminotransferase gene (TAT). In summary, our results demonstrate that THG is not a specific AR agonist. THG exhibits a high binding affinity to all tested steroid hormone receptors and binds with highest affinity to the GR. Our in vivo data are indicative of an anabolic and androgenic potency of THG, but the repression of TAT demonstrates that THG also interferes with the glucocorticoid hormone system. Therefore, it is conceivable that an intake will result in adverse side effects. PMID- 16356669 TI - Increasing bioavailability of silymarin using a buccal liposomal delivery system: preparation and experimental design investigation. AB - Silymarin is a natural lipotropic agent of low bioavailability from oral products. The aim of our study is to prepare buccal liposomal delivery system of silymarin with higher bioavailability. The effect of lecithin:cholesterol molar ratio on the percentage drug encapsulated was investigated. The influence of fluctuating the amount of added drug was also determined. The effect of additives such as positive charge inducer, negative charge inducer and surfactants was studied using two different 2(3) full factorial designs. Furthermore, additives used to optimize liposomal product were also investigated for their optimal concentrations, release properties and in vitro permeation and absorption through chicken cheek pouch. Optimal liposomal encapsulation efficiency was found at 7:4 lecithin to cholesterol molar ratio. A decrease in entrapment efficiency with increasing cholesterol content was observed. Tween 20 or Tween 80 beyond 0.5 molar ratio decreased the entrapment efficiency. Positively charged liposomes showed superior entrapment efficiency over neutral and negatively charged liposomes. Release studies as well as permeation and absorption studies showed that hybrid liposomes prepared according to formula 3 containing lecithin, cholesterol, stearyl amine and Tween 20 in 9:1:1:0.5 molar ratio, respectively, gave the best drug absorption and permeation. It showed steady state permeation through chicken cheek pouch for 6h. This is expected to improve the bioavailability of silymarin in the developed liposomal buccal delivery system, as the results show an increase in drug penetration compared to free drug powder. PMID- 16356668 TI - Human hepatocytes are protected from ethanol-induced cytotoxicity by DADS via CYP2E1 inhibition. AB - We investigated the protective effects of diallyl disulfide (DADS), a potent inhibitor of cytochrome P450 2E1 (CYP2E1), on ethanol-induced toxicity in human hepatocytes. We found a clear dose-dependent response between ethanol and CYP2E1 activity. The ethanol-dependent CYP2E1 enzyme activity and protein expression, lactate dehydrogenase and aspartate transaminase release, malondialdehyde formation and caspase-3 activity decreased dramatically in the presence of DADS. Furthermore, DADS increased the hepatocellular glutathione (GSH) content and prevented the ethanol-dependent cellular GSH depletion. Our data show that DADS reduces ethanol-induced toxicity in human hepatocytes by reducing CYP2E1 activity and/or stabilizing the cellular GSH content, which might be of therapeutic interest. PMID- 16356670 TI - A comparative in vitro assessment of the drug release performance of pH responsive polymers for ileo-colonic delivery. AB - The aim of this study was to investigate the in vitro dissolution characteristics of pH-responsive polymers in a variety of simulated fluids. Prednisolone tablets were fabricated and coated with the following polymer systems: Eudragit S (organic solution), Eudragit S (aqueous dispersion), Eudragit FS (aqueous dispersion) and Eudragit P4135 (organic solution). Dissolution tests were conducted using a pH change method whereby tablets were transferred from acid to buffer. Three different buffer media were investigated: two compendial phosphate buffers (pH range 6.8-7.4) and a physiological buffer solution (Hanks buffer) with very similar ionic composition to intestinal fluid (pH 7.4). There was considerable drug release from tablets coated with Eudragit P4135 in acid, prompting discontinuation of further investigations of this polymer. Eudragit S (organic solution), Eudragit S (aqueous dispersion) and Eudragit FS on the other hand prevented drug release in acid, though subsequent drug release in the buffer media was found to be influenced by the duration of tablet exposure to acid. At pH 7.4 drug release rate from the polymer coated tablets was similar in the two compendial media, however in the physiological buffer, they were found to differ in the following order: Eudragit S (aqueous dispersion)>Eudragit FS>Eudragit S (organic solution). The results indicate that the tablets coated with the newer Eudragit FS polymer would be more appropriate for drug delivery to the ileo colonic region in comparison to the more established Eudragit S. More importantly, however, dissolution in the physiological buffer was found to be markedly slower for all the coated tablets than in the two compendial buffers, a result akin to reported slower dissolution of enteric coated tablets in vivo. There is therefore the need to adequately simulate the ionic composition of the intestinal fluid in the dissolution media. PMID- 16356671 TI - Skin targeted DNA vaccine delivery using electroporation in rabbits II. Safety. AB - The Achilles heel of gene-based therapy is gene delivery into the target cells efficiently with minimal toxic effects. Viral vectors for gene/DNA vaccine delivery are limited by the safety and immunological problems. Recently, nonviral gene delivery mediated by electroporation has been shown to be efficient in different tissues including skin. There are no detailed reports about the effects of electroporation on skin tissue, when used for gene/DNA vaccine delivery. In a previous study we demonstrated the efficacy of skin targeted DNA vaccine delivery using electroporation in rabbits [Medi, B.M., Hoselton, S., Marepalli, B.R., Singh, J., 2005. Skin targeted DNA vaccine delivery using electroporation in rabbits. I. Efficacy. Int. J. Pharm. 294, 53-63]. In the present study, we investigated the safety aspects of the electroporation technique in vivo in rabbits. Different electroporation parameters (100-300 V) were tested for their effects on skin viability, macroscopic barrier property, irritation and microscopic changes in the skin. Skin viability was not affected by the electroporation protocols tested. The electroporation pulses induced skin barrier perturbation and irritation as indicated by elevated transepidermal water loss (TEWL) and erythema/edema, respectively. Microscopic studies revealed inflammatory responses in the epidermis following electroporation using 200 and 300 V pulses. However, these changes due to electroporation were reversible within a week. The results suggest that the electroporation does not induce any irreversible changes in the skin and can be a useful technique for skin targeted DNA vaccine delivery. PMID- 16356673 TI - Cannabis seed identification by chloroplast and nuclear DNA. PMID- 16356672 TI - Anti-bacterial activity of some Brazilian medicinal plants. AB - Extracts from various organs of 25 plants of Brazilian traditional medicine were assayed with respect to their anti-bacterial activities against Escherichia coli, a susceptible strain of Staphylococcus aureus and two resistant strains of Staphylococcus aureus harbouring the efflux pumps NorA and MsrA. Amongst the 49 extracts studied, 14 presented anti-bacterial activity against Staphylococcus aureus, including the ethanolic extracts from the rhizome of Jatropha elliptica, from the stem barks of Schinus terebinthifolius and Erythrina mulungu, from the stems and leaves of Caesalpinia pyramidalis and Serjania lethalis, and from the stem bark and leaves of Lafoensia pacari. The classes of compounds present in the active extracts were determined as a preliminary step towards their bioactivity guided separation. No extracts were active against Escherichia coli. PMID- 16356674 TI - Application of hypervariable genetic markers to forensic identification of 'wild' from hatchery-raised red drum, Sciaenops ocellatus. AB - Forensic identification of 'wild' versus hatchery-produced (cultured) red drum (Sciaenops ocellatus), an economically important marine fish in the southern United States, was assessed using hypervariable nuclear-encoded microsatellites and sequences of mitochondrial DNA. Both genotype exclusion and likelihood-ratio tests successfully identified 'wild' and 'cultured' individuals within requisite error bounds and within the context of complete parental sampling. Of the two, genotype exclusion was more effective, producing satisfactory results with fewer microsatellites and larger allowable error rates. Assignment tests proved ineffective, most likely because of the low level of genetic divergence between the sampled populations. An optimal, minimum set of ten markers that will reduce potential genotyping costs is identified. Results of the study should allay concerns regarding identification of 'wild'-caught fish sold illegally. PMID- 16356675 TI - A newborn case of congenital laryngeal cyst complicated with pneumothorax and pneumomediastinum. AB - Benign congenital laryngeal cysts are rare entities. They often cause chronic hoarseness and severe stridor. Case reports of congenital laryngeal cyst complicated with pneumothorax and pneumomediastinum are very rare. A 3,112 g full term male newborn developed stridor which got worse during crying for 12 h after birth. Chest retractions were present with inspiration. Chest X-rays showed the presence of right pneumothorax and pneumomediastinum. Transnasal flexible laryngoscopic examination revealed a large cystic mass, which occupied almost the entire supraglottic airway. The operation was performed with the techniques of laryngomicrosurgery under general anesthesia. The cystic wall was punctured and serous liquid contents were aspirated. Excision of the entire cystic lesion was performed. The next day, extubation was performed without any troubles. The stridor had disappeared and the pneumothorax and pneumomediastinum were improved without further medical intervention. The histopathological examination revealed that the cystic wall consisted of normal squamous epithelial cells. It is reasonable to think that the high airway pressure due to congenital laryngeal cyst was responsible for pneumothorax and pneumomediastinum. PMID- 16356676 TI - LC/MS fingerprinting of Shenmai injection: a novel approach to quality control of herbal medicines. AB - Chromatographic fingerprinting has been recommended as a potential and reliable strategy for the quality control of herbal medicines. Although varieties of chromatographic techniques, particularly HPLC, have been widely employed, hyphenated chromatographic approach has not been sufficiently exploited in chromatographic fingerprinting. In this work, LC/MS fingerprinting of Shenmai injection was developed. Thirty ginsenosides as well as seven ophioponins were selected to construct the LC/MS fingerprint using selective ion monitoring (SIM) mode, while previous HPLC fingerprint [H.J. Zhang, Y.J. Wu, Y.Y. Cheng, J. Pharm. Biomed. Anal. 31 (2003) 175-183] only represents the ginsenosides. Subsequently, the proposed LC/MS fingerprints were applied to identifying the product manufacturers. All the samples were accurately classified based on their LC/MS fingerprints in conjunction with principal components analysis (PCA). This study would be potentially helpful to improve the quality control ability of fingerprinting-based strategy for complex herbal medicines. PMID- 16356677 TI - The moral nature of patient-centeredness: is it "just the right thing to do"? AB - OBJECTIVE: Patient-centeredness is regarded as an important feature of high quality patient care, but little effort has been devoted to grounding patient centeredness as an explicitly moral concept. We sought to describe the moral commitments that underlie patient-centered care. METHODS: We analyzed the key ideas that are commonly described in the literature on patient-centeredness in the context of three major schools of ethical thought. RESULTS: Consequentialist moral theories focus on the positive outcomes of providing patient-centered care. Deontological theories emphasize how patient-centered care reflects the ethical norms inherent in medicine, such as respect for persons and shared decision making. Virtue-based theories highlight the importance of developing patient centered attitudes and traits, which in turn influence physicians' behaviors toward their patients. CONCLUSION: Different ethical theories concentrate on different features of patient-centered care, but all can agree that patient centeredness is morally valuable. PRACTICE IMPLICATIONS: In order to sustain patient-centeredness as a moral concept, practitioners and students ought to examine these ideas to determine what their own personal reasons are for or against adopting a patient-centered approach. PMID- 16356678 TI - [The law passed on 4 March 2002, and its implications for medical practice]. PMID- 16356679 TI - [Infraclavicular block]. PMID- 16356680 TI - [Maintaining competence in pediatric anesthesia]. PMID- 16356682 TI - Molecular mechanisms of hydroxyurea(HU)-induced apoptosis in the mouse fetal brain. AB - Hydroxyurea (HU), a potent mammalian teratogen, affects proliferating embryonic cells and inhibits DNA synthesis. The teratogenic potential of HU has been well known in experimental animals for several decades. In this study, we investigated molecular mechanisms of HU-induced apoptosis in the telencephalon of the fetal brain by exposing pregnant mice to HU on day 13 of gestation. The number of TUNEL positive cells began to increase at 3 h, peaked at 12 h, and rapidly decreased at 24 h. Although changes of p53 mRNA expression were not observed by RT-PCR, a p53 positive reaction was detected immunohistochemically in the nuclei of neuroepithelial cells from 1 h to 6 h, and p53-protein expression was simultaneously identified by Western blot analysis. The expression of p53-target genes was detected at both the mRNA and protein. The mRNA levels of apotosis related genes (fas, fasL, and bax) and cell cycle-related genes (mdm2 and p21) were significantly elevated, and the degree to and sequence in which these target genes expressed was similar to those for fas, fasL, mdm2 and p21. Flow-cytometric and Western blot analyses of cell cycle-related proteins suggested that neuroepithelial cells are arrested at the S checkpoint from 3 to 6 h and at the G2/M checkpoint at 12 h, respectively. HU-induced apoptosis is considered to be mediated by p53 in the fetal brain. PMID- 16356683 TI - Observation of tumour thickness and resection margin at surgical excision of primary oral squamous cell carcinoma--assessment by ultrasound. AB - Tumour thickness and the status of resection margins are of prognostic significance in the treatment of oral cancer. In a single blind prospective study, 14 patients with biopsy proven oral squamous cell carcinoma had intraoral ultrasound imaging done preoperatively to measure tumour thickness, and intraoperatively to measure the deep surgical margin half way during resection. The cut surface was demonstrated on ultrasound by placing a metal, ultrasound reflective, retractor into the surgical cut. The ultrasound measurements were compared to the subsequent histological measurements. Using the threshold of 5mm as indicator of margin clearance, there was agreement in 10 out of 14 cases between ultrasound and histology. Ultrasound detection of close surgical margins had a sensitivity of 83% and a specificity of 63%. For preoperative tumour thickness measurement, ultrasound imaging showed a high degree of correlation with histology (Pearson correlation coefficient=0.95, P<0.01). This original paper demonstrates that high resolution ultrasound imaging applied intraorally is a reliable tool in objectively assessing both the tumour thickness and the surgical margin clearance at the time of surgery. PMID- 16356684 TI - Seizure anticipation, states of consciousness and marginal predictability in temporal lobe epilepsy. AB - PURPOSE: It has recently been shown that differences between the marginal predictability associated with scalp electrodes adjacent to and remote from the site of a seizure focus are able to distinguish between epochs temporally distant from and just prior to (within about 20 min) the onset of a seizure in patients with temporal lobe epilepsy. The purpose of this paper is to show that these differences of marginal predictability intervals are independent of the state of consciousness of the patient. METHODS: We have studied a data set encompassing 33 preictal epochs (within 1 h prior to a seizure) and 61 interictal epochs (defined as at least 1 h away from any seizure) from 14 patients. Each 30 s interval of each epoch was categorized into one of seven different states of consciousness. Statistical models were used to search for relationships (in aggregated data) between the values of differences of marginal predictabilities and state of consciousness. RESULTS: It was not possible to reject the null hypothesis of no relationship between differences of marginal predictabilities and state of consciousness. CONCLUSIONS: The values of the differences between marginal predictabilities on aggregated data are apparently insensitive to the state of consciousness. This conclusion, coupled with the fact that the differences between marginal predictabilities do depend on time to seizure, suggests the potential utility of these measures as the basis for ambulatory, non-invasive methods of seizure anticipation. However, the development of a practical non invasive method for seizure anticipation requires further extensive study on disaggregated data from individual patients. PMID- 16356685 TI - A ketogenic diet and knockout of the norepinephrine transporter both reduce seizure severity in mice. AB - Ketogenic diets (KD) have been known to be effective against epilepsy for more than 80 years, yet the mechanism(s) responsible for this action remain unknown. Norepinephrine (NE) has been shown to have anti-ictal effects against a wide variety of pro-convulsants and in animal models of epilepsy. Loss of noradrenergic activity is also associated with loss of the seizure protection seen following consumption of ketogenic diets. By contrast, knockout of the NE transporter (NET) gene, which elevates synaptic levels of norepinephrine, decreases seizure severity in mice fed normal diets. The purpose of this study was to compare the severity of maximal electroshock seizures in mice lacking the NET (NET KO) with that of wild type (WT) mice fed either a normal or a KD. In general, NET KO mice and mice fed a KD had a similar reduction in seizure severity, and the anticonvulsant effects of the genetic deletion of NET and the ketogenic diet were additive. These observations suggest that, while the noradrenergic system is required for the anti-seizure effects of the KD, additional mechanisms are involved. PMID- 16356687 TI - Hyaluronan forms complexes with low density lipoprotein while also inducing foam cell infiltration in the dermis. AB - BACKGROUND: Xanthoma is a foam cell infiltrating lesion similar to atherosclerosis. Glycosaminoglycans and proteoglycans have long been considered to play a role in atherogenesis. OBJECTIVE: The purpose of this study is to investigate the role of hyaluronan, the main dermal glycosaminoglycan, in xanthoma formation. METHODS: The complex formation of low density lipoprotein (LDL) with hyaluronan was investigated by assaying the cholesterol level of precipitates that were formed by incubating LDL, hyaluronan and cetylpyridinium chloride in the presence of Ca2+. The uptake of LDL by mouse peritoneal macrophages was studied by assaying the cellular cholesterol esterification activity. The responsible receptor for the LDL internalization was examined by saturating hyaluronan receptor and blocking class A macrophage scavenger receptor (CD204). Hyaluronan was injected into the dorsal skin of diet-induced hypercholesterolemic rabbits to reveal the xanthoma inducing activity of hyaluronan. RESULTS: Cetylpyridinium chloride precipitated hyaluronan, which had formed complexes with LDL. The macrophages incorporated hyaluronan-LDL complexes and oxidized LDL via CD204. Foam cell infiltration and cholesterol accumulation were induced by intradermal injections of hyaluronan in diet-induced hypercholesterolemic rabbits. CONCLUSION: Hyaluronan, like other sulfated glycosaminoglycans, retains LDL by forming a complex. Via macrophage scavenger receptors, macrophages incorporate not only LDL-hyaluronan complexes, but also oxidized LDL, which has been oxidized during the retention time. PMID- 16356688 TI - Ziprasidone-induced priapism requiring surgical treatment. AB - Both typical and atypical antipsychotic psychotropics have been reported to cause ischemic priapism presumptively secondary to alpha1-adrenergic blockade. This condition is a urologic emergency for if not evaluated and treated appropriately, long-term sequelae include erectile dysfunction and impotence. This paper addresses the first reported case of priapism requiring surgical intervention presumed to be secondary to ziprasidone. Increased clinician and patient education regarding priapism is required in order to minimize this adverse event and to maximize immediate treatment. PMID- 16356689 TI - Factor structure and cultural factors of disruptive behaviour disorders symptoms in Italian children. AB - OBJECTIVE: Poor parent and teacher awareness for attention-deficit/hyperactivity disorder (ADHD) and the scepticism of many clinicians on the prevalence of the disorder, make Italy an interesting environment in which to verify neuropsychological constructs generated in a predominantly Northern American cultural contest. The aim of the study was to verify, by factor confirmatory analysis, the empirical validity of the DSM constructs underlying the diagnostic criteria for developmental disruptive behaviour disorders in Italian school-age children. METHODS: Scores for DSM-IV inattention and hyperactivity/impulsivity, for oppositional defiant disorder (ODD) and for conduct disorder (CD) symptoms in 6-12 years old Italian children were analysed from 1575 parent and 1085 teacher forms of the disruptive behaviour disorders questionnaires collected in four different Italian regions. RESULTS: Reliability indicates high internal consistencies for both parent and teacher rating of inattention, hyperactivity/impulsivity, and oppositionality, but not for conduct problems. In accordance with the literature, a relatively low inter-rater convergent and discriminant validity correlation was observed comparing measures obtained by between parents and teachers. Confirmatory factor analysis of both parent and teacher data showed a better fit for a four-factor model, indicating a factor structure in accordance with the DSM-IV taxonomy. When completed by parents and teachers of clinically assessed ADHD, dyslexic or normal children, the disruptive behavioural disorder questionnaires showed a significant predictive diagnostic value. CONCLUSION: Although an informant variance higher that dimensional (trait) variance was observed, the study provides support for DSM-IV taxonomy for developmental disruptive disorders, showing construct validity of ADHD. ODD and CD could also be distinguished from each other. PMID- 16356690 TI - Exploring depression in schizophrenia. AB - BACKGROUND: A consistent amount of empirical research suggests that depression, besides interfering with quality of life and social functioning, may influence other symptom dimensions in schizophrenia, thus constituting an important domain for treatment strategies, outcome, and prognosis. AIM: This study investigated the factorial structure of the Calgary depression scale for schizophrenia (CDSS) in a sample of schizophrenic patients and explored the relationships between such factors, major symptom dimensions and subjective experiences. METHODS: One hundred and sixty-one subjects were examined to assess the severity of schizophrenic symptoms (scored according to the five-dimensional model of Toomey et al. [28]), the distress due to the subjective experience of negative symptoms, and the degree of subjectively-felt cognitive-affective vulnerability (i.e. basic symptoms). RESULTS: Principal component analysis revealed CDSS to include three main factors, namely: "depression-hopelessness" (factor I), "guilty idea of reference-pathological guilt" (factor II) and "early wakening" (factor III). Whereas the last factor did not correlate with any of the other psychopathological domains, the first two factors revealed multiple correlations with both diagnostic symptoms and subjective experiences. CONCLUSIONS: The results confirm the threefold factorial structure of the CDSS previously reported by the authors of the scale and could shed further light on the psychopathological nature of the components of depression in schizophrenia. The specific correlation patterns with diagnostic and subjective psychopatholgy substantiate the clinical distinction between a general depression factor ("depression-hopelessness") and a cognitive-guilt factor ("guilty idea of reference-pathological guilt"). PMID- 16356691 TI - Psychiatric side effects induced by supraphysiological doses of combinations of anabolic steroids correlate to the severity of abuse. AB - OBJECTIVE: The objective of our study was to evaluate the psychological consequences of real-world AAS use in athletes abusing such agents, in comparison with a placebo and control group of comparable athletes, while correlating the severity of abuse with the side effects observed. The hypothesis tested by the study was that the use of AAS induces a wide range of psychological side effects whose impact and emergence is dependent upon the severity of the abuse. DESIGN: The study includes a substantial group of AAS abusing athletes and two more groups demographically similar to the first, one composed of athletes not using any substance and a placebo group. All athletes were stratified according to the severity of AAS abuse. Psychometric instruments were applied to all athletes in specific time intervals, dependent to the AAS abusers' regimens, providing us with a final psychological profile that was to be compared to the pre-study profile. All results were comparable (within and between groups) for statistically significant differences and correlated to the severity of the abuse. Homogeneity of all groups was safeguarded by random doping controls, monitoring of drug levels and analysis of all self obtained drugs by method of liquid chromatography/mass spectrometry. All athletes were provided with a common exercise and dietary regime, so common training and nutritional conditions were achieved. METHODS: We studied a cohort of 320 body-building, amateur and recreational athletes, of whom 160 were active users of AAS (group C), 80 users administering placebo drugs (group B) and 80 not abusing any substance (Group A). Group C athletes were stratified according to AAS abuse parameters, thus providing us with three subgroups of "light, medium and heavy abuse". Athletes of groups A and B were included in a "no abuse" subgroup. The psychometric instruments used were the Symptoms Check List-90 (SCL-90) and the Hostility and Direction of Hostility Questionnaire (HDHQ). The psychometric evaluations took place within a time interval of 13 months. Statistical analysis was performed by using the Mann-Whitney/Wilcoxon two-sample non-parametric test (Kruskal-Wallis test for two groups) for data that were not normally distributed and Linear regression analysis was used to ascertain the correlation between severity of use and escalation of side effects. RESULTS: The study showed a statistically significant increase in all psychometric subscales recorded in group C, and no statistically significant difference in group C and A. There was a significant increase in the scorings of group C for all subscales of SCL-90 and HDHQ. Correlation of abuse severity and side effects showed that there was a statistical significant increase in Delta values of all SCL-90 and HDHQ subscales that escalated from light abuse to medium and heavy abuse/consumption patterns. CONCLUSIONS: The results of the study suggest that the wide range of psychiatric side effects induced by the use of AAS is correlated to the severity of abuse and the force of these side effects intensifies as the abuse escalates. PMID- 16356692 TI - The experience of carers of patients with severe mental illness: a comparison between London and Bologna. AB - PURPOSE: The experience of care giving inventory (ECI) is a self-reported measure of the caregiving experience as seen by the carer of a person with a serious mental illness. It comprises eight negative subscales and two positive subscales; positive and negative scores are obtained from these (ECI total positive and ECI total negative). This study aims to assess the validity and the reliability of the Italian version of the ECI and to compare the experience of carers in London and Bologna as measured by the ECI. METHOD: The sample consisted of 95 Bologna carers and 69 London carers of patients suffering from a functional psychotic disorder. The internal consistency of the Italian ECI and correlation between the Italian ECI total negative and the GHQ-12 (external validity) were estimated. Independent t-tests and chi-squared tests were used to compare sample characteristics and mean ECI scores. Regression analyses were used to control for confounders. RESULTS: Internal consistency and validity of the Italian ECI were similar to these previously found. The Bologna caregivers had a higher ECI total negative than London carers; this difference was partially explained by worse patients' functioning and by a higher proportion of parents in Bologna. Bologna carers scored significantly lower on ECI total positive after controlling for relationship and patient functioning. CONCLUSIONS: The Italian ECI is a well accepted and valid instrument to describe caregiving experiences. The similar scores in ECI total negative between London and Bologna underline a cross cultural consistency of negative experience of caregiving and re-emphasise the importance of patients functioning in different cultural contexts. The differences found for ECI total positive suggest area for further research. PMID- 16356693 TI - A survey of general practitioners' knowledge of symptoms and epidemiology of schizophrenia. AB - OBJECTIVE: To explore general practitioners' (GPs) knowledge of the symptoms and epidemiology of schizophrenia, and the GPs' characteristics associated with level of knowledge. METHOD: Survey questionnaires exploring practice in patients with early psychosis were mailed to all GPs in South-Western France (N=3829). GPs were asked to fill out questions anonymously on the symptoms and epidemiology of schizophrenia. RESULTS: The response rate was 23.6%. GPs had a fair theoretical knowledge of schizophrenia symptoms, but underestimated the prevalence and the risk of suicide. Recent attendance at a continuing medical education (CME) course on schizophrenia was the main predictor of level of knowledge on epidemiological figures, although its impact was relatively modest. Regarding risk factors, the implication of genetic factors was considered as null or modest by more than half of the GPs, while a large proportion of them stated that mother-baby interactional disturbances were frequently implicated in the aetiology of the disorder. CONCLUSION: GPs' level of knowledge on the public health impact and risk factors for schizophrenia needs to be improved in order to promote the delivery of accurate and de-stigmatising information to primary care patients. PMID- 16356694 TI - Cerebral metabolic consequences in the adult brain after neonatal excitotoxic lesions of the amygdala in rats. AB - In the present study the effects of neonatal excitotoxic lesions of the amygdala or ventral hippocampus on local cerebral glucose utilisation in the adult rat were studied by means of the [14C]2-deoxyglucose autoradiographic method. Our hypothesis was that damage to the brain during early development leads to long term functional activity changes in brain regions outside the primary lesioned area which might underlie the behavioural deficits observed in animals with neonatal brain damage. Cerebral glucose utilisation in animals with a neonatal amygdala lesion was significantly decreased in the amygdala itself and in several other brain regions. The neonatal ventral hippocampal lesion did not cause significant changes in cerebral glucose utilisation, except for a decrease in the primary damaged region (i.e. caudal ventral hippocampus). Behaviourally, animals lesioned in the amygdala displayed increased ambulatory activity both before and after puberty when exposed to a novel open field, while neonatal ventral hippocampal lesions did not affect adult exploratory behaviour as compared to sham controls. These results support our hypothesis that neonatal brain damage leads to long-term functional activity changes in brain regions outside the primary lesioned area. Moreover, they suggest that this long-term effect depends on the primary area lesioned since only damage to the amygdala, and not to the ventral hippocampus, affects the functional organisation of the brain of the animals later in life. Additionally, the findings may suggest that the functional changes in the brain may underlie the behavioural deficits observed after neonatal amygdala lesion in the rat. PMID- 16356695 TI - Diabetes mellitus and antipsychotic treatment in the United Kingdom. AB - OBJECTIVE: Treatment-emergent diabetes has been reported during exposure to conventional and atypical antipsychotics. This retrospective cohort study explored the UK General Practice Research Database (GPRD) to determine hazard ratios of diabetes for patients prescribed antipsychotics. METHODS: A Cox proportional hazard regression model using age, gender, and obesity (BMI > 30 kg/m2) was used to determine the hazard ratio (HR) of diabetes development in conventional antipsychotic (N = 59,089), atypical antipsychotic (N = 9053), individual antipsychotic, and general patient population cohorts (N = 1,491,548). RESULTS: Compared with the general GPRD patient population, patients exposed to conventional or atypical antipsychotics had a higher risk of developing diabetes (atypical antipsychotic cohort: HR = 2.9, CI = 2.0-4.4; and conventional antipsychotic cohort: HR = 1.9, CI = 1.6-2.3). The risk of developing diabetes during thioridazine, risperidone, or olanzapine treatment was significantly higher compared with the general GPRD patient population. CONCLUSION: Consistent with other epidemiology studies, this study supports an increased risk of developing diabetes during treatment with antipsychotics. PMID- 16356696 TI - Fluoxetine in the acute treatment and relapse prevention of combat-related post traumatic stress disorder: Analysis of the veteran group of a placebo-controlled, randomized clinical trial. AB - The efficacy and safety of fluoxetine (20-80 mg) was compared with placebo in 144 veterans [36.2 years], diagnosed with combat-related post-traumatic stress disorder (PTSD) selected from a 12-week acute and 24-week relapse prevention PTSD trial. In the acute phase, improvements were greater with fluoxetine than placebo in the disease-specific outcome measures: Treatment Outcome PTSD (TOP-8) total scores (SE):-9.05 (0.90) and -5.20 (1.23), p = 0.001; Clinician Administered PTSD Scale (CAPS) total scores:-31.12 (2.72) and -16.07 (4.24), p < 0.001; all CAPS subscores; Davidson Trauma Scale (DTS) total scores; and other general outcome measures. In the maintenance phase, fluoxetine was superior to placebo in sustaining improvement in TOP-8 [-1.01 (0.91) and 1.56 (0.95)] and CAPS [-4.93 (3.54) and 5.48 (3.66)]. The risk of relapse in the placebo arm was significantly greater than in the fluoxetine arm (log-rank test chi 2 = 4.090, df = 1, p = 0.048). Fluoxetine was well tolerated at a mean daily dose of 65 mg. PMID- 16356697 TI - High (20-Hz) and low (1-Hz) frequency transcranial magnetic stimulation as adjuvant treatment in medication-resistant depression. AB - Studies of repetitive transcranial magnetic stimulation (rTMS) in depression have found antidepressant effects when high frequency stimulation (HF-rTMS; >1 Hz) is applied over the left prefrontal cortex (LPF). A few studies have also reported success with low frequency stimulation (LF-rTMS) to the right prefrontal cortex (RPF). Both HF-rTMS and LF-rTMS have been reported to work better in areas with cerebral hypometabolism or hypermetabolism, respectively. Thirty medication resistant patients with major depression were randomized into three groups. The first group received sham rTMS and the second group received active rTMS (20-Hz rTMS to the LPF and 1-Hz rTMS to the RPF). The third group, however, received active rTMS that was focused on different regions of the brain after examination with single photon emission computed tomography (20-Hz rTMS to an area of relatively low activity and 1-Hz rTMS to an area showing relatively high activation). Patients and raters were blind to the treatment condition. Comparison of the sham rTMS group with the overall group that received active rTMS revealed statistically significant changes on the Hamilton Rating Scale for Depression after 10 sessions. This study demonstrated that combined 20+1-Hz rTMS was effective, but no additional advantages were obtained by focusing rTMS on areas identified by single photon emission tomography as showing high versus low levels of functional activity. PMID- 16356698 TI - Incorporation conditions guiding the aggregation of a glycosylphosphatidyl inositol (GPI)-anchored protein in Langmuir monolayers. AB - This work investigates the process of incorporation of a glycosylphosphatidyl inositol (GPI)-anchored alkaline phosphatase into Langmuir monolayers of dimyristoyl phosphatidic acid (DMPA). Three different methods of protein incorporation were assayed. When the protein solution was injected below the air water interface after formation of the lipid monolayer a micro-heterogeneous distribution of alkaline phosphatase throughout the interface was observed. Adsorption kinetics studied by fluorescence microscopy, associated with surface pressure measurements, led to the proposition of a model in which the protein penetration is modulated by the surface packing of the monolayer and intermolecular interactions occurring between the phospholipid and the protein. At initial surface pressures higher than 20 m Nm(-1), the protein is quickly adsorbed on the interface and the lateral diffusion drives the alkyl chains to turn towards the air phase while the polypeptide moiety faces the aqueous subphase. PMID- 16356699 TI - Micropatterning of polystyrene nanoparticles and its bioapplications. AB - Micropatterning of biomolecules forms the basis of cell culture, biosensor and microarray technology. Currently, the most widely used techniques are photoresist lithography, soft lithography or using robots which all involve multi-step surface modification directly on a planar substrate. Here we report a method to pattern biomolecules through self-assembling polystyrene nanoparticles in arrayed microwells on a solid surface to form well-ordered patterning, followed by attaching biomolecules to the assembled nanoparticles. The formation of colloidal patterns depends on capillary force, surface wettability and physical confinement. This method can be used for micropatterning a variety of biomolecules such as protein and antibody. PMID- 16356700 TI - Modulation of the rheological and mucoadhesive properties of thermosensitive poloxamer-based hydrogels intended for the rectal administration of quinine. AB - The aim of this work was to formulate and characterize thermosensitive gels based on poloxamer 407, a thermosensitive polymer, and hydroxypropylmethylcellulose (HPMC), a bioadhesive polymer, intended for the rectal delivery of quinine in children. In order to avoid the macroscopic phase separation between the two polymers it was necessary to add propanediol-1,2. After the formulation of stable ternary systems, their rheological properties were studied as a function of temperature. It was thus possible to obtained the gelation temperature of the systems as well as their elastic modulus, G', at 37 degrees C. It appeared that HPMC in the presence of propanediol-1,2 had a synergistic effect on the gelation of poloxamer 407. Tests on the rabbit rectal mucous membrane, founded on a technique of traction of the adhesive/adhered joint, made it possible to characterize the bioadhesive properties of the gels by measuring the work of adhesion, W, and the maximum detachment force, F(max). Over small concentration ranges of poloxamer and HPMC, very important variations of the viscoelastic and mucoadhesives properties were observed. It was shown that the viscoelastic and the bioadhesive properties were tightly correlated. Indeed, an empirical equivalence relation was established between the shear frequency and the rate of deformation in traction and allowed to describe all our samples by a master curve. This master curve would make it possible to predict the values of W at various rates of traction by the simple non-destructive measurement of the elastic modulus, G', at an equivalent shear frequency. PMID- 16356701 TI - A technical feasibility study of surfactant-free drug suspensions using octenyl succinate-modified starches. AB - Many new drugs exhibit poor wetting behaviour and low aqueous solubility. This is particularly an issue for preclinical studies like toxicological trials, in which considerably higher doses and volumes are being administered compared to clinical studies. Preclinical vehicles typically contain high levels of surfactants that can exert biological effects. However, the biological inertness of vehicles is pivotal for the application in preclinical studies stressing the need in finding new excipients to solve formulation problems of today's drug discovery. The present study investigated the technical feasibility of surfactant-free suspensions using a new poorly soluble drug as model. It was shown that octenyl succinate-modified starches adequately wetted the drug and homogenous tasteless suspensions were obtained. The polymer xanthan gum was identified as macroscopically compatible gelling agent. Concentration effects of xanthan, drug and different modified starches were studied in a D-optimal design with respect to rheological properties. The suspensions were also tested in an analytical centrifuge using NIR transmission profiles to obtain a measure of sedimentation stability under accelerated conditions. The modified starches exhibited only little influence on the viscosity as well as on the yield point in contrast to the rheological effects of xanthan gum. This gelling agent was the main stabilising excipient as the modified starches hindered to a lesser extent sedimentation. The most stable suspensions displayed convenient flow properties. The viscosity at 100 s(-1) and 25 degrees C was in technically acceptable range of 120-140 mPa s in view of a application via gavage or a syringe in animal studies. The results demonstrated that surfactant-free drug suspensions with excellent technical performance can be obtained using octenyl succinate-modified starches. The vehicles were tasteless and based on the experience of modified starches in the food industry, the vehicles should exhibit good tolerability. The future use of such surfactant-free drug suspensions in toxicological, pharmacokinetic and pharmacodynamic studies will have to determine their advantage in terms of biological inertness. PMID- 16356702 TI - Smoking cessation--but not smoking reduction--improves the annual decline in FEV1 in occupationally exposed workers. AB - INTRODUCTION: Individuals exposed both to cigarette smoke and respiratory pollutants at work incur a greater risk of development of airway hyperresponsiveness (AHR) and accelerated decline in forced expiratory volume in 1 s (FEV1) than that incurred by subjects undergoing each exposure separately. We examined whether smoking cessation or smoking reduction improves AHR and thereby slows down the decline in FEV1 in occupationally exposed workers. METHODS: We examined 165 workers (137 males and 28 females) participating in a smoking cessation programme. Nicotine tablets were used for smoking cessation or smoking reduction. Respiratory symptoms were assessed by questionnaire, FEV1 by spirometry and AHR by methacholine challenge test. At 1 year, subjects were classified into quitters, reducers, or continuing smokers. RESULTS: Sixty-seven subjects completed the study (32 quitters; 17 reducers; 18 continuing smokers). Respiratory symptoms improved markedly in quitters (P<0.001 for all comparisons) and less so in reducers (P values between 0.163 and 0.027). At 1 year, FEV1 had slightly but significantly improved in quitters (P=0.006 vs. smokers; P=0.038 vs. reducers) and markedly deteriorated in reducers and continuing smokers. Concurrent, 1-year change in AHR did not differ significantly among the groups. CONCLUSION: In occupationally exposed workers, stopping smoking markedly improved respiratory symptoms and, in males, slowed the annual decline in FEV1. Smoking reduction resulted in smaller improvements in symptoms but deterioration in FEV1. These findings were independent of AHR. While smoking cessation should remain the ultimate goal in workplace cessation programmes more studies are necessary to better ascertain the benefits of smoking reduction. PMID- 16356703 TI - Diversity of septin scaffolds. AB - Septins are ubiquitous GTP-binding proteins generally regarded as cytoskeletal components. Higher-order septin assemblies represented by the yeast septin collar function as cytoskeleton, providing structural support and scaffolds for many cellular factors. In metazoans, however, typical higher-order septin assemblies are often less predominant than dispersed 'low-order' septin populations. Recent studies revealed that septin populations with no obvious structure that had previously escaped our attention serve as scaffolds for kinetochore motor proteins and as sequestering depots for microtubule regulators. Unlike classic cytoskeletal polymers, which form uniform, continuous networks, septin polymers, being diverse, discontinuous and relatively static, seem suited to form discrete scaffolds. Thus, the septin system might be redefined as discrete scaffolds that are conditionally united to behave like cytoskeleton. PMID- 16356704 TI - Acute toxicity test of leachates from traditional and sustainable landfills using luminescent bacteria. AB - Landfilling is a fundamental step in any waste management strategy, but it can constitute a hazard for the environment for a long time. The need to protect the environment from potential landfill emissions makes risk assessment a decision tool of extreme necessity. The heterogeneity of wastes and the complexity of physical, chemical and biological processes that occur in the body of a landfill need specific procedures in order to evaluate the groundwater risk for the environment. Given the complexity of the composition of landfill leachates, the exact contribution of each potential toxic substance cannot be known precisely. Some reference contaminants that constitute the hazard (toxicity) of leachate have to be found to perform the risk assessment. A preliminary ecotoxicological investigation with luminescent bacteria has been carried out on different leachates from traditional and sustainable landfills in order to rank the chemicals that better characterize the leachate (heavy metals, ammonia and dissolved organic content). The attention has been focused on ammonia because it is present in high concentration and can last for centuries and can seriously contaminate the groundwater. The results showed that the toxicity of the leachate might reliably depend on the ammonia concentration and that the leachate toxicity is considerably lower in sustainable landfills where the ammonia had been degraded. This has an important consequence because if the containment system fails (as usually occur within 30-50yr), the risk of groundwater contamination will be calculated easier only in terms of the probability that the ammonia concentration is higher than a reference concentration. PMID- 16356705 TI - Changes in organic matter composition during composting of two digested sewage sludges. AB - Changes in the chemical and chemical-structural composition of the organic matter of two different sewage sludges (aerobic and anaerobic) mixed with sawdust (1:1 and 1:3, v/v) during composting were determined by monitoring chemical and microbiological parameters as well as by pyrolysis-gas chromatography. Composting was carried out in periodically turned outdoor piles, which were sampled for analysis 1, 30, 60 and 90 days after the beginning of the composting process. Both volatile organic matter and the water soluble C fraction decreased during composting, indicating that the more labile C fractions are mineralized during the process. Microbial activity as measured by microbial respiration (CO(2) evolved from compost samples during incubation) also decreased with composting, reflecting the more stable character of the resulting compost. No major differences were observed between the four composts studied as regards their chemical-structural characteristics. The acetonitrile, acetic acid and phenol pyrolytic fragment tended to increase with composting. Although the final composts were more aromatic in nature than the starting materials, a low degree of humification was observed in all four composts studied, as determined by their high proportion of polysaccharides and alkyl compounds. For this reason, the relationship between pyrolytic fragments, such as benzene/toluene or benzene+toluene/pyrrol+phenols, which are used as indices of humification for soil organic matter, are not of use for such poorly evolved sludge composts; instead, ratios that involve carbohydrate derivatives and aromatic compounds, such as furfural+acetic/benzene+toluene or acetic/toluene, are more sensitive indices for reflecting the transformations of these materials during composting. Both the chemical and microbiological parameters and pyrolytic analysis provided valuable information concerning the nature of the compost's organic matter and its changes during the composting process. PMID- 16356706 TI - Estimating the demand for municipal waste compost via farmers' willingness-to-pay in Ghana. AB - This paper has its primary focus on the analysis of perceptions and willingness to-pay (WTP) for composted municipal solid and faecal waste among urban and peri urban farmers and other potential compost users in Ghana. Participatory rural appraisal and contingent valuation methods (CVM) were used for the demand analysis. Most respondents were clear and firm in their responses to the principal question about WTP for compost, as well as in giving their views and perceptions about issues involved in demand for compost. The probit analysis proved valuable in highlighting variables, which explain variations in the WTP. The WTP analysis allowed the quantification of the compost demand under different scenarios of subsidized and non-subsidized compost production, with due allowance for a local reference price to cover compost station operating costs. The analysis revealed that the effective demand for compost for agricultural purposes is marginal and limited by farmers' transport costs. Only through the additional consideration of the demand of the construction sector can about 25% of the organic waste produced in Ghana's capital, Accra, be transformed and utilized. Public subsidies appear necessary and could be generated through savings in transport and disposal. Without subsidies, the challenge for an increased agricultural use is how to produce a low-cost but nutrient-rich compost, which can compete with abundant and cheap poultry manure and still achieve the price to maintain a compost station. The experience in Ghana shows that this is hardly possible except through private-public partnerships. PMID- 16356707 TI - A rapid highly-sensitive endotoxin detection system. AB - This paper presents a rapid, highly-sensitive, and low-cost method of endotoxin quantification based on the use of stress-responsive magnetoelastic sensors, that monitor the gel formation (viscosity change) of the Limulus Amoebocyte Lysate (LAL) assay in response to endotoxin. Ribbon-like magnetoelastic sensors, 12.7 mm x 6 mm x 28 microm, were immersed in a LAL assay after mixing with test samples of variable endotoxin concentration, and the decrease in resonance amplitude of the sensor was recorded as a function of time. Experimental results show excellent correlation between endotoxin concentration and the maximum clot rate, determined by taking the minimum point of the first derivative of the amplitude time curve, as well as the clotting-time, defined as the time that corresponds to the maximum clot rate. Using a LAL gel-clot assay with a sensitivity of 0.06 EU/ml (EU: endotoxin unit), the magnetoelastic sensor based technology can detect the presence of endotoxin at 0.0105 EU/ml in test requiring approximately 20 min. Unlike optical methods used for determining endotoxin concentration, the color of the test solution does not impact the magnetoelastic sensor measurement. Due to the small size of the sensor reader electronics and low cost, the magnetoelastic sensor based endotoxin detection system is ideally suited for wide-spread use in endotoxin screening for sepsis prevention. PMID- 16356708 TI - Detection of Escherichia coli O157:H7 with langasite pure shear horizontal surface acoustic wave sensors. AB - The toxigenic Escherichia coli O157:H7 bacterium has been connected with hemorrhagic colitis and hemolytic uremic syndrome, which may be characterized by diarrhea, kidney failure and death. On average, O157:H7 causes 73,000 illnesses, 2100 hospitalizations and 60 deaths annually in the United States alone. There is the need for sensors capable of rapidly detecting dangerous microbes in food and water supplies to limit the exposure of human and animal populations. Previous work by the authors used shear horizontal surface acoustic wave (SH SAW) devices fabricated on langasite (LGS) Euler angles (0 degrees, 22 degrees, 90 degrees) to successfully detect macromolecular protein assemblies. The devices also demonstrated favorable temperature stability, biocompatibility and low attenuation in liquid environments, suggesting their applicability to bacterial detection. In this paper, a biosensor test setup utilizing a small volume fluid injection system, stable temperature control and high frequency phase measurement was applied to validate LGS SH SAW biosensors for bacterial detection. The LGS SH SAW delay lines were fabricated and derivatized with a rabbit polyclonal IgG antibody, which selectively binds to E. coli O157:H7, in this case a non toxigenic test strain. To quantify the effect of non-specific binding (negative control), an antibody directed against the trinitrophenyl hapten (TNP) was used as a binding layer. Test E. coli bacteria were cultured, fixed with formaldehyde, stained with cell-permeant nucleic acid stain, suspended in phosphate buffered saline and applied to the antibody-coated sensing surfaces. The biosensor transmission coefficient phase was monitored using a network analyzer. Phase responses of about 14 degrees were measured for the E. coli detection, as compared to 2 degrees due to non-specific anti-TNP binding. A 30:1 preference for E. coli binding to the anti-O157:H7 layer when compared to the anti-TNP layer was observed with fluorescence microscopy, thus confirming the selectivity of the antibody surface to E. coli. PMID- 16356709 TI - Prognostic significance of combined expression of MUC1 and adhesion molecules in advanced gastric cancer. AB - The objective of the present study was to evaluate the combination of MUC1 and the status of adhesion molecules in advanced gastric cancers as a possible predictor of patient survival. Two hundred and two paraffin-embedded specimens of gastric carcinoma were examined by immunohistochemical staining using monoclonal antibodies against MUC1 mucin, E-cadherin and beta-catenin. The expression of MUC1 was considered positive if at least 10% of the neoplastic cells were stained. E-cadherin and beta-catenin were classified into four groups. Only a membranous pattern, which was stained as strongly as normal epithelial cells, was judged as normal. The absent pattern (loss of staining), cytoplasmic pattern (cytoplasmic staining with loss of membranous expression), and heterogeneous pattern (cytoplasmic staining with preservation of membranous expression) were considered abnormal. There was a significant relationship between MUC1-positive expression and abnormal expression of E-cadherin (P=0.017). The cancer with abnormal E-cadherin expression or MUC1-positive expression increased, indicating that the cancer invasion was deep. Survival analysis of the outcome revealed that the survival time for those with abnormal E-cadherin/MUC1-positive expression was shorter than for those with other expression patterns. Multivariate analysis revealed that patients with abnormal E-cadherin/MUC1-positive expression had a poorer prognosis with significance (P<0.0001). In conclusion, abnormal E cadherin/MUC1-positive expression pattern in advanced gastric cancer is an independent unfavorable prognostic marker. PMID- 16356710 TI - Effects of salinity on beta-carotene production by Dunaliella tertiolecta DCCBC26 isolated from the Urmia salt lake, north of Iran. AB - This study examined the effect of different salt concentrations (0.05-3 M of NaCl) on the kinetics of growth, total carotenoids and beta-carotene (all-trans and 9-cis) accumulated in Dunaliella tertiolecta DCCBC26, a microalgae strain isolated from the Urmia hypersaline lake, northwest of Iran. Results indicated that the highest amount of carotenoids detected (11.73 mg/l) was in the salinity of 0.5 M NaCl during the stationary growth phase. The percentage of the all-trans and 9-cis-beta-carotene in the exponential phase were 92% and 32% in salinities of 3 M and 0.5 M, respectively. However, only 23% of the beta-carotene was detected in the stationary growth phase of the microalgae in 0.5 M salinity and was 9-cis isomer. PMID- 16356711 TI - The protons of gluconic acid are the major factor responsible for the dissolution of tricalcium phosphate by Burkholderia cepacia CC-Al74. AB - Burkholderia cepacia CC-Al74 with a high ability for solubilizing tricalcium phosphate (TCP) was used to study the P-solubilization mechanism. We collected filtrates able to solubilize TCP from the cultures of strain CC-Al74 and demonstrated that the P-solubilization increased from 0 microg ml(-1) to 200 microg ml(-1) during exponential growth, when the pH decreased from 8 to 3. HPLC analysis revealed that the solubilization of TCP was mainly caused by the release of 16.3 mM gluconic acid. At this concentration, gluconic acid was capable of solubilizing 376 microg ml(-1) of TCP whereas water at pH 3 only solubilized 35 microg ml(-1). The difference is due to the final H+ concentrations which were 13.5 mM and 1 mM in 16.3 mM gluconic acid and deionized water, respectively at pH 3. PMID- 16356712 TI - Phytoextract-induced developmental deformities in malaria vector. AB - Larvicidal potential of petroleum ether (Pee), carbon tetrachloride (Cte) and methanol extract (Mee) of Artemisia annua, Chenopodium album and Sonchus oleraceus was observed against malaria vector, Anopheles stephensi Liston. The Pee of A. annua with LC50 16.85 ppm after 24 h and 11.45 ppm after 48 h of treatment was found most effective, followed by Cte of A. annua and Ch. album, Pee of Ch. album and Mee of A. annua. However, no significant larvicidal activity was observed in Mee of Ch. album and all the three extracts of S. oleraceous. The Pee of A. annua was further investigated for its effect on the metamorphosis and the development of the malaria vector. It influenced the early life cycle of An. stephensi by reducing the percentage of hatching, larval, pupal and adult emergence and also lengthening the larval and pupal periods. The growth index was also reduced significantly. As the extract has remarkable effect on the metamorphosis and high larvicidal potential, it could, therefore, be used as an effective biocontrol agent against the highly nuisant malaria vector. PMID- 16356713 TI - Protein tyrosine phosphatase 1B inhibitors from Morus root bark. AB - An organic layer prepared from the Chinese crude drug 'Sang-Bai-Pi' (Morus root bark) was studied in order to identify the inhibitory compounds for protein tyrosine phosphatase 1B (PTP1B). Bioassay-guided fractionation resulted in the isolation of sanggenon C (1), sanggenon G (2), mulberrofuran C (3) and kuwanon L (4) as PTP1B inhibitors, along with moracin O (5) and moracin P (6). Compounds 1 4 inhibited PTP1B with IC(50) values ranging from 1.6+/-0.3 microM to 16.9+/-1.1 microM. PMID- 16356714 TI - Synthesis of 3-benzyl-2-substituted quinoxalines as novel monoamine oxidase A inhibitors. AB - A new series of 3-benzyl-2-substituted quinoxalines have been synthesized by means of microwave enhancement of nucleophilic substitution reaction involving the corresponding 2-chloroquinoxaline analogs and substituted amines or hydrazine. The synthesized compounds were evaluated for their monoamine oxidase A and B inhibitory activity by determination of their IC(50). All the newly synthesized compounds showed more selective inhibitory activity toward MAO-A than MAO-B. In addition, the acute toxicity of the synthesized compounds was determined. This work may be a fruitful matrix of the synthesis of a new series of novel MAO-A inhibitors with good safety margins. PMID- 16356715 TI - A topological function based on spectral moments for predicting affinity toward A3 adenosine receptors. AB - The spectral moment descriptors have been applied to the study of affinity for A(3) adenosine receptors of 32 adenosine analogues. A model, able to describe more than 95% of the variance in the experimental activity, was developed with the use of the above-mentioned approach. The fragment contributions to the activity carried out show that the sulfonamido moiety at the N(6) position and hydrogen bonding play an important role in the interaction with the receptor. PMID- 16356716 TI - Do different surrogate methods detect lateral genetic transfer events of different relative ages? AB - Non-tree-based ("surrogate") methods have been used to identify instances of lateral genetic transfer in microbial genomes but agreement among predictions of different methods can be poor. It has been proposed that this disagreement arises because different surrogate methods are biased towards the detection of certain types of transfer events. This conjecture is supported by a rigorous phylogenetic analysis of 3776 proteins in Escherichia coli K12 MG1655 to map the ages of transfer events relative to one another. PMID- 16356717 TI - Trafficking arms: oomycete effectors enter host plant cells. AB - Oomycetes cause devastating plant diseases of global importance, yet little is known about the molecular basis of their pathogenicity. Recently, the first oomycete effector genes with cultivar-specific avirulence (AVR) functions were identified. Evidence of diversifying selection in these genes and their cognate plant host resistance genes suggests a molecular "arms race" as plants and oomycetes attempt to achieve and evade detection, respectively. AVR proteins from Hyaloperonospora parasitica and Phytophthora infestans are detected in the plant host cytoplasm, consistent with the hypothesis that oomycetes, as is the case with bacteria and fungi, actively deliver effectors inside host cells. The RXLR amino acid motif, which is present in these AVR proteins and other secreted oomycete proteins, is similar to a host-cell-targeting signal in virulence proteins of malaria parasites (Plasmodium species), suggesting a conserved role in pathogenicity. PMID- 16356718 TI - Adaptive evolution by optimizing expression levels in different environments. AB - Organisms adapt to environmental changes through the fixation of mutations that enhance reproductive success. A recent study by Dekel and Alon demonstrated that Escherichia coli adapts to different growth conditions by fine-tuning protein levels, as predicted by a simple cost-benefit model. A study by Fong et al. showed that independent evolutionary trajectories lead to similar adaptive endpoints. Initial mutations on the path to adaptation altered the mRNA levels of numerous genes. Subsequent optimization through compensatory mutations restored the expression of most genes to baseline levels, except for a small set that retained differential levels of expression. These studies clarify how adaptation could occur by the alteration of gene expression. PMID- 16356719 TI - Francisella tularensis travels a novel, twisted road within macrophages. AB - Francisella tularensis is a highly infectious intracellular bacterium that causes fulminating disease and is a potential bioweapon. Although entry of the bacteria into macrophages is mediated by novel asymmetric, spacious pseudopod loops, the nascent phagosome becomes tight fitting within seconds of formation. Biogenesis of the Francisella-containing phagosome (FCP) is arrested for 2-4h at a unique stage within the endosomal-lysosomal degradation pathway, followed by gradual bacterial escape into the cytosol, where the microbe proliferates. By contrast, other intracellular pathogens either proliferate within an idiosyncratic phagosome or escape within minutes into the cytoplasm to avoid degradation. Thus, trafficking of the FCP defies the dogma of classification of intracellular pathogens into vacuolar or cytosolic. The Francisella pathogenicity island and its transcriptional regulator MglA are essential for arresting biogenesis of the FCP. Despite sophisticated microbial strategies to arrest phagosome biogenesis within quiescent macrophages, trafficking of F. tularensis and other intracellular pathogens within interferon-gamma-activated macrophages is similar, in that the bacterial phagosomes fuse to lysosomes. The potential use of F. tularensis as a bioweapon has generated interest in the study of its molecular pathogenesis to identify targets for therapy, vaccination and rapid diagnosis. PMID- 16356720 TI - Non-conventional protein secretion in yeast. AB - Many proteins are transported to the cell surface of Saccharomyces cerevisiae and Candida albicans to be either integrated into the cell-wall structure or exported to the external medium. Secretion of many of these proteins through the classical endoplasmic reticulum-Golgi pathway is driven by a canonical N-terminal signal peptide. However, several surface proteins lacking this motif can also access the cell surface and remain loosely bound to the wall. The previous identification of these secretion-signal-less proteins in the cytoplasm as proteins that function as glycolytic enzymes, chaperones, translation factors and others suggests that they could be "moonlighting" (multifunctional) proteins. The accumulated evidence indicates that mechanisms of secretion other than the endoplasmic reticulum-Golgi pathway drive these proteins outside the plasma membrane. The relevance of these secretion-signal-less proteins in virulence and cell-wall dynamics warrants further characterization of alternative secretion in yeasts. PMID- 16356721 TI - Phase II study of two-weekly temozolomide in patients with high-grade gliomas. AB - Palliative chemotherapy has an increasing role in the management of recurrent high-grade gliomas. Temozolomide is a well-tolerated agent that results in objective responses and stabilisation of disease. Theoretically, temozolomide may be more effective when given in a prolonged schedule rather than the standard 5 days-monthly schedule. This Phase II study examined the efficacy and toxicity of temozolomide when given in a two-weekly schedule. Twenty-five patients received 150 mg/m2 temozolomide daily for seven days alternating with seven days of no treatment. One cycle of temozolomide was a total of two weeks treatment in every 28 days. Of the 25 evaluable patients, there was one complete response (4%), four partial responses (16%) and 10 patients had disease stablisation (40%). The progression free survival at 6 months was 56%. Two-weekly temozolomide was well tolerated with only four episodes of Grade 3 thrombocytopenia. Overall, two weekly temozolomide is an active and well tolerated schedule, but does not appear to improve on the activity of temozolomide using the standard 5-day schedule. PMID- 16356722 TI - The thiolase superfamily: condensing enzymes with diverse reaction specificities. AB - The formation of a carbon-carbon bond is an essential step in the biosynthetic pathways by which fatty acids and polyketides are made. The thiolase superfamily enzymes catalyse this carbon-carbon-bond formation via a thioester-dependent Claisen-condensation-reaction mechanism. In this way, fatty-acid chains and polyketides are made by sequentially adding simple building blocks, such as acetate units, to the growing molecule. A common feature of these enzymes is a reactive cysteine residue that is transiently acylated in the catalytic cycle. The wide catalytic diversity of the thiolase superfamily enzymes is of great interest. In particular, the type-III polyketide synthases make complicated compounds of great biological importance using multiple, subsequent condensation reactions, which are all catalysed in the same active-site cavity. The crucial metabolic importance of the bacterial fatty-acid-synthesizing enzymes stimulates in-depth studies that aim to develop efficient anti-bacterial drugs. PMID- 16356723 TI - Chutes and Ladders: the search for protein kinases that act on AMPK. AB - AMP-activated protein kinase (AMPK), a key regulator of energy homeostasis in mammalian cells, is, in turn, regulated by long-sought upstream protein kinases (AMPKKs). Following the recent identification of the tumor-suppressor kinase LKB1 as an AMPKK, a broader role for AMPK in metabolic economy has been unveiled by a new body of work from three groups that implicates the Ca(2+)/calmodulin dependent protein kinase kinases as AMPKKs. We suggest that PKE (protein kinase "energy" or "economy") is now an apt name for this kinase, which regulates both cellular and whole-organism energy homeostasis. PMID- 16356724 TI - Phosphoryl transfer in Ras proteins, conclusive or elusive? AB - The chemical mechanism of GTP hydrolysis by GTP-binding proteins of the Ras superfamily continues to inspire both experimental and computational biologists. The debate centres on the nature of the transition state, with arguments for both dissociative and associative, and whether there is a common GTPase mechanism for these proteins. In a recent structural analysis of Rab11, the product P(i) was found in an unusual configuration. This finding indicates that substrate-assisted catalysis might operate as a mechanism to enable nucleophilic attack in the intrinsic GTPase reaction, and would thus favour a pentavalent phosphorane intermediate. Recent findings on the GAP-mediated reaction of different Ras proteins suggest that a common mechanism might not exist and that G proteins probably show a continuum of electronic configurations in the transition state. PMID- 16356725 TI - Atomic level architecture of group I introns revealed. AB - Twenty-two years after their discovery as ribozymes, the self-splicing group I introns are finally disclosing their architecture at the atomic level. The crystal structures of three group I introns solved at moderately high resolution (3.1-3.8A) reveal a remarkably conserved catalytic core bound to the metal ions required for activity. The structure of the core is stabilized by an intron specific set of long-range interactions that involves peripheral elements. Group I intron structures thus provide much awaited and extremely valuable snapshots of how these ribozymes coordinate substrate binding and catalysis. PMID- 16356726 TI - The molecular basis of coupling of translocation and N-glycosylation. AB - Protein translocation and N-glycosylation are essential coordinated cellular processes that are mediated by the translocon and the oligosaccharyl transferase (OT), respectively. The recent identification of several specific interactions between the OT subunits and the translocon provides a molecular basis for the coupling of these two processes. Data suggest that multiple OT isoforms with different affinities for the translocon and ribosome and with heterogeneous subunit composition might exist in the endoplasmic reticulum (ER) membrane, thereby providing a means of regulating protein N-glycosylation. PMID- 16356727 TI - TiBS--how it all began. PMID- 16356728 TI - Synthesis and anti-inflammatory activity of a series of N-substituted naproxen glycolamides: nitric oxide-donor naproxen prodrugs. AB - A series of glycolamide naproxen prodrugs containing a nitrate group as a nitric oxide (NO) donor moiety has been synthesized. These compounds were evaluated for their anti-inflammatory activity, naproxen release, and gastric tolerance. Compounds 4a, 4b, 5a, 5b, 7b, and 7c exhibited anti-inflammatory activity equivalent to that of the parent NSAID, naproxen-Na, in the rat carrageenan paw edema model. At equimolar doses relative to naproxen-Na, the NO-donor glycolamide derivatives 4a, 4b, 5a, 5b, 7b, and 7c were gastro-sparing in the rat. Naproxen formation from these NO-donor glycolamides varied among the structures examined, with the N-substituent on the amide group having a particular influence, and demonstrated their prodrug nature. Compound 7b was selected for exemplary demonstration that the glycolamide nitrates can be bioactivated to release NO. These data open the possibility that naproxen glycolamide nitrates may represent a safer alternative to naproxen as anti-inflammatory medicines. PMID- 16356729 TI - Alpha-peptide/beta-sulfonamidopeptide hybrids: analogs of the chemotactic agent for-Met-Leu-Phe-OMe. AB - In order to gain information on the activity shown by alpha-peptide/beta sulfonamidopeptide hybrid analogs of the potent chemotactic agent fMLF-OMe, a structure-activity study is reported on N-Boc- and N-formyl tripeptide models containing an aminoalkanesulfonic acid as central residue. Directed migration (chemotaxis), superoxide anion production, and lysozyme release have been measured. The biochemical functions and the conformational properties of the new compounds are discussed and related to previously studied models containing beta residues. PMID- 16356730 TI - Design, synthesis, and biological evaluation of 1,3-diarylprop-2-en-1-ones : a novel class of cyclooxygenase-2 inhibitors. AB - A group of regioisomeric (E)-1,3-diarylprop-2-en-1-one derivatives possessing a COX-2 SO2Me pharmacophore at the para position of the C-1 or C-3 phenyl ring, in conjunction with a C-3 or C-1 phenyl (4-H) or substituted-phenyl ring (4-F, 4-OMe and 4-Me), were designed for evaluation as selective cyclooxygenase-2 (COX-2) inhibitors. These target (E)-1,3-diarylprop-2-en-1-ones were synthesized via a Claisen-Schmidt condensation reaction. In vitro COX-1/COX-2 isozyme inhibition structure-activity studies identified (E)-1-(4-methanesulfonylphenyl)-3-(4 methylphenyl)prop-2-en-1-one (9f) as a potent COX-2 inhibitor (IC50=0.3 microM) with a high COX-2 selectivity index (SI=106) comparable to that of the reference drug rofecoxib (COX-2 IC50=0.5 microM; COX-2 SI>200). A molecular modeling study where 9f was docked in the binding site of COX-2 showed that the para-SO2Me substituent on the C-1 phenyl ring is oriented in the vicinity of the secondary COX-2 binding site near Val523. The structure-activity data acquired indicate that the propenone moiety constitutes a suitable scaffold to design novel acyclic 1,3-diarylprop-2-en-1-ones with selective COX-2 inhibitory activity. PMID- 16356731 TI - Continuous flow PCB radiolysis with real-time assessment by gas chromatography. AB - We report the e-beam radiolysis of polychlorinated biphenyls (PCBs) in a continuous flow system, and the development of remote, real-time detection using gas chromatography. Aroclor-containing transformer oil was recirculated through an irradiation cell irradiated with 13-MeV electrons, at a flow rate of approximately 0.1L/min. The analytical equipment was protected from the beam by a 1.1-m concrete barrier. Data are presented showing the decrease in PCB concentration versus absorbed dose for pure transformer oil solution, and oil spiked with alkaline isopropanol to increase the PCB decomposition rate. Efficiency of PCB removal is expressed in terms of the dose constant, with units of reciprocal dose. PMID- 16356732 TI - A new visualization technique for the study of the accumulation of photoassimilates in wheat grains using [(11)C]CO(2). AB - Non-invasive real-time visualization of the accumulation of photoassimilates in the grains of an ear of wheat using [(11)C]CO(2) and positron emitting tracer imaging system (PETIS) was studied. [(11)C]CO(2) was supplied to the center of a fully expanded leaf of a wheat plant for an initial 10 min, and the transportation of (11)C-labeled photoassimilates into the grains of the ear was monitored for 120 min using the PETIS. Each grain was clearly identified in the obtained animation. The (11)C-labeled photoassimilates arrived at the ear from the [(11)C]CO(2)-absorbing leaf within 53 min from the time of supplying [(11)C]CO(2). After that, grains appeared on the image one by one from the basal part and full images of the grains appeared within 20 min. The time course of the accumulation of photoassimilates into each grain showed a different profile. Furthermore, the PETIS data suggested that the photo-condition of the ear plays an important role in the transportation of photoassimilates in wheat. PETIS can be used to visualize the dynamics of the substances in a living plant in real time and can exhibit the time course analysis of substances, such as the transportation, distribution, and accumulation. PMID- 16356733 TI - Traumatic mechanical injury to the hippocampus in vitro causes regional caspase-3 and calpain activation that is influenced by NMDA receptor subunit composition. AB - Apoptotic or necrotic cell death in the hippocampus is a major factor underlying the cognitive impairments following traumatic brain injury. In this study, we examined if traumatic mechanical injury would produce regional activation of calpain and caspase-3 in the in vitro hippocampus and studied how the mechanically induced activation of NR2A and NR2B containing N-methyl-d-aspartate receptors (NMDARs) affects the activation of these proteases following mechanical injury. Following a 75% stretch, significant levels of activated caspase-3 and calpain-mediated spectrin breakdown products were evident only in cells within the dentate gyrus, and little co-localization of the markers was identified within individual cells. After 100% stretch, only calpain activation was observed, localized to the CA3 subregion 24 h after stretch. At moderate injury levels, both caspase-3 and calpain activation was attenuated by blocking NR2B containing NMDARs prior to stretch or by blocking all NMDARs prior to stretch injury. Treatment with an NR2A selective NMDAR antagonist had little effect on either activated caspase-3 or Ab38 immunoreactivity following moderate injury but resulted in the appearance of activated caspase-3 in the dentate gyrus following severe mechanical stretch. Together, these studies suggest that the injury induced activation of NR2A containing NMDARs functions as a pro-survival signal, while the activation of NR2B containing NMDARs is a competing, anti-survival, signal following mechanical injury to the hippocampus. PMID- 16356734 TI - Distinct effects of p75 in mediating actions of neurotrophins on basal forebrain oligodendrocytes. AB - Previous studies indicate that brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF) and neurotrophin-3 (NT-3) increase myelin basic protein, (MBP) in differentiating basal forebrain (BF) oligodendrocytes (OLGs) (Du, Y., Fischer, T.Z., Lee, L.N., Lercher, L.D., Dreyfus, C. F., 2003. Regionally specific effects of BDNF on oligodendrocytes. Dev. Neurosci. 25, 116-126). While receptors, trk and p75, are expressed by subsets of oligodendrocytes (Du, Y., Fischer, T.Z., Lee, L.N., Lercher, L.D., Dreyfus, C. F., 2003. Regionally specific effects of BDNF on oligodendrocytes. Dev. Neurosci. 25, 116-126), those responsible for affecting differentiation have not been defined. In contrast, studies of peripheral Schwann cells reported that myelination is enhanced by BDNF working through p75, and diminished by trkC mediated processes (Cosgaya, J.M., Chan, J.R., Shooter, E.M., 2002. The neurotrophin receptor p75NTR as a positive modulator of myelination. Science 298, 1245-1248). To define receptors affecting central oligodendrocyte MBP, p75 knockout animals, p75 blocking antibodies, and an inhibitor of neurotrophin binding to p75, PD90780, were utilized. While p75 was implicated in the actions of NGF and NT-3, it did not affect actions of BDNF. On the other hand, K252a, an inhibitor of trk receptors, abolished the effects of the neurotrophins, including BDNF. All neurotrophins activated their respective trk receptors. PMID- 16356735 TI - High-level expression of human stem cell factor fused with erythropoietin mimetic peptide in Escherichia coli. AB - Stem cell factor (SCF) and erythropoietin are essential for normal erythropoiesis and induce proliferation and differentiation synergistically for erythroid progenitor cells. Here, we report our work on construction of SCF/erythropoietin mimetic peptide (EMP) fusion protein gene, in which human SCF cDNA (1-165aa) and EMP sequence (20aa) were connected using a short (GGGGS) or long (GGGGSGGGGGS) linker sequence. The SCF/EMP gene was cloned into the pBV220 vector and expressed in the Escherichia coli DH5alpha strain. The expression level of the fusion protein was about 30% of total cell protein. The resulting inclusion bodies were solubilized with 8 M urea, followed by dilution refolding. The renatured protein was subsequently purified by Q-Sepharose FF column. The final product was >95% pure by SDS-PAGE and the yield of fusion protein was about 40 mg/L of culture. UT 7 cell proliferation and human cord blood cell colony-forming assays showed that the fusion proteins exhibited more potent activity than recombinant human SCF, suggesting a new strategy to enhance biological activities of growth factors. PMID- 16356736 TI - Molecular cloning and characterisation of copper/zinc superoxide dismutase (Cu,Zn SOD) from the giant freshwater prawn Macrobrachium rosenbergii. AB - A copper/zinc superoxide dismutase (Cu,Zn-SOD) cDNA was cloned from the hepatopancreas of giant freshwater prawn Macrobrachium rosenbergii using reverse transcription-polymerase chain reaction (RT-PCR) by degenerate primers. Both 3'- and 5'-regions were isolated by the rapid amplification of cDNA ends method. Analysis of nucleotide sequence revealed that the Cu,Zn-SOD cDNA clone consists of 845 bp with an open reading frame of 603 bp encoding a protein of 201 amino acids with a 22 amino acid signal peptide. The calculated molecular mass of the mature proteins (179 amino acids) is 21 kDa with an estimated pI of 4.75. Two putative N-glycosylation sites, NXT and NXS, were observed in the Cu,Zn-SOD. Four conserved amino acids responsible for binding copper (H86, H89, H106 and H163) and four conserved amino acids responsible for binding zinc (H106, H114, H123 and D126) were observed. Sequence comparison showed that the Cu,Zn-SOD deduced amino acid sequence of M. rosenbergii has similarity of 60% and 64% to that of freshwater crayfish Pacifastacus leniusculus ecCu,Zn-SOD and blue crab Callinectes sapidus ecCu,Zn-SOD, respectively. Quantitative real-time RT-PCR analysis showed that Cu,Zn-SOD transcripts in haemocytes of M. rosenbergii increased 3h and 6h after injection of Lactococcus garvieae, whereas Cu,Zn-SOD transcripts decreased in the hepatopancreas 3h after L. garvieae injection. PMID- 16356737 TI - fMRI evidence for individual differences in premotor modulation of extrastriatal visual-perceptual processing of redundant targets. AB - To perceive the vast array of stimuli in the world around us, the visual system employs parallel processing mechanisms that ensure efficiency in perceiving multiple objects in a scene. A way to test this efficiency is to measure reaction time (RT) to pairs of identical stimuli, presented singly or as doublets; typically, the resulting phenomenon is the redundant targets effect (RTE), which manifests as faster RTs to paired than singly presented stimuli. It is controversial, however, whether the neural locus of the parallel processing mechanisms invoked to produce the RTE is perceptual or motor and why some studies observe a substantial RTE and others do not. To resolve these two issues, we measured the RTE in young adults while undergoing functional MRI. Regarding the question of a perceptual or motor basis for the RTE, we observed that bilateral activation of extrastriate cortex was prominent in paired vs. the sum of the two single stimulus conditions, indicating that the RTE invoked perceptual mechanisms; by contrast, the motor cortex was not disproportionately activated in this comparison. Regarding the magnitude of the RTE, we compared activation patterns in individuals with small vs. large RTEs and observed that frontal and premotor areas were activated with small RTEs. These data indicate that the primary processing level of response facilitation, observed as the RTE, is perceptual, but the modulation of the RTE magnitude is premotor and associated with basic aspects of response selection and preparation. PMID- 16356738 TI - The specificity of cortical region KO to depth structure. AB - Functional MRI studies have identified a cortical region designated as KO between retinotopic areas V3A/B and motion area V5 in human cortex as particularly responsive to motion-defined or kinetic borders. To determine the response of the KO region to more general aspects of structure, we used stereoscopic depth borders and disparate planes with no borders, together with three stimulus types that evoked no depth percept: luminance borders, line contours and illusory phase borders. Responses to these stimuli in the KO region were compared with the responses in retinotopically defined areas that have been variously associated with disparity processing in neurophysiological and fMRI studies. The strongest responses in the KO region were to stimuli evoking perceived depth structure from either disparity or motion cues, but it showed negligible responses either to luminance-based contour stimuli or to edgeless disparity stimuli. We conclude that the region designated as KO is best regarded as a primary center for the generic representation of depth structure rather than any kind of contour specificity. PMID- 16356739 TI - Development of Drosophila motoneurons: specification and morphology. AB - In the Drosophila ventral nerve cord, segmentally repeated sets of approximately 80 motoneurons are generated during embryogenesis. Within each hemisegment, each motoneuron is characterised by its axonal projection and innervation of a particular target muscle as well as its dendritic tree in the central nervous system. Codes of transcriptional regulators appear to specify in a hierarchical fashion the cell type, motoneuron sub-types and eventually unique cellular identities. Recent studies show that patterns of connectivity in the periphery are mirrored by patterns of dendritic arborisation centrally thereby providing a neuronal correlate of connectivity to the anatomy of the motor system in the periphery. While the principal mechanisms that underlie the development of the peripheral neuromuscular system have been studied in some detail, much less is known about how the dendrites and their patterns of connections develop in the CNS. PMID- 16356740 TI - Transcriptional regulation of mesendoderm formation in Xenopus. AB - Mesoderm and endoderm formation in Xenopus involves the coordinated efforts of maternally and zygotically expressed transcription factors together with growth factor signalling, including members of the TGFbeta and wnt families. In this review we discuss our current state of knowledge of these pathways, and describe in more detail some of the transcription factor-DNA interactions that are involved in mesendoderm formation. PMID- 16356741 TI - Trichinellosis: crossing the Chinese wall. PMID- 16356742 TI - The need for a new fluoroquinolone. PMID- 16356743 TI - The treatment of asthma in children: inhaled corticosteroids. AB - The evidence that asthma is characterized by extensive inflammation of the airways has warranted the use of inhaled corticosteroid (ICS) in asthma maintenance therapy. Corticosteroid treatment, especially if high or frequent doses are required, is associated with a range of adverse effects including adrenal suppression and impairment in growth and bone metabolism. New corticosteroids are in development, including mometasone furoate, and some of these are predicted to have reduced adverse effects such as the soft steroid ciclesonide. Soft steroids are designed for delivery near to their site of action, to exert their effect and then to undergo controlled and predictable metabolism to inactive metabolites. This review points out the anti-inflammatory effects of corticosteroid in asthmatic airways and the clinical efficacy and safety of ICS in asthmatic children. The development of a soft steroid should help to achieve the aim of improving the therapeutic profile of ICS in asthma and thus alleviate the ongoing problem of poor patient compliance especially in childhood. PMID- 16356744 TI - Characterization of baroreflex gain in the domestic pigeon (Columba livia). AB - Birds have a remarkable capacity to regulate circulation yet little is known about the avian baroreflex. Although both linear regression and curve-fitting techniques are frequently used to assess baroreflex function in mammals, only the former technique has been used in birds. We characterized baroreflex gain in domestic pigeons (Columba livia) and compared gain values derived from applying linear regression to ramp changes in mean arterial pressure (MAP) to values derived from fitting a four-parameter sigmoidal function to steady-state alterations in MAP. We found that, unlike mammals, pigeons do not display circadian patterns in MAP, HR or gain derived from bolus injections of vasoactive drugs. The pressor, but not depressor response, was attenuated by administration of the NMDA-antagonist ketamine, suggesting that central processing of the baroreflex may be similar in birds and mammals despite anatomical differences in arterial baroreceptive zones. Because graded infusions of vasoactive drugs could not consistently produce a plateau in the HR response, fitting data to a sigmoidal curve was difficult. Thus, we propose that variations of the Oxford method and linear regression analysis are superior method to assess baroreflex gain in pigeons than curve fitting. PMID- 16356745 TI - Lipid dynamics and plasma level changes of 17beta-estradiol and testosterone during the spawning season of gilthead seabream (Sparus aurata) females of different ages. AB - The present study was undertaken to evaluate whether the age of brood stock females of Sparus aurata affects the mobilization of lipids from muscle and liver towards the gonads to aid in oocyte development. Taking into account the role some hormones play in lipid mobilization the production of 17beta-estradiol (E2) and testosterone (T) was also measured throughout the spawning season. Four groups of fish were established consisting of 3-, 4-, 5- and 6-year-old females (1.3, 1.5, 2.3 and 2.8 kg average weight, respectively), maintained in separate tanks together with a number of two-year-old males. The results for all four groups showed no variations in fish total length between the beginning and end of the experimental period. However, losses were recorded both of body weight and condition factor. In general terms, there were no significant differences in the plasma levels of E2, T and the gonadosomatic (GSI) and hepatosomatic (HSI) index among the four groups throughout the spawning season. In all groups, the plasma levels of E2, T, GSI and HSI were at their lowest at the end of spawning. Between December (pre-spawning) and March (mid-spawning), all groups suffered depletion of the lipid content of liver and muscle, while gonad lipid content remained constant. The mobilization of lipids from liver and muscle to contribute to the upkeep of the gonadal lipid involved the mobilization of all the fatty acid groups, saturates, monoenes, n-6, n-3, and n-3 HUFA. A good correlation existed between the percentage of the various fatty acid groups transferred from muscle and the age of the reproductive females. However, the greater lipid mobilization from muscle matching the greater age of the reproductive females did not result in greater lipid gonadal reserves or greater body size, suggesting that reproduction on the part of older females requires greater effort. Despite this, the results as a whole indicated that lipogenic capacity, considered as the mobilization of lipids from muscle and liver towards the gonad for the development of oocytes, is unaffected by age in reproductive 3- to 6-year-old gilthead seabream females. PMID- 16356746 TI - Influence of temperature and gonadal steroids on the ontogenetic expression of brain serotonin 1A and 1D receptors during the critical period of sexual differentiation in tilapia, Oreochromis mossambicus. AB - cDNA sequences of serotonin (5-hydroxytryptamine, 5-HT) 1A and 1D receptors were cloned from the tilapia, Oreochromis mossambicus, brain. The influence of both gonadal steroids and temperature on the ontogenetic expression of brain 5-HT1A and 5-HT1D receptors from days 5 to 15 post-hatch, a critical period of sexual differentiation, was investigated using quantitative real-time reverse transcription-polymerase chain reaction. Neither estrogen nor methyltestosterone had an effect on the ontogenetic expression of 5-HT1A or 5-HT1D receptors. Between days 5 and 10 post-hatch, a critical period for low-temperature-induced feminization, we found no significant difference in the ontogenetic expression of 5-HT1A between exposure to low and elevated temperature. A similar result was found for 5-HT1D. Between days 10 and 15 post-hatch, a critical period for elevated-temperature-induced masculinization, the ontogenetic expression of neither brain 5-HT1A nor 5-HT1D was altered by exposure to elevated temperature. These results suggest that neither brain 5-HT1A nor 5-HT1D plays a critical role in either gonadal steroid- or temperature-induced sexual differentiation. PMID- 16356747 TI - NO-cGMP signaling in development and stem cells. AB - Despite the recognition that the NO-cGMP signaling pathway is involved in so many physiological and pathological events, a clear understanding of many of the functions of this signaling pathway remains elusive. Because of its pleiotropic and often transient actions, its modulation for therapeutic purposes in multiple pathological states is a complex issue. Recent work that combines the areas of developmental and stem cell biology and NO-cGMP signaling in various models may help us elucidate some of these functions and even discover novel actions for this signaling paradigm. This review will discuss some of the recent work in these areas, with additional focus on the nitric oxide receptor, soluble guanylyl cyclase. PMID- 16356748 TI - DNA polymorphisms and haplotypes in the 5'-upstream region of the ABO blood group gene. AB - We investigated the polymorphisms in the 5'-upstream region between nucleotide position (nt) -9600 and nt-4105 of the ABO blood group gene using PCR and direct sequencing methods. We found 16 single nucleotide polymorphisms, two insertion deletion polymorphisms and two sequence polymorphisms. One of the insertion deletion polymorphisms was found at nts from -9605 to -9204, and the alleles of that locus were named ABORR*L (non-deletion) and ABORR*S (52-base-deletion). There were two haplotypes constructed from 14 polymorphisms in the region between nt-9600 and nt-7565; they were tentatively named ABORR*L-associated and ABORR*S associated haplotypes. The ABORR*L-associated haplotype may link with ABO*O(A) (also known as ABO*O201), and the ABORR*S-associated haplotype links with the other common alleles. This indicates the existence of two major lineages of the Japanese ABO alleles in the 5'-upstream region from nt-7565. In contrast to these findings, we observed six haplotypes in the region between nt-6371 and nt-4105, and we assume that the sequence in that region is variable as compared with those in the other 5'-upstream regions. We examined the generation of two sequence polymorphisms found in the present study. In the both cases, the formation of a hairpin loop caused by palindrome in a single-stranded DNA molecule may play an important role in generating the polymorphism. PMID- 16356749 TI - Cafe coronary syndrome-fatal choking on food: an autopsy approach. AB - To examine the characteristic features of fatal food asphyxia and to develop an autopsy approach to such cases a retrospective study of autopsy files was undertaken at Forensic Science SA (Adelaide, Australia) over a 10 year period from 1993 to 2002 for all cases of food asphyxia/cafe coronary syndrome. Forty four cases were identified (M;F=21:23), with one infant (11 mths) and 43 adults (30-96 yrs; mean 68.9 yrs), with a preponderance of victims (57%) aged between 71 and 90 yrs. Deaths occurred in nursing homes (N=22) cases, at home (N=11) and in restaurants (N=4). Twenty-seven of the victims (61%) had histories of neurological or psychiatric disorders such as dementia (N=8), schizophrenia (N=6), Alzheimer disease (N=4), atherosclerotic cerebrovascular disease (N=4), mental impairment (N=2), multiple sclerosis (N=1), Parkinson disease (N=1) and obsessive-compulsive disorder (N=1). Twenty-seven cases (61%) were described as either edentulous or having significant numbers of teeth missing. Toxicological evaluation of blood revealed alcohol and a variety of psychotropic prescription medications in 19 cases. Sudden collapse during or shortly after a meal should always raise the possibility of cafe coronary and the autopsy examination should not only attempt to demonstrate airway occlusion by a bolus of food, but also to identify or exclude underlying neurological disease. Such cases may raise issues concerning adequacy of care and appropriateness of medication. The diagnosis of cafe coronary syndrome can only be made with confidence after the clinical history and circumstances of death have been clearly established, impacted material has been demonstrated in the airway at autopsy (or recorded by those attempting resuscitation), risk factors have been identified and other possible causes of death have been excluded. PMID- 16356750 TI - Brown Sequard syndrome due to cervical pen assault. AB - Brown Sequard syndrome is an uncommon incomplete spinal cord injury with ipsilateral motor and proprioception loss and contralateral pain and temperature loss. A 14-year-old student with classic Brown Sequard syndrome after pen assault injury by his classmate in posterior neck area presented here; with medical and supportive care, the patient was discharged with good outcome. Brown Sequard syndrome is mostly due to penetrating trauma. Several types of penetrating objects have been reported but this appears to be the first report of a pen or pencil injury. PMID- 16356751 TI - A study of blood and urine alcohol concentrations in cases of alleged drug facilitated sexual assault in the United Kingdom over a 3-year period. AB - This paper details the alcohol concentrations found in a selection of 1,014 cases of claimed drug-facilitated sexual assault analysed at The Forensic Science Service, London Laboratory between January 2000 and December 2002. Where appropriate, either a whole blood sample and/or a urine sample was analysed for alcohol, common drugs of abuse and potentially stupefying drugs. The samples were collected from a complainant within 12 h of an alleged incident in 391 of the 1014 cases analysed. Of these, the majority (81%) contained alcohol. The presence of alcohol itself was not surprising as most of the alleged incidents were associated with social situations such as at a public house, bar, night-club or party, where it is expected that alcohol would have been consumed. However, 233 (60%) of the 391 cases had a high back-calculated figure, where high is defined as greater than 150 milligrams per 100 millilitres (150 mg%). Some of these samples were also found to contain illicit drugs. This is the first paper to our knowledge which discusses in detail the significance of the alcohol concentrations found in cases of this type. PMID- 16356752 TI - Profile of unnatural deaths in Manipal, Southern India 1994-2004. AB - Pattern of unnatural deaths is a reflection of the prevailing social set up and mental health status of the region. In an attempt to understand the magnitude and pattern of unnatural deaths in Manipal, Southern India, a 11-year retrospective study was carried out in the Department of Forensic Medicine, Kasturba Medical College, Manipal. The study revealed a rise in road traffic fatalities (37%) that constituted the majority of autopsies carried out followed by death due to poisoning and burns. Male preponderance was quite evident (2.5:1) except for death due to burns where ratio was reversed (1:2.9). People in 3rd decade were most prone to such fatalities. Of the total medicolegal autopsies conducted in this period, the manner of death was unnatural in 98% of the cases and 71% of them were accidental in nature. PMID- 16356753 TI - Invasive aspergillosis in association with criminal arsenic poisoning. AB - A 26-year-old man suffered acute arsenic poisoning after a poisoning attempt. He developed multiple organ failure including encephalopathy, bleeding disorders, pancreatitis, renal and hepatocellular impairment. Generalized erythroderma also developed within one week after admission. The developed acute respiratory distress syndrome and Aspergillus fumigatus was isolated from the endotracheal aspirate. Despite intensive care support, antidote administration and various epuration techniques, the patient died on day 26 from subarachnoid bleeding. An autopsy was obtained and the concentration of arsenic was determined in different tissues. Multiple abscesses due to Aspergillus fumigatus were seen in the lungs, myocardium and kidneys. This uncommon complication in a previously immunocompetent patient could be related to impaired immunity directly caused by arsenic poisoning. PMID- 16356754 TI - The oncogenic serine/threonine kinase Pim-1 directly phosphorylates and activates the G2/M specific phosphatase Cdc25C. AB - The proto-oncogene Pim-1 encodes a serine-threonine kinase which is a downstream effector of cytokine signaling and can enhance cell cycle progression by altering the activity of several cell cycle regulators among them the G1 specific inhibitor p21(Waf), the phosphatase Cdc25A and the kinase C-TAK1. Here, we demonstrate by using biochemical assays that Pim-1 can interact with the phosphatase Cdc25C and is able to directly phosphorylate the N-terminal region of the protein. Cdc25C is functionally related to Cdc25A but acts specifically at the G2/M cell cycle transition point and can be inactivated by C-TAK1-mediated phosphorylation. Immuno-fluorescence experiments showed that Pim-1 and Cdc25C co localize in the cytoplasm of both epithelial and myeloid cells. We find that phosphorylation by Pim-1 enhances the phosphatase activity of Cdc25C and in transfected cells that are arrested in G2/M by bleomycin, Pim-1 can enhance progression into G1. Therefore, we propose that Pim-1 activates Cdc25C by a direct phosphorylation and can thereby assume the function of a positive cell cycle regulator at the G2/M transition. PMID- 16356755 TI - Cell wall proteins: a new insight through proteomics. AB - Cell wall proteins are essential constituents of plant cell walls; they are involved in modifications of cell wall components, wall structure, signaling and interactions with plasma membrane proteins at the cell surface. The application of proteomic approaches to the cell wall compartment raises important questions: are there technical problems specific to cell wall proteomics? What kinds of proteins can be found in Arabidopsis walls? Are some of them unexpected? What sort of post-translational modifications have been characterized in cell wall proteins to date? The purpose of this review is to discuss the experimental results obtained to date using proteomics, as well as some of the new questions challenging future research. PMID- 16356756 TI - Genetically engineered cytoplasmic male sterility. AB - Cytoplasmic male sterility, conditioned by some maternally inherited plant mitochondrial genomes, is the most expedient method to produce uniform populations of pollen-sterile plants on a commercial scale. Plant mitochondrial genomes are not currently amenable to genetic transformation, but genetic manipulation of the plastid genome allows engineering of maternally inherited traits in some species. A recent study has shown that the Acinetobacter beta ketothiolase gene, expressed in the Nicotiana tabacum plastid, conditions maternally inherited male sterility, laying the groundwork for new approaches to control pollen fertility in crop plants. PMID- 16356757 TI - Ethanol-inducible gene expression: non-transformed plants also respond to ethanol. PMID- 16356758 TI - Apical-basal polarity: why plant cells don't stand on their heads. PMID- 16356759 TI - Phloem: the long and the short of it. AB - The structure of phloem sieve-element-companion-cell complexes reflects a duality of function: to conduct photoassimilates throughout the plant, and to exchange solutes between the phloem and surrounding tissues. The conceptual integration of these long- and short-distance functions requires the abandonment of a long cherished concept in phloem physicochemistry, that source-sink turgor differentials control flow. The manifest inability of decentralized organisms such as plants to control phloem translocation centrally disqualifies such differentials as control variables; besides, the phloem is maximally efficient if the pressure differentials are small. Testing this hypothesis and whether turgor differentials are small will require a significant recommitment to studying the quantitative anatomy of phloem. PMID- 16356760 TI - Tailoring Ras-pathway--inhibitor combinations for cancer therapy. AB - Constitutive activation of Ras pathways plays a critical role in cancer development and maintenance. Inhibitors of such pathways are already in use for cancer therapy, with significant but as yet only partial success in the most deadly types of human cancers, against which even combinations of Ras-pathway inhibitors with classic cytotoxic drugs or irradiation are insufficient. Combinations of farnesyl transferase inhibitors (FTI's), inhibitors of Ras pathways, are now in use in clinical trials. In this review we analyze possible reasons for the limited efficacy--including the diverse and sometimes even contradictory effects of active Ras pathways in tumor cells--and propose possible alternative methods of tailoring Ras-pathway inhibitor combinations for cancer therapy. Such tailoring is now possible thanks to increased knowledge of the complexity of Ras pathways, their cooperation with other oncogenic pathways, and their "addictive" nature. We provide examples demonstrating that this knowledge can be translated into useful drug combinations that disrupt multiple oncogenic pathways and hit a weak point of a given tumor cell. One such example is combination treatment with a Ras inhibitor and a glycolysis blocker for pancreatic tumor cells. The future design of such potential drug combination therapies and the follow-up of their outcome will undoubtedly be facilitated by gene-expression profiling and proteomic methods. PMID- 16356762 TI - Towards the control of Echinococcus multilocularis in the definitive host in Japan. AB - Echinococcus multilocularis is distributed all over Hokkaido, the northern island of Japan. The prevalence in foxes has been around 40% in the last decade. Three trials of anthelmintic bait distribution have been conducted in Hokkaido to reduce the prevalence in foxes. In those trials, bait distribution was done along roads in the study area using cars and/or around fox breeding dens by hand. Changes in the prevalence in foxes were evaluated either by necropsy of captured foxes or by coproantigen and egg detection of faeces collected in field. All of the trials showed bait distribution was effective for the reduction of the prevalence in foxes; however, it was also suggested that a frequent and continuous baiting program is necessary for effective and stable control of the prevalence in foxes. As observed in some cities in Europe, urban foxes infected with the parasite were also recognized in Sapporo. A survey of pet dogs showed that 0.4% of surveyed dogs were determined infected. In addition, a dog which was transported from Hokkaido to the main island of Japan was found excreting E. multilocularis eggs. The results raised the public recognition of canine infections, which in turn lead to the modification of a Japanese law for infectious diseases and to the enforcement of a national reporting system of dogs infected with E. multilocularis by veterinarians. PMID- 16356763 TI - Role of chemotherapy of taeniasis in prevention of neurocysticercosis. AB - Neurocysticercosis (NC) is a serious public health problem in Taenia solium endemic areas and in some immigrants and international travelers. A key intervention in preventing NC is elimination of taeniasis by chemotherapy. Currently, two safe and effective taenicides, namely niclosamide and praziquantel, are available. Both are on WHO Essential Drug list, but are often inaccessible in T. solium endemic areas. Natural remedies, still widely used in some endemic areas, are frequently carcinogenic or highly toxic and as such should be discontinued. Chemotherapeutic intervention to control T. solium taeniasis/cysticercosis, whilst theoretically feasible, has several practical obstacles. These include poor public awareness, problems with diagnosing Taenia carriers, poor availability of taenicides where needed and low priority afforded to the control of NC. These can be overcome, respectively, by effective health education, wider use of newly developed coproantigen tests, strengthening of health services infrastructure and essential drugs distribution, and increasing the priority given to prevention of NC, as a leading cause of epilepsy in T. solium endemic areas. Information is accumulating on rational approaches to population-based short-term chemotherapeutic control measures. These are: widely available modern diagnostic tools and taenicides, treatment of any case of taeniasis, confirmed or probable, focus-oriented chemotherapy, irrespective of Taenia species implicated, improved sanitation, cooperation of veterinary and medical services, linkage with programs against epilepsy and cooperation of better educated communities. Now, it remains to take an advantage of existing tools and experience. PMID- 16356764 TI - Modelling and interpretation of gas detection using remote laser pointers. AB - We have developed a quantitative model of the performance of laser pointer style gas leak detectors, which are based on remote detection of backscattered radiation. The model incorporates instrumental noise limits, the reflectivity of the target background surface and a mathematical description of gas leak dispersion in constant wind speed and turbulence conditions. We have investigated optimum instrument performance and limits of detection in simulated leak detection situations. We predict that the optimum height for instruments is at eye level or above, giving an operating range of 10 m or more for most background surfaces, in wind speeds of up to 2.5 ms(-1). For ground based leak sources, we find laser pointer measurements are dominated by gas concentrations over a short distance close to the target surface, making their readings intuitive to end users in most cases. This finding is consistent with the results of field trials. PMID- 16356765 TI - Determination of trace silver by solid substrate-room temperature phosphorescence quenching method based on double catalytic system of meta-nitrophenyfluorone polyoxyethylene-chromium-potassium bromate-beta-cyclodextrin. AB - A new solid substrate-room temperature phosphorescence (SS-RTP) quenching method for the determination of trace silver has been established. It is based on the fact that when using Mg(2+) as ion perturber and beta-CD as surfactant, the system of meta-nitrophenyfluorone (R)-polyoxyethylene-Cr(III) can emit strong and stable room temperature phosphorescence signal on filter paper whose surface is modified by polyvinyl alcohol (PVA)-H(3)BO(3)-NaOH. Ag(I) can catalyze KBrO(3) oxidizing R-PEO-Cr(III) system which causes the quenching of SS-RTP. The reducing value of phosphorescence intensity (DeltaI(p)) is directly proportional to the concentration of Ag(I) in the range of 3.2-160 ag spot(-1) (corresponding concentration: 2.43 fg ml(-1), the sample volume: 0.40 microl spot(-1)) with a detection limit (LD) of 0.97 ag spot(-1). The regression equation of working curve can be expressed as DeltaI(p)=13.92+0.3089m(Ag)+ (ag spot(-1)) (r=0.9983, n=6). This method has many advantages, such as a wide linear dynamic range, high sensitivity, good repeatability and selectivity. It has been applied to the determination of trace silver in real samples with satisfactory results. What is more, the mechanism of SS-RTP quenching method based on Ag(I) catalyzing KBrO(3) oxidizing meta-nitrophenyfluorone has also been discussed. PMID- 16356766 TI - The ALPHA Project: an architecture for leveraging public health applications. AB - OBJECTIVE: Public health surveillance applications are central to the collection, analysis and dissemination of disease and health information. As these applications evolve and mature, it is evident that many of these applications must address similar requirements, such as policies, security and flexibility. It is important a software architecture is created to meet these requirements. METHODS: We outline the requirements for a public health surveillance application, and define a set of common components to address these requirements. These components are configured to produce services used in the development of public health applications. RESULTS: A layered software architecture, the ALPHA architecture, has been developed to support the development of public health applications. The architecture has been used to build eleven surveillance applications for the Public Health Agency of Canada in the areas of disease surveillance, survey, distributed data collection and inventory management. CONCLUSIONS: We have found that a software architecture that addresses requirements on policies, security and flexibility facilitates the development of configurable public health applications. By creating this architecture, key success factors, such as reducing cost and time-to-market of applications, adapting to changing surveillance targets and increasing user efficiency are achieved. PMID- 16356767 TI - Principal components analysis of Laplacian waveforms as a generic method for identifying ERP generator patterns: I. Evaluation with auditory oddball tasks. AB - OBJECTIVE: To evaluate the effectiveness and comparability of PCA-based simplifications of ERP waveforms versus their reference-free Laplacian transformations for separating task- and response-related ERP generator patterns during auditory oddball tasks. METHODS: Nose-referenced ERPs (31 sites total) were recorded from 66 right-handed adults during oddball tasks using syllables or tones. Response mode (left press, right press, silent count) and task was varied within subjects. Spherical spline current source density (CSD) waveforms were computed to sharpen ERP scalp topographies and eliminate volume-conducted contributions. ERP and CSD data were submitted to separate covariance-based, unrestricted temporal PCAs (Varimax) to disentangle temporally and spatially overlapping ERP and CSD components. RESULTS: Corresponding ERP and CSD factors were unambiguously related to known ERP components. For example, the dipolar organization of a central N1 was evident from factorized anterior sinks and posterior sources encompassing the Sylvian fissure. Factors associated with N2 were characterized by asymmetric frontolateral (tonal: frontotemporal R > L) and parietotemporal (phonetic: parietotemporal L > R) sinks for targets. A single ERP factor summarized parietal P3 activity, along with an anterior negativity. In contrast, two CSD factors peaking at 360 and 560 ms distinguished a parietal P3 source with an anterior sink from a centroparietal P3 source with a sharply localized Fz sink. A smaller parietal but larger left temporal P3 source was found for silent count compared to button press. Left or right press produced opposite, region-specific asymmetries originating from central sites, modulating the N2/P3 complex. CONCLUSIONS: CSD transformation is shown to be a valuable preprocessing step for PCA of ERP data, providing a unique, physiologically meaningful solution to the ubiquitous reference problem. By reducing ERP redundancy and producing sharper, simpler topographies, and without losing or distorting any effects of interest, the CSD-PCA solution replicated and extended previous task- and response-related findings. SIGNIFICANCE: Eliminating ambiguities of the recording reference, the combined CSD-PCA approach systematically bridges between montage-dependent scalp potentials and distinct, anatomically-relevant current generators, and shows promise as a comprehensive, generic strategy for ERP analysis. PMID- 16356768 TI - Principal components analysis of Laplacian waveforms as a generic method for identifying ERP generator patterns: II. Adequacy of low-density estimates. AB - OBJECTIVE: To evaluate the comparability of high- and low-density surface Laplacian estimates for determining ERP generator patterns of group data derived from a typical ERP sample size and paradigm. METHODS: High-density ERP data (129 sites) recorded from 17 adults during tonal and phonetic oddball tasks were converted to a 10-20-system EEG montage (31 sites) using spherical spline interpolations. Current source density (CSD) waveforms were computed from the high- and low-density, but otherwise identical, ERPs, and correlated at corresponding locations. CSD data were submitted to separate covariance-based, unrestricted temporal PCAs (Varimax of covariance loadings) to identify and effectively summarize temporally and spatially overlapping CSD components. Solutions were compared by correlating factor loadings and scores, and by plotting ANOVA F statistics derived from corresponding high- and low-resolution factor scores using representative sites. RESULTS: High- and low-density CSD waveforms, PCA solutions, and F statistics were remarkably similar, yielding correlations of .9 < or = r < or = .999 between waveforms, loadings, and scores for almost all comparisons at low-density locations except for low-signal CSD waveforms at occipital sites. Each of the first 10 high-density factors corresponded precisely to one factor of the first 10 low-density factors, with each 10-factor set accounting for the meaningful CSD variance (> 91.6%). CONCLUSIONS: Low-density surface Laplacian estimates were shown to be accurate approximations of high-density CSDs at these locations, which adequately and quite sufficiently summarized group data. Moreover, reasonable approximations of many high-density scalp locations were obtained for group data from interpolations of low-density data. If group findings are the primary objective, as typical for cognitive ERP research, low-resolution CSD topographies may be as efficient, given the effective spatial smoothing when averaging across subjects and/or conditions. SIGNIFICANCE: Conservative recommendations for restricting surface Laplacians to high-density recordings may not be appropriate for all ERP research applications, and should be re-evaluated considering objective, costs and benefits. PMID- 16356769 TI - MAP kinase kinase kinases and innate immunity. AB - Toll-like receptors, which respond to invariant microbial molecules, and receptors for the proinflammatory cytokines tumour necrosis factor and interleukin-1 are crucial for initiation and regulation of innate immune responses. These receptors activate each of the major mitogen-activated protein (MAP) kinase subtypes, extracellular signal-regulated protein kinases, c-Jun amino-terminal kinases and p38 MAP kinases, which are crucial for cell survival and controlling the expression of immune mediators. Here we discuss recent studies characterizing the specific MAP kinase kinase kinases (MAP 3-kinases) that link MAP kinases to receptors involved in innate immunity and the mechanisms by which the activity of MAP 3-kinases is regulated by such receptors. PMID- 16356770 TI - RANKL-RANK signaling in osteoclastogenesis and bone disease. AB - Hundreds of millions of people worldwide are affected by bone-related diseases, such as osteoporosis and rheumatoid arthritis. Understanding the molecular mechanisms of bone metabolism is crucial for developing novel drugs for treating such diseases. In particular, genetic experiments showing that the receptor activator of NF-kappaB (RANK), its ligand RANKL, and the decoy receptor OPG are essential, central regulators of osteoclast development and osteoclast function were significant turning points in our understanding of bone diseases. RANKL-RANK signaling activates a variety of downstream signaling pathways required for osteoclast development. Moreover, molecular cross-talk between RANKL-RANK and other ligand-receptor systems fine-tunes bone homeostasis in normal physiology and disease. Designing novel drugs that target RANKL-RANK and their signaling pathways in osteoclasts could potentially revolutionize the treatment of many diseases associated with bone loss such as arthritis, tooth loss, cancer metastases or osteoporosis. PMID- 16356772 TI - HPV vaccines. AB - The ability to generate human papillomavirus virus (HPV)-like particles by the synthesis and self-assembly in vitro of the major virus capsid protein L1 has transformed our prospects for preventing cervical carcinoma in women. These particles provide vaccines that are immunogenic and safe, and data from proof-of principle efficacy trials strongly suggest that they will protect against persistent HPV infection and cervical intraepithelial neoplasia. However, the duration of protection provided by these vaccines is not known, the antibody responses induced are HPV-type-specific and immunisation must occur pre-exposure to the virus. Second-generation vaccines could include an early antigen for protection post exposure and alternative delivery systems might be needed for the developing world. Therapeutic vaccines for low-grade intraepithelial disease are realistic but high-grade disease presents major hurdles for immunotherapies. PMID- 16356773 TI - Sleeping Beauty-mediated transposition and long-term expression in vivo: use of the LoxP/Cre recombinase system to distinguish transposition-specific expression. AB - The Sleeping Beauty transposon system (SB) has been shown to mediate nonviral integration of expression constructs resulting in long-term gene expression in several mammalian targets. Often, however, it is difficult to discern long-term expression resulting from transposition vs nonhomologous chromosomal recombination or maintenance of plasmid DNA in an extrachromosomal form. We have designed a system to silence expression from nontransposed sequences, making it possible to determine more specifically the amount of expression resulting from transposition. A transposon plasmid, pT2F/Cage (carrying a murine erythropoietin (Epo) gene transcriptionally regulated by the ubiquitously expressed CAGS promoter), was engineered to contain LoxP sites positioned so as to interrupt expression upon Cre-mediated recombination. Upon transposition these sites become segregated, thus protecting the expression construct from Cre-mediated recombination and subsequent silencing. Interferon-inducible Mx1Cre mice were administered pT2F/Cage with or without transposase-encoding plasmid. At 2 to 4 weeks postinjection, in the absence of SB transposase, Cre induction reduced Epo expression to about 1% of that seen in the group that was administered transposase-encoding plasmid, which maintained Epo levels near those of the uninduced groups. Southern hybridization analysis and plasmid rescue of transfected tissue supported the efficient Cre-mediated silencing of nontransposed sequences. These results indicate a substantial level of DNA mediated expression not associated with transposition, but which can be quantitatively distinguished from transposition by its sensitivity to Cre recombinase. The results also provide additional evidence for the effectiveness of the Sleeping Beauty transposon system as an in vivo DNA-mediated gene transfer strategy for achieving long-term expression. PMID- 16356774 TI - Aortopulmonary window with anomalous origin of the right coronary artery from the pulmonary artery: two cases highlighting the importance of complete pre-operative echocardiographic evaluation of the coronary arteries in all conotruncal anomalies. AB - This report describes two infants with an aortopulmonary window in association with anomalous origin of the right coronary artery from the pulmonary artery. In both cases the diagnosis was made pre-operatively by transthoracic echocardiography, with the initial clue being extensive collateral flow within the myocardium. In each case there was surgical confirmation of the echocardiographic findings. These two cases demonstrate that coronary artery evaluation should be an integral part of every new echocardiographic evaluation, particularly in the setting of conotruncal anomalies. PMID- 16356775 TI - Complex behavioral automatism arising from insular cortex. AB - We describe two cases of complex partial seizures with ictal violent movements arising from the insular cortex. The first patient, a 14-year-old girl, presented with hyperkinetic behavior such as rolling, thrashing, and pedaling, and the second case, a 38-year-old woman, had been suffering from frequent daytime hyperkinetic seizures characterized by bizarre vocalization, jumping, and violent bimanual movements. Both patients showed a slight high signal change in the right posterior ventral insular cortex in fluid-attenuated inversion recovery (FLAIR) studies involving magnetic resonance imaging, and extensive subdural electroencephalographic monitoring revealed EEG seizure onset from the temporal lobe. The posterior ventral insular and lateral temporal cortices were resected, resulting in complete seizure freedom in both cases. The histological diagnoses were focal cortical dysplasia in the first case and gliosis in the second case. There may exist a group of patients with complex partial seizures with ictal violent automatism that can be ameliorated by the resection of epileptogenic lesions in the insular cortex. Careful inspection of the insular cortex is necessary to diagnose this type of epileptic seizure. PMID- 16356776 TI - Epilepsy increases vulnerability of long-term face recognition to proactive interference. AB - Proactive interference (PI) decreases short- and long-term memory in healthy subjects. Neurological patients exhibit a heightened PI effect on short-term memory. It is, however, not known if PI affects long-term memory in neurological patients. We analyzed whether epilepsy heightens the negative effect of PI on long-term face memory. PI was induced by a list of 20 faces learned 24 hours prior to a target list of 20 faces. We tested immediate and 24-hour recognition for both lists. Twelve healthy controls and 42 patients with generalized epilepsy or temporal lobe epilepsy (TLE) were studied. PI led to a decrease in 24-hour recognition in patients with generalized epilepsy and TLE but not in controls. Thus, PI may cause long-term memory disturbances in epilepsy patients. PI was also associated with decreased short-term memory, but only in right TLE. This confirms the dominant role of the right temporal lobe in short-term face memory. PMID- 16356777 TI - Prefrontal disturbances as the sole manifestation of simple partial nonconvulsive status epilepticus. AB - We describe a case of frontal lobe epilepsy with rare nocturnal generalized tonic clonic seizures and repeated prolonged episodes of altered behavior lasting 1 to 2 days. The changes consisted of poor organizational strategies, impaired set shifting, emotional indifference, reduced motivation, and impairment of emotional decision making. Memory and consciousness were undisturbed, as she was able to follow her profession as a teacher. During the episode, the EEG was marked by right frontal rhythmical spikes and waves with spread to the homologous left region. Behavioral abnormalities and EEG changes were successfully treated with intravenous diazepam. The clinical representation can be ascribed to alterations of prefrontal-subcortical circuits, especially the anterior cingulate and orbitofrontal circuits. This unique case is classified as simple partial nonconvulsive status epilepticus with prefrontal disturbances as the sole manifestation. PMID- 16356778 TI - Kluver-Bucy syndrome after mycoplasmal bronchitis. AB - Kluver-Bucy syndrome is a rare neurobehavioral condition characterized by visual agnosia, excessive oral tendencies, hypermetamorphosis, placidity, altered sexual behavior, and changes in dietary habits. This description of a 14-year-old boy presenting with complete Kluver-Bucy syndrome after Mycoplasma pneumoniae bronchitis is the first such case report. MRI revealed left temporal horn dilation and asymmetry of both temporal lobes. We hypothesize that the pathophysiology of our case is immune-mediated damage by M. pneumoniae resulting in vasculopathy. PMID- 16356779 TI - Adolescent depression: description, causes, and interventions. AB - Depression is one of the most prevalent of the psychiatric disorders and is common among individuals with epilepsy. Depression often begins in adolescence. The present review focuses on adolescent depression. In particular, this review first summarizes the definition, description, and classification of adolescent depression. Next, potential causes of adolescent depression are reviewed from a vulnerability-stress perspective. This part of the review focuses on the role of stressors and how stressors interact with genetic, biological, cognitive, personality, and interpersonal vulnerabilities to predict adolescent depression. Last, clinical aspects of adolescent depression are reviewed, including treatment and prevention of depression and the relation to epileptic disorders in adolescence. In sum, a substantial percentage of youth with epilepsy and seizures exhibit depression, and many are not diagnosed or treated in a timely manner. The present review shows that there are valid, empirically based assessments, treatments, and preventions for depression in adolescence that hold promise for reducing the significant burden associated with depression. PMID- 16356780 TI - Right hemispheric language dominance in a right-handed male with a right frontal tumor shown by functional transcranial Doppler sonography. AB - BACKGROUND: A 38-year-old, right-handed man with late-onset right frontal epilepsy due to a ganglioglioma and atypical right hemispheric language dominance is described. METHODS: Language dominance was investigated with functional transcranial Doppler sonography (fTCD), and language localization with functional magnetic resonance imaging (fMRI). RESULTS: During a word generation task, fTCD showed atypical right hemispheric language dominance, which was confirmed by fMRI using a semantic word comparison and a word stem completion task. This information helped to guide the resective procedure, which left the patient seizure-free and did not induce new deficits. CONCLUSION: Functional TCD appears to be a useful and reliable screening tool for determining hemispheric language dominance, even in patients with atypical language representation. Functional MRI may be used to confirm fTCD results and further localize eloquent cortex. PMID- 16356781 TI - Possible precipitation of myoclonic seizures with oxcarbazepine. AB - Oxcarbazepine is an antiepileptic drug widely used to treat partial seizures in children and adults. We report two children who developed mycolonic seizures and had abnormal electroencephalograms when oxcarbazepine was introduced. Although it is possible that the emergence of myoclonic seizures and deterioration of the electroencephalograms were due to the natural course of the disorder, clinicians should be aware of the possible detrimental effects of oxcarbazepine in children with seizures. PMID- 16356782 TI - A psychosocial view of anxiety and depression in epilepsy. AB - The aim of this study was to study anxiety and depression in patients with epilepsy and evaluate their relationships with neuroepilepsy and psychological variables. neuroepilepsy and psychological variables. Sixty patients and 60 healthy subjects were interviewed at the outpatient clinic for epilepsy, using the Beck Depression Inventory and State-Trait-Anxiety Inventory. The objective of the semistructured interview was to identify the patients' perception of the disease, self-concept, personal strategies, and perception of seizure control. There was a significant difference in anxiety and depression between the groups, as well as a strong relationship between perception of seizure control and depression and anxiety, independently assessed. Epilepsy was associated with disease (63.4%), mental problems (11.6%), feelings of shame, fear, worry, and low self-esteem (56.6%), and perception of stigma (26.6%). The strategies were: looking for social support, seeking medical treatment, withdrawal, denial, and spiritual support. There was a significant association between psychological symptoms and perception of seizure control, which reinforces the importance of subjective aspects involved in epilepsy. PMID- 16356783 TI - Diversity of long-chain toxins in Tityus zulianus and Tityus discrepans venoms (Scorpiones, Buthidae): molecular, immunological, and mass spectral analyses. AB - In Venezuela, stings by Tityus zulianus scorpions produce cardiorespiratory arrest, whereas envenoming by Tityus discrepans involves gastrointestinal/pancreatic complications, suggesting structural and/or functional differences. We sought to compare their toxin repertoires through immunological, molecular, and mass spectral analyses. First, in vivo tests showed that neutralization of T. zulianus venom toxicity by the anti-T. discrepans antivenom was not complete. To compare T. discrepans and T. zulianus long-chain (sodium channel-active) toxins, their most toxic Sephadex G-50 fractions, TdII and TzII, were subjected to acid-urea PAGE, which showed differences in composition. Amplification of toxin-encoding mRNAs using a leader peptide-based oligonucleotide rendered cDNAs representing twelve T. discrepans and two T. zulianus distinct toxin transcripts, including only one shared component, indicating divergence between T. zulianus and T. discrepans 5' region-encoded, toxin signal peptides. A 3'-UTR polymorphism was also noticed among the transcripts encoding shared components Tz1 and Td4. MALDI-TOF MS profiling of TdII and TzII produced species-specific spectra, with seven of the individual masses matching those predicted by cDNA sequencing. Phylogenetic analysis showed that the unique T. zulianus transcript-encoded sequence, Tz2, is structurally related to Tityus serrulatus and Centruroides toxins. Together with previous reports, this work indicates that T. zulianus and T. discrepans toxin repertoires differ structurally and functionally. PMID- 16356784 TI - Pharmacogenetics of antihypertensive treatment. AB - Hypertension is a common disorder associated with increased cardiovascular morbidity and mortality. Unfortunately, in the US only about one-third of those who are aware of their hypertensive status have their blood pressure adequately controlled. One reason for this is the variable and unpredictable response individuals have to pharmacologic treatment. Clinicians often resort to "trial and-error" to match patients with effective drug treatment. Hypertension pharmacogenetics seeks to find genetic predictors of drug response. To date, more than forty studies have investigated associations between genetic polymorphisms and response to antihypertensive drugs. Angiotensin-converting enzyme inhibitors and beta blockers have been most frequently studied, followed by angiotensin II blockers, diuretics, adrenergic alpha-agonists, and calcium channel blockers. Renin-angiotensin-aldosterone system genes have been the most widely studied, with the angiotensin-converting enzyme I/D variant being typed in about one-half of all hypertension pharmacogenetic studies. In total, 160 possible gene polymorphism-drug interactions have been explored, with about one-quarter of these showing that genes predict drug response. However, disparate and conflicting findings have been the rule rather than the exception, and the discovery of clinically relevant antihypertensive drug-response genes remains elusive. While there is a growing enthusiasm that pharmacogenetics of hypertension is important, the translation of pharmacogenetic findings to clinical practice in the future will depend on additional studies to enhance our pharmacogenetics knowledge base, the availability of pharmacogenetic screening tests that are affordable and easy to implement in clinical practice, a cohort of clinicians who are trained to interpret genetic test results, and health care systems that pay for them. Caution regarding the future of hypertension pharmacogenetics is warranted. PMID- 16356785 TI - Expression pattern of the maternally imprinted gene Gtl2 in the forebrain during embryonic development and adulthood. AB - Recent work has uncovered a large number of imprinted genes, many of which are thought to play a role in neurodevelopment and behavior. In order to begin to understand the role of specific genes in these processes, their expression patterns will be key. In this study we used in situ hybridization to study the developmental expression of Gtl2 in the forebrain from E12.5 to adulthood, since preliminary data from a microarray study indicated differential expression between the ventral and dorsal telencephalon of the mouse at a critical time point in the generation and migration of cortical neuronal populations. Strong expression was observed in the diencephalon, ventral telencephalon, post mitotic cell layers of the neocortex and pyramidal cell layer of the hippocampus. Additionally, heavily labeled subpopulations of laminar restricted cells were seen in the latter two areas. PMID- 16356786 TI - Remote NADH imaging through an ordered array of electrochemiluminescent nanoapertures. AB - In this report, we present an ordered array comprising thousands of nanoapertures for the electrochemiluminescent (ECL) detection of NADH. It was fabricated on the distal face of a coherent optical fiber bundle. Such a high-density array of nanoapertures combines optical, imaging and electrochemical properties. Indeed, each nanoaperture is surrounded by a gold nanoring, which acts as an electrode material. The behavior of the array was characterized by cyclic voltammetry and it shows excellent electrochemical performances. NADH is the analyte, which is measured in presence of Ru(bpy)3(2+). The ruthenium complex mediates the NADH oxidation and this coenzyme acts as a co-reactant in the ECL mechanism. ECL light is generated at the distal face of the array by each gold ring electrode. A fraction of this ECL light is collected by the corresponding nanoaperture, transmitted through the optical fiber bundle and finally imaged on the proximal face with a CCD camera. In this work, we show that NADH concentration is remotely detected by an oxidative-reductive ECL mechanism. We present also some preliminary results about the ECL process of NADH with Ru(bpy)3(2+). The ECL behavior of NADH on gold surface is reported. The influence of the applied potential on the collected light intensity was investigated. The variation of the ECL intensity measured through the nanoaperture array with NADH concentration is linear. Remote ECL detection of NADH is spatially resolved over a large area with a micrometer resolution through the array. Therefore, such array integrates several complementary functions: ECL light generation, collection, transmission and remote imaging in an array format. PMID- 16356788 TI - Simultaneous measurement of urinary polyols using gas chromatography/mass spectrometry. AB - In the present study, we simultaneously measured several polyols, such as adonitol, arabitol, dulcitol, glucose, myo-inositol, mannitol, sorbitol, and xylitol, in urine by gas chromatography/mass spectrometry-positive chemical ionization. We also examined possible relationship between the levels of these metabolites and age in normal individuals. In order to proceed to its quantification by GC/MS, 200 microL of a urine sample were diluted with 3 ml of distilled water, lyophilized, acetylated, and then analyzed them. Using this method, we were able to quantify as little as 0.5-1.0 ng/microL, and we made the calibration curves to be linear from 0.25 to 250 ng/microL (r(2)>0.991). Analytical recoveries were over 89.4%, and the inter-day and intra-day variability for accuracy and reproducibility was less than 20%. In the normal urine sample, the levels of polyols were gender-differentiated and age-related. This simple GC/MS method is sensitive and allows the measurement of wide ranges of polyols using small amounts of urine. We conclude that the quantitation of urinary polyols using GC/MS appears to be a clinically useful method for assessing polyol-pathway activity. PMID- 16356787 TI - Attenuation of host defense function of lung phagocytes in young cystic fibrosis patients. AB - BACKGROUND: Recent reports suggest that endotoxin exposure can blunt phagocyte functions. The aim of this study was to examine whether lung phagocytic cells have altered host defense function in young cystic fibrosis (CF) patients, and to explore the contribution of neutrophil elastase (NE) and surfactant proteins to these effects. METHODS: BALF cells from CF children (N=12) and disease controls (N=12) were analyzed by flow cytometry for mCD14 and HLA-DR expression and phagocytosis. The effects of exogenous surfactant protein A and D (SP-A,D) and proteases on BALF cells in short term culture were assessed experimentally. RESULTS: Expression of the surface markers mCD14 and HLA-DR, and phagocytosis, were all blunted on CF phagocytes compared to disease controls (p<0.05). In CF phagocytes, SP-A enhanced both phagocytosis and mCD14 expression (p<0.05). Both CF BALF and NE reduced phagocytosis and expression of mCD14 and HLA-DR (p<0.05) by non-CF phagocytes; the latter effect was attenuated by protease inhibitor. CONCLUSION: CF airway phagocytes appear to have altered host defense functions that could contribute to poor bacterial clearance. These impairments can be reproduced by incubation of non-CF cells with NE, while SP-A can partially reverse them. Decreasing protease activity and increasing collectin activity may be beneficial in early CF. PMID- 16356789 TI - Comparison of liquid chromatography with fluorescence detection to liquid chromatography-mass spectrometry for the determination of fluoxetine and norfluoxetine in human plasma. AB - A comparison study on fluoxetine (FL) and norfluoxetine (NORFL) quantitation in human plasma was carried out between the recently developed liquid chromatographic method with fluorescence detection (LC-FLD) and an earlier established liquid chromatography-mass spectrometry (LC-MS) laboratory procedure. Comparative method evaluation was based on the analysis of plasma samples obtained from Parkinsonian patients receiving 20mg of FL per day. The LC-FLD method involves a two-step liquid extraction procedure without any derivatization, followed by direct chromatography on a Zorbax C8 reversed-phase column. The analytical results are discussed in terms of the method validation and the corresponding experimental protocol (r>/=0.998; CV<9%; LOQ 20 microg/l). There was good correlation between FL, as well as NORFL, plasma levels as determined by the LC-MS and LC-FLD techniques (r=0.9597, N=16 and r=0.9852, N=14 for FL and NORFL, respectively). The results confirm that direct FL/NORFL fluorimetric determination is acceptable for routine use in pharmacokinetic and clinical studies. PMID- 16356790 TI - Simultaneous determination of four active alkaloids from a traditional Chinese medicine Corydalis saxicola Bunting. (Yanhuanglian) in plasma and urine samples by LC-MS-MS. AB - A sensitive rapid method for the simultaneous determination of four major active alkaloids (dehydrocavidine, coptisine, dehydroapocavidine, and tetradehydroscoulerine, in abbreviation thereafter called YHL-I, YHL-II, YHL-III, and YHL-IV, respectively) from a Chinese traditional medicine Corydalis saxicola Bunting. (Yanhuanglian) in rat plasma and urine was established and validated. The assay for these substances in plasma and urine was based on HPLC coupled with tandem mass spectrometry (MS/MS) detection using multiple reaction monitoring mode (MRM) with berberine and clenbuterol as internal standards. The plasma and urine sample were deproteinated by adding methanol prior to liquid chromatography where separation was performed on a Luna column (5 microm, 100 x 2.00 mm) and an Agilent Zorbax SB-C18 guard column (5 microm, 20 x 4 mm). The method was validated with the concentration range 1-1000 ng/mL in plasma and 10-1000 ng/mL in urine for the four test compounds, and the calibration curves were linear with correlation coefficients >0.999. The lowest limits of quantitation for all four substances were 1 ng/mL in 0.1 mL rat plasma and 10 ng/mL in 0.1 mL urine. The intra-assay accuracy and precision in plasma ranged from 88.1 to 115.7% and 1.4 to 10.8%, respectively, while inter-assay accuracy and precision for YHL-I, YHL II, YHL-III, and YHL-IV ranged from 96.2 to 113.2% and 0.4 to 16.9%, respectively. The intra-assay accuracy and precision for YHL-I, YHL-II, YHL-III, and YHL-IV in rat urine ranged from 96.1 to 112.9% and 1.2 to 8.3%, respectively, while inter-assay accuracy and precision ranged from 95.0 to 106.8% and 2.2 to 10.3%, respectively. The method was further applied to assess pharmacokinetics and urine excretion of the four alkaloids after oral and intravenous administration to rats. Practical utility of this new LC-MS-MS method was confirmed in pilot pharmacokinetic studies in rats following both intravenous and oral administration. PMID- 16356791 TI - A simple and fast extraction method for organochlorine pesticides and polychlorinated biphenyls in small volumes of avian serum. AB - A solid-phase extraction (SPE) method was developed using 8M urea to desorb and extract organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs) from avian serum for analysis by capillary gas chromatography with electron capture detection (GC-ECD). The analytes were efficiently extracted from the denatured serum-lipoprotein-analyte complex by one passage through an Oasis((R)) hydrophilic-lipophilic-balanced (HLB) SPE cartridge. No further clean-up was necessary, the entire extraction procedure and GC-ECD analysis can be accomplished in less than 3h. Serum volumes ranged from 100 microL to 1 mL with absolute recoveries of 90-101% for PCBs and 74% to 101% for the OC pesticides. PMID- 16356793 TI - Update of renal imaging. AB - Significant technical improvements have allowed the use of radiological techniques to play a growing role in the imaging of renal diseases. Noninvasive ultrasound methods (ie, sonography and Doppler) are now positioned as first-line methods for the evaluation of renovascular diseases. Multidetector computed tomography is able to provide high spatial resolution images of the kidneys and renal arterial vessels. Magnetic resonance imaging, which provides higher signal to-noise ratio and higher spatial and/or temporal resolution, can display both morphological information about renal parenchyma and vessels and functional data, including perfusion, filtration, diffusion, or oxygenation. In renovascular diseases, these techniques have the potential to drive new strategies, including Doppler sonography as a first-line method, followed by computed tomography angiography or magnetic resonance angiography, depending mainly on renal function. Imaging of parenchymal renal diseases is developing toward more quantitative (volumetric and functional measurements) and more specific (through in vivo cell targeting) acquisitions for obtaining the adequate information on tissue characteristics relevant either for diagnosis or for prognosis or treatment follow-up. PMID- 16356794 TI - Pediatric applications of renal nuclear medicine. AB - This review should be regarded as an opinion based on personal experience, clinical and experimental studies, and many discussions with colleagues. It covers the main radionuclide procedures for nephro-urological diseases in children. Glomerular filtration rate can be accurately determined using simplified 2- or 1-blood sample plasma clearance methods. Minor controversies related to the technical aspects of these methods concern principally some correction factors, the quality control, and the normal values in children. However, the main problem is the reluctance of the clinician to apply these methods, despite the accuracy and precision that are higher than with the traditional chemical methods. Interesting indications are early detection of renal impairment, hyperfiltration status, and monitoring of nephrotoxic drugs. Cortical scintigraphy is accepted as a highly sensitive technique for the detection of regional lesions. It accurately reflects the histological changes, and the interobserver reproducibility in reporting is high. Potential technical pitfalls should be recognized, such as the normal variants and the difficulty in differentiating acute lesions from permanent ones or acquired lesions from congenital ones. Although dimercaptosuccinic acid scintigraphy seems to play a minor role in the traditional approach to urinary tract infection, recent studies suggest that this examination might influence the treatment of the acute phase, the indication for chemoprophylaxis and micturating cystography, and the duration of follow-up. New technical developments have been applied recently to the renogram: tracers more appropriate to the young child, early injection of furosemide, late postmicturition and gravity-assisted images and, finally, more objective parameters of renal drainage. Pitfalls mainly are related to the interpretation of drainage on images and curves. Dilated uropathies represent the main indication of the renogram, but the impact of this technique on the management of the child is, in a great number of cases, still a matter of intense controversy. Direct and indirect radionuclide cystography are interesting alternatives to the radiograph technique and should be integrated into the process of diagnosis and follow-up of vesicoureteral reflux. PMID- 16356796 TI - Nuclear medicine studies of the prostate, testes, and bladder. AB - During the last decade, there has been a significant advancement in imaging of urologic diseases. Transrectal ultrasound (TRUS), computerized tomography (CT), magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS), and positron emission tomography (PET) are still experiencing new developments in urology. Despite these many technological advances, the initial diagnostic procedure for a patient with suspected prostate cancer (PC) is multiple site blind prostate biopsies. There is a need for a noninvasive metabolic imaging modality to direct the site of biopsy to decrease the sampling error. MRS seems promising but as it is a costly and more time-consuming test, further studies are needed to evaluate its clinical utility. Currently, PET does not play any role to direct biopsy. Acetate and choline appear to be better tracers than FDG for the detection of a prostate lesion, however, further well-organized studies are needed before any of these agents can be used clinically. Incidental detection of intense focal uptake in the prostate during whole body PET scanning should be evaluated with prostate-specific antigen (PSA) and TRUS-guided biopsy. Although FDG is inferior to other tracers for primary staging, it may be useful in selected patients with suspected high-grade cancer. The role of ProstaScint scan is still controversial for detection of recurrent PC. This study may be helpful for evaluating nodal metastases when PSA is elevated and bone scan is negative. Bone scan remains the study of choice when bone metastases are suspected (PSA>15 20 ng/mL+/-bone pain). Acetate and choline provide better accuracy than FDG in the detection of local soft tissue disease, nodal involvement, and distant metastases. High FDG uptake may be indicative of more aggressive and possibly androgen-independent disease. PET/CT with any of the above PET tracers will most likely be preferred to the PET scan alone due to better localization of a hot lesion in PET/CT. Nuclear medicine studies also have been used to evaluate acute scrotum and testicular neoplasms. Scrotal scintigraphy has lost its popularity to Doppler ultrasound in the evaluation of the acute scrotum. In testicular tumors, FDG-PET appears to be superior to conventional imaging modalities in initial staging, detection of residual/recurrence, and monitoring treatment response. Tumor markers after treatment occasionally are elevated and cannot locate the site of recurrence, FDG-PET can play a very important role in this regard. Nuclear medicine studies also have been used to evaluate diseases of the urinary bladder. Radionuclide cystography is more sensitive and has less than 1/20 the radiation exposure of the conventional contrast enhanced micturating cystourethrogram (MCU). However, the utility of FDG-PET in the evaluation of bladder cancer seems to be limited to the evaluation of distant metastases. 11C Methionine and choline may be a better option for local and nodal disease due to their negligible excretion in the urine. PMID- 16356797 TI - The role of fluorodeoxyglucose, 18F-dihydroxyphenylalanine, 18F-choline, and 18F fluoride in bone imaging with emphasis on prostate and breast. AB - Diagnostic imaging has played a major role in the evaluation of patients with bone metastases. The imaging modalities have included bone scintigraphy, computed tomography, magnetic resonance imaging, and most recently PET/CT, which can be performed with different tracers, including fluorodeoxyglucose (FDG), 18F fluoride, 18F-choline (FCH), and 18F-DOPA (dihydroxyphenylalanine). For most tumors the sensitivity of FDG in detecting bone metastases is similar to bone scintigraphy; additionally it can be used to monitor the response to chemotherapy and hormonal therapy. 18F-Fluoride may provide a more sensitive "conventional" bone scan and is superior for FDG nonavid tumors, but, nevertheless, FDG in "early disease" often has clear advantages over 18F-fluoride. Although more data need to be obtained, it appears that FCH is highly efficient in preoperative management regarding N and M staging of prostate cancer once metastatic disease is strongly suspected or documented. For neuroendocrine tumors and in particular in medullary thyroid cancer, DOPA is similar to 18F-fluoride in providing high quality information regarding the skeleton. Nevertheless, prospective studies with large patient groups will be essential to define the exact diagnostic role of FCH and DOPA PET in different clinical settings. PMID- 16356795 TI - Future direction of renal positron emission tomography. AB - Positron emission tomography (PET) is perfectly suited for quantitative imaging of the kidneys, and the recent improvements in detector technology, computer hardware, and image processing software add to its appeal. Multiple positron emitting radioisotopes can be used for renal imaging. Some, including carbon-11, nitrogen-13, and oxygen-15, can be used at institutions with an on-site cyclotron. Other radioisotopes that may be even more useful in a clinical setting are those that either can be obtained from radionuclide generators (rubidium-82, copper-62) or have a sufficiently long half-life for transportation (fluorine 18). The clinical use of functional renal PET studies (blood flow, glomerular filtration rate) has been slow, in part because of the success of concurrent technologies, including single-photon emission computed tomography (SPECT) and planar gamma camera imaging. Renal blood flow studies can be performed with O-15 labeled water, N-13-labeled ammonia, rubidium-82, and copper-labeled PTSM. With these tracers, renal blood flow can be quantified using a modified microsphere kinetic model. Glomerular filtration can be imaged and quantified with gallium-68 EDTA or cobalt-55 EDTA. Measurements of renal blood flow with PET have potential applications in renovascular disease, in transplant rejection or acute tubular necrosis, in drug-induced nephropathies, ureteral obstruction, before and after revascularization, and before and after the placement of ureteral stents. The most important clinical application for imaging glomerular function with PET would be renovascular hypertension. Molecular imaging of the kidneys with PET is rather limited. At present, research is focused on the investigation of metabolism (acetate), membrane transporters (organic cation and anion transporters, pepT1 and pepT2, GLUT, SGLT), enzymes (ACE), and receptors (AT1R). Because many nephrological and urological disorders are initiated at the molecular and organelle levels and may remain localized at their origin for an extended period of time, new disease-specific molecular probes for PET studies of the kidneys need to be developed. Future applications of molecular renal imaging are likely to involve studies of tissue hypoxia and apoptosis in renovascular renal disease, renal cancer, and obstructive nephropathy, monitoring the molecular signatures of atherosclerotic plaques, measuring endothelial dysfunction and response to balloon revascularization and restenosis, molecular assessment of the nephrotoxic effects of cyclosporine, anticancer drugs, and radiation therapy. New radioligands will enhance the staging and follow-up of renal and prostate cancer. Methods will be developed for investigation of the kinetics of drug-delivery systems and delivery and deposition of prodrugs, reporter gene technology, delivery of gene therapy (nuclear and mitochondrial), assessment of the delivery of cellular, viral, and nonviral vectors (liposomes, polycations, fusion proteins, electroporation, hematopoietic stems cells). Of particular importance will be investigations of stem cell kinetics, including local presence, bloodborne migration, activation, seeding, and its role in renal remodeling (psychological, pathological, and therapy induced). Methods also could be established for investigating the role of receptors and oncoproteins in cellular proliferation, apoptosis, tubular atrophy, and interstitial fibrosis; monitoring ras gene targeting in kidney diseases, assessing cell therapy devices (bioartificial filters, renal tubule assist devices, and bioarticial kidneys), and targeting of signal transduction moleculas with growth factors and cytokines. These potential new approaches are, at best, in an experimental stage, and more research will be needed for their implementation. PMID- 16356798 TI - Positron emission tomography in gynecologic cancer. AB - Most positron emission tomography (PET) imaging studies in gynecologic cancer are performed using (18)F-fluorodeoxyglucose (FDG). It contributes valuable information in primary staging of untreated advanced cervical cancer, in the post treatment surveillance with unexplained tumor marker (such as squamous cell carcinoma antigen [SCC-Ag]) elevation or suspicious of recurrence, and restaging of potentially curable recurrent cervical cancer. Its value in early-stage resectable cervical cancer is questionable. In ovarian cancer, FDG-PET provides benefits for those with plateaued or increasing abnormal serum CA 125 (>35 U/mL), computed tomography and/or magnetic resonance imaging (CT-MRI) defined localized recurrence feasible for local destructive procedures (such as surgery, radiotherapy, or radiofrequency ablation), and clinically suspected recurrent or persistent cancer for which CT-guide biopsy cannot be performed. The role of FDG PET in endometrial cancer is relatively less defined because of the lack of data in the literature. In our prospective study, FDG-PET coupled with MRI-CT may facilitate optimal management of endometrial cancer in well-selected cases. The clinical impact was positive in 29 (48.3%) of the 60 scans, 22.2% for primary staging, 73.1% for post-therapy surveillance, and 57.1% after salvage therapy, respectively. Scant studies have been reported in the management of vulvar cancer using FDG-PET. More data are needed. Gestational trophoblastic neoplasia is quite unique in biological behavior and clinical management. Our preliminary results suggest that FDG-PET is potentially useful in selected gestational trophoblastic neoplasia by providing a precise metastatic mapping of tumor extent up front, monitoring response, and localizing viable tumors after chemotherapy. The evaluation of a diagnostic tool, such as PET, is usually via comparing the diagnostic efficacy (sensitivity, specificity, etc), by using a more sophisticated receiver operating curve method, or the proportion of treatment been modified. Evaluating PET by clinical benefit is specific to the individual tumor and an attractive new endpoint. PMID- 16356799 TI - Recurrence of lymphoma presenting as asymmetrically increased testicular activity on FDG-PET/CT. PMID- 16356800 TI - Statin Therapy: Clinical Trials, Guidelines, and New Directions. Proceedings from the 3rd Atorvastatin Global Investigators Meeting in Toronto, Ontario, Canada, June 2004. PMID- 16356801 TI - Lipid-lowering therapy and the patient with multiple risk factors: what have we learned from the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT)? AB - The Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT) was the first trial of 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) to assess the benefits of lipid lowering in the primary prevention of coronary heart disease (CHD) in patients with hypertension who were not deemed to have dyslipidemia by conventional measures. A total of 19,342 patients with hypertension and > or =3 cardiovascular risk factors, but without CHD, were enrolled in ASCOT. Of these, 10,305 patients with a serum cholesterol level of < or =250 mg/dL (< or =6.5 mmol/L) were randomized to either atorvastatin (10 mg/day) or placebo in the ASCOT lipid-lowering arm (ASCOT-LLA). Follow-up was planned for an average of 5 years. The ASCOT-LLA was stopped after 3.3 years owing to the superiority of atorvastatin 10 mg over placebo in reducing the primary end point of nonfatal myocardial infarction (MI) and fatal CHD. Patients receiving atorvastatin experienced a significant reduction in total cholesterol (50 mg/dL [1.3 mmol/L]) and low-density lipoprotein cholesterol (46 mg/dL [1.2 mmol/L]) levels after 1 year compared with those who received placebo. Cholesterol lowering with atorvastatin was associated with a highly significant reduction in the primary end point of nonfatal MI and fatal CHD (36%, P = 0.0005). The observed benefit was consistent across the secondary end points and the 18 prespecified subgroups. The ASCOT-LLA findings have influenced lipid lowering guidelines and support the concept that treatment strategies to reduce cardiovascular disease should be based on the assessment of all cardiovascular risk factors, rather than on numerical thresholds of individual risk factors, to determine treatment strategies. PMID- 16356802 TI - Benefits of lipid-lowering therapy in patients with type 2 diabetes mellitus. AB - The incidence of type 2 diabetes mellitus is expected to increase dramatically over the next decade. Patients with type 2 diabetes are at a much greater risk for cardiovascular disease (CVD) than are nondiabetic individuals. Consequently, the treatment of CVD risk factors is a healthcare priority in this patient population. Dyslipidemia is a major cardiovascular (CV) risk factor in patients with type 2 diabetes, and it is characterized by elevated triglyceride levels, low high-density lipoprotein (HDL) cholesterol levels, and a preponderance of small, dense low-density lipoprotein (LDL) particles. Subgroup analyses of clinical trial data suggest that treatment of the entire range of lipid abnormalities may reduce CV risk in this patient population. The 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are the best therapy for LDL cholesterol reduction. A number of statin trials have shown significant CV risk reduction through LDL cholesterol lowering in subgroups of patients with diabetes. The recently published Collaborative Atorvastatin Diabetes Study (CARDS), a placebo-controlled trial conducted solely in patients with type 2 diabetes, terminated 2 years earlier than its anticipated length owing to the significant reduction in number of CV events observed in patients randomized to receive low-dose atorvastatin versus placebo. These results suggest that low-dose statin therapy with atorvastatin results in significant reduction of CV events in patients with type 2 diabetes without prior CVD or high LDL cholesterol levels. Based on this evidence, patients with type 2 diabetes may be candidates for statin therapy regardless of LDL cholesterol level and in the absence of a previous CV event. PMID- 16356803 TI - Statin use in a "real-world" clinical setting: aggressive lipid lowering compared with usual care in the Aggressive Lipid-Lowering Initiation Abates New Cardiac Events (ALLIANCE) trial. AB - Clinical trials that incorporate elements of "real-world" experience are of great value to practicing physicians. Using a trial design adapted to approximate clinical settings, the Aggressive Lipid-Lowering Initiation Abates New Cardiac Events (ALLIANCE) trial compared a focused treatment strategy using atorvastatin with usual medical care. Patients eligible for study participation were predefined based on diagnosis codes for coronary heart disease from US managed care database records; 66% of these patients were taking lipid-lowering medications at study entry. In contrast to standard clinical trials, ALLIANCE maintained a real-world environment by limiting the interactions of investigators with patients after dose titration of aggressive treatment to a low-density lipoprotein (LDL) cholesterol goal < 80 mg/dL (2.1 mmol/L) or maximum atorvastatin dose of 80 mg/day. After 51.5 months of follow-up, the study showed that aggressive treatment with atorvastatin was associated with significantly lower LDL cholesterol levels (147 mg/dL [3.8 mmol/L] to 95 mg/dL [2.5 mmol/L]) over usual care (146 mg/dL [3.8 mmol/L] to 111 mg/dL [2.9 mmol/L]). This greater reduction in LDL cholesterol was accompanied by improved outcomes in the composite primary end point of cardiovascular events (-17% with atorvastatin vs. usual care; P = 0.02) and particularly in the end point of nonfatal myocardial infarction (-47% with atorvastatin vs. usual care; P = 0.0002). No safety difference was noted between the 2 treatment groups. These results indicate that usual-care treatment was not equivalent to targeted statin therapy, even in a trial conducted to minimize potential bias owing to traditional patient selection and trial design methods. PMID- 16356804 TI - Halting the progression of atherosclerosis with intensive lipid lowering: results from the Reversal of Atherosclerosis with Aggressive Lipid Lowering (REVERSAL) trial. AB - Intravascular ultrasonography is a catheter-based technique used to provide 3 dimensional views of the vessel lumen as well as the size and distribution of atherosclerotic plaques. This imaging technique was used in the Reversal of Atherosclerosis with Aggressive Lipid Lowering (REVERSAL) study, an 18-month, randomized, controlled, multicenter trial comparing the effects of intensive versus moderate lipid-lowering therapy on plaque progression in patients requiring coronary angiography. A total of 253 patients were randomized to atorvastatin 80 mg/day (intensive lipid lowering) and 249 patients were randomized to pravastatin 40 mg/day (moderate lipid lowering). Low-density lipoprotein (LDL) cholesterol levels decreased from a baseline mean of 150 mg/dL (3.9 mmol/L) in both groups to 79 mg/dL (2.0 mmol/L) in the atorvastatin group and 110 mg/dL (2.9 mmol/L) in the pravastatin group. High-sensitivity C-reactive protein (hs-CRP) levels decreased by 36.4% in the atorvastatin group versus 5.2% in the pravastatin group (P <0.001). For the primary end point of percent change in total atheroma volume, a significantly lower rate of progression from baseline was observed with atorvastatin (-0.4%) than with pravastatin (2.7%) (P = 0.02). Linear regression analysis showed an inverse relation between lipid reduction and plaque progression for both groups; however, at any given level of LDL cholesterol, the progression rate was lower with atorvastatin compared with pravastatin. Both regimens were well tolerated. The results show that intensive lipid lowering with atorvastatin 80 mg/day for 18 months halted the progression of coronary atherosclerosis, whereas more moderate lipid lowering with pravastatin 40 mg/day was associated with progression. The differences in the progression rate are likely to be a result of greater reduction in atherogenic lipoproteins and hs-CRP with intensive therapy. PMID- 16356805 TI - Improved outcome after acute coronary syndromes with an intensive versus standard lipid-lowering regimen: results from the Pravastatin or Atorvastatin Evaluation and Infection Therapy-Thrombolysis in Myocardial Infarction 22 (PROVE IT-TIMI 22) trial. AB - The aim of the Pravastatin or Atorvastatin Evaluation and Infection Therapy Thrombolysis in Myocardial Infarction 22 (PROVE IT-TIMI 22) trial was to determine whether intensive low-density lipoprotein (LDL)-cholesterol lowering to a level of approximately 70 mg/dL (1.8 mmol/L) with atorvastatin 80 mg/day was more efficacious than standard LDL cholesterol lowering to 100 mg/dL (2.6 mmol/L) with pravastatin 40 mg/day in reducing the incidence of cardiovascular events in patients with acute coronary syndrome (ACS). In total, 4,162 men and women aged >18 years, who had been hospitalized for an ACS within the preceding 10 days, were randomized to receive either pravastatin 40 mg/day or atorvastatin 80 mg/day. The median LDL cholesterol levels achieved during follow-up were 95 mg/dL (2.5 mmol/L) in the pravastatin group and 62 mg/dL (1.6 mmol/L) in the atorvastatin group (P <0.001). Standard treatment (statin) with a 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor (pravastatin 40 mg/day) resulted in a 22% reduction in LDL cholesterol levels at 30 days compared with a 51% reduction with intensive therapy (atorvastatin 80 mg/day). At 2 years, a relative risk reduction of 16% (95% confidence interval, 5%-26%; P = 0.005) in the primary end point rate (death, myocardial infarction, documented unstable angina requiring hospitalization, coronary revascularization, or stroke) was seen in patients receiving intensive statin treatment compared with standard statin therapy. The benefit of intensive treatment was apparent as early as 30 days and was consistent over time. The PROVE IT-TIMI 22 data indicate that patients recently hospitalized for an ACS benefit from early and continued lowering of LDL cholesterol to levels substantially below current guideline recommendations. PMID- 16356806 TI - Barriers to effective implementation of guideline recommendations. AB - Cardiovascular disease (CVD) is the leading cause of death worldwide, and its prevention and treatment are important healthcare aims. Hypercholesterolemia is among the most important modifiable risk factors for CVD, and numerous guidelines exist for the treatment of this condition. Nevertheless, despite the existence of well-established and safe pharmacologic therapy for lowering cholesterol and preventing CVD, surveys in the United States and Europe have revealed that many patients have elevated cholesterol levels. There is a clear gap between what is known about treating CVD and the implementation of that knowledge. A survey assessing patients' knowledge about CVD observed that many patients are unaware of the disease prevalence and have little knowledge about the main risk factors, including the importance of cholesterol. Another survey demonstrated that many physicians overestimate patients' awareness of CVD and that physicians also overestimate the extent to which guidelines are implemented in clinical practice. Guideline implementation may be improved by narrowing the discrepancies between what patients and physicians believe and the reality. Many physicians claim that lack of time hinders guideline implementation and improvement of patient education. Physicians also appear to lack the motivation to implement lipid lowering interventions. A multifactorial approach to improving use of guidelines in clinical practice may improve the treatment and prevention of CVD. PMID- 16356807 TI - What impact will current trial data have on future guideline recommendations? AB - The current Third Joint European Societies' Guidelines on Cardiovascular Disease Prevention in Clinical Practice reflect an active approach to cardiovascular disease (CVD) prevention. These guidelines have shifted the emphasis from coronary heart disease to total atherosclerotic CVD. A new risk prediction system, Systematic Coronary Risk Evaluation (SCORE), has been developed to define risk in terms of absolute 10-year risk of a fatal cardiovascular event. The definition of high risk has also been refined. The currently recommended goals for total and low-density lipoprotein (LDL) cholesterol are based on the evidence pool that was available at the time of publication. More recent evidence from the Heart Protection Study (HPS), the lipid-lowering arm of the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT-LLA), and the Pravastatin or Atorvastatin Evaluation and Infection Therapy-Thrombolysis in Myocardial Infarction 22 (PROVE IT-TIMI 22) trial indicates that lowering LDL cholesterol levels beyond the currently recommended goals can produce incremental reductions in cardiovascular morbidity and mortality. Results from the recently completed Treating to New Targets (TNT) study lend further support to implementing lower lipid goals than those currently suggested by guidelines. Results from the ongoing Incremental Decrease in Endpoints through Aggressive Lipid Lowering (IDEAL) trial may provide additional evidence in support of the implementation of lower lipid goals. Nevertheless, the scope of future guidelines extends well beyond lipid targets. Guidelines of the future are likely to provide increasingly comprehensive recommendations on combined risk reduction, to publicize the characteristics of healthy people by highlighting acceptable levels of all cardiovascular risk factors, and to refine the concept of evidence-based medicine by shifting the focus from drug therapy to preventive therapy. PMID- 16356808 TI - Statin therapy in the treatment of Alzheimer disease: what is the rationale? AB - Alzheimer disease (AD) is a chronic neurodegenerative disorder that is manifested by cognitive decline, neuropsychiatric symptoms, and diffuse structural abnormalities in the brain. Its prevalence is predicted to rise 4-fold in the next 50 years. AD is characterized pathologically by deposition of extracellular beta-amyloid and accumulation of neurofibrillary tangles. Neuronal death and specific neurotransmitter deficits also are part of the pathologic picture. Strategies to delay symptom progression have focused on addressing the neurotransmitter deficits. Strategies to delay the onset or biologic progression of AD largely have targeted the plaques formed by the deposition of beta-amyloid. AD and cardiovascular disease share common risk factors, notably hypercholesterolemia, and occur together more often than expected by chance. Therapy with the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) is the first-line treatment option for hypercholesterolemia, and observational studies have suggested that the risk of AD is reduced in patients who receive statin therapy in midlife. This reduction in risk of AD observed with statin therapy may be due to statins reducing beta-amyloid formation and deposition or to their known anti-inflammatory effects. Two randomized double-blind statin trials in patients with AD to assess the potential for statins to slow disease progression are currently under way. If successful, statin AD primary prevention trials may be developed. PMID- 16356809 TI - A rationale for combination therapy in risk factor management: a mechanistic perspective. AB - Endothelial dysfunction contributes to mechanisms of atherogenesis and its clinical manifestations, including coronary heart disease. Cardiovascular risk factors have been linked directly to a loss of endothelial function, such as endothelium-dependent nitric oxide (NO) release, resulting in abnormal vasodilation in response to various stimuli. There is evidence that multiple risk factors, including hypertension and hyperlipidemia, lead to a synergistic effect on endothelial dysfunction, likely through oxidative stress mechanisms. Damage to the endothelium leads to reduced NO bioavailability and facilitates vessel wall permeability to low-density lipoprotein. Certain agents, including the antihypertensive drug amlodipine and the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin) atorvastatin, are known to influence endothelial function and NO bioavailability directly; these properties may contribute to clinical benefits. Recent experimental evidence at the cellular level indicates that these agents stimulate NO release from human endothelial cells in a highly synergistic fashion. The clinical implications of these observations are discussed in this article in the context of cardiovascular risk factor management strategies. PMID- 16356810 TI - Potential vascular benefits of statins. AB - Atherosclerosis is associated with a number of functional abnormalities that affect endothelium-dependent vasomotor function, inflammation, and thrombosis. The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) have effects on many of these functions, likely explaining their benefit in reducing the incidence of clinical events in patients at high risk of cardiovascular disease. Statins may improve this vascular biology by lowering levels of low-density lipoprotein (LDL) or potentially by a number of non-LDL related mechanisms. Cell culture and some animal studies have demonstrated LDL independent effects of statins. The non-LDL mechanisms include effects on isoprenoid production and function, interactions between caveolin and nitric oxide synthase, and direct immunomodulatory effects. Although these mechanisms are clearly demonstrated in the experimental setting, their relevance to the clinical use of statins is unknown. From a purely pragmatic viewpoint, the debate of lipid versus nonlipid effects of statins matters little to clinical practice. Their proven effect on vascular biology and risk reduction justifies their important therapeutic role. PMID- 16356811 TI - Diagnosis of suspected venous thromboembolic disease in pregnancy. AB - Venous thromboembolic disease is a leading cause of maternal mortality during pregnancy. Early and accurate radiological diagnosis is essential as anticoagulation is not without risk and clinical diagnosis is unreliable. Although the disorder is potentially treatable, unnecessary treatment should be avoided. Most of the diagnostic imaging techniques involve ionizing radiation which exposes both the mother and fetus to finite radiation risks. There is a relative lack of evidence in the literature to guide clinicians and radiologists on the most appropriate method of assessing this group of patients. This article will review the role of imaging of suspected venous thromboembolic disease in pregnant patients, highlight contentious issues such as radiation risk, intravenous contrast use in pregnancy and discuss the published guidelines, as well as suggesting an appropriate imaging algorithm based on the available evidence. PMID- 16356812 TI - Central venous catheters: the role of radiology. AB - The insertion and management of long-term venous catheters have long been the province of anaesthetists, intensive care physicians and surgeons. Radiologists are taking an increasing role in the insertion of central venous catheters (CVCs) because of their familiarity with the imaging equipment and their ability to manipulate catheters and guide-wires. The radiological management of the complications of CVCs has also expanded as a result. This article reviews the role of radiology in central venous access, covering the detection and management of their complications. PMID- 16356813 TI - Three-dimensional gadolinium-enhanced magnetic resonance breath-hold FLASH imaging in the diagnosis and staging of renal cell carcinoma. AB - Renal cell carcinoma accounts for 2% of all reported cancers. Its apparent incidence is increasing due to the more widespread use of cross-sectional imaging and as a result, tumours are being detected at an earlier stage. It is hoped that this improvement in early detection will result in a significant increase in survival rates. Radiological diagnosis and staging have a critical role in triaging patients' -treatment. Although computed tomography (CT) and ultrasound are well established in the evaluation of renal cell carcinoma, magnetic resonance (MR) techniques are still rapidly developing. In our institution breath hold three-dimensional (3D) gadolinium-enhanced fast low-angled single shot (FLASH) spoiled gradient-echo sequence imaging has become an integral part of staging for renal cell carcinoma. In this article, we review our experience of the use of this emerging technique in the diagnosis and staging of renal cancer. PMID- 16356814 TI - Multislice CT enteroclysis: technique and clinical applications. AB - Computed tomography (CT) enteroclysis was introduced to overcome the individual deficiencies of barium enteroclysis and abdominal CT. The use of multislice CT technology has made the procedure a feasible extension of the intubation infusion method of small bowel examination. This review is an update of the technique and clinical application of CT enteroclysis with illustrations. PMID- 16356815 TI - The radiological appearances of thalassaemia. AB - The skeletal changes of untreated thalassaemia result from ineffective erythropoiesis and expansion of the bone marrow and affect every part of the skeleton. These changes include osteoporosis, growth retardation, platyspondyly and kyphosis. Erythropoiesis occurs at extra-medullary sites, most commonly resulting in a paraspinal mass but occasionally affecting organs containing pluripotential stem cells. Repeated transfusion unaccompanied by iron chelation causes haemosiderosis; iron is deposited at various sites causing functional impairment. Iron-chelation therapy with desferrioxamine (DFX) prevents haemosiderosis but causes a skeletal dysplasia predominantly affecting the rapidly growing long bones, in particular the distal ulna, and causing irregularity and sclerosis of the physeal-metaphyseal junction and splaying of the metaphysis. DFX also exacerbates the observed growth retardation. DFX-induced skeletal dysplasia may herald toxicity, which is associated with visual and auditory impairment. Therefore, careful balancing of the transfusion regimen and iron-chelation therapy is required. Magnetic resonance imaging (MRI) is the most sensitive technique for the detection of DFX-induced dysplasia. PMID- 16356817 TI - Percutaneous abscess drainage in the UK: A national survey and single centre study. AB - AIM: To establish the current practice for management of radiologically placed percutaneous drains for abdominal sepsis in the UK and prospectively study the management of radiologically placed drains at our institution. METHOD: A questionnaire on the management of radiologically placed drains was sent to all radiology departments on a Royal College of Radiologists database. We prospectively followed all drains placed by our radiology department for drainage of abdominal collections, over a 7-month period. RESULTS: A total of 210 questionnaires were sent for the national survey, of these 117 were returned (55.7%). The majority of departments (70.5%) reported that after drain insertion the clinical team took over daily management. Just over 5% of departments either formally managed the drain or obtained final outcome data. From October 2003 to April 2004 we followed 63 consecutive drains placed in 45 patients, for abdominal sepsis. Thirty-nine drains (61.9%) were curative and 17 (26.9%) drains failed. Three drains (4.8%) were placed for palliation, and four drains (6.4%) were placed in order to temporise prior to surgery. Forty-three (68.3%) drains had a successful primary outcome: success after secondary percutaneous abscess drainage (PAD) improved to 46 (73.0%) drains. Two (3%) major complications occurred. CONCLUSIONS: The current approach in the UK to management of radiologically placed drains differs significantly from that practised in the USA. The most common type of support offered by radiology departments in the UK is of informal advice and follow-up, with the clinical team managing the patient's drain. Observations in our hospital highlighted problems relating to drain management that may impact on the success of PAD. We suggest that more formal radiological support after PAD would improve communication and potentially improve outcomes. PMID- 16356818 TI - Distance between the rectal wall and mesorectal fascia measured by MRI: Effect of rectal distension and implications for preoperative prediction of a tumour-free circumferential resection margin. AB - AIM: To determine the effect of rectal distension, used by some workers to facilitate staging, on mesorectal tissues. SUBJECTS AND METHODS: Ninety-seven consecutive rectal cancer staging MRI examinations were identified of which 76 were analysable: 48 studies were performed using rectal insufflation of 100 ml room air and 28 were performed without distension. Median age was 69 and 72 years, respectively. In each patient a single experienced observer measured the distance from the outer rectal wall to the inner margin of the mesorectal fascia at four locations (12, 3, 6 and 9 o'clock), excluding sites of tumour involvement, from the T1-weighted axial image at the level of the sacro-coccygeal junction. The two groups of measurements were compared using Mann-Whitney test statistic, and frequencies then categorized into <5 mm or > or =5 mm, and compared using Fisher's exact test. RESULTS: The median distance between the rectal wall and mesorectal fascia in the distended group was approximately half that found in the non-distended group, and significantly lower at the 3, 6 and 9 o'clock positions (p<0.001). 68/167 (41%) of measurements were 5mm or less, compared with 19/104 (18%) in the non-distended group (p<0.001). CONCLUSION: Rectal distension before MRI significantly reduces the distance between the rectal wall and mesorectal fascia. Although this is advocated to facilitate visualization of the primary tumour, it potentially affects the accuracy with which a clear circumferential resection margin can be predicted. PMID- 16356819 TI - Suspected and incidental pulmonary embolism on multidetector-row CT: analysis of technical and morphological factors influencing the diagnosis in a cross sectional cancer centre patient cohort. AB - AIM: To assess technical and computed tomography (CT) predictors of true-positive (TP) and false-negative (FN) radiological diagnoses in a retrospective patient cohort with pulmonary embolism (PE) in the setting of a cancer centre. MATERIALS AND METHODS: Two thousand, four hundred and twelve consecutive chest multidetector-row CT images from 1869 patients were reviewed for presence of PE. CT protocols and TP and FN radiological reports were determined and the clinical files reviewed for suspicion of PE. The severity of PE was assessed by an arterial obstruction index. Ancillary pulmonary findings were scored qualitatively and on a lobar basis. Statistical tests included analysis of variance and univariate and multivariate logistic regressions. RESULTS: Ninety one out of a total of 111 PE-positive images were included. Thirty-five patients had clinically suspected PE; 56 were not suspected of having PE. Forty-eight patients had TP diagnoses; 43 (39 of whom were not suspected of having PE) had FN radiological diagnoses. FN diagnoses were most frequent in oesophageal (17/19; 89.5%) and standard chest CT (19/35; 54.3%). Pulmonary CT angiography was associated with TP diagnosis (p<0.0001), whereas oesophageal CT was associated with FN diagnosis (p=0.001). Artefacts and arterial attenuation did not influence PE detection (p=0.017 and 0.066 for artefacts and arterial attenuation, respectively). However, the arterial obstruction index strongly predicted PE diagnosis (p=0.001). This was confirmed on multivariate analysis (p=0.041, 0.027 and 0.020 for pulmonary CT angiography, oesophageal protocols and arterial obstruction index, respectively). When stratified for clinically unsuspected cases, the arterial obstruction index remained the only predictor of PE diagnosis (p=0.009). CONCLUSION: Predictors of PE diagnosis were PE severity and technical factors; the latter were linked to clinical suspicion of PE. Arterial enhancement appears unlikely to contribute to missed diagnoses, if judged adequate for diagnosis of PE, and ancillary chest findings are unlikely to improve embolus detection. PMID- 16356820 TI - Clinically unsuspected pulmonary embolism--an important secondary finding in oncology CT. AB - AIM: To determine the rate of finding incidental pulmonary embolisms (PE) at staging or follow-up chest computed tomography (CT) in oncology patients. MATERIALS AND METHODS: Three hundred and eighty-five consecutive chest CT examinations, performed in patients with cancer, were prospectively assessed during a 9-month period between October 2003 and June 2004. These were spiral acquisitions between 5 and 8mm collimation acquired 25s after intravenous contrast medium administration. PE was diagnosed if a filling defect was seen in the central pulmonary arteries on two or more consecutive slices. RESULTS: Ten of 385 (2.6%) of these patients had incidentally detected PE. This was not attributable to any specific malignancy or chemotherapeutic regimen. These emboli were all central, between the main pulmonary artery and the lobar level. Although the significance of these findings in patients not clinically suspected of having PE could be contentious, all the patients were started on therapeutic anticoagulation as a result of this observation. CONCLUSION: Over one in 40 oncology patients have incidental central PE visible on the CT images performed to assess their malignancy. Formal review of the pulmonary arteries, using a work station, is advised in patients with malignancy. PMID- 16356821 TI - Diagnostic and interventional radiology workload in acute pancreatitis in an ITU/HDU setting. AB - AIM: To determine the impact on diagnostic and interventional radiology services when imaging patients with severe pancreatitis on intensive therapy (ITU) and high-dependency units (HDU) in a tertiary referral centre. MATERIAL AND METHODS: One hundred and sixty-nine patients admitted to ITU/HDU over a 9-year period (1996-2004) with severe acute pancreatitis were reviewed. There were 109 admissions to the ITU with length of stay of 0.2-81.6 days (mean 19.7 days) and 92 admissions to the HDU with length of stay of 0.4-12.8 days (mean 4.9 days). RESULTS: One hundred and seventy-nine computed tomography (CT) and 199 ultrasound (US) examinations were performed on the ITU patients in whom interventional procedures were required in 24% of patients undergoing CT examinations and in 32% of patients undergoing US. Sixty-two CT and 60 US examinations were performed in the HDU patients. The percentage of interventional procedures performed in HDU patients was similar to that in ITU patients, i.e., 18% CT-guided and 35% US guided. The proportion of patients that underwent investigations and interventions has gradually increased over the period of the study. Inpatient mortalities were 29% and 5.4%, respectively, in ITU and HDU patients. CONCLUSIONS: This study demonstrates the huge input and increasing workload undertaken by radiologists when managing patients with severe acute pancreatitis in an ITU/HDU setting. We believe this is partly due to the implementation of the British Society of Gastroenterology (BSG) guidelines on management of acute pancreatitis and partly due to the more intensive non-surgical management offered to patients being referred into a specialist tertiary referral unit. PMID- 16356822 TI - Trends in the on-call workload of radiologists. AB - AIM: To evaluate the radiologist on-call clinical workload of an inner-city acute teaching hospital. MATERIALS AND METHODS: Data were collected prospectively from a 9-week assessment of the workload of the radiologists on-call at our Trust. Data collection was undertaken using a questionnaire-based survey detailing the date and time of request, and where appropriate, the imaging examination performed. The referring clinician's grade and speciality were also recorded. The results obtained were compared with a similar study carried out in 1996. RESULTS: A total of 160 requests for out-of-hours imaging were received over the 9-week period. CONCLUSIONS: The figures indicate that on-call work is increasing with an approximate doubling of out-of-hours requests compared with the survey performed 7 years earlier. PMID- 16356823 TI - Physical and cognitive task analysis in interventional radiology. AB - AIM: To identify, describe and detail the cognitive thought processes, decision making, and physical actions involved in the preparation and successful performance of core interventional radiology procedures. MATERIALS AND METHODS: Five commonly performed core interventional radiology procedures were selected for cognitive task analysis. Several examples of each procedure being performed by consultant interventional radiologists were videoed. The videos of those procedures, and the steps required for successful outcome, were analysed by a psychologist and an interventional radiologist. Once a skeleton algorithm of the procedures was defined, further refinement was achieved using individual interview techniques with consultant interventional radiologists. Additionally a critique of each iteration of the established algorithm was sought from non participating independent consultant interventional radiologists. RESULTS: Detailed task descriptions and decision protocols were developed for five interventional radiology procedures (arterial puncture, nephrostomy, venous access, biopsy-using both ultrasound and computed tomography, and percutaneous transhepatic cholangiogram). Identical tasks performed within these procedures were identified and standardized within the protocols. CONCLUSIONS: Complex procedures were broken down and their constituent processes identified. This might be suitable for use as a training protocol to provide a universally acceptable safe practice at the most fundamental level. It is envisaged that data collected in this way can be used as an educational resource for trainees and could provide the basis for a training curriculum in interventional radiology. It will direct trainees towards safe practice of the highest standard. It will also provide performance objectives of a simulator model. PMID- 16356824 TI - Pseudoaneurysm after aortic coarctation repair presenting with an aortobronchial fistula successfully treated with an aortic stent graft. PMID- 16356825 TI - The neuropilins and their role in tumorigenesis and tumor progression. AB - The neuropilins were originally described as receptors for the six axon guidance factors belonging to the class-3 semaphorins. They were subsequently found to function in addition as receptors for specific splice forms of angiogenic factors belonging to the VEGF family. The neuropilins are expressed in many types of cancer cells, in endothelial cells and in additional many types of normal diploid cell types. Recent findings indicate that the neuropilins and their associated plexin and tyrosine-kinase VEGF receptors play a regulatory role in developmental angiogenesis as well as in tumor angiogenesis. The neuropilin ligands belonging to the semaphorin family as well as the various VEGF's function as modulators of angiogenesis and tumor angiogenesis. Furthermore, since many types of cancer cells express neuropilins and neuropilin associated receptors, it is not surprising that various neuropilin ligands can modulate the behavior of cancer cells directly leading to the potentiation or inhibition of tumor progression. PMID- 16356826 TI - Molecular chaperones throughout the life cycle of the androgen receptor. AB - Aberrant signaling by the androgen receptor contributes to the initiation and progression of prostate cancer. The involvement of molecular chaperones in the processes of folding, activation, trafficking, and transcriptional activity of the androgen receptor provide different points along the signaling axis where regulation of androgen receptor activity can be hijacked to provide growth signals for clonal selection in cancer progression. Evidence exists of abnormal chaperone expression that could contribute to the upregulation of AR activity in prostate tumors. Regardless of whether chaperones are involved in the causation of prostate carcinogenesis, molecular chaperones provide therapeutic targets for the treatment of prostate cancer. PMID- 16356827 TI - Mycophenolic acid-induced replication arrest, differentiation markers and cell death of androgen-independent prostate cancer cells DU145. AB - Inosine 5'-monophosphate dehydrogenase inhibitors including mycophenolic acid (MPA) are effective inducers of terminal differentiation in a variety of distinct human tumor cell types. Here, we report that MPA also induces such a differentiation in the androgen-independent prostate cancer derived cell line DU145. MPA evoked replication arrest and accumulation of the DU145 cells in the S phase of the cell cycle. The inhibitor also induced the expression of CD55, clusterin, granulophysin, glucose-regulated protein 78, vasoactive intestinal polypeptide and prostate-specific transglutaminase, which are differentiation markers associated with the phenotype of normal prostate cells. We suggest that inosine 5'-monophosphate dehydrogenase inhibitors, which are already used for the treatment of other diseases, may be used as potential differentiation therapy drugs to control prostate cancer. PMID- 16356828 TI - Modulation of arginine metabolic pathways as the potential anti-tumor mechanism of recombinant arginine deiminase. AB - Arginine deiminase (ADI), currently in clinical trials, has various biological activities including anti-proliferation, anti-angiogenesis and inhibition of inducible nitric oxide synthase (iNOS). To recognize limitations and therapeutic applications, the mechanism of ADI modulation of arginine metabolic pathways was investigated. MCF-7 and A549 cells have notable different sensitivity to recombinant ADI (rADI) and express diverse argininosuccinate synthase (AS) activity, which regenerates arginine. Due to compartmentalization of arginine, utilization of arginine for protein synthesis occurs from either the intracellular arginine pool or the citrulline-arginine-regeneration pathway, whereas for polyamine synthesis, utilization is only from the intracellular arginine pool. Modulating AS activity or introducing rADI intracellularly to reduce intracellular arginine regeneration may expand therapeutic applications of rADI. PMID- 16356829 TI - Myricetin stimulates the absorption of the pro-carcinogen PhIP. AB - The effect of the flavonoid myricetin on the transport of the pro-carcinogen 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) through differentiated Caco 2 monolayers, a model for the intestinal epithelium, is described. Myricetin causes an increase of the transport of PhIP from the apical to the basolateral compartment. This effect was observed at physiologically relevant concentrations of PhIP and myricetin. Cyclosporin A (MRP2 inhibitor) but not PSC833 (P-gp inhibitor) showed a similar effect on PhIP transport. The results indicate that myricetin induces an increased basolateral uptake of the pro-carcinogen PhIP, in part through inhibition of the MRP2 mediated excretion of PhIP from the intestinal cells back to the lumen. PMID- 16356830 TI - Oral artemisinin prevents and delays the development of 7,12 dimethylbenz[a]anthracene (DMBA)-induced breast cancer in the rat. AB - Artemisinin, a compound isolated from the sweet wormwood Artemisia annua L., has previously been shown to have selective toxicity towards cancer cells in vitro. In the present experiment, we studied the potential of artemisinin to prevent breast cancer development in rats treated with a single oral dose (50mg/kg) of 7,12-dimethylbenz[a]anthracene (DMBA), known to induce multiple breast tumors. Starting from the day immediately after DMBA treatment, one group of rats was provided with a powdered rat-chow containing 0.02% artemisinin, whereas a control group was provided with plain powdered food. For 40 weeks, both groups of rats were monitored for breast tumors. Oral artemisinin significantly delayed (P<.002) and in some animals prevented (57% of artemisinin-fed versus 96% of the controls developed tumors, P<.01) breast cancer development in the monitoring period. In addition, breast tumors in artemisinin-fed rats were significantly fewer (P<.002) and smaller in size (P<.05) when compared with controls. Since artemisinin is a relatively safe compound that causes no known side effects even at high oral doses, the present data indicate that artemisinin may be a potent cancer chemoprevention agent. PMID- 16356831 TI - 2-Methoxyestradiol and paclitaxel have similar effects on the cell cycle and induction of apoptosis in prostate cancer cells. AB - 2-Methoxyestradiol (2-ME) is an endogenous metabolite of estradiol with promise for cancer chemotherapy, including advanced prostate cancer. We have focused on events related to cell cycle arrest (G1 and G2/M) and induction of apoptosis in human prostate cancer cells. Treatment with 2-ME increased cyclin B1 protein and its associated kinase activity followed by later inhibition of cyclin A-dependent kinase activity and induction of apoptosis. Similar results were obtained with paclitaxel (taxol), a clinically relevant agent used to treat advanced prostate cancer. Cyclin-dependent kinase inhibitors prevented 2-ME and paclitaxel-mediated increase in cyclin B1-dependent kinase activity and blocked induction of apoptosis. Reduction of X-linked inhibitor of apoptosis (XIAP) protein by 2-ME and paclitaxel correlated with increased apoptosis. Lower doses of 2-ME and paclitaxel resulted in G1 (but not G2/M) cell cycle arrest in the p53 wild type LNCaP cell line, but with minimal induction of apoptosis. We suggest that 2-ME and paclitaxel-mediated induction of apoptosis in prostate cancer cells requires activation of cyclin B1-dependent kinase that arrests cells in G2/M and subsequently leads to the induction of apoptotic cell death. PMID- 16356832 TI - Establishment and characterization of murine small cell lung carcinoma cell lines derived from HPV-16 E6/E7 transgenic mice. AB - We have established two murine cell lines derived from Small Cell Lung Carcinomas (SCLCs) developed by HPV-E6/E7 transgenic mice. These cells named PPAP-9 and PPAP 10 were isolated from mice bearing tumors, 9 and 10 months old, respectively. The cells, 5 microm in diameter, express HPV oncoproteins and sustain tumor formation after subcutaneous injection in syngenic mice. A detailed analysis indicated the epithelial origin and the neuroendocrine differentiation of these cells. We showed by confocal immunofluorescence the expression of the epithelial marker cytokeratin 5, whose gene promoter was used to direct the expression of HPV E6/E. Cells express several neuroendocrine markers such as CGRP, MAP-2, Ash1, CgrA, Scg2. The neuroendocrine differentiation of these cells was further confirmed by electron microscopy demonstrating neuropeptides secreting granules in their cytoplasm. Furthermore, in agreement with the altered expression observed in the majority of human SCLC we showed in these cells the absence of both p53 and pRB and a dramatic reduction in the expression of Caveolin-1. PMID- 16356833 TI - A cDNA-microarray analysis of camptothecin resistance in glioblastoma cell lines. AB - Chemotherapy, as generally available, is of a limited value in curing malignant brain tumors (gliomas), which often develop resistance to drugs, becoming completely unresponsive to any standard therapeutic approach. Camptothecins, a family of topoisomerase I inhibitor drugs, represent a new promising treatment strategy and are currently under evaluation for testing the clinical efficacy. We selected a CPT-resistant sub-line (U87CPT-R) from U87-MG grade III-IV astrocytoma cells, and compared the expression profile of the two cell lines by cDNA microarray, as a preliminary screening of the molecular mechanisms involved in the acquisition of CPT resistance in glioma cells. The relevant role of IL-1 beta overproduction as well as a generalised up-regulation of genes implicated in angiogenesis and inflammatory response are discussed in details. PMID- 16356834 TI - Role of DNA hypomethylation in the development of the resistance to doxorubicin in human MCF-7 breast adenocarcinoma cells. AB - The resistance of cancer cells to chemotherapeutic agents is a major clinical problem and an important cause of treatment failure in cancer. Mechanisms that have developed to guard cancer cells against anti-cancer drugs are major barriers to successful anti-cancer therapy. Therefore, the identification of novel mechanisms of cellular resistance holds the promise of leading to better treatments for cancer patients. In the present study, we used human MCF-7 breast adenocarcinoma cell line and its doxorubicin-resistant variant MCF-7/R to determine the role of alterations of DNA methylation of chemoresitance-related genes, such as multidrug resistance 1 (MDR1), glutathione-S-transferase (GSTpi), O(6)-methylguanine DNA methyltransferase (MGMT), and urokinase (Upa), in the development of drug resistance. The promoter regions of MDR1, GSTpi, MGMT, and Upa genes were highly methylated in MCF-7 cell line but not in its MCF-7/R drug resistant variant. The hypomethylated status of MDR1 gene was associated with overexpression of P-glycoprotein. We hypothesize that acquirement of doxorubicin resistance of MCF-7 cells is associated with DNA hypomethylation of the promoter regions of the MDR1, GSTpi, MGMT, and Upa genes. PMID- 16356835 TI - Tp53-gene transfer induces hypersensitivity to low doses of X-rays in glioblastoma cells: a strategy to convert a radio-resistant phenotype into a radiosensitive one. AB - Tp53 is frequently mutated or inactivated in glioblastomas. Due to the impairment of p53 activity, glioblastomas show a high degree of radioresistance. In an attempt to convert the radioresistant phenotype to a more radiosensitive one, we evaluated the efficacy of the combination of Adp53 gene transfer and X-ray irradiation. The combination of Adp53, at low multiplicity in order to mimic the low in vivo efficiency of virus-mediated gene delivery, with X-ray irradiation resulted in a marked decrease of glioblastoms cell survival. Interestingly, Adp53 was able to induce low dose (<2Gy) hypersensitivity. The data suggest the possibility for the development of new therapeutic strategies. PMID- 16356836 TI - Resveratrol induces apoptosis and inhibits angiogenesis in human breast cancer xenografts in vivo. AB - Resveratrol, a polyphenol found in grapes and wine, is considered a potential cancer chemopreventive agent. Resveratrol has been shown to induce transcription via both ERalpha and ERbeta. We observed significantly lower tumor growth, decreased angiogenesis, and increased apoptotic index in ERalpha- ERbeta+ MDA-MB 231 tumors in resveratrol-treated nude mice compared with controls. In vitro we found a significant increase in apoptosis in resveratrol-treated MDA-MB-231 cells in addition to significantly reduced extracellular levels of VEGF. This study supports the potential use of resveratrol as a chemotherapeutic agent in breast cancers. PMID- 16356837 TI - Administration of human umbilical cord blood cells delays the onset of prostate cancer and increases the lifespan of the TRAMP mouse. AB - Stem cell transplantation to improve the onset and survival of animals or humans with prostate cancer has not been studied adequately. In this study, we examined whether intravenous administration of human umbilical cord blood (HUCB) mononuclear cells into TRAMP (transgenic adenocarcinoma of the mouse prostate) mice can delay the onset of prostate cancer and improve survival of these mice before and after the development of cancer. Twenty TRAMP mice were randomly divided into 2 groups. One group of 10 mice received 200 x10(6) HUCB mononuclear cells retro-orbitally into the venous plexus at the age of 6 weeks. Another group of 10 mice did not receive HUCB cells and served as control mice. The presence of tumor was detected by abdominal palpation, which was confirmed by biopsy. When 4 of the 10 control mice developed the tumor, they were treated with the same dose of HUCB cells. Either at the time of death or sacrifice, various tissues were examined for the presence of HUCB cell total RNA by reverse transcriptase PCR. Also, the tissues were examined histologically for the presence of metastasis and carcinoma. Kaplan-Meier survival plots were used to assess the lifespan of the mice. The data show that the control mice developed the tumor much earlier than the treated mice (control vs treated: 238+/-38 vs 311+/-40 days; P<0.001). Also, transplantation of HUCB cells either before or after the development of tumor significantly increased the life span compared to that of control mice. Persistence of human RNA either in blood or spleen was associated with prolonged survival. No graft vs host disease was observed in any of the mice. In conclusion, transplantation of HUCB mononuclear cells via intravenous administration into TRAMP mice retards not only the development of prostate cancer but also increases the lifespan of these mice. PMID- 16356838 TI - Expression of sFRP-4 and beta-catenin in human colorectal carcinoma. AB - Alterations to the Wnt signalling pathway occur in the majority of colorectal cancers and result in abnormal accumulation of beta-catenin. The secreted frizzled related proteins (sFRPs) are antagonists that bind Wnt and inhibit signalling along this pathway. We investigated expression of the sFRP family member, sFRP-4, and beta-catenin in 1,044 human colorectal carcinomas using tissue microarrays and immunohistochemistry. Both proteins showed markedly increased expression levels in tumors compared to normal mucosa, but no significant associations with pathological features or with patient outcome. sFRP 4 was co-expressed with beta-catenin, p53, and COX-2, while the absence of beta catenin expression was strongly associated with loss of expression of the MLH1 mismatch repair gene. In contrast to other sFRP family members, sFRP-4 expression appears to be upregulated in colorectal carcinoma. PMID- 16356839 TI - Formation of DHP-derived DNA adducts from metabolic activation of the prototype heliotridine-type pyrrolizidine alkaloid, lasiocarpine. AB - Pyrrolizidine alkaloids (PAs) are probably the most common poisonous plants affecting livestock, wildlife, and humans. The PAs that have been found to be tumorigenic in experimental animals belong to the retronecine-, heliotridine-, and otonecine-type PAs. Our recent mechanistic studies indicated that riddelliine, a tumorigenic retronecine-type PA, induced tumors via a genotoxic mechanism mediated by the formation of 6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H pyrrolizine (DHP)-derived DNA adducts. The same adducts were formed from clivorine, a tumorigenic otonecine-type PA from metabolism of clivorine by rat liver microsomes in the presence of calf thymus DNA. In this study, we report that metabolism of lasiocarpine, the prototype heliotridine PA, by F344 rat liver microsomes resulted in the formation of DHP. When incubated in the presence of calf thymus DNA, the same DHP-derived DNA adducts were formed. These results suggest that these DHP-derived DNA adducts are potential biomarkers of exposure and tumorigenicity for all types of PAs. PMID- 16356840 TI - Biotechnology may help thwart malaria. PMID- 16356841 TI - The laboratory in a droplet. AB - In this issue of Chemistry and Biology, the groups of Tawfik and Griffiths present fluorescence-activated cell sorting of double emulsions as a generally applicable screen for enzyme activity. This novel methodology increases the throughput of a typical enzyme screen by two orders of magnitude. PMID- 16356842 TI - Acyclic peptide inhibitors of amylases. AB - In this issue of Chemistry and Biology, a library screening approach reveals a linear octapeptide inhibitor of alpha-amylases reached by de novo design . The selected molecule shares characteristics with naturally occurring protein inhibitors -- a result that suggests general rules for the design of peptide based amylase inhibitors may be achievable. PMID- 16356843 TI - Studies on calcium dependence reveal multiple modes of action for triptolide. AB - Triptolide, a diterpene triepoxide isolated from the traditional Chinese medicinal vine Trypterygium wilfordii hook f., has been shown to induce rapid apoptosis in a myriad of cancer cell lines and inhibit NFkappaB transactivation. To understand further the general cellular mechanisms for this therapeutically relevant natural product, binding and biological activities were assessed. Studies showed that triptolide binding was saturable, reversible, and primarily localized to cell membranes. Depletion of calcium enhanced overall binding while differentially modulating biological function. Furthermore, triptolide's structural moieties demonstrated variability in the regulation of cell death versus inhibition of NFkappaB transactivation. These results implicate triptolide in the manipulation of at least two distinct cellular pathways with differing requirements for calcium and effective triptolide concentration in order to elicit each particular biological function. PMID- 16356844 TI - Microtubule interactions with chemically diverse stabilizing agents: thermodynamics of binding to the paclitaxel site predicts cytotoxicity. AB - The interactions of microtubules with most compounds described as stabilizing agents have been studied. Several of them (lonafarnib, dicumarol, lutein, and jatrophane polyesters) did not show any stabilizing effect on microtubules. Taccalonolides A and E show paclitaxel-like effects in cells, but they were not able to modulate in vitro tubulin assembly or to bind microtubules, which suggests that other factors are involved in their cellular effects. The binding constants of epothilones, eleutherobin, discodermolide, sarcodictyins, 3,17beta diacetoxy-2-ethoxy-6-oxo-B-homo-estra-1,3,5(10)-triene, and dictyostatin to the paclitaxel site; the critical concentrations of ligand-induced assembly; and their cytotoxicity in carcinoma cells have been measured, and correlations between these parameters have been determined. The inhibition of cell proliferation correlates better with the binding enthalpy change than with the binding constants, suggesting that large, favorable enthalpic contribution to the binding is desired to design paclitaxel site drugs with higher cytotoxicity. PMID- 16356845 TI - High-throughput screening of enzyme libraries: thiolactonases evolved by fluorescence-activated sorting of single cells in emulsion compartments. AB - Single bacterial cells, each expressing a different library variant, were compartmentalized in aqueous droplets of water-in-oil (w/o) emulsions, thus maintaining a linkage between a plasmid-borne gene, the encoded enzyme variant, and the fluorescent product this enzyme may generate. Conversion into a double, water-in-oil-in-water (w/o/w) emulsion enabled the sorting of these compartments by FACS, as well as the isolation of living bacteria cells and their enzyme coding genes. We demonstrate the directed evolution of new enzyme variants by screening >10(7) serum paraoxonase (PON1) mutants, to yield 100-fold improvements in thiolactonase activity. In vitro compartmentalization (IVC) of single cells, each carrying >10(4) enzyme molecules, in a volume of <10 femtoliter (fl), enabled detection and selection despite the fast, spontaneous hydrolysis of the substrate, the very low initial thiolactonase activity of PON1, and the use of difusable fluorescent products. PMID- 16356846 TI - High-throughput screening of enzyme libraries: in vitro evolution of a beta galactosidase by fluorescence-activated sorting of double emulsions. AB - We describe a completely in vitro high-throughput screening system for directed evolution of enzymes based on in vitro compartmentalization (IVC). Single genes are transcribed and translated inside the aqueous droplets of a water-in-oil emulsion. Enzyme activity generates a fluorescent product and, after conversion into a water-in-oil-in-water double emulsion, fluorescent droplets are sorted using a fluorescence-activated cell sorter (FACS). Earlier in vivo studies have demonstrated that Ebg, a protein of unknown function, can evolve to allow Escherichia coli lacking the lacZ beta-galactosidase gene to grow on lactose. Here we demonstrate that we can evolve Ebg into an enzyme with significant beta galactosidase activity in vitro. Only two specific mutations were ever seen to provide this improvement in Ebg beta-galactosidase activity in vivo. In contrast, nearly all the improved beta-galactosidases selected in vitro resulted from different mutations. PMID- 16356847 TI - An iterative type I polyketide synthase PKSN catalyzes synthesis of the decaketide alternapyrone with regio-specific octa-methylation. AB - A biosynthetic gene cluster containing five genes, alt1-5, was cloned from Alternaria solani, a causal fungus of early blight disease to tomato and potato. Homology searching indicated that the alt1, 2, and 3 genes code for cytochrome P450s and the alt4 gene for a FAD-dependent oxygenase/oxidase. The alt5 gene encodes a polyketide synthase (PKS), named PKSN, that was found to be an iterative type I complex reduced-type PKS with a C-methyltransferase domain. To identify the PKSN function, the alt5 gene was introduced into the fungal host Aspergillus oryzae under an alpha-amylase promoter. The transformant produced a polyketide compound, named alternapyrone, whose structure is shown to be 3,5 dimethyl-4-hydroxy-6-(1,3,5,7,11,13-hexamethyl-3,5,11-pentadecatrienyl)-pyran-2 one. Labeling experiments confirmed that alternapyrone is a decaketide with octa methylation from methionine on every C(2) unit except the third unit. PMID- 16356848 TI - Chemoenzymatic synthesis of glycopeptides with PglB, a bacterial oligosaccharyl transferase from Campylobacter jejuni. AB - The gram-negative bacterium Campylobacter jejuni has a general N-linked glycosylation pathway encoded by the pgl gene cluster. One of the proteins in this cluster, PgIB, is thought to be the oligosaccharyl transferase due to its significant homology to Stt3p, a subunit of the yeast oligosaccharyl transferase complex. PgIB has been shown to be involved in catalyzing the transfer of an undecaprenyl-linked heptasaccharide to the asparagine side chain of proteins at the Asn-X-Ser/Thr motif. Using a synthetic disaccharide glycan donor (GaINAc alpha1,3-bacillosamine-pyrophosphate-undecaprenyl) and a peptide acceptor substrate (KDFNVSKA), we can observe the oligosaccharyl transferase activity of PgIB in vitro. Furthermore, the preparation of additional undecaprenyl-linked glycan variants reveals the ability of PgIB to transfer a wide variety of saccharides. With the demonstration of PgIB activity in vitro, fundamental questions surrounding the mechanism of N-linked glycosylation can now be addressed. PMID- 16356849 TI - The beta-carboline analog Mana-Hox causes mitotic aberration by interacting with DNA. AB - Mana-Hox, an analog of beta-carbolines with anticancer activity, induces aberrant mitosis and delays mitotic exit. However, the cellular target is not known. In this study, we visualized the intracellular localization of Mana-Hox. Mana-Hox rapidly penetrated into cells (within 1 min) and concentrated on disorganized metaphase chromosomes after 13 hr of exposure. We demonstrated that Mana-Hox is a noncovalent DNA binder that can interact with DNA through intercalation and/or through minor groove binding. Furthermore, Mana-Hox also inhibits topoisomerase II relaxation activity in vitro, suggesting that Mana-Hox could perturb mitotic chromosome decatenation. Overall, Mana-Hox binding to DNA plays a critical role in the induction of aberrant mitosis and contributes to its anticancer activity. PMID- 16356850 TI - Thiamine pyrophosphate riboswitches are targets for the antimicrobial compound pyrithiamine. AB - Thiamine metabolism genes are regulated in numerous bacteria by a riboswitch class that binds the coenzyme thiamine pyrophosphate (TPP). We demonstrate that the antimicrobial action of the thiamine analog pyrithiamine (PT) is mediated by interaction with TPP riboswitches in bacteria and fungi. For example, pyrithiamine pyrophosphate (PTPP) binds the TPP riboswitch controlling the tenA operon in Bacillus subtilis. Expression of a TPP riboswitch-regulated reporter gene is reduced in transgenic B. subtilis or Escherichia coli when grown in the presence of thiamine or PT, while mutant riboswitches in these organisms are unresponsive to these ligands. Bacteria selected for PT resistance bear specific mutations that disrupt ligand binding to TPP riboswitches and derepress certain TPP metabolic genes. Our findings demonstrate that riboswitches can serve as antimicrobial drug targets and expand our understanding of thiamine metabolism in bacteria. PMID- 16356851 TI - Chemistry and biology of moverastins, inhibitors of cancer cell migration, produced by Aspergillus. AB - Cancer cell migration is a required step in cancer metastasis. We screened for inhibitors of cancer cell migration of microbial origin, and obtained moverastin, a member of the cylindrol family, from Aspergillus sp. F7720. However, the results of an NMR spectroscopic analysis raised the possibility that moverastin is a mixture of two diastereomers. Separation of the C-10 epimers of synthetic moverastin and a bioassay revealed that both diastereomers (moverastins A and B) had inhibitory effects on cell migration. Furthermore, we demonstrated that moverastins A and B inhibited FTase in vitro, and they also inhibited both the membrane localization of H-Ras and the activation of the PI3K/Akt pathway in EC17 cells. Thus, moverastins inhibited the migration of tumor cells by inhibiting the farnesylation of H-Ras, and subsequent H-Ras-dependent activation of the PI3K/Akt pathway. PMID- 16356852 TI - De novo design of alpha-amylase inhibitor: a small linear mimetic of macromolecular proteinaceous ligands. AB - We report a low molecular weight inhibitor of alpha-amylases based on a linear peptidic scaffold designed de novo through the use of combinatorial chemistry. The inhibitory motif denoted PAMI (peptide amylase inhibitor) was selected by using L-peptide libraries and was fine-tuned by the introduction of unnatural modifications. PAMI specifically inhibits glycoside hydrolases of family 13. Its interaction with porcine pancreatic alpha-amylase was characterized by inhibition kinetics, fluorescence competition assays with natural alpha-amylase inhibitors, and isothermal titration calorimetry. We demonstrate that the critical amino acid residues in PAMI are shared with those in the macromolecular proteinaceous inhibitors that, however, bind to alpha-amylases through a spatially scattered set of intermolecular contacts. Thus, natural molecular evolution as well as combinatorial evolution selected the same alpha-amylase binding determinants for completely different spatial frameworks. PMID- 16356853 TI - Phosphorylation of CARMA1: the link(er) to NF-kappaB activation. AB - Antigen receptor-induced NF-kappaB activation depends on receptor-proximal and distal signaling events. Two papers in this issue of Immunity demonstrate that PKC-dependent phosphorylation of CARMA1 is the critical molecular link that controls the activation of the IKK signalosome and NF-kappaB. PMID- 16356854 TI - Poxviruses aren't stuPYD. AB - Pathogens utilize many strategies to dampen the host inflammatory response. In this issue of Immunity, a report by Johnston and colleagues reveals a poxvirus strategy that inhibits the inflammasome, arresting secretion of interleukin-1 related cytokines, thus silencing key alarms that mobilize host defenses. PMID- 16356855 TI - Phosphorylation of the CARMA1 linker controls NF-kappaB activation. AB - PKC isoforms and CARMA1 play crucial roles in immunoreceptor-dependent NF-kappaB activation. We tested whether PKC-dependent phosphorylation of CARMA1 directly regulates this signaling cascade. B cell antigen receptor (BCR) engagement led to the progressive recruitment of CARMA1 into lipid rafts and to the association of CARMA1 with, and phosphorylation by, PKCbeta. Furthermore, PKCbeta interacted with the serine-rich CARMA1 linker, and both PKCbeta and PKCtheta phosphorylated identical serine residues (S564, S649, and S657) within this linker. Mutation of two of these sites ablated the functional activity of CARMA1. In contrast, deletion of the linker resulted in constitutive, receptor- and PKC-independent NF kappaB activation. Together, our data support a model whereby CARMA1 phosphorylation controls NF-kappaB activation by triggering a shift from an inactive to an active CARMA1 conformer. This PKC-dependent switch regulates accessibility of the CARD and CC domains and controls assembly and full activation of the membrane-associated IkappaB kinase (IKK) signalosome. PMID- 16356856 TI - Phosphorylation of CARMA1 plays a critical role in T Cell receptor-mediated NF kappaB activation. AB - CARMA1 mediates T cell receptor (TCR)-induced NF-kappaB activation. However, how TCR links to CARMA1 in the signaling pathway is not clear. Here, we show that CARMA1 is inducibly phosphorylated after TCR-CD28 costimulation. This phosphorylation is likely induced by PKCtheta, since PKCtheta induces phosphorylation of CARMA1 in vitro and in vivo. Our results indicate that the PKCtheta-induced phosphorylation of CARMA1 likely occurs on Ser552 on the Linker region of CARMA1. Importantly, expression of CARMA1 mutant, in which Ser552 is mutated, fails to mediate TCR-induced NF-kappaB activation in CARMA1-deficient T cells. The functional defect of this CARMA1 mutant is likely due to the fact that this mutant cannot be phosphorylated at the critical residue, thereby failing to recruit the downstream signaling components into the immunological synapse. Together, our studies provide the first genetic evidence that the phosphorylation of CARMA1 plays a critical role in the TCR signaling pathway. PMID- 16356857 TI - A poxvirus-encoded pyrin domain protein interacts with ASC-1 to inhibit host inflammatory and apoptotic responses to infection. AB - Proinflammatory caspases play an essential role in innate immune responses to infection by regulating the cleavage and activation of proinflammatory cytokines. Activation of these enzymes requires the assembly of an intracellular molecular platform, termed the inflammasome, which is comprised of members of the pyrin domain (PYD)-containing superfamily of apoptosis and inflammation-regulatory proteins. We report here the identification and characterization of a poxvirus encoded PYD-containing protein that interacts with the ASC-1 component of the inflammasome and inhibits caspase-1 activation and the processing of IL-1beta and IL-18 induced by diverse stimuli. Knockout viruses that do not express this protein are unable to productively infect monocytes and lymphocytes due to an abortive phenotype and are markedly attenuated in susceptible hosts due to decreased virus dissemination and enhanced inflammatory responses at sites of infection. Thus, modulation of inflammasome function constitutes an important immunomodulatory strategy employed by poxviruses to circumvent host antiviral responses. PMID- 16356859 TI - Epidermal langerhans cell-deficient mice develop enhanced contact hypersensitivity. AB - Epidermal Langerhans cells (LCs), a distinct skin-resident dendritic cell population, acquire antigen in the skin and migrate to draining lymph nodes where they are thought to initiate adaptive immune responses. To examine the functional requirement of LCs in skin immunity, we generated BAC transgenic mice in which the regulatory elements from human Langerin were used to drive expression of diphtheria toxin. The resulting mice have a constitutive and durable absence of epidermal LCs but are otherwise intact. Unexpectedly, we found that contact hypersensitivity (CHS) was amplified rather than abrogated in the absence of LCs. Moreover, we showed that LCs act during the priming and not the effector phase. Thus, LCs not only were dispensable for CHS, but they served to regulate the response, a previously unappreciated function. PMID- 16356860 TI - Nef-mediated lipid raft exclusion of UbcH7 inhibits Cbl activity in T cells to positively regulate signaling. AB - Lentiviral Nef increases T cell signaling activity, but the molecular nature of the stimulus involved is incompletely described. We explored CD4 T cell lipid raft composition in the presence and absence of Nef. Here, the E2 ubiquitin conjugating enzyme UbcH7, which acts in conjunction with c-Cbl, is absent from lipid rafts. This Nef-mediated exclusion is associated with failure of ubiquitination of activated Vav. In the presence of Nef, lipid raft Cdc42 is activated and forms a ternary complex between the c-Cbl-interacting protein p85Cool-1/betaPix and c-Cbl, displacing UbcH7 from rafts. Suppression of p85Cool 1/betaPix expression restores UbcH7 raft localization and Vav ubiquitination and diminishes Cdc42 activity. Moreover, p85Cool-1/betaPix knockdown attenuates HIV replication. Thresholds for activation of signaling involve the intricate balance of positive and negative regulators. Here we provide evidence for Nef disruption of a negative regulator of T cell signaling in promoting HIV replication. PMID- 16356858 TI - Constitutively active beta-catenin confers multilineage differentiation potential on lymphoid and myeloid progenitors. AB - Beta-catenin-mediated Wnt signaling may contribute to the self-renewal of hematopoietic stem cells and proliferation in some malignancies. We now show that expression of constitutively active beta-catenin in normal lymphoid or myeloid progenitors generated uncommitted cells with multilineage differentiation potential. Inappropriate gene expression occurred in cells destined to produce either cell type and caused corresponding changes in their characteristics. For example, forced activation of beta-catenin quickly increased C/EBPalpha while reducing EBF and Pax-5 in lymphoid progenitors that then generated myeloid cells. Inversely, EBF dramatically increased in transduced myeloid progenitors and lymphocytes were produced. The results indicate that ectopic activation of beta catenin destabilizes lineage fate decisions and confers some, but not all, stem cell properties on committed progenitors. PMID- 16356861 TI - The juxtamembrane wedge negatively regulates CD45 function in B cells. AB - CD45 is a receptor-like protein tyrosine phosphatase highly expressed on all nucleated hematopoietic cells. We previously generated mice containing a point mutation in the juxtamembrane wedge of CD45. Demonstrating the critical negative regulatory function of the wedge, the CD45 E613R mutation led to a lymphoproliferative disorder (LPD) and a lupus-like autoimmune syndrome. Here we show the central role of B cells in this phenotype. Genetic elimination of B cells, but not T cells, ablates the LPD. In contrast to CD45-deficient B cells, the E613R mutation generates hyperresponsive B cells. Comparison of CD45 deficient and CD45 E613R mice reveals dichotomous effects of these mutations on B cell development. Together, the results support a role for CD45 as a rheostat, with both positive and negative regulatory functions, that fine-tunes the signal transduction threshold at multiple checkpoints in B cell development. PMID- 16356862 TI - Lymph node dendritic cells control CD8+ T cell responses through regulated FasL expression. AB - The lethal outcome of high-dose pulmonary virus infection is thought to reflect high-level, sustained virus replication and associated lung inflammation prior to development of an adaptive immune response. Herein, we demonstrate that the outcome of lethal/sublethal influenza infection instead correlates with the initial virus replication tempo. Furthermore, the magnitude of early lung antiviral CD8+ T cell responses varies inversely with inoculum dose and is controlled by lymph-node-resident dendritic cells (LNDC) through IL-12p40 regulated FasL-dependent T cell apoptosis. These results suggest that the inoculum dose and replication rate of a pathogen entering the respiratory tract may regulate the strength of the adaptive immune response, and the subsequent outcome of infection and that LNDC may serve as regulators (gatekeepers) in the development of CD8+ T cell responses. PMID- 16356863 TI - Structural and mutational analyses of a CD8alphabeta heterodimer and comparison with the CD8alphaalpha homodimer. AB - The crystal structure of a recombinant mouse single chain CD8alphabeta ectodomains at 2.4 A resolution reveals paired immunoglobulin variable region like domains with a striking resemblance to CD8alphaalpha in size, shape, and surface electrostatic potential of complementarity-determining regions (CDR), despite <20% sequence identity between the CD8alpha and CD8beta subunits. Unlike the CD8alpha subunit(s) in the heterodimer or homodimer, the CDR1 loop of CD8beta tilts away from its corresponding CDR2 and CDR3 loops. Consistent with this observation, independent mutational studies reveal that alanine substitutions of residues in the CDR1 loop of CD8beta have no effect on CD8alphabeta coreceptor function, whereas mutations in CD8beta CDR2 and CDR3 loops abolish CD8alphabeta coreceptor activity. The implications of these findings and additional CD8alpha mutational studies for CD8alphabeta- versus CD8alphaalpha-MHCI binding are discussed. PMID- 16356864 TI - Effect of first line therapy including efavirenz and two nucleoside reverse transcriptase inhibitors in HIV-infected children. AB - OBJECTIVE: In an intent-to-treat study, reduction of viral load, increase in CD4 cell count, clinical benefit and adverse reactions were examined in HIV-infected children receiving first line therapy including efavirenz. METHODS: The data of 10 perinatally infected children (median age: 5.8 years) were evaluated during a treatment period of 24 months. Viral load and CD4 cell count were measured every 4 - 8 weeks. Pharmacokinetic evaluations of efavirenz were performed in all patients at study onset. Adverse reactions were reported after obtaining interval history and examination. RESULTS: At base line, median CD4 cell count was 378 cells/microl (21%) and median viral load was 350,000 copies/ml (5.5 log10 copies/ml). After 24 months of treatment, the median viral load reduction was > 3.5 log10 copies/ ml and HIV-1 RNA < 50 copies/ml was found in 8/10 children (80%). Median CD4 cell count increased to 721 cells/microl (24%) after 3 months and was maintained at a level of >1000 cells/microl (> 25%) after 24 months of treatment. Regarding efavirenz levels, C min. values ranged from 845 to 3550 ng/ml (median: 1845 ng/ml) and C max. values from 2380 to 24 200 ng/ ml (median: 3670 ng/ml). The most common adverse effect was a mild skin rash (4/10 children). CNS symptoms were recorded in one patient and no hyperlipidaemia was seen. CONCLUSION: First line therapy with efavirenz and two NRTIs was well tolerated by HIV-1 infected children and the reduction of viral load seems to be similar to single protease inhibitor-containing regimens. PMID- 16356865 TI - Alpha-1-antitrypsin deficiency in children: liver disease is not reflected by low serum levels of alpha-1-antitrypsin - a study on 48 pediatric patients. AB - BACKGROUND: Alpha-1-antitrypsin (alpha1-AT) is an important protease inhibitor. The phenotypes are characterized by a low total serum alpha1-AT or by an abnormal protein accumulating in the hepatocytes. The aim of our study was to examine a correlation of total serum alpha1-AT, phenotype, and liver involvement in pediatric patients. METHODS: 48 patients, deficient for alpha1-AT were included. The phenotypes for alpha1-AT were determined by isoelectric focusing. Liver disease was defined either as elevated transaminases or/and as elevated conjugated bilirubin and gammaGT. Patients were reexamined after a mean interval of 2 years. RESULTS: Homozygous alpha1-AD was found in 12 patients, heterozygous in 24 patients. In 12 children rare variants of alpha1-AD were diagnosed. Serum alpha1-AT levels less than 60% of normal were found in all patients with homozygous, in 37% of patients with heterozygous alpha1-antitrypsin deficiency (alpha1-AD), and in patients with the homozygous variant PiM(palermo). Liver disease was found in 8/12 patients with the phenotype PiZZ and in 15/24 patients with heterozygous alpha1-AD. Three of 4 patients with the phenotype PiMQ0 had severe liver disease despite normal serum levels for alpha1-AT. In 11 patients with heterozygous alpha1-AD liver disease was apparent despite normal serum alpha1-AT levels. In two patients with the variant type Mpalermo serum levels were as low as 11% of normal without any signs of liver disease. CONCLUSIONS: Our data clearly show that in the diagnostic workup of neonatal cholestasis measurement of total serum alpha1-AT does not exclude liver disease due to abnormal alpha1-AT variants. We suggest analysis of alpha1-AT-phenotype by isoelectric focussing in patients with unknown liver disease. Heterozygous or rare variant types might remain undiagnosed by measuring total alpha1-AT only. PMID- 16356866 TI - Management of defects in the groin, thigh, and pelvic region with modified contralateral TRAM/VRAM flaps. AB - In patients with sarcomas, or regional recurrence of the disease, radical resection and radiation therapy is indicated to achieve cure or palliation. As a result of radical surgery, extensive radiation, or infection in the pelvic, groin or thigh region, the development of large pelvic / groin / thigh defects present a difficult surgical problem. Musculocutaneous deep epigastric island-flaps, as a modification of the extended deep inferior epigastric flap, described by Taylor et al. in 1983, are an attractive option for a successful reconstruction for this defect localisation. Two technical modifications of the contralateral extended deep inferior epigastric island-flap permitted an adequate tailoring to a defect in the posterior lateral pelvic or groin-thigh region, and avoided the drawbacks of other loco-regional or microsurgical flaps in previously irradiated fields. The skin island reached the posterior lateral pelvic region, groin or thigh. The muscular portion of this flap was suited to fill the soft tissue defects in these critical areas. These so modified flaps represent an instrument, which in selected cases may be an interesting alternative tool for closure of complex defects. No significant functional impairment secondary to the flap procedures was noted in our patients. PMID- 16356867 TI - Adverse events of sutures: possible interactions of biomaterials? AB - Absorbable sutures are in use for more than 30 years. Tissue reactions which might be associated with suture material have rarely been reported in the past. After a long period without complications caused by suture material we observed 12 cases of unexpected tissue reactions after clean operations. Our patients 3-8 weeks after uneventful elective clean operations (varicose vein, hernia, benign soft tissue tumor) had unexpected tissue reactions (inflammation, granuloma, extrusion, fistula, abscess) in the vicinity of Vicryl, suture material (8 cases with Vicryl, 4 cases with Vicryl plus. After removal of the suture material and the granulomatous tissue wounds healed without any further disturbance. These tissue reactions have been observed in patients with subcuticular sutures as well as in patients with deeper located vein ligatures. It is well known that next to patient-associated and surgeon-related factors biomaterials might have an impact on postoperative inflammatory process and healing. We use Vicryl, suture material for ambulatory surgery since 1999 and did not see complications for a long period up to now. 11 of the patients were observed within several weeks in summer 2005, whereas only one patient has been observed in the year 2004. All 11 patients observed in 2005 had a combination of Vicryl/ Vicryl plus suture material in deep/subcutaneous and Dermabond glue for skin closure. We do not know the cause for this change. For clarification evaluation of the tissue reactions of these biomaterials including possible interactions or combined reactions should be done. PMID- 16356868 TI - Therapy of focal viral encephalitis in children with aciclovir and recombinant beta-interferon - results of a placebo-controlled multicenter study. AB - Focal viral encephalitis in childhood is a rare but life-threatening disease. Animal experiments and case reports suggest a positive effect of an additional therapy with interferon-beta on the course of the disease. Therefore, we initiated a prospective, double-blind placebo-controlled study to investigate the benefit of a combination therapy of Aciclovir (ACV) and recombinant interferon beta (rIFN-beta) in juvenile focal viral encephalitis. - Initial inclusion criterium was suspicion of focal viral encephalitis. Diagnosis was proven by demonstration of characteristic focal lesions in cerebral imaging or virological evidence of HSV in cerebrospinal fluid. Patients were treated with ACV plus rIFN beta or ACV plus placebo. Neurological outcome was determined 21 days and 3 months after onset of the disease. - Initially 59 patients were enrolled in the study. Encephalitis was proven in 14 patients (7 ACV + rIFN-beta, 7 ACV + placebo). The study groups were balanced in terms of important prognostic criteria. 10 patients (5 ACV + rIFN-beta, 5 ACV + placebo) were cured or had slight defects, 4 patients (2 ACV + rIFN-beta, 2 ACV + placebo) showed moderate to severe defects. There was no significant difference in favour of the additive therapy with rIFN-beta. PMID- 16356869 TI - Spontaneous regression of retinal angiomatous lesions in v. Hippel-Lindau disease (VHL). AB - BACKGROUND: Very little documentation of spontaneous regression of an angiomatous retinal lesion in v. Hippel-Lindau disease (VHL) exists. It is commonly believed that a spontaneous change of hemangiomas into fibrotic lesions occurred. PATIENTS/METHODS: Follow-up examinations of four patients with VHL in the Freiburg VHL study were carried out. RESULTS: A 16-year-old girl revealed a vascular lesion at the border of the optic disc. Control examination nine years later revealed complete spontaneous regression of the retinal vascular changes. A slight retinal vascular change at the superior border in her right eye was found in a 36-year-old woman. A control examination 20 years later revealed regression of the lesion. A 41-year-old woman showed in the retinal periphery a small fibrotic white hemangioma with a pigmented feeder vessel as sign of spontaneous tumor regression. A 12-year-old boy had a retinal microaneurysm inferior to the optic disc that disappeared several years later. CONCLUSION: Documentations of spontaneous regression of minor angiomatous retinal lesions in VHL exist. Such vascular changes are rare. Every retinal lesion should be controlled by follow-up examination and documentation. In case of retinal lesion growth, treatment is necessary. PMID- 16356870 TI - The leukocyte count predicts the efficacy of treatment with azathioprine in inflammatory bowel disease. AB - INTRODUCTION: Azathioprine has variable efficacy in inflammatory bowel disease. Previous studies suggested that either neutropenia, an increase in the mean corpuscular volume, the assessment of thiopurine methyl-transferase activity or erythrocyte 6-thioguanine values might predict the treatment response. However, due to the conflicting results of the preceding studies there are yet no established laboratory values which allow an estimation of the clinical response. PATIENTS AND METHODS: 45 patients with Crohn's disease and 39 patients with ulcerative colitis were enrolled in this retrospective evaluation. After a minimum of six months therapy with azathioprine patients in remission were compared with those who did not achieve a stable remission with respect to the number of leucocytes, lymphocytes, neutrophil granulocytes and the mean corpuscular volume. RESULTS: Patients who went into remission during treatment with azathioprine displayed significantly lower leukocyte counts if compared to patients who were not in remission (p = 0.004 in Crohn's disease and 0.003 in ulcerative colitis). A similar tendency was also observed with respect to the granulocyte count (p = 0.007 in Crohn's disease and 0.004 in ulcerative colitis). The mean corpuscular volume did not correlate with the response to purine analogues. DISCUSSION: The absolute leukocyte count and the percentage of granulocytes seem to predict the response to purine analogues in inflammatory bowel disease and possibly offers a feasible and cost effective diagnostic tool for the assessment of therapeutic efficacy. Subsequent preferably prospective studies should aim to define the optimal cut-off value for the leukocyte count. PMID- 16356871 TI - Disease course after liver transplantation for hepatocellular carcinoma in patients with complete tumor necrosis in liver explants after performance of bridging treatments. AB - AIM: To study the disease course of patients with hepatocellular carcinoma (HCC) showing complete tumor necrosis in their liver explants after undergoing bridging treatments followed by liver transplantation (LTx). PATIENTS AND METHODS: We evaluated data corresponding to 10 patients with liver cirrhosis undergoing bridging treatments for HCC prior to LTx. In all cases there was complete tumor necrosis in the explanted livers. RESULTS: There were 8 men and 2 women. Percutaneous radiofrequency ablation (RFA) was performed under computed tomographic guidance in 4 patients. The remaining 6 patients underwent transarterial chemoembolization (TACE). Five of them received one session of TACE, while the remaining one received a series of 4 sessions prior to LTx. Six patients had solitary nodules with a median diameter of 3.5 cm (range 2.5-4.2 cm). Four of them underwent RFA. Segmental tumor chemoembolization was performed in 2 patients. The remaining 4 patients had 2 tumors each with a median total diameter of 4.4cm (range 4.2-6.0 cm) prior to TACE. They underwent bilobar hepatic chemoembolization, which under staged the tumors prior to live donor liver transplantation (LDLTx). Six patients underwent deceased donor orthotopic liver transplantation. LDLTx was performed in 4 patients. Median waiting time to LTx was 53 days. All patients are alive without recurrence after a median follow up of 19 months. CONCLUSION: Achievement of 100% tumor necrosis by means of bridging treatments followed by LTx for HCC is characterized by a very low recurrence rate and should receive further consideration and study. PMID- 16356872 TI - Participation of tyrosine kinase and phospholipase Cgamma in isradipine-induced proliferation of cultured human gingival fibroblasts. AB - Some kinds of drugs such as calcium (Ca(2+)) channel antagonists, antiepileptics and immunosuppressants cause gingival overgrowth as a side effect, the mechanism of which is still unclear. We have examined the effects of isradipine, one of the dihydropyridine-derivative Ca(2+) channel antagonists, on cultured human gingival fibroblast Gin-1 cells. In the present study, to elucidate the mechanism by which isradipine causes gingival overgrowth, we examined whether tyrosine kinase (TK) and phopholipase Cgamma (PLCgamma) are involved in the isradipine-induced proliferation of gingival fibroblasts. Herbimycin A (1 microM) remarkably inhibited the isradipime (10 microM)-induced proliferation. Both U73122 (5 microM), a PLCgamma inhibitor, and xestospongin C (5 microM), an antagonist of a receptor of inositol 1,4,5-trisphosphate in Ca(2+) stores, significantly reduced the [Ca(2+)]i raised by isradipine (10 microM). Thus, the findings obtained here indicate that TK and PLCgamma are closely involved in the isradipine-induced [Ca(2+)]i rise to elicit gingival overgrowth. PMID- 16356873 TI - Immunohistochemical characteristics of developing mandibular angle in fetal mice. AB - Mouse mandibular angle development started as a coagulation of mesemchymal cells on the 15(th) fetal day. On the 16(th )fetal day, cells of the central portion of the cell coagulation showed metachromasia to toluidine blue, and type 2 collagen positive chondrocytes were immunohistochemically detected. After the 17(th) fetal day, cartilaginous osteogenesis occurred with invasion of capillaries. At the same stage, membranous (perichondral) ossification occurred in the periphery of the chondrocyte mass. These proliferating chondrocytes showed positive reactions to type 2 collagen, type 1 collagen and osteopontin. These results suggest that the characteristics of mandibular angular cartilage are slightly different from those of normal physiological articular cartilage. PMID- 16356875 TI - What women want and what their doctors need--how do women evaluate the quality of contraceptive methods and products introduced by their gynaecologist? A different perspective. AB - Is a doctor really aware of women's expectations and needs when choosing their contraceptive? When faced with making a decision about which contraceptive to chose, women often turn to their doctor for information and guidance. This is particularly the case with oral hormonal contraceptives. Nowadays women often demand more than just a pill that will provide reliable contraception--they expect additional benefits such as good cycle control, improvement in dysmenorrhoea and if possible a beneficial effect on hair and skin. The increased choice of available oral contraceptives places additional pressure on doctors to fully understand the products they are prescribing, in terms of both the physiological and the psychological effects they may have on their patients. Clinical guidelines need to be in place in order to manage any day-to-day problems that may arise. In addition doctors need to re-evaluate their relationship with their patients in order to provide what women really want. PMID- 16356876 TI - Chlormadinone acetate (CMA) in oral contraception--a new opportunity. AB - Chlormadinone acetate (CMA) is a derivative of naturally secreted progesterone that shows high affinity and activity at the progesterone receptor. It has an anti-estrogenic effect and, in contrast to natural progesterone, shows moderate anti-androgenic properties. CMA acts by blocking androgen receptors in target organs and by reducing the activity of skin 5alpha-reductase. It suppresses gonadotropin secretion and thereby reduces ovarian and adrenal androgen production. CMA shows high contraceptive efficacy by inhibiting ovulation due to its ability to suppress or disrupt endogenous gonadotropin secretion and, by this, inhibits follicular growth and maturation. In addition, it suppresses endometrial thickness and increases the viscosity of cervical mucus. Pharmacokinetic studies have shown rapid and almost complete absorption after oral administration, and CMA is being bound to albumin rather than SHBG (Sex Hormone-Binding-Globulin). Multiple dosing studies have demonstrated that steady state is reached by day 7 after oral administration with peak plasma concentrations in the region of 2 ng/ml. After a single dose of CMA the half-life time is around 34 hours and after multiple dose administration approximately 38 hours. Safety studies have indicated that CMA has no clinically relevant effect on a wide range of metabolic parameters in normal subjects. Further studies in groups at high thromboembolic risk have shown that CMA alone produces a relative risk of 0.8 which is not considered significant. These results indicated the potential for CMA to be combined with ethinylestradiol in an oral contraceptive which provides highly effective contraception and excellent cycle control. PMID- 16356877 TI - Belara--a reliable oral contraceptive with additional benefits for health and efficacy in dysmenorrhoea. AB - Although modern oral contraceptives are safe and have few side-effects, compliance towards them is sometimes less than ideal for various reasons. Compliance, however, can only be achieved when the contraceptive method is accepted by the users, that is, when it is adapted to their individual needs. Consisting of a combination of 2 mg chlormadinone acetate and 0.03 mg ethinylestradiol, Belara is a modern oral hormonal contraceptive with an unadjusted Pearl index of 0.44 (95% CI, 0.2-0.8) and an adjusted one of 0.04 (95% CI, 0.002-0.2). Its compliance rate in clinical use has been shown to be above 90%. This good acceptance is a consequence of the low rate of intermenstrual bleeding (about 8% up to the 3(rd) cycle and below 2% from the 12(th) cycle); its high cycle stability (in approximately 98% from the 6(th) cycle); the good weight stability (weight is unchanged in about 84% from the 12(th) cycle); and finally the very low rate of side-effects (below 2% after 12 cycles). In addition, a number of other benefits of using Belara also contribute to this good compliance rate. These include almost 70% improvement or complete remission of increased seborrhoea after 12 months, almost 90% improvement or cure of acne after 12 months, and improvement or remission of dysmenorrhoea after 12 months in 79% of cases. After 4 months, improvement or remission of dysmenorrhoea associated with the use of another ovulation inhibitor was seen in more than 90% of cases after switching to Belara. In conclusion, besides being an effective, modern oral hormonal contraceptive Belara offers a considerable range of additional benefits for a range of symptoms, including primary dysmenorrhea and acne. PMID- 16356878 TI - Belara--proven benefits in daily practice. AB - Today, a contraceptive method is available to suit nearly every type of woman, every age and all preferences and expectations. All that seems to remain for users is to look for the right product to satisfy their personal requirements. The physician takes on the role of the adviser, responsible mainly for errors of judgement and undesirable effects. The choice of the suitable contraceptive depends on three factors: the patient profile, the profile of the method used and the user's life situation. In selecting the method of contraception, statistical measures such as the Pearl Index, rate of adverse events, risks and health benefits as well as the pharmacological profile, resulting intake modality and potential interactions should be considered. The patient profile includes both subjective wishes and standards of value relevant for world view, family planning and psychological well-being, as well as objective parameters such as age, BMI, medical history and the woman's sexual behaviour. Evaluation of these parameters by the physician is a major component of successful contraceptive counselling. Belara is a new oral contraceptive on the European market based on a monophasic combination of 2 mg chlormadinone acetate and 0.03 mg ethinylestradiol. As well as high contraceptive efficacy and a low rate of side effects, Belara features an outstanding safety profile due to its almost complete absence of mineralocorticoid and glucocorticoid action and its absent impact on hepatic metabolism. In daily practice, Belara exhibits mild antiandrogenic activity which also makes it suitable for users with antiandrogen-induced seborrhoea and moderate acne. Symptoms of PMS or unspecific dysmenorrhea and menstrual irregularities can also be alleviated or completely eliminated by taking Belara. Belara use has not been associated with any significant weight gain. In daily practice, Belara is suitable for every woman of every age without specific risk factors requiring safe contraception. Belara also has considerable additional health benefits that should also be considered when choosing a suitable contraceptive. PMID- 16356879 TI - Method development and HPLC analysis of retail foods and beverages for copper chlorophyll (E141[i]) and chlorophyllin (E141[ii]) food colouring materials. AB - An analytical method using high performance liquid chromatography with photodiode array and fluorescence detection has been developed and applied to the determination of the food colour additives copper chlorophylls and copper chlorophyllins (E141[i] and [ii]) in foods and beverages. The analytical procedures from previously reported methods have been refined to cover a range of food colour formulations and retail foods. The method was single-laboratory validated. Recoveries of the polar copper chlorophyllins from spiked samples (at 14.5 mg/kg in all but one case) were in the range 79-109%, except for jelly sweets (49%). Recoveries of relatively non-polar copper chlorophylls were in the range 77-107% (except for 'made' jelly at 50%). The %RSD for recoveries was generally below 12%. Quantitative estimates of the total copper chlorophyll/chlorophyllin content of a small range of food commodities are reported, based on the use of trisodium copper chlorophyllin as a surrogate standard. The majority of E141-containing foods and colour formulations analysed exhibited a multiplicity of components due to the various extraction and purification processes that are used to obtain these colour additives. This was confounded by the presence of overwhelming amounts of native chlorophylls in certain samples (e.g. mint sauce). Food commodities containing significant amounts of emulsifiers (i.e. ice cream), gelatine or fats were problematic during extraction hence further development of extraction regimes is desirable for such products. All of the samples analysed with added E141, had estimated total copper chlorophyllin contents of below 15 mg/kg (range 0.7-13.0). PMID- 16356880 TI - Estimated daily intakes of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ) antioxidants in Korea. AB - The study was conducted to establish the estimated daily intake (EDI) of antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ) in Korea. The EDIs were obtained from two sources. One of the estimations was based on the analytical determination of BHA, BHT and TBHQ in 12 food categories (ten food categories for TBHQ) and on individual dietary intake data obtained from the National Health and Nutrition Survey in 1998 (n=11 525, age > 1 year). The other EDIs of BHA, BHT and TBHQ were based on the maximum permitted levels specified in national food standards in Korea and on individual dietary intake data obtained from the National Health and Nutrition Survey in 1998 (n=11 525, age > 1 year). To establish the EDIs based on the analytical determination and on individual dietary intake data, 133 food samples in 12 food categories were selected from the foods considered to be representative sources of BHA, BHT and TBHQ in the Korean diet. Selected samples were analysed by GC with FID. BHA was not detected in any of the samples analysed. BHT and TBHQ were detected in the samples, but the levels were significantly lower than their maximum limits. The EDIs1 of BHT, and TBHQ for average consumers were 0.0156(-3), and 0.0012(-3) mg kg(-1) body weight bw day( 1) and as a proportion of the ADI were 0.0052 and 0.0002%, respectively. For 95th percentile consumers, the EDIs of BHT and TBHQ were 0.0080 and 0.0006 mg kg(-1) bw day(-1), and as a proportion of the ADI were 2.67 and 0.09%, respectively. EDIs for BHA, BHT and TBHQ based on the maximum permitted levels and on individual dietary intake data were 0.04, 0.04 and 0.04 mg kg(-1) bw day(-1), respectively. The EDIs of BHA, BHT and TBHQ for average consumers ranged from 6.00 to 14.42% of the ADI of each antioxidant. According to these results, the EDIs of BHA, BHT and TBHQ in Korea were significantly lower than ADI of these antioxidants established by the JECFA. PMID- 16356881 TI - Determination of 3-chloropropane-1,2-diol as its 1,3-dioxolane derivative at the microg kg-1 level: application to a wide range of foods. AB - This paper reports the application of a GC/MS method for the quantification of 3 chloropropane-1,2-diol (3-MCPD) at low microg kg-1 levels through the determination of its 1,3-dioxolane derivative to a wide range of foodstuffs. The proposed protocol is based on the methods generally performed in control laboratories. The two main stages of the method - the solid-phase extraction and the purification of the derivatives - have been optimized. The within-laboratory reproducibility meets the official performance criteria for verifying 3-MCPD at the 20 microg kg(-1) limit stipulated by the European Community for soy sauce. The limit of quantification was below 5 microg kg(-1) for all the foodstuffs analysed. This method offers a valuable alternative to the draft CEN European Standard: instead of diethyl ether, much safer ethyl acetate is used; derivatization is more selective; and reagents are common stable chemicals. This method was successfully applied to toasted bread, savoury crackers, cheese, soups, including vegetable-based soups, meat products such as salami, vegetable oils, sauces, soy sauce and related products. Upon checking the method performance in the case of vegetable oils, the unexpected presence of monobromopropanediols was detected. PMID- 16356882 TI - Analysis of furan in foods. Is headspace sampling a fit-for-purpose technique? AB - Headspace GC-MS has been optimized for the determination of furan in foods. The conditions of sample preparation, headspace sampling and GC separation were optimized to enhance sensitivity during GC-MS analysis. Green coffee was used to prepare a matrix matched calibration curve for furan. However, it was unexpectedly found that a green coffee sample was not blank. GC-MS analysis performed after equilibration for 30 min at 40 degrees C showed the presence of 4.2 ng/g furan in green coffee. In order to understand whether furan was naturally present or formed during headspace sampling, green coffee was investigated in time-dependent manner at headspace equilibration temperatures of 40 and 70 degrees C. It was observed that furan response continued to increase in a way similar to first order formation kinetics. The same behavior was found for freshly squeezed tomato and orange juices leading to the suspicion of furan formation during headspace equilibration. It is concluded that a matrix matched calibration for each particular food matrix is necessary to compensate for furan formation during headspace sampling, and thus, to quantify furan more accurately. PMID- 16356883 TI - Evaluation of the nitrate content in leaf vegetables produced through different agricultural systems. AB - The nitrate content of leafy vegetables (watercress, lettuce and arugula) produced by different agricultural systems (conventional, organic and hydroponic) was determined. The daily nitrate intake from the consumption of these crop species by the average Brazilian consumer was also estimated. Sampling was carried out between June 2001 to February 2003 in Campinas, Sao Paulo State, Brazil. Nitrate was extracted from the samples using the procedure recommended by the AOAC. Flow injection analysis with spectrophotometric detection at 460 nm was used for nitrate determination through the ternary complex FeSCNNO+. For lettuce and arugula, the average nitrate content varied (p < 0.05) between the three agricultural systems with the nitrate level in the crops produced by the organic system being lower than in the conventional system that, in turn, was lower than in the hydroponic system. For watercress, no difference (p < 0.05) was found between the organic and hydroponic samples, both having higher nitrate contents (p < 0.05) than conventionally cultivated samples. The nitrate content for each crop species varied among producers, between different parts of the plant and in relation to the season. The estimated daily nitrate intake, calculated from the consumption of the crops produced by the hydroponic system, represented 29% of the acceptable daily intake established for this ion. PMID- 16356884 TI - Easy determination of the addition of soybean proteins to heat-processed meat products prepared with turkey meat or pork-turkey meat blends that could also contain milk proteins. AB - The addition of non-meat proteins to processed meat products is limited by regulations. Therefore, this work has investigated the determination of added soybean proteins in commercial heat-processed meat products prepared with turkey meat or pork-turkey meat blends that could also contain milk proteins. The method consisted of extracting proteins from the meat products in a Tris-HCl buffer (pH 8) and analysing the extract by high-performance liquid chromatography with a linear gradient water-acetonitrile containing 0.05% (v/v) TFA. This method enabled the detection and quantitation of up to 0.08 and 0.28% (w/w), respectively, of soybean proteins (related to 6 g initial product) in these products. Satisfactory precision and recovery data were established. Accuracy was evaluated by a comparison of soybean protein contents determined by the proposed method and the existing AOAC official method based on an enzyme-linked immunosorbent assay (ELISA) from which no statistically significant differences were observed. PMID- 16356885 TI - Justifying the need to prescribe limits for toxic metal contaminants in food grade silver foils. AB - The use of silver foils in various food preparations is a common practice in Middle Eastern and South East Asian countries. The FAO/WHO Joint Expert Committee on Food Additives (JECFA) has included silver in the list of food additives, but specifications were not prepared. Indian food legislation has included food-grade silver foil and laid down a purity requirement of 99.9%. This leaves an unspecified margin of 0.1% or 1000 microg g(-1) for contaminants. Therefore, a study to investigate the levels of metallic contaminants in food-grade silver foil was undertaken. Of 178 foils analysed, 161 (90%) contained silver, whilst 10% were fraudulently made up of aluminium. In the case of silver foils, 46% of the samples adhered to the desired purity requirement of 99.9%, while 54% had a lower silver content. Copper was present in 86.3% of the silver foils, while chromium, nickel and lead contamination was found in over 54% of samples. Cadmium levels were detected in 28% of the silver foils and manganese was present in 6.8% of samples. In silver foils showing metal contaminants, average levels were found for nickel (487 microg g(-1)), lead (301 microg g(-1)), copper (324 microg g( 1)), chromium (83 microg g(-1)), cadmium (97 microg g(-1)) and manganese (43 microg g(-1)), which being appreciable justify the need to prescribe limits for some metals in food-grade silver foil as well as for silver powder used in confectioneries and medicinal preparations. The work reported here should encourage manufacturers to use high-purity raw materials and take suitable precautions to reduce unwarranted exposure of consumers to toxic metal contaminants. PMID- 16356886 TI - Quality and safety assessment of ginseng extracts by determination of the contents of pesticides and metals. AB - Ginseng extracts are available as ingredients for improving energy and vitality and can be used in functional foods and as flavouring ingredients. A survey was been performed to determine the content of pesticides and toxic metals in ginseng extracts. Forty-seven samples from 20 suppliers, including both Panax ginseng C. A. Meyer (Asian ginseng) and P. quinquefolius (American ginseng) species, were analysed for arsenic content and for the following metals: aluminium, molybdenum, chromium, copper, magnesium, zinc, cadmium, mercury and lead, while pesticide residues were analysed in 30 samples from 17 suppliers. The results showed that 24 samples (80%) contained pesticides above the detection limit and 13 samples (43%) did not comply with the maximum residue limits (MRL) for total quintozene, hexachlorobenzene, total hexachlorocyclohexane, lindane, total heptachlor, e chlorpyrifos and folpet, imposed for botanical extracts. Total quintozene, hexachlorobenzene, total hexachlorocyclohexane and lindane were present in all contaminated samples and exceeded the MRL in eleven samples, with levels up to 55 and 30 times their respective MRL. Cadmium (<0.05-259 microg kg(-1)), mercury (<0.3-72 microg kg(-1)), lead (3-2710 microg kg(-1)) and arsenic (<0.3-918 microg kg(-1)) were present in most samples at concentrations lower than the MRL imposed for flavouring substances. Among the other elements, aluminium (0.3-1068 mg kg( 1)) was the most abundant. PMID- 16356887 TI - Baby food production chain: pesticide residues in fresh apples and products. AB - During 3 years of a monitoring programme, 522 samples of fresh apples, six brands of fruit purees and various types of fruit baby food prepared from these materials were analysed. Each sample was examined for the presence of 86 GC amenable pesticide residues. The reporting limits of the procedure employed for sample analyses were in the range 0.003-0.01 mg kg(-1). Pesticide residues were detected in 59.5% of the samples of fresh apples. However, maximum residue levels (European Union MRLs) were exceeded only in 1.4% of samples. The levels of residues in 'positive' fruit purees were substantially lower, overall with residues detected in 33% of samples. Fruit baby food represented the commodity with the lowest incidence of residues being detected in only 16% of samples. The 0.01 mg kg(-1) MRL was exceeded in 9% of these products. Multiple residues were found in 25% of fresh apples and in 10% of fruit purees. None of fruit baby food samples contained more than a single residue. Organophosphorus insecticides and fungicides representing phtalimides, sulphamides and dicarboximides were the most frequently found residues. To obtain more knowledge on the fate of residues during fruit baby food production, processing experiments employing apples with incurred residues (fenitrothion, phosalone and tolylfluanid) were conducted. Washing of apples did not significantly reduce the content of pesticides. Steam boiling followed by removal of peels/stems was identified as the most efficient steps in terms of residues decrease (phosalone) or complete elimination (fenitrothion and tolylfluanid). PMID- 16356888 TI - Studies on the affinity chromatography purification of anti-patulin polyclonal antibodies by enzyme linked immunosorbent assay and electrophoresis. AB - Patulin is a mycotoxin produced by fungal species that frequently grow on fruit and vegetables. It presents risks, particularly for children consuming compotes and fruit juices. Thus, it is important to have methods such as immunoassays to screen a large number of samples. In the relevant literature, previous studies on the production of antibodies against patulin derivatives described qualitative tests for a patulin derivative or showed slight responses. The present study reinvestigated the production of polyclonal antibodies against patulin and their purification since crude antiserum could react non-specifically in immunoassays. Patulin-hemiglutarate was synthesized and conjugated to bovine serum albumin as the immunogen for the immunization of five New Zealand white rabbits. The immunoglobulin G (IgG) fraction was isolated twice by affinity chromatography using Sepharose-LS gel and recombinant G-protein. Classic affinity chromatography using Sepharose-LS gel was unable to eliminate serum albumin from the IgG fraction and the use of recombinant G-protein was efficient to isolate the purified IgG. Titres and specificity were determined by indirect competitive enzyme-linked immunosorbent assay. Patulin-hemiglutarate-ovalbumin gave complete displacement, while patulin displaced 30% of bound antibodies. Thus, a fraction of the antibodies are specific for free patulin. The non-specific binding increased with patulin concentrations. The electrophilic properties of patulin might also induce intermolecular cross-links in vitro that hinder the possibility of responses displacement when free patulin is used. PMID- 16356889 TI - Survey of aflatoxins in beer sold in Canada. AB - Between March 1998 and March 2002, 304 samples of domestic (Canadian) and imported beers from 36 countries were picked up for the determination of aflatoxins B1, B2, G1 and G2. Twelve samples were positive with aflatoxins greater than the limit of quantitation (LOQ) (aflatoxin B1, 4.4 ng l(-1); aflatoxin B2, 3.4 ng l(-1); aflatoxin G1, 11.2 ng l(-1); and aflatoxin G2, 6.2 ng l(-1)). Five samples from Mexico, two samples from Spain and one from Portugal contained aflatoxin B1. Four samples from India contained aflatoxins B1 and B2. The remaining samples contained less than the LOQ for aflatoxins B1, B2, G1 and G2. The analytical method for this survey was based on that of Scott and Lawrence (Scott PM, Lawrence GA. 1997. Determination of aflatoxins in beer. Journal of AOAC International 80:1229-1234.). Aflatoxins B1, B2, G1 and G2 were determined at parts per trillion (ng l(-1)) levels in beer by immunoaffinity column cleanup followed by derivatization with trifluoroacetic acid and reversed-phase liquid chromatography with fluorescence detection. PMID- 16356890 TI - Incidence of toxigenic fungi and ochratoxin A in dried fruits sold in Brazil. AB - A total of 117 dried fruit samples (black sultanas, white sultanas, dates, dried plums, dried figs and apricots) from different origins were analysed both for toxigenic fungi and for the presence of ochratoxin A. Amongst the fungi found, Aspergillus niger was predominant, with 406 isolates, of which 15% were ochratoxin A producers. They were followed by A. ochraceus, with 15 isolates and 87% ochratoxigenics, and A. carbonarius, with only five isolates of which 60% were ochratoxin A producers. The average infection rates for A. niger in black sultanas, plums, figs, dates and white sultanas were 22.0, 8.0, 4.0, 1.5 and 0.5%, respectively. The apricot samples were not contaminated by any fungi or ochratoxin A. Black sultana and dried figs contained the highest contamination with ochratoxin A, with 33 and 26.3% of the samples containing more than 5 microg kg(-1) respectively, while all the white sultanas, dates and plums had no sample that exceeded this limit. PMID- 16356891 TI - Volatile and non-volatile radiolysis products in irradiated multilayer coextruded food-packaging films containing a buried layer of recycled low-density polyethylene. AB - The effects of gamma-irradiation (5-60 kGy) on radiolysis products and sensory changes of experimental five-layer food-packaging films were determined. Films contained a middle buried layer of recycled low-density polyethylene (LDPE) comprising 25-50% by weight (bw) of the multilayer structure. Respective films containing 100% virgin LDPE as the buried layer were used as controls. Under realistic polymer/food simulant contact conditions during irradiation, a large number of primary and secondary radiolysis products (hydrocarbons, aldehydes, ketones, alcohols, carboxylic acids) were produced. These compounds were detected in the food simulant after contact with all films tested, even at the lower absorbed doses of 5 and 10 kGy (approved doses for food preservation). The type and concentration of radiolysis products increased progressively with increasing dose. Generally, there were no significant differences in radiolysis products between samples containing a buried layer of recycled LDPE and those containing virgin LDPE (all absorbed doses), indicating the good barrier properties of external virgin polymer layers. Volatile and non-volatile compounds produced during irradiation affected the sensory properties of potable water after contact with packaging films. Taste transfer to water was observed mainly at higher doses and was more noticeable for multilayer structures containing recycled LDPE, even though differences were slight. PMID- 16356892 TI - Compositional GC-FID analysis of the additives to PVC, focusing on the gaskets of lids for glass jars. AB - A gas chromatographic (FID) method is described which aims at the quantitative compositional analysis of the additives in plasticized PVC, particularly the plastisols used as gaskets for lids of glass jars. An extract of the PVC is analysed directly as well as after transesterification to ethyl esters. Transesterification enables the analysis of epoxidized soya bean and linseed oil (ESBO and ELO) as well as polyadipates. For most other additives, the shifts in the chromatogram resulting from transesterification is used to confirm the identifications made by direct analysis. In the gaskets of 69 lids from the European market used for packaging oily foods, a broad variety of plastisol compositions was found, many or possibly all of which do not comply with legal requirements. In 62% of these lids, ESBO was the principal plasticizer, whereas in 25% a phthalate had been used. PMID- 16356894 TI - Behaviors of children following a randomized controlled treatment program for their abused mothers. AB - OBJECTIVE: To determine if a treatment program offered to abused mothers positively affects the behaviors of their children. METHODS: A randomized, two arm, clinical trial was used to measure child behavior at 6, 12, 18, and 24 months following the application of two levels of abuse treatment services to abused mothers: (1) abuse assessment and receipt of a wallet-size referral card, or (2) abuse assessment, receipt of a wallet-size referral card, and nurse case management sessions. The setting was public primary care clinics. The participants were 233 women who reported physical or sexual abuse within the preceding 12 months, and who had at least one child, ages 18 months to 18 years, living with them. Outcome measures were scores on the Child Behavior Checklist (CBCL) at baseline, 6, 12, 18, and 24 months. CBCL scores for a clinically referred sample of children served as a comparison group. RESULTS: All children improved significantly (p < .001) on CBCL scores from intake to 24 months, regardless of which treatment protocol their mother received. By 24 months, the majority of children and adolescents had scores significantly less than the referred norms. Children ages 18 months to 5 years showed the most improvement and teenagers showed the least improvement. CONCLUSIONS: Disclosure of abuse, such as that which happens during abuse assessment, was associated with the same improvement in child behavior scores as a nurse case management intervention. Routine abuse assessment and referral have the potential to positively improve the behavioral functioning of children exposed to domestic violence. PMID- 16356895 TI - Assessing the feeding/eating interaction as a context for the development of social competence in toddlers. AB - Social competence likely develops through the reciprocal nature of mother-child interactions. Interactions around food provide the young child with consistent and predictable social experiences with the mother, which may establish templates for interactive patterns with others. The Toddler Snack Scale (TSS) assesses the pattern of toddler social behaviors in relation to maternal behaviors during an eating episode. Scale reliability was examined with a sample of 126 dyads at the child's ages of 12, 24, and 36 months. Significant associations were found between TSS classifications and concurrent measures assessed with the Nursing Child Assessment Teaching Scale (NCATS), the Control-Autonomy Balance Scale (CABS), and the Adaptive Social Behavioral Inventory (ASBI). Child temperament contributed to child expressions of social competence, but not to the assignment of maternal or child interaction styles. The scale identifies salient areas for parent-child assessment and intervention throughout infancy and toddlerhood. PMID- 16356896 TI - Living on the streets in Canada: a feminist narrative study of girls and young women. AB - Homelessness affects thousands of girls and young women in Canada. Terms that are commonly used to describe the homeless include lazy, mentally ill, middle aged, and male. The reality is that homelessness is not limited to a particular age, gender, or ethnocultural group, or to individuals of any single intellectual ranking. As a result of the prevailing stereotypes, little research has been conducted on homelessness among adolescent females, making it difficult to capture an accurate and comprehensive picture of the full scope of the problem. The purposes of this feminist narrative study were to (1) explore the intersections between homelessness and health among adolescent girls, with particular attention to the influence of contextual factors such as violence, gender, and poverty; and (2) to examine how these young women access health care, barriers they encounter, and factors that contribute to their health promotion. The sample consisted of ten females, ages 16-21, who were interviewed either individually or in small groups. Consistent with feminist research approaches, the interviews were conducted in an interactive manner in order to encourage critical reflection and dialogue. Data analysis consisted of a thematic analysis of the participants' experiences. Findings revealed that most girls had fled from difficult, and at times dangerous, situations at home to lives on the street that brought a new set of challenges, including a multitude of health problems and exposure to violence, chronic poverty, and discrimination. Many barriers to effective health care were described. In this article, the findings of the research will be presented and implications for health and social service providers will be addressed, including recommendations for programming and policy. PMID- 16356899 TI - A mathematical model linking tree sap flow dynamics to daily stem diameter fluctuations and radial stem growth. AB - To date, models for simulating sap flow dynamics in individual trees with a direct link to stem diameter variation include only the diameter fluctuation driven by a change in stem water storage. This paper reports results obtained with a comprehensive flow and storage model using whole-tree leaf transpiration as the only input variable. The model includes radial stem growth based on Lockhart's equation for irreversible cell expansion. It was demonstrated that including growth is essential to obtaining good simulation results. To model sap flow dynamics, capacitance of storage tissues was assumed either constant (i.e., electrical analogue approach) or variable and dependent on the water content of the respective storage tissue (i.e., hydraulic system approach). These approaches resulted in different shapes for the desorption curve used to calculate the capacitance of storage tissues. Comparison of these methods allowed detection of specific differences in model simulation of sap flow at the stem base (F(stem)) and stem diameter variation (D). Sensitivity analysis was performed to select a limited subset of identifiable parameters driving most of the variability in model predictions of F(stem) and D Both the electrical analogue and the hydraulic system approach for the flow and storage model were successfully calibrated and validated for the case of a young beech tree (Fagus sylvatica L.). Use of an objective model selection criterion revealed that the flow and storage model based on the electrical analogue approach yielded better predictions. PMID- 16356897 TI - Parents' use of nonmedical support services in the neonatal intensive care unit. AB - Parents frequently identify the need for support while their infant is in the Neonatal Intensive Care Unit (NICU), however, they may simultaneously distance themselves from traditional family and friend support. Recognizing this, many NICUs provide additional nonmedical support services such as social workers, chaplains/religious counselors, and support groups. This article, part of a larger research study, suggests an inverse relationship between social support and the use of supportive services. In addition, parents in this study appear to use support services less often than would be anticipated based on their reports of utility. Suggestions are provided to potentially improve desirability/accessibility of these services. PMID- 16356900 TI - Transpiration-induced axial and radial tension gradients in trunks of Douglas-fir trees. AB - We determined the axial and radial xylem tension gradients in trunks of young Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees. Axial specific conductivity (k(s-a)) and sap flux density (Js) were measured at four consecutive depths within the sapwood at a stem height of 1 m. By definition, at a given position in the bole, Js is a function not only of k(s-a) but also of the driving force for water movement. The Js:k(s-a) ratio was therefore used to estimate axial tension gradients and the radial gradients at a stem height of 1 m were calculated from the differences in axial tension gradients at each depth. Tracheid lumen diameter and tracheid length were used to predict differences in k(s-a) and its divergence from the theoretical k(s-a) determined by the Hagen Poisseuille equation. The ratio of k(s-a) (determined in the laboratory) to Js (measured in the field) varied with depth in the sapwood, resulting in non uniform axial and radial tension gradients from inner to outer sapwood. Transpiration-induced axial tension gradients were in the range of 0.006-0.01 MPa m(-1) excluding the gravitational tension gradient. At a stem height of 1 m, radial tension gradients were in the range of 0.15-0.25 MPa m(-1) and were lower in the middle sapwood than in the inner or outer sapwood. Axial tension gradients were 44-50% higher in the outer sapwood than in the inner sapwood. At a stem height of 1 m, radial Js, calculated on the basis of radial tension gradients and measured radial specific conductivity (k(s-r)), was about two orders of magnitude smaller than axial Js. Our findings indicate that large radial tension gradients occur in the sapwood and clarify the role played by xylem k(s-a) and k(s-r) in determining in situ partitioning of Js in the axial and radial directions. PMID- 16356901 TI - Simulation of effects of wood microstructure on water transport. AB - A tracheid-level model was used to quantify the effects of differences in wood microstructure between coastal and interior Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco var. menziesii and var. glauca) wood on larger scale properties like hydraulic conductivity. The model showed that tracheid length, the ease of flow through a bordered pit and effective tracheid diameter can all limit maximum hydraulic conductivity. Among the model parameters tested, increasing bordered pit conductivity and tracheid length resulted in the greatest increase in maximum conductivity in both the inland and coastal ecotypes. A sensitivity analysis of the uncertainty between parameters governing flow through the bordered pit and air-seeding potential showed that, although decreased pit flow resistance increased maximum hydraulic conductivity, increased cavitation led to lower conductivity over time. The benefits of increasing the number of bordered pits depended on the intensity of the meteorological driving function: in drier environmental conditions, wood with fewer pits was more conductive over time than wood with more pits. Switching the bordered pit characteristics between coastal and interior wood indicated that the conductivity time course of coastal and interior wood was primarily governed by differences in the number of bordered pits and not differences in tracheid dimensions. The rate at which tracheids refilled had little effect on the conductivity time course of either coastal or interior wood during the first two summers when the wood was highly saturated, but had a marked influence in subsequent years once the cavitation profile stabilized. Our work highlights the need for more empirical work on bordered pits to determine whether variation in their number and properties is related to changing environmental conditions. In addition, a detailed simulation model of a bordered pit is needed to understand how variation in pit properties affects the relationship between ease of flow through a bordered pit and its potential for facilitating air-seeding. PMID- 16356902 TI - Usefulness of diurnal trunk shrinkage as a water stress indicator in plum trees. AB - We compared seasonal changes in maximum diurnal trunk shrinkage (MDS) with seasonal changes in midday stem water potential (Psi(s)) over three years in plum trees grown in differing drip-irrigated regimes. In well-irrigated trees, day-to day variations in Psi(s) and MDS were related to evaporative demand. Reference equations were obtained to predict MDS and Psi(s) values for well-irrigated trees as functions of environmental conditions. A decrease in plant water status toward the end of the growing season occurred even in the well-irrigated trees, probably reflecting a reduced volume of soil wetted by the drip irrigation system. Thus, for the prediction of Psi(s), different reference equations are required for the fruit-growth and after-harvest phenological periods. A seasonal change in the relationship between MDS and Psi(s) was observed, which compensated for the decrease in plant water status such that well-irrigated trees had similar MDS values during both the fruit-growth and after-harvest periods. The influence of tree size on the relationship between MDS and Psi(s) was also investigated. For tree trunk diameters ranging between 8 and 13 cm, MDS increased 13% for each cm of increase in trunk diameter, as a result of the thicker phloem tissues of the larger trees. This finding may allow extrapolation of Psi(s) predictions based on empirical relationships with MDS to plum trees of different sizes. PMID- 16356903 TI - Influence of soil texture on hydraulic properties and water relations of a dominant warm-desert phreatophyte. AB - We investigated hydraulic constraints on water uptake by velvet mesquite (Prosopis velutina Woot.) at a site with sandy-loam soil and at a site with loamy clay soil in southeastern Arizona, USA. We predicted that trees on sandy-loam soil have less negative xylem and soil water potentials during drought and a lower resistance to xylem cavitation, and reach E(crit) (the maximum steady-state transpiration rate without hydraulic failure) at higher soil water potentials than trees on loamy-clay soil. However, minimum predawn leaf xylem water potentials measured during the height of summer drought were significantly lower at the sandy-loam site (-3.5 +/- 0.1 MPa; all errors are 95% confidence limits) than at the loamy-clay site (-2.9 +/- 0.1 MPa). Minimum midday xylem water potentials also were lower at the sandy-loam site (-4.5 +/- 0.1 MPa) than at the loamy-clay site (-4.0 +/- 0.1 MPa). Despite the differences in leaf water potentials, there were no significant differences in either root or stem xylem embolism, mean cavitation pressure or Psi(95) (xylem water potential causing 95% cavitation) between trees at the two sites. A soil-plant hydraulic model parameterized with the field data predicted that E(crit) approaches zero at a substantially higher bulk soil water potential (Psi(s)) on sandy-loam soil than on loamy-clay soil, because of limiting rhizosphere conductance. The model predicted that transpiration at the sandy-loam site is limited by E(crit) and is tightly coupled to Psi(s) over much of the growing season, suggesting that seasonal transpiration fluxes at the sandy-loam site are strongly linked to intra annual precipitation pulses. Conversely, the model predicted that trees on loamy clay soil operate below E(crit) throughout the growing season, suggesting that fluxes on fine-textured soils are closely coupled to inter-annual changes in precipitation. Information on the combined importance of xylem and rhizosphere constraints to leaf water supply across soil texture gradients provides insight into processes controlling plant water balance and larger scale hydrologic processes. PMID- 16356904 TI - Excitation energy partitioning and quenching during cold acclimation in Scots pine. AB - We studied the influence of two irradiances on cold acclimation and recovery of photosynthesis in Scots pine (Pinus sylvestris L.) seedlings to assess mechanisms for quenching the excess energy captured by the photosynthetic apparatus. A shift in temperature from 20 to 5 degrees C caused a greater decrease in photosynthetic activity, measured by chlorophyll fluorescence and oxygen evolution, in plants exposed to moderate light (350 micromol m(-2) s(-1)) than in shaded plants (50 micromol m(-2) s(-1)). In response to the temperature shift, maximal photochemical efficiency of photosystem II (PSII), measured as the ratio of variable to maximal chlorophyll fluorescence (Fv/Fm) of dark-adapted samples, decreased to 70% in exposed seedlings, whereas shaded seedlings maintained Fv/Fm close to initial values. After a further temperature decrease to -5 degrees C, only 8% of initial Fv/Fm remained in exposed plants, whereas shaded plants retained 40% of initial Fv/Fm. Seven days after transfer from -5 to 20 degrees C, recovery of photochemical efficiency was more complete in the shaded plants than in the exposed plants (87 and 65% of the initial Fv/Fm value, respectively). In response to cold stress, the estimated functional absorption cross section per remaining PSII reaction center increased at both irradiances, but the increase was more pronounced in exposed seedlings. Estimates of energy partitioning in the needles showed a much higher dissipative component in the exposed seedlings at low temperatures, pointing to stronger development of non-photochemical quenching at moderate irradiances. The de-epoxidation state of the xanthophyll cycle pigments increased in exposed seedlings at 5 degrees C, contributing to the quenching capacity, whereas significant de-epoxidation in the shaded plants was observed only when temperatures decreased to -5 degrees C. Thermoluminescence (TL) measurements of PSII revealed that charge recombinations between the second oxidation state of Mn-cluster S2 and the semireduced secondary electron acceptor quinone Q(B)- (S2Q(B)-) were shifted to lower temperatures in cold-acclimated seedlings compared with control seedlings and this effect depended on irradiance. Concomitant with this, cold-acclimated seedlings demonstrated a significant shift in the S2 recombination with primary acceptor Q(A)- (S2Q(A)-) characteristic TL emission peak to higher temperatures, thus narrowing the redox potential gap between S2Q(B)- and S2Q(A)-, which might result in increased probability for non radiative radical pair recombination between the PSII reaction center chlorophyll a (P680+) and Q(A)- (P680+)Q(A)-) (reaction center quenching) in cold-acclimated seedlings. In Scots pine seedlings, mechanisms of quenching excess light energy in winter therefore involve light-dependent regulation of reaction center content and both reaction center-based and antenna-based quenching of excess light energy, enabling them to withstand high excitation pressure under northern winter conditions. PMID- 16356905 TI - Three-dimensional reconstruction of partially 3D-digitized peach tree canopies. AB - A simplified method for building three-dimensional (3D) mock-ups of peach trees is presented. The method combines partial digitizing of tree structure with reconstruction rules for non-digitized organs. Reconstruction was applied at two scales: leaves on current-year shoots (CYS) and shoots on 1-year-old shoots (OYOS). Reconstruction rules make use of allometric relationships, random sampling of shoot attribute distribution and additional hypotheses (e.g., constant internode length). The method was quantitatively assessed for two training systems (tight goblet and wide-double-Y), at a range of spatial scales. For this purpose, light interception properties of reference and reconstructed mock-ups were compared. Mock-up quality depended on scale. Foliage reconstruction on CYS was unsuitable for generating a given CYS. Similarly, CYS reconstruction on OYOS was unsuitable for generating a given OYOS. This is because generic rules derived at the population scale do not consider specific foliage or shoot attributes of a given CYS or OYOS. In contrast, foliage reconstruction on CYS was able to generate OYOS mock-ups having light properties similar to the reference mock-ups. The same held for CYS reconstruction on OYOS for light capture properties at the tree scale. The CYS reconstruction on OYOS was also suitable for deriving OYOS distribution as a function of light interception ability. Reconstruction rules were successfully used to build the vegetation neighborhood of a reference shoot. The proposed method could therefore be used to make 3D tree mock-ups usable for a range of some, but not all, light computations. Because the simplified method allows large time savings, it could be used in virtual experiments requiring large numbers of replicates, such as comparative studies of tree genotypes or training systems. PMID- 16356906 TI - Physiological responses of three deciduous conifers (Metasequoia glyptostroboides, Taxodium distichum and Larix laricina) to continuous light: adaptive implications for the early Tertiary polar summer. AB - Polar regions were covered with extensive forests during the Cretaceous and early Tertiary, and supported trees comparable in size and productivity to those of present-day temperate forests. With a winter of total or near darkness and a summer of continuous, low-angle illumination, these temperate, high-latitude forests were characterized by a light regime without a contemporary counterpart. Although maximum irradiances were much lower than at mid-latitudes, the 24-h photoperiod provided similar integrated light flux. Taxodium, Larix and Metasequoia, three genera of deciduous conifers that occurred in paleoarctic wet forests, have extant, closely related descendents. However, the contemporary relative abundance of these genera differs greatly from that in the paleoarctic. To provide insight into attributes that favor competitive success in a continuous light environment, we subjected saplings of these genera to a natural photoperiod or a 24-h photoperiod and measured gas exchange, chlorophyll fluorescence, non structural carbohydrate concentrations, biomass production and carbon allocation. Exposure to continuous light significantly decreased photosynthetic capacity and quantum efficiency of photosystem II in Taxodium and Larix, but had minimal influence in Metasequoia. In midsummer, foliar starch concentration substantially increased in both Taxodium and Larix saplings grown in continuous light, which may have contributed to end-product down-regulation of photosynthetic capacity. In contrast, Metasequoia allocated photosynthate to continuous production of new foliar biomass. This difference in carbon allocation may have provided Metasequoia with a two fold advantage in the paleoarctic by minimizing depression of photosynthetic capacity and increasing photosynthetic surface. PMID- 16356907 TI - Within-tree transcriptome profiling in wood-forming tissues of a fast-growing Eucalyptus tree. AB - Despite the availability of high-throughput transcript profiling technology, little is known about tissue-specific gene expression patterns in the wood forming tissues of Eucalyptus plantation tree species. We used cDNA-amplified fragment length polymorphism (AFLP) analysis in combination with infrared fragment detection and semi-automated band quantification to profile gene expression in a 6-year-old, fast- growing Eucalyptus tree. The expression profiles of 6385 transcript-derived fragments (TDFs) were analyzed across four major woody tissues (mature xylem, immature xylem, phloem and cork) collected from two stem positions, to provide a global view of transcript abundance and variability in the Eucalyptus stem. About 21% of the TDFs were differentially expressed and could be grouped into clusters representing co- expressed genes. A total of 71 TDFs representing different gene clusters were isolated and characterized. These included genes implicated in cell fate, signal transduction and cell wall biosynthesis, processes closely associated with xylogenesis. Analysis of the expression levels of selected TDFs by quantitative RT-PCR corroborated the TDF quantification and confirmed that cDNA-AFLP analysis is a highly efficient and accurate tool for transcript profiling and gene discovery in wood-forming tissues of tree species. PMID- 16356908 TI - Hyperhydricity in apple: ultrastructural and physiological aspects. AB - We studied the effects of hyperhydricity on subcellular ultrastructure and physiology of leaves during in vitro regeneration of apple plants. Morphological, anatomical and ultrastructural differences between healthy leaf tissues obtained from greenhouse-grown plants and healthy and hyperhydric leaves obtained from shoots raised from nodal shoot explants in a bioreactor were investigated by electron microscopy and confocal laser scanning microscopy. Compared with healthy leaves, hyperhydric leaves showed abnormal, often discontinuous development of the epidermis and cuticle. Stomata were malformed. The leaf lamina appeared thickened and was characterized by poor differentiation between the palisade and spongy mesophyll tissue. Hyperhydric leaves had a significantly lower chloroplast number per cell and chloroplasts showed reduced thylakoid stacking compared with healthy leaves. Hyperhydricity resulted in a general decrease in concentrations of reduced and oxidized pyridine nucleotides, reflecting a reduction in metabolic activity. The activities of antioxidant enzymes, such as superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were higher in hyperhydric leaves than in healthy leaves, indicating that hyperhydricity was associated with oxidative stress. Chlorophyll fluorescence measurements provided evidence of oxidative damage to the photosynthetic machinery in hyperhydric leaves: photochemical efficiency of photosystem II, effective quantum efficiency and photochemical quenching were all lower in hyperhydric leaves compared with healthy leaves. PMID- 16356909 TI - Interactive effects of shade and irrigation on the performance of seedlings of three Mediterranean Quercus species. AB - Shade and irrigation are frequently used to increase the success of Mediterranean Quercus spp. plantations. However, there is controversy about the combined effects of these treatments on plant performance. We assessed the effects of two irradiances (full sunlight and moderate shade) and two summer watering regimes (high (daily) and low (alternate days)) on leaf and whole-plant traits of 1-year old seedlings of Quercus coccifera, Q. ilex subsp. ballota and Q. faginea grown outdoors for 8.5 months. Leaf traits included measures of morphology, nitrogen concentration, gas exchange and photochemical efficiency, and measures of whole plant traits included biomass allocation patterns, growth phenology, across summer leaf area change and relative growth rate (RGR). Moderate shade reduced leaf mass per area, increased photochemical efficiency, maximum carbon assimilation rate (Amax) and allocation to leaves, and prolonged the growing period in one or more of the species. Daily watering in summer increased Amax of Q. ilex and prolonged the growing period of Q. ilex and Q. faginea. Both treatments tended to increase RGR. The effect of shade was greater in the low watering regime than in the high-watering regime for two of the 15 studied traits, with treatment effects being independent for the remaining 13 traits. Leaf nitrogen and the ability to maintain leaf area after the arid period, rather than biomass allocation traits, explained the variation in seedling RGR. Trait responsiveness to the treatments was low and similar among species and between study scales, being unexpectedly low in Q. faginea leaves. This may be because selective pressures on leaf plasticity act differently in deciduous and evergreen species. We conclude that moderate shade and daily summer watering enhance the performance of Mediterranean Quercus seedlings through species-specific mechanisms. PMID- 16356910 TI - DESPOT, a process-based tree growth model that allocates carbon to maximize carbon gain. AB - We present a new model of tree growth, DESPOT (Deducing Emergent Structure and Physiology Of Trees), in which carbon (C) allocation is adjusted in each time step to maximize whole-tree net C gain in the next time step. Carbon gain, respiration and the acquisition and transport of substitutable photosynthetic resources (nitrogen, water and light) are modeled on a process basis. The current form of DESPOT simulates a uniform, monospecific, self-thinning stand. This paper describes DESPOT and its general behavior in comparison to published data, and presents an evaluation of the sensitivity of its qualitative predictions by Monte Carlo parameter sensitivity analysis. DESPOT predicts determinate height growth and steady stand-level net primary productivity (NPP), but slow declines in aboveground NPP and leaf area index. Monte Carlo analysis, wherein the model was run repeatedly with randomly different parameter sets, revealed that many parameter sets do not lead to sustainable NPP. Of those that do lead to sustainable growth, the ratios at maturity of net to gross primary productivity and of leaf area to sapwood area are highly conserved. PMID- 16356911 TI - How should leaf area, sapwood area and stomatal conductance vary with tree height to maximize growth? AB - Conventional wisdom holds that the ratio of leaf area to sapwood area (L/S) should decline during height (H) growth to maintain hydraulic homeostasis and prevent stomatal conductance (g(s)) from declining. We contend that L/S should increase with H based on a numerical simulation, a mathematical analysis and a conceptual argument: (1) numerical simulation--a tree growth model, DESPOT (Deducing Emergent Structure and Physiology Of Trees), in which carbon (C) allocation is regulated to maximize C gain, predicts L/S should increase during most of H growth; (2) mathematical analysis--the formal criterion for optimal C allocation, applied to a simplified analytical model of whole tree carbon-water balance, predicts L/S should increase with H if leaf-level gas exchange parameters including g(s) are conserved; and (3) conceptual argument- photosynthesis is limited by several substitutable resources (chiefly nitrogen (N), water and light) and H growth increases the C cost of water transport but not necessarily of N and light capture, so if the goal is to maximize C gain or growth, allocation should shift in favor of increasing photosynthetic capacity and irradiance, rather than sustaining g(s). Although many data are consistent with the prediction that L/S should decline with H, many others are not, and we discuss possible reasons for these discrepancies. PMID- 16356912 TI - Anatomical-based defense responses of Scots pine (Pinus sylvestris) stems to two fungal pathogens. AB - We investigated the cellular responses of stem tissues of mature Scots pine (Pinus sylvestris L.) trees to inoculations with two fungal pathogens. The bark beetle vectored fungus, Leptographium wingfieldii Morelet, induced longer lesions in the bark, stronger swelling of polyphenolic parenchyma cells, more polyphenol accumulation and increased ray parenchyma activity compared with the root rot fungus, Heterobasidion annosum (Fr.) Bref., or mechanical wounding. Axial resin ducts in the xylem are a general feature of the preformed defenses of Scots pine, but there was no clear induction of additional traumatic axial resin ducts in response to wounding or fungal infection. The anatomical responses of Scots pine to pathogen infection were localized to the infection site and were attenuated away from bark lesions. The responses observed in Scots pine were compared with published studies on Norway spruce (Picea abies (L.) Karst.) for which anatomically based defense responses have been well characterized. PMID- 16356913 TI - Changes in host chitinase isoforms in relation to wounding and colonization by Heterobasidion annosum: early and strong defense response in 33-year-old resistant Norway spruce clone. AB - We studied the defense reactions of 33-year-old susceptible and resistant clones of Norway spruce (Picea abies (L.) Karst.) to the major root-rot fungus Heterobasidion annosum (Fr.) Bref. and determined if tissue cultures can be used as a model system for studying defense responses of mature trees at the molecular level. Quantitative PCR analysis of genomic DNA obtained from samples taken at different times along the lesion length in living bark indicated that the fungus was present in higher amounts and extended further into the host tissue in the susceptible clone than in the resistant clone. In protein extracts from the same lesion samples, there were differences in temporal and spatial changes in host chitinase isoform profiles between the resistant and susceptible clones. Host chitinase isoforms with pI values approximately 4.8, 4.4 and 3.7 increased more during the first 7 days after wounding and inoculation and extended further along the lesion length in the resistant clone than in the susceptible clone. These results suggest that the time from wounding and infection to induction of defense related expression is shorter in the resistant clone indicating a more efficient host defense response than in the susceptible clone. Tissue cultures from the same clones were not resistant to H. annosum and showed no difference in the timing of the increase in chitinase isoforms in response to the pathogen. However, tissue cultures from both clones showed an increase in chitinase isoforms within 6 to 24 h past inoculation, indicating that increased chitinase expression in response to the pathogen is part of a general defense response common to both mature clones and tissue cultures. PMID- 16356914 TI - Osmotic regulation in leaves and roots of olive trees during a water deficit and rewatering. AB - We evaluated the osmotic adjustment capacity of leaves and roots of young olive (Olea europaea L.) trees during a period of water deficit and subsequent rewatering. The trials were carried out in Basilicata (40 degrees 24' N, 16 degrees 48' E) on 2-year-old self-rooted olive plants (cv. 'Coratina'). Plants were subjected to one of four drought treatments. After 13 days of drought, plants reached mean predawn leaf water potentials of -0.45 +/- 0.015 MPa (control), -1.65 +/- 0.021 (low stress), -3.25 +/- 0.035 (medium stress) and 5.35 +/- 0.027 MPa (high stress). Total osmotic adjustment increased with increasing severity of drought stress. Trees in the high stress treatment showed total osmotic adjustments ranging between 2.4 MPa at 0500 h and 3.8 MPa at 1800 h on the last day of the drought period. Osmotic adjustment allowed the leaves to reach leaf water potentials of about -7.0 MPa. Active osmotic adjustment at predawn decreased during the rewatering period in both leaves and roots. Stomatal conductance and net photosynthetic rate declined with increasing drought stress. Osmotic adjustment in olive trees was associated with active and passive osmotic regulation of drought tolerance, providing an important mechanism for avoiding water loss. PMID- 16356915 TI - Light quality treatments enhance somatic seedling production in three southern pine species. AB - Embryogenic cultures of loblolly pine (Pinus taeda L.), slash pine (Pinus elliottii Engelm.), longleaf pine (Pinus palustris Mill.) and slash pine x longleaf pine hybrids were initiated from immature seeds on an initiation medium containing 13.57 microM 2,4-dichlorophenoxyacetic acid and 2.22 microM benzylaminopurine. Embryogenic cultures proliferated and somatic embryos developed, matured and germinated following a modified protocol and media originally developed for radiata pine (Pinus radiata D. Don.) somatic seedling production. A discrete, light-sensitive pre-germination stage and a later germination (radicle emergence) stage were identified by the differential response of somatic embryos to light of different wavelengths. Different light quality treatments were applied during the pre-germination and germination steps, using cool white fluorescent bulbs or light-emitting diodes (LEDs), or both. In general, red wavelengths provided by LEDs during these steps resulted in higher frequencies of somatic embryo germination (up to 64%) and conversion (up to 50%), longer tap roots and more first-order lateral roots than the standard cool white fluorescent treatments or treatment with blue wavelengths from LEDs. In addition, exposure to red light allowed germination of somatic embryos of some clones that failed to produce germinants under fluorescent light. Germination and conversion were further enhanced by sequential application of cool white fluorescent light and red light, resulting in up to 100% germination and conversion in one experiment. Longleaf pine somatic embryos were especially responsive to the light quality treatments, resulting in the first report of somatic seedling production for this species. PMID- 16356916 TI - Mevalonate kinase activity during different stages of plant regeneration from nodular callus cultures in white pine (Pinus strobus). AB - Mevalonate kinase (MK) catalyzes a step in the isoprenoid biosynthetic pathway, which leads to a huge number of compounds that play important roles in plant growth and development. Here, we report on changes in MK activity in white pine (Pinus strobus L.) during plant regeneration by adventitious shoot organogenesis from cotyledons of mature embryos, including nodular callus induction, shoot formation and rooting. Nodular calli were induced from Pinus strobus (PS) embryos by culture in nodular callus induction medium in a 0-, 8- or 16-h photoperiod. Mevalonate kinase activity peaked in nodular calli after three weeks of culture on nodular callus induction medium in a 16-h photoperiod, whereas frequency of nodular callus formation peaked after 4 weeks of culture on nodular callus induction medium in darkness. During adventitious shoot formation, MK activity peaked in shoots derived from dark-grown nodular calli after 3 weeks on bud formation medium, and frequency of shoot formation was highest in dark-grown nodular calli cultured on bud formation medium for 4 weeks. During rooting, MK activity peaked 2 weeks after transfer of adventitious shoots to rooting medium and rooting frequency was highest in adventitious shoots after 3 weeks on rooting medium. Although during nodular callus induction in darkness MK activity was inversely related to frequency of nodular callus formation, MK activity was highly correlated with frequency of shoot formation and with rooting frequency. The observed increase in MK activity preceding rooting suggests that MK could serve as a marker for rooting of white pine shoots in vitro. PMID- 16356917 TI - Lignification in cell cultures of Pinus radiata: activities of enzymes and lignin topochemistry. AB - Enzymatic and topochemical aspects of lignification were studied in a Pinus radiata D. Don cell culture system that was induced to differentiate tracheary elements and sclereids with lignified secondary cell walls. The activities of the lignin-related enzymes phenylalanine ammonia lyase (PAL; EC 4.3.1.5) and cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) increased concomitantly with cell differentiation, indicating that the increase in enzyme activity was related to lignification of the cell walls and was not induced by stress. This result also indicates that PAL and CAD are suitable markers for tracheary element differentiation in coniferous gymnosperms. To further characterize lignification in this cell culture system, cellular UV-microspectrophotometry and thioacidolysis were employed. Typical UV-absorption spectra of lignin were obtained from the secondary cell walls of the tracheary elements and sclereids and from the compound middle lamella connecting differentiated cells, and the presence of lignin was confirmed by thioacidolysis. Certain aspects of lignin topochemistry in the cell walls of the tracheary elements were similar to cell walls of P. radiata wood, such as the high lignin concentration in the compound middle lamella connecting adjacent cells and the lower lignin concentration in the secondary cell walls. Therefore, the P. radiata cell culture system appears to be well suited to study the formation of lignified secondary cell walls in coniferous gymnosperms. PMID- 16356918 TI - Stomatal characteristics of riparian poplar species in a semi-arid environment. AB - Several native poplar species meet at the margins of their natural distributions in southern Alberta, Canada. In this semi-arid area, poplars are obligate riparian species but they occupy several intergrading ecoregions. Populus deltoides Bartr. ex Marsh predominates in the warmest and driest eastern prairie ecoregions; P. balsamifera L. occupies the cooler and wetter western parkland and montane ecoregions; and P. angustifolia James and hybrids between the species occur in the intermediate grassland ecoregions. We investigated stomatal characteristics of these poplars in 51 genotypes collected across the range of ecoregions and grown in a semi-arid common garden. Stomatal length differed among genotypes within species but did not differ among species, ranging from 19 to 22 microm. Total stomatal densities (adaxial plus abaxial) differed among genotypes within species but were similar among species (290-420 stomata mm(-2)). Single surface stomatal densities differed among species and consequently, the ratio of adaxial:abaxial stomatal density also differed, ranging from 0.94 for P. deltoides to 0.27 for P. balsamifera, with intermediate stomatal density ratios in P. angustifolia and hybrids. In a subsequent study of a subset of the same genotypes, stomatal density was correlated with stomatal conductance (r2 = 0.75) and the conductance ratios differed among species in the same manner as the stomatal density ratios. We conclude that: (1) diverse poplar genotypes respond similarly to a semi-arid environment by producing comparatively small and dense stomata; (2) differences in stomatal density underlie differences in stomatal conductance and differences among species in stomatal density ratio or conductance ratio may reflect adaptation to climatic differences among ecoregions; and (3) there is substantial variation in stomatal characteristics within and among species and hybrids in this area that could be useful for the selection and breeding of poplars adapted to different climatic conditions. PMID- 16356919 TI - Comparative water use by the riparian trees Melaleuca argentea and Corymbia bella in the wet-dry tropics of northern Australia. AB - We examined sources of water and daily and seasonal water use patterns in two riparian tree species occupying contrasting niches within riparian zones throughout the wet-dry tropics of northern Australia: Corymbia bella Hill and Johnson is found along the top of the levee banks and Melaleuca argentea W. Fitzg. is restricted to riversides. Patterns of tree water use (sap flow) and leaf water potential were examined in four trees of each species at three locations along the Daly River in the Northern Territory. Predawn leaf water potential was higher than -0.5 MPa throughout the dry season in both species, but was lower at the end of the dry season than at the beginning of the dry season. Contrary to expectations, predawn leaf water potential was lower in M. argentea trees along the river than in C. bella trees along the levees. In contrast, midday leaf water potential was lower in the C. bella trees than in M. argentea trees. There were no seasonal differences in tree water use in either species. Daily water use was lower in M. argentea trees than in C. bella trees. Whole-tree hydraulic conductance, estimated from the slope of the relationship between leaf water potential and sap flow, did not differ between species. Xylem deuterium concentrations indicated that M. argentea trees along the riverbank were principally reliant on river water or shallow groundwater, whereas C. bella trees along the levee were reliant solely on soil water reserves. This study demonstrated strong gradients of tree water use within tropical riparian communities, with implications for estimating riparian water use requirements and for the management of groundwater resources. PMID- 16356920 TI - Variations in sucrose and ABA concentrations are concomitant with heteroblastic leaf shape changes in a rhythmically growing species (Quercus robur). AB - In rhythmically growing woody species such as common oak (Quercus robur L.), stem growth is discontinuous and a bud forms at regular intervals at the shoot apex. These buds are composed of different types of leaves: laminate, aborted lamina and scale. The change in heteroblastic leaf shape from laminate to aborted lamina leaves is regarded as one of the events marking shoot growth arrest. To better understand the determinism of heteroblastic leaf shape change and thus, of rhythmicity, we studied morphogenetic events during the early days of the second flush of growth in oak, as well as changes in sucrose metabolism and abscisic acid (ABA) concentrations in control plants expressing the heteroblastic leaf shape change and in defoliated plants showing no heteroblastic leaf shape change and producing only laminate leaves. In control plants, the leaf shape change was underway on Day 5 of the second flush with the differentiation of the first two aborted lamina leaves. Sucrose concentration in the apices of control plants decreased between Days 3 and 5 during differentiation of the aborted lamina leaves. An inverse pattern was observed in defoliated plants, suggesting that sucrose acts as a signal triggering heteroblastic leaf shape changes. During the same period, acid cell wall invertase activity was high in young stem and laminate leaves of control plants, whereas the activity remained constant and low in the apices. If the laminate leaves were removed, the increase in apical sucrose concentration was proportionally higher than the decrease in apical acid vacuolar invertase activity, suggesting that, in the absence of young leaves, sucrose is imported to the apex. The sucrose concentration in the apex is therefore likely to be affected by trophic competition with the expanding laminate leaves. The decrease in apex sucrose concentration may be one of the mechanisms driving heteroblastic leaf shape change. Differentiation of aborted lamina leaves was followed by a decrease in the organogenic activity of the shoot apical meristem (SAM) between Days 7 and 9. High concentrations of ABA are associated with differentiation of aborted lamina and scale leaves and with low SAM organogenic activity. Shoot apical meristem organogenic activity remained high and ABA concentration in the apex remained low in defoliated plants producing only photosynthetic leaves. These results suggest that (1) ABA is involved in the gradual conversion of embryonic leaves to abnormal leaves, thereby regulating heteroblastic leaf shape changes and (2) changes in ABA concentration influence the intensity of SAM organogenic activity. Heteroblastic development and therefore rhythmic growth could be the result of competition between apices and laminate leaves, with competition first involving sucrose and thereafter ABA. PMID- 16356921 TI - Metabolic responses to water deficit in two Eucalyptus globulus clones with contrasting drought sensitivity. AB - We compared the metabolic responses of leaves and roots of two Eucalyptus globulus Labill. clones differing in drought sensitivity to a slowly imposed water deficit. Responses measured included changes in concentrations of soluble and insoluble sugars, proline, total protein and several antioxidant enzymes. In addition to the general decrease in growth caused by water deficit, we observed a decrease in osmotic potential when drought stress became severe. In both clones, the decrease was greater in roots than in leaves, consistent with the observed increases in concentrations of soluble sugars and proline in these organs. In roots of both clones, glutathione reductase activity increased significantly in response to water deficit, suggesting that this enzyme plays a protective role in roots during drought stress by catalyzing the catabolism of reactive oxygen species. Clone CN5 has stress avoidance mechanisms that account for its lower sensitivity to drought compared with Clone ST51. PMID- 16356922 TI - Physiological basis of seasonal trend in leaf photosynthesis of five evergreen broad-leaved species in a temperate deciduous forest. AB - The physiological basis of photosynthesis during winter was investigated in saplings of five evergreen broad-leaved species (Camellia japonica L., Cleyera japonica Thunb., Photinia glabra (Thunb.) Maxim., Castanopsis cuspidata (Thunb.) Schottky and Quercus glauca Thunb.) co-occurring under deciduous canopy trees in a temperate forest. We focused on temperature dependence of photosynthetic rate and capacity as important physiological parameters that determine light-saturated rates of net photosynthesis at low temperatures during winter. Under controlled temperature conditions, maximum rates of ribulose bisphosphate carboxylation and electron transport (Vcmax) and Jmax, respectively) increased exponentially with increasing leaf temperature. The temperature dependence of photosynthetic rate did not differ among species. In the field, photosynthetic capacity, determined as Vcmax and Jmax at a common temperature of 25 degrees C (Vcmax(25) and Jmax(25)), increased until autumn and then decreased in species-specific patterns. Values of Vcmax(25) and Jmax(25) differed among species during winter. There was a positive correlation of Vcmax(25) with area-based nitrogen concentration among leaves during winter in Camellia and Photinia. Interspecific differences in Vcmax(25) were responsible for interspecific differences in light saturated rates of net photosynthesis during winter. PMID- 16356923 TI - Distortion of quantitative genomic and expression hybridization by Cot-1 DNA: mitigation of this effect. AB - Cross-hybridization of repetitive sequences in genomic and expression arrays is reported to be suppressed with repeat-blocking nucleic acids (C(o)t-1 DNA). Contrary to expectation, we demonstrated that C(o)t-1 also enhanced non-specific hybridization between probes and genomic targets. When added to target DNA, C(o)t 1 enhanced hybridization (2.2- to 3-fold) to genomic probes containing conserved repetitive elements. In addition to repetitive sequences, C(o)t-1 was found to be enriched for linked single copy (sc) sequences. Adventitious association between these sequences and probes distort quantitative measurements of the probes hybridized to desired genomic targets. Quantitative microarray hybridization studies using C(o)t-1 DNA are also susceptible to these effects, especially for probes that map to genomic regions containing conserved repetitive sequences. Hybridization measurements with such probes are less reproducible in the presence of C(o)t-1 than for probes derived from sc regions or regions containing divergent repeat elements, a finding with significant ramifications for genomic and expression microarray studies. We mitigated the requirement for C(o)t-1 either by hybridizing with computationally defined sc probes lacking repeats or by substituting synthetic repetitive elements complementary to sequences in genomic probes. PMID- 16356924 TI - Integrating probe-level expression changes across generations of Affymetrix arrays. AB - There is an urgent need for bioinformatic methods that allow integrative analysis of multiple microarray data sets. While previous studies have mainly concentrated on reproducibility of gene expression levels within or between different platforms, we propose a novel meta-analytic method that takes into account the vast amount of available probe-level information to combine the expression changes across different studies. We first show that the comparability of relative expression changes and the consistency of differentially expressed genes between different Affymetrix array generations can be considerably improved by determining the expression changes at the probe-level and by considering the latest information on probe-level sequence matching instead of the probe annotations provided by the manufacturer. With the improved probe-level expression change estimates, data from different generations of Affymetrix arrays can be combined more effectively. This will allow for the full exploitation of existing results when designing and analyzing new experiments. PMID- 16356925 TI - Structure and energy of a DNA dodecamer under tensile load. AB - In the last decade, methods to study single DNA molecules under tensile load have been developed. These experiments measure the force required to stretch and melt the double helix and provide insights into the structural stability of DNA. However, it is not easy to directly relate the shape of the force curve to the structural changes that occur in the double helix under tensile load. Here, state of-the-art computer simulations of short DNA sequences are preformed to provide an atomistic description of the stretching of the DNA double helix. These calculations show that for extensions larger that approximately 25% the DNA undergoes a structural transformation and a few base pairs are lost from both the terminal and central part of the helix. This locally melted DNA duplex is stable and can be extended up to approximately 50-60% of the equilibrium length at a constant force. It is concluded that melting under tension cannot be modeled as a simple two-state process. Finally, the important role of the cantilever stiffness in determining the shape of the force-extension curve and the most probable rupture force is discussed. PMID- 16356926 TI - Developing a programmed restriction endonuclease for highly specific DNA cleavage. AB - Specific cleavage of large DNA molecules at few sites, necessary for the analysis of genomic DNA or for targeting individual genes in complex genomes, requires endonucleases of extremely high specificity. Restriction endonucleases (REase) that recognize DNA sequences of 4-8 bp are not sufficiently specific for this purpose. In principle, the specificity of REases can be extended by fusion to sequence recognition modules, e.g. specific DNA-binding domains or triple-helix forming oligonucleotides (TFO). We have chosen to extend the specificity of REases using TFOs, given the combinatorial flexibility this fusion offers in addressing a short, yet precisely recognized restriction site next to a defined triple-helix forming site (TFS). We demonstrate here that the single chain variant of PvuII (scPvuII) covalently coupled via the bifunctional cross-linker N (gamma-maleimidobutryloxy) succinimide ester to a TFO (5'-NH2-[CH2](6 or 12) MPMPMPMPMPPPPPPT-3', with M being 5-methyl-2'-deoxycytidine and P being 5-[1 propynyl]-2'-deoxyuridine), cleaves DNA specifically at the recognition site of PvuII (CAGCTG) if located in a distance of approximately one helical turn to a TFS (underlined) complementary to the TFO ('addressed' site: 5' TTTTTTTCTCTCTCTCN(approximately 10)CAGCTG-3'), leaving 'unaddressed' PvuII sites intact. The preference for cleavage of an 'addressed' compared to an 'unaddressed' site is >1000-fold, if the cleavage reaction is initiated by addition of Mg2+ ions after preincubation of scPvuII-TFO and substrate in the absence of Mg2+ ions to allow triple-helix formation before DNA cleavage. Single base pair substitutions in the TFS prevent addressed DNA cleavage by scPvuII-TFO. PMID- 16356928 TI - Physical and functional interactions of the cytomegalovirus US6 glycoprotein with the transporter associated with antigen processing. AB - The endoplasmic reticulum-resident human cytomegalovirus glycoprotein US6 (gpUS6) inhibits peptide translocation by the transporter associated with antigen processing (TAP) to prevent loading of major histocompatibility complex class I molecules and antigen presentation to CD8+ T cells. TAP is formed by two subunits, TAP1 and TAP2, each containing one multispanning transmembrane domain (TMD) and a cytosolic nucleotide binding domain. Here we reported that the blockade of TAP by gpUS6 is species-restricted, i.e. gpUS6 inhibits human TAP but not rat TAP. Co-expression of human and rat subunits of TAP demonstrates independent binding of gpUS6 to human TAP1 and TAP2, whereas gpUS6 does not bind to rat TAP subunits. gpUS6 associates with preformed TAP1/2 heterodimers but not with unassembled TAP subunits. To locate domains of TAP required for gpUS6 binding and function, we took advantage of reciprocal human/rat intrachain TAP chimeras. Each TAP subunit forms two contact sites within its TMD interacting with gpUS6. The dominant gpUS6-binding site on TAP2 maps to an N-terminal loop, whereas inhibition of peptide transport is mediated by a C-terminal loop of the TMD. For TAP1, two gpUS6 binding domains are formed by loops of the C-terminal TMD. The domain required for TAP inactivation is built by a distal loop of the C terminal TMD, indicating a topology of TAP1 comprising 10 endoplasmic reticulum transmembrane segments. By forming multimeric complexes, gpUS6 reaches the distant target domains to arrest peptide transport. The data revealed a nonanalogous multipolar bridging of the TAP TMDs by gpUS6. PMID- 16356927 TI - The autoregulatory translational control element of poly(A)-binding protein mRNA forms a heteromeric ribonucleoprotein complex. AB - Repression of poly(A)-binding protein (PABP) mRNA translation involves the binding of PABP to the adenine-rich autoregulatory sequence (ARS) in the 5' untranslated region of its own mRNA. In this report, we show that the ARS forms a complex in vitro with PABP, and two additional polypeptides of 63 and 105 kDa. The 63 and 105 kDa polypeptides were identified, as IMP1, an ortholog of chicken zip-code binding polypeptide, and UNR, a PABP binding polypeptide, respectively, by mass spectrometry of the ARS RNA affinity purified samples. Using a modified ribonucleoprotein (RNP) immunoprecipitation procedure we further show that indeed, both IMP1 and UNR bind to the ARS containing reporter RNA in vivo. Although both IMP1 and UNR could bind independently to the ARS RNA in vitro, their RNA-binding ability was stimulated by PABP. Mutational analyses of the ARS show that the presence of four of the six oligo(A) regions of the ARS was sufficient to repress translation and the length of the conserved pyrimidine spacers between the oligo(A) sequences was important for ARS function. The ability of mutant ARS RNAs to form the PABP, IMP1 and UNR containing RNP complex correlates well with the translational repressor activity of the ARS. There is also a direct relationship between the length of the poly(A) RNAs and their ability to form a trimeric complex with PABP, and to repress mRNA translation. UV crosslinking studies suggest that the ARS is less efficient than a poly(A) RNA of similar length, to bind to PABP. We show here that the ARS cannot efficiently form a trimeric complex with PABP; therefore, additional interactions with IMP1 and UNR to form a heteromeric RNP complex may be required for maximal repression of PABP mRNA translation under physiological conditions. PMID- 16356929 TI - Hexose-6-phosphate dehydrogenase knock-out mice lack 11 beta-hydroxysteroid dehydrogenase type 1-mediated glucocorticoid generation. AB - The local generation of active glucocorticoid by NADPH-dependent, 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1) oxoreductase activity, has emerged as an important factor in regulating hepatic glucose output and visceral adiposity. We have proposed that this NADPH is generated within the endoplasmic reticulum by the enzyme hexose-6-phosphate dehydrogenase. To address this hypothesis, we generated mice with a targeted inactivation of the H6PD gene. These mice were unable to convert 11-dehydrocorticosterone (11-DHC) to corticosterone but demonstrated increased corticosterone to 11-DHC conversion consistent with lack of 11beta-HSD1 oxoreductase and a concomitant increase in dehydrogenase activity. This increased corticosterone clearance in the knock-out mice resulted in a reduction in circulating corticosterone levels. Our studies define the critical requirement of hexose-6-phosphate dehydrogenase for 11beta HSD1 oxoreductase activity and add a new dimension to the investigation of 11beta HSD1 as a therapeutic target in patients with the metabolic syndrome. PMID- 16356930 TI - The histone acetyltransferase NCOAT contains a zinc finger-like motif involved in substrate recognition. AB - Nuclear cytoplasmic O-GlcNAcase and acetyltransferase (NCOAT) is a bifunctional enzyme with both glycoside hydrolase and alkyltransferase activity. Its O GlcNAcase active site lies in the N terminus of the enzyme and its histone acetyltransferase (HAT) domain lies in the C terminus. Whereas the HAT domain of the enzyme is catalytically and structurally similar to other acetyltransferases across subfamilies, NCOAT has a motif resembling a zinc finger-like domain unique to the MYST family of HATs. Among the MYST family, this zinc finger, or zinc finger-like domain, is responsible for making contacts with the histone tails within nucleosomes for the HAT to catalyze its respective reaction. Here, we show that NCOAT has the ability to directly associate with both an acetylated and unacetylated histone H4 tail in vitro, and a potential zinc finger-like motif found in NCOAT is implicated in this nucleosomal contact, and is necessary for fully efficient enzymatic activity. Subsequent to the catalysis of acetyltransfer to lysine 8 of histone H4 for the enzyme, however, the substrate is released and NCOAT can no longer bind H4 in our assays. Furthermore, this finger domain by itself is sufficient to bind histone H4. PMID- 16356931 TI - Further insight into S-adenosylmethionine-dependent methyltransferases: structural characterization of Hma, an enzyme essential for the biosynthesis of oxygenated mycolic acids in Mycobacterium tuberculosis. AB - Mycolic acids are major and specific components of the cell envelope of Mycobacteria that include Mycobacterium tuberculosis, the causative agent of tuberculosis. Their metabolism is the target of the most efficient antitubercular drug currently used in therapy, and the enzymes that are involved in the production of mycolic acids represent important targets for the development of new drugs effective against multidrug-resistant strains. Among these are the S adenosylmethionine-dependent methyltransferases (SAM-MTs) that catalyze the introduction of key chemical modifications in defined positions of mycolic acids. Some of these subtle structural variations are known to be crucial for both the virulence of the tubercle bacillus and the permeability of the mycobacterial cell envelope. We report here the structural characterization of the enzyme Hma (MmaA4), a SAM-MT that is unique in catalyzing the introduction of a methyl branch together with an adjacent hydroxyl group essential for the formation of both keto- and methoxymycolates in M. tuberculosis. Despite the high propensity of Hma to proteolytic degradation, the enzyme was produced and crystallized, and its three-dimensional structure in the apoform and in complex with S adenosylmethionine was solved to about 2 A. Thestructuresshowtheimportantroleplayedbythemodificationsfound within mycolic acid SAM-MTs, especially thealpha2-alpha3 motif and the chemical environment of the active site. Essential information with respect to cofactor and substrate binding, selectivity and specificity, and about the mechanism of catalytic reaction were derived. PMID- 16356932 TI - Hepatitis C virus glycoproteins mediate low pH-dependent membrane fusion with liposomes. AB - It has been suggested that the hepatitis C virus (HCV) infects host cells through a pH-dependent internalization mechanism, but the steps leading from virus attachment to the fusion of viral and cellular membranes remain uncharacterized. Here we studied the mechanism underlying the HCV fusion process in vitro using liposomes and our recently described HCV pseudoparticles (pp) bearing functional E1E2 envelope glycoproteins. The fusion of HCVpp with liposomes was monitored with fluorescent probes incorporated into either the HCVpp or the liposomes. To validate these assays, pseudoparticles bearing either the hemagglutinin of the influenza virus or the amphotropic glycoprotein of murine leukemia virus were used as models for pH-dependent and pH-independent entry, respectively. The use of assays based either on fusion-induced dequenching of fluorescent probes or on reporter systems, which produce fluorescence when the virus and liposome contents are mixed, allowed us to demonstrate that HCVpp mediated a complete fusion process, leading to the merging of both membrane leaflets and to the mixing of the internal contents of pseudoparticle and liposome. This HCVpp-mediated fusion was dependent on low pH, with a threshold of 6.3 and an optimum at about 5.5. Fusion was temperature-dependent and did not require any protein or receptor at the surface of the target liposomes. Most interestingly, fusion was facilitated by the presence of cholesterol in the target membrane. These findings clearly indicate that HCV infection is mediated by a pH-dependent membrane fusion process. This paves the way for future studies of the mechanisms underlying HCV membrane fusion. PMID- 16356933 TI - Control of MEF2 transcriptional activity by coordinated phosphorylation and sumoylation. AB - A eukaryotic protein is often subject to regulation by multiple modifications like phosphorylation, acetylation, ubiquitination, and sumoylation. How these modifications are coordinated in vivo is an important issue that is poorly understood but is relevant to many biological processes. We recently showed that human MEF2D (myocyte enhancer factor 2D) is sumoylated on Lys-439. Adjacent to the sumoylation motif is Ser-444, which like Lys-439 is highly conserved among MEF2 proteins from diverse species. Here we present [corrected] several lines of evidence to demonstrate that Ser-444 of MEF2D is required for sumoylation of Lys 439. Histone deacetylase 4 (HDAC4) stimulated this modification by acting through Ser-444. In addition, phosphorylation of Ser-444 by Cdk5, a cyclin-dependent kinase known to inhibit MEF2 transcriptional activity, stimulated sumoylation. Opposing the actions of HDAC4 and Cdk5, calcineurin (also known as protein phosphatase 2B) dephosphorylated Ser-444 and inhibited sumoylation of Lys-439. This phosphatase, however, exerted minimal effects on the phosphorylation catalyzed by ERK5, an extracellular signal-regulated kinase known to activate MEF2D. These results identify [corrected] an essential role for Ser-444 in MEF2D sumoylation and reveal [corrected] a novel mechanism by which calcineurin selectively "edits" phosphorylation at different sites, thereby reiterating that interplay between different modifications represents a general mechanism for coordinated regulation of eukaryotic protein functions in vivo. PMID- 16356934 TI - The BTB-kelch protein LZTR-1 is a novel Golgi protein that is degraded upon induction of apoptosis. AB - Members of the BTB-kelch superfamily play important roles during fundamental cellular processes, such as the regulation of cell morphology, migration, and gene expression. The BTB-kelch protein LZTR-1 is deleted in the majority of DiGeorge syndrome patients and is believed to act as a transcriptional regulator. However, functional and expression profiling studies of LZTR-1 have not been performed thus far. Therefore, we examined the subcellular localization and function of LZTR-1 to gain insights into its biological role. Analysis of the primary structure of the protein revealed six N-terminal kelch motifs and two BTB/POZ domains at the C terminus within LZTR-1. Confocal analysis of the subcellular distribution of LZTR-1 using the Golgi markers GM130, Golgin-97, and TGN46 identified a localization of LZTR-1 exclusively on the cytoplasmic surface of the Golgi network that is mediated by its second BTB/POZ domain. In contrast to most other BTB-kelch proteins, LZTR-1 did not co-localize with actin. Treatment with brefeldin A did not lead to redistribution of LZTR-1 to the endoplasmic reticulum but caused its relocalization in dispersed, punctuated structures that were also positive for GM130. These data demonstrate that LZTR-1 is a Golgi matrix-associated protein. Upon induction of apoptosis, LZTR-1 was phosphorylated on tyrosine residues and subsequently degraded; that could be rescued partially by the addition of the caspase inhibitor Z-VAD-fmk and the proteasome inhibitors lactacystin and MG132. Taken together, our experiments identify LZTR-1 as the first BTB-kelch protein that exclusively localizes to the Golgi network, and the binding of LZTR-1 to the Golgi complex is mediated by its second BTB/POZ domain. PMID- 16356935 TI - Crystal structure of NblA from Anabaena sp. PCC 7120, a small protein playing a key role in phycobilisome degradation. AB - Cyanobacterial light-harvesting complexes, the phycobilisomes, are proteolytically degraded when the organisms are starved for combined nitrogen, a process referred to as chlorosis or bleaching. Gene nblA, present in all phycobilisome-containing organisms, encodes a protein of about 7 kDa that plays a key role in phycobilisome degradation. The mode of action of NblA in this degradation process is poorly understood. Here we presented the 1.8-A crystal structure of NblA from Anabaena sp. PCC 7120. In the crystal, NblA is present as a four-helix bundle formed by dimers, the basic structural units. By using pull down assays with immobilized NblA and peptide scanning, we showed that NblA specifically binds to the alpha-subunits of phycocyanin and phycoerythrocyanin, the main building blocks of the phycobilisome rod structure. By site-directed mutagenesis, we identified amino acid residues in NblA that are involved in phycobilisome binding. The results provided evidence that NblA is directly involved in phycobilisome degradation, and the results allowed us to present a model that gives insight into the interaction of this small protein with the phycobilisomes. PMID- 16356936 TI - Nucleotide sequence and DNA secondary structure, as well as replication protein A, modulate the single-stranded abasic endonuclease activity of APE1. AB - A major role of the multifunctional human Ape1 protein is to incise at apurinic/apyrimidinic (AP) sites in DNA via site-specific endonuclease activity. This nuclease function has been well characterized on double-stranded (ds) DNA substrates, where the complementary strand provides a template for subsequent base excision repair events. Recently, Ape1 was found to incise efficiently at AP sites positioned within the single-stranded (ss) regions of various biologically relevant DNA configurations. The studies within indicated that the ss endonuclease activity of Ape1 is poorly active on ss AP site-containing polyadenine or polythymine oligonucleotides, suggesting a requirement for some form of DNA secondary structure for efficient cleavage. Computational, footprinting, and biochemical analyses indicated that the nature of the secondary structure and the proximity of the AP site influence Ape1 incision efficiency significantly. Replication protein A (RPA), the major ssDNA-binding protein in mammalian cells, was found to bind ss AP-DNA with similar affinity as unmodified ssDNA and ds AP-DNA with lower affinity. Consistent with their known relative DNA binding affinities, RPA blocks/inhibits the ss, but not ds, AP endonuclease function of Ape1. Moreover, RPA inactivates Ape1 incision activity at an AP site within the ss region of a fork duplex, but not a transcription-like bubble intermediate. The data herein suggested a model whereby RPA selectively suppresses the nontemplated ss cleavage activity of Ape1 in vivo, particularly at sites of ongoing replication/recombination, by coating the ssDNA. PMID- 16356937 TI - The carboxyl terminus of the alpha-subunit of the amiloride-sensitive epithelial sodium channel binds to F-actin. AB - The activity of the amiloride-sensitive epithelial sodium channel (ENaC) is modulated by F-actin. However, it is unknown if there is a direct interaction between alpha-ENaC and actin. We have investigated the hypothesis that the actin cytoskeleton directly binds to the carboxyl terminus of alpha-ENaC using a combination of confocal microscopy, co-immunoprecipitation, and protein binding studies. Confocal microscopy of Madin-Darby canine kidney cell monolayers stably transfected with wild type, rat isoforms of alpha-, beta-, and gamma-ENaC revealed co-localization of alpha-ENaC with the cortical F-actin cytoskeleton both at the apical membrane and within the subapical cytoplasm. F-actin was found to co-immunoprecipitate with alpha-ENaC from whole cell lysates of this cell line. Gel overlay assays demonstrated that F-actin specifically binds to the carboxyl terminus of alpha-ENaC. A direct interaction between F-actin and the COOH terminus of alpha-ENaC was further corroborated by F-actin co-sedimentation studies. This is the first study to report a direct and specific biochemical interaction between F-actin and ENaC. PMID- 16356939 TI - Development of a short form of the Workstyle measure. AB - BACKGROUND: 'Workstyle', or how a worker behaviourally, cognitively and physiologically responds to increased or stressful work demands, has been proposed to help explain the link between ergonomic and psychosocial factors in work-related upper limb disorder symptoms and disorders (WRULD). AIM: To describe the psychometric properties of a shortened version of the original Workstyle measure. METHODS: Factor analyses of the Workstyle measure items were conducted to reduce the number of total items. Each of the subscales was then further reduced by randomly selecting half of the items within each subscale. Additionally, two subscales from the original survey (Pain/Tension and Numbness/Tingling) were eliminated because they were not used to calculate the original workstyle total score in order to reduce the influence of current symptoms on an individual's total score. RESULTS: The Workstyle Short Form was reduced to 32 items. Cronbach's alpha was 0.89 and the test-retest reliability was r = 0.88, P < 0.01, for the total score. The short form score was significantly correlated with the full workstyle total score, r = 0.98, P < 0.01. Higher total workstyle scores were significantly associated with pain, functional limitations and adverse mental and physical health. CONCLUSION: The Workstyle Short Form demonstrated acceptable psychometric properties. These findings indicate its potential utility in research on WRULD. PMID- 16356938 TI - Acetylation by p300 regulates nuclear localization and function of the transcriptional corepressor CtBP2. AB - CtBP family members, CtBP1 and CtBP2, are unique transcriptional regulators that adapt a metabolic enzyme fold, and their activities are regulated by NAD(H) binding. CtBP1 is both cytoplasmic and nuclear, and its subcellular localization is regulated by sumoylation, phosphorylation, and binding to a PDZ protein. In contrast, we showed that CtBP2 is exclusively nuclear. CtBP1 and CtBP2 are highly similar, but differ at the N-terminal 20 amino acid region. Substitution of the N terminal domain of CtBP1 with the corresponding CtBP2 domain confers a dominant nuclear localization pattern to CtBP1. The N-terminal domain of CtBP2 contains three Lys residues. Our results show that these Lys residues are acetylated by the nuclear acetylase p300. Although all three Lys residues of CtBP2 (Lys-6, Lys 8, and Lys-10) appear to be acetylated, acetylation of Lys-10 is critical for nuclear localization. CtBP2 with a single amino acid substitution at Lys-10 (K10R) is predominantly localized in the cytoplasm. The cytoplasmic localization of the K10R mutant is correlated with enhanced nuclear export that is inhibited by leptomycin B. Furthermore, lack of acetylation at Lys-10 renders CtBP2 to be more efficient in repression of the E-cadherin promoter. Our studies have revealed the important roles of acetylation in regulating subcellular localization and transcriptional activity of CtBP2. PMID- 16356940 TI - Formation and possible roles of nitric oxide in plant roots. AB - Nitric oxide has been reported to act as a signalling molecule in different plant tissues and to participate in a variety of physiological processes. It is produced by different enzymes and sources. The root-specific plasma membrane bound enzymes forming NO from the substrates nitrate and nitrite are of particular interest because roots serve as interfaces between plants and the soil. The co-ordinated activity of the root-specific plasma membrane-bound nitrate reductase (PM-NR) and nitrite:NO reductase (NI-NOR) suggests that NO might also be involved in root signalling and development. The rate of enzymatic production of this NO depends largely on the environmental conditions, mainly the availability of nitrate and oxygen and it is proposed that this NO plays a role during anoxia as an indicator of the external nitrate availability and in regulating symbiotic interactions at the root surface. PMID- 16356941 TI - Mechanisms for nitric oxide synthesis in plants. AB - The discovery that nitric oxide (NO) acts as a signal fundamentally shifted our understanding of free radicals from toxic by-products of oxidative metabolism to key regulators of cellular functions. This discovery has led to intense investigation into the synthesis of NO in both animals and plants. Nitric oxide synthases (NOS) are the primary sources of NO in animals and are complex, highly regulated enzymes that oxidize arginine to NO and citrulline. Plant NO synthesis, however, appears more complex and includes both nitrite and arginine-dependent mechanisms. The components of the arginine pathway have been elusive as no known orthologues of animal NOS exist in plants. An Arabidopsis gene (AtNOS1) has been identified that is needed for NO synthesis in vivo and has biochemical properties similar to animal cNOS, yet it has no sequence similarity to any known animal NOS. An Atnos1 insertion mutant has been useful for genetic studies of NO regulation and for uncovering new roles for NO signalling. The elucidation of plant NO synthesis promises to yield novel mechanisms that may be applicable to animal systems. PMID- 16356942 TI - Are the metabolic components of crassulacean acid metabolism up-regulated in response to an increase in oxidative burden? AB - In the halophytic species Mesembryanthemum crystallinum, crassulacean acid metabolism (CAM) may be induced by a range of abiotic factors including drought, salinity, high light intensity, low temperature, and anoxia. A key biotic consequence of all these environmental changes is the generation of reactive oxygen species in planta that can elicit potentially damaging oxidative reactions and/or act as signals for engaging mechanisms that alleviate oxidative stress. However, induction of CAM per se also has the potential for increasing the oxidative burden via the enhanced internal O2 concentrations that develop behind closed stomata during daytime decarboxylation. The aim of this paper was to test two hypotheses. The first one, that reactive oxygen species are key signals for up-regulating the major genes and proteins required for the operation of CAM as part of an integrated strategy for alleviating oxidative burden, was tested using gaseous ozone to increase the oxidative burden at a cellular level. The second hypothesis, that CAM potentially increases oxidative load, was tested using a CAM deficient mutant of M. crystallinum. The data indicate that ozone, like salinity, elicits an increase in the transcript and protein abundance of myo-inositol o methyl transferase (a key enzyme of cyclitol synthesis), together with phosphoenolpyruvate carboxylase and other 'CAM-related' enzymes. However, ozone, unlike salinity, does not induce functional CAM, implying that the various metabolic components required for CAM respond to different signals. Comparing the activities of different subcellular isoforms of superoxide dismutase in wild-type and CAM-deficient mutants of M. crystallinum suggests that the induction of CAM potentially curtails the oxidative load in planta. PMID- 16356943 TI - Irradiance and phenotype: comparative eco-development of sun and shade leaves in relation to photosynthetic CO2 diffusion. AB - The subject of this paper, sun leaves are thicker and show higher photosynthetic rates than the shade leaves, is approached in two ways. The first seeks to answer the question: why are sun leaves thicker than shade leaves? To do this, CO2 diffusion within a leaf is examined first. Because affinity of Rubisco for CO2 is low, the carboxylation of ribulose 1,5-bisphosphate is competitively inhibited by O2, and the oxygenation of ribulose 1,5-bisphosphate leads to energy-consuming photorespiration, it is essential for C3 plants to maintain the CO2 concentration in the chloroplast as high as possible. Since the internal conductance for CO2 diffusion from the intercellular space to the chloroplast stroma is finite and relatively small, C3 leaves should have sufficient mesophyll surfaces occupied by chloroplasts to secure the area for CO2 dissolution and transport. This explains why sun leaves are thicker. The second approach is mechanistic or 'how-oriented'. Mechanisms are discussed as to how sun leaves become thicker than shade leaves, in particular, the long-distance signal transduction from mature leaves to leaf primordia inducing the periclinal division of the palisade tissue cells. To increase the mesophyll surface area, the leaf can either be thicker or have smaller cells. Issues of cell size are discussed to understand plasticity in leaf thickness. PMID- 16356944 TI - Systemic suppression of cluster-root formation and net P-uptake rates in Grevillea crithmifolia at elevated P supply: a proteacean with resistance for developing symptoms of 'P toxicity'. AB - Grevillea crithmifolia R. Br. is a species of Proteaceae that is resistant to developing P-toxicity symptoms at phosphorus supplies in the root environment that induce P-toxicity symptoms in the closely related Hakea prostrata (Proteaceae). It was discovered previously that development of P-toxicity symptoms in H. prostrata is related to its low capacity to down-regulate net P uptake rates (i.e. its low plasticity). The plasticity of net P-uptake rates and whole-plant growth responses in G. crithmifolia has now been assessed in two separate experiments: (i) a range of P, from 0 to 200 micromol P d-1, was supplied to whole root systems; (ii) using a split-root design, one root half was supplied with 0, 3, 75, or 225 micromol P d-1, while the other root half invariably received 3 micromol P d-1. Fresh mass was significantly greater in G. crithmifolia plants that had received a greater daily P supply during the pretreatments, but symptoms of P toxicity were never observed. Cluster-root growth decreased from about half the total root fresh mass when the leaf [P] was lowest (c. 0.1 mg P g-1 DM) to complete suppression of cluster-root growth when leaf [P] was 1-2 mg P g-1 DM. Split-root studies revealed that cluster-root initiation and growth, and net P-uptake rates by roots were regulated systemically, possibly by shoot P concentration. It is concluded that, in response to higher P supply, G. crithmifolia does not develop symptoms of P toxicity because of (i) greater plasticity of its net P-uptake capacity, and (ii) its greater plasticity for allocating P to growth and P storage in roots. This ecologically important difference in plasticity is most probably related to a slightly higher nutrient availability in the natural habitat of G. crithmifolia when compared with that of H. prostrata. PMID- 16356947 TI - Montezuma's treasure. PMID- 16356949 TI - Reaching the poor. PMID- 16356950 TI - The health crisis in Russia. PMID- 16356945 TI - Partner notification of chlamydia infection in primary care: randomised controlled trial and analysis of resource use. AB - OBJECTIVE: To evaluate the effectiveness of a practice nurse led strategy to improve the notification and treatment of partners of people with chlamydia infection. DESIGN: Randomised controlled trial. SETTING: 27 general practices in the Bristol and Birmingham areas. PARTICIPANTS: 140 men and women with chlamydia (index cases) diagnosed by screening of a home collected urine sample or vulval swab specimen. INTERVENTIONS: Partner notification at the general practice immediately after diagnosis by trained practice nurses, with telephone follow up by a health adviser; or referral to a specialist health adviser at a genitourinary medicine clinic. MAIN OUTCOME MEASURES: Primary outcome was the proportion of index cases with at least one treated sexual partner. Specified secondary outcomes included the number of sexual contacts elicited during a sexual history, positive test result for chlamydia six weeks after treatment, and the cost of each strategy in 2003 sterling prices. RESULTS: 65.3% (47/72) of participants receiving practice nurse led partner notification had at least one partner treated compared with 52.9% (39/68) of those referred to a genitourinary medicine clinic (risk difference 12.4%, 95% confidence interval -1.8% to 26.5%). Of 68 participants referred to the clinic, 21 (31%) did not attend. The costs per index case were 32.55 pounds sterling for the practice nurse led strategy and 32.62 pounds sterling for the specialist referral strategy. CONCLUSION: Practice based partner notification by trained nurses with telephone follow up by health advisers is at least as effective as referral to a specialist health adviser at a genitourinary medicine clinic, and costs the same. Trial registration Clinical trials: NCT00112255. PMID- 16356951 TI - Long term outcome after traumatic brain injury. PMID- 16356953 TI - Health should be seen as a human right, global campaign says. PMID- 16356954 TI - WTO amendment on generic drugs draws mixed reaction. PMID- 16356955 TI - Dutch doctors are given guidance on sedation. PMID- 16356956 TI - Severe liver disease will increase unless injecting drug users are tested and treated. PMID- 16356963 TI - Cannabis has medical benefits but needs further investigation, says report. PMID- 16356965 TI - Aid often helps richer people in poor countries, World Bank says. PMID- 16356969 TI - Prisoner with AIDS is released to die at home. PMID- 16356970 TI - Funds for vaccination over next decade may fall short by 10bn dollars. PMID- 16356972 TI - EU aims to tackle growing problem of obesity. PMID- 16356973 TI - Junk food advertising contributes to young Americans' obesity. PMID- 16356977 TI - Parental concerns about their child's emotions and behaviour and referral to specialist services: general population survey. PMID- 16356978 TI - Time to pregnancy and sex of offspring: cohort study. PMID- 16356979 TI - Family history of breast cancer and cost of life assurance: a test case comparison of current UK industry practice. PMID- 16356980 TI - Severe placental malaria and maternal shortness, thinness, and small skeletal size in rural Congo: cohort study. PMID- 16356981 TI - Deprivation and volunteering by general practices: cross sectional analysis of a national primary care system. PMID- 16356982 TI - Overprescribing antidepressants to children: pharmacoepidemiological study in primary care. PMID- 16356983 TI - Frozen shoulder. PMID- 16356984 TI - Medical generalists: connecting the map and the territory. PMID- 16356985 TI - Using markets to reform health care. PMID- 16356986 TI - Mental health legislation should respect decision making capacity. PMID- 16356987 TI - Test of capacity has little practical benefit. PMID- 16356988 TI - Who benefits from the new act? PMID- 16356989 TI - Rationing joint replacements: trust's decision seems to be based on prejudice or attributing blame... PMID- 16356990 TI - Rationing in response to NHS deficits: rural patients are likely to be affected most. PMID- 16356991 TI - Risk of gastrointestinal effects with COX-2 inhibitors and NSAIDs: study conclusions do not reflect findings for celecoxib. PMID- 16356992 TI - US experience of smoke-free prisons. PMID- 16356993 TI - Commissioning perhaps shouldn't follow the American way. PMID- 16356994 TI - District general hospitals have a future in truly rural areas. PMID- 16356995 TI - Rationing joint replacements: ...and is false economy resulting in overall damage. PMID- 16356996 TI - Risk of gastrointestinal effects with COX-2 inhibitors and NSAIDs: what does evidence from randomised trials show about celecoxib? PMID- 16356997 TI - Risk of gastrointestinal effects with COX-2 inhibitors and NSAIDs: COX-2 inhibitors were thought of as a safe option. PMID- 16356998 TI - Risk of gastrointestinal effects with COX-2 inhibitors and NSAIDs: why were patients at major risk excluded? PMID- 16356999 TI - Risk of gastrointestinal effects with COX-2 inhibitors and NSAIDs: how strong is the evidence? PMID- 16357000 TI - European subsidies and developing countries. PMID- 16357001 TI - Giving steroids before elective caesarean section: authors respond to editorial. PMID- 16357002 TI - Recessive disorders and consanguineous marriage. PMID- 16357003 TI - I can fly light aircraft, therefore I can anaesthetise? PMID- 16357004 TI - EWTD has negative impact on training for surgeons. PMID- 16357005 TI - New TB vaccine granted orphan drug status. PMID- 16357007 TI - Aspergillus fumigatus conidia inhibit tumour necrosis factor- or staurosporine induced apoptosis in epithelial cells. AB - A major innate immune response to inhaled conidia of the opportunistic pathogen Aspergillus fumigatus (Af) is the synthesis of pro-inflammatory cytokines, which include tumour necrosis factor (TNF)-alpha, a known inducer of apoptosis. Modulation of host cell apoptosis has been reported to be one of the mechanisms whereby pathogens overcome host cell defences. Our study was designed to investigate whether or not Af conidia could modulate apoptosis induced by TNF alpha or staurosporine (STS). Exposure of epithelial cells treated by these inducers and exposed to Af conidia decreased the number of apoptotic cells detected by Annexin V staining, analysis of nuclear morphology, terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end-labelling reaction and immunoblotting. Inhibition of apoptosis by Af conidia was seen in cells of the A549 pneumocyte II line, human tracheal epithelial 16HBE and primary human respiratory cells. Inhibition of apoptosis by Af conidia was also observed when apoptosis was induced by co-cultivating A549 cells with activated human alveolar macrophages. Unlike Af conidia, conidia of Cladosporium cladosporioides as well as latex beads or killed Af conidia have no inhibitory effect on TNF alpha or STS-induced apoptosis. For TNF-induced apoptosis, the observed anti apoptotic effect of Af conidia was found to be associated with a significant reduction of caspase-3. PMID- 16357008 TI - Mutation and expression of PDGFRA and KIT in malignant peripheral nerve sheath tumors, and its implications for imatinib sensitivity. AB - Platelet-derived growth factor receptor alpha (PDGFRalpha) and c-Kit are receptor tyrosine kinases. Both are targets of the tyrosine kinase inhibitor imatinib mesylate which is approved for treatment of some cancers. In order to assess the role of PDGFRalpha and c-Kit in malignant peripheral nerve sheath tumours (MPNST) we examined human tumours for structural alterations, protein and ligand expression. We investigated 34 MPNST, 6 corresponding plexiform neurofibromas (pNF) and 1 MPNST cell culture from 31 patients for mutations and polymorphisms in PDGFRA (exon 2-21) and KIT (exon 9, 11, 13, 17). PDGFRA was amplified in seven tumours from six patients and MPNST cell culture S462. KIT was amplified in five tumours from four patients and in the cell culture. Two MPNST carried somatic PDGFRA mutations in exons coding for the extracellular domain. In addition we detected several polymorphisms in PDGFRA. No point mutations or polymorphisms were detected in the four KIT exons analysed. PDGFRalpha expression was present in 21 of 28 MPNST patients (75%) and the MPNST cell culture. Expression analysis of PDGFRalpha ligands in MPNST and neurofibromas revealed that PDGF-A was more widely expressed than PDGF-B. Focal c-Kit expression was detected in 2 of 29 (7%) MPNST patients. Imatinib treatment of MPNST cell culture S462 exerted a growth inhibitory effect and prevented PDGF-AA induced PDGFRalpha phosphorylation. In summary, PDGFRA, PDGF and KIT dysregulation as well as growth inhibition of cell culture S462 by imatinib may suggest that MPNST patients benefit from treatment with imatinib. PMID- 16357009 TI - Embryonic stem cells in predictive cardiotoxicity: laser capture microscopy enables assay development. AB - Embryonic stem (ES) cells offer unprecedented opportunities for in vitro drug discovery and safety assessment of compounds. Cardiomyocytes derived from ES cells enable development of predictive cardiotoxicity models to increase the safety of novel drugs. Heterogeneity of differentiated ES cells limits the development of reliable in vitro models for compound screening. We report an innovative and robust approach to isolate ES-derived cardiomyocytes using laser microdissection and pressure catapulting (LMPC). LMPC cells were readily applied onto 96-well format in vitro pharmacology assays. The expression of developmental and functional cardiac markers, Nkx 2.5, MLC2V, GATA-4, Connexin 43, Connexin 45, Serca-2a, cardiac alpha actin, and phospholamban, among others, was confirmed in LMPC ES-derived cardiomyocytes. Functional assays exhibited cardiac-like response to increased extracellular calcium (5.4 mM extracellular Ca2+) and L-type calcium channel antagonist (1 microM nifedipine). In conclusion, laser microdissection and pressure catapulting is a robust technology to isolate homogeneous ES-derived cell types from heterogeneous populations applicable to assay development. PMID- 16357010 TI - Olivocerebellar modulation of motor cortex ability to generate vibrissal movements in rat. AB - The vibrissal movements known as whisking are generated in a pulsatile, or non continuous, fashion and comprise sequences of brief regularly spaced movements. These rhythmic timing sequences imply the existence of periodically issued motor commands. As inferior olivary (IO) neurones generate periodic synchronous discharges that could provide the underlying timing signal, this possibility was tested by determining whether the olivocerebellar system modulates motor cortex (MCtx)-triggered whisker movements in rats. Trains of current pulses were applied to MCtx, and the resulting whisker movements were recorded using a high speed video camera. The evoked movement patterns demonstrated properties consistent with the existence of an oscillatory motor driving rhythm. In particular, movement amplitude showed a bell-shaped dependence on stimulus frequency, with a peak at 11.5+/-2.3 Hz. Moreover, movement trajectories showed harmonic and subharmonic entrainment patterns within specific stimulus frequency ranges. By contrast, movements evoked by facial nerve stimulation showed no such frequency dependent properties. To test whether the IO was the oscillator in question, IO neuronal properties were modified in vivo by intra-IO picrotoxin injection, which enhances synchronous oscillatory IO activity and reduces its natural frequency. The ensuing changes in the evoked whisker patterns were consistent with these pharmacological effects. Furthermore, in cerebellectomized rats, oscillatory modulation of MCtx-evoked movements was greatly reduced, and intra-IO picrotoxin injections did not affect the evoked movement patterns. Additionally, multielectrode recording of Purkinje cell complex spikes showed a temporal correlation of olivocerebellar activity during MCtx stimulus trains to evoked movement patterns. In sum, the results indicate that MCtx's ability to generate movements is modulated by an oscillatory signal arising in the olivocerebellar system. PMID- 16357011 TI - WNK3, a kinase related to genes mutated in hereditary hypertension with hyperkalaemia, regulates the K+ channel ROMK1 (Kir1.1). AB - The serine-threonine kinase WNK3 modulates Cl- transport into and out of cells through its regulation of SLC12A cation/Cl- cotransporters, implicating it as (one of) the long-sought Cl-/volume-sensitive kinase(s). Integrators in homeostatic systems regulate structurally diverse but functionally coupled elements. For example, the related kinase WNK4 regulates the Na-Cl co-transporter (NCC), paracellular Cl- flux, and the K+ channel ROMK1 (Kir1.1) to maintain renal NaCl and K+ homeostasis; mutations in PRKWNK4, encoding WNK4, cause a Mendelian disease featuring hypertension and hyperkalemia. It is known that WNK3 is expressed in the nephron's distal convoluted tubule (DCT) and stimulates NCC activity. Here, we show that WNK3 is also expressed in cortical and outer medullary collecting duct principal cells. Accordingly, we tested WNK3's effect on the mediators of NaCl and K+ handling in these nephron segments--the epithelial sodium channel (ENaC), paracellular Cl- flux, and ROMK1--using established model systems. WNK3 did not alter paracellular Cl- flux in tetracycline-responsive MDCK II cells, nor affect amiloride-sensitive currents when co-expressed with ENaC in Xenopus laevis oocytes. However, additional co expression studies in oocytes revealed WNK3 inhibited the renal-specific K+ channel ROMK1 activity greater than 5.5-fold (p < .0001) by altering its plasmalemmal surface expression; WNK3 did not affect ROMK1's conductance or open/closed probability. In contrast, WNK3 had no effect on the activity of the cardiac long-QT syndrome K+ channel KCNQ1/KCNE1 when co-expressed in oocytes. Inhibition of ROMK1 is independent of WNK3's catalytic activity and is mediated by WNK3's carboxyl terminus--a mechanism distinct from its known kinase-dependent activation of NCC. A kinase-inactivating point mutation, or a missense mutation homologous to one in WNK4 that causes disease produced a gain-of-function effect, enhancing WNK3's inhibition of ROMK1 greater than 2.5-fold relative to wild type kinase (p < .0001). The magnitude and specificity of WNK3's effects at both NCC and ROMK1, its co-expression with its targets in the distal nephron, and the established in vivo effect of WNK4 at these same targets provide evidence that WNK3's action is physiologically relevant. WNK3 is likely a component of one of the mechanisms that determines the balance between renal NaCl reabsorption and K+ secretion. PMID- 16357012 TI - Mitochondrial long chain fatty acid oxidation, fatty acid translocase/CD36 content and carnitine palmitoyltransferase I activity in human skeletal muscle during aerobic exercise. AB - Mitochondrial fatty acid transport is a rate-limiting step in long chain fatty acid (LCFA) oxidation. In rat skeletal muscle, the transport of LCFA at the level of mitochondria is regulated by carnitine palmitoyltransferase I (CPTI) activity and the content of malonyl-CoA (M-CoA); however, this relationship is not consistently observed in humans. Recently, fatty acid translocase (FAT)/CD36 was identified on mitochondria isolated from rat and human skeletal muscle and found to be involved in LCFA oxidation. The present study investigated the effects of exercise (120 min of cycling at approximately 60% V(O2peak)) on CPTI palmitoyl CoA and M-CoA kinetics, and on the presence and functional significance of FAT/CD36 on skeletal muscle mitochondria. Whole body fat oxidation rates progressively increased during exercise (P < 0.05), and concomitantly M-CoA inhibition of CPTI was progressively attenuated. Compared to rest, 120 min of cycling reduced (P < 0.05) the inhibition of 0.7, 2, 5 and 10 microM M-CoA by 16%, 21%, 30% and 34%, respectively. Whole body fat oxidation and palmitate oxidation rates in isolated mitochondria progressively increased (P < 0.05) during exercise, and were positively correlated (r = 0.78). Mitochondrial FAT/CD36 protein increased by 63% (P < 0.05) during exercise and was significantly (P < 0.05) correlated with mitochondrial palmitate oxidation rates at all time points (r= 0.41). However, the strongest (P < 0.05) correlation was observed following 120 min of cycling (r = 0.63). Importantly, the addition of sulfo-N-succimidyloleate, a specific inhibitor of FAT/CD36, reduced mitochondrial palmitate oxidation to approximately 20%, indicating FAT/CD36 is functionally significant with respect to LCFA oxidation. We hypothesize that exercise-induced increases in fatty acid oxidation occur as a result of an increased ability to transport LCFA into mitochondria. We further suggest that decreased CPTI M-CoA sensitivity and increased mitochondrial FAT/CD36 protein are both important for increasing whole body fatty acid oxidation during prolonged exercise. PMID- 16357013 TI - Length-dependent changes in voluntary activation, maximum voluntary torque and twitch responses after eccentric damage in humans. AB - To assess the contribution of central and peripheral factors to changes in maximum voluntary force and its length dependence after eccentric muscle damage, voluntary and twitch torque were measured across a wide angular range, along with voluntary activation using twitch interpolation. Isometric torque from both maximum voluntary contractions (MVCs) and paired twitches to motor nerve stimulation were measured from 60 to 150 deg elbow flexion in 10 deg increments in eight subjects. Optimal angles were determined by curve fitting. Each subject then performed eccentric contractions until voluntary torque had decreased by approximately 40%. Measurements were repeated at 2 h, 1 day and 8 days post exercise to follow acute and longer-term changes. Before exercise, the optimal angle was in the mid-range (93+/-10 deg; mean+/-s.d.) for MVCs, and at a more extended elbow angle for the twitch (106+/-6 deg, P < 0.05). Voluntary activation was generally high (> 94%) but depended on elbow angle, with activation being approximately 4% lower at the most flexed compared to the most extended angle. Two hours after exercise, MVCs decreased 40%, while twitch torque declined 70%. All subjects showed a shift in optimal angle to longer muscle lengths for MVCs (17+/-16 deg at 2 h, 14+/-7 deg at day 1, P < 0.05). This shift contributed minimally (approximately 3%) to the reduction in torque at 90 deg, as the torque angle relation was relatively flat around the optimum. The twitch showed a smaller shift (approximately 4 deg) to longer lengths which was not statistically significant. Voluntary activation was significantly impaired in the early stages after exercise (2 h and day 1, P < 0.05), particularly at short muscle lengths. By 8 days after exercise, the optimal angle had returned to pre-exercise values, but MVC, twitch torque and voluntary activation had not fully recovered. Eccentric exercise causes a short-term shift in the optimal angle for MVCs and produces a length-dependent impairment in voluntary activation. Therefore, it appears that both central and peripheral factors limit muscle performance following eccentric damage, with limits to voluntary drive being especially important at short lengths. PMID- 16357014 TI - Comparison of contraction and calcium handling between right and left ventricular myocytes from adult mouse heart: a role for repolarization waveform. AB - In the mammalian heart, the right ventricle (RV) has a distinct structural and electrophysiological profile compared to the left ventricle (LV). However, the possibility that myocytes from the RV and LV have different contractile properties has not been established. In this study, sarcomere shortening, [Ca2+]i transients and Ca2+ and K+ currents in unloaded myocytes isolated from the RV, LV epicardium (LVepi) and LV endocardium (LVendo) of adult mice were evaluated. Maximum sarcomere shortening elicited by field stimulation was graded in the order: LVendo > LVepi > RV. Systolic [Ca2+]i was higher in LVendo myocytes than in RV myocytes. Voltage-clamp experiments in which action potential (AP) waveforms from RV and LVendo were used as the command signal, demonstrated that total Ca2+ influx and myocyte shortening were larger in response to the LVendo AP, independent of myocyte subtypes. Evaluation of possible regional differences in myocyte Ca2+ handling was based on: (i) the current-voltage relation of the Ca2+ current; (ii) sarcoplasmic reticulum Ca2+ uptake; and (iii) mRNA expression of important components of the Ca2+ handling system. None of these were significantly different between RV and LVendo. In contrast, the Ca2+-independent K+ current, which modulates AP repolarization, was significantly different between RV, LVepi and LVendo. These results suggest that these differences in K+ currents can alter AP duration and modulate the [Ca2+]i transient and corresponding contraction. In summary, these findings provide an initial description of regional differences in excitation-contraction coupling in the adult mouse heart [corrected] PMID- 16357015 TI - Mechanisms of the prostaglandin F2alpha-induced rise in [Ca2+]i in rat intrapulmonary arteries. AB - The mechanisms by which prostaglandin F(2alpha) (PGF(2alpha)) increases intracellular Ca2+ concentration [Ca2+]i in vascular smooth muscle remain unclear. We examined the role of store-, receptor- and voltage-operated Ca2+ influx pathways in rat intrapulmonary arteries (IPA) loaded with Fura PE-3. Low concentrations (0.01-1 microM) of PGF(2alpha) caused a transient followed by a plateau rise in [Ca2+]i. Both responses became maximal at 0.1 microM PGF(2alpha). At higher concentrations of PGF(2alpha), a further slower rise in [Ca2+]i was superimposed on the plateau. The [Ca2+]i response to 0.1 microM PGF(2alpha) was mimicked by the FP receptor agonist fluprostenol, whilst the effect of 10 microM PGF(2alpha) was mimicked by the TP receptor agonist U-46619. The plateau rise in [Ca2+]i in response to 0.1 microM PGF(2alpha) was insensitive to diltiazem, and was abolished in Ca2+-free physiological salt solution, and by pretreatment with La3+, 2-APB, thapsigargin or U-73122. The rises in [Ca2+]i in response to 10 microM PGF(2alpha) and 0.01 microM U-46619 were partially inhibited by diltiazem. The diltiazem-resistant components of both of these responses were inhibited by 2 APB and La3+ to an extent which was significantly less than that seen for the response to 0.1 microM PGF(2alpha), and were also much less sensitive to U-73122. The U-46619 response was also relatively insensitive to thapsigargin. When Ca2+ was replaced with Sr2+, the sustained increase in the Fura PE-3 signal to 0.1 microM PGF(2alpha) was abolished, whereas 10 microM PGF(2alpha) and 0.05 microM U 46619 still caused substantial increases. These results suggest that low concentrations of PGF(2alpha) act via FP receptors to cause IP3-dependent Ca2+ release and store operated Ca2+ entry (SOCE). U-46619 and 10-100 microM PGF(2alpha) cause a TP receptor-mediated Ca2+ influx involving both L-type Ca2+ channels and a receptor operated pathway, which differs from SOCE in its susceptibility to La3+, 2-APB and thapsigargin, does not require phospholipase C activation, and is Sr2+ permeable. PMID- 16357016 TI - An electrical analysis of slow wave propagation in the guinea-pig gastric antrum. AB - This paper provides an electrical description of the propagation of slow waves and pacemaker potentials in the guinea-pig gastric antrum in anal and circumferential directions. As electrical conduction between laterally adjacent circular muscle bundles is regularly interrupted, anal conduction of pacemaker potentials was assumed to occur via an electrically interconnected chain of myenteric interstitial cells of Cajal (ICC(MY)). ICC(MY) were also connected resistively to serially connected compartments of longitudinal muscle. Circumferential conduction occurred in a circular smooth muscle bundle that was represented as a chain of electrically connected isopotential compartments: each compartment contained a proportion of intramuscular interstitial cells of Cajal (ICC(IM)) that are responsible for the regenerative component of the slow wave. The circular muscle layer, which contains ICC(IM), and the ICC(MY) network incorporated a mechanism, modelled as a two-stage chemical reaction, which produces an intracellular messenger. The first stage of the reaction is proposed to be activated in a voltage-dependent manner as described by Hodgkin and Huxley; the messenger altered the mean rate of discharge of depolarizing unitary potentials as a function of the concentration of messenger according to a conventional dose-effect relationship. A separate membrane conductance, scaled by the product of an independent voltage-sensitive reaction, was included in the ICC(MY) compartments; this was used to describe the primary component of pacemaker potentials and simulated a delay before the activation of this membrane current. The model generates pacemaker potentials and slow waves with propagation velocities similar to those determined in the physiological experiments described in the accompanying paper. PMID- 16357017 TI - Propagation of slow waves in the guinea-pig gastric antrum. AB - Intracellular recordings were made from the circular layer of the intact muscular wall of the guinea-pig gastric antrum in preparations where much of the corpus remained attached. When two electrodes were positioned parallel to and near to the greater curvature, slow waves were first detected at the oral site and subsequently at the anal site: the oro-anal conduction velocity was found to be 2.5 mm s(-1). When one electrode was positioned near the greater curvature and the other at a circumferential location, slow waves were first detected near the greater curvature and subsequently at the circumferential site: the circumferential conduction velocity was 13.9 mm s(-1). When recordings were made from preparations in which the circular muscle layer had been removed, the oro anal and the circumferential conduction velocities were both about 3.5 mm s(-1). When slow waves were recorded from preparations in which much of the myenteric network of antral interstitial cells (ICC(MY)) had been dissected away, slow waves were first detected near the region of intact ICC(MY) and subsequently at a circumferential location: the circumferential conduction velocity of slow waves in regions devoid of ICC(MY) was 14.7 mm s(-1). When the electrical properties of isolated single bundles of circular muscle were determined, their length constants were about 3 mm and their time constant about 230 ms, giving an asymptotic electrotonic propagation velocity of 25 mm s(-1). Oro-anal electrical coupling between adjacent bundles of circular muscle was found to vary widely: some bundles were well connected to neighbouring bundles whereas others were not. Together the observations suggest that the slow oro-anal progression of slow waves results from a slow conduction velocity of pacemaker potentials in the myenteric network of interstitial cells. The rapid circumferential conduction of slow waves results from the electrical properties of the circular muscle layer which allow intramuscular ICC (ICC(IM)) to support the radial spread of slow waves: regions of high resistance between bundles prevent the anally directed spread of slow waves within the circular layer. PMID- 16357019 TI - EGFR mutation in gefitinib-responsive small-cell lung cancer. PMID- 16357020 TI - The advisory process for anticancer drug regulation: a global perspective. AB - PURPOSE: This paper summarizes the role of external advisors in oncology drug development and regulation from a global perspective. DESIGN: Recently, representatives from the United States Food and Drug Administration, European Medicines Agency, the Japanese Pharmaceuticals and Medical Devices Agency, the Australian Therapeutic Goods Administration and Health Canada held a meeting in conjunction with the American Society of Clinical Oncology meeting. The role of external advisors in oncology drug development and regulation in each of these jurisdictions was presented and discussed. RESULTS: All regulatory bodies described have experience with two forms of outside expertise: advice from individual experts and advice from a group of experts assembled as an advisory group. Regulatory jurisdictions use individual experts variably. In some regions, individual experts provide advice based on knowledge and experience during the drug development phase or in the planning phase for the submission of a drug registration package. In other regions, these individuals serve as external evaluators with the primary responsibility for the review of a clinical trials package submitted for drug registration. Advisory boards have been formalized in all jurisdictions discussed. Advisory boards have a role in discussing specific applications as well as broad policy issues. A common theme is a composition of a core panel of experts with augmentation by additional expertise as needed for consideration of specific scientific questions. In all jurisdictions, advisory board recommendations are not binding on the regulatory body. CONCLUSIONS: Global oncology drug development and registration involves the use of experts by regulatory authorities. The types of experts needed, the expert's role and the transparency of the advisory process reflect the individual needs in different regions. PMID- 16357018 TI - Dependence of cross-bridge kinetics on myosin light chain isoforms in rabbit and rat skeletal muscle fibres. AB - Cross-bridge kinetics underlying stretch-induced force transients was studied in fibres with different myosin light chain (MLC) isoforms from skeletal muscles of rabbit and rat. The force transients were induced by stepwise stretches (< 0.3% of fibre length) applied on maximally Ca2+-activated skinned fibres. Fast fibre types IIB, IID (or IIX) and IIA and the slow fibre type I containing the myosin heavy chain isoforms MHC-IIb, MHC-IId (or MHC-IIx), MHC-IIa and MHC-I, respectively, were investigated. The MLC isoform content varied within fibre types. Fast fibre types contained the fast regulatory MLC isoform MLC2f and different proportions of the fast alkali MLC isoforms MLC1f and MLC3f. Type I fibres contained the slow regulatory MLC isoform MLC2s and the slow alkali MLC isoform MLC1s. Slow MLC isoforms were also present in several type IIA fibres. The kinetics of force transients differed by a factor of about 30 between fibre types (order from fastest to slowest kinetics: IIB > IID > IIA >> I). The kinetics of the force transients was not dependent on the relative content of MLC1f and MLC3f. Type IIA fibres containing fast and slow MLC isoforms were about 1.2 times slower than type IIA fibres containing only fast MLC isoforms. We conclude that while the cross-bridge kinetics is mainly determined by the MHC isoforms present, it is affected by fast and slow MLC isoforms but not by the relative content of MLC1f and MLC3f. Thus, the physiological role of fast and slow MLC isoforms in type IIA fibres is a fine-tuning of the cross-bridge kinetics. PMID- 16357021 TI - Clinically meaningful changes in health-related quality of life in patients diagnosed with hepatobiliary carcinoma. AB - BACKGROUND: To test the reliability, sensitivity to change in biomarkers associated with disease progression and response to treatment, and clinical meaningfulness of the Functional Assessment of Cancer Therapy-Hepatobiliary (FACT Hep) in patients with hepatobiliary carcinoma. PATIENTS AND METHODS: One hundred and fifty-eight patients diagnosed with hepatobiliary carcinoma were prospectively studied. Health-related quality of life (HRQL) was assessed at baseline (prior to treatment), 3-month follow-up (n=55) and 6-month follow-up (n=27). RESULTS: The internal consistency of all the scales of the FACT-Hep were adequate at all time points (>0.75). The FACT-Hep was found to be sensitive to changes in clinical indicators (alkaline phosphate, alpha-fetoprotein, hemoglobin and survival) that reflect disease progression and response to treatment. Combined results from distribution-based and cross-sectional anchor-based analyses provide the following minimally important difference (MID) estimates: FACT-General (FACT-G) subscales=2-3; FACT-G=6-7; Hepatobiliary Cancer Subscale=5 6; FACT-Hep=8-9; Trial Outcome Index=7-8; and FACT-Hepatobiliary Symptom Index=2 3 points. CONCLUSIONS: The FACT-Hep is a reliable instrument that is responsive to clinical indicators of disease progression and response to treatment. The MID estimates can aid interpretation of HRQL data and facilitate sample size calculation in clinical trials. PMID- 16357022 TI - RT-PCR determination of maspin and mammaglobin B in peripheral blood of healthy donors and breast cancer patients. AB - BACKGROUND: The aim of the present study was to evaluate the accuracy of two markers, maspin and mammaglobin B, singly or in combination, to detect breast cancer. To define better the potential and limits of the two markers for diagnostic purposes, blood positivity was analyzed in relation to clinical, pathological and biological tumor characteristics. PATIENTS AND METHODS: The markers were determined in peripheral blood (PB) samples from 27 healthy donors and 140 previously untreated patients using nested reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Positivity for maspin in blood samples was observed in 24% of patients with an 89% specificity. For mammaglobin B, positivity was observed in 7% of patients and never in healthy donors. The presence of maspin was correlated with cell proliferation of the primary tumor (P = 0.015), whereas mammaglobin B positivity correlated with pathological stage (P = 0.013). The presence of either marker was significantly related to nodal status. CONCLUSIONS: Our results indicate that the two markers in association could represent a potentially useful non-invasive tool to detect breast cancer. The validation of these markers as indicators of high risk of relapse is ongoing in a series of patients with an adequate follow-up. PMID- 16357023 TI - Neuropsychological function in high-risk breast cancer survivors after stem-cell supported high-dose therapy versus standard-dose chemotherapy: evaluation of long term treatment effects. AB - BACKGROUND: Studies on cognitive functioning in breast cancer patients point out that a subset of women exhibit chemotherapy-related neuropsychological impairment. Thereby, high-dose therapy may elevate the risk of cognitive dysfunctions. The primary purpose of the study was to evaluate the impact of high dose versus standard-dose chemotherapy on the late neuropsychological outcome in randomized assigned high-risk breast cancer survivors. Next to focusing prevalence, function specificity and extent of cognitive impairment, the question as to whether doses-dependent group differences occur was investigated. PATIENTS AND METHODS: Twenty-four high-dose and 23 standard-dose patients 5 years, on average, after treatment underwent a comprehensive neuropsychological assessment. In addition, 29 early-stage breast cancer patients matched for age, education and time since treatment were recruited as a comparison group. RESULTS: Global cognitive impairment was observed in 8% of high-dose versus 13% of standard-dose compared with 3% of early-stage breast cancer patients. Compared with normative data, all patient groups performed worse on one attention subtest measuring the simple reaction time (P < 0.001 in each case). By contrast, no significant between-group differences on the late neuropsychological outcome were found. CONCLUSIONS: Five years after treatment, standard-dose patients were slightly, but not significantly, more impaired in cognitive performance than high-dose patients. PMID- 16357024 TI - Characterization of familial Waldenstrom's macroglobulinemia. AB - BACKGROUND: Familial clustering of B-cell disorders among Waldenstrom's macroglobulinemia (WM) patients has been reported, though the frequency and any differences in disease manifestation for familial patients remain to be defined. PATIENTS AND METHODS: We therefore analyzed clinicopathological data from 257 consecutive and unrelated WM patients. Forty-eight (18.7%) patients had at least one first-degree relative with either WM (n = 13, 5.1%), or another B-cell disorder including non-Hodgkin's lymphoma (n = 9, 3.5%), myeloma (n = 8, 3.1%), chronic lymphocytic leukemia (n = 7, 2.7%), monoclonal gammopathy of unknown significance (n = 5, 1.9%), acute lymphocytic leukemia (n = 3, 1.2%) and Hodgkin's disease (n = 3, 1.2%). Patients with a familial history of WM or a plasma cell disorder (PCD) were diagnosed at a younger age and with greater bone marrow involvement. RESULTS: Deletions in 6q represented the only recurrent structural chromosomal abnormality and were found in 13% of patients, all non familial cases. Interphase FISH analysis demonstrated deletions in 6q21-22.1 in nearly half of patients, irrespective of familial background. CONCLUSIONS: The above results suggest a high degree of clustering for B-cell disorders among first-degree relatives of patients with WM, along with distinct clinical features at presentation based on familial disease cluster patterns. Genomic studies to delineate genetic predisposition to WM are underway. PMID- 16357026 TI - Assessment of radiofrequency/microwave radiation emitted by the antennas of rooftop-mounted mobile phone base stations. AB - Radiofrequency (RF) and microwave (MW) radiation exposures from the antennas of rooftop-mounted mobile telephone base stations have become a serious issue in recent years due to the rapidly evolving technologies in wireless telecommunication systems. In Malaysia, thousands of mobile telephone base stations have been erected all over the country, most of which are mounted on the rooftops. In view of public concerns, measurements of the RF/MW levels emitted by the base stations were carried out in this study. The values were compared with the exposure limits set by several organisations and countries. Measurements were performed at 200 sites around 47 mobile phone base stations. It was found that the RF/MW radiation from these base stations were well below the maximum exposure limits set by various agencies. PMID- 16357025 TI - Radon concentration measurements in the Amasra coal mine, Turkey. AB - In this study, the results of atmospheric radon measurements that were performed for the Amasra underground coal mine in Zonguldak bituminous coal basin (Turkey) are presented. The radon measurements were performed for 40 days between November 2004 and December 2004 using passive nuclear etched track detectors. The radon concentrations vary from a minimum value 49 Bq m(-3) in a site located at +40 m to a maximum value 223 Bq m(-3) in a site located at -100 m. Mean concentration is 117 (Bq m(-3)). This value is well below the action level of 500-1,500 Bq m( 3) recommended by the International Commission on Radiological Protection (ICRP) (1993). The mean effective dose value for workers of this mine of 3.4 microSv per day was obtained. This result shows that protection against radiological hazards would not be necessary for workers of this mine((2)). PMID- 16357027 TI - Paired-organ and other selected absorbed fraction for the Korean Reference Adult Male model. AB - The authors have developed a mathematical model for calculating internal radionuclide dosimetry for the Korean Reference Adult Male, and have also derived paired-organ and other selected inter-organ photon-specific absorbed fractions for this model. Each lung, kidney and adrenal gland was set as a separate source region even though each of them shares an identical physiological function with their complementary half. The thyroid gland was also set as a source region. Specific absorbed fractions (SAFs) were then derived by selecting 10 photon energies from 0.02 to 4.0 MeV inclusive. For this purpose the Monte Carlo methodology was used, and the derived SAF was compared with the resulting value of MIRD Pamphlet No. 5 and ORNL TM-8381, both of which were derived on the basis of the ICRP-23 reference man. The comparison showed that MIRD No. 5 and ORNL TM 8381 resulted in a higher absorbed fraction, but the phantom created on the basis of the Korean reference man led to a higher SAF. The weight of the organs of the phantom, and the size and location of the trunk seem to account for the differences. The energy-dependent differences in the SAFs are considered to be related to the distance between the source and target regions, the composition of the intervening tissues, and the photon energies and mean free paths. Also, as a result of deriving SAFs after setting each separate lung, kidney and adrenal gland as a source region, it was found that, although they are of the same physiological function, each individual organ serves as a source region on its own. Differences were noted in SAFs exerted on the source and target organs in accordance with the location of the organs, that is, whether they were located to the left or right of the source organs. The SAF derived in this study can be used for a more accurate internal radionuclide dosimetry for Koreans and other Orientals whose physiology, lifestyle and dietary habits are similar to those of Koreans. PMID- 16357028 TI - Comparing the level of dexterity offered by latex and nitrile SafeSkin gloves. AB - An increase in the occurrence of latex allergy has been concurrent with the increasing use of latex gloves by laboratory and healthcare workers. In recent years nitrile gloves have been used to replace latex gloves to prevent latex allergy. Nitrile gloves offer a comparable level of protection against chemical and biological agents and are more puncture resistant. However, if manual dexterity is compromised by nitrile gloves to a greater degree than latex then this may increase the risk of sharps injuries. The Purdue pegboard test, which measures both gross and fine finger dexterity, was used to test the dexterity levels of two glove types used at HPA CEPR; Kimberly-Clark SafeSkin nitrile and latex laboratory gloves. There was a statistically significant 8.6% increase in fine finger dexterity provided by latex compared with nitrile SafeSkin laboratory gloves but no difference in gross dexterity between the glove types. There was no significant relationship between glove dexterity and age or gender. The selection of glove size was influenced by the digit length of participants. Moreover, those with longer, thinner fingers appeared to have an advantage when using nitrile SafeSkin gloves. The level of dexterity provided by latex and nitrile SafeSkin gloves for tasks on a gross dexterity level are comparable and health workers will benefit from the non-allergenic properties of nitrile. For tasks requiring fine finger dexterity nitrile SafeSkin gloves may impede dexterity. Despite this, the degree of restriction appears to have a negligible impact on safety in this study when compared with the risk of latex sensitization and subsequent allergy. In addition to glove material, working practices must also take into account glove size, fit, grip and thickness, as these factors can all influence dexterity. PMID- 16357029 TI - RNAshapes: an integrated RNA analysis package based on abstract shapes. AB - We introduce RNAshapes, a new software package that integrates three RNA analysis tools based on the abstract shapes approach: the analysis of shape representatives, the calculation of shape probabilities and the consensus shapes approach. This new package is completely reimplemented in C and outruns the original implementations significantly in runtime and memory requirements. Additionally, we added a number of useful features like suboptimal folding with correct dangling energies, structure graph output, shape matching and a sliding window approach. PMID- 16357030 TI - CMfinder--a covariance model based RNA motif finding algorithm. AB - MOTIVATION: The recent discoveries of large numbers of non-coding RNAs and computational advances in genome-scale RNA search create a need for tools for automatic, high quality identification and characterization of conserved RNA motifs that can be readily used for database search. Previous tools fall short of this goal. RESULTS: CMfinder is a new tool to predict RNA motifs in unaligned sequences. It is an expectation maximization algorithm using covariance models for motif description, featuring novel integration of multiple techniques for effective search of motif space, and a Bayesian framework that blends mutual information-based and folding energy-based approaches to predict structure in a principled way. Extensive tests show that our method works well on datasets with either low or high sequence similarity, is robust to inclusion of lengthy extraneous flanking sequence and/or completely unrelated sequences, and is reasonably fast and scalable. In testing on 19 known ncRNA families, including some difficult cases with poor sequence conservation and large indels, our method demonstrates excellent average per-base-pair accuracy--79% compared with at most 60% for alternative methods. More importantly, the resulting probabilistic model can be directly used for homology search, allowing iterative refinement of structural models based on additional homologs. We have used this approach to obtain highly accurate covariance models of known RNA motifs based on small numbers of related sequences, which identified homologs in deeply-diverged species. PMID- 16357031 TI - eQTL Explorer: integrated mining of combined genetic linkage and expression experiments. AB - The development of computational resources to visualize and explore data from combined genome-wide expression and linkage studies is essential for the development of testable hypotheses. eQTL Explorer stores expression profiles, linkage data and information from external sources in a relational database and enables simultaneous visualization and intuitive interpretation of the combined data via a Java graphical interface. eQTL Explorer provides a new and powerful tool to interrogate these very large and complex datasets. PMID- 16357032 TI - SEAN: SNP prediction and display program utilizing EST sequence clusters. AB - SEAN is an application that predicts single nucleotide polymorphisms (SNPs) using multiple sequence alignments produced from expressed sequence tag (EST) clusters. The algorithm uses rules of sequence identity and SNP abundance to determine the quality of the prediction. A Java viewer is provided to display the EST alignments and predicted SNPs. PMID- 16357033 TI - EDGE: extraction and analysis of differential gene expression. AB - EDGE (Extraction of Differential Gene Expression) is an open source, point-and click software program for the significance analysis of DNA microarray experiments. EDGE can perform both standard and time course differential expression analysis. The functions are based on newly developed statistical theory and methods. This document introduces the EDGE software package. PMID- 16357034 TI - SoDA: implementation of a 3D alignment algorithm for inference of antigen receptor recombinations. AB - MOTIVATION: The antigen receptors of adaptive immunity-T-cell receptors and immunoglobulins-are encoded by genes assembled stochastically from combinatorial libraries of gene segments. Immunoglobulin genes then experience further diversification through hypermutation. Analysis of the somatic genetics of the immune response depends explicitly on inference of the details of the recombinatorial process giving rise to each of the participating antigen receptor genes. We have developed a dynamic programming algorithm to perform this reconstruction and have implemented it as web-accessible software called SoDA (Somatic Diversification Analysis). RESULTS: We tested SoDA against a set of 120 artificial immunoglobulin sequences generated by simulation of recombination and compared the results with two other widely used programs. SoDA inferred the correct gene segments more frequently than the other two programs. We further tested these programs using 30 human immunoglobulin genes from Genbank and here highlight instances where the recombinations inferred by the three programs differ. SoDA appears generally to find more likely recombinations. PMID- 16357035 TI - HideNseek, a post-genome approach to locate transgenes exemplified in Arabidopsis thaliana. AB - SUMMARY: Determination of transgene location is essential for investigating the effects of position on transgene expression levels and facilitates cloning of the resident gene affected by insertion. Currently used PCR-based approaches for determination of transgene location are relatively complicated and often fail when the transgene is duplicated, rearranged or fragmented. HideNseek is a new bioinformatics tool that allows computation of transgene locations, provided that a suitable genomic restriction enzyme digestion profile is available. Since the new approach is not based on the terminal sequences of the transgene insert, it is less sensitive to transgene duplication, rearrangement or fragmentation. HideNseek has been tested experimentally and by in silico simulation. The experimental example provided here shows that this simple approach is feasible, permitting rapid location of transgenes with little bench work. AVAILABILITY: available on request from the authors. SUPPLEMENTARY DATA: HideNseek input and output examples, experimental procedures and figures showing experimental results are provided as supplementary files: Supplementary material 1, 2, 3 and Supplementary figures (Figs 1 and 2), respectively. Supplementary data is available at Bioinformatics online. PMID- 16357036 TI - Ridom TraceEdit: a DNA trace editor and viewer. AB - Ridom TraceEdit is a cross-platform graphical DNA trace viewer and editor. TraceEdit displays the chromatogram files from Applied Biosystems automated sequencers and files in the Staden SCF format. Incorrect base calls can be edited and saved. PMID- 16357037 TI - Effects of elevated CO2 on the capacity for photosynthesis of a single leaf and a whole plant, and on growth in a radish. AB - The atmospheric concentration of CO2 will probably rise to about 700 micromol mol(-1) by the end of this century. The effects of elevated growth CO2 on photosynthesis are still not fully understood. Effects of elevated growth CO2 on the capacity for photosynthesis of a single leaf and a whole plant were investigated with the radish cultivar White Cherish. The plants were grown under ambient ( approximately 400 micromol mol(-1)) or elevated CO2 ( approximately 750 micromol mol(-1)). The rates of net photosynthesis per leaf area with a whole plant and a single leaf of plants of various ages (15-26 d after planting) were measured under ambient and elevated CO2. The rates of photosynthesis were increased by 20-28% by elevated CO2. There was no effect of elevated growth CO2 on the rate of photosynthesis, clearly indicating no downward acclimation of photosynthesis to elevated CO2. Elevated CO2 increased dry weight accumulation by >27%. The effect of elevated CO2 on other growth characteristics will also be shown. PMID- 16357038 TI - Cytokinin receptors are required for normal development of auxin-transporting vascular tissues in the hypocotyl but not in adventitious roots. AB - Plants alter the architecture of their root systems to adapt to the environment by modulating post-embryonic (lateral and adventitious) root formation and growth. To understand better the genetic basis of this regulation, we screened ethylmethane sulfonate-mutagenized lines of Arabidopsis thaliana for adventitious rooting mutants. One mutant showed retardation of the primary root growth, no production of lateral roots and enhanced formation of adventitious roots. Mapping and genetic complementation revealed that this mutant named wooden leg-3 (wol-3) was an allele of ARABIDOPSIS HISTIDINE KINASE 4 (AHK4), a locus known to encode a cytokinin receptor. Although the vascular system of the primary root and hypocotyl in the wol-3 mutant was aborted, that of the adventitious roots was normally developed. In the hypocotyl of the wol-3 mutant, auxin signals accumulated around the aborted vascular system. The application of auxin to primary roots induced lateral root formation in the wol-3 mutant. Transport of radiolabeled auxin from the top of the hypocotyl to the primary root was inhibited in wol-3. Although only a single amino acid alteration had occurred in AHK4, the root morphology in the wol-3 mutant was quite similar to that in the ahk2 ahk3 ahk4 triple mutant, which is a loss-of-function mutant of the three cytokinin receptors. This implies that the functional disturbance of AHK4 affects the function of the other receptors. Our results suggest that cytokinin receptors are necessary for the formation of auxin-transporting vascular tissues in the hypocotyl, but not in adventitious roots. PMID- 16357039 TI - Phylogenomic analysis identifies red algal genes of endosymbiotic origin in the chromalveolates. AB - Endosymbiosis has spread photosynthesis to many branches of the eukaryotic tree; however, the history of photosynthetic organelle (plastid) gain and loss remains controversial. Fortuitously, endosymbiosis may leave a genomic footprint through the transfer of endosymbiont genes to the "host" nucleus (endosymbiotic gene transfer, EGT). EGT can be detected through comparison of host genomes to uncover the history of past plastid acquisitions. Here we focus on a lineage of chlorophyll c-containing algae and protists ("chromalveolates") that are postulated to share a common red algal secondary endosymbiont. This plastid is originally of cyanobacterial origin through primary endosymbiosis and is closely related among the Plantae (i.e., red, green, and glaucophyte algae). To test these ideas, an automated phylogenomics pipeline was used with a novel unigene data set of 5,081 expressed sequence tags (ESTs) from the haptophyte alga Emiliania huxleyi and genome or EST data from other chromalveolates, red algae, plants, animals, fungi, and bacteria. We focused on nuclear-encoded proteins that are targeted to the plastid to express their function because this group of genes is expected to have phylogenies that are relatively easy to interpret. A total of 708 genes were identified in E. huxleyi that had a significant Blast hit to at least one other taxon in our data set. Forty-six of the alignments that were derived from the 708 genes contained at least one other chromalveolate (i.e., besides E. huxleyi), red and/or green algae (or land plants), and one or more cyanobacteria, whereas 15 alignments contained E. huxleyi, one or more other chromalveolates, and only cyanobacteria. Detailed phylogenetic analyses of these data sets turned up 19 cases of EGT that did not contain significant paralogy and had strong bootstrap support at the internal nodes, allowing us to confidently identify the source of the plastid-targeted gene in E. huxleyi. A total of 17 genes originated from the red algal lineage, whereas 2 genes were of green algal origin. Our data demonstrate the existence of multiple red algal genes that are shared among different chromalveolates, suggesting that at least a subset of this group may share a common origin. PMID- 16357040 TI - Rapid evolution and gene-specific patterns of selection for three genes of spermatogenesis in Drosophila. AB - Hybrid males resulting from crosses between closely related species of Drosophila are sterile. The F1 hybrid sterility phenotype is mainly due to defects occurring during late stages of development that relate to sperm individualization, and so genes controlling sperm development may have been subjected to selective diversification between species. It is also possible that genes of spermatogenesis experience selective constraints given their role in a developmental pathway. We analyzed the molecular evolution of three genes playing a role during the sperm developmental pathway in Drosophila at an early (bam), a mid (aly), and a late (dj) stage. The complete coding region of these genes was sequenced in different strains of Drosophila melanogaster and Drosophila simulans. All three genes showed rapid divergence between species, with larger numbers of nonsynonymous to synonymous differences between species than polymorphisms. Although this could be interpreted as evidence for positive selection at all three genes, formal tests of selection do not support such a conclusion. Departures from neutrality were detected only for dj and bam but not aly. The role played by selection is unique and determined by gene-specific characteristics rather than site of expression. In dj, the departure was due to a high proportion of neutral synonymous polymorphisms in D. simulans, and there was evidence of purifying selection maintaining a high lysine amino acid protein content that is characteristic of other DNA-binding proteins. The earliest spermatogenesis gene surveyed, which plays a role in both male and female gametogenesis, was bam, and its significant departure from neutrality was due to an excess of nonsynonymous substitutions between species. Bam is degraded at the end of mitosis, and rapid evolutionary changes among species might be a characteristic shared with other degradable transient proteins. However, the large number of nonsynonymous changes between D. melanogaster and D. simulans and a phylogenetic comparative analysis among species confirms evidence of positive selection driving the evolution of Bam and suggests an yet unknown germ cell line developmental adaptive change between these two species. PMID- 16357041 TI - Energy expenditure and body composition of chronically maintained decerebrate rats in the fed and fasted condition. AB - The contribution of the caudal brainstem to adaptation to starvation was tested using chronically maintained decerebrate (CD) and neurologically intact controls. All rats were gavage fed an amount of diet that maintained weight gain in controls. CD rats were subjected to a two-stage surgery to produce a complete transection of the neuroaxis at the mesodiencephalic juncture. One week later, the rats were housed in an indirect calorimeter, and 24 h energy expenditure was measured for 4 d. One half of each of the CD and control groups was then starved for 48 h. Fed CD rats maintained a lower body temperature (35 C), a similar energy expenditure per unit fat-free mass but an elevated respiratory quotient compared with controls. They gained less weight, had 20% less lean tissue, and had 60% more fat than controls. Circulating leptin, adiponectin, and insulin were elevated, glucose was normal, but testosterone was dramatically reduced. Responses to starvation were similar in CD and controls; they reduced energy expenditure, decreased respiratory quotient, indicating lipid utilization, defended body temperature, mobilized fat, decreased serum leptin and insulin, and regulated plasma glucose. These data clearly demonstrate that the isolated caudal brainstem is sufficient to mediate many aspects of the energetic response to starvation. In intact animals, these responses may be refined by a contribution by more rostral brain areas or by communication between fore- and hind-brain. In the absence of communication from the forebrain, the caudal brainstem is inadequate for maintenance of testosterone levels or lean tissue in fed or fasted animals. PMID- 16357042 TI - Arginine 276 controls the directional preference of AKR1C9 (rat liver 3alpha hydroxysteroid dehydrogenase) in human embryonic kidney 293 cells. AB - Rat liver AKR1C9 is the best-studied 3alpha-hydroxysteroid dehydrogenase (3alphaHSD) of the aldo-keto reductase superfamily. The physiologic function of AKR1C9 is to catalyze the reduction of 5alpha-androstane-17beta-ol-3-one (dihydrotestosterone) to 5alpha-androstane-3alpha,17beta-diol (androstanediol) rather than the reverse reaction, and all of the known AKR1C enzymes with 3alphaHSD activity also preferentially catalyze dihydrotestosterone reduction in intact cells. Because the utilization of pyridine-nucleotide cofactors NAD(P)(H) primarily governs the directional preference of HSD enzymes in intact cells, and because R276 participates in NADP(H) binding, we hypothesized that mutation of R276 would alter directional preference in intact cells. To test this model, we constructed stable lines of human embryonic kidney 293 cells expressing wild-type AKR1C9 and mutations R276M, R276G, and R276E. Mutations R276M and R276G retained reductive preference with slightly reduced magnitude compared with wild-type AKR1C9. NADPH depletion by glucose deprivation minimally altered the equilibrium steroid distribution for wild-type AKR1C9 but further reduced the reductive preference of mutations R276M and R276G. Mutation R276E, in contrast, showed an oxidative preference under all conditions. The intrinsic rates of the reductive and oxidative reactions for all four enzymes were similar at the functional equilibrium states. We conclude the R276 maximizes the reductive preference of AKR1C9 in intact cells and maintains this strong preference despite NADPH depletion; mutation R276E reverses the directional preference. PMID- 16357043 TI - Peroxisome proliferator-activated receptor alpha mediates the effects of high-fat diet on hepatic gene expression. AB - Peroxisome proliferator-activated receptors (PPARs) are transcription factors involved in the regulation of numerous metabolic processes. The PPARalpha isotype is abundant in liver and activated by fasting. However, it is not very clear what other nutritional conditions activate PPARalpha. To examine whether PPARalpha mediates the effects of chronic high-fat feeding, wild-type and PPARalpha null mice were fed a low-fat diet (LFD) or high-fat diet (HFD) for 26 wk. HFD and PPARalpha deletion independently increased liver triglycerides. Furthermore, in wild-type mice HFD was associated with a significant increase in hepatic PPARalpha mRNA and plasma free fatty acids, leading to a PPARalpha-dependent increase in expression of PPARalpha marker genes CYP4A10 and CYP4A14. Microarray analysis revealed that HFD increased hepatic expression of characteristic PPARalpha target genes involved in fatty acid oxidation in a PPARalpha-dependent manner, although to a lesser extent than fasting or Wy14643. Microarray analysis also indicated functional compensation for PPARalpha in PPARalpha null mice. Remarkably, in PPARalpha null mice on HFD, PPARgamma mRNA was 20-fold elevated compared with wild-type mice fed a LFD, reaching expression levels of PPARalpha in normal mice. Adenoviral overexpression of PPARgamma in liver indicated that PPARgamma can up-regulate genes involved in lipo/adipogenesis but also characteristic PPARalpha targets involved in fatty acid oxidation. It is concluded that 1) PPARalpha and PPARalpha-signaling are activated in liver by chronic high-fat feeding; and 2) PPARgamma may compensate for PPARalpha in PPARalpha null mice on HFD. PMID- 16357044 TI - Differential expression and processing of chromogranin A and secretogranin II in relation to the secretory status of endocrine cells. AB - Chromogranin A (CgA) and secretogranin II (SgII) are neuroendocrine secretory proteins that participate in regulation of the secretory pathway and also serve as precursors of biologically active peptides. To investigate whether there is a relationship between the expression, distribution, and processing of CgA and SgII and the degree of secretory activity, we employed two melanotrope subpopulations of the pituitary intermediate lobe that exhibit opposite secretory phenotypes. Thus, although one of the melanotrope subtypes shows high secretory activity, the other exhibits characteristics of a hormone storage phenotype. Our data show that SgII expression levels were higher in secretory melanotropes, whereas CgA expression showed similar rates in both cell subsets. The use of various antibodies revealed the presence of the unprocessed proteins as well as three CgA derived peptides (67, 45, and 30 kDa) and six SgII-derived peptides (81, 66, 55, 37, 32, and 30 kDa) in both subpopulations. However, the smallest molecular forms of both granins predominated in secretory melanotropes, whereas the largest SgII- and CgA-immunoreactive peptides were more abundant in storage melanotropes, which is suggestive of a more extensive processing of granins in the secretory subset. Confocal microscopy studies showed that CgA immunoreactivity was higher in storage cells, but SgII immunoreactivity was higher in secretory melanotropes. Taken together, our results indicate that SgII and CgA are differentially regulated in melanotrope subpopulations. Thus, SgII expression is strongly related to the secretory activity of melanotrope cells, whereas CgA expression may not be related to secretory rate, but, rather, to hormone storage in this endocrine cell type. PMID- 16357045 TI - Mechanisms underlying the diminished sensitivity to prolactin negative feedback during lactation: reduced STAT5 signaling and up-regulation of cytokine-inducible SH2 domain-containing protein (CIS) expression in tuberoinfundibular dopaminergic neurons. AB - Hyperprolactinaemia during lactation is a consequence of the sucking stimulus and in part due to reduced prolactin (PRL) negative feedback. To date, the mechanisms involved in this diminished sensitivity to PRL feedback are unknown but may involve changes in PRL signal transduction within tuberoinfundibular dopaminergic (TIDA) neurons. Therefore, we investigated signal transducers and activators of transcription (STAT) 5 signaling in the TIDA neurons of lactating rats. Dual label confocal immunofluorescence studies were used to determine the intracellular distribution of STAT5 within TIDA neurons in the dorsomedial arcuate nucleus. In lactating rats with pups removed for 16 h, injection of ovine PRL significantly (P < 0.05) increased the STAT5 nuclear/cytoplasmic ratio compared with vehicle-treated mothers. In contrast, ovine PRL injection did not increase the STAT5 nuclear/cytoplasmic ratio in lactating mothers with pups, demonstrating that PRL signal transduction through STAT5 is reduced in TIDA neurons in the presence of pups. To investigate possible mechanisms involved in reduced PRL signaling, we examined the expression of suppressors of cytokine signaling (SOCS) proteins. Northern analysis on whole hypothalamus showed that CIS (cytokine-inducible SH2 domain-containing protein), but not SOCS1 or SOCS3, mRNA expression was significantly (P < 0.01) up-regulated in suckled lactating rats. Semiquantitative RT-PCR on arcuate nucleus micropunches also showed up regulation of CIS transcripts. Immunofluorescence studies demonstrated that CIS is expressed in all TIDA neurons in the dorsomedial arcuate nucleus, and the intensity of CIS staining in these neurons is significantly (P < 0.05) increased in lactating rats with sucking pups. Together, these results support the hypothesis that loss of sensitivity to PRL-negative feedback during lactation is a result of increased CIS expression in TIDA neurons. PMID- 16357046 TI - Somatostatin inhibits oxidative respiration in pancreatic beta-cells. AB - Somatostatin potently inhibits insulin secretion from pancreatic beta-cells. It does so via activation of ATP-sensitive K+-channels (KATP) and G protein regulated inwardly rectifying K+-channels, which act to decrease voltage-gated Ca2+-influx, a process central to exocytosis. Because KATP channels, and indeed insulin secretion, is controlled by glucose oxidation, we investigated whether somatostatin inhibits insulin secretion by direct effects on glucose metabolism. Oxidative metabolism in beta-cells was monitored by measuring changes in the O2 consumption (DeltaO2) of isolated mouse islets and MIN6 cells, a murine-derived beta-cell line. In both models, glucose-stimulated DeltaO2, an effect closely associated with inhibition of KATP channel activity and induction of electrical activity (r > 0.98). At 100 nm, somatostatin abolished glucose-stimulated DeltaO2 in mouse islets (n = 5, P < 0.05) and inhibited it by 80 +/- 28% (n = 17, P < 0.01) in MIN6 cells. Removal of extracellular Ca2+, 5 mm Co2+, or 20 microm nifedipine, conditions that inhibit voltage-gated Ca2+ influx, did not mimic but either blocked or reduced the effect of the peptide on DeltaO2. The nutrient secretagogues, methylpyruvate (10 mm) and alpha-ketoisocaproate (20 mm), also stimulated DeltaO2, but this was unaffected by somatostatin. Somatostatin also reversed glucose-induced hyperpolarization of the mitochondrial membrane potential monitored using rhodamine-123. Application of somatostatin receptor selective agonists demonstrated that the peptide worked through activation of the type 5 somatostatin receptor. In conclusion, somatostatin inhibits glucose metabolism in murine beta-cells by an unidentified Ca2+-dependent mechanism. This represents a new signaling pathway by which somatostatin can inhibit cellular functions regulated by glucose metabolism. PMID- 16357047 TI - The interaction of TRbeta1-N terminus with steroid receptor coactivator-1 (SRC-1) serves a full transcriptional activation function of SRC-1. AB - Steroid receptor coactivator-1 (SRC-1) plays a crucial role in nuclear receptor mediated transcription including thyroid hormone receptor (TR)-dependent gene expression. Interaction of the TR-ligand binding domain and SRC-1 through LXXLL motifs is required for this action. However, potential interactions between the TRbeta1-N terminus (N) and SRC-1 have not been explored and thus are examined in this manuscript. Far-Western studies showed that protein construct containing TRbeta1-N + DNA binding domain (DBD) bound to nuclear receptor binding domain (NBD)-1 (amino acid residue, aa 595-780) of SRC-1 without ligand. Mammalian two hybrid studies showed that NBD-1, as well as SRC-1 (aa 595-1440), bound to TRbeta1-N+DBD in the absence of ligand in CV-1 cells. However, NBD-2 (aa 1237 1440) did not bind to this protein. Glutathione-S-transferase pull-down studies showed that TRbeta1-N (aa 1-105) bound to the broad region of SRC-1-C terminus. Expression vectors encoding a series of truncations and/or point mutations of TRbeta1 were used in transient transfection-based reporter assays in CV-1 cells. N-terminal truncated TRbeta1 (DeltaN-TRbeta1) showed lower activity than that of wild-type in both artificial F2-thyroid hormone response element and native malic enzyme response element. These results suggest that there is the interaction between N terminus of TRbeta1 and SRC-1, which may serve a full activation of SRC 1, together with activation function-2 on TRbeta1-mediated transcription. PMID- 16357048 TI - Is glucocorticoid-induced osteonecrosis after kidney transplantation related to osteoporosis? PMID- 16357049 TI - Renal cortical necrosis after kidney transplantation associated with the prothrombin 20210A mutation. PMID- 16357050 TI - Hyperlactataemia induced by CVVHDF with low lactate bicarbonate-buffered solutions in patients with liver dysfunction. PMID- 16357051 TI - Haemoperitoneum in a pregnant woman with peritoneal dialysis. PMID- 16357052 TI - Estimation of trisodium citrate (Citra-Lock) remaining in central venous catheters after the interdialytic interval. PMID- 16357053 TI - Survival and infection rates of a polyurethane vascular access graft compared to tunnelled dialysis catheters and brachiobasilic arteriovenous fistulas. PMID- 16357054 TI - Comparative analysis of rDNA distribution in chromosomes of various species of Brassicaceae. AB - BACKGROUND AND AIMS: The Brassicaceae family encompasses numerous species of great agronomic importance, belonging to such genera, as Brassica, Raphanus, Sinapis and Armoracia. Many of them are characterized by extensive intraspecific diversity of phenotypes. The present study focuses on the polymorphism of number, appearance and chromosomal localization of ribosomal DNA (rDNA) sites and, when possible, in relation to polyploidy, in 42 accessions of Brassica species and ten accessions of Diplotaxis, Eruca, Raphanus and Sinapis species. METHODS: Chromosomal localization of ribosomal DNA was carried out using dual colour fluorescence in situ hybridization (FISH) with 5S rDNA and 25S rDNA sequences as probes on enzymatically digested root-tip meristematic cells. KEY RESULTS: Loci for 5S and 18S-5.8S-25S rDNA were determined for the first time in six taxa, and previously unreported rDNA constellations were described in an additional 12 accessions. FISH revealed frequent polymorphism in number, appearance and chromosomal localization of both 5S and 25S rDNA sites. This phenomenon was most commonly observed in the A genome of Brassica, where it involves exclusively pericentromeric sites of 5S and 25S rRNA genes. The intraspecific polymorphism was between subspecies/varieties or within a variety or cultivar (i.e. interindividual). CONCLUSIONS: The number of rDNA sites can differ up to 5-fold in species with the same chromosome number. In addition to the eight previously reported chromosomal types with ribosomal genes, three new variant types are described. The extent of polymorphism is genome dependent. Comparing the A, B and C genomes revealed the highest rDNA polymorphism in the A genome. The loci carrying presumably inactive ribosomal RNA genes are particularly prone to polymorphism. It can also be concluded that there is no obvious polyploidization related tendency to reduce the number of ribosomal DNA loci in the allotetraploid species, when compared with their putative diploid progenitors. The observed differences are rather caused by the prevailing polymorphism within the diploids and allotetraploids. This would make it difficult to predict expected numbers of rDNA loci in natural polyploids. PMID- 16357055 TI - Diversity of flowering and fruiting phenology of trees in a tropical deciduous forest in India. AB - BACKGROUND AND AIMS: In the dry tropics, vegetative phenology varies widely with tree characteristics and soil conditions. The present work aims to document the phenological diversity of flowering and fruiting with reference to leafing events in Indian dry-tropical tree species. METHODS: Nine tree species, including one leaf-exchanging and eight deciduous showing varying leafless periods, were studied. Monthly counts of leaves, flowers and fruits were made on 160 tagged twigs on ten individuals of each species for initiation, completion and duration of different phenological events through two annual cycles. KEY RESULTS: Variation in flowering relative to leaf flushing (which occurred just prior to or during a hot, dry summer) revealed five flowering types: summer flowering (on foliated shoots), rainy-season flowering (on foliated shoots following significant rains), autumn flowering (on shoots with mature leaves), winter flowering (on shoots undergoing leaf fall) and dry-season flowering (on leafless shoots). Duration of the fruiting phenophase was shortest (3-4 months) in dry season and winter-flowering species, 6-9 months in rainy-and autumn-flowering species, and maximum (11 months) in summer-flowering species. A wide range of time lag (<1 to >8 months) between the start of vegetative (first-leaf flush) and reproductive (first-visible flower) phases was recorded in deciduous species; this time lag was correlated with the extent of the leafless period. A synthesis of available phenological information on 119 Indian tropical trees showed that summer-flowering species were most abundant (56 % of total species) amongst the five types recognized. CONCLUSIONS: The wide diversity of seasonal flowering and fruiting with linkages to leaf flush time and leafless period reflect the fact that variable reproductive and survival strategies evolved in tree species under a monsoonic bioclimate. Flowering periodicity has evolved as an adaptation to an annual leafless period and the time required for the fruit to develop. The direct relationship between leafless period (inverse of growing period) and time lag between onset of vegetative and reproductive phases reflects the partitioning of resource use for supporting these phases. Predominance of summer flowering coupled with summer leaf flushing seems to be a unique adaptation in trees to survive under a strongly seasonal tropical climate. PMID- 16357056 TI - Localization of the sulfate/anion exchanger in the rat liver. AB - Although the sulfate/anion transporter (sat-1; SLC26A1) was isolated from a rat liver cDNA library by expression cloning, localization of sat-1 within the liver and its contribution to the transport of sulfate and organo sulfates have remained unresolved. In situ hybridization and immunohistochemical studies were undertaken to demonstrate the localization of sat-1 in liver tissue. RT-PCR studies on isolated hepatocytes and liver endothelial and stellate cells in culture were performed to test for the presence of sat-1 in these cells. In sulfate uptake and efflux experiments, the substrate specificity of sat-1 was evaluated. Sat-1 mRNA was found in hepatocytes and endothelial cells. Sat-1 protein was localized in sinusoidal membranes and along the borders of hepatocytes. The canalicular region and bile capillaries were not stained. Sulfate uptake was only slightly affected by sulfamoyl diuretics or organo sulfates. Sulfate efflux from sat-1-expressing oocytes was enhanced in the presence of bicarbonate, indicating sulfate/bicarbonate exchange. Estrone sulfate was not transported by sat-1. Sat-1 may be responsible for the uptake of inorganic sulfate from the blood into hepatocytes to enable sulfation reactions. In hepatocytes and endothelial cells, sat-1 may also supply sulfate for proteoglycan synthesis. PMID- 16357057 TI - Coordinated induction of bile acid detoxification and alternative elimination in mice: role of FXR-regulated organic solute transporter-alpha/beta in the adaptive response to bile acids. AB - The bile acid receptor farnesoid X receptor (FXR) is a key regulator of hepatic defense mechanisms against bile acids. A comprehensive study addressing the role of FXR in the coordinated regulation of adaptive mechanisms including biosynthesis, metabolism, and alternative export together with their functional significance is lacking. We therefore fed FXR knockout (FXR(-/-)) mice with cholic acid (CA) and ursodeoxycholic acid (UDCA). Bile acid synthesis and hydroxylation were assessed by real-time RT-PCR for cytochrome P-450 (Cyp)7a1, Cyp3a11, and Cyp2b10 and mass spectrometry-gas chromatography for determination of bile acid composition. Expression of the export systems multidrug resistance proteins (Mrp)4-6 in the liver and kidney and the recently identified basoalteral bile acid transporter, organic solute transporter (Ost-alpha/Ost-beta), in the liver, kidney, and intestine was also investigated. CA and UDCA repressed Cyp7a1 in FXR(+/+) mice and to lesser extents in FXR(-/-) mice and induced Cyp3a11 and Cyp2b10 independent of FXR. CA and UDCA were hydroxylated in both genotypes. CA induced Ost-alpha/Ost-beta in the liver, kidney, and ileum in FXR(+/+) but not FXR(-/-) mice, whereas UDCA had only minor effects. Mrp4 induction in the liver and kidney correlated with bile acid levels and was observed in UDCA-fed and CA fed FXR(-/-) animals but not in CA-fed FXR(+/+) animals. Mrp5/6 remained unaffected by bile acid treatment. In conclusion, we identified Ost-alpha/Ost beta as a novel FXR target. Absent Ost-alpha/Ost-beta induction in CA-fed FXR(-/ ) animals may contribute to increased liver injury in these animals. The induction of bile acid hydroxylation and Mrp4 was independent of FXR but could not counteract liver toxicity sufficiently. Limited effects of UDCA on Ost alpha/Ost-beta may jeopardize its therapeutic efficacy. PMID- 16357058 TI - Regulation of the mouse organic solute transporter alpha-beta, Ostalpha-Ostbeta, by bile acids. AB - The mechanisms responsible for bile acid regulation of mouse intestinal organic solute transporter alpha-beta (Ostalpha-Ostbeta) expression were investigated. Expression of Ostalpha-Ostbeta mRNA was increased in cecum and proximal colon of cholic acid-fed mice and in chenodeoxycholate-treated mouse CT26 colon adenocarcinoma cells. Sequence analysis revealed potential cis-acting elements for farnesoid X receptor (FXR) and liver receptor homolog-1 (LRH-1) in the mouse Ostalpha and Ostbeta promoters and reporter constructs containing Ostalpha and Ostbeta 5'-flanking sequences were positively regulated by bile acids. Expression of a dominant-negative FXR, reduction of FXR with interfering small RNA (siRNA), or mutation of the potential FXR elements decreased Ostalpha and Ostbeta promoter activity and abolished the induction by chenodeoxycolic acid. Negative regulation of the Ostalpha and Ostbeta promoters by bile acids was mediated through LRH-1 elements. Ostalpha and Ostbeta promoter activities were increased by coexpression of LRH-1 and decreased by coexpression of SHP. Mutation of the potential LRH-1 elements and siRNA-mediated reduction of LRH-1 expression decreased basal promoter activity. As predicted from the promoter analyses, ileal Ostalpha and Ostbeta mRNA expressions were increased in wild-type mice administered the FXR agonist GW4064 and decreased in FXR-null mice. Immunoblotting analysis revealed that Ostalpha and Ostbeta intestinal protein expressions correlated with mRNA expression. The mouse Ostalpha and Ostbeta promoters are unusual in that they contain functional FXR and LRH elements, which mediate, respectively, positive and negative feedback regulation by bile acids. Although the positive regulatory pathway appears to be dominant, this arrangement provides a mechanism to finely titrate Ostalpha-Ostbeta expression to the bile acid flux. PMID- 16357059 TI - beta-Casomorphin-7 regulates the secretion and expression of gastrointestinal mucins through a mu-opioid pathway. AB - We have recently shown that beta-casomorphin-7, a milk opioid peptide, strongly stimulates mucin secretion in the rat jejunum through a nervous pathway and opioid receptor activation. In this study, the hypothesis that beta-casomorphin-7 may also act directly on intestinal goblet cells was investigated in vitro in rat and human intestinal mucin-producing cells (DHE and HT29-MTX) using quantitative and semiquantitative RT-PCR and ELISA. The presence of mu-opioid receptors was demonstrated in rat goblet cells in the upper half of the colonic crypt and in the two cell lines by immunohistochemistry and RT-PCR. In rat DHE cells, beta casomorphin-7 increased the expression of rat mucin (rMuc)2 and rMuc3 but not rMuc1, rMuc4, and rMuc5AC. This effect was time and dose dependent, with the maximum of increase in transcripts being noticed for a concentration of 10(-4) M after 2 h of stimulation for rMuc2 (225% of controls) and 4 h of stimulation for rMuc3 (208% of controls). Mucin secretion was maximally increased after 8 h of stimulation. Interestingly, these effects were prevented by pretreatment of the cells with the mu-opioid antagonist cyprodime. In human HT29-MTX cells, beta casomorphin-7 (10(-4) M) also increased MUC5AC mRNA levels (219% after 24 h of stimulation) and the secretion of this mucin (169% of controls). In conclusion, beta-casomorphin-7 may contribute significantly to mucin production via a direct effect on intestinal goblet cells and the activation of mu-opioid receptors. Because intestinal mucins have a crucial mucosal protective function, dairy products containing beta-casomorphin-7 may improve intestinal protection and could have dietary and health applications. PMID- 16357061 TI - Effects of cholera toxin on the potential difference and motor responses induced by distension in the rat proximal small intestine in vivo. AB - Cholera toxin (CT) may induce uncontrolled firing in recurrent networks of secretomotor neurons in the submucous plexus. This hypothesis was tested in chloralose-anesthetized rats in vivo. The secretory reflex response to graded intestinal distension was measured with or without prior exposure to luminal CT. The transmural potential difference (PD) was used as a marker for electrogenic chloride secretion. In controls, distension increased PD, and this response was reduced by the neural blocker tetrodotoxin given serosally and the vasoactive intestinal peptide (VIP) receptor antagonist [4Cl-d-Phe(6),Leu(17)]VIP (2 mug.min(-1).kg(-1) iv) but unaffected by the serotonin 5-HT(3) receptor antagonist granisetron, by the nicotinic receptor antagonist hexamethonium, by the muscarinic receptor antagonist atropine, or by the cyclooxygenase inhibitor indomethacin. Basal PD increased significantly with time in CT-exposed segments, an effect blocked by granisetron, by indomethacin, and by [4Cl-d Phe(6),Leu(17)]VIP but not by hexamethonium or atropine. In contrast, once the increased basal PD produced by CT was established, [4Cl-d-Phe(6),Leu(17)]VIP and indomethacin had no significant effect, whereas granisetron and hexamethonium markedly depressed basal PD. CT significantly reduced the increase in PD produced by distension, an effect reversed by granisetron, indomethacin, and atropine. CT also activated a specific motility response to distension, repeated cluster contractions, but only in animals pretreated with granisetron, indomethacin, or atropine. These data are compatible with the hypothesis that CT induces uncontrolled activity in submucous secretory networks. Development of this state depends on 5-HT(3) receptors, VIP receptors, and prostaglandin synthesis, whereas its maintenance depends on 5-HT(3) and nicotinic receptors but not VIP receptors. The motility effects of CT (probably reflecting myenteric activity) are partially suppressed via a mechanism involving 5-HT(3) and muscarinic receptors and prostaglandin synthesis. PMID- 16357060 TI - Dystroglycan receptor is involved in integrin activation in intestinal epithelia. AB - The dystroglycans (alpha-DG and beta-DG), which play important roles in the formation of basement membranes, have been well studied in skeletal muscle and nerve, but their expression and localization in intestinal epithelial cells has not been previously investigated. Here, we demonstrated that the DG complex, composed of alpha-DG, beta-DG, and utrophin, is specifically expressed in the basolateral membrane of the Caco-2-BBE monolayer. The DG complex coprecipitated with beta(1)-integrin, suggesting a possible interaction among these proteins. In addition, we observed that activation of DG receptors by laminin-1 enhanced the interaction between beta(1)-integrin and laminin-1, whereas activation of DG receptors by laminin-2 reduced the interaction between beta(1)-integrin and laminin-2. Finally, we demonstrated that the intracellular COOH-terminal tail of beta-DG and its binding to the DG binding domain of utrophin are crucial for the interactions between laminin-1/-2 and beta(1)-integrin. Collectively, these novel results indicate that dystroglycans play important roles in the regulation of interactions between intestinal epithelial cells and the extracellular matrix. PMID- 16357062 TI - Heregulin-alpha and heregulin-beta expression is linked to a COX-2-PGE2 pathway in human gastric fibroblasts. AB - We have previously shown heregulin (HRG)-alpha expression in human gastric fibroblasts and its stimulation of gastric epithelial cell growth. Although cyclooxygenase (COX)-2 has also been shown to stimulate growth factor production in these cells, the interaction between COX-2 and HRG remains unknown. Conditioned media (CM) from gastric fibroblasts incubated with PGE(2) or interleukin (IL)-1beta, a well known COX-2 inducer, were analyzed for their effect on erbB3 tyrosine phosphorylation in MKN28 gastric epithelial cells. HRG protein expression in fibroblast lysates and CM was also examined by western blot. HRG-alpha and HRG-beta mRNA expression in gastric fibroblasts and human gastric tissue was examined by real-time quantitative PCR. HRG and COX-2 expressions in surgical resections of human gastric ulcer tissue were examined immunohistochemically. CM from fibroblasts incubated with PGE(2), or IL-1beta, stimulated erbB3 phosphorylation in MKN28 cells. Preincubation of the fibroblasts with celecoxib, a selective COX-2 inhibitor, suppressed CM-induced erbB3 phosphorylation. This inhibition was reversed by exogenous PGE(2). As with erbB3 phophorylation, IL-1beta stimulated both HRG-alpha and HRG-beta mRNA expression, as well as HRG release into gastric fibroblast CM. IL-1beta-stimulated HRG expression and release were also inhibited by celecoxib, and exogenous PGE(2) restored this inhibitory effect, suggesting the activation of an IL-1beta-COX-2 PGE(2) pathway that culminates in the release of HRG from fibroblasts. HRG-alpha and HRG-beta mRNA levels were significantly higher in gastric ulcer tissue than in normal gastric mucosa. HRG immunoreactivity was found in interstitial cells of the gastric ulcer bed and coexpressed with COX-2. These results suggest that HRG might be a new member of the growth factor family involved in the COX-2-dependent ulcer repair process. PMID- 16357063 TI - Yin yang 1 (YY1) represses histidine decarboxylase gene expression with SREBP-1a in part through an upstream Sp1 site. AB - Histidine decarboxylase (HDC) is the enzyme that converts histidine to histamine, a bioamine that plays an important role in many physiological aspects including allergic responses, inflammation, neurotransmission, and gastric acid secretion. In previous studies, we demonstrated that Kruppel-like factor 4 represses HDC promoter activity in a gastric cell line through both an upstream Sp1-binding GC box (GGGCGG sequence) and downstream gastrin-responsive elements. In the current study, Yin Yang 1 (YY1), a pleiotropic transcriptional factor, was also shown in cotransfection assays to repress HDC promoter activity through the upstream GC box. DNA affinity purification assay demonstrated that YY1 was pulled down specifically by the upstream GC box. In addition, sterol-responsive element binding protein 1a (SREBP-1a), a transcriptional factor that binds YY1, represses the HDC promoter. Interestingly, deletion analysis and cotransfection assays indicated that mutation of the upstream GC box or truncation of downstream gastrin-responsive elements in the HDC promoter disrupted the inhibitory effect of YY1 and SREBP-1a in an identical fashion. Furthermore, quantitative real-time PCR analysis indicated that gastrin treatment downregulated SREBP-1a gene expression and reduced the DNA binding activity of SREBP in EMSAs. Taken together, these results suggest that YY1 and SREBP-1a form a complex to inhibit HDC gene expression through both the upstream GC box and downstream gastrin responsive elements and gastrin-induced activation of HDC gene expression is mediated at least partly through downregulation of transcriptional repressors such as SREBPs. PMID- 16357064 TI - Saturated, but not n-6 polyunsaturated, fatty acids induce insulin resistance: role of intramuscular accumulation of lipid metabolites. AB - Consumption of a Western diet rich in saturated fats is associated with obesity and insulin resistance. In some insulin-resistant phenotypes this is associated with accumulation of skeletal muscle fatty acids. We examined the effects of diets high in saturated fatty acids (Sat) or n-6 polyunsaturated fatty acids (PUFA) on skeletal muscle fatty acid metabolite accumulation and whole-body insulin sensitivity. Male Sprague-Dawley rats were fed a chow diet (16% calories from fat, Con) or a diet high (53%) in Sat or PUFA for 8 wk. Insulin sensitivity was assessed by fasting plasma glucose and insulin and glucose tolerance via an oral glucose tolerance test. Muscle ceramide and diacylglycerol (DAG) levels and triacylglycerol (TAG) fatty acids were also measured. Both high-fat diets increased plasma free fatty acid levels by 30%. Compared with Con, Sat-fed rats were insulin resistant, whereas PUFA-treated rats showed improved insulin sensitivity. Sat caused a 125% increase in muscle DAG and a small increase in TAG. Although PUFA also resulted in a small increase in DAG, the excess fatty acids were primarily directed toward TAG storage (105% above Con). Ceramide content was unaffected by either high-fat diet. To examine the effects of fatty acids on cellular lipid storage and glucose uptake in vitro, rat L6 myotubes were incubated for 5 h with saturated and polyunsaturated fatty acids. After treatment of L6 myotubes with palmitate (C16:0), the ceramide and DAG content were increased by two- and fivefold, respectively, concomitant with reduced insulin stimulated glucose uptake. In contrast, treatment of these cells with linoleate (C18:2) did not alter DAG, ceramide levels, and glucose uptake compared with controls (no added fatty acids). Both 16:0 and 18:2 treatments increased myotube TAG levels (C18:2 vs. C16:0, P < 0.05). These results indicate that increasing dietary Sat induces insulin resistance with concomitant increases in muscle DAG. Diets rich in n-6 PUFA appear to prevent insulin resistance by directing fat into TAG, rather than other lipid metabolites. PMID- 16357065 TI - Dual-wavelength phosphorimetry for determination of cortical and subcortical microvascular oxygenation in rat kidney. AB - This study presents a dual-wavelength phosphorimeter developed to measure microvascular PO2 (microPO2) in different depths in tissue and demonstrates its use in rat kidney. The used phosphorescent dye is Oxyphor G2 with excitation bands at 440 and 632 nm. The broad spectral gap between the excitation bands combined with a relatively low light absorption of 632 nm light by tissue results in a marked difference in penetration depths of both excitation wavelengths. In rat kidney, we determine the catchments depth of the 440-nm excitation to be 700 microm, whereas the catchments depth of 632 nm is as much as 4 mm. Therefore, the measurements differentiate between cortex and outer medulla, respectively. In vitro, no difference in PO2 readings between both channels was found. On the rat kidney in vivo, the measured cortical microPO2 was on average 20 Torr higher than the medullary microPO2 over a wide PO2 range induced by variations in inspired oxygen fraction. Examples provided from endotoxemia and resuscitation show differences in responses of mean cortical and medullary PO2 readings as well as in the shape of the PO2 histograms. It can be concluded that oxygen-dependent quenching of phosphorescence of Oxyphor G2 allows quantitative measurement of microPO2 noninvasively in two different depths in vivo. Oxygen levels measured by this technique in the rat renal cortex and outer medulla are consistent with previously published values detected by Clark-type oxygen electrodes. Dual wavelength phosphorimetry is excellently suited for monitoring microPO2 changes in two different anatomical layers under pathophysiological conditions with the characteristics of providing oxygen histograms from two depths and having a penetration depth of several millimeters. PMID- 16357067 TI - Quantitative interrelationship between Gibbs-Donnan equilibrium, osmolality of body fluid compartments, and plasma water sodium concentration. AB - The presence of negatively charged, impermeant proteins in the plasma space alters the distribution of diffusible ions in the plasma and interstitial fluid (ISF) compartments to preserve electroneutrality. We have derived a new mathematical model to define the quantitative interrelationship between the Gibbs Donnan equilibrium, the osmolality of body fluid compartments, and the plasma water Na+ concentration ([Na+]pw) and validated the model using empirical data from the literature. The new model can account for the alterations in all ionic concentrations (Na+ and non-Na+ ions) between the plasma and ISF due to Gibbs Donnan equilibrium. In addition to the effect of Gibbs-Donnan equilibrium on Na+ distribution between plasma and ISF, our model predicts that the altered distribution of osmotically active non-Na+ ions will also have a modulating effect on the [Na+]pw by affecting the distribution of H2O between the plasma and ISF. The new physiological insights provided by this model can for the first time provide a basis for understanding quantitatively how changes in the plasma protein concentration modulate the [Na+]pw. Moreover, this model defines all known physiological factors that may modulate the [Na+]pw and is especially helpful in conceptually understanding the pathophysiological basis of the dysnatremias. PMID- 16357066 TI - Effect of a short-term diet and exercise intervention on oxidative stress, inflammation, MMP-9, and monocyte chemotactic activity in men with metabolic syndrome factors. AB - The present study was designed to examine the effects of lifestyle modification on key contributing factors to atherogenesis, including oxidative stress, inflammation, chemotaxis, and cell adhesion. Obese men (n = 31), 15 of whom had metabolic syndrome, were placed on a high-fiber, low-fat diet in a 3-wk residential program where food was provided ad libitum and daily aerobic exercise was performed. In each subject, pre- and postintervention fasting blood was drawn for circulating levels of serum lipids, glucose and insulin (for estimation of insulin sensitivity), oxidative stress-generating enzyme myeloperoxidase and marker 8-isoprostaglandin F2alpha, the inflammatory protein C-reactive protein, soluble ICAM-1 as an indicator of endothelial activation, sP-selectin as a marker of platelet activation, the chemokine macrophage inflammatory protein-1alpha, and total matrix metalloproteinase-9. Using subject sera and human aortic endothelial cell culture systems, we measured VCAM-1 cell surface abundance and monocyte chemotactic protein-1, nitric oxide, superoxide, and hydrogen peroxide production in vitro by fluorometric detection. Also determined in vitro was serum-induced, monocyte adhesion and monocyte chemotactic activity. After 3 wk, significant reductions (P < 0.05) in body mass index, all serum lipids and lipid ratios, fasting glucose, insulin, homeostasis model assessment for insulin resistance, myeloperoxidase, 8-isoprostaglandin F2alpha, C-reactive protein, soluble ICAM-1, soluble P-selectin, macrophage inflammatory protein-1alpha, and matrix metalloproteinase-9 were noted. In vitro, serum-stimulated cellular VCAM-1 expression, monocyte chemotactic protein-1 production, and fluorometric detection of superoxide and hydrogen peroxide production decreased, whereas a concomitant increase in NO production was noted (all P < 0.01). Additionally, both monocyte adhesion (P < 0.05) and MCA (P < 0.01) decreased. Nine of 15 were no longer positive for metabolic syndrome postintervention. Intensive lifestyle modification may ameliorate novel coronary artery disease risk factors in men with metabolic syndrome factors before reversal of obesity. PMID- 16357068 TI - Heat intolerance: does gene transcription contribute? AB - During exertion in the heat, heat-intolerant (HI) subjects have a physiological disability in metabolic heat dissipation. The HI state is either permanent or temporary, depending on whether it stems from transient predisposing factors or inherent thermoregulatory dysfunction. In this investigation, we studied protein levels of heat shock protein (HSP) 70 and HSP72, HSP90, bcl-2xL, glutathione S transferase-p, heat shock factor-1, TATA-binding protein-associated factor, and NF-kappaB transcripts using Western blot and quantitative RT-PCR, respectively, in lymphocytes of HI and tolerant (T) male volunteers of similar anthropometric features. Measurements were made from blood drawn before, during the heat tolerance test (3.5 mph, 40 degrees C, 40% relative humidity, 2 h), and 1 h after recovery at 24 degrees C. Rectal and skin temperatures, as well as heart rate, were continuously recorded. Of 58 subjects, 7 were identified as HI, with a significantly higher physiological strain index than in the T group (6.3 +/- 0.9 vs. 3.8 +/- 0.6, P < 0.001). The responsiveness of the vasculature to thermal stimuli was decreased in the HI group, as indicated by rectal temperature minus skin temperature. The HSP72 level in the HI group dropped during the recovery session (P < 0.01), whereas that of the T group continued to rise. A significantly increased expression of the transcription factors in the T subjects and significantly decreased expression in the HI group (P < 0.009, 0.013, and 0.005 for heat shock factor-1, NF-kappaB, and TATA-binding protein-associated factor, respectively) points to impaired transcriptional processes in the HI group. Our data suggest that transcriptional malfunction and sluggishness of the vasculature to thermal stimuli are predisposing factors in the HI group. PMID- 16357069 TI - Effect of voluntary exercise on peripheral tissue glucocorticoid receptor content and the expression and activity of 11beta-HSD1 in the Syrian hamster. AB - Recent findings indicate that elevated levels of glucocorticoids (GC), governed by the expression of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) and GC receptors (GR), in visceral adipose tissue and skeletal muscle lead to increased insulin resistance and the metabolic syndrome. Paradoxically, evidence indicates that aerobic exercise attenuates the development of the metabolic syndrome even though it stimulates acute increases in circulating GC levels. To investigate the hypothesis that training alters peripheral GC action to maintain insulin sensitivity, young male hamsters were randomly divided into sedentary (S) and trained (T) groups (n = 8 in each). The T group had 24-h access to running wheels over 4 wk of study. In muscle, T hamsters had lower 11beta-HSD1 protein expression (19.2 +/- 1.40 vs. 22.2 +/- 0.96 optical density, P < 0.05), similar 11beta-HSD1 enzyme activity (0.9 +/- 0.27% vs. 1.1 +/- 0.26), and lower GR protein expression (9.7 +/- 1.86 vs. 15.1 +/- 1.78 optical density, P < 0.01) than S hamsters. In liver, 11beta-HSD1 protein expression tended to be lower in T compared with S (19.2 +/- 0.56 vs. 21.4 +/- 1.05, P = 0.07), whereas both enzyme activity and GR protein expression were similar. In contrast, visceral adipose tissue contained approximately 2.7-fold higher 11beta-HSD1 enzyme activity in T compared with S (12.9 +/- 3.3 vs. 4.8 +/- 1.5% conversion, P < 0.05) but was considerably smaller in mass (0.24 +/- 0.02 vs. 0.71 +/- 0.06 g). Thus the intracellular adaptation of GC regulators to exercise is tissue specific, resulting in decreases in GC action in skeletal muscle and increases in GC action in visceral fat. These adaptations may have important implications in explaining the protective effects of aerobic exercise on insulin resistance and other symptoms of the metabolic syndrome. PMID- 16357070 TI - Nitric oxide regulation of microvascular oxygen exchange during hypoxia and hyperoxia. AB - The objective of this work was to test the hypothesis that the limitation of nitric oxide (NO) availability accentuates microvascular reactivity to oxygen. The awake hamster chamber window model was rendered hypoxic and hyperoxic by ventilation with 10 and 100% oxygen. Systemic and microvascular parameters were determined in the two conditions and compared with normoxia in a group receiving the NO scavenger nitronyl nitroxide and a control group receiving only the vehicle (saline). Mean arterial blood pressure did not change with different gas mixtures during infusion of the vehicle, but it increased significantly in the NO depleted group. NO scavenging increased the reactivity of microvessels to the changed oxygen supply, causing the arteriolar wall to significantly increase oxygen consumption. Tissue Po2 was correspondingly significantly reduced during NO scavenger infusion. The present findings support the hypothesis that microvascular oxygen consumption is proportional to oxygen-induced vasoconstriction. The effect of oxygen on vascular tone is modulated by NO. As a consequence, NO acts as a regulator of the vessel wall oxygen consumption. The vessel wall consumes oxygen in proportion to the local Po2, and an impairment of NO availability renders the circulation more sensitive to changes in the oxygen supply. PMID- 16357071 TI - Chronic intermittent hypoxia alters NE reactivity and mechanics of skeletal muscle resistance arteries. AB - Although arterial dilator reactivity is severely impaired during exposure of animals to chronic intermittent hypoxia (CIH), few studies have characterized vasoconstrictor responsiveness in resistance arteries of this model of sleep disordered breathing. Sprague-Dawley rats were exposed to CIH (10% inspired O2 fraction for 1 min at 4-min intervals; 12 h/day) for 14 days. Control rats were housed under normoxic conditions. Diameters of isolated gracilis muscle resistance arteries (GA; 120-150 microm) were measured by television microscopy before and during exposure to norepinephrine (NE) and angiotensin II (ANG II) and at various intraluminal pressures between 20 and 140 mmHg in normal and Ca2+-free physiological salt solution. There was no difference in the ability of GA to constrict in response to ANG II (P = 0.42; not significant; 10(-10)-10(-7) M). However, resting tone, myogenic activation, and vasoconstrictor responses to NE (P < 0.001; 10(-9)-10(-6) M) were reduced in CIH vs. controls. Treatment of rats with the superoxide scavenger 4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl (tempol; 1 mM) in the drinking water restored myogenic responses and NE-induced constrictions of CIH rats, suggesting that elevated superoxide production during exposure to CIH attenuates vasoconstrictor responsiveness to NE and myogenic activation in skeletal muscle resistance arteries. CIH also leads to an increased stiffness and reduced vessel wall distensibility that were not correctable with oral tempol treatment. PMID- 16357072 TI - O-GlcNAc level variations are associated with the development of skeletal muscle atrophy. AB - O-linked N-acetylglucosaminylation (O-GlcNAc) is a regulatory posttranslational modification of nucleocytoplasmic proteins, which consists of the attachment of N acetylglucosamine to serine or threonine residues of a protein. This glycosylation is a ubiquitous posttranslational modification, which probably plays important roles in many aspects of protein function. Our laboratory has previously reported that, in skeletal muscle, proteins of the glycolytic pathway and energetic metabolism and contractile proteins were O-GlcNAc modified (Cieniewski-Bernard C, Bastide B, Lefebvre T, Lemoine J, Mounier Y, and Michalski JC. Mol Cell Proteomics 3: 577-585, 2004). O-GlcNAc has been recently demonstrated to play a role in modulating cellular function in response to nutrition and also in stress conditions. Therefore, we have investigated here the implication of the glycosylation/deglycosylation process in the development of atrophy in rat skeletal muscle after hindlimb unloading. The high O-GlcNAc level found in control soleus [compared with control extensor digitorum longus (EDL)] becomes lower in atrophied soleus. On the opposite side, the low rate of O-GlcNAc in control EDL reaches higher levels in EDL, not atrophied after hindlimb unloading. These variations in O-GlcNAc level are correlated with a variation of the O-GlcNAc process enzyme activities and could be associated with a differential expression of heat shock proteins. Our results suggest that O-GlcNAc variations could control the muscle protein homeostasis and be implicated in the regulation of muscular atrophy. PMID- 16357073 TI - Constitutive pro- and anti-inflammatory cytokine and growth factor response to exercise in leukocytes. AB - Leukocytosis following exercise is a well-described phenomenon of stress/inflammatory activation in healthy humans. We hypothesized that, despite this increase in circulating inflammatory cells, exercise would paradoxically induce expression of both pro- and anti-inflammatory cytokines and growth factors within these cells. To test this hypothesis, 11 healthy adult men, 18-30 yr old, performed a 30-min bout of heavy cycling exercise; blood sampling was at baseline, end-exercise, and 60 min into recovery. The percentage of leukocytes positive for intracellular cytokines and growth factors and mean fluorescence intensity was obtained by flow cytometry. Proinflammatory cytokines (IL-1alpha, IL-2, IFN-gamma, and TNF-alpha), a pleiotropic cytokine (IL-6), and anti inflammatory cytokines and growth factors [IL-4, IL-10, growth hormone (GH), and IGF-I] were examined. Median fluorescence intensity was not affected by exercise; however, we found a number of significant changes (P < 0.05 by mixed linear model and modified t-test) in the numbers of circulating cells positive for particular mediators. The pattern of expression reflected both pro- and anti-inflammatory functions. In T-helper lymphocytes, TNF-alpha, but also IL-6, and IL-4 were significantly increased. In monocytes, both IFN-gamma and IL-4 increased. B lymphocytes positive for GH and IGF-I increased significantly. GH-positive granulocytes also significantly increased. Collectively, these observations indicate that exercise primes an array of pro- and anti-inflammatory and growth factor expression within circulating leukocytes, perhaps preparing the organism to effectively respond to a variety of stressors imposed by exercise. PMID- 16357074 TI - Growth of the small airways and alveoli from childhood to the adult lung measured by aerosol-derived airway morphometry. AB - Understanding the human development of pulmonary air spaces is important for calculating the dose from exposure to inhaled materials as a function of age. We have measured, in vivo, the air space caliber of the small airways and alveoli at their natural full distension [total lung capacity (TLC)] by aerosol-derived airway morphometry in 53 children of age 6-22 yr and 59 adults of age 23-80 yr. Aerosol-derived airway morphometry utilizes the gravitational settling time of inhaled inert particles to infer the vertical distance necessary to produce the observed loss of particles to the airway surfaces at sequential depths into the lung. Previously, we identified anatomical features of the lung: the caliber of the transitional bronchioles [transitional effective air space dimension (EADtrans)]; the mean linear dimension of the alveoli (EADmin); and a measure of conducting airway volume [volumetric lung depth (VLDtrans)]. In the present study, we found that EADmin increased with age, from 184 microm at age 6 to 231 microm at age 22, generally accounting for the increase in TLC observed over this age range. EADtrans did not increase with TLC, averaging 572 microm, but increased with subject age and height when the entire age range of 6-80 yr is included {EADtrans (microm)=0.012[height (cm)]x[age (yr)]+508; P=0.007}. VLDtrans scaled linearly with lung volume, but VLDtrans relative to TLC did not change with age, averaging 7.04+/-1.55% of TLC. The data indicate that from childhood (age of 6 yr) to adulthood a constant number of respiratory units is maintained while both the smallest bronchioles and alveoli expand in size to produce the increased lung volume with increased age and height. PMID- 16357075 TI - Dead space: simplicity to complexity. PMID- 16357076 TI - Regulation of the cerebral circulation. PMID- 16357077 TI - Extraluminal tissue pressure: what does it mean? PMID- 16357078 TI - "Fat adaptation" for athletic performance: the nail in the coffin? PMID- 16357079 TI - Why do we have both peripheral and central chemoreceptors? PMID- 16357080 TI - Is obstructive sleep apnea the cause of sympathetic nervous activation in human obesity? PMID- 16357081 TI - Increased spinal reflex excitability is not associated with neural plasticity underlying the cross-education effect. AB - The purpose of this study was to examine the effects of a 5-wk unilateral, isometric strength-training program on plasticity in the spinal Hoffmann (H-) reflex in both the trained and untrained legs. Sixteen participants, 22-42 yr old, were assigned to either a control (n = 6) or an exercise group (n = 10). Both groups were tested for plantar flexion maximal voluntary isometric contractions (MVIC) and soleus H-reflex amplitude in both limbs, at the beginning and at the end of a 5-wk interval. Participants in the exercise group showed significantly increased MVIC in both legs after training (P < 0.05), whereas strength was unchanged in the control group for either leg. Subjects in the exercise group displayed increased (P < 0.05) H-reflex amplitudes on the ascending limb of the recruitment curve (at an equivalent M wave of 5% of the maximal M wave, H(A)) only in the trained leg. Maximal H-reflex and M-wave remained unchanged with training. Increased amplitude of H(A) in the trained limb concurrent with increased strength suggests that spinal mechanisms may underlie the changes in strength, possibly because of increased alpha-motoneuronal excitability or reduced presynaptic inhibition. Despite a similar increase in strength in the contralateral limb of the exercise group, H(A) amplitude was unchanged. We conclude that the cross-education effect of strength training may be due to supraspinal to a greater extent than spinal mechanisms. PMID- 16357082 TI - Effect of chronic hypoxia on purinergic synaptic transmission in rat carotid body. AB - Recent studies indicate that chemoafferent nerve fiber excitation in the rat carotid body is mediated by acetylcholine and ATP, acting at nicotinic cholinergic receptors and P2X2 purinoceptors, respectively. We previously demonstrated that, after a 10- to 14-day exposure to chronic hypoxia (CH), the nicotinic cholinergic receptor blocker mecamylamine no longer inhibits rat carotid sinus nerve (CSN) activity evoked by an acute hypoxic challenge. The present experiments examined the effects of CH (9-16 days at 380 Torr) on the expression of P2X2 purinoceptors in carotid body and chemoafferent neurons, as well as the effectiveness of P2X2 receptor blocking drugs on CSN activity evoked by hypoxia. In the normal carotid body, immunocytochemical studies demonstrated a dense plexus of P2X2-positive nerve fibers penetrating lobules of type I cells. In addition, type I cells were lightly stained, indicating P2X2 receptor expression. After CH, the intensity of P2X2 receptor immunostaining was maintained in chemosensory type I cells and in the soma of chemoafferent neurons. P2 receptor expression on type I cells was confirmed by demonstrations of ATP evoked increased intracellular Ca2+; this response was modulated by simultaneous exposure to hypoxia. In normal preparations, CSN activity evoked by hypoxia in vitro was 65% inhibited in the presence of specific P2X2 receptor antagonists. However, unlike the absence of mecamylamine action after CH, P2X2 antagonists remained effective against hypoxia-evoked activity after CH. Our findings indicate that ATP acting at P2X2 receptors contributes to adjusted chemoreceptor activity after CH, indicating a possible role for purinergic mechanisms in the adaptation of the carotid body in a chronic low-O2 environment. PMID- 16357083 TI - Time course of COX-1 and COX-2 expression during ischemia-reperfusion in rat skeletal muscle. AB - The aim of this study was to assess cyclooxygenase (COX)-1 and COX-2 expression in skeletal muscle after an ischemia-reperfusion (I/R). Male Sprague-Dawley rats were subjected to unilateral hindlimb ischemia for 2 h and then euthanized after 0, 1, 2, 4, 6, 10, 24, and 72 h of reperfusion. The COX protein and mRNA were assessed in control and injured gastrocnemius muscle. Muscle damage was indirectly determined by plasma creatine kinase activity and edema by weighing wet muscle. Creatine kinase activity in plasma increased as early as 1 h after reperfusion and returned to control levels by 72 h of reperfusion. Edema was observed at 6 and 10 h of reperfusion, but histological investigations showed an absence of tissular inflammatory cell infiltration. COX-1 mRNA was expressed in control muscle and was increased at 72 h of reperfusion, but the levels of associated COX-1 protein detected in control and injured gastrocnemius muscle were similar. COX-2 mRNA was not, or only slightly, detectable in control muscle and after I/R. In contrast, I/R induced major overexpression of COX-2 immunoreactivity at 6 and 10 h of reperfusion with a maximum at 10 h, whereas COX 2 protein was undetectable in control muscle. In conclusion, hindlimb I/R induced a large overexpression of COX-2 but not COX-1 protein between 6 and 10 h after injury. These results suggest a role for COX-2 enzyme in such pathophysiological conditions of the skeletal muscle. PMID- 16357085 TI - Endothelial influences on cerebrovascular tone. AB - The cerebrovascular endothelium exerts a profound influence on cerebral vessels and cerebral blood flow. This review summarizes current knowledge of various dilator and constrictor mechanisms intrinsic to the cerebrovascular endothelium. The endothelium contributes to the resting tone of cerebral arteries and arterioles by tonically releasing nitric oxide (NO*). Dilations can occur by stimulated release of NO*, endothelium-derived hyperpolarization factor, or prostanoids. During pathological conditions, the dilator influence of the endothelium can turn to that of constriction by a variety of mechanisms, including decreased NO* bioavailability and release of endothelin-1. The endothelium may participate in neurovascular coupling by conducting local dilations to upstream arteries. Further study of the cerebrovascular endothelium is critical for understanding the pathogenesis of a number of pathological conditions, including stroke, traumatic brain injury, and subarachnoid hemorrhage. PMID- 16357084 TI - Role of astrocytes in cerebrovascular regulation. AB - Astrocytes send processes to synapses and blood vessels, communicate with other astrocytes through gap junctions and by release of ATP, and thus are an integral component of the neurovascular unit. Electrical field stimulations in brain slices demonstrate an increase in intracellular calcium in astrocyte cell bodies transmitted to perivascular end-feet, followed by a decrease in vascular smooth muscle calcium oscillations and arteriolar dilation. The increase in astrocyte calcium after neuronal activation is mediated, in part, by activation of metabotropic glutamate receptors. Calcium signaling in vitro can also be influenced by adenosine acting on A2B receptors and by epoxyeicosatrienoic acids (EETs) shown to be synthesized in astrocytes. Prostaglandins, EETs, arachidonic acid, and potassium ions are candidate mediators of communication between astrocyte end-feet and vascular smooth muscle. In vivo evidence supports a role for cyclooxygenase-2 metabolites, EETs, adenosine, and neuronally derived nitric oxide in the coupling of increased blood flow to increased neuronal activity. Combined inhibition of the EETs, nitric oxide, and adenosine pathways indicates that signaling is not by parallel, independent pathways. Indirect pharmacological results are consistent with astrocytes acting as intermediaries in neurovascular signaling within the neurovascular unit. For specific stimuli, astrocytes are also capable of transmitting signals to pial arterioles on the brain surface for ensuring adequate inflow pressure to parenchymal feeding arterioles. Therefore, evidence from brain slices and indirect evidence in vivo with pharmacological approaches suggest that astrocytes play a pivotal role in regulating the fundamental physiological response coupling dynamic changes in cerebral blood flow to neuronal synaptic activity. Future work using in vivo imaging and genetic manipulation will be required to provide more direct evidence for a role of astrocytes in neurovascular coupling. PMID- 16357086 TI - Neurovascular coupling in the normal brain and in hypertension, stroke, and Alzheimer disease. AB - The brain is critically dependent on a continuous supply of blood to function. Therefore, the cerebral vasculature is endowed with neurovascular control mechanisms that assure that the blood supply of the brain is commensurate to the energy needs of its cellular constituents. The regulation of cerebral blood flow (CBF) during brain activity involves the coordinated interaction of neurons, glia, and vascular cells. Thus, whereas neurons and glia generate the signals initiating the vasodilation, endothelial cells, pericytes, and smooth muscle cells act in concert to transduce these signals into carefully orchestrated vascular changes that lead to CBF increases focused to the activated area and temporally linked to the period of activation. Neurovascular coupling is disrupted in pathological conditions, such as hypertension, Alzheimer disease, and ischemic stroke. Consequently, CBF is no longer matched to the metabolic requirements of the tissue. This cerebrovascular dysregulation is mediated in large part by the deleterious action of reactive oxygen species on cerebral blood vessels. A major source of cerebral vascular radicals in models of hypertension and Alzheimer disease is the enzyme NADPH oxidase. These findings, collectively, highlight the importance of neurovascular coupling to the health of the normal brain and suggest a therapeutic target for improving brain function in pathologies associated with cerebrovascular dysfunction. PMID- 16357087 TI - Effects of nasal continuous positive airway pressure and oxygen supplementation on norepinephrine kinetics and cardiovascular responses in obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is characterized by noradrenergic activation. Nasal continuous positive airway pressure (CPAP) is the treatment of choice and has been shown to effectively reduce elevated norepinephrine (NE) levels. This study examined whether the reduction in NE after CPAP is due to an increase in NE clearance and/or a decrease of NE release rate. Fifty CPAP-naive OSA patients with an apnea-hypopnea index >15 were studied. NE clearance and release rates, circulating NE levels, urinary NE excretion, and blood pressure and heart rate were determined before and after 14 days of CPAP, placebo CPAP (CPAP administered at ineffective pressure), or oxygen supplementation. CPAP led to a significant increase in NE clearance (P < or = 0.01), as well as decreases in plasma NE levels (P < or = 0.018) and daytime (P < 0.001) and nighttime (P < 0.05) NE excretion. NE release rate was unchanged with treatment. Systolic (P < or = 0.013) and diastolic (P < or = 0.026) blood pressure and heart rate (P < or = 0.014) were decreased in response to CPAP but not in response to oxygen or placebo CPAP treatment. Posttreatment systolic blood pressure was best predicted by pretreatment systolic blood pressure and posttreatment NE clearance and release rate (P < 0.01). The findings indicate that one of the mechanisms through which CPAP reduces NE levels is through an increase in the clearance of NE from the circulation. PMID- 16357088 TI - Point: the muscle metaboreflex does restore blood flow to contracting muscles. PMID- 16357089 TI - Comments on Point-Counterpoint "Flow-mediated dilation does/does not reflect nitric oxide-mediated endothelial function". PMID- 16357090 TI - Comments on Point-Counterpoint "Positive effects of intermittent hypoxia (live high:train low) on exercise performance are/are not mediated primarily by augmented red cell volume". PMID- 16357091 TI - Comments on Point-Counterpoint "The muscle metaboreflex does/does not restore blood flow to contracting muscles". PMID- 16357092 TI - Comments on Women, hormones, and clinical trials: a beginning, not an end. PMID- 16357093 TI - Agonist and hypertonic saline-induced trafficking of the NK3-receptors on vasopressin neurons within the paraventricular nucleus of the hypothalamus. AB - The neurokinin 3 receptor (NK3R) is colocalized with vasopressinergic neurons within the hypothalamic paraventricular nucleus (PVN) and intraventricular injections of NK3R agonists stimulate vasopressin (VP) release. Our objectives were to test the hypotheses that intraventricular injections of the selective NK3R agonist, succinyl-[Asp6, N-Me-Phe8] substance P (senktide), activate NK3R expressed by vasopressinergic neurons within the PVN, and see whether NK3R expressed by vasopressinergic neurons in the PVN are activated by hyperosmolarity. NK3R internalization was used as a marker of receptor activation. Immunohistochemistry revealed that NK3Rs were membrane-bound on VP immunoreactive neurons in control rats. Following senktide injection, there was a significant increase in the appearance of NK3R immunoreactivity within the cytoplasm and a morphological rearrangement of the dendrites, indicating receptor internalization, which was reversible. Furthermore, pretreatment with a selective NK3R antagonist, SB-222200, blocked the senktide-induced VP release and internalization of the NK3R in the PVN. These results show that the trafficking of the NK3R is due to ligand binding the NK3R. In a subsequent experiment, rats were administered intragastric loads of 2 or 0.15 M NaCl, and NK3R immunohistochemistry was used to track activation of the receptor. In contrast to control rats, 2 M NaCl significantly increased plasma VP levels and caused the internalization of the NK3R on VP neurons. Also, NK3R immunoreactivity was located in the nuclei of vasopressinergic neurons after senktide and 2 M NaCl treatment. These results show that hyperosmolarity stimulates the local release of an endogenous ligand in the PVN to bind to and activate NK3R on vasopressinergic neurons. PMID- 16357094 TI - Hemodynamic and neuroendocrine responses to changes in sodium intake in compensated heart failure. AB - Patients with untreated heart failure (HF) exhibit a blunted hemodynamic and neuroendocrine response to a high sodium intake, leading to excessive sodium and water retention. However, it is not known whether this is the case for patients with compensated HF receiving angiotensin-converting enzyme inhibitors and beta adrenoreceptor blockers. Therefore, we determined the hemodynamic and neuroendocrine responses to 1 wk of a low-sodium diet (70 mmol/day) and 1 wk of a high-sodium diet (250 mmol/day) in 12 HF patients and 12 age-matched controls in a randomized, balanced fashion. During steady-state conditions, hemodynamic and neuroendocrine examinations were performed at rest and during bicycle exercise. In seated HF patients, high sodium intake increased body weight (1.6 +/- 0.4%), plasma volume (9 +/- 2%), cardiac index (14 +/- 6%), and stroke volume index (21 +/- 5%), whereas mean arterial pressure was unchanged. Therefore, the total peripheral resistance decreased by 10 +/- 4%. Similar hemodynamic changes were observed during an incremental bicycle exercise test. Plasma concentrations of angiotensin II and norepinephrine were suppressed, whereas plasma pro-B-type natriuretic peptide remained unchanged. In conclusion, high sodium intake was tolerated without any excessive sodium and water retention in medically treated patients with compensated HF. The observation that high sodium intake improves cardiac performance, induces peripheral vasodilatation, and suppresses the release of vasoconstrictor hormones does not support the advice for HF patients to restrict dietary sodium. PMID- 16357095 TI - Studies of oxytocin and vasopressin gene expression in the rat hypothalamus using exon- and intron-specific probes. AB - To develop a comprehensive approach for the study of oxytocin (OT) and vasopressin (VP) gene expression in the rat hypothalamus, we first developed an intronic riboprobe to measure OT heteronuclear RNA (hnRNA) levels by in situ hybridization histochemistry (ISHH). Using this 84-bp riboprobe, directed against intron 2 of the OT gene, we demonstrate strong and specific signals in neurons confined to the supraoptic (SON) and paraventricular (PVN) nuclei of the rat hypothalamus. We used this new intronic OT probe, together with other well established intronic and exonic OT and VP probes, to reevaluate OT and VP gene expression in the hypothalamus under two classical physiological conditions, acute osmotic stimulation, and lactation. We found that magnocellular neurons in 7- to 8-day lactating female rats exhibit increased OT but not VP hnRNA. Since VP mRNA is increased during lactation, this suggests that decreased VP mRNA degradation during lactation may be responsible for this change. In contrast, whereas there was the expected large increase in VP hnRNA after acute salt loading, there was no change in OT hnRNA, suggesting that acute hyperosmotic stimuli produce increased VP but not OT gene transcription. Hence, the use of both exon- and intron-specific probes, which distinguish the changes in hnRNA and mRNA levels, respectively, can provide insight into the relative roles of transcription and mRNA degradation processes in changes in gene expression evoked by physiological stimuli. PMID- 16357096 TI - Ionic mechanisms of excitation-induced regulation of Na+-K+-ATPase mRNA expression in isolated rat EDL muscle. AB - This study investigated the effects of electrical stimulation on Na+-K+-ATPase isoform mRNA, with the aim to identify factors modulating Na+-K+-ATPase mRNA in isolated rat extensor digitorum longus (EDL) muscle. Interventions designed to mimic exercise-induced increases in intracellular Na+ and Ca2+ contents and membrane depolarization were examined. Muscles were mounted on force transducers and stimulated with 60-Hz 10-s pulse trains producing tetanic contractions three times at 10-min intervals. Ouabain (1.0 mM, 120 min), veratridine (0.1 mM, 30 min), and monensin (0.1 mM, 30 min) were used to increase intracellular Na+ content. High extracellular K+ (13 mM, 60 min) and the Ca2+ ionophore A-23187 (0.02 mM, 30 min) were used to induce membrane depolarization and elevated intracellular Ca2+ content, respectively. Muscles were analyzed for Na+-K+-ATPase alpha1-alpha3 and beta1-beta3 mRNA (real-time RT-PCR). Electrical stimulation had no immediate effect on Na+-K+-ATPase mRNA; however at 3 h after stimulation, it increased alpha1, alpha2, and alpha3 mRNA by 223, 621, and 892%, respectively (P = 0.010), without changing beta mRNA. Ouabain, veratridine, and monensin increased intracellular Na+ content by 769, 724, and 598%, respectively (P = 0.001) but did not increase mRNA of any isoform. High intracellular K+ concentration elevated alpha1 mRNA by 160% (P = 0.021), whereas A-23187 elevated alpha3 mRNA by 123% (P = 0.035) but reduced beta1 mRNA by 76% (P = 0.001). In conclusion, electrical stimulation induced subunit-specific increases in Na+-K+ ATPase mRNA in isolated rat EDL muscle. Furthermore, Na+-K+-ATPase mRNA appears to be regulated by different stimuli, including cellular changes associated with membrane depolarization and increased intracellular Ca2+ content but not increased intracellular Na+ content. PMID- 16357097 TI - Nitric oxide modulation of ET(B) receptor-induced vasopressin release by rat and mouse hypothalamo-neurohypophyseal explants. AB - Endothelin (ET) peptides stimulate vasopressin (AVP) secretion via ET(B) receptors at hypothalamic loci. Nitric oxide modulates the actions of ET in the cardiovascular system and also influences neurotransmission and specifically suppresses firing of magnocellular neurons. The purpose of these studies was to ascertain whether nitric oxide, generated in response to ET(B) receptor stimulation, buffers the stimulatory effect of ET and suppresses AVP release. Studies were performed using a pharmacological approach in hypothalamo neurohypophyseal explants from rats, and an alternative strategy using explants from mice with an inactivating mutation of neuronal NOS (nNOS-/-) and their wild type parent strain. Whole explants in standard culture or only the hypothalamus of compartmentalized explants was exposed to the ET(B) selective agonist, IRL 1620 (10(-13) to 10(-8) M). Rat and wild-type mouse explants displayed similar responses, although absolute basal release rates were higher from murine explants. Maximal AVP release at 0.1 nM IRL 1620 was 311 +/- 63 (rat) and 422 +/- 112% basal x explant(-1) x h(-1) (mouse). Sodium nitroprusside (SNP; 0.1 mM) suppressed maximal AVP release to basal values. N(omega)-nitro-L-arginine methyl ester (L-NAME, 0.1 microM), which did not itself stimulate AVP secretion, more than doubled the response to 1 pM IRL 1620, from 136 +/- 28 to 295 +/- 49% basal x explant(-1) x h(-1) (P < 0.05) by rat explants. Explants from wild-type mice responded similarly. Explants from nNOS-/- mice had higher basal AVP secretory rate in response to 1 pM IRL 1620: 271 +/- 48 compared with 150 +/- 24% basal x explant(-1) x h(-1) (P < 0.05) from wild-type murine explants. In the nNOS-/-, SNP suppressed stimulated release, and L-NAME exerted no additional stimulatory effect: 243 +/- 38% basal x explant(-1) x h(-1). Thus nitric oxide inhibits the AVP secretory response induced by ET(B) receptor activation within the hypothalamo-neurohypophyseal system and is generated primarily by the nNOS isoform. The modulation of AVP secretion by ET and also nitric oxide can take place independently from their effects on cerebral blood flow, systemic hemodynamics, or the arterial baroreflex. PMID- 16357098 TI - Altered mitochondrial apparent affinity for ADP and impaired function of mitochondrial creatine kinase in gluteus medius of patients with hip osteoarthritis. AB - The cellular energy metabolism in human musculus gluteus medius (MGM) under normal conditions and hip osteoarthritis (OA) was explored. The functions of oxidative phosphorylation and energy transport systems were analyzed in permeabilized (skinned) muscle fibers by oxygraphy, in relation to myosin heavy chain (MHC) isoform distribution profile analyzed by SDS-PAGE, and to creatine kinase (CK) and adenylate kinase (AK) activities measured spectrophotometrically in the intact muscle. The results revealed high apparent Km for ADP in regulation of respiration that decreased after addition of creatine in MGM of traumatic patients (controls). OA was associated with increased sensitivity of mitochondrial respiration to ADP, decreased total activities of AK and CK with major reduction in mi-CK fraction, and attenuated effect of creatine on apparent Km for ADP compared with control group. It also included a complete loss of type II fibers in a subgroup of patients with the severest disease grade. It is concluded that energy metabolism in MGM cells is organized into functional complexes of mitochondria and ATPases. It is suggested that because of degenerative remodeling occurring during development of OA, these complexes become structurally and functionally impaired, which results in increased access of exogenous ADP to mitochondria and dysfunction of CK-phosphotransfer system. PMID- 16357099 TI - Influence of the adenosine A1 receptor on blood pressure regulation and renin release. AB - The present study was performed to investigate the role of adenosine A1 receptors in regulating blood pressure in conscious mice. Adenosine A1-receptor knockout (A1R-/-) mice and their wild-type (A1R+/+) littermates were placed on standardized normal-salt (NS), high-salt (HS), or salt-deficient (SD) diets for a minimum of 10 days before telemetric blood pressure and urinary excretion measurements in metabolic cages. On the NS diet, daytime and nighttime mean arterial blood pressure (MAP) was 7-10 mmHg higher in A1R-/- than in A1R+/+ mice. HS diet did not affect the MAP in A1R-/- mice, but the daytime and nighttime MAP of the A1R+/+ mice increased by approximately 10 mmHg, to the same level as that in the A1R-/-. On the SD diet, day- and nighttime MAP decreased by approximately 6 mmHg in both A1R-/- and A1R+/+ mice, although the MAP remained higher in A1R-/- than in A1R+/+ mice. Although plasma renin levels decreased with increased salt intake in both genotypes, the A1R-/- mice had an approximately twofold higher plasma renin concentration on all diets compared with A1R+/+ mice. Sodium excretion was elevated in the A1R-/- compared with the A1R+/+ mice on the NS diet. There was no difference in sodium excretion between the two genotypes on the HS diet. Even on the SD diet, A1R-/- mice had an increased sodium excretion compared with A1R+/+ mice. An abolished tubuloglomerular feedback response and reduced tubular reabsorption can account for the elevated salt excretion found in A1R-/- animals. The elevated plasma renin concentrations found in the A1R-/- mice could also result in increased blood pressure. Our results confirm that adenosine, acting through the adenosine A1 receptor, plays an important role in regulating blood pressure, renin release, and sodium excretion. PMID- 16357100 TI - Prenatal exposure to interleukin-6 results in inflammatory neurodegeneration in hippocampus with NMDA/GABA(A) dysregulation and impaired spatial learning. AB - During pregnancy, infection or immune responses induce cytokine release, which might influence fetal neurodevelopment, leading to neurodegenerative disease in adulthood. Because the hippocampus is a key area for learning and memory, we evaluated 4- and 24-wk-old rats for the effects of early and late prenatal exposure to interleukin-6 (IL-6) on hippocampal morphology, expression of mRNA for IL-6, the gamma-aminobutyric acid receptor (GABA(Aalpha5)), the NR1 subunit of the N-methyl-D-aspartate receptor, and glial fibrillary acidic protein (GFAP), caspase-3 protein and mRNA levels, and learning abilities. Late exposure increased serum IL-6 and hippocampal expression of IL-6 mRNA at 4 and 24 wk. All adult rats showed neuronal loss in the hilus and astrogliosis; males had losses mainly in the CA2 and CA3 regions, and females in CA1. Expression of GABA(Aalpha5), NR1, and GFAP mRNA increased in late-exposed males and females at 4 and 24 wk. mRNA and protein levels of the apoptosis marker caspase-3 were increased in all late-exposed rats except males at 4 wk. Evaluation of hippocampus-dependent working memory in the Morris water maze at 20 wk of age showed increases in escape latency and time spent near the pool wall in all IL-6 adult rats, especially females. These findings suggest that fetal IL-6 exposure, especially in late pregnancy, leads to increased IL-6 levels in the circulation and hippocampus, abnormalities of hippocampal structural and morphology, and decreased learning during adulthood. PMID- 16357101 TI - Corticotropin-releasing hormone in the lateral parabrachial nucleus inhibits sodium appetite in rats. AB - The present study investigated the role of corticotropin-releasing hormone (CRH) in the lateral parabrachial nucleus (LPBN) in the behavioral control of body fluid homeostasis by determining the effect of bilateral injections of the CRH receptor antagonist, alpha-helical corticotropin-releasing factor (CRF)(9-41), and the CRH receptor agonist, CRH, on sodium chloride (salt appetite) and water (thirst) intake. Groups of adult, male Sprague-Dawley rats had stainless-steel cannulas implanted bilaterally into the LPBN and were sodium depleted or water deprived. Bilateral injections of alpha-helical CRF(9-41) into the LPBN significantly potentiated water and salt intake in the sodium-depleted rats when access to fluids was restored. Bilateral injections of alpha-helical CRF(9-41) into the LPBN (1.0 microg) also increased sodium appetite in water-deprived rats. Conversely, in sodium-depleted animals, bilateral injections of CRH inhibited sodium chloride intake. These results suggest that there is an endogenous CRH inhibitory mechanism operating in the LPBN to modulate the intake of sodium (salt appetite). This mechanism may contribute to the behavioral control of restoration of body fluid homeostasis in sodium-deficient states. PMID- 16357102 TI - Adrenal splanchnic innervation contributes to the diurnal rhythm of plasma corticosterone in rats by modulating adrenal sensitivity to ACTH. AB - Activity of the hypothalamic-pituitary-adrenal axis is characterized by a diurnal rhythm with an AM nadir and PM peak. Splanchnic nerve transection disrupts the diurnal rhythm in plasma corticosterone; however, there is a controversy as to whether the nerve-mediated effect is 1) via inhibition in the AM vs. excitation in the PM, or 2) involves changes in adrenal sensitivity to ACTH. The present studies were designed to address these issues. Adult male rats were anesthetized and underwent bilateral transection of the thoracic splanchnic nerve or sham transection. One week after surgery, rats were killed in the AM or PM with collection of nonstress plasma for measurement of corticosterone and ACTH. Plasma corticosterone was increased in the PM relative to the AM; however, plasma corticosterone in the PM was attenuated by splanchnic nerve transection, without affecting plasma ACTH. This decrease in PM plasma corticosterone after nerve transection was 1) associated with decreased adrenal responsivity to ACTH, 2) associated with decreased adrenal cAMP content, 3) prevented by adrenal demedullation, and 4) not affected by removal of adrenal capsaicin-sensitive afferent fibers. Repeated serial blood sampling from individual rats confirmed the excitatory effect of splanchnic innervation in the PM. These results support the hypothesis that the adrenal splanchnic innervation modulates the diurnal rhythm in plasma corticosterone by increasing adrenal responsivity to ACTH and augmenting steroidogenesis in the PM and suggest that alterations in adrenal corticosterone secretion obscured by pulsatile secretion are more clearly revealed with repeated serial blood sampling. PMID- 16357103 TI - An essential role of the CAAT/enhancer binding protein-alpha in the vitamin D induced expression of the human steroid/bile acid-sulfotransferase (SULT2A1). AB - The vitamin D receptor (VDR) regulates steroid and drug metabolism by inducing the genes encoding phase I and phase II enzymes. SULT2A1 is a liver- and intestine-expressed sulfo-conjugating enzyme that converts the alcohol-OH of neutral steroids, bile acids, and drugs to water-soluble sulfated metabolites. 1alpha,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] induces SULT2A1 gene transcription after the recruitment of VDR to the vitamin D-responsive chromatin region of SULT2A1. A composite element in human SULT2A1 directs the 1,25-(OH)2D3-mediated induction of natural and heterologous promoters. This element combines a VDR/retinoid X receptor-alpha-binding site [vitamin D response element (VDRE)], which is an imperfect inverted repeat 2 of AGCTCA, and a CAAT/enhancer binding protein (C/EBP)-binding site located 9 bp downstream to VDRE. The binding sites were identified by EMSA, antibody supershift, and deoxyribonuclease I footprinting. C/EBP-alpha at the composite element plays an essential role in the VDR regulation of SULT2A1, because 1) induction was lost for promoters with inactivating mutations at the VDRE or C/EBP element; 2) SULT2A1 induction by 1,25 (OH)2D3 in C/EBP-alpha-deficient cells required the expression of cotransfected C/EBP-alpha; and 3) C/EBP-beta did not substitute for C/EBP-alpha in this regulation. VDR and C/EBP-alpha were recruited concurrently to the composite element along with the coactivators p300, steroid receptor coactivator 1 (SRC-1), and SRC-2, but not SRC-3. VDR and C/EBP-alpha associated endogenously as a DNA dependent, coimmunoprecipitable complex, which was detected at a markedly higher level in 1,25-(OH)2D3-treated cells. These results provide the first example of the essential role of the interaction in cis between C/EBP-alpha and VDR in directing 1,25-(OH)2D3-induced expression of a VDR target gene. PMID- 16357104 TI - Identification of a splicing enhancer in MLH1 using COMPARE, a new assay for determination of relative RNA splicing efficiencies. AB - Exonic splicing enhancers (ESEs) are sequences that facilitate recognition of splice sites and prevent exon-skipping. Because ESEs are often embedded within protein-coding sequences, alterations in them can also often be interpreted as nonsense, missense or silent mutations. To correctly interpret exonic mutations and their roles in diseases, it is important to develop strategies that identify ESE mutations. Potential ESEs can be found computationally in many exons but it has proven difficult to predict whether a given mutation will have effects on splicing based on sequence alone. Here, we describe a flexible in vitro method that can be used to functionally compare the effects of multiple sequence variants on ESE activity in a single in vitro splicing reaction. We have applied this method in parallel with conventional splicing assays to test for a splicing enhancer in exon 17 of the human MLH1 gene. Point mutations associated with hereditary non-polyposis colorectal cancer (HNPCC) have previously been found to correlate with exon-skipping in both lymphocytes and tumors from patients. We show that sequences from this exon can replace an ESE from the mouse IgM gene to support RNA splicing in HeLa nuclear extracts. ESE activity was reduced by HNPCC point mutations in codon 659, indicating that their primary effect is on splicing. Surprisingly, the strongest enhancer function mapped to a different region of the exon upstream of this codon. Together, our results indicate that HNPCC point mutations in codon 659 affect an auxillary element that augments the enhancer function to ensure exon inclusion. PMID- 16357105 TI - Xcat, a novel mouse model for Nance-Horan syndrome inhibits expression of the cytoplasmic-targeted Nhs1 isoform. AB - Nance-Horan syndrome (NHS) is an X-linked disorder characterized by congenital cataracts, dental anomalies, dysmorphic features and mental retardation. A recent report suggests that the novel gene NHS1 is involved in this disorder due to the presence of point mutations in NHS patients. A possible mouse model for NHS, Xcat, was mapped to a 2.11 Mb interval on the X-chromosome. Sequence and FISH analysis of the X-chromosome region containing the Xcat mutation reveal a large insertion between exons 1 and 2 of the mouse Nhs1 gene. The insertion inhibits the expression of the Nhs1 isoform containing exon 1 and results in exclusive expression of the alternative isoform containing exon 1A. Quantitative RT-PCR of Xcat cDNA shows reduced levels of Nhs1 transcripts. The Nhs1 protein is strongly expressed within the cytoplasm of elongating lens fiber cells from wild-type neonate lens, but is significantly reduced within the Xcat lens. Transient transfection studies of CHO cells with Nhs1-GFP fusion proteins were done to determine whether the amino acids encoded by exon 1 were critical for protein localization. We found the presence of Nhs1 exon 1 critical for localization of the fusion protein to the cytoplasm, whereas fusion proteins lacking Nhs1 exon 1 are predominantly nuclear. These results indicate that the first exon of Nhs1 contains crucial information required for the proper expression and localization of Nhs1 protein. Inhibition of expression of the exon 1 containing isoform results in the abnormal phenotype of Xcat. PMID- 16357106 TI - Cross-species analyses implicate Lipin 1 involvement in human glucose metabolism. AB - Recent studies in the mouse have demonstrated that variations in lipin expression levels in adipose tissue have marked effects on adipose tissue mass and insulin sensitivity. In the mouse, lipin deficiency prevents normal adipose tissue development, resulting in lipodystrophy and insulin resistance, whereas excess lipin levels promote fat accumulation and insulin sensitivity. Here, we investigated the effects of genetic variation in lipin levels on glucose homeostasis across species by analyzing lipin transcript levels in human and mouse adipose tissues. A strong negative correlation was observed between lipin mRNA levels and fasting glucose and insulin levels, as well as an indicator of insulin resistance (HOMA-IR), in both mice and humans. We subsequently analyzed the allelic diversity of the LPIN1 gene in dyslipidemic Finnish families, as well as in a case-control sample of obese (n = 477) and lean (n = 821) individuals. Alleles were defined by genotyping seven single nucleotide polymorphisms (SNPs) of the critical DNA region over the LPIN1 gene. Intragenic SNPs and corresponding allelic haplotypes exhibited associations with serum insulin levels and body mass index (P = 0.002-0.04). Both the expression levels in adipose tissue across species and genetic data in human study samples highlight the importance of lipin in glucose homeostasis and imply that allelic variants of this gene have significance in human metabolic traits. PMID- 16357107 TI - Hemostatic factors, inflammatory markers, and progressive peripheral atherosclerosis: the Edinburgh Artery Study. AB - The interplay between inflammatory and hemostatic mechanisms may play a crucial role in the development and progression of atherosclerosis. The authors evaluated the separate and joint associations of hemostatic and inflammatory variables on peripheral atherosclerotic progression in the Edinburgh Artery Study, a population cohort study of 1,592 men and women aged 55-74 years that started in 1987. Levels of fibrinogen, fibrin D-dimer, von Willebrand factor, tissue plasminogen activator antigen, factor VII, prothrombin fragment 1 + 2, urinary fibrinopeptide A, C-reactive protein, and interleukin-6 were measured at baseline. Arm and ankle blood pressures were measured, and atherosclerotic progression was assessed by computing ankle brachial index (ABI) at baseline (1,582 participants) and after 12 years of follow-up (813 participants). Fibrinogen (p = 0.05) and D-dimer (p < or = 0.05) were significantly associated with ABI change independently of baseline ABI and cardiovascular disease risk factors. However, these associations were no longer significant when analyses were adjusted for either C-reactive protein or interleukin-6. Moreover, subjects with higher levels of both D-dimer and interleukin-6 at baseline had the greatest ABI decline. In conclusion, fibrinogen and D-dimer, but not other hemostatic factors, were associated with progressive peripheral atherosclerosis. Since D dimer and fibrinogen are acute phase reactants, these data support the hypothesis that inflammation is more related to atherosclerosis than is hypercoagulation. PMID- 16357108 TI - Exposure to loud noise and risk of acoustic neuroma. AB - Exposure to occupational loud noise has been previously identified as a possible risk factor for acoustic neuroma in only one relatively small (n = 86 cases) case control study of men. The goal of the present study was to further examine the role of loud noise in acoustic neuroma etiology. In their population-based case control study of both sexes conducted from 1999 to 2002 in Sweden, the authors compared reports on type and duration of occupational and nonoccupational loud noise exposure of 146 acoustic neuroma cases and 564 controls. Controls were randomly selected from the study base and were frequency matched on age, sex, and residential area. The authors found that individuals reporting loud noise exposure from any source were at increased risk for acoustic neuroma (odds ratio (OR) = 1.55, 95% confidence interval (CI): 1.04, 2.30). Exposure to loud noise from machines, power tools, and/or construction increased the risk for acoustic neuroma (OR = 1.79, 95% CI: 1.11, 2.89), as did exposure to loud music (OR = 2.25, 95% CI: 1.20, 4.23). The odds ratio for a latency period of 13 or more years since the first loud noise exposure from any source was 2.12 (95% CI: 1.40, 3.20). The findings of an increased risk of acoustic neuroma with loud noise exposure support previous research. PMID- 16357109 TI - Palmtop-assisted self-interviewing for the collection of sensitive behavioral data: randomized trial with drug use urine testing. AB - Palmtop-assisted self-interviewing (PASI) may provide a cheaper and more mobile alternative to audio-computer-assisted self-interviewing (ACASI) for collecting sensitive behavioral data. To evaluate PASI, in late 2002 the authors enrolled 1,283 Thai students aged 15-21 years in a randomized trial. Data collection used PASI, ACASI, self-administered questionnaire, and face-to-face interview in combination with drug-use urine testing. By use of reported levels of behaviors and agreement between self-reports of smoking and urine test results, PASI and ACASI (alpha = 0.05) were compared for noninferiority, and PASI and interview were compared for superiority (alpha = 0.05). Noninferiority of PASI was demonstrated by use of self-reports of the most sensitive areas of sexual behavior (e.g., oral sex, sexual intercourse, commercial sex, history of genital ulcers, pregnancy), as well as self-reports of less sensitive behaviors (e.g., alcohol use, dietary behaviors, symptoms of depression). Data generally showed noninferiority of PASI, ACASI, and self-administered questionnaires when compared with each other and superiority of PASI, ACASI, and self-administered questionnaires when compared with interviews. PASI agreements between self reports of tobacco smoking and presence of nicotine metabolites in urine were noninferior to ACASI and superior to interviews. The establishment of PASI noninferiority and superiority using behavioral and biologic measures suggests that PASI is a scientifically acceptable alternative for collecting sensitive behavioral data. PMID- 16357110 TI - Genetic variation in the sodium-dependent vitamin C transporters, SLC23A1, and SLC23A2 and risk for preterm delivery. AB - Vitamin C has been the focus of epidemiologic investigation in preterm delivery (<37 weeks' gestation), which is a leading cause of neonatal mortality and birth related morbidity. There are two sodium-dependent membrane transporters encoded by SLC23A1 and SLC23A2, which have key roles in human vitamin C metabolism and which control dietary uptake, reabsorption, and tissue distribution of vitamin C. Using maternal DNA, the authors evaluated common single-nucleotide polymorphisms (SNPs) in SLC23A1 and SLC23A2 in a nested case-control analysis of the Pregnancy, Infection, and Nutrition Study (1995-2000) cohort. Of the associations observed for both haplotypes in SLC23A1 and individual SNPs in SLC23A2, the most robust finding is with an intron 2 variant in SLC23A2. Heterozygotes and homozygotes for this variant had a 1.7-fold (95% confidence interval: 0.9, 3.3) and a 2.7-fold (95% confidence interval: 1.2, 6.3) elevation in the risk of spontaneous preterm birth, respectively. Semi-Bayesian hierarchical regression analysis, which simultaneously adjusted for multiple SNPs within the same gene, gave comparable results. The authors' findings link genetic variants in the vitamin C transporters to spontaneous preterm birth, which may explain previous dietary associations. If the findings from this study are confirmed, they may serve as the foundation for genetic risk assessment of nutritional pathways in preterm birth. PMID- 16357111 TI - Nut and seed consumption and inflammatory markers in the multi-ethnic study of atherosclerosis. AB - Nuts and seeds are rich in unsaturated fat and other nutrients that may reduce inflammation. Frequent nut consumption is associated with lower risk of cardiovascular disease and type 2 diabetes. The authors examined associations between nut and seed consumption and C-reactive protein, interleukin-6, and fibrinogen in the Multi-Ethnic Study of Atherosclerosis. This 2000 cross sectional analysis included 6,080 US participants aged 45-84 years with adequate information on diet and biomarkers. Nut and seed consumption was categorized as never/rare, less than once/week, 1-4 times/week, and five or more times/week. After adjustment for age, gender, race/ethnicity, site, education, income, smoking, physical activity, use of fish oil supplements, and other dietary factors, mean biomarker levels in categories of increasing consumption were as follows: C-reactive protein-1.98, 1.97, 1.80, and 1.72 mg/liter; interleukin-6 1.25, 1.24, 1.21, and 1.15 pg/ml; and fibrinogen-343, 338, 338, and 331 mg/dl (all p's for trend < 0.01). Further adjustment for hypertension, diabetes, medication use, and lipid levels yielded similar results. Additional adjustment for body mass index moderately attenuated the magnitude of the associations, yielding borderline statistical significance. Associations of nut and seed consumption with these biomarkers were not modified by body mass index, waist:hip ratio, or race/ethnicity. Frequent nut and seed consumption was associated with lower levels of inflammatory markers, which may partially explain the inverse association of nut consumption with cardiovascular disease and diabetes risk. PMID- 16357112 TI - Effect of fetal sex on airway lability in pregnant women with asthma. AB - The authors investigated the association between sex of the fetus and maternal airway lability among pregnant women with asthma. Data were prospectively collected among 702 pregnant women with a diagnosis of asthma who were recruited in southern New England between 1997 and 2000 and followed through pregnancy. Peak expiratory flow lability, defined as percent daily maximum minus the minimum divided by the mean, was assessed at enrollment and at 21, 29, and 37 weeks' gestation. There was a -9.9 percent (95 percent confidence interval: -19.4, -0.4) difference in airway lability observed between women carrying female fetuses and those carrying male fetuses. This difference persisted throughout pregnancy. Among pregnant asthmatic women, carrying a female fetus is associated with worse maternal asthma, as assessed by greater airway lability, than is carrying a male fetus. PMID- 16357113 TI - Malaria mapping using transmission models: application to survey data from Mali. AB - Geographic mapping of the distribution of malaria is complicated by the limitations of the available data. The most widely available data are from prevalence surveys, but these surveys are generally carried out at arbitrary locations and include nonstandardized and overlapping age groups. To achieve comparability between different surveys, the authors propose the use of transmission models, particularly the Garki model, to convert heterogeneous age prevalence data to a common scale of estimated entomological inoculation rates, vectorial capacity, or force of infection. They apply this approach to the analysis of survey data from Mali, collected in 1965-1998, extracted from the Mapping Malaria Risk in Africa database. They use Bayesian geostatistical models to produce smooth maps of estimates of the entomological inoculation rates obtained from the Garki model, allowing for the effect of environmental covariates. Again using the Garki model, they convert kriged entomological inoculation rates values to age-specific malaria prevalence. The approach makes more efficient use of the available data than do previous malaria mapping methods, and it produces highly plausible maps of malaria distribution. PMID- 16357114 TI - Acute pain management: scientific evidence revisited. PMID- 16357115 TI - Deadspace: invasive or not? PMID- 16357116 TI - Org 25969 (sugammadex), a selective relaxant binding agent for antagonism of prolonged rocuronium-induced neuromuscular block. AB - BACKGROUND: Org 25969 is a cyclodextrin compound designed to reverse a rocuronium induced neuromuscular block. The aim of this study was to explore the efficacy, dose-response relation and safety of Org 25969 for reversal of a prolonged rocuronium-induced neuromuscular block. METHODS: Thirty anaesthetized adult patients received rocuronium 0.6 mg kg(-1) as an initial dose followed by increments to maintain a deep block at a level of <10 PTCs (post-tetanic counts) recorded every 6 min. Neuromuscular monitoring was carried out using accelerometry, in a train-of-four (TOF) mode using TOF-WatchSX. At recovery of T2, following at least 2 h of neuromuscular block, patients received their randomly assigned dose of 0.5, 1.0, 2.0, 4.0 or 6.0 mg kg(-1) of Org 25969. Anaesthesia and neuromuscular monitoring were continued for a minimum period of 30 min after Org 25969 administration. The main end-point of the study was the time to achieve a sustained recovery of TOF ratio to 0.9. Patients were followed up for 7 days after anaesthesia. RESULTS: The results showed a dose-related decrease in the average time taken to attain a TOF ratio of 0.9 from 6:49 (min:s) with the 0.5 mg kg(-1) dose to 1:22 with the 4.0 mg kg(-1) dose. Weighted non linear regression analysis showed the fastest achievable time to TOF ratio of 0.9 to be 1:35. Org 25969 produced no major adverse effects. CONCLUSION: Org 25969 effectively reversed a deep and prolonged neuromuscular block induced by rocuronium. The effective reversal dose appears to be 2-4 mg kg(-1). PMID- 16357117 TI - Use of a stimulating catheter for femoral nerve block. PMID- 16357118 TI - Hyperinsulinaemic normoglycaemic clamp in coronary artery surgery. PMID- 16357119 TI - S100B in Guillain-Barre syndrome. PMID- 16357120 TI - Laryngospasm during subarachnoid block. PMID- 16357122 TI - It is time for a Human Epigenome Project. PMID- 16357124 TI - Loss of imprinting of IGF2: a common epigenetic modifier of intestinal tumor risk. AB - Epigenetic alterations in cancer occur at least as commonly as genetic mutations, but epigenetic alterations could occur secondarily to the tumor process itself. To establish a causal role of epigenetic changes, investigators have turned to genetically engineered mouse models. Here, we review a recent study showing that a mouse model of loss of imprinting (LOI) of the insulin-like growth factor II gene (Igf2), which shows aberrant activation of the normally silent maternal allele, modifies the risk of intestinal neoplasia caused by mutations of the adenomatous polyposis coli (Apc) gene. This increased risk corresponds to the apparent increased risk of colorectal cancer in patients with LOI of IGF2. The model suggests that preexisting epigenetic alterations in normal cells increase tumor risk by expanding the target cell population and/or modulating the effect of subsequent genetic alterations on these cells, providing a novel idea for cancer risk management. PMID- 16357123 TI - Anoikis and survival connections in the tumor microenvironment: is there a role in prostate cancer metastasis? AB - Overcoming the androgen independence of prostate tumors is considered the most critical therapeutic end point for improving survival in patients with metastatic prostate cancer. Normal epithelial and endothelial cells can undergo apoptosis when detached from the extracellular matrix (ECM), via the anoikis phenomenon. In contrast, tumor cells upon detachment from the ECM are capable of evading anoikis and metastasizing to different distant organs. Is the biological repertoire of the epithelial and endothelial cells sufficient to account for the events associated with the process of anoikis during prostate cancer metastasis? Although there is no clear answer to this question, what has become increasingly evident from the existing evidence is that molecules that induce anoikis in tumor epithelial and endothelial cells provide exciting new leads into effective therapeutic targeting as well as markers of prostate cancer progression and prediction of therapeutic resistance. This review analyzes recent findings on anoikis regulators and discusses the relevance of this unique apoptosis mode in the development of metastatic prostate cancer and identification of molecular signatures for treatment of advanced disease. PMID- 16357125 TI - A blueprint for a Human Epigenome Project: the AACR Human Epigenome Workshop. AB - Epigenetic processes control the packaging and function of the human genome and contribute to normal and pathologic states, including cancer. The time is ripe to undertake an international effort to identify all the chemical changes and relationships between chromatin constituents that provide function to the genetic code. A timely workshop of leading experts, convened by the American Association for Cancer Research (AACR), confirmed that the technology is at hand to begin defining human epigenomes at high resolution. PMID- 16357126 TI - Modeling human cancer: report on the Eighth Beatson International Cancer Conference. AB - One of the major challenges facing cancer researchers is the necessity to adequately model the human disease. This need is driven by goals, including validating/determining the contribution of a gene to cancer, establishing systems for drug discovery and validation, and discovering additional factors that modify the oncogenic process. Recent advances in cancer modeling were discussed at the Eighth Beatson International Cancer Conference on June 19 to 22, 2005 in Glasgow, United Kingdom. The conference was attended by 250 scientists with >130 oral and poster presentations. The research presented made use of systems ranging from two dimensional and three-dimensional cell culture to nonmammalian and mammalian in vivo models. In addition to basic research on the fundamental processes involved in cancer, novel developments in cancer treatment were presented. PMID- 16357127 TI - Seventeenth Annual Pezcoller Symposium: molecular understanding of solid tumors. PMID- 16357128 TI - Cyclophosphamide allows for in vivo dose reduction of a potent oncolytic virus. AB - The success of cancer virotherapy depends on its efficacy versus toxicity profile in human clinical trials. Progress towards clinical trials can be hampered by the relatively elevated doses of oncolytic viruses administered in animal models to achieve an anticancer effect and by the even higher doses required in humans to approximate an animal bioequivalent dose. Such elevated doses of injected viral proteins may also lead to undesirable toxicities and are also very difficult to produce in a biotechnological setting. We report that a relatively potent herpes simplex virus type 1 oncolytic virus (rQNestin34.5) produces 45% survivors at a dose of 3 x 10(4) plaque-forming units (pfu) in a 9-day-old mouse model of human glioma. Unlike our previous findings with less potent oncolytic viruses, though, the preadministration of cyclophosphamide did not enhance this survival or affect oncolytic virus tumor distribution and tumor volume. However, when oncolytic virus doses were reduced (3 x 10(3) and 3 x 10(2) pfu), cyclophosphamide significantly enhanced both animal survival and oncolytic virus tumor distribution and also reduced tumor volumes. These findings thus show that cyclophosphamide allows for dose reduction of doses of a relatively potent oncolytic virus, a finding with implications for the development of clinical trials. PMID- 16357129 TI - Identification of GATA3 as a breast cancer prognostic marker by global gene expression meta-analysis. AB - GATA binding protein 3 (GATA3) is a transcriptional activator highly expressed by the luminal epithelial cells in the breast. Here we did a meta-analysis of the available breast cancer cDNA data sets on a cohort of 305 patients and found that GATA3 was one of the top genes with low expression in invasive carcinomas with poor clinical outcome. To validate its prognostic utility, we did a tissue microarray analysis on a cohort of 139 consecutive invasive carcinomas (n = 417 tissue samples) immunostained with a monoclonal antibody against GATA3. Low GATA3 expression was associated with higher histologic grade (P < 0.001), positive nodes (P = 0.002), larger tumor size (P = 0.03), negative estrogen receptor and progesterone receptor (P < 0.001 for both), and HER2-neu overexpression (P = 0.03). Patients whose tumors expressed low GATA3 had significantly shorter overall and disease-free survival when compared with those whose tumors had high GATA3 levels. The hazard ratio of metastasis or recurrence according to the GATA3 status was 0.31 (95% confidence interval, 0.13-0.74; P = 0.009). Cox multivariate analysis showed that GATA3 had independent prognostic significance above and beyond conventional variables. Our data suggest that immunohistochemical analysis of GATA3 may be the basis for a new clinically applicable test to predict tumor recurrence early in the progression of breast cancer. PMID- 16357130 TI - TRAIL receptor-selective mutants signal to apoptosis via TRAIL-R1 in primary lymphoid malignancies. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its agonistic antibodies, which are currently in early clinical trials for treating various malignancies, induce apoptosis through triggering of either TRAIL-R1 or TRAIL-R2. Based on studies using agonistic monoclonal antibodies, we recently proposed that primary chronic lymphocytic leukemic cells seem to signal apoptosis primarily through TRAIL-R1. We have now synthesized mutant forms of TRAIL specific for TRAIL-R1 or TRAIL-R2. The selectivity of these mutants to induce apoptosis in cell lines is due to selective binding to their cognate receptors resulting in apoptosis via formation of a death-inducing signaling complex. Using these mutants, we now unequivocally show that primary cells from patients with chronic lymphocytic leukemia and mantle cell lymphoma signal to apoptosis almost exclusively through TRAIL-R1. Thus, no significant therapeutic benefit can be anticipated from treating such patients with agents currently in clinical trials that signal predominantly through TRAIL-R2, such as HGS-ETR2 or Apo2L/TRAIL. Our study highlights the necessity to determine whether primary cells from a particular tumor signal via TRAIL-R1 or TRAIL-R2. Such information will provide a rational approach to optimize TRAIL therapy. PMID- 16357131 TI - Organ-specific increase in mutation accumulation and apoptosis rate in CuZn superoxide dismutase-deficient mice. AB - Reactive oxygen species have been implicated as a cause of cancer and aging in mammals. Mice deficient for the antioxidant enzyme CuZn-superoxide dismutase (Sod1) have a decreased life span and an elevated incidence of liver cancer. To test the hypothesis that the cancer-prone phenotype in such mice is due to accelerated spontaneous mutation accumulation, we crossed these mutants with mice harboring a neutral lacZ mutation reporter gene. At 2 months of age, the lacZ mutation frequency in the liver of the hybrid animals was already twice as high as in littermate controls of the same age. This difference in mutation frequency increased to >3-fold at 6 months of age, after which it did not increase any further. Characterization of the mutation spectra in liver of the Sod1-null mice indicated mainly GC-to-TA transversions and GC-to-AT transitions, signature mutations of oxidative stress. The accelerated mutation accumulation in liver was accompanied by an increased frequency of apoptotic cells, as indicated by an increase in both terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling- and caspase 3-stained cells at 6 and 12 months of age. In kidney, an elevated mutation frequency above controls of approximately 2.5-fold was found not earlier than at 6 months. No increased mutation accumulation was observed in brain or spleen. These results support the hypothesis, that oxidative stress is an important causal factor of cancer in mammals. PMID- 16357132 TI - Loss of phosphatase and tensin homologue increases transforming growth factor beta-mediated invasion with enhanced SMAD3 transcriptional activity. AB - In normal epithelial tissues, the multifunctional cytokine transforming growth factor-beta (TGF-beta) acts as a tumor suppressor through growth inhibition and induction of differentiation whereas in advanced cancers, TGF-beta promotes tumor progression through induction of tumor invasion, neoangiogenesis, and immunosuppression. The molecular mechanisms through which TGF-beta shifts from a tumor suppressor to a tumor enhancer are poorly understood. We now show a role for the tumor suppressor phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in repressing the protumorigenic effects of TGF-beta. The TGF-beta effector SMAD3 inducibly interacts with PTEN on TGF-beta treatment under endogenous conditions. RNA interference (RNAi) suppression of PTEN expression enhances SMAD3 transcriptional activity and TGF-beta-mediated induction of SMAD3 target genes whereas reconstitution of PTEN in a null cancer cell line represses the expression of TGF-beta-regulated target genes. Targeting PTEN expression through RNAi in a PTEN wild-type cell line increases TGF-beta-mediated invasion but does not affect TGF-beta-mediated growth inhibition. Reconstitution of PTEN expression in a PTEN-null cell line blocks TGF-beta-induced invasion but does not modulate TGF-beta-mediated growth regulation. These effects are distinct from Akt and Forkhead family members that also interact with SMAD3 to regulate apoptosis or proliferation, respectively. Pharmacologic inhibitors targeting TGF-beta receptors and phosphatidylinositol 3-kinase signaling downstream from PTEN cooperate to block TGF-beta-mediated invasion. Thus, the loss of PTEN expression in human cancers may contribute to a role for TGF-beta as a tumor enhancer with specific effects on cellular motility and invasion. PMID- 16357133 TI - Akt phosphorylates and regulates Pdcd4 tumor suppressor protein. AB - Programmed cell death 4 (Pdcd4) is a tumor suppressor protein that interacts with eukaryotic initiation factor 4A and inhibits protein synthesis. Pdcd4 also suppresses the transactivation of activator protein-1 (AP-1)-responsive promoters by c-Jun. The Akt (protein kinase B) serine/threonine kinase is a key mediator of phosphoinositide 3-kinase pathway involved in the regulation of cell proliferation, survival, and growth. Because Pdcd4 has two putative Akt phosphorylation sites at Ser(67) and Ser(457), we investigated whether Akt phosphorylates and regulates Pdcd4. Our results show that Akt specifically phosphorylates Ser(67) and Ser(457) residues of Pdcd4 in vitro and in vivo. We further show that phosphorylation of Pdcd4 by Akt causes nuclear translocation of Pdcd4. Using luciferase assay, we show that phosphorylation of Pdcd4 by Akt also causes a significant decrease of the ability of Pdcd4 to interfere with the transactivation of AP-1-responsive promoter by c-Jun. PMID- 16357134 TI - Aromatase inhibitors in human lung cancer therapy. AB - Lung cancer is the most common cancer in the world. It is a highly lethal disease in women and men, and new treatments are urgently needed. Previous studies implicated a role of estrogens and estrogen receptors in lung cancer progression, and this steroidal growth-stimulatory pathway may be promoted by tumor expression and activity of aromatase, an estrogen synthase. We found expression of aromatase transcripts and protein in human non-small cell lung cancer (NSCLC) cells using reverse transcription-PCR and Western immunoblots, respectively. Aromatase staining by immunohistochemistry was detected in 86% of archival NSCLC tumor specimens from the clinic. Further, biological activity of aromatase was determined in NSCLC tumors using radiolabeled substrate assays as well as measure of estradiol product using ELISA. Significant activity of aromatase occurred in human NSCLC tumors, with enhanced levels in tumor cells compared with that in nearby normal cells. Lung tumor aromatase activity was inhibited by anastrozole, an aromatase inhibitor, and treatment of tumor cells in vitro with anastrozole led to significant suppression of tumor cell growth. Similarly, among ovariectomized nude mice with A549 lung tumor xenografts, administration of anastrozole by p.o. gavage for 21 days elicited pronounced inhibition of tumor growth in vivo. These findings show that aromatase is present and biologically active in human NSCLCs and that tumor growth can be down-regulated by specific inhibition of aromatase. This work may lead to development of new treatment options for patients afflicted with NSCLC. PMID- 16357135 TI - Checkpoint abrogation in G2 compromises repair of chromosomal breaks in ataxia telangiectasia cells. AB - Checkpoint abrogation in G(2) compromises repair of DNA double-strand breaks (DSB) and confers enhanced G(2) chromosomal radiosensitivity in ataxia telangiectasia (AT) cells. To directly test this hypothesis, we combined calyculin A-induced premature chromosome condensation with conventional cytogenetics to evaluate chromosome damage before and after the G(2) checkpoint in irradiated primary AT and normal human lymphocytes and their lymphoblastoid derivatives. Direct analysis of radiation damage in G(2) by premature chromosome condensation reveals practically indistinguishable levels of chromosomal breaks in AT and normal cells. Yet a 4-fold increase in metaphase chromosome damage is observed in AT cells as compared with normal cells which, in contrast to AT cells, exhibit a strong G(2) arrest manifest as an abrupt reduction in the mitotic index. Thus, an active checkpoint facilitates repair of chromosomal breaks in normal cells. Treatment with caffeine that abrogates the G(2) checkpoint without significantly affecting DSB rejoining increases metaphase chromosome damage of normal cells to the AT level but leaves unchanged interphase chromosome damage in G(2). Caffeine has no effect on any of these end points in AT cells. These observations represent the first direct evidence that the G(2) checkpoint facilitates repair of chromosome damage, presumably by supporting repair of DNA DSBs. Failure to arrest will lead to chromatin condensation and conversion of unrepaired DNA DSBs to chromosomal breaks during G(2)-to-M phase transition. PMID- 16357136 TI - Mutation of Lkb1 and p53 genes exert a cooperative effect on tumorigenesis. AB - Peutz-Jeghers syndrome (PJS) is a dominantly inherited disorder characterized by gastrointestinal hamartomatous polyps and mucocutaneous melanin pigmentation. Germ line mutations in LKB1 cause PJS. We have generated mice carrying an Lkb1 exon 2 to 8 deletion by gene targeting in embryonic stem cells. Heterozygotes develop gastric hamartomas that are histologically similar to those found in humans with PJS. LKB1 is also reportedly a mediator of p53-dependent apoptosis. To explore the potential combined effects of p53 and Lkb1 alterations on tumorigenesis, we carried out a series of matings with Lkb1(+/-) and p53 null mice to generate Lkb1(+/-)/p53(+/-) and Lkb1(+/-)/p53(-/-) mice. Similar to the Lkb1(+/-) mice, gastrointestinal hamartomas have also been detected in the mice with these two genotypes. The Lkb1(+/-)/p53(+/-) mice displayed a dramatically reduced life span and increased tumor incidence compared to the mice with either Lkb1 or p53 single gene knockout. The time to onset of polyposis in Lkb1(+/ )/p53(-/-) mice is approximately 2 months earlier than Lkb1(+/-)/p53(+/-) and Lkb1(+/-) mice, whereas the latter two show a similar time to onset which is at approximately 6 months of age. These results strongly suggested that mutations of p53 and Lkb1 gene cooperate in the acceleration of tumorigenesis. PMID- 16357137 TI - FBXO31 is the chromosome 16q24.3 senescence gene, a candidate breast tumor suppressor, and a component of an SCF complex. AB - A BAC located in the 16q24.3 breast cancer loss of heterozygosity region was previously shown to restore cellular senescence when transferred into breast tumor cell lines. We have shown that FBXO31, although located just distal to this BAC, can induce cellular senescence in the breast cancer cell line MCF-7 and is the likely candidate senescence gene. FBXO31 has properties consistent with a tumor suppressor, because ectopic expression of FBXO31 in two breast cancer cell lines inhibited colony growth on plastic and inhibited cell proliferation in the MCF-7 cell line. In addition, compared with the relative expression in normal breast, levels of FBXO31 were down-regulated in breast tumor cell lines and primary tumors. FBXO31 was cell cycle regulated in the breast cell lines MCF-10A and SKBR3 with maximal expression from late G(2) to early G(1) phase. Ectopic expression of FBXO31 in the breast cancer cell line MDA-MB-468 resulted in the accumulation of cells at the G(1) phase of the cell cycle. FBXO31 contains an F box domain and is associated with the proteins Skp1, Roc-1, and Cullin-1, suggesting that FBXO31 is a component of a SCF ubiquitination complex. We propose that FBXO31 functions as a tumor suppressor by generating SCF(FBXO31) complexes that target particular substrates, critical for the normal execution of the cell cycle, for ubiquitination and subsequent degradation. PMID- 16357138 TI - ANLN plays a critical role in human lung carcinogenesis through the activation of RHOA and by involvement in the phosphoinositide 3-kinase/AKT pathway. AB - Gene expression profile analysis of non-small cell lung cancers (NSCLC) and subsequent functional analyses revealed that human ANLN, a homologue of anillin, an actin-binding protein in Drosophila, was transactivated in lung cancer cells and seemed to play a significant role in pulmonary carcinogenesis. Induction of small interfering RNAs against ANLN in NSCLC cells suppressed its expression and resulted in growth suppression; moreover, treatment with small interfering RNA yielded cells with larger morphology and multiple nuclei, which subsequently died. On the other hand, induction of exogenous expression of ANLN enhanced the migrating ability of mammalian cells by interacting with RHOA, a small guanosine triphosphatase, and inducing actin stress fibers. Interestingly, inhibition of phosphoinositide 3-kinase/AKT activity in NSCLC cells decreased the stability of ANLN and caused a reduction of the nuclear ANLN level. Immunohistochemical staining of nuclear ANLN on lung cancer tissue microarrays was associated with the poor survival of NSCLC patients, indicating that this molecule might serve as a prognostic indicator. Our data imply that up-regulation of ANLN is a common feature of the carcinogenetic process in lung tissue, and suggests that selective suppression of ANLN could be a promising approach for developing a new strategy to treat lung cancers. PMID- 16357139 TI - A putative role for psoriasin in breast tumor progression. AB - Psoriasin (S100A7) was identifi;ed as a gene highly expressed in psoriatic keratinocytes and highly and more frequently expressed in ductal carcinoma in situ (DCIS) than in invasive breast carcinomas (IBC), suggesting a potential role in tumor progression. Psoriasin expression is associated with poor prognostic factors in both DCIS and IBC. Several putative functions have been proposed for psoriasin in various disease types, but none of these can fully explain its involvement in breast tumor progression. Here, we show that down-regulation of endogenous psoriasin expression via stable short hairpin RNAs in a human IBC cell line (MDA-MB-468) increases cell migration and invasion without influencing cell proliferation and survival in vitro but inhibits tumor growth in vivo. These seemingly paradoxical results are potentially explained by the dramatic up regulation and down-regulation of matrix metalloproteinase-13 and vascular endothelial growth factor (VEGF), respectively, observed in cells with decreased psoriasin levels compared with controls. Correlating with this, high psoriasin expression in human IBC is associated with increased angiogenesis and worse clinical outcome, and psoriasin mRNA levels are coordinately regulated with VEGF and other genes related to hypoxia and mitochondrial reactive oxygen species (ROS). Based on these results, we propose that psoriasin may play a role in breast tumor progression by promoting angiogenesis and enhancing the selection for cells that overcome its anti-invasive function. This hypothesis may explain why psoriasin expression is highest in high-grade and/or estrogen receptor negative tumors, as these are associated with increased hypoxia and ROS, a setting in which the angiogenic effects of psoriasin are most important. PMID- 16357140 TI - Gene expression profiles associated with treatment response in oligodendrogliomas. AB - Oligodendrogliomas are a specific subtype of brain tumor of which the majority responds favorably to chemotherapy. In this study, we made use of expression profiling to identify chemosensitive oligodendroglial tumors. Correlation of expression profiles to loss of heterozygosity on 1p and 19q, common chromosomal aberrations associated with response to treatment, identified 376, 64, and 60 differentially expressed probe sets associated with loss of 1p, 19q or 1p, and 19q, respectively. Correlation of expression profiles to the tumors' response to treatment identified 16 differentially expressed probe sets. Because transcripts associated with chemotherapeutic response were identified independent of common chromosomal aberrations, expression profiling may be used as an alternative approach to the tumors' 1p status to identify chemosensitive oligodendroglial tumors. Finally, we correlated expression profiles to survival of the patient after diagnosis and identified 103 differentially expressed probe sets. The observation that many genes are differentially expressed between long and short survivors indicates that the genetic background of the tumor is an important factor in determining the prognosis of the patient. Furthermore, these transcripts can help identify patient subgroups that are associated with favorable prognosis. Our study is the first to correlate gene expression with chromosomal aberrations and clinical performance (response to treatment and survival) in oligodendrogliomas. The differentially expressed transcripts can help identify patient subgroups with good prognosis and those that will benefit from chemotherapeutic treatments. PMID- 16357141 TI - Mutually exclusive cyclin-dependent kinase 4/cyclin D1 and cyclin-dependent kinase 6/cyclin D2 pairing inactivates retinoblastoma protein and promotes cell cycle dysregulation in multiple myeloma. AB - Multiple myeloma, the second most common hematopoietic cancer, ultimately becomes refractory to treatment when self-renewing multiple myeloma cells begin unrestrained proliferation by unknown mechanisms. Here, we show that one, but not more than one, of the three early G(1) D cyclins is elevated in each case of multiple myeloma. Cyclin D1 or D3 expression does not vary in the clinical course, but that alone is insufficient to promote cell cycle progression unless cyclin-dependent kinase 4 (cdk4) is also elevated, in the absence of cdk6, to phosphorylate the retinoblastoma protein (Rb). By contrast, cyclin D2 and cdk6 are coordinately increased, thereby overriding the inhibition by cdk inhibitors p18(INK4c) and p27(Kip1) and phosphorylating Rb in conjunction with the existing cdk4. Thus, cyclin D1 pairs exclusively with cdk4 and cdk6 pairs only with cyclin D2, although cyclin D2 can also pair with cdk4 in multiple myeloma cells. The basis for this novel and specific cdk/D cyclin pairing lies in differential transcriptional activation. In addition, cyclin D1- or cyclin D3-expressing multiple myeloma cells are uniformly distributed in the bone marrow, whereas cdk6 specific phosphorylation of Rb occurs in discrete foci of bone marrow multiple myeloma cells before proliferation early in the clinical course and is then heightened with proliferation and disease progression. Mutually exclusive cdk4/cyclin D1 and cdk6/cyclin D2 pairing, therefore, is likely to be a critical determinant for cell cycle reentry and progression and may play a pivotal role in the expansion of self-renewing multiple myeloma cells. PMID- 16357142 TI - Negative regulation of TSC1-TSC2 by mammalian D-type cyclins. AB - The metazoan cell cycle is driven by the timely and composite activities of cyclin-dependent kinases (CDKs). Among these, cyclin D- and cyclin E-dependent kinases phosphorylate the pRb family proteins during G(1) phase of the cell cycle and thereby advance cells beyond the restriction point. Increasing evidence suggests that cyclin D-dependent kinases might affect events other than Rb pathway-mediated entry into S phase, such as accumulation of cell mass. However, little is known about cyclin D activity toward Rb-independent pathway(s) or non pRb substrates. In this article, we show that the tumor suppressor TSC2 is a cyclin D binding protein. Coexpression of cyclin D1-CDK4/6 in cultured cells leads to increased phosphorylation and decreased detection of both TSC2 and TSC1, and promotes the phosphorylation of the mTOR substrates, 4E-BP1 and S6K1, two key effectors of cell growth that are negatively regulated by the TSC1-TSC2 complex. At the cellular level, ectopic expression of cyclin D1 restores the cell size decrease caused by TSC1-TSC2 expression. Intriguingly, down-regulation of TSC proteins was also observed by the expression of a mutant cyclin D1 that is unable to bind to CDK4/6, or by the coexpression of cyclin D1 with either an INK4 inhibitor or with catalytically inactive CDK6, indicating that cyclin D may regulate TSC1-TSC2 independently of CDK4/6. Together, these observations suggest that mammalian D-type cyclins participate in cell growth control through negative regulation of TSC1-TSC2 function. PMID- 16357143 TI - CDC4 mutations occur in a subset of colorectal cancers but are not predicted to cause loss of function and are not associated with chromosomal instability. AB - CDC4/FBXW7 is part of a ubiquitin ligase complex which targets molecules such as cyclin E, c-myc, and c-jun for destruction. CDC4 mutations occur in several cancer types and are best described in colorectal tumors. Knockout of CDC4 in vitro in colorectal cancer cells causes changes suggestive of chromosomal instability (CIN). In p53(+/-) mice, radiation-induced lymphomas show deletion or mutation of one copy of CDC4 and knockdown of CDC4 leads to increased aneuploidy in mouse fibroblasts. We screened 244 colorectal tumors and 40 cell lines for CDC4 mutations and allelic loss. Six percent (18 of 284) of tumors, including near-diploid (CIN-) lesions, harbored CDC4 mutations and there was no association between mutation and CIN (polyploidy). The CDC4 mutation spectrum in colorectal tumors was heavily biased towards C:G > T:A changes, either missense mutations at critical arginine residues or nonsense changes in the 5' half of the gene. The reasons for this odd mutation spectrum were unclear but C:G > T:A changes were not found more often than expected at APC, K-ras, or p53 in the same tumors and we found no specific defects in DNA repair to account for the observations. No colorectal tumor was found to carry two CDC4 mutations predicted to abolish protein function; partial loss of CDC4 function may therefore cause tumorigenesis. The in vitro studies, therefore, did not assess the functional effects of mutant alleles which are found in vivo. CDC4 mutations may be selected primarily to drive progression through the cell cycle although CIN might be an important secondary effect in some cancers. PMID- 16357144 TI - Leukemogenic MLL fusion proteins bind across a broad region of the Hox a9 locus, promoting transcription and multiple histone modifications. AB - Chromosome translocations involving the mixed lineage leukemia gene MLL are associated with aggressive acute leukemias in both children and adults. Leukemogenic MLL fusion proteins delete the MLL SET domain Lys(4) methyltransferase activity and fuse MLL to 1 of >40 different translocation partners. Some MLL fusion proteins involve nuclear proteins that are transcriptional activators, whereas others have transcriptional activating activity but instead dimerize the truncated MLL molecule. Both types of MLL fusion proteins enforce persistent expression of Hox a9 and Meis1, which is pivotal for leukemogenesis through mechanisms that remain obscure. Here, we show that nuclear and dimerizable forms of MLL bind with a similar pattern to the Hox a9 locus that overlaps the distribution of wild-type MLL and deregulate transcription of three isoforms of Hox a9. Induction of MLL fusion protein activity is associated with increased levels of histone acetylation and Lys(4) methylation at Hox target genes. In addition, the MLL-ENL-ER protein, but not dimerized MLL, also induces dimethylation of histone H3 at Lys(79), suggesting alternative mechanisms for transcriptional activation. PMID- 16357145 TI - Inducible IkappaB kinase/IkappaB kinase epsilon expression is induced by CK2 and promotes aberrant nuclear factor-kappaB activation in breast cancer cells. AB - Aberrant activation of nuclear factor-kappaB (NF-kappaB) transcription factors has been implicated in the pathogenesis of breast cancer. We previously showed elevated activity of IkappaB kinase alpha (IKKalpha), IKKbeta, and protein kinase CK2 in primary human breast cancer specimens and cultured cells. A novel inducible IKK protein termed IKK-i/IKKepsilon has been characterized as a potential NF-kappaB activator. Here, we provide evidence that implicates IKK i/IKKepsilon in the pathogenesis of breast cancer. We show IKK-i/IKKepsilon expression in primary human breast cancer specimens and carcinogen-induced mouse mammary tumors. Multiple breast cancer cell lines showed higher levels of IKK i/IKKepsilon and kinase activity compared with untransformed MCF-10F breast epithelial cells. Interestingly, IKK-i/IKKepsilon expression correlated with CK2alpha expression in mammary glands and breast tumors derived from MMTV CK2alpha transgenic mice. Ectopic CK2 expression in untransformed cells led to increased IKK-i/IKKepsilon mRNA and protein levels. Inhibition of CK2alpha via the pharmacologic inhibitor apigenin or upon transfection of a CK2 kinase inactive subunit reduced IKK-i/IKKepsilon levels. Expression of a kinase-inactive IKK-i/IKKepsilon mutant in breast cancer cells reduced NF-kappaB activity as judged by transfection assays of reporters driven either by NF-kappaB elements or the promoters of two NF-kappaB target genes, cyclin D1 and relB. Importantly, the kinase-inactive IKK-i/IKKepsilon mutant reduced the endogenous levels of these genes as well as the ability of breast cancer cells to grow in soft agar or form invasive colonies in Matrigel. Thus, CK2 induces functional IKK-i/IKKepsilon, which is an important mediator of the activation of NF-kappaB that plays a critical role in the pathogenesis of breast cancer. PMID- 16357146 TI - BRCA1 regulates RAD51 function in response to DNA damage and suppresses spontaneous sister chromatid replication slippage: implications for sister chromatid cohesion, genome stability, and carcinogenesis. AB - The breast/ovarian cancer susceptibility proteins BRCA1 and BRCA2 maintain genome stability, at least in part, through a functional role in DNA damage repair. They both colocalize with RAD51 at sites of DNA damage/replication and activate RAD51 mediated homologous recombination repair of DNA double-strand breaks (DSB). Whereas BRCA2 interacts directly with and regulates RAD51, the role of BRCA1 in this process is unclear. However, BRCA1 may regulate RAD51 in response to DNA damage or through its ability to interact with and regulate MRE11/RAD50/NBS1 (MRN) during the processing of DSBs into single-strand DNA (ssDNA) ends, prerequisite substrates for RAD51, or both. To test these hypotheses, we measured the effect of BRCA1 on the competition between RAD51-mediated homologous recombination (gene conversion and crossover) versus RAD51-independent homologous recombination [single-strand annealing (SSA)] for ssDNA at a site-specific chromosomal DSB within a DNA repeat, a substrate for both homologous recombination pathways. Expression of wild-type BRCA1 in BRCA1-deficient human recombination reporter cell lines promoted both gene conversion and SSA but greatly enhanced gene conversion. In addition, BRCA1 also suppressed both spontaneous gene conversion and deletion events, which can arise from either crossover or sister chromatid replication slippage (SCRS), a RAD51-independent process. BRCA1 does not seem to block crossover. From these results, we conclude that (a) BRCA1 regulates RAD51 function in response to the type of DNA damage and (b) BRCA1 suppresses SCRS, suggesting a role for this protein in sister chromatid cohesion/alignment. Loss of such control in response to estrogen-induced DNA damage after BRCA1 inactivation may be a key initial event that triggers genome instability and carcinogenesis. PMID- 16357147 TI - Widespread CXCR4 activation in astrocytomas revealed by phospho-CXCR4-specific antibodies. AB - The chemokine receptor CXCR4 is expressed in many cancers where it may regulate tumor cell growth and migration. The role of CXCR4 in cancer will depend on it being in an activated, signaling state. To better define the significance of CXCR4 expression in cancer, we developed an antibody that can distinguish CXCR4 phosphorylated on serine 339, a residue previously identified as a site for ligand-induced phosphorylation. With this antibody, we investigated the mechanisms of CXCR4 phosphorylation and evaluated the phosphorylation status of CXCR4 in human astrocytomas. In vitro, phosphorylation of serine 339 occurred in response to CXCL12 or epidermal growth factor (EGF) treatment and was increased by protein kinase C activation. In all grades of astrocytomas, CXCR4 was expressed in tumor cells and some endothelial cells, whereas CXCL12 was present in endothelial cells and infiltrating microglia. We found that CXCR4 phosphorylated on serine 339 was present in tumor cells and vascular endothelial cells in all grades of astrocytoma. These data indicate that CXCR4 is expressed and activated in astrocytomas and that phosphorylation of CXCR4 can occur through ligand activation or transactivation via the EGF receptor. These studies extend the potential roles of CXCR4 in cancer to include functions associated with benign (grade 1) tumors. The ability to distinguish phosphorylated CXCR4 will be invaluable for the continued analysis of the role of CXCR4 in cancer and the development of CXCR4 antagonist therapy for patients suffering with primary tumors of the brain and other sites. PMID- 16357148 TI - Methyl CpG-binding domain protein 3 mediates cancer-selective cytotoxicity by histone deacetylase inhibitors via differential transcriptional reprogramming in lung cancer cells. AB - Histone deacetylase inhibitors (HDI) have been reported to inhibit the growth and survival of cancer cells while leaving normal cells untouched. However, the mechanisms underlying this selective cell death are poorly understood. Gene expression analysis revealed that HDI treatment induced up-regulation of p21(WAF1/Cip1) and down-regulation of ErbB2 in cancer cells but not normal cells. Overexpression of p21(WAF1/Cip1) and/or silencing of ErbB2 enhanced cancer cell growth inhibition, suggesting that HDI-induced up-regulation/down-regulation of these genes play critical roles in HDI-induced growth inhibition of cancer cells. Most importantly, we found that the gene silencing factor methyl CpG-binding domain protein 3 (MBD3) was not only released from cancer-selective promoter of the HDI up-regulated p21(WAF1/Cip1) gene but also recruited to that of the HDI down-regulated ErbB2 gene. Furthermore, silencing of MBD3 by small interfering RNA abrogated the HDI-induced gene regulation and growth inhibition in lung cancer but not in normal cells. Together, our results support the critical potential of MBD3 in HDI-induced cancer-selective cell death via cancer differential gene expression. PMID- 16357149 TI - Stepwise neoplastic transformation of a telomerase immortalized fibroblast cell line. AB - We have described recently a human fibroblast cell line immortalized through ectopic telomerase expression (cen3tel), in which the extension of the life span was associated with the appearance of chromosomal aberrations and with the ability to grow in the absence of solid support. As reported in this article, on further propagation in culture, cen3tel cells became neoplastically transformed, being able to form tumors in nude mice. The analysis of the cells, during the gradual transition toward the tumorigenic phenotype, allowed us to trace cellular and molecular changes associated with different phases of transformation. At the stage in which they were able to grow in agar, cen3tel cells had lost contact growth inhibition but still retained the requirement of serum to proliferate and were not tumorigenic in immunocompromised mice. Moreover, they showed a down regulation of the INK4A locus and were resistant to oncogenic Ras-induced senescence but still retained a functional p53. Subsequently, cen3tel cells became tumorigenic, lost p53 function because of a mutation in the DNA-binding motif, and overexpressed c-myc. Interestingly, tumorigenic cells did not carry activating mutations either in the ras proto-oncogenes (H-ras, N-ras, and K-ras) or in B-raf. Cen3tel cells gradually became hyperdiploid but did not display centrosome abnormalities. To our knowledge, cen3tel is the first telomerase immortalized fibroblast line, which became neoplastically transformed. In this system, we could associate a down-regulation of the INK4A locus with anchorage independent growth and with resistance to Ras-induced senescence and link p53 mutations and c-myc overexpression with tumorigenicity. PMID- 16357150 TI - ABCC drug efflux pumps and organic anion uptake transporters in human gliomas and the blood-tumor barrier. AB - Delivery of therapeutic agents to the brain and its neoplasms depends on the presence of membrane transport proteins in the blood-brain barrier and in the target cells. The cellular and subcellular localization of these membrane transporters determines the drug accessibility to the brain and its tumors. We therefore analyzed the expression and localization of six members of the multidrug resistance protein family of ATP-dependent efflux pumps (ABCC1-ABCC6, formerly MRP1-MRP6) and of six organic anion uptake transporters (OATP1A2, OATP1B1, OATP1B3, OATP1C1, OATP2B1, and OATP4A1) in 61 human glioma specimens of different histologic subtypes. Real-time PCRs indicated expressions of ABCC1, ABCC3, ABCC4, and ABCC5. In addition, we detected expressions of the OATP uptake transporter genes SLCO1A2, SLCO1C1, SLCO2B1, and SLCO4A1. At the protein level, however, only OATP1A2 and OATP2B1 were detectable by immunofluorescence microscopy in the luminal membrane of endothelial cells forming the blood-brain barrier and the blood-tumor barrier, but not in the glioma cells. ABCC4 and ABCC5 proteins were the major ABCC subfamily members in gliomas, localized both at the luminal side of the endothelial cells and in the glioma cells of astrocytic tumors and in the astrocytic portions of oligoastrocytomas. These results indicate that expression of ABCC4 and ABCC5 is associated with an astrocytic phenotype, in accordance with their expression in astrocytes and with the higher chemoresistance of astrocytic tumors as compared with oligodendrogliomas. Our data provide a basis for the assessment of the role of uptake transporters and efflux pumps in the accessibility of human gliomas for chemotherapeutic agents. PMID- 16357151 TI - Lysyl oxidase regulates breast cancer cell migration and adhesion through a hydrogen peroxide-mediated mechanism. AB - We have previously shown that lysyl oxidase (LOX) mRNA is up-regulated in invasive breast cancer cells and that catalytically active LOX facilitates in vitro cell invasion. Here we validate our in vitro studies by showing that LOX expression is up-regulated in distant metastatic breast cancer tissues compared with primary cancer tissues. To elucidate the mechanism by which LOX facilitates cell invasion, we show that catalytically active LOX regulates in vitro motility/migration and cell-matrix adhesion formation. Treatment of the invasive breast cancer cell lines, Hs578T and MDA-MB-231, with beta-aminopropionitrile (betaAPN), an irreversible inhibitor of LOX catalytic activity, leads to a significant decrease in cell motility/migration and adhesion formation. Conversely, poorly invasive MCF-7 cells expressing LOX (MCF-7/LOX32-His) showed an increase in migration and adhesion that was reversible with the addition of betaAPN. Moreover, a decrease in activated focal adhesion kinase (FAK) and Src kinase, key proteins involved in adhesion complex turnover, was observed when invasive breast cancer cells were treated with betaAPN. Additionally, FAK and Src activation was increased in MCF-7/LOX32-His cells, which was reversible on betaAPN treatment. Hydrogen peroxide was produced as a by-product of LOX activity and the removal of hydrogen peroxide by catalase treatment in invasive breast cancer cells led to a dose-dependent loss in Src activation. These results suggest that LOX facilitates migration and cell-matrix adhesion formation in invasive breast cancer cells through a hydrogen peroxide-mediated mechanism involving the FAK/Src signaling pathway. These data show the need to target LOX for treatment of aggressive breast cancer. PMID- 16357152 TI - gamma-Aminobutyric acid inhibits cholangiocarcinoma growth by cyclic AMP dependent regulation of the protein kinase A/extracellular signal-regulated kinase 1/2 pathway. AB - We studied the effect of the inhibitory neurotransmitter, gamma-aminobutyric acid (GABA), in the regulation of cholangiocarcinoma growth. We determined the in vitro effect of GABA on the proliferation of the cholangiocarcinoma cell lines (Mz-ChA-1, HuH-28, and TFK-1) and evaluated the intracellular pathways involved. The effect of GABA on migration of Mz-ChA-1 cells was also evaluated. In vivo, Mz ChA-1 cells were s.c. injected in athymic mice, and the effects of GABA on tumor size, tumor cell proliferation, apoptosis, collagen quantity, and the expression of vascular endothelial growth factor-A (VEGF-A) and VEGF-C (cancer growth regulators) were measured after 82 days. GABA decreased in vitro cholangiocarcinoma growth in a time-dependent and dose-dependent manner, by both cyclic AMP/protein kinase A- and D-myo-inositol-1,4,5-thriphosphate/Ca(2+) dependent pathways, leading to down-regulation of extracellular signal-regulated kinase 1/2 phosphorylation. Blocking of GABA(A), GABA(B), and GABA(C) receptors prevented GABA inhibition of cholangiocarcinoma proliferation. GABA inhibited Mz ChA-1 cell migration and, in vivo, significantly decreased tumor volume, tumor cell proliferation, and VEGF-A/C expression whereas increasing apoptosis compared with controls. An increase in collagen was evident in GABA-treated tumors. GABA decreases biliary cancer proliferation and reduces the metastatic potential of cholangiocarcinoma. GABA may represent a therapeutic agent for patients affected by malignancies of the biliary tract. PMID- 16357153 TI - Cyclooxygenase-2 inhibition sensitizes human colon carcinoma cells to TRAIL induced apoptosis through clustering of DR5 and concentrating death-inducing signaling complex components into ceramide-enriched caveolae. AB - Cyclooxygenase-2 (COX-2) is up-regulated in human colon carcinomas, and its inhibition is associated with a reduction in tumorigenesis and a promotion of apoptosis. However, the mechanisms responsible for the antitumor effects of COX-2 inhibitors and how COX-2 modulates apoptotic signaling have not been clearly defined. We have shown that COX-2 inhibition sensitizes human colon carcinoma cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by inducing clustering of the TRAIL receptor DR5 at the cell surface and the redistribution of the death-inducing signaling complex components (DR5, FADD, and procaspase-8) into cholesterol-rich and ceramide-rich domains known as caveolae. This process requires the accumulation of arachidonic acid and sequential activation of acid sphingomyelinase for the generation of ceramide within the plasma membrane outer leaflet. The current study highlights a novel mechanism to circumvent colorectal carcinoma cell resistance to TRAIL-mediated apoptosis using COX-2 inhibitors to manipulate the lipid metabolism within the plasma membrane. PMID- 16357154 TI - Development of Ewing's sarcoma from primary bone marrow-derived mesenchymal progenitor cells. AB - Ewing's sarcoma is a member of Ewing's family tumors (EFTs) and the second most common solid bone and soft tissue malignancy of children and young adults. It is associated in 85% of cases with the t(11;22)(q24:q12) chromosomal translocation that generates fusion of the 5' segment of the EWS gene with the 3' segment of the ETS family gene FLI-1. The EWS-FLI-1 fusion protein behaves as an aberrant transcriptional activator and is believed to contribute to EFT development. However, EWS-FLI-1 induces growth arrest and apoptosis in normal fibroblasts, and primary cells that are permissive for its putative oncogenic properties have not been discovered, hampering basic understanding of EFT biology. Here, we show that EWS-FLI-1 alone can transform primary bone marrow-derived mesenchymal progenitor cells and generate tumors that display hallmarks of Ewing's sarcoma, including a small round cell phenotype, expression of EFT-associated markers, insulin like growth factor-I dependence, and induction or repression of numerous EWS-FLI-1 target genes. These observations provide the first identification of candidate primary cells from which EFTs originate and suggest that EWS-FLI-1 expression may constitute the initiating event in EFT pathogenesis. PMID- 16357155 TI - Inhibition of DNA methylation sensitizes glioblastoma for tumor necrosis factor related apoptosis-inducing ligand-mediated destruction. AB - Life expectancy of patients affected by glioblastoma multiforme is extremely low. The therapeutic use of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been proposed to treat this disease based on its ability to kill glioma cell lines in vitro and in vivo. Here, we show that, differently from glioma cell lines, glioblastoma multiforme tumors were resistant to TRAIL stimulation because they expressed low levels of caspase-8 and high levels of the death receptor inhibitor PED/PEA-15. Inhibition of methyltransferases by decitabine resulted in considerable up-regulation of TRAIL receptor-1 and caspase 8, down-regulation of PED/PEA-15, inhibition of cell growth, and sensitization of primary glioblastoma cells to TRAIL-induced apoptosis. Exogenous caspase-8 expression was the main event able to restore TRAIL sensitivity in primary glioblastoma cells. The antitumor activity of decitabine and TRAIL was confirmed in vivo in a mouse model of glioblastoma multiforme. Evaluation of tumor size, apoptosis, and caspase activation in nude mouse glioblastoma multiforme xenografts showed dramatic synergy of decitabine and TRAIL in the treatment of glioblastoma, whereas the single agents were scarcely effective in terms of reduction of tumor mass, apoptosis induction, and caspase activation. Thus, the combination of TRAIL and demethylating agents may provide a key tool to overcome glioblastoma resistance to therapeutic treatments. PMID- 16357156 TI - High expression of ErbB family members and their ligands in lung adenocarcinomas that are sensitive to inhibition of epidermal growth factor receptor. AB - Recent findings in tumor biopsies from lung adenocarcinoma patients suggest that somatic mutations in the genes encoding epidermal growth factor receptor (EGFR) and Kirsten ras (KRAS) confer sensitivity and resistance, respectively, to EGFR inhibition. Here, we provide evidence that these genetic mutations are not sufficient to modulate the biological response of lung adenocarcinoma cells to EGFR inhibition. We found high expression of ErbB family members, ErbB ligands, or both in three models that were sensitive to EGFR inhibition, including alveolar epithelial neoplastic lesions in mice that develop lung adenocarcinoma by oncogenic KRAS, human lung adenocarcinoma cell lines, and tumor biopsies from lung adenocarcinoma patients. Thus, lung adenocarcinoma cells that depend on EGFR for survival constitutively activate the receptor through a combination of genetic mutations and overexpression of EGFR dimeric partners and their ligands. PMID- 16357157 TI - The critical role of 15-lipoxygenase-1 in colorectal epithelial cell terminal differentiation and tumorigenesis. AB - Terminal differentiation is an important event for maintaining normal homeostasis in the colorectal epithelium, and the loss of apoptosis is an important mechanism underlying colorectal tumorigenesis. The very limited current data on the role of lipoxygenase (LOX) metabolism in tumorigenesis suggests that the oxidative metabolism of linoleic and arachidonic acid possibly shifts from producing antitumorigenic 15-LOX-1 and 15-LOX-2 products to producing protumorigenic 5-LOX and 12-LOX products. We examined whether this shift occurs in vitro in the human colon cancer cell line Caco-2 in association with the loss of terminal differentiation and apoptosis, or in vivo during the formation of colorectal adenomas in patients with familial adenomatous polyposis (FAP). Restoring terminal differentiation and apoptosis of Caco-2 cells increased the mRNA levels of 5-LOX, 15-LOX-2, and 15-LOX-1, but the only significant increases in protein expression and enzymatic activity were of 15-LOX-1. In FAP patients, 15-LOX-1 expression and activity were significantly down-regulated in adenomas (compared with paired nonneoplastic epithelial mucosa), whereas 5-LOX and 15-LOX-2 protein expressions and enzymatic activities were not. We conducted a validation study with immunohistochemical testing in a second group of FAP patients; 15-LOX-1 expression was down-regulated in colorectal adenomas (compared with nonneoplastic epithelial mucosa) in 87% (13 of 15) of this group. We confirmed the mechanistic relevance of these findings by demonstrating that ectopically restoring 15-LOX-1 expression reestablished apoptosis in Caco-2 cells. Therefore, 15-LOX-1 down regulation rather than a shift in the balance of LOXs is likely the dominant alteration in LOX metabolism which contributes to colorectal tumorigenesis by repressing apoptosis. PMID- 16357158 TI - Expression of endogenous oncogenic V600EB-raf induces proliferation and developmental defects in mice and transformation of primary fibroblasts. AB - Mutations of the human B-RAF gene are detected in approximately 8% of cancer samples, primarily in cutaneous melanomas (70%). The most common mutation (90%) is a valine-to-glutamic acid mutation at residue 600 (V600E; formerly V599E according to previous nomenclature). Using a Cre/Lox approach, we have generated a conditional knock-in allele of (V600E)B-raf in mice. We show that widespread expression of (V600E)B-Raf cannot be tolerated in embryonic development, with embryos dying approximately 7.5 dpc. Directed expression of mutant (V600E)B-Raf to somatic tissues using the IFN-inducible Mx1-Cre mouse strain induces a proliferative disorder and bone marrow failure with evidence of nonlymphoid neoplasia of the histiocytic type leading to death within 4 weeks of age. However, expression of mutant B-Raf does not alter the proliferation profile of all somatic tissues. In primary mouse embryonic fibroblasts, expression of endogenous (V600E)B-Raf induces morphologic transformation, increased cell proliferation, and loss of contact inhibition. Thus, (V600E)B-Raf is able to induce several hallmarks of transformation in some primary mouse cells without evidence for the involvement of a cooperating oncogene or tumor suppressor gene. PMID- 16357159 TI - Headpin: a serpin with endogenous and exogenous suppression of angiogenesis. AB - Headpin is a novel serine proteinase inhibitor (serpin) with constitutive mRNA expression in histologically normal oral mucosa but with lost or down-regulated expression in head and neck squamous cell carcinoma. Several serpin family members are similarly lost in multiple cancer types and hold tumor suppressor functions including the inhibition of angiogenesis. However, the functional significance for the loss of headpin expression in cancer is not known. Using immunohistochemical analysis of invasive squamous cell carcinoma and matched normal squamous mucosa of patient specimens, headpin expression was lost or down regulated in the vast majority of tumor specimens. We investigated the functions of exogenous recombinant headpin and endogenously expressed headpin related to angiogenesis. In a rat corneal assay of neovascularization, recombinant headpin protein blocked in vivo angiogenesis mediated by interleukin 8 (IL-8) and vascular endothelial growth factor (VEGF). In assays of cellular events in angiogenesis, headpin blocked the invasion, migration, and tube formation of endothelial cells. In light of our findings of nuclear subcellular localization of headpin, we investigated the expression and secretion of angiogenic factors and found reduced mRNA, protein, and promoter activities of IL-8 and VEGF. Finally, using a murine flank tumor model, headpin expression reduced growth and microvessel density in tumors derived from headpin-expressing UMSCC1 cells relative to those from vector control cells. These findings of nuclear regulatory functions of a serpin in the inhibition of angiogenesis bring new understanding to the cellular and molecular mechanisms of serpins. Therefore, this novel serpin targets diverse mechanisms against tumor angiogenesis on which to base therapeutic strategies. PMID- 16357160 TI - Bortezomib inhibits PKR-like endoplasmic reticulum (ER) kinase and induces apoptosis via ER stress in human pancreatic cancer cells. AB - Bortezomib (Velcade, formerly known as PS-341) is a boronic acid dipeptide derivative that is a selective and potent inhibitor of the proteasome. We hypothesized that proteasome inhibition would lead to an accumulation of misfolded proteins in the cell resulting in endoplasmic reticulum (ER) stress. The ability of bortezomib to induce ER stress and the unfolded protein response was investigated in a human pancreatic cancer cell line, L3.6pl. Bortezomib increased expression of ER stress markers, CHOP and BiP, but inhibited PKR-like ER kinase and subsequent phosphorylation of eukaryotic initiation factor 2alpha (eif2alpha), both of which are key events in translational suppression. These effects resulted in an accumulation of ubiquitylated proteins leading to protein aggregation and proteotoxicity. Peptide inhibitor or small interfering RNA targeting ER-resident caspase-4 blocked DNA fragmentation, establishing a central role for caspase-4 in bortezomib-induced cell death. The translation inhibitor cycloheximide abrogated bortezomib-induced protein aggregation, caspase-4 processing, and all other characteristics of apoptosis. Because malignant cells have higher protein synthesis rates than normal cells, they may be more prone to protein aggregation and proteotoxicity and possess increased sensitivity to bortezomib-induced apoptosis. Taken together, the results show that bortezomib induces a unique type of ER stress compared with other ER stress agents characterized by an absence of eif2alpha phosphorylation, ubiquitylated protein accumulation, and proteotoxicity. PMID- 16357161 TI - Tumor cell plasticity in Ewing sarcoma, an alternative circulatory system stimulated by hypoxia. AB - A striking feature of Ewing sarcoma is the presence of blood lakes lined by tumor cells. The significance of these structures, if any, is unknown. Here, we report that the extent of blood lakes correlates with poor clinical outcomes, whereas variables of angiogenesis do not. We also show that Ewing sarcoma cells form vessel-like tubes in vitro and express genes associated with vasculogenic mimicry. In tumor models, we show that there is blood flow through the blood lakes, suggesting that these structures in Ewing sarcoma contribute to the circulation. Furthermore, we present evidence that reduced oxygen tension may be instrumental in tube formation by plastic tumor cells. The abundant presence of these vasculogenic structures, in contrast to other tumor types, makes Ewing sarcoma the ideal model system to study these phenomena. The results suggest that optimal tumor treatment may require targeting of these structures in combination with prevention of angiogenesis. PMID- 16357162 TI - Apoptotic cells initiate endothelial cell sprouting via electrostatic signaling. AB - Angiogenesis, the development of new blood vessels from preexisting vessels, is crucial to tissue growth, repair, and maintenance. This process begins with the formation of endothelial cell sprouts followed by the proliferation and migration of neighboring endothelial cells along the preformed extensions. The initiating event and mechanism of sprouting is not known. We show that the phenotypic expression of negatively charged membrane surface in apoptotic cells initiates the formation of directional endothelial cell sprouts that extend toward the dying cells by a mechanism that involves endothelial cell membrane hyperpolarization and cytoskeleton reorganization but is independent of diffusible molecules. PMID- 16357163 TI - The G691S RET polymorphism increases glial cell line-derived neurotrophic factor induced pancreatic cancer cell invasion by amplifying mitogen-activated protein kinase signaling. AB - Mutations of the RET proto-oncogene are responsible for several inherited human diseases and may function as genetic modifiers of the disease. However, the role of RET mutations in pancreatic cancer has not been studied. Expression of the glial cell line-derived neurotrophic factor (GDNF) receptors RET and GDNF family receptor alpha1 (GFRalpha1) in human pancreatic cancer cells was determined by Western blot, immunofluorescence, and flow cytometry. The effect of GDNF on cell proliferation and invasion was assessed. Small interfering RNA and antibodies were used to evaluate the involvement of RET. The G691S RET polymorphism was analyzed by sequencing and restriction analysis. The modifying effect of G691S RET on GDNF-induced invasion and mitogen-activated protein kinase (MAPK) signaling was evaluated. Transfection studies with wild-type and mutated RET determined the functional role of the G691S polymorphism. Pancreatic cancer specimens and matched tissues were analyzed for the presence of the G691S RET polymorphism. GDNF receptors were found on all cell lines. GDNF increased pancreatic cancer cell proliferation and invasion, which was mediated by RET. The effect of GDNF was more profound in cells with the G691S RET polymorphism (P < 0.01). G691S RET correlated with an enhanced activation of the downstream extracellular signal-regulated kinase pathway. Overexpression of G691S RET increased pancreatic cancer cell invasion. The G691S RET polymorphism was also detected in human pancreatic tumors and represented a somatic mutation in some patients. These findings indicate that the G691S RET single nucleotide polymorphism may directly correlate with the aggressive growth of pancreatic cancers and may function as a genetic modifier or even low-penetrance gene. PMID- 16357165 TI - GoldIII porphyrin 1a induced apoptosis by mitochondrial death pathways related to reactive oxygen species. AB - Apoptosis is a tightly controlled multistep mechanism of cell death, and mitochondria are considered to play a central role in this process. Mitochondria initiate two distinct apoptosis pathways, one caspase-dependent and the other caspase-independent. In addition, mitochondrial production of reactive oxygen species (ROS) seems to play a role in cell death. Most chemotherapeutic agents induce apoptosis through at least one of these pathways. The post-initiation mechanisms of gold(III) porphyrin 1a were investigated in this study. HONE1 cells exposed to gold(III) porphyrin 1a underwent apoptosis after 24 hours. Functional proteomic studies revealed the alteration of several cytoplasmic protein expressions in HONE1 cells after treatment with the drug. These proteins include enzymes participating in energy production and proteins involved in cellular redox balance. There was a quick attenuation of mitochondrial membrane potential (DeltaPsi(m)) with the alterations of Bcl-2 family proteins, the release of cytochrome c, and apoptosis-inducing factor (AIF) following gold(III) porphyrin 1a treatment. Cytochrome c in turn activated caspase-9 and caspase-3. Cotreatment with caspase inhibitor (zVAD-fmk) showed that the activated caspases worked in conjunction with AIF-initiated apoptosis pathways. Further study showed that ROS played a part in gold(III) porphyrin 1a-induced apoptosis by regulating DeltaPsi(m). In summary, gold(III) porphyrin 1a induced apoptosis through both caspase-dependent and caspase-independent mitochondrial pathways, and intracellular oxidation affected gold(III) porphyrin 1a-induced apoptosis. These results support a role for gold(III) porphyrin 1a as a promising anticancer drug lead and as a possible novel therapeutic agent directed toward the mitochondria. PMID- 16357164 TI - Dentin matrix protein 1 enhances invasion potential of colon cancer cells by bridging matrix metalloproteinase-9 to integrins and CD44. AB - The up-regulation of various matrix metalloproteinases (MMP), certain cell receptors such as integrins and CD44, and the SIBLING family of integrin-binding glycophosphoproteins have been reported separately and in various combinations for many types of tumors. The mechanisms by which these different proteins may be interacting and enhancing the ability of a cancer cell to survive and metastasize have become an interesting issue in cancer biology. Dentin matrix protein 1 (DMP1) has been known for a number of years to bind to CD44 and ArgGlyAsp sequence-dependent integrins. This SIBLING was recently shown to be able to specifically bind and activate proMMP-9 and to make MMP-9 much less sensitive to inhibition by tissue inhibitors of metalloproteinases and synthetic inhibitors. In this study, we used a modified Boyden chamber assay to show that DMP1 enhanced the invasiveness of the MMP-9 expressing colon cancer cell line, SW480, through Matrigel in a dose-dependant manner. DMP1 (100 nmol/L) increased invasion 4-fold over controls (86.1 +/- 13.9 versus 22.3 +/- 9.8, P < 0.001). The enhanced invasive potential required the presence of MMP-9 and at least one of the cell surface receptors, CD44, alpha(v)beta(3), or alpha(v)beta(5) integrin. The bridging of MMP-9 to the cell surface receptors was shown by both pull-down and fluorescence activated cell sorting experiments. Because all of these proteins were also shown by immunohistochemistry to be expressed in serial sections of a colon adenocarcinoma, we have hypothesized that the MMP-9/DMP1/cell surface complexes observed to enhance cell invasion in vitro may be aiding metastatic events in vivo. PMID- 16357166 TI - Context-dependent hormone-refractory progression revealed through characterization of a novel murine prostate cancer cell line. AB - Insights into the molecular basis of hormone-refractory prostate cancer have principally relied on human prostate cancer cell lines, all of which were derived from patients who had already failed hormonal therapy. Recent progress in developing genetically engineered mouse prostate cancer models provides an opportunity to isolate novel cell lines from animals never exposed to hormone ablation, avoiding any potential bias conferred by the selective pressure of the castrate environment. Here we report the isolation of such a cell line (Myc-CaP) from a c-myc transgenic mouse with prostate cancer. Myc-CaP cells have an amplified androgen receptor gene despite no prior exposure to androgen withdrawal and they retain androgen-dependent transgene expression as well as androgen dependent growth in soft agar and in mice. Reexpression of c-Myc from a hormone independent promoter rescues growth in androgen-depleted agar but not in castrated mice, showing a clear distinction between the molecular requirements for hormone-refractory growth in vitro versus in vivo. Myc-CaP cells represent a unique reagent for dissecting discreet steps in hormone-refractory prostate cancer progression and show the general utility of using genetically engineered mouse models for establishing new prostate cancer cell lines. PMID- 16357167 TI - Novel role for PDEF in epithelial cell migration and invasion. AB - Cell migration and invasion are two critical cellular processes that are often deregulated during tumorigenesis. To identify factors that contribute to oncogenic progression by stimulating cell migration, we conducted a powerful retroviral based migration screen using an MCF7 cDNA library and the immortalized human breast epithelial cell line MCF-10A. We identified prostate derived Ets factor (PDEF), an Ets transcription factor that is overexpressed in both prostate and breast carcinoma, as a candidate promigratory gene from this screen. Whereas PDEF induced limited motility of MCF-10A cells, coexpression of PDEF with the receptor tyrosine kinases (RTK) ErbB2 and colony-stimulating factor receptor (CSF 1R)/CSF-1 significantly enhanced MCF-10A motility. Furthermore, cells coexpressing PDEF with either ErbB2 or CSF-1R/CSF-1 induced a dramatic invasive phenotype in three-dimensional cultures. Constitutive activation of the extracellular signal-regulated kinase (ERK) pathway also enhanced PDEF-induced motility and invasion, suggesting that activation of the ERK/mitogen-activated protein kinase by ErbB2 and CSF-1R/CSF-1 can cooperate with PDEF to promote motility and invasion. Furthermore, PDEF promoted anchorage-independent growth of ErbB2 and CSF-1R/CSF-1-expressing cells. Using laser capture microdissection, we also found that PDEF mRNA is overexpressed in breast tumor epithelia throughout tumor progression. Taken together, these findings suggest that the transcription factor PDEF may play an important role in breast tumorigenesis and that PDEF overexpression may be particularly significant in tumors that exhibit activation of oncogenic RTKs such as ErbB2 and CSF-1R. PMID- 16357168 TI - Prognostic and functional significance of thromboxane synthase gene overexpression in invasive bladder cancer. AB - Thromboxane synthase (TXAS) is one of the enzymes downstream from cyclooxygenase 2 and catalyzes the synthesis of thromboxane A(2) (TXA(2)). TXAS was among the genes we identified based on its overexpression in invasive bladder tumors. TXAS is overexpressed in common forms of bladder tumors: 69 of 97 (71.1%) transitional cell carcinoma (TCC), 38 of 53 (71.6%) squamous cell carcinoma, and 5 of 11 (45.5%) adenocarcinoma relative to nontumor tissue. Overall, 112 of 161 (69.5%) invasive tumors exhibited elevated expression. Significantly, patients with tumors having >4-fold levels of TXAS expression showed significant statistical evidence of lower overall survival expressed by the estimated hazard ratio of 2.74 with P = 0.009 in Cox's regression analysis. TXAS mRNA expression was found to be an independent prognostic marker for patients with bladder cancer. Treatment of bladder cancer cell lines (T24 and TCC-SUP) with TXAS inhibitors and TXA(2) (TP) receptor antagonists reduced cell growth, migration, and invasion, whereas TP agonists stimulated cell migration and invasion. The positive correlation between elevated TXAS expression and shorter patient survival supports a potential role for TXAS-regulated pathways in tumor invasion and metastases and suggests that modulation of the TXAS pathway may offer a novel therapeutic approach. PMID- 16357169 TI - Differentiation of androgen-independent prostate cancer PC-3 cells is associated with increased nuclear factor-kappaB activity. AB - Recently, we have reported that inosine 5'-monophosphate dehydrogenase inhibitors, such as mycophenolic acid (MPA), induce the differentiation of PC-3 cells, which are derived from a human androgen-independent prostate cancer, into cells with a phenotype resembling maturing prostate secretory cells. Here, we describe such differentiation induced by the histone deacetylase inhibitor tributyrin. The maturation was defined by cytoplasmic vacuole production and induction of CD10, CD46, CD55, GRP78, keratin 17, and zinc-alpha-2-glycoprotein. To identify additional genes associated with tributyrin-induced PC-3 cell differentiation and to gain some insight into the mechanism that underlies this differentiation, we have, by means of microarray analyses, compared tributyrin induced gene expression patterns with those of MPA, which initiates PC-3 cell differentiation by a dissimilar mode of action. We suggested that genes induced by both tributyrin and MPA would be most likely associated with differentiation rather than with the unique action of each particular inducer. Our results indicated that tributyrin or MPA induced the expression of a large number of common genes, including genes known or assumed to be NF-kappaB dependent. The NF kappaB dependency of a group of these genes, which included the PC-3 cell differentiation marker keratin 17, was confirmed by using two common NF-kappaB activation inhibitors, Bay11-082 and TMB-8, and p65 subunit of NF-kappaB complex specific small interfering RNA. Taken together, our results implicate both NF kappaB-dependent and NF-kappaB-independent genes in the processes leading to PC-3 cell differentiation induced by tributyrin and MPA. PMID- 16357170 TI - Hypoxia-induced down-regulation of BRCA1 expression by E2Fs. AB - Decreased BRCA1 expression in the absence of genetic mutation is observed frequently in sporadic cancers of the breast and other sites, although little is known regarding the mechanisms by which the expression of this gene can be repressed. Here, we show that activating and repressive E2Fs simultaneously bind the BRCA1 promoter at two adjacent E2F sites in vivo, and that hypoxia induces a dynamic redistribution of promoter occupancy by these factors resulting in the transcriptional repression of BRCA1 expression. Functionally, we show that hypoxia is associated with impaired homologous recombination, whereas the nonhomologous end-joining (NHEJ) repair pathway is unaffected under these conditions. Repression of BRCA1 expression by hypoxia represents an intriguing mechanism of functional BRCA1 inactivation in the absence of genetic mutation. We propose that hypoxia-induced decreases in BRCA1 expression and consequent suppression of homologous recombination may lead to genetic instability by shifting the balance between the high-fidelity homologous recombination pathway and the error-prone NHEJ pathway of DNA repair. Furthermore, these findings provide a novel link between E2Fs and the transcriptional response to hypoxia and provide insight into the mechanisms by which the tumor microenvironment can contribute to genetic instability in cancer. PMID- 16357171 TI - The shed ectodomain of Nr-CAM stimulates cell proliferation and motility, and confers cell transformation. AB - Nr-CAM, a cell-cell adhesion molecule of the immunoglobulin-like cell adhesion molecule family, known for its function in neuronal outgrowth and guidance, was recently identified as a target gene of beta-catenin signaling in human melanoma and colon carcinoma cells and tissue. Retrovirally mediated transduction of Nr CAM into fibroblasts induces cell motility and tumorigenesis. We investigated the mechanisms by which Nr-CAM can confer properties related to tumor cell behavior and found that Nr-CAM expression in NIH3T3 cells protects cells from apoptosis in the absence of serum by constitutively activating the extracellular signal regulated kinase and AKT signaling pathways. We detected a metalloprotease mediated shedding of Nr-CAM into the culture medium of cells transfected with Nr CAM, and of endogenous Nr-CAM in B16 melanoma cells. Conditioned medium and purified Nr-CAM-Fc fusion protein both enhanced cell motility, proliferation, and extracellular signal-regulated kinase and AKT activation. Moreover, Nr-CAM was found in complex with alpha4beta1 integrins in melanoma cells, indicating that it can mediate, in addition to homophilic cell-cell adhesion, heterophilic adhesion with extracellular matrix receptors. Suppression of Nr-CAM levels by small interfering RNA in B16 melanoma inhibited the adhesive and tumorigenic capacities of these cells. Stable expression of the Nr-CAM ectodomain in NIH3T3 cells conferred cell transformation and tumorigenesis in mice, suggesting that the metalloprotease-mediated shedding of Nr-CAM is a principal route for promoting oncogenesis by Nr-CAM. PMID- 16357172 TI - Papillomavirus E2 protein induces expression of the matrix metalloproteinase-9 via the extracellular signal-regulated kinase/activator protein-1 signaling pathway. AB - Papillomaviruses are involved in the development of cancers of the female cervix, head and neck, and skin. An excellent model to study papillomavirus-induced tumor induction and progression is the New Zealand White rabbit, where the skin is infected with the cottontail rabbit papillomavirus (CRPV). This leads to the formation of benign tumors that progress into invasive and metastasizing carcinomas without the need for cofactors. We have shown previously that specific mutations in the transactivation domain of the transcription/replication factor E2 cause a dramatic loss in the tumor induction efficiency of the viral genome and a major deficiency in tumor progression as we show now. By comparing wild type (WT) and mutant E2-induced skin tumors, we found high levels of matrix metalloproteinase-9 (MMP-9) protein and transcripts in WT CRPV-E2-induced tumors in contrast to certain mutant CRPV-E2-induced papillomas and normal uninfected skin. Stable cell lines and reporter assays revealed that E2 from different papillomavirus types is able to transactivate the MMP-9 promoter via the promoter proximal activator protein-1 (AP-1) site as shown in reporter gene assays with mutant MMP-9 promoter constructs. Furthermore, WT E2 but not mutant E2 strongly transactivated a minimal promoter reporter construct with multiple AP-1 sites. The MMP-9 protein induced in cells expressing E2 degrades collagen matrices as measured in Matrigel-based invasion/mobility assays. E2-induced MMP-9 expression can be blocked by a chemical inhibitor of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase 1 (PD 098059), suggesting that E2 activates the MAPK/ERK signaling pathway, which is further supported by the induction of ERK1 in CRPV-E2-transfected cells. PMID- 16357173 TI - Hypoxia-regulated expression of attenuated diphtheria toxin A fused with hypoxia inducible factor-1alpha oxygen-dependent degradation domain preferentially induces apoptosis of hypoxic cells in solid tumor. AB - Tumor cells in hypoxic areas of solid tumors are resistant to conventional chemotherapy and radiotherapy and thus are obstacles of cancer therapy. We report here the feasibility of applying hypoxia-regulated expression of diphtheria toxin A (DT-A) for killing hypoxic tumor cells. The expression vector was constructed to express DT-A fused with hypoxia-inducible factor-1alpha (HIF-1alpha) oxygen dependent degradation (ODD) domain under the control of vascular endothelial growth factor gene promoter and contain erythropoietin mRNA-binding protein (ERBP)-binding sequence downstream of the DT-A/ODD sequence. In vitro ubiquitination assay showed that DT-A/ODD, but not DT-A, was ubiquitinated as efficient as HIF-1alpha under normoxic conditions in a von Hippel-Lindau- and oxygen-dependent manner. DT-A/ODD exhibited a comparable translation inhibitory activity to DT-A. ERBP-binding sequence was effective in stabilizing mRNA under hypoxic conditions in various cell types. Transfection of the vector expressing DT-A/ODD into high-metastatic Lewis lung carcinoma (3LL) A11 cells resulted in induction of apoptosis independently of hypoxia, probably due to its extreme toxicity. However, transfection of the vector expressing attenuated DT A(W153F)/ODD or DT-A(H21A)/ODD resulted in a hypoxia-dependent induction of apoptosis. Liposomal gene transfer of the vector encoding DT-A(W153F)/ODD induced apoptosis in hypoxic, but not in normoxic, areas of solid tumors established by A11 variant cells with higher resistance to hypoxia-induced apoptosis and inhibited the growth of hypoxic tumors established by 3LL-P29 cells. These results suggest that hypoxia-regulated expression of attenuated DT-A(W153F)/ODD fusion protein is potentially of use for killing hypoxic tumor cells with minimizing the damage to normoxic normal tissues. PMID- 16357174 TI - Epidermal growth factor receptor-targeted immunoliposomes significantly enhance the efficacy of multiple anticancer drugs in vivo. AB - We previously reported the development of epidermal growth factor receptor (EGFR) targeted immunoliposomes that bind and internalize in tumor cells which overexpress EGFR and/or mutant EGFR variant III (EGFRvIII), enabling intracellular delivery of potent anticancer agents in vitro. We now describe in vivo proof-of-concept for this approach for the delivery of multiple anticancer drugs in EGFR-overexpressing tumor models. Anti-EGFR immunoliposomes were constructed modularly with Fab' fragments of cetuximab (IMC-C225), covalently linked to liposomes containing probes and/or anticancer drugs. Pharmacokinetic and biodistribution studies confirmed long circulation times (t(1/2) = 21 hours) and efficient accumulation in tumors (up to 15% ID/g) irrespective of the presence of the targeting ligand. Although total accumulations of anti-EGFR immunoliposomes and nontargeted liposomes in EGFR-overexpressing tumors were comparable, only immunoliposomes internalized extensively within tumor cells (92% of analyzed cells versus <5% for nontargeted liposomes), indicating different mechanisms of delivery at the cellular level. In vivo therapy studies in a series of xenograft models featuring overexpression of EGFR and/or EGFRvIII showed the superiority of immunoliposomal delivery of encapsulated drugs, which included doxorubicin, epirubicin, and vinorelbine. For each of these drugs, anti-EGFR immunoliposome delivery showed significant antitumor effects and was significantly superior to all other treatments, including the corresponding free or liposomal drug (P < 0.001-0.003). We conclude that anti-EGFR immunoliposomes provide efficient and targeted drug delivery of anticancer compounds and may represent a useful new treatment approach for tumors that overexpress the EGFR. PMID- 16357175 TI - Cancer cells regulate lymphocyte recruitment and leukocyte-endothelium interactions in the tumor-draining lymph node. AB - The physiologic function of the secondary lymphoid organs to recruit large numbers of naive lymphocytes increases the probability that antigens encounter their rare, sometimes unique, specific T lymphocytes and initiate a specific immune response. In peripheral lymph nodes (LNs), this recruitment is a multistep process, initiated predominantly within the high endothelial venules (HEVs), beginning with rolling and chemokine-dependent firm adhesion of the lymphocytes on the venular endothelium surface. We report here that, in C57BL/6 mice, the recruitment of naive lymphocytes is impaired in LNs draining a B16 melanoma tumor. Intravital microscopy analysis of the tumor-draining LNs revealed that this effect is associated with an important defect in lymphocyte adhesion in the HEVs and a progressive decrease in the expression of the LN chemokine CCL21. In parallel with these effects, the tumor up-regulated, essentially through a P selectin-dependent mechanism, the rolling and sticking of circulating polymorphonuclear cells within the LN low-order venules where few rolling and sticking events are usually observed. These effects of the tumor were independent of the presence of metastasis into the LN and occurred as long as the tumor developed. Together, these results indicate that the tumor proximity disturbs the LN physiology by modifying the molecular, spatial, and cellular rules that usually control leukocyte-endothelium interactions into the peripheral LNs. In addition, they emphasize a new role for the low-order venules of the peripheral LNs, which compared with the HEVs, seem to be the preferential port of entry for cells linked to inflammatory processes. PMID- 16357176 TI - Endothelin-1 promotes epithelial-to-mesenchymal transition in human ovarian cancer cells. AB - Despite considerable efforts to improve early detection and advances in chemotherapy, metastatic relapses remain a major challenge in the management of ovarian cancer. The endothelin A receptor (ET(A)R)/endothelin-1 (ET-1) axis has been shown to have a significant role in ovarian carcinoma by promoting tumorigenesis. Here we show that the ET-1/ET(A)R autocrine pathway drives epithelial-to-mesenchymal transition (EMT) in ovarian tumor cells by inducing a fibroblastoid and invasive phenotype, down-regulation of E-cadherin, increased levels of beta-catenin, Snail, and other mesenchymal markers, and suppression of E-cadherin promoter activity. Activation of ET(A)R by ET-1 triggers an integrin linked kinase (ILK)-mediated signaling pathway leading to glycogen synthase kinase-3beta (GSK-3beta) inhibition, Snail and beta-catenin stabilization, and regulation of transcriptional programs that control EMT. Transfection of dominant negative ILK or exposure to an ILK inhibitor suppresses the ET-1-induced phosphorylation of GSK-3beta as well as Snail and beta-catenin protein stability, activity, and invasiveness, indicating that ET-1/ET(A)R-induced EMT-promoting effects depend on ILK. ET(A)R blockade by specific antagonists or reduction by ET(A)R RNA interference reverses EMT and cell invasion by inhibiting autocrine signaling pathways. In ovarian carcinoma xenografts, ABT-627, a specific ET(A)R antagonist, suppresses EMT determinants and tumor growth. In human ovarian cancers, ET(A)R expression is associated with E-cadherin down-regulation, N cadherin expression, and tumor grade. Collectively, these findings provide evidence of a critical role for the ET-1/ET(A)R axis during distinct steps of ovarian carcinoma progression and identify novel targets of therapeutic intervention. PMID- 16357177 TI - Bortezomib sensitizes pancreatic cancer cells to endoplasmic reticulum stress mediated apoptosis. AB - Bortezomib (PS-341, Velcade) is a potent and selective inhibitor of the proteasome that is currently under investigation for the treatment of solid malignancies. We have shown previously that bortezomib has activity in pancreatic cancer models and that the drug induces endoplasmic reticulum (ER) stress but also suppresses the unfolded protein response (UPR). Because the UPR is an important cytoprotective mechanism, we hypothesized that bortezomib would sensitize pancreatic cancer cells to ER stress-mediated apoptosis. Here, we show that bortezomib promotes apoptosis triggered by classic ER stress inducers (tunicamycin and thapsigargin) via a c-Jun NH(2)-terminal kinase (JNK)-dependent mechanism. We also show that cisplatin stimulates ER stress and interacts with bortezomib to increase ER dilation, intracellular Ca(2+) levels, and cell death. Importantly, combined therapy with bortezomib plus cisplatin induced JNK activation and apoptosis in orthotopic pancreatic tumors resulting in a reduction in tumor burden. Taken together, our data establish that bortezomib sensitizes pancreatic cancer cells to ER stress-induced apoptosis and show that bortezomib strongly enhances the anticancer activity of cisplatin. PMID- 16357178 TI - Sphingosine kinase-1 as a chemotherapy sensor in prostate adenocarcinoma cell and mouse models. AB - Systemic chemotherapy was considered of modest efficacy in prostate cancer until the recent introduction of taxanes. We took advantage of the known differential effect of camptothecin and docetaxel on human PC-3 and LNCaP prostate cancer cells to determine their effect on sphingosine kinase-1 (SphK1) activity and subsequent ceramide/sphingosine 1-phosphate (S1P) balance in relation with cell survival. In vitro, docetaxel and camptothecin induced strong inhibition of SphK1 and elevation of the ceramide/S1P ratio only in cell lines sensitive to these drugs. SphK1 overexpression in both cell lines impaired the efficacy of chemotherapy by decreasing the ceramide/S1P ratio. Alternatively, silencing SphK1 by RNA interference or pharmacologic inhibition induced apoptosis coupled with ceramide elevation and loss of S1P. The differential effect of both chemotherapeutics was confirmed in an orthotopic PC-3/green fluorescent protein model established in nude mice. Docetaxel induced a stronger SphK1 inhibition and ceramide/S1P ratio elevation than camptothecin. This was accompanied by a smaller tumor volume and the reduced occurrence and number of metastases. SphK1 overexpressing PC-3 cells implanted in animals developed remarkably larger tumors and resistance to docetaxel treatment. These results provide the first in vivo demonstration of SphK1 as a sensor of chemotherapy. PMID- 16357179 TI - Motexafin gadolinium and zinc induce oxidative stress responses and apoptosis in B-cell lymphoma lines. AB - There is an emerging appreciation of the importance of zinc in regulating cancer cell growth and proliferation. Recently, we showed that the anticancer agent motexafin gadolinium (MGd) disrupted zinc metabolism in A549 lung cancer cells, leading, in the presence of exogenous zinc, to cell death. Here, we report the effect of MGd and exogenous zinc on intracellular levels of free zinc, oxidative stress, proliferation, and cell death in exponential phase human B-cell lymphoma and other hematologic cell lines. We find that increased levels of oxidative stress and intracellular free zinc precede and correlate with cell cycle arrest and apoptosis. To better understand the molecular basis of these cellular responses, gene expression profiling analyses were conducted on Ramos cell cultures treated with MGd and/or zinc acetate. Cultures treated with MGd or zinc acetate alone elicited transcriptional responses characterized by induction of metal response element-binding transcription factor-1 (MTF-1)-regulated and hypoxia-inducible transcription factor-1 (HIF-1)-regulated genes. Cultures cotreated with MGd and zinc acetate displayed further increases in the levels of MTF-1- and HIF-1-regulated transcripts as well as additional transcripts regulated by NF-E2-related transcription factor 2. These data provide insights into the molecular changes that accompany the disruption of intracellular zinc homeostasis and support a role for MGd in treatment of B-cell hematologic malignancies. PMID- 16357180 TI - Bcl-2/adenovirus E1B 19 kDa interacting protein-3 knockdown enables growth of breast cancer metastases in the lung, liver, and bone. AB - The mouse breast cancer cell lines 4T1, 4T07, and 67NR are highly tumorigenic but vary in metastatic potential: 4T1 widely disseminates, resulting in secondary tumors in the lung, liver, bone, and brain; 4T07 spreads to the lung and liver but is unable to establish metastatic nodules; 67NR is unable to metastasize. The Bcl-2/adenovirus E1B 19 kDa interacting protein-3 (Bnip-3) was recently shown to be absent after hypoxia in pancreatic cancer cell lines whereas its overexpression restored hypoxia-induced cell death. We found that Bnip-3 expression increased after 6 hours of hypoxia in all cell lines tested but was highest in the nonmetastatic 67NR cells and lowest in the highly metastatic 4T1 cells. Hypoxia-induced expression of Bnip-3 in the disseminating but nonmetastatic 4T07 cells was intermediate compared with 4T1 and 67NR cells. Cleaved caspase-3, a key downstream effector of cell death, increased after 6 hours of hypoxia in the 67NR and 4T07 cells by 1.9- and 2.5-fold, respectively. Conversely, cleaved caspase-3 decreased by 45% in the highly metastatic 4T1 cells after hypoxia. Small interfering RNA oligonucleotides targeting endogenous Bnip-3 blocked cell death and increased clonigenic survival after hypoxic challenge in vitro and increased primary tumor size and enabled metastasis to the lung, liver, and sternum of mice inoculated with 4T07 cells in vivo. These data inversely correlate the hypoxia-induced expression of the cell death protein Bnip-3 to metastatic potential and suggest that loss of Bnip-3 expression is critical for malignant and metastatic evasion of hypoxia-induced cell death. PMID- 16357181 TI - Aberrant transcription from an unrelated promoter can result in MDR-1 expression following drug selection in vitro and in relapsed lymphoma samples. AB - The development of drug resistance in the treatment of cancer remains a major problem. The hallmark of multidrug resistance is cross-resistance to multiple structurally unrelated compounds. The MDR-1 gene encoding P-glycoprotein mediates one of the most extensively studied mechanisms of drug resistance. Previous studies led to the proposal that two promoters control expression of the MDR-1 gene, and these were designated the upstream and downstream promoters. In the present article, we provide evidence that transcripts originating from the putative upstream promoter of MDR-1 are in fact aberrant transcripts whose expression is regulated by nearby genomic sequences that include a human endogenous retroviral long terminal repeat (LTR). Expression of this LTR occurs in all cells. We show that following drug selection, especially in cases where gene amplification has occurred, MDR-1 transcripts can begin near this retroviral LTR with transcription proceeding in the direction opposite of the usual LTR transcription. Because expression of these aberrant MDR-1 transcripts (AMT) is found primarily in drug-resistant cell lines, we conclude that the development of drug resistance or the attendant drug exposure might have a role in the activation of this phenomenon or the selection of cells expressing AMTs. Demonstration of similar aberrant transcripts in tumor samples obtained from patients with relapsed lymphoma suggests that this phenomenon may also occur clinically. PMID- 16357182 TI - Multiple repair pathways mediate tolerance to chemotherapeutic cross-linking agents in vertebrate cells. AB - Cross-linking agents that induce DNA interstrand cross-links (ICL) are widely used in anticancer chemotherapy. Yeast genetic studies show that nucleotide excision repair (NER), Rad6/Rad18-dependent postreplication repair, homologous recombination, and cell cycle checkpoint pathway are involved in ICL repair. To study the contribution of DNA damage response pathways in tolerance to cross linking agents in vertebrates, we made a panel of gene-disrupted clones from chicken DT40 cells, each defective in a particular DNA repair or checkpoint pathway, and measured the sensitivities to cross-linking agents, including cis diamminedichloroplatinum (II) (cisplatin), mitomycin C, and melphalan. We found that cells harboring defects in translesion DNA synthesis (TLS), Fanconi anemia complementation groups (FANC), or homologous recombination displayed marked hypersensitivity to all the cross-linking agents, whereas NER seemed to play only a minor role. This effect of replication-dependent repair pathways is distinctively different from the situation in yeast, where NER seems to play a major role in dealing with ICL. Cells deficient in Rev3, the catalytic subunit of TLS polymerase Polzeta, showed the highest sensitivity to cisplatin followed by fanc-c. Furthermore, epistasis analysis revealed that these two mutants work in the same pathway. Our genetic comprehensive study reveals a critical role for DNA repair pathways that release DNA replication block at ICLs in cellular tolerance to cross-linking agents and could be directly exploited in designing an effective chemotherapy. PMID- 16357183 TI - Immunomodulatory drug lenalidomide (CC-5013, IMiD3) augments anti-CD40 SGN-40 induced cytotoxicity in human multiple myeloma: clinical implications. AB - SGN-40, a humanized immoglobulin G1 (IgG1) anti-CD40 monoclonal antibody, mediates cytotoxicity against human multiple myeloma (MM) cells via suppression of interleukin (IL)-6-induced proliferative and antiapoptotic effects as well as antibody-dependent cell-mediated cytotoxicity (ADCC). Here, we studied the clinical significance of an immunomodulatory drug lenalidomide on SGN-40-induced cytotoxicity against CD138(+)CD40(+) MM lines and patient MM cells. Pretreatment with lenalidomide sensitized MM cells to SGN-40-induced cell death. Combined lenalidomide and SGN-40 significantly induced MM apoptosis, evidenced by enhanced cleavage of caspase-3/8/poly(ADP-ribose)polymerase and increased sub-G(0) cells, compared with either single agent at the same doses. Pretreatment of effector cells with lenalidomide augmented SGN-40-induced MM cell lysis, associated with an increased number of CD56(+)CD3(-) natural killer (NK) cells expressing CD16 and LFA-1. Importantly, pretreatment with lenalidomide or lenalidomide and SGN-40 markedly enhanced NK-cell-mediated lysis of autologous patient MM cells triggered by SGN-40. Lenalidomide also up-regulated CD40L on CD56(+)CD3(-) NK cells, facilitating IL-2-mediated activation of NK cells. In addition, lenalidomide induced the CD56(dim) NK subset, which are more potent mediators of ADCC against target MM cells than the CD56(bright) NK subset. Finally, pretreatment of both effector and target MM cells with lenalidomide markedly enhanced SGN-40-mediated ADCC against CD40-expressing MM cells. These studies, therefore, show that the addition of lenalidomide to SGN-40 enhances cytotoxicity against MM cells, providing the framework for combined lenalidomide and SGN-40 in a new treatment paradigm to both target MM cells directly and induce immune effectors against MM. PMID- 16357184 TI - BGC 945, a novel tumor-selective thymidylate synthase inhibitor targeted to alpha folate receptor-overexpressing tumors. AB - BGC 945 is a cyclopenta[g]quinazoline-based, thymidylate synthase inhibitor specifically transported into alpha-folate receptor (alpha-FR)-overexpressing tumors. Affinity of BGC 945 for the alpha-FR is 70% of the high-affinity ligand folic acid. In contrast to conventional antifolates, BGC 945 has low affinity for the widely expressed reduced-folate carrier (RFC). The K(i) for isolated thymidylate synthase is 1.2 nmol/L and the IC(50) for inhibition of the growth of alpha-FR-negative mouse L1210 or human A431 cells is approximately 7 micromol/L. In contrast, BGC 945 is highly potent in a range of alpha-FR-overexpressing human tumor cell lines (IC(50) approximately 1-300 nmol/L). Pharmacokinetic variables measured following i.v. injection of 100 mg/kg BGC 945 to KB tumor-bearing mice showed rapid plasma clearance (0.021 L/h) and tissue distribution. The terminal half-lives in plasma, liver, kidney, spleen, and tumor were 2, 0.6, 5, 21, and 28 hours, respectively. Tumor BGC 945 concentration at 24 hours was approximately 1 nmol/g tissue, at least 10-fold higher than that in plasma or normal tissues. Inhibition of thymidylate synthase in tissues leads to increased incorporation of 5-[(125)I]-iodo-2'-deoxyuridine ([(125)I]dUrd) into DNA. Forty-eight hours after injection of 100 mg/kg 6RS-BGC 945 ([(125)I]dUrd injected at 24 hours), tumor was the only tissue with incorporation above control level (6-fold). The RFC-mediated thymidylate synthase inhibitor plevitrexed also increased uptake of [(125)I]dUrd in tumor (10-fold) but, in contrast, also caused increased incorporation in other normal tissues such as spleen and small bowel (4.5- and 4.6-fold, respectively). These data suggest that BGC 945 selectively inhibits thymidylate synthase in alpha-FR-overexpressing tumors and should cause minimal toxicity to humans at therapeutic doses. PMID- 16357185 TI - Para to ortho repositioning of the arsenical moiety of the angiogenesis inhibitor 4-(N-(S-glutathionylacetyl)amino)phenylarsenoxide results in a markedly increased cellular accumulation and antiproliferative activity. AB - The synthetic tripeptide arsenical 4-(N-(S-glutathionylacetyl)amino)p phenylarsenoxide (p-GSAO) is an angiogenesis inhibitor that inactivates mitochondrial adenine nucleotide translocase (ANT) by cross-linking a pair of matrix-facing cysteine residues. This causes an increase in superoxide levels and proliferation arrest of endothelial cells followed by mitochondrial depolarization and apoptosis. p-GSAO induces proliferation arrest in endothelial cells and is a selective inhibitor of endothelial cells compared with tumor cells. An analogue of p-GSAO has been made in which the arsenical moiety is at the ortho instead of the para position on the phenyl ring. o-GSAO, like p-GSAO, bound to ANT in a dithiol-dependent manner but was approximately 8-fold more efficient than p-GSAO at triggering the mitochondria permeability transition in isolated mitochondria. o-GSAO was an approximately 50-fold more potent inhibitor of endothelial and tumor cell proliferation than p-GSAO. The mechanism of this effect was a consequence of approximately 300-fold faster rate of accumulation of o-GSAO in the cells, which is due, at least in part, to impaired export by the multidrug resistance-associated protein 1. Administration of o-GSAO to tumor bearing mice delayed tumor growth by inhibiting tumor angiogenesis but there were side effects not observed with p-GSAO administration. PMID- 16357186 TI - Glutathione-related systems and modulation of extracellular signal-regulated kinases are involved in the resistance of AGS adenocarcinoma gastric cells to diallyl disulfide-induced apoptosis. AB - We have previously characterized the cytotoxic action of diallyl disulfide (DADS) on neuroblastoma cells, and we have shown the crucial role of an early and massive reactive oxygen species production in the induction of c-Jun NH(2) terminal kinase-mediated apoptotic pathway. In the present work, we report that DADS is ineffective in inducing apoptosis in a human adenocarcinoma gastric cell line (AGS). In particular, we show that AGS cells are able to recover from the p53/p21-mediated cell cycle arrest in the G(2)-M phase upon DADS treatment without committing cells to death. This event is most likely due to a peculiar surviving pathway of these cells involving: (a) the formation of mixed disulfides between reduced glutathione (GSH) and protein thiols, (b) a higher and inducible glutathione peroxidase activity, and/or (c) an efficient modulation of the phospho-active levels of the extracellular signal-regulated kinases 1 and 2 (ERK 1/2). Moreover, by increasing glutathione peroxidase expression or GSH concentrations, cell cycle arrest is fully abolished; the apoptotic death is induced by either decreasing the availability of intracellular GSH or inhibiting the reactivation of ERK 1/2. Altogether, our data show that ERK 1/2 participates in the active proliferation of AGS cells and that an efficient reactive oxygen species buffering system makes these cells resistant to DADS-mediated detrimental effects. PMID- 16357187 TI - Interleukin-13-regulated M2 macrophages in combination with myeloid suppressor cells block immune surveillance against metastasis. AB - CD1-deficient mice reject established, disseminated 4T1 metastatic mammary cancer and survive indefinitely if their primary mammary tumors are surgically removed. This highly effective immune surveillance is due to three interacting mechanisms: (a) the generation of inducible nitric oxide synthase (iNOS)-producing M1 macrophages that are tumoricidal for 4T1 tumor cells; (b) a rapid decrease in myeloid-derived Gr1(+)CD11b(+) suppressor cells that are elevated and down regulate the CD3zeta chain when primary tumor is present and that suppress T cells by producing arginase; and (c) production of activated lymphocytes. Macrophages from wild-type BALB/c mice are polarized by interleukin-13 (IL-13) towards a tumor-promoting M2 phenotype, thereby inhibiting the generation of tumoricidal M1 macrophages. In contrast, CD1(-/-) mice, which are deficient for IL-13 because they lack IL-13-producting NKT cells, generate M1 macrophages that are cytotoxic for 4T1 via the production of nitric oxide. Although tumoricidal macrophages are a necessary component of immune surveillance in CD1(-/-) mice, they alone are not sufficient for tumor resistance because IL-4Ralpha(-/-) mice have M1 macrophages and retain high levels of myeloid suppressor cells after surgery; in addition, they are susceptible to 4T1 metastatic disease. These results show that effective immune surveillance against established metastatic disease is negatively regulated by IL-13 and requires the induction of tumoricidal M1 macrophages and lymphocytes combined with a reduction in tumor induced myeloid suppressor cells. PMID- 16357188 TI - Activation of tumor-associated macrophages by the vascular disrupting agent 5,6 dimethylxanthenone-4-acetic acid induces an effective CD8+ T-cell-mediated antitumor immune response in murine models of lung cancer and mesothelioma. AB - 5,6-Dimethylxanthenone-4-acetic acid (DMXAA) is a small molecule in the flavanoid class that has antitumor activity thought to be due to ability to induce high local levels of tumor necrosis factor (TNF)-alpha that disrupt established blood vessels within tumors. The drug has completed phase 1 testing in humans and is currently in phase 2 trials in combination with chemotherapy. Although characterized as a "vascular disrupting agent," there are some studies suggesting that DMXAA also has effects on the immune system that are important for its efficacy. The goal of this study was to carefully define the immune effects of DMXAA in a series of murine lung cancer and mesothelioma cell lines with varying immunologic characteristics. We show that DMXAA efficiently activated tumor associated macrophages to release a variety of immunostimulatory cytokines and chemokines, including TNF-alpha; IFN-inducible protein-10; interleukin-6; macrophage inflammatory protein-2; monocyte chemotactic protein-1; and regulated on activation, normal T-cell expressed, and secreted. DMXAA treatment was highly effective in both small and large flank tumors. Animals cured of tumors by DMXAA generated a systemic memory response and were resistant to tumor cell rechallenge. DMXAA treatment led to initial tumor infiltration with macrophages that was followed by an influx of CD8(+) T cells. These CD8(+) T cells were required for antitumor efficacy because tumor inhibitory activity was lost in nude mice, mice depleted of CD8(+) T cells, and perforin knockout mice, but not in CD4(+) T-cell-depleted mice. These data show that activation of tumor associated macrophages by DMXAA is an efficient way to generate a CD8(+) T-cell dependent antitumor immune response even in animals with relatively nonimmunogenic tumors. Given these properties, DMXAA might also be useful in boosting other forms of immunotherapy. PMID- 16357189 TI - Coactivation of liver receptor homologue-1 by peroxisome proliferator-activated receptor gamma coactivator-1alpha on aromatase promoter II and its inhibition by activated retinoid X receptor suggest a novel target for breast-specific antiestrogen therapy. AB - Aromatase inhibitors target the production of estrogen in breast adipose tissue, but in doing so, also decrease estrogen formation in bone and other sites, giving rise to deleterious side effects, such as bone loss and arthralgia. Thus, it would be clinically useful to selectively inhibit aromatase production in breast. In this regard, we have determined that the orphan nuclear receptor liver receptor homologue-1 (LRH-1) is a specific transcriptional activator of aromatase gene expression in human breast preadipocytes but not in other tissues of postmenopausal women. In this study, we show that the coactivator peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) is a physiologically relevant modulator of LRH-1, and that its transcriptional activity can be inhibited effectively using receptor-interacting peptide antagonists that prevent PGC-1alpha recruitment. Interestingly, we note that all of these peptides also interact in an agonist-dependent manner with retinoid X receptor alpha (RXRalpha), suggesting that these two receptors may compete for limiting cofactors within target cells. In support of this hypothesis, we show that 9-cis-retinoic acid, acting through RXR, inhibits both the basal and PGC 1alpha-induced transcriptional activity of LRH-1. The importance of this finding was confirmed by showing that LRH-1-dependent, PGC-1alpha-stimulated regulation of aromatase gene expression in primary human breast preadipocytes was effectively suppressed by RXR agonists. We infer from these data that LRH-1 is a bona fide target whose inhibition would selectively block aromatase expression in breast, while sparing other sites of expression. PMID- 16357190 TI - Sequence variants of Toll-like receptor 4 and susceptibility to prostate cancer. AB - Chronic inflammation has been hypothesized to be a risk factor for prostate cancer. The Toll-like receptor 4 (TLR4) presents the bacterial lipopolysaccharide (LPS), which interacts with ligand-binding protein and CD14 (LPS receptor) and activates expression of inflammatory genes through nuclear factor-kappaB and mitogen-activated protein kinase signaling. A previous case-control study found a modest association of a polymorphism in the TLR4 gene [11381G/C, GG versus GC/CC: odds ratio (OR), 1.26] with risk of prostate cancer. We assessed if sequence variants of TLR4 were associated with the risk of prostate cancer. In a nested case-control design within the Health Professionals Follow-up Study, we identified 700 participants with prostate cancer diagnosed after they had provided a blood specimen in 1993 and before January 2000. Controls were 700 age matched men without prostate cancer who had had a prostate-specific antigen test after providing a blood specimen. We genotyped 16 common (>5%) single nucleotide polymorphisms (SNP) discovered in a resequencing study spanning TLR4 to test for association between sequence variation in TLR4 and prostate cancer. Homozygosity for the variant alleles of eight SNPs was associated with a statistically significantly lower risk of prostate cancer (TLR4_1893, TLR4_2032, TLR4_2437, TLR4_7764, TLR4_11912, TLR4_16649, TLR4_17050, and TLR4_17923), but the TLR4_15844 polymorphism corresponding to 11381G/C was not associated with prostate cancer (GG versus CG/CC: OR, 1.01; 95% confidence interval, 0.79-1.29). Six common haplotypes (cumulative frequency, 81%) were observed; the global test for association between haplotypes and prostate cancer was statistically significant (chi(2) = 14.8 on 6 degrees of freedom; P = 0.02). Two common haplotypes were statistically significantly associated with altered risk of prostate cancer. Inherited polymorphisms of the innate immune gene TLR4 are associated with risk of prostate cancer. PMID- 16357191 TI - A prospective study of meat and meat mutagens and prostate cancer risk. AB - High-temperature cooked meat contains heterocyclic amines, including 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and polycyclic aromatic hydrocarbons, such as benzo(a)pyrene (BaP). In rodents, a high intake of PhIP induces prostate tumors. We prospectively investigated the association between meat and meat mutagens, specifically PhIP, and prostate cancer risk in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Diet was assessed using a 137-item food frequency questionnaire and a detailed meat-cooking questionnaire linked to a database for BaP and the heterocyclic amines 2-amino 3,8-dimethylimidazo[4,5-b]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5 f]quinoxaline (DiMeIQx), and PhIP. During follow-up, we ascertained a total of 1,338 prostate cancer cases among 29,361 men; of these, 868 were incident cases (diagnosed after the first year of follow-up) and 520 were advanced cases (stage III or IV or a Gleason score of > or =7). Total, red, or white meat intake was not associated with prostate cancer risk. More than 10 g/d of very well done meat, compared with no consumption, was associated with a 1.4-fold increased risk of prostate cancer [95% confidence interval (95% CI), 1.05-1.92] and a 1.7-fold increased risk (95% CI, 1.19-2.40) of incident disease. Although there was no association with MeIQx and DiMeIQx, the highest quintile of PhIP was associated with a 1.2-fold increased risk of prostate cancer (95% CI, 1.01-1.48) and a 1.3 fold increased risk of incident disease (95% CI, 1.01-1.61). In conclusion, very well done meat was positively associated with prostate cancer risk. In addition, this study lends epidemiologic support to the animal studies, which have implicated PhIP as a prostate carcinogen. PMID- 16357192 TI - Stereotypic and random patterns of connectivity in the larval mushroom body calyx of Drosophila. AB - The larval brain of Drosophila is a useful model to study olfactory processing because of its cellular simplicity. The early stages of central olfactory processing involve the detection of odor features, but the coding mechanisms that transform them into a representation in higher brain centers is not clear. Here we examine the pattern of connectivity of the main neurons that process olfactory information in the calyx (dendritic region) of the mushroom bodies, a higher brain center essential for associative olfactory learning. The larval calyx has a glomerular organization. We generated a map of calyx glomeruli, using both anatomical criteria and the pattern of innervation by subsets of its input neurons (projection neurons), molecularly identified by GAL4 markers. Thus, we show that projection neurons innervate calyx glomeruli in a stereotypic manner. By contrast, subsets of mushroom body neurons (Kenyon cells) that are labeled by GAL4 markers show no clear preference for specific glomeruli. Clonal subsets of Kenyon cells show some preference for subregions of the calyx, implying that they receive distinct input. However, at the level of individual glomeruli, dendritic terminals of larval-born Kenyon cells innervate about six glomeruli, apparently randomly. These results are consistent with a model in which Kenyon cells process olfactory information by integrating different inputs from several calyx glomeruli in a combinatorial manner. PMID- 16357193 TI - A selection for mutants that interfere with folding of Escherichia coli thioredoxin-1 in vivo. AB - Escherichia coli thioredoxin is normally a cytoplasmic protein involved in the reduction of disulfide bonds. However, thioredoxin can be translocated to the periplasm when it is attached to a cotranslational signal sequence. When exported to the periplasm, it can partially replace the activity of DsbA in promoting the formation of disulfide bonds. In contrast, when thioredoxin is fused to a posttranslational signal sequence, very little of it appears in the periplasm. We propose that this absence of posttranslational export is due to the rapid folding of thioredoxin in the cytoplasm. We sought mutants of thioredoxin that retarded its folding in the cytoplasm, which we accomplished by fusing thioredoxin to a posttranslational signal sequence and selecting for mutants in which thioredoxin was exported to the periplasm, where it could replace DsbA. The collection of mutants obtained represents a limited number of amino acid changes in the protein. In vitro studies on purified mutant proteins show that all but one are defective in the kinetics and thermodynamics of protein folding. We propose that the slower folding of the thioredoxin mutant proteins in the cytoplasm allows their export by a posttranslational pathway. We discuss some implications of this class of mutants for aspects of the folding pathway of thioredoxin and for its mechanism of export. In particular, the finding that a folding mutant that allows protein translocation alters an amino acid at the C terminus of the protein suggests that the degree to which thioredoxin folds during its translation must be severely restricted. PMID- 16357194 TI - Processes conducive to the release and transport of arsenic into aquifers of Bangladesh. AB - Arsenic is a contaminant in the groundwater of Holocene aquifers in Bangladesh, where approximately 57 million people drink water with arsenic levels exceeding the limits set by the World Health Organization. Although arsenic is native to the sediments, the means by which it is released to groundwater remains unresolved. Contrary to the current paradigm, ferric (hydr)oxides appear to dominate the partitioning of arsenic in the near surface but have a limited impact at aquifer depths where wells extract groundwater with high arsenic concentrations. We present a sequence of evidence that, taken together, suggest that arsenic may be released in the near surface and then transported to depth. We establish that (i) the only portion of the sediment profile with conditions destabilizing to arsenic in our analysis is in the surface or near-surface environment; (ii) a consistent input of arsenic via sediment deposition exists; (iii) retardation of arsenic transport is limited in the aquifers; and (iv) groundwater recharge occurs at a rate sufficient to necessitate continued input of arsenic to maintain observed concentrations. Our analyses thus lead to the premise that arsenic is liberated in surface and near-surface sediments through cyclic redox conditions and is subsequently transported to well depth. Influx of sediment and redox cycling provide a long-term source of arsenic that when liberated in the near surface is only weakly partitioned onto sediments deeper in the profile and is transported through aquifers by groundwater recharge. PMID- 16357195 TI - MicroRNA1 influences cardiac differentiation in Drosophila and regulates Notch signaling. AB - MicroRNAs (miRNAs) are genomically encoded small RNAs that hybridize with messenger RNAs, resulting in degradation or translational inhibition of targeted transcripts. The potential for miRNAs to regulate cell-lineage determination or differentiation from pluripotent progenitor or stem cells is unknown. Here, we show that microRNA1 (miR-1) is an ancient muscle-specific gene conserved in sequence and expression in Drosophila. Drosophila miR-1 (dmiR-1) is regulated through a serum response factor-like binding site in cardiac progenitor cells. Loss- and gain-of-function studies demonstrated a role for dmiR-1 in modulating cardiogenesis and in maintenance of muscle-gene expression. We provide in vivo evidence that dmiR-1 targets transcripts encoding the Notch ligand Delta, providing a potential mechanism for the expansion of cardiac and muscle progenitor cells and failure of progenitor cell differentiation in some dmiR-1 mutants. These findings demonstrate that dmiR-1 may "fine-tune" critical steps involved in differentiation of cardiac and somatic muscle progenitors and targets a pathway required for progenitor cell specification and asymmetric cell division. PMID- 16357196 TI - Endocannabinoids regulate interneuron migration and morphogenesis by transactivating the TrkB receptor. AB - In utero exposure to Delta(9)-tetrahydrocannabinol (Delta(9)-THC), the active component from marijuana, induces cognitive deficits enduring into adulthood. Although changes in synaptic structure and plasticity may underlie Delta(9)-THC induced cognitive impairments, the neuronal basis of Delta(9)-THC-related developmental deficits remains unknown. Using a Boyden chamber assay, we show that agonist stimulation of the CB(1) cannabinoid receptor (CB(1)R) on cholecystokinin-expressing interneurons induces chemotaxis that is additive with brain-derived neurotrophic factor (BDNF)-induced interneuron migration. We find that Src kinase-dependent TrkB receptor transactivation mediates endocannabinoid (eCB)-induced chemotaxis in the absence of BDNF. Simultaneously, eCBs suppress the BDNF-dependent morphogenesis of interneurons, and this suppression is abolished by Src kinase inhibition in vitro. Because sustained prenatal Delta(9) THC stimulation of CB(1)Rs selectively increases the density of cholecystokinin expressing interneurons in the hippocampus in vivo, we conclude that prenatal CB(1)R activity governs proper interneuron placement and integration during corticogenesis. Moreover, eCBs use TrkB receptor-dependent signaling pathways to regulate subtype-selective interneuron migration and specification. PMID- 16357197 TI - Analysis and mapping of randomly chosen bacterial artificial chromosome clones from hexaploid bread wheat. AB - The current view of wheat genome composition is that genes are compartmentalized into gene-rich and gene-poor regions. This model can be tested by analyzing randomly selected bacterial artificial chromosome (BAC) clones for gene content, followed by placement of these BACs onto physical and genetic maps. Map localization could be difficult for BACs that consist entirely of repeated elements. We therefore developed a technique where repeat junctions are used to generate unique markers. Four BAC clones from hexaploid wheat variety Chinese Spring were randomly selected and sequenced at 4- to 6-fold redundancy. About 50% of the BAC sequences corresponded to previously identified repeats, mainly LTR retrotransposons, whereas most of the remaining DNA consisted of sequences with unknown origin or function. The average gene content was <1%, although each BAC contained one or two identified genes. Repeat boundaries were amplified and used to map each clone to a chromosome arm. Extrapolation from wheat-rice comparative knowledge suggests that three of the four BAC clones originate from "gene-rich" regions of the wheat genome. Nevertheless, because these BACs carry only a single gene (two BACs) or two genes (one BAC), the predicted gene density is approximately 1 gene per 75 kb, which is considerably lower than previously estimated gene densities (one gene per 5-20 kb) for gene-rich regions in wheat. This analysis of randomly selected wheat BAC clones suggests that genes are more evenly distributed in wheat than previously believed and substantiates the need for large-scale random BAC sequencing to determine wheat genome organization. PMID- 16357198 TI - How to make a curved Drosophila bristle using straight actin bundles. AB - This, our Inaugural Article as Academy Members, is ironically our swan song from the field of the actin cytoskeleton. By reviewing what we have learned and what we think is going on during development, we hope to lure you, the reader, into applying your skills to the bristle cell. The processes of the assembly and disassembly of actin bundles is laid out in time and space in an organism that lends itself to genetic manipulation. The cell provides every process you could want: filament nucleation, growth of microvilli, joining of microvillar bundles into modules, assembly of modules into bundles, time-dependent use of at least two crossbridging proteins, filament turnover, treadmilling, disassembly, and filament translocation. PMID- 16357199 TI - Grammatical Subjects in home sign: Abstract linguistic structure in adult primary gesture systems without linguistic input. AB - Language ordinarily emerges in young children as a consequence of both linguistic experience (for example, exposure to a spoken or signed language) and innate abilities (for example, the ability to acquire certain types of language patterns). One way to discern which aspects of language acquisition are controlled by experience and which arise from innate factors is to remove or manipulate linguistic input. However, experimental manipulations that involve depriving a child of language input are impossible. The present work examines the communication systems resulting from natural situations of language deprivation and thus explores the inherent tendency of humans to build communication systems of particular kinds, without any conventional linguistic input. We examined the gesture systems that three isolated deaf Nicaraguans (ages 14-23 years) have developed for use with their hearing families. These deaf individuals have had no contact with any conventional language, spoken or signed. To communicate with their families, they have each developed a gestural communication system within the home called "home sign." Our analysis focused on whether these systems show evidence of the grammatical category of Subject. Subjects are widely considered to be universal to human languages. Using specially designed elicitation tasks, we show that home signers also demonstrate the universal characteristics of Subjects in their gesture productions, despite the fact that their communicative systems have developed without exposure to a conventional language. These findings indicate that abstract linguistic structure, particularly the grammatical category of Subject, can emerge in the gestural modality without linguistic input. PMID- 16357200 TI - A therapeutic aptamer inhibits angiogenesis by specifically targeting the heparin binding domain of VEGF165. AB - Aptamers recognize their targets with extraordinary affinity and specificity. The aptamer-based therapeutic, Macugen, is derived from a modified 2'fluoro pyrimidine RNA inhibitor to vascular endothelial growth factor (VEGF) and is now being used to treat the wet form of age-related macular degeneration. This VEGF(165) aptamer binds specifically to the VEGF(165) isoform, a dimeric protein with a receptor-binding domain and a heparin-binding domain (HBD). To understand the molecular recognition between VEGF and this aptamer, binding experiments were used to show that the HBD contributes the majority of binding energy in the VEGF(165)-aptamer complex. A tissue culture-based competition assay demonstrated that the HBD effectively competes with VEGF(165) for aptamer binding in vivo. Comparison of NMR spectra revealed that structural features of the smaller HBD aptamer complex are present in the full-length VEGF(164)-aptamer complex. These data show that the HBD provides the binding site for the aptamer and is the primary determinant for the affinity and specificity in the VEGF(165)-aptamer complex. PMID- 16357201 TI - Jasmonate-inducible plant enzymes degrade essential amino acids in the herbivore midgut. AB - The plant hormone jasmonic acid (JA) activates host defense responses against a broad spectrum of herbivores. Although it is well established that JA controls the expression of a large set of target genes in response to tissue damage, very few gene products have been shown to play a direct role in reducing herbivore performance. To test the hypothesis that JA-inducible proteins (JIPs) thwart attack by disrupting digestive processes in the insect gut, we used a MS-based approach to identify host proteins that accumulate in the midgut of Manduca sexta larvae reared on tomato (Solanum lycopersicum) plants. We show that two JIPs, arginase and threonine deaminase (TD), act in the M. sexta midgut to catabolize the essential amino acids Arg and Thr, respectively. Transgenic plants that overexpress arginase were more resistant to M. sexta larvae, and this effect was correlated with reduced levels of midgut Arg. We present evidence indicating that the ability of TD to degrade Thr in the midgut is enhanced by herbivore-induced proteolytic removal of the enzyme's C-terminal regulatory domain, which confers negative feedback regulation by isoleucine in planta. Our results demonstrate that the JA signaling pathway strongly influences the midgut protein content of phytophagous insects and support the hypothesis that catabolism of amino acids in the insect digestive tract by host enzymes plays a role in plant protection against herbivores. PMID- 16357202 TI - Ribosome exit tunnel can entropically stabilize alpha-helices. AB - Several experiments have suggested that newly synthesized polypeptide chains can adopt helical structures deep within the ribosome exit tunnel. We hypothesize that confinement in the roughly cylindrical tunnel can entropically stabilize alpha-helices. The hypothesis is validated by using theory and simulations of coarse-grained off-lattice models. The model helix, which is unstable in the bulk, is stabilized in a cylindrical cavity provided the diameter (D) of the cylinder exceeds a critical value D*. When D < D* both the helical content and the helix-coil transition temperature (T(f)) decrease abruptly. Surprisingly, we find that the stability of the alpha-helix depends on the number (N) of amino acid residues. Entropic stabilization, as measured by changes in T(f), increases nonlinearly as N increases. The simulation results are in quantitative agreement with a standard helix-coil theory that takes into account entropy cost of confining a polypeptide chain in a cylinder. The results of this work are in qualitative accord with most of the findings of a recent experiment in which N dependent ribosome-induced helix stabilization of transmembrane sequences was measured by fluorescence resonance energy transfer. PMID- 16357203 TI - Role for hypocretin in mediating stress-induced reinstatement of cocaine-seeking behavior. AB - Hypocretin-1 and -2 (Hcrt-1 and Hcrt-2), also referred to as orexin-A and -B, are neuropeptides synthesized by a few thousand neurons in the lateral hypothalamus. Hypocretin-containing neurons project throughout the brain, with a prominent input to basal forebrain structures involved in motivation, reward, and stress. However, the role of hypocretins in addiction-related behaviors remains largely unexplored. Here we show that intracerebroventricular infusions of Hcrt-1 lead to a dose-related reinstatement of cocaine seeking without altering cocaine intake in rats. Hcrt-1 also dramatically elevates intracranial self-stimulation thresholds, indicating that, unlike treatments with reinforcing properties such as cocaine, Hcrt-1 negatively regulates the activity of brain reward circuitries. Hypocretin-induced reinstatement of cocaine seeking was prevented by blockade of noradrenergic and corticotropin-releasing factor systems, suggesting that Hcrt-1 reinstated drug seeking through induction of a stress-like state. Consistent with this interpretation, the selective Hcrt-1 receptor antagonist SB-334867 blocked footshock-induced reinstatement of previously extinguished cocaine-seeking behavior. These findings reveal a previously unidentified role for hypocretins in driving drug seeking through activation of stress pathways in the brain. PMID- 16357204 TI - Structure of an archaeal virus capsid protein reveals a common ancestry to eukaryotic and bacterial viruses. AB - Archaea and their viruses are poorly understood when compared with the Eukarya and Bacteria domains of life. We report here the crystal structure of the major capsid protein (MCP) of the Sulfolobus turreted icosahedral virus, an archaeal virus isolated from an acidic hot spring (pH 2-4, 72-92 degrees C) in Yellowstone National Park. The structure is nearly identical to the MCP structures of the eukaryotic Paramecium bursaria Chlorella virus, and the bacteriophage PRD1, and shows a common fold with the mammalian adenovirus. Structural analysis of the capsid architecture, determined by fitting the subunit into the electron cryomicroscopy reconstruction of the virus, identified a number of key interactions that are akin to those observed in adenovirus and PRD1. The similar capsid proteins and capsid architectures strongly suggest that these viral capsids originated and evolved from a common ancestor. Hence, this work provides a previously undescribed example of a viral relationship spanning the three domains of life (Eukarya, Bacteria, and Archaea). The MCP structure also provides insights into the stabilizing forces required for extracellular hyperthermophilic proteins to tolerate high-temperature hot springs. PMID- 16357205 TI - Embryonic stem cell-derived hematopoietic stem cells. AB - Despite two decades of studies documenting the in vitro blood-forming potential of murine embryonic stem cells (ESCs), achieving stable long-term blood engraftment of ESC-derived hematopoietic stem cells in irradiated mice has proven difficult. We have exploited the Cdx-Hox pathway, a genetic program important for blood development, to enhance the differentiation of ESCs along the hematopoietic lineage. Using an embryonic stem cell line engineered with tetracycline-inducible Cdx4, we demonstrate that ectopic Cdx4 expression promotes hematopoietic mesoderm specification, increases hematopoietic progenitor formation, and, together with HoxB4, enhances multilineage hematopoietic engraftment of lethally irradiated adult mice. Clonal analysis of retroviral integration sites confirms a common stem cell origin of lymphoid and myeloid populations in engrafted primary and secondary mice. These data document the cardinal stem cell features of self renewal and multilineage differentiation of ESC-derived hematopoietic stem cells. PMID- 16357206 TI - The global transcriptional regulatory network for metabolism in Escherichia coli exhibits few dominant functional states. AB - A principal aim of systems biology is to develop in silico models of whole cells or cellular processes that explain and predict observable cellular phenotypes. Here, we use a model of a genome-scale reconstruction of the integrated metabolic and transcriptional regulatory networks for Escherichia coli, composed of 1,010 gene products, to assess the properties of all functional states computed in 15,580 different growth environments. The set of all functional states of the integrated network exhibits a discernable structure that can be visualized in 3 dimensional space, showing that the transcriptional regulatory network governing metabolism in E. coli responds primarily to the available electron acceptor and the presence of glucose as the carbon source. This result is consistent with recently published experimental data. The observation that a complex network composed of 1,010 genes is organized to achieve few dominant modes demonstrates the utility of the systems approach for consolidating large amounts of genome scale molecular information about a genome and its regulation to elucidate an organism's preferred environments and functional capabilities. PMID- 16357207 TI - Na+ channel-mediated Ca2+ entry leads to glutamate secretion in mouse neocortical preplate. AB - Before synaptogenesis, early excitability implicating voltage-dependent and transmitter-activated channels is known to be crucial for neuronal development. We previously showed that preplate (PP) neurons of the mouse neocortex express functional Na(+) channels as early as embryonic day 12. In this study, we investigated the role of these Na(+) channels in signaling during early development. In the neocortex of embryonic-day-13 mice, activation of Na(+) channels with veratridine induced a large Ca(2+) response throughout the neocortex, even in cell populations that lack the Na(+) channel. This Na(+) dependent Ca(2+) activity requires external Ca(2+) and is completely blocked by inhibitors of Na(+)/Ca(2+) exchangers. Moreover, veratridine-induced Ca(2+) increase coincides with a burst of exocytosis in the PP. In parallel, we show that Na(+) channel stimulation enhances glutamate secretion in the neocortical wall. Released glutamate triggers further Ca(2+) response in PP and ventricular zone, as indicated by the decreased response to veratridine in the presence of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor and NMDA receptor inhibitors. Therefore, the combined activation of the Na(+) channel and the Na(+)/Ca(2+) exchanger triggers Ca(2+) signaling in the PP neurons, leading to glutamate secretion, which amplifies the signal and serves as an autocrine/paracrine transmitter before functional synapses are formed in the neocortex. Membrane depolarization induced by glycine receptors activation could be one physiological activator of this Na(+) channel-dependent pathway. PMID- 16357208 TI - Maintenance of presynaptic function by AMPA receptor-mediated excitatory postsynaptic activity in adult brain. AB - Activity-dependent synaptic modification occurs in both developing and mature animals. For reliable information transfer and storage, however, once established, synapses must be maintained stably. We investigated how chronic blockade of neuronal activity or alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)-type glutamate receptors affects excitatory climbing fiber (CF) to Purkinje cell (PC) synapses in adult mouse cerebellum. Both treatments caused reduced glutamate concentration transient at the synaptic cleft, decreased frequency of quantal excitatory postsynaptic current, and diminished CF innervation of PC shaft dendrites but no change in CF's release probability. These results indicate that, in the mature cerebellum, AMPA receptor mediated excitatory postsynaptic activity maintains CF's functional glutamate release sites and its innervation of PC shaft dendrites. PMID- 16357209 TI - Interaction between the dihydropyridine receptor Ca2+ channel beta-subunit and ryanodine receptor type 1 strengthens excitation-contraction coupling. AB - Previous studies have shown that the skeletal dihydropyridine receptor (DHPR) pore subunit Ca(V)1.1 (alpha1S) physically interacts with ryanodine receptor type 1 (RyR1), and a molecular signal is transmitted from alpha1S to RyR1 to trigger excitation-contraction (EC) coupling. We show that the beta-subunit of the skeletal DHPR also binds RyR1 and participates in this signaling process. A novel binding site for the DHPR beta1a-subunit was mapped to the M(3201) to W(3661) region of RyR1. In vitro binding experiments showed that the strength of the interaction is controlled by K(3495)KKRR_ _R(3502), a cluster of positively charged residues. Phenotypic expression of skeletal-type EC coupling by RyR1 with mutations in the K(3495)KKRR_ _R(3502) cluster was evaluated in dyspedic myotubes. The results indicated that charge neutralization or deletion severely depressed the magnitude of RyR1-mediated Ca(2+) transients coupled to voltage dependent activation of the DHPR. Meantime the Ca(2+) content of the sarcoplasmic reticulum was not affected, and the amplitude and activation kinetics of the DHPR Ca(2+) currents were slightly affected. The data show that the DHPR beta-subunit, like alpha1S, interacts directly with RyR1 and is critical for the generation of high-speed Ca(2+) signals coupled to membrane depolarization. These findings indicate that EC coupling in skeletal muscle involves the interplay of at least two subunits of the DHPR, namely alpha1S and beta1a, interacting with possibly different domains of RyR1. PMID- 16357210 TI - Cell death mechanism and protective effect of erythropoietin after focal ischemia in the whisker-barrel cortex of neonatal rats. AB - Cell death induced by the combined insult of hypoxia-ischemia in neonatal rodents has been extensively investigated. Ischemia-only-induced cell death, however, has been much less characterized. Based on the notion that 1) ischemic stroke is a relatively common disorder in human neonates, and 2) developing cells are more susceptible to apoptosis, the present study examined whether typical apoptosis was induced by cerebral ischemia in a new neonatal rat model. Erythropoietin (EPO; Epoetin) was tested for its protective effect against ischemia-induced cell death. Postnatal day 7 rats were subjected to permanent occlusion of the middle cerebral artery branch supplying the right whisker-barrel cortex. Terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeled-positive cells in the ischemic region were detectable 4 h after ischemia and reached a peak level 16 h later. The cell death was preceded by caspase activation and cytochrome c release. Cell body shrinkage was evident among damaged cells. Agarose gel electrophoresis showed DNA damage with a smear pattern as well as DNA laddering. Electron microscopy demonstrated apoptotic features such as cell shrinkage, chromatin condensation, and fragmentation; meanwhile, necrotic alterations coexisted in the cytoplasm. EPO treatment increased signal transducers and activators of transcription-5 and Bcl-2 levels, markedly attenuated apoptotic cell death, and reduced ischemic infarct in the cortex. It is suggested that focal ischemia in the developing brain causes cell death with prominent apoptotic features coexisting with some characteristics of necrosis. This is consistent with the concept of hybrid death described previously in cultures and adult or developing brain. EPO may be explored as a potential therapy for neonatal ischemic stroke. PMID- 16357211 TI - Tectonic, a novel regulator of the Hedgehog pathway required for both activation and inhibition. AB - We report the identification of a novel protein that participates in Hedgehog mediated patterning of the neural tube. This protein, named Tectonic, is the founding member of a previously undescribed family of evolutionarily conserved secreted and transmembrane proteins. During neural tube development, mouse Tectonic is required for formation of the most ventral cell types and for full Hedgehog (Hh) pathway activation. Epistasis analyses reveal that Tectonic modulates Hh signal transduction downstream of Smoothened (Smo) and Rab23. Interestingly, characterization of Tectonic Shh and Tectonic Smo double mutants indicates that Tectonic plays an additional role in repressing Hh pathway activity. PMID- 16357212 TI - Two classes of endogenous small RNAs in Tetrahymena thermophila. AB - Endogenous small RNAs function in RNA interference (RNAi) pathways to guide RNA cleavage, translational repression, or methylation of DNA or chromatin. In Tetrahymena thermophila, developmentally regulated DNA elimination is governed by an RNAi mechanism involving approximately 27-30-nucleotide (nt) RNAs. Here we characterize the sequence features of the approximately 27-30-nt RNAs and a approximately 23-24-nt RNA class representing a second RNAi pathway. The approximately 23-24-nt RNAs accumulate strain-specifically manner and map to the genome in clusters that are antisense to predicted genes. These findings reveal the existence of distinct endogenous RNAi pathways in the unicellular T. thermophila, a complexity previously demonstrated only in multicellular organisms. PMID- 16357213 TI - The Fanconi Anemia/BRCA pathway: new faces in the crowd. AB - Over the past few years, study of the rare inherited chromosome instability disorder, Fanconi Anemia (FA), has uncovered a novel DNA damage response pathway. Through the cooperation of multiple proteins, this pathway regulates a complicated cellular response to DNA cross-linking agents and other genotoxic stresses. In this article we review recent data identifying new components of the FA pathway that implicate it in several aspects of the DNA damage response, including the direct processing of DNA, translesion synthesis, homologous recombination, and cell cycle regulation. We also discuss new findings that explain how the FA pathway is regulated through the processes of ubiquitination and deubiquitination. We then consider the clinical implications of our current understanding of the FA pathway, particularly in the development and treatment of malignancy in heterozygous carriers of FA mutations or in patients with sporadic cancers. We consider how recent studies of p53-mediated apoptosis and loss of p53 function in models of FA may help explain the clinical features of the disease and finally present a hypothesis to account for the specificity of the FA pathway in the response to DNA cross-links. PMID- 16357214 TI - Polycomb mediates Myc autorepression and its transcriptional control of many loci in Drosophila. AB - Aberrant accumulation of the Myc oncoprotein propels proliferation and induces carcinogenesis. In normal cells, however, an abundance of Myc protein represses transcription at the c-myc locus. Cancer cells often lose this autorepression. We examined the control of myc in Drosophila and show here that the Drosophila ortholog, dmyc, also undergoes autorepression. We find that the developmental repressor Polycomb (Pc) is required for dmyc autorepression, and that this Pc dMyc-mediated repression spreads across an 875-kb region encompassing the dmyc gene. To further investigate the relationship between Myc and Polycomb, we used microarrays to identify genes regulated by each, and identify a striking relationship between the two: A large set of dMyc activation targets is normally repressed by Pc, and 73% of dMyc repression targets require Pc for this repression. Chromatin immunoprecipitation confirmed that many dMyc-Pc-repressed loci have an epigenetic mark recognized by Pc. Our results suggest a novel relationship between Myc and Polycomb, wherein Myc enhances Polycomb repression in order to repress targets, and Myc suppresses Polycomb repression in order to activate targets. PMID- 16357215 TI - The Drosophila microRNA iab-4 causes a dominant homeotic transformation of halteres to wings. AB - The Drosophila Bithorax Complex encodes three well-characterized homeodomain proteins that direct segment identity, as well as several noncoding RNAs of unknown function. Here, we analyze the iab-4 locus, which produces the microRNAs iab-4-5p and iab-4-3p. iab-4 is analogous to miR-196 in vertebrate Hox clusters. Previous studies demonstrate that miR-196 interacts with the Hoxb8 3' untranslated region. Evidence is presented that miR-iab-4-5p directly inhibits Ubx activity in vivo. Ectopic expression of mir-iab-4-5p attenuates endogenous Ubx protein accumulation and induces a classical homeotic mutant phenotype: the transformation of halteres into wings. These findings provide the first evidence for a noncoding homeotic gene and raise the possibility that other such genes occur within the Bithorax complex. We also discuss the regulation of mir-iab-4 expression during development. PMID- 16357216 TI - A human, ATP-independent, RISC assembly machine fueled by pre-miRNA. AB - RNA interference (RNAi) is mediated by RNA-induced silencing complexes (RISCs), which are guided by microRNAs (miRNAs) or short interfering RNAs (siRNAs) to cognate RNA targets. In humans, the catalytic engine of RISC is a ribonucleoprotein formed by the Argonaute2 (Ago2) protein and either miRNA (miRNP) or siRNA (siRNP). The Dicer nuclease produces mature miRNAs and siRNAs from pre-miRNAs and double-stranded RNA (dsRNA), respectively, and associates with Ago2. Here, we studied the assembly of human RISC by presenting pre-miRNA to immunopurified complexes that contain Ago2, Dicer, and TRBP. Mature miRNAs were produced in an ATP-independent manner and guided specific cleavage of cognate RNA targets in a pattern that is typical of RISC. This de novo formed RISC activity dissociated from Dicer. The asymmetry of the RISC loading process was fully recapitulated in this system, which, however, could not efficiently assemble RISC from siRNA duplexes. Our findings demonstrate that, in humans, a miRNA loading complex (miRLC) is formed by Ago2 and Dicer prior to their encounter with pre miRNA. We suggest that the miRLC couples the processing of the pre-miRNA substrate to the unwinding of the product and that after loading of the mature miRNA to Ago2, the miRLC disassembles and the miRNP is released. PMID- 16357217 TI - Spliceosome disassembly catalyzed by Prp43 and its associated components Ntr1 and Ntr2. AB - Two novel yeast splicing factors required for spliceosome disassembly have been identified. Ntr1 and Ntr2 (NineTeen complex-Related proteins) were identified for their weak association with components of the Prp19-associated complex. Unlike other Prp19-associated components, these two proteins were primarily associated with the intron-containing spliceosome during the splicing reaction. Extracts depleted of Ntr1 or Ntr2 exhibited full splicing activity, but accumulated large amounts of lariat-intron in the spliceosome after splicing, indicating that the normal function of the Prp19-associated complex in spliceosome activation was not affected, but spliceosome disassembly was hindered. Immunoprecipitation analysis revealed that Ntr1 and Ntr2 formed a stable complex with DExD/H-box RNA helicase Prp43 in the splicing extract. Ntr1 interacted with Prp43 through the N-terminal G-patch domain, with Ntr2 through a middle region, and with itself through the carboxyl half of the protein. The affinity-purified Ntr1-Ntr2-Prp43 complex could catalyze disassembly of the spliceosome in an ATP-dependent manner, separating U2, U5, U6, NTC (NineTeen Complex), and lariat-intron. This is the first demonstration of physical disassembly of the spliceosome, catalyzed by a complex containing a DExD/H-box RNA helicase and two accessory factors, which might function in targeting the helicase to the correct substrate. PMID- 16357218 TI - The RNA polymerase II subunit Rpb4p mediates decay of a specific class of mRNAs. AB - It is commonly appreciated that the mRNA level is determined by the balance between its synthetic and decay kinetics. Yet, little is known about coordination between these distinct processes. A major pathway of the eukaryotic mRNA decay initiates with shortening of the mRNA poly(A) tail (deadenylation), followed by removal of the mRNA 5' cap structure and its subsequent exonucleolytic degradation. Here we report that a subunit of RNA polymerase II, Rpb4p, is required for the decay of a class of mRNAs whose products are involved in protein synthesis. Cells lacking RPB4 are defective in the deadenylation and post deadenylation steps of representatives of this class of mRNAs. Moreover, Rpb4p interacts with both the mRNP and with subunits of the mRNA decay complex Pat1/Lsm1-7 that enhances decapping. Consistently, a portion of Rpb4p is localized in P bodies, where mRNA decapping and degradation is executed, and mutations in RPB4 increase the number of P bodies per cell. We propose that Rpb4p has a dual function in mRNA decay. It promotes or enhances the deadenylation process of specific mRNAs and recruits Pat1/Lsm1-7 to these mRNAs, thus stimulating their decapping and further decay. In this way, Rpb4p might link the activity of the basal transcription apparatus with that of the mRNA decay machinery. PMID- 16357219 TI - The core centromere and Sgo1 establish a 50-kb cohesin-protected domain around centromeres during meiosis I. AB - The stepwise loss of cohesins, the complexes that hold sister chromatids together, is required for faithful meiotic chromosome segregation. Cohesins are removed from chromosome arms during meiosis I but are maintained around centromeres until meiosis II. Here we show that Sgo1, a protein required for protecting centromeric cohesins from removal during meiosis I, localizes to cohesin-associated regions (CARs) at the centromere and the 50-kb region surrounding it. Establishment of this Sgo1-binding domain requires the 120-base pair (bp) core centromere, the kinetochore component Bub1, and the meiosis specific factor Spo13. Interestingly, cohesins and the kinetochore proteins Iml3 and Chl4 are necessary for Sgo1 to associate with pericentric regions but less so for Sgo1 to associate with the core centromeric regions. Finally, we show that the 50-kb Sgo1-binding domain is the chromosomal region where cohesins are protected from removal during meiosis I. Our results identify the portions of chromosomes where cohesins are protected from removal during meiosis I and show that kinetochore components and cohesins themselves are required to establish this cohesin protective domain. PMID- 16357221 TI - Replisome instability, fork collapse, and gross chromosomal rearrangements arise synergistically from Mec1 kinase and RecQ helicase mutations. AB - The yeast checkpoint kinases Mec1 and Rad53 are required for genomic stability in the presence of replicative stress. When replication forks stall, the stable maintenance of replisome components requires the ATR kinase Mec1/Ddc2 and the RecQ helicase Sgs1. It was unclear whether either Mec1 or Sgs1 action requires the checkpoint effector kinase, Rad53. By combining sgs1Delta with checkpoint deficient alleles, we can now distinguish the role of Mec1 at stalled forks from that of Rad53. We show that the S-phase-specific mec1-100 allele, like the sgs1Delta mutation, partially destabilizes DNA polymerases at stalled forks, yet combining the mec1-100 and sgs1Delta mutations leads to complete disassociation of the replisome, loss of RPA, irreversible termination of nucleotide incorporation, and compromised recovery from hydroxyurea (HU) arrest. These events coincide with a dramatic increase in both spontaneous and HU-induced chromosomal rearrangements. Importantly, in sgs1Delta cells, RPA levels at stalled forks do not change, although Ddc2 recruitment is compromised, explaining the partial Sgs1 and Mec1 interdependence. Loss of Rad53 kinase, on the other hand, does not affect the levels of DNA polymerases at arrested forks, but leads to MCM protein dissociation. Finally, confirming its unique role during replicative stress, Mec1, and not Tel1, is shown to modify fork-associated histone H2A. PMID- 16357220 TI - The Rad50S allele promotes ATM-dependent DNA damage responses and suppresses ATM deficiency: implications for the Mre11 complex as a DNA damage sensor. AB - Genetic and cytologic data from Saccharomyces cerevisiae and mammals implicate the Mre11 complex, consisting of Mre11, Rad50, and Nbs1, as a sensor of DNA damage, and indicate that the complex influences the activity of ataxia telangiectasia mutated (ATM) in the DNA damage response. Rad50(S/S) mice exhibit precipitous apoptotic attrition of hematopoietic cells. We generated ATM- and Chk2-deficient Rad50(S/S) mice and found that Rad50(S/S) cellular attrition was strongly ATM and Chk2 dependent. The hypomorphic Mre11(ATLD1) and Nbs1(Delta)(B) alleles conferred similar rescue of Rad50(S/S)-dependent hematopoietic failure. These data indicate that the Mre11 complex activates an ATM-Chk2-dependent apoptotic pathway. We find that apoptosis and cell cycle checkpoint activation are parallel outcomes of the Mre11 complex-ATM pathway. Conversely, the Rad50(S) mutation mitigated several phenotypic features of ATM deficiency. We propose that the Rad50(S) allele is hypermorphic for DNA damage signaling, and that the resulting constitutive low-level activation of the DNA damage response accounts for the partial suppression of ATM deficiency in Rad50(S/S) Atm(-/-) mice. PMID- 16357222 TI - Inactivation of S6 ribosomal protein gene in T lymphocytes activates a p53 dependent checkpoint response. AB - Ribosome biogenesis has been associated with regulation of cell growth and cell division, but the molecular mechanisms that integrate the effect of ribosome biogenesis on these processes in mammalian cells remain unknown. To study the effect of impaired ribosome functions in vivo, we conditionally deleted one or two alleles of the 40S ribosomal protein S6 gene in T cells in the mouse. While complete deletion of S6 abrogated T-cell development, hemizygous expression did not have any effect on T-cell maturation in the thymus, but inhibited the accumulation of T cells in the spleen and lymph nodes, as a result of their decreased survival in the peripheral lymphoid organs. Additionally, TCR-mediated stimulation of S6-heterozygous T cells induced a normal increase in their size, but cell cycle progression was impaired. Genetic inactivation of p53 tumor suppressor rescued development of S6-homozygous null thymocytes and proliferative defect of S6-heterozygous T cells. These results demonstrate the existence of a p53-dependent checkpoint mechanism that senses changes in the fidelity of the translational machinery to prevent aberrant cell division or eliminate defective T cells in vivo. Failure to activate this checkpoint response could potentially lead to a development of pathological processes such as tumors and autoimmune diseases. PMID- 16357223 TI - Cell population heterogeneity during growth of Bacillus subtilis. AB - We have discovered that cells of Bacillus subtilis at the mid-exponential phase of growth are a mixed population of two strikingly different cell types. One type is single swimming cells (or cell doublets) in which the transcription factor for motility, sigma(D), is active (sigma(D) ON). The other type is long chains of sessile cells in which sigma(D) is inactive (sigma(D) OFF). The population is strongly biased toward sigma(D)-ON cells by the action of a novel regulatory protein called SwrA. SwrA stimulates the transcription of a large operon (the flagellum/chemotaxis operon), which includes the genes for sigma(D) and an activator of sigma(D)-directed gene expression, SwrB. Cell population heterogeneity could enable B. subtilis to exploit its present location through the production of sessile cells as well as to explore new environmental niches through the generation of nomadic cells. PMID- 16357224 TI - Scaling of connectivity in marine populations. AB - Defining the scale of connectivity, or exchange, among marine populations and determining the factors driving this exchange are pivotal to our understanding of the population dynamics, genetic structure, and biogeography of many coastal species. Using a high-resolution biophysical model for the Caribbean region, we report that typical larval dispersal distances of ecologically relevant magnitudes are on the scale of only 10 to 100 kilometers for a variety of reef fish species. We also show the importance of the early onset of active larval movement mediating the dispersal potential. In addition to self-recruitment, larval import from outside the local area is required to sustain most populations, although these population subsidies are very limited in particular systems. The results reveal distinct regions of population isolation based on larval dispersal that also correspond to genetic and morphological clines observed across a range of marine organisms. PMID- 16357225 TI - A stretchable form of single-crystal silicon for high-performance electronics on rubber substrates. AB - We have produced a stretchable form of silicon that consists of submicrometer single-crystal elements structured into shapes with microscale, periodic, wavelike geometries. When supported by an elastomeric substrate, this "wavy" silicon can be reversibly stretched and compressed to large levels of strain without damaging the silicon. The amplitudes and periods of the waves change to accommodate these deformations, thereby avoiding substantial strains in the silicon itself. Dielectrics, patterns of dopants, electrodes, and other elements directly integrated with the silicon yield fully formed, high-performance "wavy" metal oxide semiconductor field-effect transistors, p-n diodes, and other devices for electronic circuits that can be stretched or compressed to similarly large levels of strain. PMID- 16357226 TI - Femtosecond multidimensional imaging of a molecular dissociation. AB - The coupled electronic and vibrational motions governing chemical processes are best viewed from the molecule's point of view-the molecular frame. Measurements made in the laboratory frame often conceal information because of the random orientations the molecule can take. We used a combination of time-resolved photoelectron spectroscopy, multidimensional coincidence imaging spectroscopy, and ab initio computation to trace a complete reactant-to-product pathway-the photodissociation of the nitric oxide dimer-from the molecule's point of view, on the femtosecond time scale. This method revealed an elusive photochemical process involving intermediate electronic configurations. PMID- 16357227 TI - The snoRNA HBII-52 regulates alternative splicing of the serotonin receptor 2C. AB - The Prader-Willi syndrome is a congenital disease that is caused by the loss of paternal gene expression from a maternally imprinted region on chromosome 15. This region contains a small nucleolar RNA (snoRNA), HBII-52, that exhibits sequence complementarity to the alternatively spliced exon Vb of the serotonin receptor 5-HT(2C)R. We found that HBII-52 regulates alternative splicing of 5 HT(2C)R by binding to a silencing element in exon Vb. Prader-Willi syndrome patients do not express HBII-52. They have different 5-HT(2C)R messenger RNA (mRNA) isoforms than healthy individuals. Our results show that a snoRNA regulates the processing of an mRNA expressed from a gene located on a different chromosome, and the results indicate that a defect in pre-mRNA processing contributes to the Prader-Willi syndrome. PMID- 16357228 TI - Doing more for Kate. PMID- 16357229 TI - Stem cells. Korean university will investigate cloning paper. PMID- 16357230 TI - Space science. ESA hits the right note, and funding flows. PMID- 16357231 TI - Paleontology. How fast does your dinosaur grow? PMID- 16357232 TI - U.S. competitiveness. Summit lists ways--but not means--to strengthen science. PMID- 16357233 TI - After Katrina. Struggling New Orleans universities cut hundreds of faculty. PMID- 16357235 TI - Indian science. Booming computer sector seen as a mixed blessing. PMID- 16357234 TI - Genetics. Zebrafish researchers hook gene for human skin color. PMID- 16357236 TI - Scientific publishing. Echoing other cases, NEJM says Vioxx Safety data withheld. PMID- 16357237 TI - Spongiform diseases. After the crisis: more questions about prions. PMID- 16357238 TI - Spongiform diseases. Waiting for the final experiment. PMID- 16357239 TI - Conservation biology. Hawaii's coral trees feel the sting of foreign wasps. PMID- 16357240 TI - Environmental epigenomics meeting. Food, tobacco, and future generations. PMID- 16357241 TI - Environmental epigenomics meeting. Supplements restore gene function via methylation. PMID- 16357242 TI - Electronics. Organic LEDs look forward to a bright, white future. PMID- 16357243 TI - Asian scientists and the "glass ceiling". PMID- 16357244 TI - How to cut world hunger in half. PMID- 16357245 TI - Cognitive unbinding in sleep and anesthesia. PMID- 16357246 TI - Comment on "Characterization of excess electrons in water-cluster anions by quantum simulations". AB - The conclusion by Turi et al. (Reports, 5 August 2005, p. 914) that all experimental spectral and energetic data on water-cluster anions point toward surface-bound electrons is overstated. Comparison of experimental vertical detachment energies with their calculated values for (H2O)n- clusters with surface-bound and internalized electrons supports previous arguments that both types of clusters exist. PMID- 16357248 TI - Chemistry. Lateral hopping requires molecular rocking. PMID- 16357247 TI - Research funding. Science foundations: a novelty in Russian science. PMID- 16357249 TI - Microbiology. Chitin, cholera, and competence. PMID- 16357250 TI - Geophysics. Helium feels the heat in Earth's mantle. PMID- 16357251 TI - Ecology. Population evolution and island biogeography. PMID- 16357252 TI - Aphid protected from pathogen by endosymbiont. AB - Aphids are associated with several facultative bacterial endosymbionts that may influence their interactions with other organisms. We show here that one of the three most common facultative symbionts of pea aphid (Acyrthosiphon pisum), the bacterium Regiella insecticola, has a major effect on host resistance to a fungal pathogen. Experimental establishment of the bacterium in uninfected aphid clones led to higher survival after fungal attack. The bacteria also increased the aphid's inclusive fitness, because the presence of the symbiont reduced the probability of fungal sporulation on aphid cadavers, hence lowering the rate of transmission of the disease to nearby related aphids. PMID- 16357254 TI - Complete photo-induced breakup of the H2 molecule as a probe of molecular electron correlation. AB - Despite decades of progress in quantum mechanics, electron correlation effects are still only partially understood. Experiments in which both electrons are ejected from an oriented hydrogen molecule by absorption of a single photon have recently demonstrated a puzzling phenomenon: The ejection pattern of the electrons depends sensitively on the bond distance between the two nuclei as they vibrate in their ground state. Here, we report a complete numerical solution of the Schrodinger equation for the double photoionization of H2. The results suggest that the distribution of photoelectrons emitted from aligned molecules reflects electron correlation effects that are purely molecular in origin. PMID- 16357253 TI - SLC24A5, a putative cation exchanger, affects pigmentation in zebrafish and humans. AB - Lighter variations of pigmentation in humans are associated with diminished number, size, and density of melanosomes, the pigmented organelles of melanocytes. Here we show that zebrafish golden mutants share these melanosomal changes and that golden encodes a putative cation exchanger slc24a5 (nckx5) that localizes to an intracellular membrane, likely the melanosome or its precursor. The human ortholog is highly similar in sequence and functional in zebrafish. The evolutionarily conserved ancestral allele of a human coding polymorphism predominates in African and East Asian populations. In contrast, the variant allele is nearly fixed in European populations, is associated with a substantial reduction in regional heterozygosity, and correlates with lighter skin pigmentation in admixed populations, suggesting a key role for the SLC24A5 gene in human pigmentation. PMID- 16357255 TI - Multistep synthesis of a radiolabeled imaging probe using integrated microfluidics. AB - Microreactor technology has shown potential for optimizing synthetic efficiency, particularly in preparing sensitive compounds. We achieved the synthesis of an [(18)F]fluoride-radiolabeled molecular imaging probe, 2-deoxy-2-[18F]fluoro-D glucose ([18F]FDG), in an integrated microfluidic device. Five sequential processes-[18F]fluoride concentration, water evaporation, radiofluorination, solvent exchange, and hydrolytic deprotection-proceeded with high radio-chemical yield and purity and with shorter synthesis time relative to conventional automated synthesis. Multiple doses of [18F]FDG for positron emission tomography imaging studies in mice were prepared. These results, which constitute a proof of principle for automated multistep syntheses at the nanogram to microgram scale, could be generalized to a range of radiolabeled substrates. PMID- 16357256 TI - Direct experimental evidence of a growing length scale accompanying the glass transition. AB - Understanding glass formation is a challenge, because the existence of a true glass state, distinct from liquid and solid, remains elusive: Glasses are liquids that have become too viscous to flow. An old idea, as yet unproven experimentally, is that the dynamics becomes sluggish as the glass transition approaches, because increasingly larger regions of the material have to move simultaneously to allow flow. We introduce new multipoint dynamical susceptibilities to estimate quantitatively the size of these regions and provide direct experimental evidence that the glass formation of molecular liquids and colloidal suspensions is accompanied by growing dynamic correlation length scales. PMID- 16357257 TI - Developmental plasticity in the life history of a prosauropod dinosaur. AB - Long-bone histology indicates that the most common early dinosaur, the prosauropod Plateosaurus engelhardti from the Upper Triassic of Central Europe, had variable life histories. Although Plateosaurus grew at the fast rates typical for dinosaurs, as indicated by fibrolamellar bone, qualitative (growth stop) and quantitative (growth-mark counts) features of its histology are poorly correlated with body size. Individual life histories of P. engelhardti were influenced by environmental factors, as in modern ectothermic reptiles, but not in mammals, birds, or other dinosaurs. PMID- 16357258 TI - Drought, snails, and large-scale die-off of southern U.S. salt marshes. AB - Salt marshes in the southeastern United States have recently experienced massive die-off, one of many examples of widespread degradation in marine and coastal ecosystems. Although intense drought is thought to be the primary cause of this die-off, we found snail grazing to be a major contributing factor. Survey of marsh die-off areas in three states revealed high-density fronts of snails on die off edges at 11 of 12 sites. Exclusion experiments demonstrated that snails actively converted marshes to exposed mudflats. Salt addition and comparative field studies suggest that drought-induced stress and grazers acted synergistically and to varying degrees to cause initial plant death. After these disturbances, snail fronts formed on die-off edges and subsequently propagated through healthy marsh, leading to cascading vegetation loss. These results, combined with model analyses, reveal strong interactions between increasing climatic stress and grazer pressure, both potentially related to human environmental impacts, which amplify the likelihood and intensity of runaway collapse in these coastal systems. PMID- 16357259 TI - Island biogeography of populations: an introduced species transforms survival patterns. AB - Population phenomena, which provide much of the underlying basis for the theoretical structure of island biogeography, have received little direct study. We determined a key population trait-survival-in the Bahamian lizard Anolis sagrei on islands with an experimentally introduced predatory lizard and on neighboring unmanipulated islands. On unmanipulated islands, survival declined with several variables, most notably vegetation height: The island with the shortest vegetation had nearly the highest survival recorded for any lizard. On islands with the introduced predator, which forages mostly on the ground, A. sagrei shifted to taller vegetation; unlike on unmanipulated islands, its survival was very low on islands with the shortest vegetation but was higher on the others. Thus, species introduction radically changed a resident species' relation of survival to a key island-biogeographical variable. PMID- 16357260 TI - Long-term modulation of electrical synapses in the mammalian thalamus. AB - Electrical synapses are common between inhibitory neurons in the mammalian thalamus and neocortex. Synaptic modulation, which allows flexibility of communication between neurons, has been studied extensively at chemical synapses, but modulation of electrical synapses in the mammalian brain has barely been examined. We found that the activation of metabotropic glutamate receptors, via endogenous neurotransmitter or by agonist, causes long-term reduction of electrical synapse strength between the inhibitory neurons of the rat thalamic reticular nucleus. PMID- 16357261 TI - Ubiquitin-binding domains in Y-family polymerases regulate translesion synthesis. AB - Translesion synthesis (TLS) is the major pathway by which mammalian cells replicate across DNA lesions. Upon DNA damage, ubiquitination of proliferating cell nuclear antigen (PCNA) induces bypass of the lesion by directing the replication machinery into the TLS pathway. Yet, how this modification is recognized and interpreted in the cell remains unclear. Here we describe the identification of two ubiquitin (Ub)-binding domains (UBM and UBZ), which are evolutionarily conserved in all Y-family TLS polymerases (pols). These domains are required for binding of poleta and poliota to ubiquitin, their accumulation in replication factories, and their interaction with monoubiquitinated PCNA. Moreover, the UBZ domain of poleta is essential to efficiently restore a normal response to ultraviolet irradiation in xeroderma pigmentosum variant (XP-V) fibroblasts. Our results indicate that Ub-binding domains of Y-family polymerases play crucial regulatory roles in TLS. PMID- 16357262 TI - Chitin induces natural competence in Vibrio cholerae. AB - The mosaic-structured Vibrio cholerae genome points to the importance of horizontal gene transfer (HGT) in the evolution of this human pathogen. We showed that V. cholerae can acquire new genetic material by natural transformation during growth on chitin, a biopolymer that is abundant in aquatic habitats (e.g., from crustacean exoskeletons), where it lives as an autochthonous microbe. Transformation competence was found to require a type IV pilus assembly complex, a putative DNA binding protein, and three convergent regulatory cascades, which are activated by chitin, increasing cell density, and nutrient limitation, a decline in growth rate, or stress. PMID- 16357263 TI - Invited review: Methods to determine body fat reserves in the dairy cow with special regard to ultrasonographic measurement of backfat thickness. AB - As the dairy cow uses body energy reserves in early lactation, body condition scoring has become an integral part of dairy herd management. Several methods based on visual and tactile evaluation have been developed. Problems caused by the subjectivity of these techniques have been reported. Alternative approaches to predict energy reserves or energy balance in dairy cattle include metabolic profiling and measurement of live weight, heart girth, or skinfold thickness. A less common method to assess fat reserves in body tissues is measuring backfat thickness (BFT) by using ultrasound. An ultrasound technique has been established to predict carcass quality in beef cattle. A new aspect is the application of ultrasound as a monitoring tool in dairy herd management where another location has to be evaluated. This technique has been validated by relating BFT to total body fat (TBF) content and carcass BFT. Backfat thickness also has been related to other methods of body condition scoring. Target values for the development of BFT throughout lactation are available. The relationship between BFT and TBF content is highly significant although biased by multiple factors. A change in BFT of 1 mm equates to approximately 5 kg of TBF content. Measuring BFT by ultrasound is of added value compared with other body condition scoring systems because it is objective and precise. Changes in body condition can be detected and evaluated properly. PMID- 16357264 TI - Effect of trisodium citrate concentration and cooking time on the physicochemical properties of pasteurized process cheese. AB - The effects of the concentration of trisodium citrate (TSC) emulsifying salt (0.25 to 2.75%) and holding time (0 to 20 min) on the textural, rheological, and microstructural properties of pasteurized process Cheddar cheese were studied using a central composite rotatable design. The loss tangent parameter (from small amplitude oscillatory rheology), extent of flow (derived from the University of Wisconsin Meltprofiler), and melt area (from the Schreiber test) all indicated that the meltability of process cheese decreased with increased concentration of TSC and that holding time led to a slight reduction in meltability. Hardness increased as the concentration of TSC increased. Fluorescence micrographs indicated that the size of fat droplets decreased with an increase in the concentration of TSC and with longer holding times. Acid-base titration curves indicated that the buffering peak at pH 4.8, which is due to residual colloidal calcium phosphate, decreased as the concentration of TSC increased. The soluble phosphate content increased as concentration of TSC increased. However, the insoluble Ca decreased with increasing concentration of TSC. The results of this study suggest that TSC chelated Ca from colloidal calcium phosphate and dispersed casein; the citrate-Ca complex remained trapped within the process cheese matrix. Increasing the concentration of TSC helped to improve fat emulsification and casein dispersion during cooking, both of which probably helped to reinforce the structure of process cheese. PMID- 16357265 TI - Use of the Foodtexture Puff Device to monitor milk coagulation. AB - The further automation of cheese-making on an industrial level requires the development of sensor devices to monitor the gelation process and especially the firming phase. In this paper, the Foodtexture Puff Device (FPD) is tested for its ability to monitor the gelation process by comparing it with classical rheometry (G' and G'') in a series of coagulations at different initial milk pH (6.01 to 6.61). The FPD measures the deformation of the surface of the milk during coagulation after applying an air puff directed on this surface. The maximal and minimal deformation values and the deformation range were calculated. A nonlinear model of the registered characteristics with the time point from adding rennet until the end of the gelation process was fitted on the FPD data and also on the classic rheology parameters. It was concluded that the FPD monitored the coagulation process in the same way as the rheology. Moreover, the start point of the coagulation process as well as the strength of the coagulum could be estimated nondestructively. Therefore, the presented technology together with the nonlinear model may be a basis for the development of an industrial monitoring device. PMID- 16357266 TI - Proteolysis and microstructure of Piacentinu Ennese cheese made using different farm technologies. AB - The aim of this study was to provide the biochemical and structural characterization of Piacentinu Ennese cheese and to evaluate the impact of different farm technologies on cheese proteolysis and microstructure. Fifteen cheeses were manufactured according to traditional technology, i.e., from raw milk and farmhouse rennet in the absence of starter culture. Pasteurized milk, commercial rennet, and starter were used for production of 20 nontraditional cheeses. Proteolysis in Piacentinu Ennese cheese was monitored during a 2- to 10 mo ripening time. Low rates of overall proteolysis were observed in cheese, as percentages of total N soluble at pH 4.6 and in 12% trichloroacetic acid were about 11.40 and 8.10%, respectively, after 10 mo of age. Patterns of primary proteolysis by urea-PAGE showed that alpha(s)-caseins were degraded to a larger extent than were beta-caseins, although a considerable amount of both caseins was still intact after 10 mo. Reversed phase-HPLC analysis of the cheese peptide fractions showed a slow decrease in the levels of hydrophobic peptides coupled to increasing levels of hydrophilic compounds as the cheese aged. The structural characteristics of Piacentinu Ennese cheese were evaluated by scanning electron microscopy after 2, 4, and 6 mo of age. The micrographs showed a sponge-like structural network with a well-distributed system of empty spaces, originally occupied by whey and fat. The microstructure changed during cheese ripening to become more compact with cavities of smaller size. Farm technology significantly affected cheese proteolysis and microstructure. Nontraditional cheeses had higher levels of pH 4.6-soluble N and showed a larger hydrolysis of alpha(s)-casein fractions by urea-PAGE analysis than did traditional cheeses. Large differences between cheese-types also concerned the patterns of secondary proteolysis. Nontraditional cheeses had higher levels of 12% trichloroacetic acid-soluble N and showed larger proportions of free amino acids and hydrophilic peptides in the HPLC profiles of the corresponding 70% ethanol-soluble N fraction than traditional cheeses. Nontraditional cheeses also had a more open structure with a coarser and less continuous appearance than did traditional cheeses. A large amount of variability in cheese proteolysis and structure within nontraditional treatment reflected farm-dependent changes in manufacturing conditions related to the use of various types of rennet and starter. PMID- 16357267 TI - Ice recrystallization inhibition in ice cream as affected by ice structuring proteins from winter wheat grass. AB - Ice recrystallization in quiescently frozen sucrose solutions that contained some of the ingredients commonly found in ice cream and in ice cream manufactured under commercial conditions, with or without ice structuring proteins (ISP) from cold-acclimated winter wheat grass extract (AWWE), was assessed by bright field microscopy. In sucrose solutions, critical differences in moisture content, viscosity, ionic strength, and other properties derived from the presence of other ingredients (skim milk powder, corn syrup solids, locust bean gum) caused a reduction in ice crystal growth. Significant ISP activity in retarding ice crystal growth was observed in all solutions (44% for the most complex mix) containing 0.13% total protein from AWWE. In heat-shocked ice cream, ice recrystallization rates were significantly reduced 40 and 46% with the addition of 0.0025 and 0.0037% total protein from AWWE. The ISP activity in ice cream was not hindered by its inclusion in mix prior to pasteurization. A synergistic effect between ISP and stabilizer was observed, as ISP activity was reduced in the absence of stabilizer in ice cream formulations. A remarkably smoother texture for ice creams containing ISP after heat-shock storage was evident by sensory evaluation. The efficiency of ISP from AWWE in controlling ice crystal growth in ice cream has been demonstrated. PMID- 16357268 TI - Fluorescent labeling study of plasminogen concentration and location in simulated bovine milk systems. AB - A fluorescent labeling method was developed to study plasminogen (PG) concentration and location in simulated bovine milk. Activity and stability of PG labeled with Alexa Fluor 594 (PG-594) were comparable to those of native PG. The fluorescent signal of PG-594 exhibited pH, temperature, and storage stability, and remained stable throughout typical sample treatments (stirring, heating, and ultracentrifugation). These characteristics indicate broad applicability of the fluorescent labeling technique for milk protease characterization. In an example application, PG-594 was added to simulated milk samples to study effects of heat and beta-lactoglobulin (beta-LG) on the distribution of PG. Before heating, about one-third of the PG-594 remained soluble in the whey fraction (supernatant) whereas the rest became associated with the casein micelle. Addition of beta-LG to the system slightly shifted PG-594 distribution toward the whey fraction. Heat induced PG-594 binding to micelles in whey-protein-free systems was evidenced by a decrease of PG-594 from 31 to 15% in the whey fraction accompanied by an increase of PG-594 from 69 to 85% in casein micelle fractions. When beta-LG was present during heating, more than 95% of PG-594 became associated with the micelle. A comparison with the distribution pattern of PG-derived activities revealed that heat-induced PG binding to micelles accompanies heat-induced PG inactivation in the micelle fraction. Incubation of the casein micelles with the reducing agent beta-mercaptoethanol revealed that disulfide bonds formed between PG and casein or between PG and casein-bound beta-LG are the mechanisms for heat induced PG binding to casein micelles. Western blotting and zymography results correlated well with fluorescent labeling studies and activity studies, respectively. Theoretically important findings are: 1) when heated, serum PG is capable of covalently binding to micellar casein or complexing with beta-LG in whey and then coadhering to micelles, and 2) PG that associated with micellar casein through lysine binding sites before heating is capable of developing heat induced disulfide bonds with casein. The overall results are PG covalently binding to micelles and inactivation thereafter. Our results suggest that, instead of thermal denaturation through irreversible unfolding, covalent bond formation between PG and other milk proteins is the mechanism of PG inhibition during thermal processing. PMID- 16357270 TI - Indirect chiral HPLC determination and fluorimetric detection of D-amino acids in milk and oyster samples. AB - The indirect chiral method, based on the o-phthaldialdehyde reaction using the chiral N-acetyl-L-cysteine reagent, has been optimized to determine D-amino acids in milk and oyster samples. Both the derivatization reaction and the HPLC separation have been improved, and simple sample treatments were proposed. The milk sample preparation involved centrifugation and filtration through polytetrafluoroethylene filters for determination of free D-AA. Two methods, nonenzymatic and enzymatic, were applied to determination of free and total D-AA, respectively, in oyster samples. The D-AA contents were in the range of 0.14 to 4.32 mg/L for milk samples and 0.80 to 15.9 mg/g for oyster samples, with relative standard deviations lower than 10% (n = 4), except for D-Tyr. In general, mean recoveries were greater than 90% with relative standard deviations lower than 9% (n = 4) at concentration levels from 0.055 to 0.426 mg/L for milk samples and 0.348 to 1.10 mg/ g for oyster samples. PMID- 16357269 TI - Properties of whey protein isolates extruded under acidic and alkaline conditions. AB - Whey proteins have wide acceptance and use in many products due to their beneficial nutritional properties. To further increase the amount of whey protein isolates (WPI) that may be added to products such as extruded snacks and meats, texturization of WPI is necessary. Texturization changes the folding of globular proteins to improve interaction with other ingredients and create new functional ingredients. In this study, WPI pastes (60% solids) were extruded in a twin-screw extruder at 100 degrees C with 4 pH-adjusted water streams: acidic (pH 2.0 +/- 0.2) and alkaline (pH 12.4 +/- 0.4) streams from 2 N HCl and 2 N NaOH, respectively, and acidic (pH 2.5 +/- 0.2) and alkaline (pH 11.5 +/- 0.4) electrolyzed water streams; these were compared with WPI extruded with deionized water. The effects of water acidity on WPI solubility at pH 7, color, microstructure, Rapid Visco Analyzer pasting properties, and physical structure were determined. Alkaline conditions increased insolubility caused yellowing and increased pasting properties significantly. Acidic conditions increased solubility and decreased WPI pasting properties. Subtle structural changes occurred under acidic conditions, but were more pronounced under alkaline conditions. Overall, alkaline conditions increased denaturation in the extruded WPI resulting in stringy texturized WPI products, which could be used in meat applications. PMID- 16357271 TI - Technical note: Improved extraction method with hexane for gas chromatographic analysis of conjugated linoleic acids. AB - Extraction properties of different solvents (chloroform/methanol, hexane/isopropanol, and hexane) were studied for the gas chromatographic analysis of conjugated linoleic acids (CLA) from probiotic bacteria grown in de Man, Rogosa, and Sharpe medium. As compared with chloroform/methanol and hexane/isopropanol, hexane showed comparable extraction efficiency for CLA from unspent de Man, Rogosa, and Sharpe medium, but showed minimal extraction of oleic acid originated from the emulsifier in broth. The extraction efficiency of CLA by hexane was influenced by the broth pH, showing the optimal pH of 7.0. Repeated extraction with hexane increased the yield. Extraction with hexane showed excellent recovery of spiked CLA from the spent broth with up to 97.2% (standard deviation of 1.74%). This represents the highest recovery of CLA from culture broth ever reported. The sample size was also successfully reduced to 0.5 mL to analyze CLA from the broth without impairment of analytical data. This smaller sample size in the 1.5-mL microcentrifuge tube using a small bench-top centrifuge reduced analytical time significantly. PMID- 16357272 TI - Improvement of texture and structure of reduced-fat Cheddar cheese by exopolysaccharide-producing lactococci. AB - The objective of this study was to evaluate the effect of capsular and ropy exopolysaccharide (EPS)-producing strains of Lactococcus lactis ssp. cremoris on textural and microstructural attributes during ripening of 50%-reduced-fat Cheddar cheese. Cheeses were manufactured with added capsule- or ropy-forming strains individually or in combination. For comparison, reduced-fat cheese with or without lecithin added at 0.2% (wt/vol) to cheese milk and full-fat cheeses were made using EPS-nonproducing starter, and all cheeses were ripened at 7 degrees C for 6 mo. Exopolysaccharide-producing strains increased cheese moisture retention by 3.6 to 4.8% and cheese yield by 0.28 to 1.19 kg/100 kg compared with control cheese, whereas lecithin-containing cheese retained 1.4% higher moisture and had 0.37 kg/100 kg higher yield over the control cheese. Texture profile analyses for 0-d-old cheeses revealed that cheeses with EPS-producing strains had less firm, springy, and cohesive texture but were more brittle than control cheeses. However, these effects became less pronounced after 6 mo of ripening. Using transmission electron microscopy, fresh and aged cheeses with added EPS producing strains showed a less compact protein matrix through which larger whey pockets were dispersed compared with control cheese. The numerical analysis of transmission electron microscopy images showed that the area in the cheese matrix occupied by protein was smaller in cheeses with added EPS-producing strains than in control cheese. On the other hand, lecithin had little impact on both cheese texture and microstructure; after 6 mo, cheese containing lecithin showed a texture profile very close to that of control reduced-fat cheese. The protein occupied area in the cheese matrix did not appear to be significantly affected by lecithin addition. Exopolysaccharide-producing strains could contribute to the modification of cheese texture and microstructure and thus modify the functional properties of reduced-fat Cheddar cheese. PMID- 16357273 TI - Effects of feeding propionibacteria to dairy cows on milk yield, milk components, and reproduction. AB - Two weeks before parturition, 38 Holstein primiparous and multiparous cows were assigned to 1 of 3 treatment groups: control animals (n = 13) received regular total mixed rations (TMR), the low-dose group (n = 14) received the control TMR plus 6 x 10(10) cfu/cow of Propionibacterium strain P169 (P169), and the high dose group (n = 11) received the control TMR plus 6 x 10(11) cfu/cow of P169 from -2 to 30 wk postpartum. Weekly milk samples were analyzed for percentage of milk fat, protein, lactose, and SNF, milk urea nitrogen, and somatic cell counts. Daily milk production expressed as 4% fat-corrected milk was affected by treatment and week x parity. High-dose and low-dose P169-treated cows exhibited 7.1 and 8.5% increases above controls in daily 4% fat-corrected milk, respectively. Treatment x parity and week significantly influenced percentage of milk fat, lactose, and protein, whereas treatment x parity and treatment x week influenced SNF. Ruminal propionate levels were influenced by treatment such that high-dose P169 cows had greater molar percentage of propionate than did low-dose P169 and control cows. Change in body weight postpartum was influenced by week x parity and treatment x parity such that high-dose and low-dose P169 multiparous cows exhibited a more rapid recovery of wk-1 body weight than did control multiparous cows. There was no treatment, parity, or interaction on days to first postpartum ovulation or on estrous behavior at 45 and 90 d postpartum. We concluded that P169 might have potential as an effective direct-fed microorganism to increase milk production in dairy cows. PMID- 16357274 TI - Stocking density and feed barrier design affect the feeding and social behavior of dairy cattle. AB - The objectives of this study were to: 1) evaluate how stocking density at the feed bunk affects feeding and social behavior of dairy cows; and 2) determine if this effect is further influenced by the type of feed barrier used. Thirty-six lactating Holstein cows, allotted to 4 groups, were subjected to each of 4 stocking density treatments and 2 feed barrier treatments. Initially, 2 groups were assigned to a headlock barrier, and 2 groups to a post-and-rail barrier. Each group was then exposed to 4 stocking density treatments (0.81, 0.61, 0.41, and 0.21 m/cow, corresponding to 1.33, 1.00, 0.67, and 0.33 headlocks/cow), in 4 successive 10-d treatment periods. After these periods, the feed barriers were switched between groups and the 4 stocking density treatments were readministered. Time-lapse video was used to quantify feeding, standing, and aggressive behavior at the feed bunk. Daily feeding times were greater and duration of inactive standing in the feeding area was less when using a post-and rail compared with a headlock feed barrier. Feeding time decreased and inactive standing increased for both barrier designs as stocking density increased at the feed bunk. Cows were displaced more often from the feeding area when the stocking density was increased, and this effect was greater for cows using the post-and rail feed barrier. Cows ranked lower in the social hierarchy at the feed bunk were displaced more often when feeding at a post-and-rail barrier, particularly at high stocking densities. Therefore, we recommend avoiding overstocking at the feed bunk to increase feeding activity and reduce competition. Use of a barrier that provides some physical separation between adjacent cows, such as a headlock feed barrier, can be used to further reduce competition at the feed bunk. PMID- 16357275 TI - Short communication: Feed bunk utilization in dairy cows housed in pens with either two or three rows of free stalls. AB - The goal of this observational study was to determine whether there were significant differences in feed bunk utilization patterns of dairy cows in free stall pens with either 2 or 3 rows of stalls. Previously recorded 24-h videos of the high-yielding, mature cow group in 5 herds with 2-row pen designs and 5 herds with 3-row pen designs that were provided fresh TMR once a day after the morning milking were scan-sampled at 10-min intervals to record feed bunk utilization score (proportion of feed bunk spaces in the pen that were filled). Data were aligned according to peak feed bunk utilization score following fresh feed delivery (primary peaks), return from the afternoon milking (secondary peaks), and return from the night milking (tertiary peaks). Bunk utilization score was highest for primary peaks; scores were not significantly different between 2- and 3-row pens (0.65 vs. 0.71 +/- 0.1, respectively), and rates of decline in bunk utilization score were similar over the ensuing 90-min period between pen designs. Differences in bunk utilization between peak type and pen row type were observed; tertiary bunk utilization with 2-row pens was significantly lower than bunk utilization with 3-row pens and significantly lower than either primary or secondary bunk utilization in 2-row pens. The feed space allowance provided by a 2-row pen design potentially allows cows to demonstrate other important feeding behaviors, such as avoidance of conflicts and maintenance of greater inter-cow distances between neighbors while feeding. PMID- 16357276 TI - Effect of hoof pathologies on subjective assessments of dairy cow gait. AB - To explore how hoof pathologies affect dairy cattle gait, we studied cows with sole hemorrhages (n = 14), sole ulcers (n = 7), and those with no visible injuries (n = 17). Overall gait assessments, scored from video using a 1 to 5 numerical rating system (1 = sound, 5 = severely lame) and a continuous 100-unit visual analog scale, found cows having sole ulcers had poorer gait than healthy cows (mean +/- SEM: 4.0 +/- 0.13 vs. 3.1 +/- 0.08, and 59 +/- 3 vs. 46 +/- 2, respectively). Six gait attributes (back arch, head bob, tracking-up, joint flexion, asymmetric gait, and reluctance to bear weight) were also assessed using continuous 100-unit scales. Compared with healthy cows, those having sole ulcers walked with a more pronounced back arch (12 +/- 3 vs. 28 +/- 4), more jerky head movement (2 +/- 2 vs. 10 +/- 3), shortened strides (7 +/- 2 vs. 26 +/- 4), and more uneven weighting among the limbs (16 +/- 2 vs. 32 +/- 3). Of all measures, the numerical rating system most effectively discriminated healthy cows from those with sole ulcers (R2 = 0.73), classifying 92% of animals correctly. No differences were detected among cows with and without sole hemorrhages. Intra- and interobserver reliabilities were reasonable for all measures (R2 > or = 0.64) except joint flexion and asymmetric gait. In summary, subjective assessments of dairy cattle gait provide valid and reliable approaches to identifying cattle with sole ulcers. PMID- 16357277 TI - In vitro effects of nonesterified fatty acids on bovine neutrophils oxidative burst and viability. AB - An in vitro study was conducted to examine the influence of nonesterified fatty acids (NEFA) on bovine polymorphonuclear leukocytes (PMN). Eight healthy, midlactating Holstein cows were used as blood donors. Blood PMN were isolated and incubated with a mixture of NEFA, reflecting composition of bovine plasma NEFA at concentrations that were intended to mimic those found in blood of cows undergoing high, moderate, or low lipomobilization intensity (2, 1, 0.5, 0.25, 0.125, and 0.0625 mM). Control samples were incubated in absence of NEFA. Phagocytosis and oxidative burst activities were assessed by a 2-color flow cytometric method, which was based on oxidation of intracellular dihydrorhodamine 123 to green fluorescent rhodamine 123. Oxidative burst products were generated by incubating PMN with Staphylococcus aureus labeled with propidium iodide. A flow cytometric technique was used to detect PMN viability, necrosis, and apoptosis using fluorescein isothiocyanate-labeled annexin-V and propidium iodide. Phagocytic activity was not affected by NEFA. The highest concentration of NEFA (2 mM) was associated with a dramatic increase of phagocytosis-associated oxidative burst activities with a reduction in cell viability (48.0 vs. 97.5% in control samples) and with a marked increase of necrosis (49.4 vs. 0.5% in control samples). Conversely, the mixture of NEFA did not affect the occurrence of apoptosis. Enhancement of the oxidative burst associated with the highest concentration of NEFA might explain the reduced viability and higher percentage of necrosis observed under the same conditions. This study demonstrated a substantial resistance of bovine PMN to an overload of fatty acids. However, observation that the highest concentration of NEFA regulated some PMN functions encourages the possibility of in vivo studies to assess the relationships between intensity of lipomobilization, plasma NEFA, and bovine PMN functions. PMID- 16357278 TI - Slatted floors and solid floors: stress and strain on the bovine hoof capsule analyzed in finite element analysis. AB - An established finite element model of a bovine claw was used to compare mechanical stress levels in a loaded model claw on different types of flooring. The following situations were compared: a claw standing on a solid floor, a claw standing on the edge of a short tie stand, and claws standing on slatted floors with slats of 28 and 40 mm (wide) running parallel and perpendicular to the claw axis. Finite element analysis allowed visualization of stress peaks seen predominantly in the weight-bearing border of the dorsal abaxial wall and of the bulbar region and in the proximal axial wall. Maximum stress values of 13 MPa were found in the model claw loaded on the solid floor and values of 18 to 22 MPa were seen in the model claw loaded on the edge of the solid floor. On slatted floors, stresses increased in the situation in which the claw was not supported under the abaxial wall. Comparison between the other slatted floors showed little difference in amounts of mechanical stress. A clear distinction was detected between the solid floor with full claw contact and the slatted floors. From the point of view of the mechanical stress seen in finite element analysis, a large contact area between claw and floor, as seen in the solid surface floor, is preferable. When use of slatted floors is unavoidable, direction of the slats should run perpendicular to the direction of the walkway to prevent even more mechanical impact in certain footing situations. PMID- 16357279 TI - Technical note: Improved method for rapid DNA extraction of mastitis pathogens directly from milk. AB - Efficient control against bovine mastitis requires sensitive, rapid, and specific tests to detect and identify the main bacteria that cause heavy losses to the dairy industry. Molecular detection of pathogenic microorganisms is based on DNA amplification of the target pathogen. Therefore, efficient extraction of DNA from pathogenic bacteria is a major step. In this study, we aimed to develop a specific, sensitive, and rapid method to extract DNA directly from the main gram positive bacteria known to cause bovine mastitis (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis) found in milk samples. The DNA extraction method is based on the lysing and nuclease-inactivating properties of the chaotropic agent, guanidinium thiocyanate, together with the nucleic acid-binding properties of the silica particles. An efficient protocol consisting of 6 basic steps (3 of which were done twice) was developed and applied directly to milk samples. Absence of PCR inhibitors and DNA quality were evaluated by PCR amplification of the species specific DNA sequences of the target bacteria. The level of sensitivity achieved in our experiments is applicable to milk sample analysis without sample enrichment. PMID- 16357280 TI - Short communication: The nature of heptadecenoic acid in ruminant fats. AB - Heptadecenoic acid (17:1) is a minor constituent of ruminant fats and its isomeric definition remains undefined in most reports on ruminant milk and intramuscular fat. Samples of milk and intramuscular fat of bovine, ovine, and caprine origin were analyzed by gas chromatography (GC) using 3 capillary columns with and without addition of 17:1 cis-10. Additionally, cis isomers of ovine milk fat samples were isolated as methyl esters by preparative thin-layer chromatography and analyzed by GC. The structural analysis of 17:1 present in samples was achieved by chemical ionization tandem mass spectrometry techniques. The isomer 17:1 cis-9 is the overwhelming heptadecenoic isomer in ruminant milk and intramuscular fat; 17:1 cis-10 is virtually absent. Moreover, current GC methods were able to resolve cis-9 from cis-10 and cis-8 isomers, so reports on 17:1 contents in ruminant fat should define its isomeric composition. PMID- 16357281 TI - Dietary forage and nonfiber carbohydrate contents influence B-vitamin intake, duodenal flow, and apparent ruminal synthesis in lactating dairy cows. AB - The objective of this experiment was to quantify intakes, duodenal flows, and ruminal apparent synthesis (AS) of B-vitamins in lactating dairy cows fed diets varying in forage and nonfiber carbohydrate (NFC) contents. Eight (4 primiparous and 4 multiparous) ruminally and duodenally cannulated Holstein cows were assigned to 4 dietary treatments in a replicated 21-d period, 4 x 4 Latin square design with a 2 x 2 factorial treatment arrangement. Diets, fed as TMR, contained (DM basis) 2 levels of forage (35 and 60%) and 2 levels of NFC (30 and 40%). The forage portion of the diets contained 50% corn silage, 33% alfalfa hay, and 17% grass hay. Soybean hulls and beet pulp (2:1) and corn meal and ground barley (2:1) were included to achieve desired NFC concentrations. No supplemental B vitamins were fed. B-vitamin AS was calculated as the amount of a specific B vitamin flowing to the duodenum minus its daily orts-corrected intake. Dry matter and organic matter intakes were higher for cows fed the 35% forage diets and the 40% NFC diets. Increasing dietary forage content decreased ruminal AS of pyridoxine, folic acid, and B12. Increasing dietary NFC content increased ruminal AS of nicotinic acid, nicotinamide, niacin, pyridoxal, B6, and folic acid but decreased AS of B12. Across diets, amounts of B-vitamins synthesized were highest for niacin, followed by riboflavin, B12, thiamin, B6, and folic acid. Biotin AS values were negative for all diets, suggesting either no ruminal synthesis or that destruction by ruminal microflora was greater than synthesis. B-vitamin intake, duodenal flow, and ruminal synthesis are influenced by dietary forage and NFC contents. PMID- 16357282 TI - Lipid metabolite profiles and milk production for Holstein and Jersey cows fed rumen-protected choline during the periparturient period. AB - Choline is important for assembly of very low density lipoproteins to export triglyceride from liver; however, studies to assess the effect of rumen-protected choline (RPC) supplementation on blood lipid metabolites in periparturient dairy cows have not been conducted. Thirty-two multiparous Holstein and 10 multiparous Jersey cows were randomly assigned to control or RPC treatments. A close-up diet was fed from approximately 3 wk before parturition through parturition, followed by a lactation diet from parturition through 49 d postpartum. For RPC, diets were top-dressed once daily with 60 g of a RPC product (25% choline as choline chloride) from 21 d before expected parturition through 21 d postpartum. Treatment did not affect dry matter intake either prepartum (12.0 vs. 12.1 kg/d for RPC and control, respectively) or during the first 3 wk postpartum (14.8 vs. 15.7 kg/d, respectively). Daily yields of 3.5% fat-corrected milk (39.4 vs. 37.4 kg/d), fat (1.46 vs. 1.38 kg/d), and protein (1.09 vs. 1.05 kg/d) did not differ statistically by treatment (RPC vs. control, respectively). Jersey cows in the control group had lower concentrations of nonesterified fatty acids and beta hydroxybutyrate in plasma during d 1 to 10 postpartum than did other breed and treatment combinations. Cows fed RPC tended to have greater serum triglycerides prepartum (17.0 vs. 14.7 mg/dL) and lower plasma phospholipid at parturition (65.2 vs. 78.1 mg/dL) than control cows. Treatment did not affect cholesterol and phospholipid at other time points, but concentrations followed patterns of dry matter intake pre- and postpartum. Cows were in moderate body condition score (mean = 3.3) at the start of the study and did not lose excessive condition by 3 wk postpartum (mean body condition score loss = 0.5); therefore, cows might not have been at great risk for hepatic lipid accumulation. Additionally, calculated Met balance was negative postpartum; supplemental RPC might not have spared enough Met to produce a physiological benefit. More research is needed to determine how choline affects prevention or alleviation of fatty liver syndrome and to confirm potential differences between Holstein and Jersey cows. PMID- 16357283 TI - Computer-controlled milk feeding of group-housed calves: the effect of milk allowance and weaning type. AB - Calves fed by computer-controlled milk feeders are often weaned gradually by reducing the size of the milk portions. However, reducing the number of milk portions instead may lower calves' occupation of the milk feeder and stimulate their concentrate intake, especially when they are offered a high milk allowance. Before weaning, but not during weaning, the calves on low milk allowance occupied the milk feeder more, consumed more concentrates, and had a lower daily gain. There was no interaction between milk allowance and weaning type. Weaning by reducing the number of milk portions resulted in more unrewarded visits to the milk feeder, but less time ingesting a similar amount of milk. The effect of milk allowance on milk feeder occupancy before weaning was not found when this allowance was halved during weaning. PMID- 16357284 TI - Effects of additional milk replacer feeding on calf health, growth, and selected blood metabolites in calves. AB - The objective of the experiment was to evaluate effects of increased milk replacer feeding on growth, intake, feed efficiency, and health parameters in stressed calves. Holstein bull calves (n = 120; approximately 3 to 8 d of age) were purchased from sale barns and dairy farms and housed in fiberglass hutches. In addition, wood shavings contaminated with coronavirus were mixed with clean shavings and added to each hutch before the start of the experiment. Calves were fed either a fixed amount (454 g/d) of a 20% crude protein (CP), 20% fat milk replacer to weaning at 28 d or a variable amount (454, 681, 908, and 454 g/d on d 0 to 7, 8 to 14, 15 to 31, and 32 to 41, respectively) of a milk replacer containing 28% CP and 17% fat without or with added dietary supplement containing bovine serum. Calves were also fed commercial calf starter and water ad libitum. Plasma IgG concentration in most calves on arrival at the facility was < 10 g/L. Intake, change in body weight, feed efficiency, morbidity and mortality, and selected plasma metabolites were determined. Body weight at 28 d, 56 d, daily body weight gain, intake of milk replacer, fecal scores, days with diarrhea, and days treated with antibiotics were increased with feeding variable amount of milk replacer over the 56-d study. Starter intake from d 1 to 56 was reduced from 919 to 717 g/d in calves fed fixed and variable amounts of milk replacer, respectively. Morbidity, measured as the number of days that calves had diarrhea, was increased by 53% when a variable amount of milk replacer was fed. Calves fed variable milk replacer were treated with antibiotics for 3.1 d compared with 1.9 d for calves fed 454 g of milk replacer/d. Concentrations of plasma glucose, urea N, and insulin-like growth factor-I were increased when calves were fed variable amount of milk replacer. Dietary supplement containing bovine serum had no effect on any parameter measured. There was no effect of milk replacer feeding on concentrations of nonesterified fatty acids, total protein, or growth hormone concentrations. Plasma tumor necrosis factor-alpha was highest in calves with the highest plasma IgG concentrations on the day of arrival and might be related to the calf's ability to identify pathogens in the environment. Under conditions of this study, calves fed variable amount of milk replacer and exposed to immunological challenge before weaning had greater BW gain, but also increased incidence of diarrhea that required added veterinary treatments. PMID- 16357285 TI - Effects of physically effective fiber on chewing activity and ruminal pH of dairy cows fed diets based on barley silage. AB - The objective of this study was to investigate the effects of physically effective neutral detergent fiber (peNDF) content of dairy cow diets containing barley silage as the sole forage source on feed intake, chewing activity, and ruminal pH. The experiment was designed as a replicated 3 x 3 Latin square using 6 lactating dairy cows with ruminal cannulas. Cows were offered 1 of 3 diets (high, medium, and low peNDF) obtained using barley silage that varied in particle length: long (theoretical cut length of 9.5 mm), medium (equal proportions of long and fine silages), and fine (theoretical cut length of 4.8 mm). The peNDF contents were determined using the Penn State Particle Separator and were 13.8, 11.8, and 10.5%, for the high, medium, and low diets, respectively. The physical effectiveness factors (defined as proportion retained on 19- and 8-mm screens) for the long and fine silages were 0.84 and 0.68, respectively. Increased forage particle size increased intake of peNDF but did not affect intake of DM and NDF. Ruminating and total chewing time were linearly increased with increasing dietary peNDF. Mean ruminal pH, area between the curve and a horizontal line drawn at pH 5.8 or 5.5, and time that pH was below 5.8 or 5.5 were not affected by peNDF content. Intake of peNDF was not correlated to any chewing activity but proportion of long particles on the 19-mm sieve tended to be correlated to ruminating chews (r = 0.36) and ruminating time (r = 0.36). These results indicate that increasing the peNDF content of diets increases chewing time. However, increased chewing time does not always improve ruminal pH status. Increasing chewing time and thus increasing salivary secretion may not fully overcome the effects of feed digestion and the production of fermentation acids that lower rumen pH. The results suggest that dietary peNDF and fermentable OM intake are critical in regulating rumen pH. Dietary particle size, expressed as peNDF, was a reliable indication of chewing activity. PMID- 16357286 TI - Effects of mixing on drinking and competitive behavior of dairy calves. AB - Group housing provides increased access to space and social interactions for calves while reducing labor costs for producers. However, group housing necessarily requires that calves be mixed and no research to date has addressed the effects of mixing on behavior of milk-fed dairy calves. The objective of this study was to monitor the feeding and competitive behavior of individual dairy calves (n = 8) after introduction into an established group of older calves fed ad libitum by a computer-controlled milk feeder. Milk feeding was monitored for 2 d before introduction into the new group and both milk feeding and competitive behaviors were monitored for 4 d after mixing. Mean (+/- SE) milk consumption before mixing was 9.7 +/- 0.7 kg/d, dropped slightly on the day of mixing to 8.6 +/- 0.6 kg/d, but increased on d 1 to 3 after mixing to 11.1 +/- 0.3 kg/d. Calves visited the feeder less frequently on the day of mixing (6.0 +/- 1.8 visits/d) than on either the days before mixing (20.3 +/- 2.5 visits/d) or the days after mixing (25.3 +/- 6.9 visits/ d). The mean duration of feeder visits and mean milk consumption per visit increased from 4 min 15 s +/- 21 s and 0.53 +/- 0.06 kg per visit before mixing to 8 min 17 s +/- 1 min 28 s and 1.87 +/- 0.51 kg per visit on the day of mixing. Competitive displacements from the milk-feeding stall were rare. In summary, feeding behavior of young calves is altered on the day of mixing, but calves are able to maintain milk intake when using a milk feeder fitted with a stall that prevents calves from displacing one another. PMID- 16357287 TI - Milk and cheese from cows fed calcium salts of palm and fish oil alone or in combination with soybean products. AB - Twenty cows were used in a randomized block design experiment for 6 wk to determine the influence of feeding partial ruminally inert Ca salts of palm and fish oil (Ca-PFO), alone or in combination with extruded full-fat soybeans or soybean oil, on milk fatty acid (FA) methyl esters composition and consumer acceptability of milk and Cheddar cheese. Cows were fed either a diet containing 44% forage and 56% concentrate (control) or a diet supplemented with 2.7% Ca-PFO (FO), 5% extruded full-fat soybeans + 2.7% Ca-PFO (FOESM), or 0.75% soybean oil + 2.7% Ca-PFO (FOSO). Total dietary FA content in the control, FO, FOESM, and FOSO diets were 4.61, 6.28, 6.77, and 6.62 g/100 g, respectively. There was no difference in nutrient intake, milk yield, or milk composition among treatments. Conjugated linoleic acid (CLA) C(18:2) cis-9, trans-11 isomer, C(18:1) trans-11 (VA), and total n-3 FA in milk from cows on the control, FO, FOESM, and FOSO treatments were 0.56, 1.20, 1.36, and 1.74; 3.29, 4.66, 6.34, and 7.81; 0.62, 0.69, 0.69, and 0.67 g/100 g of FA, respectively. Concentrations of CLA, VA, and total n-3 FA in cheese were similar to milk. A trained sensory panel detected no difference in flavors of milk and cheese, except for acid flavor below a slightly perceptible level in cheese from all treatments. Results suggest that feeding Ca PFO alone or in combination with extruded full-fat soybeans or soybean oil enhanced the CLA, VA, total unsaturated and n-3 FA in milk and cheese without negatively affecting cow performance and consumer acceptability characteristics of milk and cheese. PMID- 16357288 TI - Milk production of dairy cows fed differing concentrations of rumen-degraded protein. AB - Thirty-two multiparous and 16 primiparous Holstein cows in midlactation averaging 126 d in milk were used to determine the effects of rumen-degraded protein (RDP) concentration on lactation performance. Cows were assigned to diets in a repeated Latin square design with 3-wk experimental periods. Diets were formulated to provide 4 concentrations of dietary RDP [6.8, 8.2, 9.6, and 11.0% of dry matter (DM)] while rumen-undegraded protein remained constant (5.8% of DM). Diets contained 50% corn silage and 50% concentrate (DM basis). Ingredients within diets were equal across treatments except for ground corn, soybean meal, and ruminally protected soybean meal. Dry matter intake was not affected by treatment. Milk yield, fat yield, and protein yield all increased linearly when cows were fed diets with greater RDP. Milk fat and protein concentration each increased by 0.16 percentage units for cows fed 11% RDP compared with 6.8% RDP. Milk protein yield increased by 0.19 g/d for every 1 g/d increase in crude protein supplied mainly as RDP. As RDP increased, the efficiency of N use declined linearly. Milk urea N increased linearly when cows were fed increasing amounts of RDP, indicating increased losses of N via urine. Feeding deficient RDP diets to dairy cows can decrease nitrogen excretion, but it also decreases lactation performance. These data show an environmental benefit from underfeeding RDP to dairy cows according to National Research Council requirements, but at a financial cost to the dairy producer. PMID- 16357289 TI - Direct-fed microbial supplementation on ruminal digestion, health, and performance of pre- and postpartum dairy cattle. AB - Effects of supplementing direct-fed microbial agents (DFM) to dairy cows during the transition period were evaluated. Forty-four Holstein cows were fed close-up and lactating diets that did or did not contain 2 g of DFM/cow per d. The direct fed microbial (DFM) supplement was fed at a rate of 2 * 109 viable yeast cells and 5 * 109 cfu of bacteria per cow per day [corrected].Supplemented cows were fed the DFM 21 d prior to expected calving date through 10 wk postpartum. Cows supplemented with DFM had higher estimated ruminally available dry matter (DM) for both corn silage and haylage than did control cows. Supplemented cows consumed more DM during both the pre- and postpartum periods. In addition, those supplemented with DFM produced 2.3 kg more milk/cow per d than did nonsupplemented cows. There was no difference in 3.5% fat-corrected milk. Milk fat percentage was lower, but not depressed (4.76 vs. 4.44%) for cows receiving DFM. There were no differences in milk fat yield or milk protein percentage and yield. Cows consuming DFM had higher blood glucose postpartum, as well as lower beta-hydroxybutyrate levels both prepartum and on d 1 postpartum. Plasma nonesterified fatty acid concentration was not statistically affected by DFM, but was numerically lower prepartum and higher postpartum for supplemented cows. This study demonstrated that targeted DFM supplementation enhanced ruminal digestion of forage DM. Early lactation cows receiving supplemental DFM produced more milk and consumed more DM during the pre- and postpartum periods. Cows consuming DFM, however, experienced a lower, but not depressed, fat percentage compared with nonsupplemented cows. PMID- 16357290 TI - Feeding lactose to increase ruminal butyrate and the metabolic status of transition dairy cows. AB - Twenty-four multiparous Holstein cows (775 +/- 24 kg body weight; 3.4 +/- 0.11 body condition score) were used in a randomized complete block design experiment to determine the impact of increased ruminal butyrate from the fermentation of lactose on metabolism and lactation. Dietary treatments were either a corn-based control diet (CON) or a diet containing lactose at 15.7% of diet dry matter (LAC). Experimental diets were fed from 21 d before expected calving through 21 d in milk (DIM). Blood was sampled at -21, -14, -7, -2, 2, 7, 14, and 21 DIM, rumen fluid at -21, -7, and 7 DIM, and liver tissue via biopsy at 7 and 14 DIM. Pre- and postpartum dry matter intake (DMI) through 28 DIM averaged 12.8 and 17.7 kg/d, respectively, and did not differ between treatments; however, cows fed LAC did not exhibit a prepartum decrease in DMI. Milk yield was unaffected by treatments and averaged 45.7 kg/d during the first 70 DIM. Plasma glucose, insulin, and non-esterified fatty acids were not affected by dietary treatments. Feeding LAC increased the ruminal proportion of butyrate both pre- (11.3 vs. 9.2 +/- 0.45%) and postpartum (13.0 vs. 10.3 +/- 0.67%). Likewise, circulating plasma beta-hydroxybutyrate was increased both pre- (6.1 vs. 4.2 +/- 0.31 mg/dL) and postpartum (14.6 vs. 8.34 +/- 1.7 mg/dL) when feeding LAC compared with CON. Liver lipid content was decreased (8.6. vs. 14.7 +/- 1.5% of wet weight) in cows fed LAC relative to those fed CON, whereas liver glycogen was not affected by dietary treatments. Feeding lactose to transition dairy cows increased the proportion of butyrate in the rumen and beta-hydroxybutyrate in plasma and decreased liver lipid but did not affect lactation performance. PMID- 16357291 TI - Supplementation with extruded linseed cake affects concentrations of conjugated linoleic acid and vaccenic acid in goat milk. AB - The aim of this research was to determine the effect of adding extruded linseed cake to the dry diet of goats on the concentrations of conjugated linoleic acid (CLA) and vaccenic acid (VA) in milk fat. Thirty crossbreed dairy goats were divided into 3 groups. Their diet was supplemented with 0% (control group), 5% (low group), or 10% (high group) of extruded linseed cake (ELC), which supplied 0, 16, and 32 g/d of linseed fat, respectively. The milk fat percentage (overall mean 3.5%) and yield did not differ with the different diets, but fatty acid composition was affected by the ELC supplements. The inclusion of ELC in the diets did not influence the concentration of fatty acids from C6:0 to C12:0. The concentrations of C14:0 and C16:0 decreased as the quantity of ELC supplements increased. The concentrations (mg/100 mg of total fatty acid methyl esters) of VA (0.70, 1.23, and 1.39 in control, low, and high groups respectively) and cis 9,trans-11 CLA (0.63, 0.96, and 1.05 in control, low, and high groups, respectively) were increased by ELC supplements. The milk fat content of VA and cis- 9,trans-11 CLA were closely correlated (R2 = 0.82). Desaturation of VA in the mammary gland to produce cis-9,trans-11 CLA was higher in the control group than in the groups with ELC diets. Extruded linseed cake supplementation to lactating goats may enhance the nutritional profile of milk lipids. PMID- 16357292 TI - Milk from forage as affected by carbohydrate source and degradability with alfalfa silage-based diets. AB - Milk from forage (MF) is an estimation of the milk produced solely from forage intake. It is calculated by subtracting milk production theoretically allowed by concentrates from total milk production, assuming that maintenance requirements are covered by the forage portion of the diet. Eight multiparous Holstein cows in early lactation were used in a replicated 4 x 4 Latin square design to evaluate the impact on MF of different sources of carbohydrate with forage that was high in RDP. Diets were alfalfa-based total mixed rations that were formulated to provide similar concentrations of NEL and CP while differing in rumen degradability of concentrate carbohydrates. Treatments were 1) cracked corn (control), 2) ground corn (GC), 3) GC plus wheat starch (GC+S), and 4) GC plus dried whey permeate (GC+W). The GC and the GC+S treatments increased MF as calculated on a protein basis (14.8 vs. 10.5 kg) and increased average MF production (8.6 vs. 5.5 kg) compared with the control. Protein of forage was used more efficiently with GC and with GC+S, as shown by the lower differences between allowable MF, which estimates the potential for milk production from forage, and MF on a protein basis for these 2 treatments when compared with the control. Compared with the control, DMI increased with GC and GC+S; GC+W yielded the highest DMI. Milk production with GC+W (35.8 kg/d) was lower than with GC and GC+S (37.5 kg/d) but was higher than the control (34.0 kg/d). Milk fat concentration was higher with GC+W and lower with GC+S; GC and the control had intermediate values. Milk urea was higher with the control diet compared with the other 3 treatments. Results emphasize the advantage of using concentrates of higher degradability in the rumen to improve MF and milk production when feeding silage with high rumen-degradable protein. PMID- 16357293 TI - Development and evaluation of models to predict the feed intake of dairy cows in early lactation. AB - Inaccurate prediction of dry matter intake (DMI) limits the ability of current models to anticipate the technical and economic consequences of adopting different strategies for production management on individual dairy farms. The objective of the present study was to develop an accurate, robust, and broadly applicable prediction model and to compare it with the current NRC model for dairy cows in early lactation. Among various functions, an exponential model was selected for its best fit to DMI data of dairy cows in early lactation. Daily DMI data (n = 8,547) for 3 groups of Holstein cows (at Illinois, New Hampshire, and Pennsylvania) were used in this study. Cows at Illinois and New Hampshire were fed totally mixed diets for the first 70 d of lactation. At Pennsylvania, data were for the first 63 d postpartum. Data from Illinois cows were used as the developmental dataset, and the other 2 datasets were used for model evaluation and validation. Data for BW, milk yield, and milk composition were only available for Illinois and New Hampshire cows; therefore, only these 2 datasets were used for model comparisons. The exponential model, fitted to the individual cow daily DMI data, explained an average of 74% of the total variation in daily DMI for Illinois data, 49% of the variation for New Hampshire data, 67% of the variation for Pennsylvania data, and 64% of the variation overall. Based on all model selection criteria used in this study, the exponential model for prediction of weekly DMI of individual cows was superior to the current NRC equation. The exponential model explained 85% of the variation in weekly mean DMI compared with 42% for the NRC equation. Compared with the relative prediction error of 6% for the exponential model, that associated with prediction using the NRC equation was 14%. The overall mean square prediction error value for individual cows was 5 fold higher for the NRC equation than for the exponential model (10.4 vs. 2.0 kg2/d2). The consistently accurate and robust prediction of DMI by the exponential model for all data-sets suggested that it could safely be used for predicting DMI in many circumstances. PMID- 16357294 TI - Effects of inbreeding in the dam on dystocia and stillbirths in US Holsteins. AB - Dystocia scores were recorded by producers on 120,434 Holsteins (218,213 records) from 1985 through 1996; dystocia scores 3 to 5 were coded as difficult births. Stillbirths were recorded for deaths within the first 48 h after birth. Data were restricted to registered cows for pedigree completeness, and inbreeding coefficients were calculated using 5-generation pedigrees. Computational restrictions required that subsets of the data be created by choosing herds at random but using all records from selected herds. Effects of inbreeding in the dam were estimated in a sire-maternal grandsire (of the calf) threshold model using Gibbs sampling. The model included fixed effects of calf sex and inbreeding of the dam and random effects of herd-year-season of birth, additive genetic, and residual effects. First, second, and third parities were analyzed separately. Solutions for sex of calf and inbreeding from different parities were converted to expected change in probability of dystocia or stillbirth per 1% increase in inbreeding. Inbreeding effects were largest for first-parity cows giving birth to male calves at a 0.42% increase in probability of dystocia/1% increase in inbreeding. Effects of inbreeding for first-parity dams giving birth to female calves were smaller, 0.30%/1% increase in inbreeding. Incidence of stillbirths increased 0.25 and 0.20% for male and female calves/1% increase in inbreeding for first parity births. Effects of inbreeding on dystocia and stillbirths declined with parity. Effects of inbreeding were small, especially in later parities, but were consistently unfavorable. PMID- 16357295 TI - Wrong and missing sire information affects genetic gain in the Angeln dairy cattle population. AB - In the present study, molecular genetic markers were used to help estimate the degree of wrong sire information in the German Angeln dairy cattle population. Sixteen polymorphic microsatellite markers were genotyped on 5 different paternal half-sib families with a total of 805 daughters. For the genotyping process, blood samples of the daughters and semen samples of the sires were used. Allelic frequencies and exclusion probabilities were estimated. The simultaneous effect of wrong (WSI) and missing sire information (MSI) on the reliability of estimated breeding values and on the genetic gain was investigated using deterministic simulations. For these simulations, different values for the number of daughters per sire, heritability, WSI, and MSI were chosen. The estimated proportion of the WSI was 7% in the German Angeln dairy cattle population. The combined impact of WSI and MSI on the genetic gain was relatively large, especially in the case of small progeny size per sire and low heritability. The impact of WSI was more harmful than MSI on response to selection. PMID- 16357296 TI - Genetic aspects of growth of Holstein-Friesian dairy cows from birth to maturity. AB - In general, genetic selection is applied after first calving to traits that manifest themselves during the animal's productive life, mostly during the early part of productive life. This selection policy has had undesirable correlated responses in other economically important traits, such as health and fertility, and may also have had an effect on the growth of animals both during productive life and before first calving. In this study, we analyzed the growth trajectory of dairy heifers that had been selected for maximum production of combined fat and protein (measured in kg; select line) or for average production (control line) in the United Kingdom. Before first calving, these divergent lines were managed as a single group. Select line heifers grew faster than did control line heifers. They were also heavier at first calving, but by the end of 3 lactations, the lines were not significantly different in live weight. Selection primarily for yield and for other traits has led to heifers that grow faster and reach higher growth rates earlier in life. A genetic analysis of birth, weaning, and calving weights yielded heritability estimates of 0.53 (birth weight), 0.45 (weaning weight), and 0.75 (calving weight). Confidence intervals for the genetic correlations between the traits indicated that these BW traits are not under the same genetic control. PMID- 16357297 TI - Genetic analysis of clinical mastitis data from on-farm management software using threshold models. AB - Producer-recorded clinical mastitis data from 77,791 cows in 418 herds were used to determine the potential for genetic improvement of mastitis resistance using data from on-farm management software programs. The following threshold sire models were applied: 1) a single-trait lactation model, where mastitis was recorded as 0 or 1 in first lactation only; 2) a 3-trait lactation model, where mastitis was recorded as 0 or 1 in each of the first 3 lactations, and 3) a 12 trait, lactation-segment model, where mastitis was recorded as 0 or 1 in each of 4 segments (0 to 50, 51 to 155, 156 to 260, and 261 to 365 d postpartum) in each of the first 3 lactations. Lactation incidence rates were 0.16, 0.20, and 0.24 in first, second, and third lactation, respectively, and incidence rates within various segments of these lactations ranged from 0.036 in late first lactation to 0.093 in early third lactation. Estimated heritability of liability to clinical mastitis ranged from 0.07 to 0.15, depending on the model and stage of lactation. Heritability estimates were higher in first lactation than in subsequent lactations, but estimates were generally similar for different segments of the same lactation. Genetic correlations between lactations from the 3-trait model ranged from 0.42 to 0.49, while correlations between segments within lactation from the 12-trait model ranged from 0.26 to 0.64. Based on the results presented herein, it appears that at least 2 segments are needed per lactation, because mastitis in early lactation is lowly correlated with mastitis in mid or late lactation. Predicted transmitting abilities of sires ranged from 0.77 to 0.89 for probability of no mastitis during the first lactation and from 0.36 to 0.59 for probability of no mastitis during the first 3 lactations. Overall, this study shows that farmer-recorded clinical mastitis data can make a valuable contribution to genetic selection programs, but additional systems for gathering and storing this information must be developed, and more extensive data recording in progeny test herds should be encouraged. PMID- 16357298 TI - Performance and feeding behavior of primiparous cows loose housed alone or together with multiparous cows. AB - Lactating Holstein cows (52 multiparous and 90 primiparous) were monitored over a period of 10 mo to observe effects of grouping primiparous cows (PPC) separately from multiparous cows (MPC) on performance, feeding behavior, feed intake, feed efficiency, and milk production of PPC. Cows were kept in 2 symmetrical pens each equipped with a robotic milking unit, 2 waterers, and 28 feeding spaces. Typically, 100 lactating cows were present at a time, thereby ensuring 1.78 cows per feeding place in each pen. One pen (PP) was composed exclusively of PPC whereas the other pen (PM) included 30% PPC and 70% MPC. Primiparous cows were evenly distributed to each pen by days in milk and daily milk production. As they calved, additional primiparous cows were assigned sequentially to each of the 2 treatment groups; multiparous cows calving during the study were allocated to the PM group. Both PP and PM groups were managed equally and were fed the same basal ration twice daily plus 3 kg/d of concentrate during milking. Individual eating behavior and feed consumption at each visit were monitored electronically. Milk production was recorded daily, and milk composition monthly. Observed arithmetic means and standard errors are presented but application to other management situations is limited because animals within pen were not independent. Total dry matter intake (18.7 vs. 18.1 +/- 0.9 kg/d) and milk production (25.9 vs. 25.6 +/- 0.8 kg/d) of PPC were similar in both the PM and PP groups, respectively. Primiparous cows in the PP group had numerically more visits to the robotic milking unit (3.26 vs. 2.68 +/- 0.15) and to the feed troughs (4.91 vs. 4.02 +/- 0.43), but apparently spent less time eating (2.72 vs. 3.22 +/- 0.1 h/d) than did PPC in the PM group. Differences in feed efficiency were low but PPC in the PP group had numerically higher feed efficiency at times through 200 d in milk. Alternative grouping strategies illustrate potentially important differential responses among primiparous cows that warrant further study. PMID- 16357299 TI - Students' perception of a discussion-driven classroom environment in an upper level ruminant nutrition course with small enrollment. AB - This 3-yr classroom research project studied students' perception of the learning environment when class time was reserved primarily for discussion of preassigned reading material as an alternative to a lecture in ruminant nutrition. A student assessment-of-learning-gain instrument with 46 items was administered anonymously as a survey 3-wk into the semester and as a course evaluation at the end of the semester. Scores, collected on a scale of 1 to 10, were analyzed to determine differences between survey and evaluation, the variation due to student cohort (years 2003, 2004, and 2005), and student standing (undergraduate or graduate) for 5 selected items: item 1 = My level of interest/ curiosity for nutrition is; item 17 = The reading assignments help me learn; item 18 = The in-class discussions help me learn; item 24 = I would prefer the professor lecture; item 38 = I am learning a lot in this class. Although students indicated a preference to using class time for discussion rather than lecture, the degree of consensus was low, as 10% of scores indicated a strong desire for lecture, whereas another 10% indicated strong opposition to lecture. Reading assignments and in-class discussions contributed positively to the perception of learning and both were correlated positively with level of interest in the topic. In the evaluation, the desire for lecture was correlated negatively with level of interest in the topic and the self-assessed level of learning. Students reported a greater interest in the topic at the end of the semester than at wk 3; however, the self-reported learning gains from discussion-related activities varied with student cohort. Graduate students reported a higher interest, more learning from discussion related activities, and less desire for lecture than undergraduates. Although graduate students were more impervious to the discussion format, undergraduates perceived more learning if the discussion was student-centered (i.e., focused on their questions and answers), rather than subject-centered (i.e., focused on the paper content). PMID- 16357300 TI - Prediction and evaluation of urine and urinary nitrogen and mineral excretion from dairy cattle. AB - Urine excretion is a substantial factor in the amount of manure that needs to be managed, and urinary N can contribute to ammonia volatilization. Development and validation of prediction equations focusing on dietary factors to decrease urine and urinary nutrient excretion will provide information for managing urine and feces separately or for other future technologies. The objective of this study was to develop equations for prediction of urine excretion and excretion of urinary N, Na, and K and to evaluate both new and previously published prediction equations for estimation of urine and urinary nutrient excretion from lactating dairy cows. Data sets from metabolism studies conducted at Washington State University were compiled and evaluated for excretion of minerals. Urine excretion averaged 24.1 kg/d and urinary nitrogen excretion ranged from 63 to 499 g/d in the calibration data set. Regression equations were developed to predict urine excretion, urinary N excretion, and urinary Na and K excretion. Predictors used in the regression equations included milk yield, body weight, dietary crude protein percentage, milk urea nitrogen, and nutrient intakes. Previously published prediction equations were evaluated using data sets from Washington State University and the University of Wisconsin. Mean and linear biases were evaluated by determining the regression of residuals on predicted values. Evaluation and validation of prediction equations are important to develop equations that will more accurately estimate urine and urinary nitrogen excretion from lactating dairy cows. PMID- 16357301 TI - Trends in calving ages and calving intervals for dairy cattle breeds in the United States. AB - Trends since 1980 for calving age and calving interval, 2 factors that influence herd life, were examined by parity for 5 breeds of US dairy cattle. Calving data were from cows with records that passed edits for USDA genetic evaluations and were in herds that remained on Dairy Herd Improvement test. First-calf heifers calved at progressively younger ages over time, but the age decline was less for later parities because of longer calving intervals. Breed differences for calving age were evident for all parities; current mean age at first calving ranged from 24 mo for Jerseys to 28 mo for Ayrshires. Mean calving age across all parities declined over time for all breeds, primarily because of increased turnover rate, and ranged from 48 mo for Holsteins to 54 mo for Ayrshires. Across parity, annual increase in calving interval was reasonably consistent (0.90 to 1.07 d/yr) for all breeds except Jersey (0.49 d/yr). Within parity, regressions of calving interval on year were generally similar to overall breed trend. Breed means for first calving interval across time ranged from 390 d for Jerseys to 407 d for Brown Swiss. PMID- 16357302 TI - Selective Rac-1 inhibition protects from diabetes-induced vascular injury. AB - Diabetes mellitus is a main risk factor for vascular diseases. Vascular injury induced by diabetes mellitus is characterized by endothelial dysfunction attributable to an increased oxidative stress. So far, the molecular mechanisms involved in the vasculotoxic effects of diabetes are only partially known. We examined the effect of diabetes mellitus on oxidative stress and Rac-1 activation, a small G-protein involved in the activation of NADPH oxidase. Our results show that oxidative stress in vessels of different murine models of diabetes mellitus and in endothelial cells treated with high glucose is associated with an increased Rac-1/PAK binding and Rac-1 translocation from cytosol to plasma membrane, thus demonstrating an enhanced Rac-1 activity. More important, selective Rac-1 inhibition by an adenoviral vector carrying a dominant negative mutant of Rac-1 protected from oxidative stress and vascular dysfunction induced by diabetes mellitus. Our study demonstrates that Rac-1 plays a crucial role in diabetes-induced vascular injury, and it could be a target of novel therapeutic approaches to reduce vascular risk in diabetes mellitus. PMID- 16357303 TI - Downregulation of endothelin-1 by farnesoid X receptor in vascular endothelial cells. AB - The farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily that is highly expressed in liver, kidney, adrenals, and intestine. FXR may play an important role in the pathogenesis of cardiovascular diseases via regulating the metabolism and transport of cholesterol. In this study, we report that FXR is also expressed in rat pulmonary artery endothelial cells (EC), a "nonclassical" bile acid target tissue. FXR is functional in EC, as demonstrated by induction of its target genes such as small heterodimer partner (SHP) after treatment with chenodeoxycholic acid, a FXR agonist. Interestingly, activation of FXR in EC led to downregulation of endothelin (ET)-1 expression. Reporter assays showed that activation of FXR inhibited transcriptional activation of the human ET-1 gene promoter and also repressed the activity of a heterologous promoter driven by activator protein (AP)-1 response elements. Electrophoretic mobility-shift and chromatin immunoprecipitation assays indicated that FXR reduced the binding activity of AP-1 transcriptional factors, suggesting that FXR may suppress ET-1 expression via negatively interfering with AP-1 signaling. These studies suggest that FXR may play a role in endothelial homeostasis and may serve as a novel molecular target for manipulating ET-1 expression in vascular EC. PMID- 16357304 TI - Xanthine oxidoreductase inhibition causes reverse remodeling in rats with dilated cardiomyopathy. AB - Increased reactive oxygen species (ROS) generation is implicated in cardiac remodeling in heart failure (HF). As xanthine oxidoreductase (XOR) is 1 of the major sources of ROS, we tested whether XOR inhibition could improve cardiac performance and induce reverse remodeling in a model of established HF, the spontaneously hypertensive/HF (SHHF) rat. We randomized Wistar Kyoto (WKY, controls, 18 to 21 months) and SHHF (19 to 21 months) rats to oxypurinol (1 mmol/L; n=4 and n=15, respectively) or placebo (n=3 and n=10, respectively) orally for 4 weeks. At baseline, SHHF rats had decreased fractional shortening (FS) (31+/-3% versus 67+/-3% in WKY, P<0.0001) and increased left-ventricular (LV) end-diastolic dimension (9.7+/-0.2 mm versus 7.0+/-0.4 mm in WKY, P<0.0001). Whereas placebo and oxypurinol did not change cardiac architecture in WKY, oxypurinol attenuated decreased FS and elevated LV end-diastolic dimension, LV end-systolic dimension, and LV mass in SHHF. Increased myocyte width in SHHF was reduced by oxypurinol. Additionally, fetal gene activation, altered calcium cycling proteins, and upregulated phospho-extracellular signal-regulated kinase were restored toward normal by oxypurinol (P<0.05 versus placebo-SHHF). Importantly, SHHF rats exhibited increased XOR mRNA expression and activity, and oxypurinol treatment reduced XOR activity and superoxide production toward normal, but not expression. On the other hand, NADPH oxidase activity remained unchanged, despite elevated subunit protein abundance in treated and untreated SHHF rats. Together these data demonstrate that chronic XOR inhibition restores cardiac structure and function and offsets alterations in fetal gene expression/Ca2+ handling pathways, supporting the idea that inhibiting XOR derived oxidative stress substantially improves the HF phenotype. PMID- 16357305 TI - Bone morphogenetic protein receptor-2 signaling promotes pulmonary arterial endothelial cell survival: implications for loss-of-function mutations in the pathogenesis of pulmonary hypertension. AB - Mutations in the bone morphogenetic protein (BMP) receptor-2 (BMPR2) have been found in patients with idiopathic pulmonary arterial hypertension (IPAH); however, the mechanistic link between loss of BMPR2 signaling and the development of pulmonary arterial hypertension is unclear. We hypothesized that, contrary to smooth muscle cells, this pathway promotes survival in pulmonary artery endothelial cells (ECs) and loss of BMPR2 signaling will predispose to EC apoptosis. ECs were treated with BMP-2 or BMP-7 (200 ng/mL) for 24 hours in regular or serum-free (SF) medium, with and without addition of tumor necrosis factor alpha, and apoptosis was assessed by flow cytometry (Annexin V), TUNEL, or caspase-3 activity. Treatment for 24 hours in SF medium increased apoptosis, and both BMP-2 and BMP-7 significantly reduced apoptosis in response to serum deprivation to levels not different from serum controls. Transfection with 5 microg of small interfering RNAs for BMPR2 produced specific gene silencing assessed by RT-PCR and Western blot analysis. BMPR2 gene silencing increased apoptosis almost 3-fold (P=0.0027), even in the presence of serum. Circulating endothelial progenitor cells (EPCs) isolated from normal subjects or patients with IPAH were differentiated in culture for 7 days and apoptosis was determined in the presence and absence of BMPs. BMP-2 reduced apoptosis induced by serum withdrawal in EPCs from normal subjects but not in EPCs isolated from patients with IPAH. These results support the hypothesis that loss-of-function mutations in BMPR2 could lead to increased pulmonary EC apoptosis, representing a possible initiating mechanism in the pathogenesis of pulmonary arterial hypertension. PMID- 16357306 TI - Functional TRPM7 channels accumulate at the plasma membrane in response to fluid flow. AB - Many cells are constantly exposed to fluid mechanical forces generated by flowing blood, and wall shear stresses modulate aspects of their structure and function. However, the mechanisms for mechanotransduction of flow are not well understood. Here we report that TRPM7, which is both an ion channel and a functional kinase, is translocated within cells in response to laminar flow. After exposure of cells to physiological values of laminar fluid flow, the number of TRPM7 molecules localized at or near the plasma membrane increased up to 2-fold, in less than 100 seconds. This increase in membrane-localized GFP-TRPM7, as seen by total internal reflection fluorescence microscopy, closely correlated with increases in TRPM7 current. Both endogenous and heterologously expressed TRPM7 was found in tubulovesicular structures that were translocated to the region of the plasma membrane on induction of shear stress. In vascular smooth muscle cells, but not in several types of endothelial cells, fluid flow increased endogenous native TRPM7 current amplitude. We hypothesize that TRPM7 plays a role in pathological response to vessel wall injury. PMID- 16357307 TI - Compartmentalized phosphodiesterase-2 activity blunts beta-adrenergic cardiac inotropy via an NO/cGMP-dependent pathway. AB - beta-Adrenergic signaling via cAMP generation and PKA activation mediates the positive inotropic effect of catecholamines on heart cells. Given the large diversity of protein kinase A targets within cardiac cells, a precisely regulated and confined activity of such signaling pathway is essential for specificity of response. Phosphodiesterases (PDEs) are the only route for degrading cAMP and are thus poised to regulate intracellular cAMP gradients. Their spatial confinement to discrete compartments and functional coupling to individual receptors provides an efficient way to control local [cAMP]i in a stimulus-specific manner. By performing real-time imaging of cyclic nucleotides in living ventriculocytes we identify a prominent role of PDE2 in selectively shaping the cAMP response to catecholamines via a pathway involving beta3-adrenergic receptors, NO generation and cGMP production. In cardiac myocytes, PDE2, being tightly coupled to the pool of adenylyl cyclases activated by beta-adrenergic receptor stimulation, coordinates cGMP and cAMP signaling in a novel feedback control loop of the beta adrenergic pathway. In this, activation of beta3-adrenergic receptors counteracts cAMP generation obtained via stimulation of beta1/beta2-adrenoceptors. Our study illustrates the key role of compartmentalized PDE2 in the control of catecholamine-generated cAMP and furthers our understanding of localized cAMP signaling. PMID- 16357308 TI - Destabilization and early termination of spiral-wave reentry by a class III antiarrhythmic agent, nifekalant, in a perfused two-dimensional layer of rabbit ventricular myocardium. PMID- 16357309 TI - Glycoprotein Ibalpha-mediated platelet adhesion and aggregation to immobilized thrombin under conditions of flow. AB - OBJECTIVE: Thrombin interacts with platelets via the protease-activated receptors (PARs) 1 and 4, and via glycoprotein Ibalpha (GPIbalpha). Recently, it was shown that platelets are able to adhere to immobilized thrombin under static conditions via GPIbalpha. METHODS AND RESULTS: Here, we show that platelets are also able to adhere to and form stable aggregates on immobilized thrombin under conditions of flow. Adhesion and aggregation to thrombin was dependent on the interaction with GPIbalpha, as addition of glycocalicin or an antibody blocking the interaction between thrombin and GPIbalpha inhibited platelet adhesion. Additionally, platelet adhesion to recombinant thrombin mutants, which are unable to bind GPIbalpha, was severely suppressed. Furthermore, platelet adhesion to thrombin was dependent on activation of PARs, and partly on granule secretion and thromboxane-A2 synthesis. Immobilization of thrombin on a fibrin network resulted in substantially increased adhesion compared with fibrin alone. The adhesion to fibrin alone was completely abolished by addition of dRGDW, whereas fibrin-bound thrombin still showed substantial platelet adhesion in the presence of dRGDW, indicating that fibrin-bound thrombin is able to directly capture platelets under flow. CONCLUSIONS: These results indicate that platelets are able to adhere to thrombin under flow conditions, which is dependent on the interaction with GPIbalpha. PMID- 16357310 TI - Reduced immunoregulatory CD31+ T cells in patients with atherosclerotic abdominal aortic aneurysm. AB - BACKGROUND: Cell-mediated immunity is considered to contribute to the pathogenesis of abdominal aortic aneurysms (AAA). In particular, infiltrating macrophages and CD8+ T lymphocytes participate in the destruction of the aortic wall extracellular matrix and smooth muscle cells. We surmise that these pathological events are controlled by circulating regulatory lymphocytes. METHODS AND RESULTS: Circulating CD4+/CD31+ cells were reduced in AAA patients (n=80, 8.9+/-0.6%) as compared with controls (n=69, 13.7+/-0.8%; P<0.001) and inversely proportional to AAA size. Exclusion of the aneurysm by an endoprothesis did not affect CD31+ T cell values. Reduction of blood CD4+/CD31+ cells was not attributable to their enrichment in AAA tissue. In contrast, CD8+/CD31+ cells were slightly reduced in the blood while increased in the aneurysmal tissue (29.2+/-0.5 versus 20.2+/-4.7% in blood, n=6; P<0.05). Remarkably, high percentages of CD4+/CD31+ cells were able to regulate proliferation and cytokine production of CD8+ lymphocytes, as well as CD8+ cell-mediated cytotoxicity of aortic smooth muscle cells (P<0.01). Finally, CD4+/CD31+ cells reduced the production and activity of metalloproteinase-9 by lipopolysaccharide-stimulated macrophages. CONCLUSIONS: Circulating CD4+/CD31+ T cells regulate macrophage and CD8+ T cell activation and effector function in the arterial wall. Their reduction might promote the development of AAA. PMID- 16357311 TI - Elevated concentrations of nonesterified fatty acids increase monocyte expression of CD11b and adhesion to endothelial cells. AB - OBJECTIVE: Monocyte proinflammatory activity has been demonstrated in obesity, insulin resistance, and type 2 diabetes, metabolic conditions that are frequently associated with elevated levels of nonesterified fatty acids (NEFA). We therefore tested the hypothesis that NEFA may induce monocyte inflammation. METHODS AND RESULTS: Monocytes exposed to NEFA for 2 days demonstrated a dose-related increase in intracellular reactive oxygen species (ROS) formation and adhesion to endothelial cells. All of these effects were inhibited by the coaddition of antioxidants such as glutathione or butylated hydroxytoluene, by inhibition of ROS generation by NADPH oxidase inhibitors, and by inhibition of protein kinase C, a recognized stimulator of NAPDH oxidase. Monocytes exposed to NEFA also demonstrated a significant increase in CD11b message expression. Stimulation of monocyte adhesion to endothelial cells by NEFA was inhibited by addition of neutralizing antibodies to either CD11b or CD18. Finally, surface expression of CD11b increased significantly on monocytes as measured by flow cytometry, after their incubation with NEFA. CONCLUSIONS: These studies indicate that elevated concentrations of NEFA may enhance integrin facilitated monocyte adhesion to endothelial cells and these effects appear mediated, in part, through activation of NADPH oxidase and oxidative stress. PMID- 16357312 TI - Effects of rosiglitazone on lipids, adipokines, and inflammatory markers in nondiabetic patients with low high-density lipoprotein cholesterol and metabolic syndrome. AB - BACKGROUND: PPAR-gamma agonists improve insulin sensitivity and glycemic control in type 2 diabetes and may reduce atherosclerosis progression. Thus, PPAR-gamma agonists may be an effective therapy for metabolic syndrome. However, the full spectrum of potentially antiatherogenic mechanisms of PPAR-gamma agonists have not been fully tested in nondiabetic patients with metabolic syndrome. METHODS AND RESULTS: We performed a prospective, double-blinded, placebo-controlled study of 60 nondiabetic subjects with low high-density lipoprotein cholesterol (HDL-C) level and metabolic syndrome to rosiglitazone 8 mg daily or placebo for 12 weeks. We found no significant effect of rosiglitazone on HDL-C (+5.5% versus +5.8%, P=0.89), and an increase in total cholesterol (+8% versus -1%; P=0.03). Nevertheless, rosiglitazone significantly increased adiponectin (+168% versus +25%; P<0.001), and lowered resistin (-6% versus +4%; P=0.009), C-reactive protein (-32% versus +36%, P=0.002), interleukin (IL)-6 (-22% versus +4%, P<0.001), and soluble tumor-necrosis factor-alpha receptor-2 (-5% versus +7%, P<0.001). CONCLUSIONS: These findings suggest that rosiglitazone, presumably through its PPAR-gamma agonist properties, has direct effects on inflammatory markers and adipokines in the absence of favorable lipid effects. These findings may help explain the mechanism underlying the possible antiatherosclerotic effects of rosiglitazone. PMID- 16357313 TI - Antioxidants inhibit the ability of lysophosphatidylcholine to regulate proteoglycan synthesis. AB - OBJECTIVE: We previously have shown that lysophosphatidylcholine (lysoPC) regulates proteoglycan synthesis by vascular smooth muscle cells (SMCs). Given the accumulating evidence for reactive oxygen species (ROS) as mediators of a variety of effects of lysoPC, the present study evaluates the potential role of ROS as intermediate molecules in the regulation of proteoglycan synthesis by lysoPC. METHODS AND RESULTS: LysoPC (10 micromol/L) was found to stimulate rapid and sustained generation of ROS by SMC, as indicated using a fluorescent probe for measuring intracellular oxidants and fluorescence-activated cell sorting. This was not associated with cytotoxicity, as evaluated by fluorescence microscopy using MitoTracker Red or propidium iodide, cell number, cell protein, or lactate dehydrogenase release. Pretreatment with catalase or superoxide dismutase, specific scavengers of hydrogen peroxide and superoxide, respectively, blocked the ability of lysoPC to stimulate both accumulation of ROS and proteoglycan synthesis. Most importantly, these enzymatic antioxidants prevented lysoPC from stimulating the synthesis of proteoglycans with enhanced lipoprotein binding properties, as quantified by a gel shift binding assay. CONCLUSIONS: These findings strongly suggest that ROS are key mediators in the ability of lysoPC to regulate proteoglycan synthesis and that these effects can be inhibited by antioxidants. PMID- 16357314 TI - 18F-choline images murine atherosclerotic plaques ex vivo. AB - OBJECTIVE: Current imaging modalities of atherosclerosis mainly visualize plaque morphology. Valuable insight into plaque biology was achieved by visualizing enhanced metabolism in plaque-derived macrophages using 18F-fluorodeoxyglucose (18F-FDG). Similarly, enhanced uptake of 18F-fluorocholine (18F-FCH) was associated with macrophages surrounding an abscess. As macrophages are important determinants of plaque vulnerability, we tested 18F-FCH for plaque imaging. METHODS AND RESULTS: We injected 18F-FCH (n=5) or 18F-FDG (n=5) intravenously into atherosclerotic apolipoprotein E-deficient mice. En face measurements of aortae isolated 20 minutes after 18F-FCH injections demonstrated an excellent correlation between fat stainings and autoradiographies (r=0.842, P<0.0001), achieving a sensitivity of 84% to detect plaques by 18F-FCH. In contrast, radiotracer uptake 20 minutes after 18F-FDG injections correlated less with en face fat stainings (r=0.261, P<0.05), reaching a sensitivity of 64%. Histological analyses of cross-sections 20 minutes after coinjections of 18F-FCH and 14C-FDG (n=3) showed that 18F-FCH uptake correlated better with fat staining (r=0.740, P<0.0001) and macrophage-positive areas (r=0.740, P<0.0001) than 14C-FDG (fat: r=0.236, P=0.29 and CD68 staining: r=0.352, P=0.11), respectively. CONCLUSIONS: 18F-FCH identifies murine plaques better than 18F-FDG using ex vivo imaging. Enhanced 18F-FCH uptake into macrophages may render this tracer a promising candidate for imaging plaques in patients. PMID- 16357315 TI - Occupation, marital status, and low-grade inflammation: mutual confounding or independent cardiovascular risk factors? AB - OBJECTIVE: We explored the relationships between inflammatory proteins, occupation, and marital status, and their independent associations with incidence of cardiovascular disease (CVD). METHODS AND RESULTS: Five inflammation-sensitive proteins (ISPs) (fibrinogen, ceruloplasmin, haptoglobin, alpha1-antitrypsin, orosomucoid) were measured in 6075 apparently healthy men. Incidence of coronary events and stroke was followed over 18 years in relation to occupation and marital status. All ISPs showed higher concentrations in divorced men and in manual workers. Except for fibrinogen, this remained significant after adjustments for confounding factors. Adjusted for traditional cardiovascular risk factors, incidence of coronary events was significantly increased in unskilled manual workers and in divorced men. The relative risks were slightly reduced after further adjustments for ISPs (from 1.79 to 1.70 in unskilled manual workers; from 1.58 to 1.51 in divorced men). All ISPs were significantly associated with incidence of coronary events, after adjustments for traditional risk factors. This relationship was essentially unchanged after further adjustments for occupation and marital status. CONCLUSIONS: Inflammation could contribute to, but not fully explain, the increased cardiovascular risk in manual workers and divorced men. Although the ISPs vary greatly by occupational and marital status, this does not confound the relationship between ISPs and incidence of CVD. PMID- 16357316 TI - Effect of vectors on human endothelial cell signal transduction: implications for cardiovascular gene therapy. AB - OBJECTIVE: Endothelium is an important target for gene therapy. We have investigated the effect of viral and nonviral vectors on the phenotype and function of endothelial cells (ECs) and developed methods to block any activation caused by these vectors. METHODS AND RESULTS: Transduction of ECs with viral vectors, including adenovirus, lentiviruses, and Moloney murine leukemia virus, can induce a pro-inflammatory phenotype. This activation was reduced when nonviral vectors were used. We demonstrate that after transduction there is upregulation of dsRNA-triggered antiviral and PI3K/Akt signaling pathway. Blockade of the NFkappaB, PI3-K, or PKR signaling pathways all operated to inhibit partially virally induced activation, and inhibition of both PKR and PI3 K pathways totally blocked EC activation. Furthermore, inhibition of IFN alpha/beta in addition to PI3-K was effective at preventing EC activation. CONCLUSIONS: Viral vectors, although efficient at transducing ECs, result in their activation. Blockade of the signaling pathways involved in viral activation may be used to prevent such activation. PMID- 16357317 TI - ABCA1 and ABCG1 synergize to mediate cholesterol export to apoA-I. AB - OBJECTIVE: To study the acceptor specificity for human ABCG1 (hABCG1)-mediated cholesterol efflux. METHODS AND RESULTS: Cells overexpressing hABCG1 were created in Chinese Hamster Ovary (CHO-K1) cells and characterized in terms of lipid composition. hABCG1 expressed in these cells formed homodimers and was mostly present intracellularly. Cholesterol efflux from hABCG1 cells to HDL2 and HDL3 was increased but not to lipid-free apolipoproteins. A range of phospholipid containing acceptors apart from high-density lipoprotein (HDL) subclasses were also efficient in mediating ABCG1-dependent export of cholesterol. Importantly, a buoyant phospholipid-containing fraction generated from incubation of lipid-free apoA-I with macrophages was nearly as efficient as HDL2. The capacity of acceptors to induce ABCG1-mediated efflux was strongly correlated with their total phospholipid content, suggesting that acceptor phospholipids drive ABCG1 mediated efflux. Most importantly, acceptors for ABCG1-mediated cholesterol export could be generated from incubation of cells with lipid-free apoA-I through the action of ABCA1 alone. CONCLUSIONS: These results indicate a synergistic relationship between ABCA1 and ABCG1 in peripheral tissues, where ABCA1 lipidates any lipid-poor/free apoA-I to generate nascent or pre-beta-HDL. These particles in turn may serve as substrates for ABCG1-mediated cholesterol export. PMID- 16357318 TI - Endothelial ephrinB2 is controlled by microenvironmental determinants and associates context-dependently with CD31. AB - OBJECTIVE: The EphB ligand ephrinB2 has been identified as a critical determinant of arterial endothelial differentiation and as a positive regulator of invading endothelial cells during angiogenesis. This study was aimed at identifying determinants of endothelial cell ephrinB2 expression. METHODS AND RESULTS: Arteriovenous asymmetrical endothelial cell ephrinB2 expression in vivo is lost on transfer into culture with aortic endothelial cells becoming partially ephrinB2-negative and saphenous vein endothelial cells becoming partially ephrinB2-positive. Contact with smooth muscle cells and angiogenic stimulation by vascular endothelial growth factor lead to an increased endothelial cell ephrinB2 expression. Quiescent, smooth muscle-contacting endothelial cells express ephrinB2 uniformly on their luminal surface. In contrast, monolayer endothelial cells translocate ephrinB2 to interendothelial cell junctions, which is strongly enhanced by EphB4-Fc-mediated receptor body activation. Junctional ephrinB2 colocalizes and coimmunoprecipitates with CD31. CONCLUSIONS: This study identifies distinct regulatory mechanisms of endothelial ephrinB2 expression and cellular distribution in quiescent and activated endothelial cells. The data demonstrate that endothelial cell ephrinB2 expression is controlled by microenvironmental determinants rather than being an intrinsic endothelial cell differentiation marker. PMID- 16357319 TI - A negative carotid plaque area test is superior to other noninvasive atherosclerosis studies for reducing the likelihood of having underlying significant coronary artery disease. AB - OBJECTIVE: Coronary calcium score (CCS), carotid plaque area (CPA), intima-media thickness (IMT), and C-reactive protein (CRP) are independent predictors of cardiovascular prognosis. Although each test may enhance risk stratification, their comparative abilities to screen for underlying coronary stenoses in individual patients is less established. METHODS AND RESULTS: Forty-two patients who had a 16-slice coronary computed tomography angiogram (CTA) performed were invited to have CPA, IMT, and CRP measured. CPA was defined as the sum of all the cross-sectional areas of each plaque >1 mm in diameter found in all carotid vessels bilaterally. CCS and the number plus degree of stenotic coronary arteries were determined by CTA. The presence of clinically significant coronary artery disease (CAD) was defined as the existence of any stenosis > or =50%. CTA identified clinically significant CAD in 43% of the patients. CPA >0 was more sensitive (72%) and specific (58%) than a CCS >0 (58% and 55%) for identifying CAD. A "clean" carotid artery (CPA=0) provides a superior negative predictive value (74%) and likelihood ratio of a negative test (0.48) than all other studies, in particular versus a CCS=0 (65% and 0.72). The areas under the receiver-operator curves for CPA and CCS in relation to any CAD were similar (0.640 versus 0.675). Carotid IMT and CRP performed poorly compared with CPA and CCS. For detecting CAD in only the left main or left anterior descending artery, the negative predictive value and likelihood ratio of a negative test remained superior for CPA (87% and 0.33) compared with CCS (80% and 0.56). In our population with a prevalence of these coronary lesions of 30%, the post-test probability in any patient with a negative CPA result is reduced to 10%. CONCLUSIONS: CPA determination is superior to CCS, IMT, and CRP in its ability to reduce the likelihood of clinically significant underlying CAD in patients of varying cardiac risk. PMID- 16357320 TI - Selective activation of TACI by syndecan-2. AB - B-lymphocyte homeostasis and function are regulated by complementary actions of the TNFR family members TACI, BCMA, and BAFF-R, which are expressed by mature B cells. How these receptors are differentially activated is not entirely understood, because the primary ligand BAFF binds to all three. We searched for alternative ligands for TACI using recombinant TACI-Fc fusion protein as a probe and identified syndecan-2 as a new binding partner. TACI binding appears to require heparan sulfate posttranslational modifications of syndecan-2, because free heparin or pretreatment with heparitinase blocked the interaction. Syndecan 2 bound TACI but bound neither BAFF-R nor BCMA. Transfected cells expressing syndecan-2 activated signaling through TACI, as indicated by an NFAT-specific reporter. Syndecan-1 and syndecan-4 were also able to induce TACI signaling in a similar manner. This is the first identification of ligands that selectively activate TACI without simultaneously triggering BCMA or BAFF-R. This finding may help explain the alternative outcomes of signaling from this family of receptors in B cells. PMID- 16357321 TI - Ectodermally derived steel/stem cell factor functions non-cell autonomously during primitive erythropoiesis in Xenopus. AB - Signals derived from nonhematopoietic tissues are essential for normal primitive erythropoiesis in vertebrates, but little is known about the nature of these signals. In Xenopus, unidentified factors secreted by ectodermal cells during gastrulation are required to enable the underlying ventral mesoderm to form blood. Steel is expressed in the ectoderm of early Xenopus embryos and is known to regulate definitive erythroid progenitor survival and differentiation in other organisms, making it an excellent candidate regulator of primitive erythropoiesis. In this study, we tested whether steel signaling is required for primitive red blood cell differentiation in mice and frogs. We show that Xsl is expressed in the ectoderm in Xenopus gastrulae and that c-kit homologs are expressed in the underlying mesoderm at the same stages of development. We present loss of function data in whole Xenopus embryos and explants that demonstrate a requirement for ectodermally derived steel to signal through c-kit in the mesoderm to support early steps in the differentiation of primitive erythroid but not myeloid cells. Finally, we show that primitive erythropoiesis is not disrupted in mouse embryos that lack c-kit function. Our data suggest a previously unrecognized and unique function of steel/c-kit during primitive erythropoiesis in Xenopus. PMID- 16357322 TI - Altered IL-7Ralpha expression with aging and the potential implications of IL-7 therapy on CD8+ T-cell immune responses. AB - We investigated the effects of aging on the IL-7-mediated CD8+ T-cell survival pathway and of IL-7 therapy on T-cell immunity. Cells expressing IL-7 receptor (IL-7R) alphahigh and alphalow were identified in a CD45RA+ effector memory (EM(CD45RA+), CD45RA+CCR7-) CD8+ T-cell subset. Elderly subjects (65 years and older) had an increased frequency of EM(CD45RA+) IL-7Ralphalow) CD8+ T cells, leading to decreased STAT5 phosphorylation and survival responses to IL-7 compared with young subjects (40 years and younger). These EM(CD45RA+) IL 7Ralphalow cells were largely antigen experienced (CD27-CD28-), replicatively senescent (CD57+), and perforinhigh CD8+ T cells that had decreased IL-7Ralpha mRNA, independent of guanine and adenine binding protein alpha (GABPalpha) and growth factor independence-1 (GFI1) expression. In measuring T-cell receptor (TCR) repertoires of EM(CD45RA+) CD8+ T cells, the elderly had a limited repertoire in IL-7Ralphahigh and IL-7Ralphalow cells, whereas the young had a diverse repertoire in IL-7Ralphahigh but not in IL-7Ralphalow cells. These findings suggest that aging affects IL-7Ralpha expression by EM(CD45RA+) CD8+ T cells, leading to impaired signaling and survival responses to IL-7, and that IL 7 therapy may improve the survival of EM(CD45RA+) CD8+ T cells with a diverse TCR repertoire in the young but not in the elderly. PMID- 16357323 TI - T-bet concomitantly controls migration, survival, and effector functions during the development of Valpha14i NKT cells. AB - Valpha14i natural killer T (NKT)-cell function has been implicated in a number of disease conditions. The molecular events that drive Valpha14i NKT-cell development remain elusive. We recently showed that T-bet is required for the terminal maturation of these cells. Here we identify some of the genetic targets of T-bet during Valpha14i NKT-cell lineage development. Microarray gene expression analyses on developing Valpha14i NKT cells were performed and provide a molecular framework to study these maturation events. In vitro ectopic expression of T-bet in immature Valpha14i NKT cells, which do not yet express T bet, was sufficient to promote Valpha14i NKT-cell maturation, driving the expression of multiple genes, including those that participate in migration, survival, and effector functions. By regulating the expression of T-helper 1 (Th1)-associated cytokines, chemokines, chemokine receptors, and molecules involved in cytolysis, T-bet defines the unique lineage attributes of mature Valpha14i NKT cells and acts to link these attributes to a developmental process. PMID- 16357324 TI - The small GTPase Rap1b regulates the cross talk between platelet integrin alpha2beta1 and integrin alphaIIbbeta3. AB - The involvement of the small GTPase Rap1b in platelet integrin alpha2beta1 dependent outside-in signaling was investigated. Platelet adhesion to 4 different specific ligands for integrin alpha2beta1, monomeric collagen, decorin, and collagen-derived peptides CB8(II) and CB11(II), induced a robust and rapid activation of Rap1b. This process did not require secreted ADP or thromboxane A2 production but was critically regulated by phospholipase C (PLC)-derived second messengers. Both Ca2+ and protein kinase C were found to organize independent but additive pathways for Rap1b activation downstream of integrin-alpha2beta1, which were completely blocked by inhibition of PLC with U73122. Moreover, integrin alpha2beta1 engagement failed to trigger Rap1b activation in murine platelets lacking CalDAG-GEFI, a guanine nucleotide exchange factor regulated by Ca2+ and diacylglycerol, despite normal phosphorylation and activation of PLCgamma2. In addition, CalDAG-GEFI-deficient platelets showed defective integrin alpha2beta1 dependent adhesion and spreading. We found that outside-in signaling through integrin alpha2beta1 triggered inside-out activation of integrin alphaIIbbeta3 and promoted fibrinogen binding. Similarly to Rap1b stimulation, this process occurred downstream of PLC activation and was dramatically impaired in murine platelets lacking the Rap1 exchange factor CalDAG-GEFI. These results demonstrate that Rap1b is an important element in integrin-dependent outside-in signaling during platelet adhesion and regulates the cross talk between adhesive receptors. PMID- 16357325 TI - Novel stabilin-1 interacting chitinase-like protein (SI-CLP) is up-regulated in alternatively activated macrophages and secreted via lysosomal pathway. AB - Mammalian Glyco_18-domain-containing proteins include catalytically active chitinases and chitinase-like proteins with cytokine activity involved in host defense and Th2-type inflammatory reactions. Here, we describe a novel human Glyco_18-domain-containing protein, SI-CLP, as an interacting partner of the endocytic/sorting receptor stabilin-1. Similarly to the chitinase-like cytokines YKL-39, YKL-40, and YM1/2, SI-CLP lacks a chitin-binding domain and catalytic amino acids. Using a novel mAb 1C11, we demonstrated that SI-CLP is sorted into late endosomes and secretory lysosomes in human alternatively activated macrophages. The direct interaction of SI-CLP with stabilin-1, their colocalization in the trans-Golgi network, and the reduced sorting of SI-CLP into lysosomes in macrophages treated with stabilin-1 siRNA suggest that stabilin-1 is involved in intracellular sorting of SI-CLP. Expression of SI-CLP in macrophages was strongly up-regulated by the Th2 cytokine IL-4 and by dexamethasone. This effect was suppressed by IFNgamma but not affected by IL-10. In contrast, expression of YKL-40 was induced by IFNgamma and suppressed by dexamethasone. Macrophages treated with IL-4 secreted SI-CLP, while costimulation with dexamethasone blocked secretion and resulted in intracellular accumulation of SI CLP. The 1C11 mAb detected SI-CLP in human bronchoalveolar lavage and peripheral blood leukocytes (PBLs), and can be used to analyze the role of SI-CLP in human disorders. PMID- 16357326 TI - Human mast cells express multiple EP receptors for prostaglandin E2 that differentially modulate activation responses. AB - Prostaglandin E2 (PGE2) blocks mast-cell (MC)-dependent allergic responses in humans but activates MCs in vitro. We assessed the functions of the EP receptors for PGE2 on cultured human MCs (hMCs). hMCs expressed the EP3, EP2, and EP4 receptors. PGE2 stimulated the accumulation of cyclic adenosine monophosphate (cAMP), and suppressed both Fc epsilonRI-mediated eicosanoid production and tumor necrosis factor-alpha (TNF-alpha) generation. PGE2 also caused phosphorylation of extracellular signal-regulated kinase (ERK), exocytosis, and production of prostaglandin D2 (PGD2), as well as leukotriene C4 (LTC4) when protein kinase A (PKA) was inhibited. An EP3 receptor-selective agonist, AE-248, mimicked PGE2 mediated ERK phosphorylation, exocytosis, and eicosanoid formation. Selective agonists of both EP2 and EP4 receptors (AE1-259-01 and AE-329, respectively) stimulated cAMP accumulation. No selective agonist, alone or in combination, was as effective as PGE2. AE-248, AE1-259-01, and AE-329 all inhibited Fc epsilonRI mediated TNF-alpha generation, while AE1-259-01 blocked eicosanoid production. PGE2 caused the expression of inducible cAMP early repressor (ICER) by a pathway involving PKA and ERK. Thus, while PGE2 activates MCs through EP3 receptors, it also counteracts Fc epsilonRI-mediated eicosanoid production through EP2 receptors and PKA, and blocks cytokine transcription. These functions explain the potency of PGE2 as a suppressor of early- and late-phase allergic responses. PMID- 16357328 TI - The Notch ligand delta-1 is a hematopoietic development cofactor for plasmacytoid dendritic cells. AB - Plasmacytoid dendritic cells (pDCs) play an important role in innate and adaptive immunity, prompting interest in mechanisms controlling the production of this lineage of cells. Notch signaling via one of the Notch ligands, delta-like 1 (delta-1), influences the hematopoietic development of several lymphoid and myeloid lineages, but whether or not delta-1 affects the formation of pDCs is unknown and was tested here. Human CD34+ progenitor cells were cultured onto delta-1-expressing OP9 stroma in the presence of flt-3 ligand and IL-7, and this efficiently generated BDCA-2+ CD123+ CD4+ CD11c- cells with the characteristic morphology of pDCs, expressing toll-like receptor-9 (TLR9), pre-Talpha mRNAs, and secreting CpG-induced IFN-alpha. Delta-1 augmented the numbers of BDCA-2+ cells produced without affecting their proliferation, and the effect was blocked by gamma-secretase inhibition. The development of pDCs was stroma-, delta-1-, and cytokine-dependent and could be induced from committed lymphoid progenitor cells, which responded to delta-1 by opposite changes in pDC- and B-cell production. Our results identify delta-1 as a novel factor enhancing pDC hematopoiesis and delineate a new role for Notch signaling in lymphopoiesis by showing its opposite effect on pDC and B lineage determination. PMID- 16357327 TI - Lack of alpha4 integrin expression in stem cells restricts competitive function and self-renewal activity. AB - Alpha4 integrin or VLA4 (CD49d/CD29) is a multitask molecule with wide expression within and outside the hematopoietic system. Because targeted ablation of alpha4 integrin leads to embryonic lethality, to study its effects on adult hematopoiesis, we used animals with conditional excision of alpha4 integrin (alpha4Delta/Delta) in hematopoietic cells. In such animals, we previously documented weakened bone marrow retention of progenitor cells during homeostasis and impaired homing and short-term engraftment after transplantation. In the present study we show that long-term repopulating cells lacking alpha4 integrins display a competitive disadvantage in hematopoietic reconstitution compared to normal competitors. Although initial dominance of alpha4+ competitors is due to their better homing and proliferative expansion early after transplantation, a progressive decline in contribution of alpha4Delta/Delta hematopoiesis is compatible with neither normal homing nor normal function of alpha4Delta/Delta hematopoietic stem cells (HSCs) in post-homing hematopoiesis. In the absence of alpha4+ competitor cells, alpha4Delta/Delta HSCs can establish long-term hematopoiesis in primary recipients, however, some resurgence of host hematopoiesis is evident, and it becomes dominant in secondary transplants, so that no survivors with exclusively alpha4Delta/Delta cells are seen in tertiary transplants. Collectively, our data provide compelling evidence that under regenerative stress alpha4 integrin assumes a greater importance than for maintenance of steady-state hematopoiesis. PMID- 16357329 TI - CD40-activated B cells express full lymph node homing triad and induce T-cell chemotaxis: potential as cellular adjuvants. AB - CD40-activated B cells (CD40-B cells) have previously been introduced as an alternative source of antigen-presenting cells for immunotherapy. CD40-B cells can prime naive and expand memory T cells, and they can be generated in large numbers from very small amounts of peripheral blood derived from healthy individuals or cancer patients alike. Administration of CD40-B cells as a cellular adjuvant would require these cells to migrate toward secondary lymphoid organs and attract T cells in situ, processes guided by specific chemokines and chemokine receptors. Here, we demonstrate that primary, human CD40-B cells express a pattern of adhesion molecules and chemokine receptors necessary for homing to secondary lymphoid organs and have the capacity to migrate to cognate ligands. Furthermore, we show that CD40-B cells express important T-cell attractants and induce strong T-cell chemotaxis. These findings further support the use of CD40-B cells as cellular adjuvant for cancer immunotherapy. PMID- 16357330 TI - Additional minocycline pleurodesis after thoracoscopic surgery for primary spontaneous pneumothorax. AB - RATIONALE: Ipsilateral recurrence rates of spontaneous pneumothorax after video assisted thoracoscopic surgery are higher than rates after open thoracotomy. OBJECTIVES: This study was conducted to determine whether additional minocycline pleurodesis would be effective in diminishing recurrence after video-assisted thoracoscopic surgery treatment of primary spontaneous pneumothorax. METHODS: Between June 2001 and February 2004, 202 patients with primary spontaneous pneumothorax were treated by conventional or needlescopic video-assisted thoracoscopic surgery. The procedures included resection of blebs and mechanical pleurodesis by scrubbing the parietal pleura. After the operation, patients were randomly assigned to additional minocycline pleurodesis (103 patients) or to observation (99 patients). MAIN RESULTS: Patients in the minocycline group had higher intensity chest pain and required a higher accumulated dose of meperidine. Short-term results showed that the two groups had comparable chest drainage duration, postoperative hospital stay, and complication rates. Patients in the minocycline group demonstrated a trend of decreased rate of prolonged air leaks (1.9 vs. 6.1%, p = 0.100). After a mean follow-up of 29 mo (12-47 mo), recurrent ipsilateral pneumothorax was noted in two patients in the minocycline group and eight patients in the observation group (p = 0.044 by the Kaplan-Meier method and log-rank test). Postoperative long-term residual chest pain and pulmonary function were comparable in both groups. CONCLUSIONS: Although associated with intense immediate chest pain, additional minocycline pleurodesis is a safe and convenient procedure that can reduce the rate of ipsilateral recurrence after thoracoscopic treatment for primary spontaneous pneumothorax. PMID- 16357331 TI - Effects of esomeprazole 40 mg twice daily on asthma: a randomized placebo controlled trial. AB - RATIONALE: Gastroesophageal reflux disease (GERD) is common in patients with asthma, suggesting an interaction between the two conditions. OBJECTIVES: To assess the effect of gastric acid suppression with the proton pump inhibitor esomeprazole on asthma outcomes in subjects with persistent moderate to severe asthma treated with antiinflammatory asthma medication. METHODS: In this double blind study, subjects were randomized to receive esomeprazole 40 mg or placebo twice daily for 16 wk. According to nocturnal respiratory symptoms (NOC) and GERD, subjects were divided into three strata: GERD-/NOC+, GERD+/NOC-, and GERD+/NOC+. MEASUREMENTS AND MAIN RESULTS: A total of 770 subjects were randomized. There was no statistically significant improvement in morning peak expiratory flow (PEF) over placebo in the overall study population: 6.3 L/min (p = 0.061). Over the whole treatment period, in GERD+/NOC+ subjects (n = 350), esomeprazole provided an 8.7-L/min improvement (p = 0.03) in morning PEF, and a 10.2-L/min improvement (p = 0.012) in evening PEF over placebo. Among 307 subjects taking long-acting beta2-agonists, improvements over placebo were observed in morning PEF (12.2 L/min, p = 0.017) and in evening PEF (11.1 L/min, p = 0.024); these improvements were more pronounced in GERD+/NOC+ subjects. Esomeprazole 40 mg twice daily was well tolerated and no safety concerns were noted. CONCLUSIONS: Esomeprazole improved PEF in subjects with asthma who presented with both GERD and NOC. In subjects without both GERD and NOC, no improvement could be detected. PMID- 16357332 TI - Resident pleural macrophages are key orchestrators of neutrophil recruitment in pleural inflammation. AB - RATIONALE: The role played by resident pleural macrophages in the initiation of pleural inflammation is currently unclear. OBJECTIVE: To evaluate the role of resident pleural macrophages in the initiation of inflammation. METHODS: We have used a conditional macrophage ablation strategy to determine the role of resident pleural macrophages in the regulation of neutrophil recruitment in a murine model of experimental pleurisy induced by the administration of carrageenan and formalin- fixed Staphylococcus aureus. MEASUREMENTS AND MAIN RESULTS: Conditional macrophage ablation mice express the human diphtheria toxin receptor under the control of the CD11b promoter such that the administration of diphtheria toxin induces ablation of nearly 97% of resident macrophages. Ablation of resident pleural macrophages before the administration of carrageenan or S. aureus dramatically reduced neutrophil influx into the pleural cavity. In the carrageenan model, the reduction in neutrophil infiltration was associated with marked early reduction in the level of macrophage inflammatory protein 2 as well as reduced levels of various cytokines, including tumor necrosis factor alpha, interleukin 6, and interleukin 10. Adoptive transfer of nontransgenic macrophages partially restored neutrophil infiltration. We also stimulated macrophage depleted and nondepleted pleural cell populations with carrageenan in vitro and determined the production of chemokines and cytokines. Chemokine and cytokine production was markedly reduced by macrophage depletion, reinforcing the role of resident pleural macrophages in the generation of mediators that initiate acute inflammation. CONCLUSION: These studies indicate a critical role for resident pleural macrophages in sensing perturbation to the local microenvironment and orchestrating subsequent neutrophil infiltration. PMID- 16357334 TI - In vivo evidence for radial migration of neurons by long-distance somal translocation in the developing ferret visual cortex. AB - During the development of the cerebral cortex, neurons generated in the cortical ventricular zone migrate radially toward the marginal zone. Radially migrating neurons are thought to display 1 of 2 morphologies: cells with a long, pia contacting, apical process utilized for somal translocation early in development, when the cortex is still relatively thin; or cells with a short leading process, abundant at late stages of corticogenesis when neurons need to travel for longer distances. In large convoluted brains, like those of many primates and carnivores, radially migrating neurons must travel distances up to several millimeters before reaching their final destination, often following curvilinear trajectories. Here we analyze modes and morphologies of radially migrating neurons in convoluted brains by studying the visual cortex of developing ferrets. We provide in vivo and in vitro evidence for the existence of late-born cortical neurons that migrate radially by long-distance somal translocation within a long apical process extended to the cortical plate, in contrast to the early somal translocation observed in rodents. Long-distance translocating neurons in the ferret show a discontinuous rhythm of migration, alternating periods of advance with periods of stall. Furthermore, by combining different labeling methods we find the simultaneous presence in the developing ferret cortex of long-distance translocating neurons and neurons migrating within a short leading process. PMID- 16357333 TI - Neural system for controlling the contents of object working memory in humans. AB - Working memory (WM), the active maintenance of currently relevant information, is a flexible system allowing for fast and frequent goal-directed changes of rehearsed information. Successful WM maintenance prevents interference from distracting stimuli while allowing new task-relevant information to update the contents of WM. We used functional magnetic resonance imaging to show that when WM contents were updated, regardless of stimulus type (faces or houses), a frontoparietal network showed transient increases in activation. Some of these regions are highly similar to those identified in studies of shifting attention, supporting the idea that updating WM involves a change in the attentional priority afforded to the current perceptual input. A region within the mid ventrolateral prefrontal cortex, near the junction of the inferior frontal sulcus and precentral sulcus (inferior frontal junction), that has previously been implicated in cognitive control, demonstrated transient increases in activity during updating as well as sustained maintenance activity. A more anterior prefrontal region, middle frontal gyrus, previously implicated in protecting the contents of WM from interfering stimuli during maintenance, demonstrated transient increases in activity during updating. The current study suggests that updating WM results from a combination of increased attention to the visual stimulus and a change in the system's interference protection state. PMID- 16357335 TI - The independence of memory traces of attended and unattended stimuli. AB - In a dichotic paradigm, duration-deviant mismatch negativity (MMN) largely reflects standard stimuli presented to the same ear as the deviant, suggesting an independent representation for each ear. We sought to assess this representation independence. Twenty-two participants, presented left (all 2000 Hz, 70-dB sound pressure level [SPL]) and right (all 800 Hz, 85-dB SPL) ear sounds, attended to 1 ear per block. A series of successive trains of 6-30 sounds were presented to each ear. Although 1 ear received short (40 ms) standard sounds, the other received an equal number of long (120 ms) standards. In the next train, the standard durations were switched between the ears. Stimulus onset asynchrony ranged from 120 to 440 ms. Duration deviants (240 ms) replaced the final standards of some trains. MMN latency simultaneously moved earlier for the ear changing from long to short standards and later for the ear changing from short to long standards. Based on a simple linear model, more than 80% of the memory trace reflected within-channel standards. We conclude that independently changeable memory traces underlie dichotic presentation. Separation of the representation for the attended subset of stimuli is proposed as core to the mechanism of sustained attention. PMID- 16357336 TI - Oscillatory beta activity predicts response speed during a multisensory audiovisual reaction time task: a high-density electrical mapping study. AB - Bisensory redundant targets are processed faster than the respective unisensory target stimuli alone as evidenced by substantially faster reaction times (RTs). This multisensory RT facilitation has been interpreted as an expression of integrative processing between the different sensory modalities. However, the neuronal mechanisms underlying the RT facilitation effect are not well understood. Oscillatory responses in the beta frequency range (13-30 Hz) have been related to sensory-motor processing. Here, we investigated whether modulation of beta responses might also underlie the faster RTs seen for multisensory stimuli. Using high-density electrical mapping, we explored the association between early (50-170 ms) multisensory processing in the evoked beta response and RTs recorded during a simple RT task. Subjects were instructed to indicate the appearance of any stimulus in a stream of auditory-alone (A), visual alone (V), and multisensory (AV) stimuli by a button press. Beta responses were analyzed using Morlet wavelet transformations. Multisensory interactions were found over frontal, occipital, central, and sensory-motor regions. Critically, beta activity correlated with mean RTs over all stimulus types. Significant negative correlations were found for frontal, occipital, and sensory-motor scalp regions. We conclude that the association between oscillatory beta activity and integrative multisensory processing is directly linked to multisensory RT facilitation effects. PMID- 16357337 TI - Effects of human cerebellar thalamus disruption on adaptive control of reaching. AB - Lesion or degeneration of the cerebellum can profoundly impair adaptive control of reaching in humans. Computational models have proposed that internal models that help control movements form in the cerebellum and influence planned motor output through the cerebello-thalamo-cortical pathway. However, lesion studies of the cerebellar thalamus have not consistently found impairment in reaching or adaptation of reaching. To elucidate the role of the cerebellar thalamus in humans, we studied a group of essential tremor (ET) patients with deep brain stimulation (DBS) electrodes placed in the cerebellar thalamus. The stimulation can be turned on or off remotely and is thought to reduce tremor by blocking the spread of the pathological output from the cerebellum. We studied the effect of thalamic DBS on the ability to adapt arm movements to novel force fields. Although thalamic DBS resulted in a dramatic and significant reduction of tremor in ET, it also impaired motor adaptation: the larger the stimulation voltage, the greater the reduction in rates of adaptation. We next examined ET patients that had undergone unilateral thalamotomy in the cerebellar thalamus and found that adaptation with the contralateral arm was impaired compared with the ipsilateral arm. Therefore, although both lesion and electrical stimulation of the cerebellar thalamus are highly effective in reducing tremor, they significantly impair the ability of the brain to form internal models of action. Adaptive control of reaching appears to depend on the integrity of the cerebello-thalamo-cortical pathway. PMID- 16357338 TI - Quantitative aspects of corticocortical connections: a tracer study in the mouse. AB - This study provides neuroanatomical data relevant to models and simulations of the propagation of activity over the cortex. We administered small injections of the anterograde tracer biotinylated dextran amine to various regions of the mouse cortex (1 per animal). Two-dimensional reconstructions of the cortical surface were made, showing the distribution, size, and density of the terminal fields. Within the injected hemisphere, the largest part of the terminal field always surrounded the injection site and extended over neighboring areas. On average, axons from injection sites of 0.05). Controlling for baseline performance, intervention participants prescribed more safely than controls after receiving the CDSS (0.23 vs. 0.45 [F = 4.24, p < 0.05]). With the CDSS, intervention participants documented more complete assessment of patient gastrointestinal risk from NSAIDs. CONCLUSION: PARTICIPANTS provided with a PDA-based CDSS for NSAID prescribing made fewer unsafe treatment decisions than participants without the CDSS. PMID- 16357351 TI - The effects of creating psychological ownership on physicians' acceptance of clinical information systems. AB - OBJECTIVE: Motivated by the need to push further our understanding of physicians' acceptance of clinical information systems, we propose a relatively new construct, namely, psychological ownership. We situated the construct within a nomological net using a prevailing and dominant information technology adoption behavior model as a logical starting point. DESIGN: A mail survey was sent to the population of users of a regional physician order entry (POE) system aimed at speeding up the transmission of clinical data, mainly laboratory tests and radiology examinations, within a community health network. MEASUREMENTS: All scales, but one, were measured using previously validated instruments. For its part, the psychological ownership scale was developed using a multistage iterative procedure. RESULTS: Ninety-one questionnaires were returned to the researchers, for a response rate of 72.8%. Our findings reveal that, in order to foster physicians' adoption of a clinical information system, it is important to encourage and cultivate a positive attitude toward using the new system. In this connection, positive perception of the technology's usefulness is crucial. Second, results demonstrate that psychological ownership of a POE system is positively associated with physicians' perceptions of system utility and system user friendliness. Last, through their active involvement and participation, physicians feel they have greater influence on the development process, thereby developing feelings of ownership toward the clinical system. CONCLUSION: Psychological ownership's highly significant associations with user participation and crucial beliefs driving technology acceptance behaviors among physicians affirm the value of this construct in extending our understanding of POE adoption. PMID- 16357352 TI - Reducing workload in systematic review preparation using automated citation classification. AB - OBJECTIVE: To determine whether automated classification of document citations can be useful in reducing the time spent by experts reviewing journal articles for inclusion in updating systematic reviews of drug class efficacy for treatment of disease. DESIGN: A test collection was built using the annotated reference files from 15 systematic drug class reviews. A voting perceptron-based automated citation classification system was constructed to classify each article as containing high-quality, drug class-specific evidence or not. Cross-validation experiments were performed to evaluate performance. MEASUREMENTS: Precision, recall, and F-measure were evaluated at a range of sample weightings. Work saved over sampling at 95% recall was used as the measure of value to the review process. RESULTS: A reduction in the number of articles needing manual review was found for 11 of the 15 drug review topics studied. For three of the topics, the reduction was 50% or greater. CONCLUSION: Automated document citation classification could be a useful tool in maintaining systematic reviews of the efficacy of drug therapy. Further work is needed to refine the classification system and determine the best manner to integrate the system into the production of systematic reviews. PMID- 16357353 TI - A context-sensitive approach to anonymizing spatial surveillance data: impact on outbreak detection. AB - OBJECTIVE: The use of spatially based methods and algorithms in epidemiology and surveillance presents privacy challenges for researchers and public health agencies. We describe a novel method for anonymizing individuals in public health data sets by transposing their spatial locations through a process informed by the underlying population density. Further, we measure the impact of the skew on detection of spatial clustering as measured by a spatial scanning statistic. DESIGN: Cases were emergency department (ED) visits for respiratory illness. Baseline ED visit data were injected with artificially created clusters ranging in magnitude, shape, and location. The geocoded locations were then transformed using a de-identification algorithm that accounts for the local underlying population density. MEASUREMENTS: A total of 12,600 separate weeks of case data with artificially created clusters were combined with control data and the impact on detection of spatial clustering identified by a spatial scan statistic was measured. RESULTS: The anonymization algorithm produced an expected skew of cases that resulted in high values of data set k-anonymity. De-identification that moves points an average distance of 0.25 km lowers the spatial cluster detection sensitivity by less than 4% and lowers the detection specificity less than 1%. CONCLUSION: A population-density-based Gaussian spatial blurring markedly decreases the ability to identify individuals in a data set while only slightly decreasing the performance of a standardly used outbreak detection tool. These findings suggest new approaches to anonymizing data for spatial epidemiology and surveillance. PMID- 16357354 TI - Who are the informaticians? What we know and should know. AB - The beginning of the 21st century has seen a surge in interest and enthusiasm for health care information technology based on its ability to demonstrate improvements in the quality, safety, and cost-efficiency of health care. One question, however, for which we have fewer answers is "who will be the individuals that develop, implement, and evaluate these systems?" In particular, while most attention has been paid to the exemplar leaders in health information technology, less has been focused on the issue of the workforce necessary to sustain the systems to achieve their vision. The discipline of medical informatics must pay sufficient attention to the professional workforce that will deploy systems outside the informatics research setting so their benefits may more widely accrue. PMID- 16357355 TI - MachineProse: an ontological framework for scientific assertions. AB - OBJECTIVE: The idea of testing a hypothesis is central to the practice of biomedical research. However, the results of testing a hypothesis are published mainly in the form of prose articles. Encoding the results as scientific assertions that are both human and machine readable would greatly enhance the synergistic growth and dissemination of knowledge. DESIGN: We have developed MachineProse (MP), an ontological framework for the concise specification of scientific assertions. MP is based on the idea of an assertion constituting a fundamental unit of knowledge. This is in contrast to current approaches that use discrete concept terms from domain ontologies for annotation and assertions are only inferred heuristically. MEASUREMENTS: We use illustrative examples to highlight the advantages of MP over the use of the Medical Subject Headings (MeSH) system and keywords in indexing scientific articles. RESULTS: We show how MP makes it possible to carry out semantic annotation of publications that is machine readable and allows for precise search capabilities. In addition, when used by itself, MP serves as a knowledge repository for emerging discoveries. A prototype for proof of concept has been developed that demonstrates the feasibility and novel benefits of MP. As part of the MP framework, we have created an ontology of relationship types with about 100 terms optimized for the representation of scientific assertions. CONCLUSION: MachineProse is a novel semantic framework that we believe may be used to summarize research findings, annotate biomedical publications, and support sophisticated searches. PMID- 16357356 TI - Monitoring device safety in interventional cardiology. AB - OBJECTIVE: A variety of postmarketing surveillance strategies to monitor the safety of medical devices have been supported by the U.S. Food and Drug Administration, but there are few systems to automate surveillance. Our objective was to develop a system to perform real-time monitoring of safety data using a variety of process control techniques. DESIGN: The Web-based Data Extraction and Longitudinal Time Analysis (DELTA) system imports clinical data in real-time from an electronic database and generates alerts for potentially unsafe devices or procedures. The statistical techniques used are statistical process control (SPC), logistic regression (LR), and Bayesian updating statistics (BUS). MEASUREMENTS: We selected in-patient mortality following implantation of the Cypher drug-eluting coronary stent to evaluate our system. Data from the University of Michigan Consortium Bare-Metal Stent Study was used to calculate the event rate alerting boundaries. Data analysis was performed on local catheterization data from Brigham and Women's Hospital from July 1, 2003, shortly after the Cypher release, to December 31, 2004, including 2,270 cases with 27 observed deaths. RESULTS: The single-stratum SPC had alerts in months 4 and 10. The multistrata SPC had alerts in months 5, 10, and 18 in the moderate-risk stratum, and months 1, 4, 7, and 10 in the high-risk stratum. The only cumulative alerts were in the first month for the high-risk stratum of the multistrata SPC. The LR method showed no monthly or cumulative alerts. The BUS method showed an alert in the first month for the high-risk stratum. CONCLUSION: The system performed adequately within the Brigham and Women's Hospital Intranet environment based on the design goals. All three cumulative methods agreed that the overall observed event rates were not significantly higher for the new medical device than for a closely related medical device and were consistent with the observation that the initial concerns about this device dissipated as more data accumulated. PMID- 16357357 TI - The development and evaluation of an integrated electronic prescribing and drug management system for primary care. AB - OBJECTIVE: To develop and evaluate the acceptability and use of an integrated electronic prescribing and drug management system (MOXXI) for primary care physicians. DESIGN: A 20-month follow-up study of MOXXI (Medical Office of the XXIst Century) implementation in 28 primary care physicians and 13,515 consenting patients. MEASUREMENT: MOXXI was developed to enhance patient safety by integrating patient demographics, retrieving active drugs from pharmacy systems, generating an automated problem list, and providing electronic prescription, stop order, automated prescribing problem alerts, and compliance monitoring functions. Evaluation of technical performance, acceptability, and use was conducted using audit trails, questionnaires, standardized tasks, and information from comprehensive health insurance databases. RESULTS: Perceived improvements in continuity of care and professional autonomy were associated with physicians' expected use of MOXXI. Physician speed in using MOXXI improved substantially in the first three months; however, only the represcribing function was faster using MOXXI than by handwritten prescription. Physicians wrote electronic prescriptions in 36.9 per 100 visits and reviewed the patient's drug profile in 12.6 per 100 visits. Physicians rated printed prescriptions, the current drug list, and the represcribing function as the most beneficial aspects of the system. Physicians were more likely to use the drug profile for patients who used more medication, made more emergency department visits, had more prescribing physicians, and lower continuity of care. CONCLUSION: Primary care physicians believed an integrated electronic prescribing and drug management system would improve continuity of care, and they were more likely to use the system for patients with more complex, fragmented care. PMID- 16357358 TI - Overriding of drug safety alerts in computerized physician order entry. AB - Many computerized physician order entry (CPOE) systems have integrated drug safety alerts. The authors reviewed the literature on physician response to drug safety alerts and interpreted the results using Reason's framework of accident causation. In total, 17 papers met the inclusion criteria. Drug safety alerts are overridden by clinicians in 49% to 96% of cases. Alert overriding may often be justified and adverse drug events due to overridden alerts are not always preventable. A distinction between appropriate and useful alerts should be made. The alerting system may contain error-producing conditions like low specificity, low sensitivity, unclear information content, unnecessary workflow disruptions, and unsafe and inefficient handling. These may result in active failures of the physician, like ignoring alerts, misinterpretation, and incorrect handling. Efforts to improve patient safety by increasing correct handling of drug safety alerts should focus on the error-producing conditions in software and organization. Studies on cognitive processes playing a role in overriding drug safety alerts are lacking. PMID- 16357359 TI - Using biometrics for participant identification in a research study: a case report. AB - This paper illustrates the use of biometrics through the application of an iris based biometrics system for identifying twins and their parents in a longitudinal research study. It explores the use of biometrics (science of measuring physical or anatomical characteristics of individuals) as a technology for correct identification of individuals during longitudinal studies to help ensure data fidelity. Examples of these circumstances include longitudinal epidemiological and genetic studies, clinical trials, and multicenter collaborative studies where accurate identification of subjects over time can be difficult when the subject may be young or an unreliable source of identification information. The use of technology can automate the process of subject identification thereby reducing the need to depend on subject recall during repeated visits thus helping to ensure data quality. This case report provides insights that may serve as useful hints for those responsible for planning system implementation that involves participants' authentication that would require a more secure form of identification. PMID- 16357361 TI - Fatty acid transport protein 1 and long-chain acyl coenzyme A synthetase 1 interact in adipocytes. AB - The fatty acid transport proteins (FATP) and long-chain acyl coenzyme A synthetase (ACSL) proteins have been shown to play a role in facilitating long chain fatty acid (LCFA) transport in mammalian cells under physiologic conditions. The involvement of both FATP and ACSL proteins is consistent with the model of vectorial acylation, in which fatty acid transport is coupled to esterification. This study was undertaken to determine whether the functions of these proteins are coordinated through a protein-protein interaction that might serve as a point of regulation for cellular fatty acid transport. We demonstrate for the first time that FATP1 and ACSL1 coimmunoprecipitate in 3T3-L1 adipocytes, indicating that these proteins form an oligomeric complex. The efficiency of FATP1 and ACSL1 coimmunoprecipitation is unaltered by acute insulin treatment, which stimulates fatty acid uptake, or by treatment with isoproterenol, which decreases fatty acid uptake and stimulates lipolysis. Moreover, inhibition of ACSL1 activity in adipocytes impairs fatty acid uptake, suggesting that esterification is essential for fatty acid transport. Together, our findings suggest that a constitutive interaction between FATP1 and ACSL1 contributes to the efficient cellular uptake of LCFAs in adipocytes through vectorial acylation. PMID- 16357360 TI - Integrating "best of care" protocols into clinicians' workflow via care provider order entry: impact on quality-of-care indicators for acute myocardial infarction. AB - OBJECTIVE: In the context of an inpatient care provider order entry (CPOE) system, to evaluate the impact of a decision support tool on integration of cardiology "best of care" order sets into clinicians' admission workflow, and on quality measures for the management of acute myocardial infarction (AMI) patients. DESIGN: A before-and-after study of physician orders evaluated (1) per patient use rates of standardized acute coronary syndrome (ACS) order set and (2) patient-level compliance with two individual recommendations: early aspirin ordering and beta-blocker ordering. MEASUREMENTS: The effectiveness of the intervention was evaluated for (1) all patients with ACS (suspected for AMI at the time of admission) (N = 540) and (2) the subset of the ACS patients with confirmed discharge diagnosis of AMI (n = 180) who comprise the recommended target population who should receive aspirin and/or beta-blockers. Compliance rates for use of the ACS order set, aspirin ordering, and beta-blocker ordering were calculated as the percentages of patients who had each action performed within 24 hours of admission. RESULTS: For all ACS admissions, the decision support tool significantly increased use of the ACS order set (p = 0.009). Use of the ACS order set led, within the first 24 hours of hospitalization, to a significant increase in the number of patients who received aspirin (p = 0.001) and a nonsignificant increase in the number of patients who received beta blockers (p = 0.07). Results for confirmed AMI cases demonstrated similar increases, but did not reach statistical significance. CONCLUSION: The decision support tool increased optional use of the ACS order set, but room for additional improvement exists. PMID- 16357362 TI - Caveolin-1 confers antiinflammatory effects in murine macrophages via the MKK3/p38 MAPK pathway. AB - Caveolin-1 has been reported to regulate apoptosis, lipid metabolism, and endocytosis in macrophages. In the present study, we demonstrate that caveolin-1 can act as a potent immunomodulatory molecule. We first observed caveolin-1 expression in murine alveolar macrophages by Western blotting and immunofluorescence microscopy. Loss-of-function experiments using small interfering RNA showed that down regulating caveolin-1 expression in murine alveolar and peritoneal macrophages increased LPS-induced proinflammatory cytokine TNF-alpha and IL-6 production but decreased anti-inflammatory cytokine IL-10 production. Gain-of-function experiments demonstrated that overexpression of caveolin-1 in RAW264.7 cells decreased LPS-induced TNF-alpha and IL-6 production and augmented IL-10 production. p38 mitogen-activated protein kinase (MAPK) phosphorylation was increased by overexpressing caveolin-1 in RAW264.7 cells, whereas c-Jun N-terminal kinase, extracellular signal-regulated kinase MAPK, and Akt phosphorylation were inhibited. The antiinflammatory modulation of LPS-induced cytokine production by caveolin-1 was significantly abrogated by the administration of p38 inhibitor SB203580 in RAW264.7 cells. Peritoneal macrophages isolated from MKK3 null mice did not demonstrate any modulation of LPS-induced cytokine production by caveolin-1. LPS-induced activation of NF kappaB and AP-1 determined by electrophoretic mobility shift assay were significantly reduced by overexpressing caveolin-1 in RAW264.7 cells. The reductions were attenuated by the administration of p38 inhibitor SB203580. Taken together, our data suggest that caveolin-1 acts as a potent immunomodulatory effector molecule in immune cells and that the regulation of LPS-induced cytokine production by caveolin-1 involves the MKK3/p38 MAPK pathway. PMID- 16357363 TI - P53 mediates amosite asbestos-induced alveolar epithelial cell mitochondria regulated apoptosis. AB - Asbestos causes pulmonary toxicity in part by generating reactive oxygen species that cause DNA damage. We previously showed that the mitochondria-regulated (intrinsic) death pathway mediates alveolar epithelial cell (AEC) DNA damage and apoptosis. Because p53 regulates the DNA damage response in part by inducing intrinsic cell death, we determined whether p53-dependent transcriptional activity mediates asbestos-induced AEC mitochondrial dysfunction and apoptosis. We show that inhibitors of p53-dependent transcriptional activation (pifithrin and type 16-E6 protein) block asbestos-induced AEC mitochondrial membrane potential change (DeltaPsim), caspase 9 activation, and apoptosis. We demonstrate that asbestos activates p53 promoter activity, mRNA levels, protein expression, and Bax and p53 mitochondrial translocation. Further, pifithrin, E6, phytic acid, or rho(0)-A549 cells (cells incapable of mitochondrial reactive oxygen species production) block asbestos-induced p53 activation. Finally, we show that asbestos augments p53 expression in cells at the bronchoalveolar duct junctions of rat lungs and that phytic acid prevents this. These data suggest that p53-dependent transcription pathways mediate asbestos-induced AEC mitochondria-regulated apoptosis. This suggests an important interactive effect between p53 and the mitochondria in the pathogenesis of asbestos-induced pulmonary toxicity that may have broader implications for our understanding of pulmonary fibrosis and lung cancer. PMID- 16357364 TI - Impact of mitochondria and NADPH oxidases on acute and sustained hypoxic pulmonary vasoconstriction. AB - Hypoxic pulmonary vasoconstriction (HPV) matches lung perfusion with ventilation to optimize pulmonary gas exchange. However, it remains unclear whether acute HPV (occurring within seconds) and the vasoconstrictor response to sustained alveolar hypoxia (developing over several hours) are triggered by identical mechanisms. We investigated the effect of mitochondrial and NADPH oxidase inhibitors on both phases of HPV in intact rabbit lungs. These studies revealed that the sustained HPV is largely dependent on mitochondrial complex I and totally dependent on complex IV, whereas NADPH oxidase dependence was only observed for acute HPV. These findings were reinforced by an alternative approach employing lungs from mice deficient in the NADPH oxidase subunit p 47(phox). In these mice (which lack a subunit suggested to be important for the function of most NADPH oxidase isoforms), but not in gp 91(phox)-deficient mice (which represent only one isoform of NADPH oxidases), acute HPV was significantly reduced, while non hypoxia-induced vasoconstrictions elicited by the thromboxane mimetic U46619 were not affected. We concluded that the acute phase and the sustained phase of HPV are differentially regulated, with NADPH oxidase activity predominating in the acute phase, while a strong dependence on mitochondrial participation was observed for the second phase. PMID- 16357365 TI - Hyperoxia-induced reactive oxygen species formation in pulmonary capillary endothelial cells in situ. AB - Lung capillary endothelial cells (ECs) are a critical target of oxygen toxicity and play a central role in the pathogenesis of hyperoxic lung injury. To determine mechanisms and time course of EC activation in normobaric hyperoxia, we measured endothelial concentration of reactive oxygen species (ROS) and cytosolic calcium ([Ca(2+)](i)) by in situ imaging of 2',7'-dichlorofluorescein (DCF) and fura 2 fluorescence, respectively, and translocation of the small GTPase Rac1 by immunofluorescence in isolated perfused rat lungs. Endothelial DCF fluorescence and [Ca(2+)](i) increased continuously yet reversibly during a 90-min interval of hyperoxic ventilation with 70% O(2), demonstrating progressive ROS generation and second messenger signaling. ROS formation increased exponentially with higher O(2) concentrations. ROS and [Ca(2+)](i) responses were blocked by the mitochondrial complex I inhibitor rotenone, whereas inhibitors of NAD(P)H oxidase and the intracellular Ca(2+) chelator BAPTA predominantly attenuated the late phase of the hyperoxia-induced DCF fluorescence increase after > 30 min. Rac1 translocation in lung capillary ECs was barely detectable at normoxia but was prominent after 60 min of hyperoxia and could be blocked by rotenone and BAPTA. We conclude that hyperoxia induces ROS formation in lung capillary ECs, which initially originates from the mitochondrial electron transport chain but subsequently involves activation of NAD(P)H oxidase by endothelial [Ca(2+)](i) signaling and Rac1 activation. Our findings demonstrate rapid activation of ECs by hyperoxia in situ and identify mechanisms that may be relevant in the initiation of hyperoxic lung injury. PMID- 16357367 TI - The mucin-1 568 adenosine to guanine polymorphism influences serum Krebs von den Lungen-6 levels. AB - Krebs von den Lungen (KL)-6 offers a new perspective as a disease marker in pulmonary diseases. The aim of this study was to analyze whether serum KL-6 levels are dependent on the functional adenosine to guanine mucin-1 (MUC1) gene polymorphism at nucleotide position 568 in a well-characterized white population. Polymorphisms were determined in 327 healthy, white individuals and 74 patients with sarcoidosis, using a PCR-sequence-specific primer assay. The serum KL-6 levels were measured by ELISA. Significant differences between serum KL-6 levels of healthy subjects who were grouped according to MUC1 568 genotype were observed (P<0.0001) (mean+/-SEM): AA (195.2+/-9.9 U/ml; 95% confidence interval [CI], 175.7-214.8), AG (246.0+/-8.6 U/ml; 95% CI, 229.0-263.1), and GG (302.6+/-11.8 U/ml; 95%CI, 279.3-326.0). In the patients with sarcoidosis, the results were (mean+/-SD): AA (550.1+/-411.7; 95% CI, 380.2-720.1), AG (716.3+/-452.4; 95% CI, 547.4-885.2), GG (1,151.0+/-1122; 95% CI, 610.1-1692.0); P=0.02. Comparison of the KL-6 levels in which the 568 genotype was ignored rendered 6 out of 74 (7.5%) misclassifications of "elevated" versus "normal" KL-6 levels or vice versa. In conclusion, the MUC1 568 A to G polymorphism may be of interest for diagnostic purposes because our study delivered in vivo evidence that it contributes to interindividual variations in KL-6 levels. PMID- 16357368 TI - The art and practice of women's imaging: power and potential. PMID- 16357366 TI - Restoration of mucociliary transport in the fluid-depleted trachea by surface active instillates. AB - Severe impairment of mucociliary transport (MCT) is a hallmark of cystic fibrosis (CF) lung disease. Recent studies demonstrate that pharmacologic inhibition of anion and liquid secretion in pig tracheas models the MCT defect in CF through depletion of the periciliary fluid component of airway surface liquid. In the present study, the effectiveness of various aqueous instillates on rehydration of the airway surface and restoration of MCT was assessed in this model. Excised porcine tracheas were mounted in a chamber that permitted simultaneous measurement of MCT and adventitial exposure of the airways to Krebs solution. When anion and liquid secretion were inhibited by treatment with bumetanide and dimethylamiloride, MCT was greatly reduced. Luminal instillation of aqueous solutions containing surface-active substances (1% Tween80 or calfactant) transiently restored MCT to high rates in nearly all tissues. Mucosal treatment with only Krebs solution or hypertonic saline restored MCT in only one half of the tracheas. We conclude that aqueous salt solutions alone can hydrate airway surfaces and restore MCT in some tissues, but surface-active substances may provide additional benefit in restoring MCT in this model of mucociliary stasis. We speculate that administration of surface-active substances, by aerosol or lavage, might help to restore MCT in the airways of patients with CF. PMID- 16357369 TI - Top 100 cited AJR articles at the AJR's Centennial. PMID- 16357370 TI - Inpatient radiology utilization: trends over the past decade. AB - OBJECTIVE: The objective of our study was to assess patterns of use of radiology services for inpatients at our institution between 1993 and 2002. MATERIALS AND METHODS: We retrospectively reviewed the administrative data about adult inpatients for fiscal years 1993-2002 in a 721-bed tertiary care institution. Examinations were coded according to imaging technique: conventional (plain films and fluoroscopy), sonography, nuclear medicine, CT, or MRI. We assessed workload trends using relative value units (RVUs). Linear regression analysis was used to assess the significance of trends for the number of examinations and RVUs per case-mix-adjusted admission (CMAA). RESULTS: There was a significant decrease in the total number of examinations per CMAA (p < 0.001). This was due to significant decreases in the use of conventional studies (p < 0.001) and sonograms (p < 0.001), despite significant increases in the number of nuclear medicine (p = 0.046), CT (p < 0.001), and MRI (p < 0.002) examinations per CMAA. The RVUs per CMAA increased significantly (p < 0.01) during the study period. CONCLUSION: Newer imaging technologies (nuclear medicine, CT, and MRI) are replacing older ones in the evaluation of inpatients. Despite the significant decrease in the total number of imaging examinations per CMAA, we observed a significant increase in RVUs per CMAA during the 10-year study period. Understanding the impact of this change in practice on the quality of care would be useful in justifying the increasing use of these new technologies, and decreasing their inappropriate use should be a priority in efforts that focus on controlling imaging expenditures. PMID- 16357371 TI - A portrait of pediatric radiologists in the United States. AB - OBJECTIVE: In recognition of the importance of pediatric radiology and the apparent shortage of radiologists in the field, the purpose of this study was to provide an extensive and detailed portrait of pediatric radiologists, their professional activities, and the practices in which they work. MATERIALS AND METHODS: We tabulated data from the American College of Radiology's 2003 Survey of Radiologists, a stratified random sample survey that achieved a 63% response rate with a total of 1,924 responses. Responses were weighted to make them representative of all radiologists in the United States. We compare information about pediatric radiologists with that for other radiologists. RESULTS: Approximately 3% of radiologists, some 800-900 physicians, are pediatric radiologists. Depending on how pediatric radiologist is defined, two thirds to three quarters of them spend 70% or more of their clinical work time doing pediatric radiology. Unlike other radiologists, a greater percentage of pediatric radiologists desire a reduction in workload (with a corresponding reduction in income) than desire an increase in workload. Pediatric radiologists who spend 70% or more of their clinical work time in their field are older than radiologists in general (average age, 55 vs 51 years), and the fraction of pediatric radiologists younger than 45 years is lower than for other subspecialists ( approximately 20% vs 37%). Pediatric radiologists are disproportionately women (one third or more, depending on definition, are women, vs 19% for other subspecialists and 15% for nonsubspecialists), hospital-based, in academic practices (approximately half vs one fifth for other subspecialists), and in the main cities of large metropolitan areas. CONCLUSION: A shortage of pediatric radiologists exists and is likely to intensify. Access to pediatric radiologists is probably a problem except for children in large metropolitan areas who connect readily to academic hospitals. Means to overcome these problems need to be actively sought. PMID- 16357372 TI - Hysterosalpingography and sonohysterography: lessons in technique. AB - OBJECTIVE: The objectives of this article are to review the examination techniques for hysterosalpingography and saline infusion sonohysterography and to present practical tips to enhance physician performance and minimize patient discomfort during these procedures. CONCLUSION: Optimally performed hysterosalpingography and saline infusion sonohysterography can be relatively comfortable procedures for the patient that provide a great deal of useful diagnostic information. PMID- 16357373 TI - High spectral and spatial resolution MRI of breast lesions: preliminary clinical experience. AB - OBJECTIVE: In previous research, high spectral and spatial resolution (HiSS) echo planar spectroscopic imaging (EPSI) was successfully applied to the human breast, obtaining improved contrast, anatomic detail, and sensitivity to contrast agents. To test HiSS in the clinical setting, we used HiSS MRI to image 30 women with suspicious breast lesions. SUBJECTS AND METHODS: Women with suspicious breast lesions were scanned before and after contrast administration using EPSI at 1.5 T (0.63-mm in-plane resolution, 2.6-Hz spectral resolution). Images with intensity proportional to the water signal peak height in each voxel were synthesized and compared with standard clinical fat-saturated and early dynamic subtraction images. Pre- and postcontrast HiSS images were compared to assess the effect of the contrast agent on water resonance structure. RESULTS: HiSS images scored significantly better than standard clinical images in lesion conspicuity, margin definition, and internal definition, even though they were acquired before contrast agent injection. Fat suppression was more complete and uniform and detail was shown on HiSS images more clearly than on conventional fat-saturation images. Thus, HiSS images often allowed easier evaluation of the lesion. Contrast agent-affected changes were often spatially and spectrally inhomogeneous. CONCLUSION: HiSS scans were successfully integrated into standard clinical examinations and provided diagnostically useful images before contrast agent injection. Thus, it might be possible to characterize suspicious lesions on the basis of precontrast high-resolution spectral information. This information and information about the effect of contrast agents could potentially improve the specificity of breast MRI. PMID- 16357374 TI - Image quality of a wet laser printer versus a paper printer for full-field digital mammograms. AB - OBJECTIVE: The purpose of our study was to compare the image quality of a wet laser printer with that of a paper printer for full-field digital mammography (FFDM). MATERIALS AND METHODS: For both a wet laser printer and a paper printer connected to an FFDM system, image quality parameters were evaluated using a standardized printer test image (luminance density, dynamic range). The detectability of standardized objects on a phantom was also evaluated. Furthermore, 640 mammograms of 80 patients with different breast tissue composition patterns were imaged with both printers. Subjective image quality parameters (brightness, contrast, and detection of details of anatomic structures that is, skin, subcutis, musculature, glandular tissue, and fat), the detectability of breast lesions (mass, calcifications), and the diagnostic performance according to the BI-RADS classification were evaluated. RESULTS: Both the luminance density and the dynamic range were superior for the wet laser printer. More standardized objects were visible on the phantom imaged with the wet laser printer than with the paper printer (13/16 vs 11/16). Each subjective image quality parameter of the mammograms from the wet laser printer was rated superior to those of the paper printer. Significantly more breast lesions were detected on the wet laser printer images than on the paper printer images (masses, 13 vs 10; calcifications, 65 vs 48; p < 0.05). With the paper printer images, BI-RADS 4 and 5 categories were underestimated for 10 (43.5%) of 23 patients. CONCLUSION: For FFDM, images obtained from a wet laser printer show superior objective and subjective image quality compared with a paper printer. As a consequence, the paper printer should not be used for FFDM. PMID- 16357375 TI - Intraductal papilloma: features on MR ductography using a microscopic coil. AB - OBJECTIVE: The purpose of this study was to describe the features of intraductal papilloma on MR ductography using a microscopic coil. CONCLUSION: Intraductal papilloma appeared in most cases as a well-circumscribed mass with early enhancement and delayed washout associated with a dilated duct, predominantly showing signal hyperintensity on T1- and T2-weighted imaging. PMID- 16357376 TI - Computer-aided detection as evidence in the courtroom: potential implications of an appellate court's ruling. AB - OBJECTIVE: The use of computer-aided detection (CAD) in radiology has been studied for different organ systems. As with any new technology, its impact on determinations of standards of clinical practice is an evolving one that is often defined by its acceptability not only in medical forums but also as defined by courts of law. CONCLUSION: We discuss the first known appellate legal decision regarding the acceptability of CAD as it relates to the clinical practice of mammography. PMID- 16357377 TI - An appellate court ruling and potential implications for CAD technology in the courtroom. PMID- 16357378 TI - Amyloidosis of the breast coexisting with ductal carcinoma in situ. PMID- 16357379 TI - Radiologic-pathologic conference of Brooke Army Medical Center: endometriosis of the canal of Nuck. PMID- 16357380 TI - The role of imaging during extracorporeal membrane oxygenation in pediatric respiratory failure. AB - OBJECTIVE: Extracorporeal membrane oxygenation (ECMO) is increasingly widely used in pediatric respiratory failure. Despite playing a key part in patient management during ECMO, the role of radiology is not widely reported. We discuss the principles of ECMO support and the normal imaging appearances. Radiologic findings arising from the complications of ECMO are highlighted. CONCLUSION: Radiology has a central role in establishing well-designed imaging protocols and vigilant reporting of ECMO apparatus positions and complications. PMID- 16357381 TI - Clinical and imaging mimickers of acute appendicitis in the pediatric population. AB - OBJECTIVE: The purpose of this article is to present the imaging appearance of common mimickers of appendicitis in children with right lower quadrant pain. CONCLUSION: The majority of children who undergo imaging for suspected appendicitis will end up having an alternative diagnosis. These mimickers can be gastrointestinal, genitourinary, or pulmonary. Familiarity with these alternative diagnoses can aid in the challenging task of imaging right lower quadrant pain in the pediatric population. PMID- 16357382 TI - Three-dimensional rotational angiography of neurovascular lesions in pediatric patients. AB - OBJECTIVE: In this pictorial essay, we review the 3D rotational angiography (RA) studies of six pediatric patients; in these cases, the information obtained with 3D RA was uniquely beneficial in diagnosis and treatment planning. CONCLUSION: Three-dimensional RA is an excellent tool for the evaluation of a number of intracranial lesions in pediatric patients: There is less total radiation exposure from a single rotational run than from CT or a conventional angiography examination that involves more than one view and the study is quick, with data acquisition requiring less than 8 sec and fully rendered 3D reconstructions generated within 180 sec. PMID- 16357383 TI - Assessment of two 3D MDCT colonography protocols for observation of colorectal polyps. AB - OBJECTIVE: The objective of our study was to assess the value of two-way interpretation (i.e., from rectum to cecum and vice versa) compared with one-way interpretation (i.e., from rectum to cecum only) in terms of polyp detection and interpretation time on MDCT colonography. MATERIALS AND METHODS: Fifty consecutive patients underwent both CT colonography and conventional colonoscopy. Three radiologists independently analyzed the CT colonographic examinations of each patient using a primary 3D method. All examinations were analyzed using two techniques: navigation from rectum to cecum only (one-way) and navigation from rectum to cecum and vice versa (two-way). Sensitivity and positive predictive value were calculated on both a per-polyp basis and a per-patient basis. Alternative free-response receiver operating characteristic (ROC) curve analysis was estimated, and image interpretation time was documented. RESULTS: One hundred fifty-five polyps were depicted in 45 patients by colonoscopy. The mean sensitivity of CT colonography for polyp detection with two-way (88.4%) was significantly superior to that with one-way (78.1%) (p < 0.01). The mean positive predictive value of each observer with one-way was 66.7%, whereas that with two way was 65.8%. The mean area under the alternative free-response ROC curve (A(z) value) with two-way (0.827) was higher than that with one-way (0.816), but there was not a statistically significant difference. The average interpretation time of each observer with two-way (39 min) was statistically significantly longer than that with one-way (25 min) (p < 0.01). CONCLUSION: When using a primary 3D interpretation technique at CT colonography, complete 3D navigation from rectum to cecum and from cecum to rectum is mandatory to maximize polyp detection. The image interpretation time for two-way interpretation is statistically significantly longer than that with one-way interpretation. PMID- 16357384 TI - CT colonography using 360-degree virtual dissection: a feasibility study. AB - OBJECTIVE: Using a 3D rendering technique called "virtual dissection," we sought to evaluate polyp and fold distortion using a colon phantom, estimate the polyp detection performance in humans, and estimate the added benefit of double interpretation and computer-aided diagnosis. MATERIALS AND METHODS: A colon phantom containing 144 polyps of varying sizes (5-12 mm) and shapes (flat, sessile, pedunculated) was scanned. Polyp shape and distortion at virtual dissection were categorized as flame, club, pea, or bizarre. Haustral fold distortion was graded. The CT colonography examinations in 20 consecutive patients (colonoscopically proven normal findings, n = 5; polyps > or = 1 cm, n = 17 in 15 patients) were blindly reviewed by three radiologists using the virtual dissection technique. The added benefits of double interpretation and computer aided diagnosis were tabulated. RESULTS: Sessile polyps appeared flame (35/48 [73%]) or pea (11/48 [23%]) in shape. Flat polyps appeared flame-shaped (31/47 [66%]) or pea-shaped (16/47 [34%]). Pedunculated polyps were flame (15/45 [33%]), club (20/45 [44%]), or pea (6/45 [13%]) in shape. Axial distortion occurred along the longitudinal axis. The sensitivities of the three observers for polyps of 1 cm or more were 16/17 (94%), 14/17 (82%), and 15/17 (88%). The specificities were 5/5 (100%), 5/5 (100%), and 4/5 (80%). Sensitivities after double interpretation and computer-aided diagnosis improved but did not reach statistical significance. CONCLUSION: Although distortion of colonic structures exists at virtual dissection, it does so in recognizable patterns, so that sensitivity for polyp detection is not compromised. PMID- 16357385 TI - Automated insufflation of carbon dioxide for MDCT colonography: distension and patient experience compared with manual insufflation. AB - OBJECTIVE: The purpose of our study was to compare the effects of automated and manual carbon dioxide insufflation before CT colonography on distention and patient acceptance. SUBJECTS AND METHODS: One hundred forty-one symptomatic subjects underwent CT colonography using either an automated device (n = 47) or a manual method (n = 94) for carbon dioxide insufflation. CT data sets were assessed retrospectively in consensus by two blinded observers who graded distention for six colonic segments using a 4-point scale. An additional assessment of the overall clinical adequacy of distention (yes/no) was also made, and any learning curve was sought. Each patient completed a validated 24-point patient questionnaire reflecting patient satisfaction and discomfort. Distention scores, clinical adequacy, and questionnaire responses were analyzed using ordered logistic regression, Fisher's exact test, and the Mann-Whitney test statistic, respectively. RESULTS: Automated insufflation significantly improved distention overall (p = 0.001). For individual segments, distention was significantly improved in the sigmoid (p = 0.007) and descending (p < 0.001) colons when the patient was supine; and in the sigmoid (p = 0.02), descending (p = 0.001), and transverse (p = 0.02) colons when supine and prone positions were combined. No significant difference was seen in the clinical adequacy of distention, nor was there evidence of any learning curve for either insufflation method. Subjects were more weary after automated insufflation (p = 0.03), but no significant difference was seen for the remaining 23 questionnaire items or for feelings of bloating or discomfort. CONCLUSION: Automated carbon dioxide insufflation significantly improves colonic distention compared with manual insufflation. Benefit is greatest in the left colon, particularly when the patient is supine. Patient acceptance is similar to that for manual insufflation. PMID- 16357386 TI - CT of acute biliopancreatic limb obstruction. AB - OBJECTIVE: Our objective was to report the CT features of biliopancreatic limb (afferent loop) obstruction. CONCLUSION: Acute biliopancreatic limb obstruction has typical CT features. Given its high morbidity and rate of reoperation, it is useful to make this specific diagnosis instead of reporting the findings as postoperative small bowel obstruction. PMID- 16357387 TI - CT findings for postsurgical blind pouch of small bowel. AB - OBJECTIVE: Our objective was to define the CT criteria for blind pouches formed after enteric anastomosis. CONCLUSION: Familiarity with the CT appearance of blind pouches avoids the mistaking of these entities for bowel obstruction or abscesses. PMID- 16357388 TI - Perfusion measurement in acute pancreatitis using dynamic perfusion MDCT. AB - OBJECTIVE: Our objective was to determine whether MDCT with perfusion imaging could help in assessing the severity of acute pancreatitis in the initial phase of the disease. One hundred six patients with abdominal pain were prospectively enrolled in this study. CONCLUSION: Patients were separated into two groups: P1 (severe) and P2 (mild) acute pancreatitis. Mean perfusion value was 24.8 mL/100 mL/min in the P1 group and 50.5 mL/100 mL/min in the P2 group (p = 0.0016, significant). Our preliminary data suggest that pancreatic perfusion measurement using MDCT with perfusion imaging could help in assessing the severity of acute pancreatitis. PMID- 16357389 TI - Colonic perforation at CT colonography in a patient without known colonic disease. PMID- 16357390 TI - Opacification of the genitourinary collecting system during MDCT urography with enhanced CT digital radiography: nonsaline versus saline bolus. AB - OBJECTIVE: The purpose of this study was to determine whether a saline bolus during CT urography improves urinary collecting system opacification and whether the addition of enhanced CT digital radiography (CTDR) improves urinary collecting system visualization with or without a saline bolus. MATERIALS AND METHODS: One hundred eight CT urography and enhanced CTDR examinations were reviewed. Fifty-four patients were given a saline bolus during CT urography, and 54 patients underwent CT urography without a saline bolus. Urinary collecting system opacification was evaluated by group (saline vs nonsaline), imaging technique (CT urography alone vs CT urography plus enhanced CTDR), number of enhanced CTDR images, and site of nonopacified urinary segments. Using a multivariate logistic regression model, we determined significance of variables and odds of complete opacification. RESULTS: In the saline group, 248 nonopacified sites were identified on CT urography alone and 95 sites with CT urography plus enhanced CTDR. In the nonsaline group, 185 nonopacified sites were identified on CT urography alone and 59 sites with CT urography plus enhanced CTDR. Combining both groups, 433 nonopacified sites were identified with CT urography alone and 154 sites with CT urography plus enhanced CTDR. Multivariate logistic regression showed significance for group (p = 0.010), imaging method (p < 0.0001), number of enhanced CTDR images (p = 0.048), and site of segment opacification (p < 0.0001). The renal pelvis shows the greatest odds and the distal ureter the lowest odds for complete opacification by group or imaging method. CONCLUSION: The addition of a saline bolus offers no improvement, whereas the addition of enhanced CTDR offers significant improvement in collecting system opacification during CT urography. PMID- 16357391 TI - Value of chemical shift subtraction MRI in characterization of adrenal masses. AB - OBJECTIVE: The purpose of this study was to assess the advantages of the image subtraction technique in chemical shift MRI for the differentiation of adrenal adenomas from nonadenomas. SUBJECTS AND METHODS: Thirty-five patients with 42 adrenal masses (eight metastases and 34 nonfunctioning adenomas) underwent chemical shift MRI using a double-echo fast low-angle shot sequence. Subsequently, opposed-phase chemical shift MR images were subtracted from in phase images. The subtraction images were assessed quantitatively and qualitatively. For quantitative assessment, the signal intensity values of the adrenal masses were measured by one investigator with manually defined regions of interest. Qualitative assessment of the subtraction images was performed independently by two investigators, who reported their confidence in diagnosing adenomas versus nonadenomas based on signal intensity of the adrenal masses on subtraction images. RESULTS: The mean signal intensities were significantly different between adenomas and metastases on subtraction images (213 vs 18; p < 0.0001). There was no overlap in signal intensities between adenomas and metastatic tumors. The accuracy in distinguishing adenomas from metastatic tumors was 100% if the cutoff value of the signal intensity selected was 36-106. Quantitative results corresponding to 100% specificity were also observed, with similar sensitivity. No difference in interpretation between the two investigators occurred. CONCLUSION: Chemical shift subtraction MRI provides a high confidence level in distinguishing adrenal adenomas from adrenal metastases. The image subtraction technique also facilitates quantitative and qualitative evaluation of adrenal masses in chemical shift MRI. PMID- 16357392 TI - Opacification of the collecting system and ureters on excretory-phase CT using oral water as contrast medium. PMID- 16357393 TI - Intraoperative sonographically guided needle localization of nonpalpable testicular tumors. PMID- 16357394 TI - Ferumoxtran-10-enhanced MR lymphangiography: does contrast-enhanced imaging alone suffice for accurate lymph node characterization? AB - OBJECTIVE: Ferumoxtran-10 is a lymphotropic MR contrast agent that is currently under investigation. It has been shown to be effective in staging lymph nodes of patients with various primary malignancies. The current technique with ferumoxtran-10 involves imaging before and 24 hr after contrast administration. The purpose of this study was to evaluate the accuracy of ferumoxtran-10-enhanced images alone in characterizing lymph nodes for oncologic staging 24 hr after contrast enhancement. MATERIALS AND METHODS: Seventy-seven patients (58 men, 19 women) with proven primary cancer (bladder [n = 20], breast [n = 10], endometrial [n = 1], renal [n = 3], penile [n = 4], prostate [n = 31], rectal [n = 1], testicular [n = 5], and ureteral [n = 2]) who were scheduled for surgical lymph node dissection were enrolled in the study. In these patients, 169 lymph nodes (mean size, 11.2 mm) were evaluated on T2*-weighted gradient-refocused echo MRI at l.5 T both before and 24-36 hr after the IV administration of ferumoxtran-10 (2.6 mg Fe/kg). Two blinded reviewers with differing levels of interpreting experience separately performed qualitative image evaluation. A 6-point scale was used to characterize lymph nodes on contrast-enhanced images alone and on combined unenhanced and contrast-enhanced images. Receiver operating characteristic (ROC) analysis was performed separately for both reviewers. RESULTS: Of the 169 lymph nodes evaluated, 55 were benign and 114 malignant by histopathologic analysis. The results of the ROC analysis comparing contrast enhanced images ([A(z) = area under ROC curve] reviewer 1, A(z) = 0.92; reviewer 2, A(z) = 0.94) alone with combined unenhanced and contrast-enhanced images (reviewer 1, A(z) = 0.94; reviewer 2, A(z) = 0.93) showed a statistically significant difference (p = 0.01) for reviewer 1 but no difference for reviewer 2 (p = 0.88). Reviewer 2 was more experienced in interpreting ferumoxtran-10 enhanced images than reviewer 1. CONCLUSION: On ferumoxtran-10-enhanced MR lymphangiography, contrast-enhanced images alone may suffice for lymph node characterization. However, a certain level of interpretation experience may be required before contrast-enhanced images can be used alone. PMID- 16357395 TI - Comparison of gadobenate dimeglumine-enhanced dynamic MRI and 16-MDCT for the detection of hepatocellular carcinoma. AB - OBJECTIVE: The objective of our study was to compare the diagnostic performance of gadobenate dimeglumine-enhanced MRI with that of 16-MDCT for the detection of hepatocellular carcinoma using receiver operating characteristic (ROC) curve analysis. MATERIALS AND METHODS: Thirty-one patients with 53 hepatocellular carcinomas underwent gadobenate dimeglumine-enhanced dynamic MRI and multiphasic CT using 16-MDCT within a mean interval of 5 days (range, 3-9 days). The dynamic MRI examination was performed using 3D fat-saturated volumetric interpolated imaging and sensitivity encoding on a 1.5-T unit. Both dynamic MRI and multiphasic MDCT included dual arterial phase images. Three observers independently interpreted the CT and MR images in random order, separately, and without patient identifiers. The diagnostic accuracy of each technique was evaluated using the alternative-free response ROC method. The sensitivity and positive predictive values were also calculated. RESULTS: The sensitivities of gadobenate dimeglumine-enhanced MRI for all observers were significantly higher than those of MDCT for all the lesions and for lesions 1.0 cm or smaller (p < 0.05); however, for lesions larger than 1.0 cm, the sensitivities of the two imaging techniques were similar. The mean area under the ROC curve (A(z)) of gadobenate dimeglumine-enhanced MRI (0.87 +/- 0.03 [SD]) was higher than that of MDCT (0.83 +/- 0.04), but no significant difference was found between them (p = 0.31). The number of false-positive findings on dynamic MRI was slightly higher than on MDCT, but no significant difference in the positive predictive value between the two imaging techniques was detected (observer 1, p = 0.06; observer 2, p = 0.13; observer 3, p = 1.00). CONCLUSION: Gadobenate dimeglumine-enhanced MRI has a higher sensitivity for small hepatocellular carcinomas ( 75 kg). The patients were assigned to one of three groups according to the injection protocol. The injection rate was alternated to 3 or 4 mL/sec in group 1. The injection duration was 38 or 47 sec in groups 2 and 3, respectively. The aortic arrival time and the 100-H threshold time in each patient were measured. The mean values and the variations in the aortic arrival time and 100-H threshold time according to the injection protocols and the contrast media were compared. RESULTS: The mean variations (+/- SD) of aortic arrival times and 100-H thresholds in group 2 (aortic arrival time = 16.1 +/- 2.7 sec, 100-H threshold time = 19.6 +/- 2.9 sec) were smaller than in groups 1 (16.3 +/- 3.0 sec and 19.9 +/- 3.7 sec, respectively) and 3 (16.8 +/- 3.5 sec and 20.4 +/- 4.1 sec, respectively). However, the range of aortic arrival times and 100-H threshold times was more than 10 sec for all groups. The mean aortic arrival time and 100-H threshold time for all patients were 16.5 and 20.0 sec, respectively, and did not vary significantly with the injection protocol and concentration of contrast medium. CONCLUSION: The individual variations of the aortic arrival and 100-H threshold times can be reduced using a fixed-duration injection technique, but there are still substantial variations. Therefore, a bolus-tracking technique is recommended for optimal timing of arterial phase scanning. PMID- 16357401 TI - Different reconstruction intervals for exclusion of coronary artery calcifications by retrospectively gated MDCT. AB - OBJECTIVE: Exclusion of coronary artery calcifications has a high negative predictive value for the diagnosis of coronary artery disease. However, it is known that significant differences in calcium scoring can occur because of the ECG trigger interval. Thus, the aim of the study was to evaluate the influence of different reconstruction intervals on detection of any coronary calcium by using MDCT and retrospective cardiac gating. CONCLUSION: For a true exclusion of coronary artery calcifications, different reconstruction intervals have to be evaluated. PMID- 16357403 TI - Importance of pretest probability score and D-dimer assay before sonography for lower limb deep venous thrombosis. AB - OBJECTIVE: The purpose of our study was to develop and validate a clinical score (the Hamilton score) for the assessment of lower limb deep venous thrombosis (DVT) and to determine the usefulness of this score and a D-dimer assay before a complete lower limb sonographic examination. SUBJECTS AND METHODS: Five hundred forty-two consecutive ambulatory patients presenting to the emergency department were prospectively recruited, of whom 16 patients were excluded from the study. Eighteen history and examination variables were collected by the emergency department physicians. The Simplify D-dimer assay and a complete, single lower limb sonographic examination were performed in all patients. All patients with a negative sonographic examination for DVT were followed up for 3 months, and all those with a positive sonographic examination were given anticoagulation therapy. The Hamilton score was developed using the data from the first 214 patients and was prospectively validated in the next 312 patients. RESULTS: The most significant factors associated with a diagnosis of DVT were immobilization of the lower limb, active malignancy, and a strong clinical suspicion of DVT without other diagnostic possibilities by the emergency department physicians. Other factors were bed rest or recent surgery, male sex, calf circumference difference greater than 3 cm, and erythema. The Hamilton score was developed with the following weights: immobilization of the lower limb (2 points), active malignancy (2 points), strong clinical suspicion of DVT without other diagnostic possibilities by the emergency physicians (2 points), bed rest or recent surgery (1 point), male sex (1 point), calf circumference difference greater than 3 cm (1 point), and erythema (1 point). A score of 3 or greater indicates a likely probability for DVT, and a score of 2 or less represents an unlikely probability for DVT. Of the 103 patients with an unlikely probability Hamilton score and a negative D-dimer assay, only one patient had isolated calf DVT. A combined diagnostic strategy of unlikely-probability Hamilton score and a negative D-dimer would have a negative predictive value of 99% (95% confidence interval, 94.7 100%). CONCLUSION: An unlikely-probability Hamilton score and a negative Simplify D-dimer assay effectively exclude lower limb DVT, and a sonographic examination is unnecessary in this group of ambulatory emergency department patients. PMID- 16357402 TI - Thrombus detection in the left atrial appendage using contrast-enhanced MRI: a pilot study. AB - OBJECTIVE: Left atrial thrombi are an important cause for embolism-related morbidity and mortality. Transesophageal echocardiography (TEE), the clinical reference, is semiinvasive; thus, we aimed to assess the value of contrast enhanced cardiovascular MRI for the detection of thrombus in the left atrial appendage. CONCLUSION: The image quality was good for both 2D perfusion (grade 4 +/- 1) and 3D turbo fast low-angle shot (FLASH) (grade 4 +/- 1, n.s.). Compared with TEE, 2D perfusion, 3D turboFLASH, and the combination of both techniques yielded sensitivities of 47/35/44%, specificities of 50/67/67%, positive predictive values of 73/75/80%, and negative predictive values of 25/27/29%, respectively. The size of the thrombus was overestimated by 2D perfusion (66%) and by 3D turboFLASH (25%) and agreement for location and shape of thrombus was 50% and 75% for 2D perfusion and 75% and 50% for 3D turboFLASH, respectively. The TEE thrombus size was significantly larger in patients with true-positive diagnoses by 2D perfusion (148%) and by 3D turboFLASH (151%) when compared with patients with false-negative diagnoses (p < 0.05 for both). No such difference was found for image quality, time delay between TEE and MRI examination, and location and shape of thrombi. Contrast-enhanced MRI lacks diagnostic accuracy for the detection of thrombi in the left atrial appendage. Future technical improvements are essential to establish this technique as a noninvasive alternative to TEE. PMID- 16357404 TI - Acute pulmonary embolism: correlation of CT pulmonary artery obstruction index with blood gas values. AB - OBJECTIVE: The purpose of our study was to investigate the relation between the pulmonary artery obstruction index assessed with helical CT and impairment in blood gases in patients with acute pulmonary embolism. SUBJECTS AND METHODS: Helical CT pulmonary angiography was performed in 78 patients who were suspected of having acute pulmonary embolism and selected as being free of underlying cardiopulmonary disease. Findings consistent with acute pulmonary embolism were observed in 34 patients. The severity was assessed by the pulmonary artery obstruction index, defined as Sigma (n x d), where n is the number of segmental arteries occluded and d is the degree of obstruction. Spearman's rank correlation coefficients were used to assess the correlation between the index of arterial obstruction and arterial partial pressure of oxygen (Pao(2)); alveolar-arterial difference in partial pressure of oxygen (Pao(2)-Pao(2)); arterial partial pressure of carbon dioxide (Paco(2)); and arterial oxygen saturation (Sao(2)). The statistical difference of the arterial blood gas values between the two groups of patients (those with and those without pulmonary embolism) was evaluated using the Mann-Whitney U test. Blood gases were comparatively evaluated below and above different index values (from 40% to 70%) and different Paco(2) values (25, 30, and 35 mm Hg) as possible indexes of embolism severity using the same test. The level of significance was set at 95% (p = 0.05). RESULTS: The values of Pao(2), Sao(2), and Paco(2) were significantly lower (p = 0.024, p = 0.0062, and p = 0.000075, respectively) and the values of Pao(2)-Pao(2) were significantly higher (p = 0.0169) in the pulmonary embolism group than in the no pulmonary-embolism group. A significant correlation was observed between the obstruction index and Pao(2) (r = -0.33, p = 0.05), Paco(2) (r = -0.34, p = 0.05), Pao(2)-Pao(2) (r = 0.39, p = 0.02), and Sao(2) (r = -0.35, p = 0.04). Using cutoff values for the pulmonary artery obstruction index of 40%, 50%, 60%, and 70%, we observed that Paco(2) and Pao(2)-Pao(2) differed significantly between above and below the 40% (p = 0.018 and p = 0.03), 50% (p = 0.0087 and p = 0.029), and 60% (p = 0.005 and p = 0.003) cutoffs. Pao(2) differed significantly for the cutoff values of 60% (p = 0.03) and 70% (p = 0.004). The same was observed for Sao(2) at 60% (p = 0.05) and 70% (p = 0.03). Comparisons for Paco(2) showed that a value of 30 mm Hg significantly separates levels of the pulmonary artery obstruction index (p = 0.002), with 78% sensitivity and 82% specificity indicating a pulmonary artery obstruction index greater than 50%. CONCLUSION: In patients with acute pulmonary embolism but no other underlying cardiopulmonary disease, the severity of the pulmonary arterial tree obstruction assessed using the CT obstruction index is significantly correlated to the blood gas values. The strongest correlation was observed between the index and the Pao(2)-Pao(2). Furthermore, a Paco(2) value of 30 mm Hg or less is highly suggestive of an obstruction index of more than 50% of the arterial bed. PMID- 16357405 TI - Tracheal metastasis of lung cancer: CT findings in six patients. AB - OBJECTIVE: Our objective was to assess CT findings of tracheal metastasis of lung cancer. CONCLUSION: Tracheal metastasis of primary non-small cell lung cancer manifested as an endotracheal nodule or eccentric wall thickening of the trachea, showing contrast enhancement with a predilection for the upper trachea on CT. During postoperative evaluation of patients with surgically resected lung cancer, the possibility of tracheal metastasis of lung cancer should be suggested when an endotracheal nodule or eccentric wall thickening is present on CT. PMID- 16357406 TI - Thoracic actinomycosis: CT features with histopathologic correlation. AB - OBJECTIVE: Thoracic actinomycosis is a chronic suppurative pulmonary or endobronchial infection caused by Actinomyces israelii, a gram-positive anaerobic organism. We present the CT features of thoracic actinomycosis with histopathologic correlation. CONCLUSION: The typical CT feature of parenchymal actinomycosis is a chronic segmental air-space consolidation containing necrotic low-attenuation areas with frequent cavity formation. A broncholith can be secondarily infected with Actinomyces organisms, resulting in endobronchial actinomycosis. It usually manifests as a proximal endobronchial calcification associated with distal obstructive pneumonia. PMID- 16357407 TI - Trauma cases from Harborview Medical Center. Airway injury after acute chlorine exposure. PMID- 16357408 TI - Measuring thyroid gland volume: should we change the correction factor? AB - OBJECTIVE: In the assessment of thyroid volume with sonography (formula of an ellipsoid), a correction factor is used. Whereas previously 0.524 was used, the World Health Organization has recently changed (after the first review) this correction factor to 0.479. We compare volume measurement of the thyroid using different correction factors to automated volume measurement using MDCT, and we define an optimal correction factor in thyroid volume assessment. CONCLUSION: Acceptable correction factors are situated in the range of 0.494-0.554. We propose a correction factor of 0.529 when using the ellipsoid formula. PMID- 16357409 TI - Diagnostic performance of MR arthrography after rotator cuff repair. AB - OBJECTIVE: The objective of this study was to investigate the diagnostic performance of MR arthrography after rotator cuff repair. MATERIALS AND METHODS: MR arthrographic examinations of the shoulder performed after rotator cuff repair and before revision surgery were retrospectively analyzed in 48 patients (31 males, 17 females; mean age, 50.3 years; age range, 17-69 years). Full-thickness and partial-thickness defects of the supraspinatus, infraspinatus, and subscapularis tendons were diagnosed independently by two radiologists. Revision surgery served as the standard of reference. RESULTS: Observer 1 correctly recognized five of eight intact supraspinatus tendons, 10 of 19 partial-thickness defects, and 19 of 21 full-thickness defects. For observer 2, the numbers were three of eight, eight of 19, and 18 of 21. The corresponding numbers for the infraspinatus tendon for observer 1 were 28 of 31, 0 of three, and 14 of 14 tendons. For observer 2, they were 28 of 31, two of three, and 11 of 14. For the subscapularis tendon, observer 1 made the correct diagnosis in 18 of 31, five of six, and nine of 11 tendons. The results for observer 2 were 26 of 31, one of six, and 10 of 11 tendons. Interobserver agreement (weighted kappa) was 0.47 for the supraspinatus, 0.64 for the infraspinatus, and 0.20 for the subscapularis tendons, respectively. CONCLUSION: Postoperative full-thickness defects of the rotator cuff are reliably diagnosed with MR arthrography. The diagnostic performance for partial-thickness defects is only moderate. PMID- 16357410 TI - Coracohumeral interval imaging in subcoracoid impingement syndrome on MRI. AB - OBJECTIVE: The coracohumeral interval previously has been described as predictive of subcoracoid impingement on dynamic screening. The purpose of this study was to determine whether a coracohumeral interval acquired from routinely performed MRI can reliably diagnose subcoracoid impingement. MATERIALS AND METHODS: Preoperative MRI examinations of 19 patients (16 males, three females) with subsequent surgical confirmation of subcoracoid impingement were reviewed retrospectively and compared with MRI studies of 41 control subjects (22 males, 19 females). Axial and oblique sagittal coracohumeral interval measurements were taken. The morphology of the coracoid process and lesser tuberosity was assessed. Postanalysis application of the data to two smaller groups of patients was performed. The first group consisted of nine subjects (three males, six females) for whom subcoracoid impingement was diagnosed prospectively on the basis of abnormalities found by MRI. The second group consisted of seven patients (two males, five females) who were referred for MRI evaluation because of clinically suspected subcoracoid impingement. RESULTS: The average coracohumeral interval for females was 3 mm smaller than that for males. Using sex-adjusted data, we found a statistically significant difference between individuals with or without subcoracoid impingement in the axial coracohumeral interval (p = 0.01). This value, however, was poorly predictive (area under the receiver operating characteristic curve, 0.73). An 11.5-mm axial coracohumeral interval had 84% sensitivity but only 44% specificity. A 10.5-mm axial coracohumeral interval had 79% sensitivity and 59% specificity. The shoulder morphologic features assessed and intraarticular contrast use were not statistically significantly related to the coracohumeral interval. In postanalysis application of data, in the group of nine subjects without clinical diagnosis of subcoracoid impingement, all prospective MRI subcoracoid impingement diagnoses were falsely positive. However, if subcoracoid impingement was the referring diagnosis, prospective MRI evaluation more often was correct (n = 7 [three true-negatives, two true positives, two false-negatives]). CONCLUSION: A sex-adjusted coracohumeral interval of 10.5-11.5 mm, although statistically significantly related to subcoracoid impingement, is poorly predictive of this diagnosis when acquired via routinely performed MRI. Subcoracoid impingement is primarily a clinical diagnosis that may be supported, but not established, by this means. PMID- 16357411 TI - Aggressive fibromatosis: MRI features with pathologic correlation. AB - OBJECTIVE: We present the MRI features with pathologic correlation of aggressive fibromatosis, incorporating 203 cases over a 5-year period from the Royal Marsden Hospital Sarcoma Unit database. MATERIALS AND METHODS: Sixty patients had imaging available for retrospective review of which 29 had preoperative MRI and final histopathologic diagnosis of aggressive fibromatosis. RESULTS: The average age at diagnosis was 41.3 years with a female-to-male sex ratio of 1.2:1. Twenty lesions were extraabdominal; six, intraabdominal; and three, in the abdominal wall (classic desmoid). The average tumor size was 6.4 cm (range, 2.2-13.7 cm). Intraabdominal aggressive fibromatosis produced the largest tumors, averaging 9.5 cm. Most lesions were ovoid (52%) or infiltrative (34.5%) in outline with an irregular or lobulated contour (76%). The lesions crossed major fascial boundaries in 31% of cases overall and in 66% of patients referred for recurrent disease. On MRI, homogeneous isointensity or mild hyperintensity on T1-weighted images and heterogenous high signal on T2-weighted or STIR images were seen. All lesions enhanced after IV gadolinium, usually avidly. In contrast to previous reports, 38% of cases failed to show low signal on all pulse sequences and no abnormalities were seen in local bone structures. Histology showed sheets of bland spindle cells in dense collagen and did not vary with the MRI signal characteristics of the lesion. Patients referred for recurrent disease were most likely to have a recurrence after surgery. MRI and pathology findings did not predict recurrence. CONCLUSION: Accurate diagnosis and staging of aggressive fibromatosis by MRI have important treatment and prognostic implications. PMID- 16357412 TI - Role of MRI in prevention of metatarsal stress fractures in collegiate basketball players. AB - OBJECTIVE: Metatarsal stress fractures are common and represent debilitating and potentially season-ending injuries for basketball players. Bone marrow edema is readily visualized on MRI and can be a sign of stress changes. Twenty-six asymptomatic male National Collegiate Athletic Association basketball players were imaged before the 2003-2004 season and 14 players were reimaged after the conclusion of the season with a screening study of long- and short-axis fat suppressed T2-weighted images (TR/effective TE, 3,500/56) to identify bone marrow edema in the metatarsals. CONCLUSION: Six (12%) of 52 feet showed a signal indicating bone marrow edema in the metatarsals. MRI depicts bone marrow edema in the feet before a fracture becomes evident. Identification of this edema may reveal stress changes, allowing early treatment and prevention of debilitating stress fractures. PMID- 16357413 TI - Posterior subtalar facet coalition with calcaneal stress fracture. PMID- 16357414 TI - MR arthrography is not proven to be preferred baseline MRI examination. PMID- 16357415 TI - Performance-based assessment of radiology faculty. PMID- 16357416 TI - Value of the single-phase technique in MDCT assessment of pancreatic tumors. PMID- 16357418 TI - 18F-FDG uptake on PET/CT in transposed ovaries. PMID- 16357422 TI - Renal oncocytoma displaying intense activity on 18F-FDG PET. PMID- 16357423 TI - Myoepithelioma of soft tissue originating from the hand: 18F-FDG PET features. PMID- 16357424 TI - Effects of averaging over motion and the resulting systematic errors in radiation therapy. AB - The potential for systematic errors in radiotherapy of a breathing patient is considered using the statistical model of Bortfeld et al (2002 Phys. Med. Biol. 47 2203-20). It is shown that although averaging over 30 fractions does result in a narrow Gaussian distribution of errors, as predicted by the central limit theorem, the fact that one or a few samples of the breathing patient's motion distribution are used for treatment planning (in contrast to the many treatment fractions that are likely to be delivered) may result in a much larger error with a systematic component. The error distribution may be particularly large if a scan at breath-hold is used for planning. PMID- 16357425 TI - An unexpected artefact with low-contrast high-energy film. AB - Kodak recently introduced new packaging for its X-Omat V verification film, including a label on the exterior of the packet. Patient images taken using the film in an image intensification cassette showed artefacts which appeared to be related to the label. Investigation showed the effect was only observed in conditions of high-Z build-up or backscatter or when the film was used without additional backscatter. The label provides extra build up when in front of the film and an increase in optical density of up to 0.04 units. When the label is on the rear of the film it absorbs backscattered particles, causing a decrease in optical density. It is concluded that X-Omat V film packets with labels are unsuitable for use in imaging cassettes unless they are used in conditions of low Z build-up. Alternatively the film must be removed from the labelled envelope if it is to be used in high-Z build-up conditions. PMID- 16357426 TI - Estimating perfusion using microCT to locate microspheres. AB - The injection of microspheres into the blood stream has been a common method to measure the spatial distribution of blood flow (perfusion). A technique to conduct this kind of measurement in small animal organs is presented using silver coated microspheres with a diameter of 16 microm and high-resolution computed tomography (microCT) to detect individual microspheres. Phantom experiments demonstrate the detectability of individual spheres. The distribution of microspheres within a rat heart is given as an example. Using non-destructive, three-dimensional imaging for microsphere detection avoids the cumbersome dissection of the organ into samples or slices and their subsequent registration. The detection of individual spheres allows high-resolution measurements of perfusion and arbitrary definition of regions of interest. These, in turn, allow for accurate statistical analysis of perfusion such as relative dispersion curves. PMID- 16357428 TI - Characterization of the phase-contrast radiography edge-enhancement effect in a cabinet x-ray system. AB - The purpose of this study was to demonstrate that a commercially available cabinet x-ray system is capable of phase-contrast radiography (PC-R) and to evaluate the effect of different system parameters on the degree of edge enhancement. An acrylic plastic edge phantom was imaged at different tube potentials (25-60 kV) and in different geometries (variable object-to-detector distances, R(2), at a constant source-to-detector distance, R(1) + R(2)). In addition, the effect of noise on the perceived edge enhancement was studied as a function of exposure time. Our results show that a modest degree of phase contrast can be achieved in an unmodified cabinet x-ray system. In addition, the particular system evaluated allowed low-noise PC-R images to be obtained with short (6 s or less) exposures. These results suggest that with appropriate geometric choices PC-R is already available to a wide range of research scientists for use in both small-animal and human-specimen experiments. PMID- 16357427 TI - An RF phased array applicator designed for hyperthermia breast cancer treatments. AB - An RF phased array applicator has been constructed for hyperthermia treatments in the intact breast. This RF phased array consists of four antennas mounted on a Lexan water tank, and geometric focusing is employed so that each antenna points in the direction of the intended target. The operating frequency for this phased array is 140 MHz. The RF array has been characterized both by electric field measurements in a water tank and by electric field simulations using the finite element method. The finite-element simulations are performed with HFSS software, where the mesh defined for finite-element calculations includes the geometry of the tank enclosure and four end-loaded dipole antennas. The material properties of the water tank enclosure and the antennas are also included in each simulation. The results of the finite-element simulations are compared to the measured values for this configuration, and the results, which include the effects of amplitude shading and phase shifting, show that the electric field predicted by finite-element simulations is similar to the measured field. Simulations also show that the contributions from standing waves are significant, which is consistent with measurement results. Simulated electric field and bio heat transfer results are also computed within a simple 3D breast model. Temperature simulations show that, although peak temperatures are generated outside the simulated tumour target, this RF phased array applicator is an effective device for regional hyperthermia in the intact breast. PMID- 16357429 TI - Adaptive prediction of internal target motion using external marker motion: a technical study. AB - An adaptive prediction approach was developed to infer internal target position by external marker positions. First, a prediction model (or adaptive neural network) is developed to infer target position from its former positions. For both internal target and external marker motion, two networks with the same type are created. Next, a linear model is established to correlate the prediction errors of both neural networks. Based on this, the prediction error of an internal target position can be reconstructed by the linear combination of the prediction errors of the external markers. Finally, the next position of the internal target is estimated by the network and subsequently corrected by the reconstructed prediction error. In a similar way, future positions are inferred as their previous positions are predicted and corrected. This method was examined by clinical data. The results demonstrated that an improvement (10% on average) of correlation between predicted signal and real internal motion was achieved, in comparison with the correlation between external markers and internal target motion. Based on the clinical data (with correlation coefficient 0.75 on average) observed between external marker and internal target motions, a prediction error (23% on average) of internal target position was achieved. The preliminary results indicated that this method is helpful to improve the predictability of internal target motion with the additional information of external marker signals. A consistent correlation between external and internal signals is important for prediction accuracy. PMID- 16357430 TI - Tumour oxygen dynamics measured simultaneously by near-infrared spectroscopy and 19F magnetic resonance imaging in rats. AB - Simultaneous near-infrared spectroscopy (NIRS) and magnetic resonance imaging (MRI) were used to investigate the correlation between tumour vascular oxygenation and tissue oxygen tension dynamics in rat breast 13762NF tumours with respect to hyperoxic gas breathing. NIRS directly detected global variations in the oxygenated haemoglobin concentration (Delta[HbO(2)]) within tumours and oxygen tension (pO(2)) maps were achieved using (19)F MRI of the reporter molecule hexafluorobenzene. Multiple correlations were examined between rates and magnitudes of vascular (Delta[HbO(2)]) and tissue (pO(2)) responses. Significant correlations were found between response to oxygen and carbogen breathing using either modality. Comparison of results for the two methods showed a correlation between the vascular perfusion rate ratio and the mean pO(2) values (R(2) > 0.7). The initial rates of increase of Delta[HbO(2)] and the slope of dynamic pO(2) response, d(pO(2))/dt, of well-oxygenated voxels in response to hyperoxic challenge were also correlated. These results demonstrate the feasibility of simultaneous measurements using NIRS and MRI. As expected, the rate of pO(2) response to oxygen is primarily dependent upon the well perfused rather than poorly perfused vasculature. PMID- 16357431 TI - Rapid dual-tracer PTSM+ATSM PET imaging of tumour blood flow and hypoxia: a simulation study. AB - Blood flow and hypoxia are interrelated aspects of physiology that affect cancer treatment and response. Cu-PTSM and Cu-ATSM are related PET tracers for blood flow and hypoxia, and the ability to rapidly image both tracers in a single scan would bring several advantages over conventional single-tracer techniques. Using dynamic imaging with staggered injections, overlapping signals for multiple PET tracers may be recovered utilizing information from kinetics and radioactive decay. In this work, rapid dual-tracer PTSM+ATSM PET was simulated and tested as a function of injection delay, order and relative dose for several copper isotopes, and the results were compared relative to separate single-tracer data. Time-activity curves representing a broad range of tumour blood flow and hypoxia levels were simulated, and parallel dual-tracer compartment modelling was used to recover the signals for each tracer. The main results were tested further using a torso phantom simulation of PET tumour imaging. Using scans as short as 30 minutes, the dual-tracer method provided measures of blood flow and hypoxia similar to single-tracer imaging. The best performance was obtained by injecting PTSM first and using a somewhat higher dose for ATSM. Comparable results for different copper isotopes suggest that tracer kinetics with staggered injections play a more important role than radioactive decay in the signal separation process. Rapid PTSM+ATSM PET has excellent potential for characterizing both tumour blood flow and hypoxia in a single, fast scan, provided that technological hurdles related to algorithm development and routine use can be overcome. PMID- 16357432 TI - Analytical reconstruction of deconvolved Fourier rebinned PET sinograms. AB - Fully 3D PET data are often rebinned into 2D data sets in order to avoid computationally intensive fully 3D reconstruction. Then, conventional 2D reconstruction techniques are employed to obtain images from the rebinned data. In a common scenario, 2D filtered back projection (FBP) is applied to Fourier rebinned (FORE) data. This approach is suboptimal because FBP is based on an idealized mathematical model of the data and cannot account for the statistical structure of data and noise. FORE data contain some blur in all three dimensions in comparison to conventional 2D PET data. In this work, we propose methods for approximating this blur in the sinogram domain due to FORE through its point spread function (PSF). We also explore simple methods for deconvolving the rebinned data with this PSF to restore it to a more ideal state prior to FBP. Our results show that deconvolution of the approximate transaxial PSF yields no improvement. When low image noise levels are required for detection tasks, the deconvolution of the axial PSF does not provide adequate resolution or quantitative benefits to justify its application. When accurate quantitation is required and higher noise levels are acceptable, the deconvolution of the axial PSF leads to considerable gains (30%) in accuracy over conventional FORE+FBP at matched noise levels. PMID- 16357433 TI - Enhancing the performance of model-based elastography by incorporating additional a priori information in the modulus image reconstruction process. AB - Model-based elastography is fraught with problems owing to the ill-posed nature of the inverse elasticity problem. To overcome this limitation, we have recently developed a novel inversion scheme that incorporates a priori information concerning the mechanical properties of the underlying tissue structures, and the variance incurred during displacement estimation in the modulus image reconstruction process. The information was procured by employing standard strain imaging methodology, and introduced in the reconstruction process through the generalized Tikhonov approach. In this paper, we report the results of experiments conducted on gelatin phantoms to evaluate the performance of modulus elastograms computed with the generalized Tikhonov (GTK) estimation criterion relative to those computed by employing the un-weighted least-squares estimation criterion, the weighted least-squares estimation criterion and the standard Tikhonov method (i.e., the generalized Tikhonov method with no modulus prior). The results indicate that modulus elastograms computed with the generalized Tikhonov approach had superior elastographic contrast discrimination and contrast recovery. In addition, image reconstruction was more resilient to structural decorrelation noise when additional constraints were imposed on the reconstruction process through the GTK method. PMID- 16357434 TI - Effect of ambient humidity on light transmittance through skin phantoms during cryogen spray cooling. AB - Cryogen spray cooling (CSC) is a technique employed to reduce the risk of epidermal damage during dermatologic laser surgery. However, while CSC protects the epidermis from non-specific thermal damage, it might reduce the effective fluence reaching the target chromophore due to scattering of light by the spray droplets and subsequent water condensation/freezing on the skin surface. The objective of this work was to study the effect of ambient humidity (omega) on light transmittance during CSC. An integrating sphere was employed to measure the dynamics of light transmittance through a deformable agar phantom during CSC. The study included two representative CSC spurt patterns studied using four omega: 57, 40, 20 and 12%. Results show that during CSC, as omega increased, light transmittance decreased. For the highest humidity level (57%) studied, light transmittance reached a minimum of 55% approximately 30 ms after spurt termination. In a controlled environment with omega = 12%, light transmittance reached a minimum of 87% approximately 30 ms after spurt termination. The reduced light transmittance immediately after spurt termination was most likely because of scattering of light caused by condensation of water vapour due to aggressive cooling of ambient air in the wake of the cryogen spurt. PMID- 16357435 TI - Diffuse reflectance spectrophotometry with visible light: comparison of four different methods in a tissue phantom. AB - The purpose of the study was to compare algorithms of four methods (plus two modifications) for spectrophotometric haemoglobin saturation measurements. Comparison was made in tissue phantoms basically consisting of a phosphate buffer, Intralipid and blood, allowing samples to be taken for reference measurements. Three experimental series were made. In experiment A (eight phantoms) we used the Knoefel method and measured specific extinction coefficients with a reflection spectrophotometer. In experiment B (six phantoms) the fully oxygenated phantoms were gradually deoxygenated with baker's yeast, and simultaneous measurements were made with our spectrophotometer and with a reference oxymeter (ABL-605) in 3 min intervals. For each spectrophotometric measurement haemoglobin saturation was calculated with all algorithms and modifications, and compared with reference. In experiment C (11 phantoms) we evaluated the ability of a modification of the Knoefel method to measure haemoglobin concentration in absolute quantities using extinction coefficients from experiment A.Results. Experiment A: with the Knoefel method extinction coefficients (+/-SD) for oxyhaemoglobin at 553.04 and 573.75 nm were 1.117 (+/ 0.0396) ODmM(-1) and 1.680 (+/- 0.0815) ODmM(-1), respectively, and for deoxyhaemoglobin 1.205 (+/- 0.0514) ODmM(-1) and 0.953 (+/-0.0487) ODmM(-1), respectively. Experiment B: high correlation with the reference was found in all methods (r = 0.94-0.97). However, agreement varied from evidently wrong in method 3 and the original method 4 (e.g. saturation above 160%) to high agreement in method 2 as well as the modifications of methods 1 and 4, where oxygen dissociation curves were close to the reference method. Experiment C: with the modified Knoefel method the mean haemoglobin concentration difference from reference was 8.3% and the correlation was high (r = 0.91). We conclude that method 2 and the modifications of 1 and 4 were superior to the others, but depended on known values in the same or similar phantoms. The original method 1 was independent of results from the tissue phantoms, but agreement was slightly poorer. Method 3 and the original method 4 could not be recommended. The ability of the modified method 1 to measure haemoglobin concentration is promising, but needs further development. PMID- 16357436 TI - Extraction of foetal ECG by combination of singular value decomposition and neuro fuzzy inference system. AB - Extraction of foetal ECG that is embedded in the maternal ECG is a challenging problem. This paper presents a combined system to extract foetal ECG from maternal abdominal ECG. The system uses a combination of singular value decomposition (SVD) and a neuro-fuzzy inference system. The SVD is used to construct two reference signals, while the fuzzy system is used as an adaptive canceller. The algorithm is applied on synthetic as well as real data and the results are presented. In addition, the paper presents an example of using the same system as a noise removal tool. PMID- 16357437 TI - Passive vortex currents in magneto- and electrocardiography: comparison of magnetic and electric signal strengths. AB - Vortex currents may be of importance in the early diagnosis of myocardial infarction caused by an occlusion of a coronary artery. We investigated the influence of a passive vortex current distribution, modelled by different conductivities in a hollow cylinder, on the localization error and on the signal strength in both the magnetocardiogram and the electrocardiogram. The hollow cylinder was mounted in a realistically shaped physical torso phantom. A platinum dipole was inserted into the cylinder. The compartment boundaries were modelled with two special ionic exchange membranes. The conductivity ratio of the cylinder compartment to the torso compartment was varied from 0.25 to 100. We compared the simultaneously measured magnetic and electric signal strengths as a function of this conductivity ratio. We found that an increasing conductivity ratio causes only a slight increase (about 19%) of the magnetic signal strength but a strong decrease (about 81%) of the electric signal strength. Using a homogeneous torso model, the dipole localization errors were, depending on the conductivity ratio, up to 16 mm. In conclusion, passive vortex currents might partially explain the differences between magnetocardiographic and electrocardiographic recordings observed both experimentally and clinically. PMID- 16357438 TI - A normal tissue dose response model of dynamic repair processes. AB - A model is presented for serial, critical element complication mechanisms for irradiated volumes from length scales of a few millimetres up to the entire organ. The central element of the model is the description of radiation complication as the failure of a dynamic repair process. The nature of the repair process is seen as reestablishing the structural organization of the tissue, rather than mere replenishment of lost cells. The interactions between the cells, such as migration, involved in the repair process are assumed to have finite ranges, which limits the repair capacity and is the defining property of a finite sized reconstruction unit. Since the details of the repair processes are largely unknown, the development aims to make the most general assumptions about them. The model employs analogies and methods from thermodynamics and statistical physics. An explicit analytical form of the dose response of the reconstruction unit for total, partial and inhomogeneous irradiation is derived. The use of the model is demonstrated with data from animal spinal cord experiments and clinical data about heart, lung and rectum. The three-parameter model lends a new perspective to the equivalent uniform dose formalism and the established serial and parallel complication models. Its implications for dose optimization are discussed. PMID- 16357439 TI - Spectral analysis of blood perfusion in the free latissimus dorsi myocutaneous flap and in normal skin. AB - To find the properties in the oscillatory components of the cutaneous blood flow on the successful free flap, a wavelet transform was applied to the laser Doppler flowmetry (LDF) signals which were measured simultaneously on the surfaces of the free latissimus dorsi myocutaneous flap and on the adjacent intact skin of the healthy limb, of 18 patients. The frequency interval from 0.0095 to 1.6 Hz was examined and was divided into five subintervals (I: 0.0095-0.021 Hz; II: 0.021 0.052 Hz; III: 0.052-0.145 Hz; IV: 0.145-0.6 Hz and V: 0.6-1.6 Hz) corresponding to endothelial metabolic, neurogenic, myogenic, respiratory and cardiac origins. The average amplitude and total power in the frequency range 0.0095-1.6 Hz as well as within subintervals I, II, IV and V were significantly lower for signals measured on the free flap than those obtained in the healthy limb. However in interval III, they were significantly higher. The normalized spectral amplitude and power in the free flap were significantly lower in only two intervals, I and II, yet in interval III they were significantly higher; no statistical significance was observed in intervals IV and V. The distinctive finding made in this study, aside from the decrease of endothelial metabolic processes and sympathetic control, was the significant increase of myogenic activity in the free flap. It is hoped that this work will contribute towards knowledge on blood circulation in free flaps and make the monitoring by LDF more reliable. PMID- 16357440 TI - Dose distribution to spinal structures from intrathecally administered yttrium 90. AB - Previous treatment of cerebrospinal fluid (CSF) malignancies by intrathecal administration of (131)I-radiolabelled monoclonal antibodies has led to the assumption that more healthy tissue will be spared when a pure beta-emitter such as (90)Y replaces (131)I. The purpose of this study is to compare and quantitatively evaluate the dose distribution from (90)Y to the CSF space and its surrounding spinal structures to (131)I. A 3D digital phantom of a section of the T-spine was constructed from the visible human project series of images which included the spinal cord, central canal, subarachnoid space, pia mater, arachnoid, dura mater, vertebral bone marrow and intervertebral disc. Monte Carlo N-particle (MCNP4C) was used to model the (90)Y and (131)I radiation distribution. Images of the CSF compartment were convolved with the radiation distribution to determine the dose within the subarachnoid space and surrounding tissues. (90)Y appears to be a suitable radionuclide in the treatment of central nervous system (CNS) malignancies when attached to mAb's and the dose distribution would be confined largely within the vertebral foramen. This choice may offer favourable dose improvement to the subarachnoid and surface of spinal cord over (131)I in such an application. PMID- 16357441 TI - Defining the roles of beta-catenin and plakoglobin in cell-cell adhesion: isolation of beta-catenin/plakoglobin-deficient F9 cells. AB - F9 teratocarcinoma cells in which beta-catenin and/or plakoglobin genes are knocked-out were generated and investigated in an effort to define the role of beta-catenin and plakoglobin in cell adhesion. Loss of beta-catenin expression only did not affect cadherin-mediated cell adhesion activity. Loss of both beta catenin and plakoglobin expression, however, severely affected the strong cell adhesion activity of cadherin. In beta-catenin-deficient cells, the amount of plakoglobin associated with E-cadherin dramatically increased. In beta catenin/plakoglobin-deficient cells, the level of E-cadherin and alpha-catenin markedly decreased. In these cells, E-cadherin formed large aggregates in cytoplasm and membrane localization of alpha-catenin was barely detected. These data confirmed that beta-catenin or plakoglobin is required for alpha-catenin to form complex with E-cadherin. It was also demonstrated that plakoglobin can compensate for the absence of beta-catenin. Moreover it was suggested that beta catenin or plakoglobin is required not only for the cell adhesion activity but also for the stable expression and cell surface localization of E-cadherin. PMID- 16357442 TI - Soluble form of heparin-binding EGF-like growth factor contributes to retinoic acid-induced epidermal hyperplasia. AB - Heparin-binding EGF-like growth factor (HB-EGF), a member of the EGF-family, is thought to be important for keratinocyte functions. HB-EGF is first synthesized as a membrane-anchored form, and its soluble form is released by ectodomain shedding. Here we investigate the role of HB-EGF in epidermal hyperplasia induced by all-trans retinoic acid (tRA) treatment. HB-EGF is normally expressed in epidermis of normal adult mice at very low levels, but topical tRA treatment results in epidermal hyperplasia, concomitant with the strong induction of HB-EGF expression in the suprabasal layer. tRA-induced epidermal hyperplasia was reduced both in the keratinocyte-specific HB-EGF null mice (K5-HB(del/del)) and knock-in mice expressing the uncleavable mutant form of HB-EGF (HB(uc/uc)), as compared with wild-type HB-EGF knock-in mice (HB(lox/lox)). Among ErbB tyrosine kinase receptors, EGF receptor (EGFR) and ErbB2 were selectively activated by tRA treatment in skin from wild-type mice, while the activation of these ErbB receptors was significantly reduced in the skin of HB-EGF null mice. These results indicate that expression of HB-EGF and generation of its soluble form, followed by activation of EGFR and ErbB2, are pivotal processes in tRA-induced epidermal hyperplasia. PMID- 16357443 TI - A role for fission yeast Rab GTPase Ypt7p in sporulation. AB - Ypt7p, a fission yeast (Schizosaccharomyces pombe) homologue of Rab7 GTPase, mediates fusion of endosomes to vacuoles and homotypic vacuole fusion. Here, we report that Ypt7p plays important roles in sporulation. Most ypt7Delta asci produced less than four spores, which were apparently immature and germinated at low frequency. Furthermore, ypt7Delta cells were defective in development of the forespore membranes. Vacuoles in sporulating cells were found to undergo extensive homotypic vacuole fusion to form a few large compartments occupying the entire cytoplasm of asci. This extensive vacuole fusion depended on Ypt7p. PMID- 16357444 TI - Antineutrophil cytoplasmic antibody-associated glomerulonephritis in type II diabetes mellitus. PMID- 16357445 TI - Lymphoid neoplasms associated with mosquito bites. PMID- 16357446 TI - Isoniazid and food interactions: --fish, cheese, and wine. PMID- 16357447 TI - Clinical significance of anti-CCP antibodies in rheumatoid arthritis. AB - A number of novel autoantibodies have been recently described in rheumatoid arthritis (RA), and their clinical significance and possible pathogenic roles have been discussed. In particular, new autoantibodies to citrullinated proteins such as filaggrin and its circular form (cyclic citrullinated peptide: CCP) are especially noteworthy because of their high sensitivity and high specificity. There are many studies that anti-CCP antibodies may serve as a powerful serologic marker for early diagnosis of RA and prognostic prediction of joint destruction. Anti-citrullinated protein antibodies are locally produced in RA joints, and citrullinated proteins (most are fibrins) are localized in RA synovial tissue. This finding strongly suggests a possibility that local citrullination of intraarticular proteins might be the initial event leading to autoantibody production in RA. Genetic factors such as a gene polymorphism of the citrullinating enzyme, PADI, might be associated with the breakage of self tolerance and induction of autoimmunity against citrullinated proteins. PMID- 16357449 TI - An outbreak of histamine poisoning after ingestion of the ground saury paste in eight patients taking isoniazid in tuberculous ward. AB - OBJECTIVE: To determine the cause of outbreak mimicking food poisoning, we studied the toxic polyamine contents of the food and analyzed the clinical characteristics of the affected eight patients. PATIENTS AND METHODS: Eight cases of histamine poisoning which occurred in tuberculous patients after dinner in our hospital were analyzed by clinical and biochemical methods. We examined the contents of four representative toxic polyamines, histamine, putrescine, cadaverine and tyramine, of the each food of the dinner and their serum concentrations of the monoamine oxidase (MAO), one of histaminases, using radioimmunoassay. RESULTS: The allergy-like symptoms such as flushing, headache, palpitation, itching, wheezing, dyspnea and diarrhea appeared from 20 minutes to two hours after ingestion in those eight patients taking isoniazid (INH), although the other 378 inpatients had no symptom. The histamine content of the ground saury paste was increased to 32 mg/100 g of food, however, the toxic level of food poisoning is less than 50 mg/100 g of food. All eight patients were taking INH, and their serum concentrations of MAO were decreased. CONCLUSION: We concluded that this accident was the histamine fish poisoning. We speculated that those allergy-like symptoms were due to both the increased histamine in the food made with the saury under poor storage conditions and the patients' reduced histaminase activities due to INH. We should perceive possible adverse effects depending on the interactions between certain drugs and food. PMID- 16357448 TI - Increased plasma levels of thioredoxin in patients with glucose intolerance. AB - OBJECTIVE: The aim of the present study was to determine the effects of glucose intolerance on oxidative stress in patients with coronary artery disease (CAD). METHODS: The patients were divided into 3 groups, diabetes mellitus (DM), IGT or normal glucose tolerance (NGT) according to the criteria of the American Diabetes Association. PATIENTS: The present study consisted of 178 consecutive patients who underwent diagnostic coronary arteriography and a 75-g glucose tolerance test. RESULTS: The level of plasma thioredoxin, a marker of oxidative stress was measured in every patient during the fasting state. The levels of plasma thioredoxin were significantly higher in the DM and IGT groups than the NGT group. Furthermore, we found that there was a positive association between thioredoxin levels and glycosylated hemoglobin (sigma=0.225, p=0.018). In multivariate logistic regression analysis, glucose intolerance (DM or IGT) was only independently associated with the high levels of thioredoxin. The levels of plasma thioredoxin were significantly higher in the CAD group compared to the non CAD group. In multivariate logistic regression analysis, high levels of thioredoxin, male, age and hypertension were independently associated with the presence of CAD. CONCLUSION: Glucose intolerance was associated with the high levels of thioredoxin. High levels of thioredoxin were related to the presence of CAD. The measurement of thioredoxin as the marker of oxidative stress may be useful for monitoring the development of the cardiovascular diseases. PMID- 16357450 TI - Circulating levels of soluble Fas ligand in cachexic patients with COPD are higher than those in non-cachexic patients with COPD. AB - OBJECTIVE: Apoptosis may be involved in the pathophysiology of cachexia in patients with chronic obstructive pulmonary disease (COPD). The purpose of this study is to assess the potential role of the Fas-Fas ligand (FasL) system in cachexic patients with COPD. PATIENTS AND METHODS: We measured the circulating levels of soluble FasL (sFasL), with a newly developed, highly sensitive enzyme linked immunosorbent assay system in seventy patients with COPD and forty-seven control subjects. RESULTS: The levels of sFasL in the COPD patients were significantly lower than those in the control subjects (46+/-29 vs. 55+/-28 pg/ml; p<0.05), whereas the levels of soluble Fas (sFas) remained unchanged between the two groups. The significant correlation between the levels of sFasL and sFas, observed in the control subjects (r=0.304; p<0.05), was absent in the COPD patients. Cachexic COPD patients with a relatively lower BMI (BMI <20 kg/m(2), n=45) and %fat (%fat <20%, n=34), showed significantly increased levels of sFasL compared to non-cachexic COPD patients with a relatively higher BMI (BMI > or =20 kg/m(2), n=25) and %fat (%fat > or =20%, n=36) (BMI; 51+/-33 vs. 36+/-15 pg/ml; p<0.05. %fat; 55+/-33 vs. 37+/-21 pg/ml; p<0.01), due to the inverse relationships between the body composition measurements and the levels of sFasL observed exclusively in the patients (BMI; r=-0.307; p<0.05. %fat; r=-0.283; p<0.05). CONCLUSION: These results may suggest that the Fas-FasL system does not play a significant role in the potential triggers of enhanced apoptosis leading to skeletal muscle wasting and adipose tissue depletion in cachexic patients with COPD. PMID- 16357451 TI - Cyclosporin treatment in steroid-resistant and acutely exacerbated interstitial pneumonia. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy of cyclosporin A (CsA) in patients with interstitial pneumonia (IP). DESIGN: Retrospective comparative study. PATIENTS: We reviewed 33 patients (23 males and 10 females with a mean age of 62.5 years) with histologically-proven progressive IP who were treated with CsA. All patients had corticosteroid-resistant IP or developed acute exacerbation of IP in their courses. RESULTS: The underlying systemic diseases were: idiopathic interstitial pneumonias (IIPs) in 19 patients, and collagen vascular diseases (CVDs) in 14. The histopathological patterns and underlying diseases of IP were classified as usual interstitial pneumonia (UIP)/idiopathic pulmonary fibrosis (IPF) in 10 patients, cellular-nonspecific interstitial pneumonia (NSIP)/IIPs in 3, fibrotic-NSIP/IIPs in 5, organizing pneumonia (OP)/IIP in 1, UIP/CVDs in 4, cellular-NSIP/CVDs in 7, fibrotic-NSIP/CVDs in 2, and diffuse alveolar damage (DAD)/CVD in 1, respectively. They received a low dosage of CsA combined with corticosteroids. The prognoses after treatment with CsA were well correlated with histopathological patterns. Cellular-NSIP and OP showed better prognoses than fibrotic-NSIP, UIP or DAD. In addition, CVDs had better prognoses than IIPs, when compared on the basis of the same histopathological patterns. Furthermore, the prognoses in the CsA-treated group were significantly better than in those without CsA treatment in regard to acute exacerbation of UIP/IPF. CONCLUSIONS: CsA combined with corticosteroids may be an efficacious treatment for corticosteroid-resistant IP and for acute exacerbation of IPF. PMID- 16357452 TI - Ten years of experience with liver transplantation for familial amyloid polyneuropathy in Japan: outcomes of living donor liver transplantations. AB - OBJECTIVE: We summarize 10 years of experience with liver transplantation for FAP patients in Japan and review the current opinions regarding this treatment for FAP. METHODS AND PATIENTS: All basic report data on patients at the time of transplantation were registered with the Japanese Liver Transplantation Society (JLTS). Based on the JLST report data, more detailed information on FAP patients was requested from each center. RESULTS: Living donor liver transplantation (LDLT) for FAP patients was first performed in Japan in 1993. LDLT has since been performed in 41 FAP patients, including nine cases of temporary auxiliary partial orthotopic liver transplantation (APOLT). Orthotopic liver transplantation (OLT) from cadaveric donors for FAP patients began in 1999, but only one FAP patient has subsequently undergone this procedure. Of these total of 43 FAP patients, 36 are currently alive: the one-year survival rate of patients after transplantation was 93%, and the five-year survival rate of these cases was 77%. Preoperative clinical severity and the nutritional status of patients are correlated with their outcome after liver transplantation. Domino (sequential) liver transplantation has been carried out in 20 domino recipients with end-stage liver diseases. Of the 20 domino recipients, 12 are currently alive. CONCLUSION: For FAP patients, these outcomes after the operation were very similar to those of OLT from cadaveric donors reported in other countries. Therefore, we concluded that for the treatment of FAP, LDLT from a living donor is equally effective as OLT from a cadaveric donor. PMID- 16357453 TI - Risk factors for hospital-acquired bacteremia. AB - OBJECTIVE: Bacteremia is one of the most serious health problems associated with high morbidity and mortality. The aim of this study was to identify risk factors for bacteremia in daily medical care to facilitate rapid and accurate clinical decisions about treatment. PATIENTS AND METHODS: We studied 306 inpatients retrospectively. Age, peripheral neutrophil count, C-reactive protein (CRP), platelets, serum total cholesterol, total protein, albumin and cholinesterase were compared in patients with positive- and negative-blood cultures. The associations between blood culture positivity and glucose tolerance, bedridden state, presence of a central venous catheter (CVC) or urinary catheter were examined. On October 14, 2002, strategies for prevention of catheter-related infection were altered in our hospital. We studied the impact of these changes on the risk of bacteremia. RESULTS: Sixty-seven patients had positive and 239 had negative blood cultures. Age, neutrophil, platelets, total protein, albumin, and cholinesterase were significantly different between the culture-positive patients and the culture-negative patients. Multivariate analysis showed albumin and platelets as independent predictors. The bedridden state and catheter-inserted states (central venous or urinary) conferred significantly higher positive blood culture rates. Multivariate analysis showed using urinary catheters and indwelling femoral CVCs as independent risk factors. There was no significant difference in the blood culture-positive rate before and after the change in prevention strategies; before the change, 6 of 9 catheter-inserted blood culture positive cases yielded MRSA, while 4 of 12 cultures yielded Staphylococcus epidermidis after the change. CONCLUSION: Our study highlights the risk factors of bacteremia in vulnerable patients. PMID- 16357454 TI - Pathological complete response in advanced gastrointestinal stromal tumor after imatinib therapy. AB - Gastrointestinal stromal tumor (GIST) is a rare neoplasm exhibiting, in most cases, mutations of c-kit. Recently it has been demonstrated that a majority of GIST patients with c-kit mutations respond to therapy with imatinib, a c-kit tyrosine kinase inhibitor. Although the response rate in patients treated with imatinib in prospective clinical studies is above 50%, complete response is rare, and the data on the use of imatinib as neoaduvant therapy facilitating radical surgery is still scanty. Here, we report on a patient with metastatic gastric GIST who underwent surgery after 6 months of imatinib therapy. No tumor cells were detected on pathological examination of resection specimen. This case report indicates that a pathological complete response could be achieved with imatinib therapy in patients with GIST, but a wider experience and longer follow-up is necessary to appreciate the prognostic significance of pathological complete response in GIST. PMID- 16357455 TI - Acute pancreatitis occurring in gastric aberrant pancreas treated with surgery and proved by histological examination. AB - We describe a case of gastric aberrant pancreas with acute pancreatitis. Barium meal examination, endoscopic examination and computed tomography of a 32-year-old man with abdominal pain revealed a submucosal tumor, about 3.5 cm in diameter, at the angulus of his stomach. Endoscopic ultrasonography revealed a hypoechoic mass with anechoic capillary areas. His serum amylase level was high at 262 IU/l. Laparoscopy-assisted local resection was carried out. The resected tumor revealed pancreatic tissue with extensive neutrophil infiltration in the gastric wall and fat necrosis in the subserosa. There are few cases of histologically proven acute pancreatitis in gastric aberrant pancreatic tissue. PMID- 16357456 TI - Type 1 diabetes mellitus and drug-resistant epilepsy: presence of high titer of anti-glutamic acid decarboxylase autoantibodies in serum and cerebrospinal fluid. AB - A 55-year-old man who was diagnosed as having type 1 diabetes mellitus (DM) at the age of 50 years was started on insulin therapy. At 54 years old of age, he suddenly developed complex partial seizures, which frequently occurred despite intensive anti-epileptic drug therapy. Neurological examination on admission revealed hyporeflexia in bilateral upper and lower extremities without any muscle rigidity, painful spasm or cerebellar ataxia. Laboratory examination showed poor glycemic control with increased glycated hemoglobin levels. Positive anti thyroglobulin antibodies and anti-thyroid peroxidase (TPO) antibodies and slight elevation of TSH levels are consistent with subclinical hypothyroidism due to Hashimoto's thyroiditis. A high titer of anti-glutamic acid decarboxylase (GAD) antibodies was detected in the patient's serum and cerebrospinal fluid (CSF). Electroencephalography showed temporal spikes, consistent with complex partial seizure. This is a very rare case presenting with concomitant type 1 diabetes and drug-resistant epilepsy associated with high titers of circulating and CSF anti GAD antibodies. PMID- 16357457 TI - Development of calciphylaxis after long-term steroid and methotroxate use in a patient with rheumatoid arthritis. AB - Calciphylaxis may be considered a small vessel vasculopathy which is generaly associated with end-stage renal disease and hyperparathyroidism. The precise pathogenesis of the disease is not known. It needs sensitizers and challengers to occur. Steroids and immunosuppressive drugs including methotrexate are among those challenger agents. Calciphylaxis in collagen vascular diseases is rare. Only one case in rheumatoid arthritis was recently reported. Here we describe a case of calciphylaxis associated with active rheumatoid arthritis. This patient had active disease despite treatment of steroids and methotrexate for a long time. She died shortly after the diagnosis of calciphylaxis due to sepsis. PMID- 16357458 TI - Adult Kawasaki disease unrelated to epstein-barr virus and group A Streptococcus. AB - A 23-year-old Japanese woman with a fever and generalized skin eruptions was referred to our hospital in July 1999. At admission, her temperature was 38.9 degrees C, and she had fluctuating symptoms including erythema of the extremities, conjunctival hyperemia, strawberry tongue, and generalized skin eruptions, but lymphadenopathy was not verified. An initially elevated urine leukocyte count (more than 100 per high power field) later returned to normal range without antibiotic therapy. Adult Kawasaki disease was diagnosed on the basis of the above symptomology. Echocardiograph showed transient effusion in the pericardium. Using the Harada scoring system for treatment of Kawasaki disease, we gave the patient aspirin and did not administer intravenous immunoglobulin. The clinical course was uneventful, and on the day of discharge (day 22 after onset), the laboratory test results were nearly normal. Laboratory test results were negative for both Epstein-Barr virus and group A Streptococcus. PMID- 16357459 TI - Lupus vasculopathy combined with renal infarction: unusual manifestation of lupus nephritis. AB - A 30-year-old woman with a 10-year history of systemic lupus erythematosus was admitted to our hospital because of the onset of hypertension and renal dysfunction. Renal arteriogram revealed multiple renal infarctions, and cut-off or tapering-stenosis in the interlobular arteries. Renal biopsy showed concentric intimal thickening with narrowed lumen in some arterioles and deposition of IgG/IgM/complement 3 in the wall of arteriole without any active lesions or immune complex deposition in glomeruli. The present case indicates that this type of renal vascular lesion in lupus nephritis, lupus vasculopathy, may cause renal infarction and the loss of renal function without active glomerular lesions. PMID- 16357460 TI - A probable case of aspiration pneumonia caused by Candida glabrata in a non neutropenic patient with candidemia. AB - Primary Candida pneumonia is rare, and detailed reports of Candida glabrata pneumonia have not been described. A 71-year-old woman had been treated for heart failure and developed aspiration pneumonia, which was refractory to antibacterial treatment. Antifungal treatment against C. glabrata resulted in resolution of pneumonia and candidemia. We report a probable case of C. glabrata pneumonia. PMID- 16357461 TI - Primary paraganglioma of the head of pancreas: contribution of combinatorial image analyses to the diagnosis of disease. PMID- 16357462 TI - Successful treatment with tacrolimus in a case of refractory dermatomyositis. PMID- 16357463 TI - An autopsy case of Loffler's endocarditis presenting a marked reduction in both ventricular cavities. PMID- 16357464 TI - Aplasia cutis congenita and skull defects after exposure to methimazole in utero. PMID- 16357465 TI - An adult case of acute disseminated encephalomyelitis accompanied with measles infection. PMID- 16357466 TI - Thermal aspects of melanistic and striped morphs of the snake Elaphe quadrivirgata. AB - Temperature is a critical factor limiting various aspects of the biology of ectotherms. In addition to environmental factors, coloration and body size are two physical properties that influence ectotherms' body temperature (T(b)). I compared the influences of these properties on thermal aspects of the two morphs of the color-dimorphic snake (E. quadrivirgata) under experimental conditions. First, I fitted T(b) data during heating to the von Bertalanffy equation, but considered parameter values of the equilibrium temperature obtained to be biologically meaningless. Alternatively, I limited the data for comparison of the morphs to T(b) < or =35 degrees C, which was the T(b) at which snakes began to move vigorously in the experiment. The rate of T(b) increase was significantly greater in the melanistic morph than in the striped morph. Heating rate was negatively correlated with body size in both morphs. The interaction of body size and heating rate did not significantly differ between the two morphs. The possibility of linkage, due to thermal advantage, between small body size and the prevalence of melanism in the population studied is briefly discussed. Rapid increase of T(b) is biologically advantageous because snakes with such ability would be released from various time and environmental constraints associated with thermoregulation under particular environmental conditions. PMID- 16357467 TI - Variations in body size of the lancelet Branchiostoma belcheri at different depths in the Seto Inland Sea: effect of food supply on the growth rate. AB - Variations in body size of the suspension-feeding lancelet Branchiostoma belcheri were examined from April 2001 to December 2001 at different water depths of 10-80 m. The lancelets were abundantly collected (220 indiv./dredge) at water depth of 10 m. The maximum size, ranging from 36.8 to 50.4 mm BL, decreased with increasing water depth. However, the minimum size of 7.4-7.8 mm BL, which corresponds to settling size, did not different with water depth. From seasonal changes in the length-frequency histograms, five cohorts were observed at every water depth. The lancelets grew to 44 mm BL at water depths of 10-20 m and to 34 mm BL at water depth of 80 m with four years of life span. Developmental stages of the gonads at an age of two years showed that no individuals had undeveloped gonads at water depth of 10 m, but 82% of them did at water depth of 80 m. The analysis of the stepwise multiple regressions of monthly growth rate on environmental variables showed that chlorophyll a was the best explanatory variable and showed a significant positive correlation with the growth of 1-2 years age groups. These results suggested that variations in the body length were mainly fluctuated by phytoplankton supply. PMID- 16357468 TI - Effects of melatonin feeding on smoltification in masu salmon (Oncorhynchus masou). AB - Influences of photoperiod on plasma melatonin profiles and effects of melatonin administration on long-day-induced smoltification in masu salmon (Oncorhynchus masou) were investigated in order to reveal the roles of melatonin in the regulation of smoltification in salmonids. Under light-dark (LD) cycles, plasma melatonin levels exhibited daily variation, with higher values during the dark phase than during the light phase. The duration of nocturnal elevation under short photoperiod (LD 8:16) was longer than that under long photoperiod (LD 16:8). Melatonin feeding (0.01, 0.1 and 1 mg/kg body weight) elevated plasma levels of melatonin in a dose-dependent manner for at least 7 h but not for 24 h. When masu salmon reared under short photoperiod were exposed to long photoperiod (LD 16:8) and fed melatonin (1 mg/kg body weight) 7 hours before the onset of darkness, a significantly smaller proportion of smolts appeared in the melatonin fed group after 32 days than in the control group. However, after 59 days of the treatment, there was no difference in the proportion of smolts between the control and melatonin-treated groups. Thus, melatonin feeding mimicked the effects of short photoperiod, which delays but does not completely suppress smoltification. These results indicate that the day length is transduced into changes in the duration of nocturnal elevation in plasma melatonin levels, and that artificial modification of the plasma melatonin pattern possibly delays the physiological processes of smoltification induced by long-day photoperiodic treatment. PMID- 16357469 TI - Ontogenetic changes in the capacity of the coral Pocillopora damicornis to originate branches. AB - Most colonial corals vary intraspecifically in growth forms, and the diversity in branching morphology is especially striking. While the effects of environmental factors on growth forms have been studied, the genetic control of coral branching patterns has received little attention. The discovery of ontogenetic changes in the capacity to originate branching would set the stage for studies of how branch formation is genetically controlled. During experiments investigating contact reactions in the coral Pocillopora damicornis, we observed that young colonies derived from settled planulae and colonies regenerated from adult branch tips assumed different growth forms. Young colonies formed at least one branch from the central region of the colony, while colonies regenerated from adult branch tips (3-5 mm long) did not form branches during the 9-month observation period. This pattern was invariable, regardless of the types and outcomes of the contact experiments or the orientation of the branch tips. However, some fragments taken from 1- or 2-year-old colonies formed branches. This suggests that the rate of branch formation in P. damicornis colonies decreases with age. These findings will facilitate investigations of the mechanism of coral branch formation at the molecular level. PMID- 16357470 TI - Phylogenetic studies of the subgenus Nialoe (s. lat.) (Coleoptera, Carabidae, Genus Pterostichus), part 1: Homology of the component parts of male genitalia and higher phylogeny. AB - A phylogenetic hypothesis of the subgenus Nialoe (s. lat.) of genus Pterostichus is proposed based on a cladistic analysis of seventeen morphological characters. Through comparative studies of the sclerites on the endophallus of male genitalia, it became apparent that the arrangement of these sclerites reflects seven different origins. The results show that Nialoe (s. lat.) is monophyletic and composed of four clades, while some traditional taxa are para- or polyphyletic, and are defined only by symplesiomorphies. Pterostichus (Nialoe) mosaicus sp. nov., which is indispensable for tracing the homology of sclerites in this subgenus, is described. PMID- 16357471 TI - Phylogenetic studies of the subgenus Nialoe (s. lat.) (Coleoptera, Carabidae, Genus Pterostichus), part 2: The Asymmetricus species group. AB - Pterostichus (Nialoe) asymmetricus Bates and its allies are revised based on the membranous parts of their genitalia. Four new taxa, P. (N.) ovaliphallus sp. nov., P. (N.) fujimurai ibukiyamanus ssp. nov., P. (N.) shotaroi kiimontanus ssp. nov., and P. (N.) basilobatus sp. nov. are described. A cladistic analysis based on 31 morphological characters reveals that Daisenilaoe (s. str.) Nakane and Straneo is included as one clade within Nialoe (s. str.). The speciation and dispersal process of this species group are also discussed. PMID- 16357472 TI - Morphological discrimination of two genetic groups of a Japanese salamander, Hynobius naevius (Amphibia, Caudata). AB - Hynobius naevius, distributed on western Honshu, Shikoku, and Kyushu Islands of Japan, includes two genetically distinct groups (Groups A and B) that have never been delimited morphologically. Using specimens from the entire species range, we investigated the possibility of distinguishing these groups morphologically. Multivariate analyses of morphometric characters resulted in recognition of two groups that corresponded well to the two genetic groups. One (Group A) was characterized by larger body, compressed tail, shallower vomerine tooth series, bluish- or reddish-purple ground color, and pale-white lateral markings. In contrast, another (Group B) was characterized by smaller body, cylindrical tail, longer vomerine tooth series, reddish-brown ground color, and white lateral markings. Group A was composed of populations from the Chugoku District of Honshu and northern Kyushu, and could not be divided into subgroups, while Group B encompassed populations from the Chubu and Kinki Districts of Honshu, Shikoku and Kyushu, and was subdivided into three local subgroups that are geographically separated by marine straits. Morphometric differentiation in Group A is presumed to have been less affected by genetic factors than by other factors, such as ecological relationships with other, coexisting species. Differentiation in Group B is assumed to have been enhanced not only by genetic but also by climatological factors. PMID- 16357473 TI - Plasma and urine levels of electrolytes, urea and steroid hormones involved in osmoregulation of cetaceans. AB - Cetaceans are well adapted to their hyperosmotic environment by properly developed osmoregulatory ability. A question here is how they regulate water and mineral balances in marine habitats. In the present study, we determined blood and urine levels of various chemicals involved in osmoregulation, compared them with those in artiodactyls, and characterized the values in the whales. Blood and urine samples obtained from baleen whales of common minke (Balaenoptera acutorostrata), sei (B. borealis), and Bryde's whales (B. brydei), and toothed whales of sperm whales (Physeter macrocephalus) were analyzed for osmolality, major electrolytes, urea, steroid hormones and glucose. The urine osmolality and Na(+) concentrations in the cetaceans were much higher than those in the cattle. Furthermore, the cetaceans had 5 to 11-fold urea in plasma than the cattle, and 2 to 4-fold urea in urine. There were no significant difference in the plasma concentrations of corticosteroids between the cetaceans and the cattle. The present results indicate that the osmoregulatory parameters seem to be not affected by the reproductive stage and sex steroid hormones. The concentrations of urea in plasma and urine of the baleen whales were higher than those of the sperm whales, indicating a possibility that their osmoregulatory mechanisms may be correlated to their feeding habits. The present results suggest that cetaceans have unique osmoregulatory mechanisms by which they excrete strongly hypertonic urine to maintain fluid homeostasis in marine habitats. PMID- 16357474 TI - Environmental factors affecting black/white coloration of the silken girdle in the swallowtail butterfly, Atrophaneura alcinous (Lepidoptera: Papilionidae). AB - The silken girdles of pupae of the swallowtail butterfly Atrophaneura alcinous show black and white color diphenism. Field observations revealed that all pupae observed on non-food plants and the leaves and stems of the larval food plant Aristolochia debilis were classified as a silken girdle of a black type, while a large portion of pupae pupating on the twigs and trunks of cherry trees in close proximity to A. debilis were classified as a silken girdle of a black type. Additionally, all pupae observed on the surfaces of artificial objects in areas where there are no surrounding plants or trees were classified as a silken girdle of a white type. We demonstrated the effect of day length and the texture, light, plant odor and humidity of pupation sites on the coloration of the silken girdle in A. alcinous. Regardless of long-day or short-day day length conditions, light conditions of constant light or dark, or the presence of a plant odor of A. debilis as environmental cues, all larvae placed at over 80% relative humidity (R.H.) developed into pupae with a silken girdle of a black type. However, all larvae developed into pupae with a silken girdle of a white type when R.H. was below 75%. Furthermore, when pupae with a silken girdle of a white type were transferred to conditions of 90% R.H. within 24 hr of pupation, the white color of the silken girdle changed into a black type within 24 hr of the transfer. The present data suggest that the induction of a black coloration of the silken girdle in A. alcinous requires a R.H. of approximately 80% or more as an environmental factor. PMID- 16357475 TI - A new species group in the genus Dichaetophora Duda (Diptera: Drosophilidae) based on a phylogenetic analysis, with descriptions of four new species from China. AB - A new species group, the sinensis group, is established within the genus Dichaetophora, based on a phylogenetic analysis of 37 adult morphological characters from 26 drosophilid species, including ten known species of Dichaetophora; four newly described species (Di. abnormis, Di. hainanensis, Di. bicornis, and Di. sinensis spp. nov. from China); and some representatives of related genera (Drosophila, Hirtodrosophila, and Scaptomyza). PMID- 16357476 TI - On the validity of the Japanese spider genus Metimorpha (Araneae, Araneidae). AB - The monotypic Japanese spider genus Metimorpha Strand, 1906 is known only from the juvenile holotype of M. tullia Bosenberg and Strand, 1906. The genus is not a tetragnathid as currently catalogued, nor a nephilid as tentatively suggested in the original description. Metimorpha is transferred to Araneidae, but the name is proposed as a nomen dubium. PMID- 16357477 TI - A new species of the genus Attheyella (Copepoda: Harpacticoida: Canthocamptidae) from Gosu cave in Korea. AB - A new species, Attheyella namkungi sp. nov., is described from Gosu cave at Danyang, Chungcheongbuk-do, South Korea. This species bears a superficial resemblance to A. coreana Miura, 1969 in having a 2-segmented mandibular palp with only four smooth apical setae, and in the armature formulae of the antennary exopod and thoracic legs. However the new species is clearly distinguishable by the non-sexually dimorphic caudal rami, two long plumose setae and a short seta on leg 6, and the absence of the setular row at the inner lateral margin of each caudal ramus. The new species is characterized from other congeners by having two adjacent inner spines on female leg 3 endopod-2, the caudal ramus twice as long as its width, absence of a barbed apophysis on male leg 3 endopod-2, and the seta number of legs 5 and 6. A key to the Korean species of the genus Attheyella is provided. PMID- 16357478 TI - Insuetifurca austronipponica, a new tardigrade (Eutardigrada: Macrobiotidae) from Kagoshima, southern Japan. AB - A new semiterrestrial tardigrade, Insuetifurca austronipponica, is described from Kakeroma Island, Kagoshima Prefecture, southern Japan. This is the third species known for the genus. It is distinguished from I. fujiensis primarily by a narrower buccopharyngeal tube and a longer pharyngeal tube, and from I. arrowsmithi primarily by a microplacoid clearly shorter than the macroplacoids. PMID- 16357479 TI - Plasma interleukin-18 levels in hemodialysis patients: increased by dialysis process and association with interleukin-6 and tumor necrotic factor-alpha. AB - BACKGROUND/AIMS: This study investigated changes of interleukin-18 (IL-18) levels before and after dialysis, and the possible association with other pro inflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha). METHODS: Plasma IL-18 of healthy controls, and pre- and post dialysis of uremic patients undergoing hemodialysis (HD) were evaluated by ELISA methodology. RESULTS: Plasma IL-18 levels were significantly increased in patients with maintenance HD (p <0.001) compared to its level in normal subjects. When compared to pre-dialysis levels, a significant increase in plasma IL-18 was measured at the end of HD (p = 0.032). There was a significant correlation among plasma IL-18, IL-6 and TNF-alpha levels in HD patients. CONCLUSION: Plasma IL-18 concentration was significantly higher in HD patients and was significantly elevated by cellulose-based HD processes. Pre-activation of immunologically active cells may contribute to the association between pre-dialysis IL-18 and post-dialysis IL-6 and TNF-alpha levels. PMID- 16357480 TI - Mycotic aneurysms of aortic root and aorta-to-left atrial fistula complicating bicuspid aortic valve endocarditis. AB - Unlike root abscess, fistula formation is quite uncommon in aortic valve endocarditis. In this report, we describe a patient with subacute bicuspid aortic valve endocarditis complicated by aortic insufficiency, mycotic aneurysms of the aortic root and fistulous communication between the aorta and the left atrium and his recovery upon surgical treatment. PMID- 16357481 TI - Association of the Phe206Leu allele of the L-selectin gene with coronary artery disease. AB - BACKGROUND AND AIMS: The aim of this study was to assess the association between the L-selectin Phe206Leu polymorphism and coronary artery disease. METHODS: A total of 322 patients (221 men and 101 women) with coronary artery disease in one or more vessels documented by angiography were studied; 157 subjects (85 men and 72 women) without atherosclerosis were included as controls. All subjects were genotyped for the L-selectin Phe206Leu gene polymorphism using polymerase chain reaction with sequence-specific primer (PCR-SSP). To assess disease severity, all patients were classified by numbers of coronary arteries with 50% stenosis. RESULTS: A significantly increased frequency of the 206Leu mutant allele was observed in patients with coronary artery disease compared to the controls. The 206Leu allele frequency occurred in 42% of the patients with coronary artery disease compared to 30% of the controls (p<0.009). No association was found between the severity of coronary artery disease and the L-selectin Phe206Leu polymorphism. CONCLUSION: Our findings suggest that carriage of L-selectin 206Leu mutant allele could contribute to susceptibility of Iranian individuals to contracting coronary artery disease. PMID- 16357482 TI - Twiddler's syndrome: an interesting cause of pacemaker failure. AB - This case report describes a patient with twiddler's syndrome, a rare condition of permanent pacemaker failure resulting from inadvertent or deliberate rotation of the pulse generator causing lead dislodgement. This report underscores the importance of dementia as a risk factor for twiddler's syndrome. PMID- 16357483 TI - Standard hemodiafiltration improves diuretic responsiveness in advanced congestive heart failure. PMID- 16357484 TI - Transient left ventricular apical ballooning. Report of 5 Israeli patients. AB - BACKGROUND: Transient left ventricular (LV) apical ballooning is characterized by acute onset of chest pain with reversible balloon-like LV motion abnormality, hypercontractile basal segments, ST segment elevation or T-wave inversion in anterior chest leads and mild cardiac enzyme rise in the absence of significant coronary disease. METHODS: We describe 5 patients (4 females) with anteroapical ballooning who were hospitalized with acute myocardial infarction and showed ST segment elevation in anterior chest leads. RESULTS: Echocardiogram demonstrated apical ballooning with normal or hypercontractile contraction of the basal segments. Four patients had severe mitral incompetence and one had mild incompetence. All patients had also systolic anterior motion and 4 had a significant LV outflow (LVOT) gradient. All patients underwent cardiac catheterization soon after admission showing non-significant narrowing of the coronary arteries. At discharge 4 patients had normal LV function and 1 was mildly impaired. CONCLUSIONS: LV apical ballooning is relatively rare. It should be suspected in older patients, mainly women, with severe mitral incompetence and LVOT gradient. PMID- 16357485 TI - 49,XXXXY syndrome with diabetes mellitus. AB - 49,XXXXY syndrome is a rare sex chromosome aneuploidy and characterized by mental retardation, skeletal defects, craniofacial anomalies and hypogonadism. The increased frequency of diabetes mellitus in patients with Klinefelter syndrome and other types of X-chromosome polysomy has been reported, but no cases of diabetes mellitus in adult with 49,XXXXY syndrome have been reported so far. We report an 18-year-old patient with 49,XXXXY syndrome accompanying diabetes mellitus. PMID- 16357486 TI - Factors determining patient choice of device for GH therapy. AB - AIM: To assess the factors determining patient choice of GH device, and whether offering free patient choice improves compliance with GH therapy. METHODS: A prospective cross-sectional study performed on patients offered free choice of GH device in a regional growth clinic. In a subgroup having home delivery, GH compliance was assessed using ampoule counts. RESULTS: 125 patients (74 (59%) male), median (range) 9.30 (1.0-18.3) years were commenced on GH from January 2001 to March 2004, and offered free choice of device. 68 (54%) chose a needled device, and 57 (46%) needle-free. There was no statistical difference in age, sex or diagnostic category between the two groups. Light blue devices were more likely to be chosen by males (p=0.056). Questionnaires giving reasons for choosing a device were available in 40, and a further 50 gave reasons for both choosing a specific device and not choosing others. Other than choice of needled/needle-free device, the factor most likely to determine choice was 'ease of use'. Only 6 (4.8%) subsequently changed device, and compliance remained high but unchanged at approximately 90%. CONCLUSIONS: There are no specific features which determine what GH device a patient will choose. For those units offering free patient choice, a wide range of different devices should be made available. PMID- 16357487 TI - Food intake of children with short stature born small for gestational age before and during a randomized GH trial. AB - Parents of short children born SGA often report that their children have a serious lack of appetite and a low food intake. In this study we investigated food intake, by using a standardized 7-day food questionnaire, in 88 short SGA children before start of GH treatment. The intake was compared with the recommended daily intake (RDI) of age-matched children. We also compared the food intake of GH-treated children (n=62) with randomized controls (n=26) after 1 year of GH treatment. In addition, we evaluated the effect of food intake and GH treatment on body composition and serum levels of IGF-I, IGFBP-3 and leptin. Our study shows that caloric intake, fat and carbohydrate intake of short SGA children aged 5.9 (1.6) years was significantly lower compared to the RDI for age matched children. One year of GH treatment resulted in a significant increase of caloric, fat, carbohydrate and protein intake compared to baseline. Compared to randomized controls, caloric, carbohydrate and protein intake increased significantly after 1 year of GH treatment. Short SGA children had significantly lower SDS scores for LBM, fat mass, skinfold (SF) and BMI compared to age-matched references. They also had significantly lower serum IGF-I, IGFBP-3 and leptin levels. GH treatment resulted in a significant increase of height, LBM, BMI, IGF I and IGFBP-3 SDS and a significant decrease of SF SDS and leptin SDS. In conclusion, our study shows that short SGA children have indeed a lower food intake than age-matched controls. During GH treatment the food intake increased significantly compared to baseline in contrast to the randomized control group. PMID- 16357488 TI - Prolactin suppresses GnRH but not TSH secretion. AB - BACKGROUND/AIMS: In animal models, prolactin increases tuberoinfundibular dopamine turnover, which has been demonstrated to suppress both hypothalamic GnRH and pituitary TSH secretion. To test the hypothesis that prolactin suppresses GnRH and TSH secretion in women, as preliminary evidence that a short-feedback dopamine loop also operates in the human, the effect of hyperprolactinemia on GnRH and TSH secretion was examined. METHODS: Subjects (n=6) underwent blood sampling every 10 min in the follicular phase of a control cycle and during a 12 hour recombinant human prolactin (r-hPRL) infusion preceded by 7 days of twice daily subcutaneous r-hPRL injections. LH and TSH pulse patterns and menstrual cycle parameters were measured. RESULTS: During the 7 days of r-hPRL administration, baseline prolactin increased from 16.0+/-3.0 to 101.6+/-11.6 microg/l, with a further increase to 253.7+/-27.7 microg/l during the 12-hour infusion. LH pulse frequency decreased (8.7+/-1.0 to 6.0+/-1.0 pulses/12 h; p<0.05) with r-hPRL administration, but there were no changes in LH pulse amplitude or mean LH levels. There were also no changes in TSH pulse frequency, mean or peak TSH. The decreased LH pulse frequency did not affect estradiol, inhibin A or B concentrations, or menstrual cycle length. CONCLUSION: These studies demonstrate that hyperprolactinemia suppresses pulsatile LH secretion but not TSH secretion and suggest that GnRH secretion is sensitive to hyperprolactinemia, but that TSH secretion is not. These data further suggest that the degree of GnRH disruption after 7 days of hyperprolactinemia is insufficient to disrupt menstrual cyclicity. PMID- 16357489 TI - Effect of Dracocephalum argunense on mast-cell-mediated hypersensitivity. AB - BACKGROUND: Stimulation of mast cells starts the process of degranulation resulting in release of mediators such as histamine and an array of inflammatory cytokines. In this report, we investigated the effect of an aqueous extract of Dracocephalum argunense Fisch. (Labiatae) (DAAE) on immediate-type hypersensitivity and studied its possible mechanisms of action, focusing on histamine release and proinflammatory cytokine expression in mast cells. METHODS: An in vivo model of systemic and local allergic reaction was investigated. Compound 48/80- or IgE-induced histamine release from mast cells was measured. The expression of proinflammatory cytokines such as TNF-alpha and IL-6 was measured by RT-PCR and ELISA. The level of intracellular calcium was measured by spectrofluorometry. NF-kappaB activation was measured by Western blot, EMSA and luciferase assay. RESULTS: DAAE inhibited systemic anaphylaxis, local allergic reactions, and serum histamine release in a dose-dependent manner in mice. DAAE dose-dependently reduced histamine release from mast cells activated by compound 48/80 or IgE. The inhibitory effect of DAAE on histamine release was mediated by the modulation of intracellular calcium. In addition, DAAE decreased TNF-alpha and IL-6 gene expression and production in human mast cells stimulated by phorbol 12-myristate-13-acetate (PMA) plus calcium ionophore A23187. The inhibitory effect of DAAE on the TNF-alpha and IL-6 expression was NF-kappaB dependent. CONCLUSION: Our findings provide evidence that DAAE inhibits mast-cell-derived immediate-type hypersensitivity. Taken together, the anti-allergic effect of DAAE in vivo and in vitro suggests a possible clinical use of this agent in immediate type hypersensitivity. PMID- 16357490 TI - Orange germin-like glycoprotein Cit s 1: an equivocal allergen. AB - BACKGROUND: Orange allergens are virtually unknown, in spite of the large consumption of this fruit. Germin-like proteins, together with vicilins and legumins, form the cupin superfamily of plant proteins, which includes many seed allergens. METHODS: Twenty-nine patients with allergy to oranges were studied. A major IgE-binding protein from orange extracts was isolated by means of a two step cation-exchange chromatographic protocol. The allergen was characterized by N-terminal amino acid sequencing and MALDI analysis, and its reactivity explored by specific IgE determination in individual sera, ELISA inhibition assays and in vivo skin prick tests (SPT). Chemical deglycosylation of the purified allergen was achieved by trifluoromethylsulfonate acid treatment. RESULTS: The 24-kDa purified allergen, designated Cit s 1, was identified as a germin-like glycoprotein, based on its N-terminal amino acid sequence, molecular size and recognition by rabbit anti-complex N-linked glycan antibodies. Specific IgE to Cit s 1 was detected in 62% of 29 individual sera from orange-allergic patients, whereas positive SPT responses to the purified allergen were obtained in only 10% of such patients. Deglycosylation of Cit s 1 resulted in a loss of its IgE binding capacity. Moreover, the unrelated plant glycoprotein horseradish peroxidase inhibited over 70% the IgE-binding to Cit s 1. CONCLUSIONS: Over 60% of patients with allergy to oranges show specific IgE to Cit s 1. However, the purified allergen exerts a low in vivo reactivity. Complex N-linked glycans seem to play a prominent role in the IgE-binding capacity of Cit s 1. This characteristic of Cit s 1, as well as of other orange glycoproteins, could lead to false positives if the diagnosis of allergy to oranges is mainly based on in vitro specific IgE determination. PMID- 16357491 TI - Application of newly developed amniotic membrane ointment for photorefractive keratectomy in rabbits. AB - We developed amniotic membrane ointment (AMO), and the effect of instilling the AMO after photorefractive keratectomy (PRK) was investigated with respect to inflammatory cell infiltration into the corneal stroma, apoptosis of keratocytes, and suppression of lipid peroxidation of cellular walls. The PRK procedure was performed on both eyes of 10 white rabbits. One eye of each rabbit (the experimental eye) was instilled with the AMO and the other eye of the rabbit (the control eye) with a base ointment 0, 8 and 16 h after the PRK procedure. Corneal specimens were collected 24 h after the PRK procedure. Hematoxylin-eosin stain and TUNEL assay were conducted to demonstrate polymorphonuclear and apoptotic cells, respectively. To assess lipid peroxidation, immunohistochemical staining with an antibody to malondialdehyde was undertaken. Compared to the control, the cornea instilled with the AMO had significantly less polymorphonuclear cells infiltrating into the corneal stroma as well as keratocytes subjected to apoptosis. These corneas also showed a significantly less extent of lipid peroxidation than the control. These data support that instillation of the AMO effectively reduced the recruitment of polymorphonuclear cells, the induction of apoptosis in keratocytes, and lipid peroxidation induced by PRK. Thus, this study could provide basic data on the clinical application of the AMO in the days ahead. PMID- 16357492 TI - Frequency of chromosome 17 aneuploidy in primary and recurrent pterygium by interphase-fluorescence in situ hybridization. AB - AIM: To investigate chromosome 17 numerical aberrations by using fluorescence in situ hybridization (FISH) in pterygia and to find out whether there is any association between chromosome 17 aneuploidy and recurrent pterygia. METHODS: Pterygium tissue samples were taken from 21 patients by surgical excision. Eighteen of them had primary and 3 had recurrent pterygium. Peripheral whole blood interphase cells obtained from 11 healthy subjects were assigned as control group. The cells from pterygium tissue and peripheral blood were incubated with a hypotonic solution and fixed in order to obtain interphase nuclei. FISH analysis with chromosome-17-specific alpha-satellite DNA probe was performed on both the interphase nuclei of pterygium tissue (of patients) and peripheral whole blood cells of controls. RESULTS: The mean percentage of chromosome 17 aneuploidy was 4.71% for the pterygia group and 4.41% for the controls. No significant difference of chromosome 17 aneuploidy was observed between the patients and the controls. When the group of patients with recurrences was compared with the group without recurrences, there was a significant difference in the frequency of chromosome 17 aneuploidy (U = 17, p = 0.029). CONCLUSIONS: Chromosome 17 aneuploidy is probably not an important factor in the formation of pterygium, but it may be related to recurrence. PMID- 16357493 TI - Conference report: Pittsburgh Pancreasfest 2005. PMID- 16357494 TI - Surgical treatment outcomes in subdural effusion: a clinical study. AB - OBJECTIVE: In this study, patients who underwent surgery due to subdural effusion were retrospectively analyzed. The location, depth and etiology of the subdural effusion, the surgical approach that was used and the recurrence rates were studied in these patients. METHOD: A total of 32 patients who were followed up and treated for subdural effusion at the Neurosurgery Clinic of the Yuzuncu Yil University School of Medicine were included in the study; 18 (56%) of the patients were male and 14 (44%) were female. The surgical techniques applied were surgical burr hole drainage, repeated subdural transaxial puncture and subduroperitoneal shunt approaches. The patients were evaluated by computerized tomography of the brain in week 1 and in the third month after surgery. Recurrences were evaluated based on radiological findings and the clinical condition of the patients. RESULT: The consciousness level of the patients was proportional to the mass effect of the subdural effusion. Lower recurrence rates were found in patients with a large midline shift resulting from the subdural effusion. In addition, recurrence rates were higher in patients with cerebral atrophy and lower protein content in the subdural effusion fluid. It was observed that these patients responded better to the subduroperitoneal shunt treatment. PMID- 16357495 TI - Prevalence of primary tethered cord syndrome associated with occult spinal dysraphism in primary school children in Turkey. AB - The prevalence and associated factors of primary tethered cord syndrome (PTCS) in primary school children were investigated. A cross-sectional study was performed in four demographically different primary schools in Turkey. Demographic, familial and physical data were collected from 5,499 children based on enuresis as a predominant symptom and dermatologic and orthopedic signs as clues of occult spinal dysraphism. Statistical analysis and input of the data were carried out with the SPSS package program 10.00, and logistic regression analysis was used to identify discriminating factors between enuretic children with or without neurologic signs. Of 5,499 analyzed children, 422 (7.7%) had enuresis nocturna, and 19.9% of 422 children had also daytime incontinence. Sixteen of these 422 enuretic children (3.8%) had several dermatologic signs. Five of them had spina bifida on plain radiographies, and 4 of them had cord tethering on lumbar MRI. Fifteen of 422 enuretic children (3.7%) had gait disturbances and orthopedic anomalies without cutaneous manifestations. Six of 15 children had spina bifida on plain graphies and 2 of them had tethered cord syndrome on MRI. The general prevalence of PTCS was found to be 0.1% of 5,499 analyzed children and 1.4% of enuretic children. A good outcome after untethering was found in 83.0% in this series. Practitioners should be aware of these clues of occult spinal dysraphism and resort to further radiologic and neurosurgical assessment. Early surgical intervention may halt the progression of the neurologic deficits and stabilize or reverse symptoms. PMID- 16357496 TI - The Chiari pseudotumor cerebri syndrome: symptom recurrence after decompressive surgery for Chiari malformation type I. AB - INTRODUCTION: The etiology of Chiari malformation type I (CM1) as well as other anomalies associated with CM1 remains poorly defined. We have noted the presence of elevated CSF pressures with small ventricles, consistent with the pseudotumor cerebri (PTC) syndrome in a group of CM1 patients that did not respond over the long term to posterior fossa decompression. In order to better understand this association, we reviewed a series of CM1 patients treated by posterior fossa decompression to define the prevalence and nature of post-Chiari PTC. METHODS: We performed a retrospective chart review of 192 patients diagnosed with CM1 and treated by posterior fossa decompression. Patients who failed to respond to surgery were evaluated by CINE MR flow studies to assess presence of CSF flow at the foramen magnum and then by lumbar puncture if flow was present. The diagnosis of Chiari PTC was defined by recurrence of Chiari-like symptoms after decompression, elevated lumbar CSF pressure in the absence of ventriculomegaly, and transient resolution of symptoms with large volume lumbar CSF drainage. RESULTS: Thirty-six of 192 patients did not improve with surgical decompression. Fifteen of 36 operative CM1 patients (41.6%) were found to have Chiari PTC. The most frequent symptoms of CM1/PTC patients were head pain, body aches, and balance difficulties. Three patients also experienced visual complaints. The mean maximum lumbar CSF pressure documented in this cohort was 26 cm of water in adults and 25.3 in children. All patients received treatment for the CM1/PTC that culminated with CSF shunt placement in 14/15. Seven of 9 pediatric patients had significant symptom resolution while 6/6 adult patients remained variably symptomatic. CONCLUSION: CM1 and PTC co-exist in a surprising percentage of failed operative CM1 patients and present with a syndrome that is difficult to treat. The etiology of this association after Chiari decompression is unclear, though perhaps posterior fossa surgery in the setting of abnormal anatomy and potentially anomalous CSF flow dynamics contributes to CSF malabsorption and resultant or coexistant PTC. PMID- 16357497 TI - Clinical and electroencephalographic features of infantile spasms associated with malformations of cortical development. AB - The aim of this study was to reveal the clinical and encephalographic (EEG) features of infantile spasms (IS) with malformations of cortical development (MCDs). The clinical features, EEG findings, neuroimaging studies and outcomes of various therapeutic modalities for 27 patients with IS and MCDs were reviewed. Background activities of EEG on the MCDs, i.e. asymmetric hypsarrhythmia, localized persistent polymorphic slowing, asymmetric slowing and diffuse fast activities, were shown in 22, 15, 9 and 2 patients, respectively. Partial epileptiform discharges such as localized paroxysmal fast activities, spindle shaped fast activities and subclinical seizures were shown in 15, 8 and 10 patients, respectively, and the lateralized prominence of generalized paroxysmal fast activities and generalized sharp and wave discharges in 4 and 5 patients, respectively. MCDs were suspected in 5 patients, as revealed by EEG and/or functional neuroimagings without distinct magnetic resonance imaging lesions, and confirmed by pathologic findings. Of the 11 patients treated with surgical resection, 8 became seizure free. EEG features can be very useful in the identification of underlying cortical dysgenesis in patients with IS. However, in one patient who underwent epilepsy surgery in early infancy, we observed easy bleeding and difficult hemostasis from friable vascular and parenchymal tissues. Various EEG features can be very useful in screening underlying MCDs. In addition, epilepsy surgery can be an effective therapeutic modality in many patients with otherwise medically intractable IS with MCDs. However, surgical intervention in extremely young infants should be performed with caution. PMID- 16357498 TI - Assessment of quality of continuous intracranial pressure recordings in children. AB - BACKGROUND: Bad quality of continuous intracranial pressure (ICP) recordings may potentially give wrong diagnostic information. METHOD: This study examined the ICP recordings of 33 children undergoing ICP monitoring during a 12-month period. Their ICP recordings were stored as raw data files and analyzed retrospectively. An automatic algorithm applied to every subsequent 6-second time window of each recording assessed the quality of ICP recordings. Time windows containing 4-18 cardiac beat-induced single ICP waves were accepted; time windows containing artifact-induced pressure waves due to noise in the signal were rejected. For every ICP recording, the number of accepted and rejected time windows were determined. In addition, the raw data ICP signal was assessed by visual inspection to verify the automatic algorithm. Two methods of computing mean pressure of a time window are compared; mean ICP was either computed according to current practice independent of the presence or absence of single ICP waves or computed only for single ICP waves within the time window (mean(SW) ICP). RESULTS: In 4 of the 33 patients (12%), more than 70% of the time windows of their ICP recordings were rejected; in 3 patients 95-100% of time windows were rejected. Visual inspection of the raw ICP signals confirmed the absence of single ICP waves. In these cases, computing mean(SW) ICP, not by mean ICP, revealed the bad signal quality. CONCLUSIONS: This retrospective study showed that in 3 of 33 patients (9%) the quality of the ICP recordings was so bad that they gave wrong diagnostic information. One way of obtaining quality control is to compute mean(SW) ICP, determined only for cardiac beat-induced single ICP waves within a time window. PMID- 16357499 TI - Spinal intradural-intramedullary cavernous malformation. Case report and literature review. AB - Cavernous angiomas or cavernomas are uncommon vascular malformations of the central nervous system and spinal involvement is much rarer especially in pediatric patients. We report a case of spinal intradural-intramedullary cavernous angioma in a 14-year-old male child. The cavernoma was located at the level of C6-C7 at the dorsal part of the spinal cord. The diagnosis was made with MRI and the patient underwent surgical treatment. The cavernoma was totally removed with laminotomy and microsurgical techniques. Somatosensory evoked potential monitoring was also used peroperatively. The clinical, radiological and surgical features of this rare case were presented and discussed with reference to the literature. PMID- 16357500 TI - Dual occurrence of pineal germinoma and testicular seminoma. Case report and review of the literature. AB - A 16-year-old male presented with obstructive hydrocephalus secondary to pineal germinoma. There have been many reported cases of abdominal metastasis of pineal germinoma after ventriculoperitoneal shunting. Endoscopic ventriculostomy was preferred in our case, thus avoiding iatrogenic peritoneal seeding, but spinal metastasis was unavoidable. Metastatic infiltration of the ventricular system and spinal meninges is the commonest mode of spread. Later, the patient underwent orchidectomy for an asymptomatic left testicular seminoma. He responded to chemotherapy, and had a complete recurrence-free remission for more than 10 years. As far as we know there are only a handful of reported cases of dual occurrence of two primary germ cell tumors (GCT), i.e. gonadal seminoma and pineal germinoma with spinal seeding. We also addressed the controversial subject of radiation versus chemotherapy in the management of patients with pineal germinomas. A review of the relevant literature and recommendations for future treatment of similar cases are discussed. PMID- 16357501 TI - Fatal haemorrhage in medulloblastoma following ventricular drainage. Case report and review of the literature. AB - Haemorrhage in medulloblastoma is reported to be very rare. The authors report a case of a 13-year-old boy who presented with headache, unsteadiness, diplopia and papilloedema due to posterior fossa medulloblastoma causing obstructive hydrocephalus. Six hours following placement of an external ventricular drain, he suddenly became comatose with respiratory arrest. The cause was marked upward herniation of the anterior vermis and downward herniation of the cerebellar tonsils due to massive spontaneous intratumoural haemorrhage extended into the ventricular system. This atypical clinical course of fatal haemorrhage in medulloblastoma after insertion of external ventricular drainage is reported and the literature discussed. PMID- 16357502 TI - Giant terminal lipomyelocystocele. AB - Terminal myelocystocele is a rare form of occult spinal dysraphism in which the hydromyelic caudal spinal cord and the subarachnoid space are herniated through a posterior spina bifida. A 1-year-old female child presented with a large lumbosacral mass (30 x 20 x 10 cm), flaccid paraplegia and urinary incontinence since birth. Magnetic resonance imaging revealed a low-lying conus (with associated conus lipoma) and a dilated central canal surrounded by a meningocele suggestive of terminal lipomyelocystocele and was operated on successfully. In our experience, this was a giant terminal lipomyelocystocele and such a large lesion has not been reported in the literature before. Terminal myelocystocele should be included in the differential diagnosis of congenital lesions presenting as a lumbosacral mass and operated early. PMID- 16357503 TI - Neurenteric cyst of the upper cervical spine: excision via posterior approach. AB - Neurenteric cyst of the cervical spine is an uncommon congenital lesion and usually associated with many craniocervical anomalies. Solitary neurenteric cyst of the upper cervical spine is very rare. Surgery is the main treatment method. An 11-year-old female child presented with the symptoms of neck pain that radiated to both shoulders and arms. Magnetic resonance imaging of the patient showed a cystic mass lesion ventral to the spinal cord and lying at the level from C1 to C3. She underwent surgical resection via a posterior approach and complete cyst resection was performed. Although removal of a neurenteric cyst through a posterior approach may result in spinal cord injury, this method is easier and safer than the anterior approach, especially in children. PMID- 16357504 TI - Alagille syndrome and aneurysmal subarachnoid hemorrhage. Case report and review of the literature. AB - The authors report the case of a 21-year-old female with a known history of Alagille syndrome (AGS) who was found to have a basilar terminus aneurysm without evidence of rupture. Prior to intervention, the patient's hospital course became complicated by multiple medical problems associated with AGS. Subsequently, the patient had an acute neurological decline. An unenhanced CT of the head demonstrated diffuse subarachnoid hemorrhage, intraparenchymal hematoma and intraventricular hemorrhage. AGS is an autosomal dominant arteriodysplastic syndrome with multiple organ system involvement caused by a mutation in the Jagged1 gene. Intracranial hemorrhage is one of the many complications observed in this patient population. While there are multiple case reports in the literature reviewing the spectrum of cerebrovascular events and abnormalities, intracranial aneurysmal rupture has only recently been described. To our knowledge, this is the third reported case of documented aneurysmal subarachnoid hemorrhage in a patient with AGS. The authors present a brief review of the vascular abnormalities both intracranial and systemic seen in AGS. The genomic abnormalities of this syndrome are also reviewed with particular attention to the Jagged1 gene and the Notch receptor signaling pathway which may reveal elements of the pathophysiology involved in aneurysm formation and rupture in AGS patients. In light of the increased incidence of intracranial hemorrhage in AGS and the possible link to aneurysmal subarachnoid hemorrhage, establishing the incidence of intracranial aneurysms in AGS and the role of screening these patients is indicated. PMID- 16357505 TI - Non-operative management of benign intracranial hypertension presenting with complete visual loss and deafness. AB - Benign intracranial hypertension (BIH) may lead to blindness and rarely deafness. We describe the case of a rapidly deteriorating 14-year-old African girl who presented with headaches associated with complete visual and hearing loss due to BIH. This was managed non-operatively with lumbar cerebrospinal fluid tap, weight reduction, nicotinic acid and acetazolamide. Response to treatment was quite dramatic with resolution of severe headaches and regaining of light perception 8 days after commencing treatment. By 3 months hearing recovered to normal and there was resolution of vision. This to the best of our knowledge is the first reported case of complete visual and hearing loss occurring in a patient with BIH, which was managed successfully non-operatively. When indicated, non operative management is an effective treatment option even in malignant BIH. PMID- 16357506 TI - Hemorrhage into the brain abscess cavity with Fallot's tetralogy. AB - Hemorrhage into the abscess cavity is a complication of brain abscess. It has been reported to be due to inflammation which results in the damage of the fragile neovasculature of the abscess wall. Hypoxia caused by Fallot's tetralogy or other congenital heart diseases facilitates the damage of these vessels with the lacking supportive tissues, and in turn intracavital bleeding. PMID- 16357508 TI - Tumor-targeted therapy with a conditionally replicating mutant of HSV-1 induces regression of xenografted human hepatomas. AB - The selectively oncolytic effects of mtHSV, a HSV icp34.5 mutant with lacz gene insertion, on several tumor cells in vitro and its antitumor effects by the intratumoral (IT) route to nude mice loaded the human hepatoma xenografts were explored. The mtHSV could conditionally replicate in and lyse Hep-3B (human hepatoma cells), Hep-2 (human larynx cancer cells) and SPC-A1 (human lung cancer cells), but not MRC-5 (human fibroblast cells). The 125 nude mice loaded with Hep 3B were randomly divided into five treatment groups and given three IT injections with three different dose of the mtHSV, adriamycin (ADM), or vehicle (supernatant of non-infected Vero cells). Significant tumor growth inhibition (30%-70%) was seen in the nude mice treated IT with mtHSV, whereas tumors treated IT with Vero supernatant displayed rapid tumor growth. The results of regular and biochemical blood examination, systemic necropsy and pathological slices showed that mtHSV almost has no side effect on treated mice. RT-PCR results revealed that the replication of mtHSV was exclusively confined to the treated tumors, but not to other organs. Our results provide further preclinical evidence that mtHSV may be used as an oncolytic agent for cancer therapy. PMID- 16357509 TI - KLF4 and KLF5 regulate proliferation, apoptosis and invasion in esophageal cancer cells. AB - KLF4 and KLF5, members of the KLF family of transcription factors, play key roles in proliferation, differentiation, and carcinogenesis in a number of gastrointestinal tissues. While KLF4 is expressed in differentiating epithelial cells, KLF5 is found in proliferating cells of the gastrointestinal tract, including the esophagus. KLF4 regulates a number of genes vital for esophageal epithelial differentiation, and decreased expression of KLF4 is seen in esophageal squamous cancers. Nonetheless, the roles of KLF4 and KLF5 in esophageal tumor progression are not known. Here, using TE2 cells stably infected with retroviral vectors to express KLF4 or KLF5, we demonstrate that KLF4 and KLF5 are key players in a number of cellular processes critical for esophageal carcinogenesis. TE2 cells, derived from a patient with poorly differentiated esophageal squamous cancer, normally lack KLF4 and KLF5. Expression of KLF5 in TE2 cells inhibits proliferation, and both KLF4 and KLF5 decrease viability after treatment with hydrogen peroxide and increase anoikis. In response to DNA damage from UV irradiation, viability is decreased in KLF5 but not KLF4 infected cells. Both KLF4 and KLF5 upregulate the cdk inhibitor p21(waf1/cip1) following UV irradiation, but the pro-apoptotic protein BAX is markedly induced only by KLF5. Thus KLF4 may preferentially activate DNA repair pathways while KLF5 induces both DNA repair and apoptosis after UV irradiation. Expression of KLF4 or KLF5 in TE2 cells also inhibits invasion, consistent with a role for each in preventing tumor metastasis. In summary, KLF4 and KLF5 regulate esophageal carcinogenesis by affecting proliferation, apoptosis, and invasion. PMID- 16357510 TI - KLF4 and PCNA identify stages of tumor initiation in a conditional model of cutaneous squamous epithelial neoplasia. AB - KLF4 is induced upon growth-arrest in vitro and during epithelial maturation in vivo, and is essential for proper cell fate specification of post-mitotic cells. In spite of a normal role in post-mitotic cells, expression is upregulated and constitutive in certain tumor types. KLF4 functions as an oncogene in vitro, and enforced expression in basal cells of mouse skin rapidly induces lesions similar to hyperplasia, dysplasia and squamous cell carcinoma (SCC). Here we used conditional expression to characterize early steps in KLF4-mediated tumor initiation. In contrast to SCC-like lesions that result when using a conditional, keratin 14 promoter-dependent strategy, lower conditional expression achieved using a MMTV promoter induced only epidermal cycling within morphologically normal skin, a process we termed occult cell turnover. Surprisingly, KLF4-induced hyperplastic lesions showed increased transgene-derived mRNA and protein in maturing, PCNA-negative cells, a property of endogenous KLF4. In contrast, hyperplastic lesions induced by GLI1, a control, showed uniform transgene expression. In KLF4-induced dysplasia and SCC the complementarity of KLF4 and PCNA was replaced by concordance of the two proteins. These studies show that KLF4 transcripts are normally suppressed in cycling cells in a promoter independent fashion, consistent with a post-transcriptional control, and reveal loss of this control in the transition from hyperplasia to dysplasia. Like the mouse tumors, human cutaneous SCCs and adjacent dysplasias frequently showed maturation-independence of KLF4, with co-expression of KLF4 and PCNA. A smaller subset of human SCCs showed complementarity of KLF4 and PCNA, similar to hyperplastic mouse skin. The results identify parallels between a mouse model and human primary tumors, and show that successive increases of KLF4 in the nuclei of basal keratinocytes leads to occult cell turnover followed by hyperplasia, dysplasia, and invasive SCC. PMID- 16357511 TI - Cooperation between BRCA1 and p53 in repair of cyclobutane pyrimidine dimers. AB - DNA repair defects can predispose to cancer development and progression. We previously showed that the breast and ovarian cancer susceptibility gene product BRCA1, through p53, upregulates expression of the XPE gene DDB2 encoding the nucleotide excision repair protein p 48. Both XPE and XPC are p53 target genes containing p53 response elements. To further explore the role of BRCA1 and p53 in repair of photoproducts, we eliminated wild type p53 from U2OS osteosarcoma cells and found that cyclobutane pyrimidine dimer (CPD) repair was markedly impaired following UV damage whereas repair of 6-4 photoproduct (6-4 PP) occurred efficiently. Overexpression of p53 in p53-null Calu-6 cells also enhanced CPD repair. In HCC1937 breast cancer cells, harboring mutant BRCA1 and p53 genes, repair of CPD was markedly impaired. Reintroduction of either p53 or BRCA1 using adenovirus vectors into HCC1937 alone had little effect on repair of CPD whereas the combination of p53 and BRCA1 resulted in efficient repair of CPD. Thus there appears to be a cooperative effect between p53 and BRCA1 that may involve induction of repair proteins, inhibition of p53-induced cell death by BRCA1 with altered p53 selectivity towards repair pathways and/or p53-independent effects of BRCA1 on CPD repair. PMID- 16357512 TI - Phase I clinical trial of oral 2-methoxyestradiol, an antiangiogenic and apoptotic agent, in patients with solid tumors. AB - PURPOSE: To determine the maximum-tolerated dose (MTD) and toxicity profile of the novel anticancer agent, 2-methoxyestradiol (2ME2) administered orally, in patients with solid tumors. MATERIALS AND METHODS: Twenty patients with refractory solid tumors were enrolled. 2ME2 was given orally starting at 400 mg bid with dose escalation until 3000 mg bid. Tumor biopsies were taken before and after starting the drug to assess for microvessel density by CD 31 and cell proliferation by Ki67 immunohistochemistry. Serial plasma samples collected up to 50 hours after first single oral dose for characterization of pharmacokinetics, were analyzed using liquid chromatography tandem mass-spectrometry. RESULTS: Eleven men and nine women received 2ME2 at dose levels of 400 mg bid (n = 3), 800 mg bid (n = 3), 1600 mg bid (n = 6), 2200 mg bid (n = 5) and 3000 mg bid (n = 3). There were no dose limiting toxicities, therefore the MTD was not defined. There was one episode of grade 4 angioedema in the 1600 mg bid dose level 38 days into 2ME2 treatment. Other toxicities were mild to moderate. A patient with clear cell carcinoma of the ovary had a partial response at 1600 mg bid dose level lasting over three years. CONCLUSION: MTD for 2ME2 was not reached at dose of 3000 mg bid. The trial was closed due to extremely low plasma concentrations of 2ME2 relative to the doses administered. 2ME2 treatment had no effect on microvessel density (CD31 immunostaining) and cell proliferation (Ki-67 immunostaining). A new formulation of 2ME2 with improved bioavailability is currently being developed. PMID- 16357513 TI - Paclitaxel encapsulated in cationic lipid complexes (MBT-0206) impairs functional tumor vascular properties as detected by dynamic contrast enhanced magnetic resonance imaging. AB - Cationic lipid complexes have been shown to be bound and internalized selectively by angiogenic tumor endothelial cells after intravenous injection. Based on this phenomenon, the chemotherapeutic agent paclitaxel was encapsulated into these lipid complexes providing a vascular targeting agent (MBT-0206). As noninvasive imaging techniques are of critical importance for optimizing antivascular cancer treatment in the clinic, we have evaluated the antivascular effects of MBT-0206 in the A-MEL-3 solid tumor model using dynamic contrast enhanced magnetic resonance imaging (DCE-MRI). Twenty-four hours after three intravenous applications of MBT-0206, tumors of treated animals demonstrated a significant decrease of intratumoral blood volume and an increase of vascular permeability in comparison to size-matched control tumors. In contrast, animals treated with conventional paclitaxel given as Taxol at equal drug dose did not show any significant differences in vascular parameters acquired by DCE-MRI in comparison to controls. Immunohistological analysis confirmed a significant reduction of microvessel density in MBT-0206 treated tumors. Moreover, a significant increase of intratumoral microvascular occlusion following MBT-0206 treatment was observed compared to controls and paclitaxel treated animals. In conclusion, antivascular tumor therapy with MBT-0206 significantly impairs functional tumor microcirculation. DCE-MRI is a promising tool to quantify the antivascular effects of MBT-0206 during treatment. PMID- 16357514 TI - Cytotoxicity of a natural anthraquinone (Aloin) against human breast cancer cell lines with and without ErbB-2: topoisomerase IIalpha coamplification. AB - In the present study the cytotoxic activity of aloin, a natural anthracycline from Aloe plant, is reported against two human breast cancer cell lines; without (MCF-7) and with (SKBR-3) erbB-2-topoIIalpha coamplification. MCF-7cell line was shown to be more sensitive to aloin than SKBR-3 demonstrated by MTT and clonogenic assays, from which IC50 and 50% ICF values are reported to be 60 microg/ml, respectively, in the former cell line and as high as 150 and 80 microg/ml, respectively, in the latter, which are still far below the maximum tolerated dose of the compound. The effect of aloin is suggested to be brought about by more than one mechanism depending on the dose level and tumor phenotype. This was demonstrated by flow cytometric analysis, fluorescence microscopy and western blot analysis, which revealed that aloin at higher concentrations caused a reduction in the proportion of cells undergoing mitosis by induction of apoptosis, inhibition of topo II alpha protein expression and downregulation of cyclin B1 protein expression in MCF-7 cell line, whereas erbB-2 protein expression was not affected. Topo IIalpha protein expression was mildly downregulated in SKBR-3 cell line at higher concentrations only. PMID- 16357515 TI - CDC4 gene expression as potential biomarker for targeted therapy in prostate cancer. AB - Targeted therapy of proteasome regulated gene expression has potential utility in cancer treatment since components of ubiquitin-mediated proteolysis are altered in human malignancy. Specific regulators of proteasome degradation such as F-box proteins of the SCF E3 ligase complex are ideal biomarkers for assessing therapeutic efficacy since these components determine substrate specificity. An F box protein that appears to be important in this process is human Cdc4 (Fbw7) since expression is detected in a variety of human cancers including breast, colon, pancreas and uterus. The role of Cdc4 in tumorigenesis appears to be related at least in part to regulation of cyclin E since inactivating mutations of CDC4 in cancer cells leads to cyclin E overexpression and genomic instability. In order to investigate the potential biological and clinical consequences of proteasome inhibition with respect to Cdc4 mediated targeted proteolysis, we investigated CDC4 expression and genetic alterations in 53 primary human prostate cancers in addition to correlation with relevant histopathological and clinical parameters. We identified genetic alterations in 6% of our prostate cancers while differential expression of Cdc4 isoforms correlated with advanced pathological stage and clinical recurrence. Our data suggest that CDC4 expression in prostate cancer has important biological and clinical implications since genetic alterations, differential Cdc4 isoform expression, histopathological and clinical correlation were demonstrated in our analysis. Therefore molecular genetic analysis of CDC4 expression may be an important biomarker for concurrent or subsequent clinical investigation of proteasome targeted therapy in men with prostate cancer. PMID- 16357516 TI - Such an obsession. PMID- 16357517 TI - pRb2/p130 and VEGF expression in endometrial carcinoma in relation to angiogenesis and histopathologic tumor grade. AB - PURPOSE: Endometrial cancer is the most common gynecologic malignancy. Established prognostic factors are histologic grade, depth of myometrial invasion, and extrauterine spread including retroperitoneal lymph node metastases. Tumorigenesis is a multistep process involving different genetic changes resulting in uncontrolled cellular proliferation, inhibition of apoptosis, and enhanced vascular proliferation among other events. Angiogenesis, the formation of new blood vessels from a preexisting vascular network, is necessary for invasive tumor growth and metastasis and constitutes an important point in the control of cancer progression. The pathogenesis of the angiogenetic phenotype may involve the inactivation of different tumor suppressor genes. EXPERIMENTAL DESIGN: We investigated the relationship between the expression levels of VEGF and the retinoblastoma family member pRb2/p130 in endometrial carcinoma in relation to histopathologic tumor grade in a cohort of 50 patients. RESULTS: We found that VEGF and pRB2/p130 expression were inversely correlated. Additionally, high grade tumors presented a significantly lower number of cells expressing pRb2/p130 when compared to low grade tumors. A significant positive correlation was found, by means of the Spearman coefficient, between VEGF expression and binary grading (0.450, p-value < 0.005) which is an architectural grading system that uses low-magnification assessment of amount of solid growth, pattern of invasion, and presence of necrosis to divide endometrioid carcinomas into low- and high-grade tumors. Additionally, we also found a negative correlation between pRb2/p130 expression levels and binary grading (-0.595, p value < 0.005). Interestingly, we also found that VEGF and pRb2/p130 expression levels were not related to staging (p-value > 0.005). CONCLUSIONS: These results open up a new perspective including novel markers that, combined together, may be useful in patient screening for endometrial cancer aggressiveness. PMID- 16357518 TI - Differential gene expression profiling in aggressive bladder transitional cell carcinoma compared to the adjacent microscopically normal urothelium by microdissection-SMART cDNA PCR-SSH. AB - Identifying novel and known genes that are differentially expressed in aggressive bladder transitional cell carcinoma (BTCC) has important implications in understanding the biology of bladder tumorigenesis and developing new diagnostic and therapeutic agents. In this study we identified the differential gene expression profiles comparing tumor to the adjacent microscopically normal mucosa by manual microdissection on frozen sections. The RNAs extracted from microdissected tissues were amplified by SMART cDNA PCR technology to generate forward subtractive cDNA library by suppressive subtractive hybridization (SSH). We obtained 376 positive clones, one hundred clones of aggressive BTCC subtracted cDNA library were selected at random and inserts were reamplified by PCR. After differential screening by reverse dot blotting, 73 positive clones, that contend inserts putatively upregulated in aggressive BTCC, were further analysed by DNA sequencing, GenBank and EST database searching. Sequencing results showed that 66 clones stand for 23 known genes and 7 clones for three new EST (Genbank number: DN236875, DN236874 and DN236873). In conclusion, microdissection-SMART cDNA PCR SSH allowed for an efficient way to identify aggressive BTCC-specific differential expressed genes that may potentially be involved in the carcinogenesis and/or progression of aggressive BTCC. These differentially expressed genes may be of potential utility as therapeutic and diagnostic targets for aggressive BTCC. PMID- 16357519 TI - Insulin-like growth factor binding protein-2 stimulates proliferation and activates multiple cascades of the mitogen-activated protein kinase pathways in NIH-OVCAR3 human epithelial ovarian cancer cells. AB - Insulin-like growth factor binding protein-2 (IGFBP-2), the second most abundant IGFBP in the circulation, is dramatically increased in the serum and ovarian cyst fluid of women with epithelial ovarian cancer. The specific role of IGFBP-2 in ovarian carcinogenesis remains elusive. Using NIH-OVCAR3 human epithelial ovarian cancer cells, we have evaluated the effects of IGFBP-2 and its antibody on cell proliferation, activation of mitogen-activated protein kinase (MAP kinase) pathways and on cytokine expression. Treatment of the cells with IGFBP-2 stimulates cell growth significantly (p<.05) and potentiates the activation of (1) the extracellular signal regulated kinase (ERK1/2) signaling pathway, which transduces cell-specific growth and differentiation signals; (2) the stress activated protein kinases/c-Jun N-terminal kinases (SAPK/JNK) pathway, which is activated by environmental stresses, inflammatory cytokines, growth factors and G protein coupled receptor (GPCR) agonists; and (3) the p38 MAP kinase pathway, which mediates inflammatory and stress responses. Suppression of IGFBP-2, with its neutralizing antibody, significantly (p<.05) retards cell growth, blocks the activation of all three cascades of the MAPK pathways and downregulates the expression of a number of potential cancer-promoting cytokines. These novel findings may have important clinical implications for developing innovative strategies for the treatment and management of ovarian cancer. PMID- 16357520 TI - Gefitinib-sensitizing mutation in esophageal carcinoma cell line Kyse450. AB - PURPOSE: The sensitivity of lung cancer to gefitinib has been found to be associated with mutations at the tyrosine kinase domain of epidermal growth factor receptor (EGFR), yet similar observations are not available in other solid tumors. We recently identified mutations in the EGFR kinase domain in primary esophageal carcinoma. The purpose of this study was to investigate if they are gefitinib-sensitizing EGFR mutations. EXPERIMENTAL DESIGN: We identified a missense mutation in the EGFR kinase domain, EGRFS7681, in the esophageal cancer cell line Kyse450. The sensitivity of this cell line to gefitinib was compared to an esophageal cancer cell line with wildtype EGFR, TE8, and to a lung cancer cell line, H358, known to be resistant to gefitinib. The effect of EGFR(S7681) mutation on cell growth and apoptosis was assessed. RESULTS: As demonstrated by in vitro proliferation assay, this mutation sensitized Kyse450 cells to gefitinib. The observation of down regulation of the phosphorylated pAKT indicated gefitinib induced Kyse450 cells apoptosis via inhibition of EGFR activity CONCLUSIONS: While more primary esophageal tumors remain to be screened for the mutations at the tyrosine kinase domain of EGFR, current observation implies that gefitinib may be worth further investigation for treatment of esophageal cancers. PMID- 16357521 TI - ENU administration causes genomic instability along with single nucleotide polymorphisms in p53 during gliomagenesis: T11TS administration demonstrated in vivo apoptosis of these genetically altered tumor cells. AB - Advancement of molecular analysis of neoplastic cells demonstrated that multiple genetic changes are associated with the development of tumors. Cancer cell must exhibit a mutator phenotype, which is likely to be responsible for the genomic instability found in cancer tissues. The mutator phenotype, such as defective mismatch repair, is known to cause microsatellite instability, which is associated with certain cases of sporadic cancer. Previously many studies have been carried out to determine the relationship between microsatellite instability and human brain tumors. However information on genomic instability in the animal model of brain tumor is still very limited. In the present course of investigations we genetically characterized our ENU induced brain tumor animal model by using PCR based randomly amplified polymorphic DNA (RAPD) analysis and restriction fragment length polymorphism (RFLP) with three microsatellite probes. ENU induced tumors demonstrated genetic instability, including some microsatellite instability. As single nucleotide polymorphisms of the tumor suppressor gene p53 were associated with diverse types of human cancer, we examined the p53 gene of the tumor cells isolated from ENU induced brain tumor animal model, by PCR based RFLP method in p53 exon-2, -3 and -4. In these studies we showed that the restriction site of p53 exon-3 and 4 were mutated in ENU induced brain tumor indicating a genetic defect associated with ENU induced tumorigenesis. In the therapeutic part, we confirmed the anti-tumor property of T11TS/S-LFA-3 in the ENU induced genetically altered cells. Histological evidences, cytotoxic study, PI-FACS cell-cycle analysis and TUNEL assay confirmed the apoptotic death of glioma cells by T11TS treatment in which p53 is mutated. From the present study we can conclude that ENU administration causes genomic instability along with mutations in p53 during the process of gliomagenesis. Whereas, T11TS/S-LFA3 demonstrated the potential to induce apoptosis of these tumor cells even when p53 is mutated and thus showed its immense potential to be an anti-neoplastic probe against p53 mutated diverse types of tumors. PMID- 16357522 TI - Treatment of NSCLC patients with an EGF-based cancer vaccine: report of a Phase I trial. AB - Epidermal Growth Factor (EGF) promotes tumor cell proliferation and survival upon binding to its receptor. We have developed a new active specific immunotherapy based on EGF deprivation. In the present paper, we show the results of a Phase I trial in 43 patients with advanced non-small cell lung cancer (NSCLC) who received the EGF vaccine. Patients who had already received first line therapy were randomized to receive a single or double dose of the EGF vaccine, weekly for four weeks and monthly thereafter. No significant toxicity was seen after vaccination. Adverse events consisted primarily of fever, chills, nausea, vomiting and flushing. Fifteen patients (39%) developed a good antibody response (GAR) against EGF. The geometric mean of the antibody titer was higher in the double dose group. EGF concentration was quantified in serum. An inverse correlation between anti-EGF antibody titers and EGF concentration was seen after immunization. Vaccinated patients achieved median survival times of 8.23 months from randomization. Patients who received the double dose of treatment showed a trend toward increased survival in comparison with patients who received the single dose. GAR and patients in whom the serum EGF decreased below the 168 pg/ml cut-off point had a significantly better survival when compared to poor responders or patients in which the EGF levels were not considerably reduced. Our results confirm the immunogenicity of the EGF vaccine in the treatment of patients with advanced stage NSCLC. Antibody titers and serum EGF levels appear to correlate with patient survival. PMID- 16357523 TI - Production and post-surgical modification of VEGF, tPA and PAI-1 in patients with glioma. AB - Malignant gliomas are associated with risk of thromboembolism, but the molecular link between tumor and peripheral pro-coagulant status has not been elucidated. Vascular Endothelial Growth Factor (VEGF), tissue-type Plasminogen Activator (tPA), Plasminogen Activator Inhibitor-1 (PAI-1) and lipoprotein (lp) (a) influence the pro-coagulant status. To assess whether the presence of the tumor influenced the peripheral levels of VEGF, tPA, PAI-1 and lp(a), we studied the expression and secretion of VEGF, tPA, PAI-1 and lp(a) in glioma specimens, in peripheral blood and in primary glioma-derived cultures. We also measured lp(a), VEGF, tPA and PAI-1 in the peripheral circulation of patients, before and after surgery for glioma. VEGF, tPA and PAI-1 were expressed in glioma specimens. Glioma cells were indeed a major source of tPA and PAI-1; these molecules were significantly more expressed in glioma than in patient's blood cells. Lp(a) was rarely expressed in glioma specimens and not expressed in blood cells. In glioma, VEGF, tPA and PAI-1 were localized mainly in tumor cells; tPA was localized also in the extracellular matrix and PAI-1 in tumor vascular lumen. Glioma cells were indeed able to produce and release VEGF, tPA and PAI-1. After surgery, peripheral levels of VEGF and PAI-1 were increased, while tPA and lp(a) were unchanged. The great amount of VEGF, tPA and PAI-1 produced by glioma could influence peripheral levels of these molecules. The partial resection of the tumor by surgery was not able to decrease plasma levels of these molecules. PMID- 16357524 TI - Adenovirus E1A reverses the resistance of normal primary human lung fibroblast cells to TRAIL through DR5 upregulation and caspase 8-dependent pathway. AB - Expression of the adenovirus serotype 5 (Ad5) E1A enhances tumor cells to apoptosis by TNF-alpha, Fas-ligand and TNF-related apoptosis-inducing ligand (TRAIL). In this study, we found that E1A expression reversed the resistance of normal primary human lung fibroblast cells (P-HLF) to TRAIL-induced apoptosis. Furthermore, TRAIL dramatically induced apoptosis of P-HLF cells that expressed E1A following either infection with Ad-E1A or transfection with pcDNA3-E1A. Further results demonstrated that E1A specifically upregulated DR5 levels but had nearly no effect on the levels of DR4. E1A dramatically upregulated the exogenous TRAIL, and then increased a substantial amount of TRAIL on the surface of P-HLF cells treated with the expression vectors, both Ad-TRAIL and pIRES-EGFP-TRAIL. The dominant negative FADD mutation (FADD-DN) results revealed that the apoptosis in Ad-E1A and Ad-TRAIL coinfected P-HLF cells was completely blocked following inhibition of the death receptors-associated apoptosis-inducing molecules FADD. Moreover, the caspase 8 inhibitor (Z-IETD-FMK) could efficiently block caspase 8 activation and resulted in inhibition of caspase 3 activation and cleavage. However, The caspase 9 specific inhibitor (Z-LEHD-FMK) could not counteract the synergistic effect of TRAIL-induced apoptosis in combination with E1A, and caspase 3 activation and cleavage were not inhibited by Z-LEHD-FMK. Thus, our results suggest that adenovirus E1A sensitizes P-HLF cells to TRAIL-induced apoptosis involving DR5 upregulation and the caspase 8-dependent pathway. These findings provide the first direct evidence for molecular mechanisms of adenovirus E1A gene products to sensitize normal cells to TRAIL-mediated apoptosis. PMID- 16357525 TI - The diverging roles of calmodulin and PKC in the regulation of p21 intracellular localization. AB - Intracellular localization plays an important role in the functional regulation of the cyclin-dependent kinase inhibitor p21. While nuclear functions have been linked to the tumor suppressor activity of p21, cytoplasmatic functions are oncogenic. We have recently shown that Ser153 phosphorylation of p21 by PKC contributes to its cytoplasmatic accumulation, and that this phosphorylation is inhibited by Ca(2+)-dependent calmodulin binding to the C-terminal region of p21. Consequently, PKC and calmodulin/Ca(2+) play diverging roles in the regulation of p21 intracellular localization. Other kinases such as AKT and MIRK/dyrk1B also phosphorylate p21 near the nuclear localization signal, thus inhibiting its nuclear accumulation. We discuss here the effects of such phosphorylations on p21 functionality, as well as its relevance to cell cycle progression and differentiation. PMID- 16357526 TI - Mitochondrial impairment is a critical event in anthrax lethal toxin-induced cytolysis of murine macrophages. AB - Numerous early events in anthrax lethal toxin (LT)-mediated cell killing have been described, including uptake of LT and MAPKK cleavage. However, critical downstream events in LT killing remain to be identified. In this study we present evidence that LT causes mitochondrial dysfunction in murine J774A.1 macrophages, as indicated by a continuous drop in both mitochondrial membrane potential and SDH activity. This was further supported by ultrastructural analysis revealing LT induced swelling of mitochondria. Mitochondrial impairment and cytolysis were controlled by proteasomes in LT-treated macrophages: proteasome inhibitors restored mitochondrial activity and rescued cells from cytolysis, even when added immediately prior to membrane perturbation. Similar to proteasome inhibitors, KCl also efficiently blocked LT-mediated cytolysis, even after late addition. However, KCl did not prevent mitochondrial impairment, though it precluded events linked to LT-induced cytolysis. These events included a precipitous drop in ATP levels and ubiquitinated proteins, revealing that they are epiphenomena in LT killing. Our studies suggest that proteasomes and potassium control LT-induced mitochondrial dysfunction and membrane perturbation, key events in LT killing. PMID- 16357527 TI - R-Spondin proteins: a novel link to beta-catenin activation. AB - The R-spondin (Rspo) protein family is a recently described group of four distinct human secreted proteins. Reported activities for Rspo proteins include essential roles in vertebrate development and their ligand-type activities overlap substantially with those of the canonical Wnt ligands in that both Rspo and canonical Wnt signaling result in the activation of beta-catenin. In a general functional screen for human secreted proteins using transgenic mouse models, we identified human R-spondin1 (hRspo1) protein as a potent and specific mitogen for the gastrointestinal epithelium and demonstrated a potential therapeutic application for the protein in mouse models of cancer therapy-induced mucositis. In contrast to previous studies, our data indicated only partial overlap between Wnt and Rspo ligand activities, suggesting that there may be independent receptor/signaling pathways for Rspo proteins that intersect those of Wnt at the level of beta-catenin. Here we summarize the current reported data on the Rspo family and discuss these results in terms of alternate mechanisms of action. We have extended our observations on the potential therapeutic application of Rspo proteins by showing that all four human Rspo family members are capable of inducing epithelial proliferation and report the first non vertebrate Rspo family member. PMID- 16357528 TI - HOXB4 inhibits cell growth in a dose-dependent manner and sensitizes cells towards extrinsic cues. AB - Ectopic expression of the homeodomain transcription factor HOXB4 expands hematopoietic stem and progenitor cells in vivo and in vitro, making HOXB4 a highly interesting candidate for therapeutic stem cell expansion. However, when expressed at high levels, HOXB4 concomitantly perturbs differentiation and thus likely predisposes the manipulated cells for leukemogenesis. We therefore asked whether the expression level of HOXB4 may be a critical parameter that influences the growth and transformation properties of transduced cells. Using a set of retroviral vectors which covered a 40-fold range of expression levels, we studied the consequences of HOXB4 expression at different levels in the well established Rat-1 fibroblast cell system. HOXB4 transformed Rat-1 fibroblasts beyond a certain threshold level of expression. Further escalation of HOXB4 expression, however, did not enhance transformation. Instead, HOXB4 mediated a dose dependent anti-proliferative effect on Rat-1 and NIH3T3 fibroblasts. This effect was aggravated under reduced serum concentrations and was, at least partially, due to an enhanced sensitivity of HOXB4 overexpressing cells to induction of apoptosis. Based on these results we propose that HOXB4 affects cell growth in a dose dependent manner by sensitizing cells towards extrinsic signals. PMID- 16357529 TI - Assessing the link between BACH1 and BRCA1 in the FA pathway. AB - The BACH1 helicase was initially identified by its direct binding to BRCA1 and, thus, was linked to hereditary breast cancer. More recently, BACH1 was identified as the gene defective in the J complementation group of Fanconi anemia (FA). FA is a multigenetic disorder characterized by cellular sensitivity to crosslinkers and chromosome instability. Because FANCD2 monoubiquitination is intact in BACH1 deficient cells, BACH1 appears to act downstream in the FA pathway akin to BRCA2/FANCD1. Interestingly, while BRCA1 has various interactions with FA proteins it has not been identified as an FA gene. As the race to uncover the last few unknown FA complementation groups comes to an end, future work will be required to uncover how these gene products function to combat the effects of DNA damage and maintain genomic stability. In particular, it remains elusive whether BRCA1 is functionally linked to the FA pathway through its interaction with BACH1/FANCJ. This review focuses on a model for the connection of BRCA1 to BACH1 in the FA pathway. We predict that BRCA1 regulates the BACH1 helicase activity to coordinate the timely displacement of Rad51 from nucleofilaments, promoting error free repair and ultimately maintaining chromosomal integrity. PMID- 16357530 TI - Less REST, more vascular disease? Regulation of cell cycle and migration of vascular smooth muscle cells. AB - Neointimal hyperplasia and atherosclerosis are cardiovascular abnormalities resulting in heart attacks and strokes--major causes of death and disability in societies with a western-style diet. Both are contributed to significantly by a phase of enhanced cell cycle activity and migration of vascular smooth muscle cells. Here we describe how this activity is associated with a switch in the types of ion channel controlling trans-membrane ion transport. In particular we stress the de novo expression of the K(Ca)3.1 potassium channel when the cells proliferate, but we also review evidence of changes in other ion channels. The molecular mechanisms governing such switches in ion transport have been unknown until we recently showed downregulation of the repressive REST transcription factor is an important step. Here we explain and expand the hypothesis that REST regulation of ion transport has a previously unrecognised role in cell proliferation. PMID- 16357531 TI - Control of hepatic differentiation by activin/TGFbeta signaling. AB - During liver development, liver progenitors called hepatoblasts differentiate into hepatocytes or biliary cells. Recently, we showed that the segregation between hepatocytes and biliary cells is dependent on a gradient of Activin/TGFbeta signaling, and that Activin/TGFbeta signaling is controlled in fetal liver by transcription factors of the Onecut family. Here, we discuss candidate factors possibly involved in the formation of the Activin/TGFbeta signaling gradient, how this gradient could integrate into a network of signaling pathways modulating hepatoblast differentiation, and the implications for human liver disease and therapy. PMID- 16357532 TI - Regulation of Separase in meiosis: Separase is activated at the metaphase I-II transition in Xenopus oocytes during meiosis. AB - Separase is a cysteine protease conserved in all eukaryotes and functions to remove sister chromatid cohesion in anaphase by cleaving the SCC1 subunit of the cohesin complex. Separase activity is regulated by its inhibitor securin and by an inhibitory phosphorylation in vertebrates. However, these regulations have never been directly investigated in the meiotic cell cycle of vertebrates. In this study, we cloned the full-length gene encoding Xenopus separase from an oocyte cDNA library. Purified xSeparase can cleave the human alpha-kleisin subunit of cohesin in vitro but cannot bind to hSecurin when these two proteins are coexpressed in 293T cells. Similar to its human counterpart, xSeparase cleaves itself upon activation but at a single site. The cleavage site is conserved with one of the three self-cleavage sites in hSeparase. Using self cleavage as a reporter for its activation, we demonstrated that xSeparase is transiently activated between the two meioses and may be involved in homolog separation, as is observed in other organisms. Taking advantage of the inability of xSecurin to interact with hSeparase, we demonstrated that CSF extract can reinhibit both full-length and auto-cleaved hSeparase, indicating that inhibition of separase by phosphorylation does occur under physiological conditions. In addition, we found that endogenous xSecurin accumulated in response to progesterone-induced oocyte maturation and was degraded at both anaphase I and II in an APC/C-dependent manner. PMID- 16357533 TI - Brain tumors provide new clues to the source of cancer stem cells: does oncology recapitulate ontogeny? AB - Recent studies of leukemia and solid tumors have provided compelling evidence for the existence of cancer stem cells, but the origin of these cells remains a matter of considerable debate. By comparing the gene expression profiles of ependymomas with those of cells in the normal developing nervous system, we were able to pinpoint radial glia as candidate stem cells of this brain tumor. These data suggest strongly that ependymomas arise directly from transformed radial glia and they provide a novel method that could be used to map the cell of origin of other types of cancer. PMID- 16357534 TI - Dual-color-coded imaging of viable circulating prostate carcinoma cells reveals genetic exchange between tumor cells in vivo, contributing to highly metastatic phenotypes. AB - Color-coded imaging analysis revealed yellow fluorescent metastasis precursor cells that were readily recognized in the blood of tumor-bearing mice after mixtures of red fluorescent protein (RFP)- and green fluorescent protein (GFP) expressing PC-3 human prostate carcinoma cells were implanted in the nude mouse prostate and metastasized. The yellow fluorescent cells were purified from the blood of nude mice to 99% homogeneity by FACS, expanded in culture, and reimplanted in the prostate of nude mice. The yellow fluorescent phenotype was heritable and stably maintained by tumor cells for many generations in vitro and in vivo. In the animals implanted with the yellow-fluorescing cells, 100% developed aggressive metastatic cancer. Lung metastases were demonstrated in 100% of the animals as early as four weeks after injection of the yellow-fluorescing cells in the mouse prostate. In contrast, when the GFP- and RFP-expressing parental cells were inoculated into the mouse prostate separately, none of the animals developed lung metastasis. All animals had almost exclusively yellow fluorescent cells in the blood and bone marrow. These results are consistent with the idea that spontaneous genetic exchange between tumor cells in vivo contributes to genomic instability and creation of highly metastatic cells. PMID- 16357535 TI - Mammalian gene expression program resiliency: the roles of multiple coactivator mechanisms in hypoxia-responsive transcription. AB - CBP and its paralog p300 (CBP/p300 collectively) are transcriptional coactivators that are among the most interconnected proteins in the mammalian protein-protein "interactome" with over 315 described interaction partners. CBP/p300 are protein/histone acetyltransferases, but most of the protein-binding domains of CBP/p300 are unique to these two coactivators, indicating that CBP/p300 should be highly limiting. The CH1 domain of CBP/p300 was considered essential for most, if not all, hypoxia-inducible transcription by binding to hypoxia-inducible-factor 1alpha (HIF-1alpha). Mutating CH1 had little effect, however, on the hypoxia induced transcription of the HIF-target genes Higd1a, Egln1 (prolyl-hydroxylase), Bnip3 (Bcl2-interacting-protein-3), and Pfkl (phosphofructokinase). In contrast, HIF-targets Pgf (placental growth factor) and Egln3 were strongly affected by the CH1 mutation, while Stc1 (stanniocalcin-1) and Slc2a1 (glucose-transporter-1) were moderately affected. HIF targets were also dependent on coactivation mechanisms that are sensitive to trichostatin A (TSA(S)). Paradoxically, TSA inhibits histone deacetylases (HDACs) that are usually associated with transcriptional repression, implying that HDACs can also function as coactivators. Thus, activator-specific transcription in mammals requires seemingly unrelated coactivator mechanisms, and individual target genes vary in their requirements for each mechanism. Gene expression program resiliency is therefore coupled with gene specific regulation by avoiding uniform reliance on a "keystone" coactivator interaction. PMID- 16357536 TI - CUTL1: a key mediator of TGFbeta-induced tumor invasion. AB - The TGFbeta pathway plays a dual role in human carcinogenesis. On one hand, TGFbeta is well known for its ability to inhibit epithelial cell proliferation and promote apoptosis. However, many advanced cancers acquire resistance to the growth-inhibitory effects of TGFbeta and respond to it instead with promotion of proliferation, invasion and tumor progression. The homeobox transcription factor CUTL1, also known as CCAAT displacement protein, CDP or Cux-1, is involved in the control of normal embryonic development and differentiation. Recently, we found that CUTL1 is a transcriptional target of TGFbeta and is an important mediator of the TGFbeta-induced cell migration and invasion. In addition, CUTL1 is highly expressed in various epithelial cancers and seems to negatively correlate with tumor differentiation and patient survival. Therefore we postulate that CUTL1 might be a key mediator of the tumor-promoting effects of TGFbeta in advanced cancers. PMID- 16357537 TI - Exploiting the compromised spindle assembly checkpoint function of tumor cells: dawn on the horizon? AB - Aneuploidy is a frequent property of cancer cells that arises as a consequence of chromosomal instability (CIN). A major safeguard mechanism protecting cells from CIN is the spindle assembly checkpoint. This checkpoint surveys proper attachment of chromosomes to the mitotic spindle and effectively suppresses erroneous chromosome segregation by delaying mitotic progression until proper spindle chromosome interactions have been established. Several lines of evidence suggest that the development of aneuploidy may be a gradual process that in many cases could result from subtle spindle checkpoint defects that occur during tumorgenesis and steadily weaken spindle checkpoint function. Here we discuss the evidence for this concept and address the question whether normal somatic cells and tumor cells could perhaps exhibit differences in spindle checkpoint regulation that allow the design of more specific anti-tumor strategies that effectively kill tumor cells but spare normal cells. PMID- 16357538 TI - The MicroRNA pathway plays a regulatory role in stem cell division. AB - One of the key characteristics of stem cells is their capacity for self-renewal for long periods of time. In this respect, stem cells are similar to cancer cells, which also have the ability to escape cell cycle stop signals. Therefore, a critical question in stem cell and cancer biology is how cell division is regulated in these cell types. In this review, we summarize recent progress and describe future challenges to understanding the role the microRNA pathway plays in regulating mechanisms controlling stem cell division. PMID- 16357539 TI - Defining cell lineages in the prostate epithelium. AB - Understanding the stages of cell differentiation in the normal prostate epithelium is essential for the identification of the cell type(s) involved in prostatic carcinogenesis. Prostate glands are composed of three types of epithelial cells (i.e., basal, secretory and neuroendocrine) but the hierarchical relations among these cell types have been long controversial. We have recently developed a novel system to define prostate epithelial cell lineages in vivo. We find that, during normal prostate organogenesis, terminally differentiated secretory cells derive from p63-positive basal cells, which thus represent/include prostate stem cells. Future studies will determine if p63 positive basal cells retain stem cells capabilities in the adult prostate epithelium. PMID- 16357541 TI - Turning the replication checkpoint on and off. AB - The replication checkpoint monitors the progress of DNA replication forks during S phase, and delays the firing of later replication origins when active replication forks are stalled due to collisions with damaged or abnormally structured DNA. Key components of the replication checkpoint pathway are the apical protein kinase, ATR, and its downstream target kinase, Chk1. Defects in either ATR or Chk1 function result in loss of DNA replication fidelity and cell viability, even in the absence of extrinsic genotoxic stress. Moreover, several clinically important antitumor agents, such as the camptothecins (CPTs), exert their antitumor effects by interfering with DNA replication, and hence the therapeutic response to these drugs is intimately related to signaling through the replication checkpoint. A recent report from this laboratory adds a new facet to the regulatory mechanisms that control the function and duration of checkpoint signaling through the ATR-Chk1 pathway. The results indicate that replication stress induced by a variety of agents, including CPT and deep hypoxia, triggers the ubiquitin-dependent degradation of the checkpoint kinase Chk1 in both normal and transformed human cells. This review provides an overview of the study's major findings, together with their implications for both replication checkpoint function and tumor responsiveness to CPT and related anticancer drugs. PMID- 16357540 TI - Differential regulation of D-type cyclins in the mouse intestine. AB - The intestinal epithelium undergoes continuous rapid renewal throughout adult life. To examine contributions of D-type cyclins to proliferation in the intestine, we examined D-type cyclin expression in the mouse proximal and distal small intestine and colon. Cyclin D1 was expressed throughout the small and large intestine. In contrast, cyclin D2 and D3 protein levels were not readily detectable in the duodenum. Levels of RNAs encoding all three D-type cyclins were higher in the ileum and colon than in the duodenum. Immunohistochemistry revealed that cyclin D1 and D2 are expressed in colonic epithelial cells, with cyclin D2 being more restricted to the proliferative zone. Expression of cyclinD1 and D2 was detected in conditionally immortalized young adult mouse colon (YAMC) cells and in a colon epithelial cell line derived from the Apc(Min/+) mouse (IMCE cells), with higher basal levels of both cyclins in the IMCE cells. In an experimental model of colitis, levels of cyclin D1 mRNA increased significantly, and cyclin D1 protein was localized to both epithelial cells and inflammatory cells in the colon. The individual D-type cyclins may make different contributions to proliferation, disease and development of cancer in the intestine. PMID- 16357542 TI - Outcome after rehabilitation for total joint replacement at IRF and SNF: a case controlled comparison. AB - OBJECTIVE: The objective of this study was to determine whether outcomes differed between patients with single knee or hip joint replacement surgery undergoing rehabilitation in an inpatient rehabilitation facility (IRF) vs. skilled nursing facilities (SNFs). DESIGN: A retrospective chart review was performed on 87 pairs of patients treated in either an IRF or a SNF matched for age, gender, type of surgery, and Functional Independent Measure (FIM) motor score at admission. All patients discharged from the IRF for rehabilitation following single hip or knee replacement surgery in 2004 were eligible for comparison with index cases discharged from SNFs with the same diagnosis in 2004. At discharge, FIM motor scores, device used for ambulation, ambulation distance, disposition, and length of stay (LOS) were recorded. RESULTS: The mean LOS of IRF-treated patients was 10.3 +/- 3.3 days, compared with 20.0 +/- 10.8 days for SNF-treated patients (P < 0.005). A significantly higher percentage of IRF-treated patients were discharged directly home (IRF: 89.5%; SNF: 79.1%; P < 0.029). The mean discharge locomotion FIM score for IRF-treated patients was 5.71 +/- 0.91 compared with 4.90 +/- 1.92 for the SNF-treated patients (P < 0.004). At discharge, the mean ambulation distance of patients treated at the IRF was of 380 +/- 168 feet compared with 289 +/- 212 feet for patients treated at SNFs (P < 0.005). Significantly more of the SNF-treated patients required a walker (80.2%) for ambulation at discharge compared with patients treated at the IRF (38.3%, P < 0.001). Of the patients who were discharged home, 75% of the SNF-treated patients required homecare services compared with 41.2% of the IRF-treated patients (P < 0.001). CONCLUSION: When patients were matched for age, gender, operative diagnosis, and admission ambulation FIM, those who received rehabilitation in the IRF had, on average, a shorter length of stay and superior functional outcomes than those treated in SNFs. PMID- 16357543 TI - Side effects and complications after percutaneous disc decompression using coblation technology. AB - OBJECTIVE: To report the short-term side effects and complications after percutaneous disc decompression utilizing coblation technology. DESIGN: Following institutional review board approval, consecutive patients who were to undergo percutaneous disc decompression using coblation technology (nucleoplasty) were prospectively enrolled. Patients were questioned preoperatively, postoperatively, and 24 hrs, 72 hrs, 1 wk, and 2 wks postprocedure by an independent reviewer regarding 17 possible symptom complications, which included bowel or bladder symptoms, muscle spasm, new pain, numbness/tingling or weakness, fevers/chills, rash/pruritus, headaches, nausea/vomiting, bleeding, and needle insertion site soreness. Statistical analysis was performed using Wilcoxon's signed-rank test. RESULTS: A total of 53 patients enrolled, of whom four patients dropped out. Two patients had increased symptoms and opted for surgery. Two patients could not be contacted. The most common side effects at 24 hrs postprocedure was soreness at the needle insertion site (76%), new numbness and tingling (26%), increased intensity of preprocedure back pain (15%), and new areas of back pain (15%). At 2 wks, no patient had soreness at the needle insertion site or new areas of back pain; however, new numbness and tingling was present in 15% of patients. Two patients (4%) had increased intensity of preprocedure back pain. There were statistically significant reductions in visual analog scale score for back pain and leg pain (P < 0.05). CONCLUSIONS: Based on this preliminary data, nucleoplasty seems to be associated with short-term increased pain at the needle insertion site and increased preprocedure back pain and tingling numbness but without other side effects. PMID- 16357544 TI - Adverse effects of fluoroscopically guided interlaminar thoracic epidural steroid injections. AB - OBJECTIVES: To assess the prevalence of adverse effects or complications from fluoroscopically guided thoracic interlaminar epidural steroid injections. DESIGN: A retrospective study with independent observer review. Patients presenting with thoracic radicular pain, caused by either herniated nucleus pulposus or thoracic spondylosis as confirmed by magnetic resonance imaging, received an interlaminar thoracic epidural steroid injection as part of a conservative-care treatment plan. The study was performed in a multidisciplinary spine care center. All injections were performed over a 5-yr period. An independent observer reviewed medical charts, which included a 24-hr postprocedure standardized questionnaire completed by telephone by an ambulatory surgical center nurse. Ambulatory surgical center operative reports and physician follow up office notes up to 3 mos after the procedures, along with epidurograms, were also reviewed. RESULTS: A total of 21 patients who received 39 injections were reviewed. Adverse effects or complications per injection observed included three with increased pain at injection site (7.7%), two with facial flushing (5.1%), one transient nonpositional headache (2.6%), one episode of insomnia the night of the injection (2.6%), and one episode of fever the night of the procedure (2.6%). Statistical analysis revealed no significant difference based on diagnosis (herniated nucleus pulposus vs. spondylosis, P = 0.9156), and age was not linked to higher prevalence of adverse/effects complications (P = 0.3137). CONCLUSIONS: No major complication arose. Adverse effects did occur with a rate of 20.5%. All adverse effects resolved without morbidity. No statistical difference was observed in the rate of adverse effects in patients with herniated nucleus pulposus or spondylosis. PMID- 16357545 TI - Breathing intolerance index: a new indicator for ventilator use. AB - OBJECTIVES: The purpose was to study the efficacy of a proposed breathing intolerance index for justifying ventilator use by patients with pulmonary or chest wall diseases and to compare with values obtained from healthy controls. DESIGN: A total of 42 patients with lung/chest wall disease, including 11 ventilator users and 25 age-matched controls, were studied. The breathing intolerance index was defined as (Ti/Ttot) x (Vt/VC), where Ti = inspiratory time of one breath (in seconds), Ttot = total time of one breath (in seconds), Vt = tidal volume (in milliliters) at rest, and VC = vital capacity (in milliliters). A digital spirometer with custom computer software was used. RESULTS: The examinations were completed uneventfully. The mean breathing intolerance index values of the 25 healthy volunteers, the 31 nonventilator user patients, and the 11 users of nocturnal noninvasive positive-pressure ventilation were 0.050 +/- 0.009 (mean +/- standard deviation), 0.087 +/- 0.022, and 0.186 +/- 0.038, respectively. The breathing intolerance indices of the ventilator users were significantly greater (P < 0.0001) than those of the other nonventilator user groups, and there was no overlap in values. CONCLUSIONS: Ventilator users have a significantly higher breathing intolerance index than nonventilator users. The index may be useful for justifying ventilator use. PMID- 16357546 TI - Ultrasound-guided shoulder injections in the treatment of subacromial bursitis. AB - OBJECTIVE: To investigate the treatment effectiveness between ultrasound-guided and blind injection techniques in the treatment of subacromial bursitis. DESIGN: A total of 40 patients with sonographic confirmation of subacromial bursitis were recruited into this study. These patients were divided into blind and ultrasound guided injection groups. The shoulder abduction range of motion was compared before injections and 1 wk after the completion of injections in both groups. RESULTS: The shoulder abduction range of motion before injection in the blind injection group was 71.03 +/- 12.38 degrees and improved to 100 +/- 18.18 degrees 1 wk after the injection treatments. However, the improvement did not reveal significant statistical differences (P > 0.05). The shoulder abduction range of motion before injection in the ultrasound-guided injection group was 69.05 +/- 14.72 degrees and improved to 139.29 +/- 20.14 degrees 1 wk after the injection treatments (P < 0.05). CONCLUSIONS: Ultrasound may be used as an adjuvant tool in guiding the needle accurately into the inflamed subacromial bursa. The ultrasound guided injection technique can result in significant improvement in shoulder abduction range of motion as compared with the blind injection technique in treating patients with subacromial bursitis. PMID- 16357547 TI - Steroid injections in the management of trigger fingers. AB - OBJECTIVES: The most commonly used primary treatment for trigger fingers is corticosteroid injection in the flexor tendon sheath, followed by surgical release if unsuccessful. This study examines the surgical and nonsurgical treatment of patients with trigger fingers presenting to a large Canadian tertiary referral center. The treatment success and side-effect profile of steroid injection therapy and surgical release were examined in the context of comorbid illness, specifically, diabetes mellitus. DESIGN: Retrospective review of all patients with trigger finger who were seen by the senior authors between January 1999 and June 2004. RESULTS: In the study period, 118 trigger digits were treated. This study included 92 nondiabetic, 21 type 2 diabetic, and five type 1 diabetic trigger fingers. Of the 89 digits that received at least one steroid injection, 46 (52%) resolved completely and 42 (47%) were improved. Nondiabetic digits were treated successfully in 40 out of 70 digits (57%) with steroid injection therapy. Diabetic patients had a success rate of 6 of 19 (32%) with steroid injections, which is significantly lower than nondiabetics (P = 0.04). All type 1 diabetics (n = 5) required surgical treatment. Surgical treatment was successful in 71 of 72 (99%) digits. No side effects of steroid injection were noted, and short-term postoperative side effects were noted in 26 of 72 surgical patients (36%). No statistically significant differences were found in surgical complication rates in diabetics vs. nondiabetics or type 1 diabetics vs. type 2 diabetics. CONCLUSIONS: Steroid injection therapy should be the first-line treatment of trigger fingers in nondiabetic patients. In diabetics, the success rate of steroid injection is significantly lower. Injection therapy for type 1 diabetics was ineffective in this study. Surgical release of the first annular (A1) pulley is most effective overall in diabetics and nondiabetics alike, with no higher rates of surgical complications in diabetics. PMID- 16357548 TI - Predictors of a successful driver evaluation in stroke patients after discharge based on an acute rehabilitation hospital evaluation. AB - OBJECTIVE: One of the most common concerns of a stroke patient is the ability to drive. We aimed to determine which neurologic impairments on an acute rehabilitation admission evaluation predict the likelihood of a successful driver evaluation after discharge. DESIGN: Prospective study in an acute stroke rehabilitation unit. RESULTS: A total of 45 stroke patients undertook a driver evaluation at our institution. The mean age +/- standard deviation was 71.0 +/- 9.8 yrs, Mini-Mental State Examination score was 22.7 +/- 8.1, upper limb and lower limb Motricity Index scores were 63.7 +/- 34.8 and 71.8 +/- 24.3, Limb Placement Task was 4.6 +/- 3.6 inches, and admission total FIM score was 68.5 +/- 18. The admission variables differed between those who failed (n = 10) vs. those who passed the in-clinic driver evaluation (n = 29, 75%): Mini-Mental State Examination (17.5 +/- 9.7 vs. 24.6 +/- 6.7, P = 0.004), and upper limb (82 +/- 23.7 vs. 57.4 +/- 36.1, P = 0.05) and lower limb (87.6 +/- 11.8 vs. 66.4 +/- 25.2, P = 0.01) Motricity Index scores. CONCLUSIONS: Patients who undertook and passed the in-clinic driver evaluation had, at admission, higher Mini-Mental State Examination and Motricity Index scores with normal visual field defects. PMID- 16357549 TI - Shoulder impingement syndrome: relationships between clinical, functional, and radiologic findings. AB - OBJECTIVE: Although there has been much research about imaging methods for shoulder impingement syndrome, the clinical information and upper limb level of disability have been generally ignored. The purpose of this study was to detect the relationships between clinical, functional, and radiologic variables in patients with shoulder impingement syndrome. DESIGN: A cross-sectional, clinical, and radiologic study was planned and 59 shoulders of 58 consecutive patients waiting for physical therapy because of a clinically suspected shoulder impingement syndrome were included into this study. Comprehensive clinical examination, radiography, shoulder ultrasonography, and magnetic resonance imaging were performed in the same month. RESULTS: Despite the high sensitivities of ultrasonography for diagnosing rotator cuff tears (98.1%) and biceps pathologies (100%), magnetic resonance imaging was superior to ultrasonography in many important shoulder structures such as a glenoid labral tear and subacromial bursal effusion/hypertrophy (P < 0.01). These structures were the determinants of the shoulder's disability measured by disabilities of the arm, shoulder, and hand questionnaire. CONCLUSION: Ultrasonography and magnetic resonance imaging had comparable high accuracy for identifying the biceps pathologies and rotator cuff tears. The basic clinical tests had modest accuracy in both disorders. The choice of which imaging test to perform should be based on the patient's clinical information (regarding lesion of glenoid labrum, joint capsule, muscle, and bone), cost, and imaging experience of the radiology department. PMID- 16357550 TI - Functional and neuromotor performance in older adults: effect of 12 wks of aerobic exercise. AB - OBJECTIVE: There is little information regarding the effect of aerobic exercise on physical and neuromotor performance in older adults. The aim of this randomized, controlled trial was to determine the effects of a 12-wk aerobic exercise program on functional and neuromotor performance in inactive healthy older adults. DESIGN: A total of 22 inactive healthy older adults, aged 60-75 yrs, voluntarily participated in the study and were randomly assigned to a progressively increased high-intensity aerobic exercise group (n = 12) or to a control group (n = 10). The aerobic exercise group exercised three times per week for 12 wks on nonconsecutive days. Both groups were evaluated before and after the exercise period in the one-repetition maximum of knee extensors and flexors, 6-min walk distance, chair-rising time, and whole-body reaction time. RESULTS: Multivariate analysis of variance repeated measures (2 x 2) revealed significant (P < 0.05) time by group interaction for one-repetition maximum knee extension and flexion strength, 6-min walk distance, chair-rising time, and whole-body reaction time. For the aerobic exercise group, the following variables improved significantly (P < 0.05) after the exercise period: the one-repetition maximum knee extensors (12%) and flexors (19%) muscle strength, 6-min walk distance (17%), chair-rising time (8%), and whole-body reaction time (20%). No significant differences were observed in the control group. CONCLUSIONS: The results of the present study show that short-term progressively increased high-intensity aerobic exercise improved the physical and neuromotor performance in inactive healthy older adults. These findings suggest that the participation in a progressively increased high-intensity aerobic exercise program may improve mobility and ability to carry out activities of daily living in older adults. PMID- 16357551 TI - Ambulation training with and without partial weightbearing after traumatic brain injury: results of a randomized, controlled trial. AB - OBJECTIVE: To test the hypothesis that 8 wks of partial weight-bearing gait retraining improves functional ambulation to a greater extent than traditional physical therapy in individuals after traumatic brain injury. DESIGN: A randomized, open-label, controlled, cohort study was conducted at two inpatient university-based rehabilitation hospitals. A total of 38 adults with a primary diagnosis of traumatic brain injury and significant gait abnormalities received either 8 wks of standard physical therapy or physical therapy supplemented with partial weight-bearing gait training twice weekly. RESULTS: Significant (P < 0.05) improvements were detected in both groups on Functional Ambulation Category, Standing Balance Scale, Rivermead Mobility Index, and FIM. However, no differences were found between the treatment groups. CONCLUSIONS: Results did not support the hypothesis that 8 wks of partial weight-bearing gait retraining improves functional ambulation to a greater extent than traditional physical therapy in individuals after traumatic brain injury based on common clinical measures. PMID- 16357552 TI - Low-dose botulinum toxin with short-term electrical stimulation in poststroke spastic drop foot: a preliminary study. AB - OBJECTIVE: To compare low-dose botulinum toxin injections followed by short-term electrical stimulation with high-dose botulinum toxin for poststroke spastic drop foot. DESIGN: Twelve patients with spastic drop foot were randomly assigned to two treatment groups. The first group (n = 6) received low-dose (100 units) botulinum toxin injection to the posterior tibial muscle in combination with short-term electrical stimulation, and the second group (n = 6) received a total of 400 units of botulinum toxin injections in equal doses to the posterior tibial, soleus, medial, and lateral gastrocnemius muscles. The patients were evaluated before and 2, 4, 8, and 12 wks after the treatment by the following variables: resting position angle, active and passive ankle range of motion, Modified Ashworth Scale, time walking 10 m, clonus score, Brace Wear Scale, and Global Assessment of Spasticity Scale. RESULTS: Improvement was recorded in both groups for all variables except brace wear. No significant difference was found between the study groups after treatment. The effects lasted shorter in the first group, whereas more side effects were seen in the second group. CONCLUSIONS: Our preliminary single-blind study was unable to find a difference in terms of effectiveness between low-dose (100 units) botulinum toxin combined with short term electrical stimulation and high-dose (400 units) application in spastic drop foot. PMID- 16357553 TI - Selective tibial neurotomy in the treatment of spastic equinovarus foot: a 2-year follow-up of three cases. AB - OBJECTIVE: To objectively assess the decrease in spasticity and the improvement in gait after tibial nerve neurotomy performed to treat spastic equinovarus foot. DESIGN: Before-after trial with a 2-yr follow-up. Three hemiplegic patients with spastic equinovarus foot were treated with a selective peripheral neurotomy of the tibial motor nerve branches (soleus, lateral and medial gastrocnemius and tibialis posterior nerves). Evaluation included clinical assessment of spasticity (Ashworth scale), maximal Hoffmann reflex (H(max))/compound muscle action potential (M(max)) ratio measurement, gait analysis, and muscle stiffness evaluation performed before and 2 mos, 1 yr, and 2 yrs after the neurotomy. RESULTS: Spasticity, muscle stiffness, and H(max)/M(max) ratio decreased after neurotomy. The kinematic (ankle dorsal flexion and knee recurvatum) and kinetic variables (maximum ankle muscle moment and external work) of the gait were permanently improved after neurotomy. Interestingly, kinetic variables seemed to gradually improve with time after the neurotomy. CONCLUSION: Tibial neurotomy is an effective and durable treatment for spastic equinovarus foot. PMID- 16357554 TI - Effectiveness of upper and lower limb casting and orthoses in children with cerebral palsy: an overview of review articles. AB - The objective of this overview is to summarize from systematic reviews the evidence on the effectiveness of using upper and lower limb casting or orthoses in children with cerebral palsy. We used computerized bibliographic databases to search for systematic reviews without any language restrictions. Identification, selection, quality assessment, and data extraction were performed independently by two investigators. Of the 40 identified reviews, 23 were selected for closer consideration, and five reviews met the inclusion criteria. The quality of existing systematic reviews and original studies included in our reviews varied widely. The following evidence was found: (1) casting of lower limbs has a short term effect on passive range of movement; (2) orthoses that restrict ankle plantar flexion have a favorable effect on an equinus walk, but the long-term clinical significance is unclear; (3) evidence on managing upper limb problems with casting or splinting in children with cerebral palsy is inconclusive. Our conclusion is that there is a paucity of evidence from primary studies on the use of orthoses in children with cerebral palsy. More original, well-designed research is needed. PMID- 16357555 TI - Long cervical costae articulating with the first ribs: a salient case of thoracic outlet syndrome. PMID- 16357557 TI - Evolving concepts in pheochromocytoma and paraganglioma. AB - PURPOSE OF REVIEW: The pheochromocytoma field has recently undergone a paradigm shift. This review will highlight some of these novel findings, including their impact on our understanding of the disease biology and influence on clinical management. RECENT FINDINGS: Identification of novel susceptibility loci and recognition of a high rate of germline mutations in pheochromocytomas indicate that their genetic diversity is broader and more complex than previously estimated. Further, increased risk of tumor malignancy and aggressiveness in certain patients with succinate dehydrogenase subunit B(SDHB) mutations suggest that they may have prognostic value as predictors of pheochromocytoma behavior. Finally, discovery of a shared activation of the hypoxic response in pheochromocytomas with mutations in VHL and SDH genes and uncovering of a common JunB-mediated apoptosis defect in the major hereditary groups of pheochromocytoma have provided a mechanistic basis for the clinical similarities between these distinct syndromes. SUMMARY: The notion that 'sporadic'-appearing tumors may in fact be components of one of multiple hereditary syndromes has a major impact on surveillance and follow-up of patients and their at-risk family members. Likewise, the ability to predict tumor malignancy has the potential to improve the prognosis of these patients. Importantly, insights into the biology of pheochromocytomas have provided clues on pathway interactions in cancers and have laid the ground for generation of new hypotheses on the cell-of-origin of these tumors. Pheochromocytomas have therefore emerged as key models for understanding cancer biology and for paving the way for future designer treatment in this and other cancers. PMID- 16357558 TI - Treatment of advanced carcinoid tumors. AB - PURPOSE OF REVIEW: Carcinoid tumors often present with metastatic disease. Generally, these tumors can be treated conservatively. New evidence exists, however, that stage IV disease may be better managed with more aggressive medical and surgical treatment. Headway is also being made into understanding the associated fibrosis seen with advanced disease and in better understanding signaling pathways with the hope of offering future treatment options. RECENT FINDINGS: Recent literature has advocated for more aggressive surgical treatment of carcinoid tumors, especially in the setting of hepatic metastases and peritoneal carcinomatosis. Octreotide and lanreotide are further being described for treating metastatic carcinoids. Radiolabeled somatostatin analogues may prove to be as effective for treating carcinoids as for visualizing them. Other potential treatment modalities include pharmacologic activation of signaling pathways to control excess hormone production. Research into fibrosis - a cause of pain, bowel obstruction, retroperitoneal vascular constriction and right heart failure - has shown that serotonin and tachykinins may be the key mediators. SUMMARY: Patients with stage IV carcinoid tumors may benefit from more aggressive surgical management and new treatment modalities. The growing body of knowledge regarding important molecular signaling pathway may lead to new medical therapies and further understanding of the sequelae of excess hormone production. PMID- 16357559 TI - Parathyroid carcinoma. AB - PURPOSE OF REVIEW: This article reviews current knowledge on the etiology, diagnosis and treatment of parathyroid carcinoma. RECENT FINDINGS: Due to its rarity, research on the molecular etiology and treatment of parathyroid carcinoma has been slow. Mutations of the tumor suppressor gene, HRPT2, and resultant loss of expression of its gene product have been found in the majority of parathyroid cancers studied. Recent advances in the field have identified regions on several chromosomes that demonstrate loss of heterozygosity more commonly in parathyroid carcinoma than in benign parathyroid lesions. This has provided clues to the location and identity of additional tumor suppressor genes associated with the development of this cancer. SUMMARY: Parathyroid carcinoma is an extremely rare cause of primary hyperparathyroidism, accounting for fewer than 1% of cases. The etiology of parathyroid cancer is largely unknown. Associations have been made with several inherited syndromes and with specific genetic lesions. Little is known about the most appropriate management of this disease. En bloc resection at the time of initial surgery appears to provide the best chance of cure. Anecdotal experience with adjuvant chemotherapy has shown a modest and short-lived effect. External beam radiation following surgical resection, however, may increase long term survival compared with surgery alone. Bisphosphonates and a new class of drugs known as calcimimetics have been used effectively in some patients to control the symptoms of severe hypercalcemia in a palliative setting. PMID- 16357560 TI - Endocrine tumors of the pancreas. AB - PURPOSE OF REVIEW: Neoplasms of the endocrine pancreas, commonly referenced as pancreatic islet cell tumors, are rare, often well differentiated endocrine neoplasms, whose biology remains poorly characterized. This article reviews the current clinical management of pancreatic islet cell tumors and describes the molecular events that have been studied to guide future therapies of these peculiar neoplasms. RECENT FINDINGS: While some islet cell tumors arise in association with the MEN-1 syndrome, the majority of these neoplasms are sporadic lesions whose underlying genetic and molecular events remain largely unknown. Recent work has identified changes in gene expression occurring in metastatic and non-metastatic islet cell tumors, which appear to correlate with the occurrence of lymph node and liver metastases. Epigenetic alterations of select tumor suppressor genes may influence patient survival, and the presence of gene promoter methylation may be used as a prognostic marker system. In addition, multiple molecular alterations, including changes in expression of cellular proteins with migratory, cell cycle or angiogenic functions, have been demonstrated to influence islet cell tumor growth, invasion and metastatic spread. SUMMARY: Understanding the molecular events underlying the biology of pancreatic islet cell tumors will aid the development of accurate prognostic markers and will guide improved therapeutic modalities in the future. PMID- 16357561 TI - Thyroid carcinoma. AB - PURPOSE OF REVIEW: Diagnostic methods and treatment options for differentiated and medullary thyroid carcinoma are continuously influenced by new trends and techniques. Our review therefore displays the most recent clinical practices for diagnosis and operative treatment of differentiated and medullary thyroid carcinoma. RECENT FINDINGS: Among the new diagnostic methods, high-resolution ultrasonography plays an important role for both the evaluation of thyroid nodules and the detection of enlarged suspicious cervical lymph nodes. The results of ultrasound will definitely influence operative decisions. A second step to diagnose medullary thyroid carcinoma is calcitonin measurement before surgery, which is sensitive and specific enough to detect medullary thyroid carcinoma in patients with thyroid nodules. Surgical treatment for differentiated or medullary thyroid carcinoma mostly consists of total or near-total thyroidectomy. An additional central or lateral modified-radical neck dissection might help to reduce local recurrence, especially in medullary carcinoma, but still does not influence significantly the survival rates. Monitoring of the recurrent laryngeal nerve during surgery is used increasingly. According to the newest literature, however, compared with visual identification of the laryngeal nerve, it cannot be considered as superior. SUMMARY: Diagnosis and treatment of thyroid carcinoma are still subjected to changes and the different options of surgical treatment in particular will be continuously discussed in the future. PMID- 16357562 TI - Adrenocortical carcinoma. AB - PURPOSE OF REVIEW: Adrenocortical carcinoma is a rare malignancy, accounting for 0.02% of all annual cancers reported. Given the generally advanced stage at diagnosis, the overall 5-year survival remains poor, varying between 20 and 45%. While older studies purported an improved outcome for functional tumors in adult patients, this has not been borne out in more recent studies. In the pediatric population, though, virilizing tumors carry a better survival than non-functional or cortisol-secreting tumors. RECENT FINDINGS: Recent studies focusing on the tumorigenesis of adrenocortical carcinoma have focused on onco-developmental genes present in the fetal adrenal cortex, as well as local adrenal paracrine and autocrine effects of cellular peptides. SUMMARY: Pre-operative diagnostic advances in positron emission scanning are emerging as promising modalities for confirmation of malignancy of indeterminate adrenal masses. No significant advances in the treatment of adrenocortical carcinoma have been developed. Surgery remains the mainstay for primary and recurrent disease, including select patients with isolated liver metastases. Mitotane has remained the preferred adjuvant treatment agent, showing modest effect in patients with unresectable, residual or metastatic disease. Multi-institutional registries and trials need to be established, with multidisciplinary efforts focused on the development of new therapeutic strategies. PMID- 16357563 TI - Minimally invasive thyroidectomy. AB - PURPOSE OF REVIEW: Based on recent advances in ultrasonography, color-Doppler sonography and ultrasound-guided fine needle aspiration biopsy, minimally invasive thyroidectomy has been widely employed for treating thyroid tumors. Minimally invasive procedures have recently been adopted for the surgical treatment of thyroid disease as a means of preventing prominent scars, hypesthesia and paresthesia. RECENT FINDINGS: We have developed axillary and anterior chest approaches for endoscopic thyroidectomy. The mean operating time for the endoscopic procedure was significantly longer than that for conventional open surgery. All patients who were treated using the axillary approach were satisfied with the cosmetic results. The incidence of complaints after endoscopic surgery is significantly lower than that after open surgery. The minimally invasive mini-incision approach has been performed under direct and endoscopic vision to minimize surgical invasiveness and improve cosmetic results. Our procedure differs from conventional thyroidectomy in requiring a 3-cm skin incision, and in not raising of a skin flap and not dissecting the sternohyoid muscle. SUMMARY: Endoscopic thyroidectomy is the procedure of choice in well selected patients with thyroid disease. The present indications for this procedure include benign follicular adenomas less than 4 cm in diameter, low-risk papillary carcinomas less than 10 mm, oxyphilic cell tumors less than 4 cm and Graves' disease as imaged on preoperative ultrasonography. The minimally invasive mini-incision approach is indicated for patients with low-risk, well differentiated carcinomas. Total thyroidectomy and bilateral modified neck dissection is possible, if the tumor is not large. PMID- 16357564 TI - Tumor cell detection in peripheral blood and bone marrow. AB - PURPOSE OF REVIEW: Whether the occurrence of tumor cells in peripheral blood or bone marrow from patients with solid tumors is predictive for disease recurrence or of any other prognostic relevance remains unknown. This article reviews recently published results focusing on the various methods used, their correlations with clinical or biological parameters and their potential prognostic value. RECENT FINDINGS: An increasing number of marker genes and different techniques, alone or in combinations, have been used for the detection of tumor cells in peripheral blood and bone marrow. Various results obtained are hardly comparable, most often due to the different methods in use. The frequency of circulating tumor cells in peripheral blood varied within a broad range and their clinical relevance appeared to be contradictory, at least in part. Disseminated tumor cells in bone marrow reached an independent prognostic value in breast cancer patients, but several investigations led to inconsistent correlations with clinical or prognostic criteria. SUMMARY: Still many questions remain unanswered; hence, the detection of tumor cells in peripheral blood or bone marrow cannot yet be taken into account for therapeutic decisions. PMID- 16357565 TI - Gene expression determinants of clinical outcome. AB - PURPOSE OF REVIEW: Gene expression profiling of human cancers has produced a large amount of information that could aid physicians in making decisions related to the treatment of their patients. A number of recent studies have identified collections of genes and corresponding expression levels that correlate with the physical appearance and clinical behavior of all common human neoplasms. RECENT FINDINGS: This article will discuss emerging trends of study design and approaches to gene expression signature identification. Recently discovered gene expression signatures that correlate with cellular morphology, resistance to chemotherapy, and overall clinical course of disease are discussed. SUMMARY: The prospective evaluation of the predictive value of a number of gene signatures is now appropriate. Newly discovered gene expression signatures that correlate with disease clinical course should be independently evaluated. The timely reporting of both positive and negative results from such validation attempts will be crucial to the advancement of molecular pathology. PMID- 16357566 TI - Chromosomal translocations in cancer and their relevance for therapy. AB - PURPOSE OF REVIEW: Recurring chromosomal abnormalities are considered the primary genetic change in oncogenesis as well as an important indicator for tumor phenotype and clinical outcome. This review highlights recent findings regarding the genes associated with chromosomal translocations. RECENT FINDINGS: A great number of novel fusion genes associated with chromosomal translocations have been cloned. These novel fusion genes are found in the smaller part of various malignancies, and it can be expected that the significance of novel fusion gene occurrence for oncogenesis will be clarified in the not too distant future. Observation of high frequencies of mutations in NOTCH1, NPM and JAK2 in T-cell acute lymphoblastic leukemia, acute myeloid leukemia with normal karyotype and myeloproliferative disorders (polycythemia vera, essential thrombocythemia and idiopathic myelofibrosis) have provided important suggestions for a better understanding of chromosomal translocations. This is because all these genes had already been identified as genes associated with chromosomal translocations in a small subset of specific phenotypes of hematologic malignancies. SUMMARY: This review summarizes recent findings associated with chromosomal translocations including newly identified fusion genes, a novel mechanism of fusion gene formation and their relevance for novel targeted therapies. Continuing attempts to identify genes associated with chromosomal translocations can be expected to provide further insights into the significance of various gene alterations in cancer and the development of novel targeted therapies. PMID- 16357567 TI - Kinase mutations in cancer: chinks in the enemy's armour? AB - PURPOSE OF REVIEW: Over the past few years, a revolution has transformed the oncology field. This revolution is characterized by two main features. The first is the introduction of the concept of individualized cancer therapy. The second is the development of drugs targeting molecules selectively altered in tumours. This review analyses these aspects by looking at the role that altered kinases and their inhibitors have played in this historical process. RECENT FINDINGS: Tumour progression is the result of the sequential accumulation of mutations in genes monitoring the rates of cell birth and cell death. The molecular profiling of cancers has shown that protein and lipid kinases are frequently altered in tumour cells. In most cases, these alterations translate in constitutively active proteins, which are amenable of therapeutic targeting. Intriguingly, even 'established' cancer cells remain somewhat 'addicted' to the deregulated activity of mutated kinases. This feature appears to be the basis for the ability of kinase inhibitors in controlling the development of a number of cancers. The therapeutic efficacy of kinase inhibitors is impaired by the emergence of tumour cells carrying 'resistance' mutations. SUMMARY: Many oncogenes are mutated kinase genes. In most cases, the mutations result in the constitutive activation of the affected kinase that can be pharmacologically inhibited. Unfortunately, upon treatment with kinase inhibitors, resistant clones develop rapidly, impairing their therapeutic effect. Strategies to overcome resistance are discussed as well as the possibility to target kinases regulating cancer stem cells. PMID- 16357568 TI - Oncogenic PI3K and its role in cancer. AB - PURPOSE OF REVIEW: The purpose of this review is to examine the contribution of the PI3K signaling pathway to the development of human tumors and to propose further studies to elucidate how to develop therapeutics for patients with mutations in this pathway. RECENT FINDINGS: More than 30% of various solid tumor types were recently found to contain mutations in PIK3CA, the catalytic subunit of PI3K. Further analysis of key genes in this pathway identified an additional eight genes altered in tumors. These were generally found to be mutated in a mutually exclusive manner, thus increasing the mutation frequency of the pathway to 40% in colorectal cancers and emphasizing the importance of the PI3K pathway in tumorigenesis. Functional analyses of PIK3CA mutations revealed that they increase its enzymatic activity, stimulate AKT signaling, allow growth factor independent growth as well as increasing cell invasion and metastasis. SUMMARY: The PI3K signaling pathway is dysregulated by a variety of mechanisms in a large fraction of human tumors. Both mutational and functional analyses have shown that PIK3CA is an oncogene that plays an important role in tumor progression. Mutant members of the PI3K pathway, including PIK3CA, are good targets for therapeutic intervention because most of them are kinases, making them attractive for drug development. Gaining further insights into PIK3CA oncogenic mechanisms may produce new biomarkers and help the development of targeted therapeutics. PMID- 16357569 TI - Bibliography. Current world literature. Endocrine tumors. PMID- 16357571 TI - Mucus and MUC in asthma. AB - PURPOSE OF REVIEW: Asthma is characterized by chronic airway inflammation and a mucus hypersecretory phenotype comprising excess mucus secretion, goblet cell hyperplasia and submucosal gland hypertrophy. This augmented mucus secretion has been relatively undervalued in asthma compared with airway inflammation. However, mucus plugging contributes to airflow limitation and airway hyperresponsiveness, and to morbidity and mortality in asthma. We review recent contributions to this field and therapeutic avenues to control mucus hypersecretion. RECENT FINDINGS: A distinct mucus hypersecretory phenotype may present in asthma. Overexpression of MUC5AC, MUC5B and MUC2 have been described in asthma secretions, but identification of defined biochemical abnormalities and polymorphisms of mucin genes linked to asthma remains elusive. Activation of epidermal growth factor receptor (EGFR) activation appears central in transducing many different stimuli, including oxidative stress, proteases and cytokines. In contrast, nitrosative stress has barely been investigated. The existence of crosstalk between EGFR and other receptor systems may provide new clues regarding the activity of acetylcholine, adenosine and other agonists of G-protein-coupled receptors and other receptor families on mucin secretion. Modern techniques for noninvasive detection of mucus pathology will advance clinical research in this field. SUMMARY: Airway mucus hypersecretion as a part of airway remodelling represents a problem in asthma, and studies of pathophysiology and therapeutic approaches are therefore warranted. Identification of targets such as the EGFR cascade, which are crucial in excessive and abnormal mucus secretion, may lead to the rational design of new antihypersecretory drugs that may enhance future asthma treatment. PMID- 16357572 TI - Reactive oxidant species in asthma. AB - PURPOSE OF REVIEW: This overview summarizes some recent studies on the balance of oxidants to antioxidants in patients with asthma. The aim of the review is to compare studies on the changes in oxidants/antioxidants in stable asthma or in acute exacerbation of asthma. RECENT FINDINGS: Our review of the recent literature in this field seems to indicate conflicting findings. Increased release of reactive oxygen species such as superoxide anion and hydrogen peroxide has been reported in exhaled breath condensates and from circulating granulocytes, and from the bronchoalveolar lavage cells of patients with asthma. In asthma, bronchial obstruction is associated with an increased spontaneous and stimulus-induced production of oxygen free radicals. The primary defense against reactive oxygen species is endogenous antioxidants, which are found to be altered in asthma. A marked decrease in plasma antioxidant capacity occurs. Superoxide dismutase activity is higher in erythrocytes and serum of asthmatic than in normal subjects and is diminished in cells from lavage and brushing samples of patients with asthma. Higher level of erythrocyte catalase activity has only been found in Chinese asthmatic patients while decreased glutathione peroxidase activity has been well documented. SUMMARY: Since there are considerable discrepancies in erythrocyte or plasma antioxidant enzyme activity in patients with asthma, the problem at this time is attempting to sort out these conflicting results and to find their roles in the pathogenesis of asthma. There is good evidence that antioxidant compounds may have a potential role in the treatment of asthma, especially of asthma exacerbation. PMID- 16357573 TI - The significance of beta2-adrenergic receptor polymorphisms in asthma. AB - PURPOSE OF REVIEW: The gene that encodes the beta2-adrenergic receptor (ADRB2) is one of the most studied candidate genes in asthma. This review examines recent articles of the relationship between ADRB2 polymorphisms and asthma. RECENT FINDINGS: Candidate gene association studies of ADRB2 and asthma have been dominated by analyses of the two common non-synonymous coding single nucleotide polymorphisms, Arg16Gly and Glu27Gly. Published studies have yielded inconsistent results. Three recent meta-analyses on the effects of these two polymorphisms have found no associations with asthma, although there were suggestions of associations with other asthma-related phenotypes, such as nocturnal asthma and asthma severity. Other recent studies have investigated other single nucleotide polymorphisms in this gene (i.e. single nucleotide polymorphisms in the promoter region and other single nucleotide polymorphisms in the coding region). These analyses have investigated the association between these single nucleotide polymorphisms (and haplotypes of these polymorphisms) and asthma-related phenotypes such as lung function, airways hyperresponsiveness, and response to a bronchodilator, and have suggested that certain regions of the gene may be associated with different phenotypes. Results from these studies, however, have also been inconsistent. SUMMARY: Polymorphisms of ADRB2 are not major risk factors for the development of asthma. These polymorphisms are likely to be important, however, in determining drug response. Future studies need to fully characterize all of the variation in the gene and perform comprehensive association studies. Finally, interactions between ADRB2 and other genes in the beta-agonist pathway are an important and active area of research that will shed more light on inter-individual differences in drug response. PMID- 16357574 TI - Dyspnea and asthma. AB - PURPOSE OF REVIEW: Dyspnea--the perception of respiratory discomfort--is a primary symptom of asthma. This review examines possible ways to link mechanisms, measurement and treatment that will increase our understanding of this condition. RECENT FINDINGS: Functional neuroimaging methods have proven to be powerful tools that serve as advanced models of sensor motor brain function. Studies examining functional neuroimaging methods have revealed activation of distinct brain areas associated with increased dyspnea. Pulmonary hyperinflation has been proposed to influence the perception of dyspnea. The association of hyperinflation with minor levels of bronchoconstriction reflects the partition of the sensory effect of airway narrowing per se from that of the attendant elastic loading of the inspiratory muscles. There is evidence to suggest, however, that hyperinflation does not play an important role in the pathogenesis of exercise dyspnea as it does during induced bronchoconstriction. Decreased levels of perception of airway obstruction may be a risk factor associated with life-threatening asthma. A poor perceiver may be vulnerable to further hypoxia-induced suppression of respiratory sensation. Monitoring the response to bronchodilator therapy with formoterol and salbutamol in patients with acute or chronic asthma has resulted in significantly faster improvement in dyspnea, within 2 min. SUMMARY: Regardless of the factors involved, much variability in dyspnea scores remains unexplained. Quantitative and qualitative assessment of the perception of dyspnea, symptom measurement and quality of life complement physiological measurements and contribute to our understanding of dyspnea in asthma. PMID- 16357575 TI - Asthma screening, case identification and treatment in school-based programs. AB - PURPOSE OF REVIEW: Asthma is an important health problem in school-aged children and schools seem an obvious site to find and work with under-recognized and under treated asthma. Teachers and coaches often must deal with asthma-related symptoms or emergencies requiring knowledge, skills, and written plans and policies. In 2005, school-based asthma work focused on two areas: identification of unrecognized asthma and management of under-treated asthma. RECENT FINDINGS: Effective school-based screening requires a simple, effective screening tool. Three new asthma screening tools continue to identify more false-positive than true positive cases of asthma. Public health experts question whether asthma even fits the usual criteria for 'screening' because it does not have an asymptomatic phase. 'Case-finding' is presented as a better use of resources, allowing schools to focus on children with asthma that has been diagnosed but remains symptomatic. No school-based program based on letters, reminders, or recommendations sent to parents or community physicians changed asthma care. Three reports describe programs designed to supplement usual asthma care by providing in-school interventions, but none appeared ready for implementation in all schools in the USA. A major barrier was the continuing lack of school nurses, who must have asthma-related education and medical support to provide school-based asthma management. SUMMARY: Schools continue to be a site for asthma interventions but few of the programs, even the most intensive, influence children's asthma-related health. Most programs require modifications and further evaluation, and all require careful assessment of the burden on schools. PMID- 16357576 TI - Spirometry in the diagnosis of asthma in children. AB - PURPOSE OF REVIEW: To review the diagnostic accuracy of lung function measurements made using spirometry for childhood asthma, recent guidelines for the measurement and interpretation of spirometric lung function tests and recent developments for diagnosing asthma. RECENT FINDINGS: Measurements of lung function and bronchial lability made using spirometry may not perform any better than other tests such as skin prick testing, or measurements of exhaled nitric oxide for diagnosing asthma. New guidelines are available. SUMMARY: Spirometry is a simple, robust and widely available tool for investigating lung function. There are published guidelines for making measurements and their interpretation. The place of spirometry in the diagnosis of asthma, however, needs clarification. The diagnostic profiles of measurements of bronchodilator responsiveness and bronchial hyperreactivity made using spirometry, although reasonable, are not perfect. In schoolchildren, they are no better than knowledge of aeroallergen sensitization when considering a diagnosis of asthma. PMID- 16357577 TI - Asthma therapy for children under 5 years of age. AB - PURPOSE OF REVIEW: The evidence for effectiveness of currently used asthma medication for wheeze in young children is reviewed. RECENT FINDINGS: The management of the infant and preschool child with wheezing is complicated by the uncertainty with respect to the aetiology. Difficulties in defining phenotypes and objective outcome parameters combined with the transient nature of symptoms which often resolve spontaneously have confounded many therapeutic studies. Recent studies on the effect of pharmacotherapy in wheezing infants have tried to define a more homogeneous phenotype as well as make a selection of patients that are likely to respond to the studied drug. In addition, these studies have used lung function parameters and nitric oxide as one of the outcome measurements. Studies on the nature of inflammation and the development of airway remodelling in infants and young children are done to further define phenotypes. SUMMARY: Currently, there are no evidence-based guidelines and not even consensus statements on the right approach in pharmacological treatment of wheezing in infants and preschool children. The main issue still is the difficulty in coming to a correct diagnosis. Further studies are needed on the nature and the diagnostics of phenotypes and on the effect of early intervention. PMID- 16357578 TI - Predicting adult asthma in childhood. AB - PURPOSE OF REVIEW: There still is no cure for asthma. Early identification of patients at risk for disease progression may lead to better treatment opportunities and hopefully better disease outcomes in adulthood. Recent literature on childhood risk factors associated with the outcome of asthma in adulthood is reviewed. RECENT FINDINGS: The role of sex and smoking on the outcome of childhood asthma is still uncertain. Better lung function in childhood is consistently associated with better prognosis in adulthood, whereas the severity of bronchial hyperresponsiveness is not a good predictor of asthma outcome. Childhood atopy is associated with the persistence of symptoms but not with lung function in adulthood. An important finding is that anti-inflammatory treatment can reduce the accelerated decline in forced expiratory volume in 1 s and airway remodelling, but there is considerable variety in treatment response between individuals, which possibly can be explained by genetic make-up. Although many studies have investigated the genetics of asthma susceptibility, longitudinal studies on genes associated with asthma outcome are scarce. SUMMARY: Despite many high-quality investigations, accurate prediction of adult asthma in childhood is still not feasible. Future studies should focus on associations between genetic background and asthma prognosis as an important area of research with great potential. PMID- 16357579 TI - Initial corticosteroid therapy for asthma. AB - PURPOSE OF REVIEW: This review examines the commencement of maintenance pharmacotherapy for asthma: inhaled corticosteroids alone or in combination with long-acting beta2 agonists. RECENT FINDINGS: A systematic review of randomized controlled trials has examined the starting dose of inhaled corticosteroids (high, moderate, low) and the dose regimen (step down versus constant) in asthma. There was no significant difference in key asthma outcomes for step down compared with a constant inhaled corticosteroid dose. There was no significant difference between high or moderate dose inhaled corticosteroid groups (n=11) for morning peak expiratory flow, symptoms and rescue medication use. There may be a benefit from high-dose inhaled corticosteroids for airway hyperresponsiveness. There was a significant improvement in peak expiratory flow and nocturnal symptoms in favour of a moderate inhaled corticosteroid dose compared with low-dose treatment. Long-acting beta2 agonists combined with inhaled corticosteroids as initial asthma therapy has been examined in a systematic review of nine randomized controlled trials. Inhaled corticosteroids combined with long-acting beta2 agonists led to significant improvements in forced expiratory volume in 1 s, morning peak expiratory flow, symptom score and symptom-free days but no difference in exacerbations requiring oral corticosteroids. A randomized controlled trial of patients with uncontrolled asthma found a benefit of escalating doses of salmeterol/fluticasone compared with fluticasone on asthma control. SUMMARY: Initial inhaled corticosteroid therapy should begin with a constant, moderate dose. Initial therapy with long-acting beta2 agonist and inhaled corticosteroids achieves superior improvement in symptoms and lung function, and at a quicker rate than inhaled corticosteroids alone. There is no benefit in terms of reduced exacerbations unless an escalating inhaled corticosteroid dose strategy is used. PMID- 16357580 TI - Adenosine signaling in asthma and chronic obstructive pulmonary disease. AB - PURPOSE OF REVIEW: The chronic lung diseases, asthma and chronic obstructive pulmonary disease, are pulmonary disorders in which persistent inflammation and alterations in lung structure contribute to a progressive loss of lung function. Although the exact type of inflammation and damage in each disease is distinct, they share the common feature that they are chronic in nature. Despite efforts, little is known about the cellular and molecular mechanisms that drive the chronicity of these two diseases. This review will summarize important findings regarding the role of adenosine, a signaling nucleoside implicated in the pathogenesis of these two disorders. RECENT FINDINGS: Aerosolized adenosine induces bronchoconstriction in patients with asthma and chronic obstructive pulmonary disease primarily through the release of mast cell mediators. In this setting it can not only be used to aid in diagnosis but also to monitor patient responses to steroid therapy. Adenosine levels are elevated in the lungs of asthma patients, indicating greater flux through adenosine receptor signaling pathways. In-vitro studies have shown adenosine to access pathways leading to the genesis of chronic inflammation via the release of proinflammatory cytokines and chemokines. Animal studies demonstrate that merely elevating adenosine levels in the mouse is sufficient to induce a pulmonary phenotype with features of asthma and chronic obstructive pulmonary disease. SUMMARY: Identifying mediators regulating the chronic nature of asthma and chronic obstructive pulmonary disease is critical towards advancements in treatment options. Adenosine has been implicated in promoting the inflammation and airway remodeling seen in chronic lung disease and thus makes an attractive therapeutic target. PMID- 16357581 TI - Anticholinergic agents in asthma: chronic bronchodilator therapy, relief of acute severe asthma, reduction of chronic viral inflammation and prevention of airway remodeling. AB - PURPOSE OF REVIEW: It is difficult to identify specific groups of asthmatic patients who may benefit from acute or chronic use of anticholinergic agents. Therefore, an important consideration is how anticholinergic agents can be used to achieve clinically effective treatment of asthma. RECENT FINDINGS: A genotype stratified study revealed that greater bronchoprotective effect of anticholinergic agents was observed in asthmatic patients with the Arg/Arg genotype of the beta2-adrenergic receptor. Anticholinergic agents could add to the bronchodilation obtained with beta2-agonists on acute severe asthma. CD8+ T lymphocytes induced by chronic hepatitis C viral infection causes chronic obstructive pulmonary disease-like inflammation in asthma. Virus-specific CD8+ T lymphocytes may induce cholinergic activation in asthma through M2 receptor dysfunction. Therefore, anticholinergic agents are highly effective for asthma associated with chronic viral infection. In contrast, asthma with chronic obstructive pulmonary disease-like inflammation appears to be poorly responsive to beta2-agonists and can lead to partially irreversible airflow limitation. Moreover, a recent report suggested that treatment with inhaled tiotropium bromide markedly inhibited the increase in airway smooth muscle mass, myosin expression, and contractility in asthma. SUMMARY: Anticholinergic agents may benefit stable asthmatics, particularly those who have the Arg/Arg genotype. These agents have a demonstrated role in combination with beta2-agonists in the treatment of acute severe asthma, and may benefit asthmatics with chronic obstructive pulmonary disease-like inflammation. Moreover, these agents could be also beneficial in preventing airway remodeling in asthmatic airways. PMID- 16357582 TI - Urban asthma. AB - PURPOSE OF REVIEW: To review the possible reasons why the prevalence and morbidity of asthma are greater in those living in an urban setting. RECENT FINDINGS: Urban asthma is associated with exposure to air pollution, urban allergens, and violence. Outdoor air pollution is a particular problem in urban areas and affects children more than adults. Asthma has been shown to be exacerbated by ozone, respirable particulates, and nitrogen dioxide. Exposure to high point sources of pollutants such as heavy traffic is of particular concern. Urban allergens include cockroach, mouse, and rat. Cockroach exposure increases the risk of asthma exacerbations and may increase the risk of developing asthma. Although mouse allergen is also found in suburban homes, the concentration is a log-fold higher in inner-city homes at levels known to elicit symptoms in workers in animal facilities. Rat allergen is found in a third of inner-city homes and is associated with asthma morbidity. A recent interventional study showed that comprehensive environmental control of cockroach allergen reduced asthma morbidity. Finally, stress elicits asthma symptoms and exposure to violence is associated with greater asthma morbidity. SUMMARY: The increase in prevalence and morbidity of asthma associated with urban living is associated with at least three plausible causative factors each of which is amenable to intervention, raising the issue of environmental justice issues: controlling air pollution in general and exposure to point sources in particular; reducing cockroach, mice, and rat infestations; and preventing violence. PMID- 16357583 TI - Peak flow monitoring in clinical practice and clinical asthma trials. AB - PURPOSE OF REVIEW: This review summarizes recent reports on peak expiratory flow (PEF) monitoring in clinical asthma trials and clinical practice. RECENT FINDINGS: In clinical trials, summary measures such as average morning PEF provide only a fraction of the available information about asthma control and treatment response. New statistical models should improve the yield from PEF datasets. Improved criteria are needed for the diagnosis of exacerbations, and these may be developed from quality-control analysis of existing datasets. In clinical practice we must reduce the burden of monitoring and increase the ease of interpretation of PEF data. Electronic monitoring, with short message service or Internet communication, may assist with both. There is a need for standardized user-friendly PEF charts and simple statistically appropriate interpretative tools, which will facilitate the development of clinical algorithms and individualized written action plans. Normal values for diurnal variability should be updated to reflect twice daily monitoring. SUMMARY: Current use of PEF data is limited by the burden of monitoring and the continuing use of interpretative tools that were originally developed for their practical feasibility rather than their clinical validity. Both of these problems may be improved by giving attention to methods for recording, displaying and analysing PEF data. PMID- 16357584 TI - Bibliography. Current world literature. Asthma. PMID- 16357585 TI - Looking back, and looking forward. PMID- 16357586 TI - Environmental toxins and neurodegenerative diseases: a challenge for epidemiologists. PMID- 16357587 TI - Socioeconomic context. PMID- 16357588 TI - Diarrhea among young children in an industrialized country. PMID- 16357589 TI - Polychlorinated biphenyls and neurodegenerative disease mortality in an occupational cohort. AB - BACKGROUND: Production of polychlorinated biphenyls (PCBs) ended in the United States in the 1970s, but PCBs persist in the environment and are detectable in the blood of approximately 80% of Americans over age 50. PCBs decrease dopamine levels in rats and monkeys. Loss of dopamine is the hallmark of Parkinson disease, a neurodegenerative disease. There are no epidemiologic studies of PCBs and neurodegenerative disease. METHODS: We conducted a retrospective mortality study of 17,321 PCB-exposed workers to determine whether mortality from Parkinson disease, dementia, and amyotrophic lateral sclerosis was elevated compared with the U.S. population. All workers had a least 90 days employment in 1 of 3 electrical capacitor plants using PCBs from the 1940s to the 1970s. PCB serum levels from a sample of these workers in the 1970s were approximately 10 times the level of community controls. RESULTS: We found no overall excess of Parkinson disease, amyotrophic lateral sclerosis, or dementia in the PCB-exposed cohort (standardized mortality ratios [SMRs]-1.40, 1.11, and 1.26, respectively, and number of deaths-14, 10, and 28 respectively). However, sex-specific analyses revealed that women had an excess of amyotrophic lateral sclerosis (SMR-2.26; 95% confidence interval [CI] = 1.08-4.15; 10 deaths). Furthermore, among highly exposed women (defined by a job-exposure matrix), we found an excess of Parkinson disease (SMR-2.95; 95% CI = 1.08-6.42; 6 deaths) and dementia (SMR-2.04; 95% CI = 1.12-3.43; 14 deaths). CONCLUSIONS: Our data are limited due to small numbers and reliance on mortality rather than incidence data, but are suggestive of an effect of PCBs on neurodegenerative disease for women. The literature does not offer an explanation for why women would be more affected than men by PCB exposure for these outcomes. PMID- 16357590 TI - Neighborhood socioeconomic context, individual income and myocardial infarction. AB - BACKGROUND: The incidence of myocardial infarction (MI) varies among socioeconomic groups, and geographic differences in incidence rates are observed within most urban regions. Whether spatial social differentiation gives rise to social contexts detrimental to health is still an open question. In this study, we evaluate 2 aspects of the neighborhood context as contributory factors in MI: level of economic resources and degree of socioeconomic homogeneity. We adopt a multilevel approach to analyze potential mechanisms, which involve individual social characteristics. METHODS: We analyzed data from the SHEEP study, a population-based case-control study of first events of acute MI in Stockholm County in 1992-1994. Data on socioeconomic characteristics in neighborhoods came from total population registers of income and social circumstances. RESULTS: The level of neighborhood socioeconomic resources had a contextual effect on the relative risk of MI after adjustment for individual social characteristics. The incidence rate ratio (IRR) in low-income, compared with high-income, neighborhoods was 1.88 for women and 1.52 for men. Although the degree of socioeconomic homogeneity in neighborhoods has less impact on MI, the IRR for men in homogenous low-income areas compared with men living in heterogeneous high income areas was 2.65. For men, the combined exposure to low-personal disposable income and low-income level in the neighborhood seemed to have an additive effect but for women, a synergistic (supra-additive) effect was found. CONCLUSION: The socioeconomic context of neighborhoods has an effect on cardiovascular outcomes. PMID- 16357591 TI - Risk factors for diarrhea among children in an industrialized country. AB - BACKGROUND: Risk factors for childhood diarrhea in industrialized countries are not well characterized, although diarrhea remains an important cause of morbidity. METHODS: We conducted a case-control study of 422 cases and 866 controls over 22 months in Denmark. We selected cases among children under 5 years of age with diarrhea. Age-matched healthy controls were selected from the background population using a population register. Parents were interviewed about possible exposures and underlying conditions. In addition, stool samples from both cases and controls were analyzed for viruses, parasites, and bacteria. We analyzed risk factors for diarrhea in general and for diarrhea of a viral, bacterial, or "unknown" etiology using logistic regression. RESULTS: The following factors were independently associated with an increased risk of diarrhea: recent foreign travel, contact with symptomatic persons (particularly in daycare centers), hospitalization, contact with a dog with diarrhea, private daycare, consumption of products containing formula milk, unemployment and low educational status of parents, and prior diagnosis of several types of atopic diseases. In a pathogenic-specific analysis of diarrhea of bacterial (73 patients), viral (88), or "unknown" (222) etiology, the major risk factor for viral diarrhea was contact with symptomatic persons. For bacterial diarrhea, foreign travel and socioeconomic factors were the main risk factors. CONCLUSIONS: Viral diarrhea appears to be transmitted predominantly from person to person, whereas bacterial diarrhea appears to be primarily foodborne. A substantial portion of the diarrheal episodes may be of noninfectious etiology. Limiting child-to-child transmission of disease in daycare centers may substantially reduce the disease burden. PMID- 16357592 TI - Perceived exertion in physical activity and risk of gestational diabetes mellitus. AB - BACKGROUND: Physical activity has been associated with decreased risk of gestational diabetes mellitus. Expanding on 2 previously published analyses of absolute exertion measures (time spent and energy expended), we assessed the relation between perceived exertion during usual prepregnancy recreational physical activity and gestational diabetes. METHODS: We analyzed data from a Washington State 1998-2002 case-control study (216 cases, 472 controls) and a 1996-2002 prospective cohort study (897 participants) separately. We used logistic regression models to derive odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Among case-control participants, risk of gestational diabetes was markedly lower for women who reported very strenuous to maximal exertion in usual activity during the year before pregnancy versus those who reported negligible or minimal exertion (adjusted OR = 0.19; CI = 0.15-0.50). There was a direct inverse relation between perceived exertion and risk of gestational diabetes. This relation was also evident among the subset of participants who did not meet physical activity guidelines in the year before pregnancy. Similarly, the OR among cohort participants reporting very strenuous to maximal exertion was 0.57 (0.24-1.37) versus those reporting negligible to moderate exertion. CONCLUSIONS: These results suggest that risk of gestational diabetes is inversely related to the exertion perceived during recreational physical activity in the year before pregnancy. Perceived exertion may be a valuable addition to behavior and fitness measures in assessing relations between physical activity and pregnancy-related health outcomes. PMID- 16357594 TI - Smokeless tobacco use and risk of stillbirth: a cohort study in Mumbai, India. AB - BACKGROUND: Maternal cigarette smoking has been causally associated with an increased risk for stillbirth. Preliminary reports suggest an increased risk for stillbirth with smokeless tobacco use during pregnancy. METHODS: We conducted a population-based prospective cohort study to investigate this association by using a house-to-house approach to recruit 1,217 women who were between 3 and 7 months' gestation. Of these, 96% were contacted after delivery to determine the pregnancy outcome. Demographic and maternal variables which were apparently associated either with stillbirth or with smokeless tobacco use (OR >or= 1.5) were included as potential confounders. Stillbirth was defined as any delivery of a dead fetus after 20 completed weeks of gestation. We used time-to-event methods to analyze the risk of stillbirth. RESULTS: Overall occurrence of stillbirth among singleton deliveries in this population was 4.1%. Smokeless tobacco use was reported by 17% of women; 8.9% of smokeless tobacco users had a stillbirth compared with 3.1% among nonusers (life-table adjusted hazard ratio = 3.1; 95% confidence interval = 1.7-5.6). After adjustment by the Cox proportional hazards procedure for age, educational and socioeconomic background, working status of mother, parity, prenatal care variables, and place of delivery, the risk for stillbirth in users was 2.6 (95% confidence interval-1.4-4.8). Most women used mishri (a pyrolyzed tobacco product often used as dentifrice), and there was a dose-response relationship between the daily frequency of use and stillbirth risk. The risk of stillbirth associated with smokeless tobacco use was greater in earlier gestational periods. CONCLUSIONS: Smokeless tobacco use during pregnancy increases stillbirth risk, with a risk at least as great as that associated with maternal cigarette smoking. PMID- 16357593 TI - Xenobiotic-metabolizing genes and small-for-gestational-age births: interaction with maternal smoking. AB - BACKGROUND: Little is known about the role of xenobiotic-metabolizing gene variants as risk factors for small-for-gestational-age (SGA) births or as modifiers for the effects of exposures such as maternal smoking. METHODS: We conducted 2 joint studies: a case-control design including 493 cases (birth weight below the 10th percentile according to gestational age and sex) and 472 controls (at or above the 10th percentile) and a family-based study (mother, father, and newborn) with approximately 250 case trios and a similar number of control trios. Logistic regression and a log-linear model were used to analyze the association between genetic variants such as CYP1A1*2A, CYP1A1*2B, CYP1A1*4, GSTT1, GSTM1, and XRCC3 and SGA. The interaction between genetic variants and maternal smoking was also studied. RESULTS: The odds ratio (OR) for the association of complete maternal GSTT1 deletion with SGA was 0.63 (95% confidence interval = 0.41-0.97), and that for the complete newborn GSTM1 deletion was 0.74 (0.55-0.98). Newborns with the partial GSTT1 deletion had an OR of 1.40 (1.01 1.95), and newborns homozygous for CYP1A1*2A had an OR of 4.28 (1.02-18.0). These results were coherent with the trio-based results. Significant interactions were observed between maternal smoking in the third trimester and CYP1A1*2A (P = 0.03), XRCC3 (P = 0.03), and newborn GSTT1 (P = 0.01). CONCLUSIONS: Certain genetic variants involved in the metabolism of xenobiotics increase the risk of SGA, as well as modify the effects of maternal smoking by increasing or decreasing its risk. PMID- 16357595 TI - Menstrual cycle characteristics: associations with fertility and spontaneous abortion. AB - BACKGROUND: Epidemiologists often use menstrual cycle patterns as indicators of endocrine function in environmental and occupational studies, yet few studies have considered whether menstrual cycle characteristics are associated with fertility or pregnancy outcome. METHODS: We prospectively studied 470 women to determine whether cycle length or bleed length were associated with fertility or spontaneous abortion. Women completed daily diaries with information on menstrual bleeding, intercourse, birth control use, and covariates. For each menstrual cycle, women collected at least 2 urine samples, which were assayed for human chorionic gonadotropin to define early pregnancies. Women were followed for 1 year or until the end of a clinical pregnancy. RESULTS: Cycles with lengths of 30 to 31 days preceded cycles with the highest fecundity. Shorter cycles were less likely to be followed by conception (fecundity ratio [FR] = 0.6; 95% confidence interval [CI] = 0.4-1.0). Compared with 30- to 31-day cycles, conceptions after shorter and longer cycles were more likely to be spontaneously aborted (for shorter cycles, odds ratio [OR] = 3.0 [95% CI = 0.9-9.6] and for longer cycles, OR = 3.0 [0.9-10.6]). Cycles with 5 days of menstrual bleeding had the highest fecundity. Cycles with up to 4 days of bleeding had lower fecundity (for bleed lengths of 4 days, FR = 0.5 [0.3-0.8] and for bleed lengths less than 4 days, FR = 0.6 [0.3-0.9]). Spontaneous abortion was less likely after bleeds greater than 5 days (OR = 0.4 [0.1-1.1]) when compared with 5-day bleeds. CONCLUSIONS: Menstrual cycle characteristics appear to be associated with fertility and spontaneous abortion. PMID- 16357596 TI - Exposure to nonpersistent insecticides and male reproductive hormones. AB - BACKGROUND: Urinary metabolites of several nonpersistent insecticides have been measured in a high percentage of men in the general population, suggesting widespread environmental exposures to these compounds. The present study explored the association of urinary concentrations of 3,5,6-trichloro-2-pyridinol (TCPY), a metabolite of chlorpyrifos and chlorpyrifos-methyl, and 1-naphthol (1N), a metabolite of carbaryl and naphthalene, with serum reproductive hormone levels in adult men. METHODS: Subjects (n = 268) were the male partners in couples presenting to a Massachusetts infertility clinic in years 2000 through 2003. TCPY and 1N were measured in a spot urine sample from each subject and adjusted for dilution using specific gravity. Reproductive hormones (follicle-stimulating hormone, leuteinizing hormone, inhibin B, testosterone, and sex hormone-binding globulin) were measured in serum collected from subjects during the same clinic visit. RESULTS: Multiple linear regression models showed an inverse association between TCPY and testosterone concentration. An interquartile range (IQR) increase in TCPY was associated with a decline of 25 ng/dL (95% confidence interval = -40 to -10) in testosterone concentration. The association appeared to be dose-dependent when exposure was divided into quintiles. The highest TCPY quintile was associated with a testosterone decline of 83 ng/dL (-128 to -39) compared with the lowest TCPY quintile. We also found inverse associations between TCPY and free androgen index and between 1N and testosterone, and suggestive inverse associations between TCPY and leuteinizing hormone and between 1N and free androgen index. CONCLUSION: In adult men, TCPY and 1N were associated with reduced testosterone levels. On a population level, these reductions are of potential public health importance because of widespread exposure to these nonpersistent insecticides. PMID- 16357597 TI - Exposure misclassification in studies of agricultural pesticides: insights from biomonitoring. AB - BACKGROUND: Epidemiologists often assess lifetime pesticide exposure by questioning participants about use of specific pesticides and associated work practices. Recently, Dosemeci and colleagues proposed an algorithm to estimate lifetime average exposure intensity from questionnaire information. We evaluated this algorithm against measured urinary pesticide concentrations for farmers who applied glyphosate (n = 48), 2,4-D (n = 34), or chlorpyrifos (n = 34). METHODS: Algorithm scores were calculated separately based on trained field observers' and farmers' evaluations of application conditions. Statistical analyses included nonparametric correlations, assessment of categorical agreement, and categorical evaluation of exposure distributions. RESULTS: Based on field observers' assessments, there were moderate correlations between algorithm scores and urine concentrations for glyphosate (r = 0.47; 95% confidence interval [CI] = 0.21 to 0.66) and 2,4-D (0.45; 0.13 to 0.68). Correlations were lower when algorithm scores were based on participants' self-reports (for glyphosate, r = 0.23 [CI = 0.07 to 0.48]; for 2,4-D, r = 0.25 [-0.10 to 0.54]). For chlorpyrifos, there were contrasting correlations for liquid (0.42; 0.01 to 0.70) and granular formulations (-0.44; -0.83 to 0.29) based on both observers' and participants' inputs. Percent agreement in categorical analyses for the 3 pesticides ranged from 20% to 44%, and there was appreciable overlap in the exposure distributions across categories. CONCLUSIONS: Our results demonstrate the importance of collecting type of pesticide formulation and suggest a generic exposure assessment is likely to result in appreciable exposure misclassification for many pesticides. PMID- 16357598 TI - Impact of the 2003 heatwave on all-cause mortality in 9 French cities. AB - BACKGROUND: A heatwave occurred in France in August 2003, with an accompanying excess of all-cause mortality. This study quantifies this excess mortality and investigates a possible harvesting effect in the few weeks after the heatwave. METHODS: A time-series study using a Poisson regression model with regression splines to control for nonlinear confounders was used to analyze the correlation between heatwave variable and mortality in 9 French cities. RESULTS: After controlling for long-term and seasonal time trends and the usual effects of temperature and air pollution, we estimated that 3,096 extra deaths resulted from the heatwave. The maximum daily relative risk of mortality during the heatwave (compared with expected deaths at that time of year) ranged from 1.16 in Le Havre to 5.00 in Paris. There was little evidence of mortality displacement in the few weeks after the heatwave, with an estimated deficit of 253 deaths at the end of the period. CONCLUSIONS: The heatwave in France during August 2003 was associated with a large increase in the number of deaths. The impact estimated using a time series design was consistent with crude previous estimates of the impact of the heatwave. This finding suggests that neither air pollution nor long-term and seasonal trends confounded previous estimates. There was no evidence to suggest that the extras deaths associated with the heatwave were simply brought forward in time. PMID- 16357599 TI - Estrogen metabolism and breast cancer. AB - BACKGROUND: Specific pathways involved in estrogen metabolism may play a role in the etiology of breast cancer. We used data from a large population-based case control study to assess the association of the urinary estrogen metabolites 2 hydroxyestrone (2-OHE1), 16alpha-hydroxyestrone (16-OHE1), and their ratio (2/16) with both invasive and in situ breast cancer. METHODS: Study participants from the Long Island Breast Cancer Study Project provided a spot urine specimen and completed a comprehensive interviewer-administered questionnaire. Women who used exogenous hormones or who took tamoxifen in the 6 months before urine collection were excluded from the analysis, leaving 269 invasive cases, 158 in situ cases, and 326 controls. Unconditional logistic regression was used to obtain adjusted odds ratios (ORs) for invasive and in situ breast cancer, separately, in relation to tertiles of the individual metabolites (standardized for creatinine) and the 2/16 ratio, stratified by menopausal status. RESULTS: The OR for invasive breast cancer was inversely associated with the 2/16 ratio among premenopausal women (OR = 0.50 for extreme tertiles; 95% confidence interval = 0.25-1.01). ORs ranged from 0.32 to 0.60 when women were stratified by whether cases had received chemotherapy within 6 months before urine collection and by estrogen receptor status. In postmenopausal women, there was a slight reduction in the odds ratio for invasive cancer with high levels of the 2/16 ratio (OR = 0.78; 95% confidence interval = 0.46-1.33). Neither the individual metabolites nor the ratio were associated with in situ breast cancer. CONCLUSION: These data provide support for the hypothesis that the 2/16 ratio is associated with reduced breast cancer risk. The most consistent associations were observed with invasive cancer in premenopausal women. PMID- 16357600 TI - EPHX1 polymorphisms and the risk of lung cancer: a HuGE review. AB - BACKGROUND: Microsomal epoxide hydrolase 1 (EPHX1) plays an important role in both the activation and detoxification of tobacco-derived carcinogens. Polymorphisms at exons 3 and 4 of the EPHX1 gene have been reported to be associated with variations in EPHX1 activity. The aim of this study is to review and summarize the available molecular epidemiologic studies of lung cancer and EPHX1. METHODS: We searched MEDLINE, Current Contents, and Web of Science databases for studies published before August 2004. We conducted a systematic review and meta-analysis of 13 case-control studies. Summary odds ratios and summary prevalence of the variant allele (genotype) of both polymorphisms in the EPHX1 gene were calculated using the DerSimonian and Laird method. RESULTS: The low-activity (variant) genotype of EPHX1 polymorphism at exon 3 was associated with decreased risk of lung cancer (odds ratio = 0.65; 95% confidence interval = 0.44-0.96) in lung cancer risk among whites. In white populations, the high activity (variant) genotype of EPHX1 polymorphism at exon 4 was associated with a modest increase in risk of lung cancer (1.22; 0.79-1.90) and the predicted low activity was associated with a modest decrease in risk (0.72; 0.43-1.22). CONCLUSIONS: EPHX1 enzyme may act as a phase I enzyme in lung carcinogenesis. The low-activity genotype of EPHX1 gene is associated with decreased risk of lung cancer among whites. PMID- 16357601 TI - A comparison of underlying cause and multiple causes of death: US vital statistics, 2000-2001. AB - BACKGROUND: Mortality statistics can be compiled using underlying cause-of-death data or multiple cause-of-death data, which include other contributing causes of death. METHODS: For the leading causes of death in the United States during 2000 2001, we compared underlying and multiple cause-of-death statistics. RESULTS: For some conditions, little difference was observed between the 2 estimates. For other conditions, up to 10 times more deaths were identified from multiple-cause data than from underlying-cause data. The 10 leading causes of death differed when using the 2 types of data. CONCLUSIONS: Whenever possible, underlying and multiple cause-of-death statistics should both be presented. Analyses that use only the underlying cause of death ignore additional information that is readily available from multiple-cause data, and the more limited data may underestimate the importance of several leading causes of death. PMID- 16357602 TI - Analgesic drug use and risk of ovarian cancer. AB - BACKGROUND: Previous epidemiologic research suggests that analgesic use may reduce the risk of ovarian cancer, although results are not consistent. METHODS: In a population-based, case-control study, we analyzed data from 586 ovarian cancer cases and 627 matched controls in North Carolina for the relationship between analgesic use and ovarian cancer risk. Logistic regression analysis was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) while adjusting for potential confounders. RESULTS: Use of any nonsteroidal antiinflammatory drugs, including aspirin, within 5 years of diagnosis/interview was found to be associated with a reduction in the risk of ovarian cancer (adjusted OR = 0.72; 95% CI = 0.56-0.92). For use of acetaminophen, the OR was 0.78 (95% CI = 0.56-1.08). CONCLUSIONS: These data support an inverse relationship between the use of both nonsteroidal antiinflammatory drugs and acetaminophen and the risk of ovarian cancer. PMID- 16357603 TI - Night work and risk of breast cancer. AB - BACKGROUND: Melatonin shows potential oncostatic activity and is acutely suppressed by light exposure. Some evidence suggests an association between night work and breast cancer risk, possibly through the melatonin pathway. METHODS: In a cohort of premenopausal nurses, we prospectively studied the relation between rotating night shift work and breast cancer risk. Total number of months during which the nurses worked rotating night shifts was first assessed at baseline in 1989 and periodically updated thereafter. We used Cox proportional hazards models to calculate relative risks (RRs) and 95% confidence intervals (CIs). RESULTS: Among 115,022 women without cancer at baseline, 1,352 developed invasive breast cancer during 12 years of follow up. Women who reported more than 20 years of rotating night shift work experienced an elevated relative risk of breast cancer compared with women who did not report any rotating night shift work (multivariate RR = 1.79; 95% CI = 1.06-3.01). There was no increase in risk associated with fewer years of rotating night work. CONCLUSION: Our results suggest a modestly elevated risk of breast cancer after longer periods of rotating night work. Additional studies are warranted to rule out small sample size or uncontrolled sources for confounding as alternative explanations. PMID- 16357604 TI - Breast-feeding and overweight in adolescence: within-family analysis [corrected]. AB - BACKGROUND: Previous reports have found associations between having been breast fed and a reduced risk of being overweight. These associations may be confounded by sociocultural determinants of both breast-feeding and obesity. We addressed this possibility by assessing the association of breast-feeding duration with adolescent obesity within sibling sets. METHODS: We surveyed 5,614 siblings age 9 to 14 years and their mothers. These children were a subset of participants in the Growing Up Today Study, in which we had previously reported an inverse association of breast-feeding duration with overweight. We compared the prevalence of overweight (body mass index exceeding the age-sex-specific 85th percentile) in siblings who were breast-fed longer than the mean duration of their sibship with those who were breast-fed for a shorter period. Then we compared odds ratios from this within-family analysis with odds ratios from an overall (ie, not within-family) analysis. RESULTS: Mean +/- standard deviation breast-feeding duration was 6.4 +/- 4.0 months, and crude prevalence of overweight was 19%. On average, siblings who were breast-fed longer than their family mean had breast-feeding duration 3.7 months longer than their shorter duration siblings. The adjusted odds ratio (OR) for overweight among siblings with longer breast-feeding duration, compared with shorter duration, was 0.92 (95% confidence interval = 0.76-1.11). In overall analyses, the adjusted OR was 0.94 (0.88-1.00) for each 3.7-month increment in breast-feeding duration. CONCLUSION: The estimated OR for the within-family analysis was close to the overall estimate, suggesting that the apparent protective effect of breast feeding on later obesity was not highly confounded by unmeasured sociocultural factors. A larger study of siblings, however, would be needed to confirm this conclusion. PMID- 16357605 TI - "East side story": on being an epidemiologist in the former USSR: an interview with Marcus Klingberg. AB - Marcus Klingberg was born on 7 October 1918, in Warsaw, Poland, into a Hasidic, rabbinical family. After the Nazi invasion of Poland, he escaped to the USSR where he trained and worked as an epidemiologist from 1939 to 1945. For 35 years after the war, he continued his professional work in Israel. The harsh conditions within the Soviet Union during World War II provided a challenging setting for epidemiologic work-a setting that has remained largely hidden from Western view. In this interview, Klingberg describes his work as an epidemiologist in the USSR and his subsequent encounter with Western epidemiology. PMID- 16357606 TI - Incidence of autism over time. PMID- 16357608 TI - Indoor radon and lung cancer. PMID- 16357611 TI - Body mass regulation at altitude. AB - Altitude exposure decreases energy intake and thus induces an energy deficit resulting in the loss of body mass. The energy deficit is worsened when energy expenditure is increased by exercise because the increase is not matched by an increased energy intake. The indicated fuel for the optimal use of the rarefied oxygen at altitude is carbohydrate. For optimal taste and maximizing energy intake, fat is the best. Protein should rather be limited because of its high thermic effect. Intestinal malabsorption probably does not play a role in the energy deficit. Even though the sensation of exertion during climbing at high altitude is intense, the actual energy expenditure is not high. PMID- 16357612 TI - Weight loss at high altitude: pathophysiology and practical implications. AB - Climbers at high altitude (>5000 m) lose weight. This impairs performance and safety, but the mechanisms are not entirely due to an imbalance between energy intake and expenditure. There is some evidence of carbohydrate malabsorption, but there are also changes in fat metabolism and total body water. This paper considers the physiological control of weight and then discusses the changes in each parameter before addressing the practical implications. PMID- 16357613 TI - Combining infliximab with methotrexate for the induction and maintenance of remission in refractory Crohn's disease: a controlled pilot study. AB - OBJECTIVES: Immunosuppression of chronic active Crohn's disease resistant or intolerant to purine antimetabolites still remains a clinical challenge. To obtain long-lasting effects with the anti-TNF-alpha antibody infliximab repeated infusions are often required. Methotrexate has been shown to be a moderately effective drug in maintaining remission in Crohn's disease. The aim of the present pilot study was to evaluate the combination of infliximab and methotrexate as therapy for refractory Crohn's disease. METHODS: Nineteen patients with chronic active Crohn's disease resistant or intolerant to azathioprine were enrolled. Patients received either two infusions of infliximab (5 mg/kg) alone (n=8) or in combination with long-term methotrexate at a dosage of 20 mg/week (n=11) over 48 weeks. RESULTS: Two out of eight patients receiving infliximab monotherapy and four out of 11 patients treated with infliximab and concomitant methotrexate had discontinued study treatment by week 48, solely because of lack of efficacy. Clinical remission at week 48 was observed in five out of seven patients treated with infliximab and methotrexate, but only in two out of six patients receiving infliximab monotherapy. In addition, patients treated with concomitant methotrexate achieved remission earlier (median time 2 versus 18 weeks) and needed fewer steroids (median prednisolone dose 0 versus 11.8 mg). Despite an increased mean number of adverse events per patient in the methotrexate group, the proportions of patients experiencing any adverse events and serious adverse events were similar across treatment groups. CONCLUSIONS: The combination of infliximab with long-term methotrexate may be a promising concept in refractory Crohn's disease. Our data prompt larger trials. PMID- 16357614 TI - Re-introduction of azathioprine in previously intolerant patients. AB - BACKGROUND AND AIMS: Azathioprine is an important steroid sparing agent in the management of patients with inflammatory bowel disease. However, side effects are a problem in a significant minority of patients. We proposed that desensitization might increase the number of patients able to tolerate the drug. METHODS: Successive inflammatory bowel disease patients who were intolerant to azathioprine in our hospital gastroenterology clinic were invited to recommence the drug at a low dose, gradually building up to a therapeutic dose. Patients were observed for the recurrence of side effects. Patients who did not wish to participate were offered alternative immunosuppressant therapy. RESULTS: Fourteen patients elected to attempt desensitization. Nine of these (64%) were able to tolerate a full dose of azathioprine. The remainder suffered a return of their side effects and were offered alternative treatment. CONCLUSIONS: Azathioprine can be tolerated in some people who have been previously intolerant by the use of desensitization. PMID- 16357615 TI - Apoptosis and Bcl-2 expression in the livers of patients with steatohepatitis. AB - OBJECTIVES: Apoptosis may play a role in the pathogenesis of alcoholic (ASH) and non-alcoholic steatohepatitis (NASH). In this study, we investigated the modulation of apoptosis-related liver proteins in steatohepatitis. METHODS: Hepatocyte apoptosis was evaluated by the TUNEL assay in liver tissue of 12 patients with NASH, 12 with ASH and in histologically normal controls. In addition, caspase-3 processing was evaluated by immunoblot analysis. Expression of death receptors, Bcl-2 family members, and NF-kappaB inhibitor (IkappaB) were determined by western blot. Liver biopsies were also graded for inflammation and fibrosis. RESULTS: Apoptotic hepatocytes were markedly increased in NASH (P<0.05) and ASH (P<0.001) as compared to controls. Active caspase-3 was also elevated in steatohepatitis (P<0.01), coinciding with upregulation of pro-apoptotic Bax (P<0.001). Further, production of tumour necrosis factor-receptor 1 was increased up to 4-fold (P<0.05). Degradation of IkappaB increased >70% in steatohepatitis (P<0.001). Notably, Bcl-2 was also strongly expressed (>100-fold; P<0.001). These data were significantly correlated with relative degrees of portal and lobular inflammation. CONCLUSION: The results show that liver injury in NASH and ASH is associated with apoptosis and NF-kappaB activation. Anti-apoptotic Bcl-2 is strongly expressed, probably reflecting an adaptive response to obesity or alcohol-related stress. PMID- 16357616 TI - Sleep apnoea syndrome and early stage cirrhosis: a pilot study. AB - OBJECTIVES: Hepatic encephalopathy in patients with end-stage liver cirrhosis is associated with alterations in sleep patterns. Cirrhosis may also affect pulmonary function and it might be involved in the development of obstructive sleep apnoea syndrome (OSAS) in patients with ascites. We carried out a study to evaluate the presence of OSAS in cirrhotic patients without evidence of ascites (early stage cirrhosis). METHODS: We investigated 20 patients with Child A or B cirrhosis (19 and one, respectively) and 10 non-cirrhotic patients with chronic viral hepatitis (disease control group). All subjects were interviewed and underwent a thorough physical examination, a full polysomnographic study and a pulmonary function testing by spirometry. Serum samples were also obtained in order to determine the liver function tests. RESULTS: The presence of OSAS and inverted sleep patterns was similar in cirrhotic patients and disease controls. However, significant correlations were revealed between age and hypopnoeas per hour of sleep; age and the Apneas/Hypopneas Index (AHI); age and FEV1/FVC; gamma glutamyl transpeptidase and FEV1/FVC; and total bilirubin and total sleep time. CONCLUSIONS: Early stage cirrhosis is not associated with sleep disorders and OSAS. However, total bilirubin levels might be a useful laboratory marker for early assessment of disturbance in sleep patterns and therefore of subclinical hepatic encephalopathy in Child A cirrhosis. PMID- 16357617 TI - Correlation of ultrasound attenuation and histopathological parameters of the liver in chronic diffuse liver diseases. AB - OBJECTIVE: In patients with chronic diffuse liver diseases two characteristic ultrasound patterns of bright liver, the low (DI) and the high (DII) attenuation types, are seen. Correlation was studied between liver attenuation and histopathological analysis of biopsy specimens of the same patients. METHODS: Ultrasound attenuation of the liver was measured quantitatively using a homogeneous tissue-equivalent reference phantom. Semiquantitative scores were used for histopathological analysis of biopsy specimens. RESULTS: One hundred and twenty-two patients were investigated; 40 of them showed a normal liver echopattern. The average attenuation was 0.68+/-0.03 dB/cm per MHz. Histopathological parameters were normal in most of the patients, except for four cases where a minimal amount of collagen and 14 cases where a subtle amount of lipid content could be detected, in three cases accompanied by some cells indicating inflammation. From 82 patients with bright liver, 47 showed the DI type. The average attenuation was 0.80+/-0.03 dB/cm per MHz. In all of these patients, significant increases of collagen content and inflammatory reaction were found. In 23 cases a negligible amount of lipid deposition could also be revealed. Thirty-five patients exhibited a DII-type bright liver. The average attenuation was 1.21+/-0.06 dB/cm per MHz. A significant amount of lipid deposition was detected in all cases. In 13 patients a minimal amount of collagen and in 14 patients some inflammatory cells were detected. CONCLUSIONS: In livers with a normal echopattern, none or minimal pathological changes were seen. In DI type bright liver, connective tissue dominance exists, in DII-type bright liver lipid deposition dominance. On this basis it is proved that the diagnosis of a fatty liver can be established without biopsy, if no other indication for biopsy exists. PMID- 16357618 TI - Late onset of Wilson's disease in a family with genetic haemochromatosis. AB - We report the coexistence of Wilson's disease and genetic haemochromatosis in one family. The diagnosis of genetic haemochromatosis was established in a 52-year old man. Among his siblings, one 57-year-old sister and one 55-year-old brother had decreased copper and ceruloplasmin levels in serum and increased urinary copper excretion. The sister shared the same human leucocyte antigen haplotypes and was homozygous for the HFE mutation C282Y, like the propositus. However, she had normal liver iron content and increased liver copper content. Her dietary copper intake was probably excessive. The association of Wilson's disease and genetic haemochromatosis is rare and has only been described twice. The onset of Wilson's disease after 50 years of age is rare; Wilson's disease should be considered in any patient with unexplained chronic liver disease; an excess in liver copper content might be induced by excessive dietary input in a susceptible individual. PMID- 16357619 TI - Pharmacologic treatment of portal hypertension in the prevention of community acquired spontaneous bacterial peritonitis. AB - INTRODUCTION: Given that beta-blockers reduce the incidence of bacterial translocation in cirrhotic rats, the aim of this study was to compare the long term incidence of spontaneous bacterial peritonitis in cirrhotic patients submitted to pharmacologic versus endoscopic treatment to prevent variceal rebleeding. PATIENTS AND METHODS: Two hundred and thirty patients with variceal hemorrhage were included in two previous randomized trials performed to compare the efficacy of medication (nadolol plus isosorbide mononitrate, n=115) versus endoscopic treatment (n=115) with sclerotherapy or ligation for the prevention of rebleeding. RESULTS: The mean follow-up was 23+/-1.4 months. The characteristics of the patients and the number of patients on long-term prophylaxis with norfloxacin were similar in both groups. The incidence of spontaneous bacterial peritonitis was lower in the medication group (9 versus 14.7%, P=NS). The probability of spontaneous bacterial peritonitis was also lower in the medication group (6 versus 12% at 1 year, 22 versus 36% at 5 years; P=0.08), due to a significantly lower probability of community-acquired spontaneous bacterial peritonitis in this group (1 versus 10% at 1 year, 18 versus 32% at 5 years; P=0.02). Patients with no hemodynamic response to therapy had a significantly higher probability to develop community-acquired spontaneous bacterial peritonitis during follow-up than hemodynamic responders (P<0.03). Long-term probability of developing community-acquired spontaneous bacterial peritonitis is lower in patients submitted to pharmacologic treatment for preventing variceal rebleeding than in those submitted to endoscopic treatment. CONCLUSION: Long-term pharmacologic prophylaxis of variceal rebleeding contributes to the prevention of community-acquired spontaneous bacterial peritonitis. PMID- 16357620 TI - A predictive index for the diagnosis of cirrhosis in hepatitis C based on clinical, laboratory, and ultrasound findings. AB - OBJECTIVE: To develop and validate a non-invasive index to predict the presence of cirrhosis in patients with chronic hepatitis C on the basis of clinical, laboratory, and ultrasound findings. MATERIALS AND METHODS: Data from the complete history and physical examination, serologic studies, liver ultrasound, and liver biopsy of patients with chronic hepatitis C were analyzed using multivariate regression to develop a cirrhosis predictive index. This index was then applied prospectively to another group of patients with chronic hepatitis C to determine its accuracy. RESULTS: Three hundred and thirty-two patients were included (mean age, 48.5+/-18.7 years; male-female ratio, 1.27). Sixty-seven patients (20%) had cirrhosis at histology. Logistic regression identified seven variables that predicted cirrhosis: age>or=60 years, platelet countor=1, prothrombin time (Ratio)>or=1.1, caudate hypertrophy, right lobe atrophy and splenomegaly. Patients scoring>or=22 in total had a statistically significant probability of cirrhosis (sensitivity, 80%; specificity, 96%; and diagnostic accuracy, 94%). CONCLUSION: Cirrhosis can be predicted in patients with chronic hepatitis C by the evaluation of seven clinical, laboratory, and sonographic variables. The index will be useful for the management and follow-up of hepatitis C patients drastically reducing the indications for biopsy in this context. PMID- 16357621 TI - Tolerance to gastric distension in patients with functional dyspepsia: modulation by a cholinergic and nitrergic method. AB - BACKGROUND: In patients with functional dyspepsia (FD), tolerance to gastric fundus distension is frequently compromised. We tested the contribution of contractile cholinergic or relaxing nitrergic pathways in this disorder. METHODS: Thirty-nine patients (29 women, 10 men) with FD participated in this study. Gastric distension studies where carried out by inflating a fundic balloon until the maximum distension volume (MDV) tolerated by the patient. Gastric distension was first evaluated in basal control conditions before repeating the test after the injection of anticholinergic hyoscine (Buscopan 20 mg intravenous) or following the administration of nitric oxide donor nitroglycerin (Nitrolingual 1.2 mg sublingual). RESULTS: MDVs were significantly (P<0.01) lower in FD patients (495+/-27 ml) than in controls (995+/-59 ml). Intolerance to fundic distension was found in 77% of dyspepsia patients. Hyoscine improved the threshold for discomfort (343+/-21 versus 421+/-43 ml; P<0.05), as well as the MDV (510+/-36 versus 635+/-44 ml; P<0.01). This drug effect was seen in 71% (10/14) of patients. Nitroglycerin failed to modify thresholds. However, improved tolerance was seen in 15% (2/13) of the patients. CONCLUSION: Intolerance to fundic distension is a frequent finding in FD (77% patients). The cholinergic pathway seems to be a predominant factor involved in tolerance to distension in patients with FD. Fundus-relaxing drugs should be considered for the treatment of dyspepsia. PMID- 16357622 TI - The role of dominant stenoses in bacterial infections of bile ducts in primary sclerosing cholangitis. AB - OBJECTIVE: Primary sclerosing cholangitis (PSC) is characterized by progressive fibrotic inflammation and strictures of the biliary system. We studied the role of dominant stenoses in bacterial biliary infections and the effect of routine antibiotic administration with cholangiography. DESIGN: A prospective clinical trial without blinding or randomization. SETTING: The endoscopy unit in a university hospital. PARTICIPANTS: Fifty patients with PSC entered and finished the study. INTERVENTIONS: A total of 103 endoscopic retrograde cholangiographies (ERC) was performed in 37 PSC patients with dominant stenosis and 13 controls with PSC but no dominant stenosis. After selective cannulation of the bile duct, bile samples were obtained during each procedure. All patients received systemic antibiotic treatment with ciprofloxacin for one week after ERC. RESULTS: Enteric bacteria were detected in the bile specimens of 15 out of 37 PSC patients (40.5%) with dominant stenosis but never in the absence of dominant stenosis (P=0.004). Positive cultures for enteric bacteria were associated with elevated serum C reactive protein, high leukocyte counts in bile (P<0.05) and the deterioration of liver function assessed by increasing bilirubin levels during the follow-up period lasting a median of 7 months (P=0.06). Despite the high rate of susceptibility in vitro, ciprofloxacin treatment eradicated enteric bacteria in only two out of 12 cases. CONCLUSION: Bacterial infection of the bile ducts with dominant stenosis is a frequent finding and may play a role in the progression of PSC. Short-course antibiotic treatment is not very effective in eradicating acteria from the bile ducts. PMID- 16357623 TI - Disappearance of anti-Saccharomyces cerevisiae antibodies in coeliac disease during a gluten-free diet. AB - BACKGROUND: Anti-Saccharomyces cerevisiae antibodies (ASCAs) are known to be positive in about 65% of Crohn's disease patients, in up to 43% of coeliac disease patients and in 0-5% of healthy controls. Coeliac disease might be an in vivo model for unravelling the role of mucosal integrity in the formation of ASCAs since mucosal abnormalities normalize during a gluten-free diet (GFD). AIMS: Firstly, to evaluate, retrospectively, the frequency of ASCA positivity in coeliac patients both at diagnosis and during follow-up on a GFD. Secondly, to study the correlation between ASCA positivity and mucosal damage. METHODS: One hundred and eleven patients with histologically proven coeliac disease, positive endomysium antibodies on diagnosis and normalization of trans-glutaminase antibodies (t-TGAs) after successful adherence to a GFD were included. ASCAs, IgA and IgG were tested by enzyme-linked immunosorbent assays both at diagnosis and after the GFD. RESULTS: Eighty-three children and 28 adults were included in this study. The mean age at diagnosis was 4.6 years for children and 48 years for adults. At diagnosis 15/83 (18%) of children were ASCA positive (either IgG or IgA), compared to 17/28 (61%) of adults. After successful adherence to a GFD and normalization of t-TGAs only one child remained ASCA positive (1%) compared to eight adults (29%). Two out of 28 (7%) adults remained positive for both IgA and IgG ASCAs. CONCLUSION: In the majority of patients ASCAs disappeared during a GFD. In children this disappearance of ASCA positivity was more pronounced. This can be explained by the well-known fact that gut permeability normalizes much better in children than in adults. Also, the adults had higher levels of ASCAs at diagnosis. This was probably because they had been exposed to gluten for longer and therefore had more long-lasting damage. PMID- 16357624 TI - Accuracy of plasma levels of polymorphonuclear elastase as early prognostic marker of acute pancreatitis in routine clinical conditions. AB - OBJECTIVES: The early prognostic evaluation of acute pancreatitis (AP) is a key step in the appropriate management of the disease. Plasma levels of polymorphonuclear elastase have proved to be an accurate early prognostic marker of AP in research conditions. Whether the test remains sufficiently accurate in routine clinical conditions has been questioned. The aim of our study was to evaluate the accuracy of plasma polymorphonuclear-elastase levels for the early prognostic evaluation of AP in the clinical setting. METHODS: A total of 224 consecutive patients with AP admitted to our Gastroenterology Department were included. A blood sample for polymorphonuclear-elastase quantification was obtained from all of them in the first morning of hospital stay, together with samples for routine haematological and biochemical analysis. Blood samples for polymorphonuclear-elastase evaluation were sent to the laboratory and managed there according to routine protocols. AP was classified as mild or severe according to the Atlanta classification, whereas polymorphonuclear-elastase results were kept blind. Results were shown as mean+/-SD and compared using Student's t-test for unrelated samples. The accuracy of the test for the prognostic evaluation of AP was calculated after drawing the corresponding receiver operator curve. RESULTS: Fifty patients (23%) suffered from severe AP. The plasma levels of polymorphonuclear elastase were 217.8+/-93.5 microg/l in patients with severe AP and 68.1+/-32.7 microg/l in those with mild disease (P<0.001). The sensitivity and specificity of the test for the detection of severe AP were 92 and 91%, respectively, for an optimal cut-off value of 110 microg/l. The positive and negative predictive values for a prevalence of severe disease of 20% were 78 and 96%, respectively. The area under the receiver operator curve was 0.956. CONCLUSION: Quantification of plasma polymorphonuclear elastase levels is a very accurate method for the early prognostic evaluation of AP, and is easily applicable in the clinical setting. PMID- 16357625 TI - An office-based serological test for detection of current Helicobacter pylori infection: is it useful? AB - OBJECTIVE: To compare the accuracy of a new office-based serological test for the detection of current Helicobacter pylori infection with gastric histopathology. METHODS: An office-based serological test for detection of current H. pylori infection was performed in 92 consecutive patients with dyspeptic symptoms. The result of this test was compared blindly with the rapid urease test and histopathology for H. pylori on gastric biopsies. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of this test were calculated against the histopathology. RESULTS: Among 92 patients, H. pylori was detected by histopathology and rapid urease test in 61 (66%) and 56 (60%) patients, respectively. Meanwhile the serological test diagnosed current H. pylori infection in 55 patients (59%). On comparison against the histology, the sensitivity and specificity of this new serological test was 85 and 90%, respectively. The positive predictive value and negative predictive value of this test were 94 and 76%, and the accuracy was 87% in this study. CONCLUSION: The specificity and sensitivity of the serological test was found comparable with histopathology in the diagnosis of current H. pylori infection. This test is a useful tool for rapid diagnosis of H. pylori in out patients' setting. PMID- 16357626 TI - Multipolar radiofrequency ablation of large hepatic metastases of endocrine tumours. AB - Radiofrequency ablation (RFA) is a reliable method of creating thermally induced coagulation necrosis. Local recurrence after RFA of hepatic metastases is directly dependent on tumour size related to the free margin of ablation. To produce larger coagulation volumes a bipolar radiofrequency device was developed that allows the simultaneous activation of three active needles. This technique was used at laparotomy in a patient with liver metastases of an endocrine tumour. Coagulation size up to 12 cm in diameter could be created. The postoperative recovery of the patient was uncomplicated. No local recurrence was seen after 13 months of follow-up with computed tomography scan. The use of simultaneously operated multiple radiofrequency electrodes in a multipolar mode expands the treatment options for patients with large and unresectable intrahepatic metastases. PMID- 16357627 TI - Diagnosis and treatment of VIPoma in a female patient. AB - We report a case of VIPoma in an 83-year-old female patient, who presented with frequent and excessive diarrhoea, muscle weakness, and severe hypokalaemia. Abdominal computed tomography (CT) revealed a 4x6 cm mass in the body of the pancreas. Laboratory analysis showed elevated levels of both vasoactive intestinal polypeptide (VIP; 153 pmol/l) and pancreatic polypeptide (161 pmol/l). In view of the patient's age, physical condition, and tumour size, surgical resection was not performed. The patient was treated with a long-acting octreotide, after which her symptoms diminished. After 24 months of follow-up, the patient remained in good physical condition without any further serious gastrointestinal symptoms. The VIPoma syndrome is caused by a neuroendocrine tumour, usually located in the pancreas, which secretes VIP, causing severe diarrhoea, dehydration and hypokalaemia. Treatment options include resection of the tumour, chemotherapy or the reduction of symptoms with somatostatin analogues. We provide an overview of the incidence, pathophysiology, diagnosis, treatment strategies, and prognosis of this rare syndrome. PMID- 16357628 TI - Non-alcoholic fatty liver disease in HIV-positive patients predisposes for acute on-chronic liver failure: two cases. AB - Non-alcoholic fatty liver disease is a prominent feature in HIV-positive patients. We present two patients with long-lasting HIV-infection who suffered from this disease, as induced by highly active anti-retroviral therapy (HAART). The patients developed acute-on-chronic (AOC) liver failure after either (case 1) acute infection with hepatitis A virus (HAV) or (case 2) methamphetamine abuse ('Ecstasy'). Approximately 1 week after visiting an area endemic for HAV, case 1, a male patient, presented with icterus, elevated liver transaminases and HAV IgM. Previous examinations had demonstrated normal liver transaminase activities while hepatic steatosis had been suspected. He developed complications associated with liver failure including renal failure as well as pleural and pericardial effusions. Case 2, a second male patient, developed both liver failure and lactic acidosis 24 h after methamphetamine abuse. Both patients suffered from fatty liver in the pre-acute stage as indicated by ultrasound examination. After developing symptoms of liver failure, HAART was discontinued in both patients. Follow-up visits demonstrated that the patients recovered clinically with almost normalized laboratory parameters. In HIV infection, HAART-induced hepatopathological alterations may exist despite the absence of relevant laboratory parameters. These patients are likely to develop AOC liver failure when subjected to acute risk factors such as hepatitis viruses and narcotics or other drugs. In patients treated with HAART, we thus highly recommend hepatitis A and B virus vaccinations, and close monitoring of liver parameters. PMID- 16357629 TI - Severe acute haemorrhagic gastritis controlled by hydrogen peroxide. AB - A 92-year-old woman presented with severe acute haemorrhagic gastritis due to abuse of non-steroidal anti-inflammatory drugs (NSAIDs). She was treated with instillation of 150 ml 3% hydrogen peroxide (H2O2) every 2 h via a nasogastric tube. The copious amount of bright red blood through the nasogastric tube started to decline substantially after the first administration of H2O2 and continued to reveal clear material during the second and third instillation of H2O2. The total amount of H2O2 administered was 600 ml. No rebleeding and only a few flame-shaped intramucosal haemorrhages were observed on the following four consecutive daily endoscopic evaluations. These are promising observations which will have to be confirmed with respect to the safety and efficacy of H2O2 treatment by further controlled studies. PMID- 16357630 TI - Resolution of a complex mediastinal pseudocyst in a patient with alcohol-related chronic pancreatitis following abstinence from alcohol. AB - We describe a 55-year-old patient with alcoholic chronic pancreatitis, a large mediastinal pseudocyst and a hemorrhagic pleural effusion. A single thoracocentesis and withdrawal of alcohol without other conservative or invasive measures resulted in a complete resolution of the pleural effusion and the mediastinal pseudocyst, which to our knowledge has not been described before. PMID- 16357631 TI - Identifying hereditary non-polyposis colorectal cancer. PMID- 16357632 TI - Proximal splenic artery embolization allows pegylated interferon and ribavirin combination therapy in chronic hepatitis C virus-infected patients with severe cytopenia. PMID- 16357633 TI - Neurocomputational models in the study of epileptic phenomena. PMID- 16357634 TI - Epileptic transitions: model predictions and experimental validation. AB - The essence of epilepsy is that a patient displays (long) periods of normal EEG activity (i.e., nonepileptiform) intermingled occasionally with epileptiform paroxysmal activity. The mechanisms of transition between these two types of activity are not well understood. To provide more insight into the dynamics of the neuronal networks leading to seizure generation, the authors developed a computational model of thalamocortical circuits based on relevant patho(physiologic) data. The model exhibits bistability, i.e., it features two operational states, ictal and interictal, that coexist. The transitions between these two states occur according to a Poisson process. An alternative scenario for transitions can be a random walk of network parameters that ultimately leads to a paroxysmal discharge. Predictions of bistable computational model with experimental results from different types of epilepsy are compared. PMID- 16357635 TI - Understanding the transition to seizure by modeling the epileptiform activity of general anesthetic agents. AB - A central difficulty in modeling epileptogenesis using biologically plausible computational and mathematical models is not the production of activity characteristic of a seizure, but rather producing it in response to specific and quantifiable physiologic change or pathologic abnormality. This is particularly problematic when it is considered that the pathophysiological genesis of most epilepsies is largely unknown. However, several volatile general anesthetic agents, whose principle targets of action are quantifiably well characterized, are also known to be proconvulsant. The authors describe recent approaches to theoretically describing the electroencephalographic effects of volatile general anesthetic agents that may be able to provide important insights into the physiologic mechanisms that underpin seizure initiation. PMID- 16357636 TI - EEG analysis with nonlinear excitable media. AB - The detection of patterns embedded within a complex, nonstationary, and noisy background activity is a crucial and important task in EEG analysis. The authors present a biologically inspired, analog approach to EEG analysis that is conceptually different from a variety of statistical approaches currently used. A nonlinear, excitable, spatially extended medium that is composed of diffusively coupled model neurons is considered. When EEG recordings are applied as local perturbations to such an excitable neural tissue, the induced transient changes in the dynamics of the perturbed system can be regarded as an instantaneous characterization of transient processes in the brain reflected by the EEG, e.g., in the form of a sequence of correlated dynamical events (patterns). Nonlinear excitable media can be implemented in form of an array of locally coupled integrated analog nonlinear electrical circuits called cellular neural networks, which represent a next evolutionary step in the parallel analog computer architecture. Using cellular neural networks, the authors show that the concept of signal-induced pattern generation allows an almost instantaneous and unsupervised detection of seizure onsets in EEG recordings. In addition, they show that a cellular neural network can be trained in a supervised way to approximate the degree of synchronization in EEG recordings. The resulting pattern-recognition device may be suitable for the prediction of epileptic seizures. PMID- 16357637 TI - Combined experimental/simulation studies of cellular and network mechanisms of epileptogenesis in vitro and in vivo. AB - The electrical correlates of clinical seizures, and of experimental models of seizures, are recognized because neurons behave differently than normal. Individual neurons do unusual things, and neuronal activities become correlated with each other in ways that are not observed under physiologic conditions. Single neurons may fire bursts of action potentials superimposed on large depolarizations, and the bursts may recur rhythmically over a wide range of frequencies (<1 Hz to 25 Hz); periods of noisy subthreshold activity can occur; and firing can even be suppressed in some neurons. At the population level, nearby neurons tend to fire action potentials, or generate bursts, that are temporally locked together on a few-milliseconds time scale, so that large voltage transients are generated in extracellular fields. Very fast oscillations (>80 Hz) in neuronal aggregates may occur before, during, or after such large field potentials. Finally, cellular activities may even be correlated across large brain regions. The authors review some of the means by which cellular intrinsic properties, synaptic interactions, and electrical coupling via gap junctions, all contribute to the anomalous population activities characteristic of seizures. Also reviewed are some of the data suggesting that the requisite gap junctions are located on the axons of glutamatergic neurons. PMID- 16357639 TI - Alive and very well: with appreciation. PMID- 16357638 TI - Interictal to ictal transition in human temporal lobe epilepsy: insights from a computational model of intracerebral EEG. AB - In human partial epilepsies and in experimental models of chronic and/or acute epilepsy, the role of inhibition and the relationship between the inhibition and excitation and epileptogenesis has long been questioned. Besides experimental methods carried out either in vitro (human or animal tissue) or in vivo (animals), pathophysiologic mechanisms can be approached by direct recording of brain electrical activity in human epilepsy. Indeed, in some clinical presurgical investigation methods like stereoelectroencephalography, intracerebral electrodes are used in patients suffering from drug resistant epilepsy to directly record paroxysmal activities with excellent temporal resolution (in the order of 1 millisecond). The study of neurophysiologic mechanisms underlying such depth-EEG activities is crucial to progress in the understanding of the interictal to ictal transition. In this study, the authors relate electrophysiologic patterns typically observed during the transition from interictal to ictal activity in human mesial temporal lobe epilepsy (MTLE) to mechanisms (at a neuronal population level) involved in seizure generation through a computational model of EEG activity. Intracerebral EEG signals recorded from hippocampus in five patients with MTLE during four periods (during interictal activity, just before seizure onset, during seizure onset, and during ictal activity) were used to identify the three main parameters of a model of hippocampus EEG activity (related to excitation, slow dendritic inhibition and fast somatic inhibition). The identification procedure used optimization algorithms to minimize a spectral distance between real and simulated signals. Results demonstrated that the model generates very realistic signals for automatically identified parameters. They also showed that the transition from interictal to ictal activity cannot be simply explained by an increase in excitation and a decrease in inhibition but rather by time-varying ensemble interactions between pyramidal cells and local interneurons projecting to either their dendritic or perisomatic region (with slow and fast GABAA kinetics). Particularly, during preonset activity, an increasing dendritic GABAergic inhibition compensates a gradually increasing excitation up to a brutal drop at seizure onset when faster oscillations (beta and low gamma band, 15 to 40 Hz) are observed. These faster oscillations are then explained by the model feedback loop between pyramidal cells and interneurons targeting their perisomatic region. These findings obtained from model identification in human temporal lobe epilepsy are in agreement with some results obtained experimentally, either on animal models of epilepsy or on the human epileptic tissue. PMID- 16357641 TI - How did our Academy Journal start and become preeminent? PMID- 16357643 TI - Investigation of the performance of the Menifocal Z gas-permeable bifocal contact lens during continuous wear. AB - PURPOSE: The Menifocal Z is an alternating vision, concentric, bifocal gas permeable (GP) contact lens; center distance is connected to near periphery by a smooth transition zone. The lens is produced using tisilfocon A (Menicon Z material), which is approved for up to 30 days of continuous wear (CW). The aim of this study was to evaluate the clinical performance of the Menifocal Z when worn for up to 30 days of CW for 6 months. METHODS: Thirty-five existing GP lens wearers were enrolled in the study. Subjects were fitted with Menifocal Z lenses and follow-up visits were conducted after 2 weeks of daily wear and 1 day, 1 week, 6 weeks, 3 and 6 months of CW. A range of objective and subjective clinical performance measures were assessed, including distance and near visual acuity, the physiological response to CW, and subjective evaluation of vision and comfort. RESULTS: Twenty-seven subjects (77%) completed the study and eight (23%) discontinued: five (14%) as a result of lens-related problems (four vision, one comfort) and three (9%) as a result of non-lens related reasons. Average CW time achieved by the subjects was 22 +/- 2 days. Mean binocular logarithm of the minimum angle of resolution (logMAR) acuities at 6 months were: high contrast distance 0.03 (20/20-), low contrast distance 0.63 (20/80-), and high contrast near 0.26 (20/25, N4). Adverse responses and lens binding were minimal, and there were no significant increases in corneal staining, corneal vascularization, or superior palpebral conjunctival papillae over time (p > 0.05). Problems with night vision (distance and near) with the lenses were the most common difficulties reported by the subjects. CONCLUSIONS: The Menifocal Z appears to be a promising option for presbyopic vision correction, providing successful correction of distance and near vision in a group of experienced GP lens wearers. The hyper Dk tisilfocon A (Menicon Z) material allowed for safe wear of the lenses on a CW basis. PMID- 16357642 TI - Comparison of flat and steep rigid contact lens fitting methods in keratoconus. AB - PURPOSE: The purpose of this article is to compare the safety and efficacy of flat- and steep-fitting rigid contact lenses in keratoconus. METHODS: The Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study is a 16-center observational study. Cross-sectional results at baseline were generated for 1091 subjects with longitudinal results from the 871 subjects who completed 8 years of follow up. RESULTS: Of the 761 rigid contact lens-wearing patients at baseline, 41% had a scar at baseline compared with 24% of the nonrigid contact lens wearers (odds ratio [OR], 2.15; 95% confidence interval [CI], 1.35-3.43; p = 0.001). Eighty-seven percent were fitted with flat-fitting lenses, whereas 13% were fitted with steep-fitting lenses. Rigid lens fitting method was also associated with incident corneal scarring. A greater proportion of the corneas wearing flat fitting contact lenses were scarred (43% compared with 26% for the steep-fitted eyes; OR,= 2.19; 95% CI, 1.37-3.51; p = 0.001). After controlling for corneal curvature, the association of rigid contact lens fit and corneal scarring at baseline did not persist (adjusted OR, 1.20; 95% CI, 0.70-2.06; p = 0.52). Thirty two percent of unscarred eyes at baseline fitted flat had developed an incident corneal scar by the eighth year follow-up visit compared with 14% of eyes fitted steep (OR, 2.93; 95% CI, 1.34-6.42; p = 0.007). CONCLUSIONS: The data reported here indicate that, after controlling for disease severity in the form of corneal curvature, keratoconic eyes fitted with a rigid contact lens resulting in an apical touch fluorescein pattern did not have an increased risk of being scarred centrally at baseline. This "natural history" sample cannot determine causal proof that one method of fitting lenses is safer than another. To achieve this, a randomized clinical trial is needed. PMID- 16357644 TI - The agreement and repeatability of tear meniscus height measurement methods. AB - PURPOSE: This study investigated the agreement and repeatability of five techniques of measuring tear meniscus height (TMH). METHODS: Pairs of TMH measurements were obtained from one eye of 25 normal subjects with five techniques: optical coherence tomography in cross-section (OCT) and optical pachymetry in cross-section (PACH), without fluorescein; and 5 minutes after the instillation of fluorescein, optical pachymetry en-face (SL-F) and in cross section (SL-X), and video capture en-face (VC). These 10 measurements were taken on different days within a period of 21 days. Additionally, immediately successive replicates were taken at the first visit when VC was used, allowing for the separation of technique repeatability from true day-to-day variation in TMH. RESULTS: Ninety-five percent confidence intervals for the mean TMH and 95% repeatability coefficients for repeated testing on separate occasions were: 0.25 to 0.29 mm, 0.14 mm--OCT, 0.30 to 0.36 mm, 0.13 mm--PACH, 0.27 to 0.34 mm, 0.092 mm--SL-F, 0.35 to 0.41 mm, 0.19 mm--SL-X, and 0.30 to 0.38 mm, 0.088 mm--VC. Reported repeatability coefficients are averages because, despite a relatively narrow experimental range, the measurements of low TMH were generally more repeatable than those of large TMH. Ninety-five percent limits of agreement were wide for all pairwise comparisons and included zero, indicating that the methods used in this study are interchangeable when taking single recordings, at least on subjects without dry eye. CONCLUSIONS: The average TMH of the five techniques was similar, although OCT and SL-X tended to record lower and higher values, respectively. However, their agreement for single recordings was poor, an inevitable consequence of the low intertest repeatability found for all methods. A much improved repeatability for immediate replicates indicated that this, in turn, was principally caused by a large day-to-day variation in TMH rather than measurement error. PMID- 16357645 TI - Automated decision tree classification of corneal shape. AB - PURPOSE: The volume and complexity of data produced during videokeratography examinations present a challenge of interpretation. As a consequence, results are often analyzed qualitatively by subjective pattern recognition or reduced to comparisons of summary indices. We describe the application of decision tree induction, an automated machine learning classification method, to discriminate between normal and keratoconic corneal shapes in an objective and quantitative way. We then compared this method with other known classification methods. METHODS: The corneal surface was modeled with a seventh-order Zernike polynomial for 132 normal eyes of 92 subjects and 112 eyes of 71 subjects diagnosed with keratoconus. A decision tree classifier was induced using the C4.5 algorithm, and its classification performance was compared with the modified Rabinowitz McDonnell index, Schwiegerling's Z3 index (Z3), Keratoconus Prediction Index (KPI), KISA%, and Cone Location and Magnitude Index using recommended classification thresholds for each method. We also evaluated the area under the receiver operator characteristic (ROC) curve for each classification method. RESULTS: Our decision tree classifier performed equal to or better than the other classifiers tested: accuracy was 92% and the area under the ROC curve was 0.97. Our decision tree classifier reduced the information needed to distinguish between normal and keratoconus eyes using four of 36 Zernike polynomial coefficients. The four surface features selected as classification attributes by the decision tree method were inferior elevation, greater sagittal depth, oblique toricity, and trefoil. CONCLUSION: Automated decision tree classification of corneal shape through Zernike polynomials is an accurate quantitative method of classification that is interpretable and can be generated from any instrument platform capable of raw elevation data output. This method of pattern classification is extendable to other classification problems. PMID- 16357646 TI - Orbscan global pachymetry: analysis of repeated measures. AB - PURPOSE: The Orbscan II is a hybrid slit-scanning and Placido disc corneal topographer capable mapping global pachymetry over a 10-mm corneal diameter. In this study, the repeatability of the Orbscan global pachymetry was determined. METHODS: Five consecutive Orbscan examinations were performed on one eye of 20 healthy volunteers by one examiner in one session. Mean, standard deviation, range of readings, and coefficient of variance were calculated for the central, thinnest point and global pachymetry. RESULTS: Orbscan repeatability for the central pachymetry was 3.62 microm (95% confidence interval [CI], 0.57-6.66) with a coefficient of variance of 0.67% (95% CI, 0.09-1.25) and intraclass correlation coefficient (ICC) of 0.978 (95% CI, 0.959-0.990). Thinnest point pachymetry had a repeatability of 4.25 microm (95% CI, 1.19-7.31) with a coefficient of variance of 0.80% (95% CI, 0.19-1.42) and ICC of 0.973 (95% CI, 0.950-0.988). Global pachymetry had a coefficient of variance <1.5% in the central 4 mm and <<2.5% across the entire cornea. Global pachymetry showed an area of greatest repeatability (<< +/- 5 microm) within a central 3.0-mm horizontal and 4.0-mm vertical diameter. DISCUSSION: Orbscan is capable of giving repeatable pachymetry readings that are comparable to the ultrasound pachymeter within the central and thinnest point of the cornea. There is a gradual loss of repeatability toward the peripheral cornea possibly as a result of a lack of overlapping data points. PMID- 16357647 TI - The reliability of the Lanthony Desaturated D-15 test. AB - PURPOSE: The Lanthony Desaturated Panel D-15 has been used to measure fine color discrimination for congenital and acquired color vision defects. This study investigated the test-retest reliability of the test using an intertest interval of approximately 1 month. METHODS: One hundred twenty-six color vision normals (mean age = 34.5 years) were administered several color vision tests, including the Lanthony Desaturated D-15. Normal color vision status was confirmed using the anomaloscope and HRR color plates. The color vision tests were readministered 3 to 6 weeks after initial testing. The results of the Lanthony test were expressed using the color confusion index of Bowman. The difference in Color Confusion Index (CCI) between the two administrations was calculated and used to determine the intraclass correlation coefficient. RESULTS: The overall mean CCI for the two administrations for these subjects was 1.11 +/- 0.136. The mean difference in CCI score between test administrations was -0.02 +/- 0.128. There was a strong correlation between the absolute value of the difference in CCI and the mean CCI for each subject (r = 0.51, p < 0.0001). The intraclass correlation coefficient was 0.56 (95% confidence interval, 0.43-0.67). CONCLUSIONS: Although the Lanthony Desaturated D-15 test can be used to assess fine color discrimination, there is considerable within-subject variability in test results. The intraclass correlation coefficient is less than that recommended for use in clinical testing or research. Clinicians should consider at least three administrations of the test at each sitting to ensure precision and we recommend taking the mean of those three tests. PMID- 16357648 TI - Dynamics of saccadic adaptation: differences between athletes and nonathletes. AB - PURPOSE: The aim of the study was to delineate differences in saccadic adaptation characteristics between a population of racquet sports athletes and nonathletes. METHODS: Eye movements were recorded at 120 Hz using a video-based eye tracker (ELMAR 2020) in a sample of 27 athletes (varsity badminton and squash players) and 14 nonathletes (<3 hours/week participation in recreational sports). Responses to negative positional error and positive positional error were studied in two sessions on separate days. Negative positional errors were induced by displacing the stimuli backwards by 3 degrees from the initial target step (12 degrees). Likewise, positive positional errors were induced by displacing the stimuli forward by 3 degrees . Amplitude gains were calculated for trials before, during, and after the adaptation phase. The magnitude and the rate of change of saccadic adaptation were determined from the amplitude gains. Differences between the groups were compared using regression analysis. RESULTS: No significant differences were found between the two groups in the magnitude of saccadic adaptation, both for negative (athletes -60%, nonathletes -57%) and positive (athletes +26%, and nonathletes +27%) positional error. Racquet sports athletes showed a significantly faster rate of adaptation for the positive positional error. A significant difference was not observed in the rate of adaptation for the negative positional error. CONCLUSIONS: Racquet sports athletes and nonathletes adapt to positional error signals by similar amounts. However, racquet sports athletes respond to positive positional errors at a faster rate, suggesting that a strategic component or environmental influences (such as practice) may play a role in saccadic adaptation. PMID- 16357649 TI - Comparisons of the handheld autorefractor, table-mounted autorefractor, and subjective refraction in Singapore adults. AB - PURPOSE: The purpose of this article was to compare the Retinomax with the table mounted autorefractor and subjective refraction in Singapore adults. METHODS: Adults (n = 100) attending a tertiary eye hospital clinic were examined by an optometrist. First, subjective refraction testing was performed using a trial lens set, followed by handheld autorefractor tests using the Nikon Retinomax and the table-mounted autorefractor (Topcon RM8000B). Spherical equivalent and vector components of astigmatism were analyzed: J0 (Cartesian astigmatism) and J45 (oblique astigmatism). RESULTS: The Retinomax autorefractor readings (mean = 4.69 D) were more minus compared with the table-mounted autorefractor (mean = 4.05 D) and subjective refraction (mean = -3.90 D). There were significant differences in J0 and J45 for comparisons between subjective refraction and Retinomax autorefraction, and table-mounted autorefraction and Retinomax autorefraction. CONCLUSION: The Retinomax autorefractor measures were more minus compared with the table-mounted autorefractor and subjective refraction. The Retinomax autorefractor is not recommended for research purposes, unless in remote inaccessible areas where a portable instrument is necessary and cycloplegia is not possible. PMID- 16357650 TI - Presbyopia compensation with a quartic axicon. AB - PURPOSE: The purpose of this study was to evaluate the performance of quartic axicons for presbyopia compensation. The working principle relies on profiting the high depth of focus of the axicons to supplement the reduced accommodation amplitude of presbyopes. METHODS: We present the design equations of a particular kind of axicon to compensate a general presbyopia condition using simultaneous vision. A rotationally symmetric polynomial of fourth-order, corresponding to the well-known Seidel spherical aberration term, was chosen as its refractive profile. To validate its performance, we computed the retinal images with Stiles Crawford apodization for a presbyopic eye compensated with this quartic axicon and compared them with those obtained without compensation or with other available solutions based on the simultaneous vision principle. RESULTS: The quartic axicon provides an important improvement of the image quality for intermediate distance vision in comparison with conventional bifocal and trifocal solutions. The image quality, however, is still not optimum for all distances. CONCLUSIONS: The results show the usefulness of the proposed approach and point out the need for developing further adapted optimizations. PMID- 16357651 TI - The challenge to follow up. PMID- 16357652 TI - Trunk position in supine of infants born preterm and at term: an assessment using a computerized pressure mat. AB - PURPOSE: Trunk position may influence motor, cognitive, and social development during infancy and early childhood and has not been quantitatively assessed. The purpose of this study was to assess the trunk positions of infants born at term and preterm in supine using a computerized pressure mat. METHODS: Trunk position was represented as a ratio of head and pelvis to trunk pressure. Eighteen healthy infants born preterm with a mean gestational age of 31.9 weeks (25.0-34.6) and fifteen healthy infants born at term with a mean gestational age of 38.9 weeks (37.3-40.6) were assessed at 38 to 43 weeks gestational age. RESULTS: Infants born at term spent more than two-thirds of the time in either flexed or neutral trunk positions. No significant differences were found between infants born preterm and those born at term in total duration of flexion or extension or in flexion event duration. CONCLUSION: This study provides evidence that infants born preterm may not exhibit greater trunk extension tendencies in supine than infants born at term. Results should be interpreted with caution, however, because of the small sample size and large variability observed within the subject groups. Infants born at less than 30 weeks of gestation may demonstrate greater extension tendencies than those born at more than 30 weeks of gestation. PMID- 16357653 TI - Repeatability of peak oxygen uptake in children who are healthy. AB - PURPOSE: This study was designed to determine test-retest repeatability of variables during a maximal cardiopulmonary exercise test (CPET) in children, and to report repeatability coefficients using limits of agreement analysis. This method of analysis is recommended for valid interpretation of outcome measures used in clinical or research interventions, and has not been reported in previous studies of peak oxygen uptake (VO2peak) in children. METHODS: Two exercise tests were performed three to seven days apart in nine healthy children aged eight to 11 years, using a treadmill protocol. Limits of agreement analyses were performed, with bias (mean difference test 2 - test 1) and coefficients of repeatability (2 standard deviations [SD] of difference) calculated for VO2peak, peak heart rate (HRpeak) and peak minute ventilation (VEpeak). RESULTS: No significant differences were found between test and retest scores for any variable. VO2peak data demonstrated a bias of 1.4 ml kg/min with repeatability coefficient of 4.4 ml kg/min. The range described by 95% limits of agreement for VO2peak was -3 to + 5 ml kg/min. Bias and 95% limits of agreement for VEpeak and HRpeak were similarly described. CONCLUSION: The 95% limits of agreement for VO2peak imply that on repeated testing, a reduction of 3 or increase of 5.8 ml kg/min is likely to be clinically significant. This study provides reference values for repeatability of key variables during CPET in healthy children that should be considered when interpreting results from repeated testing. PMID- 16357654 TI - Motor and postural response profiles of four children with developmental coordination disorder. AB - PURPOSE: The purpose of this report is to present a descriptive profile of the postural responses and motor skill performance of four children with developmental coordination disorder (DCD). METHODS: Four children with DCD, aged nine to 12 years participated. A profile of each child's case background, school performance, and motor skill performance was developed, in addition to the collection of data from three sessions of posturography trials on the computerized Sensory Organization Test (SOT). RESULTS: All four children had a significant degree of impairment in balance responses and limitations in motor performance, with increased sway and frequency of falls in all sensory conditions on the SOT in comparison with peers. Mean equilibrium scores were between one and two standard deviations below peer averages, and sensory scores showed performance levels similar to those of younger age groups. There was a high level of variability associated with repeated SOTs. CONCLUSIONS: The SOT data provide further insight into the underlying mechanisms that contribute to balance deficits experienced by children with DCD and suggest that assessment and therapeutic approaches take into account sensory deficits in postural control. PMID- 16357655 TI - Effects of aerobic conditioning and strength training on a child with Down syndrome: a case study. AB - PURPOSE: To determine the effects of a home exercise program of combined aerobic and strength training on fitness with a 10.5-year-old girl with Down syndrome (DS). Measurements included cardiovascular variables, strength, body composition, flexibility, and skill. METHODS: The subject participated in a home exercise program: 30 to 60 minutes of moderate- to high-intensity exercise five to six days per week for six weeks. The cardiovascular variables monitored were heart rate, respiration rate, and oxygen consumption during a submaximal treadmill stress test. Other measures included 10-repetition maximal strength of selected muscle groups, body mass index, flexibility, Gross Motor Scales of the Bruininks Oseretsky Test of Motor Proficiency, and anaerobic muscle power. RESULTS: Improvements in submaximal heart and respiration rates, aerobic performance, muscle strength and endurance, gross motor skills, and anaerobic power were observed for this subject. Body weight and flexibility were unchanged. CONCLUSIONS: For this subject, a combined aerobic and strength-training program resulted in improved cardiopulmonary functions not observed in previous studies of subjects with DS. PMID- 16357656 TI - Physical activity monitoring in children and youths. AB - BACKGROUND AND PURPOSE: Public health data have confirmed epidemic levels of pediatric obesity and inactivity in the United States today. Activity monitoring can provide real-world outcomes for new interventions for youths with mobility limitations. Valid and reliable measures of ambulatory activity are needed to understand the relationship between inactivity and obesity. The purpose of this paper is to highlight issues relevant to ambulatory activity monitoring in pediatric populations. SUMMARY OF KEY POINTS: A review of the currently available pedometers and accelerometers for ambulatory physical activity monitoring is presented. Elements of device selection are discussed and critical criteria proposed for physical activity monitoring of children and adolescents. Finally, examples of pediatric physical activity monitoring are presented in the context of clinical practice and research. PMID- 16357657 TI - Exploring physical therapy clinical decision making for children with spastic diplegia: survey of pediatric practice. AB - PURPOSE: The purpose of this special interest report is to describe the outcomes of a research round table discussion regarding the physical therapy management of mobility for children with spastic diplegia. DESCRIPTION: Sixty-two pediatric physical therapists and physical therapists assistants participated in focus groups during the Research Round Table at the American Physical Therapy Association (APTA) 1999 Combined Sections Meeting. A case description of a child with spastic diplegia and guiding questions were used to facilitate discussion. SUMMARY OF EXPERIENCE: Common practices in patient management across the child's life-span emerged from the discussion. Practices in examination, evaluation and prognosis, and intervention differed depending on the age and function of the child and the family's needs. In general, therapists reported that younger children receive examinations that include standardized tests of development and ongoing intervention with a frequency of one to five times per week. In contrast, older children receive therapy services on an episodic basis that address their specific needs. The elements of patient management served as a useful framework for exploring decision making. IMPORTANCE TO PEDIATRIC PHYSICAL THERAPY: The information compiled from this project needs to be validated through systematic inquiry. Therapists may, however, use the practices reported here to reflect on their clinical decision making and to identify questions for further exploration. This descriptive document is the first step in the development of a guideline for evidence-based practice. The development of such a clinical guideline could serve as an education tool for novice therapists, a program evaluation tool to ensure quality care, and a foundation for future research to promote evidence-based practice. PMID- 16357660 TI - Another STEP. PMID- 16357661 TI - The effects of an early physical therapy intervention for very preterm, very low birth weight infants: a randomized controlled clinical trial. AB - PURPOSE: A randomized controlled clinical trial was used to investigate effects of physical therapy (PT) intervention on motor outcome of infants born very preterm with very low birth weight (VLBW). METHODS: Seventy-two infants born very preterm with VLBW were randomly assigned to a nontreatment (NT) (n = 38) or treatment (T) (n = 34) group. The T group received developmental PT from birth until four months corrected age (CA) weekdays during the infant's neonatal stay and on a needs- and problem-orientated basis thereafter. The NT group received no intervention. Both groups were assessed at four months CA using the Alberta Infant Motor Scale (AIMS) as was a control group of 14 infants born full term. Parental compliance was measured using a parent questionnaire. RESULTS: PT intervention had no significant effect on the T group's motor performance. However, no T group subjects had abnormal motor development at four months CA when compared to the NT group (16%) and the control group (14%) (p = 0.09). The T group subjects with high levels of parental compliance had better scores on the AIMS than those with lower parental compliance (p = 0.05). CONCLUSION: PT intervention does not significantly affect motor performance of infants born very preterm with VLBW at four months CA. Parental compliance and intervention frequency may have influenced the outcome. Preliminary evidence suggests that neonatal and early PT may reduce the incidence of motor delay among infants born very preterm with VLBW. Follow-up of this group is recommended to ascertain the long-term benefits of this type of early PT. PMID- 16357662 TI - Expectations for walking speeds: standards for students in elementary schools. AB - PURPOSE: The aim of this study was to determine children's walking speeds in elementary school hallways. A second aim was to determine the slowest but "good enough" walking speed if a teacher had to slow down her class's line to accommodate a student with a mobility limitation. METHODS: The walking speeds of 370 children serving as class-line leaders were determined after measuring using a stopwatch the time that it took to walk along 50-ft paths in school hallways. Teachers were asked to demonstrate a "good enough" speed for a child with mobility limitations after instruction and demonstration of slow walking by a physical therapist (PT). The teachers "good enough" speed was determined after measuring with a stopwatch the time that it took for the teacher to walk a distance of 50 ft imitating a "good enough" speed. RESULTS: Significant speed differences were found between children in kindergarten through sixth grade. The mean speed to walk 50 ft used by kindergarten line leaders was 13.5 seconds, and the speed increased to 50 ft in 10.6 seconds for sixth grade lines. Teacher "good enough" times were significantly different from the children's times at every grade level. CONCLUSIONS: These peer-based school standards can be used by PTs, parents, teachers, and other Individualized Education Program team members to make realistic decisions about future goals for preschool and elementary school children with mobility limitations so that they can participate with their peers. PMID- 16357663 TI - The emerging role of the pediatric physical therapist in evaluation and intervention for individuals with lysosomal storage diseases. AB - PURPOSE: The purposes of this article are to describe the pathology, medical implications, and typical impairments of individuals with various lysosomal storage diseases (LSDs), summarize results of recent clinical trials on medical interventions relevant to physical therapy practice, report new advances in functional measurement, and suggest a framework for physical therapy management and intervention. SUMMARY OF KEY POINTS: Medical and surgical interventions are enabling individuals with LSDs to not only survive but to improve their daily functioning and quality of life. This is likely to become an increasing area of emphasis in pediatric physical therapy, as the intervention emphasis for some individuals will shift from maintenance to restorative programs. RECOMMENDATIONS: We recommend that pediatric physical therapists become familiar with new LSD therapeutics, play a major role in evaluating impairment and functional limitation changes in individuals with LSDs, and become knowledgeable about the indications and precautions for restorative physical therapy programs. PMID- 16357664 TI - Psychometric properties and standardization samples of four screening tests for infants and young children: a review. AB - PURPOSE: This article compares traditional psychometric properties (interrater and test-retest reliability, concurrent and predictive validity), clinical epidemiological characteristics (sensitivity, specificity, and positive predictive values), and standardization samples of four tests useful to pediatric therapists in screening infants and young children for developmental delays. SUMMARY OF KEY POINTS: Pediatric therapists are often involved in screening infants and young children for developmental delay. Ideally, they will use standardized tests that have strong psychometric properties (eg, reliability, validity, sensitivity, specificity). The four tests described in this article vary in meeting these criteria. They vary as well in the domains assessed, age ranges for which they are intended, and desired qualifications of the examiners. CONCLUSIONS: Each of the four tests reviewed has identified strengths and weaknesses. Practicing clinicians should select screening tests based on the test's stated purpose, qualifications of the examiner, age range covered, administration and scoring time, developmental domains encompassed, comparability of the standardization sample, and strength of the test's psychometric properties. PMID- 16357665 TI - Wellness promotion beliefs and practices of pediatric physical therapists. AB - PURPOSE: The purpose of this study was to elucidate wellness promotion beliefs and practices of pediatric physical therapists. METHODS: From a random sample of 500 American Physical Therapy Association Pediatric Section active members, 257 physical therapists (51%) returned usable questionnaires designed to gather information on professional and personal wellness beliefs and practices. RESULTS: Descriptive statistics and chi-square analyses were performed to describe current wellness promotion practices. Most participants considered wellness multidimensional, valued wellness, and incorporated wellness practices into their personal life. Only 54.5% of respondents, however, reported incorporating wellness into pediatric physical therapy practice. A third of the respondents identified themselves as either thinking or preparing to incorporate wellness promotion into practice. Factors associated with wellness promotion were older age group, knowledge, belief that wellness promotion was a physical therapy responsibility, and participation in personal wellness lifestyle activities. The most frequent barriers cited were external factors related to resources, time, and the child/family. CONCLUSIONS: Current pediatric physical therapy practice reflects a more traditional model of care rather than a wellness promotion approach. With a paradigm shift in healthcare toward wellness promotion, pediatric physical therapists need to align practice with current societal needs and national healthcare campaigns. Continuing education programs that are participatory and well matched to the characteristics and needs of the attendees combined with collegial support may prove fruitful in providing pediatric physical therapists with the knowledge, motivation, and strategies needed to accomplish this goal. PMID- 16357666 TI - Effects of facilitated tucking during routine care of infants born preterm. AB - PURPOSE: The purpose of this study was to compare stress responses of infants born preterm during routine nursing assessments performed under two conditions. One condition incorporated a second caregiver supporting the infant in a facilitated tucked position, whereas the second condition did not. METHODS: A convenience sample of 12 infants born preterm, ages 25 to 34 weeks postconceptual age on the day of testing (mean = 30.9 weeks), were evaluated using the Premature Infant Pain Profile (PIPP), during the two caregiving conditions. For each trial, the infant received a PIPP score. The level of significance was set at p = 0.05. RESULTS: A significant difference (p = 0.013) existed between the two testing positions as measured by the PIPP. Nine of the 12 infants received a lower PIPP score with facilitated tucking during routine care assessments. CONCLUSIONS: By incorporating facilitated tucking during routine care events, the stress level of the infants born preterm may be reduced. When the infants' stress levels are reduced, they may be better able to maintain stability in their autonomic, motor, and state systems. PMID- 16357669 TI - Collaboration. PMID- 16357671 TI - Therapists' communication styles and parents' global trust in the therapists: a comparison between Jewish and Bedouin parents. AB - PURPOSE: This study was designed to examine the level of global trust in therapists for two groups of Israeli parents, Jews and Bedouin, and to assess which of three interpersonal communication styles significantly explain global trust in the therapists. METHODS: A total of 193 parents of children ranging from six months to six years of age participated in this study. Parents' perceptions regarding their communication with the center's therapists were measured by 15 items clustered into three dimensions of communication: caring, collaboration, and interest. RESULTS: Parents of both cultural groups expressed high levels of global trust in the center's therapists. Jewish parents trusted their children's therapists significantly more than their Bedouin counterparts and ranked the therapists' caring and collaboration more highly. The communication style of caring was found to be the sole contributor to the variability in global trust. CONCLUSIONS: Effort should be undertaken to develop effective communication styles for the dimension of caring to increase therapists' ability to promote global trust in their patients' parents. PMID- 16357672 TI - Outcome of an exercise and educational intervention for children who are overweight. AB - PURPOSE: The purpose of this study was to investigate the outcome of an education and exercise-based intervention for children who are overweight. METHODS: Forty one children with a mean age of 10.5 years participated in the study. The eight week intervention, scheduled twice weekly, included education and exercise components. Outcome measures included body mass index (BMI), waist and hip girth, blood pressure (BP), resting heart rate (RHR), immediate postexercise heart rate (HRfinish), five-minute recovery heart rate (5minHR), and distance walked in six minutes. Using a repeated-measures design, data were collected twice prior to intervention to determine baseline and once upon completion of the intervention. RESULTS: No significant difference existed between pretest I and II measures for each dependent variable, indicating a stable baseline. An intervention effect was established as evidenced by significant improvement in BMI, waist and hip girth, BP, RHR, HRfinish, and 5minHR at posttest. CONCLUSIONS: Children who are overweight and participate in an eight week exercise and educational program demonstrate improved morphology and physical conditioning. PMID- 16357673 TI - The influence of concurrent cognitive tasks on postural sway in children. AB - PURPOSE: The purpose of this study was to examine the influence of concurrent tasks on postural sway in children. METHODS: Nineteen fourth-grade students, while standing on a balance platform, were asked to stand still, count backward, and read second-grade level sentences. The AMTI Accusway System was used to calculate the length of center of pressure path (LCOP), sway range (SR), and variability (SV) in mediolateral (ML) and anteroposterior (AP) directions of sway. RESULTS: Analysis of variance revealed a main effect of cognitive task condition for SR-AP, SR-ML, SV-AP, and SV-ML. Post hoc comparisons revealed lower values of those four dependent measures for the counting backward task than for the standing still task and lower SV-AP for the counting backward task than for the reading task. In addition, there was a trend toward greater LCOP when performing a concurrent cognitive task. CONCLUSIONS: The demands of concurrent cognitive tasks while standing affect postural sway in children. The findings of this study contribute to our understanding of postural control in children and may explain why improvements in postural skills attained in clinical settings may not transfer to improved performance in other settings. PMID- 16357674 TI - Effectiveness of developmental intervention in the neonatal intensive care unit: implications for neonatal physical therapy. AB - PURPOSE: Interdisciplinary team members interact with infants to facilitate progressive physiologic stability. The focus of the physical therapist's role is promotion of sensorimotor development in infants born preterm. The aim of this review was to examine evidence for physical therapist practice in the neonatal intensive care unit (NICU) as it relates to developmental intervention (DI) for infants born prematurely and to present the evidence of physical therapy techniques used in the NICU. SUMMARY OF KEY POINTS: A literature review was performed resulting in identification of 26 articles that examined specific developmental intervention techniques. The articles were critiqued based on their design. Twelve articles were rated highly, indicating that sensory techniques implemented by physical therapists appear to be an appropriate and effective component of DI. The general consensus was that there is a lack of large, well controlled, randomized studies in this area of pediatric outcome research. CONCLUSIONS: Neonatal physical therapy falls under the umbrella of DI. There is substantial agreement about the benefits of DI, but the multimodal and interdisciplinary nature of the evidence limits the ability to identify the effectiveness of any one healthcare professional in the provision of DI in the NICU. PMID- 16357675 TI - TAMO therapy as a major component of physical therapy intervention for an infant with congenital muscular torticollis: a case report. AB - PURPOSE: Tscharnuter Akademie for Motor Organization (TAMO) therapy is a therapeutic approach based on dynamic theories of motor control. Research evidence supporting this approach is limited. The purpose of this case report is to describe the use of TAMO therapy in the physical therapy intervention for an infant with congenital muscular torticollis (CMT). The patient was a 4.5-month old baby boy (corrected age) with left CMT. METHODS: Intervention included TAMO therapy, active range of motion exercise, soft tissue mobilization, and parent instruction. Changes in the amount of lateral head tilt were documented using still photography RESULTS: The infant assumed a midline head position in the supine position by the second weekly visit and maintained a midline head alignment during all functional activities by the eighth visit. CONCLUSION: This case report is the first attempt to describe a successful application of TAMO therapy as a major component of physical therapy intervention for an infant with CMT. PMID- 16357676 TI - Responsiveness of the Test of Infant Motor Performance (TIMP) in infants born preterm. AB - PURPOSE: The responsiveness of the Test of Infant Motor Performance (TIMP) was examined in infants born preterm. Infant variables predictive of changes in TIMP scores were also identified. METHODS: Twenty-five infants born 0.75) for elbow flexors and hamstrings and poor interrater reliability (ICC <0.50) for other muscles. Intrarater scores were good (ICC >0.75) for hamstrings and moderate (ICC = 0.50 to 0.75) for other muscles. CONCLUSION: Interrater reliability of the MAS may be lower than desired for clinical use for muscles other than hamstrings and elbow flexors, and intrarater reliability may also be lower than desired for muscles other than the hamstrings. PMID- 16357683 TI - Characteristics of hypotonia in children: a consensus opinion of pediatric occupational and physical therapists. AB - PURPOSE: The term hypotonia is often used to describe children with reduced muscle tone, yet it remains abstract and undefined. The purpose of this study was to identify characteristics of children with hypotonia to begin the process of developing an operational definition of hypotonia. METHODS: Three hundred physical and occupational therapists were systematically selected from the memberships of the Pediatric Section of the American Physical Therapy Association and the Developmental Delay Section of the American Occupational Therapy Association and asked to complete an open-ended survey exploring characteristics of strength, endurance, mobility, posture, and flexibility. RESULTS: The response rate was 26.6%. Forty-six physical therapists and 34 occupational therapists participated. The criterion for consensus about a characteristic was being mentioned by at least 25% of respondents from each discipline. The consensus was that children with hypotonia have decreased strength, decreased activity tolerance, delayed motor skills development, rounded shoulder posture, with leaning onto supports, hypermobile joints, increased flexibility, and poor attention and motivation. CONCLUSION: An objective tool for defining and quantifying hypotonia does not exist. A preliminary characterization of children with hypotonia was established, but further research is needed to achieve objectivity and clarity. PMID- 16357688 TI - Children are not small adults. PMID- 16357690 TI - Treatment and outcomes of diabetic muscle infarction. AB - BACKGROUND: Diabetic patients can develop spontaneous infarctions of muscle. The optimal treatment of this diabetic muscle infarction (DMI) is unknown. OBJECTIVE: This analysis was conducted to compare the outcomes of conservative, medical, and surgical treatments of DMI. The primary outcome is the time to recovery. Secondary outcomes include recurrence and mortality rates. METHODS: A MEDLINE search from its inception to December 2002 was used to identify reported cases of DMI. We selected those cases that reported on specified baseline characteristics of the patients, including age, gender, duration of diabetes, type of diabetes, diabetic microvascular and macrovascular complications, and the magnetic resonance imaging or computed tomography findings, the type of therapy provided, the time to recovery of initial muscle infarction, recurrences, and deaths. RESULTS: A total of 36 references meeting our inclusion criteria were retrieved, describing 49 patients. Thirty-four patients received conservative therapy (bedrest and analgesics), 8 received medical therapy (antiplatelet agents and/or steroids), and 7 had surgical excision of the infarcted muscle. There were no significant differences in baseline characteristics. The time to recovery from treatment onset was 8.1 weeks, 5.5 weeks, and 13 weeks in the conservative, medical, and surgical treatment groups, respectively. This was statistically significant only when comparing medical and surgical treatment. The respective recurrence rates were 35%, 29%, and 71%. The respective mortality rates within 2 years were 4%, 14%, and 29%. CONCLUSION: This study supports the use of nonsurgical treatment in patients with DMI. It also demonstrates that DMI can be temporally associated with death. PMID- 16357689 TI - Comparison of intravenous pamidronate to standard therapy for osteoporosis: use in patients unable to take oral bisphosphonates. AB - BACKGROUND: Bisphosphonates are potent therapies for osteoporosis, but some patients cannot tolerate oral forms. Our study sought to examine the effectiveness and safety of intravenous pamidronate as an alternate osteoporotic therapy in these patients. METHODS: We conducted a retrospective study of 26 patients treated with intermittent, intravenous pamidronate (30 mg every 3 months) and 52 matched controls treated with standard therapy, all with osteoporosis. Primary outcome was response to therapy, defined as either stabilization or increase in bone mineral density. Secondary outcomes were patient safety and tolerability. RESULTS: At an average of 16.1 months of follow up, 64% of pamidronate-treated patients responded to therapy at the lumbar spine, 65% at the femoral neck, and 63% at the trochanter. These response rates were not significantly different than those of the standard care group in which 69% were receiving oral bisphosphonates. Adverse events were uncommon and included mild, diffuse myalgias and flu-like symptoms. CONCLUSIONS: Intravenous pamidronate is a well-tolerated osteoporosis therapy that has an effect on bone density comparable to standard therapy in patients unable to take oral bisphosphonates. PMID- 16357691 TI - Changes in the prevalence of US adults with arthritis who meet physical activity recommendations, 2001-2003. AB - CONTEXT: Increased physical activity is recommended for adults with arthritis. OBJECTIVE: The objective of this study was to update and compare recent physical activity prevalence estimates of US adults with self-reported doctor-diagnosed arthritis. DESIGN: The authors studied the 2001 and 2003 Behavioral Risk Factor Surveillance Survey (BRFSS), an annual random-digit telephone survey conducted by state health departments. SETTING: The authors conducted a population-based sample from 50 states and the District of Columbia. PARTICIPANTS: The study included a total of 52,311 adults, 18 years or older, reporting doctor-diagnosed arthritis in 2001, and 82,839 reporting doctor-diagnosed arthritis in 2003. MAIN OUTCOME MEASURES: Reported level of physical activity was classified into 3 categories: physically inactive, insufficiently active, and meeting public health recommendations. RESULTS: Between 2001 and 2003, the prevalence of physical inactivity decreased by 1.1% (23.8% versus 22.7%), the prevalence of insufficient levels of physical activity increased by 0.2% (38% versus 38.2%), and the prevalence of meeting physical activity recommendations increased by 0.9% (38.3% versus 39.2%). Compared with 2001 estimates, the prevalence of meeting physical activity recommendations increased by 6.2% (54.8% versus 61%) among those aged 18 to 24 years, 3.5% (36.4% versus 39.9%) among never smokers, 3% (49.4% versus 52.4%) among those aged 25 to 34 years, 2.4% (29.9% versus 32.3%) among obese adults, and among those aged 55 to 64 years (36.9% versus 39.3). Prevalence of inactivity increased by 3.2% among Hispanics (30.4% versus 33.6%) and 2.9% among those with 8 or fewer years of education (47.6% versus 50.5%). CONCLUSIONS: Prevalence estimates of the 3 levels of physical activity among adults with arthritis changed very little. Given the established benefits of physical activity for arthritis, public health efforts to promote increased activity should continue to be developed and refined. PMID- 16357692 TI - Tuberculosis of the spine: experience in an inner city hospital. AB - BACKGROUND: It is estimated that 10 to 15 million people are infected with Mycobacterium tuberculosis in the United States. Spinal involvement is said to occur in less than 1%. METHODS: A review of the clinic records of all patients having a diagnosis of spinal tuberculosis at the New Jersey Medical School National Tuberculosis Center from January 1994 to January 1999 was done. RESULTS: Twelve patients with spinal TB were identified (2.2%). Spinal tuberculosis is characteristically chronic and slowly progressive. Presenting symptoms are usually back and neck pain or pain at the site of involvement. Pain was noted in a majority (83%) of our patients. Despite proven diagnoses by culture or histology, only 41% were purified protein derivative-positive. Unlike pulmonary tuberculosis, which may present with a constellation of symptoms, including cough, weight loss, fever, and night sweats, if tuberculosis only involves bone, pain may be the only symptom. Most patients were successfully treated with surgery and anti-TB medications. Tuberculosis remains a major public health problem. CONCLUSIONS: Before the disease can be treated, it must be recognized. Tuberculosis should be considered a diagnostic possibility when spinal osteomyelitis is seen, even in the absence of pulmonary disease or HIV infection. Back pain in a patient with tuberculosis should be evaluated. Early initiation of an appropriate treatment regimen may reduce the need for surgical intervention, but most patients require surgery. PMID- 16357693 TI - Pediatric rheumatology for the adult rheumatologist I: therapy and dosing for pediatric rheumatic disorders. AB - Treatment of children with rheumatic diseases has advanced with novel therapeutics and the use of early aggressive treatment to achieve better long term functional outcome. Many of the current treatments in pediatric rheumatology are based on studies in adults on medications without U.S. Food and Drug Administration labeling for pediatric use. This is not ideal because the pharmacokinetics (absorption, distribution, metabolism, and elimination) of many medications when used in children vary according to age, somatic growth, sexual maturity, and ontogeny of drug-metabolizing enzymes. Special dosing, administration considerations, and toxicity screening are reviewed for the more commonly used medications. Vaccinations for children on antirheumatic drugs are also discussed. Continued study of the long-term impact of medications and biologic treatments on children is necessary, but given the paucity of children with rheumatic diseases, this will require multicentered trials and collaborations. Lastly, this article reviews recent regulatory and legislative action on pediatric drug testing. Passage of the Pediatric Research Equity Act of 2003, which requires testing of pharmaceuticals in children, will facilitate more rational use of drugs in pediatric rheumatic diseases in the future. PMID- 16357694 TI - Basic science for the clinician 34: Interleukins of current clinical relevance (part II). AB - As you saw in the first part of this description of interleukins, normal orchestration of wound healing, the protective immune response and inflammation involves many cells that must effectively communicate with each other. The means of this communication is often soluble messengers (cytokine) and many of them bear the title interleukin. Although all these messengers have a role in normal immune homeostasis, it is apparent that many are involved in tissue damage in a variety of disease, eg, rheumatoid arthritis, osteoarthritis. I dealt with interleukins (IL) 3 to 16 in the first part of this project. We now pick up the story with IL-17. Turns out, much of the most exciting recent work in rheumatology has focused on IL-17 and IL-18. The disparate effects of the interleukins may be confusing, often a single cytokine producing multiple effects seemingly at crossed purposes, but we are in our infancy when it comes to insights into the molecular biology of normal immune function, homeostasis, inflammation, and disease. PMID- 16357695 TI - The lung in systemic sclerosis. PMID- 16357696 TI - Drug-induced lupus after treatment with infliximab in rheumatoid arthritis. AB - We report a case of a 45-year-old man with an 8-month history of rheumatoid arthritis, who was treated with hydroxychloroquine 400 mg per day and 15 mg intramuscular methotrexate per week without reaching a good control of the disease. The patient was successfully treated with 3 mg/kg infliximab for 20 weeks. Before the last infusion, drug-induced lupus (DIL) was diagnosed based on the clinical features of fever > 37.5 degrees C, recurrence of active synovitis, myalgia, erythematosus rash, pericardial and pleural effusion, and of some laboratory findings (antinuclear antibodies 1:160 and anti double-strand DNA positive by DNA recombinant plasmid assay dsDNA). After infliximab discontinuation and the beginning of therapy with methylprednisolone, lupus symptoms resolved within 6 weeks. A new rheumatoid arthritis flare, occurring after 8 weeks, was controlled by methotrexate plus leflunomide. We also review the development of antinuclear and antidouble-strand DNA antibodies and drug induced lupus in patients treated with anti-TNFalpha agents (infliximab, etanercept, and adalimumab). PMID- 16357697 TI - Severe proximal myopathy and mononeuritis multiplex in rheumatoid arthritis: manifestations of rheumatoid vasculitis. AB - Vascular injury is considered to be a key finding in the pathogenesis of rheumatoid arthritis (RA). Manifestations are varied depending on the vessel size and the organ system involved. Vasculitis leading to symptomatic inflammatory myositis is a rare complication of RA. We describe a 62-year-old man with seropositive erosive RA of 1-year duration, who presented with severe proximal weakness and mononeuritis multiplex. His joint disease was clinically mild at the time of presentation. Creatine kinase was normal and the electromyogram did not suggest myopathy. However, muscle biopsy revealed extensive small vessel vasculitis and severe inflammatory myositis. This report emphasizes the importance of fully evaluating patients with RA who present with proximal myopathy. The myopathy in our patient was not related to active joint disease, disuse atrophy, or complication of therapy. Rheumatoid vasculitis leading to myositis is a rare and not well-recognized complication of RA for which aggressive immunosuppressive therapy is warranted. PMID- 16357698 TI - Recurrent pulmonary embolism despite inferior vena cava filter placement in patients with the antiphospholipid syndrome. AB - In patients at risk for pulmonary emboli, consideration is often given to placement of an inferior vena cava (IVC) filter to prevent propagation of a distal thrombus. However, long-term benefits remain controversial, and deep venous thrombosis and IVC thrombosis may result from the procedure itself. Whether a filter if beneficial or even detrimental in patients with the antiphospholipid syndrome (APS) is unclear. We reviewed clinical outcomes in 2 patients who had IVC filter placement years before the diagnosis of the APS and 1 who had a contraindication to anticoagulation. Recurrent pulmonary emboli were seen despite the presence of the filter. IVC pathology sometimes revealed thrombus both proximal and distal to the IVC filter. Pulmonary emboli in the APS may be secondary to deep venous thrombosis (DVT). They may also occur secondary to a cardiac source or in situ thrombosis in the pulmonary vessels. An IVC filter will not be of benefit if the heart or the lungs are the primary source for the emboli. It may also not protect against propagation of a more distal thrombus if collateral vessels develop around the filter or a thrombus is present on the proximal side of the filter. Recurrence of pulmonary emboli after a filter placement should alert the clinician to the possibility of a hypercoagulable state such as APS. Clinicians need to assess risks and benefits carefully before placing a permanent IVC filter in patients with APS. Whether a temporary or retrievable filter is safer in APS and more effective is unknown at the present time. PMID- 16357699 TI - Polymyalgia rheumatica as an unusual cause of pleural and pericardial effusion. AB - We report a patient with polymyalgia rheumatica (PMR) who initially presented with chest pain pleural and pericardial effusions. A history was then obtained of shoulder girdle aching and stiffness, suggestive of PMR, and laboratory investigations supported the diagnosis. A rapid response to 15 mg prednisolone once daily was noted, with resolution of effusion and symptoms. Although pericardial effusion has previously been reported, this is the first reported case of pleuropericardial effusions in PMR. Rheumatologists should be aware that PMR may occasionally be the explanation for pleuropericardial effusion in the elderly. PMID- 16357700 TI - Whiplash: a case of regretting being responsible for naming a condition. PMID- 16357702 TI - Associated autoimmune disorders in patients with classic lupus erythematosus. PMID- 16357703 TI - Nail fold capillaroscopy evaluation pearl. PMID- 16357704 TI - Osteoporosis treatment after hip fracture: slow progress. PMID- 16357705 TI - Secondary prevention of hip fractures among the hospitalized elderly: are we doing enough? AB - BACKGROUND: Older individuals with hip fractures almost always have osteoporosis. Such individuals are at increased risk of experiencing other osteoporotic fractures, including recurrent hip fractures. The management of such patients should include assessing bone mineral density and treating osteoporosis. OBJECTIVE: The objective of this study was to investigate if elderly (> or =65 years) patients with hip fractures were assessed and treated for osteoporosis. METHODS: A retrospective chart review was conducted of all elderly patients who underwent hip fracture surgery at a university teaching hospital during the calendar years 1997 to 1999. RESULTS: A total of 95 subjects were identified (29% males and 71% females). Subjects' age ranged from 65 to 96 years with a mean (+/ standard deviation) 81 +/- 7 years. Femoral neck fractures were the most common (51%), followed by intertrochanteric (43%) and subtrochanteric fractures (3%). Two subjects (2%) had fractures at multiple sites. The most common cause of a hip fracture was a fall (87%). Other causes included motor vehicle accidents (6%) as well as other trauma (4%). One subject had a spontaneous hip fracture. A history of hip fractures was obtained in 8% of subjects. Osteoporosis was diagnosed in 17% of subjects before admission. On admission, 9% of subjects were receiving calcium, 3% were receiving vitamin D, none were receiving alendronate, and 1 subject was receiving calcitonin. Approximately 3% of female subjects were receiving estrogen on admission. On discharge, 11% of subjects were prescribed calcium, 6% were prescribed vitamin D, none were prescribed alendronate, and 2% were on calcitonin. None of the female subjects were discharged on estrogen. During hospitalization, 88% of subjects who were admitted to nonmedical services were seen by either a geriatric or a general internal medicine consult. Obtaining a medical and/or geriatric consult did not have an apparent effect on the frequency of treating osteoporosis in this high-risk group of subjects. CONCLUSION: Older adults with hip fractures are not adequately treated for osteoporosis. This places them at increased risk of other osteoporotic fractures, including recurrent hip fractures. PMID- 16357706 TI - Pachydermodactyly: six new cases from China. AB - BACKGROUND: Pachydermodactyly is an infrequently reported disease characterized clinically by painless swelling of the soft tissues at interphalangeal joints. Histopathologic features include increased dermal accumulation of collagen fibers. Young males are predominantly affected. There are only approximately 50 cumulative cases reported worldwide. OBJECTIVE: : This is a review of 6 new cases of pachydermodactyly from our center. METHODS: Characteristics of our cases are described in detail and the literature of this disease is reviewed. RESULTS: The mean disease onset age of our 6 patients was 14 +/- 2.7 years; 5 of the patients were male and 1 was female. Clinical presentations included chronic noninflammatory bulbous swelling of proximal interphalangeal (PIP) joints. Mean numbers of involved joints were 5 +/- 3; most frequently affected were PIP2, 3, and 4 symmetrically. There was 1 patient with monoarticular involvement. X-ray demonstrated soft tissue swelling with no bony or articular damage. Histopathology displayed increased dermal deposition of collagen fibers. CONCLUSIONS: Pachydermodactyly is not well recognized by rheumatologists and should be differentiated from inflammatory arthritis involving PIP joints. PMID- 16357707 TI - Adult-onset Still disease in southeast Brazil. AB - BACKGROUND: Adult-onset Still disease (AOSD) has been described all over the world. Clinical presentations and prognosis have varied in different studies. OBJECTIVE: The objective of this study was to determine the clinical presentation and the evolution of AOSD at a tertiary referral center in southeast Brazil. METHODS: The clinical records of 16 patients were retrospectively studied to determine symptoms at diagnosis, follow up, and the medication prescribed. RESULTS: The mean age at onset was 30.8 years (range, 24-55 years; standard deviation [SD], 9.2 years) with a slight male prevalence (54.2%). All patients presented constitutional symptoms, fever, and skin rash. Liver involvement was observed in all cases, with hepatomegaly in 81.3%, increased liver enzymes in 50.0%, and hypergammaglobulinemia in 68.8%. Cardiac involvement was observed in 12.6%, pleuritis in 6.3%, and renal involvement in 25.0%. All patients presented leukocytosis with a predominance of neutrophils. Elevated ferritin levels were observed in 56.3%, and these levels were normalized after disease remission. Initial treatments included nonsteroidal antiinflammatory drugs and low-dosage corticosteroids in all patients; 43.8% also needed methotrexate. In 25.0% of cases, a monocyclic disease was observed; others had recurrent episodes. After a follow up of 6.9 years (SD, 1.2 years), carpal ankylosis was the main articular sequel, observed in 53.6% of the patients. CONCLUSION: AOSD is rare in southeast Brazil. Although less severe systemic manifestations, like serositis and pneumonitis, were observed, reversible liver involvement was common; the frequency of recurrent disease and carpal ankylosis was higher than in previous studies. PMID- 16357708 TI - Audiovestibular disorders in patients with ankylosing spondylitis. AB - OBJECTIVE: Audiovestibular disorders have been described in several autoimmune diseases but have not been studied in patients with ankylosing spondylitis (AS). The aim of this study was to evaluate the audiovestibular function in patients with AS. METHODS: We prospectively evaluated 22 consecutive patients with AS. Clinical, radiologic, and immunogenetic features were analyzed. All patients underwent a complete ear, nose, and throat physical examination and audiologic evaluation that included pure-tone audiometry thresholds at octave frequencies of 250 to 8000 Hz, impedance audiometry (tympanogram, static compliance, acoustic reflexes, and reflex decay), and speech audiometry. Thirty-one healthy volunteers and 26 patients with rheumatoid arthritis (RA) were included as controls. RESULTS: Patients with AS had a median age of 45.5 years (interquartile range [IQR] 38-54) and a median disease duration of 20 years (IQR 12.5-26.2). Otosclerosis was observed in 2 patients with AS (9.1%), 3 patients with RA (11.5%), but not in any healthy controls (P = not significant). Sensorineural hearing loss (SNHL) was observed in 13 patients with AS (59%), 17 patients with RA (65.4%), and 13 healthy controls (41.9%) without statistically significant differences. Because age can influence audiometric results, patients and controls were divided into 4 age groups. A higher frequency of SNHL was observed in patients with AS from 45 to 59 years compared with healthy controls and patients with RA (87% versus 20%, P = 0.03, versus 70%, P = not significant, respectively). CONCLUSION: Middle-aged patients with AS had a significantly higher frequency of SNHL compared with controls. SNHL should be considered as a possible complication of AS as well as of RA and other autoimmune diseases. Possible toxic effect of nonsteroidal antiinflammatory drugs should also be considered in such patients. PMID- 16357709 TI - Coexisting systemic lupus erythematosus and sickle cell disease: a diagnostic and therapeutic challenge. AB - Systemic lupus erythematosus (SLE) and sickle cell disease (SCD) are relatively common disorders with comparable prevalence among blacks. The coexistence of these 2 conditions in the same individual appears to be rare. We report 4 cases of coexisting SLE and SCD. These patients displayed a broad spectrum of musculoskeletal, central nervous system, and renal complications that may be associated with either SCD or SLE. Because of a substantial overlap between the clinical manifestations of these 2 disorders, the diagnosis of SLE in patients with SCD may be difficult to establish and is often delayed. Up to 23% of patients with SCD may have antinuclear antibodies. All patients in this series had antecedent SCD but new important complications from SLE. PMID- 16357710 TI - Critical care myopathy in a child. AB - Critical care myopathy (CCM) is an important, often unrecognized cause of weakness in the intensive care unit (ICU). We report a child who developed severe myopathy in the ICU. Our patient was newly diagnosed to have autoimmune hepatitis that may have predisposed her to toxicity from corticosteroids and neuromuscular blocking agents, but she recovered her strength completely. Risk factors that may have predisposed this patient to CCM include underlying liver dysfunction, presence of acute lung injury, use of corticosteroids, and use of neuromuscular blockade. Given the apparent prevalence and underreporting of CCM in the ICU, it is important for rheumatology consultants to include this entity in their differential diagnosis of weakness in critically ill children and adults, and to recommend appropriate evaluation. PMID- 16357711 TI - Erythema elevatum diutinum in the setting of connective tissue disease and chronic bacterial infection. AB - Erythema elevatum diutinum (EED) is a rare and chronic cutaneous leukocytoclastic vasculitis. It is predominantly seen on the extensor surfaces of the extremities. Although a specific cause is largely unknown, EED has been noted to occur in association with a wide variety of diseases. A 28-year-old man with systemic lupus erythematosus (SLE) and a 53-year-old woman with an overlap syndrome of rheumatoid arthritis and polymyositis are presented. Both patients developed EED in the setting of chronic recurrent bacterial infections. Patients with a connective tissue disease are at increased risk for such infections secondary to immunosuppression, either from the disease itself or secondary to immunosuppressive therapy. EED has been independently reported to occur in the setting of connective tissue disease as well as in the setting of chronic infection. Our patients had both of these underlying conditions, which are known to predispose patients to immune complex-mediated vasculitides, in this case EED. One patient's EED responded to treatment of the SLE and the other improved, as has been previously reported with dapsone. PMID- 16357713 TI - Bilateral shortening of third and fourth metacarpal bones. PMID- 16357712 TI - Vertebral sarcoidosis mimicking lytic osseous metastases: development 16 years after apparent resolution of thoracic sarcoidosis. AB - Vertebral sarcoidosis is a rare condition that can present with persistent back pain, often with concurrent lung, lymph node, or skin involvement. It can produce lytic or blastic osseous lesions that are indistinguishable from metastatic cancer on bone scan and magnetic resonance imaging (MRI). It usually occurs at the time of initial diagnosis of sarcoidosis, but may in very rare cases appear many years after presumed resolution of thoracic sarcoidosis. We present the case of a 47-year-old man who developed persistent low back pain 16 years after spontaneous resolution of stage I pulmonary sarcoidosis. MRI of the spine showed lytic thoracic and lumbar vertebral lesions. Computed tomography of the chest showed a pleural-based lung mass, multiple pulmonary nodules, and hilar and mediastinal lymphadenopathy. Positron emission tomography with fluorodeoxyglucose was widely positive, including at the vertebral foci noted on MRI. Metastatic lymphoma was suspected, but mediastinal lymph node and vertebral body biopsies showed noncaseating granulomas with negative stains for acid-fast bacilli and fungi. After 1 month of treatment with prednisone, the angiotensin-converting enzyme level and erythrocyte sedimentation rate had normalized, and the back pain was substantially improved. We found only 1 case report of a longer interval between resolution of initial sarcoidosis and development of vertebral involvement. PMID- 16357714 TI - Septic joint successfully treated with arthrotomy: 1861. PMID- 16357715 TI - Basic science for the clinician 26: Tolerance--mechanisms and manifestations. AB - Usually the immune system monitors our internal milieu, protecting us from external and internal risks, silently watching over the rest. Pathologic autoimmunity is usually avoided, except in certain animal models and certain immunogenetically predisposed people. On the other hand, "salutary autoimmunity" occurs in the interaction of immune systems with themselves or other immune pathways as part of control or recognition mechanisms, eg, idiotype network, epitope-MHC complex binding with B- or T-cell antigen receptors. There are means by which dangerous autoimmune reactants are eliminated from the repertoire before birth; some of these are described here. However, potentially dangerous autoimmune effectors can be identified in the immunologic repertoire of certain adult animal strains but not damage the animal; these latent autoimmune effector pathways are held in check in adult animals/humans by active suppressive mechanisms.The identification and control of autoimmunity may well be the holy grail of rheumatology. Autoimmunity may also be an active participant in a number of other organ systems and diseases, so lessons learned in immunology may apply broadly throughout medicine. Once a better understanding of the normal processes and aberrancies that lead to disease are had, one can look forward to interventions to reestablish tolerance, perhaps by something as simple as a capsule containing the appropriate self antigen-containing molecules--"oral tolerizing" as a cure for autoimmune diseases! There is much promising evidence in favor of the efficacy of oral tolerance in animal models, but as of yet, human disease has proven a harder nut to crack. Although the concept of oral tolerance is nearing its centennial, there is still much to learn! PMID- 16357717 TI - Pneumonitis caused by Legionella pneumoniae in a patient with rheumatoid arthritis treated with anti-TNF-alpha therapy (infliximab). PMID- 16357718 TI - Listeria monocytogenes septic arthritis in a patient treated with etanercept for rheumatoid arthritis. PMID- 16357719 TI - Primary esophageal melanoma in a patient with limited scleroderma. PMID- 16357720 TI - A case of dermatomyositis that developed after delivery. PMID- 16357722 TI - Foreword: dialogues in rheumatology. PMID- 16357723 TI - Section 3: The nature of pain: pathophysiology. AB - An understanding of the pathophysiology of pain involves the concepts of neuronal plasticity at the levels of the nociceptor neurons, spinal cord, and brain. Neuroplasticity allows the neurons in the brain to compensate for injury and adjust their activity in response to new situations or changes in their environment. This is an important physiologic event in the development of central sensitization. Other concepts include modulatory effects at the nociceptor, sympathetically mediated pain, the "wind-up" phenomenon, central sensitization, and descending and ascending central modulatory mechanisms for the perception of pain, as well as the related pain motivations and behaviors. Numerous modulatory mechanisms for pain have been postulated that control the degree of pain perceived and the emotional and behavioral phenomena associated with a patient's pain experience. These numerous mechanisms take place at all levels of the nervous system: peripheral nerves, spinal cord, and brain. Despite great advances in unraveling the complexities of the pathophysiology of pain, much remains to be discovered. It is hoped that further research into this critical area will lead to better therapies. PMID- 16357724 TI - Section 4: treating the patient in pain. AB - Physicians may choose from a variety of pharmacologic and nonpharmacologic options to treat patients with painful rheumatic diseases. Osteoarthritis (OA) is the most common type of arthritis requiring pain management. New disease modifying antirheumatic drugs and biologic response modifiers can improve disease states in patients with rheumatoid arthritis (RA). After the inflammatory component of RA is minimized with such agents, treatment goals shift to those similar to secondary OA and other degenerative joint diseases. Relief of pain and improvement in functional status are essential components of effective therapy. A pure analgesic such as acetaminophen and nonsteroidal antiinflammatory drugs, including the cyclooxygenase-2-selective inhibitors for those at risk for gastrointestinal side effects, may be used at the lowest effective doses. Combination therapy for acetaminophen and an opioid may maximize pain relief and provide greater speed and duration of action than the separate components. Use of the atypical opioid tramadol with acetaminophen often results in an improved side effect profile compared with stronger opioids, with similar levels of pain relief. Adjunctive therapy with agents such as topical analgesics, intraarticular hyaluron, tricyclic antidepressants, anticonvulsants, muscle relaxants, and anxiolytics may also be helpful. Nonpharmacologic therapies such as exercise, physical therapy, and psychologic counseling may also diminish pain and improve outcome in patients with rheumatic diseases. One may also consider yoga, acupuncture, biofeedback, massage, relaxation techniques, and other alternative therapies. PMID- 16357725 TI - Section I: Introduction. PMID- 16357726 TI - Section 5: Conclusions and summary. PMID- 16357728 TI - Section 2: The importance of improving function in patients with pain. AB - When evaluating pain in the rheumatic diseases, assessment of adequate therapy has expanded from purely biochemical measures and joint counts to determining the degree of function. Because pain is an important predictor of function, instruments have been developed that attempt to measure how disease and pain affect parameters such as activities of daily living, quality of life, ability to work, and emotional well-being. Some measures, such as visual analogue scales, measure only the intensity of pain; multidimensional scales assess a variety of factors related to the patient's overall ability to function. The Short Form-36 measures the impact of disease on overall quality of life; adaptations have been made to some specific diseases. The Health Assessment Questionnaire was one of the first multidimensional questionnaires developed for rheumatic diseases, and this has been further refined for rheumatoid arthritis with the Multidimensional Health Assessment Questionnaire. PMID- 16357729 TI - Reversible posterior leukoencephalopathy syndrome in systemic lupus erythematosus: what can this teach us? PMID- 16357730 TI - Correction of allopurinol dosing should be based on clearance of creatinine, but not plasma creatinine levels: another insight to allopurinol-related toxicity. AB - BACKGROUND: Dosing of allopurinol should be corrected depending on renal function, but corrections based on either plasma creatinine (Pcr) or creatinine clearance (CrCl) have been suggested to be minimal standards of care. METHODS: Data from a cohort database of 484 gouty patients were used to calculate estimated allopurinol doses using CrCl and estimation of the clearance of creatinine using the equation of Cockroft and Gault (CrCl-CG) if, as a hypothesis, a dosage of 300 mg/d would be prescribed in any patient with Pcr <2.0 mg/dL. Also, allopurinol-related toxicity previous to rheumatologic consultation, during previous allopurinol therapy, and the relationship between both and estimated allopurinol doses were reviewed. RESULTS: The cutoff point of plasma creatinine <2 showed 13% sensitivity and 100% specificity to detect CrCl <50 mL/min. Correlation and agreement between CrCl and CrCl-CG were good, as was the correlation between corrected doses using CrCl and CrCl-CG. One third of patients with Pcr 1.0-1.5 mg/dL and 90% of those with Pcr 1.5-2.0 mg/dL would receive estimated doses over 400 mg/dL/d CrCl. Also, 10% and 34% would receive estimated doses over 600 mg/dL/d CrCl, respectively. Allopurinol-related toxicity previous to consultation (11%) was associated with estimated doses over 400 mg/dL/d CrCl and severe toxicity with estimated doses over 600 mg/dL/d CrCl. When patients were given doses corrected on CrCl, few side effects were observed during follow up (6.7%), and the only severe one was associated with corrected dose over 600 mg/d. CONCLUSIONS: Dosage adjustment of allopurinol should be based on clearance of creatinine or estimation of glomerular filtration using the Cockcroft-Gault equation. Pcr is insensitive enough to detect renal function impairment so that patients may be placed at risk for overdosing side effects. Corrected doses over 600 mg/dL/d CrCl may be associated with increased risk of severe toxicity. PMID- 16357731 TI - Corneal melt in rheumatic disorders: effect of disease-modifying antirheumatic drugs on morbidity. AB - OBJECTIVE: The objective of this study was to review the characteristics of patients with corneal melt and to assess if treatment with disease-modifying antirheumatic drugs (DMARDs) improved the visual outcome. METHOD: We did a retrospective analysis of patients diagnosed with corneal melt between 1976 and 2002. Twenty-one patients with rheumatoid arthritis and 5 patients with primary Sjogren syndrome (26 patients, 42 eyes) were included in the analysis of visual outcome. Visual outcome was described as "fair" if the corrected visual acuity was 20/200 or better and as "poor" if the corrected visual acuity was worse than 20/200. RESULTS: Visual outcome was fair in 9 patients (90%) in the DMARD group versus 2 patients (13%) in the no DMARD group (P= 0.001), and in 14 eyes (93%) in the DMARD group versus 7 eyes (26%) in the no DMARD group (P = 0.001). CONCLUSION: The use of DMARDs improves visual outcome in patients with corneal melt. PMID- 16357732 TI - Prevalence of abuse in fibromyalgia and other rheumatic disorders at a specialized clinic in rheumatic diseases in Guatemala City. AB - BACKGROUND: The importance of past adverse experiences is increasingly recognized in patients with rheumatic disease. OBJECTIVE: The objective of this study was to study the association of physical, verbal, and sexual abuse in patients with rheumatic disorders as compared with healthy volunteers. METHODS: In this case control study, 500 new patients attending an outpatient rheumatic clinic were interviewed from September 1, 1999, to August 31, 2001. A total of 187 patients with 3 diagnoses were selected: 58 had fibromyalgia (FM), 74 rheumatoid arthritis (RA), and 55 patients with soft tissue rheumatic disease (STRD). All selected patients were asked to complete a questionnaire designed to obtain information regarding demographics and history of verbal, physical, and sexual abuse. A group of 187 healthy control subjects were also included, matched for sex and age. RESULTS: The prevalence of abuse was significantly more common in the rheumatic disease group than in the control group (48.1% versus 15%, P < 0.001). The prevalence of abuse among the groups was as follows: 70.7% of patients with FM reported abuse (24.3% verbal, 60.9% physical, and 14.8% sexual), 35.1% of patients with RA had a history of abuse (42.3% verbal, 30.7% physical, and 0% sexual), whereas 41.8% of patients with STRD reported abuse (43.4% verbal, 43.4% physical, and 0% sexual). When comparing the 3 groups, patients with FM showed a higher prevalence of abuse (P < 0.05). The abuse was usually longstanding (range, 1-10 years), and most abusers were close family members. CONCLUSION: Abuse, both physical and psychologic, was significantly increased in our rheumatic disease population, especially in patients with FM. Further studies are needed to fully establish its role. Questions about abuse may provide important information relative to care of our patients. PMID- 16357734 TI - Antibodies to cyclic citrullinated peptide in patients with chronic arthritis attending an arthritis-monitoring clinic. AB - BACKGROUND: Early diagnosis of rheumatoid arthritis allows prompt initiation of antirheumatic therapy, which is associated with improved outcome. The use of IgM rheumatoid factor, the most widely used serologic test in assisting the diagnosis of rheumatoid arthritis, is limited by low specificity. An enzyme-linked immunosorbent assay test detecting antibodies to cyclic citrullinated peptide (CCP) has been developed with apparently higher specificity for rheumatoid arthritis. AIM: We aimed to evaluate an assay for anti-CCP antibodies by determining the prevalence and titer of antibodies to CCP in a group of patients with chronic arthritis. METHOD: Thirty-four sera were collected from outpatients attending an arthritis-monitoring clinic and tested for anti-CCP and IgM rheumatoid factor. RESULTS: Anti-CCP and rheumatoid factor were detected in 86% and 72% of patients with rheumatoid arthritis, respectively. Six patients negative for rheumatoid factor were positive for anti-CCP. CONCLUSION: Anti-CCP antibodies are frequently detectable in high titer in patients with longstanding rheumatoid arthritis. This assay may be especially helpful in such cases when positive in rheumatoid factor-negative patients. PMID- 16357733 TI - Adult-onset acute rheumatic fever: possible resurgence in southern Taiwan. AB - BACKGROUND: By the late 1980s, acute rheumatic fever (ARF) had become a rare disease in Taiwan. The low prevalence rate in this area is attributed to a better economic status, which has led to improved public health and adequate medical services. OBJECTIVES: The increasing number of patients with adult-onset ARF in the United States described in the literature prompted us to evaluate the cases diagnosed in our medical center. METHODS: A retrospective chart review was performed for patients with arthritis from July 1988 to October 2004. To be included, patients had to meet revised Jones criteria. RESULTS: Three adult patients with ARF have been diagnosed since June 2001, with no childhood ARF being diagnosed. All cases presented with migratory polyarthritis, whereas 1 had erythema marginatum and transient carditis. These patients responded well to treatment with antibiotics and nonsteroidal antiinflammatory drugs. CONCLUSIONS: Clinicians must provide careful assessment and treatment to patients presenting with acute pharyngitis. A possible resurgence of ARF can be eradicated by primary prevention of streptococcal pharyngitis. PMID- 16357735 TI - Retrospective review of the clinical manifestations and outcomes in Puerto Ricans with idiopathic inflammatory myopathies. AB - BACKGROUND: The idiopathic inflammatory myopathies (IIM) are a heterogeneous group of conditions characterized by chronic inflammation of muscles, resulting in skeletal muscle weakness. Racial differences are apparent in the clinical manifestations and outcome of IIM. No previous studies have been conducted to evaluate the clinical manifestations and outcome of Puerto Ricans with IIM. OBJECTIVE: The objective of this study was to describe demographic characteristics, clinical features, and functional outcome of Puerto Rican patients with IIM. METHODS: Medical records at a university medical center were reviewed retrospectively to collect data regarding demographic factors, initial presentation, diagnostic tests, serologic markers, treatment, and disease course. A Functional Grading Scale was administered to 40 of the patients with IIM to determine their functional status. RESULTS: Fifty patients with IIM were identified. Eight patients had polymyositis (PM), 23 patients had dermatomyositis (DM), 2 patients had DM/PM with coexistent malignancy, 12 patients had juvenile DM, and 5 patients had DM/PM with an associated other rheumatologic disease. At diagnosis, proximal muscle weakness was present in all patients, 15 (30%) had myalgias, 5 (10%) patients had dysphagia, and 87% had elevation of serum muscle enzymes. Visceral involvement was rare. Interstitial fibrosis was identified in only 2 patients (1 juvenile DM and 1 PM). Twenty-six patients (51%) achieved complete remission. The mean score for the Functional Grading Scale was 28.6 (maximum 30). The lowest scores were seen for patients with disease duration of less than 2 years. CONCLUSION: Puerto Ricans with IIM in this survey showed a low presence of visceral involvement, high remission rate, and low mortality. PMID- 16357736 TI - Fibromyalgia frequency in hepatitis B carriers. AB - BACKGROUND: Fibromyalgia (FM) is characterized by diffuse musculoskeletal pain, fatigue, morning stiffness, and sleep disturbance. Chronic viral infections may trigger FM symptoms. OBJECTIVES: In this study, we aimed to evaluate whether there was an association between HBsAg seropositivity and fibromyalgia syndrome. METHODS: Fifty hepatitis B carriers (HBsAg positivity and anti-HBs negativity in sera for at least 6 months) and 50 age- and sex-matched HbsAg-negative control subjects were enrolled in this study. The hepatitis B carriers with normal or slightly elevated aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were recruited from the infectious diseases outpatient clinic and the control group was recruited from the physical medicine and rehabilitation outpatient clinic. The relationship between groups was calculated by independent Student t test, chi-squared test, and Fisher exact test for comparing proportions. Alpha criterion for significance was set at P < 0.05. RESULTS: There was no statistically significant difference between the groups according to sex, mean age, body mass index, serum ALT, and AST levels (P > 0.05). FM syndrome and FM-associated symptoms were much more prevalent in the hepatitis B group (P < 0.001). CONCLUSION: The present study suggests that chronic hepatitis B carriage appears to increase the risk of FM and many of the typically associated symptoms. Whether this association is related to altered liver function, viral infection, concerns associated with chronic disease, or other factors, physicians should be aware of this apparent association. PMID- 16357737 TI - Nontropical pyomyositis in an immunocompetent host. AB - Pyomyositis is a primary infection of the striated muscle. We describe the clinical and imaging features of 2 cases with nontropical pyomyositis in immunocompetent hosts. Staphylococcus aureus was the causative agent; the treatment included antibiotics and either computed tomography or magnetic resonance-guided percutaneous needle drainage, avoiding open drainage. This treatment modality was successful with complete recovery of movement. Because of the rarity of pyomyositis in temperate climates, the common lack of specific signs or symptoms, and the frequent negative blood cultures, considerable delay precedes the diagnosis. Fever, elevated erythrocyte sedimentation rate, and muscle stiffness are diagnostic clues. Increased awareness, especially in immunocompetent hosts, should lead to earlier diagnosis and treatment with improved outcomes. Diagnosis and treatment can be reached at the same time in some cases by image-guided percutaneous drainage. PMID- 16357738 TI - Reversible posterior leukoencephalopathy syndrome in systemic lupus erythematosus with thrombocytopenia treated with cyclosporine. AB - Thrombocytopenia is one of the common manifestations of systemic lupus erythematosus (SLE). Its treatment consists of corticosteroids and/or other immunosuppressive drug such as azathioprine, vincristine, cyclosporine, and mycophenolate mofetil. Common adverse reactions of immunosuppressive treatment are infections, hematologic and liver function abnormalities, but neurologic complications may also be seen. Reversible posterior leukoencephalopathy syndrome (RPLS) is a syndrome manifested by headache, nausea, vomiting, seizures, cortical blindness, and visual disturbances. Neuroimaging shows bilateral subcortical and cortical edema with prominent posterior distribution. Moreover, treatment with immunosuppressive drugs such as cyclosporine, cisplatin, tacrolimus, and interferon-alpha can induce a condition resembling RPLS. We report a case of a young woman with SLE and thrombocytopenia, who developed severe perspiration, headache, and seizure after receiving cyclosporine. A brain magnetic resonance image showed multiple high signal intensities on T2-weighted images predominantly located at the parietooccipital lobe. The patient recovered completely clinically and radiologically after discontinuing cyclosporine. PMID- 16357739 TI - Ulcerative necrosis of the glans penis resulting from polyarteritis nodosa. AB - Polyarteritis nodosa (PAN) may be systemic or isolated in distribution and may involve virtually any organ or tissue in the body. We report an unusual case of PAN with the clinical features of both penile and renal involvement in a 21-year old man diagnosed incidentally by pathologic examination of the nephrectomized left kidney. Systemic and surgical therapeutic procedures were applied and his symptoms resolved within 6 weeks. Penile ulceration is a rare but serious manifestation of PAN that needs prompt and aggressive treatment. PMID- 16357740 TI - Diagnosis of Takayasu arteritis. PMID- 16357741 TI - Chondrocalcinosis in a 180-year-old cadaver. PMID- 16357743 TI - Basic science for the clinician 27: Toll-like receptors and nucleotide oligomerization domains. AB - Ancient protective mechanisms are in place, deep within our defenses against infection and malignancy, often unappreciated until homologous proteins found within less phylogenetically advanced organisms are identified. Such is the case with 2 major recent finds, the Toll-like receptors (TLRs) and nucleotide oligomerization domain (NOD) families of innate immunity molecules. These families of receptors have high specificity, limited heterogeneity, and no plasticity; nonetheless, they play a pivotal role in rapid initial defenses against pathogens. Moreover, studies of the mechanisms of TLRs and NODs show how they and IL-1 and IL-18 stand at the threshold of the adaptive immune response and help to accelerate specific immune responsivity. Nonspecific reactivity of these preprogrammed receptors may be how relatively nonpathogenic organisms like yersinia and chlamydia may drive the inflammation of reactive arthritis and atherosclerosis. The inflammation of rheumatoid arthritis may be magnified, if not initiated, by these innate mechanisms as well. PMID- 16357744 TI - Anti-tumor necrosis factor therapy is tolerated in an individual with homozygous complement C2 deficiency. PMID- 16357745 TI - Hip arthroplasty in a 101-year-old patient. PMID- 16357746 TI - Coxopathy in congenital afibrinogenemia. PMID- 16357748 TI - Introduction. PMID- 16357749 TI - Immunologic mechanisms in the pathogenesis of rheumatoid arthritis. AB - Although much is known about the etiology and pathogenesis of rheumatoid arthritis (RA), our understanding of the immune pathways remains incomplete. The observed clinical and pathologic manifestations result from activation of several interrelated immune pathways. Current concepts of RA pathogenesis, supported by animal models, laboratory studies, and clinical observation, have reestablished and revised some of the original views. Early proposals emphasized the importance of autoantibodies and immune complexes in the initiation of RA, suggested a role for T cells in the inflammatory response characteristic of RA, and based disease perpetuation on an imbalance in the cytokine networks. We now recognize that each of these interrelated mechanisms significantly contributes to RA pathogenesis, including T cells that can help initiate and perpetuate the disease. This article reviews the major components and immune pathways involved in RA and briefly discusses the animal models that contribute to our understanding. Although a unified theory of RA pathogenesis may not be possible at this time, a paradigm is presented that considers the immune pathways that contribute to disease progression and joint destruction. These pathways may have important implications for treatment, because their modulation by biologic response modifiers (BRMs) directed toward specific targets provides benefits to patients with RA. BRMs are a new class of therapeutic agents derived from biologically active molecules and designed to modulate specific immune or inflammatory pathways. Although currently approved BRMs still have limitations, choosing an appropriate target, possibly early rather than late in the immune response, might result in new and improved therapies for RA. PMID- 16357750 TI - Biologic therapies in clinical development for the treatment of rheumatoid arthritis. AB - The therapeutic objective in patients with rheumatoid arthritis (RA) is reduction of disease activity with an ultimate goal of disease remission. Limitations of currently available disease-modifying antirheumatic drugs and biologic therapies suggest that there remains an unmet need for agents that advance these goals in a greater proportion of patients. Progress in our understanding of the regulatory molecules and pathways that mediate the immune and inflammatory responses necessary for the initiation and perpetuation of RA has led to the identification of new targets for therapy. It is expected that the therapeutic modulation of these targets, which include proinflammatory cytokines, T and B cells, adhesion molecules, chemokines, and intra- and extracellular signaling pathways, can provide new treatment strategies in patients with RA and other autoimmune disorders. Toward this end, a series of novel agents with diverse mechanisms of action are in development. Although many of these agents are still beyond the clinical horizon, several of them have shown promise in recent trials. This article reviews a few of the many treatment strategies currently being evaluated, which are hoped to lead to greater benefits and better disease management in the clinical setting. PMID- 16357751 TI - Selective costimulation modulators: a novel approach for the treatment of rheumatoid arthritis. AB - T cells have a central role in the orchestration of the immune pathways that contribute to the inflammation and joint destruction characteristic of rheumatoid arthritis (RA). The requirement for a dual signal for T-cell activation and the construction of a fusion protein that prevents engagement of the costimulatory molecules required for this activation has led to a new approach to RA therapy. This approach is mechanistically distinct from other currently used therapies; it targets events early rather than late in the immune cascade, and it results in immunomodulation rather than complete immunosuppression. The fusion protein abatacept is a selective costimulation modulator that avidly binds to the CD80/CD86 ligands on an antigen-presenting cell, resulting in the inability of these ligands to engage the CD28 receptor on the T cell. Abatacept dose dependently reduces T-cell proliferation, serum concentrations of acute-phase reactants, and other markers of inflammation, including the production of rheumatoid factor by B cells. Recent studies have provided consistent evidence that treatment with abatacept results in a rapid onset of efficacy that is maintained over the course of treatment in patients with inadequate response to methotrexate and anti-tumor necrosis factor therapies. This efficacy includes patient-centered outcomes and radiographic measurement of disease progression. Abatacept has also demonstrated a very favorable safety profile to date. This article reviews the rationale for this therapeutic approach and highlights some of the recent studies that demonstrate the benefits obtained by using abatacept. This clinical experience indicates that abatacept is a significant addition to the therapeutic armamentarium for the management of patients with RA. PMID- 16357754 TI - Development of a Spanish-language version of the Shoulder Disability Questionnaire. AB - BACKGROUND: The Shoulder Disability Questionnaire (SDQ) is a validated shoulder functional scale that has not yet been translated into Spanish. OBJECTIVE: The objective of this study was to develop and test the consistency of a Spanish translation of The Netherlands version of the SDQ (NSDQ). METHODS: A backtranslation process was used to generate a Spanish version of the NSDQ, called the S-SDQ. Subsequently, both the NSDQ and S-SDQ were administered to 30 bilingual (English/Spanish) subjects. Finally, the S-SDQ was administered to 35 monolingual Spanish-speaking subjects with shoulder pain on 2 occasions. External consistency between NSDQ and S-SDQ scores in the bilingual subjects, and between scores for the 2 S-SDQ applications in the monolingual subjects was assessed using the intraclass correlation coefficient (Ri). Internal consistency of the S SDQ was assessed with Cronbach's alpha coefficient. RESULTS: In the bilingual subjects, the Ri between the NSDQ and S-SDQ scores was 0.991 (95% confidence interval [CI], 0.981-0.995). In the monolingual Spanish-speaking patients with shoulder pain, the Ri between the 2 S-SDQ scores was 0.99 (95% CI, 0.997-0.999). The Cronbach's alpha coefficient for the S-SDQ was 0.99. CONCLUSIONS: A highly consistent Spanish version of the NSDQ, the S-SDQ, was developed. This can be confidently used in clinical care and research settings for measurement of the functional status of shoulder joint disorders in Spanish-speaking subjects. PMID- 16357755 TI - Analysis of a cluster of cases of Wegener granulomatosis. AB - BACKGROUND: Wegener granulomatosis is a chronic inflammatory autoimmune disease of unknown etiology. The sporadic occurrence, lack of familial or genetic associations, and rising incidence suggest possible exposure to environmental agents as causative for this disease. OBJECTIVE: The objective of this study was to examine possible environmental triggers of Wegener granulomatosis. METHODS: While conducting an environmental survey of potential precipitants of Wegener granulomatosis on a cohort of patients seen at Doylestown Hospital and at the University of Pennsylvania, we identified a cluster of cases in the Dublin, Pennsylvania, region. Through hospital records and patient contacts, we located 7 cases diagnosed in a 3-year period within a 10-mile radius of an Environmental Protection Agency (EPA) Superfund toxic waste site. The radius of inclusion represents a population of approximately 50,000 individuals. Assuming complete ascertainment of cases--which is unlikely given the methods used to acquire patients--the prevalence is 2- to 4-fold greater than the expected rate of 3 per 100,000. We identified toxins at or above "action level" within the demarcated geographic region using published data from the EPA. Furthermore, we queried patients regarding their particular chemical exposures. RESULTS: These patients with Wegener granulomatosis were possibly exposed to high levels of trichloroethylene (TCE), vinyl chloride, methyl tertiary-butyl ether (MTBE), dichloroethene (DCE), and chromic acid from several industrial waste sites within the area. Additionally, these patients reported a total of greater than 30 possible exposures, including the aforesaid chemical contaminants. Three of 5 patients whose water source is known had well water that exposed them to industrial runoff and necessitated EPA intervention. CONCLUSION: This data, along with other epidemiologic studies, suggest possible toxic exposures as potentially correctable risk factors for Wegener granulomatosis. We encourage clinicians to seek data that suggests an unusual environmental exposure and to solicit information that might implicate an industrial source for these exposures. PMID- 16357757 TI - Resolution of hypertensive encephalopathy and scleroderma renal crisis with an angiotensin receptor blocker. AB - The use of an angiotensin II type-1 receptor blocker for scleroderma renal crisis is controversial. We describe a 46-year-old woman presenting with a seizure secondary to hypertensive encephalopathy as the initial manifestation of scleroderma renal crisis. She had complete resolution of end organ damage with use of an angiotensin II type-1 receptor blocker. There may be a role for angiotensin II type-1 receptor blockers in the setting of scleroderma renal crisis with central nervous system involvement. PMID- 16357756 TI - Gastrointestinal symptoms and their association with health-related quality of life of children with juvenile rheumatoid arthritis: validation of a gastrointestinal symptom questionnaire. AB - OBJECTIVES: The objectives of this study were to perform an initial validation of the Gastrointestinal Symptom Scale for Kids (GISSK) in children with juvenile rheumatoid arthritis (JRA); and too evaluate the relationship between gastrointestinal (GI) symptoms and health-related quality of life (HRQOL) in JRA. METHODS: A convenience sample of 77 children (median age, 10 years; range, 2-18 years) with JRA requiring second-line agents and one of their parents were interviewed. GI symptoms during the preceding 1 week were measured using the GISSK, which consists of 2 components, a visual analog scale of GI symptom severity (GISSK-VAS) and an 8-item questionnaire (GISSK-Q; score 0-8; 0= no GI symptoms). Information on medications, joint involvement with arthritis, and a physician rating of disease activity were obtained. Patient-centered outcomes included the Childhood Health Assessment Questionnaire (CHAQ) to assess disability and discomfort. HRQOL was measured by the Pediatric Quality of Life Generic Core Scale (PedsQL-GC) and the Rheumatology Module (PedsQL-RM), as well as a visual analog scale (VAS-health). To determine test-retest reliability, the GISSK was completed by 40 parents twice within a 1- to 2-week period. To determine the quality of parent proxy-reporting, parent ratings were compared with those of their children aged 8 years or older. RESULTS: GI symptoms were present in the majority of the patients with JRA (58%). Compared with other patients with JRA, those with a GISSK-Q score of > or =2 had significantly lower HRQOL (PedsQL-GC: P < 0.04; PedsQL-RM: P < 0.05; VAS-health: P < 0.02) and more disability (CHAQ: P < 0.002), despite similar disease activity and joint findings. Similar relationships were observed for the GISSK-VAS with traditional outcomes and HRQOL. The test-retest reliability of the GISSK was good (ICCGISSK-Q = 0.60; ICCGISSK-VAS = 0.67). The quality of parent proxy-reporting was fair to good (ICCGISSK-Q = 0.47; ICCGISSK-VAS = 0.66). CONCLUSION: GI symptoms are frequent among children with JRA requiring advanced therapies and, if moderate or severe, are associated with significantly lower HRQOL. The GISSK is a reliable and valid measure of GI symptoms and their severity in JRA. This self administered measure can be used to screen for GI symptoms in clinical practice and may be useful to assess the effects of medication changes on the perceived GI side effects in children with JRA. PMID- 16357758 TI - Erythromelalgia with subsequent digital necrosis, glomerulonephritis, and antiphospholipid antibodies. AB - Erythromelalgia is a condition of extremities characterized by redness, increased temperature, and burning pain. We describe the first reported case of erythromelalgia in a young woman with digital necrosis and mesangial nephritis associated with antiphospholipid antibodies. The symptoms and necrosis completely resolved with treatment with corticosteroids. We discuss the differential diagnosis of this entity and highlight the importance of a follow up of these patients, because erythromelalgia may precede a myeloproliferative disorder or systemic lupus erythematosus by months or years. PMID- 16357759 TI - Infliximab-responsive uveitis and vasculitis in a patient with Takayasu arteritis. AB - Uveitis rarely occurs in patients with Takayasu arteritis (TA) and may be the presenting manifestation in some. We present the case of a 22-year-old black woman who presented with bilateral uveitis. She was subsequently diagnosed with TA and treated with high doses of corticosteroids and methotrexate. However, she experienced recurrent episodes of uveitis and arteritis on this regimen. Addition of infliximab resulted in resolution of both uveitis and arteritis, thereby permitting tapering of steroids and discontinuation of methotrexate over a period of 1 year. The case highlights the importance of keeping TA in the differential in patients with uveitis and considering infliximab in resistant uveitis and patients with TA. PMID- 16357760 TI - Disseminated giant cell arteritis with inflammatory arthritis and C-ANCA. AB - Giant cell arteritis (GCA) is well known for its involvement of the proximal aorta and its branches, classically causing headache, visual impairment, and elevations in the erythrocyte sedimentation rate (ESR) or C-reactive protein (CRP). We describe a case of biopsy-proven GCA initially presenting with limb claudication, oligoarticular inflammatory arthritis, and a positive antineutrophil cytoplasmic antibody with cytoplasmic staining (C-ANCA), treated successfully with a combination of prednisone and weekly methotrexate. This case illustrates the wide spectrum of features that can be seen with GCA, including the occasional presence of C-ANCA. The C-ANCA became negative after treatment. PMID- 16357761 TI - Successful treatment of childhood onset refractory polyarteritis nodosa with tumor necrosis factor alpha blockade. AB - Polyarteritis nodosa is a rare systemic necrotizing vasculitis of small- and medium-sized arteries that affects patients of all ages. Its incidence ranges from 2 to 9 per million people. The 5-year survival rate is 13% in untreated patients and 77.6% with modern therapy. Standard treatment includes corticosteroids and cyclophosphamide. Despite aggressive medical management, 22.4% of patients die within 5 years, and of the survivors, medication-induced morbidity is frequent. There is great need for better treatment modalities in terms of safety and efficacy. We report the case of a 5-year-old boy with polyarteritis nodosa refractory to all known standard treatments. After 9 years of persistently active disease, at the age of 14, he was successfully managed with the tumor necrosis factor alpha antagonist, etanercept. PMID- 16357762 TI - Acute calcific periarthritis of the hand. PMID- 16357763 TI - Garibaldi's rheumatism. AB - Giuseppe Garibaldi, the charismatic 19th-century Italian revolutionary leader, had complaints of recurrent and often persistent rheumatic pain starting in his early 40s. Symptoms continued until his death at 74, with progressive disability. His diagnosis is uncertain, but rheumatoid arthritis is most likely. Garibaldi coped successfully with his rheumatic illness during most of his military and political career, but eventually became bedridden and dependent. PMID- 16357764 TI - Why I do research. PMID- 16357766 TI - Basic science for the clinician 30: The immunologic synapse. AB - Interactions between discrete and independent cells, immune or otherwise, present a variety of potential problems. How do you make sure the cells can communicate effectively? How do you preclude neighboring cells from "listening in on a conversation" that may be meant only for one set of "ears"? How can you sharpen the hearing of those ears so they will be capable of detecting small signals but not get distracted by random noise in the system? How can you selectively enhance the sense of hearing in times of great need or urgency and then diminish the "gain" of the system when it is not immediately required? How can you assure that the call will be terminated at the end of the conversation? Passage of communication molecules and/or interaction of cell surface markers require close and stable apposition of the cell delivering the message with the receiving cell. In the nervous system, these problems were successfully addressed in the nerve nerve or nerve-myocyte (neuromuscular junction) synapses. Not surprisingly, given the parsimony of nature, the immune system uses some of the same design features, even some of the same molecules, to achieve an effective communication strategy. The term "immunologic synapse" was coined only 2 decades ago, but the structure it describes has become a very hot topic in immunology and cell biology. The immunologic synapse allows the activation of a unique T cell, with an antigen receptor recognizing its antigen in the grasp of the antigen-presenting cell's (APC's) major histocompatibility complex (MHC). A better understanding of this transient immune cell-cell interactive structure allows one to weave the functions of T cell antigen receptors, lipid rafts, adaptor molecules, and nuclear signaling molecules together into one cohesive, flowing communication supersystem. Appreciation of the intricacies of the synapse also identifies targets that one day may be used to interfere with antigen-specific immune responses, eg, autoimmunity. PMID- 16357767 TI - Acute gouty arthritis is seasonal: possible clues to understanding the pathogenesis of gouty arthritis. PMID- 16357768 TI - First-time manifestation of giant-cell arteritis during methotrexate treatment. PMID- 16357769 TI - Temporal arteritis presenting with paresis of the oculomotor nerve, and polymyalgia rheumatica, despite a low erythrocyte sedimentation rate. PMID- 16357770 TI - Absolute reduction in lower limb lean body mass in Japanese women with knee osteoarthritis. AB - BACKGROUND: Knee osteoarthritis (OA) is a significant contributor to disability, and obesity is one of the most modifiable risk factors. The association between knee OA and obesity has been reported to relate to quadriceps strength. To begin to examine this hypothesis, independent measures of quadriceps lean mass are needed. OBJECTIVE: The objective of this study was to characterize the absolute lower limb lean body mass in women with knee OA. METHODS: Japanese women, aged 45 and over, who presented to a community orthopedics clinic, were categorized as 1) those with no knee pain, presenting with fracture, sprain, or back pain (n = 604); and 2) those presenting with knee pain meeting clinical and radiographic criteria for knee OA (n = 341). Segmental lower limb lean body mass (L-LBM) was measured with multifrequency bioelectrical impedance analysis and subjects were stratified by body mass index (BMI) by Western Pacific Region World Health Organization weight categories. RESULTS: Control and knee OA groups were comparable with respect to age and BMI. Despite an increase in L-LBM with increasing BMI, there was a statistically significant reduction in L-LBM in knee OA subjects in comparison with control subjects in matched normal weight, overweight, and obese strata (11.1, 12.4, and 13.1 kg in the nonknee OA compared with 10.5, 10.6, and 11.6 kg in the knee OA group). CONCLUSIONS: A reduction in absolute L-LBM is observed in older Japanese women with knee OA in comparison with control subjects. Longitudinal studies are warranted in which L-LBM can be prospectively evaluated in a cohort followed for incident knee OA to better characterize the temporal relationship with L-LBM decline. Clinicians should consider this reduction in L-LBM in women with knee OA to recommend therapy aimed at reducing risk for atrophy and disability. PMID- 16357771 TI - Neuropsychologic functioning and health status in systemic lupus erythematosus: does ethnicity matter? AB - BACKGROUND: Despite increased severity of lupus in blacks, including more frequent neuropsychiatric manifestations, there is sparse data on neuropsychologic function in black patients with lupus. METHODS: Neuropsychologic functioning and health-related variables were examined among blacks (n = 34) and whites (n = 14) fulfilling American College of Rheumatology criteria for systemic lupus erythematosus. RESULTS: Blacks and whites performed comparably on measures of verbal and visual memory, working memory, and motor speed after controlling for estimates of premorbid cognitive ability. Blacks trended towards poorer performance on specific attention/processing speed measures. Pain, fatigue, depression, anxiety, physical and emotional well-being were unrelated to ethnicity. Blacks exhibited a trend towards greater impairment of physical functioning. Ethnicity-related differences in overall damage, noncognitive neuropsychiatric manifestations, and prevalence of nephritis revealed greater severity among blacks. CONCLUSIONS: Initial differences in premorbid cognitive function possibly contribute to disparate clinical outcomes, including a greater proportion of blacks exhibiting subnormal neurocognitive performance. Blacks evidencing lower premorbid ability may be at greater vulnerability for poorer functional outcomes (eg, coping skills, medical compliance and employment) if they experience disease-related cognitive dysfunction. PMID- 16357772 TI - Successful treatment of postvenereal reactive arthritis with synovectomy and 3 months' azithromycin. AB - BACKGROUND: The effects of antibiotic therapy on the course of postvenereal reactive arthritis have not yet been elucidated. OBJECTIVE: The objective of this study was to observe the clinical course and outcome of synovectomy and 3 months of azithromycin therapy in patients with reactive arthritis and previously diagnosed triggering bacteria. METHODS: We performed an open, prospective study on 20 (14 male/6 female) patients with postvenereal reactive knee arthritis, aged 36.7 +/- 14.8 years, and with 16.5 +/- 20.4 months' duration of the disease. Detection of bacteria triggers was done by polymerase chain reaction, isolation and identification, and electron microscopy. Synovectomy was performed in all patients at entry into the study. Azithromycin was given at a dose of 500 mg per day for 5 days, and then 500 mg twice a week, during a 3-month period. Patients without remission were treated with combined antibiotic therapy using a macrolide, quinolone, and tetracycline for the next 4 months. Outcome evaluations of therapeutic efficacy and azithromycin safety were done after 1 and 3 months and 2 years of follow up. RESULTS: Remission, defined by the absence of joint swelling and tenderness, and extraarticular signs, was reached after 3 months in 15 of 20 (75.0%) patients (P = 0.025). Of 5 patients with persistent knee arthritis, remission was achieved with combined antibiotic therapy in 4. Visual analog scale scores (P < 0.01), the number of patients (P = 0.002), and the number of samples (P = 0.01) with a positive finding of bacteria or their DNA were significantly lower after 3 months of therapy. During the azithromycin therapy, there were no significant adverse effects. CONCLUSIONS: These patients with reactive arthritis did extremely well on the regimen described. In our study group, the number of patients and the number of samples with positive findings of bacteria or their DNA were lower after the antibiotic treatment combined with surgery, although not all bacteria were eradicated. Adverse effects of prolonged azithromycin administration were insignificant. This open treatment approach is recommended but does need a study with controls. PMID- 16357773 TI - Rituximab in the treatment of refractory dermatomyositis. AB - Dermatomyositis is an inflammatory myopathy characterized by muscle weakness and inflammation. In contrast to polymyositis and inclusion body myositis, humoral immune mechanisms appear to contribute to the pathogenesis of dermatomyositis. A 56-year-old man with dermatomyositis resistant to conventional therapies was treated with 6 weekly infusions of the anti-CD-20 monoclonal antibody, rituximab, at a dosage of 100 mg/m in addition to other agents. The patient demonstrated a remarkable clinical response as indicated by an increase in muscle strength and a decline in creatine kinase enzymes. B-cell depletion therapy with rituximab used alone or in combination with other immunosuppressive therapies may be a viable option in patients with dermatomyositis as well as other autoimmune diseases refractory to current therapies. PMID- 16357774 TI - Wegener granulomatosis presenting as bilateral loss of vision. AB - We describe a man presenting with bilateral loss of vision secondary to a branch retinal artery occlusion and ischemic optic neuropathy, in turn related to underlying Wegener granulomatosis (WG). This case is unusual, because the simultaneous development of branch retinal artery occlusion and anterior ischemic optic neuropathy has, to our knowledge, not been reported in WG. Furthermore, only rarely does WG cause bilateral vision loss. The patient responded to immunosuppressive therapy and recovered sufficient vision to resume his vocation as a farmer. PMID- 16357775 TI - Reactive thrombocytosis: an early manifestation of Takayasu arteritis. AB - A woman with Takayasu arteritis is reported who presented with constitutional symptoms and persistent thrombocytosis documented since 3 years before the diagnosis. Disease-specific symptoms such as arm claudication, transient loss of vision, and self-remitting eye ptosis present at the time were apparently missed, because she is a non-English-speaking Hispanic woman whose history was obtained through an interpreter. Extensive workup done at the time failed to reach a definite etiology. A computed tomography scan of the chest done because of midthoracic back pain and an elevated erythrocyte sedimentation rate showed circumferential wall thickening and mild surrounding edema throughout the thoracic and abdominal aorta and both carotid arteries consistent with Takayasu arteritis. Prednisone at a dosage of 1 mg/kg twice a day decreased the platelet count within 45 days of its initiation. Takayasu arteritis should be considered in the differential diagnosis of unexplained thrombocytosis, particularly in young women. PMID- 16357776 TI - Musician's cramp: a case report and literature review. AB - Musician's focal dystonia is a motor dysfunction that appears in artists after years of repetitive and fine movements during performance. This is the condition most feared by musicians because it leads to difficulties in controlling movements, which can interrupt or terminate their careers. It is characterized by the onset of involuntary muscle contractions and movements; its distinguishing feature is that it only occurs during a specific and well-defined action. It is rarely diagnosed because those who experience it do not seek medical attention for fear or shame, but also because many physicians do not consider the disease in the differential diagnosis of motor dysfunction. We describe the case of a guitarist who presented to our outpatient clinic after many years of experiencing musician's focal dystonia. He reported a long list of misdiagnoses and a variety of unsuccessful treatments.Musician's focal dystonia is an under-diagnosed condition. Treatment benefit is limited despite recent innovative approaches. Rheumatologist should be aware of this condition. PMID- 16357777 TI - Suspected gouty tophus in a woman's breast. AB - A postmenopausal woman treated with diuretics for hypertension and who underwent surgery for a previous fibroadenoma of the left breast presented, in December 2003, with a sudden bloody discharge of the left nipple with local swelling, redness, and pain. Radiologic investigations revealed a well-delimited tumor under the areola. Needle aspiration disclosed many histiocytes but no bacteria. Despite improvement with antibiotics, the tumor was removed. Pathological examination shows features suggesting the tumor was a gouty tophus. Birefringent crystals were retrospectively found on the smears of the aspirated material. To our knowledge, only 3 similar cases have been reported in the literature. PMID- 16357778 TI - Osteomyelitis resulting from chronic filamentous fungus olecranon bursitis. AB - We describe a case of Phaeoacremonium olecranon osteomyelitis. The patient, initially felt to have traumatic olecranon bursitis, was found to have an indolent filamentous fungus cultured from the olecranon bursa. In retrospect, x rays revealed bony erosion, which heightened the index of suspicion for infection in this particular case. Surgical bursal excision was performed and antifungal therapy was administered with clinical resolution. This case emphasizes that aspiration, synovial fluid analysis, and culture of bursal fluid is essential in excluding typical and atypical causes of chronic bursitis. PMID- 16357779 TI - Vertebral anomalies associated with Goldenhar syndrome. PMID- 16357781 TI - Basic science for the clinician 32: T-cells with regulatory function. AB - Tolerance is one of the major requirements of a successful immune system: destroy invaders but recognize self and the benign environment to leave them alone. Central tolerance is achieved by deletion of T-cells with T-cell antigen receptors that recognize self-antigens too well. However, within the population of T-cells that actually survive the harrowing experience of passage through the thymus (wherein over 98% of all pre-T-cells perish), there persist T-cells capable of inducing autoimmune damage. How then to avoid autoaggression? One theory in the 1970s was that there were peripheral T-suppressor cells that actively dampened these autoimmune proclivities. One problem presented itself, however; despite the fact that such an immunologic activity could be measured, no one could identify the cells that mediated the activity and so the concept fell into disfavor. However, the concept of peripheral regulation by a distinct (at last identifiable!) population of T-cells is now back in vogue. These T regulatory (T-reg) cells have an important function in immune homeostasis. T-reg demand our attention as we try to manipulate the molecular biology of immune responses and inflammation to control autoimmune disorders and enhance transplantation efficiency. PMID- 16357782 TI - Occurrence of vesicocolic fistula with a use of etanercept: a cautionary tale. PMID- 16357783 TI - Complete long-term serologic and clinical remission in a patient with severe systemic lupus erythematosus. PMID- 16357784 TI - New-onset lupus presenting as serositis in an 80-year-old woman: does a high titer ANA in pleural, pericardial, or peritoneal fluid help confirm the diagnosis? PMID- 16357785 TI - Is non-insulin dependent glucose uptake a therapeutic alternative? Part 1: physiology, mechanisms and role of non insulin-dependent glucose uptake in type 2 diabetes. AB - Several decades of research for treating type 2 diabetes have yielded new drugs but the actual experience with the available oral antidiabetic compounds clearly shows that therapeutic needs are not matched. This highlights the urgent need for exploring other pathways. All cell types have the capacity to take up glucose independently of insulin, whereby basal but also hyperglycaemia-promoted glucose supply is ensured. Although poorly explored, insulin-independent glucose uptake might nevertheless represent a therapeutic target, as an alternative to the clear limits of actual drug treatments. This review not only critically examines some major pathways not requiring insulin (although they may be influenced by the hormone) but importantly, this analysis extends to the clinical applicability of these potential therapeutic principles by also considering their predictable tolerability for long-term therapy. In particular vascular safety (the ultimate problem linked with diabetes) will be envisaged because of the ubiquitous distribution of glucose transporters and some linked mechanisms. Several mechanisms can be identified which do not require insulin for their functioning. The first part of this review deals with the description, the regulation and the limits of some mechanisms representing potential pharmacological targets capable of having a highly significant impact on glucose uptake. These selected topics are: a) unmasking and/or activation of glucose transporters in cell plasma membranes, b) insulin mimetics acting at postreceptor level, c) activation of AMPK, d) increasing nitric oxide and e) increasing glucose-6P and glycogen stores. PMID- 16357786 TI - New insight into the pathophysiology of lipid abnormalities in type 2 diabetes. AB - Lipid abnormalities in patients with type 2 diabetes are likely to play an important role in the development of atherogenesis. These lipid disorders include not only quantitative but also qualitative abnormalities of lipoproteins which are potentially atherogenic. The main quantitative abnormalities are increased triglyceride levels, related to an augmented hepatic production of VLDL and a reduction of both VLDL and IDL catabolism, and decreased HDL-Cholesterol levels due to an accelerated HDL catabolism. The main qualitative abnormalities include large VLDL particles (VLDL1), relatively rich in triglycerides, small dense LDL particles, increase in triglyceride content of LDL and HDL, glycation of apolipoproteins and increased susceptibility of LDL to oxidation. Moreover, although plasma LDL-cholesterol level is usually normal in type 2 diabetic patients, LDL particles show significant kinetic abnormalities, such as reduced turn-over, which is potentially harmful. The pathophysiology of lipid abnormalities in type 2 diabetes is not yet totally explained. However, insulin resistance and the "relative" insulin deficiency, observed in patients with type 2 diabetes, are likely to play a crucial role since insulin has an important function in the regulation of lipid metabolism. In addition, it is not excluded that adipocytokines, such as adiponectin, could play a role in the pathophysiology of lipid abnormalities in type 2 diabetes. PMID- 16357787 TI - Knowledge about preconception care in French women with type 1 diabetes. AB - OBJECTIVE: To assess the knowledge about preconception care among women with type 1 diabetes of childbearing age and to identify factors that may be associated with the absence of preconception care. RESEARCH DESIGN AND METHODS: All women of childbearing age with type 1 diabetes who were seen in 11 Diabetes Centers between December 1 and 12, 2003 were included. An anonymous dual questionnaire was proposed to these women and their consulting diabetologists. RESULTS: One hundred and thirty-eight women were included in the study. The main reported sources of information about pregnancy were the diabetologist (78%) and/or specific leaflets (42%). Although 85% of the women declared having received information about preconception care, 48% were unaware of the risk of congenital malformations and 41% feared for neonatal diabetes. However, 82% of the women thought that a level of HbA1c below 7% was a target to achieve before conception. In multivariate analysis, onset of diabetes before the age of 15 was associated with the knowledge that diabetes may be associated with complications during pregnancy; the use of contraception was associated with knowledge about preconception care; a high educational level was associated with knowledge that complications are avoidable and that good glycemic control is mandatory before conception. CONCLUSIONS: French women with type 1 diabetes, although followed by diabetologists, have major knowledge defects concerning the risks associated with pregnancy. This is the first step towards decreasing the rate of unplanned pregnancies in women with type 1 diabetes. PMID- 16357788 TI - Major lower limb amputations in the elderly observed over ten years: the role of diabetes and peripheral arterial disease. AB - BACKGROUND: Major amputation is a dreaded event with high mortality and morbidity. However, few studies have investigated the epidemiology of amputation in the elderly over time, in the face of evolving management and prevention efforts. METHODS: We undertook a retrospective study to determine the incidence rate, etiology and prognosis of major lower limb amputations (transtibial or higher) in elderly patients (> 65 years). Cases were identified over a 10-year period in the Geneva (Switzerland) area, where all amputations are performed in a single center and reliable demographic data are available. RESULTS: The rate of amputation varied from 1.8 to 11.4/10000 patients/year, increasing with age and male gender. Diabetes was present in 48% patients, and conferred a 10 times higher risk of amputation. Severe peripheral arterial disease (PAD) was present in > 94% patients. The prognosis remains poor, 47% patients had died after two years and only 53% patients could be equipped with a prosthetic limb. Over 10 years we found a progressive increase in age at amputation; this encouraging increase was mostly accounted for by diabetic patients (> 6 months per year). CONCLUSIONS: The rate of amputation observed among elderly patients was low. Neither the rate nor the prognosis improved over the decade studied. However, the age at amputation increased by > 6 months/year, particularly in diabetic amputees, suggesting that current management successfully delays amputation. Amputations were almost exclusively performed for severe PAD. Further reduction in the rate of amputation will require progress in the prevention and management of PAD. PMID- 16357789 TI - Patterns of prescription of hypoglycaemic drugs in Gran Canaria (Canary islands, Spain) and estimation of the prevalence of diabetes mellitus. AB - OBJECTIVE: To study the patterns of hypoglycaemic treatment in our community and to estimate the prevalence of known and drug-treated diabetes mellitus. METHODS: From all the diabetic patients who attended the Healthcare Centers of the National Health Service in Gran Canaria in 1999, a random sample of 2924 diabetic patients > 20 years old was selected. Data on age, gender, clinical onset of diabetes, and hypoglycaemic treatment were obtained. Data on drug consumption were supplied by the National Health Service. RESULTS: Of the DM-2 patients 4.4% (3.65-5.14) 84.2% (82.7785.42), 9.4% (8.34-10.45) and 2.1% (1.58-2.61) received diet only, oral drugs, insulin or combination. The duration of DM-2 was associated with more oral drugs and more insulin treatment, but the duration of DM-1 was not associated with intensive insulin therapy;<50% of the type 1 patients had >or=3 daily injections. The prescriptions of biguanides were scarce; over 1/3 of them were of buformin. DM-1 and DM-2 patients were treated with similar doses of insulin, but DM-1 patients had more insulin injections (2.56 vs 2.07, P<0.001), and more fast-acting insulins (65.2% vs 38.0%, P<0.001). The estimated prevalences of known and drug-treated diabetes in the Gran Canaria island were 5.95% (95% CI: 5.096.80%) and 5.73% (4.88-6.57%). CONCLUSIONS: Our prevalences of known and drug-treated diabetes is among the highest reported in European populations. The prescriptions of metformin and of combined therapy in DM-2, and of intensive insulin therapy in DM-1 are less frequent than expected, but nonetheless insulin therapy in DM-1 is more intensive and uses more fast acting insulin than in DM-2. PMID- 16357790 TI - Metabolic factors and the foveal avascular zone of the retina in diabetes mellitus. AB - AIM: To study the foveal avascular zone (FAZ) of the central retina in diabetic patients with retinopathy having undergone metabolic evaluation. METHODS: One hundred and ten digital fluorescein angiograms were chosen from our digital image bank after cross matching diabetic patient lists of the ophthalmology and endocrinology departments of our institution. The patients had undergone day visits with systemic, biological and ophthalmologic evaluation, including digital fluorescein angiography. RESULTS: Sex ratio was M 62/F 48. Average age was 52.4 years (+/- 13.8) with 44 type 1 diabetics and 66 type 2. Retinopathy was present in all patients (54 background (BDR), 30 pre-proliferative (PPDR), 26 proliferative (PDR)). Age was positively correlated with FAZ grade (47.3 years +/ 13.2 for normal FAZ, 53.8 years +/- 13.7 for abnormal FAZ, P=0.03). Lipid profile showed a protective tendency of the Apo A1 fraction of cholesterol on macular vascularization (1.7 gr./l in normal FAZ patients vs 1.43 gr./l in abnormal FAZ patients, P=0.004). Body mass index was negatively correlated with macular ischemia (28.11 if FAZ not severely altered, 25.97 if FAZ severely altered, P=0.03). CONCLUSIONS: We found possible relations between BMI and Apo A 1 cholesterol and macular vascularization which may warrant further investigation. PMID- 16357791 TI - Abdominal fat index by ultrasound does not estimate the metabolic risk factors of cardiovascular disease better than waist circumference in severe obesity. AB - OBJECTIVE: To evaluate by ultrasound the ratio between preperitoneal (P) and subcutaneous (S) fat (AFI), in quantifying the cardiovascular risk in 258 obese patients (BMI 41.2+/-6.3 kg/m2; age 45.1 +/- 13.6 years). RESEARCH METHODS AND PROCEDURES: Glucose, insulin, lipid profile, uric acid and fibrinogen were measured. HOMA-IR, waist girth, AFI and quartiles of BMI were calculated. RESULTS: AFI lowered with increasing BMI and showed a positive correlation with TGL (r=0.37, P<0.01) and uric acid (r=0.40, P<0.001) in the 1st quartile of BMI (30.2-36.4) and a negative correlation with HDL (r=- 0.32, P<0.001) in the 3rd quartile (40.6-45.1). When BMI exceeded the value of 45.2 kg/m2 these correlations were no longer significant. In all subjects S correlated positively with uric acid (r=0.64, P<0.001), and negatively with HOMA-IR (r=- 0.41, P<0.001) and TGL (r=- 0.35, P=0.02); P correlated positively with CHOL (r=0.48, P=0.04) and TGL (r=0.33, P=0.03), and negatively with HDL (r=- 0.46, P=0.03). Waist girth showed more significant correlations than AFI in the lower quartiles of BMI, but not at the highest one. DISCUSSION: AFI, P and S, as waist girth do not seem to quantify the metabolic risk factors of cardiovascular disease in severe obese subjects, but AFI is probably useful in obese populations with BMI<45 kg/m2, even though not as strong as waist girth. PMID- 16357792 TI - Control of cardiovascular risk factors in patients with type 2 diabetes and hypertension in a French national study (Phenomen). AB - For some years now, the recommendations of scientific societies have significantly reduced the therapeutic targets for blood pressure, glycaemia and lipid levels in diabetic patients. However, little is known regarding the synchronization between effective risk factor management and the guidelines. To examine this issue, the Phenomen survey was conducted between January and July 2001 on 16358 patients suffering from hypertension followed by a general practitioner in France. AIM: To evaluate the control of cardiovascular risk factors in patients with diabetes and hypertension according to the French guidelines. METHODS AND PATIENTS: 8177 general practitioners, selected from a national database according to quotas, taking into account age, practice and area, had to include the first two hypertensive patients they came across in their practice and to collect their demographic data, cardiovascular risk factors and medications. RESULTS: 2346 out of 16358 hypertensive patients presented with type 2 diabetes (14.3% of the cohort). The number of GP consultations in the last 12 months averaged 8.31. According to the French guidelines, 6.5% had a blood pressure<140/80 mmHg, a total of 38.7% patients met the goal of LDL cholesterol level and 26.6% of patients had an HbA1c<6.5%, 53.4% of patients had an HbA1c between 6.6 and 8%. 37.1% of patients continued to receive antihypertensive monotherapy but only 3% in this monotherapy group reached the target of 130/85 mmHg. 29% of the patients were on antiplatelet therapy. 64.6% of these hypertensive diabetic patients presented with more than three other cardiovascular risk factors. Based on WHO recommendations, 0.3% of the patients met all of the blood pressure, lipid and glycaemic treatment objectives. CONCLUSION: Despite frequent monitoring by a general practitioner, the overall management of modifiable risk factors in this diabetic hypertensive population is clearly inadequate. The impact of the guidelines on effective management remains limited and additional information is required to understand why physicians are not more aggressive in managing modifiable risk factors in diabetic patients. PMID- 16357793 TI - Blood pressure fall and increased relaxation of aortic smooth muscle in diabetic rats. AB - This study was designed to identify changes in endothelium-independent relaxation that could contribute to the depressed vascular reactivity and fall in blood pressure (BP) detected in rats after 5 weeks of streptozotocin (STZ)-induced (i.e. type 1) diabetes. Aortic rings were contracted by simultaneous activation of voltage-operated channels (KCl=80 mM) and alpha-adrenergic receptors (phenylephrine 1 microM) and then relaxed by simultaneous exposure to Ca2+-free PSS and 10 microM phentolamine. Additional relaxations were performed under conditions in which the plasma membrane Na-Ca exchanger (PMNaCa) or Ca-pump (PMCA), or the sarcoplasmic reticulum (SR) Ca-pump (SERCA) were blocked, to identify which mechanism(s) could modulate this process. The STZ-diabetic rats had a moderate but significant decrease of BP, and their aortic rings exhibited accelerated relaxation following a biexponential model, with a significantly decreased slow component. In control rats only the inhibition of the PMNaCa could slow down the fast component, while the slow component was insensitive to any blocking maneuver. In contrast, the diabetic animals' slow component was sensitive to the inhibition of both the PMNaCa and the SERCA. The SERCA-sensitive 45Ca2+ uptake by the SR was augmented in the aortas of STZ-treated animals. This hyperactivity of the SERCA, associated with augmented activity of the PMNaCa, at least partly induced by an increase of the plasma membrane Na+/K+-ATPase activity, could explain the decrease in BP and the accelerated aortic relaxation observed in the diabetic rats. PMID- 16357794 TI - Subcutaneous insulin resistance successfully circumvented on long term by peritoneal insulin delivery from an implantable pump in four diabetic patients. AB - Extreme subcutaneous insulin resistance is a rare syndrome characterized by a severe resistance to subcutaneous (S/C) insulin together with persistence of normal or near normal intravenous (IV) insulin sensitivity. Its pathophysiology is unknown, although increased insulin degrading activity has been reported in the S/C adipose tissue fraction in some cases. Until now, proposed treatments have been disappointing. We report 4 cases who were successfully treated by intraperitoneal (IP) route. METHODS: The diagnosis of subcutaneous insulin resistance was based upon following combined conditions: resistance to hypoglycaemic action of subcutaneous insulin but normal or near normal sensitivity to IV or IP insulin. RESULTS: 4 patients among those followed by EVADIAC group met these criteria: 3 with type 1 diabetes (C peptide=0), the last one with unexplained non insulin-deficient diabetes (no anti-GAD antibodies, C peptide=5 ng/ml). All of them had been treated with subcutaneous insulin therapy without success despite huge doses (up to 4000 IU/day in two patients). The 3 type 1 diabetic patients presented with a history of repeated ketoacidosis episodes. A treatment of insulin mixed with aprotinin had been proposed to 2 patients without success. The IV insulin sensitivity was proved to be normal in two patients by euglycaemic clamp data. A skin biopsy was performed in 1 patient. An accumulation of insulin in the derma was revealed with no increase of degradation products of insulin. In these 4 patients, a dramatic improvement of diabetes control was obtained by IP insulin delivery from an implantable pump (HbA1c decrease by at least 3%). CONCLUSION: Although pathophysiology of the subcutaneous insulin resistance syndrome remains unexplained, our data show that intra-peritoneal insulin therapy from an implantable pump allows diabetes control in patients affected by this uncommon but severely disabling condition. PMID- 16357795 TI - Changes in diabetes prevalence and treatment in the last ten years in Luxembourg. A lesson from the United Kingdom prospective diabetes study? AB - OBJECTIVES: The aims of the study were to estimate the prevalence of diabetes in Luxembourg in 2002, to compare it to the prevalence reported in 1991 and to evaluate if prescription attitudes have changed since 1991. METHODS: The prevalence of diabetes was estimated using the drug sales data. The key parameters, total amount of antidiabetic drugs sold in one year and the average daily dose or Prescribed Daily Dose (PDD), have been obtained from the National Social Security Organization and by a standardized questionnaire sent to all general practitioners and all internists and endocrinologists of the country. RESULTS: The PDD was calculated on 2, 402 questionnaires on individual diabetic patients. By this means, the proportion of patients only treated with appropriate diet could also be obtained. Compared to 1991, the total amount of antidiabetic drugs showed a four-fold increase in metformine tablet prescriptions. A high percentage of combined treatments was found. The prevalence of diabetes in Luxembourg was found to be 3.05% of the total population. CONCLUSIONS: Compared to the status in 1991, prevalence of diabetes increased by 63%, which seems mainly due to type 2 diabetic patients as orally-treated diabetic patients almost doubled (2.11% vs 1.16%). A substantial change in prescriptions for diabetes has occurred, suggesting a positive influence of studies like the United Kingdom Prospective Diabetes Study (UKPDS). PMID- 16357796 TI - Cardiovascular risks in type 2 diabetes and secondary cardiovascular prevention. AB - In order to emphasize the necessity of treating aggressively the main cardiovascular risk factors (hypertension, hypercholesterolemia, smoking) in diabetics, it was proposed in 1998 to assimilate their absolute risk level to that of non diabetics with a history of myocardial infarction. In this short review, it is recalled that diabetics (and coronary patients as well) constitute heterogeneous groups and the results of such comparisons depend heavily on the selection of the two groups as shown by published works on this topic since 1998. Anyway, the treatment of the main risk factors in true diabetics should be strict and new preventive approaches by treating the disease itself are awaited. PMID- 16357798 TI - Prediabetes or what's in a name? PMID- 16357799 TI - Is non-insulin dependent glucose uptake a therapeutic alternative? Part 2: Do such mechanisms fulfil the required combination of power and tolerability? AB - The worldwide burden of diabetes, the unavoidable worsening which is observed in long-term clinical trials despite treatment and the close link between glycaemia and microangiopathy appeal for much stronger treatment strategies. This, in turn, either requires polypharmacy (with new risks) or new, more powerful drugs to be invented. The first part of this review dealt with a thorough analysis of pros and cons for some selected pathways which could potentially increase glucose uptake without necessitating insulin. The choice of such targets for developing completely new drugs, however, requires a favourable background from existing tentatives with either drugs or cell biology approaches. Moreover, because vascular complications are what must ultimately be avoided when treating diabetic patients, we must be sure that increasing glucose uptake in a fashion which is no more controlled by normal physiology is compatible with the physiology of vascular cells (long-term tolerance). The aspect of drug side-effects must therefore be considered systematically. For reasons which are individually developed, it appears that each of the potential pathways analyzed either lacks sufficient power and/or is likely to induce side effects which are not acceptable for long-term application. The fact that GLUT-1 transporters are ubiquitously distributed even extends this cardinal question to the general principle of increasing glucose uptake. In conclusion a precise evaluation suggests that, although non-insulin dependent glucose uptake represents (3/4) of whole body glucose transport, it is difficult to consider such mechanisms able to generate a new treatment fulfilling the unavoidable request of combined efficacy and tolerability. PMID- 16357800 TI - Etiological investigations in apparent type 2 diabetes: when to search for lamin A/C mutations? AB - Prevalence of diabetes is increasing worldwide in epidemic proportions. Its appropriate clinical management requires a careful etiological diagnosis. Laminopathies recently emerged as clinically heterogeneous genetic disorders due to mutations in lamins or lamin-associated proteins, which are components of the nuclear envelope. Laminopathies regroup at least eight distinct diseases, belonging to the groups of skeletal and/or cardiac muscular dystrophies, axonal neuropathies, premature ageing syndromes and familial lipodystrophies, all resulting from alterations in LMNA, encoding type A-lamins. Pathophysiological mechanisms explaining how mutations in an unique gene could lead to such various phenotypes are still unknown, but probably involve alterations in cellular mechanical stress responses, in gene expression, and/or in post-translational maturation of lamin A. Familial Partial Lipodystrophy of the Dunnigan type (FPLD2), with specific features of pseudo-cushingoid lipodystrophy, marked insulin resistance and muscular hypertrophy, and a relatively homogeneous genotype, was thought, until recently, to be the only laminopathy causing diabetes. However, recent studies have revealed that insulin resistance and diabetes could be key features of attenuated or more complex phenotypes of laminopathy. In the light of these recent findings, this review will describe the clinical, morphological and biological features that should lead clinicians to consider the diagnosis of laminopathy in a diabetic patient. The recognition of such an etiology for diabetes is important not only for its appropriate medical treatment, but also because specific investigations are required to detect possible asymptomatic life-threatening complications. In addition, the molecular screening of family members allows an earlier efficient clinical management of affected relatives. PMID- 16357801 TI - Reliability of bioimpedance analysis compared with other adiposity measurements in children: the FLVS II Study. AB - OBJECTIVE: To evaluate the reproducibility of the measurement of% body fat by bipedal biometrical impedance analysis (BIA) compared with anthropometric measurements of adiposity in children and the correlations between these methods in children and adults. METHODS: A cross-sectional study in a total of 1080 adults and children enrolled in 1999 in the Fleurbaix-Laventie Ville Sante II (FLVS II) population-based study in northern France. The reproducibility of anthropometrical and BIA methods was determined by a nested analysis of variance of repeated measurements by 2 investigators and a bipedal BIA device (Tanita TBF 310) in 64 pupils of two 5th grade classes. The correlation of BIA and anthropometric adiposity measurements with the unknown relative fat mass or volume of the body estimated by a latent adiposity variable (LAV) was established by the triads' method in 1080 subjects of the FLVS II cohort. RESULTS: The reproducibility was similar for the sum of skinfolds, waist circumference and BIA% fat measurements (intraclass correlation coefficients: 0.979-0.992). Correlation coefficient between BIA body fat% and the LAV was higher than 0.86 in all sex and Tanner stage related groups, and similar in children and adults, except in pubertal boys (0.76). CONCLUSION: With a high level of reproducibility, foot-to-foot BIA analysis provides a valuable measurement of total% fat for epidemiologic studies in children. However further studies are needed before extrapolating these results to overweight children. PMID- 16357802 TI - Hepatic markers and development of type 2 diabetes in middle aged men and women: a three-year follow-up study. The D.E.S.I.R. Study (Data from an Epidemiological Study on the Insulin Resistance syndrome). AB - AIM: It has been shown, mainly in men, that gamma-glutamyltransferase (gammaGT) and alanine-aminotransferase (ALT) predict the development of type 2 diabetes. This study investigates the association between hepatic markers and the 3-year risk of diabetes. METHODS: Incident diabetes was studied in 2071 men and 2130 women without diabetes at baseline from the D.E.S.I.R. cohort. RESULTS: Adjusting on age, only gammaGT was predictive of diabetes in both sexes, whereas ALT and aspartate-aminotransferase (AST) were only predictive in men, and bilirubin was not predictive. After adjustment on classical confounding factors and on ALT activity, the odds ratios (OR) for incident diabetes increased across baseline gammaGT quartiles: 1, 3.1, 2.6, 5.0 in men (P<0.0003) and 1, 0.9, 3.2, 3.5 in women (P<0.01). The relations with ALT and AST were not significant after adjusting on gammaGT. Additional adjustment on markers of insulin resistance, BMI or fasting plasma glucose attenuated the risk associated with gammaGT in both sexes, and it remained significantly predictive only in the men. Pooling men and women, those with gammaGT above the median had adjusted ORs of developing diabetes of 13.7 (1.8-99.8) if WHR > =0.85 and 1.7 (0.6-4.8) if WHR<0.85 (interaction P<0.007). CONCLUSION: gammaGT was the main hepatic risk marker for type-2 diabetes in both sexes, especially in subjects with central adiposity. PMID- 16357803 TI - Protein oxidation parameters in type 2 diabetic patients with good and poor glycaemic control. AB - AIM: In order to examine the influence of oxidative stress on protein oxidation, type 2 diabetic patients without clinical evidence of complications, either in good or poor glycaemic control, were studied. METHODS: Plasma protein carbonyl (PCO), total thiol (T-SH), and advanced oxidation protein products (AOPP) levels as markers of protein oxidation, and lipid hydroperoxide (LHP) levels as markers of lipid peroxidation were determined. Glycated haemoglobin (HbA1c) levels were used as an index of glycaemic control. The subjects were divided into two groups according to their HbA1c level at inclusion as follows: good HbA1c<=7%, and poor HbA1c > 7%. RESULTS: Plasma PCO and AOPP levels of diabetic patients with poor glycaemic control were increased significantly compared with those of the diabetic patients with good glycaemic control. The decreased plasma T-SH level in the diabetic patients with poor glycaemic control was not statistically significant. On the other hand, plasma LHP levels were increased significantly in the diabetic patients with poor GC compared with those of the diabetic patients with good glycaemic control. CONCLUSION: This study supports the hypothesis that poor glycaemic control is an important factor in generation of increased protein oxidation in type 2 diabetic patients clinically free of complications. Increase in plasma PCO, AOPP, and LHP levels in the diabetic patients with poor glycaemic control may contribute to the development of diabetic complications. PMID- 16357804 TI - In vivo expression of carbohydrate responsive element binding protein in lean and obese rats. AB - ChREBP (Carbohydrate response element binding protein) is considered to mediate the stimulatory effect of glucose on the expression of lipogenic genes. Its activity is stimulated by glucose. Less is known on the control of its expression. This expression could be controlled by nutritional (glucose, fatty acids) and hormonal (insulin) factors. We examined the in vivo nutritional control of ChREBP expression in liver and adipose tissue of Wistar rats. Compared respectively to the fed state and to a high carbohydrate diet, ChREBP mRNA concentrations were not modified by fasting or a high fat diet in rat liver and adipose tissue. FAS and ACC1 mRNA concentrations were on the contrary decreased as expected by fasting and high fat diets and these variations of FAS and ACC1 mRNA were positively related to those of SREBP-1c mRNA and protein, but not of ChREBP mRNA. Therefore i) ChREBP expression appears poorly responsive to modifications of nutritional condition, ii) modifications of the expression of ChREBP do not seem implicated in the physiological control of lipogenesis. To investigate the possible role of ChREBP in pathological situations we measured its mRNA concentrations in the liver and adipose tissue of obese Zucker rats. ChREBP expression was increased in the liver but not the adipose tissue of obese rats compared to their lean littermates. These results support a role of ChREBP in the development of hepatic steatosis and hypertriglyceridemia but not of obesity in this experimental model. PMID- 16357805 TI - Increased plasma levels of N-terminal brain natriuretic peptide (NT-proBNP) in type 2 diabetic patients with vascular complications. AB - AIMS: The plasma levels of either brain natriuretic peptide (BNP) or the N terminal fragment of the prohormone (NT-proBNP) have recently gained extreme importance as markers of myocardial dysfunction. Patients with type 2 diabetes are at high risk of developing cardiovascular complications. This study was aimed to assess whether plasma NT-proBNP levels are at similar levels in type 2 diabetics with or without overt cardiovascular diseases. METHODS: We assayed plasma NT-proBNP in 54 type 2 diabetics, 27 of whom had no overt macro- and/or microvascular complications, while the remaining ones had either or both. The same assay was carried out in 38 healthy control subjects age and sex matched as a group with the diabetics. RESULTS: Plasma NT-proBNP was higher in diabetics (median 121 pg/ml, interquartile range 50-240 pg/ml, ) than in those without complications (37 pg/ml, 21-54 pg/ml, P<0.01). Compared with the controls (55 pg/ml, 40-79 pg/ml), only diabetics with vascular complications had significantly increased plasma NT-proBNP levels (P<0.001). In the diabetics, coronary heart disease and nephropathy (defined according to urinary excretion of albumin) were each independently associated with elevated values of plasma NT-proBNP. CONCLUSIONS: In type 2 diabetes mellitus, patients with macro- and/or micro vascular complications exhibit an elevation of plasma NT-proBNP levels compared to corresponding patients with no evidence of vascular disease. The excessive secretion of this peptide is independently associated with coronary artery disease and overt nephropathy. The measurement of circulating NT-proBNP concentration may therefore be useful to screen for the presence of macro- and/or microvascular disease. PMID- 16357806 TI - Correlation between the activities of lipoprotein lipase and paraoxonase in type 2 diabetes mellitus. AB - In type 2 diabetes mellitus the decreased catabolism of triglyceride-rich lipoproteins as a consequence of mainly the decreased lipoprotein lipase activity results in hypertriglyceridaemia and other lipoprotein alterations promoting atherosclerosis. The high-density lipoprotein-associated enzyme, paraoxonase, prevents the oxidation of low-density lipoprotein, which is an antiatherogenic effect. AIM: to examine the relation between the activities of enzymes influencing HDL remodelling- LPL and PON- in type 2 diabetes mellitus. METHODS: 56 newly diagnosed type 2 diabetic patients and 39 healthy controls were involved in the study. The serum PON activity was measured spectrophotometrically using paraoxone as substrate. PON phenotype was determined by the dual substrate method, PON mass was measured by ELISA. The determination of lipoprotein lipase activity was performed using 3H-triolein. RESULTS: We noticed smaller PON activity decrease in our newly diagnosed diabetic subjects compared to the previous studies which investigated the alteration of enzyme activity after a longer duration of diabetes mellitus. The lipoprotein lipase activity showed a positive correlation with PON activity (r=0.43; P<0.02). Interestingly, the PON activity of the homozygous-low activity group did not correlate with the LPL activity, while in the heterozygous and homozygous-high activity groups there was a significantly positive correlation (r=0.51; P<0.05) between PON and LPL activity. CONCLUSION: Besides lipid alterations, the metabolic changes of type 2 diabetes mellitus influence the reduction of the antioxidant capacity of HDL by remodelling HDL and decreasing PON activity via modification of lipoprotein lipase activity, which might contribute to accelerated atherosclerosis. PMID- 16357807 TI - Emerging risk factors and early atherosclerosis indices in subjects with impaired glucose tolerance. AB - AIM: To evaluate the response to an oral lipid overload, inflammatory markers and carotid intima-media thickness in subjects with impaired glucose tolerance. METHODS: 54 subjects, both sexes, 58 y-old average were submitted to 1) Clinical evaluation 2) Glucose tolerance test with 75 g glucose; classified as normal (2 h plasma glucose<140 mg/dl, n=37) or IGT (2 h G 140-200 mg/dl, n=17), 3) 12 h fasting sample (plasma glucose, lipids, C-reactive protein, fibrinogen and HOMA IR calculation); 4 and 6 h after the oral lipid overload (1000 kcal, lipids 65 g) glycemia, fibrinogen and triglycerides were reevaluated. Intima-media thickness was calculated by the average of 6 measurements (3 highest of each carotid) evaluated by ultrasonography (7 MHZ transducer). RESULTS: The IGT group had higher (P<0.001) fasting plasma glucose (89.4 +/- 13 vs 104.4 +/- 8 mg/dl), HOMA IR (1.69 +/- 1.2 vs 2.93 +/- 2.2) and waist (91 +/- 14 vs 101 +/- 9 cm), similar fasting lipids, intima-media thickness (P=0.58) and post-oral lipid overload triglycerides (P=0.74), but higher fibrinogen (284.3 +/- 6 and 305 +/- 10 mg/dl, P=0.05) and C-reactive protein (2.11 +/- 0.33 and 4.19 +/- 0.65 mg/l, P=0.003). C reactive protein was positively correlated with HOMA-IR (r=0.45, P=0.001), fasting plasma glucose (r=0.43, P=0.002) and waist (r=0.45, P=0.0006), but not with postprandial lipids. CONCLUSION: A higher C-reactive protein in IGT, and its positive correlation with insulin resistance indices, but not with postprandial lipaemia, suggests that the clustering of these factors, characteristic of the metabolic syndrome, occurs earlier than postprandial lipid abnormalities. PMID- 16357808 TI - The association between blood glucose value and long-term mortality. AB - This study aims at estimating the association between different fasting blood glucose levels (FBG) and total mortality during a long-term follow-up. In all 2,300 subjects were health examined, out of a stratified sample of 32,185 individuals aged 18-64 years drawn from the population in Stockholm County from the years 1969-70. FBG values were divided into following groups:<3.0, 3.0-4.4, 4.5-5.5, 5.66.0, 6.1-6.6, and > 6.6 mmol/l (corresponding to fasting plasma glucose, FPG,<3.5, 3.5-4.9, 5.0-6.0, 6.1-6.9, 7.0-7.7 and > 7.7 mmol/l), and known diabetes mellitus. All participants were followed up in the National Cause of Death Register up to the end of 1996. Multivariate analysis was performed by Cox regression, with three models, the first age- and sex-adjusted, the second also adjusted for care need category and hypertension, and the third with added BMI-category, with hazard ratios (HR) and 95% confidence interval (CI). Smoking habits were available for around half of the sample. Compared to the FBG showing the lowest mortality, i.e. FBG 5.6-6.0 mmol/l, we found an age- and sex-adjusted excess risk for subjects with known diabetes (HR 7.39, 95% CI 3.78-14.45), with FBG > 6.6 mmol/l (HR 2.30, 95% CI 1.20-4.39), and with FBG<3.0 mmol/l (HR 3.44, 95% CI 1.47-8.06). The excess risk persisted when adjusting for care need, hypertension, BMI, and also for smoking. The cause of the increased mortality risk with low FBG values is unclear, but low FBG value seems to be a risk marker of poor health. PMID- 16357809 TI - Classification of diabetes in young adults: new concepts for an old disease. AB - As clinicians, we are faced to difficult situations in young diabetic patients. The prevalence of type 2 diabetes increases in these patients due to a rising incidence of obesity. We present two clinical observations which both illustrate the insufficiencies of the present classifications. Modern tools are now available for diagnosis such as anti-GAD65 and IA-2 antibodies, genetic tools to investigate for specific mutations, but quantitative means of beta cell mass are lacking. Clinical examination is still accurate to identify type 1 or type 2 diabetes, MODY and mitochondrial diabetes. Weight curve, lesions of acanthosis nigricans, criteria of metabolic syndrome, history of diabetes are critical factors. This problematic has important consequences in our daily practice: the right choice for rapid and good metabolic control. PMID- 16357810 TI - The Ala45Thr polymorphism of NEUROD1 is associated with type 1 diabetes in Brazilian women. AB - BACKGROUND: NEUROD1 encodes a transcription factor expressed in the endocrine pancreas, and involved in beta-cell development, function and mechanisms of apoptosis. In this study, we investigated the association of a frequent polymorphism in exon 2 of NEUROD1 (G > A; Ala45Thr) with Type 1 diabetes in Brazilian subjects. METHODS: A population/association study comprising 246 unrelated Type 1 diabetic and 275 nondiabetic white Brazilian subjects. The Ala45Thr variant was genotyped by a PCR-RFLP method. RESULTS: The frequency of the Thr allele was significantly higher in patients with Type 1 diabetes than in controls (42.3% vs 35.3%, P=0.02). Stratification by gender showed that homozygosity for the Thr allele was associated with Type 1 diabetes in women with odds ratio of 3.66 (95% C.I. 1.43-10.11, P=0.009) as compared to homozygosity for the Ala allele. This effect was not observed in men. CONCLUSIONS: We found a gender-specific association of the Ala45Thr variant of NEUROD1 with Type 1 diabetes in Brazilian women. Our results suggest that gender as well as ethnicity might modulate the association of NEUROD1 with Type 1 diabetes. PMID- 16357811 TI - Factors associated with diabetic patients at high risk for foot ulceration. AB - In order to define risk factors for foot ulcers, associated with the major contributing factors (peripheral sensorimotor neuropathy, peripheral vascular disease, altered foot biomechanics and history of foot ulceration or lower limb amputation), a multivariate analysis was performed in 446 patients free from foot ulceration. Four significant risk factors for foot ulcers have been identified: retinopathy, poor psychosocial status, hyperkeratosis, and diabetes duration. A relation was present between the probability of belonging to the high-risk groups and the number of associated factors. This study points out to the importance of screening especially in case of diabetes of long duration, with microvascular complications, and in socially-deprived people. PMID- 16357812 TI - Pre-diabetes essential action: a European perspective. PMID- 16357813 TI - Prevalence of mitochondrial A3243G mutation in adult type 1 diabetic patients in Catalonia. PMID- 16357814 TI - [Histological and molecular study of fetal human adrenal cortex (12-36 wk)]. AB - Histological and functional characteristics of the fetal human adrenals was studied in 119 normal fetuses aged 12 to 36 weeks development (WD). Immunocytochemical detection of steroidogenesis enzyme (3beta-HSD and P450 c21) and evaluation of cell proliferation using two nuclear markers (Ki-67 and PCNA) were performed in 70 of them. The human fetal adrenal cortex is composed of two morphologically distinct zones: the definitive peripheral zone and the fetal inner zone. From the 12th WD, we observed expression of an adherence protein (NCAM) and two steroidogenesis enzymes (3beta-HSD and P450 c21) in the definitive zone cells, attesting to the capacity of these cells to synthesize mineralocorticoids and/or cortisol. In the fetal zone, only P450 c21 immunoreactivity was detected. From the 14th WD, a transitional zone between the definitive zone and the fetal zone was identified by immunocytochemistry, with expression of 3b-HSD from the 21st WD. Only cells of the definitive zone proliferated from the 12th to 25th WD. The indexes of proliferation of PCNA and Ki-67, 40% and 25% respectively, decreased gradually and were lower than 1% at the 25th WD. PMID- 16357815 TI - Non Hodgkin's lymphoma involving the adrenal glands and the central nervous system (CNS): a particular evolution after chemotherapy. AB - Adrenal lymphoma is extremely rare. The prognostic depends on involvement of other organs (such as the central nervous system) responsible for lower median survival. We report the case of a 51-year-old man with non Hodgkin's Diffuse Large B Cell Lymphoma (DLBCL) involving the central nervous system (CNS) and the adrenal glands simultaneously. The endocrine exploration revealed a partial adrenal insufficiency and ruled out a pheochromocytoma. Computerized tomographic (CT) scan directed needle biopsy of the adrenal gland allowed the diagnostic of non-Hodgkin lymphoma (NHL). CNS biopsies showed similar histopathologic lesions. After aggressive polychemotherapy and methotrexate intrathecal injection, a dissociated therapeutic response was observed with a decrease of the cerebral lesion and an increase of the adrenal mass. This result may be explained by the efficacy of corticosteroid therapy on cerebral edema. The prognosis was poor with tumor infiltration of the leptomeninges and death 16 months after diagnosis. PMID- 16357816 TI - [Hypercalciuria]. AB - The frequency of hypercalciuria is increasing in western countries with an incidence of nephrolithiasis which can reach 13%. Hypercalciuria appears as an alteration of the calcium transport system (kidney, bowel, bone) which is regulated by calcitriol and parathormone. The aim of this review was to screen etiologies of hypercalciuria taking into account recent genetic advances (calcium epithelial channel and calcium sensing receptor). Hypercalciuria may be favored by nutritional causes (diet rich in calcium, sodium, carbohydrates, proteins, poor in phosphates and potassium). It may also be related to an increase in calcium absorption (vitamin D excess, primary hyperparathyroidism, sarcoidosis, lymphoma, estrogens, and certain genetic causes), an increase in osteoresorption (bone metastasis, myeloma, Paget, hyperthyroidism, immobilization, hypercortisolism and corticosteroid therapy), or a decrease of kidney tubular resorption (diuretics, Cacci and Ricci, acromegally, Bartter, familial dominant hypocalcemia, Fanconi, Dent, familial hypomagnesemia-hypercalciuria syndrome, type 1 distal tubular acidosis, pseudohypoaldosteronism, diabetes). If no cause is identified, persistence of hypercalciuria after instituting a correct diet is defined as idiopathic hypercalciuria. Treatment of the cause is essential in secondary hypercalciuria, in addition to diet (low sodium intake, normocalcic diet, hydration), associated with thiazide diuretics and biphosphonates if necessary. PMID- 16357817 TI - [Acromegaly in elderly people]. AB - Acromegaly is associated with increased morbidity and mortality. Cardiovascular complications are the major predictors of mortality. Age and duration of symptoms are the major determinants of acromegalic cardiopathy. Successful control of acromegaly reverse cardiovascular abnormalities. Herein, we review clinical, biological and histopathological findings in elderly people. Treatments and side effects in relation with aging are questioned. PMID- 16357818 TI - [Hypogonadotropic hypogonadism discovered in a patient with cerebellar ataxia]. AB - We present the case of a male patient who presented progressive cerebellar ataxia since childhood who developed partial hypogonadotropic hypogonadism discovered when searching for a cause of low fecundity. The association of these two entities has been described in several rare syndromes: Homes ataxia, Boucher Neuhauser syndrome and Richards-Rundle syndrome. The genetic background of these three syndromic associations defined solely on the basis of clinical manifestations remains to be elucidated, leading to uncertain diagnosis. The present case suggests the syndromic entity could be associated with autosomal recessive spinocerebellar ataxia with hypogonadotropic hypogonadism which includes several genotypic entities. PMID- 16357819 TI - [Pure gonadal dysgenesis XX and XY: observations in fifteen patients]. AB - BACKGROUND: Pure gondal dysgenesis is characterized by impuberism with a female phenotype without genital ambiguity. The aim of the study is to describe the diagnostic and therapeutic patterns as well as the clinical features. PATIENTS AND METHODS: A retrospective study of 15 patients with pure gonadal dysgenesis (15 patients, 46 XX and two 46 XY). Clinical parameters, familial cases, serum gonadotropin levels, pelvic ultrasonography, endoscopic data, karyotype, analysis of SRY (sex determining Y chromosome) and therapeutic control and clinical course were recorded. RESULTS: Average age at diagnosis was 21+/-2.83 years. Primary amenorrhea was the most frequent reason for consultation. A familial case was found in five patients. The association of sensorineural deafness was noted in one patient, suggesting Perrault's syndrome. Serum gonadotropin levels were elevated. Celioscopic evaluation carried out for six patients confirmed the diagnosis. There was one case of uterine and vaginal aplasia association (Mayer Rokytansky-Kuster-Hauser syndrome). In one XY patient, SRY analysis was normal. Prophylactic gonadectomy was performed in both XY patients. Substitution therapy was initiated in 11 patients. Follow-up in 6 patients revealed development of secondary sexual characters. DISCUSSION: The clinical, biological and histological features of our patients presenting pure gonadal dysgenesis XX were in agreement with earlier reports in the literature. Familial cases suggest possible autosomal transmission. The lack of a mutation in XY patients suggests a post-transcription anomaly. Complete or parital dysgenesis can be identified by histological analysis of the gondads. CONCLUSION: Study of sex determining genes should provide new perspectives for earlier diagnosis and treatment of pure gondadal dysgenesis. PMID- 16357821 TI - West Nile virus activity--United States, January 1-December 1, 2005. AB - West Nile virus (WNV) is the leading cause of arboviral encephalitis in the United States. Originally discovered in Africa in 1937, WNV was first detected in the western hemisphere in 1999 in New York City. Since then it has caused seasonal epidemics of febrile illness and severe neurologic disease. During January 1-December 1, 2005, a total of 2,744 cases of WNV disease in humans were reported in the United States, an increase from 2,359 during the same period in 2004. A total of 1,165 cases were WNV neuroinvasive disease (WNND). WNV infections in humans, birds, mosquitoes, and nonhuman mammals are reported to CDC through ArboNET, an Internet-based arbovirus surveillance system managed by state health departments and CDC. During 2005, WNV transmission to humans or animals expanded into 21 counties that had not previously reported transmission and recurred in 1,196 counties where transmission had been reported in previous years. This report summarizes provisional WNV surveillance data through December 1, 2005, and highlights the need for ongoing surveillance, mosquito control, promotion of personal protection from mosquito bites, and research into additional prevention strategies. PMID- 16357822 TI - Update: influenza activity--United States, October 2-December 3, 2005. AB - During October 2-December 3, 2005, low level influenza activity was reported in the United States. This report summarizes U.S. influenza activity since the beginning of the 2005-06 influenza surveillance season and updates the previous summary. PMID- 16357823 TI - Guidelines for the investigation of contacts of persons with infectious tuberculosis. Recommendations from the National Tuberculosis Controllers Association and CDC. AB - In 1976, the American Thoracic Society (ATS) published brief guidelines for the investigation, diagnostic evaluation, and medical treatment of TB contacts. Although investigation of contacts and treatment of infected contacts is an important component of the U.S. strategy for TB elimination, second in priority to treatment of persons with TB disease, national guidelines have not been updated since 1976. This statement, the first issued jointly by the National Tuberculosis Controllers Association and CDC, was drafted by a working group consisting of members from both organizations on the basis of a review of relevant epidemiologic and other scientific studies and established practices in conducting contact investigations. This statement provides expanded guidelines concerning investigation of TB exposure and transmission and prevention of future cases of TB through contact investigations. In addition to the topics discussed previously, these expanded guidelines also discuss multiple related topics (e.g., data management, confidentiality and consent, and human resources). These guidelines are intended for use by public health officials but also are relevant to others who contribute to TB control efforts. Although the recommendations pertain to the United States, they might be adaptable for use in other countries that adhere to guidelines issued by the World Health Organization, the International Union against Tuberculosis and Lung Disease, and national TB control programs. PMID- 16357824 TI - Guidelines for using the QuantiFERON-TB Gold test for detecting Mycobacterium tuberculosis infection, United States. AB - On May 2, 2005, a new in vitro test, QuantiFERON-TB Gold (QFT-G, Cellestis Limited, Carnegie, Victoria, Australia), received final approval from the U.S. Food and Drug Administration as an aid for diagnosing Mycobacterium tuberculosis infection. This test detects the release of interferon-gamma (IFN-g) in fresh heparinized whole blood from sensitized persons when it is incubated with mixtures of synthetic peptides representing two proteins present in M. tuberculosis: early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10). These antigens impart greater specificity than is possible with tests using purified protein derivative as the tuberculosis (TB) antigen. In direct comparisons, the sensitivity of QFT-G was statistically similar to that of the tuberculin skin test (TST) for detecting infection in persons with untreated culture-confirmed tuberculosis (TB). The performance of QFT-G in certain populations targeted by TB control programs in the United States for finding latent TB infection is under study. Its ability to predict who eventually will have TB disease has not been determined, and years of observational study of substantial populations would be needed to acquire this information. In July 2005, CDC convened a meeting of consultants and researchers with expertise in the field to review scientific evidence and clinical experience with QFT-G. On the basis of this review and discussion, CDC recommends that QFT-G may be used in all circumstances in which the TST is currently used, including contact investigations, evaluation of recent immigrants, and sequential-testing surveillance programs for infection control (e.g., those for health-care workers). This report provides specific cautions for interpreting negative QFT-G results in persons from selected populations. This report is aimed at public health officials, health-care providers, and laboratory workers with responsibility for TB control activities in the United States. PMID- 16357825 TI - The regulatory subunit of PDE6 interacts with PACSIN in photoreceptors. AB - PURPOSE: In photoreceptors, phosphodiesterase 6 (PDE6) is regulated in response to light, due to the shuttling of a regulatory subunit (PDE6gamma) between the catalytic subunits of PDE6 and the activated form of transducin. We showed previously that PDE6gamma is able to interact with the Src-homology type 3 (SH3) domain of formin-binding protein 17 (FBP17), a protein involved in membrane receptor endocytosis. FBP17 was not detected in rat retina. Therefore, we looked for other SH3 domain-containing proteins that might interact with PDE6gamma in rat photoreceptors. METHODS: Several SH3 domains highly homologous to this domain of FBP17 were found by structural alignment. Yeast two-hybrid system and GST pull downs were used to test interaction of PDE6gamma with these putative partners. Expression patterns in rat retina of the SH3 containing candidates were also determined by immunohistochemistry and western blotting. GST pull-downs and co immunopreciptations were then used to test in vivo interaction with PDE6gamma in rat retina extracts. Colocalization and light translocation of PDE6gamma and one of its partner were studied by confocal microscopy. RESULTS: PDE6gamma interacts in vitro with a number of SH3 domains. These interactions involve a polyprolin motif located between amino acids 20 and 28 of PDE6gamma. Several of the putative partners of PDE6gamma are expressed in photoreceptor cells and might therefore interact in vivo with PDE6gamma. Our results show that only PACSIN, a protein implicated in endocytosis, was found to interact with PDE6gamma in rat retina extracts. The colocalization of the two proteins occurs in photoreceptor inner segments and synapses and is greatly enhanced upon illumination of the retina. CONCLUSIONS: PDE6gamma function is mostly documented in the regulation of phototransduction. Our results provide evidence that in vitro PDE6gamma has a broad pattern of SH3 containing partners expressed in photoreceptors. PDE6gamma interaction with PACSIN points to a possible role of PDE6gamma in endocytosis. Further studies will be needed to understand the exact role of PDE6gamma-PACSIN interactions in photoreceptors. The description of this new function of PDE6gamma might help to understand the molecular mechanism of the severe retinal degeneration observed in PDE6gamma knock-out mice. PMID- 16357826 TI - Use of transduction proteins to target trabecular meshwork cells: outflow modulation by profilin I. AB - PURPOSE: Fusion proteins containing a protein transduction domain (PTD4) are able to cross biological membranes. We tested the applicability of the protein transduction method for study of the aqueous humor trabecular outflow pathway by targeting the actin cytoskeleton, which is known to be involved in outflow facility regulation. METHODS: Expression vectors useful for generating fusion proteins with the PTD4 domain and the actin-binding protein Profilin I were constructed. The transductional and functional properties of these proteins were tested in bovine trabecular meshwork cells in culture. The effects of PTD4 Profilin I on outflow facility were evaluated in perfused bovine anterior segments. PTD4-beta-galactosidase was used to visually check correct delivery of fusion proteins to trabecular meshwork cells. RESULTS: The fusion proteins generated were characterized by western blot. Immunocytochemistry experiments showed intracellular staining for PTD4-Profilin I in trabecular meshwork cells in culture. The fusion protein was found in the cytoplasm associated with actin filaments and in the leading edge of the cellular membrane. In contrast, control Profilin I, without the PTD4 domain, was unable to cross the cell membrane. In perfused anterior segments, 2 microM PTD4-Profilin I increased trabecular outflow facility in a reversible manner, while Profilin I had no significant effect. Anterior segments perfused with PTD4-beta-galactosidase showed positive staining in the trabecular meshwork tissue. CONCLUSIONS: Protein transduction technology is a valuable tool for targeting trabecular meshwork tissue, not only for performing physiological studies, but also as a potential drug-delivery method. Profilin I action on the actin cytoskeleton further reinforces the importance of this structure in outflow facility regulation. PMID- 16357827 TI - Conserved structure and spatiotemporal function of the compact rhodopsin kinase (GRK1) enhancer/promoter. AB - PURPOSE: To demonstrate that the crucial elements responsible for the spatial and temporal expression patterns of rhodopsin kinase (Rk) are contained within a narrow conserved segment immediately flanking the Rk transcription start sites. METHODS: Sequences upstream of the mouse Rk gene were compared to the human sequence to identify areas of conservation. Transgenic mice carrying a segment of the conserved human DNA sequence linked upstream of the green fluorescent protein (GFP) gene were examined by fluorescence microscopy and RT-PCR to localize GFP expression in retina and pineal gland. Rk and GFP temporal expression patterns were further compared by immunostaining and real-time RT-PCR in transgenic eyes during development. RESULTS: Comparison of the mouse and human 5' flanking sequence revealed only a small island of conserved sequence upstream of the respective Rk start sites. Uniform GFP expression was supported by a 0.2 kb fragment of the conserved human sequence in the transgenic mouse rods, cones, and pinealocytes. Developmental studies revealed an exponential rise in Rk and GFP transcripts in the first ten day postnatal period followed by a plateau later extending to adulthood. Rk and GFP proteins were first detected after postnatal day 10 and rose in parallel afterwards, overlapping in time with the maturation of photoreceptor outer segments and eye opening. CONCLUSIONS: The conserved short enhancer/promoter immediately upstream of the Rk gene contains the key elements required for appropriate response to spatial and temporal cues during photoreceptor cell differentiation and fate determination. The above studies narrow the core sequences that govern gene expression in photoreceptors in vivo. PMID- 16357828 TI - Spatial and temporal expression patterns of the choroideremia gene in the mouse retina. AB - PURPOSE: Choroideremia (CHM), an X-linked retinal disease, is caused by mutations affecting the CHM gene. This gene encodes REP-1, which functions in the covalent modifications of proteins involved in vesicle trafficking. The disease affects several cell types in the retina, but it is not known which cell types contribute directly or indirectly to disease progression. A study of the expression patterns of Chm and the related gene Chml in the mouse retina was undertaken in order to address this issue. METHODS: The expression patterns of Chm and Chml were determined by in situ hybridization. The localization of the Chm protein product, Rep-1, was determined spatially and temporally in the mouse retina by immunohistochemistry. RESULTS: Chm and Chml mRNA were found in every major layer of the retina in adult mice. During development, Rep-1 protein localization changes from a fairly diffuse pattern during embryogenesis to a more specific pattern at the time of retinal differentiation. In adulthood, Rep-1 localizes to distinct cellular compartments in multiple retinal cell types. CONCLUSIONS: Chm and Chml have the same broad expression profile in the mouse retina. In particular, the Chm transcript and corresponding protein are found in cell types other than those thought to be primarily affected in the human disease. These results have important implications for approaches with which to develop a relevant mouse model of choroideremia and for therapeutic strategies for this disease. PMID- 16357829 TI - Cryptic chromosomal aberrations waiting to be discovered. PMID- 16357830 TI - Clonotypic analysis of acute lymphoblastic leukemia with a double TEL-AML1 fusion at onset and relapse. PMID- 16357831 TI - A novel and cytogenetically cryptic t(7;21)(p22;q22) in acute myeloid leukemia results in fusion of RUNX1 with the ubiquitin-specific protease gene USP42. AB - Although many of the chromosomal abnormalities in hematologic malignancies are identifiable cytogenetically, some are only detectable using molecular methods. We describe a novel cryptic t(7;21)(p22;q22) in acute myeloid leukemia (AML). FISH, 3'RACE, and RT-PCR revealed a fusion involving RUNX1 and the ubiquitin specific protease (USP) gene USP42. The genomic breakpoint was in intron 7 of RUNX1 and intron 1 of USP42. The reciprocal chimera was not detected - neither on the transcriptional nor on the genomic level - and FISH showed that the 5' part of USP42 was deleted. USP42 maps to a 7p22 region characterized by segmental duplications. Notably, 17 kb duplicons are present 1 Mb proximal to USP42 and 3 Mb proximal to RUNX1; these may be important in the genesis of t(7;21). This is the second cryptic RUNX1 translocation in hematologic malignancies and the first in AML. The USPs have not previously been reported to be rearranged in leukemias. The cellular context in which USP42 is active is unknown, but we here show that it is expressed in normal bone marrow, in primary AMLs, and in cancer cell lines. Its involvement in the t(7;21) suggests that deregulation of ubiquitin-associated pathways may be pathogenetically important in AML. PMID- 16357832 TI - Target for cancer therapy: proliferating cells or stem cells. AB - Tumor stem cells are quiescent and, therefore, resistant to therapy, yet harbor the capacity to replenish a tumor after therapy. Therefore, it is tempting to explain all therapeutic failures by the persistence of tumor stem cells. Yet, this explanation is relevant only to initial stages of stem-cell-dependent tumors (such as chronic myeloid leukemia) that, actually, are well controlled by therapy. In advanced cancers that poorly respond to therapy, quiescent tumor stem cells play a negligible role. Instead, proliferating cells determine disease progression, prognosis, therapeutic failures, and resistance to therapy. And therapy fails not because it eliminates only proliferating tumor cells, but because it does not eliminate them. With noticeable exceptions, it is the proliferating cell that should be targeted, whereas resting cancer cells including stem and dormant cells need to be targeted only when they 'wake up'. Finally, I discuss a strategy of selectively killing dominant proliferating clones, including proliferating stem-like and drug-resistant cancer cells, while sparing normal cells. PMID- 16357833 TI - Silencing of the tumor suppressor gene FHIT is highly characteristic for MLL gene rearranged infant acute lymphoblastic leukemia. AB - MLL rearranged acute lymphoblastic leukemia (MLL) is an aggressive type of acute lymphoblastic leukemia (ALL), diagnosed predominantly in infants (<1 years of age). Since current chemotherapy fails in >50% of patients with MLL, new therapeutic strategies are desperately needed. For this, understanding the biological features characterizing MLL is necessary. Analysis of gene expression profiles revealed that the expression of the tumor suppressor gene FHIT is reduced in children with MLL rearranged ALL as compared to ALL patients carrying germ line MLL. This finding was confirmed by quantitative real-time PCR. In 100% of the infant MLL cases tested, methylation of the FHIT 5'CpG region was observed, resulting in strongly reduced mRNA and protein expression. In contrast, FHIT methylation in infant and non-infant ALL patients carrying germ line MLL was found in only approximately 60% (P< or =0.004). FHIT expression was restored upon exposing leukemic cells to the demethylating agent decitabine, which induced apoptosis. Likewise and more specifically, leukemic cell death was induced by transfecting MLL rearranged leukemic cells with expression vectors encoding wild type FHIT, confirming tumor suppressor activity of this gene. These observations imply that suppression of FHIT may be required for the development of MLL, and provide new insights into leukemogenesis and therapeutic possibilities for MLL. PMID- 16357834 TI - The nuclear oncoprotein TLX1/HOX11 associates with pericentromeric satellite 2 DNA in leukemic T-cells. AB - TLX1/HOX11, a DNA-binding homeodomain protein, was originally identified by virtue of its aberrant expression in T-cell leukemia and subsequently found to be crucial for normal spleen development. The precise mechanism of TLX1 function remains poorly understood, although it is known that it can act as both a transcriptional activator and repressor and can downregulate the Aldh1a1 gene in embryonic mouse spleen. Using a whole-genome PCR approach, we show here that TLX1 protein directly interacts with pericentromeric human satellite 2 DNA sequences. Such DNA is known to localize to heterochromatin, which among other roles has been implicated in gene silencing. The interaction was confirmed in vitro and in vivo by gel retardation and chromatin immunoprecipitation assays involving satellite 2 DNA, which contained sequences resembling TLX1 binding sites. Using immunofluorescence microscopy, TLX1 demonstrated a punctate pattern of staining in the nuclei of leukemic T-cells (ALL-SIL). Double labelling indicated that TLX1 colocalized with the centromeric protein CENP-B, demonstrating that the TLX1 foci corresponded to clusters of centromeric DNA. The novel interaction of TLX1 with constitutive heterochromatin adds an additional level of complexity to the intracellular functions of this transcriptional regulator and may have relevance to its roles in transcriptional repression and T-cell immortalization. PMID- 16357835 TI - Platelet transfusion can mimic somatic mtDNA mutations. PMID- 16357836 TI - Bone marrow angiogenesis in multiple myeloma. AB - Angiogenesis is a constant hallmark of multiple myeloma (MM) progression and has prognostic potential. It is induced by plasma cells via angiogenic factors with the transition from monoclonal gammopathy of undetermined significance (MGUS) to MM, and probably with loss of angiostatic activity on the part of MGUS. The pathophysiology of MM-induced angiogenesis is complex and involves both direct production of angiogenic cytokines by plasma cells and their induction within the microenvironment. The latter are secreted by stromal cells, endothelial cells (EC) and osteoclasts, and promote plasma cell growth, survival and migration, as well as paracrine cytokine secretion and angiogenesis in the bone marrow milieu. Angiogenesis is also supported by inflammatory cells following their recruitment and activation by plasma cells. Finally, circulating EC and endothelial precursor cells (EPC) contribute to the neovascularization, and the presence of EPC suggests that vasculogenesis (new vessel formation from EPC) may also contribute to the full MM vascular tree. PMID- 16357837 TI - Functional MR imaging of tumor angiogenesis predicts outcome of patients with acute myeloid leukemia. PMID- 16357838 TI - Autologous stem cell transplantation in adults with acute lymphoblastic leukemia in first complete remission: analysis of the LALA-85, -87 and -94 trials. AB - To evaluate the results of autologous stem cell transplantation (ASCT) in a large population of adults with acute lymphoblastic leukemia (ALL) in first complete remission (CR), we performed an individual data-based overview of the last three trials from the LALA group. Overall, 349 patients with ALL prospectively randomized in the consecutive LALA-85, -87, and -94 trials to receive either ASCT or chemotherapy as post-CR treatment were analyzed. Eligibility criteria were 15 50-year-old patients without sibling donors in both LALA-85/87 trials and 15-55 year-old patients with high-risk ALL and no sibling donors in the LALA-94 trial. Intent-to-treat analysis, which compared 175 patients from the ASCT arm to 174 patients from the chemotherapy arm, showed that ASCT was associated with a lower cumulative incidence of relapse (66 vs 78% at 10 years; P=0.05), without significant gain in disease-free or overall survival. Despite a possible lack of statistical power, a nested case-control analysis performed in 85 patient pairs adjusted for time to transplant and prognostic covariates confirmed these intent to-treat results in patients actually transplanted. Of interest, the reduced relapse risk after ASCT translated in better disease-free survival in the 300 rapid responders who reached CR after the first induction course. PMID- 16357839 TI - Human cytotoxic T lymphocytes specific for a single minor histocompatibility antigen HA-1 are effective against human lymphoblastic leukaemia in NOD/scid mice. PMID- 16357840 TI - Diagnosis of occult tuberculosis in hematological malignancy by enumeration of antigen-specific T cells. PMID- 16357841 TI - Pilot study of combination transcriptional modulation therapy with sodium phenylbutyrate and 5-azacytidine in patients with acute myeloid leukemia or myelodysplastic syndrome. AB - Epigenetic mechanisms underlying tumorigenesis have recently received much attention as potential therapeutic targets of human cancer. We designed a pilot study to target DNA methylation and histone deacetylation through the sequential administration of 5-azacytidine followed by sodium phenylbutyrate (PB) in patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). Ten evaluable patients (eight AML, two MDS) were treated with seven consecutive daily subcutaneous injections of 5-azacytidine at 75 mg/m2 followed by 5 days of sodium PB given intravenously at a dose of 200 mg/kg. Five patients (50%) were able to achieve a beneficial clinical response (partial remission or stable disease). One patient with MDS proceeded to allogeneic stem cell transplantation and is alive without evidence of disease 39 months later. The combination regimen was well tolerated with common toxicities of injection site skin reaction (90% of the patients) from 5-azacytidine, and somnolence/fatigue from the sodium PB infusion (80% of the patients). Correlative laboratory studies demonstrated the consistent reacetylation of histone H4, although no relationship with the clinical response could be demonstrated. Results from this pilot study demonstrate that a combination approach targeting different mechanisms of transcriptional modulation is clinically feasible with acceptable toxicity and measurable biologic and clinical outcomes. PMID- 16357842 TI - Keratin expression in schwannoma; a study of 115 retroperitoneal and 22 peripheral schwannomas. AB - Schwannomas have been variably observed to be glial fibrillary acid protein (GFAP) and occasionally keratin positive, with antibodies reacting with multiple keratins (pankeratins, keratin cocktail (CK), but specific keratin polypeptides (K) have not been examined for in schwannoma. Since we observed CK positivity in retroperitoneal schwannomas, we wanted to study a large group of retroperitoneal and peripheral schwannomas with GFAP, CK and Ks to explore the frequency and biologic background of this finding. We immunohistochemically evaluated a large number of retroperitoneal (n=115) and peripheral schwannomas (n=22) for GFAP, 16 individual K and AE1/AE3 keratin cocktail. The great majority (104/115, 90%) of retroperitoneal schwannomas were positive for GFAP, and 72/104 (69%) cases were positive for AE1/AE3, often extensively. Both markers highlighted the cellular Antoni A areas, particularly adjacent to the capsule, myxoid or degenerative areas, and perivascularly. Most cases 87/104 (84%) stained for both AE1/AE3 and GFAP at least focally. No tumors stained for keratins that were GFAP negative. None of the immunostains for individual K showed positivity comparable to that obtained with AE1/AE3 CK. However, 62% were focally positive for high molecular weight K1 and 8/61 (13%) for K7. None of the retroperitoneal schwannomas were positive for other keratins including K2, 4, 5, 8, 9, 10 and K14-20. Peripheral schwannomas showed GFAP-positivity in only three of 22 cases (14%), and all were negative for keratins, both cocktail and individual K. We conclude that crossreactivity of AE1/AE3 with other intermediate filament proteins, such as GFAP, as previously observed in brain and glioma tissue, probably accounts for the extensive keratin-positivity seen in some retroperitoneal schwannomas. However, focal expression of K1 and K7 cannot be ruled out. Keratin-positive schwannomas should not be confused with other keratin-positive tumors, such as sarcomatoid carcinoma, mesothelioma, and synovial sarcoma. PMID- 16357843 TI - Molecular and immunohistochemical analyses of the focal form of congenital hyperinsulinism. AB - Congenital hyperinsulinism is a rare pancreatic endocrine cell disorder that has been categorized histologically into diffuse and focal forms. In focal hyperinsulinism, the pancreas contains a focus of endocrine cell adenomatous hyperplasia, and the patients have been reported to possess paternally inherited mutations of the ABCC8 and KCNJ11 genes, which encode subunits of an ATP sensitive potassium channel (K(ATP)). In addition, the hyperplastic endocrine cells show loss of maternal 11p15, where imprinted genes such as p57(kip2) reside. In order to evaluate whether all cases of focal hyperinsulinism are caused by this mechanism, 56 pancreatectomy specimens with focal hyperinsulinism were tested for the loss of maternal allele by two methods: immunohistochemistry for p57(kip2) (n=56) and microsatellite marker analysis (n=27). Additionally, 49 patients were analyzed for K(ATP) mutations. Out of 56 focal lesions, 48 demonstrated clear loss of p57(kip2) expression by immunohistochemistry. The other eight lesions similarly showed no nuclear labeling, but the available tissue was not ideal for definitive interpretation. Five of these eight patients had paternal K(ATP) mutations, of which four demonstrated loss of maternal 11p15 within the lesion by microsatellite marker analysis. All of the other three without a paternal K(ATP) mutation showed loss of maternal 11p15. K(ATP) mutation analysis identified 32/49 cases with paternal mutations. There were seven patients with nonmaternal mutations whose paternal DNA material was not available, and one patient with a mutation that was not present in either parent's DNA. These eight patients showed either loss of p57(kip2) expression or loss of maternal 11p15 region by microsatellite marker analysis, as did the remaining nine patients with no identifiable K(ATP) coding region mutations. The combined results from the immunohistochemical and molecular methods indicate that maternal 11p15 loss together with paternal K(ATP) mutation is the predominant causative mechanism of focal hyperinsulinism. PMID- 16357844 TI - Benign metastasizing leiomyoma: clonality, telomere length and clinicopathologic analysis. AB - Benign metastasizing leiomyoma is a rare condition affecting women with a history of uterine leiomyomata and is characterized by multiple histologically benign pulmonary smooth muscle tumors. Speculations on its pathogenesis include a benign uterine leiomyoma colonizing the lung, a metastatic low-grade uterine leiomyosarcoma, and primary pulmonary leiomyomatosis. To elucidate its pathogenesis, we analyzed the clinical, pathological and immunohistochemical features, clonality, and telomere length of multiple lung and uterine tumors in three patients with benign metastasizing leiomyoma. In all cases, pulmonary tumors had benign histology and immunohistochemical profiles (estrogen receptor positive, progesterone receptor positive, and very low proliferative index) identical to uterine leiomyoma. In eight tumors from three patients, clonality was assessed by analyzing the variable length of the polymorphic CAG repeat sequence within the human androgen receptor gene. In the two informative patients pulmonary and uterine tumors showed identical patterns of androgen receptor allelic inactivation, indicating that they were clonal. The telomere length measured by fluorescence in situ hybridization in pulmonary leiomyomas of all three patients were either long or very long and were identical to the uterine counterparts, indicating significant telomere shortening is not a crucial step for developing metastases. Our evidence supports the notion that benign metastasizing leiomyoma is clonally derived from benign-appearing uterine leiomyomas. PMID- 16357845 TI - Methylation of p16(INK4A) and p57(KIP2) are involved in the development and progression of gastric MALT lymphomas. AB - p16(INK4A) and p57(KIP2) are inhibitors of cyclin-dependent kinases and their inactivation by methylation has been reported as a major tumorigenic mechanism in tumors. To examine whether methylation of p16(INK4A) and p57(KIP2) is involved in the development and progression of gastric MALT lymphomas, 24 gastric low-grade lymphomas of MALT, 11 diffuse large B-cell lymphomas, and 10 each case of gastric lymphoid follicles with and without Helicobacter pylori infection were studied. H. pylori infection was positive in 85.7% of the gastric lymphomas. In the gastric lymphoid follicles positive for H. pylori, methylation of p16(INK4A) was detected in 10% of cases, while methylation of p57(KIP2) was not detected. In low grade MALT lymphomas, p16(INK4A) and p57(KIP2) were methylated in 41.7 and 29.2% of the cases, respectively. In diffuse large B-cell lymphomas, methylation of p16(INK4A) and p57(KIP2) was found in 72.7 and 36.4% of the cases, respectively. All but one case with p16(INK4A) and p57(KIP2) methylation was H. pylori positive and most of them were stage I. Our results indicate that methylation of p16(INK4A) followed by p57(KIP2) methylation involves during the tumorigenesis of gastric MALT lymphomas associated with H. pylori infection. As methylation of these two genes was more frequent in the higher grade (P<0.05), it may contribute to the malignant progression of gastric MALT lymphomas. PMID- 16357846 TI - Scientific blues. PMID- 16357847 TI - Vestigial no more. PMID- 16357848 TI - The 'chop-and-change' of MHC class I assembly. PMID- 16357849 TI - Orchestrated leak provokes a thymus reassessment. PMID- 16357850 TI - Lymph node choreography: Treg cells join the dance. PMID- 16357851 TI - Toll-erating self DNA. PMID- 16357852 TI - A TRAFfic cop for host defense. PMID- 16357854 TI - HIV vaccine design: insights from live attenuated SIV vaccines. AB - The International AIDS Vaccine Initiative has established a consortium to elucidate mechanisms of protection conferred by live attenuated simian immunodeficiency virus vaccines in monkeys. Here, the strategies defining key components of the protective immune response elicited by these vaccines are discussed. PMID- 16357855 TI - Lymphocyte calcium signaling from membrane to nucleus. AB - Ca(2+) signals control a variety of lymphocyte responses, ranging from short-term cytoskeletal modifications to long-term changes in gene expression. The identification of molecules and channels that modulate Ca(2+) entry into T and B lymphocytes has both provided details of the molecular events leading to immune responses and raised controversy. Here we review studies of the pathways that allow Ca(2+) entry, the function of Ca(2+) in the regulation of cell polarity and motility and the principles by which Ca(2+)-dependent transcription regulates lymphocyte function. PMID- 16357857 TI - Meningococcal genome dynamics. AB - Neisseria meningitidis (the meningococcus) is an important commensal, pathogen and model organism that faces up to the environment in its exclusive human host with a small but hyperdynamic genome. Compared with Escherichia coli, several DNA repair genes are absent in N. meningitidis, whereas the gene products of others interact differently. Instead of responding to external stimuli, the meningococcus spontaneously produces a plethora of genetic variants. The frequent genomic alterations and polymorphisms have profound consequences for the interaction of this microorganism with its host, impacting structural and antigenic changes in crucial surface components that are relevant for adherence and invasion as well as antibiotic resistance and vaccine development. PMID- 16357858 TI - Hendra and Nipah viruses: different and dangerous. AB - Hendra virus and Nipah virus are highly pathogenic paramyxoviruses that have recently emerged from flying foxes to cause serious disease outbreaks in humans and livestock in Australia, Malaysia, Singapore and Bangladesh. Their unique genetic constitution, high virulence and wide host range set them apart from other paramyxoviruses. These features led to their classification into the new genus Henipavirus within the family Paramyxoviridae and to their designation as Biosafety Level 4 pathogens. This review provides an overview of henipaviruses and the types of infection they cause, and describes how studies on the structure and function of henipavirus proteins expressed from cloned genes have provided insights into the unique biological properties of these emerging human pathogens. PMID- 16357859 TI - Antimicrobial-resistant bacteria in the community setting. AB - Over the past decade, antimicrobial resistance has emerged as a major public health crisis. Common bacterial pathogens in the community such as Streptococcus pneumoniae have become progressively more resistant to traditional antibiotics. Salmonella strains are beginning to show resistance to crucial fluoroquinolone drugs. Community outbreaks caused by a resistant form of Staphylococcus aureus, known as community-associated meticillin (formerly methicillin)-resistant Staphylococcus aureus, have caused serious morbidity and even deaths in previously healthy children and adults. To decrease the spread of such antimicrobial-resistant pathogens in the community, a greater understanding of their means of emergence and survival is needed. PMID- 16357860 TI - Modelling strategies for the industrial exploitation of lactic acid bacteria. AB - Lactic acid bacteria (LAB) have a long tradition of use in the food industry, and the number and diversity of their applications has increased considerably over the years. Traditionally, process optimization for these applications involved both strain selection and trial and error. More recently, metabolic engineering has emerged as a discipline that focuses on the rational improvement of industrially useful strains. In the post-genomic era, metabolic engineering increasingly benefits from systems biology, an approach that combines mathematical modelling techniques with functional-genomics data to build models for biological interpretation and--ultimately--prediction. In this review, the industrial applications of LAB are mapped onto available global, genome-scale metabolic modelling techniques to evaluate the extent to which functional genomics and systems biology can live up to their industrial promise. PMID- 16357861 TI - Advances in understanding bacterial outer-membrane biogenesis. AB - The outer membrane of gram-negative bacteria such as Escherichia coli serves as a protective barrier that controls the influx and efflux of solutes. This allows the bacteria to inhabit several different, and often hostile, environments. The assembly of the E. coli outer membrane has been difficult to study using traditional genetic and biochemical methods, and how all its components reach the outer membrane after being synthesized in the cytoplasm and cytoplasmic membrane, how they are assembled in an environment that is devoid of an obvious energy source, and how assembly proceeds without disrupting the integrity of this essential cellular structure are all fundamental questions that remain unanswered. Here, we review the new approaches that have led to the recent discovery of components of the machinery involved in the biogenesis of this distinctive cellular organelle. PMID- 16357862 TI - Virus membrane-fusion proteins: more than one way to make a hairpin. AB - Structure-function studies have defined two classes of viral membrane-fusion proteins that have radically different architectures but adopt a similar overall 'hairpin' conformation to induce fusion of the viral and cellular membranes and therefore initiate infection. In both classes, the hairpin conformation is achieved after a conformational change is triggered by interaction with the target cell. This review will focus in particular on the properties of the more recently described class II proteins. PMID- 16357863 TI - The poetry of science. AB - During the past year, I have taken part in an experiment in which I have been the experimental animal. I was awarded a fellowship that gave me the opportunity to take a sabbatical from science and spend a year in the School of Literature and Creative Writing at the University of East Anglia. The School of Literature and Creative Writing is internationally renowned, and past students include household names such as Ian McEwan, Kazuo Ishiguro, Trezza Azzopardi and Tracy Chevalier. I have a keen interest in creative writing and had written a few short stories. This was a formidable but thrilling challenge. During my adventure, I realized that, in many ways, I was coming home. PMID- 16357866 TI - Lipopolysaccharide signaling in endothelial cells. AB - Sepsis is the systemic immune response to severe bacterial infection. The innate immune recognition of bacterial and viral products is mediated by a family of transmembrane receptors known as Toll-like receptors (TLRs). In endothelial cells, exposure to lipopolysaccharide (LPS), a major cell wall constituent of Gram-negative bacteria, results in endothelial activation through a receptor complex consisting of TLR4, CD14 and MD2. Recruitment of the adaptor protein myeloid differentiation factor (MyD88) initiates an MyD88-dependent pathway that culminates in the early activation of nuclear factor-kappaB (NF-kappaB) and the mitogen-activated protein kinases. In parallel, a MyD88-independent pathway results in a late-phase activation of NF-kappaB. The outcome is the production of various proinflammatory mediators and ultimately cellular injury, leading to the various vascular sequelae of sepsis. This review will focus on the signaling pathways initiated by LPS binding to the TLR4 receptor in endothelial cells and the coordinated regulation of this pathway. PMID- 16357867 TI - Immunization in familial amyloidotic polyneuropathy: counteracting deposition by immunization with a Y78F TTR mutant. AB - The mechanism of amyloid formation in familial amyloidotic polyneuropathy (FAP), a hereditary disorder associated with mutant transthyretin (TTR), is still unknown. It is generally believed that altered conformations exposing cryptic regions are intermediary steps in this mechanism. A TTR mutant--Y78F (transthyretin mutant with phenylalanine replacing tyrosine at position 78)- designed to destabilize the native structure has been shown to expose a cryptic epitope recognized by a monoclonal antibody that reacts only with highly amyloidogenic mutants presenting the amyloid fold or with amyloid fibrils. To test whether TTR deposition in FAP can be counteracted by antibodies for cryptic epitopes, we immunized with TTR Y78F, transgenic mice carrying the most common FAP-associated TTR mutant--V30M (transthyretin mutant with methionine replacing valine at position 30)--at selected ages that present normally with either nonfibrillar or TTR amyloid deposition. Compared to age-matched control nonimmunized mice, Y78F-immunized mice had a significant reduction in TTR deposition usually found in this strain, in particular in stomach and intestine; by contrast, animals immunized with V30M did not show differences in deposition in comparison with nonimmunized mice. Immunohistochemical analyses of tissues revealed that immunization with Y78F lead to infiltration by lymphocytes and macrophages at common deposition sites, but not in tissues such as liver, choroid plexus, and Langerhans islets, in which TTR is produced. These results suggest that Y78F induced production of an antibody that reacts specifically with deposits and leads to an immune response effective in removing/preventing TTR deposition. Therefore, TTR immunization with selected TTR mutants has potential application in immune therapy for FAP. PMID- 16357868 TI - Heme oxygenase-1 mediates the protective effects of rapamycin in monocrotaline induced pulmonary hypertension. AB - Rapamycin inhibits the development and progression of vascular disease. We previously showed that rapamycin induces the cytoprotective protein, heme oxygenase-1 (HO-1), and more importantly, chemically inhibiting HO-1 blocked the antiproliferative actions of rapamycin. In this study, we evaluated whether HO-1 is required for the vascular protective effects of rapamycin in vivo using a rat monocrotaline-induced pulmonary hypertension model. Rats were exposed to monocrotaline with or without rapamycin and HO activity was altered using the chemical inhibitor, tin protoporphyrin or the inducer, cobalt protoporphyrin. We also evaluated possible mechanisms of rapamycin-dependent induction of HO-1, and how HO-1 mediates growth factor-dependent antiproliferative actions of rapamycin. Proliferation and cell cycle progression were examined in smooth muscle cells derived from both wild-type and HO-1 knockout (HO-1-/-) mice in response to growth factors and rapamycin. Similar to our previous findings in vitro, rapamycin induced HO-1 in rat lung. Rapamycin also inhibited the development of monocrotaline-induced pulmonary hypertension, and this protective effect was blocked with the addition of tin protoporphyrin. In addition, treatment with cobalt protoporphyrin resulted in a substantial protection in this model of pulmonary hypertension. Rapamycin induction of HO-1 was dependent upon a transcriptional event; however, it was not mediated through an altered redox state or mammalian targets of rapamycin inhibition. Unlike wild-type cells, the growth of HO-1-/- mouse aortic smooth muscle cells was not inhibited or cell cycle arrested in G1 in response to rapamycin. This study demonstrates that HO-1 is critical for the antiproliferative and vascular protective effects of rapamycin in vitro and in vivo in monocrotaline-induced pulmonary hypertension. PMID- 16357869 TI - Positive and negative effects of widespread badger culling on tuberculosis in cattle. AB - Human and livestock diseases can be difficult to control where infection persists in wildlife populations. For three decades, European badgers (Meles meles) have been culled by the British government in a series of attempts to limit the spread of Mycobacterium bovis, the causative agent of bovine tuberculosis (TB), to cattle. Despite these efforts, the incidence of TB in cattle has risen consistently, re-emerging as a primary concern for Britain's cattle industry. Recently, badger culling has attracted controversy because experimental studies have reached contrasting conclusions (albeit using different protocols), with culled areas showing either markedly reduced or increased incidence of TB in cattle. This has confused attempts to develop a science-based management policy. Here we use data from a large-scale, randomized field experiment to help resolve these apparent differences. We show that, as carried out in this experiment, culling reduces cattle TB incidence in the areas that are culled, but increases incidence in adjoining areas. These findings are biologically consistent with previous studies but will present challenges for policy development. PMID- 16357870 TI - The Polycomb group protein EZH2 directly controls DNA methylation. AB - The establishment and maintenance of epigenetic gene silencing is fundamental to cell determination and function. The essential epigenetic systems involved in heritable repression of gene activity are the Polycomb group (PcG) proteins and the DNA methylation systems. Here we show that the corresponding silencing pathways are mechanistically linked. We find that the PcG protein EZH2 (Enhancer of Zeste homolog 2) interacts-within the context of the Polycomb repressive complexes 2 and 3 (PRC2/3)-with DNA methyltransferases (DNMTs) and associates with DNMT activity in vivo. Chromatin immunoprecipitations indicate that binding of DNMTs to several EZH2-repressed genes depends on the presence of EZH2. Furthermore, we show by bisulphite genomic sequencing that EZH2 is required for DNA methylation of EZH2-target promoters. Our results suggest that EZH2 serves as a recruitment platform for DNA methyltransferases, thus highlighting a previously unrecognized direct connection between two key epigenetic repression systems. PMID- 16357871 TI - Lgl, Pins and aPKC regulate neuroblast self-renewal versus differentiation. AB - How a cell chooses to proliferate or to differentiate is an important issue in stem cell and cancer biology. Drosophila neuroblasts undergo self-renewal with every cell division, producing another neuroblast and a differentiating daughter cell, but the mechanisms controlling the self-renewal/differentiation decision are poorly understood. Here we tested whether cell polarity genes, known to regulate embryonic neuroblast asymmetric cell division, also regulate neuroblast self-renewal. Clonal analysis in larval brains showed that pins mutant neuroblasts rapidly fail to self-renew, whereas lethal giant larvae (lgl) mutant neuroblasts generate multiple neuroblasts. Notably, lgl pins double mutant neuroblasts all divide symmetrically to self-renew, filling the brain with neuroblasts at the expense of neurons. The lgl pins neuroblasts show ectopic cortical localization of atypical protein kinase C (aPKC), and a decrease in aPKC expression reduces neuroblast numbers, suggesting that aPKC promotes neuroblast self-renewal. In support of this hypothesis, neuroblast-specific overexpression of membrane-targeted aPKC, but not a kinase-dead version, induces ectopic neuroblast self-renewal. We conclude that cortical aPKC kinase activity is a potent inducer of neuroblast self-renewal. PMID- 16357872 TI - Increased pituitary volume in ...psychosis study. PMID- 16357874 TI - Cost-effectiveness of losartan-based therapy in patients with hypertension and left ventricular hypertrophy: a UK-based economic evaluation of the Losartan Intervention for Endpoint reduction in hypertension (LIFE) study. AB - The Losartan Intervention for Endpoint reduction in hypertension (LIFE) study demonstrated the clinical benefit of losartan-based therapy in hypertensive patients with left ventricular hypertrophy (LVH), mainly due to a highly significant 25% reduction in the relative risk of stroke compared with an atenolol-based regimen, for a similar reduction in blood pressure. The aim of this economic evaluation was to estimate the cost-effectiveness of losartan compared with atenolol from a UK national health system perspective. Quality adjusted survival and direct medical costs were modelled beyond the trial using the within-trial incidence of stroke. Survival with stroke, study medication use and quality of life by stroke status were taken directly from the LIFE trial. The LIFE data were supplemented with UK data on lifetime direct medical costs of stroke and life expectancy in individuals without stroke. No additional stroke events or use of study treatment were assumed beyond the trial. Costs and benefits were discounted using current UK Treasury rates. In the base-case analysis, the reduction in stroke-related costs (by 968 sterling pound) offset 86% of the increase in study medication costs (1128 sterling pound) among losartan-treated patients. The incremental cost-effectiveness ratio (ICER) for losartan versus atenolol in hypertensive patients with LVH was 2130 sterling pound per quality-adjusted life year (QALY) gained (3195 Euro/QALY), and this increased to 11,352 sterling pound per QALY gained (16,450 Euro/QALY) when the costs of stroke beyond the first 5 years were excluded. Thus, the clinical benefit of losartan was achieved at a cost well within reported thresholds for cost-effectiveness. PMID- 16357876 TI - [Literature and medicine]. PMID- 16357875 TI - [More than medicine]. PMID- 16357877 TI - [Lead poisoning]. PMID- 16357878 TI - [Boy or girl?]. PMID- 16357881 TI - [What do we know about participation in cultural activities and health?]. AB - BACKGROUND: Knowledge of the association between health status and lifestyle factors, such as food habits, smoking and physical activity, is abundant. Other lifestyle factors, such as participation in cultural activities, have attained less attention. MATERIAL AND METHODS: The article is based on studies of the literature. Reference lists in key articles have been used as well as references given by research colleagues. RESULTS AND INTERPRETATION: The survey shows an association between participation in cultural activities, cultural experiences and health status, also when measured by differing methods. Further population studies, longitudinal studies and controlled studies are needed in order to expand our knowledge of the relationship between participation in cultural activities and health status. There is a need for multidisciplinary cooperation and increased use of combined quantitative and qualitative methods. PMID- 16357880 TI - [Why do some women only give birth to boys or to girls?]. AB - BACKGROUND: There are more families with many children of only one sex than can be explained by unconditional probability. MATERIAL AND METHODS: Data from the Medical Birth Registry of Norway, covering more than 540,000 women with two, three and four births for the period 1967-2003, were used to study how the sex distributions of children already born affected the probability of a new birth and the sex of the next sibling. RESULTS: Women with two children of the same sex had a higher probability of having more children compared to women with two children of both sexes (RR=1.14, (1.14-1.15)). This also applied to mothers with three children (RR=1.15 (1.13-1.17)). The probability was highest for mothers with boys only. Multiple births and parity affected the probability of giving birth to a boy versus a girl, but the sex composition of already born siblings had no influence. INTERPRETATION: We have no evidence that some parents may have a probability of having a boy versus a girl that differs from the rest of the population. The fact that there are more siblings of only one sex is mainly behavioural; some mothers with only girls or only boys keep having more children in an attempt to receive a child of the opposite sex. PMID- 16357882 TI - [Ingested lead cartridges cause trouble]. AB - BACKGROUND: Lead poisoning is rare. Intake of lead shots may cause damage of the bone marrow, the nervous system, the liver, the kidneys and the endocrine organs. MATERIALS AND METHODS: We present two patients who had taken approximately 120 grams of lead shots. They developed asthenia, nausea and abdominal pain. We tried to remove the lead shots with fluids, laxatives and colonoscopy, but ultimately surgical intervention had to be performed. The patients were also treated with dimercaptosuccinic acid (DMSA). RESULTS AND INTERPRETATION: Although DMSA treatment is associated with a significant decrease in blood lead concentration and an increase of lead urinary excretion, surgical intervention is the most efficient way of treating oral intake of lead. Treatment with DMSA will probably have to go on for months in order to give an additional effect. PMID- 16357883 TI - [Sexual assault against women-- injury pattern and victim-perpetrator relationship]. AB - BACKGROUND: Women who have been exposed to sexual assault have the opportunity to consult specialised health service. The Centre for victims of sexual assault at St. Olav University Hospital, Trondheim, Norway, offers emergency medical treatment, psychosocial follow-up and collects evidence in case the victim wants to file a complaint. The aim of this study was to examine whether injury pattern and assault characteristics differed according to the victim's relationship to the perpetrator. MATERIAL AND METHODS: Information about the assaults was collected retrospectively from medical journals of 162 female clients who presented at the centre over the period 1 June 2000 through 31 May 2003. Depending on their relationship with the perpetrator, the victims were categorised into four groups: stranger, accidental acquaintance, acquaintance, or partner. Assault characteristics and injury patterns were compared among these groups. RESULTS: 69 % (111/162) of the victims knew their offender. Accidental acquaintances were reported as perpetrator in 32 (20 %) of the assaults, acquaintances in 68 (42 %), partners in 11 (7 %) and unknown offenders in 29 (18 %). Type and severity of the sexual assaults did not differ significantly according to victim-perpetrator relationship. The most life-threatening violence and use of a weapon were reported used by known offenders only. INTERPRETATION: In emergency medical response to victims of sexual assaults, it should be made sure that victims, independently of their relationship with perpetrator, are met and treated equally. PMID- 16357884 TI - [Use and misuse of albumin]. PMID- 16357885 TI - [Cultural approaches to treatment and public health work]. AB - BACKGROUND: Improvement of health status due to physical activity is well documented. Research on health effects of other cultural measures is more scarce. During recent years, considerable work in these fields has been presented in the Nordic countries and there is a good basis for further research. MATERIAL: The article is a literature study based on reports and literature from projects in which cultural measures have been used as elements in treatment, rehabilitation, preventive and health promotion in the Nordic countries, primarily in Norway. Prominent workers within cultural and health work in the Nordic countries have contributed with information on relevant literature and reports. RESULTS AND INTERPRETATION: Reports from projects in Norway on culture and health indicate an improvement on individual life quality and experienced health status. Lack of research leaves an uncertainty as to whether the effects are lasting and whether other measures would have had similar effects. International research literature indicates that cultural activities may have health effects. There is a need for further research based on multidisciplinary approaches. PMID- 16357886 TI - [Norwegian physicians' knowledge of Latin medical terminology]. AB - BACKGROUND: It is well established that medical terminology is firmly based in Latin and Greek. However, teaching in Latin and medical terminology is not adequate in the medical education in Norway. MATERIAL AND METHODS: A questionnaire on medical practitioners' knowledge of medical Latin was handed out among doctors participating in further education at the University of Tromso, spring 2005. The questionnaire was divided into three parts: knowledge of Latin medical words, Latin names on diseases, and grammar (Latin word endings). The response rate was 102/121 (84 %).; 102 doctors participated, including 51 general practitioners. RESULTS: The understanding of Latin medical words was generally good. 70 % (71/102) of the doctors had less than three errors. The knowledge of Latin names for diseases was also quite good, with 73 % (74/102) having less than three errors. However, there were many grave mistakes. The ability to give correct word endings in plural nominative and singular genitive was almost absent; 85 % (87/102) made four or more errors. INTERPRETATION: Although Norwegian medical practitioners have a certain degree of skills in medical Latin, their knowledge is not sufficient. Marginal knowledge of important medical terms may complicate communication between doctors. A compulsory minimum of training in medical nomenclature should be a part of the curriculum in medical schools. PMID- 16357887 TI - [The individual stories and the professional conversations]. PMID- 16357888 TI - [Mutual respect better than careful goodness]. PMID- 16357890 TI - [Health for all possible without democracy?]. PMID- 16357891 TI - [Lars Hertervig and Carl Fredrik Hill--creativeness and mental illness]. AB - It seems to be a correlation between creativity and mental illnesses like manic depressive illness and schizophrenia. The reason for this is partly unknown, but neuropsychological investigations have proposed mechanisms which could help to explain this association. Lars Hertervig (1830-1902) and Carl Fredrik Hill (1849 1911) are two famous Scandinavian landscape painters from the end of the 19th century, both of whom suffered from mental illness. This paper describes the artistic development of the two painters and discusses this in relation to how brain diseases may influence creativity and artistic expression. PMID- 16357892 TI - [Descriptions of mental illness in Alf Proysen's first book]. AB - One of Norway's most famous artists after the Second World War was Alf Proysen (1914-70). He made his debut with a collection of short stories in 1945 entitled "Dorstokken heme" (The doorsill at home). Three of the short stories deal with women who develop mental illness. This article is about how Proysen describes the mental illnesses and what might have provoked them. PMID- 16357893 TI - [Louis-Ferdinand Celine--physician, writer, anti-semite]. PMID- 16357894 TI - [The physician, classical literature and the psychiatric patient]. AB - The physician and in particular the psychiatric resident is regularly confronted by the humanistic dimension of psychiatry. Impressions from classical literature can be seen as a special inspiration and an enriching factor for acquiring understanding. The literary classics represent the earliest "documentation" of the psychology and psychopathology of humankind. The unconscious, irrational and enigmatical aspect of human existence and its aberrations is a shared interest in literature, psychology and medicine. Fictional literature conveys contributions of relevance to the history of psychiatry, on mental institutions, doctors' attitudes, ethical dilemmas as well as descriptions and thoughts about mental states. The borderland between normality and insanity is often a subject of particular interest. Literary classics represent a "transcending psychology" in contrast to other prevailing scientific categories and delineations. The language of the author generally implies a challenging, non-theoretical alternative, springing from the individual and his or her unique destiny. The language is usually non-moralising, frequently with a surprising and wondering use of lucid images and allegories. Classical world literature is predominantly referred to: Goethe, Kleist and Turgenev as portrayers of personality disorders; Dostoyevsky, Shakespeare and Nerval on psychoses, and the Old Testament and Snorri Sturlasson on affective disorders. PMID- 16357895 TI - [Physicians in the TV soap--a study of ER]. AB - Health providers are constantly exposed to patients' cultural expressions. Television is a central agent forming our cultural surroundings. The aim of this study has been to elucidate how a specific genre - the medical soap opera - represents the health system, and shape our expectations of it. The first part of the study is a genre analysis based on media science, literary theory and narrative-based medicine. The second part of the study applies these theoretical elements on the series Emergency Room (ER). Using text analysis, one episode was reviewed in detail. The medical soap opera is a genre characterised by qualities such as a narration, its focus on the physician in her professional role and as a private individual, and by a glamour medical reality. These theoretical considerations support how soap operas focus on the physician as a type and a causally stringent construction of the narrative. The narrative qualities of the soap opera are found to satisfy people's desire for more causality in daily life. Wish fulfillment is a central function of the soap opera. In sum, medical soap operas fulfill a desire for a more comprehensive health system, showing health actors as human beings rather than just white coats. PMID- 16357896 TI - [Infant mortality in Asker and Baerum in the 18th and 19th century]. AB - BACKGROUND: The decline in infant mortality is an important part of the secular decline in mortality in the western world. The major causes of the decline are subject to controversy. MATERIAL AND METHODS: Individual event records from censuses, church records and land registers from two Norwegian parishes during the years 1814-1878 were registered and linked into individual life course records. Around 15,000 infants, of whom 1500 died, were analysed in depth with Cox regression analysis. The total yearly counts of births and infant deaths from 1735 were analysed using ordinary linear regression. RESULTS: Infant mortality hovered around 23 per cent during the middle of the 18th century and fell to a level around 10 per cent by the end of the 19th century. The decline was strongest during the neonatal period. Women born during the first decade of the 19th century, a decade known for a succession of years with bad harvests, war and high infant mortality, gave birth to infants with increased neonatal mortality. INTERPRETATION: The decline in infant mortality during the first part of the 19th century can thus be attributed to an improvement in the health of the mothers dating back to their own fetal or infant stage. The decline took place in the absence of trained medical personnel. PMID- 16357897 TI - [Eugenic indication for abortion--a historical background]. AB - BACKGROUND: Modern discussions on fetal diagnostics and abortion frequently leave the impression that the eugenic clause in the Norwegian abortion act is a heritage from the eugenics movement in the 1930s. However, although much has been written on this, few have systematically pursued the historical development of the eugenic clause. MATERIAL AND METHODS: The article describes the development of Norwegian abortion legislation in general, and the eugenic clause in particular, from the discussions in the 1930s, through the Nazi abortion law during the German occupation (which is relatively unknown in the historiographic literature), and to the legislation that was finally passed after the war. The sources are primarily official documents (committee reports, drafts for legislation, debates in parliament) as well as discussions in the Journal of the Norwegian Medical Association. Various proposals are discussed according to two criteria: a) the role hereditary arguments played in the formulation of a eugenic clause; and b) the extent to which such a clause is justified by social, as opposed to individual, concerns. RESULTS AND INTERPRETATION: Although social concerns every now and then figure in justifications for the eugenic clause, the lack of eugenic argumentation in Norwegian abortion debates until 1960 is striking. There are traces of a clear eugenic or racial hygienic approach in the 1930s, but this thinking was left dead by the abortion act instituted by the Nazi regime. PMID- 16357898 TI - [Was Spanish influenza in Kristiania a socially neutral disease?]. AB - BACKGROUND: A debate has been going on since 1918 over whether there were social differences in mortality from Spanish influenza. Mot commentators have concluded that mortality was socially neutral. In this paper it is discussed why the influenza pandemic of 1918-19 was different from recent influenza epidemics, which have displayed greater social differences in mortality than most other causes of death. MATERIALS AND METHODS: Multivariate analysis and event history data are used for the first time to analyse the variation in mortality from Spanish influenza. The study is confined to a west end and an east end parish in Norway's capital city. Historical demographic data for Norway has a good reputation internationally. The data are also unique because--Norway being a neutral country--they can be used to estimate the effect of the Spanish influenza on mortality net of the effect of the First World War. RESULTS: Middle class and bourgeois mortality was 19-25% lower than working class mortality (not significant). Those living in apartments with four to six rooms had on average 50 % lower mortality than those living in one-room apartments (significant). Moreover, the east end parish had 50 % higher mortality than the west end parish (significant). INTERPRETATION: These findings challenge the myth that Spanish influenza picked its victims at random. The study may have international relevance because data difficult to obtain for other countries have been used. PMID- 16357899 TI - [The drinking problem--temperance after prohibition]. AB - The alcohol issue had a prominent place in Norwegian politics and society in the first decades of the 20th century; teetotalers' organisations had a major role. Prohibition was the ultimate way of solving the alcohol problem. After the prohibition (1916-27), the teetotalers lost their hegemony. Anti-prohibition views became influential in the making of Norwegian alcohol policy. The post prohibition years have been seen as at period of change in which a moralist and condemning attitude to alcohol and alcoholics were replaced by a more humane disease concept of alcoholism. This article challenges this view. After prohibition, to separate between use and abuse of alcohol was the main focus of Norwegian alcohol policy. Alcohol abuse and alcohol abusers became the main targets. Abuse of alcohol was first and foremost seen as a social category, as immoral behaviour. In the 1930s, a moralist and condemning attitude toward excessive drinking and drinkers was put forward, not replaced, in Norwegian alcohol policy. A concept of alcoholism as a disease had little or no impact in these years. PMID- 16357900 TI - [Occupational lead poisoning 1892-1936]. AB - The first Norwegian factory inspection act was passed in 1892. Until 1931, little attention was paid to occupational lead poisoning. In 1915 the physician Olai Lorange (1876-1965) was appointed chief of the factory inspectorate. In 1916 he initiated a registration of lead poisoning in Norway. In 1920 he also called on Norwegian doctors to report diseases considered to be of occupational origin. The results were scanty. Attention was paid to occupational lead poisoning in a draft legislation put forward in 1923. However, the motion was not put to vote during the depression after the First World War and was not passed until 1936. Historically, lead exposure has in periods been forgotten as a cause of disease, only to reappear. In Norway, occupational lead poisoning attracted attention in 1931-33, when 46 cases were reported from a shipyard. The inspectorate laid down regulations and the government granted compensations and pensions to 40 of the workers. Based on worker protection acts and regulations, yearbooks from the factory inspectorate, and medical literature, the article describes attitudes towards occupational lead poisoning in the early years after the first Norwegian factory inspection act was implemented. PMID- 16357901 TI - [Cholera and quarantine in Kristiansand]. AB - BACKGROUND AND MATERIAL: Cholera was the horror disease in Norwegian cities in the 18th century. The City of Kristiansand had a quarantine station and hospital at Odderoy island from 1804. We have studied the quarantine organisation and compared the number of cholera deaths in different cities along the coast. RESULTS: Drammen is comparable to Kristiansand and had many visiting trading ships. During 1832-33, 1833 and 1857 the Drammen community had registered 544 cholera patients; 336 died. Kristiansand had a total of only 15 deaths (1833, 1853 and 1866), the quarantine hospital included. Also compared to other cities, Kristiansand had few deaths. INTERPRETATION: The reason for the better cholera statistics in Kristiansand is probably the quarantine station and hospital at Odderoy island (1804). The city's health authorities had efficient routines and knowledge, probably adopted from the quarantine station and the hospital. PMID- 16357902 TI - [Neurosurgery in antique medicine]. AB - Trepanation and craniotomy are two of the oldest surgical procedures known, and extensive archaeological evidence of trepanation exists in ancient cultures. However, the first descriptions of the surgical techniques are from Greek and Roman medicine, where cranial surgery was used to treat head trauma. This article concerns neurosurgery in ancient medicine, with an emphasis on "De Medicina" by Aulus Cornelius Celsus (25 BC-50 AD) and the Corpus Hippocraticum (about 400 BC). These texts are further considered in the light of excavated surgical instruments from Pompeii. PMID- 16357903 TI - [Lorentz Eldjarn--a catalyst for chemistry in Norwegian medicine]. AB - Lorentz Eldjarn, MD (born 1920) was professor of clinical biochemistry at the University of Oslo. He modernised clinical chemistry both nationally and internationally. He introduced both quality control and mass spectrometry in clinical chemistry. In the 1950s he studied the radioprotective compound cystamine and showed that it forms mixed disulfides with protein SH groups. In part, this explains its radioprotective effect. In 1961, he founded the Department of Clinical Biochemistry at the University of Oslo, where most of the Norwegian clinical chemists were trained in the following years. His introduction of mass spectrometry as an analytical tool in clinical chemistry led to the detection of three new errors of metabolism in his institute: methylmalonic aciduria, beta-hydroxyisovaleric aciduria, and pyroglutamic aciduria. In 1975, he was awarded the "Distinguished Clinical Chemist Award" of the International Association of Clinical Chemists, and in 1976 the Scandinavian Jahre research prize. After he resigned from his university position in 1978, he has continued his work to develop stable standard sera for clinical chemistry. PMID- 16357904 TI - [Paul A. Owren--a 100 years' anniversary]. PMID- 16357905 TI - [Norwegian medical students in German prisons]. PMID- 16357906 TI - [Norwegian medical students in Denmark 1945-55]. PMID- 16357907 TI - [Reproduction, rights and ethics]. PMID- 16357908 TI - [District medicine]. PMID- 16357909 TI - [I wish 1 000 new general practitioners for Christmas]. PMID- 16357910 TI - [Misleading about type 2 diabetes and insulin]. PMID- 16357912 TI - [Lysenkoism in Norwegian nutrition science?]. PMID- 16357914 TI - [Monitoring of fetal movements--documentation is insufficient]. PMID- 16357917 TI - [Stroke patients and vision]. PMID- 16357918 TI - [The seroxat issue]. PMID- 16357920 TI - [Treatment of patients with acute coronary syndrome in Norwegian hospitals]. PMID- 16357922 TI - [The Medical Society and medicalization of forensic medicine]. PMID- 16357926 TI - An introduction to health impact assessment. PMID- 16357927 TI - Contemporary debates in health impact assessment: What? Why? When? PMID- 16357928 TI - Response to 'Contemporary debates in health impact assessment'. PMID- 16357929 TI - Health impact assessment: an international perspective. PMID- 16357930 TI - Health impact assessment in Australia. PMID- 16357931 TI - Health impact assessment in New Zealand. PMID- 16357932 TI - The art of the possible: experience and practice in health impact assessment in New South Wales. PMID- 16357933 TI - Building an equity focus in health impact assessment. PMID- 16357934 TI - The NSW Health impact assessment project. PMID- 16357935 TI - Constructing a database of development applications considered by public health units in NSW. PMID- 16357936 TI - Health impact assessment case study: working with local government fo obtain health benefits. PMID- 16357937 TI - Health impact assessment on an integrated chronic disease prevention campaign. PMID- 16357938 TI - NSW health Aboriginal health impact statement. PMID- 16357939 TI - Communicable Diseases Report, May and June 2005. PMID- 16357940 TI - SOCS1 restricts dendritic cells' ability to break self tolerance and induce antitumor immunity by regulating IL-12 production and signaling. AB - DC-based tumor vaccine research has largely focused on enhancing DC maturation/costimulation and antigen presentation in order to break tolerance against self tumor-associated antigens. DC immunization can activate autoreactive T cells but rarely causes autoimmune pathologies, indicating that self tolerance at the host level is still maintained in the vaccinated hosts. This study in mice reveals a novel regulatory mechanism for the control of self tolerance at the host level by DCs through the restriction of positive cytokine feedback loops by cytokine signaling inhibitor SOCS1. The study further finds the requirement of persistent antigen presentation by DCs for inducing pathological autoimmune responses against normal tissues and tumor, which can be achieved by silencing SOCS1 to unleash the unbridled signaling of IL-12 and the downstream cytokine cascade. However, the use of higher-affinity self peptides, enhancement of DC maturation, and persistent stimulation with cytokines or TLR agonists fail to break tolerance and induce pathological antitumor immunity. Thus, this study indicates the necessity of inhibiting SOCS1, an antigen presentation attenuator, to break self tolerance and induce effective antitumor responses. PMID- 16357941 TI - Intramuscular viral delivery of paraplegin rescues peripheral axonopathy in a model of hereditary spastic paraplegia. AB - Degeneration of peripheral motor axons is a common feature of several debilitating diseases including complicated forms of hereditary spastic paraplegia. One such form is caused by loss of the mitochondrial energy-dependent protease paraplegin. Paraplegin-deficient mice display a progressive degeneration in several axonal tracts, characterized by the accumulation of morphological abnormal mitochondria. We show that adenoassociated virus-mediated (AAV-mediated) intramuscular delivery of paraplegin halted the progression of neuropathological changes and rescued mitochondrial morphology in the peripheral nerves of paraplegin-deficient mice. One single injection before onset of symptoms improved the motor performance of paraplegin-deficient mice for up to 10 months, indicating that the peripheral neuropathy contributes to the clinical phenotype. This study provides a proof of principle that gene transfer may be an effective therapeutic option for patients with paraplegin deficiency and demonstrates that AAV vectors can be successfully employed for retrograde delivery of an intracellular protein to spinal motor neurons, opening new perspectives for several hereditary axonal neuropathies of the peripheral nerves. PMID- 16357942 TI - A new type of radiosensitive T-B-NK+ severe combined immunodeficiency caused by a LIG4 mutation. AB - V(D)J recombination of Ig and TCR loci is a stepwise process during which site specific DNA double-strand breaks (DSBs) are made by RAG1/RAG2, followed by DSB repair by nonhomologous end joining. Defects in V(D)J recombination result in SCID characterized by absence of mature B and T cells. A subset of T-B-NK+ SCID patients is sensitive to ionizing radiation, and the majority of these patients have mutations in Artemis. We present a patient with a new type of radiosensitive T-B-NK+ SCID with a defect in DNA ligase IV (LIG4). To date, LIG4 mutations have only been described in a radiosensitive leukemia patient and in 4 patients with a designated LIG4 syndrome, which is associated with chromosomal instability, pancytopenia, and developmental and growth delay. The patient described here shows that a LIG4 mutation can also cause T-B-NK+ SCID without developmental defects. The LIG4-deficient SCID patient had an incomplete but severe block in precursor B cell differentiation, resulting in extremely low levels of blood B cells. The residual D(H)-J(H) junctions showed extensive nucleotide deletions, apparently caused by prolonged exonuclease activity during the delayed D(H)-J(H) ligation process. In conclusion, different LIG4 mutations can result in either a developmental defect with minor immunological abnormalities or a SCID picture with normal development. PMID- 16357943 TI - Controlled delivery of PDGF-BB for myocardial protection using injectable self assembling peptide nanofibers. AB - Endothelial cells can protect cardiomyocytes from injury, but the mechanism of this protection is incompletely described. Here we demonstrate that protection of cardiomyocytes by endothelial cells occurs through PDGF-BB signaling. PDGF-BB induced cardiomyocyte Akt phosphorylation in a time- and dose-dependent manner and prevented apoptosis via PI3K/Akt signaling. Using injectable self-assembling peptide nanofibers, which bound PDGF-BB in vitro, sustained delivery of PDGF-BB to the myocardium at the injected sites for 14 days was achieved. A blinded and randomized study in 96 rats showed that injecting nanofibers with PDGF-BB, but not nanofibers or PDGF-BB alone, decreased cardiomyocyte death and preserved systolic function after myocardial infarction. A separate blinded and randomized study in 52 rats showed that PDGF-BB delivered with nanofibers decreased infarct size after ischemia/reperfusion. PDGF-BB with nanofibers induced PDGFR-beta and Akt phosphorylation in cardiomyocytes in vivo. These data demonstrate that endothelial cells protect cardiomyocytes via PDGF-BB signaling and that this in vitro finding can be translated into an effective in vivo method of protecting myocardium after infarction. Furthermore, this study shows that injectable nanofibers allow precise and sustained delivery of proteins to the myocardium with potential therapeutic benefits. PMID- 16357944 TI - Early G2/M checkpoint failure as a molecular mechanism underlying etoposide induced chromosomal aberrations. AB - Topoisomerase II (Topo II) inhibitors are cell cycle-specific DNA-damaging agents and often correlate with secondary leukemia with chromosomal translocations involving the mixed-lineage leukemia/myeloid lymphoid leukemia (MLL) gene on chromosome 11 band q23 (11q23). In spite of the clinical importance, the molecular mechanism for this chromosomal translocation has yet to be elucidated. In this study, we employed 2-color FISH and detected intracellular chromosomal translocations induced by etoposide treatment. Cells such as ataxia telangiectasia mutated-deficient fibroblasts and U2OS cells, in which the early G2/M checkpoint after treatment with low concentrations of etoposide has been lost, executed mitosis with etoposide-induced DNA double-strand breaks, and 2 color FISH signals located on either side of the MLL gene were segregated in the postmitotic G1 phase. Long-term culture of cells that had executed mitosis under etoposide treatment showed frequent structural abnormalities of chromosome 11. These findings provide convincing evidence for Topo II inhibitor-induced 11q23 translocation. Our study also suggests an important role of the early G2/M checkpoint in preventing fixation of chromosomal abnormalities and reveals environmental and genetic risk factors for the development of chromosome 11 translocations, namely, low concentrations of Topo II inhibitors and dysfunctional early G2/M checkpoint control. PMID- 16357945 TI - Development of a miniaturized assay for the high-throughput screening program for poly(ADP-ribose) polymerase-1. AB - Poly(ADP-ribose) polymerase (PARP) plays a pivotal role in the repair of DNA strand breaks. However, excessive activation of PARP causes a rapid depletion of intracellular energy, leading to cell death. PARP inhibitors may have potential therapeutic benefit in the treatment of myocardial ischemia, stroke, and neurodegenerative disease. With these emerging medicinal interests, various screening programs have identified small molecules that inhibit PARP with reasonable potencies. However, the increasing numbers of diverse small molecules generated through combinatorial chemistry necessitate the use of robust assays with good sensitivity and specificity for use as a high-throughput screening (HTS) program. Here, we report the development and the validation of a nonisotopic PARP-1 assay suitable for HTS by converting a biotinylated NAD-based colorimetric assay to a miniaturized 384-well plate format. Comparing with the conventional methods, this miniaturized PARP-1 inhibition assay was equally sensitive with excellent reproducibility and cost-effectiveness. Because nonisotopic PARP-1 inhibition assays are widely used, the methodology described in this article can expand the feasibility of this assay as a high-throughput assay for screening of PARP-1 inhibitors from a random chemical library. PMID- 16357946 TI - Testosterone induced Ca2+ influx in bone marrow-derived macrophages via surface binding sites. AB - The biological activity of testosterone is thought to occur predominantly through binding to the androgen receptor (AR), a member of the nuclear receptor superfamily that functions as a ligand-activated transcription factor. Here, we found that testosterone could induce a rapid rise in the intracellular free Ca2+ concentration ([Ca2+]i) of Fura-2 loaded bone marrow-derived macrophages (BMMs), which was found to be predominantly due to the influx of extracellular Ca2+ through Ni2+-blockable Ca2+ channels in the plasma membrane. However, these effects of testosterone could not be associated with the classical intracellular AR in BMMs, since AR was not detectable using different experimental techniques. Instead, it was found that testosterone could bind to the surface of BMMs by the use of an impermeable testosterone-BSA-FITC, and Ca2+ influx could also be induced by testosterone conjugated to BSA. Our data indicated a novel mode of direct action of testosterone on BMMs, which was not mediated through the classical AR response, but through the binding sites of testosterone on cell surfaces. PMID- 16357947 TI - Ondansetron does not block tramadol-induced analgesia in mice. AB - Tramadol is a weak opioid agonist and an inhibitor of the reuptake of noradrenaline and serotonin. This study was undertaken to assess a possible pharmacological interaction of ondansetron, a serotonin-3 (5-hydroxytryptamine-3, 5-HT3) antagonist, and tramadol in an animal model for acute pain. Sixty-three male albino mice were randomly given saline, tramadol (10, 20, and 40 mg kg(-1)), ondansetron (1, 2, and 4 mg kg(-1)), or ondansetron (1, 2, and 4 mg kg(-1)) and tramadol (20 mg kg(-1), given 10 min after ondansetron injection) intraperitoneally. Each mouse was assessed twice for tail-flick latency before saline or drug administration and 15, 30, 60, 90, and 120 min thereafter. Tramadol (10 mg kg(-1)) had no effect on pain threshold levels of mice, but tramadol doses of 20 or 40 mg kg(-1) increased pain threshold levels in a dose dependent manner (p < 0.01 for 20 mg kg(-1) and p < 0.001 for 40 mg kg(-1)). Ondansetron doses of 1, 2, or 4 mg kg(-1) alone had no effect on pain threshold levels of mice. Tramadol (20 mg kg(-1)) and ondansetron (1, 2, and 4 mg kg(-1)) increased pain threshold levels at all doses (p < 0.001 for 1 and 2 mg kg(-1) ondansetron and p < 0.01 for 4 mg kg(-1) ondansetron). The pain threshold levels of mice given tramadol (20 mg kg(-1)) alone or tramadol and ondansetron (p > 0.05 for 1, 2, and 4 mg kg(-1)) were similar. Our results indicate that ondansetron-a 5-HT3 selective antagonist-does not decrease the analgesic effectiveness of tramadol in mice, which may be the result of different mechanisms involving 5-HT3 receptors. PMID- 16357948 TI - Antilithiatic effect of Asparagus racemosus Willd on ethylene glycol-induced lithiasis in male albino Wistar rats. AB - The ethanolic extract of Asparagus racemosus Willd. was evaluated for its inhibitory potential on lithiasis (stone formation), induced by oral administration of 0.75% ethylene glycolated water to adult male albino Wistar rats for 28 days. The ionic chemistry of urine was altered by ethylene glycol, which elevated the urinary concentration of crucial ions viz. calcium, oxalate, and phosphate, thereby contributing to renal stone formation. The ethanolic extract, however, significantly (p < 0.05) reduced the elevated level of these ions in urine. Also, it elevated the urinary concentration of magnesium, which is considered as one of the inhibitors of crystallization. The high serum creatinine level observed in ethylene glycol-treated rats was also reduced, following treatment with the extract. The histopathological findings also showed signs of improvement after treatment with the extract. All these observations provided the basis for the conclusion that this plant extract inhibits stone formation induced by ethylene glycol treatment. PMID- 16357949 TI - Effect of cefepime on startle amplitude and prepulse inhibition of startle reflex in rats: manipulation of GABAA receptor function with midazolam. AB - Disruption of prepulse inhibition (PPI) of startle response is manifested in schizophrenia and Huntington's disease among other neuropsychiatric disorders characterized by sensorimotor gating deficit. Antagonism of GABAA receptor function was documented to reduce PPI response. However, whether GABAA antagonism reduces PPI response is yet to be established because of contradicting reports. Cefepime is one of the fourth-generation cephalosporins documented to antagonize GABAA receptor function. This study investigated the effect of intramuscular injection of cefepime (45 and 90 mg/kg) twice daily for three consecutive days on both PPI of acoustic startle response and acoustic startle amplitude in rats. The effect of administration of the GABAA receptor agonist midazolam (1 mg/kg, i.p.) in conjunction with cefepime on PPI response and startle amplitude was also investigated. Results showed that administration of both dose levels of cefepime caused PPI deficit. Treatment of animals with midazolam in conjunction with cefepime reversed the effect of the lower dose, but not the higher one, on PPI of startle response without affecting startle amplitude in both dose levels. Results of this study, therefore, support the view that GABAA antagonism reduces PPI response. It is also concluded that antagonism of GABAergic transmission may be involved in the effect of the lower dose of cefepime on PPI response. Other mechanisms may mediate the effect of the higher dose of cefepime on PPI response. Clinical investigations are needed to determine the consequence of using cefepime in disorders of sensorimotor gating. PMID- 16357950 TI - Involvement of dopamine D3 and D4 receptors in the discriminative stimulus properties of cocaine in the rat. AB - The present study was carried out to determine the involvement of dopamine receptor subtypes D3 and D4, in the discriminative stimulus effects of cocaine in the rats trained to discriminate 10 mg/kg of cocaine from vehicle. The discriminative stimulus effects of cocaine (1-10 mg/kg) were dose-dependent. The dopamine D2 receptor agonist bromocriptine (1.25-20 mg/kg) and the dopamine D3 receptor agonist R(+)-7-OH-DPAT (0.0001-0.3 mg/kg) produced cocaine (10 mg/kg) like discriminative stimulus effects. Both the dopamine D3 receptor antagonist GR103691 (1 mg/kg) and the dopamine D4 receptor antagonist L745870 (1 mg/kg) partially antagonized the discriminative stimulus effects of cocaine (10 mg/kg) and the cocaine (10 mg/kg)-like discriminative stimulus effects of R(+)-7-OH-DPAT (0.3 mg/kg). L745870 (0.001 mg/kg) inhibited the antagonistic effects of GR103691 (1 mg/kg) on the discriminative stimulus effects of cocaine (10 mg/kg), whereas the drug (0.001 mg/kg) enhanced the antagonistic effects of GR103691 (1 mg/kg) on the cocaine (10 mg/kg)-like discriminative stimulus effects of R(+)-7-OH-DPAT (0.3 mg/kg). GR103691 (1 mg/kg) in combination with L745870 (0.001 mg/kg) did not markedly affect the cocaine (10 mg/kg)-like discriminative stimulus effects of bromocriptine (20 mg/kg). These results suggest that the discriminative stimulus effects of cocaine are different from the cocaine-like discriminative stimulus effects of bromocriptine or R(+)-7-OH-DPAT, in terms of dopamine D3 and D4 receptors. PMID- 16357951 TI - Warming and response to contractile agents in calf cardiac vein: role of the Ca2+, KCa2+, and Na+ ion channel blockers. AB - The effects of warming (to 41 degrees C) on the serotonin (5-HT, 10(-8)-3 x 10( 3) M)- and carbachol (10(-)9-3 x 10(-4) M)-induced contractions and the role of calcium (Ca2+), potassium (K+), and sodium (Na+) channel blockers, in the warming induced responses were investigated in the calf cardiac vein. Concentration response curves to 5-HT and carbachol were isometrically recorded at 37 and 41 degrees C (control). The same procedure was repeated at 41 degrees C in the presence of verapamil (10(-6) M), caffeine (3 x 10(-4) M), tetraethylammonium (TEA, 10(-3)M), flecainide (10(-6) M), and also in the Ca2+-free medium with ethylene glycol bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA). During warming, the sensitivity, but not the maximal response, was significantly higher. Warming to 41 degrees C after treatment with verapamil or flecainide decreased the sensitivity, whereas treatment with caffeine increased the sensitivity significantly. Treatment with TEA did not modify the effect of warming. Furthermore, warming to 41 degrees C after incubation in Ca2+-free solutions with EGTA decreased the sensitivity to 5-HT and carbachol. The results of this study suggest the role for Ca2+ and Na+ ions in the warming-induced changes of cardiac vein treated with 5-HT and carbachol. PMID- 16357952 TI - Effects of single and long-term theophylline treatment on the threshold of mechanical nociception: contribution of adenosine A1 and alpha2-adrenoceptors. AB - The effects of single and long-term treatment with theophylline as well as the influence of adenosine A1 receptor agonist cyclopentyladenosine (CPA) and a adrenergic receptor antagonists prazosin and yohimbine were assessed in the paw pressure test in rats. Both single (37.5 and 75 mg/kg) and long-term (75 mg/kg/day, 14 days i.p.) theophylline treatments exerted antinociceptive effect by increasing the mechanical pain threshold. Single treatment of theophylline (75 mg/kg) antagonized the antinociceptive effect of CPA (0.1 mg/kg); CPA (0.1 mg/kg) abolished the theophylline-induced antinociception. Chronic treatment with theophylline did not change the antinociceptive effect of CPA, while CPA decreased the theophylline antinociception. Yohimbine (0.5 mg/kg), an a 2 adrenoceptor antagonist, diminished the antinociception of a single dose (75 mg/kg) of theophylline, whereas prazocin, an a 1-adrenoceptor antagonist, did not affect it. These results suggest that adenosine A1 and a 2-adrenoceptors take part in the antinociception induced by a single dose of theophylline. The antinociception induced by chronic theophylline treatment probably has a more complex mechanism in which the involvement of adenosine A1. PMID- 16357953 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials are a guide to the most recent clinical trials in current literature and congresses. The data in the following tables have been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity(R), the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: (Z)-4-Hydroxytamoxifen, [18F]-FPS; Adalimumab, alefacept, alemtuzumab, alfimeprase, aprepitant, aripiprazole, atomoxetine hydrochloride; Belatacept, bortezomib; C340, caspofungin acetate, clazosentan sodium, Cypher; Darbepoetin alfa, DB-289, decitabine, dronedarone hydrochloride, duloxetine hydrochloride; Eletriptan, entecavir, ertapenem sodium, escitalopram oxalate, eszopiclone, etoricoxib; Gaboxadol, gadofosveset sodium, galiximab, gemifloxacin mesilate, glutamine; Human insulin; I-131 ch-TNT-1/B, indiplon, inhaled insulin, isatoribine; L-Arginine hydrochloride, liposomal doxorubicin, lopinavir/ritonavir, lumiracoxib; Magnesium sulfate; Natalizumab; Olmesartan medoxomil, omapatrilat, OncoVEX (GM-CSF); rDNA insulin, rupatadine fumarate; Sorafenib; Tadalafil, teduglutide, temsirolimus, tenofovir disoproxil fumarate, tiotropium bromide; Valdecoxib, vardenafil hydrochloride hydrate. PMID- 16357956 TI - Synthesis of anionic methylpalladium complexes with phosphine-sulfonate ligands and their activities for olefin polymerization. AB - Reaction of diarylphosphinobenzene-2-sulfonic acids with tertially amines, followed by addition of [PdMeCl(cod)], provided anionic methylpalladium(II) complexes with bidentate phosphine-sulfonate ligands, which show high activity for copolymerization of ethylene with methyl acrylate. PMID- 16357957 TI - Dihydrobis(methimazolyl)borate and methimazolyl complexes of titanium. AB - The first poly(methimazolyl)borato complex of group 4, [Ti([double bond, length as m-dash]NCMe(3)){H(2)B(mt)(2)}(2)](mt = methimazolyl), results from the reaction of Na[H(2)B(mt)(2)] with [Ti([double bond, length as m dash]NCMe(3))Cl(2)(py)(3)] and features both kappa(2)-S,S' and kappa(3)-H,S,S' coordination of H(2)B(mt)(2) ligands coincident within the same molecule. PMID- 16357958 TI - Lanthanide(III) and group 13 metal ion complexes of tripodal amino phosphinate ligands. AB - The tripodal amino-phosphinate ligands, tris(4-(phenylphosphinato)-3-benzyl-3 azabutyl)amine (H(3)ppba.2HCl.H(2)O) and tris(4-(phenylphosphinato)-3 azabutyl)amine (H(3)ppa.HCl.H(2)O) were synthesized and reacted with Al(3+), Ga(3+), In(3+) and the lanthanides (Ln(3+)). At 2 : 1 H(3)ppba to metal ratios, complexes of the type [M(H(3)ppba)(2)](3+)(M = Al(3+), Ga(3+), In(3+), Ho(3+) Lu(3+)) were isolated. The bicapped [Ga(H(3)ppba)(2)](NO(3))(2)Cl.3CH(3)OH was structurally characterized and was shown indirectly by various techniques to be isostructural with the other [M(H(3)ppba)(2)](3+) complexes. Also, at 2 : 1 H(3)ppba to metal ratios, complexes of the type [M(H(4)ppba)(2)](5+)(M = La(3+) Tb(3+)) were characterized, and the X-ray structure of [Gd(H(4)ppba)(2)](NO(3))(4)Cl.3CH(3)OH was determined. At 1 : 1 H(3)ppba to metal ratios, complexes of the type [M(H(4)ppba)](4+)(M = La(3+)-Er(3+)) were isolated and characterized. Elemental analysis and spectroscopic evidence supported the formation of a 1 : 1 monocapped complex. Reaction of 1 : 1 ratios of H(3)ppa with Ln(3+) and In(3+) yielded complexes of the type [M(H(3)ppa)](3+)(M = La(3+) Yb(3+)) but with Ga(3+), complex of the type [Ga(ppa)].3H(2)O was obtained. Reaction of 1 : 1 ratios of H(3)ppa with Ln(3+) and In(3+) yielded complexes of the type [M(H(3)ppa)](3+)(M = La(3+)-Yb(3+)) but with Ga(3+) a neutral complex [Ga(ppa)].3H(2)O was obtained. The formation of an encapsulated 1 : 1 complex is supported by elemental analysis and spectroscopic evidence. PMID- 16357959 TI - New members of the [Ru(diimine)(CN)(4)](2-) family: structural, electrochemical and photophysical properties. AB - A series of complexes of the type K(2)[Ru(NN)(CN)(4)] has been prepared, in which NN is a diimine ligand, and were investigated for both their structural and photophysical properties. The ligands used (and the abbreviations for the resulting complexes) are 3-(2-pyridyl)pyrazole (Ru-pypz), 2,2'-bipyrimidine (Ru bpym), 5,5'-dimethyl-2,2'-bipyridine (Ru-dmb), 1-ethyl-2-(2-pyridyl)benzimidazole (Ru-pbe), bidentate 2,2':6',2'''-terpyridine (Ru-tpy). The known complexes with = 2,2'-bipyridine (Ru-bpy) and 1,10-phenathroline (Ru-phen) were also included in this work. A series of crystallographic studies showed that the [Ru(NN)(CN)(4)](2 ) complex anions form a range of elaborate coordination networks when crystallised with either K(+) or Ln(3+) cations. The K(+) salts are characterised by a combination of near-linear Ru-CN-K bridges, with the cyanides coordinating to K(+) in the usual 'end-on' mode, and unusual side-on pi-type coordination of cyanide ligands to K(+) ions. With Ln(3+) cations in contrast only Ru-CN-Ln near linear bridges occurred, affording 1-dimensional helical or diamondoid chains, and 2-dimensional sheets constituted from linked metallamacrocyclic rings. All of the K(2)[Ru(CN)(4)] complexes show a reversible Ru(II)/Ru(III) couple (ca.+0.9 V vs. Ag/AgCl in water), the exception being Ru-tpy whose oxidation is completely irreversible. Luminescence studies in water showed the presence of (3)MLCT-based emission in all cases apart from Ru-bpym with lifetimes of tens/hundreds of nanoseconds. Time-resolved infrared studies showed that in the (3)MLCT excited state the principal C-N stretching vibration shifts to positive energy by ca. 50 cm(-1) as a consequence of the transient oxidation of the metal centre to Ru(III) and the reduction in back-bonding to the cyanide ligands; measurement of transient decay rates allowed measurements of (3)MLCT lifetimes for those complexes which could not be characterised by luminescence spectroscopy. A few complexes were also examined in different solvents (MeCN, dmf) and showed much weaker emission and shorter excited-state lifetimes in these solvents compared to water. PMID- 16357960 TI - Synthesis of mononuclear and dinuclear ruthenium(II) tris(heteroleptic) complexes via photosubstitution in bis(carbonyl) precursors. AB - A novel, and quite general, approach for the preparation of tris(heteroleptic) ruthenium(II) complexes is reported. Using this method, which is based on photosubstitution of carbonyl ligands in precursors such as [Ru(bpy)(CO)(2)Cl(2)] and [Ru(bpy)(Me(2)bpy)(CO)(2)](PF(6))(2), mononuclear and dinuclear Ru(II) tris(heteroleptic) polypyridyl complexes containing the bridging ligands 3,5 bis(pyridin-2-yl)-1,2,4-triazole (Hbpt) and 3,5-bis(pyrazin-2-yl)-1,2,4-triazole (Hbpzt) have been prepared. The complexes obtained were purified by column chromatography and characterized by HPLC, mass spectrometry, 1H NMR, absorption and emission spectroscopy and by electrochemical methods. The X-ray structures of the compounds [Ru(bpy)(Me(2)bpy)(bpt)](PF(6))x0.5C(4)H(10)O [1x0.5C(4)H(10)O], [Ru(bpy)(Me(2)bpy)(bpzt)](PF(6))xH(2)O (2xH(2)O) and [Ru(bpy)(Me(2)bpy)(CH(3)CN)(2)](PF(6))(2)xC(4)H(10)O (6xC(4)H(10)O) are reported. The synthesis and characterisation of the dinuclear analogues of 1 and 2, [{Ru(bpy)(Me(2)bpy)}(2)bpt](PF(6))(3)x2H(2)O (3) and [{Ru(bpy)(Me(2)bpy)}(2)bpzt](PF(6))(3) (4), are also described. PMID- 16357961 TI - Ligand effect on the kinetics of hydroperoxochromium(III)-oxochromium(V) transformation and the lifetime of chromium(V). AB - A macrocyclic superoxochromium complex L(2)(H(2)O)CrOO(2+)(L(2)=meso-Me(6) [14]aneN(4)) is generated from L(2)Cr(H(2)O)(2)(2+) and O(2) with k(on)=(2.80 +/- 0.07)x 10(7) M(-1) s(-1). One-electron reduction of L(2)(H(2)O)CrOO(2+) produces a transient hydroperoxo complex that readily undergoes intramolecular conversion to L(2)Cr(v), k(1)= 1.00 +/- 0.01 s(-1) in acidic aqueous solutions, and 0.273 +/ 0.010 s(-1) at pH >7, with an apparent pK(a) of 5.9. The decay of L(2)Cr(v) in the pH range 1.3-6.2 obeys the rate law, -d[L(2)Cr(v)]/dt= (0.0080 (+/- 0.0049)+ 8.19 (+/- 0.13)[H(+)])[L(2)Cr(v)]. Both the kinetics of formation and lifetime of L(2)Cr(v) are significantly different from those for the closely related [14]aneN(4) complex. The X-ray structure of the parent Cr(iii) complex, [L(2)Cr(H(2)O)(2)](ClO(4))(3).4H(2)O, shows that the macrocyclic ligand adopts the most stable, "two up-two down" configuration around the nitrogens. PMID- 16357962 TI - The reactivity of gallium-(I), -(II) and -(III) heterocycles towards Group 15 substrates: attempts to prepare gallium-terminal pnictinidene complexes. AB - The reactivity of a series of Ga(I), Ga(II) and Ga(III) heterocyclic compounds towards a number of Group 15 substrates has been investigated with a view to prepare examples of gallium-terminal pnictinidene complexes. Although no examples of such complexes were isolated, a number of novel complexes have been prepared. The reactions of the gallium(I) N-heterocyclic carbene analogue, [K(tmeda)][:Ga{[N(Ar)C(H)](2)}] (Ar = 2,6-diisopropylphenyl) with cyclo-(PPh)(5) and PhN[double bond, length as m-dash]NPh led to the unusual anionic spirocyclic complexes, [{kappa(2)P,P'-(PhP)(4)}Ga{[N(Ar)C(H)](2)}](-) and [{kappa(2)N,C PhNN(H)(C(6)H(4))}Ga{[N(Ar)C(H)](2)}](-), via formal reductions of the Group 15 substrate. The reaction of the digallane(4), [Ga{[N(Ar)C(H)](2)}](2), with (Me(3)Si)N(3) afforded the paramagnetic, dimeric imido-gallane complex, [{[N(Ar)C(H) ](2)}Ga{mu-N(SiMe(3))}](2), via a Ga-Ga bond insertion process. In addition, the new gallium(III) phosphide, [GaI{P(H)Mes*}{[N(Ar)C(H)](2) }], Mes* = C(6)H(2)Bu(t)(3)-2,4,6; was prepared and treated with diazabicycloundecane (DBU) to give [Ga(DBU){P(H)Mes*}{[N(Ar)C(H)](2)}], presumably via a gallium terminal phosphinidene intermediate, [Ga{[double bond, length as m dash]PMes*}{[N(Ar)C(H)](2) }]. The possible mechanisms of all reactions are discussed, all new complexes have been crystallographically characterised and all paramagnetic complexes have been studied by ENDOR and/or EPR spectroscopy. PMID- 16357963 TI - Hydride transfer reactivity of tetrakis(trimethylphosphine)(hydrido)(nitrosyl)molybdenum(0). AB - The tetrakis(trimethylphosphine) molybdenum nitrosyl hydrido complex trans Mo(PMe(3))(4)(H)(NO) (2) and the related deuteride complex trans Mo(PMe(3))(4)(D)(NO) (2a) were prepared from trans-Mo(PMe(3))(4)(Cl)(NO) (1). From (2)H T(1 min) measurements and solid-state (2)H NMR the bond ionicities of 2a could be determined and were found to be 80.0% and 75.3%, respectively, indicating a very polar Mo--D bond. The enhanced hydridicity of 2 is reflected in its very high propensity to undergo hydride transfer reactions. 2 was thus reacted with acetone, acetophenone, and benzophenone to afford the corresponding alkoxide complexes trans-Mo(NO)(PMe(3))(4)(OCHR'R'') (R' = R'' = Me (3); R' = Me, R'' = Ph (4); R' = R'' = Ph (5)). The reaction of 2 with CO(2) led to the formation of the formato-O-complex Mo(NO)(OCHO)(PMe(3))(4) (6). The reaction of with HOSO(2)CF(3) produced the anion coordinated complex Mo(NO)(PMe(3))(4)(OSO(2)CF(3)) (7), and the reaction with [H(Et(2)O)(2)][BAr(F)(4)] with an excess of PMe(3) produced the pentakis(trimethylphosphine) coordinated compound [Mo(NO)(PMe(3))(5)][BAr(F)(4)] (8). Imine insertions into the Mo-H bond of 2 were also accomplished. PhCH[double bond, length as m-dash]NPh (N-benzylideneaniline) and C(10)H(7)CH=NPh (N-1 naphthylideneaniline) afforded the amido compounds Mo(NO)(PMe(3))(4)[NR'(CH(2)R'')] (R' = R'' = Ph (9), R' = Ph, R'' = naphthyl (11)). 9 could not be obtained in pure form, however, its structure was assigned by spectroscopic means. At room temperature 11 reacted further to lose one PMe(3) forming 12 (Mo(NO)PMe(3))(3)[N(Ph)CH(2)C(10)H(6))]) with agostic stabilization. In a subsequent step oxidative addition of the agostic naphthyl C-H bond to the molybdenum centre occurred. Then hydrogen migration took place giving the chelate amine complex Mo(NO)(PMe(3))(3)[NH(Ph)(CH(2)C(10)H(6))] (15). The insertion reaction of 2 with C(10)H(7)N=CHPh led to formation of the agostic compound Mo(NO)(PMe(3))(3)[N(CH(2)Ph)(C(10)H(7))] (10). Based on the knowledge of facile formation of agostic compounds the catalytic hydrogenation of C(10)H(7)N=CHPh and PhN=CHC(10)H(7) with 2 (5 mol%) was tested. The best conversion rates were obtained in the presence of an excess of PMe(3), which were 18.4% and 100% for C(10)H(7)N=CHPh and PhN=CHC(10)H(7), respectively. PMID- 16357964 TI - Tetrametallic clusters (Ir(2)Rh(2)) through an ancillary ortho-carborane-1,2 dichalcogenolato ligands. AB - The tetrametallic cluster complexes {Cp*Ir[E(2)C(2)(B(10)H(9))]}Rh(2)(cod){Cp*Ir[E(2)C(2) (B(10)H(10))]} (E = S; Se) have been synthesized by reactions of the 16-electron half-sandwich iridium complexes [Cp*Ir{E(2)C(2)(B(10)H(10))}] [Cp* = eta(5)-C(5)Me(5), E = S, Se] with [Rh(cod)(micro-OEt)(2)] at room temperature in toluene solution. In the solid state, this tetrametallic cluster exhibits an irregular nearly planar metal skeleton with the two carborane dichalcogenolato ligands bridging the four metal centers from both sides of the tetrametallic plane. Even though all metal atoms coordinate bridging chalcogen atoms, they show different electronic and coordination environments. The molecular structures of and have been determined by X-ray crystallography. PMID- 16357965 TI - The synthesis, characterisation and reactivity of 2 phosphanylethylcyclopentadienyl complexes of cobalt, rhodium and iridium. AB - 2-Phosphanylethylcyclopentadienyl lithium compounds, Li[C(5)R'(4)(CH(2))(2)PR(2)] (R = Et, R' = H or Me, R = Ph, R' = Me), have been prepared from the reaction of spirohydrocarbons C(5)R'(4)(C(2)H(4)) with LiPR(2). C(5)Et(4)HSiMe(2)CH(2)PMe(2), was prepared from reaction of Li[C(5)Et(4)] with Me(2)SiCl(2) followed by Me(2)PCH(2)Li. The lithium salts were reacted with [RhCl(CO)(2)](2), [IrCl(CO)(3)] or [Co(2)(CO)(8)] to give [M(C(5)R'(4)(CH(2))(2)PR(2))(CO)] (M = Rh, R = Et, R' = H or Me, R = Ph, R' = Me; M = Ir or Co, R = Et, R' = Me), which have been fully characterised, in many cases crystallographically as monomers with coordination of the phosphorus atom and the cyclopentadienyl ring. The values of nu(CO) for these complexes are usually lower than those for the analogous complexes without the bridge between the cyclopentadienyl ring and the phosphine, the exception being [Rh(Cp'(CH(2))(2)PEt(2))(CO)] (Cp' = C(5)Me(4)), the most electron rich of the complexes. [Rh(C(5)Et(4)SiMe(2)CH(2)PMe(2))(CO)] may be a dimer. [Co(2)(CO)(8)] reacts with C(5)H(5)(CH(2))(2)PEt(2) or C(5)Et(4)HSiMe(2)CH(2)PMe(2) (L) to give binuclear complexes of the form [Co(2)(CO)(6)L(2)] with almost linear PCoCoP skeletons. [Rh(Cp'(CH(2))(2)PEt(2))(CO)] and [Rh(Cp'(CH(2))(2)PPh(2))(CO)] are active for methanol carbonylation at 150 degrees C and 27 bar CO, with the rate using [Rh(Cp'(CH(2))(2)PPh(2))(CO)] (0.81 mol dm(-3) h(-1)) being higher than that for [RhI(2)(CO)(2)](-) (0.64 mol dm(-3) h(-1)). The most electron rich complex, [Rh(Cp'(CH(2))(2)PEt(2))(CO)] (0.38 mol dm(-3) h(-1)) gave a comparable rate to [Cp*Rh(PEt(3))(CO)] (0.30 mol dm(-3) h(-1)), which was unstable towards oxidation of the phosphine. [Rh(Cp'(CH(2))(2)PEt(2))I(2)], which is inactive for methanol carbonylation, was isolated after the methanol carbonylation reaction using [Rh(Cp'(CH(2))(2)PEt(2))(CO)]. Neither of [M(Cp'(CH(2))(2)PEt(2))(CO)] (M = Co or Ir) was active for methanol carbonylation under these conditions, nor under many other conditions investigated, except that [Ir(Cp'(CH(2))(2)PEt(2))(CO)] showed some activity at higher temperature (190 degrees C), probably as a result of degradation to [IrI(2)(CO)(2)](-). [M(Cp'(CH(2))(2)PEt(2))(CO)] react with MeI to give [M(Cp'(CH(2))(2)PEt(2))(C(O)Me)I] (M = Co or Rh) or [Ir(Cp'(CH(2))(2)PEt(2))Me(CO)]I. The rates of oxidative addition of MeI to [Rh(C(5)H(4)(CH(2))(2)PEt(2))(CO)] and [Rh(Cp'(CH(2))(2)PPh(2))(CO)] are 62 and 1770 times faster than to [Cp*Rh(CO)(2)]. Methyl migration is slower, however. High pressure NMR studies show that [Co(Cp'(CH(2))(2)PEt(2))(CO)] and [Cp*Rh(PEt(3))(CO)] are unstable towards phosphine oxidation and/or quaternisation under methanol carbonylation conditions, but that [Rh(Cp'(CH(2))(2)PEt(2))(CO)] does not exhibit phosphine degradation, eventually producing inactive [Rh(Cp'(CH(2))(2)PEt(2))I(2)] at least under conditions of poor gas mixing. The observation of [Rh(Cp'(CH(2))(2)PEt(2))(C(O)Me)I] under methanol carbonylation conditions suggests that the rhodium centre has become so electron rich that reductive elimination of ethanoyl iodide has become rate determining for methanol carbonylation. In addition to the high electron density at rhodium. PMID- 16357966 TI - Controlled formation and topologies of thiophenolate-based macrocycles: rings, cylinders and bowls. AB - The Schiff-base condensations of 1,3-diaminopropane with a protected thiophenol dialdehyde in the presence of Ni(2+), Pd(2+) or Zn(2+) can be controlled to yield either mononuclear acyclic, or 2 + 2 and 4 + 4 macrocyclic complexes by the choice of both metal cation and counteranion. The Ni(2+) complex of the 2 + 2 macrocycle contains two square-planar nickel ions and shows an arrangement similar to one observed previously: the mu-S atoms of the thiophenolate groups are pyramidal and lie on the same side of the plane defined by the four N atoms of the macrocycle to give a V-shaped molecule. By contrast, the Zn(2+) complex of the 2 + 2 macrocycle undergoes oligomerization to yield a bowl-shaped hexanuclear complex that includes a mu(3)-carbonate anion. Essential for this topology is the presence of three mu(3)-S-thiophenolato groups that link the three macrocyclic units to form a Zn(3)S(3) ring that seals the bottom part of the bowl. In this arrangement, one of the pyramidal mu(3)-S atoms in each dinuclear Zn(2+) complex is inverted relative to the arrangement observed for the dinickel complexes. Molecular modelling suggests that inversion about the mu-S atoms of the 2 + 2 macrocyclic complexes is readily accessible at room temperature and that the contrasting arrangements observed for the Ni(2+) and Zn(2+) complexes are those energetically most favourable for the respective metal ions. Rare 4 + 4 macrocyclic complexes are isolated as neutral dinuclear complexes for Ni(2+) and Pd(2+) and as a tetranuclear complex cation for Zn(2+). The topologies of these systems contrast significantly: those with two square-planar Ni(2+) or Pd(2+) ions form extended rings, while that with Zn(2+) forms a sulfur-lined cylinder which hosts acetonitrile molecules in the crystalline state. Reaction conditions can also be optimised to produce 2 + 1 acyclic ligands as their mononuclear Ni(2+) and Pd(2+) complexes, providing potentially useful building blocks for production of more complicated macrocyclic and supramolecular systems. PMID- 16357967 TI - Mixed hydrazido amido/imido complexes of tantalum, hafnium and zirconium: potential precursors for metal nitride MOCVD. AB - The coordination chemistry of the hydrazine derivatives dimethylhydrazine (Hdmh) and N-trimethylsilyl-N'N'-dimethylhydrazine (Htdmh) at Ta, Zr and Hf was investigated aiming at volatile mixed ligand all-nitrogen coordinated compounds. The hydrazido ligands were introduced either by salt metathesis employing the Li salts of the hydrazines and the tetrachlorides MCl(4) (M = Zr, Hf) or by amine substitution using M(NR(2))(4) (R = Me, Et) and [(t-BuN)Ta(NR(2))(3)]. The new complexes were fully characterised including (1)H/(13)C NMR, mass spectrometry and a study of their thermal behaviour. The crystal structures of [ZrCl(tdmh)(3)] and the all-nitrogen coordinated complex [Ta(N-t-Bu)(NMe(2))(2)(tdmh)] are discussed as well as the structure of the by-product [Li(tdmh)(py)](2). Preliminary MOCVD experiments of the liquid compound [Ta(NEt(2))(2)(N-t Bu)(tdmh)] were performed and the deposited TaN(Si) films were analysed by RBS and SEM. PMID- 16357968 TI - Sodium compounds of the benzophenone dianion (diphenyloxidomethanide). AB - The benzophenone dianion [diphenyloxidomethanide, (Ph(2)CO)(2-)], which occurs in the well known deeply violet sodium/benzophenone tetrahydrofuran solutions, was crystallised with sodium cations in form of the two polymeric chain compounds [Na(2)(Ph(2)CO)(tetraglyme)](infinity) and [Na(2)(Ph(2)CO)(thf)(2)](infinity). It was found to aggregate with its conjugated acid, the alcoholate (Ph(2)CHO)(-), around a central unit of sodium hydroxide, resulting in the mixed cage compound [Na(13)(Ph(2)CO)(4)(Ph(2)CHO)(4)(OH)(mtbe)(4)].mtbe. The structural parameters of the benzophenone dianion indicate that a considerable amount of its negative charge is located within the phenyl rings, rather than on the formally anionic benzylic carbon atom. The topological analysis of the electron density of the monomeric model structure [Na(2)(Ph(2)CO)] reveals an even positive charge for this particular atom, hence (Ph(2)CO)(2-) is, despite its usual representation, not a vicinal dianion. PMID- 16357969 TI - Coordination chemistry of phosphanyl amino acids: solid state and solution structures of neutral and cationic rhodium complexes. AB - Copper phosphide or arsenide complexes, [Cu(EPh(2))(neo)] (E = P, As, neo = 2,9 dimethyl-1,10-phenanthroline; trivial name: neocuprine) react selectively with the N-protected brominated serine derivatives, 2-(S)-(alkoxycarbonylamino)-3 bromomethylpropionates ((ROCO)SerBr, : R = PhCH(2), : tBu, : Me) to give the corresponding phosphanylated or arsanylated amino acids, (ROCO)SerPhos (: Phos = PPh(2)) and (Z)SerArs (Ars = AsPh(2), Z = PhCH(2)OCO). The dipeptide (Z)AlaSerPhos was likewise prepared. The phosphanes , and the arsane reacted cleanly with [Rh(2)(micro-Cl)(2)(cod)(2)] to give the rhodium(I) complexes [RhCl(cod)((Z)SerPhos)] , [RhCl(cod)((Boc)SerPhos)] (Boc = tBuOCO), [RhCl(cod)((Z)AlaSerPhos)] , and [RhCl(cod)((Z)SerArs)] which were characterized by X-ray diffraction studies. A common structural feature is an intramolecular (N)H[dot dot dot]Cl(Rh)-hydrogen bridge which according to NMR investigations remains intact in solution. The abstraction of chloride from the coordination sphere of Rh(I) in or has a profound structural impact. While in and , the ligands bind in a monodentate fashion, via the phosphorus atom only, they serve as bidentate ligands via the phosphorus centre and the peptidic C=O group in [Rh(cod)(kappa(2)-(Z)SerPhos)]PF(6) and [Rh(cod)(kappa(2)-(Z)AlaSerPhos)]PF(6). This causes also the amino acid residue structures to change from alpha-helix type in and to a beta-sheet type in both. Addition of chloride to and fully re establishes the structures of both. The complexes [RhCl(cod)((Z)SerPhos)] and [RhCl(cod)((Boc)SerPhos)] show good activities in homogeneously catalyzed hydrogenations of olefins while the dipeptide complex is less active. Phosphane addition to greatly diminishes the catalytic activity. The cationic complex [Rh(cod)(kappa(2)-(Z)AlaSerPhos)]PF(6) shows low activity which, however, is greatly increased by addition of one equivalent of phosphane. PMID- 16357970 TI - Solution-phase template approach for the synthesis of Cu(2)S nanoribbons. AB - In this paper, we have developed a solution-phase template approach to synthesize Cu(2)S nanoribbons for the first time. Bi(2)S(3) nanoribbons act as both template and reactant when treated with small CuCl particles, generating Cu(2)S nanoribbons with the assistance of the solvent ethanol. Nanoribbons with different compositions of Bi(2)S(3) and Cu(2)S also could be obtained through controlling the reaction time. This kind of template method is expected to be a general template approach due to its slow reaction rate and simplicity. PMID- 16357971 TI - Coordination chemistry of 1,4-bis-carboxymethylcyclam, H(2)(1,4-bcc). AB - Zinc metal reduction of the cobalt(III) complex [Co(1,4-bcc)](+) (1,4-bcc = 1,4 bis-carboxymethylcyclam) produces the corresponding cobalt(II) complex which crystallises as the coordination polymer {[Co(1,4-bcc)]ZnCl(2)}(n). A method has been developed for removal of the cobalt(III) ion from [Co(1,4-bcc)](+) and isolation of the free ligand as its hydrochloride salt, H(2)(1,4-bcc).4HCl. This has been used for the preparation of new metal complexes, and the syntheses and characterisation of the copper(ii), nickel(ii), zinc(ii) and chromium(iii) complexes containing the 1,4-bcc ligand are described. X-Ray crystal structures of {[Co(1,4-bcc)]ZnCl(2)}(n).2.5H(2)O, {[Cu(1,4-bcc)]CuCl(2)}(n).0.25MeOH.H(2)O and [Cu(1,4-bcc)H]ClO(4) show the complexes to have the trans(O) geometry of the 1,4-bcc ligand, while the structure of [Cr(1,4-bcc)H(0.5)](ClO(4))(1.5).EtOH exhibits the cis(O) configuration. PMID- 16357972 TI - Catalytic alcohol oxidation by an unsymmetrical 5-coordinate copper complex: electronic structure and mechanism. AB - Density functional theory reveals the detailed mechanism of alcohol oxidation by a model copper complex, Cu(II)L, L = cis-1-(3',5'-dimethoxy-benzylideneamino)-3,5 [2-hydroxy-(3',5'-di-tert-butyl)benzylideneimino]cyclohexane. Despite the obvious structural and functional parallels between the title compound and the enzyme galactose oxidase, the details of the catalytic pathway are fundamentally different. In the enzyme, coordination of the substrate produces an active form containing a Cu(II) centre and a tyrosyl radical, the latter being responsible for the abstraction of hydrogen from the substrate. In the model system, in marked contrast, the active form contains a Cu(II) centre, but the ligand radical character is localised on the substrate (alcoholate) oxygen, rather than the phenolate ligand. The result is a significantly higher barrier to hydrogen-atom abstraction compared to the enzyme itself. The origin of these significant differences is traced to the rigid nature of the pentadentate ligand, which resists changes in coordination number during the catalytic cycle. PMID- 16357973 TI - Electronic structures of trans-dioxometal complexes. AB - We have employed computational methods based on density functional theory to elucidate the effects of equatorial ligands on the electronic structures of trans dioxometal complexes. In complexes with amine (sigma-only) equatorial donors, the (1)A(1 g)(b(2 g))(2)-->(1)E(g)(b(2 g))(1)(e(g))(1) excitation energy increases with metal oxidation state: Mo(IV) < Tc(V) < Ru(vi) and W(IV) < Re(V) < Os(VI). Increasing transition energies are attributed to enhanced oxometal pi-donor interactions in the higher valent central metals. But in complexes with cyanide equatorial donors, the (1)A(1 g)(b(2 g))(2)-->(1)E(g)(b(2 g))(1)(e(g))(1) energy remains roughly independent of metal oxidation state, likely owing to the compensating increased pi-donation from the pi(CN) orbitals to the metal d(xy) orbitals as the oxidation state of the metal increases. PMID- 16357974 TI - (N-Benzyl-bis-N',N''-salicylidene)-cis-1,3,5-triaminocyclohexane copper(II): a novel catalyst for the aerobic oxidation of benzyl alcohol. AB - Reaction of Cu(BF(4))(2).6H(2)O with the N(3)O(2) donor ligand H(2)L (where H(2)L = N-benzyl-N',N''-di-tert-butyl-disalicyl-triaminocyclohexane) results in the formation of a novel Cu(II)L complex, 1. X-Ray crystallography of it shows the Cu(II) centre coordinated by two phenolate oxygens and two imine nitrogens in a distorted square plane with an elongated bond to the amine nitrogen (2.512 A) in the axial position. EPR spectroscopy gives g values of g(1) = 2.277, g(2) = 2.100, g(3) = 2.025, and A(1) = 15.6 mT which are consistent with the distorted square pyramidal coordination environment determined from the X-ray structure. UV/visible and electrochemical analysis of shows that it undergoes two reversible processes assigned to the successive oxidation of the phenolate oxygens to phenoxyl radicals, the first at E((1/2)) = 0.89 V (DeltaE = 81 mV, vs. Ag/AgCl) and the second at E((1/2)) = 1.13V (DeltaE = 84 mV, vs. Ag/AgCl). Chemical oxidation results in the formation of a species, assigned as [1](+)(.) which is EPR silent due to antiferromagnetic coupling between the Cu(II) centre and the bound phenoxyl radical. The oxidised species catalyses the oxidation of benzyl alcohol to benzaldehyde. PMID- 16357975 TI - Ligand dependence of pi-complex character in disilene-palladium complexes. AB - The synthesis and structures of new 16-electron disilene palladium complexes with 2,6-dimethylphenyl isocyanide and phenyldimethylphosphine ligands [L(1)L(2)Pd{(t BuMe(2)Si)(2)Si=Si(SiMe(2)Bu-t)(2)}, where L(1) = L(2) = PhMe(2)P; L(1) = (cyclohexyl)(3)P, L(2) = 2,6-dimethylphenyl isocyanide; L(1) = L(2) = 2,6 dimethylphenyl isocyanide] are described. Comparison of the X-ray structural parameters around the disilene moiety among these complexes and related bis(trimethylphosphine)(disilene)palladium and 14-electron (tricyclohexylphosphine)(disilene)palladium revealed that the pi-complex character is sensitive to the residual ligands and increases with decreasing the strength of sigma-donation from the ligands. PMID- 16357976 TI - Thioester hydrolysis reactivity of zinc hydroxide complexes: investigating reactivity relevant to glyoxalase II enzymes. AB - A recently reported binuclear zinc hydroxide complex [(L(1)Zn(2))(mu OH)](ClO(4))(2) (, L(1) = 2,6-bis[(bis(2-pyridylmethyl)amino)methyl]-4 methylphenolate monoanion) containing a single bridging hydroxide was examined for thioester hydrolysis reactivity. Treatment of it with hydroxyphenylthioacetic acid S-methyl ester in dry CD(3)CN results in no reaction after approximately 65 h at 45(1) degrees C. Binuclear zinc hydroxide complexes of the N-methyl-N-((6 neopentylamino-2-pyridyl)methyl)-N-((2-pyridyl)methyl)amine (L(2)) and N-methyl-N ((6-neopentylamino-2-pyridyl)methyl)-N-((2-pyridyl)ethyl)amine (L(3)) chelate ligands were prepared by treatment of each ligand with molar equivalent amounts of Zn(ClO(4))(2).6H(2)O and KOH in methanol. These complexes, [(L(2)Zn)(2)(mu OH)(2)](ClO(4))(2) and [(L(3)Zn)(2)(mu-OH)(2)](ClO(4))(2) (), which have been structurally characterized by X-ray crystallography, behave as 1 : 1 electrolytes in acetonitrile, indicating that the binuclear cations dissociate into monomeric zinc hydroxide species in solution. Treatment of them with one equivalent of hydroxyphenylthioacetic acid S-methyl ester per zinc center in acetonitrile results in the formation of a zinc alpha-hydroxycarboxylate complex, [(L(2))Zn(O(2)CCH(OH)Ph)]ClO(4).1.5H(2)O or [(L(3))Zn(O(2)CCH(OH)Ph)]ClO(4).1.5H(2)O, and CH(3)SH. These reactions, to our knowledge, are the first reported examples of thioester hydrolysis mediated by zinc hydroxide complexes. The results of this study suggest that a terminal Zn-OH moiety may be required for hydrolysis reactivity with a thioester substrate. PMID- 16357977 TI - Bisoxazolines with one and two sidearms: stereodirecting ligands for copper catalysed asymmetric allylic oxidations of alkenes. AB - A series of sidearm functionalized bisoxazoline ligands has been synthesized by reaction of the monolithiated methyl{bis(oxazolinyl)}methane with the appropriate electrophiles, and tested in the copper catalyzed asymmetric allylic oxidation of cyclohexene ("Kharasch-Sosnovski" reaction). The observed enantioselectivities were higher (up to 85% ee) than for the unfunctionalized bisoxazoline ("BOX") derivatives (ca. 60% ee). Regardless of the functional groups incorporated into the sidearm unit, the ee's obtained for the different derivatives were essentially indistinguishable. This implies that the sidearms do not interfere directly in this reaction and only play an indirect role by virtue of their steric demand. Three of the copper complexes have been characterized by X-ray diffraction, establishing a distorted octahedral coordination geometry around the copper atom in all three cases. In the elongated distorted CuN(2)O(4) octahedra, the two nitrogen atoms of the oxazolines and one oxygen atom of each acetate ligand occupy the 'equatorial' positions whereas the sidearms do not interact with the metal centres. PMID- 16357978 TI - Olefination and group transfer reactions of an electron deficient tantalum methylidene complex. AB - The reactivity of an electronically unsaturated tantalum methylidene complex [TolC(NSiMe(3))(2)](2)Ta(CH(2))CH(3) supported by [TolC(NSiMe(3))(2)] amidinate ligands is described. Electrophilic addition and olefination reactions of the Ta=CH(2) functionality are reported. Alkylidene participates in group-transfer reactions not observed in sterically similar, but electronically saturated, analogues. Reactions with substrates containing unsaturated C-X (X = C, N, O) bonds yield [Ta]=X compounds and vinylated organic products; carbon-sulfur cleavage reactions to produce tantalum thioformaldehyde and tantalum sulfido complexes. PMID- 16357979 TI - Chemistry of palladium phosphinite (PPh(2)(OR)) and phosphonite (P(OPh)(2)(OH)) complexes: catalytic activity in methoxycarbonylation and Heck coupling reactions. AB - The new phosphinite and phosphonite complexes (1-8) are very efficient catalysts for the methoxycarbonylation of iodobenzene and Heck cross-coupling of bromobenzene with butyl acrylate. High catalytic activity of these complexes can be explained by their in situ transformations during the reaction, stimulated by the presence of water, acid (HCl) or base (NEt(3)). Hydrolysis of phosphinite palladium complexes of the form trans-PdCl(2)[PPh(2)(OR)](2) (R = C(6)F(5), 2, (t)Bu 3, or O-menthyl 4) results in the formation of the dimeric complex [mu ClPd(PPh(2)OH)(PPh(2)O)](2) 5, which is deprotonated by NEt(3), producing a polymeric complex of formula [Pd(P(O)PPh(2))(2)](n) 8. The reverse reaction, protonolysis of 8 with HCl, leads back to 5 and the monomeric complex 5a. The phosphinite complex PdCl(2)[PPh(2)(OBu)](2)1 with a more lipophilic ligand, PPh(2)(OBu), does not undergo hydrolysis under the same conditions. In the reaction of PdCl(2)(cod) with P(OPh)(2)(OH), the new dimer [mu ClPd(P(OPh)(2)OH)(P(OPh)(2)O)](2) 6 was obtained, whereas reaction of Pd(OAc)(2) with P(OPh)(2)(OH) leads to the polymeric complex [Pd[P(O)(OPh)(2)](2)](n) 7. Protonolysis of 7 with HCl results in the formation of 6. PMID- 16357980 TI - 1, 2-, 1, 3- and 1, 4-Cyclohexanedicarboxylates of Cd and Mn with chain and layered structures. AB - A systematic study has been carried out on the three isomeric cyclohexanedicarboxylates (CHDCs) formed by cadmium and manganese with the three isomeric dicarboxylic acids, in the presence or absence of amines. The CHDCs have been prepared under hydrothermal conditions and their structures established by X ray crystallography. We have been able to isolate two-dimensional layered structures of 1,2-, 1,3- and 1,4-cyclohexanedicarboxylates and chain structures of 1,3- and 1,4-cyclohexanedicarboxylates. The infinite metal-oxygen-metal linkages are observed only in the case of the 1,2-dicarboxylate. In all the three isomeric cyclohexanedicarboxylates, the e,e conformation is most favored, although the 1,4-CHDCs often contain rings in both the e,e and the a,e conformations. PMID- 16357981 TI - Single-walled carbon nanotube growth using [Fe(3)(mu(3)-O)(mu O(2)CR)(6)(L)(3)](n+) complexes as catalyst precursors. AB - We present herein the VLS growth of SWNTs from oxo-hexacarboxylate-triron precursors, [Fe(3)O(O(2)CCH(3))(6)(EtOH)(3)] and [Fe(3)O(O(2)CCH(2)OMe)(6)(H(2)O)(3)][FeCl(4)], on spin-on-glass surfaces, using C(2)H(4)/H(2) (750 degrees C) and CH(4)/H(2) (800 and 900 degrees C) growth conditions. The SWNTs have been characterized by AFM, SEM and Raman spectroscopy. The characteristics of the SWNTs are found to be independent of the identity of the precursor complex or the solvent from which it is spin-coated. The as grown SWNTs show a low level of side-wall defects and have an average diameter of 1.2 1.4 nm with a narrow distribution of diameters. At 750 and 800 degrees C the SWNTs are grown with a range of lengths (300 nm-9 microm), but at 900 degrees C only the longer SWNTs are observed (6-8 microm). The yield of SWNTs per unit area of catalyst nanoparticle decreases with the growth temperature. We have demonstrated that spin coating of molecular precursors allows for the formation of catalyst nanoparticles suitable for growth of SWNTs with a high degree of uniformity in the diameter, without the formation of preformed clusters of a set diameter. PMID- 16357982 TI - Copolymerization of cyclohexene oxide and carbon dioxide using (salen)Co(III) complexes: synthesis and characterization of syndiotactic poly(cyclohexene carbonate). AB - Synthetic routes to a series of new (salen-1)CoX (salen-1 = N,N' bis(salicylidene)-1,2-diaminoalkane; X = halide or carboxylate) species are described and the X-ray crystal structures of two (salen-1)CoX (salen- = N,N' bis(3,5-di-tert-butylsalicylidene)-1,2-diaminocyclohexane; X = Cl, I) complexes are presented. (R,R)-(salen-)CoX (X = Cl, Br, I, OAc, pentafluorobenzoate (OBzF(5))) catalysts are active for the copolymerization of cyclohexene oxide (CHO) and CO(2), yielding syndiotactic poly(cyclohexene carbonate) (PCHC), a previously unreported PCHC microstructure. Variation of the salen ligand and reaction conditions, as well as the inclusion of [PPN]Cl ([PPN]Cl = bis(triphenylphosphine)iminium chloride) cocatalysts, has dramatic effects on the polymerization rate and the resultant PCHC tacticity. Catalysts rac-(salen-)CoX (salen- = N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-diaminopropane; X = Br, OBzF(5)) have high activities for CHO/CO(2) copolymerization, yielding syndiotactic PCHCs with up to 81% r-centered tetrads. Using Bernoullian statistical methods, PCHC tetrad and triad sequences were assigned in the (13)C NMR spectra of these polymers in the carbonyl and methylene regions, respectively. PMID- 16357983 TI - Interplay between lead carboxylate and Ti or Zr isopropoxides in solution routes to perovskites: synthesis, molecular structures and reactivity of single source non-oxo Pb-Zr and Pb-Ti carboxylatoalkoxides supported by 2-ethylhexanoate ligands. AB - The reactions between Ti(O(i)Pr)(4) and Zr(2)(O(i)Pr)(8)(HO(i)Pr)(2), respectively, and lead 2-ethylhexanoate Pb(O(2)CC(7)H(15))(2) have been investigated at rt and by heating. The initial mixed-metal species, characterized by single-crystal X-Ray diffraction, were adducts namely Pb(4)Zr(4)(mu O(2)CR')(8)(mu-OR)(6)(mu(3)-OR)(2)(OR)(8)(OHR)(2) and Pb(2)Ti(4)(mu O(2)CR')(4)(mu-OR)(6)(mu(3)-OR)(2)(OR)(8) (R' = CHCH(Et)C(2)H(4)Me, R = (i)Pr) independently of the stoichiometry used. They are the first Pb-Ti and Pb-Zr non oxo carboxylatoalkoxides reported. is also the first Pb-Zr species based on an alkoxide-carboxylate ligand set matching the PbZrO(3) stoichiometry. Both structures are centrosymmetric with six-coordinate transition metals, as required for the perovskite, and are based on triangular M(2)Pb cores (M = Zr, Ti). The lead centers display quite high coordination numbers, six and seven. The thermal and hydrolytic condensation reactions of and were investigated. Heat treatment of and elimination of the volatiles under vacuum afforded Pb(2)Ti(3)(mu(4)-O)(mu(3) O)(mu-O(2)CC(7)H(15))(2)(mu-O(i)Pr)(6)(O(i)Pr)(4) resulting from extrusion of Ti(O(i)Pr)(4) and scrambling of carboxylate ligands. Characterization of the various compounds was achieved by elemental analysis, FT-IR, (1)H and (207)Pb NMR. PMID- 16357984 TI - Phenoxyl radicals: H-bonded and coordinated to Cu(II) and Zn(II). AB - Two pro-ligands ((R)LH) comprised of an o,p-di-tert-butyl-substituted phenol covalently bonded to a benzimidazole ((Bz)LH) or a 4,5-di-p-methoxyphenyl substituted imidazole ((PhOMe)LH), have been structurally characterised. Each possesses an intramolecular O-H[dot dot dot]N hydrogen bond between the phenolic O-H group and an imidazole nitrogen atom and (1)H NMR studies show that this bond is retained in solution. Each (R)LH undergoes an electrochemically reversible, one-electron, oxidation to form the [(R)LH] (+) radical cation that is considered to be stabilised by an intramolecular O...H-N hydrogen bond. The (R)LH pro ligands react with M(BF(4))(2).H(2)O (M = Cu or Zn) in the presence of Et(3)N to form the corresponding [M((R)L)(2)] compound. [Cu((Bz)L)(2)] (), [Cu((PhOMe)L)(2)] (), [Zn((Bz)L)(2)] and [Zn((PhOMe)L)(2)] have been isolated and the structures of .4MeCN, .2MeOH, .2MeCN and .2MeCN determined by X-ray crystallography. In each compound the metal possesses an N(2)O(2)-coordination sphere: in .4MeCN and .2MeOH the {CuN(2)O(2)} centre has a distorted square planar geometry; in .2MeCN and .2MeCN the {ZnN(2)O(2)} centre has a distorted tetrahedral geometry. The X-band EPR spectra of both and , in CH(2)Cl(2)-DMF (9 : 1) solution at 77 K, are consistent with the presence of a Cu(ii) complex having the structure identified by X-ray crystallography. Electrochemical studies have shown that each undergo two, one-electron, oxidations; the potentials of these processes and the UV/vis and EPR properties of the products indicate that each oxidation is ligand-based. The first oxidation produces [M(II)((R)L)((R)L )](+), comprising a M(ii) centre bound to a phenoxide ((R)L) and a phenoxyl radical ((R)L ) ligand; these cations have been generated electrochemically and, for R = PhOMe, chemically by oxidation with Ag[BF(4)]. The second oxidation produces [M(II)((R)L )(2)](2+). The information obtained from these investigations shows that a suitable pro-ligand design allows a relatively inert phenoxyl radical to be generated, stabilised by either a hydrogen bond, as in [(R)LH] (+) (R = Bz or PhOMe), or by coordination to a metal, as in [M(II)((R)L)((R)L )](+) (M = Cu or Zn; R = Bz or PhOMe). Coordination to a metal is more effective than hydrogen bonding in stabilising a phenoxyl radical and Cu(ii) is slightly more effective than Zn(II) in this respect. PMID- 16357985 TI - SPANphos: trans-spanning diphosphines as cis chelating ligands! AB - Several SPANphos ligands based on a spirobichroman backbone, introduced as a putative trans ligand, form compounds of the type [Rh(nbd)(SPANphos)]BF(4) () in which both norbornadiene and SPANphos act as cis chelating ligands. The cyclooctadiene rhodium chloride derivatives form bimetallic complexes. Crystal structures for several of these compounds and free ligands are reported. Semiemperical AM1 and DFT calculations show that spirobichroman can assume several conformations, some of which are suitable for the formation of cis chelating SPANphos. All calculations on SPANphos complexes of PdCl(2), PtCl(2) and Rh(CO)Cl show that the trans complex is more stable by 4-10 kcal mol(-1). The cis conformation is enforced by the cis chelating norbornadiene ligand. PMID- 16357986 TI - Synthesis, characterization and reactivity of tetranuclear ruthenium hydrido clusters containing chiral phosphine ligands. AB - The chiral clusters [H(4)Ru(4)(CO)(12-n)(L)(n)] (n = 1, 2; L = NMDPP), 1,1 [H(4)Ru(4)(CO)(10)(L-L)] (L-L = DUPHOS, DIPAMP), 1,2-[H(4)Ru(4)(CO)(10)(DIOP)] and [{H(4)Ru(4)(CO)(10)(DIOP)}(2)] have been synthesized by derivatizing the parent carbonyl cluster [H(4)Ru(4)(CO)(12)] with the appropriate mono- or didentate chiral phosphine ligand. The phosphine-substituted clusters were found to be able to catalyze the (asymmetric) hydrogenation of tiglic acid albeit with relatively low selectivity (enantiomeric excesses varying from 0 to 23%). It was found that the stability of the chiral ruthenium hydride clusters and the product distribution obtained in the catalytic reactions are dependent on the nature of the chiral phosphine. The crystal structures of [H(4)Ru(4)(CO)(12-n)(L)(n)] (n = 1, 2; L = NMDPP), 1,1-[H(4)Ru(4)(CO)(10)(L-L)] (L-L = DUPHOS, O-DUPHOS (partially oxygenated ligand), DIPAMP), 1,2-[H(4)Ru(4)(CO)(10)(DIOP)] and [{H(4)Ru(4)(CO)(10)(DIOP)}(2)] are presented. PMID- 16357988 TI - Artificial ribonucleases. AB - Mimicking the action of enzymes by simpler and more robust man-made catalysts has long inspired bioorganic chemists. During the past decade, mimics for RNA cleaving enzymes, ribonucleases, or, more precisely, mimics of ribozymes that cleave RNA in sequence-selective rather than base-selective manner, have received special attention. These artificial ribonucleases are typically oligonucleotides (or their structural analogs) that bear a catalytically active conjugate group and catalyze sequence-selective hydrolysis of RNA phosphodiester bonds. PMID- 16357989 TI - Electrophilic fluorination of organosilanes. AB - The fluorination of organosilanes with the silyl groups directly attached or adjacent to an aryl or alkenyl group has been only very recently examined despite the fact that the corresponding fluorinated products are synthetically useful building blocks. In these reactions, the silyl group enhances the reactivity of the pi-nucleophile and controls the sense of regiochemistry upon addition of the electrophilic source of fluorine. These reactions take advantage of the beta effect of the silicon-carbon bond and recent results from the literature revealed that this chemistry allows for the preparation of a variety of novel fluorinated building blocks including enantio-enriched derivatives. PMID- 16357990 TI - Aromatic annulation strategy for naphthalenes fused at 1,2- and 3,4-positions with two heterocycles. AB - An efficient regioselective naphthoannulation strategy able to fuse a newly formed naphthalene ring at its 1,2- and 3,4- positions to two different heterocycles has been developed. PMID- 16357991 TI - Mitsunobu dehydration of N-Boc neomycin B. AB - Reaction of hexa-N-Boc neomycin B with TPP and DIAD in toluene results in the formation of an epoxide in ring IV, not an aziridine or azetidine as previously reported. PMID- 16357992 TI - The small peptide-catalyzed direct asymmetric aldol reaction in water. AB - Simple modular di- and tripeptides with a primary amine at the N-terminus catalyze the aqueous asymmetric aldol reaction between unmodified ketones and aldehydes to furnish the corresponding beta-hydroxy ketones with up to 86% ee in water and 99% ee in aqueous media. PMID- 16357993 TI - Asymmetric allylic oxidation reactions catalyzed by a chiral nonracemic and C2 symmetric 2,2'-bipyridyl copper(I) complex. AB - The evaluation of a chiral, nonracemic and C2-symmetric 2,2'-bipyridyl ligand in copper(I)-catalyzed asymmetric allylic oxidation reactions of a series of cyclic alkenes with tert-butyl peroxybenzoate is reported (up to 91% ee, the highest reported enantioselectivity for a bipyridyl ligand copper(I) complex to date). PMID- 16357994 TI - One-pot chemo-enzymatic synthesis of reporter-modified proteins. AB - To meet recent advancements in the covalent reporter labeling of proteins, we propose a flexible synthesis for reporter analogs. Here we demonstrate a one-pot chemo-enzymatic synthesis of reporter-labeled proteins that allows the covalent tethering of any amine-terminal fluorescent or affinity label to a carrier protein or fusion construct. This two-reaction sequence consists of activated panthothenate coupling, biosynthetic conversion to the coenzyme A (CoA) analog, and enzymatic carrier protein modification via phosphopantetheinyltransferase (PPTase). We also probe substrate specificity for CoAA, the first enzyme in the pathway. With this approach CoA analogs may be rapidly prepared, thus permitting the regiospecific attachment of reporter moieties from a variety of molecular species. PMID- 16357995 TI - Highly efficient methods for the one-pot synthesis of beta-substituted enones. AB - A mild and practically-convenient one-pot procedure for the direct beta substitution of enones has been developed using a conjugate addition-oxidation strategy with a full range of copper-based reagents and N-tert butylphenylsulfinimidoyl chloride; alkyl- and aryl-substituted enones are delivered in good to excellent yields. PMID- 16357996 TI - Stereoselective synthesis of 2,4,5-trisubstituted piperidines by carbonyl ene and Prins cyclisations. AB - Cyclisation of aldehydes 3-e catalysed by concentrated hydrochloric acid affords predominantly the all cis 2,4,5-trisubstituted piperidines 4a-ewhen the 2 substituent is small, while catalysis by MeAlCl2 in refluxing chloroform gives the trans piperidines 5a-e with diastereomeric ratios of up to 99:1. PMID- 16357997 TI - A new approach to combretastatin D2. AB - A concise and convergent route to combretastatin D2 is described together with some preliminary biological data. PMID- 16357998 TI - Organocatalytic asymmetric allylic carbon-carbon bond formation. AB - Organocatalytic allylic C-C bond-forming addition of activated alkylidenes to alkyl and aryl nitroalkenes has been achieved with high diastereo- and enantioselectivity. Chiral tertiary amine catalysts are used to give allyl intermediates which exhibit gamma-selectivity in the C-C bond forming step. The reactions proceed with up to >99:1 syn:anti ratio for both the alkyl- and aryl nitroalkenes with up 96% and 98% ee, respectively. The products of this conjugate addition are transformed into a range of intermediates, such as optically active conjugated dienes and 1-substituted tetralones, which are difficult to access via alternative methods. PMID- 16357999 TI - Mechanism of 1,4,5,8-naphthalene tetracarboxylic acid dianhydride hydrolysis and formation in aqueous solution. AB - The study of highly conjugated, carbonyl-containing molecules such as 1,4,5,8 naphthalene tetracarboxylic dianhydride, III, is of interest since reactivity differences and transmission of electronic effects through the conjugated framework can be evidenced. The kinetics of hydrolysis of III in aqueous solution were determined from 5 M acid to pH 10. In basic solution hydrolysis of III yields, sequentially, 1,4,5,8-naphthalene diacid monoanhydride, II, and 1,4,5,8 naphthalene tetracarboxylic acid, I. The second order rate constant for alkaline hydrolysis is 200 fold higher for the first ring opening. The water-catalyzed hydrolysis of III yields a pH-dependent mixture of ionic forms of I and II. The rate constant for water-catalyzed hydrolysis of III is 25 fold higher than that for II. In concentrated acid the rates for reaching equilibrium (I, II and III) increase and III is the major product. The pK(a)s of I (3.24, 5.13 and 6.25) and II (3.05, 5.90) were determined by potentiometric, fluorescence and UV spectroscopy titrations and by quantitative fit of the kinetic and equilibrium data. The apparent, pH-dependent, equilibrium constants, K(EqII), for anhydride formation between I and II were obtained from the UV spectra. The quantitative fit of kinetic and equilibrium data are consistent with the assumption that anhydride formation only proceeds with the fully protonated species for both I and II and permitted the estimation of the equilibrium constants for anhydride formation, K(EqII). The value of K(EqII) (I <==> II) between pH 1 and 6 was ca. 5. Geometry optimization calculations in the gas phase of the reactions of III in alkaline, neutral and acid conditions, at the DFT level of theory, gave electronic distributions that were qualitatively consistent with the experimental results. PMID- 16358000 TI - Activity of synthetic ion channels is influenced by cation-pi interactions with phospholipid headgroups. AB - A suite of synthetic hydraphile ion channels has been used to probe the possibility of cation-pi interactions between the channel and the phospholipid bilayer. The hydraphiles selected for this study contained either no sidearm, aliphatic sidearms or aromatic sidearms that varied in electron-richness. An ion selective electrode (ISE) method was used to evaluate the ion transport ability of these hydraphiles across synthetic bilayers. Transport was dependent on sidearm identity. Ion transport activity for the aromatic sidechained compounds was greatest when the sidearms were electron rich and vesicles were prepared from 100% DOPC (trimethylammonium cation headgroup, overall neutral). When the lipid headgroups were made more negative by changing the composition from DOPC to 70 : 30 (w/w) DOPC : DOPA, transport by the aromatic-sidechained channels was reduced. Fluorescence studies showed that when the lipid composition changed, the headgroups experienced a different polarity, suggesting reorientation. The data are in accord with a stabilizing cation-pi interaction between the aromatic sidearm of the hydraphile channel and the ammonium phospholipid headgroup. PMID- 16358001 TI - A promiscuous glutathione transferase transformed into a selective thiolester hydrolase. AB - Human glutathione transferase A1-1 (hGST A1-1) can be reengineered by rational design into a catalyst for thiolester hydrolysis with a catalytic proficiency of 1.4 x 10(7) M(-1). The thiolester hydrolase, A216H that was obtained by the introduction of a single histidine residue at position 216 catalyzed the hydrolysis of a substrate termed GSB, a thiolester of glutathione and benzoic acid. Here we investigate the substrate requirements of this designed enzyme by screening a thiolester library. We found that only two thiolesters out of 18 were substrates for A216H. The A216H-catalyzed hydrolysis of GS-2 (thiolester of glutathione and naphthalenecarboxylic acid) exhibits a k(cat) of 0.0032 min(-1) and a KM of 41 microM. The previously reported catalysis of GSB has a k(cat) of 0.00078 min(-1) and KM of 5 microM. The k(cat) for A216H-catalyzed hydrolysis of GS-2 is thus 4.1 times higher than for GSB. The catalytic proficiency (k(cat)/KM)/k(uncat) for GS-2 is 3 x 10(6) M(-1). The promiscuous feature of the wt protein towards a range of different substrates has not been conserved in A216H but we have obtained a selective enzyme with high demands on the substrate. PMID- 16358002 TI - Design and synthesis of iridium(III) azacrown complex: application as a highly sensitive metal cation phosphorescence sensor. AB - A new metal cation probe bearing a central Ir(III) element and 1-aza-15-crown-5 ether substituted pyridyl pyrazolate as the chelate was synthesized. The octahedral molecular structure of was confirmed using single crystal X-ray diffraction analyses. Subsequent photophysical study showed yellow-green emission at approximately 560 nm in both fluid solution and solid state at room temperature. Remarkable differentiation in spectral properties upon metal cation (e.g. Ca2+) complexation makes complex a highly sensitive phosphorescence probe. PMID- 16358003 TI - Rapid and efficient synthesis of poly-substituted quinolines assisted by p toluene sulphonic acid under solvent-free conditions: comparative study of microwave irradiation versus conventional heating. AB - A rapid and efficient method for the preparation of various poly-substituted quinolines has been developed through the Friedlander condensation of 2 aminoarylketone or 2-aminoarylaldehyde with carbonyl compounds in the presence of p-toluene sulphonic acid, which was achieved by both microwave irradiation and conventional heating under solvent-free conditions. PMID- 16358004 TI - One-pot Pd/C catalysed 'domino' HALEX and Sonogashira reactions: a ligand- and Cu free alternative. AB - The advantages of combining heterogeneous catalysis and aryl chloride substrates for cross-coupling are introduced. A heterogeneous Pd/C catalyst is used for activating aryl bromides and electron withdrawing aryl chlorides via a one-pot 'domino' HALEX-Sonogashira reaction. No ligand or co-catalyst is required, and the cross-coupling products are obtained in moderate to good yields. The influence of the solvent, base, iodide source and catalyst is evaluated. The catalyst is reusable for at least six consecutive reaction cycles. A variation on this reaction using catalytic amounts of KI is also proposed. PMID- 16358005 TI - Borrowing hydrogen: iridium-catalysed reactions for the formation of C-C bonds from alcohols. AB - Alcohols have been employed as substrates for C-C bond-forming reactions which involve initial activation by the temporary removal of hydrogen to form an aldehyde. The intermediate aldehyde is converted into an alkene via a Horner Wadsworth-Emmons reaction, nitroaldol and aldol reactions. The 'borrowed hydrogen' is then returned to the alkene to form a C-C bond. PMID- 16358006 TI - Molecular iodine: a highly efficient catalyst in the synthesis of quinolines via Friedlander annulation. AB - A mild and efficient route for the synthesis of quinolines and polycyclic quinolines via Friedlander annulation, utilizing molecular iodine (1 mol%) as a new catalyst, is described. PMID- 16358007 TI - Iminophosphorane-mediated efficient synthesis of new tricyclic 3,5-dihydro-1,2,3 triazolo[4,5-d]-1,2,4-triazolo[1,5-a]pyrimidin-9-ones. AB - The carbodiimides 2, obtained from aza-Wittig reactions of iminophosphorane 1 with aromatic isocyanates, reacted with hydrazine to give selectively 6-amino-7H 1,2,3-triazolo[4,5-d]pyrimidin-7-ones 5. Compounds 5 were further transformed to iminophosphoranes 6 by reaction with triphenylphosphine, hexachloroethane and triethylamine. A tandem aza-Wittig reaction of iminophosphorane 6 with isocyanate or acyl chloride generated previously unreported 3,5-dihydro-1,2,3-triazolo[4,5 d]-1,2,4-triazolo[1,5-a]pyrimidin-9-ones 10 or 12 in satisfactory yield. X-ray structure analysis of 10 g verified the proposed structure and the reaction selectivity. PMID- 16358008 TI - Nucleophilic identity substitution reactions. The reaction between hydrogen fluoride and protonated alkyl fluorides. AB - The gas phase reactions between HF and the protonated alkyl fluorides MeFH+, EtFH+, Pr(i)FH+, and Bu(t)FH+ have been studied using ab initio methods. The potential energy profiles for both nucleophilic substitution (S(N)2) and elimination (E2) pathways have been investigated. Both backside Walden inversion and frontside nucleophilic substitution reaction profiles have been generated. Backside substitution is very favourable, but shows relatively little variation with the alkyl group. Frontside substitution reaction barriers are only slightly higher than the barrier for backside substitution for HF + MeFH+, and the difference in barrier heights for frontside and backside displacement seems negligible for the larger alkyl groups. Reaction barrier trends have been analysed and compared with the results of similar studies of the H2O/ROH2+ and NH3/RNH3+ systems (R = Me, Et, Pr(i), and Bu(t)). Compared to the two other classes, protonated fluorides have extreme structures which, with the exception of the Me substrate, are weakly bound complexes between an alkyl cation and HF. The results nourish the idea that nucleophilic substitution reactions are better understood in view of competition between frontside and backside substitution than from the traditional S(N)1/S(N)2 perspective. PMID- 16358009 TI - Chemoenzymatic synthesis of GM3 and GM2 gangliosides containing a truncated ceramide functionalized for glycoconjugate synthesis and solid phase applications. AB - Analogues of GM3 and GM2 gangliosides were chemoenzymatically synthesized on a multifunctional ceramide-type tether designed to facilitate diverse strategies for glycoconjugate synthesis. The truncated ceramide aglycon maintains the stereogenic centres of natural ceramide while avoiding extensive hydrophobicity that can hamper synthesis and purification of the glycolipids. Tetanus toxoid and BSA glycoconjugates of these two gangliosides were prepared for immunization of mice, and for solid phase assays to screen for ganglioside-specific antibodies. Inhibition experiments showed that antibodies generated by tetanus toxoid conjugates of GM3 and GM2 exhibited specificity for the carbohydrate epitope and the stereogenic centres of the ceramide. PMID- 16358010 TI - Synthesis of enantiopure C3- and C4-hydroxyretinals and their enzymatic reduction by ADH8 from Xenopus laevis. AB - (R)-all-trans-3-hydroxyretinal 1, (S)-all-trans-4-hydroxyretinal and (R)-all trans-4-hydroxyretinal have been synthesized stereoselectively by Horner Wadsworth-Emmons and Stille cross-coupling as bond-forming reactions. The CBS method of ketone reduction was used in the enantioface-differentiation step to provide the precursors for the synthesis of the 4-hydroxyretinal enantiomers. The kinetic constants of Xenopus laevis ADH8 with these retinoids have been determined. PMID- 16358011 TI - Computer simulation of polypeptide adsorption on model biomaterials. AB - When biomaterials are inserted in a biological environment, for instance in a body implant, proteins do quickly adsorb on the exposed surface. Such process is of fundamental importance, since it directs the subsequent cell adhesion. Here we review recent advances in this field obtained with molecular simulations. While coarse-grained models can provide important general results, as it has long been recognized in polymer science, the hierarchical structure of a very complex copolymer such as a protein, together with the nature of the biomaterial surface suggest that atomistic models are better suited to investigate these phenomena. Thus, after briefly mentioning some common features of coarse-grained and atomistic force fields, we first discuss early theoretical and coarse-grained simulation results about protein adsorption, and then we highlight the main results recently obtained by us with atomistic models. In particular, we discuss some conformational and energetic aspects of the adsorption of protein fragments with different secondary structure on surfaces of different wettability, including hydrophobic graphite and hydrophilic poly(vinylalcohol). We also consider other features, such as the simulation of the materials wettability, the hydration of the adsorbed fragments, their kinetics of spreading, and the sequential adsorption of two protein fragments on top of each other, highlighting the results of general interest. PMID- 16358012 TI - Molecular structure calculations without clamping the nuclei. AB - A number of recent papers have considered ways in which molecular structure may be calculated when both the electrons and the nuclei are treated from the outset as quantum particles. This is in contrast to the conventional approach in which the nuclei initially have their positions fixed and so merely provide a potential for electronic motion. The usual approach is generally assumed to be justified by the 1927 work of Born and Oppenheimer. In this paper we discuss what precisely might be anticipated in the way of molecular structure from a mathematical consideration of the spectral properties of the full Coulomb Hamiltonian, to what extent the very idea of molecular structure might be dependent upon treating the nuclei simply as providing a potential and the extent to which the work of Born and Oppenheimer can be used to support this position. PMID- 16358013 TI - The effect of torsion angle on the rate of intramolecular triplet energy transfer. PMID- 16358014 TI - Rotationally resolved electronic spectroscopy of tryptophol in the gas phase. AB - High resolution S1-S0 fluorescence excitation spectra of tryptophol have been observed in the collision-free environment of a supersonic beam. Each origin band has been assigned to a unique conformer based on its observed rotational constants. Unlike its close relative tryptamine, which exhibits seven distinguishable conformers under similar conditions, tryptophol exhibits only four (GPy-in, GPh-in, and two anti structures). Possible reasons for this difference in behavior are discussed. PMID- 16358015 TI - The high resolution inelastic neutron scattering spectrum of ammonium fluoride. AB - The measured high resolution (deltaE/E approximately 2-3%) incoherent inelastic neutron scattering spectrum of ammonium fluoride is presented and discussed with reference to the available optical spectra. In addition, a full set of dispersion curves have been obtained from a new ab initio lattice dynamics calculation and these have been used to produce a rigorous interpretation of the spectrum. The librational modes of the ammonium ion occur at 560 cm(-1) and the anharmonicity in these modes is estimated to be 4%, about half that observed in the other ammonium halides. The reduction in anharmonicity is attributed to stronger hydrogen bonding and a deeper potential well. The calculations agree well with the observed spectrum apart from the librational modes which are shifted upwards by around 40 cm(-1) from the measured values. Dispersion and LO-TO splitting are important in this system with modes changing frequency by up to 135 cm(-1). The nature of the calculated LO-TO splitting in the optic mode region is indicative of a pseudo-cubic system confirming that the site symmetry of the ammonium ion is very close to T(d). Because of LO-TO splitting the ammonium ion asymmetric stretch, nu3, has components calculated to be at higher frequencies than those of the symmetric stretch, nu1, which contradicts the assignment scheme produced from optical data. PMID- 16358016 TI - Calculation of adiabatic polarization of atomic photofragments under the influence of long range quadrupole-quadrupole interactions. AB - General equations for diagonalization of the quadrupole-quadrupole interaction matrix for diatomic molecules are presented. Eigenvalue and eigenvector solutions are tabulated for atoms with levels J < or = 2. The use of the eigenvector solutions for determination of adiabatic molecule-frame photofragment polarization is illustrated, and polarization parameters a and a are tabulated. Even if knowledge of the photofragment scattering matrix is limited, we illustrate how coherent polarization parameters a0(1) and a0(2) can be calculated from the tabulated adiabatic expansion coefficients, for example, by making use of available experimental data. PMID- 16358017 TI - A temperature-dependent relative-rate study of the OH initiated oxidation of n butane: the kinetics of the reactions of the 1- and 2-butoxy radicals. AB - The kinetics of the reactions of 1-and 2-butoxy radicals have been studied using a slow-flow photochemical reactor with GC-FID detection of reactants and products. Branching ratios between decomposition, CH3CH(O*)CH2CH3 --> CH3CHO + C2H5, reaction (7), and reaction with oxygen, CH3CH(O*)CH2CH3+ O2 --> CH3C(O)C2H5+ HO2, reaction (6), for the 2-butoxy radical and between isomerization, CH3CH2CH2CH2O* --> CH2CH2CH2CH2OH, reaction (9), and reaction with oxygen, CH3CH2CH2CH2O* + O2 --> C3H7CHO + HO2, reaction (8), for the 1-butoxy radical were measured as a function of oxygen concentration at atmospheric pressure over the temperature range 250-318 K. Evidence for the formation of a small fraction of chemically activated alkoxy radicals generated from the photolysis of alkyl nitrite precursors and from the exothermic reaction of 2 butyl peroxy radicals with NO was observed. The temperature dependence of the rate constant ratios for a thermalized system is given by k7/k6= 5.4 x 10(26) exp[(-47.4 +/- 2.8 kJ mol(-1))/RT] molecule cm(-3) and k9/k8= 1.98 x 10(23) exp[( 22.6 +/- 3.9 kJ mol(-1))/RT] molecule cm(-3). The results agree well with the available experimental literature data at ambient temperature but the temperature dependence of the rate constant ratios is weaker than in current recommendations. PMID- 16358018 TI - Mechanism of HCS + O2 reaction: hydrogen- or oxygen-transfer? AB - In spite of the potential importance of the HCS radical in both combustion and interstellar processes, its chemical reactivity has not been tackled previously. In the present paper, the oxidation reaction of the HCS radical is theoretically investigated for the first time at the CCSD(T)/6-311++G(3df,2p)//BH&HLYP/6 311++G(d,p)+ZPVE and Gaussian-3//B3LYP/6-31G(d) levels. It is shown that the most feasible pathway is the O2 addition to the HCS radical forming the intermediate SC(H)OO which can undergo a subsequent O-extrusion leading to SC(H)O + 3O. This features an indirect O-transfer mechanism with the overall barrier of 4.4 and 3.5 kcal mol(-1), respectively, at the two levels. However, formation of the H transfer product CS + HO2 is kinetically much less feasible, i.e., the direct mechanism has barriers of 14.3 and 8.7 kcal mol(-1), whereas the indirect mechanism has barriers of 12.6 and 10.7 kcal mol(-1), respectively. This result is in sharp contrast to the analogous HCO + O2 reaction, where the direct (with a barrier of 2.98 kcal mol(-1)) and indirect (2.26 kcal mol(-1)) H-transfer processes are highly competitive over the indirect O-transfer process (the least endothermicity is 19.9 kcal mol(-1)). The possible explanations and implications of the present results are provided. PMID- 16358019 TI - Inclined N2 desorption in N2O reduction by D2 and CO on Pd(110). AB - Inclined N2 desorption was examined in the course of a catalyzed N2O + D2 (or CO) reaction on Pd(110) by angle-resolved mass spectroscopy combined with cross correlation time-of-flight techniques. N2 desorption collimated at around 45 degrees off the normal toward the [001] direction in the temperature range of 400 800 K. Its collimation angle and kinetic energy were insensitive to both the surface temperature and surface conditions throughout the kinetic transition. It is proposed that this peculiar N2 desorption is induced by the decomposition of N2O oriented along the [001] direction. PMID- 16358020 TI - Experimental study and Monte-Carlo simulation of the nucleation and growth processes during the dehydration of Li2SO4.H2O single crystals. AB - A kinetic model for the dehydration of lithium sulfate monohydrate is proposed in order to account for experimental data obtained on single crystals by thermogravimetry at 80 degrees C under fixed water vapour pressure, and by optical microscopy. This model is based on the assumptions of Mampel's model, the nucleation takes place randomly at the surface of the solid and is followed by isotropic growth toward the centre of the crystal. Calculated rates dalpha/dt are obtained by means of Monte-Carlo simulations and compared to the experimental ones, which leads to the determination of two kinetic constants: the areic frequency of nucleation (in number of nuclei m(-2) s(-1)) and the areic reactivity of growth (in mol m(-2) s(-1)). PMID- 16358021 TI - Artificial multiple criticality and phase equilibria: an investigation of the PC SAFT approach. AB - The perturbed-chain statistical associating fluid theory (PC-SAFT) is studied for a wide range of temperature, T, pressure, p, and (effective) chain length, m, to establish the generic phase diagram of polymers according to this theory. In addition to the expected gas-liquid coexistence, two additional phase separations are found, termed "gas-gas" equilibrium (at very low densities) and "liquid liquid" equilibrium (at densities where the system is expected to be solid already). These phase separations imply that in one-component polymer systems three critical points occur, as well as equilibria of three fluid phases at triple points. However, Monte Carlo simulations of the corresponding system yield no trace of the gas-gas and liquid-liquid equilibria, and we conclude that the latter are just artefacts of the PC-SAFT approach. Using PC-SAFT to correlate data for polybutadiene melts, we suggest that discrepancies in modelling the polymer density at ambient temperature and high pressure can be related to the presumably artificial liquid-liquid phase separation at lower temperatures. Thus, particular care is needed in engineering applications of the PC-SAFT theory that aims at predicting properties of macromolecular materials. PMID- 16358022 TI - The low-temperature proton-ordered phases of ice predicted by ab initio methods. AB - The low-temperature proton-ordered counterparts for ice-Ih, ice-III, ice-VI and ice-VII are investigated by first principle methods in conjunction with a graph enumeration technique. Two experimentally well calibrated disorder/order transitions, ice-Ih/ice-XI and ice-VII/ice-VIII, are used to validate the methodology we used herein and in both cases our approach is able to reproduce major experimental features. For ice-III and ice-VI, direct structural determination on the fully proton-ordered counterparts by neutron diffraction is not available and in the literature different ordering schemes have been proposed. Our calculation results serve as an independent reference, and we shall discuss our findings and their relevance to previous experimental works. PMID- 16358023 TI - The electron transfer rate of large TPA based compounds: a joint theoretical and electrochemical approach. AB - A series of triphenylamine (TPA) based compounds is investigated by means of density functional theory and cyclic voltammetry. Using the Nicholson's formalism, the measured deltaE(p) are correlated with B3LYP/6-31G* calculated reorganisation energies (lambda), elucidating the trend followed by the electron transfer rate of these compounds. Besides the direct dependency upon the dimension of the cationic fragment contributing to the hole stabilisation, the lambdas are tuned by the symmetry local to the TPA units, as evidenced by the structural relaxation of the cations. MDTAB shows the interesting combination of low ionisation potential (IP) and low lambda. This can make this compound interesting for practical applications in organic light emitting diode (OLEDs) devices, due to the direct correlation of the IP and lambda with the hole transfer efficiency to the anode, along with the hole mobility. PMID- 16358024 TI - The low energy tautomers and conformers of the dipeptides HisGly and GlyHis and of their sodium ion complexes in the gas phase. AB - The low-lying conformers of the dipeptides HisGly and GlyHis, and of their sodium cation complexes, have been studied with a combination of Monte Carlo search with the Amber force field and local geometry optimization at the ab initio HF/6 31G(d) level, completed with MP2(full)/6-311+G(2d,2p) energetics at the HF/6 31G(d) geometries. For each dipeptide, both the N(delta)-H and N(epsilon)-H tautomers of the imidazole side chain of His were considered. For each of the four isomeric dipeptides, 20-30 conformers were fully characterized at the ab initio level. All low energy structures are found to involve H-bonding at the N(delta) position of imidazole, either as a N-H donor or a N acceptor, depending upon the tautomer. In three out of the four species, the most stable conformer involves a C-terminus carboxylic acid in its less favorable trans conformation, in order to maximize intramolecular H bonding. It turns out that the lowest energy tautomer of HisGly is N(epsilon)-H, while that of GlyHis is N(delta)-H. This result argues in favor of the diversity of His tautomeric states in peptides and proteins. The sodium cation complexes of both GlyHis and HisGly have been studied as well, again considering both tautomers in each case. In three out of the four species, the most stable structure involves chelation of sodium by the two carbonyl oxygens and the imidazole ring. On the contrary, the sodium complex of the N(delta)-H tautomer of HisGly favors chelation to the peptidic carbonyl oxygen, the imidazole ring and the amino terminus. In the N(epsilon)-H tautomers of both peptides, the most favorable binding site of imidazole is the N(delta) nitrogen, while in the N(delta)-H tautomers, it is the pi cloud which provides side chain interaction. As a result, both GlyHisNa+ and HisGlyNa+ favor the N(epsilon)-H tautomer of His, in contrast to what was found for the free peptides. PMID- 16358025 TI - Structure and reactivity of C54q+ (q = 0, 1, 2 and 4) fullerenes. AB - Using density functional theory we have studied the structural properties of eleven C54 isomers that appear in the C60 fragmentation. We have evaluated the relative stability of the different isomers with respect the most stable one, which corresponds to the structure with the minimum number (four) of adjacent pentagons. On average, the length of a bond shared by pentagons and/or hexagons increases in the order hexagon-hexagon, hexagon-pentagon and pentagon-pentagon. However, we have found that the central bond in the confluence of four hexagons, i.e. a pyrene substructure, is anomalously large, becoming in some cases the largest one. We have also evaluated the nucleus-independent chemical shifts (NICS) at the center of every individual ring in the most stable isomers. For the chlorine derivatives, our calculations show that the favorite position for chlorine addition are the bonds shared by pentagons. PMID- 16358026 TI - Redox and redox-coupled processes of heme proteins and enzymes at electrochemical interfaces. AB - Modern bioelectrochemical methods rely upon the immobilisation of redox proteins and enzymes on electrodes coated with biocompatible materials to prevent denaturation. However, even when protein denaturation is effectively avoided, heterogeneous protein electron transfer is often coupled to non-Faradaic processes like reorientation, conformational transitions or acid-base equilibria. Disentangling these processes requires methods capable of probing simultaneously the structure and reaction dynamics of the adsorbed species. Here we provide an overview of the recent developments in Raman and infrared surface-enhanced spectroelectrochemical techniques applied to the study of soluble and membrane bound redox heme proteins and enzymes. Possible biological implications of the findings are critically discussed. PMID- 16358027 TI - Computational studies of the cationic aluminium(chloro) hydroxides by quantum chemical ab initio methods. AB - Cationic aluminium(chloro) hydroxide complexes with two to four aluminium atoms were studied using quantum chemical methods. Complexes were studied in both gas and liquid phase. The liquid environment was modeled by using a conductor-like screening model (COSMO). COSMO calculations were carried out as a single point calculation at the optimized gas phase structures. Water (epsilon = 78.54) was used as the solvent. The minimum energy structures obtained from the gas phase studies were mostly compact cyclic structures. Aluminium preferred to be five coordinated in oxygen rich clusters. Core oxygen preferred three-fold coordination but in the largest clusters the four-coordinated oxygen was observed. Water reacted dissociatively with hydrogen poor clusters. The COSMO calculations showed that the optimal structures of cationic aluminium(chloro) hydroxides tend to be more open in the liquid than in the gas phase. PMID- 16358028 TI - Structures and infrared spectra of fluoride-hydrogen sulfide clusters from ab initio calculations: F(-)-(H2S)n, n = 1-5. AB - Clusters formed between a fluoride anion and several hydrogen sulfide molecules have been investigated via ab initio calculations at the MP2 level of theory, using Dunning's augmented correlation consistent basis sets. Optimised geometries, vibrational frequencies, and enthalpy changes for the ligand association reactions are presented for clusters with up to five H2S ligands interacting with a F- anion. The minimum energy structure for the 1:1 F(-)-H2S complex features proton transfer from the H2S to the F- anion, forming a planar C(s) symmetry FH...SH- structure. For the F(-)-(H2S)2 cluster, the FH...SH- core remains and is solvated by a perturbed H2S ligand. For the larger F(-)-(H2S)(3-5) clusters, in addition to the FH...SH(-)-(H2S)n cluster forms, other minima featuring a 'solvated F-' anion are predicted. Calculated infrared spectra for the minima of each cluster size are presented to aid in assigning spectra from future experimental studies. PMID- 16358029 TI - Properties of interatomic surfaces: relation to bond energies. AB - A number of properties of interatomic surfaces, as defined by the quantum theory of atoms in molecules (QTAIM), are calculated for approximately 50 molecules. These integrated surface properties are then tested for their ability to correlate and predict bond energies from MP2 atomisation energies. Three surface properties, each with units of energy, are found to show strong correlations with bond energy: single parameter models work well for non-polar bonds, but fail for polar and ionic bonds, where multi-variate methods are required. The local curvature of the interatomic surface is found to be useful this respect, reflecting charge transfer effects. PMID- 16358030 TI - Structure and stability of oxygen adsorption on Si(n) (n = 5-10) clusters. AB - The structures, binding energies, and electronic properties of one oxygen atom (O) and two oxygen atoms (2O) adsorption on silicon clusters Si(n) with n ranging from 5 to 10 are studied systematically by ab initio calculations. Twelve stable structures are obtained, two of which are in agreement with those reported in previous literature and the others are new structures that have not been proposed before. Further investigations on the fragmentations of Si(n)O and Si(n)O2 (n = 5 10) clusters indicate that the pathways Si(n)O --> Si(n-1) + SiO and Si(n)O2 --> Si(n-2) + Si2O2 are most favorable from thermodynamic viewpoint. Among the studied silicon oxide clusters, Si8O, Si9O, Si5O2 and Si8O2 correspond to large adsorption energies of silicon clusters with respect to O or 2O, while Si8O, with the smallest dissociation energy, has a tendency to separate into Si7 + SiO. Using the recently developed quasi-atomic minimal-basis-orbital method, we have also calculated the unsaturated valences of the neutral Si(n) clusters. Our calculation results show that the Si atoms which have the largest unsaturated valences are more attractive to O atom. Placing O atom right around the Si atoms with the largest unsaturated valences usually leads to stable structures of the silicon oxide clusters. PMID- 16358031 TI - The (010) surface of alpha-MoO3, a DFT + U study. AB - We apply periodic density functional theory to alpha-MoO3 and its (010) surface. The formation energy and structure of defects in the form of surface oxygen vacancies are found to depend critically on the treatment of electron localisation which is achieved in the periodic model using the DFT + U method. Calculated vibrational states for the defect free surface are found to agree well with surface science experimental data and we show that the molybdenyl stretching mode is shifted to a lower frequency in the neighbourhood of a terminal oxygen vacancy. Adsorption of molecular oxygen at the defect site can result in O2, O2( ) or O2(2-) surface species depending on the geometry of adsorption. PMID- 16358032 TI - Theory of the photodissociation of ozone in the Hartley continuum; effect of vibrational excitation and O(1D) atom velocity distribution. AB - The effect of vibrational excitation on the photodissociation cross section of ozone in the Hartley continuum is examined. The calculations make use of newly computed potential energy and transition dipole moment surfaces. The initial vibrational states of the ozone are computed using grid based techniques and the first few ab initio computed vibrational energy level spacings agree to within 10 cm(-1) with experimental values. The computed total absorption cross sections arising from different initial vibrational states of ozone are discussed in the light of the nature of the transition dipole moment surface. The computed cross section for excitation from the ground vibrational-rotational state is in good agreement with the experimentally measured cross section. Excitation of the asymmetric stretching vibration of ozone has a marked effect on both the form and magnitude of the photodissociation cross section. The velocity distributions of highly reactive O(1D) atoms arising from the photodissociation process in different wavelength ranges is also presented. The results show that the O(1D) atoms travel with a most probable translational velocity of 2.030 km s(-1) corresponding to a translational energy of 0.342 eV or 33.0 kJ mol(-1). PMID- 16358033 TI - Infrared spectra and structures of the Th(OH)2 and Th(OH)4 molecules. AB - Thorium atoms react with H2O2, H2 + O2 mixtures, and H2O in excess argon to form the Th(OH)2 and Th(OH)4 molecules as minor and major products, respectively. The vibrational frequencies observed in the matrix infrared spectra are in excellent agreement with MP2 computed values, which confirms the identification of these highly ionic thorium hydroxide molecules. Our MP2 calculations converge to slightly bent and tetrahedral structures, respectively. This investigation reports the first evidence for pure actinide dihydroxide and tetrahydroxide molecules. PMID- 16358034 TI - Behaviour of Pluronic P84 block copolymer micelles above the gelification temperature as probed by positron annihilation lifetime spectroscopy. AB - The new method based on positron annihilation lifetime spectroscopy (PALS) to determine both the mean core radius, R(core), and aggregation number, N(ag), of micelles is applied to the study of aqueous solutions of the triblock Pluronic P84 copolymer as a function of temperature (T), beyond the gelification point (334 K). Two long-lived components appear in the PALS spectra, ascribed to triplet positronium in the water bulk (o-Ps(aq)) and in the organic core of the micelles (o-Ps(org)). Of the various fitting parameters, only the lifetime of the latter species, tau4, and the micellar parameters, R(core) and N(ag), disclose the occurrence of gelification by first increasing up to 334 K, then decreasing. By contrast to what is known in case of phase transition, none of the parameters shows any abrupt change at 334 K, whereas the macroscopic viscosity of the solutions suffers a drastic increase. This is attributed to the fact that positronium is sensitive to the microviscosity of the solutions. At the transition point, the properties of the polyoxipropylene aggregates forming the organic core of the P84 micelles are not greatly affected. Furthermore, the fact that the experimental N(ag) values coincide with those calculated for spheres, from the R(core) values, indicates that the shape of the P84 cores does not change significantly after gelification. The onset of gelification results from a decrease in the hydrogen bonding interactions in the solution with an ensuing relative increase in the interactions between the polyoxipropylene (PPO) groups, initially forming the corona of the P84 micelles, in an intermicellar mode. This increased solicitation of the PPO groups outside their initial location would result in depletion in the number of surfactant molecules forming the micelles, viz. a decrease in both R(core) and N(ag) above 334 K. From the data, additional information can be gained regarding the local viscosity and surface tension in the micellar cores. PMID- 16358035 TI - Regular Liesegang patterns and precipitation waves in an open system. AB - We investigate the regular and moving Liesegang pattern formation phenomena in an open system. First, simulations have been performed at fixed coupling between the reactive medium and the reservoir, later this control parameter was varied during the simulations resulting in various phenomena. We predicted and monitored for the first time various--dynamically changing--precipitation structures and a spatial hysteresis phenomenon, which is beyond the scope of the Turing instability. The dynamics of the reaction is well detectable using specific quantities: the total amount of precipitate and its center of gravity. PMID- 16358036 TI - Electron transport in silver-semiconductor nanocomposite films exhibiting multicolor photochromism. AB - In the multicolor photochromism of TiO2 nanoporous films loaded with photocatalytically deposited Ag nanoparticles, visible light-induced electron transfer from Ag to oxygen molecules plays an essential role. Here we examined the effect of TiO2 on the electron transfer. We found that not only photocatalytically deposited Ag, but also electrodeposited Ag and commercially available Ag nanoparticles in a nanoporous TiO2 film exhibit the multicolor photochromism. The electrodeposited Ag exhibits the multicolor photochromism also in a nanoporous ZnO film, but not in nanoporous indium-tin oxide (ITO) and SiO2 matrices. Photoelectrochemical measurements for the Ag-TiO2 nanocomposite elucidated that some of the photo-excited electrons on Ag are transferred to oxygen molecules via TiO2 and non-excited Ag. Thus, an n-type semiconductor plays an important role in the charge separation between the excited electrons and Ag+. Non-excited Ag on TiO2 also plays an important role in the charge separation and/or catalysis of oxygen reduction. Replacement of the non-excited Ag with Pt accelerated the electron transport from the photo-excited Ag to oxygen molecules and the photochromic behavior. PMID- 16358037 TI - Water molecule clusters measured at water/air interfaces using atomic force microscopy. AB - During the tip approach to hydrophobic surfaces like the water/air interface, the measured interaction force reveals a strong attraction with a range of approximately 250 nm at some points along the interface. The range of this force is approximately 100 times larger than the measured for gold (approximately 3 nm) and 10 times larger than the one for hydrophobic silicon surfaces (approximately 25 nm). At other points the interface exerts a medium range repulsive force growing stepwise as the tip approaches the interface plane, consequently the hydrophobic force is a strong function of position. To explain these results we propose a model where the force on the tip is associated with the exchange of a small volume of the interface with a dielectric permittivity epsilon(int) by the tip with a dielectric permittivity epsilon(tip). By assuming a oscillatory spatial dependence for the dielectric permittivity it is possible to fit the measured force profiles. This dielectric spatial variation was associated with the orientation of the water molecules arrangement in the interfacial region. Small nanosized hydrogen-bond connected cages of water molecules present in bulk water at the interface are oriented by the interfacial electric field generated by the water molecules broken bonds, one broken hydrogen bond out of every four. This interfacial field orients small clusters formed by approximately 100 water molecules into larger clusters (approximately 100 nm). In the limit of small (less than 5 nm thick) water molecule cages we have modeled the static dielectric permittivity (epsilon) as the average response of those cages. In these regions the dielectric permittivity for water/air interfaces decreases monotonically from the bulk value epsilon approximately 80 to approximately 2 at the interface. For regions filled with medium size cages, the tip senses the structure of each cage and the static dielectric permittivity is matched to the geometrical features of these cages sized approximately 25 to 40 nm. Interfacial electric energy density values were calculated using the electric field intensity and the dielectric permittivity obtained by the fitting of the experimental points. The integration of the electric energy density along the interfacial region gives a value of 0.072 J m(-2) for interfacial energy density for points where the hydrophobic force has a range of approximately 250 nm. Regions formed by various clusters result in lower values of the interfacial energy density. PMID- 16358038 TI - The interaction of CO2 with sodium-promoted W(011). AB - The activation of CO2 by interaction with Na atoms on tungsten was studied in a joint experimental/theoretical effort combining MIES, UPS (HeII) and first principles calculations. Experimentally, both the adsorption of Na on tungsten, followed by CO2 exposure to the Na-modified surface at 80 K, and the adsorption of CO2 on tungsten, followed by Na exposure to the CO2 covered substrate, were studied. Below about 120 K CO2 physisorbs on pure W(011), and the distance between the three main spectral features is as for gas phase CO2 (E(B) = 8.4, 12.1, 14.1 eV). When offered to a Na monolayer (ML) deposited onto W, CO2 is converted into a chemisorbed species. The spectral pattern is different from physisorbed CO2, and the three spectral features are shifted towards lower binding energies (E(B) = 6.3, 10.7, 13.9 eV). The chemisorption continues until all available Na species are converted into Na+ species. Additional CO2 offered to the system becomes physisorbed on top of the chemisorbed species. When a CO2 monolayer, physisorbed on tungsten at 80 K, is exposed to Na, the interaction leads initially to a decrease of the surface work function and to a rigid, global shift of all CO2 induced features towards larger binding energies by about 2 eV. Only beyond a minimum Na coverage of about 0.5 ML, chemisorbed species can be detected. We conclude that, initially, transfer of the Na(3s) electron to the tungsten substrate takes place. Above 0.5 ML Na coverage, back donation of charge to CO2 takes place whereby the physisorbed carbon dioxide species become converted into chemisorbed ones. The experimental results are interpreted with the help of first principle calculations carried out on suitable slab models. The structures and surface binding mode of the chemisorbed CO2 species are described. The calculated density of states for the most stable situations is in qualitative agreement with experimental data. PMID- 16358039 TI - Self-organization of a sulfonamido-porphyrin in Langmuir monolayers and Langmuir Blodgett films. AB - Langmuir monolayers (LM) and Langmuir-Blodgett (LB) films of pure lipophilic meso tetra(4-dodecylaminosulfophenyl)porphyrin (PC12) and mixed with the anionic surfactant sodium hexadecylsulfate (SHS) were studied. The molecular packing and structure of PC12 and PC12-4SHS with variable surface pressure were investigated by surface pressure-area measurements, steady-state absorption, fluorescence emission and anisotropy, as well as by fluorescence lifetime imaging microscopy (FLIM). At low surface pressure, the porphyrin molecules are organized with the rings tilted on the water surface whereas at high surface pressure the porphyrin rings achieve a more perpendicular arrangement. Using the FLIM images a gradual change of aggregates into large "islands" is observed. Different patterns are observed in the pure PC12 multilayer films (n = 3 and 5) with ordered patches superimposed which are not observed in the PC12-4SHS multilayer LB films. PMID- 16358040 TI - An experimental study of melting of CCl4 in carbon nanotubes. AB - We report dielectric relaxation spectroscopy measurements of the melting point of carbon tetrachloride confined within open-tip multi-walled carbon nanotubes with two different pore diameters, 4.0 and 2.8 nm. In both cases, a single transition temperature well above the bulk melting point was obtained for confined CCl4. These results contrast with what was obtained in our previous measurements using carbon nanotubes with a pore diameter of 5.0 nm, where multiple transition temperatures both above and below the bulk melting point of CCl4 were observed. Our experimental measurements are consistent with our recent molecular simulation results (F. R. Hung, B. Coasne, E. E. Santiso, K. E. Gubbins, F. R. Siperstein and M. Sliwinska-Bartkowiak, J. Chem. Phys., 2005, 122, 144706). Although the simulations overestimate the temperatures in which melting upon confinement occurs, both simulations and experiments suggest that all regions of adsorbate freeze at the same temperature, and that freezing occurs at higher temperatures upon reduction of the pore diameter. PMID- 16358041 TI - Hole-trapping effect on thermoelectric power of mixed ionic electronic conductor BaTiO3. AB - The effect of hole-trapping on the thermopower of a mixed ionic electronic conductor, e.g., BaTiO3, is analyzed in terms of irreversible thermodynamics by taking trapped holes as a fourth kind of electronic charge carrier in addition to free electrons, free holes and mobile oxide ions. It is found that the effect manifests itself in two ways: thermostatically in the ionic thermopower via the thermodynamic factor and dynamically in the electronic thermopower via the electrical conductivity contribution of the trapped holes. The thermopowers of both 99.995% pure, undoped and 1.8 m/o Al-doped BaTiO3, that were measured against oxygen activity in the range of -18 < log aO2 < or = 0 at elevated temperatures of 800 degrees to 1100 degrees C [H.-I. Yoo and C. R. Song, J. Electroceram., 2001, 6, 61, ref. 6], are reanalyzed by taking into account the hole-trapping for the doped case. It is found that while the reduced heats-of transport of free electrons and holes are, respectively, close to their thermal energy k(B)T (k(B) being the Boltzmann constant), that of trapped holes is close to their migration energy that is essentially the same as the trapping energy onto the acceptors doped, 1.04 eV. PMID- 16358042 TI - Fluorescence based strategies for genetic analysis. AB - Synthetic chemistry has been central to the design of modern methods of genetic analysis. In this article, we discuss the underlying chemistry and biophysical principles that have been used in the development of robust methods for the analysis of DNA in the diagnostic laboratory. PMID- 16358043 TI - Non-ionic polythiophenes: a non-aggregating folded structure in water. AB - The non-aggregating nature of a water-soluble pi-conjugated polythiophene has been characterised by concentration independent thermal denaturing. PMID- 16358044 TI - A chiral, heterometallic metal-organic framework derived from a tris(chelate) coordination complex. AB - A novel tris(chelate) metalloligand has been used to synthesize a chiral, heterometallic metal-organic framework that is robust to solvent removal and shows selective uptake of nitroaromatic compounds. PMID- 16358045 TI - Anion-directed self-organization of thermotropic liquid crystalline materials containing a guanidinium moiety. AB - New wedge-shaped thermotropic liquid crystalline materials containing a guanidinium moiety at the apex organize into various supramolecular structures such as hexagonal columnar, rectangular columnar and Pm3n cubic mesophases depending on anions illustrating guest-directed self-organization in mesophases. PMID- 16358046 TI - An octanuclear [Co(II)2-Co(III)2]2 interlocked grid--example of an inorganic [2]catenane. AB - An oxalic dihydrazide based flexible polydentate amidrazone ligand undergoes spontaneous [2 x 2] grid assembly, followed by partial disassembly, and then reassembly in the presence of Co(NO3)2.6H2O and NH4(NCS) to form a unique octanuclear [Co(II)2-Co(III)2]2 interlocked, single braided 4 + 4 [2]catenane. PMID- 16358047 TI - Microporous chiral metal coordination polymers: hydrothermal synthesis, channel engineering and stability of lanthanide tartrates. AB - L-Tartrate ions can endure hydrothermal conditions up to 160 degrees C to form the robust, enantiopure open-framework coordination polymers [Ln2(L TAR)3(H2O)2]3H2O, 1; the addition of succinate results in formation of the related [Ln2(L-TAR)2(SUC)(H2O)2]5.5H2O, with larger channels than , whereas racemic D/L-tartrate gives the more condensed [Ln2(D/L-TAR)3(H2O)2], . TAR = [C4H4O6]2-. PMID- 16358048 TI - CdSe/ZnS-labeled carboxymethyl chitosan as a bioprobe for live cell imaging. AB - A simple and convenient method for the construction of CdSe/ZnS-labeled polysaccharides as bioprobes were developed, which are highly biocompatible and photostable, and have been proven to be suitable for live cell imaging. PMID- 16358049 TI - Direction of unusual mixed-ligand metal-organic frameworks: a new type of 3-D polythreading involving 1-D and 2-D structural motifs and a 2-fold interpenetrating porous network. AB - The reaction of Cu(II) or Cd(II) acetate with mixed ligands terephthalic (tp) and 3,5-bis(4-pyridyl)-4-amino-1,2,4-triazole (bpt) under the same conditions affords two unusual metal-organic frameworks, in which represents a new type of polythreaded supramolecular architecture consisting of distinct 1-D and 2-D coordination polymers within one crystal, however, has an interpenetrating porous network with two similar laterally interlocking 2-D (4,4) layers. PMID- 16358050 TI - A new organometallic heteropolytungstate related to [Sb2W22O74(OH)2](12-): synthesis and structural characterisation of the bis-{Ru(p-cymene)}(2+) containing anion [Sb2W20O70{Ru(p-cymene)}2](10-). AB - The easy synthesis of [Sb2W20O70{Ru(p-cymene)}2](10-) by the reaction between the organometallic precursor [Ru(p-cymene)Cl2]2 and [Sb2W20O70]14-, formed in situ, confirms the importance of the organometallic route to well defined ruthenium(+II)-substituted heteropolytungstates. PMID- 16358051 TI - Structural relationship between a host included chain of spirocyclic water hexamers and bulk water--the role of water clusters in self assembly and crystallization processes. AB - Infinite chains of spirocyclic water hexamers are included in the crystal lattice of a tin complex with a curved, hydrophobic surface and only weak intermolecular bonding interactions between the host molecules, so that the enclosed water clusters might be reminiscent of the solvation sphere in solution. PMID- 16358052 TI - Metal directed assembly of ditopic containers and their complexes with alkylammonium salts. AB - A new self-folding cavitand has been assembled through metal coordination to give a thermodynamically stable ditopic receptor of nanosize dimensions which has been used in the reversible binding of di-alkylammonium and n-alkylammonium salts. PMID- 16358053 TI - Formation of layered titania and zirconia catalysed by surface-bound silicatein. AB - Silicatein immobilised on self-assembled polymer layers using a histidine-tag chelating anchor group retains its hydrolytical activity for the formation of biosilica, and catalyses the formation of layered arrangements of biotitania and biozirconia. PMID- 16358054 TI - Archaeosomes based on novel synthetic tetraether-type lipids for the development of oral delivery systems. AB - The in vitro stability of archaeosomes made from novel synthetic membrane spanning tetraether lipids was evaluated in conditions mimicking those of oral route application in terms of bile salts, serum and low pH.. PMID- 16358055 TI - On the influence of porphyrin pi-pi stacking on supramolecular chirality created in the porphyrin-based twisted tape structure. AB - Achiral porphyrin derivatives 1-M form twisted supramolecular assemblies and the pitch of the twisted tape structure can be altered by changing the central metal. PMID- 16358056 TI - Towards large amounts of Janus nanoparticles through a protection-deprotection route. AB - Janus silica nanoparticles, regioselectively functionalized by two different chemical groups, were synthesized through a multistep procedure based on the use of a polystyrene nodule as a protecting mask. PMID- 16358057 TI - A one pot, metathesis-hydrogenation sequence for the selective formation of carbon-carbon bonds. AB - A combination of homogeneous hydrogenation and metathesis reactions allows highly efficient, stepwise chemo- and stereoselective formation of three separate 2,7 diaminosuberic acid derivatives in a single pot without isolation of intermediates. PMID- 16358058 TI - Cross-metathesis of unsaturated natural oils with 2-butene. High conversion and productive catalyst turnovers. AB - The cross-metathesis of synthetic and natural triglycerides containing unsaturated fatty acids with 2-butene can be achieved with high conversion and excellent productive turnovers. These reactions are catalysed by second generation ruthenium-based olefin metathesis catalysts and can be conducted at -5 degrees C in liquid 2-butene. PMID- 16358059 TI - Preparation, characterization and application of magnetic silica nanoparticle functionalized multi-walled carbon nanotubes. AB - Magnetic silica nanoparticle functionalized multi-walled carbon nanotubes (MS MWNTs) were prepared, characterized and used for the convenient, rapid and efficient separation of trace aromatic compounds. PMID- 16358060 TI - Diastereoselective allylation and crotylation of N-unsubstituted imines derived from ketones. AB - A wide variety of tertiary carbinamines are synthesized in high yields via diastereoselective allylation and crotylation of in situ generated N unsubstituted ketimines. PMID- 16358061 TI - Absolute configuration of amino alcohols by 1H-NMR. AB - A general NMR spectroscopy protocol for determination of absolute configuration of 1,2-amino alcohols, that allows differentiation of the four possible stereoisomers by analysis of the 1H NMR spectra of their bis-MPA derivatives, is described. PMID- 16358062 TI - Zipper-like properties of [poly(L-lysine)+ poly(L-glutamic acid)]beta-pleated molecular self-assembly. AB - Through the application of Molecular Force Spectroscopy, here we demonstrate nanomechanical properties of a beta-pleated, reversible, and pH-controllable molecular "zipper" formed by the spontaneous self-assembly of two sequenceless polypeptides. PMID- 16358063 TI - Small molecules as cross-linkers: fabrication of carbon nanotubes/thionine self assembled multilayers on amino functionalized surfaces. AB - Electroactive and photoactive thionine molecules have been selected as cross linkers to construct self-assembled multilayers containing carbon nanotubes (CNTs)via the alternate layer deposition technique. The resulting multilayer system can electrocatalyze the oxidation of NADH and offer potential applications in other fields such as biosenors and photovoltaic devices. PMID- 16358064 TI - Inhibition of dimethylarginine dimethylaminohydrolase (DDAH) and arginine deiminase (ADI) by pentafluorophenyl (PFP) sulfonates. AB - A range of pentafluorophenyl (PFP) sulfonate esters derived from the reaction of PFP vinyl sulfonate and various nitrones are shown to have significant inhibitory activity against the bacterial enzymes DDAH and ADI. PMID- 16358065 TI - DNA-intercalation on pyrene modified surface coatings. AB - Utilising the strong affinity between nucleic acids and an intercalating pyrene derivate, a novel efficient method for unspecific immobilisation of double stranded DNA on to solid support for applications in bioanalytic, biophysics and microbiology is presented. PMID- 16358066 TI - Pyrene-benzoylthiophene bichromophores as selective triplet photosensitizers. AB - Combination of the pyrene and benzoylthiophene units constitutes an interesting approach to design bichromophoric photosensitizers with increased intersystem crossing quantum yield and enhanced selectivity. The potential of this strategy has been illustrated in the present work by using a model photoisomerization reaction. PMID- 16358067 TI - Formation of ZnO hexagonal micro-pyramids: a successful control of the exposed polar surfaces with the assistance of an ionic liquid. AB - Wurtzite ZnO hexagonal micro-pyramids, with all exposed surfaces being polar +/- (0001) and {1011} planes, have been successfully synthesized using ionic liquids as solvents. PMID- 16358068 TI - [TcCl4(H2O)2] and [Cl3(H2O)2TcOTc(H2O)2Cl3]--two molecular intermediates of the hydrolysis of technetium(IV). AB - Two molecular intermediates of the hydrolysis of technetium tetrachloride, cis [TcCl4(H2O)2] and [Cl3(H2O)2TcOTc(H2O)2Cl3], were isolated and structurally characterised, suggesting that the hydrolytic degradation of technetium(IV) compounds occurs stepwise with the polymeric 'TcO2...nH2O' as a less defined final product. PMID- 16358069 TI - A novel ruthenium catalysed deracemisation of alcohols. AB - The deracemisation of alcohols has been achieved with 65-75% ee using a non selective ruthenium catalysed oxidation to give an intermediate ketone followed by an enantioselective hydrogenation. PMID- 16358070 TI - Quinine/selectfluor combination induced asymmetric semipinacol rearrangement of allylic alcohols: an effective and enantioselective approach to alpha-quaternary beta-fluoro aldehydes. AB - A quinine/Selectfluor combination inducing rearrangement reaction of allylic alcohols was discovered, which involved a moderate yield with good enantioselective construction of alpha-quaternary carbon center and beta-fluoro aldehyde under base condition. PMID- 16358071 TI - Total synthesis of aspercyclide C. AB - The first total synthesis of (+)-aspercyclide C (1) is reported using a kinetically controlled RCM reaction to form the 11-membered, unsaturated lactone ring of this bioactive diaryl ether macrolide. PMID- 16358072 TI - [Epidemiology of thyroid cancer in Brazil: pointing directions in the health policy of the country ]. PMID- 16358073 TI - [Growth hormone deficiency: up to where trust stimulation tests?]. PMID- 16358074 TI - [Interventions on the prevention of type 2 diabetes mellitus: is it feasible a population-based program in our country?]. AB - Considering the increasing prevalence of diabetes mellitus (DM) in underdeveloped countries as well as the simplicity of identifying individuals at high risk for such disease, implementation of intervention measures for its prevention is of great interest. Several studies have confirmed the benefits of lifestyle changes in preventing or postponing the progression from impaired glucose tolerance to DM. The review of these studies showed a 50% to 60% reduction in the incidence of DM by means of lifestyle modifications. Such results are better than those reported in studies in which pharmacological interventions were used with the same purpose. Despite the efficacy of lifestyle changes for the prevention of DM, compliance may represent a limitation to be implemented in communities. In our country, studies are necessary to assess the barriers for the implementation of a population-based program for the prevention of DM and other lifestyle related diseases in high-risk Brazilian subjects. PMID- 16358075 TI - [The role of the diet as a risk factor for the development and progression of diabetic nephropathy]. AB - Diabetic nephropathy (DN) is the leading cause of kidney disease in patients starting renal replacement therapy, and affects up to 40% of type 1 and type 2 diabetic patients. Diet seems to play an important role in the development of the disease. There are evidences supporting the concept that not only the amount but also the origin of dietary protein are associated with DN. Few studies analyzed the role of dietary lipids. A low-protein diet slows down the decline of renal function and ameliorates the DN prognosis and death in patients with type 1 diabetes with micro- and macroalbuminuria. Studies in type 2 diabetic patients are scanty but short-term studies suggest that this approach decreases albuminuria. However, the use of low-protein diet for long periods is compromised by poor compliance and its long-term safety is not firmly established. Enthusiastic results come up when comparing the effect of different sources of animal protein on renal function and lipid profile in patients with DN, which may represent an alternative strategy for low-protein diet on medical nutritional therapy in patients with DN and in cardiovascular risk factors and endothelial function. PMID- 16358076 TI - [Role of vitamin D in the neuro-muscular function]. AB - Through its action in the kidney, intestines, bone and parathyroid glands vitamin D is a major regulator of calcium homeostasis and for the development of a healthy skeleton. Moreover, receptors for this hormone are present in almost all body tissues and other actions which are not related to the mineral metabolism have been imputed to it. In the skeletal muscle cell, vitamin D acts through the classic mechanism of binding to a nuclear receptor and also by binding to a membrane receptor, carrying out actions that involve calcium transport, protein synthesis and kinetics of muscle contraction. Clinically, vitamin D deficiency, which is very common among the elderly, including the ones in our country, has been related to an increase in the incidence of falls, as well as the reduction of muscle strength and deterioration of body sway, evaluated by the oscillation of the body in the erect position. On the other hand, it has been demonstrated that supplementation of calcium associated to vitamin D in deficient elderly contributes to the improvement of these aspects of the neuro-muscular function. In this review, the mechanisms involved in the association between vitamin D and neuro-muscular function will be discussed, as well as the supplementation of vitamin D and calcium to prevent non-vertebral osteoporotic fractures under the perspective of the neuro-muscular effects. PMID- 16358077 TI - [Incidence and mortality from thyroid cancer in Brazil]. AB - In this study we evaluated the incidence and mortality due to thyroid cancer (TC) in Brazil using incidence data provided by seven Brazilian cancer registries and mortality data from the Brazilian Mortality Information System. Five-year age adjusted mortality rates were calculated over a 20-year period (1980-1999) for the country as a whole. We have calculated a 3-year age-adjusted incidence rate using data available since 1993. Age-adjusted mortality rates decreased from 0.22/100,000 to 0.28/100,000 (-21%) among males, and from 0.42/100,000 to 0.51/100,000 (-17%) among females. Among males, age-adjusted incidence rates varied from 0.7/100,000 in Belem to 3.0/100,000 in Sao Paulo. These cities also presented the lowest (0.8/100,000) and the highest (10.9/100,000) age-adjusted incidence rates among females. The downward tendency of mortality is probably explained by an improvement in diagnosis and treatment of TC over the study period, whereas geographical variations in incidence are probably related to availability of medical care resources in the different regions and the quality of cancer registers data. PMID- 16358078 TI - [Tolerance of the oral clonidine test in 180 patients: efficacy of the volemic expantion in controlling arterial hypotension]. AB - Clonidine stimulation test is widely used to evaluate growth hormone secretion. Side effects are somnolence (35%) and arterial hypotension (AH) (5%). The aims of this paper were to evaluate the tolerance to this test regarding blood pressure (BP) decrease, sedation and the efficacy of saline resuscitation to prevent AH. BP was measured at basal, 60 and 120 min. Sedation was determined by the Ramsay scale. Patients were divided into two groups: Group 1 (n = 80) received saline resuscitation only upon severe AH (drop of mean BP [MBP] > 20% from initial MBP) and/or postural hypotension; Group 2 (n = 100) received saline resuscitation from the beginning of the test. Both groups presented a significant MBP fall and 75% presented somnolence at 60 min. MBP drop did not correlate with either sedation or the clonidine dose. Group 1 presented more hypotension (59% x 28%) and greater MBP drop at 60 min. Only one patient had an asthma attack. We conclude that the hypotension effects caused by oral clonidine diminish with saline resuscitation since the beginning of the test. This test must have specialized medical support with strict BP evaluation and precocious intervention when needed. PMID- 16358079 TI - [Correlation between urinary iodine levels and TSH of umbilical cord from newborns of the University Hospital at the School of Medicine from Ribeirao Preto, Sao Paulo]. AB - The safest and most efficient way to diagnose congenital hypothyroidism (CH) is through screening programs using serum thyroid stimulating hormone (TSH). CH occurs in one in 1:2,300-1:5,000 births but higher rates are found in iodine deficient areas. Iodine was measured in the urine of 141 newborns (87 males and 54 females) from Ribeirao Preto to complement the screening program developed by the School of Medicine of Ribeirao Preto (FMRP-USP) and Federal University of Sao Paulo (UNIFESP). TSH values did not disclose any case of CH, although we have seen an elevated ratio (0.96%) of calls for retests. The iodine urinary levels ranged from 2.1 to 194 microg/l (mean 58.3+/-36.2 microg/l). No differences between the levels of urinary iodine and gender or gestational ages were observed. A negative correlation between urinary iodine and TSH of blood from umbilical cord was found (r= -0.20, p= 0.02). PMID- 16358080 TI - [Congenital transient hypothyroidism: characteristics of children identified at Newborn Screening Program of the State of Minas Gerais, Brazil]. AB - Early diagnosis and treatment of congenital hypothyroidism are the main goals of the neonatal screening programs. A retrospective analysis was carried out to assess the characteristics and etiologies of congenital transient hypothyroidism (CTH) in the Newborn Screening Program of the State of Minas Gerais, Brazil. To reach a diagnosis of CTH, altered screening and serum confirmatory T4 and TSH tests and normal serum free T4 and TSH at 4 and 8 weeks after the withdrawal of levothyroxine were used. We studied 21 children with CTH who represented 4.23% of the whole group with detected hypothyroidism and followed them up in the Screening Program. These patients received LT4 therapy for a variable period of one month to three years. Serum confirmatory TSH levels varied from 10.4 to 583.4 microUI/mL. Maternal antibodies, TRAB and anti-TPO, were associated to CTH in two children and iodine overload in one of them. We concluded that CTH is an important component in neonatal screening and hormonal treatment during the first months of the life is essential. The TSH initial levels are not relevant to determine whether the thyroid dysfunction is transient or permanent. PMID- 16358081 TI - [Body proportions in a group of Brazilian patients with Turner Syndrome]. AB - OBJECTIVE: The first Brazilian study aimed to evaluate body proportions in patients with Turner Syndrome (TS) with no growth hormone treatment. METHODS: A cross-sectional study with 50 TS patients (5 to 43 years) evaluating age, karyotype, pubertal development, height, sitting height, arm span, weight, BMI, head circumference, length of hand, foot and leg, waist to hip ratio, biacromial and biiliac diameters. The data were transformed in score z. A descriptive analysis was done and Mann-Whitney test and analysis of variance was applied. RESULTS: Twenty-eight patients were pubertal and 22 nonpubertal, and all of the absolute measurements were significantly increased in the former. There were no significant differences of the anthropometric findings according to the karyotype and puberty, in relation to score z data. The SD scores of all variables showed means above -2, except height and arm span in the nonpubertal patients and also sitting height and hand in the pubertal patients. The same result was observed when only the patients > 20 years were analyzed, being only weight, BMI, and biacromial and biiliac diameters significantly smaller in relation to the Danish data of Gravholt and Naeera from 1997. CONCLUSION: The retardation of growth in TS occurs mainly on the longitudinal axis, and the results showed in this study are similar to the Danish report. PMID- 16358082 TI - [Idiopathic central precocious puberty in girls in Bahia, Brazil]. AB - INTRODUCTION: Although the therapeutic effects of GnRH analogues (GnRHa) in central precocious puberty (CPP) have been established, clinical aspects may vary according to racial groups. OBJECTIVE AND METHODS: To describe the profile of 175 girls from Bahia, Brazil, treated with GnRHa for idiopathic CPP. RESULTS: The mulatto racial group predominated in the sample. The chronological age at puberty was 6.3 +/- 0.1 years. At diagnosis, height was 2.2 +/- 0.1 SD and body mass index (BMI) was 1.5 +/- 0.1 SD (> 2 SD in 32%), this one, inversely associated with age at onset of puberty (R = -0.20, p = 0.008). At treatment, chronological and bone ages were 8.4 +/- 0.1 years and 10.3 +/- 0.1 years, respectively. At the end of treatment (n = 52) age, BMI and height were 10.6 +/- 0.1 years, 1.6 +/- 0.2 SD and 1.9 +/- 0.2 SD, respectively. CONCLUSION: Clinical presentation and evolution were similar to other studies, independently of racial characteristics. In this sample, treatment with GnRHa was not associated to significant weight gain. PMID- 16358083 TI - [Association between serum markers for celiac and thyroid autoimmune diseases]. AB - Celiac disease (CD) is an autoimmune disease of the small bowel characterized by a strong genetic association with HLA - DQ2 and DQ8. Gluten is the etiological factor and the tissue enzyme transglutaminase (TGase) is its autoantigen. CD is associated with several autoimmune diseases such as type 1 diabetes, systemic lupus erythematous, rheumatoid arthritis, Sjogrens syndrome and autoimmune thyroid diseases. The aim of this study was to investigate the occurrence of serum IgA anti-endomysial and anti-human TGase antibodies in individuals with positive anti-thyroid antibody (ATA). The concordance between these two tests was also evaluated. Anti-endomysial antibodies were positive in 10 out of 456 (2.2%) and anti-human TGase were positive in 14 of 454 (3.1%) individuals with positive ATA. In control subjects they were positive in 1 of 197 (0.5%) and 2 of 198 (1%) for anti-endomysial and anti-human tissue TGase antibodies, respectively. The odds ratio (OR) for the anti-endomysial antibodies was 4.42 and for the anti human TGase 3.12 in individuals with ATA when compared with controls. An elevated concordance index (k= 0.84) was observed between anti-endomisyal antibodies and anti-human TGase. We conclude that the determination of anti-TGase antibodies is a good test for DC screening. PMID- 16358084 TI - [Evaluation of the knowledge of preventive measures for the diabetic foot in patients of Rio Branco, Acre]. AB - The diabetic foot is one of the main complications of diabetes mellitus (DM) of high socio-economic impact, characterized by foot lesions resulting from neuropathy, ischemia and infection. To verify the knowledge of preventive measures for the diabetic foot in a public health institution in Rio Branco, Acre, between August 2002 and February 2003 patients with type 2 DM (n= 109 patients, 71.6% female) were evaluated regarding their clinical-epidemiological history, demographic attitudes towards DM control and preventive care of the feet, before and after attending an informative educational program. There was a significant improvement of knowledge of preventive measures for the diabetic foot after the program (p< 0.0001), as well as for the attitudes towards DM control (p< 0.0001). We conclude that the population sample evaluated is unaware of preventive measures for the diabetic foot and of attitudes related to glycemic control. The utilization of educational programs, easily implemented in health centers, might reduce the rates of lower extremity amputation in patients with DM. PMID- 16358085 TI - [Association of gamma-glutamyl transferase and the metabolic syndrome in obese women]. AB - OBJECTIVE: To evaluate the correlation of gamma-glutamyl transferase (GGT) activity to the presence and number of abnormalities of the metabolic syndrome (MS). SUBJECT AND METHODS: We have studied 353 women consecutively admitted into the obesity outpatient unit, aged 39.0 +/- 11.4 years and body mass index 41.3 +/ 7.3 kg/m2. All patients were asymptomatic, had no history of alcohol abuse or liver disease. National Cholesterol Education Program--Adult Treatment Panel III criteria for diagnosing MS were applied. RESULTS: The frequency of the MS was 50.7% in the patients. The frequency of elevated GGT activity was higher in patients with MS than without it (32% versus 20%, p = 0.05). The average GGT concentration in patients with the MS was significantly higher than in non SM patients (52.8 +/- 42.3 U/L; Md: 40.0 versus 40.1 +/- 28.8 U/L; Md: 33.0, p = 0.0001). It was also higher in patients with each one abnormality of the MS, when compared to those without it. In patients with association of three abnormalities of the MS, the mean GGT activity was 51.0 +/- 39.2 U/L (Md: 36.0); in those with four, 50.8 +/- 50.4 U/L (Md: 40.5); and five, 64.8 +/- 32.3 U/L (Md: 61.0). There were positive correlations between GGT and glucose, triglycerides or blood pressure (systolic and diastolic). CONCLUSION: GGT activity was associated to the presence and the number of abnormalities of the MS, as well as to each one abnormality of the MS in obese women. PMID- 16358086 TI - [Accuracy, effect on insulin therapy and glycemic control and complications of the continuous glucose monitoring system in type 1 diabetic patients]. AB - To evaluate the efficacy, safety and complications of continuous glucose monitoring system (CGMS) in type 1 diabetic patients (DM1), we retrospectively studied 30 patients (25.8 +/- 12.2 years) submitted to 72 hs CGMS (Medtronic; Northridge, CA) and analyzed: mean self monitoring blood glucose (SMBG) and mean CGMS sensors glycemic value; correlation coefficient (%), median absolute percent difference (MAD%), number of sensor reading, glycemic excursions (CGMS vs. SMBG), complications (trauma, local infection, disconnection) and therapeutic management after CGMS. A1c levels were measured 1 month before and 3 months after the study. Mean capillary glucose values were 186.5 +/- 43.3 mg/dl vs. 179.7 +/- 48.1 mg/dl by CGMS sensor, with significant correlation (p = 0.001). An average of 772.4 +/- 254.1 (VR > 680) glucose measurements was recorded for each patient, with 68.7 +/ 19.8 hs of exam. Correlation coefficient was 0.86 +/- 0.21 (VR > 0.79). Median absolute percent difference between sensor and glucometer values was 13.9 +/- 4.7% (VR < 28%). The CGMS was significant more efficient in detection of glycemic excursion related to capillary glycemia (p = 0.009). This data showed important decreased level of A1c in this population 3 months after the CGMS with statistical significance (p = 0.018). No complications were registered in 96.7% of patients. No trauma, local infection or bleeding were registered. The insulin therapeutic regimen was adjusted in 100% of patients. The CGMS showed to be a very safety method, well tolerated, with high accuracy in glycemic values and low complications rate. This method has to be more stimulated by physicians and patients. PMID- 16358087 TI - Efficacy of continuous glucose monitoring system to detect unrecognized hypoglycemia in children and adolescents with type 1 diabetes. AB - This retrospective study assessed 17 DM1 pediatric patients (15.76 +/- 4.5 years) submitted to 72 h continuous glucose monitoring system (CGMS) (Medtronic, CA). The aim of this study was to evaluate the accuracy of CGMS in children and adolescents with type 1 diabetes mellitus (DM1) and the efficacy of this method to detect unrecognized hypoglycemia in this population. It were analyzed capillary glycemia (CG) and CGMS sensors value; glycemic excursions; postprandial hyperglycemia; unrecognized hypoglycemia; complications and therapeutic management after CGMS. A1c levels were measured at the start and after 3 months of the study. Correlation coefficient during hypo, hyper, and normoglycemia and sensitivity/specificity was determined. The mean CG values were 213.8 +/- 63.4 mg/dl vs. 209.7 +/- 52.5 mg/dl by sensor, with statistical significance by Pearson's correlation (p < 0.001). There was no difference between CGMS and CG value in order to detect glycemic excursions (p = 0.32). The postprandial hyperglycemia and unrecognized hypoglycemia was detected in 66.7% and 56.2% of this patients, respectively. The correlation coefficient during hypoglycemia presented no statistical significance by Pearson's correlation (p = 0.29) vs. during hyperglycemia (p = 0.001). The CGMS sensor presented low sensitivity (63.3%) to detect hypoglycemia. This data showed important decreased level of A1c in this population 3 months after CGMS with statistical significance (p = 0.03). The CGMS showed to be a very safe method, well tolerated, with high accuracy in glycemic values and low complications rate. This results suggest that CGMS is a good method to identify postprandial hyperglycemia, to improve metabolic changes in therapeutics with significant impact in A1c of diabetic pediatric patients. This data confirmed the low sensitivity of CGMS to detect unrecognized hypoglycemia in pediatric DM1 patients. PMID- 16358088 TI - [Cost-effectiveness of the clinical treatment of Grave's disease in a public University Hospital: a retrospective analysis and prospective projection for a therapeutic approach]. AB - The aim of the present study was to evaluate a new proposal for increasing compliance to the clinical management of patients with Graves' disease (GD) in a large and public University Hospital. The patients were carefully selected (no previous GD treatment, goiter volume less than 6 mL must be living in the metro area of Sao Paulo), received medication at no cost, were contacted frequently by the social worker and alerted for the date of consultation and only referred to a single endocrinologist during all phases of treatment. We recruited 229 patients with GD that were initially treated with methimazole (MMI--60 mg q.d) in a single daily dose followed by a combination of MMI (20 mg) plus L-T4 (100 microg) daily for 24 months. Only 83 patients (36.2%) completed the protocol and were subdivided in: Group 1 (n= 34) that were in remission for 3 years after discontinuation of the MMI and Group 2 (n= 49) that presented recurrence of GD between 2 and 36 months without MMI. Predictive factors associated with remission were: decrease of the glandular volume, serum TG< 40 ng/mL and normal TRAb values. We concluded that in spite of a careful protocol planned to increase compliance, more than 60% of patients with GD did not complete the therapeutic trial and were referred for radioiodine treatment. The solution for this low therapeutic success for GD should be the possible identification of factors that would indicate patients that are not inclined to follow a long period of clinical therapy. PMID- 16358089 TI - [Online continuing medical education in endocrinology and metabology]. AB - The World Wide Web, also known as the Internet, is an important agent of spreading human knowledge. Medical education through the web without formal presence at the classroom is a modern use of this valuable resource, characterizing the online continuous medical education (eCME). The search of videoconferences, monographs, guidelines and update courses in endocrinology and metabolism at the Internet through search mechanisms is an extremely slow and hard task due to the huge amount of sites resulting from this process. This revision aims to make this quest easier, by presenting in a rational order, according to the subspecialty, glandular organ and language, the most important web pages that deal with this subject, obtained in a previous research promoted by the author. PMID- 16358090 TI - [Mario Philomeno Caleiro Lima: the trajectory in the construction of the endocrinology]. PMID- 16358091 TI - [Autoimmune hypothyroidism nonresponsive to high doses of levothyroxine and severe hypocalcemia]. AB - The factors that make difficult the normalization of TSH in hypothyroidism need special attention because some patients on thyroxine replacement do not maintain a normal TSH. We report a 50 year-old woman with autoimmune hypothyroidism of difficult compensation, associated with anemia, hypocalcemia with a previous episode of tetany, hypomagnesemia, psychologic alterations and important weight loss. After compensation of the hypothyroidism with doses of L-thyroxine as high as 325 microg/day, the hypothesis of a malabsorptive syndrome was raised. Celiac disease was confirmed by elevated serum antigliadin antibody. A gluten-free diet was instituted which improved the symptoms associated with malabsorption and reduced the L-thyroxine requirement to 125 microg/day. Because several studies have shown an association of both diseases, a routine screening for celiac disease has been widely proposed in patients with autoimmune thyroid disease. PMID- 16358092 TI - [High doses statins administration causing rhabdomyolysis: case report]. AB - Rhabdomyolysis has been the theme in medical literature for the last fifty years. In these last decades, with statins being used in primary and secondary cardiovascular prevention events, this theme returns and statins are now pointed as the trigger to this almost always fatal complication. Rhabdomyolysis due to statins administration occurs mainly in association with other drugs. Our case reports on a patient with fatal statin-induced rhabdomyolysis whose medical history included diffuse atherosclerotic disease. PMID- 16358093 TI - The "common soil" theory: coronary disease, diabetes and inflammation. PMID- 16358094 TI - Psychoactive drug advertising: a comparison of technical information from three countries: Brazil, United States and United Kingdom. AB - CONTEXT AND OBJECTIVE: Studies carried out in the 1970s and 1980s showed that there were country-dependent disparities in the information given for the same drug in medical advertisements. National and international regulations have been published to do away with such disparities and to foster the rational use of drugs. The purpose of this study was to compare the information contained in psychoactive drug advertisements published in psychiatric journals in Brazil, the United States and the United Kingdom, before and subsequent to the publication of the United States Export Act, in 1986, the WHO criteria, in 1988, and the Brazilian Sanitary Surveillance Agency Resolution no. 102, in 2000. TYPE OF STUDY AND SETTING: Content analysis, at Centro Brasileiro de Informacoes sobre Drogas Psicotropicas (Cebrid). METHODS: We gathered advertisements from Brazilian, American and British psychiatry periodicals published before and after each ruling. We analyzed a total of twenty-four Brazilian advertisements that were for the same psychoactive drugs as advertised in American and/or British publications from the same period. RESULTS: We observed that Brazilian advertisements omitted information on usage restrictions, such as contraindications, adverse reactions, interactions, warnings and precautions, and that such information was present in American and British advertisements. CONCLUSIONS: The data suggest that disparities in the information given for the same drug still persist. The information depends on the country in which each drug is marketed. The legislation is insufficient for eradicating such disparities. PMID- 16358095 TI - Salivary cortisol as an indicator of adrenocortical function in healthy infants, using massage therapy. AB - CONTEXT AND OBJECTIVE: The evaluation of adrenocortical function with the use of therapeutic massage has been little studied in Brazil. The purpose of this study was to evaluate the salivary cortisol levels before and after Shantala massage therapy on healthy infants. DESIGN AND SETTING: Prospective case series, in a public nursery, in Sao Paulo. METHODS: Saliva was obtained from 11 infants at the times of 8:00-9:00 a.m. and 4:00-5:00 p.m. in a nursery and 9:00-10:00 p.m. at home. They received a 15-minute therapeutic massage on two consecutive days, and saliva was collected before and after the massage. The procedure was repeated after a one-week interval. Cortisol values (intra-assay < 5%; inter-assay < 10%) at different times of the day were compared by ANOVA. RESULTS: The mean cortisol values (nmol/l +/- SD) on the first day were: morning (M) = 14.1 +/- 5.7, afternoon (A) = 8.3 +/- 2.7, night (N) = 3.3 +/- 1.1; after two consecutive days of therapeutic massage: M = 22.3 +/- 13.5, A = 13.4 +/- 6.0, N = 5.8 +/- 3.5; after a one-week interval: M = 15.8 +/- 7.7, A = 14.3 +/- 7.7, N = 3.4 +/- 2.0. CONCLUSION: There was a modification in the salivary cortisol values following massage, thus reflecting possible adaptation of the hypothalamic-pituitary adrenal axis. PMID- 16358096 TI - Continuous infusion of amphotericin B: preliminary experience at Faculdade de Medicina da Fundacao ABC. AB - CONTEXT AND OBJECTIVE: Intravenous amphotericin B deoxycholate (AmB-D) infusions, usually given over 4 hours, frequently induce nephrotoxicity and undesirable infusion-related side effects such as rigors and chills. There is evidence in the literature that the use of AmB-D in the form of continuous 24-hour infusion is less toxic than the usual four-hour infusion of this drug. Our objective was to evaluate the efficacy and safety of continuous infusion of AmB-D for the treatment of persistent fever in neutropenic patients with hematological malignancies after chemotherapy. DESIGN AND SETTING: Observational retrospective analysis of our experience with continuous infusion of AmB-D, at Faculdade de Medicina da Fundacao ABC and Hospital Estadual Mario Covas in Santo Andre. METHODS: From October 2003 to May 2004, 12 patients with hematological malignancies and chemotherapy-induced neutropenia received 13 cycles of continuous infusion of AmB-D. RESULTS: The median dose of AmB-D was 0.84 mg/kg/day (0.33 to 2.30 mg/kg/day). Concomitant use of nephrotoxic medications occurred in 92% of the cycles. Nephrotoxicity occurred in 30.76% of the cycles, hypokalemia in 16.67%, hepatotoxicity in 30% and adverse infusion-related events in 23%. All patients survived for at least seven days after starting continuous infusion of AmB-D, and clinical resolution occurred in 76% of the cycles. CONCLUSIONS: Continuous infusion of AmB-D can be used in our Institution as an alternative to the more toxic four-hour infusion of AmB-D and possibly also as an alternative to the more expensive liposomal formulations of the drug. PMID- 16358097 TI - Motivation to change drinking behavior: the differences between alcohol users from an outpatient gastroenterology clinic and a specialist alcohol treatment service. AB - CONTEXT AND OBJECTIVE: For some patients who have developed significant alcohol related physical disease, total abstinence from alcohol may offer the best chance of survival. The aim of this study was to investigate motivation for treatment in two groups of alcohol users: outpatients from the gastroenterology clinic and outpatients from the specialist alcohol treatment service. DESIGN AND SETTING: Cross-sectional study, at a federally funded public teaching hospital. METHODS: The sample studied was 151 outpatients from the gastroenterology clinic and 175 from the specialist alcohol treatment service. The interview was conducted in the outpatient clinics at the first appointment, and consisted of demographic questions and scales for measuring quality of life, alcohol dependence, pattern of alcohol, motivation for treatment and consequences of alcohol consumption. RESULTS: The results suggested that outpatients from the gastroenterology clinic were less dependent on alcohol, had suffered fewer consequences from alcohol and had fewer emotional and mental health problems than did the outpatients from the alcohol treatment service. In relation to their stages of change, the gastroenterology outpatients presented high precontemplation scores at the beginning of treatment while outpatients of alcohol treatment service showed higher scores in contemplation, action and maintenance. CONCLUSION: The medical treatment may be a reason for the temporary alcohol abstinence behavior among the gastroenterology outpatients. PMID- 16358098 TI - Implementing a clinical assessment protocol for sensory and skeletal function in diabetic neuropathy patients at a university hospital in Brazil. AB - CONTEXT AND OBJECTIVE: Physiotherapy can contribute towards recovering or preventing physical and sensory alterations in diabetic neuropathy patients. Our objective was to create and apply a protocol for functional assessment of diabetic neuropathy patients' lower limbs, to guide future physiotherapy. DESIGN AND SETTING: Clinical study at the University Hospital and teaching/research center of Universidade de Sao Paulo. METHODS: An intentional sample of diabetic neuropathy patients was utilized. The protocol was divided into: (1) preliminary investigation with identification of relevant clinical diabetes and neuropathy characteristics; (2) thermal, tactile and proprioceptive sensitivity tests on the feet; (3) evaluations of muscle function, range of motion, lower limb function, foot anthropometry. RESULTS: The patients' mean age was 57 years, and they had had the diagnosis for 13 years on average. Distal numbness and tingling/prickling were present in 62% and 67%, respectively. There were tactile sensitivity alterations above the heel in 50%, with thermal sensitivity in 40% to 60%. The worst muscle function test responses were at the triceps surae and foot intrinsic muscles. Longitudinal plantar arches were lowered in 50%. Decreased thermal and tactile sensitivity of the heels was found. There was a general reduction in range of motion. CONCLUSIONS: The results provided detailed characterization of the patients. This protocol may be easily applied in healthcare services, since it requires little equipment, at low cost, and it is well understood by patients. PMID- 16358099 TI - Prevalence of sexual problems and related help-seeking behaviors among mature adults in Brazil: data from the global study of sexual attitudes and behaviors. AB - CONTEXT AND OBJECTIVE: Relatively little is known about the usual frequency of sexual activity and how older individuals cope with sexual problems. The objective was to study sexual activity, prevalence of sexual problems and related help-seeking behaviors among middle-aged and older men and women in Brazil. DESIGN AND SETTING: Population survey, by Fundacao Oswaldo Cruz. METHODS: Interviews were held with 1,199 Brazilians aged 40-80 years (471 men and 728 women). The standardized questionnaire investigated demographics, general health, sexual behavior, attitudes and beliefs. RESULTS: Overall, 92.6% of men and 58.3% of women had had sexual intercourse during the preceding year. More than half of the men and women had done so more than once a week. Early ejaculation (30.3%) was the commonest male sexual problem, followed by inability to reach orgasm (14.0%), erectile difficulties (13.1%) and lack of sexual interest (11.2%). For women, the commonest sexual problems were lubrication difficulties (23.4%) and lack of sexual interest (22.7%). Depression was a significant correlate of sexual problems, for men and women. More women than men had sought help for sexual problem(s) from a healthcare professional. CONCLUSIONS: The findings highlight the importance of encouraging greater use of available healthcare services, including consultation with a medical doctor regarding sexual health. This should not only enable men and women to maintain satisfactory sexual function well into their later years, but may also result in overall improvement in the quality of healthcare. PMID- 16358100 TI - Authorship: an ethical dilemma of science. AB - CONTEXT AND OBJECTIVE: The scientific and technological progress that has taken place since the 1960s has brought an ever-growing volume of scientific research, and inflation in co-authorship. Over this period, it has been observed that an increasing number of publications have listed authors or co-authors whose participation in the published research was minimal or even nonexistent. The objective of this work was to analyze reports in the literature regarding misconduct in authorship: its types, chief causes, consequences and ethical guidelines; and to outline proposals for greater ethical commitment in scientific publication. DESIGN AND SETTING: Narrative review undertaken at Faculdade de Medicina de Itajuba, Minas Gerais, Brazil. METHODS: Analysis of publications about authorship using the Medline, Lilacs and SciELO databases. RESULTS AND CONCLUSIONS: Frequent types of misconduct were gift authorship and divided and redundant publications. The chief causes of these practices seem to be the pressure exerted by academia and the desire for social and professional development. Such factors have brought an increase in unethical behavior. This bias in science continues despite the criteria defined by the International Committee of Medical Journal Editors, the Vancouver group. RECOMMENDATIONS: Various actions are proposed for educational institutions, research development agencies, regulatory agencies and professional associations. The aim is to establish an evaluation policy that gives primacy to the quality of publications and sets ethical principles for scientific research. PMID- 16358101 TI - Neutropenia, agranulocytosis and dipyrone. AB - CONTEXT: Neutropenia and agranulocytosis may be defined as granulocyte counts of less than 1,500/mm3 and 500/mm3, respectively. Agranulocytosis is a rare and serious disease often caused by drugs. Its mortality rate is around 10%. The most common manifestations are infections such as tonsillitis, pharyngitis, stomatitis or pneumonia. Although dipyrone is one of the drugs known to be associated with agranulocytosis, the strength of the association has been a matter of much debate. Moreover, alternative analgesic and antipyretic agents are not devoid of serious side effects. CONCLUSIONS: It is therefore necessary to establish the incidence of agranulocytosis in Latin America and the role of dipyrone. The ongoing LATIN Study is a multicenter international case-control study that will provide answers for these questions. PMID- 16358102 TI - Cellular angiofibroma of the vulva: case report with clinicopathological and immunohistochemistry study. AB - CONTEXT: Cellular angiofibroma of the vulva is a rare tumor that was first described in 1997. It occurs in middle-aged women (average age: 47 years), has small size (< 3 cm) and well-circumscribed margins. CASE REPORT: We describe a case in a 51-year-old woman whose preoperative diagnosis was confounded with Bartholin's glandular cyst. The neoplasia was well delimited and made up of three characteristic components: fusiform cells forming small fascicles, numerous blood vessels and adipose tissue interspersed between the fusiform cells. The stroma cells were positive for vimentin and negative for CD34, protein S-100, actin and desmin. The differential diagnoses for this tumor include aggressive angiomyxoma, angiomyofibroblastoma, lipoma of fusiform cells, solitary fibrous tumor, perineurioma and leiomyoma. PMID- 16358103 TI - Fibroadenoma in axillary supernumerary breast: case report. AB - CONTEXT: Supernumerary breast tissue may be affected by the same diseases and alterations that compromise topical breast tissue. Nevertheless, reports of fibroadenoma in supernumerary breast tissue in the axillae are rare. OBJECTIVE: To describe a case of fibroadenoma in an axillary supernumerary breast. DESIGN: Case report. CASE REPORT: A 39-year-old woman was referred to the gynecology and obstetrics outpatient clinic at Hospital Estadual Sumare, complaining of bilateral axillary masses. The patient reported cosmetic problems and local pain and discomfort. On physical examination, alterations compatible with bilateral axillary accessory breasts, without palpable nodules, were observed. Supplementary examinations (mammography and ultrasonography) revealed a 1.1 cm mass in the right axillary breast. The patient underwent resection of the supernumerary breasts and histopathological examination revealed fibroadenoma of the right axillary breast tissue. PMID- 16358104 TI - Are we ready for endophenotypes in attention deficit hyperactivity disorder? PMID- 16358105 TI - Can we prevent Alzheimer's disease? PMID- 16358107 TI - Spontaneous harm reduction: a barrier for substance-dependent individuals seeking treatment? AB - OBJECTIVE: Greater information regarding motivations and treatment barriers faced by substance-dependent individuals has clinical and public health implications. This study aimed to formulate hypotheses regarding psychological, social and family variables that can be constructed as motivations or subjective barriers for the early seeking of formal treatment. METHODS: A qualitative study was conducted in an intentional sample (selected through saturation and variety of types) of 13 substance-dependent individuals who sought treatment. In-depth, semi structured interviews were conducted using open questions, and the transcribed data were subjected to qualitative analysis. RESULTS: Four types of spontaneous harm reduction measures were identified, according to the subjective logic of each participant: having some periods at rest (not using and recovering from adverse effects); caretaking by close acquaintances (relatives, partners, drug dealers and alcoholic beverage sellers); selectivity regarding substance source, type and means of administration; establishing "healthy" limits of ingestion. CONCLUSIONS: The measures identified might represent barriers to the early seeking of treatment but might also represent spontaneous learning of abilities beneficial to future treatment. Health care professionals should take into consideration their existence and should address them in clinical settings. Issues representative of the formulated categories should be presented in structured questionnaires used in future quantitative studies of barriers to treatment in this population. PMID- 16358106 TI - Depression, anxiety and quality of life scores in seniors after an endurance exercise program. AB - OBJECTIVE: Mood disorders are a frequent problem in old age, and their symptoms constitute an important public health issue. These alterations affect the quality of life mainly by restricting social life. The participation in a regular exercise program is an effective way of reducing or preventing the functional decline associated with aging. The aim of the present study was to examine the effects of fitness-endurance activity (at the intensity of Ventilatory Threshold 1 (VT-1)) in depression, anxiety and quality of life scores in seniors. METHODS: The study involved 46 sedentary seniors aged 60-75 (66.97 +/- 4.80) who were randomly allocated to two groups: 1) Control group, which was neither asked to vary their everyday activities nor to join a regular physical fitness program; and 2) Experimental group, whose members took part in an aerobic fitness program consisting of ergometer cycle sessions 3 times a week on alternate days for six months working at a heart rate corresponding to ventilatory threshold (VT-1) intensity. Subjects were submitted to a basal evaluation using the geriatric depression screening scale--GDS, STAI trait/state (anxiety scale) and SF-36 (quality of life scale). RESULTS: Comparing the groups after the study period, we found a significant decrease in depressive and anxiety scores and an improvement in the quality of life in the experimental group, but no significant changes in the control group. CONCLUSION: The data suggest that an aerobic exercise program at VT-1 intensity suffices to promote favorable modifications in depressive and anxiety scores to improve the quality of life in seniors. PMID- 16358108 TI - Method for evaluating subjective states of awareness that accompany recognition: adaptation for use in Portuguese-speaking patients with schizophrenia. AB - OBJECTIVE: Memory is composed of several systems and processes, and recognition can be accompanied by two subjective states of consciousness: autonoetic awareness, which characterizes conscious recollection; and noetic (or semantic) awareness, related to feelings of familiarity. The objective of this study was to describe the adaptation to Portuguese of an experiential procedure for investigation of states of awareness that accompany recognition. METHODS: Development of the material (word list and instructions permitting manipulation of the level of information processing); translation/adaptation of the original instructions to Portuguese; and application of the procedure in healthy volunteers and patients with schizophrenia. Manipulation of the level of processing consisted in requiring, during the learning phase, that subjects form a phrase or count the number of letters of the words presented. The level of processing, documented in healthy volunteers, should be expressed by greater conscious recollection of words used to form phrases than of words for which letters were counted. In addition, there should be no change in the proportion of recognition based on feelings of familiarity. RESULTS: The procedure was first applied in six healthy volunteers, in whom the processing level effect was clearly reproducible. Subsequently, it was used in patients with schizophrenia, who understood and followed the instructions perfectly. There was also a clear processing level effect in the patient group. CONCLUSION: The Portuguese version of this method can be used in our milieu, even in patients with schizophrenia, allowing the study of memory alterations accompanying this mental disorder. PMID- 16358109 TI - Clinical profiles of serious suicide attempters consecutively admitted to a university-based hospital: a cluster analysis study. AB - OBJECTIVE: To verify the presence of different groups of medically serious suicide attempters who had more clinical or surgical seriousness and required admission to a general hospital. METHODS: 121 patients admitted consecutively were assessed. A questionnaire containing items on the patient characteristics and psychometric scales to assess the suicidal intent and lethality were used. A cluster analysis was performed using the K-means method. RESULTS: Three groups were identified: 1) 43 subjects (mostly female) characterized by self-poisoning with medication and low suicidal intent, with highly impulsive suicide attempts; 2) 53 subjects (mostly males) who ingested pesticides and presented both moderate degrees of lethality and suicidal intent; 3) 17 subjects (predominantly males) who used more violent methods and presented high levels of lethality and suicidal intent. CONCLUSIONS: Grouped data of these inpatients could be misleading for follow-up research purposes as our findings indicate that there are relatively distinct clinical profiles among suicide attempters admitted to a general hospital. PMID- 16358110 TI - Relationship between binge-eating episodes and self-perception of body weight in a nonclinical sample of five Brazilian cities. AB - OBJECTIVE: To investigate the relationship between binge-eating episodes and a perception that body weight is above the ideal in a sample of customers interviewed at shopping malls in five Brazilian cities. METHODS: In 1999, data were collected over the course of one week (Monday-Friday only) at the largest shopping malls in the cities of Porto Alegre, Salvador, Fortaleza, Goiania and Curitiba (two malls per city). A total of 2855 participants (917 men and 1938 women) were interviewed. Weight and height measurements were standardized. Binge eating episodes were identified using a questionnaire including the following questions based on DSM-IV diagnostic criteria: "Have you ever eaten, in a period of two hours or less, an amount of food greater than that most people would eat?" and "If the answer was "yes", did you, during these episodes, feel unable to stop eating or to control how much you were eating?". RESULTS: The prevalence of binge eating episodes was higher among overweight subjects (15.6%) compared with normal weight subjects (9.9%) (p = 0.0001) and, among subjects who perceived their body weight to be above the ideal (men: 13.9%; women: 15.1%) compared with those who perceived their body weight to be ideal or below the ideal (men: 8%; women: 7%) (p < 0.0001). In the multivariate analysis adjusted for body mass index and demographic variables, binge-eating episodes were correlated with the perception that body weight is above the ideal only among women (OR: 1.8; 95% CI: 1.2-2.5). CONCLUSIONS: These findings suggest that the perception that body weight is above the ideal, regardless of overweight status, is associated with binge-eating episodes in women. PMID- 16358111 TI - Validity of Autism Behavior Checklist (ABC): preliminary study. AB - OBJECTIVE: To examine the concurrent and criterion validity of the Autism Behavior Checklist (ABC). METHODS: Three groups, comprising 38 mothers of children previously diagnosed with autism (DSM IV-TR, 2002), 43 mothers of children with language disorders other than autism, and 52 mothers of children who had no linguistic or behavioral complaints, were interviewed. In order to minimize the effect of maternal level of education, the questionnaire was completed by the researcher. To determine the concurrent validation, ANOVA and discriminant analysis were used. The ROC curve was used to establish the cutoff score of the sample and to examine the criterion validity. RESULTS: The mean total score was significantly higher in the group of mothers of autistic children than in the other groups. The ABC correctly identified 81.6% of the autistic children. The ROC curve cutoff score was 49, and the sensitivity was 92.1%, higher than the 57.89% found when a cutoff score of 68 was used. The specificity was 92.6%, similar to the 94.73% obtained with a cutoff score of 68. CONCLUSIONS: The ABC shows promise as an instrument for identifying children with autistic disorders, both in clinical and educational contexts, especially when a cutoff score of 49 is used. PMID- 16358112 TI - Factors associated with the successful aging of the socially-active elderly in the metropolitan region of Porto Alegre. AB - OBJECTIVE: To identify the factors associated with aging of the socially-active elderly in the metropolitan region of Porto Alegre. METHODS: A cross-sectional study was conducted involving a sample of 400 elderly subjects. Through a conglomerate analysis using the combined results of the Flanagan Scale and the quality of life assessment instrument developed by the World Health Organization (WHOQOL-100), the subjects classified as presenting a higher quality of life were considered to have aged successfully. RESULTS: After having been adjusted for the variables gender, age, level of depression, despair, perceived health status, meaning that personal beliefs give to life, satisfaction with social activities and relations, the following factors maintained their independent and meaningful effect on successful aging: family relations and friendships (OR = 4.93; 95% CI: 2.83-8.60), health and perceived well being (OR = 1.66; 95% CI: 1.07-2.59), functional capacity (OR = 23.70; 95% CI: 10.98-51.06) and psychosocial support (OR = 7.36; 95% CI: 4.00-13.54). CONCLUSIONS: Independence in performing daily life activities and autonomy, as well as satisfaction with family relations and friendships, were independent predictive factors of successful aging for both men and women. For women in particular, there were various predictive factors: material comfort; feeling physically well; body image and appearance; self esteem; positive feelings; interpersonal relationships; social support; participation in recreational activities; sexuality; spirituality; and beliefs. The elderly maintain a relationship with themselves, with others and with their beliefs. Family is the main social support system. PMID- 16358113 TI - The social adjustment of people with severe mental illness in Sao Paulo, Brazil. AB - OBJECTIVE: To investigate the social adjustment of individuals with severe mental illness living in the community in a large urban center of a developing country, and the characteristics associated with poor social functioning. METHOD: A cross sectional study was performed in the city of Sao Paulo. Eligible subjects were residents of a defined geographic area, aged between 18 and 65, with a diagnosis of functional psychosis who had had contact with any public psychiatric service during a defined period. Structured assessments were used to obtain information on social-demographic characteristics, diagnosis (ICD-10), psychiatric symptoms (PANSS), and social adjustment (DAS). RESULTS: One hundred and eighty-eight subjects were included, of whom, 120 (63.8%) had some degree of impairment in social functioning. The most frequently affected areas of social functioning were work performance and sexual role. Twenty-four patients (12.8%) showed poor or very poor social adjustment in the month prior to the interview. Negative symptoms, number of previous admissions and general symptoms showed statistically significant associations with global social adjustment scores. CONCLUSIONS: The proportion of patients showing any degree of impairment in social adjustment was as high as in more developed societies. In order to successfully implement the new mental health policy in Brazil, better provision of community-based mental health services for those with severe mental illnesses is needed. PMID- 16358114 TI - Nursing personnel attitudes towards suicide: the development of a measure scale. AB - OBJECTIVE: To describe the construction of the Suicide Behavior Attitude Questionnaire (SBAQ) which measures attitudes of nursing personnel towards suicide, and verify attitude differences among these professionals. METHODS: The Suicide Behavior Attitude Questionnaire comprises 21 visual analogue scale items (beliefs, feelings and reactions on suicidal patients) selected from a pool of attitude statements generated by focal groups and experts' judgement. The questionnaire was completed by 317 nursing professionals who worked in a teaching hospital. Factor analysis and internal consistency were calculated. RESULTS: Three interpretable factors were extracted, accounting jointly for 40% of the total variance: Feelings when caring for the patient, Professional Capacity and Right to Suicide, comprising 7, 4 and 5 items, respectively. The Cronbach's alpha coefficients were 0.7, 0.6 and 0.5, respectively. Greater Professional Capacity was reported by nursing assistants and those who had already took care of suicidal patients. The belief that a person does not have the right to commit suicide was stronger among older professionals, those who had never taken care of suicidal patients, those who had a family history of suicide, those who were Protestants and that used to go more frequently to church services. CONCLUSIONS: The Suicide Behavior Attitude Questionnaire proved to be user-friendly and quite a simple instrument to assess attitude towards suicide among nursing personnel. PMID- 16358115 TI - Validation of the Portuguese version of the Questionnaire on Eating and Weight Patterns-Revised (QEWP-R) for the screening of binge eating disorder. AB - OBJECTIVE: The present paper describes the validation of the Questionnaire on Eating and Weight Patterns-Revised (QEWP-R) designed for the diagnosis of binge eating disorder (BED) and sub-clinical binge eating. METHODS: 89 overweight women seeking treatment for binge eating and/or obesity were assessed with the Portuguese version of the Questionnaire of Eating and Weight Patterns and were, subsequently, interviewed with the eating disorders module of the Structured Clinical Interview for DSM-IV (SCID-I/P). Rates of binge eating disorder and sub clinical cases of binge eating obtained with the Questionnaire on Eating and Weight Patterns-Revised were then compared to those obtained with the Structured Clinical Interview for DSM-IV. RESULTS: In the identification of binge eating, irrespective of the presence of all criteria for binge eating disorder the QEWP-R Questionnaire on Eating and Weight Patterns-Revised yielded a sensitivity value of 0.88, a specificity value of 0.63 and a positive predictive value of 0.825. Rates for the identification of the full syndrome of binge eating disorder were: sensitivity value of 0.548, a specificity value of 0.8 and a positive predictive value of 0.793. CONCLUSIONS: The Questionnaire on Eating and Weight Patterns Revised can be useful in a first-step screening procedure to identify probable cases of binge eating. It can be useful as a screening tool and as a first step of clinical assessment of patients seeking treatment for binge eating and/or obesity. PMID- 16358116 TI - [Epidemiologic aspects and relevance of mental disorders associated with epilepsy]. AB - Epilepsy is the most common serious neurological disorder. Worldwide, around 50 million people have epilepsy, 40 million of which in developing countries. Though epilepsy is predominantly treatable, most patients in these countries remain without treatment. It is likely that one of the main reasons is the stigma associated with epilepsy. This problem increases when mental disorders are associated with epilepsy, a fairly frequent occurrence. Patients with epilepsy and mental disorders are exposed to a "double stigma", which frequently leads to complete absence of treatment. It is probable that, particularly in developing countries, psychiatrists will be involved with the treatment of people with epilepsy. Our purpose was to review the epidemiology of mental disorders associated with epilepsy, its treatment gap and impact upon quality of life of the patients. PMID- 16358117 TI - [Childhood anxiety and assessment instruments: a systematic review]. AB - OBJECTIVE: To make a survey of the principal anxiety instruments available for children, when they appeared, the type of methods used, verifying which countries have the highest number of children anxiety instruments and also how is the Brazilian reality on this matter. METHODS: A systematic review on electronic databases--Psychoinfo (1940--May 2002), Psyclit (1887--May 2002), Medline (1966- May 2002) e Eric (1966--May 2002). Information collected on personal communications and books chapters. The inclusion criterion utilized: studies realized with children, where the anxiety were evaluated with some psychometric instrument. The exclusion criterion: adolescents, adults and animal research articles, articles utilizing projective instruments or studies without any reference of assessment instruments. The indexed articles found in 2 or more databases were considered only once. RESULTS: This review pointed out 1911 studies used at least one assessment instrument to identify the presence of anxiety and 118 instruments were used for this purpose. CONCLUSIONS: There are a large number of children anxiety instruments available, however, in Brazil, the only one available in the market isn't updated and the modern ones are found only in clinic academic centers. PMID- 16358118 TI - Reviving the old sermon of medicine with the placebo effect. AB - OBJECTIVE: The message of the importance of a caring doctor-patient relationship is now like an old sermon which does not impact anyone's mind or action. Observing the healing practice of the old time physicians, who valued their attitudes and relationship with their patients more than the actual interventions, this paper reviews the literature on their main therapeutic device -the placebo effect--as a novel way of delivering this old sermon of medicine to contemporary doctors. DISCUSSION: There are countless historical and contemporary examples of the impressive placebo effect and although contested by some, it seems real and significant. The classic conditioning theory and the expectation theory explain reasonably well the mechanisms of the placebo effect, especially in conjunction with each other. The underlying biochemical pathway, according to the limited current knowledge, involves endorphins for pain and dopamine for Parkinson's disease. Finally, human factors such as the doctor's positive attitudes and a good doctor-patient relationship seem to be more essential than the placebo itself in eliciting the placebo effect. CONCLUSIONS: Given the body of evidence supporting the existence of significant placebo effect and the importance of the doctor-patient relationship in determining it, the human factors of the medical treatment should be emphasised in order to maximise the placebo effect and consequently the overall therapeutic effect of the healing acts. PMID- 16358119 TI - [Psychiatric Residence: findings from a national survey disagree with the National Committee of Medical Residency]. PMID- 16358120 TI - [Expansion of the Family Health Program (FHP) and identification of alcohol related problems in Brazil]. PMID- 16358121 TI - [Schizophrenia and body image distortion: nosological considerations]. PMID- 16358122 TI - ["Quality of life in siblings of autistic patients"--I]. PMID- 16358123 TI - [Topiramate reduces irritability and self-injuries in autistic children]. PMID- 16358125 TI - [Sustained leukopenia probably related to quetiapine]. PMID- 16358126 TI - [Home visit in the treatment of alcohol dependent patients: preliminary data]. PMID- 16358127 TI - [Plastic surgery and eating disorders]. PMID- 16358131 TI - Functional and radiographic evaluation of 214 aggressive benign bone lesions treated with curettage, cauterization, and cementation: 24 years of follow-up. AB - PURPOSE: Treatment with curettage, cauterization, and methylmethacrylate of aggressive benign bone lesions is a method now widely accepted in most orthopedic oncology centers. However, one of the controversies regarding this technique is the possible complications that may arise from the use of methylmethacrylate, which has caused some authors to remove it 2 years after the surgery and replace it with bone graft. The objective of this paper is to present a functional and radiographic evaluation of 214 patients presenting with aggressive benign bone lesions treated with curettage, cauterization, and methylmethacrylate from 1974 to 1998, with some of them having 24 years of follow-up. These patients were clinically and radiographically evaluated for the incidence of late osteoarthrosis, range of motion, and pain in the involved joint. METHOD: This study comprised 214 patients, with an average follow-up duration of 10.6 years (range: 2 to 24 years. All cases involved aggressive benign lesions. The patients were evaluated according to the Musculoskeletal Tumor Society Score (MSTS) functional evaluation system, and the complications are described. RESULTS: The MSTS functional evaluation was excellent in 166 cases (78%), good in 26 (12%), fair in 11 (5%), and poor in 11 (5%). The following complications were observed: late osteoarthrosis, 25 cases (12%); infection, 12 (6%); pathologic fracture, 11 (5%); and local recurrence, 19 (9%). CONCLUSION: Based on clinical assessment, no significant deleterious effects directly related to the use of methylmethacrylate were observed. The functional evaluation performed in 1998 (up to 24 years of follow-up) did not show significant change when compared to the evaluation performed in 1985. PMID- 16358132 TI - Accuracy of magnetic resonance in identifying traumatic intraarticular knee lesions. AB - PURPOSE: To evaluate the diagnostic accuracy of magnetic resonance imaging of the knee in identifying traumatic intraarticular knee lesions. METHOD: 300 patients with a clinical diagnosis of traumatic intraarticular knee lesions underwent prearthoscopic magnetic resonance imaging. The sensitivity, specificity, positive predictive value, negative predictive value, likelihood ratio for a positive test, likelihood ratio for a negative test, and accuracy of magnetic resonance imaging were calculated relative to the findings during arthroscopy in the studied structures of the knee (medial meniscus, lateral meniscus, anterior cruciate ligament, posterior cruciate ligament, and articular cartilage). RESULTS: Magnetic resonance imaging produced the following results regarding detection of lesions: medial meniscus: sensitivity 97.5%, specificity 92.9%, positive predictive value 93.9%, positive negative value 97%, likelihood positive ratio 13.7, likelihood negative ratio 0.02, and accuracy 95.3%; lateral meniscus: sensitivity 91.9%, specificity 93.6%, positive predictive value 92.7%, positive negative value 92.9%, likelihood positive ratio 14.3, likelihood negative ratio 0.08, and accuracy 93.6%; anterior cruciate ligament: sensitivity 99.0%, specificity 95.9%, positive predictive value 91.9%, positive negative value 99.5%, likelihood positive ratio 21.5, likelihood negative ratio 0.01, and accuracy 96.6%; posterior cruciate ligament: sensitivity 100%, specificity 99%, positive predictive value 80.0%, positive negative value 100%, likelihood positive ratio 100, likelihood negative ratio 0.01, and accuracy 99.6%; articular cartilage: sensitivity 76.1%, specificity 94.9%, positive predictive value 94.7%, positive negative value 76.9%, likelihood positive ratio 14.9, likelihood negative ratio 0.25, and accuracy 84.6%. CONCLUSION: Magnetic resonance imaging is a satisfactory diagnostic tool for evaluating meniscal and ligamentous lesions of the knee, but it is unable to clearly identify articular cartilage lesions. PMID- 16358133 TI - External aggression to the limb as a predictive factor in the evolution of patients undergoing arterial revascularization. AB - PURPOSE: A variety of predictive factors for the evolution of arterial grafts in patients with critical ischemia have been well defined in clinical studies, including diabetes mellitus, dialytic renal insufficiency, smoking, and distal arterial runoff. The goal of this study was to determine whether patients with critical ischemia undergoing arterial reconstruction in which ischemic lesions appeared spontaneously, compared to those in which the ischemic lesion appeared following an external aggression to the limb present different patterns of evolution. METHODS: From February 2002 to January 2004, 100 patients undergoing infra-inguinal arterial reconstruction were followed. They were divided into 2 groups: 1) the spontaneous group (n = 52), comprising individuals presenting with ischemic lesions of spontaneous origin and 2) the external aggression to the limb group (n = 48), comprising individuals for which an external causal mechanism for the appearance of the ischemic lesion was identified. The variables analyzed were limb salvage and graft functioning rates. RESULTS: Patients with spontaneous lesions had rates of limb salvage and graft functioning significantly lower than those for patients with lesions that were secondary to external aggression (42.3% versus 87.5%, respectively for both outcomes; P <.001). CONCLUSIONS: The absence of an external aggression as a contributing factor to a critical ischemic lesion in the lower limb may result in a poorer evolution of both graft function and limb salvage following arterial revascularization. However, this factor is not expected to directly influence the case conduct, since almost half of the patients without evident external aggression had good graft functioning and limb salvage. This prognostic factor should be used just as all others are, i.e., to give patients and doctors a better idea of the possible evolution in such cases. PMID- 16358134 TI - Perceptions of patients about cataract. AB - OBJECTIVE: To identify in adult patients suffering from cataract the perceptions regarding the disease and its surgical treatment. MATERIALS AND METHODS: An exploratory survey was conducted among adult patients suffering from cataract and participating in a large-scale cataract management program at the University of Sao Paulo General Hospital in 2004. The interviews were conducted by research assistants previously trained to pose questions and record answers. RESULTS: The sample consisted of 170 men and women (43.5% and 56.5%, respectively), aged between 40 and 88 years. Of the 170 participants, 43.5% were from the State of Sao Paulo, 14.7% from the State of Bahia, 12.4% from the State of Minas Gerais, 5.9% from the State of Pernambuco, 1.8% from other countries, and the remaining 21.7% were from other Brazilian states. Of those who were actively working (n = 87), 43.7% had an occupational level corresponding to nonspecialized manual labor, 27.6% were in specialized manual labor jobs, 25.3% had routine nonmanual occupations, 1.1% supervised manual labor, and 2.3% had low-ranking supervision or inspection jobs over nonmanual occupations. Of those who were not actively working (n = 82), 53.6% were retired, 45.2% were housewives, and 1.2% were unemployed. Concerning conceptions about cataract, 79.0% referred to it as "a small skin fold that gradually covers the eye" and 32.4% mentioned, in addition, other conceptions. Concerning the cause, of the alternatives presented to them, 80% reported aging, 47.1% blamed "overusing the eyes in the workplace or at home", 7.1% believed they had cataract due to some kind of "spell." Of the associated answers, 94.1% referred to "blurred vision" in people suffering from cataract, 72.4% thought the person may become blind, and 66.5% believed that the patients suffering from cataract are depressed because they cannot see. Regarding surgery, 28.8% were afraid of undergoing surgery; of those, 16.3% cited with the fear of dying during surgery, 55.1% thought they might become blind, 40.8% believed the surgery would be painful, and 8.2% followed religious practices that do not permit surgery. CONCLUSION: Some misconceptions were identified, and the fear of blindness was the most mentioned reason for not seeking cataract surgery, which indicates the need for orientation. PMID- 16358135 TI - Frequency of the allelic variant (Trp8Arg/Ile15Thr) of the luteinizing hormone gene in a Brazilian cohort of healthy subjects and in patients with hypogonadotropic hypogonadism. AB - PURPOSE: To evaluate the frequency of allelic variant Trp8Arg/Ile15Thr in the luteinizing hormone beta-subunit gene in a Brazilian population of healthy subjects and in patients with hypogonadotropic hypogonadism. SUBJECTS AND METHODS: Two hundred and two adults (115 women) with normal sexual function and 48 patients (24 women) with hypogonadotropic hypogonadism underwent a molecular study of the the luteinizing hormone beta-subunit gene using a polymerase chain reaction technique followed by enzymatic digestion with the restriction enzymes Nco I (for detection of the Trp8Arg point mutation) and Fok I (for detection of the Ile15Thr point mutation). Basal luteinizing hormone and FSH, testosterone, or estradiol levels were measured in 37 healthy subjects (21 women) and in 27 hypogonadotropic hypogonadism patients (13 women) by immunofluorometric methods (hLH-Spec and hFSH-Spec, AutoDELFIA, Wallac Oy, Turku, Finland). RESULTS: The genetic variant of the luteinizing hormone beta-subunit gene was present at a similar frequency in healthy subjects (14.4%) compared to patients with hypogonadotropic hypogonadism (16.6%). There was no effect of the allelic variant of the luteinizing hormone beta-subunit gene on luteinizing hormone levels in patients with hypogonadotropic hypogonadism as compared to healthy subjects. CONCLUSION: This study indicates that the allelic variant Trp8Arg/Ile15Thr of the luteinizing hormone beta-subunit gene is a common polymorphism in the Brazilian population (14.4%). The same frequency of this luteinizing hormone variant in the groups with hypogonadotropic hypogonadism and in the healthy subjects excludes a relationship between this variant and hypogonadotropic hypogonadism. PMID- 16358136 TI - The influences of positive end expiratory pressure (PEEP) associated with physiotherapy intervention in phase I cardiac rehabilitation. AB - PURPOSE: To evaluate the effects of positive end expiratory pressure and physiotherapy intervention during Phase I of cardiac rehabilitation on the behavior of pulmonary function and inspiratory muscle strength in postoperative cardiac surgery. METHODS: A prospective randomized study, in which 24 patients were divided in 2 groups: a group that performed respiratory exercises with positive airway expiratory pressure associated with physiotherapy intervention (GEP, n = 8) and a group that received only the physiotherapy intervention (GPI, n = 16). Pulmonary function was evaluated by spirometry on the preoperative and on the fifth postoperative days; inspiratory muscle strength was measured by maximal inspiratory pressure on the same days. RESULTS: Spirometric variables were significantly reduced from the preoperative to the fifth postoperative day for the GPI, while the GEP had a significant reduction only for vital capacity (P < .05). When the treatments were compared, smaller values were observed in the GPI for peak flow on the fifth postoperative day. Significant reductions of maximal inspiratory pressure from preoperative to the first postoperative day were found in both groups. However, the reduction in maximal inspiratory pressure from the preoperative to the fifth postoperative day was significant only in the GPI (P < .05). CONCLUSIONS: These data suggest that cardiac surgery produces a reduction in inspiratory muscle strength, pulmonary volume, and flow. The association of positive expiratory pressure with physiotherapy intervention was more efficient in minimizing these changes, in comparison to the physiotherapy intervention alone. However, in both groups, the pulmonary volumes were not completely reestablished by the fifth postoperative day, and it was necessary to continue the treatment after hospital convalescence. PMID- 16358137 TI - Late postoperative follow-up of patients undergoing subtotal splenectomy. AB - PURPOSE: Over the past 21 years, we have performed more than 200 subtotal splenectomies, in which the upper splenic pole vascularized only by the gastrosplenic pole vascularized only by the gastrosplenic vessels is preserved, to treat different pathologic conditions. A meticulous follow-up of the postoperative results of this procedure is of fundamental importance. METHODS: All patients undergoing subtotal splenectomy were invited to be reviewed. A total of 86 patients who had undergone surgery 1 to 20 years ago were gathered; the surgical procedure was performed for one of the following conditions: portal hypertension due to schistosomiasis (n = 43), trauma (n = 31), Gaucher's disease (n = 4), myeloid hepatosplenomegaly due to myelofibrosis (n = 3), splenomegalic retarded growth and sexual development (n = 2), severe pain due to splenic ischemia (n = 2) and pancreatic cystadenoma (n = 1). Patients underwent a hematologic exam, an immunologic assessment, abdominal ultrasonography, computed tomography, scintigraphy and endoscopy. RESULTS: Increased white blood cell count and platelets were the only hematological abnormalities. No immunologic deficit was found. Esophageal varices were still present in patients who underwent surgery because of portal hypertension although without rebleeding. The ultrasound, tomography and scintigraphy exams confirmed the presence of functional splenic remnants without significant size alteration. CONCLUSIONS: Subtotal splenectomy seems to be a safe procedure that can be useful in treating conditions involving the spleen. The functions of the splenic remnants are preserved during long periods of time. PMID- 16358138 TI - Inspiratory muscle training is ineffective in mechanically ventilated critically ill patients. AB - PURPOSE: Invasive mechanical ventilation is associated with complications, and its abbreviation is desirable. The imbalance between increased workload, decreased inspiratory muscle strength and endurance is an important determinant of ventilator dependence. Low endurance may be present due to respiratory muscle atrophy, critical illness, or steroid use. Specific inspiratory muscle training may increase or preserve endurance. The objective of the study was to test the hypothesis that inspiratory muscle training from the beginning of mechanical ventilation would abbreviate the weaning duration and decrease reintubation rate. As a secondary objective, we described the evolution of inspiratory muscle strength with and without inspiratory muscle training. METHODS: Prospective, randomized clinical trial in an adult clinical-surgical intensive care unit. Twelve patients trained the inspiratory muscles twice a day, and 13 patients did not (control). Training was performed adjusting the sensitivity of the ventilator based on the maximal inspiratory pressure. Patients underwent daily surveillance of the maximal inspiratory pressure. RESULTS: The weaning duration (31 +/- 22 hr, control and 23 +/- 11 hr, training group; P = .24) and reintubation rate (5 control and 3 training group; P = .39) were not statistically different. The maximal inspiratory pressure of the control group showed a trend toward a modest increase. In contrast, the training group showed a small decrease (P = .34). CONCLUSIONS: In acute critically ill patients, inspiratory muscle training from the beginning of mechanical ventilation neither abbreviated the weaning duration, nor decreased the reintubation rate. Inspiratory muscle strength tended to stay constant, along the mechanical ventilation, with or without this specific inspiratory muscle training. PMID- 16358139 TI - A furazolidone-based quadruple therapy for Helicobacter pylori retreatment in patients with peptic ulcer disease. AB - PURPOSE: Many of the currently used eradication regimens against Helicobacter pylori fail to cure the infection either due to antimicrobial resistance or to poor patient compliance. The infection leads to increased risk of developing potentially severe complications, such as gastric cancer. This study was aimed at assessing the efficacy and safety of a quadruple therapy with furazolidone for H. pylori retreatment. METHODS: Patients who had failed one or more eradication regimens against H. pylori infection underwent upper gastrointestinal endoscopy. Biopsy specimens were taken from the gastric antrum and corpus for histology and for a urease test and. Patients received omeprazole 20 mg, bismuth citrate 240 mg, tetracycline 500 mg, and furazolidone 200 mg, all twice daily for 7 days. Therapeutic success was evaluated by endoscopy and biopsies 8 weeks after the end of treatment. RESULTS: Sixty two patients were enrolled, and 58 completed the study. Under this protocol, H. pylori eradication was achieved in 39/58 patients (67%). Mild adverse events were reported. CONCLUSION: The short quadruple therapy course with furazolidone is well tolerated, inexpensive, and effective in retreatment for H. pylori infection. It is a good option for developing countries. PMID- 16358141 TI - Emergency contraception and ectopic pregnancy: report of 2 cases. PMID- 16358140 TI - Blunt cerebrovascular injuries. AB - Over the past decade, the recognition and subsequent management of blunt cerebrovascular injuries has undergone a marked evolution. Originally thought to be a rare occurrence, blunt cerebrovascular injuries are now diagnosed in approximately 1% of blunt trauma patients. The recognition of a clinically silent period allows for angiographic screening for injuries based upon the mechanism of trauma and the patient's constellation of injuries. Comprehensive screening of patients has resulted in the early diagnosis of blunt cerebrovascular injuries during the asymptomatic phase, thus allowing treatment that could prevent neurologic sequelae. Although the ideal regimen of antithrombotic therapy is yet to be determined, treatment with either antiplatelet or anticoagulant agents has been shown to reduce the blunt cerebrovascular injuries related stroke rate. Blunt cerebrovascular injury is a rare but potentially devastating injury; appropriate angiographic screening in high-risk patients should be performed and prompt treatment initiated to prevent ischemic neurologic events. PMID- 16358142 TI - Sigmoid colon adenocarcinoma: 9 years from diagnosis to treatment. PMID- 16358143 TI - A new method for scoring active sweat glands. PMID- 16358144 TI - [Health indicators in Brazil]. PMID- 16358145 TI - [Balanced cardiovascular disease mortality from 1980 to 1999--Brazil]. AB - OBJECTIVE: To compare trends in mortality rates from cardiovascular diseases (CVD), ischemic heart diseases (IHD) and cerebrovascular diseases (CBVD) in the States of Rio de Janeiro (RJ), Sao Paulo (SP) and Rio Grande do Sul (RS) and respective capitals from 1980 to 1999. METHODS: Data regarding CVD deaths were obtained from Datasus, and those regarding populations were obtained from IBGE. Crude and sex and age-adjusted mortality rates were calculated using the direct method (standard population: State of Rio de Janeiro's population twenty years of age or older in 2000). Because of the relevant increase in mortality from ill defined causes in the city and State of RJ as of 1990, the deaths were balanced prior to adjustments. The trends were analyzed using linear regressions. RESULTS: Annual declines of balanced and adjusted mortality ranged from -11.3 CVD deaths/100,000 inhabitants in the city and State of RJ to -7.4 in the city of SP. IHD mortality rates were similar in the State and city of RJ and in Porto Alegre, and lower in the city of SP (-2.5 deaths/100,000 inhabitants). CBVD mortality rates ranged from -6.0 to -2.8 deaths/100,000 inhabitants in the State of RJ and in Porto Alegre, respectively. CONCLUSION: A decline in balanced and adjusted CVD, IHD and CBVD mortality rates was observed from 1980 to 1999 in the three States and capitals. In the State and city of RJ declines in IHD were clear as of 1990, whereas declines in CBVD occurred throughout the period studied. PMID- 16358146 TI - [Serial clinical and echocardiographic evaluation in children with Marfan syndrome]. AB - OBJECTIVE: To describe the clinical cardiac manifestations and temporal evolution of Marfan syndrome in children; to estimate the incidence of annuloaortic ectasia and mitral valve prolapse; and to evaluate tolerability and efficacy of beta blockers in these patients. METHODS: During one year, 21 children with Marfan syndrome underwent serial clinical and echocardiographic examinations. Echocardiograms assessed: the presence of mitral valve prolapse, aortic root diameter, mitral and aortic valves regurgitation, and aortic enlargement during beta-blocker therapy. Eleven patients had two measurements of the aortic root taken one year apart. RESULTS: The children were asymptomatic throughout the study. Mitral prolapse was found in 11 (52%) children. Annuloaortic ectasia occurred in 16 (76%) patients and found to be mild in 42.8%, moderate in 9.5%, and severe in 23.8%. One of these patients underwent aortic valve replacement and repair of the ascending aorta by the Bentall-De Bono technique, with good results. Heart rate decreased by 13.6% (from 85 to 73 bpm; p < 0.009) with the use of beta-blockers; however, aortic root diameter increased by 1.4 mm/year (p < 0.02). One child could not be given beta-blockers due to bronchial asthma, and no significant side effects were observed in the remaining children, including one who also had bronchial asthma. CONCLUSION: The children remained asymptomatic throughout the study, the use of beta-blockers led to a significant decrease in heart rate, and no significant adverse effects were observed. Contrary to the literature, incidence of annuloaortic ectasia was high among the study population, greater than that of mitral valve prolapse, even during beta-blocker therapy. PMID- 16358147 TI - [Depression and Acute Coronary Syndromes: gender-related differences]. AB - OBJECTIVE: Investigate whether social and demographic characteristics, clinical diagnosis, smoking status, alcohol consumption, and anxiety are associated with depression in patients with Acute Coronary Syndromes (ACS) stratified by gender. METHODS: Three hundred forty-five consecutive patients with unstable myocardial ischemic syndrome (206 with myocardial infarction and 139 with unstable angina) were interviewed. The interviews included questions about sociodemographics, smoking status, screening for depression (Prime MD e BDI), trait and state anxiety, (IDATE), and alcohol consumption (AUDIT). RESULTS: Diagnosis of depression has significantly correlated with female gender, age under 50 years, and higher average scores on trait anxiety and state anxiety. Depressed men (245) were usually younger than 50 years of age, smokers and had higher average score on trait anxiety and state anxiety than those non-depressed. The multivariate analysis highlights that age is negatively associated with depression (OR 0.9519 95% CI 0.9261 - 0.9784) and that higher scores on trait anxiety are positively associated (OR 1.0691 95% CI 1.0375 - 1.1017) with depression in the male gender. In the female sample (100), depressed women differ from non-depressed women in that they have a higher average score on trait anxiety and state anxiety. In the multivariate analysis of the female sample, a higher score on trait anxiety was independently associated with depression (OR 1.1267 95% CI 1.0632 - 1.1940). CONCLUSION: It was concluded that, among hospitalized patients with ACS, women, as well as men under 50 years and who suffer from anxiety are more likely to experience depression. PMID- 16358148 TI - [Heart failure mortality trend in Salvador, Bahia, Brazil]. AB - OBJECTIVE: To assess mortality trend due to heart failure (HF) in Salvador- Bahia, from 1979 to 1995. METHODS: HF was defined by notations from the 9th Review of International Disease Code (IDC9) 428.0, 428.1 and 428.9. HF death and population data (metropolitan area of Salvador) were obtained by means of Secretaria de Saude da Bahia (Bahia State Health Secretariat) and Instituto Brasileiro de Geografia e Estatistica (Brazilian Institute of Geography and Statistics). Mortality rates (/100,000) were total or per gender and age, and gross or adapted per age (straight standardization). RESULTS: Mortality rates due to HF had a progressive reduction in the period of time assessed, for both genders, especially up to 1992. From then and up to 1995, there was an apparent stabilization of the curves. Gross mortality rate went from 25.0/10(5), in 1979, to 16.4/10(5) inhabitants, in 1995 (a decrease of 34.4%). The reduction was 34.0% (23.3/10(5), in 1979, to 15.4/10(5) inhabitants, in 1995) for male sex and 35.2% (26.7/10(5), in 1979, to 17.3/10(5) inhabitants, in 1995), for female sex. The same trend took place in several age ranges, including the population > or = 40 years old, which has a greater risk for HF. After adaptation per age (standard population of 1979), it is observed that relative reductions in the rates were even greater. CONCLUSION: Mortality due to HF, in Salvador-Bahia, decreased from 1979 to 1992, becoming stable from then to 1995. PMID- 16358149 TI - [Mechanism of cardiac resynchronization therapy by real-time three-dimensional echocardiography in patients with heart failure]. AB - We report the case of a 66-year-old man with heart failure NYHA class IV treated with biventricular pacing for cardiac resynchronization. The patient was evaluated by real-time three-dimensional transthoracic echocardiography before and 48 hours after pacemaker implantation. The use of three-dimensional echocardiography contributed to understanding the underlying mechanism involved in cardiac resynchronization therapy by demonstrating enhanced synchrony of myocardial segments, which resulted in the patient's clinical improvement. PMID- 16358150 TI - [Left internal thoracic artery to left pulmonary artery fistula after coronary artery bypass graft surgery. A rare cause of myocardial ischemia]. AB - We report a patient who developed dyspnea on mild exertion six years after coronary artery bypass graft surgery (CABG). Myocardial ischemia was documented by radionuclide imaging, and coronary angiography showed patency of all grafts and a large fistula between the left internal thoracic artery (LITA) and the left pulmonary artery (LPA). The patient was submitted to surgical closure of the fistula and made an excellent recovery. PMID- 16358151 TI - [Coronary artery aneurysm one year and five months after sirolimus-eluting stent placement]. AB - A 52-year-old man with diagnosis of post-infarction unstable angina. Coronary angiography revealed 90% luminal obstruction in the middle third of the right coronary artery and 90% in the marginal branch of the circumflex artery. After the administration of clopidogrel 300 mg associated with acetylsalicylic acid, the patient underwent the implantation of a sirolimus-eluting stent (CYPHER; Johnson & Johnson--Cordis) 2.5 x 1.8 mm in the lesion located at the left marginal branch. One year and five months after the CYPHER stent implantation, a new angiography showed intrastent coronary aneurysm at the left marginal branch. This case report suggests that the implantation of a sirolimus-coated coronary stent may cause late formation of coronary artery aneurysm. PMID- 16358152 TI - [Mycotic femoral pseudoaneurysm after 11 years of catheterism]. AB - The authors report the unusual case of a 52-year-old mountain climber that presented with pain and swelling in his right thigh which revealed to be a ruptured mycotic pseudoaneurysm with no history of recent trauma or other apparent cause. The patient reported a past history of myocardial infarction 11 years before, with the performing of two femoral catheterisms for coronary angiography. He denied any episode of fever or diagnosis of bacteraemia at that time or later, nor any other complaint during these 11 years. The rarity of the case, the appearance of this extremely late complication together with the patient's kind of sportive activity prompted us to publish the case. PMID- 16358153 TI - [Aorta-right ventricle fistula. An unexpected complication of bacterial endocarditis]. PMID- 16358154 TI - [Treating hypertension in the doctor's office]. PMID- 16358155 TI - [Visiting immunology]. PMID- 16358156 TI - [Cardiovascular manifestations in patients infected with the human immunodeficiency virus]. PMID- 16358159 TI - Pulmonary artery sarcoma misdiagnosed as chronic thromboembolic pulmonary hypertension. AB - Pulmonary artery sarcomas are rare neoplasms of the pulmonary artery that are often confused with chronic thromboembolic disease, as both diseases have similar presentations. In patients with presumed chronic thromboembolic pulmonary hypertension, certain clinical and imaging characteristics may suggest the alternative diagnosis of pulmonary artery sarcoma. In this article we present a case of a man initially diagnosed with chronic thromboembolic pulmonary hypertension, but who was later found to have pulmonary artery sarcoma. We review the distinguishing characteristics of the two diseases and discuss possible treatment strategies. PMID- 16358160 TI - Elevated homocysteine levels in patients with end-stage renal disease. AB - PURPOSE: To examine the effect of hemodialysis on plasma homocysteine levels, and the relationship of these values to clinical cardiovascular events in patients with end-stage renal disease (ESRD). METHODS: Adults undergoing chronic hemodialysis were studied at baseline and at six months. Their clinical histories were obtained at the baseline visit, and measurements of plasma homocysteine concentration were made immediately prior to and following routine dialysis. The occurrence of clinical cardiovascular events was assessed over six months. RESULTS: We enrolled 147 patients (85 men and 62 women, age 58 +/- 15 years) who required hemodialysis for 3.4 +/- 3.4 years (mean +/- SD). The median homocysteine level for this population (including both pre- and post-dialysis values) was 17.3 micromoles/L. Mean pre-dialysis plasma homocysteine levels of patients with clinical cardiovascular disease did not differ significantly from those without the disease (22.5 +/- 9.9 vs. 25.4 +/- 24.5 micromoles/L, respectively), nor did post-dialysis levels differ between these populations. During six months follow-up, rates of ischemic events were not related to hyperhomocysteinemia. The difference between mean pre- and post-dialysis homocysteine levels (26.3 +/- 19.7 and 15.6 +/- 11.4 micromoles/L, respectively) and the decline in homocysteine over the course of a single dialysis treatment session (10.3 +/- 10.2 micromoles/L) were highly significant (p<0.0005). CONCLUSIONS: Plasma homocysteine levels were elevated in 82% of 147 patients with ESRD and fell to the normal range in a majority of patients during a single dialysis treatment session. Mean pre-dialysis levels did not change significantly over six months, however, and plasma homocysteine levels did not predict cardiovascular events in this population. There was also a trend towards worse outcomes in patients with lower homocysteine levels, which correlates to findings from recent studies. Further studies are needed to clarify the association between hyperhomocysteinemia and coronary risk in patients with ESRD. PMID- 16358161 TI - Mean frequency of premature ventricular complexes as predictor of malignant ventricular arrhythmias. AB - AIM: The aim was to test the hypothesis that mean frequency of premature ventricular complexes (PVCs) correlates with vulnerability to malignant arrhythmias such as ventricular tachycardia and/or ventricular fibrillation (VT/VF). METHODS: Patients with an implantable cardioverter defibrillator (ICD) device for underlying ischemic or non-ischemic cardiac pathology were selected from a database. Availability of total count of single (s) PVCs and runs (r) of PVCs was the only inclusion criterion. Forty-four subjects (6 females and 38 males) aged 18-74 years (mean 547.1 years), were eligible. All had a European Pacemaker Identification Card (EPIC) documenting left ventricular ejection fraction (LVEF). The frequency of recorded episodes of VT and VF was obtained from ICD memory. RESULTS: Among patients with ischemic heart disease (IHD) and those with IHD and an LVEF of less than 30%, the mean frequency of PVCs was significantly higher in those with subsequent episodes of VT/VF compared to those without subsequent episodes (p < 0.05 for sPVCs and rPVCs in both groups). CONCLUSION: Among patients with IHD, mean frequency of PVCs is a useful marker of vulnerability to potentially fatal arrhythmias and may be a useful tool for the risk stratification of patients. PMID- 16358162 TI - Successful treatment of cervical spinal epidural abscess by combined hyperbaric oxygenation. AB - A 49-year-old man underwent hyperbaric oxygen (HBO) therapy for the treatment of primary spinal epidural abscess. Although the epidural abscess was initially treated with antibiotic (cefozopran) for 5 days, he subsequently developed motor weakness, paresthesia and urinary retention. MRI demonstrated spinal cord compression at the C1-C4 level. HBO therapy was added to the antibiotic regimen, and three days later we found clinical evidence of a response to treatment. Neurological symptoms were relieved 13 days after HBO therapy. This case suggests that HBO therapy is an effective therapeutic adjunct for the treatment of spinal epidural abscess. PMID- 16358163 TI - The role of patient-controlled apparatus for sedation in the emergency department. AB - AIM: Hand trauma is a fairly common cause of emergency unit admissions. Various analgesic and sedative agents are used to decrease pain and anxiety during minor surgical procedures for hand trauma patients and provide more comfortable conditions for the surgeon. The aim of this study was to investigate the potential role of patient-controlled sedation (PCS) during surgical procedures done under local anesthesia for hand trauma in the emergency department. MATERIALS AND METHODS: Forty ASA I-II (Assignment of the American Society of Anesthesiologists) patients who visited the emergency unit with hand trauma were randomized to 2 groups of 20 patients each. The control group received 1 microg/kg of fentanyl (i.v.) and 0.028 mg/kg of midazolam (i.v.). Additional 1 mg doses of midazolam were given by the anesthesiologist to keep the sedation level between 3 and 4. In the PCS group, the midazolam was administered after programming the apparatus. The settings were as follows: loading dose: 0.028 mg/kg, bolus dose 1 mg, lock-out period: 5 min and basal infusion rate: 0. The loading dose was given before local anesthesia. All patients received prilocaine hydrochloride (Citanest 2%, 10 mL) for local anesthesia. The systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), oxygen saturation (SpO2) and respiration rate (RR) were measured before intervention and at 2, 3, 5, 10, 15, 20 and 30 minutes. RESULTS: There were no differences in the demographic characteristics, operation and discharge times in the two groups (p>0.05). No cardiovascular or respiratory instability was observed in any patient, and SpO2 remained over 95% for all. The SBP, DBP, HR and SpO2 did not differ significantly (p>0.05). Although the sedation levels of all patients were satisfactory, the sedation levels of the control group were significantly lower at 5 and 15 minutes (p<0.05). The total midazolam dose was 4.3+1.1 in the control group and 4.0+0.8 in the PCS group. The patient satisfaction rate was 95% in the PCS group and 80% in the control group (p>0.05). CONCLUSIONS: The two regimens did not differ with respect to hemodynamic changes, sedation levels and patient satisfaction. Therefore, PCS may be an acceptable alternative for surgical procedures performed using local anesthesia. PMID- 16358164 TI - Carbofuran poisoning among farm workers. AB - Accidental or intentional ingestion of carbofuran can produce a life-threatening syndrome that requires prompt diagnosis and treatment. This paper investigates unintentional carbofuran poisoning in farm workers. Thirteen patients were admitted to the emergency department with carbofuran poisoning between January 2002 and August 2004 (2 female, 11 male). The patients had been poisoned while mixing the liquid form of carbofuran with seeds. Their hands were red on admission. Complaints most commonly reported by the patients on admission were nausea, vomiting, headache, weakness, dizziness and blurred vision. The most commonly observed signs were tachycardia, tachypnea, salivation, miosis, elevated blood pressure, and fasciculation. Three patients were agitated and one was lethargic on admission. We reviewed the patients' medical charts retrospectively, as well as the demographic data, intoxication route, clinical and laboratory presentations, and outcomes. We made the diagnosis according to a compatible exposure history and clinical findings. The most commonly observed laboratory finding was hyperglycemia, which was found in 6 patients. Serum pseudocholinesterase level was low in only one patient. All the patients were cured and discharged from the hospital in good physical condition. Rapid onset, mild illness and quick recovery are typical characteristics of acute occupational carbofuran poisoning. We conclude that public health efforts should educate farm workers about the dangers of pesticide application so that its threat can be diminished. PMID- 16358165 TI - Frequency and mortality risk factors of acute adult poisoning in Adana, Turkey, 1997-2002. AB - OBJECTIVE: The purpose of this study was to evaluate the data on acute adult poisoning over a six-year period at a university emergency department in Turkey and identify the risk factors of mortality. MATERIAL AND METHODS: In this survey, data were collected from 2,229 adult patients admitted to the Emergency Department (ED) of Cukurova Medical Faculty, Adana, from January 1, 1997 to December 31, 2002. RESULTS: Of all emergency admissions during the six years, 1.6% were poisoned. Of the total poisoning cases, 725 (32.5%) were males and 1,504 were females. Mean age was 29.3 +/- 13.2 for males and 23.8 +/- 9.6 for females (p=0.001). A majority of the cases (76.4%) were attempted suicides. The attempted suicide ratio is higher for women (gender ratio: 2.4/1) and youths. Drugs were the most frequent cause of poisoning (59.0%) and pesticides were the second (26.4%). Psychoactive drugs were the most common agent (33.5%) among drugs, with their rate increasing each year. Eighty-seven patients died (3.9%): alcohol (methanol) was the most frequent cause of death (20.6%), followed by mushrooms (11.5%), carbon monoxide (10.3%) and pesticides (8.3%). More men than women died by poisoning (OR: 1.8 CI: 1.1-3.1). Gender, age, season, clinical status, initial emergency care and type of substance were significant risk factors for mortality. CONCLUSION: Poisoning by psychoactive drugs was increasing each year of the study, but many drugs are not a serious risk factor for mortality. However, organic phosphorus is a serious risk factor for poisoning and mortality in this area. Providing satisfactory public and emergency staff assistance and education on the subject may reduce the mortality rate. PMID- 16358166 TI - Spontaneous cholecystocutaneous fistula presenting with an abscess containing multiple gallstones: a case report. AB - Spontaneous cholecystocutaneous fistula is a rare complication of chronic calculous cholecystitis. This complication, a result of the natural history of gallbladder stones, was formerly common. Today it is rare, because of early diagnosis and treatment of biliary tract diseases. We report a case of spontaneous cholecystocutaneous fistula in a 70-year-old female patient who presented with an abscess formation in the right upper quadrant. After the incision of this infective focus, many gallstones were picked up. One-stage open cholecystectomy and excision of the fistula tract were carried out after control of the abdominal wall infection. PMID- 16358167 TI - Intestinal obstruction due to rectal endometriosis. AB - We report a case of a premenopausal woman with severe constipation causing intermittent obstruction. Colonoscopy revealed a tight rectal stricture; however, mucosal biopsies were normal. Exploratory surgery revealed an intense fibrotic reaction involving the rectum and uterus, necessitating a simultaneous low anterior resection and hysterectomy. Pathology established a diagnosis of endometriosis. Preoperative diagnosis of rectal endometriosis can be difficult to establish. Endometrial deposits do not invade the mucosa; therefore, colonoscopy with biopsies are frequently non-diagnostic. Surgery may be the only definitive way to obtain a certain diagnosis. In cases involving rectal strictures of unknown etiology in premenopausal women, rectal endometriosis must be included in the differential diagnosis. PMID- 16358168 TI - Acute myocardial infarction after hydrochloric acid ingestion. AB - We report a case of acute ST segment elevated myocardial infarction associated with hydrochloric acid ingestion. Severe systemic acidosis developed shortly after massive hydrochloric ingestion; it was complicated by the presence of acute myocardial infarction. A new complication of acid ingestion is presented and a possible mechanism is discussed. PMID- 16358170 TI - The neurological implications of fibromuscular dysplasia. AB - Fibromuscular dysplasia is a noninflammatory segmental disease of the arteries, of unknown origin. While the renal arteries are most commonly affected, other larger vessels, including the carotid and vertebrals, may be involved. Fibromuscular dysplasia has been implicated as a cause of stroke in adults and (on rare occasions) in children. The classic angiographic presentation is that of a string of beads. The condition frequently requires surgical or interventional radiological treatment. These approaches most commonly lead to long-term neurological improvement. Five cases with typical angiographic appearance are presented. PMID- 16358171 TI - Internal carotid artery dissection with associated oculosympathetic palsy. PMID- 16358173 TI - [You are a physician in Germany]. PMID- 16358174 TI - [The patient as partner -- observations regarding a respectful relationship with sick people]. PMID- 16358175 TI - [Odor and smelling -- or: what is the meaning of an olf?]. PMID- 16358176 TI - [Smoking -- the futile fight against the vice. A historical perspective]. PMID- 16358177 TI - [Health and anti-aging. Risks and side effects of a new religion]. PMID- 16358178 TI - [From nourishment for the sick to "functional food." Paths and aberrations of mass-produced food from the 19th up into the 21st century]. PMID- 16358179 TI - [God in the mirror? The visibility of the invisible in ancient perception]. PMID- 16358180 TI - [Philosophy and medicine. Do they still have something to say to each other?]. PMID- 16358181 TI - [Poetry and pathology. Literary figures as models for disease terms]. PMID- 16358183 TI - [First results on radial forearm free flap monitoring using indocyaninegreen fluorescence angiography]. PMID- 16358184 TI - [Obituary prof. Dr. Hans-Georg Boenninghaus 20. 4. 1921-7. 5. 2005]. PMID- 16358185 TI - [Editorial comment on the german consensus paper on neonatal hearing screening]. PMID- 16358186 TI - [Universal hearing screening in newborns -- recommendations for organizing and conducting universal hearing screening for congenital hearing loss in Germany]. AB - The Interdisciplinary Consensus Conference for Newborn Hearing Screening (IKKNHS) has worked out joint recommendations for universal hearing screening of newborns. In the consensus paper, 11 professional associations and scientific societies in the fields of gynecology and obstetrics, ENT, pediatrics, and phoniatrics and pedaudiology came to an agreement how to implement newborn hearing screening in Germany. The paper deals with the following topics: goals of universal newborn hearing screening, target group of hearing screening, schedule for screening, personnel involved in the screening program, technologies and framework conditions of hearing screening, documentation, continuous quality control of screening, confirmation diagnostics for conspicuous test subjects, motivation to take part in screening, information on newborn hearing screening, tracking, various infrastructural situations in urban and rural regions, follow-up care, in patient vs. out-patient screening, cost factors of screening, reporting children with permanent hearing loss to the German Central Registry for hearing loss in children. PMID- 16358187 TI - [Guided surgery in a subtemporal approach in CPA tumors?]. AB - BACKGROUND: Computer assisted surgery (CAS) permits the visualization of hidden bony covered structures invisible for the human eye with radiological 3d data sets. The surgeon might be able to orientate anatomically during surgery without having to prepare the according landmarks. This would mean less surgical traumatization and a shorter and smaller operation corridor. METHOD: We determined the use of CAS in a quality assurance analysis with the subtemporal approach in 8 patients with supra-meatal tumors type A of the cerebellopontine angle. Various navigation systems and methods for referencing for the registration of the patients' heads were used. The question was whether it is possible intraoperatively without preparation of known anatomical landmarks to define the borders of an optimal positioned temporary bone cap and to identify the bony covered inner auditory canal and its neuronal structures without orienting neurostimulation. RESULT: It was possible with CAS to assess intraoperatively the borders of a temporary bone cap above the cranially positioned mastoid cell. However, the objective inaccuracy of 2 to 28 mm observed during surgery did not allow a secure identification of the inner auditory canal. CONCLUSIONS: CAS with the subtemporal approach cannot replace the conventional preparation of known anatomical landmarks nor neurostimulation to identify neural structures, due to the expected high inaccuracy with the non-invasive referencing systems that are available today. PMID- 16358188 TI - [Orbital complications of acute pediatric rhinosinusitis: medical treatment versus surgery and analysis of the computer tomogram]. AB - BACKGROUND: One of the most common complications of acute pediatric rhinosinusitis is orbital complication. For subperiosteal abscess, in particular, there are no generally accepted therapy recommendations. PATIENTS: A retrospective study of 45 children with acute orbital rhinosinusitis complications, treated on an in-patient basis at the Department of ENT, Inselspital, University of Bern (1999-2004) is presented. The aim was to analyze conservative medical versus surgical treatment with regard to the individual stages of complications (according to the classification of Moloney, 1987) and, subsequently, to arrive at a therapy recommendation. RESULTS: All children with preseptal cellulitis (stage I), 80 % of children with radiological a subperiosteal phlegmona (Stage IIA) and 67 % of the children with radiological a subperiosteal abscess (Stage IIB) could be treated conservatively. The child with the orbital cellulitis (Stage III) was treated surgically. CONCLUSION: The preseptal cellulitis can be treated conservatively. We recommend an initial conservative therapy of 24 - 48 hours for subperiosteal phlegmona and subperiosteal abscess in the presence of normal vision. Surgical intervention should take place only if there is no improvement after this time interval. The orbital cellulitis should be treated primary surgically. PMID- 16358189 TI - [T-cells activated with a trifunctional bi-specific antibody in head and neck cancer]. AB - BACKGROUND: The trifunctional bi-specific antibody Removab bridges and activates CD3 positive T cells to EpCAM on carcinoma cells and simultaneously binds to an accessory immune cell, thus inducing tumor cell lysis. Following intravenous application, Removab may induce cytokine-related side effects resulting in a sepsis like syndrom. It was questioned, if peripheral mononuclear blood cells (PBMN) already opsonized with Removab could retain antitumor activity and induce less cytokine release than the mere antibody. METHODS: PBMN of patients with head and neck cancer were incubated with Removab and the released cytokines were washed out. Then the Removab-opsonized PBMN were coincubated with genuine tumor cells of the same patient on a chorioallantois membrane for 24 h (T 24) and 48 h (T 48). Tumor cells coincubated with Cisplatin or solely cell culture medium served as control. RESULTS: Coincubation of tumor cells with opsonized PBMN resulted in a 32 % decrease of viable cells at T 24 and a 37 % decrease at T 48, whereas viable cells increased by 10 % at T 24 or 3 % at T 48 when incubated with medium alone (p < 0.05). This tumor cytotoxicity was similar to that of Cisplatin (35 % at T 24/37 % at T 48). CONCLUSION: In an autologous human ex vivo tumor system, Removab-opsonized PBMN induce tumor cell lysis with significantly reduced cytokine release after i. v. application. PMID- 16358190 TI - [Rehabilitation of the olfactory sense after laryngectomy -- the larynx bypass]. AB - BACKGROUND: Patients after laryngectomy lose their previous sense of smell, since air normally circulates via the tracheostoma and active air intake through the nose is no longer possible. The larynx bypass with a throat mask is an aid here. Nasal air circulation is made possible by this connecting piece between the mouth and tracheostoma. MATERIALS AND METHODS: Twenty patients (17 men and 3 women, mean age 60 years) who underwent laryngectomy between November 2003 and February 2004 were examined using Sniffin' Sticks with and without the larynx bypass. Moreover, patients were asked about the practicability of the larynx bypass using a visual analog scale (VAS). In addition to larynx bypass function, we were also interested in the handling for the laryngectomized patients. RESULTS: Overall, patients were able to correctly identify more items with the larynx bypass. The results were significantly better with than without the larynx bypass (p < 0.001). However, the first application of the larynx bypass was considered cumbersome. The median VAS was 5. CONCLUSION: Patients had a better sense of smell with the larynx bypass. However, due to its rather moderate practicability, the larynx bypass does not seem to be suited for daily use in its present form. PMID- 16358191 TI - [Finding the appropriate therapy with the help of sleep endoscopy]. AB - Nasal continuous positive airway pressure (nCPAP) ventilation is the gold standard in the treatment of obstructive sleep apnea (OSA). For patients with CPAP-intolerance and for those tolerating the CPAP-mask but without success, surgical strategies are of growing interest. There are several surgical concepts which combine different surgical procedures on both the velopharyngeal and the retro-lingual section of the upper airway. The case presented here describes the failed CPAP-therapy in an OSAS-patient who tolerates the CPAP-mask. The apneas and hypopnoes could not be reduced by using the CPAP-mask, even more they increased under CPAP-ventilation. Performing a video-documented sleep-endoscopy of the upper airway (under physiological sleep patterns) the location of the obstruction could be identified. After performing a hyoid suspension in combination with a radiofrequency therapy of the base of tongue the apneas and hypopneas decreased. Only by using the sleep endoscopy the reason for the failed CPAP-therapy could be identified and therefore the surgical protocol succeed. To what extend the sleep video endoscopy becomes a standardized procedure in sleep labs warrants further investigations. PMID- 16358192 TI - [Vertigo attacks -- current therapy]. PMID- 16358193 TI - [How does a cochlear implant speech processor work?]. AB - Cochlear implants have become a standard treatment modality for sensorineural hearing loss. In this review article, assembly and function of a cochlear implant are described. Cochlear implants replace the normal inner ear by transforming acoustic sound signals into electric stimuli and deliver these to the auditory nerve. Speech processors translate the acoustic signal of the microphone into one that fits electrostimulation of the auditory system. In multiple steps, the signal has to be analyzed and processed to fit the demands of electrical stimulation. The speech processor then sends commands and the energy for stimulation to the implanted parts via a transcutaneous high frequency radio link. The implant refers the information as electrical stimuli to each electrode contact. PMID- 16358197 TI - [Endonasal micro-endoscopic tumor surgery: state of the art]. AB - BACKGROUND: The aim of the present study was to establish the efficacy of endonasal micro-endoscopic surgery for removal of benign and malignant neoplastic lesions of the paranasal sinuses and the anterior skull base. MATERIAL AND METHODS: This retrospective study evaluated 350 patients with tumors of the paranasal sinuses and the anterior skull base (215 benign and 135 malignant tumors) that were treated surgically at the ENT-Department of the Hospital Fulda gAG between 1993 and 2003. Median follow-up was 65 months. RESULTS: Most frequent entities of benign tumors were osteomas, inverted papillomas and juvenile angiofibromas. Adenocarcinomas, squamous cell carcinomas and esthesioneuroblastomas were most frequently treated in the group of malignomas. 54 % of the benign (n = 118) and 41 % of the malignant tumors (n = 54) were resected exclusively via the endonasal micro-endoscopic approach. Within the follow-up period 3 recurrencies were observed, two inverted papillomas (one was operated endonasally) and one juvenile angiofibroma. In the malignoma group 34 patients died because of tumor disease (16 cases due to recurrencies, 18 cases due to metastases formation). The 5 year survival rate was 66.4 % with respect to the disease-specific survival. The Kaplan-Meier analysis revealed statistically significant differences for the pT stage: pT2 and pT3 tumors have had a 5-year disease-specific survival of 92.3 % and 83.8 %, respectively, compared to 61.5 % of the pT4 tumors. Disease-specific survival also showed differences dependent on histology, tumor site and occurrence, but was without proven significancy. In the patient subgroup who suffered from adenocarcinoma, squamous cell carcinoma or esthesioneuroblastoma the 5-year disease-specific survival was 78.4 % of 29 patients after endonasal resection compared to 66.4 % in 51 patients operated via an external approach. CONCLUSION: The advances in endonasal micro-endoscopic surgery also allow a safe and effective removal of benign and malignant anterior skull base tumors with low morbidity. Thereby, indication is dependent on tumor site and size as well as histology. PMID- 16358198 TI - [Paranasal sinus mucoceles: surgical management and long term results]. AB - BACKGROUND: The aim of this study was to determine the best surgical approach in the treatment of paranasal sinus mucoceles according to their localization. MATERIAL AND METHODS: A retrospective analysis was carried out in 255 patients with 290 sinus mucoceles who were treated surgically at the ENT-Department of the Hospital Fulda gAG between 1983 and 2001. This series include 125 frontal sinus, 23 frontoethmoid, 41 ethmoid, 72 maxillary sinus and 26 sphenoid mucoceles. The patients' history, presenting symptoms, radiological findings, and surgical management were reviewed. Of them, 185 patients were followed by endoscopic and CT or MRI control during a period of up to 19 years, median follow-up was 12 years. RESULTS: In 168 out of 255 patients (66 %) the mucoceles were arisen due to previous sinus surgery, in 37 cases (14 %) after traumatic lesions, in 5 patients (2 %) due to chronic sinusitis and in 2 cases (<1 %) according to tumors. In 43 cases (17 %) no causes were found. In 78.8 % the previous operation was performed via an external approach, either according to Jansen/Ritter or Caldwell-Luc, contrary to 1.5 % after endonasal surgery. The median period until mucocele appearance was 15 years for maxillary sinus, 13 years for frontoethmoid, 10 years for ethmoid, and 8 years for frontal and sphenoid sinus celes, respectively. 201 mucoceles (69.3 %) have been operated endonasal micro endoscopically, 18.6 % via the osteoplastic approach, 10 % endoscopically combined with an osteoplastic procedure and only 2 % according to Jansen/Ritter. Thereafter, recurrence of mucoceles was found in 4 patients only ( = 2.2 %; related to the endonasal approach = 1.6 %). CONCLUSION: Frontoethmoidal, ethmoidal, sphenoidal and maxillary sinus mucoceles are excellent indication for exclusively endonasal micro-endoscopic surgery. The osteoplastic approach combined with endonasal surgery is suitable in far lateral located frontal or maxillary sinus mucoceles. PMID- 16358199 TI - [Reconstruction of the frontoorbital frame using split-thickness calvarial bone grafts]. AB - BACKGROUND: There are many autogenous and allogenous grafts as well as alloplastic materials available for the reconstruction of craniofacial bony frame. We review our techniques and describe the advantages of using calvarial bone, especially split-thickness grafts for bone reconstruction in this area. PATIENTS AND METHODS: Between 1996 and 2003 the orbitocranial bony frames of 15 patients were reconstructed using split calvarial bone grafts at the ENT Department of the Hospital Fulda gAG. In 12 patients the anterior frontal sinus wall or the entire Os frontale were affected. In 1 patient each the lateral wall, roof and floor of the orbit had to be reconstructed. The causes of the bone defects were trauma (n = 6), recurrent frontal sinusitis partly with osteomyelitis (n = 4), benigne tumors (n = 2) and malignancies (n = 3). Twelve patients have had multiple previous operations. In 3 patients the bone reconstruction was performed in the same operation as the tumor removal. Within the follow-up period between 2 and 8 years the split calvarial bone grafts remained stable in size and shape. Graft rejection, osteomyelitis or bone resorption did not occur. Furthermore, we have not experienced significant complications in harvesting cranial bone and have not seen major donor site morbidity. CONCLUSIONS: Our results demonstrate that split-thickness calvarial bone is an excellent graft not only for facial and forehead contouring but also for orbital and complex craniofacial reconstruction. PMID- 16358200 TI - [Management of congenital nasal fistulas and cysts]. AB - BACKGROUND: Congenital nasal fistulas and cysts are uncommon craniofacial malformations. Mostly their symptoms appear already during childhood. In general, they are superficially located but they can reach the skull base or extend intracranially. PATIENTS AND METHODS: Between 1979 and 2004 fifteen patients (11 children and 4 adults) with nasal fistulas and cysts were treated surgically at the ENT-Department of the Hospital Fulda gAG. In 7 patients (46.6 %) the skull base was involved. An intracranial intradural extension was found in 1 child and therefore needed a combined ENT and neurosurgical procedure. In the other 6 cases the nasal fistulas or cysts were located extradurally. They were resected by removing the Christa galli and strenghten the dura. An additional septorhinoplasty was carried out in 4 of the 15 patients. Within the follow-up period of up to 25 years no recurrences were observed. CONCLUSIONS: The adequate therapy of nasal fistulas and cysts consists of complete resection. Magnetic resonance tomography (MRI) and computed tomography (CT) are essential to diagnose intracranial extension and to choose the proper surgical approach including a potential interdisciplinary procedure. PMID- 16358201 TI - [Angiography of the carotid artery in centro-lateral skull base fractures?]. AB - BACKGROUND: In latero-basal, central or spheno-fronto-orbital skull base fractures the internal carotid artery is more frequently involved in severe lesions than expected. PATIENTS AND METHODS: Between 1996 and 2003 we examined 684 patients with Glasgow Coma Scales (GCS) between 2 and 15, median 7.2, using computed tomography (CT). In suspicion of a latero-basal, central or spheno fronto-orbital fractur they got an additional high resolution skull base CT. If the bony canal of the internal carotid artery (ICA) was involved the patient underwent digital subtraction angiography (DSA). The ICA lesion was treated either interventional neuroradiologically, by surgery or only conservatively. RESULTS: Of the 684 patients 33 (4.8 %) had fractures of the ICA bony canal and therefore underwent DSA. Among them were 25 men and 8 women (mean age 35.3 years). Lesions of the ICA were seen in 1.9 % of the patients. A traumatic cavernous-carotid fistula was found in 7 patients (1 %) and in 6 patients (0.9 %) a dissection and/or an aneurysm of the ICA was diagnosed. Six of the patients had clinical symptoms. The lesions were treated primarily interventional neuroradiologically (n = 5) as well as surgically in two cases by clipping the aneurysm and closing the sphenoid sinus, respectively. CONCLUSIONS: Vessel lesions of the ICA in skull base fractures and involvement of the bony carotid canal are more frequent than mentioned in current literature. A solid diagnosis can only be achieved by DSA. Early diagnosis and treatment is important for improving the prognosis of these often multiply injured patients. PMID- 16358202 TI - [Functional results, cognitive and effective quality of life disturbances after trans-temporal resection of acoustic neuroma]. AB - BACKGROUND: The aim of the study was to evaluate postoperative hearing, facial nerve function, quality of life (QOL), affective status and neuropsychological performance after trans-temporal removal of acoustic neuromas (AN). PATIENTS AND METHODS: A retrospective analysis was carried out in 89 patients with AN who were operated at the ENT-Department of the Hospital Fulda between 1988 and 2004. Median follow up was 57 months. Of these 41 patients were additionally examined by a psychologist for evaluation of QOL, neuropsychological functions and affective disorders using several questionnaires. RESULTS: The AN were classified as follows: Type A = 53 %, type B = 35 % and type C = 12 %. One year postoperatively facial nerve function was excellent in 93.3 % of the patients (grade I and II). In 53 % of cases hearing could be preserved. The subjective QOL was expressed through depression and social withdrawal associated with deafness. However, facial nerve dysfunction did not lead to QOL effect. Objective QOL (functional level) was reduced because of verbal memory disturbances and symptoms like dizziness. In 47.5 % of the patients affective and/or neuropsychological dysfunctions were diagnosed. Subsequent MRI evaluation showed lesions of the temporal lobe in 40.5 % of the cases. Of these 80 % suffered from cognitive and/or affective disturbances. CONCLUSIONS: The cognitive and affective disturbances after trans-temporal removal of AN could be to a certain degree due to the elevation of the temporal lobe during surgery. Further research, especially pre- and postoperative examination of QOL and psychological state as well as the comparison between different approaches, particularly trans-temporal vs suboccipital have to clear up specific morbidity of the different approaches. PMID- 16358203 TI - [Management of occult malformations at the lateral skull base]. AB - BACKGROUND: Occult malformations of the lateral skull base are rare anomalies, but can cause severe complications such as recurrent meningitis. Therefore, they need to be precisely delineated and sufficient surgical closure is mandatory. PATIENTS AND METHODS: Between 1986 and 2004 twenty patients (10 children and 10 adults) with occult malformations at the lateral skull base were treated surgically at the ENT-Department of the Hospital Fulda gAG. Of these 3 Mondini malformations, 11 defects of the tegmen tympani or the mastoidal roof, 2 dural lesions to the posterior fossa and 4 malformations within the pyramidal apex have been found. Four patients have had multiple anomalies. Routing symptom was in all cases at least one previous meningitis. Radiological diagnostics included high resolution computed tomography (CT) and magnetic resonance imaging (MRI) as well as CT- or MR-cisternography. Depending on type and localisation of the defect the following surgical algorithm was carried out: The trans-mastoidal approach was used in all cases of Mondini-malformation (including obliteration of the ear), in case of lesions to the posterior fossa as well as partly in anomalies at the tegmen tympani and mastoidal roof, respectively. Defects of the pyramidal apex should be explored via the trans-mastoidal way if the lesion is located caudally to the inner auditory canal (IAC), whereas the trans-temporal approach should be used if the lesion is situated ventral to the IAC and dorso-medially to the internal carotid artery (ICA). The trans-temporal approach was also performed in large defects of the tegmen tympani and mastoidal roof as well as in recurrences. CONCLUSIONS: In all cases of recurrent meningitis caused by agents of the upper airway tract the basic principle should be to search for occult skull base malformations radiologically as well as by sodium fluorescein endoscopy as long as the anomaly is detected. PMID- 16358204 TI - [Cat-scratch disease as cause of Lymphadenitis colli]. AB - Cat-scratch Disease as Cause of Lymphadenitis colli. Cat-scratch disease is a frequent cause of lymphadenitis colli. It mainly affects children and adolescents younger than 21 years. Since the clinical picture is not characteristic, a history of contact to cats or kittens is highly valuable for diagnosing the disease. Major aspects of the disease concerning epidemiology, diagnostic procedures, clinical presentation and therapy are discussed. PMID- 16358214 TI - SLC34A3 mutations in patients with hereditary hypophosphatemic rickets with hypercalciuria predict a key role for the sodium-phosphate cotransporter NaPi-IIc in maintaining phosphate homeostasis. AB - Hereditary hypophosphatemic rickets with hypercalciuria (HHRH) is a rare disorder of autosomal recessive inheritance that was first described in a large consanguineous Bedouin kindred. HHRH is characterized by the presence of hypophosphatemia secondary to renal phosphate wasting, radiographic and/or histological evidence of rickets, limb deformities, muscle weakness, and bone pain. HHRH is distinct from other forms of hypophosphatemic rickets in that affected individuals present with hypercalciuria due to increased serum 1,25 dihydroxyvitamin D levels and increased intestinal calcium absorption. We performed a genomewide linkage scan combined with homozygosity mapping, using genomic DNA from a large consanguineous Bedouin kindred that included 10 patients who received the diagnosis of HHRH. The disease mapped to a 1.6-Mbp region on chromosome 9q34, which contains SLC34A3, the gene encoding the renal sodium phosphate cotransporter NaP(i)-IIc. Nucleotide sequence analysis revealed a homozygous single-nucleotide deletion (c.228delC) in this candidate gene in all individuals affected by HHRH. This mutation is predicted to truncate the NaP(i) IIc protein in the first membrane-spanning domain and thus likely results in a complete loss of function of this protein in individuals homozygous for c.228delC. In addition, compound heterozygous missense and deletion mutations were found in three additional unrelated HHRH kindreds, which supports the conclusion that this disease is caused by SLC34A3 mutations affecting both alleles. Individuals of the investigated kindreds who were heterozygous for a SLC34A3 mutation frequently showed hypercalciuria, often in association with mild hypophosphatemia and/or elevations in 1,25-dihydroxyvitamin D levels. We conclude that NaP(i)-IIc has a key role in the regulation of phosphate homeostasis. PMID- 16358215 TI - Hereditary hypophosphatemic rickets with hypercalciuria is caused by mutations in the sodium-phosphate cotransporter gene SLC34A3. AB - Hypophosphatemia due to isolated renal phosphate wasting results from a heterogeneous group of disorders. Hereditary hypophosphatemic rickets with hypercalciuria (HHRH) is an autosomal recessive form that is characterized by reduced renal phosphate reabsorption, hypophosphatemia, and rickets. It can be distinguished from other forms of hypophosphatemia by increased serum levels of 1,25-dihydroxyvitamin D resulting in hypercalciuria. Using SNP array genotyping, we mapped the disease locus in two consanguineous families to the end of the long arm of chromosome 9. The candidate region contained a sodium-phosphate cotransporter gene, SLC34A3, which has been shown to be expressed in proximal tubulus cells. Sequencing of this gene revealed disease-associated mutations in five families, including two frameshift and one splice-site mutation. Loss of function of the SLC34A3 protein presumably results in a primary renal tubular defect and is compatible with the HHRH phenotype. We also show that the phosphaturic factor FGF23 (fibroblast growth factor 23), which is increased in X linked hypophosphatemic rickets and carries activating mutations in autosomal dominant hypophosphatemic rickets, is at normal or low-normal serum levels in the patients with HHRH, further supporting a primary renal defect. Identification of the gene mutated in a further form of hypophosphatemia adds to the understanding of phosphate homeostasis and may help to elucidate the interaction of the proteins involved in this pathway. PMID- 16358216 TI - A novel framework for sib pair linkage analysis. AB - Sib pair linkage analysis of a dichotomous trait is a popular method for narrowing the search for genes that influence complex diseases. Although the pedigree structures are uncomplicated and the underlying genetic principles straightforward, a surprising degree of complexity is involved in implementing a sib pair study and interpreting the results. Ascertainment may be based on affected, discordant, or unaffected sib pairs, as well as on pairs defined by threshold values for quantitative traits, such as extreme discordant sib pairs. To optimize power, various domain restrictions and null hypotheses have been proposed for each of these designs, yielding a wide array of choices for the analyst. To begin, we systematically classify the major sources of discretion in sib pair linkage analysis. Then, we extend the work of Kruglyak and Lander (1995), to bring the various forms into a unified framework and to facilitate a more general approach to the analysis. Finally, we describe a new, freely available computer program, Splat (Sib Pair Linkage Analysis Testing), that can perform any sib pair statistical test currently in use, as well as any user defined test yet to be proposed. Splat uses the expectation maximization algorithm to calculate maximum-likelihood estimates of sharing (subject to user specified conditions) and then plots LOD scores versus chromosomal position. It includes a novel grid-scanning capability that enables simultaneous visualization of multiple test statistics. This can lead to further insight into the genetic basis of the disease process under consideration. In addition, phenotype definitions can be modified without the recalculation of inheritance vectors, thereby providing considerable flexibility for exploratory analysis. The application of Splat will be illustrated with data from studies on the genetics of diabetic nephropathy. PMID- 16358217 TI - A Y-chromosome signature of hegemony in Gaelic Ireland. AB - Seventeen-marker simple tandem repeat genetic analysis of Irish Y chromosomes reveals a previously unnoted modal haplotype that peaks in frequency in the northwestern part of the island. It shows a significant association with surnames purported to have descended from the most important and enduring dynasty of early medieval Ireland, the Ui Neill. This suggests that such phylogenetic predominance is a biological record of past hegemony and supports the veracity of semimythological early genealogies. The fact that about one in five males sampled in northwestern Ireland is likely a patrilineal descendent of a single early medieval ancestor is a powerful illustration of the potential link between prolificacy and power and of how Y-chromosome phylogeography can be influenced by social selection. PMID- 16358218 TI - Diversity and functional consequences of germline and somatic PTPN11 mutations in human disease. AB - Germline mutations in PTPN11, the gene encoding the protein tyrosine phosphatase SHP-2, cause Noonan syndrome (NS) and the clinically related LEOPARD syndrome (LS), whereas somatic mutations in the same gene contribute to leukemogenesis. On the basis of our previously gathered genetic and biochemical data, we proposed a model that splits NS- and leukemia-associated PTPN11 mutations into two major classes of activating lesions with differential perturbing effects on development and hematopoiesis. To test this model, we investigated further the diversity of germline and somatic PTPN11 mutations, delineated the association of those mutations with disease, characterized biochemically a panel of mutant SHP-2 proteins recurring in NS, LS, and leukemia, and performed molecular dynamics simulations to determine the structural effects of selected mutations. Our results document a strict correlation between the identity of the lesion and disease and demonstrate that NS-causative mutations have less potency for promoting SHP-2 gain of function than do leukemia-associated ones. Furthermore, we show that the recurrent LS-causing Y279C and T468M amino acid substitutions engender loss of SHP-2 catalytic activity, identifying a previously unrecognized behavior for this class of missense PTPN11 mutations. PMID- 16358220 TI - Henoch-Schonlein nephritis. AB - Henoch-Schonlein purpura is a clinical syndrome characterized by the association of skin, joint, and gastrointestinal symptoms. Henoch-Schonlein purpura is characterized by wide-spread vasculitis. Although the clinical symptoms of this disease are characteristic, the diagnosis is not always easy to establish because other forms of systemic vasculitis - mainly the microscopic form of periarteritis nodosa - may mimic the disease. In addition, in contrast to systemic lupus erythematosus, there are no biological tests that can identify Henoch-Schonlein purpura with certainty. Immunofluorescence microscopic techniques have made an important contribution to both the diagnosis and the study of the pathogenesis of the disease, particularly since they have demonstrated the presence of IgA deposits in the glomeruli and in the vessel walls. These findings not only confirmed the immunologic nature of the pathologic lesions but also drew attention to the remarkable similarity between Henoch-Schonlein purpura nephritis and IgA nephropathy. PMID- 16358219 TI - The SERPINE2 gene is associated with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a complex human disease likely influenced by multiple genes, cigarette smoking, and gene-by-smoking interactions, but only severe alpha 1-antitrypsin deficiency is a proven genetic risk factor for COPD. Prior linkage analyses in the Boston Early-Onset COPD Study have demonstrated significant linkage to a key intermediate phenotype of COPD on chromosome 2q. We integrated results from murine lung development and human COPD gene-expression microarray studies with human COPD linkage results on chromosome 2q to prioritize candidate-gene selection, thus identifying SERPINE2 as a positional candidate susceptibility gene for COPD. Immunohistochemistry demonstrated expression of serpine2 protein in mouse and human adult lung tissue. In family-based association testing of 127 severe, early-onset COPD pedigrees from the Boston Early-Onset COPD Study, we observed significant association with COPD phenotypes and 18 single-nucleotide polymorphisms (SNPs) in the SERPINE2 gene. Association of five of these SNPs with COPD was replicated in a case control analysis, with cases from the National Emphysema Treatment Trial and controls from the Normative Aging Study. Family-based and case-control haplotype analyses supported similar regions of association within the SERPINE2 gene. When significantly associated SNPs in these haplotypic regions were included as covariates in linkage models, LOD score attenuation was observed most markedly in a smokers-only linkage model (LOD 4.41, attenuated to 1.74). After the integration of murine and human microarray data to inform candidate-gene selection, we observed significant family-based association and independent replication of association in a case-control study, suggesting that SERPINE2 is a COPD-susceptibility gene and is likely influenced by gene-by-smoking interaction. PMID- 16358221 TI - Role of complement in innate and autoimmunity. AB - The complement system is a major driver of our innate immune response and plays an important role in defence against foreign pathogens. It is composed of a large number of proteins that are found in the circulation, in tissues and other body fluids in pro-enzymatic form. The many biologic activities of the complement system are uncovered following activation by three known pathways. Together with phagocytic cells the complement system is a powerful tool to defend the host against foreign attack. However the complement system, under certain conditions, can be unfavorable to the host namely in diseases such as IgA nephropathy, systemic lupus erythematosus and in diseases where minor changes have occurred in the integrity of tissue such as in ischemia reperfusion. This review describes the many faces of complement in the field of innate and acquired immunity and takes the reader on a journey of the role of complement in health and disease. PMID- 16358222 TI - The klotho gene: a gene predominantly expressed in the kidney is a fundamental regulator of aging and calcium/phosphorus metabolism. AB - A unique short lifespan mouse strain was developed in which a single gene mutation caused multiple aging-related disorders. The gene responsible has been identified as the klotho gene. The most characteristic phenotypes of klotho mice seem caused by abnormalities in calcium metabolism. Klotho plays a critical role in the regulation of calcium and phosphorus homeostasis by negatively regulating active vitamin D synthesis. A defect in klotho gene expression causes the independent impairment both of osteoblast and osteoclast differentiation, leading to low-turnover osteopenia. In humans, two klotho gene polymorphisms were significantly associated with bone density in aged postmenopausal women, but not in pre-menopausal or younger postmenopausal women in two genetically distinct racial populations. The klotho gene is predominantly expressed in the kidney and the expression level of klotho RNA was shown to be greatly reduced in the kidneys of chronic renal failure (CRF) patients. Angiotensin II induced renal klotho down regulation through angiotensin type 1 receptor-dependent but pressor-independent events, while dietary P(i) restriction induced klotho expression. The down regulation of the renal klotho gene could increase renal damage induced by angiotensin II, while klotho gene induction could have therapeutic possibilities in treating angiotensin II-induced kidney damage. PMID- 16358223 TI - The ultrastructural basis of renal pathology in monoclonal gammopathies. AB - The kidney is frequently involved in the course of monoclonal gammopathies (MG). Renal involvement presents different clinical-morphological patterns, which can occur either at the onset or in a late phase of the hematological disease, as well as after chemotherapy. The reasons for the organ tropism of monoclonal immunoglobulins (Igs) are still unknown. Currently, it is well known that some primary structure alterations in monoclonal Igs and/or in their segments correlate to nephrotoxicity. On the other hand, it is impossible to predict the pathogenicity and the clinical manifestations induced by a specific monoclonal Ig based on its specific conformational modifications. Pathogenicity and organ tropism are probably complex phenomena, which involve specific protein factors, patient factors, target organ characteristics and monoclonal plasmacellular mass entities. However, aminoacidic sequence analysis of nephrotoxic Igs and some recent in vitro studies have allowed two different monoclonal light chain (LC) types to be distinguished. Glomerulopathic LCs (G-LCs) in the mesangium recognize their target structure and induce two distinct mesangiopathies, monoclonal Ig deposition disease (MIDD) and AL-amyloidosis (AL). Tubulopathic LCs (T-LCs) act on the proximal or on the distal tubule and cause, respectively, Fanconi syndrome (FS) and cast nephropathy. Pathogenic monoclonal Igs have the propensity to deposit in different renal parenchymal structures in extracellular sites, because of the transformation of soluble precursors in insoluble products. Evidence suggests that somatic mutations can destabilize the normal LCs globular soluble structure and this could be the major driving force for precipitation. Based on these features, MG can be classified as conformational and depositional diseases. Electronmicroscopy (EM) analysis of renal biopsies in MG patients with glomerular diseases distinguishes two morphological aspects. MIDD and a recently identified entity named proliferative glomerulonephritis (GN) with monoclonal IgG deposits are both characterized by non-organized granular electrondense deposits. AL, immunotactoid (IT) glomerulopathy and monoclonal cryoglobulinemia are, instead, characterized by organized deposits such as fibrils or microtubules. Tubular diseases in MG patients produce two different histological patterns. In FS, monoclonal Igs form crystals in the renal interstitium able to induce a local intense flogosis, while in cast nephropathy monoclonal Igs precipitate with Tamm Horsfall protein (THP) in the proximal tubular lumen and induce tubular obstruction. The different morphological aspects are unrelated to specific clinical manifestations, while renal biopsy can diagnose different entities that can respond to different therapeutical schedules. This reveals the importance of the renal biopsy in the clinical management of the renal pathology in plasma cells dyscrasias, mainly when supported by the most advanced techniques of immunoelectronmicroscopy and polymerase chain reaction (PCR)-mediated analysis. Further elucidation of the molecular events involved in the pathogenesis of the different forms of renal damage is needed to design new and more effective therapeutical strategies. In particular, urinary proteomics seem to be promising in this setting. PMID- 16358224 TI - Histology and immunohistology of IgA nephropathy. AB - IgA nephropathy is a histologically diverse glomerular disease characterized by mesangial or mesangial plus peripheral glomerular capillary immune complex deposits that contain IgA as the dominant or co-dominant immunoglobulin type. The most common histologic manifestation of IgA nephropathy is mesangial proliferative glomerulonephritis (GN), most often focal but not infrequently diffuse. However, the light microscopic appearance of IgA nephropathy spans the entire range from histologically normal to diffuse proliferative and crescentic glomerulonephritis, much as is the case with lupus nephritis. This review examines the histologic diversity as well as the immunohistologic features of IgA nephropathy. PMID- 16358225 TI - Co-localization and interaction of b0,+-type amino acid transporter 1 (BAT1) with caveolin-1 in rat kidney. AB - BACKGROUND: Cystinuria has been proposed as an inherited disease causing disorders in renal cystine and basic amino acid transport in the proximal tubules. Although cystinuria-related amino acid transporter gene related to b0,+ type amino acid transporter (rBAT1) and its substrate transport properties have been reported, the functional regulatory mechanisms remain to be elucidated. In this study, protein-protein interaction between rBAT1 and caveolin (Cav)-1 was investigated. METHODS: The renal distribution of rBAT1, rBAT and Cav-1 were demonstrated by employing reverse transcriptase polymerase chain reaction (RT PCR) and Western blot analysis. Co-localization of rBAT1 and Cav-1 was observed by immunocytochemistry in primary cultured renal proximal tubule-derived cells using a confocal microscope. This result was confirmed by Western blot analysis of isolated caveolae-rich membrane fraction and immunoprecipitation experiments using respective antibodies. RESULTS: In the separated rat kidney tissues following the corticomedullary axis, Cav-1 mRNA and protein expressions were increased from the cortex to the inner medulla. rBAT1 mRNA and protein expression were detected mainly in the outer medulla. Confocal microscopic results showed rBAT1 and Cav-1 co-localization in the plasma membrane. This result was confirmed by Western blot analysis of caveolae-rich membrane fraction and immunoprecipitates by respective antibodies. The effect of Cav-1 on rBAT1 function was evaluated using Cav-1 antisense oligodeoxynucleotide (ODN). The [14C] arginine uptake by rBAT1 was unchanged by the treatment with antisense ODN. CONCLUSIONS: From these results, rBAT1 and Cav-1 share a cellular expression in the segregated caveolae structure. As caveolae are rich in signaling molecules, BAT1 could play a role in diverse pathophysiological processes. PMID- 16358226 TI - "Point of no return (PNR)" in progressive IgA nephropathy: significance of blood pressure and proteinuria management up to PNR. AB - BACKGROUND: Based on observations of the clinical course in patients with IgA nephropathy (IgAN), D'Amico et al proposed the concept of the "point of no return (PNR)", after which progression to end-stage renal disease (ESRD) becomes inevitable. They reported that the approximate PNR is serum creatinine (sCr) 3.0 mg/dL. METHODS: To confirm the PNR and to clarify the factors affecting renal function deterioration in IgAN patients, we analyzed the sequential data of those with 1.2 or= 102 mmHg and/or urinary protein (UP) score >or= 2.0 with sCr up to 2.0 mg/dL was significantly poor. Multivariate analysis using the Cox's proportional hazards model, identified only MBP and UP during the course until sCr reached 2.0 mg/dl as independent prognostic factors for ESRD, having hazard ratios of 2.56 (per 10 mmHg; 95% confidence interval (95% CI) 1.08-6.05) and 4.37 (per 0.5 point; 95% CI 1.36-14.1), respectively. CONCLUSIONS: We confirmed PNR as a sCr level of 2.0 mg/dL (equivalent to estimated glomerular filtration rate (GFR) of 30-35 mL/min/1.73m2) during the course of IgAN in Japan. Proper management of both BP and UP until sCr has reached PNR is essential to arrest the progression to ESRD in IgAN. PMID- 16358227 TI - The prognostic value of renal biopsy in type 2 diabetes mellitus patients affected by diabetic glomerulosclerosis. AB - BACKGROUND: Although several studies have dealt with clinicopathological correlations in diabetic glomerulosclerosis (DGS), few have investigated the prognostic value of the renal biopsy. METHODS: One hundred and thirty-five type2 diabetes mellitus (DM) patients with bioptically proven DGS and a mean follow-up of 56.1 months were subdivided in 5 groups according to the outcome: 1) living with preserved renal function; 2) living with renal failure, not requiring dialysis; 3) living in dialysis; 4) dead in dialysis; 5) dead with preserved renal function. Duration of DM, creatininemia and proteinuria values at the time of biopsy and a histologic scoring (from 0 to 3) of 10 morphologic features were considered for statistical analysis. RESULTS: Statistically significant differences were observed in the distribution of the above mentioned parameters in the 5 groups with the exception of the duration of DM. The prognosis of DGS is poor: 79 patients needed dialysis and 60 died. Univariate analysis demonstrated the prognostic value of creatininemia (>or= 1.5 mg/dL), proteinuria (>or= 3 g/d) and histologic score (>or= 10) in assessing the relative risk of progression to dialysis (OR= 9.75, 4.12 and 11 respectively). The prognostic value of the histologic score (or= 2) was maintained when each morphologic parameter was separately evaluated (OR ranging from 2.8 to 8.5). Multivariate analysis was applied and only serum creatinine and histological score maintained their statistical significance (OR=4.45 and 2.46). The independence of these two variables means that the risk of progression to dialysis is multiplied in each single patient. CONCLUSIONS: Renal biopsy adds reliable information to that supplied by clinical and laboratory findings in DGS and therefore its extensive adoption should be recommended. Thanks to the prognostic value of the single morphologic parameters, also small tissue samples can give reliable results. PMID- 16358228 TI - Urinary sediment features in proliferative and non-proliferative glomerular diseases. AB - BACKGROUND: The studies on urine sediment particles in patients with glomerular diseases (GD) are few and have focused only on single urine particles. In this study, we investigated the prevalence and number of 12 urine sediment particles in two groups of patients, one with proliferative GD, and the other with non proliferative GD. METHODS: The urine sediment of 100 consecutive patients, with a renal biopsy-proven proliferative or non-proliferative GD and marked cylindruria, were examined a few hours before renal biopsy according to a standardized method. The urine particles investigated were erythrocytes, leukocytes, renal tubular cells, lipids and hyaline, hyaline-granular, granular, waxy, erythrocytic, leukocytic, epithelial and fatty casts. RESULTS: Patients with proliferative GD (n=52) had both a significantly higher prevalence of microscopic hematuria, leukocyturia, tubular epithelial cells, erythrocytic casts, epithelial casts, and significantly higher amounts of erythrocytes,leukocytes, tubular epithelial cells/20 high power field (HPF), erythrocytic and epithelial casts. On the other hand, patients with non-proliferative GD (n=48) had significantly higher numbers of fatty casts. In proliferative GD, leukocyturia was associated with intracapillary and extracapillary proliferation, crescents and fibrinoid necrosis at renal biopsy. At discriminant analysis, the two types of GD could be identified with 80.8% sensitivity and 79.2% specificity. By multiple logistic regression analysis, patients with erythrocytes, leukocytes and erythrocytic casts in the urine had an odds ratio (OR) of 9.91 (95% confidence interval (95% CI): 1.01-97.51), 7.85 (95% CI: 2.77-22.20), and 4.33 (95% CI: 1.41-13.31), respectively, of having proliferative GD. CONCLUSIONS: Our examination of the urine sediment shows that proliferative GD and non-proliferative GD differ in many respects. PMID- 16358229 TI - Treatment of steroid-resistant nephrotic syndrome with cyclosporine: study of 17 cases and a literature review. AB - BACKGROUND: Cyclosporine (CsA) has been used in steroid-resistant idiopathic nephrotic syndrome (INS) in many previous studies. OBJECTIVE: To evaluate if CsA is a therapeutic option for steroid-resistant nephrotic syndrome. METHODS: We performed a retrospective cohort study to evaluate the effects of CsA in 17 steroid-resistant INS patients. The main laboratorial data, before and after the use of CsA, and the response to CsA were evaluated. A literature review on this subject was also done. RESULTS: Patient age ranged from 2-43 yrs. Pre-treatment renal biopsy demonstrated focal segmental glomerulosclerosis (FSGS) (64%), membranous nephropathy (MGN) (12%), mesangial glomerulonephritis (MSGN) (12%) and minimal change disease (MCD) (12%). Pre-treatment laboratory tests showed a mean 24-hr proteinuria of 4372 +/- 2686 mg/dL. Treatment with CsA was given for a minimum of 3 months and a maximum of 98 months. Mean 24-hr proteinuria declined from 3181 +/- 2277 before CsA to 915 +/- 1140 mg/24 hr after CsA (p<0.001). Remission was seen in 70.5% of patients, being 52.9% complete and 17.6% partial. The adverse effects observed were nephrotoxicity (11.7%), hypertrichosis (5.8%) and gingival hyperplasia (5.8%). Relapses were seen in eight patients (47%), with posterior remission in six patients (75%). CONCLUSION: Data from the literature suggest that CsA is a good therapeutic option for patients with steroid-resistant INS, being effective in reducing proteinuria. The beneficial effect of CsA demonstrated in our study was limited due to its design and the small sample size. PMID- 16358230 TI - Endothelial function, CRP and oxidative stress in chronic kidney disease. AB - BACKGROUND: Chronic kidney disease (CKD) is associated with increased morbidity and mortality in cardiovascular disease (CVD). Apart from traditional risk factors, chronic inflammation, oxidative stress, malnutrition and endothelial dysfunction are important in CVD development in renal patients. Our aim was to investigate the relationship between high sensitivity C-reactive protein (CRP), endothelium dependent vasodilation (EDV) and oxidative stress markers in patients with CKD K/DOQI stage 3-5. METHODS: Measurements of CRP, conjugated dienes (CD), lipid hydroperoxide (LOOH), oxidized low density lipoprotein,glutathione and albumin were performed in 44 consecutive patients with CKD stage 3-5. EDV was measured by methacholine infusion in the brachial artery and venous occlusion plethysmography. RESULTS: Patients with high CRP had significantly lower glomerular filtration rates and albumin, but increased LOOH and CD. In multiple regression analysis, only LOOH and CD remained significant. Patients with poor EDV had increased urea and lower glutathione (GSH). In multiple regression analysis, GSH and urea were independently related to EDV. No correlation was found between CRP and endothelial function. CONCLUSION: CRP was related to lipid peroxidation, while endothelial function was related to intracellular oxidative stress in patients with CKD. CRP and EDV were unrelated to each other. Therefore, CRP and endothelial function could provide complementary prognostic information regarding future cardiovascular disorders in renal patients. PMID- 16358231 TI - Urotensin II in end-stage renal disease: an inverse correlate of sympathetic function and cardiac natriuretic peptides. AB - BACKGROUND: Urotensin II (UTN) is a peptide highly conserved across species with disparate effects on the vascular system and it is currently unclear whether high plasma UTN levels play a vasculotoxic or a vasculoprotective role. METHODS: In this study, we investigated the relationship between plasma UTN and sympathetic activity and cardiac natriuretic hormones in 191 hemodialysis (HD) patients without clinical evidence of heart failure. RESULTS: Plasma UTN was significantly higher in patients with end-stage renal disease (ESRD) (median: 6.5 ng/mL) than in age matched healthy subjects (median: 3.1 ng/mL) (p<0.001). On univariate analysis, UTN was inversely related to heart rate (r=-0.24), dialysis treatment duration (r=-0.27), norepinephrine (r=-0.28), neuropeptide Y (NPY) (r=-0.66), brain natriuretic peptide (BNP) (r=-0.41) and atrial natriuretic peptide (ANP) (r=-0.28) (all p<0.008). Of note, in multiple regression analyses these associations maintained strength similar to that of the corresponding unadjusted correlation coefficients. CONCLUSIONS: The inverse links between UTN and neuro hormonal factors indicate that UTN down-regulation in the presence of high sympathetic activity and high BNP could be a counter-regulatory response aimed at mitigating cardiovascular (CV) damage or that UTN itself acts as a protective factor. PMID- 16358232 TI - Relationship of p22phox C242T polymorphism with nephropathy in type 2 diabetic patients. AB - BACKGROUND: In this case-control study, we investigated the possible involvement of the p22phox C242T polymorphism in the development and progression of diabetic nephropathy (DN) in 535 Caucasian Brazilians with type 2 diabetes. We also evaluated the effects of the interaction of the C242T polymorphism with smoking and hypercholesterolemia on the susceptibility to nephropathy. METHODS: Genotype analysis was performed using polymerase chain reaction (PCR) followed by digestion with restriction enzyme. Logistic regression analysis was used to control for independent risk factors associated with nephropathy. RESULTS: The genotype frequencies in patients with overt DN (CC/CT/TT: 0.36/0.47/0.17) were not significantly different from those of diabetic individuals with normoalbuminuria (0.47/0.41/0.12) or microalbuminuria (0.42/0.48/0.10) (p=0.214). Likewise, there were no differences in the T allele frequency among patients with normoalbuminuria, microalbuminuria or overt DN (0.33, 0.34 and 0.40, respectively; p=0.111). However, the T allele was found to be more frequent among smokers with overt nephropathy (macroalbuminuria and/or in dialysis) than those who had normoalbuminuria (43 vs. 32%, p=0.045). The multiple logistic regression analysis confirmed that the CT+TT genotypes were independently associated with a higher risk of having overt nephropathy among smokers [odds ratio (OR)=6.76, 95% confidence interval (95% CI) 1.83-25.02]. CONCLUSIONS: Our study shows a gene environment interaction associated with the increased risk of DN progression in Caucasian Brazilian smokers with type 2 diabetes. Further studies should be performed to clarify whether it exists, and to what extent there is a relationship between the p22phox C242T polymorphism and DN. PMID- 16358233 TI - Assessment of phosphorus and calcium metabolism and its clinical management in hemodialysis patients in the community of Valencia. AB - AIM: The objective of the study was to determine the situation concerning mineral metabolism and bone disease in hemodialysis (HD) patients living in the community of Valencia (Spain), as well as the clinical practices for bone disease control in relation to the laboratory targets recommended in the National Kidney Foundation Dialysis Outcomes Quality Initiative (K/DOQI) guidelines. METHODS: In December 2003, a cross-sectional study was performed including 2392 patients (1485 males and 907 females) from 43 different centers in the council of Valencia (the entire HD population). Mean age was 65.8 +/- 14 yrs. Cut-off levels for the study of calcium, phosphorus, calcium-phosphorus product (Ca x P) and parathyroid hormone (PTH) were performed following the recommendations of the K/DOQI guidelines. RESULTS: The mean values for calcium were 9.57 +/- 0.7 mg/dL, phosphorus 4.97 +/- 1.5 mg/dL, intact PTH (iPTH) 297 +/- 353 pg/mL, Ca x P 47.5 +/- 15 mg2/dL2. Hypocalcemia (<8.4 mg/dL) was present in 5% of patients, whereas 17.8% of patients presented hypercalcemia (>10.2 mg/dL), 60.3% of whom received vitamin D. Hypophosphoremia (<3.5 mg/dL) was present in 16% of patients, and 29% of patients presented hyperphosphoremia (>5.5 mg/dL). Ca x P was <55 mg2/dL2 in 73% of patients. Thirty one percent of patients presented secondary hyperparathyroidism (HPTH >300 pg/mL), being severe in 12% (>600 pg/mL); 43% of patients presented iPTH <150 pg/mL. Only 7.3% of patients achieved the four recommendations provided in the K/DOQI guidelines. Vitamin D treatment was administered in 48% of patients. CONCLUSIONS: The population undergoing dialysis in the community of Valencia achieved targets based on the clinical recommendations of the K/DOQI guidelines as follows: 45% of patients achieved targets for calcium, 55% for phosphorus, 73% for Ca x P and 26% for iPTH levels. Surprisingly, only 7.3% of patients achieved all four targets. PMID- 16358234 TI - Stenting of renal artery stenosis: is it beneficial in chronic renal failure? AB - BACKGROUND: The utility of renal artery stenting (RS) in preserving renal function (RF) is not well established. Our prospective study aimed to examine the clinical effects of RS in patients with proximal renal artery stenosis and chronic renal failure (CRF). METHODS: Fifty-two patients, with atherosclerotic renal artery stenosis (ARAS) and renal impairment underwent successful monolateral (33 patients) or global RS (19 patients: six bilateral stenting, two surgical solitary kidney and 11 functional solitary kidney). Patients were considered eligible if they had at least a mild renal impairment (serum creatinine (Cr) >1.5 mg/dL). To assess a significant change in RF, we compared the slopes of the regression lines derived from the reciprocal of Cr vs. time, plotted before and after stenting. Patients had a median post-procedure follow-up of 24 months (range 9-54). RESULTS: Before stenting all patients exhibited a negative slope, indicating progressive renal failure. After stenting the slopes became positive in eight patients (15.5%), indicating a significant improvement in RF; in 31 patients (59.5%) the slopes were not significantly different from 0 and were associated with a stable RF, while 13 patients (25%) presented negative slopes and showed a continuous reduction in RF. Previous serum Cr, initial diameter of the treated kidney, vascular resistive index, and mono vs. global stenting were not significantly associated to post stenting RF. CONCLUSIONS: RS appears to be associated with RF stabilization in the majority of patients with CRF. PMID- 16358235 TI - The impact of catheter-restricted filling with cefotaxime and heparin on the lifespan of temporary hemodialysis catheters: a case controlled study. AB - BACKGROUND: Reduction in the rates of major complications such as infection and thrombosis that limit the lifespan of hemodialysis (HD) catheters could conceivably lead to improved survival of "temporary" non-tunneled HD catheters (NTCs). This study was designed to evaluate the impact of the "locking"' of a broad-spectrum antibiotic-cefotaxime with heparin, on the incidence of catheter thrombosis, catheter-related bloodstream infections (CRBSI) and the NTC lifespan. METHODS: This prospective study included 208 (109 males and 99 females) end-stage renal disease (ESRD) patients of diverse etiology enrolled for long-term HD from July 2002 to June 2003 at our tertiary care hospital. Those patients requiring NTC insertion for the maintenance or commencement of HD were eligible for the study. Cefotaxime-heparin "lock" solutions were freshly prepared by dissolving sterile cefotaxime sodium powder for injection directly in the heparin sodium to reach a concentration of 10 mg/mL for cefotaxime and 5000 U/mL for heparin. A final volume of 5 mL was prepared in a syringe using aseptic precautions to fill 1.3 mL in the venous and 1.2 mL in the arterial lumen of the catheter with a combined volume of approximately 2.5 mL. Blood samples were collected for culture and sensitivity, simultaneously, from the catheter hub and the peripheral vein, on clinical suspicion of CRBSI. Thrombosis was defined as the inability to use the catheter at a blood flow of 200 ml/min, which did not respond to catheter repositioning and/or intraluminal thrombolysis. RESULTS: An overall relative risk reduction (RRR) of 56.5% for catheter thrombosis and separately for all types of NTCs (femoral (FC) - 52.7%, subclavian (SC) - 55.9%, and internal jugular (IJC) - 53.7%), were observed in group II having a cefotaxime-heparin lock solution. A lower CRBSI incidence (1.65 vs. 3.13/1000 catheter days) compared with group I, leading to a RRR of 50.5% was also recorded in group II. Significantly higher percentage catheter survival rates at 7, 14 and at 28 days for FC and at 14, 28 and 56 days for SC and IJC were observed among patients in group II in comparison to group I. CONCLUSIONS: Cefotaxime-heparin locks led to a significant reduction in catheter thrombosis and CRBSI incidence. The enhanced NTC lifespan thus achieved could help physicians in better assessment of the patient's vasculature prior to the placement of permanent vascular access. PMID- 16358236 TI - Fibrinogen and calcium x phosphate product are predictors of cardiovascular events in the CAPD population. AB - OBJECTIVE: Plasma fibrinogen (Fib) and calcium x phosphate product have emerged as cardiovascular (CV) event predictors in hemodialysis (HD) patients, but their role in peritoneal dialysis (PD) is less studied. DESIGN AND SUBJECTS: We investigated whether Fib and calcium x phosphate product predict CV events in a prospective cohort study of 47 continuous ambulatory PD (CAPD) patients (mean follow-up 34.6 months). RESULTS: During the follow-up, 29 patients experienced CV events, which were fatal in 11 of them. Plasma Fib was markedly elevated in the entire population studied (median 612 mg/dL inter-quartile range (IQ): 566-718 mg/dL). On univariate analysis, Fib was higher (p=0.02) in those patients who had fatal or non-fatal CV events (median 654 mg/dL, IQ: 577-801) than in event-free patients (median 579 mg/dL, IQ: 532-629). Patients with incident CV events also tended to have higher calcium x phosphate product (51.6 +/- 12.8 vs. 43.8 +/- 13.9 mg2/dL2, p=0.06). On multivariate Cox's regression analyses, including traditional risk factors and history of previous CV events, Fib (hazard ratio (HR) associated with 100 mg/dL increase in plasma Fib 1.29 95% confidence interval (CI) 1.03-1.63 (p=0.03)) and calcium x phosphate product (HR associated with a 5 mg2/dL2 increase 1.25 95% CI 1.05-1.49 (p=0.01)) emerged as independent CV event predictors. CONCLUSIONS: Calcium x phosphate product and Fib are CV event predictors in CAPD patients. PMID- 16358237 TI - Anderson-Fabry disease: a protean clinical behavior and a chance diagnosis. AB - Anderson-Fabry disease is a rare inborn X-linked glycosphingolipid storage disorder in which the deficient activity of the enzyme alfa-galactosidase A (alfa gal A) leads to the progressive tissular accumulation of lipidic molecules which, in turn, cause a protean pattern of multi-organ disfunction. Enzyme replacement therapy has recently become available and has proved to be effective in controlling the disorder. We present and discuss the case of a family with this disease, with special attention to the variability of clinical features and the difficulty of a correct diagnosis. PMID- 16358238 TI - Association of IgA nephropathy, hypothyroidism and hypercholesterolemia. AB - Immunoglobulin A (IgA) nephropathy can rarely be associated with thyroid disease. We present a case of a young lady with nephrotic range proteinuria, microscopic hematuria, a creatinine clearance of 67 ml/min, biopsy proven IgA nephropathy, in whom hypercholesterolemia persisted after immunosuppressive therapy induced remission of glomerulonephritis. Fluvastatin was given but rhabdomyolysis developed. Unexpectedly, CK remained elevated following discontinuation of fluvastatin. Secondary amenorrhoea prompted endocrine work-up and hypothyroidism was diagnosed. Cholesterol, CK and TSH values became normal within 3 months on L thyroxin therapy. The literature of the association of glomerular diseases and thyroid illness is reviewed. We would like to draw the attention to the possible association of glomerular pathologies and thyroid diseases and the importance of ruling out hypothyroidism and measuring CK level before starting statin therapy. PMID- 16358239 TI - Nephrotic syndrome revealing a disseminated Langerhans' cell histiocytosis. AB - Langerhans' cell histiocytosis (LCH) is a clonal disorder of proliferating histiocytes, which can affect various organs. The clinical picture ranges from localized to disseminated disease with multiple organ involvement and severe organ dysfunction. Renal involvement is uncommon and the association with glomerulonephritis is unusual. We report here a case of nephrotic syndrome caused by membranoproliferative glomerulonephritis (MPGN) in a 20-year-old woman, which revealed a disseminated form of LCH. Surgical removal of the LCH process and high doses of corticosteroids led to the remission of the basic disease and the renal involvement. The occurence of membranoproliferative glomerulonephritis in the course of LCH might be mediated by circulating immune complexes, composed at least in part of antigens or neoantigenes which may cause the abnormal proliferation of the Langerhans' cells. PMID- 16358240 TI - A rare post-transplant malignancy, cerebellar hemangioblastoma: a case report. AB - INTRODUCTION: Post-transplant malignancies are among the most important complications in organ transplantation. Hemangioblastoma (HB) is especially prevalent in the cerebellum. CASE REPORT: A 20-year-old male who first started dialysis therapy, and then underwent kidney transplantation from a living relative donor. Five years after transplantation, the patient suffered from vertigo and imbalance when walking. On cranial magnetic resonance imaging (MRI), a mass lesion in the right cerebellar hemisphere was observed, 3 x 3 x 3 cm in size, which was pushing against the fourth ventricle, and the right cerebellar peduncle. The patient had significant hydrocephaly. The mass lesion was removed by craniectomy. The pathological diagnosis was cerebellar hemangioblastoma. The symptoms and clinical findings improved. The patient was diagnosed with sporadic hemangioblastoma. Rapamycin therapy was started instead of cyclosporine, and the patient is being followed up without further problems. DISCUSSION: HB causes 2% of all intracranial tumors in the general population. It is generally sporadic in nature and approximately 20% can be associated with von Hippel-Lindau (VHL) syndrome. As in this case, MRI is preferred for the diagnosis. There was no pathology related to VHL disease in this patient's physical examination, family history, routine biochemical tests and abdominal MRI. The treatment is surgical excision of the tumor, as in this case. CONCLUSION: When cerebellar symptoms occur or a cerebellar mass lesion is detected in an organ recipient, HB should be considered in the differential diagnosis. The examination of patients with HB for a possible association with VHL disease is also required. PMID- 16358241 TI - Graft mucormycosis in a renal allograft recipient. AB - Mucormycosis is a rare but potentially lethal fungal infection in the renal allograft recipient. Renal allograft mucormycosis is extremely rare. To date, only five cases have been reported in the literature. We report a case of renal allograft mucormycosis, where this complication developed 18 months after transplantation following treatment for acute rejection with high dose steroids and antithymocyte globulin. PMID- 16358242 TI - Fatal Strongyloides stercoralis infection in a young woman with lupus glomerulonephritis. AB - We report a case of fatal Strongyloides stercoralis (S. stercoralis) infection in a 35-year-old female with lupus glomerulonephritis after prolonged steroid therapy. An epidemiological evaluation revealed that our patient originated from a rural area in Bosnia and Herzegovina, which was an endemic region of S. stercoralis infection in the former Yugoslavia. She had severe gastrointestinal and pulmonary symptoms and history of a 13 kg weight loss in 3 months. Histopathological examination of large bowel mucosa showed nematode larvae. Microscopy of stool, sputum and urine samples confirmed S. stercoralis. The diagnosis was delayed because of the low suspicion index, the absence of eosinophilia and non-specific signs of infection, which could be a result of the underlying disease or the effects of corticosteroids. This case highlights the importance of screening for S. stercoralis in patients starting immunosuppressive therapy, especially if they are from endemic areas. Timely diagnosis and therapy could save the patient's life. PMID- 16358243 TI - Pasteurella multocida peritoneal dialysis-associated peritonitis: a report of two cases and review of the literature. AB - We present 2 new cases of pasteurella multocida induced peritonitis in patients on long term CCPD. We also briefly review the literature on a total of 16 cases (including the present 2 cases) reported in the English literature. PMID- 16358244 TI - Idiopathic retroperitoneal fibrosis. PMID- 16358245 TI - Monitoring redox-dependent contribution of lipids in Fourier transform infrared difference spectra of complex I from Escherichia coli. AB - Biochemical and crystallographic studies have shown that phospholipids are essential for the integrity and activity of membrane proteins. In the study presented here, we use electrochemically induced Fourier transform infrared (FTIR) spectroscopy to demonstrate variations occurring upon the presence and absence of lipids in NADH:ubiquinone oxidoreductase (complex I) from Escherchia coli by following the C=O vibration of the lipid molecule. Complex I is activated in the presence of lipids. Interestingly, in electrochemically induced FTIR difference spectra of complex I from E. coli, a new signal at 1744/1730 cm(-1) appears after addition of E. coli polar lipids, concomitant with the oxidized or reduced form, respectively. Absorbance spectra of liposomes from mixed lipids at different pH values demonstrate shifts for the carbonyl vibration depending on the environment. On this basis we suggest that lipids, though not redox active themselves, contribute in reaction-induced FTIR difference spectra, if a change occurs in the direct environment of the lipid during the observed reaction or coupled processes. PMID- 16358246 TI - Inhibition and partial reactions of Na,K-ATPase studied by Fourier transform infrared difference spectroscopy. AB - Reaction-induced infrared (IR) difference spectroscopy with caged ATP and Na,K ATPase allows us to differentiate unambiguously between phosphorylated and unphosphorylated states of the enzyme as well as of its ouabain complex. The IR spectral changes upon phosphoenzyme formation are characterized and interpreted. Our results show clearly that high Na(+) concentrations prevent the binding of ouabain with high affinity, which is consistent with the results of a corresponding kinetic study employing spectrofluorimetry and calorimetric titrations. This unexpected antagonism leading to low ouabain affinity is assumed related to a conformation of the protein, induced by low affinity binding of the third Na(+) ion. We thus conclude that not the free enzyme but a phosphorylated state of the reaction cycle preferentially binds ouabain and leads to the loss of hydrolytic activity. PMID- 16358247 TI - DNA compaction into new DNA vectors based on cyclodextrin polymer: surface enhanced Raman spectroscopy characterization. AB - The ability of DNA to bind polycation yielding polyplexes is widely used in nonviral gene delivery. The aim of the present study was to evaluate the DNA compaction with a new DNA vector using Raman spectroscopy. The polyplexes result from an association of a beta-cyclodextrin polymer (polybeta-CD), an amphiphilic cationic connector (DC-Chol or adamantane derivative Ada2), and DNA. The charge of the polymeric vector is effectively controlled by simple addition of cationic connector in the medium. We used surface enhanced Raman spectroscopy (SERS) to characterize this ternary complex, monitoring the accessibility of adenyl residues to silver colloids. The first experiments were performed using model systems based on polyA (polyadenosine monophosphate) well characterized by SERS. This model was then extended to plasmid DNA to study polybeta-CD/Ada2/DNA and polybeta-CD/DC-Chol/DNA polyplexes. The SERS spectra show a decrease of signal intensity when the vector/DNA charge ratio (Z+/-) increases. At the highest ratio (Z+/- = 10) the signal is 6-fold and 3-fold less intense than the DNA reference signal for Ada2 and DC-Chol polyplexes, respectively. Thus adenyl residues have a reduced accessibility as DNA is bound to the vector. Moreover, the SERS intensity variations are in agreement with gel electrophoresis and zeta potential experiments on the same systems. The overall study clearly demonstrates that the cationic charges neutralizing the negative charges of DNA result in the formation of stable polyplexes. In vitro transfection efficiency of those DNA vectors are also presented and compared to the classical DC-Chol lipoplexes (DC-Chol/DNA). The results show an increase of the transfection efficiency 2-fold higher with our vector based on polybeta-CD. PMID- 16358248 TI - The effect of water on the glass transition of human hair. AB - The glass transition of human hair and its dependence on water content were determined by means of differential scanning calorimetry (DSC). The relationship between the data is suitably described by the Fox equation, yielding for human hair a glass transition temperature of T(g) = 144 degrees C, which is substantially lower than that for wool (174 degrees C). This effect is attributed to a higher fraction of hydrophobic proteins in the matrix of human hair, which acts as an internal plasticizer. The applicability of the Fox equation for hair as well as for wool implies that water is homogeneously distributed in alpha keratins, despite their complex morphological, semicrystalline structure. To investigate this aspect, hair was rendered amorphous by thermal denaturation. For the amorphous hair neither the water content nor T(g) were changed compared to the native state. These results provide strong support for the theory of a quasi homogeneous distribution of water within alpha-keratins. PMID- 16358250 TI - Multiplex multidimensional nanoLC-MS system for targeted proteomic analyses. AB - The present work describes a dual-column and dual-sprayer LC-MS system for high throughput proteomic analyses. This system consists of two precolumns for sample desalting and two analytical columns. Each column is terminated by a nanoelectrospray emitter mounted on a robotic arm enabling their sequential positioning in front of the sampling cone of the mass spectrometer. The effluent from each emitter is recorded in separate acquisition channels without detectable crosstalk. Gradient elution to both nanoLC columns is delivered by a single HPLC system via a flow splitter. The reproducibility of retention time and peak intensity of the present multiplex system were comparable to those obtainable using a single emitter configuration. Replicate injections of complex tryptic digests (n = 10) indicated that this system provided good reproducibility of retention time and peak intensity on both columns with RSD values of less than 0.9 and 18.6%, respectively. The application of this system is demonstrated for the monitoring of protein expression changes in U937 human monocyte cells with and without phorbol ester administration. Furthermore, we also demonstrated the use of this multiplex system in a 2-D LC configuration to increase sample loading and throughput for the analysis of biomarker samples of higher complexity. Variations in peptide abundance down to two-fold change were identified across salt fractions for spiked tryptic digests present at a level of 50 fmol in 1.5 microg of plasma samples. PMID- 16358251 TI - Active mixing inside microchannels utilizing dynamic variation of gradient zeta potentials. AB - This study presents a new active micromixer with high mixing efficiency achieved by means of a gradient distribution of the surface zeta potential controlled by changing the frequency of voltage applied on shielding electrodes. Gradient surface zeta potential is generated by applying a high voltage to inclined buried shielding electrodes. While alternating the frequency of driving voltage, the zeta potential could be changed accordingly, thus providing a significant mixing effect inside microchannels. A theoretical model is proposed to predict the distribution of zeta potential. The results from this model are critically compared with the well-developed three-capacitor model. Additionally, two time factor scales, the charge time of capacitor and mixing length flow time, are used to predict the optimum frequency. The prediction of optimum frequency, 0.5 Hz, is consistent with experimental results. Moreover, a five-pair inclined shielding electrode with a frequency of 0.5 Hz leads to a significant improvement in the mixing performance of the active micromixer. Numerical results indicate that a localized flow circulation is generated when the control voltage is applied to the inclined shielding electrodes. Furthermore, the streamlines are experimentally observed by using fluorescent beads. The shape of this circulation is dependent on the distribution of gradient zeta potential, which is determined by the arrangement of electrodes. The effects of the number of electrode pairs and the layout of shielding electrodes on the mixing performance of micromixer are also explored both numerically and experimentally. It is revealed that five pair inclined electrodes at 0.5 Hz provide the highest mixing efficiency. Finally, a reaction between N-benzoyl-L-arginine-p-nitroanilide and trypsin enzyme is performed to verify the capability of micromixers. The experimental results reveal that the reaction can achieve a higher performance indicating a higher mixing efficiency. The active micromixers could be used in microfluidic systems for improving the mixing efficiency and thus enhancing the bioreaction. PMID- 16358252 TI - On the surface modification of microchannels for microcapillary electrophoresis chips. AB - This paper presents systematic investigation of the microchannel surface properties in microCE chips. Three popular materials for microCE chips, polydimethylsiloxane (PDMS), quartz, and glass, are used. The zeta potentials of these microchannels are calculated by measuring the EOF velocity to evaluate the surface properties after surface modification. The hydrophobic PDMS is usually plasma-treated for microCE applications. In this study, a new method using a high throughput atmospheric plasma generator is adopted to treat the PDMS surface under atmospheric conditions. In this approach, the cost and time for surface treatment can be significantly reduced compared with the conventional vacuum plasma generator method. Experimental results indicate that new functional groups could be formed on the PDMS surface after treatment, resulting in a change in the surface property. The time-dependent surface property of the plasma-treated PDMS is then measured in terms of the zeta potential. Results show that the surface property will reach a stable condition after 1 h of plasma treatment. For glass CE chips, two new methods for changing the microchannel surface properties are developed. Instead of using complicated and time-consuming chemical silanization procedures for CE channel surface modification, two simple and reliable methods utilizing organic-based spin-on-glass and water-soluble acrylic resin are reported. The proposed method provides a fast batch process for controlling the surface properties of glass-based CE channels. The proposed methods are evaluated using PhiX-174 DNA maker separation. The experimental data show that the surface property is modified and separation efficiency greatly improved. In addition, the long-term stability of both coatings is verified in this study. The methods proposed in this study show potential as an excellent solution for glass-based microCE chip surface modification. PMID- 16358253 TI - Static adsorptive coating of poly(methyl methacrylate) microfluidic chips for extended usage in DNA separations. AB - A simple and robust static adsorptive (dynamic) coating process using 2% hydroxyethylcellulose was developed for surface modification of poly(methyl methacrylate) (PMMA) microfluidic chips for DNA separations, suitable for usage over extended periods, involving hundreds of runs. The coating medium was also used as a sieving matrix for the DNA separations following the coating process. Four consecutive static treatments, by simply filling the PMMA chip channels with sieving matrix once every day, were required for obtaining a stable coating and optimum performance. The performance of the coated chips at different phases of the coating process was studied by consecutive gel electrophoretic separations with LIF detection using a PhiX-174/HaeIII DNA digest sample. The coated chip, with daily renewal of the sieving matrix, showed high stability in performance during a 25-day period of systematic study, involving more than 100 individual runs. The performance of the coated chip also remained almost the same after 3 months of continuous usage, during which over 200 separations were performed. The average precision of migration time for the 603-bp fragment was 1.31% RSD (n = 6) during the 25-day study, with a separation efficiency of 6.5 x 10(4) plates (effective separation length 5.4 cm). PMID- 16358254 TI - Direct coupling of polymer-based microchip electrophoresis to online MALDI-MS using a rotating ball inlet. AB - We report on the coupling of a polymer-based microfluidic chip to a MALDI-TOF MS using a rotating ball interface. The microfluidic chips were fabricated by micromilling a mold insert into a brass plate, which was then used for replicating polymer microparts via hot embossing. Assembly of the chip was accomplished by thermally annealing a cover slip to the embossed substrate to enclose the channels. The linear separation channel was 50 microm wide, 100 microm deep, and possessed an 8 cm effective length separation channel with a double-T injector (V(inj) = 10 nL). The exit of the separation channel was machined to allow direct contact deposition of effluent onto a specially constructed rotating ball inlet to the mass spectrometer. Matrix addition was accomplished in-line on the surface of the ball. The coupling utilized the ball as the cathode transfer electrode to transport sample into the vacuum for desorption with a 355 nm Nd:YAG laser and analyzed on a TOF mass spectrometer. The ball was cleaned online after every rotation. The ability to couple poly(methylmethacrylate) microchip electrophoresis devices for the separation of peptides and peptide fragments produced from a protein digest with subsequent online MALDI MS detection was demonstrated. PMID- 16358255 TI - A concept for miniaturized 3-D cell culture using an extracellular matrix gel. AB - This paper presents a novel method to embed, anchor, and cultivate cells in a controlled 3-D flow-through microenvironment. This is realized using an etched silicon pillar flow chamber filled with extracellular matrix (ECM) gel mixed with cells. At 4 degrees C, while in liquid form, ECM gel is mixed with cells and injected into the chamber. Raising the temperature to 37 degrees C results in a gel, with cells embedded. The silicon pillars both stabilize and increase the surface to volume ratio of the gel. During polymerization the gel shrinks, thus creating channels, which enables perfusion through the chip. The pillars increase the mechanical stability of the gel permitting high surface flow rates without surface modifications. Within the structure cells were still viable and proliferating after 6 days of cultivation. Our method thus makes it possible to perform medium- to long-term cultivation of cells in a controlled 3-D environment. This concept opens possibilities to perform studies of cells in a more physiological environment compared to traditional 2-D cultures on flat substrates. PMID- 16358258 TI - Quantitative analysis of DNA-porphyrin interactions. AB - The binding of manganese(III)-tetra(4-N-methylpyridyl)porphyrin (MnTMpyP) with synthetic poly(dA-dT)2, poly(dI-dC)2, and poly(dG-dC)2 DNAs as well as calf thymus (CT) DNA has been quantitatively studied in detail using induced CD (circular dichroism) spectroscopy in the Soret absorption band. The CD spectra, which changed greatly depending on the porphyrin to DNA base-pair molar ratio (r), were normalized with respect to DNA concentration and deconvoluted. Three independent component binding modes (named mode 1, 2, and 3 in the order of increasing r values) were identified, which successfully simulated the observed CD spectra with negligibly small residuals for a wide range of r values. In the case of poly(dA-dT)2, poly (dI-dC)2, and CT DNA, all the three modes appeared, whereas in the case of poly(dG-dC)2 DNA, only modes 1 and 3 appeared in the r range studied. The r dependence of each binding mode, i.e., its relative affinity toward DNA, has been revealed by this analysis. Mode 1, which appeared as a single binding mode at very low r values (r < or = ca. 0.05), was inhibited by the addition of methyl green, a drug that preferentially binds to the major groove of poly (dA-dT)2 DNA. Berenil, a known minor groove binder to poly(dA-dT)2 or poly(dI-dC)2 DNA, inhibited modes 2 and 3. From these inhibition experiments as well as comparison of the component spectra for DNAs of different sequence, a binding site on DNA was proposed for each component binding mode. The number of DNA base pairs covered by a single molecule of porphyrin was estimated. PMID- 16358260 TI - Epidemiology and the causes of breast cancer. AB - This article describes the characteristics of 3 dominant features of breast cancer epidemiology. These characteristics include the association of disease risk with childbearing, its relationship to ovarian activity and its international variation (particularly as the latter differs in the years before and after menopause). Equivocal tests of one hypothesis that reconciled some of these features through variations in levels of the fractions of estrogen are described. Other hypotheses with a similar objective are needed. The 3 known causes of human breast cancer, ionizing radiation, exogenous ovarian hormones and beverage alcohol, offer some preventive possibilities but do little to explain the epidemiologic features of the majority of cases of the disease that occur in their absence. PMID- 16358261 TI - Functional variants in the promoter of interleukin-1beta are associated with an increased risk of breast cancer: a case-control analysis in a Chinese population. AB - Interleukin 1beta (IL-1beta) is a multifunctional cytokine that upregulates the inflammatory response, and participates in carcinogenesis, malignant transformation, tumor growth, invasion and metastasis. Two potentially functional polymorphisms (T-31C and C-511T) in the IL-1beta gene promoter were suggested to be correlated with alteration of IL-1beta expression and therefore may be associated with cancer risk. To test the hypothesis that these 2 polymorphisms are associated with risk of breast cancer, we performed a case-control study of 365 breast cancer cases, 270 patients with benign breast diseases (BBD) and 631 cancer-free controls in a Chinese population. Multivariate logistic regression analyses revealed that increased risk of breast cancer was associated with IL 1beta-31C variant genotypes [adjusted odds ratio (OR)=1.28 and 95% confidence interval (CI)=0.91-1.80 for -31CT and 1.72 (95% CI=1.16-2.54) for -31CC], compared with the -31TT genotype. Similarly, IL-1beta-511T variant genotypes were also associated with increased risk of breast cancer (adjusted OR=1.20, 95% CI=0.86-1.67 for -511CT and adjusted OR=1.74, 95% CI=1.18-2.56 for -511TT), compared with the -511CC genotype. Furthermore, cancer risks associated with IL 1betaT-31C variant genotypes were more evident in older women, postmenopausal women and individuals with a later menarche age. Interestingly, although we did not find significant associations of these 2 variants with cancer risk when compared with the BBD patients, a 1.27-fold (95% CI=1.01-1.60) increased risk was observed with the -31C-511T common haplotype. These findings indicate that these 2 IL-1beta promoter variants may contribute to risk of developing breast cancer in the Chinese population. PMID- 16358262 TI - Intracranial therapy of glioblastoma with the fusion protein DTIL13 in immunodeficient mice. AB - A fusion protein consisting of human interleukin-13 and the first 389 amino acids of diphtheria toxin was assembled in order to target human glioblastoma cell lines in a murine intracranial model. In vitro studies to determine specificity indicated that the protein called DTIL13 was highly selective for human glioblastoma. In vivo, the maximum tolerated dose of DTIL13 was 1 microg/injection given every other day and repeated for 3 days. Doses that exceeded this amount resulted in weight loss and liver damage as determined by histology and enzyme assay. Experiments in IL-4 receptor knockout mice revealed that liver toxicity was receptor-related. This same dose given to nude mice with established U373 MG brain tumors resulted in significant reductions in tumor volume and significantly prolonged survival (p<0.0001). Magnetic resonance imaging (MRI) proved to be extremely useful in (i) determining the ability of DTIL13 to reduce tumor size and (ii) for studying toxicity since diffusion weighted and gradient echo-weighted MRI revealed that vascular leak syndrome was not a limiting toxicity at this dose. These results suggest that DTIL13 is as effective in an intracranial rodent model as it was in a flank model in previous studies and that DTIL13 might be an effective treatment for glioblastoma multiforme. PMID- 16358263 TI - Neural crest cells and patterning of the mammalian dentition. AB - The mammalian dentition is composed of serial groups of teeth, each with a distinctive crown and root morphology, highly adapted to its particular masticatory function. In the embryo, generation of individual teeth within the jaws relies upon interactions between ectoderm of the first branchial arch and the neural crest-derived ectomesenchymal cells that migrate into this region from their site of origin along the neural axis. Classic tissue recombination experiments have provided evidence of an essential role of the ectoderm in initiating tooth development; however, the underlying ectomesenchyme rapidly acquires dominance in establishing shape. A key question is how these cells acquire this positional information. One theory suggests that ectomesenchymal cells are pre-patterned with respect to shape generation. Alternatively, this cell population acquires positional information within the first branchial arch itself, following migration. Recent molecular evidence suggests a high degree of plasticity within these ectomesenchymal cells. In particular, signalling molecules within the ectoderm exert a time-dependent influence upon the ectomesenchyme by establishing specific domains of homeobox gene expression. Initially, these ectomesenchymal cells are plastic and able to respond to signalling from the ectoderm, however, this plasticity is rapidly lost and pattern information becomes fixed. Therefore, in the first branchial arch, local regulation between the ectoderm and neural crest-derived ectomesenchyme is crucial in establishing the appropriate tooth shape in the correct region of the jaw. PMID- 16358264 TI - Heritability and heterochrony of polychromatism in a Lake Victoria Cichlid fish: Stepping stones for speciation? AB - In many haplochromine cichlid fish, male nuptial coloration is subject to female mate choice and plays a central role in the evolution of reproductive isolation between incipient species. Intraspecific variation in male coloration may serve as a target for diversifying sexual selection and provide a starting point for species divergence. Here, we investigated a polychromatism in Neochromis omnicaeruleus, a haplochromine from Lake Victoria, East-Africa. In this species, male coloration ranges from skyblue to yellow-red and females are grey-blue to yellow. We found that both genetic and environmental factors influence the expression of these colours during individual development. In a natural population, we found that male colour was associated with size and sexual maturity: yellow males were smaller than blue males and tended to be sexually immature. In females, size and maturity did not differ between colour types. Laboratory crosses revealed that there is a heritable component to the observed colour variation: yellow parents produced more yellow offspring than blue parents. Together with repeated aquarium observations of yellow individuals that gradually become blue, these data suggest that yellow males change to blue as they approach sexual maturity, and that the occurrence and timing of this transition is influenced by both environmental and genetic effects. The significance of this mechanism of colour expression as a possible target for divergent selection remains to be evaluated. PMID- 16358265 TI - Evolutionary genetics of vertebrate tissue mineralization: the origin and evolution of the secretory calcium-binding phosphoprotein family. AB - Three principal mineralized tissues are present in teeth; a highly mineralized surface layer (enamel or enameloid), body dentin, and basal bone. Similar tissues have been identified in the dermal skeleton of Paleozoic jawless vertebrates, suggesting their ancient origin. These dental tissues form on protein matrix and their mineralization is controlled by distinctive proteins. We have shown that many secretory calcium-binding phosphoproteins (SCPPs) are involved in tetrapod tissue mineralization. These SCPPs all originated from the common ancestral gene SPARCL1 (secreted protein, acidic, cysteine-rich like 1) that initially arose from SPARC. The SCPP family also includes a bird eggshell matrix protein, mammalian milk casein, and salivary proteins. The eggshell SCPP plays crucial roles in rigid eggshell production, milk SCPPs in efficient lactation and in the evolution of complex dentition, and salivary SCPPs in maintaining tooth integrity. A comparative analysis of the mammalian, avian, and amphibian genomes revealed a tandem duplication history of the SCPP genes in tetrapods. Although these tetrapod SCPP genes are fewer in teleost genomes, independent parallel duplication has created distinct SCPP genes in this lineage. These teleost SCPPs are also used for enameloid and dentin mineralization, implying essential roles of SCPPs for dental tissue mineralization in osteichthyans. However, the SCPPs used for tetrapod enamel and teleost enameloid, as well as tetrapod dentin and teleost dentin, are all different. Thus, the evolution of vertebrate mineralized tissues seems to be explained by phenogenetic drift: while mineralized tissues are retained during vertebrate evolution, the underlying genetic basis has extensively drifted. PMID- 16358267 TI - Effect of temporal synergism of neural oscillations on photorefractoriness in Japanese quail (Coturnix coturnix japonica). AB - Circadian rhythms in many metabolic functions including neural (transmitters) and hormonal secretion appear to change with physiological condition. It is also reported that seasonal changes in photoperiodism/reproduction and other metabolic conditions may result from a temporal interaction of circadian neural oscillations that change seasonally. To test this hypothesis, the present study was designed to study the effect of temporal synergism of two neural oscillations (serotonin and dopamine) on relative photorefractoriness of Japanese quail. Serotonin and dopamine precursor drugs (5-HTP, 5-hydroxytryptophan and L-DOPA, L dihydroxyphenylalanine) were administered (intraperitonially 5 mg/100 g body weight) at six different time intervals of 0, 4, 8, 12, 16 and 20 hr in sexually mature quail (>12 weeks old). The birds of control group received two daily injections of normal saline. The treatment was given for 13 days in continuous condition of light and then the quail were shifted to intermediate daylength (LD 13.5:10.5 for experiment 1) and short daylength (LD 8:16 for experiment 2). Six weeks following treatment, birds in intermediate daylength showed regressed cloacal gland and testicular activity except in 12-hr group, which exhibited gonadostimulatory condition. But birds of all the groups in short daylength showed complete regression of cloacal gland after 4 weeks of the treatment. In experiment 3, reproductively quiescent relative photorefractory quail maintained under intermediate daylength (LD 13.5:10.5) received 13 daily injections of 5-HTP and L-DOPA at the interval of 12 hr. At 6 weeks post-treatment, it was observed that unlike cloacal gland of control quail, which remained regressed, that of 12 hr quail showed significant development. These findings indicate that 12-hr temporal interaction of 5-HTP and L-DOPA administration maintained reproductive system in stimulated condition and prevented reproductive regression in photorefractory quail, but did not prevent the onset of scotosensitivity. It is concluded that the 12-hr temporal relationship of circadian serotonergic and dopaminergic oscillations not only eliminates photorefractoriness but may also re establish photosensitivity in relative photorefractory quail. These findings suggest the regulatory role of neural oscillations and their temporal interaction in the regulation of neuroendocrine-gonadal axis with special reference to photosensitivity/refractoriness. PMID- 16358268 TI - Physiological adaptations to fasting in an actively wintering canid, the Arctic blue fox (Alopex lagopus). AB - This study investigated the physiological adaptations to fasting using the farmed blue fox (Alopex lagopus) as a model for the endangered wild arctic fox. Sixteen blue foxes were fed throughout the winter and 32 blue foxes were fasted for 22 d in Nov-Dec 2002. Half of the fasted blue foxes were food-deprived again for 22 d in Jan-Feb 2003. The farmed blue fox lost weight at a slower rate (0.97-1.02% body mass d(-1)) than observed previously in the arctic fox, possibly due to its higher initial body fat content. The animals experienced occasional fasting induced hypoglycaemia, but their locomotor activity was not affected. The plasma triacylglycerol and glycerol concentrations were elevated during phase II of fasting indicating stimulated lipolysis, probably induced by the high growth hormone concentrations. The total cholesterol, HDL- and LDL-cholesterol, urea, uric acid and total protein levels and the urea:creatinine ratio decreased during fasting. Although the plasma levels of some essential amino acids increased, the blue foxes did not enter phase III of starvation characterized by stimulated proteolysis during either of the 22-d fasting procedures. Instead of excessive protein catabolism, it is liver dysfunction, indicated by the increased plasma bilirubin levels and alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase activities, that may limit the duration of fasting in the species. PMID- 16358269 TI - Effects of constant and fluctuating temperatures on egg survival and hatchling traits in the northern grass lizard (Takydromus septentrionalis, Lacertidae). AB - To understand how nest temperatures influence phenotypic traits of reptilian hatchlings, the effects of fluctuating temperature on hatchling traits must be known. Most investigations, however, have only considered the effects of constant temperatures. We incubated eggs of Takydromus septentrionalis (Lacertidae) at constant (24 degrees C, 27 degrees C, 30 degrees C and 33 degrees C) and fluctuating temperatures to determine the effects of these thermal regimes on incubation duration, hatching success and hatchling traits (morphology and locomotor performance). Hatching success at 24 degrees C and 27 degrees C was higher, and hatchlings derived from these two temperatures were larger and performed better than their counterparts from 30 degrees C and 33 degrees C. Eggs incubated at fluctuating temperatures exhibited surprisingly high hatching success and also produced large and well-performed hatchlings in spite of the extremely wide range of temperatures (11.6-36.2 degrees C) they experienced. This means that exposure of eggs to adversely low or high temperatures for short periods does not increase embryonic mortality. The variance of fluctuating temperatures affected hatchling morphology and locomotor performance more evidently than did the mean of the temperatures in this case. The head size and sprint speed of the hatchlings increased with increasing variances of fluctuating temperatures. These results suggest that thermal variances significantly affect embryonic development and phenotypic traits of hatchling reptiles and are therefore ecologically meaningful. PMID- 16358272 TI - Sperm-derived sperm motility-initiating substance from amphioxus Branchiostoma belcheri. AB - The sperm of amphioxus, Branchiostoma belcheri, were immotile when excised from the testis and suspended in seawater. The sperm became motile upon spawning in natural seawater, suggesting mechanisms that triggered sperm motility during spawning. When a male amphioxus that underwent spawning was transferred to a cup containing a small amount of natural seawater, and then the seawater containing the spawned sperm was centrifuged, the supernatant caused motility initiation in the immotile sperm from the testis. This sperm motility-initiating activity was also found in the supernatant of seawater in which immotile sperm from the testis were incubated overnight. These suggest that in the amphioxus, a sperm motility initiating substance resides in the sperm, and upon spawning, the substance is transformed into a free and active form to activate the sperm. Partial purification of the substance revealed it as a small and heat-stable substance with maximum UV absorbance at 234 nm. PMID- 16358270 TI - Viral transgenesis of embryonic cell cultures from the freshwater microcrustacean Daphnia. AB - The aim of this study was to determine whether a recombinant vesicular stomatitis virus (VSV) vector encoding a transgene could be used to infect and express a foreign gene in embryonic primary cell cultures derived from the freshwater microcrustacean Daphnia, the most widely used ecotoxicological model organism. To facilitate the evaluation of gene transfer, a reproducible method for establishing primary cultures from Daphnia embryonic tissues was developed. Within 24 hr after infection, transgene expression could be detected in cell culture. VSV was found to replicate in the cells with no apparent cytopathic effect. Here we report the first evidence of gene transfer and foreign gene expression in cultures of Daphnia embryonic cells using a recombinant viral vector. PMID- 16358273 TI - Embryonic gonadal and sexual organ development in a small viviparous skink, Niveoscincus ocellatus. AB - The majority of research into the timing of gonad differentiation (and sex determination) in reptiles has focused on oviparous species. This is largely because: (1) most reptiles are oviparous; (2) it is easier to manipulate embryonic developmental conditions (e.g., temperature) of eggs than oviductal embryos and (3) modes of sex determination in oviparous taxa were thought to be more diverse since viviparity and environmental sex determination (ESD)/temperature-dependent sex determination (TSD) were considered incompatible. However, recent evidence suggests the two may well be compatible biological attributes, opening potential new lines of enquiry into the evolution and maintenance of sex determination. Unfortunately, the baseline information on embryonic development in viviparous species is lacking and information on gonad differentiation and sexual organ development is almost non-existent. Here we present an embryonic morphological development table (10 stages), the sequence of gonad differentiation and sexual organ development for the viviparous spotted snow skink (Niveoscincus ocellatus). Gonad differentiation in this species is similar to other reptilian species. Initially, the gonads are indifferent and both male and female accessory ducts are present. During stage 2, in the middle third of development, differentiation begins as the inner medulla regresses and the cortex thickens signaling ovary development, while the opposite occurs in testis formation. At this point, the Mullerian (female reproductive) duct regresses in males until it is lost (stage 6), while females retain both ducts until after birth. In the later stages of testis development, interstitial tissue forms in the medulla corresponding to maximum development of the hemipenes in males and the corresponding regression in the females. PMID- 16358274 TI - GFP reporter gene confirms paternity in the androgenote Buenos Aires tetra, Hemigrammus caudovittatus. AB - A protocol for successful induction of androgenetic cloning of the Buenos Aires tetra (BT), Hemigrammus caudovittatus, with contrasting gray and golden strains is described. At the intensity of 4.2 W/m(2), UV irradiation for 2.75 min totally inactivated the maternal genome in eggs of gray BT. Following activation by sperm of golden BT, the 25-min-old embryos were shocked at 41 degrees C for 2 min to restore diploidy. Interestingly, the hatching success of the haploid fry was always higher than that of the diploid fry, indicating that the enhanced homozygosity (Y(2)Y(2)) is more deleterious than haploidy. Maternal genomic inactivation was confirmed by (i) expression of green fluorescent protein (GFP) gene in the 6-16 hr old live haploid and aneuploid embryos, (ii) golden body color in the diploid fry and adult and (iii) progeny testing. Survival of androgenotes was 10% at hatching and 6% at sexual maturity. Reproductive performance of F(0) and F(1) males (Y(2)Y(2)) was superior to that of normal ones (X(1)Y(2)), but that of the F(0) and F(1) females (X(2)X(2)) was inferior to the control (X(1)X(2)). Of 21 crosses involving homozygous androgenetic (Y(2)Y(2)) males and heterozygous (X(1)X(2)) females, 7 of them (33%) produced 3-9% unexpected female progenies. But only a single cross (14%) generated 3-4% unexpected female progenies, when 7 pairs of homozygous androgenetic (Y(2)Y(2)) males and (X(2)X(2)) females were crossed. Hence, the paternal autosomes, inherited by the homozygous androgenetic female (X(2)X(2)), produced female progenies in significantly less number of crosses, also at lower frequencies than the crosses with heterozygous females (X(1)X(2)), which carried an equal number of paternal and maternal autosomes. However, progenies resulting from the cross between gray female (X(1)X(2)) and golden male (Y(2)Y(2)), after undergoing androgenesis, were males, with paternal chromosomes alone, indicating that the presence of Y(2)Y(2) appears to override the modifying effect of autosomes, but the paternal or maternal autosomes seemed to override the single Y(2) present with X(1) or X(2), and induced the production of unexpected female progenies. Using Double sex Mab3 related transcription factor (DMRT 1)-specific primers, PCR analyses of the genomic DNA of the normal (X(1)Y(2)) and androgenetic males (X(1)Y(2)) produced two amplicons of 237 and 300 bp length. However, they were not detectable in the female (X(1)X(2)) genomic DNA, which amplified only one amplicon of 100 bp. Genomic DNA extracted from the 18 unexpected female progenies expressed the (X(1)Y(2)) genotype-specific banding pattern with two amplicons of 237 and 300 bp length and thereby confirmed that they were genotypic males. A partial sequencing of the male-specific sequence indicated that DMRT 1-specific primer was bound to the fragment of the genomic DNA of the male tetra, although the male-specific sequence of DMRT 1 was not completely detectable. PMID- 16358275 TI - Effect of human gonadotropins on spermiation and androgen biosynthesis in the testis of the toad Bufo arenarum (Amphibia, Anura). AB - This paper analyzes, in the toad Bufo arenarum, the effect on spermiation and androgen secretion of two human recombinant gonadotropins, human recombinant LH (hrLH) and human recombinant FSH (hrFSH) as well as the well-known spermiation inducing hormone, human chorionic gonadotropin (hCG). For this purpose, testes were incubated with different concentrations of hrLH (0.01-2.5 microg/ml) and hrFSH (0.05-5 microg/ml), and results were compared with those obtained with 2.5 microg/ml hCG. Spermiation was most efficiently stimulated by hrFSH, which elicited a higher response than either hrLH or hCG. Both hrFSH and hrLH produced a bell-shaped dose-response curve, with a 50% inhibition on spermiation at a concentration twice higher than that necessary to get the highest response. However, none of the gonadotropins yielded a biphasic response on androgen secretion, hrLH producing the highest response at a concentration that evoked a 70% inhibition in the spermiation test. Regarding steroidogenesis, hrLH and hrFSH were more active than hCG. Taken together, the results described in this paper suggest that, in B. arenarum, spermiation and androgen secretion are mediated by different receptors. After comparing the effects of recombinant hormones, we conclude that hrFSH has a greater effect on spermiation than hCG or hrLH. PMID- 16358276 TI - Chronic administration of leptin in Asian Blue Quail. AB - It is well known that leptin has the capacity to reduce food intake, cause body weight loss, and increase energy expenditure in several vertebrate species. In this study, we investigated the effects of chronically elevated leptin levels on behavior and physiology of Asian Blue Quail (Coturnix chinensis). Fifteen male quail were treated with chicken leptin dissolved in phosphate-buffered saline (PBS) via subcutaneously inserted osmotic pumps that released approximately 1 microg/g body weight/day during a 14-day period. Another 15 males acted as controls and their pumps released PBS only. All males were housed together with two females. We observed a decrease in body weight and feeding behavior in leptin treated birds, but not in control birds, after 2 days of treatment. Thereafter, all birds increased in weight. Males treated with leptin were more active and more likely to preen the day after the beginning of the treatment. Plasma cholesterol levels in leptin birds decreased during the first week of treatment and plasma triglycerides tended to remain lower compared to the controls during the whole 2-week period of treatment. Glucose levels appeared stable during the observation period. Leptin-treated males remained closer to accompanying females than did control males, and females together with leptin males took longer to lay their first egg compared to females together with control males. This is the first article showing the effect of leptin on cholesterol and triglyceride levels in birds. We also observed a change in the activity and male-female interaction pattern in tested quail. PMID- 16358277 TI - Effects of eyestalk ablation on carbonic anhydrase activity in the euryhaline blue crab Callinectes sapidus: neuroendocrine control of enzyme expression. AB - Carbonic anhydrase (CA) activity in the gills of the euryhaline blue crab, Callinectes sapidus, was measured in response to acute low-salinity transfer and treatment with eyestalk ablation (ESA) in an attempt to elucidate potential regulatory mechanisms of salinity-mediated CA induction. ESA alone resulted in an approximate doubling of CA activity in the posterior, ion-transporting gills of crabs acclimated to 35 ppt. Transfer of intact crabs to 28 ppt, a salinity at which the blue crab is still an osmotic and ionic conformer, had no effect on CA activity, but treatment with ESA prior to transfer resulted in a 5-fold increase. Hemolymph osmolality was unaffected by ESA. There was a 7-fold induction of CA activity in posterior gills of intact crabs transferred from 35 to 15 ppt, and this was potentiated by about 100% by ESA. Hemolymph osmolality was slightly elevated in the ESA-treated crabs. CA activity in anterior gills did not increase in response to any treatment. Hemolymph concentrations of methyl farnesoate (MF) were measured for all experimental animals. MF concentrations were undetectable in all intact crabs, regardless of salinity. Treatment with ESA resulted in elevated levels of hemolymph MF, but these levels were still relatively low and unrelated to salinity. These results suggest that CA induction is under the control of a regulatory substance located in the eyestalk. This substance appears to be a CA repressor, keeping CA expression at low levels in the gills of crabs acclimated to high salinity. Exposure to low salinity, or treatment with ESA, removes the effects of this putative repressor and allows CA induction to occur. PMID- 16358278 TI - Compensatory extension of gestation length with advance of conception in red deer (Cervus elaphus). AB - Calving date in many mammals is matched to the time of greatest food availability. Out of season calving results in heavy penalties in terms of own and offspring survival or body condition. This study examined whether gestation length is affected by advancing fertilisation. Thirty-six red deer hinds (of the Iberian and Scottish subspecies) were subjected to a synchronisation treatment of oestrus, ovulation, and artificial insemination on three dates, with remaining non-pregnant females mated with an intact male in a last group. Gestation was longer the more the fertilisation was advanced; gestation lasted 241.5+/-1.3 days (d) in the first group, 237.4+/-1.2 d in the second, 235.1+/-1.3 d in the third, and 231.2+/-1.6 d in the last. Mean gestation lasted 234.2+/-0.7 d. Hinds gained less weight during gestation the more the fertilisation was advanced. The difference was due at least in part to net body weight of the hind after calving compared to that at mating, and calves did not differ in birth weight. As early born calves suffer greater mortality in the field, this enlargening of gestation might be a compensatory response of the hinds to match calving with food availability. Under natural conditions, similar small modifications of gestation length may help hinds to overcome short-term adverse conditions for calving. Because calf mortality is correlated with birth weight, hinds may have kept calf birth weight constant at the expense of greater body weight loss. PMID- 16358279 TI - Two-step cell patterning on planar and complex curved surfaces by precision spraying of polymers. AB - Controlling adhesion of living animal cells plays a key role in biosensor fabrication, drug-testing technologies, basic biological research, and tissue engineering applications. Current techniques for cell patterning have two primary limitations: (1) they require photolithography, and (2) they are limited to patterning of planar surfaces. Here we demonstrate a simple, precision spraying method for both positive and negative patterning of planar and curved surfaces to achieve cell patterns rapidly and reproducibly. In this method, which we call precision spraying (PS), a polymer solution is aerosolized, focused with sheath airflow through an orifice, and deposited on the substrate using a deposition head to create approximately 25 microm sized features. In positive patterning, adhesive molecules, such as laminin or polyethylenimine (PEI) were patterned on polydimethylsiloxane (PDMS) substrates in a single spraying operation. A variety of animal cell types were found to adhere to the adhesive regions, and avoid the non-adhesive (bare PDMS) regions. In negative patterning, hydrophobic materials, such as polytetrafluoroethylene (PTFE) and PDMS, were patterned on glass substrates. Cells then formed patterns on the exposed glass regions and avoided the hydrophobic regions. Cellular patterns were maintained for up to 2 weeks in the presence of serum, which normally fouls non-adhesive regions. Additionally, we found that precision spraying enabled micropatterning of complex-curved surfaces. Our results show that precision spraying followed by cell plating enables rapid and flexible cellular micropatterning in two simple steps. PMID- 16358280 TI - A thermoregulatory center in hornets: IR photography. AB - In the Oriental hornet Vespa orientalis (Hymenoptera, Vespinae), there is on the dorsal side of the thorax, beneath the mesoscutum plate of the prothorax and around the median notal suture, a lump that, in the course of hornet activity, is warmer by 9 degrees C from the surrounding milieu and by up to 6 degrees C from other body parts of the hornet. This lump is about 1 mm in diameter, butterfly shaped, and its upper, posterior border abuts the base of the forewings. During hornet activity and via Infra Red photography one can observe heat extensions stemming from the center of the lump and proceeding forward in the direction of the head, downward toward the legs and backwards toward the bases of the wings. The warmest region is the center of the lump, with its margins showing a lower temperature. As for the legs of the hornet, their upper part is warmer than the other parts. The temperature gradients along the hornet's body are dependent on the extent and nature of hornet activity. Thus, during flight or ventilation activity, the thorax is the warmest part of the body, while the wings, legs, and antennae, as well as the posterior part of the gaster are colder, yet all these body parts are still warmer to varying degrees than the surrounding milieu. Thus, at night, when sentry worker hornets stand guard around the nest entrance and remain practically motionless, the temperature differences between the various body parts are retained unchanged. We conjecture that the described butterfly shaped lump is a thermoregulatory center (TC), which is neurogenically activated, since the changes occurring in it are rapid, a matter of one to several seconds and do not appear to be directly dependent on the hemolymph supply. The thermoregulatory center keeps a high constant temperature apparently related to hornet activity and the environmental conditions. The temperature cascade is most probably regulated via the tracheal system. Apparently another system activated by a heat pump mechanism keeps the gaster at a lower temperature than the environment. PMID- 16358282 TI - Inhibition of melanin synthesis pathway by tricyclazole increases susceptibility of Fonsecaea pedrosoi against mouse macrophages. AB - Fonsecaea pedrosoi produces melanin, a pigment related to virulence in pathogenic fungi. To understand the involvement of melanin in the protection of fungi, the authors used tricyclazole to inhibit the melanin pathway in F. pedrosoi. Experiments of pigmentation suggested that F. pedrosoi uniquely produces dihydroxynaphthalene-melanin. Pigments produced on cultures modified or not with tricyclazole were extracted by an alkali-acid method and submitted to infrared and ion exchange chromatography analysis; also cytochemistry analysis for cationized ferritin of whole cells was carried out. This group of experiments showed that the tricyclazole treatment on F. pedrosoi produced a melanin-like pigment, but less negatively charged and with less affinity for iron ions than that without the tricyclazole treatment, and this in turn lead to a less negatively charge cell wall surface. Scanning electron microscopy of such pigments showed that the melanin from control cultures maintained their hyphae like structures, which have been described as "melanin-ghosts," whereas the tricyclazole pigment showed an amorphous surface. Interaction of conidia from cultures of F. pedrosoi, modified by tricyclazole or not, with peritoneal activated macrophages suggested that tricyclazole causes higher association of fungus with macrophages, weakens the fungus capacity to destroy the macrophages, and diminishes the resistance to dry fracture procedures on samples prepared for high resolution scanning electron microscopy. PMID- 16358281 TI - Immunohistochemical study on the distribution of sodium-dependent vitamin C transporters in the respiratory system of adult rat. AB - As vitamin C (L-ascorbic acid, VC) is known to be essential for many enzymatic reactions, the study on the transport mechanism of VC through cytoplasmic membrane is crucial to understanding physiological role of VC in cells and the respiratory system. In this regard, the study on the newly identified sodium dependent VC transporters (SVCTs), SVCT1 and SVCT2, is required in organs that contain high concentration of VC. We have shown the distribution of SVCT proteins in the respiratory system, which has been reported to be one of the organs with a high concentration of VC, using immunohistochemical techniques. In the present study, intense SVCT immunoreactivities (IRs) were mainly localized in the respiratory system epithelial cells. In the trachea, both SVCT1 and 2 were localized in the psuedostratified ciliated columnar epithelium. In the terminal bronchiole, SVCT1 and 2 IRs were mainly observed in the apical portion of the simple columnar epithelium. In addition, SVCT IRs was localized within the cell membrane of some alveolar cells, even though we could not identify the exact cell types. These results provide the first evidence that intense SVCT1 and 2 IRs were found in the apical portion of the respiratory epithelial cells, suggesting that SVCT proteins in the apical portion could transport the reduced form of VC included in the airway surface liquid into the respiratory epithelial cells. PMID- 16358283 TI - Imaging TOF-SIMS of rat kidney prepared by high-pressure freezing. AB - Phosphocholine, potassium ions, and sodium ions were localized in rat kidney with imaging TOF-SIMS. Tissue preparation was performed with high-pressure freezing, freeze-fracturing and freeze-drying. The distribution of sodium ions was visualized by imaging the signal at m/z 23 of positively charged secondary ions, and the distribution of potassium ions was visualized by imaging the signal at m/z 39. Potassium was found localized within cells of the proximal tubulus epithelium and within cells of the glomeruli. High signals of sodium ions were seen in the interstitial tissue and also in epithelial cells of the collecting ducts and in glomeruli. The overlay image showed that the distribution of sodium ions and potassium ions were largely complementary with color mixing in glomeruli and in the interstitium surrounding proximal tubules. The ion distribution was further analyzed by correlation analysis. Phosphocholine-containing phospholipids were visualized by imaging the phosphocholine head group at m/z 184 of positively charged ions. The m/z 184 signal shows a ubiquitous distribution with a high intensity of phosphocholine in epithelial cells. Overlay image of m/z 184, m/z 39, and m/z 23 and multivariate analysis showed that the localization of high levels of phosphocholine colocalizes with high levels of potassium ions, as expected for an ion with intracellular localization. PMID- 16358284 TI - Study of rice pollen grains by multispectral imaging microscopy. AB - Cellular images obtained by light microscopy have been analyzed qualitatively, but there is still a lack of quantitative information about the variations in cellular metabolism of selective substances. A new approach using the multispectral imaging microscope (MIM) to observe rice (Oryza sativa L.) pollen grains is reported. A liquid crystal tunable filter device was used for wavelength selection from 400 to 720 nm and a cooled two-dimensional monochrome charge coupled device for image detection. Rice pollen were stained respectively by acetocarmine, Coomassie blue, or iodine potassium-iodine, and then imaged by MIM. The images were processed by the WuDa Image Analysis System 2003 (computer software), and the transmittance spectra for pollen grain images were obtained. The statistical analysis of the transmittance data showed that the macromolecular amount (nucleic acid, protein, starch) of male-sterile line (MSL) pollen grains was less than those of the fertility-maintaining line (FML). For instance there was a significantly lower nucleic acid content in the MSL than in the FML pollen. The results revealed that pollen abortion was directly related to the diminution of intracellular substances for metabolism. Consequently, we have established a quantitative criterion to determine pollen sterility. Comparing the spectra features of the FML with the MSL, we found that certain spectra features can be used to identify various types of abortion pollens and the deficient cytoplasm of male-sterile rice. Our experimental results offer the first quantitative understanding for evaluating cell morphological structure correlated with cellular physiological status. PMID- 16358285 TI - The destination of the aged, nonreleasable neurohypophyseal peptides stored in the neural lobe is associated to the remodeling of the neurosecretory axon. AB - The present investigation was designed to investigate the fate of the large pool of neurohypophyseal hormones that is never released into the blood. Normal Sprague-Dawley and taiep mutant rats were investigated under normal water balance, after dehydration and after dehydration-rehydration. Lectin histochemistry and light- and electron-microscopic immunocytochemistry using antibodies against vasopressin, oxytocin, and neurophysins used at low (1:1,000) and high (1:15,000) dilutions allowed to distinguish (1) recently packed immature granules, as those located in the perikaryon; (2) mature; and (3) aged granules. The distribution of these granules within the different domains of the neurosecretory axons located in the neural lobe, namely, undilated segments, swellings, terminals, and Herring bodies, and the response of these compartments to dehydration and dehydration-rehydration allowed to roughly follow the routing of the granules through such axonal domains. It is suggested that granules may move backward and forward between the terminals and the swellings. At variance, aged granules located in Herring body are retained in this compartment and would finally become degraded. Herring bodies displayed distinct lectin binding and immunocytochemical properties, allowing to distinguish them from axonal swellings. After a dehydration-rehydration cycle, immunocytochemistry and electron microscopy revealed that Herring bodies were no longer present in the neural lobe and that several terminals had degenerated. It is concluded that (1) the neurophysin axons may undergo remodeling under appropriate stimuli and (2) Herring bodies are a specialized and plastic domain of the magnocellular neurosecretory neuron involved in the disposal of aged neurosecretory granules. No differences were detected at the neural lobe level between normal and mutant rats subjected to the same experimental conditions. PMID- 16358286 TI - Chloroplast ultrastructure in leaves of Urtica dioica L. analyzed after high pressure freezing and freeze-substitution and compared with conventional fixation followed by room temperature dehydration. AB - In this article, we report on the adaptation of high-pressure freezing and freeze substitution (HPF-FS) for ultrastructural analysis of leaf tissue with special emphasis on chloroplasts. To replace the gas in the intercellular spaces, a mixture of water and methanol (MeOH) was employed. We compared three different supplements for FS--osmiumtetroxide, uranyl acetate, and safranin--with regard to the preservation of the ultrastructure of chloroplasts and other cellular compartments. The results show that (i) replacement of air within intercellular spaces by 8% (v/v) MeOH has no influence on the ultrastructure of the chloroplasts, (ii) undulation of membranes frequently observed after conventional preparation of specimens does not occur during chemical fixation but during room temperature dehydration, and (iii) uranyl acetate or osmium tetroxide employed during FS are not superior over safranin. PMID- 16358287 TI - Optimization of the cell seeding density and modeling of cell growth and metabolism using the modified Gompertz model for microencapsulated animal cell culture. AB - Cell microencapsulation is one of the promising strategies for the in vitro production of proteins or in vivo delivery of therapeutic products. In order to design and fabricate the optimized microencapsulated cell system, the Gompertz model was applied and modified to describe the growth and metabolism of microencapsulated cell, including substrate consumption and product formation. The Gompertz model successfully described the cell growth kinetics and the modified Gompertz models fitted the substrate consumption and product formation well. It was demonstrated that the optimal initial cell seeding density was about 4-5 x 10(6) cells/mL of microcapsule, in terms of the maximum specific growth rate, the glucose consumption potential and the product formation potential calculated by the Gompertz and modified Gompertz models. Modeling of cell growth and metabolism in microcapsules provides a guideline for optimizing the culture of microencapsulated cells. PMID- 16358288 TI - Oxygen supply for CHO cells immobilized on a packed-bed of Fibra-Cel disks. AB - Packed-bed bioreactors (PBR) have proven to be efficient systems to culture mammalian cells at very high cell density in perfusion mode, thus leading to very high volumetric productivity. However, the immobilized cells must be continuously supplied with all nutrients in sufficient quantities to remain viable and productive over the full duration of the perfusion culture. Among all nutrients, oxygen is the most critical since it is present at very low concentration due to its low solubility in cell culture medium. This work presents the development of a model for oxygenation in a packed-bed bioreactor system. The experimental system used to develop the model was a packed-bed of Fibra-Cel disk carriers used to cultivate Chinese Hamster Ovary cells at high density ( approximately 6.1 x 10(7) cell/mL) in perfusion mode. With the help of this model, it was possible to identify if a PBR system is operated in optimal or sub-optimal conditions. Using the model, two options were proposed, which could improve the performance of the basal system by about twofold, that is, by increasing the density of immobilized cells per carrier volume from 6.1 x 10(7) to 1.2 x 10(8) cell/mL, or by increasing the packed-bed height from 0.2 to 0.4 m. Both strategies would be rather simple to test and implement in the packed-bed bioreactor system used for this study. As a result, it would be possible to achieve a substantial improvement of about twofold higher productivity as compared with the basal conditions. PMID- 16358289 TI - 1H and 13C NMR signal assignments of a novel Baeyer-Villiger originated diterpene lactone. AB - A highly rearranged novel dilactone was the single product isolated from Baeyer Villiger oxidation of a norketone prepared from grandiflorenic acid, a natural kaurane diterpene. The complete 1H and 13C NMR assignment is presented for this novel compound that showed discrete in vitro antibacterial activity. PMID- 16358290 TI - Real-life applications of the MULVADO software package for processing DOSY NMR data. AB - MULVADO is a newly developed software package for DOSY NMR data processing, based on multivariate curve resolution (MCR), one of the principal multivariate methods for processing DOSY data. This paper will evaluate this software package by using real-life data of materials used in the printing industry: two data sets from the same ink sample but of different quality. Also a sample of an organic photoconductor and a toner sample are analysed. Compared with the routine DOSY output from monoexponential fitting, one of the single channel algorithms in the commercial Bruker software, MULVADO provides several advantages. The key advantage of MCR is that it overcomes the fluctuation problem (non-consistent diffusion coefficient of the same component). The combination of non-linear regression (NLR) and MCR can yield more accurate resolution of a complex mixture. In addition, the data pre-processing techniques in MULVADO minimise the negative effects of experimental artefacts on the results of the data. In this paper, the challenges for analysing polymer samples and other more complex samples will also be discussed. PMID- 16358291 TI - Discrimination of enantiomers by means of NMR spectroscopy using chiral liquid crystalline solution: application to triazole fungicides, uniconazole and diniconazole. AB - An NMR method for discriminating among enantiomers by using a chiral liquid crystalline solution was applied to chiral triazole compounds, uniconazole (1) and diniconazole (2), which exhibit antifungal and plant growth regulating activities. These chiral compounds were dissolved in PBLG (poly-gamma-benzyl-L glutamate)--CDCl3 chiral liquid crystalline solvent for measurements of 13C NMR. The enantiomeric separations were primarily observed in the signals of aromatic carbons owing to differences in chemical shift anisotropies. The enantiomeric excess (ee) was determined from the integral scale of the separated peaks. The resulting ee values are in fair agreement with the actual values. The extrasplittings due to residual dipolar couplings were also measured using Het2DJ spectra for 1S and R, and 2R, and the results are discussed. PMID- 16358292 TI - Complete NMR signal assignments of flavonol derivatives. AB - Common substitution positions of flavonols are at C-5 and C-7; 6-substituted flavonol derivatives are rarely found in natural sources. Here, we report complete assignments of 1H and 13C chemical shifts of eight flavonol derivatives including four 6-substituted flavonols. PMID- 16358294 TI - Monosaccharides as internal probes for the determination of the absolute configuration of 2-butanol. AB - D-Glucose, D-mannose and L-rhamnose were reacted with a racemic mixture of 2 butanol, and the resulting alpha-glycosides were analyzed by 1H NMR with COSY and NOESY experiments. Conformational analysis of alpha-glycosidic bonds performed with molecular modeling and appropriate heteronuclear long-range coupling measurements and combined with analysis of dipolar couplings observed in NOESY spectra allowed the assignment of absolute configuration in the aglycones of elucidated alpha-glycosides. PMID- 16358293 TI - Structural determination of 3beta-stearyloxy-urs-12-ene from Maytenus salicifolia by 1D and 2D NMR and quantitative 13C NMR spectroscopy. AB - Six pentacyclic triterpenoids, 3beta-stearyloxy-urs-12-ene (1), friedelin (2), 3beta-friedelinol (3), alpha-amyrin (4), beta-amyrin (5), and lupeol (6), have been isolated from the hexane extract of Maytenus salicifolia Reissek (Celastraceae) leaves. The molecular and structural formula as well as the stereochemistry of a new pentacyclic triterpene (1) were determined using data obtained from 1H and 13C NMR spectra, DEPT135 and by 2D HSQC, HMBC, COSY and NOESY experiments. The molecular formula C48H84O2 was established using quantitative 13C NMR, and the molecular weight (692 Da) was confirmed by elemental analysis and mass spectrometry (GC-MS). PMID- 16358295 TI - Towards the automatic analysis of NMR spectra: part 6. Confirmation of chemical structure employing both 1H and 13C NMR spectra. AB - A method of comparing predicted and experimental chemical shifts was used to confirm or refute postulated structures. 1H NMR spectra returned all true positives with a false positive rate of 4%. When an analogous procedure was adopted for 13C NMR spectra, the false positive rate dropped to 1%, whereas the more practical HSQC data yielded a false positive rate of 2%. If the HSQC results were combined with 1H results, a false positive rate of 1% resulted, 4 times more accurate than 1H alone. PMID- 16358296 TI - Diastereoisomerism of thiol complexes of arsenic acids and pseudoasymmetry of arsenic: a 1H and 13C NMR study. AB - Diastereoisomeric complexes of methylphenylarsinic acid and (L)-glutathione could be partially separated by HPLC, but the separated compounds rapidly racemized, presumably by pyramidal inversion at the arsenic atom. Hydrolysis of the diastereoisomeric complexes yielded methylphenylarsinous acid as a pair of enantiomers revealed by a 1H NMR study with an asymmetric lanthanide shift reagent. Methylphenylarsinous acid was also synthesized as an enantiomeric pair, shown by an asymmetric shift reagent experiment, by the hydrolysis of iodomethylphenylarsine. 1H and 13C NMR spectroscopy were used to demonstrate that complexing of phenylarsonic acid with (R,S)-3-mercapto-1,2-propanediol and with (R,S)-1-mercapto-2-propanol yielded, in each case, a pair of enantiomers, PhAs[(R)-ligand)]2, PhAs[(S)-ligand)]2, in which the homomorphic ligands were diastereotopic, and a pair of diastereoisomers, PhAs[(R)-ligand][(S)-ligand], which differed from each other in the configuration about the pseudoasymmetric arsenic atom. PMID- 16358297 TI - Intensity-modulated radiation therapy in head and neck cancers: an update. AB - Intensity-modulated radiation therapy (IMRT), an advent of three-dimensional conformal radiotherapy (3D CRT), has excited the profession of radiation oncology more than any other new invention since the introduction of the linear accelerator. Approximately 1000 articles have been published on this topic to date, more than 200 of which focus on head and neck cancer. IMRT is based on computer-optimized treatment planning and a computer-controlled treatment delivery system. The computer-driven technology generates dose distributions that sharply conform to the tumor target while minimizing the dose delivered to the surrounding normal tissues. The high dose volume that tailors to the 3D configuration of the tumor along with the ability to spare the nearby normal tissues allows the option of tumor dose escalation. The head and neck region is an ideal target for this new technology for several reasons. First, IMRT offers the potential for improved tumor control through delivery of high doses to the target volume. Second, because of sharp dose gradients, IMRT results in the relative sparing of normal structures, such as the parotid glands, in the head and neck region. Third, organ motion is virtually absent in the head and neck region so, with proper immobilization, treatment can be accurately delivered. Although this is a relatively new technology, single-institution retrospective studies show better dosimetric profiles compared with conventional radiation techniques, as well as excellent clinical results. Salivary gland sparing using IMRT has also resulted in reduced incidence and severity of xerostomia, and this has been tested in a randomized trial against conventional radiotherapy for early stage nasopharyngeal cancer. The results do confirm that IMRT does decrease xerostomia compared with conventional radiotherapy. PMID- 16358299 TI - Biochemical and genetic characterization of Hmi1p, a yeast DNA helicase involved in the maintenance of mitochondrial DNA. AB - The HMI1 gene encodes a DNA helicase that localizes to the mitochondria and is required for maintenance of the mitochondrial DNA (mtDNA) genome of Saccharomyces cerevisiae. Identified based on its homology with E. coli uvrD, the HMI1 gene product, Hmi1p, has been presumed to be involved in the replication of the 80 kb linear S. cerevisiae mtDNA genome. Here we report the purification of Hmi1p to apparent homogeneity and provide a characterization of the helicase reaction and the ATPase reaction with regard to NTP preference, divalent cation preference and the stimulatory effects of different nucleic acids on Hmi1p-catalysed ATPase activity. Genetic complementation assays indicate that mitochondrial localization of Hmi1p is essential for its role in mtDNA metabolism. The helicase activity, however, is not essential. Point mutants that lack ATPase/helicase activity partially complement a strain lacking Hmi1p. We suggest several possible roles for Hmi1p in mtDNA metabolism. PMID- 16358300 TI - Ewing sarcoma bone metastases: 'you see one, you've seen them all': (commentary on Furth et al., page 607). PMID- 16358301 TI - Low efficacy of methotrexate in childhood acute myeloid leukemia (AML): single agent therapeutic window study in relapsed AML. AB - BACKGROUND: The efficacy in pediatric acute myeloid leukemia (AML) of single agent methotrexate (MTX) at a higher dose than previously applied, 1,000 mg/m2, given as a theoretically beneficial 36-hr continuous infusion, is unknown, but may be beneficial based on preclinical data. PROCEDURE: We performed a therapeutic window study in children with first relapsed AML treated in four different countries. RESULTS: Based on a comparison between the percentage of leukemic blasts in the bone marrow shortly before and 7-10 days after the MTX infusion, none of the first cohort of nine patients showed a good response, defined as a reduction of blasts of at least 50%. Therefore, the study was closed, concluding that the probability of a good response in this patient-group was most likely to be less than 30%. By that time, another four patients had been enrolled, of which one patient with a late relapsed AML FAB type M7 showed a good response. Toxicity of MTX was limited and tolerable. CONCLUSIONS: This study shows that single-agent MTX in the applied regimen in pediatric relapsed AML has limited efficacy. However, it also demonstrates the feasibility of an international and therapeutic window phase II study in pediatric relapsed AML. PMID- 16358302 TI - Incidence of catheter-related infections within 30 days from insertion of Hickman Broviac catheters. AB - AIM: To evaluate the incidence of surgical site infections and bacteremias occurring within 30 days from insertion of partially implanted central venous catheters. PATIENTS AND METHODS: Four hundred eighteen devices positioned in children with cancer or undergoing bone marrow transplant were followed prospectively. RESULTS: During a follow-up of 12,394 catheter-days, a total of 13 infectious episodes were documented, with an overall incidence of 3.1% and 1.05 episodes/1,000 catheter-days. Coagulase-negative staphylococci represented the causative pathogens of all episodes. Overall, surgical wound infections occurred in 1.4% of all catheters, with a rate of 0.48/1,000 catheter-days, while isolated bacteremias were observed in 1.7% of all inserted devices, with a rate of 0.57/1,000 catheter-days. CONCLUSIONS: Infections are rare events within 30 days from insertion of partially implanted central venous catheters and coagulase negative staphylococci represent the most frequently isolated cause of these complications. PMID- 16358303 TI - Overt testicular disease (OTD) at diagnosis is not associated with a poor prognosis in childhood acute lymphoblastic leukemia: results of the EORTC CLG Study 58881. AB - To assess the prognosis of overt testicular disease (OTD) at diagnosis of acute lymphoblastic leukemia (ALL), we analyzed the outcome of 1,165 boys enrolled in EORTC trial 58881. Thirteen (1.1%) boys had OTD associated with bad prognostic features. Patients with and without OTD did not differ in event-free survival (EFS) (P=0.30) or overall survival (OS) (P=0.54), even after adjustment for the three most important independent factors (NCI risk group, presence of very high risk features, type of asparaginase used). OTD was not an independent prognostic factor. These results may be due to the use of risk-adjusted intensive chemotherapy comprising high-dose methotrexate. PMID- 16358304 TI - 17O NMR spectra of some organotin(IV) compounds containing O,C,O-chelating ligands. AB - The 17O chemical shifts of substituted benzyl ethers and a set of organotin(IV) derivatives containing O,C,O-chelating ligands were studied. Measured 17O chemical shifts were correlated with the additivity substituent increments for carbon atoms in the alkyl groups, and intramolecular Sn-O distance was obtained by X-ray diffraction techniques in the solid state. PMID- 16358305 TI - 13C/15N distance determination by CPMAS NMR in uniformly 13C labeled molecules. AB - The REDOR and CPMAS techniques are applied for measuring 13C-15N dipolar coupling constants in glycine. It is shown that the selective CP or SPECIFIC CP technique removes the coherent evolution of the spin system under homonuclear 13C-13C J couplings. While the large coupling constant (approximately 900 Hz) is readily determined because of the presence of large oscillations in the CPMAS dynamics, their absence precludes the measurement of the small coupling constant (approximately 200 Hz). The experimental results and numerical simulations demonstrate that the determination of 13C-15N coupling constants of medium size (<1 kHz) by the CPMAS technique is mainly limited by the strength of the 1H decoupling field and the size of the 13C and 15N chemical shift anisotropies. PMID- 16358306 TI - 1H, 13C, and 73Ge NMR spectral analysis of substituted aryltrimethylgermanes. AB - NMR chemical shifts of 1H, 13C, and 73Ge are reported for a series of monosubstituted aromatic trimethylgermanes of the type XC6H4Ge(CH3)3; X = p N(CH3)2, p-OCH3, p-OC2H5, p-C(CH3)3, p-Si(CH3)3, p-Ge(CH3)3, p-Sn(CH3)3, p-CH3, m CH3, -H, m-OCH3, p-Cl, p-Br, m-F, m-CF3, p-CF3, o-OCH3, and o-CH3. The relatively narrow 73Ge resonances show a strong correlation with Hammett sigma constants, with a correlation coefficient of 0.976 and 0.876 for 73Ge chemical shifts in meta- and para-substituted derivatives, respectively. The 13C chemical shifts of the methyl carbons bonded to germanium also display a relationship, with correlation coefficients of 0.904, 0.993, and 0.911 for para-, meta- and all derivatives, respectively. Comparisons of the Hammett plots for the homologous series XC6H4M(CH3)3; M = C, Si, Ge, Sn, show that, in general, correlation coefficients decrease while slopes increase significantly down the group, presumably reflecting the corresponding increase in chemical shift range of the group 14 atom. The Hammett constant derived for the p-Ge(CH3)3 group of +0.13 compares with the NMR-derived constants of -0.12 for p-C(CH3)3, +0.14 for p Si(CH3)3, and -0.14 for p-Sn(CH3)3. The indication of electron release by carbon and tin can be rationalized through traditional hyperconjugative arguments for carbon and by the low electronegativity and consequent inductive effect of tin. The small electron attraction suggested by the positive constants for silicon and germanium can be simply, and perhaps naively, attributed to pi-acceptor interactions with the benzene ring. PMID- 16358307 TI - Utilisation of the budding yeast Saccharomyces cerevisiae for the generation and isolation of non-lethal ricin A chain variants. AB - Knowledge of the uptake, membrane translocation, refolding and ribosome interaction of the ribosome-inactivating toxin ricin is incomplete at the present time. Ricin A chain (RTA) is the catalytic subunit of holotoxin and is also of particular interest as a vaccine candidate. For many studies into the uptake and immunological applications of ricin, it is essential to have inactive variants. Here, following error-prone polymerase chain reaction of the RTA open reading frame, we have used a modified gap-repair protocol in Saccharomyces cerevisiae to show that it is possible to rapidly generate a panel of inactive RTA mutants. Since yeast cells have ribosomes that are highly sensitive to RTA, we utilized a genetic selection based on the viability of transformants. This enabled the recovery of a number of mutations, some not previously identified, which permitted production of full-length but non-toxic RTA proteins. Such disarmed toxins may have utility as tools to study the cytosolic entry and action of RTA, and as potential vaccine candidates. PMID- 16358308 TI - Rapid asymmetrical evolution of Saccharomyces cerevisiae wine yeasts. AB - Genetic instability causes very rapid asymmetrical loss of heterozygosity (LOH) at the cyh2 locus and loss of killer K2 phenotype in some wine yeasts under the usual laboratory propagation conditions or after long freeze-storage. The direction of this asymmetrical evolution in heterozygous cyh2(R)/CYH2(S) hybrids is determined by the mechanism of asymmetrical LOH. However, the speed of the process is affected by the differences in cell viability between the new homozygous yeasts and the original heterozygous hybrid cells. The concomitant loss of ScV-M2 virus in the LOH process may increase cell viability of cyh2(R)/cyh2(R) yeasts and so favour asymmetrical evolution. The presence of active killer K2 toxin, however, abolishes the asymmetrical evolution of the hybrid populations. This phenomenon may cause important sudden phenotype changes in industrial and pathogenic yeasts. PMID- 16358309 TI - Sensorineural hearing loss in a case of familial hemophagocytic lymphohistiocytosis. AB - Severe sensorineural hearing loss (bilateral >80 dB) was diagnosed in a case of familial hemophagocytic lymphohistiocytosis (FHL). The female patient developed HLH at 3 months of age and underwent allogeneic cord blood transplantation at 11 months of age following 7 months of immuno-chemotherapy. The type 2 FHL patient had a homozygous perforin gene mutation of 1090-1091delCT, and was noted to have hearing loss at 3.5 years of age. Retrospective evaluation did not clarify the exact causes of hearing loss. Reports on Kawasaki disease, suggesting a correlation between severe inflammatory status in infancy and the development of sensorineural hearing loss, may shed some light on this rare complication in this case of FHL. Considering the markedly improved prognosis of FHL due to recent advances made in the molecular diagnosis and in the management including allogeneic hematopoietic stem cell transplantation, auditor by screening might be warranted for surviving FHL patients. PMID- 16358310 TI - A proposal for an international retinoblastoma staging system. AB - BACKGROUND: Although intra-retinal tumor has long been staged presurgically according to the Reese-Ellsworth (R-E) system, retinoblastoma differs from other pediatric neoplasms in never having had a widely accepted classification system that encompasses the entire spectrum of the disease. Comparisons among studies that consider disease extension, risk factors for extra-ocular relapse, and response to therapy require a universally accepted staging system for extra ocular disease. PROCEDURE: A committee of retinoblastoma experts from large centers worldwide has developed a consensus classification that can encompass all retinoblastoma cases and is presented herein. Patients are classified according to extent of disease and the presence of overt extra-ocular extension. In addition, a proposal for substaging considering histopathological features of enucleated specimens is presented to further discriminate between Stage I and II patients. RESULTS: The following is a summary of the classification system developed-Stage 0: Patients treated conservatively (subject to presurgical ophthalmologic classifications); Stage I: Eye enucleated, completely resected histologically; Stage II: Eye enucleated, microscopic residual tumor; Stage III: Regional extension [(a) overt orbital disease, (b) preauricular or cervical lymph node extension]; Stage IV: Metastatic disease [(a) hematogenous metastasis: (1) single lesion, (2) multiple lesions; (b) CNS extension: (1) prechiasmatic lesion, (2) CNS mass, (3) leptomeningeal disease]. A proposal is also presented for substaging of enucleated Stages I and II eyes. CONCLUSIONS: The proposed staging system is the product of an international effort to adopt a uniform staging system for patients with retinoblastoma to cover the whole spectrum of the disease. PMID- 16358311 TI - Gene expression profiling reveals intrinsic differences between T-cell acute lymphoblastic leukemia and T-cell lymphoblastic lymphoma. AB - BACKGROUND: T-cell acute lymphoblastic leukemia (T-ALL) and T-cell lymphoblastic lymphoma (T-LL) and are often thought to represent a spectrum of a single disease. The malignant cells in T-ALL and T-LL are morphologically indistinguishable, and they share the expression of common cell surface antigens and cytogenetic characteristics. However, despite these similarities, differences in the clinical behavior of T-ALL and T-LL are observed. PROCEDURE: We analyzed the gene expression profiles of T-ALL and T-LL samples obtained from Children's Oncology Group (COG) tumor banks using DNA arrays. Immunohistochemistry was also performed to validate the expression of selected targets. RESULTS: Unsupervised hierarchical clustering of all samples showed complete segregation of T-ALL and T LL into distinct clusters. Next, we identified the top 201 genes that best differentiated T-ALL from T-LL using significance analysis of microarrays (SAM), a supervised statistical approach. Genes representing several functional groups were differentially expressed in T-LL and T-ALL. Prediction analysis of microarrays (PAM) identified a subset of genes, which accurately classified all 19 T-ALL and T-LL samples with an overall misclassification error rate of 0. Immunohistochemical validation of protein expression of selected genes identified by microarray analysis confirmed overexpression of MLL-1 in T-LL tumor cells compared to T-ALL and CD47 in T-ALL tumors cells when compared to T-LL. CONCLUSIONS: Despite significant similarities between the malignant T-cell precursors, clear differences in the gene expression profiles were observed between T-ALL and T-LL implying underlying differences in the biology of the two entities. PMID- 16358312 TI - Ventral hippocampal muscimol disrupts context-specific fear memory retrieval after extinction in rats. AB - Research aimed at understanding Pavlovian fear memory extinction has yielded considerable insight into the conditions under which fear memories may become inhibited. After extinction, Pavlovian fear memory retrieval is context-specific. Fear memories are not expressed in the extinction context, but they are expressed in every other context. Research indicates that the dorsal hippocampus mediates the context-specific expression of fear memory, but the role of the ventral hippocampus in mediating this process is unknown. Insofar as the ventral hippocampus is involved in the acquisition and expression of both context and tone fear, we asked whether GABA systems in the ventral hippocampus mediate context-specific fear memory retrieval after extinction. Experiment 1 showed that ventral hippocampal inactivation with muscimol disrupted context-specific fear memory retrieval. Experiment 2 showed that rats infused with muscimol can discriminate a context in which they were shocked from a neutral context. Nonetheless, they do appear to have a mild impairment in this task. Experiment 3 showed that ventral hippocampal muscimol did not disrupt locomotor activity, but did result in a slight increase in freezing and grooming, an effect that cannot explain the context-specific retrieval deficit demonstrated in experiment 1. These data are consistent with a role for the ventral hippocampus in mediating context-specific fear memory retrieval. PMID- 16358313 TI - Change in the shape and density of dendritic spines caused by overexpression of acidic calponin in cultured hippocampal neurons. AB - Dendritic spines are morphing structures believed to provide a cellular substrate for synaptic plasticity. It has been suggested that the actin cytoskeleton is the target of molecular mechanisms regulating spine morphology. Here we hypothesized that acidic calponin, an actin-binding protein, is one of the key regulators of actin filaments during spine plasticity. Our data showed that the overexpression of acidic calponin-GFP (green fluorescent protein) in primary cultures of rat hippocampal neurons causes an elongation of spines and an increase of their density as compared with those of GFP-expressing neurons. These effects required the actin-binding domains of acidic calponin. The close apposition of the presynatic marker synaptophysin to these long spines and the presence of specific postsynaptic markers actin, PSD-95, NR1, and GluR1 suggested the existence of functional excitatory synaptic contacts. Indeed, electrophysiological data showed that the postsynaptic overexpression of acidic calponin enhanced the frequency of miniature excitatory postsynaptic currents as compared with that of GFP expressing neurons, but did not affect their properties such as amplitude, rise time, and half width. Studies in heterologous cells revealed that acidic calponin reorganized the actin filaments and stabilized them. Taken together, these findings show that acidic calponin regulates dendritic spine morphology and density, likely via regulation of the actin cytoskeleton reorganization and dynamic. Furthermore, the acidic calponin-induced spines are able to establish functional glutamatergic synapses. Such data suggest that acidic calponin is a key factor in the regulation of spine plasticity and synaptic activity. PMID- 16358314 TI - Construction and characterization of a series of vectors for Schizosaccharomyces pombe. AB - A set of vectors was created to allow cloning and expression studies in Schizosaccharomyces pombe. These vectors had a uniform backbone with an efficient Sz. pombe ARS, ARS3002, but different selectable markers--his3+, leu1+, ade6+ and ura4+. The vectors functioned efficiently as autonomously replicating plasmids that could also be converted into integrating vectors. The ura4+-containing vector was used to construct a Sz. pombe genomic library. PMID- 16358315 TI - Effects of hippocampal lesions on the monkey's ability to learn large sets of object-place associations. AB - Earlier studies found that recognition memory for object-place associations was impaired in patients with relatively selective hippocampal damage (Vargha-Khadem et al., Science 1997; 277:376-380), but was unaffected after selective hippocampal lesions in monkeys (Malkova and Mishkin, J Neurosci 2003; 23:1956 1965). A potentially important methodological difference between the two studies is that the patients were required to remember a set of 20 object-place associations for several minutes, whereas the monkeys had to remember only two such associations at a time, and only for a few seconds. To approximate more closely the task given to the patients, we trained monkeys on several successive sets of 10 object-place pairs each, with each set requiring learning across days. Despite the increased associative memory demands, monkeys given hippocampal lesions were unimpaired relative to their unoperated controls, suggesting that differences other than set size and memory duration underlie the different outcomes in the human and animal studies. PMID- 16358316 TI - Impairment and recovery on a food foraging task following unilateral vestibular deafferentation in rats. AB - It has been suggested that the vestibular system may contribute to the development of higher cognitive function, especially spatial learning and memory that uses idiothetic cues (e.g., dead reckoning). However, few studies have been done using behavioral tasks that could potentially separate the animals' ability for dead reckoning from piloting. The food foraging task requires the animal to continuously monitor and integrate self-movement cues and generate an accurate return path. It has been shown that bilateral vestibular-lesioned rats were impaired on this task. The present study used the same task to further examine the contribution of vestibular information to spatial navigation by comparing unilateral and bilateral lesions and by testing the animals at different time points following the lesion. The results demonstrated that animals with unilateral vestibular deafferentation were impaired in performing the task in the dark at 3 months after the lesion, and this impairment disappeared at 6 months after the lesion. This supports the notion that vestibular information contributes to dead reckoning and suggests possible recovery of function over time after the lesion. Animals with bilateral vestibular deafferentation were not able to be tested on the foraging task because they exhibited behavior distinct from the unilateral-lesioned animals, with significant hesitation in leaving their home cage for as long as 6 months after the lesion. PMID- 16358318 TI - High serum values of soluble CD154, IL-2 receptor, RANKL and osteoprotegerin in Langerhans cell histiocytosis. AB - BACKGROUND: To determine useful biochemical markers in Langerhans cell histiocytosis (LCH), we analyzed the serum levels of soluble CD154 (sCD154), IL2 receptor (sIL2-R), receptor activator of NF-kappaB ligand (sRANKL), and osteoprotegerin (OPG). PROCEDURE: Our study included 46 newly diagnosed LCH patients (single-system multi-site (SM type): n = 20, and multi-system multi-site (MM type): n = 26) who were treated with the JLSG-02 protocol between 2002 and 2004. The median age of the patients was 3.8 years old (range 0-18). sCD154, sIL2 R, sRANKL, and OPG were measured by ELISA at diagnosis (n = 46) and after 6-weeks of induction therapy (n = 14). RESULTS: The values of sCD154, sIL-2R, sRANKL, and OPG, and the sRANKL/OPG ratio in sera were significantly higher in patients with LCH compared with controls (1.83 +/- 1.38 vs. 0.22 +/- 0.16 ng/ml, P < 0.001; 1,600 +/- 1,060 vs. 420 +/- 160 pg/ml, P < 0.001; 1.72 +/- 1.20 vs. 1.04 +/- 1.09 pmol/L, P = 0.019; 6.34 +/- 2.94 vs. 3.71 +/- 2.03 pmol/L, P < 0.001; and 0.40 +/ 0.45 vs. 0.16 +/- 0.17, P = 0.023, respectively). Serum levels of sIL-2R were significantly elevated in the MM type compared with the SM type (2,050 +/- 1,060 vs. 870 +/- 340 pg/ml, P < 0.001). Serum OPG levels were also significantly elevated in the MM type compared with the SM type (7.58 +/- 2.72 vs. 5.13 +/- 2.69 pmol/L, P = 0.008) and negatively correlated with the number of bone lesions (r = -0.56, P = 0.007). In contrast, the sRANKL/OPG ratios were significantly higher in the SM type than the MM type (0.57 +/- 0.54 vs. 0.19 +/- 0.14, P = 0.002) and positively correlated with the number of bone lesions (r = 0.34, P = 0.040). In patients who responded to the induction therapy, serum levels of sIL 2R, sRANKL, and OPG, and the sRANKL/OPG ratio decreased significantly after the therapy (1,170 +/- 600 vs. 730 +/- 290 pg/ml, P = 0.029; 2.19 +/- 1.06 vs. 1.24 +/- 0.66 pmol/L, P < 0.001; 6.13 +/- 2.40 vs. 4.72 +/- 2.03 pmol/L, P = 0.040; and 0.57 +/- 0.52 vs. 0.41 +/- 0.37, P = 0.02, respectively). In the three patients who did not respond to the induction therapy, the serum levels of sCD154 increased significantly after the therapy (1.3 +/- 1.1 vs. 2.7 +/- 1.2, P = 0.004). CONCLUSIONS: Serum levels of sIL-2R and sCD154 could be useful as indicators of inflammation and sRANKL/OPG ratios as markers of osteolytic activity in LCH patients. PMID- 16358319 TI - New phenotypes of functional expression of the mKir2.1 channel in potassium efflux-deficient Saccharomyces cerevisiae strains. AB - The functional expression of the mouse Kir2.1 potassium channel in yeast cells lacking transport systems for potassium and sodium efflux (ena1-4delta nha1delta) resulted in increased cell sensitivity to high external concentrations of potassium. The phenotype depended on the level of Kir2.1 expression and on the external pH. The activity of Kir2.1p in the yeast cells was almost negligible at pH 3.0 and the highest at pH 7.0. Kir2.1p was permeable for both potassium and rubidium cations, but neither sodium nor lithium were transported via the channel. Measurements of the cation contents in cells confirmed the higher concentration of potassium in cells with Kir2.1p. Specific inhibition of the mKir2.1 channel activity by Ba2+ cations was observed. The use of a mutant strain lacking both potassium efflux and uptake transporters (ena1-4delta nha1delta trk1delta trk2delta) enabled the monitoring of channel activity on two levels- the provision of the necessary amount of intracellular K+ in media with low potassium concentrations, and simultaneously, the channel's contribution to cell potassium sensitivity in the presence of high external K+. This combination of mutations proved to be a new, sensitive and practical tool for characterizing the properties of heterologously expressed transporters mediating both the efflux and influx of alkali-metal-cations. PMID- 16358322 TI - The YIG1 (YPL201c) encoded protein is involved in regulating anaerobic glycerol metabolism in Saccharomyces cerevisiae. AB - Under anaerobic conditions S. cerevisiae produces glycerol to regenerate NAD(+) from the excess NADH produced in cell metabolism. We here report on the role of an uncharacterized protein, Yig1p (Ypl201cp), in anaerobic glycerol production. Yig1p was previously shown to interact in two-hybrid tests with the GPP1 and GPP2 encoded glycerol 3-phosphatase (Gpp), and we here demonstrate that strains overexpressing YIG1 show strongly decreased Gpp activity and content of the major phosphatase, Gpp1p. However, cells overexpressing YIG1 exhibited only slightly decreased GPP1 transcript levels, suggesting that Yig1p modulates expression on both transcriptional and post-transcriptional levels. In agreement with such a role, a GFP-tagged derivate of Yig1p was localized to both the cytosol and the nucleus. Deletion or overexpression of YIG1 did not, however, significantly affect growth yield or glycerol yield in anaerobic batch cultures, which is consistent with the previously proposed low flux control exerted at the Gpp level. PMID- 16358323 TI - Ambient air concentrations of pesticides used in potato cultivation in Prince Edward Island, Canada. AB - Ambient air concentrations of nine selected pesticides used in potato cultivation were investigated on Prince Edward Island by collecting samples during the summer of 1998 at three potato farm sites and one non-agricultural site. In 1999, air samples were collected at a single potato farm site during local application of pesticides. The fungicide chlorothalonil was the only pesticide detected in every sample in both years, even in samples with a duration as little as 5 h. In 1998, maximum (45-458 ng m(-3)) and mean (22-193 ng m(-3)) concentrations of chlorothalonil from composite 42 h samples were one to two orders of magnitude greater at agricultural sites than at the non-agricultural site (3.9 and 2.5 ng m(-3), respectively). Maximum and mean concentrations of chlorothalonil from combined 24 h samples at the farm site in 1999 were higher than those measured in 1998 (636 and 284 ng m(-3), respectively). The ubiquitous presence in air of relatively high concentrations of chlorothalonil in agricultural areas on Prince Edward Island is likely related to its repeated use on potato farms where fungicides account for 80-90% of pesticides applied. Eight of nine pesticides were detected at farm sites in 1998 and they are ranked by mean concentration from highest to lowest as follows: chlorothalonil, methamidophos, azinphos methyl, alpha-endosulfan, beta-endosulfan, pirimicarb, metobromuron, metribuzin, metalaxyl and fluazifop-P-butyl. Concentrations of metalaxyl, pirimicarb, metobromuron and fluazifop-P-butyl in air are among the first reported values for these pesticides. Five pesticides were detected at the Summerside farm in 1999, but only two fungicides (chlorothalonil and metalaxyl) were used locally, while the presence of metribuzin, alpha-endosulfan and methamidophos in air was not associated with local application. Evidence of pesticide drift was observed for chlorothalonil, alpha-endosulfan and methamidophos, and these pesticides were identified as being of high concern in terms of potential wildlife exposure on the Island. PMID- 16358324 TI - The effect of box shape on the dynamic properties of proteins simulated under periodic boundary conditions. AB - The effect of the box shape on the dynamic behavior of proteins simulated under periodic boundary conditions is evaluated. In particular, the influence of simulation boxes defined by the near-densest lattice packing (NDLP) in conjunction with rotational constraints is compared to that of standard box types without these constraints. Three different proteins of varying size, shape, and secondary structure content were examined in the study. The statistical significance of differences in RMSD, radius of gyration, solvent-accessible surface, number of hydrogen bonds, and secondary structure content between proteins, box types, and the application or not of rotational constraints has been assessed. Furthermore, the differences in the collective modes for each protein between different boxes and the application or not of rotational constraints have been examined. In total 105 simulations were performed, and the results compared using a three-way multivariate analysis of variance (MANOVA) for properties derived from the trajectories and a three-way univariate analysis of variance (ANOVA) for collective modes. It is shown that application of roto translational constraints does not have a statistically significant effect on the results obtained from the different simulations. However, the choice of simulation box was found to have a small (5-10%), but statistically significant effect on the behavior of two of the three proteins included in the study. PMID- 16358325 TI - Binding site-based classification of coronaviral papain-like proteases. AB - The coronavirus replicase gene encodes one or two papain-like proteases (termed PL1pro and PL2pro) implicated in the N-terminal processing of the replicase polyprotein and thus contributing to the formation of the viral replicase complex that mediates genome replication. Using consensus fold recognition with the 3D JURY meta-predictor followed by model building and refinement, we developed a structural model for the single PLpro present in the severe acute respiratory syndrome coronavirus (SCoV) genome, based on significant structural relationships to the catalytic core domain of HAUSP, a ubiquitin-specific protease (USP). By combining the SCoV PLpro model with comparative sequence analyses we show that all currently known coronaviral PLpros can be classified into two groups according to their binding site architectures. One group includes all PL2pros and some of the PL1pros, which are characterized by a restricted USP-like binding site. This group is designated the R-group. The remaining PL1pros from some of the coronaviruses form the other group, featuring a more open papain-like binding site, and is referred to as the O-group. This two-group, binding site-based classification is consistent with experimental data accumulated to date for the specificity of PLpro-mediated polyprotein processing and PLpro inhibition. It also provides an independent evaluation of the similarity-based annotation of PLpro-mediated cleavage sites, as well as a basis for comparison with previous groupings based on phylogenetic analyses. PMID- 16358326 TI - Plasma membrane calcium ATPase is concentrated in the head of sea urchin spermatozoa. AB - Plasma membrane Ca2+ATPases (PMCAs) export Ca2+ from cells in a highly regulated manner, providing fine-tuning to the maintenance of intracellular Ca2+ concentrations. There are few studies of PMCAs in spermatozoa, which is surprising considering the importance of this enzyme in all cell types. Here we describe the primary structure and localization of the PMCA of sea urchin spermatozoa (suPMCA). The suPMCA is 1,154 amino acids and has 56% identity and 76% similarity to all 4 human PMCA isoforms. The suPMCA shares the features of a typical PMCA, including domains for calmodulin binding, ATP binding, ATPase phosphorylation, and 10 putative transmembrane segments with two large cytoplasmic loops. Southern blots show that suPMCA is a single copy gene. Treatment of live sea urchin sperm with the PMCA inhibitor, 5-(-6)-carboxyeosin, results in elevations of intracellular Ca2+ and loss of flagellar motility. Immunoblotting and immunoflorescence show that suPMCA is concentrated in the sperm head plasma membrane. In previous work, we showed that a plasma membrane K+ dependent Na+/Ca2+ exchanger (suNCKX), which also keeps Ca2+ low in these cells, is concentrated in the sperm flagellum. Thus, the sperm head and flagellum localize different gene products, both functioning to keep intracellular Ca2+ low, while the sperm swims in seawater containing 10 mM Ca2+. PMID- 16358327 TI - Impact of cis-proline analogs on peptide conformation. AB - The beta-turn is a common motif in both proteins and peptides and often a recognition site in protein interactions. A beta-turn of four sequential residues reverses the direction of the peptide chain and is classified by the phi and psi backbone torsional angles of residues i + 1 and i + 2. The type VI turn usually contains a proline with a cis-amide bond at residue i + 2. Cis-proline analogs that constrain the peptide to adopt a type VI turn led to peptidomimetics with enhanced activity or metabolic stability. To compare the impact of different analogs on amide cis-trans isomerism and peptide conformation, the conformational preference for the cis-amide bond and the type VI turn was investigated at the MP2/6-31+G** level of theory in water (polarizable continuum water model). Analogs stabilize the cis-amide conformations through different mechanisms: (1) 5 alkylproline, with bulky hydrocarbon substituent on the C(delta) of proline, increases the cis-amide population through steric hindrance between the alkyl substituent and the N-terminal residues; (2) oxaproline or thioproline, the oxazolidine- or thiazolidine-derived proline analog, favors interactions between the dipole of the heterocyclic ring and the preceding carbonyl oxygen; and (3) azaproline, containing a nitrogen atom in place of the C(alpha) of proline, prefers the cis-amide bond by lone-pair repulsion between the alpha-nitrogen and the preceding carbonyl oxygen. Preference for the cis conformation was augmented by combining different modifications within a single proline. Azaproline and its derivatives are most effective in stabilizing cis-amide bonds without introducing additional steric bulk to compromise receptor interactions. PMID- 16358328 TI - A self-stabilized model of the chymotrypsin catalytic pocket. The energy profile of the overall catalytic cycle. AB - A model of the catalytic triad of chymotrypsin is built assuring the arrangement and properties as they are within the complete enzyme. The model contains 18 amino acid residues of chymotrypsin and its substrate. A total of 135 atoms (including 70 heavy atoms) were subjected to full ab initio geometry optimizations through 127 individual steps along the reaction coordinate of the complete catalytic mechanism. It was shown that the described model of the catalytic apparatus forms a self-stabilized molecule ensemble without the rest of the enzyme and substrate. According to the calculations, the formations of the first and second tetrahedral intermediates in the model have 20.3 and 15.7 kcal/mol activation energy barriers, respectively. Removing elements of the catalytic apparatus such as the (1) catalytic aspartate or (2) the anion hole, as well as (3) inserting a water molecule "early" in the catalytic process, or (4) introducing conformational rigidity of the substrate, results in an increase of the above energy barrier of the first catalytic step in the model by 6.4, 13.7, 3.7, and 4.1 kcal/mol, respectively. Based on the calculated process one can conclude that the catalytic reaction in this model is much more similar to the reaction in the enzyme than to the reference reaction. To our knowledge, this is the first model system that mimics the complete catalytic mechanism. PMID- 16358329 TI - Contralateral hand anesthesia transiently improves poststroke sensory deficits. AB - OBJECTIVE: To test a possible strategy to alleviate somatosensory deficits after stroke. METHODS: Here, we applied ischemic nerve block to the intact hand of patients with chronic stroke, which in healthy subjects elicits improvements in sensibility of the other hand. RESULTS: We found that sensibility in the affected hand improved with intact hand anesthesia, but not with intact foot anesthesia or no anesthesia. INTERPRETATION: We conclude that reduction of sensory input from the intact hand leads to site-specific improvements in tactile discriminative skills in the affected hand after the period of anesthesia, a potentially relevant finding in designing neurorehabilitative interventions. PMID- 16358330 TI - Lower serotonin transporter binding in caudate in alcoholics. AB - Dorsal striatum is regulated by the serotonergic system, and it is a brain area with a role in the development of obsessive thought patterns, which may be related to addiction. In this study, possible alterations of [(3)H]citalopram binding to serotonin transporter (SERT) were evaluated in the dorsal striatum of Cloninger type 1 and 2 alcoholics, and nonalcoholic control subjects by postmortem whole-hemisphere autoradiography in humans. The SERT binding was significantly lower (-26%, effect size 1.74) in the caudate body of alcoholics. The SERT binding tended to be lower also in the other parts of the dorsal striatum in alcoholics, but the results did not reach significance. In addition, there was a significant positive correlation, especially in type 1 alcoholics, between the SERT binding in the body of the caudate and in the perigenual anterior cingulate cortex, an area in which the SERT binding has been shown to be lower among alcoholics. These results give preliminary evidence to suggest that the SERT binding in the dorsal striatum may be lower in alcoholics, and that the serotonergic system may be affected in cortical and striatal areas simultaneously. The cortico-striatal-thalamic axis may have an important role in fully developed addictions and the characterization of these correlations within the serotonergic system may lead to a better understanding of the anatomical dynamics underlying the neurochemistry of alcoholism. PMID- 16358331 TI - Effect of reduction in endogenous dopamine on extrastriatal binding of [11C]FLB 457 in rat brain--an ex vivo study. AB - Carbon-11 labeled FLB 457 has been used successfully as a selective, high affinity PET ligand for the quantification of extrastriatal D2-like receptors in man. This study was carried out in rats to investigate regional values for maximal binding and ED50 (a measure of apparent K(d)) for the radioligand in vivo in control animals and in a group pretreated with the neuronal impulse flow inhibitor, gamma-butyrolactone. The aims were to obtain further information regarding the specific activity needed to ensure tracer kinetics and to investigate baseline occupancy by dopamine (DA), each relevant to optimal clinical use of the radioligand. Regional B(max) values were consistent with the distribution of D2-like receptors in rat brain. Of interest, 60% of the binding in cerebellum, often used as a low-binding "reference region" for PET quantification, was saturable, with B(max) only 2- to 3-fold less than that in neocortex, hippocampus, and thalamus. ED50 values were in the range 2-3 nmol/kg, confirming minimal receptor occupancy by the tracer in human PET, using high but achievable specific activities. In the majority of extrastriatal tissues, reduction in synaptic DA did not significantly decrease the apparent K(d), except in cortical regions, where the extent of the effect suggested a low ( approximately 10%), but measurable baseline receptor occupancy by DA. PMID- 16358332 TI - Low mRNA levels of RGS4 splice variants in Alzheimer's disease: association between a rare haplotype and decreased mRNA expression. AB - Regulator of G-protein signaling 4 (RGS4) showed decreased mRNA levels in Alzheimer's disease in a large collection of human brain autopsies from prefrontal cortex. The expression levels of three RGS4 splice variants were examined in the same samples, and the association between RGS4 gene expression and/or the disease with single nucleotide polymorphisms located in this gene was explored. We show that all splice variants are down-regulated in patients. We also demonstrate that one rare haplotype (ATAG) is associated with decreased mRNA levels in both cases and controls. Our results suggest that an altered regulation in transcription initiation may be an important mechanism for low RGS4 protein levels in Alzeimer's disease. PMID- 16358333 TI - Mutant androgen receptor accumulation in spinal and bulbar muscular atrophy scrotal skin: a pathogenic marker. AB - OBJECTIVE: Spinal and bulbar muscular atrophy (SBMA) is a hereditary motor neuron disease caused by the expansion of a polyglutamine tract in the androgen receptor (AR). The nuclear accumulation of mutant AR is central to the pathogenesis of SBMA. Androgen deprivation with leuprorelin inhibits mutant AR accumulation, resulting in rescue of neuronal dysfunction in a mouse model of SBMA. This study aimed to investigate whether mutant AR accumulation in the scrotal skin is an appropriate biomarker of SBMA. METHODS: Immunohistochemistry of both scrotal skin and the spinal cord was performed on five autopsied SBMA cases. Neurological severity and scrotal skin findings were studied in another 13 patients. Five other patients received subcutaneous injections of leuprorelin and underwent scrotal skin biopsy. RESULTS: The degree of mutant AR accumulation in scrotal skin epithelial cells tended to be correlated with that in the spinal motor neurons in autopsy specimens, and it was well correlated with CAG repeat length and inversely correlated with the amyotrophic lateral sclerosis functional scale. Leuprorelin treatment inhibited mutant AR protein accumulation in the scrotal skin of SBMA patients. INTERPRETATION: These observations suggest that scrotal skin biopsy findings are a potent pathogenic marker of SBMA and can be a surrogate end point in therapeutic trials. PMID- 16358334 TI - Apolipoprotein E and alpha brain rhythms in mild cognitive impairment: a multicentric electroencephalogram study. AB - OBJECTIVE: Relationships between the apolipoprotein E epsilon4 allele and electroencephalographic (EEG) rhythmicity have been demonstrated in Alzheimer's disease (AD) patients but not in the preclinical stage prodromic to it, namely, mild cognitive impairment (MCI). The present multicentric EEG study tested the hypothesis that presence of epsilon4 affects sources of resting EEG rhythms in both MCI and AD subjects. METHODS: We enrolled 89 MCI subjects (34.8% with epsilon4) and 103 AD patients (50.4% with epsilon4). Resting eyes-closed EEG data were recorded for all subjects. EEG rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5-13Hz), beta 1 (13-20 Hz), and beta 2 (20-30 Hz). EEG cortical sources were estimated by low-resolution brain electromagnetic tomography. RESULTS: Results showed that amplitude of alpha 1 and 2 sources in occipital, temporal, and limbic areas was lower in subjects carrying the epsilon4 allele than in those not carrying the epsilon4 allele (p < 0.01). This was true for both MCI and AD. For the first time to our knowledge, a relationship was shown between ApoE genotype and global neurophysiological phenotype (ie, cortical alpha rhythmicity) in a preclinical AD condition, MCI, in addition to clinically manifest AD. INTERPRETATION: Such a demonstration motivates future genotype-EEG phenotype studies for the early prediction of AD conversion in individual MCI subjects. PMID- 16358335 TI - Clinical heterogeneity of alpha-synuclein gene duplication in Parkinson's disease. AB - OBJECTIVE: Recently, genomic multiplications of alpha-synuclein gene (SNCA) have been reported to cause hereditary early-onset parkinsonism. The objective of this study was to assess the frequency of SNCA multiplications among autosomal dominant hereditary Parkinson's disease (ADPD). METHODS: We screened 113 ADPD probands and 200 sporadic PD cases by quantitative polymerase chain reaction and confirmed SNCA multiplications by fluorescence in situ hybridization (FISH) and comparative genomic hybridization array. RESULTS: Two families (two patients from Family A and one from Family B) with SNCA duplication were identified among ADPD patients. Even though they had the same SNCA duplication, one patient had dementia. Because there was exactly the same difference between the regions originated from each patient, the finding suggests that the phenotype of SNCA multiplication may be also influenced by the range of duplication region. We also detected asymptomatic carriers in the families of both patients. Interestingly, the penetrance ratio was 33.3% (2/6) in one kindred, indicating that the ratio was very much lower than expected. INTERPRETATION: These two newly identified Japanese patients with SNCA duplication and the five previously identified American and European families with SNCA triplication or duplication mutations indicate that the incidence of SNCA multiplication may be more frequent than previously estimated. PMID- 16358336 TI - Motor neuron disease in a patient with a mitochondrial tRNAIle mutation. AB - OBJECTIVE: Motor neuron disease (MND) is a common neurodegenerative condition for which the underlying cause is uncertain in many patients. We identified a patient with clinical features suggestive of MND but additional cardiac and metabolic symptoms. We wished to determine if the clinical features were due to a mitochondrial DNA mutation. METHODS: The brain and spinal cord were studied using neuropathological techniques and agenetic defect investigated in individual neurons. RESULTS: There were atypical neuropathological features and genetic studies identified a pathogenic, heteroplasmic mitochondria tRNA(Ile) (4274T>C) mutation. INTERPRETATION: This case adds to the phenotypic variation seen in mitochondrial DNA disease but also highlights the potential role of mitochondrial dysfunction in the cause of MND. PMID- 16358337 TI - Gene regulation by IL-1beta independent of IL-1R1 in the mouse brain. AB - Interleukin-1 (IL-1) is a key pro-inflammatory cytokine that has diverse actions in the brain as a regulator of host defense responses and a mediator of inflammation. Two major agonists, IL-1alpha and IL-1beta, bind to a single known functional type-1 IL-1 receptor (IL-1RI) that associates with the accessory protein (IL-1RAcP), resulting in signal transduction. However, recent evidence suggests that some actions of IL-1 in the brain may be independent of IL-1R1 and the classical IL-1 signaling pathways, pointing to an as-yet unidentified functional receptor for IL-1. In this study, we have used cDNA microarray-based gene expression profiling to identify the possible genes induced by IL-1beta independently of IL-1R1. IL-1beta induced potential changes (greater than 2-fold vs. vehicle-treated) in the expression of up to 1285 candidate genes in wild-type primary mixed glia, and 404 candidate genes in IL-1R1-/- cells of the same type. Real-time quantitative polymerase chain reaction (PCR) on selected genes revealed that pentraxin-3, was upregulated by IL-1beta in wild-type, but not in IL-1R1-/- mixed glia. Amongst the other genes for which expression was modified by IL-1beta in IL-1R1-/- cells, we selected alpha-syntrophin and demonstrated by real-time quantitative PCR that expression of this gene is significantly downregulated by IL-1beta in primary mixed glia prepared from wild-type, IL-1R1-/-, IL-1RAcP-/- or MyD88-/- mice. In contrast, IL-1alpha fails to downregulate alpha-syntrophin expression in wild-type or IL-1R1-/- mixed glia. These results show that IL-1beta exclusively downregulates alpha-syntrophin expression independently of IL-1R1, and suggest the expression of additional functional IL-1 receptors in the CNS. PMID- 16358338 TI - Allele C-specific methylation of the 5-HT2A receptor gene: evidence for correlation with its expression and expression of DNA methylase DNMT1. AB - Differential DNA methylation has been suggested to contribute to differential activity of alleles C and T and thereby to genetic associations between the C/T(102) polymorphism in the 5-HT2A receptor gene (5HT2AR) and psychiatric disorders. We surveyed methylation in two CpG sites, which are specific to allele C. The majority of allele C-specific CpG sites were methylated in human temporal cortex and peripheral leukocytes and levels of methylation varied between individuals. Levels of methylation in the promoter correlated significantly with the expression of 5HT2AR. Methylation of allele C-specific CpG sites in the first exon correlated significantly with the expression of DNA methylase 1 (DNMT1) but not S-adenosylhomocysteine hydrolase (AHCY). These findings support the hypothesis that allele-specific DNA methylation is involved in regulation of 5HT2AR expression, influencing expression differences between alleles C and T. PMID- 16358339 TI - Neurodegenerative actions of interleukin-1 in the rat brain are mediated through increases in seizure activity. AB - The cytokine interleukin-1 (IL-1) is an established and important mediator of diverse forms of neuronal injury in experimental animals. However, its mechanisms of action remain largely unknown. We have reported previously that IL-1 markedly enhances excitotoxic injury induced in the rat by striatal administration of the excitotoxin alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), leading to widespread neuronal loss throughout the ipsilateral cortex. Here we tested the hypothesis that IL-1 causes this injury through induction and/or enhancement of seizure activity in the rat. Consistently with this hypothesis, intrastriatal injection of AMPA or AMPA with IL-1 in the rat brain increased c-Fos expression in regions similar to those in which c-Fos has been reported previously in response to seizures. A significant increase in cortical neuronal activity (number of c-Fos positive cells) was observed in response to AMPA with IL-1 compared with AMPA (8 hr after injection). Increased seizure duration [3,522 +/- 660 sec (SEM) vs. 1,415 +/- 301 sec; P < 0.001] and cell death volume (140 +/- 20 mm3 vs. 52 +/- 6 mm3; P < 0.001) were seen in response to coinfusion of AMPA with IL-1 vs. AMPA alone. In addition, the anticonvulsant diazepam (intraperitoneal) significantly reduced cell death (P < 0.001) and seizure duration (P < 0.001) induced by AMPA with IL-1, and a significant correlation was found between seizure duration and cell death volume. These findings support our hypothesis that IL-1 enhances excitotoxic injury by enhancement of seizures, which may be of relevance to IL-1 actions in other forms of neuronal injury, including cerebral ischemia. PMID- 16358340 TI - Membranous obliterative bronchitis: a proposed unifying model. AB - Three adults with cystic fibrosis (one after lung transplantation) presented with fever, chest pain, and acute radiographic changes. The changes included a cavitary lesion of the lung, acute dense infiltrates, and lobar collapse. After failing conventional antibiotic therapy, the patients underwent flexible bronchoscopy. All had bronchial obstruction by a membrane that had completely occluded the bronchial orifice at the bifurcation of the bronchi. Therapeutic interventions ranged from continuing intravenous antibiotics, bronchoscopy assisted perforation of the membrane by sharp instrumentation, and transthoracic needle-guided perforation of the membrane with subsequent stenting of the orifice. The patients recovered, but the posttransplant patient had recurrent membranous obstructions with multiple interventions. The cause and triggers of the process are unknown. Based on repeated observations of the evolution of the membranes, and histologic material from bronchoscopies, we propose a putative paradigm of the natural history of the process. We suggest that local stimuli generate a richly vascularized granulation polyp, which progresses in a "shutter like" motion to form partial or completely obstructive membranes. The subsequent course depends on the vascular supply to the membrane. We also propose that similar processes may be the underlying pathologic events in some cases of lung abscess and necrotizing pneumonia. PMID- 16358341 TI - Reduced exhaled nitric oxide in children after testing of maximal expiratory pressures. AB - Spirometry in adult subjects can induce a fall in concentration of exhaled nitric oxide (FE(NO)). Scarce information is available on the FE(NO) decrease after spirometry or after other forced lung-function maneuvers in children. We compared changes in FE(NO) induced by repeated spirometry and testing of maximal expiratory pressures (P(Emax)). Twenty-four sex- and age-matched children aged 9 18 years (mean age +/- SD, 13.3 +/- 2.8 years; 12 healthy, 12 asthmatic) were allocated to 1-week-apart sessions of repeated maneuvers of either forced vital capacity (FVC) or P(Emax). Baseline FE(NO) measurements were followed by FVC or P(Emax) maneuvers every 15 min for 45 min, whereas FE(NO) was measured at each step for 60 min. After repeated P(Emax) but not after FVC maneuvers, FE(NO) values decreased significantly from baseline in both groups. In healthy children, geometric mean FE(NO) (95% confidence intervals) decreased from 9.1 (7.0-11.8) ppb at baseline to 8.2 (6.3-10.6) ppb at 15 min and 7.7 (5.6-10.6) ppb at 30 min (P < 0.05 and P < 0.01, respectively), and remained unchanged at 45 and 60 min. In asthmatic children, FE(NO) levels fell from 21.6 (13.3-34.9) ppb at baseline to 15.1 (9.1-25.1) ppb at 15 min and remained low at 30, 45, and 60 min: 17.8 (10.7-29.5) ppb, 17.5 (10.2-30.1) ppb, and 17.6 (10.6-29.2) ppb, P < 0.01, for all differences from baseline. Repeated P(Emax) and FVC maneuvers increased FE(NO) variability, as compared with repeated FE(NO) measurements alone. Previous forced lung-function maneuvers may affect FE(NO) measurements in children. Although P(Emax) testing has a greater influence than spirometry on FE(NO) levels in children, both procedures should be avoided before measuring FE(NO). PMID- 16358342 TI - Evidence of ventilatory constraints in healthy exercising prepubescent children. AB - We assessed expiratory airflow limitation (exp FL) in 18 healthy prepubescent children (6 girls and 12 boys, 10.1 +/- 0.3 years old), and examined how it might modulate regulation of tidal volume (V(T)) during exercise. The children performed a maximal incremental exercise on a cycle ergometer, preceded and followed by pulmonary function tests. Throughout exercise, breathing flow-volume loops were plotted into the maximal flow-volume loop (MFVL) measured at rest. End expiratory and end-inspiratory lung volumes were estimated by measuring expiratory reserve volume relative to forced vital capacity (ERV/FVC), and inspiratory reserve volume relative to forced vital capacity (IRV/FVC), respectively. The exp FL, expressed as a percentage of V(T), was defined as the part of the tidal breath meeting the boundary of the MFVL. Ten children (FL) presented an exp FL at peak exercise (range, 16-78% of V(T)), and the remaining 8 constituted a non-flow-limited group (NFL). At peak exercise, FL presented a higher IRV/FVC and lower ERV/FVC (P < 0.01) than NFL children, demonstrating two different exercise breathing patterns. These results suggest that the NFL regulated V(T) at high lung volume, avoiding exp FL, while the FL breathed at low lung volume, leading to exp FL. At peak exercise, FL presented lower values of minute ventilation (P<0.05) and oxygen uptake (P<0.05) than NFL. Nevertheless, oxygen arterial saturation and dyspnea were similar in the two groups. In conclusion, ventilatory constraints may occur in healthy prepubescent children and result in relative dynamic hyperinflation or expiratory flow limitation. PMID- 16358344 TI - Interaction of bovine serum albumin with chrysotile: spectroscopic and morphological studies. AB - The biodurability of chrysotile fibers, which is related to their cytotoxicity and mutagenic responses, is strongly affected by the surface chemical adsorption of biological molecules. Natural chrysotile is a heterogeneous material in both structure and composition. The availability of synthetic stoichiometric chrysotile of constant structure and uniform morphology has allowed us to investigate its interaction with bovine serum albumin (BSA). By using transmission electron microscopy (TEM) and atomic force microscopy (AFM), we have obtained the first morphological evidence of albumin adsorption onto chrysotile nanocrystals. FTIR spectroscopy was used to quantify modifications of BSA secondary structure that were induced by the surface interaction. The protein transition to beta-turns allows a stronger interaction between the protein hydrophilic side-chains and the charged asbestos surface, which is consistent with hydrogen bonds involving the superficial OH groups. Synthetic stoichiometric chrysotile nanocrystals were shown to be an ideal reference standard with which to study the interaction of asbestos fibers with biological systems, in order to elucidate the chemical mechanisms of asbestos toxicity. PMID- 16358345 TI - Computational study of solvent effects and the vibrational spectra of Anderson polyoxometalates. AB - The structures and vibrational frequencies of the type II Anderson heteropolyanions [TeMo6O24]6- and [IMo6O24]5- have been calculated by using density functional theory using a number of common functionals and basis sets. For the first time, Raman intensities have been calculated and the effect of solvent on the modeling has been investigated. The calculated IR and Raman spectral traces are in good agreement with experiment allowing the characteristic group frequencies for this class of polyoxometalate to be identified. The stretching vibrations of the molybdenum-oxygen bonds are predicted to occur at somewhat lower frequencies than in the type I polyoxometalates. Stretching of the heteroatom-oxygen bonds occurs at significantly lower frequencies than in the Keggin anions as a simple consequence of the higher coordination number of the central heteroatom in the Anderson systems. For the [Mo2O7]2- and [Mo6O19]2- ions, the relatively low negative charge leads to small structural changes when solvent is included. In these systems, solvent leads to an increase in the bond polarity and a decrease in the covalent bond orders, resulting in decreases in the calculated frequencies. For the Anderson anions, the higher negative charges leads to greater solvent effects with contraction of the clusters and increases in the frequencies of bands due to stretching of the two, cis-related molybdenum oxygen bonds. PMID- 16358343 TI - Shrinking lung syndrome in a 14-year-old boy with systemic lupus erythematosus. AB - Pulmonary complications occur frequently in people with systemic lupus erythematosus. We report on an adolescent with an acute onset of dyspnea and pleuritic chest pain with severe restrictive lung physiology on pulmonary function testing (forced vital capacity, 20% of predicted) who had no evidence of parenchymal lung or pleural disease. He was found to have restricted diaphragmatic movement as assessed by fluoroscopy, without evidence of generalized respiratory muscle weakness. His clinical presentation and results of diagnostic tests were typical for shrinking lung syndrome. Given the rarity of shrinking lung syndrome in the pediatric age range, many clinicians are not aware of it as a clinical entity. Shrinking lung syndrome should be included in the differential diagnosis of dyspnea in both children and adults with systemic lupus erythematosus. PMID- 16358346 TI - Enhanced solubilization of bovine serum albumin in reverse micelles by compressed CO2. AB - The effect of compressed CO2 on the solubilization of bovine serum albumin (BSA) in water/sodium bis-(2-ethylhexyl) sulfosuccinate (AOT)/isooctane reverse micelles was studied by observing phase behavior and recording UV-visible spectra under different conditions. The pH values within the water cores of reverse micelles at different CO2 pressures were also determined. The solubilization capacity of the reverse micelles for the protein increased considerably as CO2 pressure increased within the low-pressure range, but decreased at higher CO2 pressures, so that the micelles eventually lost their ability to solubilize the protein. The effect of CO2 on the stability of the reverse micelles played an important role in the relationship between pressure and protein solubility. A "multicomplex" model was proposed to explain these effects. The different solublization capacities within different pressure ranges demonstrates the unique advantage of using compressed CO2 in the extraction of proteins with reverse micelles. PMID- 16358347 TI - Facile "green" synthesis, characterization, and catalytic function of beta-D glucose-stabilized Au nanocrystals. AB - We present a straightforward, economically viable, and "green" approach for the synthesis and stabilization of relatively monodisperse Au nanocrystals with an average diameter of 8.2 nm (standard deviation, SD=2.3 nm) by using nontoxic and renewable biochemical of beta-D-glucose and by simply adjusting the pH environment in aqueous medium. The beta-D-glucose acts both as reducing agent and capping agent for the synthesis and stabilization of Au nanocrystals in the system. The UV/Vis spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, transmission electron microscopy (TEM), electron diffraction (ED), and X-ray diffraction (XRD) techniques were employed to systematically characterize Au nanocrystals synthesized. Additionally, it is shown that these beta-D-glucose stabilized Au nanocrystals function as effective catalyst for the reduction of 4 nitrophenol in the presence of NaBH4 (otherwise unfeasible if only the strong reducing agent NaBH4 is employed), which was reflected by the UV/Vis spectra of the catalytic reaction kinetics. PMID- 16358348 TI - A synthetic camel anti-lysozyme peptide antibody (peptibody) with flexible loop structure identified by high-resolution affinity mass spectrometry. AB - We describe the synthesis and characterisation of the fully functional molecular recognition structure of a 26-amino acid residue peptide antibody, referred to as peptibody, designed from a monoclonal single-domain antibody fragment derived from a camel heavy-chain antibody. The CDR3 region (CDR = complementarity determining region) of the cAbLys3 camel antibody fragment, which binds to the active site of hen eggwhite lysozyme (HEL) and acts as a potent enzyme inhibitor by mimicking an oligosaccharide substrate, was prepared by solid-phase peptide synthesis. To obtain a closed loop-like structure resembling that in the crystal structure, N- and C-terminal cysteine residues were added to the linear peptide and oxidised to a cyclic disulfide-bridged peptide by using dimethylsulfoxide. A further, internal cysteine-12 residue was acetamidomethyl-protected to prevent possible oxidative byproducts. Affinity separation on a lysozyme microcolumn combined with MALDI-TOF mass spectrometry revealed that the peptide resumed high affinity to lysozyme only after deprotection of Cys-12, suggesting the importance of this paratope sequence for epitope recognition. The complex of lysozyme and active peptibody was characterised directly by conducting high-resolution ESI FTICR mass spectrometry, which provided a molecular comparison of affinities for linear and cyclic peptibodies. PMID- 16358349 TI - Electrochemistry and photophysics of donor-substituted triarylboranes: symmetry breaking in ground and excited state. AB - We synthesized a series of amino substituted triarylboranes (TABs) 1-3 by copper(I)-catalyzed cross-coupling reactions. The title compounds were investigated by means of cyclic voltammetry (CV) and UV-visible absorption and fluorescence spectroscopy. Electrochemical oxidation of tris(4-carbazolyl-2,6 dimethylphenyl)borane (3) leads to the formation of an electroactive polymer film on the electrode surface. The charge-transfer (CT) absorption band of all three TABs shows a pronounced negative solvatochromism, while the emission is positively solvatochromic. By combining Jortner's theory, AM1 computations, and electrooptical absorption measurements (EOAM), this unexpected behavior was shown to be due to a dipole inversion upon S0-->S1 excitation. Furthermore, polarized steady-state fluorescence spectroscopy and EOAM prove that the ground-state geometry of 3 is of lower symmetry than D3 and that the excitation energy can be transferred from one subchromophore to another within the lifetime of the excited state. Exciton-coupling theory was used to quantitatively analyze this excitation transfer. PMID- 16358350 TI - One-electron redox processes during polyoxometalate-mediated photocatalytic reactions of TiO(2) studied by two-color two-laser flash photolysis. AB - The one-electron redox processes of several compounds during polyoxometalate (POM)-mediated photocatalytic reactions of TiO(2) were investigated using the two color two-laser flash-photolysis technique. The efficiency of the one-electron oxidation of aromatic sulfides by the trapped hole (h(tr) (+)) or the surface bound OH radical (OH(s) (.)) is found to be significantly enhanced due to electron transfer from the conduction band (CB) of TiO(2) to the POM. The efficiency of the electron transfer from the CB of TiO(2) to the POM decreases in the order H(2)W(12)O(40) (6-) < SiW(12)O(40) (4-) < PW(12)O(40) (3-), that is, it depends on the reduction potential (E(red)) of the POMs. Electron injection from PW(12)O(40) (4-) in the excited state (PW(12)O(40) (4-*)) to the CB of TiO(2) was clearly observed using the two-color two-laser flash-photolysis technique. Storage of electrons in the TiO(2)/PW(12)O(40) (3-)/methyl viologen (MV(2+)) ternary system was also achieved upon two-color two-laser irradiation. PMID- 16358351 TI - Gold(I)-catalyzed cyclizations of 1,6-enynes: alkoxycyclizations and exo/endo skeletal rearrangements. AB - Gold(I) complexes are the most active catalysts for alkoxy- or hydroxycyclization and for skeletal rearrangement reactions of 1,6-enynes. Intramolecular alkoxycyclizations also proceed efficiently in the presence of gold(I) catalysts. The first examples of the skeletal rearrangement of enynes by the endocyclic cyclization pathway are also documented. Iron(III) is also able to catalyze exo and endo skeletal rearrangements of 1,6-enynes, although the scope of this transformation is more limited. The gold(I)-catalyzed endocyclic cyclization proceeds by a mechanism different from those followed in the presence of PdII, HgII, or RhI catalysts. PMID- 16358352 TI - Gold(I)-catalyzed intramolecular cyclopropanation of dienynes. AB - Gold(I) complexes are the most active catalysts for the biscyclopropanation of dienynes to form tetracyclic compounds. PtII and ZnII are also able to promote the biscyclopropanation, although less efficiently. The configurations obtained in all cases with the use of gold(I) catalysts can be explained by the pathway proceeding through anti cyclopropyl gold carbenes. Similar intermediates are most probably involved in reactions catalyzed by RuII and PtII. Two different cyclopropanation pathways have been found; they depend on the structures of the cyclopropyl gold carbenes (anti or syn) and the relative arrangements of the metal carbenes and the alkenes. PMID- 16358353 TI - The conformers of phenylglycine. AB - The neutral form of the unnatural amino acid phenylglycine was vaporized by laser ablation, and the presence of two conformers was detected in a supersonic expansion by Fourier transform microwave spectroscopy. Both conformers were unequivocally identified by comparison of their experimental rotational and quadrupole coupling constants with those calculated ab initio. The most stable conformer is stabilized by intramolecular hydrogen bonds N-H...O=C, N-H...pi (with the closest C-C bond in the aromatic ring), and a cis-COOH interaction. The other conformer exhibits a O-H...N hydrogen bond between the hydrogen atom of the hydroxyl group and the lone pair at the nitrogen atom. PMID- 16358354 TI - Progress towards interrupting intrauterine transmission of cytomegalovirus? PMID- 16358356 TI - Methylation analysis of hMLH1 gene promoter by a bisulfite-sensitive single strand conformation polymorphism-capillary electrophoresis method. AB - DNA methylation is an important epigenetic modification that alters transcription in those genes containing CpG islands. In this report, a novel DNA methylation analysis method was developed employing bisulfite-single strand conformation polymorphism combined with capillary electrophoresis (bisulfite-SSCP-CE). During the bisulfite treatment of genomic DNA, a high concentration of sodium bisulfite (4.8 mol/L) was preferred in order to shorten reaction time and minimize template degradation. The methylated and unmethylated ssDNA of hMLH1 promoter were simultaneously separated under the optimized CE conditions, including 6% SLPA with 10% glycerol as sieving medium, 25 degrees C as separation temperature and 12 kV as running voltage. The heterogeneous methylation of hMLH1 promoter was identified in 13 of 64 colorectal cancer patients. Moreover, hMLH1 promoter methylation had a significant relationship with protein expression loss and increased with the age of patients. Our results indicated that DNA methylation analysis for a large number of clinical samples would be facilitated by use of the bisulfite-SSCP-CE method. PMID- 16358357 TI - Optical fiber light-emitting diode-induced fluorescence detection for capillary electrophoresis. AB - A highly sensitive optical fiber light-emitting diode (LED)-induced fluorescence detector for CE has been constructed and evaluated. In this detector, a violet or blue LED was used as the excitation source and an optical fiber with 40 microm OD was used to transmit the excitation light. The upper end of the fiber was inserted into the separation capillary and was situated right at the detection window. Fluorescence emission was collected by a 40 x microscope objective, focused on a spatial filter, and passed through a cutoff filter before reaching the photomultiplier tube. Output signals were recorded and processed with a computer using in-house written software. The present CE/fluorescence detector deploys a simple and inexpensive optical system that requires only an LED as the light source. Its utility was successfully demonstrated by the separation and determination of amino acids (AAs) labeled with naphthalene-2,3-dicarboxaldehyde (NDA) and FITC. Low detection limits were obtained ranging from 17 to 23 nM for NDA-tagged AAs and 8 to 12 nM for FITC-labeled AAs (S/N=3). By virtue of such valuable features as low cost, convenience, and miniaturization, the presented detection scheme was proven to be attractive for sensitive fluorescence detection in CE. PMID- 16358358 TI - Mutations in mitochondrial-encoded cytochrome c oxidase subunits I, II, and III genes detected in Alzheimer's disease using single-strand conformation polymorphism. AB - A "mitochondrial hypothesis" of late onset Alzheimer's disease (AD) has been proposed. Biochemical studies indicate that there is a significant decrease in cytochrome oxidase (CO) activity as well as perturbed CO I and CO III mRNA levels in platelets and brain tissue from Alzheimer's patients. Using the electrophoretic mutation detection technique SSCP and DNA sequencing, we have identified 20 point mutations in the mitochondrial-encoded CO subunits (CO I, II, and III) in AD and age-matched control brain samples. Eight of the mutations are new variants of the mitochondrial genome. The efficiency of SSCP in detecting mutations in the CO subunits was estimated to be 80% when compared to dideoxy sequencing. One of the mutations (at position 9,861) results in a phenylalanine- >leucine substitution at a highly conserved residue in CO III. CO activity was reduced by an average of 35% in all AD brains compared to age-matched control samples, which agrees with previous reports. CO activity in one of the AD brain samples carrying the 9,861 mutation decreased by 80% relative to control brain samples, suggesting that the phenotypic expression of this mutation may result in reduced CO activity and compromised mitochondrial function. PMID- 16358359 TI - Dynamics of Arabidopsis thaliana soluble proteome in response to different nutrient culture conditions. AB - In an effort to determine the best extraction procedure compatible with the high reproducible 2-DE, different methods of soluble protein extraction from Arabidopsis cell culture suspensions grown in Gamborg B5 medium were tested. A reference 2-DE map was established for this soluble extract revealing 1184 spots. The most abundant protein spots were excised, trypsin-digested, and mass spectra obtained via MALDI-TOF and/or LC coupled to ESI-MS. Three hundred and thirty one proteins were identified and their functions were defined based on sequence comparisons and classified in different protein families. In order to analyze the impact of culture medium on the Arabidopsis proteome, we performed the 2-DE map from Arabidopsis cell suspensions cultured in another growth medium Murashige and Skoog (M-S) and 327 major spots were identified. Using PDQuest imaging analysis, significant increases in the amount of several housekeeping enzymes, stress/defense proteins, and heat shock proteins were found in M-S medium. Modified expression of certain proteins and detection of new isoforms involved in nitrate assimilation, nitrogen, and sulfur metabolism were also observed in the M S medium. This study provides the first 2-DE maps of the soluble proteome of Arabidopsis cell suspensions. The comparative analysis of the Arabidopsis proteome in respect to different nutrient supplies shows that the culture medium may significantly influence the expression pattern of major soluble proteins in Arabidopsis cells. This work also constitutes an important step for further proteomic analysis concerning cell responses to abiotic or biotic stresses. PMID- 16358360 TI - IL-23 leads to diabetes induction after subdiabetogenic treatment with multiple low doses of streptozotocin. AB - IL-23, a proximal regulator of IL-17, may be a major driving force in the induction of autoimmune inflammation. We have used a model of subdiabetogenic treatment with multiple low doses of streptozotocin (MLD-STZ; 4 x 40 mg/kg body weight) in male C57BL/6 mice to study the effect of IL-23 on immune-mediated beta cell damage and the development of diabetes, as evaluated by blood glucose, quantitative histology, immunohistochemistry and expression of relevant cytokines in the islets. Ten daily injections of 400 ng IL-23, starting on the first day of MLD-STZ administration led to significant and sustained hyperglycemia along with weight loss compared with controls (no IL-23), and a significant increase in the number of infiltrating cells, a lower insulin content, enhanced apoptosis, expression of IFN-gamma and IL-17 (not seen in the controls) and a significant increase in the expression of TNF-alpha and IL-18 in the pancreatic islets. IL-23 treatment started 5 days prior to MLD-STZ administration had no effect on diabetogenesis or cytokines expression in the pancreatic islets. We provide the first evidence in an animal model that IL-23 is involved in the development of type-1 diabetes, by inducing IL-17 and possibly IFN-gamma production in the target tissue. PMID- 16358361 TI - Severe combined immunodeficiency and microcephaly in siblings with hypomorphic mutations in DNA ligase IV. AB - DNA double-strand breaks (dsb) during V(D)J recombination of T and B lymphocyte receptor genes are resolved by the non-homologous DNA end joining pathway (NHEJ) including at least six factors: Ku70, Ku80, DNA-PK(cs), Artemis, Xrcc4, and DNA ligase IV (Lig4). Artemis and Lig4 are the only known V(D)J/NHEJ factors found deficient in human genetic disorders. Null mutations of the Artemis gene result in a complete absence of T and B lymphocytes and increased cellular sensitivity to ionizing radiations, causing radiosensitive-SCID. Mutations of Lig4 are exclusively hypomorphic and have only been described in six patients, four exhibiting mild immunodeficiency associated with microcephaly and developmental delay, while two patient had leukemia. Here we report a SCID associated with microcephaly caused by compound heterozygous hypomorphic mutations in Lig4. Residual activity of Lig4 in these patients is underscored by a normal pattern of TCR-alpha and -beta junctions in the T cells of the patients and a moderate impairment of V(D)J recombination as tested in vitro. These observations contrast with the severity of the clinical immunodeficiency, suggesting that Lig4 may have additional critical roles in lymphocyte survival beyond V(D)J recombination. PMID- 16358362 TI - Regulatory T cells induce a privileged tolerant microenvironment at the fetal maternal interface. AB - The mechanisms underlying immune tolerance during pregnancy are poorly understood. In this regard, Treg seem to play an important role in mediating maternal tolerance to the fetus. We proposed a crucial role of T regulatory cells (Treg) in avoiding immunological rejection of the fetus after observing diminished number and function of Treg in abortion-prone mice. We further confirmed the protective role of Treg during pregnancy by transferring pregnancy induced Treg into abortion-prone mice, which prevented rejection. Here, we analyzed the mechanisms involved in Treg-mediated protection. As expected, Treg therapy prevented abortion, while expanding the peripheral and thymic Treg population. Surprisingly, the decidual levels of the Th1 cytokines IFN-gamma and TNF-alpha were not diminished after therapy. Interestingly, the mRNA levels of leukemia inhibitory factor, TGF-beta and heme oxygenase-1 at the fetal-maternal interface were dramatically up-regulated after Treg transfer, while the levels of indolamine 2,3-dioxygenase remained unchanged. Our data suggest that Treg treatment can not prevent T cell infiltration or high Th1 levels but is able to create a privileged tolerant microenvironment at the fetal-maternal interface, further shedding light onto the molecular mechanisms involved in pregnancy tolerance. PMID- 16358363 TI - PD-L1 and PD-L2 have distinct roles in regulating host immunity to cutaneous leishmaniasis. AB - To compare the roles of programmed death 1 ligand 1 (PD-L1) and PD-L2 in regulating immunity to infection, we investigated responses of mice lacking PD-L1 or PD-L2 to infection with Leishmania mexicana. PD-L1(-/-) and PD-L2(-/-) mice exhibited distinct disease outcomes following infection with L. mexicana. In comparison to susceptible WT mice, PD-L1(-/-) mice showed resistance to L. mexicana, as demonstrated by reduced growth of cutaneous lesions and parasite burden. In contrast, PD-L2(-/-) mice developed exacerbated disease with increased parasite burden. Host resistance to L. mexicana is partly associated with the development of a Th1 response and down-regulation of the Th2 response. Both PD L1(-/-) and PD-L2(-/-) mice produced levels of IFN-gamma similar to WT mice. However, the development of IL-4-producing cells was reduced in PD-L1(-/-) mice, demonstrating a role for PD-L1 in regulating Th cell differentiation. This inadequate Th2 response may explain the increased resistance of PD-L1(-/-) mice. Although no alterations in Th1/Th2 skewing were observed in PD-L2(-/-) mice, PD L2(-/-) mice exhibited a marked increase in L. mexicana-specific antibody production. Increased Leishmania-specific IgG production may suppress the healing response through FcgammaR ligation on macrophages. Taken together, our results demonstrate that PD-L1 and PD-L2 have distinct roles in regulating the immune response to L. mexicana. PMID- 16358364 TI - Toll-like receptor 9-mediated induction of the immunosuppressive pathway of tryptophan catabolism. AB - A series of recent studies, including an article in this issue of the European Journal of Immunology, have demonstrated that the administration of CpG-rich oligodeoxynucleotides (CpG-ODN) in experimental settings may lead to the activation of the immunosuppressive pathway of tryptophan catabolism, depending on several factors, including the route of CpG-ODN administration. These studies call attention to the need for a careful evaluation of the modalities of inclusion of CpG-ODN in vaccines for human use. At the same time, these studies may offer novel opportunities for use of CpG-ODN as immunosuppressive agents and may also lead to an improved understanding of the cellular events mediated by Toll-like receptor 9 signaling. PMID- 16358365 TI - Patient, disease, and therapy-related factors that influence discontinuation of disease-modifying antirheumatic drugs: a population-based incidence cohort of patients with rheumatoid arthritis. AB - OBJECTIVE: A major challenge in management of rheumatoid arthritis (RA) is prediction of longterm response to disease-modifying antirheumatic drug (DMARD) treatment. Our objective was to identify the predictors of DMARD discontinuation in an incidence cohort of patients with RA followed continuously from their incidence date. METHODS: Members of a population-based incidence cohort of Rochester, Minnesota, residents aged > or = 18 years diagnosed with RA (by 1987 American College of Rheumatology criteria) from January 1, 1955, to January 1, 1995, were followed longitudinally through their complete medical records until January 1, 2001. Detailed drug exposure data were collected on all DMARD and glucocorticoid regimens. Subjects were considered exposed to a DMARD if duration of use was > or = 30 days. Time to discontinuation of DMARD was estimated using survival analysis techniques. Andersen-Gill models with multiple events per patient were used to assess the influence of demographics, calendar time, comorbidities, disease characteristics [disease duration, rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), joint counts, radiographic changes, nodules, RA complications], and therapy characteristics (DMARD use, singly or in combination, glucocorticoid use, first or subsequent regimen, effect of previous therapy) on time from DMARD initiation to discontinuation. RESULTS: The study population comprised 345 DMARD-treated patients (73% female) with mean age of 53.1 years and mean followup 15.4 years. Median time taking any DMARD was 16.0 months for the first, and 17.9 months for all regimens. Methotrexate (MTX) had the longest time to discontinuation, with a median of 30.3 months without folate, and 61.7 months with folate supplementation. Among the various disease characteristics examined, only higher ESR at DMARD initiation was significantly associated with a shorter time taking DMARD [hazard ratio (HR) 1.05 per 10 mm/h increase, 95% CI 1.02, 1.08]. In multivariable Andersen-Gill models considering all DMARD regimens, hydroxychloroquine use (HR 0.77, 95% CI 0.64, 0.92) and MTX use (HR with folate 0.39, 95% CI 0.30, 0.51; HR without folate 0.51, 95% CI 0.39, 0.67) were significantly associated with longer time to DMARD discontinuation, whereas prior MTX use (HR 1.96, 95% CI 1.57, 2.45) was associated with shorter time to DMARD discontinuation, after adjusting for age, sex, calendar year, Charlson comorbidity index, disease duration, and ESR at DMARD initiation. Disease duration was negatively associated with time to DMARD discontinuation; each 10 year increase in disease duration corresponded to a 14% decrease in the risk of discontinuation (HR 0.86, 95% CI 0.75, 0.98). CONCLUSION: Longer RA disease duration does not appear to increase the risk of DMARD discontinuation. However, high disease activity (as assessed by ESR) is associated with a higher likelihood of discontinuing DMARD. MTX failure may identify a subgroup of patients who are less likely to respond to other DMARD and therefore could be considered as candidates for biological therapies. PMID- 16358366 TI - Etanercept in combination with sulfasalazine, hydroxychloroquine, or gold in the treatment of rheumatoid arthritis. AB - OBJECTIVE: To prospectively determine the efficacy and safety of etanercept in combination with sulfasalazine (SSZ), hydroxychloroquine (HCQ), and gold in the treatment of rheumatoid arthritis (RA). METHODS: A prospective open-label study enrolled 119 patients with RA who had active disease despite stable therapy with SSZ (n = 50), HCQ (n = 50), or intramuscular gold (n = 19). Primary efficacy endpoints consisted of American College of Rheumatology responses at 24 and 48 weeks. Safety was established at regularly scheduled visits. RESULTS: Patients in each etanercept combination showed significant improvement at both 24 and 48 weeks. Toxicity withdrawals by 48 weeks included gold (n = 1): proteinuria; HCQ (n = 5): septic wrist and bilateral pneumonia, rash, optic neuritis, breast cancer, squamous cancer of the tongue; and SSZ (n = 5): otitis media, elevated liver function indicators, pericarditis, rash, and gastroenteritis. The most common adverse events not requiring discontinuation from the study were injection site reactions (43% of patients) and upper respiratory type infections (34%). CONCLUSION: This study is the first to prospectively evaluate the safety of etanercept in combination with SSZ, HCQ, and gold in patients with RA. Etanercept in combination with SSZ, HCQ, or gold was efficacious and well tolerated, with a discontinuation rate of 9% (11/119) for adverse events at 48 weeks. PMID- 16358368 TI - What do osteoarthritis health outcome instruments measure? Impairment, activity limitation, or participation restriction? AB - OBJECTIVE: To explore whether commonly used osteoarthritis (OA) health outcome instruments (and items) are measuring single or multiple health outcomes using the International Classification of Functioning, Disability and Health (ICF) definitions. METHODS: Ten expert judges allocated 342 items from 13 instruments to one or more ICF construct, i.e., Impairment (I), Activity Limitation (A), and Participation Restriction (P). One-sample t tests were used to classify each item as measuring uniquely I, A, or P or some combination (i.e., IA, IP, AP, or IAP). RESULTS: Overall, 12 of the 13 instruments had items that measured a combination of outcome domains (i.e., IA, IP, AP, or IAP). Only the American Knee Society Score (AKS) had all items uniquely measuring either I or A. The instrument with the best representation of items for Impairment was the AKS, for Activity Limitation the WOMAC and Lequesne knee index, and for Participation Restriction the Disease Repercussion Profile. CONCLUSION: All the existing OA outcome instruments, except one, had some items that were assessing more than one health outcome. Use of these instruments may either mask true treatment effects or make an effect difficult to attribute if the content is unclear. We determined which instruments were the best for measuring each health outcome. To improve the assessment of health outcomes in OA, new instruments that uniquely measure the 3 ICF constructs should be developed and all 3 should be included in relevant studies. PMID- 16358369 TI - Study on the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell lines. AB - OBJECTIVE: Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes to clarify if hTRF1 mutation is one of the factors of the activation of telomerase. METHODS: hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification (TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells. RESULTS: hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as PsihTRF1-13, PsihTRF1-18, PsihTRF1-21 and PsihTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108-0.0749) than in normal mononuclear cells (0.0493, 0.0369 0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies. CONCLUSION: hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease. PMID- 16358370 TI - Determination of theophylline concentration in serum by chemiluminescent immunoassay. AB - OBJECTIVE: This study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum. METHODS: To measure the concentration of theophylline (n=122) and evaluate the assay. RESULTS: The linear range of the CLIA method was 0.51-40 mg/L (Y=1.02X+0.44, r=0.995). The intra and inter CV (coefficient variance) of CLIA were 3.20% and 3.57%, respectively. The average recovery rate was 102.3%. This method was free from interference by brilirubin (<200 micromol/L), hemoglobin (<10 g/L), and triglycerides (<15 mmol/L). CONCLUSION: This method is simple, convenient and precise for clinical pharmacokinetics study of theophylline. PMID- 16358371 TI - Segmental radiofrequency ablation of pulmonary vein ostia for patients with refractory paroxysmal atrial fibrillation using multi-slice spiral computed tomography guidance. AB - OBJECTIVE: To evaluate the safety and clinical efficacy of segmental radiofrequency ablation of pulmonary vein (PV) ostia for patients with refractory paroxysmal atrial fibrillation (AF) under multi-slice spiral computed tomography (MSCT) guidance before the procedure. METHODS: A series of 58 consecutive patients with refractory paroxysmal AF were enrolled to undergo segmental radiofrequency ablation of PV ostia. The 36 male and 22 female patients with mean age of (57.4+/-9.5) (32-79) years and no obvious organic heart disease. Before ablation, patients received MSCT to generate 3-dimensional image of the left atrium (LA) and proximal PVs. Patients then underwent segmental radiofrequency ablation of PV ostia using PV circular mapping catheter manipulated several times to ensure complete isolation between PVs and LA. RESULTS: No complications occurred during the procedure. One patient developed delayed cardiac tamponade, which was drained percutaneously. The mean follow-up time was (17.1+/-9.3) months. Forty-one patients (95%) experienced improved quality of life one month after the procedure. Thirty-six patients (83%) showed stable sinus rhythm, while 10 patients (23%) required additional anti-arrhythmic drugs. AF returned> or =1 time in 6 (14%) patients who underwent anti-arrhythmic drug therapy, but the number of episodes was less than that before the procedure. However, one patient experienced recurrent episodes of atrial flutter. CONCLUSION: It is safe and effective to perform segmental radiofrequency ablation of PV ostia for patients with refractory paroxysmal AF using MSCT guidance mappening. PMID- 16358372 TI - Validation of the use of foreign gas rebreathing method for non-invasive determination of cardiac output in heart disease patients. AB - OBJECTIVE: To compare a new device (Innocor) for non-invasive measurement of cardiac output (CO) by foreign gas rebreathing method with conventional techniques used in the measurements of cardiac function. METHODS: Cardiac outputs measured by Innocor (CO(RB)) were compared with CO obtained by echocardiography (CO(EC)), Swan-Ganz thermodilution (CO(TD)), and left ventricle radiography (CO(LVR)) in 34 patients subjected to cardiac catheterization. Values obtained from the four methods were analyzed by linear regression and paired values were compared by the method of Bland and Altman in SPSS. RESULTS: There was strong positive correlation (r=0.94) between Innocor cardiac output values and the corresponding values obtained by thermodilution and between CO(EC) and CO(LVR) values. Thermodilution appears to overestimate cardiac output when compared to the values obtained with Innocor by (0.66+/-0.22) L/min (P<0.0001). There was no correlation between data obtained by Innocor and the corresponding CO(EC) and CO(LVR) values. CONCLUSION: Innocor CO(RB) is an easy, safe and well established method for non-invasive measurement of cardiac output with good prospects for clinical application in heart disease patients. PMID- 16358373 TI - Prognostic significance of bcl-2 and p53 expression in colorectal carcinoma. AB - OBJECTIVE: This study was designed to detect the expression of bcl-2 and p53 proteins in colorectal carcinomas and to determine their association with the patient survival and stage of the diseases. METHODS: Immunohistochemistry method was used to detect the expression of bcl-2 and p53 proteins in 93 cases of colorectal carcinoma. The stain results were obtained by analyzing the clinic pathological characteristics of patients. RESULTS: Fifty-seven percent (53/93) of the colorectal carcinomas were bcl-2 protein positive. The positive rate of bcl-2 protein in lymph node involvement cases was lower (15/37) than the cases without node involvement (38/58, P<0.01). The positive rate of p53 protein was 43% (40/93) in colon-rectum carcinomas. No significant correlation was observed between p53 protein expression and clinic-pathological manifestations (P>0.05) but the survival was significantly worse (P=0.0001) in the p53 protein positive cases. Neither bcl-2 nor p53 alone was correlated with stage of the disease. When combined bcl-2/p53 status was analyzed, a group with bcl-2(+) and p53(-) had the best prognosis. This group was significantly associated with earlier Dukes' stages (P=0.1763). In multivariate Cox regression analysis, lymph node involvement and p53 protein expression were two independent factors correlated with survival time. CONCLUSION: The expression of bcl-2 and p53 represent biological characteristics of colorectal carcinomas. Assessment of both bcl-2 and p53 status may be valuable in predicting the prognosis of patients. PMID- 16358374 TI - Evaluation of ST13 gene expression in colorectal cancer patients. AB - We identified a novel gene ST13 from a subtractive cDNA library of normal intestinal mucosa in 1993, more studies showed that ST13 was a co-chaperone of Hsp70s. Recently we detected the ST13 gene expression in tumor tissue and adjacent normal tissue of the same colorectal cancer patient and investigated if the ST13 gene expression might have any prognostic value. Analysis was performed at molecular level by reverse transcription-PCR using real-time detection method. We measured two genes simultaneously, ST13 as the target gene and glyceraldehydes 3-phosphate dehydrogenase as a reference gene, in primary colorectal tumor specimens and tumor-adjacent normal mucosa specimens from 50 colorectal cancer patients. The expression levels of the ST13 gene were significantly decreased in primary tumors compared with adjacent mucosa (P<0.05). But there were no significant differences in the expression of ST13 as compared with different Dukes' stage, tumor differentiation grade, invasion depth, lymph node metastasis and disease-specific survival. PMID- 16358375 TI - Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes. AB - OBJECTIVE: This study is aimed at developing a simple and easy way to generate dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) in vitro. METHODS: PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods (collecting the attached cell and removing the suspension cell). DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor (rhGM CSF) and 20 ng/ml recombinant human interleukin-4 (rhIL-4) in vitro. On the sixth day, TNF-alpha (TNFalpha) 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions (MLR) were used to characterize DC function after TNFalpha or lipopolysaccharide (LPS) stimulations for 24 h. RESULTS: After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CD1a, CD80 and CD86, features of DCs. TNFalpha treatment induced DCs maturation and up regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity. CONCLUSION: This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs. PMID- 16358376 TI - Clinical features of chronic hepatitis B patients with YMDD mutation after lamivudine therapy. AB - OBJECTIVE: To study the clinical features of chronic hepatitis B (CHB) patients with tyrosine-methionine-aspartate-aspartate (YMDD) mutation after lamivudine therapy. METHODS: This investigation was a retrospective study of 63 CHB patients with YMDD mutation during lamivudine therapy. Clinical data, including period and types of YMDD mutation; hepatitis B virus (HBV) DNA levels and alanine aminotransferase (ALT) levels before and after YMDD mutation were measured. YMDD mutation in the HBV DNA polymerase gene was determined using polymerase chain reaction (PCR) and direct sequencing. HBV DNA quantification was determined using real-time PCR. Relevant serum markers of HBV were measured. The follow-up period was 12 months after YMDD mutation. RESULTS: YMDD mutation occurred 7-44 months (median, 21.5 months) after the start of lamivudine therapy. The majority of the cases (42/63, 66.6%) had YMDD mutants detected between 12 and 24 months. Four types of YMDD mutation were observed in this study, rtL180M/M204V mutation was the predominant type (26/63, 41.3%). A proportion of patients (16/63, 25.4%; 12/63, 19.1%) had higher HBV DNA levels and ALT levels (after mutation vs before mutation), respectively. CONCLUSION: The majority of patients with YMDD mutants had similar or lower HBV DNA levels and ALT levels compared with baseline values. This subset of patients might have benefited from the continued lamivudine therapy. The patients with increased ALT and HBV DNA levels (breakthrough hepatitis) should benefit from the addition of a newer nucleotide analogue (e.g. adefovir). PMID- 16358377 TI - Infusion of nonmyeloablative bone marrow alleviates acute rejection reaction in liver allotransplantation. AB - OBJECTIVE: To study the effect and implication of nonmyeloablative donor specific bone marrow (DSBM) infusion on the immunoreaction of liver allotransplantation. METHODS: Orthotopic liver transplantation model was used in this study. Groups were set as follows: Group I, syngeneic control (Wistar-to-Wistar); Group II, acute rejection (SD-to-Wistar); Group III, acute rejection treated with cyclosporine A (CsA) by intramuscular injection (SD-to-Wistar+CsA); Group IV, bone marrow infusion at 7 d pretransplantation followed by short-term CsA treatment (SD-to-Wistar+DSBM); Another group of short-term CsA treatment preoperatively without bone marrow infusion was also set as control. General characteristics and survival time were observed. Histological grades of rejection were determined by pathological examination. IL-2 and IFN-gamma level in peripheral blood and donor liver were detected respectively by Enzyme-Linked Immuno-Sorbent Assay (ELISA) and Western blot. Chimerism of donor cells was measured by PCR for a male-specific marker (Y-chromosome-specific sequence, Sry). RESULTS: No signs of rejection were found in Group I. Acute rejection occurred in both Group II and the short-term CsA treated group. All the recipients died at (9 15) d posttransplantation with a median survival time of (10.7+/-0.5) d and (11.2+/-2.4) d, respectively. Only mild rejection could be seen in Group III. In Group IV, 4 out of 6 recipients had long-term survival (>100 d), the histological grade of rejection was significantly lower than that of Group II, so did the expression level of IL-2 and IFN-gamma in both peripheral blood and grafted liver. Y-chromosome-specific sequence (Sry) of male SD rats could be detected in the bone marrow, spleen and thymus of female recipients at 15 d after bone marrow infusion. CONCLUSION: Mild preconditioning nonmyeloablative donor specific bone marrow infusion can enhance chimerism formation in recipients, alleviate the rejection of liver allotransplantation and prolong survival of liver allotransplantation. PMID- 16358378 TI - Comparison of low-molecular-weight-heparin and unfractionated heparin for acute PTE. AB - OBJECTIVE: Acute pulmonary thromboembolism (PTE) is a serious high mortality pulmonary vascular disease whose effective treatment decreases morbidity and mortality. To determine if low-molecular-weight-heparin (LMWH) is clinically as efficient and safe as unfractionated heparin (UH) in patients with diagnosis of acute non-massive PTE, our study compares the efficacy, adverse effects and costs of LMWH and UH. METHODS: One hundred and fourteen patients with non-massive acute PTE were randomly divided into LMWH (nadroparin calcium) and UH groups. Oxygenation index, D-dimer, fibrinogen (FG), lung ventilation/perfusion (V/Q) scan and computed tomography pulmonary angiography (CTPA) were observed before anticoagulation and on day 14 after anticoagulation. RESULTS: In both groups, the ABG (arterial blood gas) analysis showed PaO(2) and PaCO(2) were elevated, P(A a)O(2) was decreased and oxygenation index (PaO(2)/FIO(2)) was elevated, D-dimer and fibrinogen were decreased, lung V/Q and CTPA showed embolized segments reduced (P<0.05). Hemorrhage and thrombocytopenia occurred in 3.5% of the LMWH group. Hemorrhage occurred in 5.3% and thrombocytopenia occurred in 7.0% of the UH group. The average cost in the LMWH group was RMB 1218.60 Yuan and RMB 1541.40 Yuan in the UH group. CONCLUSION: LMWH and UH are equally effective for treatment of non-massive acute PTE, but LMWH may have a lower prevalence of complications and is less expensive. PMID- 16358379 TI - Investigation of iodine concentration in salt, water and soil along the coast of Zhejiang, China. AB - OBJECTIVE: We aim to describe the environment iodine concentration in salt, water and soil along Zhejiang Province coast in the China foreland. It will be helpful for us to judge whether this area is insufficient in iodine and universal iodized salt is necessary or not. METHODS: We collected iodized salt samples, drinking water samples (tap water in the towns, and well water or spring water in the villages), water samples from different sources (ditches, lakes, rivers) and soil samples through random sampling in June, 2005. Salt, water and soil iodine was detected by arsenic-cerium redox method. Statistical analysis was expressed as mean+/-SEM by Windows SPSS 13.0. RESULTS: (1) The iodine concentration in salt was 27.9+/-4.33 mg/kg (n=108). (2) Seventy-five water samples were collected. The water iodine value was 0.6-84.8 microg/L (mean of 11.66 mug/L). The watershed along the Qiantang River has significantly higher iodine content than the water in Lin'an in mountain area (P<0.01). The iodine content and mean iodine content of tap water, well or spring water and natural water sources were 4.30+/-2.43 microg/L (n=34), 23.59+/-27.74 microg/L (n=19) and 12.72+/-10.72 microg/L (n=22) respectively. This indicated that among environmental water sources, the ditch iodine content was the highest with river water iodine being the lowest (P<0.01). (3) Soil iodine value was 0.11-2.93 mg/kg (mean of 1.32 mg/kg). Though there was no statistical difference of soil iodine in different districts (P=0.131), soil iodine content correlated positively with water iodine content. CONCLUSION: Iodine concentration in salt accords with national policy of adding iodine in salt. Foreland has more iodine in water than mountain area. The data reflected that water and soil iodine in foreland area was not high, which suggests universal iodized salt should be necessary. Environment iodine has relatively close association with pollution. PMID- 16358380 TI - Treatment and follow-up of children with transient congenital hypothyroidism. AB - OBJECTIVE: To study the clinical therapy and prognosis in children with transient congenital hypothyroidism (CH). METHODS: Fifty-seven children with CH diagnosed after neonatal screening were treated with low-dosage levothyroxine (L-T4). Follow-up evaluation included the determination of TT3, TT4 and TSH serum levels and the assessment of thyroid gland morphology, bone age, growth development and development quotients (DQ). A full check-up was performed at age 2, when the affected children first discontinued the L-T4 treatment for 1 month, and one year later. Development quotients were compared with a control group of 29 healthy peers. RESULTS: The initial L-T4 dosage administered was 3.21-5.81 microg/(kg.d) with an average of (16.25+/-3.87) microg/d. Mean duration of therapy was (28.09+/ 9.56) months. No significant difference was found between study group and control group in the DQ test (average score (106.58+/-14.40) vs (102.4+/-8.6), P>0.05) and 96.49% of the CH children achieved a test score above 85. Bone age, 99mTc scans and ultrasonographic findings were all normal, and evaluation of physical development was normal too, as were the serum levels of TT3, TT4 and TSH after one year of follow-up. CONCLUSION: A L-T4 dosage of 3.21-5.81 microg/(kg.d) was found sufficient for the treatment of transient CH. The treated children showed satisfactory overall mental and physical development at age 2. So it is possible for CH children to stop taking medicine if their laboratory findings and physical development are all normal after regular treatment and 2-3 years of follow-up. PMID- 16358381 TI - A novel technique of three-dimensional reconstruction segmentation and analysis for sliced images of biological tissues. AB - A novel technique of three-dimensional (3D) reconstruction, segmentation, display and analysis of series slices of images including microscopic wide field optical sectioning by deconvolution method, cryo-electron microscope slices by Fourier Bessel synthesis and electron tomography (ET), and a series of computed tomography (CT) was developed to perform simultaneous measurement on the structure and function of biomedical samples. The paper presents the 3D reconstruction segmentation display and analysis results of pollen spore, chaperonin, virus, head, cervical bone, tibia and carpus. At the same time, it also puts forward some potential applications of the new technique in the biomedical realm. PMID- 16358382 TI - Study on EEG power and coherence in patients with mild cognitive impairment during working memory task. AB - To investigate the features of electroencephalography (EEG) power and coherence at rest and during a working memory task of patients with mild cognitive impairment (MCI). Thirty-five patients (17 males, 18 females; 52-71 years old) and 34 sex- and age-matched controls (17 males, 17 females; 51-63 years old) were recruited in the present study. Mini-Mental State Examination (MMSE) of 35 patients with MCI and 34 normal controls revealed that the scores of MCI patients did not differ significantly from those of normal controls (P>0.05). Then, EEGs at rest and during working memory task with three levels of working memory load were recorded. The EEG power was computed over 10 channels: right and left frontal (F3, F4), central (C3, C4), parietal (P3, P4), temporal (T5, T6) and occipital (O1, O2); inter-hemispheric coherences were computed from five electrode pairs of F3-F4, C3-C4, P3-P4, T5-T6 and O1-O2 for delta (1.0-3.5 Hz), theta (4.0-7.5 Hz), alpha-1 (8.0-10.0 Hz), alpha-2 (10.5 -13.0 Hz), beta-1 (13.5 18.0 Hz) and beta-2 (18.5-30.0 Hz) frequency bands. All values of the EEG power of MCI patients were found to be higher than those of normal controls at rest and during working memory tasks. Furthermore, the values of EEG power in the theta, alpha-1, alpha-2 and beta-1 bands of patients with MCI were significantly high (P<0.05) in comparison with those of normal controls. Correlation analysis indicated a significant negative correlation between the EEG powers and MMSE scores. In addition, during working memory tasks, the EEG coherences in all bands were significantly higher in the MCI group in comparison with those in the control group (P<0.05). However, there was no significant difference in EEG coherences between two groups at rest. These findings comprise evidence that MCI patients have higher EEG power at rest, and higher EEG power and coherence during working conditions. It suggests that MCI may be associated with compensatory processes at rest and during working memory tasks. Moreover, failure of normal cortical connections may be exist in MCI patients. PMID- 16358383 TI - Sensory exotropia subsequent to senile cataract. AB - To evaluate the phacoemulsification and intraocular lens implantation in patients with sensory exotropia subsequent to senile cataract. The authors prospectively studied the role of phacoemulsification and intraocular lens implantation on 25 patients by observing visual acuity, ocular alignment, binocular vision and diplopia pre-, 1 month post- and 3 months post-operation. The patients underwent follow-up for three months. Postoperatively, one patient had a corrected visual acuity of 20/50, and 24 patients had 20/40 or better. The ocular alignment, binocular vision and diplopia were resolved spontaneously. Phacoemulsification and intraocular lens implantation performed together is effective on sensory exotropia subsequent to senile cataract. PMID- 16358385 TI - Bibliography. Current world literature. Ocular manifestations of systemic disease part I. PMID- 16358384 TI - Report--21st century medical genetic and genomic medicine in China. PMID- 16358386 TI - Healthy eating. Salad to squash. Vary your veggies. PMID- 16358387 TI - Does sex matter? Diabetes, heart disease, and gender. PMID- 16358388 TI - Back to basics. Blood pressure. PMID- 16358389 TI - Diabetes care when you're sick. Flu season is upon us. Here's what to do when you're feeling under the weather. PMID- 16358390 TI - Warm chili for a cold day. PMID- 16358391 TI - Keeping off lost weight. For more than a decade, the National Weight Control Registry has been tracking people who have lost weight and kept it off. PMID- 16358392 TI - Research profile. Hormonal birth control. A gestational diabetes risk? Assiamira Ferrara, M.D., Ph.D. PMID- 16358393 TI - Quantitative assessment of regional myocardial function with MR-tagging in a multi-center study: interobserver and intraobserver agreement of fast strain analysis with Harmonic Phase (HARP) MRI. AB - PURPOSE: To assess the reproducibility of Harmonic Phase (HARP) analysis of myocardial MR tagged images acquired in the Multi-Center Study of Atherosclerosis (MESA). METHODS: Using the HARP method, three independent observers performed two separate quantitative strain analyses of myocardial cine MR-tagging images blindly in 24 participants. The images were obtained in four different centers and analyzed at a single core lab. Each study comprised 3 short-axis slices subdivided in 12 segments (24 x 3 x 12 = 864 segments), each with three layers. Normal strains (circumferential [Ecc] and radial [Err]), principal strains (Lambda1, Lambda2), and the angle alpha (between Ecc-Lambda2) were calculated. Intraclass correlation coefficient (R) for peak systolic strains, and all pooled systolic and diastolic strain data were used to determine inter- and intraobserver agreement. Two observers also visually graded study quality. R values were related to the image quality in different myocardial regions and layers. RESULTS: Overall, HARP yielded an excellent inter- and intraobserver agreement for peak systolic strain data (for Ecc, R = 0.84 and 0.89, respectively) and all systolic pooled data (for Ecc, interobserver R = 0.82, intraobserver R = 0.69-0.76). Both inter and intraobserver agreement were lower for diastolic pooled data (R = 0.69 and 0.58-0.62, respectively). There was a direct relationship between image quality and performance of the HARP analysis, with increasing inter- and intraobserver R values in studies with longer tag persistence. Both inter- and intraobserver agreement were better in the anterior and septal myocardial regions, and in the midwall layer. The intraobserver agreement was similar among the three observers. CONCLUSION: Employing the HARP method for quantitative strain analysis of myocardial MR tagged images provides a high inter- and intraobserver agreement. These good results are obtained in case of good to excellent MR image quality. PMID- 16358394 TI - Dietary and physical activity correlates of long-term weight loss. AB - Covariations in body mass index (BMI), physical activity, macronutrient intake, and the frequency of consumption of specific foods were examined among 82 men and 75 women participating in a behavioral weight loss program over a period of 18 months. Results of repeated measures analyses of covariance showed that BMI change was inversely related to change in physical activity and change in frequency of vegetable consumption. BMI change was positively related to change in calorie intake from fat and change in frequency of consumption of beef, hot dogs, and sweets. Change in fat calories predicted BMI change better than change in total calories. In addition, change in the frequency of consumption of specific foods accounted for a larger percentage of the variance in BMI change than did change in macronutrients (10.4% vs. 5.2%). No differences were found between predictors of weight loss vs. weight maintenance. PMID- 16358395 TI - Metabolic differences in response to a high-fat vs. a high-carbohydrate diet. AB - Energy expenditure was measured in a group of 7 subjects who received two isocaloric isonitrogenous diets for a period of 9-21 days with a 4-10-day break between diets. Diet 1 was a high-fat diet ( 83.5 +/- 3.6% of total energy). Diet 2 was a high carbohydrate diet ( 83.1 +/- 3.7% of total energy). Resting and postprandial resting metabolic rate were measured by open circuit indirect calorimetry 2-4 times during each metabolic period. Total energy expenditure (TEE) was measured by the doubly labeled water method over an 8-13-day period. The respiratory quotient was measured 2-4 hours after a meal during each metabolic period for the calculation of total energy expenditure by the doubly labeled water method. Levels of total T3 (TT3), T3 uptake, free thyroid index and T4 were measured at the end of each metabolic period. No significant changes in resting metabolic rate (RMR) were apparent on the two diets (1567 +/- 426 kcal/d high-fat diet and 1503 +/- 412 kcal/d high-carbohydrate diet n=7, p<0.15). Total energy expenditure measured in 5 subjects was significantly higher during the high-carbohydrate phase of the diet (2443 +/- 422 vs. 2078 +/- 482 kcal/d p<0.05). Activity estimated from TEE/RMR was greater on the high-carbohydrate diet but only approached statistical significance (p<0.06). Total T3 was significantly lower and free thyroid index and T3 uptake were significantly higher at the end of the high fat diet in comparison to the high-carbohydrate diet. These data suggest that individual tolerance to a high-fat diet varies considerably and may significantly lower TEE by changing levels of physical activity. The explanation for changes in thyroid hor. mone levels independent of changes in metabolic rate remains unclear. PMID- 16358396 TI - Binge eating behavior in patients with eating disorders. AB - The purpose of this study was to determine whether the objectively observed binge eating behavior of obese subjects meeting the proposed DSM-IV criteria for binge eating disorder would be similar to that observed in patients with bulimia nervosa. Non-obese patients with bulimia nervosa (BN), obese subjects with binge eating disorder (BED), obese and non-obese women without eating disorders were each instructed to binge eat single- and multiple-item meals. In the multiple item meal, the obese subjects with BED ate significantly more (1515 kcal) than obese subjects without BED (1115 kcal), but they ate less than the normal-weight bulimic patients (2680 kcal). The non-obese controls ate amounts similar to the obese non-binge-eating-disordered group (1093 and 1115.2 kcal, respectively). In the single-item meal, consisting of ice cream, patients with BN ate significantly more than any other group (1307 kcal), while obese subjects with or without binge eating disorder ate significantly more (762 kcal) than non-obese controls (308 kcal). This study has demonstrated that although both BN and BED are characterized by recurrent episodes of binge eating, quantitatively there appear to be differences between the eating disturbances in the two disorders. Because single- and multiple-item meals differ in external cues, these results also suggest that the obese subjects with BED may be disinhibited by external cues, while obese subjects without BED may be inhibited by external cues. PMID- 16358397 TI - The response to exercise with constant energy intake in identical twins. AB - Seven pairs of young adult male identical twins completed a negative energy balance protocol during which they exercised on cycle ergometers twice a day, 9 out of 10 days, over a period of 93 days while being kept on a constant daily energy and nutrient intake. The total energy deficit caused by exercise above the estimated energy cost of body weight maintenance reached 244 +/- 9.8 MJ (Mean +/- SEM). Baseline energy intake was estimated over a period of 17 days preceding the negative energy balance protocol. Mean body weight loss was 5.0 kg (SEM = 0.6) (p < 0.001) and it was entirely accounted for by the loss of fat mass (p < 0.001). Fat-free mass was unchanged. Body energy losses reached 191 MJ (SEM = 24) (p < 0.001) which represented about 78% of the estimated energy deficit. Subcutaneous fat loss was slightly more pronounced on the trunk than on the limbs as estimated from skinfolds, circumferences, and computed tomograply (CT). The reduction in CT assessed abdominal visceral fat was quite striking, from 81 cm2 (SEM = 5) to 52 cm2 (SEM = 6) (p < 0.001). At the same submaximal power output level, subjects oxidized more lipids than carbohydrates after the program as indicated by the changes in the respiratory exchange ratio (p < or = 0.05). Intrapair resemblance was observed for the changes in body weight (p < 0.05), fat mass (P < 0.01), percent fat (p < 0.01), body energy content (p < 0.01), sum of 10 skinfolds (p < 0.01), abdominal visceral fat (p < 0.01), fasting plasma triglycerides (p < 0.05) and cholesterol (p < 0.05), maximal oxygen uptake (p < 0.05), and respiratory exchange ratio during submaximal work (p < 0.01). We conclude that even though there were large individual differences in response to the negative energy balance and exercise protocol, subjects with the same genotype were more alike in responses than subjects with different genotypes particularly for body fat, body energy, and abdominal visceral fat changes. High lipid oxidizers and low lipid oxidizers during submaximal exercise were also seen despite the fact that all subjects had experienced the same exercise and nutritional conditions for about three months. PMID- 16358399 TI - Child and parent psychological problems influence child weight control. AB - This study used path analytic methods to assess the influence of child psychopathology measured using the Child Behavior Checklist and parent psychiatric symptoms measured using the Cornell Medical Index on changes in child percent overweight in obese 8- to 11-year-old children over 2 years (6 months of treatment, 18 months of follow-up) provided family-based behavioral intervention. Path analysis for the 0-6 month change showed the influence of mother and father psychiatric symptoms and child age on child anxiety/depression scores that in turn influenced child percent overweight change during treatment, accounting for 41% of the shared variance. The path analysis for follow-up change showed mother and father psychiatric symptoms influenced child social problems at the first stage, and child social problems influenced child percent overweight change, accounting for 68.5% of the shared variance. These results provide the first demonstration that both parent and child problems may influence the short- and long-term success of obese children who participate in family-based behavioral treatment programs. PMID- 16358398 TI - The male obese Wistar diabetic fatty rat is a new model of extreme insulin resistance. AB - The male obese Wistar Diabetic Fatty (WDF) rat is a genetic model of obesity and non-insulin dependent diabetes (NIDDM). The obese Zucker rat shares the same gene for obesity on a different genetic background but is not diabetic. This study evaluated the degree of insulin resistance in both obese strains by examining the binding and post binding effects of muscle insulin receptors in obese rats exhibiting hyperinsulinemia and/or hyperglycemia. Insulin receptor binding and affinity and tyrosine kinase activity were measured in skeletal muscle from male WDF fa/fa (obese) and Fa/? (lean) and Zucker fa/fa (obese) and Fa/Fa (homozygous lean) rats. Rats were fed a high sucrose (68% of total Kcal) or Purina stock diet for 14 weeks. At 27 weeks of age, adipose depots were removed for adipose cellularity analysis and the biceps femoris muscle was removed for measurement of insulin binding and insulin-stimulated receptor kinase activity. Plasma glucose (13.9 vs. 8.4 mM) and insulin levels (14,754 vs. 7440 pmol/L) were significantly higher in WDF obese than in Zucker obese rats. Insulin receptor number and affinity and TK activity were unaffected by diet. Insulin receptor number was significantly reduced in obese WDF rats ( 2.778 +/- 0.617 pmol/mg protein), compared to obese Zucker rats (4.441 +/- 0.913 pmol/mg potein). Both obese strains exhibited down regulation of the insulin receptor compared to their lean controls. Maximal tyrosine kinase (TK) activity was significantly reduced in obese WDF rats (505 +/- 82 fmol/min/mg protein) compared to obese Zucker rats (1907 +/- 610 fmol/min/mg protein). Only obese WDF rats displayed a decrease in TK activity per receptor. These observations establish the obese WDF rat as an excellent model for exploring mechanisms of extreme insulin resistance, particularly post-receptor tyrosine kinase-associated defects, in non-insulin dependent diabetes. PMID- 16358400 TI - Associations of body mass and body fat distribution with parity among African American and Caucasian women: The CARDIA Study. AB - Associations of parity with body fat and its distribution are poorly understood; therefore, we examined the relationships between parity and obesity in young adult women. Body mass index (BMI), skin-folds, and waist-hip ratio were compared in 1452 African-American and 1268 Caucasian nonpregnant women aged 18 to 30, adjusting for age (where no age-parity interactions were present), education, physical activity (assessed by questionnaire) and fitness (assessed by graded exercise test), dietary fat intake, alcohol and smoking. Adjusted mean BMI was significantly higher in African-American women aged 25-30 years with three or more children (28.5 kg/m2) than in those with two (27.0 kg/m2), one (26.2 kg/m2), or no children (26.3 kg/m2). Similar trends were found in Caucasians (BMI = 23.3, 23.4, 23.7, 25.0 kg/m2 for parity = 0, 1, 2, > or = 3, respectively), but the mean BMI was significantly higher in African Americans in each parity group. The association between BMI and parity was not present among women 18-24 years of age. Skinfolds were directly associated with parity in African Americans only. Waist-hip ratios were generally lower among nulliparous than parous women in both ethnic groups; race differences were present only among nulliparas. In conclusion, parity was associated with BMI in women aged 25 to 30 years but did not explain ethnicity-related differences in body mass. PMID- 16358401 TI - Vagal-central nervous system interactions modulate the feeding response to peripheral enterostatin. AB - Enterostatin selectively inhibits the intake of dietary fat after both peripheral and central administration. We have investigated the role of the hepatic vagus nerve in modulating the peripheral response to enterostatin in Sprague-Dawley rats adapted to a high fat (HF) diet. Intraperitoneal (ip) enterostatin reduced intake of HF diet after overnight starvation. This response was abolished by selective vagal hepatic branch transection. Immunohistochemical techniques were used to identify the location of Fos protein in brain nuclei after ip enterostatin. Fos protein was evident in the nucleus tractus solitarius (NTS), parabrachial, paraventricular and supraoptic nuclei. The pattern of expression of Fos-like immunoreactivity differed from that induced by the lipoprivic agent beta mercaptoacetate. Transection of the hepatic vagus blocked the central Fos responses to ip enterostatin. We conclude that afferent hepatic vagal nerve activity is required for the feeding response to peripheral enterostatin. PMID- 16358402 TI - Use of glycated hemoglobin to assess Glycemic control in Wistar diabetic fatty rats and Zucker fatty rats. AB - The Wistar Diabetic Fatty rat (WDF fafa) is a congenic strain of the Wistar Kyoto rat. Studies using blood glucose reveal that only fatty male (not female) WDF rats spontaneously develop hyperglycemia when fed a stock diet. Blood glucose values have not provided consistent results for evaluation of glycemic status in fatty male WDF rats. Zucker fatty (fafa) rats, while sharing the fa gene and the development of hyperinsulinemia and hyperlipemia, do not spontaneously become hyperglycemic. In order to examine strain differences and the effects of age on long-term average glycemic status in WDF and Zucker rats, glycated hemoglobin (GHb) was analyzed. Glycated hemoglobin was measured in male lean and obese WDF and Zucker rats at 2, 3, 6, and 12 months of age. Nonfasted plasma glucose was measured in male lean and obese WDF rats at 2, 3, 6, and 12 months of age and in lean and obese Zucker rats at 3, 6, and 12 months of age. Plasma insulin was measured in lean and obese WDF and Zucker rats at 3, 6, and 12 months of age. Obese WDF rats had significantly elevated GHb compared to lean controls at 3, 6, and 12 months of age. Glycated hemoglobin was substantially above the normal range (3.8-6.5%) at 3 months of age (14.1%). Glycated hemoglobin significantly declined in the obese WDF rats between 6 and 12 months of age. Nonfasted plasma glucose was significantly elevated in the obese WDF rats at 3 months (14.1 +/- 2.1 mM/L) and 6 months of age (16.2 +/- 2.3 mM/L) compared to lean controls. At 12 months of age there was no difference in plasma glucose between obese and lean WDF rats. Obese and lean Zucker rats had similar levels of GHb and plasma glucose at all ages. In conclusion, GHb provides more integrated data for classifying disease status of WDF rats and evaluation of potential long-term complications associated with hyperglycemia. PMID- 16358403 TI - Chronic calorie restriction alters glycogen metabolism in rhesus monkeys. AB - Chronic caloric restriction (CR) prevents the development of obesity and maintains health, slows aging processes, and prevents or substantially delays the development of non-insulin-dependent diabetes. Because changes in energy metabolism could be involved in all of these positive effects of CR, we examined glycogen synthase (GS) and glycogen phosphorylase (GP) activities and glucose 6 phosphate (G6P) and glycogen concentrations in skeletal muscle samples before and during a euglycemic hyperinsulinemic clamp in 6 older aged monkeys in which CR had been continued for 10.4 +/- 2.1 years. Basal GS activity (fractional velocity and independent) was significantly higher in the CR monkeys than has been previously shown in normal, hyperinsulinemic and diabetic monkeys. The normal effect of insulin to activate GS was absent in the CR group due to the paradoxical finding in some of these monkeys of a reduction in GS activity by insulin. Insulin also had the unexpected effect of increasing the independent activity of GP above basal activity (p<0.05). There was an inverse relationship between the change (insulin-stimulated minus basal) in GS fractional velocity and GP activity ratio (r=-0.91, p<0.005). The basal independent activities of GS and GP were also inversely correlated (r=-0.79, p<0.05). The insulin-stimulated concentration of G6P tended to be higher than the basal concentration (p<0.06) and was significantly higher than that previously shown in normal monkeys (p<0.05). We suggest that long-term calorie restriction (1) results in alterations in glycogen metabolism that may be important to the anti-diabetogenic and antiaging effects of CR and (2) unmasks early defects which may indicate the likelihood of ultimately developing obesity and diabetes. PMID- 16358404 TI - Weight-loss strategies of young adults: exercise versus dieting. AB - This cross-sectional survey study examined weight control practices of 2,092 male and 1,748 female feshman students aged 18 years. For women, the desire for weight loss was observed at all levels of body weight. Most women (79%) wished to weigh less than their current weight, and 23% were following a restricted-calorie diet. In contrast, more men wished to gain (46%) rather than lose weight (32%), and only 3% were dieting at the time of the study. A comparison of weight control practices of men and women who wished to lose weight showed that women restricted calories far more frequently than did men. In contrast, although men were more likely to use exercise for weight control than were women, the differences between the sexes were small. A combined analysis of dieting and exercise behaviors showed that while men relied almost exclusively on intensive exercise as opposed to dieting, women employed exercise as well as dieting for the purpose of weight control. PMID- 16358405 TI - Physical activity and long-term maintenance of weight loss. AB - To examine the effect of exercise on the long-term maintenance of weight loss, two types of literature were reviewed--correlational studies of predictors of long-term weight loss, and randomized trials comparing diet, exercise, and the combination of diet plus exercise. Both literatures were striking in the consistency with which activity emerged as a determinant of long-term maintenance of weight loss. The benefits of exercise for long-term weight maintenance were observed with different types of populations, diets, and exercise interventions. Several possible explanations for these positive effects of diet plus exercise are presented, and suggestions made for future research on ways to maximize the benefit of this approach to weight control. Since adherence to exercise may ultimately prove to be the cornerstone for long-term weight maintenance, studying ways to improve exercise adherence is recommended. PMID- 16358406 TI - Regulation of microtubules by Rho GTPases in migrating cells. AB - Microtubules (MTs) contribute to cell polarization and migration, but the molecular mechanism involved are unknown. We have explored signalling pathways that generate specific changes in MTs arrays in wounded monolayers of fibroblasts. In earlier work, we found that Rho GTPase and its effector mDia, stimulate selective MT stabilization in the lamella, whereas Cdc42 and the MT motor protein dynein regulate MT organizing centre (MTOC) reorientation towards the leading edge. We have now found that the MT tip proteins EB1 and adenomatous polyposis coli protein (APC) function with mDia to stabilize MTs and interact directly with mDia. EB1, APC and mDia localize to the ends of stabilized MTs suggesting that they may contribute to capping of these MTs. Models of MTOC reorientation suggest that the MTOC moves in front of the nucleus by dynein pulling on MTs. In contrast, we find by directly imaging MTOC reorientation that the nucleus moves rearward while the MTOC remains stationary. Rearward nuclear movement is coupled to retrograde actin-myosin flow and is regulated by Cdc42 and its effector myotonic dystrophy kinase-related Cdc42-binding kinase. Dynein is not involved in nuclear movement, but is essential to maintain the MTOC at the cell centroid. These results show that there are two Cdc42 pathways that regulate MTOC reorientation. PMID- 16358407 TI - Interaction of cadherin with the actin cytoskeleton. AB - Cadherins regulate cell-cell adhesion throughout embryonic development and in the adult organism, and defects in cadherin expression and function are characteristic of many disease states including cancer. Although extracellular binding between cadherins specifies adhesion between cells, the strength of the interaction is thought to be regulated by cadherin clustering through reorganization of the actin cytoskeleton. Protein-protein interactions have been described that could link cadherins either directly or indirectly to the actin cytoskeleton. Here, we describe these protein interactions, and examine critically the evidence that they link cadherins to the actin cytoskeleton. PMID- 16358408 TI - Rho GTPase-formin pairs in cytoskeletal remodelling. AB - Diaphanous-related formins (Drfs) are members of a conserved formin family of actin-nucleating proteins and are thought to act as Rho GTPase effectors in signal transduction pathways that govern gene expression, cytoskeletal remodelling and cell division. In vitro evidence suggests that the three mammalian Drf proteins--mDia1, mDia2 and mDia3-have distinct GTPase-binding specificities. However, much of our current understanding of GTPase-Drf partnerships in mammalian cell signalling is based on expression studies using Drfs missing their unique GTPase-binding domains. We have employed fluorescence resonance energy transfer (FRET) and gene targeting approaches to identify the function of different GTPase-formin pairs in cell signalling. These studies have allowed us to uncover new roles for Drf proteins in cytoskeletal remodelling and novel regulatory mechanisms whereby GTPases influence formin function. Our genetic experiments strongly suggest that Drfs cooperate with other GTPase effector proteins, including the gene product of the Wiskott-Aldrich syndrome gene, WASP, during the regulation of cell proliferation. Further, the Drf gene knockout experiments indicate that this family of formins has a role in cancer pathophysiology. PMID- 16358409 TI - Survival of rat or mouse ventral mesencephalon neurons after cotransplantation with rat sertoli cells in the mouse striatum. AB - Transplanting cells across species (xenotransplantation) for the treatment of Parkinson's disease has been considered an option to alleviate ethical concerns and shortage of tissues. However, using this approach leads to decreased cell survival; the xenografted cells are often rejected. Sertoli cells (SCs) are testis-derived cells that provide immunological protection to developing germ cells and can enhance survival of both allografted and xenografted cells. It is not clear whether these cells will maintain their immunosuppressive support of cografted cells if they are transplanted across species. In this study, we investigated the immune modulatory capacity of SCs and the feasibility of xenografting these cells alone or with allografted and xenografted neural tissue. Transplanting xenografts of rat SCs into the mouse striatum with either rat or mouse ventral mesencephalon prevented astrocytic infiltration of the graft site, although all transplants showed activated microglia within the core of the graft. Surviving tyrosine hydroxylase-positive neurons were observed in all conditions, but the size of the grafts was small at best. SCs were found at 1 and 2 weeks posttransplant. However, few SCs were found at 2 months posttransplant. Further investigation is under way to characterize the immune capabilities of SCs in a xenogeneic environment. PMID- 16358410 TI - Seeding of corneal wounds by epithelial cell transfer from micropatterned PDMS contact lenses. AB - Persistent corneal wounds result from numerous eye disorders, and to date, available treatments often fail to accelerate reepithelialization, the key initial step in wound healing. To speed reepithelialization, we explored a cell transfer transplant method utilizing polydimethylsiloxane (PDMS) contact lenses to deliver epithelial cells derived from limbal explants directly within a corneal wound. Human primary epithelial cells and an immortalized corneal epithelial cell line (HCE-SV40) grew well on PDMS contact lenses and their morphology and growth rates where similar to cells grown on tissue culture polystyrene. To initially study cell transfer from PDMS, HCE-SV40 cells were seeded onto PDMS with or without micropatterned posts. After a day in culture, HCE-SV40 cells attached to the unpatterned PDMS uniformly, whereas on micropatterned PDMS they appeared to attach primarily between posts. The cell covered PDMS contacts were then placed cell-side down onto tissue culture plastic and, after 1, 2, or 3 days, the PDMS contact was removed and the transferred cells were trypsinized and counted. Micropatterned PDMS contact lenses with 100 microm-diameter posts and a post height of 40 microm transferred three times as many cells as unpatterned PDMS. Cell transfer to a wounded cornea was tested in a pig cornea organ culture model de-epithelialized by alkali treatment. Post micropatterned PDMS contact lenses were seeded with labeled HCE-SV40 cells at a density 50,000 cells/cm2 and applied to the wounded pig corneas. After 24, 48, or 96 h of application, PDMS contact lenses were removed, corneas fixed with formaldehyde, and sectioned. After 48 h, epithelial cells transferred from post micropatterned contact lenses to provide 35% epithelial coverage of denuded pig corneas; after 96 h coverage was 65%. We conclude that cell transfer from epithelial-coated PDMS contact lenses micropatterned with posts provides a promising approach to reepithelialize corneal surfaces. PMID- 16358411 TI - [Intervention of liuwei dihuang pill on lupus nephropathy treated with cylophosphamide and glucocorticoids]. AB - OBJECTIVE: To investigate the intervening effect of Liuwei Dihuang pill (LWDH) on lupus nephropathy (LN) treated with glucocorticoids and cyclophosphamide (CTX). METHODS: Sixty-four patients were randomly divided into two groups, all patients were treated by conventional treatment, using prednisone in standard program, and CTX in a daily dose of 8 - 12 mg/kg, accumulated dose < or = 150 mg/kg, by adding into 500 ml of 5% glucose in saline through intravenous dripping, as well as the symptomatic treatment. Patients in the treated group were given LWDH additionally. RESULTS: The curative effect in the treated group was significantly superior to that in the control group (P < 0.05). Laboratory indexes, including urinary protein, plasma protein and serum creatinine (SCr), erythrocyte sedimentation rate (ESR), complement C3, etc. were significantly improved in both groups (P < 0.01), but all the improvement, except that of SCr, in the treated group were superior to those in the control group respectively (P <0.05 or P <0.01). Besides, the recurrent rate and incidence rate of adverse reaction in the treated group was significant lower than those in the control group (P < 0.05 and P < 0.01). CONCLUSION: LWDH can significantly enhance the therapeutic effect of CTX and glucocorticoids on LN, decrease the recurrence and shows advantage in counteracting against the adverse effects of glucocorticoids and CTX. PMID- 16358412 TI - Where are linear feature extraction methods applicable? AB - A fundamental problem in computer vision and pattern recognition is to determine where and, most importantly, why a given technique is applicable. This is not only necessary because it helps us decide which techniques to apply at each given time. Knowing why current algorithms cannot be applied facilitates the design of new algorithms robust to such problems. In this paper, we report on a theoretical study that demonstrates where and why generalized eigen-based linear equations do not work. In particular, we show that when the smallest angle between the ith eigenvector given by the metric to be maximized and the first i eigenvectors given by the metric to be minimized is close to zero, our results are not guaranteed to be correct. Several properties of such models are also presented. For illustration, we concentrate on the classical applications of classification and feature extraction. We also show how we can use our findings to design more robust algorithms. We conclude with a discussion on the broader impacts of our results. PMID- 16358413 TI - Effects of blood donation on arterial blood pressure in retired racing Greyhounds. AB - The purpose of this study was to evaluate changes in systolic arterial blood pressure (SABP) immediately after collection of blood for transfusion in retired racing Greyhounds. We prospectively evaluated 19 blood donor Greyhounds before and after the collection of a unit (450 mL) of blood. The SABP was measured with Doppler in the right forearm after the dogs had been in the blood collection room for a few minutes (PRE-FLOOR) and again 5-10 minutes after the dogs were placed on the table where they would be bled (PRE-TABLE). A total of 3-5 minutes after completing the blood collection, the SABP was measured again while the dogs were still in lateral recumbency on the table (POST-TABLE) and once more 60-90 minutes later, when the dogs were on the floor after completing the donation (POST FLOOR). All dogs were monitored for clinical signs of hypotension, including depression, weakness, collapse, and pallor, for a minimum of 2 hours after donation. There was a significant difference in SABP for the group between PRE FLOOR and POST-TABLE (P = .02) and between PRE-TABLE and POST-TABLE determinations (P = .01). There were no significant differences for any of the other time points; there were no adverse events. Therefore, we conclude that the collection of 450 mL of blood from normal Greyhounds results in a short-lived yet significant decrease in SABP, but the likelihood of adverse events is negligible. PMID- 16358414 TI - Role of arginine residues 14 and 15 in dictating DNA binding stability and transactivation of the aryl hydrocarbon receptor/aryl hydrocarbon receptor nuclear translocator heterodimer. AB - The aryl hydrocarbon receptor (AHR) and its DNA binding partner, the aryl hydrocarbon receptor nuclear translocator (ARNT) are basic helix-loop-helix/PAS proteins. The goal of the current study was to determine the extent to which residues R14 and R15 contained within the basic region of the AHR contribute to the DNA binding affinity and stability of the AHR/ARNT heterodimer. Towards this end, we first performed equilibrium binding and dissociation rate analyses using a single dioxin response element (DRE-1). While the K(D) and Bmax values obtained from the equilibrium binding analysis were similar for the wild-type AHR (wt AHR) and that containing the substitutions of R14 and R15 with Q residues (Q14Q15 AHR), dissociation rate analyses revealed that the stability of the Q14Q15 AHR DNA binding complex was approximately 10-fold less. Using a two-site DNA binding model, we also found that AHR/ARNT heterodimer does not participate in cooperative binding, as binding of the second dimer appears to be prohibited by occupation of the first. This property was similar regardless of the composition of the amino acids at positions 14 and 15. Finally, reporter assays revealed that the Q14Q15 substitutions severely compromised the ability of the AHR to activate gene expression despite appropriate nuclear localization. The present results revealed that DNA binding stability of the AHR/ARNT heterodimer is an important requirement for its transactivation capabilities and that this stability is governed, in part, by residues R14 and R15 that lie within the basic region of the AHR. PMID- 16358415 TI - The association of TIF-IA and polymerase I mediates promoter recruitment and regulation of ribosomal RNA transcription in Acanthamoeba castellanii. AB - Large amounts of energy are expended for the construction of the ribosome during both transcription and processing, so it is of utmost importance for the cell to efficiently regulate ribosome production. Understanding how this regulation occurs will provide important insights into cellular growth control and into the coordination of gene expression mediated by all three transcription systems. Ribosomal RNA (rRNA) transcription rates closely parallel the need for protein synthesis; as a cell approaches stationary phase or encounters conditions that negatively affect either growth rate or protein synthesis, rRNA transcription is decreased. In eukaryotes, the interaction of RNA polymerase I (pol I) with the essential transcription initiation factor IA (TIF-IA) has been implicated in this downregulation of transcription. In agreement with the first observation that rRNA transcription is regulated by altering recruitment of pol I to the promoter in Acanthamoeba castellanii, we show here that pol I and an 80-kDa homologue of TIF-IA are found tightly associated in pol I fractions competent for specific transcription. Disruption of the pol I-TIF-IA complex is mediated by a specific dephosphorylation of either pol I or TIF-IA. Phosphatase treatment of TIF-IA containing A. castellanii pol I fractions results in a downregulation of both transcriptional activity and promoter binding, reminiscent of the inactive pol I fractions purified from encysted cells. The fraction of pol I competent for promoter recruitment is enriched in TIF-IA relative to that not bound by immobilized promoter DNA. This downregulation coincides with an altered electrophoretic mobility of TIF-IA, suggesting at least it is phosphorylated. PMID- 16358416 TI - Wound site neutrophil transcriptome in response to psychological stress in young men. AB - Communication between the central nervous and the immune system occurs through chemical messengers secreted by nerve cells, endocrine organs, or immune cells. Psychological stressors can disrupt these networks. We have previously observed that disruption of the neuroendocrine immune system adversely influences a broad range of physiological processes including wound healing. Migration of neutrophils to the wound site is an early event that induces a transcriptional activation program, which regulates cellular fate and function, and promotes wound healing. In this study, we have sought to identify stress-sensitive transcripts in wound site neutrophils. A skin blister model was used to collect wound fluid and wound site neutrophils from four young men, experiencing or not examination stress. Self-reported stress was recorded using the Beck Depression Inventory. Stress decreased growth hormone levels at the wound site and was related to impaired wound healing in all subjects. High density microarray analyses were performed using RNA from wound site neutrophils. Results show that psychological stress had an overall suppressive effect on the neutrophil transcriptome. Of the 22,283 transcripts screened, 0.5% were downregulated whereas only under 0.3% were induced by stress in all four out of four subjects. Functionally, stress tilted the genomic balance towards genes encoding proteins responsible for cell cycle arrest, death, and inflammation. Further effort to gain a more comprehensive understanding of the functional significance of such behavior-genome interaction is warranted. PMID- 16358417 TI - A selective role for MRF4 in innervated adult skeletal muscle: Na(V) 1.4 Na+ channel expression is reduced in MRF4-null mice. AB - The factors that regulate transcription and spatial expression of the adult skeletal muscle Na+ channel, Na(V) 1.4, are poorly understood. Here we tested the role of the transcription factor MRF4, one of four basic helix-loop-helix (bHLH) factors expressed in skeletal muscle, in regulation of the Na(V) 1.4 Na+ channel. Overexpression of MRF4 in C2C12 muscle cells dramatically elevated Na(V) 1.4 reporter gene expression, indicating that MRF4 is more efficacious than the other bHLH factors expressed at high levels endogenously in these cells. In vivo, MRF4 protein was found both in extrajunctional and subsynaptic muscle nuclei. To test the importance of MRF4 in Na(V) 1.4 gene regulation in vivo, we examined Na+ channel expression in MRF4-null mice using several techniques, including Western blotting, immunocytochemistry, and electrophysiological recording. By all methods, we found that expression of the Na(V) 1.4 Na+ channel was substantially reduced in MRF4-null mice, both in the surface membrane and at neuromuscular junctions. In contrast, expression of the acetylcholine receptor, and in particular its alpha subunit, was unchanged, indicating that MRF4 regulation of Na+ channel expression was selective. Expression of the bHLH factors myf-5, MyoD, and myogenin was increased in MRF4-null mice, but these factors were not able to fully maintain Na(V) 1.4 Na+ channel expression either in the extrajunctional membrane or at the synapse. Thus, MRF4 appears to play a novel and selective role in adult muscle. PMID- 16358419 TI - Perceiving illumination inconsistencies in scenes. AB - The human visual system is adept at detecting and encoding statistical regularities in its spatiotemporal environment. Here, we report an unexpected failure of this ability in the context of perceiving inconsistencies in illumination distributions across a scene. Prior work with arrays of objects all having uniform reflectance has shown that one inconsistently illuminated target can 'pop out' among a field of consistently illuminated objects (eg Enns and Rensink, 1990 Science 247 721 723; Sun and Perona, 1997 Perception 26 519-529). In these studies, the luminance pattern of the odd target could be interpreted as arising from either an inconsistent illumination or inconsistent pigmentation of the target. Either cue might explain the rapid detection. In contrast, we find that once the geometrical regularity of the previous displays is removed, the visual system is remarkably insensitive to illumination inconsistencies, both in experimental stimuli and in altered images of real scenes. Whether the target is interpreted as oddly illuminated or oddly pigmented, it is very difficult to find if the only cue is deviation from the regularity of illumination or reflectance. Our results allow us to draw inferences about how the visual system encodes illumination distributions across scenes. Specifically, they suggest that the visual system does not verify the global consistency of locally derived estimates of illumination direction. PMID- 16358418 TI - Lack of the DNA repair enzyme OGG1 sensitizes dopamine neurons to manganese toxicity during development. AB - Onset of Parkinson's disease (PD) and Parkinson-like syndromes has been associated with exposure to diverse environmental stimuli. Epidemiological studies have demonstrated that exposure to elevated levels of manganese produces neuropathological changes localized to the basal ganglia, including neuronal loss and depletions in striatal dopamine content. However, understanding the mechanisms associated with manganese neurotoxicity has been hampered by the lack of a good rodent model. Elevated levels of 8-hydroxy-2'-deoxyguanosine (oxo8dG) have been found in brain areas affected in PD. Whether increased DNA damage is responsible for neuronal degeneration or is a mere epiphenomena of neuronal loss remains to be elucidated. Thus, by using mice deficient in the ability to remove oxo8dG we aimed to determine if dysregulation of DNA repair coupled to manganese exposure would be detrimental to dopaminergic neurons. Wild-type and OGG1 knockout mice were exposed to manganese from conception to postnatal day 30; in both groups, exposure to manganese led to alterations in the neurochemistry of the nigrostriatal system. After exposure, dopamine levels were elevated in the caudate of wild-type mice. Dopamine was reduced in the caudate of OGG1 knockout mice, a loss that was paralleled by an increase in the dopamine index of turnover. In addition, the reduction of dopamine in caudate putamen correlated with the accumulation of oxo8dG in midbrain. We conclude that OGG1 function is essential in maintaining neuronal stability during development and identify DNA damage as a common pathway in neuronal loss after a toxicological challenge. PMID- 16358420 TI - The role of edges in object recognition by pigeons. AB - In three experiments, we explored how pigeons use edges, corresponding to orientation and depth discontinuities, in visual recognition tasks. In experiment 1, we compared the pigeon's ability to recognize line drawings of four different geons when trained with shaded images. The birds were trained with either a single view or five different views of each object. Because the five training views had markedly different appearances and locations of shaded surfaces, reflectance edges, etc, the pigeons might have been expected to rely more on the orientation and depth discontinuities that were preserved over rotation and in the line drawings. In neither condition, however, was there any transfer from the rendered images to the outline drawings. In experiment 2, some pigeons were trained with line drawings and shaded images of the same objects associated with the same response (consistent condition), whereas other pigeons were trained with a line drawing and a shaded image of two different objects associated with the same response (inconsistent condition). If the pigeons perceived any correspondence between the stimulus types, then birds in the consistent condition should have learned the discrimination more quickly than birds in the inconsistent condition. But, there was no difference in performance between birds in the consistent and inconsistent conditions. In experiment 3, we explored pigeons' processing of edges by comparing their discrimination of shaded images or line drawings of four objects. Once trained, the pigeons were tested with planar rotations of those objects. The pigeons exhibited different patterns of generalization depending on whether they were trained with line drawings or shaded images. The results of these three experiments suggest that pigeons may place greater importance on surface features indicating materials, such as food or water. Such substances do not have definite boundaries cued by edges which are thought to be central to human recognition. PMID- 16358421 TI - [Temporal variation of soil respiration on sloping pasture of Heihe River basin and effects of temperature and soil moisture on it]. AB - Employing LiCor 6400 gas exchange analyzer and soil respiration chamber attachment (LiCor Inc., Lincoln, NE, USA), this paper continuously measured the soil surface CO2 effluxes on the sloping pasture of Heihe River basin from early April to late October 2003 to investigate the soil CO2 efflux rate and its feedback to the changes of climate and land use. The results showed that from May to October, the diurnal variation of soil respiration was low at night, the lowest at 7:00, 6:30, 5:30, 6:00 and 7:00, raised rapidly at 7:00 - 8:30, and then descended at 16:00 - 18:30. The maximum soil CO2 efflux appeared at 15:00, 14:30, 14:30, 13:30, 14:00 and 15:00. The mean daily soil respiration rate was 0.31 - 6.98 micromol m(-2) s(-1), with the maximum in July and August, the second in May and September, and nearly consistent in April and October. Soil respiration rate had an exponential and power correlation with temperature and soil moisture, respectively. PMID- 16358422 TI - Host range, amplification and arboviral disease emergence. AB - Etiologic agents of arboviral diseases are primarily zoonotic pathogens that are maintained in nature in cycles involving arthropod transmission among a variety of susceptible reservoir hosts. In the simplest form of human exposure, spillover occurs from the enzootic cycle when humans enter zoonotic foci and/or enzootic amplification increases circulation near humans. Examples include Eastern (EEEV) and Western equine encephalitis viruses (WEEV), as well as West Nile (WNV), St. Louis encephalitis (SLEV) and Yellow fever viruses. Spillover can involve direct transmission to humans by primary enzootic vectors (e.g. WNV, SLEV and WEEV) and/or bridge vectors with more catholic feeding preferences that include humans (e.g. EEEV). Some viruses, such as Rift Valley fever, Japanese encephalitis and Venezuelan equine encephalitis viruses (VEEV) undergo secondary amplification involving replication in livestock animals, resulting in greater levels of spillover to humans in rural settings. In the case of VEEV, secondary amplification involves equines and requires adaptive mutations in enzootic strains that allow for efficient viremia production. Two of the most important human arboviral pathogens, Yellow fever and dengue viruses (DENV), have gone one step further and adopted humans as their amplification hosts, allowing for urban disease. The ancestral forms of DENV, sylvatic viruses transmitted among nonhuman primate reservoir hosts by arboreal mosquitoes, adapted to efficiently infect the urban mosquito vectors Aedes aegypti and Ae. albopictus during the past few thousand years as civilizations arose. Comparative studies of the sylvatic and urban forms of DENV may elucidate the evolution of arboviral virulence and the prospects for DENV eradication should effective vaccines be implemented. PMID- 16358423 TI - Control of arbovirus diseases: is the vector the weak link? AB - Arthropod-borne virus (arbovirus) diseases (ABVDs) remain major threats to human health and well-being and, as an epidemiologic group, inflict an unacceptable health and economic burden on humans and animals, including livestock. The developed world has been fortunate to have escaped much of the burden that arboviruses and their arthropod vectors inflict on humans in disease endemic countries, but the introduction and rapid spread of West Nile virus in the Western Hemisphere demonstrated that we can no longer be complacent in the face of these emerging and resurging vector-borne diseases. Unfortunately, as the burdens and threats of ABVDs have increased, the U.S. and international public health capacity to address them has decreased. Vaccines are not available for most of these agents. Previously successful strategies to control ABVDs emphasized vector control, but source reduction and vector control strategies using pesticides have not been sustainable. New insights into vector biology and vector pathogen interactions, and the novel targets that likely will be forthcoming in the vector post-genomics era, provide new targets and opportunities for vector control and disease reduction programs. These findings and approaches must be incorporated into existing strategies if we are to control these important pathogens. PMID- 16358424 TI - The spread of the H5N1 bird flu epidemic in Asia in 2004. AB - H5N1 avian influenza has spread to eight countries in eastern Asia including China, Japan, South Korea, Vietnam, Laos, Cambodia, Thailand, and Indonesia in early 2004. This H5N1 influenza A virus is extremely virulent in poultry including chickens and ducks, killing millions of birds throughout the region. Additionally this virus has transmitted to humans (mainly children) in Vietnam, Cambodia, and Thailand, killing 54 of 100 diagnosed persons. To control this epidemic hundreds of millions of chickens and ducks have been culled. One genotype of H5N1 designated "Z" has become dominant in Asia. This virus was first detected in wild birds in Hong Kong in November 2002 and was antigenically distinct from H5N1 viruses isolated from 1997 to early 2002 and lethal for aquatic birds. The H5N1 virus infecting humans and poultry in Asia in 2004 is an antigenic variant of the Z genotype. Here we consider the possible role of migrating birds in the evolution and spread of the H5N1 influenza A virus throughout Asia. We conclude that the available information is consistent with a role for migrating birds but limited information is available and that serological studies are urgently needed on migrating birds worldwide. The prospect is that this H5N1/04 influenza A virus will become endemic in poultry in eastern Asia and will be a continuing threat to animal and human health. It is also projected that a human H5N1 vaccine will eventually be needed. PMID- 16358425 TI - Ehrlichia under our noses and no one notices. AB - Ehrlichia chaffeensis, an obligately intracellular bacterium, resides within a cytoplasmic vacuole in macrophages, establishes persistent infection in natural hosts such as white-tailed deer and canids, and is transmitted transstadially and during feeding by ticks, particularly Amblyomma americanum. Ehrlichial cell walls contain glycoproteins and a family of divergent 28 kDa proteins, but no peptidoglycan or lipopolysaccharide. The dense-cored ultrastructural form preferentially expresses certain glycoproteins, including a multiple repeat unit containing adhesin. Ehrlichiae attach to L-selectin and E-selectin, inhibit phagolysosomal fusion, apoptosis, and JAK/STAT activation, and downregulate IL 12, IL-15, IL-18, TLR2 and 3, and CD14. Mouse models implicate overproduction of TNF-alpha by antigen-specific CD8 T lymphocytes in pathogenesis and strong type 1 CD4 and CD8 T lymphocyte responses, synergistic activities of IFN-gamma and TNF alpha, and IgG2a antibodies in immunity. Human monocytotropic ehrlichiosis (HME) manifests as a flu-like illness that progresses in severity to resemble toxic shock-like syndrome, with meningoencephalitis or adult respiratory distress syndrome in some patients, and requires hospitalization in half. In immunocompromised patients, HME acts as an overwhelming opportunistic infection. In one family physician's practice, active surveillance for three years revealed an incidence of 1000 cases per million population. Diagnosis employs serology or polymerase chain reaction, which are not utilized sufficiently to establish the true impact of this emerging virus-like illness. PMID- 16358426 TI - Structural biology of old world and new world alphaviruses. PMID- 16358427 TI - Trabecular meshwork alteration and intraocular pressure change following pulsed near-infrared laser trabeculoplasty in cats. AB - BACKGROUND AND OBJECTIVE: To comparatively assess the safety and variation in intraocular pressure (IOP) of two pulsed near-infrared lasers (titanium:sapphire and alexandrite) for laser trabeculoplasty versus conventional blue-green argon laser trabeculoplasty in an animal model. MATERIALS AND METHODS: The left eyes of 15 healthy cats received a 180 degree laser trabeculoplasty treatment: 5 with a titanium:sapphire laser, 5 with an alexandrite laser, and 5 with an argon laser. Preoperatively and postoperatively, all animals underwent tonometry, gonioscopy, and slit-lamp examination. The cats were observed up to 12 weeks. Scanning electron microscopy and histologic examination were performed to evaluate potential alterations in the trabecular meshwork structure. RESULTS: IOP at 1 hour, 1 day, and 1 week following treatment was remarkably lower, irrespective of the laser source used. Following treatment with both near-infrared lasers, gonioscopy showed depigmentation underneath the area of the treated trabecular meshwork and histologic evaluation showed a decrease in pigment density. On scanning electron microscopy, damage to the trabecular meshwork structure could not be detected after treatment with near-infrared lasers. CONCLUSIONS: Near infrared laser trabeculoplasty was found to be effective to temporarily lower IOP in cats. The lasers selectively altered pigment-containing cells, avoiding structural damage of the trabecular meshwork anatomy. PMID- 16358428 TI - Short-term metabolic ethanol tolerance in dogs. AB - Metabolic ethanol tolerance was studied in a cohort of five dogs with ethanol challenge repeated weekly over a 7-week period. During the 7-week period, the area under the blood alcohol versus time curve (AUC) increased slightly while the rate of ethanol elimination also increased slightly. During the repeated ethanol dosing, ethanol absorption shifted from approximately equal absorption in the stomach and intestine to three-fold more absorption in the intestine than in the stomach. The likely cause of the shift in absorption site was probably a concomitant change in gastric emptying that occurred with repeated dosing. This shift is significant since ethanol absorption in the small intestine has been shown to be over six-fold more rapid than ethanol absorption in the stomach. PMID- 16358429 TI - [Convulsions in association with bupropion]. PMID- 16358430 TI - [Thrombotic thrombocytopenic purpura as presentation of a neoplasia]. PMID- 16358431 TI - [Telangiectasia macularis eruptiva Perstans and Conn syndrome]. PMID- 16358432 TI - [Brucella epididymo-orchitis]. PMID- 16358433 TI - [Inguinal mass as presentation of cat-scratch disease]. PMID- 16358434 TI - [Community acquired pneumonia and explosive-pleuritis]. PMID- 16358435 TI - [Pericardial effusion infected by Salmonella enteritidis]. PMID- 16358436 TI - [Interstitial pneumonitis as a clinical manifestation in a patient with systemic sclerosis without scleroderma]. PMID- 16358437 TI - [Eosinophilia in patients infected with the human immunodeficiency virus]. PMID- 16358438 TI - The inheritance of the core self in human clones. PMID- 16358439 TI - Which IVF test is best for baby? PMID- 16358440 TI - Not by the book. PMID- 16358441 TI - Introduction: ethical responsibilities regarding drugs, patents, and health. PMID- 16358442 TI - Intellectual property and pharmaceutical drugs: an ethical analysis. AB - The pharmaceutical industry has in recent years come under attack from an ethical point of view concerning its patents and the non-accessibility of life-saving drugs for many of the poor in both less developed countries and in the United States. The industry has replied with economic and legal justifications for its actions. The result has been a communication gap between the industry on the one hand and poor nations and American critics on the other. This paper attempts to present and evaluate the arguments on all sides and suggests a possible way out of the current impasse. It attempts to determine the ethical responsibility of the drug industry in making drugs available to the needy, while at the same time developing the parallel responsibilities of individuals, governments, and NGOs. It concludes with the suggestion that the industry develop an international code for its self-regulation. PMID- 16358443 TI - The corporate social responsibility of the pharmaceutical industry: idealism without illusion and realism without resignation. AB - In recent years society has come to expect more from the "socially-responsible" company and the global HIV/AIDS pandemic in particular has resulted in some critics saying that the "Big Pharma" companies have not been living up to their social responsibilities. Corporate social responsibility can be understood as the socio-economic product of the organizational division of labor in complex modern society. Global poverty and poor health conditions are in the main the responsibilities of the world's national governments and international governmental organizations, which possess society's mandate and appropriate organizational capabilities. Private enterprises have neither the societal mandate nor the organizational capabilities to feed the poor or provide health care to the sick in their home countries or in the developing world. Nevertheless, private enterprises do have responsibilities to society that can be categorized as what they must do, what they ought do, and what they can do. PMID- 16358444 TI - Intellectual property rights, moral imagination, and access to life-enhancing drugs. AB - Although the idea of intellectual property (IP) rights--proprietary rights to what one invents, writes, paints, composes or creates--is firmly embedded in Western thinking, these rights are now being challenged across the globe in a number of areas. This paper will focus on one of those challenges: government sanctioned copying of patented drugs without permission or license of the patent owner in the name of national security, in health emergencies, or life threatening epidemics. After discussing standard rights-based and utilitarian arguments defending intellectual property we will present another model. IP is almost always a result of a long history of of scientific or technological development and numbers of networks of creativity, not the act of a single person or a group of people at one moment in time. Thus thinking about and evaluating IP requires a traditional model of IP. It follows that the owner of those rights has some obligations to share that information or its outcomes. If that conclusion is applied to the distribution of antiretroviral drugs, what pharmaceutical companies are ethically required to do to increase access to these medicines in the developing world will have to be reanalyzed from a more systemic perspective. PMID- 16358445 TI - Unusually high alkaline phosphatase due to intestinal isoenzyme in a healthy adult. AB - Intestinal alkaline phosphatase (ALP) is more prevalent in individuals of blood group O or B, and increases after a meal, especially on a high-fat diet. We did not realize that clinicians could underestimate the importance of fasting for ALP measurement until one healthy adult showed a huge difference in ALP levels before and after a meal. We report a case of transient hyperphosphatasemia resulting in unnecessary workup due to intestinal ALP in a healthy adult of blood group O. PMID- 16358446 TI - Swedish guidelines for the management of community-acquired pneumonia in immunocompetent adults. AB - This document presents the evidence-based guidelines of the Swedish Society of Infectious Diseases for the management of adult immunocompetent patients with community-acquired pneumonia (CAP), who are assessed at hospital. The prognostic score 'CURB-65' is recommended for all CAP patients in the emergency room. The score provides an assessment tool for the decision regarding outpatient treatment or level of hospital supervision, the choice of microbiological investigations, and empirical antibiotic treatment. In patients with non-severe CAP (CURB-65 score 0-2) we recommend initial narrow-spectrum antibiotic treatment, orally or intravenously, primarily directed at Streptococcus pneumoniae. In those with CURB 65 score 3, penicillin G or a cephalosporin intravenously is recommended. For CURB-65 score 0-3 atypical pathogens should be covered only when they are suspected on clinical or epidemiological grounds. In patients with CURB-65 score 4-5 intravenous combination therapy with either cephalosporin/macrolide or penicillin G/fluoroquinolone is recommended. Efforts should be made to identify the CAP aetiology in order to support the ongoing antibiotic treatment or to suggest treatment alterations. Recommended measures for prevention of CAP include influenza -- and pneumococcal -- vaccination to risk groups and efforts for smoking cessation. PMID- 16358447 TI - The management of infections in children in general practice in Sweden: a repeated 1-week diagnosis-prescribing study in 5 counties in 2000 and 2002. AB - A diagnosis-prescribing study was performed in 5 Swedish counties during 1 week in November in 2000 and repeated in 2002. The aim of the present study was to analyse data for children 0-15y of age who consulted a general practitioner with symptoms of an infection. During the 2 weeks studied, 4049 children were consulted. Respiratory tract infections (RTI) were the predominant diagnoses, above all among the youngest children, while the proportion of urinary tract infections and skin infections increased with increasing age. Between the y 2000 and 2002, the proportion of children allocated the diagnosis streptococcal tonsillitis and pneumonia decreased (p<0.01 and p<0.001, respectively) while the proportion of common cold increased (p<0.001). Antibiotic prescribing decreased from 55% to 48% (p<0.001) for respiratory infections between the years studied. The only significant changes in type of antibiotics prescribed were the increase of isoxazolylpenicillins (p<0.001) used for skin infection and the decrease of macrolides (p=0.001). A diagnostic test was used in more than half of the consultations. Of children allocated a RTI diagnosis, 36% were prescribed antibiotics when a C-reactive protein test was performed compared to 58% in those not tested. Further studies are needed in general practice to determine the optimal use of near-patient tests in children with RTI. PMID- 16358448 TI - Brief report: respiratory syncytial virus activity--United States, 2004-2005. AB - Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections (LRTIs) (e.g., bronchiolitis and pneumonia) among young children, resulting in an estimated 51,000-82,000 hospitalizations annually in the United States. RSV also causes severe disease and death among older persons and persons of all ages with compromised respiratory, cardiac, or immune systems and can exacerbate chronic cardiac and pulmonary conditions. In temperate climates, most RSV infections occur during a distinct seasonal peak. This report presents preliminary data from RSV activity reported to the National Respiratory and Enteric Virus Surveillance System (NREVSS) for the weeks ending July 2 through December 3, 2005, indicating the onset of the 2005-06 RSV season, and summarizes trends during July 2004-June 2005. Health-care providers should consider RSV in the differential diagnosis for persons of all ages with LRTIs, implement appropriate isolation precautions to prevent nosocomial transmission, and provide appropriate immune prophylaxis to eligible children, including certain premature infants or infants and children with chronic lung and heart disease. PMID- 16358449 TI - Thai Chi may be safe though ineffective for rheumatoid arthritis. PMID- 16358450 TI - Spinal fusion slightly more effective than intensive rehabilitation for chronic low back pain. PMID- 16358451 TI - Intensive rehabilitation may be more cost effective than surgical stabilization for chronic low back pain. PMID- 16358452 TI - Oswestry Disability Questionnaire (ODQ). PMID- 16358453 TI - The Neck Disability Index (NDI). PMID- 16358454 TI - [Administration of aprotinin in surgical interventions associated with a high risk hemorrhagic complications]. PMID- 16358455 TI - [Perforation of a fibrous capsule of echinococcal cyst]. PMID- 16358456 TI - [Perforation of the stomach in generalized tuberculosis]. PMID- 16358457 TI - [Leiomyosarcoma of the ascendant part of the duodenum with decompensated duodenostasis]. PMID- 16358458 TI - [Natural dyes and dyeing from the XVIIth century to the birth of synthetic dyes]. AB - After historical considerations on the state and evolution of French dyeing industry in the end of the XVIIth century to the beginning of XIXth, we find this presentation a resume of the different states of tissues dyeing. We easily note that the techniques of dyeing. We easily note that the techniques of dyeing were very brought forward the end of the XVIIIth century before that synthetic dyes appeared in the second half of the XIXth century. PMID- 16358459 TI - [Is Chevreul accountable for the backward state of synthetic dyes in France?]. AB - Head of dying at the Manufacture des Gobelins, Chevreul presided over the French Council of Arts and Manufactures and took part in the selection committees of universal exhibitions. He was accused of lacking judgement against synthetic dyes. His correspondence with dyers shows other more determinant brakes as legislative and economic ones, against which scientist, without political power, could not act efficiently. PMID- 16358460 TI - [Injectable dyes: more than 70 years of therapeutic usage of dyes]. AB - Synthetic dyes were first discovered at the end of the XVIIIth century (picric acid by Woulfe in 1771) and during the first part of the XIXth century: rosolic acid (Runge, 1834), mauvein (Perkin, 1856), but their development was major at the end of the XIXth century. Their therapeutic use was really starting at the beginning of the XXth century, mainly by local and oral route. It is only in the 1930's that parenteral route of administration was actually developed: methylene blue, methyl violet are some examples that were used for leprosy and filariasis, respectively. After the 2nd world war, one can observe a rapid decrease of the therapeutic use of dyes by intravenous route. Only a few dyes are still used today, such as patent blue V or fluorescein, as drugs for diagnostic use. PMID- 16358461 TI - Sun, sand, and an M.D. PMID- 16358462 TI - MIT fires biology professor who admitted faking data. PMID- 16358463 TI - Report knocks agriculture department's protection of research animals. PMID- 16358464 TI - [The evolution of the vocational training of apothecaries and the pharmacists through the << Salle des Actes>> of the faculty of pharmacy of Paris]. AB - The visit of la <> of the faculty of pharmacy of Paris brings important knowledge on the evolution of the vocational training of the apothecaries and the pharmacists during nearly five centuries. The characters represented on the portraits played a major role in the formation and dissemination of knowledge. PMID- 16358465 TI - Call to revolution in the prevention of emotional disorders. AB - This article argues that it is futile to try and treat the many serious mental disturbances now confronting society with individual psychotherapy. Only by preventing the occurrence of mental disturbance can headway be made in dealing with this source of human suffering. Prevention means confronting and ameliorating the major cause of mental disturbance: stress. The sources of stress include poverty, rampant sexism, being born unwanted, and other forms of social injustice. This article demands that those in the mental health field examine their moral and scientific beliefs and begin to take steps to change the existing unfair political and economic practices. PMID- 16358466 TI - [The mineralogical and petrographical work of the pharmacists in the Museum]. AB - Three famous pharmacists were working in the Museum national d'histoire naturelle in Paris, in the field of earth sciences: Louis Nicolas Vauquelin (1763-1829), Andre Laugier (1770-1832) and Alfred Lacroix (1863-1948). Vauquelin, professor of Chemical arts, established the chemical composition of numerous minerals, which led him to the discovery of new chemical elements. He also took a hand in demonstrating the extra-terrestrial origin of meteorites. Laugier, professor of General chemistry, was an admitted expert in the analytical field of these rocks. Lacroix, who held during more than forty years the chair of Mineralogy in the Museum, carried out major scientific work. This field working naturalist who was also famous vulcanologist placed his studies of the mineral species into a petrographical context. He described numerous new minerals, took an interest in their origin and classification of the volcanic rocks, as well as in contact metamorphism. In other respects, he increased the collections in the Museum (minerals, rocks, meteorites). The works of Vauquelin, Laugier and Lacroix contributed to advance those begun by some scientists of the Jardin du Roi, as Buffon or the Rouelle brothers before the French Revolution. PMID- 16358467 TI - Psychotherapy as civics: the patient therapist as citizens. AB - The field of psychotherapy has failed to live up to its promise to be a force in the continued development of democracy by aligning itself with medicine and psychiatry. Psychotherapists must recognize that those with whom they work and disseminate information to are inevitably citizens and that they are redefining citizens as mental patients. By utilizing psychiatric diagnoses, which are in effect, moral labels, psychotherapy unwittingly helps create authoritarian political structures by convincing their patients and the public that they are essentially inferior and can do nothing to change that condition. A series of recommendations are made that suggest ways for psychotherapy to free itself from institutional psychiatry and medicine and build its own "house." PMID- 16358468 TI - Changes in LHbeta-gene and FSHbeta-gene expression in the ram pars tuberalis according to season and castration. AB - Luteinizing hormone beta (LHbeta) and follicle stimulating hormone beta (FSHbeta) subunits and their mRNAs were studied in the ram pars tuberalis following different seasonal (winter vs summer) and experimental (intact vs castrated animals) conditions. Hormone-containing cells were identified by immunohistochemistry, using homologous double-stranded 35S-cDNAs. The labelling was quantified by image analysis. Immunohistochemical staining showed that cells containing LHbeta and FSHbeta were localized mainly in the ventral part of the pars tuberalis but that, in the summer, additional LHbeta containing cells were present in the dorsal part in intact rams. On the other hand, LHbeta-mRNA labelling was found in the whole pars tuberalis in wethers but only in the ventral part in intact rams. The magnitude of LHbeta-mRNA labelling was significantly greater in summer than in winter rams, and in castrated than in intact animals (P<0.001). However, the number of labelled cells was found to be the greatest in the winter (P<0.001) and was not affected by castration. FSHbeta mRNA expression was similar to that of LHbeta-mRNA except that the level and extent were considerably lower. Thus, our results show an increase in the magnitude of gonadotropin beta subunit-mRNA in the summer and following castration; this increase appears to involve the entire pars tuberalis. PMID- 16358469 TI - Reification of psychiatric diagnoses as defamatory: implications for ethical clinical practice. AB - While the mental health professional generally has beneficent motives and an honest belief in the DSM diagnoses assigned to clients, such diagnoses may yet be defamatory when communicated to third parties. Mental health diagnoses invariably lower the individual's reputation in the eyes of the community. At the same time, DSM diagnoses are but one out of a myriad of possible interpretive frameworks. DSM descriptors for the client's distress thus cannot be said to capture the essence of the client's personhood. When a diagnosis is published as if it captured a definitive truth about an individual psychiatric client, it is, in that important regard, inaccurate. That is, such a communication meets the criterion for a reckless disregard for the truth or an honest belief but without reasonable basis insofar as it is considered to be anything more than a working hypothesis. Hence, in certain cases, DSM labeling may constitute defamation. PMID- 16358470 TI - Life: medicine, morals, and markets. Right...or commodity? PMID- 16358471 TI - The legacy of Terri Schiavo: the disabled sound the alarm for the nonreligious. PMID- 16358472 TI - Making our own decisions about death: competency should be paramount. PMID- 16358473 TI - Determining the value of life: discrimination, advance directives, and the right to die with dignity. PMID- 16358474 TI - Why not a kidney market? An obvious solution to the organ shortage. PMID- 16358475 TI - Fetal pain: a red herring in the abortion debate. PMID- 16358477 TI - Agency proposes major expansion of its regulations for human subjects. PMID- 16358476 TI - Complaint alleges "coercion" and "undue influence" in IRB-approved consent form. PMID- 16358478 TI - Complaint alleges many deficiencies in IRB-approved informed consent form. PMID- 16358479 TI - New federal guidance adds duties for institutional review boards (IRBs). PMID- 16358480 TI - IRBs, human subjects, and reporting adverse events. PMID- 16358481 TI - Complaint alleges that informed consent procedures were used after experiment began. PMID- 16358482 TI - Transgenderism. PMID- 16358483 TI - Does anyone actually invoke their hospital futility policy? PMID- 16358484 TI - Clonally expanded T-cell populations in atomic bomb survivors do not show excess levels of chromosome instability. AB - Radiation-induced genomic instability has been studied primarily in cultured cells, while in vivo studies have been limited. One major obstacle for in vivo studies is the lack of reliable biomarkers that are capable of distinguishing genetic alterations induced by delayed radiation effects from those that are induced immediately after a radiation exposure. Here we describe a method to estimate cytogenetic instability in vivo using chromosomally marked clonal T-cell populations in atomic bomb survivors. The basic idea is that clonal translocations are derived from single progenitor cells that acquired an aberration, most likely after a radiation exposure, and then multiplied extensively in vivo, resulting in a large number of progeny cells that eventually comprise several percent of the total lymphocyte population. Therefore, if chromosome instability began to operate soon after a radiation exposure, an elevated frequency of additional but solitary chromosome aberrations in clonal cell populations would be expected. In the present study, six additional translocations were found among 936 clonal cells examined with the G-band method (0.6%); the corresponding value with multicolor FISH analysis was 1.2% (4/333). Since these frequencies were no higher than 1.2% (219/17,878 cells), the mean translocation frequency observed in control subjects using the G-band method, it is concluded that chromosome instabilities that could give rise to an increased frequency of persisting, exchange-type aberrations were not commonly generated by radiation exposure. PMID- 16358485 TI - [Comparison of different phenotypic methods detecting extended spectrum beta lactamases in Pseudomonas aeruginosa and Acinetobacter baumannii strains]. AB - Pseudomonas aeruginosa and Acinetobacter baumannii which are of great concern as the frequent causative agents of nosocomial infections, frequently exhibit resistance to beta-lactam antibiotics. One of the most common causes of this resistance is the presence of extended spectrum beta-lactamases (ESBL). There is still no standardized method for the detection of ESBLs in P. aeruginosa and A.baumannii strains. The aim of this study was to compare the results of Epsilomer test (E-test), combined disk (CD) and double disk synergy (DDS) methods for the detection of ESBL in a total of 38 P. aeruginosa and 45 A.baumannii strains with ceftazidim or cefotaxime MIC > or = 1 microg/ml, isolated from different clinical samples. Cefepime was included in the group of indicator antibiotics and the distance between clavulanic acid containing disk and indicator antibiotic disks was reduced to increase the sensitivity of DDS test. Of the P. aeruginosa strains, 78.9% was detected as ESBL producers by ceftazidime/ ceftazidime-clavulanic acid and 21.1% detected by cefotaxime/cefotaxime-clavulanic acid ratio. Among this group of strains, 31.5% was positive by DDS test and 84.2% was positive by CD method. Of the A.baumannii strains, 93.3% of them was positive by ceftazidime/ceftazidime-clavulanic acid and 6.6% was positive by cefotaxime/cefotaxime-clavulanic acid ratio. DDS test detected 20% and CD method detected 53.3% of the strains positive by E-test method. The use of cefotaxime together with ceftazidime as an indicator antibiotic increased the sensitivity of detection of ESBL enzymes. Although inclusion of cefepime and reduction of the distance between disks increased the detection of ESBL by DDS method, it was still less effective than E-test. Although, 28.9% of the P. aeruginosa and 13.3% of the A. baumannii strains were susceptible to ceftazidime by in vitro tests, they were ESBL producers. In conclusion, E-test method was found to be the most sensitive phenotypic method for the detection of ESBL in our study collection. PMID- 16358486 TI - [Evaluation of disk diffusion test for carbapenem sensitivity in Acinetobacter baumannii and Pseudomonas aeruginosa strains]. AB - This study was conducted upon the detection of discordant results obtained for Acinetobacter species against meropenem by disk diffusion susceptibility and E test method. The aim of this study was to investigate the reliability of disk diffusion method for the determination of carbapenem susceptibility in non fermentative bacilli. Carbapenem (imipenem and meropenem) sensitivites of 157 Acinetobacter baumanii and 157 Pseudomonas aeruginosa strains were performed by disk diffusion test and E-test in our laboratory where quality control of antibiotic susceptibility tests are performed weekly. For meropenem, discordance of the two tests were 10.8% and 4.5% for A. baumannii and P. aeruginosa, respectively. MIC values and disk diffusion zone diameters were significantly different from each other. There was no difference in terms of imipenem in A. baumannii strains but 5.1% of P. aeruginosa isolates showed discordance. Since, for the P. aeruginosa isolates with discordant imipenem susceptibility results, the imipenem disk diffusion zones were close to the lower level and their MIC values were within the intermediate resistance range, it was assumed that this technical discordance had low risk of reproducibility. In routine laboratories, meropenem resistance detected by disk diffusion method, should be verified by another technique. This is not only important for patient results but for epidemiological data as well. Routine quality control studies for antibiotic susceptibility tests and close observation of suspicious results, is mandatory for each laboratory, in order to achieve good quality performance. PMID- 16358487 TI - [Standardization of an in-house ELISA for pertussis serology and its application in a seroepidemiological study]. AB - Seroepidemiological studies need sensitive, practicle and cost-effective methods. For pertussis serosurveillance, an in-house ELISA for antipertussis-toxin (PT) and anti-filamentous-hemagglutinin (FHA) were established in our laboratory and compared by a Ball-ELISA which had been reported to be reliable previously. Sixty sera with various antibody titers were tested by both of the methods. The correlation coefficients between two methods were 0.729 and 0.776 for anti-PT and anti-FHA, respectively, and regression coefficients were 0.623 and 0.693, respectively. The in-house ELISA was applied to a serosurvey including 373 healthy subjects (6 months-91 years old) in Turkey to observe the results. The moving averages of both antibodies were increased until 10 years old, reaching to 31 EU/ml for anti-PT and 65 EU/ml for anti-FHA and kept around this level in the older ages. The in-house ELISA was found to be reliable and the serosurvey results obtained by ELISA showed a characteristic distribution of antibody titers in each age group. PMID- 16358488 TI - [Comparison of the serological tests used for the laboratory diagnosis of brucellosis]. AB - Brucellosis which is an important public health problem is a zoonotic disease that causes economic loss, and seen all over the world as well as in our country. The aim of this study was to compare the results of Rose Bengal test, standard tube agglutination test (STA), Coombs tube agglutination test, Rivanol tube agglutination test and ELISA (IgA, IgG and IgM) method, in patients who were suspected to have brucellosis. Blood and serum samples collected from 77 patients were included to the study. Blood cultures were performed from all of the patients and Brucella spp were isolated in 35 (45.4%) of them. At least one of the serological tests for Brucella was found positive in 62 samples (80.5%). When culture positivity accepted as the gold standard, the sensitivities of the tests were calculated as follows; 100% for Rose Bengal, 94.3% for STA, 97.1% for ELISA IgG, 94.3 for ELISA IgA and 71.4 for ELISA IgM. No prezone phenomenon was observed in the samples. Brucella Coombs tube agglutination test did not increase the chance of diagnosis of brucellosis, unlike the results of previous studies. We found that Rose Bengal and STA tests were still efficient methods for brucellosis serodiagnosis. The results of rivanol tube agglutination test were similar to STA test, and this test can be used especially in patients with brucellosis relapse. ELISA test detecting IgA and IgG antibody titers together found to be effective methods increasing the chance of brucellosis diagnosis, and could be helpful for follow up of brucellosis. PMID- 16358489 TI - [In vitro activity of ketoconazole, itraconazole and terbinafine against Malassezia strains isolated from neonates]. AB - Malassezia, a yeast-like fungus found in normal skin flora is known to be associated with various skin diseases and systemic infections. There is yet no standardized in vitro susceptibility testing method and minimum inhibitory concentration (MIC) breakpoints for Malassezia species. In this study, we investigated the in vitro activity of ketoconazole, itraconazole and terbinafine against 30 Malassezia strains (22 Malassezia furfur/dermatis, 8 M. japonica) isolated from cheek and/or scalp swabs and/or cephalic pustules of 21 neonates. The isolates were identified to species level on the basis of growth on Sabouraud dextrose agar, assimilation of Tween compounds and catalase reaction. The antifungal susceptibility tests were performed by agar dilution method using modified Dixon agar (MDA) and the agar dilution plates were incubated at 32 degrees C. For the isolates which exhibited sufficient growth, MICs were read at 48 hours, for the remaining slow-growing isolates, MIC readings were done on 7th day. For all drugs tested, the lowest concentration that provided complete inhibition of growth visually was interpreted as the MIC (microg/ml) value. Itraconazole was the most active drug in vitro against Malassezia species, followed by ketoconazole and terbinafine in rank order. In vitro activity of terbinafine was poor for half of the Malassezia strains tested. Variations in activity against individual Malassezia strains were detected for ketoconazole and terbinafine, while in vitro activity of itraconazole was similar for all strains tested. In order to validate the clinical significance of these results, further in vitro and in vivo correlation studies are needed. PMID- 16358490 TI - [Evaluation of species distribution of yeasts isolated from intensive care units during the four years period]. AB - The aim of this study was to evaluate the distributions of yeast species according to the years and to detect the emerging pathogens in intensive care units (ICU). For this purpose, yeast isolation rates were detected retrospectively, in the following time periods: Period I: April-December 2001; period II: January-December 2002; period III: January-December 2003; period IV: January-December 2004. A total of 490 yeast isolates recovered from 462 clinical specimens obtained from 360 different ICU patients were investigated during these periods. Urine (62.1%), blood (13.6%) and tracheal aspirate (8.7%) samples were detected as the most common specimens. Of these isolates, 53.3% were identified as Candida albicans, 14.5% as C. tropicalis, 12.2% as C. glabrata, 6.5% as C. parapsilosis, 4.5% as Trichosporon spp., 3.9% as C. kefyr, 1.6% as C. krusei, 1.4% as Geotrichum candidum and 2.1% as other Candida species. The isolation rates of C. albicans in the periods of I to IV were found as 47.7%, 55.5%, 41.7% and 62.4%, respectively. The decrease between the second and third periods, and increase between third and fourth periods were statistically significant (chi2 = 4.15, p = 0.04 and chi2 = 8.32, p = 0.004). C. glabrata was the second most common species in the first and second periods (14.8% and 15.5%, respectively), followed by C. tropicalis (12.5% and 10.0%, respectively), however this array has changed in the third and fourth periods (C. tropicalis was the second with the rates of 16.7% and 16.8%, while C. glabrata placed in the third line with the rates of 14.8% and 7.6%, respectively). It was concluded that C. albicans has still been the most frequent species among yeast isolates of ICU's in our hospital; however, the incidence of non-albicans species like C. glabrata and C. tropicalis has increased. PMID- 16358491 TI - [Distribution of Candida species in vaginal specimens and evaluation of CHROMagar Candida medium]. AB - Identification of Candida species is important to guide treatment in vulvovaginal candidiasis which is seen frequently and needs long-term therapy due to recurrence. The aim of this study was to determine the species distribution of Candida isolated from vaginal specimens and evaluation of CHROMagar Candida medium in the laboratory diagnosis. Samples from 80 patients who were clinically diagnosed as vaginitis have been analysed in our laboratory. Colonies appeared on CHROMagar Candida media after 48 hours of incubation at 35 degrees C were evaluated for their colors and characteristics. Candida strains were identified by germ tube test, growth on corn meal Tween 80 agar and when necessary also by API 20 C AUX commercial kit. A total of 84 Candida strains were isolated from 80 patients. Two different Candida species have been isolated from four (5%) of the samples. Among Candida strains isolated, 45 (53.6%) were C. albicans, 29 (34.5%) C. glabrata, 7 (8.3%) C. krusei, and 3 (3.6%) C. kefyr. All of the C. albicans and six of the seven C. krusei isolates have been identified correctly by CHROMagar Candida medium. These results showed that C. albicans is still the most frequently isolated species from vaginal samples. It was concluded that CHROMagar Candida medium is useful for identification of colonies due to frequently seen Candida species and also in differentiation of multiple Candida species grown on the same culture. PMID- 16358492 TI - [Evaluation of three different ELISA kits for the diagnosis of trichinellosis]. AB - Human trichinellosis caused by Trichinella spp. is a widely distributed parasitic disease, acquired by ingestion of undercooked or raw meat containing larvae of the parasite. In January 2004, a trichinellosis outbreak following the consumption of raw minced meat ball, occurred in Izmir, Turkey. In the present study, an in-house excretory/secretory (ES) IgG ELISA and two commercial IgG ELISA kits (Kit # I; IBL, Hamburg, kit # II; Cypress, Belgium) have been used to evaluate their diagnostic efficiencies in this outbreak. Serum samples were collected from 100 patients with trichinellosis, from 16 patients with other parasitosis and autoimmune diseases (2 toxocariasis, 8 hydatic cyst, 1 fascioliasis, 5 rheumatoid arthritis) and from 20 healthy subjects. While the sensitivity of in-house ELISA has been found as 100%, the sensitivities of commercial kits number I and II have been found as 75% and 48%, respectively. The specificities of these three tests have been detected as 93.7%, 87.5% and 100%, respectively. The results obtained in this study suggest that in-house ELISA is of significant diagnostic value for the diagnosis of trichinellosis. PMID- 16358493 TI - [Short communication: clinical predictors of group A beta hemolytic streptococci isolation in upper respiratory tract infections]. AB - The aim of this study was to investigate the clinical prediction criteria for group A beta hemolytic streptococcal (GABHS) pharyngitis in patients with upper respiratory tract infections (URTI). A total of 276 patients admitted to the Ministry of Internal Affairs outpatient clinics with URTI complaints, were included to the study. The clinical findings of the patients were recorded and throat cultures were obtained. The clinical signs and symptoms were as follows; sore throat (56.2%), hypertrophic tonsillitis (53.9%), pharyngeal erythema (50.7%), fever (48.9%), cervical lymphadenopathy (42.8%) and tonsillar exudates (18.5%). The total GABHS isolation rate from throat cultures of the patients were found as 15.9%, and these rates were 19.7% in children, and 7% in adults. Multivariate analysis of the results revealed that the patients with hypertrophic tonsils had GABHS isolation rate seven times more than the patients without hypertrophic tonsils (Odds ratio: 7, confidence interval: 1.4-98, p = 0.017). In conclusion, correct assessment of physical examination findings in the clinical diagnosis of GABHS pharyngitis, will result in less laboratory testing and antibiotic use for sore throat patients. PMID- 16358494 TI - [Short communication: retrospective analysis of 21 HIV/AIDS cases]. AB - In this study, 21 HIV/AIDS cases (18 male, 3 female; age range 17-64 years), followed up in the Department of Infectious Diseases of Uludag University Medical Faculty between 1997-2003 have been analyzed retrospectively, by means of epidemiological, clinical and laboratory aspects. Nineteen (90%) of them were heterosexual, and in 9 cases the diagnosis was coincidental during the blood donations or routine testing. The non-compliance rate of patients to antiretroviral treatment was found as 76%, and the most important factor for non compliance was the difficulty in providing antiretroviral drugs. The most frequently encountered opportunistic infections were oropharyngeal candidiasis (n:5), herpes zoster (n:4) and community acquired pneumonia (n:4). PMID- 16358495 TI - [Short communication: evaluation of the correlation between hepatitis D virus (HDV) RNA positivity and HDV antibodies]. AB - The objective of this study was to evaluate the correlation between serum hepatitis D -delta- virus (HDV) RNA detection and anti-HDV IgG and IgM antibodies, in the serodiagnosis of delta hepatitis. A total of 153 HBsAg positive sera were screened for the presence of anti-HBc IgM, anti-HDV IgG and anti-HDV IgM by commercial enzyme immunoassays and HDV-RNA by real time polymerase chain reaction (PCR). Of 153 sera, 86 (56.2%) were found positive for HDV antibodies. Although isolated anti-HDV IgG was present in 35 and isolated anti-HDV IgM was present in 11 patients, IgG and IgM were present concurrently in 40 additional patients. HDV-RNA was detected in 21.5% (33/153) of the patients. Four of the 33 HDV-RNA positive patients were positive only for anti-HDV IgG, 8 were positive only for anti-HDV IgM, and 19 were positive for both anti-HDV IgG and IgM antibodies. Twenty seven of 51 (53%) anti-HDV IgM positive patients were also found positive for HDV-RNA, while 27 of 33 (82%) HDV-RNA positive patients exhibited anti-HDV IgM positivity. Increased serum ALT levels were detected approximately in 85% (28/33) of viremic patients. As all of the HDV-RNA positive patients were found negative for anti-HBc IgM, superinfection with delta virus were considered. In conclusion, PCR is a sensitive and useful method for the detection of viremic patients as well as for the monitorization of antiviral therapy, anti-HDV IgM positivity together with increased ALT levels appear to be good markers for the prediction of hepatitis delta viremia, especially in the countries with limited economical sources as Turkey. PMID- 16358496 TI - [Case report: a nosocomial infection caused by vancomycin resistant enterococcus]. AB - Vancomycin resistant enterococcus (VRE) was recovered from the urine culture of a 61 years old female patient, who was being treated for sepsis, on the 15th day of hospitalization in Ondokuz Mayis University Hospital Infectious Disease Unit. The underlying diseases of this patient were chronic renal failure and diabetes mellitus. The patient died due to septic shock on the day of VRE isolation. Since this case was the first VRE infection in our hospital, a point prevalence study was planned. For this purpose, rectal swab samples collected from 10 patients from the same unit and 27 personnel who worked in the same unit, were screened for the presence of VRE. Nasal swabs and finger tip samples were also taken from the staff to determine if the transmission has occured in this way. As a result, a second VRE strain was isolated from another patient with chronic renal failure who was under treatment due to multiple pulmonary abscesses. Immediate isolation of this patient prevented a possible epidemic in this specific unit. In this report, the importance of VRE screening and isolation of the patients after the recovery of VRE has been emphasized. PMID- 16358497 TI - [Two leptospirosis cases and review of the national literature]. AB - In this report, two cases of leptospirosis admitted to our clinic in the same periods, have been presented. The first patient was a 29 years old male farmer, and the other was a 44 years old male prisoner. Both of them were from Middle Anatolian Region with the similar clinical findings (subfebrile fever, gastrointestinal complaints, ichterus in sclera, leucocytosis, and increased levels of liver enzymes, urea and creatine). The diagnosis was based on the antibody positivities against Leptospira with microscopic agglutination test which performed in the Etlik Central Veterinary Control and Research Institute. The first case was positive for L. grippotyphosa, and the second was positive for L. australis serovar bratislava at the titers of 1/800. Both of the patients were given empirical antibiotic treatment (ceftriaxone 2 x 1 gr, and ampicillin sulbactam 4 x 1 gr, respectively), and discharged with complete healing. These cases led us to review the other leptospirosis cases in our country. PMID- 16358498 TI - [Immunopathogenesis of Leishmania infections]. AB - Leishmaniasis represents a complex of diseases with a clinical and epidemiological diversity. Leishmaniasis remains a severe public health problem and its burden is increasing. The disease is caused by a parasite belonging to the genus Leishmania. Approximately 350 billion people in 88 different countries are thought to be infected with Leishmania spp. Clinical forms of leishmaniasis are particularly diverse representing different diseases: visceral (VL), cutaneous (CL), diffuse cutaneous (DCL) and muco-cutaneous (MCL) leishmaniasis. Being the most important determinant not only cellular immunity plays the essential role in the control of leishmaniasis, but the virulence, tropism and pathogenicity that is modulated by environmental and genetic factors of their mammalian hosts and sandfly vectors, are the key interactions. These eukaryotic pathogens have evolved with the vertebrate immune system and typically produce long lasting chronic infections. A critical step in their host interaction is the evasion of innate immune defenses. The ability to avoid attack by humoral effector mechanisms, such as complement lysis, and to resist killing by lysosomal enzymes and toxic metabolytes is of particular importance. They do so by remodelling the phagosomal compartments in which they reside and by interfering with signalling pathways that lead to cellular activation. In addition they modify the antigen presenting and immunoregulatory functions of dendritic cells, a process that fascilitates their evasion of both innate and adaptive immunity. Experimental animal studies revealed that these modifications and interference mechanisms led to two different pathogenesis schemes. For CL, the polarization of Th2/Th1 cells is responsible for the progression of the disease which than leads to the chronic-persistant state. The Th2/Th1 paradigm does not apply for visceral leishmaniasis. Immunosupression rather than polarization is responsible for the systemic and progressive outcome of the disease in VL. Based on experience with animal models and humans, new vaccine and novel immunotherapy strategies especially for the locations where the disease is endemic, hold promise for the near future. In this review article the immunopathogenesis of leishmaniasis has been discussed under the light of recent literature. PMID- 16358499 TI - [Viruses as biological weapons]. AB - The destruction made by nuclear, biological and chemical weapons used by governments and terrorist groups in the near history is posing anxiety and fear for human being. Rumour about the possible use of these agents leads to the development of serious negative effects on populations. Since there are no vaccine and therapy for most viral agents and cost of production as biological weapons is low, interest rate is rising for viruses. In this review, general characteristics, diagnosis, therapy and protective measures for viral agents such as variola virus, hemorrhagic fever viruses, encephalitis viruses, Hantaviruses and Nipah viruses, those can be used as biological weapon, have been summarized. PMID- 16358500 TI - [Antibiotic resistance in Helicobacter pylori and molecular methods for the detection of resistance]. AB - Helicobacter pylori which is the major agent causing peptic ulcer, gastric cancer and gastric lymphoma, is identified as a class I carcinogen. Invasive and non invasive tests have been used in the diagnosis of H. pylori infection. Clarithromycin resistance in H. pylori strains is increasing nowadays, thus leading to failures in eradication therapy. Fluorescence in situ hybridization (FISH) method which can simultaneously detect the presence of H. pylori and clarithromycin resistance is a non-culture dependent molecular technique. In this review article the prevalence and clinical outcome of antibiotic resistance in H. pylori, molecular mechanisms of the resistance and molecular methods to detect the resistance, with priority of FISH technique have been discussed. PMID- 16358501 TI - [Agreement of urine culture results with whole urine analysis in patients with urinary tract infections]. PMID- 16358502 TI - Science-based assessment of animal welfare: farm animals. AB - Animal welfare is to do with the feelings experienced by animals: the absence of strong negative feelings, usually called suffering, and (probably) the presence of positive feelings, usually called pleasure. In any assessment of welfare, it is these feelings that should be assessed. Because feelings are subjective, they cannot be investigated directly. However, there are indirect methods by which animals can be 'asked' what they feel about the conditions under which they are kept and the procedures to which they are subjected. These methods involve preference tests, followed up by motivational tests to assess how important the animal's choice is. Measurements of impaired biological functioning, particularly those connected to decreased health and increased physiological stress responses, can provide good corroborating evidence that welfare is compromised. PMID- 16358503 TI - Science-based assessment of animal welfare: companion animals. AB - Human history reveals that the way in which humans treat animals is based on their views of themselves as well as of the living environment around them. These views may vary from an assumption of human superiority to one of equality between humans and animals. Recent trends affecting companion-animal welfare are: modern philosophies on animal issues, the specialised and varied roles that companion animals play in modern societies, new results from animal neuroscience, human animal interaction studies and the new profession of companion animal ethology. This paper concludes that applied ethology could provide science-based criteria to assess companion-animal welfare. Due to the integral part that companion animals play in human societies, the paper is divided into an animal component that deals with the animal's basic needs and its ability to adapt, and a human component assessing the living environment of animals as provided by humans and responsible companion-animal ownership. The greatest challenge for future research is to find ways to disseminate knowledge of companion animal ethology to companion animal owners. PMID- 16358504 TI - Science-based assessment of animal welfare: laboratory animals. AB - The use of animals in experimental research parallels the development of medicine, which had its roots in ancient Greece. The increasing demand for high standard animal models, together with a critical view of the way animals are used, has led to the development of a multidisciplinary branch of science we now know as 'laboratory animal science'. The guiding principles are replacement, reduction and refinement (the Three Rs), first proposed by Russell and Burch in 1959. When animals are used, the people involved have an obligation to safeguard their welfare and minimise discomfort; this will also generally be beneficial for both the animal and the experimental outcome. The ability of an animal to cope with the environment and exert control over its life seems to be crucial for animal welfare. In this paper, attention is paid to the assessment of welfare, environmental factors affecting welfare, legislative requirements and future trends such as the production and use of genetically modified animals. PMID- 16358505 TI - Science-based assessment of animal welfare: wild and captive animals. AB - Science-based assessment of animal welfare in wild and captive animals has, during the past ten years, benefited from increased interest in and research into consciousness, emotions, stress and pain. Because it is impossible to make a detailed physiological study in free-living animals, reliance must be placed on behaviour and Darwinian fitness, which have been shown to correlate to welfare. Only in captive animals can more detailed physiological studies be made, and then not without difficulty. In short, the welfare of an animal is shown to be compromised if the animal exhibits similar behaviour to that of animals whose physiological responses have been confirmed as indicating poor welfare, and if its Darwinian fitness is compromised in similar ways to such animals. PMID- 16358506 TI - Science-based assessment of welfare: aquatic animals. AB - Terrestrial animal welfare has been a matter for exploration for many years. In contrast, approaches towards improving the welfare and humane treatment of aquatic animals are relatively new, as is the thinking behind them. Several issues complicate the process of addressing the welfare of aquatic animals in a consistent manner. These include the following: - the huge diversity among aquatic animals, the majority of which are poikilothermic vertebrates and invertebrates - understanding the practices involved in fisheries, aquaculture and aquatic animal production, and their purpose - the relative paucity of scientific information - understanding the philosophical approaches, policies, guidance and regulations that may influence the provision of optimal welfare and humane practices for aquatic animals. In this paper, the authors attempt to provide an overview of all these elements, relating what is known and understood about these issues for the primary group used in aquaculture and fisheries, finfish, and exploring the factors that may influence the concepts and practices of aquatic animal welfare. These factors seem to be the foundation of all welfare approaches and include: - ethical and moral concepts of animal welfare and humane treatment - whether animals experience suffering from the potentially adverse practices used in their maintenance, management and use - the public and institutional understandings of these issues and their results. These are discussed with the hope that future developments in, and approaches to, aquatic animal welfare will be of use to society, industries and the public. PMID- 16358507 TI - [Animal health policies and practices in the Americas: preliminary study]. AB - The Americas have a large population of farm animals, mostly for export. There are diverse production systems distributed over an extensive and varied geography, which hampers efforts to respond to the demands of the different markets. This study provides an overview of the elements influencing animal welfare implementation, such as the requirements of importing countries, the requirements of private agents, the demands of producers and manufacturers, quality promotion policies, the demands of the community, the recommendations of reference bodies and the results of applied research. To explore the level of animal welfare development in the countries of the region, a detailed case study was made of Chile, in addition to a survey of the Member Countries of the World Organisation for Animal Health (OIE) in the Americas. An analysis was made of progress with the issues considered by the OIE as priorities, namely humane slaughter for human consumption, transport and killing for disease control purposes. Furthermore, the study considers various aspects of production which the OIE has not included up to now. It also explores the status of research and producer and consumer perceptions of the issue. The results reveal that the level of development and implementation of animal welfare differs from one country to another. While the adoption of animal welfare regulations certainly relates to all the above-mentioned aspects, the one which appears to have the most impact is the export of livestock products to certain markets. Although there is great interest in improving animal welfare conditions, this calls for the general characteristics of animal husbandry in the various countries to be taken into account. While some livestock production in the Americas follows world patterns, many countries still find it difficult to integrate good animal welfare practices, owing to specific geographical, social and cultural situations that are reflected in local livestock development rationales. PMID- 16358508 TI - Global perspectives on animal welfare: Europe. AB - Effective implementation and enforcement of legislation is essential to ensure animal welfare. In the World Organisation for Animal Health (OIE) European Region the well-established body of national and European Union laws in existence is growing continuously. The growth is due to various factors, such as new technology in animal farming and experimentation, exploitation of wildlife, new understanding of animal needs, and increasing public awareness and concern. The latter, in particular, determines the need for new animal welfare legislation to regulate and discipline the 'use' of animals for different purposes, such as food production, companionship, work and leisure. This paper intends to provide an overview of the more relevant activities carried out by the Council of Europe and the European Union in the field of animal welfare. The authors identify eLearning as a tool to harmonise the interpretation and the implementation of animal welfare legislation. PMID- 16358509 TI - Global perspectives on animal welfare: Africa. AB - Livestock production systems, production objectives, the cultural values of livestock keepers, and the close relationship between keepers and their livestock have evolved over the years and have influenced the quality of animal welfare in Africa. An equivalent level and quality of care is not evident for companion animals, especially dogs and donkeys, who are often mistreated and physically abused. In the densely populated highland and humid coastal belts of Africa, profit-driven commercial large-scale intensive livestock production systems predominate. As the main production objective of these operations is to maximise profit, the operators of these production systems do not exhibit the same kind of attachment to their livestock as traditional farmers. In some large-scale commercial systems animals and birds are kept in sub-standard poorly constructed structures that greatly restrict the animals' movements. In Africa, conservation of wildlife habitats is part of animal welfare, but due to an increasing human population and a greater demand for land for grazing, cultivation, and housing, wildlife reserves are quickly diminishing. This essentially means that the land that was previously set aside for wildlife and forests is being encroached upon and previously unsettled plains and marginal lands are being used for agriculture and mining. In most places there is significant conflict between humans and wildlife, such that wildlife are considered to be pests that need to be destroyed. This is a particular problem in areas where wildlife have destroyed crops, attacked man and/or livestock, or there has been disease transmission between animals and humans. In situations and/or areas that do not have official animal control services, crude weapons, poisons, and traps are used to kill the wildlife. Animal welfare issues, domestic and wildlife related, need to be urgently addressed through policy and legal frameworks and supported by community awareness of, education about, and participation in, animal welfare issues. PMID- 16358510 TI - Global perspectives--the Middle East: Egypt. AB - The Middle East region is characterised by the grouping of 20 countries with similar cultural traditions and religious concepts. The majority of the population of this region follows the religion of Islam, which considers all animals as important as humans, and others follow Christianity and Judaism. The majority of people dealing with animals have their own religious concepts, but most countries have no clear enforced official animal welfare regulations. Animal welfare activities in the Middle East region, as in Egypt for example, are carried out by various individuals, non-governmental organisations and the official Veterinary Services. The slaughter of animals for human consumption is in accordance with Islamic rites, which is the least painful method for the animals. Extensive production systems are the predominant systems in the region, and it is not common for animals in the region to suffer stress during production processes or slaughter. Although some success has been achieved in the area of animal welfare, as seen in improvements in health care, the development of modern abattoirs with proper facilities, and the establishment of policies for combating stray dogs, there are still major constraints to the implementation of effective animal welfare measures, including those for wildlife. These constraints include the lack of finances and the absence of effective legislative measures to prevent the trapping and/or hunting of wild animals for pleasure or illegal trade. PMID- 16358511 TI - Global perspectives on animal welfare: Asia, the Far East, and Oceania. AB - In Asia and the Far East, livestock undergo major suffering due to malnutrition, overloading, and ill-treatment. At slaughter animals are handled roughly and watch other animals being killed; stunning is not practised. Cruelty to other animals such as elephants, horses, donkeys, bears, dogs, and circus animals has largely been prevented through the efforts of animal welfare organisations. Governments have taken initiatives to establish Animal Welfare Boards and enact laws for the prevention of cruelty to animals, but their efforts are far too limited to be of any significance and financial constraints and lack of personnel inhibit the implementation of the laws that do exist. In New Zealand and Australia, legislation and strong consultation procedures at governmental and community level strive to regulate and improve the welfare of animals in all spheres, but in other Oceanic countries there is a need for both an update in, or establishment of, legislation covering animal welfare. Limited progress has been made due to the status of the Veterinary Services and a lack of resources. Although some public and educational awareness programmes are carried out, increasing exposure to international media and attitudes of visiting tourists suggest that further awareness work needs to be undertaken. To address the problems of animal welfare in developing countries, it would be inappropriate to adopt the international standards that are implemented in the developed countries. Each developing country should evolve its own standards based on its own individual priorities. PMID- 16358512 TI - Animal welfare: the role and perspectives of the meat and livestock sector. AB - Those in the livestock industry involved in rearing animals and in producing milk, meat and eggs, must respond to two demands: one expressed by consumers, and the other by the public. Regarding consumers, demand for food produced by the livestock industry has shown steady growth for a century. Over the last few decades, this growth has been sustained by the developing countries, and is based mainly on pig and poultry production, which provides cheaper products. Regarding the public, society is showing greater concern about the conditions in which livestock is reared, transported and slaughtered. The public demands not only that ill treatment of animals be fought against and penalised, but also that any unnecessary suffering should be avoided and even that animals should be guaranteed a certain degree of 'comfort'. Animal health, the most important aspect of their welfare, has vastly improved, as has the care of sick or injured animals. At the same time, the latest amenities used in livestock rearing, transport and slaughter are helping to eliminate situations involving extreme stress and suffering. Finally, the motivation of industry players and the safety of those who work with livestock must be taken into consideration. Training of personnel and the implementation of guides to good practice or of quality control do as much to improve animal welfare as do overzealous regulations. PMID- 16358513 TI - Animal welfare: the role of non-governmental organisations. AB - The welfare of animals is of interest to many people in most parts of the world. Concern about the way that animals are treated will depend on many factors, including socio-economic conditions, culture, religion and tradition. The World Organisation for Animal Health (OIE) is committed to ensuring that all animal welfare standards are science-based, but recognises that these other factors must also be taken into account. The International Coalition for Farm Animal Welfare (ICFAW) was formed to represent the interests of non-governmental animal welfare organisations from most corners of the globe and opinions, comment and information from these animal welfare organisations will play a part in the OIE decision-making process. In coming together for this purpose it was recognised that the views of the various member organisations of ICFAW vary depending on which part of the world they come from. The authors provide information about the situation in three continents: Africa, North America and Europe. This information includes details of relevant legislation, farming practices, and educational and campaign programmes developed by both animal welfare non-governmental organisations and governments. The authors also look to the future to see what issues may influence the way that farm animals are reared, transported and slaughtered. PMID- 16358514 TI - The World Veterinary Association and animal welfare. AB - The World Veterinary Association, as the global representative of the veterinary profession, recognises the global influences on animal welfare and the changing role of the veterinarian in response to the changing attitudes of the human population. While urban populations are now dictating animal welfare standards, many practices still have a cultural and even religious basis. Veterinarians recognise these influences, but base their recommendations for animal welfare on scientifically justified practices. Veterinarians work not only for urban clients with their companion animals, but also very importantly with rural clients who provide the source of animal-based foodstuffs and goods sought by an increasingly demanding human population. The controversial areas of intensive animal production and the transportation that is required to move large numbers of animals around the world require veterinary supervision to ensure that animal welfare is preserved. The development of animal welfare standards is an ongoing process, with the major international effort being led by the World Organisation for Animal Health. PMID- 16358515 TI - Animal transport and welfare: a global challenge. AB - Animal welfare has moved on from being a concern of a caring few to the stuff of international legislation. The author argues that more research is needed into animal welfare, especially since legislation should be based on sound science rather than emotion or anthropomorphism. The farming and animal handling community should consider the potential benefits of animal welfare rather than viewing the whole issue as a threat. The animal welfare community can assist both animals and themselves by stressing the economic advantages to be obtained from better animal welfare. Better welfare requires more than just being kind to animals, but a combination of skills, science and experience that need to be combined into one overall effective discipline. PMID- 16358516 TI - The Food Marketing Institute and the National Council of Chain Restaurants: animal welfare and the retail food industry in the United States of America. AB - In order to achieve real change, there must be a motivating force and all the stakeholders need to be involved. This is the premise of the animal welfare programme developed for the food retail, wholesale and chain restaurant industries in the United States of America (USA) by the Food Marketing Institute (FMI) and the National Council of Chain Restaurants (NCCR). This paper outlines a collaborative process that retailers and producers in the USA are using to enhance the care and welfare of animals in commercial food production. Although the efforts of the FMI and the NCCR are still underway, the process provides one example of how different parts of the food production system can work together to achieve positive change. PMID- 16358517 TI - Philosophy, policy and procedures of the World Organisation for Animal Health for the development of standards in animal welfare. AB - Animal welfare was identified as a priority for the World Organisation for Animal Health (OIE) in the 2001-2005 OIE Strategic Plan. Member Countries recognised that, as animal protection is a complex, multi-faceted public policy issue which includes important scientific, ethical, economic and political dimensions, the OIE needed to develop a detailed vision and strategy incorporating and balancing these dimensions. A permanent working group on animal welfare was established in order to provide guidance to the OIE in its work on the development of science based standards and guidelines. The Working Group decided to give priority to the welfare of animals used in agriculture and aquaculture, and that, within those groups, the topics of transportation, slaughter for human consumption and killing for disease control purposes would be addressed first. Some guiding principles were approved by the International Committee of OIE Member Countries during the 72nd General Session in May 2004, and these have been followed by four specific guidelines on the priority topics listed above. PMID- 16358518 TI - Transport of animals by sea. AB - The author briefly describes the development of the World Organisation for Animal Health (OIE) Guidelines for the transport of animals by sea. The paper provides details about the membership of the OIE Ad hoc Group on the sea transport of animals, the terms of reference of this group, the process followed and the consultations that were undertaken in developing and reviewing the guidelines. An outline of the guideline contents is also provided. In addition, the author outlines the programme of research undertaken into the causes of sheep and cattle deaths during sea transport from Australia--the world's largest exporter of sheep and cattle by sea. The main findings of these investigations are summarised. PMID- 16358519 TI - The effects of land transport on animal welfare. AB - Animal welfare during and as a result of transport can be assessed by using a range of behavioural, physiological, pathological and carcass-quality indicators that are described in this paper. Measures of the extent of any disease, injury or mortality resulting from, or exacerbated by, transport are important because health is an important part of welfare. Many of the indicators are measures of stress as they involve long-term adverse effects on the individual. Factors affecting the welfare of animals before, during and after transport which are discussed are: definition of the responsibilities and competence, attitudes to animals and need for training of staff; planning of journeys and methods of payment of staff; laws and retailers' codes; genetics, especially selection for high productivity; rearing conditions and experience; the mixing of animals from different social groups; handling and loading procedures; driving methods; space allowance; increased susceptibility to disease and efforts to minimise the spread of disease. PMID- 16358520 TI - Slaughter of animals for human consumption. AB - This paper briefly describes the formation of the OIE (World Organisation for Animal Health) Ad hoc Group on Humane Slaughter of Animals, summarises its initial discussions and provides details of its recommendations. These recommendations were approved (with some amendments) at the 73rd General Session of the OIE International Committee and have been included in the 14th edition of the OIE Terrestrial Animal Health Code (2005). The recommendations include some changes to current definitions of certain terminology, and guidelines pertaining to the commercial slaughter of animals for human consumption, designed to minimise avoidable pain and suffering at every stage of the pre-slaughter and slaughter processes. The main issues addressed by the Ad hoc Group were: general principles for slaughter, as related to personnel, animal behaviour, and the removal of distractions which can cause animals to stop or turn back; moving and handling animals following their arrival at the slaughterhouse; lairage design and construction; care of animals in lairages; and animal welfare issues associated with acceptable handling, restraining, stunning and slaughter methods applicable to various relevant species. The group acknowledged the significance of religious requirements as well as the cultural and ethnic factors associated with slaughter, and appropriate proposals were included in their final report. Important but less frequently occurring issues such as the management of foetuses during the slaughter of pregnant animals were also included in this work. Finally, the report of the Ad hoc Group also specified methods, procedures and practices that are unacceptable on animal welfare grounds. PMID- 16358521 TI - Killing of animals for disease control purposes. AB - In emergency disease outbreaks, large numbers of animals may need to be killed for control, containment and/or eradication purposes. Strategies for disease control will usually involve complete depopulation of herds/flocks, and may extend to depopulation across designated areas. Live animals present the major risk of spreading infectious agents so their slaughter should be completed as quickly as possible, with minimal pain and distress to the animals. The method chosen for killing of animals must be humane, efficient and safe for people and the environment. The tasks should be conducted under the supervision of an official veterinarian, supported by a sufficient number of adequately trained and competent personnel. It is essential to ensure all animals are dead, as demonstrated by the cessation of cardiac and respiratory movements. This paper discusses methods by which animals may be killed for disease control purposes. PMID- 16358522 TI - The assessment and implementation of animal welfare: theory into practice. AB - This paper reviews the procedures and protocols necessary for the development of a practical programme for the assessment and implementation of animal welfare on farms and in other commercial situations. An effective programme must incorporate measures of both husbandry and welfare. Most current systems are based almost entirely on measures of husbandry provision, e.g. resources and management. However, what the public and the animals need are guarantees of satisfactory welfare and these guarantees must involve animal-based measurements of welfare outcomes. The development and testing of the Bristol Welfare Assurance Programme is described in outline and illustrated with detailed reference to the assessment of welfare in dairy cows. The final section describes how such a programme can be incorporated into the broader concept of quality assurance schemes that incorporate a proper concern for animal welfare into the reasonable needs of different societies for wholesome, affordable food. PMID- 16358523 TI - Using animals in research, testing and teaching. AB - The authors describe landmarks in animal-based research and examine key moral statements on the use of animals in scientific research. The principles of 'reduction, replacement and refinement' are suggested as a focus around which both the animal welfare and the scientific communities of the world can unite. This paper reviews the progress achieved in the replacement of animal use by non animal methods, with a focus on regulatory testing. In addition, it presents evidence of a reduction in the numbers of animals used throughout the 1990s. The authors also emphasise the need for concerted efforts to contain a potential escalation in these figures, resulting from current inefficiencies in creating genetically modified animals and increasing demands for regulatory testing. Finally, the authors examine the refinement of techniques to mitigate and prevent pain and distress, with an emphasis on appropriate endpoints as an effective tool. They present the need for the international harmonisation of ethical standards and processes, together with a suggested harmonisation platform, and demonstrate the central role that should be played by the institutional Animal Care Committees. PMID- 16358524 TI - Animal welfare and international trade. AB - Globalisation is becoming a force that is revolutionising international trade, particularly that of animals and animal products. There is increasing interest in animal welfare worldwide, and as part of its 2001-2005 Strategic Plan the World Organisation for Animal Health (OIE) identified the development of international standards on animal welfare as a priority. The OIE's scientific approach to standard-setting provides the foundation for the development, and acceptance by all OIE Member Countries, of these animal welfare guidelines. The paper discusses how these guidelines on animal welfare can be implemented, both within the provisions of World Trade Organization (WTO) agreements and within the framework of voluntary codes of conduct. Even if animal welfare guidelines are not covered by any WTO agreements in the future, bi- and multilateral agreements, voluntary corporate codes, and transparent labelling of products should result in a progressive acceptance of OIE guidelines. Ultimately, consumer demands and demonstrable gains in animal production will result in an incremental evolution in animal welfare consciousness and adherence to international standards. PMID- 16358525 TI - Food safety and environmental issues in animal welfare. AB - Food safety, protection of the environment and animal welfare are three concerns consumers have about modern food production systems. Standards dealing with these three issues can be in conflict, and finding ways to reduce this conflict is essential as international standards governing animal welfare develop. These conflicts can be reduced if flexible animal welfare standards are animal-based and attempt to directly assess the state of the animals themselves rather than prescribing the conditions under which the animals are reared. Animal welfare is often treated as an ethical issue, and is therefore difficult to deal with under current trade agreements. A greater appreciation of the link between animal welfare and animal health makes the link with food safety clearer. Improvements in animal welfare have the potential to reduce on-farm risks to food safety, principally through reduced stress-induced immunosuppression, reduced incidence of infectious disease on farms and reduced shedding of human pathogens by farm animals, and through reduced antibiotic use and antibiotic resistance. Health problems of farm animals continue to be serious threats to animal welfare, and measures of disease incidence can serve as animal-based measures of animal welfare. Continued development of hazard analysis and critical control point based approaches to animal welfare would allow a smoother integration of animal welfare and food safety standards. PMID- 16358526 TI - Strategic animal welfare issues: ethical and animal welfare issues arising from the killing of wildlife for disease control and environmental reasons. AB - Ethical and animal welfare concerns about the destruction of free-living wildlife for disease control and environmental reasons have historically received little attention from animal welfare scientists, legislators or the public. Nevertheless, all vertebrates can experience pain and distress, regardless of whether they are unwanted pests or not. A wide range of methods is used to kill or otherwise control unwanted wildlife. The animal welfare impacts of most of these methods are not known. The extremely high number of animals being controlled and the potential impacts of this control on their welfare suggest that this is a significant strategic issue that should be considered by the World Organisation for Animal Health (OIE). This is particularly important, given the focus of the OIE on disease control and the requirement for pest control as a part of this process. PMID- 16358527 TI - Animal welfare and developing countries: opportunities for trade in high-welfare products from developing countries. AB - Discussion on the potential for developing countries to develop trade in niche markets such as higher welfare standards has been highlighted with moves by the World Organisation for Animal Health (OIE) to set internationally agreed standards for animal welfare. This paper examines the existing and potential trade in value-added higher welfare products using case studies in the beef and poultry sectors from three countries in Africa, Asia and Latin America. It shows that at present there is only a small trade in these products but that this can have a major effect at a national level. In the beef export trade from Namibia, the existence of the only assurance scheme in Africa setting standards in hygiene, veterinary care and animal welfare has created a trusted, safe and healthy product and ensured that Namibia has grown into Africa's largest exporter of beef to the European Union. In Thailand, the broiler industry, which has enjoyed annual growth in the past 15 years, is developing value-added products to develop markets to counter competition from other countries. The development and implementation of standards for organic products in both Thailand and Argentina over the past decade have also resulted in growth in the export markets of these products. The paper concludes that there is growth potential for the sectors in all three markets which can be assisted by the development of OIE baseline standards. PMID- 16358528 TI - The use of animals in agriculture and science: historical context, international considerations and future direction. AB - As the final contribution to this important World Organisation for Animal Health (OIE) publication, this paper provides some relevant background and contextual information and identifies a number of strategically significant international activities that will influence the future direction of animal welfare internationally. The assumption of an animal welfare leadership role by the OIE, with the full support of its 167 Member Countries, is an international development of major strategic significance. As an inter-governmental organisation, the OIE is committed to a science-based approach to the development of animal welfare guidelines and standards and to working closely with all stakeholders. This paper covers the use of animals in both agriculture and science, reflecting the OIE's dual remit for both animal health and animal welfare and the importance of animal-based research and testing to the OIE's animal health and reference laboratory roles. PMID- 16358529 TI - A conversation with Bill Borne, CEO, Amedisys, Inc. Interview by Val J. Halamandaris. PMID- 16358530 TI - A journey to excellence in care. PMID- 16358531 TI - Finding the heart of home care: the Bayada Way project. PMID- 16358532 TI - The future of hospital based homecare is in a free standing structure! BJC health care has proof--part II. PMID- 16358533 TI - Six Sigma: customer driven quality improvement with the bottom line in mind. PMID- 16358534 TI - The data is in: best home care marketing practices revealed. AB - The first in a two-part series, this article provides an early look at a study evaluating the best marketing practices of home health agencies across the country. In Part 1, we profile a few exemplary agencies with the goal of describing a set of common attributes that have lead to success. In Part 2 (January), we will delve deeper into the component measures by exploring the first industry-specific benchmarks of marketing success! PMID- 16358535 TI - A guide to workers' compensation nirvana. PMID- 16358537 TI - Top 10 ways to have a successful Home Health & Hospice agency. PMID- 16358536 TI - The National Quality Improvement/Hospitalization Reduction Study, Part I. An industry-wide response. AB - It is the one Home Health Compare measures that has not improved in three years. It has been a major area of concern for most home care agencies in the country. It is THE focus of QIOs quality improvement efforts. And, it has serious implications to agencies quality status in a pay-for-performance environment. PMID- 16358538 TI - Survey & certification Q & A on the Medicare Hospice Benefit from the CMS Hospice Expert Panel. PMID- 16358539 TI - Retiring baby boomers create opportunities for innovation. PMID- 16358540 TI - Where is your tipping point? PMID- 16358541 TI - Q & A you ask and Sam Geist answers. Personnel selection. PMID- 16358542 TI - Why you may have a rooting interest in publicly traded competitors. PMID- 16358543 TI - Is your disaster plan up to date? PMID- 16358544 TI - The Huichol experience: healing wounds. PMID- 16358545 TI - A message to the missionaries. PMID- 16358546 TI - Moving the New York State Research Agenda forward: the stakeholder is you! AB - Health care is more diverse than it has ever been. The point of care can be rural or urban, inpatient or outpatient, individual or community-focused. In this challenging, ever-changing environment, it is critical for nurses to have access to nursing research on the best way to provide care. The New York State Nursing Research Agenda was launched in 2001 as an ongoing action plan to facilitate the conduct, dissemination, and utilization of nursing research in New York. This article describes several initiatives that have been launched to implement that plan. Strategies for local and regional initiatives are outlined. Key to the success of these initiatives is the involvement of nurses at all levels of practice and in all healthcare arenas. All nurses are stakeholders because this is our science and our profession. PMID- 16358547 TI - The development of nursing research self-study modules. AB - Research utilization and evidence-based practice are processes that require nurses to have research knowledge and skills. Yet many nurses are fearful of research and avoid reading nursing research studies or using research in their practice. Members of a hospital's Nursing Research Team developed nursing research self-study modules for use by the nursing staff. The goal of this innovative project was to enhance a nurse's ability to critically read nursing research studies and determine their usefulness for practice. As a result, it is hoped that nurses will be more knowledgeable about research and more comfortable participating in the evidence-based practice and research utilization processes. The development, implementation, and evaluation of nursing research self-study modules are described and implications for nursing practice identified. PMID- 16358548 TI - Critical care nurses' experiences when technology malfunctions. AB - When caring for critically ill patients, critical care nurses work with technology every day. Technology and equipment malfunctions can have a profound effect on nurses' practice and self-image. In this article, a descriptive phenomenological methodology was chosen to explicate the experience of seven critical care nurses. While participants realized that machines might malfunction, they experienced surprise, shock, and feelings of being "let down" and inadequate when malfunctions occurred. They questioned their competence and felt malfunctioning technology jeopardized their credibility and professional image. These findings are useful when structuring educational sessions on technology and in facilitating a supportive environment for critical care nurses when technology malfunctions. PMID- 16358549 TI - Partnership model for practice and education. AB - The faculty practice partnership model provides a framework for collaboration between a practice setting and a university school of nursing. The model created between the Bergen County Department of Heath Services (BCDHS), the Office of Public Health Nursing (PHN) and Pace University's Lienhard School of Nursing (LSN) is one that supports faculty practice and student involvement in population based activities. The partnership provides BCDHS with the assistance it needs to provide the three core functions of public health: assessment, assurance, and policy development. It provides LSN with a practice site where faculty members are able to maintain their clinical expertise, as well as a site for student clinical experiences and research. This article describes the partnership model and its accomplishments from both practice and educational perspectives. PMID- 16358550 TI - Efficacy of medium-chain triglycerides compared with long-chain triglycerides in total parenteral nutrition in patients with digestive tract cancer undergoing surgery. AB - The objectives of this prospective, randomized study were to evaluate the efficacy and tolerability of the short-term use of medium-chain triglyceride/long chain triglyceride (MCT/LCT) fat emulsions, and to compare the hematologic and biochemical effects of MCT/LCT fat emulsions with LCT fat emulsions in gastrointestinal (GI) tract cancer patients following surgery. Thirty patients with GI tract cancer requiring total parenteral nutrition (TPN) were equally randomized to receive MCT/LCT or LCT emulsions for 7 days. After 7 days, no sign of complications directly related to administration of fat emulsions was observed and there were no marked differences in anthropometry, length of hospital stay, and surgical complication rates between the two groups. However, MCT/LCT significantly improved plasma prealbumin concentration (p = 0.005). Changes in complements C3 and C4, total lymphocyte count, and immunoglobulins after TPN were not significantly different between the groups. Serum triglyceride and cholesterol levels remained constant. The serum insulin level in the MCT/LCT group was higher than in the LCT group (p = 0.048). Our data revealed that MCT/LCT fat emulsions significantly enhanced nutritional status in patients with GI tract cancer, indicated by higher prealbumin levels, which might be partially due to the higher circulating insulin levels in the MCT/LCT group. PMID- 16358551 TI - Association between cigarette smoking and hypoxanthine guanine phosphoribosyltransferase activity. AB - The aim of this study was to investigate the association between smoking behavior and hypoxanthine guanine phosphoribosyltransferase (HGPRT) activity. A cross sectional study was performed of 82 men, including 38 non-smokers and 44 smokers. Inosine monophosphate (IMP), the product of HGPRT (used as the index of activity), was measured in peripheral blood mononuclear cells using high performance liquid chromatography. The factors potentially associated with HGPRT activity included age, glutamyl oxaloacetic transaminase, glutamyl pyruvic transaminase, cholesterol, uric acid, triglycerides, creatinine, body mass index, gout, systolic blood pressure, diastolic blood pressure, alcohol consumption, and cigarette smoking. Mean HGPRT activity was 7.05 +/- 3.44 nmol/10(6) viable cells/hour for all participants, and was significantly lower for smokers than for non-smokers (6.24 +/- 3.40 vs 7.98 +/- 3.28 nmol/10(6) viable cells/hour; p = 0.02). In addition, as the number of smoked cigarettes increased, the HGPRT activity decreased (p < 0.05). The age at onset of cigarette smoking showed a positive correlation with HGPRT activity after adjusting for smoking duration, serum uric acid, and cigarettes smoked per year using a multiple regression model (p < 0.001). We concluded that the greater the number of cigarettes smoked, the lower the HGPRT activity, and that HGPRT activity was higher in smokers who had started smoking later. PMID- 16358552 TI - Maternal demographic and psyhosocial factors associated with low birth weight in eastern Taiwan. AB - The relationship between birth weight and maternal sociodemographic characteristics was examined in a sample from two teaching hospitals in eastern Taiwan. Using a structured questionnaire, we conducted face-to-face interviews with women at antenatal clinics between 1998 and 1999 in Hualien City. One year later, we took the outcome of pregnancy from medical records and birth certificates from the Public Health Bureau of Hualien County. Of the 1,128 single live births, 6.8% had low birth weight (LBW) using the World Health Organization cut-off of 2,500 g. LBW was more common in teenage (< 20 years), older (> 30 years), first-time, and unmarried mothers; those with basic/intermediate educational attainment; and residents of aboriginal districts. Teenage pregnancies were more likely than those in adults to be unplanned, and such mothers had smoking or alcohol-drinking behavior. Prevention of teenage pregnancy is crucial to lower LBW rates in eastern Taiwan. For adult mothers, basic or intermediate educational attainment, residence in an aboriginal district, and first-term pregnancy were significant factors associated with LBW, after adjustment for other psychosocial attributes, such as psychologic distress and poor family support. Thus, we should pay more attention when caring for pregnant women with such sociodemographic characteristics, and ensure that they have adequate prenatal care and can adopt a healthy lifestyle. PMID- 16358553 TI - Tolerability of ramipril 10 mg daily in high-risk cardiovascular patients in Taiwan: experience from Kaohsiung Medical University Chung-Ho Memorial Hospital. AB - The Heart Outcomes Prevention Evaluation (HOPE) study demonstrated that the angiotensin-converting enzyme inhibitor, ramipril, significantly reduces mortality, myocardial infarction and stroke in high-risk cardiovascular patients, beyond the benefits from blood pressure lowering. The tolerability of ramipril 10 mg/day has been an important concern when applying these results. Following the same criteria as the HOPE study, we investigated the adverse effects profile and tolerability of 10 mg ramipril in high-risk patients at our institution. In total, 92 patients with high cardiovascular risk were eligible for this study. Initially, ramipril was prescribed 2.5 mg orally once daily, and then titrated up to 5.0, 7.5, and 10.0 mg/day at 1-month intervals. The target maintenance dose was 10 mg/day. All adverse events were recorded during at least 3 months of follow-up. After 4-6 months of the titration protocol, only 18 patients (25.3%) reached and remained on ramipril 10 mg/day; 11 (15.5%), 22 (30.9%), and 20 patients (28.2%) remained on 2.5, 5.0, and 7.5 mg/day, respectively. Twenty-one patients (22.6%) had at least one adverse event. Twelve patients (13.0%) stopped treatment because of adverse effects. A total of 23 episodes of adverse events were reported, including cough (15.1%), dizziness (6.0%), and hypotension (2.4%). Ramipril was relatively well tolerated in our study population. However, only one quarter of our patients reached the target maintenance dose of 10 mg/day. Dry cough, dizziness, and hypotension were the major side effects. About 15% of our patients discontinued ramipril treatment, which is comparable with previous reports. PMID- 16358554 TI - Ruptured aneurysm of the sinus of valsalva into the right atrium without ventricular septal defect: a case report and literature review. AB - Aneurysm of the sinus of Valsalva (ASV), frequently associated with ventricular septal defect (VSD), is a rare cardiac disease that may be acquired or congenital. Rupture of an ASV, rare in the noncoronary cusp, usually produces serious hemodynamic change and carries poor prognosis if not treated surgically. We present the case of a 55-year-old female who came to us complaining of exertional dyspnea. Transthoracic echocardiography and aortography showed a noncoronary cusp ASV with rupture into the right atrium but without VSD. Because of high left to right shunt flow, she underwent successful surgical intervention with aneurysm repair approached from both the aorta and right atrium with a knitted Dacron patch. This was a rare case of noncoronary cusp involvement in ASV that ruptured into the right atrium without VSD. PMID- 16358555 TI - Diffuse abdominal uptake mimicking peritonitis in gallium inflammatory scan: an unusual feature of acute Q fever. AB - The clinical features in patients with acute Q fever are variable. We present a patient with fever, abdominal distension, pericardial effusion, and diffuse gallium uptake in the abdominal cavity, mimicking peritonitis or peritoneum carcinomatosis. Serologic surveys revealed acute infection by Coxiella burnetii. The patient responded poorly to doxycycline and improved with oral levofloxacin. During the afebrile period, gallium inflammatory scan showed resolution of previous diffuse uptake in the abdomen, and cardiac echo resolution of pericardial effusion, which was suggestive of peritoneal inflammation related to acute C. burnetii infection. Therefore, clinicians in Taiwan should be alert to the possibility of acute Q fever in patients with fever of unknown cause, especially with clinical evidence of peritoneal and/or pericardial inflammation. PMID- 16358556 TI - Successful treatment of cerebral venous thrombosis associated with bilateral internal jugular vein stenosis using direct thrombolysis and stenting: a case report. AB - Cerebral venous thrombosis (CVT) is not uncommon, but CVT associated with bilateral internal jugular vein stenosis (BIJVS) is rare. Bilateral internal jugular vein stenting is also a rare procedure. We report on a patient with CVT associated with BIJVS, who was treated successfully using direct thrombolysis and bilateral internal jugular vein stenting. PMID- 16358557 TI - Delayed postpartum hemorrhage--a rare clinical presentation of thrombotic thrombocytopenic purpura-hemolytic uremic syndrome: a case report. AB - Thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS) are rare and closely related disorders characterized by microangiopathic hemolytic anemia, thrombocytopenia, and renal failure. Many risk factors have been reported including infection, cancer, pregnancy, a variety of drugs (e.g. anticancer drugs), and autoimmune diseases. The incidence of TTP-HUS is higher in females than in males, especially during pregnancy and the immediate postpartum period. Review of the literature reveals that delayed postpartum hemorrhage is a rare clinical presentation of TTP-HUS. We report a case of TTP-HUS with recurrent delayed postpartum hemorrhage and dismal outcome. PMID- 16358558 TI - Presynaptic effects of carbon monoxide in the myoneural synapse of the frog. PMID- 16358559 TI - The influence of a probiotic product on PC-12 cell differentiation. PMID- 16358560 TI - Specificity of internal lexicon in children with deviant behavior. PMID- 16358561 TI - Stevioside is a novel stimulator of pharmacon clathration with glycosides. PMID- 16358562 TI - Gender differences in verbal learning of 5- to 15-year-old children and adolescents. PMID- 16358563 TI - Purinergic P2X4 receptors are involved in the modulation of giant depolarizing potentials in the hippocampus of newborn rat. PMID- 16358564 TI - The biphasic effect of L-DOPA on the electric activity of an isolated dopaminergic neuron. PMID- 16358565 TI - Cadaverine-induced induction of superoxide dismutase gene expression in Mesembryanthemum crystallinum L. PMID- 16358567 TI - Peculiarities of phagocytosis in bottlenose dolphin (Tursiops truncatus) during the period of adaptation to the captivity conditions. PMID- 16358566 TI - Measurement errors in determining tropospheric bioaerosol concentrations in the southern region of western Siberia. PMID- 16358568 TI - Seasonal changes in the standing stock of essential polyunsaturated fatty acids in the biomass of phyto- and zoobenthos on a littoral station of the Yenisei River. PMID- 16358569 TI - Self-oscillations of water transport in the plant root. PMID- 16358570 TI - A novel structural type of plant cuticle. PMID- 16358571 TI - Genetic polymorphism of Siberian newt (Salamandrella keyserlingii, Caudata, Amphibia) in its range and the cryptic species of the newt S. schrenckii from Primorie. PMID- 16358572 TI - Pulse electric discharges and prolonged microbial resistance of water. PMID- 16358573 TI - Induction of lethal mutations of the leg-arista-wing complex gene of Drosophila melanogaster. PMID- 16358574 TI - Responses of the electrogeneration system of Synodontis (Mochokidae, Siluriformes) to weak electric fields. PMID- 16358575 TI - Cytogenetic study of the cornea epithelium of rodents inhabiting the areas of chemical contamination and radioactive pollution in the Oka-Don interfluve. PMID- 16358576 TI - A long-lived parasite extending the host life span: the pearl mussel Margaritifera margaritifera elongates host life by turns out the program of accelerated senescence in salmon Salmo salar. PMID- 16358577 TI - The insulating properties of the pelage of the North-American porcupine (Erethizon dorsatum): the influence of quill-like structures on heat transfer. PMID- 16358578 TI - Genetic divergence of the yeast Arthroascus fermentans in Japan. PMID- 16358579 TI - A mathematical model of the temperature conditions of mammalian burrows as exemplified by the burrow of the marmot Marmota bobak Mull., 1776. PMID- 16358581 TI - Estimation of population size stability in small mammals. PMID- 16358580 TI - Mechanism of defensive morph formation in Scenedesmus acutus (Chlorophycea, Scenedesmacea). PMID- 16358582 TI - Why does the yeast Kluyveromyces wickerhamii assimilates but not ferments lactose? PMID- 16358583 TI - Variation of distances between cilia is a novel mechanism of the regulation of mechanosensitivity. PMID- 16358584 TI - LOSK (SLK) protein kinase activity is necessary for microtubule organization in the interphase cell centrosome. PMID- 16358585 TI - Molecular factors required for human immunodeficiency virus type I infectivity. PMID- 16358586 TI - [The model of action in early schizophrenia]. AB - I will present the model of thinking of the Cracow society in dealing with first episodes of schizophrenia in adolescents. During adolescence schizophrenia brings about diagnostic difficulties, because it is a period of stormy psychosocial and biological changes in the patient. Also, the responsibility of the therapists seems to be different--perhaps even more so than in their work with an adult. Importance of the environment of the psychiatric ward and group psychotherapy as well as the peer group for the patients, the role of family therapy, especially trans-generational transmission, place of individual psychotherapy and biological therapy in this system, all will be discussed. I will describe the importance of both, pedagogical therapy and the school in the course of the therapeutic process of the adolescent patients. In a closer manner, I will also describe the emotional problems of therapists dealing with adolescents having psychosis. PMID- 16358587 TI - [Duration of untreated psychosis in first-episode schizophrenia: clinical and cognitive correlates]. AB - OBJECTIVE: Duration of untreated psychosis (DUP) is associated with poor premorbid functioning and poor outcome in patients with schizophrenia. The authors determined whether the duration of untreated psychosis was associated with severity of clinical symptoms and neurocognitive functioning in patients with first-episode schizophrenia. METHOD: A total of 30 first-episode patients were evaluated by means of WCST, Vocabulary, Arithmetic, Block Design, attention and verbal fluency tests. RESULTS: The mean duration of untreated psychosis was 31.3 weeks. The group was divided on the basis of median duration of untreated psychosis (10 weeks) to short and long DUP group. There were no significant differences between the groups in results of clinical symptoms and neurocognitive functioning. There were no correlations between severity of clinical symptoms and neurocognitive functioning with duration of untreated psychosis. CONCLUSIONS: In the group of patients with first-episode schizophrenia, the duration of untreated psychosis is not associated with severity of clinical symptoms and neurocognitive functioning. PMID- 16358588 TI - [Religiousness of patients with diagnosis of schizophrenia. Introductory studies]. AB - AIMS: (1) The correlations between various aspects of religiousness and age, span of treatment, rate of therapy, anxiety-state, anxiety-trait, locus of control, and self-rating of the state of self-feeling, and the state of feeling of illness, and quality of life in the group of patients with a diagnosis of schizophrenia and in the group of healthy persons, and (2) the comparison of various aspects of religiousness of patients with schizophrenia and healthy persons were studied. METHODS. The Individual Religiousness Scale, Scale of Christian Religiousness, Intrinsic, Extrinsic and Quest Religious Orientation Scales, Scale of Fundamentalism, Rotter's questionnaire of locus of control, State-Trait Anxiety Inventory, visual analogy of self-feeling, visual analogy of feeling of illness, questionnaire of quality of life--WHOQOL-BREF--were used in the study. 61 adults of both sexes, 30 patients with a diagnosis of schizophrenia according to ICD-10 and DCR-10 in the diagnostic group and 31 healthy persons were studied. Groups were similar in the terms of age, sex, education, abode, social conditions, professional status, civil state and denomination. RESULTS: It was noted, that: (1) intrinsic and extrinsic religiousness correlates with age in both groups and with span of treatment in the diagnostic group, but only intrinsic religiousness correlates with the rate of therapy; the locus of control increases with a decrease of orthodox religiousness in the diagnostic group and with ethical aspects of religion in the group of healthy individuals; state of anxiety is correlated backwards with orthodox religiousness in the diagnostic group; (2) patients diagnosed as having schizophrenia have a significantly higher level of extrinsic religiousness and fundamentalism than healthy individuals: (3) there was many results pointed out in bibliography, that had no verification in these studies. CONCLUSIONS: The knowledge about the religiousness of schizophrenia patients can be important for the diagnostic and therapeutic process in this disorder, therefore further studies on these characteristics are necessary. PMID- 16358589 TI - [Neurotrophic factors and their role in the pathogenesis of affective disorders]. AB - Neurotrophic factors are a group of proteins with a similar structure (The regulation of neuronal plasticity and neuron protection are some of their biological functions). The group of neurotrophic factors consists of: growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3) and neurotrophin 4/5 (NT-4/5). BDNF is the most important neurotrophin from the affective disorders point of view. Preclinical and clinical studies of altered BDNF expression during chronic stress and increased BDNF activity during antidepressant treatment, confirm the role of BDNF in the pathogenesis of depression. Studies on animal models point to the antidepressant effect of BDNF, similar to long-term antidepressant treatment. The intracellular mechanisms mediated by this neurotrophic factor are connected with signal transduction pathways in cells (mainly mitogen-activated protein kinase cascade and cyclic adenosine 3',5'-monophosphate cascade). The BDNF serum level studies suggest a correlation between the BDNF expression in the central nervous system and its serum levels, what could make BDNF levels specific markers of depression. The molecular genetic studies focus on associations between BDNF gene polymorphisms and bipolar disorder or cognitive functioning disturbances. The novel pathogenetic theories of depression based on neuronal plasticity (Duman et al.) and disturbances in neurogenesis (Kempermann and Kronenberg) can be a kind of recapitulation of research on the role of neurotrophins in depression. However many issues related to the role of neurotrophic factors in affective disorders are still unclear and determine areas of future scientific interests in this field. PMID- 16358590 TI - [The role of zinc in the pathogenesis and treatment of affective disorders]. AB - Zinc is an important modulator of glutaminergic transmission. Recent data indicate that pathology of amino-acidergic neurotransmission may contribute to mood disorders and may be involved in antidepressant-like actions in laboratory models. A growing body of evidence implicates a derangement of zinc homeostasis in mood disorders. The zinc deficiency is related to a variety of psychological symptoms especially depression. There are many reports indicating significant changes in blood levels of zinc during a depressive episode. Moreover zinc exhibits antidepressant-like and anxiolytic-like effects in animal models of depression, in rodents. Recent data revealed that zinc enhances the antidepressant effect in laboratory animals. This article reviews the alterations in central and peripheral zinc homeostasis in relation to pathophysiology and treatment of depression. PMID- 16358591 TI - [The role of copper and magnesium in the pathogenesis and treatment of affective disorders]. AB - Magnesium and copper are important modulators of NMDA-receptor activity. Recent data indicate that disturbances of glutamatergic transmission (especially via NMDA-receptor) are involved in pathogenesis of mood disorders. Magnesium deficiency, the same as disturbances in turn over of copper, are related to a variety of psychological symptoms especially depression. There are many reports indicating significant changes in blood levels of magnesium or copper during a depressive episode. Moreover magnesium exhibits antidepressant-like and anxiolytic-like effects in animal models of depression, in rodents. This article reviews the alterations in central and peripheral magnesium and copper homeostasis in relation to pathophysiology and treatment of depression. PMID- 16358592 TI - [Antidepressants and cytokines--clinical and experimental studies]. AB - Clinical and experimental studies indicate that stress and depression are associated with the up-regulation of the immune system, including increased production of pro-inflammatory cytokines. When administered to patients or laboratory animals, some of these cytokines induce typical symptoms of depression. It is known that cytokines modulate brain neurotransmission and the activity of the hypothalamic-pituitary-adrenal axis, both of which are disturbed in depression. This review summarizes in vivo, ex vivo and in vitro clinical and experimental studies of the effect of antidepressants on cytokine production. In vitro culture and animal studies in particular suggest that antidepressants of several classes decrease the production of pro-inflammatory cytokines and shift the pro/anti-inflammatory cytokine balance towards the latter. Some studies suggest that immunological disturbances, including changes in cytokine levels, are not shared by all depressive patients, which means that only in certain groups of patients may the immunomodulatory action of antidepressants play a significant role in producing the therapeutic effect. PMID- 16358593 TI - [Depression and pain: novel clinical, neurobiological and psychopharmacological data]. AB - Epidemiological studies confirm frequent appearance of pain symptoms in depressive patients and a marked prevalence of depression in pain conditions. These observations seem to point at a close intertwining between mood regulation and pain perception. In the pathogenesis of both depression and pain symptoms, an important role has been attributed to disturbances of serotonergic and noradrenergic neurotransmission as well as to neuropeptides such as opioids and substance P. In mood regulation as well as in the perception and emotional dimension of pain stimuli, such brain structures as the amygdala, anterior cingulate cortex and prefrontal cortex are of main significance. The action of antidepressant drugs results in a normalization of the activity of those neurotransmitter systems and brain structures. It was found that dual action antidepressants (i.e. influencing both serotonergic and noradrenergic system) such as tricyclic antidepressants and new generation drugs (venlafaxine, milnacipram, duloxetine, mirtazapine) exert a stronger antidepressant effect and possess a broader therapeutic spectrum, including also an effect on pain symptoms. These drugs have been also increasingly used for the treatment of pain symptoms in somatic illnesses (e.g. diabetic neuropathy, fibromyalgia). PMID- 16358594 TI - [Bipolarity among unipolar affective disorder patients--uniDEP-BI national multi site study]. AB - The main objective of the study was to evaluate the occurrence of bipolarity among outpatients with a recurrent major depressive disorder. METHOD: The uniDEP BI study was conducted throughout Poland by 96 psychiatrists from 16 sites, who were trained in the study instruments. The sample was selected from the population of outpatients with at least one depressive episode (n = 880). The final study group included 246 working age adults (75.2% of females) treated for recurrent unipolar disorder. The study questionnaire consisted in the DSM-IV criteria of major depressive episode, (hypo)manic episode, criteria for bipolar spectrum disorder by Ghaemi et al. RESULTS: Unipolar mood disorder was confirmed in 32.9% cases, bipolar I disorder was found in 19.5%, bipolar II in 35% and bipolar spectrum disorder in 12.6% of the assessed patients. Patients with bipolar compared to the unipolar mood disorder had significantly more frequently a family history of bipolar disorder, short (hypo)manic episodes after antidepressive treatment, premorbid cyclo-, hyperthymic or impulsive (borderline type) personality, recurrent depressive episodes, atypical depressive symptoms, early onset of depression (< 25 yrs), distracted attention and panic attacks. Subjects with confirmed recurrent depressive disorder were significantly more often working (37.2% vs. 22.7%). Duration of the illness was significantly shorter and the number of previous depressive episodes was significantly lower in this group. CONCLUSIONS: The findings showed that the bipolarity features are more common among patients with unipolar mood disorders. It also points to a need of proper and deeper diagnostics of affective disorders and verification of rules and period of antidepressive and normothymic treatment. PMID- 16358595 TI - [Depression in the course of bipolar disorder and recurrent depressive disorder in the elderly: diagnostic difficulties]. AB - Misdiagnosis of bipolar disorder in depressive patients is a common clinical problem estimated to be evident in up to 40% of patients. Elderly patients might be especially vulnerable to that sort of diagnostic error. AIM: To estimate the rate of misdiagnosis in the elderly (60yrs+) hospitalized due to depression and to establish clinical correlates that might improve diagnosis. METHOD: A retrospective analysis of medical records of all the patients hospitalized in the University based Psychogeriatric Ward and suffering from a depressive episode due to bipolar disorder or recurrent depressive disorder. RESULTS: The rate of misdiagnosis was 54% in bipolar and 9% in recurrent depressive disorder. Bipolar patients were mainly misdiagnosed as having recurrent depression. A severe episode was more common in bipolar subjects (particularly in women) while recurrent depressive subjects tend to suffer from less severe but more protracted episodes. Somatic symptoms of depression were more prevalent among recurrent depressive subjects while psychotic features, particularly delusions, and, to the lesser extent, hallucinations, were more common in patients suffering from bipolar depression. CONCLUSIONS: Bipolar depression is very often misdiagnosed in the hospitalized elderly. There are several features that might help the clinician to distinguish it from recurrent depression. Long history of illness, high number of previous episodes, severe episodes and the presence of psychotic symptoms are typical for bipolar elderly while a protracted current episode and the presence of somatic symptoms of depression might indicate the diagnosis of recurrent depression. PMID- 16358596 TI - [Vascular factors in dementia]. AB - Cerebrovascular factors are a common cause of dementia or contribute to cognitive decline in other dementias. Studies showing that cerebrovascular factors are the risk factors for neurodegenerative dementias, especially Alzheimer's disease. Practically all neurodegenerative dementias have a vascular component that reduces cerebral perfusion and has great impact on the clinical picture. Recent data support the view that the neurodegenerative process is caused by cerebrovascular mechanisms. The results showed that patients with vascular cognitive impairment have a typical clinical picture. Various important non cognitive features are caused by cerebrovascular factors and are associated with a more rapid course of illness. On the other hand the term vascular diseases or cerebrovascular factors include a variety of vascular pathologies. PMID- 16358597 TI - [Vascular factors and progression of cognitive decline in elderly people]. AB - The aim of the study was to assess the impact of vascular factors on the rate of progression of cognitive impairment. The study included 291 subjects without dementia. Cognitive function were assessed with the Alzheimer Disease Assessment Scale--cognitive subscale (ADAS--cog) which were conducted at baseline and at the end of the study. Statistical analysis included 215 persons. During the observation AD developed in 19 subjects and 11 vascular and mix dementia (according to DSM-IIIR and DSM-IV criteria). Subjects were categorized by the baseline Modified Hachinski Ischemic Score as having vascular factors 0-1 point (n = 140) or vascular factors > 1 point (n = 75). Statistical analyses were based on the patients' Modified Hachinski Ischemic Score dichotomization. The results show that vascular factors were risk factors for Alzheimer type dementia but the study does not prove the impact of vascular factors on progression of cognitive impairment. PMID- 16358598 TI - [Program for promoting social attachment among Poles in Australia in relation to their mental status]. AB - Among the Polish community in Sydney in 1989 there was a significantly higher percentage of men than women. Compared to the Australian population, there was also a high percentage of single people (especially men), a high rate of childlessness and a majority of incomplete families. The data on admission to psychiatric institutions in New South Wales indicate, that Polish-born persons (males) hospitalized with psychiatric disorders were 2.5 times more than the population of Australian-born. Sociologists pointed that because of lesser contacts with other members of their ethnic groups, people find themselves isolated. There were also limited family members or relatives who could provide support. From the South Western Sydney Area Health Service initiative a 2-year programme "Gazeta seniora". The strategic goal was to reduce number of isolated Polish-born older people through a number of regular initiatives: newspaper supplement, radio programmes, meeting groups, phone line. According to the information obtained from health and social services and from the respondents survey, letters and interviews, there was an increased access to medical services (15 percent), number of ethic meeting groups (40 percent). The programme has encouraged community leaders to provide personal contact and support for isolated people. After two years there were twice as many programme participants "satisfied with life" mainly because of "other peoples and the community's" involvement (from 7.0 to 21.4 percent). In the discussion part the authors stressed a holistic and long-term aspect of social and environmental support of any community based health promotion intervention related to psychic health improvement. PMID- 16358599 TI - Dose response of topical anesthetic on laryngeal neuromuscular electrical transmission. AB - OBJECTIVES: Our purpose was to determine the effect of a dose response to decreasing concentrations of topical anesthetic upon laryngeal neuromuscular electrical transmission. METHODS: We performed a prospective study at a neurolaryngology referral center. Forty-three patients were divided into 5 groups. Each patient underwent laryngeal electromyography (EMG) of a thyroarytenoid muscle before and 60 seconds after topical laryngotracheal lidocaine hydrochloride, normal saline solution, or nothing was applied. The pretreatment and posttreatment measurements were recorded with the same indwelling EMG electrode. Group 1 (n = 12) received 4% lidocaine, group 2 (n = 9) received 2% lidocaine, and group 3 (n = 8) received 1% lidocaine. Group 4 (n = 5) received topical normal saline solution instead of lidocaine. A fifth group (group 5, n = 9) had 2 EMG recordings measured, each separated by 60 seconds, without topical anesthetic. RESULTS: Groups 1, 2, and 3 showed significant decreases in the maximum peak-to-peak amplitude of the EMG recording (48.5%, 49.7%, 44.7%, respectively). Groups 4 and 5 failed to show a significant change in peak-to-peak amplitude after 60 seconds. There was no dose response change in EMG with decreasing lidocaine concentrations. CONCLUSIONS: All concentrations of lidocaine administered in this study decreased the laryngeal neuromuscular electrical transmission as measured by laryngeal EMG. This group of patients did not exhibit any dose response to anesthetic concentration. This finding is clinically significant for both diagnostic and therapeutic uses of laryngeal EMG preceded by administration of topical anesthetic. PMID- 16358600 TI - Prefabricated composite free flaps for tracheal reconstruction: a new technique. AB - OBJECTIVES: Successful laryngotracheal reconstruction requires both structurally supported tissue that withstands airway pressure changes and well-vascularized epithelial lining to prevent granulation and stricture formation. For circumferential defects, end-to-end anastomosis achieves favorable results, but for long-segment or large noncircumferential defects, no proven methods have emerged. Several animal studies describe prefabricated soft tissue flaps wrapped around synthetic materials or cartilage. However, prefabricated flaps have had very little use in human airway reconstruction. We present a patient with laryngeal stenosis and tracheostomy dependence following chemoradiotherapy for hypopharyngeal carcinoma. METHODS: In an attempt to widen the patient's laryngeal airway, a thyrotracheal autograft procedure, previously described by our institution, was performed. We transferred a segment of hemitrachea cephalad using the thyroid gland as a "vascular carrier," thus creating an 8-cm-long trough inferiorly that involved a 40% defect of the anterior tracheal circumference. Severe radiation damage to the cervical skin precluded use of traditional tracheoplasty methods. We used a technique whereby costal cartilage strips were implanted into a radial forearm free flap, designed to replicate the anterior tracheal wall. RESULTS: Four weeks later, we harvested the prefabricated composite flap and placed it into the defect, using forearm skin as tracheal lining. The cervical skin defect was closed with an island deltopectoral flap. A soft stent was kept in the neotrachea for 3 weeks, and a tracheostomy tube was left beneath it. The tracheostomy was subsequently closed with local advancement flaps, and the patient currently maintains an excellent airway. CONCLUSIONS: Prefabricated composite free flaps are an attractive option for certain challenging cases of airway reconstruction. PMID- 16358601 TI - Clinical characteristics of eosinophilic esophagitis in children. AB - OBJECTIVES: The role of eosinophilic esophagitis (EE) in aerodigestive tract disorders in children is underestimated and overlooked, primarily because of a lack of understanding of this disorder by otolaryngologists. We sought to better characterize the clinical presentation of EE in order to increase awareness among otolaryngologists. METHODS: We retrospectively reviewed 71 children with biopsy proven EE to determine the most common symptoms and laboratory findings that should increase the clinical suspicion of EE. RESULTS: Dysphagia, food impaction, and emesis were the most common symptoms in children with EE. Asthma was the most common airway diagnosis. Rhinosinusitis was the most common otolaryngological diagnosis. Food allergy was present in 60% of the children tested. Eighty-three percent of the children with elevated immunoglobulin E levels had thick linear streaking or patchy white exudate of the esophagus seen on esophagoscopy. Other major medical comorbidities existed in more than half of the children with EE, of which psychiatric disorders and other disorders of the aerodigestive tract were the most common. CONCLUSIONS: Eosinophilic esophagitis may contribute to treatment failure in patients with common and complicated aerodigestive tract disorders. To encourage clinicians to avoid overlooking the diagnosis, we present an evaluative algorithm to increase the suspicion of this entity. PMID- 16358602 TI - Cidofovir and the black box warning. PMID- 16358603 TI - Case of progressive dysplasia concomitant with intralesional cidofovir administration for recurrent respiratory papillomatosis. AB - Recurrent respiratory papillomatosis (RRP) is characterized by benign wartlike growths in the larynx with occasional spread to the lungs. A broad range of therapeutic measures have been used to treat RRP; the primary treatment is laser vaporization of the lesions. Recurrences of the disease are common, and alternate methods of treatment are being used to prevent recurrence, including cidofovir. Cidofovir is a cytosine nucleotide analog with antiviral properties that is approved by the US Food and Drug Administration for treatment of cytomegalovirus retinitis and is currently being used off-label for RRP. Cidofovir has gained initial success in slowing the rate of disease recurrence when used at the time of surgery. However, the use of cidofovir lends concern to several adverse side effects, including the potential for carcinogenesis. We report here a 28-year-old woman who was treated with intralesional cidofovir at the time of surgery over the span of 27 months. The initial pathology results demonstrated benign disease with progression to severe dysplasia during the treatment time. Cidofovir's potential for carcinogenicity remains largely undefined, and thus, we are currently undertaking a project involving the evaluation of sequential paraffin embedded samples of resections from a large cohort of patients with RRP treated at the University of Iowa Hospitals and Clinics. PMID- 16358604 TI - EYA1 expression in the developing inner ear. AB - OBJECTIVES: We sought to determine the developmental anatomy and EYA1 protein distribution in the inner ear of Xenopus laevis. METHODS: Xenopus laevis embryos were stained with monoclonal antibodies and imaged with confocal microscopy. RESULTS: At stage 27, the otocyst fully forms, with strong tubulin staining of early sensory cells at its ventromedial aspect. Neuronal ingrowth follows at stage 33/34. At stage 50, the semicircular canals are complete. EYA1 localizes to the anterior aspect of the otocyst from stages 37 to 44. By stage 50, EYA1 distribution is localized primarily to the sensory maculae and the endolymphatic duct of the developing inner ear. CONCLUSIONS: Whole mount confocal imaging of the developing Xenopus inner ear delineates the exact timing of otic development, sensory cell differentiation, and innervation. EYA1 protein expression has a distinct distribution pattern at the anterior aspect of the developing otocyst in stages 41 and 44. Later stages have a more localized pattern, in which EYA1 is detected only in the sensory epithelium and endolymphatic duct. This specific pattern of expression indicates a possible role in the determination of the anterior-posterior orientation of the inner ear, as well as a later role in sensory cell differentiation. PMID- 16358605 TI - Nonsyndromic isolated unilateral cochlear nerve aplasia without narrow internal auditorymeatus: a previously overlooked cause of unilateral profound deafness in childhood. AB - OBJECTIVES: Juvenile or adolescent unilateral profound sensorineural deafness (worldwide prevalence, 0.1% to 0.2%) has been attributed to postnatal viral infection, sudden deafness, prenatal and perinatal problems including maternal rubella and viral infections, congenital innerear anomalies, and other factors. Herein, 2 cases are reported and another potentially important cause of unilateral profound hearing loss is proposed. METHODS: Two nonsyndromic cases of a presently "very rare" cause of pediatric unilateral deafness are presented as a retrospective case study. RESULTS: The 2 patients showed isolated aplasia of the cochlear nerve; other branches of the eighth cranial nerve, the seventh nerve, and the inner ear were spared,and there was no anomaly of the internal auditory meatus. Both functional and imaging studies confirmed the isolated lesion (absence) of the cochlear nerve. CONCLUSIONS: Because of the absence of bony abnormalities, such cases may have been overlooked. The authors would like to advocate this isolated anomaly of thecochlear nerve as an important cause of juvenile or adolescent unilateral profound deafness. PMID- 16358606 TI - Radiology of the cochlear aqueduct. AB - OBJECTIVES: We sought to determine normative data for the radiologic presentation of the cochlear aqueduct (CA), hypothesizing that increasing the scanner's resolution could enhance detection capability. METHODS: Axial sections of 502 high-resolution computed tomography (CT) images of temporal bones (488 patients) were reviewed. A type 1 CA was visualized on CT scans up to the vestibule, and its portion in the otic capsule segment could be seen as a thin (<1 mm) streak. In type 2, we were able to detect the medial two thirds of the structure, but we failed to see the whole otic capsule portion. In type 3, only the external aperture of the aqueduct and/or the medial third was seen. We defined undetectable CAs as type 4. RESULTS: We obtained CT scans with 0.6-, 1.1-, or 1.3 mm-thick slices through the petrous bones in 9.5%, 58.8%, and 31.7% of cases, respectively. The CA was visible and bilaterally symmetric in 49% of the images, and type 2 was the most commonly detected CA type (36%). The CA was invisible on either side in 21.9% of scans, irrespective of CT resolution, and was asymmetric in 53 of the 502 images. The CA types varied with changes in resolution, although type 3 appeared unchanged independent of alterations in resolution in most cases. CONCLUSIONS: There was no significant difference in CT detection capability between CA types at different resolutions. Computed tomography failed to demonstrate any CAs > or =1 mm in width in the otic capsule segment. PMID- 16358607 TI - Interactive effects of high-pass filtering and masking noise on word recognition. AB - OBJECTIVES: This study sought to examine the word recognition performance in noise of individuals with a simulated low-frequency hearing loss. The goal was to understand how low-frequency hearing impairment affects performance on tasks that challenge temporal processing skills. METHODS: Twenty-two normal-hearing young adults participated. Monosyllabic words were presented in continuous and interrupted noise at 3 signal-to-noise ratios of -10, 0, and +10 dB. High-pass filtering of the stimuli at 3 different cutoff frequencies (ie, 1,000, 1,250, and 1,500 Hz) simulated the low-frequency hearing impairment. RESULTS: In general, performance decreased with increasing cutoff frequency, was higher for more favorable signal-to-noise ratios, and was superior in the interrupted condition relative to the continuous noise condition. One important revelation was that the magnitude of the performance superiority observed in the interrupted noise condition did not diminish with high-pass filtering; ie, the release from masking in interrupted noise was preserved. CONCLUSIONS: The results of the present study complement previous findings in which this paradigm was used with low-pass filtering to simulate a high-frequency hearing loss. That is to say, low frequency hearing channels are inherently poorer than high-frequency channels in temporal resolution. PMID- 16358609 TI - A recent proposal by the American Council on Graduate Medical Education. PMID- 16358608 TI - C-C chemokine ligand 2 gene expression in nasal polyp fibroblasts: possible implication in the pathogenesis of nasal polyposis. AB - OBJECTIVES: Recruitment of macrophages is essential to the pathogenesis of nasal polyps (NP), since this disease is inflammation-related. In this study, the effects of tumor necrosis factor alpha (TNF-alpha) on the expression of C-C chemokine ligand 2 (CCL2) in fibroblasts derived from nasal polyps (NPFs) were investigated. The roles of cyclooxygenase (COX) 2 and prostaglandins in the mediation of TNF-alpha-stimulated CCL2 gene expression were also investigated. METHODS: Northern blot analysis was used to study the expression of CCL2 and c Fos in cultured NPFs. An electrophoretic mobility shift assay was used to explore the interactions between activator protein 1 (AP- 1) and DNA. Immunohistochemistry was used to explore the in vivo expressions of COX-2, CCL2, and CD68 in NPs. RESULTS: The Northern blot analysis showed that TNF-alpha stimulated the expression of CCL2 and COX-2 genes, and the synthesis of CCL2 messenger RNA was COX-2-dependent. A transient elevation of c-Fos and c-Jun messenger RNAs was induced by TNF-alpha, whereas COX-2 inhibitors NS-398 and meloxicam abolished the up-regulation of c-Fos. The electrophoretic mobility shift assay revealed that TNF-alpha triggered AP-1 and DNA binding and again, NS 398 and meloxicam inhibited this reaction via reducing c-Fos synthesis. Curcumin (AP-1 inhibitor) markedly suppressed the TNF-alpha-induced CCL2 expression. The immunohistochemical staining of NP surgical specimens also revealed an intimate alignment between CCL2-positive fibroblasts and CD-68-positive macrophages. CONCLUSIONS: These data suggest that NPFs may contribute to NP development by synthesizing CCL2 to promote macrophage recruitment. Furthermore, COX-2 facilitates CCL2 transcription in NPFs via a c-Fos and AP-1 signaling pathway. PMID- 16358610 TI - Complications of mastoiditis in children at the onset of a new millennium. PMID- 16358611 TI - [Two post-menopausal women with vaginal bleeding due to non-gynaecological malignancies]. AB - A gynaecological examination was performed because of vaginal bleeding in a 67 year-old woman who was admitted for dyspnoea and a painful cough. A tumour was detected in the vaginal wall that turned out to be a metastasis of a locally extensive and metastasised Grawitz tumour. In a 76-year-old woman who presented with vaginal bleeding, speculum examination revealed a polyp that protruded from the cervix. A non-Hodgkin lymphoma was demonstrated histopathologically. In both patients, the cervical smear showed Pap-I. Both patients were given palliative treatment. According to the guidelines, the policy of general practitioners and gynaecologists in cases of postmenopausal blood loss should be directed primarily at the exclusion of malignancy by means of transvaginal echoscopy. However, some abnormalities may then be missed. A complete gynaecological examination then becomes very important. Speculum examination forms an essential part of such a work-up. PMID- 16358612 TI - [The practice guideline 'Heart failure' (first revision) from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - The practice guideline 'Heart failure' from the Dutch College of General Practitioners will be a source of support for the general practitioner/family physician. Its clear set of diagnostic steps helps the general practitioner to assess heart failure in patients with breathlessness, fatigue, decreased exercise tolerance or ankle swelling. Testing plasma concentrations of natriuretic peptides plays a new and central diagnostic role. This guideline differs from the previous one in another important aspect. Based on many RCT's and reviews the authors have succeeded in developing a detailed but balanced treatment protocol for heart-failure patients in general practice. However, adverse effects from co- and multi-medication in the growing number of frail elderly may reduce the patients' compliance. PMID- 16358613 TI - [The practice guideline 'Heart failure' (first revision) from the Dutch College of General Practitioners; a response from the perspective of cardiology]. AB - The Dutch College of General Practitioners' practice guideline 'Heart failure' provides a clear insight into the history, diagnosis and treatment of patients with chronic heart failure. The revised guideline does however warrant some minor comments. It suggests that an elevated BNP value contributes towards the diagnosis of heart failure, however taking into account the high negative predictive value, BNP should, for the time being at least, only act as an 'instrument of exclusion' in the diagnosis of heart failure. The section on pharmacological treatment could have been expanded with the additional information that ACE-inhibitors should be replaced by angiotensin-II receptor blockers ifangioneurotic oedema appears. Another recommendation might be that early treatment with ACE-inhibitors and statins in high-risk patients reduces the incidence of heart failure. Furthermore, the guideline does not provide information about non-pharmacological treatment such as biventricular pacing and implantable cardioverter defibrillators. Prevention is not dealt with even though it is a task particularly suited to the GP. Lastly, it could have been more clearly pointed out that outpatient clinics providing special care for heart failure patients are well-recognized support facilities for patients with heart failure. In conclusion, the first revision can be considered as an adequate and workable practice guideline for the GP. PMID- 16358614 TI - [Hiccups]. AB - Hiccups are a common phenomenon but little is known about their pathophysiological mechanism or function. The concept of a hiccup reflex lacks a scientific basis. Closure of the glottis may be caused by an active contraction or by a passive, sudden myoclonic contraction of the inspiratory musculature, leading to a rapid reduction in intrathoracic pressure. Hiccups are often benign and of short duration but they can sometimes be a serious medical problem due to their chronicity and underlying causes. Many remedies for hiccups have been reported, such as manoeuvres, medical treatment and invasive procedures. Chlorpromazine is the only drug that is registered for the treatment of persistent hiccups but it has many side effects. Baclofen is often used in practice, and favourable experiences with piracetam are known. PMID- 16358615 TI - [Preventive surgical prcedures for inherited risk of breast cancer]. AB - Preventive surgical procedures for inherited risk of breast cancer Forwomen with a demonstrated BRCA1 or BRCA2 mutation, the cumulative risk of developing invasive breast cancer before the age of 70 years is about 50-85% and the risk of developing invasive epithelial ovarian cancer is 20-60%. Regular surveillance including physical examination and imaging is offered to mutation carriers and the options for risk-reducing surgery are discussed. Although bilateral prophylactic mastectomy is a drastic intervention, it significantly reduces the incidence of breast cancer. For mutation carriers with breast cancer, the decision to combine risk-reducing surgery with treatment is determined by the TNM stage of the disease. Prophylactic bi- or contralateral mastectomy after previous treatment for unilateral breast cancer reduces the incidence of contralateral breast cancer, but has no impact on survival. The complexity of the problem demands a multidisciplinary approach within the context of a family cancer clinic. PMID- 16358616 TI - [Summary of the practice guideline 'Heart failure' (first revision) from the Dutch College of General Practitioners]. AB - The guideline entitled 'Heart failure' from the Dutch College of General Practitioners has been revised. The general practitioner makes the diagnosis of heart failure in a patient with the core symptoms (dyspnoea, fatigue, oedema) in combination with paroxysmal nocturnal dyspnoea, orthopnoea, crepitations, elevated central-venous pressure, a third heart sound or ifictus cordis is visible outside the mid-clavicular line. Further investigations include a panel of laboratory investigations, an ECG and possibly echocardiography and radiographic chest investigations. One new recommendation is that on suspicion of heart failure the plasma concentrations of B-type natriuretic peptide (BNP) or N terminal pro-BNP should be investigated. The step-by-step medication plan has been changed; beta-blockers have been introduced to the plan. In the treatment of acute heart failure, rapidly working sublingual nitrates should be given first and foremost. PMID- 16358617 TI - [From the Cochrane Library: brushing the teeth with an electric toothbrush with an oscllating rotating movement more effective against plaque and gingivitis than brushing with a conventional toothbrush]. AB - Removing dental plaque plays a key role in maintaining oral health and reducing gingivitis and parodontitis. A systematic Cochrane review ofrandomised controlled trials encompassing participants from the general public with uncompromised manual dexterity showed that when compared to the use of manual toothbrushes, the use of powered toothbrushes with a rotating oscillation action for a period of three months reduced gingivitis and (although not statistically significantly) plaque. The long-term clinical relevance of the differences is unclear. Electric toothbrushes that did not have an oscillating rotating movement were not consistently better than manual toothbrushes. PMID- 16358618 TI - [Gynaecological diagnosis of postmenopausal women with abnormal vaginal bleeding: a comparison with the guideline]. AB - OBJECTIVE: The Dutch guideline for obstetrics and gynaecology recommends transvaginal echography for all patients with postmenopausal vaginal bleeding, followed by histological examination in case the double endometrium layer is either > 4 mm or cannot be measured. We assessed whether this guideline was applied in clinical practice. DESIGN: Prospective. METHOD: Data were collected on the diagnosis of 837 patients with post-menopausal vaginal bleeding who were seen in the gynaecology clinics of eight hospitals during the period 1 January 2001-31 December 2002. Subsequently, the actual practice was compared with the guideline, which was known in all the hospitals at the start of the study. RESULTS: Transvaginal echography was performed in 818 of 837 (98%) women. The thickness of the endometrium could not be measured in 74 (9%) patients, was < or = 4 mm in 361 (43%) women and > 4 mm in 383 (46%) women. In 56 (7%) patients, including 3 who refused further diagnostics, a histological diagnosis was not obtained even though it should have been according to the guideline. A histological diagnosis was obtained in 118 (13%) patients in whom this was not necessary according to the guideline. In 86 (10%) patients, a hysteroscopy and/or curettage was performed, even though a histological diagnosis by endometrial aspiration might have sufficed. CONCLUSION: Adherence to the guideline on the diagnostic management of postmenopausal bleeding was fairly good. Overdiagnosis was seen more frequently than underdiagnosis. The efficiency of the diagnostic management in women with postmenopausal bleeding might be increased by limiting histological examination to women with an increased endometrial thickness, and by relying on endometrium aspiration in these women. PMID- 16358619 TI - [Abscess formation due to lost stones during laparoscopic cholecystectomy]. AB - Two women aged 79 and 69 years presented with abdominal pain at 15 and 38 months respectively after a laparoscopic cholecystectomy. In both cases perforation of the gallbladder had occurred with spillage of bile and gallstones. A CT-scan carried out at presentation showed signs of an abdominal abscess. The origin of the abscess was initially unknown. The first patient was operated on immediately. In the second patient a drain was placed in the abscess under ultrasound guidance. During operation gallstones were seen in the abscess cavity in both patients. Both patients recovered well. Although perforation of the gallbladder with spillage of gallstones during laparoscopic cholecystectomy often occurs, the incidence of major postoperative complications is low. However, in some cases reoperation is necessary. Removal of all spilled stones is therefore indicated to prevent complications. If abdominal symptoms persist after laparoscopic cholecystectomy, an abscess due to spilled gallstones should be considered. Ultra sound or CT are the most sensitive means of for tracing spilled gallstones or abscesses. PMID- 16358620 TI - [Severe psychosis in an African woman due to the antiretroviral agent efavirenz]. AB - A 34-year-old woman originally from Ghana was given efavirenz as antiretroviral therapy. One week later she was found to be in a psychotic state with paranoid hallucinations and anxiety; she then stabbed a nurse. The literature indicates that female patients of African origin appear to be more susceptible to the side effects of efavirenz due to genetically reduced clearance and therefore higher serum levels. PMID- 16358621 TI - [A careful course of action in a conflict regarding useful treatment of a newborn infant with severe brain damage]. AB - In a newborn female infant, it was concluded that severe perinatal asphyxia had caused such extensive cerebral damage that further medical treatment was useless. Based on their religious beliefs, the parents disagreed, despite the fact that the requested second opinions supported the conclusion of the medical staff. Since the parents persisted, a period of inurement was agreed upon during which reanimation would be performed if necessary. After several months, there was no change in the attitude of the parents towards the policy not to reanimate, even though it was clear that there was no improvement whatsoever in the patient's neurological status, while everyone agreed that she showed signs of increased suffering. The decision regarding the determination of a situation in which further medical treatment was useless was re-evaluated carefully. In a legal procedure started by the parents, the judge supported the decision of the attending physicians. In order to prevent the parents from taking their child home, in which case a situation could arise in which she would be deprived of adequate sedation or analgesia, which the attending physicians were obliged to provide, the Dutch Child Protection Council was consulted and the parents were deprived of their parental authority. Ultimately, the patient died suddenly due to respiratory and circulatory arrest without another situation in which reanimation might have been indicated. PMID- 16358622 TI - [Lithium, a potentially dangerous drug]. PMID- 16358623 TI - [Medical end-of-life decision-making for neonates and infants in the intensive care unit in the Netherlands]. PMID- 16358624 TI - [Summary of the practice guideline 'The STD consultation' from the Dutch College of General Practitioners]. PMID- 16358625 TI - [Summary of the practice guideline 'The STD consultation' from the Dutch College of General Practitioners]. PMID- 16358626 TI - [Count down for the influenza pandemic runs. Virus alarm: are we equipped?]. PMID- 16358628 TI - [Incidental findings in epigastric sonography. What is clinically relevant?]. PMID- 16358627 TI - [Influenza prevention in your practice (interview by Waldtraud Paukstadt)]. PMID- 16358629 TI - [Healing with bacteria? Studies prove the positive effects of probiotica in gastrointestinal diseases]. PMID- 16358630 TI - [Cardiovascular risk factors treated too infrequently: are family practitioners too negligent? (interview by Judith Neumaier)]. PMID- 16358631 TI - [Diagnosis and treatment of hypertension. Individual aims, flexible control]. PMID- 16358632 TI - [Hypertension and renal failure--a deadly combination]. AB - Kidney insufficiency is considered to represent a highly significant risk factor for cardiovascular disease, even when the glomerular filtration rate is only mildly reduced. Renal function has an influence on the prognosis that is independent of such conventional risk factors as hypertension and lipid metabolic disorders. Hypertensive patients suffering from renal insufficiency need particularly careful management to lower the cardiovascular risk in this special group of patients. Blood pressure targets in hypertensives with renal insufficiency are required to be lower than in patients with essential hypertension; current evidence points to a target of 125/75 mmHg. PMID- 16358633 TI - [Hypertension and stroke: new publications in 2005]. AB - Current studies confirm the previous European recommendations according to which the individual evaluation of the patient's risk is superior to a breakdown into general therapeutic groups. Studies on primary prevention show that patients with atrial fibrillation in particular obtain greater benefit from losartan than from atenolol, and that even a small reduction in blood pressure reduced the stroke risk. The greatest benefit was seen in those patients in whom the target pressure was achieved within a period of six months. The combination of a calcium antagonist and an ACE inhibitor prevented significantly more strokes than the combination of a beta blocker and a diuretic. In studies investigators secondary prevention a sartan was found to prevent more cardiovascular events than nitrendipine. PMID- 16358634 TI - [New tendencies in combination antihypertensive therapy]. AB - In the management of hypertension, the constant lowering of target pressures ever more frequently requires combination treatment in order to achieve the goals set. A combination of amlodipine and perindopril effectively lowered the blood pressure, and significantly reduced overall mortality, cardiovascular mortality, cardiovascular events and stroke risk in comparison with atenolol/bendroflumethiazide. In combination with both ACE inhibitors and AT1 blockers, diuretics reduced blood pressure more effectively that the respective components used alone. Now, however, the long-term use of diuretics is suspected of increasing the risk of terminal kidney failure and diabetes. This also means an enhanced probability of a cardiovascular event. In the ASCOT study, the combination of beta blocker and diuretic proved inferior to treatment with a calcium antagonist and ACE inhibitor. For the long-term application of diuretics as antihypertensive monotherapy, therefore, further comparative studies involving the latest antihypertensives are mandatory. PMID- 16358635 TI - [Management of malignant melanoma. Careful removal, consistent after-care]. PMID- 16358636 TI - [Cave: bacterial superinfections in neurodermatitis. On the skin of atopic patients streptococci are especially content]. PMID- 16358637 TI - [Bulimia nervosa--a modern eating disorder]. AB - Bulimia nervosa is characterized by episodes of compulsive eating of large amounts of food that are followed by measures undertaken to avoid weight gain, and an exaggerated concern about one's figure and weight. Bulimic symptomatology comprises a complex of various interacting components such as poor self-esteem, a negative attitude towards one's own body, eating behavior and current conflicts. Treatment must take account of all of these factors, and involves both internistic and psychotherapeutic elements. The severity of the condition, possible comorbidities, and psychosocial criteria are crucial for deciding whether treatment should be on an ambulatory or an inpatient basis. PMID- 16358638 TI - [Analog insulin preparations in patients with type 2 diabetes. Better rapid and brief than sluggish and long]. PMID- 16358639 TI - [Individualized therapy. Insulin therapy requirements in pregnancy and old age]. PMID- 16358640 TI - [More income, fewer expenses. Family physician and economist Dr. Riedl demonstrates how he does it (interview by Anke Thomas)]. PMID- 16358641 TI - [A flood of new contracts--criticism of personal association. Colleagues are fed up with paperwork]. PMID- 16358642 TI - [Closed practices, demonstrating colleagues. Why physicians envy bakers]. PMID- 16358643 TI - [Effective pain therapy with good tolerability. New opioid administration form by transdermal patch]. PMID- 16358644 TI - [Fatigue and dry cough. Sarcoidosis]. PMID- 16358645 TI - [Good leech--good outcome]. PMID- 16358646 TI - [Nobel [corrected] prize for the discoverers of Helicobacter pylori--the overlooked bacteria]. AB - Robin Warren and Barry Marshal won the Noble prize of 2005 in medicine for their discovery of the bacterium Helicobacter pylori and its role in gastritis and peptic ulcer disease. The historical review in the article shows that the bacteria was observed in the past, but wasn't accepted by the scientific community. The article, with regard to Kuhn's model for scientific revulotions, demonstrates how the prevailing dogmas about stress as the cause of ulcers and the sterility of the stomach, led researchers to overlook the bacterium for many years, despite evidence to its presence. The outlook shows the axiom that a paradigm shift that follows a scientific discovery is based upon an infrastructure of many factors rather than on pure rational science. This review inhances the historical lesson that facts and evidence are culturally dependent, and that critical physicians and researchers are essential for the progress of medicine and science. PMID- 16358647 TI - [Intravitreal triamcinolone acetonide for diffuse diabetic macular edema--one year follow-up]. AB - BACKGROUND: Macular edema is the main cause of visual impairment in diabetic patients. Its treatment is mainly based on laser photocoagulation. Intravitreal triamcinolone acetonide (TA) has recently been proposed as a new treatment for eyes with diabetic macular edema resistant to conventional laser photocoagulation. AIM: To evaluate the one year efficacy and safety of a single TA injection administered for diffuse diabetic macular edema unresponsive to prior laser treatment. DESIGN: Interventional case series. METHODS: Setup: University medical center out-patient clinic. PARTICIPANTS: Twenty-one patients with bilateral diffuse diabetic macular edema. INTERVENTION: A single intravitreal injection of triamcinolone acetonide. MAIN OUTCOME MEASURES: Visual acuity measured by ETDRS score, retinal area evaluation clinically and retinal thickness evaluation by Retinal Thickness Analysis (RTA) at 3 and 12 months following injection. Secondary outcomes were intraocular pressure control and cataract progression. RESULTS: Three months following injection, the mean improvement in visual acuity was 4.7 +/- 11.7 letters in the study group as compared to 0.2 +/- 11.4 in the control group (p = 0.18). No difference was noticed one year following injection. Clinical assessment of the retinal area of edema revealed a substantial difference between patients and controls at 3 months (p = 0.0006) and at one year (p = 0.05). RTA evaluation revealed improvement in retinal thickness solely at the 3 months exam. Four eyes developed high intraocular pressure and required treatment (p = 0.054). No difference in cataract progression was noted between the two groups (p = 0.69). CONCLUSIONS: Injection of TA may improve visual acuity for a limited time. Increased intraocular pressure is a frequent side effect. During a one year follow-up, no evidence of ocular toxicity was noted in eyes with diffuse diabetic macular edema. PMID- 16358648 TI - [Diabetic retinopathy following cataract surgery]. AB - BACKGROUND: Approximately 20% of patients undergoing cataract surgery have diabetes mellitus. AIM: To evaluate the course of diabetic retinopathy after cataract surgery. METHODS: Diabetic patients with no or mild to moderate preoperative diabetic retinopathy were included and classified into 4 groups (A D): A--The course of retinopathy, B--macular edema, C--the effect of voltaren ophtha eye drops and D--systemic glycemic control. Group E included eyes with previous laser treatment for proliferative retinopathy. Clinical and angiographic retinal findings were scored before and after surgery. Progression was defined as an increase in the retinal score. In groups A, C, D and E the non-operated eye served as a control. In group B, the eyes of nondiabetic patients who had undergone cataract surgery served as a control. RESULTS: Retinopathy was stable in 66% and progressed in 34% (p < 0.005). Progression occurred during the first 6 postoperative months in 84%. Preoperative retinopathy was a risk factor for progression. Good visual acuity was achieved in 67% and was correlated with: preoperative retinopathy and postoperative deterioration. Macular edema was found in 50% of eyes compared to 8% of the controls (p < 0.005). Its development was correlated with preoperative retinal status. Twenty six eyes were treated with voltaren eye drops and 24 with placebo. Progression of macular edema was seen less often in eyes treated with voltaren (p < 0.001). Deterioration of retinopathy was less common in cases when HbA1C was equal to or lower than 7.5 mg%. CONCLUSIONS: Close retinal follow-up after cataract surgery is recommended, especially in patients with preoperative diabetic retinopathy. Systemic control of diabetes and antiinflammatory eye drops may improve surgical results. PMID- 16358649 TI - [Cataract surgery and incidence of postoperative endophthalmitis in Israel]. AB - BACKGROUND: Cataract surgery is the most common surgery in ophthalmology. Acute postoperative endophthalmitis is one of the most serious complications of cataract surgery and may result in severe visual impairment and even loss of the eye. In Israel there is no established and controlled registry on the number of operations or the rate of postoperative complications. AIM: To survey the total number of cataract operations performed annually in Israel, to investigate trends in surgical techniques and intraocular lenses used, and to study the rate of postoperative acute endophthalmitis. METHODS: Since the year 1990, a questionnaire regarding cataract surgery has been sent to all of the recognized inpatient and ambulatory surgical centers in the country. As of the year 2000, a questionnaire regarding postoperative endophthalmitis was added. RESULTS: The number of cataract operations performed annually demonstrates a constant increase in the last decade. In the last survey, conducted in 2004, there was a total estimated number of 39,000 operations. The incidence of post cataract endophthalmitis ranged between 0.13%-0.25%, with a downwards tendency in the past years. CONCLUSIONS: The constant increase in the number of cataract operations probably reflects both a true increase in the rate of operations, as well as better retrieval of data. The number of surgeries relative to the size of the population of Israel is within the lower limits of the range accepted in the industrialized world. The incidence of acute postoperative endophthalmitis is similar to that reported in controlled series. There is a need for an established controlled registry including information on the number of operations and the rate of complications. PMID- 16358650 TI - [Ocular manifestations of rickettsial diseases]. AB - Rickettsial infection may cause severe systemic disease. The ocular manifestations of rickettsial infection may include sub-conjunctival hemorrhages, corneal abscesses, uveitis, optic nerve head edema, retinal vasculitis hemorrhages and infiltrates and endophthalmitis. Ocular complications may result in significant visual loss. Ocular manifestations may precede other systemic signs and symptoms and assist in clinical diagnosis. This report describes two cases with ocular involvement of rickettsial disease. PMID- 16358651 TI - [Upper lid frontalis suspension as a treatment for essential blepharospasm]. AB - BACKGROUND: Essential blepharospasm is a progressive disease of unknown etiology causing a significant decline in the quality of life of patients suffering from the disease. Currently used treatment for this condition is repeated injections of Botulinum toxin. Some of the patients' therapeutic response to Botox injections is insufficient, and some develop ocular complications. GOALS AND METHODS: This report presents our experience with sling frontalis suspension for essential blepharospasm in 4 patients with follow-up periods of six months to 2 years. A literature review is presented and discussed. RESULTS: A treatment trial with Botox injections was attempted in all four patients. Three of the patients showed insufficient treatment response. Three of the patients developed side effects of recurrent episodes of upper lid ptosis following injections. In all four patients that underwent operations significant reduction of blepharospasm was observed. In two of the patients Botulinum toxin could be discontinued, in one patient treatment intensity could be reduced, and in three--treatment complications were prevented. All four patients reported a high degree of satisfaction from treatment results. CONCLUSIONS: Our results indicate that sling frontalis suspension for essential blepharospasm is a safe and efficient treatment for essential blephrospasm with a high degree of patient satisfaction. Further large-scale studies are needed to substantiate our results. PMID- 16358652 TI - [Oxygen effect on ocular lens]. AB - BACKGROUND: Cataract is the leading cause of preventable blindness worldwide. Clinical observations and laboratory results have shown that oxygen has a possible toxic role in cataract formation. AIM: The aim of the present study was to demonstrate, measure and characterize the damage caused to bovine lenses in organ culture as a result of their exposure to hyperbaric oxygen pressure. MATERIALS AND METHODS: Twenty bovine lenses exposed to hyperbaric pressure were compared to 20 control lenses. Lenses were kept in an organ culture for 14 days. Each day the focusing ability of the exposed lenses was compared to controls. The comparison was based on the amount to which the focus point of each measured ray diverged from the focus point of the lens. Lenses were also examined under the microscope and morphologic changes in study lenses were compared to controls. RESULTS: A statistically significant difference in focusing ability between the study and control lenses was observed. The difference became larger during the incubation period indicating an accumulation of damage. The damage resulted from the peripheral but not the central part of the lenses. The morphologic changes observed under the microscope matched the damage profile of the focusing ability. CONCLUSIONS: Oxygen has a possible role in cataract formation. The effect of oxygen is cumulative. The route of damage formation follows the diffusion of oxygen into the lens. PMID- 16358653 TI - [Outcome of combined penetrating keratoplasty with vitreoretinal surgery using temporary Eckardt keratoprosthesis]. AB - BACKGROUND: For the adequate management of vitreoretinal pathology, good visualization through a clear cornea is needed to permit proper surgical repair. However, an opaque cornea, precludes this necessary visualization and makes posterior segment surgery impossible. Temporary keratoprosthesis facilitates visualization of intraocular structures in eyes that might otherwise be inoperable due to corneal opacities. AIM: To report the effectiveness of a combined procedure (pars plana vitrectomy with temporary Eckardt keratoprosthesis, vitreoretinal surgery, and penetrating keratoplasty) and to assess the functional outcome of eyes undergoing these procedures. METHODS: Six eyes from six consecutive patients operated on between 2001 and 2004 at the Department of Ophthalmology, Soroka University Medical Center, Beer Sheva, and followed-up for at least 6 months, were evaluated retrospectively. The evaluation focused on ocular history, visual acuity (VA), anatomical outcome and complications. RESULTS: The average follow-up period was 26 months. Prior to the operation, the most frequent diagnosis in the anterior segment was corneal scar in 4 eyes (67%). In the posterior segment, the most frequent diagnosis was retinal detachment in 5 eyes (83%). Preoperative VA was full light perception or less in all eyes. In five cases (83%), final postoperative VA was 6/240 or better. The corneal graft remained clear in five cases (83%). The retina remained attached in all cases. CONCLUSIONS: Combined corneal and retinal procedures using temporary keratoprosthesis are a safe and efficacious way of managing complicated corneal and retinal disease. These methods can preserve ambulatory vision and improve the quality of life of patients. Visual prognosis is usually limited due to the primary ocular disease. PMID- 16358654 TI - [Optic disc drusen and acute vision loss]. AB - Optic disc drusen (ODD) are calcified nodules within the optic nerve head. These are products of degenerated retinal ganglion cells axoplasmic transport, also known as hyaline or colloid bodies. They are mainly encountered as an incidental benign finding, or as a benign cause of swollen discs (in the differential diagnosis of papilledema). The majority of the patients are asymptomatic, and a slowly progressive sub-clinical visual field loss is found in about 80% of the patients. However, acute painless symptomatic visual field loss, occasionally massive, is rare. This is a case history of 4 patients who developed acute painless visual field loss which was attributed to optic disc drusen. The clinical features and proposed ischemic pathophysiology of acute vision loss and ODD are discussed. PMID- 16358655 TI - [Selective laser trabeculoplasty treatment for medication-refractory open angle glaucoma]. AB - BACKGROUND: Selective laser trabeculoplasty (SLT) is a new method to reduce intraocular pressure in eyes with primary open angle glaucoma. The laser parameters are set to selectively target pigmented trabecular meshwork (TM) cells without damage to the adjacent non-pigmented tissue. PURPOSE: A clinical retrospective study was conducted to evaluate the 12 months results of SLT in the treatment of medically uncontrolled open angle glaucoma. PATIENTS AND METHODS: During the period March to September 2004, the authors performed a SLT in 50 patients (50 eyes) with open angle glaucoma uncontrolled on maximally tolerated medical therapy. Treatment was carried out with a frequency-doubled, Q-switched Nd:YAG laser 532 nm.(Selectra 7000 Laser Coherent, Inc., Palo Alto,CA). Approximately 85 to 90 non-overlapping laser spots were placed over 180 degrees of the trabecular meshwork at energy levels ranging from 0.6 to 1.4 mJ per pulse. In patients who required additional SLT therapy, the untreated 180 degrees was treated. During the follow-up period, patients were treated with antiglaucoma medications as required. The success rates were defined as decreases in intraocular pressure (IOP) of 3 mmHg or more with no additional medications, laser, or glaucoma surgery. RESULTS: The mean IOP reduction from baseline 6 months after treatment was 21% and 20% after 12 months. The success rates were 66% after 6 months and 55% after 12 months. Four eyes (8%) did not respond to SLT. One hour after SLT, an increase in IOP of more than 5 mm Hg was detected in 5 eyes (10%). Seven patients (14%) required additional SLT. CONCLUSIONS: SLT is efficient in lowering IOP in medication-refractory open angle glaucoma. It should be considered in such patients when surgery is contraindicated or refused. PMID- 16358656 TI - [Lamellar macular hole and continuous vitreofoveal adherence]. AB - PURPOSE: To demonstrate a sub-type of idiopathic inner lamellar macular hole (ILH) that may merit surgical consideration. METHODS: The study was conducted on patients examined with optical coherence tomography (OCT) between August 2001 and December 2003 and included 22 patients each with ILH in one eye. Eyes with persistent vitreofoveal adherence at the ILH site were included in the study. Eyes that had previously undergone posterior vitrectomy and removal of the posterior hyaloid were excluded. RESULTS: Out of the 22 cases, persistent vitreous adherence to the edge of the ILH was detected in 3 eyes (3 patients; 13.6%). The ILH occurred spontaneously in 2 eyes and was detected after extraction of an intraocular foreign body in one eye. The vitreous adherence at the ILH, related to the incompletely detached posterior hyaloid in each, was asymmetric: in 2 eyes it was detached only nasal to the ILHs; in the third eye the posterior hyaloid was detached for 360 degrees around the adherence site, but appeared much thicker at a specific site than elsewhere. CONCLUSIONS: The findings indicate that persistent vitreofoveal adherence might be present in eyes with ILH. Further studies should signify whether such ILHs have a potential of progression, cases in which surgical intervention should be considered. PMID- 16358657 TI - [Indications--intravenous immunoglobulins--IVIG]. PMID- 16358658 TI - [Obesity is a risk factor for eye diseases]. AB - Obesity is the most prevalent metabolic disease in developed countries and its prevalence rates worldwide are increasing rapidly. The disease is associated with a considerably enhanced morbidity in many body systems as well as mortality. This article reviews the effects of obesity on the four major blinding eye diseases: age-related macular degeneration, diabetic retinopathy, cataract and glaucoma, and shows that obesity is a significant risk factor for these diseases and hence to blindness. PMID- 16358660 TI - [Image of the month. Splenic cyst]. PMID- 16358659 TI - [The treatment of mood stabilizers in children and adolescents suffering from bipolar affective disorder]. AB - Bipolar disorder is defined as a mood disorder. It is characterized by alteration in mood, from elation and/or irritability to depression. The prevalence of this disorder in children and adolescents is 1%, and it disrupts the lives of children and adolescents. The treatment of bipolar disorder includes mood stabilizers. In contrast to the extensive literature in adult bipolar disorder, controlled studies of lithium and anticonvulsants in the management of mood disorders in childhood are scarce. This review summarizes recent clinical pharmacologic studies of mood stabilizers, including lithium and anticonvulsants in the management of bipolar disorder in children and adolescents who suffer from this syndrome. In addition, the authors review new anticonvulsants such as lamotrigine, gabapentin and topiramate as mood stabilizers. PMID- 16358661 TI - [How I treat. . . evanescent youth. Dating back using chemical peels]. AB - Chemical peels induce the destruction and exfoliation of the epidermis using selected caustic substances. The specific properties of these agents result in a limited and controlled destruction of epidermal layers and of the superficial dermis. The choice of the appropriate method, the preparatory phase and the post peeling care are very important to reach optimal results. Some types of peeling are routinely used. We briefly report those designed for the superficial and medium depth peelings. The topical application of glycolic acid at high concentration induces a superficial peel resulting in shading of small wrinkles. Trichloracetic acid used at an appropriate concentration acts more deeply and thus represents a good indication for more severe signs of ageing. These peels have been used for decades by dermatologists to improve the visible signs of skin ageing and to treat some alterations of the cutaneous relief. PMID- 16358662 TI - [Rounded atelectasis: a follow up of 10 years by CT]. AB - We report the case of a 69-year-old man who presented with a rounded atelectasis (RA). During a 10-year follow-up by Computed Tomography (CT), the lobe showed a progressive shrinkage, and a moderate increase in size of the lesion led to a percutaneous biopsy which confirmed the diagnosis. RA is an unusual form of lung consolidation. The major cause of RA is asbestosis. RA is usually asymptomatic and may simulate a pulmonary neoplasm on chest Xray. The diagnosis is made by CT, demonstrating the pathognomonic "comet tail sign". No treatment is required. PMID- 16358663 TI - [Bleomycin induced flagellate erythema]. AB - Bleomycin is an antibiotic with antineoplastic pro prieties. It is used in the traitement of many syndromes. The cutaneous toxicity of bleomycin includes many alteration. We report a case of flagellate erythema following the administration of bleomycin in a 57-year-old women with Kaposi sarcoma. She developped linear lesions 6 days after the first dose of bleomycin. Flagellate erythema is a specific reaction to bleomycin therapy wich occurs in susceptible individuals independently of dose, route of administration and type of malignant disease treated. Its occurs in 10 to 35% of the treated patients. PMID- 16358664 TI - [Polyomavirus BK infection in renal transplant recipients]. AB - Beside acute rejection or immunosuppressive therapy toxicity, infection by Polyomavirus BK, usually not aggressive in immunoactive patients, has emerged as an important factor affecting graft function in renal transplant recipients. Indeed, one of the most important complications of BK infection is nephropathy. Viral replication in the urinary tract as assessed by the presence of "decoy cells", or by a positive PCR for BK virus has been detected in up to half of the recipients but only 5% will present nephropathy which is usually the only sign. The most common risk factors for this emerging new cause are new immunosuppressive drugs and rejection episodes. The gold standard to diagnose BK nephropathy is immunohistochemical staining for large T antigen in graft biopsy specimens. Urine cytology examination and DNA BK PCR are used as a screening test. The prognosis in BK nephropathy has been considered to be poor. The early reduction of immunosuppression can improve the prognosis and perhaps also cidofovir or leflunomide use. PMID- 16358665 TI - [Ankle sprain. Management of recent lesions and prevention of secondary instability]. AB - Ankle sprains are among the most common musculoskeletal injuries. The vast majority of these injuries are lateral ligament sprains, resulting from inversion of the plantar-flexed foot. Initial management should include rest, ice, compression and elevation. Functional treatment proved to provide the quickest recovery and an earlier return to work without compromising the lateral mechanical stability. Surgical management of acute ankle sprains is rarely, if ever, indicated. Patients who continue to sustain multiple recurrent inversion sprains despite a supervised rehabilitation program aimed at improving proprioception and strengthening of the peroneal muscles are candidate for surgical reconstruction of the lateral ankle ligaments. PMID- 16358666 TI - [Control of gene transcription as a new therapeutic target in asthma]. AB - The recent advances in the knowledge of the molecular mechanisms underlying asthma have lead to a significant improvement of the current treatments of the disease and opened new perspectives for the development of therapeutic alternatives to inhaled corticosteroids. The selective targeting of transcription factors controlling the expression of the genes implicated in the pathogenesis of asthma is one of these privileged strategies. This review aims at describing the most promising new therapeutic targets in the control of asthmatic inflammation at the gene transcription level. PMID- 16358667 TI - [Spotlight on sensory irritation and its treatment]. AB - Sensory irritation is directly bound to the concept of sensitive skin. A large proportion of the population, in particular young women, is affected. The reported symptoms are those of skin discomfort without any visible alteration of the skin. According to the subjects, the timing and the cutaneous sites, various perceptions occur, including pruritus, burning and stinging sensations. Sensory irritation is difficult to quantify because it remains subjective by essence. The condition is sometimes aggravated by some cosmetics, cleaning and cleansing products, wool contact and exposure to cold and dry climate. The responsible ingredients are often agents affecting functions of stratum corneum without being obvious chemical irritants. PMID- 16358668 TI - [Exogenous lipoid pneumonia: a simple diagnosis?]. AB - Exogenous lipoid pneumonia is a poorly known pathology. It results from aspiration of mineral, vegetal or animal oily substances. It is usually iatrogenic and clinical presentation is not specific. Main investigations which allow the diagnosis are: tomodensitometry which is the reference diagnosis modality, chest X-Ray which may be used for follow-up and MR which may contribute to the diagnosis. The treatment is not systematized. Stopping intake of the causal agent is essential. Some patients are treated with success by broncho alveolar lavage, corticotherapy and oxygenotherapy. An exogenous lipoid pneumonia must be considered in all cases of non resolving pneumonia. PMID- 16358669 TI - [How I explore. . . vasculitis of the peripheral nervous system]. AB - Vasculitic neuropathies require early diagnosis and therapy since their prognosis is potentially severe. The following features are usually suggestive of a vasculitic neuropathy: painful multiple mononeuropathy with an acute/subacute course, signs and symptoms of multisystem involvement (arthritis, asthma, renal and skin involvement), biological evidence of an inflammatory syndrome. In one third of patients however, vasculitic neuropathies present as a polyneuropathy. For other patients, peripheral neuropathy is the presentation mode of a necrotizing vasculitis, in the absence of biological inflammatory syndrome. A combined muscle and nerve biopsy, will usually demonstrate the vasculitic process. Vasculitic neuropathies often have a favourable prognosis when immunosuppressive therapy (corticosteroids and/or cyclophosphamide) is initiated at an early stage. PMID- 16358670 TI - [Analysis of the results of the main randomized trials on carotid endarterectomy for asymptomatic internal carotid stenosis]. AB - The authors analyse the ACAS (1997-93) and ACST (1993-2003) trials, comparing the best medical therapy vs endarterectomy of an asymptomatic 60 % or more stenosis of the internal carotid artery. The ACAS trial does not reveal any benefit of surgery in preventing major stroke during follow-up (stroke rate 3.4% vs 6.02% in medical group, N.S.). Only by considering minor and major strokes together, a beneficial effect of carotid endarterectomy could be evidenced (5.9% stroke reduction at 5 years: 5.1% ipsilateral stroke at 5 years after surgery vs 11% stroke at 5 years under medical treatment). Indeed, following surgery, the stroke risk (ipsilateral) at 5 years attained 5,1% vs 11% following medical treatment of an asymptomatic internal carotid artery stenosis of 60-99%. The ACST trial, set up in 1993, aimed to assess the long-term result of carotid endarterectomy for asymptomatic 60-99% stenosis. Patients randomized for immediate surgery had a slightly more favorable outcome than those allocated to medical treatment. Absolute risk reduction of major stroke at 5 years was 2.5%. Overall, the benefit of operating an asymptomatic carotid artery stenosis is rather low (1% less stroke per year and less than 0.5% major stroke reduction per year). This positive result is obtained only if the operative stroke-death rate remains extremely low (<2.8%). The indication for operating asymptomatic carotid stenosis must be cautiously taken; age of the patient, degree of stenosis and surgeon's experience and results are to be considered. PMID- 16358671 TI - [Insulin detemir (Levemir)]. AB - Insulin detemir (Levemir) is a soluble long acting human insulin analogue acylated with a 14-carbon fatty acid. The fatty acid modification allows insulin detemir to reversibly bind to albumin, thereby providing slow absorption and a prolonged metabolic effect (up to 24 hours) with low variability. Indeed, in patients with type 1 or type 2 diabetes mellitus, insulin detemir has a more predictable, protracted and consistent effect, with less intrapatient variability in glycaemic control (particularly fasting plasma glucose levels), compared with NPH (Neutral Protamine Hagedorn) insulin. Insulin detemir, is at least as effective as NPH insulin in maintaining overall glycaemic control, with a lower risk of nocturnal hypoglycaemia. It also provides the additional benefit of less body weight gain as compared to other basal insulins. Levemir, presented in cartridges for the pen device NovoPen 3 and administered preferably at bedtime (if necessary morning and evening), is a promising new option for basal insulin therapy in diabetic patients, especially those on a basal-bolus scheme. PMID- 16358672 TI - [First evidence of greater cardiovascular protective effects of newer as compared to old antihypertensive drugs treatments: the ASCOT-BPLA results]. AB - In the antihypertensive population, a normalisation of blood pressure level must be the first objective. The Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT) - Blood Pressure lowering arm again confirms this assertion. The benefit has been proportional to the blood pressure decrease. However, the use of newest antihypertensive drugs (the long acting dihydropyridine calcium antagonist amlodipine very often associated to the ACE inhibitor perindopril) in the hypertensive population aged from 40 to 79 years with at least 3 additional risk factors showed significantly lower rates of cardiovascular and total mortalities, cardiovascular events, new diabetes or renal impairment as compared to the atenolol-based combination drug regimen (with thiazide as required). It should be noted that almost all the patients also received a low dose of a hypolipidemic drug, atorvastatin. Both arms (the statin arm and the arm using antihypertensive drugs) have yet been stopped, after 3 and 5 years, respectively related to the large beneficial effects noted with the new antihypertensive and statin treatments, although the primary endpoints (non fatal myocardial infarctival and fatal coronary heart disease) were not reached. This study raises many unsolved questions about the explanation for such benefits or the future management of the hypertensive people whose characteristics were very similar to those people frequently encountered in clinical practice. However, the present study emphasizes the considerable efficacy of a combination of amlodipine-perindopril and a low dose of statin. PMID- 16358673 TI - Oral cancer. PMID- 16358674 TI - Tooth whitening agents --a legal maze. PMID- 16358675 TI - The president invites you! PMID- 16358676 TI - Treatment of third molar teeth assessment. AB - This first article will discuss the indications for removal of third molars and the initial assessment of the patient. The importance of good communication will be highlighted. PMID- 16358677 TI - Immediate Osseotite implant placement and immediate loading of a provisional restoration of maxillary lateral incisors. AB - This article describes the immediate replacement of two maxillary lateral incisors teeth after extraction of the left lateral deciduous incisor at the same time with immediate Osseotite implants and immediate restoration. A traumatic dental extraction of a deciduos (b) lateral incisor was performed and a 4mm diameter x 15 mm 3i tapered Osseotite (Implants Innovations, Palm Beach, CA, USA) implant was immediately placed. The other lateral incisor place was treated at the same time and provisional temporary crowns were placed at the same surgery. The provisional crowns did not have any occlusal contact to reduce negative lateral forces. Final impression for definitive restoration was made five months after implant placement veneer ceramic crowns were bonded to the Gingihue abutment (3i, Implants Innovations, Palm Beach, CA) one month later. This article describes the use of immediate implants with immediate loading of Osseotite combined with provisional crowns resulted in an excellent outcome after a two year follow up period. PMID- 16358678 TI - Identification periodontal risk factors prior to restorative treatment. PMID- 16358679 TI - Face the challenges. PMID- 16358680 TI - Dental project Uganda. PMID- 16358681 TI - Women in Irish dentistry. AB - In 1928, women constituted barely five per cent of Irish dentists. This can be attributed to a variety of factors, especially societal attitudes. However, there were perceptions that women brought certain attributes to the profession. By 1960, the proportion of women in dentistry had increased to 11%, and, by 1980, one-in-five dentists registered in the Republic of Ireland were female. PMID- 16358682 TI - Your practice is not your pension fund. AB - I believe that in the not too distant future--in 10 to 20 years--the single handed practice as we know it today will cease to exist. Even in today's market, 25% of practices have either closed due to an unsaleable commodity or have taken two to three years to sell and at a seriously reduced price. This thought alone should refocus the mind as we continue into the 21st century. PMID- 16358683 TI - Helping consumers understand dental fees. PMID- 16358684 TI - [Simultaneous determination of isomers of dihydroxybenzene using alternating trilinear decomposition algorithm combined with reversed-phase high performance liquid chromatography/diode array detection]. AB - Alternating trilinear decomposition (ATLD) algorithm is a promising method for second-order calibration, which is based on an alternating least-squares principle to overcome some shortcomings of the traditional parallel factor analysis (PARAFAC) algorithms, such as the sensitivity to the estimated component numbers and slow convergence. It utilizes an iterative procedure involving Moore Penrose generalized inverse computations based on singular value decomposition and can be applied to the simultaneous determination of several organic components even in the presence of unknown interfering components. The overlapped chromatogram and spectra system of dihydroxybenzenes were resolved by ATLD combined with reversed-phase high performance liquid chromatography (RP-HPLC)/ diode array detection (DAD). The elution time was set from 1.086 min to 1.399 min with an interval of 1/150 min and the ultraviolet wavelength from 268 nm to 298 nm with an interval of 1 nm. The resolved results were in agreement with the actual results excellently. Catechol (CAT), resorcinol (RES) and hydroquinone (HYD) in aqueous solution were determined simultaneously, whose recoveries were (100.1 +/- 1.0)%, (99.4 +/- 1.4)% and (100.5 +/- 1.7)%, respectively. The results showed this experiment can be easily performed without resorting to time consuming and complicated procedures and quantified accurately. It successfully solved the resolving problem caused by chromatogram and spectra overlapping in the presence of interfering components. It also revealed the ATLD algorithm can be efficiently applied to solve the second-order calibration problem in HPLC-DAD. PMID- 16358685 TI - [Determination of organophosphorus pesticides in vegetables by microwave-assisted extraction/gas chromatography-mass spectrometry]. AB - A method of microwave-assisted extraction (MAE) followed by gas chromatography mass spectrometry (GC-MS) for the determination of organophosphorus pesticides (OPPs) in vegetables has been developed. The OPPs studied were diazinon, parathion and isocarbophos. Several extraction solvents were compared on the MAE efficiency. Dichloromethane gave good recovery, and was chosenfor the procedure. A three-level orthogonal array design was used to optimize the MAE process.Factors affecting the MAE efficiency were considered, including the solvent amount and extraction time.The linear ranges of the method were from 4 ng/g to 400 ng/g for diazinon and parathion, and from 20 ng/g to 400 ng/g for isocarbophos. The detection limits were 0.29 ng/g, 1.70 ng/g and 2.30 ng/g for diazinon, parathion and isocarbophos respectively. Two sets of spiked vegetable samples of 200.0 ng/g and 50.0 ng/g were determined. The recoveries were from 72.2% to 102.0%, and the RSDs were from 1.5% to 11.0% (n = 3). The analytical results agreed quite well with those obtained by conventional extraction with mechanical shaking (EMS). The method is fast and solvent-saving. PMID- 16358686 TI - [Analysis of 7-aminonitrazepam in urine by trimethylsilyl derivatization-gas chromatography/mass spectrometry]. AB - A highly sensitive method has been developed for the analysis of 7 aminonitrazepam (7-ANIZ), the major metabolite of nitrazepam, in urine by trimethylsilyl derivatization-gas chromatography/mass spectrometry. Urine samples were extracted with ethyl ether-ethyl acetate (99:1, volume ratio). The extracts were derivatized with N,O-bis (trimethylsilyl) trifluoroacetamide, and the total ion current chromatograms of derivatives were acquired. 7-ANIZ was identified by the relative abundance of major characteristic ions in the mass spectrum of its derivative and the retention time of the mass chromatogram peaks of these characteristic ions. Based on the mass chromatogram of the base peak ion, quantification was performed using 7-aminoclonazepam (7-ACLZ) as the internal standard. The extraction efficiency of 7-ANIZ was 82.8%. The linear range was 10 microg/L - 500 microg/L. The limit of detection was 1.2 microg/L and the limit of quantification was 3.5 microg/L. The recoveries were 94.7% - 103.5%, and the RSDs were 3.9% - 5.4%. 7-ANIZ in the urine sample excreted by the subject over 96 h period after oral administration of 10 mg nitrazepam was measured. It is demonstrated that the method can be applied to the forensic identification. PMID- 16358687 TI - [Determination of amino acids and glucose in amino acid injection by anion exchange chromatography with integrated pulsed amperometric detection]. AB - A direct, sensitive, simple and practical method for the determination of amino acids and glucose in amino acid injection by anion exchange chromatography with integrated pulsed amperometric detection was developed. There is no need for sample pretreatment. The retention behavior of amino acids and glucose on the anion exchange column was investigated using sodium hydroxide and sodium acetate as eluent. When the concentration of sodium hydroxide was changed, the change in the retention times was generally greater for amino acids than for glucose. The difference can be used to improve the separation between amino acids and glucose. When the concentration of sodium acetate was changed, the change in the retention times was generally greater for amino acids containing two carboxyl groups than for amino acids containing only one carboxyl group. The difference can be used to improve the separation between di-carboxylic amino acids and mono-carboxylic amino acids. In order to separate amino acids and glucose, a ternary gradient elution method with water, sodium hydroxide and sodium acetate as mobile phases was employed. The optimized gradient elution condition for the analysis of 17 amino acids and glucose in amino acid injection was obtained. The time of the gradient elution program was 74 min. Under the optimized conditions and the column temperature of 30 degrees C, the detection limits for 17 amino acids and glucose were 0.3 pmol- 10.3 pmol. The calibration graphs of peak area for all the analytes were linear in the ranges about two orders of magnitude. The relative standard deviations (RSDs) (n = 5) were 0.7% - 3.8%. The recoveries of added standard were 88.3% - 104.6%. PMID- 16358688 TI - [Preparation of low electroosmotic flow monolithic capillary column and the investigation on its properties]. AB - Monolithic columns of capillary electrochromatography (CEC) with low electroosmotic flow (EOF) have been prepared by in-situ polymerization of butylmethacrylate and ethylene dimethacrylate, without any charged groups existed in the reaction mixture. The good reproducibility of the columns has been proved no matter if they are prepared in the same or different batches. Besides the traditional ternary porogenic mixture of 1-propanol, 1,4-butanediol and water, a binary porogenic mixture with only alcohols has also been adopted. In comparing with the ternary porogenic mixture, the design of binary mixture allows fine control of pore diameter and the formation of specific surface of the monolithic polymers. The effects of composition and ratio of porogenic reagents on EOF of the column systems have also been shown. The pore properties and EOF of monolithic columns with different porogenic mixtures were investigated. The results of scanning-electron micrograph show that the continuous beds were quite different from each other, while the proportions of 1-propanol were the same under these two situations. In addition, the Joule heat effect of such columns has been studied by varying the inner diameter of capillary columns. The effect of length of capillary on the EOF was also studied. Under the conditions of high pH and high concentrations of organic solvents, low EOF was obtained to satisfy different needs from the diversified separation modes. Through the separation of acidic compounds, monolithic columns with low EOF have shown potentialities in the analysis of charged samples. PMID- 16358689 TI - [Capillary electrophoretic enantioseparation of arylglycine amides with highly sulfated beta-cyclodextrin as chiral selector]. AB - Enantiomers of nine arylglycine amides synthesized were successfully separated by capillary electrophoresis (CE) using highly sulfated beta-cyclodextrin (HS-beta CD) as a chiral selector. Baseline enantioseparation of the analytes was obtained within 6 min at neutral pH but not the commonly used acidic condition. HS-beta-CD content, buffer type and concentration, and non-chiral additive were studied and optimized for high resolution and fast speed. A reproducible running buffer system composed of 15 g/L HS-beta-CD, 0 or 10% (volume fraction) methanol and 20 mmol/L 3-(N-morpholino) propane sulfonic acid at pH 6.5 was obtained. The D enantiomer always migrated ahead of the L-enantiomer for all the 9 pairs of arylglycine amides. The migration order was found to be dependent on the structure of the solutes. PMID- 16358690 TI - [Measurement of diffusion coefficients of polar solvent and nonpolar solvent at infinite dilution in polyethylene]. AB - Gas chromatography is a new, fast, accurate and convenient technique to study the correlation of small molecule solvents and polymer membrane materials. It can measure many parameters of dissolution and diffusing characters of a small molecule in a polymer. The retention times and the peak widths at half-height of five small molecule solvents (n-hexane, n-heptane, n-decane, ethanol and water) in the stationary phase of polyethylene were measured. The diffusion coefficients of the small molecule solvents at infinite dilution were calculated with van Deemter equation. The graph plotted according to the results of the diffusion coefficients of n-decane at infinite dilution vs temperatures agreed with the Arrhenius equation. The variance in the diffusion coefficients at infinite dilution of five small molecule solvents was analyzed according to the differences in molecular mass and polarity. PMID- 16358691 TI - [Studies on quantitative structure-retention relationships for sulfides on stationary phases of different polarities]. AB - Multiple linear regression (MLR) based on forward stepwise multiple regression techniques was used for predicting gas chromatographic retention index of sulfides after calculating their quantum-chemical parameters. The correlative quantitative model between the retention index (IR) of sulfides on four stationary phases of different polarities and their quantum chemical parameters was built. The results proved the strong predictive power of the models. The root mean-square error (RMS) of the regression equation was less than 4%. PMID- 16358692 TI - Study of isothermal adsorption of albumin on acetate fiber filter rod with dye affinity ligand. AB - Cibacron Blue F3GA was immobilized onto an acetate fiber filter rod to produce potential affinity matrix. The adsorption isotherms of the prepared matrix for either bovine serum albumin or human serum albumin were investigated. It was experimentally shown that the affinity isothermal adsorption obeyed Freundlich adsorption model. By means of affinity chromatography of human plasma using the prepared matrix, purified human serum albumin was obtained. PMID- 16358693 TI - [Improvement of prediction model of retention values of anions with sodium carbonate-sodium bicarbonate as eluents]. AB - A prediction model of retention values of anions with sodium carbonate-sodium bicarbonate as eluents has been improved. The improved model is suitable for a wider range of concentrations of sodium carbonate-sodium bicarbonate eluents than the old one set up several years ago. An extensive set of experimental retention data obtained for 7 anions (nitrate, sulfate, oxalate, selenate, iodide, phosphate and arsenate ) using the eluents of varied concentrations was dealt by binary linear regression process. The improved model gave satisfactory performance with correlation coefficients over 0.999 for nitrate, sulfate, oxalate, selenate and iodide, of 0.9971 for phosphate, and of 0.995 7 for arsenate. A computer program has been designed with the prediction model to simulate the separation of anions and used to optimize the routine analysis with sodium carbonate-sodium bicarbonate as the eluent. PMID- 16358694 TI - [Study of the separation factors of basic compounds in HPLC using a reversed phase amino-type column]. AB - The chromatographic behaviors of some basic probes and a neutral reference compound were investigated as functions of temperature, buffer cation strength and flow-rate using a novel reversed-phase amino-type column at both pH 3.0 and pH 7.0. The results showed that the retention of basic compounds declined steeply at first, but then slowly when temperature increased continually at pH 3.0 while decreased slowly in the whole range of temperature at pH 7.0. The column efficiency increased and the peak shape improved with increased temperature. The buffer cation strength has a strong effect on the retention of bases. The optimum flow-rate for the highest efficiency is generally lower for basic compounds than neutral compounds, and due to the steepness of the van Deemter curves obtained, high flow-rates appear to be particularly detrimental in the chromatography of basic compounds. All the results indicated that the retention mechanism, was more complex for the analysis of bases on amino-type columns than on traditional reversed-phase columns because of the existence of amino group in the alkyl group of amino-type columns. PMID- 16358695 TI - [Preparation of polymer modified stationary phases through surface radical chain transfer reaction]. AB - It is demonstrated that radical chain transfer to thiol-terminated self-assembled monolayer and surface-initiated polymerization could be used for in-situ grafting ultra-thin polymer film on silica. The widely used silane coupling agent mercaptopropyltrimethoxysilane (MPS) was used to prepare the thiol-terminated silica, and in the presence of which, chain-transfer reaction and polymerization of methyl methacrylate (MMA) was carried out using azo di-isobutylnitrile (AIBN) as the initiator. Both thiol-terminated silica and polymer film modified silica were characterized by Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, thermogravimetric analysis (TGA) and elemental analysis. The results showed that the grafting amount of polymer on silica was higher than the traditional methods, which can effectively relieve the non-specific adsorption of solute with uncovered hydroxyl groups on the surfaces of silica in subsequent chromatographic application. The obtained polymethylmethacrylate (PMMA) modified silica exhibited excellent separation capacity on the separation of the polar compounds and polycyclic aromatic hydrocarbons. This kind of surface initiated polymerization prospects for the preparation of polymer modified stationary phases. PMID- 16358696 TI - [Preparation of molecularly imprinted chiral monolithic column and its applications for separation of diastereomers]. AB - A cinchonine imprinted chiral monolithic column was prepared for the separation of the diastereomers of cinchonine and cinchonidine by in-situ molecular imprinting technique. This type of molecularly imprinted column can be prepared only by a single-step procedure. In order to improve the selectivity and efficiency, a new in-situ molecular imprinting polymerization system was introduced by adopting porogenic solvents of toluene and 1-dodecanol with relatively low polarity and appropriate ratios of polymerization mixture. Diastereomers of cinchonine and cinchonidine were fully separated both under isocratic and gradient elutions on the chiral monolithic column. The broad peak shown in isocratic elution could be improved in gradient elution. Effects of mobile phase composition, flow rate and temperature on retention and separation factor were investigated. Due to the large throughput pores in the chiral monolithic column, low backpressure was observed during the separation process and a separation factor of 3.18 was obtained at 1.0 mL/min. The increase of temperature could improve the separation factor and a maximum separation factor of 5.40 was obtained at 60 degrees C. PMID- 16358697 TI - [Rapid separation and determination of enterotoxigenic E. coli by capillary zone electrophoresis]. AB - Rapid separation and determination of enterotoxigenic E. coli K88, K99 and 987P with intact cells using capillary zone electrophoresis (CZE) are described. The CZE was performed in a running buffer of 0.05 mol/L Na2CO3-NaHCO3 (pH 9.9), under applied voltage of 14.1 kV and detected by an on-line UV at 210 nm. Results show that K88, K99 and 987P were separated completely. Moreover, each strain presented a characteristic peak and a reproducible retention time (RSD < or = 0.9%). The successful analysis of enterotoxigenic E. coli in the excrement of diarrhea piglets is also discussed. K88 and K99 were detected in the excrement of diarrhea piglets born in 5 d - 6 d, and 30 d - 35 d respectively, and nothing was detected in the excrement of diarrhea piglets born in about 60 d. PMID- 16358698 TI - [Determination of isoniazid in blood and urine samples by reversed-phase high performance liquid chromatography]. AB - For the purpose of more accurate and rapid analysis of isoniazid in body fluid samples, a reversed-phase high performance liquid chromatographic (HPLC) method was developed. Vanillin, being as a derivatizing reagent, was added to the plasma and urine samples when they were pretreated. Isoniazid and vanillin reacted to form isonicotinoyl hydrazone which was separated and detected. The pretreatment method of sample, the linear range, the precision, and the recovery of isoniazid were investigated by using human's plasma and urine spiked with standard isoniazid. The linear range was 0.2 mg/L- 12.0 mg/L (for plasma, r =0.999 6; for urine, r = 0.999 4). The detection limit was 0.2 mg/L. The intra-day and inter day precisions of assay for isoniazid were 2.3% - 2.6% and 3.8% - 4.0% (n = 5) for plasma, 1.3% - 4.0% and 2.1% - 3.9% (n = 5) for urine respectively. The average recoveries of isoniazid were from 96.5% to 99.8% in plasma and from 96.3% to 99.4% in urine. The HPLC method has been used to investigate the concentration of isoniazid in the volunteer's plasma. The quantitative analysis of isoniazid in plasma was not interfered by impurities, the metabolites of isoniazid and the derivatizing reagent residue. This analytical method for isoniazid is sensitive, rapid and convenient. It is suitable for the analysis of toxics in forensic samples, the detection of drug concentration in clinical medicines and pharmacokinetic studies. PMID- 16358699 TI - [Determination of andrographolide and dehydroandrographolide in Andrographis paniculata nees materials and related patent medicines by reversed-phase high performance liquid chromatography]. AB - A simple and accurate method for the determination of andrographolide and dehydroandrographolide in andrographis paniculata Nees materials and patent medicines with high performance liquid chromatography (HPLC) has been developed. The two components were extracted from powdered samples by shaking with methanol. The resultant extracts were separated within 15 min on a BECKMAN C18 column (4.6 mm i. d. x 250 mm, 5 microm) and with a gradient elution of acetonitrile-water at a flow rate of 0.5 mL/min. The detection wavelength was 225 nm and the injection volume was 20 microL. In gradient elution program the volume fraction of acetonitrile in mobile phase was as follows: 0 min - 1 min, 40%; 1 min - 5 min, 40% - 50%; 5 min - 15 min, 50% - 70%. Both andrographolide and dehydroandrographolide have good linearity in the range of 10 mg/L to 100 mg/L with the correlation coefficients of 0.997 6 and 0.998 6 respectively. This method has been successfully applied for the analysis of andrographis paniculata Nees materials and related patent medicines. PMID- 16358700 TI - [Determination of cinnabar in zijinhong capsules by high performance liquid chromatography]. AB - A high performance liquid chromatographic method was established for the determination of cinnabar in Zijinhong capsules using diethyldithiocarbamate (DEDTC) chelate. A Waters X-Terra C18 column was used with a mixture of methanol 0.01 mol/L Na2HPO4 (pH 7.5) (containing 0.01% DEDTC) (73:27, V/V) as the mobile phase and a detector set at 270 nm. The calibration curve was linear (r = 0. 999 6) in the range of 10.1 mg/L - 100.9 mg/L of Hg2+. The recoveries were 97.0% - 100.8% with RSDs of 1.8% - 2.3%. PMID- 16358701 TI - [Determination of anions in the products of ozonation of reactive dye by ion chromatography]. AB - A method for the determination of anions in the degradation products of C. I. Reactive Red 120 by ozonation was investigated. The sample can be pretreated with a Dionex OnGuard P column, which has high selectivity for removing phenols, azo group-contaning compounds, aromatic carboxylic aicds etc. For the separation of organic and inorganic anions in the products, the anion exchange chromatograhy with gradient elution of NaOH was used. All the species were detected by both suppressed conductivity and UV absorbance detectors. The sulfate, oxalate, chloride, nitrate and formate could be identified even in very low concentration with ion chromatography, within 18 min, with the recoveries between 91.6% - 108.3%. The results indicate that the method is reliable, simple, rapid and especially sensitive. In combining with the determination results of conventional parameters of the solution of Reactive Red 120, a preliminary elucidation of the degradation mechanism PMID- 16358702 TI - [Determination of 2-methyl-3-nitrobenzoic acid through pretreatment with diazomethane by gas chromatography]. AB - A method for the quantitative determination of 2-methyl-3-nitrobenzoic acid by gas chromatography is described. 2-Methyl-3-nitrobenzoic acid was esterified by pretreatment with diazomethane prior to analysis. A CP-Sil-43CB capillary column(25 m x 0.32 mm i.d. x 0.2 microm), a flame ionization detector and the area normalization method were used. The average recovery was 99.81%. The RSD was 0.08% and the detection limit was 3 x 10(-11) g. The results showed that the method is practical and reliable. It was realized that the higher purity and higher boiler matter was analyzed by gas chromatography. The method can be used to monitor the purity of this type of materials. analysis of research and production. It can be used in the development of new products and in the process. PMID- 16358703 TI - [Determination of the conversion of air oxidation of penicillin derivatives by high performance liquid chromatography]. AB - In air oxidation of penicillin G p-methoxybenzyl ester (PGPMB) to its sulfoxides (PGPMBO), sol-gel technique was employed to encapsulate the catalyst Co (acac)3, by which the reaction was run under heterogeneous conditions. A reversed-phase high performance liquid chromatographic method was established for the determination of the conversion of this reaction. The analysis of PGPMBO was carried out on a C18 column (4.6 mm i.d. x 150 mm, 10 microm) with the mobile phase of methanol-water (85:15, V/V), at a flow rate of 1 mL min(-1) and with detection wavelength of 270 nm. The linearity of the calibration curve of PGPMBO was obtained over the range of 0.05 g x L(-1) - 0.40 g x L(-1) with a relative coefficient of 0.999 2. The highest conversion was 98.8%. The method can be applied to determine this conversion promptly in the reaction system. PMID- 16358704 TI - [Determination of oxolinic acid residues in baked and fresh eels by high performance liquid chromatography without derivatization]. AB - A method for rapid determination of oxolinic acid residues in baked eels and fresh eels by high performance liquid chromatography without derivatization was studied. The samples were triturated, and extracted with methylene dichloride. The extracted solutions were evaporated to dryness, and then was dissolved by using the mobil phase. The solutions were cleaned-up by partition with n-hexane before analysis. The components were analysed on a Nova-Pak C18 column (150 mm x 3.9 mm i.d., 5 microm), with methanol-diluted phosphoric acid (pH 2.5) (40:60, V/V) as the mobile phose at a flow rate of 0.8 mL/min and detected by a fluorescence detector at the excitation wavelength (lambdaex) 254 nm and the emission wavelength (lambdaem) 375 nm. The detection limit was 0.01 mg/kg (S/N = 3). The recoveries were over 80 %, and the relative standard deviations (RSDs) were less than 3.8%. PMID- 16358705 TI - [Determination of benzoyl peroxide in wheat flour by high performance liquid chromatography]. AB - An improved method for the determination of benzoyl peroxide in wheat flour is reported. Benzoyl peroxide was extracted with petroleum ether, and then analysed by reversed-phase high performance liquid chromatography on a Shim-Pack VP-ODS column (0.46 cm i.d. x 15 cm, 5 microm - 6 microm) using a mobile phase of acetonitrile-0.3% H3PO4 aqueous solution (4:1 in volume ratio) with a flow rate of 1.0 mL/min, and UV-236 nm detection. A linear correlation was observed between 0.002 g/L- 0.012 g/L of benzoyl peroxide. The detection limit was 0.005 g/kg. This proposed method without redox reaction is superior to colorimetric determination. This method is characterized with simplicity, convenience, reproducibility and accuracy. PMID- 16358706 TI - [Analysis of volatile constituents of root and rhizome of Asarum heterotropoides Fr. var. mandshuricum (Maxim.) Kitag. by gas chromatography-mass spectrometry]. AB - The volatiles of root and rhizome of Asarum heterotropoides Fr. var. mandshuricum (Maxzim. ) Kitag. were extracted with n-hexane and analyzed by capillary gas chromatography-flame ionization detection and gas chromatography-mass spectrometry (GC-MS) with the same column HP-5 (30 m x 0.25 mm i.d.). Initial oven temperature was kept at 40 degrees C for 2 min, then raised to 160 degrees C at 3 degrees C /min, immediately raised to 250 degrees C at 6 degrees C /min, and kept 250 degrees C for 23 min. According to GC-MS analysis, 36 compounds were identified. The main components are safrole, methyleugenol, 3,5-dimethoxytoluene, pentadecane, N-isobutyldodecatetraenamide. Furthermore, the analysis results of root and rhizome were compared. The results indicate that the fundamental constituents are nearly the same, but the quantities of volatiles in root are richer than those in rhizome. In addition, 8 compounds are selected as the criteria of quality control, including 3-carene, 1,8-cineole, eucarvone, methyleugenol, safrole, myristicin, pentadecane and N-isobutyldodecatetraenamide. PMID- 16358707 TI - [Determination of major flavor components in chinese spirits using a special domestic gas chromatograph with DNP packed column]. AB - A method using a domestic gas chromatograph controlled by computer with a homemade DNP packed column for the determination of major flavor components in Chinese spirtis is described. This chromatograph was developed under the sponsorship of the Ministry of Science and Technology. Optimizations of the operating parameters were undertaken in order to establish the chromatographic conditions suitable for isothermal and programmed temperature gas chromatography. The expanded analytical objects, including acetone, iso-valeraldehyde, 2 pentanone, 2-pentanol, acetoin, could also be quantitatively determined along with more than ten major components such as alcohols, esters, aldehydes, therefore, a total of 18 flavor components were accurately determined by this method. Under the selected chromatographic conditions, the instrument showed good stability and the repeatability of quantitative results was satisfactory. The suitability of this gas chromatograph is also discussed. PMID- 16358708 TI - [Determination of acetochlor and oxyfluorfen by capillary gas chromatography]. AB - A method is described for the determination of acetochlor and oxyfluorfen by capillary gas chromatography with FID and an SE-30 capillary column (60 m x 0.53 mm i. d., 1.5 microm), using dibutyl phthalate as the internal standard. The standard deviations for acetochlor and oxyfluorfen concentration(mass fraction) were 0.44% and 0.47% respectively. The relative standard deviations for acetochlor and oxyfluorfen were 0.79% and 0.88% and the average recoveries for acetochlor and oxyfluorfen were 99.3% and 101.1% respectively. The method is simple, rapid and accurate. PMID- 16358709 TI - [Analysis of sucrose esters by thin-layer chromatography]. AB - The thin-layer chromatographic method of sucrose esters was developed by the use of toluene-ethyl acetate-methanol-water (10:5:4.5:0.2 in volume ratio) as a mobile phase. Sucrose esters were developed by ascending technique on the silica gel G plate, visualized by urea-phosphoric acid-n-butanol with blue spots. The Rf value of monoester was 0.16 and those of higher esters were 0.38 - 0.93. The calibration curve was linear in the range of 25 microg - 250 microg with the detection limit of 0.1 microg. Accuracies were certified by normalization method and external standard method with t-test. The results were /t/ = 0.627(< 2.571) and /t/ = 1.123(<2.571) and RSDs were 3.03% and 3.08% (n = 6) respectively for S 1570. Contents of monoester of different sucrose esters were determined, which could provide reference for synthetic conditions and application range of sucrose esters. PMID- 16358710 TI - [Influence of consumables on performance of gas chromatography]. AB - The consumable materials of GC have a great influence on its performance. Usually, the materials are filters, column ferrules, septa, liners and so on. The points on their properties, how to choose them, how to install them and how they may affect the performance of GC are described. PMID- 16358711 TI - [Enhanced resistance to phytopathogenic bacteria in transgenic tobacco plants with synthetic gene of antimicrobial peptide cecropin P1]. AB - Plasmids with a synthetic gene of the mammalian antimicrobial peptide cecropin P1 (cecP1) controlled by the constitutive promoter 35S RNA of cauliflower mosaic virus were constructed. Agrobacterial transformation of tobacco plants was conducted using the obtained recombinant binary vector. The presence of gene cecP1 in the plant genome was confirmed by PCR. The expression of gene cecP1 in transgenic plants was shown by Northern blot analysis. The obtained transgenic plants exhibit enhanced resistance to phytopathogenic bacteria Pseudomonas syringae, P. marginata, and Erwinia carotovora. The ability of transgenic plants to express cecropin P1 was transmitted to the progeny. F1 and F2 plants had the normal phenotype (except for a changed coloration of flowers) and retained the ability to produce normal viable seeds upon self-pollination. Lines of F1 plants with Mendelian segregation of transgenic traits were selected. PMID- 16358712 TI - [Modification of the sunflower defensin SD2 gene sequence and its expression in bacterial and yeast cells]. AB - To achieve broader range of the defensin antimicrobial activity, based on the sd2 gene sequence, the modified gene, sd2mod, was constructed. Hybrid genes, sd2 licBM2, licBM2-sd2, licBM2-sd2mod, and sd2mod-licBM2, in which the wild-type and modified gene sequences were fused in frame with the reporter gene encoding thermostable lichenase, were constructed. Expression of the wild-type, modified, and hybrid genes was examined in the cells of pro- and eukaryotes. It was demonstrated that these genes were efficiently expressed in the cells of lower eukaryotes, the yeast. Inhibiting effect of the SD2 and SDmod proteins as the components of the hybrid proteins, SD2-LicBM2 and SD2mod-LicBM2, on the growth of the Fusarium culmorum hyphae was similar to that of the wild-type and modified proteins. It was shown that the presence of lichenase in the hybrid proteins facilitated selection and analysis of the hybrid proteins expression in transgenic organisms. PMID- 16358713 TI - [O6-benzylguanine stimulates regulatory functions of the Ada protein in Escherichia coli]. AB - In vitro experiments showed that O6-benzylguanine (O6-benzG, 0.2 microM) fully inhibited the repair activity of human O6-alkylguanine-DNA alkyltransferase (MGMT) due to the formation of S-benzylcytosine in the protein acceptor site. O6 benzG at concentrations increased many times (up to 800 microM) failed to inhibit the repair activity of the Escherichia coli Ada protein, the structural and functional analog of MGMT. It has been shown for the first time that O6-benzG stimulates the regulatory activity of the Ada protein. In experiments with N nitroso-N-methylurea (NMU), the pretreatment of Escherichia coli cells with O6 benzG at a sublethal concentration of 10 microM led to a twofold enhancement of transcription at the Ada-dependent promoter of the alkA gene in control cells and ensured transcription enhanced 1.6-1.7 times at alkA and alkB promoters in cells with the induced "classical" Ada response. Apparently, an increase in the regulatory activity of the Ada protein was associated with the formation of the stable protein molecule having the strong affinity for alkA and alkB promoters after transfer of the benzyl group from O6-benzG to the acceptor site Cys-69 in the N-terminal domain of Ada protein. O6-benzG did not affect the regulative activity of Ada in alternative quasi-adaptive responses to NMU. PMID- 16358714 TI - [The M/SAR elements of the bithorax complex in Drosophila melanogaster]. AB - The bithorax (BX) complex of Drosophila is a complex polygenic region with a multifactorial system of regulation. One of the levels of the regulatory system of the BX complex is its association with the nuclear skeleton structures through a specific interaction of the M/SAR DNA with the nuclear matrix proteins. In the present work, M/SAR elements were mapped on the molecular-genetic map of the region. All of the elements examined were found to colocalize with regulatory elements and form clusters that restrict/bracket the genetically active domains. All M/SAR DNA revealed was shown to bins specifically to the purified Drosophila melanogaster lamin. PMID- 16358715 TI - [The ecdysoneless1 gene regulates metabolism of the juvenile hormone and dopamine in Drosophila melanogaster]. AB - The dopamine (DA) content and the level of juvenile hormone (JH) degradation were studied in females of the wild-type Canton S strain and the ecdysoneless1 (ecd1) mutant, which does not produce ecdysone at a restrictive temperature (29 degrees C). Exposure at the restrictive temperature considerably increased the JH hydrolyzing activity and the DA content in five-day ecd1 females compared with flies of both strains growing at 19 degrees C and Canton S females exposed at 29 degrees C. In one-day ecd1 females, the level of JH degradation also increased at the restrictive temperature, but the DA content was low. The effect of ecdysone deficiency on the stress response in Drosophila melanogaster females was studied using changes in DA content and JH degradation were used as indices. The ecd1 mutation did not prevent the initiation of the stress response in females exposed at the restrictive temperature, but changed its intensity (stress reactivity). The interaction of 20-hydroxyecdysone with JH and DA in regulating Drosophila reproduction under normal conditions and in stress is discussed. PMID- 16358716 TI - [Genetic analysis of the predisposition to audiogenic seizure fits in Krushinsky Molodkina rat strain]. AB - The expression of audiogenic seizure fits has been studied in F1 hybrids between audiogenic seizure-prone Krushinsky-Molodkina rat strain and Wistar rats not prone to audiogenic seizures, as well as in two backcross generations. Only 10% of F1 hybrids exhibit audiogenic seizure fits, whereas the frequency of this character in two generations of their backcrosses with Krushinsky-Molodkina rats is about 50%. A digenic model with incomplete penetrance has been put forward to explain the control of audiogenic seizure fits. This model fits the data obtained: the theoretically expected distributions of the character in offsprings of different crosses do not differ significantly from those observed in experiments. The model explains why the distribution of the character is the same in the first and second backcross offsprings. PMID- 16358717 TI - [Estimating the gene pool condition in natural populations of invertebrates in the fragmented landscape of Moscow and Moscow district with special reference to shrub snail Bradybaena fruticum Mull]. AB - Using shrub snail Bradybaena fruticum Mull (20 populations) as a model, we were the first in Russia and in the world to develop a system of urban ecological genetic monitoring of the gene pool of an invertebrate species. The results of isozyme polymorphism studies in shrub snail populations showed a dramatic (up to 70%) reduction in genetic diversity in small isolates from the urbanized environment as compared to large natural populations. In urban populations, genetic diversity parameters were demonstrated to be lower than in natural ones: the mean heterozygosity per locus was reduced up to 0.08 (0.15-0.20 in natural populations); the mean allele number, to 1.9 (2.7 in natural populations); and the number of polymorphic loci, to four, i.e., 2.2-fold (nine in natural populations). In Moscow district, the number of polymorphic loci also decreased to five in the population subjected to anthropogenic pressure. The changes in the population gene pool (as shown by the number of polymorphic loci) were different in Moscow and Moscow district. The percentage of populations with the number of polymorphic loci as low as four to six was 76.9 in Moscow and 23.1% in Moscow district. The gene pool quality of 80% of the urban snail populations was estimated as unsatisfactory, and in half of them, as critical. The main reason for these changes seems to be genetic drift accompanied by inbreeding, caused by fragmentation of the range and reduction in the abundance of populations of the species, due to the anthropogenic pressure. The results of the study were employed in the program of the Moscow government for restoring the gene pools of endangered animals species on the preserved territories of the city. PMID- 16358718 TI - [Structural and transcriptional variation of mitochondrial DNA in pollen-sterile agamospermous sugar beet (Beta vulgaris L.) progeny]. AB - The cytoplasm status according to mitochondrial sequence tags was determined in agamospermous sugar beet progenies characterized by unstable manifestation of cytoplasmic male sterility. The detected variations in the ratios of homologous sequences related to the N and S mtDNA types did not correlate with pollen phenotypes of the plants. Polymerase chain reaction allowed semiquantitative evaluation of these variations and their detection. A cDNA corresponding to a hitherto unknown minor RNA and containing a consensus sequence of dicotyledonous plant promoters was detected by mtRNA display. Probable sources of CMS variability in agamospermous sugar beet progenies are discussed. PMID- 16358719 TI - [Development and study of SCAR markers in pea (Pisum sativum L.)]. AB - In order to develop more specific markers that characterize particular regions of the pea genome, the data on nucleotide sequences of RAPD fragments were used for choosing more extended primers, which may be helpful in amplifying a fragment corresponding to the particular DNA region. Of the 14 STS markers obtained from 14 polymorphic RAPD fragments, 12 were polymorphic, i.e., they are SCAR markers that can be used in genetic analysis. The transition from complex RAPD spectra to amplification of a particular SCAR marker substantially facilitates analysis of large samples for the presence or absence of the examined fragment. Inheritance of the developed SCAR markers was studied in F1 and F2. SCAR markers were used to identify various pea lines, cultivars, and mutants. It was established that the study of amplification of STS markers in various pea genotypes at varying temperatures of annealing and the comparison with amplification of the original RAPD fragments in the same genotypes provide an approach for analysis of RAPD polymorphism type. PMID- 16358720 TI - [Localization of genes controlling spherical grain and compact ear in Triticum antiquorum Heer ex Udacz]. AB - The inheritance of two taxonomically important characters was studied in hexaploid wheat species (2n = 6x = 42). The monogenic control of spherical grain was demonstrated for Triticum antiquorum Heer ex Udacz. The recessive gene controlling spherical grain in this species was assigned to chromosome 3D by monosomic genetic analysis and was shown to be allelic to the s gene determining the same character in the endemic Indian species T. sphaerococcum Perciv. The T. antiquorum and T. sphaerococcum dominant genes controlling compact ears proved to be nonallelic to the corresponding T. compactum Host. gene and were designated as C2. Problems of phylogeny and classification of hexaploid wheats are discussed. PMID- 16358721 TI - [Allozyme polymorphism of Swiss stone pine Pinus cembra L. in mountainous populations of the Alps and the Eastern Carpathians]. AB - Swiss stone pine Pinus cembra L. is a species with fragmented range, occurring in the Alpine-East Carpathian mountain system. Seeds of P. cembra are dispersed by nutcrackers, which offers potential possibilities for gene exchange among populations. Using isozyme analysis, we have examined five samples from two parts of the Swiss stone pine range: the Alps (Switzerland and Austria) and the Carpathians (two samples from the northern macroslope of the Gorgany Ridge, Eastern Carpathians, Ivano-Frankovsk oblast and one sample from Trans-Carpathian oblast of Ukraine). The allele frequencies of 30 isozyme loci, coding for enzymes ADH, FDH, FEST, GDH, GOT, IDH, LAP, MNR, MDH, PEPCA, 6PGD, PGI, PGM, SDH, SKDH, SOD, were analyzed using cluster analysis and methods of principal components. Two clusters, corresponding to the isolated Alpine and Carpathian parts of the range, were found. The main contribution to these differences were made by loci Adh-1, Adh-2, Fest-2, Lap-3, Mdh-4, and Sod-4. The interpopulation differentiation proved to be somewhat higher than that typical for pines (F(ST) = 7.4%), but within the limits characteristic for taxonomically close species. Thus, isolation of the populations did not lead to their marked differentiation, which may be explained by gene flow and balancing selection, which equalizes gene frequencies across the fragmented species area. Interlocus F(ST) heterogeneity (from 0.003 to 0.173) suggests adaptive significance of some of the allozyme polymorphisms or linkage of some loci with adaptive genes. The Carpathian populations were shown to have higher gene diversity than the Alpine ones (expected heterozygosities 0.095-0.114 and 0.060-0.080, respectively). A deficit of heterozygotes (as compared to the Hardy-Weinberg proportions), observed in the embryo sample, was probably explained by inbreeding. The reduction in the area of Carpathian pine forests in Holocene, caused by the global climatic changes and the anthropogenic impact, is hazardous for the gene pool of the species. The maintenance of genetic uniqueness of both Carpathian populations of P. cembra in general, and individual stands in particular, requires special measures for protection of Swiss stone pine in the Eastern Carpathians. PMID- 16358722 TI - [Inheritance of coat color in the mole vole (Ellobius talpinus Pallas)]. AB - Based on the ecological features of the mole vole, family analysis of the inheritance of coat color was performed with the use of material collected in a wild population. Analysis of coat color in parents and offspring has demonstrated that the offspring segregation into black and nonblack animals after crosses of different types agrees with the hypothesis on the monogenic inheritance of these color variations. Black mole voles are homozygous for the recessive allele (genotype aa). Homozygotes for the dominant allele (AA) are brown. Heterozygotes (Aa) may be brown or have transitional color. The mean frequency of brown coat color in heterozygotes is 0.509 and is very variable. The higher the color intensity in black elements of parent coat color, the more is the offspring coat color saturated with these elements. PMID- 16358723 TI - [Comparison CCR5de132 mutation in the CCR5 gene frequencies in Russians, Tuvinians, and in different groups of HIV-infected individuals]. AB - The 32-bp deletion (CCR5del32 mutation) in the CCR5 (chemokine (C-C motif) receptor 5) gene, encoding CCR5 chemokine receptor, is one of the factors determining natural resistance to human immunodeficiency virus (HIV-1) infection. In the present study, the samples of Russians (n = 107), Tuvinians (n = 50), and HIV-infected individuals were examined for the presence of CCR5del32 mutation in the CCR5 gene. The CCR5del32 allele frequency in Russians and Tuvinians constituted 7.84 and 2%, respectively. Among HIV-1 infected individuals, two groups, of macrophage-tropic HIV-1 strain- and T-cell-tropic HIV-1 strain infected were distinguished. The CCR5del32 allele frequency in the first group (6.45%) was lower than in the second one (8.73%). Statistical treatment of the HIV-1 infected individuals typing data showed that the difference in the CCR5del32 allele frequencies between the groups of sexually (macrophage-tropic) and parenterally (T-cell-tropic) infected individuals observed was within the limit of random deviation. PMID- 16358724 TI - [Polymorphism of alcohol dehydrogenase gene ADH1B in eastern Slavic and Iranian speaking populations]. AB - Frequencies of alleles and genotypes for alcohol dehydrogenase gene ADH1B (arg47his polymorphism), associated with alcohol tolerance/sensitivity, were determined. It was demonstrated that the frequency of allele ADH1B*47his, corresponding to atypical alcohol dehydrogenase variant in Russians, Ukrainians, Iranians, and mountain-dwellers of the Pamirs constituted 3, 7, 24, and 22%, respectively. The frequencies established were consistent with the allele frequency distribution pattern among the populations of Eurasia. Russians and Ukrainians were indistinguishable from other European populations relative to the frequency of allele ADH1B*47his, and consequently, relative to specific features of ethanol metabolic pathways. The data obtained provide refinement of the geographic pattern of ADH1B*47his frequency distribution in Eurasia. PMID- 16358726 TI - [Dynamic regimes in a model of single-locus density-dependent selection]. AB - A model of density-dependent selection in a Mendelian single-locus population was analyzed in the case where the fitnesses of genotypic forms are exponential functions of the population size. Analytical and numerical studies of the model were performed for a diallelic locus, and parametric regions were established for different dynamic behaviors of the model. The diallelic model of density dependent selection was generalized to a multiallelic locus; the results of its analysis are described. PMID- 16358725 TI - [Mutations and polymorphisms in the genes for myocilin and optineur in as the risk factors of primary open-angle glaucoma]. AB - A collection of DNA samples obtained from primary open-angle glaucoma (POAG) patients from St. Petersburg was analyzed for single-strand conformation polymorphism (SSCP) to reveal sequence variants in exon 3 of the myocilin gene (MYOC/TIGR) and in exons 4 and 5 of the optineurin gene (OPTN), where most of the mutations revealed worldwide are located. The Q368X mutation (c. 1102 C --> T) in exon 3 of MYOC/TIGR was detected in 1.2% (2/170) of the POAG patients from St. Petersburg, i.e., with the frequency close to that observed in other world populations. Three known polymorphisms in exon 3 of MYOC/TIGR, Y347Y (c. 1041 T - > C) (12.4%), T325T (c. 975 G --> A) (0.6%), and K398R (c. 1193 A --> G) (0.6%) were also detected. No statistically significant differences in frequencies of these polymorphisms were revealed between the POAG patient and control groups. The L41L polymorphism (c. 433 G --> A) in exon 4 of OPTN was detected in 2.9% of probands and in 1% of controls. The frequency of heterozygotes for the M98K polymorphism (c. 603 T --> A) in the OPTN exon 5 was statistically significantly higher (P = 0.036; Fisher's exact test) among the POAG patients (6.5%) than among the controls (1%). In the sample examined the E50K mutation, typical of the patients with pseudonormal intraocular pressure glaucoma, was not found. PMID- 16358727 TI - [Thermoanaerobacter ethanolicus gene cluster containing the alpha- and beta galactosidases genes melA and lacA, and properties of recombinant lacA]. AB - The nucleotide sequence of a 4936 bp Thermoanaerobacter ethanolicus genomic DNA fragment containing the thermostable beta-galactosidase gene lacA and two incomplete open reading frames has been determined. The product of the first frame is highly homologous to alpha-galactosidases (melibiases), the product of the third frame is homologous to the alpha-D-mannosidases. The terminal area of the lacA, immediately following the stop-codon, harbors presumably a transcription termination site. Based on the location of the putative alpha galactosidase gene melA and of the beta-galactosidase gene lacA on the T. ethanolicus chromosome, their combined transcription could be presumed. The calculated molecular mass of LacA is 86 kDa. LacA belongs to GH family 2 (GH2). Maximal activity of the purified recombinant enzyme was observed between pH values of 5.7 and 6.0 and temperatures of 75-80 degrees C. The highest activity, 480 units mg(-1), was found on lactose (Km 30 mM), the activities on pNPhGal and oNPhGal amounting to 330 and 420 units mg(-1), respectively. Immobilization on aldehyde silochrome increases the thermostability of the enzyme and keeps its high activity. PMID- 16358728 TI - [Detection of fluoroquinolone resistance SNPs in gyrA and parC genes of Neisseria gonorrhoeae using MALDI-TOF mass-spectrometry]. AB - For many known mechanisms of the drug resistance in microorganisms are described genetic markers (specific changes in the genome of microorganism, in the majority of the cases representing single nucleotide polymorphism). The search for the new methods, which make possible to identify single nucleotide changes with the greater effectiveness and at smaller prime is actual for the solution of the problem of the identification of the resistant strains. In this work a new approach of the determination of single nucleotide polymorphisms is proposed. It is based on the reactions of mini-sequencing and/or sequencing with the subsequent Matrix-Assisted Laser Desorption/Ionisation Time Of Flight Mass Spectrometry (MALDI-TOF MS) of the reaction products. The approach was tested on a clinical group of Neisseria gonorrhoeae strains to investigate specific single nucleotide polymorphisms in genes gyrA and parC (the genetic markers of the bacterium fluoroquinolone resistance). The results of the nucleotide polymorphism deter- mination was completely agreed with the data, obtained earlier with the use of a "gold standard" (sequencing with the classical gel-electrophoresis separation of the reaction products). There is specific interest in the method of sequencing of the short DNA sequences using MALDI-TOF MS. The new high-throughput approach of the single nucleotide polymorphisms determination in bacterial genes considerably increases the effectiveness of the methods of microorganism's identification, genotyping and determining the genetic markers of the drug resistance. PMID- 16358729 TI - [Identification of the rat ceruloplasmin mRNA isoform putative coded of a protein localized in mitochondria]. AB - Alternative expression of ceruloplasmin (Cp) gene, whose product, blue multicopper ferroxidase, is a neuron survival factor, was studied in the current work. Computer analysis showed that Cp-mRNA isoform, coding for 109 kDa polypeptide, can be formed as a result of the transcription from the alternative promoter in 3'-region of intron 2 of rat Cp gene. Alternative Cp form starts with 25 amino acid residues sequence, coded with intron 2 region. It is followed by amino acid sequence of the main Cp isoform starting from Gly 113. In silico data were experimentally confirmed using RT-PCR. It was demonstrated that the predicted mRNA was generally localized in liver and brain cells of adult rats. Direct sequencing of the obtained PCR-product showed the entire coincidence of the real and predicted mRNAs. It was in vitro showed that approximately 110 kDa Cp-like protein was completed and accumulated in the absence of mitochondria. This protein is transferred into the isolated mitochondria in the reconstructed system. Transport is energy-dependent, it is not accompanied with the shortening of Cp polypeptide length and needs the presence of cytosolic factors. Probably import is determined by the inner protein mitochondria import signal with amino acid sequence KVVYREFTDSTFRE, located in 756-769 region of mature Cp. Possible role of Cp in iron metabolism in mitochondria is under discussion. PMID- 16358730 TI - [Nucleotide correspondence between protein-coding sequences of Helicobacter pylori 26695 and J99 strains]. AB - Comparison of open-reading frames (ORFs) H. pylori 26695 and J99 strains has been revealed prevalence of nucleotide replacements as transitions (more than 3%) above transversions (less than 1%). Prevalence of nucleotide transitions is caused by high speed of C : G to T : A transitions in a coding strand of DNA (3.5 5.3%) and not coding strand (2.9-3.9%). The correspondence rate of transversion (A --> C, A --> T, C --> A, C --> G, G --> C, G --> T, T --> A and T --> G) did not exceed 0.84%. The highest correspondence frequency between C and T was detected in ACGT-ATGT (28.3%) - the site of methylation by active methyltransferase M.Hpy99XI in H. pylori 26695 and J99. Thus one can speculate that predominant transition taking place in H. pylori is mutation of C into T, which is realized through cytosine methylation-deamination mechanism. PMID- 16358731 TI - [Alternative promoters and oct-1 gene tissue-specific regulation]. AB - Murine oct-1 gene contains exons IU and 1L at 5'-end, which are alternatively presented in oct-1 mRNA. 1U exon is presented in oct-1 mRNA expressed in all tissues whereas lymphoid cells of mouse and human express mRNA containing alternatively IU or 1L exon. Upstream of these exons there are U and L promoters, respectively, which are separated by 67 kb in mice otf-1 locus. Nucleotide sequences located upstream 1U and 1L exons are very different: IU region contains a number of Sp1 sites and 1L region contains many homeo-specific sites and two octamers ATGCAAAT recognized by transcription factors Oct-1, Oct-2 and other POU domain containing proteins. Oct-1 sites may participate in oct-1 gene autoregulation. The role of U- and L-promoter fragments in oct-1 gene regulation was studied by transfection of luc-containing constructs in lymphoid (NS/0) and non-lymphoid cells. It was shown that L-promoter efficiency is much higher in lymphoid cells than in fibroblast cell line 10/1. U-promoter is active in both types of the cells. Upstream of IL-exon there is the inhibitory nucleotide sequence which elimination increases the expression level by many times. This silencing element is located between initiation starts of transcription and translation. The obtained data suggest that oct-1 gene contains at least two alternative promoters. It seems that U-promoter promises the constitutive synthesis of Oct-1 protein whereas L-promoter is tissue-specific and putatively is inducible and autoregulated. PMID- 16358732 TI - [Determination and analysis of the primary structure of NM.BstSEI operone from Bacillus stearothermophilus SE-589 which produces N.BstSEI site-specific nickase]. AB - Nucleotide sequence of Bacillus stearothermophilus SE-589 DNA fragment which includes an operone for site-specific NM-system with a gene for BstSEI nickase has been determined. Analysis of the regions adjacent to nickase gene has revealed two genes encoding DNA methyltransferases, which belong to different classes. Three genes which form system operone are separated with short open reading frames (ORFs). Analysis of these ORFs has shown that they encode polypeptides which are homologous to different parts of BstSEI nickase, NatB protein and arginase. A difference in GC-content of the beginning and ending regions of the cloned DNA fragment as well as presence of short ORFs similar to genes for known proteins may indicate that NM.BstSEI system operone has evolved by horizonthal DNA transfer. PMID- 16358733 TI - [Genetic variability and discriminating power of four microsatellite loci in Russian population]. AB - Allele and genotype frequencies of 4 STR loci (LPL, vWA, FES/FPS H F 13B), used in forensic medicine, were analyzed in Russian Siberian population. Genetic and molecular diversity of these polymorphic systems were characterized in comparison with US Caucasoid population. High discriminating power (PD = 0.99975) of the system of four studied STR loci was shown. Comparative analysis of genetic diversity in Russian population and Caucasoid US population revealed the significant differences between two populations and demonstrated that STR frequency data for US population should not be used for forensic expertise in Russia. The data obtained in the current investigation may be used as reference data for forensic medicine laboratories in Siberia. PMID- 16358734 TI - [Spatial organization of the chicken alpha-globin gene domain in cells of different]. AB - We have developed a technique for mapping the sites of DNA attachment to the nuclear matrix by hybridization of nuclear matrix DNA with an oligonucleotide array. The latter was made by immobilization of 60-mer oligonucleotides distributed within the area under study with a 2 Kb step on nylon filter. Using this approach we have analyzed the mode of interaction of a 100 Kb fragment of chicken chromosome 16 including the alpha-globin gene domain with the nuclear matrix. The 40 Kb DNA loop including all alpha-globin genes was detected in erythroid cells. One of the borders of this loop colocalized with the previously mapped MAR element and CTCF-dependent enhancer-blocking element. Also, a long transcribed area was found to be preferentially associated with the nuclear matrix. The spatial organization of the area under study in lymphoid cells was drastically different from this observed in erythroid cells. PMID- 16358735 TI - [Dynamic binding of the telomeric human protein TRF1 to intrachromosomal blocks (TTAGGG)n in living Chinese hamster cells depends on transcription]. AB - In some vertebrates tandem repeats (TTAGGG)n are located not only in telomeres, but also in intrachromosomal sites. In Chinese hamster cells such interstitial repeats which may be called "telomeric" heterochromatin (THC), representing up to 5% of genome. Earlier we have shown, that blocks of THC dynamically bind telomeric protein TRF1 in Chinese hamster cells. In this work question has been studied whether this interaction depends on a transcription. In cells with the normal transcription around 85% of initial fluorescence intensity of GFP-TRF1 is restored in 60 sec after the photobleaching. Treatment of the cells with transcription inhibitor actinomycin D (ActD) in the concentration completely inhibiting activity of DNA-dependent RNA polymerases I and II (RPI and RPII) leads to fast and practically full suppression of exchange GFP-TRF1 (10% of initial fluorescence is restored only) whereas an inhibitor of protein synthesis cycloheximide (CHD) has not effect. At the low ActD concentration, suppressing only RPI, efficiency of recovery of fluorescence was not changed. Since some fractions of heterochromatin in mammalian cells are actively transcribed, exchange of GFP-TRF 1 can be connected to transcription of THC which may be necessary for synthesis of small interfering RNA and self-maintenance of the heterochromatin, or with inhibition of expression of other genes effecting TRF1 stability. PMID- 16358736 TI - [Physico-chemical basis of RecA nucleo-protein filament formation on single]. AB - The analysis of RecA protein playing a central role in homologous recombination of E. coli with single-stranded DNAs of various structure and length on quantitative level is carried out for the first time. It was shown that weak additive interactions between protein monomers of filament and different structural elements of DNA provide DNA recognition. Orthophosphate and dNMPs (I50 = 12-20 mM) were shown to be the minimal inhibitors of RecA filamentation on d(pN)20. The lengthening of homooligonucleotides from d(pN)2 to d(pN)20 by one unit leads to monotonic increase in the affinity by a factor approximately 2 (factor f) due to weak additive contacts of RecA with every internucleoside phosphate group of DNA (f = 1.56) and specific interactions with each of T and C bases (f = 1.32). RecA filament does not practically interact with bases of d(pA)n, but contacts with internucleoside phosphate groups of the first turn (n < 10; f = 2.1) more effective than with additional turns of d(pA)n (n > 10; f = 1.3). The affinity of RecA protein for d(pN)n, containing typical and a number of different modified bases depends on a type of base, peculiarities of DNA structure and conformation of its sugar-phosphate backbone. The affinity is increased significantly if the bases contain exocyclic proton accepting groups. The possible reasons of preferable complexation of RecA with DNA of definite structure and length are analyzed. The mechanism of single-stranded DNA recognition by RecA and hypothetical mechanism of homological DNA strands exchange are proposed. PMID- 16358737 TI - [Positioning of mRNA 3' of the a site bound codon on the human 80S ribosome]. AB - Short mRNA analogues carrying a UUU triplet at the 5'-termini and a perfluorophenylazide group at either the N7 atom of the guanosine or the C5 atom of the uridine 3' of the triplet were applied to study positioning of mRNA 3' of the A site codon. Complexes of 80S ribosomes with the mRNA analogues were obtained in the presence of tRNAPhe that directed UUU codon to the P site and consequently provided placement of the nucleotide with cross-linker in positions +9 or +12 with respect to the first nucleotide of the P site bound codon. Both types mRNA analogues cross-linked to the 18S rRNA and 40S proteins under mild UV irradiation. Cross-linking patterns in the complexes where modified nucleotides of the mRNA analogues were in position +7 were analyzed for comparison (cross linking to the 18S rRNA in such complexes has been studied previously). The efficiency of cross-linking to the ribosomal components depended on the nature of the modified nucleotide in the mRNA analogue and its position on the ribosome, extent of cross-linking to the 18S rRNA being decreased drastically when the modified nucleotide was moved from position +7 to position +12. The nucleotides of 18S rRNA cross-linked to mRNA analogues were determined. Modified nucleotides in positions +9 and +12 cross-linked to the invariant dinucleotide A1824/A1825 and to variable A1823 in the 3'-minidomain of 18S rRNA as well as to protein S15. The same ribosomal components have been found earlier to cross-link to modified mRNA nucleotides in positions from +4 to +7. Besides, all mRNA analogues cross linked to the invariant nucleotide c1698 in the 3'-minidomain and to and the conserved region 605-620 closing helix 18 in the 5'-domain. PMID- 16358738 TI - [Investigation of folding/unfolding kinetics of apomyoglobin]. AB - Apomyoglobin kinetic and equilibrium unfolding and folding processes were studied at pH 6.2, 11 degrees C by stopped-flow tryptophan fluorescence. There are two distinct consecutive processes in apomyoglobin folding process, namely, the protein fast transition between the unfolded (U) and an intermediate (I) states (U <----> I) and slow transition between the intermediate and the native (N) states (I <----> N). Accumulation of the intermediate state was observed in the wide range of urea concentrations. The presence of the intermediate state was shown even beyond the middle transition on the unfolding limb. The dependence of observed folding/unfolding rates on urea concentration (chevron plot) was obtained. The shape of this dependence was compared with that of two-state proteins, folding from the U to N state. PMID- 16358739 TI - [Principal role of large polar residue clusters of RNA-binding proteins in the formation of complexes with RNA]. AB - Three-dimensional crystallographic data for a number of protein-RNA complexes were used to study the localization of protein binding regions at the protein surface. The set consists of ten non-homologous complexes, which include 20 independent binding sites. In most sites the contact interaction of polar residue side groups makes a significant contribution to protein binding regions. It was discovered that binding polar residues are localized essentially inside one largest cluster of polar residues at the protein surface. In fact, for fifteen regions 80.5% of polar binding residues were situated in such a large cluster. This result supports strongly a definitive role of the largest polar cluster in the recognition of the major groove of B-DNA by the protein transcription and regulatory factors, which has been recently disclosed by us. Therefore, at present one can suggest that the largest cluster of protein polar residues could play a key role in the formation of binding sites of proteins with DNA and RNA. PMID- 16358740 TI - [On prediction of folding nuclei in globular proteins]. AB - The approach described in this paper on the prediction of folding nuclei in globular proteins with known three dimensional structures is based on a search of the lowest saddle points through the barrier separating the unfolded state from the native structure on the free-energy landscape of protein chain. This search is performed by a dynamic programming method. Comparison of theoretical results with experimental data on the folding nuclei of two dozen of proteins shows that our model provides good phi value predictions for proteins whose structures have been determined by X-ray analysis, with a less limited success for proteins whose structures have been determined by NMR techniques only. Consideration of a full ensemble of transition states results in more successful prediction than consideration of only the transition states with the minimal free energy. In conclusion we have predicted the localization of folding nuclei for three dimensional protein structures for which kinetics of folding is studied now but the localization of folding nuclei is still unknown. PMID- 16358741 TI - [Development of a synthetic positive control which also detects plasmid contamination in diagnostic polymerase chain reaction]. AB - This paper describes a technique for the development of a positive control for use in a nested PCR to show that the PCR has worked correctly with both outer and inner primers designed for diagnostic amplification of 618 bp and 317 bp products respectively. This positive control produces a larger product than the diagnostic sample that can be discriminated on an agarose gel. This technique is advantageous over traditional cloning of the diagnostic PCR product itself by: 1) making it visually easy to detect plasmid contamination and thus, prevent false positives from the plasmid; 2) develop a positive control when the target organism is at a very low prevalence so initial detection is not relied on for cloning positive controls. This will ensure the PCR is working correctly prior to diagnostic sampling, reducing false negatives; or 3) for developing a PCR and determining the sensitivity prior to the use of diagnostic samples. The methods used to produce this nested positive control demonstrates how to use large oligonucleotide primers in PCR without non-specific binding occurring. PMID- 16358742 TI - [Fusion proteins encoded by orf 129L of ectromelia and orf A30L of smallpox viruses cross-react with neutralizing monoclonal antibodies]. AB - Open reading frame (orf) 129L of ectromelia (EV) and orf A30L of smallpox viruses (SPV) encoding fusion proteins were cloned and expressed in E. coli cells. The recombinant polypeptides (prA30L H pr129L) were purified from cell lysates by Ni NTA chromatography. Recombinant polypeptides were able to form trimers in buffered saline and they destroyed under treatment with SDS and 2 mercaptoethanol. Reactivity of prA30L, pr129L and orthopoxvirus proteins was analyzed by ELISA and Western blotting with panel of 22 monoclonal antibodies (MAbs) against orthopoxviruses (19 against EV, 2 MAbs against vaccinia virus and 1 Mabs against cowpox virus). This data allowed us to conclude that there are 12 EV-specific epitopes of pr129L and EV fusion proteins, ten orthopox-specific epitopes of EV, VV, CPV fusion proteins, from them 9 orthopox-specific epitopes of prA30L and SPV fusion proteins. Five Mabs, which cross-reacted with orthopox specific epitopes, were able to neutralize the VV on Vero cells and from them two MAbs has neutralizing activity against smallpox virus. Our findings demonstrate that 129L fusion protein have EV-specific epitopes, that EV 129L and SPV A30L fusion proteins have a several orthopox-specific epitopes to induce a neutralizing antibodies against human pathogenic orthopoxviruses. PMID- 16358743 TI - [Comparative study of variola virus and monkeypox virus interferon-gamma binding]. AB - DNA fragments containing genes for coding IFN-gamma-binding proteins (IFNgammaBPs) of variola virus (VARV) and monkeypox virus (MPXV) were obtained from viral genomes using PCR. Isolated genes coding desired proteins were expressed in the insect Sf21 cells using baculovirus expression system. Secreted recombinant IFNgammaBPs were isolated from culture medium of infected Sf21 cells through affinity chromatography procedure. SDS-PAAG and Western blot analysis of culture medium of infected insect cells and preparations of purified recombinant IFNgammaBPs indicated that recombinant viral proteins were dimerized even in the absence of ligand (hIFNgamma) unlike their cell (eucaryotic) analogs. Biological activity of the recombinant IFNgammaBPs were studied in the test of protective effect inhibition of hIFNgamma on L68 cells infected with murine encephalomyocarditis virus. It was shown that recombinant IFNgammaBPs had dose dependent IFNgamma-inhibiting activity. A possibility of the elaboration of new therapeutics for anti-hIFNgamma therapy on the base of IFNgammaBPs is discussed. PMID- 16358745 TI - [Gene expression regulation for amino acid biosynthesis and aminoacyl-tRNA synthetases in actinobacteria]. PMID- 16358744 TI - [Protease and reverse transcriptase genetic polymorphism in HIV type 1 subtype A variants predominating in cis countries]. AB - To define frequencies of drug resistance mutations among HIV-1 variants circulating within the territory of Russia, subtype A HIV-1 nucleotide sequences encoding protease and reverse transcriptase were analyzed. The analysis was carried out in 141 antiretroviral-naive individuals. Low frequency (less than 1%) of primary drug resistance mutations was shown. However, high frequencies of secondary mutations V77I in protease and A62V in RT (67% H 63%, respectively) linked to each other in most cases were observed. The HIV-1 isolates bearing both substitutions (MutV77I/A62V) were also characterized by the presence of several synonymous mutations, suggesting common origin for these viruses. HIV Biochip Hybridization microarray and/or Restriction fragment-length polymorphism analyses were performed to characterize gene pol polymorphism in additional 178 subtype A HIV-1 isolates. Among total 319 samples studied, Mutv77IA62V variant accounted for 56%, and was found to predominate in Russia in terms of both its geographical distribution and number of cases caused. Moreover, these viruses were prevalent in the regions known to have highest incidence of HIV-1 infection (Irkutsk, Samara, and Moscow regions). In addition, three other variants were found: viruses not containing the substitutions V77I or A62V, and variants bearing only one of them. Evolutional relationships between all four HIV-1 variants, as well as potential impact of the gene pol polymorphism on HIV-1 replicative fitness and drug resistance development are discussed. PMID- 16358746 TI - [Algorithm for large-scale finding of T box transcription regulation in bacteria]. PMID- 16358747 TI - [Ultrasonic and thermal inactivation of catalases from the bovine liver, the methylotrophic yeast Pichia pastoris, and the fungus Penicillium piceum]. AB - The kinetics of inactivation of catalases from bovine liver (CAT), the fungus Penicillium piceum (CAT1), and the methylotrophic yeast Pichia pastoris (CAT2) was studied in phosphate buffer (pH 5.5 or 7.4) at 45 and 50 degrees C or under the conditions of exposure to low-frequency ultrasound (LFUS; 27 kHz, 60 W/cm2). The processes were characterized by effective first-order rate constants (s(-1)): kin (total inactivation), k*in in (thermal inactivation), and k*in (us) (ultrasonic inactivation). The values of kin and k*in increased in the following order: CAT1 < CAT < CAT2. CD spectra of the enzyme solutions were recorded in the course of inactivation by high temperatures (45 and 50 degrees C) and LFUS, and the ratios of secondary structures were calculated. Processes of thermal and ultrasonic inactivation of catalases were associated with a decrease in the content of alpha helices and an increase in that of antiparallel beta structures and irregular regions (CAT1 < CAT < CAT2). We conclude that the enzymes exhibit the following rank order of resistance: CAT1 > CAT >CAT2. Judging from the characteristics of CAT1, it appears to be an optimum component for antioxidant enzyme complexes. PMID- 16358748 TI - [Trypsin-like proteinases and trypsin inhibitors in fruiting bodies of higher fungi]. AB - The activity of trypsin-like proteinases and trypsin inhibitors was measured in fruiting bodies of various species of basidial fungi (Basidiomycetes). Fruiting bodies of all fungi contained these enzymes, with the exceptions of polypore (Coriolus versicolor (Fr.) Karst) and hedgehog fungus (Hericium erinaceus (Fr.) Quel), belonging to the families Polyporaceae and Hericiaceae, respectively, in which the enzyme activities were barely detectable. The activity of trypsin-like proteinases was the highest in fruiting bodies of Boletaceae and Agaricaceae. Fruiting bodies of all fungi contained trypsin inhibitors. The highest activity of trypsin inhibitors was detected in basidiomycetes of the families Boletaceae, Agaricaceae, and Pleurotaceae, including Boletus castanus (Fr.) Karst, orange-cap boletus (Leccinum aurantiacum (Fr.) Sing), and brown-cap boletus (Leccinum melanum (Fr.) Karst). PMID- 16358749 TI - [Isolation of chitin-specific wheat oxidoreductases]. AB - Anionic peroxidase (IEP approximately 3.5) and oxalate oxidase (IEP approximately 7.0) were isolated from wheat seedlings using chitin. The strength of binding of enzymes with chitin depended on the degree of its acetylation and ionic strength of buffer. It was assumed that the acetyl groups of chitin are involved in sorption of enzymes on this biopolymer. The ability of anionic peroxidase and oxalate oxidase for sorption on chitin allows this biopolymer to be used for isolation of these proteins from plants. Cosorption of anionic peroxidase and oxalate oxidase on chitin suggests that these enzymes cooperate to ensure defensive response of wheat against chitin-containing pathogens. PMID- 16358750 TI - [Hydrogen peroxide content and catalase activity at inoculation with root tubercle bacteria of pea seedlings with the various nodulation ability]. AB - Hydrogen peroxide (H2O2) content and catalase activity were studied in pea (Pisum sativum L.) seedlings with normal (cultivar Marat) and disrupted (pea mutants) process of nodulation, which were inoculated with the nitrogen-fixing bacterium Rhizobium leguminosarum strain CIAM 1026. Differences in hydrogen peroxide content and catalase activity in pea seedlings with different ability for nodulation, which were inoculated with rhizobia, were found. It was assumed that H2O2 and catalase are involved in defensive and regulatory mechanisms in the host plant. PMID- 16358751 TI - [Bioconversion of beta-sitosterol and its complex esters by Rhodococcus actinobacteria]. AB - The ability of pure cultures of Rhodococcus actinobacteria from the Ural specialized collection of alkanotrophic microorganisms (World Federation for Culture Collections accession number 768; http://www.ecology.psu.ru/iegmcol) to convert beta-sitosterol (BSS) and its 3beta-acylated derivatives was studied. Rhodococcus strains with pronounced cholesterol oxidase activity, capable of converting BSS to stigmat-4-ene-3-one in the reaction of cooxidation with n hexadecane, were selected. The dependence of the activity of cholesterol oxidase of rhodococci on the length of the acyl group in BSS esters was studied. Conditions under which Rhodococcus cells convert BSS to 17beta-hydroxyandrost-4 ene-3-one (testosterone), commonly used in pharmacology, were determined. PMID- 16358752 TI - [Development of an oil-degrading biopreparation by activation of aboriginal hydrocarbon-oxidizing microflora]. AB - A method of activation of aboriginal hydrocarbon-oxidizing microorganisms for remediation of soil and water basins polluted with oil products was developed. The optimum composition of activating additives was found (g/l): mineral components, 10.0; oil, 5.0; and a synthetic detergent, 0.2. The resulting biopreparations increased the degree of purification by factors of 4-8 in soil and 18-24 in water when applied at a concentration of 10(7) cells/g(ml). PMID- 16358753 TI - [Inhibitory effects of phenolic ecotoxicants on photobacteria at various pH values]. AB - Kinetic characteristics of light emission by intact cells of the photobacteria Photobacterium phosphoreum and Vibrio harveyi at pH 5.5, 7.0, and 8.0 were studied as well as specific features of inhibitory effects of 2,4-di- and 2,4,5 triphenoxyacetic acids (2,4-D and 2,4,5-T), pentachlorophenol (PCP), and 2,6 dimethylphenol (2,6-DMP) at the same pH values. Nonstationarity of emission kinetics was observed at all the pH values studied. Exponential luminescence decay in a 60-sec range was observed at pH 5.5; a 5-min luminescence activation, at pH 7.0 and 8.0. The cell respiratory activity drops by over one order of magnitude at pH 5.5 compared with the activities at pH 7.0 and 8.0. The inhibitory effects of 2,4-D, 2,4,5-T, and PCP differ by one-two orders of magnitude depending on pH. The maximal cell sensitivity to these compounds appears at pH 5.5; the minimal, at pH 8.0. The effect of 2,6-DMP is independent of pH. As is demonstrated, it is hydrophobicity of the molecule and pK values of the toxicants that determine the inhibitory effect. Characteristic of the substrate-starved photobacterial cells are higher sensitivity to chlorophenolic compounds compared with the cells provided with high energy supply at all the pH values. PMID- 16358754 TI - [Cell aggregation in cultures of Micrococcus luteus studied by dynamic light scattering]. AB - Cell aggregation was studied using the method of dynamic light scattering in the course of growth of Micrococcus luteus cultures in a liquid medium. The method detects particles ranging in size from 0.5 to 1000 microm in samples containing no more than 10(5) cells/ml. When grown in liquid media, M. luteus forms aggregates; during the lag phase, 80% of the cells are found in aggregates of 10 to 1000 microm, only minor amounts being represented by single cells. With the onset of exponential growth, the aggregates were decomposed, and single cells became prevalent in the culture liquid. This observation confirms that the aggregation of the cells during the lag phase is prerequisite to the initiation of bacterial growth. The method may be used in biotechnology for monitoring the state of bacterial cultures. PMID- 16358755 TI - The changes of catechins during the fermentation of green tea. AB - The dynamics of tea catechins and organic acids in fermented fluid and yeast cells were studied. The concentration of eight kinds of catechins solution decreased by from 29.6% to 47.6%, respectively, some catechins were absorbed and accumulated by yeast cells, but the amount in the cells was very low during the fermentation process. The investigation of catechins resolved in four citrate buffers with a pH range of 2.6-5.6 for 18 h showed that most catechins were stable in buffer solutions of pH 4.6 and 5.6, and significant losses took place in solutions of pH 2.6 and 3.6. However, most catechins were released and recovered by adjusting the pH value to 5.6, which suggested that catechins in extremely acidic buffer solutions might reversibly combine each other or with other compounds. PMID- 16358756 TI - [Genetic differentiation of the sherry yeasts Saccharomyces cerevisiae]. AB - Molecular genetic study of the yeast Saccharomyces cerevisiae isolated at various stages of sherry making (young wine, solera, and criadera) in various winemaking regions of Spain demonstrated that sherry yeasts diverged from the primary winemaking yeasts according to several physiological and molecular markers. All sherry strains independently of the place and time of their isolation carry a 24 bp deletion in the ITS 1 region of ribosomal DNA, whereas the yeasts of the primary winemaking lack this deletion. Molecular karyotypes of the sherry yeast from different populations were found very similar. PMID- 16358757 TI - Decolorization of alcohol distillery wastewater by thermotolerant white-rot fungi. AB - To detect thermotolerant fungus strain for decolorization of alcohol distillery wastewater (WAD), 38 fungus strains were studied. Ability of ligninolytic enzyme production was examined at 35 and 43 degrees C on agar media containing 2,2' azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) and MnCl2. At 43 degrees C, four of Pycnoporus coccineus strains showed their higher potential for WAD decolorization both on agar media and in liquid media. Immobilized mycelia on polyurethane foam removed total phenol about threefold higher than free mycelia did in shaking condition at 43 degrees C. Moreover, color removed by immobilized mycelia nearly 50% higher than free mycelia did. PMID- 16358758 TI - [Enzyme immunoassay of ampicillin in milk]. AB - An indirect immunoassay for quantitative determination of ampicillin (range, 10 1000 ng/ml) in buffer or milk has been developed. Polyclonal antibodies were obtained against ampicillin conjugated with bovine serum albumin; the conjugate was synthesized by direct condensation using carbodiimide. The antibodies were specific for ampicillin and exhibited low cross-reactivity to other penicillins (azlocillin, 17%; penicillin G, 10%; piperacillin, 5%; and carbenicillin, 4%). Matrix effects were minimized by combining the use of a casein-supplemented buffer (content of casein, 1%) with sample dilution. The threshold of ampicillin detection in milk (diluted tenfold) was equal to 5.0 ng/ml (which corresponded to 50 ng/ml of the original sample). PMID- 16358759 TI - [Phenol conjugation with peptides and final transformations of conjugates in English ryegrass seedlings]. AB - [ 1-14C] Phenol transformation in English ryegrass (Lolium perenne L.) sterile seedlings was studied. The compound studied was assimilated by a plant through leaves as vapor. Phenol was bound with ryegrass low-molecular-weight peptides producing phenol-peptide conjugates. Conjugation with endogenous peptides is one of the main pathways of phenol detoxication in ryegrass. Nearly three fifths of phenol assimilated is conjugated with low-molecular-weight peptides. After removal of the plant from the labeled phenol-containing atmosphere, the content of conjugation products gradually decreased, followed by excretion of labeled carbon dioxide. This fact indicates that the conjugates are destructed and the carbon atoms of their radioactive component are oxidized to carbon dioxide. Almost one third of assimilated phenol is transformed via the oxidation pathway, but a small part of it is irreversibly bound with biopolymer molecules. PMID- 16358760 TI - [Response of winter wheat to cold: production of phenolic compounds and L phenylalanine ammonia-lyase activity]. AB - The formation of soluble and polymeric (lignin) phenolic compounds, activity of L phenylalanine ammonia lyase (PAL, EC 4.3.1.5), and content of free L phenylalanine during cold hardening of winter wheat plants (Triticum aestivum L.) were studied. Cold treatment increased accumulation of soluble phenolic compounds in leaves while not affecting the content of lignin. The opposite was observed in tillering nodes. The activity of PAL was lower than in control plants in both tissues, and the content of free L-phenylalanine in tissues increased. PMID- 16358761 TI - [Oxidase-peroxidase method for determination of ethanol in fermented musts and wine products]. AB - A new alcohol oxidase-peroxidase method of determination of ethanol content in fermented musts and wine products is described and compared to conventional methods routinely used in winemaking. The sensitivity, accuracy, and reliability of this method were determined. The results of ethanol determination in fermented musts and wines correlated well with the data obtained by refractometry (correlation coefficient R = 0.9595, p < 0.0001) and densitometry (correlation coefficient R = 0.9384, p < 0.0001). This method is less time- and labor consuming and allows simultaneous testing a series of wine samples. PMID- 16358762 TI - [Antioxidant properties of essential oils: autoxidation of essential oils from laurel and fennel and effects of mixing with essential oil from coriander]. AB - Changes in the composition of essential oils from the seeds of laurel (Laurus nobilis L.) and fennel (Foeniculum vulgare Mill., var. dulce Thelling) and their mixture with essential oil from coriander were studied by capillary gas-liquid chromatography during storage in the dark and in light. Under these conditions, essential oil of laurel retained its composition for 12 months. Essential oil of fennel was rapidly oxidized in light. However, the rate of its oxidation in the dark was lower. The major component of essential oil of fennel, transanethol, had a lower antioxidant activity than essential oil of coriander. The mixture of essential oils from laurel and coriander possessed antioxidant properties and strongly inhibited the oxidation of components of the fennel oil. PMID- 16358763 TI - [The All-Russia Symposium on Biotechnology of Microbes]. PMID- 16358764 TI - [International Scientific and Practical Conference: Prospects and Problems in the Development of Industrial Biotechnology within the Common Economic Space of the Commonwealth States. Minsk-Naroch', May 2005]. PMID- 16358765 TI - [Problem of terminology in characteristics of spermatozoa of metazoa]. AB - Difficulties in characterization of spermatozoa of the Metazoa are now related to insufficiency of the traditional terms "primitive spermatozoon", "modified spermatozoon", and "aberrant spermatozoon" introduced by G. Retzius and A. Franzen for description of intermediate forms discovered in the 20th century. In this respect, some authors propose to reject the Retzius' and Franzen's terms and turn to the terminological system of D. Rouse and B. Jamieson, the terms of which ("ectaquaspermatosoa", "entaquaspermatozoa", and "introspermatozoa") determine specific features of insemination, but do not reflect the structure of spermatozoa. The assertion of "helplessness" of the traditional structural Retzius' and Franzen's terms is unfounded. Their preservation is desirable if one wishes to preserve the comparative-morphological approach to the characterization of male gametes of the Metazoa. The terms "primitive spermatozoon", "modified spermatozoon", and "aberrant spermatozoon" will become more universal if the method of their combined utilization is applied. PMID- 16358766 TI - [Regulation of development of wing venation in Drosophila melanogaster by a network of signalling pathways]. AB - The paradigm of contemporary phenogenetics consists in that ontogenetic development of animals is regulated by a set of cell signalling pathways. The main signalling pathways controlling the wing development are Hedgehog, Notch, Wingless, Epidermal growth factor receptor, and Decapentaplegic. Expression of many genes in the imaginal disc is specific for both cells competent for transformation into veins and adjacent cells forming wing cells. Individual genetic programs control differentiation of each longitudinal vein and determination of its position on the wing. The gene net plays an important role in wing venation of D. melanogaster. Mutations causing some ectopic expressions of transcriptional factors and the activities of signalling cascades are, possibly, involved in phylogenetic transformation of the insect wing. PMID- 16358767 TI - [Role of telomerase in reactivation of macrophage nuclei in heterokaryons]. AB - It was shown that the duration of stay of macrophages in the peritoneal cavity of mice and method of their isolation did not affect markedly their capacity for resumption of DNA synthesis in heterokaryons. This means that mouse macrophage undergo such changes during differentiation that reactivation of DNA synthesis in their nuclei is only possible after interaction of telomeres with telomerase, since it was already shown that telomerase was involved in reactivation of DNA synthesis in the macrophage nuclei. The results of experiments did not reveal differences in the length of telomeres in mouse macrophages and other somatic cells. This could depend on the significant length of mouse telomeres and, as a result, their shortening, sufficient for the inhibition of proliferation, is beyond the limits of sensitivity of the current methods. It is also possible that changes in DNA properties in the macrophages occurring during their differentiation depend on changes in the conformation of the telomere complex in these cells. Testing of this suggestion is relevant with respect to recent data that cell hybridization, specifically in the form of heterokaryons, may be essential in realization of the therapeutic effect caused by the introduction of cells during cell therapy. PMID- 16358768 TI - [Catecholamines in regulation of development of GnRH neurons of rat fetuses]. AB - The contents of dopamine, serotonin, and noradrenaline in rat fetuses developing under the conditions of their deficiency induced by administration of alpha methyl-para-tyrosine to females during 11th to 16th or 20th day of pregnancy and in fetuses, whose mothers were given saline at the same time, were determined using HPLC with subsequent electrochemical detection. Administration of alpha methyl-para-tyrosine led to decreased levels of dopamine and noradrenaline in the areas of migration of GnRH-neurons in fetuses on days 17 and 21 of prenatal development. The concentration of serotonin remained unchanged, except in the head nasal area in males on day 21. The areas of interaction between the brain catecholaminergic systems and migrating and differentiating GnRH-neurons were determined by double immunohistochemical labeling. Close topographical location of GnRH-immunoreactive neurons and tyrosine hydroxylase-immunoreactive in the area of nucleus accumbens on days 17 and 20, as well as in the median eminence on day 20. The GnRH concentration in the caudal areas of migration of GnRH-neurons under the normal conditions and in the case of catecholamine deficiency was determined using radioimmunoassay. After administration of alpha-methyl-para tyrosine the GnRH concentration in the anterior hypothalamus decreased in females. The data obtained suggest the involvement of catecholamines in the regulation of development of GnRH-Neurons during prenatal development. In addition, the adequacy and efficiency of the used model of catecholamine deficiency for studying the development of such neurons was confirmed. PMID- 16358769 TI - [Changes in composition of acid soluble proteins and DNA in chromatin of rat liver and brain bound and not bound to nuclear envelope as a function of age and under the influence of antioxidant ionol]. AB - In two-day rat pups, the histone H1 content in the brain chromatin was higher than in the liver chromatin, as compared to histone of the nucleosome core. The H1 content in the brain chromatin decreased with the age, while in the liver chromatin it increased. At the same time, in the adult brain chromatin bound to the nuclear envelope, a high level of H1 characteristic of chromatin of the newborn rats was preserved, while in a similar chromatin of the adult liver, the H1 content increased, but still remained less than in the chromatin not bound to the nuclear envelope. In both organs, the composition and quantitation of H1 subfractions were different in chromatins bound and not bound to the nuclear envelope. The chromatin from the liver and brain bound to the nuclear envelope differed also in the composition and quantitation of minor acid soluble proteins. In the presence of the antioxidant ionol, the 5-methylcytosine content in DNA of chromatin of the rat liver bound to the nuclear envelope increased while in the chromatin not bound to the nuclear envelope, it remained unchanged. Thus the chromatins bound and not bound to the nuclear envelope differ in the composition and mount of acid soluble proteins, including histone H1, the contents of these proteins in bound and not bound chromatin are different and change with the age in different ways. The antioxidant ionol affects differently the methylation of bound and not bound chromatin. PMID- 16358772 TI - [Outstanding embryologist of the 20th century Boris I. Balinsky. Postscript to the paper of Vladimir P. Korzh "Introduction of boris balinsky to embryology"]. PMID- 16358773 TI - [Hydration enthalpy characteristics of amino acids in solutions]. AB - The enthalpies of solvation of 17 amino acids were evaluated by using the sublimation enthalpies of amino acids and the standard enthalpies of their solution in water. An equation was derived, which relates the volume-specific enthalpy of sublimation (deltaH(subl)/V(w)) to the sum of the common bond lengths in molecules (sigman(i)l(i)) of substances examined. The results obtained are interpreted in terms of the effect of hydrophobic and hydrophilic side chain on the interactions between the zwitterions of amino acids and water molecules. PMID- 16358774 TI - [Molecular dynamics of oligopeptides 6. A comparative study of poincare cross section of monopeptide frames in media with different hydrophobicity]. AB - A comparative study of the molecular dynamics of natural amino acid residues and their closest homologues and isomers was carried out. Molecular dynamics protocols not interfering with the principle of equidistribution of energy with respect to degrees of freedom were used. Poincare cross-sections, auto- and cross correlation of complex exponential curves as a function of dihedrons were considered. The classification of dynamic properties of conformational degrees of freedom in the series of amino acid residues was carried out. PMID- 16358775 TI - [Investigation of the amino acid sequences of open reading frames of the Mycobacterium tuberculosis complete genome with the use of the protein family pattern bank Prof_Pat]. AB - The new version of protein family patterns bank Prof_Pat 1.16 was used to investigate 3924 amino acid sequences of open reading frames of Mycobacterium tuberculosis strain H37Rv. The similarity to proteins with the known function was confirmed for 2772 amino acid sequences. Possible functions for 44 thus far unidentified sequences of open reading frames were determined. PMID- 16358776 TI - [The volume and structure of the chymotrypsin active site]. AB - It was found that the chymotrypsin active site is located in the largest cleft on the enzyme surface approximated by a sphere with a radius of 20 angstroms. The active site cleft volume is about 2 nm3, as computed by the Monte-Carlo method. The size and shape of the active site cleft-- the intersection of two unequal spheres--are sufficient for large (about 1 nm3) fragments of substrate molecules to enter the active site. The active site bottom and the adjacent narrow section are about 600 angstroms3 in volume and may serve as a combustion chamber of a water-substrate mixture during the operation of the enzyme machinery. Intrinsic water molecules inside the combustion chamber can take part in heat exchange during different steps of the enzymatic process. PMID- 16358777 TI - [A comparison of X-ray diffraction analysis and nuclear magnetic resonance from the data on the identification of alpha-helices and beta-strands in the same proteins]. AB - The methods of X-ray diffraction analysis and nuclear magnetic resonance spectroscopy were compared using the data on the identification of alpha-helices and beta-strands of the same proteins. The goal of the study was to determine whether these identifications can be considered as equivalent in the structural classification of proteins and in the solution of other problems. The identifications obtained by the method of Kabsch and Sander for 56% specially chosen water-soluble proteins were chosen. It was found that the identification of alpha-helices and beta-strands are almost equivalent if used for the structural classification of the proteins. In the analysis of the conformational properties of amino acid residues or their combinations, it is reasonable to use the identifications of alpha-helices and beta-strands, obtained only from the data of X-ray diffraction analysis. An additional outcome of the study is a unique collection of pairs of protein structures obtained by the methods of X-ray diffraction analysis and nuclear magnetic resonance spectroscopy for the same proteins. PMID- 16358778 TI - [A study of protein structure changes during hydration by diffuse X-ray scattering. I. The intensity and the shape of "10-angstrom" maximum]. AB - The angle dependencies of diffuse x-ray scattering intensities were studied in a wide range of angles from 3 to 80 degrees for water-soluble and membrane proteins with a different structural organization: alpha-helical protein myoglobin, alpha helical protein serum albumen, alpha + beta protein lysozyme, and transmembrane proteins of photosynthetic reaction centers (RC) from purple bacteria Rhodobacter sphaeroides, and Blastochlorii (Rhodopseudomonas) viridis containing cytocrome c, situated out side the membrane, and for H and L+M subunits of membrane protein of reaction center from Rb. sphaeroides for various hydration degrees. The hydration/dehydration process was studied for water-soluble proteins (within hydration range from h = 0.05 to h = 1). The hydration/dehydration process appears to be reversible. All water-soluble proteins show a 10 angstroms peak, and proteins of reaction center do not show this peak. A quantitative comparable study of the behaviour for of the 10 angstroms peak different proteins the degree of lysozyme hydration increases from h = 0.05 to h = 0.45, the protein structure slightly changes (most probably the motifoffolding), the structure of myoglobin in solution is slightly different from the structure in crystal. By taking into account the changes in the shape and intensity of the 10 angstroms peak only, it is impossible to make the conclusion about structural changes in other proteins studied. A correlation between the structural changes observed and dynamic and functional properties of proteins is discussed. PMID- 16358779 TI - [Photochemical revelation of the stacking aggregation of thymine chromophores in water solutions of polythymidylic acid]. AB - The structure heterogeneity of water solutions of polyribothymidylic acid at T(room) was studied from changes caused in their absorption spectra by the photodimerization reaction. Three fractions of thymine chromophores were revealed from the differential absorption spectra: (a) the main fraction consisting of weakly interacting (isolated chromophores) chromophores with the absorption spectrum maximum at approximately 270 nm; (b) pair chromophores of the first type with the absorption spectrum maxima at 260 and 290 nm (exciton splitting 4000 cm( 1)); and (c) pair chromophores of the second type with the absorption spectrum maxima at 250 and 280 nm (exciton splitting 4300 cm(-1)). The revealed aggregates have a relatively high photochemical activity in the photodimerization reaction in comparison with the isolated chromophores. They contribute little to the total absorption spectrum of solutions but make a great contribution to its changes at the initial stages of the UV irradiation of solutions. PMID- 16358780 TI - [The visualization of amplicons after polymerase chain reaction]. AB - Linear DNA molecules amplified by the polymerase chain reaction were visualized by atomic force microscopy. The measured contour length of the PCR product of 1414 bp sequence was 435 +/- 15 nm. Considering that the calculated value of the distance between the nucleotides along the duplex axis is 0.31 nm, it was assumed that linear DNA molecules on the surface of mica, which serve as a support in the atomic force microscopy method, are in the A form. The influence of surface properties of the mica and the sample drying procedure on the conformation of adsorbed DNA molecules is discussed. Possible reasons for the Gaussian distribution of the contour length of the synthesized amplicon are considered. PMID- 16358781 TI - [DNA-binding and antiviral activities of bis-netropsins]. AB - The DNA-binding and antiviral activitus of bis-netropsins in which two monomers are attached covalently via three glycin residue were studied. These compounds have the same C-end groups but contain clusters with different numbers of lysine residues at the N-end of the molecule. In the homologous series of these compounds, bis-neropsins containing 15 and 31 branched lysine residues at the N end of the molecule appear to be the most effective inhibitors of reproduction of the simplex herpes virus of type I in the Vero cell culture, including the virus versions resistant to aciclovir, ganciclovir, and other medicinal preparations. It was shown that the cytotoxicity of all the compounds studied is much lower than that of netropsin. The antiviral activity of the compounds correlates with their ability to selectively interact with the expanded clusters of the AT-pairs of DNA bases in the form of a monomer or a dimer, stabilized by interaction between the C-end halves of two bis-netropsin molecules bound at the neighboring overlapping binding sites on the DNA. The possible sites of their binding are the expanded clusters of AT-pairs at the origin of replication of OriS and OriL of the herpes virus. PMID- 16358782 TI - [The thermodynamic stability of nanoconstructions based on the double-stranded DNA molecules]. AB - The temperature stability of nanoconstructions formed by double-stranded DNA molecules fixed in the structure of their liquid crystalline dispersions and cross-linked by nanobridges was determined. It was shown that the heating of nanoconstructions is accompanied by a decrease in the amplitude of the negative bands in the CD spectrum both at lambda approximately 310 and lambda approximately 505 nm. Temperature "melting curves" were derived and characterzed by T(M) values. The T(M) values at lambda approximately 310 and lambda approximately 505 nm coincided with each other but differed from the T(M) value characteristic of the DNA cholesteric liquid crystalline dispersion. PMID- 16358783 TI - [The dynamic heterogeneity of the phospholipid bilayer and diffusion of molecules at the interface]. AB - The method of steered molecular dynamics was developed to evaluate the kinetic parameters of the penetration of molecules through the interface. Heterogeneous microviscosity of a membrane was calculated by the example of a hydrated 1 palmitoyl-2-oleyl-sn-glycero-3-phosphatidylcholine bilayer. The influence of the chemical nature of the penetrant molecule on its penetration through the membrane was studied. PMID- 16358784 TI - [A mathematical model of conformational changes in membrane protein part of F0F1 ATP synthase]. AB - The transformation of the electrochemical membrane proton gradient into the energy of chemical bond in adenosinetriphosphate is one of the most important biological processes occurring in the living cell. The main enzyme that directly catalyzes the synthesis of ATP from ADP and Pi in aerobic conditions is F0F1-ATP synthase. In the present study, conformational changes in membrane protein part of ATP synthase during its catalytic cycle were described in terms of dynamic equations of solid state rotation. It was shown that this process should be considered in the framework of classical mechanics. The time dependences of the angle rotation for the protein complex rotates around its central axis were oobtained. Possible types of rotation were analyzed. It was proved that, considering the modern level of knowledge and understanding of ATP synthase structure and function are taken into account, the minimal period of rotor turnover cannot be less than 10(-9) s. With regard for viscous friction and elastic tension in the central axis, the calculated time of whole turnover varies from 10(-6) to 10(-3) and depends on the set of parameters used. These results indicate the essential contribution of friction and elastic tension to the rotation dynamics of the rotor. It is suggested that the proposed model can be used as a part of the entire algorithm for computer simulation of ATP synthase. PMID- 16358785 TI - [The calmodulin inhibitor R24571 induces a short-term Ca2+ entry and a pulse-like secretion of ATP in Ehrlich ascites tumor cells]. AB - The properties of the Ca2+ channel induced by a calmodulin inhibitor in Ehrlich ascites tumor cells were investigated using fluorescent indicators Indo-1 and chlortetracycline. The inhibitor of calmodulin calmidazolium (R24571) in concentrations of 1-2 microM induces a short-term Ca2+ entry and a pulse-like ATP secretion. Repeated addition of R24571 also causes a transient Ca2+ signal. Ca2+ channels induced by R24571 are permeable for Mn2+. Ca2+ entry does not depend on endoplasmic reticulum depletion by thapsigargin, ATP, or ionomycin and is suppressed by nordihydroguaretic acid (EC50 = 6.7 microM), quercetin (EC50 = 1.5 microM), dihydroquercetin (EC50 = 17 microM), arachidonic acid (AA) (EC50 = 8.6 microM), and suramin (EC50 = 0.25 +/- 0.05 MM), and weakly depends on temperature in the range of 18 - 37 degrees C. The apparent activation constant for R24571 and the Hill coefficient are 2.5 +/- 0.2 and 4 +/- 0.3 microM, respectively. The products of arachidonic acid oxidation are neither activators nor inhibitors of these channels. The inhibitory effect of nordihydroguaretic acid is indirect and is conceivably caused by the accumulation of arachidonic acid due to suppression of its lipoxygenase-catalyzed oxidation at phospholipase A2 activation. The maximal level of about 1.3 microM in the dependence of Ca2+ signal amplitude on R24571 concentration points to possible inhibition of the channel by increased Ca2+ concentration in the cytosol. The weak dependence on temperature implies that the channel is highly permeable, the chain of enzymic processes is not involved in Ca2+ entry activation, and the mutual compensation of processes with opposite contributions is possible. Using chlortetracycline fluorescence, we have shown in model experiments on calmodulin solution that Ca2+ induces cooperatively a conformational transition of calmodulin with the exposure of a hydrophobic chlortetracycline-Ca(2+)-binding site. The interaction of R24571 with the CaM Ca2+ complex results in quenching of fluorescence to its level in water, which is interpreted as the elimination of the availability of calmodulin hydrophobic site for chlortetracycline-Ca+. Nordihydroguaretic acid, quercetin, and dihydroquercetin, but not suramin, also interact with calmodulin, but this does not result in the complete closing of its hydrophobic site. It is supposed that the activation of the Ca2+ channel occurs owing to the activation of calmodulin dependent phospholipase A2 by R24571, which leads to the formation of a low molecular short-lived secondary messenger, or because of the interaction of R24571 with calmodulin, which directly inhibits the channel. The termination of Ca2+ entry is probably due to the inhibition of phospholipase A2 and/or of the channel at increased concentrations of arachidonic acid and Ca2+. PMID- 16358786 TI - [Photochemical activity of photosystem II and hydrogen photoproduction in sulfur deprived Chlamydomonas reinhardtii mutants D1-R323D and D1-R323L]. AB - The role of photosystem II in hydrogen photoproduction by Chlamydomonas reinhardtii cells was studied in mutants with modified D1-protein. In D1-R323D and D1-R323L mutants, the replacement of arginine by aspartate or leucine, respectively, resulted in the disruption of electron transport at the donor side of photosystem II. The rate of oxygen evolution in D1-R323D decreased twice as compared to the pseudo-wild type (pWT), and in D1-R323L no oxygen evolution was detected. The latter mutant was not capable of photoautotrophical growth. The dynamics of changes in oxygen content, the reduction of photosystem II active reaction centers (deltaF/F(1)m), and hydrogen production rate in pWT were found to be similar to the wild type if cultivated under sulfur deprivation in a closed bioreactor. The observed gradual decrease in the deltaF/F(1)m value turned to a sharp drop almost to zero followed by a partial recovery during which the production of hydrogen set in. The transition to the anaerobic phase in D1-R323D cultured in a sulfur-deprived medium occurred earlier than it happened in pWt under the same conditions. However, the partial recovery of photosystem II activity and hydrogen production started at a later time, and the rate of hydrogen production was low. The D1-R323L mutant incapable of oxygen evolution entered the rapidly anaerobiosis but produced no hydrogen. The kinetics of photoinduced redox transitions in P700 was similar in all investigated strains and was not affected by diuron addition. This implies that the mutants had a pool of reducers, which could donate electrons through the quinone pool or cytochrome to photosystem I. However, in D1-R323L mutant lacking the active photosystem II, this condition was not sufficient to support hydrogenase activity. PMID- 16358787 TI - [The isotope effect in the glycine dehydrogenase reaction is the cause of the intramolecular isotope inhomogeneity of glucose carbon of starch synthesized during photorespiration]. AB - The isotope distribution of glucose-6-phosphate in the main pathways of its biosynthesis (in the processes of CO2 assimilation and photorespiration in the Calvin cycle and during resynthesis from the degradation products of lipids and proteins) was analyzed. For reconstructing the isotope distribution of glucoso-6 phosphate synthesized in the Calvin cycle during photorespiration, the functioning of the cycle with regard to its coupling with the glycolate chain, which together constitute the photorespiration chain, was considered. In the glycine dehydrogenase reaction of the glycolate cycle, there arises an isotope effect, which determines the distribution of isotopes in the glucose-6-phosphate and other photorespiration products. The isotope effect of the glycine dehydrogenase reaction increases at the expense of the exhaustion of glucose resources feeding the photorespiration chain. As a result, atoms C-3 and C-4 of glucose become enriched with the heavy isotope, and subsequent mixing of atoms and the specificity of interactions in the photorespiration chain lead to an isotope weighting of the other atoms and an uneven distribution of carbon isotopes in glucose-6-phosphate and other photorespiration products. A comparison of the glucose-6-phosphate isotope patterns in different pathways of the synthesis with the experimental data on the distribution of carbon isotopes in starch glucose of storing plant organs led to the conclusion that the starch resources are predominantly formed at the expense of glucose-6-phosphate of photorespiration. This is consistent with the earlier observed enhancement of photorespiration at the stage of plant maturation. PMID- 16358788 TI - [Modeling of hysteresis in pH pattern formation along the cell membrane of algae Chara corallina]. AB - It is known that illumination of the algae Chara corallina results in the formation along the membrane of regions with inhomogeneous distribution of pH. It was shown that, in a particular range of illumination intensities, two states with different pH distribution are realized at one and the same value of light intensity: an entirely homogeneous state and completely formed structures (pattern). The transition from the homogeneous state to the pattern formation takes place at one value of light intensity, and the back transition, at another light intensity, i.e., the hysteresis is observed. This phenomenon was studied by mathematical modeling. The mechanism of hysteresis is discussed. PMID- 16358789 TI - [Effects of the thiol reagent dithiothreitol on the oxidation-reduction potential and the growth of Escherichia coli in anaerobic conditions at different pH]. AB - Changes in the oxidation-reduction potential of medium during the growth of Escherichia coli bacteria under anaerobic conditions in the pH range 5.5-7.5 were studied. It was shown that dithiothreitol, a reducer of thiol groups, at a concentration of 3 mM decreased the initial value of oxidation-reduction potential to negative values of -60 to -220 mV, increased the duration of the lag growth phase, decreased the growth rate, and lowered the acidification of the medium and accumulation of potassium ions independently of pH. The value of the potential in the presence of 3 mM dithiothreitol decreased by 15-20 mV both at alkaline and acidic pH. Dithiothreitol at a concentration of 10 mM stopped molecular hydrogen production at pH 5.5 only. The effects of dithiothreitol on the oxidation-reduction potential and bacterial growth were probably associated with changes in the state of thiol groups in membrane proteins, which affects the generation of membrane potential, proton secretion from cells, K+ accumulation in bacteria, and the activity of membrane-associated enzymes. PMID- 16358790 TI - [Chemiluminescence in a stimulated polymorphonuclear leukocytes--luminol system: suppression by thiols]. AB - The effect of some scavengers of thiol nature, which eliminate all reactive oxygen species and oxidants with reactive chlorine, on the luminol-enhanced chemiluminescence of polymorphonuclear leukocytes was studied. The use of two scavengers of this type (penetrating and not penetrating into the cell) made it possible to separate the luminescence of cell structures from the luminescence generated by oxidants in the surrounding medium. It was found that about a half of luminol luminescence is due to its oxidation in the medium surrounding the cell, and it is completely inhibited by the nonpenetrating reduced glutathione. The cell itself is a source of a considerable portion of luminescence, and this luminescence is quenched by penetrating sulfhydryl compounds such as dithiothreitol and N-acethyl cysteine. Reduced glutathione, which penetrates into cells and whose action is due only to the sulfhydryl group, is recommended as a candidate for the selective neutralization of extracellular oxidants. PMID- 16358791 TI - [A mathematical model of delayed luminescence oscillations of higher plants and analysis of the reaction rates calculated by the model]. AB - The oscillatory regime of delayed millisecond luminescence was obtained by the model developed earlier. We compared the rates of changes in the concentrations of some of the metabolites calculated by the model with the rates calculated by other known models. The results of calculations for the changes in metabolite concentrations after switching off the light were compared with experimental data. PMID- 16358792 TI - [Fluorescence parameters of chlorophyll in leaves of caules plants in different environmental conditions]. AB - The functional state of medicinal plants of Convallaria majalis L., Vaccinium vitis-idaeae L., Arctostaphylos uva-ursi L. in connection with heavy metal accumulation in their leaves under man impact was studied by the pulse-amplitude modulation (PAM) fluorometric method. The relative yield of variable fluorescence (F(v)/F(m)), induction of fluorescence of chlorophyll, and fluorescence quenching processes in leaves at different distances from the local Kirov-Sovetsk, Kirov Omutninsk road in Kirov region were analyzed. Changes in biophysical characteristics with the increasing content of heavy metals in leaves were demonstrated. The most informative characteristic is F(v)/F(m). Its value correlates with the activity of the photosynthetic apparatus and reflects the potential effeciency of photosynthesis. The better are the environmental conditions of plant growth, the higher is the F(v)/F(m) ratio and the lower is its average statistical deviation. Fluorescence induction curves do not always vary in shape under our ecological conditions, indicating relatively favorable conditions at places of plant growth investigated. The rate of the environmental pollution in the investigated region is not critical, since the content of heavy metal in leaves does not change considerably with the distance from the road. PMID- 16358793 TI - [Lanthanides induce neurotransmitter release from the vesicular pool in rat brain synaptosomes]. AB - The influence of lanthanoids on exocytosis was investigated. It was shown that gadolinium increases the spontaneous release of the glutamate nonmetabolizing analogue [3H]D-aspartate. It was established using the fluorescent dye acridine orange that gadolinium and lanthanum induce exocytosis. The effect was dose dependent and was maximum at 300 microM Gd3+. The exocytosis induced by gadolinium was calcium-independent. It is suggested that lanthanides induce a vesicular release of neurotransmitters by the mechanisms common for all polyvalent cations. PMID- 16358794 TI - [Mathematical modeling of the mechanisms of locomotory pattern formation under epidural spinal cord stimulation with consideration of peripheral feedback]. AB - The mechanisms of nervous regulation of locomotory activity of the spinal cord and participation of afferent peripheral feedback from lower limb muscles in the formation of locomotory patterns were investigated. The set of electromyograms of lower-limb muscle groups recorded in experiments on mesencephalic cats with application of electric epidural stimulations of lumbar segments of the spinal cord is described by a nonlinear dynamic model constructed on the basis of the Van-der-Pol equation with the compelling member. The conditions of occurrence of the regime of self-oscillations were investigated depending on the parameters of external influence. A modified equation was proposed, which takes into account the role of the afferent feedback and delay between the beginning of stimulation and muscle reaction. The conformity of the mathematical model with experimental data was shown, which makes possible its use both for the description of the mechanism of locomotory pattern formation under epidural spinal cord stimulation and the choice of optimum stimulation conditions. PMID- 16358795 TI - [Effect of toluene on stability of chromatin of white rat hippocampus and olfactory bulb in vivo. Microcalorimetric study]. AB - The denaturation heat parameters of hippocampus and olfactory bulb nodulus tissues were determined. The total denaturation heat calculated from the areas of endotherms I-IX onto which the dependence deltaH = f(T) is factorized equals to 13.03 +/- 1.3 J/g for bulb nodules and 9.91 J/g for the hippocampus. It was shown that chromatin in the composition of tissues had two stages of denaturation with the following parameters: for bulb nodules: Td(VIII) = 99.4 degrees C, Qd(VIII) = 62.3 +/- 0.8 J/g DNA, Td = (IX) = 106 degrees C, Qd = 28.8 ; 0.4 J/g DNA; and for hippocampus: Td(VIII) = 95 degrees C; Qd(VIII) = 62.0 +/- 9 J/g. Td(IX) - 107 degrees C; Qd(IX) = 29.0 +/- 4.5 J/g DNA. It was established that, along with denaturation of cytoplasmatic structures, nonhistone, nuclear proteins and damage to the nuclear matrix, toluene caused the redistribution of heat between endotherms IX, VIII, VIII(I) connected with the infolding of chromatin loops and 30-nm fibers. It is supposed that toluene causes the damage to the genetic material due to not only its oxidative influence on chromatin DNA but also the disturbance of nuclear matrix structural organization and partial denaturation of nuclear proteins of non-histone origin. PMID- 16358796 TI - [A comparative study of the effects of laser and light-emitting diode irradiation on the wound healing and functional activity of wound exudate leukocytes]. AB - The effects of coherent He-Ne laser and non-coherent light-emitting diode radiation on rat skin wound healing and functional activity of wound excudate leukocytes were compared. A comparative pathomorphological analysis showed that the He-Ne laser and light-emitting diode irradiation stimulated the transition of the inflammatory phase of the wound healing into the reparative (proliferative) and scarring phases sequentially. It was also detected that the functional activity of leucocytes changed in a dose-dependent manner. The leukocyte activity was found to be similar in the groups with laser and light-emitting diode irradiation. Thus, we can conclude that coherent laser and non-coherent light emitting diode radiation have very close effects on wound healing and activity of wound exudate leukocytes, and coherence is not required for this activity. PMID- 16358798 TI - [Study of beta-turns in globular proteins]. AB - The formation of beta-turns in globular proteins has been studied by the method of molecular mechanics. Statistical method of discriminant analysis was applied to calculate energy components and sequences of oligopeptide segments, and after this prediction of I type beta-turns has been drawn. The accuracy of true positive prediction is 65%. Components of conformational energy considerably affecting beta-turn formation were delineated. There are torsional energy, energy of hydrogen bonds, and van der Waals energy. PMID- 16358797 TI - [Quercetin and hesperidin decrease the formation of nitric oxide radicals in rat liver and heart under the conditions of hepatosis]. AB - The formation of nitric oxide radicals in the organs of Wistar rats was studied by the ESR method using ferrous-diethyldithiocarbamate complexes as an NO spin trap. It was found that, under the conditions of CCl4-induced hepatosis, a 4.32 5.46-fold increase in the concentrations of NO radical spin adducts in the samples of heart and liver, correspondingly, occurs. After preliminary intraperoral introduction of quercetin and hesperidin, this effect was not observed. Thus, the flavonoids quercetin and hesperidin exert a distinct antioxidant action, substantialy suppressing the hepatosis-induced increase in the intensity of NO radical generation. PMID- 16358799 TI - Evidence-based dentistry. PMID- 16358800 TI - The "bag o' meds": clinical implications of common patient medications. PMID- 16358801 TI - The causes, prevention, and clinical management of broken endodontic rotary files. AB - Complications can occur during many dental procedures. The prepared clinician responds by either correcting the problem during treatment, or, ideally, preventing the problem from occurring in the first place. In endodontic treatment separated rotary Ni-Ti files are a common procedural problem. Through understanding that the main causes of file breakage are cyclic fatigue and torsional stress, a dentist can best prevent this occurrence by using hand files before rotary files, creating a straight-line (glide path) access into a canal, and preflaring the coronal portion before using rotary files in the apical third of the canal. In addition, using an up and down motion with the electric slow speed handpiece (not allowing the file to bind within the canal) will significantly reduce the incidence of file breakage. If a file does break, successful removal primarily depends on the location of the file in the canal rather than the specific technique employed for removal. A case does not necessarily fail if the separated file cannot be removed. The prognosis when file separation occurs can still be favorable, especially if care was taken to reduce the critical concentration of canal debris with hand instrumentation and chemical irrigation prior to rotary file insertion. In addition, the introduction of a new CS file design will help the dentist increase the chance of removing the file in the event of breakage. PMID- 16358802 TI - An orthopaedic approach to the diagnosis and treatment of disorders of the temporomandibular joint. PMID- 16358803 TI - Replacing hopeless retained deciduous teeth in adults utilizing dental implants: concepts and case presentation. PMID- 16358804 TI - Developing optimal tissue profiles implant-level provisional restorations. PMID- 16358805 TI - Management of the terminal dentition with a bar-supported overdenture and a fixed overdenture: a case report. PMID- 16358806 TI - Aesthetic veneer restoration of an anterior dentition postorthodontics complicated by ankylosis. PMID- 16358807 TI - Demystifying electrosurge. PMID- 16358808 TI - Aesthetic post and core to complement the all-ceramic crown. PMID- 16358809 TI - The use of diode lasers in periodontal therapy: literature review and suggested technique. PMID- 16358810 TI - Intentional extraction and reimplantation of the same tooth. PMID- 16358811 TI - Selecting and using personal protective equipment. PMID- 16358812 TI - The courage to lead. PMID- 16358813 TI - Patient communication: moving beyond 2 tin cans and a string. PMID- 16358814 TI - Co-expression of p16INK4A and laminin 5 by keratinocytes: a wound-healing response coupling hypermotility with growth arrest that goes awry during epithelial neoplastic progression. AB - The replicative lifespan of human keratinocytes in culture is restricted by a telomere-unrelated induction of p16INK4A (p16) and p14ARF. We have found that, in vivo, p16 is expressed by epidermal and oral keratinocytes at the migrating fronts of healing wounds and at the stromal interface of severely dysplastic and early invasive lesions and that such cells also invariably display increased expression of Laminin 5 (Lam5). In culture, p16 and Lam5 are coexpressed in keratinocytes at senescence, at the edges of wounds made in confluent cultures, and when cells are plated on dishes coated with the gamma2 precursor form of Lam5 (Lam5gamma2pre). Lam5/p16 coexpression in all three in vitro settings is associated with directional hypermotility and growth arrest. Hypermotility and growth arrest are uncoupled in p16- and p14ARF/p53-deficient keratinocytes and squamous cell carcinoma (SCC) cells; such cells become hypermotile is response to Lam5gamma2pre but do not growth arrest. Thus, the Lam5/p16 response is activated in normal wound healing, causing growth arrest of migratory keratinocytes that lead wound reepithelialization. This response also becomes activated at a critical stage of neoplastic progression, acting as a tumor suppressor mechanism. Rare premalignant cells that lose p16 remain motile and proliferative, thereby resulting in invasive growth as SCC. PMID- 16358815 TI - Animal models of melanoma. PMID- 16358816 TI - Telomere-dependent chromosomal instability. AB - Telomeres are specialized DNA-protein structures at the ends of the linear chromosomes. In mammalian cells, they are composed of multifold hexameric TTAGGG repeats and a number of associated proteins. The double-stranded telomeric DNA ends in a 3' single stranded overhang of 150 to 300 base pair (bp) which is believed to be required for a higher order structure (reviewed in (Blackburn, 2001)). One important model is that the telomeres form loop structures, the T loops, and by invasion of the 3' overhang into the duplex region of the double stranded part protect the DNA against degradation and hinder the cellular machinery to recognize the ends as broken DNA, thus providing chromosomal integrity (Griffith et al, 1999). If telomeres become critically short they loose their capping function, become sticky, and are prone to illegitimate chromosome end-to-end fusions. The resulting dicentric chromosomes are highly unusable and because of bridge-fusion-breakage cycles they give rise to chromosomal translocations, deletions, and amplifications. Thus, critically short telomeres are thought to be responsible for the onset of genomic instability. In addition, we provide evidence that in a length-independent manner telomeres can confer to genomic instability by forming telomericaggregates which through chromosomal dys locations contribute to chromosomal aberrations. PMID- 16358817 TI - CLIC4, an intracellular chloride channel protein, is a novel molecular target for cancer therapy. AB - Chloride intracellular channel (CLIC)4 is a p53- and tumor necrosis factor alpha (TNFalpha)-regulated chloride channel protein that is localized to the mitochondria and cytoplasm of mouse and human keratinocytes. CLIC4 protein increases in differentiating keratinocytes and in keratinocytes exposed to DNA damaging agents and metabolic inhibitors. Increasing CLIC4 levels by transduction of recombinant CLIC4 causes apoptosis. CLIC4 translocates to the nucleus under a variety of conditions of cell stress, and nuclear CLIC4 is associated with cell cycle arrest and accelerated apoptosis. Reduction of CLIC4 and several other CLIC family members by expressing a doxycycline-regulated CLIC4 antisense also causes apoptosis in squamous cancer cell lines. Expressing antisense CLIC4 in tumors derived from transplanting these cells into nude mice inhibits tumor growth, increases tumor apoptosis, and reduces tumor cell proliferation. Co administration of TNFalpha intraperitoneally enhances the tumor-inhibitory influence of CLIC4 antisense expression. Together, these results suggest that CLIC4 is important for keratinocyte viability and may be a novel target for anti cancer therapy. PMID- 16358818 TI - The importance of being big. AB - The ultimate stem cell, the oocyte, is frequently very large. For example, Drosophila and Xenopus oocytes are approximately 10(5) times larger than normal somatic cells. Importantly, once the large oocytes are fertilized, the resulting embryonic cells proliferate rapidly. Moreover, these divisions occur in the absence of cell growth and are not governed by normal cell cycle controls. Observations suggest that mitogens and cell growth signals modulate proliferation by upregulating G1-phase cyclins, which in turn promote cell division. Like embryonic cells, the proliferation of cancer cells is largely independent of mitogens and growth factors. This occurs, in part, because many proteins that are known to modulate G1-phase cyclin activity are frequently mutated in cancer cells. Interestingly, we have found that both the expression and the activity of G1-phase cyclins is modulated by growth rate and cell size in yeast. These and other data suggest that proliferative capacity correlates with cell size. Thus, a major goal of our laboratory is to use yeast to investigate the relationship between proliferation rate, G1-phase cyclins, growth rate, and cell size. The elucidation of this relationship will help clarify the role of cell size in promoting proliferation in both normal and cancer cells. PMID- 16358819 TI - Normal human melanocyte homeostasis as a paradigm for understanding melanoma. AB - Melanocytes, after cell division, separate and migrate along the basement membrane; they extend their dendrites and establish multiple contacts with keratinocytes. Once adhesion is established, keratinocytes control melanocyte growth and expression of cell surface receptors. Most melanomas arise within the epidermis (melanoma in situ) and then invade across the basement membrane. These melanoma cells escape from control by keratinocytes through five major mechanisms: (1) downregulation of receptors important for communication with keratinocytes such as E-cadherin, P-cadherin, and desmoglein, which is achieved through growth factors such as hepatocyte growth factor, platelet-derived growth factor, and endothelin-1 produced by fibroblasts or keratinocytes; (2) upregulation of receptors and signaling molecules important for melanoma cell melanoma cell and melanoma cell-fibroblast interactions such as N-cadherin, Mel CAM, and zonula occludens protein-1; (3) deregulation of morphogens such as Notch receptors and their ligands; (4) loss of anchorage to the basement membrane because of an altered expression of cell-matrix adhesion molecules; (5) increased elaboration of metal-loproteinases. Thus, investigating normal melanocyte homeostasis helps us to better define how melanoma cells escape the microenvironment created by epidermal keratinocytes and how they develop new cellular partners in fibroblasts and endothelial cells, which support their growth and invasion. PMID- 16358820 TI - [Overview of pandemic influenza]. AB - Influenza virus has a unique characteristics of annual epidemics of acute respiratory disease with attack rate of 10%-30% of the population. It is also the classical emerging infectious disease causing global pandemics when new antigenic shift occur. This antigenic shift is the key to its ability to evoke periodic pandemics, and it has caused at least 3 pandemics in 20th century. I reviewed these 3 pandemics in their natural courses and the epidemiology of the recent emerging influenza A viruses, especially the H5 and H7 subtypes. I described the epidemics of these viruses in human population and why we should be prepared to these viruses. PMID- 16358821 TI - [Modelling the impact of pandemic influenza]. AB - The impact of the next influenza pandemic is difficult to predict. It is dependent on how virulent the virus is, how rapidly it spreads from population to population, and the effectiveness of prevention and response efforts. Despite the uncertainty about the magnitude of the next pandemic, estimates of the health and economic impact remain important to aid public health policy decisions and guide pandemic planning for health and emergency sectors. Planning ahead in preparation for an influenza pandemic, with its potentially very high morbidity and mortality rates, is essential for hospital administrators and public health officials. The estimation of pandemic impact is based on the previous pandemics--we had experienced at least 3 pandemics in 20th century. But the epidemiological characteristics--ie, start season, the impact of 1st wave, pathogenicity and virulence of the viruses and the primary victims of population were quite different from one another. I reviewed methodology for estimation and modelling of pandemic impact and described some nations's results using them in their national preparedness plans. And then I showed the estimates of pandemic influenza impact in Korea with FluSurge and FluAid. And, I described the results of pandemic modelling with parameters of 1918 pandemic for the sake of education and training of the first-line responder health officials to the epidemics. In preparing influenza pandemics, the simulation and modelling are the keys to reduce the uncertainty of the future and to make proper policies to manage and control the pandemics. PMID- 16358822 TI - [The preparedness plan for influenza pandemic]. AB - Influenza A viruses periodically cause worldwide epidemics, or pandemics, with high rates of illness and death. A pandemic can occur at any time, with the potential to cause serious illness, death and social and economic disruption throughout the world. Historic evidence suggests that pandemics occurred three to four times per century. In the last century there were three influenza pandemics. The circumstances still exist for a new influenza virus with pandemic potential to emerge and spread. The unpredictability of the timing of the next pandemic is underlined by the occurrence of several large outbreaks of highly pathogenic avian influenza since the early 1980s. In 1999, the World Health Organization published the Influenza pandemic plan. The role of WHO and guidelines for national and regional planning. And in 2005, WHO revised the global influenza preparedness plan for new national measures before and during pandemics. This document outlines briefly the Korean Centers for Disease Control's plan for responding to an influenza pandemic. According to the new pandemic phases of WHO, we set up the 4 national levels of preparedness and made guidelines for preventing and control the epidemics in each phase. And also we described the future plans to antiviral stockpiles and pandemic vaccine development. PMID- 16358823 TI - [Socioeconomic differentials in health and health related behaviors: findings from the Korea Youth Panel Survey]. AB - OBJECTIVE: This study examined the socioeconomic differentials for the health and health related behaviors among South Korean middle school students. METHODS: A nationwide cross-sectional interview survey of 3,449 middle school second-grade students and their parents was conducted using a stratified multi-stage cluster sampling method. The response rate was 93.3%. The socioeconomic position indicators were based on self-reported information from the students and their parents: parental education, father's occupational class, monthly family income, out-of-pocket expenditure for education, housing ownership, educational expectations, educational performance and the perceived economic hardships. The outcome variables that were measured were also based on the self-reported information from the students. The health measures included self-rated health conditions, psychological or mental problems, the feelings of loneliness at school, the overall satisfaction of life and the perceived level of stress. The health related behaviors included were smoking, alcohol drinking, sexual intercourse, violence, bullying and verbal and physical abuse by parents. RESULTS: Socioeconomic differences for the health and health related behaviors were found among the eighth grade boys and girls of South Korea. However, the pattern varied with gender, the socioeconomic position indicators and the outcome measures. The prevalence rates of the overall dissatisfaction with life for both genders differed according to most of the eight socioeconomic position indicators. All the health measures were significantly different according to the perceived economic hardship. However, the socioeconomic differences in the self rated health conditions and the psychosocial or mental problems were not clear. The students having higher socioeconomic position tended to be a perpetrator of bullying while those students with lower socioeconomic position were more likely to be a victim. CONCLUSIONS: The perceived economic hardships predicted the health status among the eighth graders of South Korea. The overall satisfaction of life was associated with the socioeconomic position indicators. Further research efforts are needed to explore the mechanisms on how and why the socioeconomic position affects the health and health related behaviors in this age group. PMID- 16358824 TI - [Heavy metal as risk factor of cardiovascular disease--an analysis of blood lead and urinary mercury]. AB - OBJECTIVES: We wanted to investigate the relationship between heavy metal, especially lead and mercury, to the blood pressure and cholesterol level in children. METHODS: This study was undertaken in three primary schools and the study subjects were a total of 274 children. The lead in the blood and the urine mercury were analyzed by performing atomic absorption spectroscopy. RESULTS: All of participants' blood lead levels and urine mercury concentrations were below the suggested level of concern according to the criteria of the CDC and ATSDR. We found no significant correlation between lead, mercury and the blood pressure. The blood lead level did not show any relationship with the blood pressure and cholesterol. However, the urine mercury levels were associated with the serum cholesterol. CONCLUSION: Our study suggests that mercury can induce an increase of cholesterol as a risk factor of myocardial infraction and coronary/cardiovascular disease. PMID- 16358825 TI - [Out-of-pocket health expenditures by non-elderly and elderly persons in Korea]. AB - OBJECTIVES: The purpose of this study was to determine the impact of the sociodemographic and health characteristics on the out-of-pocket health spending of the individuals aged 20 and older in Korea. METHODS: We used the data from the 2001 National Public Health and Nutrition Survey. The final sample size was 26,154 persons. Multiple linear regression models were used according to the age groups, that is, one model was used for those people under the age of sixty-five and the other was used for those people aged sixty-five and older. In these analyses, the expenditures were transformed to a logarithmic scale to reduce the skewness of the results. RESULTS: Out-of-pocket health expenditures for those people under the age of 65 averaged 14,800 won per month, whereas expenditures for those people aged 65 and older averaged 27,200 won per month. In the regression analysis, the insurance type, resident area, self-reported health status, acute or chronic condition and bed-disability days were the statistically significant determinants for both age groups. Gender and age were statistically significant determinants only for the non-elderly. CONCLUSIONS: The findings from this study show that the mean out-of-pocket health expenditures varied according to the age groups and also several diverse characteristics. Thus, policymakers should consider the out-of-pocket health expenditure differential between the elderly and non-elderly persons. Improvement of the insurance coverage for the economically vulnerable subgroups that were identified in this study should be carefully considered. In addition, it is necessary to assess the impact of out-of pocket spending on the peoples' health care utilization. PMID- 16358826 TI - [The association of obesity and left colonic adenomatous polyps in Korean adult men]. AB - OBJECTIVES: We wanted to evaluate the relationship between obesity and left colonic adenomatous polyps in Korean adult men. METHODS: This study was conducted among 575 adults men (aged between 40 and 69), who had colonoscopy done from January to December 2002 during a routine health examination at Health Promotion Center, Ulsan University Hospital. The patients' colons were examined up to splenic flexure by using fiberoptic colonoscopy. A questionnaire survey on behavioral factors and physical measurements were also done. The body mass index (BMI) and waist-hip ratio (WHR) were used as the indices of obesity. The BMI was categorized into three levels: normal (BMI < or = 22.9), overweight (23 < or = BMI < or = 24.9), and obese (BMI > or = 25.0). The WHR was categorized into four levels with cut-off points at the 30th, 60th, and 90th percentile of the control group. Age, education, smoking, alcohol use and exercise were controlled for by performing multiple logistic regression analysis. RESULTS: There were 99 cases of colonic adenomatous polyps. Four hundred seventy six subjects with normal colonoscopy findings served as the control. The BMI and WHR were associated with the adenomatous polyps (odds ratio, 1.81 [95% CI=1.02-3.19] for a BMI > or = 25.0 as compared with a BMI < or = 22.9, odds ratio, 3.94 [95% CI = 1.77-8.77] for a WHR > or = 0.95 as compared with a WHR < or = 0.86). The BMI was not associated with the risk of adenomatous polyps after additional adjustment was made for the WHR, but the association between the WHR and adenomatous polyps was still positive and independent of the BMI (odds ratio, 4.15 [95% CI=1.63-10.59]). CONCLUSIONS: The results support that obesity, and particularly abdominal obesity, can be associated with an increased risk of incurring colonic adenomatous polyps. PMID- 16358827 TI - [Alibi verification and the PCR method to estimate the source of epidemic for a few notified cases of S. sonnei]. AB - OBJECTIVES: A few culture-confirmed cases of S. sonnei have been notified from Korean hospitals. The source of epidemic can't be firmly determined in such cases because of the rarity of this illness in the local communities and the timing of the outbreaks. The objective of this study is to estimate the source of epidemic by investigating the patients' lifestyles. METHODS: Alibi verification was used to access the presumed source of the epidemic. PCR (Polymerase Chain Reaction) was used to rapidly detect the genes of Shigella in water specimens. RESULTS: The common lifestyle trait among the Shigella-infected patients was connected with Mt. Martyr in J city, Korea. The first patient's son had gone on a pilgrimage to Mt. Martyr with 41 friends and he had only eaten rice cakes on April 5th; the second patients had visited Mt. Martyr with their mother for a picnic on April 12th; the third patient had visited Mt. Martyr with 22 friends for a pilgrimage and the patient had only drunk holy water on April 13th. Therefore, the holy water of Mt. Martyr was reckoned to be the source of the epidemic. PCR detected the genes of Shigella two days before the S. sonnei was confirmed. CONCLUSION: The patients' lifestyles for 7 days before the onset of symptoms should be determined in terms of time, place and contacted people to find the source of infection when cases with food poisoning are seen in the hospital setting. PMID- 16358828 TI - [Epidemiologic study of Clonorchis sinensis infestation in a rural area of Kyongsangnam-do, South Korea]. AB - OBJECTIVES: To investigate the prevalence of Clonorchis sinensis infestation and to determine the associated risk factors among a population in a part of the rural area of Gyeongsangnam-do, Korea, which is an area known to have a high mortality from liver cancer and a high infection of C. sinensis. METHODS: The study population consisted of those people who have lived in rural areas and who were over 40 years old. This study was performed in 5 areas during the period from 1999 to 2003. Informed consents were obtained from the 2,381 study participants, and these people were interviewed about their life style habits with using the structured questionnaire that was administered by trained staff members. The subjects underwent blood sampling and their stool specimens were examined by using the Formalin-ether sedimentation technique. RESULTS: Among the study subjects (N=2,381), the positivity for C. sinensis in the stool was 34.4% (95% CI= 36.3-42.5); it was 39.4% (95% CI=36.3-42.5) in the males and 30.9% (95% CI=28.5-33.3) in the females. The positivity for C. sinensis was associated with current alcohol drinking (odds ratio=1.8, 95% CI: 1.5-2.1) and raw fish consumption (odds ratio 1.2, 95% CI: 0.9-1.6). CONCLUSION: The present study confirmed the high prevalence of C. sinensis in the study subjects. It is necessary to establish consistent medical management and education programs for the treatment and prevention of C. sinensis infestation in the rural inhabitants of Korea. PMID- 16358829 TI - [Seroprevalence of measles antibody and its attributable factors in elementary students of routine 2-dose schedule era with vaccination record]. AB - OBJECTIVES: We investigated the seroprevalence of the measles antibody and its attributable factors for the students who underwent routine 2-dose Schedule Era. METHODS: The subjects were 996 students of the national measles seroepidemiologic study in December 2000 who had vaccination records. We conducted a questionnaire survey and we performed serologic testing for the measles-specific IgG by using an enzyme linked immunosorbent assay. RESULTS: The coverage for the first dose of the MMR vaccination at 12-15 months of age was 95.1% and the coverage for the second dose of MMR at 4-6 years of age was 35.0%. The proportion of subjects undergoing 2-doses of MMR decreased as the age of the subjects increased. The seropositive rate of the measles antibody was significantly high in the second dose vaccinees (93.5% in the second dose group, 84.7% in the non-second dose group, p<0.001) and it was 72.0% in the 0-dose group, 85.4% in the 1-dose group and 93.7% in the 2-dose group (p<0.001). Two point eight percent of the subjects had a past history of measles infection. On the multiple logistic regression analysis, the first and second dose (odds ratio, 8.54; 95% CI.=3.05-23.91), the first dose (odds ratio, 3.06; 95% CI.=1.20-7.81) and the outbreak in the year 2000 (odds ratio, 1.89; 95% CI.=1.24-2.88) were the significant factors for the seropositivity. CONCLUSIONS: Maintaining high coverage with a 2-dose vaccination program would be the decisive factor to prevent an outbreak of measles and to eliminate measles in Korea. PMID- 16358830 TI - [Folate and homocysteine levels during pregnancy affect DNA methylation in human placenta]. AB - OBJECTIVES: DNA methylation is one of the best characterized epigenetic mechanisms that play a regulatory role in genome programming and imprinting during embryogenesis. In this present study, we investigated the association between DNA methylation in the human placenta and the maternal folate and homocysteine concentrations on the Methylenetetrahydrofolatereductase (MTHFR) genetic polymorphism during pregnancy. METHODS: We investigated 107 pregnant women who visited Ewha Woman's University Hospital for prenatal care during their 24-28 weeks-period of gestation. During the second trimester, we measured the serum homocysteine and folate concentrations. The MTHFR 677 genetic polymorphism was determine by performing PCR-RFLP assay. The expression of DNA methylation in the human placentas was estimated by using immunohistochemistry method. RESULTS: Serum folate was negatively correlated with the serum homocysteine concentration for all the MTHFR genotypes. We found positive correlation between the folate concentrations and the DNA methylation in the human placenta (p<0.05). An increasing concentration of homocysteine was associated with reduced DNA methylation in the human placenta. The coefficient value was -2.03 (-3.77, -0.29) on the regression model (p<0.05). CONCLUSION: These findings suggest that the maternal folate and homocysteine levels along with the MTHFR 677 genetic polymorphism during pregnancy affect the DNA methylation in the human placenta. PMID- 16358831 TI - [Reliability of education and occupational class: a comparison of health survey and death certificate data]. AB - OBJECTIVES: This study was done to evaluate the reliability of education and occupational class between using the health survey and the death certificate data. METHODS: The 1998 National Health and Nutrition Examination Survey (NHANES) was conducted on a cross-sectional probability sample of South Korean households, and it contained unique 13-digit personal identification numbers that were linked to the data on mortality from the Korean National Statistical Office. The data from 263 deaths were used to estimate the agreement rates and the Kappa indices of the education and occupational class between using the NHANES data and the death certificate data. RESULTS: The simple and weighted Kappa indices for education were 0.60 (95% CI=0.53-0.68) and 0.73 (95% CI=0.67-0.79) respectively, if the educational level was grouped into five categories: no-formal-education, elementary-school, middle-school, high-school and college or over. The overall agreement rate was 71.9% for these educational groups. The magnitude of reliability, as measured by the overall agreement rates and Kappa indices, tended to increase with a decrease in the educational class. The number of non-educated people with using the death certificate data was smaller than that with using the NHANES data. For the occupational class (manual workers, non-manual workers and others), the Kappa index was 0.40 (95% CI=0.30-0.51), which was relatively lower than that for the educational class. Compared with the NHANES, the number of non manual workers for the deceased who were aged 30-64 tended to be increased (8 to 12) when using the death certificate data, whereas the number of manual workers tended to be decreased (59 to 41). CONCLUSIONS: The socioeconomic inequalities in the mortality rates that were based on the previous unlinked studies in South Korea were not due to a numerator/denominator bias. The mortality rates for the manual workers and the no-education groups might have been underestimated. PMID- 16358832 TI - [Biosafety of microbiological laboratories in Korea]. AB - OBJECTIVES: The biosafety level (BSL) practiced in microbiology laboratories in Korea according to the laboratory biosafety manual published by the World Health Organization (WHO) was evaluated using the data obtained by a survey. METHODS: Under the advise of Clinical Laboratory Physicians, 144 types of microorganisms were screened based on the guidelines of biosafety in microbiological and biomedical laboratories published by the US Center for Disease Control and Prevention and classified into 1-4 risk groups. A questionnaire containing 21 questions in 5 areas was developed using the biosafety manual by published WHO. Of the 1,876 different organizations sent the survey, 563 responded to the survey (response rate: 30.0%). The species of microoganisms handled by as well as the biosafety level in microbiology laboratories were analyzed. RESULTS: There were 123 species of microorganisms handled in microbiology labs in Korea. The BSL required in 512 microbiology labs was answered by the survey responders as the first grade in 33 labs (6.4%), 2nd in 437 (85.4%), 3rd in 42 (8.2%), and 4th in none. The average number of items satisfied was 12.2, showing only a 57.9% satisfactory rate and normal distribution. CONCLUSIONS: The state of overall observance of BSL in most microbiology labs of Korea was evaluated as lagging compared with the standard set up by WHO. Therefore, the Korean government need to produce and distribute a biosafety manual in microbiology laboratories and make efforts to prevent this threat through measures such as training in biosafety in microbiology labs. PMID- 16358833 TI - [Epidemiologic investigation on an outbreak of salmonellosis in Yeongcheon-si, 2004]. AB - OBJECTIVES: An outbreak of salmonellosis occurred among the students and staff of D primary school in Yeongcheon-si, 2004. This investigation was carried out to prevent any recurrence of this outbreak and to study the infection source and transmission of the salmonellosis. METHODS: The authors conducted a questionnaire survey among 1,205 students and staff members from D primary school about the ingestion of the school lunch and drinking water, and the manifestation of their symptoms. The author examined rectal swabs, the tap water and microorganism cultures, and we also investigated the dining facility and water supply facility. RESULTS: The diarrheal cases were defined as the confirmed cases or the persons who had diarrhea more than one time with symptoms such as fever, vomiting and tenesmus. The diarrheal attack rate was 28.0%. Ingestion of fried bean curd with egg had a significantly high association with the diarrheal attack rate (p<0.05), and the relative risk of the fried bean curd with egg was 10.68 (95% CI=3.88 29.41), as was determined by logistic regression analysis. The bacterial counts in the tap water of the food preparation room and toilet (first floor) exceeded the maximum permissible counts. S. Enteritidis bacteria were only cultured from the fried bean curd with egg of all the supplied foods on September 3, 2004. CONCLUSIONS: The major cause of salmonellosis was presumed to be the contaminated bean curd via contaminated tap water due to a water leak of a school water pipe. This contaminated bean curd was under prepared, which allowed the S. Enteritidis to survive and multiply prior to its ingestion. PMID- 16358834 TI - [Air pollution and respiratory symptoms of school children in a panel study in Seoul]. AB - OBJECTIVES: The aim of this study was to assess the effect of air pollution on the daily respiratory symptoms of elementary school children in Seoul. METHODS: Using the panel study design, we collected diary data for the children's respiratory symptoms during the 1st day approximately 15th day of April, July, October and December in 2003 among the 2nd and 3rd grade elementary school students. We merged the respiratory symptom data with the ambient air pollution data that was monitored by Ministry of Environment. Using a generalized estimate equation, we evaluated the relationship between the daily symptoms of the subjects and the exposure to air pollution after controlling for various potential confounders. RESULTS: The nitrogen dioxide (NO2) exposure of the current day significantly increased the upper respiratory symptoms (adjusted odds ratio=1.12, 95% CI=1.01-1.24) and the lower respiratory symptoms (adjusted odds ratio=1.18, 95% CI=1.06-1.31) in the elementary school children. The sulfur dioxide (SO2) and carbon monoxide (CO) exposure in the current day was associated with the lower respiratory symptoms (adjusted odds ratio=1.12, 95% CI=1.01-1.25 for SO2; adjusted odds ratio=1.16, 95% CI=1.02-1.32 for CO). CONCLUSIONS: We found that exposure to air pollution affects the daily respiratory symptoms in children. This study suggests that the effect on children's health? due to the short term changes in air pollution levels needs to be considered as an important public health problem. PMID- 16358835 TI - [Factors associated with cancer screening participation, Korea]. AB - OBJECTIVES: We wanted to identify those factors associated with stomach, colon, breast and cervix cancer screening. METHODS: A population-based telephone survey was conducted for 2 weeks (the 9th-23th of July, 2004) by trained interviewers with using a questionnaire. 2,598 respondents (females aged 30 years or over, and the males aged 40 years or over) were selected by random-digit dialing that was based on the 2000 Population and Housing Census. The data on socio-demographic, health behavior and enabling factors were collected. 2,571 respondents were included in analysis. The cancer screening rate was classified into 2 categories: the life time screening rate and the screening rate with recommendations. RESULTS: For the 2,571 respondents, the life time screening rate was as follows: 52.0% (Stomach), 25.3% (Colon), 55.9% (Breast) and 76.8% (Cervix). The screening rate with recommendation was as follows: 39.2% (Stomach), 20.6% (Colon), 42.5% (Breast) and 58.3% (Cervix). On a multiple logistic regression analysis of the life time screening, statistically significant relationships were observed for the screening intention, the health exam, the disease history, the age of the patients and the cancer screening rates. On a multiple logistic regression analysis of the screening with recommendation, statistically significant relationships were observed for the screening intention, the health exam, the age of the patients, the concern about the risk of cancer, the voluntary health insurance for cancer and the cancer screening rates. CONCLUSIONS: The results of this study suggest that the cancer screening intention, the health exam and the age of the patients are the most important factors to participate in life time cancer screening and also screening with recommendations. A positive association was also observed for the concern about the risk of cancer, the voluntary health insurance for cancer. It is hoped that this study will be a base line data for suggesting the representative cancer screening rate in Korea. PMID- 16358836 TI - [Investigation of a series of brucellosis cases in Gyeongsangbuk-do during 2003 2004]. AB - OBJECTIVES: We conducted an investigation on 14 cases of brucellosis in Gyeongsangbuk-do during 2003-2004 to understand the source of infection and the transmission routes of brucellosis. METHODS: The authors visited the each of the health centers and we examined the patients, their written epidemiologic questionnaire and the occurrence of bovine brucellosis. We visited the patients' living and work areas, and we examined their occupations, the date they developed symptoms, the progress of their symptoms, whether or not they were treated, their current status, whether or not they consumed raw milk and raw meat, and if their work was related to cattle breeding and the related details. We reviewed the results of the blood tests and medical records and we examined the cattle's barn. RESULTS: There were 3 patients in 2003 and 11 patients in 2004. All of their brucella antibody titer exceeded 1:160. The patients' symptoms were fever, myalgia, malaise, chills and an influenza-like illness, but the clinical signs were absent on the medical records. Brucella abortus were cultured from 3 of the patients' blood samples. CONCLUSIONS: When the authors discovered the transmission routes, they were divided into 4 different sorts. The first route was related to cattle birth such that patients touched the calves or placentas that were infected with the Brucella species. The second route was related to performing artificial insemination on the cattle and the semen that was used for artificial insemination. The third route was due to the ingestion of raw meat and milk. The last route was due to sexual intercourse between the patients. PMID- 16358837 TI - [Noise in military service--the current problem]. AB - Although new norms and regulations are constantly being introduced, noise problem in the Armed Forces still exists. Excessive noise exposure is an inherent danger of the military training. To achieve combat readiness of the unit, training in a real battlefield conditions is essential. It is difficult to plan military training to avoid excessive noise exposure. It is worth noting that 95.8% recruits during selection process referred to the otolaryngology consultants were rejected due to problems with hearing. Norms and regulations of the Polish Armed Forces limiting exposure to noise with comparison to NATO regulations are discussed. Noise exposure on the target range was evaluated. Infra noise hazards were also pointed out. Disability ruling with regard to service connected hearing impairment was described. PMID- 16358838 TI - [Aspects of biological activity of T and NK lymphocytes in children with adenoid hypertrophy and concomitant otitis media with effusion]. AB - The aim of this study was to inquire whether it has got an influence on the number and activity of peripheral blood T and NK cells. MATERIAL AND METHODS: The examined group consisted of 51 children with adenoid hypertrophy. The first group consisted of 17 children with only one episode of otitis media with effusion (ome). The second group consisted of 14 children with more than 4 episodes of ome and the third group consisted of 20 children with adenoid hypertrophy without ome. Evaluation of percentage of CD4+, CD8+ and NK subsets with co-expression of CD69, HLA-DR molecules in peripheral blood was performed by flow cytometry method. In examined groups with ome the decrease of CD4 number was compensated by increase of the number of NK and CD19 cells. In these groups the increase of CD69 and HLA-DR expression on the examined subsets was observed. The results suggest that in recurrent otitis media with effusion may be one of the factors causing T lymphocytes deficiency. Obtained results might be one of the criteria of using adenoidectomy. PMID- 16358839 TI - [Immunoregulatory disorders in chronic sinusitis in patients with atopy]. AB - Disorders of immunoregulatory activity of T lymphocytes and excessive immunogenic activity of antigen presenting cells are recognised as an important pathogenic elements in allergic diseases. Successful prevention of development of these pathomechanisms determines the effectiveness of administered therapy. The aim of presented investigations was to assess the kind and the degree of immune disorders in the group of 40 patients with chronic allergic sinusitis (CAS). The range of evaluated parameters comprised the indices of immune competence of T cells in microcultures of mononuclear cells isolated from the blood (response to mitogens PHA and Con A, saturation of IL-2 receptors, T-cell suppressive activity -SAT index), indices of immunogenic activity of monocytes (LM index correlated with the ratio of concentrations of IL-1 beta/IL-1ra, concentrations of IL-1beta, IL-1ra, IL-4 and IL-6 in the culture supernatants). The results were compared with the similar estimations performed in the group of 20 healthy blood donors. RESULTS: The obtained results demonstrated significant deficiency of T cell immune competence in the group of patients suffering from CAS (hyperreactivity of non-matured T cells to Con A, decreased saturation of IL-2 receptors, deficient T cell suppressive activity and quantitative deficiency of TCD8 cells). In contrast to that, the immunogenic activity of monocytes was significantly elevated in the group of CAS patients as compared to the control group (significantly higher values of LM index, considerable increased concentrations of IL-1b, IL-4 and IL 6). CONCLUSIONS: The observed immunoregulatory deficiency of T-cell population and increased immunogenic activity of antigen presenting cells (monocytes) in CAS patients seem to suggest that the routine antiinflammatory treatment in these patients should be supplemented with the immunocorrective therapy aimed to restore the deficient thymus-dependent immune competence. PMID- 16358840 TI - [The role of chosen cytokines in etiology of nasal polyps]. AB - Nasal polyposis is a chronic inflammatory disease of the nasal mucus. The pathogenesis of nasal polyps is still not entirely known and has been debated for many years. The aim of the present study was to evaluate the expression of interleukins 2 and 4 in patients with nasal polyps and to compare cytokine profile of patients with allergy versus patients without allergy. MATERIAL AND METHODS: Twelve patients with nasal polyps were selected, six of them allergic to dust and six non-allergic. Patients with allergy were distinguished from those without allergy on the basis of positive allergy skin test to dust and the serum levels of IgE. Blood sample was obtained from patients and examined for the expression of Interleukin 2 and 4 by intracellular staining procedure, after stimulation with PMA (phorbol 12-myristate 13-acetate) and allergen (D. pteronyssimus, Allergopharma). RESULTS: Statistical analysis of 2 groups of patients demonstrated that no significant difference in the interleukins expression in allergic versus non-allergic patients was observed (p > 0.05). In patients who used intranasal local steroid before treatment we observed significant decrease of 11-2 level after allergen stimulation (p = 0.034). CONCLUSION: No differences of expression of Interleukin-4 between allergic versus non-allergic patients suggests that the allergic mechanism does not play an important role in the aetiology and formation of nasal polyps. To confirm this, further research needs to be undertaken. PMID- 16358841 TI - [Nasal polyps is not a homogenous pathology]. AB - Nasal polyposis is still a disease of unknown etiology. Looking at its clinical course it seems to be not homogenous pathology. Inflammation of the mucosa, mainly eosinophilic is probably the most important factor, which leads to the development of nasal polyps. THE AIM OF STUDY: To analyze and compare the history data and eosinophilic inflammation parameters in patients with nasal polyps. MATERIAL AND METHODS: 67 patients with nasal polyposis were included into the study. They were divided into three groups: PP-primary nasal polyps (n = 33 49.2%); A-polyps and bronchial asthma without aspirin intolerance (n = 19-28.4%); ASA-polyps and aspirin-induced asthma (n = 15-22.4%). The history data were taken using patient's questionnaire. To assess the eosinophilic inflammation intensity the blood and nasal eosinophilia and serum eosinophilic cationic protein (ECP) concentration were measured. RESULTS: The average age of all patients was 55 +/- 1.5 (range 32-76), the age distribution was comparable in all groups. The ratio males:females--1.5:1 (PP--2.7:1; A--1.1:1; ASA--0.7:1. The females with nasal polyps much more often suffer from asthma than males (p = 0.036). The age when polyps were first seen was 45.1. The duration of nasal polyps was 10 +/- 10 years (range 0.5-30 years): PP-- 10 +/- 9; A-- 14 +/- 11; ASA-- 7 +/- 7 (A/ASA-- p = 0.045). Period of 6.6 years of rhinitis had preceded the diagnosis of nasal polyposis (PP--7.3; A--7.1; ASA--4.5). Age when asthma was developed (A+ASA groups)--45.5; the duration of asthma was 11 +/- 9 years. 58% of patients had polypectomy/ethmoidectomy (PP--45%; A--60%; ASA--80%). The number of operations per one patient: 1.3 (PP--1.0; A--1.2; ASA--2.0), the most often performed in ASA group (PP/ASA-- p = 0.01 and A/ASA-- p =0.045). The mean interval between polypectomies in all patients--9.1 years (PP--10; A--12, ASA--3.5). Blood eosinophilia: 5.6 +/- 0.4% (PP-- 4.6 +/- 0.5; A-- 6.2 +/- 0.9; ASA-- 7.2 +/- 0.9% and PP/ASA-- p = 0.019; PP/A+ASA-- p = 0.03). Nasal eosinophilia: 18.5 +/- 3% (PP - 15.0 +/- 4; A-- 15.5 +/- 6; ASA-- 30.0 +/- 8% and PP/ASA-- p = 0.01; A/ASA-- p = 0.02). Serum ECP concentration: 24 +/- 2 microg/l. CONCLUSIONS: The patient's history and the intensity of eosinophilic inflammation are not similar in all nasal polyposis patients, that support the classification into PP, A and ASA groups. The most severe clinical picture and the highest values of eosinophilic parameters are observed in ASA group. Aspirin intolerance seems to be unfavourable prognostic factor in nasal polyposis. PMID- 16358842 TI - [Assessment of the correlation between taste and smell functioning]. AB - There are few studies examining relationship between taste and smell. However their clinical unity is more often indicated. Smell perception abnormalities cause modification in taste of food intensity, disabling perception of taste as a consequence of taste's sniffing failure. Thus disturbances of these senses, which often commonly coexist could cause great discomfort in patients, however taste sense is estimated as less important comparing to smell sense. The aim of that study was to compare the taste perception in patients with normal smell function and patients with hyposmia and anosmia. There was analyzed correlation between the patients' subjective estimation of smell and taste senses and results of gustometry and olfactometry examinations. MATERIAL AND METHOD: there were indicated three groups of patients based on smell examination with usage of Elsberga--Levy'ego olfactometr in Pruszewicz modification. The first group included patients with normosmia, the second--patients with hyposmia or anosmia related to every smell, the third--patients with hyposmia on few smells and normal smell feeling of others. The Bornstein's method gustometry modified by us was performed for all groups. Moreover patients presented their subjective estimation of smell and taste senses. All results were statistically analyzed, taking into consideration results with alpha < or = 0.05. RESULTS: All (100%) patients with normosmia correctly identified sour, bitter and salty tastes, while only 53% of patients with hyposmia and anosmia got similar results. However 76.5% of patients from the second group subjectively estimated their sense of taste as a normal perception. CONCLUSIONS: Patients with smell disturbances significantly more often have abnormalities in taste perception than patients with normosmia and their subjective estimation of taste and smell senses is much more different than gustometry and olfactometry results. PMID- 16358843 TI - [Evaluation of the cochlear efferent system in patients with cerebello-pontine angle tumor]. AB - The aim of the study was to estimate a cochlear efferent system in the group of patents with cerebello-pontine angle tumor (CPA). The size of an efferent effect was defined as the difference of the amplitudes in click evoked otoacoustic emissions (CEOAEs) during without contralateral acoustic stimulation (CAS) and with broad-bond noise (BBN) was estimated. The research was also conducted on the influence of tumor dimensions, age, sex, and the hearing level in tonal audiometry on the size of the efferent effect. The research was carried out on a group of 14 patients with cerebello-pontine angle tumor and on an appropriate fourteen-person test group. Having our results analyzed we were able to state that, compared with the test group, in the group of patients with CPA tumor significant reduce suppressions of otoacoustic emission response during CAS occurred both in an ear on the tumor side and in an ear on the healthy side. However, the tumor dimensions, sex, and the hearing level in tonal audiometry seemed not to affect significantly the size of the efferent effect. PMID- 16358844 TI - [Effect of nasal obstruction on subjective assessment of sleep quality by the patients with obstructive sleep apnea]. AB - In 52 patients with Obstructive Sleep Apnea Syndrome (OSAS) and nasal obstruction due to nasal septum deformation, hypertrophy of inferior nasal concha or posttraumatic external nose deformity the evaluation of the day somnolence using the Epworth Scale was carried out. The patients were randomly divided in to two groups. The first group consisted of 40 patients who were subjected to appropriate corrective surgery of the nasal obstruction. The remaining 12 patients constituted the control group. After 3 month each of the patients was asked to fill in the same questionnaire. Among the patients who underwent the surgical procedure day somnolence intensity was decreased approximately 2 times whereas in the control group no significant changes were observed. Results of this study could be crucial for the discussion concerning the complex influence of the nasal obstruction on the pathomechanism and symptomatology of obstructive breathing disorders during sleep. PMID- 16358845 TI - [Rhinitis medicamentosa in cytological assessment of the nasal mucosa]. AB - Long term use of topical vasoconstrictors for the nose may result in rhinitis medicamentosa, the rebound swelling of the nasal mucosa. Usually discovery of any underlying reason for decongestant use is impossible, therefore treatment could be problematic. The aim of this study was to establish the nasal mucosal cytology of patients with rhinitis medicamentosa and evaluation of usefulness of exfoliative cytology in diagnostics of underlying reason of nasal decongestants overuse. All patients had a history of six months regular use of popular intranasal alpha-adrenomimetics. All patients have used them constantly with different frequency in 24 h, but always before sleep. The exfoliative cytology was used as a method of nasal mucosa evaluation. Normal cytograms in rhinitis medicamentosa were in 16.4%. Cytograms of 13.4% patients showed a significant number of eosinophils and 29.9% predominant of squamous metaplasia. The study showed that the majority of cytograms in rhinitis medicamentosa indicates nasal mucosa injury. Method of the exfoliative cytology could facilitate the establishment of the underlying cause of intranasal decongestants overuse. PMID- 16358846 TI - [The prevalence of E6/E7 HPV type 16 in laryngeal cancer and in normal mucosa]. AB - Many authors suggest that HPV infection can play a great role in development of benign and malignant tumours of upper respiratory tract in human. The aim of this study was to determine the prevalence of E6/E7 HPV-16 in laryngeal squamous cell carcinoma, and normal laryngeal mucosa, and to analyse their correlation with sex, lymph node status, primary tumor stage, localization, and histological differentiation. HPV 16 DNA presence was analysed using PCR technique in 72 samples of laryngeal carcinoma and in samples of 50 normal mucosa. Human papillomavirus was detected in 26 (36.1%) of the 72 patients. There was no statistically significant correlation HPV positivity and clinicopathological features of the analysed group. In 5 (10%) of 50 samples of normal mucosal. HPV 16 presence in normal mucosa and in laryngeal squamous cell carcinoma was detected in 3 patients. Our observations suggest that HPV16 can play a role in pathogenesis of laryngeal cancer. PMID- 16358847 TI - [An evaluation of the usefulness of impulse-current stimulation in electrogustometry--preliminary report]. AB - Electrogustometry is a simple and fast method of quantitative taste examination. The direct-current monopolar stimulating equipment was most frequently used until now. Aim of the study was to evaluate usefulness of the new method based on bipolar stimulation with impulse-current Furthermore, the authors wanted to determine taste thresholds using the equipment. MATERIAL AND METHODS: 50 healthy volunteers (25 women and 25 men) in the age of 17-50 were involved. The volunteers had a careful otolaryngological examination. Then the taste thresholds for each quarter of tongue were determined using the new equipment. Additionally, ten of the group of volunteers was examined in accordance to repetition time values. RESULTS: the determined values showed symmetry of taste thresholds, among the range of 4.9-26 microA for anterior 2/3 of the tongue and 41.5-93 microA for the 1/3 posterior. The examination with consideration to repetition time did not show increase of taste threshold after ten minutes stimulation. CONCLUSIONS: in conclusion the new kind of electrogustometry is precision and repeatable taste diagnostic method. Moreover the way of stimulation does not cause the phenomena of fatigue and adaptation. PMID- 16358848 TI - [Impulse noise influence on hearing]. AB - Impulse noise from weapons and explosions produces hearing damage among military service. The discovery of otoacoustic emissions (OAE) has given a new possibility of early diagnosis of noise induced hearing loss. The aim of the study was to assess the effects of exposure to impulse noise on TEOAE, SNR and PTA. MATERIAL AND METHODS: the study comprised 92 (184 ears, aged 18.5-20 years) soldiers, subjected to impulse noise during one year of military service. The soldiers were divided in to three groups according exposure to noise. Group I the highest exposure 2222, 9 kPa2/s, group II--611, 8 kPa2/s and group III--103-109 kPa2/s. The control group consisted of 60 (120 ears) secondary school students. Hearing evaluation was performed before and after one year of military service, using extended pure tone audiometry, tympanometry, TEOAE and SNR. RESULTS: after military service significant deterioration of hearing was observed on average 1-2 dB for 0.125-3 kHz and 4-6 dB for 4-12 kHz for all examined soldiers without differences between groups. TEOAE reductions were registered predominantly at frequencies of 3 and 4 kHz, for both ears and SNR mean values reduction for the frequencies of 2, 3 and 4 kHz. CONCLUSIONS: TEOAE especially in cases where clinical audiometry is less sensitive can monitor small changes in the cochlea. There were no differences between groups and ears. PMID- 16358849 TI - [Videostroboscopy assessment in patients with vocal fold hypertrophy underwent microsurgery]. AB - Chronic hypertrophic laryngitis concerns many people in our population. Their aetiopathogenesis is complicated and usually connected with exogenous harmful factors like occupational or environmental exposure. The basic aim of this research was to asses phonation function of the larynx after microsurgery on the basis of videostroboscopy examination. The second aim was to estimate the surgical treatment effectiveness depending on vocal fold hypertrophy extensiveness. The research was conducted on 50 subjects chosen from the population of patients with vocal fold hypertrophy treated at the Department of Laryngology Silesian Medical Academy in Zabrze from January to December 2003. All the examined patients underwent classical microsurgery. Videostroboscopy examination was done just before microsurgery and 3 months after operation. Examination results in two groups with different extensiveness of vocal fold hypertrophy were compared. The first group consist of patients with minor vocal fold lesions (limited to I of vocal fold length), the second group consist of patients with extensive vocal fold lesions (above I of vocal fold length). The control group presented 30 subjects without any voice disorders who did not use their voice for professional purposes. In both groups (with minor and extensive vocal fold lesions) significant improvement in videostroboscopy parameters was observed. The functional results in the group with minor vocal fold lesions were better after microsurgery, but it turned out that results were not statistically significant. PMID- 16358850 TI - [Evaluation of the equilibrium system in patients with multiple sclerosis based on qualitative assessment with videonystagmography]. AB - Multiple sclerosis (SM) is the most frequent inflammatory-demyelinating disease of central nervous system. The character of SM disease provokes that its most frequent symptoms are vertigo, equilibrium disorders and ataxia. Objective method of vertigo estimation is evaluation of the nystagmus via videonystagmography registration (VNG). This examination allows to simultaneously assess the vertical and horizontal component of the nystagmus. It is considered that mainly the vertical nystagmus is characteristic to equilibrium system impairments of the central nervous system, caused also by SM. The study was carried out on 40 patients (28 women, 12 men) with SM diagnosed as a result of neurological examination. 7 patients (17.5%) suffered from sham - movement vertigo, while 33 patients (82.5%) suffered from instability of posture and walking deviation. The videonystagmography examination resulted in the following: deviation of the eye movement was recorded in 26 patients (65%) during either in saccadic test and in smooth pursuit test, optokinetic nystagmus recorded: dissymetric and variable amplitude result in 24 patients (60%), presence of vertical nystagmus component in 30 patients (75%), positional test: directional-changable nystagmus in 8 patients (20%), pendular nystagmus in 4 patients (10%), presence of vertical nystagmus component in 31 patients (77.5%), caloric test: impairments of one of the labyrinth recorded in 12 patients (30%). SM is still the diagnostic and therapeutic problem. During the mentioned tests we have found the quantitative and qualitative changes in VNG recordings. This may be helpful in SM diagnosis, mainly during its early stages. PMID- 16358851 TI - [Evaluation of cochlear function in children with chronic renal failure]. AB - In present study cochlear function was evaluate in young chronic renal failure (CRF) patients using distortion product otoacoustic emissions (DPOAEs). THE AIM OF STUDY: (a) objective assessment of cochlear function in CRF children, (b) establishing best parameters of ipsilateral stimulation in DPOAEs (DP-gram format) for clinical use. Hearing acuity was assessed in 19 CRF children and 18 healthy children. At first pure tone audiometry and tympanometry were evaluate. Only patients with normal middle ear condition and hearing threshold better than 30 dB HL were put forward further investigations. Objective assessment of cochlear function included DPOAEs using DP-gram format. Our results reviled significantly lower mean DPOAE amplitudes in CRF children when compare to healthy children, at all frequencies measured. Most optimal DP-gram parameters in clinical use seems to be level L1 and L2 equal respectively: 65 and 60 dB SPL; 63 and 55 dB SPL; 59 and 45 dB SPL. PMID- 16358852 TI - [Sudden deafness as a presentation of acoustic neuroma]. AB - Sudden deafness (SD) is thought to be a heterogenic group of disorders as to etiopathogenesis. Acoustic neuroma should always be considered in the differential diagnosis. The authors analysed symptoms occurring in the group of 89 patients, diagnosed with acoustic neuroma. A special attention was paid to the role of sudden deafness as a clinical manifestation of the VIIIth nerve pathology. Progressive hearing loss, tinnitus, headache and sudden deafness were the most common complaints. Sudden deafness was developed by approximately 1 of the investigated patients (24.7%). Progressive hearing loss, tinnitus and sudden deafness were also dominating initial signs. The sudden deafness onset was preceded by the period of progressive hearing loss in 3 patients. In one case episode of SD occurred twice. PMID- 16358853 TI - [Detection of Helicobacter pylori in nasal polyps specimens using urease test GUT plus]. AB - In patients with nasal polyps the colonisation of nasal polyps tissues by Helicobacter pylori was investigated using urease test (GUT plus). In this prospective clinical trials we enrolled 61 subjects who had undergone endoscopic nasal polyps surgery and 30 subjects who had undergone septoplasty (control group) were questioned and biopsy specimens of the removed polyps or the mucosal part of inferior turbinate were tested by the urease test GUT plus. In result Helicobacter pylori was not found in the nasal polyps and mucosal part of inferior turbinate tissue of any of patients including the 43 of 61 patients with nasal polyps and 7 of 30 patients with septum deviation who had symptoms of gastroesophageal reflux. In our study using the urease test GUT plus we were not able to confirm presence of Helicobacter pylori in the nasal polyps tissue. However, further epidemiologic studies using different diagnostic methods would be necessary to confirm presence of Helicobacter pylori as potential underlying pathogenetic mechanism of nasal polyposis. PMID- 16358854 TI - [Assessment of cochlea activity in a group of newborns with central nervous system impairment as an effect of perinatal asphyxia using click-evoked otoacoustic emissions (CEOAEs)]. AB - During adaptation of newborn to extra-uterine environment dramatic changes in functioning in the inner organs and the entire human body is observed. Complicated or premature birth as well as complicated course of a perinatal period may cause reversible or irreversible damage of various tissues, organs or systems, and, consequently, their dysfunction. The most often changes in the central nervous system (CNS) in newborns occurring as an effect of perinatal asphyxia are found to be ischemic-hypoxic encephalopathy (IHE), periventricular leucomalacia (PVL) and intraventricular hemorrhages (IVH). Chronic hypoxia is considered to affect the hearing organ in newborns. Reversible or irreversible changes within the cochlea, brainstem or cortex may result in perceptive hearing losses. The aim of the study was to carry out the objective assessment of the cochlea activity using CEOAEs in neonates with CNS impairment occurring following perinatal asphyxia. To the investigation 16 newborns with IHE, PVL or IVH were included. The control group encompassed the health newborns matched as to the age. Perinatal anamnesis, general pediatric status, results of trans-fontanel ultrasonography and biochemical test results were taken into account in statistical analyses. In all newborns otoscopic examination and CEOAEs after birth and 3 months later were performed. CEOAEs in session 1 were significantly reduced in the investigated group comparing to control babies but 3 months later no differences between groups were observed. The outer hair cells activity in first days of life is reduced in newborns with asphyxia in anamnesis but with time no differences in cochlea development are observed. PMID- 16358855 TI - [Computerized image analysis in recognition and classification of aeroallergens]. AB - In medical practice and research, it would be convenient to receive pollen identification and monitoring results in much shorter time than it comes from human identification. Image based analysis is one of the approaches to an automated identification scheme for pollens grain and pattern recognition on such images is widely used as a powerful tool. PURPOSE OF THE STUDY: The goal of such attempt is to provide accurate, fast recognition and classification and counting of pollen grains by computer system for monitoring. METHODS AND MATERIAL: The isolated pollen grain are objects extracted from microscopic image by CCD camera and PC computer under proper condition for further analysis. RESULTS: The algorithms are based on the knowledge from feature vector analysis of estimated parameters calculated from grain characteristics including morphological features, surface features and other applicable estimated characteristics. Segmentation algorithms specially tailored to pollen object characteristics provide exact descriptions of pollen characteristics (border and internal features) already used by human expert. The specific characteristics and its measures are statistically estimated for each object. Some low level statistics for estimated local and global measures of the features establish feature space. Some special care should be paid on choosing these feature and on constructing the feature space to optimize the number of subspaces for higher recognition rates in low-level classification for type differentiation of pollens grain. CONCLUSIONS: The results of estimated parameters of feature vector in low dimension space for some typical pollen types are presented as well as some effective and fast recognition results of performed experiments for different pollens. The findings show the evidence of using proper chosen estimators of tailored characteristics for good enough classification measures even for low dimensional classifiers for type differentiation of pollens grain. PMID- 16358856 TI - [Examination of upper airway function using the dew point hygrometer with semiconductor detector]. AB - The nasal mucosa with blood capillary network has a remarkable role in respiration process. The most important nose functions are heating and humidifying to optimal level of reaching throat and lungs air and partly absorption of humidity and temperature from expired air. Variations of humidifying and heating processes can invoke some upper airways disorders. The paper presents dew point hygrometer which was specially design for laryngological purposes. The hygrometer can measure dynamic changes of humidity in upper airways. The device is fully automated, easy to operate and can communicate with external personal computer. Database application allows saving patient data with archive examination results and can display them easily. During ongoing clinical tests, still increasing amount of data will allow precisely investigate correlations between humidifying process and some diseases. The main advantage of the device is a short response time on humidity changing. The number of readings (detections) can reach 5 readings per second (slightly depending on humidity level) which is much faster than in available on the market hygrometers with sorption sensors. The paper also presents some results obtained in group of healthy volunteers and one patient with tracheostomy The tests figured out actual humidity in certain parts of upper airways: nose, throat, trachea in breathing cycles under various surrounding conditions. The constructed hygrometer can be used for air humidity measurement in upper airways during some diseases and for evaluation of an influence of some drugs and environmental conditions changing on air upper ways mucosa. PMID- 16358857 TI - [Examination of the temperature stabilization process in the upper airways using a miniaturized electronic thermometer]. AB - Diagnostic and treatment procedures in laryngology demand the knowledge on physiological processes taking place in nasal cavities in the course of breathing. It is of special importance in treatment with pharmacologic agents exerting an influence on nasal blood vessels and in nasal structures operations. The paper presents electronic thermometer which was specially designed for measurements in human higher airways. The system is set of flexible measurement head with miniature thermistor and microprocessor system connected to the personal computer fit out database application for collect and visualization measurement data. The device is safe for medical doctor and patient as well. With patient point of view, the measurements can be done easily and comfortably. The system allows recording dynamic changes of air temperature in upper airways and recalculates average inspired and expired air temperatures. The paper presents also initial results obtained in group of 15 healthy volunteers and 3 with tracheotomy It was made an estimation nasal ability to heating inspired air in particular places of nasal cavity and determined air temperature in nasal cavity outlet. PMID- 16358858 TI - [Subjective and objective assessment of voice quality after surgical treatment of overgrowth changes in vocal folds]. AB - OBJECTIVES: Complex estimation of results of treatment micro surgically of ill larynx with overgrowth changes of vocal folds and comparison of usefulness different methods of research phonation function of larynx, and also estimation of influence disturbances of voice on functional state, emotional and physical ill applying to Clinics. MATERIAL AND METHOD: With researches embraced group 40 ill with hoarseness, of which reason one was organic changes of vocal folds under form of polyps, of vocal nodes, of cyst and of swellings Reincke, attended operational (microsurgery with method Kleinsasser). Voice priced twice before intervention surgical and 3 months after operation. In researches used with scale GRBAS, with estimation laryngostroboscopy of function of vocal folds, analysis acoustic created of voice and Polish version of Voice Handicap Index (VHI). RESULTS: Values p = 0.001 for "G" and p = 0.002 for "B" testify about special usefulness these two values of scale GRBAS in estimation of efficiency used treatments. Value p = 0.0004 obtained in research VHI proves distinct statistical symptomatic between estimation one's own voice and his of influence on life ill before and after operation and about good result of treatment. Statistical analysis examined of acoustic parameters showed on indeed statistical symptomatic difference only in chance of variable RAP (p = 0.03). Greatly usefulness for estimations used treatments showed stroboscope research, where especially character close of glottis (p = 0.003) and changes of mucosal wave (p = 0.008) show distinct dependence statistical difference before and after operation. CONCLUSIONS: In complex estimation of voice ill after operation overgrowth changes of vocal folds greatest usefulness showed research of laryngostroboscopy, Voice Handicap Index (VHI) and qualification of grade-G and breathiness-B in scale GRBAS. Thanks used to method VHI ill oneself priced efficiency effect treatments, and doctor knowing answers of patient on each questions canned properly to steer further treatment. PMID- 16358859 TI - [Treatment of otosclerosis in children]. AB - Between 1994 and 2004 in the ENT Department Silesian Medical University Zabrze 330 stapedotomy were performed in patients suffered from otosclerosis. In this group of patient's 16 stapedotomies in 13 children aged from 9 to 17 years were performed. 4.85% of all the patients with otosclerosis treated in our Department were children. The purpose of this study was to analyze the audiologic results of stapedotomies in children. The postoperative evaluations were performed in the period from 6 months to 8 years after the surgical treatment. In the hearing state evaluation we analyzed the change of the air-bone gap, an average air conduction hearing improvement, the rate of percentage hearing improvement and an average hearing loss by Bell Telephone Laboratories. In all the operated children we observed the decrease of the air conduction hearing level. Stapedotomy was successful (postoperative air bone gap within 10 dB) in 62.5% of the cases. In 5 cases the air bone gap was lower than 20 dB (31.25%). In one case it was greater then 20 dB. The air bone gap closure was at the average of 4.39 dB level. The results were comparable with the other authors. Stapedotomy can be an effective procedure for correcting conducting hearing losses due to the otosclerosis in the pediatric patients. PMID- 16358860 TI - [Laser Nd-Yag assisted myringotomy for secretory otitis media in children]. AB - The aim of the study was assessment of the qualities of ND-YAG laser assisted myringotomy as a treatment for secretory otitis media. Nd-Yag laser myringotomy was performed on 52 children (104 ears) mean age 6.8 years diagnosed with secretory otitis media and recurrent secretory otitis media (14.4%). Myringotomy was performed under general anesthesia using the laser (CWNd-YAG laser 3100 Sharplan). Adenoidectomy alone or with tonsillotomy was performed at the same time. Laser tympanostomies remained patent for 6-35 days. All tympanostomies healed with no noticeable scarring. In 15 ears pressure equalization tubes were inserted after myringotomy with laser. Septoplasty performed in 5 children. Laser myringotomy appears to be a safe, and easy to performed, alternative technique in the treatment of secretory otitis media. PMID- 16358861 TI - [Analysis of pharyngeal phase of swallowing after supracricoid partial laryngectomy with cricohyopexia (CHP) and cricohyoepiglottopexia (CHEP)]. AB - The analysis of efficiency and timing of pharyngeal phase events after supracricoid reconstructive laryngectomy was presented. Material consist of 15 patients after partial laryngectomy with cricohyopexia (CHP) and 19 with cricohyoepiglottopexia (CHEP) in average age of 53 (41-66) years. The video radiographic examination of swallowing was done in the early period (max. 120, average 72 days) and in the late period (min. 10, average 24 months) after surgery. The larynx closure improved and the wide of upper oesophageal sphincter (UES) increased in the late period comparing to first examination. The duration of UES relaxation and the duration of pharyngeal phase were similar to physiological status but the time of larynx closure was longer in both periods. PMID- 16358862 TI - [Distant results of spirometric examinations in patients after supracricoid partial laryngectomies with CHP and CHEP type of reconstruction]. PMID- 16358863 TI - [Hearing evaluation in children during chemotherapy]. AB - Chemotherapy is associated with an increased risk of ototoxic changes. The purpose of our study was to investigate the clinical usefulness of Distortion Product Otoacoustic Emissions (DPOAEs) as early indicator of chemotherapy-induced ototoxicity. Tonal audiometry, emission audiometry and DPOAEs were measured in 7 children with acute lymphoblastic leukemia (ALL). Measurements were performed before and after each protocol of chemotherapy. Cochlear activity was evaluated by recording 2f1-f2 DPOAEs with L1=65 and L2=60 dB SPL. Comparisons of the DP grams amplitudes were performed between baseline measurements and those recorded before and after each chemotherapy course. Our results indicate that DPOAEs measurements are very sensitive on early detection of the changes in cochlear function and are recommended for monitor hearing in patients during chemotherapy. PMID- 16358864 TI - [Chronic sinusitis therapy with antibiotics (axetyl cefuroxym, clarithromycin) and steroid (prednisone)]. AB - We started therapy sinusitis of our patients with antibiotics cefuroxime axetil (Zinnat, GSK), clarithromycin (Klacid Uno, Abbott) and orally given steroid prednisone in one group (A+S) 56 patients. Second group of 60 patients were cured only with antibiotics (A). We compare effects of this therapy. There were 50% totally cured patients in the first group (A+S) and 46.6% cured in the second group. Percentage totally cured patients with (A+S) is 3.4% better that cured only with antibiotics in the same time. It is statistically important. We present benefits for patients who were operated in the next step. Post therapy with the use of antibiotics and steroids there were less bleeding from the mucosal membrane, and there was no edema. It is a good method of therapy if patients have no contraindications. PMID- 16358865 TI - [Correlation between computed tomography and microbiological results in chronic sinusitis]. AB - This article presents the observed correlation between the advancement of inflammatory lesions in paranasal sinuses found on computed tomography and microbiological specimen results in chronic sinusitis. The examined study group consisted of 28 patients, aged 19-59 years, in whom endoscopic sinus surgery was performed. In each patient prior to planned surgery a sinus CT scan was performed the result of which was scored using the Lund-Mackay scale, bacteriological and mycological examinations of nasal mucous secretion were also performed. In the study group a positive correlation between the CT score point sum and the number of pathogen species (regression test: r = 0.46; p = 0.0141) was found. With each additional pathogen species found the CT grading rose by 2.3 points. We also found that in patients with positive bacterial culture the average CT score rose by 4.4 (ANOVA: F(1,23)= 3.69; p = 0.0333). However no rise was found in the case of fungi and no interaction between bacteria and fungi was found. In cases of advanced sinusitis where a high CT lesion score of is found antimicrobial therapy based on exact microbiological diagnostics should be given in the perioperative period. PMID- 16358866 TI - [Effectiveness of pharmacologic therapy combined with hyperbaric oxygen in sensorineural hearing loss following acute acoustic trauma. Preliminary report]. AB - Exposure to loud sounds can cause acute acoustic trauma and permanent sensorineural hearing loss. The aim of the study was to evaluate effectiveness of pharmacological treatment combined with the hyperbaric oxygen (HBO) in the treatment of sensorineural hearing loss following acute acoustic trauma for patients treated in the Department of Otolaryngology of Military Institute of Health Service and Warsaw Center for Hyperbaric Therapy and Wounds Treatment. MATERIAL AND METHODS: Group of 12 patients (22 damaged ears) with acute acoustic trauma (10 men and 2 women), 16-67 years of age, mean age 28 years, was involved in our study. Patients with tinnitus and sensorineural hearing loss of minimum 15 dB at 0.25-8 kHz were included in the treatment group. Hearing gain of minimum 10 dB at 0.25-8 kHz and decrease in the intensity of tinnitus was considered as an improvement. 4 days was the mean time interval between acoustic trauma and starting the pharmacological treatment, 7 days was the mean time interval for the hyperbaric oxygen therapy commencement. RESULTS: Statistically significant difference in Pure Tone Audiometry results obtained before and after the treatment was noted in 4 kHz when considering all damaging factors that caused acoustic trauma and in 6 kHz only for damage resulting form shooting. Statistically significant difference was noted in 6 kHz if the HBO therapy was continued over 10 days (p < 0.00001). Statistically significant difference was noted in 4, 6, 8 kHz when treatment was started within 5 days since the acoustic trauma (p < 0.000001). CONCLUSIONS: Hyperbaric oxygen therapy is the unique method of increasing concentration of oxygen in the inner ear fluids thus facilitates the regeneration process. Hyperbaric oxygen therapy combined with steroids is an effective method of sensorineural hearing loss treatment following acute acoustic trauma. PMID- 16358867 TI - [Progression of larynx and hypopharynx cancer in the last ten years]. AB - Epidemiological analysis of patients with the larynx and hypopharynx cancer, who were treated in the Department of Otolaryngology in Bialystok from 1999 to 2004 was performed. The following aspect were assessed: (1) number of patients, (2) sex, (3) profession and place of living, (4) primary site of the tumour, (5) clinical advancement stage of the disease. The study was of 676 patients. The majority--92.2%--were males but an increase in number of female patients was found. The most common site of the primary tumour was the supraglottic region (56.9%). The analysis showed that the numbers of patients with T3/T4 clinical advancement stage of the tumour was larger (64.9%). PMID- 16358868 TI - [Quality of life for patients with laryngeal cancer]. AB - The number of diagnosed instances of larynx cancer is systematically increasing, especially in men. Treatment, in the majority of cases, is undertaken at an advanced stage of the disease. This is the major factor determining the outcome of therapy and the patients' quality of life. The aim of this paper is to estimate the environmental and occupational risk factors as well as to evaluate the quality of life of patients before they were diagnosed with larynx cancer and after the treatment was completed. The analysis involved 82 patients after total laryngectomy due to squamous cell carcinoma. A vast majority of the patients (96%) abused alcohol and were heavy smokers (95%). 76% were exposed to harmful environmental conditions and 56%--to harmful conditions at workplace. After treatment, which was accepted by over 96% of the patients, many of them quit using alcohol (57%) and quit smoking (73%). Because of difficult economic situation only few of them decided to change their working or living conditions. 47% of patients judged that after the diagnosis and consequently after undergoing complete treatment, the quality of their life deteriorated. Their awareness of the health-threatening factors had however grown. PMID- 16358869 TI - [Evaluation of polypectomy-related changes on olfactory function]. AB - AIM: Aim of this study was to evaluate polypectomy-related changes of olfactory function in early postoperative period in comparison to the preoperative situation. MATERIAL AND METHODS: Olfactory function was assessed in 25 patients of Military Medical Institute using Elsberg-Levy method. Measurements were performed preoperatively and 1 month after polypectomy. RESULTS: Before the polypectomy measurements revealed higher odor thresholds in 18 patients. Postoperatively, decline of odor thresholds was observed in 14 hyposmic patients. CONCLUSION: Present data indicates that olfactory function is related to nasal obstruction resulting from nasal polyposis. The study showed that polypectomy can increase olfactory function in patients with nasal polyps. Olfactometry can help to estimate nasal polyps during and after treatment even in early stage. PMID- 16358870 TI - [New method of biopsy of palatine tonsils and its role in the indication for tonsillectomy (preliminary report)]. AB - In our work we presented a new method of chronic tonsillitis diagnosis and evaluated its usefulness in qualification to tonsillectomy. Oligobiopsies were obtained from right and left palatine tonsils of 36 patients who were qualified to tonsillectomy on the basis of past medical history, otolaryngological examination and laboratory tests findings. Following gauge of biopsy instruments were used: 2 mm, 3 mm, 4 mm. The material obtained at biopsies and tonsillectomies was histopathologically evaluated. Histopathological findings of specimens obtained at oligobiopsies and post tonsillectomies were compared. The presence of interstitial fibrosis, neutrophils infiltrates, hyperplastic changes of lymphoid follicles of palatine tonsils were evaluated in oligobiopsy specimens in comparison to the results of postoperative specimen findings. Full concordance of histopathological findings obtained at oligobiopsy and post tonsillectomy was noted with the use of biopsy instruments of 3 mm and 4 mm gauge. Specimens obtained at oligobiopsies had a great diagnostic value. It was proved that the biopsy performed according to our method has a great diagnostic value and helps surgeon to qualify patient to tonsillectomy. PMID- 16358871 TI - [Self-assessment of the effect of dysphagia on the quality of life in patients after partial laryngectomy for cancer initially located in the supraglottic area]. AB - Dysphagia is frequent in patients after partial laryngectomy for cancer initially located in the supraglottic area. To ensure the best quality of life, establishing how the patient feels after treatment is necessary. Therefore, a self-administered questionnaire was designed to evaluate the dysphagia. 95 patients (75 male and 20 women) who were operated in ENT Department CMUJ between 1998-2004 participated in this study. The MDADI questionnaire in the Polish version was used. The subjective evaluation by the patient of dysphagia is directly related to the size of the tumor and the extent of the partial laryngectomy treatment. Patients, whose tumor was located in supraglottic-glottic area, had subtotal laryngectomy m. Miodonski. Their quality of life was significantly worse due to dysphagia than those with tumors located only in the supraglottic area. Swallowing dysfunctionality was significantly greater after the resection of the hyoid bone. Reconstruction of the base of a tongue by a graft of angiopedunculated submandibular gland diminished the difficulties in swallowing. Moreover, the longer the period after treatment, the higher the quality of life becomes because of the improvement in the swallowing functionality. PMID- 16358872 TI - [Salvage surgery for advanced laryngeal and hypopharyngeal carcinoma]. AB - The aim of this study was to present results of treatment of advanced laryngeal and hypopharyngeal carcinoma in 334 patients, who were treated by primary radiotherapy and salvage surgery in Department of Otolaryngology of the Medical University of Warsaw between 1975 and 1990. Cancer of the larynx was diagnosed in 295 cases and cancer of hypopharynx in 39 cases. Depending on the extend of the tumor different types of surgery were performed. Some of the patients included in the study were treated by jejunal interposition procedures and pectoralis major myocutaneous flap to reconstruct pharyngoesophageal defects. Presence of nodular metastases and the necessity of an extend surgery were the cause of the increase of recovery. The 3-years survival after treatment without recurrence of the disease was:--among patients with laryngeal cancer: 56% (167 patients)--among patients with hypopharyngeal cancer: 38% (15 patients). PMID- 16358873 TI - [The treatment results of regional metastases in advanced laryngeal and hypopharyngeal carcinomas]. AB - Metastases from primary laryngeal and hypopharyngeal cancers to cervical lymph nodes is of great importance in designing treatment schemes. The aim of this study was to explain: (1) Do site of primary tumor, its stage and histological differentiation have significant influence on formation metastases to lymphatic nodes of the neck; (2) Do the lymph nodes metastases influence on results of treatment? MATERIALS AND METHODS: We have analyzed records of 586 patients with advanced cancer of larynx and hypopharynx, who were treated by surgery between 1975 and 1990 in Department of Otolaryngology Medical University of Warsaw The clinical stages of cancer was defined according WHO classification (1987). RESULTS: cancer of the larynx was diagnosed in 477 cases and in 109 cases--of hypopharynx. The lymph nodes metastases were diagnosed in 152 (32%) cases with laryngeal carcinomas and in 97 (89%) cases with hypopharyngeal carcinomas. In every case the presence of unilateral metastases in lymphatic nodes of the neck was the reason of radical neck dissection, in bilateral metastases radical neck dissection was applied at one side of the neck and functional neck dissection on the second side. CONCLUSIONS: Lymphatic metastases in laryngeal cancer are related to: site of primary tumor, extension of primary tumor and histologic differentiation. The metastases to lymph nodes in cancer of hypopharynx are diagnosed in the greater part, independently from progression of tumor. Presence of metastases in lymphatic nodes influence significantly on estimation of the treatment independently from progression of tumor. It was state that the best results give the method of combined treatment: the surgery and postoperative radiotherapy. PMID- 16358874 TI - [Effectiveness of therapeutic (NI, N2) modified radical neck dissection (MRND) in patients with laryngeal cancer]. AB - OBJECTIVE: Efficacy of modified radical neck dissection in treatment of laryngeal cancer with node positive neck. PATIENTS AND METHODS: Retrospective analysis of 311 patients treated with modified radical (86) or radical (225) neck dissection. Recurrence rates, local and regional, after both operations and 5-year disease free survival (Kaplan-Meier method) were analyzed. RESULTS: Regional recurrence rates in N1 group after MRND were 3% and 13% after RND. In N2 group 13% and 16% respectively. 5-year survival after MRND in N1 group was 73% compared with 60% after RND. In N2 group 5-year survival was 56% and 47%. CONCLUSION: We consider MRND as effective treatment method of node positive neck in N1 and N2 stage compared to RND with less morbidity. PMID- 16358875 TI - [Extracapsular spread as an ominous prognostic sign in laryngeal cancer]. AB - OBJECTIVE: Impact of extracapsular spread upon survival and recurrence rates in patients with laryngeal cancer. MATERIAL AND METHODS: Retrospective analysis of 309 patients with pathologically confirmed regional metastases with and without extracapsular spread (144 and 165 patients respectively). To examine impact of clinical variables upon treatment outcomes we used logistic regression (logit) and Kaplan-Meier method to calculate 5-years disease free survival. RESULTS: Extracapsular spread significantly reduced 5-years survival (40% vs 77%; p = 0.005) and increased regional recurrence rates (33% vs 12%; p = 0.0001). CONCLUSION: Authors suggest use of radiotherapy after neck dissection in presence of extracapsular spread. PMID- 16358876 TI - [Squamous cell carcinoma antigen levels in patients with laryngeal cancer]. AB - Squamous cell carcinoma antigen (SCCAg) is one of the most common markers used in diagnosis of head and neck cancer and larynx cancer. We tested correlations between level of SCC Ag and tumor size, presence of lymph node metastasis, clinical advances of tumour and histopathological diagnosis. Pretreatment level of SCC antigen was evaluated in 34 patients with squamous cell carcinoma of the larynx. Microparticle enzyme immunoassay was used to measure the SCCAg level. Elevated SCCAg serum levels were found in 41% of patients. The magnitude of the marker elevations were correlated with lymph node metastases (N0 versus N2, and N1 versus N2). Our date indicate that in patients with larynx cancer SCCAg does not appear to be a sensitive marker in the primary diagnosis. However, seem to be useful marker for monitoring nodal invasion. PMID- 16358877 TI - [Nitric oxide as indicator of paranasal sinusitis]. AB - Squamous cell carcinoma antigen (SCCAg) is one of the most common markers used in diagnosis of head and neck cancer and larynx cancer. We tested correlations between level of SCC Ag and tumor size, presence of lymph node metastasis, clinical advances of tumour and histopathological diagnosis. Pretreatment level of SCC antigen was evaluated in 34 patients with squamous cell carcinoma of the larynx. Microparticle enzyme immunoassay was used to measure the SCCAg level. Elevated SCCAg serum levels were found in 41% of patients. The magnitude of the marker elevations were correlated with lymph node metastases (N0 versus N2, and N1 versus N2). Our date indicate that in patients with larynx cancer SCCAg does not appear to be a sensitive marker in the primary diagnosis. However, seem to be useful marker for monitoring nodal invasion. PMID- 16358878 TI - [Immunological and clinical study on therapeutic efficacy of inosine pranobex]. AB - Many studies in vitro and in vivo have shown immunomodulating and antiviral activities of inosine pranobex. The object of this research was to examine the potential beneficial effects of inosine pranobex (Groprinosin) on immune system in children with cellular immunodeficiency as a prophylaxis of recurrent infections, mainly of viral origin. 50 mg/kg b.w/day of inosine pranobex in divided doses was given to the group of 30 children aged 3-15 years for 10 days in 3 following months. Clinical and immunological investigations were done before and after the treatment. Statistically significant rise of CD3T lymphocytes number (p = 0.02) and in this CD4T lymphocytes number (p = 0.02) as well as statistically significant improvement of their function (p = 0.005) evaluated with blastic transformation method were found. These laboratory findings were parallel to clinical benefits. Control study was performed in the group of children completed by randomization and treated in the same way with garlic (Alliofil). PMID- 16358879 TI - [Exemplary classification of otolaryngologic data using the Kohonen artificial neural network]. AB - An exemplary classification of otolaryngological data with use of the Kohonen artificial neural network is presented along with an evaluation of grouping effectiveness. The study was based on 220 patients with otosclerosis, including 184 women and 36 men, aged 16-68, operated at the Otosurgery Department, Medical University of Lodz in 1984-1992. The neural network distinguished 3 clusters considerably differing in air and bone conduction preoperatively and as a result in the values of postoperative air-bone gap. The effectiveness of classification was also exhibited in congruent with previous knowledge differences between clusters in many other variables not presented to the neural network. PMID- 16358880 TI - [Orbital complications in acute and chronic sinusitis]. AB - Orbital complications in acute and chronic sinusitis occur much more rarely than previously. Orbital and ocular symptoms are also present in chronic sinusitis: mucocele, pyocele and cholesterol granuloma. We describe 33 patients with orbital complications and analyse symptoms, diagnostic procedures and treatment options. The most frequent presenting orbital and ocular symptoms are lid oedema, proptosis, pain in orbita. We got 80% correlation in assessment of bony frame of orbita in CT and MRI with intraoperative view, in 13.3% they were false positive. Pharmacologic treatment alone was used in six patients, and 27 were treated surgically. There was one case of blindness. PMID- 16358881 TI - [Anxiety and depression in patients with laryngeal and hypopharyngeal cancer (preliminary report)]. AB - Occurrence of anxiety and depression was investigated in 76 patients with recognized larynx and/or hypopharynx cancer and with or without alcohol dependence. The patients were treated using total or partial laryngectomy or radiotherapy. Patients were examined three times using questionnaires: SCID, BDI, STAI. The greatest intensity of anxiety was observed before treatment, especially in patients qualified for surgery. After finishing the treatment, independently of the type of operation, intensity of anxiety was lower than in patients treated using radiotherapy In patients after total laryngectomy the greatest intensity of depression was observed 7 days after operation. The mood of these patients has improved significantly before discharge from hospital. In patients treated with partial laryngectomy such great changes of mood were not observed. In patients treated using radiotherapy were stated greater intensity of depression at the end of treatment - despite of initial mood improvement. Patients with alcohol dependence were stated greater intensity of anxiety and depression than in the other patients. Dynamics of the intensity of anxiety and depression in the different diagnostic and therapeutic groups of patients with laryngeal or hypopharyngeal cancer should be done under consideration of their prophylactic and rehabilitative effects. PMID- 16358882 TI - [Analysis of select diagnostic examination results and their connection with cervical vertigo diagnosis]. AB - Cervical spine spondylotic changes are known to cause vertigo, but diagnostic criteria are not established. The aim of present study was to asses diagnostic value of classical radiologic, electronystagmographic (ENG) and transcranial doppler ultrasonographic (TCD) examinations in cervical vertigo diagnostic process. Forty patients with vertigo lasting more than 6 months with radiological evidence of cervical spondylosis participated in the study. All patients had ENG and TCD with neck rotation test. Cervical vertigo was diagnosed in patients with positive neck rotation test in: TCD examination (15% decrease of vertebral artery flow) and/or ENG examination (cervical nystagmus). We diagnosed cervical vertigo in 65% of patients. Multivariate model of logistic regression analysis showed that the best statistical predictors of cervical vertigo were: asymmetry of vertebral artery flow in TCD (OR: 92.2; 95% CI: 6,2-1381) and presence of osteophytes and discopathy in radiological examination (OR: 64.1; 95% CI: 1,3 1107). The transcranial doppler ultrasonographic examinations seems to be useful for diagnosing cervical vertigo. PMID- 16358883 TI - [Kinesitherapy treatment for patients with cervical vertigo (preliminary report)]. AB - The aim of the study was to assess the effectiveness of kinesitherapy in cervical vertigo treatment. Thirty two patients with cervical vertigo, aged 20 to 75, were examined. The patients performed kinesitherapy exercises three times a day for four weeks. The effects of therapy were assessed by everyday task self-control cards and ENG examinations. In the present study, objective improvement was noticed in 18,7% patients. Subjective improvement assessed by the patients everyday task self-control cards reached 62.5%. Kinesitherapy seems to be the good method of treating neck-related vertigo, but it should be confirmed in further study on the large group of patients. PMID- 16358884 TI - [Influence of positional blood flow disorders in the vertebral and basilar arteries on the prevalence of vertigo in patients with cervical spondylosis]. AB - The aim of present study was to characterize the association between prevalence of vertigo and age, grade of radiological changes and positional vertebral (VA) and basilar artery (BA) flow lesion in patients with cervical spondylosis. We examined 80 patients with radiological evidence of cervical spondylosis. Forty patients complained positional vertigo. Patients with neurological symptoms were excluded from the study. ENG was used to exclude patients with another causes of vertigo. TCD evaluations of the distal part of VA and the proximal part of BA were performed with a 2-MHz probe via the suboccipital window. We showed significant and independent association between prevalence of vertigo and age, grade of radiological changes, and positional vertebral and basilar artery flow lesion velocity in multivariate logistic regression model. Spondylotic-induced VA compression may be the reason of decreased blood flow velocity in the basilar artery during head rotation, causing positional vertigo. PMID- 16358885 TI - [Techniques of intranasal drug use]. AB - The aim of the study was to assess the incidence of changes in nasal mucosa in patients treated with long term intranasal corticosteroids and to determinate reasonability of intranasal drops administration in symptomatic treatment of nasal catarrh. The study included the total of 1962 patients. Within this group, 612 patients treated with intranasal corticosteroids underwent full laryngological examination to evaluate the status of nasal mucosa. In the group of 1350 users of web-based Allergologic Courier, who had used intranasal drops because of "runny nose" within last 3 months before the examination, a questionnaire survey was performed. The study results reflect poor knowledge the methods of nasal drops administration among patients, as well as low effectiveness of educational initiatives led by physicians and pharmacists. In 25 persons from 367 examined (6.81%) who used steroid spray locally, and who showed up in laryngological outpatient clinic, nasal mucosal damage was diagnosed for the first time. Mucosal damage was most frequently located in the right nasal septal mucosa--in 21 patients (5.72% of total patients) that was connected with improper nasal inhaler positioning and right hand movements directing it towards septum in the right nasal meatus. Among patients, who have remained under regular laryngological care in outpatient clinic (all of them are instructed of drug administration technique), only 5 persons (2.04%) out from 245 in the study group presented changes in nasal mucosa. No differences between changes occurrence on each side of nasal septum were found. Without proper training, only 8.52% from 1350 examined persons had been taking nasal drops in a proper manner to achieve its effectiveness. PMID- 16358886 TI - [Prognostic significance of depth and mode of invasion of the neoplasm in the presence of micrometastases in laryngeal carcinoma--cytokeratin antigens expression in lymph nodes]. AB - According to the recent reports analyses of characteristic of neoplasm's invasion is one of the most reliable methods of estimation the progress of the changes of tumor and prediction the outcome in patients with laryngeal cancer. Depth and mode of invasion are features which are able to assess the dynamics of the tumor growth quite precisely. In this study it was presented direct relation between morphological features of tumor front as a depth and mode of carcinoma's growth and the probability of micrometastases. The authors have analysed 22 cases of patients who were operated on the laryngeal squamous cell carcinoma in ENT Department Medical University of Lodz in the period of 1998-1999. Features of the TFG and its dependence on lymph node micrometastases, considered in immunohistochemistry with using of panel of cytokeratins (CKs) were analysed. Our study showed that feature such as depth and mode of invasion is very useful in prediction of micrometastases in patients with laryngeal squamous cell carcinoma. PMID- 16358887 TI - [Prognostic significance of clinical and morphological features of the lymph nodes in the presence of micrometastases--cytokeratin antigens expression in lymph nodes in laryngeal carcinoma]. AB - In patients treated surgically due to neoplasm, in lymph nodes can be presented micrometastases, focuses 1-2 mm of carcinoma's cells. Lymph node micrometastases in patients with squamous cell carcinoma of the larynx found node-negative by conventional histology may be discovered by immunohistochemistry, particularly by using antibodies which are reactive with epithelial cells eg. cytokeratin antigens. Then tumors classified as pN0 by routine methods could be reclassified more correctly as pN1. Authors investigated the incidence of micrometastases in the neck dissection specimens originally staged as pN0 from 22 patients with laryngeal cancer treated surgically at ENT Department Medical University of Lodz between 1998-1999, according to the survival, using immunostaining with panel of polyclonal antibodies to cytokeratins. The relationship between micrometastases and clinical (pre- and intraoperative) features of lymph nodes were estimated. PMID- 16358888 TI - [Evaluation of osteomeatal complex in children with chronic sinusitis]. AB - The aim of this publication was estimation of osteomeatal complex and its visualisation by means of CT in children having been treated conservatively and surgically because of sinusitis. There were analyzed 47 children age 4 to 18 years who had been treated in Dept. of Children Otolaryngology, Phoniatry and Audiology of Medical University in Lublin during 2001-2005. It was found, that the causes of sinusitis are disturbances of osteomeatal complex and that the frontal projection in CT investigation is more efficient in preoperative diagnostics than sagittal projection. PMID- 16358889 TI - [Reconstruction of meatal wall with cartilage and perichondrium]. AB - The aim of the study was evaluation of the use of cartilage and perichondrium for reconstruction of meatal wall. In the group of 27 patients transmeatal atticotomy was performed to remove cholesteatoma from epitympanic recess. Auricular or tragal perichondrium with cartilage island were used for reconstruction of meatal wall and tympanic membrane defect. Good closure of the defect was obtained in 24 (89%) patients in 3-6 years follow up. We had one failure and no follow up in 2 patients. Of 7 patients with intact ossicular chain 0-10 dB air-bone gap was obtained in 5 and 11-20 dB gap in 2 patients. Perichondrium and cartilage are very good for reconstruction of meatal wall and drum defect. PMID- 16358890 TI - [Site of origin of paranasal sinus malignancies]. AB - In a group of 124 patients a site of origin of paranasal sinus and nasal cavity malignancies was evaluated on the basis of computed tomography (CT) and on the basis of operative findings. The most common malignant tumor was squamous cell carcinoma (41, 1%) and then cancers of salivary origin (20, 2%). The most common site of origin was the maxillary sinus (53, 2%) and then the nasal cavity (31, 5%). Within the maxillary sinus the most frequent neoplasm was cancer whereas in the nasal cavity melanoma and olfactory neuroblastoma. In three (2, 4%) patients site of tumor origin demonstrated on CT was different form that of operative finding. PMID- 16358891 TI - [Salvage surgery after failure of radiotherapy in laryngeal cancer]. AB - OBJECTIVE: Efficacy of salvage surgery after radiotherapy failure in patients with laryngeal cancer. MATERIAL AND METHODS: Retrospective analysis of 121 patients treated in our clinic between 1980 and 2003 (supraglottis--86 patients, glottis--35). In 70% of patients T-stage was T3 and T4 and regional metastases were present in 56% of cases. We performed statistical analysis of recurrence rates and 5-years survival according to Kaplan-Meier method. RESULTS: Glottic localization (p = 0.02) and extracapsular spread (p = 0.03) were significant factors of final outcome. Local and regional stage was not significant factor of recurrence. Disease free 5-year survival was 53%. CONCLUSION: Salvage surgery is a reasonable method that could save about 50% of patients after radiotherapy failure. PMID- 16358892 TI - [Carbon granuloma--an unwanted effect of laryngeal laser microsurgery]. AB - Over recent twenty years, laser microsurgery has gained great recognition in the treatment of early carcinoma. From 1990 to 2004, more than 800 operations on the larynx were performed using laser microsurgery, including 167 chordectomies for squamous cell carcinoma. In 14 patients who were on the follow-up for 1 to 6 months, we observed a spherical, smooth structure (3-5 mm in diameter) in the middle part of the scar, which imitated local relapse of the neoplastic process. The lesion was removed with CO2 laser. Histopathologic examination revealed, in all cases, the presence of nonspecific, inflammatory process and development of granulation tissue. In only one patient in further follow-up, neoplastic growth recurred. Such lesions were not observed in other than chordectomy laser microsurgical procedures. PMID- 16358893 TI - [Long-term results of stapedectomy in pediatric patients]. AB - Otosclerosis is generally considered as a disease of adults, but the onset of hearing loss can occur in childhood. The purpose of this presentation is to show the effectiveness of stapedectomy in children. Both, short and long-term results are presented. Material is based on 14 pediatric patients, age 7-14 y. 12 of them had bilateral, 2 unilateral hearing loss. Together 26 ears. The diagnostic pattern was as follow: general pediatric and ENT examination, tuning fork testing, pure-tone audiometry (2x), recognition score test, tympanogram and acoustic reflex test, CT scan of the temporal bone. Particular attention was paid to the parents report concerning the episodes of otitis media or frequent upper respiratory tract infections and history of family hearing loss. Average time of hearing loss was noted 3 years before stapedectomy. Pure tone audiometry was performed on frequencies 500, 1000, 2000, 3000 and 4000 Hz. All children had hearing loss with air-bone gap 30 or more dB. 23 ears underwent stapedectomy for otosclerosis with teflon-piston prosthesis insert. Hearing level was based on PTA obtained in 2-3 weeks postoperatively and then 3 and 6 months. PTA hearing improvement was in 22 ears. No hearing loss was noted after operation. A total of 16 ears were available for long-term follow-up (up to 24 months after stapedectomy). The PTA hearing improvement was found to be permanent. PMID- 16358894 TI - [Neuro-otologic findings in multiple sclerosis]. AB - Neuro-otologic examination is very important in evaluation of balance system. Multiple sclerosis (MS) is the most common chronic, debilitating disease characterized by focal demyelinization that develop throughout of the central nerves system at varying time. Patients with MS presented various complaints and signs, so the aim of the study was the neuro-otologic presentation of multiple findings in patients with MS and comparison with reviewing literature. The study was carried out in 32 patients with MS (21 with relapsing-remitting, 9 with secondary progressive, 1 with primary progressive and 1 with progressive/relapsing types) the mean age was 38 +/- 10 years. Clinical neuro otologic examinations were performed in all patients paying special attention to eye movement ability. Vertigo as the first symptom was found in 18.8% of patients but up to 68% of patients would complain this symptom at some point of disease. The hearing impairment and tinnitus complains 6.2% patients, optic neuritis 28% patients and double vision 46.9% of them. A few types of eye movement disorder like abnormalities of fixation, gaze-evoked nystagmus, acquired pendular nystagmus and vertical nystagmus in 46.8% patients were observed. The clinical signs of internuclear ophthalmoplegia were noticed in 2 patients. Disturbance in voluntary gaze either in smooth pursuit test (31.3%) and in saccadic test (43.8%) were seen. The multifocal nature of MS explain why this disease are so commonly subject to defect different part of nerve system. Careful neuro-otologic assessment of every patient with MS is necessary to confirm and monitoring the course of disease. PMID- 16358895 TI - [Stomaplasty for trachoestomal stenosis using rib cartilage--authors method]. AB - Authors present surgical procedure for treatment of tracheostomal stenosis after laryngectomy with use of 7th rib cartilage. Procedure is simple, almost without morbidity, provides good ventilatory and cosmetic results and helps in learning esophageal speech. PMID- 16358896 TI - [Diagnostic difficulties in a patient with middle ear tumor]. AB - Authors present the case of a 57-years old female patient with middle ear tumour, with the history of middle ear operations and reoperations due to chronic inflammatory process. Difficulties in diagnostic procedure related to the kind and size of tumour and to the destruction of surrounding tissues are described as well as operative difficulties connected with the extent and type of surgical intervention. The newest diagnostic methods show with great accuracy the vascular supply of middle ear tumour and the directions of its dissemination within temporal bone. The presence of vast vascularization of middle ear tumour suggests the diagnosis of chemodectoma. PMID- 16358897 TI - [Nasal polyps in a chimpanzee]. AB - The work presents a case of occurrence of nasal polyps of a chimpanzee. The world wide literature holds the view that this pathology does only occur among the human race. Only in two cases the polyps has been found among chimpanzee. PMID- 16358898 TI - [The rare case of sinonasal osteosarcoma in a 22-year-old patient (case report)]. AB - Sarcomas of head and neck are very rare and comprise about 10 to 15% of all malignant nonepithelial neoplasms. A case of 22-year old patient with sinonasal osteosarcoma was presented. The authors stressed the significance of CT in the diagnostic procedures. The preoperative symptoms, histological character of the tumor and postoperative status were shown and discussed. Patient was treated by combined method: the surgical excision and radio-chemotherapy. The 24 months follow-up period was free of symptoms. PMID- 16358899 TI - [Sialolithiasis of the parotid gland--case report]. AB - In this study the very rare case of a parotid gland sialolithiasis was presented. The authors described opinions concerned with symptomatology, localization, diagnostics and methods of treatment in parotid sialolithiasis. The 12 months follow-up period was free of symptoms. PMID- 16358900 TI - [Oncocytic cyst of the larynx--case report]. AB - We report a case of oncocytic cyst of the larynx. It is a rare pathology, usually of a cystic character. Laryngeal lesions have been been described in the literature under a variety names: oncocytic cyst, cystis oncocytica, oncocytic papillary cystadenoma. Detected accidentally, sometimes symptoms are hoarseness or gentle disorders of breathing. It is occurring chiefly at persons after 60 years. The histopathology of these lesions is characterized by proliferation of oncocytic cells. Complete remove this pathology is the treatment of choice, although incomplete excision may be associated with recurrence. Follow up is recommended. PMID- 16358901 TI - [Sarcoidosis limited to the lymph nodes of the neck]. AB - A case of isolated neck sarcoidosis is presented. Isolated sarcoidosis confined to the neck lymph node is rare. Sarcoidosis is a systematic disease usually involving pulmonary findings. Extrapulmonary involvement including the liver, eyes, lymph nodes, central nervous system, muscles and bones is common. This report documents a very rare case of isolated sarcoidosis of the neck, which is very rare diagnosed by otolaryngologists. PMID- 16358902 TI - [Olfactory neuroblastoma in 55 year old woman]. AB - In this article the authors describe a rare case of olfactory neuroblastoma in a 55-year-old woman surgically treated because of nasal polyposis. Three years earlier she had been operated for nasal polyposis on the same side in another ENT department. Unfortunately these lesions may had not been evaluated histologically. One of the surgically removed polyps was atypical. In histopathological examination it was confirmed to be olfactory neuroblastoma. CT scan revealed abnormal thickness of the mucosa in the upper part of the nose and ethmoidal sinuses on the right side. The tumor was classified as stage B in Kadish classification of olfactory neuroblastoma. The histopathologic diagnosis and CT scan made after the surgery resulted in necessity of further oncological treatment. The olfactory neuroblastoma is rare, difficult to diagnose, malignant, slowly growing tumor arising from the olfactory epithelium in the upper nasal cavity. The treatment includes surgery, radiotherapy and chemotherapy. Another objective of this paper is to point out that histopathological examination is crucial in each surgically removed tissue. PMID- 16358903 TI - [Granular cell tumour of the larynx--report of two cases]. AB - Granular cell tumour (Abrikosov tumour) of the larynx is an uncommon laryngeal tumour. We report two cases of this rare type of tumor, which were diagnosed and treated at the Department of Otolaryngology of District Hospital in Zamosc in the period 1999-2005. The tumours were detected in two adult men. In the first case (29 years old patient) the tumour was located next to the right arytenoid. The tumour was removed by microlaryngoscopy. In the second case (55 years old patient) the tumour involved half right vocal fold from the posterior commissure. It was removed by laryngofissurae. Both tumours have not recurred after a 5 years follow-up and are still under observation. PMID- 16358904 TI - [The syndrome of dyskinetic cilia as the cause of chronic sinusitis]. AB - The syndrome of dyskinetic cilia is the cause of chronic inflammatory processes of upper and lower airways. The research studies concerning cilia's structure were held during 50s. About 20 types of cilia's defects were characterized by now, classified as inborn defects. OBJECTIVES: The presentation of the case is because of both the rarity of incidence and diagnostical's problem and treatment. In this case, we observed who re-occurrence of chronic inflammation accompanied by having infections of lower respiratory tract it is necessary to expand diagnosis to find out the reasons of such disease. MATERIALS AND METHODS: A 7 years old girl was admitted to our Clinic. Since couple years she has been under assistance of Specialist Clinic. She was also hospitalized because of aggravation inflammatory state of lower airways. After computers tomography of sinuses she was qualified to surgical treatment. During operation a material to study of cilia immobility was taken. The electron microscope study confirm the diagnosis of the group of dyskinetic cilia. RESULTS: The follow-up examination was held 14 months after operation. During this time the girl did not take any antibiotic (before antibiotherapy was held couple times during the year). Clinical status proved. Better results were also on computer tomography. CONCLUSIONS: In this case, re-occurrence, inflammatory process both upper and lower respiratory tract are necessary to expand the diagnostics to find out the cause of illnesses. The methods of treatments the group of in-movement's cilia based on the symptomatic treatment, but the role of physiotherapy of respiratory system is stressed as the operation of removal of secretion's retention. There is an important role of regular laryngology care and surgical treatment allow partly to control the disease. PMID- 16358905 TI - [Cerebral abscess and ear cancer in the course of chronic otitis media]. AB - Authors are discussing the coexistence of cerebral abscess and the ear cancer in the course of chronic otitis media. They are emphasizing the existing danger of intracranial complications and at the same time the development of cancer with a non-typical course in the middle ear with effusion is more probable. PMID- 16358906 TI - [Retroauricular abscess as a complication of latent otitis media]. AB - In the age of antibiotics intra- and extracranial complications symptoms are latent, and the disease course might be tricky. Despite of low amount of otitis complications its mortality reaches a dozen or so percent. The paper presents two cases of well organised retroauricular abscess in patients with epilepsy and mental impairment. Latent beginning and progress of the disease was underlined. Symptoms, diagnosis process, therapy and treatment results was presented. The authors pay attention to computer tomography importance in diagnostic process. The patients were treated with antibiotics and surgery. Radical ear operation and wide incision and drainage of the abscess were performed, selective therapy with antibiotics was applied. Currently the two patients are under constant laryngologist observation. PMID- 16358907 TI - [Phlegmon neck and face as a complication peritonsillar abscess--case report]. AB - The authors describe a 46-year-old male patient. Inflammatory state of tonsils and peritonsillar abscess caused a serious complication which came to parapharyngeal phlegmon and superficial soft tissue phlegmon face and neck. A wide application of antibiotic gives a rare complication, but mortality reaches a few percent. A basic method is a surgical treatment--a wide cut of tissues in many places, to drainage interfascia space and compound aim intravenous antibiotic therapy. PMID- 16358908 TI - [Rhinosinusitis: classification and definitions]. AB - Rhinosinusitis is a complex disease which is still not fully understood. It has variety of symptoms that may negatively affect quality of life. In order to provide the patients with an efficient diagnosis and effective treatment it is necessary to have the uniform classification and clear definitions of the disease. The objective of this review is to present current state of knowledge of this subject. PMID- 16358909 TI - [Cervical vertigo--etiology, diagnosis and therapy]. AB - The aim of the study was to analyze the reasons and actual methods helpful in diagnostic procedures of cervical vertigo. The vascular, proprioceptive and autonomic etiologies were underlined as responsible for cervical vestibular syndrome. The electrophysiologic methods were introduced together with the visual ones. The treatment was suggested to be directed to the etiology. Rehabilitation, pharmacotherapy, magnetic stimulation and surgical procedures must be taken into consideration in treatment patients with cervical vertigo. PMID- 16358910 TI - [Select neuro-otologic procedures in vertigo cases]. AB - The aim of the study was evaluation of central vestibular disorders in the scope of vestibular ocular reflexes disturbances in three planes: roll, pitch and yaw. The topo-diagnostic value of ocular tilt reaction, vertical nystagmus, rotational and horizontal nystagmus in central lesions was discussed. PMID- 16358911 TI - [Acute sinusitis]. AB - The sinusitis is the term, which describes inflammation process of mucous membrane of these sinuses. Generally inflammation spreads from the nose to the paranasal sinus, so rhinosinusitis is the common name of disease. In classification of sinusitis two aspects are important--duration of signs and symptoms and changes in mucous membrane of sinus. In acute sinusitis symptoms withdraw after treatment, and mucous membrane comes back to the normal state. The most frequent bacteria's responsible for sinusitis are aerobes and among them: Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrrhalis, Streptococcus gr. A, Staphylococcus aureus, bacteria Gram (-) as well as anaerobic bacteria's. The inflammation changes of paranasal sinuses the most often enter on frontal ethmoid sinuses and then in frontal sinus, posterior ethmoid sinuses and the most seldom in sphenoid sinus. The diagnostics of sinusitis consisted of history data, otolaryngological examination, image diagnostics, bacteriological investigation, allergological diagnostics, sometimes alternatively, dentist as well as neurologist's consultations. The treatment of acute sinusitis is conservative. The aim of the treatment is control of inflammation, decrease of tissue oedema, facilitation of outflow of secretions of the nose and paranasal sinuses. We apply the antibiotics, glycocorticosteroids, decongestant drugs and mucolytic drugs. One should remember, that not treated or wrongly treated acute sinusitis can lead to chronic sinusitis or to numerous complications. PMID- 16358912 TI - [The epidemiology and classification of laryngeal carcinoma]. AB - The basic epidemiological data concerning laryngeal carcinoma in Poland were presented, and risk factors in laryngeal carcinoma were reviewed. The premalignant laryngeal lesions, grading of squamous cell carcinoma, and rare malignant tumors of the larynx were discussed. Apart of presentation of actual TNM classification of laryngeal carcinoma, the established criteria of local advancement assessment, and anatomical terms introduced in new TNM system were clarified. PMID- 16358913 TI - [Acute pharyngitis]. AB - The signs and symptoms of acute pharyngitis was described, and principles of diagnostic and therapeutic approach was discussed, concentrating on differentiation between the various forms of virus and bacterial pharyngitis. The economical and clinical importance of rational diagnosis of acute pharyngitis was pointed out, in aspect of antibiotic prescribing limited to acute streptococcal tonsillitis. PMID- 16358914 TI - [Similarity of immunological mechanisms responsible for immunotolerance in healthy subjects and mechanisms induced by specific immunotherapy]. AB - The immonomodulatory mechanisms responsible for allergens tolerance in healthy subjects were described on the basis of the newest medical literature, including own papers and experience. It was detected that the same immunological mechanisms are built by specific immunotherapy. IL-10 and TGF-beta cooperate in the regulatory T-cell response to aeroallergens, food allergens and allergens of bee and yellow jacket venoms in healthy subjects as well as during specific immunotherapy making the tolerance to these allergens. The actual progress in improvement of specific immunotherapy methods is described. PMID- 16358915 TI - [Tuberculosis within the laryngologic organs]. AB - Tuberculosis, considering entry of infection, most frequent concerns the lungs. Rarely we observed extrapulmonary cases of tuberculosis within the bones, joints, urogenital organs and lymph nodes as well as within the head and neck region- larynx, nose and paranasal sinuses, middle ear, pharynx, tonsils, tongue, parotid glands and central nervous system. The diagnosis of head and neck tuberculosis is based on patient's examination as well as on the bacteriologic and histologic study of the biopsy specimen. AIM: The aim of the study was to show our own, very rare cases of head and neck tuberculosis. MATERIAL AND METHODS: We showed our own experience in the diagnosis and treatment of patients suffer from head and neck tuberculosis. 7 cases of head and neck tuberculosis (4 laryngeal, 2 sinonasal and 1 middle ear) were treated from 1986 to 2004 in the II ENT Department Silesian Medical University in Zabrze. In all the cases the diagnosis was based on the histological examination of the tissue specimen. All of the patients were suspected of the neoplastic disease. We assessed good long-term results after the conservative treatment (sometimes completed by surgery). CONCLUSIONS: Tuberculosis of the head and neck region should be taken into consideration during the differential diagnosis of neoplastic diseases. PMID- 16358916 TI - [Natural history of otosclerosis--from tinnitus to deafness]. AB - Authors review the most important data about natural history, signs and symptoms and clinical stages of otosclerosis. Development of otosclerosis is long-lasting and involves several decades of lives. First symptoms already appear in teenagers in about 20% of cases. In advanced cases there are irreversible sensorineural hearing loss. When total deafness occurs cochlear implantation is nowadays the treatment of choice. PMID- 16358917 TI - [Microbiologic diagnosis of acute upper respiratory tract infections]. AB - The main aim of the microbiological diagnosis is to detect the etiologic agent of an infection. In case of bacteria it means to check the susceptibility to antibiotics. It can be difficult to find the etiological agent of respiratory tract infections due to wide range of potential pathogens both viral and bacterial. Culture methods are the most frequently used while in case of atypical and viral pathogens rather serological methods based on specific antibody level checking are recommended. In high standard level laboratories molecular biology methods are more widely used. They allow to detect in short time the existence of typical bacteria like Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, atypical ones like Mycoplasma, Chlamydia, and viral pathogens like influenza virus, parainfluenza virus, adenovirus, RSV and rhinovirus. In fact it is very difficult in Poland to get access to high level laboratories for primary care physicians. This is the reason why the treatment of respiratory tract infections is mainly empirical and based on guidelines developed by experts. PMID- 16358918 TI - [Long-term results of stapes surgery--over 1000 patients]. AB - Authors present audiometric results of 1200 patients with otosclerosis post stapedotomy. Air-conduction, bone-conduction and air-bone gap was measured prior the surgery and 12 months post stapedotomy for frequencies 0.5, 1.0, 2.0 and 3.0 kHz. In the analysed group of 1200 patients a postoperative air-bone gap 0-10 and 11-20 dB was obtained in 93.7% patients. The result of stapedotomy in reference to tinnitus based on the patients' subjective opinion showed that 78.0% patients were either free from tinnitus or tinnitus was subjectively decreased. Authors stress that some anatomical abnormalities in the tympanic cavity might influence on the result of stapes surgery. PMID- 16358919 TI - [Diagnosis and decision making about the management of chronic otitis media]. AB - The paper presents actual state of knowledge in reference to the clinical aspects, diagnosis and indication for surgery in chronic otitis media. The pathologic aspects in the tympanic cavity and in the mastoid should be taken into consideration in the preoperative assessment of the patients. The most common types of chronic non- and cholesteatomatous otitis media and preoperative management are presented. Most of the cases of chronic otitis media require surgery, state of ossicular chain and transmission system determine the functional and anatomical results of the surgery. PMID- 16358920 TI - [Intravenous antibiotics for rhinosinusitis]. AB - The still present problem of choice of a treatment method in chronic sinusitis was introduced in this study. The heterogenous etiology of chronic sinusitis and its treatment consequences were presented. The main current options in the treatment of chronic sinusitis were shown. The functional endoscopic sinus surgery was emphasized as the most important. The different courses of antibiotic therapy: nebulization, oral and intravenous were introduced as a method of choice in exacerbation of chronic process. The intravenous antibiotics were analyzed in details and it was especially based on the bacterial biofilms theory according to the newest literature. The obtaining results were good enough. They were better in children than in adults and in patients with shorter history of sinusitis, however they were not free from complications. Thrombophlebitis, allergic reaction, neutropenia were indicated as most frequent complications that caused the break in the therapy. In the conclusion the intravenous antibiotic therapy was estimated as a useful procedure, but not highly recommended due to a possibility of complications. PMID- 16358921 TI - [Application of implantable electronic prostheses for patients with hearing impairment]. AB - Introduction of implantable electronic prostheses to the clinical practice has created new possibilities of hearing and language communication improvement. Currently implants of various types are used as prostheses implanted in the external ear, temporal bone, middle ear, inner ear and brain stem. Applying appropriate acoustic and electric stimulation they allow us to effectively improve auditory abilities and introduce born deaf patient to the world of sounds or bring back to the world postlingually deafened patient. PMID- 16358922 TI - [Medical management of chronic rhinosinusitis with macrolides]. AB - Long-term, low-dose macrolide therapy is effective in the treatment of chronic airway inflammation. It is believed that macrolide antibiotics produce this benefit through an antiinflammatory effect that is separate from their antibiotic effect. Eosinophils are key mediators in the inflammation seen in chronic rhinosinusitis. The effect of macrolides on eosinophilic inflammation is presented. In vitro studies recently have suggested that macrolides increase eosinophil apoptosis and reduce production of eosinophil chemotactic cytokines and adhesion molecules. In vivo studies have shown a reduction in eosinophil count and activity in asthma and chronic rhinosinusitis. Further in vitro and clinical studies are required to investigate this subject. The authors present preliminary results of clinical trial. PMID- 16358923 TI - [Acute otitis media in children]. AB - Acute otitis media (A.O.M.) occurs mainly in children. The first peak of this disease appears between 6-18 month of life, and the second between 4-7 year of age. The younger age at which the first A.O.M. incidence is noted, the higher probability of its recurrence. In Poland 65% of children up to the 2 year of age suffer from otitis media once, and 30% more than three times. The portal of infection in a.o.m. comprises: (1) Eustachian tube (2) Blood vessels (3) External auditory meatus with perforated tympanic membrane. In ca. 30% of a.o.m. the causative agents are viruses, while in 70% the disease is due to bacteria (Streptococcus pneumoniae in 30-40%, Haemophilus influenzae ca. 20%, Moraxella catarrhalis 10-15%). A.O.M. is diagnosed basing on history, but mainly using otoscopic examination. Regarding different ear anatomy in infants, otoscopic examination may cause many difficulties. In A.O.M. due to possible dehiscence in tegmen tympani or antri may occur meningismus, and dehiscence in the facial nerve canal can occur facial nerve paresis. The treatment of choice is tympanocentesis and administration of amoxycillin in the dose of 40 mg/kg b.w. daily. After recovery, the examination of upper respiratory tract patency should be performed and following that the evaluation of the ear should be continued. PMID- 16358924 TI - The effect of lemon fragrance on simple mental performance and psychophysiological parameters during task performance. AB - The purpose of this study is to effectively utilize fragrance in order to form a comfortable working environment, for which we obtained necessary data. The subjects were 14 female students. We made them do addition work in laboratories with and without lemon fragrance, and investigated their task performances, physiological changes and mood conditions. Our results showed that, although the existence or nonexistence of fragrance did not affect their work efficiency, it was revealed that fragrance mitigated exhaustion and maintained vigor. PMID- 16358925 TI - [A rare case of primary aldosteronism caused by bilateral functioning adrenocortical adenomas with renal cell carcinoma]. AB - We report a rare case of bilateral primary aldosteronism with renal cell carcinoma. A 55-year-old woman developed symptoms of hypertension and hypokalemia. The cause of these symptoms was suspected to be primary aldosteronism, based on the high levels of plasma aldosterone concentrations (PAC) and low levels of plasma rennin activity (PRA), resulting in a high PAC/PRA ratio. Abdominal CT and MRI revealed tumor masses in both adrenal glands, and a large left renal mass. The preoperative diagnosis was primary aldosteronism due to bilateral functioning adrenocortical adenomas and left renal cell carcinoma. The patient underwent left radical nephrectomy and right partial adrenalectomy. The Pathological diagnosis was left renal cell carcinoma and bilateral functioning adrenocortical adenomas. Primary aldosteronism due to bilateral functioning adrenocortical adenomas is relatively rare and its complication with renal cell carcinoma is an extremely rare case. PMID- 16358926 TI - [Examination of physical and psychological health conditions and the influence factors of home helpers]. AB - The primary aim of this study was to examine the physical and psychological health conditions and the influence factors of home helpers. The secondary aim was to suggest a stress management system for home helpers. Self-report questionnaires were distributed to 1147 home helpers who were with home health care agencies in Kitakyushu-city. Responses from 979 home helpers were received, yielding a response rate of 85.5%. A total of 967 home helpers (excluding 12 male home helpers) were used for analysis. The Japanese version of the General Health Questionnaire 28 (GHQ-28) was used to measure the physical and psychological health conditions of the home helpers. In addition, as an influence factor, stress coping was measured by the Lazarus-type Stress Coping Inventory (SCI). Home helpers also filled out the Ego Aptitude Scale (EAS) as a personality measure. Furthermore, the subjects were asked questions about individual background factors, including age, marital status, working hours, years working as a home helper, type of home helper, job satisfaction, job continuation and awareness of stress. The data were analyzed using the Statistical Package for the Social Sciences (SPSS 11.5 J). The groups of full-time or part-time home helpers showed GHQ-28 total scores of 6.86 +/- 5.48 and 5.21 +/- 4.97 (Mean +/- SD): part time home helpers had significantly higher scores than full-time home helpers. The GHQ-28 measure indicated that about 20% of home helpers had physical and psychological symptoms, and about 4% had mid-level depressive symptoms. About half of the subjects were aware of stress. There were significant negative correlations between GHQ-28 total scores and age. Also, there was a statistically significant relation between GHQ-28 and job satisfaction, awareness of stress, type of stress coping and individual personality. Furthermore, there was a statistically significant relation between GHQ-28 and the confrontive coping type, positive reappraisal of SCI, and critical attitude of Ego Aptitude categories. Findings suggested that for the management for home helper work stress, three points are required in the stress management of home helpers: (1) "Work Load", such as organization characteristics, job characteristics, work conditions; (2) "Influence Factors", such as personal characteristics (stress coping type and personality), social support (colleagues/supervisor), and job satisfaction are important; and (3) Enforcement of case manage- PMID- 16358927 TI - [The effects of an exercise program for the female elderly in the community]. AB - In Mizumaki-cho, Fukuoka, a "School of muscle-building for the elderly" was held by the self-help group "MIZUMAKI Iki-iki senior club" once a week from Sep. 2002 to Mar. 2004. This school program included stretching, aerobics and resistance exercises. In this study, we clarified the effect of exercise for the 38 elderly females in this school. Their average age was 65.8 years old. Blood pressure, BMI, body %Fat, VO2 max, grasping power, time of standing on one leg with eyes open, anteflexion while sitting, time of walking for 10 m and time of standing up from sitting on a chair were measured before starting school, after 6 months, 12 months and 18 months. Every measured value except BMI and VO2 max was improved, and these improvements continued for 18 months. Especially, it was shown that time of standing on one leg with eyes open, time of standing up from sitting on a chair and time of walking for 10 m improved by 121.6%, 112.7%, 30.4%, respectively, compared with each measured value at the start. These improvements might produce the effect of fall prevention for the elderly. From the results of this study, it was clarified that regular exercise builds up the muscles of the female elderly, and continuation of exercise also extends the effect of exercise for the female elderly. PMID- 16358928 TI - [Immune responses and immunosuppressive mechanisms in tumor-bearing hosts]. AB - In the 1980s, it was demonstrated that human tumors had tumor antigens, and immune responses against human tumor antigens were induced. These days, various new immunotherapies are attempted to enhance immune responses against tumors and prevent tumor growth and progression. Nonetheless, it is difficult to reject tumors by only immune responses. One of the reasons is an immunosuppressive state in tumor-bearing hosts. It is considered that an immunosuppressive state is induced by various factors from the tumor side or host side, but the precise mechanism is not clear. When we consider the effective immune responses against tumors, we need to make clear and improve the immunosuppressive state in the tumor-bearing hosts. PMID- 16358929 TI - [Out-patient treatment for cancer patients at cancer chemotherapy centers in university hospitals]. AB - At present, one-third of people die of cancer and the number is still increasing in Japan. A safe and effective treatment system is critically required. Recently, the discovery of new drugs and the development of medical oncology promotes out patient treatment for cancer patients. Out-patient treatment in the cancer chemotherapy center of the University of Occupational and Environmental Health Hospital has been started, and many chemotherapy regimens were verified in this center. Not only different organ-specific chemotherapies but also summarizing the oncology team are necessary for performance of the mission. We describe in this review the characteristics of the cancer chemotherapy center. PMID- 16358930 TI - [Long working hours and cardiovascular diseases: a systematic review]. AB - Three years have passed since the countermeasures against the adverse health effects of overwork started in Japan, and fruitful outcomes have been expected. In the current study, a systematic review of articles was performed regarding the association of long working hours (LWH) with cardiovascular diseases (CVD) to obtain recent evidence of their association. An electronic database search was conducted using PubMed among English-written original articles published until December, 2004. A total of twelve articles were found conforming to the study's inclusion criteria, but evidence supporting the association of LWH and CVD was not detected. However, some distinctive studies related to the prevention of CVD were found relating to the concept of sensitive psychosocial factors such as vital exhaustion, and to the statistical modeling of occupational factors and biological indicators with the interaction term of psychosocial factors. Further studies will be needed to clarify the association of LWH and CVD. PMID- 16358931 TI - [Clinical application of ibudilast for elder patients with chronic subdural hematoma]. AB - Ibudilast, an antagonist of platelet-activating factor receptors, was administered to patients with chronic subdural hematoma (CSDH) to assess its effectiveness in preventing recurrence. The remaining volumes of subdural hematomas on brain computed tomography were measured approximately 1-2 months after using ibudilast. The hematomas were significantly smaller and there was no recurrence. Ibudilast administration may be useful in the prevention of recurrence of CSDH. PMID- 16358932 TI - [A study of various causes of influence on inter-child emotions--from a survey conducted on postpartum mothers]. AB - This research was conducted as a self-reporting anonymous survey of 185 postpartum mothers in hospitals in order to clarify the causes of influence on the inter-child emotions of mothers, who are usually the main fosterers of the child. SPSS 12.0 J for Windows was used for statistical analysis, and the relationship between points of inter-child emotion and each questioned item was analyzed with Mann-Whitney U test and Kruskal-Wallis test. As a result, the following causes of influence on inter-child emotions were found: the existence of a key person, having a concrete birth plan during pregnancy, that it was a desired pregnancy, and that the mother was satisfied with her delivery. From this result, the following supports are thought to be important to realize: the importance of puberty education or contraception counseling, the establishment of a support system for mothers, and consistent support during pregnancy to postpartum at medial institutions where parturition is carried out. PMID- 16358933 TI - Looking at lessons on quality from the Medicare pay-for-performance hospital demonstration. AB - In 2003, the new CMS/Premier Hospital Quality Incentive Demonstration Project--a new type of pay-for-performance pilot project--was set up to financially reward hospitals that improved their care in five clinical areas. The top government players announcing the demo may have left for other endeavors, but 268 hospitals affiliated with Premier are participating in the initiative. With preliminary results in from the first year and the first payments due to be sent out this fall, The Quality Letter for Healthcare Leaders looks at what participating hospitals have accomplished in this pilot program with that financial carrot. PMID- 16358934 TI - NQF looks to improve informed consent for individuals with limited health literacy. PMID- 16358935 TI - Consumers increase website use to compare hospitals. PMID- 16358936 TI - The gastrointestinal cholecystokinin receptors in health and diseases. PMID- 16358937 TI - Surgical revascularization and perioperative management in patients with non-ST elevation acute coronary syndromes. AB - PURPOSE: The management and surgical revascularization treatment of patients with acute coronary syndromes (ACS) have undergone great evolution over the past decade. The objective of the present study was therefore to analyze the outcome and predictors of survival in patients unresponsive to maximal non-surgical treatment referred to emergency coronary artery bypass grafting (CABG) with ACS. MATERIAL AND METHODS: Between October 1999 and September 2004, a total of 3571 CABG patients underwent primary isolated CABG at our institution. Out of these, non-ACS (N-ACS) was present in 3124 patients (group 1), 386 patients (group 2) had non-ST-elevation ACS (NSTE-ACS), whereas 61 patients (group 3) had ST elevation ACS (STE-ACS). Clinical data, in-hospital morbidity and mortality were prospectively recorded and studied retrospectively in the groups. RESULTS: Left main stem stenosis was observed in 25%, 32%, and 41%, respectively (P<0.02). Previous myocardial infarction was found in 33%, 43%, and 73% (P<0.001). Overall in-hospital mortality was 1.5% in group 1, 4.2% in group 2, and 13.0% in group 3 (P< 0.001). Logistic regression and receiver operating characteristic analyses identified cTnI as the strongest preoperative predictor significantly related to in-hospital mortality. A preoperative cTnI level above 1.5 ng/ml was the best single predictor for in-hospital mortality amongst patients with ACS. CONCLUSIONS: The present study clearly demonstrates a significant difference of in-hospital morbidity and mortality between patients with ACS undergoing CABG. A more precise patient's risk stratification on admission and improvements in the perioperative management with adjunctive pharmacological therapies and the use of intraaortic balloon counter pulsation may improve patients' outcome. PMID- 16358938 TI - Surgical treatment of congestive heart failure in coronary artery disease. AB - Heart failure (HF) is a pathophysiological condition, when the heart can not provide adequate blood flow to the body organs. The main cause of HF is now ischemic heart disease (IHD), and the number of patients with HF in aging society is growing. HF is becoming the leading cause of death. Medical therapy does not provide satisfactory results in respect of symptoms and survival (5 year survival 28-40%). Therefore there is a trend towards early invasive methods of IHD treatment: percutaneous or surgical revascularisation and surgical reconstruction of myocardial damage. Most common surgical procedure in IHD is coronary artery bypass grafting (CABG). This treatment is safe and effective in patients with normal ventricular function (operative mortality 0.5%, 5 year survival >92%). Results in patients with impaired left ventricular (LV) function are better than conservative therapy, but still not satisfactory (operative mortality 8.4%, 5 year survival 65%). The modern surgical concept for improvement of ventricular function is left ventricular (LV) shape and volume restoration (SVR) accompanied by CABG. In cases of severe damage of myocardium resulting in left ventricular aneurysm or akinesia, SVR improves LV function and prevents further LV remodeling. At present it is under investigation whether SVR is of benefit for moderate-sized ventricles and NYHA class II symptoms. In case of ischemic mitral insufficiency mitral valve repair is a method of choice. The results of combined procedures in Heart Failure group (CABG + MV reconstruction or SVR) are better than CABG alone. Other surgical alternatives for HF treatment are: heart transplantation, ventricular assist devices (VAD), dynamic cardiomyoplasty, constrictive devices and cellular transplantation therapy. Heart transplantation is reserved for younger patients with less comorbidities. Shortage of donor organs and poor long-term results remains a main problem of such a treatment. VAD at present is still very expensive, and serves particularly as a "bridge to heart transplantation" or "bridge to recovery" rather than destination therapy. Despite of all achievements in medical or invasive HF treatment further basic and clinical works as well as new organization systems are necessary to find optimal strategies to reduce cost of care, improve quality of life and survival. PMID- 16358939 TI - Impact of renal dysfunction as a cardiovascular risk factor. AB - This review summarizes the available evidence concerning the relationship between renal dysfunction and cardiovascular risk in non-diabetic patients. Based on numerous studies, there remains no more doubt today, that even minor renal dysfunction, as reflected by microalbuminuria and/or decreased estimated glomerular filtration rate, causes a dramatic increase in cardiovascular risk. Renal dysfunction as a novel risk indicator should be incorporated into currently used algorithms to assess risk factor profile, not to the least because evaluation of renal function helps to select the most appropriate strategy to reduce the cardiovascular risk. PMID- 16358940 TI - Helicobacter pylori infection and gastric MALT lymphoma. AB - Helicobacter pylori infection is implicated in the development of two different gastric cancers: gastric adenocarcinoma and gastric MALT lymphoma. The association with the gastric MALT lymphoma is strong and causal. It is currently the only cancer which can be treated by a simple antibiotic treatment. However, the evolution of an H. pylori infection towards lymphoma is exceptional. Host susceptibility factors and environmental factors predisposing a patient to lymphoma have not yet been determined. The bacterial factors are currently being identified. PMID- 16358941 TI - Somatostatin-receptor mediated diagnosis and treatment in gastrointestinal neuroendocrine tumours (GEP-NET's). PMID- 16358942 TI - Important clues to the diagnosis of pancreatic cancer. AB - The incidence of pancreatic carcinoma is recently increasing but the prognosis remains extremely poor. Widespread awareness of important clues to the diagnosis is particularly important to improve the prognosis. Dilatation of the main pancreatic duct on ultrasonograms and/or CT scans, hyperamylasemia incidentally found during routine blood examinations, and recent onset diabetes mellitus must lead to thorough imaging studies of the pancreas. Death from pancreatic carcinoma occurs in 0.2-1.9% of all diabetic patients, being more than 300 times frequent compared to general population. Diabetes may be the only clinical sign of pancreatic carcinoma in some patients. In our recent study, of 163 diabetic patients selected by several criteria who underwent ERCP screening, 12 patients (7.4%) proved to have pancreatic carcinoma. The prevalence of pancreatic carcinoma was more frequent in those with a recent onset (<3 years) of diabetes (13.7% (8/58)) than in those with a longer history (>3 years, 3.8% (4/105)). Furthermore, intraductal papillary mucinous neoplasm (IPMN) is reported to be associated with pancreatic carcinoma. Concomitant carcinoma was found in 9 of our series of 94 patients (9.5%) who underwent surgical resection of branch duct IPMN. Of particular interest is the fact that two of the 9 patients had carcinoma in situ that could be diagnosed only by cytology of the pancreatic juice. IPMN may be the only clue to the early diagnosis of pancreatic carcinoma presenting with no clinical symptoms or abnormalities on imaging studies. PMID- 16358943 TI - Hereditary pancreatitis and secondary screening for early pancreatic cancer. AB - Hereditary pancreatitis is an autosomal dominant disease with incomplete penetrance (80%), accounting for approximately 1% of all cases of pancreatitis. It is characterized by the onset of recurrent attacks of acute pancreatitis in childhood and frequent progression to chronic pancreatitis. Whitcomb et al. identified the cationic trypsinogen gene (PRSS1) on chromosome 7q35 as the site of the mutation that causes hereditary pancreatitis. The European registry of hereditary pancreatitis and familial pancreatic cancer (EUROPAC) aims to identify and make provisions for those affected by hereditary pancreatitis and familial pancreatic cancer. The most common mutations in hereditary pancreatitis are R122H, N29I and A16V but many families have been described with clinically defined hereditary pancreatitis where there is no PRSS1 mutation. It is known that the cumulative lifetime risk (to age 70 years) of pancreatic cancer is 40% in individuals with hereditary pancreatitis. This subset of individuals form an ideal group for the development of a screening programme aimed at detecting pancreatic cancer at an early stage in an attempt to improve the presently poor long-term survival. Current screening strategies involve multimodality imaging (computed tomography, endoluminal ultrasound) and endoscopic retrograde cholangiopancreatography for pancreatic juice collection followed by molecular analysis of the DNA extracted from the juice. The potential benefit of screening (curative resection) must be balanced against the associated morbidity and mortality of surgery. Philosophically, the individual's best interest must be sought in light of the latest advances in medicine and science following discussions with a multidisciplinary team in specialist pancreatic centres. PMID- 16358944 TI - Extent of lymphadenectomy in the resection of pancreatic cancer. Analysis of the existing evidence. AB - Pancreaticoduodenectomy is considered the standard procedure for the surgical treatment of the pancreatic head cancer. However, the extent of lymph node clearance associated to the procedure is still largely debated. Arguments in favour of an extended lymphadenectomy are the regular progression of lymph node invasion, without skip metastases, and the removal of the extrapancreatic neural plexus that is invaded in 52-72% of patients. Arguments against the extended lymphadenectomy are the failure of extended lymphadenectomy to improve survival in other cancers, and the severe diarrhoea that follows the skeletonisation of the superior mesenteric artery. After Ishikawa's paper, several retrospective studies supported a longer survival after an extended than after a standard lymphadenectomy, but as much retrospective studies failed to demonstrate any difference. Only three prospective randomised controlled trials have been performed so far. Unfortunately all are underpowered, and the substantial differences in the surgical procedures, in the adjuvant treatment, and in the length of follow-up make the comparison impossible. Only one study reports a significantly longer survival for lymph node positive patients who underwent an extended lymphadenectomy, but adjuvant treatment was not performed. Furthermore, the difference was of minimal clinical impact. At least two adequately powered prospective Randomised Controlled Trials including a true extended lymphadenectomy, and a standardised adjuvant treatment, would be required to answer the question. Unfortunately, we have not yet a standardised adjuvant (or neoadjuvant) treatment, and we do not know the impact of such treatment on the expected statistical difference in the survival after a standard or extended lymphadenectomy. The lot of work required to perform such trials probably doesn't worth the expected results. PMID- 16358945 TI - Application of specific cytologic, cytogenetic and molecular-cytogenetic techniques for the characterization of solid tumors. AB - Investigations of solid tumors have shown that a very specific characterization of aberrant tissues can best be performed using a combination of cytologic, cytogenetic and molecular-cytogenetic methods. Thus, cytological analyses may serve to examine various features of tumors cultivated in vitro, e.g. growth peculiarities, cell morphology, specific details of cell division and mitotic rates, and anomalies of the spindle apparatus. Besides, chromosomal diagnostics characterizing non-specific aberrations focuses on the pathological karyotype and its evolution and heterogeneity, as well as on the development of secondary chromosomal aberrations. In the field of molecular-cytogenetic diagnostics we emphasize particularly the combination of metaphase and interphase analyses and the investigation of specific structural aberrations by fluorescence in situ hybridization (FISH). In contrast to the method of comparative genomic hybridization (CGH), the spectrum of applications for both methods is discussed. The findings described in this paper were obtained primarily from the analysis of 68 tumors of the urogenital tract (20 kidney tumors, 33 bladder tumors, 15 testis tumors). PMID- 16358946 TI - Progresses in the medical treatment of advanced colorectal cancer. AB - In the last 2 decades, major progresses have been made in the management of patients with advanced colorectal cancer (ACC). The modulation of 5-fluorouracil (5-FU) by folinic acid (LV), followed by the introduction of irinotecan and oxaliplatin have significantly improved the outcome of these patients. New strategies consist of oral fluoropyrimidines, and of targeted agents to inhibit cancer signalisation. PMID- 16358947 TI - New strategy for acute necrotizing pancreatitis: Continuous Regional Arterial Infusion (CRAI) therapy. AB - Acute pancreatitis is an autodigestive disease, of which protease inhibition has been the focus of experimental and clinical research. Different from Europe and the United States, protease inhibitors are often applied in the treatment of acute pancreatitis in Japan. However, in clinical settings, the effect of protease inhibitors on acute pancreatitis is still controversial. Continuous Regional Arterial Infusion (CRAI) of protease inhibitors and antibiotics therapy were developed in Japan and it has been demonstrated that CRAI therapy has beneficial effects on severe acute necrotizing pancreatitis. In the Japanese clinical guidelines for the treatment of acute pancreatitis, published in 2003, CRAI therapy is still classified as a special therapy. However, a Randomized Controlled Trial for CRAI therapy has started and CRAI therapy is expected to become a new standard therapy for severe acute pancreatitis. CRAI therapy is aimed at preventing the progression of pancreatic inflammation and pancreatic infection. CRAI therapy can decrease the mortality rate and the frequency of pancreatic infection in severe acute pancreatitis, but it should be started as soon as possible after the onset of acute pancreatitis. PMID- 16358948 TI - Anti-cytokine strategies in acute pancreatitis: pathophysiological insights and clinical implications. AB - The clinical presentation of acute pancreatitis varies significantly from mild self-limiting discomfort to a severe life-threatening condition. Once the disease process is initiated, the severity of the disease is largely determined by a complex network of activated inflammatory mediators such as cytokines, proteolytic enzymes, reactive oxygen species, and many more which render the local injury to a systemic disease with multiple organ dysfunction, sepsis, and considerable mortality. Remarkable progress in diagnostic modalities, intensive care technologies, and organ preserving surgical techniques have decreased mortality of severe acute pancreatitis during the past decades. However, the treatment of acute pancreatitis still remains largely supportive and no specific approach exists to prevent evolving complications. A large body of clinical and experimental evidence suggests that cytokines are key factors in the pathomechanism of local and systemic complications of acute pancreatitis. Targeting cytokine activity as therapeutic approach to acute pancreatitis is a challenging concept and the results of modulating activation of TNF-alpha, IL 1beta, IL-2, IL-10, PAF and various chemokines has indeed been promising in the experimental setting even if tested under therapeutic conditions. However, experience from a limited number of clinical trials on anti cytokine strategies in acute pancreatitis has remarkably emphasized that translating successful experimental observations into reproducible clinical associations seems to be difficult. PMID- 16358949 TI - Antibiotic treatment in acute pancreatitis. AB - Severe acute pancreatitis is characterized by a poor prognosis with local and systemic complications, high morbidity and mortality. From the morphological standpoint, almost all patients suffering from severe forms of acute pancreatitis present various degree of pancreatic necrosis. In these patients the occurrence of infection of pancreatic necrosis certainly represents a very important prognostic factor as it has worldwide accepted as the leading cause of death. In addition, the discovery of an infected necrosis represents a crucial point in the treatment of these patients as it is the only clear-cut shift from medical to surgical treatment in necrotizing pancreatitis. Over the last years, earlier and more precise identification of pancreatic necrosis together with availability of new classes of antibiotics with documented activity against the most commonly involved bacteria and able to reach in therapeutic concentration the pancreatic necrosis give us the opportunity to perform some important controlled clinical trials on antibiotic prophylaxis in necrotizing acute pancreatitis. The great majority of these studies showed the usefulness of a prophylactic regimen (using antibiotics such as fluoroquinolones and carbapenems) in terms of reduction of pancreatic and extrapancreatic infections in comparison with untreated controls. Nevertheless, some questions on this topic still present controversial aspects such as the antibiotic of choice, the duration of treatment, the possible opportunistic infections with fungi and/or resistant strains. Antibiotics may prove very useful in patients with documented infected necrosis and high anaesthesiological risk unfit for surgical debridement and drainage; some initial experiences show the possibility that antibiotic treatment may be curative without surgery in these selected cases. PMID- 16358950 TI - Diabetic nephropathy and cardiovascular diseases. AB - Diabetic nephropathy is diagnosed either when persistent increase of urinary albumin excretion rate (UAER) above 30 mg/24h in a patient with diabetes was discovered (early or incipient nephropathy) or when UAER values are persistently elevated above 300 mg/24h (overt or clinical nephropathy). In both situations the additional criteria of presence of diabetic retinopathy and the absence of the evidence of other kidney or renal tract disease should be fulfilled. It was found that the excess of cardiovascular events and mortality occurs already in diabetic patients with persistent microalbuminuria, but is particularly evident in macroalbuminuric diabetic patients and results not only from end-stage renal failure (ESRF) but rather from cardiovascular disease (CVD), the latter mainly in type 2 diabetic patients. Several traditional risk factor for atherosclerosis has been identified in diabetic patients with micro- or macroalbuminuria including elevated blood pressure levels, dyslipidemia and procoagulatory state associated with endothelial dysfunction. Microalbuminuria is currently regarded as a marker of generalized endothelial damage, it reflects transvascular albumin leakage, now recognized as an early event in atherogenesis. Recently the association of microalbuminuria with the marker of chronic inflammation (C-reactive protein) and with increased production of vascular endothelial growth factor (VEGE) was described. Thus, multiple mechanisms are involved in the development and progression of cardiovascular complications both in micro- and macroalbuminuric diabetic patients and all these mechanisms should be regarded as the target for therapeutic intervention. PMID- 16358951 TI - Hemostasis in chronic renal failure. PMID- 16358952 TI - A grand challenge for research: multimodal, multilevel and multiscale systems in medicine and biology. AB - Computational modelling, nano-bioscience and information technology in biology and medicine will play a major role in the interdisciplinary attempts to elucidate structures and functions of living systems. Developing tools capable to integrate the new advances and make benefit of them is crucial: accumulation of data and knowledge base with only storage and retrieval capabilities will have a poor impact if they are not made "active" or "operational". This is where models will play a central role in offering, not only sound ways for representation or simulation, but also the appropriate frames to put the players in the right place, with intra- and inter-level coupling and multisource handling. This paper advocated that sequential observations of multiple and complex mechanisms will be of limited interest to understand the inter-relations that are occurring at the same time, and therefore, that designing multimodal, multilevel and multiscale experiments, matched with these models, are of major importance. PMID- 16358953 TI - New approaches to health promotion and informatics education using Internet in the Czech Republic. AB - The paper describes nowadays information technology skills in the Czech Republic. It focuses on informatics education using Internet, ECDL concept and the links between computer literacy among health care professionals and quality of health care. Everyone understands that the main source of wealth of any nation is information management and the efficient transformation of information into knowledge. There appear completely new decisive factors for the economics of the near future based on circulation and exchange information. It is clear that modern health care cannot be built without information and communication technologies. We discuss several approaches how to contribute to some topics of information society in health care, namely the role of electronic health record, structured information, extraction of information from free medical texts and sharing knowledge stored in medical guidelines. PMID- 16358954 TI - Peroxisome proliferator-activated receptor gamma ligand prevents the development of chronic pancreatitis through modulating NF-kappaB-dependent proinflammatory cytokine production and pancreatic stellate cell activation. AB - PURPOSE: Thiazolidinedione derivatives (TZDs) are known to be ligands of peroxisome proliferator-activated receptor gamma (PPARgamma). In this study, we investigated the effect of a TZD, troglitazone, on inflammation and fibrogenesis in the pancreas of an experimental model of chronic pancreatitis. MATERIAL AND METHODS: Male WBN/Kob rats with spontaneous chronic pancreatitis were fed rat chow containing 0.2% troglitazone from 1 to 4 months of age. Immunohistochemical studies of rat pancreas were carried out with monoclonal mouse antibody against human alpha-smooth muscle actin (alpha-SMA) or rabbit polyclonal antibody against collagen type I, collagen type III, or fibronectin. Cytokine production was measured by enzyme-linked immunosorbent assay. The inhibitory action of troglitazone on nuclear factor-kappaB (NF-kappaB) binding activity in activated macrophages was also investigated. RESULTS: Long-term administration of troglitazone reduced inflammatory cell infiltration and fibrosis in the pancreas of WBN/Kob rats, and expression of alpha-SMA, procollagen I, III, and fibronectin was significantly reduced by troglitazone. The increase in TNF-alpha production by activated macrophages was significantly decreased by troglitazone. Peritoneal macrophages isolated from WBN/Kob rats produced a large amount of TNF-alpha, whereas those from troglitazone-treated WBN/Kob rats produced only a marginal amount of TNF-alpha. Lipopolysaccharide-induced NF-kappaB binding activity in peritoneal macrophages was also significantly reduced by troglitazone. CONCLUSIONS: Troglitazone prevented the progression of chronic pancreatitis via inhibition of ECM synthesis and proinflammatory cytokine production mediated by the inhibition of NF-kappaB activity. PMID- 16358955 TI - Single Nucleotide Polymorphisms in exon 3 of the adiponectin gene in subjects with type 2 diabetes mellitus. AB - PURPOSE: Adiponectin (APM1)--a newly discovered adipocytokine secreted by fat tissue--was recently suggested to play a role in the genetic predisposition to type 2 diabetes, obesity and insulin resistance. Adiponectin gene is localized on chromosome 3q27 within the region which was identified as susceptibility locus for type 2 diabetes and metabolic syndrome. Till now genetic associations of two SNP in exon 2 (+45T/G) and intron 2 (+276G/T) of adiponectin gene with type 2 diabetes and adiponectin level were reported in Japanese population and with insulin resistance in some Caucasian populations (Italy, Germany). Moreover, in the proximal promoter region of the APM1 gene: SNP-11426A/G and -11391A/-11377G haplotype predicted the associations with fasting plasma glucose, type 2 diabetes and adiponectin levels. On the other hand the role of mutations in exon 3 of the adiponectin gene is not so well studied. MATERIAL AND METHODS: The aim of our study was the screening for rare mutation in exon 3 of adiponectin gene in the Polish subjects with type 2 diabetes as there is no data available about the frequency and role of these mutations in our population. The study was performed in the group of 187 Polish origin patients with type 2 diabetes (32 female and 155 male, mean age 54.1 +/- 8.6 yrs) and 102 age and sex matched healthy controls. RESULTS: The frequency of adiponectin gene mutations in exon 3 was 3.9%, while in the control group 0.98% and this difference was not statistically significant. We also observed that adiponectin level is significantly lower in patients with c.331 T-->C mutation (Y111H) in comparison to subjects without this mutation (5.0 ug/ml vs 14.4 ug/ml, p=0.0148). CONCLUSIONS: To our knowledge the present study is the first which shows that in Polish populations. PMID- 16358956 TI - Interleukin 18 and sICAM-1 serum levels in families with type 1 diabetes mellitus. AB - It is well known that subjects with type 1 diabetes are at an increased risk of death from coronary heart disease in comparison to non-diabetic age-matched individuals because hyperglycaemia is believed to be a key risk factor for the development of micro- and macrovascular complications. On the other hand there is increasing evidence about the role of inflammatory mediators in the pathogenesis of atherosclerosis and the development of acute coronary syndromes. It has been recently suggested that IL-18 and sICAM-1 have a strong predictive value for cardiovascular diseases and deaths in patients with coronary artery disease and/or in apparently healthy men. The aim of our study was to estimate the serum levels of IL-18 and sICAM-1 in subjects with type 1 diabetes and their relatives, who share HLA diabetic susceptibility genes (with or without pancreatic autoantibodies), but still without glucose level disturbances, as an evaluation of the possible role of genetic predisposition to the presence of IL-18 in diabetic families. The study was carried out in 35 type 1 diabetic subjects, their 101 healthy first-degree relatives: 36 siblings and 65 parents and the control group consisted of 31 healthy volunteers. We have found increased IL-18 and sICAM-1 levels in subjects with type 1 diabetes and their first degree relatives, who share diabetic HLA haplotypes: DRB1*03/DRB1*04 or DRB1*03/*04/DQB1*02 independently of their autoimmune status. There was a strong positive correlation between IL-18 and sICAM-1 levels in diabetic subjects and their first degree relatives without glucose level disturbances. To our knowledge this is the first study, which suggests that sICAM-1 elevations could be a result of IL-18 overproduction in type 1 diabetic subjects and their first degree relatives. Since in previous studies IL-18 and sICAM-1 were found to be predictors of death in subjects with CHD, one could speculate that high levels of IL-18 observed in subjects with genetic predisposition, but still with normal glucose levels, are an in addition to hyperglycaemia, pathogenic factors responsible for a higher risk of acute coronary events in subjects with diabetes type 1. PMID- 16358957 TI - Long-term outcome of percutaneous transhepatic drainage for benign bile duct stenoses. AB - PURPOSE: The occurrence of benign bile duct stenoses is mostly associated with prior biliary surgery, pancreatic diseases or sclerosing cholangitis. It remains a challenging problem for gastroenterologists and surgeons, especially in case the endoscopic approach is not possible. The exact role of percutaneous transhepatic stenting for these patients has not been clearly defined yet. MATERIAL AND METHODS: 36 patients with symptomatic benign bile duct stenoses or strictures after surgery underwent percutaneous transhepatic stenting and were studied prospectively. We were particularly interested in how many patients would achieve resolution of the stricture and tolerate removal of the drainage in the long-run. RESULTS: The primary success rate of percutaneous transhepatic biliary drainage (PTBD) was 92% (33/36 patients). All patients presented improvement of jaundice and cholestasis. Relief of the stricture and clinical improvement was achieved in 72% (26/36) of patients after a median stenting time of 14.5 (6-34) months. 5.5% (2/36) required further stenting due to a persistent stricture. A clinical recovery without radiological stricture regression after stenting demonstrated 22% (8/36) of patients. Long-term failures were noted in 27% (10/36) of patients after a median follow-up of 48 months. CONCLUSIONS: Percutaneous transhepatic stenting of symptomatic benign biliary strictures is safe and highly effective in achieving adequate internal bile drainage. There seems to be a therapeutic benefit not only for short-term interventional treatment but also as a sufficient long-term therapeutic alternative to surgery with tolerable complication rates. PMID- 16358958 TI - Trends in the incidence of the free wall cardiac rupture in acute myocardial infarction. observational study: experience of a single center. AB - PURPOSE: Free wall cardiac rupture (CR) is one of the most common cause of in hospital death in acute myocardial infarction (AMI). The early diagnosis of CR and selection of the patients predisposed to CR become an important clinical tool. AIM: assessing the occurrence of CR in patients with AMI, to determine the factors which could help to identify the patients threatened with CR. MATERIAL AND METHODS: 2320 consecutive patients with AMI. CR was proved by autopsy or by echocardiography performed during cardio-pulmonary resuscitation (CPR). RESULTS: In-hospital mortality was 11% (254 patients). 50 patients (2%) died from CR. CR was the cause of 20% of total in-hospital death. Patients with CR were older than survivors (72 vs 60 years, p<0.0001). Women prevailed in CR group: (62% in CR group vs 27% in the survivors, p<0.01). 29% of patients were treated with thrombolytics (Th+). Out of 58 patients from Th (+) group who died, 17 (29.31%) died because of CR. CR occurred in 33 (16.8%) patients out of 196 died in Th (-) group. In the logistic regression analysis only age and sex remained as predictors of CR. 16 patients died from CR during first 24 h from admission (ECR). In 34 patients CR occurred >24 h (LCR). In ECR group were no prevalence of women, while in LCR women constituted 68%. In ECR group all but one patient had no previous history of MI (p=0.06). Frequency of thrombolythic therapy was equal. CONCLUSIONS: Advanced age patients, particularly women with first AMI are at risk of CR. Decision of thrombolytic treatment in this group of patients must be very cautious. PMID- 16358959 TI - Thrombin activatable fibrinolysis inhibitor (TAFI) in stable angina pectoris patients undergoing coronary artery bypass grafting (CABG). AB - PURPOSE: Thrombin activatable fibrinolysis inhibitor (TAFI) seems to be a potential haemostatic risk factor of coronary artery disease (CAD). Taking into account interactions between TAFI and haemostasis, especially during cardiopulmonary bypass, we decided to determine concentration of TAFI and activated TAFI (TAFIa) and other haemostasis markers in CABG patients. MATERIAL AND METHODS: 45 CAD patients (11 women, 34 men) undergoing elective CABG were included in the study. Blood samples were taken before the operation, on the 3rd, 7th day and 3 months after CABG. A value of p<0.05 was considered statistically significant. RESULTS: We found a significant decrease in TAFIa concentration on 3rd postoperative day: 6 microg/ml (0.3-43.2) vs 8.9 microg/ml (0.5-37) before CABG (p<0.05), a significant increase in TAFI concentration on the 7th postoperative day: 127.7% +/- 36.8 vs 112.18% +/- 30.34 of standard plasma concentration before CABG (p<0.05), significant increase in plasmin-antyplasmin (PAP) complexes concentration on 3rd and 7th day, respectively: 645 microg/l (323 1237) vs 406 microg/l (197-1840) before CABG (p<0.001); and 1030 microg/l (640 2149) vs 406 microg/l (197-1840) before CABG (p<0.0001). Before operation we found a significant negative correlation between PAP complexes concentration before CABG and EuroSCORE risk scale value (p<0.01). CONCLUSIONS: In CABG patients, there is a significant increase in fibrinolytic activity due to decrease in TAFIa concentration, with simultaneous increase in PAP complexes. A significant negative correlation between PAP complexes concentration before CABG and EuroSCORE risk scale value stressed a potentially higher operation risk in patients with lower fibrinolytic activity. PMID- 16358960 TI - Concentration of interferon-inducible T cell chemoattractant and monocyte chemotactic protein-1 in serum and cerebrospinal fluid of patients with Lyme borreliosis. AB - PURPOSE: Chronic inflammation in Lyme borreliosis may be sustained by aberrant inflammatory response, characterized by Th1 lymphocyte predominance, which in turn may be determined by chemokines synthesized in inflammatory focus. The aim of the study was to evaluate synthesis of chemokines: interferon-induced T cell chemoattractant (I-TAC--chemoattractant for Th1 lymphocytes), and monocyte chemotactic protein (MCP-1) in Lyme borreliosis. MATERIAL AND METHODS: Study group consisted of 13 patients with erythema migrans, 10 with Lyme arthritis and 6 with neuroborreliosis. Serum, as well as cerebrospinal fluid (CSF) in neuroborreliosis, was obtained before (examination 1) and during (examination 2) antibiotic treatment. Control serum was obtained from 8 healthy volunteers and control csf from 8 patients in whom meningitis and neuroborreliosis was excluded after diagnostic lumbar puncture. The samples were assayed for MCP-1 and I-TAC by ELISA. RESULTS: Serum mean I-TAC concentration in examination 1 was 73.0 pg/ml in erythema migrans, 78.9 pg/ml in Lyme arthritis and 87.3 pg/ml in neuroborreliosis (29.9 pg/ml in controls, difference significant for neuroborreliosis) and did not change significantly in examination 2. MCP-1 serum concentration was significantly increased to 497.5 pg/ml in neuroborreliosis in examination 2. I TAC concentration in csf remained low, while MCP-1 concentration in examination 1 was increased to 589.1 pg/ml, significantly higher than simultaneously in serum. CONCLUSIONS: I-TAC synthesis is increased in Lyme borreliosis and may be a factor favoring predominance of Th1 lymphocyte subset. MCP-1 creates chemotactic gradient towards central nervous system and may contribute to csf pleocytosis in neuroborreliosis. PMID- 16358961 TI - HBV-DNA and sFas, sFasL concentrations in serum of healthy HBsAg carriers. AB - PURPOSE: Increased HBV-DNA concentration is a prognostic factor of disease progression in chronic hepatitis B patients. Moreover, active hepatic inflammation during HBV replication influences apoptosis intensification. The aim of this study was to estimate occurrence of HBV replication among carriers of HBsAg. Furthermore, we analysed the correlation between HBV replication and HBeAg or anti-HBe presence as well as known apoptosis indicators--sFas and sFasL concentration. MATERIAL AND METHODS: The study included 34 HBV infected patients, aged 20-43 yrs defined as HBsAg healthy carriers. HBV-DNA was extracted from patients' serum using two different DNA isolation kits: the QIAamp DNA Mini Kit (QIAGEN Ltd, USA) and the Gene Elute Mammalian Genomic DNA Miniprep Kit (Sigma, USA). HBV-DNA concentration in serum was measured by RT-PCR based on TaqMan Universal Master Mix (Applied Biosystems). The detection limit of this system was as few as 10 HBV-DNA copies/mL of serum. HBV-DNA concentration was calculated from a linear standard curve obtained between 10 and 10(8) DNA copies/reaction. HBeAg and anti-HBe in serum were detected by MEIA method (ABBOTT, Germany). The concentration of sFas and sFasL in serum was-estimated by ELISA method (Bender MedSystems, Austria). RESULTS: HBV active replication was detected in 79% HBsAg carriers. The HBV-DNA levels exceeding 10(5) copies/mL were observed in 64% patients. Among HBsAg carriers presenting HBeAg, HBV replication occurred more often and was more intensify than in HBsAg carriers presenting anti-HBe antibodies. The sFasL occurrence in serum of 56% HBsAg carriers shows an active apoptosis, independent from ALT and AST activity within normal ranges. PMID- 16358962 TI - Morpho-functional comparisons in Helicobacter pylori-associated chronic atrophic gastritis. AB - PURPOSE: To evaluate serum pepsinogen I (PG I) and gastrin-17 (G-17) levels in patients with Helicobacter pylori (H. pylori)-associated chronic atrophic gastritis, with reference to endoscopical Kimura-Takemoto's staging, chromoendoscopical and histological features. MATERIAL AND METHODS: 267 dyspeptic H. pylori-infected patients were examined by chromoendoscopy with biopsy sampling according to the Sydney System and according to Kimura-Takemoto's scale. Simultaneous assessment of serum pepsinogen I (PG I) and gastrin-17 (G-17) levels by enzyme immunoassay was performed. The serologic and morphologic results were compared with correlation analysis. RESULTS: There was strong reverse correlation between the stomach mucosal atrophy (antral part or corpus) and the proper serologic markers (respectively, G-17 or PG I) in H. pylori-associated chronic gastritis when gastric biopsies taken according to the Sydney System were assessed. The use of Kimura-Takemoto's scale has revealed the decrease of serum PG I levels only at 0-2 and 0-3 grades of the corpus mucosa atrophy. Probably, these results reflects the development of functional failure of the stomach corpus mucosa at late stages of atrophy when its compensatory capacity becomes insufficient. There were not any advantages in sampling biopsies for the detecting of intestinal metaplasia (IM) by the Sydney System, or by Kimura Takemoto's scheme. The obvious concordance between histologically proven extent of IM and the number of IM foci detected by chromoendoscopy has been revealed. CONCLUSIONS: The biopsy sampling for the diagnosis of precancerous changes of the stomach mucosa after non-invasive screening of atrophic gastritis (e.g., by means of EIA) should be based preferably on the visual signs acquired via chromoendoscopy than through routine endoscopy, independently of the scheme of examination of stomach mucosa, either according to the Sydney System, or to the Kimura-Takemoto's scale. PMID- 16358963 TI - Gastric juice ammonia and urea concentrations and their relation to gastric mucosa injury in patients maintained on chronic hemodialysis. AB - PURPOSE: This study was undertaken to test the hypothesis that high concentrations of urea in gastric juice would have an influence on Helicobacter pylori infection in patients maintained on chronic hemodialysis (HD). MATERIAL AND METHODS: We investigated 30 patients (17 males, 13 females; mean age 50.8 +/- 2.9 years) with end-stage renal disease (ESRD) undergoing hemodialysis treatment (HD) for at least 6 months, who were compared to 31 patients (16 males, 15 females; mean age 61.3 +/- 2.2 years) with dyspeptic symptoms. Biopsies from the gastric antrum and body were taken for histological investigation. Urea and ammonia were measured in gastric juice, and the severity of gastritis was evaluated according to Sydney criteria. RESULTS: H. pylori infection was found in 19 (63%) HD patients and in 22 (71%) control subjects. Gastric juice urea concentration was significantly higher in HD patients than in controls and H. pylori infection caused a significant decrease in urea concentration in both groups. There was an inverse correlation between urea and ammonia concentration in gastric juice in both groups. Ammonia concentration in both groups was higher in H. pylori infected patients. In H. pylori negative subjects ammonia/urea ratio was lower in HD patients in comparison to controls. Ammonia/urea ratio was raised by H. pylori infection in both groups, and the difference between HD and control groups persisted. H. pylori infection was associated with polymorphonuclear infiltration of gastric mucosa. There was a significant correlation between gastric ammonia and mucosal polymorphonuclear leukocytes infiltration and gastritis score. CONCLUSIONS: Higher urea levels in the gastric juice of chronically hemodialyzed patients do not seem to be a risk factor for infection with Helicobacter pylori. PMID- 16358964 TI - Laser resection of lung parenchyma--a new technical and clinical approach. AB - The introduction of a new 1318 nm wavelength Nd:YAG laser has created new possibilities in lung parenchyma surgery. The potentially curative surgical resection of pulmonary metastases in suitably selected cases had been recognized slowly. Using the new laser technology a greater number of patients can now offered salvage surgery. This paper reviews the history of surgical management of pulmonary metastases, development of new laser technology, conventional and extended indications for pulmonary metastasectomy and use of laser in thoracic surgery. PMID- 16358966 TI - An assessment of telemedicine possibilities in massive casualties situations. AB - The use of existing possibilities of Telemedicine Center of Kaunas University of Medicine allows the live distant consultations from high-level medical specialised centers to rural areas. On July 2004 the Telemedicine Center took part in the RESCUER/MEDCEUR project exercise. A special objective was the use of telemedicine facilities for distant consultations and sorting of victims directly at the event place. Telemedicine Center used appropriate telecommunication devices for joint activities of civil and multinational military services in critical situations such as mass casualty events. There were used ISDN lines and IP radio-connection. On the final and most intensive day of the anti-terrorism drills, the multinational force of medics at the Kairiu Training Range in Lithuania reacted to a large mass casualty event--treating hundreds of victims from a simulated train crash. Using on-line telemedicine facilities from Kaunas Medical University Hospital there were corrected the tactics of giving the first help and sorting of casualties. The most complicated initiated cases of eye trauma, neurosurgical trauma, maxilloface trauma and traumatic amputation of limbs evaluated and selected for emergent evacuation to the third level hospitals. All those cases transported to Kaunas and Vilnius Universities Hospitals by helicopters (200 and 300 km from the event place). The common use of existing military and civil telemedicine infrastructure showed the possibilities of interaction in management, giving the first help and sorting of casualties between military and civil medical services during the rescue operations. PMID- 16358965 TI - Serum endostatin levels in patients with lung carcinoma. AB - The purpose of our study was to evaluate the clinical usefulness of serum endostatin levels during chemotherapy of lung cancer in relation to the histopathological type of the tumor, clinical stage and response to therapy. MATERIALS AND METHODS: Serum concentrations of endostatin were determined in 37 patients (24 with non-small cell lung cancer and 13 with small cell lung cancer), 10 healthy subjects constituted controls. To determine endostatin levels (ELISA), venous blood samples were collected from each patient before treatment and after 4-6 courses of chemotherapy. RESULTS: The serum concentrations of endostatin were found significantly higher in patients in comparison with controls (p=0.003). No statistically significant differences were established between the concentrations of endostatin with regard to such clinical features, as: performance status, clinical stage (III and IV) and histopathological type (non-small cell lung cancer and small cell lung cancer). The concentrations of endostatin did not change after chemotherapy. There was no change of endostatin concentration caused by the response to treatment. CONCLUSIONS: The serum endostatin concentrations were elevated in lung cancer patients. PMID- 16358968 TI - Relationship between insulin-like growth factors (IGF-I and IGF-II), IGF-binding proteins (IGFBP-3, IGFBP-2), leptin and anthropometric parameters (height, body mass index) during antileukaemic treatment in children. AB - PURPOSE: The aim of the study was to estimate the anthropometric parameters and their relationship to serum levels of IGF-I, IGF-II, IGFBP-3, IGFBP-2 and leptin before and during intensive antineoplastic treatment for acute lymphoblastic leukaemia in children. MATERIAL AND METHODS: In 46 children in median age 6.6 years (range from 1.6 to 16) we evaluated at the time of diagnosis, after protocol I and after intensive treatment, height, body mass index (BMI) and IGF I, IGF-II, IGFBP-3, IGFBP-2 and leptin. RESULTS: Height SDS lowered in successive points of analysis whereas BMI SDS rose after protocol II. IGF-I SDS was low and similar at each point, IGF-II SDS and IGFBP-3 SDS values augmented progressively and IGFBP-2 SDS was significantly elevated before treatment and lowered (but not normalized) during the therapy. Leptin SDS was elevated, especially after protocol I. CONCLUSION: Leukaemia and its treatment affect directly growth factors, its binding proteins and leptin production leading to growth retardation and overweight. PMID- 16358967 TI - Soluble form of TRAIL, Fas and FasL in the serum of patients with B-CLL. AB - PURPOSE: Although many studies demonstrated expression of TNF family members in the course of B-CLL, there is a little known about relationships between soluble forms of these proteins. Furthermore, there is no study reported on effects of used therapy on this relation. The present study was designed to asses the relationships between the serum concentrations of sFas, sFasL and sTRAIL in patients with B-CLL regarding their correlation with clinical stage and used therapy. MATERIAL AND METHODS: We studied 40 patients with B-cell chronic lymphocytic leukemia (B-CLL) at diagnosis, before treatment and four weeks after therapy. To measure sFas, sFasL and sTRAIL levels in serum commercially available ELISA kits were used. RESULTS: We found increased concentrations of sFas in sera of all patients with B-CLL before treatment in comparison to the control group. There were no significant differences in concentrations of sFasL and sTRAIL between patients and control group. Increased sFasL concentrations after FC and CC therapy as well as decreased concentrations after 2CdA therapy in comparison to values before treatment were found. The concentrations of sTRAIL after FC and CC therapy were higher than those in patients before treatment. CONCLUSIONS: Results obtained suggest that relationship between sFas, sFasL and sTRAIL in sera of patients with B-CLL before treatment may facilitate the growth B leukemic cells. Changes in these relations after therapy with FC and CC can make a contribution to inhibit B cells growth on the apoptosis way in this patient group. PMID- 16358969 TI - Patient acceptance of diagnostic laparoscopy. AB - PURPOSE: To assess patient acceptance of diagnostic conventional laparoscopy and minilaparoscopy under sedoanalgesia. MATERIALS AND METHODS: 120 consecutive patients undergoing diagnostic laparoscopy were enrolled prospectively in this study. Within the first week after diagnostic laparoscopy the patients were asked to answer a total of eight questions with regard to the acceptance of the procedure. RESULTS: The inconvenience of laparoscopy was assessed with a mean of 1.6 on a scale from 0 to 10 (0 = no inconvenience, 10 = very unpleasant). The discomfort in the two days following laparoscopy were graded with a mean of 2.1 on a scale from 0 to 10 (0 = no inconvenience, 10 = very unpleasant). There was no difference between conventional laparoscopy and minilaparoscopy. Only 10% of the patients described laparoscopy more inconvenient in comparison to diagnostic gastroscopy, whereas 29% of the patients assessed diagnostic gastroscopy more inconvenient. CONCLUSIONS: Diagnostic laparoscopy under sedoanalgesia is a very well tolerated procedure. There is no difference between conventional laparoscopy and minilaparoscopy. PMID- 16358970 TI - Staphylococcus aureus septicemia in non-neutropenic adult patients hospitalized in internal medicine units. AB - PURPOSE: Staphylococcus aureus septicemia (SAS) is usually described in immunocompromised patients and during serious weakening diseases, associated with a neutropenic condition. Over the last recent years, clinic relevance of SAS has become more prominent owing to the progressive rise of methicillin-resistent strains in hospital-acquired infections and to its development in non-neutropenic patients. MATERIAL AND METHODS: The aim of our study was to evaluate the clinical features and outcome of non-neutropenic patients with positive blood culture for Staphylococcus aureus (SA) hospitalized in Internal Medicine Wards of our hospital during 1 year of observation. 24 patients with those characteristics were retrospectively recruited; five of them were then excluded from the analysis because of concomitant oncohematologic disease. The median age of the study group of patients (19 cases) was 56 years (range 18-87); 10 (52.6%) patients were male. RESULTS: Infection was hospital-acquired in 10 patients (52.6%). Predisposing factors were: central venous catheter (CVC) (47.4%), recent surgical intervention (21.0%), drug-addiction (15.8%). Main comorbidities were diabetes mellitus in 10 patients (52.6%), heart disease in 4 (21.0%), chronic renal failure in 3 (15.8%), cerebral vascular disease in 3 (15.8%). Fever >38 degrees C was found in all patients at the moment of SA isolation in blood culture. SA isolated-strains were methicillin-resistant in 7 patients (36.8%). Complications of bacteremia were: pneumonia in 4, endocarditis in 3, vertebral osteomyelytis in 2, septic splenic embolization in 1 and endophtalmitis in 1 patient. The septicemia-attributable mortality was 36.8% (7 patients). CONCLUSIONS: SAS in non-neutropenic patients observed in Internal Medicine Units are associated with significant morbidity and mortality, closer to that reported for neutropenic illnesses. PMID- 16358971 TI - The effect of green tea on the activity of aldehyde dehydrogenase (ALDH) in the liver of rats during chronic ethanol consumption. AB - PURPOSE: Alterations in the redox state during chronic ethanol consumption are associated with the oxidation of ethanol via alcohol and aldehyde dehydrogenase. Among various antioxidants present in food, strong antioxidative effects have been attributed to polyphenols of green tea. The aim of the present study was to investigate the effect of green tea consumption during chronic ethanol intake on the activity of aldehyde dehydrogenase in the liver of rats during maturation and aging. MATERIALS AND METHODS: The activity of ALDH was measured in the livers of rats aged 2 (young), 12 (adult) and 24 months (old). The rats were fed with a control liquid Lieber DeCarli diet, control liquid diet containing green tea (3 g/l), ethanol liquid diet (with increasing ethanol dose from 2.3% to 7%) and ethanol liquid diet containing green tea. RESULTS: Chronic ethanol consumption significantly increased the liver ALDH activity in young and adult rats but decreased this activity in old animals. The drinking of green tea did not alter ALDH activity in ethanol-consuming rats. Drinking green tea alone significantly increased ALDH activity in young and adult rats but did not alter this activity in old rats. CONCLUSIONS: These results demonstrate that green tea administered during chronic ethanol consumption does not prevent the changes in the hepatic ALDH activity in the rats at each age. PMID- 16358972 TI - Effects of truncal or highly selective vagotomy on the electron microscopic feature of the rabbit pancreas. AB - PURPOSE: To perform a comparative study of two types of vagotomy: trunctal vagotomy (TV) and highly selective vagotomy (HSV) effects on pancreatic morphology assessed with light microscope and ultrastructural changes assessed with electron microscope in rabbits 30, 90 and 180 days after the vagotomy. MATERIAL AND METHODS: The experiments were carried out on 89 male Popielno white rabbits, aged 4-6 months, 2.5 to 3.0 kg of body weight. On the 30th, 90th and 180th day after the vagotomy, rabbits were sacrificed and the pancreases were taken for the light and electron microscopic evaluation. RESULTS: The regressive changes coexisting with adaptive like renewal of epithelial cells and mild interstitial fibrosis resulting from vagotomy were more pronounced in the early post-operative period and tended to normalize in the later post-operative one. All the changes seen in post-operative period were more prominent after TV than after HSV. CONCLUSIONS: Both truncal and highly selective vagotomy affects evidently the morphology and ultrastructure of the pancreas in rabbits however the changes after the first procedure were more advanced than after the latter one. The intensity of the changes is highest early after these operations and tend to normalize in the later post-operative period. PMID- 16358974 TI - Human red blood cells' physiological water exchange with the plasma. AB - In the present paper, fundamental issues related to the mechanisms of human red blood cells' physiological water exchange with the plasma (for the stationary conditions) have been discussed. It has been demonstrated, on the basis of mechanistic transport equations for membrane transport that red blood cells are capable of exchanging considerable amounts of water with the plasma. Water absorption is osmosis-driven, and its removal occurs according to the hydromechanics principle, i.e. is driven by the turgor pressure of red blood cells. This newly-acquired knowledge of these issues may appear highly useful for clinical diagnosis of blood diseases and blood circulation failures. PMID- 16358973 TI - The cumulative effect of nuclear factor-kappaB (NF-kappaB) inhibition and endothelins in early cerulein-induced acute pancreatitis in rats. AB - PURPOSE: To assess effects of NF-kappaB activation inhibitor (pyrrolidine dithiocarbamate--PDTC) alone or with endothelins (ET-1, ET-2, ET-3) in early course of cerulein-induced acute pancreatitis (AP) in rats. MATERIAL AND METHODS: After 4 h of AP in Wistar rats, treated with PDTC 10 or 40 mg/kg or with PDTC 10 mg/kg and ET-1, ET-2 or ET-3, 0.5 or 1.0 nmol/kg twice i.p. in 1 h interval, free active trypsin (FAT), total potential trypsin (TPT) and lipase in 12000 x g supernatants of pancreatic homogenates, plasma alpha-amylase and histological changes were assayed. %FAT/TPT was an index of trypsinogen activation. RESULTS: %FAT/TPT significantly increased to 12.42 +/- 2.14%, lipase to 5.51 +/- 0.84 U/mg protein and alpha-amylase to 28.5 +/- 5.61 U/mL in AP vs 1.96 +/- 0.31%, 1.29 +/- 0.11 U/mg and 5.80 +/- 1.38 U/ml in healthy control. Higher dose PDTC attenuated trypsinogen activation to 3.01 +/- 0.53% and alpha-amylase to 15.3 +/- 1.38. PDTC and ET-1 attenuated %FAT/TPT to 2.55 +/- 0.18% with lower and 2.34 +/- 0.44% with higher dose. ET-3 was less effective than ET-1: 6.76 +/- 0.46% with lower dose. Lower doses of ET-1 and ET-2 with PDTC, diminished lipase activity to 2.60 +/- 0.36 and 2.94 +/- 0.33. CONCLUSIONS: Cumulative attenuation of trypsinogen activation after lower dose of PDTC and ET-1 approximated the effect of higher dose of PDTC. Additional effect of ET-3 was weaker than ET-1, and ET-2 was ineffective in this respect. The combination of this NF-kappaB activation inhibitor and ET-1 could be beneficial in early course of edematous AP by attenuating of trypsinogen activation. However, it should be treated with caution because of some unfavorable effects on histological scores of pancreatic injury. PMID- 16358975 TI - The force-frequency relationship in human heart failure: effect of pyruvate and isoproterenol. AB - PURPOSE: The purpose of present study was to investigate the effect of metabolic substrate pyruvate and beta-adrenergic agonist isoproterenol and combination of these agents on the force- and relaxation-frequency relationship in human heart failure. MATERIAL AND METHODS: The experiments were performed on isolated human ventricle strips from patients undergoing cardiac corrective open heart surgery, using conventional method of registration of electromechanical activity. The stimulation frequency of myocardial strips was 0.2, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 Hz. RESULTS: In control, i.e. at perfusion of myocardial strips by Tyrode solution and stimulation frequency 1 Hz, the contraction force (F) was 0.94 +/- 0.18 mN, half time of relaxation (tr)--178.8 +/- 9.3 ms (n= 12). Pyruvate (10 mmol/L) increased F to 176.0 +/- 13.4%, tr--104.6 +/- 3.1% (n=8, p<0.05) vs control. By the action of isoproterenol (10(-5) mol/L) F increased to 122.1 +/- 10.2%, tr decreased to 58.9 +/- 3.1% (n=4, p<0.05) vs control. The relationship of F and tr from stimulation frequency in the absence of pyruvate and isoproterenol was negative. Pyruvate and isoproterenol didn't alter the shape of force-frequency relationships but F was augmented at all stimulation frequencies. The positive inotropic effect of isoproterenol was potentiated by pyruvate. CONCLUSIONS: Pyruvate and isoproterenol alone can improve cardiac contractility in wide-range of stimulation frequency. The combination of these inotropic agents results in even more effective increase of contractile performance and therefore may be of therapeutic value in heart failure. PMID- 16358976 TI - Changes of beta2-adrenergic stimulation induced by hyperosmosis in human atrium. AB - PURPOSE: The purpose of the present study was to determine whether extracellular osmotic pressure modulates beta2-adrenergic stimulation of the contraction force and L-type Ca2+ current in human atrial myocytes. MATERIAL AND METHODS: Experiments were performed on human atrial trabeculae and myocytes isolated from the right atrium. The concentration dependent effect of salbutamol (SAL), a beta2 adrenoreceptor agonist, on peak tension (P) and L-type calcium current (ICaL) under isoosmolar (345 mOsm) and hyperosmolar (405 or 525 mOsm was achieved by adding of mannitol) conditions was studied. RESULTS: Salbutamol (10 nmol/L-10 micromol/L) added to the control solution increased P by 180.6 +/- 45.8% over control with a half-stimulation constant EC50 = 27 +/- 6 nmol/L. Under isoosmolar conditions SAL (0.1/10(3)nmol/L) increased ICaL by 182.3 +/- 19.8% over control with an EC50 2.9 +/- 0.9 nmol/L. In hyperosmolar solutions the same concentrations of SAL increased P and ICaL by 57.2 +/- 12.6% and 217.2 +/- 70.5% over control with EC50 = 640 +/- 260 nmol/L and 12 +/- 5 nmol/L respectively. CONCLUSIONS: These results indicated that hyperosmolarity reduced the effect of beta2-adrenergic stimulation, i.e. the dose-response curve of salbutamol on L type calcium current was shifted to the higher concentration range and maximal increase in contraction force was diminished in human atrial cells. PMID- 16358977 TI - Blood oxygen-carrying function during the oxidative stress induced by lipopolysaccharide with a modification of the L-arginine-NO pathway. AB - PURPOSE: Our aim was to study the blood oxygen-carrying function during the oxidative stress with a modification of the L-arginine-NO pathway. MATERIAL AND METHODS: Oxidative stress was induced by intravenous administration of Escherichia coli lipopolysaccharide (LPS) to rabbits. To modify the L-arginine-NO pathway, animals were administered with NG-nitro-L-arginine methyl ester intravenously 60 min after the LPS. Mixed venous blood was sampled for evaluation of blood oxygen transport before and at 120 and 240 min after the LPS administration; tissue samples (heart, lung, liver, kidney and muscle) were also prepared. The following parameters were measured hemoglobin-oxygen affinity, concentrations of conjugated dienes, Schiff bases, alpha-tocopherol and activity of catalase. RESULTS: During the NO synthase inhibition the oxidative stress was characterized by a shift of hemoglobin oxygen dissociation curve rightwards, more prominent activation of lipid peroxidation and decreased tissue levels of antioxidant defense factors. CONCLUSIONS: The inhibition of NO generation induces a shift of prooxidant-antioxidant balance--obviously, not only due its potentially high levels and reactivity with the various target molecules (with a development of oxidative stress), but also because of the lower contribution of other factors including the hemoglobin-oxygen affinity change into the body antioxidant potential. PMID- 16358978 TI - Diurnal rhythm of blood pressure, heart rate and adrenergic activity in patients with normotension treated with continuous ambulatory peritoneal dialysis and haemodialysis. AB - PURPOSE: People with normotension and with essential hypertension are subjected to the diurnal rhythm of blood pressure (BP) with higher values during the day than during the night. Among dialysed patients nocturnal reduction of BP is blunted. The aim of the study was to evaluate diurnal BP rhythm and adrenergic activity measured as values of catecholamines. MATERIAL AND METHODS: Study was performed among dialysed patients with normotension: 13 haemodialysed patients (HD), 8 patients treated by continuous ambulatory peritoneal dialysis (CAPD) and 10 controls (C). Ambulatory BP monitoring (ABPM) was done by using Micro SJ7400 AMP device. Catecholamines concentrations were measured by HPLC-ED method before and after cold pressure test. RESULTS: There was no significant difference between manual measurements of BP done by dialysis nurses and mean values of 24 hours ABPM in CAPD group and C group and 48-hours ABPM among HD patients. Diurnal BP was blunted in 80% of HD patients during the day of haemodialysis, 70% during the day without haemodialysis and in CAPD group in 50%. Heart rate (HR) variability was comparable in HD and CAPD groups and significant lower than in C group. Baseline noradrenaline (NA) as well as NA (ng/ml) post cold pressure test levels were significantly higher among HD patients (463 +/- 21, 546 +/- 31) and CAPD patients (452 +/- 76, 527 +/- 92) as compared with C (206 +/- 53*, 315 +/- 61*). (x +/- SD), *p<0.001 CONCLUSIONS: Despite increased adrenergic activity and altered diurnal rhythmof BP and HR exist in dialysed patients we didn't find directly relationship. Another or composed factors couldaffect diurnal rhythm of BP and HR. PMID- 16358979 TI - Cholecystokinin octapeptide (CCK-8) concentration in plasma is not affected in functional abdominal pain in children. AB - PURPOSE: Cholecystokinin regulates gut motility and visceral sensation. The aim of the study was to determine the diagnostic value of plasma cholecystokinin octapeptide (CCK-8) concentration in children with functional abdominal pain (FAP). MATERIAL AND METHODS: Fifty-two children (33 girls and 19 boys) aged 6-17 years with chronic abdominal pain were included in this study. On the basis of clinical data, results of endoscopy and Criteria for Functional Disorders the patients were divided into three groups: group 1--functional dyspepsia (FD), group 2--irritable bowel syndrome (IBS), group 3--non-specific FAP. The control group consisted of children without abdominal pain in anamnesis. CCK-8 concentrations in plasma were measured with radio immunoassay technique, after plasma extraction. In study protocol we analysed CCK-8 levels in fasting state and 15, 30, 60 minutes after a standard test meal. RESULTS: In the fasting state plasma levels of CCK-8 were similar in each group and in controls. In the IBS patients CCK-8 levels were not increased after meal. In groups 1, 3 and controls postprandial levels were higher when compared to fasting state (p<0.05). Area under curve of CCK-8 plasma concentration was the lowest in group 2, but not significant compared to controls and other groups. No correlation was found between main symptoms of FD and IBS and CCK-8 concentration in plasma. CONCLUSIONS: We conclude that gut dysmotility and symptoms of functional abdominal pain in children are not concerned with alteration of plasma CCK-8 levels before and after meal. PMID- 16358980 TI - Atopy patch test in the diagnosis of food allergy in children with atopic eczema dermatitis syndrome. AB - PURPOSE: Food allergy has been demonstrated to play an important role in the pathogenesis of atopic eczema dermatitis syndrome (AEDS), affecting often atopic infants and young children. The most commonly offending foods are cow's milk, hen's egg, wheat and soy; implicating immediate (IgE-mediated) and late-phase (T cells) immunological reactions in the pathogenesis of skin lesions. The diagnostic work-up of suspected immediate food reactions includes skin prick tests (SPT) and the measurement of food-specific antibodies (sIgE). The methodology of atopy patch test (APT) has been reported as a diagnostic tool with high predictive capacity for late-phase clinical reactions in children with atopic dermatitis. Although APT has been introduced into the diagnostic procedure for food allergy, its diagnostic accuracy remains still controversial; especially in older children. The aim of study was to evaluate the diagnostic accuracy of the atopy patch test in the detection of food allergy in correlation with SPT, sIgE and positive oral food challenge to milk, in children suffering from AEDS and to assess the sensitivity and specificity of this method in dependence on the age of investigated children. MATERIAL AND METHODS: 34 children (25 boys, 9 girls) aged 5 months-16 years with suspicion of milk-related AEDS were investigated. These patients were subdivided into 2 age groups: group A--20 children (<3 years), group B--14 children (>3 years). The diagnostic procedures as skin-prick tests and atopy patch test were performed. The specific IgE to cow's milk allergens were also measured. The open and blind diagnostic oral food challenge were performed to verify the results of tests. Sensitivity, specificity, positive (PPV) and negative (NPV) predictive value of APT were calculated in both age groups. RESULTS: A positive challenge response to milk was found in 65.0% of investigated children in group A and in 35.7% in group B. No statistical differences in the prevalence of immediate (p<0.1905) and delayed type (p<0.409) reactions has been found between age groups. Positive APT to milk were noticed in 55.0% of patients in group A and in 35.7% of children from group B, that has been in correlation with positive delayed-type reactions in oral food challenge in 72.7% and 80.0% in corresponding age groups. Polysensitization to other food allergens confirmed by SPT and/or sIgE was detected in 35.0% of patients younger than 3 years of age and in 50.0% of older children. The prevalence of positive APT to other foods (soy, rice, maize, cereals) was significantly higher (p<0.0073) in the polysensitized children from group A. Sensitivity of SPT/sIgE in children with immediate-type reactions to milk was 100%, specificity 94%. Sensitivity of APT to cow's milk in children with late phase reactions was 80% in both age groups; specificity 70%/89% with comparable PPV in both groups (73%/80%). Parallel skin testing with combined patch test and evaluation of sIgE enhanced the value of sensitivity to 92% in the group A and specificity to 89% in the group B. For PPV corresponding figures were 85%/80%. CONCLUSIONS: APT was found to be more sensitive and specific method than SPT/sIgE in diagnosing delayed food allergy in children with AEDS. No age correlation between positive results of APT and oral food challenge and higher specificity of APT in older children confirm its accuracy in diagnosing delayed cow's milk allergy in all age groups of children. Combined skin prick and patch testing significantly enhances identification of food allergy in children with AEDS. The outcome of the APT with food does not seem to be influenced by age of children, but because of its variability of sensitivity and specificity, a diagnosis of food allergy should be confirmed by oral food challenge. PMID- 16358981 TI - Immunoblotting in the diagnosis of cross-reactivity in children allergic to birch. AB - PURPOSE: The scientific experiments with new immunological methods (immunoblotting, RAST inhibition) and isolation of recombinant allergens suggest structural similarities in the allergenic components responsible for cross reactions. Immunochemical and molecular biology studies indicate that epitopes of major allergen (Bet v 1, Mal d 1) contain more IgE binding epitopes than minor allergens (Bet v 2, Mal d 2), what explained clinical importance of major birch and apple allergens, but it is individual. The important role in cross-reactivity play also proteins with low molecular weight; a potentially dangerous allergen is lipid transfer protein (LTP) inducing severe systemic reactions in allergic subjects. The recent studies indicate that the IgE cross-reactivity patterns and the clinical relevance is still not clear and that only some of patients with confirmed IgE cross allergy to Bet v 1 and Mal d 1 demonstrated clinical symptoms after ingesting of apple. The aim of study was to establish the pattern of cross reactivity between major (Bet v 1) and minor (Bet v 2) birch pollen allergens and apple proteins in children allergic to birch using recombinant allergens and immunoblotting method. MATERIAL AND METHODS: The prospective study were carried out on the group of 13 children aged 4-16 years, referred to the IIIrd Department of Paediatrics in Bialystok and outpatient clinic with clinical symptoms of food and inhalant allergy. Inclusion criteria to the study were: allergy to birch pollen recombinant allergens and apple, confirmed by presence of specific IgE in the sera of patients. The allergens from peel and pulp of apple and birch were separated and loaded onto the polyacrylamide electrophoretic gel and than transferred to membranes by Western blotting. Antigen-IgE complex was detected using goat anti-human IgE antibodies labelled with alkaline phosphatase. RESULTS: Only few sera presented strong reactions in immunoblotting to birch pollen proteins with a molecular weight of 17-18 kDa, corresponding to the main birch allergen Bet v 1. Most of sera having positive reaction vs Bet v 1 showed cross reactivity with Mal d 1. All sera recognized specifically the main allergen of apple peel Mal d 3 with molecular weight < 10 kDa (Lipid Transfer Protein). CONCLUSIONS: Immunoblotting method allows to verification of cross-reactivity recognized by presence of specific IgE. The nature of proteins responsible for sensitization can influence the spectrum of offending foods and the clinical features of allergic reactions. PMID- 16358982 TI - Hypersensitivity to hydrolyzed cow's milk protein formula in infants and young children with atopic eczema/dermatitis syndrome with cow's milk protein allergy. AB - PURPOSE: Atopic eczema/dermatitis syndrome (AEDS) is often the first manifestation of atopic disease in children. Food hypersensitivity should be considered in approximately 40% of these patients. AEDS children with cow's milk allergy are commonly prescribed a hydrolyzed formulas or amino acid-based formulas for an alternative protein source. The aim of this study was to investigate hypersensitivity to extensive hydrolyzed casein and whey proteins in AEDS children with cow's milk protein allergy (CMA). MATERIAL AND METHODS: The study included 67 hospitalized children with AEDS (m/f--43/24), aged 1-28 months (mean 11.34 +/- 8.52) and CMA confirmed by oral food challenge. All patients were treated with extensively hydrolyzed formulas: 48/67 children with casein hydrolysates and 19/67 children with whey hydrolysates. RESULTS: In most of studied children we recognized severe AEDS (SCORAD Index: mean 55.41 +/- 17.4; 95% CI 51.17-59.66) with elevated total IgE (mean 432.98 +/- 1030.46; 95% CI 181.63-684.33). In 22/67 children (32.8%) we established diagnosis of hypersensitivities to hydrolyzed formula (HHF): in 17/22 to casein hydrolysates, in 4/22 to whey hydrolysates and in 1/22 to amino-acid based formula. Children with HHF did not differ in the severity of AEDS evaluated by SCORAD (57.18 +/- 16.59 vs 54.56 +/- 17.90), the serum level of total IgE (603.9 +/- 1253 vs 349.4 +/- 906.1) and the time of breast-feeding (4.4 +/- 4.0 months vs 6.8 +/- 7.28). They differ in the number of plasma eosinophils and positive correlation between number of eosinophils and serum level of total IgE (p<0.05, r=0.46 vs r=0.07). CONCLUSIONS: Children with moderate or severe atopic eczema/dermatitis syndrome can demonstrate hypersensitivity to hydrolyzed formula recommended for therapeutic indications. PMID- 16358983 TI - Acute phase proteins serum concentrations in children are related to urinary iodine excretion. AB - PURPOSE: The paper presents links between iodine provision and selected acute phase proteins' (APP) serum concentrations as well as their glycosylations profiles (investigated with the use of affinity immunoelectrophoresis with Concanavalin A as ligand) in children. MATERIAL AND METHOD: 116 children (58 girls and 58 boys) were enrolled. Iodine level was measured in the morning (7:30 8:30) urine portion, using Cr-As method. According to iodine level children were divided into two groups. The first one consisted of 56 children with decreased iodine level (lower than 100 micrograms/L), second--60 children with iodine level higher than 100 micrograms/L. In serum the concentration of ferritin, beta2 microglobulin (beta2-MG), thyroxin (T4), triiodothyronin (T3), thyrotrophic hormone (TSH) were measured by radioimmunoassay (BELORIS, Belarus). Concentrations of APP: C-reactive protein (CRP), alphal-acid glycoprotein (AGP), alphal-antichymotrypsin (ACT), alphal-antitrypsin (AT), haptoglobin (Hp), alpha2 macroglobulin (A2-M), ceruloplasmin (Cp) and transferrin (Tf) were measured in sera samples by rocket immunoelectrophoresis acc. to Laurell with antibodies and standard from DakoCytomation, Denmark. Microheterogeneity of AGP, ACT and Tf was estimated using affinity immunoelectrophoresis with ConA as a ligand, acc. to Bog Hansen. RESULTS: It was established, that CRP level was lower than upper limit of normal range. Levels of other investigated proteins were reliably dependent on the level of iodine. Especially for AGP lower level was observed for children of the group with low iodine level. In children with low iodine level along with the decrease of serum AGP concentration altered glycosylations profile was observed, namely decrease in the content of variant non-reactive to ConA (W0) and increase in content of weakly reactive (W1) and reactive (W2) variants content, which resulted in increase of the reactivity coefficient (AGP-RC). Similar tendency in alterations of distinctly glycosylated variants in relation to iodine level could be shown for ACT. Serum concentration of any investigated protein was not dependent on the concentration of the hormones of pituitary-thyroid system. CONCLUSIONS: It seems that the influence of the iodine level is direct, not via thyroid hormones. It could be suggested that in euthyroid children with low iodine excretion with urine a hidden iodine deficiency is already registered by the regulatory mechanisms and a kind of acute phase reaction is started, may be in order to increase iodine uptake and storage. PMID- 16358984 TI - Extraction of nerve cells in images with herpetic infections. AB - In this paper, algorithms for extracting and diagnosis of nerve cells with herpetic infections are proposed. Degree of herpetic lesions is divided into four classes. Morphological characteristic of herpetic lesions for cell is accomplished by analyze cells structure. Because virus of herpes change shape of nucleus. Therefore automated analysis of herpetic lesion carry out by morphology segmentation and cells structure identification. The algorithms have been successfully used in practical systems and showed good results. PMID- 16358985 TI - Alterations of lymphocyte subpopulations in choroidal melanoma patients undergoing surgery. AB - PURPOSE: The alterations of lymphocyte subpopulations assessment after surgery in choroidal melanoma patients compared to cataract patients. MATERIAL AND METHODS: 12 patients with malignant melanoma of the choroid, 10 patients subjected to surgery due to cataract. Methods--flow cytometric measurement of absolute lymphocyte count, the number of all T cells (CD3+), T helper lymphocytes (CD3+ CD4+), T cytotoxic lymphocytes (CD3+CD8+), B lymphocytes (CD19+), NK cells (CD3 CD16+) and T cells (CD3+) cells with gammasigma TCR, on the day of surgery and two days after it. RESULTS: Comparable numbers of cells were observed in both groups prior to surgery, but the behavior of some populations differed: CD3+, CD3+CD4+ cells increased in melanoma patients whereas they decreased in reference group, the number of T lymphocytes with gammasigma TCR was significantly higher in melanoma patients before surgery and it did not differ after it. CONCLUSIONS: Though there were no significant differences in lymphocyte subpopulations between melanoma patients and the reference group, it seems that the presence of tumour influences the reactivity of the immune system to the trauma (surgery). PMID- 16358986 TI - Phagocytic and bactericidal activity and morphological parameters of blood platelets in patients with Trichinella spiralis infection. AB - PURPOSE: The production of IgE increases in parasitic invasions, triggering local or systemic inflammatory response with the involvement of blood platelets. The aim of the study was to assess the number and morphological parameters of blood platelets as well as their phagocytic and bactericidal activity in patients with Trichinella spiralis infection. It is interesting to investigate the blood platelet response following Trichinella spiralis in order to elucidate possible effects on non-specific immunity. MATERIAL AND METHODS: Twenty-six patients with Trichinella spiralis (before and after antiparasitic therapy) and forty healthy subjects were examined. The platelet count and morphological parameters were determined using a hematologic analyzer Technicon H-1 System. The platelet phagocytic activity was determined by measuring the percentage of phagocytizing cells and the phagocytic index. The bactericidal activity was assessed measuring the percentage of the bacteria killed by platelets and plasma. The strain Staphylococcus aureus ATCC 6538P was used for this purpose. RESULTS: In patients infected with T. spiralis morphological parameters do not change, except for the percentage of large platelets. In the course of trichinellosis the phagocytic index of platelets is statistically significantly decreased and platelet bactericidal activity is impaired, while the bactericidal activity of the plasma is statistically significantly increased, compared to healthy subjects. CONCLUSIONS: The present study has revealed that due to T. spiralis infection, the percentage of large, young blood platelets is decreased. The parasitic infection causes impairment of non-specific immunity through decreased bactericidal activity of blood platelets. PMID- 16358987 TI - Development and psychokinetic therapy of children suffering from West Syndrome- an overview. AB - Paper presents reasons leading to the West Syndrome, disturbances occurring in the child's development and possible rehabilitation programmes. Psychological rehabilitation of a child with West Syndrome is multilateral, but its main aim is to improve the quality of life, social adaptation and optimizing of the cognitive functioning. Taking into consideration the psychokinetic retardation occurring in the West Syndrome because of prenatal, perinatal or postnatal disturbances apart from pharmacotherapy an intensive psychokinetic stimulation based on plasticity of the brain should be stressed out. Only the complex, intensive and long lasting rehabilitation of a child may secure to it the best possible development. PMID- 16358988 TI - The duration of breastfeeding and attention deficit hyperactivity disorder. AB - PURPOSE: To examine whether duration of the breastfeeding is associated with the symptoms of attention deficit hyperactivity disorder in children. MATERIAL AND METHODS: A total of 100 children aged 4-11 years were divided into two groups: 60 children with ADHD symptoms (based on ICD-10) and 40 subjects of normal control grup. The structured interview and the retrospective questionnaire (including the items: number of pregnancy, pregnancy course, gestational age, status of newborn, birth weight, duration of breastfeeding: <3 months; 3-6 months; 6-12 months; >12 months) were used during the study of the both examined groups to indicate the risk factors of development. RESULTS: No significant differences in the percentages of duration of pregnancy, pregnancy complications, delivery complications, condition of the newborn, and birth weight were found between the two groups. The mean of the duration of breastfeeding for group with ADHD was 0.45 year: 5 months and 9 days (median 0.25 year: 3 months). The mean of the duration of breastfeeding of control group was 0.55 year: 6 months and 18 days (median 0.46 year: 5 months) and was significantly greater than that of group with ADHD (p<0.04). The 36 (60%) children with ADHD were breast fed less than 3 months. For comparison 13 (32.5%) controls were breast fed less than 3 months. Significant differences were found among the two children groups (p < 0.05). CONCLUSIONS: The short duration of breastfeeding as environmental factor may be considered a risk factor of ADHD symptoms. However, the further studies are nedeed. PMID- 16358989 TI - Chlamydia trachomatis infection in chronically hemodialyzed patients. AB - PURPOSE: In the general population there is association between Chlamydia trachomatis (Ch. trachomatis) infection and reactive arthritis (RA). RA is a systemic illness characterized by inflammatory synovitis. Arthritis tends to be oligoarticular and involves mainly the lower limbs. The aim of this study is to assess the age and sex specific prevalence of Ch. trachomatis infection in dialysis population and to find possible relationship between manifestation of infection and renal osteodystrophy. MATERIAL AND METHODS: The study was conducted in 53 patients: 22 women (W) and 31 men (M), with a mean age of 58.1 +/- 15 years, treated with HD for 28.5 +/- 28.2 months. The Ch. trachomatis infection was assessed by the detection IgG antibodies for Ch. trachomatis. Also some other biochemical parameters of osteodystrophy, inflammation and malnutrition were measured. RESULTS: The presence of a high titre of anti-Ch. trachomatis antibodies was found in 22 patients--41% [G IgG (+)]. Mean level of anti-Ch. trachomatis antibodies was significantly higher in G IgG (+) than in seronegative patients [G IgG (-)]: 19.0 +/- 8.6 vs 4.0 +/- 2.1 U/ml, p<0.001. There was no difference in mean age of seropositive and seronegative patients for Ch. trachomatis (62.4 +/- 13.1 vs 56.2 +/- 15.9 years). We did not observe in both groups of patients any differences in mean level of C-reactive protein (CRP): 12106.2 +/- 10791.0 vs 14015.3 +/- 11194.3 ng/ml. The mean ferritin level was significantly higher in G IgG (+): 624.3 +/- 375.7 vs 418.3 +/- 341.4 ng/ml, p<0.05. Significant negative correlations were found in G IgG (+) between IgG antibodies and transferrin saturation (r=-0.645719, p<0.001) and between CRP and calcium (r=-0.4526, p<0.05). IgG antibodies were detected frequently in W (60%) than in M (29%). Mean level of IgG was significantly higher in seropositive W than in seropositive M (23.3 +/- 7.8 vs 12.1 +/- 4.2 U/ml, p<0.0001). The seropositive W were older (67.9 +/- 11.8 vs. 53.8 +/- 11.0 years, p<0.02) and seropositive W were shorter treated with HD (18.1 +/- 16.6 vs 43.7 +/- 30.6 months, p<0.02). The mean serum calcium conc. and phosphorus were significantly lower in seropositive W (2.1 +/- 0.1 vs 2.3 +/- 0.2 mmol/l, p<0.05 and respectively 1.3 +/- 0.3 vs 1.8 +/- 0.2 mmol/l, p<0.005). Likewise the mean transferrin saturation (TS) was significantly lower in that group (25.7 +/- 7.3 vs 38.0 +/- 11.3%, p<0.01). There were no differences between seropositive men and women in mean serum concentrations of CRP, iPTH, albumin and hemoglobin. We found in seropositive W significant negative correlation between IgG antibodies and age (r=-0.633, p<0.02). CONCLUSIONS: The patients treated with HD were quite frequently shown significantly elevated level of IgG antibodies for Ch. trachomatis. It could have be connected with past infection. The antibodies were more commonly detected in women, particularly in younger patients. No relationship between osteodystrophy and Ch. trachomatis infection was found. PMID- 16358990 TI - Increased serum levels of troponin I and lesions in coronary angiography in hemodialysed patients. AB - PURPOSE: Calcium-phosphate disorders are a frequent finding in HD patients, and, in some cases, may cause an abnormal coronary calcification. Some of the HD patients have increased serum cTnI level without evidence of acute coronary syndrome. The aim of this study was to determine if there is a correlance between increased cTnI levels and presence of stenotic changes in coronary arteries in asymptomatic HD patients. MATERIAL AND METHODS: In 13 of 119 HD patients (M:F 10:3) a coronary angiography was performed. The mean age of the patients was 53 years (33-76) and the mean HD duration was 55 months (3-156). cTnI was analyzed by AxSYM system and, subsequently, by VIDAS system. RESULTS: A constant or intermittent elevation of cTnI was detected in 5 of 13 patients. In 10 of 13 pts a critical stenosis of at least 1 coronary artery was found. A critical stenosis was found in 4 of 5 cTnI (+) patients and in 6 of 8 cTnI (-) patients. An excess calcification of coronaries was observed in 7 patients, including 1 cTnI positive patient with no evidence of coronary stenosis. CONCLUSIONS: 1. The elevation of cTnI in asymptomatic HD patients is observed when there is: (I) excess calcification accompanied by a critical stenosis of at least 1 coronary artery, (II) a critical stenosis of 2 or more coronaries with no evidence of calcification. 2. We suggest that excess cardiovascular calcification in HD patients may be one of the major factors responsible for the troponin release. PMID- 16358991 TI - Results of improvement in adequacy of intermittent hemodialysis in uremic patients. AB - PURPOSE: Increasing number of patients, who need intermittent hemodialysis (IHD), is a great challenge for every society. The aim of study is to look if small increase in IHD adequacy is able to improve standard medical parameters. MATERIAL AND METHODS: In 40 patients, Kt/V was monitored on-line during the middle IHD session in the week, 4 times in each of 6 consecutive months. In the first month of observation Kt/V was lower (1.09 +/- 0.02) than in the further months, in which Kt/V was increasing to 1.17 +/- 0.01. Blood count was estimated every month. At the beginning of study period, after 3 months and at the end of studies, dry body mass, body mass index (BMI), the blood pH and serum concentration of calcium, phosphate, intact parathormone (iPTH), total protein, albumin, cholesterol, iron, ferritin, urea and creatinine were determined. RESULTS: The increase in Kt/V was accompanied by rising values of hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume, iron, blood pH before and after IHD session as well as by decreasing values of PTH. Statistically unchanged parameters included dry body mass, BMI, serum concentration of total protein, phosphate, cholesterol and ferritin as well as white blood cells and platelet count. There were correlations between Kt/V and serum concentrations of phosphate, PTH, ferritin, Hb and Hct, indicating that higher IHD doses were provided to patients in more advanced uremic state. CONCLUSIONS: Even small increase in IHD adequacy leads to beneficial changes in management of uremic patients. PMID- 16358992 TI - Patients' quality assessment of ambulatory obstetric and gynaecological services. AB - PURPOSE: The quality could be assessed from two perspectives: internal and external. From the internal perspective the quality means being consistent with particular conditions and standards. The external perspective is based on relative assessment of the product made by a client who is also aware of other competitors' offer. Despite the professional assessment which is focused on providing health services according to medical and managerial correctness, patient's assessment is also relevant. Measuring patient's satisfaction is additional method of health services quality assessment. The aim of the study was to estimate patients' opinion on quality of ambulatory obstetric and gynaecological services. MATERIAL AND METHODS: The study was conducted in 11 obstetric and gynaecological out-patient clinics of Lublin in September and October 2003. The study tool was an author's questionnaire. Patients were asked to assess such areas as registration before visit, their relationship with nurses and gynaecologists and other aspects of services provision like intimacy assurance and respecting Patient's Rights. The collected data was statistically analysed. RESULTS: 635 patients took part in the study. The biggest groups in the studied population were women at the age of 20-30 years, married, living in cities and secondary educated. It was found that around half of the population is satisfied and 2.2% of them are unsatisfied with the fact that they have chosen particular out-patient clinic. More than 70% of women had positive opinion of following aspects influencing general opinion about service quality: politness of reception desk staff, opening hours, the length of time before a patient is seen by the specialist, intimacy assurance and respect for Patient's Rights in the practice. 80% of patients were satisfied with the relationship with nurses and 3.3% were unsatisfied with it. Eight patients out of ten were satisfied with gynaecological care, less than 2% were unsatisfied. The studied women had also high opinion of the course of visit. More than 90% of studied patients trust their gynaecologists. CONCLUSIONS: More satisfied with the services provided by the obstetric and gynaecological out-patient clinics were women living in cities than in villages and those visiting nonpublic than public practices. PMID- 16358993 TI - Serum isoprostanes levels in patients after abdominal hysterectomy. AB - PURPOSE: Reactive oxygen species (ROS), continuously generated in tissues, are involved in signal transduction under physiological conditions. The amount of ROS increases in response to surgical trauma. Isoprostanes are novel sensitive and specific markers of lipid peroxidation in vivo. Plasma concentration of isoprostanes increases in patients with various diseases associated with oxidative stress. In the present study we investigated the effect of abdominal hysterectomy on serum isoprostanes concentration. MATERIAL AND METHODS: The study was performed in 20 women (aged 45-63, average 50.3) who had undergone total abdominal hysterectomy with salpingooophorectomy, operated for benign diseases in the 1st Department of Gynaecology of Lublin Medical University. Isoprostanes were assayed by enzyme immunoassay (EIA) using 8-isoprostane EIA kit (Cayman Chemical, Ann Arbor, MI, USA). RESULTS: Serum concentration of isoprostanes before the surgery had value 38.9 +/- 10.7 pg/ml and it decreased at 8, 24 and 96 h after the operation. CONCLUSIONS: Measurement of serum isoprostanes in small group of patients after hysterectomy did not brought the clear answer if the assessment of isoprostanes levels is a valuable method for evaluation of oxidative stress after a surgery. PMID- 16358994 TI - Binding of alpha1-antichymotrypsin to the surface of lymphocytes--preliminary study. AB - PURPOSE: The paper presents preliminary results of investigation of binding of alpha1-antichymotrypsin to the surface of peripheral blood lymphocytes. MATERIAL AND METHODS: Pooled serum samples from healthy individuals served as source of alpha1-antichymotrypsin for isolation using chromatography. Binding of alpha1 antichymotrypsin to the surface of peripheral blood lymphocytes was measured by flow cytometry. RESULTS: Even on native cells alpha1-antichymotrypsin may be detected. After incubation with isolated preparation the percentage of positive lymphocytes increased. CONCLUSIONS: The presence of alpha1-antichymotrypsin on the surface of lymphocytes may imply its regulatory role during acute phase response and early immune response. PMID- 16358995 TI - The analysis of mortality from cardiovascular diseases in Pomeranian province. AB - PURPOSE: Descriptive epidemiology characterises frequency of appearance of given event (here decease) in dependence of many factors concerning a person, region or time of existence of given salubrious phenomena. The source of information was the official death registry that provides complete records of all deaths that took place in Pomeranian province. This description of sanitary situation of people from particular area enables doing comparison between regions, facilitates researching etiological factors, planning work of medical workers and programming preventive rules. Cardiovascular diseases during last fifty years are the main reason of death of people from developed countries which is also Poland. MATERIAL AND METHODS: The aim of this work is to find out differences in health condition between citizens of Pomeranian province and other people in Poland and countries of European Union. RESULTS: In Pomeranian province in 2002 the highest mortality from cardiovascular diseases was observed for Sztum county (587.5/100,000) and was 72.5% of all deaths in this region. Similarly, there was high mortality these reasons in Tczew county (442.1/100,000), Malbork county (406.9/ 100,000) and also in Tricity (424.8/100,000). The lowest mortality from cardiovascular diseases was observed in Gdansk county (257.2/100,000) and was only 40% of all deaths in this region. Relatively low mortality was in Czluchow county (288.9/100,000). Frequency of death from cardiovascular diseases in Pomeranian province has become lower from year 2000 (361.0/100,000) to year 2002 (347.9/100,000). It was lower than in other parts of Poland (449.8/100,000 in year 2000) but higher than in countries of European Union (257.8/100,000 in year 2000). CONCLUSIONS: Mortality from cardiovascular diseases has decreased during last few years. Also there are distinctions in this phenomena among regions of Pomeranian province, other parts of Poland and countries of European Union. From these reasons health care should be differentiated to address the differences in spatial patterns of risk observed. PMID- 16358996 TI - Evaluation of secretory mucin concentration of patients with squamous cell carcinoma oral cavity. AB - PURPOSE: Secretory salivary mucins constitute a heterogenous group of glycoproteins, synthesized and secreted by submandibular, sublingual gland and small glands of oral mucosa. The most significant functions of mucins in case of oral cavity carcinoma are: participation in oral pellicle formation, lubrication and creation of heterotypic complexing. The aim of this study was to assess mucins concentration, and finally to establish the correlation between concentration of mucins in saliva and clinical advancement according to TNM. MATERIAL AND METHODS: The research was conducted on mixed resting and stimulated saliva of patients with oral squamous cell carcinoma. Mucin's concentration was measured one day before, and thirty days after surgical procedure. The volume of saliva was volumetrically determined, quantitative evaluation of mucins was accomplished by PAS method. RESULTS: In comparison with K group, a significant decrease of mucins was found in resting and stimulated saliva of patients with carcinoma in all degrees of clinical advancement. Mean value of mucin in resting and stimulated saliva after surgical treatment were lowered. The degree of carcinoma clinical advancement correlated negatively with mucin concentration. CONCLUSIONS: The decrease of mucin contained in saliva may be important in further evolution or progression of carcinoma. The results also suggest that saliva may be a significant diagnostic material in carcinoma research. PMID- 16358997 TI - Low dose rofecoxib, inflammation and prostacyclin synthesis in acute coronary syndromes. AB - PURPOSE: To assess the influence of low dose rofecoxib on inflammatory mediators and prostacyclin synthesis in patients with acute coronary syndromes (ACS) in a short-term follow up. MATERIAL AND METHODS: Twenty nine patients with ACS without ST elevation were randomized to simvastatin alone or together with low dose rofecoxib. Serum levels of interleukin 6 (IL-6), 6-keto-PGF-1alpha--stable product of prostacyclin (PGT2) and hs-C-reactive protein (hs-CRP) were assessed on enrollment and after 30-day follow up. RESULTS: Combination of rofecoxib with statin significantly decreased levels of hs-CRP after one month therapy (5.21 mg/l +/- 4.12 vs 2.11 mg/l +/- 2.1; p=0.0092). This effect was not evident in a group on statin alone (3.95 mg/l +/- 3.33 vs 2.48 mg/l +/- 2.39; p=0.31). 6-keto PGF-1alpha increased not significantly in both groups. IL-6 concentration has not changed during follow up. CONCLUSIONS: Low dose of selective COX-2 inhibitor exerts significant anti-inflammatory effect and does not diminish PG12 synthesis in study group of patients with ACS. PMID- 16358998 TI - The dentist's role in an emerging cancer treatment technology. AB - With the ADA's increasing emphasis on oral cancer detection and treatment, the following article illustrates a way in which any dentist can become involved in proton radiotherapy, a new cancer treatment technology. PMID- 16358999 TI - Socket preservation followed by dental implant supported restorative treatment: a case report. AB - Implant-supported restorative treatment continues to revolutionize all aspects of dentistry. Any clinical procedure(s) with the potential to enhance and improve the success of implant-supported restorative treatment will gain favor with both practitioner and patient. Socket preservation following tooth extraction appears to offer several clinical advantages in preparing the ridge for future implant placement. A case report is presented describing treatment of a patient with evidence of root resorption of the maxillary central incisors, resulting in extraction and socket preservation followed by implant supported restorative treatment. PMID- 16359000 TI - The status of kindergarten and middle school entry dental examinations in Indiana. PMID- 16359001 TI - Spore testing to validate heat sterilization. PMID- 16359002 TI - [Endoscopic endonasal sinus surgery using a navigation system for pituitary adenoma]. AB - Hardy's operation using a microscope has long been the standard for treating pituitary adenoma. A new endonasal approach to the sella using an endoscope combined with a navigation system has been reported, which we used to conduct endocopic endonasal hypophysectomy from October 2000 to June 2003 in 11 patients with pituitary lesions. We introduced an angle-dependent navigation system, Neuro Navigator III. We approached the sphenoid sinus mainly via the hemilateral common meatus. The deviation of the nasal septum and sphenoidal septum was carefully evaluated to determine the optimal operating side. We concluded that the hemilateral common meatus route is useful because it is least invasive in endocopic endonasal hypophysectomy. Another route should be taken, however, if hemorrhaging is uncontrollable or the tumor is quite large. Navigation systems are quite effective in executing this operation safely. Angle-dependent navigation system is a good choice for this operation, considering its cost performance. PMID- 16359003 TI - [Investigation for cervical lymph node metastasis in unknown primary sites]. AB - In patients with cervical adenopathy, especially, those of cervical lymph node metastasis with no detectable primary tumor, diagnosis and treatment planning can become confused. We evaluated 36 patients with cervical lymph node metastasis of unknown origin between 1985 and 2002. Primary sites were detected in 20 before treatment. The other 36 patients clearly had no primary lesions when treatment started. Primary sites were 5 cases of oropharynx, 2 of the parotid gland, and 1 each of larynx, nasopharynx, hypopharynx, and malignant lymphoma detected in 11 after treatment for cervical lymph nodes. No primary lesion was found in 28 patients. The neck LN stage was N1 in 11 patients, N2 in 29, N3 in 11, and unknown in 8. To detect the primary site, we conducted "random" biopsy, panendoscopy, and radiographic evaluation including FDG-PET. Biopsy sites were the nasopharynx, palatine and lingual tonsil, and piriform sinus. Some 35 patients (59.3%) underwent random biopsy, and primary sites were found this way in 5 patients (14.3%). The 36 who had no primary lesion were treated for cervical lymph nodes, of whom 24 underwent neck dissection. Chemotherapy and radiotherapy were the treatment of choice in many cases. We analyzed 31 patients for 5 year survival. Overall survival was 63.7%, disease-specific survival 69.2%, and disease-free survival 46.8%. In another analysis a statistically significant difference was seen in survival among patients who had neck surgery or not (85.7% vs. 38.9%, p = 0.029; log rank test). Analysis suggested that primary sites should be studied by CT, MRI, FDG-PET, and panendoscopy, including random biopsy. The primary site cannot be detected, treatment should initially involve cervical adenopathy with combined surgery, chemotherapy, and radiotherapy. After treatment, the patient should be followed up carefully to find the primary lesion. PMID- 16359004 TI - [Effect of the external nasal dilator breathe right and nasal cycle influence]. AB - The external nasal dilator Breathe Right is a nonprescription disposable mechanical device worn over the bridge of the nose that supports the outer walls of the nasal vestibule to increase cross-sectional nasal valve area and enhance air flow through the nose. When worn, the device applies a gentle outward pull to the outside walls of the nasal vestibule that helps prevent collapse of the lateral walls of the nasal vestibule. The device appears similar to an adhesive bandage with two polyester springs attached lengthwise to the nonadhesive side of the backing material. The form of the external nose varies significantly between racial groups and some comment is possible about the form of the nasal valve, i.e., Caucasian noses are leptorrhine, Negroid noses platyrrhine, and Oriental noses intermediate. A question exists about whether Breathe Right is as effective in Japanese as in Caucasians against nasal obstruction. We measured the effect by acoustic rhinometry in 35 healthy subjects. We measured 2 cross-sectional areas the I-notch and C-notch-and 3 nasal volumes-V(0-1), from 0 to 1 cm corresponding to the I-notch, V(1-3), from 1 to 3cm corresponding to the C notch, and V (3-6), from 3 to 6cm. We studied the influence of the nasal cycle on the effect of Breathe Right in the nasal cavity. We evaluated subjects in a stable sitting position before applying Breathe Right, after using 5 minutes, and in a free interval of 25 minutes following. The experiment was repeated 10 times. We found that Breathe Right increases the nasal volume especially in V(1-3) and the cross sectional area especially in the C-notch. Interestingly, the change in the C notch decreased gradually with time. One possible mechanism may be the change in compliance between the skin under the Breathe Right and the nasal mucous membrane. The nasal cycle did not significantly influence the effect of Breathe Right. PMID- 16359005 TI - [Image-guided endoscopic sinus surgery: a comparison of two navigation systems]. AB - We evaluated the effectiveness of two navigation systems with optical tracking in endoscopic sinus surgery (ESS). The Signa SP/i Intraoperative navigation system (General Electric Co., Intraoperative NS) is advantageous in acquiring both real time and high-resolution images during surgery, compared to conventional image guided navigation (Stealth Station TREON, Medtronic Inc., IGNS) that rely entirely on preoperative three-dimensional images. We studied the following in 14 patients treated with intraoperative NS and 19 treated with IGNS: 1) additional time for navigation system implementation, 2) available instrumentations in ESS, and 3) navigation system accuracy. Navigation systems required additional time to prepare ESS. The time lapse from admittance to the operating room to ESS onset was measured in patients under both systems and controls undergoing ESS without any image guidance. Preparation of the intraoperative NS required an additional 52 min and IGNS required 17 min compared to the control group. Based on operative instruments, the intraoperative NS has some limitations arising from the application of a high magnetic field. Surgical instruments must not be attracted by the magnetic field in the operating room. So those used in our study were thoroughly examined and some remanufactured using MR safe materials. All instruments in ESS could be used in surgical guidance during surgery in the IGNS. Anatomic landmarks were accurately visualized using intraoperative NS and IGNS. Intraoperative NS renews the image during surgery, so surgeons could confirm the surgical outcome during ESS. Since the average distant error in both systems was between 1 mm and 2.5 mm, we confirmed that accuracy obtained with both navigation system was suitable for ESS completion. The intraoperative NS renewed the image during navigation. In conclusion, both navigation systems are sufficient for accurate image navigation in ESS, but navigation systems must be selected based on the individual case. PMID- 16359006 TI - [Comparison of audiological performance bewteen bone-anchored and conventional hearing aids]. AB - The bone-anchored hearing aid (BAHA) is an implantable bone-conduction device that vibrates the skull directly via a surgically implanted titanium screw behind the ear. The BAHA has advantages for patients with aural atresia or chronic ear drainage, who cannot wear air-conduction hearing aids. We compared the function of BAHA to conventional bone and air-conduction hearing aids based on functional gain and speech discrimination tests in quiet and noisy environments in 2 patients with chronic ear problems. All expressed a clear preference for the BAHA over conventional bone-conduction hearing aids. The BAHA and air-conduction hearing aid provided similar audiological performance when the functional gain of each hearing aid coincided. As the air-bone gap widens, however, audiological performance with the BAHA showed an evident preference, indicating that the width of the air-bone gap is of some help in BAHA preference compared to air-conduction hearing aids. PMID- 16359007 TI - [Early and long-term results of valve replacement in patients on chronic hemodialysis]. AB - Cardiac valve replacement carries a high-risk in hemodialysis patients and has been questioned for its results. The purpose of this study is to focus on the early and long-term results in our institution. We retrospectively analyzed the data from 41 hemodialysis patients who underwent valve replacement in our unit from July 1978 to March 2004. Bioprosthetic valves were used in 5 patients (12.2%). The overall operative mortality (30 days) was 17%. The in-hospital death rate was 22%. Late survival rate was 68% at 2 years, 54% at 3 years, 49% at 5 years, and 27% at 10 years for all patients. The incidence of major bleeding and stroke was 41%. PMID- 16359008 TI - [Double outlet right ventricle and pulmonary atresia with a birth weight of 1092 g]. AB - A premature infant with double outlet right ventricle and pulmonary atresia with a birth weight of 1092 g is reported. He underwent right modified Blalock-Taussig (RMBT) shunt with an expand-polytetrafluoroethylene (ePTFE) tube of 3.0 mm in diameter between the right subclavian artery and the right pulmonary artery through right thoracotomy. Eleven days later, he had to undergo central shunt between the innominate artery and the main pulmonary trunk due to poor pulmonary blood flow. Soon after the central shunt, severe heart failure occurred due to excessive pulmonary blood flow. RMBT division was performed immediately. He finally attained definitive repair at 17 months of age. Postoperative course was uneventful and he was discharged on the 17th postoperative day. PMID- 16359009 TI - [Chronic tuberculous methicillin-resistant Staphylococcus aureus (MRSA) empyema with bronchopleural fistulae treated by open window thoracostomy followed by thoracoplasty and latissimus dorsi muscle transposition]. AB - We describe a case of chronic tuberculous methicillin-resistant Staphylococcus aureus (MRSA) empyema with bronchopleural fistulae successfully treated by open window thoracostomy followed by thoracoplasty and transposition of the latissimus dorsi muscle. A 69-year old man with a history of artificial pneumothorax for pulmonary tuberculosis was referred to our hospital with fever and purulent bloody sputum. He was diagnosed as having right chronic tuberculous empyema with bronchopleural fistulae. Immediate tube thoracostomy markedly relieved symptoms except for low-grade fever. Sputum and empyema cavity cultures were repeatedly positive for MRSA. Open window thoracostomy (5th to 7th ribs resection) was performed to control the infection. The empyema cavity was cleaned with no residual calcified pleura. His condition gradually improved and he underwent thoracoplasty and transposition of the latissimus dorsi muscle 22 months after the initial surgery. He was discharged 25 days postoperatively in good condition. Seventeen months after the curative surgery, he remains well with no evidence of recurrence. A two-stage operation, open window thoracostomy to control infection followed by thoracoplasty and transposition of the latissimus dorsi muscle, is useful in cases of chronic tuberculous MRSA empyema with bronchopleural fistulae. PMID- 16359010 TI - [Jatene procedure with arch repair; usefulness of rapid two-stage repair]. AB - Four cases of Jatene procedure with arch repair were performed since 2000 in our department. Two-stage repair was used in all cases and the extended end-to-end anastomosis and pulmonary artery banding (PAB) were performed in 3 cases as the initial repair. In a recent case of TGA type I with coarctation of the aorta (CoA), only subclavian flap aortoplasty was performed when he was 6-day-old and patent ductus arteriosus (PDA) was preserved and rapid two-stage Jatene procedure was performed when he was 8-day-old. There was no hospital or late death, or reoperation. The results of the two-stage Jatene procedure with arch repair was good and safe. Rapid two-stage repair was thought to be a useful choice especially for TGA type I with arch anomaly. PMID- 16359011 TI - [Mitral valve repair for commissural prolapse due to infective endocarditis]. AB - A 65-year-old man and a 51-year-old man underwent mitral valve repair for commissural prolapse due to infective endocarditis. On the occasion of repairing, folding plasty technique was employed to avoid relatively large annular plication after leaflet resection. Postoperative echocardiography showed no residual regurgitation and sufficient orifice area of the mitral valve. Folding plasty technique appeared to be simple and useful for repairing commissural prolapse due to infective endocarditis. PMID- 16359012 TI - [Prophylactic treatment of oral sildenafil citrate for pulmonary hypertension in an infant treated with inhaled nitric oxide after open-heart surgery]. AB - A 3-month-old boy with coarctation of the aorta (CoA), ventricular septal defect (VSD), atrial septal defect, and severe pulmonary hypertension (PH) underwent one stage repair consisting of patch closure of VSD and coarctation repair. Inhalation of nitric oxide (iNO) was commenced to treat residual severe PH on the day of the operation. Oral sildenafil citrate was commenced on the day 1 and iNO was gradually weaned off on the day 3. There was no "rebound", severe increase in pulmonary artery pressure, which commonly occurs after discontinuation of iNO. Then the patient was extubated without any difficulties or recurrent PH. The oral sildenafil citrate therapy was ceased on the day 8. Prophylactic use of oral sildenafil citrate for PH might be an useful alternative to shorten the duration of iNO therapy and intensive care unit (ICU) stay in the selected patients after congenital open heart surgery. PMID- 16359013 TI - [Aortic valve replacement for the patient with Osler-Rendu-Weber disease]. AB - Osler Rendu-Weber (O-R W) disease (hereditary haemorrhagic telangiectasia) is characterized by an anormality of the peripheral blood vessels which often causes recurrent epistaxis. We presented a case of aortic valve replacement for the patient with O-R-W disease. A 75-year-old woman diagnosed as O-R-W disease with chest pain and dyspnea was admitted. Preoperative transthoratic echocardiography showed severe aortic valve stenosis (AS) with a pressure gradient (from left ventricle to ascending aorta) of 72.5 mmHg. Urgent aortic valve replacement (AVR) was performed after preoperative hormonal (estrogen-progesterone) therapy, that might be effective for bleeding from vascular ectasia. No hemorrhagic tendency was recognized and the postoperative course was uneventful. PMID- 16359014 TI - [Staged surgical palliations for hypoplastic left heart syndrome with poor preoperative conditions]. AB - A 2.8 kg-female newborn diagnosed as hypoplastic left heart syndrome was transferred to our hospital after resuscitation against systemic circulatory failure due to closure of the ductus arteriosus. Her systemic circulation was stabilized after alprostadil (lipo-PGE1) and inotorpic agent infusion along with mechanical ventilation support. On admission, signs of a pulmonary high-flow and low systemic perfusion were evident. Impaired coagulation aspect was gradually noted. Because a surgical palliation with cardiopulmonary bypass was considered to be at high-risk, a bilateral pulmonary artery banding was indicated. Through a median sternotomy, the right and left pulmonary arteries were individually banded. The patient was continued on the same ventilation strategy. Cardiac, hepatic and renal dysfunctions were improved over next few days and the patient weaned from ventilatory support. She underwent combined Norwood stage I and II repair at 4 months of age with weight of 4.8 kg. Postoperative course has been uneventful and the patient is now followed up in preparation for Fontan operation. PMID- 16359015 TI - [Castleman disease of the inter-lobar lymph node origin]. AB - We report a case of Castleman disease which originated from the inter-lobar lymph node, with a review of literatures. A 19-year-old woman complaining of cough was pointed out to have an abnormal shadow in the left lung field on chest X-ray. Chest computed tomography (CT) and magnetic resonance imaging (MRI) with enhancement revealed a homogeneous mass lesion at the left inter-lobar portion of the lung. Bronchoscopic findings demonstrated mucosal telangiectasis of the left lower bronchus. We performed the usual axillary thoracotomy and succeeded in extirpation of the tumor without large amount of bleeding. The tumor was elastic and hard, and 70 x 55 x 45 mm in size. Her postoperative course was uneventful and she was discharged on the 12th postoperative day. PMID- 16359016 TI - [Lobar torsion after pulmonary resection for lung cancer]. AB - We reviewed our experience on postoperative lobar torsion. From January 1994 to December 2003, 1002 patients underwent lobectomy for lung cancer. Two (0.2%) patients with postoperative lobar torsion required surgical reintervention. The first case was that of a 79-year-old man who underwent left lower lobectomy for pulmonary adenocarcinoma. Based on the postoperative course, lobar torsion was highly suspected with progressive respiratory dysfunction and the chest X-ray showed complete opacification of the residual lobe. Rethoracotomy was performed on postoperative day 4. The left upper lobe was rotated clockwise, and completion pneumonectomy was carried out. The patient died 16 days after the second surgery because of respiratory failure due to severe pneumonia. The second case was that of a 24-year-old man with a diagnosis of metastatic lung cancer in the right upper lobe arising from pharyngeal cancer. The patient underwent right upper lobectomy and developed hemoptysis and persistent high fever. A chest computed tomography (CT) and bronchoscopy findings revealed lobar torsion of the middle lobe, and a reoperation was performed. The middle lobe was resected and the patient was discharged 8 days after the rethoracotomy. Pulmonary lobar torsion poses a difficult diagnostic dilemma in the early postoperative period after the pulmonary resection. Since late reoperation for postoperative lobar torsion sometimes results in poor prognosis, careful observation with bronchial fiberscopy as well as chest radiography is necessary for accurate diagnosis. Rethoracotomy should be carried out without any delay in cases of lobar torsion following pulmonary resection. PMID- 16359017 TI - [Reoperative off-pump coronary artery bypass via left thoracotomy with porcelain aorta; left axillary artery to circumflex artery bypass; report of a case]. AB - A 62-year-old man who underwent coronary artery bypass grafting (CABG) [left internal thoracic artery (LITA)-left anterior descending (LAD), saphenous vein graft (SVG) right coronary artery (RCA)] 13 years previously developed angina pectoris and congestive heart failure because of occlusion of SVG and native vessels. Coronary angiography (CAG) revealed that inflow to the coronary artery remained only from LITA. Repeat off-pump CABG (OPCAB) with SVG to the circumflex artery via left thoracotomy was performed. The proximal end of SVG was anastomosed to the left axillary artery because of the porcelain aorta and the patent LITA graft. The patient developed no complications and was discharged from hospital on postoperative day 21. OPCAB for circumflex artery by left thoracotomy is an effective and safe approach in redo CABG, particularly in instances of patent LITA. PMID- 16359018 TI - [Cardiac papillary fibroelastomas: report of 2 cases]. AB - We report 2 cases of cardiac papillary fibroelastomas in adults. Case 1: A 61 year-old man was admitted because echocardiography showed a 1 cm pedunculated papillary tumor in the left atrium. In an operation, it was located in the left atrium near the mitral valve and was resected along with a 5 mm margin of endocardium. Case 2: A 60-year-old woman had a 1 cm mobile tumor in the right ventricle near the tricuspid valve located by echocardiography in a preoperative examination of a ventricular septal defect. In an operation, a pedunculated tumor located in the right ventricle was resected. In these 2 cases, histopathology showed the tumor to be a papillary fibroelastoma. Almost all cardiac papillary fibroelastoma are closely related to the cardiac valve, but in these cases, the tumors were located in the left atrium, and the right ventricle, respectively, which is quite rare. PMID- 16359019 TI - [Mature teratoma combined with schizophrenia: report of a case]. AB - A 20-year-old woman was admitted to another hospital due to schizophrenia in July 2003. The patient felt chest pain and palpitation in August, and she was referred to our hospital. Chest computed tomography (CT) showed a mass in the left thoracic cavity and a pleural/pericardial effusion. Since general condition did not improved in spite of aggressive supportive treatment, surgical treatment was chosen. An operation was performed via median sternotomy in September. The tumor was found to have adhered firmly to the surrounding organs. Pericardial fenestration was performed; then the tumor was resected with the left phrenic nerve due to tight adhesion. The patient received respiratory support in the intensive care unit for 5 days after surgery, uneventfully. Twenty-three months after surgery, she is surviving and free from schizophrenic symptoms without medication. PMID- 16359020 TI - [Bronchial artery embolization for treatment of mediastinal hemorrhage after pulmonary resection: report of a case]. AB - We report a rare case of mediastinal hemorrhage after pulmonary resection. A 64 year-old woman with hypersensitivity pneumonitis was diagnosed as adenocarcinoma of the lung by bronchoscopical examination. Left lower lobectomy and mediastinal lymph node dissection were performed. Sudden chest pain and dry cough developed 14 days after the operation. Her diastolic pressure rose transiently but electrocardiogram remained normal. Chest X-ray showed widening of the mediastinum and enhanced chest computed tomography (CT) showed extravasation of the contrast media just under the bifurcation of the trachea. Multi projection volume reconstruction revealed mediastinal hemorrhage from the bronchial artery. The chest pain disappeared after a successful bronchial artery embolization and the patient discharged 21 days later. Hemorrhage after pulmonary resection is a common complication, but no previous report has described mediastinal hemorrhage occurring 2 weeks after the operation. In a similar case, bronchial artery embolization is a reliable and minimally invasive therapy for mediastinal hemorrhage. PMID- 16359021 TI - [Thoracolithiasis detected during video-assisted thoracoscopic surgery for spontaneous pneumothorax: report of a case]. AB - A 32-year-old male was admitted to our hospital for treatment of right recurrent spontaneous pneumothorax. Chest X-ray showed right pneumothorax but chest X-ray and computed tomography (CT) examinations did not demonstrate intrathoracic abnormal mass shadow or abnormal calcifications. We performed video-assisted thoracoscopic surgery for spontaneous pneumothorax. At surgery, an egg-shaped milky white free mass was found in the thoracic cavity, and easily removed. This substance was hard and smooth on the surface and measured 12 x 9 mm. Histologically this mass was composed of fatty necrosis surrounded by hyalinized connective tissue. These findings led to a diagnosis of thoracolithiasis. Thoracolithiasis without any history of chest trauma or intervention is pathologically rare, with only 15 cases including our case in Japan, having been reported in the literature to date. PMID- 16359022 TI - [Non-Hodgkin malignant lymphoma of rib origin: report of a case]. AB - A 69-year-old man with left chest and back pain was found to have an osteolytic mass (4.2 x 3.8 cm) in the left 8th rib by chest X-ray and computed tomography (CT) in August 2003. There were no abnormal findings in the abdomen, lung, mediastinum or bone except the left 8th rib. Although the spontaneous disappearance of pleural effusion and the elevated CRP suggested the possibility of myelitis, the malignancy of the rib could not be ruled out, and the surgery was performed in September 2003. The mass was resected en bloc together with the involved 8th rib. The histological diagnosis was primary non-Hodgkin lymphoma (diffuse, medium-sized to large B-cell lymphoma). PMID- 16359023 TI - [Lung cancer with polyarthropathy due to parvovirus B19: report of a case]. AB - We reported a case of lung cancer with polyarthropathy induced by human parvovirus B19. A 60-year-old female with lung cancer was admitted to our hospital with polyarthropathy similar to rheumatoid arthritis. Although the abnormal accumulation of 99mTc-MDP in the bone scan film and rheumatoid factor positive were detected before operation, we diagnosed polyarthropathy induced by parvovirus B19 through the detection of parvovirus B19 IgM and B19 DNA. The joint pain gradually improved after surgical treatment for lung cancer. PMID- 16359024 TI - [Pleural lipoma: report of a case]. AB - Although lipomas are the most common form of the benign neoplasm, occurrence within the thoracic cage is uncommon, and lipomas originating from the pleura are very rare. We report an unusual case of a 63-year-old male whose serial chest X ray demonstrated an abnormal shadow of which size was increasing. A chest wall tumor was suspected based on the findings of computed tomography (CT) of the thorax. The tumor was resected by video-assisted thoracoscopic surgery and the patient remains well with no recurrence 4 years postoperatively. The resected tumor was a yellowish pleural mass and the pathological diagnosis was a benign lipoma. It is possible to make a diagnosis of lipoma preoperatively by CT and magnetic resonance imaging (MRI), however, the correct differential diagnosis of lipoma and liposarcoma is imperfect only by radiological findings necessitating surgical removal if possible. However, in view of the imperfect differentiation between lipoma and liposarcoma, surgical treatment should be considered. PMID- 16359025 TI - [Thoracoscopic clipping of the thooracic duct for chylothorax following esophagectomy: report of a case]. AB - Thoracoscopic clipping of the thoracic duct was successfully performed for the treatment of postoperative chylothorax. Chylothorax occurred in a 67-year-old man following an esophagectomy for esophageal cancer. Following unsuccessful conservative therapy for 3 weeks, we performed thoracoscopic surgery to examine the thoracic duct and found a leaking point of chylous fluid. The thoracic duct was successfully clipped resulting in complete elimination of the effusion immediately after surgery. Generally, chylothorax complicated by an esophagectomy has been managed by medical treatment first, followed by surgical intervention in case of uncontrollable pleural effusion. We think you should try this method at first in case chylothorax was able to be treated with not thoracotomy but thoracoscopic surgery: minimal invasiveness. PMID- 16359026 TI - [Application of transcranial magnetic MEP to epilepsy surgery: the decision of operative course and the relationship with seizure outcome]. PMID- 16359027 TI - [Microanatomy of the temporal bone]. PMID- 16359028 TI - [Study on hemosiderin deposition after intracerebral hemorrhage]. AB - BACKGROUND AND PURPOSE: Hemosiderin degenerated from intracerebral hematoma (ICH) can be visualized as low intensity on gradient-echo T2*-weighted (T2*-w) MRI, which is sensitive for hemosiderin. We studied the factors associated with the hemosiderin deposition on T2*-w MRI after ICH. METHODS: We analyzed T2*-w MRIs in 72 outpatients with past ICH (46 males, 26 females, 28-89 (60.0 +/- 9.8) years old) consecutively came to our hospital. The odds ratio (OR) for the apparent thickness > or = 2.5 mm of low intensity surrounding ICH cavity was estimated, using the time after the onset of ICH (17 weeks - 26 years), and other factors. RESULTS: All of old ICHs were visible as low intensities on T2*-w MRI. Multivariate logistic regression analysis revealed that an elevated ratio of the apparent thickness > or = 2.5 mm of hemosiderin was found for the patients with time > or =5 years between T2*-w MRI and the onset (odds ratio (OR): 0.24, 95% confidence interval (CI): 0.06-0.99), intraventricular or subarachnoid hemorrhage related to ICH (OR: 0.16, 95% CI: 0.03-0.77), and the diameter of ICH > or =2 cm (OR: 33.7, 95% CI: 4.6-245). CONCLUSION: Though small sample size limited the power of analyses, our findings suggest that the amount of hemosiderin deposition after ICH may be associated with the time after the onset, intraventricular or subarachnoid hemorrhage related to ICH, and the diameter of ICH. PMID- 16359029 TI - [An elderly case of craniopharyngioma: treatment by limited surgery and radiation therapy]. AB - Craniopharyngioma accounts for approximately 3.5% of all intracranial neoplasm and may occur at any age. However craniopharyngioma in the elderly was rare. We report a rare case of craniopharyngioma in an elderly patient, who was treated by radical surgery successfully. An 80-year-old male was admitted to our hospital with a history of visual disturbance and bitemporal hemianopsia. CT scan of the brain revealed a hypodense cystic mass with calcification in the suprasellar area. MRI demonstrated the optic chiasm was displaced superiorly. Her symptom deteriorated day short time of her hospitalized life. We decided to removal tumor radically because of saving her visual function. Subtotal removal of the suprasellar lesion was performed, followed by radiotherapy to the residual lesion. A visual impairment was improved after surgery. No recurrences had been detected on MRI eighteen months after the initial operation. Based on previous report and the clinical course of our case, radical surgery is sometimes needed for saving visual function in spite of elderly patient. However, in this case, sparing hypothalamic and pituitary function is essential for good prognosis. PMID- 16359030 TI - [A case of the lateral ventricle lesion mimicking arachnoid cyst treated by neuroendoscopic surgery]. AB - A 12-year-old female suffered from intermittent headache with nausea. Neuroradiologic studies demonstrated a cystic lesion in the right trigone with content like cerebrospinal fluid. The right inferior horn was isolated and increased in size. Eodoscopic fenestration was attempted using a flexible fiberscope via the inferior horn. Postoperative magnetic resonance image revealed shrinkage of the cyst and reduced size of the right inferior horn. Intermittent headache dissapeared. Neuroendoscopic treatment is a less invasive and effective therapy for intraventricular arachnoid cyst as well that in the other intracranial lesion. PMID- 16359031 TI - [A case of intracranial infectious granuloma extending into subcutaneous and subdural space]. AB - A case of intracranial infectious granuloma extending into subcutaneous and subdural space mimicking malignant bone tumor was reported. A 49-year-old male was admitted to our service with complaint of left frontal subcutaneous and bone tumor. Neurological examination demonstrated mild disorientation only except for pre-existeng Schizophrenia. CT scan and Gadolinium-enhanced MRI revealed left frontal intraosseous and subdural mass accompanied with remarkable edema. Left frontal bone was destructed by this mass. However, inflammatory sign nor malignant tumor could be observed and left frontal craniotomy and removal of the tumor was performed. The tumor was solid, elastic hard and existing from intra osseous, epidural and subdural space compressing the cortex. No abscesss cavity could be found. Histological examination revealed infectious granuloma accompanied by remarkable neutrophil infiltration only. The patient showed good post-operative course, and returned to previous condition, pre-existeng Schizophrenia only. Intracranial infectious granuloma extending into subcutaneous and subdural space destructing the bone was rare condition. From literature, similar cases could be found in granuloma arising from osteomyelitis. In this case, destructed bone beneath the epi- and subdural tumor support this route of infection. PMID- 16359032 TI - [Neuroendoscopic placement of the reservoir in an elderly patient with recurrenced craniopharyngioma: case report]. AB - We report a case of recurrent craniopharyngioma in the third ventricle with obstructive hydrocephalus, which was successfully treated by placement of the Ommaya reservoir by neuroendoscopic procedure. A 72-year-old male with disorientation and gait disturbance was admitted to our hospital. He had been suffering chronic heart failure and arrhythmia due to mitral valve insufficiency, and panhypopituitarism after the first craniotomy for craniopharyngioma. MRI demonstrated obstructive hydrocephalus at the foramen of Monro by the cystic tumor. Cyst decompression and placement of Ommaya reservoir were successfully performed in local anesthesia. Postoperatively, his disorientation and gait disturbance were improvement, and no chemical meningitis developed. Neuroendoscopic management for cystic craniopharyngioma with obstructive hydrocephalus was effective procedure for elderly patient with systemic risk factor. PMID- 16359033 TI - [Symptomatic Rathke's cleft cyst mimicking suprasellar arachnoid cyst]. AB - The authors report an atypical case of symptomatic entirely suprasellar Rathke's cleft cyst mimicking suprasellar arachnoid cyst. A 55-year-old male was introduced to our hospital complaining of bitemporal hemianopsia. CT and MRI demonstrated a cystic mass located entirely in the suprasellar cistern and to compress the optic nerve and mammillary body. The cystic wall was not enhanced in MRI. CT cisternography showed the suprasellar non-communicating cyst with cistern. The hormonal function was slightly disturbed by the pituitary compression. Under the diagnosis of suprasellar arachnoid cyst, a left front temporal craniotomy was performed to resect the suprasellar mass. The surgical specimen consisted ciliated epithelium and was diagnosed Rathke's cleft cyst. After operation, he recovered completely free. PMID- 16359034 TI - [Two surgically treated cases with dural arteriovenous malformation in the anterior cranial fossa]. AB - Dural arteriovenous fistulae(dAVF) in the anterior cranial fossa comprise about 6% of all dAVFs, and is usually detected after intracranial hemorrhage. However, non-hemorrhagic symptoms are uncommon. We encountered two patients with hemorrhagic and non-hemorrhagic dAVF in the anterior cranial fossa. Both cases were successfully treated using craniotomy and histopathological examination was performed. A 71-year-old male experienced sudden onset of nausea and headache, followed by progression of left hemiparesis and disturbance of consciousness due to intracerebral and subdural hematoma in 2005. Intracerebral venous aneurysm adjacent to the intracerebral hematoma originated from the draining cortical red vein was identified and successfully treated by operation. Histopathological examination revealed disappearance of the internal elastic lamina and scant muscle tissue in the venous aneurysmal wall originating from draining cortical vein. A 69-year-old male suffered sudden onset of mild headache, followed by progression of bruits in 2005. MRI suggested dAVF in the left anterior cranial fossa. 123I-IMP SPECT revealed an area of low perfusion in the left frontotemporal region. Histopathological examination after surgical treatment revealed thickening of the intima and an indistinct lamina elastica interna, indicating progressive degenerative changes of the arterialized vein likely to result in hemorrhage. Surgical obliteration of fistulae is reasonable even in the dAVF of the anterior cranial fossa with ischemic onset. PMID- 16359035 TI - [A case of atypical meningioma with Lhermitte-Duclos disease]. AB - Lhermitte-Duclos disease is known as an uncommon disease that characterized by a slowly progressive tumor of the cerebellar hemisphere. We present a case of atypical meningioma with Lhermitte-Duclos disease. A 57-year-old female was admitted to our hospital after presenting with general convulsion. CT scan on arrival showed a large mass of the left frontal region and slightly high-density linear lesion in the right cerebellar hemisphere. MRI also revealed well-enhanced mass in the left frontal region and parallel linear striation in the right cerebellar hemisphere. But, the patient had neither cerebellar dysfunction nor signs of increased intracranial pressure. The patient had angioma of the left breast and bilateral benign struma, no typical manifestation of Cowden syndrome. Removal of the frontal tumor caused the convulsion was subsequently performed. The pathological examination was atypical meningioma. After the operation, radiation therapy was not done because of the total removal of tumor and intension on patient side. Fortunately, the patient had no further adverse neurological events postoperatively. However, we should continuously take account to not only the recurrence of meningioma but also the enlargement of the cerebellar lesion and the complication of malignant tumors in whole body. PMID- 16359036 TI - [Management of diagnosis procedure combination (DPC) in neurosurgical clinic--new insurance payment system introduced into the hospital in Japan]. PMID- 16359037 TI - [Surveillance of acute hepatitis B with regard to vaccination]. PMID- 16359038 TI - [Shortage of nurses in the Western World]. PMID- 16359039 TI - [Sterilization in the hospital environment]. PMID- 16359040 TI - [Preparation of instruments in the operating room]. PMID- 16359041 TI - [Foot ulcers]. PMID- 16359042 TI - [Co-infection with HIV-hepatitis C virus]. PMID- 16359043 TI - [Consultation at Baudelire: the hospital environment at the center of attention]. PMID- 16359044 TI - [Equipment and procedures of enteral nutrition]. PMID- 16359045 TI - [Oseltamivir]. PMID- 16359046 TI - [Prenatal echography: valid or not valid?]. PMID- 16359047 TI - Editorial: An 'actual' problem: Another issue of apples and oranges. PMID- 16359048 TI - Mitral valve surgery using the classical 'heartport' technique. AB - BACKGROUND AND AIM OF THE STUDY: Mitral surgery in selected patients using femoral cannulation, percutaneous retrograde cardioplegia and endoballoon aortic occlusion with a 4-5 cm thoracotomy is felt to carry a higher operative risk than sternotomy with conventional cannulation. Herein, the authors compared their experience of the first 117 'Heartport' (HP) patients with a computer-matched group of sternotomy approach surgical patients (SP) to assess operative risk and 30-day outcome. METHODS: Data were extracted from the authors' STS certified, audited database on 117 patients based on an intention to treat. Between December 1997 and December 2004, a total of 92 isolated mitral valve (MV) repair (HP MVRpr) and 25 isolated MV replacement (HP-MVR) procedures was conducted using Heartport. The patients were matched 1:1 (by age +/-7 years, cerebrovascular disease, inotrope use, and ejection fraction +/- 5%) to a control SP group. Operative and 30-day outcomes were measured. RESULTS: No parameter showed any significant difference in 30-day outcome between the HP and SP groups, except for an increase in cross-clamp and perfusion times. Two patients in the HP-MVRpr group required conversion to sternotomy for repair of coronary sinus perforation, and one patient was repaired without conversion. Surgery in one HP-MVRpr patient was aborted due to limited aortic dissection, but successful repair was carried out later with a conventional approach. No patient required conversion to sternotomy for improved exposure of the operative site. CONCLUSION: Despite a longer cross-clamp time, 'classical' HP MV surgery can be performed with no increased risk compared to conventional MV surgery. Catheter and endoballoon complications were rare even in the early experience, and conversion to sternotomy was unusual and safely performed with this approach. PMID- 16359049 TI - Mitral valve reconstruction: experience related to early and late mortality and reoperation. AB - BACKGROUND AND AIM OF THE STUDY: The present authors' experience with mitral valve reconstruction was reviewed to determine the influence of anatomical and functional abnormalities on mortality and reoperation. METHODS: Between 1991 and 2001, a total of 397 patients (mean age 59.0 +/- 13.4 years) underwent mitral valve reconstruction at the authors' institution. Patients were grouped according to the Carpentier functional classification as follows: type I, n = 88 (mean age 57.4 years; range: 24-81 years); type II, n = 169 (mean age 59.8 years; range: 16 81 years); type IIIa, n = 44 (mean age 52.8 years; range: 22-78 years); type IIIb, n = 85 (mean age 65.1 years; range: 33-82 years); and congenital (C), n = 11 (mean age 36.2 years; range: 18-65 years). RESULTS: The total follow up was 1,485.8 patient-years (pt-yr). Early mortality was: type I, 3.4% (n = 3); type II, 0; type IIIa; 2.3% (n = 1); type IIIb, 9.4% (n = 8); and C, 0 (p = 0.0028, IIIb > II and C). The late mortality was: type I, 2.1%/pt-yr (n = 6); type II, 1.1%/pt-yr (n = 7); type IIIa, 1.8%/pt-yr (n = 4); type IIIb, 4.3%/pt-yr (n = 12); and C, 1.7%/pt-yr (n = 1) (p = 0.0035 IIIb > II). The overall survival at 10 years was 79.6 +/- 4.2% (p <0.001, II > IIIb; p = 0.029, I > IIIb; p = 0.046, II > I). The reoperations were: type I, 1.4%/pt-yr (n = 4); type II, 1.9%/pt-yr (n = 12); type IIIa, 0.9%/pt-yr (n = 2); type IIIb, 0.4%/pt-yr (n = 1); and C, 0 (p = 0.0435 II > IIIb). Among the 19 reoperations there were 17 replacements, one re repair, and one annuloplasty. The failures were predominantly type II, posterior (n = 5) and anterior (n = 5), or a combination (n = 2). The overall freedom from reoperation was 92.0 +/- 2.2% (p = NS between groups). CONCLUSION: Mitral valve reconstruction can be performed in all four functional categories with generally satisfactory results, except possibly for functional ischemic disease. The results of surgery for degenerative disease afford the opportunity for early surgical management, if proven techniques are adhered to in order to minimize the risk of reoperation. The repair of ischemic disease due primarily to ventricular dysfunction is in a state of evolution. PMID- 16359050 TI - Quality of life after mitral valve repair. AB - BACKGROUND AND AIM OF THE STUDY: Mitral valve repair for degenerative and ischemic mitral valve regurgitation has been shown to be a durable procedure. The study aim was to evaluate the quality of life of patients who had undergone mitral valve repair, and to compare it to that of an age- and gender-adjusted Finnish general population. METHODS: Among 130 late survivors after mitral valve repair, 109 (83.8%) answered the RAND-36 Health Survey questionnaire; these patients form the basis of the present study. RESULTS: The Wilcoxon test showed significantly higher mental health (p = 0.04) and pain scores (p = 0.015) and a lower role functioning/physical score (p = 0.008) in the study group. The scores of the other RAND-36 Health Survey variables of the study group were similar to those of the age- and gender-adjusted general population. The mean total score for the study group was 512 (median 532, IQR 360-678), compared to 522 (median 538, IQR 468-549) in the general population (p = 0.72) (only 95 patients were included in the analysis due to isolated missing scores). CONCLUSION: The quality of life of long-term survivors after mitral valve repair, as assessed by the RAND 36 Health Survey, is similar to that of an age- and gender-adjusted general Finnish population. PMID- 16359051 TI - Balloon mitral commissurotomy for patients with mitral stenosis in atrial fibrillation: ten-year clinical and echocardiographic actuarial results. AB - BACKGROUND AND AIM OF THE STUDY: Controversy persists as to whether atrial fibrillation (AF) has a direct negative effect on the outcome of percutaneous balloon mitral commissurotomy (PBMC). The study aim was to assess the effect of AF on immediate and 10-year clinical and echocardiographic actuarial results of patients with rheumatic mitral stenosis undergoing PBMC. METHODS: A total of 195 consecutive patients with AF (group 1) was analyzed and compared with 195 patients in sinus rhythm (group 2), matched for the severity of mitral valve morphological changes. RESULTS: Group 1 patients were older (43.8 + 12 versus 30.5 +/- 12.7 years; p <0.001), and a greater proportion had grade 1 mitral regurgitation (25.1 versus 9.7%; p <0.0001). The procedural success was 89.8% in group 1 and 92.3% in group 2 (p = NS), but group 1 patients had a smaller mitral valve area (2.1 +/- 0.4 versus 2.3 +/- 0.4 cm2; p <0.0001). Patients in AF had a lower 10-year survival (91.4 versus 99.4%; p = 0.018), a lower 10-year event-free survival (60.3 versus 70%; p = 0.02), and a lower 10-year freedom from restenosis (40 versus 66%; p = 0.048). AF was an independent predictor of overall mortality (OR = 10.79; p <0.033) and of combined events (death, redo PBMC or mitral valve surgery; OR = 1.95; p <0.012), and was a univariate predictor of restenosis (p = 0.048). CONCLUSION: Patients with AF have good immediate results, but poorer long term outcome after PBMC. AF a marker of worse prognosis in patients with mitral stenosis as it is in the general population. PMID- 16359052 TI - N-terminal proBNP plasma levels correlate with severity of mitral stenosis. AB - BACKGROUND AND AIM OF THE STUDY: Brain natriuretic peptide (BNP), a neurohormone, is secreted predominantly from the ventricular myocardium. Studies investigating BNP secretion in diseases affecting the right side of the heart are scarce. The relationship between N-terminal proBNP (NT-proBNP) and echocardiographic and clinical findings was studied in cases with isolated moderate to severe rheumatic mitral stenosis (MS), and in patients with previous mitral valve replacement (MVR). METHODS: Thirty-two patients with MS (mean age 41.2 +/- 5.7 years), 20 with MVR (mean age 46.0 +/- 4.6 years) and 30 healthy individuals (mean age 40.3 +/- 4.9 years) were included in the study. In addition to NT-proBNP measurements, detailed transthoracic echocardiography was performed in all patients and healthy subjects. RESULTS: Plasma levels of NT-proBNP were significantly higher in patients with MS than in those with MVR or in controls (99.8 +/- 12.7 versus 74.7 +/- 6.9 and 48.5 +/- 10.5 pg/ml, respectively; p <0.0001 all groups). NT-proBNP levels showed a significantly greater increase in severe MS than in moderate MS (109.8 +/- 5.6 versus 88.3 +/- 7.6 pg/ml, p <0.0001). NT-proBNP levels also were higher in MVR patients than in controls (74.7 +/- 6.9 versus 48.5 +/- 10.5 pg/ml; p <0.0001). Although NT-proBNP levels did not correlate with left ventricular ejection fraction (LVEF) in patients with MS (r = -0.33; p >0.05), there was a positive correlation with pulmonary artery pressure (r = 0.87; p <0.001) and a negative correlation with mitral valve area (MVA) (r = -0.89; p <0.0001). However, multivariate analysis identified only MVA as an independent correlate of NT-proBNP (beta = -0.47; p = 0.02). CONCLUSION: In patients with rheumatic MS, NT proBNP levels correlated positively with MS severity. Moreover, NT-proBNP levels increased significantly in patients with MS, but were significantly lower in those who underwent MVR. PMID- 16359053 TI - Prognostic significance of ischemic mitral regurgitation after non-Q-wave acute myocardial infarction. AB - BACKGROUND AND AIM OF THE STUDY: The development of mitral regurgitation (MR) soon after acute myocardial infarction (AMI) is a recognized and frequent complication. Its negative impact on survival has been observed after Q-wave AMI, even when of a mild degree, and independently of left ventricular systolic function. Few data exist regarding MR after non-Q-wave AMI (nQ AMI), however. Hence, the study aim was to investigate the incidence, clinical predictors and prognostic implications of MR in the setting of nQ AMI. METHODS AND RESULTS: A total of 99 consecutive patients (37 men, 62 women; mean age 72 +/- 13 years) who suffered a nQ AMI was studied. All patients underwent echocardiography during the first week after the nQ AMI. MR was detected in 34 patients (17 men, 17 women; mean age 76 +/- 10 years). Events during follow up were coded as death, AMI, unstable angina, or heart failure. The in-hospital outcome was not significantly different between patients with and without MR. The mean follow up period was 663 +/- 574 days. In the univariate analysis, freedom from hospital survival was significantly greater in patients without MR. However, multivariate analysis showed that MR was not an independent predictor of cardiovascular hospitalization or death. CONCLUSION: The incidence of MR is high among patients with nQ AMI but, unlike results found with Q-wave AMI, its presence does not add any prognostic significance to other known negative factors in the setting of nQ AMI. PMID- 16359054 TI - Hemolytic anemia produced by regurgitation through transposed chordae tendineae. AB - Hemolytic anemia after mitral repair and annuloplasty ring placement is very uncommon, and rarely described. The case is presented of a 53-year-old woman who developed severe mitral regurgitation and transfusion-dependent hemolytic anemia following mitral valve repair with a Carpentier-Edwards annuloplasty ring, which included transposition of chordae tendineae from the posterior leaflet to the anterior leaflet. Transesophageal echocardiography suggested that the transposed chordae tethered the anterior leaflet, causing malcoaptation of the leaflets. This resulted in central regurgitation divided by the chordae tendineae, producing two turbulent flow jets causing hemolysis. At reoperation, these chordae were removed and two longer Gortex neochordae to the anterior leaflet were placed with subsequent resolution of the anemia. To the authors' knowledge, this is the first case of hemolytic anemia caused by transposed mitral valve chordae tendineae from the posterior to the anterior leaflet. PMID- 16359055 TI - Results of valve preservation and repair for bicuspid aortic valve insufficiency. AB - BACKGROUND AND AIM OF THE STUDY: There is increasing interest in aortic valve sparing and repair techniques for the treatment of aortic insufficiency (AI) and/or root aneurysm. The results of bicuspid aortic valve (BAV) repair at the authors' institution were evaluated. An attempt was made to assess the mode of failure and to identify surgical methods that provide durable repair results. METHODS: Aortic valve repair for BAV was performed in 71 patients (62 men, nine women; mean age 41.5 +/- 13.2 years) between 1993 and 2005. Repair techniques included cusp free margin plication (n = 47), subcommissural annuloplasty (n = 27), free margin reinforcement (n = 10), and cusp triangular resection (n = 7). Thirteen patients had remodeling of one or both aortic sinuses, and 16 had reimplantation of the aortic valve. The ascending aorta and arch were replaced in 29 and five patients, respectively. Concomitant mitral and coronary bypass surgeries were performed in 10 and three patients, respectively. RESULTS: There were no operative deaths. Early postoperative echocardiography revealed no or trace AI in 54 patients and mild AI in 17. There was only one late death which was non-cardiac-related. Eight-year freedom from endocarditis, thromboembolism and anticoagulation-related hemorrhage was 90%, 100% and 100%, respectively. Eight-year freedom from AI grade > or =3+ (moderate) and aortic valve replacement were 44% and 82%, respectively. At the latest follow up, 89% of patients were in NYHA functional class I. Patients who underwent aortic valve-sparing procedures had more stable valve function than those who had cusp repair and subcommissural plication. CONCLUSION: BAV repair is a safe procedure with good early functional results. However, recurrent AI remains a problem at five to eight years of follow up. Since dilation of the aortic root is a common cause of AI and a common feature of patients with BAV, aortic valve-sparing reimplantation operations should provide better long-term outcomes. PMID- 16359056 TI - Outcome after valve replacement for severe aortic stenosis associated with reduced left ventricular ejection fraction. AB - BACKGROUND AND AIM OF THE STUDY: Because valve replacement for aortic stenosis (AS) remains a difficult surgical challenge in the presence of left ventricular dysfunction, the immediate and long-term outcomes, and evolution of left ventricular ejection fraction (LVEF) in this setting, were analyzed. METHODS: Forty-three consecutive patients with severe AS (valve area < or =1 cm2) and reduced LVEF (< or =40%) who underwent valve replacement surgery at the authors' institution between April 1998 and December 2003 and were studied retrospectively. RESULTS: Preoperative characteristics included: LVEF 33 +/- 6%, mean transaortic pressure gradient 46 +/- 13 mmHg, and aortic valve area 0.58 +/- 0.15 cm2. Concomitant coronary artery bypass grafting was performed in 15 patients (35%). Perioperative (30-day) mortality was 2.3%, with 39.5% morbidity. During a mean follow up of 33.4 +/- 17.6 months, eight patients died. The Kaplan Meier estimate of five-year survival was 75.3%. Postoperatively, none of the survivors remained in NYHA functional classes III-IV. The postoperative LVEF assessed in 81.8% of survivors had improved. Multivariate analysis associated improved LVEF with a higher preoperative mean transaortic pressure gradient (p = 0.0009) and a higher preoperative LVEF (p = 0.02). CONCLUSION: Patients with severe AS and reduced LVEF can undergo valve replacement with low perioperative mortality and moderate postoperative morbidity. Good long-term survival with good NYHA functional status and improved LVEF can be obtained. PMID- 16359057 TI - Reoperations for aortic aneurysm after the Ross procedure. AB - BACKGROUND AND AIM OF THE STUDY: Among late complications after the Ross operation, autograft dilatation is likely the most common. In order to define prevalence, consequences and management of autograft dilatation, a 10-year clinical experience was reviewed. METHODS: A total of 112 patients (mean age 29 +/- 10 years) underwent cross-sectional echocardiographic follow up. End-points of the study were freedom from autograft dilatation (diameter >4 cm, indexed as 0.21 cm/m2) and from reoperation for dilatation. Risk factors for autograft dilatation were also identified. RESULTS: There were 110 late survivors; average follow up was 5.1 +/- 1.9 years (range: 0.3 to 10.6 years). At 10 years, autograft dilatation was identified in 32 patients (29%), compatible with aortic aneurysm (>5.0 cm) in seven patients (6%). Seven of 32 patients (22%) presented moderate or greater autograft insufficiency. Ten-year freedom from dilatation was 43 +/- 8%, and from regurgitation was 75 +/- 8%. At multivariate analysis, preoperative aneurysm (p = 0.02), root replacement technique (p = 0.03) and absence of root buttressing (p = 0.04) were predictive of dilatation. Reoperation for autograft aneurysm was performed in five patients at a mean of 7.3 +/- 0.8 years after the Ross procedure, while two patients await reintervention. Two patients had root replacement and three remodeling with valve preservation (two root replacements, one sinotubular junction replacement): all survived reoperation. Ten-year freedom from root reoperation was 81 +/- 6%, and from full root replacement was 94 +/- 2%. CONCLUSION: With increasing follow up after the Ross operation, the incidences of root dilatation and reoperation are likely to rise. Graft replacement of coexisting aneurysm, avoidance of root replacement technique and the use of root-stabilization measures may reduce the prevalence of late root pathology. Early replacement of dilated autograft roots may allow preservation of the autologous pulmonary valve. PMID- 16359058 TI - Risk factors for congestive heart failure after aortic valve replacement with a Carpentier-Edwards pericardial prosthesis in the elderly. AB - BACKGROUND AND AIM OF THE STUDY: Congestive heart failure (CHF) after aortic valve replacement (AVR) is an important cause of morbidity. The study aim was to identify preoperative risk factors for CHF. METHODS: A total of 500 consecutive patients (271 males, 229 females; median age 73 years; range: 71-77 years) was investigated retrospectively. The AVR was performed using a Carpentier-Edwards pericardial valve, and a total of 348 additional procedures (313 coronary artery bypass grafts; CABG) was carried out. The outcome studied was CHF, during both hospital stay and long-term follow up. Univariate and multivariate statistical analyses were used to investigate 15 risk factors. RESULTS: During the hospital stay, 13 patients developed CHF, with four fatalities. Significant risk factors for CHF included urgent operation (p = 0.031), preoperative atrial fibrillation (AF) (p = 0.031) and NYHA functional class IV (p = 0.05). A logistic regression analysis revealed need for urgent operation (p = 0.034) as the sole factor. During long-term follow up, 43 patients developed CHF, with seven fatalities. Univariate analysis identified seven risk factors with significant effect: valve size <19 mm (p = 0.004), preoperative conduction defects (p = 0.007), chronic postoperative AF (p = 0.013), cross-clamp time >75 min (p = 0.032), NYHA class IV (p = 0.041), coronary artery disease (CAD) (p = 0.043) and additional CABG (p = 0.050). Multivariate analysis identified three risk factors: preoperative conduction defects (p = 0.004), postoperative AF (p = 0.005) and CAD (p = 0.037) CONCLUSION: Morbidity due to CHF after AVR could be minimized with correct treatment of AF and of conduction defects. Patient age, valve size, cross-clamp time and preoperative severity or symptoms were not independent risk factors. Moreover, small native aortic valve rings should not necessarily be enlarged, the cross-clamp time should be kept to a minimum, and surgery should not be delayed when symptoms have developed. PMID- 16359059 TI - An inverted left atrial appendage mimicking an intraatrial thrombus after a ross operation. AB - Inverted left atrial appendage (ILAA) is a rare manifestation. The case is reported of a patient with a history of factor V Leiden who underwent a Ross operation. An intra-atrial mass was discovered one month postoperatively and suspected to be a thrombus. Despite six months' anticoagulant treatment, the intra-atrial mass persisted. Ultimately, the patient was reoperated on due to hemostatic risks factors, and an ILAA was found. The diagnosis of ILAA remains a major challenge. Despite widespread use of postoperative echocardiography, the few reported cases confirm ILAA to be a rare phenomenon. The finding at post cardiotomy echocardiography of a left atrial mass associated with an absence of the left appendage is highly suggestive of a diagnosis of ILAA. As the incidence of complications remains to be defined, treatment remains controversial, but embolic risk factors such as hemostatic disease or atrial fibrillation invariably lead to surgery. In an asymptomatic patient without thromboembolic risks factors, a conservative approach should be considered, because of the risk of reoperation. PMID- 16359060 TI - Immunotherapy for refractory pulmonary infection after adult cardiac surgery: immune dysregulation syndrome. AB - BACKGROUND AND AIM OF THE STUDY: Pulmonary dysfunction/multiorgan failure (PD/MF), usually due to refractory pulmonary infection, is an important cause of mortality and morbidity after cardiac operations. Moreover, the incidence of PD/MF may be increasing due to the emergence of antibiotic-resistant pathogens. METHODS: Fifteen consecutive patients (median age 69 years) who were developing antibiotic-refractory PD/MF were administered 24 g per day intravenous immunoglobulin (IV-IgG; Carimune) for five days. Ten patients had undergone complex valve surgery, and five coronary bypass. Preoperatively, 93% of patients had significant comorbidity, 73% presented acutely, 53% were hypoalbuminemic and 47% had antecedent acute pulmonary derangement. Clinical variables were assessed by retrospective chart review for three days prior to (-3) the start of IV-IgG (day 0) and for five days afterwards (+5). A postoperative morbidity index (PMI) was generated as a weighted sum of: worsening lung infiltrates (I); leukocytosis (L); pulmonary dysfunction (P); ventilator requirement (V); septic shock (S); renal (R), gastrointestinal (G), or hepatic (H) dysfunction; thrombocytopenia (T); and delirium (D). RESULTS: At day 0, all patients were refractory to major antibiotics, with morbidities of: 1-100%, L-93%, P-93%, V-60%, S-27%, R-67%, G 40%, H-13%, T-27%, and D-20%. Using regression analysis, IV-IgG administration was associated with a statistically significant fall in white blood count and improvement in PMI (p <0.006). Fourteen patients (93%) recovered uneventfully, and one patient (7%) died from progressive sepsis. No complications of IV-IgG therapy occurred. CONCLUSION: Given the high predicted mortality of PD/MF patients, these data suggest that IV-IgG is a safe and efficacious adjunct to antibiotics in this setting. Further studies, including a randomized trial and investigation of immunomodulatory mechanisms, seem indicated. PMID- 16359061 TI - Early and late results of the surgical correction of cardiac prosthetic paravalvular leaks. AB - BACKGROUND AND AIM OF THE STUDY: Although the incidence of paravalular leaks, the most common cause of non-structural dysfunction after valve replacement, is well defined, the results of their surgical correction are not. Given the growing enthusiasm for interventional catheter-based correction of paravalvular leaks, a current surgical baseline against which to compare these results is important. METHODS: All patients who had surgical correction of an aortic or mitral paravalvular leak unrelated to acute bacterial endocarditis between 1986 and 2001 were identified from a computerized registry. Hospital records were reviewed, and follow up data obtained. RESULTS: A total of 136 consecutive patients (73 males, 63 females; mean age 64 years) underwent surgical correction of a paravalvular leak. Of the valves, 44 (32%) were aortic and 92 (68%) mitral. More than one previous cardiac operation had been performed in 68 patients (50%). In 107 patients (79%; 32 aortic (73%), 75 mitral (82%)), the leak was the primary indication for reoperation, while for 29 patients (21%; 12 aortic (27%), 17 mitral (18%)) the correction was secondary to another cardiac procedure. In 65 patients (48%; 12 aortic (27%), 53 (58%) mitral)) the leak was repaired primarily, while in 71 patients (52%; 32 aortic (73%), 39 (42%) mitral)) the prosthesis was replaced. Operative mortality was 6.6% (n = 9). There were no significant multivariable predictors of hospital death. Perioperative stroke occurred in seven cases (5.1%), and hospital stay was >14 days in 40 patients (29%). The 10-year Kaplan-Meier survival was 30 (CI 20-39)%. Ten-year actual versus actuarial freedom from repeat paravalvular leak was 84 (CI 68-92)% versus 63 (CI 49-76)%. CONCLUSION: Surgical correction of cardiac paravalvular leaks can be performed with acceptable mortality and morbidity. Patients with surgically corrected paravalvular leaks have a limited life expectancy, but reasonable freedom from recurrent paravalvular leak. PMID- 16359062 TI - Clinical features and outcome of non-drug-addicted patients with infective endocarditis and perivalvular abscess. AB - BACKGROUND AND AIM OF THE STUDY: Perivalvular abscess is a serious complication in infective endocarditis (IE) that confers a poor prognosis. Few data are available concerning the long-term outcome of these patients. The study aim was to determine clinical features and long-term prognosis of non-drug addict patients with IE complicated by perivalvular abscess. METHODS: Among a consecutive series of 241 patients with IE, 30 (12.4%) were diagnosed with perivalvular abscess and treated at the authors' institution over a 15-year period. A comparative analysis of patients with perivalvular abscess and other patients in the series was performed. RESULTS: Perivalvular abscess was more frequently associated with aortic valve endocarditis (93% versus 35%, p <0.05), and Streptococcus sp. was the predominant microorganism. Severe complications during hospital admission were more common in patients with perivalvular abscess (100% versus 61%, p <0.01). In-hospital mortality was significantly higher in patients with perivalvular abscess (33% versus 15%, p <0.05). Event-free survival at five years among survivors of the in-hospital phase was 86% in patients with perivalvular abscess, and 83% in those without abscess (p = NS). CONCLUSION: Patients with IE and perivalvular abscess have a higher in-hospital mortality rate, as major complications are more common in these patients. However, among patients who survived the active phase of the disease, long-term survival was similar with or without perivalvular abscess. PMID- 16359063 TI - Tissue engineering of cardiac valves: re-seeding of acellular porcine aortic valve matrices with human mesenchymal progenitor cells. AB - BACKGROUND AND AIM OF THE STUDY: Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study investigated the potential for re-seeding an acellular porcine heart valve matrix using human mesenchymal progenitor cells (MPC). METHODS: MPC were isolated from the bone marrow of patients undergoing hip replacement operations. Putative MPC were then cultured in several differentiation media in order to determine the multipotential differentiation capacity of the cells. The MPC were also characterized by FACS analysis. Cells at passage 8 were then seeded at between 1 x 10(4) and 1 x 10(5) cells/cm2 onto a decellularized porcine aortic valve matrix, and recellularization of the matrix was assessed. The phenotype of the re seeded cells and re-seeded cell density was then determined by histology and immunohistochemistry. RESULTS: Putative MPC were successfully isolated and differentiated into cells of the adipogenic, neurogenic, and myogenic lineages. FACS analysis showed the cells to have a similar phenotype to those isolated by others (CD45-, CD13+, D7FIB+, CD105+, CD10+/-, LNGFR+/-, CD55+, BMP- and AP+/-). Cells seeded onto an acellular valve matrix penetrated the center of the tissue after four weeks to 2% of homograft cell density. Phenotypic analysis of the cells in the re-seeded matrix revealed the cells to have a similar phenotype to native valve interstitial cells (vimentin+, alpha-smooth muscle actin+, heavy chain myosin slow-, desmin-). However, re-seeded cells also expressed osteogenic markers (alkaline phosphatase, osteonectin, and osteopontin). CONCLUSION: This study has shown, for the first time, that human MPC have the capacity to infiltrate an acellular porcine valve matrix under static conditions in vitro. Future studies will comprise culture under pulsatile flow in a physiological heart valve bioreactor to maintain the desired cell phenotype and increase cell density. PMID- 16359064 TI - The Toronto root stentless valve in the subcoronary position is hemodynamically superior to the mosaic stented completely supra-annular bioprosthesis. AB - BACKGROUND AND AIM OF THE STUDY: Stentless valves are considered to exhibit better hemodynamics after aortic valve replacement (AVR) compared to stented valves. However, a new generation of stented bioprostheses for completely supra annular implantation has been designed to optimize the ratio of the effective orifice area (EOA) of the prosthesis and aortic annulus area. The study aim was to determine whether a stentless valve implanted in the subcoronary technique renders larger orifice areas and lower transvalvular pressure gradients at rest and exercise compared to a completely supra-annular stented device. METHODS: Twenty patients underwent AVR for aortic stenosis with the St. Jude Medical (SJM) Toronto Root stentless porcine bioprosthesis, using a subcoronary implantation technique. Through the authors' institutional database, 20 additional patients were identified who had undergone AVR with the Medtronic Mosaic stented completely supra-annular porcine bioprosthesis. The patient groups were not matched for labeled valve size, but for annulus diameter measured intraoperatively using Hegar's dilators. Hemodynamic performance was assessed by transthoracic echocardiography at discharge (early) and by rest and stress echocardiography at six months postoperatively (mid-term). RESULTS: Transvalvular mean pressure gradients (MPG) at rest were significantly lower in the stentless group, but cardiac output was similar in both groups. Stress echocardiography also revealed significantly lower gradients at 25 W and 50 W exercise in the stentless group. The EOA index (EOAI), grouped by annulus diameter, tended to be larger in the stentless group and showed no severe patient-prosthesis mismatch (PPM; EOAI <0.65 cm2/m2) which, in contrast, occurred in three patients (15%) in the stented group (p = 0.072). CONCLUSION: In summary, the SJM Toronto Root porcine stentless bioprosthesis in the subcoronary position showed lower MPGs and larger EOAs at rest and during exercise compared to the Medtronic Mosaic porcine stented bioprosthesis. Therefore, physically active patients in particular may benefit from use of the stentless valve. Because of its larger EOA, a stentless valve should be implanted if severe PPM is expected. PMID- 16359065 TI - Hemodynamic evaluation of the Sorin Soprano bioprosthesis in the completely supra annular aortic position. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate the clinical and hemodynamic performance of the Sorin Soprano bioprosthesis in the aortic position. METHODS: Rest and stress echocardiography were performed at six months after surgery in 57 patients who underwent aortic valve replacement with the stented Soprano bioprosthesis. The exercise protocol included workloads of 25, 50, 75 and 100 W, each of 2 min duration. RESULTS: Thirty-day mortality was 1.8% (n = 1). Due to malperfusion of the coronary arteries, two Soprano prostheses had to be replaced by a different prosthesis, and one patient received coronary artery bypass grafts. One patient developed bacterial endocarditis of his prosthesis at seven months postoperatively and died as a result of a fulminant sepsis. There were no other prosthesis-related adverse events. Mean pressure gradients (MPG) ranged from 7.8 to 15.9 mmHg, effective orifice areas (EOA) from 1.25 to 2.98 cm2, EOA index (EOAI) from 0.79 to 1.43 cm2/m2, and EOA fraction from 34 to 45%. Stress echocardiography showed no significant increase in MPG up to 50 W, and MPGs did not exceed 35 mmHg at 75 and 100 W. CONCLUSION: Initial hemodynamic results showed low MPGs during rest and exercise. The EOAI was large due to completely supra-annular placement of the Soprano valve. The difficulty of complete supraannular placement is that the prosthesis is positioned relatively high, especially in narrow aortic roots, and this may handicap coronary artery perfusion. Patient selection with suitable aortic root anatomy is crucial to achieve the benefit of completely supra-annular implantation without coronary ostium deterioration. PMID- 16359066 TI - Hemodynamic performance of the Ultracor and Carpentier-Edwards aortic prostheses using exercise and dobutamine stress echocardiography. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to compare the hemodynamic profiles of the aortic standard porcine Carpentier-Edwards (C-E) and Ultracor (tilting-disc) valve using exercise and dobutamine stress. METHODS: A total of 36 patients was examined, 18 for each valve type. When analyzing the data, valve types were matched for valve size, which ranged from 21 to 25 mm. All patients were analyzed within an 18-month period after implantation. Hemodynamic variables, two-dimensional echocardiography and Doppler flows were recorded at rest. These were repeated during bicycle ergometry, performed for a maximum of four, 3-min stages, with each stage increasing in workload by 20 W. After resting, patients were subjected to dobutamine stress, administered up to a maximum 40 microg/kg/min, dependent upon heart rate. RESULTS: Mean (+/- SD) resting pressure drops across the C-E and Ultracor valves were comparable (maximum drop 19.4 +/- 8.6 versus 22.9 +/- 12.2 mmHg; mean drop 9.96 +/- 3.8 versus 11.83 +/- 6.6 mmHg, respectively). During exercise, the maximum cardiac flow rate attained was approximately 400 ml/s for both valve types. At this flow rate, the maximum and mean pressure differences between valve types were 6.2 mmHg and 4.4 mmHg, respectively (p = NS). During dobutamine stress, the maximum cardiac flow attained was approximately 500 ml/s, which resulted in significant differences between valve types of 11.6 and 7.3 mmHg, for maximum and mean pressure drops, respectively. When slopes of the mean pressure drop/cardiac flow were calculated for individual valves, a difference was observed between the two valve types (p = 0.02 and p = 0.039 for dobutamine and exercise, respectively). CONCLUSION: Both prostheses demonstrated significant increases in pressure drop under stress conditions. The standard porcine C-E valve had a statistically better hemodynamic profile than the Ultracor prosthesis at higher flow rates. When a study cohort of patients is small, these differences will only be evident at optimal flow rates. The higher flow rates seem most easily obtained when using pharmacological stress. PMID- 16359067 TI - Surgical excision of papillary fibroelastoma for known or potential embolization. AB - BACKGROUND AND AIM OF THE STUDY: Papillary fibroelastoma (PFE) is a rare and histologically benign tumor, but it may have malignant propensity for life threatening complications. Herein are described four cases of PFE which reflect the clinical diversity of this lesion. The diagnostic and surgical approach utilized is also briefly reviewed. METHODS: The files of four patients with cardiac valvular PFE treated at the authors' center between January 2002 and November 2003 were reviewed. The diagnosis was strongly suggested by echocardiography. Tumors were noted in aortic (n = 2), mitral (n = 1) and tricuspid (n = 1) sites. Indications for surgery were myocardial infarction (both aortic tumors), previous stroke (mitral tumor), and preventive (tricuspid tumor). RESULTS: Surgical excision with a conservative, valve-sparing approach was performed in all cases. For the first aortic tumor, the aortic valve was reconstructed with a patch of autologous pericardium. None of the patients had evidence of valvular regurgitation after excision on intraoperative transesophageal echocardiography, and all had an uneventful recovery. There were no cases of recurrence or regurgitation on follow up echocardiography. CONCLUSION: PFE is an uncommon but increasingly recognized cause of embolic phenomena. Prompt identification allows for surgical excision, which seems to be curative, safe and well-tolerated. A conservative valve-sparing approach is recommended because of the absence of recurrence after total excision. PMID- 16359069 TI - Traumatic tricuspid regurgitation resulting in protein-losing enteropathy: a case report. AB - Protein-losing enteropathy (PLE) with hypoproteinemia is an uncommon but serious complication of congestive heart failure. Reports of patients with PLE resulting from severe tricuspid regurgitation (TR) caused by trauma are rare. A 66-year-old male diabetic patient had a chest contusion as a result of a road traffic accident, and one year later suffered from progressively generalized edema. Examination revealed severe TR with a high central venous pressure and PLE with serum protein deficiency. Treatment with albumin administration and diuretic therapy proved ineffective, and consequently the TR was corrected by valve replacement. Postoperatively, the serum protein level gradually returned to normal. Surgical intervention successfully improved this patient with severe TR and PLE resistant to medical treatment. PMID- 16359068 TI - Infected multiple fibroelastomas in hypertrophic cardiomyopathy. AB - Cardiac papillary fibroelastomas are rare benign tumors of the heart with potential for life-threatening complications. The incidence of multiple lesions is less than 10% of all reported cases. Preoperative transesophageal echocardiography is important for detecting all cardiac sites involving this tumor, because excision of all such tumors must be performed to prevent serious complications. Here, the first ever case is reported of multiple infected papillary fibroelastoma of the mitral valve, aortic valve and left ventricular outflow tract with massive mitral insufficiency in a patient with hypertrophic cardiomyopathy. PMID- 16359070 TI - Ross-Konno procedure and ostial plasty in a child with homozygous hypercholesterolemia: case report. AB - Although the Ross procedure is preferred for aortic valve replacement in young and female patients, there are no reported cases of hypercholesterolemic aortic pathology due to homozygous familial hypercholesterolemia. Long-term durability of the pulmonary autograft in patients with postoperative high blood cholesterol levels is of interest. A 14-year-old girl with homozygous familial hypercholesterolemia who underwent the Ross-Konno procedure and left coronary artery ostial plasty was followed for 57 months, with pulmonary autograft function, coronary arteries and lipid profile being monitored. There were no signs of narrowing, insufficiency or calcification of the pulmonary autograft; neither was there any narrowing in the left main coronary ostium. The patient's total cholesterol level was reduced from 897 to 262 mg/dl by use of anti lipidemic medication and weekly lipid apheresis. Follow up data suggest that a pulmonary autograft may be preferable in children with hypercholesterolemic aortic valvular pathology, as well as in children with aortic valvular diseases of other etiologies. PMID- 16359071 TI - Factors associated with non-bacterial thrombotic endocarditis: case report and literature review. AB - The case is reported of non-bacterial thrombotic endocarditis (NBTE) in a patient with giant cell arteritis and prostate cancer, and the relevant literature reviewed. To the present authors' knowledge, this is the first case to be reported where NBTE coexists with both arteritis and cancer. NBTE is difficult to diagnose if the underlying disease is accompanied by fever. If a diagnosis of probable infective endocarditis is established, the cultures and serology are negative, and there is no response to antibiotic treatment, then NBTE should be considered. PMID- 16359072 TI - Riding the gene train. PMID- 16359073 TI - The patient voice in infertility. PMID- 16359074 TI - Premature births hit record high. PMID- 16359075 TI - Hereditary Breast and ovarian cancer: providing personalized medicine. PMID- 16359076 TI - Collaborative initiatives for postpartum depression. PMID- 16359077 TI - Tummy time! Preventing unwanted effects of the "Back to Sleep" campaign. PMID- 16359078 TI - Pregnancy after bariatric surgery. PMID- 16359079 TI - The perplexing parity puzzle. PMID- 16359080 TI - Fetal health surveillance: the use of research evidence in practice. PMID- 16359081 TI - Recruitment and Retention strategies: creating the best workplace for women's services. PMID- 16359082 TI - Drug interactions: beyond polypharmacy and P-450... PMID- 16359083 TI - AWHONN nurses respond to Katrina. PMID- 16359084 TI - What parents of near-term infants need to know. PMID- 16359085 TI - Two views of the same picture. PMID- 16359086 TI - Mom was a bad cook. PMID- 16359087 TI - The evolution of ethics in Indiana: back to the future. PMID- 16359088 TI - Dentistry and Indiana law. PMID- 16359089 TI - TRUST: the foundation of a profession. PMID- 16359090 TI - How to react ethically when an unethical situation arises. PMID- 16359091 TI - Ethical dilemmas: they don't call them sticky situations for nothing. PMID- 16359093 TI - Meth mouth: a national scourge. PMID- 16359094 TI - Responding to Katrina. PMID- 16359095 TI - Nursing school: new challenges, new rewards. PMID- 16359096 TI - Study: who are the men in nursing? AB - Many of the closing comments that respondents shared revisited areas of concern and optimism about the nursing profession and men's place within it. Emphasis was placed on the need to recruit more men and minorities into the profession, confront stereotypes, and improve the general awareness and knowledge of what the profession has to offer to all its members. Just as in the student survey, we were struck by how articulate and passionate respondents were about nursing. Countless respondents indicated that nursing was a 'calling'; that they especially enjoyed 'making a difference' and how unique nursing is for that reason. We also appreciated the humor and self-confidenc they demonstrated about the reality of being a minority in the profession, and their 'just do it' mindset. PMID- 16359097 TI - A fish out of water. PMID- 16359098 TI - The importance of mentoring. PMID- 16359099 TI - Nurse educators can empower future nurse leaders through role modeling. PMID- 16359100 TI - My journey. PMID- 16359101 TI - Establishing a student mentoring program. PMID- 16359102 TI - The impact of learning in a mentorship community. PMID- 16359103 TI - How to recruit minority students into nursing. PMID- 16359104 TI - Behavioural and physiological responses of three chicken breeds to pre-slaughter shackling and acute heat stress. AB - 1. The aim of this study was to compare the behavioural and physiological responses to hanging and acute heat stress in three different chicken breeds. Chicks were obtained from a slow-growing French 'Label Rouge' line (SGL), a fast growing standard line (FGL) and a heavy line (HL). The SGL, FGL and HL birds were slaughtered at their respective market ages of 12, 6 and 6 weeks, in an attempt to achieve similar body weights. Before stunning, birds were either shackled by their legs on the moving line for 2 min (shackling stress: SH) or placed in a room at 35 degrees C and 60% of humidity for 3.5 h and then shackled for 2 min (acute heat stress plus shackling: H + SH) or subjected to minimal stress by shackling for 10 s before stunning (control group: C). 2. Bird physiological responses to the three pre-slaughter treatments were estimated by measuring blood corticosterone, glycaemia, creatine kinase activity, acid-base status and electrolyte concentration as well as lactate content and glycolytic potential in the breast (Pectoralis major) and thigh (Ilio tibialis) muscles. Behavioural responses to shackling stress were evaluated by measuring wing flapping duration, straightening up attempts and vocalisations. 3. Blood corticosterone was higher in SH and H+SH groups than in the C group, regardless of genotype. The struggling activity on the shackle line differed among chicken breeds. It was more intense and occurred more rapidly after hanging in the SGL birds than in both other breeds. Furthermore, SGL struggling activity was not affected by hanging duration while it increased with hanging duration in FGL and HL birds. 4. Wing flapping duration was negatively correlated with blood pH, bicarbonate concentration and positively correlated with breast muscle lactate content, indicating that struggling stimulated antemortem glycolysis activity in breast muscle. Acute heat stress affected blood Ca2+ and Na+ concentration and increased glycaemia and glycolytic potential of thigh muscle. 5. Both acute heat stress and shackling before slaughter were experienced as stressful events by all types of birds. PMID- 16359105 TI - Keeping laying hens in furnished cages and an aviary housing system enhances their bone stability. AB - 1. Tibia and humerus breaking strength of Lohmann Silver hybrids kept in conventional cages, furnished cages and an aviary with outdoor run were examined in two production cycles. Each trial lasted a full laying period; feeding, management and healthcare were identical for all hens. In both trials bone strength was investigated at the end of laying months 6, 9 and 14. 2. The objective was to determine if bone strength increases when hens are kept in alternative housing systems, especially in furnished cages, and whether hen age affects bone stability. 3. The results indicated that housing system influenced bone breaking strength, which was consistently higher for hens in the aviary compared to hens in conventional and furnished cages. Furthermore, humerus breaking strength was higher for hens in furnished cages compared to conventional cages. No significant difference regarding tibia breaking strength was found between conventional and furnished cages. 4. Our results showed that lack of exercise contributed to the problem of weak bones more than did calcium depletion from eggshell formation. 5. Tibia breaking strength increased during the last third of the production cycle, whereas humerus breaking strength remained unaffected by hen age. 6. Genetic group affected only tibial bone breaking strength, which was lower overall in genetic group A than in group B, which in turn was lower than group C. 7. The increased bone strength in the aviary and in the furnished cages probably reduced the incidence of recently broken bones in these systems compared to the conventional cages. This increase in bone strength can be regarded as an improvement in welfare. Furnished cages, like the aviary system, might be considered an alternative housing system for laying hens, because both resulted in enhanced bone strength. PMID- 16359106 TI - Assessing the welfare consequences of providing litter for feed-restricted broiler breeders. AB - 1. Broiler breeder females were fed restricted allocations of a standard wheat soy ration to meet target body weights. They were housed on raised plastic slotted floors (S) or wood shavings litter (L) from hatch to 8 weeks when each pen of 12 birds was transferred to another pen in the same block in a 2 x 2 factorial experiment (LL, LS, SL and SS). Measures of bird welfare were taken at 4 weeks of age, and at 9 and 10 weeks following transfer to the new pens. 2. At 4 weeks of age, birds reared on S spent more time standing, pecking the feeder, the wall and other birds and less time pecking the floor compared with those on L. Gentle feather pecks, strong feather pecks and strong feather pulls were more common in S than L. 3. Mean body weight was higher and coefficient of variation lower in birds on L than S at the end of the experiment. Feather loss and damage scores at 10 weeks were higher for birds reared from hatch to 8 weeks on S. 4. Birds that were reared on S continued to peck more at the pen walls after transfer to new pens but there was no other carry-over effect on behaviour. Birds on S at 9 and 10 weeks pecked more at the walls and less often at the floor, and rested less often. There was more feather pecking on S than on L in the second week post transfer. 5. Tonic immobility was greater and plasma corticosterone concentrations were lower at the end of the experiment in birds on L than S at 9 and 10 weeks of age. The heterophil-lymphocyte ratio was similar between treatments at 4 weeks and after the birds were moved to a new pen. 6. The results are consistent with the view that litter and wall pecking has de-arousing properties and that this activity is re-directed foraging that diminishes the stress of feed restriction. PMID- 16359107 TI - Effect of furnished small group housing systems and furnished cages on mortality and causes of death in two layer strains. AB - 1. Mortality and causes of death were recorded over two trials with 13045 Lohmann Selected Leghorn (LSL, white layer strain) and 4410 Lohmann Brown (LB) laying hens kept in furnished small group housing systems and furnished cages. In trial 1, hens were housed from weeks 19 to 71 (LSL) and 17 to 69 (LB). In trial 2, LSL hens were housed from weeks 18 to 57. 2. The hens were kept in groups of 40 and 60 in the furnished small group housing system Eurovent 625 + a-EU and in groups of 10 and 20 in the furnished cages Aviplus and Eurovent 625A-EU. 3. To investigate the possible effect on mortality of rearing the birds on litter floors or in cages, in trial 2 half of the LSL hens were reared on the floor and half in conventional cages. 4. Mortality was greater in the Eurovent furnished small group housing system (5.2%) than in the furnished cages Aviplus (4.0%) and Eurovent (4.2%). 5. Mortality was also affected by the interactions among housing system and trial, layer strain and rearing. In trial 1, more LB laying hens died in the Aviplus system than in the Eurovent group system. There was no significant difference in mortality of the LSL laying hens between housing systems in trial 1. In trial 2, mortality of the LSL hens was greater in the group housing system (hens reared on litter floors: 10.1%; hens reared in cages: 2.8%) than in the furnished cage Aviplus (hens reared on litter floors: 6.0%; hens reared in cages: 1.5%). 6. Mortality was greater in floor-reared LSL laying hens than in the LSL laying hens reared in conventional cages. 7. Group size within housing system had no significant effect on mortality. 8. The most common findings at autopsy were different types of cannibalism (65.51%), with vent cannibalism (38.57%) the most common. PMID- 16359108 TI - Yolk sac, body dimensions and hatchling quality of ducklings, chicks and poults. AB - 1. One-day-old chicks of Pekin duck, turkey, layer fowl and broiler fowl were examined for bacteria in the yolk sac and yolk fluid. 2. Whole hatchling, yolk free hatchling and yolk sac weights were recorded for all species along with crown-rump length and beak-tip to toe-tip length. 3. Bacteriology revealed positive results for the whole yolk sacs of 43 to 64% of the birds in the sample of ducklings, poults and layer chicks. Broiler chicks had a 6.6% incidence of bacteria isolated from the whole yolk sac. By contrast, there were very few positive results from swabs of yolk fluid for any of the bird types. 4. The presence of bacteria in the yolk sac of hatchlings suggests that there is colonisation, rather than infection, of the yolk sac membrane during the hatching period or the first few hours post-hatching. Isolation of bacteria from the yolk sacs of young chicks might no longer be considered as solely indicative of yolk sac infection but further research is required to confirm this result. 5. Contrary to what is being suggested in commercial practice relationships between linear dimensions and hatchling weight suggest that measurement of chick length is at best a very crude measure of chick quality. PMID- 16359109 TI - Selection responses in duration of fertility and its consequences on hatchability in the intergeneric crossbreeding of ducks. AB - 1. From 1992 to 2003, selected (S) and control lines (C) of the laying Brown Tsaiya duck (Anas platyrhynchos) were simultaneously maintained under the same standardised conditions of feeding and management. 2. The selection objective was to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. From generations G1 to G11, 2452 and 2022 female ducks, in S and C lines, respectively, were measured and recorded. In the S line, the percentage selected varied between 20.2 and 34.3% in females and between 7.2 and 20.8% in males. 3. Selection for number of fertile eggs had a correlated effect of increasing the parameter tau of the logistic curves which fitted the daily variations (d 2 to 15) in fertility or hatchability on the basis of eggs set. The differences S-C for the estimates of the times of half maximal fertility and hatchability increased by 0.41 and 0.37 d per generation between G1 and G11, respectively. 4. The highest increases of fertility per day rates after a single AI were observed between d 5 and 11. Moreover, in the selected line, fertility rate was higher than, or equal to, 90% in d 2 from G8. The same tendencies were observed for the changes in the evolution of hatchability on the basis of eggs set. 5. Selection increased fertility and hatchability according to the egg set rates, especially for d 2 to 8 after AI. Hatchability of fertile eggs was not impaired, confirming that selection for one AI per week was possible in this strain of laying ducks. PMID- 16359110 TI - Variations in chicken breast meat quality: implications of struggle and muscle glycogen content at death. AB - 1. Pectoralis major (P. major) muscle pH and meat quality traits were studied in relation to bird response to ante-mortem stress in three chicken lines: a fast growing standard line (FGL), a slow-growing French 'Label Rouge' line (SGL) and a heavy line (HL). Ninety-nine birds of the three genetic types were slaughtered at their usual marketing age (6, 12 and 6 weeks for FGL, SGL and HL birds, respectively) on the same day. The birds of each line were divided into three different ante-mortem treatment groups: minimum stress (shackling for 10 s) (C), shackling for 2 min (SH) and acute heat plus shackling stress (exposure to 35 degrees C for 3.5 h and shackling for 2 min before stunning) (H + SH). 2. Regardless of chicken line, wing flapping duration (WFD) between hanging and stunning was strongly negatively related to P. major muscle pH at 15 min post mortem. It was also moderately negatively related to P. major muscle glycolytic potential (GP), which represents glycogen level at death. Increasing WFD induced an increased ultimate pH (pHu) only in HL. The consequences of increased WFD for breast meat traits were dependent on the chicken line: it induced lower L* and b* and higher a* and drip loss in SGL while it only increased breast a* in HL birds. By contrast, WFD variations did not alter breast meat quality traits of FGL birds. Regardless of the chicken line, increased GP was associated with lower pHu and higher L* and drip loss. In SGL, it also increased b* and decreased curing cooking yield of breast meat. 3. Struggling activity on the shackle line and muscle glycogen content at death could partly explain line and pre-slaughter variations in breast meat pH and quality traits. The water holding capacity of the raw and cooked meat was impaired by long shackling in the case of SGL birds while it was barely affected by ante-mortem conditions in the two standard lines. In conditions which minimised bird struggling (C), SGL and FGL birds had meat with a better water holding ability than that of broilers from the heavy line. However, when broilers were subjected to SH or H + SH conditions, the breast meat water holding capacity of SGL birds was lowered to the same level as that of the heavy line birds. PMID- 16359111 TI - Residues of gentamicin in eggs following medication of laying hens. AB - 1. Gentamicin was injected subcutaneously and intramuscularly into 5 groups of 10 laying hens and its concentration was determined in albumen, yolk and whole egg. 2. Groups 1 and 3 were intramuscularly injected with doses of 10 and 25 mg/kg while groups 2, 4 and 5 were subcutaneously injected with doses of 10, 25 and 50 mg/kg, respectively. 3. The final gentamicin concentration in albumen was measured on d 3 for groups 1 and 2; on d 4 for groups 3 and 4, and on d 5 for group 5. Concentrations in yolk and whole egg were measured on d 7, 10 and 12. 4. Gentamicin recovery was as follows: 2% in groups 1 and 2, 2.5% in groups 3 and 4, and 3% in group 5. 5. Most of the residue (approximately 90%) was recovered from the yolk. PMID- 16359112 TI - Change in the effect of constant photoperiods on the rate of sexual maturation in modern genotypes of domestic pullet. AB - 1. Data for modern egg-type hybrids reared on constant daylengths show that, as expected, they mature more quickly than earlier genotypes. However, the constant photoperiod which gives earliest sexual maturity has not changed as a result of selection and is 10 h for both early and modern genotypes. 2. Further analysis showed that the rate of delay in sexual maturity for constant photoperiods above 10 h is similar for modern and for early hybrids (+0.29 d for each incremental one hour of photoperiod), the response of modern hybrids below 10 h (+4.22 d for each one-hour reduction in photoperiod) is more than double that of early hybrids (+1.71 d/h). PMID- 16359113 TI - Effect of amount and source of supplemental dietary vegetable oil on broiler chickens exposed to aflatoxicosis. AB - 1. Addition of sunflower oil (SFO) at 30 or 60 g/kg or three vegetable oils, namely SFO, soybean (SBO) or groundnut (GNO), at 30 g/kg to isocaloric and isonitrogenous broiler chicken diets were evaluated for possible counteractive effects against aflatoxin (AF) (0.3 microg B1/g diet) from 0 to 42 d of age. 2. Body weight, food intake and serum concentration of protein were lower in the AF group than in the control, whereas in the SFO and SBO supplemented groups they were comparable with those of the control. Sunflower oil at both concentrations exerted similar effects on growth. Groundnut oil did not improve growth or food intake in AF-fed birds. 3. The serum concentration of cholesterol and triglycerides decreased with AF feeding and was increased by supplementation of any of the three oils both in the control and in AF-fed groups. 4. Liver and giblet weight and liver fat content were increased by AF; these effects were countered by dietary oil inclusion, except for liver weight at 60 g/kg SFO. Weights of pancreas and gall bladder were increased by AF. Oil supplementation reduced the weight of pancreas in chickens given AF. 5. Humoral immune response was depressed by AF and dietary oil supplementation (particularly SFO or SBO) countered this effect. Other variables, namely, serum gamma glutamyl transferase activity, bone mineralisation, weights of lymphoid organs, kidney and adrenals, ready-to-cook yields and fat content in muscle and skin showed little or no effect of dietary oil supplementation. 6. It is concluded that dietary inclusion of SFO or SBO at 30 g/kg may alleviate the adverse effects of 0.3 microg/g of AF B1 in commercial broiler chickens. Groundnut oil, although showing beneficial effects on some biochemical variables, failed to improve growth performance. PMID- 16359114 TI - Effect of dietary dried oregano leaves on growth performance, carcase characteristics and serum cholesterol of female early maturing turkeys. AB - 1. A study was conducted with 120 female early maturing turkeys to test the effect of dietary dried oregano leaves (Origanum vulgare subsp. hirtum) on body weight (BW), feed intake (FI), feed conversion efficiency (FCE), carcase characteristics and serum cholesterol concentration. Dried oregano leaves had a content of 3.6 ml essential oils/100 g, while the carvacrol content was 855 g/kg of the total essential oils. 2. From 1 to 84 d of age, the turkeys were fed on 4 diets varying in oregano content (OR0, no oregano--control; OR45, 1.25 g oregano/kg; OR90, 2.5 g oregano/kg; OR135, 3.75 g oregano/kg). Birds were given feed and water ad libitum. 3. BW was unaffected by oregano throughout the experiment. FI and FCE were similar among all treatments until 42 d of age. From 43 to 84 d of age and for the overall experimental period, FI decreased linearly in treatment OR135 and FCE increased linearly with dietary oregano content. Body and carcase weights, carcase yield, and the relative weights of the heart and liver were not significantly affected by oregano content. The relative weights of the gizzard and small intestine decreased linearly with oregano content. Serum cholesterol content was similar among all treatments. 4. In the present study, dietary oregano (1.25, 2.5 and 3.75 g/kg) improved FCE in female early maturing turkeys between 43 and 84 d, with the lowest oregano inclusion (1.25 g/kg) giving the most cost effective diet. Thus, dried oregano leaves may be used as a natural herbal growth promoter for early maturing turkeys. PMID- 16359115 TI - A mixture of pure cellulase, hemicellulase and pectinase improves broiler performance. AB - 1. The effects of a mixture of pure enzymes (cellulase, hemicellulase and pectinase) and a commercial enzyme, Energex, were examined on performance and metabolisabilities in broiler chicks given a maize-soybean meal diet. Composition of the mixed enzyme was similar to Energex except that protease was not present. 2. Chicks were divided into three groups: control, mixed enzyme and Energex with 7 replicates per group. Male broiler chicks were raised at 25 degrees C in wire floored cages for 12 d from 15 d of age. Feed and water were offered ad libitum. 3. The Energex group gained significantly more weight and the mixed enzyme group tended to gain more than the control. Feed intakes were similar and thus the feed conversion ratio of Energex was significantly improved while it tended to be improved by the mixed enzyme. 4. The mixed enzyme group showed significant improvement in carcase and muscle weight when compared with the control group. The mixed enzyme group also showed significant improvement in organic matter and crude protein metabolisabilities. In the groups given enzyme, abdominal fat weight tended to decrease. 5. It is concluded that a combination of cellulase, hemicellulase and pectinase is effective in improving organic matter and crude protein metabolisabilities and carcase yield of broilers on a maize-soybean meal diet. PMID- 16359116 TI - Metabolisable energy and amino acid digestibility of high-oil maize, low-phytate maize and low-phytate soybean meal for White Pekin ducks. AB - 1. Two experiments were conducted to determine the metabolisable energy and amino acid digestibilities of high-oil maize (HOM), low-phytate maize (LPM) and low phytate soybean meal (LPSBM) as compared with normal maize (NM) and normal soybean meal (NSBM) using male White Pekin ducks. 2. In the first experiment, the TMEN (kJ/g) value of HOM (16.58) was higher than that of NM (16.05), but that of LPM (16.11) did not differ from those of HOM or NM. The true digestibility coefficients for methionine (0.874, 0.871 or 0.876), lysine (0.805, 0.816 or 0.813) and tryptophan (0.946, 0.959 or 0.960) did not differ among NM, LPM and HOM, respectively. The average true digestibility coefficients for all amino acids in NM, LPM or HOM (0.886, 0.890 or 0.900, respectively) did not differ among the three ingredients. 3. In the second experiment, the TMEN value of LPSBM (12.39) was approximately 8% higher than that of NSBM (11.46). The true digestibility coefficients for most amino acids were higher for LPSBM than for NSBM, except in the case of histidine, cysteine and tyrosine, which were not different. The average true digestibility coefficient for all amino acids in LPSBM (0.945) was higher than in NSBM (0.924). 4. Results of the two experiments indicate that the energy and amino acids in the feed ingredients evaluated were well utilised by ducks and that LPSBM has a higher energy value as well as digestible essential amino acid concentration than NSBM for ducks. PMID- 16359117 TI - Pharmacokinetics and tissue residues of pefloxacin and its metabolite norfloxacin in broiler chickens. AB - 1. The pharmacokinetics of pefloxacin and its active metabolite norfloxacin were investigated in chickens after a single oral administration of pefloxacin at a dosage of 10 mg/kg. To characterise the residue pattern, another group of chickens was given 10 mg of pefloxacin/kg body once daily for 4 d by oral route; the tissue concentrations of pefloxacin and norfloxacin were determined at 1, 5 and 10 d after the last administration of the drug. 2. The concentrations of pefloxacin and norfloxacin in plasma and tissues were determined by HPLC assay. The limit of detection for pefloxacin and norfloxacin was 0.03 microg/ml in plasma or microg/g in tissue. 3. The plasma concentration-time data for pefloxacin and norfloxacin were characteristic of a one-compartment open model. The elimination half-life, maximum plasma drug concentration, time to reach maximum plasma drug concentration and mean residence time of pefloxacin were 8.74 +/- 1.48 h, 3.78 +/- 0.23 microg/ml, 3.33 +/- 0.21 h and 14.32 +/- 1.94 h, respectively, whereas the respective values of these variables for norfloxacin were 5.66 +/- 0.81 h, 0.80 +/- 0.07 microg/ml, 3.67 +/- 0.21 h and 14.44 +/- 0.97 h. 4. Pefloxacin was metabolised to norfloxacin to the extent of 22%. 5. The concentrations of pefloxacin (microg/g) 24 h after the fourth dose of the drug declined in the following order: liver (3.20 +/- 0.40) > muscle (1.42 +/- 0.18) > kidney (0.69 +/- 0.04) > skin and fat (0.06 +/- 0.02). Norfloxacin was also detectable in all the tissues analysed except muscle. No drug and/or its metabolite was detectable in tissues except skin and fat 5 d after the last administration. The concentrations of pefloxacin and norfloxacin in skin and fat 10 d after the last dose of pefloxacin were 0.04 +/- 0.02 and 0.03 +/- 0.01 microg/g, respectively. PMID- 16359118 TI - Can testosterone and corticosterone predict the rate of display of male sexual behaviour, development of secondary sexual characters and fertility potential in primary broiler breeders? AB - 1. Genetic selection for growth to enhance production may be associated with stress and with modified physiological and behavioural phenotypes which depress male primary broiler breeder fertility. 2. We hypothesised that male serum testosterone (T) and corticosterone (C) concentrations might correlate with fertility, sexual behaviour, and testicular, comb and wattle size. 3. Cockerels from two genetic strains (A and B) of primary broiler breeder were penned individually with an average of 10 females across 5 age periods (30 to 51 weeks) to evaluate male fertility, behaviour, serum T and C, and comb, wattle and testicular dimensions. 4. Strain A males had higher T at age periods 2, 4 and 5 than Strain B. Both strains had basal concentrations of C, apart from an elevated concentration for Strain B in period 5. 5. Strain B had a weak but significant, positive correlation between sexual behaviour and T and C, while Strain A males with higher C had larger combs and wattles. 6. Neither T nor C correlated with fertility. We conclude that evaluation of these endocrine factors (quantifiable measurements with the potential to correlate with fertility) alone seems insufficient to predict male fertility potential in these strains of primary broiler breeder. PMID- 16359119 TI - Effects of dietary coenzyme Q10 supplementation on hepatic mitochondrial function and the activities of respiratory chain-related enzymes in ascitic broiler chickens. AB - 1. One hundred and sixty 1-d-old Arbor Acre male broiler chicks were fed with maize-soybean based diets for 6 weeks in a 2 x 2 factorial experiment. The factors were CoQ10 supplementation (0 or 40 mg/kg) and Escherichia coli lipopolysaccharide (LPS) challenge (LPS or saline). 2. CoQ10 was supplemented from d 1. From d 18, the chickens received three weekly i.p. injections of LPS (1.0 mg/kg BW) or an equivalent amount of sterile saline as control. From d 10 on, all chickens were exposed to low ambient temperature (12 to 15 degrees C) to induce ascites. 3. The blood packed cell volume and ascites heart index of broiler chickens were reduced by dietary CoQ10 supplementation. Mitochondrial State 3 and State 4 respiration, respiratory control ratio and phosphate oxygen ratio were not changed, but H+/site stoichiometry of complex II + III was elevated by dietary CoQ10 supplementation. 4. Cytochrome c oxidase and H+-ATPase activity were increased by CoQ10 supplementation, whereas NADH cytochrome c reductase and succinate cytochrome c reductase were not influenced. Mitochondrial anti-ROS capability was increased and malondialdehyde content was decreased by CoQ10 supplementation. 5. The work suggested that dietary CoQ10 supplementation could reduce broiler chickens' susceptibility to ascites, which might be the result of improving hepatic mitochondrial function, some respiratory chain related enzymes activities and mitochondrial antioxidative capability. PMID- 16359120 TI - Single administration of xylitol to newly hatched chicks enhances growth, digestive enzyme activity and immune responses by 12 d of age. AB - 1. An experiment was conducted to determine the effect of a single administration of xylitol to newly hatched chicks on growth, digestive enzyme activity and immune responses at 12 d of age. 2. Female broiler chicks (Cobb) were given 0.5 ml of either 20% glucose, 20% xylitol or water alone within 24 h after hatch. Thereafter, all chicks were reared under conventional conditions and given a commercial broiler starter diet until 12 d of age in experiment 1. In experiment 2, they were deprived of feed and water for 24 h and kept at 27 degrees C to mimic transportation stress before given feed and water. 3. Body weights at 5 d of age did not differ among the treatments, but at 12 d of age chicks given 20% xylitol showed greater body weight than those in the other treatment groups. Bursa weight (mg per 100 g body weight) was greater in chicks given xylitol solution than in chicks given glucose solution or water at 5 and 12 d of age. 4. Amylase and chymotrypsin activities in the pancreas of chicks given xylitol solution were higher than in chicks given water or glucose solution at 5 and/or 12 d of age. 5. A single administration of xylitol within 24 h after hatch increased splenocyte proliferation against concanavalin A and pokeweed mitogen, and antibody titres to keyhole limpet haemocyanin (KLH) and Mycobacterium butyricum (Mb) as compared to glucose, administration at 12 d of age, but not as compared to water administration. 6. These results indicated that xylitol may be a functional carbohydrate source to improve growth rate and health and to relieve transportation stress in broiler chicks. PMID- 16359121 TI - Clinicopathology of gout in growing layers induced by high calcium and high protein diets. AB - 1. An experiment was conducted to test the independent and combined effects of high dietary calcium and protein concentrations on the induction of visceral gout in growing birds of a layer strain. 2. One hundred and sixty healthy birds were randomly divided into 4 groups at 35 d of age. The different groups were given 4 diets containing normal or high concentrations of dietary calcium or crude protein in a 2 x 2 factorial experiment for 30 d. The diets contained normal calcium (Ca) and crude protein (CP) (NCNP, 8.5 g Ca/kg and 175g CP/kg), high calcium and normal protein (HC, 36.3 g Ca/kg and 175 g CP/kg), normal calcium and high protein (HP, 8.8 g Ca/kg and 245 g CP/kg) or high calcium and high protein (HCHP, 36.8 g Ca/kg and 242 g CP/kg), respectively. 3. Typical visceral gout was induced by the HCHP diet. The HCHP and HC diet caused severe kidney damage. The HP diet did not cause kidney damage, but significantly increased plasma uric acid and inorganic phosphorus concentrations. 4. The HC diet significantly increased plasma uric acid, calcium and sodium, but significantly decreased plasma inorganic phosphorus, potassium and magnesium concentrations. The HCHP diet significantly increased plasma uric acid, calcium and sodium. 5. Urine volumes were significantly higher on the HCHP and HC diets than on the control. The growers raised on HC and HCHP diets had significantly higher total quantity of 24 h urinary excretion of uric acid, calcium, magnesium, inorganic phosphorus and potassium and a significantly lower 24 h urinary excretion of sodium. The growers fed on the HP diet had a higher 24 h urinary excretion of uric acid and inorganic phosphorus than the control. 6. It is concluded that growing layer birds should not be fed on layer rations. PMID- 16359122 TI - Selenium, a versatile trace element: current research implications. AB - Selenium (Se), a trace element, has evolved from its toxic properties to an essential element. Se was known a potent antioxidant through glutathione peroxidase (selenium being part of this molecule). Later, many other selenium binding proteins were discovered and their functions were tried to be known with unsuccessful results in many cases. Se is known to be involved in carcinogenesis, immune function, male reproduction, cardiovascular diseases etc. The specific mechanism of the involvement of the element is still not known. Recent research with application of modern research tools viz. bioinformatics, cDNA microarray and transgenesis have revealed the mechanism of selenium involvement in various processes. This review highlights mysterious and useful roles of selenium in biological processes. PMID- 16359123 TI - Gangliosides enhance migration of mouse B16-melanoma cells through artificial basement membrane alone or in presence of laminin or fibronectin. AB - The migration of B16LuF1 cells, B16-melanoma cells of lower metastatic potential to lung was enhanced through artificial basement membrane in presence of gangliosides of B16LuF1 cells as well as gangliosides of B16-melanoma cells of higher metastatic potential to lung, namely, B16LuF5 and B16LuF10 cells. The same concentration (50 microM) of gangliosides of B16LuF1, B16LuF5 and B16LuF10 cells gradually increased the migration of B16LuF1 cells through basement membrane. Moreover, B16LuF10 cell gangliosides modified the migratory effect of laminin and fibronectin on B16LuF1 cells. Laminin alone increased migration of B16LuF1 cells whereas fibronectin alone decreased migration of the same cells. When B16LuF10 cell gangliosides were used in combination with fibronectin, gangliosides removed the migration inhibitory effect of fibronectin resulting in net enhancing effect. Gangliosides in association with laminin also increased the enhancing effect of laminin on migration of B16LuF1 cells. Thus, gangliosides showed additive enhancing effect when used in combination with laminin. However, effect of individual gangliosides were different. Out of six gangliosides isolated from B16LuF10 cells only two gangliosides corresponding to standard gangliosides GM2 and GM3 enhanced migration of B16LuF1 cells. The migration of B16LuF1 cells in presence of each of the remaining four gangliosides corresponding to GT1b, GD1b, GD1a and GM1 was not altered and was comparable to that of untreated control. Thus, gangliosides of B16 melanoma cells alone or in combination with laminin or fibronectin enhanced migration of B16 melanoma cells through artificial basement membrane, suggesting possible role of tumor gangliosides during invasion of metastatic tumor cells through basement membrane of the surrounding tissues in vivo. PMID- 16359124 TI - Modulatory effects of different doses of alpha-tocopherol on benzo(a)pyrene-DNA adduct formation in the pulmonary tissue of cigarette smoke inhaling mice. AB - Cigarette smoke (CS) has been established as one of the major risk factors for many pathologies including lung cancer in humans and experimental animals. In view of the discrepancy about the role of alpha-tocopherol (AT) in carcinogenesis, the present study was designed to investigate the effects of different doses of AT on benzo(a)pyrene-DNA [B(a)P-DNA] adduct formation in lungs of CS inhaling mice. Extent of carcinogen-DNA adduct formation has been considered as an index for carcinogenesis. Feeding of 35 IU AT/kg body weight increased B(a)P-DNA adducts formation significantly whereas feeding of 5 IU AT/kg body weight did not altered much the B(a)P-DNA adduct levels when both were compared to the control counterparts. With CS inhalation, the B(a)P-DNA adducts formation increased in all the groups when compared to their respective sham counterparts. Interestingly, in CS exposed groups, there was least increase in B(a)P-DNA adducts formation in 5 IU AT/kg fed animals followed by the control and 35 IU AT/kg body weight fed groups respectively. The results suggest that higher doses of AT accentuate DNA adduct formation in CS inhaling mice. PMID- 16359125 TI - Kinetics of humoral immune response in pigs vaccinated against foot and mouth disease. AB - The present investigation was conducted to study the foot and mouth disease virus (FMDV)-specific humoral immune response (HIR) in pigs, following vaccination with oil adjuvanted foot and mouth disease (FMD) vaccine, upto 90 days post vaccination (dpv). For this, 40 Large White Yorkshire (LWY) pigs (20; one-year old female (gilts) and 20; three-month old piglets) were vaccinated @ 2 ml/animal, subcutaneously. Sera samples were collected at fortnight interval from all the animals. The log10 SN50 antibody titres against all the serotypes (Type O, A and Asia-1) were detected in both gilts and piglets from day 7 to 90 dpv indicating the persistence of HIR up to the last day of sampling. The maximum antibody titres were observed on 28 dpv, thereafter, titres started declining, but were present till 90 dpv against all the three FMDV serotypes. HIR was more pronounced in piglets in comparison to gilts, as group mean SN antibody titres against all the three FMDV serotypes were found to be more maintained and significantly higher in piglets. PMID- 16359126 TI - Effect of 5-lipoxygenase inhibitor against lipopolysaccharide-induced hypothermia in mice. AB - Bacterial endotoxin produces sepsis associated with alterations in body temperature (fever or hypothermia). The intraperitoneal administration of bacterial endotoxin, lipopolysaccharide (LPS; 50 microg/mouse) led to a decrease in colonic temperature starting 1 hr after the injection. The hypothermic effect was accompanied by a significant increase in hypothalamic leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) levels. 5-lipoxygenase inhibitor, zileuton (200 and 400 mg/kg, po) administered 30 min before LPS challenge significantly prevented hypothermia. However, non-selective cyclooxygenase inhibitor, indomethacin (10, 20 mg/kg, po) did not reverse the hypothermic response. Further, pretreatment of mice with zileuton prevented LPS-stimulated increase in hypothalamic LTB4 levels and caused a relatively small increase in PGE2 levels. Indomethacin had no effect on LTB4 levels but it reduced PGE2 levels. These results suggest a possible involvement of leukotrienes in LPS-induced hypothermia and the potential protective role of 5-lipoxygenase inhibitors in endotoxemia. PMID- 16359127 TI - Antiviral profile of Nyctanthes arbortristis L. against encephalitis causing viruses. AB - The ethanolic extracts, various fractions and two pure compounds isolated from the plant N. arbortris were tested against Encephalomyocarditis Virus (EMCV) and Semliki Forest Virus (SFV). Pronounced in vitro virus inhibitory activity was observed with the ethanolic and n-butanol fractions as well as with the pure compounds arbortristoside A and arbortristoside C. In addition, ethanolic extracts and n-butanol fraction protected EMCV infected mice to the extent of 40 and 60% respectively against SFV at a daily dose of 125 mg/kg body weight. PMID- 16359128 TI - Beneficial effects of Zingiber officinale Roscoe on fructose induced hyperlipidemia and hyperinsulinemia in rats. AB - Fructose supplementation produced cardinal features of Syndrome-X including significant elevations in seum cholesterol, triglyceride, glucose and insulin and also in body weight. While treatment with methanolic extract of dried rhizomes of Zingiber officinale produced a significant reduction in fructose induced elevation in lipid levels, bodyweight, hyperglycemia and hyperinsulinemia, treatment with ethyl acetate extract of Z officinale did not poduce any significant change in either of the last two parameters. However, it produced a significant reduction in elevated lipid levels and body weight The concentration of 6-gingerol was found to be higher in methanolic extract and less in ethyl acetate extract. The results suggest that the methanolic extract of Z officinale produces better effects as compared to ethyl acetate extract in fructose induced hyperlipidemia associated with insulin resistance. The extent of activity appears to be dependent on the concentration of 6-gingerol present in the extracts. PMID- 16359129 TI - Protective role of Spirulina feed in a freshwater fish (Poecilia reticulata Peters) exposed to an azo dye-methyl red. AB - Acute toxicity of an azo dye-methyl red (5-40 ppm) was examined under starving conditions, on two groups of Poecilia reticulata--a freshwater fish, fed on different diets prior to their exposure to dye. Besides natural feed, fish of group-1 also received Spirulina feed for one month (feed population), whereas those of group-2 received only natural feed (non-feed population). The mortality data revealed non-feed population to be more tolerant to feed stress during acute toxicity study, whereas feed population exhibited better tolerance to the combined stress of both feed and methyl red; especially at higher concentrations of the latter. RBCs in methyl red treatments acquired different shapes (poikilocytosis) and an increase in their size (anisocytosis) was also noticed. Percentage of such abnormal RBCs was almost equal in both feed and non-feed populations, except at a lower concentration (5 ppm), at which percentage of poikilocytic RBCs was lesser in the feed population. RBC counts in the control non-feed fish (34.5 x 10(4)/mm3) were significantly lower than control feed population (50.0 x 10(4) /mm3). Their number decreased with an increase in methyl red concentrations in non-feed population (9-26%), but percent reduction in RBC counts was almost similar (20-26%) at various concentrations of methyl red (5-30 ppm) in the feed population. Despite reduction in RBC counts, feed population did not suffer from anemia in methyl red treatments, as evident by their RBC counts which were almost equal to control fish of non-feed population. The results suggest that Spirulina feed improves tolerance of test organism towards methyl red manifested by noticeable reduction in the cytotoxic effects on RBCs and a lower mortality rate at higher concentrations of dye. PMID- 16359130 TI - Identification of allergens in Indian fishes: hilsa and pomfret exemplified by ELISA and immunoblotting. AB - Enzymed-linked immunosorbent assay of hilsa and pomfret muscle extracts showed specific IgE binding to ten allergic patients' sera, the results corroborated to that of skin prick test. Comparison of allergen profiles of the two fish extracts by immunoblotting revealed a common antigenic protein of 50 kDa and some high molecular weight fish allergens instead of low molecular weight parvalbumin found in several fishes. Purified and well characterized fish allergens are always considered better than crude fish extracts for diagnostic use. PMID- 16359131 TI - Modulatory influence of juvenile hormone analogue (JHa) and 20-hydroxyecdysone on lipophorin synthesis in red cotton bug, Dysdercus cingulatus Fabr. AB - Topical supply of methoprene, a juvenile hormone analogue (JHa) caused notable morphological disturbance in insects. Topical supply of methoprene to newly emerged adult female D. cingulatus caused notable disturbance and induced a dramatic reduction in the total haemolymph protein pattern and lipophorin production in tissues like fat body, ovary and haemolymph. Total protein concentration in haemolymph also showed significant reduction in 1 day old insects but increased slightly as age advanced. Application of 20-hydroxyecdysone (20-HE) to 2-day-old adult female stimulated protein synthesis intensively. Lipophorin levels in fat body and ovary also simultaneously increased. Densitometric analysis revealed that methoprene inhibits while 20-HE stimulates lipophorin production in D. cingulatus. PMID- 16359132 TI - Mode of utilization of amino acids as growth substrates by Azospirillum brasilense. AB - The study was undertaken to analyze the rate of uptake and utilization of various amino acids by Azospirillum brasilense Sp81 (RG) in a basal mineral salts solution under non-nitrogen fixing condition. These amino acids including other nitrogenous compounds were tested for both N- and C-sources. The kinetic constants (Km and Vmax) of uptake of some amino acids (e.g. lysine, arginine, proline, glutamine and glutamic acid) were exploited using a Hanes-Woolf plot, and discussed in the context of nitrogen starvation or both carbon and nitrogen starvation. To summarize all the kinetic data for these amino acids strongly suggested that the mode of these amino acids utilization in this bacterium followed the same general pattern, although the quantitative differences were there. A single amino acid was able to satisfy the nitrogen needs of this bacterium in basal mineral salts solution, and this possibility could be considered for the cost-effective growth medium for this bacterium in the biotechnological industry. PMID- 16359133 TI - NaCl induced changes in photosystem stoichiometry and photosynthetic activity of the cyanobacterium Microcystis aeruginosa Kutz. AB - Exposure to 0.4 M NaCl resulted in higher PS I/PS II stoichiometry and increase in the rate of photosynthesis in planktonic cyanobacterium M. aeruginosa. Altered ratios of PS I/PS II as well as photosynthesis and respiration were stabilized within 72 hr of exposure to salt, leading to adaptation of the organism to the changed conditions. PMID- 16359134 TI - Effectivity of crude versus purified mycobacterial secretory proteins as immunogen for optimum antibody production. AB - Monospecific antibodies have been successfully utilized in antigen detection, which is better indicator of active infection. Mycobacterium tuberculosis excretory secretory (M tb ES) antigens such as ES 31, ES 41 and ES 43 (31 kDa, 41 kDa and 43 kDa protein, respectively) have been shown to be present in Mycobacterium tuberculosis H37Ra culture filtrate and are of diagnostic interest. To study the immunogenic potential of crude versus purified antigen, goat was immunized with M tb detergent soluble sonicate (DSS) antigen as well as purified antigen fraction (ESAS 7) containing ES 31 antigen. Both anti-DSS IgG antibody and anti ESAS 7 IgG antibody were found to be reactive with ES 31 antigen upto 1 ng concentration of antibody by ELISA. Crude DSS antigen was found to be quite effective in producing high titre antibodies and showed further high reactivity with other ES antigens (ES 41 and ES 43) of diagnostic interest. PMID- 16359135 TI - Bilateral hearing loss following electrocution. AB - Electrocution is a rare cause of sensorineural deafness. We describe and discuss the first reported case in this country of domestic electrocution resulting in a bilateral sensorineural deafness. PMID- 16359136 TI - Pulsatile tinnitus. AB - Pulsatile tinnitus is an uncommon otological symptom. Objective pulsatile tinnitus has numerous causes, including benign intracranial hypertension, glomus tumours and atherosclerotic carotid artery disease. History and physical examination can give important information as to the underlying diagnosis. Radiographic evaluation is essential in all patients with pulsatile tinnitus. Many patients have a treatable underlying aetiology. Early diagnosis and appropriate intervention may save patients from unnecessary morbidity. PMID- 16359137 TI - Outcomes in the use of intra-tympanic gentamicin in the treatment of Meniere's disease. AB - The pathogenesis of Meniere's disease is associated with a disorder of ionic homeostasis, with the pathologic correlate being endolymphatic hydrops. Despite uncertainty as to its particular mode of action, it is accepted wisdom that intra tympanic gentamicin has a definite therapeutic role in the control of symptoms in patients who fail to respond to medical therapy. This study reports an evaluation of the efficacy of intra-tympanic gentamicin in the treatment of Meniere's disease and also presents a simple, reliable, safe method of administering gentamicin for this purpose. A retrospective review of 56 patients undergoing intra-tympanic gentamicin treatment for Meniere's disease was conducted. Response to treatment was analysed using a patient survey and examination of pure-tone averages. An overall significant improvement in vertigo symptoms of 81.3 per cent was found. There was a 21.4 per cent rate of significant hearing loss, defined as greater than 10 dB, with an average loss in this group of 18.5 dB. A single dose of gentamicin applied directly to the round window resulted in a high rate of control of vertigo, with acceptably low rates of hearing loss. PMID- 16359138 TI - Follow-up results in tuberculous cervical lymphadenitis. AB - OBJECTIVE: To investigate the efficacy of medical antituberculous treatment in patients with tuberculous cervical lymphadenitis (TCL). METHODS: In the period 1996-2002, 73 TCL patients were reviewed and the results of clinical and laboratory testing were documented. The efficacy of a four-drug chemotherapy regimen was investigated. RESULTS: Purified protein derivatives (PPD) skin test results were positive in 58 (79 per cent) patients. Chest X-rays revealed changes consistent with tuberculosis in nine (12.3 per cent) patients. The mean duration of medical treatment was 10.04 months. In follow-up evaluation, 14 (20 per cent) patients were considered suspicious for resistant TCL and total excision of all nodes was performed. Histopathology confirmed TB in only 10 of these cases. CONCLUSION: The high incidence of residual disease in our study indicates that medical treatment (at least nine months of four combined antituberculous drugs) did not seem to be effective. If lymphadenopathy persists, total surgical excision of lymph nodes should be the treatment of choice. PMID- 16359139 TI - Small cell neuroendocrine carcinoma in the petrous apex. AB - With the exception of moderately differentiated neuroendocrine carcinoma of the larynx, neuroendocrine carcinomas are very rare head and neck malignancies. We report a case of a small cell neuroendocrine carcinoma in the petrous apex of the temporal bone. This unusual site for this tumour has not yet been reported in the literature. We confirmed the location and extent of the primary tumour by positron-emission tomography scan. The final histopathological diagnosis was of small cell carcinoma, and this was confirmed by immunohistochemistry. We treated this patient with surgery followed by radiotherapy and chemotherapy. After the treatment was completed there was subtotal remission of the tumour, with no distant metastases. PMID- 16359140 TI - Cochlear implantation in a patient with profound hearing loss with the A1555G mitochondrial DNA mutation and no history of aminoglycoside exposure. AB - The A1555G mitochondrial deoxyribonucleic acid (mtDNA) point mutation has classically been associated with sensorineural hearing loss in patients following aminoglycoside exposure. More recently, the mutation has been implicated in sensorineural hearing loss in patients without previous aminoglycoside use. In addition, cochlear implantation has been shown to be effective in the group of patients with prior aminoglycoside exposure but, to date, no case of cochlear implantation in a patient with the A1555G mutation and no prior exposure to aminoglycosides has been explicitly described in the literature. We report the case of an 80-year-old woman with the A1555G mtDNA mutation, a 35-year history of bilateral progressive hearing loss and no history of aminoglycoside exposure who underwent successful implantation of a Nucleus 24 Contour device at our institution. Post-operatively, the patient exhibited marked improvement in tests of auditory performance. We conclude that cochlear implantation can be an effective method to restore some sense of hearing in patients with the A1555G mtDNA mutation and sensorineural hearing loss. PMID- 16359141 TI - Successful treatment of MRSA otorrhoea: a case report. AB - Infection with methicillin-resistant Staphylococcus aureus (MRSA) is increasing. It may be community or hospital acquired and is characteristically difficult to eradicate. Here we report a case of a two-year-old girl who sustained a traumatic tympanic membrane perforation following a minor burns injury. She was seen as an out-patient in a burns unit and subsequently developed MRSA otorrhoea. This was treated with a two-week course of fusidic acid topical drops. At three-week follow up the tympanic membrane had healed and the infection had healed completely. Fusidic acid is safe and effective in the treatment of MRSA otorrhoea. We need to maintain vigilance in the treatment of otorrhoea, as MRSA may become an increasingly common pathogen in the future. PMID- 16359142 TI - Sarcomatoid salivary duct carcinoma of the larynx. AB - The sarcomatoid variant is a rare subtype of salivary duct carcinoma that comprises epithelial and sarcomatoid components. The authors present the first reported case of sarcomatoid salivary duct carcinoma occurring in the larynx, which was successfully treated by partial laryngectomy with ipsilateral neck dissection, with no evidence of recurrence after five years. Microscopic analysis revealed a composite epithelial and sarcomatoid tumour. The epithelial component was characterized by papillary proliferation of glandular and ductal structures lined with multilayered epithelial tumour cells. The sarcomatoid component was characterized by pleomorphic spindle cells with cytologic atypia, accompanied by small lymphoid cells, extensive proliferation of blood vessels and focal myxoid changes. To the best of our knowledge, only one case of sarcomatoid salivary duct carcinoma outside a major salivary gland has been reported previously in the literature, and this is the first report of sarcomatoid salivary duct carcinoma arising from the larynx. PMID- 16359143 TI - Swallowing dysfunction in patients with unilateral vocal fold paralysis: aetiology and outcomes. AB - Although unilateral vocal fold palsy (UVFP) is a common problem, data relating to swallowing dysfunction are sparse. We reviewed the clinical findings (method of presentation, underlying diagnosis and position of the vocal folds) of 30 patients and conducted a follow-up telephone survey. Outcome measures used were direct visualization of fold function, position and compensation. In addition, standardized speech and language assessments for swallowing dysfunction and dysphonia were noted and compared to presentation. Our study indicates that 56 per cent of patients with UVFP have associated dysphagia. Outcome with speech therapy is significant, with 73 per cent showing improvement. These data indicate a significant link between UVFP and swallowing dysfunction. There is a marked therapeutic benefit from voice therapy. Further work is required to evaluate the long-term outcomes and establish the mechanism of swallowing dysfunction in these patients. PMID- 16359144 TI - Day-case major ear surgery: is it viable? AB - INTRODUCTION: At present there is a strong emphasis on carrying out more elective day-case surgery. This paper presents a series of major ear cases performed on a day-case basis. METHOD: This is a retrospective review of outcome. Surgery was performed over 10 years at the Queen Elizabeth Queen Mother Hospital (part of the East Kent NHS Trust, UK). RESULTS: Over 10 years 253 major ear cases were performed. The unplanned admission rate for overnight stay was 2.8 per cent and the rate of short-period transfers to in-patient wards (for less than a six-hour stay) was 2 per cent. CONCLUSIONS: Our experience shows that major ear surgery, including mastoidectomy, is safe on a day-case basis in all age groups. With the advent of new anaesthetic agents recovery is faster, and modern anti-emetic cover for post-operative nausea and vomiting makes nearly all such major ear surgery feasible as day-case procedures. PMID- 16359146 TI - Neck dissection: past, present and future? AB - With the exception of distant metastasis, the presence of cervical lymph node metastasis is the single most adverse independent prognostic factor in head and neck squamous cell carcinoma. Surgical removal of metastatic cervical lymph nodes had been attempted during the late nineteenth century, with varying techniques and poor results. A systematic approach to en bloc removal of cervical lymph node disease, described in detail by Jawdynski at the end of the nineteenth century and popularized and illustrated by Crile in the early twentieth century, provided consistent and more effective treatment and forms the basis of our current techniques. The concepts of radical neck dissection, employed extensively by Martin, were followed with almost religious consistency by most head and neck surgeons until the late twentieth century, when the principles of 'functional' neck dissection, developed by Suarez and popularized by Bocca, Gavilan, Ballantyne, Byers and others, led to the acceptance of modified radical neck dissection as treatment for lymph node disease in various stages. More recently, selective neck dissection, involving removal of nodes confined to the levels at greatest risk of metastasis from primary tumours at various sites, has become accepted practice for elective and, in some instances, therapeutic treatment of the neck. In the future, sentinel lymph node biopsy and the use of molecular pathological analyses may be employed to predict the presence of occult cervical disease, thus directing therapy to patients at greatest risk and sparing those without regional metastasis. PMID- 16359145 TI - CO2 laser myringotomy in children with otitis media with effusion. AB - BACKGROUND: The study aimed to evaluate the clinical application of CO2 laser myringotomy in children with otitis media with effusion (OME) under topical anaesthesia in an office setting. METHODS: Laser myringotomy was performed with the CO2 laser Otoscan (OtoLAM) in 54 children (73 ears) with OME. The procedure on the tympanic membrane was performed under topical anaesthesia using Bonain's solution or 10 per cent Xylocaine (lidocaine) solution for 30 minutes before surgery. A circular perforation was created with a power of 15 W, single pulse duration of 200 msec and a scanned area of 1.9 mm in diameter. RESULTS: The mean healing time was 2.51 weeks (range 1-5 weeks). Effusion content was not a predictive prognostic factor for perforation healing time. Perforation location over anterior inferior or posterior inferior quadrants was not a predictive factor for perforation healing time. Xylocaine was the more effective anaesthestic. The OME resolution rate was 73 per cent. CONCLUSION: Laser myringotomy provides intermediate duration middle-ear ventilation. It could be beneficial in selected children with OME. PMID- 16359147 TI - Lipomas of the head and neck: presentation variability and diagnostic work-up. AB - INTRODUCTION: Head and neck lipomas have seldom drawn attention in the literature, except in isolated case reports. AIMS: This study aimed to assess the presentation variability of head and neck lipomas as well as the relative importance and efficiency of pre-operative diagnostic methods used. MATERIALS AND METHODS: A retrospective review was undertaken of medical records and imaging studies of 24 patients with histopathologically proven head and neck lipomas, over a three-year period. RESULTS: The 24 patients had 26 lipomas. Men predominated (62.5 per cent). The posterior subcutaneous neck was the most common site. Three patients had deep lipomas affecting the hypopharynx, larynx and parotid gland; all were correctly diagnosed pre-operatively. Computed tomography (CT) scan with specific radiodensity recording was the preferred pre-operative investigation. CONCLUSIONS: Lipomas should be considered in the differential diagnosis of soft-tissue head and neck masses even in rare locations. A CT or magnetic resonance imaging scan can correctly diagnose a lipoma pre-operatively, thereby allowing better treatment planning. PMID- 16359148 TI - Chondrodysplasia punctata: case report and review of audiological and ENT features. AB - Chondrodysplasia punctata is a term referring to a clinically heterogeneous group of bone and cartilage dysplasias which cause characteristic epiphyseal stippling. The condition can involve the ear, nose and throat in diverse ways at many levels. We present a case of X-linked brachytelephalangic chondrodysplasia punctata, which illustrates the features of this condition particularly relevant to the audiological physician, otolaryngologist and neonatologist. PMID- 16359149 TI - Transnasal flexible laryngo-oesophagoscopy: an evaluation of the patient's experience. AB - The aim of this study was to evaluate the patient's experience of transnasal flexible laryngo-oesophagoscopy under local anaesthetic in an out-patient setting. This was a clinical observational study using a patient questionnaire and visual analogue score. Subjects were 50 patients seen in the hospital's ENT outpatient clinic between March and August 2004 in whom transnasal flexible laryngo-oesophagoscopy was performed and who completed pain discomfort questionnaires. Any patient undergoing transnasal flexible laryngo oesophagoscopy, for either diagnostic or therapeutic purposes, was included in the study. Patients in whom transnasal flexible laryngo-oesophagoscopy was not necessary as part of their investigation or treatment were excluded. Patients completed a questionnaire, providing a simple visual analogue score for discomfort during the procedure. Any operative complications were noted by the surgeon. All patients completed transnasal flexible laryngo-oesophagoscopy, and the upper aerodigestive tract was clearly visualized. The procedure is well tolerated, with mean score of <1 out of 10 for all forms of discomfort. There was a complication rate of 2 per cent (one patient with epistaxis); no other complications occurred. Transnasal flexible laryngo-oesophagoscopy is a new diagnostic and therapeutic technique which is well tolerated by patients. It compares favourably with other flexible endoscopic techniques. This report documents for the first time a detailed description of patients' experience of this technique. PMID- 16359150 TI - Influence of anaesthetic agents on transient evoked otoacoustic emissions and stapedius reflex thresholds. AB - This aim of this study was to determine the effect of anaesthetic agents on stapedius reflex (SR) thresholds and transient evoked otoacoustic emissions (TEOAE). Fifty patients who were scheduled for operation and who had normal hearing were included in the study. All were given midazolam for premedication and propofol for induction. Anaesthesia was maintained in five different ways in each group of 10 patients. Groups I-IV received inhalational anaesthesia: group I received 70 per cent N2O plus 30 per cent O2, group II sevoflurane, group III desflurane and group IV halothane. Group V received total intravenous anaesthesia with propofol plus sufentanil. The SR and TEOAE of the patients were measured four times: on the day before surgery (first measurement), after premedication (second measurement), after induction of anaesthesia (third measurement) and during maintenance of anaesthesia (fourth measurement). Midazolam significantly increased ipsilateral and contralateral SR thresholds and decreased TEOAE wave reproducibility. Propofol significantly increased only the SR thresholds. The other anaesthetic agents significantly increased only the contralateral reflex thresholds. Of these, the highest increase was seen after sevoflurane and the lowest after halothane. The changes in TEOAE wave reproducibility due to anaesthetic agents used for maintenance were not significant. We concluded that midazolam premedication may affect audiological evaluation with SR and TEOAE tests, and sevoflurane should not be used when it is necessary to measure SR under general anaesthesia. PMID- 16359151 TI - Otoacoustic emissions in young adults with a history of otitis media. AB - The aim of this study was to investigate otoacoustic emissions in young adults who had a history of otitis media (OM) in childhood and to assess whether a history of OM had an irreversible effect on hearing. We studied 116 cases between 15 and 25 years of age, divided into three groups. Each subject underwent a single examination comprising otoscopy, pure-tone audiometry (PTA), tympanometry, and transient evoked otoacoustic emission (TEOAE) and distortion product otoacoustic emission (DPOAE) testing. Subjects in the first and second groups had normal audiometric hearing thresholds and type A tympanograms. The only difference between the first and second group was the presence or absence of a history of OM. The third group consisted of patients diagnosed as having active OM; these patients had poorer hearing thresholds and type B or C tympanograms. After statistical analysis of TEOAE and DPOAE results (one-way analysis of variance test), significant differences were noted between groups. Otoacoustic emission levels were, unsurprisingly, lowest in the third group, as expected. However, the most striking result in the study was that significantly fewer otoacoustic emissions were detected in subjects with a history of OM than in subjects without a history of OM. These findings suggest that OM in childhood may cause minor but irreversible damage to the middle ear or cochlea. Otoacoustic emissions testing can be used to detect this sub-clinical damage. PMID- 16359152 TI - Glioneural hamartoma of the VIIIth nerve. AB - Hamartomas of the cerebellopontine angle or internal auditory canal are very rare and only four cases have been reported. We report an unusual case of a glioneural hamartoma of the VIIIth nerve with clinical, radiological and audiometric similarity with vestibular schwannoma. PMID- 16359154 TI - Nitinol stents in the treatment of benign proximal tracheal stenosis or tracheomalacia. AB - Nitinol stents have been used in the treatment of benign tracheal stenosis. A retrospective review of five patients treated at Stobhill Hospital over the last six and a half years is presented. Age at presentation ranged from 17 to 76 years. The minimum follow-up period was 23 months and the maximum was 78 months. All our patients were successfully decannulated, with none requiring recannulation. Four patients developed granulation tissue related to the stent at intervals ranging from three weeks to 41 months post stenting. Topical mitomycin C application has been useful after resection of granulations using the carbon dioxide (CO2) laser. Stent migration occurred in one patient three weeks after insertion. Nitinol stents are easy to insert and effective in the treatment of tracheal stenosis, but can have associated morbidity. Their use should be considered carefully, as insertion should be regarded as permanent. Publications reporting experience and outcome with the use of Nitinol stents in the trachea are reviewed. PMID- 16359155 TI - Nasal histoplasmosis in the acquired immunodeficiency syndrome. AB - Disseminated histoplasmosis is a disease with a high case-fatality rate, especially in patients with the acquired immunodeficiency syndrome (AIDS). The disease can occur in various sites, such as the lungs, eyes, oral cavity, larynx, nervous system, gastrointestinal tract and, more rarely, the nasal sinus region. It is a cosmopolitan mycosis with a high prevalence in Brazil. Nasal manifestation of the disease is rare, with only three cases reported in the literature, but it is part of the differential diagnosis for other granulomatous diseases, such as Wegener's granulomatosis, tegumentary leishmaniasis and nasal lymphoma. The authors of this study present a literature review and report a case of nasal histoplasmosis in a patient with AIDS. No record of such an aggressive presentation has been reported previously in the literature. PMID- 16359156 TI - Rigid endoscopy in globus pharyngeus: how valuable is it? AB - The aim of this study was to assess the value of rigid endoscopy in patients presenting with globus symptoms. We conducted a retrospective analysis of 250 patients who underwent rigid endoscopy for globus symptoms over a 12-month period. In 217 patients (86.8 per cent) the examination of the larynx, pharynx and upper oesophagus was entirely normal. Abnormal findings included cricopharyngeal spasm in 12 patients (4.8 per cent), reflux in 11 (4.4 per cent), pharyngitis in two (0.8 per cent), web in two (0.8 per cent), and retention cyst in three (1.2 per cent). The 95 per cent confidence interval (CI) for the mean number of persons with malignancy based on the Poisson distribution is 0 and 3.7 (0 and 14.8 as rates per 1000). The relationship between the clinical diagnosis and endoscopic findings was examined using the chi-square test, with a p value of 0.0001. These results suggest that patients presenting with globus sensation are unlikely to harbour neoplastic lesions and therefore rigid endoscopy may well be an inappropriate investigation in this group. The risks, costs and discomfort associated with this intervention can often be avoided. PMID- 16359157 TI - Laryngeal tuberculosis following laryngeal carcinoma. AB - Laryngeal tuberculosis is a rare entity and the disease related to laryngeal cancer is extremely rare. We describe a case of laryngeal tuberculosis in a 74 year-old man with a history of radiotherapy for laryngeal carcinoma four months earlier. Laryngoscopy demonstrated a white mass on the right vocal fold at the site carcinoma had previously occupied. Recurrence of the cancer was suspected, but the biopsy result showed histological features of tuberculosis. We discuss the derangement of the host's mucosal barrier by the malignancy as a contributing factor in secondary tuberculous infection. Tubercular bacilli may be reactivated due to the immunosuppression associated with the therapy. PMID- 16359158 TI - Congenital epulis. AB - Congenital epulis or gingival granular cell tumour is a rare lesion occurring in the newborn, affecting females eight times more than males and arising more commonly from the maxillary than the mandibular alveolar ridge. We report the case of a five-day-old girl who presented to us with a large mass arising from the gingival mucosa of the mandible, causing feeding difficulty. Immediate surgical excision was followed by an uneventful recovery. The case is reported to make clinicians aware of this uncommon but easily treatable condition. PMID- 16359159 TI - Steroids in rhinoplasty: a survey of current UK otolaryngologists' practice. AB - The use of steroids during rhinoplasty to reduce post-operative periorbital oedema and ecchymosis has been advocated. A number of randomized controlled trials have demonstrated the benefit of steroids in rhinoplasty. The aim of this study was to determine current UK practice in the use of steroids during rhinoplasty performed by otolaryngologists. A postal survey of consultant otolaryngologists in the UK was conducted. We received 203 responses, with 115 consultants performing 12 or more rhinoplasties per year. Only 28 consultants (24 per cent) used steroids routinely in patients undergoing rhinoplasty and of these 11 used a protocol, although this was unpublished. Dexamethasone was the most common steroid used (82 per cent), being administered as a single intravenous dose of 8 mg in the majority of cases (54 per cent). There was no correlation between the use of steroids and the number of rhinoplasties performed by individual consultants. Despite the evidence supporting the use of steroids to reduce post-operative sequelae following rhinoplasty, only a minority of consultants in the UK appear to use them as part of their practice. PMID- 16359160 TI - Hydrostatic balloon dilatation of pharyngeal stricture under local anaesthetic. AB - Balloon dilatation is well established in the management of enteric strictures. The use of this technique in the pharynx has been reported under radiological and endoscopic guidance. We describe the hydrostatic dilatation of a benign pharyngeal stricture in a laryngectomy patient under local anaesthetic, without radiological guidance, in the ENT out-patient department. This procedure was effective and well tolerated in a patient who required regular dilatations. PMID- 16359161 TI - Chemistry of ice surfaces. Elementary reaction steps on ice studied by reactive ion scattering. AB - This Account describes a recent study of reactions on ice surfaces with the emphasis on the mechanistic features of elementary reactions steps. Cs(+) reactive ion scattering (Cs(+) RIS) and low-energy sputtering (LES) techniques monitor the reactions by detecting the molecules and ions on the ice surface. The types of reactions include molecule diffusion and migration, proton transfer, and some simple reactions on frozen water and alcohol surfaces. Ice surface reactions exhibit unique behaviors due to a kinetic constraint, resulting in the isolation of reaction intermediates, preferential stabilization of charged species, and diversity of reaction products. PMID- 16359162 TI - Molecules in carbon nanotubes. AB - This Account focuses on structural and dynamic behavior of molecules encapsulated in carbon nanotubes. The impact of the confinement on the molecular packing, orientation, translation, rotation, and reactivity is demonstrated for a range of fullerene and nonfullerene molecules. These phenomena are described and analyzed using the current understanding of molecule-nanotube and intermolecular interactions. PMID- 16359163 TI - Native state kinetic stabilization as a strategy to ameliorate protein misfolding diseases: a focus on the transthyretin amyloidoses. AB - Small molecule-mediated protein stabilization inside or outside of the cell is a promising strategy to treat protein misfolding/misassembly diseases. Herein we focus on the transthyretin (TTR) amyloidoses and demonstrate that preferential ligand binding to and stabilization of the native state over the dissociative transition state raises the kinetic barrier of dissociation (rate-limiting for amyloidogenesis), slowing and in many cases preventing TTR amyloid fibril formation. Since T119M-TTR subunit incorporation into tetramers otherwise composed of disease-associated subunits also imparts kinetic stability on the tetramer and ameliorates amyloidosis in humans, it is likely that small molecule mediated native state kinetic stabilization will also alleviate TTR amyloidoses. PMID- 16359164 TI - Fluctuating enzymes: lessons from single-molecule studies. AB - Recent single-molecule enzymology measurements with improved statistics have demonstrated that a single enzyme molecule exhibits large temporal fluctuations of the turnover rate constant at a broad range of time scales (from 1 ms to 100 s). The rate constant fluctuations, termed as dynamic disorder, are associated with fluctuations of the protein conformations observed on the same time scales. We discuss the unique information extractable from these experiments and the reconciliation of these observations with ensemble-averaged Michaelis-Menten equation. A theoretical model based on the generalized Langevin equation (GLE) treatment of Kramers' barrier crossing problem for chemical reactions accounts naturally for the observation of dynamic disorder and highly dispersed kinetics. PMID- 16359165 TI - Chemomechanical functionalization and patterning of silicon. AB - The chemomechanical method has emerged as a straightforward and convenient tool for simultaneously functionalizing and patterning silicon. This technique simply consists of wetting (or exposing) a silicon surface to a reactive chemical and then scribing. Scribing activates the surface and leads to monolayer formation. The properties of the monolayers are dependent on the reactive chemicals used, and mixed monolayers and funtionalized monolayers are easily produced with mixed chemicals or alpha,omega-bifunctional compounds, respectively. Both micrometer and nanometer sized functionalized features have been created. It has been shown that this technique has potential in a variety of applications. PMID- 16359166 TI - Metalloporphyrin-NO bonding: building bridges with organometallic chemistry. AB - DFT calculations in our laboratory and elsewhere have elucidated fundamental aspects of the structure and bonding of a variety of metalloporphyrin-diatomic complexes, including the biologically important heme CO, NO, and O(2) complexes. We have also studied some more exotic species such as metalloporphyrin dinitrosyl, -dialkyl, and -diaryl complexes. In the course of this research, we discovered a number of unexpected similarities (isolobal analogies) between the bonding in metalloporphyrin-NO and organometallic compounds. Equally important, DFT calculations have played a significant role in advancing our understanding of selective diatomic ligand binding by heme proteins. PMID- 16359167 TI - Catalytic reduction of dinitrogen to ammonia at a single molybdenum center. AB - This account explores the catalytic reduction of dinitrogen by molybdenum complexes that contain the [HIPTN(3)N](3-) ligand ([HIPTN(3)N](3-)) = [(HIPTNCH(2)CH(2))(3)N](3-), where HIPT = 3,5-(2,4,6-i-Pr(3)C(6)H(2))(2)C(6)H3) at room temperature and pressure with protons and electrons. A total of 7-8 equiv of ammonia is formed out of approximately 12 possible (depending upon the Mo derivative employed). No hydrazine is formed. Numerous X-ray studies of proposed intermediates in the catalytic cycle suggest that N(2) is being reduced at a sterically protected, single Mo center operating in oxidation states between Mo(III) and Mo(VI). Subtle variations of the [HIPTN(3)N](3-) ligand are not as successful as a consequence of an unknown shunt in the catalytic cycle that consumes reduction equivalents to yield (it is proposed) dihydrogen [corrected] PMID- 16359168 TI - The role of Keap1 in cellular protective responses. PMID- 16359169 TI - Identification of N-(hydroxymethyl) norcotinine as a major product of cytochrome P450 2A6, but not cytochrome P450 2A13-catalyzed cotinine metabolism. AB - Cotinine formation is the major pathway of nicotine metabolism in smokers, and the primary pathway of cotinine metabolism is trans-3'-hydroxylation. trans-3' Hydroxycotinine and its glucuronide conjugate account for up to 50% of the nicotine metabolites excreted by smokers. Minor metabolites of cotinine excreted by smokers include norcotinine and cotinine N-oxide, each of which account for <5% of the nicotine dose. It has been reported that P450 2A6 is the catalyst of cotinine metabolism. However, we report here that the major product of P450 2A6 catalyzed cotinine metabolism is N-(hydroxymethyl)norcotinine, a previously unknown human metabolite of cotinine. N-(Hydroxymethyl)norcotinine was chemically synthesized, and its stability under the conditions of the enzyme reactions was confirmed. The products of P450 2A6-catalyzed [5-3H]cotinine metabolism were quantified by radioflow HPLC. The identification of N-(hydroxymethyl)norcotinine as the major metabolite was based on HPLC analysis on three unique systems and coelution with N-(hydroxymethyl)norcotinine standard. 5'-Hydroxycotinine and trans-3'-hydroxycotinine were minor products of P450 2A6-catalyzed cotinine metabolism, accounting for 14 and 8% of the total cotinine metabolites, respectively. N-(Hydroxymethyl)norcotinine was a product of cotinine metabolism by the extrahepatic P450, 2A13, but it was a minor one. The major product of P450 2A13-catalyzed cotinine metabolism was 5'-hydroxycotinine, which was formed at twice the rate of trans-3'-hydroxycotinine. The identification of all cotinine metabolites formed by both enzymes was confirmed by LC/MS/MS analysis. Kinetic parameters for cotinine metabolism were determined for P450 2A6 and P450 2A13. This work has confirmed that the major metabolite of cotinine in smokers, trans 3'-hydroxycotinine, is only a minor metabolite of P450 2A6-catalyzed cotinine metabolism. PMID- 16359170 TI - Evidence of an immune-mediated mechanism for an idiosyncratic nevirapine-induced reaction in the female Brown Norway rat. AB - Previously, we reported a new animal model of an idiosyncratic drug reaction in which nevirapine causes a skin rash in some rats that has characteristics similar to the reaction that occurs in humans. Strong evidence that the reaction is immune-mediated was found; specifically, low-dose pretreatment induced tolerance, while with rechallenge, the time to onset decreased and the severity increased. Furthermore, splenocytes from rechallenged rats transferred rash susceptibility to naive recipients. We now report the results of studies to explore the immune aspects of this reaction. T cells were found to play an important role, as demonstrated by their ability to adoptively transfer susceptibility to the skin reaction. Of these T cells, CD4+ cells are the likely effectors because they were capable of transferring susceptibility and the reaction was delayed in rats partially depleted of CD4+ T cells. In contrast, it appears that CD8+ T cells are not essential, as CD8+ T cells were unable to transfer sensitivity to a naive animal and rats depleted of CD8+ T cells still developed skin rash. Unlike the penicillamine model, where we have demonstrated that the tolerance induced by low dose treatment is immune-mediated, tolerance induced by low-dose nevirapine appears to be largely due to induction of metabolism as it can be overcome by inhibition of cytochrome P450. Pretreatment with the immunosuppressants, cyclosporine and tacrolimus, prevented the rash and even led to resolution of the rash during nevirapine treatment. These studies reinforce the hypothesis that the reaction in this model is similar to that which occurs in humans. In particular, the finding that CD4+ T cells may play a central role in this model fits with the observation that the incidence of idiosyncratic reactions to nevirapine in humans appears to be lower in patients with low CD4+ counts. PMID- 16359171 TI - Visible study of mercuric ion and its conjugate in living cells of mammals and plants. AB - The investigation of mercuric ion and mercuric conjugate inside live specimens has drawn intense attention because of their cytotoxicity. The translocation, transportation, and distribution of Hg2+ inside either mammals or plants, however, are still invisible due to the lack of favorable fluorescent molecular sensors for Hg2+. Here, two sensors, 2,6-bis(4'-peperazino-N' hydroxylethoxylethylene-1',8'-naphthalimide)dimethylpyridine and 2,6-bis(4' peperazino-N'-butyl-1',8'-naphthalimide)dimethylpyridine, which were composed of nitrogen atoms of 2,6-bis(aminomethyl)pyridine as the ion receptor and the donor of photoinduced electron transfer, were applied to the cultured mammalian cells and plant cells. Their membrane permeability, low toxicity, slow bleaching/fading, and high selectivity and sensitivity to Hg2+ in a live cell rendered them attractive to become real-time and real-space sensors. For the first time, the transportation of Hg2+ cation and Hg2+ conjugate of cysteine was observed with the help of a fluorescence microscope. The chloroplast location of Hg2+ in transgenic tobacco was also visible. PMID- 16359172 TI - Chromatographic separation and identification of products from the reaction of dimethylarsinic acid with hydrogen sulfide. AB - The reaction of dimethylarsinic acid (DMAV) with hydrogen sulfide (H2S) is of biological significance and may be implicated in the overall toxicity and carcinogenicity of arsenic. The course of the reaction in aqueous phase was monitored, and an initial product, dimethylthioarsinic acid, was observed by using LC-ICP-MS and LC-ESI-MS. Dimethylarsinous acid was observed as a minor product. A second slower-forming product was identified, and the electrospray mass chromatograms for this species produced ions at m/z 275, 171, and 137 in positive mode. To aid in the identification of this slower-forming product, crystalline standards of sodium dimethyldithioarsinate and dimethylarsino dimethyldithioarsinate were prepared and re-characterized by using improved spectroscopic and structural analysis techniques. An aqueous solution of sodium dimethyldithioarsinate produced a single major chromatographic peak that matched the retention time (7.6 min) of the slower-forming product and contained similar molecular ions at m/z 275, 171, and 137 via LC-ESI-MS. The dimethylarsino dimethyldithioarsinate standard produced four aqueous phase species one of which coeluted with the slower forming product. This coeluting peak also produced the identical ESI-MS ions as the slower-forming product of DMAV + H2S. ESI-MS/MS experiments conducted on sodium dimethyldithioarsinate in deuterated water produced molecular ions at m/z 276, 173, and 137. Subsequent collisionally activated dissociation (CAD) experiments on m/z 276 did not produce a product ion at m/z 173. These data indicate that two different species are present in solution, while NMR data indicate that only dimethyldithioarsinic acid exists in aqueous solutions. This discrepancy was investigated by conducting NMR studies on the acidic solution of sodium dimethyldithioarsinate after taking this solution to dryness. The resolubilized solution produced a proton NMR signal characteristic of dimethylarsino dimethyldithioarsinate. Therefore, it was concluded that the ESI-MS ion at m/z 275 associated with the slowly forming second reaction product and the sodium dimethyldithioarsinate compound is a product of the ESI desolvation process. PMID- 16359173 TI - Oxanosine is a substrate of adenosine deaminase. Implications for the quest for a toxicological marker for nitrosation activity. AB - Oxanosine 3r, 5-amino-3-beta-(d-ribofuranosyl)-3H-imidazo[4,5-d][1,3]oxazine-7 one, was isolated as a novel nucleoside antibiotic in 1981 from Streptomyces capreolus MG265-CF3. Oxanosine became relevant in toxicology in 1996 with the discovery that it is formed in nitrosative guanosine deamination. As part of studies of the mechanism of oxanosine formation, the synthesis was attempted of [7- 18O]oxanosine by enzymatic 16O/18O-exchange with adenosine deaminase (ADA) in analogy to the synthesis of [6- 18O]guanosine from 2-amino-6-chloropurine. Unexpectedly, it was discovered that the incubation of oxanosine 3r with ADA in sodium phosphate buffer (pH = 7.4) results in 1-beta-(d-ribofuranosyl)-5-ureido 1H-imidazole-4-carboxylic acid 4r. The reaction of the 2'-deoxyribose derivative 3d forms 4d in analogy. The reaction products were separated by preparative RP HPLC and characterized by LC/MS and MS/MS analyses and UV/vis and NMR spectroscopy, and NMR assignments were corroborated by GIAO and GIAO-PCM calculations. Reaction in H2 18O leads to 18O-incorporation at C7. The hydrolysis of 3 to 4 can be rationalized on the basis of the known mode of action of ADA, and an explanation is provided for ADA's accomplishment of the "usual" substitution at C6 of adenosine (addition to the exocyclic bond) and the "lactone hydrolysis" of oxanosine (addition to the endocyclic double bond). The Michaelis Menten constant of Km = 1.0 (+/-0.2) mM was measured for oxanosine. Implications are discussed for studies of nitrosative deamination of nucleosides, nucleotides, and oligonucleotides. PMID- 16359174 TI - Stability and comparative metabolism of selected felbamate metabolites and postulated fluorofelbamate metabolites by postmitochondrial suspensions. AB - Evidence has been presented suggesting that a reactive metabolite, 2 phenylpropenal (ATPAL), may be responsible for the toxicities observed during therapy with the antiepileptic drug felbamate (FBM). Formation of ATPAL from its unstable immediate precursor, 3-carbamoyl-2-phenylpropionaldedhyde (CBMA) requires the loss of the hydrogen atom at position 2 in the propane chain, and it has been postulated that substitution of this atom with fluorine would prevent the formation of ATPAL. On the basis of this hypothesis, 2-fluoro-2-phenyl-1,3 propanediol dicarbamate (F-FBM) was synthesized and is presently undergoing drug development. To test this hypothesis, we compared the metabolism by human liver postmitochondrial suspensions (S9) in vitro of selected FBM and postulated F-FBM metabolites leading to formation of CBMA or 3-carbamoyl-2-fluoro-2-phenyl propionaldehyde (F-CBMA). All S9 incubations included GSH as a trapping agent for any reactive metabolites formed. Our results indicated that, in phosphate buffer, pH 7.4, at 37 degrees C, the half-life for 4-hydroxy-5-phenyltetrahydro-1,3 oxazin-2-one (CCMF) was 2.8 and 3.6 h in the presence or absence of GSH, respectively; compared to 4-hydroxy-5-fluoro-5-phenyl-tetrahydro-1,3-oxazin-2-one (F-CCMF) which lost only 2.5% or 4.9% over 24 h under the same conditions. When incubated with S9 in the presence of the cofactor, NAD+, 2-phenyl-1,3-propanediol monocarbamate (MCF) was oxidized to CCMF which was further oxidized to 3 carbamoyl-2-phenylpropionic acid (CPPA). 2-Fluoro-2-phenyl-1,3-propanediol monocarbamate (F-MCF) under similar conditions was stable, and no metabolites were observed. When CCMF was incubated with S9 in the presence of NAD+ cofactor, oxidation to CPPA and reduction to MCF were observed. In addition, a new atropic acid GSH adduct (ATPA-GSH) was identified by mass spectrometry. When F-CCMF was incubated under the same conditions as CCMF, both reduced and oxidized metabolites, F-MCF and 3-carbamoyl-2-fluoro-2-phenylpropionic acid (F-CPPA), respectively, were formed but at significantly lower rates, and no GSH conjugates were identified. Our results support the hypothesis that F-FBM and F-CCMF are not metabolized by S9 in vitro to the known reactive FBM metabolite, ATPAL. PMID- 16359176 TI - A quantitative structure--activity relationship model for the intrinsic activity of uncouplers of oxidative phosphorylation. AB - A quantitative structure-activity relationship (QSAR) has been derived for the prediction of the activity of phenols in uncoupling oxidative and photophosphorylation. Twenty-one compounds with experimental data for uncoupling activity as well as for the acid dissociation constant, pKa, and for partitioning constants of the neutral and the charged species into model membranes were analyzed. From these measured data, the effective concentration in the membrane was derived, which allowed the study of the intrinsic activity of uncouplers within the membrane. A linear regression model for the intrinsic activity could be established using the following three descriptors: solvation free energies of the anions, an estimate for heterodimer formation describing transport processes, and pKa values describing the speciation of the phenols. In a next step, the aqueous effect concentrations were modeled by combining the model for the intrinsic uncoupling activity with descriptors accounting for the uptake into membranes. Results obtained with experimental membrane-water partitioning data were compared with the results obtained with experimental octanol-water partition coefficients, log Kow, and with calculated log Kow values. The properties of these different measures of lipophilicity were critically discussed. PMID- 16359175 TI - Quantitation of four guanine oxidation products from reaction of DNA with varying doses of peroxynitrite. AB - The oxidation products obtained from the reaction of peroxynitrite (ONOO-) with dG include-among others-8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), 2,2 diamino-4[(2-deoxy-beta-d-erythro-pentafuranosyl)amino]-5(2H)-oxazolone (oxazolone), spiroiminodihydantoin, and N1-(beta-d-erythro-pentofuranosyl)-5 guanidinohydantoin (guanidinohydantoin). In the present work, the formation of these products from the treatment of calf thymus DNA with varying amounts of ONOO was studied quantitatively in vitro. 13C-, 15N-labeled standards were synthesized for the nucleosides of interest, and calf thymus DNA was reacted with ONOO- and digested enzymatically down to the nucleoside level. Specific modifications in the DNA were measured by HPLC separation followed by electrospray ionization tandem mass spectrometric analysis in the selected reaction-monitoring mode. Artifacts of the above four oxidation products, arising from oxidation of dG and/or 8-oxodG during DNA digestion and subsequent workup, were evaluated with 7-15N-dG and/or stable-isotope-labeled 8-oxodG as internal standards. Levels of artifactual 8-oxodG were about 5/10(6) nucleosides. The artifacts of spiroiminodihydantoin and guanidinohydantoin, arising from 8-oxodG, were 3.7% and 0.6% of the measured 8-oxodG values, respectively. No artifacts of oxazolone were detected. 8-OxodG and oxazolone were formed dose-dependently in DNA treated with ONOO-, while the levels of spiroiminodihydantoin and guanidinohydantoin increased significantly at low ONOO- doses, and then dropped off at higher ONOO- doses. The complexity of these dose-response relationships is likely due to the dual role of peroxynitrite as both an oxidant and a nucleophile in competition with water. PMID- 16359177 TI - P450 2C18 catalyzes the metabolic bioactivation of phenytoin. AB - The safe clinical use of phenytoin (PHT) is compromised by a drug hypersensitivity reaction, hypothesized to be due to bioactivation of the drug to a protein-reactive metabolite. Previous studies have shown PHT is metabolized to the primary phenol metabolite, HPPH, then converted to a catechol which then autoxidizes to produce reactive quinone. PHT is known to be metabolized to HPPH by cytochromes P450 (P450s) 2C9 and 2C19 and then to the catechol by P450s 2C9, 2C19, 3A4, 3A5, and 3A7. However, the role of many poorly expressed or extrahepatic P450s in the metabolism and/or bioactivation of PHT is not known. The aim of this study was to assess the ability of other human P450s to catalyze PHT metabolism. P450 2C18 catalyzed the primary hydroxylation of PHT with a kcat (2.46 +/- 0.09 min-1) more than an order of magnitude higher than that of P450 2C9 (0.051 +/- 0.004 min-1) and P450 2C19 (0.054 +/- 0.002 min-1) and Km (45 +/- 5 microM) slightly greater than those of P450 2C9 (12 +/- 4 microM) and P450 2C19 (29 +/- 4 microM). P450 2C18 also efficiently catalyzed the secondary hydroxylation of PHT as well as covalent drug-protein adduct formation from both PHT and HPPH in vitro. While P450 2C18 is expressed poorly in the liver, significant expression has been reported in the skin. Thus, P450 2C18 may be important for the extrahepatic tissue-specific bioactivation of PHT in vivo. PMID- 16359178 TI - A computational study of carbocations from oxidized metabolites of dibenzo[a,h]acridine and their fluorinated and methylated derivatives. AB - In a model computational study aimed at understanding structure-reactivity relationships and substituent effects on carbocation stability in aza-polycyclic aromatic hydrocarbons, the epoxides, diol epoxides, and the dihydrodiols of dibenzo[a,h]acridine (DB[a,h]ACR) were studied by density functional theory at the B3LYP/6-31G level. Bay region carbocations were formed via the O-protonated epoxides in barrierless processes. Relative carbocation stabilities were determined in the gas phase and in water as solvent (polarized continuum model method). Charge delocalization modes in the resulting carbocations were deduced by gauge-independent atomic orbitals (GIAO) NMR (based on Delta delta13C values) and via the natural population analysis (NPA)-derived changes in charges. Although the solvent decreases the exothermicity of the epoxide ring-opening reactions due to greater stabilization of the reactants, relative reactivity trends remain the same. Whereas fluorine substitution at ring positions bearing significant positive charge leads to carbocation stabilization by fluorine p-pi back-bonding, fluorine substitution at a ring position that presented negative charge density in the unsubstituted compound leads to inductive destabilization. Methylated derivatives exhibit less sensitivity to substituent effects as compared to the fluorinated analogues. A bay region methyl group produces structural distortion, and this deviation from planarity destabilizes the epoxide, favoring ring opening. Relative energies, changes in NPA charges, and GIAO NMR data in the resulting "benzylic" carbocations are examined collectively and discussed, taking into account the available biological activity data on these compounds. PMID- 16359179 TI - Liquid chromatography-mass spectrometric analysis of urinary metabolites and their pattern recognition for the prediction of drug-induced hepatotoxicity. AB - Multivariate pattern recognition (PR) analysis combined with LC/MS was utilized to evaluate the feasibility of predicting chemical-induced toxicity in rats. Urine samples were collected from rats treated with vehicles or four hepatotoxins, alpha-naphthyl isothiocyanate (ANIT), carbontetrachloride (CCl4), acetaminophen, and diclofenac, and analyzed by HPLC coupled with electrospray mass spectrometry. Chromatographic data were normalized using modified Z score transformation with those of the control group, to remove the vehicle effects for a further enhanced multivariate analysis. The LC/MS-based profiles of the urine samples showed different levels of endogenous metabolites, which were characteristic of each hepatotoxin. In the principal component (PC) map of the urinary spectra from rats treated with ANIT, the metabolic trajectory moved away from the predose position, reaching a maximum separation at the 32-48 h time period. The metabolic profiles partially recovered to the basal conditions on 7 days postdose. A principal component analysis was performed on the urinary spectra of rats treated with the vehicles or four hepatotoxins. Each group formed a distinct and isolated cluster in the PC map, indicating drug-induced perturbation in the urine profiles. To construct mathematical models for predicting drug-induced hepatotoxicity, supervised analyses, such as linear discriminant analysis and soft independent modeling of class analogy with residual distance (SIMCA-RD), were performed. The SIMCA-RD showed high predictability, over 95%, in the results of cross-validation using the leaving one-out method. The developed LC/MS-PR approach might be a useful tool for the prediction of drug-induced hepatotoxicity and for the understanding of hepatotoxic mechanisms. PMID- 16359180 TI - The reactivity of the 5-hydroxy-5,6-dihydrothymidin-6-yl radical in oligodeoxyribonucleotides. AB - Hydroxyl radical can be induced from Fenton reaction or gamma radiolysis of water. It can add preferentially to the C5 position of thymidine to give the 5 hydroxy-5,6-dihydrothymidin-6-yl radical. In this report, we examined the independent generation and reactivity of this radical in oligodeoxyribonucleotides (ODNs). Our results showed that the major products originated from this radical in single- and double-stranded ODNs were thymidine glycol, 5-hydroxy-5,6-dihydrothymidine, thymidine, and abasic site lesion. A cross-linking lesion, where the C6 of 5-hydroxy-5,6-dihydrothymidine and the C8 of its neighboring guanine are covalently bonded, could be induced from the independently generated radical in dinucleoside monophosphate and trinucleoside diphosphates. The formation of this type of cross-link product in duplex ODNs from either the independently generated radical or gamma irradiation was near or below the detection limit of the LC-MS/MS method that we used. PMID- 16359181 TI - Formation of transient covalent protein and DNA adducts by quercetin in cells with and without oxidative enzyme activity. AB - This study investigates the role of cellular tyrosinase and/or peroxidase-like oxidative enzyme activity in the covalent binding of quercetin to glutathione, protein, and DNA, as well as the stability of quercetin DNA adducts in time. This was done by studying the formation of glutathionyl quercetin adducts in various in vitro models, and the covalent binding of radiolabeled quercetin to protein and DNA in cells with elevated peroxidase or tyrosinase levels and in cells devoid of nucleotide excision repair (NER). Cells with elevated tyrosinase or peroxidase levels contained approximately 2 times higher levels of covalent quercetin adducts than cells without detectable levels of these oxidative enzymes. However, this difference was smaller than expected based on the differences in tyrosinase and/or peroxidase levels, indicating that these types of oxidative enzyme activities do not play a major role in the cellular pro oxidant activity of quercetin. Furthermore, quercetin DNA adducts were of transient nature, independent of the presence of NER, suggesting chemical instability of the adducts. Whether this transient nature reflects real reversibility or formation of genotoxic, depurinated sites remains to be investigated at the molecular level. Together, these data indicate that formation of covalent quercetin adducts can be expected in all cells, independent of their oxidative enzyme levels, whereas the transient nature of the DNA adducts formed may limit or cause their ultimate biological impact. If the transient nature represents chemical reversibility of the adduct formation, it would provide a possible explanation for the apparent lack of in vivo carcinogenicity of this in vitro mutagen. Therefore, in vitro mutagenicity studies should focus more on the transient nature of DNA adducts responsible for the mutagenicity in vitro, since this transient nature of DNA adducts may play an essential role in whether the genotoxicity observed in vitro will have any impact in vivo. PMID- 16359182 TI - Identification of sensor cysteines in human Keap1 modified by the cancer chemopreventive agent sulforaphane. AB - The chemopreventive agent sulforaphane is an isothiocyanate derived from cruciferous vegetables. Sulforaphane exerts cancer chemopreventive effects by inducing antioxidant/electrophile response element (ARE)-regulated phase 2 enzyme and antioxidant genes through activation of the transcription factor nuclear factor-E2-related factor 2 (Nrf2), which is regulated by the thiol-rich sensor protein Kelch-like ECH-associated protein 1 (Keap1). Sulforaphane is an electrophile that can react with protein thiols to form thionoacyl adducts. We hypothesized that, like other electrophilic Nrf2 activators, sulforaphane activates this system through specific modifications of the Keap1 protein. However, thionoacyl adducts are labile to hydrolysis and transacylation reactions, which complicate the identification of the sulforaphane adduct sites on Keap1. In this study, we characterized the stability of sulforaphane thionoacyl adducts and developed a liquid chromatography-tandem mass spectrometry method to map labile sulforaphane adduct sites formed on Keap1 in vitro. Sulforaphane displays a distinctly different pattern of Keap1 modification than previously studied ARE inducers that modify Keap1 by alkylation. Sulforaphane modified Keap1 most readily in the Kelch domain, rather than in the central linker domain, which is targeted by previously characterized ARE inducers. Also, in contrast to previously studied ARE inducers and as reported recently [Zhang, et al. (2005) J. Biol. Chem. 280, 30091-30099], sulforaphane treatment in vivo does not lead to the accumulation of ubiquitinated Keap1. Our observations suggest a novel mechanism for Nrf2 stabilization by sulforaphane-Keap1 thionoacyl adduct formation. PMID- 16359183 TI - Metabolism of aminoguanidine, diaminoguanidine, and NG-amino-L-arginine by neuronal NO-synthase and covalent alteration of the heme prosthetic group. AB - It is established that aminoguanidine (AG), diaminoguanidine (DAG), and NG-amino l-arginine (NAA) are metabolism-based inactivators of the three major isoforms of nitric oxide synthase (NOS). In the case of neuronal NOS (nNOS), heme alteration is known to be a major cause of inactivation, although the exact mechanism by which this occurs is not well-understood. We show here by the use of LC/MS/MS techniques that AG, DAG, and NAA are metabolized by nNOS to products with corresponding mass ions at m/z of 45.2, 60.2, and 160.0, respectively. These results are consistent with the loss of a hydrazine moiety from each inactivator. These findings are confirmed by exact mass measurements and comparison to authentic standards in the case of the products for NAA and AG, respectively. Moreover, the major dissociable heme product that was formed during inactivation of nNOS by AG, DAG, and NAA had molecular ions at m/z 660.2, 675.2, and 775.3, respectively. These results are consistent with an adduct of heme and inactivator minus a hydrazine moiety. In support of this, MS/MS studies reveal a fragment ion of heme in each case. With the use of 14C-labeled heme, we also show that in the case of AG, the dissociable heme adduct accounts for approximately one-half of the heme that is altered. In addition, we employ a software-based differential metabolic profiling method by subtracting LC/MS data sets derived from samples that contained nNOS from those that did not contain the enzyme to search for products and substrates in complex reaction mixtures. The metabolic profiling method established in this study can be used as a general tool to search for substrates and products of enzyme systems, including the drug-metabolizing liver microsomal P450 cytochromes. We propose that the metabolism-based inactivation of nNOS by AG, DAG, and NAA occurs through oxidative removal of the hydrazine group and the formation of a radical intermediate that forms stable products after H atom abstraction or reacts with the heme prosthetic moiety and inactivates nNOS. PMID- 16359184 TI - Truncation, deamidation, and oxidation of histone H2B in cells cultured with nickel(II). AB - Molecular mechanisms of nickel-induced carcinogenesis include interactions of Ni(II) cations with histones. Previously, we demonstrated in vitro and in cells that Ni(II) cleaved off the -SHHKAKGK C-terminal motif of histone H2A. In the present study, Western blotting of histones isolated from rat and human cell lines, cultured for 3-5 days with 0.05-0.5 mM Ni(II), revealed time- and dose dependent appearance of a new band of histone H2B. This effect was also induced by Co(II), but not by Cu(II), Cd(II), and Zn(II). Mass spectrometry and amino acid sequencing of proteins from the new band allowed for identification of two derivatives of the major variant of histone H2B. The larger protein was histone H2B lacking 16 N-terminal amino acids. The smaller one was histone H2B which, in addition to being shortened at the N-terminus, had nine amino acids deleted from its C-terminus. At both termini, the truncation occurred between lysine and alanine in the two identical -KAVTK- repeats of histone H2B. Also, the truncated H2B proteins had their Q22 residues deamidated and M59 and M62 residues oxidized to sulfoxides, a signature of oxidative stress. The truncation did not concur with apoptosis. Its mechanism involved activation by Ni(II) treatment of specific nuclear proteolytic enzymes belonging to the calpain family. The terminal tails of core histones participate in structuring chromatin and regulating gene expression. Therefore, the observed truncation and other modifications of histone H2B may assist in Ni(II) carcinogenesis through epigenetic mechanisms. PMID- 16359185 TI - Mechanisms of aurothiomalate-Cys2His2 zinc finger interactions. AB - Zinc finger motifs are present in a wide variety of regulatory proteins and generally function as interaction modules between macromolecules. These functional interactions are controlled by mechanisms of zinc (Zn2+)-binding and release. Besides Zn2+ certain electrophilic metals can potentially react with zinc finger domains and lead to changes in the structure and function of those domains. In these studies, the Cys2His2 zinc finger was chosen as a model for understanding how the gold (I) (Au1+) drug, aurothiomalate (AuTM), interacts mechanistically with the Zn2+ coordination sphere. DNA binding assays were used to analyze functional interactions between AuTM and two model Cys2His2 zinc finger transcription factors, TFIIIA and Sp1; inhibition in the micromolar range of AuTM was observed in both cases. Electrospray ionization mass spectrometry (ESI-MS) was utilized to examine molecular interactions between AuTM and a zinc finger peptide modeled after the third finger of Sp1 (Sp1-3). These experiments demonstrated Au1+ ions can bind the zinc finger structure and trigger the release of the Zn2+ ion. Quantifying the ESI-MS data allowed for a relative affinity value between Zn2+ and Au1+ ions to be calculated and shows Au1+ has a 4-fold higher affinity for Sp1-3 than Zn2+. Mechanistic differences between Zn2+ and Au1+ binding to the model Sp1-3 zinc finger were analyzed at isotopic resolution, and the metal-coordination spheres were probed with small molecules (H+, hydrogen peroxide, glutathione disulfide, and iodoacetamide). Natural isotope cluster analysis suggested the presence of a metal-thiol bond in the Cys2His2 zinc finger structure. Metal exchange reactions between zinc fingers demonstrated Zn2+ ions exchanged more rapidly than Au1+ ions. Circular dichroism (CD) exhibited differences in the secondary structure of the Sp1-3 model peptide when binding Zn2+ or Au1+ ions. PMID- 16359186 TI - The carcinogenic significance of reactive intermediates derived from 3-acetoxy- and 5-acetoxy-2-hydroxy-N-nitrosomorpholine. AB - N-Nitroso-2-hydroxymorpholine (NHMOR), a relatively reactive metabolite of two potent carcinogens, N-nitrosodiethanolamine (NDELA) and N-nitrosomorpholine (NMOR), has been reported to not be carcinogenic. Two isomeric acetate esters of the alpha-hydroxynitrosamines expected to be produced from the cytochrome P450 mediated metabolism of NHMOR have been synthesized, and their hydrolytic decomposition products, hydrolysis rates, and deoxyguanosine (dG) reaction adducts have been determined. N-Nitroso-3-acetoxy-2-hydroxymorpholine was prepared in high yield from the reaction of N-nitroso-2,3-dehydromorpholine with dry peracetic acid in glacial acetic acid or by the reaction of its dimethyldioxirane-produced epoxide with glacial acetic acid. The hydrolysis of this alpha-acetoxynitrosamine gave acetaldehyde (10%), ethylene glycol (55%), glyoxal (95%), and acetic acid. The pH rate profile for the hydrolysis of this nitrosamine was abnormal in that it exhibited pronounced base-catalyzed hydrolysis beginning at pH 5. The mechanism of hydrolytic decomposition is proposed to involve neighboring group participation with the formation of a reactive epoxide intermediate. N-Nitroso-3-acetoxy-2-hydroxymorpholine reacted with dG to give these guanine adducts after acidic deglycosylation: 1,N2-glyoxal (65%), 7-(2-hydroxyethyl)guanine (9%), and O6-hydroxyethylguanine (3%). N-Nitroso 5-acetoxy-2-hydroxymorpholine was synthesized from 2-hydroxyethylvinylnitrosamine by its oxidative conversion to the corresponding aldehyde followed by reaction with dry peracetic acid in glacial acetic. The hydrolytic decomposition products of this nitrosamine were 2-acetoxyacetaldehyde (65%), a rearrangement product, glycol aldehyde (15%), a trace of glyoxal, and acetic acid. The pH rate profile for the hydrolysis of this acetate is similar to other alpha-acetoxynitrosamines in that it exhibits a pH-independent region which gives way to base-catalyzed ester hydrolysis beginning at pH 7. The lower pH ( approximately 7 < 9) onset of base catalysis is proposed to involve base-catalyzed opening of the hemiacetal and intramolecular acyl transfer to give an unstable alpha-hydroxynitrosamine. N Nitroso-5-acetoxy-2-hydroxymorpholine was less reactive toward dG and gave the 1,N2-etheno-dG adduct (44%). The products from both of the isomeric alpha-acetoxy nitrosamines were judged to arise from diazonium ions produced from unstable alpha-hydroxynitrosamine intermediates. The high yield of the rearrangement product 2-acetoxyacetaldehyde could explain the low carcinogenic potential of NHMOR if it is mainly alpha-hydroxylated at the 5 carbon. Hydroxylation of NHMOR at carbon 3 is expected to yield a carcinogenic outcome. PMID- 16359189 TI - Characterization of a cmeABC operon in a quinolone-resistant Campylobacter coli isolate of Irish origin. AB - In this study, a tripartite-operon-encoding efflux system together with its regulatory gene was characterized in an Irish Campylobacter coli isolate CIT-382 showing high-level resistance to nalidixic acid and ciprofloxacin. Sequence comparisons revealed significant homology between C. coli and the cmeABC operon of Campylobacter jejuni. Conservation of functional sequence domains and motifs were noted among C. coli and similar operons in unrelated organisms. A transcriptional regulatory gene cmeR located proximal to cmeABC was also identified. C. coli CIT-382 harbored the Thr-86-Ile amino acid substitution in the gyrA gene. Accumulation studies with ethidium bromide in the presence of known efflux pump inhibitors confirmed the presence of efflux pump activity in C. coli CIT-382. The efflux pump inhibitor PAbetaN had no effect on the MICs to quinolones. Our data suggest that the gyrA gene mutation is the main contributor to the high-level nalidixic acid and ciprofloxacin resistance observed in this Irish C. coli CIT-382 isolate. PMID- 16359188 TI - Chronic heart failure beyond city limits. AB - INTRODUCTION: Chronic heart failure (CHF) develops in frail elderly individuals who have suffered an acute or sustained insult to the structural efficiency of the heart due to the presence of underlying heart disease and/or hypertension. It is also more common in individuals with disproportionately high levels of cardiac disease or its risk factors, for example lower socioeconomic status. As such, this epidemic is particularly significant for older people, males and Aboriginal people; groups who comprise a greater proportion of the population in rural and remote Australia. The aim of this study is to determine if the rates of CHF differ between urban and rural Australia. METHOD: CHF prevalence rates derived from well validated international CHF prevalence data were applied to the Australian Bureau of Statistics Census data for 2001 and weighted to reflect the proportion of Aboriginal people in each geographical stratum. RESULTS: Australia wide, the estimated prevalence of CHF was 17.87 per 1000, ranging from 13.98/1000 in the Australian Capital Territory to 29.50/1000 in rural Northern Territory. Overall, CHF was more prevalent in rural and remote regions (19.84/1000) and large urban centres (19.01/1000) than in capital cities (16.94/1000) (p<0.001). High prevalence rates were also noted in the idyllic rural locations favoured by retirees. In Victoria, Western Australia, South Australia and the Australian Capital Territory over 70% of the estimated individual cases were located in capital cities. In New South Wales, Queensland, Tasmania and the Northern Territory the highest proportion of cases occurred outside capital cities. CONCLUSION: The main significance of these findings is that while a majority of heart failure may occur among people living in cities (because that is where most people live), a disproportionate number of cases occur among people living outside these cities (due to age and other socio-demographic risk factors) where services may be fewer and less accessible. PMID- 16359190 TI - Natural history of drug-resistant clones of Streptococcus pneumoniae colonizing healthy children in Portugal. AB - A total of 3,539 Streptococcus pneumoniae (Pn) were recovered from 4,969 nasopharyngeal samples of children attending 13 day-care centers (DCCs) located in Lisbon, Portugal, during a surveillance study from January, 2001, through March, 2003, integrated in the European intervention project (EURIS, European Resistance Intervention Study). All Pn isolates were tested for anti-biotyping and drug-resistant pneumococci (DRPn) were further tested by serotyping and pulsed-field gel electrophoresis (PFGE). Overall carriage of Pn was very high (71.2%) and 39.9% of the isolates were resistant to antimicrobials (22.5% with decreased susceptibility to penicillin and 17.4% susceptible to penicillin and resistant to other antimicrobials). Serotypes 6B, 14, 23 F, 19F, and 19 A were prevalent among the 1,287 DRPn and 5.8% of the isolates were non-typeable. Eighty PFGE patterns were identified among 1,285 DRPn, and 93.1% of the DRPn belonged to 26 major clonal types that comprised: Pneumococcal Molecular Epidemiology Network (PMEN) clones (76.3%), Portuguese (PT)-DCC clones, previously detected in 1996 1999 (14.3%), and EURIS PT-DCC new clones, identified for the first time in the EURIS study, during 2001-2003 (9.4%). Comparing with previous Portuguese surveillance studies carried out since 1996, we observed that carriage increased from 47% to 71%, but no major changes were detected on the prevalence of pneumococcal serotypes. Moreover, although PMEN clones were predominant in all DCCs, in the present study the majority of them were gradually decreasing in time whereas several PT-DCC and new clones seemed to be increasing. PMID- 16359191 TI - Antimicrobial resistance data on 16,756 Streptococcus pneumoniae isolates in 1999: A Pan-Regional Multicenter Surveillance Study in France. AB - The rising prevalence of antibiotic-resistant Streptococcus pneumoniae is a phenomenon observed to different degrees around the world. The present national surveillance study report analyzes a total of 16,756 strains of S. pneumoniae collected across France in 1999. The overall prevalence of S. pneumoniae with decreased susceptibility to penicillin was 44%, to amoxicillin 26%, and to cefotaxime 17%. The proportion of high-level resistant strains to penicillin (MIC > 1 mg/L), amoxicillin and cefotaxime (MIC > 2 mg/L) remained low: 12.3%, 1.8%, and 0.4% respectively. Prevalence of resistance to other antibiotics was high: 53% to erythromycin, 41.7% to cotrimoxazole, 31.8% to tetracycline, and 24.6% to chloramphenicol. Prevalence of penicillin-resistant S. pneumoniae varied according to subject age and specimen source. It was higher in children (52.7%) than in adults (39.8%) and higher in strains isolated from middle ear fluid (63.6%) than from blood cultures (41.8%) in children. S. pneumoniae resistant to other antibiotics were more common in children than in adults, although figures showed geographical variations. Comparison with a previous study realized in 1997 in the same regions confirms a rising trend in the prevalence of resistant bacteria. Therefore, we conclude that prevalence of antibiotic-resistant S. pneumoniae in 1999 continued to rise in France, although strains with high-level resistance to penicillin remained stable. PMID- 16359192 TI - In vitro activities of tigecycline, ertapenem, isepamicin, and other antimicrobial agents against clinically isolated organisms in Taiwan. AB - This study evaluated the in vitro activities of tigecycline, ertapenem, isepamicin, and other comparators against 861 bacterial isolates recovered from patients treated in three major teaching hospitals in 2003. MICs to antimicrobial agents were determined by the agar dilution method. High rates of oxacillin resistance (58%) in Staphylococcus aureus (60 isolates), and vancomycin resistance (21%) and quinupristin-dalfopristin non-susceptibility (39%) in Enterococcus faecium (34 isolates) were found. Carbapenems had excellent in vitro activities (>or=98% susceptibility) against the 419 isolates of Enterobacteriaceae, with the MIC(50) and MIC(90) of imipenem, meropenem, and ertapenem being 0.25 and 4 mg/L, 0.03 and 0.12 mg/L, and 0.03 and 0.5 mg/L, respectively. For, Pseudomonas aeruginosa (74 isolates) and Burkholderia cepacia (21 isolates), meropenem (MIC(90), 0.25, 2, and 4 mg/L, respectively) had better in vitro activities than imipenem (MIC(90), 8, 4, and 32 mg/L, respectively) and ertapenem (MIC(90), 0.5, >32, and 32 mg/L, respectively). Isepamicin had a similar activity with amikacin against all Enterobacteriaceae, Pseudomonas aeruginosa, B. cepacia, and Acinetobacter baumannii, except for C. freundii isolates in which isepamicin had an eight-fold activity better than amikacin. Tigecycline had excellent in vitro activities against all isolates tested (MIC(90), 25 mm, susceptible for susceptibility of E. coli to trimethoprim/sulfamethoxazole. Thus, this approach enabled determination of the most accurate breakpoints that best fitted the specific prevalence of susceptibility in Israel. Thus, we suggest its adoption by microbiology diagnostic laboratories for the provision of accurate antimicrobial susceptibility results when using the disk diffusion test. PMID- 16359194 TI - CTX-M-1 extended-spectrum beta-lactamase-producing Proteus mirabilis in Greece. AB - To examine the dissemination of CTX-M-type extended-spectrum beta-lactamases (ESBLs) in clinical isolates of Proteus mirabilis, 91 nonrepetitive ESBL producing P. mirabilis were collected from infected patients in a tertiary Greek hospital during September, 2001, to May, 2004. A bla (CTX-M) gene was amplified in one isolate (strain A328), but bla (CTX-M) was not detected in any of the remaining ESBL producers. Sequencing results showed that P. mirabilis A328 produced a CTX-M-1 enzyme while PCR mapping of the genetic element carrying bla (CTX-M-1) revealed that the gene was located downstream of an ISEcp1B element. The cefotaxime resistance determinant was easily transferable and carried on a 70 kb plasmid. The emergence of CTX-M-1- producing P. mirabilis indicates the need for early recognition of such strains to be able to control their spread in our hospital and community environment. PMID- 16359195 TI - Increasing prevalence of imipenem-resistant Pseudomonas aeruginosa and molecular typing of metallo-beta-lactamase producers in a Korean hospital. AB - The types of metallo-beta-lactamases (MBLs), integrons, and genetic relatedness among Pseudomonas aeruginosa were investigated with a recent high prevalence of imipenem resistance in a Korean hospital. During 2000-2003, a total of 116 non duplicate imipenem-resistant P. aeruginosa isolates were analyzed by PCR and DNA sequencing to detect of bla (IMP-1), bla (VIM-1), bla (VIM-2), bla (SPM-1), intI 1, intI 2, and intI 3 genes. Among them, MBL-producing isolates were evaluated for genetic relatedness using pulsed-field gel electrophoresis (PFGE) profiles. Of 116 isolates, 21 (18.1%) carried bla (VIM-2) gene with the intI 1 gene. Analysis of VIM-2 procuders by PFGE grouped 21 isolates into eight different clusters. Six of eight cluster I strains, all of four cluster II strains, and all of three cluster III strains were isolated in 2000, 2002, and 2003, respectively. Data concluded that P. aeruginosa carrying bla (VIM-2) with a class 1 integron was the only type among MBLs. A hospital outbreak by VIM-2 producers occurred annually, which could be at least a part of a recent high prevalence of imipenem resistance. PMID- 16359196 TI - Group A Streptococci from carriage and disease in Portugal: evolution of antimicrobial resistance and T antigenic types during 2000-2002. AB - In this study, we analyzed the antimicrobial resistance properties and T antigenic types of 511 isolates collected in Lisbon district, Portugal, from throat swabs of healthy subjects (n=341), during 2000-2002 and from diverse infection sites (n=170) of outpatients and inpatients, during 1999-2002. Erythromycin resistance was higher in tonsillitis/pharyngitis (27.4%) and skin infection isolates (21.1%), than in carriage and invasive isolates (or= 64 microg/ml olaquindox. In nine of the ten strains, the oqxA gene was detected. Sequencing of an internal fragment of oqxA from the oqxA-positive strains showed no variation, indicating highly conserved oqxA genes. All of the oqxA-positive strains contain plasmids with replicons similar to that of pOLA52. It was verified by Southern hybridization that the oqxAB operon was situated on plasmids in most, if not all, resistant strains. Furthermore, horizontal transfer of olaquindox resistance from three olaquindox resistant isolates was achieved using an olaquindox-sensitive E. coli as recipient. PMID- 16359199 TI - Class 1 integrons in Enterobacteriaceae isolated from clinical infections of horses and dogs in the Netherlands. AB - Integrons in gentamicin- and cotrimoxazole-resistant Enterobacteriaceae from dogs and horses with clinical infections were analyzed by conserved segment PCR-RFLP. Five distinct integron types were found, most of which have previously been reported in Enterobacteriaceae isolated from humans and farm animals, indicating that resistance genes are exchanged between the reservoirs in humans, farm animals, and companion animals. PMID- 16359200 TI - Variability in acquired resistance of Pasteurella and Mannheimia isolates from the nasopharynx of calves, with particular reference to different herd types. AB - To measure the level of antimicrobial resistance in potential bovine respiratory pathogens at different production types, nasal swabs were collected from 57 calves of 13 dairy herds, 150 calves of 9 beef cattle herds, and 289 calves of 5 high-density veal calf herds and investigated for the presence of Pasteurellaceae. All calves were less than 6 months old. Susceptibilities of the Pasteurella and Mannheimia isolates to eight antimicrobials were determined using an agar dilution method. P. multocida (37.3%) and hemolytic Mannheimia organisms (M. haemolytica sensu lato) (6.3%) were the most frequently detected organisms. The overall prevalence of isolates resistant to at least one antimicrobial from the dairy, beef, and veal calves were 17.6% (6/34), 21.9% (14/64), and 71.9% (64/89), respectively. In isolates obtained on the veal calf herds, acquired resistance to ampicillin, oxytetracycline, potentiated sulfonamides, gentamicin, tilmicosin, and enrofloxacin was frequently present, and 32.6% of these isolates were resistant to more than two of the tested antimicrobials. Resistance to ceftiofur and florfenicol was not detected. A substantial within-herd variability of species diversity and resistance profiles among isolates belonging to the genera Pasteurella and Mannheimia was found among the isolates of the veal calf farms. PMID- 16359201 TI - Presence and mechanism of antimicrobial resistance among enterococci from cats and dogs. AB - The presence and mechanism of acquired resistance to erythromycin, tylosin, lincomycin, quinupristin/dalfopristin, tetracycline, chloramphenicol, gentamicin, kanamycin, and vancomycin were determined in 97 and 104 enterococci isolated from rectal swabs of cats and dogs, respectively. Eleven feline and three canine enterococcal isolates contained the aac(6')-Ie-aph(2'')-Ia gene encoding high level resistance to gentamicin, an antibiotic often used for treating enterococcal infections in humans. The combination of erm(B) and vat(E) genes encoding resistance to streptogramins was detected in one canine quinupristin/dalfopristin-resistant Enterococcus faecium isolate. Four quinupristin/dalfopristin-resistant enterococci only contained the erm(B) gene. Cross resistance against macrolides and lincosamides (30%) and resistance against tetracyclines (55%) was found to be widely distributed among enterococci from pets. In all of the feline and in 93% of the canine macrolide and lincosamide resistant isolates, this resistance was encoded by the erm(B) gene. tet(M) was the most prevalent tetracycline resistance gene. It was detected in 91% of the feline and 86% of the canine tetracycline- resistant enterococci. A high occurrence of the Tn916/Tn1545 transposon family was found among these tet(M) positive isolates. Enterococci from pet animals with resistance against vancomycin were not found. This study shows that enterococci from the intestinal microbiota of cats and dogs may act as a reservoir of resistance genes for animal or human pathogens. PMID- 16359202 TI - Update on laparoscopic nephrectomy and nephroureterectomy. PMID- 16359203 TI - The seminal contribution of Georg Kelling to laparoscopy. PMID- 16359204 TI - Sir William Osler's perceptions of urolithiasis and the case of the indigo calculus. AB - BACKGROUND: Sir William Osler published his textbook, The Principles and Practice of Medicine, in 1892. It became the definitive treatise on a wide variety of diseases. The section on nephrolithiasis clearly presents the etiology, pathology, symptoms, diagnosis, and treatments. What remains a mystery is the mention, under rare forms of human stones, of a type called "indigo." MATERIALS AND METHODS: A search of Index Medicus starting from 1909 backward to its inception in 1879 was performed for key words "indigo," "calculus," "renal" or "bladder stones" and "indicanuria." Twelve textbooks of urology published before 1940 were scrutinized for references to indigo calculi. RESULTS: Only two references to indigo were found, both related to its use for treating constipation (1887 and 1891). Of the 12 textbooks, only 4 make passing reference to "indigo stones." They all mention that such calculi are very rare, but direct references to cases are lacking. One textbook references a study of blue stones from Egyptian mummies. CONCLUSION: It is unlikely that Osler's reference to an indigo calculus was taken lightly during his writing of The Principles and Practice of Medicine. The case of the indigo calculus is fascinating and perhaps enlightening if only for the source of Osler's intrigue. PMID- 16359205 TI - Percutaneous nephrolithotomy in patients who previously underwent open nephrolithotomy. AB - BACKGROUND AND PURPOSE: Open stone surgery nowdays is rare. However, some patients who are treated today have in the past undergone open nephrolithotomy. The aim of this study was to determine the possible impact of open nephrolithotomy on the efficacy and morbidity of subsequent percutaneous nephrolithotomy (PCNL). PATIENTS AND METHODS: We reviewed the files of all 167 patients undergoing PCNL at our institution between December 2000 and December 2003. The same surgeon performed all of the procedures. We compared 21 patients undergoing PCNL after open nephrolithotomy to the same kidney with all other patients undergoing PCNL. The groups did not differ in terms of age or stone burden (mean size, number of stones, percentage with staghorn calculi). The outcomes measured were operating time, necessity for secondary procedures, stonefree rate, and intraoperative and postoperative complications. RESULTS: The operating time (203+/-92 v 177+/-52 minutes) and percentage of secondary procedures (29% v 12%) were significantly higher in patients who had previously undergone open stone surgery. However, the stone-free rate (95% v 93%), intraoperative complication rate (10% v 9%), and postoperative complication rate (10% v 11%) did not differ significantly. CONCLUSIONS: A PCNL in a patient with a history of open nephrolithotomy may take longer and lead to a higher percentage of auxiliary procedures, probably because of scar tissue and anatomic changes in the kidney. However, the morbidity and efficacy of PCNL appear to remain the same in these patients. PMID- 16359207 TI - Laparoscopic adrenalectomy: a single-center experience of 43 cases. AB - PURPOSE: To evaluate the surgical feasibility of laparoscopic adrenalectomy and what laparoscopy offers for the surgeon and the patient. PATIENTS AND METHODS: From March 1996 to June 2004, 43 transperitoneal laparoscopic adrenalectomies were performed for various pathological states. Functioning adrenal masses and solid masses>5 cm were the most common indications. The mean size of the masses on abdominal CT was 6.8 cm in the largest diameter. All patients were assessed regarding the operative time, blood loss, complications, and conversion to open surgery. The postoperative course was reported with special attention to the complications and hospital stay. RESULTS: The mean operative time was 125 minutes with a mean blood loss of 60 mL. Intraoperative complications occurred in 3 cases (6.9%), necessitating conversion to open surgery in 2 to control bleeding from the avulsed right adrenal vein. A third case of conversion was elective because of difficult dissection of a large left pheochromocytoma from the renal hilum, so there was a 6.9% rate of conversion to open surgery. All patients showed early ambulation, early start of eating, and a short hospital stay (mean 2.6 days). CONCLUSION: Laparoscopic adrenalectomy is surgically feasible and can be applied for different adrenal pathologies. The procedure can be performed with a reasonable operative time, minimal blood loss, and an acceptable rate of complications. Laparoscopic adrenalectomy provides excellent postoperative recovery and convalescence with a short hospital stay. PMID- 16359206 TI - Efficacy and safety of fast-track recovery strategy for patients undergoing laparoscopic nephrectomy. AB - BACKGROUND AND PURPOSE: Factors that adversely affect early recovery after major laparoscopic procedures include ileus, pain, nausea, emesis, and fatigue. The objective of this randomized controlled study was to evaluate the impact of a multimodal fast-track (FT) rehabilitation program on recovery and length of hospital stay after laparoscopic nephrectomy. PATIENTS AND METHODS: Thirty patients undergoing laparoscopic nephrectomy received either conventional care (control) or an FT recovery program. All patients received a standardized anesthetic technique and patient- controlled analgesia (morphine) for postoperative pain control. In the FT group, patients received premedication with rofecoxib and ranitidine, local anesthesia was administered at the ports and renal fosa during surgery, and postoperative non-opioid analgesic and gastrokinetic drugs were administered as part of an early enteral nutrition and mobilization program. During the postoperative period, pain and nausea were assessed at specific time intervals. In addition, recovery room and hospital discharge times, the need for rescue analgesics and antiemetics, patient satisfaction with pain management and quality of recovery, and side effects were recorded daily for 3 days after surgery. Patients were discharged home when they met previously defined discharge criteria. RESULTS: The FT group was discharged earlier from the recovery room (74+/-23 v 103+/-47 minutes) and the hospital (41+/-11 v 59+/-11 hours). Pain and nausea scores were consistently lower in the FT group during the first 48 hours after surgery. In addition, the requirement for antiemetic rescue therapy during the first 24 hours was reduced in the FT group (15% v 58%). The FT group also received less morphine during the first 2 postoperative days (14+/-16 v 40+/-24 mg). Finally, patient satisfaction with postoperative pain control was significantly higher in the FT group. CONCLUSIONS: A multimodal approach to minimizing postoperative side effects led to a reduced recovery room and hospital stay, as well as better pain control and patient satisfaction after laparoscopic nephrectomy. PMID- 16359208 TI - Laparoscopic repair of ureterovaginal fistula: successful outcome by laparoscopic ureteral reimplantation. AB - We describe our technique of laparoscopic repair of ureterovaginal fistula in a 34-year-old woman who developed a vaginal leak of urine 15 days after laparoscopic hysterectomy. Laparoscopy and extravesical ureteral reimplantation with a stent was carried out. Cystoscopy and stent removal was done after 6 weeks. The patient was dry. To our knowledge, laparoscopy for the repair of ureterovaginal fistula has not been described. This distressing condition can be corrected laparoscopically with obvious benefit to the patient. PMID- 16359209 TI - Endoscopic diagnosis and management of ureteral endometriosis. AB - It is estimated that 1% of patients with endometriosis have involvement of the urinary tract, with the bladder being the most common location. Ureteral endometriosis is a rare entity, and the majority of cases are found at exploratory laparotomy for extensive involvement of the pelvic organs. Obstruction of the ureter may be caused by extrinsic or intrinsic disease, with the extrinsic form occurring four times as often. Progressive ureteral obstruction can be insidious in onset and ultimately lead to renal failure. Hormone therapy has had variable success, and open surgery has been the mainstay of treatment. Only one case of ureteral endometriosis, both intrinsic and extrinsic, diagnosed at ureteroscopy has been reported previously. We present a case of ureteral obstruction secondary to isolated intrinsic endometriosis diagnosed at ureteroscopy and treated endoscopically with holmium laser ablation and leuprolide therapy. PMID- 16359210 TI - Laparoscopic urology training in South Africa. AB - BACKGROUND AND PURPOSE: Laparoscopic urologic surgery is not widely practiced in South Africa. After presenting a laparoscopic training course, we evaluated how effectively this training translated into clinical practice. SUBJECTS AND METHODS: Invitations to the course were sent to all South African urologists. Ten applicants attended the course, which consisted of dry and in-vivo animal surgery. Two questionnaires were sent out after the course. Questionnaire 1 (at course completion) aimed at identifying the precourse laparoscopic experience and expectations of the trainee. Questionnaire 2 (6 months postcourse) assessed how much laparoscopic surgery the participant had performed since the course. RESULTS: Seventeen percent of all South African urologists responded to the invitation. Prior to the course, 40% of trainees had performed >or=10 laparoscopic cases, 30% had performed <10 cases, and 30% had never performed laparoscopy, whereas 60% expected to be doing one or two cases a month after completing the course. Six months after the course, 60% had performed no laparoscopic cases. Of the three trainees who had never before done laparoscopic procedures, none had started to perform procedures since the course. The commonest procedures performed were varicocelectomy and diagnostic laparoscopy for nonpalpable testis. CONCLUSIONS: A hands-on laparoscopic training course to introduce laparoscopic urology into South African private urology practice has not translated into a satisfactory number of clinical cases being performed. The causes are likely multifactorial but are greatly influenced by social and economic forces. One possible solution may be to offer a mentor-based training program. PMID- 16359211 TI - Subureteral dextranomer/hyaluronic acid copolymer injection for vesicoureteral reflux in transplant candidates. AB - BACKGROUND AND PURPOSE: Endoscopic subureteral injection of tissue-augmenting substances has become an alternative to antibiotic prophylaxis and open surgery for the management of vesicoureteral reflux (VUR). Several injectable materials have been tried for this purpose. In this study, we tried to determine the efficacy of dextranomer/hyaluronic acid copolymer (Dx/HA) injection for the treatment of VUR in renal-transplant candidates. PATIENTS AND METHODS: A total of 21 transplant candidates (29 ureteral units; 13 females, 8 males) with a mean age of 20.2 years (range 14-26 years) underwent endoscopic correction of VUR with Dx/HA. Diagnosis of VUR was made by voiding cystourethrography. The efficacy of the treatment was assessed with voiding cystourethrography at 3 months and 1 year postoperatively. Renal transplantation with living related donor organs was performed in 11 of the 21 patients. RESULTS: Endoscopic treatment was performed without complication in all cases. Higher success rates were obtained in patients with low-grade reflux, the overall success rate in the series being 82.7%. The mean follow- up after renal transplantation was 21.8 months (range 5-45 months). In one patient, reflux recurred after renal transplantation and was treated successfully by a repeat Dx/HA injection. The urine cultures of all patients remained sterile. CONCLUSION: Transplant candidates with VUR can be treated with Dx/HA, which cured the majority of our patients after one or two treatments with few low side effects. Endoscopic subureteral injection of Dx/HA has become an alternative treatment for VUR in transplant candidates. Long-term results are needed before making a final statement about its value. PMID- 16359212 TI - Vas deferens endoscopy (vasoscopy): a new diagnostic tool? AB - PURPOSE: To assess whether an experimental endoscopic study of the vasa deferentia is feasible. PATIENTS AND METHODS: Endoscopic study of the seminal tracts was carried out during three vasectomies performed prior to open prostatectomy. A flexible endoscope, 0.56 mm in diameter, with a depth of field of 5 mm and a 3000 fiberoptic light source was used for the procedure. RESULTS: Endoscopy was possible for the inguinoscrotal segment of the vas deferens. The curvature at the opening of the internal inguinal ring proved impassable. CONCLUSIONS: Endoscopy of the vas deferens is possible with a 0.56-mm flexible endoscope, but because of the extreme curvature of the vas at the inguinal ring, the complete vas cannot be evaluated in an antegrade manner. PMID- 16359213 TI - Long-term results of microwave thermotherapy for symptomatic benign prostatic hyperplasia. AB - PURPOSE: To study the long-term outcomes of men with moderately severe symptomatic benign prostatic hyperplasia (BPH) who were treated with transurethral microwave thermotherapy (TUMT) with the Dornier Urowave machine. PATIENTS AND METHODS: A total of 220 patients (mean age 66.2 years) with clinical BPH, an American Urological Association (AUA) Symptom Score of >or=13, and a peak urinary flow rate (Qmax) of 6 months) robotic experience, and four had none. Drills were performed randomly in both dimensional modes in a blinded fashion. Drill 1 was an interrupted four stitch and drill 2 a running closure. All tasks were kept uniform. We recorded time to completion, difficulty, and accuracy. The drills were evaluated by two independent reviewers for accuracy and major errors (i.e., broken suture, torn graft). RESULTS: The average operative time per drill in two dimensions was 13.1 minutes (range 6.9 21.9 minutes) and in three dimensions was 8.5 minutes (range 4.7-12.8 minutes) (P<0.001). Drill 1 was 6.1 minutes faster in three dimensions (mean 9.2 minutes; P<0.01), and drill 2 was 2.9 minutes faster (mean 7.8 minutes; P=0.03). Both advanced and novice groups were faster in 3D (P<0.01). There were two major errors in the 3D performances and 5 in the 2D exercises (P<0.05). The participants correctly identified the dimensional mode 92.9% of the time (P<0.01). CONCLUSION: The anastomosis was completed 65% faster using 3D with equal, if not greater, accuracy. Drill 1 was improved to a greater degree than drill 2, suggesting most benefit of 3D views during knot tying. Use of three dimensions outperformed two dimensions in both groups. Surgeons can immediately benefit from 3D viewing during robotic surgery. PMID- 16359219 TI - Novel everting urologic access sheath: decreased axial forces during insertion. AB - BACKGROUND: Advancement of urologic instruments through the genitourinary tract is associated with significant axial forces that likely contribute to patient discomfort, even after injection of a local anesthetic, and may lead to mucosal trauma, postprocedural dysuria and hematuria, and increased susceptibility to infection and strictures. Placing an everting urethral sheath prior to instrumentation may decrease these problems. MATERIALS AND METHODS: Two 7-cm long, 5-mm diameter urethral luminal models were created, one with and one without an artificial stricture. We measured the forces generated during advancement of a novel everting access sheath (Cystoglide; Percutaneous Systems, Mountain View, CA) through the models in comparison with a representative cystoscope and a urologic dilator simulating a traditional access sheath. RESULTS: The mean force generated during advancement of the everting sheath was significantly less than that of both the representative cystoscope (P<0.01) and the traditional access sheath (P<0.01). This held true for the urethral models both with and without an artificial stricture (P<0.01) and with and without lubrication (P<0.01). CONCLUSIONS: This novel introduction sheath markedly decreased the axial forces applied to an artificial urethral luminal wall. It is possible that the clinical use of this technology will decrease the discomfort and potential complications associated with lower urinary-tract endoscopy. PMID- 16359220 TI - Comparison of holding strength of suture anchors for hepatic and renal parenchyma. AB - BACKGROUND AND PURPOSE: Various laparoscopic devices have been described for suture anchoring during solidorgan parenchymal closure. Application of these devices expedites the closure of parenchymal defects and minimizes ischemia time. We compared different technologies as suture anchors for parenchymal closure. MATERIALS AND METHODS: A tensometer was used to determine the amount of tension necessary to dislodge each of five different clips from Vicryl suture alone or against two different substrates (fresh pig kidney and liver) with and without an intervening pledget. The clips investigated were the Lapra-Ty (Ethicon), Endoclip II (US Surgical), small Horizon Ligating Clips (Weck), Hem-o-lok Medium Polymer Clips (Week), and a novel Suture-clip (Applied Medical). ANOVA and two-sided Fisher's exact test provided statistical analysis. RESULTS: The force required to dislodge the Lapra-Ty clip from bare suture for both 0 and 1 Vicryl (7.0 N) was approximately fourfold the force required to dislodge the Endoclips or the 5-mm or 10-mm Hem-o-lok clips (p<0.01). When clips were applied to suture running through renal or liver parenchyma, the novel Suture-clip required the greatest tension to dislodge (P<0.01), followed by the Horizon and Lapra-Ty clips. There were no statistically significant differences in the tension required to dislodge a given clip from the two parenchymal substrates or in the presence or absence of a pledget. CONCLUSIONS: In our experimental model, the Suture-clip, Lapra-Ty, and Horizon clips required significantly greater tension to dislodge than the Hem-o lok and Endoclip clips. The addition of a pledget did not improve tension resistance. PMID- 16359225 TI - Dendritic cells, toll-like receptors, and T-cell responses: lessons from viral infections in vivo. PMID- 16359226 TI - Role of mucosal immunity in preventing genital herpes infection. AB - The female genital tract is immunologically unique because it must be tolerant to spermatozoa, pregnancy, and vaginal flora, but also protect the host from pathogen challenge. The mucosal response to herpes simplex viruses (HSV), a major cause of genital ulcerative disease and critical co-factor in the HIV/AIDS epidemic, is complex and consists of the normally acidic vaginal environment, constitutively secreted and induced antimicrobial peptides, complement, and cellular responses mediated by epithelial and immune cells. This review summarizes our current understanding of the mucosal response to HSV focusing on those factors that may prevent initial infection. Understanding how each of these components contribute to innate immunity, mechanisms of antiviral activity, and whether the virus has evolved strategies to evade their effects may lead to the development of novel vaginal microbicides. PMID- 16359227 TI - Rat cytomegalovirus induces a temporal downregulation of major histocompatibility complex class I cell surface expression. AB - Herpesviruses are known to influence expression of major histocompatibility complex (MHC) class I molecules on the surface of infected cells using a variety of mechanisms. Downregulation of MHC class I expression prohibits detection and elimination of infected cells by cytotoxic T lymphocytes. To investigate the effect of rat cytomegalovirus (RCMV) infection on MHC class I expression, we infected immortalized and primary rat fibroblasts with RCMV and monitored surface expression of MHC class I molecules at various time-points postinfection. These experiments revealed a downregulation of MHC class I surface expression by RCMV, a phenomenon that has also been reported for human and murine CMV. However, in contrast to the other cytomegaloviruses, RCMV causes only a temporal downregulation of MHC class I, with a maximal decrease at 12 h postinfection. Unlike murine and human CMV, RCMV does not induce proteolytic degradation of MHC class I molecules. In RCMV-infected cells, the MHC class I molecules are stable, but their exit from the ER is delayed. PMID- 16359228 TI - CD8+ T-cells suppress antigen-specific and allogeneic CD4+ T-cell responses to herpes simplex virus type 2-infected human dendritic cells. AB - In this study we show that human dendritic cells (DC), productively infected with herpes simplex virus type 2 (HSV-2), activate CD8+ T cells that suppress antigen specific and alloreactive CD4+ T cell expansion. Addition of CD8+ T cells to cultures of DC and CD4+ T cells blocked CD4+ T-cell proliferation in response to HSV-2-infected but not to uninfected DC. The effect was independent of prior HSV exposure or cognate MHC class I-restricted CD8-DC recognition as it was induced in CD8+ T cells from HSV-2-seronegative individuals and in mixed lymphocyte reactions using allogeneic DC. Both CD8+ CD25+ and CD8+ CD25- cells were shown to have suppressive capacities. The blood-derived CD25+ CD8+ T cells did not express Foxp3 mRNA but had a bona fide antiproliferative capacity in response to both uninfected and HSV-2-infected DC, whereas the CD25-CD8+ T cells were selectively activated to become antiproliferative by HSV-2-infected DC. These data imply that HSV infection of DC could modulate the immune response by activating CD8+ T cells. PMID- 16359229 TI - Recognition patterns of HLA-A2-restricted human immunodeficiency virus-1-specific cytotoxic T-lymphocytes in a cohort of HIV-1-infected individuals. AB - The cytotoxic T-lymphocytes (CTL) response to three histocompatibility leukocyte antigen (HLA)-A2-restricted CTL epitopes was investigated in a cohort of 51 HLA A2-positive human immunodeficiency-1 (HIV-1)-infected subjects. CTL activity was evaluated by testing peptide stimulated peripheral blood mononuclear cells (PBMC) in chromium release assays. The most prevalent CTL response was directed to the RT-peptide ILKEPVHGV (IV9) recognized by 37.3%. The p17-peptide SLYNTVATL (SL9), reported to be the immunodominant epitope in chronically infected untreated patients, was recognized only by 13.7%. Only 9.8% recognized both IV9 and SL9, and none recognized the RT-peptide VIYQYMDDL (VL9). CTL activity correlated significantly with absolute CD8 T-cell counts but not with CD4 counts, viral load, or antiviral therapy. Analysis of the recognition patterns of amino acid substitutions in the IV9 epitope revealed the presence of at least four functionally different T-cell receptors (TCR) in this cohort. All analyzed mutations within the TCR recognition site of this epitope could abrogate CTL recognition by individual CTL clones, but all were fully immunogenic for other CTL clones with peptide-sensitizing capacities similar to that of IV9. Further studies should be performed to evaluate whether a convergent epitope vaccination strategy using immunogenic variants of CTL epitopes is a feasible approach to broaden the TCR repertoire and to inhibit CTL escape. PMID- 16359230 TI - Comparison of antibody- and cell-mediated immune responses after intramuscular hepatitis C immunizations of BALB/c mice. AB - Current treatments for hepatitis C infection have limited efficacy, and there is no vaccine available. The goal of this study was to compare the immune response to several immunization combinations against hepatitis C virus (HCV). Six groups of mice were immunized at weeks 0, 4, and 8 with different combinations of a candidate HCV vaccine consisting of 100 microg recombinant HCV core/E1/E2 (rHCV) DNA plasmid and/or 25 microg rHCV polyprotein and 50 microL Montanide ISA- 51. Four weeks after the last injection, all groups of mice were sacrificed and blood samples and spleens were collected for measuring the levels of specific HCV antibodies (total IgG, IgG1, and IgG2a). Cell proliferation and intracellular interferon-gamma were also measured. Among the groups of immunized mice, only the mice immunized with rHCV DNA plasmid, rHCV polyprotein, and montanide (group D) and mice immunized with rHCV polyprotein and montanide (group F) demonstrated a significant increase in the total IgG titer after immunization. IgG1 was the predominant antibody detected in both groups D and F. No IgG2a was detected in any of the groups. Proliferation assays demonstrated that splenocytes from group D and group C (rHCV DNA primed/rHCV polyprotein boost) developed significant anti HCV proliferative responses. The combination of an rHCV DNA plasmid, rHCV polyprotein, and montanide induced a high antibody titer with a predominance of IgG1 antibodies and recognized the major neutralization epitopes in HVR1. In contrast, group C did not show an increase in anti-HCV antibodies, but did show a proliferative response. PMID- 16359231 TI - Adjuvant action of murine IL-2/Ig plasmid after intramuscular immunization with Indian HIV-1 subtype C recombinant env.gp 120 construct. AB - The human immunodeficiency virus (HIV) epidemic is probably the greatest scourge to affect mankind in the 20th century. Containment of the acquired immunodeficiency syndrome (AIDS) epidemic will require an effective vaccine. Of various vaccine approaches, immunization with DNA plasmids containing HIV-1 structural genes is the most popular approach. However, an important limitation of DNA immunization is that these responses are relatively weak and are often only transient in their nature. The use of immunologic adjuvants together with DNA vaccines is a promising way to enhance and to optimize DNA-derived immunity. Cytokines have been widely used to enhance the immune responses of DNA vaccines. In the present investigation, we studied the in vivo immunomodulation of HIV-1 Indian subtype C plasmid construct (pJWSK3, encoding envgp120 gene) by plasmid based murine IL-2/Ig construct. Subcloning of mIL-2/Ig gene from pVRCmIL-2/Ig construct into pJW4304 vector was done followed by its in vitro expression study on the COS-7 cell line. Co-immunization of the recombinant HIV-1 env-gp120 construct with the IL-2/Ig construct in the female Balb/c mice by the intramuscular route resulted in induction of significantly higher levels of both HIV-1-specific antibody response and cell mediated immune response than by DNA plasmid construct alone (p < 0.001 and p < 0.05, respectively). The induced HIV-1 specific murine IFN-gamma response was robust, broad based, and seen even at the end of 6 months after immunization. Taken together these results indicate that the strategy of using IL-2/Ig plasmid can be highly effective when used along with recombinant DNA constructs and serve as the potential tool for the development of more rationally designed vaccines against HIV-1. PMID- 16359232 TI - Recombinant adenovirus vector vaccine induces stronger cytotoxic T-cell responses than recombinant vaccinia virus vector, plasmid DNA, or a combination of these. AB - The efficiency of prime-boost vaccinations on the induction of T-cell responses to Sin Nombre virus nucleocapsid protein expressed by recombinant vaccinia virus, replication-deficient adenovirus, and plasmid DNA in mice was quantitated by the number of epitope-specific interferon-gamma-producing T cells and cytotoxic T lymphocyte activity induced. In prime-boost immunizations, all combinations that included the recombinant adenovirus induced a much higher number of epitope specific interferon-gamma-producing T cells than did other combinations. A single immunization of the recombinant adenovirus was able to induce similarly high levels of epitope-specific interferon-gamma-producing cells, despite the fact that the recombinant adenovirus produces less amount of the Sin Nombre virus nucleocapsid protein. PMID- 16359233 TI - Immunogenicity of the epitope of the foot-and-mouth disease virus fused with a hepatitis B core protein as expressed in transgenic tobacco. AB - A novel plant-based vaccine protecting against foot-and-mouth disease (FMD) was developed by inserting the VP21 epitope into the internal region of the hepatitis B virus core antigen gene (HBcAg). The specific sequence of the VP21 epitope is located within the VP1 capsid protein of the FMD virus (FMDV). It spans 21 amino acids located between positions 140 and 160 of the G-H loop. The fusion gene, HBCVP, was inserted into the plant binary vector pBI121 and then transformed into tobacco (Nicotiana tabacum) plants via Agrobacterium tumefaciens strain LBA 4404. The presence of HBCVP in the tobacco genome was confirmed by polymerase chain reaction (PCR); its transcription was verified by reverse transcription-PCR; and the recombinant protein expression was confirmed by Western blot analysis. The results of immunologic microscopic observation demonstrated that recombinant fusion protein HBCVP can form a virus-like particle (VLP) structure in transgenic tobacco leaves. Mice, immunized intraperitoneally with a soluble crude extract of transgenic tobacco leaves, were found to produce specific antibody responses to both HBcAg and FMDV VP1. A virus challenge demonstrated that the immunized mice were highly protected against virulent FMD. This work describes a new way to develop an FMD vaccine from plants that will aid the development of new vaccines using HBcAg fused to the conserved epitopes of other pathogenic antigens. PMID- 16359234 TI - Cross-clade protection induced by human immunodeficiency virus-1 DNA immunogens expressing consensus sequences of multiple genes and epitopes from subtypes A, B, C, and FGH. AB - The correlate of protection in human immunodeficiency virus (HIV) infection is not known, but preclinical and clinical studies support the involvement of both antibodies and cellular immunity. In addition, the existence of multiple HIV clades makes HIV vaccine design especially challenging. We have constructed a vaccine platform with an HIV-1 subtype B DNA immunogen expressing full length consensus sequences from HIV-1 rev, nef, tat, and gag with additional cellular epitope clusters from the env and pol regions. Furthermore, this platform has been extended to three additional plasmids expressing the same immunogens but originating from subtypes A or C consensus or FGH ancestral sequences. Immunogenicity in BALB/c mice, by gene gun or intramuscular delivery, revealed strong IFN-gamma production in response to in vitro re-stimulation with a H-2d restricted gag peptide (AMQMLKETI) or even stronger toward an env epitope (RGPGRAFVTI). Weak humoral immunity was detected. Gene gun immunization with a cocktail of all four plasmids induced pre-challenge cellular immunity in C57Bl6/A2.01 mice and subsequently a robust frequency of protection (11/12 animals) after experimental challenge with subtype A or B HIV-1/Murine Leukemia Virus (HIV-1/MuLV). The cross-clade protection observed in this challenge experiment demonstrates that these multigene/multiepitope HIV DNA immunogens are likely to be potent immunogens also against the HIV-infection of human beings. PMID- 16359235 TI - Timing of retroviral infection influences anamnestic immune response in vaccinated primates. AB - Using simian immunodeficiency virus (SIV) infection of rhesus macaques to model human immunodeficiency virus (HIV) infection of humans, we assessed whether broadly reactive vaccine-induced humoral immunity would remain broadly reactive after viral challenge, and whether there would be significant differences in anamnestic antibody responses if animals were challenged when predominately effector or memory lymphocyte populations were present. Animals immunized over a prolonged period and challenged 11 months after vaccination mounted more broadly reactive and stronger humoral immunity than those rapidly vaccinated and challenged 2 weeks after their final vaccinations. These data suggest that vaccination schedule and the timing of virus challenge should be considered when evaluating future candidate HIV vaccines. PMID- 16359236 TI - Mucosal and systemic adjuvant effects of cholera toxin and Cry1Ac protoxin on the specific antibody response to HIV-1 C4/V3 peptides are different and depend on the antigen co-administered. AB - Evidence from several sources support the assertion that cholera toxin (CT) is a potent immunogen and mucosal adjuvant; however there are also reports showing its lack of adjuvanticity against some antigens. Cry1Ac protoxin also exerts adjuvant effects in the antibody responses to proteins and polysaccharides but its adjuvanticity with regard to peptide vaccines had not been tested. To probe whether the adjuvant effects of these proteins varied depending on the antigen co administered, we evaluated antipeptide antibody responses in serum and mucosal samples (vaginal, intestinal, and pulmonary) of mice that were immunized by intranasal or intraperitoneal routes with one of two distinct hybrid C4/V3 HIV peptides, either alone or with CT or Cry1Ac. The tested HIV 1 peptides differed in two aminoacids, T1SP10MN(A) was modified at the SP10 region by the substitution of the isoleucines 12 and 14 for cysteines and was called modified (m)T1SP10MN(A). Our data indicate that the adjuvant effects of CT and Cry1Ac are different. In addition they vary depending on the antigen co-administered and the number of antigen doses, because after three doses moderate adjuvant effects of CT and Cry1Ac on anti-peptide serum and mucosal antibody responses were observed only against the mT1SP10MN(A). In contrast, to attain significant adjuvant effects against the T1SP10MN, four doses were required. Interestingly we found that modification of the HIV peptide increases its immunogenicity without altering the principal neutralizing determinant (SP10). PMID- 16359237 TI - DNA priming E and NS1 constructs--homologous proteins boosting immunization strategy to improve immune response against dengue in mice. AB - DNA priming-protein boosting is a strategy used to establish strong immunity to a specific pathogen by the use of two different antigens through sequential delivery systems. In this work, two recombinant plasmids were used, one encoding for the dengue virus E protein, which is know to induce neutralizing antibodies (pcDNA 3.1/E), and the other encoding for the Dengue virus nonstructural protein 1 (pcDNA 3.1/NS1), as a source of B- and T-cell epitopes possibly involved in protective immunity. We showed that immunization of BALB/c mice with three priming doses of both plasmids pcDNA 3.1/E and/or pcDNA 3.1/NS1 were able to induce antibody responses to E protein with a single plasmid; in contrast to the antibody response to NS1 protein we observed an additive effect in terms of antibody response. Moreover, using a prime-boost protocol in which both plasmid constructs were co-administrated followed by a boost of homologous GST-E and GST NS1 recombinant proteins, we observed an increased antibody response to NS1 and to E protein compared to animals vaccinated with the proteins or with dengue constructs alone. If neutralizing antibodies play an important role in dengue infection, antibodies generated with this regimen was also significantly better than the administration of the mix of proteins alone. These results suggest that NS1 and E proteins together could be considered in a design of subunit recombinant vaccines. PMID- 16359238 TI - Role of Indoleamine 2,3-Dioxygenase in Antiviral Activity of Interferon-gamma Against Vaccinia Virus. AB - Interferon-gamma (IFN-gamma) has antiviral activity against poxviruses as well as many other viruses, bacteria and a parasite, Toxoplasma gondii. Inducible nitric oxide synthase (NOS2) has been shown to mediate the antiviral activity of IFN gamma in both in vivo and in vitro experiments. In macrophages, inhibition of replication of poxviruses by IFN-gamma is NOS2-dependent. In this report we tested nonmacrophage cell lines and found that indoleamine 2,3-dioxygenase (IDO) also mediated the antiviral activity of IFN-gamma against vaccinia virus. L tryptophan, an inhibitor of IDO, completely blocked the antiviral activity of IFN gamma against vaccinia virus in 143B cells, an human osteosarcoma cell line, whereas N(G)-methyl-L-arginine, a NOS2 inhibitor, did not. IDO may account for the NOS2-independent antiviral mechanism induced by IFN-gamma. IFN-gamma may use different antiviral mechanisms in different cell types. PMID- 16359240 TI - High thymic volume is associated with viral replication and immunologic impairment only early after HAART interruption in chronic HIV infection. AB - One of the strategies that has been investigated to reduce antiretroviral treatment toxicity in patients infected with human immunodeficiency virus (HIV) is structured treatment interruption (STI). Our aim was to analyze early viral and immune dynamics after interruption of highly active antiretroviral therapy (HAART) and to determine whether thymic function-related markers play a role in preventing CD4 count decline caused by increased viral replication. This was a prospective study of an open cohort of 47 HIV-infected patients with a median 969 CD4 count and prolonged viral suppression. They were followed every 4 weeks though week 24. Thymic volume and TREC level were analyzed at baseline. Increased thymic volume was associated with higher plasma viral load and greater CD4 count decline early after interruption. Three virologic patterns were observed: rapid/high (RH), delayed/high (DH), and low/slow (LS) viral replication. RH correlated with higher thymic volume at baseline and with higher CD4 count decline at week 4. Patients with greater thymic volume was associated with an immune and virologic impairment only early after interruption, probably because of infection of the increased number of available target cells. As the long-term consequences of these observations are unknown, the safety of treatment interruption must be further studied. PMID- 16359239 TI - T cells expressing the Vgamma1 T-cell receptor enhance virus-neutralizing antibody response during coxsackievirus B3 infection of BALB/c mice: differences in male and female mice. AB - Coxsackievirus B3 infection causes severe cardiac inflammation in male but not female mice. CD3+ T cells and T cells expressing the Vgamma4 T cell receptor (TCR) predominate in the cardiac inflammatory cell infiltrate in infected male BALB/c mice. Infected females have mostly CD19+ (B lymphocyte) and Vgamma1+ cells. No significant differences in CD11b+ (monocyte) cells were observed between the sexes. Infected males showed a predominant CD4+Th1 (IFNgamma+) response, whereas females showed a predominant CD4+Th2 response. The importance of IFNgamma for myocarditis susceptibility and IL-4 for protection was confirmed using IFN-gamma-/- and IL-4-/- mice. Antibody depletion of Vgamma1+ cells augmented myocarditis susceptibility, whereas antibody depletion of Vgamma4+ cells was protective. Cardiac virus titers inversely correlated with virus neutralizing antibodies and showed that Vgamma1+ cells are important for virus neutralizing antibody response. IFNgamma affected the Vgamma4+ cell response in the heart, as IFNgamma-/- mice had few Vgamma4+ cells; but exogenous administration of recombinant IFNgamma to IFNgamma-/- mice restored myocarditis susceptibility, Th1 bias, and Vgamma4+ cell infiltration of the myocardium. These results demonstrate that two gammadelta+ T cell populations, Vgamma1+ and Vgamma4+, have different functions during myocarditis, in that Vgamma1+ cells promote humoral immunity and protection whereas Vgamma4+ cells are pathogenic. PMID- 16359242 TI - RSV 2005: Global impact, changing concepts, and new challenges. PMID- 16359245 TI - Microneedle array technology: the time is right but is the science ready? PMID- 16359246 TI - Is there a real alternative anticoagulant to heparin in continuous treatments? PMID- 16359247 TI - Tissue-engineering scaffolds: can we re-engineer mother nature? PMID- 16359248 TI - Needle-free injection technology. PMID- 16359250 TI - Scaffolds for liver tissue engineering. AB - This review focuses on the expanding role for biomaterials and polymer scaffolds in liver tissue engineering. Studies are subdivided into in vitro and in vivo approaches. The in vitro section of the review discusses the challenges specific to liver tissue engineering, and how the choice of scaffold and its structure influences the success of the regenerative medicine strategy. The in vivo section evaluates early attempts to stimulate liver repair with cell and growth factor therapies, their failings and how current approaches aim to solve these problems. PMID- 16359251 TI - Growth factor-delivery systems for tissue engineering: a materials perspective. AB - The transplantation of organs, their surgical reconstruction or implantation of synthetic devices that can perform the function of organs, are the currently available methods for treating loss of tissue/organs in humans. However, the limitations associated with these techniques have led to the development of tissue engineering. One of the primary goals of tissue engineering is to provide growth factor delivery systems that can induce desired cell responses both in vitro and in vivo, in order to cause accelerated tissue regeneration. To make growth factors a more therapeutically viable alternative for the treatment of chronic degenerative diseases, a wide range of natural and synthetic materials have been employed as vehicles for their controlled delivery. The choice of material and design of the carrier device influence the mode of immobilization of growth factors on the scaffolds and their local/systemic administration. From a tissue engineer's perspective, materials could be used for designing scaffolds as well as for delivering single or multiple growth factors. Therefore, this review discusses growth factor delivery systems, with particular reference to carrier based growth factor delivery systems with a focus on materials. PMID- 16359252 TI - Bone graft substitutes. AB - The current gold standard of bone grafts is the autograft since it possesses all the characteristics necessary for new bone growth, namely osteoconductivity, osteogenicity and osteoinductivity. However, the autograft has its limitations, including donor-site morbidity and supply limitations, hindering this as an option for bone repair. An extensive list of currently available alternatives to bone grafts is provided, along with a classification scheme that divides these bone graft substitutes into five groups depending on the primary material composition: allograft, cell, factor, ceramic and polymer. Of the bone graft substitutes listed, several are discussed in detail. Beyond the current state of the art, attention is paid to what lies ahead in the field of bone graft substitutes. Biodegradable composite structures and various new architectures are discussed, as are newly developed polymeric materials, with tissue engineering providing the platform for future directions within this discipline. PMID- 16359253 TI - Progress in the development of a corneal replacement: keratoprostheses and tissue engineered corneas. AB - Rapid progress has been made in the past 5 years in the development of corneal replacements. Traditionally they are divided into two categories, keratoprostheses and tissue-engineered corneal equivalents, as replacement tissues are increasingly in demand worldwide. There are currently several different keratoprosthesis models in clinical use around the world. The most popular and most widely publicized is the AlphaCor model, which has enjoyed significant clinical success. However, improvements remain to be made, and the aim of most of the current research is to better understand the interactions between a synthetic material and the surrounding biology on a more fundamental level. This improved understanding will no doubt lead to improvements in current models and to the development of new models in the near future. While tissue engineered corneal equivalents have been under investigation for considerably less time, there is growing evidence to suggest that a tissue-engineered corneal equivalent comprised of primarily natural materials will exist in the not too distant future. Research groups have reported strong in vitro and in vivo results. The strength of the collagen matrix and its ability to support cell infiltration have been the primary avenues of research. Various collagen crosslinking techniques have been used. Infiltration of three major cells of the cornea has been observed. Most importantly, the ability of these materials to support nerve ingrowth has been demonstrated. While challenges remain with both types of corneal replacements, the considerable progress in the recent past suggests that reliable implants for the treatment of a variety of corneal diseases will be available. This review will provide an overview of recent results, and will provide insight into the future of research on corneal replacements. PMID- 16359254 TI - Medical devices used for ureteroscopy for renal calculi. AB - Urinary stone disease is a problem as old as human civilization. In this article, a brief history of stone disease and the current status of urinary stone disease treatment will be discussed. This review will focus primarily on the newest technology and devices associated with flexible ureteropyeloscopy for the treatment of upper tract urinary stones. In addition to defining the cutting-edge technology that is newly available, this review will glimpse at what technologic advances are expected in the near future. Reference articles that do a good job of defining the current state of flexible ureteropyeloscopy technology and urinary stone disease treatment will be highlighted, along with a reiteration of the major points of the article. PMID- 16359255 TI - Therapeutic applications of lasers in urology: an update. AB - There has been renewed interest in the use of lasers for minimally invasive treatment of urologic diseases in recent years. The introduction of more compact, higher power, less expensive and more user-friendly solid-state lasers, such as the holmium:yttrium-aluminum-garnet (YAG), frequency-doubled neodymium:YAG and diode lasers has made the technology more attractive for clinical use. The availability of small, flexible, biocompatible, inexpensive and disposable silica optical fiber delivery systems for use in flexible endoscopes has also promoted the development of new laser procedures. The holmium:YAG laser is currently the workhorse laser in urology since it can be used for multiple soft- and hard tissue applications, including laser lithotripsy, benign prostate hyperplasia, bladder tumors and strictures. More recently, higher power potassium-titanyl phosphate lasers have been introduced and show promise for the treatment of benign prostatic hyperplasia. On the horizon, newer and more effective photosensitizing drugs are being tested for potential use in photodynamic therapy of bladder and prostate cancer. Additionally, new experimental lasers such as the erbium:YAG, Thulium and Thulium fiber lasers, may provide more precise incision of soft tissues, more efficient laser lithotripsy and more rapid prostate ablation. This review provides an update on the most important new clinical and experimental therapeutic applications of lasers in urology over the past 5 years. PMID- 16359256 TI - Laparoscopic splenectomy: present status and future perspective. AB - Laparoscopic splenectomy has become widely accepted as the approach of choice for the surgical treatment of benign and malignant hematologic diseases. Advances in technology have led to better outcomes for the procedure, and have allowed surgeons to apply the technique to disease processes that were at one time felt to be contraindications to laparoscopic splenectomy. However, challenges still remain. There is a steep learning curve associated with the procedure. The development of cost-effective laparoscopic simulators to target the skills required for laparoscopic splenectomy and other laparoscopic procedures is essential. The advent of devices which isolate and seal the large blood vessels that surround the spleen have reduced intra-operative bleeding and minimized conversions to open splenectomy. Improvements in optics and instrumentation, as well as robotic technology, will continue to define the frontier of minimally invasive surgery, and further facilitate the acceptance of laparoscopic splenectomy for the treatment of benign and malignant hematologic diseases. PMID- 16359257 TI - Laparoscopic duodenal switch for morbid obesity. AB - Laparoscopic duodenal switch gives a consistent excess weight loss of 70-80% with acceptable long-term nutritional complications. It is especially indicated for super-obese patients with a body mass index greater than 50 kg/m(2). A systematic review of the literature and results of open and laparoscopic duodenal switch is thoroughly presented. Also presented for the authors' surgical colleagues are some technical details concerning their preferred method. Meta-analysis now demonstrate a low mortality rate for the laparoscopic procedure close to 0.5%, and operative times close to 200 min. Laparoscopic duodenal switch is likely to increase in popularly for the treatment of morbid obesity, especially with the recent advent of laparoscopic sleeve gastrectomy for higher-risk patients. PMID- 16359258 TI - Diagnostic imaging for acute ischemic stroke management. AB - Although our clinical understanding remains our most important diagnostic tool, acute stroke therapy without neuroimaging is impossible. In most patients, only non-contrast computed tomography is used for diagnosis of acute stroke. However, findings based exclusively on clinical assessment and nonhemorrhagic computed tomography scans may no longer be appropriate for acute stroke treatment. From a pathophysiologic point of view, advanced computed tomography techniques and stroke magnetic resonance imaging provide much more information about the acute stroke patient as the basis of decision making in acute stroke treatment. Advanced computed tomography may provide information comparable with stroke magnetic resonance imaging, although a more detailed evaluation concerning these methods in clinical practice is required. This review gives the reader an integrated view on the current status of acute stroke imaging based on advanced computed tomography and multiparametric stroke magnetic resonance imaging protocols. These new imaging techniques allow for a far more individualized method of decision making according to the findings in each patient. This results in improved identification of patients with acute stroke syndromes, improved patient selection of those patients who are regarded suitable for thrombolysis, an extension of the rather strictly defined therapeutic time window for treatment, as well as a more sophisticated method of introduction of alternative therapies into clinical practice. PMID- 16359259 TI - Importance of rapid testing to combat the global threat of bird flu. PMID- 16359260 TI - What are the issues surrounding preimplantation genetic diagnosis for late-onset disorders? PMID- 16359262 TI - Putting science to work: novel diagnostic strategies in laboratory medicine. PMID- 16359263 TI - A universal array-based multiplexed test for cystic fibrosis carrier screening. AB - Cystic fibrosis is a multisystem autosomal recessive disorder with high carrier frequencies in caucasians and significant, but lower, carrier frequencies in other ethnicities. Based on technology that allows high detection of mutations in caucasians and significant detection in other ethnic groups, the American College of Medical Genetics (ACMG) and American College of Obstetricians and Gynecologists (ACOG) have recommended pan-ethnic cystic fibrosis carrier screening for all reproductive couples. This paper discusses carrier screening using the Tag-It multiplex mutation platform and the Cystic Fibrosis Mutation Detection Kit. The Tag-It cystic fibrosis assay is a multiplexed genotyping assay that detects a panel of 40 cystic fibrosis transmembrane conductance regulator mutations including the 23 mutations recommended by the ACMG and ACOG for population screening. A total of 16 additional mutations detected by the Tag-It cystic fibrosis assay may also be common. The assay method is described in detail, and its performance in a genetics reference laboratory performing high volume cystic fibrosis carrier screening is assessed. PMID- 16359264 TI - Miniaturization applied to analysis of nucleic acids in heterogeneous tissues. AB - Despite huge efforts in sample analysis, the measurement of marker nucleic acids within tissues remains largely nonquantitative. Gene analyses have benefited from sensitivity gains through in vitro gene amplification, including PCR. However, whilst these processes are intrinsically suited to highly reproducible, accurate and precise gene measurement, the term semiquantitative analysis is still commonly used, suggesting that other fundamental limitations preclude a generic quantitative basis to gene analysis. The most poorly defined aspect of gene analysis relates to the sample itself. The amount of cells and, particularly, cell subtype composition are rarely annotated before analysis; indeed, they are often extrapolated after analysis. To advance our understanding of pathogenesis, assay formats will benefit from resembling the dimensions of the cell, to assist in the analysis of cellular components of tissue complexes. This review is partly a perspective on how current miniaturization technologies, in association with molecular biology, microfluidics and surface chemistries, may evolve from the parts of a paradigm to enable the unambiguous quantitative analysis of complex biologic matter. PMID- 16359265 TI - DNA mutation detection and analysis using miniaturized microfluidic systems. AB - Identification of genetic sequence variations occurring on a population-wide scale is key to unraveling the complex interactions that are the underlying cause of many medical disorders and diseases. A critical need exists, however, for advanced technology to enable DNA mutation analysis to be performed with significantly higher throughput and at significantly lower cost than is currently attainable. Microfluidic systems offer an attractive platform to address these needs by combining the ability to perform rapid analysis with a simplified device format that can be inexpensively mass-produced. This paper will review recent progress toward developing these next-generation systems and discuss challenges associated with adapting these technologies for routine laboratory use. PMID- 16359266 TI - Advancement in characterization of genomic alterations for improved diagnosis, treatment and prognostics in cancer. AB - Most human cancers are characterized by genetic instabilities. These instabilities manifest themselves as a series of genetic alterations, including discrete mutations and chromosomal aberrations. With the human genome deciphered, high-throughput technologies are rapidly advancing the field to generate genome wide gene expression and mutation profiles that are highly correlative of biologic and disease phenotypes. While recent advancement in comprehensive genomic characterization presents an unprecedented opportunity for advancing the treatment of cancer, there are still many challenges that need to be overcome before we can fully utilize genomic markers and targets for cancer prediction, diagnostics, treatment and prognostics. This review describes recent advances in comprehensive genomic characterization at the DNA level, and considers some of the challenges that remain for defining the precise genomic portrait of tumors. Potential solutions that may help overcome these challenges are also offered. PMID- 16359267 TI - Quantification of chemokines by real-time reverse transcriptase PCR: applications in type 1 diabetes. AB - Type 1 diabetes is a T-cell mediated autoimmune disease, characterized by the destruction of insulin-producing pancreatic beta-cells. This review will discuss the role of chemokines in the recruitment of immune cells leading to the pathology of this disease. There will be a focus on the quantification of chemokines and chemokine receptors by the recently developed real-time reverse transcriptase PCR technique. Today, this technique is in widespread use for analysis of chemokines in cells, tissues and tissue biopsies. The minute amount of tissue needed for analysis, as well as the very high sensitivity of this method, make it the method of choice for analysis of chemokines, which are often expressed at very low levels in target tissues. However, validation and optimization of the technique is of crucial importance for obtaining reliable results. PMID- 16359268 TI - Molecular genetics in hypertrophic cardiomyopathy: towards individualized management of the disease. AB - Hypertrophic cardiomyopathy is a relatively common genetic disease, affecting one person per 500 in the general population, and is clinically defined by the presence of unexplained left ventricular hypertrophy. Although recognized as the most common cause of sudden death in the young (especially in athletes), the cardiac expression of the disease is highly variable with respect to age at onset, degree of symptoms and risk of cardiac death. As a consequence, therapeutic strategies are diverse and must be adapted to the specific features of an individual. Recently, the molecular bases of the disease have been unraveled with the identification of a large number of mutations in genes encoding sarcomeric proteins. This review focuses on the impact of the molecular data on the understanding of the disease, and considers the emerging issues regarding the impact of molecular testing on the management of patients (or relatives) in clinical practice. PMID- 16359269 TI - Molecular methods for the detection and characterization of Neisseria meningitidis. AB - Neisseria meningitidis remains a common global cause of morbidity and mortality. The laboratory confirmation of meningococcal disease is, therefore, very important for individual patient management and for public health management. Through surveillance schemes, it provides long-term epidemiologic data that can be used to inform vaccine policy. Traditional methods, such as latex agglutination and the enzyme-linked immunosorbent assay, are still used, but molecular methods are now also established. In this review, molecular methods for the laboratory confirmation and characterization of meningococci are described. PCR is an invaluable tool in modern biology and can be used to predict the group, type and subtype of meningococci. It is now also used in a fluorescence-based format for increased sensitivity and specificity. The method also provides the amplified DNA for other techniques, such as multilocus sequence typing. Other methods for the discrimination of meningococci have also played and continue to play an important part in epidemiology. For example, pulsed-field gel electrophoresis is highly discriminatory, whilst multilocus enzyme electrophoresis provided the basis for the description of global meningococcal clones and formed the foundation for multilocus sequence typing. Other less commonly used methods, such as matrix-assisted laser desorption/ionization time of-flight mass spectrometry and pyrosequencing, may increasingly find their way into microbiology reference laboratories. Nevertheless, nucleotide sequencing and laboratory automation have aided the introduction of many methods and provide data that are digitally based and, therefore, highly accurate and portable. PMID- 16359270 TI - Thin-film technology for direct visual detection of nucleic acid sequences: applications in clinical research. AB - Certain optical conditions permit the unaided eye to detect thickness changes on surfaces on the order of 20 A, which are of similar dimensions to monomolecular interactions between proteins or hybridization of complementary nucleic acid sequences. Such detection exploits specific interference of reflected white light, wherein thickness changes are perceived as surface color changes. This technology, termed thin-film detection, allows for the visualization of subattomole amounts of nucleic acid targets, even in complex clinical samples. Thin-film technology has been applied to a broad range of clinically relevant indications, including the detection of pathogenic bacterial and viral nucleic acid sequences and the discrimination of sequence variations in human genes causally related to susceptibility or severity of disease. PMID- 16359271 TI - A survey of methods for classification of gene expression data using evolutionary algorithms. AB - The rapid increase in the quantity of available biologic data over the last decade, brought about by the introduction of massively parallel methods for gene expression measurements, has highlighted the need for more efficient computational techniques for analysis. This paper reviews the use of evolutionary algorithms (EAs) in connection with classification based on gene expression data matrices. Brief introductions to data classification methods and EAs are given, followed by a survey of studies dealing with the application of evolutionary algorithms to various (cancer related) data sets. The general conclusion, based on the published results surveyed here, is that EAs may constitute an efficient method for optimal gene selection, and can also help in reducing the size (number of features used) of classifiers. In many cases, the classification accuracy obtained using EAs, often in conjunction with other methods, represents a significant improvement over results obtained without the use of EAs. However, long-term, independent clinical follow-up studies will be essential to validate prognostic markers identified by the use of EA-based methods. PMID- 16359272 TI - Antibody microarrays: the crucial impact of mass transport on assay kinetics and sensitivity. AB - Although they are superficially similar to DNA microarrays, immunoassay microarrays represent a daunting technological challenge owing to the much wider diversity of proteins. Yet, as the leading edge of bioscience migrates from genomics to proteomics, the complexity and enormous dynamic range of proteins in a cell necessitate an analytic tool with exceptional specificity and sensitivity. In theory, microspot immunoassays could fulfill this need. However, antibody microarrays have had limited success to date, and have often required a highly sensitive detection system and/or sophisticated immobilization approach to be of any use for the profiling of complex specimens. There is a solid body of work on the theory of microspot reaction kinetics, yet much of the published experimental work on protein microarray development pays insufficient attention to the kinetic aspects of this interaction. This review explains that one of the main limitations for the sensitivity of current generation microspot immunoassays is the strong dependence of antibody microspot kinetics upon mass flux to the spot. This not only involves migration of analyte in solution, but also across the surface of the solid phase. Understanding of this effect will be discussed, along with several related effects and their significance to improving existing microarray designs. It is concluded that current efforts may be too focused on areas that cannot improve performance significantly, and that other critical areas of design should receive more attention. Finally, the review addresses the question of whether ambient analyte immunoassay is truly a separate category of microspot assay, with the conclusion that this may be a flawed concept. PMID- 16359273 TI - Marshall M. Parks, M.D. 1918-2005. PMID- 16359274 TI - Marshall M. Parks, M.D. memorial issue: remembrances. PMID- 16359275 TI - Dedication of this issue to the late great Marshall M. Parks. PMID- 16359276 TI - A small-incision muscle hook for the Parks cul-de- sac approach for strabismus surgery. AB - BACKGROUND: One of Dr. Marshall Parks' most valuable contributions to strabology was his cul-de-sac ("fornix") approach to strabismus surgery. This approach provides better comfort, faster healing and less conjunctival scarring than previous approaches. Tearing of the conjunctiva can occur, however, especially in older patients with thin and friable conjunctival tissues. METHODS: To facilitate the cul-de-sc approach through a small incision, especially in older patients, I designed a new "small-incision" muscle hook. RESULTS: A new sharp second bend combined with sharpening of the bend in the heel (creating literally an "S" bend) in the Jameson style eye muscle hook enables burying of the knee or heel of the hook in the wound, allowing reflection of the conjunctiva over the ball or toe of the hook without tearing. I have used and treasured this hook for over six years, routinely using the cul-de-sac approach to strabismus surgery in both children and adults. CONCLUSIONS: This new small- incision muscle hook complements the cul de-sac approach to strabismus surgery by allowing the use of a smaller incision while significantly decreasing the risk of conjunctival tearing. PMID- 16359277 TI - Video vision development assessment in diagnosis and documentation of microtropia. AB - BACKGROUND: Microtropia is under diagnosed in treated esotropia and in primary microtropia cases, where patients are young and uncooperative. METHOD: Video Vision Development Assessment (VVDA) testing, which we have developed and previously described, captures multiple video frames images (30 per second) of the Breuckner red reflection (from the ocular fundus, a test for strabismus and ocular media abnormalities) combined with eccentric photorefraction. This method allows the highly critical discrimination of minimally off axis fixation (abnormal) to be differentiated from true on axis fixation (normal foveation) by the brightness difference in the images. We have examined with this method, VVDA, 533 consecutive strabismus patients ages six months to six years. RESULTS: 47% of esotropes and 23.8% of exotropes had microtropia. CONCLUSION: VVDA is helpful in documenting microtropia in young uncooperative patients where the diagnosis may otherwise be missed, even when suspected to be present, due to lack of subjective test cooperation. Multiple video frames of the same patient (VVDA) allow dynamic detection of true fixation (normal foveation) versus slightly off axis fixation (abnormal, consistent with microtropia). VVDA is therefore superior to single photorefraction imaging in the diagnosis of microtropia. PMID- 16359278 TI - Sherrington innervational surgery in the treatment of chronic sixth nerve paresis. AB - INTRODUCTION AND PURPOSE: To describe a new operation to treat unilateral chronic sixth nerve paresis based on Sherrington's innervational law. A recession of the medial rectus (MR) in the good eye, yoke to the paretic lateral rectus (LR), will have the reciprocal innervational effect of relaxing the contracture of the contralateral MR and by doing so will enhance the effect of a weakening procedure performed on this muscle. The goal of this study was to eliminate diplopia in primary position by improving the function of the paretic LR and reducing the contracture of its antagonist MR. METHODS: The records of 14 consecutive patients with unilateral chronic sixth nerve paresis so treated were reviewed. Nine had bilateral medial rectus muscle retroplacement and postop' adjustable sutures. A non-adjustable resection of the paretic lateral rectus muscle was added to the other five. Average time from onset to surgery was 60 months (minimum 9 months). Average post-surgical followup was 22 months. RESULTS: The function of the paretic LR and the contracture of the ipsilateral MR were improved in all 14 cases. Patients with bilateral medial rectus recessions and postop' adjustable sutures had an average correction of 32 prism diopters in primary position. Patients with the added resection of the paretic LR had an average correction in primary position of 46 prism diopters. Two of the 14 patients failed our goals; one had residual diplopia in primary position and the other one had diplopia within 30 degrees on gaze to one side; for an 86% success rate. CONCLUSIONS: The 86% success rate in this study (ultimately we also achieved a 100% satisfaction rate) indicates that innervational surgery in the form of a recession of the MR in the good eye added to that of the MR in the involved eye in patients with unilateral chronic sixth nerve paresis is a safe and effective surgical procedure. PMID- 16359279 TI - Visualization of actin cytoskeletal dynamics during the cell cycle in tobacco (Nicotiana tabacum L. cv Bright Yellow) cells. AB - BACKGROUND INFORMATION: The actin cytoskeleton forms distinct actin arrays which fulfil their functions during cell cycle progression. Reorganization of the actin cytoskeleton occurs during transition from one actin array to another. Although actin arrays have been well described during cell cycle progression, the dynamic organization of the actin cytoskeleton during actin array transition remains to be dissected. RESULTS: In the present study, a GFP (green fluorescent protein) mTalin (mouse talin) fusion gene was introduced into suspension-cultured tobacco BY-2 (Nicotiana tabacum L. cv Bright Yellow) cells by a calli-cocultivation transformation method to visualize the reorganization of the actin cytoskeleton in vivo during the progression of the cell cycle. Typical actin structures were indicated by GFP-mTalin, such as the pre-prophase actin band, mitotic spindle actin filament cage and phragmoplast actin arrays. In addition, dynamic organization of actin filaments was observed during the progression of the cell from metaphase to anaphase. In late metaphase, spindle actin filaments gradually shrank to the equatorial plane along both the long and short axes. Soon after the separation of sister chromosomes, actin filaments aligned in parallel at the cell division plane, forming a cylinder-like structure. During the formation of the cell plate, one cylinder-like structure changed into two cylinder-like structures: the typical actin arrays of the phragmoplast. However, the two actin arrays remained overlapping at the margin of the centrally growing cell plate, forming an actin wreath. When the cell plate matured further, an actin filament network attached to the cell plate was formed. CONCLUSIONS: Our results clearly describe the dynamic organization of the actin cytoskeleton during mitosis and cytokinesis of a plant cell. This demonstrates that GFP-mTalin-transformed tobacco BY-2 cells are a valuable tool to study actin cytoskeleton functions in the plant cell cycle. PMID- 16359280 TI - The impact of catecholamines on defibrillation threshold in patients with implanted cardioverter defibrillators. AB - OBJECTIVES: To determine the effect of physiologic catecholamine concentrations on the defibrillation threshold (DFT) in patients with implanted cardioverter defibrillators. BACKGROUND: DFT is the minimum energy delivered by an implanted cardioverter defibrillator that successfully converts ventricular fibrillation. DFT testing is performed under conscious sedation. Since activities of daily living enhance sympathetic tone substantially over these nadir levels, it is important to explore the impact of catecholamines on DFT. METHODS: In this double blind study, we determined DFT by the step-down method. Patients (n = 50) were stratified by beta-blocker use and then randomized to a 7-minute infusion of epinephrine, norepinephrine, or placebo. After study infusion, DFT testing was repeated. Changes in DFT with different study medications were compared. Subgroup analyses of the effects of catecholamines on DFT, based on beta-blocker use, were also performed. RESULTS: Norepinephrine reduced DFT from baseline measurements by 22.6% (P = 0.008). Neither epinephrine nor placebo impacted DFT (P = 0.999, P = 0.317, respectively). In the subgroup analyses, DFT was reduced with norepinephrine regardless of beta-blocker use, while epinephrine reduced DFT among those receiving beta-blockers. No change in DFT was observed in either of the placebo subgroups. CONCLUSIONS: Elevation of plasma norepinephrine concentrations reduces the DFT, while elevations in epinephrine had no effect. Norepinephrine seems to reduce DFT regardless of beta-blocker therapy but epinephrine's effects are beta-blocker dependent. PMID- 16359281 TI - Long-term outcomes of a telephone intervention after an ICD. AB - BACKGROUND: The purpose of this study was to determine the long-term benefits of participating in a structured, 8-week educational telephone intervention delivered by expert cardiovascular nurses post-ICD. The intervention was aimed to (1) increase physical functioning, (2) increase psychological adjustment, (3) improve self-efficacy in managing the challenges of ICD recovery, and (4) lower levels of health care utilization over usual care in the first 12 months post ICD. This article reports on the 6- and 12-month outcomes of the nursing intervention trial. METHODS AND RESULTS: A two-group (N = 168) randomized control group design was used to evaluate intervention efficacy with persons receiving an ICD for the secondary prevention of sudden cardiac arrest. Measures were obtained at baseline, 6 and 12 months post hospitalization. Outcomes included (1) physical functioning (Patient Concerns Assessment [PCA], Short Form Health Survey [SF-12], ICD shocks), (2) psychological adjustment (State-Trait Anxiety Inventory [STAI], Centers for Epidemiologic Studies-Depression [CES-D], fear of dying), (3) self efficacy (Sudden Cardiac Arrest-Self-Efficacy [SCA-SE], Sudden Cardiac Arrest Behavior [SCA-B], Sudden Cardiac Arrest-Knowledge [SCA-K]), and (4) health care utilization (emergency room [ER] visits, outpatient visits, hospitalizations). Using repeated measures ANOVA, the 6- and 12-month benefits of the intervention over usual care were in reductions in physical concerns (P = 0.006), anxiety (P = 0.04), and fear of dying (P = 0.01), with enhanced self-confidence (P = 0.04) and knowledge (P = 0.001) to manage ICD recovery. There were no statistically significant differences between the groups on total outpatient visits, hospitalizations, or ER visits over 12 months. CONCLUSION: A structured 8-week post-hospital telephone nursing intervention after an ICD had sustained 12-month improvements on patient concerns, anxiety, fear of dying, self-efficacy, and knowledge. Results may not apply to individuals with congestive heart failure who receive an ICD for primary prevention of sudden cardiac arrest. PMID- 16359282 TI - Adaptive cardiac resynchronization therapy device: a simulation report. AB - We report the results of a simulation of an adaptive cardiac resynchronization therapy (CRT) device performing biventricular pacing in which the atrioventricular (AV) delay and interventricular (VV) interval parameters are changed dynamically in response to data provided by the simulated IEGMs and simulated hemodynamic sensors. A learning module, an artificial neural network, performs the adaptive part of the algorithm supervised by an algorithmic deterministic module, internally or externally from the implanted CRT or CRT-D. The simulated cardiac output obtained with the adaptive CRT device is considerably higher (30%) especially with higher heart rates than in the nonadaptive CRT mode and is likely to be translated into improvement in quality of life of patients with congestive heart failure. PMID- 16359283 TI - Biochemical evidence of cardiac damage following transvenous implantation of a permanent antibradycardia pacemaker lead. AB - OBJECTIVES: We tested the hypothesis that transvenous permanent pacemaker lead implantation causes clinically detectable myocardial damage. BACKGROUND: Histological evidence of myocardial damage has been reported after antibradycardia pacemaker lead implantation. METHODS: We studied 30 patients undergoing implantation of a full antibradycardia pacemaker system (pulse generator plus leads) and 10 patients in whom only a generator was implanted. Blood samples for cardiac troponin-I (CTNI), CK-MB mass, and myoglobin measurement were drawn at baseline, at the end of the procedure, and at 2, 6, 12, 24, 48, and 72 hours thereafter. RESULTS: Abnormal CTNI levels were noted only in 24 of the 30 patients undergoing a full system implantation. CTNI levels were already abnormal at the end of the procedure in 16 and became so in all 24 during the next 6 hours. Peak levels were reached within 6 hours in 21 patients and were compatible with "minimal" necrosis (CTNI < 1.5 pg/mL) in 20. Maximum ventricular lead diameter and number of implanted leads were independent predictors of peak CTNI levels. CK-MB mass also increased after the procedure, but exceeded the normal range in only 10 patients. Myoglobin levels increased significantly both in patients undergoing a complete system implantation and in those where only a pulse generator was implanted. CONCLUSIONS: Transvenous insertion of endocardial leads for permanent pacing is accompanied in most patients by "minimal" myocardial damage. In this setting CTNI level kinetics are fast, characterized by early elevation and peak. PMID- 16359284 TI - Prolonged paced QRS duration as a predictor for congestive heart failure in patients with right ventricular apical pacing. AB - BACKGROUND: The recent studies showed that right ventricular (RV) pacing was associated with worsening of heart failure. The aim of this study is to clarify the clinical significance of paced QRS duration during RV pacing to predict congestive heart failure (CHF) patients. METHODS AND RESULTS: This study enrolled in 92 patients with atrioventricular block who underwent initial pacemaker implantation. The paced QRS duration was automatically obtained by electrocardiography immediately after pacemaker implantation and then by routine attendance at a pacemaker clinic every 3 months. The paced QRS duration was positively correlated with left ventricular end-diastolic dimension (P < 0.05) and left ventricular end-systolic dimension (P < 0.05), and tended to negatively correlate with left ventricular ejection fraction (P = 0.0507). The paced QRS duration immediately after pacemaker implantation was 170.4 +/- 18.9 ms. During a mean follow-up period of 53 +/- 16 months, 16 patients developed CHF. We selected as a cut-off value the nearest whole number (190 ms) that was one standard deviation greater than the mean, and divided into two groups according to baseline paced QRS duration. Patients with a paced QRS duration of <190 ms comprised group A (n = 77, nine of which developed CHF) and the remainder comprised group B (n = 15, seven of which developed CHF). Prolonged paced QRS duration (> or =190 ms) was associated with a significant increase in the overall morbidity of CHF (P < 0.05). Additionally, paced QRS duration significantly prolonged during the follow-up period among group A patients with CHF (P < 0.05), but did not change among patients without CHF. CONCLUSION: We concluded that paced QRS duration can be a useful indicator of impaired left ventricular function in patients with RV pacing. Even in patients whose paced QRS duration is relatively shorter, progressive prolongation of paced QRS duration can predict the development of CHF. PMID- 16359285 TI - Abnormal heart rate turbulence predicts the initiation of ventricular arrhythmias. AB - BACKGROUND: Abnormal heart rate turbulence (HRT) reflects autonomic derangements predicting all-cause mortality, yet has not been shown to predict ventricular arrhythmias in at-risk patients. We hypothesized that HRT at programmed ventricular stimulation (PVS) would predict arrhythmia initiation in patients with left ventricular dysfunction. METHODS: We studied 27 patients with coronary disease, left ventricular ejection fraction (LVEF) 26.7 +/- 9.1%, and plasma B type natriuretic peptide (BNP) 461 +/- 561 pg/mL. Prior to arrhythmia induction at PVS, we measured sinus cycles after spontaneous or paced premature ventricular contractions (PVCs) for turbulence onset (TO; % cycle length change following PVC) and slope (TS; greatest slope of return to baseline cycle). T-wave alternans (TWA) was also measured during atrial pacing. RESULTS: At PVS, abnormal TO (> or =0%) predicted inducible ventricular tachycardia (VT; n = 10 patients; P < 0.05). TO was greater in inducible than in noninducible patients (2.3 +/- 3.1% vs -0.02 +/- 2.8%, P < 0.05) and correlated with LVEF (P < 0.05) but not with BNP. TS did not differ between groups. Conversely, ambulatory HRT differed significantly from HRT at PVS (TO -0.55 +/- 1.08% vs 0.85 +/- 3.02%, P < 0.05; TS 2.63 +/- 2.09 ms/RR vs 8.70 +/- 6.56 ms/RR, P < 0.01), and did not predict inducible VT but trended (P = 0.05) to predict sustained VT on 739 +/- 179 days follow-up. TWA predicted inducible (P < 0.05) and spontaneous (P = 0.0001) VT but did not co migrate with HRT. CONCLUSIONS: Abnormal HRT measured at PVS predicted the induction of sustained ventricular arrhythmias in patients with ischemic cardiomyopathy. However, HRT at PVS did not correlate with ambulatory HRT, nor with TWA, both of which predicted spontaneous ventricular arrhythmias. Thus, HRT may reflect the influence of autonomic milieu on arrhythmic susceptibility and is likely complementary to traditional arrhythmic indices. PMID- 16359286 TI - Rhythm abnormalities in children with isolated ventricular noncompaction. AB - BACKGROUND: Isolated ventricular noncompaction (IVNC) is a serious cardiomyopathy with a generally poor prognosis. It is characterized by the presence of prominent ventricular myocardial trabeculations and deep intertrabecular recesses, in the absence of other structural heart defects. This cardiomyopathy is usually associated with ventricular dysfunction, thromboembolic events, and rhythm problems. METHODS AND RESULTS: This article describes 11 children who have rhythm abnormalities associated with IVNC. On admission, eight children had complaints including palpitation, syncope, heart failure, and bradycardia. Ventricular arrhythmias were diagnosed in four children, sinus node and atrioventricular conductance disturbances in six children, and Wolff-Parkinson-White syndrome and associated tachycardia in one child. Three children with ventricular arrhythmias received an automatic implantable cardioverter defibrillator (ICD). Three patients died of cardiac problems during the follow-up period. CONCLUSION: Since many of rhythm abnormalities, including life-threatening ventricular arrhythmias, may be seen in patients with IVNC, children with IVNC should be screened for arrhythmias. An ICD may be the best treatment for some of these patients. PMID- 16359287 TI - Long-term mortality in patients with pauses in ventricular electrical activity. AB - BACKGROUND: The long-term significance of ventricular pauses of > or =3.0 seconds observed on Holter monitor is unclear, as previously conducted retrospective studies have been poorly controlled. We compared the prognosis of patients with pauses > or =3.0 seconds on Holter monitor with a well-matched control group without such pauses. METHODS: Scanning the Holter database at Ochsner Clinic (n = 11,730; January 1998 to June 2003) for pauses > or =3.0 seconds identified 70 patients (pause group). Of those, 29 (37.1%) received a permanent pacemaker (PPM group) and 41 (62.9%) did not (No-PPM group). For each No-PPM patient, two patients without pauses (<2.0 seconds) exactly matched for age, sex, ejection fraction (EF), rhythm, and duration of follow-up were randomly chosen from the Holter database (control group, n = 82) and survival of the two groups was compared. RESULTS: Mean age was 72.5 +/- 15.0 years, mean EF was 52.2 +/- 12.7%, and 68.3% were men. Mean follow-up was 2.2 years (0.5-4.5 years). There was no difference in survival between the No-PPM and the control groups (82.9% vs 84.1%, P = NS). Compared with the PPM group, pauses in the No-PPM group were more commonly asymptomatic, nocturnal, and due to sinus pauses or atrial fibrillation (AF) with slow ventricular response. CONCLUSIONS: Pauses in ventricular electrical activity > or =3 seconds on Holter monitor due to sinus pauses or AF with slow ventricular response are not predictive of heightened mortality. PMID- 16359288 TI - Heart rate variability is a useful parameter for evaluation of anticholinergic effect associated with inducibility of atrial fibrillation. AB - BACKGROUND: Disopyramide is thought to have an advantageous effect for atrial fibrillation (AF) associated with vagal activity because of its anticholinergic effect. METHOD: We used a canine vagal nerve stimulation (VNS) model. The monophasic action potential (MAP) duration at 90% repolarization (MAP(90)), the intraatrial conduction time, the induciblity of AF by electrical stimulation, and the amplitude of high-frequency component (HF-amp) of the heart rate variability (HRV) were evaluated before and after the administration of disopyramide (1 mg/kg) (group D, n = 8) or pilsicainide (1 mg/kg) (group P, n = 5). RESULTS: In group D, HF-amp decreased in the baseline condition from 1.1 +/- 0.6 to 0.6 +/- 0.4 ms and the degree of VNS-induced augmentation of HF-amp was attenuated from +492% to +127%. VNS-induced shortening of MAP(90) was also attenuated in the right atrium (from -30 +/- 15% to -10 +/- 6%) and in the left atrium (from -15 +/ 9% to -6 +/- 6%). In group P, little effect was shown in these parameters. The vagotonic AF became noninducible in all eight experiments in group D, while in only one of five in group P. CONCLUSION: The beneficial effect of disopyramide for vagotonic AF is based on the decrease of basal vagal tone and attenuation of the effect of vagal stimulation in the atrial myocardium. HRV is a useful parameter for evaluation of the effect of antiarrhythmic drugs on the autonomic nerve system, and the evaluation of variability may be useful for testing drug efficacy for arrhythmias. PMID- 16359289 TI - Correlation of electrotonic monophasic action potential shortening with short term memory in human atrium. AB - To determine the presence of memory in human atria, we recorded monophasic action potential (MAP) at the high right atrium (HRA) in 21 patients. After reaching a steady state at 600 ms, HRA pacing was switched to the coronary sinus (CS) pacing to alter the activation sequence. After 20 minutes of CS pacing, pacing was continued at HRA to record the memory effect of CS pacing. Atrial memory was defined as the change in HRA MAP duration (MAPd) after 20 minutes of altered activation sequence. Baseline MAPd was 229 +/- 31 ms, which was shortened to 226 +/- 24 ms immediately after CS pacing. After 20 minutes of CS pacing, HRA MAPd during HRA pacing was 220 +/- 28 ms, which was significantly shorter than the baseline MAPd (P = 0.003). The degree of atrial memory was associated with the degree of initial electrotonic MAPd changes caused by the altered activation sequence. These results suggest that memory phenomenon exists in human atria, and it can be expressed as a change in MAPd. PMID- 16359290 TI - Polymorphic ventricular tachycardia with a normal QT interval following azithromycin. PMID- 16359291 TI - Apropos "tachycardia begets tachycardia". AB - It has been shown that a tachycardia can facilitate the induction of the same ("atrial fibrillation begets atrial fibrillation") or a different tachycardia. This may also apply for pacemaker-mediated tachycardia as the present case documents. PMID- 16359292 TI - Cardiovascular abnormalities and arrhythmias in patients with Ondine's curse (congenital central hypoventilation) syndrome. AB - Patients with congenital central hypoventilation syndrome (CCHS) (Ondine's curse syndrome) have impaired autonomic control of ventilation with intact voluntary control of respiration. Autonomic dysfunction and cardiac abnormalities are common in CCHS. Bradyarrhythmias are life-threatening and often require pacemaker insertion. We presented a case of a patient with CCHS suffering from long sinus pauses requiring cardiac pacemaker insertion. Patients with CCHS are at risk for pulmonary hypertension and cor pulmonale secondary to chronic hypoxia. Diaphragmatic pacing has been beneficial in some patients with CCHS. In this article, we review concomitant cardiac abnormalities and the occurrence of bradyarrhythmias in patients with CCHS. PMID- 16359293 TI - "You have to make a contribution". PMID- 16359294 TI - The history of AV nodal reentry. AB - Though patients with AV nodal reentry are now routinely cured by catheter ablation, the basic mechanism of this disorder is still under debate. The putative mechanism of AV node reentry was first discovered by the elegant work of Gordon Moe. He demonstrated the existence of dual pathways and echo beats in rabbits. Building on these seminal observations, the mechanism of AVNRT has burgeoned to include the possibility of left atrial input into the node. The first curative nonpharmacologic procedures involved surgical dissection around the AV node and the procedure was rapidly supplanted by catheter ablation procedures. The initial ablative procedure targeted the fast pathway, but later observations showed that ablation of the slow pathway was more effective and safer. Cure of AV nodal reentry which is the most common cause of paroxysmal supraventricular tachycardia became possible through the cooperative efforts of anatomists, physiologists, surgeons, and clinical electrophysiologists. PMID- 16359295 TI - Absence of device-device interaction (DDI) in a patient with cardiac and diaphragmatic pacemakers for congenital central hypoventilation syndrome. AB - Autonomic control of ventilation is impaired in patients with Ondine's curse or congenital central hypoventilation syndrome (CCHS), but voluntary control remains intact. Bradyarrhythmias can be life threatening. In a patient with CCHS and long sinus pause requiring cardiac pacemaker insertion, a diaphragmatic pacemaker inserted in early childhood caused diaphragmatic pacer spikes observed during the interrogation of the cardiac pacemaker. Diaphragmatic pacing did not interfere with the cardiac pacemaker function. PMID- 16359296 TI - Cardiac resynchronization therapy using a VDD lead. AB - In heart failure patients with normal sinus node function, cardiac resynchronization therapy can be achieved with only two leads, one VDD type, and one left ventricular. This reduces the number of venous punctures, implanted leads, and possibly operation and fluoroscopic times and complication rates. We present two cases and discuss the advantages and limits of such a procedure. PMID- 16359297 TI - Balloon occlusion technique to cannulate angulated and tortuous coronary sinus branches in cardiac resynchronization therapy. AB - We present two cases that demonstrate a new technique to cannulate angulated and tortuous coronary sinus branches during left ventricular lead placement for cardiac resynchronization therapy. The technique uses an occlusive pulmonary artery balloon just beyond the takeoff of the coronary sinus branch to assist in the cannulation of the branch. PMID- 16359298 TI - Persistent left superior vena cava-inferior vena caval communication complicating implantation of an implantable cardioverter defibrillator. AB - A persistent left superior vena cava has been observed in 0.3% of the general population on autopsy. Its presence can complicate left-sided device implantation. Commonly, a LSVC connects to the right atrium via the coronary sinus. A LSVC-accessory hemiazygous-hemiazygous-inferior vena caval communication has not been described previously. The presence of such a congenital venous anomaly will prohibit a left-sided device implant. PMID- 16359299 TI - Unsuccessful AV nodal ablation in atrial fibrillation: an alternative method to achieve complete heart block. AB - We report the case of an elderly patient who presented with poorly tolerated episodes of atrial arrhythmia refractory to medical treatment. AV node ablation was identified as the only alternative expected to be efficacious for symptom relief. However, this usually simple intervention failed. The goal of creating a complete AV block was finally achieved through ablation of the anterior fascicle of the His bundle, which represented the only pathway for residual conduction in this patient. PMID- 16359300 TI - Successful radiofrequency ablation of ventricular premature contractions within the coronary sinus. AB - This case report describes frequent ventricular premature contractions (VPCs) originating from an epicardial site of the basal posteroseptal ventricle. Detailed pace-mapping using a high output pacing stimulator was utilized to successfully ablate VPCs within the coronary sinus. PMID- 16359303 TI - Clinical, immunological and histopathological findings in a subpopulation of dogs with pododermatitis. AB - Clinical, immunological and histopathological findings in 20 adult dogs of varying breeds with chronic (>or=6 months) inflammation confined to the pedal skin were compared over a 2-year period with those of a group of age-matched controls (n=20). All affected dogs were pruritic but systemically well. Lesions were present on all four feet in 18/20 cases. Affected feet were characteristically erythematous, swollen, painful and alopecic. Sinus tracts were evident in 4/20 dogs. Despite a methodical series of diagnostic tests, no underlying cause was identified. None of the dogs responded to antimicrobial therapy administered for 8 weeks, none had evidence of ectoparasitism and none satisfied the criteria for atopic dermatitis. There was no response to a dietary trial using a novel protein source. The condition was characterized histopathologically by epidermal hyperplasia, hyperkeratosis, spongiosis, dermal oedema and perivascular aggregates of lymphocytes and plasma cells. Clinical signs did not correlate with histopathological findings. Affected dogs had significantly elevated serum IgG and IgM concentrations. The results of lymphocyte proliferation assays and phenotypic studies to determine the relative percentage of CD3+, CD4+, CD8+ and CD21+ lymphocyte subsets, and the ratio of CD4+/CD8+ cells were not significantly different between groups. No age, sex or seasonal predilections were noted. All dogs subsequently responded to immunosuppressive doses of prednisolone or cyclosporin. The term immunomodulatory responsive lymphocytic-plasmacytic pododermatitis is proposed to denote what may be a previously unrecognized condition in some dogs with pododermatitis of undetermined aetiology. PMID- 16359304 TI - Alterations of epidermal proliferation and cytokeratin expression in skin biopsies from heavy draught horses with chronic pastern dermatitis. AB - We report the historical, clinical and histopathological characteristics of skin lesions in biopsies from 37 heavy draught horses with chronic pastern dermatitis. The skin lesions were divided into four macroscopic groups: scaling (group I, n=5), hyperkeratotic and hyperplastic plaque-like lesions (group II, n=14), nodular skin masses (group III, n=16) and verrucous skin lesions (group IV, n=2). The principal histological findings were hyperkeratosis and epidermal hyperplasia. There was a gradual increase in epidermal hyperplasia from groups I to IV, suggesting that the lesions represent different stages of disease. In all cases, there was perivascular dermatitis dominated by T lymphocytes with an increase in MHC class II-positive dendritic-like cells. Immunohistochemical labelling for cytokeratins CK5/6(4), CK10 and CK14 indicated a change in their expression pattern. This correlated with the degree of epidermal hyperplasia, indicating abnormal differentiation of keratinocytes. There was a statistically significant correlation between the severity of skin lesions and several other factors including increasing age, increasing cannon circumference, prominence of anatomical structures such as fetlock tufts of hairs, ergots and chestnuts, and bulges in the fetlock region. PMID- 16359305 TI - Adherence of Staphylococcus intermedius to corneocytes of healthy and atopic dogs: effect of pyoderma, pruritus score, treatment and gender. AB - Staphylococcal pyoderma occurs commonly in atopic dogs. Some studies have suggested that adherence of staphylococci to corneocytes of atopic dogs and humans is higher than to corneocytes of healthy individuals. This hypothesis and possible differences resulting from the presence or absence of pyoderma, the severity of pruritus or the effect of treatment or gender, were studied. Adherent bacteria (Staphylococcus intermedius) were quantified by computerized image analysis on corneocytes collected from healthy or atopic dogs using double-sided adhesive tape. The adherence of S. intermedius to the corneocytes of atopic dogs was significantly greater than to those of healthy dogs (P=0.005). Furthermore, adherence was significantly greater in dogs with high levels of pruritus compared to those with low scores. No significant differences were found between atopic dogs with no history of pyoderma, atopic dogs with a history of pyoderma and atopic dogs with pyoderma at the time of sampling (P=0.068), suggesting that factors other than adherence are necessary for clinical pyoderma to develop. Treatment did not generally influence the adherence of S. intermedius to corneocytes of atopic dogs and there was no gender difference in adherence in either healthy or atopic dogs. PMID- 16359306 TI - Linkage of dermatomyositis in the Shetland Sheepdog to chromosome 35. AB - Dermatomyositis is an inflammatory disease of the skin and muscle and is most commonly found in the Shetland sheepdog. Both the clinical presentation and the age of onset of dermatomyositis vary widely, and the inability to diagnose dermatomyositis before clinical symptoms ensue has made control of the disease difficult. Identification of a genetic marker that cosegregates with dermatomyositis would facilitate the development of a DNA-based test for the early detection of affected dogs. We report the use of linkage disequilibrium (LD) mapping to identify linkage to phenotypic dermatomyositis in the Shetland sheepdog. One marker, microsatellite marker FH3570 on canine chromosome 35, had evidence of LD (P=0.00002). Further studies are necessary to narrow the region harbouring the dermatomyositis locus, identify candidate genes and determine mode of inheritance. PMID- 16359307 TI - Pustular calicivirus dermatitis on the abdomen of two cats following routine ovariectomy. AB - An unusual form of calicivirus dermatitis is described in two cats. Two fully vaccinated cats were re-admitted for anorexia and depression following routine ovariectomy. Signs of upper respiratory disease were not present. One cat subsequently showed painful necrosis of the incision wound, the other one developed dyspnoea with pleural effusion and discrete tongue ulcers. Intact pustular lesions confined to the surgically prepared abdomen appeared in both cats, respectively, on days 11 and 9. The histopathological diagnosis was panepidermal pustulosis and necrotizing dermatitis. Positive immunohistochemical staining consistent with feline calicivirus antigen was detected in epithelial cells within pustular lesions. The cats were treated with antibiotics and ketoprofen. The cat with progressive dyspnoea was euthanized. The clinical signs in the other cat rapidly and completely resolved following glucocorticoid therapy. It is hypothesized that the reported cases may represent a distinct calicivirus-induced pustular dermatitis following ovariectomy. PMID- 16359308 TI - Generalized calcinosis cutis associated with probable leptospirosis in a dog. AB - A 6.5-year-old male German Shepherd acutely developed renal and hepatic disease. Serology revealed high concentrations of antibodies against Leptospira copenhageni, and a presumptive diagnosis of leptospirosis was made. The dog was successfully treated with antibiotics and supportive care over a 12-day period. Sixty-two days after the initial presentation, alopecia predominantly involving the dorsum and perineal areas developed. The skin lesions expanded over a 20-day period. Histology revealed generalized calcinosis cutis with follicular atrophy. An injection of 0.01 mg kg-1 dexamethasone suppressed serum cortisol concentrations. No treatment was given and lesions resolved over the following 30 days. This is the third case of generalized calcinosis cutis that has developed in an adult dog after severe systemic disease. Both previous cases developed calcinosis cutis in association with blastomycosis. To the authors' knowledge, this is the first report of generalized calcinosis cutis in an adult dog in association with a presumptive bacterial infection. PMID- 16359309 TI - Two cases of FeLV-associated dermatoses. AB - Two cases of feline leukaemia virus (FeLV)-associated dermatosis are described. The first cat was affected by an ulcerative dermatitis identified as a giant-cell dermatosis. The second case was a cutaneous lymphoma. In both cases, FeLV antigens and FeLV genome were demonstrated in the affected skin immunologically and with polymerase chain reaction, respectively. The first case suggests that, like other retroviruses, at least some strains of FeLV can induce syncytium formation. As FeLV antigens and genome were demonstrated in a serologically negative cat, the second case suggests that focal skin FeLV replication may occur. FeLV-associated dermatoses are rare skin conditions that may be under diagnosed. PMID- 16359310 TI - Feline cutaneous and visceral necrotizing panniculitis and steatitis associated with a pancreatic tumour. AB - The association of pancreatic disorders with fat necrosis in domestic animals is rare. This report concerns a case of cutaneous/subcutaneous necrotizing panniculitis and steatitis associated with a pancreatic adenocarcinoma in an 11 year-old male Siamese cat. Clinical investigation revealed variably sized nodules on the trunk, limbs and abdomen. Some of them were ulcerated; others showed a shiny yellow necrotic background featuring irregular sinus tracts. The cat was euthanized at the owner's request before a diagnosis could be made. At necropsy, abundant oily material resembling mustard replaced the subcutaneous tissue and small yellow nodules were disseminated in the omentum, mesentery and serosa of the abdomen. A multilobulated mass arising from the anterior pancreatic head was found along with liver and lymph node metastasis. Histopathology showed wide fistulous tracts draining necrotic fat from the subcutis toward the surface and multifocal areas of necrotic adipocytes replacing the panniculus. Duct-like structures and tubules lined by neoplastic epithelial cells were observed in the primary pancreatic tumour and in the metastatic sites. The aetiology of the fat necrosis was possibly the result of systemic release of lipolytic pancreatic enzymes. PMID- 16359311 TI - Congenital follicular dysplasia in five related coatimundis (Nasua nasua). AB - We report a congenital follicular dysplasia in five coatis from four different litters of the same parents born between 1996 and 2001. These coatis were born apparently alopecic with the entire body covered by very short dark hairs, with secondary lichenification of the skin, crusting and scaling. The main histopathological feature consisted of premature cornification of the cortical cells of the hair shaft. Cells were already fully cornified below the Adamson's fringe, leading to a disorganized, fragmented and constricted hair shaft. Based on the history of the animals and the nature of the lesions, a genetic defect in hair shaft keratinization was suspected. PMID- 16359312 TI - Equine cutaneous non-neoplastic nodular and proliferative lesions in the Pacific Northwest. AB - A retrospective study examined data on cutaneous nodular and proliferative lesions in horses, donkeys and mules submitted to the Veterinary Diagnostic Laboratory at Oregon State University in a 3.5-year period. One hundred and sixteen non-neoplastic nodular and proliferative lesions were identified, comprising 18% of all lesions studied and 6.4% of total equine pathology accessions. Exuberant granulation tissue, eosinophilic granuloma, fungal granuloma, cysts and habronaemiasis were most common, and constituted 91% of non neoplastic lesions, 16% of all lesions studied, and 5.85% of total equine pathology accessions. Calcinosis circumscripta and cutaneous granulomas, including eosinophilic granuloma, fungal granuloma, and other granulomas, were most common in horses up to 10 years of age. Eosinophilic granuloma was more common in males than in females, and was diagnosed most often in autumn and winter. Fungal granuloma was common in this population and was most often diagnosed in the spring. Habronaemiasis was diagnosed in late summer and early autumn, and was most common in males. Results of this study indicate that equine non-neoplastic nodular and proliferative skin lesions are commonly submitted for histopathologic examination. Results also suggest that regional differences exist regarding incidence of cutaneous fungal granuloma and season of occurrence of eosinophilic granuloma in horses. PMID- 16359313 TI - Domains of group A streptococcal M protein that confer resistance to phagocytosis, opsonization and protection: implications for vaccine development. AB - Streptococcus pyogenes (group A streptococcus) colonizes skin and throat tissues resulting in a range of benign and serious human diseases. Opsonization and phagocytosis are important defence mechanisms employed by the host to destroy group A streptococci. Antisera against the cell-surface M protein, of which over 150 different types have been identified, are opsonic and contribute to disease protection. In this issue of Molecular Microbiology, Sandin and colleagues have comprehensively analysed the regions of M5 protein that contribute to phagocytosis resistance and opsonization. Human plasma proteins bound to M5 protein B- and C-repeats were shown to block opsonization, an observation that needs to be carefully considered for the development of M protein-derived vaccines. While safe and efficacious human group A streptococcal vaccines are not commercially available, candidate M protein-derived vaccines have shown promise in murine vaccine models and a recent phase 1 human clinical trial. PMID- 16359314 TI - House cleaning, a part of good housekeeping. AB - Cellular metabolism constantly generates by-products that are wasteful or even harmful. Such compounds are excreted from the cell or are removed through hydrolysis to normal cellular metabolites by various 'house-cleaning' enzymes. Some of the most important contaminants are non-canonical nucleoside triphosphates (NTPs) whose incorporation into the nascent DNA leads to increased mutagenesis and DNA damage. Enzymes intercepting abnormal NTPs from incorporation by DNA polymerases work in parallel with DNA repair enzymes that remove lesions produced by modified nucleotides. House-cleaning NTP pyrophosphatases targeting non-canonical NTPs belong to at least four structural superfamilies: MutT-related (Nudix) hydrolases, dUTPase, ITPase (Maf/HAM1) and all-alpha NTP pyrophosphatases (MazG). These enzymes have high affinity (Km's in the micromolar range) for their natural substrates (8-oxo-dGTP, dUTP, dITP, 2-oxo-dATP), which allows them to select these substrates from a mixture containing a approximately 1000-fold excess of canonical NTPs. To date, many house-cleaning NTPases have been identified only on the basis of their side activity towards canonical NTPs and NDP derivatives. Integration of growing structural and biochemical data on these superfamilies suggests that their new family members cleanse the nucleotide pool of the products of oxidative damage and inappropriate methylation. House-cleaning enzymes, such as 6-phosphogluconolactonase, are also part of normal intermediary metabolism. Genomic data suggest that house-cleaning systems are more abundant than previously thought and include numerous analogous enzymes with overlapping functions. We discuss the structural diversity of these enzymes, their phylogenetic distribution, substrate specificity and the problem of identifying their true substrates. PMID- 16359315 TI - Binding of human plasma proteins to Streptococcus pyogenes M protein determines the location of opsonic and non-opsonic epitopes. AB - Antibodies directed against a pathogenic microorganism may recognize either protective or non-protective epitopes. Because antibodies elicited by a vaccine must be directed against protective epitopes, it is essential to understand the molecular properties that distinguish the two types of epitope. Here we analyse this problem for the antiphagocytic M protein of Streptococcus pyogenes, using the opsonizing capacity of antibodies to estimate their ability to confer protection in vivo. Our studies were focused on the M5 protein, which has three surface-exposed regions: the amino-terminal hypervariable region (HVR) and the B- and C-repeat regions. We first analysed the role of different M5 regions in phagocytosis resistance under non-immune conditions, employing chromosomal mutants expressing M5 proteins with internal deletions, and demonstrate that only the B-repeat region is essential for phagocytosis resistance. However, only antibodies to the HVR were opsonic. This apparent paradox could be explained by the ability of fibrinogen and albumin to specifically bind to the B- and C repeats, respectively, causing inhibition of antibody binding under physiological conditions, while antibodies to the HVR could bind and promote deposition of complement. These data indicate that binding of human plasma proteins plays an important role in determining the location of opsonic and non-opsonic epitopes in streptococcal M protein. PMID- 16359316 TI - Tetratricopeptide repeats in the type III secretion chaperone, LcrH: their role in substrate binding and secretion. AB - Non-flagellar type III secretion systems (T3SSs) transport proteins across the bacterial cell and into eukaryotic cells. Targeting of proteins into host cells requires a dedicated translocation apparatus. Efficient secretion of the translocator proteins that make up this apparatus depends on molecular chaperones. Chaperones of the translocators (also called class-II chaperones) are characterized by the possession of three tandem tetratricopeptide repeats (TPRs). We wished to dissect the relations between chaperone structure and function and to validate a structural model using site-directed mutagenesis. Drawing on a number of experimental approaches and focusing on LcrH, a class-II chaperone from the Yersinia Ysc-Yop T3SS, we examined the contributions of different residues, residue classes and regions of the protein to chaperone stability, chaperone substrate binding, substrate stability and secretion and regulation of Yop protein synthesis. We confirmed the expected role of the conserved canonical residues from the TPRs to chaperone stability and function. Eleven mutations specifically abrogated YopB binding or secretion while three mutations led to a specific loss of YopD secretion. These are the first mutations described for any class-II chaperone that allow interactions with one translocator to be dissociated from interactions with the other. Strikingly, all mutations affecting the interaction with YopB mapped to residues with side chains projecting from the inner, concave surface of the modelled TPR structure, defining a YopB interaction site. Conversely, all mutations preventing YopD secretion affect residues that lie on the outer, convex surface of the triple-TPR cluster in our model, suggesting that this region of the molecule represents a distinct interaction site for YopD. Intriguingly, one of the LcrH double mutants, Y40A/F44A, was able to maintain stable substrates inside bacteria, but unable to secrete them, suggesting that these two residues might influence delivery of substrates to the secretion apparatus. PMID- 16359317 TI - Differential effects of chemoreceptor methylation-domain mutations on swarming and development in the social bacterium Myxococcus xanthus. AB - The soil bacterium Myxococcus xanthus is a model organism for the study of multicellular behaviour and development in bacteria. M. xanthus cells move on solid surfaces by gliding motility, periodically reversing their direction of movement. Motility is co-ordinated to allow cells to effectively feed on macromolecules or prey bacteria when nutrients are plentiful and to form developmental fruiting bodies when nutrients are limiting. The Frz signal transduction pathway regulates cellular movements by modulating cell reversal frequency. Input to the Frz pathway is controlled by the cytoplasmic receptor, FrzCD, a methyl-accepting chemotaxis protein (MCP). FrzCD lacks the transmembrane and periplasmic domains common to MCPs but contains a unique N-terminal domain, the predicted ligand-binding domain. As deletion of the N-terminal domain of FrzCD only results in minor defects in motility, we investigated the possibility that the methylation of the conserved C-terminal domain of FrzCD plays a central role in regulating the pathway. For this study, each of the potential methylation sites of FrzCD were systematically modified by site-directed mutagenesis, substituting glutamine/glutamate pairs for alanines. Four of the seven mutations produced dramatic phenotypes; two of the mutations had a stimulatory effect on the pathway, as evidenced by cells hyper-reversing, whereas another two had an inhibitory effect, causing these cells to rarely reverse. These four mutants displayed defects in vegetative swarming and developmental aggregation. These results suggests a model in which the methylation domain can both activate and inhibit the Frz pathway depending on which residues are methylated. The diversity of phenotypes suggests that specific modifications of FrzCD act to differentially regulate motility and developmental aggregation in M. xanthus. PMID- 16359318 TI - Characterization of a Methanosarcina acetivorans mutant unable to translate UAG as pyrrolysine. AB - The methyltransferases initiating methanogenesis from trimethylamine, dimethylamine and monomethylamine possess a novel residue, pyrrolysine. Pyrrolysine is the 22nd amino acid, because it is encoded by a single amber (UAG) codon in methylamine methyltransferase transcripts. A dedicated tRNA(CUA) for pyrrolysine, tRNA(Pyl), is charged by a pyrrolysyl-tRNA synthetase with pyrrolysine. As the first step towards the genetic analysis of UAG translation as pyrrolysine, a 761 base-pair genomic segment in Methanosarcina acetivorans containing the pylT gene (encoding tRNA(Pyl)) was deleted and replaced by a puromycin resistance cassette. The DeltappylT mutant lacks detectable tRNA(Pyl), but grows as wild-type on methanol or acetate. Unlike wild-type, the DeltappylT strain cannot grow on any methylamine, nor use monomethylamine as sole nitrogen source. Wild-type cells, but not DeltappylT, have monomethylamine methyltransferase activity during growth on methanol. Immunoblot analysis indicated monomethylamine methyltransferase was absent in DeltappylT. The phenotype of DeltappylT reveals the deficiency in methylamine metabolism expected of a Methanosarcina species unable to decode UAG codons as pyrrolysine, but also that loss of pylT does not compromise growth on other substrates. These results indicate that in-depth genetic analysis of UAG translation as pyrrolysine is feasible, as deletion of pylT is conditionally lethal depending on growth substrate. PMID- 16359319 TI - The polysaccharide capsule of the pathogenic fungus Cryptococcus neoformans enlarges by distal growth and is rearranged during budding. AB - The capsule of Cryptococcus neoformans can undergo dramatic enlargement, a phenomenon associated with virulence. A prior study that used Ab to the capsule as a marker for older capsular material concluded that capsule growth involved the intermixing of new and old capsular material with displacement of older capsular polysaccharide towards the surface. Here we have revisited that question using complement (C), which binds to capsular polysaccharide covalently, and cannot redistribute by dissociation and binding at different sites. The experimental approach involved binding of C to cells with small capsules, inducing capsule growth, and following the location of C relative to the cell wall as the capsule enlarged. C remained close to the cell wall during capsule growth, indicating that capsule enlargement occurred by addition of new polysaccharide near the capsule edge. This conclusion was confirmed by an independent method that employed radioactive metabolic labelling of newly synthesized capsule with 3H-mannose followed by gradual capsular stripping with gamma-radiation. Capsule growth proceeded to a certain size, which was a function of cell size, and was not degraded when the cells were transferred to a non inducing medium. During budding, an opening appeared in the capsule of the mother cell that permitted the nascent bud to separate. Scanning EM suggested that a physical separation formed between the capsules of the mother and daughter cells during budding, which may avoid mixture between both capsules. Our results indicate that C. neoformans capsular enlargement also occurs by apical growth and that budding results in capsular rearrangements. PMID- 16359320 TI - Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation. AB - The culturability of several actinobacteria is controlled by resuscitation promoting factors (Rpfs). These are proteins containing a c. 70-residue domain that adopts a lysozyme-like fold. The invariant catalytic glutamate residue found in lysozyme and various bacterial lytic transglycosylases is also conserved in the Rpf proteins. Rpf from Micrococcus luteus, the founder member of this protein family, is indeed a muralytic enzyme, as revealed by its activity in zymograms containing M. luteus cell walls and its ability to (i) cause lysis of Escherichia coli when expressed and secreted into the periplasm; (ii) release fluorescent material from fluorescamine-labelled cell walls of M. luteus; and (iii) hydrolyse the artificial lysozyme substrate, 4-methylumbelliferyl-beta-D-N,N',N'' triacetylchitotrioside. Rpf activity was reduced but not completely abolished when the invariant glutamate residue was altered. Moreover, none of the other acidic residues in the Rpf domain was absolutely required for muralytic activity. Replacement of one or both of the cysteine residues that probably form a disulphide bridge within Rpf impaired but did not completely abolish muralytic activity. The muralytic activities of the Rpf mutants were correlated with their abilities to stimulate bacterial culturability and resuscitation, consistent with the view that the biological activity of Rpf results directly or indirectly from its ability to cleave bonds in bacterial peptidoglycan. PMID- 16359321 TI - Release of the type I secreted alpha-haemolysin via outer membrane vesicles from Escherichia coli. AB - The alpha-haemolysin is an important virulence factor commonly expressed by extraintestinal pathogenic Escherichia coli. The secretion of the alpha haemolysin is mediated by the type I secretion system and the toxin reaches the extracellular space without the formation of periplasmic intermediates presumably in a soluble form. Surprisingly, we found that a fraction of this type I secreted protein is located within outer membrane vesicles (OMVs) that are released by the bacteria. The alpha-haemolysin appeared very tightly associated with the OMVs as judged by dissociation assays and proteinase susceptibility tests. The alpha haemolysin in OMVs was cytotoxically active and caused lysis of red blood cells. The OMVs containing the alpha-haemolysin were distinct from the OMVs not containing alpha-haemolysin, showing a lower density. Furthermore, they differed in protein composition and one component of the type I secretion system, the TolC protein, was found in the lower density vesicles. Studies of natural isolates of E. coli demonstrated that the localization of alpha-haemolysin in OMVs is a common feature among haemolytic strains. We propose an alternative pathway for the transport of the type I secreted alpha-haemolysin from the bacteria to the host cells during bacterial infections. PMID- 16359322 TI - Identification of the central quorum sensing regulator of virulence in the enteric phytopathogen, Erwinia carotovora: the VirR repressor. AB - In the Gram-negative phytopathogen, Erwinia carotovora ssp. atroseptica (Eca) virulence depends on the production of a N-(3-oxohexanoyl)-L-homoserine lactone (OHHL) quorum sensing (QS) signal. This work identifies the elusive 'missing link' between QS and virulence in Erwinia. We have identified and characterized a novel regulator of virulence, VirR, in Eca and show that a virR mutation completely restores virulence factor production to an Eca mutant unable to synthesize OHHL. This effect of the virR mutation translates to a restoration of virulence to wild-type levels and thus provides evidence that VirR acts to prevent the production of virulence factors at low cell density. We also show that, in Eca, transcription of virulence genes is controlled by OHHL and that this control is effected through the action of VirR. We also demonstrate that the VirR regulatory pathway is present and functional in both blackleg and soft rotting species of Erwinia. PMID- 16359323 TI - Virulence and drug resistance roles of multidrug efflux systems of Salmonella enterica serovar Typhimurium. AB - Drug efflux systems play a major role in resistance to a wide range of noxious compounds in several Gram negative species. Here, we report the drug resistance and virulence phenotypes of Salmonella mutants defective in either resistance nodulation-division (RND)-type systems and/or in drug efflux systems belonging to the major facilitator (MFS), multidrug and toxic compound extrusion (MATE), and ATP-binding cassette (ABC) superfamilies. We determined that nine potential drug transporters contribute to drug resistance of Salmonella and found that the Salmonella-specific MdsABC system conferred resistance to a variety of toxic compounds. The RND-type MdsAB system could function with either MdsC, which is encoded in the same operon, or TolC as the outer membrane component. Although the Salmonella EmrAB, MdfA and MdtK are 90% identical in their amino acid sequences to their Escherichia coli homologues, the drug specificity of Salmonella transporters was different from that reported for equivalent E. coli transporters. Deletion of the macAB genes attenuated Salmonella virulence and a strain lacking all drug efflux systems was avirulent when mice were inoculated by the oral route. The promoter region of the macAB drug efflux system genes harbours a binding site for the response regulator PhoP, which functions to repress macAB transcription. The PhoP/PhoQ two-component system is a major regulator of Salmonella virulence, which underscores the connection between drug efflux systems and virulence. PMID- 16359324 TI - Mucin-Pseudomonas aeruginosa interactions promote biofilm formation and antibiotic resistance. AB - Pseudomonas aeruginosa is an opportunistic pathogen that causes chronic lung infections in people suffering from cystic fibrosis (CF). In CF airways, P. aeruginosa forms surface-associated communities called biofilms. Compared with free-swimming cultures, biofilms resist clearance by the host immune system and display increased resistance to antimicrobial agents. In this study we developed a technique to coat surfaces with molecules that are abundant in CF airways in order to investigate their impact on P. aeruginosa biofilm development. We found that P. aeruginosa biofilm development proceeds differently on surfaces coated with the glycoprotein mucin compared with biofilm development on glass and surfaces coated with actin or DNA. Biofilms formed on mucin-coated surfaces developed large cellular aggregates and had increased tolerance to the antibiotic tobramycin compared with biofilms grown on glass. Analysis of selected mutant backgrounds in conjunction with time-lapse microscopy revealed that surface associated motility was blocked on the mucin surface. Furthermore, our data suggest that a specific adhesin-mucin interaction immobilizes the bacterium on the surface. Together, these experiments suggest that mucin, which may serve as an attachment surface in CF airways, impacts P. aeruginosa biofilm development and function. PMID- 16359325 TI - Porphyromonas gingivalis-induced platelet aggregation in plasma depends on Hgp44 adhesin but not Rgp proteinase. AB - Evidence from recent epidemiological studies suggests a link between periodontal infections and increased risk of atherosclerosis and related cardiovascular and cerebrovascular events in human subjects. One of the major pathogens of periodontitis, Porphyromonas gingivalis, has the ability to aggregate human platelets in platelet-rich plasma (PRP). Mechanism of P. gingivalis-induced platelet aggregation in PRP was investigated. Proteinase inhibitors toward Arg gingipain (Rgp) and Lys-gingipain (Kgp) did not suppress P. gingivalis-induced platelet aggregation in PRP, whereas the Rgp inhibitor markedly inhibited P. gingivalis-induced platelet aggregation using washed platelets. Mutant analysis revealed that P. gingivalis-induced platelet aggregation in PRP depended on Rgp-, Kgp- and haemagglutinin A (HagA)-encoding genes that intragenically coded for adhesins such as Hgp44. Hgp44 adhesin on the bacterial cell surface, which was processed by Rgp and Kgp proteinases, was essential for P. gingivalis-induced platelet aggregation in PRP. P. gingivalis cell-reactive IgG in plasma, and FcgammaRIIa receptor and to a lesser extent GPIbalpha receptor on platelets were found to be a prerequisite for P. gingivalis-induced platelet aggregation in PRP. These results reveal a novel mechanism of platelet aggregation by P. gingivalis. PMID- 16359326 TI - Protein-induced DNA bending clarifies the architectural organization of the sigma54-dependent glnAp2 promoter. AB - Sigma54-RNA polymerase (Esigma54) predominantly contacts one face of the DNA helix in the closed promoter complex, and interacts with the upstream enhancer bound activator via DNA looping. Up to date, the precise face of Esigma54 that contacts the activator to convert the closed complex to an open one remains unclear. By introducing protein-induced DNA bends at precise locations between upstream enhancer sequences and the core promoter of the sigma54-dependent glnAp2 promoter without changing the distance in-between, we observed a strong enhanced or decreased promoter activity, especially on linear DNA templates in vitro. The relative positioning and orientations of Esigma54, DNA bending protein and enhancer-bound activator on linear DNA were determined by in vitro footprinting analysis. Intriguingly, the locations from which the DNA bending protein exerted its optimal stimulatory effects were all found on the opposite face of the DNA helix compared with the DNA bound Esigma54 in the closed complex. Therefore, these results provide evidence that the activator must approach the Esigma54 closed complexes from the unbound face of the promoter DNA helix to catalyse open complex formation. This proposal is further supported by the modelling of activator-promoter DNA-Esigma54 complex. PMID- 16359327 TI - Characterization of the putative operon containing arylamine N-acetyltransferase (nat) in Mycobacterium bovis BCG. AB - Mycobacterium bovis BCG and Mycobacterium tuberculosis possess a single arylamine N-acetyltransferase whose gene is predicted to occur within a six-gene operon. Deletion of the nat gene caused an extended lag phase in M. bovis BCG and a cell morphology associated with an altered pattern of cell wall mycolates. Analysis of cDNA from M. bovis BCG shows that during in vitro growth all the genes in the putative nat operon are expressed and the open reading frames are contiguous, supporting the existence of an operon. Two genes in the operon, Mb3599c and Mb3600c, are predicted to encode homologues of enzymes annotated as a 2,3 dihydroxybiphenyl 1,2-dioxygenase (bphC5) and a 2-hydroxy-6-oxo-6-phenylhexa-2,4 dienoate hydrolase (bphD2), respectively, in Rhodococcus RHA1. As predicted, M. bovis BCG cell lysates metabolized the BphC substrate 2,3-dihydroxybiphenyl (2,3 DHB) to 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA), a BphD substrate, which was subsequently hydrolysed. Immunoprecipitation of the BphD homologue from these lysates led to an accumulation of HOPDA. M. bovis BCG growth on both solid and liquid media was inhibited with either 2,3-DHB or an inhibitor of BphC, 3 chlorocatechol (3-CC). In addition, incubation with 2,3-DHB affects the lipid composition of the cell wall resulting in a diminished level of mycolates and an altered cell morphology similar to the Deltanat strain. We propose the enzymes encoded by the putative operon have a similar endogenous role to that of the NAT enzyme and are part of a pathway important for cell wall synthesis. PMID- 16359328 TI - Bile acids stimulate biofilm formation in Vibrio cholerae. AB - Vibrio cholerae is a Gram-negative bacterium that causes the acute diarrhoeal disease cholera. After the bacterium is ingested, it passes through the digestive tract, encountering various environmental stresses including the acidic milieu of the stomach and the toxic effects of bile in the duodenum. While these stresses serve as part of a host defence system, V. cholerae has evolved resistance mechanisms that allow it to evade these defences and establish infection. We examined the expression profiles of V. cholerae in response to bile or bile acids and found an induction of biofilm genes. We found that V. cholerae shows significantly enhanced biofilm formation in response to bile acids, and that bacteria within the biofilm are more resistant to the toxicity of bile acids compared with planktonic cells. Bile acid induction of biofilms was found to be dependent on the vps genes (Vibrio polysaccharide synthesis) and their transcriptional activator VpsR, but VpsT is not required. These results contribute to the developing picture of a complex relationship between V. cholerae and its environment within the host during infection. PMID- 16359329 TI - A likely molecular basis of the susceptibility of Giardia lamblia towards oxygen. AB - Giardia lamblia is an amitochondrial protozoan susceptible to oxygen, but the molecular basis for it remains unclear. A Giardia NAD(P)H:menadione oxidoreductase (DT-diaphorase) is known to catalyse a single electron transfer reaction with quinones as the likely two-electron acceptor when oxygen is absent. Here we overexpressed this enzyme in Giardia trophozoites and observed a significantly enhanced susceptibility of the cells towards oxygen. A knock-down of this enzyme resulted, however, in more oxygen-tolerant Giardia cells growing equally well under anaerobic and aerobic conditions. The function of DT diaphorase could be thus a major, if not the only, cause for the oxygen susceptibility of Giardia. Overexpressed DT-diaphorase is accompanied by increased intracellular hydrogen peroxide. An overexpression of Fe-superoxide dismutase in Giardia led also to a similarly heightened sensitivity to oxygen. Thus, generation of H2O2 from superoxide anion likely produced from DT-diaphorase catalysed reaction using oxygen as electron acceptor may constitute the molecular basis for Giardia susceptibility to oxygen. A functional homologue of DT diaphorase in Giardia, NADH oxidase, uses oxygen as the preferred electron acceptor and reduces it to water. Overexpression of this enzyme in Giardia resulted in significantly enhanced growth under aerobic conditions. Giardia NADH oxidase could be thus an instrumental enzyme for the organism to adapt to and to tolerate an aerobic living environment. PMID- 16359330 TI - Fibronectin-binding proteins of Staphylococcus aureus mediate activation of human platelets via fibrinogen and fibronectin bridges to integrin GPIIb/IIIa and IgG binding to the FcgammaRIIa receptor. AB - Staphylococcus aureus is a leading cause of infective endocarditis (IE). Platelet activation promoted by S. aureus resulting in aggregation and thrombus formation is an important step in the pathogenesis of IE. Here, we report that the fibrinogen/fibronectin-binding proteins FnBPA and FnBPB are major platelet activating factors on the surface of S. aureus from the exponential phase of growth. Truncated derivatives of FnBPA, presenting either the fibrinogen-binding A domain or the fibronectin-binding BCD region, each promoted platelet activation when expressed on the surface of S. aureus or Lactococcus lactis, indicating two distinct mechanisms of activation. FnBPA-promoted platelet activation is mediated by fibrinogen and fibronectin bridges between the A domain and the BCD domains, respectively, to the low affinity form of the integrin GPIIb/IIIa on resting platelets. Antibodies recognizing the FnBPA A domain or the complex between the FnBPA BCD domains and fibronectin were essential for activation promoted by bacteria expressing the A domain or the BCD domain respectively. Activation was inhibited by a monoclonal antibody (IV-3) specific for the FcgammaRIIa IgG receptor on platelets. We propose that the activation of quiescent platelets by bacteria expressing FnBPs involves the formation of a bridge between the bacterial cell and the platelet surface by (i) fibronectin and fibrinogen interacting with the low affinity form of GPIIb/IIIa and (ii) by antibodies specific to FnBPs that engage the platelet Fc receptor FcgammaRIIa. Platelet activation by S. aureus clinical IE isolates from both the exponential and stationary phases of growth was completely inhibited by monoclonal antibody IV-3 suggesting that the IgG-FcgammaRIIa interaction is of fundamental importance for platelet activation mediated by this organism. This suggests new avenues for development of therapeutics against vascular infections. PMID- 16359331 TI - Remodelling of the Escherichia coli outer membrane by two small regulatory RNAs. AB - Small non-coding RNAs that play important regulatory roles exist in numerous organisms. In Escherichia coli, about 60 small RNAs have been found and those that have been studied are involved in the response and adaptation to different stresses. RygA and RygB, two of these small RNAs, were identified on the basis of their conservation between different species and their ability to bind Hfq. They are adjacent on the chromosome and have sequence similarity at their 5' and 3' ends but distinct central regions, suggesting that they could regulate the expression of both common and distinct genes. A screen using a multicopy E. coli library led to identification of the response regulator OmpR and its associated sensor kinase EnvZ as positive regulators of rygA and rygB transcription. Therefore, RygA and RygB were renamed OmrA and OmrB respectively (for OmpR regulated sRNAs A and B). When expressed at high levels, OmrA and OmrB RNAs negatively regulate the expression of several genes encoding multiple outer membrane proteins, including cirA, fecA, fepA and ompT. Taken together, these data suggest that OmrA and OmrB participate in the regulation of outer membrane composition in response to environmental conditions. PMID- 16359332 TI - The chaperone binding domain of SopE inhibits transport via flagellar and SPI-1 TTSS in the absence of InvB. AB - Type III secretion systems (TTSS) are used by many Gram-negative pathogens for transporting effector proteins into eukaryotic host cells. Two modes of type III effector protein transport can be distinguished: transport into the surrounding medium (secretion) and cell-contact induced injection of effector proteins directly into the host cell cytosol (translocation). Two domains within the N terminal regions of effector proteins determine the mode of transport. The amino terminal approximately 20 amino acids (N-terminal secretion signal, NSS) mediate secretion. In contrast, translocation generally requires the NSS, the adjacent approximately 100 amino acids (chaperone binding domain, CBD) and binding of the cognate chaperone to this CBD. TTSS are phylogenetically related to flagellar systems. Because both systems are expressed in Salmonella Typhimurium, correct effector protein transport involves at least two decisions: transport via the Salmonella pathogenicity island 1 (SPI-1) but not the flagellar TTSS (= specificity) and translocation into the host cell instead of secretion into the surrounding media (= transport mode). The mechanisms guiding these decisions are poorly understood. We have studied the S. Typhimurium effector protein SopE, which is specifically transported via the SPI-1 TTSS. Secretion and translocation strictly require the cognate chaperone InvB. Alanine replacement of amino acids 30-42 (and to some extent 44-54) abolished tight InvB binding, abolished translocation into the host cell and led to secretion of SopE via both, the flagellar and the SPI-1 TTSS. In clear contrast to wild-type SopE, secretion of SopE(Ala30-42) and SopE(Ala44-54) via the SPI-1 and the flagellar export system did not require InvB. These data reveal a novel function of the CBD: the CBD inhibits secretion of wild-type SopE via the flagellar and the SPI-1 TTSS in the absence of the chaperone InvB. Our data provide new insights into mechanisms ensuring specific effector protein transport by TTSS. PMID- 16359333 TI - Involvement of the Escherichia coli folate-binding protein YgfZ in RNA modification and regulation of chromosomal replication initiation. AB - The Escherichia coli hda gene codes for a DnaA-related protein that is essential for the regulatory inactivation of DnaA (RIDA), a system that controls the initiation of chromosomal replication. We have identified the ygfZ gene, which encodes a folate-binding protein, as a suppressor of hda mutations. The ygfZ null mutation suppresses an hda null mutation. The over-initiation and abortive elongation phenotypes conferred by the hda mutations are partially suppressed in an hda ygfZ background. The accumulation of the active form of DnaA, ATP-DnaA, in the hda mutant is suppressed by the disruption of the ygfZ gene, indicating that YgfZ is involved in regulating the level of ATP-DnaA. Although ygfZ is not an essential gene, the ygfZ disruptant grows slowly, especially at low temperature, demonstrating that this gene is important for cellular proliferation. We have identified mnmE (trmE) as a suppressor of the ygfZ disruption. This gene encodes a GTPase involved in tRNA modification. Examination of RNA modification in the ygfZ mutant reveals reduced levels of 2-methylthio N(6)-isopentenyladenosine [corrected] indicating that YgfZ participates in the methylthio-group formation of this modified nucleoside in some tRNAs. These results suggest that YgfZ is a key factor in regulatory networks that act via tRNA modification. PMID- 16359334 TI - P1 plasmid partition: in vivo evidence for the ParA- and ParB-mediated formation of an anchored parS complex in the absence of a partner parS. AB - ParA and ParB proteins and cis-acting site, parS, are required to partition plasmid P1 faithfully to daughter cells. The process may initiate from plasmids paired by ParB at which recruited ParA then acts to effect the separation. We previously reported evidence for ParB-mediated pairing of parS sites on plasmids in the absence of ParA. In DNA gyrase-inhibited cells, the pairing prevented diffusion of transcription-generated positive supercoils. This supercoil trapping was almost entirely in plasmid dimers, where the location of the parS sites in cis facilitated their pairing. Here we show that the addition of ParA blocked supercoil diffusion also in plasmid monomers. The possibility that this result is attributed to an enhancement by ParA of ParB-mediated pairing in trans is consistent with our finding that ParA appeared to partially suppress the pairing defect of two mutant ParB proteins. However, enhanced pairing alone could not account for the diffusion barrier in plasmid monomers; it was manifest in monomers even when they were largely devoid of partners in the same cell. Apparently, ParA altered the ParB-parS complex such that it could no longer swivel, most likely by anchoring it, a reaction of probable relevance to partition. PMID- 16359335 TI - Bacteroides fragilis mobilizable transposon Tn5520 requires a 71 base pair origin of transfer sequence and a single mobilization protein for relaxosome formation during conjugation. AB - Tn5520 is the smallest known bacterial mobilizable transposon and was isolated from an antibiotic resistant Bacteroides fragilis clinical isolate. When a conjugation apparatus is provided in trans, Tn5520 is mobilized (transferred) efficiently within, and from, both Bacteroides spp. and Escherichia coli. Only two genes are present on Tn5520; one encodes an integrase, and the other a multifunctional mobilization (Mob) protein BmpH. BmpH is essential for Tn5520 mobility. The focus of this study was to identify the Tn5520 origin of conjugative transfer (oriT) and to study BmpH-oriT binding. We delimited the functional Tn5520 oriT to a 71 bp sequence upstream of the bmpH gene. A plasmid vector harbouring this minimal 71 bp oriT was mobilized at the same frequency as that of intact Tn5520. The minimal oriT contains one 17 bp inverted repeat (IR) sequence. We constructed and tested multiple IR mutants and showed that the IR was essential in its entirety for mobilization. A nick site sequence (5'-GCTAC 3') was also identified within the minimal oriT; this sequence resembled nick sites found in plasmids of Gram positive origin. We further showed that mutation of a highly conserved GC dinucleotide in the nick site sequence completely abolished mobilization. We also purified BmpH and showed that it specifically bound a Tn5520 oriT fragment in electrophoretic mobility shift assays. We also identified non-nick site sequences within the minimal oriT that were essential for mobilization. We hypothesize that transposon-based single Mob protein systems may contribute to efficient gene dissemination from Bacteroides spp., because fewer DNA processing proteins are required for relaxosome formation. PMID- 16359337 TI - Mutagenesis via IS transposition in Deinococcus radiodurans. AB - Analysis of the complete genome indicates that insertion sequences (ISs) are abundant in the radio-resistant bacterium Deinococcus radiodurans. By developing a forward mutagenesis assay to detect any inactivation events in D. radiodurans, we found that in the presence of an active mismatch repair system 75% of the mutations to trimethoprim-resistance (Tmp(R)) resulted from an IS insertion into the thyA coding region. Analysis of their distribution among the spontaneous Tmp(R) mutants indicated that five different ISs were transpositionally active. A type II Miniature Inverted-repeat Transposable Element (MITE), related to one of the deinococcal ISs, was also discovered as an insertion into thyA. Seven additional genomic copies of this MITE element were identified by BLASTN. Gamma ray irradiation of D. radiodurans led to an increase of up to 10-fold in the frequency of Tmp(R) mutants. Analysis of the induced mutations in cells exposed to 10 kGy indicated that gamma-irradiation induced transposition of ISDra2 approximately 100-fold. A 50-fold induction of ISDra2 transposition was also observed in cells exposed to 600 J m(-2) UV-irradiation. Point mutations to rifampicin resistance (Rif(R)) were also induced by gamma-irradiation to reach a plateau at 2 kGy. The plateau value represented a 16-fold increase in the mutant frequency over the background. Although error-free repair strategies predominate in D. radiodurans, an upregulation of transposition, as well as induction of point mutations in cells recovering from DNA damage, provide a genetic variability that may have long-term evolutionary consequences on the fitness of this organism in its habitat. PMID- 16359336 TI - Dissection of functional domains of the polar localization factor PodJ in Caulobacter crescentus. AB - The polar organelle development protein, PodJ, is important for proper establishment of polarity in Caulobacter crescentus. podJ null mutants are unable to form holdfast or pili, have reduced swarming motility, and have difficulty ejecting the flagellum during the swarmer to stalked cell transition. In this study, we create a series of truncation mutants to investigate functional domains of PodJ. We show that PodJ has a transmembrane domain between amino acids 600 and 670. We identify a periplasmic region important for pili production and a cytoplasmic region required for holdfast formation and swarming motility, and establish that PleC localization is not required for holdfast formation and motility in soft agar. Analysis of the mutants reveals that the last 54 amino acids of the protein negatively regulate processing of the full-length form of the protein, PodJ(L), to a shorter form, PodJ(S). Finally, we identify a cytoplasmic region of PodJ involved in targeting it to the flagellar pole, and a periplasmic region required for localization of PleC. PMID- 16359338 TI - ADP-ribosylation of dinitrogenase reductase in Azospirillum brasilense is regulated by AmtB-dependent membrane sequestration of DraG. AB - Nitrogen fixation in some diazotrophic bacteria is regulated by mono-ADP ribosylation of dinitrogenase reductase (NifH) that occurs in response to addition of ammonium to the extracellular medium. This process is mediated by dinitrogenase reductase ADP-ribosyltransferase (DraT) and reversed by dinitrogenase reductase glycohydrolase (DraG), but the means by which the activities of these enzymes are regulated are unknown. We have investigated the role of the P(II) proteins (GlnB and GlnZ), the ammonia channel protein AmtB and the cellular localization of DraG in the regulation of the NifH-modification process in Azospirillum brasilense. GlnB, GlnZ and DraG were all membrane associated after an ammonium shock, and both this membrane sequestration and ADP ribosylation of NifH were defective in an amtB mutant. We now propose a model in which membrane association of DraG after an ammonium shock creates a physical separation from its cytoplasmic substrate NifH thereby inhibiting ADP-ribosyl removal. Our observations identify a novel role for an ammonia channel (Amt) protein in the regulation of bacterial nitrogen metabolism by mediating membrane sequestration of a protein other than a P(II) family member. They also suggest a model for control of ADP-ribosylation that is likely to be applicable to all diazotrophs that exhibit such post-translational regulation of nitrogenase. PMID- 16359339 TI - Limited concentration of RecA delays DNA double-strand break repair in Deinococcus radiodurans R1. AB - To evaluate the importance of RecA in DNA double-strand break (DSB) repair, we examined the effect of low and high RecA concentrations such as 2500 and 100 000 molecules per cell expressed from the inducible Pspac promoter in Deinococcus radiodurans in absence or in presence of IPTG respectively. We showed that at low concentration, RecA has a negligible effect on cell survival after gamma irradiation when bacteria were immediately plated on TGY agar whereas it significantly decreased the survival to gamma-irradiation of DeltaddrA cells while overexpression of RecA can partially compensate the loss of DdrA protein. In contrast, when cells expressing limited concentration of RecA were allowed to recover in TGY2X liquid medium, they showed a delay in mending DSB, failed to reinitiate DNA replication and were committed to die during incubation. A deletion of irrE resulted in sensitivity to gamma-irradiation and mitomycin C treatment. Interestingly, constitutive high expression of RecA compensates partially the DeltairrE sensitization to mitomycin C. The cells with low RecA content also failed to cleave LexA after DNA damage. However, neither a deletion of the lexA gene nor the expression of a non-cleavable LexA(Ind-) mutant protein had an effect on survival or kinetics of DNA DSB repair compared with their lexA+ counterparts in recA+ as well as in bacteria expressing limiting concentration of RecA, suggesting an absence of relationship between the absence of LexA cleavage and the loss of viability or the delay in the kinetics of DSB repair. Thus, LexA protein seems to play no major role in the recovery processes after gamma irradiation in D. radiodurans. PMID- 16359340 TI - Induction of the heat shock regulon in response to increased mistranslation requires oxidative modification of the malformed proteins. AB - The Escherichia coli rpsD12 allele, which reduces translational fidelity and elevates expression of heat shock protein (Hsp) genes, only enhanced Hsp gene expression in the presence of oxygen. Similarly, the rpsL141 allele, which reduces mistranslation and Hsp gene expression, failed to affect the Hsp regulon in cells grown anaerobically. Increased production of Hsps in response to starvation is associated with increased mistranslation and was demonstrated to likewise require the presence of oxygen. Thus, mistranslation triggered by starvation or mutations in the accuracy centre of the ribosome appear to elevate Hsp gene expression via an oxidative modification of mistranslated proteins. In contrast, Hsp gene induction during temperature upshifts was independent of oxygen availability. The data further suggest that it is the oxidative modification of mistranslated DnaK substrates rather than oxidation of DnaK itself that triggers Hsp gene expression upon starvation. PMID- 16359342 TI - Use of proton pump inhibitor as a diagnostic test in NCCP. AB - Non-cardiac chest pain (NCCP) is the most common extra-esophageal manifestation of gastroesophageal reflux disease. The proton pump inhibitor empirical trial (PPI test) is a simple non-invasive, accurate and cost-saving test for evaluating patients with GERD-related NCCP. The review will discuss the use of the PPI test as a diagnostic test for NCCP. PMID- 16359343 TI - Treatment of noncardiac chest pain. PMID- 16359344 TI - How do we define non-cardiac chest pain? AB - Noncardiac chest pain (NCCP) is a heterogeneous disorder associated with substantial health-care costs and resource utilization. NCCP is defined by recurrent episodes of substernal chest pain in patients lacking a cardiac cause after a comprehensive evaluation. The magnitude of the problem is quite high because of fear of serious or life-threatening heart diseases. Patients with chest pain who present for the first time to ambulatory care or to the emergency room, only 11% to 39% are ultimately diagnosed with coronary artery disease. The likely causes of NCCP are numerous and often overlap. Diagnosing NCCP is difficult because NCCP remains a diagnosis of exclusion that encompasses heterogeneous patient populations. First, cardiac ischemic must be excluded and coronary angiography remains the gold standard. Once cardiac causes have been ruled out, a diagnosis of NCCP is made. Mostly, the source for NCCP originates in essence from the esophagus. Gastroesophageal reflux disease (GERD) is the most common esophageal diseases present in patients with NCCP. An initial empiric trail of high-dose acid suppression is the most cost-effective measure in the management of these patients. When a diagnostic workup is chosen, it centers on upper gastrointestinal endoscopy, 24-hr esophageal pH monitoring and esophageal manometry. PMID- 16359345 TI - Rabeprazole experience in Korea; focus on CYP2C19 polymorphism. PMID- 16359346 TI - CYP2C19 genotype and the PPIs--focus on rabeprazole. AB - Amongst all the proton pump inhibitors (PPI), the hepatic metabolism of rabeprazole is least dependent on the CYP4502C19 system. Rabeprazole is therefore the PPI least affected by CYP4502C19 genetic polymorphism. This unique feature of rabeprazole complements rabeprazole's fast onset of action, and may lead to profound and consistent inhibition of gastric acid secretion in the treatment of acid-related disorders. PMID- 16359347 TI - The pathophysiology of non-cardiac chest pain. AB - Various underlying mechanisms have been described in patients with non-cardiac chest pain (NCCP). By far, gastroesophageal reflux disease (GERD) is the most common cause and thus requires initial attention when patients with NCCP are managed. Esophageal dysmotility can be demonstrated in 30% of the NCCP patients, but appears to play a very limited role in symptom generation. A significant number of patients with NCCP lack any evidence of GERD and have been consistently shown to have reduced perception thresholds for pain. Peripheral and/or central sensitization have been suggested to be responsible for visceral hypersensivity in NCCP patients. Further understanding of the underlying mechanisms for pain in patients with NCCP will likely improve our current therapeutic approach. PMID- 16359348 TI - Being hurt and hurting others: children's narrative accounts and moral judgments of their own interpersonal conflicts. AB - Children's narrative accounts and moral evaluations of their own interpersonal conflicts with peers were examined. Girls and boys (N = 112) in preschool (M= 4.8 years), first grade (M = 6.9 years), fifth grade (M = 10.9 years), and tenth grade (M = 16.2 years) provided one narrative of a time when they had been hurt by a peer ("victim"), and one of a time when they had hurt a peer ("perpetrator"). Victim and perpetrator narratives were equally long and detailed and depicted similar types of harmful behaviors, but differed significantly in terms of various measures of content and coherence. Narratives given from the victim's perspective featured a self-referential focus and a fairly coherent structure. When the same children gave accounts of situations in which they had been the perpetrators, their construals were less coherent and included multiple shifts between references to their own experience and the experience of the other. Children's moral judgments also varied by perspective, with the majority of victims making negative judgments and nearly half the perpetrators making positive or mixed judgments. These differences in moral judgments were related to the distinct ways in which victims and perpetrators construed conflict situations. Age differences were also found in both narrative construals and moral evaluations, but regardless of their age children construed conflict situations differently from the victim's and the perpetrator's perspectives. By integrating, within the study of moral development, children's interpretations of the social interactions that are at the basis of moral thinking, this approach brings us a step closer to conceptualizing the study of children's moral behavior. PMID- 16359349 TI - Human agency and the "joints" of social experience: a commentary on Wainryb, Brehl, and Matwin. PMID- 16359350 TI - Paediatric transfusion. AB - The requirements of children undergoing transfusion should be considered as a distinct entity from those of adults. Neonates are particularly vulnerable and there have been concerns over infective or toxic risks to this group. Neonates may also have more acute side-effects as a result of their small blood volume. Most children who are transfused will have a good life expectancy, so long-term side-effects will be more significant than for adults. In Britain, there are a number of transfusion components with neonatal specifications, but there appears to be some confusion, among both medical and laboratory staff, as to the appropriate use of these. Although there are many paediatric guidelines on the use of blood, there is a lack of evidence underlying these. However, there are trends to decreased blood usage in neonates and ongoing studies to investigate the appropriate use of blood for children. PMID- 16359351 TI - The relevance of natural killer cell human leucocyte antigen epitopes and killer cell immunoglobulin-like receptors in bone marrow transplantation. AB - The discovery that killer cell immunoglobulin-like receptors (KIR) interact with genetically polymorphic epitopes on class I human leucocyte antigen (HLA) molecules and that the KIR receptor repertoire itself is genetically variable has led to investigation of the relevance of the KIR system to stem cell transplantation. A number of retrospective studies of transplant outcome have now demonstrated either beneficial or deleterious effects of mismatching for class I natural killer (NK) epitopes. A smaller number of studies have shown effects of the donor and/or patient KIR repertoire on outcome, irrespective of the patient and donor HLA type. The most parsimonious interpretation of the data, which are often conflicting, is that the effect of NK epitope matching is very much dependent on transplant protocols, with the extent of donor T-cell depletion possibly being the most important variable. A clearer picture of the role of matching for NK epitopes and the KIR-receptor repertoire of the donor is needed. PMID- 16359352 TI - Viral safety of Nanogam, a new 15 nm-filtered liquid immunoglobulin product. AB - BACKGROUND AND OBJECTIVES: Producers of plasma derivatives continuously improve the viral safety of their products by, for example, introducing additional virus reducing steps into the manufacturing process. Here we present virus-elimination studies undertaken for a number of steps employed in a new manufacturing process for liquid intravenous immunoglobulin (Nanogam) that comprises two specific virus reducing steps: a 15-nm filtration step combined with pepsin treatment at pH 4.4 (pH 4.4/15NF); and solvent-detergent (SD) treatment. The manufacturing process also includes precipitation of Cohn fraction III and viral neutralization, which contribute to the total virus-reducing capacity of the manufacturing process. In addition, the mechanism and robustness of the virus-reducing steps were studied. MATERIALS AND METHODS: Selected process steps were studied with spiking experiments using a range of lipid enveloped (LE) and non-lipid-enveloped (NLE) viruses. The LE viruses used were bovine viral diarrhoea virus (BVDV), human immunodeficiency virus (HIV) and pseudorabies virus (PRV); the NLE viruses used were parvovirus B19 (B19), canine parvovirus (CPV) and encephalomyocarditis virus (EMC). After spiking, samples were collected and tested for residual infectivity, and the reduction factors were calculated. For B19, however, removal of B19 DNA was measured, not residual infectivity. To reveal the contribution of viral neutralization, bovine parvovirus (BPV) and hepatitis A virus (HAV) were used. RESULTS: For the pH 4.4/15NF step, complete reduction (> 6 log(10)) was demonstrated for all viruses, including B19, but not for CPV (> 3.4 but < or = 4.2 log(10)). Robustness studies of the pH 4.4/15NF step with CPV showed that pH was the dominant process parameter. SD treatment for 10 min resulted in complete inactivation (> 6 log(10)) of all LE viruses tested. Precipitation of Cohn fraction III resulted in the significant removal (3-4 log(10)) of both LE and NLE viruses. Virus-neutralization assays of final product revealed significant reduction (> or = 3 log(10)) of both BPV and HAV. CONCLUSIONS: The manufacturing process of Nanogam comprises two effective steps for the reduction of LE viruses and one for NLE viruses. In addition, the precipitation of Cohn fraction III and the presence of neutralizing antibodies contribute to the total virus-reducing capacity of Nanogam. The overall virus-reducing capacity was > 15 log(10) for LE viruses. For the NLE viruses B19, CPV and EMC, the overall virus-reducing capacities were > 10, > 7 and > 9 log(10), respectively. Including the contribution of immune neutralization, the overall virus-reducing capacity for B19 and HAV is estimated to be > 10 log(10). PMID- 16359353 TI - Factors influencing factor VIII activity in frozen plasma. AB - BACKGROUNDS AND OBJECTIVES: Fresh frozen human plasma is an important raw material in the production of coagulation factor concentrates used in patients with haemorrhagic disorders. The aim of the study was to determine how the handling of plasma influences the recovery of coagulation factor VIII activity (FVIII:C), i.e. the influence of time between donation and freezing, of the freezing time and of the ice front velocity. We also studied a tentative eutectic point in human plasma. MATERIALS AND METHODS: Aliquots of plasma from 12 different donors were kept at room temperature for 2, 4 and 6 h before start of freezing. We achieved fast freezing with a freezer that blows cooled air at a high velocity on the plasma containers. Freezing times were 0.5, 1, 4 and 24 h. Temperature was registered continuously during freezing. Plasma and NaCl solutions were frozen slowly to investigate the eutectic point. RESULTS: Storage at room temperature for 6 h caused a small but statistically significant decrease in FVIII:C. Slow freezing with programmed freezing times of 4 and 24 h caused a more pronounced drop in FVIII:C as compared to that of 30 and 60 min. We found no eutectic point in plasma or in plasma with addition of 2 % (w/v) NaCl. CONCLUSION: For an optimal yield of FVIII, freezing should start within 4 h after plasma donation. We propose the use of the term 'ice front velocity' instead of 'freezing speed', taking into consideration that the volume and shape of plasma containers may differ. We found only a marginal loss of FVIII:C when the ice front velocity was 26 mm/h or faster, but a significant loss when it was 9 mm/h or slower. We recommend freezing times of 60 min or shorter. We were not able to demonstrate any eutectic point in human plasma. We therefore recommend that the term eutectic point should not be used as a reference temperature in guidelines on plasma handling. PMID- 16359354 TI - Up to 21-day banked red blood cells collected by apheresis and stored for 14 days after automated wash at different times of storage. AB - BACKGROUND AND OBJECTIVES: A closed-system technology (ACP-215, Haemonetics, Braintree, MA) enables automated washing and extended storage of frozen red blood cells (RBC). This technology was applied to wash banked RBC for removal of undesirable protein and metabolites before transfusion. We studied protein and metabolite depletion as well as RBC metabolism and viability up to 14 days postwash with regard to various pre-storage times. MATERIALS AND METHODS: Thirty RBC units were collected by means of apheresis and subdivided into three arms based on prewash storage time period (6 days/group 1, 14 days/group 2, 21 days/group 3). Wash efficacy (protein depletion, IgA), RBC metabolism (pH, lactate, potassium, haemolysis) and cell viability (ATP) were analysed immediately and 14 days after washing. RESULTS: Total protein and IgA postwash were lowered by automated wash in all groups and uniformly met EC guidelines. Potassium (mmol/l) was below 1.2 mmol/l postwash and significantly below prewash values in all groups, even after 14 days of storage (prewash vs. postwash; P < 0.05). RBCs washed after 14 and 21 days, respectively, showed significantly lower pH values and lower ATP content than RBCs washed after only 6 days of storage. Haemolysis rate remained significantly below 0.8%, the maximum level recommended by the EC guidelines, immediately and 14 days after washing in all units. CONCLUSION: Our data confirm that RBC units banked up to 21 days can be effectively protein- and potassium-depleted with the ACP-215 independent from prewash storage time. With respect to high ATP levels and pH, postwash storage of 2 weeks should be limited to units not older than 7 days before wash. This new washing technology ensures better standardization in washed RBC and provides blood centres with a logistical alternative to 24-h washed RBC products. PMID- 16359355 TI - Recombinant activated factor VII (rFVIIa/NovoSeven) in intractable haemorrhage: use of a clinical scoring system to predict outcome. AB - BACKGROUND AND OBJECTIVES: Recombinant activated factor VII (rFVIIa/NovoSeven) has been advocated in the treatment of life-threatening haemorrhage, but appropriate clinical indications remain uncertain. The aim of this study was to detect factors predictive of outcome and to incorporate them into a prognostically significant scoring system. MATERIALS AND METHODS: Thirty-six patients received rFVIIa for uncontrolled surgical, traumatic or obstetric bleeding in the Northern Region of the UK over a 45-month period. Clinical, laboratory and outcome data were examined. Characteristics of survivor and non survivor groups were compared. A prognostic scoring system was evaluated retrospectively according to the presence of coagulopathy, renal impairment, hypothermia, greater than 10 units of red cell transfusion, advanced age and obstetric indication, with patients allocated to low, intermediate and high-risk groups. RESULTS: Clinical response occurred in 26 patients (72%) with a reduction in prothrombin time and blood product requirements. Death occurred in 19 (53%). Four patients (11%) suffered thrombotic events. Survivors were younger than non survivors and less likely to have coagulopathy, renal impairment or hypothermia at the time of administration. Survivors were more likely to have had an initial clinical response in terms of an immediate reduction in haemorrhage. Non survivors were transfused a greater number of red cell units prior to administration. Survival varied according to prognostic score; low-risk patients had a survival rate of 85%, intermediate-risk patients had a survival rate of 50% and high-risk patients had a survival rate of 18%. CONCLUSIONS: FVIIa has a role in the cessation of haemorrhage, but may not improve survival. Use of a clinical scoring system may help to predict outcome. PMID- 16359356 TI - Comparison of flow cytometry vs. a haematology cell analyser-based method to guide the optimal time-point for peripheral blood stem cell apheresis. AB - BACKGROUND AND OBJECTIVES: For timing the onset of apheresis, parameters obtained by flow cytometry and by a haematological cell analyser were compared. MATERIALS AND METHODS: Haematopoietic cell counts (n = 159) were performed by two different methods; CD34 analyses by flow cytometry, immature myeloid information (IMI) and human progenitor cell counts (HPC) by a haematological cell analyser. RESULTS: Comparing the IMI total results with CD34+ analyses (n = 159) revealed a correlation of r = 0.46 (P < 0.05). Similar results were obtained for HPC (r = 0.44; P < 0.05). CONCLUSION: The haematology analyser-based method does not allow the precise determination of absolute haematopoietic stem cell numbers and is thus not able to replace flow cytometry for the monitoring of peripheral blood stem cell counts. PMID- 16359357 TI - HIV-1 NAT minipool during the pre-seroconversion window period: detection of a repeat blood donor. AB - BACKGROUND: The introduction of nucleic acid amplification technology (NAT) for screening pooled or individual donations remarkably improved the safety of blood products. The size of mini-pooled NAT is considered critical for identification of HIV-1 infected donors during preseroconversion phase of infection. We describe a case of HIV-1 infection in a serologically negative repeat blood donor identified by 16 minipool (MP) NAT. MATERIALS AND METHODS: The donation was tested by Roche Cobas AmpliScreen HIV-1 Test with manual extraction (MultiPrep Specimen Processing Procedure). The sensitivity of different MP sizes was observed. Serial samples of infected donor were examined with different third and fourth generation HIV-1 serological assays. RESULTS: In the index donation viral load was 515 copies/ml corresponding to about 50 IU when diluted in 16 MP. Abbott third and fourth generation EIA tests detected the seroconversion four days later the index donation. CONCLUSION: The report emphasizes the relevance of a very small size of MP to really reduce the window serologic phase of current EIA test by HIV-1 NAT test. PMID- 16359358 TI - Paid vs. unpaid donors. PMID- 16359359 TI - Three-dimensional reconstruction of the angioarchitecture of the ciliary body of the West Indian manatee (Trichechus manatus). AB - OBJECTIVE: To examine the angioarchitecture of the ciliary body in the West Indian manatee (Trichechus manatus), through the use of three-dimensional reconstruction. PROCEDURE: Specimens from West Indian manatee were preserved in 10% buffered formalin, embedded in paraffin, serial sectioned and stained by Masson trichrome for light microscopic three-dimensional reconstruction and evaluation. RESULTS: The network of blood vessels in the ciliary processes of the West Indian manatee is fed by the major arterial circle that lies mostly near the base of the iris. The branching arterioles give rise to a capillary-sinusoidal bed that extends internally along each process, emptying into two sets of veins, one being elevated. The elevated and nonelevated veins join posteriorly before emptying into the choroidal venous system. CONCLUSIONS: The angioarchitecture of the ciliary body of the West Indian manatee is clearly unique when compared to those previously examined in land mammals. Three-dimensional reconstruction of paraffin sections is an effective means to evaluate vascular patterns in ocular specimens, especially those unavailable for corrosion casting. PMID- 16359360 TI - Comparative anatomy of the ciliary body of the West Indian manatee (Trichechus manatus) and selected species. AB - OBJECTIVE: To examine the anatomy of the ciliary body in the West Indian manatee (Trichechus manatus), paying close attention to its vascularization and to compare to those of its distant relative, the African elephant (Loxodonta africana), the amphibious hippopotamus (Hippopotamus amphibius) and the aquatic short-finned pilot whale (Globicephala macrorhynchus). PROCEDURE: Specimens from each species were preserved in 10% buffered formalin, and observed stereomicroscopically before being embedded in paraffin, sectioned and stained by Masson trichrome, hematoxylin and eosin, and periodic acid-Schiff for light microscopic evaluation. RESULTS: The network of blood vessels in the ciliary processes of the West Indian manatee appear to have an intricate pattern, especially with regard to venous outflow. Those of the elephant are slightly less complex, while those of the hippopotamus and whale have different vascular patterns within the ciliary body. Musculature within the ciliary body is absent within the manatee and pilot whale. CONCLUSIONS: In general, there appears to be a direct relationship between the increased development of vasculature and the loss of musculature within the ciliary bodies of the aquatic and amphibious mammals presently studied. Specifically, the ciliary body of the West Indian manatee has a comparatively unique construction, especially with regard to its vasculature. PMID- 16359361 TI - Ocular signs of canine monocytic ehrlichiosis: a retrospective study in dogs from Barcelona, Spain. AB - Canine monocytic ehrlichiosis (CME) is a tick-borne disease caused by the rickettsia Ehrlichia canis. Ocular lesions are a common feature of the disease and can be present in all stages. The purpose of this retrospective study was to determine the prevalence, type and response to treatment of ocular lesions associated with monocytic ehrlichiosis in 46 affected dogs presented to the Autonomous University of Barcelona-Veterinary Teaching Hospital (UAB-VTH) from January 2000 to December 2002. Dogs were included in the study only if they had a positive serologic test for E. canis and information about the clinical outcome was available. Eighteen breeds were represented, with the German Shepherd dog (n = 6) being the most common. There were 25 intact and three castrated males, and 16 intact and two neutered females. Twenty dogs (43.4%) were between 5 and 10 years old. Seventeen dogs (37% of all cases of monocytic ehrlichiosis diagnosed during the study period) had ocular signs, and 11 dogs (64.7% of the ocular cases) had only ocular lesions without apparent systemic signs. Exudative retinal detachment was the most common ocular manifestation; other prevalent findings included anterior exudative uveitis and optic neuritis. Five of the 17 cases with ocular lesions (29.4%) had ocular bleeding disorders (hyphema or retinal hemorrhages). All the dogs with ocular disease presented with bilateral signs. Dogs with posterior segment disease had titers against E. canis that were > or = 1 : 320, while lower titers were noted in dogs with anterior exudative uveitis. Two dogs presented with chronic autoimmune panuveitis after ehrlichiosis treatment. Canine ehrlichiosis should be considered in the differential diagnosis of exudative retinal detachment and anterior uveal inflammatory lesions. PMID- 16359362 TI - Major histocompatibility class II expression in the normal canine cornea and in canine chronic superficial keratitis. AB - OBJECTIVE: To compare the expression of major histocompatibility complex (MHC) class II antigen in the corneas of normal dogs and dogs affected with chronic superficial keratitis (CSK). METHODS: MHC class II expression was determined in frozen sections of normal canine cornea and cornea from lesions of CSK by immunohistochemistry using a monoclonal antibody directed against the canine MHC class II molecule. Langerhans cell phenotype was determined morphologically and by histochemical determination of ATPase activity. To determine the influence of gamma interferon on expression of MHC class II molecules by corneal cells, corneal explants were cultured with the cytokine and MHC class II expression determined as above. RESULTS: Numerous MHC class II-expressing cells were demonstrated within the stroma and epithelium of the normal corneal limbus and conjunctival epithelium while very little MHC class II expression was detected in the central region of normal canine cornea. In limbal and conjunctival epithelium, cells expressing MHC class II antigen showed ATPase activity, suggesting that they were Langerhans cells. Corneas from dogs with CSK showed MHC class II expression associated with stromal cells, some of which exhibited a dendritic morphology while most were lymphocytic. Corneal epithelial cells within the lesion also aberrantly expressed MHC class II. Corneal explants expressed MHC class II to varying degrees after differing periods of incubation with the cytokine gamma interferon. CONCLUSIONS: While the normal central cornea has little MHC class II expression, aberrant expression occurs in CSK, associated with secretion of gamma interferon by infiltrating CD4-expressing lymphocytes. Although this change is likely to be a secondary feature of the CSK lesion, increased MHC class II expression may play a part in perpetuating the corneal inflammation seen in the disease. PMID- 16359363 TI - Conjunctival impression cytology in dogs. AB - OBJECTIVE: Ocular conjunctivas of healthy dogs were studied by conjunctival impression cytology for evaluation of feasibility, protocol standardization, and normal cytologic pattern recognition of this technique. ANIMALS STUDIED: Twenty healthy, adult, cross-breed dogs. PROCEDURES: Samples of the bulbar conjunctiva were collected after instillation of topical anesthetic drops at the ocular surface. Impression cytology was performed by applying asymmetric strips of Millipore filter on the superior temporal bulbar conjunctiva near the limbus. The filter strip was gently pressed against the conjunctiva for 5 s and removed with a peeling motion. Samples were immediately fixed in 95% ethyl alcohol, stained with periodic acid-Schiff and hematoxylin, and mounted on slides cover-slipped using synthetic resin. The slides were examined by light microscopy. RESULTS: Microscopic examination of the impressions revealed superficial, intermediate and basal epithelial cells arranged in sheets. Keratinized epithelial cells, goblet cells and leukocytes, as well as cellular debris and mucus were observed. CONCLUSIONS: Feasibility of impression cytology for sampling the bulbar conjunctiva of the dog and the standardization the the proposed protocol was shown. The results allowed the recognition the the normal cytologic pattern of healthy conjunctivas in dogs. PMID- 16359364 TI - Detection of cone dysfunction induced by digoxin in dogs by multicolor electroretinography. AB - It is difficult to detect discrete cone function with the present conventional electroretinography (ERG) examination. In this study, we developed contact electrodes with a built-in color (red (644 nm), green (525 nm), or blue (470 nm)) light source (color LED-electrode), and evaluated an experimental model of digoxin in the dog. First, 17 normal Beagle dogs were used to determine which electrode works well for color ERG measurement on dogs. Then, color ERG was performed on seven normal Beagle dogs at various points during a 14-day period of digoxin administration. A single daily dose of 0.0125 mg/kg/day, which is within the recommended oral maintenance dosage range for dogs, was administered orally for 2 weeks. Ophthalmic examination, measurement of plasma concentration of digoxin, and color ERG examination were performed. On first examination, amplitudes of all responses were significantly (P < 0.01) lower with the red, than with the blue and green electrodes during ERG recording. In ERG using the red electrode, the standard deviation was large. According to these preliminary results, the red electrode was not used in the experimental dog model with digoxin. In the digoxin administrated animals, no significant change was observed in the ophthalmic examination findings. The digoxin level increased steadily throughout the dosing period but was always within the therapeutic range for dogs. In rod ERG, no abnormalities were detected with any electrode. In standard combined ERG, decreased amplitude of the a-wave was detected with every electrode. In single flash cone ERG, prolongation of implicit time was detected by color ERG with the blue and green electrodes. In 30-Hz flicker ERG, decreased amplitude was detected only by color ERG with the blue electrode. The decreased amplitude and prolonged implicit time recovered after termination of digoxin administration. Cone dysfunction induced by digoxin in the dog was revealed by multicolor ERG using blue and green LED-electrodes. Multi-color ERG was useful for detecting cone type-specific dysfunction in the dog. PMID- 16359365 TI - Congenital stationary night blindness in a Thoroughbred and a Paso Fino. AB - This report documents congenital stationary night blindness (CSNB) in two non Appaloosa horse breeds (Thoroughbred and Paso Fino). History of vision impairment since birth, normal ocular structures on ophthalmic examination, and electroretinographic findings were consistent with CSNB. In one horse (Thoroughbred), a 9-year follow-up was carried out. In the Paso Fino, severe vision impairment from birth to approximately 1 year of age in both dim and bright light situations led to humane euthanasia and histopathologic confirmation of the disorder. PMID- 16359366 TI - Intracranial meningioma causing internal ophthalmoparesis in a dog. AB - A 10-year-old, spayed female, Irish water spaniel was presented with a 2-week history of anisocoria characterized by mydriasis of the right eye compared to the left eye in ambient light. Ophthalmic and neurological examinations, combined with pharmacological testing, identified a disease process affecting the right parasympathetic nucleus of cranial nerve 3 (CN III) and/or the parasympathetic component of CN III. Magnetic resonance imaging (MRI) identified a mass involving the right midbrain and extending caudally to the rostral border of the medulla oblongata. The dog became comatose within 12 h following MRI and was euthanized. Histopathology identified the intracranial mass as a meningioma. PMID- 16359371 TI - The prevention of infection post-transplant: the role of prophylaxis, preemptive and empiric therapy. AB - The close linkage of infection with the nature and intensity of the transplant immunosuppressive program has led to the concept of the therapeutic prescription. This has two components: an immunosuppressive one to prevent or treat rejection and graft-versus-host disease and an antimicrobial one to make it safe. This review provides a conceptual framework to approach the risk and risk periods for infection in solid organ and hematopoietic stem cell transplant recipients as well as an approach to antimicrobial use in this population. PMID- 16359372 TI - Marked erythrocyte microcytosis under primary immunosuppression with sirolimus. AB - The preliminary observation of marked erythrocyte microcytosis in patients treated with sirolimus (SRL) and mycophenolate mofetil (MMF) has been evaluated as part of a prospective study comparing SRL and cyclosporin A (CsA) as a primary immunosuppressant. Normal risk de novo kidney recipients were randomized either to SRL or to CsA. Additional immunosuppressants consisted of MMF and prednisone. In patients with erythrocyte microcytosis, iron deficiency was excluded by measuring serum ferritin and transferrin saturation rate. Fifty-nine patients (30 in SRL and 29 in CsA) were included. Mean corpuscular volume (MCV) (fl) on day 7 was 91.7 +/- 4.8 in SRL group versus 91.4 +/- 4.2 in CsA group (P = 0.77), whereas mean MCV on day 183 post-transplant was 78.5 +/- 3.8 in SRL group versus 88.4 +/- 3.4 in CsA group (P < 0.0001). Hemoglobin concentration (g/dl) was not significantly different. Only two patients in SRL group presented decreased transferrin saturation rate. Marked erythrocyte microcytosis without persistent anemia was observed in patients treated with SRL and MMF. PMID- 16359373 TI - Effect of donor-specific transfusions on the outcome of renal allografts in the cyclosporine era. AB - Despite the introduction of new immunosuppressive agents, a steady decline of functioning renal allografts after living donation is observed. Thus nonpharmacological strategies to prevent graft loss have to be reconsidered, including donor-specific transfusions (DST). We introduced a cyclosporine-based DST protocol for renal allograft recipients from living-related/unrelated donation. From 1993 to 2003, 200 ml of whole blood, or the respective mononuclear cells from the potential living donor were administered twice to all of our 61 recipient candidates. The transplanted subjects were compared with three groups of patients without DST from the Collaborative Transplant Study (Heidelberg, Germany) during a 6-year period. Six patients were sensitized without delay for a subsequent cadaveric kidney. DST patients had less often treatment for rejection and graft survival was superior compared with subjects from the other Swiss transplant centers (n = 513) or from Western Europe (n = 7024). To diminish the probability that superior results reflect patient selection rather than effects of DST, a 'matched-pair' analysis controlling for relevant factors of transplant outcome was performed. Again, this analysis indicated that recipients with DST had better outcome. Thus, our observation suggests that DST improve the outcome of living kidney transplants even when modern immunosuppressive drugs are prescribed. PMID- 16359374 TI - Thrombotic and nonthrombotic hepatic artery complications in adults and children following primary liver transplantation with long-term follow-up in 1000 consecutive patients. AB - Arterial complications have a major impact on survival after liver transplantation (LTx). The aim of this study was to examine arterial complications in adults and children after LTx. A total of 1000 consecutive primary LTx patients [mean age 40.5 years: 600 males, 400 females, 834 adults; 166 children (age <18 years)] were studied. Forty-two patients (4.2%; 31 adults, 11 children) developed hepatic artery thrombosis (HAT). Thrombosis in children occurred significantly early (mean 5.4 days) compared with adults (mean 418.7 days, P = 0.0001). Nonthrombotic complications occurred in 30 patients (29 adults, one child). Overall, 13-year patient survival after HAT was 43.2% (72.7% children, 32.9% adults). For nonthrombotic complications, 54.3% of adults died and 69.4% grafts were lost. An overall incidence of 4.2% thrombotic and 3.2% nonthrombotic complications was observed. Rate of HAT was higher in children, but survival was better compared with adults. PMID- 16359375 TI - Use of anti-HBc positive allografts in adult liver transplantation: toward a safer way to expand the donor pool. AB - The use of livers from anti-hepatitis B core (HBc) positive donors can alleviate donor shortage. Nineteen of 367 (6%) adults receiving anti-HBc positive allografts [three were hepatitis B antigen (HBsAg) negative, hepatitis B antibody (HBsAb) positive; four were HBsAg positive and 12 were not exposed to hepatitis B viral (HBV) infection] were retrospectively reviewed. In HBsAg negative recipients, immunoprophylaxis (IP) was guided by viral serology and immunohistochemistry (IH) of day 0 and day 7 liver biopsies. If IH was negative, IP was stopped. None of three HBsAg negative, HBsAb positive recipients infected; one (replicating) of four HBsAg positive recipients reinfected and seven of eight (87.5%) HBsAg, HBsAb negative recipients, who did not receive long-term IP, infected after a median time of 2 years (range 1-5); one patient died of liver failure. Four HBsAg, HBsAb negative recipients, receiving life-long IP, remained infection free. Anti-HBc positive donor livers must be directed selectively first to HBsAg positive recipients, next to recipients having HBV antibodies and finally to HBV-naive recipients. Identification of both donor and recipient risk factors for HBV infection before transplantation allows indiscriminate use of antiviral prophylaxis. The necessity for IP therapy should be guided by HBV-DNA testing of donor liver tissue and serum. IH of early liver biopsies is an unreliable marker for predicting antiviral treatment requirements. PMID- 16359376 TI - Mycophenolic acid clinical pharmacokinetics influenced by a cyclosporine C2 based immunosuppressive regimen in renal allograft recipients. AB - Therapeutic drug monitoring of mycophenolic acid (MPA) in combination with cyclosporine 2-h concentration (CsA C2, n = 68) or tacrolimus trough concentration (n = 10) was investigated by repeated measurements of MPA and MPA glucuronide (MPAG) trough concentrations in renal allograft recipients during the first 3 months post-transplant. The acute rejection rate was lower (19% vs. 43%; P < 0.05) in patients achieving CsA C2 target range during the first week (1600 2000 microg/l), n = 26, compared with those who did not, n = 42. Median MPA concentration was 0.9 and 1.2 microg/ml in patients within or below C2 range, respectively (P = 0.19). CsA C2 correlated with MPAG-to-MPA ratio (P < 0.01, r = 0.91) and gamma-glutamyl-transpeptidase (GGT, P < 0.01, r = 0.86). Total MPA concentration increased during the 3 months, but not in patients on tacrolimus. High CsA C2 lowered the acute rejection rate and plasma MPA. High CsA C2 is associated with elevated GGT, probably because of cholestatic effects, which explain the increased MPAG-to-MPA ratio. Increasing MPA concentration is ascribed to per-protocol CsA C2 reductions. In conclusion, CsA may confound the relationship between MPA and the incidence of rejection, and contribute to the difficulty of obtaining a therapeutic range for MPA in clinical practice. PMID- 16359377 TI - Waiting for a thoracic transplant in Eurotransplant. AB - The prospects of patients on the thoracic waiting list are governed by the chance of receiving an organ in time and by the outcome of the transplantation. The former probability is determined by a triad of disease severity, resource size and allocation rules. The aim of this study was to provide an objective description of the distributional effects of the thoracic allocation system in Eurotransplant. It appears that the interpretation of waiting-list outflow indicators is not straightforward and that it is difficult to assess the fairness of an organ allocation system in the framework of changing donor-organ availability. The timing of listing for heart transplantation can substantially be improved; whether this is also true for lung transplantation cannot be determined from the available data. Allocation schemes cannot solve the problem of organ shortage; a shift of attention toward collaboration with procurement professionals is needed. PMID- 16359378 TI - Transplantation of adult recipients by single cadaveric kidneys from pediatric donors weighing < or = 25 kg can be a reliable option. AB - The evidence in favor of transplanting single allografts from cadaveric pediatric donors into adult recipients is equivocal. This study was performed to assess the outcome of transplantation of single kidneys from pediatric donors weighing < 25 kg. Thirty-five adults transplanted by renal allografts from pediatric donors weighing < 25 kg were compared with 30 matched recipients of kidneys from adult donors. Donors in study group were aged 4.2 +/- 2.1 years weighing 16.0 +/- 5.3 kg. In the study group, surgical complications occurred in five of 35 patients, in the control group four of 30. Serum creatinine reached nadir in 47.5 days in study group versus 30 days in controls (P < 0.01). Serum creatinine at 1 and 3 years were comparable in both groups. A 38.9% had proteinuria at 1 year in the study group compared with 22.7% in controls (P = 0.36). One-year graft survival was 91.7% in the study group versus 92.8% for controls. The surgical complications and graft survival in the study group was comparable with that of controls. The incidence of proteinuria may be more frequent, but does not appear to impact graft function. The use of single, as compared with paired, pediatric donor kidneys would allow more patients to be transplanted with equivalent results. PMID- 16359379 TI - Up-regulation of cell cycle regulatory genes after renal ischemia/reperfusion: differential expression of p16(INK4a), p21(WAF1/CIP1) and p27(Kip1) cyclin dependent kinase inhibitor genes depending on reperfusion time. AB - The aim of this study was to evaluate the influence of renal ischemia, cold preservation and reperfusion on the degree of renal kidney senescence. An experimental model of ex vivo renal hemoperfusion was used. Expression of p16(INK4a), p21(WAF1/CIP1) and p27(Kip1) cyclin-dependent kinase inhibitor genes (CDKIGs) was studied immunohistochemically in kidney biopsy samples at baseline and different time points after reperfusion. All three markers were up-regulated in kidney tissue after the reperfusion; however, their activation in different renal cells varied according to the reperfusion time. Expression of p16 was significantly increased in tubular cells at 180 min of reperfusion when compared with the baseline. Activation of p27 was detected in glomerular cells at 15 min and was significantly higher at 60, 120 and 180 min of reperfusion. The marker started increasing in tubular cells at 15 min and was elevated at every time point afterwards. p21 was significantly over-expressed in all renal cells after the reperfusion. It has been shown by the results of the current study that renal ischemia/reperfusion is associated with over-expression of CDKIGs indicating on substantial DNA damage and/or accelerated tissue senescence. For the first time it has been shown that tissue expression of CDKIGs is positively related with the reperfusion time. PMID- 16359380 TI - Toxoplasmosis-associated hemophagocytic syndrome in renal transplantation. AB - Toxoplasmosis is an infrequent, often difficult to diagnose and potentially lethal disease in kidney transplant recipients. Among reported cases, a few were associated with hemophagocytic syndrome (HPS), a rare condition characterized by widespread proliferation of macrophages phagocytizing blood elements, accompanied by fever and pancytopenia. We report here the case of a patient who received a Toxoplasma gondii positive kidney allograft and developed invasive toxoplasmosis 10 days after surgery, with high fever, skin rash, arthralgias, and renal failure, followed by pneumonia, anemia, thrombocytopenia, liver dysfunction, and encephalitis. Mislead by the absence of Toxoplasma on blood smears, alveolar fluid, renal graft biopsy, and negative brain computed tomography, confusion with serum sickness, and simultaneous herpetic infection, we failed to make the right diagnosis and the patient died with septic shock 11 days later. An HPS was revealed by a late bone marrow analysis. This may well be the fourth case ever reported of toxoplasmosis-associated HPS in renal transplant recipients. PMID- 16359381 TI - The use of a composite graft of iliac artery and inferior mesenteric vein in liver transplantation. AB - Innovation may be required for satisfactory arterial reconstruction in liver transplantation, particularly when the vessels obtained from the donor are inadequate. We have used a composite graft of donor iliac artery and recipient inferior mesenteric vein (IMV) between the infrarenal aorta and donor hepatic artery. Postoperative liver function was satisfactory, with normal daily duplex ultrasound scans for the first 2 weeks. At 4 years follow up, graft function is normal, a duplex ultrasound scan shows normal arterial flow and no dilatation of the composite graft, and a magnetic resonance angiogram reveals no evidence of dilatation or thrombosis of the composite graft. This is one of the few reported cases in which a composite graft has been used to arterialize the allograft in liver transplantation. A composite graft of iliac artery and IMV provided a satisfactory outcome in this patient and may be a valuable addition to the arterial grafts available to the liver transplant surgeon. PMID- 16359382 TI - Cardiac transplantation without calcineurin inhibitors. Sirolimus as first immunosuppressive line. Based on one case. PMID- 16359383 TI - More is not always better: a case postrenal transplant large volume diuresis, hyponatremia, and postoperative seizure. PMID- 16359384 TI - AtNAC2, a transcription factor downstream of ethylene and auxin signaling pathways, is involved in salt stress response and lateral root development. AB - An NAC-type transcription factor gene AtNAC2 was identified from Arabidopsis thaliana when expression patterns of the genes from a microarray analysis were examined. The AtNAC2 expression was induced by salt stress and this induction was reduced in magnitude in the transgenic Arabidopsis plants overexpressing tobacco ethylene receptor gene NTHK1. AtNAC2 is localized in the nucleus and has transcriptional activation activity. It can form a homodimer in yeast. AtNAC2 was highly expressed in roots and flowers, but less expressed in other organs examined. In addition to the salt induction, the AtNAC2 can also be induced by abscisic acid (ABA), ACC and NAA. The salt induction was enhanced in the ethylene overproducer mutant eto1-1, but suppressed in the ethylene-insensitive mutants etr1-1 and ein2-1, and in the auxin-insensitive mutant tir1-1when compared with that in wild-type plants. However, the salt induction of AtNAC2 was not significantly affected in the ABA-insensitive mutants abi2-1, abi3-1 and abi4-1. These results indicate that the salt response of AtNAC2 requires ethylene signaling and auxin signaling pathways but does not require ABI2, ABI3 and ABI4, intermediates of the ABA signaling pathway. Overexpression of AtNAC2 in transgenic Arabidopsis plants resulted in promotion of lateral root development. AtNAC2 also promoted or inhibited downstream gene expressions. These results indicate that AtNAC2 may be a transcription factor incorporating the environmental and endogenous stimuli into the process of plant lateral root development. PMID- 16359385 TI - Protein- and energy-mediated targeting of chloroplast outer envelope membrane proteins. AB - While the import of nuclear-encoded chloroplast proteins is relatively well studied, the targeting of proteins to the outer membrane of the chloroplast envelope is not. The insertion of most outer membrane proteins (OMP) is generally considered to occur without the utilization of energy or proteinaceous components. Recently, however, proteins have been shown to be involved in the integration of outer envelope protein 14 (OEP14), whose outer membrane insertion was previously thought to be spontaneous. Here we investigate the insertion of two proteins from Physcomitrella patens, PpOEP64-1 and PpOEP64-2 (formerly known as PpToc64-1 and PpToc64-2), into the outer membrane of chloroplasts. The association of PpOEP64-1 with chloroplasts was not affected by chloroplast pre treatments. Its insertion into the membrane was affected, however, demonstrating the importance of measuring insertion specifically in these types of assays. We found that the insertion of PpOEP64-1, PpOEP64-2 and two other OMPs, OEP14 and digalactosyldiacylglycerol synthase 1 (DGD1), was reduced by either nucleotide depletion or proteolysis of the chloroplasts. Integration was also inhibited in the presence of an excess of an imported precursor protein. In addition, OEP14 competed with the insertion of the OEP64s and DGD1. These data demonstrate that the targeting of several OMPs involves proteins present in chloroplasts and requires nucleotides. Together with previous reports, our data suggest that OMPs in general do not insert spontaneously. PMID- 16359386 TI - Enhanced salt tolerance mediated by AtHKT1 transporter-induced Na unloading from xylem vessels to xylem parenchyma cells. AB - AtHKT1 is a sodium (Na+) transporter that functions in mediating tolerance to salt stress. To investigate the membrane targeting of AtHKT1 and its expression at the translational level, antibodies were generated against peptides corresponding to the first pore of AtHKT1. Immunoelectron microscopy studies using anti-AtHKT1 antibodies demonstrate that AtHKT1 is targeted to the plasma membrane in xylem parenchyma cells in leaves. AtHKT1 expression in xylem parenchyma cells was also confirmed by AtHKT1 promoter-GUS reporter gene analyses. Interestingly, AtHKT1 disruption alleles caused large increases in the Na+ content of the xylem sap and conversely reduced the Na+ content of the phloem sap. The athkt1 mutant alleles had a smaller and inverse influence on the potassium (K+) content compared with the Na+ content of the xylem, suggesting that K+ transport may be indirectly affected. The expression of AtHKT1 was modulated not only by the concentrations of Na+ and K+ but also by the osmolality of non-ionic compounds. These findings show that AtHKT1 selectively unloads sodium directly from xylem vessels to xylem parenchyma cells. AtHKT1 mediates osmolality balance between xylem vessels and xylem parenchyma cells under saline conditions. Thus AtHKT1 reduces the sodium content in xylem vessels and leaves, thereby playing a central role in protecting plant leaves from salinity stress. PMID- 16359387 TI - Abscisic acid-activated SNRK2 protein kinases function in the gene-regulation pathway of ABA signal transduction by phosphorylating ABA response element binding factors. AB - The plant hormone abscisic acid (ABA) induces gene expression via the ABA response element (ABRE) present in the promoters of ABA-regulated genes. A group of bZIP proteins have been identified as ABRE-binding factors (ABFs) that activate transcription through this cis element. A rice ABF, TRAB1, has been shown to be activated via ABA-dependent phosphorylation. While a large number of signalling factors have been identified that are involved in stomatal regulation by ABA, relatively less is known about the ABA-signalling pathway that leads to gene expression. We have shown recently that three members of the rice SnRK2 protein kinase family, SAPK8, SAPK9 and SAPK10, are activated by ABA signal as well as by hyperosmotic stress. Here we show that transient overexpression in cultured cell protoplasts of these ABA-activated SnRK2 protein kinases leads to the activation of an ABRE-regulated promoter, suggesting that these kinases are involved in the gene-regulation pathway of ABA signalling. We further show several lines of evidence that these ABA-activated SnRK2 protein kinases directly phosphorylate TRAB1 in response to ABA. Kinetic analysis of SAPK10 activation and TRAB1 phosphorylation indicated that the latter immediately followed the former. TRAB1 was found to be phosphorylated not only in response to ABA, but also in response to hyperosmotic stress, which was interpreted as the consequence of phosphorylation of TRAB1 by hyperosmotically activated SAPKs. Physical interaction between TRAB1 and SAPK10 in vivo was demonstrated by a co immunoprecipitation experiment. Finally, TRAB1 was phosphorylated in vitro by the ABA-activated SnRK2 protein kinases at Ser102, which is phosphorylated in vivo in response to ABA and is critical for the activation function. PMID- 16359388 TI - Anthocyanidin synthase in non-anthocyanin-producing Caryophyllales species. AB - Red colors in flowers are mainly produced by two types of pigments: anthocyanins and betacyanins. Although anthocyanins are widely distributed in higher plants, betacyanins have replaced anthocyanins in the Caryophyllales. There has been no report so far to find anthocyanins and betacyanins existing together within the same plant. This curious phenomenon has been examined from genetic and evolutionary perspectives, however nothing is known at the molecular level about the mutual exclusion of anthocyanins and betacyanins in higher plants. Here, we show that spinach (Spinacia oleracea) and pokeweed (Phytolacca americana), which are both members of the Caryophyllales, have functional anthocyanidin synthases (ANSs). The ability of ANSs of the Caryophyllales to oxidize trans-leucocyanidin to cyanidin is comparable to that of ANSs in anthocyanin-producing plants. Expression profiles reveal that, in spinach, dihydroflavonol 4-reductase (DFR) and ANS are not expressed in most tissues and organs, except seeds, in which ANS may contribute to proanthocyanidin synthesis. One possible explanation for the lack of anthocyanins in the Caryophyllales is the suppression or limited expression of the DFR and ANS. PMID- 16359389 TI - The BME3 (Blue Micropylar End 3) GATA zinc finger transcription factor is a positive regulator of Arabidopsis seed germination. AB - In many plant species, seed dormancy is broken by cold stratification, a pre chilling treatment of fully imbibed seeds. Although the ecological importance of seed response to cold temperature is well appreciated, the mechanisms underlying the physiological changes during cold stratification is unknown. Here we show that the GATA zinc finger protein expressed in Arabidopsis seeds during cold stratification plays a critical role in germination. Characterization of an enhancer-trap population identified multiple lines that exhibited beta glucuronidase (GUS) expression in the micropylar end of the seed (named Blue Micropylar End, BME lines). One of these lines, BME3, had a T-DNA insertion site in the 5' upstream region of a GATA-type zinc finger transcription factor gene (termed BME3-ZF). The BME3-ZF mRNA accumulated in seeds during cold stratification. Characterization of the BME3-ZF promoter indicated that this gene was activated specifically in the embryonic axis, which was still enclosed by the endosperm. The zinc finger gene knockout plants produced seeds exhibiting deeper dormancy, which showed reduced response to cold stratification. The ungerminated knockout seeds exhibited testa rupture, but failed to penetrate the endosperm layer. Application of gibberellic acid (GA3) rescued impaired germination of knockout seeds without cold stratification, indicating that the normal GA signal transduction pathway is present in the knockout mutants. Expression of GA20 oxidase and GA3-oxidase genes was greatly reduced in the knockout seeds, suggesting the potential involvement of the zinc finger protein in GA biosynthesis. These results suggest that the GATA zinc finger protein is a positive regulator of seed germination. PMID- 16359390 TI - Analysis of ABA hypersensitive germination2 revealed the pivotal functions of PARN in stress response in Arabidopsis. AB - Accumulating evidence suggests that mRNA degradation systems are crucial for various biological processes in eukaryotes. Here we provide evidence that an mRNA degradation system is associated with some plant hormones and stress responses in plants. We analysed a novel Arabidopsis abscisic acid (ABA)-hypersensitive mutant, ahg2-1, that showed ABA hypersensitivity not only in germination, but also at later developmental stages, and that displayed pleiotropic phenotypes. We found that ahg2-1 accumulated more endogenous ABA in seeds and mannitol-treated plants than did the wild type. Microarray experiments showed that the expressions of ABA-, salicylic acid- and stress-inducible genes were increased in normally grown ahg2-1 plants, suggesting that the ahg2-1 mutation somehow affects various stress responses as well as ABA responses. Map-based cloning of AHG2 revealed that this gene encodes a poly(A)-specific ribonuclease (AtPARN) that is presumed to function in mRNA degradation. Detailed analysis of the ahg2-1 mutation suggests that the mutation reduces AtPARN production. Interestingly, expression of AtPARN was induced by treatment with ABA, high salinity and osmotic stress. These results suggest that both upregulation and downregulation of gene expression by the mRNA-destabilizing activity of AtPARN are crucial for proper ABA, salicylic acid and stress responses. PMID- 16359391 TI - LEAFY, TERMINAL FLOWER1 and AGAMOUS are functionally conserved but do not regulate terminal flowering and floral determinacy in Impatiens balsamina. AB - In Impatiens balsamina a lack of commitment of the meristem during floral development leads to the continuous requirement for a leaf-derived floral signal. In the absence of this signal the meristem reverts to leaf production. Current models for Arabidopsis state that LEAFY (LFY) is central to the integration of floral signals and regulates flowering partly via interactions with TERMINAL FLOWER1 (TFL1) and AGAMOUS (AG). Here we describe Impatiens homologues of LFY, TFL1 and AG (IbLFY, IbTFL1 and IbAG) that are highly conserved at a sequence level and demonstrate homologous functions when expressed ectopically in transgenic Arabidopsis. We relate the expression patterns of IbTFL1 and IbAG to the control of terminal flowering and floral determinacy in Impatiens. IbTFL1 is involved in controlling the phase of the axillary meristems and is expressed in axillary shoots and axillary meristems which produce inflorescences, but not in axillary flowers. It is not involved in maintaining the terminal meristem in either an inflorescence or indeterminate state. Terminal flowering in Impatiens appears therefore to be controlled by a pathway that uses a different integration system than that regulating the development of axillary flowers and branches. The pattern of ovule production in Impatiens requires the meristem to be maintained after the production of carpels. Consistent with this morphological feature IbAG appears to specify stamen and carpel identity, but is not sufficient to specify meristem determinacy in Impatiens. PMID- 16359392 TI - Single amino acid variation in barley 14-3-3 proteins leads to functional isoform specificity in the regulation of nitrate reductase. AB - The highly conserved family of 14-3-3 proteins function in the regulation of a wide variety of cellular processes. The presence of multiple 14-3-3 isoforms and the diversity of cellular processes regulated by 14-3-3 suggest functional isoform specificity of 14-3-3 isoforms in the regulation of target proteins. Indeed, several studies observed differences in affinity and functionality of 14 3-3 isoforms. However, the structural variation by which isoform specificity is accomplished remains unclear. Because other reports suggest that specificity is found in differential expression and availability of 14-3-3 isoforms, we used the nitrate reductase (NR) model system to analyse the availability and functionality of the three barley 14-3-3 isoforms. We found that 14-3-3C is unavailable in dark harvested barley leaf extract and 14-3-3A is functionally not capable to efficiently inhibit NR activity, leaving 14-3-3B as the only characterized isoform able to regulate NR in barley. Further, using site directed mutagenesis, we identified a single amino acid variation (Gly versus Ser) in loop 8 of the 14 3-3 proteins that plays an important role in the observed isoform specificity. Mutating the Gly residue of 14-3-3A to the alternative residue, as found in 14-3 3B and 14-3-3C, turned it into a potent inhibitor of NR activity. Using surface plasmon resonance, we show that the ability of 14-3-3A and the mutated version to inhibit NR activity correlates well with their binding affinity for the 14-3-3 binding motif in the NR protein, indicating involvement of this residue in ligand discrimination. These results suggest that both the availability of 14-3-3 isoforms as well as binding affinity determine isoform-specific regulation of NR activity. PMID- 16359393 TI - PICKLE acts during germination to repress expression of embryonic traits. AB - PICKLE (PKL) codes for a CHD3 chromatin remodeling factor that plays multiple roles in Arabidopsis growth and development. Previous analysis of the expression of genes that exhibit PKL-dependent regulation suggested that PKL acts during germination to repress expression of embryonic traits. In this study, we examined the expression of PKL protein to investigate when and where PKL acts to regulate development. A PKL:eGFP translational fusion is preferentially localized in the nucleus of cells, consistent with the proposed role for PKL as a chromatin remodeling factor. A steroid-inducible version of PKL [a fusion of PKL to the glucocorticoid receptor (PKL:GR)] was used to examine when PKL acts to repress expression of embryonic traits. We found that activation of PKL:GR during germination was sufficient to repress expression of embryonic traits in the primary roots of pkl seedlings, whereas activation of PKL:GR after germination had little effect. In contrast, we observed that PKL is required continuously after germination to repress expression of PHERES1, a type I MADS box gene that is normally expressed during early embryogenesis in wild-type plants. Thus, PKL acts at multiple points during development to regulate patterns of gene expression in Arabidopsis. PMID- 16359394 TI - PIF1 is regulated by light-mediated degradation through the ubiquitin-26S proteasome pathway to optimize photomorphogenesis of seedlings in Arabidopsis. AB - Light signals perceived by the phytochrome (phy) family of sensory photoreceptors control multiple aspects of plant development. Recently, PIF1, a phy-interacting basic helix-loop-helix (bHLH) transcription factor, has been shown to negatively regulate facets of the photomorphogenesis of seedlings. Moreover, the transcriptional activation activity of PIF1 is reduced in a phy-dependent manner. In this study we use the luciferase (LUC) activity of the LUC-PIF1 fusion protein as an indicator of the stability of PIF1 in various light conditions. We found that the activity of LUC-PIF1 in both transient and stable transgenic lines is rapidly reduced in light, while the LUC-only control is stable under the same conditions, suggesting that PIF1 is degraded in response to light. Fluence-rate response curves indicate that PIF1 degradation is very sensitive to the quality and quantity of light. The half-life of PIF1 is about 16 min under 10 micromol m 2 sec-1 red light. PIF1 reaccumulates in the subsequent dark period after light induced degradation, signifying that PIF1 not only functions in the dark and during the transition from etiolated to de-etiolated growth, but may also function during diurnal cycles. Inhibitors of the 26S proteasome increased the stability of PIF1, indicating that degradation of PIF1 is mediated by the ubiquitin-26S proteasome pathway. Further, de novo protein synthesis is not required for degradation of PIF1, as the presence of cycloheximide does not prevent degradation of PIF1 in the light. Taken together, these results suggest that the light signals perceived by phys induce the degradation of PIF1 and other phy-interacting factors to optimize photomorphogenesis. PMID- 16359395 TI - Identification of a novel protein, CRR7, required for the stabilization of the chloroplast NAD(P)H dehydrogenase complex in Arabidopsis. AB - An Arabidopsis thaliana mutant, crr7 (chlororespiratory reduction), was isolated using chlorophyll fluorescence imaging to detect reduced activity in NAD(P)H dehydrogenase (NDH). The chloroplast NDH complex is considered to have originated from cyanobacteria in which the NDH complex is involved in respiration, photosystem I (PSI) cyclic electron transport and CO2 uptake. In higher plants the NDH complex functions in PSI cyclic electron transport within the chloroplast. Despite exhaustive biochemical approaches, the entire subunit composition of the NDH complex is unclear in both cyanobacteria and chloroplasts. In crr7 accumulation of the NDH complex was specifically impaired. In vivo analysis of electron transport supported the specific loss of the NDH complex in crr7. CRR7 (At5g39210) encodes a protein of 156 amino acids, including a putative plastid target signal, and does not contain any known motifs. In contrast to CRR2 and CRR4, involved in the expression of chloroplast ndh genes, CRR7 is conserved in cyanobacterial genomes. Although CRR7 did not contain any transmembrane domains, it localized to the membrane fraction of the chloroplast. CRR7 was unstable in the crr2-2 mutant background, in which the expression of ndhB was impaired. These results strongly suggest that CRR7 is a novel subunit of the chloroplast NDH complex. PMID- 16359396 TI - A protocol to analyse cellular dynamics during plant development. AB - In vivo microscopy generates images that contain complex information on the dynamic behaviour of three-dimensional (3D) objects. As a result, adapted mathematical and computational tools are required to help in their interpretation. Ideally, a complete software chain to study the dynamics of a complex 3D object should include: (i) the acquisition, (ii) the preprocessing and (iii) segmentation of the images, followed by (iv) a reconstruction in time and space and (v) the final quantitative analysis. Here, we have developed such a protocol to study cell dynamics at the shoot apical meristem in Arabidopsis. The protocol uses serial optical sections made with the confocal microscope. It includes specially designed algorithms to automate the identification of cell lineage and to analyse the quantitative behaviour of the meristem surface. PMID- 16359397 TI - Maize association population: a high-resolution platform for quantitative trait locus dissection. AB - Crop improvement and the dissection of complex genetic traits require germplasm diversity. Although this necessary phenotypic variability exists in diverse maize, most research is conducted using a small subset of inbred lines. An association population of 302 lines is now available--a valuable research tool that captures a large proportion of the alleles in cultivated maize. Provided that appropriate statistical models correcting for population structure are included, this tool can be used in association analyses to provide high resolution evaluation of multiple alleles. This study describes the population structure of the 302 lines, and investigates the relationship between population structure and various measures of phenotypic and breeding value. On average, our estimates of population structure account for 9.3% of phenotypic variation, roughly equivalent to a major quantitative trait locus (QTL), with a high of 35%. Inclusion of population structure in association models is critical to meaningful analyses. This new association population has the potential to identify QTL with small effects, which will aid in dissecting complex traits and in planning future projects to exploit the rich diversity present in maize. PMID- 16359398 TI - Exploration of jasmonate signalling via automated and standardized transient expression assays in tobacco cells. AB - Although sequence information and genome annotation are improving at an impressive pace, functional ontology is still non-existent or rudimentary for most genes. In this regard, transient expression assays are very valuable for identification of short functional segments in particular pathways, because they can be performed rapidly and at a scale unattainable in stably transformed tissues. Vectors were constructed and protocols developed for systematic transient assays in plant protoplasts. To enhance throughput and reproducibility, protoplast treatments were performed entirely by a liquid-handling robot in multiwell plates, including polyethylene glycol/Ca2+ cell transfection with plasmid mixtures, washes and lysis. All transcriptional readouts were measured using a dual firefly/Renilla luciferase assay, in which the former was controlled by a reporter promoter and the latter by the 35S CaMV promoter, which served as internal normalization standard. The automated protocols were suitable for transient assays in protoplasts prepared from cell cultures of Nicotiana tabacum Bright Yellow-2 and Arabidopsis thaliana. They were implemented in a screen to discover potential regulators of genes coding for key enzymes in nicotine biosynthesis. Two novel tobacco transcription factors were found, NtORC1 and NtJAP1, that positively regulate the putrescine N-methyltransferase (PMT) promoter. In addition, combinatorial tests showed that these two factors act synergistically to induce PMT transcriptional activity. The development and use of high-throughput plant transient expression assays are discussed. PMID- 16359399 TI - Review editorial: prevention of tuberculosis in resource-poor countries with increasing access to highly active antiretroviral treatment. AB - The administration of isoniazid (INH) has been proposed, evaluated and implemented to prevent tuberculosis (TB) disease among patients who are infected with the human immunodeficiency virus (HIV). This strategy has been developed in communities where TB is highly endemic and at a time when antiretroviral (ARV) treatment was not, or was rarely available. Although INH prevention programmes were somewhat pushed to the background due to the worldwide advocacy for ARV drugs, prevention of TB remains of paramount importance. The dual HIV-TB infection poses problems, not only for the individual and his/her clinician but also for the programme manager. We review various aspects of TB preventive treatment in countries with a high prevalence of HIV-TB co-infection and limited resources but with increasing access to ARV treatment. PMID- 16359400 TI - High frequency of multidrug-resistant Mycobacterium tuberculosis isolates in Georgetown, Guyana. AB - Emergence of multidrug-resistant (MDR) Mycobacterium tuberculosis isolates constitutes a threat to public health worldwide. This study aimed at acquiring first epidemiological data for Guyana. Thirty-six M. tuberculosis isolates from patients of the Georgetown Chest Clinic were subjected to susceptibility testing on solid agar and in broth media. Resistance to at least one first-line drug was observed in 8 (22.2%, 95% confidence interval 8.3-36.1%) and simultaneous resistance to rifampicin and isoniazid (MDR) in 4 (11.1%, 95% confidence interval 0.6-21.6%) of the 36 isolates. The risk of infection with resistant isolates was significantly related to earlier antituberculosis therapy (P=0.040). These data indicate a high proportion of resistant M. tuberculosis isolates in Guyana and call for the implementation of control strategies based on an improved laboratory diagnosis of TB. PMID- 16359401 TI - A multi-country cluster randomized controlled effectiveness evaluation to accelerate the introduction of Vi polysaccharide typhoid vaccine in developing countries in Asia: rationale and design. AB - Phase-III vaccine efficacy trials typically employ individually randomized designs intended to ensure that measurements of vaccine protective efficacy reflect only direct vaccine effects. As a result, decisions about introducing newly licensed vaccines into public health programmes often fail to consider the substantially greater protection that may occur when a vaccine is deployed in public health programmes, due to the combination of direct plus indirect vaccine protective effects. Vaccine total protection can be better evaluated with cluster randomized trials. Such a design was considered to generate policy relevant data to accelerate the rationale introduction of the licensed typhoid fever Vi polysaccharide (PS) vaccine in Asia by the Diseases of the Most Impoverished (DOMI) typhoid fever programme. The DOMI's programme multi-country study is one of the largest cluster randomized vaccine trials ever mounted in Asia, which includes approximately 200,000 individuals. Its main objective is to determine the effectiveness of a licensed Vi PS vaccine. The rationale and design of this study are discussed. Preliminary results are presented that determined the final planning of the trial before immunization. Important methodological and practical issues regarding vaccine cluster randomized designs are illustrated. PMID- 16359402 TI - Emergence of W135 meningococcal meningitis in Ghana. AB - Neisseria meningitidis serogroup W135, well known for a long time as a cause of isolated cases of meningococcal meningitis, has recently increasingly been associated with disease outbreaks of considerable magnitude. Burkina Faso was hit by W135 epidemics in the dry seasons of 2002-2004, but only four W135 meningitis cases were recorded between February 2003 and March 2004 in adjoining Ghana. This reconfirms previous findings that bottlenecks exist in the spreading of new epidemic N. meningitidis clones within the meningitis belt of sub-Saharan Africa. Of the four Ghanaian W135 meningitis patients one died and three survived, of whom one had profound neurosensory hearing loss and speech impairment. All four disease isolates were sensitive to penicillin G, chloramphenicol, ciprofloxacin and cefotaxime and had the multi-locus sequence type (ST) 11, which is the major ST of the ET-37 clonal complex. Pulsed-field gel electrophoresis (PFGE) profiles of the Ghanaian disease isolates and recent epidemic isolates from Burkina Faso were largely identical. We conducted meningococcal colonization surveys in the home communities of three of the patients and in the Kassena Nankana District located at the border to Burkina Faso. W135 carriage rates ranged between 0% and 17.5%. When three consecutive surveys were conducted in the patient community with the highest carrier rate, persistence of W135 colonization over a period of 1 year was observed. Differences in PFGE profiles of carrier isolates taken at different times in the same patient community were indicative of rapid microevolution of the W135 bacteria, emphasizing the need for innovative fine typing methods to reveal the relationship between W135 isolates. PMID- 16359403 TI - Risk factors for mortality in Paraguayan children with pneumococcal bacterial meningitis. AB - BACKGROUND: Over the last decade Streptococcus pneumoniae has emerged as the most common bacterial pathogen for meningitis in all age groups, beyond the neonatal period. OBJECTIVE: To determine the epidemiological and clinical characteristics; and risk factors for mortality of pneumoccocal meningitis in children in a developing transitional country. MATERIALS AND METHODS: A retrospective study that included patients<15 years of age admitted at the Instituto de Medicina Tropical of Paraguay, from January 1990 until December 2003 with the diagnosis of bacterial meningitis caused by S. pneumoniae. Clinical and laboratory data were collected and analysed in order to identify risk factors associated with morbidity and mortality outcomes of this infection. RESULTS: Seventy-two patients (between the ages of 35 days and 14 years) were identified. Forty-two per cent of patients had seizures prior to or at the time of admission, 36% were admitted in a comatose state, and 19% with shock. Mortality was 33% (24/72), and 18% of the survivors (11/60) developed severe sequelae. Upon admission, the following variables were strongly correlated with mortality: age<12 months (P=0.007), the presence of seizures (P=0.0001) or development of seizures 48 h after admission (P=0.01), a cerebrospinal fluid (CSF) glucose level of <10 mg/dl (P=0.01), CSF albumin>200 mg/dl (P=0.0003), an absolute blood neutrophil count<2000/mm3 (P=0.006) and a haemoglobin value of <9 g/dl (P=0.0001). CONCLUSIONS: This study confirms the high morbidity and mortality associated with S. pneumoniae meningitis in Paraguay. Certain clinical parameters and laboratory findings in blood and CSF at the time of admission could be used as predictors for mortality or severe sequelae among survivors. PMID- 16359404 TI - High acceptability of voluntary counselling and HIV-testing but unacceptable loss to follow up in a prevention of mother-to-child HIV transmission programme in rural Malawi: scaling-up requires a different way of acting. AB - SETTING: Thyolo District Hospital, rural Malawi. OBJECTIVES: In a prevention of mother-to-child HIV transmission (PMTCT) programme, to determine: the acceptability of offering 'opt-out' voluntary counselling and HIV-testing (VCT); the progressive loss to follow up of HIV-positive mothers during the antenatal period, at delivery and to the 6-month postnatal visit; and the proportion of missed deliveries in the district. DESIGN: Cohort study. METHODS: Review of routine antenatal, VCT and PMTCT registers. RESULTS: Of 3136 new antenatal mothers, 2996 [96%, 95% confidence interval (CI): 95-97] were pre-test counselled, 2965 (95%, CI: 94-96) underwent HIV-testing, all of whom were post test counselled. Thirty-one (1%) mothers refused HIV-testing. A total of 646 (22%) individuals were HIV-positive, and were included in the PMTCT programme. Two hundred and eighty-eight (45%) mothers and 222 (34%) babies received nevirapine. The cumulative loss to follow up (n=646) was 358 (55%, CI: 51-59) by the 36-week antenatal visit, 440 (68%, CI: 64-71) by delivery, 450 (70%, CI: 66 73) by the first postnatal visit and 524 (81%, CI: 78-84) by the 6-month postnatal visit. This left just 122 (19%, CI: 16-22) of the initial cohort still in the programme. The great majority (87%) of deliveries occurred at peripheral sites where PMTCT was not available. CONCLUSIONS: In a rural district hospital setting, at least 9 out of every 10 mothers attending antenatal services accepted VCT, of whom approximately one-quarter were HIV-positive and included in the PMTCT programme. The progressive loss to follow up of more than three-quarters of this cohort by the 6-month postnatal visit demands a 'different way of acting' if PMTCT is to be scaled up in our setting. PMID- 16359405 TI - Factors associated with functional limitations and subsequent employment or schooling in Buruli ulcer patients. AB - OBJECTIVES: To evaluate former Buruli ulcer disease (BUD) patients to assess the factors associated with functional limitations and subsequent employment or schooling. METHODS: The previously validated Buruli ulcer functional limitation score (BUFLS) questionnaire and interviews about educational and professional consequences incurred by BUD. RESULTS: Of 638 participants, 362 (57%) had a functional limitation after a median period of almost 4 years after treatment for BUD. A lesion on a joint, older age, female gender, a lesion on a distal part of an extremity and a persistent wound were found to be independent risk factors for stopping work or education. The same risk factors applied to the development of a functional limitation. Both functional limitations and financial difficulties due to BUD disease often led to job loss and school dropout. CONCLUSIONS: Rehabilitation programmes are urgently needed to diminish the suffering from the functional limitations and employment or schooling problems caused by BUD. PMID- 16359406 TI - Field ecology of sylvatic Rhodnius populations (Heteroptera, Triatominae): risk factors for palm tree infestation in western Ecuador. AB - Most Rhodnius species (Triatominae) are primarily associated with palm trees. They maintain enzootic Trypanosoma cruzi transmission and are responsible for human infection (causing Chagas disease) through the Neotropics. Assessing whether individual palm traits (ecological and/or botanical) may increase the risk of palm infestation by triatomines is relevant in areas where bugs invade houses flying from peridomestic palms. We developed a novel fieldwork approach with that objective, and applied it to study infestation by sylvatic Rhodnius ecuadoriensis in 110 tagua palms (Phytelephas aequatorialis). Palm infestation (23% overall) was non-randomly distributed in our sample. Palms located in anthropic landscapes were frequently infested (>27%, n=92), whereas no bugs were collected from palms surveyed within forest remnants (n=18; P=0.01). The presence of abundant decaying vegetable matter (P=0.001) and (to a lesser extent) epiphytic plants (P=0.049) on palm crowns and stems increased the probability of infestation and was positively correlated with the apparent density of bug colonies (R2=0.68). A trend towards higher infestation rates in male palms (34% vs. 18%) could relate to female palm management (removal of infrutescences and vegetable debris) in areas where palm seeds are harvested. An outline of 'risk palm ecotopes' and environmental management-based strategies for the control of peridomestic, palm tree-living vector populations are proposed. PMID- 16359407 TI - Falciparum malaria in the north of Laos: the occurrence and implications of the Plasmodium falciparum chloroquine resistance transporter (pfcrt) gene haplotype SVMNT. AB - OBJECTIVE: The Pfcrt-gene encodes a transmembrane protein located in the Plasmodium falciparum digestive vacuole. Chloroquine resistant (CQR) strains of African and Southeast Asian origin carry the Pfcrt-haplotype (c72-76) CVIET, whereas most South American and Papua New Guinean CQR stains carry the SVMNT haplotype. METHOD: Eighty-eight samples from an area with reported in vivo Chloroquine and in vitro Amodiaquine-resistance were screened for the K76T mutation and their Pfcrt-haplotype (c72-76) using a new SSOP-ELISA. RESULTS: Hundred percent of the analysed samples showed the K76T mutation which is highly associated with in vivo drug failure. This very high rate of a CQR-marker is alarming in an area were CQ is still used as first line drug. The distribution of the three main Pfcrt-haplotypes was as follows: 68% CVIET, 31% SVMNT, 0% CVMNT. CONCLUSIONS: These data show, for the first time, the South American/PNG haplotype (SVMNT) on mainland Southeast Asia. SVMNT-haplotype and others might be associated with a decreased efficacy of Amodiaquine and could therefore be potential markers for of amodiaquine resistance (AQR). If there is a correlation between AQR and the SVMNT-haplotype as suggested, 31% prevalence of a potential resistance marker is cause for concern. PMID- 16359408 TI - Short communication: high prevalence of the cytochrome P450 2C8*2 mutation in Northern Ghana. AB - Recently, Ghana has changed the first-line treatment of uncomplicated malaria from chloroquine to amodiaquine (AQ) plus artesunate. AQ may cause adverse events such as agranulocytosis and hepatoxicity. The pro-drug AQ is transformed by cytochrome P450 CYP2C8 to the active metabolite N-desethylaminodiaquine. Several polymorphic variants of CYP2C8 are known, some with reduced activity. In 200 randomly selected children from Northern Ghana, we determined the allele frequencies of the CYP2C8 variants CYP2C8*1 (wild type), CYP2C8*2, CYP2C8*3, and CYP2C8*4. We did not detect CYP2C8*3 and CYP2C8*4, but CYP2C8*2 showed an allele frequency of 0.1675. AQ metabolism in patients with CYP2C8*2 may be impaired, and with an increase of AQ based treatment the risk of severe adverse events may mount. PMID- 16359409 TI - Can source reduction of mosquito larval habitat reduce malaria transmission in Tigray, Ethiopia? AB - The development of irrigation schemes by dam construction has led to an increased risk of malaria in Tigray, Ethiopia. We carried out a pilot study near a microdam to assess whether environmental management could reduce malaria transmission by Anopheles arabiensis, the main vector in Ethiopia. The study took place in Deba village, close to a dam; Maisheru village, situated 3-4 km away from the dam, acted as a control. Baseline entomological and clinical data were collected in both villages during the first 12 months. Source reduction, involving filling, draining and shading of potential mosquito-breeding habitats was carried out by the community of Deba in the second year and routine surveillance continued in both villages during the second year. Anopheles arabiensis was highly anthropophilic (Human Blood Index=0.73), biting early in the night before people went to bed. The major breeding habitats associated with the dam were areas of seepage at the dam base (28%), leaking irrigation canals (16%), pools that formed along the bed of streams from the dam (13%), and man-made pools (12%). In the pre intervention year, 5.9-7.2 times more adult vectors were found in the dam village compared with the control village. There was a 3.1% higher prevalence of an enlarged spleen in children under 10 years in the dam village than in the control village during the pre-intervention period, but no statistically significant difference was found in the incidence of falciparum malaria between the two villages during the same period. Source reduction was associated with a 49% (95% CI=46.6-50.0) relative reduction in An. arabiensis adults in the dam village compared with the pre-intervention period. There were very few cases of malaria during the intervention period in both villages making it impossible to judge whether malaria incidence had been reduced. These preliminary findings suggest that in areas of low intensity transmission community-led larval control may be a cheap and effective method of controlling malaria. Further, large-scale studies are needed to confirm these findings. PMID- 16359410 TI - Maternal vitamin A supplementation and immunity to malaria in pregnancy in Ghanaian primigravids. AB - BACKGROUND: Vitamin A supplementation is believed to enhance immune responses to infection but few studies have assessed its effects on anti-malarial immunity, especially during pregnancy when women are at increased risk from both vitamin A deficiency and pregnancy-associated malaria. The pathological effects of malaria in pregnancy are believed to be due to the sequestration of parasites in the placenta mediated via binding of variant surface antigens (VSA) expressed on the surface of P. falciparum infected red blood cells to placental chondroitin sulphate A (CSA). METHODS: We conducted a randomized double-blind controlled trial of vitamin A supplementation in 98 primigravid Ghanaian women to investigate the effects of vitamin A supplementation on levels of IgG antibodies binding to VSA of a clinical, P. falciparum placental isolate and to two isolates selected (or not) for adherence to CSA in vitro (anti-VSACSA IgG or anti-VSA IgG). Placental malarial infection was determined by placental blood smear and histology. RESULTS: Vitamin A supplementation was non-significantly associated with a decreased risk of active or chronic-active placental malarial infection compared to past, resolved infection at delivery, as determined by histology (OR=0.42, P=0.13--adjusted for level of education). After adjustment for differences in baseline values, levels of anti-VSACSA IgG to a placental, CSA adherent isolate (EJ-24) but not to two isolates selected for CSA-adhesion in vitro (FCR3CSA and BusuaCSA), were significantly lower in women receiving vitamin A supplementation than in women receiving placebo (P=0.002). There was no apparent effect of vitamin A supplementation to levels of Ab to non-CSA-adherent parasite isolates. CONCLUSIONS: The data suggest that the reduction in the levels of anti-VSACSA antibodies to the known placental malaria isolate may reflect reduced intensity or duration of placental parasitaemia in women receiving vitamin A supplementation. These observations are of potential public health significance and deserve further investigation. PMID- 16359411 TI - Efficacy of thermal fog application of deltacide, a synergized mixture of pyrethroids, against Aedes aegypti, the vector of dengue. AB - We evaluated the efficacy of indoor and peridomestic thermal fog applications of deltacide, a synergized mixture of pyrethroids (S-bioallethrin 0.7% w/v, deltamethrin 0.5% w/v and piperonyl butoxide 8.9% w/v) against adult populations of Aedes aegypti in Chennai, Tamil Nadu, India. We bioassayed adult caged mosquitoes, counted indoor resting and human landing adult mosquitoes and assessed the percentage of potential breeding sites with Aedes larvae. The bioassay mortalities indicated that the knockdown and killing effect was greater when fogging was applied inside houses rather than around them. Peridomestic thermal fogging reduced the resting and biting populations by 76% and 40%, respectively for the 3 days after treatment, whereas indoor fogging suppressed adult populations for 5 days. PMID- 16359412 TI - Prevalence of enzootic simian viruses among urban performance monkeys in Indonesia. AB - Animal reservoirs are the most important sources of emerging infectious diseases that threaten human populations. Global travel and tourism bring ever-increasing numbers of humans into contact with animals, increasing the likelihood of cross species transmission of infectious agents. Non-human primates come into contact with humans in a variety of contexts and may harbor infectious agents with zoonotic potential. We investigated the prevalence of infection with enzootic simian viruses among 20 urban performance monkeys (Macaca fascicularis) in Jakarta, Indonesia. This report documents for the first time evidence of infection with four simian viruses in urban performance monkeys. Simian foamy virus was detected by PCR in 52.9% of the macaques. Antibodies to simian retrovirus were detected in 10.5% of the macaques. Antibodies to Cercopithecine Herpesvirus 1, were detected in 5.3% of the macaques. Similarly, antibodies to simian T-cell lymphotropic virus were detected in 5.3% of the macaques. No evidence of infection with simian immunodeficiency virus was detected in these macaques. These results suggest that urban performance monkeys are a reservoir for enzootic simian viruses known to be capable of infecting humans. PMID- 16359413 TI - Oesophagostomum bifurcum in non-human primates is not a potential reservoir for human infection in Ghana. AB - In northern Togo and Ghana, human infection with the parasitic nematode Oesophagostomum bifurcum is of major health importance. Elsewhere, oesophagostomiasis is considered a zoonotic infection, non-human primates being the natural host. We examined 349 faecal samples of the olive baboon, mona monkey and black and white colobus monkey from two geographically distinct areas in Ghana, outside the region endemic for O. bifurcum in humans. Using both microscopy and species-specific PCR, we found a high prevalence of O. bifurcum (75-99%) in olive baboons and mona monkeys. The majority of the test-positive faecal samples contained large numbers of larvae after copro-culture (>100). No O. bifurcum was detected in the faeces of the black and white colobus monkeys. Observational studies on the behaviour of the non-human primates, focusing on defecation, food consumption and the sharing of habitat with the local human population, indicated favourable conditions for zoonotic transmission. Given that no human infection with O. bifurcum has been reported from either study area, the present findings support the hypothesis that O. bifurcum from humans in the north of Ghana, and O. bifurcum from olive baboons and/or mona monkeys are distinct. PMID- 16359414 TI - Comment on: Lindblade et al. (2005). Evaluation of long-lasting insecticidal nets after 2 years household use. Tropical Medicine & International Health 10, 1141 50. PMID- 16359415 TI - Survey of the implementation of the recommendations in the Health Service Circular 2002/009 'Better Blood Transfusion'. AB - This report describes the results of questionnaire surveys in 2003 and 2004 on the implementation of the recommendations of the Health Service Circular (HSC) 'Better Blood Transfusion' 2002/009 for improving transfusion practice. These followed a similar survey in 2001 to determine the progress with the implementation of recommendations in the previous Health Service Circular (HSC) 'Better Blood Transfusion' 1998/224. There was a disappointing response rate (47%) to the 2003 survey and evidence for incomplete compliance with the action plan. It was repeated in April 2004 with a systematic approach to encouraging returns, and the response rate was 95%. The results indicate progress in the implementation of Better Blood Transfusion between 2001 and 2004 in relation to increases in the proportion of hospitals with Hospital Transfusion Committees, the training of some staff groups, the number of hospitals with transfusion practitioners, the development of protocols for the use of blood and audit activity. However, the results also indicate the need for further progress in the training of some staff groups, particularly nurses and doctors, the development of Hospital Transfusion Teams, the development of protocols for the appropriate use of blood, the provision of information to patients and the use of peri operative cell salvage. This information should be used to plan further local, regional and national initiatives to implement the Better Blood Transfusion action plan and improve transfusion practice. PMID- 16359416 TI - Consenting to blood: what do patients remember? AB - We sought to characterize the consent process for transfusion and determine its impact on patients' knowledge and level of comfort with receiving blood. We identified all adult patients who had received blood transfusion at a tertiary care centre over 3 months. Patients who were discharged each received a survey that assessed their (1) recall of the consent process, (2) recall of information conveyed, (3) assessment of the discussion's understandability and (4) perceived knowledge of as well as comfort level with transfusion as a result of the discussion. Overall, 80% of respondents recalled discussing and signing an informed consent. Information was mostly conveyed by attending physicians (35%) and consent obtained in the patient's hospital room (38%) or the preadmission clinic (19%). Although the majority recalled the consent process, many did not recall the discussion of specific transfusion risks or alternatives to donor blood (88%). Although the majority felt the discussion was at least somewhat understandable (77%), only 35% felt better informed and more comfortable with accepting blood. Despite implementation of written informed consent for transfusion, patients' recollection and understanding of risks and alternatives remain poor. This suggests the need for improving risk communication. PMID- 16359418 TI - Low dose erythropoietin is effective in reducing transfusion requirements following allogeneic HSCT. AB - Blood transfusions are frequently required for several weeks after allogeneic transplantation due to inadequate erythropoiesis and defective erythropoietin production. Because red cell transfusion is not without complications in this setting, we sought to avoid them using recombinant human erythropoietin (rhEpo) therapy. We treated 53 patients following allogeneic transplantation for haematological malignancy, using rhEpo at a dose of 10 000 units subcutaneously twice weekly. The median time of commencement of rhEpo was 61 days post transplant (range 19-465 days), and the median haemoglobin (Hb) concentration was 9.4 g dL(-1) (range 7.0-10.7 g dL(-1)). Thirty patients responded to rhEpo and required no further transfusion with a median rise in Hb after 2 weeks of therapy of 1.5 g (0.7-4.1 g dL(-1)). Erythropoietin (Epo) was discontinued at a median of 5 weeks (range 2-36), when the median Hb concentration was 12.3 g dL(-1) (range 10.0-14.3 g dL(-1)). Those patients who failed to respond to rhEpo frequently had additional reasons for anaemia including cytomegalovirus (CMV) reactivation and treatment, major ABO incompatibility, disease relapse, graft rejection or other transplant-related complications. We conclude that a short course of rhEpo is an effective treatment for anaemia arising following allogeneic hematopoietic cell transplantation, and can avoid the need for transfusion in this setting. PMID- 16359417 TI - Transfusions of CPDA-1 red blood cells stored for up to 28 days decrease donor exposures in very low-birth-weight premature infants. AB - The goal of this research was to study the safety and the efficacy of transfusing citrate-phosphate-adenine anticoagulant-preservative (CPDA-1) RBC stored for up to 28 days to reduce donor exposures in premature infants. A prospective randomized two-group study was conducted with very low-birth-weight premature infants that received at least one RBC transfusion during hospital stay. Neonates randomly assigned to Group 1 (26 infants) were transfused with CPDA-1 RBC stored for up to 28 days; those assigned to Group 2 (26 infants) received CPDA-1 RBC stored for up to 3 days. Demographic and transfusion-related data were collected. Neonates from both groups showed similar demographics and clinical characteristics. The number of transfusions per infant transfused was 4.4 +/- 4.0 in Group 1 and 4.2 +/- 3.1 in Group 2, and the number of donors per infant transfused was 1.5 +/- 0.8 (Group 1) and 4.3 +/- 3.4 (Group 2), P < 0.001. RBC transfusions containing 29.7 +/- 18.3 mmol L(-1) of potassium (RBC stored for up to 28 days) did not cause clinical or biochemical changes and reduced donor exposures by 70.2%, compared to transfusions containing 19.8 +/- 12.3 mmol L(-1) of potassium (RBC stored for up to 3 days), P < 0.001. In conclusion, RBC stored for up to 28 days safely reduced donor exposures in premature infants. PMID- 16359419 TI - Diagnosis of transfusion-associated graft-vs.-host disease: the importance of short tandem repeat analysis. AB - Transfusion-associated graft-vs.-host disease (TA-GvHD) can occur following transfusion of blood products containing immunocompetent lymphocytes, usually from HLA homozygous donors, into immunocompromised patients sharing one HLA haplotype with the donor. The diagnosis of TA-GvHD may be delayed due to the initial nonspecific clinical features involved. Investigations to detect the presence of donor-derived cells in the blood and/or affected tissues of the recipient are essential to confirm the diagnosis. We report the investigation of suspected TA-GvHD using short tandem repeat (STR) analysis, to detect the presence of donor cells (chimerism), in an immunocompetent patient admitted for coronary artery bypass surgery. Peripheral blood and skin biopsies (from affected and nonaffected sites) from the patient and peripheral blood samples from the implicated donors were taken for HLA typing and STR analysis. STR analysis revealed the presence of donor material in the patient's peripheral blood sample and in DNA extracted from the affected skin biopsy but not the unaffected biopsy, suggesting lymphocytes from this donor were responsible for the development of TA GvHD. Furthermore, HLA typing results supported the diagnosis of TA-GvHD. These data demonstrate the use of STR and HLA analysis as effective tools in the diagnosis of TA-GvHD. PMID- 16359420 TI - Donors' blood group declaration before donation can be used as a tool for electronic crossmatching. AB - Electronic crossmatching (E-XM) is used to detect ABO incompatibility. In developing countries, as many of the donations are from first-time donors, it is difficult to guarantee the accuracy of the ABO/Rh label on these units to use them for E-XM. This problem was overcome with a new software 'hemosoft', using donors' blood group declaration before donation as a tool for E-XM. During registration, donors either declare their blood group or give no comment. For, ABO/Rh grouping, either two results from different donations or only one in concordant with the declaration before donation is needed. If there is a conflict, second typing is performed from the unit segment. If donors give no declaration, two different technicians perform typing, one from the sample tube and the other from the unit segment. Of 18,618 donations performed, 640 (3%) were repeated and the rest were first-time donations. In 16,327, typing was performed once, as the blood group declaration and the typing results were identical. In 2407, grouping was performed twice, as donors gave no declaration or conflicts between declaration and typing results were found. No labelling or wrong unit release errors were detected in units donated, typed and labelled in our centre. In 26,402 donations, 16,314 (61.8%) E-XMs were performed. No major haemolytic transfusion reaction was recorded. Donors' ABO/Rh declaration before donation can be used as a tool for E-XM, instead of the requirement for serological confirmation or a second donation to guarantee grouping. PMID- 16359421 TI - Novel method for screening for the presence of haemoglobin S in blood. AB - A high-performance liquid chromatography (HPLC) method designed for the measurement of haemoglobin (Hb)A(1c) in blood was investigated for use as a screening method for the presence of HbS in blood. In the Bio-Rad VARIANT II HbA(1c) method, HbS was found to have a specific retention time and percentage Hb value that enabled the detection of HbS in blood. Other Hb variants did not have the same combination of retention time and percentage Hb as HbS. The HPLC method was superior to the HbS solubility test in ease of performance and readability. Also, the proposed method showed less interference than the solubility test and could be used with samples from all age groups. The proposed method takes 3 min per sample to perform and is thus suitable for large-scale screening. PMID- 16359422 TI - Genotyping of the Kidd blood group with allele-specific oligodeoxynucleotides coupled to fluorescent microspheres. PMID- 16359423 TI - Parental support, psychological control, and behavioral control: assessing relevance across time, culture, and method. PMID- 16359424 TI - Parsing parenting: refining models of parental influence during adolescence. PMID- 16359425 TI - A conversation about data presentation. PMID- 16359426 TI - Determinants of outcome in childhood epilepsy. AB - Possible outcomes with regard to seizures include remission (i.e. no seizures without drug treatment), conditional remission (i.e. no seizures under treatment) and treatment resistance. Several factors have been identified which are associated with favourable or unfavourable outcomes. These include syndrome diagnosis (benign vs severe epilepsy syndromes), seizure types (kind and number), severity of epilepsy, response to treatment (immediate vs delayed, monotherapy vs combinations), and concomitant neurological and psychiatric disorders. However, the quality of disease management by caretakers also has an important influence on the outcome. To determine full remission in patients who are seizure-free with treatment, antiepileptic drugs need to be tapered; generally speaking, this seems to be less risky in children than in adults. However, even if remission without treatment has been reached, an elevated risk of seizures may persist compared with that of the general population. The outcome of epilepsy should not be considered with respect to seizures alone, but should also include more global aspects of performance and quality of life. These may depend on the causes of epilepsy as much as, or more than, on the seizure disorder itself, but may also be related to treatment. PMID- 16359427 TI - Issues in paediatric epilepsy. PMID- 16359428 TI - Absences in adult seizure disorders. AB - From a phenomenological point of view, absence seizures refer to any type of epileptic event characterized by loss of awareness and responsiveness. However, a phenomenological definition of absence seizures is hampered by difficulties in conceptualizing consciousness and needs to take into account the electroclinical features and anatomical basis of seizures. Sensu stricto, absence seizures are defined as an electroclinical pattern of generalized 3/s spike-wave activity accompanied by loss of awareness and responsiveness. Although there is general belief that absences are most often encountered in childhood or early adolescence, a considerable number of patients suffer from absences late into adulthood, which are often refractory to antiepileptic drugs. These patients generally fall into one of two groups. The first group consists of those who have more or less typical absences in childhood or adolescence which continue into adulthood, and may be accompanied by other generalized seizures or neurological impairment. Few data are available addressing whether the seizure semiology of the absences in these patients changes over time or not. The second class of patients corresponds to those in whom absences emerge for the first time in adulthood or even in the senium. These need to be carefully distinguished from later aggravation or semiological transformation of undiagnosed childhood epilepsies and from certain complex focal seizures originating from the frontal or temporal lobe. A wide range of aetiologically and anatomically diverse processes may lead to the appearance of epileptic absences. It is important to note that absences also vary in their intensity, their impact on the ability of the subject to respond and react, and in particular in their natural history and their response to antiepileptic medication. PMID- 16359429 TI - Behaviour, cognition and epilepsy. AB - Cognitive and behavioural impairments have been observed as a consequence even of single seizures. In individuals with high seizure frequency, such impairments may accumulate and have a much greater impact on daily life than hitherto suspected. In addition, the risk of behavioural impairments is increased for some seizure types, such as secondary generalized seizures. Moreover, for all epilepsy types, increased risk is associated with persistent or poorly controlled seizures. Clinical studies show that cognitive impairments induced by seizures are reversible for most seizure types when seizures are controlled adequately. Additionally, for some seizure types there may be a kind of time window within which impairments are reversible. Exceeding the time window may result in irreversible impairment. These studies suggest that antiepileptic drug treatment can thus protect against such secondary behavioural impairments or at least correct these when seizures are controlled. This emphasizes the need to achieve complete and early seizure control. On the contrary, all antiepileptic drugs have a detrimental effect on the central nervous system and may affect cognitive function, behaviour and mood to some extent. Some treatments may undo the beneficial effects of antiepileptic drug treatment by inducing new or other cognitive impairments. This once more illustrates the need for the emphasis of clinical practice to evolve from mere seizure control to a more comprehensive approach, in which the prevention of central cognitive effects and effects on daily life of both seizures and drugs is given due attention. Optimal management requires a careful balance between, on the one hand, the desire to reach early and maximal seizure control and, on the other, the need to avoid tolerability problems related to cognitive and behavioural impairments. This article reviews how this balance can be achieved for older and newer antiepileptic drugs. PMID- 16359430 TI - Progress in pharmaceutical development presentation with improved pharmacokinetics: a new formulation for valproate. AB - Successful long-term treatment of patients with epilepsy requires selection of an appropriate antiepileptic regimen, optimal dosing and patient compliance. Recent advances in our understanding of the biological basis of epilepsy and in the choice of treatment options are transforming the global management of these patients. If the achievement of seizure freedom remains the primary goal of any antiepileptic treatment, issues associated with drug acceptability and tolerability, and with quality of life of patients, have gained increasing attention as major determinants of ultimate therapeutic success. Sustained release formulations of antiepileptic drugs can be very helpful in achieving treatment objectives. Stable serum levels without marked peak-to-trough fluctuations, reduced frequency of dosing and the possibility of dosing flexibility may all improve compliance, patient satisfaction and ultimately quality of life. The efficacy of sodium valproate for the treatment of most types of epilepsy has been demonstrated extensively and this drug remains the mainstay of treatment for many clinical situations. Among the various valproate formulations, extended-release tablets have shown improved patient compliance and satisfaction. However, the tablet size and the limited dosing flexibility could be unsuitable for individualized treatment in special populations such as children, the elderly and patients with swallowing difficulties. A new sustained release formulation of sodium valproate consisting of tasteless microspheres that can be sprinkled on semi-solid food such as yoghurt or jam has been developed. A stick pack presentation allows individualized dosing and greater convenience. PMID- 16359432 TI - Molecular modelling of the androgen receptor axis: rational basis for androgen receptor intervention in androgen-independent prostate cancer. AB - Androgen depletion in combination with antiandrogenic agents is initially highly effective for treating prostate cancer, and is the recommended treatment for more advanced or higher-grade tumours. However, many tumours eventually become insensitive to androgens, even though the androgen receptor (AR) continues to be expressed. Computational chemistry combined with structural analysis of nuclear receptors and determination of binding affinities of natural and designed coregulators (coactivators and corepressors) provides the theoretical framework for the rational design of novel therapeutic agents directed at the AR. Adding alternative groups to various sites throughout the receptor can alter the conformation of the molecule and its functional binding with coactivators or corepressors. Possible molecules can be identified thoroughly and systematically using intelligent high-throughput screening and FASTrack chemistry (three dimensional crystallography). Applying these techniques should eventually result in therapeutic agents for androgen-independent prostate cancer that can block binding of AR coactivators while simultaneously increasing binding of AR corepressors. PMID- 16359433 TI - Functional genomics and transcriptomics of prostate cancer: promises and limitations. PMID- 16359434 TI - Targeted discovery tools: proteomics and chromatin immunoprecipitation-on-chip. AB - Despite the availability of several completely sequenced genomes, we are still, for the most part, ignorant about how genes interact and regulate each other within a given cell type to specify identity, function and cellular memory. A realistic model of cellular regulation based on current knowledge indicates that many interacting networks operate at the epigenetic, transcriptional, translational and post-translational levels, with feedback between the various levels. Protein-protein and protein-DNA interactions help to define which genes may be activated in a particular cell, and determine whether external cues cause activation or repression. New technologies, e.g. proteomics using mass spectrometry, high-density DNA or oligonucleotide microarrays (chips), and chromatin immunoprecipitation (ChIP), provide new and exciting tools for deciphering the pathways and proteins controlling gene expression. Analysis of these pathways offers new insight that aids targeted drug development. PMID- 16359435 TI - Validation of molecular targets in prostate cancer. AB - As prostate cancer is not a single disease, it is important to identify the pivotal pathway in the patient being treated. The molecular environment is the site of current oncological research to define new therapeutic targets for hormone-refractory disease, offering the potential to eventually individualize treatment through stratification of pathways. Targets may be validated either phenotypically (e.g. androgen receptor, cadherin) or functionally (e.g. prostate cancer-specific genes). In addition, several other candidates are potentially suitable, while others await discovery. Important initial steps have been made in the search for prostate cancer stem cells; identifying stem cells and the stromal, hormonal, and other signalling molecules that influence their behaviour would have important implications for managing prostate cancer. Although individual therapeutic pathways might be ineffective in a particular molecular environment, combinations of approaches might be capable of producing synergistic effects. A multimodal approach thus might be the best solution. Determining where best to search for a molecular target, and validating whether the target is associated with a sufficiently aggressive malignant process to justify further study is difficult, but the potential benefits are enormous. PMID- 16359436 TI - Targeting apoptosis in prostate cancer: focus on caspases and inhibitors of apoptosis proteins. AB - Androgens are critical to the growth and differentiation of prostate epithelial cells. Removal of androgen normally results in apoptosis, but androgen independent tumours have developed mechanisms that allow cells to survive the loss of androgen. The caspases are central mediators of cell death. An important area for research involves manipulating caspases by novel mechanisms to induce apoptosis. However, such mechanisms as diethylmaleate priming are limited by an inability to selectively target tumour cells. Inhibitors of apoptosis proteins (IAPs) are recently identified anti-apoptotic caspase regulators. Each IAP homologue has a different mechanism of action. Because more than one member of the IAP family may be overexpressed in prostate cancer, successful treatment strategies will be defined by the ability to block all of the IAP expressed. Anti sense oligonucleotide strategies have been shown to decrease IAP expression and increase prostate cancer cell susceptibility to apoptotic induction, although not by mitochondrial-mediated pathways. Fully understanding the basic apoptotic pathway and its regulation in prostate cancer will lead to more targets for manipulation, which can be translated into novel therapies. This article focuses on the role of the caspases and IAP in developing a rational approach to using apoptosis as a therapeutic target. PMID- 16359437 TI - New strategies for the medical treatment of prostate cancer. AB - Androgen is a major growth factor in the normal prostate and determines the overall number of prostate cells. Metastatic prostate cancer, while initially responsive to androgen ablation, eventually becomes hormone-refractory and resistant to many treatments. Unfortunately, there are very few agents in the preclinical stage with a seemingly promising future for hormone-refractory prostate cancer (HRPC) that are actually taken through the complete drug development process, including US Food and Drug Administration approval. Many novel strategies under investigation for treating HRPC target metastatic prostate cancer cells that are neither androgen-dependent nor in the proliferative state. Examples of therapies that target this so-called "Achilles' heel" of HRPC include immune therapy, gene therapy, angiogenesis inhibition, and activation of programmed cell death. Unique properties of HRPC allow for the development of novel treatments that target prostate-specific antigen (PSA), human glandular kallikrein-2, or prostate-specific membrane antigen. An inactive prodrug with a thapsigargin analogue, a sesquiterpene lactone from the plant Thapsia garganica, is currently under investigation specifically for the targeted therapy of HRPC. Preclinical data suggest the PSA-targeting abilities of this novel therapy are associated with a nearly complete cessation of tumour growth with minimal toxicity. PMID- 16359438 TI - Therapeutic options in androgen-independent prostate cancer: building on docetaxel. AB - Of men with metastatic prostate cancer who undergo androgen ablation, 70-80% respond rapidly to therapy, as manifested by a reduction in prostate cancer related symptoms and declines in serum prostate-specific antigen (PSA) level. Unfortunately, after a median of 18-24 months, nearly all patients with metastatic prostate cancer will progress to androgen independence. Until recently the standard of care for treating hormone-refractory prostate cancer (HRPCa) was the combination of mitoxantrone and prednisone, which palliated bone pain but did not extend survival. Two randomized trials with > 1700 patients showed for the first time a survival benefit for patients with HRPC treated with chemotherapy; when compared with mitoxantrone-based therapy, docetaxel based-therapy reduced the risk of death by 20-24%. Future trials in HRPC are attempting to improve the efficacy of docetaxel by incorporating new agents targeting angiogenesis, apoptosis, and signal transduction pathways; there is promising activity for these novel combinations in phase I and II studies. Concepts are also being refined about definitions of response and progressive disease, patient eligibility criteria, and the validity of surrogate markers of efficacy and survival, as shown by changes in PSA level. PMID- 16359439 TI - Design of clinical trials in advanced prostate cancer: avoiding the dead ends. AB - Despite more than 30 years of clinical trials, investigations in prostate cancer have not succeeded in making advances comparable to those in other branches of research, such as breast cancer. Indeed, prostate cancer trials have repeatedly run into a series of "dead ends", as investigators face the problems of inadequate funding for research, treatments that result in only minimal improvements in survival, and lack of treatment options that have sufficient prospects for success. This article briefly reviews the strategies behind clinical investigations into prostate cancer over the last three decades, evaluates the pitfalls that have hindered research, and makes suggestions for the appropriate design of clinical trials that are safe and beneficial to patients while maintaining cost-effectiveness and accountability to patients and society. PMID- 16359440 TI - Middle management and nursing. PMID- 16359441 TI - Reluctant but resourceful middle managers: the case of nurses in the NHS. AB - This study counters the widely held view that middle managers have little to contribute to strategic change in health care organizations. In particular, it argues that middle managers with a nursing background that manage clinical activity should be involved in strategic change beyond mere implementation of decisions made by executive management. Constraints upon this are noted - the power of doctors and central government intervention - that means middle managers enact a semiautonomous strategic role. Antecedents for the semiautonomous role are investment in organization and management development, developing lateral organizational structures that allow middle managers to make a contribution to the development, as well as the implementation of strategy and allowing middle managers to interact with other stakeholders outside the confines of the organization. PMID- 16359443 TI - Understanding organizational culture: the key to successful middle manager strategic involvement in health care delivery? AB - AIM: The aim of this study was to explore the extent to which organizational culture influenced strategic involvement. BACKGROUND: It is crucial for the strategic management of health care that the cultural dimensions affecting health service organizations are identified and understood. How culture influences middle manager strategic involvement has not been established and therefore an understanding of the power of organizational culture is important to middle managers working in not-for-profit health care organizations. CONCLUSION: It was demonstrated that strong organizational culture predicted strategic involvement, and supported the importance of middle managers remaining strategically involved in the development of new organizational strategic initiatives. Findings have implications for strategic management in health service organizations. PMID- 16359442 TI - The effect of structural empowerment and perceived organizational support on middle level nurse managers' role satisfaction. AB - BACKGROUND: The restructuring of Canadian health care organizations during the past decade has reduced the visibility of nursing leadership. This has resulted in job conditions that have disempowered nurse managers and influenced their ability to create positive work environments, mentor potential nurse leaders, and gain satisfaction in the leadership role. These conditions threaten the retention of a cadre of high quality nurse leaders in today's chaotic health care setting. OBJECTIVE: The purpose of this study was to examine the relationship between structural empowerment and perceived organizational support and the effect of these factors on the role satisfaction of middle level nurse managers. METHOD: A secondary analysis was conducted as part of a larger study of 126 middle level nurse managers working in Canadian acute care hospitals, randomly selected from the Ontario provincial registry. Eighty-four nurse managers responded to a questionnaire mailed to their home addresses. RESULTS: Structural empowerment was positively associated with middle level nurse managers' perceived organizational support. The combination of empowerment and perceptions of organizational support were significant predictors of middle level nurse managers' role satisfaction. CONCLUSIONS: The findings support R.M. Kanter's (1977, 1993; Men and Women of the Corporation. Basic Books, New York) contention that empowering work conditions have an impact on employees' feelings of support and sense of accomplishment at work. Positive perceptions of organizational support may play an important role in retaining current middle managers, and possibly attracting future leaders to management positions. PMID- 16359444 TI - Social identity and self-enactment strategies: adapting to change in professional manager relationships in the NHS. AB - Significant variation has been identified concerning the impact of management agendas on health professionals in the National Health Service. While attempts to explain these responses among health professions have identified a range of factors to account for such differences, existing research is still unable to explain the sources of variation that can occur within professional groups. Current approaches to studying professional-management relations either attempt to explain variation between organizational contexts and subgroups at a collective level of analysis or they struggle because of the absence of a theoretical framework capable of integrating social, psychological and contextual elements that can explain the sources and purposes of differentiated individual and group behaviours. In the present study, we argue that if we are to go beyond the mere mapping of broad outcomes, we need to develop an understanding of how the effects of imposed managerial agendas on health professional groups come to evolve and take shape. In particular, this requires us to identify a social psychological process framework that can help explain how initiatives aimed at managerializing roles and responsibilities affect the professional employees' understandings of group membership and their relations with hospital management. By using interview data from a group of 18 hospital doctors and through a comparative grounded analysis we show how a social identity perspective may provide a suitable framework to develop verifiable explanations of this process and its implications for nurse managers. PMID- 16359445 TI - The role of the middle manager in the implementation of evidence-based health care. AB - The present study reflects on the role of the middle manager in the implementation of what has become known as evidence-based health care. This movement advocates that clinical practice is continually informed by the results of robust research and evidence. In our work exploring the complexity of ensuring that practice is informed by evidence we have found that general managers have relatively little influence when compared with clinicians especially doctors. We argue that local professional groups work together in communities of practice, which are frequently uniprofessional. These boundaries affect the motivations for seeking improvement and upgrading and the way evidence and knowledge is perceived and interpreted. We argue that if the quality of health care is to be improved, we need to understand the complex historically and contextually informed interactions between different professional groups and to design diffusion strategies that acknowledge this complexity. PMID- 16359446 TI - Nurse bullying: organizational considerations in the maintenance and perpetration of health care bullying cultures. AB - AIM: To examine bullying within nursing from a micro-sociological perspective and elucidate interactive mechanisms contributing to its causes and continuation within the nursing profession. BACKGROUND: The paper is part of a doctoral research study into bullying within nursing. It considers issues pertinent to management, and in the role of negotiated interactions within the National Health Service when dealing with bullying problems. The complex dynamics involved can be problematic for management when dealing with bullying, while often managers have been targets of bullying themselves and not infrequently accused of it. Features of bullying activity are explored, along with issues of target and bully awareness, a central feature in bullying negotiations. Issues of awareness and emergence of bullying behaviour have been identified through vignettes and unstructured interviews, and the research has identified complex interactive events in the creation and maintenance of nurse bullying activity. It is hoped that with a clearer understanding of such mechanisms and manifestations that bullying in the workplace can be reduced or eliminated. The paper is of practical use to nurse managers in illuminating such mechanisms and bringing bullying awareness to the fore. Such activity is ultimately damaging to the organization in both cost and time; and significant for professional practice by its impact upon the nurse and their work in supportive and safe environments. It will also to allow managers to consider their own practice and reactions to bullying activity within the profession. CONCLUSIONS: The overall findings from the research point strongly to bullying activity being essentially 'learned behaviour' within the workplace rather than any predominantly psychological deficit within individual perpetrators and targets. PMID- 16359447 TI - Workplace health promotion: the role and responsibility of health care managers. AB - The World Health Organization's (WHO) Ottawa Charter for Health Promotion in 1986 provided the catalyst from which the Health Promoting Workplace movement emerged. Here, an extensive review of the available workplace-related health literature provides the basis for critical discussion and recommendations for health care managers. The findings suggest that health care managers, who practice in all health service settings, should be aiming to initiate and promote radical health promotion reform as set out in the WHO settings-based movement. Developing and implementing sustainable health promotion-orientated and organization-wide healthy workplace policy initiatives represent the most effective way for health care managers to directly benefit from the desirable outcomes that come from creating and maintaining a healthy workforce. PMID- 16359448 TI - Effects of introducing a critical path method to standardize treatment and nursing for early discharge from acute psychiatry unit. AB - This research aimed to examine whether early discharge can be performed by nurses of a ward participating in the process of critical path method supporting with offer and evaluation of standard and efficient medical treatments and nursing cares. In order to enrich the system that performs hospitable medical treatment and nursing cares standardized at the psychiatry acute term and to prevent chronic, critical path method was introduced into the acute term ward, then the investigation for nursing job satisfaction that was important to establish critical path method was conducted. By introducing critical path method, it became possible to assess the patients in standard, and the subject of business or a system became clear towards early discharge. It became possible to tie up this subject to a future improvement. Consequently, as for comparison of the patient who applied the schedule table of critical path method at the first and second term, the latter decreased the average length of hospitalization period significantly. The satisfaction of the task requirement, which was important to maintain systems, was decreased by the introduction of critical path method but the score of autonomy in satisfaction increased significantly by the improvement of usage of critical path method. From these, it was suggested that the introduction of critical path method in the acute-care unit psychiatry had possibility to aim at standardization of treatment and nursing and patients' early discharge from hospitals. PMID- 16359449 TI - Nursing and history. PMID- 16359450 TI - Gender and professional identity in psychiatric nursing practice in Alberta, Canada, 1930-75. AB - This paper examines gender-specific transformations of nursing practice in institutional mental health-care in Alberta, Canada, based on archival records on two psychiatric hospitals, Alberta Hospital Ponoka and Alberta Hospital Edmonton, and on oral histories with psychiatric mental health nurses in Alberta. The paper explores class and gender as interrelated influences shaping the work and professional identity of psychiatric mental health nurses from the 1930s until the mid-1970s. Training schools for nurses in psychiatric hospitals emerged in Alberta in the 1930s under the influence of the mental hygiene movement, evolving quite differently for female nurses compared to untrained aides and male attendants. The latter group resisted their exclusion from the title 'nurse' and successfully helped to organize a separate association of psychiatric nurses in the 1950s. Post-World War II, reconstruction of health-care and a de institutionalization policy further transformed nurses' practice in the institutions. Using social history methods of analysis, the paper demonstrates how nurses responded to their circumstances in complex ways, actively participating in the reconstruction of their practice and finding new ways of professional organization that fit the local context. After the Second World War more sophisticated therapeutic roles emerged and nurses engaged in new rehabilitative practices and group therapies, reconstructing their professional identities and transgressing gender boundaries. Nurses' own stories help us to understand the striving toward psychiatric nursing professionalism in the broader context of changing gender identities and work relationships, as well as shifting perspectives on psychiatric care. PMID- 16359451 TI - Welfare state and women's work: the professional projects of nurses and occupational therapists in Sweden. AB - In this article we explore how Swedish welfare politics within health-care and rehabilitation has opened up a space for nurses' and occupational therapists' professional projects. Using historical data, an analysis of the policy-making process behind welfare programs central to the professionalization of nursing and occupational therapy is presented. The time period covered is, in the case of nurses, the larger part of the twentieth century, while the modern history of occupational therapists first began in the 1940s. Special emphasis is placed on the corporative nature of the Swedish welfare state and the professional strategies utilized by nurses and occupational therapists in their struggle for jurisdiction. In the article, politicization is identified as a core strategy by which female-dominated welfare state occupations in Sweden have tried to gain influence over the welfare policy-making process and their occupations' jurisdiction. PMID- 16359452 TI - Prelude to specialization: US cancer nursing, 1920-50. AB - This study used historical research methodology to assess the work of US nurses caring for patients with cancer from 1920 to 1950. Primary sources, obtained from several US archives, included nursing procedure books, student nurses' lecture notes, hospital and nursing annual reports, and meeting minutes. Secondary sources included journal articles and textbooks. The aim was to document the clinical work of nurses caring for patients with cancer and assess this work for evidence of emerging specialization in cancer nursing. Following a review of cancer in the 1920s, nursing education specific to cancer, nursing care related to specific cancer therapies, and the practice of concealing a diagnosis of cancer, were examined. Nurses were consistently educated in cancer nursing, but their cancer education became more focused over the decades. Nurses were closely involved with cancer patients as they monitored their patients' radium or assisted patients following radical surgery. The issue of concealing the diagnosis of cancer was problematic to some nurses. This paper reveals the nature of nurses' hospital work with cancer patients and demonstrates a core body of cancer nursing knowledge. The emergence of cancer nursing as a specialty is presented. PMID- 16359453 TI - Nursing and care for the aged in Victoria: 1950s to 1970s. AB - In the state of Victoria, Australia in the late 1950s and early 1960s, restorative treatment was introduced into the state-subsidized benevolent homes, and they were reclassified as geriatric hospitals. In the process, the nursing care of incapacitated old people was identified in terms of particular skills and knowledge, and specific forms of training were established for nurses at two levels of training: nurses' aides and supervisory nurses with a post-basic qualification. These institutional changes were complemented by the introduction of a nursing role into the body responsible for overseeing care for the aged in Victoria. Little was made of this opportunity to establish a leading role for nurses in the field of care for the aged. However, this episode raises fertile questions for further research into the development of nursing in Australia, as a set of practices divided along the lines of acute and chronic illness. PMID- 16359454 TI - The 'Manchester scheme': a study of the Diploma in Community Nursing, the first pre-registration nursing programme in a British university. AB - The Diploma in Community Nursing at the University of Manchester began in 1959, after a long period of planning and negotiation. It was the earliest pre registration nursing education programme in a British university. The paper reports on a historical study which examined its foundation and development. The history of the 'Manchester Scheme', as it came to be known, is placed into the context of wider reforms and modifications in British nursing education. The methods used for the study were a broad-based literature review, examination of material from the 1950s to the present day, five oral history interviews with some of those who were influential in developing the diploma, and the interpretation of relevant documentary source materials. The Diploma in Community Nursing at the University of Manchester was important and innovative in its time, combining as it did nurse and health visitor training with a university-based education. The paper traces the struggle to establish the course, which was largely due to the efforts of one charismatic individual, Fraser Brockington, Professor of Social and Preventive Medicine. It then goes on to examine the reforms and modifications the programme underwent from 1959 to 1969, as it was developed into the prototype for the 'Bachelor of Nursing Degree'. PMID- 16359455 TI - Influences on the early training of nurses in Denmark. AB - This paper attempts to uncover some of the factors that may have influenced the choice of model for the systematic training of nurses in Denmark. From a historical analysis of selected historical literature, nursing magazines, and archival documents, three themes emerge: the interest from (and of) the medical profession, the strategy on nurse training of the Danish Nurses' Organization, and the societal context of the time. Despite there being a Deaconess Institution in Copenhagen from 1863, it was ultimately the Nightingale model that formed the basis for the training programme. Findings suggest that the Nightingale influence within the medical establishment and among leaders in Danish nursing played a vital role in this choice. As well, the Danish Nurses' Organization actively promoted their preferred 3-year training model, which had strong similarities to the Nightingale model. Changes in family and social life after the 1864 defeat in the Prussian war fostered a growing demand for the liberalization of women's rights and a need for increased professionalization in the nursing and caring function. PMID- 16359456 TI - 'Interminable hell': Hiroshima's nurses remember the atomic bomb. PMID- 16359457 TI - Virtual reality or real virtuality: the space of flows and nursing practice. AB - The use of virtual environments for the provision of health-care is on the increase, and with each new development brings debates about their impact on care, nursing and nursing practice. Such environments offer opportunities for extending care and improvements in communication. Others believe these developments threaten aspects of nursing they hold sacrosanct. This paper explores the development of an assemblage of computer networks, databases, information systems, software programs and management systems that together work to manage health-care in Australia, namely casemix. We contend that spatial theories on network society show how this assemblage co-ordinates and operates to manage care. We discuss how this assemblage affects care and suggest that changes in organisation may be a part of the shift in how bodily organisation occurs more generally, but more specifically in health-care. We also suggest how nurses are enrolled in and by such networks, leading to transformation in nurses' practices. Finally, we argue that using spatial forms of analysis allows an interpretation of such assemblages that may account for their strengths and their shortcomings. PMID- 16359458 TI - Geography and nursing: convergence in cyberspace? AB - During the last 3 years the interface between geography and nursing has provided fertile ground for research. Not only has a conceptual emphasis on space and place provided nurse researchers with a robust and subtly different way to deconstruct and articulate nursing environments, but also their studies have provided a much needed focus on certain areas of health-care, and in particular clinical practice, not currently prioritized by health geographers. We argue that, as something that is forcing fundamental re-considerations of the nature of both nursing and geography, cyberspace is a particularly important phenomenon that lies comparatively under-researched at this interface. To encourage some interest in researching nursing and cyberspace through a geographical lens, and at least to showcase a range of potentially useful and transportable concepts, we provide an overview of some of the key debates pertaining to cyberspace developed by human geographers, and make some initial and tentative connections to nursing. PMID- 16359463 TI - A case of "migralepsy". AB - Migraine and epilepsy are common neurological conditions that may share a pathophysiologic and genetic basis. The following case is presented to illustrate key aspects of their relationship. PMID- 16359465 TI - A case of linear sebaceous nevus syndrome. PMID- 16359464 TI - Analysis of genetically complex epilepsies. AB - During the last decade, great progress has been made in the discovery of genes that influence risk for epilepsy. However, these gene discoveries have been in epilepsies with Mendelian modes of inheritance, which comprise only a tiny fraction of all epilepsy. Most people with epilepsy have no affected relatives, suggesting that the great majority of all epilepsies are genetically complex: multiple genes contribute to their etiology, none of which has a major effect on disease risk. Gene discovery in the genetically complex epilepsies is a formidable task. It is unclear which epilepsy phenotypes are most advantageous to study, and chromosomal localization and mutation detection are much more difficult than in Mendelian epilepsies. Association studies are very promising for the identification of complex epilepsy genes, but we are still in the earliest stages of their application in the epilepsies. Future studies should employ very large sample sizes to ensure adequate statistical power, clinical phenotyping methods of the highest quality, designs and analytic techniques that control for population stratification, and state-of-the-art molecular methods. Collaborative studies are essential to achieve these goals. PMID- 16359466 TI - Neurocutaneous syndromes and epilepsy-issues in diagnosis and management. AB - Epilepsy may be seen as a feature of many of the neurocutaneous syndromes. The challenge lies within the diagnosis of the specific disorder and ultimately control of the epilepsy. Tuberous sclerosis is the most common of the disorders with a frequency of 4.9/100,000. An autosomal-dominant condition, diagnostic features may be unclear under 2 years of age. Population studies suggest a prevalence of epilepsy of 78%, the majority presenting under the age of 12 months, with a high association between the occurrence of seizures and the presence of learning disability. Although an apparent multifocal disease, surgery may have a role to play where seizures are demonstrated to probably arise from a single tuber. Other less common neurocutaneous syndromes also have a high prevalence of epilepsy in association with cerebral malformations; unilateral or lobar malformations should be referred early for surgical consideration. Neurofibromatosis is the second most common of the disorders but the prevalence of epilepsy in this population is relatively low; in addition, a greater proportion may be easier to treat with medication. PMID- 16359467 TI - Genetic epidemiology of epilepsy or what do we tell families? AB - Over the past few decades, epidemiologic and molecular research has transformed the field of epilepsy genetics. In this review, we discuss the ways in which accumulating evidence on the genetics of epilepsy and febrile seizures can inform health care practitioners advising patients and families with epilepsy. We will review the epidemiologic data from twin and family studies, and illustrate how it can be used to guide genetic counseling. Although there have been many exciting advances in the last few decades-both molecular and epidemiologic-what we have learned has not appreciably changed what we tell families, and what we tell them can remain reassuring. PMID- 16359468 TI - Response to antiepileptic drug therapy: winners and losers. AB - Recent evidence suggests that the vast majority of people with epilepsy will respond to their first or second treatment regimen or will be refractory de novo. Two patients with symptomatic epilepsy secondary to underlying cortical dysplasias are presented to illustrate these different scenarios. Clinical observations can provide insights to the processes underlying pharmacoresistance. PMID- 16359469 TI - Genetics of drug resistance. AB - Drug resistance in epilepsy affects about a third of patients and is an important clinical problem, associated with increased morbidity and mortality. It is important to consider carefully the definition of drug resistance. Recent interest in the field has focused on the potential molecular mechanisms underlying drug resistance. Environmental and seizure-related acquired causes are likely to contribute to the multifaceted basis of resistance in most cases. Genetic causes have attracted particular attention, partly because they may allow prediction of drug resistance and, potentially, rational treatment strategies. Gene mutations, however, are unlikely to cause many cases of drug resistance. However, common variation in genes probably will turn out to generate an important contribution to drug resistance phenomena. Associations between common variations in a number of genes and clinical drug resistance have now been published. However, to date, none of these associations has been unequivocally replicated by others to the extent that the original association has been accepted. Some of these associations are considered. Despite this apparently uninspiring record, the genetics of drug resistance are likely to prove productive in the near future, but their pursuit will require painstaking studies and multicenter collaboration. PMID- 16359470 TI - A case of extended spectrum GEFS+. AB - A 10-year-old boy developed febrile convulsions at age 2 years, and multiple types of nonfebrile generalized seizures over the ensuing months and years. Gestation, birth, and early development were normal, as were initial EEG and brain imaging studies. By age 5 years, he had developed behavioral difficulties, and the EEG demonstrated both interictal and ictal generalized epileptiform patterns, as well as mild background slowing. Seizures were poorly controlled with multiple medication trials. There was a strong family history of clinically benign febrile seizures in the maternal lineage. Genetic analysis revealed a novel mutation in the voltage-gated neuronal sodium channel SCN1A. This patient's relatively malignant clinical course is consistent with "extended spectrum" generalized epilepsy with febrile seizures plus (GEFS+), and along with the family history illustrates the phenotypic variability of this disease. PMID- 16359471 TI - Neonatal epilepsy syndromes and generalized epilepsy with febrile seizures plus (GEFS+). PMID- 16359472 TI - Refractory calculation-induced idiopathic generalized epilepsy: a case report and review of the literature. AB - We report a case with calculation-induced idiopathic generalized epilepsy (IGE) that, unlike most patients with IGE, was refractory to medications. This patient had a family history of (1) a similar condition in a relative of hers who, however, did not have identical manifestations, and (2) a mother who had migraine. Our observations illustrate that the occurrence of IGE in families usually follows rather complex patterns of inheritance and that some of them can be refractory to therapy. PMID- 16359473 TI - Neonatal epilepsy syndromes and GEFS+: mechanistic considerations. AB - Genetic analyses of familial epilepsies over the past decade have identified mutations in several different ion channel genes that result in neonatal or early onset seizure disorders, including benign familial neonatal convulsions (BFNC), generalized epilepsy with febrile seizures plus (GEFS+), and severe myoclonic epilepsy of infancy (SMEI). These genes encode voltage-gated Na+ channel subunits (SCN1A, SCN2A, SCN1B), voltage-gated K+ channel subunits (KCNQ2, KCNQ3), and a ligand-gated neurotransmitter receptor subunit (GABRG2). While the opportunity to genotype patients for mutations in these genes can have an immediate and significant impact on our ability to diagnose and provide genetic counseling to patients, the ultimate goal is to use this molecular knowledge to develop effective treatments and cures for each disorder. This will necessitate elucidation of the molecular, cellular, and network mechanisms that translate ion channel defects into specific epilepsy phenotypes. The functional analysis of epileptogenic channel mutations in vitro and in vivo has already provided a vast amount of raw biophysical data, but attempts to interpret these data to explain clinical phenotypes so far appear to raise as many questions as they answer. Nevertheless, patterns are beginning to emerge from these early studies that will help define the full scope of the challenges ahead while simultaneously providing the foundation of future efforts to overcome them. Here, I discuss some of the potential mechanisms that have been uncovered recently linking mutant ion channel genes to neonatal epilepsy syndromes and GEFS+. PMID- 16359474 TI - Familial temporal lobe epilepsy with auditory features. PMID- 16359475 TI - Genetic focal epilepsies: state of the art and paths to the future. AB - The concept of genetic focal epilepsies is relatively new as compared to awareness of the importance of genetic factors in the generalized epilepsies. However, in the past decade, there has been increasing recognition of families with dominantly inherited partial epilepsies. Better definition of the phenotypes allows identification of distinct syndromes. The main familial focal epilepsies are autosomal-dominant nocturnal frontal lobe epilepsy (ADNFLE), familial mesial TLE (FMTLE), familial lateral TLE (FLTLE), and familial partial epilepsy with variable foci (FPEVF). The only genes identified so far are those for ADNFLE and FLTLE. In these disorders, functional studies are the next step and could provide advances leading to clarification of the pathophysiology as well as to new therapeutic strategies. At present, we can provide genetic counseling and a more accurate prognosis for most of the familial focal epilepsies. Greater awareness of the genetic basis in this group of disorders by the treating physicians is essential for identification of new families. This will allow further linkage studies, candidate gene screening, and identification of new genes, which will hopefully result in genetically based prevention and treatment. PMID- 16359477 TI - Effect of mixing methods on mechanical properties of alginate impression materials. AB - BACKGROUND: A commercial mechanical mixer is available to make the mixing of alginate more convenient and more consistent for the practitioner; however, there is very little information on the mechanical properties of alginate mixed with this device as compared with hand mixing. PURPOSE: To compare the mechanical properties of alginate impression materials mixed with a mechanical mixer (Alginator II, Cadco) and hand mixing. MATERIAL AND METHODS: Three alginate impression materials (Identic, Jeltrate, and Kromopan) were tested. Strain in compression, elastic recovery, and compressive strength were measured according to ANSI/ADA specification no. 18-1992; tear energy was measured using a pants tear test. Five specimens were prepared for each group with 12 groups for the mechanical mixer and 12 groups for hand mixing, for a total of 120 specimens. A two-way analysis of variance and Fisher's PLSD test at the 0.05 level of significance were used to analyze the data. RESULTS: There were statistically significant differences in properties among the materials, but mixing technique had no statistically significant effect on strain in compression and tear energy. CONCLUSION: The mechanical mixer improved elastic recovery and compressive strength of the alginate impression materials tested and had no effect on strain in compression and tear energy. A mechanical mixer facilitates the mixing of alginate impression materials and improves some mechanical properties. PMID- 16359478 TI - Clinical acceptability of crown margins versus marginal gaps as determined by pre doctoral students and prosthodontists. AB - PURPOSE: Marginal integrity is a very important element in evaluating a restoration; however, there is no agreement in definition of a clinically acceptable margin. The purpose of this investigation was to examine margin acceptability using an explorer versus the actual marginal gap widths at four locations on uncemented crowns on three extracted teeth using both predoctoral students and prosthodontists as evaluators. MATERIALS AND METHODS: The crown margin evaluation used 16 surfaces of four crowns fitting to three extracted caries-free teeth fitted into a dentiform. The teeth (nos. 14, 20, and 29) were prepared for a full cast (gold) crown using a chamfer finish line configuration, with some margins supragingival and others subgingival. After final impressions and working casts were made, die spacer was applied to the marginal area of the die before waxing to vary the marginal opening. The dentiform was placed in a mannequin in a supine position. Predoctoral students (N = 10) and prosthodontists (N = 9) evaluated each axial surface of each crown in the zone along the margin with an explorer and rated each surface as either "clinically acceptable" or "unacceptable." After casting, the axial marginal openings were measured with Image Pro Software using a digital microscopic image of the surface. Each participant repeated the margin evaluations 6 months later. RESULTS: Upon casting, marginal gaps ranged from 40 microm to 615 microm. The proportions of prosthodontists and of predoctoral students rating a given surface as "clinically unacceptable" were highly correlated (Spearman rank correlation = 0.81, p= 0.0001). The prosthodontists did not provide more or fewer ratings of clinical acceptability than the students, although kappa results indicated that the prosthodontists might be more consistent among themselves than the student raters. Upon re-evaluation, both groups rated between one and six of the surfaces differently than they had previously: the median number of inconsistencies was 1 for prosthodontists and 3 for predoctoral students. The prosthodontists tended to have fewer inconsistencies than the predoctoral students (0.05 < p < 0.10 Wilcoxon rank sum test), but this was not statistically significant. CONCLUSION: The data provided evidence that those surfaces associated with greater margin gaps tended to have a greater proportion of ratings of "clinically unacceptable." The proportion of prosthodontists and predoctoral students rating a margin "clinically unacceptable" were highly correlated. Prosthodontists tended to have fewer inconsistencies than predoctoral students, but that difference was not statistically significant. PMID- 16359479 TI - The effect of water absorption on acrylic surface properties. AB - PURPOSE: The aim of the present study was to determine whether an increased water content during thermal cycling of hot water-treated acrylic was associated with a reduction in surface hardness and an increased opacity or whitening of the surface. MATERIALS AND METHODS: Ten acrylic samples were treated with 30 soak cycles (cycle duration, 24 hours), using warm water (40 degrees C) and an alkaline peroxide tablet (Efferdent control group); a further ten samples were treated with boiling water (100 degrees C) and one Efferdent tablet (experimental group). Indentation hardness of the acrylic specimens was measured prior to and immediately following the completion of the warm and hot water treatments, using an automated micro-indentation system. The hydrated acrylic specimens were then allowed to air dry at room temperature (20 degrees C) and were weighed weekly until they had obtained a constant dry weight. The loss in weight of the acrylic specimens represented the maximum water absorption. RESULTS: The hot water treated specimens were much whiter than the warm water-treated specimens. The mean reduction in hardness (H(IT)) of the acrylic specimens following the treatment with hot water and alkaline peroxide tablet was 12.9%. Treatment with warm water and alkaline peroxide resulted in a slight increase in mean hardness (2.63%). There was a significant correlation between the water content of the acrylic specimens after treatment and the percentage of change in indentation hardness (r= 0.495, p= 0.026). CONCLUSIONS: The hot water treatment of the acrylic was associated with a significant reduction in hardness. We attribute the whitening and reduction in the hardness of the hot water-treated specimens to absorption of water and a disruption of the acrylic surface structure. PMID- 16359480 TI - Comparison of bond strength of a pressed ceramic fused to metal versus feldspathic porcelain fused to metal. AB - PURPOSE: To compare the debonding/crack initiation strength (D/CIS) of a low fusing pressable leucite-based glass ceramic (PC) fused to metal to a feldspathic porcelain (FP) fused to metal. MATERIALS & METHODS: As per ISO 9693:1999, 40 rectangular metal specimens (25.0 mm x 0.5 mm x 3.0 mm) were prepared. Twenty of the specimens were cast in a base metal nickel-chromium alloy (BA), and 20 were cast in a noble metal palladium-silver alloy (NA). Ten randomly selected NA and BA alloy specimens had FP applied. The remaining 10 NA and BA alloy specimens had ash-free wax patterns applied, the metal-wax complexes invested, and were pressed with a PC. The dimensions of the ceramic specimens were 8.0 mm x 1.0 mm x 3.0 mm, creating a combined metal-ceramic complex thickness of 1.5 mm. All specimens were subject to a three-point bending test at a crosshead speed of 1.5 mm/min. Fracture loads were recorded in Newtons and D/CISs calculated by the formula tau(b)=k x F(fail). RESULTS: Mean D/CISs, measured in MPa (standard deviations): NA-FP 32.56 (4.62), NA-PC 30.23 (5.06), BA-FP 30.98 (4.41), and BA-PC 31.81 (3.48). A two-way ANOVA (p > 0.05) did not demonstrate significant difference between groups. CONCLUSION: The debonding/crack initiation strength of a low fusing pressable leucite-based glass ceramic fused to metal was equivalent to that of a feldspathic porcelain fused to metal. PMID- 16359481 TI - Subjective assessment by patients of the efficiency of two denture adhesive pastes. AB - PURPOSE: This study evaluated the subjective responses of 30 denture wearers with regard to the effectiveness of two denture adhesive pastes. MATERIALS AND METHODS: Thirty patients responded to questions related to retention, chewing ability, taste, duration of adhesives in the mouth, and removal; comparing the use of two denture adhesives based on polymethylvinylether-maleic anhydride compounds or carboxymethyl cellulose. RESULTS: The denture adhesive paste based on polymethylvinylether-maleic anhydride (PVM-MA) compound was rated higher (73% and 87%) than carboxymethyl cellulose (CC) adhesive paste (60% and 37%) on chewing ability and duration in the mouth (p= 0.0001 and p= 0.0001, respectively) by experienced denture wearers in both maxilla and mandible. There were no statistically significant differences between the two denture adhesive pastes on the retention of maxillary dentures, taste, and removal of the adhesive (p= 0.08, p= 0.67, and p= 0.41, respectively). CONCLUSIONS: All subjects responded that the retention of their dentures was either a little better or much better when using either of the adhesive pastes. PMID- 16359482 TI - A survey of U.S. prosthodontists and dental schools on the current materials and methods for final impressions for complete denture prosthodontics. AB - PURPOSE: The purpose of this study was to survey members of The American College of Prosthodontists (ACP) to evaluate current materials and methods for final impressions for complete denture prosthodontics in the United States. In addition, those methods were compared with methods and materials taught in U.S. dental schools via a second survey sent to the chairpersons of prosthodontic/restorative departments. MATERIALS AND METHODS: An anonymous questionnaire was mailed to all 1762 active ACP members in the United States in 2003. A slightly modified questionnaire was also distributed to chairpersons of prosthodontic/restorative departments in the 54 U.S. dental schools. Data analysis was performed via frequency distribution and chi-square statistics. RESULTS: Nine hundred and forty-five questionnaires were returned by members of the ACP (54% return rate) and 42 questionnaires were returned by the U.S. dental schools (78% return rate). The majority of the reporting prosthodontists (88%) and dental schools (98%) use a border-molded custom tray for final impressions for complete denture prosthodontics. The most popular material for border molding was plastic modeling compound (67% of reporting ACP members, and 95% of the responding dental schools). Variability of the materials used for final impressions was observed, with the most popular materials being polyvinylsiloxane for the ACP members (36%) and polysulfide for the dental schools (64%). Statistically significant differences were found in the materials used for border molding by prosthodontists based on the time elapsed since completion of prosthodontic training. No differences were found in the materials used for impression of edentulous arches based on years of experience. Geographic location did not influence the materials and methods used by prosthodontists for complete denture final impressions. CONCLUSIONS: There was variability of the materials and techniques used for final impressions by ACP members and dental schools; however, overall there was an agreement on the materials and techniques used by prosthodontists and dental schools. Distinct trends for increasing use of polyvinylsiloxane and polyether for border molding procedures and impressions of edentulous arches were observed both in members of the ACP and in the U.S. dental schools. PMID- 16359484 TI - What prosthodontists can learn from Starbucks' 6 squares. PMID- 16359488 TI - A prosthodontic wake-up call: reaffirming the complete denture, implants alone do not solve the problems: the old principles are still important. PMID- 16359490 TI - Understanding heart development and congenital heart defects through developmental biology: a segmental approach. AB - ABSTRACT The heart is the first organ to form and function during development. In the pregastrula chick embryo, cells contributing to the heart are found in the postero-lateral epiblast. During the pregastrula stages, interaction between the posterior epiblast and hypoblast is required for the anterior lateral plate mesoderm (ALM) to form, from which the heart will later develop. This tissue interaction is replaced by an Activin-like signal in culture. During gastrulation, the ALM is committed to the heart lineage by endoderm-secreted BMP and subsequently differentiates into cardiomyocyte. The right and left precardiac mesoderms migrate toward the ventral midline to form the beating primitive heart tube. Then, the heart tube generates a right-side bend, and the d-loop and presumptive heart segments begin to appear segmentally: outflow tract (OT), right ventricle, left ventricle, atrioventricular (AV) canal, atrium and sinus venosus. T-box transcription factors are involved in the formation of the heart segments: Tbx5 identifies the left ventricle and Tbx20 the right ventricle. After the formation of the heart segments, endothelial cells in the OT and AV regions transform into mesenchyme and generate valvuloseptal endocardial cushion tissue. This phenomenon is called endocardial EMT (epithelial-mesenchymal transformation) and is regulated mainly by BMP and TGFbeta. Finally, heart septa that have developed in the OT, ventricle, AV canal and atrium come into alignment and fuse, resulting in the completion of the four-chambered heart. Altered development seen in the cardiogenetic process is involved in the pathogenesis of congenital heart defects. Therefore, understanding the molecular nature regulating the 'nodal point' during heart development is important in order to understand the etiology of congenital heart defects, as well as normal heart development. PMID- 16359491 TI - Descriptive epidemiology of anotia and microtia, Hawaii, 1986-2002. AB - The objective of this investigation was to describe the epidemiology of anotia and microtia with respect to various factors. The cases studied were all infants and fetuses with anotia or microtia identified by a population-based birth defects registry in Hawaii. The anotia and microtia rates were determined for selected factors and comparisons made among the subgroups by calculating the rate ratio (RR) and 95% confidence interval (CI). A total of 120 cases were identified, for a rate of 3.79 per 10,000 live births. The anotia and microtia rate increased during 1986-2002, although the trend was not significant (P = 0.715). Of 49 specific structural birth defects examined, four were found to be significantly more common in the presence of anotia and microtia. When compared with Caucasians, the anotia and microtia rates were higher among Far East Asians (RR 1.79, 95% CI 0.89-3.68), Pacific Islanders (RR 2.26, 95% CI 1.24-4.32), and Filipinos (RR 2.34, 95% CI 1.23-4.64). The defects were less common among females (RR 0.64, 95% CI 0.43-0.93) and more common with multiple birth (RR 3.72, 95% CI 1.66-7.33), birth weight < 2500 g (RR 3.35, 95% CI 2.04-5.30), and gestational age <38 weeks (RR 2.27, 95% CI 1.49-3.40). In conclusion, the rate for anotia and microtia increased in Hawaii during the study period. The rates for only a few structural birth defects were substantially greater than expected in association with anotia and microtia. Anotia and microtia rates varied significantly according to maternal race/ethnicity, infant sex, plurality, birth weight, and gestational age. PMID- 16359492 TI - Comprehensive screening of CREB-binding protein gene mutations among patients with Rubinstein-Taybi syndrome using denaturing high-performance liquid chromatography. AB - Mutations in the CREBBP (CREB-binding protein gene) cause Rubinstein-Taybi syndrome (RSTS). At present, however, genetic testing of CREBBP is not commonly applied in clinical settings because the currently available assays are technically and financially demanding, mainly because of the size of the gene. In the present study, we took advantage of a highly sensitive and specific, automated denaturing high-performance liquid chromatography (DHPLC) technique. First, we developed a DHPLC-based protocol to analyze the entire coding region of CREBBP. Second, we analyzed genetic samples from 21 RSTS patients using DHPLC. The coding region was amplified by 41 primer pairs, all of which have the same cycling conditions, aliquoted on a 96-well format PCR plate. In this manner, all the exons were simultaneously amplified using a single block in a PCR machine. We then wrote a computer script to analyze all the PCR amplicons generated from various portions of the CREBBP gene in a serial manner at optimized conditions determined individually for each amplicon. Heterozygous CREBBP mutations were identified in 12 of the 21 patients: five frameshift mutations, three nonsense mutations, two splice-site mutations, and two missense mutations. The resulting detection rate of 57% was comparable to the outcome of previous studies. The relatively high detection rate in the present study demonstrates the enhanced sensitivity of the DHPLC-based mutation analysis, as exemplified by mutation analyses of other genes. The implementation of similar methodologies for other dysmorphic syndromes will help medical geneticists to confirm their clinical impressions and to provide accurate genetic counseling for patients and their families. PMID- 16359493 TI - Exencephaly induction by valproic acid in the genetic polydactyly/arhinencephaly mouse, Pdn/Pdn. AB - Non-treated homozygous polydactyly/arhinencephaly (Pdn/Pdn) mouse fetuses exhibited exencephaly in 16.7% of cases. Treatment of Pdn/Pdn mice with 350 mg/kg of valproic acid (VPA) on days 8.5 and 9.5 of gestation increased the rate of exencephaly to 66.7%. The responsible gene for the Pdn mouse phenotype has been determined to be Gli3, and the suppression of Gli3 gene expression has been documented in Pdn/Pdn embryos. We investigated how the sonic hedgehog (Shh) and Fgf8 genes, the correlated genes of Gli3, are expressed in the VPA-treated exencephalic Pdn/Pdn embryos on day 10 of gestation, using whole mount in situ hybridization (WISH) and real-time PCR methods. We could not detect any alterations in Shh expression by real-time PCR, or WISH in the non-treated Pdn/Pdn and VPA-treated exencephalic Pdn/Pdn embryos. Altered Fgf8 expression patterns were observed in the commissural plate and dorsal isthmal neuroepithelium in the non-treated Pdn/Pdn embryos. We speculated that the altered expression of Fgf8 might be the result of down-regulation of Gli3 in Pdn/Pdn embryos. Fgf8 gene expression in the commissural plate and dorsal isthmal neuroepithelium exhibits wide or altered signal patterns in the VPA-treated exencephalic Pdn/Pdn embryo. From these findings, it was suggested that down regulation of Gli3 gene expression induced the altered expression of Fgf8 in the Pdn/Pdn embryos, and that VPA treatment accelerated the alterations of Fgf8 gene expression in the Pdn/Pdn embryos. It was further speculated that altered expression of Fgf8 in the commissural plate may be the fundamental cause of exencephaly, and that the synergistic effect between gene and drug shown in this experiment may explain the differences of sensitivity in the side-effects of the drug. PMID- 16359494 TI - Comparative susceptibility of newborn and young rats to six industrial chemicals. AB - To elucidate the comparative susceptibility of newborn rats to chemicals, newborn and young animals were administered six industrial chemicals by gavage from postnatal days (PND) 4 to 21, and for 28 days starting at 5-6 weeks of age respectively, under the same experimental conditions as far as possible. As two new toxicity endpoints specific to this comparative analysis, presumed no observed-adverse-effect-levels (pNOAELs) were estimated based on results of both main and dose-finding studies, and presumed unequivocally toxic levels (pUETLs) were also decided. pNOAELs for newborn and young rats were 40 and 200 for 2 chlorophenol, 100 and 100 for 4-chlorophenol, 30 and 100 for p-(alpha,alpha dimethylbenzyl) phenol, 100 and 40 for (hydroxyphenyl)methyl phenol, 60 and 12 for trityl chloride, and 100 and 300 mg/kg/day for 1,3,5-trihydroxybenezene, respectively. To determine pUETLs, dose ranges were adopted in several cases because of the limited results of experimental doses. Values for newborn and young rats were thus estimated as 200-250 and 1000 for 2-chlorophenol, 300 and 500 for 4-chlorophenol, 300 and 700-800 for p-(alpha,alpha-dimethylbenzyl) phenol, 140-160 and 1000 for (hydroxyphenyl)methyl phenol, 400-500 and 300 for trityl chloride, and 500 and 1000 mg/kg/day for 1,3,5-trihydroxybenzene, respectively. In most cases, newborn rats were 2-5 times more susceptible than young rats in terms of both the pNOAEL and the pUETL. An exception was that young rats were clearly more susceptible than their newborn counterparts for trityl chloride. PMID- 16359495 TI - Elevated susceptibility of newborn as compared with young rats to 2-tert butylphenol and 2,4-di-tert-butylphenol toxicity. AB - In order to determine the susceptibility of newborn rats to 2-tert-butylphenol (2TBP) and 2,4-di-tert-butylphenol (DTBP) toxicity, studies were conducted with oral administration from postnatal days (PND) 4 to 21 and the findings were compared with results for young rats exposed from 5 or 6 weeks of age for 28 days. In the newborn rats, specific effects on physical and sexual development and reflex ontogeny were not observed. While there were no clear differences in toxicological profiles between newborn and young rats, the no-observed-adverse effect levels (NOAELs) differed markedly. For 2TBP, clinical signs such as ataxic gait, decrease in locomotor activity and effects on liver, such as increase in organ weight, were observed and the NOAELs were concluded to be 20 and 100 mg/kg/day in newborn and young rats, respectively. Based on hepatic and renal toxicity (histopathological changes and increase in organ weight with blood biochemical changes), the respective NOAELs for DTBP were concluded to be 5 and 20 mg/kg/day. Therefore, the susceptibility of newborn rats to 2TBP and DTBP was found to be 4-5 times higher than that of young rats. PMID- 16359496 TI - Massive cystic lymphangiomas of a fetus. AB - We present a fetus with progressive massive subcutaneous lymphangiomas leading to intrauterine death. A 28-year-old woman was referred to our hospital because of a precordial cystic mass of the fetus. An ultrasound revealed lymphangiomas extending from bilateral axillae to the anterior chest wall. At 18 weeks' gestation, amniocentesis was performed and the karyotype of the fetus was found to be normal 46, XY. Thereafter the lesions increased in size gradually and spread over the body. Amniotic fluid decreased, pericardial, and pleural effusion appeared, and cardiomegaly became evident. The fetus died in utero at 25 weeks' gestation. Postmortem examination revealed a male fetus surrounded with multicystic soft masses spreading over the body, and syndactyly (left third and fourth fingers) was present. Histologically, a number of irregularly dilated lymphatics extended through subcutaneous tissues to the skeletal muscles. No communications between the cysts and the thoracic or abdominal cavity existed, and no lymphatic dilations in the viscera were confirmed. As far as we know, such conditions have rarely been reported. Considering that in previous literature, a favorable prognosis of a fetus with an atypically located (lateral cervical or non-cervical) lymphangioma with a normal karyotype has been reported, our case may be included in a distinct pathological entity. When we find a lymphangioma in a fetus, careful follow-up by ultrasound is mandatory. PMID- 16359497 TI - Spontaneous regression of congenital cystic adenomatoid malformation of the lung: longitudinal examinations by magnetic resonance imaging. AB - We report a case of large cystic adenomatoid malformation of the lung (CCAM), which occupied almost the entire left lung with a prominent mediastinal shift at 24 weeks of gestation. The volume of the lesion, determined by magnetic resonance imaging (MRI), was 27.0 cm3. Subsequent MRI and ultrasound examinations revealed a spontaneous resolution of the lesion at 32 and 36 weeks of gestation without a mediastinal shift, when the lesion volume was 12.8 cm3 and 5.6 cm3, respectively. At 37 weeks of gestation, a mature male baby weighing 2638 g with an Apgar score of 7 was delivered by elective cesarean section. A lobectomy of the left upper lobe of the lung was carried out at 3 days of age, due to an enlargement of the CCAM after birth. PMID- 16359498 TI - Stillborn baby with partial monosomy of the short arm of chromosome 5 and partial trisomy of the short arm of chromosome 20. AB - A stillborn baby with multiple malformations, including cardiac defects and cerebellar hypoplasia, is described. The abnormal features were ascribed to an unbalanced chromosome translocation, resulting in a partial deletion of the short arm of chromosome 5 and a partial trisomy of the short arm of chromosome 20. A parental balanced translocation t(5; 20)(p13.3; p11.23) was identified. The present case is the first case in Japan of monosomy of the short arm of chromosome 5 and trisomy of the short arm of chromosome 20. PMID- 16359499 TI - Is aspirin effective in diabetic patients? Yes. PMID- 16359500 TI - Is aspirin effective in diabetic patients? No. PMID- 16359501 TI - von Willebrand disease--the relevance of history. PMID- 16359502 TI - The discriminant power of bleeding history for the diagnosis of type 1 von Willebrand disease: an international, multicenter study. AB - OBJECTIVE: The aim of this study was the validation of the criteria defining a significant mucocutaneous-bleeding history in type 1 von Willebrand disease (VWD). SUBJECTS AND METHODS: To avoid selection bias, 42 obligatory carriers (OC) of type 1 VWD were identified from a panel of 42 families with type 1 VWD enrolled by 10 expert centers. OC were identified by the presence of an offspring and another first degree relative with type 1 VWD (affected subjects, AFF). A standardized questionnaire was administered to evaluate hemorrhagic symptoms at the time of first examination, using a bleeding score ranging from 0 (no symptom) to 3 (hospitalization, replacement therapy, blood transfusion). Sensitivity, specificity, diagnostic likelihood ratios, positive and negative predictive values for the diagnosis of type 1 VWD were calculated from the data collected in OC and in 215 controls. RESULTS: Having at least three hemorrhagic symptoms or a bleeding score of 3 in males and 5 in females was very specific (98.6%) for the bleeding history of type 1 VWD, although less sensitive (69.1%). None of the misclassified OC had life-threatening bleeding episodes after diagnosis. CONCLUSIONS: We suggest that the use of a standardized questionnaire and bleeding score may be useful for the identification of subjects requiring laboratory evaluation for VWD. PMID- 16359503 TI - Enhanced antiplatelet effect of clopidogrel in patients whose platelets are least inhibited by aspirin: a randomized crossover trial. AB - OBJECTIVE: We aimed to determine whether adding clopidogrel to aspirin in patients at high risk of future cardiovascular events would suppress laboratory measures of the antiplatelet effects of aspirin; and have greater platelet inhibitory effects in patients with the least inhibition of platelets by aspirin. METHODS: We performed a randomized, double-blind, placebo-controlled, crossover trial, comparing clopidogrel 75 mg day(-1) versus placebo, in 36 aspirin-treated patients with symptomatic objectively confirmed peripheral arterial disease. RESULTS: The addition of clopidogrel to aspirin did not suppress platelet aggregation induced by arachidonic acid, urinary 11 dehydro thromboxane B2 concentrations, or soluble markers of platelet activation markers (P-selectin, CD40-ligand) and inflammation (high sensitivity serum C-reactive protein, interleukin-6). Clopidogrel significantly inhibited platelet aggregation induced by ADP (reduction 26.2%; 95% CI: 21.3-31.1%, P < 0.0001) and collagen (reduction 6.2%; 95% CI: 3.2-9.3%, P = 0.0003). The greatest inhibition of collagen-induced platelet aggregation by clopidogrel was seen in patients with the least inhibition of arachidonic acid induced aggregation by aspirin [lower tertile of arachidonic acid-induced platelet aggregation: 2.8% (95% CI: -0.8 to 6.3%) reduction in mean collagen-induced aggregation by clopidogrel; middle tertile: 4.0% (95% CI: 0.4-7.6%); upper tertile 12.6% (95% CI: 4.5-20.8%); P-value for interaction 0.01]. CONCLUSIONS: The greatest platelet inhibitory effect of clopidogrel occurs in patients with the least inhibition of arachidonic acid induced platelet aggregation by aspirin. This raises the possibility that the clinical benefits of adding clopidogrel to aspirin may be greatest in patients whose platelets are least inhibited by aspirin. Confirmation in clinical outcome studies may allow these patients to be targeted with antiplatelet drugs that inhibit the ADP receptor, thereby overcoming the problem of laboratory aspirin resistance. PMID- 16359504 TI - Genetic analysis of 31 Swedish type 1 von Willebrand disease families reveals incomplete linkage to the von Willebrand factor gene and a high frequency of a certain disease haplotype. AB - BACKGROUND: The most common type of von Willebrand disease (VWD), type 1, has in only a few cases been explained by an identified causative mutation in the von Willebrand factor (VWF) gene. The ABO blood group and other modifier loci outside the VWF gene may contribute to the development of type 1 VWD. OBJECTIVES AND METHODS: Our aim was to determine whether there was genetic linkage to the VWF gene in 31 Swedish type 1 VWD families. Stringent diagnostic criteria in accordance with ISTH guidelines were used. Genetic linkage was investigated by using two highly informative dinucleotide microsatellite markers, which we have recently identified, located in introns six and 15 of the VWF gene. We also investigated the existence of common disease haplotypes and the relation between type 1 VWD and ABO blood group. RESULTS: We found genetic linkage to the VWF gene in 27 (87%) of the families. However, in four (13%) of the families, there was clearly no genetic linkage. We found the 4751A>G (Tyr1584Cys) sequence variation in exon 28, which is a common mutation in the Canadian VWD population (14.3%), in only one of the 31 families (3.2%). A possible common mutation was identified in six of the 27 (22%) families with genetic linkage. Blood group O was over represented among type 1 VWD patients. CONCLUSION: We conclude that there is linkage between the VWF gene and hereditary type 1 VWD in a majority of families. PMID- 16359505 TI - Deep-vein thrombosis rates after major orthopedic surgery in Asia. An epidemiological study based on postoperative screening with centrally adjudicated bilateral venography. AB - BACKGROUND: The incidence of postsurgical venous thromboembolism is thought to be low in Asian ethnic populations. OBJECTIVE: We studied the incidence of deep-vein thrombosis (DVT) in Asian patients undergoing major orthopedic surgery of the lower limbs. PATIENTS/METHODS: We performed a prospective epidemiological study in 19 centers across Asia (China, Indonesia, South Korea, Malaysia, Philippines, Taiwan, and Thailand) in patients undergoing elective total hip replacement (THR), total knee replacement (TKR) or hip fracture surgery (HFS) without pharmacological thromboprophylaxis. The primary endpoint was the rate of DVT of the lower limbs documented objectively with bilateral ascending venography performed 6-10 days after surgery using a standardized technique and evaluated by a central adjudication committee unaware of local interpretation. RESULTS: Overall, of 837 Asian patients screened for this survey, 407 (48.6%, aged 20-99 years) undergoing THR (n = 175), TKR (n = 136) or HFS (n = 96) were recruited in 19 centers. DVT was diagnosed in 121 of 295 evaluable patients [41.0%, (95% confidence interval (CI): 35.4-46.7)]. Proximal DVT was found in 30 patients [10.2% (7.0-14.2)]. Total DVT and proximal DVT rates were highest in TKR patients (58.1% and 17.1%, respectively), followed by HFS patients (42.0% and 7.2%, respectively), then THR patients (25.6% and 5.8%, respectively). DVT was more frequent in female patients aged at least 65 years. Pulmonary embolism was clinically suspected in 10 of 407 patients (2.5%) and objectively confirmed in two (0.5%). CONCLUSIONS: The rate of venographic thrombosis in the absence of thromboprophylaxis after major joint surgery in Asian patients is similar to that previously reported in patients in Western countries. PMID- 16359506 TI - The post-thrombotic syndrome: risk factors and impact on the course of thrombotic disease. AB - BACKGROUND: The post-thrombotic syndrome (PTS) is a frequent complication of deep vein thrombosis (DVT). Patients with recurrent ipsilateral DVT have an increased risk of PTS; other risk factors are unknown. OBJECTIVES: To establish risk factors of PTS and its impact on venous thrombotic disease. PATIENTS: We prospectively followed 406 patients after a first symptomatic DVT for a median of 60 months. Patients with recurrent DVT, a natural inhibitor deficiency, the lupus anticoagulant, cancer, long-term anticoagulation, an observation time < 18 months and DVT-recurrence prior PTS-assessment were excluded. Study outcomes were occurrence of PTS and recurrent symptomatic DVT. RESULTS: PTS was assessed after 44 +/- 23 months (mean +/- SD) using a clinical classification score. PTS developed in 176 of 406 patients (43.3%). Severe PTS was rare (1.4%). Proximal DVT was the strongest risk factor of PTS [odds ratio (OR) 2.1, 95% confidence interval (CI) 1.3-3.7]. Male gender (OR 1.6, 95% CI 1.0-2.8) and elevated D-dimer levels (OR 1.9, 95% CI 1.0-3.9) were weaker risk factors. Factor V Leiden, factor II G20210A or high factor VIII did not confer an increased risk of PTS. At 4 years, the cumulative probability of recurrence was 7.4% (95% CI 3.2-11.7) among patients with PTS when compared with 1.6% (95% CI 0-3.5; P < 0.02) among patients without PTS. The risk of recurrence was 2.6-fold (95% CI 1.2-5.9) increased when PTS was present. CONCLUSIONS: Proximal DVT, male gender, and high D-dimer levels are independently associated with the development of PTS in patients with a first DVT. Patients with PTS have an increased risk of recurrent venous thromboembolism. PMID- 16359507 TI - von Willebrand factor ristocetin cofactor (VWF:RCo) assay: implementation on an automated coagulometer (ACL). AB - BACKGROUND: VWF:RCo assay is the standard and widely used laboratory test for von Willebrand disease (VWD) diagnosis. It is hampered by high intra- and inter-assay imprecision and is time consuming. Automation may improve the assay performance and allow its routine application. OBJECTIVE: Automation of VWF:RCo on the ACL 7000 coagulometer (Instrumentation Laboratory, Milan, Italy) and its evaluation in VWD diagnosis. METHODS AND MATERIALS: Method performance determination: precision, detection limit (DL), interferences, dose-response curve. Method comparison: analysis of 105 plasma samples from normal subjects (50), VWD type 1 (24), VWD type 2 (24) and VWD type 3 (7) with ACL VWF:RCo and comparison with the reference aggregometric (AGM) method. RESULTS: ACL VWF:RCo: CVs around 10% vs. 19% of AGM method; DL: 0.08 U mL(-1); potential interferences from bilirubin, triglycerides and hemoglobin, avoided by suitable plasma dilution; high correlation with AGM VWF:RCo (Deming regression Y =-0.0277 + 1.0519X) either in normal or VWD plasmas. In VWD types 1 and 2, the VWF:RCo/VWF:Ag ratios are >0.6 or <0.6, respectively, when calculated with both AGM and ACL VWF:RCo values. CONCLUSIONS: The automated VWF:RCo on the ACL 7000 coagulometer shows precision improvement and high correlation with the reference AGM method. The test allows the diagnosis of both quantitative (VWD types 1 and 3) and qualitative (VWD type 2) forms of the disease. These results and the assay feasibility make it a suitable and reliable test for the routine diagnosis of VWD. PMID- 16359508 TI - Expression of the normal factor V allele modulates the APC resistance phenotype in heterozygous carriers of the factor V Leiden mutation. AB - BACKGROUND: Functional defects of the protein C pathway, detectable in plasma as activated protein C (APC) resistance, are a prevalent risk factor for venous thrombosis. The factor V (FV) Leiden mutation causes APC resistance by interfering with the APC-mediated inactivation of both FVa and FVIIIa. Co inheritance of FV Leiden and quantitative FV deficiency on different alleles, a rare condition known as pseudo-homozygous APC resistance, is associated with pronounced APC resistance and 50% reduced FV levels, because of non-expression of the non-Leiden FV allele. OBJECTIVES: The role of normal FV in modulating the APC resistance phenotype in carriers of FV Leiden was investigated in patients with pseudo-homozygous APC resistance and in model systems. PATIENTS/METHODS: Four functional plasma assays probing both components of APC resistance (susceptibility of FVa to APC and cofactor activity of FV in FVIIIa inactivation) were employed to compare seven clinically and genetically characterized FV Leiden pseudo-homozygotes to 30 relatives with different FV genotypes (including 12 FV Leiden heterozygotes and seven carriers of FV deficiency) and to 32 unrelated FV Leiden homozygotes. RESULTS AND CONCLUSIONS: All assays consistently indicated that FV Leiden pseudo-homozygotes are significantly more APC-resistant than heterozygotes and indistinguishable from homozygotes. Thrombin generation measurements in FV-deficient plasma reconstituted with purified normal FV and FV Leiden confirmed these observations and showed that the expression of the normal FV allele is an important modulator of APC resistance in FV Leiden heterozygotes. These findings provide an explanation for the higher thrombotic risk of FV Leiden pseudo-homozygotes when compared with heterozygotes. PMID- 16359509 TI - Proteolytic cleavage of protein S during the hemostatic response. AB - BACKGROUND: Protein S is a vitamin K-dependent protein with anticoagulant properties. It contains a so-called thrombin-sensitive region (TSR), which is susceptible to cleavage by coagulation factor Xa (FXa) and thrombin. Upon cleavage, the anticoagulant activity of protein S is abolished. OBJECTIVE: The aim of the present study was to determine whether protein S is cleaved within the TSR during activation of the coagulation system under near physiological conditions. RESULTS: In a reconstituted coagulation system containing apart from protein S only procoagulant constituents and synthetic phospholipid vesicles, protein S was cleaved at Arg60 by the FXa generated (3 mol min(-1) mol(-1) enzyme). FXa-catalyzed cleavage of protein S, however, was inhibited by factor Va and prothrombin by more than 70%. During clotting of recalcified citrated plasma in the presence of a synthetic lipid membrane, no FXa-catalyzed proteolysis of protein S was observed. Substituting platelets for phospholipid vesicles resulted both in the reconstituted system and in plasma in cleavage of the TSR. Cleavage was at Arg60 and was observed upon platelet activation, irrespective of the presence of FXa (13 pmol min(-1) 10(-8) platelets). No cleavage by thrombin was observed in either the reconstituted coagulation system or clotting plasma. CONCLUSION: These findings suggest that in vivo the anticoagulant activity of protein S is not down-regulated by FXa or thrombin during activation of coagulation. Our results rather suggest a role for a platelet protease in down regulating the anticoagulant activity of protein S during the hemostatic response. PMID- 16359510 TI - Factor V Leiden improves in vivo hemostasis in murine hemophilia models. AB - The role of factor V Leiden (FVL) as a modifier of the severe hemophilia phenotype is still unclear. We used mice with hemophilia A or B crossed with FVL to elucidate in vivo parameters of hemostasis. Real-time thrombus formation in the microcirculation was monitored by deposition of labeled platelets upon laser induced endothelial injury using widefield microscopy in living animals. No thrombi formed in hemophilic A or B mice following vascular injuries. However, hemophilic mice, either heterozygous or homozygous for FVL, formed clots at all injured sites. Injection of purified activated FV into hemophilic A or B mice could mimic the in vivo effect of FVL. In contrast to these responses to a laser injury in a microvascular bed, FVL did not provide sustained hemostasis following damage of large vessels in a ferric chloride carotid artery injury model, despite of the improvement of clotting times and high circulating thrombin levels. Together these data provide evidence that FVL has the ability to improve the hemophilia A or B phenotype, but this effect is principally evident at the microcirculation level following a particular vascular injury. Our observations may partly explain the heterogeneous clinical evidence of the beneficial role of FVL in hemophilia. PMID- 16359511 TI - Role of coagulation FVIII in septic peritonitis assessed in hemophilic mice. AB - BACKGROUND: Inhibition of blood coagulation appears to be an important therapeutic strategy to improve the outcome in sepsis. However, the beneficial effect of anticoagulant treatment in sepsis is solely based on experimental data using inhibitors of the extrinsic coagulant pathway. The role of the intrinsic pathway of coagulation in the pathogenesis of sepsis has not been explored yet. OBJECTIVE: In the current study, we contribute to determine the role of factor (F)VIII, the key player of the intrinsic coagulant pathway, on host defense against peritonitis. METHOD: Hemizygous FVIII-deficient mice and their wild-type littermates were challenged with 1 x 10(4) bacteria in a septic peritonitis model. RESULTS: The intraperitoneal injection of Escherichia coli led to growth and dissemination of bacteria and provoked an inflammatory response as evident from elevated cytokine levels, increased cell influx into tissues, liver necrosis, and endothelialitis resulting in mortality. The FVIII-deficient genotype slightly reduced bacterial outgrowth but had no effect on markers of inflammation and/or survival. In addition, FVIII-deficient mice showed profound activation of coagulation, thereby improving the hemophilic phenotype of FVIII deficient mice. CONCLUSION: FVIII deficiency slightly modifies host defense in septic peritonitis in mice, but does not influence the final outcome of peritonitis. Therefore, we question the importance of the intrinsic coagulant pathway during sepsis. PMID- 16359512 TI - Modulation of TAFI function through different pathways--implications for the development of TAFI inhibitors. AB - OBJECTIVE: To elucidate the mechanism and the binding regions of monoclonal antibodies (MA) that interfere with thrombin-activatable fibrinolysis inhibitor (TAFI)/activated thrombin-activatable fibrinolysis inhibitor (TAFIa) activity. RESULTS: Of 42 MA, 19 interfere with the TAFI activation/TAFIa activity resulting in an inhibition of up to 92%. Characterization of the mechanism of inhibition revealed that 14 MA blocked the activation of TAFI by thrombin/thrombomodulin completely whereas five MA interfered directly with the enzymatic activity of TAFIa. Surprisingly, the former, except one, induced a significant reduction of clot lysis time whereas the latter did not. Affinity studies using a human/murine TAFI chimer revealed that the binding region of the 14 activation blocking MA is located between AA1 and AA67. MA that inhibit exclusively the activation of TAFI by thrombin/thrombomodulin bind to Gly66. A MA that inhibits the activation of TAFI by both thrombin/thrombomodulin and plasmin binds to Val41. The MA that interfere with the enzymatic activity bind to the TAFIa moiety. CONCLUSIONS: The current study reveals at least three different putative molecular targets in the search for pharmacologically active compounds to modulate TAFIa activity. PMID- 16359513 TI - The phospholipid composition and cholesterol content of platelet-derived microparticles: a comparison with platelet membrane fractions. AB - BACKGROUND: The processes that govern the distribution of molecules between platelets and the microparticles (MP) they release are unknown. Certain proteins are sorted selectively into MP, but lipid sorting has not been studied. OBJECTIVES: To compare the phospholipid composition and cholesterol content of platelet-derived MP obtained with various stimuli with that of isolated platelet membrane fractions. METHODS: Washed platelets from venous blood of healthy individuals (n = 6) were stimulated with collagen, thrombin, collagen plus thrombin, or A23187. Platelet activation, MP release and antigen exposure were assessed by flow cytometry. MPs were isolated by differential centrifugation. Platelet plasma-, granule- and intracellular membranes were isolated from platelet concentrates (n = 3; 10 donors each) by pressure homogenization and Percoll density gradient fractionation. The phospholipid composition and cholesterol content of MPs and membrane fractions were analyzed by high performance thin layer chromatography. RESULTS: The phospholipid composition of MPs was intermediate compared with that of platelet plasma- and granule membranes, and differed significantly from that of intracellular membranes. There were small but significant differences in phospholipid composition between the MPs produced by the various agonists, which paralleled differences in P-selectin exposure in case of the physiological agonists collagen, thrombin, or collagen plus thrombin. The cholesterol content of MPs tended to be higher than that of the three-platelet membrane fractions. CONCLUSIONS: Regarding its phospholipid content, the MP membrane is a composite of the platelet plasma- and granule membranes, showing subtle differences depending on the platelet agonist. The higher cholesterol content of MPs suggests their enrichment in lipid rafts. PMID- 16359514 TI - A 13-bp deletion in alpha(IIb) gene is a founder mutation that predominates in Palestinian-Arab patients with Glanzmann thrombasthenia. AB - Glanzmann thrombasthenia (GT) is a rare autosomal recessive bleeding disorder caused by lack or dysfunction of alpha(IIb)beta3 in platelets. GT is relatively frequent in highly inbred populations. We previously identified a 13-bp deletion in the alpha(IIb) gene that causes in-frame deletion of six amino acids in three Palestinian GT patients. In this study, we determined the molecular basis of GT in all known Palestinian patients, examined whether Jordanian patients harbor the same mutations, analyzed whether there is a founder effect for the 13-bp deletion, and determined the mechanism by which the 13-bp deletion abolishes alpha(IIb)beta3 surface expression. Of 11 unrelated Palestinian patients, eight were homozygous for the 13-bp deletion that displayed common ancestry by haplotype analysis, and was estimated to have occurred 300-600 years ago. Expression studies in baby hamster kidney cells showed that substitution of Cys107 or Trp110 located within the deletion caused defective alpha(IIb)beta3 maturation. Substitution of Trp110, but not of Cys107, prevented fibrinogen binding. The other Palestinian patients harbored three novel mutations: G2374 deletion in alpha(IIb) gene, TT1616-7 deletion in beta3 gene, and IVS14: -3C --> G in beta3 gene. The latter mutation caused cryptic splicing predicting an extended cytoplasmic tail of beta3 and was expressed as dysfunctional alpha(IIb)beta(3). None of 15 unrelated Jordanian patients carried any of the described mutations. PMID- 16359515 TI - Three novel beta-propeller mutations causing Glanzmann thrombasthenia result in production of normally stable pro-alphaIIb, but variably impaired progression of pro-alphaIIbbeta3 from endoplasmic reticulum to Golgi. AB - BACKGROUND: Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by lack of platelet aggregation in response to most physiological agonists and caused by either a lack or dysfunction of the platelet integrin alpha(IIb)beta3 (glycoprotein IIb/IIIa). OBJECTIVES: To determine the molecular basis of GT and characterize the mutations by in vitro expression studies. PATIENTS: We studied three unrelated patients from southern India whose diagnosis was consistent with GT. RESULTS: Immunoprecipitation of the cell lysates and immunoblotting showed no detectable mature alpha(IIb) in the G128S mutant, in contrast to 6% and 33% of the normal amount of mature alpha(IIb) in the S287L and G357S mutants, respectively. Pulse-chase analysis demonstrated pro alpha(IIb) in the mutants comparable with the normal pro-alpha(IIb), but no conversion to mature alpha(IIb) in the G128S mutant, and only trace conversion to mature alpha(IIb) in the S287L and G357S mutants. The disappearance of pro alpha(IIb) in the three mutants was similar to that in cells expressing normal alpha(IIb)beta3 or alpha(IIb) only. All three mutants demonstrated pro alpha(IIb)beta3 complexes and co-localized with an ER marker by immunofluorescence. The G128S mutant showed no co-localization with a Golgi marker, and the other two mutants showed minimal and moderate co-localization with the Golgi marker. CONCLUSIONS: These three beta-propeller mutations do not affect the production of pro-alpha(IIb), its ability to complex with beta3, or its stability, but do cause variable defects in transport of pro-alpha(IIb)beta3 complexes from the endoplasmic reticulum to the Golgi. PMID- 16359516 TI - Persistent production of platelet thromboxane A2 in patients chronically treated with aspirin. AB - BACKGROUND: Patients treated with aspirin may have a reduced sensitivity to its antiplatelet effect. The mechanism accounting for such a reduced sensitivity might involve an impaired interaction of aspirin with cyclooxygenase-1 (COX)-1. OBJECTIVE: We sought to investigate whether platelets from patients under chronic treatment with aspirin still produce TxA2 and whether there is any relationship between the eventual persistent TxA2 formation and platelet aggregation. Finally, whether platelet-derived TxA2 can be inhibited by in vitro addition of aspirin. METHODS: Collagen-induced platelet aggregation and thromboxane-A2 (TxA2) were measured in 196 patients treated with aspirin (100-330 mg day(-1)) because of previous vascular events or presence of risk factors of atherosclerosis. RESULTS: Collagen-induced TxA2 production of the entire cohort was 128.7 +/- 21.6 pg 10( 8) cells, and was significantly correlated with platelet aggregation (Spearman's correlation coefficient = 0.44; P < 0.0001). Patients in the highest quartile of TxA2 showed higher platelet response to collagen (P < 0.0001) when compared with those in the lowest quartile. In a subgroup of 96 patients, platelets were treated in vitro with a TxA2 receptor antagonist (13-azaprostanoic acid) or aspirin before stimulation with collagen. 13-APA acid significantly inhibited platelet aggregation. Aspirin reduced (-72.9%) TxA2 production in patients with TxA2 values above the median but it was ineffective in those with TxA2 values below the median. CONCLUSION: In some patients chronically treated with aspirin platelet production of TxA2 may persist and account for enhanced platelet aggregation. Incomplete inhibition of COX-1 seems to be implicated in persistent TxA2 production. PMID- 16359517 TI - The role of gamma-carboxylation in the anti-apoptotic function of gas6. AB - Gas6 is a novel member of the vitamin K-dependent family of gamma-carboxylated proteins and is a ligand for the receptor tyrosine kinase Axl. Gas6-Axl interactions have been shown to mediate cell survival in vascular endothelium. Although the receptor-binding portion of gas6 lies in the C-terminus, the significance of the N-terminal gamma-carboxylated residues (Gla domain) is not clear. To address this question, this study examines the role of the Gla domain in phospholipid binding as well as in the promotion of cell survival, especially in endothelial cells. The results show that carboxylated gas6 binds to phosphatidylserine-containing phospholipid membranes in an analogous manner to other gamma-carboxylated proteins whereas decarboxylated gas6 does not. The gamma carboxylation inhibitor warfarin abrogates gas6-mediated protection of NIH3T3 fibroblasts from serum starvation-induced apoptosis. Furthermore, the role of gamma-carboxylation in gas6's survival effect on endothelium is demonstrated directly in that only carboxylated, but not decarboxylated, gas6 protects endothelial cells from serum starvation-induced apoptosis. gamma-carboxylation is also required for both Axl phosphorylation and PI3 kinase activation. Taken together, these findings demonstrate that gamma-carboxylation is necessary not only for gas6 binding to phospholipid membranes, but also for gas6-mediated endothelial cell survival. PMID- 16359518 TI - Protease-activated receptors-1 and -2 can mediate endothelial barrier protection: role in factor Xa signaling. AB - Coagulation and inflammation are intimately linked and cellular signaling by coagulation proteases through protease-activated receptors (PARs) may affect pro- and anti-inflammatory responses. Permeability of the endothelial cell barrier at the blood-tissue interface plays a key role in inflammatory disorders such as sepsis. We have recently shown that PAR1 signaling by activated protein C or low concentrations of thrombin can enhance endothelial barrier integrity. In the present study, we analyzed effects of coagulation factor Xa (FXa), which is known to activate both endothelial cell PAR1 and PAR2, on monolayer integrity using a transformed human umbilical vein endothelial cell (HUVEC) line in a dual-chamber system. Preincubation with FXa potently reduced high-dose thrombin-mediated hyperpermeability and basal permeability. FXa was protective at concentrations of 5 nm or higher and proteolytic activity was required. Barrier protective FXa signaling was not affected by cleavage-blocking anti-PAR1 antibodies or by a PAR1 antagonist. Similarly, cleavage-blocking anti-PAR2 alone had no effect, but blocking both PAR1 and PAR2 inhibited barrier protection by FXa. Incubation of the cell layer with a PAR2-specific agonist peptide reduced thrombin-mediated hyperpermeability and basal permeability similar to FXa. In conclusion, not only PAR1, but also PAR2 can mediate barrier protection in endothelial cells and FXa can use either receptor to enhance barrier integrity. Although it is currently unknown whether PAR signaling by FXa has a physiological role, the results suggest a potential protective effect of FXa and other agonists of endothelial PAR2, which should be explored in models of local and systemic inflammation in vivo. PMID- 16359519 TI - Thrombolytic therapy in acute myocardial infarction. PMID- 16359520 TI - Thrombolytic therapy in acute myocardial infarction. PMID- 16359521 TI - Common genomic sequence variation of the prothrombin gene and risk of non-fatal myocardial infarction in white women. PMID- 16359522 TI - The prothrombotic risk of diabetes mellitus in atrial fibrillation and heart failure. PMID- 16359523 TI - Development of antibodies specific to polyanion-modified platelet factor 4 during treatment with fondaparinux. PMID- 16359524 TI - The functional -4C>T polymorphism of the coagulation factor XII gene is not associated with deep venous thrombosis. PMID- 16359525 TI - Low levels of protein Z and the risk of venous thromboembolism. PMID- 16359526 TI - Carboxypeptidase B2 gene polymorphisms and the risk of venous thromboembolism. PMID- 16359527 TI - Rapid single-step detection of the inversion hotspot of mutation in hemophilia A by real-time PCR. PMID- 16359528 TI - Bridging anticoagulation for patients on long-term vitamin-K-antagonists. A prospective 1 year registry of 311 episodes. PMID- 16359529 TI - More on: thrombomodulin gene polymorphisms or haplotypes as potential risk factors for venous thromboembolism: a population-based case-control study. PMID- 16359531 TI - Editorial. PMID- 16359532 TI - The evolving classification of soft tissue tumours: an update based on the new WHO classification. AB - Tumour classifications have become an integral part of modern oncology and, for pathologists, they provide guidelines which facilitate diagnostic and prognostic reproducibility. In many organ systems and most especially over the past decade or so, the World Health Organization (WHO) classifications have become pre eminent, partly enabled by the timely publication of new "blue books" which now incorporate detailed text and copious illustrations. The new WHO classification of soft tissue tumours was introduced in late 2002 and, because it represents a broad consensus view, it has gained widespread acceptance. This review summarizes the changes, both major and minor, which were introduced and briefly describes the significant number of tumour types which have been first recognized or properly characterized during the past decade. Arguably the four most significant conceptual advances have been: (i) the formal recognition that morphologically benign lesions (such as cutaneous fibrous histiocytoma) may very rarely metastasize; (ii) the general acceptance that most pleomorphic sarcomas can be meaningfully subclassified and that so-called malignant fibrous histiocytoma is not a definable entity, but instead represents a wastebasket of undifferentiated pleomorphic sarcomas, accounting for no more than 5% of adult soft tissue sarcomas; (iii) the acknowledgement that most lesions formerly known as haemangiopericytoma show no evidence of pericytic differentiation and, instead, are fibroblastic in nature and form a morphological continuum with solitary fibrous tumour; and (iv) the increasing appreciation that not only do we not know from which cell type(s) most soft tissue tumours originate (histogenesis) but, for many, we do not recognize their line of differentiation or lineage--hence an increasing number of tumours are placed in the "uncertain differentiation" category. PMID- 16359533 TI - The role of genetic testing in soft tissue sarcoma. AB - Soft tissue tumours represent a heterogeneous group of mesenchymal lesions and their classification continues to evolve as a result of incorporating advances in cytogenetic and molecular techniques. In the last decade traditional diagnostic approaches were supplemented with a significant number of reliable molecular diagnostic tools, detecting tumour type-specific genetic alterations. In addition, the successful application of some of these techniques to formalin fixed paraffin-embedded tissue made it possible to subject a broader range of clinical material to molecular analysis. Thus, molecular genetics has already become an integral part of the work-up in some tumours, such as paediatric small blue round cell tumours, which demonstrate characteristic translocations. Several lines of evidence suggest that sarcomas can be divided into two major genetic groups: (i) sarcomas with specific genetic alterations and usually simple karyotypes, such as reciprocal chromosomal translocations (e.g. FUS-DDIT3 in myxoid liposarcoma) and specific oncogenic mutations (e.g. KIT mutation in gastrointestinal stromal tumours); and (i) sarcomas with non-specific genetic alterations and complex unbalanced karyotypes. Some of these genetic abnormalities, including chromosomal numerical changes, translocations, gene amplifications or large deletions can be apparent at the cytogenetic level (karyotyping, fluorescence in situ hybridization), while others, such as small deletions, insertions or point mutations, require molecular genetic techniques (polymerase chain reaction and sequence analysis). This review focuses on the applicability of genetic testing in the diagnosis and prognosis of soft tissue sarcomas, and gives a realistic appraisal of the ancillary role of molecular techniques, including its advantages and limitations. PMID- 16359534 TI - The role of microarray technologies in the study of soft tissue tumours. AB - Array technologies (gene array, tissue microarray and others) are being used in a growing number of research projects involving soft tissue tumours. Gene array techniques allow for measurements of RNA expression levels or gene copy number changes for a large number of genes in a single specimen. A complementary technique, tissue microarrays, allows for the measurement of expression of a single gene in a large number of specimens. These techniques and similar ones have created a fundamentally new approach to the investigation of soft tissue tumours. This review addresses some of the advantages, problems, and solutions to those problems that come with these technologies. PMID- 16359535 TI - The comparative roles of electron microscopy and immunohistochemistry in the diagnosis of soft tissue tumours. AB - Electron microscopy has contributed to the diagnosis of soft tissue tumours for four decades, and immunohistochemistry for two. Because of its relative ease of use and interpretation, the latter technique has become extensively and routinely applied to identify lines of differentiation in benign soft tissue tumours and in sarcomas. The use of electron microscopy has declined but retains a role because few antibodies are wholly specific or fully sensitive, some tumours are polyphenotypic or divergent in differentiation, and others have no specific antigens. Immunohistochemistry is superior in diagnosis of smooth muscle tumours, small round cell tumours, sarcomas with epithelioid morphology, and most synovial sarcomas. Electron microscopy is of particular value for peripheral nerve sheath tumours, marker-negative synovial sarcomas, pleomorphic sarcomas and mesotheliomas. As with all adjunctive techniques, immunohistochemistry and electron microscopy should be used in a complementary fashion according to the nature of the diagnostic problem. PMID- 16359536 TI - Grading of soft tissue sarcomas: the challenge of providing precise information in an imprecise world. AB - By identifying patients at greatest risk for distant metastasis and, hence, most likely to benefit from adjuvant therapy, the grading of sarcomas has been one of the most important contributions pathologists have made to the treatment of sarcomas. Over the years, many grading schemes have been proposed and validated as efficacious. The three-tier system proposed by the French Federation of Cancer Centres is precisely defined, easy to use, and is the most widely employed. However, no system performs perfectly on all sarcomas. Sarcomas that do not lend themselves well to grading include (i) those in which grade provides no incremental information over histological subtypes (e.g. well-differentiated liposarcoma/atypical lipomatous neoplasm, Ewing's sarcoma); (ii) tumours traditionally considered "ungradable" (e.g. epithelioid sarcoma, clear cell sarcoma, angiosarcoma); and (iii) sarcomas that customarily have been graded but in which grade has recently been shown not to correlate well with outcome (e.g. malignant peripheral nerve sheath tumour). Consequently, several sarcoma-specific risk stratification schemes have been proposed. The future may well witness a synthesis of these two approaches. Nomograms, which incorporate clinical, histological and demographic findings, have proved accurate in predicting disease specific survival in sarcomas. Diagnosis and grading are increasingly based on tissue obtained by core needle biopsy, which poses new challenges for pathologists, particularly if neoadjuvant therapy is to be given. Grading on needle biopsies may require a two-tier grading system (i.e. low versus high grade) and a close dialogue with clinicians to resolve ambiguities. PMID- 16359537 TI - Classification of pleomorphic sarcomas: where are we now? AB - Until a decade ago, so-called pleomorphic and storiform malignant fibrous histiocytoma (MFH) represented the most frequently diagnosed sarcoma, accounting for approximately 40% of adult mesenchymal malignancies. However, the latest World Health Organization classification of soft tissue tumours considers MFH a synonym for undifferentiated pleomorphic sarcoma. Historically, the term MFH was introduced in the medical literature in 1963 by Ozzello, O'Brien and Stout, on the basis of the acquisition of phagocytic properties observed in cultured fibroblasts. The existence of MFH as a well-defined clinicopathological entity became rapidly very popular and by the mid 1980s MFH represented the most common sarcoma in adults. With the advent of electron microscopy, immunohistochemistry and molecular genetics, it became clear that the so-called "facultative fibroblast" theory had no scientific grounds and, in 1992, a milestone paper eventually brought attention to the concept that MFH merely represented a morphological pattern shared by a wide variety of poorly differentiated malignant neoplasms, which include specific subtypes of pleomorphic sarcomas. Currently, accurate subclassification of pleomorphic sarcomas is mandatory as it enables recognition of non-sarcomatous lesions as well as pleomorphic neoplasms not associated with aggressive behaviour. Furthermore, as myogenic differentiation predicts aggressive clinical behaviour among pleomorphic sarcomas, precise histotyping allows prognostic stratification of patients. PMID- 16359538 TI - Solitary fibrous tumour and haemangiopericytoma: evolution of a concept. AB - Haemangiopericytoma (HPC) was described in 1942 by Stout and Murray as a distinctive soft tissue neoplasm, presumably of pericytic origin, exhibiting a characteristic well-developed "staghorn" branching vascular pattern. Over the years, it appeared that this growth pattern was a non-specific one, shared by numerous, unrelated benign and malignant lesions, and that HPC was better considered as a diagnosis of exclusion. Three categories of lesion may now be individualized within the heterogeneous group of HPC-like neoplasms. The first category corresponds to those non-HPC neoplasms that occasionally display HPC like features (e.g. synovial sarcoma). Lesions belonging to the second category show clear evidence of myoid/pericytic differentiation and correspond to true HPCs. They generally show a benign clinical course, and include glomangiopericytoma/myopericytoma, infantile myofibromatosis (previously called infantile HPC), and a subset of sinonasal HPCs. The third category is the solitary fibrous tumour (SFT) lesional group, which includes fibrous-to-cellular SFTs, and related lesions such as giant cell angiofibromas and lipomatous HPCs. In practice, any HPC-like lesion can be allocated to one of these categories, leaving the ill-defined "haemangiopericytoma" category empty. PMID- 16359539 TI - PEComa: what do we know so far? AB - PEComas (tumours showing perivascular epithelioid cell differentiation) are a family of related mesenchymal neoplasms that include angiomyolipoma, lymphangiomyomatosis, clear cell "sugar" tumour of the lung, and a group of rare, morphologically and immunophenotypically similar lesions arising at a variety of visceral and soft tissue sites. These tumours all share a distinctive cell type, the perivascular epithelioid cell or "PEC' (which has no known normal tissue counterpart). PEComas show a marked female predominance and are composed of nests and sheets of usually epithelioid but occasionally spindled cells with clear to granular eosinophilic cytoplasm and a focal association with blood vessel walls. PEComas appear to arise most commonly at visceral (especially gastrointestinal and uterine), retroperitoneal, and abdominopelvic sites, with a subset occurring in somatic soft tissue and skin. Nearly all PEComas show immunoreactivity for both melanocytic (HMB-45 and/or melan-A) and smooth muscle (actin and/or desmin) markers. A subset of PEComas behave in a malignant fashion. This review examines the members of the PEComa family, with an emphasis on lesions arising outside of the kidney, lung and liver, and discusses preliminary evidence for pathological features that might predict malignant behaviour. PMID- 16359540 TI - Gastrointestinal stromal tumours: an update. AB - Recently, there has been intense interest in the study of gastrointestinal stromal tumour (GIST); one might call it a virtual GIST revolution. This is due largely to the realization that most GISTs express KIT and harbour activating c KIT (KIT) or platelet-derived growth factor receptor-alpha (PDGFRA) receptor tyrosine kinase mutations that can be targeted by small molecule pharmacological inhibitors. Pathologists have benefited greatly from this revolution, mainly in the form of an improved ability to classify GISTs and, even more recently, in understanding the molecular underpinnings that underlie many fascinating clinical and pathological correlations. It is the purpose of this review to summarize recent developments in GIST classification and the molecular pathogenesis of GIST. PMID- 16359541 TI - Evaluation of biological potential of smooth muscle tumours. AB - Smooth muscle tumours (SMTs) have been traditionally divided into benign leiomyomas (LM) and malignant leiomyosarcomas (LMS) based on cytological atypia, mitotic activity and other criteria. In most instances, this dichotomous approach works, but in some instances the biological potential cannot be determined with certainty. This is often because some, but not all criteria for malignancy have been met or because the tumours are occurring in unusual settings for which there are sparse substantive data. Tumours falling into the latter categories are often designated as 'smooth muscle tumours of uncertain malignant potential'. For most non-hormonally influenced SMTs, the presence of significant atypia plus mitotic activity equates with a diagnosis of LMS. However, not all tumours classified as LMSs have a similar prognosis, as a number of other factors, including tumour size, depth, grade and resectability, affect outcome. For example, cutaneous SMTs, regardless of mitotic activity and atypia, have potential largely limited to local recurrence, whereas subcutaneous and deep LMSs have a definite metastatic potential. Angioleiomyoma is the most common SMT of peripheral soft tissues, but deep peripheral LMs are distinctly rare and should be approached with caution. Hormonally influenced oestrogen- and progesterone receptor-positive uterine and extrauterine SMTs in women have unique criteria, including the allowance of higher mitotic activity for the benign LM designation. SMTs of female genital tract can be assessed with criteria similar to uterine tumours. Because of the rarity of these tumours, experience is more limited, and more caution is needed to assess the potential of tumours with mitotic activity and atypia. This review summarizes the current knowledge, guidelines, prognostic data and controversies for the classification of SMTs of soft tissue and most visceral sites. PMID- 16359542 TI - Postradiation vascular proliferations: an increasing problem. AB - The occurrence of cutaneous vascular lesions is a rare but well-documented complication of radiation treatment and may be associated with significant morbidity as well as mortality. The overall incidence is low but appears to be rising due to a change in the prevailing treatment of breast carcinoma with increased use of radiation in the setting of breast-conserving therapy for stage 1 and 2 disease. The spectrum of postradiation vascular lesions is wide and ranges from atypical vascular lesions with reportedly benign clinical behaviour to frank cutaneous angiosarcoma. There is, however, significant clinical as well as histological overlap. It is frequently difficult to classify these postradiation vascular lesions accurately and they create an emerging diagnostic and therapeutic challenge to both pathologists and clinicians. Experience with these vascular lesions is very limited, and this article aims to provide an overview of our current understanding and concept of radiation-associated vascular lesions with focus on their clinical and histological presentation as well as behaviour and treatment. PMID- 16359543 TI - Sal-Site: integrating new and existing ambystomatid salamander research and informational resources. AB - Salamanders of the genus Ambystoma are a unique model organism system because they enable natural history and biomedical research in the laboratory or field. We developed Sal-Site to integrate new and existing ambystomatid salamander research resources in support of this model system. Sal-Site hosts six important resources: 1) Salamander Genome Project: an information-based web-site describing progress in genome resource development, 2) Ambystoma EST Database: a database of manually edited and analyzed contigs assembled from ESTs that were collected from A. tigrinum tigrinum and A. mexicanum, 3) Ambystoma Gene Collection: a database containing full-length protein-coding sequences, 4) Ambystoma Map and Marker Collection: an image and database resource that shows the location of mapped markers on linkage groups, provides information about markers, and provides integrating links to Ambystoma EST Database and Ambystoma Gene Collection databases, 5) Ambystoma Genetic Stock Center: a website and collection of databases that describe an NSF funded salamander rearing facility that generates and distributes biological materials to researchers and educators throughout the world, and 6) Ambystoma Research Coordination Network: a web-site detailing current research projects and activities involving an international group of researchers. Sal-Site is accessible at http://www.ambystoma.org. PMID- 16359544 TI - Introduction of in vitro transcribed ENO1 mRNA into neuroblastoma cells induces cell death. AB - BACKGROUND: Neuroblastoma is a solid tumour of childhood often with an unfavourable outcome. One common genetic feature in aggressive tumours is 1p deletion. The alpha-enolase (ENO1) gene is located in chromosome region 1p36.2, within the common region of deletion in neuroblastoma. One alternative translated product of the ENO1 gene, known as MBP-1, acts as a negative regulator of the c myc oncogene, making the ENO1 gene a candidate as a tumour suppressor gene. METHODS: Methods used in this study are transfection of cDNA-vectors and in vitro transcribed mRNA, cell growth assay, TUNEL-assay, real-time RT-PCR (TaqMan) for expression studies, genomic sequencing and DHPLC for mutation detection. RESULTS: Here we demonstrate that transfection of ENO1 cDNA into 1p-deleted neuroblastoma cell lines causes' reduced number of viable cells over time compared to a negative control and that it induces apoptosis. Interestingly, a similar but much stronger dose-dependent reduction of cell growth was observed by transfection of in vitro transcribed ENO1 mRNA into neuroblastoma cells. These effects could also be shown in non-neuroblastoma cells (293-cells), indicating ENO1 to have general tumour suppressor activity. Expression of ENO1 is detectable in primary neuroblastomas of all different stages and no difference in the level of expression can be detected between 1p-deleted and 1p-intact tumour samples. Although small numbers (11 primary neuroblastomas), there is some evidence that Stage 4 tumours has a lower level of ENO1-mRNA than Stage 2 tumours (p = 0.01). However, mutation screening of 44 primary neuroblastomas of all different stages, failed to detect any mutations. CONCLUSION: Our studies indicate that ENO1 has tumour suppressor activity and that high level of ENO1 expression has growth inhibitory effects. PMID- 16359545 TI - Dimerization and enzymatic activity of fungal 17beta-hydroxysteroid dehydrogenase from the short-chain dehydrogenase/reductase superfamily. AB - BACKGROUND: 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17beta-HSDcl) is a member of the short-chain dehydrogenase/reductase (SDR) superfamily. SDR proteins usually function as dimers or tetramers and 17beta-HSDcl is also a homodimer under native conditions. RESULTS: We have investigated here which secondary structure elements are involved in the dimerization of 17beta-HSDcl and examined the importance of dimerization for the enzyme activity. Sequence similarity with trihydroxynaphthalene reductase from Magnaporthe grisea indicated that Arg129 and His111 from the alphaE-helices interact with the Asp121, Glu117 and Asp187 residues from the alphaE and alphaF helices of the neighbouring subunit. The Arg129Asp and His111Leu mutations both rendered 17beta-HSDcl monomeric, while the mutant 17beta-HSDcl-His111Ala was dimeric. Circular dichroism spectroscopy analysis confirmed the conservation of the secondary structure in both monomers. The three mutant proteins all bound coenzyme, as shown by fluorescence quenching in the presence of NADP+, but both monomers showed no enzymatic activity. CONCLUSION: We have shown by site-directed mutagenesis and structure/function analysis that 17beta-HSDcl dimerization involves the alphaE and alphaF helices of both subunits. Neighbouring subunits are connected through hydrophobic interactions, H-bonds and salt bridges involving amino acid residues His111 and Arg129. Since the substitutions of these two amino acid residues lead to inactive monomers with conserved secondary structure, we suggest dimerization is a prerequisite for catalysis. A detailed understanding of this dimerization could lead to the development of compounds that will specifically prevent dimerization, thereby serving as a new type of inhibitor. PMID- 16359547 TI - Mutations in the mitochondrial DNA D-loop region are frequent in cervical cancer. AB - BACKGROUND: Mitochondrial DNA (mtDNA) is known for high mutation rates caused by lack of protective histones, inefficient DNA repair systems, and continuous exposure to mutagenic effects of oxygen radicals. Alterations in the non-coding displacement (D) loop of mitochondrial DNA are present in many cancers. It has been suggested that the extent of mitochondrial DNA mutations might be useful in the prognosis of cancer outcome and/or the response to certain therapies. In order to investigate whether a high incidence of mutations exist in mitochondrial DNA of cervical cancer patients, we examined the frequency of mutations in the D loop region in 19 patients of cervical cancer. RESULTS: Mutations, often multiple, were detected in 18 of 19 (95%) patients. The presence of mutations correlated with Human Papilloma Virus (HPV) infection in these patients. Mutations were also detected in normal samples and lymphocytes obtained from cervical cancer patients, but their frequency of occurrence was much lower as compared to the cervical cancer tissues. CONCLUSION: Our findings indicate that D loop alterations are frequent in cervical cancers and are possibly caused by HPV infection. There was no association of mtDNA D-loop mutations with the histopathological grade and tumor staging. PMID- 16359546 TI - Cigarette smoke-induced pulmonary emphysema in scid-mice. Is the acquired immune system required? AB - BACKGROUND: Chronic obstructive pulmonary disease is associated with a chronic inflammatory response of the host to chronic exposure to inhaled toxic gases and particles. Although inflammatory cells of both the innate and adaptive immune system infiltrate the lungs in pulmonary emphysema and form lymphoid follicles around the small airways, the exact role of the acquired immune system in the pathogenesis of emphysema is not known. METHODS: In this study, wild type Balb/c mice and immunodeficient scid mice--which lack functional B- and T-cells--were exposed to mainstream cigarette smoke (CS) for 5 weeks or 6 months. RESULTS: Subacute CS-exposure for 5 weeks significantly increased innate inflammatory cells (neutrophils, macrophages and dendritic cells) in the bronchoalveolar lavage (BAL) fluid of wild type mice and scid mice, which correlated with the CS induced upregulation of the chemokines Monocyte Chemotactic Protein-1, Macrophage Inflammatory Protein-3alpha and KC (= mouse Interleukin-8). Chronic CS-exposure for 6 months significantly increased the number of neutrophils, macrophages, dendritic cells, CD4+ and CD8+ T-lymphocytes in BAL fluid and lungs of wild type mice compared to air-exposed littermates, and augmented the size and number of peribronchial lymphoid follicles. In contrast, neither B-lymphocytes, nor T lymphocytes, nor lymphoid follicles could be discerned in the lungs of air- or CS exposed scid mice. Importantly, chronic CS-exposure induced pulmonary emphysema in both wild type animals and scid mice, as evidenced by a significant increase in the mean linear intercept and the destructive index of CS-exposed versus air exposed animals. The CS-induced emphysema was associated with increased mRNA expression of matrix metalloproteinase-12 in the lungs and increased protein levels of Tumor Necrosis Factor-alpha in the BAL fluid of CS-exposed Balb/c and scid mice compared to air-exposed littermates. CONCLUSION: This study suggests that the adaptive immune system is not required per se to develop pulmonary emphysema in response to chronic CS-exposure, since emphysema can be induced in scid mice, which lack lymphoid follicles as well as functional B- and T-cells. PMID- 16359548 TI - Impact of efalizumab on patient-reported outcomes in high-need psoriasis patients: results of the international, randomized, placebo-controlled Phase III Clinical Experience Acquired with Raptiva (CLEAR) trial [NCT00256139]. AB - BACKGROUND: Chronic psoriasis can negatively affect patients' lives. Assessing the impact of treatment on different aspects of a patient's health-related quality of life (HRQOL) is therefore important and relevant in trials of anti psoriasis agents. The recombinant humanized IgG1 monoclonal antibody efalizumab targets multiple T-cell-dependent steps in the immunopathogenesis of psoriasis. Efalizumab has demonstrated safety and efficacy in several clinical trials, and improves patients' quality of life. OBJECTIVE: To evaluate the impact of efalizumab on HRQOL and other patient-reported outcomes in patients with moderate to severe plaque psoriasis, including a large cohort of High-Need patients for whom at least 2 other systemic therapies were unsuitable because of lack of efficacy, intolerance, or contraindication. METHODS: A total of 793 patients were randomized in a 2:1 ratio to receive efalizumab 1 mg/kg/wk (n = 529) or placebo (n = 264) for 12 weeks. The study population included 526 High-Need patients (342 efalizumab, 184 placebo). The treatment was evaluated by patients using the HRQOL assessment tools Short Form-36 (SF-36) and Dermatology Life Quality Index (DLQI). Other patient-reported assessments included the Psoriasis Symptom Assessment (PSA), a visual analog scale (VAS) for itching, and the Patient's Global Psoriasis Assessment (PGPA). RESULTS: Efalizumab was associated with improvements at Week 12 from baseline in patient-reported outcomes, both in the total study population and in the High-Need cohort. Among all efalizumab-treated patients, the DLQI improved by 5.7 points from baseline to Week 12, relative to an improvement of 2.3 points for placebo patients (P < .001). Corresponding improvements in DLQI in the High-Need cohort were 5.4 points for efalizumab compared to 2.3 for placebo (P < .001). Improvements from baseline on the SF-36, PSA, PGPA, and itching VAS at Week 12 were also significantly greater in efalizumab-treated patients than for placebo. CONCLUSION: A 12-week course of efalizumab improved HRQOL and other patient-reported outcomes in patients with moderate to severe plaque psoriasis. The benefits of efalizumab therapy in High Need patients were similar to those observed in the total study population, indicating that the beneficial impact of efalizumab on QOL is consistent regardless of disease severity, prior therapy, or contraindications to previous therapies. PMID- 16359549 TI - It's the virus, stupid--part 2. AB - This editorial presents Retrovirology's choice for the best basic science "retrovirus paper of the year 2005". PMID- 16359550 TI - Induction and regulation of matrix metalloproteinase-12 in human airway smooth muscle cells. AB - BACKGROUND: The elastolytic enzyme matrix metalloproteinase (MMP)-12 has been implicated in the development of airway inflammation and remodeling. We investigated whether human airway smooth muscle cells could express and secrete MMP-12, thereby participating in the pathogenesis of airway inflammatory diseases. METHODS: Laser capture microdissection was used to collect smooth muscle cells from human bronchial biopsy sections. MMP-12 mRNA expression was analysed by quantitative real-time RT-PCR. MMP-12 protein expression and secretion from cultured primary airway smooth muscle cells was further analysed by Western blot. MMP-12 protein localization in bronchial tissue sections was detected by immunohistochemistry. MMP-12 activity was determined by zymography. The TransAM AP-1 family kit was used to measure c-Jun activation and nuclear binding. Analysis of variance was used to determine statistical significance. RESULTS: We provide evidence that MMP-12 mRNA and protein are expressed by in situ human airway smooth muscle cells obtained from bronchial biopsies of normal volunteers, and of patients with asthma, COPD and chronic cough. The pro inflammatory cytokine, interleukin (IL)-1beta, induced a >100-fold increase in MMP-12 gene expression and a >10-fold enhancement in MMP-12 activity of primary airway smooth muscle cell cultures. Selective inhibitors of extracellular signal regulated kinase, c-Jun N-terminal kinase and phosphatidylinositol 3-kinase reduced the activity of IL-1beta on MMP-12, indicating a role for these kinases in IL-1beta-induced induction and release of MMP-12. IL-1beta-induced MMP-12 activity and gene expression was down-regulated by the corticosteroid dexamethasone but up-regulated by the inflammatory cytokine tumour necrosis factor (TNF)-alpha through enhancing activator protein-1 activation by IL-1beta. Transforming growth factor-beta had no significant effect on MMP-12 induction. CONCLUSION: Our findings indicate that human airway smooth muscle cells express and secrete MMP-12 that is up-regulated by IL-1beta and TNF-alpha. Bronchial smooth muscle cells may be an important source of elastolytic activity, thereby participating in remodeling in airway diseases such as COPD and chronic asthma. PMID- 16359551 TI - The effects of a low-carbohydrate, ketogenic diet on the polycystic ovary syndrome: a pilot study. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder affecting women of reproductive age and is associated with obesity, hyperinsulinemia, and insulin resistance. Because low carbohydrate diets have been shown to reduce insulin resistance, this pilot study investigated the six month metabolic and endocrine effects of a low-carbohydrate, ketogenic diet (LCKD) on overweight and obese women with PCOS. RESULTS: Eleven women with a body mass index >27 kg/m2 and a clinical diagnosis of PCOS were recruited from the community. They were instructed to limit their carbohydrate intake to 20 grams or less per day for 24 weeks. Participants returned every two weeks to an outpatient research clinic for measurements and reinforcement of dietary instruction. In the 5 women who completed the study, there were significant reductions from baseline to 24 weeks in body weight (-12%), percent free testosterone (-22%), LH/FSH ratio (-36%), and fasting insulin (-54%). There were non-significant decreases in insulin, glucose, testosterone, HgbA1c, triglyceride, and perceived body hair. Two women became pregnant despite previous infertility problems. CONCLUSION: In this pilot study, a LCKD led to significant improvement in weight, percent free testosterone, LH/FSH ratio, and fasting insulin in women with obesity and PCOS over a 24 week period. PMID- 16359552 TI - A novel mass spectrometry-based assay for GSK-3beta activity. AB - BACKGROUND: As a component of the progression from genomic to proteomic analysis, there is a need for accurate assessment of protein post-translational modifications such as phosphorylation. Traditional kinase assays rely heavily on the incorporation of gamma-P32 radiolabeled isotopes, monoclonal anti-phospho protein antibodies, or gel shift analysis of substrate proteins. In addition to the expensive and time consuming nature of these methods, the use of radio ligands imposes restrictions based on the half-life of the radionucleotides and pose potential health risks to researchers. With the shortcomings of traditional assays in mind, the aim of this study was to develop a high throughput, non radioactive kinase assay for screening Glycogen Synthase Kinase-3beta (GSK-3beta) activity. RESULTS: Synthetic peptide substrates designed with a GSK-3beta phosphorylation site were assayed with both recombinant enzyme and GSK-3beta immunoprecipitated from NIH 3T3 fibroblasts. A molecular weight shift equal to that of a single phosphate group (80 Da.) was detected by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) in a GSK 3beta target peptide (2B-Sp). Not only was there a dose-dependent response in molecular weight shift to the amount of recombinant GSK-3beta used in this assay, this shift was also inhibited by lithium chloride (LiCl), in a dose-dependent manner. CONCLUSION: We present here a novel method to sensitively measure peptide phosphorylation by GSK-3beta that, due to the incorporation of substrate controls, is applicable to either purified enzyme or cell extracts. Future studies using this method have the potential to elucidate the activity of GSK 3beta in vivo, and to screen enzyme activity in relation to a variety of GSK 3beta related disorders. PMID- 16359554 TI - The influence of objectives, learning experiences and examination blueprint on medical students' examination preparation. AB - BACKGROUND: The influence of intended and informal curricula on examination preparation has not been extensively studied. This study aims to firstly describe how students utilized components of intended and informal curricula to guide examination preparation, and secondly to study the relationship between examination preparation and performance. METHODS: Students received a pre examination questionnaire to identify components from the intended curriculum (objectives and examination blueprint), and informal curriculum (content emphasised during lectures and small groups), used during examination preparation. Multiple logistic regression was used to study the relationship between these variables and student performance (above versus at or below average). RESULTS: Eighty-one students participated. There was no difference in the proportions using the examination blueprint, content emphasised during lectures, and content emphasised during small groups (87-93%) but fewer students used objectives (35%, p < 0.001). Objectives use was associated with reduced odds of above average examination performance (adjusted odds ratio 0.27 [0.07, 0.97], p = 0.04). CONCLUSION: When preparing for the renal course examination, students were influenced at least as much by the informal as the intended curriculum. Of the two intended curriculum components, the examination blueprint appeared to be more widely used than the course objectives. This decreased use of objectives on examination preparation did not appear to have a detrimental effect on student performance. PMID- 16359555 TI - Hemangiopericytoma of the spleen: unusual presentation as multiple abscess. AB - BACKGROUND: Hemangiopericytoma is a soft tissue vascular neoplasm arising from capillary pericytes and is found throughout the body in soft tissues and bone. It was first described in 1942. Primary vascular neoplasm of the spleen constitutes the majority of nonhaematolymphoid splenic tumors like haemangioma, lymphangioma, hemangioendothelioma, hemangiopericytoma etc. Splenic hemangiopericytoma is a rare tumor and probably first case was described in 1989. Uptill now only eight cases are reported in the English literature. CASE PRESENTATION: A-35-year old male presented with fever and dull aching pain in left hypochondriac region. Radiological evaluation showed presence of multiple abscesses in spleen. Investigations were done to rule out common causes of abscess in spleen. After failure of medical management, he was subjected to elective splenectomy. There were dense adhesions between the spleen and the adjacent structures and the diaphragm. The histopathology of the resected specimen showed hemangiopericytoma of spleen. CONCLUSION: The present case illustrate that the hemangiopericytoma of spleen can mimic as multiple abscess. Splenectomy is the treatment of choice. PMID- 16359553 TI - Plasmodium yoelii 17XL infection up-regulates RANTES, CCR1, CCR3 and CCR5 expression, and induces ultrastructural changes in the cerebellum. AB - BACKGROUND: Malaria afflicts 300-500 million people causing over 1 million deaths globally per year. The immunopathogenesis of malaria is mediated partly by co mplex cellular and immunomodulator interactions involving co-regulators such as cytokines and adhesion molecules. However, the role of chemokines and their receptors in malaria immunopathology remains unclear. RANTES (Regulated on Activation Normal T-Cell Expressed and Secreted) is a chemokine involved in the generation of inflammatory infiltrates. Recent studies indicate that the degradation of cell-cell junctions, blood-brain barrier dysfunction, recruitment of leukocytes and Plasmodium-infected erythrocytes into and occlusion of microvessels relevant to malaria pathogenesis are associated with RANTES expression. Additionally, activated lymphocytes, platelets and endothelial cells release large quantities of RANTES, thus suggesting a unique role for RANTES in the generation and maintenance of the malaria-induced inflammatory response. The hypothesis of this study is that RANTES and its corresponding receptors (CCR1, CCR3 and CCR5) modulate malaria immunopathogenesis. A murine malaria model was utilized to evaluate the role of this chemokine and its receptors in malaria. METHODS: The alterations in immunomodulator gene expression in brains of Plasmodium yoelii 17XL-infected mice was analysed using cDNA microarray screening, followed by a temporal comparison of mRNA and protein expression of RANTES and its corresponding receptors by qRT-PCR and Western blot analysis, respectively. Plasma RANTES levels was determined by ELISA and ultrastructural studies of brain sections from infected and uninfected mice was conducted. RESULTS: RANTES (p < 0.002), CCR1 (p < 0.036), CCR3 (p < 0.033), and CCR5 (p < 0.026) mRNA were significantly upregulated at peak parasitaemia and remained high thereafter in the experimental mouse model. RANTES protein in the brain of infected mice was upregulated (p < 0.034) compared with controls. RANTES plasma levels were significantly upregulated; two to three fold in infected mice compared with controls (p < 0.026). Some distal microvascular endothelium in infected cerebellum appeared degraded, but remained intact in controls. CONCLUSION: The upregulation of RANTES, CCR1, CCR3, and CCR5 mRNA, and RANTES protein mediate inflammation and cellular degradation in the cerebellum during P. yoelii 17XL malaria. PMID- 16359556 TI - Dimensions of the posterior fossa in patients symptomatic for Chiari I malformation but without cerebellar tonsillar descent. AB - BACKGROUND: Chiari I malformation (CMI) is diagnosed by rigid radiographic criteria along with appropriate clinical symptomatology. The aim of this study was to investigate the dimensions of the posterior cranial fossa in patients without significant tonsillar descent but with symptoms comparable to CMI. METHODS: Twenty-two patients with signs and symptoms comparable to CMI but without accepted radiographic criteria of tonsillar descent > 3-5 mm were referred to our clinic for evaluation. A history and physical examination were performed on all patients. In reviewing their MRI scans, nine morphometric measurements were recorded. The measurements were compared to measurements from a cohort of twenty-five individuals with cranial neuralgias from our practice. RESULTS: For patients with Chiari-like symptomatology, the following statistically significant abnormalities were identified: reduced length of the clivus, reduced length of basisphenoid, reduced length of basiocciput, and increased angle of the tentorium. Multiple morphometric studies have demonstrated similar findings in CMI. CONCLUSION: The current classification of CMI is likely too restrictive. Preliminary morphologic data suggests that a subgroup of patients exists with tonsillar descent less than 3 mm below the foramen magnum but with congenitally hypoplastic posterior fossa causing symptomatology consistent with CMI. PMID- 16359557 TI - Remunerating private psychiatrists for participating in case conferences. AB - BACKGROUND: On 1 November 2000, a series of new item numbers was added to the Medicare Benefits Schedule, which allowed for case conferences between physicians (including psychiatrists) and other multidisciplinary providers. On 1 November 2002, an additional set of numbers was added, designed especially for use by psychiatrists. This paper reports the findings of an evaluation of these item numbers. RESULTS: The uptake of the item numbers in the three years post their introduction was low to moderate at best. Eighty nine psychiatrists rendered 479 case conferences at a cost to the Health Insurance Commission of $70,584. Psychiatrists who have used the item numbers are generally positive about them, as are consumers. Psychiatrists who have not used them have generally not done so because of a lack of knowledge, rather than direct opposition. The use of the item numbers is increasing over time, perhaps as psychiatrists become more aware of their existence and of their utility in maximising quality of care. CONCLUSION: The case conferencing item numbers have potential, but as yet this potential is not being realised. Some small changes to the conditions associated with the use of the item numbers could assist their uptake. PMID- 16359558 TI - Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants. AB - BACKGROUND: We describe novel plasmid vectors for transient gene expression using Agrobacterium, infiltrated into Nicotiana benthamiana leaves. We have generated a series of pGreenII cloning vectors that are ideally suited to transient gene expression, by removing elements of conventional binary vectors necessary for stable transformation such as transformation selection genes. RESULTS: We give an example of expression of heme-thiolate P450 to demonstrate effectiveness of this system. We have also designed vectors that take advantage of a dual luciferase assay system to analyse promoter sequences or post-transcriptional regulation of gene expression. We have demonstrated their utility by co-expression of putative transcription factors and the promoter sequence of potential target genes and show how orthologous promoter sequences respond to these genes. Finally, we have constructed a vector that has allowed us to investigate design features of hairpin constructs related to their ability to initiate RNA silencing, and have used these tools to study cis-regulatory effect of intron-containing gene constructs. CONCLUSION: In developing a series of vectors ideally suited to transient expression analysis we have provided a resource that further advances the application of this technology. These minimal vectors are ideally suited to conventional cloning methods and we have used them to demonstrate their flexibility to investigate enzyme activity, transcription regulation and post transcriptional regulatory processes in transient assays. PMID- 16359559 TI - Design and implementation of GRIP: a computerized glucose control system at a surgical intensive care unit. AB - BACKGROUND: Tight glucose control by intensive insulin therapy has become a key part of critical care and is an important field of study in acute coronary care. A balance has to be found between frequency of measurements and the risk of hypoglycemia. Current nurse-driven protocols are paper-based and, therefore, rely on simple rules. For safety and efficiency a computer decision support system that employs complex logic may be superior to paper protocols. METHODS: We designed and implemented GRIP, a stand-alone Java computer program. Our implementation of GRIP will be released as free software. Blood glucose values measured by a point-of-care analyzer were automatically retrieved from the central laboratory database. Additional clinical information was asked from the nurse and the program subsequently advised a new insulin pump rate and glucose sampling interval. RESULTS: Implementation of the computer program was uneventful and successful. GRIP treated 179 patients for a total of 957 patient-days. Severe hypoglycemia (< 2.2 mmol/L) only occurred once due to human error. With a median (IQR) of 4.9 (4.2-6.2) glucose measurements per day the median percentage of time in which glucose fell in the target range was 78%. Nurses rated the program as easy to work with and as an improvement over the preceding paper protocol. They reported no increase in time spent on glucose control. CONCLUSION: A computer driven protocol is a safe and effective means of glucose control at a surgical ICU. Future improvements in the recommendation algorithm may further improve safety and efficiency. PMID- 16359561 TI - Memorable medical mentors: XIV Howard C. Taylor, Jr. (1900-1985). PMID- 16359560 TI - A procedure for localisation and electrophysiological characterisation of ion channels heterologously expressed in a plant context. AB - BACKGROUND: In silico analyses based on sequence similarities with animal channels have identified a large number of plant genes likely to encode ion channels. The attempts made to characterise such putative plant channels at the functional level have most often relied on electrophysiological analyses in classical expression systems, such as Xenopus oocytes or mammalian cells. In a number of cases, these expression systems have failed so far to provide functional data and one can speculate that using a plant expression system instead of an animal one might provide a more efficient way towards functional characterisation of plant channels, and a more realistic context to investigate regulation of plant channels. RESULTS: With the aim of developing a plant expression system readily amenable to electrophysiological analyses, we optimised experimental conditions for preparation and transformation of tobacco mesophyll protoplasts and engineered expression plasmids, that were designed to allow subcellular localisation and functional characterisation of ion channels eventually in presence of their putative (possibly over-expressed) regulatory partners. Two inward K+ channels from the Shaker family were functionally expressed in this system: not only the compliant KAT1 but also the recalcitrant AKT1 channel, which remains electrically silent when expressed in Xenopus oocytes or in mammalian cells. CONCLUSION: The level of endogenous currents in control protoplasts seems compatible with the use of the described experimental procedures for the characterisation of plant ion channels, by studying for instance their subcellular localisation, functional properties, structure function relationships, interacting partners and regulation, very likely in a more realistic context than the classically used animal systems. PMID- 16359563 TI - The role of placental oxidative stress and lipid peroxidation in preeclampsia. AB - Preeclampsia is a complex multisystem disorder exclusively seen in human species that is characterized by hypertension and proteinuria. This disorder has the highest maternal and fetal morbidity and mortality of all pregnancy-related complications. Growing evidence suggests that placental oxidative stress is involved in the etiopathogenesis of preeclampsia. Reduced perfusion as a result of abnormal placentation leads to ischemia reperfusion injury to the placenta. Placental oxidative stress, which results from the ischemia reperfusion injury, is being increasingly reported to be involved in the etiopathogenesis of preeclampsia. It has been proposed as a promoter of lipid peroxidation and the endothelial cell dysfunction that is commonly seen in this condition. Although preeclampsia is characterized by increased lipid peroxidation and diminished antioxidant capacity, there is no consensus regarding causality of lipid peroxidation in preeclampsia. In this article, we address the question of the biologic association of lipid peroxidation and preeclampsia. Lipid peroxidation and leukocyte activation may play a pivotal role in endothelial cell dysfunction. We also review the different factors that have been proposed to cause endothelial cell dysfunction in preeclampsia, trials investigating the role of antioxidant supplementation in preeclampsia, and the lack of consensus among the trials. Additional longitudinal studies are necessary to determine if the various oxidative stress biomarkers estimated early in pregnancy can be narrowed to a single marker for predicting preeclampsia. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to recall that placental oxidative stress is involved in the etiopathogenesis of preeclampsia, state that placental oxidative stress results from ischemic reperfusion injury, and explain that ischemic reperfusion injury is a promoter of lipid peroxidation and endothelial cell dysfunction seen in preeclampsia. PMID- 16359564 TI - Future perspectives in the medical treatment of endometriosis. AB - In the last few years, our understanding of the pathogenesis of endometriosis at the cellular and molecular levels has improved significantly. This may give us the opportunity to use new, specific agents for the treatment of this disorder. Despite the effectiveness of the available treatments, novel therapeutic strategies may improve our ability to eliminate endometriotic lesions when present and to prevent the recurrence of endometriosis after surgical treatment. This review focuses on the new, experimental approaches to the medical treatment of endometriosis and its symptoms. The blockage of aromatase activity in endometriotic lesions with an aromatase inhibitor may represent a new step in the medical treatment of endometriosis. Preliminary clinical studies have demonstrated the efficacy of third-generation nonsteroidal aromatase inhibitors (ie, anastrozole and letrozole) in reducing the intensity of pain symptoms associated with the presence of endometriosis. The new selective progesterone receptor modulators may represent a valid hormonal treatment option. Therapeutic manipulation of the immune system through TNFalpha inhibitors may be beneficial in women with endometriosis. New pharmaceutical agents affecting inflammation, angiogenesis, and matrix metalloproteinase activity may prevent or inhibit the development of endometriosis. Further clinical trials may determine if these new therapies are superior to current medical treatment strategies for endometriosis. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians Learning. OBJECTIVES: After completion of this article, the reader should be able to describe the new experimental medical treatments of endometriosis, state that the clinical use of nonsteroidal aromatase inhibitors for endometriosis appears to be efficacious but is based on preliminary clinical data, and recall that the drugs used for endometriosis in the future may include manipulation of the immune system. PMID- 16359565 TI - Massive blood loss and transfusion in obstetrics and gynecology. AB - Massive perioperative or periparturitional bleeding occasionally occurs in obstetric and gynecologic patients. Placenta previa, uterine atony, and ectopic pregnancy are just a few examples of many conditions that could predispose patients to significant blood loss. Therefore, it is important for physicians specializing in obstetrics and gynecology to be proficient in managing episodes of massive hemorrhage and the practice of the most commonly used blood components. We review and update the management of massive hemorrhage for obstetrics and gynecologic patients. In addition, we explore blood component therapy, its risks and benefits. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians Learning. OBJECTIVES: After completion of this article, the reader should be able to explain the necessity of being proficient in managing episodes of massive hemorrhage, list the indications for use of various blood components, and summarize the risks and benefits of blood component therapy. PMID- 16359568 TI - Fracture prevention in postmenopausal osteoporosis: a review of treatment options. AB - Several treatment options are available to reduce the risk of fractures in postmenopausal women with or at risk for osteoporosis. A MEDLINE search was conducted to evaluate anti-fracture and adverse event data of osteoporosis therapies from trials in postmenopausal women. Among the anti-resorptive therapies, the bisphosphonates alendronate and risedronate have demonstrated consistent efficacy in reducing vertebral and nonvertebral fracture risk. Once weekly alendronate and risedronate produced similar improvements in bone mineral density compared with their once-daily counterparts with similar tolerability. Daily injections of teriparatide resulted in statistically significant reductions in the risk of vertebral and nonvertebral fractures, and trials of ibandronate, raloxifene, and calcitonin nasal spray showed reductions in vertebral fracture risk. Hormone therapy has demonstrated clinical fracture risk reduction; however, safety outcomes from the Women's Health Initiative study have raised concerns regarding long-term use of these preparations. These data can guide clinical decision-making regarding the selection of an osteoporosis therapy. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to summarize adverse events data of osteoporosis therapies from trials in postmenopausal women, explain that only a few therapies have shown a consistent efficacy in reducing vertebral and nonvertebral fractures, and state that data from the Women's Health Initiative study have raised concerns regarding long-term use of estrogen progestin therapy. PMID- 16359569 TI - Clinical approach to mental health issues among obstetrician-gynecologists: a review. AB - In recent years, obstetrician-gynecologists have taken on a greater role in the provision of primary care. Mental health has been a topic worthy of further exploration as a result of the high prevalence rates of women presenting in gynecologic settings with depressive, anxiety, or eating-disordered symptoms. The detrimental effects of psychopathology have been well documented in the literature, especially if present during pregnancy. This article provides a review of the literature in the area of clinical practice related to mental health among obstetrician-gynecologists based on searches of the Psyc Info and MEDLINE databases. Lack of recognition and underdiagnosis are common problems that need to be addressed by focused educational initiatives. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to recall the importance of screening for various types of mental disease during an ob/gyn visit; describe the detrimental effects of psychopathology, especially during pregnancy; and explain the importance of educational initiatives in detecting and treating mental disorders. PMID- 16359570 TI - Anticoagulants in pregnancy. AB - Pregnancy is a hypercoagulable state that increases the risk of thromboembolic events. These risks may be further increased in the presence of an acquired or inherited thrombophilia. Thrombophilias have been associated with both maternal and fetal complications. The use of anticoagulants during pregnancy may reduce the risk of maternal thromboses as well as the risk of adverse pregnancy outcomes. The choice of an anticoagulant requires consideration of maternal risks, potential for teratogenicity, the underlying condition necessitating the treatment, and cost. This review examines the options for anticoagulation, the clinical situations that may warrant such treatment, and factors to be considered at delivery and during the postpartum period. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to describe the roles of acquired and inherited thrombophilia in furthering the hypercoagulable state of pregnancy, identify the potential consequences of using anticoagulants during pregnancy, and summarize the treatment options when anticoagulation is required during pregnancy. PMID- 16359575 TI - Extranasopharyngeal angiofibroma of the cheek. AB - Angiofibromas rarely localize in extranasopharyngeal sites. The most common site for extranasopharyngeal angiofibromas is the maxillary sinus. The ethmoid and sphenoid sinuses, nasal septum, middle and inferior turbinates, conjunctiva, molar and retromolar region, and larynx are other sites where extranasopharyngeal angiofibromas have been reported. Only one case of buccal extranasopharyngeal angiofibroma has been reported to date. We present a case of buccal extranasopharyngeal angiofibroma that was excised completely following embolization and we also review the literature. PMID- 16359576 TI - The combined olfactory test in a Chinese population. AB - OBJECTIVE: To establish an olfactory test in the Hong Kong Chinese population. STUDY DESIGN: Prospective analysis of the results of a combined olfactory test (COT) for patients with olfactory dysfunction and for normal subjects attending a secondary and tertiary ear, nose and throat centre. METHOD: Our COT was based on and modified from the Connecticut Chemosensory Research Centre test. It consisted of an odour identification test involving nine substances and a threshold test using a series of threefold dilutions of 1-butanol. From September 1998 to June 2002, 188 Hong Kong Chinese patients with olfactory dysfunction and subjects with normal olfaction were prospectively recruited. The subjects were categorized into normal, hyposmic and anosmic groups. The olfactory function of the main group of subjects (n = 153) was assessed with the COT by a blinded observer and also quantified with a visual analogue scale; the results were analysed to validate the discriminative ability of the COT. Another subgroup of subjects (n = 35) was tested with the COT twice with a two-week interval to evaluate test-retest reliability. RESULTS: There were 42 normal, 68 hyposmic and 43 anosmic subjects within the main group (total n = 153). The identification scores, threshold scores and combined olfactory scores were statistically significantly different between the anosmic, hyposmic and normal groups of subjects (p < 0.001). The combined olfactory score correlated with the visual analogue score and the correlation coefficient was 0.56 (p < 0.01). The mean time spent on each COT was 8.6 minutes (standard deviation = 3.4 minutes). The test-retest reliability of the COT was satisfactory, with a one-way model intraclass correlation coefficient of 0.87 (n = 35). CONCLUSION: The satisfactory discriminative ability and test retest reliability of the COT have been demonstrated in this study. The COT is a feasible method for assessing sense of smell in the Hong Kong Chinese population. PMID- 16359577 TI - Methylene blue toxicity following infusion to localize parathyroid adenoma. AB - The parathyroid glands are small, inconspicuous, and variable in number, colour and position. Their identification is vital for excision of hyper-functioning glands and for preservation of normally functioning ones in patients undergoing thyroidectomy. Intravenous infusion of methylene blue at a dose of 7.5 mg/kg is commonly used to aid visualization of the parathyroid glands intra-operatively. Methylene blue is generally considered benign, and there are only two cases published in the literature reporting toxicity following intravenous infusion- such toxicity is a diagnosis of exclusion. We report a case of methylene blue toxicity resulting in expressive aphasia, confusion and disinhibition following infusion for parathyroid adenoma localization. The patient made a complete recovery over 48 hours. Methaemoglobinaemia was excluded as a cause. We suggest that the mechanism of toxicity was a direct effect of methylene blue, although an adverse interaction with serotonin re-uptake inhibitors could not be excluded. In keeping with the UK National Poisons Information Service recommendations, we have altered our practice and now use methylene blue at a dose not exceeding 4 mg/kg. This has not affected our success rate for identification of parathyroid glands. We report this case to highlight the rare occurrence of methylene blue toxicity when used at a dose of 7.5 mg/kg. PMID- 16359578 TI - Driving and otolaryngology: do we know the rules? AB - Certain medical conditions may affect one's ability to drive safely. In the United Kingdom, the Driver and Vehicle Licensing Agency (DVLA) issues guidelines outlining which medical conditions drivers must notify them of. In order to assess our knowledge of, and ability to apply, the current DVLA guidelines with regard to otolaryngological conditions, a postal survey was sent to all otolaryngologists in Scotland. The responses obtained suggest that the current DVLA guidelines are not well understood or easily applied, and it is recommended that all otolaryngologists familiarize themselves with these guidelines in order to better advise their patients with regard to driving. PMID- 16359579 TI - Mucosal melanoma arising in the eustachian tube. AB - Mucosal melanoma is a very rare disease. Most cases have their origins in the head and neck region. To date,only three cases of melanoma originating from the eustachian tube have been reported. We present a case of mucosal melanoma of eustachian tube origin in which a complete excision was performed. In this case, the patient underwent adjuvant radiotherapy and the mass size greatly decreased, which aided subsequent surgical excision. Systemic chemotherapy was not utilized in this case; however, it is often used for palliative purposes. A literature review is also presented. PMID- 16359580 TI - Use of an intra-operative nerve stimulator in identifying the hypoglossal nerve. AB - Previously, the hypoglossal nerve has not undergone intra-operative monitoring during neck operations in which the nerve is at risk. As society becomes increasingly litigious, this may change. This study describes the technique and the microvoltages used in 10 patients for intra-operative stimulation of the hypoglossal nerve with the Magstim nerve stimulator. We confirm that the technique is possible, simple and safe, with minimal disturbance to the patient, anaesthetist and surgeon. PMID- 16359581 TI - The impact of demand factors, quality of care and access to facilities on contraceptive use in Tanzania. AB - The low contraceptive prevalence rate and the existence of unmet demand for family planning services present a challenge for parties involved in family planning research in Tanzania. The observed situation has been explained by the demand-side variables such as socioeconomic characteristics and cultural values that maintain the demand for large families. A small, but growing body of research is examining the effect of supply-side factors such as quality of care of family planning services on the demand for contraceptives. This paper analyses the demand and supply factors determining contraceptive use in Tanzania using the Tanzania Service Availability Survey (1996) and the Tanzania Demographic and Health Survey (1996) data sets. The results show that access to family planning services and quality of care of services are important determinants of contraceptive use in Tanzania even after controlling for demand-side factors. PMID- 16359582 TI - Changes in syntaxin-1B mRNA in the nucleus accumbens of amphetamine-sensitized rats. AB - Repeated administration of amphetamine in animals induces persistent changes in dopamine (DA) functions and behaviour. These changes may be mediated by altered plasticity of the mesocorticolimbic DA system. We have previously reported changes in the content of axonal plasma membrane protein syntaxin-1 in the nucleus accumbens (NAc) of amphetamine-sensitized rats. In the present investigation, we investigated whether syntaxin-1 changes are derived from transcriptional events, i.e. mRNA changes, in the mesocorticolimbic DA regions of the brain. Behavioural sensitization was induced in adult Sprague-Dawley rats by repeated intermittent administration of d-amphetamine (1.5 mg/kg i.p. for 5 alternate days). The animals were sacrificed 24 h, 7 d or 14 d after the last injection and in-situ hybridization using oligonucleotide probes was employed to assess the expression of syntaxin-1A and -1B mRNAs. Results show that the expression of syntaxin-1B mRNA was significantly increased in the NAc shell (NAcS) region in the amphetamine-sensitized rats 14 d after the drug treatment compared to saline pretreated or untreated control animals No significant change in syntaxin-1B mRNA was observed in animals sacrificed 24 h or 7 d after the sensitizing regimen of amphetamine in any brain region analysed, namely NAcS, NAc core (NAcC), caudate putamen (CPu), ventral tegmental area (VTA) or medial prefrontal cortex (mPFC). Levels of syntaxin1A mRNA were not significantly different from controls in any brain region at any time-point. These results suggest that syntaxin-1 protein changes in amphetamine-sensitized rats may be due to increased syntaxin-1B gene expression within local neurons of the NAcS that may lead to altered exocytosis from these neurons during the expression of sensitized response. PMID- 16359583 TI - Discontinuation symptoms in depression and anxiety disorders. AB - The present overview investigates whether different antidepressants have differing discontinuation symptoms upon treatment cessation, if these symptoms vary between depression and anxiety disorders, and with length of treatment. Data came from two comparative studies of escitalopram in major depressive disorder (MDD) (one vs. venlafaxine XR and one vs. paroxetine), two studies in social anxiety disorder (SAD) (one of which used paroxetine as the active reference), and one study in generalized anxiety disorder (GAD), using paroxetine as an active reference [total number of patients: escitalopram (n=1051); paroxetine (n=336); venlafaxine XR (n=124); placebo (n=239)]. All studies included a defined discontinuation period and used the Discontinuation Emergent Signs and Symptoms (DESS) checklist to record the number of discontinuation symptoms. All three antidepressants showed more discontinuation symptoms compared with placebo (p<0.001). Patients reported significantly fewer discontinuation symptoms with escitalopram than with paroxetine and venlafaxine XR in MDD (p<0.05). Escitalopram showed significantly fewer discontinuation symptoms than paroxetine in SAD (p<0.05) and GAD (p<0.001). For each antidepressant, no differences in discontinuation symptoms were observed between the three indications and there was no evidence for increased symptom incidence with increased length of treatment. Thus, discontinuation profiles differ between antidepressants of the same class and are broadly similar in different disorders. No evidence was seen for a higher discontinuation burden with longer treatment. PMID- 16359584 TI - Lymphocyte G-protein receptor kinase (GRK)3 mRNA levels in bipolar disorder. AB - Linkage studies in bipolar disorder were positive for markers in the region of chromosome 22q12.1 including the gene coding for G-protein receptor kinase (GRK)3. Two of six variants of the GRK3 5'-UTR/promoter were reported to be associated with bipolar disorder. GRK3 protein levels in lymphoblastoid cell lines derived from bipolar patients originating from families with linkage to chromosome 22q11 were reported to be decreased compared to those of control subjects and correlated with disease severity. We compared GRK3 mRNA levels in fresh lymphocytes from 31 bipolar patients vs. 26 control subjects, using real time RT-PCR. No overall difference was found between patients and controls. However, GRK3 mRNA levels were markedly and significantly reduced in the subgroup of patients with no family history of a major psychiatric disorder compared with patients with family history. PMID- 16359585 TI - Cerebral palsy: synergism, pathways, and prevention. PMID- 16359587 TI - Infantile postural asymmetry and osteopathic treatment: a randomized therapeutic trial. AB - The aim of this study was to assess the therapeutic efficacy of osteopathic treatment in infants with postural asymmetry. A randomized clinical trial of efficacy with blinded videoscoring was performed. Sixty-one infants with postural asymmetry aged 6 to 12 weeks (mean 9wks) were recruited. Thirty-two infants (18 males, 14 females) with a gestational age of at least 36 weeks were found to be eligible and randomly assigned to the intervention groups, 16 receiving osteopathic treatment and 16 sham therapy. After a treatment period of 4 weeks the outcome was measured using a standardized scale (4-24 points). With sham therapy, five infants improved (at least 3 points), eight infants were unchanged (within 3 points), and three infants deteriorated (not more than -3 points); the mean improvement was 1.2 points (SD 3.5). In the osteopathic group, 13 infants improved and three remained unchanged; the mean improvement was 5.9 points (SD 3.8). The difference was significant (p=0.001). We conclude that osteopathic treatment in the first months of life improves the degree of asymmetry in infants with postural asymmetry. PMID- 16359588 TI - Treatment of adductor spasticity with BTX-A in children with CP: a randomized, double-blind, placebo-controlled study. AB - Adductor spasticity in children with cerebral palsy (CP) impairs motor function and development. In a placebo-controlled, double-blind, randomized multicentre study, we evaluated the effects of botulinum toxin A(BTX-A) in 61 children (37 males, 24 females; mean age 6 years 1 month [SD 3y 1mo]) with CP (leg-dominated tetraparesis, n=39; tetraparesis, n=22; GMFCS level I, n=3; II, n=6; III, n=17; IV, n=29; V, n=6). Four weeks after treatment, a significant superiority of BTX-A was observed in the primary outcome measure (knee-knee distance 'fast catch', p=0.002), the Ashworth scale (p=0.001), and the Goal Attainment Scale (p=0.037). PMID- 16359589 TI - Factor V Leiden mutation: a contributory factor for cerebral palsy? AB - Fifty-seven children with cerebral palsy (CP) and imaging evidence of vascular thrombosis (study group) and 167 children with CP and other imaging finds (control group)were selected. Sixty-one per cent of the study group were male and 53 (93%) had spastic hemiplegia compared with the control group, of whom 55% were male and 54 (32%) had a diagnosis of spastic hemiplegia. Mean age was 5 years 11 months (SD 5y 1mo) for the study group and 7 years 7 months (SD 4y 7mo) for the control group. Blood spots on Guthrie cards or buccal swabs were used to test both groups and their mothers for the factor V Leiden (fVL) mutation, which predisposes carriers to thrombophilia. Mothers were interviewed to gather antenatal, perinatal, demographic, and socio-economic data. The frequency of the fVL mutation in children with evidence of vascular thrombosis and their mothers was not statistically different from the frequency in children with CP with other imaging findings and their mothers. The frequency of the fVL mutation was significantly higher than the expected population frequency of 4% in the study group (10.5%, p=0.012) and in mothers of the control group (7.2%, p=0.036). PMID- 16359590 TI - Validity and reproducibility of a new diagnostic motor performance test in children with suspected myopathy. AB - To spare more children from painful muscle biopsy, a new non-invasive diagnostic motor performance test is undergoing development. Fifteen functional items were used to measure muscle strength and muscle endurance in 68 patients (47 males, 21 females; mean age 7y 8mo, SD 2y 2mo; range 4 to 11y), who had been referred to our specialist centre in the past 3 years on suspicion of myopathy. All the patients had undergone muscle biopsy. To correct the patients' outcomes for age, sex, and body size, regression prediction equations were obtained from a stratified random sample of 64 normally developing primary-school children aged 4 to 11 years (32 males, 32 females; mean age 8y 1mo, SD 2y 4mo). Feasibility was evaluated on the basis of five criteria. Validity was assessed using logistic regression analysis, receiver operating characteristic analysis, and sensitivity and specificity at a specifically chosen cut-off point. Reproducibility was evaluated by test-retest reliability in a stratified random sample of 40 patients who returned for re-measurements using the intraclass correlation coefficient. Seven items satisfied all five feasibility criteria, had high diagnostic power, and high test-retest reliability. The motor performance test can improve diagnostic procedure in children suspected of having myopathy. PMID- 16359591 TI - Naming skills of children born preterm in comparison with their term peers at the ages of 9 and 16 years. AB - The linguistic abilities of children born preterm at 32 weeks' gestation or earlier at Kuopio University Hospital during 1984 to 1986 were evaluated during successive phases of a prospective study. The study protocol included the Rapid Automatic Naming test and Wechsler Intelligence Scale for Children - Revised at 9 years of age and a modified Stroop Color-Word test and the Wechsler Intelligence Scale - Revised at the age of 16 years. Fifty-one children born preterm (26 males, 25 females) and 51 age-matched and sex-matched term controls (26 males, 25 females) were studied at the age of 9 years. At the age of 16 years, 40 children born preterm (19 males, 21 females) and 31 term controls (14 males, 17 females) participated in the study. The children born preterm scored significantly lower in two naming tasks than the controls at the age of 9 years. However, there was no difference between the study groups in naming skills at the age of 16 years or in verbal IQ in either study phase. Maternal education level was not associated with naming skills. Thus, the consequences of preterm birth seem to be minor in relation to linguistic skills during school age and diminish by adolescence. PMID- 16359592 TI - Sensorimotor performance in school-age children with autism, developmental language disorder, or low IQ. AB - The purpose of the study was to determine the prevalence of 'soft' motor deficits in school-aged children with either developmental language disorder (DLD), autism (with high IQ [HiAD] or low IQ [LoAD]), or low IQ without autism (LoIQ), and to evaluate the utility of a refined neurological examination to discriminate between these groups. A total of 242 children (74% male), aged 7 or 9 years, were evaluated as part of a longitudinal, multi-institutional study, with a standardized neurological examination that included Denckla's Physical and Neurological Examination for Soft Signs. Most of the scores separated children into two groups defined by nonverbal IQ, with the DLD and HiAD groups performing better than the LoAD and LoIQ groups. Exceptions included motor impersistence and stereotypies, which were more likely in the autistic groups. The neurologists' summary clinical impressions indicated better sensory/motor skills, oromotor skills, and praxis in the HiAD than in the DLD children. Inability/unwillingness to perform tasks was much more frequent in LoAD than LoIQ children. PMID- 16359593 TI - Autism spectrum disorders and underlying brain pathology in CHARGE association. AB - The rate of autism spectrum disorders (ASDs) and brain abnormalities was analyzed in 31 individuals (15 males, 16 females; age range 1mo to 31y, mean age 8y 11mo) with CHARGE association, as part of a multidisciplinary study. A meticulous neuropsychiatric examination was performed, including standardized autism diagnostic interviews. Judgement regarding ASDs was impossible in three infants and three patients who were deaf and blind. Five individuals met diagnostic criteria for autism, five for an autistic-like condition, and seven for autistic traits. Brain abnormalities were indicated in almost three-quarters of examined individuals, and midline abnormalities of the forebrain in one-third. Awareness of the coexistence of CHARGE and ASDs is important in habilitation care in CHARGE. Moreover, the results indicate that a subgroup of ASDs may be associated with errors in early embryonic brain development. PMID- 16359594 TI - Severe falciparum malaria and acquired childhood language disorder. AB - Language disorders have been reported after severe falciparum malaria but the deficits have not been described in detail. We assessed language outcome in three groups of children aged 6 to 9 years (n=487): those previously admitted to Kilifi District Hospital, Kenya, with cerebral malaria (CM; n=152; mean age 7y 4 mo [SD 1y 1mo]; 77 males, 75 females); or those with malaria and complicated seizures (M/S; n=156; mean age 7y 4mo [SD 1y 2mo]; 72 males, 84 females); and those unexposed to either condition (n=179; mean age 7y 6mo [SD 1y 1mo]; 93 males, 86 females). Median age at hospital admission was 28 months (interquartile range [IQR] 19 to 44 mo) among children with a history of CM and 23 months (IQR 12 to 35mo) among children with a history of M/S. A battery of eight assessments covering the major facets of speech and language was used to measure language performance. Cognitive performance, neurological/motor skills, behaviour, hearing, and vision were also measured. Eighteen (11.8%) of the CM group, 14 (9%) of the M/S group, and four (2.2%) of the unexposed group were found to have a language impairment. CM (odds ratio 3.68, 95% confidence interval 1.09 to 12.4, p=0.04) was associated with significantly increased odds of an impairment-level score relative to the unexposed group. The results suggest that falciparum malaria is one of the most common causes of acquired language disorders in the tropics. PMID- 16359595 TI - Renal vascular disease in neurofibromatosis type 2: association or coincidence? AB - Neurofibromatosis type 2 (NF2) remains a challenging diagnosis in childhood where there may be no neurological involvement. A 12-month-old male in whom NF2 was suspected because of characteristic ophthalmological and cutaneous lesions is reported. Cranial MRI showed no tumours. A pathogenic mutation was identified on NF2 gene analysis. The child developed hypertension due to renal vascular disease. Although renal vascular disease is a recognized complication of neurofibromatosis type 1 (NF1), it has not been reported in NF2. PMID- 16359596 TI - Herpes simplex encephalitis: diagnostic problems and late relapse. AB - A 5-year-old female presented with prolonged afebrile right-sided focal seizures, right brachio-facial paralysis, and dysarthria; consciousness was not altered. Fever appeared 20 hours after onset of neurological symptoms. At admission (day 1) cerebral computerized tomography and cerebrospinal fluid (CSF) analyses were normal including undetectable alpha-interferon (alpha-IFN) and negative herpes simplex virus (HSV) polymerase chain reaction (PCR). Acyclovir was started at a dosage of 60mg/kg/day for 21 days and neurological symptoms improved. Cerebral magnetic resonance imaging (MRI) showed lesions in the left thalamus and left parietal lobe. On day 8, CSF contained an elevated leukocyte count with a predominance of lymphocytes, but alpha-IFN and HSV DNA were still undetectable. Delayed intrathecal synthesis of specific anti-HSV antibodies was found on day 26 and confirmed herpes simplex encephalitis (HSE) diagnosis. Twenty months after this episode, the patient presented with a febrile meningeal syndrome. PCR detected HSV DNA in CSF and cerebral imaging showed a new left temporal lesion. At relapse onset, intrathecal synthesis of specific anti-HSV antibodies had disappeared. Acyclovir was started at a dosage of 60mg/kg/day for 21 days and neurological status improved. At discharge, neurological examination showed right hemiparesis and bucco-facial dyspraxia. Diagnostic problems of HSE diagnosis in children are highlighted. It is suggested that the premature disappearance of intrathecal synthesis of a specific anti-HSV antibody might play a permissive role in the resurgence of cerebral viral replication. PMID- 16359597 TI - Clinical assessment of spasticity in children with cerebral palsy: a critical review of available instruments. AB - This study reviews the instruments used for the clinical assessment of spasticity in children with cerebral palsy, and evaluates their compliance with the concept of spasticity, defined as a velocity-dependent increase in muscle tone to passive stretch. Searches were performed in Medline, Embase, and Cinahl, including the keywords 'spasticity', 'child', and 'cerebral palsy', to identify articles in which a clinical method to measure spasticity was reported. Thirteen clinical spasticity assessment instruments were identified and evaluated using predetermined criteria. This review consists of reports on the standardization applied for assessment at different velocities, testing posture, and quantification of spasticity. Results show that most instruments do not comply with the concept of spasticity; standardization of assessment method is often lacking, and scoring systems of most instruments are ambiguous. Only the Tardieu Scale complies with the concept of spasticity, but this instrument has a comprehensive and time-consuming clinical scoring system. PMID- 16359598 TI - Balancing certainty and uncertainty in clinical medicine. AB - Nothing in clinical medicine is one hundred per cent certain. Part of a doctor's education involves learning how to cope with the anxiety that uncertainty in decisions affecting life and death inevitably produces. This paper examines: (1) the role of anxiety -- both rational and irrational -- in the provision of health care; (2) the effects of uncertainty upon the doctor-patient relationship; (3) the threat uncertainty poses to medical authority (and the assumption of infallibility that props it up); (4) the contribution of clinical uncertainty to the rising popularity of alternative therapies; and (5) the clash between the medical and the legal understanding of how certainty should be defined, particularly as it affects the paediatric community. It concludes by suggesting some strategies that might facilitate successful navigation between the opposing and ever-present forces of certainty and uncertainty. PMID- 16359599 TI - Absence of reference to progressive musculoskeletal pathology in definition of cerebral palsy. PMID- 16359600 TI - Proposed new definition of cerebral palsy does not solve any of the problems of existing definitions. PMID- 16359603 TI - Epilepsy responds to vagus nerve stimulation in ring chromosome 20 syndrome. PMID- 16359604 TI - The correlation between striatal dopamine D2/D3 receptor availability and verbal intelligence quotient in healthy volunteers. AB - BACKGROUND: Although a correlation between the central dopaminergic system and intelligence may exist, the results from imaging studies remain inconclusive. The aim of this study was to explore the relationship between striatal dopamine D2/D3 receptor availability and verbal intelligence quotient (VIQ) using single photon emission computed tomography (SPECT). METHOD: Striatal D2/D3 receptor availability of 64 healthy subjects was determined with the [123I]iodobenzamide ([123I]IBZM) ligand. Intelligence quotients (IQs) of the subjects were measured by the Wechsler Adult Intelligence Scale--Revised (WAIS-R). RESULTS: In addition to age, left striatal D2/D3 receptor availability correlated positively with VIQ. In females, left striatal D2/D3 receptor availability was the only variable that correlated significantly with the similarities subtest of VIQ. CONCLUSIONS: There is a relationship between left striatal D2/D3 receptor availability and verbal intelligence, which varies, predominantly in males. PMID- 16359605 TI - Impaired implicit sequence learning in depression: a probe for frontostriatal dysfunction? AB - BACKGROUND: Implicit learning through motor sequencing tasks is sensitive to basal ganglia dysfunction. Consequently, it is ideally suited for testing elements of the frontostriatal model of major depression and performance can be related to key clinical, neuropsychological, vascular and biochemical data. METHOD: Twenty-one subjects with moderate to severe unipolar depression and 21 age-, sex- and education-matched controls were recruited. Clinical, vascular and biochemical data were recorded. Subjects were administered a battery of neuropsychological tests that assessed speed of processing, working memory, learning, memory, language, perceptual organization and executive functioning. Additionally, subjects were administered a motor sequencing implicit learning task. Implicit learning is assumed when reaction times improve during the sequenced condition as compared to the pseudo-random baseline condition. RESULTS: The rate of implicit learning in persons with depression was only half that of control subjects (3.6% v. 7.3%). Lower rates of implicit learning in patients were associated with poorer performance on neuropsychological tests of visuomotor speed and mental flexibility, longer duration of depressive episode and severity of acute stress. In a small number of subjects, poorer performance was also related to past suicide attempt. CONCLUSIONS: Impaired implicit learning in persons with depression is consistent with frontostriatal dysfunction. Performance is related to some clinical characteristics and to neuropsychological functioning on tests of visuomotor speed and mental flexibility. PMID- 16359608 TI - Bistability and hysteresis in epigenetic regulation of the lactose operon. Since Delbruck, a long series of ignored models. AB - Bistability is the capacity of a system to switch in an "all-or-none" manner between alternative steady states. This powerful concept originates from the analysis of non-linear equations driving open systems. It is one of the various patterns of regulation associated with a particular class of dynamic structures that Glansdorff and Prigogine baptised "dissipative structures". The idea of discontinuous transitions between alternative states was first formulated much earlier, by Delbruck, in 1949. Cohn and Horibata and Novick and Weiner confirmed that such transitions occur in experiments on the lactose operon carried out ten years later. Modelling with non-linear differential equations made it possible to simulate the dynamic behaviour of the lac operon, and modelling by asynchronous logical analysis elucidated the determinant role played by positive feedback circuits in the emergence of multistationarity. Nevertheless, these studies were largely ignored until the recent demonstration of the hysteretic nature of the bistable transition between alternative states of the lac operon. As originally suggested by Delbruck, the pattern of lactose consumption adopted by the bacterium is controlled epigenetically rather than genetically: the true key determinant is the direction of change of an environmental variable with respect to the structural components of the operon. PMID- 16359609 TI - Henrik Kacser--a neglected author? AB - All sciences progress by elaborating new concepts and by finding new relationships between the parts of a system of interacting components. Henrik Kacser, with a background in Biophysics and Genetics, was well placed to develop a new and comprehensive analysis of the control of metabolic fluxes and metabolite concentrations; and to demonstrate that changes in the concentration of normal metabolites were as important in Genetics as changes in the composition of metabolites in bringing about a change in trait. PMID- 16359610 TI - Walter Elsasser, prophet of biological complexity, seeker of simplifying rules. AB - Walter Elsasser was an eminent theoretical physicist who devoted much of his spare time for over half a century (approximately 1935-1987) to the development of a holistic theory of organisms. The three basic principles of his biological theory are a) order in the large which dominates heterogeneity in the small; b) creative selection of the relatively small number of cells and organisms in nature as compared with the immense number of potential molecular states allowed by quantum mechanics, based on the number of atoms, molecules and chemical bonds in a cell, and c) holistic memory in reproduction of cells and organisms, a process which is supplementary to, and fundamentally different in type from the information stored in DNA. His ideas have either been coolly received or ignored by biologists, at least partly because of their purely formal character, although that is considered the sine qua non for theories in physical science. I cite a variety of experiments at the cellular level that illustrate his principles. Those experiments add a concrete dimension to his abstractions that hopefully will promote a dialog between theory and practice to facilitate development of a non-reductionist biology. PMID- 16359611 TI - From semi-conductors to the rhythms of sensitive plants: the research of J.C. Bose. AB - J.C. Bose (1858-1937) was one of the world's first biophysicists. He was the first person to use a semi conducting crystal to detect radio waves, and the ingenious inventor of a portable apparatus for generating and detecting microwaves (approximately 1 cm to 5 mm radio waves, frequency 12-60 GHz), as well as inventing many instruments now routinely used in microwave technology. Bose extended his specialist knowledge of the physics of electromagnetic radiation into insightful experiments on the life-processes of plants. He became a controversial figure in the west. He invented unique, delicate instruments for simultaneously measuring bioelectric potentials and for quantifying very small movements in plants. He worked with touch-sensitive plants, including Mimosa pudica, with plants that perform spontaneous movements, including the Indian telegraph plant Desmodium, and with plants and trees that did not make obvious rapid movements. Bose concluded that plants and animals have essentially the same fundamental physiological mechanisms. All plants co-ordinate their movements and responses to the environment through electrical signalling. All plants are sensitive explorers of their world, responding to it through a fundamental, pulsatile, motif involving coupled oscillations in electric potential, turgor pressure, contractility, and growth. His overall conclusion that plants have an electromechanical pulse, a nervous system, a form of intelligence, and are capable of remembering and learning, was not well received in its time. A hundred years later, concepts of plant intelligence, learning, and long-distance electrical signalling in plants have entered the mainstream literature. PMID- 16359612 TI - How membrane carriers function: the critical contribution of W.F. Widdas (1952) to modern understanding. AB - This paper reviews the contribution to the field of transport biology of W.F. Widdas' paper 'Inability of diffusion to account for placental glucose transfer in the sheep and consideration of the kinetics of a possible carrier transfer' published in the Journal of Physiology in 1952. The importance of this mathematical analysis, and of its assumptions and predictions, has been supported in a number of subsequent publications over the last half century. But its vindication emerges specifically from the recent successful solution of the three dimensional structure of a membrane transport protein, the sugar carrier lac permease. Predictions arising from the 1952 paper are confirmed by this very recent study. Reasons for the relatively slow acceptance of the Widdas carrier hypothesis and its implications are discussed. PMID- 16359613 TI - Henri Atlan's early writings. AB - The importance given to biological complexity and dynamics, the progressive giving up the notion of genetic programme, the attention paid to noise at the molecular level, are all new aspects of biological research characteristic of the post-genomic era. They are frequently mentioned as the signs of the delayed recognition of the important contributions made by Henri Atlan from the beginning of the 1970s. By focusing my study on his first major essay, l'Organisation Biologique et la Theorie de l'Information, I reach a somewhat different conclusion. Whereas Henri Atlan proposed an early criticism of the notion of genetic programme, the point of view he adopted was at odds with molecular biology. His attempt to apply the theory of information to organisms was a project already abandoned in the United States. The importance he placed to the second law of thermodynamics and his insistence on the limits of the Darwinian theory of evolution were more the sign of an attachment to past traditions of biological thought, still permeating the French community of biologists in the second half of the XXth century, than an anticipation of future developments in the post-genomic era. PMID- 16359614 TI - Poly-(R)-3-hydroxybutyrate and the pioneering work of Rosetta Natoli Reusch. AB - In investigating genetic competence, Reusch and collaborators have found that the concentration of short chain poly-(R)-3-hydroxybutyrate (PHB) and polyphosphate (polyP) complexes increases with genetic transformability and that interrupting DNA uptake yields single-stranded donor DNA complexed with short chain PHB. This would be consistent with the organic polyphosphate, DNA, replacing the inorganic polyphosphate, polyP, in the PHB pore so allowing the DNA to be drawn into the cell. Reusch has gone on to show that PHB and polyphosphate, extracted from membranes or synthesized chemically, together form a voltage-activated calcium selective channel. One may wonder whether the classical proteinaceous calcium channels have a short chain PHB/polyP core--and whether other ion channels have this core too. It is therefore significant that in Streptomyces lividans the potassium channel KcsA, which resembles that of eukaryotes, forms tetramers that contain polyP whilst both monomers and tetramers are covalently linked to short chain PHB. Pumps are the counterparts of channels. Reusch has also shown that a model pump, the calcium ATPase pump of human erythrocytes, contains both cPHB and polyP and has strongly implicated these polymers in its functioning. Again, one may wonder whether these polymers are essential constituents of other pumps. Reusch has gone on to show that a wide range of proteins are modified post translationally by covalent addition of short chain PHB in both prokaryotes and eukaryotes including DNA-binding proteins such as histones. Finally, Reusch has extended the importance of short chain PHB to medicine by showing its likely involvement in atherogenic plaques and diabetes. And yet this opus has gone largely unnoticed. PMID- 16359615 TI - Serendipity strikes twice: the discovery and rediscovery of defective ribosomal products (DRiPS). AB - Here I recount how the remarkably large fraction of rapidly degraded nascent proteins was discovered through serendipity not once, but twice. This story is used as a canvas for discussing how the imperfections in human nature retard scientific progress. PMID- 16359616 TI - Historical blunders: how toxicology got the dose-response relationship half right. AB - Substantial evidence indicates that reliable examples of hormetic dose responses in the toxicological literature are common and generalizable across biological model, endpoint measured and chemical class. Further evaluation revealed that the hormetic dose response model is more common than the threshold dose response model in objective, head-to-head comparisons. Nonetheless, the field of toxicology made a profound error by rejecting the use of the hormetic dose response model in its teaching, research, risk assessment and regulatory activities over nearly the past century. This paper argues that the hormetic dose response model (formerly called the Arndt-Schulz Law) was rejected principally because of its close historical association with the medical practice of homeopathy as a result of the prolonged and bitter feud between traditional medicine and homeopathy. Opponents of the concept of hormesis, making use of strong appeals to authority, were successful in their misrepresentation of the scientific foundations of hormesis and in their unfair association of it with segments of the homeopathic movement with extreme and discreditable views. These misrepresentations became established and integrated within the pharmacology and toxicology communities as a result of their origins in and continuities with traditional medicine and subsequently profoundly impacted a broad range of governmental risk assessment activities further consolidating the rejection of hormesis. This error of judgment was reinforced by toxicological hazard assessment methods using only high and few doses that were unable to assess hormetic responses, statistical modeling processes that were constrained to deny the possibility of hormetic dose response relationships and by the modest nature of the hormetic stimulatory response itself, which required more rigorous study designs to evaluate possible hormetic responses. PMID- 16359617 TI - Is biomedical nuclear magnetic resonance limited by a revisitable paradigm in physics? AB - The history of nuclear magnetic resonance (NMR) can be divided generally into two phases: before the Second World War, molecular beam methods made it possible to detect the whole set of spins. However, these methods were destructive for the sample and had a very low precision. The publications of F. Bloch and E. Purcell in 1946 opened up a second phase for NMR with the study of condensed matter, but at the expense of an enormous loss in theoretical sensitivity. During more than half a century, the method of Bloch and Purcell, based on inductive detection of the NMR signal, has allowed many developments in biomedicine. But, curiously, this severely constraining limitation on sensitivity has not been called into question during this half-century, as if the pioneers of the pre-war period had been forgotten. PMID- 16359618 TI - Biological oxidation: over a century of hardship for the concept of active oxygen. AB - After much research in the field of biological oxidation, two main concepts of the mechanism of energy gain were consecutively proposed--the concept of direct oxygen activation (analogous to combustion), and the concept of activation of hydrogen followed by gradual energy release (analogous to putrefaction). The latter concept currently dominates. According to it, practically all the energy required in life is gained initially in the form of ATP; the latter being synthesized mostly due to the classical oxidative phosphorylation process. Direct activation of oxygen is considered to be a secondary pathway in its utilization, unrelated to bioenergetics. Besides, this pathway seems to be risky, since reactive oxygen species (ROS) are produced on account of it. However, evidence has been accumulated that a considerable part of all oxygen consumption undergoes direct activation; and this share increases with energy demand. Recent data also argues that ROS are indispensable for regulating biological function at all levels of biological organization. In spite of intense activation of oxygen, the usual levels of ROS in cells and tissues are extremely low due to their immediate elimination by the powerful an "antioxidant defense" system. In the process of elimination of ROS, high density energy (energy of electronic activation), equivalent to the energy of photons of visible and UV-light, is necessarily released, which has a much higher grade than the energy released in putrefaction. Both previous and more recent data argue that the energy of electronic excitation is biologically significant. The role of an aqueous environment with unusual water structuring in managing this form of energy in biology will also be discussed. Thus, the processes of oxygen activation in aqueous systems seem to be no less fundamental than the processes of activation of hydrogen atoms in organic fuels. Current bioenergetics has to consider both oxygen and hydrogen activation rather than concentrate on only one mode of energy gain and utilization. PMID- 16359619 TI - The state of water in living systems: from the liquid to the jellyfish. AB - The status of water in living systems is reviewed both from philosophical and scientific viewpoints. Starting from antique Mediterranean civilizations (Sumerian, Egyptian, Hebrew, Greek), a world trip is proposed through Norse myths, Siberian Shamanism, Hinduism, Taoism, Buddhism, Shinto, Mayan, Aztec, Inca, Aboriginal and African philosophies in order to convince that all humans share the same qualitative idea that water was a pre-requisite for life apparition. The quantitative aspect of the problem is further analyzed at the light of the scientific contributions from two leading scientists: R.A. Gortner and E.T. Jaynes. With Gortner's work it is demonstrated using the concrete example of the Jellyfish submitted at a Faraday discussion held in London in 1930, how a paradigm shift has occurred in the thirties concerning the status of bound water in the living cell. With Jaynes' work, the disastrous consequences of the entrenchment of diffusion theory in biology are critically examined and the exact meaning of the second law of thermodynamics for biological systems is given using the concrete example of muscle contraction. In conclusion, the importance of distinguishing between an ontological and epistemological level of knowledge is stressed and suggestions for reconciling scientific and philosophical approaches are given. PMID- 16359620 TI - Consciousness, information, and living systems. AB - The possibility of a proactive role for consciousness in the establishment of physical reality has been addressed via an extensive 26-year program investigating physical anomalies in human/machine interactions and non-sensory acquisition of information about remote geographical locations. Empirical databases comprising many hundreds of millions of random events confirm that information can be introduced into, or extracted from, otherwise random physical processes solely through the agencies of human intention and subjective resonance. Much of the evidence mitigates the likelihood that the anomalies are manifestations of neo-cortical cognitive activity. Rather, they may be expressions of a deeper information organizing capacity of biological origin that emerges from the uncertainty inherent in the complexity of all living systems. PMID- 16359621 TI - Protoreaction of protoplasm. AB - My goal is to describe briefly the universal cellular reaction (UCR) to external actions and agents. This general reaction was the main subject of investigation by the scientific school of the outstanding Russian cytologist, Dmitrii Nasonov (1895-1957). The UCR consists of two phases of complex changes in cellular viscosity and turbidity, in the cell's ability to bind vital dyes, in the resting membrane potential, and in cellular resistance to harmful actions. Works from the Nasonov School have shown that these changes are based on structural-functional transformations of many cell proteins that react uniformly to actions of different physical and chemical nature. In general, these complex changes do not depend on cell type, indicating the universal and ancient nature of the UCR as well as its general biological significance. A new interpretation of the mechanism of the universal reaction is proposed in this paper, and a possible role for contractile proteins in the mechanism of the UCR of muscle cells is presented. In addition, the concept of cell hydrophobicity is introduced. Nasonov's School proposed a concept of physiological standardization that allows comparison of data obtained by different investigators and that will also be described here. PMID- 16359622 TI - Doubts about the sodium-potassium pump are not permissible in modern bioscience. AB - The sodium-potassium pump--the central molecular model of the generally accepted membrane pump theory (MPT)--is a misconception, according to the alternative association-induction hypothesis (AIH) of Gilbert Ling. With a discreet fraud coupled with the practice of appraising scientific publications and grant proposals by the peer-review system, the academic establishment--inadvertently or not--hinders a general discussion and acceptance of decisive arguments of the AIH. As a result, important discoveries over many decades, as well as new ways of basic research promising success remain largely unknown. The habit of funding safe research on ideas that have a false premise persists, and the wrong theory gets ever deeper entrenched. Science suffers. PMID- 16359623 TI - The cohesion of water in biology: a property not to be forgotten. AB - Gases and crystalline solids are states of matter that have been understood for nearly two centuries but liquid as a state of matter is still unclear. As a third state of matter there have been many anomalies uncovered in the twentieth century such as dipoles and different properties from changed molecular structures. In consequence liquids can no longer be grouped into a separate state of matter. Liquid water, the most abundant material in the Universe has a number of discrete characteristics. The liquid surface and the cohesive forces of liquid water are two of the more general properties that have unique importance in molecular biology. It is shown here that Coulomb forces over short distances can stabilize water molecules that are rapidly spinning dipoles or dipoles which have lost their rotational energy and form semi-crystal-like solids when confined to the restricted spaces of cells and proteins. The surface energy of liquids, extensively studied since the 19th century, can do mechanical work and this is clearest with liquid mercury. But it is surprising that this remarkable property has been neglected in the case of water in biology, and particularly not envisaged as a work supplementary to ATP hydrolysis in muscle contractions, which merits further study. PMID- 16359624 TI - The surface energy of water; molecular biology of the working mechanisms. AB - The surface tension and surface energy of water, have been studied for two centuries. The ability to do physical chemical work dates from the decades at the close of the nineteenth century and opening of the twentieth. Teaching and research popularity dropped in the post-war years, and today is practically unique. Before 1846, Laplace, the French scientist, found that pressure was needed to force water, or other liquid, through small holes. The pressure needed was greater for smaller holes, and the relationship depended upon the surface tension of the liquid under test, and the inverse of the radius of the holes. The Laplace formula still appears in Physiological textbooks. The importance in the small air tubes of respiratory physiology, and in the capillary circulation generally, is still studied. The use of surface energy for protein conformational changes has received little attention. A proposed scheme for the human erythrocyte glucose transporter, although shown to be experimentally consistent, has been disregarded. A possible reason is the difficulty of comprehending how surface energy can actually perform mechanical work. Therefore, studies with molecular models, which increase the understanding of how mechanical work is done are described in more detail here. PMID- 16359625 TI - The simultaneous collapse of both the swinging crossbridge theory of muscle contraction and the in vitro motility essays. AB - In the early seventies we discovered that isolated, active, myosin fragments can induce movement and tension generation by actin filaments in both in vitro and in vivo systems, employing a variety of techniques. It was not in line with the domineering swinging crossbridge theory of muscle contraction. We then proposed an hydrodynamic mechanism which explained our results and was applied to muscle contraction and to other biological engines. Our discovery has been ignored for a long time until the so-called "in vitro motility essays" appeared. By using this artifact--laden technique the mechanochemical reactivity of the active myosin fragments was re-discovered without giving us any credit. The essays gave continuously changing values for fundamental parameters of muscle contraction; the values were appreciably different in different laboratories and decreased in a continuous fashion in the hands of one scientist. By analyzing recent experiments which derived the rate of ATP hydrolysis of active muscles as function of the applied load I calculated the value of the sliding distance resulting from the breakdown of one ATP molecule by each of the myosin heads in contracting muscle. According to the contemporary theory this should be the same for all muscles under any environmental conditions and determined by length of the myosin head's neck. My examination led to the conclusion that the sliding distance varies from one muscle to another and with different temperatures for the same muscle. This again, contradicts the current theory and should give the final blow to both this theory and the "essays". Furthermore: it can be explained by a hydrodynamic mechanism such as that proposed by us more than 30 years ago. PMID- 16359626 TI - Circulating DNA in higher organisms cancer detection brings back to life an ignored phenomenon. AB - The concept of circulating DNA is derived from the early transformation experiments on bacteria. We describe first experiments done with graft hybrids which could be due to circulating DNA. Work on uptake of foreign DNA by eukaryotic cells is then reported. This work led us to discover the phenomenon of transcession in plants and animals where we showed that bacterial DNA can spontaneously enter cells of eukaryotes and be transcribed. We then outline the fact that living cells spontaneously release DNA within a homeostatic mechanism. Finally we describe in more detail some experiments suggesting that an exchange of DNA between T and B lymphocytes may have immunological implications. In this work, nude mice injected with DNA excreted by antigen stimulated human T lymphocytes produced specific antibodies expressing human characteristics. PMID- 16359627 TI - Concepts, facts and artifacts in electron microscopy. AB - This communication illustrates how the electron microscope has contributed to biochemistry by revealing how multienzyme systems in mitochondria are structurally organized to secure high speed ATP synthesis and has extended physiology to the molecular level. Ribonucleoprotein complexes form a gel in the cytoplasm determining the conditions for translation... Photoreceptor stimulation involves two phases, trapping of light by a light reflecting cylinder formed by the outer segment disks and energy transduction by bleaching of photopigment molecules changing the charge of the outer segment disks driving the photoreceptor toward hyperpolarization. Revealing the synaptic connections between retinal neurons extends neurophysiology to the level of information processing by neural circuits, which are designed for high speed processing. Spatial brightness contrast enhancement is eliminated in connection with macular degeneration, which leads to partial blindness, revealing the importance of contrast enhancement for vision. PMID- 16359628 TI - Assembling the building blocks of cell theory. AB - Changes of scientific paradigms happen with the emergence of new theories, like the cell theory in the 19th century. These new conceptions are the result of a collective work occurring over decades or centuries. Scientists who participate in these important mutations are fully in tune with their time, even when they are considered precursors, and it is always difficult to judge how far they were right and how far they were wrong. PMID- 16359629 TI - "Fathers" and "sons" of theories in cell physiology: the membrane theory. AB - The last 50 years in the history of life sciences are remarkable for a new important feature that looks as a great threat for their future. A profound specialization dominating in quickly developing fields of science causes a crisis of the scientific method. The essence of the method is a unity of two elements, the experimental data and the theory that explains them. To us, "fathers" of science, classically, were the creators of new ideas and theories. They were the true experts of their own theories. It is only they who have the right to say: "I am the theory". In other words, they were carriers of theories, of the theoretical knowledge. The fathers provided the necessary logical integrity to their theories, since theories in biology have still to be based on strict mathematical proofs. It is not true for sons. As a result of massive specialization, modern experts operate in very confined close spaces. They formulate particular rules far from the level of theory. The main theories of science are known to them only at the textbook level. Nowadays, nobody can say: "I am the theory". With whom, then is it possible to discuss today on a broader theoretical level? How can a classical theory--for example, the membrane one--be changed or even disproved under these conditions? How can the "sons" with their narrow education catch sight of membrane theory defects? As a result, "global" theories have few critics and control. Due to specialization, we have lost the ability to work at the experimental level of biology within the correct or appropriate theoretical context. The scientific method in its classic form is now being rapidly eroded. A good case can be made for "Membrane Theory", to which we will largely refer throughout this article. PMID- 16359630 TI - Building bridges between the physical and biological sciences. AB - This paper attempts to identify major conceptual issues that have inhibited the application of physical chemistry to problems in the biological sciences. We will trace out where theories went wrong, how to repair the present foundations, and discuss current progress toward building a better dialogue. PMID- 16359631 TI - Revitalizing science in a risk-averse culture: reflections on the syndrome and prescriptions for its cure. AB - This paper considers problems with the scientific culture and granting systems, the most important of which is an aversion to risk. Grant awards tend to be "safe" rather than bold. This discourages the fresh approaches that may bring important breakthroughs. The paper then suggests remedies that could restore the scientific enterprise to one that is friendlier to fresh thinking. PMID- 16359632 TI - Partial perceptual equivalence between vision and touch for texture information. AB - The present study examined the extent to which vision and touch are perceptually equivalent for texture information in adults. Using Garbin's method, we selected two sets of textures having high versus low cross-modal dissimilarity values between vision and touch (Experiment 1). The two sets of textures were then used as material in a cross-modal matching task (Experiment 2). Results showed that asymmetries occurred in the performances when the stimuli had high cross-modal dissimilarity values, but not when the stimuli had low cross-modal dissimilarity values. These results extend Garbin's findings on shape information to the texture domain and support the idea that partial perceptual equivalence exists between vision and touch. PMID- 16359633 TI - Determination of compound aminopyrine phenacetin tablets by using artificial neural networks combined with principal components analysis. AB - A method for simultaneous, nondestructive analysis of aminopyrine and phenacetin in compound aminopyrine phenacetin tablets with different concentrations has been developed by principal component artificial neural networks (PC-ANNs) on near infrared (NIR) spectroscopy. In PC-ANN models, the spectral data were initially analyzed by principal component analysis. Then the scores of the principal components were chosen as input nodes for the input layer instead of the spectral data. The artificial neural network models using the spectral data as input nodes were also established and compared with the PC-ANN models. Four different preprocessing methods (first-derivative, second-derivative, standard normal variate (SNV), and multiplicative scatter correction) were applied to three sets of NIR spectra of compound aminopyrine phenacetin tablets. The PC-ANNs approach with SNV preprocessing spectra was found to provide the best results. The degree of approximation was performed as the selective criterion of the optimum network parameters. PMID- 16359634 TI - Alu-linked hairpins efficiently mediate RNA interference with less toxicity than do H1-expressed short hairpin RNAs. AB - RNA interference has become a powerful tool for specific inhibition of gene expression in mammalian cells. Expression constructs allow for the long-term delivery of short interfering RNAs, usually through the expression of Pol III transcribed hairpins. In some instances, these expression systems have been shown to have side effects, including induction of the interferon response and cytotoxicity. Here we demonstrate that H1-expressed hairpins, as well as the cloning vector, reduce the plating efficiency of HeLa cells. This toxicity is abrogated by coexpression of the hairpin in the same transcript as a human Alu repetitive element. These Alu-linked hairpins retain the ability to knock down expression of target mRNAs. This modification, which we term SINE (short interspersed repetitive element)-enhanced short hairpin RNA, provides an alternative expression system for hairpins with reduced side effects. PMID- 16359635 TI - Computer simulation for ergonomic improvements in laparoscopic surgery. AB - It is the aim of this study to reduce the stress and strain of the medical staff during laparoscopic operations, and, simultaneously, to increase the safety and efficiency of an integrated operation room (OR) by an ergonomic redesign. This was attempted by a computer simulation approach using free modelling of the OR and 3D human models (manikins). After defining ergonomically "ideal" postures, optimal solutions for key elements of an ergonomic design of the OR (position and height of the image displays, height of the OR table and the Mayo stand) could be evaluated with special regard to the different individual body size of each member of the team. These data should be useful for the development of team adapted, user friendly integrated OR suites of the future. PMID- 16359636 TI - Characterization of pyrethroid hydrolysis by the human liver carboxylesterases hCE-1 and hCE-2. AB - Carboxylesterases hydrolyze a large array of endogenous and exogenous ester containing compounds, including pyrethroid insecticides. Herein, we report the specific activities and kinetic parameters of human carboxylesterase (hCE)-1 and hCE-2 using authentic pyrethroids and pyrethroid-like, fluorescent surrogates. Both hCE-1 and hCE-2 hydrolyzed type I and II pyrethroids with strong stereoselectivity. For example, the trans-isomers of permethrin and cypermethrin were hydrolyzed much faster than corresponding cis-counterparts by both enzymes. Kinetic values of hCE-1 and hCE-2 were determined using cypermethrin and 11 stereoisomers of the pyrethroid-like, fluorescent surrogates. K(m) values for the authentic pyrethroids and fluorescent surrogates were in general lower than those for other ester-containing substrates of hCEs. The pyrethroid-like, fluorescent surrogates were hydrolyzed at rates similar to the authentic pyrethroids by both enzymes, suggesting the potential of these compounds as tools for high throughput screening of esterases that hydrolyze pyrethroids. PMID- 16359637 TI - Differential effects of hyperosmotic challenge on interleukin-1-activated pathways in bovine articular cartilage. AB - Chondrocytes in situ experience fluctuations in extracellular osmolarity resulting from mechanical loading. The objective of this study was to determine whether hyperosmotic stress causes or exacerbates interleukin-1 (IL-1)-mediated effects in bovine articular cartilage. Disks of cartilage cut from the articular surface of calf radiocarpal joints were incubated for 24h in the presence or absence of IL-1 in Dulbecco's modified Eagle's medium adjusted to various osmolalities with sucrose or NaCl. Cyclooxygenase (COX)-2 levels in the cartilage were examined by Western blot. Culture media were assayed for prostaglandin E(2) (PGE(2)), nitrite as an indicator of nitric oxide (NO) production, and sulfated glycosaminoglycan as an indicator of proteoglycan degradation. We report the osmolality-dependent potentiation of COX-2 and PGE(2) production, and the osmolality-dependent inhibition of NO production and proteoglycan degradation in IL-1-activated cartilage. The data demonstrate that osmotic and cytokine signaling interact to differentially modulate IL-1-stimulated effects in calf articular cartilage. PMID- 16359638 TI - Isolation and characterization of a sodium-dependent phosphate transporter gene in Dunaliella viridis. AB - A sodium-dependent phosphate transporter gene, DvSPT1, was isolated from a cDNA library using a probe derived from a subtracted cDNA library of Dunaliella viridis. Sequencing analyses revealed a cDNA sequence of 2649 bp long and encoded an open-reading frame consisting of 672 amino acids. The deduced amino acid sequence of DvSPT1 exhibited 31.2% identity to that of TcPHO from Tetraselmis chui. Hydrophobicity and secondary structure prediction revealed 11 conserved transmembrane domains similar to those found in PHO89 from Saccharomyces cerevisiae and PHO4 from Neurospora crassa. Northern blot analysis indicated that the DvSPT1 expression was induced upon NaCl hyperosmotic stress or phosphate depletion. Functional characterization in yeast Na+ export pump mutant G19 suggested that DvSPT1 encoded a Na+ transporter protein. The gene sequence of GDvSPT1 (7922 bp) was isolated from a genomic library of D. viridis. Southern blot analysis indicated that there exist at least two homologous genes in D. viridis. PMID- 16359639 TI - Methylation changes of H19 gene in sperms of X-irradiated mouse and maintenance in offspring. AB - The nature of imprinting is just differential methylation of imprinted genes. Unlike the non-imprinted genes, the methylation pattern of imprinted genes established during the period of gametogenesis remains unchangeable after fertilization and during embryo development. It implies that gametogenesis is the key stage for methylation pattern of imprinted genes. The imprinting interfered by exogenous factors during this stage could be inherited to offspring and cause genetic effect. Now many studies have proved that ionizing irradiation could disturb DNA methylation. Here we choose BALB/c mice as a research model and X-ray as interfering source to further clarify it. We discovered that the whole-body irradiation of X-ray to male BALB/c mice could influence the methylation pattern of H19 gene in sperms, which resulted in some cytosines of partial CpG islands in the imprinting control region could not transform to methylated cytosines. Furthermore, by copulating the interfered male mice with normal female, we analyzed the promoter methylation pattern of H19 in offspring fetal liver and compared the same to the pattern of male parent in sperms. We found that the majority of methylation changes in offspring liver were related to the ones in their parent sperms. Our data proved that the changes of the H19 gene methylation pattern interfered by X-ray irradiation could be transmitted and maintained in the first-generation offspring. PMID- 16359640 TI - The forgotten Gram-negative bacilli: what genetic determinants are telling us about the spread of antibiotic resistance. AB - Gram-negative bacilli have become increasingly resistant to antibiotics over the past 2 decades due to selective pressure from the extensive use of antibiotics in the hospital and community. In addition, these bacteria have made optimum use of their innate genetic capabilities to extensively mutate structural and regulatory genes of antibiotic resistance factors, broadening their ability to modify or otherwise inactivate antibiotics in the cell. The great genetic plasticity of bacteria have permitted the transfer of resistance genes on plasmids and integrons between bacterial species allowing an unprecedented dissemination of genes leading to broad-spectrum resistance. As a result, many Gram-negative bacilli possess a complicated set of genes encoding efflux pumps, alterations in outer membrane lipopolysaccharides, regulation of porins and drug inactivating enzymes such as beta-lactamases, that diminish the clinical utility of today's antibiotics. The cross-species mobility of these resistance genes indicates that multidrug resistance will only increase in the future, impacting the efficacy of existing antimicrobials. This trend toward greater resistance comes at a time when very few new antibiotics have been identified capable of controlling such multi-antibiotic resistant pathogens. The continued dissemination of these resistance genes underscores the need for new classes of antibiotics that do not possess the liability of cross-resistance to existing classes of drugs and thereby having diminished potency against Gram-negative bacilli. PMID- 16359642 TI - Dihydrofolate reductase inhibitors as antibacterial agents. AB - Although only a few DHFR inhibitors have progressed as antibiotics to the market there is much renewed interest in the discovery and development of new generation DHFR inhibitors as antibacterial agents. This article describes the success in exploiting DHFR as a drugable target as exemplified by trimethoprim (TMP) and the development of several new diaminopyrimidines. Iclaprim, a recent example of a novel diaminopyrimidine currently in Phase III clinical trials, is also described together with several examples of anti-DHFR antibacterial compounds in pre clinical development. PMID- 16359641 TI - Structure activity and molecular modeling analyses of ribose- and base-modified uridine 5'-triphosphate analogues at the human P2Y2 and P2Y4 receptors. AB - With the long-term goal of developing receptor subtype-selective high affinity agonists for the uracil nucleotide-activated P2Y receptors we have carried out a series of structure activity and molecular modeling studies of the human P2Y2 and P2Y4 receptors. UTP analogues with substitutions in the 2'-position of the ribose moiety retained capacity to activate both P2Y2 and P2Y4 receptors. Certain of these analogues were equieffective for activation of both receptors whereas 2' amino-2'-deoxy-UTP exhibited higher potency for the P2Y2 receptor and 2'-azido UTP exhibited higher potency for the P2Y4 receptor. 4-Thio substitution of the uracil base resulted in a UTP analogue with increased potency relative to UTP for activation of both the P2Y2 and P2Y4 receptors. In contrast, 2-thio substitution and halo- or alkyl substitution in the 5-position of the uracil base resulted in molecules that were 3-30-fold more potent at the P2Y2 receptor than P2Y4 receptor. 6-Aza-UTP was a P2Y2 receptor agonist that exhibited no activity at the P2Y4 receptor. Stereoisomers of UTPalphaS and 2'-deoxy-UTPalphaS were more potent at the P2Y2 than P2Y4 receptor, and the R-configuration was favored at both receptors. Molecular docking studies revealed that the binding mode of UTP is similar for both the P2Y2 and P2Y4 receptor binding pockets with the most prominent dissimilarities of the two receptors located in the second transmembrane domain (V90 in the P2Y2 receptor and I92 in the P2Y4 receptor) and the second extracellular loop (T182 in the P2Y2 receptor and L184 in the P2Y4 receptor). In summary, this work reveals substitutions in UTP that differentially affect agonist activity at P2Y2 versus P2Y4 receptors and in combination with molecular modeling studies should lead to chemical synthesis of new receptor subtype-selective drugs. PMID- 16359643 TI - Understanding the longevity of the beta-lactam antibiotics and of antibiotic/beta lactamase inhibitor combinations. AB - Microbial resistance necessitates the search for new targets and new antibiotics. However, it is likely that resistance problems will eventually threaten these new products and it may, therefore, be instructive to review the successful employment of beta-lactam antibiotic/beta-lactamase inhibitor combinations to combat penicillin resistance. These combination drugs have proven successful for more than two decades, with inhibitor resistance still being relatively rare. The beta-lactamase inhibitors are mechanism-based irreversible inactivators. The ability of the inhibitors to avoid resistance may be due to the structural similarities between the substrate and inhibitor. PMID- 16359644 TI - Effects of ATP and GTP on voltage-gated K+ currents in glandular and muscular sympathetic neurons. AB - This study assesses the effects of ATP and GTP on the kinetic properties of voltage-gated K+ currents in anatomically identified postganglionic sympathetic neurons innervating the submandibular gland and the masseter muscle in rats. Three types of K+ currents were isolated: the I(Af) steady-state inactivating at more hyperpolarized potentials, I(As) steady-state inactivating at less hyperpolarized potentials than I(Af) and the I(K) current independent of membrane potential. The kinetic properties of these currents were tested in neurons with ATP (4 mM) and GTP (0.5 mM) or without ATP and GTP in the intracellular solution. In glandular and muscular neurons in the absence of ATP and GTP in the intracellular solution, the current density of I(Af) was significantly larger (142 pA/pF and 166 pA/pF, respectively) comparing to cells with ATP and GTP (96 pA/pF and 100 pA/pF, respectively). The I(As) was larger only in glandular neurons (52 pA/pF vs. 37 pA/pF).Conversely, I(K) current density was smaller in glandular and muscular neurons without ATP and GTP (17 pA/pF and 31 pA/pF, respectively) comparing to cells with ATP and GTP (57 pA/pF and 58 pA/pF, respectively). In glandular (15.5 nA/ms vs. 6.9 nA/ms) and muscular (10.9 nA/ms vs. 7.5 nA/ms) neurons, the I(Af) activated faster in the absence of ATP and GTP. Half inactivation voltage of I(Af) in glandular (-110.0 mV vs. -119.7 mV) and muscular (-108.4 vs. -117.3 mV) neurons was shifted towards depolarization in the absence of ATP and GTP. We suggest that the kinetic properties of K+ currents in glandular and muscular sympathetic neurons change markedly in the absence of ATP and GTP in the cytoplasm. Effectiveness of steady-state inactivated currents (I(Af) and I(AS)) increased, while effectiveness of steady-state noninactivated currents decreased in the absence of ATP and GTP. The effects were more pronounced in glandular than in muscular neurons. PMID- 16359645 TI - Developmental expression of neuronal nitric oxide synthase in P/Q-type voltage gated calcium ion channel mutant mice, leaner and tottering. AB - Nitric oxide (NO) is a diffusible messenger molecule produced primarily by neuronal nitric oxide synthase (nNOS) in the central nervous system. Both nNOS expression and NO production are regulated by calcium ions. Leaner and tottering mice carry a mutation in the pore forming subunit (alpha1A) of P/Q-type voltage gated calcium ion channels, which decreases calcium ion current through the affected channels and disrupts calcium homeostasis. We have previously shown that nNOS expression is altered in adult leaner and tottering cerebella. In addition, leaner and tottering mice have been shown to have abnormal cerebellar granule cell-Purkinje cell synapses and leaner cerebellar granule cells undergo abnormal apoptosis during early postnatal development. Since NO production has been linked to several developmental roles including neuronal cell death, synaptogenesis and neuronal cell survival, our objective was to evaluate the expression of nNOS in developing leaner and tottering cerebella. Our results show that nNOS is differentially expressed in leaner and tottering cerebella compared to wild type cerebella and compared to each other. In whole cerebella, Western blotting revealed a significant increase in nNOS expression at postnatal day 12 in tottering but not leaner or wild type cerebella. At the cellular level the NADPH diaphorase marker for nNOS revealed a significant increase in nNOS expression in basket cell interneurons in both mutant mice. nNOS expression in granule cells in the internal granule cell layer in tottering mice was increased at P12, while granule cells of leaner mice exhibited decreased nNOS expression at P20. The changes in nNOS expression at P12 did not correlate with a change in overall NO production, but rather maintained wild type NO concentrations. These findings suggest that changes in nNOS expression in the leaner and tottering cerebella are compensatory in nature with NO most likely functioning as a calcium-regulated neuroprotective/neurotrophic factor in postnatal cerebellar development. PMID- 16359646 TI - Ethanol prevents NMDA receptor reduction by maternal separation in neonatal rat hippocampus. AB - We measured the effects of ethanol on glutamate receptor levels in the hippocampus of neonatal Wistar rats using a vapor chamber model. Two control groups were used; a normal suckle group and a maternal separation group. Levels of NMDA receptors were not significantly altered in ethanol-treated animals compared to the normal suckle control group, as shown by [3H]MK-801 binding and Western blot analysis. However, MK-801 binding and NR1 subunit immunoreactivity were greatly reduced in the hippocampus of separation control animals. Neither ethanol treatment nor maternal separation altered levels of GluR1 or GluR2(4). These results have serious implications for the importance of maternal contact for normal brain development. PMID- 16359647 TI - When artificial grammar acquisition in Parkinson's disease is impaired: the case of learning via trial-by-trial feedback. AB - Although there is strong evidence that human category learning is mediated by qualitatively distinct systems, the neural substrates of procedural category learning remain largely unclear. The present research sought to investigate the role of the basal ganglia in the acquisition of categorical knowledge via an examination of the ability of people with Parkinson's disease (PD) to learn an artificial grammar (AG) system in a format akin to habit learning tasks such as probabilistic classification learning. Eighteen nondemented patients with PD were compared with 22 matched controls on a task modified so that participants' grammar learning depended on making use of feedback provided on each trial. Results showed that patients with PD exhibited abnormal AG learning, with the deficit more pronounced early in the task. Impaired categorisation performance in patients was not related to declarative knowledge obtained during the test and was independent of frontal functioning. The findings presented here indicate that prior inconsistencies across category learning tasks in PD performance may be explained, at least in part, by whether the task necessitates the formation of associations between stimulus cues and categorical responses, and support the hypothesis that the basal ganglia play a specific role in procedural learning in complex feedback-based categorisation tasks. PMID- 16359648 TI - Parametric effects of numerical distance on the intraparietal sulcus during passive viewing of rapid numerosity changes. AB - A number of functional neuroimaging studies have revealed that regions in and around the intraparietal sulcus (IPS) are parametrically modulated by numerical distance, whereby there is an inverse relationship between distance and levels of activation. These areas are thus thought to contain the internal representation of numerical magnitude. Nevertheless, it has also been suggested that the IPS is involved in response selection during number comparison tasks rather than the representation of numerical magnitude per se. In order to test the independence of the effect of distance on cortical regions, we employed a passive viewing paradigm. Sixteen right-handed male participants viewed rapidly changing slides containing arrays of squares. By varying the distance between the numerosity presented in separate blocks (8 vs. 8, 8 vs. 12, and 8 vs. 16), we examined which regions exhibit a parametric effect of numerical distance. This analysis revealed such effects in the superior part of the IPS bilaterally as well as the superior parietal lobule and the supramarginal gyrus. In contrast, slides rapidly changing in area but not number (Area constant, Area x 1, and Area x 2) did not yield a parametric effect of distance in these regions. Instead, a reverse effect of area was found in a region of the calcarine sulcus. These findings suggest that areas in and around the IPS are involved in numerical magnitude discrimination in the absence of an explicit task and response requirements. PMID- 16359649 TI - CA3 axonal sprouting in kainate-induced chronic epilepsy. AB - Latency between an early neurological insult and development of spontaneous recurrent seizures suggests aberrant chronological reorganization in patients with mesial temporal sclerosis associated epilepsy. Kainate-induced status similarly results in delayed development of spontaneous recurrent seizures. Mossy fiber sprouting by the dentate granule cells is a well-characterized manifestation of such temporal structural reorganization in both patients and animal models. However, alterations in other components of hippocampal circuitry have not been evaluated. We present results from studies using precise anterograde and retrograde tract tracing methodologies to evaluate the reorganization of outflow of the CA3 pyramidal cells. Although septotemporal relationships of the normal CA3 outflow tract through the Schaffer collaterals are well known, their aberrant reorganization following kainate-induced spontaneous recurrent seizures is not known. We provide the first definitive evidence of widespread CA3 structural reorganization in the form of sprouting of CA3 axons to widespread areas throughout the hippocampus and entorhinal cortex. This includes an apparent increase in the density of projection to areas that normally receive CA3 outflow such as CA1 and subiculum as well as novel projections beyond the confines of the hippocampus to the pre and parasubiculum and medial and lateral entorhinal cortex. We provide the first evidence of novel CA3 Schaffer collateral projection to the entorhinal cortex. The sprouting of CA3 outflow to widespread regions of the hippocampus and the entorhinal cortex may provide insight into how the injured hippocampus propagates unconventional impulse excitation to cortical fields which have a critical role in providing excitatory inputs into the hippocampus possibly setting up reverberating excitatory circuits as well as widespread connections throughout the cortical mantle. Sprouting-related mechanisms may also explain the latency associated with development of spontaneous recurrent seizures, the hallmark of temporal lobe epilepsy. PMID- 16359651 TI - Structure and assembly of epiglucan, the extracellular (1-->3;1-->6)-beta-glucan produced by the fungus Epicoccum nigrum strain F19. AB - In a previous article [Carbohydr. Res.2001, 331, 163-171] two different structures for the possible modular repeating unit of the extracellular beta glucan, epiglucan produced by the fungus Epicoccum nigrum strain F19 were proposed. Clarifying which was the more likely one was considered essential before attempts were made to understand how epiglucan was assembled by this fungus. Data from Smith degradation analyses of epiglucan were consistent with the repeating unit of structure I, where single glucosyl residues are attached by (1-->6)-beta-linkages to two out of every three glucosyl residues in the (1-->3) beta-linked glucan backbone. Repeated Smith degradations of 14C-glucose labelled epiglucan showed that chain elongation occurred from its non-reducing end. Side chain insertion into the growing glucan was followed by analysis of real time incorporation of 13C-glucose into epiglucan by 13C NMR, and 14C-glucose by enzymic digestion of the synthesised 14C-epiglucan. All data obtained were consistent with the view that single (1-->6)-beta-linked glucosyl side residues are inserted simultaneously as the glucan backbone elongates. PMID- 16359652 TI - Enumeration versus multiple object tracking: the case of action video game players. AB - Here, we demonstrate that action video game play enhances subjects' ability in two tasks thought to indicate the number of items that can be apprehended. Using an enumeration task, in which participants have to determine the number of quickly flashed squares, accuracy measures showed a near ceiling performance for low numerosities and a sharp drop in performance once a critical number of squares was reached. Importantly, this critical number was higher by about two items in video game players (VGPs) than in non-video game players (NVGPs). A following control study indicated that this improvement was not due to an enhanced ability to instantly apprehend the numerosity of the display, a process known as subitizing, but rather due to an enhancement in the slower more serial process of counting. To confirm that video game play facilitates the processing of multiple objects at once, we compared VGPs and NVGPs on the multiple object tracking task (MOT), which requires the allocation of attention to several items over time. VGPs were able to successfully track approximately two more items than NVGPs. Furthermore, NVGPs trained on an action video game established the causal effect of game playing in the enhanced performance on the two tasks. Together, these studies confirm the view that playing action video games enhances the number of objects that can be apprehended and suggest that this enhancement is mediated by changes in visual short-term memory skills. PMID- 16359653 TI - Differential developmental trajectories for egocentric, environmental and intrinsic frames of reference in spatial memory. AB - We studied the development of spatial frames of reference in children aged 3-6 years, who retrieved hidden toys from an array of identical containers bordered by landmarks under four conditions. By moving the child and/or the array between presentation and test, we varied the consistency of the hidden toy with (i) the body, and (ii) the testing room. The toy's position always remained consistent with (iii) the array and bordering landmarks. We found separate, additive performance advantages for consistency with body and room. These effects were already present at 3 years. A striking finding was that the room effect, which implies allocentric representations of the room and/or egocentric representations updated by self-motion, was much stronger in the youngest children than the body effect, which implies purely egocentric representations. Children as young as 3 years therefore had, and greatly favoured, spatial representations that were not purely egocentric. Viewpoint-independent recall based only on the array and bordering landmarks emerged at 5 years. There was no evidence that this later developing ability, which implies object-referenced (intrinsic) representations, depended on verbal encodings. These findings indicate that core components of adult spatial competence, including parallel egocentric and nonegocentric representations of space, are present as early as 3 years. These are supplemented by later-developing object-referenced representations. PMID- 16359654 TI - Two spatial memories are not better than one: evidence of exclusivity in memory for object location. AB - This paper studies the dynamics of attempting to access two spatial memories simultaneously and its implications for the accuracy of recall. Experiment 1 demonstrates in a range of conditions that two cues pointing to different experiences of the same object location produce little or no higher recall than that observed with a single cue. Experiment 2 confirms this finding in a within subject design where both cues have previously elicited recall. Experiment 3 shows that these findings are only consistent with a model in which two representations of the same object location are mutually exclusive at both encoding and retrieval, and inconsistent with models that assume information from both representations is available. We propose that these representations quantify directionally specific judgments of location relative to specific anchor points in the stimulus; a format that precludes the parallel processing of like representations. Finally, we consider the apparent paradox of how such representations might contribute to the acquisition of spatial knowledge from multiple experiences of the same stimuli. PMID- 16359655 TI - Carbohydrate partitioning between upper and lower regions of the crown in oat and rye during cold acclimation and freezing. AB - Carbohydrates have long been recognized as an important aspect of freezing tolerance in plants but the association between these two factors is often ambiguous. To help clarify the relationship, the allocation of carbohydrates between specific tissues within the over wintering organ (crown) of winter cereals was measured. A winter-hardy and non-winter-hardy oat (Avena sativa L.), and a rye (Secale cereale L.) cultivar were grown and frozen under controlled conditions. Crown tissue was fractionated into an upper portion, called the apical region, and a lower portion, called the lower crown. These tissues were ground in liquid N and extracted with water. Extracts were analyzed by HPLC for the simple sugars, sucrose, glucose, fructose, and for fructan of various size classes. After 3 weeks of cold acclimation at 3 degrees C, carbohydrates accounted for approximately 40% of the dry weight of oats and 60% of the dry weight of rye. The apical region, which is the tissue within the crown that acclimates to the greatest extent, was generally 10% higher in total carbohydrates than the lower crown. During a mild freeze, various carbohydrates were allocated differently between specific tissues in the three genotypes. When frozen, fructan generally decreased to a greater extent in the lower crown than in the apical region but sugars increased more in the apical region than in the lower crown. Results suggest that to understand how carbohydrates relate to freezing tolerance, regions of the crown that endure freezing stress differently should be compared. PMID- 16359656 TI - Involvement of skin barrier dysfunction in itch-related scratching in special diet-fed hairless mice. AB - HR-1 hairless mice fed with a special diet develop atopic-like dry skin, characterized by increased transepidermal water loss, and prolonged bouts of spontaneous scratching. In this study, the role of the skin barrier dysfunction in the prolongation of scratching was evaluated. Although the prolonged scratching was dose-dependently inhibited by opioid receptor antagonist naloxone, neither H(1) receptor antagonist, mepyramine, nor 5-HT(1/2) receptor antagonist, methysergide, affected it. Thus, the prolonged scratching could be itch-related response independent of histamine and serotonin. The application of petrolatum ointment on the skin temporarily alleviated the increase of transepidermal water loss for 60 min after treatment. Due to this alleviation in barrier dysfunction, the prolongation of scratching was significantly suppressed. However, when the barrier dysfunction relapsed, the scratching worsened. Taken together, a skin barrier dysfunction is associated with the itch-related response. PMID- 16359657 TI - Antiproliferative and antiangiogenic effects of 3-methylcholanthrene, an aryl hydrocarbon receptor agonist, in human umbilical vascular endothelial cells. AB - There is increasing interest in the effects of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD, dioxin) and polycyclic aromatic hydrocarbons on cardiovascular diseases. Their chemical structures are similar, although polycyclic aromatic hydrocarbons contain no chlorine as does TCDD. The biochemical mechanism of their action is mainly mediated by the aryl hydrocarbon receptor. In addition, oxidative stress also plays a role in the biological and toxic effects of these chemicals. In this study, we used an aryl hydrocarbon receptor agonist, 3 methylcholanthrene (3-MC), to investigate its effect on the proliferation and angiogenesis of human umbilical vascular endothelial cells. 3-MC suppressed DNA synthesis of human umbilical vascular endothelial cells as determined by [(3)H]thymidine incorporation in a concentration-dependent fashion and arrested cells at the G0/G1 phase of the cell cycle. Interestingly, the inhibition of DNA synthesis by 3-MC was eliminated to a greater extent by aryl hydrocarbon receptor antagonists, alpha-NF (0.5 and 1 microM) and resveratrol (5 and 10 microM), than by the antioxidant, N-acetylcysteine (5 and 10 mM). Cell permeability, adhesion, and tube formation in human umbilical vascular endothelial cells exposed to 3-MC decreased in concentration-dependent manners. We also demonstrated that cell adhesion signaling (phosphorylated focal adhesion kinase (FAK)) decreased upon 3 MC treatment, suggesting that cell adhesion inhibited by 3-MC might be due to inhibition of cell adhesion signaling. Additionally, alpha-naphthoflavon (alpha NF) ameliorated the effects of 3-MC on cell permeability, adhesion and tube formation, indicating the involvement of the aryl hydrocarbon receptor in angiogenesis. The results suggest that the adverse effects of 3-MC are mainly mediated by the aryl hydrocarbon receptor and not via increased oxidative stress. PMID- 16359658 TI - Natural products from ginseng inhibit [3H]batrachotoxinin A 20-alpha-benzoate binding to Na+ channels in mammalian brain. AB - A [(3)H]batrachotoxinin A-20alpha-benzoate ([(3)H]BTX-B) binding assay was used to investigate the interaction of two ginseng aglycones (20(S)protopanaxadiol and 20(S)protopanaxatriol) and Rh(2) (a monoglucoside of 20(S)protopanaxadiol) with voltage-gated sodium channels in mouse brain. All compounds inhibited the binding of [(3)H]BTX-B and IC(50)s were established at 42 microM (20(S)protopanaxadiol), 79 microM (20(S)protopanaxatriol) and 162 microM (Rh(2)). Scatchard analysis confirmed that 20(S)protopanaxadiol and Rh-2 reduced the B(max) of [(3)H]BTX-B binding while Rh(2) also increased the K(d). At IC(50) concentrations and above, 20(S)protopanaxadiol and Rh(2) increased the dissociation of the [(3)H]BTX B:sodium channel complex above that produced by a saturating concentration of veratridine, but failed to reduce the rate of association of [(3)H]BTX-B with sodium channels. Reversal of the inhibition of [(3)H]BTX-B binding by 20(S)protopanaxadiol and Rh(2) occurred slowly. We conclude that the 20(S)protopanaxadiol and the less potent inhibitor Rh(2) destabilize BTX-B activated sodium channels through non-covalent allosteric modification of neurotoxin binding site 2. PMID- 16359659 TI - Benfotiamine relieves inflammatory and neuropathic pain in rats. AB - Benfotiamine has shown therapeutic efficacy in the treatment of painful diabetic neuropathy in human beings. However, so far there is no evidence about the efficacy of this drug in preclinical models of pain. The purpose of this study was to assess the possible antinociceptive and antiallodynic effect of benfotiamine in inflammatory and neuropathic pain models in the rat. Inflammatory pain was induced by injection of formalin in non-diabetic and diabetic (2 weeks) rats. Reduction of flinching behavior was considered as antinociception. Neuropathic pain was induced by either ligation of left L5/L6 spinal nerves or administration of streptozotocin (50 mg/kg, i.p.) in Wistar rats. Benfotiamine significantly reduced inflammatory (10-300 mg/kg) and neuropathic (75-300 mg/kg) nociception in non-diabetic and diabetic rats. Results indicate that oral administration of benfotiamine is able to reduce tactile allodynia from different origin in the rat and they suggest the use of this drug to reduce inflammatory and neuropathic pain in humans. PMID- 16359660 TI - Potent pruritogenic action of tryptase mediated by PAR-2 receptor and its involvement in anti-pruritic effect of nafamostat mesilate in mice. AB - The pruritogenic potency of tryptase and its involvement in anti-pruritic effect of intravenous nafamostat mesilate (NFM) were studied in mice. An intradermal injection of tryptase (0.05-1 ng/site) elicited scratching in ICR mice, while chymase was without effects at doses of 0.05-50 ng/site. The dose-response curve of tryptase action was bell-shaped and the effect peaked at 0.1 ng/site (approximately 0.7 fmol/site). NFM (10 mg/kg) inhibited scratching induced by tryptase but not by histamine and serotonin. NFM (1-10 mg/kg) produced the dose dependent inhibition of scratching induced by intradermal compound 48/80 (10 microg/site). The inhibition by NFM (10 mg/kg) was abolished in mast cell deficient (WBB6F1 W/W(V)) mice, but not in wild-type (WBB6F1 +/+) mice. NFM (10 mg/kg) suppressed tryptase activity in the mouse skin. Proteinase-activated receptor-2 (PAR-2) neutralizing antibody (0.1 and 1 microg/site) and the PAR-2 antagonist FSLLRY (10 and 100 microg/site) inhibited scratching induced by tryptase (0.1 ng/site) and compound 48/80 (10 microg/site). These results suggest that mast cell tryptase elicits itch through PAR-2 receptor and that NFM inhibits itch-associated responses mainly through the inhibition of mast cell tryptase. PMID- 16359661 TI - Clinical and microstructural analysis of patients with hyper-reflective corneal endothelial nuclei imaged by in vivo confocal microscopy. AB - The purpose of this study was to determine the significance of hyper-reflective corneal endothelial nuclei imaged by in vivo confocal microscopy. A retrospective analysis was performed using a database of 505 patients that had undergone in vivo confocal microscopy of the cornea. All subjects with hyper-reflective endothelial nuclei were identified and these images were analysed to determine corneal endothelial cell density and morphology. The clinical notes of these patients were reviewed and corresponding data regarding corneal thickness was obtained from a related database of Orbscan II pachymetry. Hyper-reflective endothelial nuclei were identified in 41 eyes of 39 (7.7%) patients. Diagnoses included previous cataract surgery or penetrating keratoplasty, posterior polymorphous dystrophy, Fuchs' endothelial dystrophy and irido-corneal endothelial syndrome. No patients with clinically normal corneas exhibited bright endothelial nuclei. The mean endothelial cell density in this group was 1325+/ 872 cells mm(-2) and endothelial density was below age-adjusted normal values in 69.2% of patients. Both cellular polymegathism (coefficient of variation of cell area 33.9+/-7.4%) and cellular pleomorphism were noted (51.8+/-9.0% hexagonal cells). The mean central corneal thickness was 582+/-52 microm. There was no significant difference in endothelial density and morphology compared to cases that had low endothelial density but did not exhibit bright nuclei. In conclusion, this study is the first to investigate the significance of bright endothelial nuclei detected by in vivo confocal microscopy. The strong association with corneal disease states suggests that the most likely explanation for this appearance is the alteration in cellular/nuclear morphology, composition or function. PMID- 16359662 TI - Inhibition of retinal neovascularization by soluble EphA2 receptor. AB - Eph receptor tyrosine kinases (RTKs) and their ligands, known as ephrins, play an important role in vascular remodeling during embryogenesis, but their functions in adult angiogenesis are just beginning to be investigated. In this report, we investigated the effect of blocking EphA receptor activation on VEGF-induced angiogenic responses of cultured retinal endothelial cells and on retinal neovascularization in a rodent model of retinopathy of prematurity (ROP). Soluble EphA2-Fc receptors inhibited ephrin-A1 ligand or VEGF-induced BRMEC migration and tube formation without affecting proliferation in vitro. Since EphA2-Fc receptors can inhibit activation of multiple EphA receptors, the specific role of EphA2 receptor in angiogenesis was further investigated in EphA2-deficient endothelial cells. Loss of EphA2 in endothelial cells leads to defective cell migration and assembly in response to either ephrin-A1 or VEGF. Finally, a significant reduction in the severity of abnormal retinal neovascularization was observed in the eyes treated with soluble EphA2-Fc receptors, yet the normal total retinal vascular area was not significantly changed. Because soluble Eph receptor significantly inhibited pathologic retinal angiogenesis without affecting normal intraretinal vessels, it may be a promising agent for treatment of retinal angiogenesis in a number of human ocular diseases. PMID- 16359663 TI - Posterior capsule opacification-like changes in rat lens explants cultured with TGFbeta and FGF: effects of cell coverage and regional differences. AB - Following cataract surgery, many patients suffer secondary loss of vision because of posterior capsule opacification (PCO), which arises when residual lens epithelial cells become aberrant and migrate into the light path. Transforming growth factor-beta (TGFbeta)-induced transdifferentiation of lens cells appears to play a key role in this process. Fibroblast growth factor (FGF) may also play a role by promoting the survival of TGFbeta-affected cells and influencing their subsequent behaviour. In the present study, the effects of two different TGFbeta and FGF treatment regimes were compared in rat lens epithelial explants with either low or high initial cell coverage. Explants treated with 50 pg ml(-1) TGFbeta2 and 20 ng ml(-1) FGF-2 sequentially (day 0, day 1) or simultaneously (day 0), then cultured for up to 30 days with FGF, were assessed by light microscopy and immunolocalisation of markers for transdifferentiation (alpha smooth muscle actin (alphaSMA) and type I collagen) or lens epithelial phenotype (Pax6) and fibre differentiation (beta-crystallin). By day 4, most cells had lost Pax6 reactivity, alphaSMA reactivity was evident, and there were differences between growth factor treatment groups, low and high initial cell coverage explants, and peripheral and central regions of explants. On day 30 of culture, all explants were well populated with cells, irrespective of treatment and initial cell coverage, and exhibited diverse PCO-like morphological changes, with expression of transdifferentiation markers and beta-crystallin in virtually all cells. Such overall resilience to variations in conditions may contribute to the insidious nature of PCO, while factors related to observed early differences between groups may contribute to PCO pleiomorphism. PMID- 16359664 TI - Complex neurodegeneration in retina following moderate ischemia induced by bilateral common carotid artery occlusion in Wistar rats. AB - Bilateral common carotid artery occlusion (BCCAO) produces moderate levels of ischemia in the retina of rats, which may simulate the inflow disturbances in severe carotid artery disease. ERG changes following acute BCCAO have been well described, but the effects of chronic BCCAO on the histopathology of the retina remain to be characterized in a reproducible model. Chronic BCCAO was induced in halothane-anaesthetized male Wistar rats and the retina fixed after 3, 6, or 24 hr, 1 week, and 2, 4, or 6 months. Cell counts and measurements of retinal layers were performed in H&E stained paraffin sections. Immunohistochemistry with a panel of fourteen antibodies served to examine the survival of different retinal cell class, astrocytic reactions and the expression of acute stress response proteins. A lectin method was used to label activated microglial cells. Microglial activation, heme oxygenase-1 upregulation and caspase-3 cleavage occurred during the first 24hr in the absence of overt cell death of retinal ganglion cells (RGC). Three waves of neurodegeneration followed. RGCs were affected after 1 week, followed by neurons in the inner nuclear layer at 2 months, and finally photoreceptors at 4 months. Immunomarkers indicated acute damage to horizontal cells and prolonged survival of amacrine cells. In conclusion, chronic BCCAO produced delayed neuronal death in the retina of adult male Wistar rats. The window of moderate changes of at least 1 day may facilitate molecular studies on retinal ganglion cell loss. PMID- 16359665 TI - Modulation of the antigen transport machinery TAP by friends and enemies. AB - The transporter associated with antigen processing (TAP) is a key factor of the major histocompatibility complex (MHC) class I antigen presentation pathway. This ABC transporter translocates peptides derived mainly from proteasomal degradation from the cytosol into the ER lumen for loading onto MHC class I molecules. Manifold mechanisms have evolved to regulate TAP activity. During infection, TAP expression is upregulated by interferon-gamma. Furthermore, the assembly and stability of the transport complex is promoted by various auxiliary factors. However, tumors and viruses have developed sophisticated strategies to escape the immune surveillance by suppressing TAP function. The activity of TAP can be impaired on the transcriptional or translational level, by enhanced degradation or by inhibition of peptide translocation. In this review, we briefly summarize existing data concerning the regulation of the TAP complex. PMID- 16359666 TI - Common antigenicity for two glycosidases. AB - Enzyme replacement therapy (ERT) has proven to be an effective therapy for some lysosomal storage disorder (LSD) patients. A potential complication during ERT is the generation of an immune response against the replacement protein. We have investigated the antigenicity of two distantly related glycosidases, alpha glucosidase (Pompe disease or glycogen storage disease type II, GSD II), and alpha-L-iduronidase (Hurler syndrome, mucopolysaccharidosis type I, MPS I). The linear sequence epitope reactivity of affinity purified polyclonal antibodies to recombinant human alpha-glucosidase and alpha-L-iduronidase was defined, to both glycosidases. The polyclonal antibodies exhibited some cross-reactive epitopes on the two proteins. Moreover, a monoclonal antibody to the active site of alpha glucosidase showed cross-reactivity with a catalytic structural element of alpha L-iduronidase. In a previous study, in MPS I patients who developed an immune response to ERT, this same site on alpha-L-iduronidase was highly antigenic and the last to tolerise following repeated enzyme infusions. We conclude that glycosidases can exhibit cross-reactive epitopes, and infer that this may relate to common structural elements associated with their active sites. PMID- 16359668 TI - Molecular dynamics simulation of a phosphatidylglycerol membrane. AB - Although molecular dynamics simulations are an important tool for studying membrane systems, relatively few simulations have used anionic lipids. This paper reports the first simulation of a pure phosphatidylglycerol (PG) bilayer. The properties of this equilibrated palmitoyloleoylphosphatidylglycerol membrane agree with experimental observations of PG membranes and with previous simulations of monolayers and mixed bilayers containing PG lipids. These simulations also provide interesting insights into hydrogen bonding interactions in PG membranes. This equilibrated membrane will be a useful starting point for simulations of membrane proteins interacting with PG lipids. PMID- 16359667 TI - The ABC of auxin transport: the role of p-glycoproteins in plant development. AB - A surprising outcome of the Arabidopsis genome project was the annotation of a large number of sequences encoding members of the ABC transporter superfamily, including 22 genes encoding the p-glycoprotein (PGP) subfamily. As mammalian PGP orthologs are associated with multiple drug resistance, plant PGPs were initially presumed to function in detoxification, but were soon seen to have a developmental role. Here, we summarise recent studies of plant PGPs indicating that PGPs mediate the cellular and long-distance transport of the plant hormone auxin. One class of PGPs, represented by AtPGP1, catalyze auxin export, while another class with at least one member, AtPGP4, appears to function in auxin import. Current models on the physiological role of PGPs, their functional interaction and their involvement in cell-to cell (polar) auxin transport are discussed. PMID- 16359669 TI - Isolation and functional characterization of fatty acid delta5-elongase gene from the liverwort Marchantia polymorpha L. AB - Bryophyte Marchantia polymorpha L. produces C22 very-long-chain polyunsaturated fatty acid (VLCPUFA). Thus far, no enzyme that mediates elongation of C20 VLCPUFAs has been identified in land plants. Here, we report the isolation and characterization of the gene MpELO2, which encodes an ELO-like fatty acid elongase in M. polymorpha. Heterologous expression in yeast demonstrated that MpELO2 encodes delta5-elongase, which mediates elongation of arachidonic (20:4) and eicosapentaenoic acids (20:5). Phylogenetic and gene structural analysis indicated that the MpELO2 gene is closely related to bryophyte Delta6-elongase genes for C18 fatty acid elongation and diverged from them by local gene duplication. PMID- 16359670 TI - Differences in catalytic activity between rat testicular and ovarian carbonyl reductases are due to two amino acids. AB - The sequences of rat testis carbonyl reductase (rCR1) and rat ovary carbonyl reductase (rCR2) are 98% identical, differing only at amino acids 140, 141, 143, 235 and 238. Despite such strong sequence identity, we find that rCR1 and rCR2 have different catalytic constants for metabolism of menadione and 4-benzoyl pyridine. Compared to rCR1, rCR2 has a 20-fold lower K(m) and 5-fold lower k(cat) towards menadione and a 7-fold lower K(m) and 7-fold lower k(cat) towards 4 benzoyl-pyridine. We constructed hybrids of rCR1 and rCR2 that were changed at either residues 140, 141 and 143 or residues 235 and 238. rCR1 with residues 140, 141 and 143 of rCR2 has similar catalytic efficiency for menadione and 4-benzoyl pyridine as rCR1. rCR1 with Thr-235 and Glu-238 of rCR2 has the catalytic constants of rCR2, indicating that it is this part of rCR2 that contributes to its lower K(m) for menadione and 4-benzoyl-pyridine. Comparisons of three dimensional models of rCR1 and rCR2 show how Thr-235 and Glu-238 stabilize rCR2 binding of NADPH and menadione. PMID- 16359671 TI - Differential beta-arrestin binding of AT1 and AT2 angiotensin receptors. AB - Agonist stimulation of G protein-coupled receptors causes receptor activation, phosphorylation, beta-arrestin binding and receptor internalization. Angiotensin II (AngII) causes rapid internalization of the AT1 receptors, whereas AngII-bound AT2 receptors do not internalize. Although the activation of the rat AT1A receptor with AngII causes translocation of beta-arrestin2 to the receptor, no association of this molecule with the AT2 receptor can be detected after AngII treatment with confocal microscopy or bioluminescence resonance energy transfer. These data demonstrate that the two subtypes of angiotensin receptors have different mechanisms of regulation. PMID- 16359672 TI - Hansenula polymorpha and Saccharomyces cerevisiae Pex5p's recognize different, independent peroxisomal targeting signals in alcohol oxidase. AB - Peroxisomal alcohol oxidase (AO) from Hansenula polymorpha is inactive and partially mislocalized to the cytosol upon synthesis in Saccharomyces cerevisiae. Co-production with H. polymorpha pyruvate carboxylase (HpPyc1p) resulted in AO activation, but did not improve import into peroxisomes. We show that import of AO mediated by S. cerevisiae Pex5p is strictly dependent on the peroxisomal targeting signal 1 (PTS1) of AO and independent of HpPyc1p. In contrast, HpPex5p mediated sorting of AO into S. cerevisiae peroxisomes is independent of the PTS1, but requires an alternative PTS that is only formed when HpPyc1p is co-produced and most likely involves folding and co-factor binding to AO. PMID- 16359673 TI - Involvement of de novo ceramide biosynthesis in macrophage death induced by activation of ATP-sensitive P2X7 receptor. AB - Macrophage ionotropic P2X7 receptors regulate cell-death through ill-defined signaling pathways. Here, we investigated the role of ceramide, an apoptogenic sphingolipid and showed that ATP stimulated ceramide accumulation in macrophages. Benzoylbenzoyl-ATP, a potent P2X7 agonist, was able to mimic the effects of ATP on ceramide accumulation while oxidized ATP had the opposite effect. Ceramide accumulation was blocked by de novo ceramide biosynthesis inhibitors. Interestingly, ATP-induced caspase-3/7 activation was dependent on ceramide generation. Finally, we showed that de novo ceramide biosynthesis is involved in ATP-induced macrophage death in a caspase-dependent manner. Our results indicate a novel role of ceramide in P2X7-regulated cell-death. PMID- 16359674 TI - Osmotic stress sensing in Populus: components identification of a phosphorelay system. AB - To study the Populus response to an osmotic stress, we have isolated one cDNA encoding a histidine-aspartate kinase (HK1) and four cDNAs encoding histidine containing phosphotransfer proteins (HPts), HPt1-4. The predicted HK1 protein shares a typical structure with ATHK1 and SLN1 osmosensors. The 4 HPTs are characterized by the histidine phosphotransfer domain. We have shown that HK1 is upregulated during an osmotic stress in hydroponic culture. We have detected an interaction between HK1 and HPt2, using the yeast two-hybrid system. These results suggest the existence of a multi-step phosphorelay pathway probably involved in osmotic stress sensing in Populus. PMID- 16359675 TI - Exploring the capacity of trigger factor to function as a shield for ribosome bound polypeptide chains. AB - Ribosome-bound trigger factor (TF) is the first chaperone encountered by a nascent polypeptide chain in bacteria. TF has been proposed to form a cradle shaped shield for nascent chains up to approximately 130 residues to fold in a protected environment upon exit from the ribosome. We report that nascent chains of luciferase up to 280 residues in length are relatively protected by TF against digestion by proteinase K. In contrast, nascent chains of the constitutively unstructured protein alpha-synuclein were not protected, although they were in close proximity to TF by crosslinking. Thus, TF is not a general shield for nascent chains. Protease protection appears to depend on a hydrophobic interaction of TF with nascent polypeptides. PMID- 16359676 TI - Comparison of synthesis of 15 alpha-hydroxylated steroids in males of four North American lamprey species. AB - Recent studies have provided evidence that 15 alpha-hydroxytestosterone (15 alpha T) and 15 alpha-hydroxyprogesterone (15 alpha-P) are produced in vitro and in vivo in adult male sea lampreys (Petromyzon marinus), and that circulatory levels increase in response to injections with gonadotropin-releasing hormone (GnRH). We examined four species from the Petromyzontidae family including silver lampreys (Ichthyomyzon unicuspis), chestnut lampreys (I. castaneus), American brook lampreys (Lethenteron appendix), and Pacific lampreys (Entosphenus tridentatus) to determine if these unusual steroids were unique to sea lampreys or a common feature in lamprey species. In vitro production was examined through incubations of testis with tritiated precursors, and 15 alpha-T and 15 alpha-P production was confirmed in all species through co-elution with standards on both high performance liquid chromatography (HPLC) and thin layer chromatography. In vivo production was proven by demonstrating that HPLC-fractionated plasma had peaks of immunoreactive 15 alpha-T and 15 alpha-P that co-eluted with standards through using previously developed radioimmunoassays for 15 alpha-T and 15 alpha-P. The possible functionality of 15 alpha-T and 15 alpha-P was further examined in silver and Pacific lampreys by investigating the effect of injection of either type of lamprey GnRH on plasma concentrations of 15 alpha-T and 15 alpha-P. Injections with exogenous GnRH did not affect circulatory levels of either steroid in silver lampreys, and only GnRH III elicited higher levels of both steroids in Pacific lampreys. The 15 alpha-hydroxylase enzyme(s) for steroids appeared to present in adult males of all species examined, but the question of whether 15 alpha-hydroxylated steroids are functional in these lamprey species, and the significance of the 15-hydroxyl group, requires further research. PMID- 16359677 TI - Perceptions and practice of concordance in nurses' prescribing consultations: findings from a national questionnaire survey and case studies of practice in England. AB - BACKGROUND: The number of nurses able to independently prescribe medicines in England is increasing. Patient adherence to prescribed medicines remains a significant problem [Department of Health, 2000. Pharmacy in the Future: Implementing the NHS Plan. A Programme for Pharmacy in the NHS. Stationary Office, London]. Concordance-a partnership approach to medicine consultations-is advocated as an effective solution [Medicines Partnership, 2003. Project Evaluation Toolkit. Medicines Partnership, London]. OBJECTIVES: To investigate whether nurses were practising the principles of concordance within their prescribing interactions. DESIGN: Phase (i) postal questionnaire survey. Phase (ii): case studies of practice. SETTINGS: Phase (i) primary and secondary care trusts throughout England in which nurse prescribers were practicing. Phase (ii) six general practice settings; one community midwifery service; one specialist community palliative care service; one secondary care ophthalmology unit; one NHS walk-in centre. PARTICIPANTS: Phase (i) a random sample of 246 nurses registered as independent nurse prescribers with the Nursing and Midwifery Council in 2002/2003. Phase (ii) purposively selected sample of 14 nurse prescribers who participated in Phase (i) of the study; a total of 208 purposively selected patients completed self-administered questionnaires. METHODS: Phase (i) postal questionnaires. Phase (ii) structured non-participant observation of 118 nurse prescribing consultations; 115 post-consultation patient questionnaires; 93 patient postal questionnaires. RESULTS: 99% of the nurses in the national survey stated they were practising the principles of concordance. The majority of patients surveyed also reported experiencing concordance in practice. Observation of practice revealed that although some principles of concordance were regularly integrated into nurses' practice, other principles were less often in evidence. Some evidence from both observation of practice and patient questionnaires suggested that a professionally determined 'compliance' agenda may still be partially operating in practice. CONCLUSIONS: Most nurses believe they are practicing concordance in their prescribing consultations. The majority of patients also reported that they had experienced some of the principles of concordance in practice. Observation of practice highlighted that the shift from a professionally determined compliance agenda to the integration of concordance into nurses' prescribing consultations had not yet taken place. PMID- 16359678 TI - Microarray analysis identifies genes preferentially expressed in the lung schistosomulum of Schistosoma mansoni. AB - The lung schistosomulum of Schistosoma mansoni is a validated target of protective immunity elicited in vaccinated mice. To identify genes expressed at this stage we constructed a microarray, representing 3088 contigs and singlets, with cDNA derived from in vitro cultured larvae and used it to screen RNA from seven life-cycle stages. Clustering of genes by expression profile across the life cycle revealed a number of membrane, membrane-associated and secreted proteins up-regulated at the lung stage, that may represent potential immune targets. Two promising secreted molecules have homology to antigens with vaccine and/or immunomodulatory potential in other helminths. PMID- 16359679 TI - Development of photochemical method for meniscal repair: a preliminary study. AB - Visible light combined with naphthalimides has previously been shown to catalyze formation of physical bonds in avascular meniscal tissue. The first objective was to modify the existing in vitro testing method (i.e., adhesion testing using lap jointed slices) to gain more sensitivity in detecting relative bonding strengths among candidate bonding agents. A repeated measures experimental design (RMED) was used to account for variability in properties among bovine menisci and was achieved by testing all treatments/controls on slices from each meniscus. Additionally, to make the method more clinically relevant in modeling a bucket handle tear, the bovine meniscal slices were cut with collagen fibers parallel to the test slice's length. Peak stress was greater for the complete treatment group (light plus naphthalimide) than for the control or incomplete treatment groups (light only or napthalimide only). The second objective was to perform concentration and photoactivation time dose-response studies. In the concentration dose-response study, peak stress was greater for all treatments when compared with the control but not different among treatment groups; however, there was a trend of increased bonding strength with increased concentration. In the photoactivation time dose-response study, peak stress was greater for all treatments when compared with the control and greater for the 3-min treatment vs. the 6- and 9-min treatments. Peak stress was not different between the longer treatments. The RMED provided increased reproducibility and statistical sensitivity for detecting differences among treatments and will be used to test candidate bonding agents prior to in vivo testing. PMID- 16359680 TI - Comparison of micro-level and continuum-level voxel models of the proximal femur. AB - Continuum-level finite element (FE) models became standard computational tools for the evaluation of bone mechanical behavior from in vivo computed tomography scans. Such scans do not account for the anisotropy of the bone. Instead, local mechanical properties in the continuum-level FE models are assumed isotropic and are derived from bone density, using statistical relationships. Micro-FE models, on the other hand, incorporate the anisotropic structure in detail. This study aimed to quantify the effects of assumed isotropy, by comparing continuum-level voxel models of a healthy and a severely osteoporotic proximal femur with recently analyzed micro-FE models of the same bones. The micro-model element size was coarsened to generate continuum FE models with two different element sizes (0.64 and 3.04 mm) and two different density-modulus relationships found in the literature for wet and ash density. All FE models were subjected to the same boundary conditions that simulated a fall to the side, and the stress and strain distributions, model stiffness and yield load were compared. The results indicated that the stress and strain distributions could be reproduced well with the continuum models. The smallest differences between the continuum-level model and micro-level model predictions of the stiffness and yield load were obtained with the coarsest element size. Better results were obtained for both continuum element sizes when isotropic moduli were based on ash density rather than wet density. PMID- 16359681 TI - Analysis of the essential oil composition of eight Anthemis species from Greece. AB - The volatile composition of eight Anthemis species has been studied. The essential oils were obtained by hydrodistillation in a modified Clevenger-type apparatus, and their analyses were performed by GC and GC-MS. Identification of the substances was made by comparison of mass spectra and retention indices with literature records. A total of 284 different compounds were identified and significant qualitative and quantitative differences and similarities were observed among the samples. The main constituents of the investigated populations of each taxon have been revealed as follows: A. altissima: (-)-linalool, trans caryophyllene, cis-chrysanthenyl acetate; A. auriculata: spathulenol, trans caryophyllene, beta-eudesmol; A. chia: cis-chrysanthenyl acetate, trans caryophyllene, germacrene-d; A. cotula: germacrene-d, spathulenol A. tinctoria: spathulenol, (-)-caryophyllene oxide, T-cadinol; A. melanolepis: p-cymene, chrysanthenone, trans-verbenol, (-)-caryophyllene oxide; A. tomentosa: (-) linalool, 1,8-cineole; A. werneri subsp. werneri: nopol, terpineol-4, trans caryophyllene. Sesquiterpene hydrocarbons were shown to be the main group of constituents of all taxa. PMID- 16359682 TI - Monolithic beds of artificial gel antibodies. AB - The introductions of the continuous beds, now often called monoliths [S. Hjerten, J.-L. Liao, R. Zhang, J. Chromatogr. 473 (1989), 273-275] and the artificial, highly selective gel antibodies against antigens as large as proteins, viruses and cells [J.-L. Liao, Y. Wang, S. Hjerten, Chromatographia 42 (1996), 259-262] were breakthroughs in the design of chromatographic beds. This paper deals with a combination of these two methods, i.e., the artificial gel antibodies have been synthesized in the monolithic mode. As antigen we have used human hemoglobin. A comparison of the ion-exchange chromatograms of the eluates from the monolithic columns shows that the monolith prepared in the presence of hemoglobin adsorbed this protein, but not the other proteins in the sample (ribonuclease A and cytochrome c), i.e., this monolith was selective for hemoglobin, whereas the blank column (prepared in the absence of hemoglobin) had no selective properties, since none of the applied proteins were adsorbed. The diameter of the column was 6mm, but the same approach to synthesize a monolithic selective bed can very likely also be used for capillaries and microchips. PMID- 16359683 TI - Use of polyoxyethylene surfactants for the extraction of organochlorine pesticides from agricultural soils. AB - Two non-ionic surfactant mixtures (POLE and Polyoxyethylene 10 Cetyl ether, POLE and Polyoxyethylene 10 Stearyl ether) have been used for microwave-assisted extraction of six organochlorine pesticides from agricultural soils prior to being determined by HPLC-UV. An experimental design was applied for the determination of variables which affect to recovery and to optimize the extraction parameters, surfactant concentration and volume, microwave time and power. Under the optimized conditions, the method was applied to different soil samples in order to analyze the influence of soil characteristics on the pesticides extraction. The results obtained indicate that most of these compounds can be recovered in good yields with RSD lower than 10% and detection limit ranged between 80 and 800 ng g(-1) for the pesticides studied. The proposed method was applied to a reference soil sample and to aged soils. PMID- 16359684 TI - Development of a two-color laser fluorescence detector. On-line detection of internal standards and unknowns by capillary electrophoresis within the same sample. AB - A two-laser, two-color detector has been developed for the simultaneous detection of naturally occurring and recombinant (internal standards) cytokines within the same biological sample. The internal standards were labeled with Bimane and detected with a 408 nm laser while the natural cytokines were labeled with AlexaFluor633 and detected with a 633 nm laser. The two resulting electropherograms were plotted as overlaid traces and quantification of the natural materials determined by comparison with the standards. Using this system, recovery of all four cytokine standards was greater than 94% in both saline and cytokine-depleted plasma. These recoveries could be achieved with intra- and inter-assay coefficients of variance (c.v.) of less than 4.5 and 5.6, respectively. Application of this system to the examination of clinical samples demonstrated that measurement of the four pro-inflammatory cytokines could distinguish between normals, sub-clinical and clinical inflammation. An advantage of this approach is that direct calculation of unknowns by comparison to identical internal standards can shorten analytical time by eliminating the need for additional standard or calibration runs. PMID- 16359685 TI - Metallothionein quantification in clams by reversed-phase high-performance liquid chromatography coupled to fluorescence detection after monobromobimane derivatization. AB - In this paper, we describe a highly specific, sensitive and reliable method for total metallothionein (MT) quantification by RP-HPLC coupled to fluorescence detection following reaction with monobromobimane of thiols from metal-depleted MT after heat-denaturation of extracts in the presence of sodium dodecyl sulphate (SDS). SDS-polyacrylamide gel electrophoresis (SDS-PAGE) confirmed the identity of the peak resolved (t(R)=16.44) with MT: a highly fluorescent protein of approximately 8.3 kDa, in agreement with the high thiol content and low MT size. Other heat-resistant and Cys-containing proteins of 35 kDa were efficiently separated. The new method was successfully used to quantify MT content in digestive gland of clams from southern Spanish coastal sites with different metal levels, and is proposed as a tool for using MTs as biomarker in monitoring programmes. PMID- 16359686 TI - Quinolizidine alkaloids and phomopsins in lupin seeds and lupin containing food. AB - In recent years there has been growing interest in replacing (genetically modified) soya by lupin. Lupin seeds, flours and lupin containing food have been analyzed in order to assess the relevance of a potential health hazard given by mycotoxins and/or naturally occurring alkaloids. Since not all important alkaloids used for quantitation were commercially available, isolation of lupanine, 13alpha-hydroxylupanine and angustifoline from lupin flours of high alkaloid contents was performed. Alkaloids were analyzed by GC-MS/GC-FID in parallel, while the phomopsin mycotoxins were analyzed by ELISA, since chromatographic methods were not sensitive enough and required time-consuming sample cleanup. The analyzed lupin containing foods were free of phomopsins. In foods where lupin was only a minor constituent the alkaloid content was of no concern. However, roasted lupin beans intended as coffee surrogate had alkaloid contents close to the Australian intervention limit of 200 microg/g. PMID- 16359687 TI - Optimizing the integrated pulsed amperometric multicycle step waveform for the determination of tetracyclines. AB - A method of modified integrated pulsed amperometric detection with multicycle step waveform (Multi-IPAD) following high-performance liquid chromatography (HPLC) was applied for the determination of tetracyclines (TCs) including dimethyltetracycline (DMTC), oxytetracycline (OTC) and tetracycline (TC). The key advantages of the Multi-IPAD are the abilities to enhance sensitivity and reproducibility and the ability to keep working electrode clean through the use of a high-frequent waveform alteration in integration step and the use of a cleaning potential, which is quite different from conventional three-step potential waveform. The analyses were carried out using the mobile phase of acetonitrile-water mixture solution (10:90, v/v) containing 1% perchloric acid on a C(18) column at a flow rate of 0.21 mL/min. The IPAD waveform parameters were optimized to maximize the signal-to-noise ratio (S/N) and successfully applied for the sensitive detection of TCs. The detection limits (S/N=3, 20 microL injected) were 0.07 mg/L for DMTC, 0.08 mg/L for OTC and 0.05 mg/L for TC. The peak height relative standard deviations (RSDs) of every compound for replicate injection (n=15) determined were below 4.6%. PMID- 16359688 TI - Single-cell analysis by intracellular immuno-reaction and capillary electrophoresis with laser-induced fluorescence detection. AB - A novel method for single-cell analysis was developed by combining electroporation for intracellular immuno-reaction and capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection. Human interferon-gamma (IFN gamma) in natural killer (NK) cells was chosen as the test antigen. Two forms of IFN-gamma in single cells could be well separated and detected with a limit of detection of zeptomole. In this assay, the anti-IFN-gamma monoclonal antibody labeled with fluorescein isothiocyanate (Ab*) was introduced into NK cells by electrophoration for intracellular immuno-reaction. After completion of the intracellular immuno-reaction, the NK cells were chemically pre-perforated with digitonin to lyse easily. Then, one NK cell containing the complexes of IFN-gamma isoantigens with Ab* was electrokinetically injected into the capillary. The cell adsorbed on the tip of capillary was lysed by ultrasonication. Finally, the complexes of the different forms of IFN-gamma in the cell were separated and detected by CE-LIF detection. PMID- 16359689 TI - High-performance liquid chromatographic enantioseparation of alpha-substituted glycine analogs on a quinine-based anion-exchanger chiral stationary phase under variable temperature conditions. AB - The retention of enantiomers of chiral analytes, i.e. alpha-substituted glycine analogs, on a quinine-based anion-exchanger chiral stationary phase was studied in the temperature range of 5-70 degrees C and at different mobile phase compositions, using isocratic elution in the reversed-phase mode. By variation of both mobile phase composition and temperature, baseline separations could be achieved for these enantiomers. Separation could be optimized more quickly by adjusting the column temperature rather than the mobile phase composition. The dependence of the natural logarithms of retention and selectivity factors (lnk' and lnalpha) on the inverse of temperature, 1/T (van't Hoff plots) was used to determine thermodynamic data on the enantiomers. Calculated thermodynamic constants (Delta(DeltaH degrees ), Delta(DeltaS degrees ) and Delta(DeltaG degrees )) were applied to promote an understanding of the thermodynamic driving forces for retention in this chromatographic system. The elution sequence of the enantiomers in most cases was determined. PMID- 16359690 TI - Hyphenated chromatographic techniques for the rapid screening and identification of antioxidants in methanolic extracts of pharmaceutically used plants. AB - Phytochemical analysis is an important scientific research area, which normally relies on a number of rather laborious and time-consuming techniques for compound identification. Isolation of the ingredients of plant extracts in adequate quantities for spectral and biological analysis was the basis of this research. In this paper the possibility of on-line rapid screening of antioxidant components in methanolic plant extracts and their subsequent identification is reported. Based exclusively on hyphenated chromatographic techniques the methanolic extracts of Tilia europea, Urtica dioica, Lonicera periclymenum and Hypericum perforatum are initially screened for their antioxidant components via an on-line DPPH and ABTS radical scavenging technique. Structural elucidation of the active analytes is achieved by means of LC-MS and LC-UV-SPE-NMR. After the determination of the appropriate LC gradient, a minimal number of chromatographic runs with these hyphenated techniques are adequate for the acquisition of the necessary data, leading to the identification of the targeted compounds. Based on their UV, NMR and MS spectra, the antioxidant compounds identified in the extracts under study were found to be either flavonoid glycosides or mono- and dicaffeoylquinic acids. Although the aim of the study was to show the great potential of the LC-UV-NMR-DPPH/ABTS approach for the rapid screening and identification of plant constituents, the results produced in the course of this study also have some merit by themselves. Some of the compounds detected are reported for the first time in the specific plant extracts. PMID- 16359691 TI - A chromatographic estimate of the degree of surface heterogeneity of RPLC packing materials. III. Endcapped amido-embedded reversed phase. AB - The difference in adsorption behavior between a conventional monomeric endcapped C18 stationary phase (3.43 micromol/m2) and an endcapped polymeric RP-Amide phase (3.31 micromol/m2) was investigated. The adsorption isotherms of four compounds (phenol, caffeine, sodium 2-naphthalene sulfonate, and propranololium chloride) were measured by frontal analysis (FA) and the degree of heterogeneity of each phase for each solute was characterized by their adsorption energy distributions (AED), derived using the Expectation-Maximization method. The results show that only certain analytes (phenol and 2-naphthalene sulfonate) are sensitive to the presence of the polar embedded amide groups within the RP phase. Their binding constants on the amide-bonded phase are significantly higher than on conventional RPLC phases. Furthermore, an additional type of adsorption sites was observed for these two compounds. However, these sites having a low density, their presence does not affect much the retention factors of the two analytes. On the other hand, the adsorption behavior of the other two analytes (caffeine and propranololium chloride) is almost unaffected by the presence of the amide group in the bonded layer. Strong selective interactions may explain these observations. For example, hydrogen-bond interactions between an analyte (e.g., phenol or naphthalene sulfonate) and the carbonyl group (acceptor) or the nitrogen (donor) of the amido-embedded group may take place. No such interactions may take place with either caffeine or the cation propranololium chloride. This study confirms the hypothesis that analytes have ready access to locations deep inside the bonded layer, where the amide groups are present. PMID- 16359692 TI - Influence of vinyl monomers and temperature on immobilization of cellulose 3,5 dimethylphenylcarbamate onto silica gel as chiral stationary phases for high performance liquid chromatography. AB - Cellulose 3,5-dimethylphenylcarbamates bearing a low content of a vinyl group at the 6-position on the glucose units were synthesized by a previously developed regioselective method and chemically immobilized onto a vinylized silica gel as chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC). The immobilization of the derivatives was performed through a radical polymerization reaction with AIBN as the initiator in the presence of toluene. The effects of vinyl monomers, such as isoprene, 2,3-dimethyl-1,3-butadiene (DMBD), ethylene glycol dimethacrylate (EDMA) and 1,5-hexadiene, on the immobilization and enantioselectivities of the derivatives were investigated. The effect of the temperature used for the radical polymerization reaction on the immobilization was also examined. In addition, the direct comparison of the chiral recognition abilities of the laboratory-made and commercially available columns was discussed. PMID- 16359693 TI - Pore-network study of the characteristic periods in the drying of porous materials. AB - We study the periods that develop in the drying of capillary porous media, particularly the constant rate (CRP) and the falling rate (FRP) periods. Drying is simulated with a 3-D pore-network model that accounts for the effect of capillarity and buoyancy at the liquid-gas interface and for diffusion through the porous material and through a boundary layer over the external surface of the material. We focus on the stabilizing or destabilizing effects of gravity on the shape of the drying curve and the relative extent of the various drying periods. The extents of CRP and FRP are directly associated with various transition points of the percolation theory, such as the breakthrough point and the main liquid cluster disconnection point. Our study demonstrates that when an external diffusive layer is present, the constant rate period is longer. PMID- 16359694 TI - Convection, diffusion and reaction in a surface-based biosensor: modeling of cooperativity and binding site competition on the surface and in the hydrogel. AB - We study theoretically the transport and kinetic processes underlying the operation of a biosensor (particularly the surface plasmon sensor "Biacore") used to study the surface binding kinetics of biomolecules in solution to immobilized receptors. Unlike previous studies, we concentrate mainly on the modeling of system-specific phenomena rather than on the influence of mass transport limitations on the intrinsic kinetic rate constants determined from binding data. In the first problem, the case of two-site binding where each receptor unit on the surface can accommodate two analyte molecules on two different sites is considered. One analyte molecule always binds first to a specific site. Subsequently, the second analyte molecule can bind to the adjacent unoccupied site. In the second problem, two different analytes compete for one binding site on the same surface receptor. Finally, the third problem considers the case of positive cooperativity among bound molecules in the hydrogel using a simple mean field approach. The transport in both the flow channel and the hydrogel phases of the biosensor is taken into account in this case (with few exceptions, most previous studies assume a simpler model in which the hydrogel is treated as a planar surface with the receptors). We consider simultaneously diffusion and convection through the flow channel together with diffusion and cooperativity binding on the surface and in the hydrogel. In each case, typical results for the concentration contours of the free and bound molecules in the flow channel and hydrogel regions are presented together with the time-dependent association/dissociation curves and reaction rates. For binding site competition, the analysis predicts overshoot phenomena. PMID- 16359695 TI - Distribution of hexavalent Cr species across the clay mineral surface-water interface. AB - The adsorption isotherms of Cr(VI) on kaolinite, montmorillonite, and alumina were adequately treated with Langmuir model showing behavior characteristic of single-layer adsorption. The efficiency of the adsorbents in removing Cr(VI) from water follows the order alumina > kaolinite > montmorillonite > silica. Speciation studies indicate that hydrogen chromate ions were the major adsorbed species and simultaneous adsorption of dichromate ion occurred at concentrations greater than approximately 10(-3) mol L(-1). It is most probable that the mechanism of adsorption of the hydrogen chromate ion at the surface of alumina is predominantly electrostatic adsorption, with outer sphere complex formation. PMID- 16359696 TI - Inorganic particulates in removal of heavy metal toxic ions--part X: rapid and efficient removal of Hg(II) ions from aqueous solutions by hydrous ferric and hydrous tungsten oxides. AB - A radiotracer technique has been used to study the removal of Hg(II) ions from aqueous solutions by synthesized and well-characterized hydrous ferric oxide (HFO) and hydrous tungsten oxide (HTO). Adsorptive concentration (10(-4)-10(-8) mol dm(-3)), pH (ca 4.0-10.0) and temperatures (303-333 K) were examined for assessing optimal conditions for removal of these ions. The uptake of Hg(II) ions, fitted well with the Freundlich isotherm, increased with increasing temperature. No significant desorption took place in the present temperature range. The presence of other anions and cations affected the uptake of Hg(II) markedly. Irradiation of hydrous ferric oxide and tungsten oxide by using a 11.1 x 10(9)Bq (Ra-Be) neutron source having a neutron flux of 3.9 x 10(6)cm(-2)s(-1) with the associated gamma-dose rate of 1.72 Gy h(-1) did not influence the extent of adsorption of Hg(II) significantly. PMID- 16359697 TI - Preparation and characterization of a new layered double hydroxide, Co-Zr-Si. AB - The layered double hydroxides (LDHs) are nano-ordered layered compounds and well known for their ability to intercalate anionic compounds. Most LDH is prepared conventionally only with divalent and trivalent cations. In this study, Co-Zr-Si LDH, consisting of divalent, tetravalent, and tetravalent cations, was prepared and reacted with monocarboxylic acids at room temperature. The Co-Zr-Si LDH and intercalated compounds have been characterized by energy-dispersive X-ray spectrometry, X-ray powder diffraction, IR spectra, thermal analysis, and scanning electron microscopy (SEM). The insertion of cyanate and carbonate anions into LDH was confirmed by IR spectra. XRD patterns of the prepared Co-Zr-Si LDH showed that the interlayer spacing of the LDH is 0.78 nm. The spacing is similar to that of usual LDH in which chloride, carbonate, or bromide anion is the guest. SEM images showed that Co-Zr-Si LDH can exist as plate-like or fibrous structures. PMID- 16359698 TI - Effect of host size on male fitness in the parasitoid wasp Dinarmus basalis. AB - The effect of host size on male fitness was tested in the parasitoid wasp Dinarmus basalis (Hymenoptera, Pteromalidae) using hosts of different fresh weight. Fitness was measured as the sperm stock in seminal vesicles, and the ability to access females in single or competition situations. Both body size and sperm in seminal vesicles increased with host fresh weight. Males from small hosts had a reduced size and sperm stock compared to those from larger hosts. In single situations, males from both small and large hosts had similar reproductive capacities, whereas in multiple mating or competition situations, males from small hosts were at a disadvantage, inseminating fewer females and copulating less frequently. However, females did not appear to choose between males, and no effect on sperm stored in the spermatheca was observed. Being small does not prevent a D. basalis male mating and producing progeny in single situations, although more offspring could be expected from larger males because of their better competitive abilities. PMID- 16359699 TI - Role of membrane transport of water and glycerol in the freeze tolerance of the rice stem borer, Chilo suppressalis Walker (Lepidoptera: Pyralidae). AB - Overwintering larvae of the rice stem borer, Chilo suppressalis accumulate glycerol and are freezing tolerant to about -25 degrees C. However, non diapausing larvae cannot accumulate glycerol and are killed by freezing. We compared the extent of tissue damage, the effects of glycerol concentration, and the transport of glycerol and water in fat body tissues from these larvae at selected freezing temperatures. Tissues from overwintering larvae, but not non diapausing larvae, survive when frozen at -20 degrees C with 0.25 M glycerol, but the protection afforded by glycerol is offset by the water-channel inhibitor mercuric chloride. Glycerol in higher concentration (0.75 M) affords some protection even to the fat body of non-diapausing larvae. Radiotracer assays of overwintering larvae show that water leaves the tissues during freezing while glycerol enters, and that mercuric chloride disrupts this process. Transport is also disrupted after lethal freezing at -35 degrees C. Therefore, membrane transport of water and glycerol is involved in the avoidance of freezing injury to fat body cells of the rice stem borer, apparently by mediating the replacement of water with glycerol in freezing-tolerant tissues. PMID- 16359700 TI - Structural determinants of rotavirus subgroup specificity mapped by cryo-electron microscopy. AB - The rotavirus double-layered particle (DLP) is a molecular machine that transcribes 11 genomic segments of double-stranded RNA into full-length mRNA segments during viral replication. DLPs from the human Wa strain of virus, belonging to subgroup II (SG II), possess a significantly reduced level of transcriptase activity compared to bovine UK DLPs that belong to subgroup I (SG I). Cryo-electron microscopy and icosahedral image analysis was used to define the structural basis for this difference in transcriptase activity and to derive three-dimensional density maps of bovine UK and human Wa DLPs at 26 angstroms and 28 angstroms resolution, respectively. The two rotavirus strains had the same diameter, T = 13 l icosahedral lattice symmetry and size of the VP6 trimers on the surface of the DLPs. However, the Wa particles displayed a remarkable absence of VP6 trimers surrounding each 5-fold vertex position. To further explore these structural differences, three-dimensional reconstructions were generated of DLPs decorated with Fab fragments derived from subgroup-specific monoclonal antibodies. The X-ray structures of VP6 and a generic Fab fragment were then docked into the cryo-electron microscopy density maps, which allowed us to propose at "pseudo-atomic" resolution the locations of the amino acid residues defining the subgroup-specific epitopes. Our results demonstrate a correlation between the structure of the VP6 layer and the transcriptase activity of the particles, and suggest that the stability of VP6 trimers, specifically those at the icosahedral 5-fold axes, may be critical for mRNA synthesis. Thus, subgroup specificity of rotavirus may reflect differences in the architecture of the double-layered particle, with resultant consequences for viral mRNA synthesis. PMID- 16359701 TI - Crystal structure of the S.cerevisiae exocyst component Exo70p. AB - The exocyst is an evolutionarily conserved multiprotein complex required for the targeting and docking of post-Golgi vesicles to the plasma membrane. Through its interactions with a variety of proteins, including small GTPases, the exocyst is thought to integrate signals from the cell and signal that vesicles arriving at the plasma membrane are ready for fusion. Here we describe the three-dimensional crystal structure of one of the components of the exocyst, Exo70p, from Saccharomyces cerevisiae at 3.5A resolution. Exo70p binds the small GTPase Rho3p in a GTP-dependent manner with an equilibrium dissociation constant of approximately 70 microM. Exo70p is an extended rod approximately 155 angstroms in length composed principally of alpha helices, and is a novel fold. The structure provides a first view of the Exo70 protein family and provides a framework to study the molecular function of this exocyst component. PMID- 16359702 TI - Crystal structure of human guanosine monophosphate reductase 2 (GMPR2) in complex with GMP. AB - Guanosine monophosphate reductase (GMPR) catalyzes the irreversible and NADPH dependent reductive deamination of GMP to IMP, and plays a critical role in re utilization of free intracellular bases and purine nucleosides. Here, we report the first crystal structure of human GMP reductase 2 (hGMPR2) in complex with GMP at 3.0 A resolution. The protein forms a tetramer composed of subunits adopting the ubiquitous (alpha/beta)8 barrel fold. Interestingly, the substrate GMP is bound to hGMPR2 through interactions with Met269, Ser270, Arg286, Ser288, and Gly290; this makes the conformation of the adjacent flexible binding region (residues 268-289) fixed, much like a door on a hinge. Structure comparison and sequence alignment analyses show that the conformation of the active site loop (residues 179-187) is similar to those of hGMPR1 and inosine monophosphate dehydrogenases (IMPDHs). We propose that Cys186 is the potential active site, and that the conformation of the loop (residues 129-133) suggests a preference for the coenzyme NADPH over NADH. This structure provides important information towards understanding the functions of members of the GMPR family. PMID- 16359703 TI - Changing the specificity of a bacterial chemoreceptor. AB - The methyl-accepting chemotaxis proteins are a family of receptors in bacteria that mediate chemotaxis to diverse signals. To explore the plasticity of these proteins, we have developed a simple method for selecting cells that swim to target attractants. The procedure is based on establishing a diffusive gradient in semi-soft agar plates and does not require that the attractant be metabolized or degraded. We have applied this method to select for variants of the Escherichia coli aspartate receptor, Tar, that have a new or improved response to different amino acids. We found that Tar can be readily mutated to respond to new chemical signals. However, the overall change in specificity depended on the target compound. A Tar variant that could detect cysteic acid still showed a strong sensitivity to aspartate, indicating that the new receptor had a broadened specificity relative to wild-type Tar. Tar variants that responded to phenylalanine or N-methyl aspartate, or that had an increased sensitivity to glutamate showed a strong decrease in their response to aspartate. In at least some of the cases, the maximal level of sensitivity that was obtained could not be attributed solely to substitutions within the binding pocket. The new tar alleles and the techniques described here provide a new approach for exploring the relationship between ligand binding and signal transduction by chemoreceptors and for engineering new receptors for applications in biotechnology. PMID- 16359704 TI - Structure-based prediction of bZIP partnering specificity. AB - Predicting protein interaction specificity from sequence is an important goal in computational biology. We present a model for predicting the interaction preferences of coiled-coil peptides derived from bZIP transcription factors that performs very well when tested against experimental protein microarray data. We used only sequence information to build atomic-resolution structures for 1711 dimeric complexes, and evaluated these with a variety of functions based on physics, learned empirical weights or experimental coupling energies. A purely physical model, similar to those used for protein design studies, gave reasonable performance. The results were improved significantly when helix propensities were used in place of a structurally explicit model to represent the unfolded reference state. Further improvement resulted upon accounting for residue-residue interactions in competing states in a generic way. Purely physical structure based methods had difficulty capturing core interactions accurately, especially those involving polar residues such as asparagine. When these terms were replaced with weights from a machine-learning approach, the resulting model was able to correctly order the stabilities of over 6000 pairs of complexes with greater than 90% accuracy. The final model is physically interpretable, and suggests specific pairs of residues that are important for bZIP interaction specificity. Our results illustrate the power and potential of structural modeling as a method for predicting protein interactions and highlight obstacles that must be overcome to reach quantitative accuracy using a de novo approach. Our method shows unprecedented performance in predicting protein-protein interaction specificity accurately using structural modeling and suggests that predicting coiled-coil interactions generally may be within reach. PMID- 16359705 TI - Fold independent structural comparisons of protein-ligand binding sites for exploring functional relationships. AB - The rapid growth in protein structural data and the emergence of structural genomics projects have increased the need for automatic structure analysis and tools for function prediction. Small molecule recognition is critical to the function of many proteins; therefore, determination of ligand binding site similarity is important for understanding ligand interactions and may allow their functional classification. Here, we present a binding sites database (SitesBase) that given a known protein-ligand binding site allows rapid retrieval of other binding sites with similar structure independent of overall sequence or fold similarity. However, each match is also annotated with sequence similarity and fold information to aid interpretation of structure and functional similarity. Similarity in ligand binding sites can indicate common binding modes and recognition of similar molecules, allowing potential inference of function for an uncharacterised protein or providing additional evidence of common function where sequence or fold similarity is already known. Alternatively, the resource can provide valuable information for detailed studies of molecular recognition including structure-based ligand design and in understanding ligand cross reactivity. Here, we show examples of atomic similarity between superfamily or more distant fold relatives as well as between seemingly unrelated proteins. Assignment of unclassified proteins to structural superfamiles is also undertaken and in most cases substantiates assignments made using sequence similarity. Correct assignment is also possible where sequence similarity fails to find significant matches, illustrating the potential use of binding site comparisons for newly determined proteins. PMID- 16359706 TI - The MS2 coat protein shell is likely assembled under tension: a novel role for the MS2 bacteriophage A protein as revealed by small-angle neutron scattering. AB - Recombinant forms of the bacteriophage MS2 and its RNA-free (empty) MS2 capsid were analyzed in solution to determine if RNA content and/or the A (or maturation) protein play a role in the global arrangement of the virus protein shell. Analysis of the (coat) protein shell of recombinant versions of MS2 that lack the A protein revealed dramatic differences compared to wild-type MS2 in solution. Specifically, A protein-deficient virus particles form a protein shell of between 31(+/-1) A and 37(+/-1) A. This is considerably thicker than the protein shell formed by either the wild-type MS2 or the RNA-free MS2 capsid, whose protein shells have a thickness of 21(+/-1) A and 25(+/-1) A, respectively. Since the A protein is known to separate from the intact MS2 protein shell after infection, the thin shell form of MS2 represents the pre-infection state, while the post-infection state is thick. Interestingly, these A protein-dependent differences in the virus protein shell are not seen using crystallography, as the crystallization process seems to artificially compact the wild-type MS2 virion. Furthermore, when the A protein is absent from the virus shell (post-infection), the process of crystallization exerts sufficient force to convert the protein shell from the post-infection (thick) state to the pre-infection (thin) conformation. In summary, the data are consistent with the idea that RNA content or amount does not affect the structure of the MS2 virus shell. Rather, the A protein influences the global arrangement of the virus coat dramatically, possibly by mediating the storage of energy or tension within the protein shell during virus assembly. This tension may later be used to eject the MS2 genomic RNA and A protein fragments into the host during infection. PMID- 16359707 TI - How evolutionary pressure against protein aggregation shaped chaperone specificity. AB - As protein aggregation is potentially lethal, control of protein conformation by molecular chaperones is essential for cellular organisms. This is especially important during protein expression and translocation, since proteins are then unfolded and therefore most susceptible to form non-native interactions. Using TANGO, a statistical mechanics algorithm to predict protein aggregation, we here analyse the aggregation propensities of 28 complete proteomes. Our results show that between 10% and 20% of the residues in these proteomes are within aggregating protein segments and that this represents a lower limit for the aggregation tendency of globular proteins. Further, we show that not only evolution strongly pressurizes aggregation downwards by minimizing the amount of strongly aggregating sequences but also by selectively capping strongly aggregating hydrophobic protein sequences with arginine, lysine and proline. These residues are strongly favoured at these positions as they function as gatekeepers that are most efficient at opposing aggregation. Finally, we demonstrate that the substrate specificity of different unrelated chaperone families is geared by these gatekeepers. Chaperones face the difficulty of having to combine substrate affinity for a broad range of hydrophobic sequences and selectivity for those hydrophobic sequences that aggregate most strongly. We show that chaperones achieve these requirements by preferentially binding hydrophobic sequences that are capped by positively charged gatekeeper residues. In other words, targeting evolutionarily selected gatekeepers allows chaperones to prioritize substrate recognition according to aggregation propensity. PMID- 16359709 TI - A pathway for the transmission of allosteric signals in the ribosome through a network of RNA tertiary interactions. AB - There are a large number of tertiary contacts between nucleotides in 23S rRNA, but which are of functional importance is not known. Disruption of one between A2662 in the sarcin/ricin loop (SRL) and A2531 in the peptidyl-transferase center (PTC) has adverse effects on cell growth and on the ability of ribosomes to catalyze some but not other partial reactions of elongation. A lethal A2662C mutation is suppressed by a concomitant lethal A2531 mutation. Ribosomes with non lethal A2531 mutations, treated with base-specific reagents, have alterations of nucleotides in the PTC (home of A2531) and, more significantly, in nucleotides in the SRL and in the GTPase center. The results suggest that the function of ribosomal centers is coordinated by a set of sequential conformational changes in rRNA that are a response to signals transmitted through a network of tertiary interactions. PMID- 16359708 TI - Functional analysis of the post-transcriptional regulator RsmA reveals a novel RNA-binding site. AB - The RsmA family of RNA-binding proteins are global post-transcriptional regulators that mediate extensive changes in gene expression in bacteria. They bind to, and affect the translation rate of target mRNAs, a function that is further modulated by one or more, small, untranslated competitive regulatory RNAs. To gain new insights into the nature of this protein/RNA interaction, we used X-ray crystallography to solve the structure of the Yersinia enterocolitica RsmA homologue. RsmA consists of a dimeric beta barrel from which two alpha helices are projected. From structure-based alignments of the RsmA protein family from diverse bacteria, we identified key amino acid residues likely to be involved in RNA-binding. Site-specific mutagenesis revealed that arginine at position 44, located at the N terminus of the alpha helix is essential for biological activity in vivo and RNA-binding in vitro. Mutation of this site affects swarming motility, exoenzyme and secondary metabolite production in the human pathogen Pseudomonas aeruginosa, carbon metabolism in Escherichia coli, and hydrogen cyanide production in the plant beneficial strain Pseudomonas fluorescens CHA0. R44A mutants are also unable to interact with the small untranslated RNA, RsmZ. Thus, although possessing a motif similar to the KH domain of some eukaryotic RNA-binding proteins, RsmA differs substantially and incorporates a novel class of RNA-binding site. PMID- 16359710 TI - Control of attention in schizophrenia. AB - The deficits of attention contribute significantly to the clinical picture of functional disability seen in schizophrenia, but there is no consensus as to whether this cognitive function can be fractionated to allow further characterisation of the impairment. We examined fifteen patients with chronic schizophrenia and fifteen controls using paired tasks designed to measure four hypothetical aspects of attentional control: the ability to focus attention, to resist distraction, to shift attention, and to divide attention. The group with schizophrenia showed a significant improvement in accuracy on a digit span repetition task when a simultaneous box-crossing task was added (divided attention condition). Although the patient group showed impaired performance across nearly all of the tasks, they were not disproportionately impaired during the task conditions assumed to demand greater attention. These results suggest that the aspects of attention and executive function under study are not significantly affected by schizophrenia and indicate the need for further characterisation of the impairment usually reported using conventional tests of attention on those with schizophrenia. PMID- 16359711 TI - Utilizing gamma isotope tracers to determine sediment source at reef sites near the Charleston Ocean Dredged Material Disposal Site. AB - The Charleston, South Carolina Ocean Dredged Material Disposal Site (ODMDS) has been heavily utilized as a disposal site for dredged material resulting from maintenance and channel deepening in the Charleston Harbor. Continuous monitoring by the South Carolina Department of Natural Resources at the ODMDS has indicated the presence of fine-grained sediment within the monitoring zones. However, since the Charleston Harbor is formed by the conjunction of three rivers, it has been suggested that some of the fine-grained sediment surrounding the ODMDS could be due to river transport rather than solely by disposal activities. In order to trace the outflow of sediment from the harbor, natural and man-made isotopes were utilized. (7)Be (natural cosmogenic isotope) and (137)Cs (man-made isotope) are often associated with estuarine sediments. Both isotopes were used as tracers in an attempt to determine the extent of density driven sediment flow from the Charleston Harbor. (7)Be was detected in many of the offshore sampling stations indicating a direct correlation to the harbor. (137)Cs was only found in one sediment trap sample offshore, but none the less indicated some transport from the harbor. Further study for utilizing isotopic tracers in determining offshore sediment transport is still being conducted at the disposal site. It is anticipated that further (7)Be and (137)Cs isotopic monitoring offshore Charleston will aid in determining the role that tidal and density driven sediments play in the sediment budgets at the hard bottom reef sites. PMID- 16359712 TI - Occurrence and stress response of N-methylproline compounds in Tamarix species. AB - A number of N-methylproline analogues have been found to accumulate in different species of Tamarix. These include N-methyl-L-proline (MP), trans-4-hydroxy-N methyl-L-proline (M4HP) and trans-3-hydroxy-N-methyl-L-proline (M3HP). The three compounds appeared in all species but their relative and absolute levels depend upon species, ecotype and level of applied salt stress. A salt-conditioned ecotype of T. jordanis (Sodom) dramatically increased its accumulation of all proline analogues when subject to salt stress whereas a non-saline ecotype (Gilboa) showed little effect. The levels of M4HP and M3HP in T. meyeri increased with increasing salt stress whereas MP levels remained almost constant. PMID- 16359713 TI - Scaling up of PCR-based surveillance of bacterial meningitis in the African meningitis belt: indisputable benefits of multiplex PCR assay in Niger. AB - The absence of reliable laboratories for culture of Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae, the three main causes of bacterial meningitis in Africa, hampers microbiological surveillance in these countries. To compensate for this situation in Niger, a multiplex single-tube PCR method has been implemented at a central level to test cerebrospinal fluid (CSF) samples. The overall confirmation rate for PCR (N=3791) was 40.8% compared with 16.0% for culture (N=945) (P<10(-6)). Among 850 CSF specimens tested by both methods, the overall confirmation rate was 29.4% for PCR and 16.4% for culture (P<10(-8)). PCR was also efficient for the CSF specimens stored in Trans-isolate medium. In conclusion, PCR assay is currently a key tool in Africa to improve microbiological surveillance of bacterial meningitis. PMID- 16359714 TI - Applied and basic research on the epidemiology, morbidity, and immunology of schistosomiasis in fishing communities on Lake Albert, Uganda. AB - We report multidisciplinary studies on schistosomiasis which have been ongoing in the fishing communities of Piida, Booma, Bugoigo and Walakuba, on Lake Albert, Uganda, since 1996. Schistosomiasis is the major health problem in this area, with high infection intensities and prevalence. In addition to generating basic data on the epidemiology, morbidity and immunology of human schistosomiasis, this research programme is providing important descriptive and methodological information, and has contributed to the increase in operational capacity within Uganda in recent years. Such information and operational capacity are needed to facilitate much needed schistosomiasis control programmes, such as the Schistosomiasis Control Initiative that was launched in Uganda in 2003. PMID- 16359715 TI - A direct antigen heterologous enzyme immunoassay for measuring progesterone in serum without using displacer. AB - An antigen heterologous enzyme-linked immunosorbent assay (ELISA) for directly measuring progesterone in serum is described. Six combinations of antigens and enzyme conjugates were tested; the enzyme conjugate 17-alphaOH-progesterone-3-O carboxymethyloxime-alkalinephosphatase (17-alphaOH-P-3-CMO-ALP) and the immunogen progesterone-3-carboxymethyloxime-bovine serum albumin (P-3-CMO-BSA) were found to be best. Fifty microliters of standard or serum sample and 100 microL of the 17-alphaOH-P-3-CMO-ALP enzyme conjugate were added to the antibody coated wells, and incubated for 1 h at 37 degrees C. Bound enzyme activity was measured by using p-nitrophenyl phosphate as substrate. The sensitivity of the assay was 0.11 ng/mL, and intra- and inter-assay CVs ranged from 5.1% to 9.6%. The analytical recoveries were 97-105%. The serum progesterone values obtained by this method correlated well with those obtained by radioimmunoassay; r=0.97 (n=44). Moreover, in this ELISA no displacing agent was used or special means was required to displace progesterone from corticosteroid binding globulin (CBG). Serum progesterone concentrations of subjects, with histories of recurrent spontaneous abortions were also measured, and correlated well with clinical history. PMID- 16359716 TI - Cloning and expression of a novel cDNA encoding a mannose-specific jacalin related lectin from Oryza sativa. AB - Lectin plays an important role in defense signaling in plants. A few genes of this family have been cloned. Here we report on a mannose-specific jacalin related lectin in rice. Using sequence information of wheat gene VER2, which we had previously cloned, we were able to amplify a cDNA of OsJAC1 from Oryza sativa by RT-PCR. The cDNA of OsJAC1 was 1172 bp and contained a 921-bp open reading frame (ORF) encoding dirigent (amino acids 26-139) and jacalin (amino acids 175 305) domains of 306 amino acids. Comparison of the OsJAC1 sequence with those of other lectins (jacalin) from rice, wheat and other species revealed that OsJAC1 had the 12 amino acid positions conserved in all mannose-binding lectins. Semi quantitative RT-PCR revealed that OsJAC1 expression was present in stems, leaves and young spikes but not young roots; the expression was high in leaves and low in stems and spikes. And methyl jasmonate could induce the expression of OsJAC1. To test the activity of OsJAC1, the jacalin domain at the C-terminal was expressed in E. coli. BL21 (DE3). The purified recombinant protein could agglutinate red blood cells of rabbit, and the agglutination activity was strongly inhibited by mannose compared with other carbohydrates. These results indicate that lectin with dirigent and jacalin domains exists in rice as well as wheat. This is the first report of cDNA cloning of mannose-binding jacalin related lectin with a dirigent domain in N-terminal region from O. sativa. PMID- 16359717 TI - Neutralisation of venom-induced haemorrhage by IgG from camels and llamas immunised with viper venom and also by endogenous, non-IgG components in camelid sera. AB - Envenoming by snakes results in severe systemic and local pathology. Intravenous administration of antivenom, prepared from IgG of venom immunised horses or sheep, is the only effective treatment of systemic envenoming. Conventional antivenoms, formulated as intact IgG, papain-cleaved (Fab) or pepsin-cleaved F(ab')2 fragments, are however ineffective against the local venom effects because of their inability to penetrate the blood/tissue barrier. We have embarked on a new research program to examine (i) whether the unusually small (15 kDa) antigen-binding fragment of camelid heavy chain IgG (V(H)H) can be exploited to neutralise the local effects of envenoming and (ii) whether a novel antivenom to treat both the systemic and local effects of envenoming can be formulated by combining anti-snake venom V(H)H and conventional F(ab')2. In this preliminary study, we demonstrate that camels and llamas respond to immunisation with Echis ocellatus venom with high antibody titres and broad antigen specificity. These encouraging immunological results were matched by the successful elimination of venom-induced haemorrhage by IgG from the venom-immunised camels and llamas. Unexpectedly, we report for the first time that camelid serum contains a non-IgG, highly potent inhibitor of venom-induced haemorrhage. PMID- 16359718 TI - Purification and properties of hyaluronidase from Palamneus gravimanus (Indian black scorpion) venom. AB - Scorpion venoms are a rich source of enzymes. Some of the enzymes such as phospholipase A2, proteolytic enzymes and phosphodiesterase are well characterized. However, hyaluronidase has not been studied extensively. In this paper we describe the purification and characterization of hyaluronidase (Hyaluronate lyase, E.C.3.2.1.35) from the Palamneus gravimanus scorpion venom by a combination of gel filtration on Sephadex G-75 and ion-exchange chromatography on DEAE-cellulose. The optimal pH and temperature for its maximum activity of the isolated enzyme were 4.5 and 37 degrees C, respectively, and its K(m) was 47.61 microg/ml at 37 degrees C and its specific activity was 6411.7 +/- 117TRU/min per mg against 250 +/- 4.0 TRU/min per mg for the whole desiccated venom suggesting 25-fold purification. The molecular weight of the isolated enzyme was 52 +/- 1 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography on Sephadex G-75. The enzyme was stable for 30 days in the presence of NaCl; no loss of activity was observed up to 37 degrees degrees C and showed a sharp decrease in its activity at 40 degrees C. Heparin inhibited the enzyme activity. PMID- 16359719 TI - The effect of experimental Bothrops jararaca envenomation on pregnant mice. AB - The injury caused by the intramuscular injection of a single dose of Bothrops jararaca venom (0.24 mg/kg body weight) to mice on day 8 of pregnancy and examined on day 9 was investigated. Macroscopic and histological examination showed that the bothropic venom caused an increase in the incidence of fetal resorptions. Histologically, a characteristic involution of mature decidua was noticed in saline-treated mice; however, necrotic trophoblast giant cells and decidual cells were also present in this region of mice treated with B. jararaca venom, mainly close to the embryo. Hemorrhagic areas were also observed at maternal-fetal interface, which contained maternal erythrocytes and polymorphonuclears. Plasma fibrinogen levels were lower in envenomed group (p < or = 0.0001), but prothrombin time and activated partial thromboplastin time remained unaltered. Total and differential white blood cell counts were not statistically different between groups. Thus, B. jararaca venom causes injuries not only to the fetus, but also to decidual tissue and blood coagulation of pregnant mice. It is not clear, nonetheless, whether disturbances during the development of pregnancy are due to a direct effect of venom on uterus/fetus or to homeostatic changes in dams, such as clotting disturbances, or to both of them. PMID- 16359721 TI - Second-order motion shifts perceived position. AB - Many studies have documented that first-order motion influences perceived position. Here, we show that second-order (contrast defined) motion influences the perceived positions of stationary objects as well. We used a Gabor pattern as our second-order stimulus, which consisted of a drifting sinusoidal contrast modulation of a dynamic random-dot background; this second-order carrier was enveloped by a static Gaussian contrast modulation. Two vertically aligned Gabors had carrier motion in opposite directions. Subjects judged the relative positions of the Gabors' static envelopes. The positions of the Gabors appeared shifted in the direction of the carrier motion, but the effect was narrowly tuned to low temporal frequencies across all tested spatial frequencies. In contrast, first order (luminance defined) motion shifted perceived positions across a wide range of temporal frequencies, and this differential tuning could not be explained by differences in the visibility of the patterns. The results show that second-order motion detection mechanisms contribute to perceived position. Further, the differential spatial and temporal tuning of the illusion supports the idea that there are distinct position assignment mechanisms for first and second-order motion. PMID- 16359720 TI - Photometric determinants of perceived transparency. AB - Photometric constraints for the perception of transparency were investigated using stereoscopic textured displays. A contrast discontinuity divided the textured displays into two lateral halves, with one (reference) half fixed. Observers adjusted the luminance range within the other (test) half in order to perform two tasks: (i) indicate the highest luminance range for which the test side is perceived to be transparent, and (ii) indicate the lowest luminance range for which the test side is seen as being in plain view. Settings were obtained for multiple values of test mean luminance, in order to map out the perceptual locus of transition between transparency and non-transparency. The results revealed a systematic violation of Metelli's magnitude constraint in predicting the percept of transparency. Observer settings were approximated instead by a constraint based on perceived contrast (which matched Michelson contrast for the textures used). The results also revealed large asymmetries between darkening and lightening transparency. When the test was darker than the reference, settings were highly consistent across observers and closely followed the Michelson contrast prediction. When the test was lighter, however, there was greater variability across observers, with two observers exhibiting shifts toward Metelli's magnitude constraint. Moreover, each observer's setting reliability was significantly worse for lightening transparency than darkening transparency. These results suggest that (polarity-preserving) darkening serves as an additional cue to perceptual transparency. PMID- 16359722 TI - Prenatal ethanol exposure: sex differences in anxiety and anxiolytic response to a 5-HT1A agonist. AB - This study utilized a novelty-induced suppression of feeding task to examine anxiety-like behaviour and the anxiolytic effects of the 5-HT1A agonist, 8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), in rats prenatally exposed to ethanol. Adult offspring from ethanol exposed (E), pair-fed control (PF) and ad libitum-fed control (C) dams were habituated to a novel palatable food for 21 days and measures of baseline feeding obtained. On day 22 (d 22), animals received either 8-OH-DPAT (0.06 mg/kg) or vehicle (0.9% NaCl) and feeding behaviour in the home cage or a novel cage was observed. Factor analyses revealed that feeding behaviour on d 21 (habituation) and d 22 (test day) are reflective of two different affective states, and that the single factor that emerged for novel cage testing on d 22 likely reflects the anxiety evoked by the novel test condition. Analyses of variance on the variables loading significantly onto the factors support the suggestion that the novel environment is anxiogenic for both females and males, and that 8-OH-DPAT acts as an anxiolytic. However, although both females and males showed alterations in behaviours (latency, amount, duration of feeding) reflective of anxiety, 8-OH-DPAT had anxiolytic effects primarily in females. Importantly, prenatal ethanol exposure altered several aspects of behavior in this task. Both E females and males consumed less than their control counterparts on d 21, suggesting a possible delay or deficit in response habituation. During home cage testing on d 22, overall feeding rate was slower in E than in C females, and E males consumed less than PF and C males. In addition, a smaller percentage of E than PF and C females fed in the novel environment, and latency to feed was significantly increased in E compared to control females. These findings indicate that prenatal ethanol exposure results in increased anxiety-like behaviour in adulthood, and that prenatal ethanol induced hyponeophagia may be, at least in part, mediated by the 5-HT1A receptor. This study is one of the first to demonstrate specific increases in anxiety-like behaviour in animals prenatally exposed to ethanol, and further supports the utility of the novelty-induced suppression of feeding task in assessing anxiety and the effectiveness of anxiolytic agents. PMID- 16359723 TI - Separation-induced ultrasonic vocalization in rat pups: further pharmacological characterization. AB - In rat pups, ultrasonic vocalizations were emitted in response to separation from the mothers, littermates, and nest. It has been suggested that these separation induced ultrasonic vocalizations (SIV) in rat pups form one of the animal models of anxiety. The primary aim of the present study is to investigate the effect of the compounds acting on stress-related peptide receptors such as a vasopressin V1b receptor antagonist and a corticotropin-releasing factor CRF1 receptor antagonist in rat pup SIV. The secondary objective is to establish further confirmation of the predictive validity of SIV testing. Both the V1b receptor antagonist SSR149415 and the CRF1 receptor antagonist CP154,526 reduced SIV, suggesting antagonists for stress-related peptide receptors are effective in this model. Furthermore, as with selective serotonin reuptake inhibitors such as fluvoxamine and paroxetine, SIV was also reduced by the serotonin and noradrenaline reuptake inhibitor milnacipran and the metabotropic glutamate receptor 5 antagonist MPEP, while desipramine was without effect. Thus, the present experiment highlights the important role of the stress-related peptide systems as well as of the serotonergic systems in SIV. Moreover, the present data support the usefulness of SIV for evaluating the anxiolytic-like activity of mechanically diverse compounds. PMID- 16359724 TI - Baclofen decreases feeding in non-human primates. AB - This study examined how the GABA(b) agonist baclofen (0.5-5.6 mg/kg, p.o.), reported to increase food intake in rodents, affected the appetitive and consummatory aspects of feeding of non-human primates. Baboons had access to food 24 h each day, but they had to complete a two-phase operant procedure in order to eat. Responding on one lever during a 30-min appetitive phase was required before animals could start a consumption phase, where responding on another lever led to food delivery, i.e., a meal. Responding during the appetitive phase resulted in presentations of food-related stimuli only. Baclofen increased the latency to the first meal and decreased both appetitive and consummatory behavior. At the largest dose, baclofen induced emesis, indicating that the effects were due to malaise rather than a specific motivational action. In contrast, the positive control diazepam (GABA(a) agonist, 1.0-2.0 mg/kg, i.m.) decreased the latency to the first meal and increased both appetitive (P < 0.07) and consummatory behavior. Although the baclofen-induced decrease in appetitive behavior replicates data obtained in rodents, the baclofen-induced decreases in consummatory behavior do not. The findings suggest that the effects of large doses of baclofen in non-human primates may, in part, be due to non-specific behavioral disruptions. PMID- 16359725 TI - Effects of semen extender and semen processing on motility and viability of frozen-thawed dog spermatozoa. AB - The aims of the present study were, to assess the effects of semen centrifugation, two different diluents and two different freezing methods on post thaw semen quality in canine semen, and to elucidate the interdependence of these parameters. For this purpose, the sperm-rich fractions of ejaculates from 12 healthy male beagles were divided into four aliquots. Two aliquots were centrifuged and resuspended with two TRIS-egg yolk based extenders: with Uppsala and Gill extender (Gill). The diluents differed in the concentration of glycerol and in the admixture of Equex STM paste (Nova Chemical Sales Inc., Scituate, MA, USA). Diluted semen was frozen either in a styrofoam box or with a computerized freezing machine and an optimized freezing curve (IceCube 1,810; Sy-Lab, Purkersdorf, A). The change in temperature inside the straws was measured during the freezing procedure. Thawed semen samples were assessed for motility and viability (SYBR-14/PI) using the computer assisted sperm analyzer SpermVision (Minitub, G) and a modified triple staining technique (flow cytometry). Deep freezing in the machine resulted in better motility and viability than in the box. The combination centrifugation-Uppsala extender-machine was superior to all other combinations, which was most evident after storage at +5 degrees C for 7 h (motility: 53.1%, viability: 64.9%). Post-thaw longevity and progressive motility were significantly improved by the use of the here introduced freezing curve. This was shown to be partly caused by less pronounced fluctuations of temperature inside the straws when compared to box-freezing. PMID- 16359726 TI - Semantic weight and verb retrieval in aphasia. AB - Individuals with agrammatic aphasia may have difficulty with verb production in comparison to nouns. Additionally, they may have greater difficulty producing verbs that have fewer semantic components (i.e., are semantically "light") compared to verbs that have greater semantic weight. A connectionist verb production model proposed by Gordon and Dell (2003) learns through error correction to "divide the labor" between syntax and semantics. Verbs that are semantically heavier come to depend less on syntax and more on semantics. For lighter verbs, the reverse is true. We performed this study to clarify the role of semantic weight in aphasic verb production and to test the prediction from Gordon and Dell that a brain lesion that impairs the syntactic input to verb retrieval will impair lighter verbs more than heavier ones. Consistent with this prediction, we found that the decrement for lighter verbs was present in a group with agrammatic aphasia but not in a matched group without agrammatism. PMID- 16359727 TI - Sulphur partitioning in sediments and biodeposits below mussel rafts in the Ria de Arousa (Galicia, NW Spain). AB - Forty four core samples were analyzed to determine sediment particle size, total organic carbon (TOC), total inorganic carbon (TIC), total nitrogen and total sulphur. Sequential extraction of S was also carried out, differentiating AVS (acid volatile sulphide), elemental S, organic S and pyrite S. The results obtained show that the presence of mussel rafts causes intense changes in the physicochemical composition and properties of the sea floor in the Ria de Arousa. The percentage of silt and clay, TOC, TIC and total N were significantly higher in the biodeposit than in the sediment. In contrast, there were no differences between the biodeposit and the sediment in terms of pH (8.0-8.7) and redox potential. The sediment and biodeposit were always anoxic, with values of redox potentials below -100 mV. In accordance with these conditions, the dominant fraction of S was pyrite S (FeS(2)). The AVS fraction and elemental S were present at low concentrations, except in the uppermost part of each core. Pyrite was relatively stable when the biodeposit and sediment were maintained in suspension for 8 days in oxic sea water; unlike the AVS fraction, which disappeared within a few hours. PMID- 16359728 TI - Down modulation of TNF-alpha mRNA placental expression by AZT used for the prevention of HIV-1 mother-to-child transmission. AB - Mechanisms of HIV-1 in utero mother-to-child transmission (MTCT) protection provided by AZT are not completely understood. The placental cytokine network is involved in the control of HIV-1 in utero transmission but the effect of AZT on this network is unknown. To evaluate the effects of AZT on placental cytokine expression, the chorionic villi from HIV-1 uninfected women term placentae were cultured with 0, 100, and 2,000 ng/ml AZT. Tissue fragments were harvested at days 1, 4, and 7 to determine the level of cytokine mRNA by real-time RT-PCR. The viability and morphology of the placental histocultures were monitored by the expression of beta-human chorionic gonadotropin (beta-hCG) gene, lipopolysaccharide (LPS) activation, and microscopic examination. AZT at 2,000 ng/ml significantly down-regulated TNF-alpha mRNA expression at day 1 and day 4, but had no effect on beta-hCG, stromal cell-derived factor 1 (SDF-1), and IL-10 gene expression. AZT did not induce any deleterious impact on placental tissue structure. Furthermore, activation of chorionic villi by LPS for 24 h up regulated IL-10 and TNF-alpha mRNA expression. Down-regulation of TNF-alpha mRNA could represent a mechanism through which AZT can decrease the risk of HIV-1 MTCT, in addition to its direct effect on HIV-1 replication. PMID- 16359729 TI - Exercise, cognition and Alzheimer's disease: more is not necessarily better. AB - Regional hypoperfusion, associated with a reduction in cerebral metabolism, is a hallmark of Alzheimer's disease (AD) and contributes to cognitive decline. Cerebral perfusion and hence cognition can be enhanced by exercise. The present review describes first how the effects of exercise on cerebral perfusion in AD are mediated by nitric oxide (NO) and tissue-type plasminogen activator, the release of which is regulated by NO. A conclusion of clinical relevance is that exercise may not be beneficial for the cognitive functioning of all people with dementia if cardiovascular risk factors are present. The extent to which cardiovascular risk factors play a role in the selection of older people with dementia in clinical studies will be addressed in the second part of the review in which the effects of exercise on cognition are presented. Only eight relevant studies were found in the literature, emphasizing the paucity of studies in this field. Positive effects of exercise on cognition were reported in seven studies, including two that excluded and two that included patients with cardiovascular risk factors. These findings suggest that cardiovascular risk factors do not necessarily undo the beneficial effects of exercise on cognition in cognitively impaired people. Further research is called for, in view of the limitations of the clinical studies reviewed here. PMID- 16359730 TI - Molecular cloning and characterization of CD9 cDNA from cartilaginous fish, red stingray, Dasyatis akajei. AB - CD9 is a glycoprotein of the transmembrane 4 superfamily (TM4SF) and is involved in various cellular processes. In this study, we describe the isolation of the full-length cDNA encoding for CD9 molecule (daCD9) of red stingray, Dasyatis akajei. This 1252 bp cDNA was isolated from leukocyte cDNA library and contains 681 bp open reading frame encoding 226 amino acid residues. Amino acid sequences analysis and structure prediction display approximately 50% identity to higher vertebrates with the presence of conserved structures, including the four transmembrane domains and certain characteristic residues. Southern blot analysis shows that daCD9 exists as a single copy gene. Northern blot analysis reveals that daCD9 is highly expressed in gill and spleen although its expression can be found in other tissues suggesting daCD9 might play an important role in immune defense in this fish. PMID- 16359731 TI - Influence of the physiologically widespread substances on the oxidative activity of copper(II) complexes with sinefungin, a nucleoside antibiotic. AB - The oxidation-promoting reactivity of the Cu(II)-sinefungin complex in the presence of hydrogen peroxide was studied at pH 7.4, using N,N-dimethyl-p nitrosoaniline (NDMA), as well as plasmid DNA as target molecules. Mixture of the complex with H(2)O(2) was found to be an efficient oxidant, bleaching NDMA solution, and generating single- and double-strand breaks in DNA. The oxidative DNA damage was investigated also in the presence of varying amounts of glutathione, histidine, Gly-Gly-His peptide, H2A histone, and ascorbic acid, showing diverse influence of those substances on the cleavage extension. PMID- 16359732 TI - Regulatory mechanisms of cortical laminar development. AB - The developing forebrain exhibits a high degree of spatiotemporal regulation of proliferation and cell cycle exit in progenitor cells of its proliferative zones. This results in the balanced deployment of progenitors between asymmetric division, yielding postmitotic neurons and cycling progenitors, and terminal symmetric division, resulting in differentiated daughter cells. Radial glia have been demonstrated to be the principal neuronal progenitor of the cortical primordium. Lineage tracing studies employing real-time imaging in vivo have enhanced understanding of neuronal production and migration. Cortical projection neurons have been shown to arise from the radial migration of precursors generated in the dorsal telencephalon, whereas most interneurons derive from the germinal zone of the ventral telencephalon and migrate tangentially into the primordial cortex. Cells from both populations undergo diverse and complex sequences of migratory activity. Neuronal phenotypic potential is informed in progenitors prior to their last cell division. Laminar and regional fate potential of progenitors becomes progressively restricted with successive cell cycles. This process of neuronal fate specification is regulated by the interaction of programs of transcriptional regulation with extrinsic patterning signals according to time and region of the proliferative zone in which the final mitotic cycle occurs. PMID- 16359733 TI - The adrenal gland is a source of stress-induced circulating IL-18. AB - The present study compared plasma IL-18 levels between sham-operated and adrenalectomized mice following stress to investigate whether the adrenal gland contributes to the elevation of circulating IL-18 during stress. Two hours of stress provoked a robust, stressor-dependent, elevation of IL-18 mRNA and peptide in the adrenal gland in sham-operated mice. Consistently, levels of circulating mature IL-18 increased during stress and remained elevated for up to 6 h after stress. The stress-induced increase in circulating IL-18 was abolished by adrenalectomy. These findings demonstrate that the adrenal gland is required to achieve elevation of circulating IL-18 during stress. PMID- 16359734 TI - Diagnostic and management problems of laryngopharyngeal reflux disease in children. AB - OBJECTIVE: Reflux is a common pediatric disorder and an association between reflux and otolaryngological conditions has been described. However, to prove a causal relationship a pathophysiological pathway must be identified, diagnostic test with high specificity and sensitivity must be developed and conservative or surgical treatment of reflux should be shown to predictably improve the otolaryngological problems. This review study aims at examining the available evidence for the above controversial issues. METHODS: Articles on pediatric laryngopharyngeal reflux published in English during the last decade were searched using Ovid and PubMed. RESULTS: A lack of consensus was found in four separate but interdependent areas: clinical manifestations, diagnostic testing, interpretation of findings and treatment. Although clinical experience and uncontrolled case series suggest that laryngopharyngeal reflux may possibly contribute to apnea, recurrent upper respiratory infections, laryngeal symptoms (mainly laryngomalacia and subglottic stenosis), sinusitis and otitis convincing data are lacking. For pediatric studies, the diagnostic role of pH monitoring, barium esophagram, scintigraphy, impedance monitoring, laryngoscopic examination, laryngeal biopsy and symptom assessment questionnaires remain to be defined. Interpretation of pharyngeal reflux events is controversial and the lack of established normative values as well as the existing variability in the diagnostic criteria (reflux definition, duration and number of pathological reflux events) limits the ability to directly compare results. Proposed laryngopharyngeal reflux treatment (lifestyle modification, medical or surgical therapy) is mostly empiric, with no significant placebo-controlled trials of treatment and outcomes. CONCLUSIONS: Limited evidence exists to support a causative relationship between reflux and any otorhinolaryngological condition or the effectiveness of treatment. Epidemiological and large-scale prospective controlled studies are required to clarify these issues. PMID- 16359735 TI - The use of perioperative antibiotics in tonsillectomy: does it decrease morbidity? AB - OBJECTIVE: To assess the efficacy of perioperative antibiotics in decreasing post operative morbidity among patients undergoing tonsillectomy or adenotonsillectomy. DESIGN: Meta-analysis based on a structured search of the literature, using MEDLINE and the Cochrane database. SUBJECTS: Only articles, which included both treatment and control groups, were included in the final analysis. Studies were limited to ones that involved human subjects, including both children and adults. Studies using steroids and topical antibiotics were excluded. OUTCOMES: Articles were abstracted for patient factors, elements of study design, methods of patient assignment to treatment and control groups, and clinical outcomes. The primary outcome, time required for return to normal oral intake, was analyzed in the final meta-analysis. METHODS: Four hundred and twenty eight articles were initially identified. There were 23 potentially appropriate articles. Of these, 18 were able to be located in full text form and in English. Seven of these studies directly studied the efficacy of perioperative antibiotics (versus no antibiotics) in decreasing post-operative morbidity. Four studies had sufficient information to calculate effects estimates (xi) and standard deviations (Si) for the primary outcome. Three studies either did not report the outcome of interest or did not report a measure of stability (e.g. p-value or confidence interval). The data available from the first four studies were combined in a quantitative meta-analysis. Statistical analyses were performed using STATA for Windows software. RESULTS: The pooled estimate indicated that the antibiotic group returned to normal oral intake, on average, 1 day sooner than the controls. This difference was found to be statistically significant with a 95% confidence interval of 0.5-1.6 days. An additional assessment of three qualitative reports also suggested the use of perioperative antibiotics for adenotonsillectomy was associated with less post-operative pain. However, studies varied in terms of study quality, sample size, outcome examined, measure used and antibiotic administered. Definitive conclusions regarding the effect of perioperative antibiotics on other outcomes including bleeding, halitosis, fever, activity level and nausea and vomiting could not be drawn due to the small numbers of studies. CONCLUSIONS: In this meta-analysis, the use of perioperative antibiotics in patients who have had tonsillectomy or adenotonsillectomy appears to be associated with a 1-day reduction in the time required for return to normal oral intake. For other potentially important outcomes, such as post operative pain or bleeding, sufficient data were not available to make any definitive conclusions regarding the effect of perioperative antibiotics. PMID- 16359736 TI - CT and virtual endoscopy findings in congenital laryngeal web. AB - Laryngeal web is a rare lesion resulting from incomplete recanalization of the primitive larynx. Because the extent of airway involvement affects surgical management, patients should be studied thoroughly before treatment. Although the diagnosis is based on the endoscopy findings in most of the cases, laryngoscopy may fail to show the subglottic extension of the disease. Virtual endoscopy may provide the information needed for surgery in such cases. Here, we report the computed tomography and virtual endoscopy findings in a 5-year-old child with a laryngeal web. PMID- 16359737 TI - The effect of treatment with vincristine on transient evoked and distortion product otoacoustic emissions. AB - OBJECTIVE: Vincristine chemotherapy is mainly associated with neurotoxic effects. The ototoxicity of vincristine has been related to high dosage, while low and moderate doses do not seem to induce significant hearing impairment when measured by pure tone or speech audiometry. Otoacoustic emissions have been reported to be more sensitive in early detection of ototoxicity than conventional pure tone audiometry. The present study was directed at determining whether vincristine treatment interferes with outer hair cell function in the absence of measurable changes in pure tone audiometry. METHODS: We studied prospectively a cohort of ten children suffering from leukemia. All children were subjected to tympanogram, stapedial muscle reflex, pure tone audiometry, transient evoked (TEOAEs) and distortion product (DPOAEs) otoacoustic emissions on day 1 and on day 22 of treatment with vincristine. TEOAEs were analyzed in terms of emission level and reproducibility as a function of frequency. DPOAEs were obtained as DP-grams and were analyzed in terms of amplitude. RESULTS: The analyzed parameters of TEOAEs and DPOAEs revealed a declining tendency, although changes did not reach statistical significance. Pure tone audiometry and stapedial reflex thresholds were not altered. CONCLUSION: For the population of this study, vincristine did not seem to cause significant alterations of otoacoustic emissions' recordings and consequently significant outer hair cell damage. PMID- 16359738 TI - Sequence polymorphisms in the receptor-binding domain of Plasmodium falciparum EBA-175: implications for malaria vaccine development. PMID- 16359739 TI - Safety and low molecular weight heparin in older people in a hospital with ambulatory care. AB - To examine major bleeding and mortality rates of low molecular weight heparin (LMWH) and unfractionated heparin (UFH) for patients with pulmonary embolism (PE) and/or deep vein thrombosis (DVT), a retrospective review of the medical records for 286 patients who presented at a local hospital with PE and/or DVT during the period November 2002-August 2003 was performed. DATA COLLECTED: presence of co morbidities, concurrent medications, presence, site and severity of bleeding, outcome. Of all the patients, 50.7% received LMWH plus warfarin, 21.0% received UFH plus LMWH plus warfarin, 14.0% received UFH and warfarin, and 9.8% received LMWH only. There were nine minor bleeds and six major bleeds, which resulted in four deaths. Being a hospitalized patient and being age > or =70 years were associated with a major bleed (p<0.05). For hospital inpatients age > or =70 years on UFH and LMWH the number of major bleeds/1000 patient days was 18.9 and 9.2, respectively. The major bleeding rate is comparable if not better than that reported in the literature in our hospital setting where nearly half of the anticoagulation services were provided as ambulatory care. The increased rate of bleeding in the elderly we found is consistent with the findings of previous studies. PMID- 16359740 TI - Functional outcome of elderly hip fracture patients: does diabetes matter? AB - The purpose of this study was to investigate whether diabetes mellitus may affect the functional outcome of hip fractured patients. We studied 759 consecutive patients admitted for hip fracture rehabilitation, out of whom 18.2% were diabetics. The functional outcomes of diabetics and nondiabetics were assessed by the functional independence measurement scale (FIM) at admission and discharge. Data were analyzed by t-tests, Pearson correlation, and chi-square test as well as by multiple logistic regression analysis. Compared with nondiabetics, diabetic patients were slightly younger (p=0.003) and more hyperlipidemic (p=0.01), had a higher prevalence of previous stroke (p=0.03) and lower cognitive Mini-Mental State Examination (MMSE) scores (p=0.007). Absolute and relative FIM parameters, at admission and discharge, were similar in both groups. A multiple logistic regression analysis showed that diabetes was independently, and inversely, associated with male gender [odds ratio (OR), 2.11 (95% CI, 1.41-3.18)] and higher admission motor-FIM [OR, 1.05 (95% CI, 1.03-1.07)], whereas higher cognitive scores upon admission emerged as being "protective" for being in the motor-FIM gain <20 [OR, 0.94 (95% CI, 0.91-0.97)]. However, diabetes was not associated with any of the parameters indicating unsuccessful rehabilitation. The findings suggest that there is no difference in the functional outcome of diabetic and nondiabetic patients, presenting for rehabilitation after surgery of hip fractures. Diabetes should not be considered as adversely affecting rehabilitation of such patients. PMID- 16359742 TI - Admission blood glucose and adverse outcomes in non-diabetic patients with myocardial infarction in the reperfusion era. AB - AIMS: To investigate the association between admission blood glucose levels and adverse outcomes after an incident acute myocardial infarction across a broad range of glucose levels in non-diabetic patients treated with modern therapy. METHODS: The relationship between admission blood glucose and 28-day case fatality was studied in 1348 consecutively hospitalized patients with a first ever myocardial infarction between January 1998 and December 2002 recruited from a population-based myocardial infarction registry. RESULTS: Patients were divided into quartiles on the basis of admission glucose level. Patients with elevated admission blood glucose had more adverse baseline characteristics than patients with lower glucose levels. After multivariable adjustment the odds ratios (95% confidence interval) for 28-day case fatality among those in the second, third and fourth quartile in comparison to the first quartile were 1.55 (0.49-4.87), 3.21 (1.06-9.74), and 3.73 (1.28-10.92), respectively (p for trend=0.0054). Admission hyperglycemia was also associated with complications during hospital stay among 28-day survivors. CONCLUSION: The risk for major complications after an incident myocardial infarction was closely related to admission blood glucose concentrations near to or within the normal range, and certainly below the diabetic threshold. Thus, admission hyperglycemia still provides an early marker of bad prognosis after an acute myocardial infarction in an era of modern therapy. PMID- 16359741 TI - Effects of folic acid supplementation on psychomotor performance and hemorheology in healthy elderly subjects. AB - Cognitive impairment is associated with increased blood concentrations of homocysteine and high blood viscosity. Previous studies have shown that vitamin B supplementation reduces homocysteine and enhances cognitive function in patients with mild dementia and low serum folic acid. However, whether folic acid enhances cognitive function in elderly subjects without dementia and normal serum folic acid is unknown. Twenty-four healthy elderly subjects (age 73.0+/-5.6 years, mean+/-S.D.) with normal serum folic acid (6.3+/-2.4 microg/l) and Mini Mental State Examination (MMSE) >27/30 were randomized to 4-week treatment with folic acid 5mg/day or placebo in a randomized, placebo-controlled, parallel-group study. Continuous Attention Test (CAT), Four-Choice Reaction Time (FCRT), Digit Symbol Substitution (DSS), Scanning Memory Sets (SMS), and blood viscosity for different shear rates were measured before and after treatment. Folic acid supplementation induced a significant increase in serum folic acid levels (+13.8 versus +1.6 microg/l, p<0.001) and fall in homocysteine levels (-1.91 versus 0.41 micromol/l, p=0.05) compared to placebo. However, there was no significant change in CAT, FCRT, DSS, SMS, and blood viscosity between the two groups. Short term folic acid supplementation does not enhance psychomotor performance or reduce blood viscosity in healthy elderly subjects with normal serum folic acid levels and preserved cognitive function. PMID- 16359743 TI - Real-time in vivo dosimetry using micro-MOSFET detectors during intraoperative electron beam radiation therapy in early-stage breast cancer. AB - PURPOSE: In a previous paper we reported the results of off-line in vivo measurements using radiochromic films in IOERT. In the present study, a further step was made, aiming at the improvement of the effectiveness of in vivo dosimetry, based on a real-time check of the dose. MATERIALS AND METHODS: Entrance dose was determined using micro-MOSFET detectors placed inside a thin, sterile, transparent catheter. The epoxy side of the detector was faced towards the beam to minimize the anisotropy. Each detector was plugged into a bias supply (standard sensitivity) and calibrated at 5 Gy using 6 MeV electrons produced by a conventional linac. Detectors were characterized in terms of linearity, precision and dose per pulse dependence. No energy and temperature dependence was found. The sensitivity change of detectors was about 1% per 20 Gy accumulated dose. Correction factors to convert surface to entrance dose were determined for each combination of energy and applicator. From November 2004 to May 2005, in vivo dosimetry was performed on 45 patients affected by early-stage breast cancer, who underwent IOERT to the tumour bed. IOERT was delivered using electrons (4-10 MeV) at high dose per pulse, produced by either a Novac7 or a Liac mobile linac. RESULTS: The mean ratio between measured and expected dose was 1.006+/-0.035 (1 SD), in the range 0.92-1.1. The procedure uncertainty was 3.6%. Micro-MOSFETs appeared suitable for in vivo dosimetry in IOERT, although some unfavourable aspects, like the limited lifetime and the anisotropy with no build-up, were found. Prospectively, a real-time action level (+/-6%) on dose discrepancy was defined. CONCLUSIONS: Excellent agreement between measured and expected doses was found. Real-time in vivo dosimetry appeared feasible, reliable and more effective than the method previously published. PMID- 16359744 TI - Ocular adnexal mucosa-associated lymphoid tissue lymphoma treated with radiotherapy. AB - Forty-two patients with stage IE ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma were retrospectively analyzed. Five-year local control and progression-free survival rates were 100 and 77%, respectively. The most common relapsed site was the contralateral orbit. Thirty Gy of local irradiation seemed to be quite effective and safe. PMID- 16359745 TI - Attenuated cortisol response to alcohol in heavy social drinkers. AB - Individual differences in response to stress may play a role in the development and maintenance of addictive behaviors. While there is evidence that people with a biological family history for alcoholism have a blunted cortisol response to alcohol, data are lacking in other at-risk subgroups, such as heavy social drinkers. The present study examined salivary cortisol response to administration of 0.0, 0.4 (2 drink equivalent), and 0.8 g/kg (4 drink equivalent) alcohol in two groups of social drinkers: heavy drinkers (n=32) and light social drinkers (n=23). The study was conducted double-blind and drink-order was counterbalanced between groups. Salivary cortisol and subjective measures were obtained at predrink baseline, and 15, 45, 105, and 165 min after beverage consumption. Results showed a significant groupxdosextime interaction (p<0.005), with alcohol (0.8 g/kg) producing an attenuated cortisol response in heavy drinkers compared to the light drinkers during the declining phase of the BAC. This outcome remained even after controlling for the effects of smoking status, family history of alcoholism, sex, and negative affect ratings during the session. Neither placebo nor the lower dose of alcohol significantly increased cortisol levels. In sum, a relatively high dose of alcohol produced a smaller increase in cortisol in heavy drinkers compared to light drinkers. The reduced cortisol reactivity in the heavier drinkers is consistent with reports that individuals at risk for alcoholism are hyporesponsive to physical and psychological stress. Further research may help determine whether alteration in cortisol response to alcohol is a biological marker of the propensity to abuse alcohol. PMID- 16359746 TI - Secretion of functional human interleukin-3 from Bacillus subtilis. AB - The Gram-positive bacterium Bacillus subtilis is well-known for its huge capacity to produce secreted bacterial enzymes. Nevertheless, the secretion of pharmaceutically interesting recombinant proteins by this organism is frequently inefficient. This paper documents for the first time on the optimisation of B. subtilis for the production of human interleukin-3 (hIL-3), a four-helix bundle cytokine, which stimulates the proliferation and differentiation of a broad range of blood cells. By developing a host-vector system on the basis of the multiple protease-deficient B. subtilis strain WB700 and a multicopy plasmid containing two tandemly positioned strong promoters plus an efficient signal sequence, the hIL-3 protein was efficiently produced and secreted into the growth medium. As verified by SDS-PAGE, mass spectrometry and cross-linking experiments with a thiol-specific reagent, intact and properly folded hIL-3 was purified from the B. subtilis growth medium. Bioactivity tests showed that the isolated hIL-3 was able to specifically induce proliferation of the hIL-3-dependent leukaemia cell line MO7e. Using the eight-fold protease-deficient strain WB800 the hIL-3 accumulation in the growth medium was increased to levels up to 100 mg l(-1). PMID- 16359747 TI - Nitric oxide mediates inactivation of glutathione S-transferase in suspension culture of Taxus cuspidata during shear stress. AB - The importance of nitric oxide (NO) in regulating plant cell responses to environmental stresses is becoming evident. Here the possible role of NO in suspension cultures of Taxus cuspidata under shear stress was investigated in a Couette-type shear reactor. It was found that shear stress with 190 s(-1) caused NO generation in 8 h. NO formation can be inhibited by N-nitro-L-arginine, a nitric oxide synthase inhibitor. Moreover, the activity of glutathione S transferase (GST), a principal enzyme responsible for detoxification, decreased during shear stress. This inactivation partially recovered when NOS inhibitor or NO scavenger was added into cell cultures during shear stress. Treatment with reactive nitrogen species (RNS) also caused inactivation of GST in cells. The results indicate that NO plays a crucial role in GST inactivation in Taxus cuspidata cells under shear stress. PMID- 16359748 TI - Update on HAART in HIV. AB - Highly active antiretroviral combination therapy (HAART) has been responsible for a dramatic decrease in AIDS mortality since 1996, and has changed the clinical profile of HIV infection from a sub-acute lethal to a chronic ambulatory disease. HAART consists of a double nucleoside (NRTI) backbone plus either a non nucleoside reverse transcriptase inhibitor (NNRTI) or a ritonavir pharmacologically enhanced protease inhibitor (PI/r). Triple NRTI combinations are less potent than 2NRTIs/NNRTI or 2NRTIs/PI/r combinations. Antiretroviral first-line therapy is rapidly moving towards more convenient and less toxic regimens. Three double NRTI co-formulations are now available, and the risk of mitochondrial toxicity is low with drugs such as 3TC, FTC and tenofovir. Similarly, atazanavir, a recently available PI, can be given once daily and is less metabolically toxic than other PIs. Antiretroviral salvage therapy takes advantage of the development or availability of new drugs, either from existing (tipranavir, TMC 114 as new PIs) or new classes (T20 as a fusion inhibitor), that remain active on many triple-class drug resistant viruses. More progress is needed in the field of drug discovery, since a significant proportion of patients still die from AIDS with a multi-resistant virus, and since the incidence of primary HIV resistance is increasing in various parts of the world. PMID- 16359749 TI - A similarity measure for case based reasoning modeling with temporal abstraction based on cross-correlation. AB - Adverse drug events (ADEs) are a major limitation of drug safety. They are often caused by inappropriate selection of dose and the concurrent use of drugs modulating each other (drug interaction). Risk assessment and prevention strategies must therefore consider co-administered drugs, individual doses, and their timing. In a new approach we evaluated the performance of cross correlation, commonly used in signal processing, to determine similarities in patient treatments. To achieve this, patient treatments were modeled as groups of vectors representing discrete time intervals. These vectors were cross-correlated and the results evaluated to find clusters in time courses indicating similarity in treatment of different patients. To evaluate our algorithm, we then created a number of test cases. The focus of this article is on each treatment, and its pattern in time and dosage. The algorithm successfully produces a relatively low similarity score for cases that are completely different with respect to their pattern of time and dosage but high scores when they are equal (score of 0.699) or similar (score of 0.528) in their therapies, and thus succeeds in having a relatively high specificity (27/30). Such an approach might help to considerably reduce the problem of false alarms which hampers most existing alerting systems for medication errors or impending ADEs. PMID- 16359750 TI - Erlotinib in patient with non-small cell lung cancer: significant symptomatic improvement within a few days but without change in chest X-ray. PMID- 16359751 TI - Intratumoral administration of cisplatin through a bronchoscope followed by irradiation for treatment of inoperable non-small cell obstructive lung cancer. AB - PURPOSE: Patients presenting with inoperable non-small cell carcinoma of the lung associated with severe bronchial obstruction are at a high risk for developing post-obstructive pneumonia or respiratory failure. This often leads to death in weeks to months. Several studies suggest that initial use of debulking of obstructed airways by Nd-YAG laser photo resection or by cryotherapy lessens morbidity by reducing infections and respiratory insufficiencies. This can shorten hospitalization, improve the quality of life, and prolong survival. It has also been demonstrated that patients first treated for debulking and then by irradiation have better survival than similar patients treated with irradiation alone. Intratumoral (IT) injection of cytotoxic drugs (IT chemotherapy) has also been successfully used to debulk airways. The aim of the present work was to study the effectiveness, safety, and feasibility of initial debulking by IT chemotherapy with cisplatin combined with irradiation with a curative intent in the treatment of obstructive inoperable non-small cell lung cancer. PATIENTS AND METHODS: Twenty three patients were treated first by bronchoscopic IT injection of up to 40 mg cisplatin solution (4 mg/ml), administered weekly four times (on days: 1, 8, 15, 22) and then by irradiation. RESULTS: At the end of a 3-week period of IT treatment, 11 of 23 patients showed an increase in airway lumen diameter of more than 50% (good response), 8 patients showed an improvement of 25 50% (partial response) and 4 patients showed an increase of less than 25% (small response). Overall, debulking by IT chemotherapy was considered clinically effective in 19 of 23 patients and only marginally effective in 4 patients. Statistical analysis indicated a statistically significant improvement in lumen diameter (P < 0.001) for all patients. The combination of IT chemotherapy and irradiation resulted in prolonged survival in the patients with good response (median survival 636 days) compared to patients with partial response (median survival 342 days). The four patients with only a small response had a median survival of 202 days. CONCLUSIONS: Debulking by intratumoral injection of cisplatin was shown to be a safe, simple, and cost effective procedure. There were no severe side effects and complications. Initial debulking of obstructed airways by IT administration of cisplatin, followed by irradiation requires further studies in order to determine its effect on survival. PMID- 16359752 TI - Laboratory measurement of cumulative food intake in humans: results on reliability. AB - The universal eating monitor (UEM) of Kissileff for laboratory measurement of food intake was modified and used with a newly developed special software to compute cumulative intake data. To explore the measurement precision of the UEM an investigation of test-retest-reliability of food intake parameters was conducted. The intake characteristics of 61 normal weight males and females were measured repeatedly in the laboratory with a measurement interval of 1 week. Yogurt of preferred flavor served as test meal. Test-retest-reliability of intake characteristics ranged from .64 (number of bites) to .83 (bite-size) and was .73 for initial eating rate, but only .16 for change of eating rate. Sex, BMI and eating habits according to TFEQ-factors had no significant effects on reliability of intake characteristics. The retest-reliability for all but one of the laboratory intake measures is comparable to standards of personality questionnaires and seems not to be influenced by sex or trait characteristics of eating behavior. PMID- 16359753 TI - Virus diffusion in isolation rooms. AB - In hospitals, the ventilation of isolation rooms operating under closed-door conditions is vital if the spread of viruses and infection is to be contained. Engineering simulation, which employs computational fluid dynamics, provides a convenient means of investigating airflow behaviour in isolation rooms for various ventilation arrangements. A cough model was constructed to permit the numerical simulation of virus diffusion inside an isolation room for different ventilation system configurations. An analysis of the region of droplet fallout and the dilution time of virus diffusion of coughed gas in the isolation room was also performed for each ventilation arrangement. The numerical results presented in this paper indicate that the parallel-directional airflow pattern is the most effective means of controlling flows containing virus droplets. Additionally, staggering the positions of the supply vents at the door end of the room relative to the exhaust vents on the wall behind the bed head provides effective infection control and containment. These results suggest that this particular ventilation arrangement enhances the safety of staff when performing medical treatments within isolation rooms. PMID- 16359754 TI - Immunocytochemical localization of adrenomedullin 2/intermedin-like immunoreactivity in human hypothalamus, heart and kidney. AB - Adrenomedullin 2/intermedin (AM2/IMD) is a novel member of the calcitonin/calcitonin gene-related peptide (CGRP) peptide family. AM2/IMD has a vasodilator action, and antidiuretic and antinatriuretic effects in mice. The aim of the present study is to clarify immunolocalization of AM2/IMD in human hypothalamus, heart and kidney obtained at autopsy. Immunocytochemistry showed AM2/IMD-immunoreactive cell bodies in the paraventricular and supraoptic nuclei of human hypothalamus. Both parvocellular and magnocellular cells in the paravetricular nucleus are immunostained with AM2/IMD. Immunostaining of serial sections showed co-localization of AM2/IMD-like immunoreactivity and vasopressin in the paraventricular nucleus. Myocardial cells of the heart and renal tubular cells were positively immunostained with AM2/IMD, whereas neither renal glomeruli nor vasculature in the heart and kidney were immunostained. Reverse-transcriptase polymerase chain reaction confirmed expression of AM2/IMD mRNA in the brain, pituitary, heart and kidney. The present study has shown the wide expression of AM2/IMD in human hypothalamus, heart and kidney, raising the possibility that this novel peptide may be related to the central and peripheral regulation of the circulation and water-electrolyte metabolism. PMID- 16359755 TI - Human pancreatic polypeptide has a marked diurnal rhythm that is affected by ageing and is associated with the gastric TFF2 circadian rhythm. AB - Normal circadian variations in vasoactive intestinal polypeptide, somatostatin, cholecystokinin and pancreatic polypeptide were measured to determine if these alter with aging and to identify gastrointestinal regulatory hormones that might control the dramatic diurnal variation in the gastric cytoprotective trefoil protein TFF2. Plasma pancreatic polypeptide concentrations showed a marked diurnal rhythm (p < 0.0001). Basal and postprandial pancreatic polypeptide concentrations increased with age (p < 0.01). The timing of the diurnal rhythm was consistent with pancreatic polypeptide inhibiting TFF2 secretion and there was a negative association between pancreatic polypeptide and TFF2 concentrations (p < 0.002). The much higher pancreatic polypeptide concentrations in older people will induce increased satiety that may contribute to 'anorexia of ageing'. These results identify potential therapies for treatment of gastric mucosal morbidity and age-associated loss of appetite. PMID- 16359756 TI - Characterization of NPY receptor subtypes Y2 and Y7 in rainbow trout Oncorhynchus mykiss. AB - We report the cloning and pharmacological characterization of two neuropeptide Y (NPY) receptor subtypes, Y2 and Y7, in rainbow trout (Oncorhynchus mykiss). These subtypes are approximately 50% identical to each other and belong to the Y2 subfamily of NPY receptors. The binding properties of the receptors were investigated after expression in human HEK-293 EBNA cells. Both receptors bound the three zebrafish peptides NPY, PYYa, and PYYb, as well as porcine NPY and PYY, with affinities in the nanomolar range that are similar to mammalian Y2. The affinity of the truncated porcine NPY fragments, NPY 13-36 and NPY 18-36 was markedly lower compared to mammalian and chicken Y2. This suggests that mammalian and chicken Y2 are unique among NPY receptors in their ability to bind truncated peptide fragments. The antagonist BIIE0246, developed for mammalian Y2, did not bind either of the two rainbow trout receptors. Our results support the proposed expansion of this gene family by duplications before the gnathostome radiation. They also reveal appreciable differences in the repertoire and characteristics of NPY receptors between fish and tetrapods stressing the importance of lineage specific gene loss as well as sequence divergence after duplication. PMID- 16359757 TI - Ca2+ signaling, mitochondria and sensitivity to oxidative stress in aging astrocytes. AB - Age-related changes in astrocytes that could potentially affect neuroprotection have been largely unexplored. To test whether astrocyte function was diminished during the aging process, we examined cell growth, Ca2+ signaling, mitochondrial membrane potential (DeltaPsi) and neuroprotection of NGF-differentiated PC12 cells. We observed that cell growth was significantly slower for astrocytes cultured from old (26-29 months) mice as compared to young (4-6 months) mice. DeltaPsis in old astrocytes were also more depolarized (lower) than in young astrocytes and old astrocytes showed greater sensitivity to the oxidant tert butyl hydrogen peroxide (t-BuOOH). ATP-induced Ca2+ responses in old astrocytes were consistently larger in amplitude and more frequently oscillatory than in young astrocytes, which may be attributable to lower mitochondrial Ca2+ sequestration. Finally, NGF-differentiated PC12 cells that were co-cultured with old astrocytes were significantly more sensitive to t-BuOOH treatment than co cultures of NGF-differentiated PC12 cells with young astrocytes. Together, these data demonstrate that astrocyte physiology is significantly altered during the aging process and that the astrocyte's ability to protect neurons is compromised. PMID- 16359758 TI - [Medical comorbidity in Alzheimer's disease: baseline characteristics of the REAL.FR Cohort]. AB - PURPOSE: Alzheimer's disease (AD) evolves over about ten years with cognitive decline that can be considered as linear. Comorbidities are frequent in geriatric population. The major objective of this study is to determine whether comorbidity influences natural history of AD. MATERIALS AND METHODS: This is a prospective, multicentric French study (REAL.FR) of a cohort of ambulatory patients suffering from AD from a mild to a moderately severe stage, with a Mini-Mental State between 10 and 26, and followed with a caregiver. We evaluated the comorbidities and they were quantified using the Charlson index. RESULTS: We analysed 579 AD patients enrolled between April 2000 and June 2002. Majority of patients were women (72%). Average age and MMS average score were respectively 77.4 +/- 7.1 and 20.1 +/- 4.5. Cardiovascular diseases were the most frequent comorbid conditions (34%), before sensorial handicap (23%), and neurological diseases (18%) apart from dementia. Four AD patients groups differed according to the comorbidities figures, from none to more than three (maximum 8). Average Charlson index was 1.5 +/- 0.9. CONCLUSION: The follow-up of the four AD patients groups, differentiated by the comorbidities figures, should allow to precise the influence of comorbidities on the AD evolution. Charlson index could be used to quantify the comorbidities in the cohort's follow-up. However, this index, validated in a cohort of cancer patients, show limits for its use in geriatric population. PMID- 16359759 TI - Effects of mushroom, Pleurotus eryngii, extracts on bone metabolism. AB - We performed the present study to investigate whether Pleurotus eryngii extracts (PEX) play a role in bone metabolism. PEX treatment showed increase in the alkaline phosphatase activity of the osteoblasts and in the osteocalcin mRNA expression from primary osteoblasts. PEX also increased the expression of the Runx2 gene, and the secretion of osteoprotegerin from the osteoblasts showed marked increases after treatment with PEX. In addition, PEX treatment decreased the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells and resorption areas. In vivo studies, using rats with ovariectomy-induced osteoporosis revealed that PEX alleviated the decrease in the trabecular bond mineral density. PMID- 16359760 TI - A novel peptide isolated from phage library to substitute a complex system for a vaccine against staphylococci infection. AB - Staphylococcus aureus is a major human pathogen and many of the strains are resistant to conventional antibacterial treatment. The bacteria cause disease largely due to the production of multiple toxins, whose synthesis is controlled by an RNA molecule termed RNAIII. The production of RNAIII is induced by quorum sensing systems, one of them containing the protein RNAIII activating protein (RAP). Here we show that we partially purified supernatant of S. aureus, used this fraction to vaccinate mice, and selected antibody-binding peptides by phage display. Mice were vaccinated with the various peptides and challenged with S. aureus. One of the binding peptide termed R13 induced a protective immune response. Western blot analysis showed that the anti-R13 antibodies specifically bind to native or recombinant RAP. Mice vaccinated with R13 were protected and protection was sustained for the duration of the 6-month study period. Our results show that R13 could be used as a long-term effective protective-peptide vaccine to prevent S. aureus infections and once again show that targeting the RAP quorum sensing system is an effective approach to preventing staphylococcal infections. In addition our studies show that selection of specific protective peptides by phage display using sera induced by complex antigens is a rapid and effective way to identify the protective antigen and select for a peptide vaccine. PMID- 16359761 TI - An exploratory study of the conditions important in facilitating the empowerment of midwives. AB - OBJECTIVE: to explore the conditions important in facilitating the empowerment of midwives, as judged by practising midwives. DESIGN: a cross-sectional, descriptive, national survey of nurses and midwives practising in Ireland was undertaken in 2002. The questionnaire used in the survey included the 'Understanding of Empowerment Scale'. PARTICIPANTS: the data come from a larger national study of empowerment among nurses and midwives carried out by the authors in 2001/2 (Scott et al., 2003). A random sample of 4050 nurses and midwives in Ireland was obtained from the Live Register of An Bord Altranais (Irish Nursing Board). There was an overall response rate of 46% (n = 1781). Of these, 1340 people completed questionnaires, as 441 of those who replied were not in practice at the time. Of those who completed the questionnaire, 7% (n = 95) were practising midwives. Their findings are reported in this paper. MEASUREMENTS AND FINDINGS: factor analysis using the Principal Axis Factoring method of extraction and an oblique (Direct Oblimin) rotation was carried out on the Understanding of Empowerment Scale. This suggested four factors or conditions important for the empowerment of midwives: control, support, recognition and skills. KEY CONCLUSIONS: these findings relating to the conditions that facilitate empowerment in midwifery reflect the professional distinctiveness of midwifery and take into account the specific role and working environment of the midwife. IMPLICATIONS FOR PRACTICE: the identification of the important conditions to facilitate empowerment in midwifery provides a framework with which to explore ways of building on strengths and addressing weaknesses within the current situation for midwives in Ireland and elsewhere. These suggestive findings offer an opportunity to further develop a tool to measure levels of these conditions necessary to facilitate empowerment in midwifery. PMID- 16359762 TI - Hand replantations following complete amputations at the wrist joint: first experiences in Hanoi, Vietnam. AB - Five completely amputated hands at the level of the wrist joint were replanted. The five patients were all men, ranging from 19 to 31 years and the amputations were all the result of knife injuries. All the revascularizations were successful. Secondary tenolysis was required in one case. Postoperative functional results compared to the uninjured hand were evaluated according to five main criteria: (1) the patient's overall satisfaction with the hand; (2) recovery of flexor and extensor function of the thumb and fingers; (3) recovery of thumb opposition; (4) recovery of sensitivity in the median and ulnar nerve distributions; (5) ability of the surviving hand to perform daily tasks. The results showed that, although the replanted hands were never functionally as good as the contralateral hand, the patients were able to use them satisfactorily for activities of daily living. Our results demonstrate that replantation following complete amputation at the wrist joint level as a result of a sharp injury is extremely worthwhile and has excellent potential for functional recovery. PMID- 16359763 TI - The utilization of free tendon transplantation. 1912. PMID- 16359764 TI - Carpal canal pressures after volar plating of distal radius fractures. AB - Elevated pressures within the carpal canal are known to occur after distal radius fractures. Controversy exists regarding prophylactic carpal tunnel release after open reduction with internal fixation of distal radius fractures. The purpose of this study was to determine the tissue pressures within the carpal canal after volar plating of distal radius fractures. This study was a prospective, observational, IRB approved research study. Ten consecutive patients undergoing volar plating of distal radius fractures were enrolled. After the distal radius fractures were reduced and fixed with volar plates, slit catheters were inserted into the carpal canals for continuous postoperative pressure monitoring for 24 hours. The maximum recorded pressure was 65 mmHg, which occurred in the only patient with fracture blisters. Peak pressures remained at 40 mmHg or less (range 16-40, mean 29) in all patients without fracture blisters. At the conclusion of data collection, all pressures were at 31 mmHg or less. No patient complained of median nerve dysfunction during the study period. Routine prophylactic carpal tunnel release is not recommended after volar plating of distal radius fractures based on these pressure recordings. PMID- 16359765 TI - Gender differences in religious practices, spiritual experiences and health: results from the US General Social Survey. AB - Even though a majority of Americans report having spiritual/religious beliefs, the role of different dimensions of spirituality/religiosity in health is not well understood. Moreover, given that the experience of spirituality/religiosity differs for men and women, it is possible that the strength of the association between spirituality/religiosity and health may also differ by gender. The purpose of this study is to examine the relationship between spirituality/religiosity and three markers of health and well-being, and any gender differences in these relationships. First, we test the hypothesis that engagement with formal religious institutions (i.e. public religious activity) will be more beneficial for men than for women and we examine the role of denominational affiliation in any observed differences. Second, we directly compare effects of three different kinds of religious activities (public and private religious activity and spiritual experience) on health and well-being. Data are from the 1998 US General Social Survey, a nationally representative sample of non-institutionalized adults. Participants were asked about the frequency of engaging in public and private religious activities and having spiritual experiences. Psychological distress, happiness and self-rated health were used as indicators of health and well-being. Results suggest that weekly public religious activity was significantly associated with better health and well-being. Furthermore, this relationship was stronger for men than women and was influenced by denominational affiliation. When public religious activity, private religious activity and spiritual experiences were considered simultaneously, public religious activity emerged as the most consistent predictor of health and well-being among men. Among women, both public religious activity and spiritual experiences maintained an independent association with the health and well-being. These results suggest that it may not be appropriate to generalize findings about the relationship between spirituality/religiosity and health from one form of spirituality/religiosity to another, across denominations, or to assume effects are uniform for men and women. PMID- 16359766 TI - Metabolic profiling by 13C-NMR spectroscopy: [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells. AB - Metabolic profiling is defined as the simultaneous assessment of substrate fluxes within and among the different pathways of metabolite synthesis and energy production under various physiological conditions. The use of stable-isotope tracers and the analysis of the distribution of labeled carbons in various intermediates, by both mass spectrometry and NMR spectroscopy, allow the role of several metabolic processes in cell growth and death to be defined. In the present paper we describe the metabolic profiling of Jurkat cells by isotopomer analysis using (13)C-NMR spectroscopy and [1,2-(13)C(2)]glucose as the stable isotope tracer. The isotopomer analysis of the lactate, alanine, glutamate, proline, serine, glycine, malate and ribose-5-phosphate moiety of nucleotides has allowed original integrated information regarding the pentose phosphate pathway, TCA cycle, and amino acid metabolism in proliferating human leukemia T cells to be obtained. In particular, the contribution of the glucose-6-phosphate dehydrogenase and transketolase activities to phosphoribosyl-pyrophosphate synthesis was evaluated directly by the determination of isotopomers of the [1' (13)C], [4',5'-(13)C(2)]ribosyl moiety of nucleotides. Furthermore, the relative contribution of the glycolysis and pentose cycle to lactate production was estimated via analysis of lactate isotopomers. Interestingly, pyruvate carboxylase and pyruvate dehydrogenase flux ratios measured by glutamate isotopomers and the production of isotopomers of several metabolites showed that the metabolic processes described could not take place simultaneously in the same macrocompartments (cells). Results revealed a heterogeneous metabolism in an asynchronous cell population that may be interpreted on the basis of different metabolic phenotypes of subpopulations in relation to different cell cycle phases. PMID- 16359768 TI - Depressed mood, index finger force and motor cortex stimulation: a transcranial magnetic stimulation (TMS) study. AB - The present study utilized transcranial magnetic stimulation (TMS) of the motor cortex to understand basic motor processes associated with depressive symptoms independent of cognitive requirements or diagnostic category. To assess the integrity of the basic cortical-spinal-motor circuit associated with depressed mood, TMS to the motor cortex was used to initiate motor evoked potentials (MEPs) in forearm EMG and force production measured in the right (dominant) index finger. While at rest, a group with more depressive symptoms showed less force response in the index finger following stimulations compared with a group endorsing less depressive symptoms. A negative correlation between force response in the index finger at baseline (rest) following stimulation and the Beck depression inventory indicated that depressive mood symptom elevations were associated with less response to stimulations. The results argue for a greater importance placed on the relationship between depressive mood symptoms and basic motor processes. PMID- 16359769 TI - Verbal-autonomic response dissociations as traits? AB - Dissociations between subjective and physiological responses to stress are of central interest in coping research. However, little is known about their stability across situations and time. Two experimental sessions - separated by 1 year - were conducted to examine cross-situational consistency and longterm stability of HR-derived and SCL-derived dissociation scores. In year 1, a speech stressor, the cold pressor and a video stressor (viewing of the speech video) were applied. In year 2, mental arithmetics, anagrams and a torture video were presented. Thirty-five students participated and HR, SCL and negative affect were recorded. For each stressor, standardized changes in negative affect were subtracted from changes in autonomic reactivity (HR and SCL, respectively). Dissociation scores were relatively consistent across the stressors with HR derived scores exceeding SCL-derived scores. Longterm-stability proved acceptable (r=.61, P<.001 for HR-derived and r=.40, P<.05 for SCL-derived scores). In sum, verbal-autonomic response dissociations show considerable cross-situational and temporal stability and thus might be considered as traits. PMID- 16359767 TI - Biochemical and immunochemical characterisation of human diadenosine triphosphatase provides evidence for its identification with the tumour suppressor Fhit protein. AB - We describe here the purification and characterisation of the human enzyme diadenosine triphosphatase isolated from human platelets and leukocytes, offering biochemical and immunochemical evidence to identify this enzyme with the novel tumour suppressor Fhit protein, a homodimer composed of approximately 17 kDa monomers. It catalyses the Mg(2+)-dependent hydrolysis of diadenosine triphosphate, Ap(3)A, to AMP+ADP. The fluorogenic substrate di-ethenoadenosine triphosphate, epsilon-(Ap(3)A), and Fhit antibodies were used for enzymatic and immunochemical characterisations, respectively. Human Ap(3)Aase presents a native molecular mass of approximately 32 kDa and no significant differences were found in K(m) values (2 microM), activating effects by Mg(2+), Ca(2+), and Mn(2+), optimum pH (7.0-7.2) or inhibition by Zn(2+) and diethyl pyrocarbonate between the human enzyme and the recombinant Fhit protein. Suramin is a very potent competitive inhibitor of both human Ap(3)Aase and Fhit protein with K(i) values in the range 20-30 nM. Both human and rat Ap(3)Aase activity co-purifies with Fhit immunoreactivity under gel filtration, ion-exchange and affinity chromatography. Homogeneous human Ap(3)Aase preparations analysed by SDS-PAGE and Western blot analysis with Fhit antibodies elicit immunochemical responses corresponding to a approximately 17 kDa polypeptide, indicating a dimeric structure for the enzyme Ap(3)Aase. The strong inhibition of Fhit enzyme by the drug suramin, supports the need to investigate the therapeutic potential of Fhit Ap(3)Aase mediated by its interaction with suramin or related drugs. PMID- 16359770 TI - Term-dependency of P2 receptor-mediated contractile responses of isolated human pregnant uterus. AB - OBJECTIVE: The aim of the study was to test the hypothesis that in the human uterus, the effectiveness of P2 receptor-mediated contractile responses is up regulated during pregnancy. STUDY DESIGN: Experiments were performed on myometrial samples obtained from women undergoing caesarean section at 28-30 weeks of pregnancy (3 women, Group 1), 32-34 weeks of pregnancy (6 women, Group 2) and 38-41 weeks of pregnancy (16 women, Group 3). Concentration-response relationships for a non-selective P2 receptor agonist, adenosine 5'-triphosphate (ATP), a selective P2X receptor agonist, alpha,beta-methylene-ATP (alpha,beta meATP), and a frequency-response relationship for non-adrenergic non-cholinergic (NANC) electrical field stimulation (EFS) were obtained using routine pharmacological organ bath technique. Effects of pyridoxalphosphate azophenyl 2',4'-disulphonic acid (PPADS, 10(-5) M), a P2 receptor antagonist, were also evaluated. Parametric Student's t-test, non-parametric Wilcoxon T-test, Mann Whitney U-test, two-way analysis of variance (ANOVA) and Krushkal-Wallis tests were used for statistical analysis. RESULTS: ATP (10(-6) to 3 x 10(-4) M), alpha,beta-meATP (10(-7) to 3 x 10(-5) M) and EFS (2-32 Hz) evoked contractions of isolated pregnant uterus in all three groups. Uterus responses to ATP were not correlated with the term of pregnancy while the amplitude of uterine contractions to alpha,beta-meATP and EFS was higher in full term pregnancy than in earlier pregnancy. PPADS antagonized uterus responses to alpha,beta-meATP and EFS, but not to ATP, in all three groups. CONCLUSION: P2X receptor-mediated contractions of human pregnant uterus to alpha,beta-meATP and EFS, but not to ATP, are increased with the progression of pregnancy. PMID- 16359771 TI - Pelvic lymphadenectomy for cervical cancer: extraperitoneal versus laparoscopic approach. AB - OBJECTIVE: To compare the extraperitoneal versus the laparoscopic technique in performing pelvic lymphadenectomy in a series of patients undergoing a radical vaginal hysterectomy for locally advanced cervical cancer. STUDY DESIGN: Retrospective study with 42 patients undergoing a radical vaginal hysterectomy for cervical cancer. Patients from group A (20 patients) had a laparoscopic lymph node dissection and patients belonging to group B (22 patients) had an extraperitoneal lymphadenectomy. Historical data, clinical and surgical characteristics, perioperative and post-operative complications were analyzed. Follow-up was conducted according to the oncologic requirements. RESULTS: No significant difference was observed between the two groups in terms of blood loss, post-operative pain, transfusions, hospital stay and post-operative hematomas. The extraperitoneal group (group B) significantly showed a reduced operating time, a greater number of nodes removed (p<0.05). The only lymphocyst occurred in group B. CONCLUSIONS: Extraperitoneal pelvic lymphadenectomy can be considered an adequate technique to complement radical vaginal operations for cervical cancer. PMID- 16359772 TI - Pituitary desensitization for eight weeks after the administration of two distinct gonadotrophin-releasing hormone agonists. AB - OBJECTIVE(S): The objective was to evaluate the duration of pituitary desensitization after the administration of 3.5 mg of triptorelin (T) and leuprolin (L) depot preparations in patients with endometriosis. STUDY DESIGN: Two groups of 30 patients received, on 21st day of the cycle, 3.75 mg i.m. of triptorelin (T group), and of leuprolin acetate (L group). From the first to the eighth week following gonadotrophin-releasing hormone agonists (GnRH-a) administration both groups underwent pelvic ultrasound and serum follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) evaluation. Statistical analysis was performed using the ANOVA test and the median test. A p-value < 0.05 was considered significant. RESULTS: Pituitary suppression was achieved from two to six and from two to seven weeks after the administration of 3.75 mg of leuprolin and triptorelin, respectively. FSH and LH serum levels were significantly higher in the L group than in the T group after the fourth week. CONCLUSIONS: Leuprolin and triptorelin depots (3.75 mg) promote satisfactory ovarian suppression lasting for six and seven weeks, respectively, after administration, with significantly different ambient levels of endogenous LH. PMID- 16359773 TI - Preoperative serum thrombopoietin levels are higher in patients with ovarian cancer than with benign cysts. AB - OBJECTIVE: To assess the diagnostic relevance of serum thrombopoietin (TPO) levels, we compared serum TPO levels between the patients with ovarian cancer and benign ovarian cysts. We also correlated serum TPO concentrations in ovarian cancer patients to know prognostic factors such as disease stage, tumor grade, histological subtype, and residual tumor mass. STUDY DESIGN: Preoperative serum TPO levels were measured in women with epithelial ovarian cancer (n=51) and women with benign ovarian cysts (n=25) using an enzyme-linked immunosorbent assay. RESULTS: The serum TPO concentration was significantly elevated in the ovarian cancer patients as compared to the benign patient controls. Platelet counts and TPO in the serum did not correlate in the ovarian cancer patients. However, using a TPO cutoff of 90 pg/ml and a CA125 cutoff of 30 units/ml, when both markers were elevated, the specificity was as high as 92%. CONCLUSION: Serum TPO concentrations may have a role in the diagnostic clinical setting for discerning benign from malignant ovarian tumors. PMID- 16359774 TI - Maternal and perinatal outcome of preeclampsia with an onset before 24 weeks' gestation. Audit in a tertiary referral center. AB - OBJECTIVE: Preeclampsia, with an onset before 24 weeks' gestation is a rare but severe condition in pregnancy with little data of maternal and perinatal outcome, particularly after expectant management. We therefore, evaluated pregnancy outcome in these women at our department where temporising management was introduced as the standard policy in early onset preeclampsia. STUDY DESIGN: We analysed retrospectively all consecutive women with preeclampsia, with an onset before 24 weeks' gestation, between 1 January 1993 and 31 December 2002 at a tertiary university referral center. RESULTS: Twenty-six pregnancies, of which two were twin pregnancies, resulted in 65% of the women in at least one major maternal complication: maternal death (n=1), HELLP syndrome (n=16), eclampsia (n=5) and pulmonary edema (n=4). Thirty percent of these women presented already with serious morbidity at admission. The median prolongation of the pregnancy was 24 days (range 3-46 days). The overall perinatal mortality was 82%: 19 fetal deaths and 4 neonatal deaths. CONCLUSION: Early onset preeclampsia, with an onset before 24 weeks' gestation, results in considerable maternal and perinatal morbidity and mortality. Therefore, expectant management should not be considered as a routine treatment option in these patients. PMID- 16359775 TI - Factors influencing farmers' concerns regarding bovine tuberculosis in wildlife and livestock around Riding Mountain National Park. AB - Despite intensive efforts over the last century to eradicate bovine tuberculosis (TB) in North America, several hotspots of infected wildlife and livestock remain, raising concerns that the disease will never be eradicated. The stress and frustration for a farmer caused by having a herd test positive for TB or living in an infected region can be substantial. The goal of this study was to investigate the concerns of farmers around Riding Mountain National Park (RMNP) regarding the presence of TB in wildlife and livestock and conduct an exploratory analysis of causal factors. Data were collected from 786 farmers within 50 km of RMNP using a mail-back questionnaire. Overall, farmers indicated a high level of concern toward diseases in both wildlife and cattle relative to other concerns. The spatial variables that had the greatest influence on TB concern were both the distance of farms to the RMNP boundary and distance of farms to previous cases of TB. The most important aspatial factor associated with high TB concern was the frequency with which farmers observed elk on their land. These results underscore the important differences between 'objective' measures of risk, such as epidemiological estimates of disease prevalence, and subjective measures of disease concern, such as risk perception and acceptability of management actions. Written responses suggest that concerns regarding disease may affect how farmers view wildlife on their land and their relationship with neighbouring protected areas. Management activities that reduce the frequency of elk interactions with farms, but also recognize the complex relationship that farmers have with wildlife and protected areas, will be most effective in mitigating farmer concern regarding this important problem. PMID- 16359776 TI - Heterogeneous photocatalysed degradation of two selected pesticide derivatives, triclopyr and daminozid in aqueous suspensions of titanium dioxide. AB - Heterogeneous photocatalysed degradation of two selected pesticide derivatives, triclopyr (1) and daminozid (2), has been investigated in aqueous suspensions of titanium dioxide by monitoring the change in substrate concentration employing the UV Spectroscopic analysis technique and depletion of Total Organic Carbon (TOC) content as a function of irradiation time. The degradation kinetics were studied under different conditions such as reaction pH, substrate and catalyst concentration, different types of TiO2 and in the presence of electron acceptors such as hydrogen peroxide (H2O2), potassium bromate (KBrO3) and ammonium persulphate (NH4)2S2O8 in addition to molecular oxygen. The degradation rates were found to be strongly influenced by all the above parameters. The photocatalyst Degussa P25 was found to be more efficient as compared with other photocatalysts. The pesticide derivative triclopyr (1) was found to degrade faster as compared to daminozid (2). An attempt was also made to identify the intermediate products formed during the photooxidation process using GC/MS analysis. Probable pathways for the formation of products have been proposed. PMID- 16359777 TI - Underwater video as a monitoring tool to detect change in seagrass cover. AB - To date seagrass monitoring has involved the removal of seagrass from its environment. In fragile or highly disturbed systems, monitoring using destructive techniques may interfere with the environment or add to the burden of disturbance. Video photography is a form of non-destructive monitoring that does not require the removal of seagrass or interference with the environment and has the potential to be a valuable tool in monitoring seagrass systems. This study investigated the efficacy of video photography as a tool for detecting change in seagrass cover, using the temperate Australian species Amphibolis antarctica (Labill.) Sonder ex Aschers. Using visual and random point estimates of seagrass cover from video footage, it was possible to determine the minimum sample size (number of random video frames) needed to detect change in seagrass cover, the minimum detectable change in cover and the probability of the monitoring design committing a Type II error. Video footage was examined at three scales: transects (m apart), sites (km apart) and regions (tens of km apart). Using visual and random point estimation techniques, a minimum sample size of ten quadrats per transect was required to detect change in uniform and variable seagrass cover. With ten quadrats it was possible to identify a minimum detectable change in cover of 15% for uniform and 30% for variable seagrass cover. Power analysis was used to determine the probability of committing a Type II error from the data. Region level data had low power, corresponding to a high risk of committing a Type II error. Site and transect level data had high power corresponding to a low risk of committing a Type II error. Based on this study's data, managers using video to monitor for change in seagrass cover are advised to use data from the smaller scale, for example, site and transect level data. By using data from the smaller scale, managers will have a low risk of incorrectly concluding there has not been a disturbance when one has actually occurred. PMID- 16359778 TI - A formal model for consensus and negotiation in environmental management. AB - Environmental management decisions typically lie at the interface of science and public policy. Consequently, these decisions involve a number of stakeholders with competing agendas and vested interests in the ultimate decision. In such cases, it is appropriate to adopt formal methods for consensus building to ensure transparent and repeatable decisions. In this paper, we use an environmental management case study to demonstrate the utility of a mathematical consensus convergence model in aggregating values (or weights) across groups. Consensus models are applicable when all parties agree to negotiate in order to resolve conflict. The advantage of this method is that it does not require that all members of the group reach agreement, often an impossible task in group decision making. Instead, it uses philosophical foundations in consensus building to aggregate group members' values in a way that guarantees convergence towards a single consensual value that summarizes the group position. We highlight current problems with ad hoc consensus and negotiation methods, provide justification for the adoption of formal consensus convergence models and compare the consensus convergence model with currently used methods for aggregating values across a group in a decision making context. The model provides a simple and transparent decision support tool for group decision making that is straightforward to implement. PMID- 16359779 TI - Elective nephron sparing surgery should become standard treatment for small unilateral renal cell carcinoma: Long-term survival data of 216 patients. AB - OBJECTIVES: Our experiences with elective nephron-sparing surgery (NSS) for renal cell carcinoma (RCC) in a consecutive series of 216 patients are presented. Clinicopathological features and long-term oncological outcome is compared to patients treated with radical nephrectomy (RN). METHODS: Between 1975 and 2002, NSS was performed in 488 patients; 311 of these patients had elective indications. Renal cell carcinoma was found in 241/311 patients (77.5%). Long term follow up data could be obtained in 216/311 patients. Cancer-specific survival was estimated using the Kaplan-Meier method. Cox's regression analysis and log-rank tests were used to evaluate independent predictive values of different clinicopathological features. Survival data of the 216 patients after NSS surgery were compared to 369 patients with small RCC treated with RN. RESULTS: After a mean follow up of 66 months (median 64 months) 29 (13.4%) of 216 patients treated with NSS had died, 4 of them (1.8%) tumour-related. Tumour recurrence was detected in 12 patients (5.6%). 204 patients (94.4%) were free of tumour at last follow-up. Cancer specific survival rates at 5 and 10 years for patients treated with NSS (RN) were 97.8% (95.5%) and 95.8% (84.4%). CONCLUSIONS: Elective NSS surgery provides optimal long-term outcome in patients with small localized RCC. Compared to RN, renal parenchyma is preserved without any disadvantage in survival rates. Consequently elective NSS should be accepted as gold standard for small renal tumours. PMID- 16359780 TI - Modular training for residents with no prior experience with open pelvic surgery in endoscopic extraperitoneal radical prostatectomy. AB - PURPOSE: To establish a teaching program for the performance of endoscopic extraperitoneal radical prostatectomy (EERPE) that would ascertain the safe and efficacious training of residents with no previous experience with open pelvic surgery. MATERIALS AND METHODS: The technique of EERPE was divided in 12 segments with 5 levels of difficulty. We thus designed a training program, where the resident learned the procedure in a mentor-defined schedule. During each educational EERPE, the trainee only performed the operative steps corresponding to his acquired skill level. The mentor performed the remaining parts of the EERPE, with the trainee assisting. The first 50 and consequent 100 cases performed by the residents were compared to the first 50 and last 100 cases (cases 521-621) performed by the mentor. RESULTS: Two residents with no prior experience with open pelvic surgery participated in the study, and required 43 and 38 procedures respectively, until they were considered to be competent. The initial 50 procedures performed completely independently by the residents had mean operative times of 176 and 173 minutes. There were 2 intraoperative rectal injuries (one patient developed recto-urethral fistula), and 1 hemorrhage, and 1 lymphocele, postoperatively. The positive margin rate for pT2 disease was 14.3 and 11.5%, and for pT3 tumors 38.8 and 29.1%, respectively. After an additional 100 procedures operated by the same residents, mean operative times were 142 and 146 minutes. There was one patient who needed a transfusion. Postoperative complications requiring re-intervention were 1 hemorrhage, 2 anastomotic leakages and 4 symptomatic lymphoceles. The positive margin rate for pT2 disease was 12.8% and 6.5%, and for pT3 tumors 33.3% and 26.3% respectively. No statistical significant differences were observed when comparing with the mentors cases. CONCLUSION: We have showed that residents with no prior experience in open surgery of the pelvis can adhere to the modular training scheme and successfully perform the EERPE procedure with similar risk of complications compared to the tutor. PMID- 16359781 TI - Prognostic factors for successful percutaneous tibial nerve stimulation. AB - OBJECTIVE: In sacral as well as tibial nerve stimulation test stimulation is the main prognostic factor for success. In our study we tried to identify prognostic patient characteristics to improve patient selection for neuromodulation therapy. METHODS: PTNS was applied to 132 patients in 8 study centers (51 men, 81 women, mean age of 53 years (range: 21-82)). 83 patients were treated for overactive bladder, 16 for non-obstructive urinary retention and 33 for chronic pelvic pain. All patients had to fill out micturition or pain diaries, as well as quality of life questionnaires before and after treatment. Patient characteristics were evaluated for their prognostic value for successful outcome of neuromodulation therapy with use of logistic regression. RESULTS: Objective success was seen in 32.6% of patients, subjective success in 51.5%. Most evaluated clinical parameters proved not to be of prognostic value. A history of sexual and/or physical abuse was found in 12 of 103 interviewed patients, but did not alter PTNS treatment outcome. However, a low total score at baseline in the SF-36 questionnaire proved to be predictive for not obtaining objective (OR 0.444 [95% CI: 0.198-0.996], p = 0.04) or subjective success (OR 0.424 [CI: 0.203-0.887], p = 0.02). Especially patients with a low SF-36 Mental Component Summary were prone to fail neuromodulation therapy: OR 0.123 (95% CI: 0.273-0.552), p = 0.006 for objective success. These patients also scored worse on disease-specific quality of life questionnaires, although they had no different disease severity compared to patients with good mental health. CONCLUSION: Bad mental health as measured with the SF-36 Mental Component Summary does not depend on symptom severity and is a negative predictive factor for success of percutaneous tibial nerve stimulation. It therefore might be used as a tool for better patient selection in neuromodulation therapy. PMID- 16359782 TI - 3Beta-alkyl-androsterones as inhibitors of type 3 17beta-hydroxysteroid dehydrogenase: inhibitory potency in intact cells, selectivity towards isoforms 1, 2, 5 and 7, binding affinity for steroid receptors, and proliferative/antiproliferative activities on AR+ and ER+ cell lines. AB - Type 3 17beta-hydroxysteroid dehydrogenase (17beta-HSD) is involved in the biosynthesis of the potent androgen testosterone (T), which plays an important role in androgen-sensitive diseases. In an attempt to design compounds to lower the level of T, we designed androsterone (ADT) derivatives substituted at the position 3beta as inhibitors of type 3 17beta-HSD, and then selected the eight most potent ones (compounds 1-8) for additional studies. In an intact cell assay, they inhibited efficiently the conversion of natural substrate 4-androstene-3,17 dione into T, although they were less active in intact cells (IC50 approximately 1 microM) than in homogenated cells (IC50=57-100 nM). A study of the inhibitory potency with four other 17beta-HSDs revealed they were selective, since they do not inhibit reductive types 1, 5 and 7, nor oxidative type 2. Interestingly, they did not show any binding affinity for steroid receptors (androgen, estrogen, glucocorticoid and progestin). Only two inhibitors, 3beta-phenyl-ADT (5) and 3beta-phenylmethyl-ADT (6) showed some proliferative activities on an AR+ cell line and on an ER+ cell line, but their effects were not mediated through the androgen or estrogen receptors. This study identified selective inhibitors of type 3 17beta-HSD acting through a mixed-type inhibition, and devoid of non suitable androgenic and estrogenic proliferative activities. The more potent inhibitors were 3beta-hexyl-ADT (2), 3beta-cyclohexylethyl-ADT (4) and 3beta phenylethyl-ADT (7). PMID- 16359783 TI - A study of the extraction behavior of thallium with tribenzylamine as the extractant. AB - A simple solvent extraction study for thallium has been conducted based on the extraction of thallium as a chlorocomplex (tetrachlorothallate (III) anion) with tribenzylamine. Radiolabelled Tl (III) was used to study the extraction behavior of thallium. The beta activity of the extracted ion-association complex has been monitored using liquid scintillation counting and Geiger Muller counter. The effect of various diluents such as toluene, chloroform and isobutylmethylketone (MIBK) were studied. The extraction efficiency was 95% when chloroform and MIBK were used as diluents for tribenzylamine at an overall concentration of 0.25 mol dm(-3) NaCl and 0.5 mol dm(-3) H(2)SO(4). The phenomenon of severe chemical quenching was observed with chloroform as the diluent. The recovery of thallium was found to be 95% at varying concentrations ranging from 5 to 20 microg ml(-1) Tl (III) with a relative standard deviation of 2.5%. PMID- 16359784 TI - Enhanced electrokinetic dissolution of naphthalene and 2,4-DNT from contaminated soils. AB - Electrokinetic soil remediation has been proven to remove heavy metals and polar organics from low hydraulic conductivity subsurface environment. In this study, carboxymethyl-beta-cyclodextrin (CMCD) was used as a carrier to assist electrokinetic treatment for removal of low polarity organic contaminants from soils (2.2% organic carbon content). Naphthalene and 2,4-dinitrotoluene (2,4-DNT) were selected as the test compounds. The results from columns experiments showed that 46 and 43% of naphthalene and 2,4-DNT, respectively, were removed using 0.01 N NaNO(3) flushing solution with 40 V electrical potential while 70 and 72% of naphthalene and 2,4-DNT were removed using 2 g/L CMCD solution. Naphthalene and 2,4-DNT removal was enhanced to 83 and 89%, respectively, by using 2 g/L CMCD with 40 V electrical potential. Findings from this study also demonstrated that cyclodextrin assisted electrokinetics can enhance the removal rate of naphthalene and 2,4-DNT. Electric potential applied has more influence on the contaminant removal than the amount of CMCD used. Higher voltage application caused increase in the removal rate of naphthalene and 2,4-DNT, and appeared to be one of the critical factors in obtaining higher contaminant removal while increasing CMCD solution concentration above 2 g/L appeared to have little effect on the contaminant removal. PMID- 16359785 TI - Removal of COD from landfill leachate by electro-Fenton method. AB - The treatment of landfill leachate by electro-Fenton (E-Fenton) method was carried out in a batch electrolytic reactor. The effect of operating conditions such as reaction time, the distance between the electrodes, electrical current, H(2)O(2) to Fe(II) molar ratio, Fenton's reagent dosage and H(2)O(2) feeding mode on the efficacy of E-Fenton process was investigated. It is demonstrated that E Fenton method can effectively degrade leachate organics. The process was very fast in the first 30 min and then slowed down till it was complete in 75 min. There exists an optimal distance range between the electrodes so that an over 7% higher chemical oxygen demand (COD) removal was achieved than the electrodes positioned beyond this range. COD removal efficiency increased with the increasing current, but further increase of current would reduce the removal efficiency. Organic removal increased as Fenton's reagent dosage increased at the fixed H(2)O(2) to Fe(II) molar ratio. COD removal was only 65% when hydrogen peroxide alone was applied to the electrolytic reactor, and the presence of ferrous ion greatly improved COD removal. COD removal efficiency increased with the increase of ferrous ion dosage at the fixed hydrogen peroxide dose and reached highest at the 0.038 mol/L of ferrous ion concentration. COD removal would decrease when ferrous ion concentration was higher than 0.038 mol/L. The stepwise or continuous addition of hydrogen peroxide was more effective than the addition of hydrogen peroxide in a single step. E-Fenton method showed the synergetic effect for COD removal as it achieved higher COD removal than the total COD removal by electrochemical method and Fenton's reagent. PMID- 16359786 TI - Decolorization kinetics of Procion H-exl dyes from textile dyeing using Fenton like reactions. AB - The decolorization kinetics of three commercially used Procion H-exl dyes was studied using a Fenton-like reagent. The effect of the major system parameters (pH, concentration of H(2)O(2) and Fe(3+) and initial dye concentration) on the kinetics was determined. For comparison, the effect of the use of UV irradiated Fenton-like reagent and of Fenton reagent on the kinetics was also examined. In addition, mineralization rates and the biodegradability improvement as well as the effect of the addition of Cl(-), CO(3)(2-) or HCO(3)(-) on the decolorization rates was studied. The reactions were carried out in a 300 ml stirred cylindrical reactor with the capability of UV irradiation. The dye half-life time goes through a minimum with respect to the solution pH between 3 and 4. It also exhibits a broad minimum with respect to Fe(3+) and H(2)O(2) at molar ratios of H(2)O(2)/Fe(3+) from about 100 to 10. The addition of CO(3)(2-) and HCO(3)(-) substantially reduces the decolorization rates, while this effect is significantly less pronounced with Cl(-). At an optimum range of parameters, the mineralization rate (TOC reduction) is very slow for the Fenton-like process (TOC decrease from an initial 49.5 to 41.1 mg/l after 30 min and to only 35.2 mg/l after 600 min), but it increases significantly for the photo-Fenton-like process (to TOC values of 39.7 and 11.4 mg/l, respectively). The biodegradability, as expressed by the BOD/COD ratio, increases significantly from an initial value of 0.11-0.55 for the Fenton-like and to 0.72 for the photo-Fenton-like processes. PMID- 16359787 TI - Phosphate removal from water by fly ash: factorial experimental design. AB - The influence of three variables (phophate concentration, initial pH of solution (pH(0)) and the fly ash dosage) on the removal efficiency of phosphate (% E) and equilibrium pH of solution (pH(eq)) by using fly ash was studied by means of 2(3) full factorial experimental designs. The parameters coded as x(1), x(2) and x(3), consecutively(,) were used. The parameters were investigated at two levels (-1 and 1). The effects of these factors on dependent variables, namely, % E and pH(eq) were investigated. To determine the significance of effects, the analysis of variance with 95% confidence limits was used. It was shown that % E and pH(eq) obtained in this study were found to be 99.6% and 11.16, corresponding to the operating condition of 25 mg l(-1), 2 g l(-1) and 5.5 for the phosphate concentration, fly ash dosage and pH(0), respectively. PMID- 16359788 TI - Removal of soluble organics from water by a hybrid process of clay adsorption and membrane filtration. AB - The removal of phenol and o-cresol from water by a hybrid process of clay adsorption and ultrafiltration (clay-UF) was studied. Batch adsorption equilibrium experiments showed that the amount of adsorption for phenol and o cresol decreased in the order kaolin>montmorillonite at an equilibrium pH (pH(e)) of 9.1. The clay-UF experiments were performed as a function of clay dose, solution pH, and transmembrane pressure. The role of pH in clay-UF process mainly depended on the acid-base nature of phenols and clays, and the charge of UF membrane. The rejection of phenol increased with increasing pH, and had a maximum at pH(e)=8.2 with kaolin but at pH(e)=9.1 with montmorillonite. The rejection of o-cresol also increased with increasing pH, and had a maximum at pH(e)=9.2 with kaolin but at pH(e)=10.2 with montmorillonite. Such differences between solute rejections depended on the pK(a) of the solutes, zeta potential of the clays, and surface charge of the membrane. The amount of soluble organics adsorbed onto the surface of membrane was negligible and the flux slightly decreased with increasing transmembrane pressure. PMID- 16359789 TI - Novel immobilization of titanium dioxide (TiO2) on the fluidizing carrier and its application to the degradation of azo-dye. AB - A photocatalyst was prepared by attaching TiO(2) powder (diameter, 50nm) in the sol state to fluidizing spherical ceramic carriers using a silicon binder. A high initial photocatalytic activity and strong attachment was obtained at a sintering temperature of 500 degrees C. An azo-dye (Orange-G) was used as the test contaminant to examine the photocatalytic effect of the new photocatalyst. The initial pseudo-first order degradation rate constant for Orange-G was 0.11min( 1). However, the photocatalytic activity doubled when boric acid was added to the silicon binder at a B to Si ratio of 106.5%. When sodium ethoxide was added to the silicon binder at a sodium ion to Si ratio of 15.0%, as much as 80% of the initial photocatalytic activity was maintained after the photocatalyst had been agitated at 180rpm for 300min. Adding both boric acid at a B/Si ratio of 106.5% and sodium ethoxide at a Na/Si ratio of 15% increased the photocatalytic activity and stability by three and four times, respectively. PMID- 16359790 TI - Increased sensory neuron apoptotic death 2 weeks after peripheral axotomy in C57BL/6J mice compared to A/J mice. AB - Peripheral nerve transection results in a disconnection of the neuron from its target. As a result, a series of metabolic changes occur in the cell body that may cause neuronal death, mainly by apoptotic mechanisms. Although neurons from neonatal animals are the most susceptible, peripheral, lesion-induced, neuronal loss also occurs in adults, and is particularly evident in mouse sensory neurons. However, differences in genetic background cause particular isogenic strains of mice to react unevenly to peripheral nerve lesion. In this work, we investigated the occurrence of apoptosis as well as the ultrastructural changes in the dorsal root ganglion sensory neurons and satellite cells of C57BL/6J and A/J mice 2 weeks after ipsilateral sciatic nerve transection at the mid-thigh level. C57BL/6J mice displayed a stronger sensory neuron chromatolytic reaction that resulted in an increased loss of neurons when compared with isogenic A/J mice (p<0.01). Additionally, most of the degenerating neurons displayed the classic features of apoptosis. These findings reinforced previous data obtained by the terminal-deoxynucleotidyl transferase nick-end labeling (TUNEL) technique. PMID- 16359791 TI - Increased survival and migration of engrafted mesenchymal bone marrow stem cells in 6-hydroxydopamine-lesioned rodents. AB - Parkinson's disease is characterized by the loss of dopaminergic neurons in the substantia nigra. Attempted replacement of these neurons by stem cells has proved inconclusive. Bone marrow mesenchymal stem cells (MSC) are multipotent, differentiating into a variety of cells, including neuron-like cells. We used the 6-hydroxydopamine (6-OHDA) animal model of Parkinson's disease to assess migration and differentiation of transplanted MSC. We found in rodents that transplanted MSC survive better in the 6-OHDA-induced damaged hemisphere compared to the unlesioned side. Moreover, contralaterally engrafted MSC migrated through the corpus callosum to populate the striatum, thalamic nuclei and substantia nigra of the 6-OHDA-lesioned hemisphere. In conclusion, we demonstrate that 6 OHDA-induced damage increases the viability of transplanted MSC and attracts these cells from the opposite hemisphere. PMID- 16359792 TI - Deep tissue inflammation upregulates neuropeptides and evokes nociceptive behaviors which are modulated by a neuropeptide antagonist. AB - Promising recent developments in the therapeutic value of neuropeptide antagonists have generated renewed importance in understanding the functional role of neuropeptides in nociception and inflammation. To explore this relationship we examined behavioral changes and primary afferent neuronal plasticity following deep tissue inflammation. One hour following craniofacial muscle inflammation ipsilateral as well as contralateral head withdrawal thresholds and ipsi- and contralateral hindpaw withdrawal thresholds were lowered and remained reduced for 28 days. Elevated levels of calcitonin gene-related peptide (CGRP) within the trigeminal ganglion temporally correlated with this mechanical allodynia. Inflammation also induced an increase in the number of CGRP and substance P (SP)-immunopositive trigeminal ganglion neurons innervating inflamed muscle but did not evoke a shift in the size distribution of peptidergic muscle afferent neurons. Trigeminal proprioceptive muscle afferent neurons situated within the brainstem in the mesencephalic trigeminal nucleus did not express CGRP or SP prior to or following inflammation. Intravenous administration of CGRP receptor antagonist (8-37) two minutes prior to adjuvant injection blocked plasma extravasation and abolished both head and hindlimb mechanical allodynia. Local injection of CGRP antagonist directly into the masseter muscle prior to CFA produced similar, but less pronounced, effects. These findings indicate that unilateral craniofacial muscle inflammation produces mechanical allodynia at distant sites and upregulates CGRP and SP in primary afferent neurons innervating deep tissues. These data further implicate CGRP and SP in deep tissue nociceptive mechanisms and suggest that peptide antagonists may have therapeutic potential for musculoskeletal pain. PMID- 16359793 TI - Chronobiological characteristics of painful diabetic neuropathy and postherpetic neuralgia: diurnal pain variation and effects of analgesic therapy. AB - Clinical impressions suggest that neuropathic pain is often worse at night and significantly impairs sleep. However, the temporal pattern of neuropathic pain during waking hours has not been clearly characterized. Using clinical trial data, we have evaluated the diurnal variation of pain intensity before and during analgesic treatment in patients with diabetic neuropathy (DN) and postherpetic neuralgia (PHN). Pain intensity (0-10) measures throughout the day from a placebo controlled trial of around-the-clock administration of gabapentin, morphine and a gabapentin-morphine combination in neuropathic pain patients were examined. Baseline data in untreated patients revealed no effect of day of week but a significant effect of time of day in both DN (P < 0.001) and PHN (P < 0.001) such that pain intensity progressively increases throughout the day. This temporal pattern is essentially preserved during treatment with gabapentin, morphine and their combination. Neuropathic pain intensity progressively increases throughout the day and this temporal profile appears to be unaffected by treatment with gabapentin and/or morphine. Advancing our understanding of the chronobiology of neuropathic pain may shed new light on various neurohormonal and neurophysiologic influences and lead to the identification of novel therapeutic targets. Furthermore, recognizing diurnal pain patterns may guide treatment strategies such as the targeted timing of analgesic therapies. PMID- 16359794 TI - Cold sensitivity in axotomized fibers of experimental neuromas in mice. AB - Cold allodynia is a common complaint in patients with peripheral neuropathies. However, cold sensitivity of the different types of sensory afferents present in injured nerves is poorly known. We recorded activity evoked by cold in intact sensory fibers of the skin-saphenous nerve preparation and in axotomized sensory fibers of approximately 21 days-old neuromas of the saphenous nerve of mice, in vitro. Sixteen percent of the axotomized units responded to cooling with an accelerating discharge, which stopped immediately during rewarming. This response was similar to that observed in the intact cold-sensitive fibers. Temperature threshold distribution was broad in intact and axotomized cold fibers (30.7-22 degrees C and 34.5-14.5 degrees C, respectively). One-third of the axotomized cold-sensitive fibers were mechanosensitive and none of them displayed spontaneous activity at baseline temperature. In contrast, 33% of intact cold sensitive fibers exhibited low rates of ongoing discharges. In 60% of the cold sensitive, axotomized units, cold threshold was shifted towards warmer values by the TRPM8 agonist L-menthol. Seventy percent of axotomized, cold-insensitive units developed sensitivity to cold when exposed to 4-aminopyridine and their mean cold threshold (approximately 28 degrees C) was unaffected by menthol. Their response properties differed greatly from those of cold-sensitive units. In conclusion, the transducing capacity to cold stimuli is substantially recovered in neuromas. Furthermore, axotomized fibers maintain the 4-AP-sensitive, voltage activated, transient potassium conductance that counteracts the depolarizing effects of cold in the majority of intact, cold-insensitive primary afferents. Our results indicate that injured nociceptors do not develop abnormal cold sensitivity, suggesting that other mechanisms underlie the cold-induced allodynia following peripheral nerve injury. PMID- 16359795 TI - The relationship of adult attachment to emotion, catastrophizing, control, threshold and tolerance, in experimentally-induced pain. AB - Although insecure attachment has been associated with a range of variables linked with problematic adjustment to chronic pain, the causal direction of these relationships remains unclear. Adult attachment style is, theoretically, developmentally antecedent to cognitions, emotions and behaviours (and might therefore be expected to contribute to maladjustment). It can also be argued, however, that the experience of chronic pain increases attachment insecurity. This project examined this issue by determining associations between adult attachment characteristics, collected prior to an acute (coldpressor) pain experience, and a range of emotional, cognitive, pain tolerance, intensity and threshold variables collected during and after the coldpressor task. A convenience sample of 58 participants with no history of chronic pain was recruited. Results demonstrated that attachment anxiety was associated with lower pain thresholds; more stress, depression, and catastrophizing; diminished perceptions of control over pain; and diminished ability to decrease pain. Conversely, secure attachment was linked with lower levels of depression and catastrophizing, and more control over pain. Of particular interest were findings that attachment style moderated the effects of pain intensity on the tendency to catastrophize, such that insecurely attached individuals were more likely to catastrophize when reporting high pain intensity. This is the first study to link attachment with perceptions of pain in a pain-free sample. These findings cast anxious attachment as a vulnerability factor for chronic pain following acute episodes of pain, while secure attachment may provide more resilience. PMID- 16359796 TI - Deep brain stimulation for the alleviation of post-stroke neuropathic pain. AB - Our aim was to asses the efficacy of deep brain stimulation in post-stroke neuropathic pain. Since 2000, 15 patients with post-stroke intractable neuropathic pain were treated with deep brain stimulation of the periventricular gray area (PVG), sensory thalamus (Ventroposterolateral nucleus-VPL) or both. Pain was assessed using both a visual analogue scale and the McGill's pain questionnaire. VAS scores show a mean improvement of 48.8% (SD 8.6%). However, there is a wide variation between patients. This study demonstrates that it is an effective treatment in 70% of such patients. PMID- 16359797 TI - Acute low back pain: pain-related fear and pain catastrophizing influence physical performance and perceived disability. AB - Pain-related fear and pain catastrophizing are associated with disability and actual performance in chronic pain patients. In acute low back pain (LBP), little is known about the prediction of actual performance or perceived disability by pain-related fear and pain catastrophizing. This experimental, cross-sectional study aimed at examining whether pain-related fear and pain catastrophizing were associated with actual performance and perceived disability. Ninety six individuals with an episode of acute LBP performed a dynamic lifting task to measure actual performance. Total lifting time was used as outcome measure. The results show that pain-related fear, as measured with the Tampa Scale for Kinesiophobia, was the strongest predictor of this physical task. Using the Roland Disability Questionnaire as a measure of perceived disability, both pain related fear and pain catastrophizing, as measured with the Pain Catastrophizing Scale, were significantly predictive of perceived disability and more strongly than pain intensity was. The results of the current study suggest that pain related fear is an important factor influencing daily activities in individuals suffering an episode of acute LBP. The study results have important clinical implications, especially in the development of preventive strategies for chronic LBP. PMID- 16359798 TI - Referred pain and hyperalgesia in human tendon and muscle belly tissue. AB - The sensitivity of tendon and tendon-bone junction is not fully described although these tissues have high clinical impacts. This study assessed (1) pain intensity and referred pain caused by hypertonic saline injection to the proximal tendon-bone junction (PTBJ), tendon and muscle belly sites of tibialis anterior muscle and (2) pressure pain sensitivity, pre, during and post hypertonic saline injections. Eighteen subjects (14 males and 4 females) participated. Subjects also had constant mechanical stimulation for 120s at 130% of baseline pressure pain threshold (PPT) during which VAS parameters were recorded. VAS parameters after hypertonic saline for PTBJ (VAS area, VAS peak), and tendon sites (VAS area, duration and time to maximum VAS) were significantly (P < 0.05) higher than muscle belly. During hypertonic saline pain all three sites displayed local and frequently enlarged and referred pain areas. Hypertonic saline pain at the PTBJ and tendon transiently increased pressure sensitivity at these sites (P < 0.05). When referred pain was caused by mechanical stimulation it occurred predominantly at the PTBJ and tendon sites (86% cases). Constant mechanical stimulation caused steadily increasing pain (summation of pain) at all sites. Hypertonic saline pain at the tendon and PTBJ caused significantly higher (P < 0.001) final VAS scores compared to the muscle belly site. The results indicate the PTBJ and tendon sites are more sensitive and susceptible to sensitisation by hypertonic saline than muscle belly. Furthermore, there may be site specific central changes reflected by the differences in the results regarding sensitivity and summation over time. PMID- 16359799 TI - Primum non nocere. PMID- 16359800 TI - The role of developmental factors in predicting young children's use of a self report scale for pain. AB - Accurate pain assessment is the foundation for effective pain management in children. At present, there is no clear consensus regarding the age at which young children are able to appropriately use self-report scales for pain. This study examined young children's ability to use the Faces Pain Scale-Revised; (FPS R; [Hicks CL, von Baeyer CL, Spafford PA, van Korlaar I, Goodenough B. The Faces Pain Scale-Revised: toward a common metric in pediatric pain measurement. Pain 2001; 93: 173-83]) for pain in response to vignettes and investigated the role of developmental factors in predicting their ability to use the scale. One hundred and twelve healthy children (3-6 years old) were assessed for their ability to accurately use a common faces scale to rate pain in hypothetical vignettes depicting pain scenarios common in childhood. Accuracy was determined by considering whether children's judgements of pain severity matched the pain severity depicted in the various vignettes. Children were also administered measures of numerical reasoning, language, and overall cognitive development. Results indicated that 5- and 6-year-old children were significantly more accurate in their use of the FPS-R in response to the vignettes than 4-year-old children, who in turn were significantly more accurate than 3-year-old children. However, over half of the 6-year-olds demonstrated difficulties using the FPS-R in response to the vignettes. Child age was the only significant predictor of children's ability to use the scale in response to the vignettes. Thus, a substantial number of young children experienced difficulties using the FPS-R when rating pain in hypothetical vignettes, although the ability to use the scale did improve with age. PMID- 16359801 TI - Prevalence of Toxoplasma gondii antibodies in red deer (Cervus elaphus) and other wild ruminants from Spain. AB - Serum samples from 441 red deer (Cervus elaphus) and 161 other wild ruminant species, collected between 1993 and 2005 from six regions of Spain were tested for antibodies against Toxoplasma gondii by the modified agglutination test (MAT). Antibodies to T. gondii (MAT 1:25 or higher) were detected in 15.6% of red deer. Statistically significant differences were observed among sampling sites with seroprevalence in red deer from Catalonia (42.2%) being significantly higher compared with other Spanish regions (8.7%) (P < 0.05). Statistically significant differences were not observed between T. gondii seroprevalence and sex, age or management of hunting estates (open versus fenced). Seroprevalence of T.gondii infection in other ruminants species was 24% of 79 fallow deer (Dama dama), 21.8% of 33 in roe deer (Capreolus capreolus), 33.3% of three Spanish ibex (Capra pyrenaica), 20% of 10 chamois (Rupicapra pyrenaica), 10% of 10 barbary sheep (Ammotragus lervia), and 14.8% of 27 mouflon (Ovis ammon) in areas not including Catalonia, where no samples from these species were available. Serological results indicated a widespread exposure to T. gondii among wildlife in Spain and suggest that consumption of raw or inadequately cooked meat, as well as handling carcasses of wild game, should be taken into account as a source of infection for humans. PMID- 16359802 TI - World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines for evaluating the efficacy of ectoparasiticides against myiasis causing parasites on ruminants. AB - These guidelines have been prepared to assist in the planning, conduct and interpretation of studies for the assessment of efficacy of ectoparasiticides against the myiasis causing parasites of ruminants. These guidelines specifically focus on larvicidal efficacy against myiasis causing flies. Information is provided on the selection of animals, dose determination and dose confirmation studies, field studies, record keeping and result interpretation. These guidelines advocate the use of pen facilities for dose determination and dose confirmation studies for defining therapeutic and persistent efficacy. These guidelines are also intended to assist investigators on how to conduct specific experiments, to provide specific information for registration authorities involved in the decision-making process, to assist in the approval and registration of new ectoparasiticides, and to facilitate the world-wide adoption of standard procedures. PMID- 16359803 TI - World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines for evaluating the efficacy of ectoparasiticides against biting and nuisance flies on ruminants. AB - These guidelines have been prepared to assist in the planning, conduct and interpretation of studies for the assessment of the efficacy of ectoparasiticides (excluding repellents) against the biting and nuisance dipteran flies of ruminants. Information is provided on the selection of animals, dose determination and dose confirmation studies, field studies, record keeping and result interpretation. These guidelines advocate the use of pen facilities for dose determination and dose confirmation studies. These guidelines also are intended to assist investigators on how to conduct specific studies, to provide specific information for registration authorities involved in the decision-making process, to assist in the approval and registration of new ectoparasiticides, and to facilitate the worldwide adoption of standard procedures. PMID- 16359804 TI - World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines for evaluating the efficacy of ectoparasiticides against biting lice, sucking lice and sheep keds on ruminants. AB - These guidelines have been prepared to assist in the design, implementation and interpretation of studies for the assessment of the efficacy of ectoparasiticides against biting and sucking lice and sheep keds on ruminants. Information is provided on the selection of animals, dose determination, dose confirmation and field studies, record keeping and result interpretation. These guidelines advocate the use of pen facilities for dose determination and dose confirmation studies for defining therapeutic and persistent efficacy. These guidelines are also intended to assist investigators on how to conduct specific experiments, to provide specific information for registration authorities involved in the decision making process, to assist in the approval and registration of new ectoparasiticides, and to facilitate the world-wide adoption of standard procedures. PMID- 16359805 TI - Sensitivity of different Trypanosoma vivax specific primers for the diagnosis of livestock trypanosomosis using different DNA extraction methods. AB - There are several T. vivax specific primers developed for PCR diagnosis. Most of these primers were validated under different DNA extraction methods and study designs leading to heterogeneity of results. The objective of the present study was to validate PCR as a diagnostic test for T. vivax trypanosomosis by means of determining the test sensitivity of different published specific primers with different sample preparations. Four different DNA extraction methods were used to test the sensitivity of PCR with four different primer sets. DNA was extracted directly from whole blood samples, blood dried on filter papers or blood dried on FTA cards. The results showed that the sensitivity of PCR with each primer set was highly dependant of the sample preparation and DNA extraction method. The highest sensitivities for all the primers tested were determined using DNA extracted from whole blood samples, while the lowest sensitivities were obtained when DNA was extracted from filter paper preparations. To conclude, the obtained results are discussed and a protocol for diagnosis and surveillance for T. vivax trypanosomosis is recommended. PMID- 16359806 TI - A method for sheep scab control by applying selective treatment based on flock serology. AB - Sheep scab, caused by Psoroptes ovis, is a severe and debilitating disease that can be treated and controlled by the use of acaricidal dips or the use of broad spectrum avermectins. In Switzerland, control measures are state regulated. In particular, sheep should be routinely treated with ectocide dips or avermectin injections before they are moved onto common alpine pasture in late spring. However, a substantial part of the sheep population remains untreated and represents a potential reservoir for the mite population. Untreated sheep that are not moved to alpine pasture may infest treated sheep when flocks are reassembled in autumn. In an attempt to identify infested sheep, all flocks in the Canton of Schwyz (Switzerland) were serologically tested in 2001 and in 2002 (587 and 565 flocks, respectively). In 2003, a representative number (182 of the 531 flocks) was again investigated. Seropositive flocks were treated with doramectin (0.3 mg kg(-1) body weight, intramuscularly) from 2001 to 2003. In spring 2002, no chemo-methaphylaxis was given to seronegative flocks before movement onto common alpine pastures. Of the 587 flocks surveyed in spring 2001, 34 were seropositive (5.8%). These consisted of 21 infested with P. ovis, 1 with P. cuniculi, 4 with Chorioptes spp. and 8 of seropositivity of unknown origin; there was a decrease of seropositive flocks in spring 2002 (4.4%) with 15, 0, 2 and 8, respectively. Of the 182 flocks surveyed in spring 2003, just 4 flocks (2.2%) were seropositive. All the seropositive reactions in these flocks were the result of Chorioptes spp. infestations. There was a corresponding decrease in the proportion of seropositive animals from 6.3% in spring 2001 to 2.1% in spring 2003. These results corroborate the concept that it may be possible to target chemo-metaphylaxis and hence decrease the use of endectocides as well as of ectocides to control sheep scab. This would be of great benefit in reducing the likelihood of development of anthelmintic resistance against avermectins, decreasing the extent of environmental and human contamination with potentially toxic products and diminishing potential drug residues in meat and milk. PMID- 16359807 TI - Rapid changes in production and behavioral action of estrogens. AB - It is well established that sex steroid hormones bind to nuclear receptors, which then act as transcription factors to control brain sexual differentiation and the activation of sexual behaviors. Estrogens locally produced in the brain exert their behavioral effects in this way but mounting evidence indicates that estrogens also can influence brain functioning more rapidly via non-genomic mechanisms. We recently reported that, in Japanese quail, the activity of preoptic estrogen synthase (aromatase) can be modulated quite rapidly (within minutes) by non-genomic mechanisms, including calcium-dependent phosphorylations. Behavioral studies further demonstrated that rapid changes in estrogen bioavailability, resulting either from a single injection of a high dose of estradiol or from the acute inhibition of aromatase activity, significantly affect the expression of both appetitive and consummatory aspects of male sexual behavior with latencies ranging between 15 and 30 min. Together these data indicate that the bioavailability of estrogens in the brain can change on different time-scales (long- and short-term) that match well with the genomic and non-genomic actions of this steroid and underlie two complementary mechanisms through which estrogens modulate behavior. Estrogens produced locally in the brain should therefore be considered not only as neuroactive steroids but they also display many (if not all) functional characteristics of neuromodulators and perhaps neurotransmitters. PMID- 16359808 TI - Effects of environmental and pharmacological stressors on c-fos and corticotropin releasing factor mRNA in rat brain: Relationship to the reinstatement of alcohol seeking. AB - We have observed marked heterogeneity among different stressors in their ability to reinstate alcohol seeking in rats. Of the stressors we have tested, only the environmental stressor footshock and the pharmacological stressor yohimbine induce reinstatement. The reasons for such differences among stressors are not known. The purpose of the experiments presented here is to determine the neuroanatomical substrates that underlie these behavioral differences. To this end, we assessed whether stressors effective in inducing reinstatement of alcohol seeking activate a different set of neuronal pathways than do those that are ineffective, using the technique of in situ hybridization of the mRNAs for c-fos, a marker of neuronal activation, and corticotropin-releasing factor (CRF), a stress-related peptide we have shown to be critical to footshock-induced reinstatement of alcohol seeking. Exposure of rats to the environmental stressors footshock, restraint or social defeat, or the pharmacological stressors yohimbine or FG-7142 increased levels of the mRNAs for c-fos and CRF in the brain in a number of areas previously shown to be responsive to stressors. We found regionally specific effects of the stressors on c-fos and CRF mRNA in brain regions associated with the rewarding effects of alcohol and other abused drugs. The two stressors we have previously shown to be effective in inducing reinstatement of alcohol seeking, footshock and yohimbine, induced c-fos mRNA in the shell of the nucleus accumbens, and the basolateral and central amygdalar nuclei. These two stressors also induced CRF mRNA in the dorsal region of the bed nucleus of the stria terminalis. Taken together, these results provide evidence that activity in these regions may be involved in the reinstatement of alcohol seeking induced by these stressors. These results are also in keeping with the previously demonstrated role of CRF neurons in the dorsal bed nucleus of the stria terminalis in the reinstatement of alcohol seeking induced by stress. PMID- 16359809 TI - Activation of cerebral endothelium is required for mononuclear cell recruitment in a novel in vitro model of brain inflammation. AB - Brain inflammation is a common event in the pathogenesis of several neurological diseases. It is unknown whether leukocyte/endothelium interactions are sufficient to promote homing of blood-borne cells into the brain compartment. The role of mononuclear cells and endothelium was analyzed in a new experimental model, the isolated guinea-pig brain maintained in vitro by arterial perfusion. This preparation allows one to investigate early steps of brain inflammation that are impracticable in vivo. We demonstrate by confocal microscopy analysis that in vitro co-perfusion of pro-inflammatory agents and pre-activated fluorescent mononuclear cells induced endothelial expression of selectins and intracellular adhesion molecule-1 in correspondence of arrested mononuclear cells, and correlates with a moderate increase in blood-brain barrier permeability. Separate perfusion of pro-inflammatory agents and mononuclear cells induced neither mononuclear cell adhesion nor adhesion molecule expression. We demonstrate that co-activation of mononuclear cells and cerebral endothelium is an essential requirement for cell arrest and adhesion in the early stages of experimental cerebral inflammation. PMID- 16359810 TI - Working memory for order information: multiple cognitive and neural mechanisms. AB - Working memory for order information is mediated by different cognitive mechanisms that rely on different neural circuits. Here we discuss evidence that order memory involves mechanisms that range from general supervisory processes to process that maintenance fine-grained temporal position information. We suggest that neural regions-including the prefrontal cortex, motor cortex, parietal cortex and medial temporal structures-operate at different levels and processing stages to give rise to working memory for order information. PMID- 16359811 TI - Antisense-induced reduction in nucleus accumbens cyclic AMP response element binding protein attenuates cocaine reinforcement. AB - Repeated cocaine exposure up-regulates cyclic AMP signaling and increases the transcriptional activity of cyclic AMP response element binding protein (CREB) in the nucleus accumbens. To study the possibility that nucleus accumbens CREB activity regulates self-administration behavior, we tested the effects of a single, bilateral infusion of CREB antisense oligonucleotide into nucleus accumbens core and shell sub-regions on cocaine self-administration in rats. Nucleus accumbens core infusions of CREB antisense reduced CREB and the CREB regulated immediate early gene brain-derived neurotrophic factor by 31 and 27%, respectively, but failed to alter levels of the homologous CREB family proteins cyclic AMP response element modulator and activating transcription factor 1, and had no effect on CREB levels in adjacent nucleus accumbens shell tissue. Similar infusions of CREB antisense in either core or shell produced a transient downward shift in cocaine self-administration dose-response curves on a fixed ratio 5 (five responses/injection) reinforcement schedule, indicating a reduction in cocaine reinforcement that fully recovered 3 days after treatment. CREB antisense also increased the threshold dose of cocaine required for reinstating cocaine self-administration, indicating that nucleus accumbens CREB levels regulate the incentive properties of cocaine. When access to cocaine was less restricted on a fixed ratio 1 schedule, infusion of CREB antisense in the core, but not shell, caused a transient (1-2 days) reduction in stabilized cocaine self administration, but had no effect on responding maintained by sucrose pellets, indicating that basal CREB levels in the nucleus accumbens core regulate drug intake. None of these effects were produced by nucleus accumbens infusions of complementary sense oligonucleotide. These results suggest a necessary role for nucleus accumbens CREB activity in cocaine reinforcement, and, by converse analogy, up-regulation in CREB activity after chronic cocaine use could contribute to addiction-related increases in cocaine self-administration. PMID- 16359812 TI - Effects of a calcineurin inhibitor, tacrolimus, on glutamate-dependent potentiation in pelvic-urethral reflex in anesthetized rats. AB - Effects of tacrolimus, a protein phosphatase 2B inhibitor, on the reflex plasticity between the pelvic afferent nerve fibers and the urethra were examined in urethane-anesthetized rats. Repetitive stimulation (1 Hz) induced a potentiation (0.9+/-0.2 and 10.5+/-1.6 spikes in control and repetitive stimulation groups, respectively, P<0.01, N=10) in the activities of the pelvic urethral reflex. Intrathecal tacrolimus (0.1 mM, 10 microl, bolus) blocked repetitive stimulation-induced potentiation in pelvic-urethral reflex activities (3.2+/-0.9 spikes in tacrolimus group versus 10.5+/-1.6 spikes in repetitive stimulation group, P<0.01, N=10). Glutamate (intrathecal, 0.1 mM, 10 microl, bolus) and N-methyl-D-aspartic acid (intrathecal, 0.1 mM, 10 microl, bolus) both reversed the blocking effects exerted by tacrolimus on repetitive stimulation induced pelvic-urethral reflex potentiation (15.0+/-1.4 spikes in glutamate group and 11.4+/-1.4 spikes in N-methyl-D-aspartic acid group versus 3.2+/-0.9 spikes in tacrolimus-treated repetitive stimulation group, P<0.01, N=7). In addition, the reversal effect elicited by these two agonists of glutamate receptors showed no statistical difference (P=NS, N=7). All these results demonstrated that tacrolimus could block glutamatergic N-methyl-D-aspartic acid receptor-mediated potentiation in pelvic-urethral reflex activities. This finding may be pathologically relevant in patients who take tacrolimus as immunosuppressant therapy. Whether tacrolimus will induce urine incontinence in such patients or not needs further investigation. PMID- 16359813 TI - Human multidrug resistance protein 8 (MRP8/ABCC11), an apical efflux pump for steroid sulfates, is an axonal protein of the CNS and peripheral nervous system. AB - Dehydroepiandrosterone 3-sulfate and other neurosteroids are synthesized in the CNS and peripheral nervous system where they may modulate neuronal excitability by interacting with ligand-gated ion channels. For this modulatory activity, neurosteroids have to be locally released from either neurons or glial cells. We here identify the integral membrane protein ABCC11 (multidrug resistance protein 8) as an ATP-dependent efflux pump for steroid sulfates, including dehydroepiandrosterone 3-sulfate, and localize it to axons of the human CNS and peripheral nervous system. ABCC11 mRNA was detected in human brain by real-time polymerase chain reaction. Antibodies raised against ABCC11 served to detect the protein in brain by immunoblotting and immunofluorescence microscopy. ABCC11 was preferentially found in the white matter of the brain and co-localized with neurofilaments indicating that it is an axonal protein. Additionally, ABCC11 was localized to axons of the peripheral nervous system. For functional studies, ABCC11 was expressed in polarized Madin-Darby canine kidney cells where it was sorted to the apical membrane. This apical sorting is in accordance with the localization of ABCC11 to the axonal membrane of neurons. Inside-out plasma membrane vesicles containing recombinant ABCC11 mediated ATP-dependent transport of dehydroepiandrosterone 3-sulfate with a Km value of 21 microM. This transport function together with the localization of the ABCC11 protein in vicinity to GABAA receptors is consistent with a role of ABCC11 in dehydroepiandrosterone 3 sulfate release from neurons to sites of dehydroepiandrosterone 3-sulfate mediated receptor modulation. Our findings may provide a basis for the characterization of mutations in the human ABCC11 gene and their linkage with neurological disorders. PMID- 16359814 TI - Early establishment of adult-like nigrotectal architecture in the neonatal cat: a double-labeling study using carbocyanine dyes. AB - Virtually nothing is known about the ontogeny of substantia nigra, pars reticulata projections to the midbrain superior colliculus, even though this pathway is critical for the basal ganglia modulation of midbrain-mediated visuomotor behaviors. The present studies used the lipophilic carbocyanine dyes 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate and 1,1' dioctodecyl-3,3,3',3'-tetramethylindodi, 4-chlorobenzenesulfonate salt to examine the crossed and uncrossed nigrotectal projections in neonatal cats, from parturition to 14 days postnatal (the technical limits of the tracing technique). In retrograde experiments, paired placement of the dyes in each superior colliculus produced numerous retrogradely-labeled nigrotectal neurons, with the uncrossed neurons far out numbering their crossed counterparts. No double-labeled neurons were observed, indicating that crossed and uncrossed nigrotectal neurons are segregated at birth. In anterograde experiments, dye placements into each substantia nigra, pars reticulata resulted in an iterative series of labeled patches, aligned medial-to-lateral across the intermediate and deep superior colliculus, a pattern reminiscent of the adult. Uncrossed neonatal axons had simple linear morphologies with few branch points; by contrast, crossed axons displayed more extensive terminal arbors that were distributed diffusely throughout the rostrocaudal extent of the contralateral superior colliculus In the final series of experiments, one dye was placed unilaterally in the substantia nigra, pars reticulata, while the second dye was positioned in the predorsal bundle, in order to bilaterally label superior colliculus output neurons. Although both crossed and uncrossed axons appeared to have contacted superior colliculus output neurons, crossed axons preferentially targeted the soma and proximal dendrites, whereas uncrossed terminals were distributed more distally. Throughout this early postnatal period, no significant changes in cellular morphologies or gross modification of terminal projection patterns were observed; however, the presence of growth cones in even the oldest animals studied suggests that the refinement of the nigrotectal projections extends well into postnatal life. Nevertheless, the segregation of crossed and uncrossed nigrotectal neurons into a highly organized afferent mosaic that has established synaptic contacts with superior colliculus output neurons indicates that many of the salient features characterizing nigrotectal projections are established prior to the onset of visual experience. PMID- 16359815 TI - Spatial and temporal visual properties of single neurons in the suprageniculate nucleus of the thalamus. AB - The spatial and temporal visual sensitivity to drifting sinusoidal gratings was studied in 105 neurons of the suprageniculate nucleus of the feline thalamus. Extracellular single-unit recordings were performed in halothane-anesthetized, immobilized, artificially ventilated cats. Most suprageniculate nucleus cells were strongly sensitive to the direction of drifting gratings. The suprageniculate nucleus units had a clear preference for very low spatial frequencies with a mean of 0.05 cycle/deg. The spatial resolution was also very low with a mean of 0.16 cycle/deg. Most of the cells displayed low-pass spatial tuning characteristics, while the remainder of the units were band-pass tuned. The suprageniculate nucleus units were extremely narrowly tuned, to spatial frequencies with a mean spatial bandwidth of 1.07 octaves. A majority of the units responded optimally to high temporal frequencies, with a mean of 8.53 Hz. The temporal frequency tuning functions predominantly revealed a band-pass character, with a mean temporal bandwidth of 1.66 octaves. These results demonstrate that the neurons in the suprageniculate nucleus display particular spatial and temporal characteristics. The spatial and temporal tuning properties of the suprageniculate nucleus neurons are very similar to those of the superior colliculus and the anterior ectosylvian cortex, structures that provide the main visual afferentation toward the suprageniculate nucleus. This suggests their common function in motion perception, and especially in the recording of movements of the visual environment relative to the body, and the related behavioral action. PMID- 16359816 TI - Glutamate-induced Ca2+ influx in third-order neurons of salamander retina is regulated by the actin cytoskeleton. AB - Ligand-gated ion channels (ionotropic receptors) link to the cortical cytoskeleton via specialized scaffold proteins and thereby to appropriate signal transduction pathways in the cell. We studied the role of filamentous actin in the regulation of Ca influx through glutamate receptor-activated channels in third-order neurons of salamander retina. Staining by Alexa-Fluor 488-phalloidin, to visualize polymerized actin, we show localization of filamentous actin in neurites, and the membrane surrounding the cell soma. With Ca(2+) imaging we found that in dissociated neurons, depolymerization of filamentous actin by latrunculin A, or cytochalasin D significantly reduced glutamate-induced intracellular Ca(2+) accumulation to 53+/-7% of control value. Jasplakinolide, a stabilizer of filamentous actin, by itself slightly increased the glutamate induced Ca(2+) signal and completely attenuated the inhibitory effect when applied in combination with actin depolymerizing agents. These results indicate that in salamander retinal neurons the actin cytoskeleton regulates Ca(2+) influx through ionotropic glutamate receptor-activated channels, suggesting regulatory roles for filamentous actin in a number of Ca(2+)-dependent physiological and pathological processes. PMID- 16359817 TI - Enhanced Fos expression in glutamic acid decarboxylase immunoreactive neurons of the mouse periaqueductal grey during opioid withdrawal. AB - Previous studies using c-Fos immunohistochemistry suggest that a sub-population of neurons in the midbrain periaqueductal gray region is activated during opioid withdrawal. The neurochemical identity of these cells is unknown but cellular physiological studies have implicated GABAergic neurons. The present study investigated whether GABAergic neurons are activated in the mouse periaqueductal gray during opioid withdrawal using dual-antibody immunohistochemistry for Fos and glutamic acid decarboxylase. Both chronic opioid treatment and naloxone precipitated opioid withdrawal increased Fos expression in the periaqueductal gray, with the greatest increase being four-fold in the caudal ventrolateral subdivision following withdrawal. Neurons stained for both Fos and glutamic acid decarboxylase were greatly enhanced in all subdivisions of the periaqueductal gray following withdrawal, particularly in the lateral and ventrolateral divisions where the increase was up to 70-fold. These results suggest that activation of a subpopulation of GABAergic interneurons in the periaqueductal gray plays a role in opioid withdrawal. PMID- 16359818 TI - Increased and decreased activity elicits specific morphological adaptations of the neuromuscular junction. AB - The objective of this investigation was to compare the effects of decreased vs. increased activity on the neuromuscular system. Twenty-four young adult (7 weeks old) Sprague-Dawley rats were randomly assigned to three treatment groups (N=8/group). Increased activity was achieved by treadmill running for up to 1 h/day. Decreased activity was induced by muscle unloading via the hindlimb suspension model. Control animals engaged in normal weight bearing and ambulatory activity. At the end of the 10 week intervention period, animals were killed and soleus muscles were removed, quickly frozen, and examined using cytofluorescent (neuromuscular junctions) and histochemical (myofibers) procedures. Pre-synaptic morphology was quantified by measuring nerve terminal branching, and post synaptic assessment was conducted by staining acetylcholine receptors at the endplate. Myofiber profiles of solei were compiled by determining fiber size (cross-sectional area) and fiber type composition. Results show that exercise training significantly (P0.05). Co-incubation with 1% beta-CD or 1% HP-beta-CD exerted no significant effect on the P(app) of both propranolol and lucifer yellow (p>0.05), but co-incubation with 1.48% SBE-beta-CD lowered the P(app) of propranolol from (1.71+/-0.44)x10(-5) to (0.19+/-0.04)x10(-5)cm/s, which may be ascribed to the molecular complexation of propranolol with SBE-beta-CD. All three beta cyclodextrins exert no apparent impact on both (passive) transcellar and paracellular drug transports. PMID- 16359836 TI - Combination of adsorption by porous CaCO3 microparticles and encapsulation by polyelectrolyte multilayer films for sustained drug delivery. AB - Combination of adsorption by porous CaCO(3) microparticles and encapsulation by polyelectrolyte multilayers via the layer-by-layer (LbL) self-assembly was proposed for sustained drug release. Firstly, porous calcium carbonate microparticles with an average diameter of 5 microm were prepared for loading a model drug, ibuprofen (IBU). Adsorption of IBU into the pores was characterized by ultraviolet (UV), infrared (IR), thermogravimetric analysis (TGA), Brunauer Emmett-Teller (BET) experiment and X-ray diffraction (XRD). The adsorbed IBU amount Gamma was 45.1mg/g for one-time adsorption and increased with increasing adsorption times. Finally, multilayer films of protamine sulfate (PRO) and sodium poly(styrene sulfonate) (PSS) were formed on the IBU-loaded CaCO(3) microparticles by the layer-by-layer self-assembly. Amorphous IBU loaded in the pores of the CaCO(3) microparticles had a rapider release in the gastric fluid and a slower release in the intestinal fluid, compared with the bare IBU crystals. Polyelectrolyte multilayers assembled on the drug-loaded particles by the LbL reduced the release rate in both fluids. In this work, polymer/inorganic hybrid core-shell microcapsules were fabricated for controlled release of poorly water-soluble drugs. The porous inorganic particles are useful to load drugs in amorphous state and the polyelectrolyte multilayer films coated on the particle assuage the initial burst release. PMID- 16359837 TI - Antibacterial activity of Tabebuia impetiginosa Martius ex DC (Taheebo) against Helicobacter pylori. AB - The growth-inhibiting activity of Tabebuia impetiginosa Martius ex DC dried inner bark-derived constituents against Helicobacter pylori ATCC 43504 was examined using paper disc diffusion and minimum inhibitory concentration (MIC) bioassays. The activity of the isolated compounds was compared to that of the commercially available anti-Helicobacter pylori agents, amoxicillin, metronidazole, and tetracycline. The biologically active components of Tabebuia impetiginosa dried inner bark (taheebo) were characterized by spectroscopic analysis as 2 (hydroxymethyl)anthraquinone, anthraquinone-2-carboxylic acid, and 2-hydroxy-3-(3 methyl-2-butenyl)-1,4-naphthoquinone (lapachol). With the paper disc diffusion assay 2-(hydroxymethyl)anthraquinone exhibited strong activity against Helicobacter pylori ATCC 43504 at 0.01 mg/disc. Anthraquinone-2-carboxylic acid, lapachol and metronidazole were less effective, exhibiting moderate anti Helicobacter pylori activity at 0.1 mg/disc. Amoxicillin and tetracycline were the most potent compounds tested, displaying very strong activity at 0.005 mg/disc. 2-(Hydroxymethyl)anthraquinone exhibited moderate activity at this dose. Tetracycline still had strong activity at 0.001 mg/disc while amoxicillin had little activity at this dose. In the MIC bioassay, 2-(hydroxymethyl)anthraquinone (2 microg/mL), anthraquinone-2-carboxylic acid (8 microg/mL), and lapachol (4 microg/mL) were more active than metronidazole (32 microg/mL) but less effective than amoxicillin (0.063 microg/mL) and tetracycline (0.5 microg/mL). The anti Helicobacter pylori activity of seven 1,4-naphthoquinone derivatives (structurally related to lapachol), 1,4-naphthoquinone, 5,8-dihydroxy-1,4 naphthoquinone (naphthazarin), 2-methyl-1,4-naphthoquinone (menadione), 2-hydroxy 1,4-naphthoquinone (lawsone), 5-hydroxy-2-methyl-1,4-naphthoquinone (plumbagin), 5-hydroxy-1,4-naphthoquinone (juglone), and 2,3-dichloro-1,4-naphthoquinone (dichlone) was also evaluated using the paper disc assay. Menadione and plumbagin were the most potent compounds tested with the later still exhibiting very strong activity at 0.001 mg/disc. Menadione, juglone and tetracycline had strong activity at this low dose while the latter two compounds and amoxicillin had very strong activity at 0.005 mg/disc. Lawsone was unusual in that it had very strong activity at 0.1 and 0.05 mg/disc but weak activity at doses of 0.01 mg/disc and lower. Naphthazalin, lapachol and dichlone had similar activities while metronidazole had the lowest activity of all compounds tested. These results may be an indication of at least one of the pharmacological actions of taheebo. The Tabebuia impetiginosa dried inner bark-derived materials, particularly 2 (hydroxymethyl)anthraquinone, merit further study as potential Helicobacter pylori eradicating agents or lead compounds. PMID- 16359838 TI - Protective effect of Phyllanthus fraternus against allyl alcohol-induced oxidative stress in liver mitochondria. AB - The effect of administration of allyl alcohol on the oxidative stress and the protective effect due to administration of an aqueous extract of Phyllanthus fraternus against allyl alcohol-induced damage in liver mitochondria were studied. When rats were treated with allyl alcohol, the rate of mitochondrial respiration was decreased significantly with both NAD(+)- and FAD-linked substrates. The respiratory control ratio, an index of membrane integrity and the P/O ratio, a measure of phosphorylation efficiency also decreased significantly. There was a significant increase in the lipid peroxide level and the protein carbonyl content. A significant decrease was observed in the total sulphydryl groups and a significant increase in the generation of superoxide radicals. Administration of rats with an aqueous extract of Phyllanthus fraternus (100 mg/kg) prior to allyl alcohol administration showed protection of 72, 40 and 80% using glutamate+malate (NADH oxidation) and 77, 54 and 20% using succinate as substrate on state 3, RCR and P/O ratio, respectively. The protection on lipid peroxide level was 88 and 91% in homogenate and mitochondria, respectively. In case of protein carbonyls, total sulphydryl groups and on the generation of superoxide radicals the protection was 99, 59 and 53%, respectively. PMID- 16359839 TI - Estrogenic and antioxidant effects of a phytoestrogen daidzein on ovarian germ cells in embryonic chickens. AB - The estrogenic and antioxidant effects of the phytoestrogen daidzein (DAI) on germ cell proliferation were evaluated by a chicken ovarian germ-somatic cell coculture model. Ovarian cells were dispersed from 18-day-old embryos, cultured in serum-free McCoy's 5A medium and challenged with DAI alone or in combinations with estrogen receptor antagonist tamoxifen for 48 h. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). The labeling index (LI) was determined for germ cells. Results showed that DAI significantly increased the number of germ cells (P<0.05) and this stimulating effect was inhibited by tamoxifen in a dose-dependent manner. Furthermore, PCNA-LI of germ cells displayed similar changes with the number of germ cells. To estimate the antioxidant action of DAI, ovarian cells were exposed to the reactive oxygen species (ROS)-producing system hypoxanthine/xanthine oxidase (HX/XO). The changes of superoxide dismutase (SOD) activity and glutathione (GSH) level were measured for estimation of the antioxidant status. Ovarian cells were severely damaged by free radicals and this deteriorating effect could be prevented by DAI. Moreover, HX/XO-induced decrease in SOD activity and GSH level was restored by DAI (P<0.05). These results indicated that DAI promoted proliferation of cultured ovarian germ cells by estrogenic action and attenuated ROS-induced toxicity by antioxidant action in embryonic chickens. PMID- 16359840 TI - In vitro methods in human drug biotransformation research: implications for cancer chemotherapy. AB - Anticancer drugs have a complex pharmacological and toxicological profile with a narrow therapeutic index. It is therefore critical to understand the factors that contribute to the marked intersubject variability in the pharmacokinetics and pharmacodynamics often observed with anticancer compounds. Since hepatic and extra-hepatic drug metabolism represents a major drug disposition pathway, extensive efforts are made to thoroughly investigate metabolism of anticancer compounds during the pre-clinical and clinical development phases as well as to address issues encountered during the clinical use of an approved drug. In recent years there has been a significant paradigm shift in pre-clinical/non-clinical drug metabolism studies. Most importantly, this has included a reduced reliance on animal models and increased use of human tissues (i.e. human liver microsomes and other cellular fractions, primary culture of human hepatocytes, cDNA expressed human-specific enzymes and cell-based reporter assays). Typically, experiments are performed using these tools to identify the phase I and/or phase II enzymes involved in metabolism of the drug/investigational agent and for metabolic fingerprinting. Additionally, issues pertaining to the rate, extent and mechanism(s) of the inhibition or induction of the metabolic pathways are also investigated. These studies provide important clues about various aspects of the disposition of a therapeutic agent including first-pass metabolism, elimination half-life, overall bioavailability and the potential for drug-drug interactions. The methodologies used for in vitro assessment of drug metabolism and their applications to drug development and clinical therapeutics with special emphasis on anticancer drugs are reviewed in this manuscript. PMID- 16359841 TI - Capucin: a novel striatal marker down-regulated in rodent models of Huntington disease. AB - In an initial study, we compared quantitative transcriptome data across mouse brain territories using the serial analysis of gene expression method. Among the novel regional markers that we discovered, we focused on a striatum-enriched transcript with no available experimental cDNA sequence. Here, we report its cloning, gene structure, and detailed distribution in mouse brain. Quantitative RT-PCR and in situ hybridization demonstrated predominant expression in dorsolateral striatum. We therefore named it capucin for caudate-and putamen enriched sequence. Mouse capucin is a 237-amino-acid protein, without any registered ortholog in mammalian species. It contains no recognizable motif other than two predicted carboxy-terminal transmembrane domains. When expressed in fusion with a fluorescent protein, it localized to the Golgi apparatus in two mammalian cell lines. Interestingly, we observed a significant down-regulation of capucin mRNA levels in two rodent models of Huntington disease, indicating a possible contribution to the pathogenesis of this disorder. PMID- 16359842 TI - Spontaneous eye-blinking and stereotyped behavior in older persons with mental retardation. AB - Previous research indicates that abnormal stereotyped movements are associated with central dopamine dysfunction and that eye-blink rate is a noninvasive, in vivo measure of dopamine function. We measured the spontaneous eye-blinking and stereotyped behavior of older adults with severe/profound mental retardation living in a state mental retardation facility. Analyses revealed that the mean eye-blink rate of the residents that engaged in stereotypy was significantly lower than the rate for residents who did not exhibit stereotypy. Moreover, the stereotypy group also demonstrated greater variability in interblink intervals. These results provide further empirical support for the involvement of dopamine in stereotyped behavior and are consistent with an emerging motor control model of stereotypy. PMID- 16359844 TI - A neural network model of Parkinson's disease bradykinesia. AB - Parkinson's disease (PD) is caused by dopamine (DA) depletion consequent to cell degeneration in the substantia nigra pars compacta (SNc) and the ventral tegmental area (VTA). Although computational analyses of PD have focused on DA depletion in DA-recipient parts of the basal ganglia, there is also extensive DAergic innervation of the frontal and parietal cortex as well as the spinal cord. To understand PD bradykinesia, a comprehensive network model is needed to study how patterns of DA depletion at key cellular sites in the basal ganglia, cortex and spinal cord contribute to disordered neuronal and spinal cord activity and other PD symptoms. We extend a basal ganglia-cortico-spinal circuit for control of voluntary arm movements by incorporating DAergic innervation of cells in the cortical and spinal components of the circuit. The resultant model simulates successfully several of the main reported effects of DA depletion on neuronal, electromyographic (EMG), and movement parameters of PD bradykinesia. PMID- 16359845 TI - Integron-associated imipenem resistance in Acinetobacter baumannii isolated from a regional hospital in Taiwan. AB - We investigated the genetic properties of imipenem-resistant Acinetobacter baumannii collected from a regional hospital in Taiwan. Pulsed-field gel electrophoresis demonstrated that the isolates were genetically diverse. Polymerase chain reaction, DNA sequencing, and DNA-DNA hybridisation showed that the bla(IMP-1) gene resided as a cassette in a plasmid-borne class 1 integron in two isolates. The majority of the resistant isolates were plasmid-less and carried no bla(IMP), bla(VIM) or bla(CFI) genes, indicating that other uncharacterised metallo-beta-lactamases or mechanisms other than enzyme production are involved in carbapenem resistance in this group of A. baumannii. We conclude that multidrug resistance of A. baumannii was a combined effect of lateral gene transfer and clonal spread of multiple resistant clones. Strict measures should be implemented to control the further spread of resistance. PMID- 16359846 TI - High-level expression of IMP-5 carbapenemase owing to point mutation in the -35 promoter region of class 1 integron among Pseudomonas aeruginosa clinical isolates. AB - Eight strains of Pseudomonas aeruginosa producing IMP-5 carbapenemases were collected from three Portuguese hospitals. All isolates were epidemiologically unrelated. The bla(IMP-5) gene was inserted into a class 1 integron previously reported in Acinetobacter baumannii 65FFC. Expression of the the bla(IMP-5) gene in P. aeruginosa has been shown to be driven by the P(1) promoter [TTGATA] in which the cytosine was replaced by thymine, which caused an increase in transcription of bla(IMP-5) that was confirmed by site-directed mutagenesis. The minimum inhibitory concentrations (MICs) of imipenem for Escherichia coli pGMLA-1 (the recombinant from A. baumannii 65FFC) and E. coli pATG-2 (the recombinant from P. aeruginosa isolates) were 0.5 mg/L and >32 mg/L, respectively. This study reports the spread of a class 1 integron In76 with a new point mutation in the P(1) promoter sequence leading to overproduction of IMP-5. PMID- 16359847 TI - Simultaneous quantitation of five active principles in a pharmaceutical preparation: development and validation of a near infrared spectroscopic method. AB - A near infrared spectroscopic method for the simultaneous determination of the active principles paracetamol, ascorbic acid, dextrometorphan hydrobromide, caffeine and chlorpheniramine maleate in a pharmaceutical preparation was developed. The five active principles are quantified using a partial least squares regression method (PLS1). The proposed method is applicable over a wide analyte concentration range (0.04-6.50 wt.%), so it requires careful selection of the calibration set. Also, there is the difficulty of ensuring thorough homogenization of the product. The method was validated in accordance with the ICH standard and the EMEA validation guidelines for NIR spectroscopy by determining its selectivity, linearity, accuracy, precision and robustness. Based on the results, it is an effective alternative to existing choices (HPLC and redox titrimetry) for the same purpose. PMID- 16359848 TI - Development and evaluation of an in vitro method for prediction of human drug absorption I. Assessment of artificial membrane composition. AB - A study was undertaken aimed at developing a dynamic in vitro method based on the use of artificial membranes for screening and prediction of the absorption properties of drugs. The development of a suitable artificial membrane simulating the behaviour of the natural gastrointestinal one is the critical step for the fulfillment of this research and it has been the aim of the present work. A series of filters of different chemical nature and different characteristics (pore size, percent of porosity, thickness) were evaluated and compared in order to select the most suitable one to use as support for the artificial lipid membrane preparation. Permeation experiments were performed by using naproxen as reference model drug. The target was to achieve a naproxen apparent permeability (P(app)) value close to that previously obtained using a Caco-2 cell dynamic methodology (4.88 x 10(-5)cms(-1)), which related well to the fraction of dose absorbed in vivo in human. Filters of cellulose nitrate-acetate mixture with a porosity of 70% and 0.025 microm pore size were selected on the basis of the highest reproducibility of results and suitable drug P(app) value obtained. Mixture design experiments were then carried out in order to optimize the composition of the lipid mixture to use for porous support impregnation to obtain the desired P(app) value. Sixteen different lipid mixtures were prepared according to mixture experimental plan and evaluated by diffusion experiments. The contour plot obtained was used to select an optimum lipidic mixture (cholesterol 2.10%; Lipoid((R)) E 80 1.70%; n-octanol 96.2%) for the cellulose nitrate-acetate membrane impregnation. The experimental response obtained from diffusion experiments performed by utilising the optimised lipidic impregnation mixture, was 4.88 x 10(-5)cms(-1), matching exactly the prefixed target. PMID- 16359849 TI - [Superior mesenteric artery syndrome: a cause of vomiting in children. Report of 3 cases]. AB - Duodenal obstruction by Superior Mesenteric Artery (SMA) is a misdiagnosed vomiting syndrome in children. Several factors are involved, including rapid weight loss, rapid statural growth without weight augmentation. Diagnosis is suspected when an improvement is achieved by ventral decubitus and it is confirmed by plain films of the abdomen, GI study with barium and echography, measuring the aortomesenteric angle (inferior to 25-30 degrees ). Patients must at first be treated conservatively. Surgery is indicated for occlusive episodes with unsuccessful conservative therapy. The authors report 3 cases with different clinical presentation. However, all the patients presented important weight loss and vomiting. PMID- 16359850 TI - [The pediatric pathology: a speciality with a future?]. PMID- 16359851 TI - Automatic identification of confusable drug names. AB - OBJECTIVE: Many hundreds of drugs have names that either look or sound so much alike that doctors, nurses and pharmacists can get them confused, dispensing the wrong one in errors that can injure or even kill patients. METHODS AND MATERIAL: We propose to address the problem through the application of two new methods-one based on orthographic similarity ("look-alike"), and the other based on phonetic similarity ("sound-alike"). In order to compare the effectiveness of the new methods for identifying confusable drug names with other known similarity measures, we developed a novel evaluation methodology. RESULTS: We show that the new orthographic measure (BI-SIM) outperforms other commonly used measures of similarity on a set containing both look-alike and sound-alike pairs, and that a new feature-based phonetic approach (ALINE) outperforms orthographic approaches on a test set containing solely sound-alike pairs. However, an approach that combines several different measures achieves the best results on two test sets. CONCLUSION: Our system is currently used as the basis of a system developed for the U.S. Food and Drug Administration for detection of confusable drug names. PMID- 16359852 TI - Epinephrine quantification in pharmaceutical formulations utilizing plant tissue biosensors. AB - A plant tissue biosensor associated with flow injection analysis is proposed to determine epinephrine in pharmaceutical samples. The polyphenol oxidase enzymes present in the fibers of a palm tree fruits (Livistona chinensis), catalyses the oxidation of epinephrine to epinephrinequinone as a primary product. This product is then electrochemically reduced (at -0.10 V versus Ag/AgClsat) on the biosensor surface and the resulting current is used for the quantification of epinephrine. The biosensor provides a linear response for epinephrine in the concentration range from 5.0 x 10(-5) to 3.5 x 10(-4) mol l(-1). The limit of detection estimated for this interval was 1.5 x 10(-5) mol l(-1) and the correlation coefficient of 0.998, working under a flow rate of 2.0 ml min(-1) and using a sample loop of 100 microl. The repeatability (R.S.D. for 10 consecutive determinations of a 3.0 x 10(-4) mol l(-1) epinephrine solution) was 3.1%. The results obtained by the method here proposed were compared with the official UV spectrophotometric procedure and also using a plant tissue reactor. The responses obtained with the proposed strategies were in good agreement with both ways of analyses, whereas the values obtained by the official spectrophotometric method was strongly affected by benzoic acid, present in the formulation of pharmaceutical product utilized for inhalation. Such favorable results obtained with the carbon paste biosensor or utilizing the bioreactor, joined with the simplicity of its preparation turns these procedures very attractive for epinephrine quantification in pharmaceutical products. PMID- 16359853 TI - Controlling bacterial physiology for optimal expression of gene reporter constructs. AB - Bacterial biosensors for the detection of pollutants are based on the regulatory elements that control the corresponding degradation pathways. An increasing number of catabolic pathways under the control of specific regulators are now known to be influenced by the presence of alternative carbon sources, which to different extents repress expression of the pathway despite the presence of the inducer. The molecular basis underlying the control of each catabolic pathway is different, although all sense a high energy state of the cell resulting from the presence of more favourable carbon sources. Biosensor tests mimicking field conditions point to global regulation being relevant for biosensor performance; thus, this global regulation must be taken into account when designing whole-cell biosensors. PMID- 16359854 TI - Transcriptional regulators a la carte: engineering new effector specificities in bacterial regulatory proteins. AB - For many regulators of bacterial biodegradation pathways, small molecule/effector binding is the signal for triggering transcriptional activation. Thus, regulation results from a cross-talk between chemicals sensed by transcriptional factors and operon expression status. These features can be utilised in the construction of biosensors for a wide range of target compounds as, in principle, any regulatory protein whose activity is modulated by binding to a small molecule can have its effector/inducer profile artificially altered. The cognate specificities of a number of regulatory proteins have been modified as an astute approach to developing, among others, bacterial biosensors for environmentally relevant compounds. PMID- 16359855 TI - mTOR and cancer: reason for dancing at the crossroads? AB - Recent successes using Gleevec for the treatment of chronic myelogenous leukemia and gastrointestinal stromal tumors have provided proof that strategies to target signal transduction pathways mutated in human cancers can work. However, the application of this strategy to other cancers has been slow. Central to alleviating this impedance is the molecular characterization of the tumors. There is an urgent need to translate basic scientific findings into relevant, clinically applicable molecular diagnostic assays. PMID- 16359856 TI - p63: oncogene or tumor suppressor? AB - p53, the original member of the family of genes now known to include p63 and p73, was first heralded as an oncogene because of its potent transformation capabilities and its robust expression in human tumors. However, it was later discovered that only mutant p53 was oncogenic, and that wild type p53 functioned as a tumor suppressor. Decades later, p63, the newest member of this gene family, is involved in a similar controversy: is p63 an oncogene or a tumor suppressor? Recent progress on understanding the in vivo role of p63 in cancer has focused primarily on investigating its involvement in the tumor-suppressive mechanism of apoptosis, by analyzing mouse models to assess its tumor-suppressive capabilities, and by assessing its expression in human cancers. PMID- 16359857 TI - Drosophila models for cancer research. AB - Drosophila is a model system for cancer research. Investigation with fruit flies has facilitated a number of important recent discoveries in the field: the hippo signaling pathway, which coordinates cell proliferation and death to achieve normal tissue size; 'social' behaviors of cells, including cell competition and apoptosis-induced compensatory proliferation, that help ensure normal tissue size; and a growing understanding of how oncogenes and tumor suppressors cooperate to achieve tumor growth and metastasis in situ. In the future, Drosophila models can be extended beyond basic research in the search for human therapeutics. PMID- 16359858 TI - Chemical genetic approaches to the development of cancer therapeutics. AB - Dysregulation of kinase-based signal transduction networks contributes to multiple aspects of malignancy. Chemical genetic approaches interrogate perturbed signaling in the immediate context of small molecule inhibitor treatment. In recent years, such approaches have identified new kinase targets, clarified the impact of poly-specific inhibition using agents for which at least one primary target is known, and have identified targets for which combinatorial inhibition leads to improved efficacy. Elucidation of the mechanisms through which specific small molecule drug-like agents impact crucial cancer pathways should yield important and clinically translatable insights into the use of similar agents in patients. PMID- 16359859 TI - Methylenetetrahydrofolate reductase 677C>T polymorphism and gastric cancer susceptibility in Mexico. AB - This study investigated whether methylenetetrahydrofolate reductase MTHFR 677C>T polymorphism modified gastric cancer (GC) risk independently as well as in combination with folate intake and alcohol consumption. A hospital-based case control study of 201 cases and 427 controls was conducted in three geographical areas of Mexico, between 1994 and 1996. The MTHFR 677T allele frequency was 51.0% in cases compared with 45.3% in controls. After controlling for dietary sources of folate, alcohol intake and other selected variables, a significant increase in GC risk was found among carriers of the 677TT genotype compared with those with the 677CC genotype (odds ratio (OR) 1.62, 95% confidence interval (CI) 1.00 2.59), with a significant trend (P = 0.048). There were no significant interactions between the MTHFR polymorphism and consumption of folate and alcohol. Our results suggest that the high prevalence of MTHFR 677T allele may be a contributor to the high rate of morbidity and mortality in GC in Mexico. PMID- 16359860 TI - Cancer survival in the elderly: effects of socio-economic factors and health care system features (ELDCARE project). AB - The purpose of the ELDCARE project is to study differences in cancer survival for elderly patients by country, taking into account the socio-economic conditions and the characteristics of health care systems at the ecological level. Fifty three European cancer registries, from 19 countries, participating in the EUROCARE 3 programme, collected information to compute relative survival on patients aged 65-84 years, diagnosed over the period 1990-1994. National statistics offices provided the macro-economic and labour force indicators (gross domestic product, total health expenditure, and proportion of people employed in the agriculture sector) as well as the features of national health care systems. Survival for several of the cancer sites had high positive Pearson's correlations (r) with the affluence indicators (usually r>0.7), but survival for the poor prognosis cancers (lung, ovary, stomach) and for cervix uteri was not so well correlated. Among the medical resources considered, the number of computed tomography scanners was the variable most related to survival in the elderly; the number of total health practitioners in the country did not show any relationship. Survival was related to the marital status of elderly women more strongly than for men and younger people. The highest correlations of survival with the percentage of married elderly women in the population were for cancers of the rectum (r=0.79) and breast (r=0.66), while survival correlated negatively with the proportion of widows for most cancers. Being married or widowed is for elderly people, in particular elderly women, an important factor influencing psychological status, life habits and social relationships. Social conditions could play a major role in determining health outcomes, particularly in the elderly, by affecting access to health care and delay in diagnosis. PMID- 16359861 TI - Amount of leachant and water absorption levels of wood treated with borates and water repellents. AB - Wood protection efficacy of borates against biological agents, flame retardancy, and suitability to the environment is well known. Since borates can be applied to timber as water based solutions, they are preferred economically as well. Even though they are highly mobile in wood, boron compounds are widely used in timber preservation. Borates migrate in liquid and increase the hygroscopicity of wood in damp conditions. This study deals with the physical restriction of water access in wood by impregnating water repellent agents into wood to limit amount of leachant and water absorption levels of wood after boron treatment. Borates were incorporated with polyethylene glycol-400 (PEG-400) their bulking effect in wood was considered. Results indicated that the amount of leachates from wood treated with borates in PEG-400 was remarkably higher compared to those of wood treated with the aqueous solutions of borates. Water absorption (WA) levels of wood treated with aqueous solutions of borates were higher than those of their treated samples with the solutions in PEG-400. Secondary treatments of wood with the water repellent (WR) chemicals following borate impregnation reduced the leaching of chemicals from wood in water and also WA of the specimens were less than those of the wood treated with only borates from aqueous and PEG solutions. Styrene (St) was the most effective monomer among the other agents used in terms of immobility effect on borates and WA. PMID- 16359862 TI - Optimization of alkali-catalyzed transesterification of Pongamia pinnata oil for production of biodiesel. AB - Studies were carried out on transesterification of Karanja oil with methanol for the production of biodiesel. The reaction parameters such as catalyst concentration, alcohol/oil molar ratio, temperature, and rate of mixing were optimized for production of Karanja oil methyl ester (KOME). The fatty acid methyl esters content in the reaction mixture were quantified by HPLC and 1H NMR method. The yield of methyl esters from Karanja oil under the optimal condition was 97-98%. PMID- 16359863 TI - Synthesis and antimycobacterial activity of some alkyl [5-(nitroaryl)-1,3,4 thiadiazol-2-ylthio]propionates. AB - Two series of 2- and 3-[5-(nitroaryl)-1,3,4-thiadiazol-2-ylthio, sulfinyl and sulfonyl] propionic acid alkyl esters were synthesized and screened for antituberculosis activity against Mycobacterium tuberculosis H37Rv using the BACTEC 460 radiometric system. The MIC values for the compounds showing more than 90% inhibition were determined. The result of comparison between two groups of data exhibited that among the synthesized derivatives, the compound propyl 3-[5 (5-nitrothiophen-2-yl)-1,3,4-thiadiazol-2-ylthio]propionate was the most active one (MIC=1.56 microgml(-1)). PMID- 16359864 TI - Synthesis and antihyperglycemic evaluation of various protoberberine derivatives. AB - Various berberine derivatives (2-17) were synthesized and their antihyperglycemic activities were evaluated in a model of beta-cell-membrane chromatography and a model of alloxan-induced diabetes mice. The results indicated that compounds 5 and 14 exhibited antihyperglycemic activity. Their structure-activity relationships were discussed. PMID- 16359865 TI - Synthesis and structure-activity relationships of novel pyrrolocarbazole lactam analogs as potent and cell-permeable inhibitors of poly(ADP-ribose)polymerase-1 (PARP-1). AB - A series of novel pyrrolocarbazole lactams was identified as potent PARP-1 inhibitors in vitro and in a PC12 cellular NAD(+) depletion assay. The SAR trends of substituents at the 3-position, as well as the effect of blocking the indole or lactam NH-groups of the template by methylation or formylation, are discussed in relation to molecular modeling studies. PMID- 16359866 TI - Partial structures of ketoconazole as modulators of the large conductance calcium activated potassium channel (BK(Ca)). AB - A series of partial structures of ketoconazole has been synthesized and tested for activity on the large conductance calcium-activated potassium channel (BK) in bovine smooth muscle cells. This has provided openers and blockers of the channel. The results suggest that the phenyl and phenoxy moieties are important for interaction with BK, whereas the imidazole group is unimportant. The properties of the phenoxy moiety seem to determine whether the compounds act to open or block the channel. PMID- 16359868 TI - Stepping over obstacles of different heights: kinematic and kinetic strategies of leading limb in hemiplegic children. AB - Twelve hemiplegic children (HC) and five normally developing children (NDC) were instructed to walk barefoot over a level surface and to cross obstacles of various heights (0%, 10%, 15% and 20% of their leg lengths). Kinematic and kinetic strategies of their leading limb were mainly examined. For the starting position of the trials HC was adjusted so that the affected limb was the leading one, while NDC step over the obstacle with their preferred selection of leading and trailing limb. The results demonstrated that the crossing speed was decreased and the crossing swing phase was increased in both groups. The toe vertical clearance augmented with the presence of the obstacle, but it remained constant with obstacle heights. The toe and heel horizontal distances of the leading limb remained constant with all obstacle heights. The toe horizontal distances of the trailing limb in HC increased with obstacle heights outlying a peculiar strategy. A comparison of joint angles and power variations revealed substantial differences in height changes between the two groups: HC exhibited a preferential modulation of pelvis and hip joints and a reduced modulation of knee and ankle joints. The knee flexor limitation in HC was accomplished by knee power variations near the toe-off. HC showed also an increased ankle involvement of the support limb, i.e. the almost normal limb. Collectively, the results provided insights into multijoint variations of locomotor act caused by visible obstacle in HC and indicate that a correct rehabilitation treatment should be focused on the use of dynamic constraints preferentially at the plegic knee. PMID- 16359869 TI - Charting a course with TiBS. PMID- 16359870 TI - Direct PCR amplification of various modified DNAs having amino acids: convenient preparation of DNA libraries with high-potential activities for in vitro selection. AB - We synthesized modified 2'-deoxyuridine triphosphates bearing amino acids at the C5 position and investigated their substrate properties for KOD Dash DNA polymerase during polymerase chain reaction (PCR). PCR using C5-modified dUTP having an amino acyl group (arginyl, histidyl, lysyl, phenylalanyl, tryptophanyl, leucyl, prolyl, glutaminyl, seryl, O-benzyl seryl or threonyl group) gave the corresponding full-length PCR products in good yield. Although dUTP analogues bearing aspartyl, glutamyl or cysteinyl were found to be poor substrates for PCR catalyzed by KOD Dash DNA polymerase, optimization of the reaction conditions resulted in substantial generation of full-length product. In the case of reaction using dUTP analogue having a cysteinyl group, addition of a reducing agent improved the reaction yield. Thus, PCRs using KOD Dash DNA polymerase together with amino acyl dUTP provide convenient and efficient preparation of various modified DNA libraries with potential protein-like activities. PMID- 16359871 TI - Genetic vaccines protect red seabream, Pagrus major, upon challenge with red seabream iridovirus (RSIV). AB - We have investigated the protective effect of immunization of juvenile red seabream, Pagrus major, with DNA plasmids encoding the viral major capsid protein (MCP) and an open reading frame (ORF) containing a transmembrane domain against red seabream iridovirus (RSIV). The expression of the MHC class I transcript in the DNA-vaccinated fish was significantly upregulated at the 15th day post vaccination and the relative level of expression was maintained until the 30th day post-vaccination. This pattern of expression was similar in fish vaccinated with a commercially prepared formalin-inactivated RSIV vaccine. In vaccine efficiency tests, the relative percentage survival (RPS) of fish receiving the DNA vaccines and their combination ranged from 42.8 to 71.4% in two experimental runs, and these were significantly different from the control groups. Our results clearly demonstrate that DNA vaccines are able to induce robust protection in fish against RSIV infection, and a cellular immune response as shown by the upregulation of the MHC class I transcript after vaccination, which may be associated with such protection. PMID- 16359872 TI - Further evaluation of an EMG technique for assessment of the deep cervical flexor muscles. AB - A novel surface electromyographic (EMG) technique was recently described for the detection of deep cervical flexor muscle activity. Further investigation of this technique is warranted to ensure EMG activity from neighbouring muscles is not markedly influencing the signals recorded. This study compared deep cervical flexor (DCF) muscle activity with the activity of surrounding neck and jaw muscles during various anatomical movements of the neck and jaw in 10 volunteer subjects. DCF EMG activity was recorded with custom electrodes inserted via the nose and fixed by suction to the posterior mucosa of the oropharynx. Surface electrodes were placed over the sternocleidomastoid, anterior scalene, masseter and suprahyoid muscles. Positioned in supine, subjects performed isometric cranio cervical flexion, cervical flexion, right and left cervical rotation, jaw clench and resisted jaw opening. Across all movements examined, EMG amplitude of the DCF muscles was greatest during neck movements that would require activity of the DCF muscles, particularly during cranio-cervical flexion, their primary anatomical action. The actions of jaw clench and resisted jaw opening demonstrated significantly less DCF EMG activity than the cranio-cervical flexion action (p<0.05). Across all other movements, the neighbouring neck and jaw muscles demonstrated greatest EMG amplitude during their respective primary anatomical actions, which occurred in the absence of increased EMG amplitude recorded from the DCF muscles. The finding of substantial EMG activity of the DCF muscles only during neck actions that would require their activity, particularly cranio cervical flexion, and not during actions involving the jaw, provide further assurance that the majority of myoelectric signals detected from the nasopharyngeal electrode are from the DCF muscles. PMID- 16359873 TI - Perception of matching and conflicting audiovisual speech in dyslexic and fluent readers: an fMRI study at 3 T. AB - We presented phonetically matching and conflicting audiovisual vowels to 10 dyslexic and 10 fluent-reading young adults during "clustered volume acquisition" functional magnetic resonance imaging (fMRI) at 3 T. We further assessed co variation between the dyslexic readers' phonological processing abilities, as indexed by neuropsychological test scores, and BOLD signal change within the visual cortex, auditory cortex, and Broca's area. Both dyslexic and fluent readers showed increased activation during observation of phonetically conflicting compared to matching vowels within the classical motor speech regions (Broca's area and the left premotor cortex), this activation difference being more extensive and bilateral in the dyslexic group. The between-group activation difference in conflicting > matching contrast reached significance in the motor speech regions and in the left inferior parietal lobule, with dyslexic readers exhibiting stronger activation than fluent readers. The dyslexic readers' BOLD signal change co-varied with their phonological processing abilities within the visual cortex and Broca's area, and to a lesser extent within the auditory cortex. We suggest these findings as reflecting dyslexic readers' greater use of motor-articulatory and visual strategies during phonetic processing of audiovisual speech, possibly to compensate for their difficulties in auditory speech perception. PMID- 16359874 TI - Decomposition of metabolic brain clusters in the frontal variant of frontotemporal dementia. AB - Previous studies that measured brain activity in frontotemporal dementia (FTD) used univariate analyses, examining each region of interest separately. We explored in a multicenter European research program the principal brain clusters characterized by a common variability in cerebral metabolism in FTD. Seventy patients with frontal variant (fv) FTD were selected according to international clinical recommendations; principal component analysis (PCA) was performed on FDG PET metabolic images, looking for covariance clusters in this large population. A first metabolic cluster included most of the lateral and medial prefrontal cortex, bilaterally; PC1 scores correlated with performances on memory and executive neuropsychological tasks. Moreover, FDG-PET images in fv-FTD were further characterized by a metabolic covariance in two clusters comprising the subcallosal medial frontal region, the temporal pole, medial temporal structures and the striatum, separately in the left and in the right hemisphere. The study provides original data-driven arguments for metabolic involvement of separate brain clusters in the rostral limbic system, corresponding to pathological poles differentially affected in each FTD patient. PMID- 16359875 TI - The complete mitochondrial genomes of the sea lily Gymnocrinus richeri and the feather star Phanogenia gracilis: signature nucleotide bias and unique nad4L gene rearrangement within crinoids. AB - Complete DNA sequences have been determined for the mitochondrial genomes of the crinoids Phanogenia gracilis (15892 bp) and Gymnocrinus richeri (15966 bp). The mitochondrial genetic map of the stalkless feather star P. gracilis is identical to that of the comatulid feather star Florometra serratissima (Scouras, A., Smith, M.J., 2001. Mol. Biol. Evol. 18, 61-73). The mitochondrial gene order of the stalked crinoid G. richeri differs from that of F. serratissima and P. gracilis by the transposition of the nad4L protein gene. The G. richeri nad4L mitochondrial map position is unique among metazoa and is likely a derived feature in this stalked crinoid. Nucleotide compositional analyses of protein genes encoded on the major sense strand confirm earlier conclusions regarding a crinoid-distinctive T over C bias. All three crinoids exhibit high T levels in third codon positions, whereas other echinoderm classes favor A or C in the third codon position. The nucleotide bias is reflected in the relative synonymous codon usage patterns of crinoids versus other echinoderms. We suggest that the nucleotide bias of crinoids, in comparison to other echinoderms, indicates that a physical inversion of the origin of replication has occurred in the crinoid lineage. Evolutionary rate tests support the use of the cytochrome b (cob) gene in molecular phylogenetic analyses of echinoderms. A consensus echinoderm tree was generated based on cytochrome b nucleotide alignments that placed the asteroids as a sister group to a clade containing the ophiuroids and the (echinoids+holothuroids) with the crinoids basal to the rest of the echinoderm classes: [Crinoid,(Asteroid,(Ophiuroid,(Echinoid,Holothuroid)))]. PMID- 16359876 TI - Effects of helium-neon laser on the mucopolysaccharide induction in experimental osteoarthritic cartilage. AB - OBJECTIVE: To investigate the effects of mucopolysaccharide induction after treatment by low power laser for experimental osteoarthritis (OA). METHODS: Seventy-two rats with three different degrees of papain induced OA over right knee joints were collected for helium-neon (He-Ne) laser treatment. The severity of induced arthritis was measured by 99mTc bone scan and classified into three groups (I-III) by their radioactivity ratios (right to left knee joints). The rats in each group were further divided into study subgroups (Is, IIs, and IIIs) and control subgroups (Ic, IIc, and IIIc) randomly. The arthritic knees in study subgroups received He-Ne laser treatment, and those in controls received sham laser treatment. The changes of arthritic severity after treatment and follow-up 2 months later were measured. The histopathological changes were evaluated through light microscope after disarticulation of sections (H.E. stain), and the changes of mucopolysaccharide density in cartilage matrix were measured by Optimas scanner analyzer after Alcian blue (AB) stain. The densities of mucopolysaccharide induced after treatment in arthritic cartilage were compared and correlated with their histopathological changes. RESULTS: The density of mucopolysaccharide rose at the initial stage of induced arthritis, and decreased progressively in later stages. The densities of mucopolysaccharide in treated rats increased upon complete laser treatment more than those of the controls, which is closely related with the improvement in histopathological findings, but conversely with the changes in arthritic severity. CONCLUSION: He-Ne laser treatment will enhance the biosynthesis of arthritic cartilage, and results in the improvement of arthritic histopathological changes. PMID- 16359877 TI - The CALM1 core promoter polymorphism is not associated with hip osteoarthritis in a United Kingdom Caucasian population. AB - OBJECTIVE: A convincing genetic association with hip osteoarthritis (OA) of a functional single nucleotide polymorphism (SNP) in the core promoter of the calmodulin 1 gene CALM1 was recently reported in a Japanese population. The T allele of the SNP encoded OA susceptibility and this was mediated by a reduced expression of CALM1. Our objective was to assess whether the SNP was also associated with hip OA in UK Caucasians. METHODS: The SNP was genotyped in 920 cases that had undergone elective joint replacement of the hip due to end-stage primary OA and in 752 age-matched controls. RESULTS: Our study had greater than 97% power to observe an effect comparable to that seen in the Japanese study. However, there was no significant difference (P< or =0.05) in genotype or allele frequencies between our cases and our controls. There was also no significant difference when the cases were stratified by sex. CONCLUSION: Our data on a cohort of 1672 individuals implies that the CALM1 core promoter polymorphism is not a risk factor for OA etiology in Caucasians. Our study does not call in to question the veracity of the Japanese report. Instead it highlights the heterogeneous nature of OA genetic susceptibility. PMID- 16359878 TI - Near-infrared spectroscopy in the routine diagnostic work-up of patients with leg ischaemia. AB - OBJECTIVE: To examine the reproducibility and clinical applicability of near infrared spectroscopy (NIRS) in patients with leg ischaemia. DESIGN: Prospective comparative diagnostic study. MATERIALS AND METHODS: Routinely measured peripheral blood pressure and microcirculatory parameters were compared with tissue oxygen saturation (TsO(2)) measurements using a new NIRS device on the calf muscle and the foot. Healthy subjects (n=20) and patients in different stages of leg ischaemia (n=45) were investigated at rest and after provocation: a treadmill test, arterial occlusion and a change in posture. Healthy volunteers were used as an age-matched reference population. RESULTS: Reproducibility of the NIRS was excellent (intraclass correlation coefficient at rest was 0.91 (95% confidence interval: 80-99). Resting TsO(2) (65%) in healthy controls did not differ significantly from that in patients (Fontaine 2: 66%; F3/4: 68%). After exercise, a significant reduction in TsO(2) was observed only in patients: Fontaine 1: 60%; Fontaine 2: 21%, Fontaine 3/4: 29%. The ankle/brachial index after exercise showed a good correlation (R=0.73) with TsO(2) at the end of the treadmill test. No correlation between NIRS and other micro- or macrocirculatory parameters was found. CONCLUSION: NIRS is a very reproducible tool to assess tissue oxygen saturation, but is not useful for the routine work-up of patients with leg ischaemia. PMID- 16359879 TI - Beta-blockers improve in-hospital and long-term survival in patients with severe left ventricular dysfunction undergoing major vascular surgery. AB - OBJECTIVES: To study whether beta-blockers reduce in-hospital and long-term mortality in patients with severe left ventricular dysfunction (LVD) undergoing major vascular surgery. DESIGN: Observational cohort study. MATERIALS: Five hundred and eleven patients with severe LVD (ejection fraction<30%) undergoing major non-cardiac vascular surgery. METHODS: In all patients, cardiac risk factors, medication (including beta-blockers), and dobutamine stress echocardiography (DSE) results were noted prior to surgery. DSE was evaluated for rest and stress-induced new wall motion abnormalities. Endpoint was in-hospital and long-term mortality. Propensity scores for beta-blockers were calculated and regression models were used to analyse the relation between beta-blockers and mortality. RESULTS: Mean age was 64+/-11 years and 383 patients (75%) were male. 139 patients (27%) used beta-blockers. Stress-induced ischemia occurred in 82 patients (16%). Median follow-up was 7 years (interquartile range: 3-10). In hospital and long-term mortality was observed in 64 (13%) and 171 (33%) patients, respectively. After adjusting for clinical variables, DSE results and propensity scores, beta-blockers were significantly associated with reduced in-hospital and long-term mortality (OR: 0.18, 95% CI: 0.04-0.74 and HR: 0.38, 95% CI: 0.22-0.65, respectively). CONCLUSION: In patients with severe LVD undergoing major vascular surgery, the use of beta-blockers is associated with a reduced incidence of in hospital and long-term postoperative mortality. PMID- 16359880 TI - Foot skin blood flow following infrainguinal revascularization for critical lower limb ischemia. AB - INTRODUCTION: The aim of this study was to assess the blood flow in the feet before and after lower limb revascularization using laser Doppler imaging (LDI). METHODS: Ten patients with critical lower limb ischemia were prospectively enrolled from June to October 2004. All patients underwent successful unilateral surgical interventions including above-knee bypass, distal bypass and endarterectomy. Skin blood flow (SBF) over the plantar surface of both forefeet and heels was measured by LDI 24h before and 10 days after revascularization, expressed in perfusion units (PU), and reported as mean+/-SD. RESULTS: Measurements in the forefoot and heel were similar. Before revascularization mean SBF was significantly lower in the ischemic foot (130+/-71 PU) compared to the contralateral foot (212+/-68 PU), p<0.05. After revascularization a significant increase of the SBF in the forefoot (from 135+/-67 to 202+/-86 PU, p=0.001) and hindfoot (from 148+/-58 to 203+/-83, p=0.001) was observed on the treatment side. However, a large decrease of the SBF was seen in forefoot and hindfoot on the untreated side (from 250+/-123 PU to 176+/-83 and from 208+/-116 to 133+/-40, p=0.001, respectively). CONCLUSION: This study confirms the benefits of revascularization in patients with nonhealing foot lesions due to critical limb ischemia. A significant increase of the SBF was observed on the treatment side. However, an unexpected decrease was observed on the untreated side. PMID- 16359881 TI - Anatomical study of the gastrocnemius venous network and proposal for a classification of the veins. AB - OBJECTIVES: To present a detailed description of the gastrocnemius venous network. DESIGN: Anatomical study in cadavers. MATERIAL AND METHODS: Forty lower limbs from 20 adult male cadavers were studied. All gastrocnemius veins were dissected from the gastrocnemius muscle heads proximally toward their drainage site. RESULTS: Eighty heads of 40 gastrocnemius muscles showed 438 gastrocnemius veins. The number of veins per muscle head varied between 2 and 12. There were 221 gastrocnemius trunks distributed as 95 main gastrocnemius trunks, 81 axial and 45 collateral ones. From the 95 main gastrocnemius trunks, 83 (87%) drained into the popliteal vein. Direct observation of the gastrocnemius venous network allowed us to classify the anatomical distribution as four distinct types. CONCLUSIONS: The majority of main gastrocnemius venous trunks drain into the popliteal vein. There is wide variability in the number of gastrocnemius veins. We propose a classification of four distinct types of anatomical pattern. PMID- 16359882 TI - Contrasting effects of varicose vein surgery on endothelial-dependent and independent cutaneous vasodilation in the perimalleolar region. AB - OBJECTIVES: To evaluate the effects of varicose vein surgery on cutaneous microvascular perfusion and vasodilator responses to acetylcholine (Ach) and sodium nitroprusside (SNP) in the gaiter area of patients with great saphenous vein insufficiency. METHODS: Twenty-nine patients with isolated great saphenous vein incompetence attended three study mornings (before surgery, and 6-8 weeks and 6 months after sapheno-femoral ligation+partial stripping) during which cutaneous microvascular responses were measured in the supine and standing positions using laser Doppler fluximetry (LDF) combined with incremental-dose iontophoretic administration of endothelial-dependent (Ach) and -independent (SNP) vasodilators. RESULTS: Varicose vein surgery had no significant effect on baseline cutaneous perfusion or the microvascular response to Ach: e.g. peak vasodilator responses to the 1000 microC stimulus were mean 58 SEM 7, 64 SEM 6 and 65 SEM 7PU on the pre-operative, 6-8 weeks and 6 months assessments. In contrast, the corresponding responses to SNP were significantly increased following surgery: e.g. at 2000 microC, mean 63 SEM 9, 142 SEM 4 and 157 SEM 9PU (p<0.0001) in the upright position. CONCLUSIONS: Sapheno-femoral ligation and partial stripping in patients with great saphenous vein insufficiency improves endothelial-independent cutaneous vasodilator function at the gaiter area, which may at least partly explain the benefits of surgery in reducing the risk of venous ulceration. PMID- 16359883 TI - Common peroneal nerve injury during varicose vein operation. AB - Common peroneal nerve (CPN) injury produces considerable and serious disability. The nerve is most frequently damaged as a result of trauma (sharp or blunt, traction, fracture, laceration, and avulsion). Less often iatrogenic injury is the cause of damage (application of tight plaster, retraction injury, division during operation). Even rarer is the complete or partial division of CPN during varicose vein operations. In the UK, on average 34 patients every year begin legal action against their medical attendants in connection with the treatment of varicose veins, on a background of an estimated 100,000 procedures performed. Nerve damage is the most frequent of all major complications that result in legal action; it is cited in 15% of cases. The commonest nerve injury, accounting for about half the cases, is to the common peroneal nerve just before or, as it crosses the neck of the fibula. We present three examples in two cases, which outline the risk of CPN injury, the spectrum of clinical presentation and the problems produced by a failure to recognise the deficit immediately. Regional anatomy, consequences of nerve damage and management options is discussed. PMID- 16359884 TI - Conversion to open repair after endografting for abdominal aortic aneurysm: causes, incidence and results. AB - OBJECTIVE: To evaluate frequency, causes and results of conversion to Open repair (OR) after endovascular repair (EVAR) in a single centre during an 8-year period. DESIGN: Six hundred and forty-nine consecutive patients undergoing EVAR were followed up prospectively for endograft-related complications. OUTCOMES: Early conversion was any OR during or within 30 days from the primary EVAR. Late conversion was any OR with removal of the endograft after 30 days since a completed EVAR procedure. RESULTS: Median patient follow-up was 38 months (1-93 months). Conversion to OR was performed in 38 patients; nine early and 29 late. Most (7/9) early conversions were due to extensive vessel calcification. Peri operative mortality was 22% (2/9). Late conversions occurred at a median of 33 months after primary EVAR: 29 were elective and 4 urgent. During the same interval, 79 secondary endovascular interventions were performed, 7 of which failed. The risk of conversion to OR was 9% at 6 years. At multivariate logistic regression analysis, no single factor (short, large or angulated neck, suprarenal fixation, large pre-operative diameter, iliac aneurysms, ASA score risk) was associated with the risk of late failure requiring conversion to OR. CONCLUSION: The risk of death after early conversion should be recognized, to avoid forcing morphological indications for primary EVAR. Occurrence of late conversion after EVAR is not negligible, affecting almost 1 out of 10 patients after 6 years. In the presence of an expanding aneurysm after EVAR, especially after a failed secondary endovascular correction, an aggressive attitude in fit patients allows outcomes at similar to those of primary OR. PMID- 16359885 TI - Remote superficial femoral endarterectomy: Long-term results. AB - PURPOSE: The aim of this study was to determine long-term results following successful remote superficial femoral endarterectomy (RSFE). METHODS: RSFE is a minimally invasive technique of revascularising the superficial femoral artery. A single incision was made over the origin of the superficial femoral artery. The endarterectomy was carried out in a closed fashion from above. The cut end of distal atheroma was secured with a stent. Following RSFE patients were followed up with intravenous digital subtraction angiography (IVDSA) and 3-monthly duplex scans. IVDSA was repeated if any abnormality was found. RESULTS: RSFE was attempted on 30 patients with 33 symptomatic legs to treat tissue loss (n = 3), rest pain (n = 3) or intermittent claudication (n = 27). In 26 limbs it was possible to complete the RSFE satisfactorily (technical success 79%), but during follow-up 18 later developed stenoses. Of 31 stenoses detected, 27 were treated by angioplasty. Primary patency at 1, 2 and 5 years was 38, 31 and 16%, respectively. Primary-assisted patency at 1, 2 and 5 years was 77, 65 and 60%. CONCLUSIONS: Primary-assisted patency following RSFE is reasonable, however, it is only achieved with life-long surveillance and intervention. Until results can be improved the widespread use of RSFE cannot be recommended. PMID- 16359886 TI - The retrojugular approach to carotid endarterectomy--a safer technique. PMID- 16359887 TI - Inflammation and atherosclerosis. AB - PURPOSE: The aim of this article is to discuss the role of inflammation in atherosclerosis. SUMMARY: An initial chemical, mechanical or immunological insult induces endothelial dysfunction. This triggers a cascade of inflammatory reactions, in which monocytes, macrophages, T lymphocytes and vascular smooth muscle cells participate. Leukocyte adhesion molecules, cytokines, growth factors and metalloproteinases participate in all stages of atherogenesis. Almost all of the traditional risk factors for atherosclerosis are associated with and participate in the inflammatory process. Many infectious agents, mainly Chlamydia pneumoniae, have been proposed as potential triggers of the cascade. The immune system has been implicated in plaque formation, through the activation of cellular and humoral immunity against innate or microbial heat shock protein 60. Methods of detection of systemic or local plaque inflammation have been developed and research is being conducted on the potential use of anti-inflammatory and antibiotic drugs in atherosclerosis. PMID- 16359888 TI - Severity of asymptomatic carotid stenosis and risk of ipsilateral hemispheric ischaemic events: results from the ACSRS study. Nicolaides et al.: EJVES 2005; 30: 275-284. PMID- 16359889 TI - Transilluminated powered phlebectomy: not enough advantages? Review of the literature. AB - BACKGROUND: Recently, new procedures for the treatment of varicose veins have been developed. The purpose of this review is to analyse the data available concerning the transilluminated powered phlebectomy (TIPP). DESIGN: Review of the English literature. RESULTS: The number of studies is limited. Currently, no trial has proven any significant advantage of TIPP technique when compared with conventional surgery, except for the number of surgical incisions. TIPP procedure seems to be shorter than conventional surgery, particularly for the extensive or recurrent varicose veins. CONCLUSIONS: Several questions regarding TIPP technique remain. Further randomised trials are needed to determine the benefit of this procedure. PMID- 16359890 TI - The combined effect of iloprost and N-acetylcysteine in preventing spinal cord ischemia in rabbits. AB - OBJECTIVES: This study investigated the cytoprotective effects of N acetylcysteine (NAC) and iloprost on spinal cord ischemia in an experimental model. MATERIALS AND METHODS: Thirty-five (male) New Zealand white rabbits were included in five study groups (n=7, each group). One group served as Sham. Rabbits in other groups had their abdominal aorta cross-clamped just above the iliac bifurcation for 40 min. During aortic cross clamping, iloprost, NAC, both iloprost and NAC or saline (control) were infused. RESULTS: In NAC, iloprost, and iloprost+NAC groups, neurological status of rabbits (Tarlov score) 24 and 48 h after the operation was better than the control group (p<0.01), but worse than the Sham group (p<0.01). There was minimal neuronal damage in the iloprost treated groups compared to the NAC group (p<0.05). Mean viability index values in NAC, iloprost and iloprost+NAC groups were higher than the control group (p<0.01). Viability index in the NAC group was lower than the iloprost and iloprost+NAC groups. CONCLUSIONS: The use of iloprost and NAC may provide better protection from spinal cord ischemia. PMID- 16359891 TI - Skeletal scintigraphy in the horse: current indications and validity as a diagnostic test. AB - Gamma scintigraphy is a well established imaging modality, which in equine practice is predominantly used in the investigation of lameness and suspected musculoskeletal problems. Whilst there is a growing literature on the indications for scintigraphy in the horse, there has been little emphasis so far on identifying the validity of scintigraphy as a diagnostic test. In particular, to what extent can the results of a scintigraphic examination be trusted? In this opinionated review the current indications for skeletal scintigraphy in the horse are considered and its validity as a diagnostic test reviewed. It is concluded that in the majority of published studies, inherent biases in study design make it difficult to assess accurately the validity of skeletal scintigraphy in the horse. Further well designed, prospective studies are required to assess the validity of skeletal scintigraphy as a diagnostic tool, particularly in horses with subtle alterations in radiopharmaceutical uptake. PMID- 16359892 TI - Immunocytochemical differentiation of neoplastic and hyperplastic canine epithelial lesions in cytologic imprint preparations. AB - The aim of the study was to test the potential use of commercially available antibodies generated against human cytokeratins in differentiating canine epithelial tumours in cytological samples. Immunocytochemical staining procedures were performed on 183 different primary epithelial canine tissues (including hyperplasia [n=7], dysplasia [n=3], benign [n=54] and malignant [n=119] neoplasia) and 20 distant metastases of 13 of the malignant tumours. All epithelial tumours and their metastases stained distinctly positive with broad spectrum anti-cytokeratin AE1/AE3. Immunocytological reactions with broad spectrum anti-cytokeratin KL1 produced less reliable results. Numerous negative reactions were found, especially in columnar epithelium tumours, whereas squamous epithelium tumours were KL1-positive. In most cases specific antibodies CK7, CK8,CK14,CK18 and CK20 showed similar reaction patterns when compared to reactivity in human tissues. Immunocytological staining was found to be a reliable and valuable diagnostic technique when combined with conventional cytology and may be especially suitable for the differentiation of undifferentiated epithelial tumours and distant metastases of unknown origin. PMID- 16359893 TI - Non-skeletal scintigraphy of the horse: indications and validity. AB - Gamma scintigraphy is an established imaging modality in the horse and is principally utilised to investigate skeletal disease using bone-seeking radiopharmaceuticals. However, depending on the radiopharmaceutical and imaging sequence, scintigraphy can be used to investigate disease in virtually any organ system. In this article the indications and validity of scintigraphy as a clinical diagnostic tool to investigate non-skeletal disease in the horse are reviewed. These indications include: investigation of dental disease, identification of inflammatory foci, vascular lesions, muscle and tendon pathology, assessment of physiological function of the lungs, gastrointestinal tract and other organs. Some of the techniques described for use in the horse have not been validated fully and as a result non-skeletal scintigraphy requires further investigation and validation using well-designed prospective studies. Such information can be used by clinicians to make informed decisions regarding the clinical and economic issues associated with a specific scintigraphic technique and may help when interpreting the results. PMID- 16359894 TI - Non-invasive assessment of airway responsiveness in healthy and allergen sensitised cats by use of barometric whole body plethysmography. AB - This study aimed at determining whether airway responsiveness (AR) tests performed by use of barometric whole body plethysmography (BWBP) were repeatable in cats and to what extent AR was affected by the nebulization protocol used, the age of the animals, the inflammatory status of the airways and prior bronchodilator treatment. Repeatability of AR was tested on two occasions in 30 healthy cats. The concentration of carbachol inducing a 300% increase of the enhanced pause (Penh)--an estimator of airflow limitation--was calculated (C Penh300) and did not differ significantly between the two tests (0.035+/-0.017% compared to 0.034+/-0.016%) and was significantly and positively correlated. The comparison between rapidly and slowly increasing carbachol concentrations was performed in ten healthy cats and showed a significantly lower C-Penh300 (%) when slowly increasing concentrations were used (0.037+/-0.013% compared to 0.039+/ 0.015%, P<0.05). A significant age-related increase of C-Penh300 was evidenced by performing AR tests in 15 healthy cats at 12, 18, 24 and 30 months (12 months: 0.026+/-0.008%, 18 months: 0.031+/-0.009%, 24 months: 0.038+/-0.01%, 30 months: 0.043+/-0.014%, P<0.05). C-Penh300 significantly decreased in 12 Ascaris suum sensitised cats after allergen exposure (0.026+/-0.016% compared to 0.033+/ 0.016%, P<0.05) and was negatively correlated with the granulocyte percentage of bronchoalveolar lavage fluid (r=-0.36, P<0.01). Compared with a placebo inhalation, pre-treatment with inhaled salbutamol significantly increased C Penh300 in four healthy cats (0.093+/-0.021% compared to 0.036+/-0.004%, P<0.05). This study provides evidence that AR determination by use of BWBP is promising as non-invasive indicator of lower airway inflammation or for monitoring response to bronchodilator treatment. PMID- 16359895 TI - Prulifloxacin: a new fluoroquinolone for the treatment of acute exacerbation of chronic bronchitis. AB - Empiric therapy with oral antibiotics is normal practice in the treatment of acute exacerbations of chronic bronchitis (AECB), but there is growing concern regarding efficacy of the currently available antimicrobials. Prulifloxacin, the lipophilic prodrug of ulifloxacin, is an oral fluoroquinolone antibacterial agent with a broad-spectrum in vitro activity against Gram-negative and -positive bacteria, and a long elimination half-life, which allows the once-daily administration. In addition, it penetrates extensively into lung tissues. Statistical analyses indicated a significant linear trend between the prulifloxacin 300, 450, and 600 mg doses, which would point to an interesting relationship between dose employed and response obtained. The 600 mg once-daily dose showed the best risk/benefit ratio, and was selected for use in the pivotal clinical trials. In well-designed clinical trials, prulifloxacin 600 mg administered once daily for 10 days in patients with AECB showed good clinical and bacteriological efficacy (similar to that of ciprofloxacin or co-amoxiclav). In particular, the clinical response rates were favourable in all clinical trials, with eradication rates in patients with pneumococcal infections at least as high as the comparators. It can be concluded that prulifloxacin 600 mg once daily is a new therapeutic prospect in the antimicrobial therapy of AECB. In particular, since good patient compliance is a key factor in the successful treatment of any infection, the once daily treatment with prulifloxacin may have some compliance advantages compared to the twice-daily treatment with agents such as ciprofloxacin or co-amoxiclav. PMID- 16359896 TI - Safety of inhaled corticosteroids: room for improvement. AB - Inhaled corticosteroids (ICS) are the standard of care in asthma and are widely used in the treatment of patients with chronic obstructive pulmonary disease. High-dose regimens and long-term use of ICS in predisposed individuals may be associated with a variety of side effects, similar to those observed with systemic corticosteroid therapy. Side effects associated with long-term ICS use include reduction in growth velocity, cataracts, glaucoma, osteoporosis, and fractures. Fear of unwanted complications may be of concern in all patients using ICS, particularly in age- and gender-specific populations that are more prone to develop side effects or to reduce treatment adherence because of physical, behavioral, or psychological problems. In addition to concerns about ICS safety, dosing regimens that are difficult to follow may further reduce a patient's ability to comply with treatment. Ciclesonide, a new-generation ICS with unique pharmacokinetic properties, was developed to provide effective anti-inflammatory control for asthma with once-daily administration to improve patient adherence and a high safety profile to reduce the occurrence of local and systemic side effects. PMID- 16359897 TI - Physiology and pathophysiology of growth hormone-binding protein: methodological and clinical aspects. AB - Circulating GH is partly bound to a high-affinity binding protein (GHBP), which in humans is derived from cleavage of the extracellular domain of the GH receptor. The precise biological function GHBP is unknown, although a regulation of GH bioactivity appears plausible. GHBP levels are determined by GH secretory status, body composition, age, and sex hormones, but the cause-effect relationships remain unclarified. In addition to the possible in vivo significance of GHBP, the interaction between GH and GHBP has methodological implications for both GH and GHBP assays. The present review concentrates on methodological aspects of GHBP measurements, GHBP levels in certain clinical conditions with a special emphasis on disturbances in the GH-IGF axis, and discusses the possible relationship between plasma GHBP and GH receptor status in peripheral tissues. PMID- 16359898 TI - Liver failure and neurologic disease in a patient with homocystinuria. AB - A homocystinuric patient with the typical clinical and metabolic features of the disorder started treatment at 22 years of age. A diet with controlled methionine intake resulted in good metabolic control for five years, but control could not be attained for the following 1 1/2 years. This was accompanied by the development of paraparesis and increasing liver failure. A liver transplantation was successful in achieving metabolic control without the need for any dietary restrictions. PMID- 16359899 TI - From an inborn error patient to a search for regulatory meaning: a biotin conducted voyage. AB - This article summarizes some findings of a research that I have pursued for the past 25 years, whose roots are immersed in the field of inherited metabolic disorders, and deal with different aspects of the vitamin biotin, starting with a patient with multiple carboxylase deficiency (MCD). Several of MCD clinical manifestations resemble those of infant malnutrition; we demonstrated that about one-third of infants with this common nutritional disorder were indeed biotin deficient, and that this deficiency is metabolically significant, by studying urine instead of blood, studying urinary organic acids by gas chromatography-mass spectrometry. Remarkably, the metabolic abnormalities became apparent only after protein feeding was started, suggesting that this phenomenon may contribute to the worsening of malnourished individuals when they are abruptly fed. Afterwards, we studied biotin deficiency at the tissue level. Carboxylase activities and masses were significantly reduced in liver, kidney, muscle, adipose tissue, intestine, and spleen, but brain and heart were spared; their mRNAs remained unchanged. On the other hand, holocarboxylase synthetase (HCS) mRNA levels were markedly low in the deficient animals, and increased upon biotin injection. Over 2000 human genes have been identified that depend on biotin for expression. To probe into the "logic" of this enigma, we have started comparative studies among evolutionarily distant organisms, such as mouse and Saccharomyces cerevisiae, and we are now looking for biotin effects on specific genes and proteins, such as HCS and hexokinases, and on their proteomes. PMID- 16359900 TI - A new diagnostic assay for glycogen storage disease type II in mixed leukocytes. AB - We have established a new method for the enzymatic diagnosis of glycogen storage disease type II (Pompe disease or acid maltase deficiency) using mixed leukocytes. The method employs glycogen and 4-methylumbelliferyl-alpha-D glucopyranoside (4MU-alphaGlc) as substrates for measuring the lysosomal acid alpha-glucosidase (acid alphaGlu) activity, and incorporates acarbose to eliminate the interference of unrelated alpha-glucosidases (predominantly maltase glucoamylase). It is shown that 3.0 micromol/L acarbose completely inhibits the maltase-glucoamylase activity at pH 4.0, but the lysosomal acid alphaGlu activity by less than 5%. With this method, we determined the acid alphaGlu activity in mixed leukocytes from 25 patients with glycogen storage disease type II (2 infantile and 23 late-onset cases), one GAA2/GAA2 homozygote and 30 healthy subjects. In the assay with glycogen as substrate, the addition of acarbose created a clear separation between the patient and the control ranges. In the assay with 4MU-alphaGlc as substrate, the two ranges were fully separated but remained very close despite the use of acarbose. The separation of the patient and normal ranges was improved considerably by taking the ratio of acarbose inhibited over uninhibited activity. A GAA2/GAA2 homozygote was correctly diagnosed with 4MU-alphaGlc but misdiagnosed as patient when glycogen was used as substrate. We conclude that the inclusion of 3.0 micromol/L acarbose in the assays with glycogen and 4MU-alphaGlc substrates at pH 4.0 allows for the specific measurement of lysosomal acid alphaGlu activity in mixed leukocytes, thus enabling a reliable diagnosis of glycogen storage disease type II in this specimen. PMID- 16359901 TI - Role of Bmi-1 and Ring1A in H2A ubiquitylation and Hox gene silencing. AB - Polycomb group (PcG) proteins exist in at least two biochemically distinct protein complexes, the EED-EZH2 complex and the PRC1 complex, that respectively possess H3-K27 methyltransferase and H2A-K119 ubiquitin E3 ligase activities. How the enzymatic activities are regulated and what their role is in Hox gene silencing are not clear. Here, we demonstrate that Bmi-1 and Ring1A, two components of the PRC1 complex, play important roles in H2A ubiquitylation and Hox gene silencing. We show that both proteins positively regulate H2A ubiquitylation. Chromatin immunoprecipitation (ChIP) assays demonstrate that Bmi 1 and other components of the two PcG complexes bind to the promoter of HoxC13. Knockout Bmi-1 results in significant loss of H2A ubiquitylation and upregulation of Hoxc13 expression, whereas EZH2-mediated H3-K27 methylation is not affected. Our results suggest that EZH2-mediated H3-K27 methylation functions upstream of PRC1 and establishes a critical role for Bmi-1 and Ring1A in H2A ubiquitylation and Hox gene silencing. PMID- 16359902 TI - Organized and highly dispersed growth of MnO2 nano-rods by sonochemical hydrolysis of Mn3acetate. AB - Highly dispersed and non-agglomerated alpha-MnO(2) nano-needles of dimensions 20 30 nm have been synthesized by the application of ultrasound radiation on the aqueous solution consisting of manganese(3)acetate close to neutral pH followed by mild drying. With a similar reaction system, hot hydrolysis (non-sonochemical process) produced beta-MnO(2) nano-rods of length 100-200 nm but with high degree agglomeration. Sonochemical cavitation phenomenon is suggested to have a pronounced effect for the formation of special phase and morphology. The effect is proved by the difference in the intermediate products which has difference in crystalinity and phase-purity. The intermediate phases are identified to be single-phase gamma-MnOOH for the non-sonochemical reaction and mixture of gamma MnOOH, alpha-MnO(2) and beta-MnO(2) for the sonochemical products. PMID- 16359903 TI - The effect of ultrasonic waves in conducting polymer solution. AB - The effects of ultrasonic wave on the conducting polymer polyaniline dissolved in DMSO were observed. The UV-visible, infrared and NMR analysis show that the polymer undergo a redox and doping transition when the ultrasound wave interacts with the polymer dissolved into the solvent. The proposed mechanism to explain these effects is based on the solvent's hygroscopicity properties. The interaction with the ultrasonic wave, homolitically dissociates the water molecule producing radical species, and these species interact with the dissolved conducting polymer changing its oxidation and doped state. The resulting effects of that interaction are the modification of oxidation state of the conducting polymer and the decrease of the amount of water molecule in the solvent. From those results, we have proposed one straightforward method to eliminate water contamination in the solvent DMSO using ultrasonic waves. PMID- 16359905 TI - Effect of surface acoustic waves on the catalytic decomposition of ethanol employing a comb transducer for ultrasonic generation. AB - The effect of surface acoustic waves, generated on a silver catalyst using a comb transducer, on the catalytic decomposition of ethanol is examined. The comb transducer employs purely mechanical means for surface acoustic wave (SAW) transduction. Unlike interdigital SAW transducers on piezoelectric substrates, the complicating effects of heat generation due to electromechanical coupling, high electric fields between adjacent electrodes, and acoustoelectric currents are avoided. The ethanol decomposition reactions are carried out at 473 K. The rates of acetaldehyde and ethylene production are retarded when acoustic waves are applied. The rates recover to varying degrees when acoustic excitation ceases. PMID- 16359904 TI - Process intensification using cavitation: optimization of oxidation conditions for synthesis of sulfone. AB - Cavitation can be effectively used for intensification of chemical reactions due to the production of free radicals and conditions of high temperatures and pressures locally. In the present work, use of cavitation for the intensification of the synthesis of sulfone has been explored. The oxidation of thioether or sulfide to synthesize corresponding sulfone with 30% H(2)O(2) as an oxidant was studied under acoustic cavitation and the results have been compared with the conventional approach based on the use of mechanical agitation. The aim has been also to optimize the different operating conditions viz. molar ratio of reactants to the oxidizing agent, type of the catalyst as well as its concentration, type of the solvent and the reactant concentration, so as to maximize the degree of intensification. It was observed that under the optimized conditions of sonication, the yield of sulfone was about five to six times higher as compared to the conventional approach of using mechanical agitation only. PMID- 16359906 TI - Investigation of an HMM/ANN hybrid structure in pattern recognition application using cepstral analysis of dysarthric (distorted) speech signals. AB - Computer speech recognition of individuals with dysarthria, such as cerebral palsy patients requires a robust technique that can handle conditions of very high variability and limited training data. In this study, application of a 10 state ergodic hidden Markov model (HMM)/artificial neural network (ANN) hybrid structure for a dysarthric speech (isolated word) recognition system, intended to act as an assistive tool, was investigated. A small size vocabulary spoken by three cerebral palsy subjects was chosen. The effect of such a structure on the recognition rate of the system was investigated by comparing it with an ergodic hidden Markov model as a control tool. This was done in order to determine if this modified technique contributed to enhanced recognition of dysarthric speech. The speech was sampled at 11 kHz. Mel frequency cepstral coefficients were extracted from them using 15 ms frames and served as training input to the hybrid model setup. The subsequent results demonstrated that the hybrid model structure was quite robust in its ability to handle the large variability and non conformity of dysarthric speech. The level of variability in input dysarthric speech patterns sometimes limits the reliability of the system. However, its application as a rehabilitation/control tool to assist dysarthric motor impaired individuals holds sufficient promise. PMID- 16359907 TI - Jumping and falling death in children, adolescents, and young adults. AB - INTRODUCTION: Evaluation of fatal jumps and falls from height in children, adolescents and young adults needs careful medico-legal investigation. The aim of this study was to examine the distribution of undetermined falling from height, accidental falling, and suicidal jumping cases between 1990-2003. METHODS: Copies of death certificates relating to falling from height and jumping deaths in Hungarian youths from the ages of 0-24 years of those resident in Hungary were obtained. RESULTS: 215 (34.1%) accidental falls, 394 (62.6%) suicidal jumping, and 21 (3.3%) undetermined intent were investigated. A significant increase was detected in suicidal jumping death in the age group of 15-19 years (OR=0.7, CI=0.49-0.98, p=0.039), and in the age group of 20-24 years (OR=0.72, CI=0.54 0.96, p=0.025) in the most recent 5 year study period. CONCLUSION: Data concerning the scene investigation, detailed forensic autopsy, and family history have important role in the differentiation of fatal jumping or falling cases. Further investigation about the characteristics injuries may help the final determination of violent death causes at youth, and more effort should be directed to an effective prevention strategy for jumping or falling death cases. PMID- 16359908 TI - The interpretation of experience and its relationship to body movement: a clinical reasoning perspective. AB - In this paper, we present findings from literature which suggests an intrinsic relationship in patients with chronic pain between the development of rigid and limited perspectives based on the interpretation of experience and the development of decreased repertoires of movement patterns. We present a research based clinical reasoning model for conceptualising the teaching of movement for patients with chronic pain and contend that therapists can intentionally teach movement using fundamentally different reasoning and learning processes. We propose that these different kinds of learning will assist clinicians to translate the findings of diverse and complex pain research to clinical practice and, in particular, the teaching of these patients both new perspectives and movement patterns. PMID- 16359909 TI - Adaptor proteins and ubiquinators in TGF-beta signaling. AB - The emergence of research analyzing the TGF-beta signaling pathway and its role in stem cell plasticity and differentiation has been a source of new insights into multiple cancers. TGF-beta signaling mediator Smads are tightly dependent on modulation by adaptor proteins, such as ELF, SARA, filamin, and crkl as well as ubiquitinators, such as PRAJA and SMURFs. Despite widespread inactivation of the TGF-beta pathway in gastrointestinal tumors, only a fraction of sporadic tumors exhibit inactivating mutations in early tumor formation, which suggests a role for the modulation of TGF-beta signals by stem/progenitor cell proteins, such as ELF and PRAJA. Delineating these key interactions of the TGF-beta signaling pathway could yield powerful new therapeutics aimed at treating hitherto difficult to treat cancers. PMID- 16359910 TI - Abiotic stress, the field environment and stress combination. AB - Farmers and breeders have long known that often it is the simultaneous occurrence of several abiotic stresses, rather than a particular stress condition, that is most lethal to crops. Surprisingly, the co-occurrence of different stresses is rarely addressed by molecular biologists that study plant acclimation. Recent studies have revealed that the response of plants to a combination of two different abiotic stresses is unique and cannot be directly extrapolated from the response of plants to each of the different stresses applied individually. Tolerance to a combination of different stress conditions, particularly those that mimic the field environment, should be the focus of future research programs aimed at developing transgenic crops and plants with enhanced tolerance to naturally occurring environmental conditions. PMID- 16359911 TI - Voluntary control of unavoidable action. AB - Goal-oriented behavior is usually portrayed to be directly related to incentive values. Underlying mechanisms are thought to include reward-oriented response bias and perceptual sensitivity. A recent electrophysiological study by Minamimoto et al. challenges this view, and reports neural activity in the thalamus that appears to counteract response bias. This type of activity might be crucial for our ability to engage voluntarily in actions that give little or no immediate return, but are necessary in the pursuit of long-term goals. PMID- 16359912 TI - In vitro testing for genotoxicity of violacein assessed by Comet and Micronucleus assays. AB - Chromobacterium violaceum is a Gram (-) bacteria found in water samples and soils from tropical and subtropical regions of the world. Violacein, the major pigment produced by these bacteria, has been shown to have antibiotic, antitumoral and trypanocidal activities. In the present work, the genotoxicity of violacein was investigated in four different cell lines by using the alkaline Comet assay and in VERO cells using the Micronucleus test. In the alkaline Comet assay, violacein, when tested at concentrations ranging from 0.19 to 1.5 microM, did not induce a significant increase in DNA damage in HEp-2 and MA104 cells. However, violacein was positive for DNA damage in FRhK-4 cells and for both DNA damage and micronuclei in VERO cells, in a concentration-response relationship. The results of this study indicated that violacein is genotoxic in VERO and FRhK-4 cells. These findings contribute to a comprehensive evaluation of the pharmacological potential of violacein. PMID- 16359913 TI - iMARS--mutation analysis reporting software: an analysis of spontaneous cII mutation spectra. AB - The sensitivity of any mutational assay is determined by the level at which spontaneous mutations occur in the corresponding untreated controls. Establishing the type and frequency at which mutations occur naturally within a test system is essential if one is to draw scientifically sound conclusions regarding chemically induced mutations. Currently, mutation-spectra analysis is laborious and time consuming. Thus, we have developed iMARS, a comprehensive mutation-spectrum analysis package that utilises routinely used methodologies and visualisation tools. To demonstrate the use and capabilities of iMARS, we have analysed the distribution, types and sequence context of spontaneous base substitutions derived from the cII gene mutation assay in transgenic animals. Analysis of spontaneous mutation spectra revealed variation both within and between the transgenic rodent test systems Big Blue Mouse, MutaMouse and Big Blue Rat. The most common spontaneous base substitutions were G:C-->A:T transitions and G:C- >T:A transversions. All Big Blue Mouse spectra were significantly different from each other by distribution and nearly all by mutation type, whereas the converse was true for the other test systems. Twenty-eight mutation hotspots were observed across all spectra generally occurring in CG, GA/TC, GG and GC dinucleotides. A mutation hotspot at nucleotide 212 occurred at a higher frequency in MutaMouse and Big Blue Rat. In addition, CG dinucleotides were the most mutable in all spectra except two Big Blue Mouse spectra. Thus, spontaneous base-substitution spectra showed more variation in distribution, type and sequence context in Big Blue Mouse relative to spectra derived from MutaMouse and Big Blue Rat. The results of our analysis provide a baseline reference for mutation studies utilising the cII gene in transgenic rodent models. The potential differences in spontaneous base-substitution spectra should be considered when making comparisons between these test systems. The ease at which iMARS has allowed us to carry out an exhaustive investigation to assess mutation distribution, mutation type, strand bias, target sequences and motifs, as well as predict mutation hotspots provides us with a valuable tool in helping to distinguish true chemically induced hotspots from background mutations and gives a true reflection of mutation frequency. PMID- 16359914 TI - Conformational preferences and orbital interactions for methyl haloacetates. AB - Conformational preferences and orbital interactions of methyl chloroacetate (1), methyl bromoacetate (2) and methyl iodo-acetate (3) were analyzed using experimental infra-red data, theoretical calculations and NBO analyses. The conformational equilibria of compounds 1-3 can be represented by their cis and gauche rotamers. The gauche form of 1 is stable in the vapour phase and in a non polar solvent, but the cis is predominant in a polar solvent. For 2 the gauche form is more stable than the cis, in both the vapour and liquid phases, but for compound 3 only the gauche form was observed both in vapour phase as in solution. These conformational preferences were attributed to the orbital interaction between two antibonding orbitals pi(C=O)(*)-->sigma(C-X)(*). This unexpected interaction was possibly due to the high (0.2) electron density on pi(C=O)(*), which results from the interaction between ether oxygen lone pair and pi(C=O)(*). PMID- 16359915 TI - Catalytic kinetic spectrophotometry for the determination of trace amount of oxalic acid in biological samples with oxalic acid-rhodamine B-potassium dichromate system. AB - A new catalytic kinetic spectrophotometric method was proposed for determining trace oxalic acid based on the catalytic effect of oxalate on the oxidation of potassium dichromate with rhodamin B in 0.10 M of sulfuric acid. Good linearity is obtained over the concentration range 0.40-6.0 microg/mL of oxalic acid. After the reactions of the catalytic and non-catalytic systems were terminated by using 2.00 mL of 4 M sodium hydroxide solution, they can be stable for 3 h at room temperature. The apparent activation energy of the catalytic reaction is 12.44 kJ/mol. The effect of 50 coexisting substances was observed. The method was used to determine trace oxalic acid in tea, spinach and urine samples with satisfactory results. PMID- 16359916 TI - Development of SOVAT: a numerical-spatial decision support system for community health assessment research. AB - INTRODUCTION: The development of numerical-spatial routines is frequently required to solve complex community health problems. Community health assessment (CHA) professionals who use information technology need a complete system that is capable of supporting the development of numerical-spatial routines. BACKGROUND: Currently, there is no decision support system (DSS) that is effectively able to accomplish this task as the majority of public health geospatial information systems (GIS) are based on traditional (relational) database architecture. On Line Analytical Processing (OLAP) is a multidimensional data warehouse technique that is commonly used as a decision support system in standard industry. OLAP alone is not sufficient for solving numerical-spatial problems that frequently occur in CHA research. Coupling it with GIS technology offers the potential for a very powerful and useful system. METHODOLOGY: A community health OLAP cube was created by integrating health and population data from various sources. OLAP and GIS technologies were then combined to develop the Spatial OLAP Visualization and Analysis Tool (SOVAT). RESULTS: The synergy of numerical and spatial environments within SOVAT is shown through an elaborate and easy-to-use drag and drop and direct manipulation graphical user interface (GUI). Community health problem solving examples (routines) using SOVAT are shown through a series of screen shots. DISCUSSION: The impact of the difference between SOVAT and existing GIS public health applications can be seen by considering the numerical-spatial problem-solving examples. These examples are facilitated using OLAP-GIS functions. These functions can be mimicked in existing GIS public applications, but their performance and system response would be significantly worse since GIS is based on traditional (relational) backend. CONCLUSION: OLAP-GIS system offer great potential for powerful numerical-spatial decision support in community health analysis. The functionality of an OLAP-GIS system has been shown through a series of example community health numerical-spatial problems. Efforts are now focused on determining its usability during human-computer interaction (HCI). Later work will focus on performing summative evaluations comparing SOVAT to existing decision support tools used during community health assessment research. PMID- 16359917 TI - Timing of interferon therapy and sources of infection in patients with acute hepatitis C. AB - BACKGROUND/AIMS: Controversy over the selection of patients and optimum therapeutic method for acute hepatitis C has continued. The aims of this study were to investigate the source of infection, and to evaluate the timing of interferon (IFN) therapy in patients with acute hepatitis C in Japan. METHODS: The records of 102 patients from 12 facilities in Japan who developed acute hepatitis C after 1990 were investigated. In the patients treated with IFN, we performed multivariate analysis to investigate factors related to sustained virological response (SVR). RESULTS: Medical procedure was the most common source of infection, accounting for 32.4% in the 102 patients (33/102). Of 81 patients treated with IFN, 71 patients were followed after IFN therapy, and 57/71 (80.3%) had SVR. The SVR rate was significantly higher in patients treated with IFN within 24 weeks from onset of symptoms than the SVR rate in those treated after 25 weeks (P=0.0016). Multivariate analysis revealed that only the duration between onset of symptoms and initiation of IFN therapy (within 24 weeks) was related to SVR. CONCLUSIONS: Our multicenter cooperative survey revealed that medical procedure was the most frequent source of infection in acute hepatitis C. As concerns the therapy, interferon treatment should be initiated within 24 weeks after onset of symptoms. PMID- 16359918 TI - Glutamate-based therapeutic approaches: clinical trials with NMDA antagonists. AB - The majority of clinical trials of N-methyl d-aspartate antagonists have been conducted in the fields of stroke, traumatic brain injury (TBI) and dementia. Stroke and TBI trials have involved more than 9000 patients, but have yielded no therapeutically useful agents, with the possible exception of magnesium for treatment of subarachnoid haemorrhage. Several of the synthetic N-methyl D aspartate antagonist development programmes have been abandoned owing to concerns about drug toxicity, particularly in stroke. Systematic reviews in stroke and TBI have shown that definitive conclusions cannot be drawn for most agents owing to early termination of trials. In dementia, memantine has shown some benefit in moderate-to-severe Alzheimer's disease, with no clear benefit to date for milder stages of Alzheimer's disease or for vascular dementia. Other therapeutic areas of promise remain inadequately explored at present. PMID- 16359919 TI - GABA-A receptors: a viable target for novel anxiolytics? AB - Benzodiazepine (BZ) anxiolytics mediate their clinical effects by enhancing the effect of gamma-aminobutyric acid (GABA) at the GABA-A receptor. Classical BZ full agonists such as diazepam, which maximally enhance the function of GABA-A receptors, are effective anxiolytics but carry unwanted side effects including sedation, dependence and abuse liability, limiting their utility. Although a second generation of 'partial agonist' BZs have been pursued, promising preclinical data, in terms of anxiolytic efficacy and decreased unwanted effects, have so far failed to translate to the clinic. Following the insights into GABA-A receptor subtypes mediating the effects of BZs, a third generation of 'receptor subtype-selective' BZ site ligands have been developed. However, it remains to be determined whether promising preclinical data are recapitulated in the clinic. PMID- 16359920 TI - Functional implications of neurotransmitter co-release: glutamate and GABA share the load. AB - For decades it has been thought that a neuron releases only one classical neurotransmitter from all of its processes. However, recent work has shown that most neuronal populations release more than one classical transmitter, and indeed that the transmitters can be segregated into different processes of the same neuron. Glutamate and gamma-aminobutyric acid, the major excitatory and inhibitory neurotransmitters in the mammalian central nervous system, appear to be co-released with most other transmitters, as well as with each other. The release of multiple transmitters by the same neuron enhances the spatial and temporal control of synaptic transmission. Moreover, dynamic regulation of neurotransmitter phenotypes increases the plasticity of neurotransmission, indicating potential avenues for therapeutic intervention. PMID- 16359921 TI - A derivative of Mycobacterium smegmatis mc(2)155 that lacks the duplicated chromosomal region. AB - The genome of Mycobacterium smegmatis mc(2)155 contains a 56kb duplicated region. We isolated a mutant of mc(2)155 lacking this duplication (DeltaDRKIN). This mutation did not affect the growth rate, surface properties or transformation efficiency of the organism, confirming the potential utility of DeltaDRKIN for the study of genes contained within the duplicated region. PMID- 16359922 TI - Animal models of cavitation in pulmonary tuberculosis. AB - Transmission of tuberculosis occurs with the highest frequency from patients with extensive, cavitary, pulmonary disease and positive sputum smear microscopy. In animal models of tuberculosis, the development of caseous necrosis is an important prerequisite for the formation of cavities although the immunological triggers for liquefaction are unknown. We review the relative merits and the information gleaned from the available animal models of pulmonary cavitation. Understanding the host-pathogen interaction important to the formation of cavities may lead to new strategies to prevent cavitation and thereby, block transmission. PMID- 16359923 TI - Assessment of cognitive function in heart failure patients. AB - BACKGROUND: Research on the cognitive capacity of heart failure patients is limited, with a paucity of benchmark information available for this population. It is highly likely that cognitive deficits affect patients' understanding of disease and treatment requirements, as well as limiting their functional capacity and ability to implement treatment plans, and undertake self-care. AIMS: The purpose of this study was to establish a comprehensive neurocognitive profile of the heart failure patient through systematic neurocognitive assessment and to determine whether an association existed between severity of heart failure and cognitive abilities. METHODS: Thirty-eight patients were recruited from the heart failure patient databases of two metropolitan hospitals in Melbourne, Australia. Participants were individually assessed using four standardised, internationally recognised neuropsychological tests that examined current and premorbid intelligence, memory and executive functioning. RESULTS: Although there was no significant decline from premorbid general intellectual function, other specific areas of deficit, including impaired memory and executive functioning, were identified. There were no significant correlations between heart failure severity and the neurocognitive measures used. CONCLUSION: The results support the need to recognise cognitive impairment in people with heart failure and to develop an abbreviated method of assessing cognitive function that can be easily implemented in the clinical setting. Identifying cognitive deficits in this population will be useful in guiding the content and nature of treatment plans to maximise adherence and minimise worsening of heart failure symptoms. PMID- 16359924 TI - Tools of the trade. AB - An unhurried, deft, economical technique allied to efficient use of available resources, surgical instruments and support staff is the hallmark of a 'good' surgeon. To achieve this sense of fluency, surgeons need to be familiar with the tools of their profession so that these can be employed appropriately for the task at hand. The range of instruments available in obstetrics and gynaecology is no less wide or complex than in any other branch of surgery, but formal teaching in the full range of surgical hardware is frequently lacking and an individual's exposure is generally limited to the instruments used in his or her own training hospital. This chapter sets out to describe key technical areas with which surgeons need to be familiar. PMID- 16359925 TI - Worldwide burden of gynaecological cancer: the size of the problem. AB - The estimation of cancer burden is valuable to set up priorities for disease control. The comprehensive global cancer statistics from the International Agency for Research on Cancer indicate that gynaecological cancers accounted for 19% of the 5.1 million estimated new cancer cases, 2.9 million cancer deaths and 13 million 5-year prevalent cancer cases among women in the world in 2002. Cervical cancer accounted for 493 000 new cases and 273 000 deaths; uterine body cancer for 199 000 new cases and 50 000 deaths; ovarian cancer for 204 000 new cases and 125 000 deaths; cancers of the vagina, vulva and choriocarcinoma together constituted 45 900 cases. More than 80% of the cervical cancer cases occurred in developing countries and two-thirds of corpus uteri cases occurred in the developed world. Political will and advocacy to invest in healthcare infrastructure and human resources to improve service delivery and accessibility are vital to reduce the current burden in low- and medium-resource countries. PMID- 16359926 TI - HPV testing in cervical screening. AB - High-risk human papillomavirus (hrHPV) bearing cervical intraepithelial neoplasia (CIN) is considered, as the real precursor lesion of cervical cancer and persistence of an hrHPV infection is necessary for the progression to cervical cancer. This knowledge warrants the use of hrHPV testing as an adjunct to cervical cytology in population-based screening programmes and for monitoring therapy efficacy of high-grade CIN lesions. Replacement of cytology by hrHPV testing altogether is considered, but for this to be (cost-) effective, accurate information about the specificity of the hrHPV test is required. Additional test systems that can be used to stratify women with a positive hrHPV test are HPV genotyping, viral load analysis and hrHPV mRNA analysis. The need for HPV genotyping of cervical smears is illustrated by the increased risk for high-grade cervical lesions associated with HPV types 16 and 18. In particular, for women who have normal but persistently (>1 year) HPV18-positive smears, endocervical curettage is suggested (evidently considering the age and possible future pregnancies of the respective woman) because HPV18 is associated with glandular lesions in the cervix, which are difficult to detect by cytology. PMID- 16359927 TI - The decay of memory between delayed and long-term recall in patients with temporal lobe epilepsy. AB - OBJECTIVES: Impairment of long-term recall may worsen everyday functioning of patients with epilepsy even if the standard short-term or delayed recall tests do not show significant abnormalities. We evaluated prospectively the decay of memory between delayed and long-term recall in patients with temporal lobe epilepsy (TLE) and controls with the aim of identifying the determinants of long term memory impairment. METHODS: Seventy patients with TLE and 59 controls underwent neuropsychological assessment of verbal and nonverbal memory, attention, and executive functions at visit 1. Long-term verbal and nonverbal memory was tested with the same word list, verbal logical story, and Rey Osterrieth complex figure test 4 weeks later at visit 2. The decay in memory was estimated as information recalled at visit 2 as a percentage of the delayed recall at visit 1. RESULTS: Frequent seizures (> or = 4 per month) during the study period were related to poor long-term recall, even for those patients who did relatively well on delayed recall tests. On all long-term memory tests, patients with complex partial and/or secondary generalized seizures did significantly worse than patients with simple partial seizures. The presence of interictal generalized or focal temporal epileptiform activity was associated with more accelerated forgetting of the word list and complex figure. Multiple regression analysis confirmed that number of complex partial seizures, age of patient, and abnormal interictal EEG are significant predictors of accelerated forgetting. CONCLUSIONS: Uncontrolled seizures, especially with ictal impairment of consciousness, can be a significant factor in the accelerated decay of memory, although subclinical interictal epileptiform EEG activity may also be relevant. PMID- 16359928 TI - Natural language processing to extract medical problems from electronic clinical documents: performance evaluation. AB - In this study, we evaluate the performance of a Natural Language Processing (NLP) application designed to extract medical problems from narrative text clinical documents. The documents come from a patient's electronic medical record and medical problems are proposed for inclusion in the patient's electronic problem list. This application has been developed to help maintain the problem list and make it more accurate, complete, and up-to-date. The NLP part of this system analyzed in this study-uses the UMLS MetaMap Transfer (MMTx) application and a negation detection algorithm called NegEx to extract 80 different medical problems selected for their frequency of use in our institution. When using MMTx with its default data set, we measured a recall of 0.74 and a precision of 0.756. A custom data subset for MMTx was created, making it faster and significantly improving the recall to 0.896 with a non-significant reduction in precision. PMID- 16359929 TI - Representation in stochastic search for phylogenetic tree reconstruction. AB - Phylogenetic tree reconstruction is a process in which the ancestral relationships among a group of organisms are inferred from their DNA sequences. For all but trivial sized data sets, finding the optimal tree is computationally intractable. Many heuristic algorithms exist, but the branch-swapping algorithm used in the software package PAUP* is the most popular. This method performs a stochastic search over the space of trees, using a branch-swapping operation to construct neighboring trees in the search space. This study introduces a new stochastic search algorithm that operates over an alternative representation of trees, namely as permutations of taxa giving the order in which they are processed during stepwise addition. Experiments on several data sets suggest that this algorithm for generating an initial tree, when followed by branch-swapping, can produce better trees for a given total amount of time. PMID- 16359930 TI - Pharmacogenetics: progress, pitfalls and clinical potential for coronary heart disease. AB - Much has been written about the potential of pharmacogenetic testing to inform therapy based on an individual's genetic makeup, and to decide the most effective choice of available drugs, or to avoid dangerous side effects. Currently, there is little hard data for either in the field of cardiovascular disease. The usual approach has been opportunistic use of drug trials in unrelated patients, and to look for differences in response or outcome by "candidate gene" genotype, for example genes coding for drug metabolising enzymes (activators and metabolisers), and enzymes and receptors involved in lipid metabolism, adrenergic response, etc. As with all association studies, initially promising results have often failed the test of replication in larger studies, and the relationship between the CETP Taq-I variant and response to statins has now been disproved. The strongest data to date is the report [Chasman, D.I., Posada, D., Subrahmanyan, L., Cook, N.R., Stanton Jr., V.P., Ridker, P.M., 2004. Pharmacogenetic study of statin therapy and cholesterol reduction. J. Am. Med. Assoc. 291, 2821-2827] of a poorer cholesterol-lowering response to Pravastatin in the 7% of patients carrying a certain haplotype of the HMG CoA reductase gene (14% fall versus 19%), but if this is overcome simply by a higher dose, it is of little clinical relevance. Currently, the best example of avoiding side effects is determining genotype at the CYP2C9 locus with respect of warfarin treatment, since carriers for functional variants (>20% of the population) require lower doses for optimal anticoagulation, and homozygotes, although rare, may well experience serious bleeding if given a usual dose. The full potential of this field will only be realised with much further work. PMID- 16359931 TI - Generation of mutator mutants during carcinogenesis. AB - Mutations are rare in normal cells. In contrast, multiple mutations are characteristic in most tumors. Previously we proposed a "mutator phenotype" hypothesis to explain how pre-cancer cells may acquire large number of mutations during carcinogenesis. Here we extend the "mutator phenotype" hypothesis considering recently discovered biochemical activities whose aberrant expression may result in genome-wide random mutations. The scope of this article is to emphasize that simple random point mutations can drive carcinogenesis and highlight new emerging pathways that generate these mutations. We focus specifically on random point mutations generated by replication errors, oxidative base damage, covalent base modifications by enzymes, and spontaneously generated abasic sites as a source of mutator mutants. PMID- 16359932 TI - Determination of the novel non-peptidic HIV-protease inhibitor tipranavir by HPLC UV after solid-phase extraction. AB - An HPLC method previously described for the assay of amprenavir (APV), ritonavir (RTV), indinavir (IDV), saquinavir (SQV), nelfinavir (NFV), lopinavir (LPV), atazanavir (ATV), nevirapine (NVP) and efavirenz (EFV) can be also conveniently applied, with minor gradient program adjustment, for the determination of the novel non-peptidic HIV protease inhibitor tipranavir (TPV) in human plasma, by off-line solid-phase extraction (SPE) followed by HPLC coupled with UV-diode array detection (DAD). After viral inactivation by heat, the plasma is diluted with phosphate buffer (pH 7), and subjected to a SPE on a C18 cartridge. Matrix components are eliminated with a solution of 0.1% H3PO4 solution neutralised to pH 7, and TPV is eluted with MeOH. The resulting eluate is evaporated and reconstituted in 100 microl MeOH/H2O 50/50. A 40 microl volume is injected onto a Nucleosil C18 AB column and TPV is analysed by UV detection at 201 nm using a gradient elution program constituted of MeCN and phosphate buffer adjusted to pH 5.12 and containing 0.02% sodium heptanesulfonate. The calibration curves are linear up to 75 microg/ml, with a lower limit of quantification of 0.125 microg/ml. The mean absolute recovery of TPV is 77.1+/-4.0%. The method is precise with mean inter-day coefficient of variations (CVs) within 2.2-3.4%, and accurate (range of inter-day deviations from 0.7 to 1.2%). The method has been validated and is currently applied to the monitoring of TPV plasma levels in HIV patients. PMID- 16359934 TI - Quantitation of salbutamol in human urine by liquid chromatography-electrospray ionization mass spectrometry. AB - A sensitive and specific liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) is described for quantitation of salbutamol in human urine using nadolol as the internal standard (I.S.). Urine samples were hydrolyzed with beta-glucuronidase followed by a solid-phase extraction procedure using Bond Elut-Certify cartridges. The HPLC column was an Agilent Zorbax SB C(18) column. A mixture of 0.01 M ammonium formate buffer (pH 3.5)-acetonitrile (85:15, v/v) was used as the mobile phase. Analytes were quantitated using positive electrospray ionization in a quadrupole spectrometer. Selected ion monitoring (SIM) mode was used to monitor m/z 166 for salbutamol and m/z 310 for I.S. Good linearity was obtained in the range of 10.0-2000.0 ng/ml. The limit of quantification was 10.0 ng/ml. The intra- and inter-run precision, calculated from quality control (QC) samples was less than 7.3%. The accuracy as determined from QC samples was within +/-2.6%. The method was applied for determining excretion curves of salbutamol. PMID- 16359933 TI - Direct analysis of bromide in human serum by capillary electrophoresis. AB - The purpose of the present work was the development and validation of a simple, rapid and reliable method for direct bromide quantification in serum based on capillary electrophoresis (CE). The analysis was carried out with an automated capillary electropherograph. Analytical conditions were as follows. Capillary: uncoated fused silica, effective length 50 cm, internal diameter 50 microm; voltage: 20 kV in reverse polarity mode; temperature: 25 degrees C; running buffer: 90 mmol/L sodium tetraborate decahydrate and 10 mmol/L NaCl, pH 9.24; detection: direct UV absorption at 200 nm; sample treatment: dilution of serum 1:10 with the internal standard solution (2 mmol/L thiocyanate). Under the described conditions, bromide ions and internal standard were baseline separated in 7 min. No interferences from other serum components were observed. The analytical sensitivity was characterized by a LOD: 0.05 mmol/L and a LOQ of 0.1 mmol/L. Excellent linearity was verified in the range from 2.5 to 60 mmol/L [y = 0.0746x - 0.0372; R2 = 0.9995 (x = bromide concentration; y = bromide peak area/internal standard (I.S.) peak area)]. Quantitative imprecision in intra-day (n = 7) and day-to-day (n = 7) experiments was always within R.S.D. values <2%. Recovery was quantitative throughout the range of linearity of the method. Clinical cases of infants undergoing potassium bromide therapy for refractory epilepsy were analyzed with results in agreement with literature data. On the basis of these considerations, capillary electrophoresis can be proposed as the method of choice for bromide analysis in serum samples, especially for therapeutic drug monitoring purposes. PMID- 16359935 TI - Geographical height variation among Ohio Caucasian male convicts born 1780-1849. AB - To more completely answer questions regarding health in the past, social scientists are utilizing previously underutilized data sources. This study focuses on one such source: penitentiary records, in order to examine geographical variation in height from a sample of 2554 male Caucasian convicts aged 23-45 years. Data collected include height, age, ancestry, nationality, and year and place of birth. Birth places were divided into five geographical areas corresponding roughly to the United States census geography divisions. Average stature for this sample was 68.4+/-2.5 in. (173.7+/-6.4 cm). Regression analyses on height, birth cohort, ancestry, nationality, and place of birth indicate significant but small differences in height across geographical areas. No significant differences were found due to birth cohort. PMID- 16359936 TI - Physical stature of adult Tsimane' Amerindians, Bolivian Amazon in the 20th century. AB - We examine the association between exposure to the market and Western society on the height of adult Tsimane', a foraging-farming society in the Bolivian Amazon. As with other contemporary native peoples, we find little evidence of a significant secular change in height during 1920-1980. Female height bore a positive association with own schooling and fluency in spoken Spanish and with maternal modern human capital (schooling, writing ability, and fluency in spoken Spanish), but male heights bore no association with parental height or with modern human capital. The absence of a secular change likely reflects the persistence of traditional forms of social organization and production that protect health. PMID- 16359937 TI - Instability prolongs the chondral phase during bone healing in sheep. AB - In this sheep study, we investigated the influence of fixation stability on the temporal and spatial distribution of tissues in the fracture callus. As the initial mechanical conditions have been cited as being especially important for the healing outcome, it was hypothesized that differences in the path of healing would be seen as early as the initial phase of healing. Sixty-four sheep underwent a mid-shaft tibial osteotomy that was treated with either a rigid or a semi-rigid external fixator. Animals were sacrificed at 2, 3, 6 and 9 weeks postoperatively and the fracture calluses were analyzed using radiological, biomechanical and histological techniques. Statistical comparison between the groups was performed using the Mann-Whitney U test for unpaired non-parametric data. In the callus of the tibia treated with semi-rigid fixation, remnants of the fracture haematoma remained present for longer, although new periosteal bone formation during early healing was similar in both groups. The mechanical competence of the healing callus at 6 weeks was inferior compared to tibiae treated with rigid fixation. Semi-rigid fixation resulted in a larger cartilage component of the callus, which persisted longer. Remodeling processes were initiated earlier in the rigid group, while new bone formation continued throughout the entire investigated period in the semi-rigid group. In this study, evidence is provided that less rigid fixation increased the time required for healing. The process of intramembranous ossification appeared during the initial stages of healing to be independent of mechanical stability. However, the delay in healing was related to a prolonged chondral phase. PMID- 16359938 TI - The connexins, Part III. PMID- 16359939 TI - Connexins, innexins and pannexins: bridging the communication gap. PMID- 16359940 TI - Regulation of connexin expression. AB - Gap junctions contain cell-cell communicating channels that consist of multimeric proteins called connexins and mediate the exchange of low-molecular-weight metabolites and ions between contacting cells. Gap junctional communication has long been hypothesized to play a crucial role in the maintenance of homeostasis, morphogenesis, cell differentiation, and growth control in multicellular organisms. The recent discovery that human genetic disorders are associated with mutations in connexin genes and experimental data on connexin knockout mice have provided direct evidence that gap junctional communication is essential for tissue functions and organ development. Thus far, 21 human genes and 20 mouse genes for connexins have been identified. Each connexin shows tissue- or cell type-specific expression, and most organs and many cell types express more than one connexin. Cell coupling via gap junctions is dependent on the specific pattern of connexin gene expression. This pattern of gene expression is altered during development and in several pathological conditions resulting in changes of cell coupling. Connexin expression can be regulated at many of the steps in the pathway from DNA to RNA to protein. However, transcriptional control is one of the most important points. In this review, we summarize recent knowledge on transcriptional regulation of connexin genes by describing the structure of connexin genes and transcriptional factors that regulate connexin expression. PMID- 16359941 TI - Gap junctions in skeletal development and function. AB - Gap junctions play a critical role in the coordinated function and activity of nearly all of the skeletal cells. This is not surprising, given the elaborate orchestration of skeletal patterning, bone modeling and subsequent remodeling, as well as the mechanical stresses, strains and adaptive responses that the skeleton must accommodate. Much remains to be learned regarding the role of gap junctions and hemichannels in these processes. A common theme is that without connexins none of the cells of bone function properly. Thus, connexins play an important role in skeletal form and function. PMID- 16359942 TI - Involvement of gap junctional communication in secretion. AB - Glands were the first type of tissues in which the permissive role of gap junctions in the cell-to-cell transfer of membrane-impermeant molecules was shown. During the 40 years that have followed this seminal finding, gap junctions have been documented in all types of multicellular secretory systems, whether of the exocrine, endocrine or pheromonal nature. Also, compelling evidence now indicates that gap junction-mediated coupling, and/or the connexin proteins per se, play significant regulatory roles in various aspects of gland functions, ranging from the biosynthesis, storage and release of a variety of secretory products, to the control of the growth and differentiation of secretory cells, and to the regulation of gland morphogenesis. This review summarizes this evidence in the light of recent reports. PMID- 16359943 TI - Defective gap junctional intercellular communication in the carcinogenic process. AB - Gap junctions are membrane structures made of intercellular channels which permit the diffusion from cytoplasm to cytoplasm of small hydrophilic molecules. Nearly 40 years ago, the loss of functional gap junctions has been described in cancer cells and led to the hypothesis that such type of intercellular communication is involved in the carcinogenesis process. From this time, a lot of data has been accumulated confirming that gap junctions are frequently decreased or absent in cancer cells whatever their tissue and species origins. Here, we review such data by insisting on the possible links existing between altered gap-junctional intercellular communication capacity (or the altered expression of their constitutive proteins, the connexins) and the stages of cancer progression in various cancer models. Then, we analyse particular aspects of the disturbance of connexin-mediated communication in cancer such as the cytoplasmic localization of connexins, the lack of heterologous communication between cancer cells and normal cells, the role of connexin gene mutations in cancer. In a separate part of the review, we also analyse the disturbance of gap-junctional intercellular communication during the late stages of cancer (invasion and metastasis processes). PMID- 16359944 TI - Innovations in fertility preservation for patients with gynecologic cancers. AB - OBJECTIVE: To review options for fertility preservation in women with gynecologic cancers. DESIGN: Literature review. RESULT(S): We discuss the data regarding cancer treatment and fertility outcomes and current controversies for women with gynecologic cancers. CONCLUSION(S): Gynecologic cancers represent 12%-15% of cancers affecting women, and 21% of these are diagnosed in women of reproductive age. Current advances in our understanding of these diseases, along with improved multimodality treatment, allow for consideration of fertility options. For some women with gynecologic cancers, fertility-sparing treatment might be appropriate. PMID- 16359945 TI - Role of laparoscopic treatment of endometriosis in patients with failed in vitro fertilization cycles. AB - OBJECTIVE: To report our experience in patients with previous IVF failures who conceived after laparoscopic treatment of endometriosis. DESIGN: Retrospective case series. SETTING: Tertiary center IVF and endoscopy programs. PATIENT(S): Infertility patients with history of prior IVF failures. INTERVENTION(S): Laparoscopic evaluation and treatment of endometriosis by the same surgeon. MAIN OUTCOME MEASURE(S): Occurrence of conception after laparoscopic treatment of endometriosis. RESULT(S): Of 29 patients with prior IVF failures, 22 conceived after laparoscopic treatment of endometriosis, including 15 non-IVF pregnancies and 7 IVF pregnancies. CONCLUSION(S): In the absence of tubal occlusion or severe male factor infertility, laparoscopy may still be considered for the treatment of endometriosis even after multiple IVF failures. PMID- 16359946 TI - Subfertility, fecundability, and the impact of laparoscopy on conception rates. AB - Confusion is common when trying to compare the outcome of different fertility treatments. Calculation of fecundability is an excellent adjunct to reports that show cumulative conception totals after a period of observation or treatment. PMID- 16359947 TI - Challenges of evaluating surgical outcomes. AB - The role of surgery vs. IVF-embryo transfer to improve the potential for conception will vary as enhancements of these interventions occurs. Well-designed and conducted studies, controlling for the complexities of surgical trials are needed. PMID- 16359948 TI - Laparoscopy, in vitro fertilization, and endometriosis: an enigma. AB - Studies on the respective roles of laparoscopic surgery and IVF in infertile patients with endometriosis and endometriomas are difficult to perform. Appropriate patient management is complex and requires a sophisticated understanding of individualized evidence-based decision making, surgical judgment and technical skills for laparoscopy, and thoughtful utilization of IVF technologies. PMID- 16359949 TI - Is laparoscopy back? AB - Should surgical treatment of presumed endometriosis be advocated for all patients who fail to conceive after multiple IVF cycles? Without appropriately designed clinical studies, there is currently little evidence to support this approach. PMID- 16359950 TI - The dilemma of endometriosis: is consensus possible with an enigma? AB - Many will agree that the use of laparoscopy to diagnose and potientially treat endometriosis in patients who suffer from infertility has been superseded by IVF and sometimes oocyte donation, especially in older patients. The findings of our study add another dimension to management of endometriosis in the setting of infertility and emphasize the importance of keeping laparoscopy in the infertility management equation. PMID- 16359951 TI - A critique of the Women's Health Initiative hormone therapy study. AB - OBJECTIVE: This review critiques The Women's Health Initiative (WHI) study, focusing on aspects of the study design contributing to the adverse events resulting in the study's discontinuation. CONCLUSION(S): Two aspects of the design contributed to the adverse events: [1] The decision to administer continuous combined conjugated equine estrogen (CEE)/medroxyprogesterone acetate (MPA) or E alone as a standard regimen to a population with little previous hormonal treatment, ranging in age from 50-79 years, who, because of their age, were predisposed to coronary and cerebral atherosclerosis. [2] Selection of an untested regimen of continuous combined CEE plus MPA, which we hypothesize, negated the protective effect of E on the cardiovascular and cerebrovascular systems. Multiple observational studies that preceded the WHI study concluded that the use of E alone and E plus cyclic (not daily) progestin combination treatments initiated in early menopause had beneficial effects. The therapeutic regimens resulted in prevention of atherosclerosis and reductions in coronary artery disease mortality. It is our conclusion that the WHI hormonal replacement study had major design flaws that led to adverse conclusions about the positive effects of hormone therapy. An alternative hormonal regimen is proposed that, on the basis of data supporting its beneficial cardiovascular effects, when initiated appropriately in a population of younger, more recently menopausal women, has promise to yield a more favorable risk/benefit outcome. PMID- 16359952 TI - Spontaneous embryonic loss rates in twin and singleton pregnancies after transfer of top- versus intermediate-quality embryos. AB - OBJECTIVE: To determine whether embryo quality is associated with early spontaneous loss rates in twin and singleton pregnancies after IVF/intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective, single center analysis. SETTING: The Center of Reproductive Medicine, Arcispedale Santa Maria Nuova, Reggio Emilia, Italy. PATIENT(S): Women undergoing IVF/ICSI and two- or three-embryo transfer of intermediate- and top-quality embryos. INTERVENTION(S): First-trimester sonography at 6 to 7 weeks to determine number of embryos with positive heartbeat. Number of embryos lost was calculated from a second-trimester sonogram. MAIN OUTCOME MEASURE(S): Rates of total pregnancy loss, as related to embryo quality, initial number of embryos, maternal age <35 or > or =35 years, and IVF procedure. RESULTS: A total of 94 losses (23.1% of 407 pregnancies) were counted, with similar proportions in pregnancies after transfer of intermediate- or top-quality embryos. Neither the mode of IVF procedure nor the number of transferred embryos affected the loss rate. In contrast, the loss rate was significantly higher in older mothers after transfer of intermediate-quality embryos (odds ratio [OR 2.4], 95% confidence interval [CI] 1.1-5.5). Losses among singletons were significantly higher compared with losses among twins (OR 2.5, 95% CI 1.1-6.0), but this was observed in top-quality embryos only. CONCLUSION(S): Top-quality but not intermediate-quality ETs are associated with lower early spontaneous loss rates among twin pregnancies after IVF/ICSI. PMID- 16359953 TI - Removal of lysed blastomeres from frozen-thawed embryos improves implantation and pregnancy rates in frozen embryo transfer cycles. AB - OBJECTIVE: To evaluate the effect of degenerated (lysed) blastomere removal on implantation and pregnancy rates in cleavage-stage cryo-embryo transfer (ET) cycles. DESIGN: Randomized clinical trial. SETTING: Private reproductive medical center. PATIENT(S): A total of 88 patients who received frozen-thawed ET, divided into two groups. INTERVENTION(S): Embryo freezing and thawing; opening of the zona pellucida and removal of cryodamaged blastomeres (in the study group), followed by same-day ET. MAIN OUTCOME MEASURE(S): Extent of survival of cleavage stage embryos after the freeze-thaw procedure; embryo implantation and clinical pregnancies. RESULT(S): Oocyte number per patient, fertilization rate, embryo development rate (and quality), and freezing rates were similar in the two groups in the fresh cycle. In the control group, a total of 55 embryos (25%) of the 217 thawed remained fully intact, and 53 (26%) of the 207 in the study group remained intact. The average number of embryos transferred per group was similar (control, 3.4 +/- 0.9; study, 3.3 +/- 0.9). Implantation rates were 12% and 26% in the control and study groups, respectively. The clinical pregnancy rate was 23% in the control group and 64% in the study group when lysed cell removal was performed. CONCLUSION(S): The results show that pregnancy and implantation rates are higher in the study group; therefore, the removal of degenerated blastomeres may be beneficial to all patients who undergo cleavage-stage, frozen-thawed ET. PMID- 16359954 TI - Elective single blastocyst transfer reduces twin rates without compromising pregnancy rates. AB - OBJECTIVE: To compare IVF outcome in single versus two blastocyst transfer in good-prognosis patients. DESIGN: Retrospective data analysis. SETTING: University based IVF practice. PATIENT(S): Assisted reproductive technology outcome was compared in 41 patients who underwent elective single blastocyst transfer and 66 patients who underwent two blastocyst transfer. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Implantation rates, clinical pregnancy rates, cumulative pregnancy rates, twin pregnancy rates. RESULT(S): Blastocyst cycles between January 2003 and August 2004 (n = 238) were reviewed. Criteria for elective single embryo transfer established by our clinic were applied retrospectively to blastocyst cycles. Patients who met the criteria were divided into two groups, those who underwent elective single embryo transfer (eSET) and those who underwent two embryo transfer (2ET). Patient and cycle characteristics were similar for both groups. There were no differences in implantation rate (76% vs. 66%) or pregnancy rate (76% vs. 79%) between the eSET and 2ET groups. There was a significant difference in the number of twin pregnancies (3.2% vs. 62%) in the eSET versus 2ET group. Cumulative pregnancy rates were found to be 83% for both groups. CONCLUSION(S): Single blastocyst transfer can be performed in good prognosis patients without compromising pregnancy rates. Twin pregnancy rates can be significantly reduced with eSET. PMID- 16359955 TI - Elective transfer of single fresh blastocysts and later transfer of cryostored blastocysts reduces the twin pregnancy rate and can improve the in vitro fertilization live birth rate in younger women. AB - OBJECTIVE: To determine the extent to which embryo selection by blastulation and elective single blastocyst transfer, supported by efficient cryostorage of spare embryos, can reduce multiple pregnancies and maintain or improve on the live birth rate from IVF. DESIGN: Prospective, nonrandomized cohort study. SETTING: Sydney IVF, a private clinic in Australia. PATIENT(S): In vitro fertilization patients aged <38 years with three or more usable blastocyst, recruited from April 2000 through December 2001, with pregnancies followed up until August 2004. INTERVENTION(S): Blastocysts were cultured and cryostored with stage-specific culture medium and low oxygen conditions. MAIN OUTCOME MEASURE(S): Fetal heart positive twin pregnancy rate and accumulating "take-home baby" rate per retrieval leading to blastocyst transfer. RESULT(S): Among 121 women who elected single fresh blastocyst transfer (but who could elect to have two frozen blastocysts transferred at once), 79 (65.3%) took home a baby, with a twin pregnancy rate of 7%. Among 285 women who chose two blastocysts for fresh transfer, 184 (64.2%) took home at least one baby, with a twin pregnancy rate of 34% and five perinatal deaths. CONCLUSION(S): With technically appropriate blastocyst culture and freezing, and elective single blastocyst transfer in the fresh cycle, the overall multiple pregnancy rate can be reduced by >75%, permitting in this series a slight increase in the chance of taking home a baby. PMID- 16359957 TI - Optimum number of embryos to transfer in women more than 40 years of age undergoing treatment with assisted reproductive technologies. AB - OBJECTIVE: To determine whether increasing the number of embryos transferred beyond five increases pregnancy rates in women aged > 40 years. DESIGN: Retrospective analysis of cycles performed between January 1998 and July 2003. SETTING: University-affiliated teaching hospital. PATIENT(S): Women aged > 40 years undergoing a fresh cycle with a day-3 ET (n = 863). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy, chemical pregnancy, miscarriage rates, number of viable fetuses at 12 weeks' gestation, live birth rates, and number of babies delivered. RESULT(S): Compared with patients with fewer than five embryos transferred, those having five or more embryos transferred had significantly increased pregnancy rates and live birth rates, more viable fetuses at 12 weeks, and significantly decreased miscarriage rates. None of these outcome variables differed between the five-embryo and more-than-five-embryo groups. There were no differences in outcome when only five embryos were transferred, regardless of whether five or more than five embryos were available. The number of embryos transferred did not significantly influence multiple birth rates. CONCLUSION(S): The present study demonstrates that in women aged > 40 years, five embryos is the optimum number to transfer, and transferring more than five does not confer any additional benefit to clinical outcome. PMID- 16359956 TI - Pregnancies and live births after trophectoderm biopsy and preimplantation genetic testing of human blastocysts. AB - OBJECTIVE: To compare multiple-cell trophectoderm biopsy for preimplantation genetic diagnosis (PGD) from day-5 blastocysts with previously published experience with day-3 cleavage-stage embryos. DESIGN: Retrospective review of laboratory and clinical experience. SETTING: Sydney IVF, a private clinic in Australia. PATIENT(S): Preimplantation genetic diagnosis (PGD) patients age < 44 years with at least one IVF blastocyst suitable for biopsy, recruited from January 2002 through August 2004. INTERVENTION(S): Biopsy of trophectoderm from blastocysts on day 5 or 6, with same-day PGD for mutation testing, translocation testing, aneuploidy screening or sex selection. Spare, normal biopsied blastocysts were cryostored for possible later transfer. MAIN OUTCOME MEASURE(S): Fetal heart-positive pregnancy rate and accumulating live birth rate after adding results from biopsied fresh and frozen blastocysts for particular couples. RESULT(S): In 231 started PGD treatment cycles, unambiguous results were obtained from 974 of 1,050 biopsied blastocysts (93%); all blastocysts survived the biopsy procedure by reconstitution of their blastocele. One hundred nineteen women (median age, 36 years) have had 127 blastocysts transferred fresh (fetal heart positive implantation rate, 41%). Of 146 blastocysts cryostored, 27 have been thawed (all with > 50% cell survival) and 24 transferred (implantation rate, 26%). To date, 53 pregnancies have been delivered or are ongoing, with an additional 4 clinical miscarriages (7%) and 6 subclinical miscarriages (total miscarriage rate, including biochemical pregnancies, 16%). There were no twin pregnancies. CONCLUSION(S): With technically appropriate blastocyst culture and freezing, blastocyst biopsy and cryostorage and later transfer of biopsied blastocysts is shown to be a practical and probably preferable path to preimplantation genetic testing of embryos compared with cleavage-stage embryo biopsy, being accompanied by a high implantation rate (and hence more conducive to elective single embryo transfer) and by a low rate of twinning and miscarriage. PMID- 16359958 TI - Oocytes in the empty follicle: a controversial syndrome. AB - OBJECTIVE: To assess the prevalence and etiology of the empty follicle syndrome (EFS). DESIGN: Observational longitudinal study. SETTING: Tertiary fertility centers. PATIENT(S): All patients beginning in vitro fertilization (IVF) treatment from December 2002 to November 2004 were included. Couples undergoing IVF with donor oocytes or participating in an experimental IVF study were excluded from analysis. INTERVENTION(S): Identification of EFS cycles. Comparing ovarian hyperstimulation strategy, follicle count, and timing of human chorionic gonadotropin (hCG) for final oocyte maturation of the EFS cycles with normal IVF cycles. MAIN OUTCOME MEASURE(S): Number of follicles punctured, number of oocytes recovered, previous and future IVF attempts, and serum hormone levels. RESULT(S): Twenty-five of a total of 1,849 patients were identified with an EFS cycle. Reasons for occurrence of EFS cycles were mistiming of hCG for final oocyte maturation, premature ovulation, and poor ovarian response. None of the affected patients had experienced EFS cycles in earlier IVF attempts nor were there any recurrence in subsequent treatments. CONCLUSION(S): Accurate timing of induction of final oocyte maturation, properly scheduled ovarian hyperstimulation, instruction of patients and doctors, and full workup for IVF are essential for the successful recovery of oocytes. Occurrence of EFS in IVF can normally be attributed to a failure of at least one of these factors and probably rarely or never occurs otherwise. PMID- 16359959 TI - Reproductive outcomes in men with prenatal exposure to diethylstilbestrol. AB - OBJECTIVE: To examine prenatal diethylstilbestrol (DES) exposure in relation to male reproductive outcomes. DESIGN: Prospective observational study. SETTING: Participants were identified through record review, clinical trial participation, or an obstetrics clinic. PATIENT(S): A total of 1,085 DES-exposed and 1,047 unexposed men. INTERVENTION(S): Participants were exposed prenatally to DES through the mother's obstetrics care or clinical trial participation. MAIN OUTCOME MEASURE(S): Infertility; never fathering a pregnancy or live birth; number of pregnancies or live births fathered. RESULT(S): We found little evidence that prenatal DES exposure affects the likelihood of never fathering a pregnancy or live birth, or influences the mean number of fathered pregnancies or live births. Our data suggest that DES-exposed men are slightly more likely to experience infertility (relative risk [RR] = 1.3, 95% confidence interval [CI] = 1.0-1.6). The DES dose and gestational timing did not influence infertility or the number of pregnancies or live births fathered, but results were inconsistent for dose effects on the likelihood of never fathering a pregnancy or a live birth. CONCLUSION(S): Prenatal DES exposure may be associated with a slightly increased risk of having an infertility experience, but does not increase the likelihood of never fathering a pregnancy or a live birth, or the number of pregnancies or live births fathered. PMID- 16359960 TI - Constant decline in sperm concentration in infertile males in an urban population: experience over 18 years. AB - OBJECTIVE: To investigate semen quality in infertile men from an urban population over an 18-year period. DESIGN: Retrospective study. SETTING: Andrology clinic at a city university hospital. PATIENTS: A total of 9,327 men were referred to the clinic for infertility investigation. After excluding those with azoospermia, 7,780 samples were evaluated. INTERVENTION(S): Semen samples were analyzed within half an hour of production using computer-automated semen analysis and phase microscopy. MAIN OUTCOME MEASURE(S): Sperm concentration, morphology, and motility; semen pH and leukocyte concentration; and patient age. RESULT(S): The median patient age over the study period was 31.6 years, with an increase from 30.8 years in 1986 to 34.4 years in 2003. The median sperm concentration was 10.25 million/mL for the entire period, with a decline from 27.75 million/mL in 1986 to 4.60 million/mL in 2003. The median proportion of normally shaped spermatozoa was 15%, and the proportion of motile spermatozoa was 21%. The pH increased from 7.4 in 1986 to 7.9 in 2003, and the median leukocyte concentration was 1.50 million/mL. CONCLUSION(S): A constant decline in median sperm concentration was found in the infertile men. The reason for this remains unclear, although the high pH and leukocyte concentration indicates the involvement of infection. PMID- 16359961 TI - Age as a limiting factor for successful sperm retrieval in patients with nonmosaic Klinefelter's syndrome. AB - OBJECTIVE: To determine factors affecting successful sperm retrieval by testicular sperm extraction in patients with nonmosaic Klinefelter's syndrome. DESIGN: Medical record analysis for nonmosaic Klinefelter's syndrome patients who underwent testicular sperm extraction. SETTING: Three university-based tertiary centers. PATIENT(S): Fifty-one patients with nonobstructive azoospermia related to nonmosaic Klinefelter's syndrome. INTERVENTION(S): Testicular sperm extraction. MAIN OUTCOME MEASURE(S): Correlation of patient characteristics; serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (T); as well as testicular volume with success in testicular sperm extraction. RESULT(S): We succeeded in obtaining spermatozoa in 26 patients and failed in 25. Levels of LH, FSH, and T and testicular volume did not differ between patient groups defined by success and failure. Median ages for successful and failed testicular sperm extraction were 31 and 38 years, respectively (statistically significant difference). When we analyzed success rates of testicular sperm extraction between two patient groups (<35 and > or =35 years old), the percentage of successful recovery of spermatozoa decreased after the age of 35 years (statistically significant difference). CONCLUSION(S): Testicular sperm extraction should be recommended before the critical age of 35 years. PMID- 16359962 TI - Intracytoplasmic sperm injection: a novel selection method for sperm with normal frequency of chromosomal aneuploidies. AB - OBJECTIVE: To test a newly invented intracytoplasmic sperm injection (ICSI) sperm selection method based on sperm hyaluronic acid (HA) binding. DESIGN: Comparison of chromosomal disomy and diploidy frequencies in sperm arising from semen and in HA-bound sperm. SETTING: Academic andrology laboratory. PATIENT(S): Men presenting for semen analysis. INTERVENTION(S): Washed sperm fractions of 32 semen samples were applied to Petri dishes or glass slides coated with immobilized HA. The unbound sperm were rinsed gently, and the HA-bound sperm were removed with an ICSI pipette. The control sperm population was the unselected sperm. Both HA-selected and unselected sperm were treated with fluorescence in situ hybridization with centromeric probes for the X, Y, and 17 chromosomes. MAIN OUTCOME MEASURE(S): Chromosomal disomy and diploidy frequencies. RESULT(S): In the HA-bound sperm (495-2,079 per man, 41,670 in all) compared with unselected sperm (4,770 per man, 162,210 in all), the chromosomal disomy frequencies were reduced to 0.16% from 0.52%, diploidy to 0.09% from 0.51%, and sex chromosome disomy to 0.05% from 0.27% (a 5.4-fold reduction vs. 4-fold respective increase in ICSI offspring). CONCLUSION(S): The HA sperm selection method for ICSI, which is based on a relationship between sperm receptors for zona pellucida and HA, will likely reduce the potential genetic complications and adverse public health effects of ICSI. PMID- 16359963 TI - Genome-wide linkage analysis to age at natural menopause in a community-based sample: the Framingham Heart Study. AB - OBJECTIVE: To identify chromosomal regions linked to age at natural menopause. DESIGN: Two-generation families studied with a 10-centimorgan (cM) genome-wide scan. SETTING: The Framingham Study, a community-based epidemiologic study. PATIENT(S): A total of 861 women in 291 families reporting a natural menopause (mean age at menopause, 49 years). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Multipoint variance components linkage analysis was performed with Genehunter software (Whitehead Institute, Massachusetts Institute of Technology, Boston, MA) for age at natural menopause. RESULT(S): In the crude variance components analysis, 11 chromosomes had a log odds ratio (LOD) score of > or =1.0. Two chromosomal regions revealed suggestive linkage: chromosome 8 at 26 cM (LOD 2.6; nearest marker GATA23D06) and chromosome 16 at 11 cM (LOD 2.4; nearest marker ATA41E04). In the analysis adjusted for generation, smoking, and body mass index, chromosome 11 revealed suggestive linkage at 113 cM (LOD 2.1; nearest marker GATA23E06). CONCLUSION(S): We have identified novel loci suggestive for linkage to age at natural menopause. Further research is needed to identify genes involved in the onset of menopause, which might provide insights into loss of fertility. PMID- 16359964 TI - Procoagulant state after raloxifene therapy in postmenopausal women. AB - OBJECTIVE: To investigate the effects of raloxifene on the hemostatic system in postmenopausal women. DESIGN: A prospective longitudinal study. SETTING: Outpatient clinic of the Faculty of Medicine of Ribeirao Preto, Brazil. PATIENT(S): Sixteen postmenopausal women aged 56.8 +/- 5.9 years (mean +/- SD). INTERVENTION(S): Raloxifene hydrochloride (60 mg once daily) was administered orally for a period of 6 months. MAIN OUTCOME MEASURE(S): Plasma activities of coagulation factors (II, V, VII, VIII, IX, X, XI, XII, and fibrinogen), prothrombin-derived fragment 1+2, and activated protein C (APC) sensitivity ratio were measured at baseline and after 1, 3, and 6 months of treatment. RESULT(S): Factor VIII activity increased by 17.1% and 26.9% at 3 and 6 months of treatment, respectively, compared with baseline. Factor XI and FXII activities significantly increased by 10.9% and 43.1%, respectively, after 6 months compared with baseline. A significant reduction of APC sensitivity ratio also was observed after 6 months of treatment. CONCLUSION(S): A procoagulant state characterized by increased factor VIII, XI, and XII plasma levels and by reduced APC sensitivity was observed after raloxifene therapy in post-menopausal women. PMID- 16359965 TI - Mullerian-inhibiting substance reflects ovarian findings in women with polycystic ovary syndrome better than does inhibin B. AB - OBJECTIVE: To investigate Mullerian-inhibiting substance (MIS) levels in women with polycystic ovary syndrome (PCOS), as well as relationships to ovarian morphology, levels of inhibin B, and other reproductive hormones. DESIGN: Prospective clinical study. SETTING: Academic endocrinology centers in Palermo, Italy and New York. PATIENT(S): Forty-six women with PCOS, recruited on the basis of the classic criteria of chronic anovulation and hyperandrogenism, and 25 age matched ovulatory controls. INTERVENTION(S): Fasting blood was obtained in all subjects in the early follicular phase (days 5-6) after spontaneous or induced menses (in PCOS), and transvaginal ultrasounds were performed. MAIN OUTCOME MEASURE(S): Assessment of values for luteinizing hormone (LH), testosterone (T), androstenedione (A), estradiol (E2), inhibin B, MIS, fasting insulin, and the calculated quantitative sensitivity check index (QUICKI), as well as assessments of ovarian volume and blood flow. RESULT(S): Women with PCOS had higher LH, T, and A; higher insulin and lower QUICKI; and higher ovarian volume and lower pulsatility index. Inhibin B concentrations were statistically significantly higher in PCOS patients (70 +/- 8.0 vs. 40 +/- 3.4 pg/mL), as was MIS (6.7 +/- 0.9 vs. 4.6 +/- 0.5 ng/mL). Inhibin B had a statistically significant direct correlation with levels of MIS (r = 0.351). However, MIS, but not inhibin B, had a statistically significant positive correlation with ovarian size (r = 0.350); the reproductive hormones LH, T, A, and E2; and insulin (r = 0.249), independent of body mass index. Women with PCOS with the highest levels of MIS had higher ovarian volumes and values of LH, T, A, and insulin. CONCLUSION(S): Measurements of MIS reflect ovarian findings in PCOS better than levels of inhibin B and are more frequently elevated. However, MIS lacks sensitivity for use as a diagnostic tool in PCOS. PMID- 16359966 TI - Anatomical evidence for in utero androgen exposure in women with polycystic ovary syndrome. AB - OBJECTIVE: To determine whether women with polycystic ovary syndrome (PCOS) have masculinized finger length patterns compared to women without PCOS. DESIGN: A case control study. SETTING: University teaching hospital and in vitro fertilization unit. PATIENT(S): Seventy women aged between 18 and 40 years with PCOS were compared to 70 women without PCOS. INTERVENTION(S): Measurement of the second to fourth finger length ratio on the ventral surface of the left and right hand from the basal crease of the digit to the tip was made using Vernier calipers. MAIN OUTCOME MEASURE(S): The second to fourth finger length ratio. RESULT(S): We found a significantly reduced ratio in the right hand of the women with PCOS compared to the controls. The geometric mean right finger length ratio in the PCOS group was 98.3% that of the controls (95% confidence interval, 99.3% 97.3%). CONCLUSION(S): Here we show a subtle difference in the finger length pattern of women with PCOS. This may constitute anatomical evidence of in utero androgen exposure in PCOS. PMID- 16359967 TI - Comparison of the clinical efficacy of flutamide and spironolactone plus Diane 35 in the treatment of idiopathic hirsutism: a randomized controlled study. AB - OBJECTIVE: To compare the effectiveness of flutamide and spironolactone plus Diane 35 in the treatment of idiopathic hirsutism (IH). DESIGN: Prospective randomized clinical study. SETTING: Social Security Agency, Aegean Obstetrics and Gynecology Teaching Hospital, Department of Reproductive Endocrinology and Infertility, a tertiary referral center. PATIENT(S): Eighty women with IH who had been admitted to our center. INTERVENTION(S): Patients were randomly and equally assigned to receive flutamide (250 mg/d for the first 10 days of the cycle) and spironolactone plus Diane 35 (spironolactone 100 mg/d; Diane 35 is a combination tablet of 2 mg cyproterone acetate and 35 microg ethinylestradiol/d) for 9 months. MAIN OUTCOME MEASURE(S): Modified Ferriman-Gallwey (F-G) score and hormonal profile before treatment and at the end of treatment. RESULT(S): In spite of the fact that there were no statistically significant differences in modified F-G scores in between the two groups, a significant decrease in modified F-G scores was observed in the flutamide (from 19.93 +/- 4.31 to 15.58 +/- 4.28) and spironolactone plus Diane 35 (from 18.77 +/- 3.76 to 14.54 +/- 3.29) groups. There were also no differences between the two groups with respect to hormonal profiles including serum FSH, LH, free T, 17-OH-progesterone, and DHEAS levels. No major side effect, including liver toxicity, was observed in either group. CONCLUSION(S): Our data suggest that both therapies have similar effectiveness in the treatment of IH. PMID- 16359968 TI - Measurement of total testosterone in women: comparison of a direct radioimmunoassay versus radioimmunoassay after organic solvent extraction and celite column partition chromatography. AB - OBJECTIVE: To evaluate the clinical usefulness for the measurement of testosterone (T) values in the "low" female range with a direct radioimmunoassay (RIA) for total T by comparing total T values measured by this assay with values determined by conventional RIA after organic solvent extraction/column chromatography. DESIGN: Cross-sectional study. SETTING: Victoria, Australia. PATIENT(S): Two hundred fifty-nine healthy women, aged 18-75 years, recruited from the community. INTERVENTION(S): Fasting serum samples were obtained and stored at -80 degrees C. MAIN OUTCOME MEASURE(S): Total T measurement. Total T was measured by the direct RIA method using antibody-coated tubes and iodine labeled T tracer. For comparison, total T levels were also measured using the conventional RIA method after organic solvent (ethyl acetate:hexane [3:2]) extraction and celite column partition chromatography before RIA. RESULT(S): The mean T level by direct RIA was 0.76 nmol/L (median, 0.70; SD, 0.54; minimum, 0.10; maximum, 3.2). The mean difference between the two measurements (direct RIA - conventional RIA) was -0.28 (SD, 0.3). The direct RIA value was 63% (95% confidence interval, 26%-155%) of the conventional RIA estimate. For the classification of values at the lower end of the range, there was very good agreement beyond chance (kappa = 0.68-0.74) for values in approximately the lowest 10th-40th percentiles. CONCLUSION(S): The direct RIA is a clinically useful assay for the study of the issue of "low" T within the female population. PMID- 16359969 TI - Changes in the T-helper cytokine profile and in lymphocyte activation at the systemic and local levels in women with endometriosis. AB - OBJECTIVE: To estimate regulatory cytokine synthesis and lymphocyte activation in the peripheral blood and endometrial tissue of patients with endometriosis. DESIGN: Controlled clinical study. SETTING: Medical center. PATIENT(S): Fifteen women with laparoscopically diagnosed endometriosis; 20 gynecologically healthy women with previously documented fertility (control group). INTERVENTION(S): Peripheral venous blood sampling; laparoscopic collection of ectopic and matched eutopic endometrium. MAIN OUTCOME MEASURE(S): Messenger RNA (mRNA) for interleukin (IL)-2, IL-4, and IL-10 expression in peripheral and endometrium lymphocytes was assessed by real-time reverse transcriptase polymerase chain reaction; intracellular synthesis of these cytokines and lymphocyte phenotype profile were established by flow cytometry. RESULT(S): Both mRNA expression and intracellular synthesis of IL-4 and IL-10 were sharply increased in peripheral lymphocytes. The same changes were observed for IL-4 in ectopic endometrium of women with endometriosis. Simultaneously, elevation of the amount of pan-B cells, CD20+CD5+ B-1 cells, and activated HLA-DR+CD20+ B lymphocytes was observed in endometriosis lesions. Only an enhanced amount of B lymphocytes was seen in eutopic endometrium. CONCLUSION(S): Endometriosis development is accompanied by the activation of a T-helper type 2 immune response at the systemic and local levels. Our results support the hypothesis regarding the autoimmune nature of endometriosis and can explain the high level of autoantibody production in patients with endometriosis. PMID- 16359970 TI - Quantification of mitochondrial DNA using real-time polymerase chain reaction in patients with premature ovarian failure. AB - OBJECTIVE: To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group. DESIGN: Prospective study. SETTING: Genome Research Center for Reproductive Medicine and Infertility, Korea Ministry of Health & Welfare. PATIENT(S): Thrity patients with POF and 30 control individuals. INTERVENTION(S): The mitochondrial DNA content was quantified using real-time PCR. The effectiveness of the assay was determined by relative quantification using the comparative threshold cycle (CT) method. MAIN OUTCOME MEASURE(S): Relative quantification of mitochondrial DNA content. RESULT(S): The mitochondrial DNA content was significantly lower in the POF group than in the control group (0.58 +/- 0.38 vs. 1.15 +/- 0.67; P < .01). In both groups, there was a significant positive correlation between the mitochondrial DNA/28S rRNA ratio and mitochondrial DNA CT (control group: r = 0.774; P < .001; POF group: r = 0.556; P = .001) and a significant negative correlation between the mitochondrial DNA/28S rRNA ratio and 28S rRNA CT (control group: r = -0.677; P < .001; POF group: r = -0.627; P = .001). CONCLUSION(S): This study has established the clinical feasibility of quantifying amounts of mitochondrial DNA, relative to an internal standard, using real-time PCR. Further studies are warranted to elucidate the roles of apoptosis and mitochondrial function in the pathogenesis of POF. PMID- 16359971 TI - Management of patients receiving long-term treatment with mifepristone. AB - OBJECTIVE: To determine clinical side effects and biochemical and hematological abnormalities in patients with nonresectable meningioma on long-term mifepristone (RU 486) therapy. DESIGN: Long-term mifepristone administration in patients with meningioma. SETTING: Outpatient clinic of a university hospital. PATIENT(S): Sixteen women and 9 men aged 22-80 years with nonresectable meningioma. INTERVENTION(S): Mifepristone (200 mg daily). One patient received treatment for more than 13 years; six received treatment for 10-12 years; five received treatment for 4-9 years; eight received treatment for 1-4 years; and the remainder received treatment for 4-10 months. MAIN OUTCOME MEASURE(S): Evaluation of side effects and of hematological and biochemical abnormalities. RESULT(S): Fatigue was observed in 22 of 25 patients. Endometrial hyperplasia occurred in one premenopausal woman and one postmenopausal woman. Another two women had endometrial thickening without hyperplasia. There were no consistent abnormalities in liver or renal function or in any other biochemical or hematological parameters. One subject (on long-term dexamethasone) developed hypoadrenalism, which responded to treatment. CONCLUSION(S): Mifepristone can be administered for prolonged periods. Ultrasound should be performed if irregular vaginal bleeding occurs. In asymptomatic women, it should be performed annually. If endometrial thickening is observed, then endometrial biopsy is recommended. Because biochemical hypothyroidism has been reported during long-term mifepristone therapy, thyroid function tests should be performed annually. PMID- 16359972 TI - Modes of conception and multiple pregnancy: a national survey of babies born during one week in 2003 in the United Kingdom. AB - OBJECTIVE: To collect data on every baby born in one week in the United Kingdom with respect to mode of conception, multiplicity of pregnancy, and outcome. DESIGN: A questionnaire completed by the midwife birth attendant. SETTING: All maternity units in the United Kingdom. PATIENT(S): All mothers delivering during the week of April 6-12, 2003. INTERVENTION(S): Questionnaire survey. MAIN OUTCOME MEASURE(S): Mode of conception (both spontaneous and assisted), type of fertility treatment, multiplicity of pregnancy, mode of delivery, gestational age, and fetal outcome. RESULT(S): Data were received from 178 maternity units (72.7%) on 6,913 deliveries: 6,812 (98.54%) were singleton, 100 (1.45%) twin, and 1 (0.01%) triplet. In total, 7,015 babies were born. Of all pregnancies, 6,638 (96%) (including the only triplet) were conceived spontaneously and 133 (1.9%) with assistance. The multiple pregnancy rate was significantly greater in assisted (13.5%) than in spontaneous (1.2%) conceptions. Of the multiple pregnancies after fertility treatment, 16.7% resulted from clomifene citrate therapy, 72.2% from IVF or frozen embryo replacements (FET), and 5.6% from superovulation with intrauterine insemination (IUI). The multiple pregnancy rate after IVF/FET (26%) was significantly higher than the one after clomifene citrate therapy (7.3%). In total, 41 babies were born after clomifene citrate therapy, 50 after IVF/ET, and eight after superovulation IUI. The live birth rate was higher for singleton (98.2%) than multiple pregnancies (93.6%). CONCLUSION(S): Multiple pregnancy from IVF and related treatments remains a significant problem and contributes a greater burden than ovulation induction therapies. PMID- 16359973 TI - Follow-up of successful bilateral placement of Essure microinserts with ultrasound. AB - OBJECTIVE: To evaluate the reliability of pelvic X-ray and transvaginal ultrasound to localize Essure microinserts (Conceptus, San Carlos, California) after successful placement in both fallopian tubes 3 months after placement. DESIGN: Prospective, observational study. SETTING: Gynecology departments at two teaching hospitals. PATIENT(S): One hundred eighty-two patients who underwent hysteroscopic sterilization by placement of Essure microinserts between August 2002 and August 2004. INTERVENTION(S): Transvaginal ultrasound, pelvic X-ray, and hysterosalpingography (HSG) 3 months after sterilization with Essure. MAIN OUTCOME MEASURE(S): Transvaginal ultrasound confirmation of correct localization of microinserts after a 3-month follow-up. RESULT(S): In 150 of 182 patients, confirmation of successful bilateral placement of two microinserts (300 devices) was possible. In 9 patients it was not possible to identify both devices with ultrasound, or there was doubt about the extension of the device through the uterotubal junction. The other 291 devices were identified as being in a good position. CONCLUSION(S): Hysterosalpingography at the 3-month follow-up after successful placement of Essure microinserts can be replaced by transvaginal ultrasonography. A 3-month follow-up with HSG after the Essure procedure is only required after unsatisfactory placements. In those patients in whom transvaginal ultrasonography cannot confirm satisfactory localization, a complementary pelvic X-ray should be performed. PMID- 16359974 TI - Use of a zona laser treatment system in assisted hatching: optimal laser utilization parameters. AB - OBJECTIVE: This study aimed to establish the benefit of laser micromanipulation of the zona pellucida as a method of assisted hatching and to determine the optimal technical settings. DESIGN: Prospective pilot study. SETTING: University teaching hospital. ANIMAL(S): Six- to 8-week-old F1 female mice. INTERVENTION(S): Mouse embryos were treated with a laser (Hamilton Thorne Research) using both research and preset clinical settings. The research option allows the user to alter pulse duration, number of pulses, and beam intensity. In this study, laser intensity and duration, as well as the depth and number of disruptions, were altered. MAIN OUTCOME MEASURE(S): Embryos were scored after treatment as either arrested blastocysts, arrested during hatching, hatched, or degenerate. RESULT(S): The treatment where the laser intensity was halved but the number of laser pulses used was doubled was the most beneficial treatment. Zona thinning was not effective. CONCLUSION(S): The study showed that laser-assisted hatching can safely and significantly increase the percentage of embryos reaching the hatched blastocyst stage. A proportionately sized hole in a human embryo may have the same beneficial effect as has been shown in this study. PMID- 16359975 TI - Radiologic and surgical demonstration of uterine polyposis. AB - We describe a rare case of uterine polyposis that was visualized by sonohysterogram, hysterosalpingogram, and hysteroscopy. PMID- 16359976 TI - Mosaic ring Y chromosome in two normal healthy men with azoospermia. AB - OBJECTIVE: To better define by molecular and cytogenetic techniques ring Y chromosomes detected in 2 infertile men. DESIGN: Case report. SETTING: Molecular genetics/cytogenetics unit in a university hospital. PATIENT(S): Two infertile men with azoospermia, presenting a normal male phenotype with complete masculinization. INTERVENTION(S): Karyotype and genetic counseling. MAIN OUTCOME MEASURE(S): Metaphases were studied by standard G- and Q-banding; fluorescent in situ hybridization and PCR were performed to analyze specific Y chromosome regions. RESULT(S): Chromosomal analysis detected a mosaicism with a Y chromosome ring cell line in 92% (patient 1) and 95% (patient 2) of the metaphases, coexisting with a 45,X cell line in the remaining metaphases. In patient 1, PCR analysis showed the presence of AZFa region and a partial deletion of AZFb region; AZFc region was deleted. In patient 2 all three AZF regions were deleted. CONCLUSION(S): A 45,X/46,X,r(Y) mosaicism can be detected not only in patients with Ullrich-Turner syndrome and in patients with various degrees of genital ambiguity but also in men presenting a normal phenotype. Their azoospermia can be explained by partial or total deletion of AZF regions. PMID- 16359977 TI - Novel presentation of retroperitoneal germ cell tumor with loss of ejaculation but preservation of erection in two patients. AB - OBJECTIVE: To examine the effect of large-volume retroperitoneal germ cell tumor on nerve fibers required for erection and ejaculation. DESIGN: Retrospective case reports. SETTING: Cancer treatment center. PATIENT(S): Two male patients presenting with loss of ejaculation but preservation of erection. RESULT(S): Retroperitoneal lymph nodes lie very close to the sympathetic nerve fibers that control ejaculation. Large-volume tumors would encase the sympathetic plexuses at L1 to L3, making ejaculation impossible. Parasympathetic fibers arise from S2 to S4, so they would not be damaged, preserving erection. CONCLUSION(S): Thorough history taking is important in the investigation of infertility. Presentation with these symptoms requires further investigation at an early stage to exclude retroperitoneal tumor. PMID- 16359978 TI - Reversibility of hypogonadotropic hypogonadism in a patient with the juvenile form of hemochromatosis. AB - OBJECTIVE: To report a case of complete reversibility of hypogonadotropic hypogonadism with intensive venesection treatment in juvenile hemochromatosis. DESIGN: Case report. SETTING: Endocrine department of Hippocrateion Hospital of Athens. PATIENT(S): A 25-year-old man who presented with hypogonadotropic hypogonadism and severe iron overload due to juvenile hemochromatosis and who was initially treated with phlebotomies and androgen substitution. INTERVENTION(S): Intensification of chelation therapy. MAIN OUTCOME MEASURE(S): Clinical evaluation, serum ferritin concentration, and biochemical assessment of pituitary function were performed periodically. RESULT(S): One year after normalization of serum ferritin levels and transferrin saturation was achieved, he became eugonadal. CONCLUSION(S): We believe that hypogonadotropic hypogonadism in juvenile hemochromatosis may be reversible by a consequent venesection therapy probably because treatment was intensive and promptly introduced at a young age. PMID- 16359979 TI - Gadolinium radiologic contrast is a useful alternative for hysterosalpingography in patients with iodine allergy. AB - OBJECTIVE: To report the successful use of gadolinium radiologic contrast for hysterosalpingography. DESIGN: Case report. SETTING: Outpatient care in a private practice setting. PATIENT(S): One female patient entering and finishing study. INTERVENTION(S): Hysterosalpingogram. MAIN OUTCOME MEASURE(S): Interpretability of radiological imaging. RESULT(S): Visualization of a normal uterine cavity and fallopian tubes with bilateral patency was confirmed during the procedure. Radiologic films were created that document the results. The best visualization of the internal pelvic anatomy was noted during the actual procedure. There was a lighter image noted on the films taken from the use of gadolinium rather than the traditional iodinated contrast. CONCLUSION(S): Gadolinium is a safe and useful alternative to iodinated contrast for hysterosalpingography in patients with iodine allergy. PMID- 16359980 TI - Development of severe ovarian hyperstimulation syndrome after inadvertent stimulation with a gonadotropin-releasing hormone agonist and human menopausal gonadotropin in a pre-existing early pregnancy. AB - OBJECTIVE: To report a case of severe ovarian hyperstimulation syndrome (OHSS) after inadvertent GnRH long protocol/hMG stimulation in a pre-existing early pregnancy. DESIGN: Case report. SETTING: Private infertility clinic. PATIENT(S): A 28-year-old woman who conceived spontaneously following IVF and cryo-embryo transfer (cryo-ET). INTERVENTION(S): IVF/intracytoplasmic sperm injection (ICSI), cryo-ET, analgesia, and forced diuresis. MAIN OUTCOME MEASURE(S): Viable pregnancy. RESULT(S): Viable pregnancy with OHSS despite inadvertent administration of GnRH-agonist, stimulation with hMG, and ET in a pre-existing pregnancy. CONCLUSION(S): Observation of follicle development following stimulation during pregnancy with low quantity and poor quality oocytes combined with abnormal endometrium. PMID- 16359981 TI - Contraceptive information after endometrial ablation. AB - We reviewed the records of the patients considered "fertile" in a consecutive series of 206 patients treated by intrauterine balloon ablation for dysfunctional uterine bleeding, and three pregnancies were observed among 58 patients (5.2%), with two spontaneous abortions and a placenta accreta at 26 weeks. These findings lead us to conclude that balloon ablation is not contraceptive and that use of a supplemental contraceptive method should be planned. Hysteroscopic endometrial ablation and nonhysteroscopic endometrial thermal ablation are the first-line conservative surgical treatments for dysfunctional uterine bleeding. Their use reduces the rate of hysterectomies for this common problem. Because of their effects on the endometrium and uterine cavity (synechiae), these treatments are indicated only for patients who do not wish to remain fertile. PMID- 16359982 TI - Influence of embryo quality and number of previous cycles on pregnancy and multiple pregnancy rates in women aged 35 to 37 years who received two or three embryos. AB - Women aged 35-37 years undergoing IVF-ET with fresh embryos at a university infertility center were analyzed to evaluate factors useful in determining whether two or three embryos should be transferred in this age group. Embryo quality and number, but not number of previous failed cycles, were important in determining outcome, and all triplet pregnancies could have been avoided at our program in this age group by limiting the transfer to two good-quality embryos, without reducing pregnancy rates. PMID- 16359983 TI - Effect of prefreeze assisted hatching on postthaw survival of mouse embryos. AB - Embryos that underwent assisted hatching before freezing had a statistically significant higher survival rate and blastocyst formation rate compared with controls. Therefore, breaching the integrity of the zona pellucida and directly exposing the embryos to cryoprotectants not only does not result in damaging the embryos but might also afford them an advantage as they go through the freeze thaw cycle. PMID- 16359984 TI - Effect of chromium supplementation on insulin resistance and ovarian and menstrual cyclicity in women with polycystic ovary syndrome. AB - In women with polycystic ovary syndrome, chromium picolinate (200 microg/d) improves glucose tolerance compared with placebo but does not improve ovulatory frequency or hormonal parameters. This pilot study indicates that future studies in the polycystic ovary syndrome population should examine higher dosages or longer durations of treatment. PMID- 16359985 TI - Oocytes collected 60 hours after human chorionic gonadotropin administration from patients with polycystic ovary syndrome who underwent limited ovarian stimulation resulted in clinical pregnancies and the delivery of a healthy baby. AB - This was a prospective pilot study of 11 polycystic ovary syndrome (PCOS) patients primed with limited ovarian stimulation. Oocytes were collected 60 hours after human chorionic gonadotropin (hCG) and resulted in pregnancies and the delivery of a healthy baby. PMID- 16359986 TI - The gonadotropin-releasing hormone (GnRH)-1 gene, the GnRH receptor gene, and their promoters in patients with idiopathic hypogonadotropic hypogonadism with or without resistance to GnRH action. AB - The study included 26 patients with idiopathic hypogonadotropic hypogonadism (24 sporadic and 2 familial). The Pro146Ser mutation was identified in the gonadotropin-releasing hormone receptor (GnRHR) gene in two sisters as well as in their mother, and one polymorphism in the GnRH1 gene (the Trp16Ser) was identified in four patients. No mutations in transcription factor-binding sites of their promoters were identified. Three patients (one male and two sisters) were found with resistance to GnRH action. No mutations were identified in the male, whereas in the females the mutation Pro146Ser in the GnRHR was identified in heterozygosity. PMID- 16359987 TI - Plasma matrix metalloproteinase activity during the menstrual cycle of healthy ovulatory patients who underwent tubal ligation. AB - The plasma profiles of active metalloproteinase (MMP)-1, MMP-2, and MMP-9 levels during the normal menstrual cycle in ovulating, healthy and tubal ligated patients were studied. The active MMP-1 and MMP-9 plasma levels were found to vary in the menstrual cycle. Active MMP-2 levels, however, are constant during the menstrual cycle. PMID- 16359988 TI - Sex steroid milieu does not alter the impact of fasting on leptin levels in women. AB - The impact of stressors upon some metabolic and endocrine parameters differs when the gonad is present and in the follicular vs. luteal phase. However, a high vs. low sex steroid milieu did not alter the metabolic response, as assessed by leptin levels, in response to the stressor of fasting. PMID- 16359989 TI - Laparoscopic excision of ovarian cysts does not result in antiovarian humoral autoimmunity. AB - To determine whether the ovarian trauma consequent to the laparoscopic removal of a cyst could result in the development of a humoral immunity, antiovarian antibodies were assayed in serum samples obtained from 40 women before and after cystectomy. PMID- 16359990 TI - Vascular endothelial growth factor plays more than an angiogenic role in the female reproductive system. AB - Although the most well-known function of vascular endothelial growth factor is its angiogenic activity, it plays more than a simple angiogenic role in the female reproductive system as it is involved in a number of key events in the course of the ovulatory cycle. The modulation of vascular endothelial growth factor expression varies in different reproductive tissues, probably related to its various functions at these different sites. PMID- 16359991 TI - Leptin and its receptors in human fetal and adult ovaries. AB - The objective of the study was to investigate the immunocytochemical expression and presence of messenger RNA (mRNA) transcripts for leptin and its receptors in ovaries from human adults and adolescents and second- and third-trimester fetuses. Staining for leptin and the long form of its receptor was identified in oocytes of follicles from primordial stages onward, and for leptin only in granulosa cells of a minority of the samples. Expression of mRNA transcripts for both ligands was detected in all the samples tested. PMID- 16359992 TI - Optimizing assisted reproduction: impact of low-dose gonadotropin-releasing hormone agonist on in vitro fertilization outcome. AB - A novel method of assessing factors that could benefit in vitro fertilization outcome is illustrated by comparison of a low (250 microg) with a standard (500 microg) dose of gonadotropin-releasing hormone agonist, revealing a small decrease in cancellation rates. PMID- 16359993 TI - Staff management in the in vitro fertilization laboratory. AB - Although many IVF centers have implemented a Quality Management System, staff management in the IVF laboratory is still highly neglected. This article describes methods for staff development and monitoring of staff performance in the IVF laboratory. PMID- 16359994 TI - Therapeutic dosages of raloxifene do not modify myeloperoxidase and F2alpha isoprostane levels in postmenopausal women. AB - We investigated the effect of a therapeutic dose of raloxifene on the plasma levels of myeloperoxidase and F2alpha-isoprostanes, two markers of oxidative stress recently described as reliable indicators of coronary heart disease. Contrary to changes described in the literature for estrogens (E), raloxifene did not modify the levels of either myeloproxidase or F2alpha-isoprostanes after 3 or 6 months of treatment. PMID- 16359995 TI - Effect of monocyte chemoattractant protein-1 and estradiol on the secretion of vascular endothelial growth factor in endometrial stromal cells in vitro. AB - To explore the initial activity of endometrial stromal cells (ESCs) and the participation of monocyte chemoattractant protein-1 (MCP-1) in the histogenesis of endometriosis, vascular endothelial growth factor (VEGF) secretion of ESCs and the effect of MCP-1 on VEGF secretion of ESCs were observed in vitro. The VEGF level was detected in ESC culture media and was increased significantly when E2 or MCP-1 was added to the media, especially in the presence of E2 plus MCP-1. The VEGF secretion was higher in the ESCs of women with endometriosis than in those women without endometriosis. PMID- 16359996 TI - Predictive value of poor ovarian response to initial gonadotropin stimulation. PMID- 16359998 TI - The physical properties of seminal fluid-reproducibility and intrasubject variation? PMID- 16360000 TI - Two sources of error in pop-out localization. AB - An odd-one-out stimulus, such as a vertical bar among horizontals, pops out from the background and is easily detected, but its location may be slightly ambiguous. Four observers were asked to pinpoint these stimuli on thousands of trials, in 5 x 5 and 9 x 9 arrays of Gabor patches. We found they made frequent errors toward neighbors of the target. Over a range of performance from 41% to 96% correct, the frequency of neighbor errors was well described by a linear function of the total error frequency, a function that might result from mixing together two spatial distributions--one broad, the other narrow. We suggest that these represent two sources of error in pop-out localization; one might correspond to a higher visual area with imprecise retinotopic mapping, and the other to a more fine-grained localization process in primary visual cortex. PMID- 16360002 TI - Introduction. PMID- 16360001 TI - Sensitivity to spatial and temporal modulations of first-order and second-order motion. AB - This study characterises the spatiotemporal "window of visibility" for first order motion (luminance-modulated noise) and three varieties of second-order motion (contrast-modulated, polarity-modulated and spatial length-modulated noise). Direction-identification thresholds (minimum modulation depth producing 79.4% correct) were measured for each motion pattern (acuity permitting) over a five octave range of spatial and temporal frequencies (0.5-16 c/deg and 0.5-16 Hz respectively). Thresholds were converted into modulation sensitivity (1/threshold). For first-order motion patterns, sensitivity functions were generally bandpass. However, for second-order motion patterns, functions were predominantly lowpass in nature. In particular, the functions corresponding to contrast-modulated and polarity-modulated noise were virtually identical in terms of shape and sensitivity. However, sensitivity to modulations of spatial length was extremely poor and more lowpass, suggesting that additional strategies, perhaps a feature-based system, may be required for encoding motion of images of this type. PMID- 16360003 TI - Molecular mechanisms in cancer: what should clinicians know? AB - Normal cells are influenced by a variety of environmental and host influences that can produce pro-carcinogenic mutations. Either a single or a series of mutations might result in cellular transformation. Like normal cells, most cancer cells use multiple redundant intracellular signaling pathways to ensure the maintenance and viability of functions critical to their survival. Thus, cellular pathways that are integral to cell function, survival, proliferation, and receptor expression are potential targets for therapeutic intervention. One example of this is the epidermal growth factor receptor signaling pathway. Other potential targets are molecules that mediate processes through which tumors produce angiogenic and invasion factors that stimulate host blood vessel growth into tumors and allow tumor growth and metastasis, such as the vascular endothelial growth factor. Targeting of downstream events that result in cellular apoptosis is another potential strategy. Continued investigations may result in the development of proteomic profiling databases through which a patient might be matched with molecular signatures in a library and upon which individualized cancer therapies might be selected. In this way, clinicians might recommend combinations of molecularly targeted agents and other therapies on the basis of an individual patient's proteomic profile. PMID- 16360004 TI - Brief communication: a new combination in the treatment of advanced pancreatic cancer. AB - Single-agent gemcitabine is currently the standard treatment for advanced pancreatic cancer. A majority of recent phase III trials have shown either marginal or no improvement in survival using gemcitabine in combination with other chemotherapeutic and novel agents. Our group recently reported that the combination of gemcitabine with erlotinib was associated with a significant prolongation of survival and improvement in progression-free survival in advanced pancreatic cancer. PMID- 16360005 TI - Molecular mechanisms and targeting of colorectal cancer. AB - Targeted therapies that are approved for metastatic colorectal cancer are divided into two groups: those affecting vascular endothelial growth factor (VEGF) known to interrupt tumor growth and metastasis (also called neo-angiogenesis), and agents that affect the tumor directly by interrupting the epidermal growth factor (EGF) and its receptor. Anti-angiogenic VEGF therapies are divided into two categories: one affecting the VEGF ligand, such as bevacizumab, and those that inhibit the VEGF receptor, such as PTK/ZK. Epidermal growth factor receptor (EGFR) therapies are divided into monoclonal antibodies that affect EGFR, such as cetuximab, and EGFR tyrosine kinase inhibitors, such as gefitinib. Both VEGF and EGFR areas of treatment have shown promising efficacy in first-line, combination therapy settings. Future targeted therapeutic strategies include gene profiling, combinations of capecitabine and oxaliplatin, with bevacizumab and/or cetuximab therapies. PMID- 16360006 TI - Adjuvant therapy of stage II and III colon cancer. AB - Up to 50% of patients who undergo curative resection for colon cancer relapse and ultimately die of metastatic disease. Adjuvant chemotherapy given after surgery reduces the risk of disease recurrence in patients with stage II/III disease, and considerable progress of adjuvant chemotherapy in these patients has been reported. Clinical studies evaluating chemotherapy regimens continue, as do trials that use specific, molecularly targeted agents that have shown promising results in patients with advanced colon cancer. Ongoing important issues to be addressed relate to the optimal schedule and/or choice of chemotherapeutic regimens, and the efficacy of the combination of molecular agents with conventional chemotherapy. PMID- 16360007 TI - Metastatic colorectal cancer: first- and second-line treatment in 2005. AB - First-, second-, and third-line therapies for the treatment of metastatic colorectal cancer may influence choices for subsequent therapy. First-line treatment for metastatic colorectal cancer depends largely on combination chemotherapy regimens that have been proven to prolong survival, control disease progression, and improve quality of life, without excessive toxicity. Standard therapies include 5-fluorouracil, irinotecan, and oxaliplatin, but other combination therapies are currently under investigation. An upcoming area of study includes agents that target vascular endothelial growth factor and epidermal growth factor receptor. While doublet therapy is more active than 5 fluorouracil alone, triplet therapy is also emerging. Second-line therapy is dependent upon prior treatment, and second-line FOLFIRI or FOLFOX regimens that incorporate 5-fluorouracil, leucovorin, and either irinotecan or oxaliplatin, respectively, are still in question because of residual toxicities. After combination chemotherapy, a non-cross-resistant chemotherapy is the best choice. Second-line targeted therapy has been well-tolerated and active in several trials. Future developments will most likely occur in the areas of pharmacogenetics (eg, toxicity, age, comorbidities, patient choice, strategy of cure, palliation) and pharmacogenomics (eg, resistance/activity, gene expression) to produce individualized therapies for patients. The type of adjuvant treatment, genomic consideration, and genetic predisposition will be determining factors in the ideal protocol for first-line therapy. PMID- 16360008 TI - Barrett's Metaplasia: defining the problem. AB - Concern over how best to manage individuals with Barrett's esophagus (BE) has grown because of the consistent rise in the incidence of esophageal adenocarcinoma. Since the 1970s, the rate of increase in incidence of esophageal adenocarcinoma has been greater than that for any other cancer in the US population. Patients with BE have increased risk for esophageal cancer, but the rate of progression and potential risk factors in progression remain poorly understood. Much remains to be learned about BE and its association with adenocarcinoma before effective surveillance or management strategies can be defined and implemented. In this article, the relationship between BE and gastroesophageal reflux disease, risk for adenocarcinoma, and prospects for molecular diagnosis are discussed. PMID- 16360009 TI - Interventional gastroenterology: esophageal and pancreatic cancers. AB - The development and refinement of endoscopic procedures have greatly improved the diagnosis and management of esophageal and pancreatic cancers. Endoscopic ultrasound (EUS) is a highly accurate technique for TNM staging in esophageal cancer, and allows a tissue diagnosis of lymph nodes via fine-needle aspiration with low risk of complications. Endoscopic mucosal resection is a treatment option in patients with early esophageal cancer who are poor surgical candidates. Similarly, EUS fine-needle aspiration is helpful in establishing a diagnosis in cystic lesions, exocrine tumors, neuroendocrine tumors, and other lesions in the pancreas. Endoscopic retrograde cholangiopancreatography provides diagnostic and therapeutic modalities for various pancreaticobiliary problems. A number of promising EUS-guided therapies for pancreatic cancers are under investigation. PMID- 16360010 TI - Transplantation for liver tumors. AB - Currently, the primary use of liver transplantation in the setting of malignancy is in patients with hepatocellular carcinoma, with generally accepted criteria for transplantation consisting of the presence of one nodule less than 5 cm or two of three nodules each less than 3 cm in the absence of detectable vascular invasion. In some patients and settings, surgical resection before transplantation is an emerging, promising option. There is no clear beneficial role of transplantation in patients with resectable or unresectable cholangiocarcinoma, except in selected patients with unresectable disease that is associated with primary sclerosing cholangitis. While good survival results have been achieved with transplantation in patients with epithelioid hemangioendothelioma of the liver, the long-term survival of some patients without any radical treatment leaves the benefit of transplantation unclear. Transplantation would appear to benefit some patients with unresectable liver metastases from neuroendocrine tumors; those who present with non-neuroendocrine liver metastases are not considered candidates for transplantation. PMID- 16360011 TI - Strategies to treat primary unresectable colorectal liver metastases. AB - The liver is the most common site of metastases in patients with colorectal cancer (CRC), and hepatic metastases are responsible for fatalities in at least two thirds of patients with colorectal malignancy. However, the only available treatment associated with long-term survival in patients with CRC metastases is liver resection. While recent studies have shown that liver resection achieves a 5-year overall survival from 37% to 58%, only 10% to 20% of patients with colorectal liver metastases are eligible for resection. Pharmacologic developments and conceptual advances in chemotherapy, regional treatment, and aggressive surgical strategies have ultimately changed the current treatment of patients with primary unresectable liver metastases caused by CRC. Patients who were treated by only palliative chemotherapy a few years ago presently have a variety of strategies available to render their disease surgically resectable with the potential for long-term survival. These advances are the result of a strong collaboration between medical oncologists and surgeons. The development of new chemotherapy protocols that offer the potential for curative surgery with optimum timing within the natural history of this metastatic disease is a shared therapeutic challenge. PMID- 16360012 TI - Humanization of chicken monoclonal antibody using phage-display system. AB - We describe a simple method for humanizing chicken monoclonal antibody (mAb). Humanization of mAbs by simple CDR-grafting often results in loss of affinity because certain framework residues of the antibody variable regions can participate in antigen-antibody interaction. In this study, humanization of chicken mAbs was achieved by CDR-grafting, followed by framework fine-tuning using a chicken phage-displayed mAb, phAb4-31, as a model antibody. In order to fine-tune the framework, we used the phage-displayed combinatorial library with permutation of important framework residues. After panning the humanized library, the "most humanized" variants were selected and analyzed for antigen-binding activity. All of these clones retained affinity comparable to the parental chicken mAb. These results suggest that chicken mAbs can easily be humanized, and thus humanized chicken mAbs may be practically applied as therapeutic agents. PMID- 16360013 TI - The complement receptor 1, CR1 (CD35), mediates inhibitory signals in human T lymphocytes. AB - The modulation the specific, adaptive immune response by complement, particularly of by complement C3, is mainly attributed to its interaction with complement receptors on B-lymphocytes. The function of complement receptors on T lymphocytes, in contrast, is less well understood, although expression of the complement receptor (CR)1 and CR3 on T-cells has been described years ago. In the present study we investigated the effect of antibodies to CR1 on T-cell lines and peripheral T-cells of healthy donors, respectively. Antibodies to CR1 profoundly inhibited the proliferation of the T-cells; of note is, that exogenously added interleukin 2, though enhancing proliferation, did not overcome the inhibitory effect mediated by anti-CR1. While anti-CR1 had no effect on the activation of the immediate early genes c-jun or c-fos nor on the early increase of gamma interferon- or interleukin 2-specific RNA, the protein synthesis of those cytokines was inhibited. Moreover, synthesis of the proliferating cell nuclear antigen (PCNA) was reduced as was the expression of cyclins, particularly of cyclin A and cyclin D3. Taken together, the data indicate that triggering CR1 inhibits proliferation of T-lymphocytes by a mechanism operating downstream of the initial signalling events. PMID- 16360014 TI - The kappa promoter penta-decamer binding protein CBF-A interacts specifically with nucleophosmin in the nucleus only. AB - CArG box-binding factor-A (CBF-A) interacts with the penta-decamer (pd) element that is a conserved sequence element within mouse Vkappa promoters. The pd element acts in synergy with the octamer element to stimulate kappa transcription, especially in later stages of B cell development. To get further insight in the mechanism for CBF-A action we have characterised its protein protein interactions. We show here that CBF-A interacts specifically with nucleophosmin (NPM). This interaction occurs via the homo-oligomerisation domain of NPM and the N-terminus of CBF-A and was exclusive for the nuclear compartment while the two proteins failed to interact in the cytosol. In contrast, CBF-A formed homocomplexes in this compartment. CBF-A was also shown to localise to the nucleoli, most likely dependent on a functional interaction with NPM. Lastly, the sequence fine specificity of CBF-A complexes in the nucleus and cytoplasm were found to differ and nuclear protein-DNA complexes were shown to contain NPM. Thus, CBF-A participates in several protein-protein interactions that may modulate its subcellular localisation and target gene specificity. PMID- 16360015 TI - Cholera toxin induced gene expression alterations. AB - The cholera toxin (CT) is a well-known inducer of cAMP and cAMP regulates gene expression of many genes. However, little is known as to the alterations in gene expression in response to CT. Here the alterations of the expression of 800 selected genes in response to CT were examined using cDNA microarrays. Gene expression alterations in human lymphocytes and monocytes were found after exposure to CT at varying concentrations for different time periods. Over 200 genes showed varying degrees of alterations of expression in CT-treated cells. The CT-induced changes in gene expression were compared by cDNA microarrays under the same conditions to those in response to forskolin, a specific activator of adenylate cyclase, and MDL-12, an irreversible inhibitor of adenylate cyclase. Thirty-five CT-responsive genes were found responded similarly to forskolin but differently to MDL-12. Fourteen CT-responsive genes were affected similarly by MDL-12 but differently by forskolin. Many of these CT responsive genes were involved in immunity, inflammation and oxidative stress. The CT induced responses correlated with those induced by CT subunits. The down regulation of Th1 markers and upregulation of Th2 markers by CT are consistent with the CT induction of Th2 cells. PMID- 16360017 TI - Cloning and biological characterization of buffalo (Bubalus bubalis) interferon gamma. AB - Interferon-gamma, a major immunomodulatory cytokine, of Indian water buffalo (Bubalus bubalis) was characterized at molecular level. Complementary DNA and essential promoter region were cloned and sequenced, and functional recombinant protein was expressed in bacterial system. The cDNA has 97.8% nucleotide identity with 11-nucleotide and four-amino acid variations, and the essential promoter region has 98.4% identity with five-nucleotide variations and a four-nucleotide deletion in comparison with the corresponding bovine sequences. All the major promoter elements such as NF IL-2 like motif, cyclosporin sensitive binding element and GATA motif are strictly conserved. Recombinant buffalo-IFN-gamma expressed in bacterial system reacted with an anti-bovine-IFN-gamma monoclonal antibody in Western blot and showed antiviral activity against buffalo pox virus in cultured Madin-Darby bovine kidney (MDBK) cells by inhibiting virus induced cytopathic effect. The study shows high level sequence similarity of IFN-gamma among ruminants. In view of the immunomodulatory and antiviral activities of IFN gamma, this molecule will be useful in better understanding of the immune system of water buffaloes. PMID- 16360016 TI - Oct-1 DNA binding activity unresponsive to retinoblastoma protein expression prevents MHC class II induction in a non-small cell lung carcinoma cell line. AB - Numerous human tumor lines fail to induce major histocompatibility (MHC) class II expression following interferon-gamma (IFN-gamma) treatment, a response that is considered to be a normal function for almost all human parenchymal and connective tissue cell-types. The effect of MHC class II non-inducibility on solid tumor growth is controversial, but an extensive body of literature indicates that tumor cell MHC class II expression can lead to an antitumor response or tumor tolerance, depending on a number of variables. Thus, understanding the molecular basis for MHC class II induction failures in solid tumor cells will likely lead to ideas for manipulating the antitumor immune response. To date, a handful of tumor associated molecular anomalies have accounted for all the known failures of MHC class II inducibility. In particular, lack of the retinoblastoma tumor suppressor protein (Rb) has been shown in both human and mouse cells to be strongly associated with failure to induce MHC class II. The basis for this relationship is traceable to, among other things, high level Oct-1 DNA binding activity in Rb-defective cells, which represses the prototypical human MHC class II gene, HLA-DRA. Ordinarily, re-establishment of Rb expression leads to elimination of, or substantially reduced Oct-1 DNA binding activity and to rescue of HLA-DRA inducibility. However, in the case of one non small cell lung carcinoma line (NSCLC), Rb re-expression failed to rescue HLA-DRA inducibility despite successful re-establishment of Rb-function. We now report that this failure is traceable to the failure of Rb to rescue normal Oct-1 function. Furthermore, histone deacetylase inhibitor treatment allows a bypass of the Rb requirement and facilitates the MHC class II induction in this NSCLC line. PMID- 16360018 TI - The isolation and characterisation of primary human laryngeal epithelial cells. AB - We have shown that the larynx has a prominent immunological component that varies between individuals, and which is influenced by lifestyle factors implicated in the pathogenesis of the inflammatory and neoplastic diseases of the larynx. In order to explore the mechanisms of such links between laryngeal mucosal immunity and the development of lifestyle-related disease, reliable in vitro models are essential. In this study, we isolated and characterised primary laryngeal epithelial cells from normal individuals and show they can be cultured and manipulated to express MHC class II molecules in vitro. PMID- 16360019 TI - Amino acids critical for binding of autoantibody to an immunodominant conformational epitope of the pyruvate dehydrogenase complex subunit E2: identification by phage display and site-directed mutagenesis. AB - The E2 subunit of the mitochondrial multienzyme pyruvate dehydrogenase complex (PDC-E2) is the major autoantigen in the liver disease, primary biliary cirrhosis (PBC). An epitope region which has been localized to amino acids 91-227 is believed to include the residue K173 to which is attached the lipoyl cofactor. We investigated structural features of this epitope region by screening random peptide phage-displayed libraries and identified prevalent phagotopes that contained likely contact amino acids in separate regions of the linear sequence, H132M133, and F178, V180. These were confirmed by site-directed alanine mutagenesis singly or in combination of the HM and FV residues in wild-type (wt) PDC-E2, and by immunization of rabbits with phage that expressed peptides MHLNTPP or FVLPWRI. The lipoyl lysine K173 also was mutated. Reactivities of mutants and wild-type (wt) PDC-E2, compared by ELISA using 12 PBC sera, showed decremental reactivity of mutant versus wt PDC-E2 (normalized to 100%): wt PDC-E2 (100%)>>PDC E2(F178A,V180A) (mean+/-S.D., 59+/-17%)>PDC-E2(M133A) (50+/-13%)>PDC-E2(H132A) (36+/-13%)>PDC-E2(H132A,M133A) (28+/-8%)>PDC-E2(H132A,M133A,F178V,M180A) (18+/ 13%). Notably PDC-E2(K173A) retained full reactivity (93+/-21%). Rabbits immunized with phage peptides generated antibodies reactive with entire PDC-E2. Our data convincingly validate phage library technology for defining spatially disparate contact residues for conformational epitopes. Ensuing data could be generally applicable to search for occult extrinsic agents as initiators of autoimmunity. PMID- 16360020 TI - The dual specificity phosphatase transcriptome of the murine thymus. AB - Properly regulated mitogen-activated protein (MAP) kinase activity is critical for normal thymocyte development. MAP kinases are activated by phosphorylation of tyrosine and threonine, and dual specificity phosphatases (DUSPs) can inactivate MAP kinases by dephosphorylating both tyrosine and threonine. However, a role for DUSPs in thymocyte development has not been described. In this study, we have defined the subset of DUSP genes expressed in the murine thymus, and how their expression varies in different thymocyte subsets. Of the murine DUSP genes screened that could potentially dephosphorylate MAP kinases, we found 10 transcribed in the thymus. Seven of these 10 thymic DUSPs are true MAP kinase phosphatases based on the presence of a MAP kinase binding domain and demonstrated phosphatase activity against MAP kinases. Six of the seven thymic MAP kinase phosphatases have been shown to dephosphorylate extracellular regulated kinase (ERK). Quantitative PCR analysis of thymocyte populations isolated from different developmental stages revealed significant changes in DUSP expression as thymocytes progressed through development. Specifically, DUSPs 1, 4, and 5 significantly increase in expression as cells go from small, resting CD4/CD8 double positive cells to the CD4 single positive stage. Additionally, in vitro experiments showed that DUSPs could respond to TCR signaling, as anti-CD3 stimulation of thymocytes transiently increased transcription of six of the 10 thymic DUSP genes within 30 min. Notably, the ERK-specific phosphatase DUSP5 was upregulated 43-fold within 30 min, and returned to baseline within 24 h. Overall, we have identified a subset of DUSPs that could potentially regulate ERK activation in response to TCR signals in thymocytes. PMID- 16360021 TI - Induction of regular cytolytic T cell synapses by bispecific single-chain antibody constructs on MHC class I-negative tumor cells. AB - Certain bispecific single-chain antibody constructs exhibit an extraordinary potency for polyclonal T cell engagement and target cell lysis. Here we studied the structural basis for this potency, using laser scanning confocal microscopy. Cytolytic human T cell synapses could be triggered either by addition of a specific peptide antigen or an Ep-CAM-/CD3-bispecific T cell engager (BiTE). Both kinds of synapses showed a comparable distribution of all protein markers investigated. Two other BiTEs constructed from different Ep-CAM-specific antibodies gave similar results. BiTEs could also induce lytic synapses between human T cells and a MHC class I-negative, Ep-CAM cDNA-transfected cell line resulting in potent target cell lysis. This shows that certain T cell recognition molecules on target cells are dispensable for synapse formation and BiTE activity, and suggests that BiTE-activated polyclonal T cells may ignore major immune evasion mechanisms of tumor cells in vivo, such as loss of MHC class I expression. PMID- 16360022 TI - Nanotechnologists seek biological niches. PMID- 16360023 TI - Big Pharma's Commedia. PMID- 16360024 TI - A humble bacterium sweeps this year's Nobel Prize. PMID- 16360025 TI - Tracking RNA polymerase, one step at a time. AB - Although the structural analysis of transcription has attained atomic resolution, techniques to manipulate single molecules involved in transcription have lagged behind. A recent paper in Nature by Abbondanzieri et al. (2005) closes this gap. This study finds that individual molecules of bacterial RNA polymerase move in single base-pair steps and concludes that RNA polymerase acts as a Brownian ratchet that is driven forward by the binding of incoming nucleoside triphosphates. PMID- 16360026 TI - PIDD: a switch hitter. AB - In response to genotoxic stress, the PIDD protein promotes apoptosis downstream of the tumor-suppressor protein p53. In this issue of Cell, Janssens et al. (2005) demonstrate that, in response to such stress, PIDD forms a nuclear complex that enhances sumoylation of NEMO. This modification is important for the activation of the antiapoptotic transcription factor NF-kappaB. PMID- 16360027 TI - Bmp signaling: turning a half into a whole. AB - In a famous experiment over a century ago, Hans Spemann demonstrated that amphibians have a remarkable ability to compensate for perturbations to the embryo. In this issue of Cell, Reversade and De Robertis (2005) uncover the basis of this phenomenon. They demonstrate that interactions between bone morphogenetic proteins (Bmps) and their inhibitors on both the dorsal and ventral sides of the early Xenopus embryo are involved in creating the body plan. PMID- 16360028 TI - Under pressure: stromal fibroblasts change their ways. AB - In this issue of Cell, Hill et al. (2005) demonstrate in a mouse model of prostate cancer that the tumor cells can initiate and promote expansion of stromal fibroblasts that lack the tumor-suppressor protein p53 through a paracrine mechanism. This results in selection of highly proliferative fibroblasts associated with the carcinoma that further promote tumor progression. PMID- 16360029 TI - When size does not matter: pairing sites during meiosis. PMID- 16360030 TI - The many faces of PPARgamma. AB - In an era marked by the increasing prevalence of obesity, diabetes, and cardiovascular disease, the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) has emerged as a transcriptional regulator of metabolism whose activity can be modulated by direct binding of small molecules. As the master regulator of fat-cell formation, PPARgamma is required for the accumulation of adipose tissue and hence contributes to obesity. Yet PPARgamma ligands are clinically effective antidiabetic drugs, although side effects limit their utility. Can PPARgamma be targeted with greater benefit and with less risk to patients? The answer depends upon the basic biology of PPARgamma, and the possibility of selectively modulating the activity of this nuclear receptor in a tissue- and target-gene-specific manner. PMID- 16360031 TI - Selective evolution of stromal mesenchyme with p53 loss in response to epithelial tumorigenesis. AB - Our understanding of cancer has largely come from the analysis of aberrations within the tumor cell population. Yet it is increasingly clear that the tumor microenvironment can significantly influence tumorigenesis. For example, the mesenchyme can support the growth of tumorigenic epithelium. However, whether fibroblasts are subject to genetic/epigenetic changes as a result of selective pressures conferred by oncogenic stress in the epithelium has not been experimentally assessed. Recent analyses of some human carcinomas have shown tumor-suppressor gene mutations within the stroma, suggesting that the interplay among multiple cell types can select for aberrations nonautonomously during tumor progression. We demonstrate that this indeed occurs in a mouse model of prostate cancer where epithelial cell cycle disruption via cell-specific inhibition of pRb function induces a paracrine p53 response that suppresses fibroblast proliferation in associated stroma. This interaction imposes strong selective pressure yielding a highly proliferative mesenchyme that has undergone p53 loss. PMID- 16360032 TI - Ku70, a component of DNA-dependent protein kinase, is a mammalian receptor for Rickettsia conorii. AB - Rickettsia conorii, a strictly intracellular and category C priority bacterial pathogen (NIAID), invades different mammalian cells. Although some signaling events involved in bacterial entry have been documented, the bacterial and host proteins mediating entry were not known. We report the identification of the Ku70 subunit of DNA-dependent protein kinase (DNA-PK) as a receptor involved in R. conorii internalization. Ku70 is recruited to R. conorii entry sites, and inhibition of Ku70 expression impairs R. conorii internalization. Bacterial invasion is dependent on the presence of cholesterol-enriched microdomains containing Ku70. R. conorii infection stimulates the ubiquitination of Ku70. In addition, the ubiquitin ligase c-Cbl is recruited to R. conorii entry foci, and downregulation of endogenous c-Cbl blocks bacterial invasion and Ku70 ubiquitination. An affinity chromatography approach identified the rickettsial protein rOmpB as a ligand for Ku70. This is the first report of a receptor-ligand interaction involved in the internalization of any rickettsial species. PMID- 16360033 TI - Real-time kinetics of gene activity in individual bacteria. AB - Protein levels have been shown to vary substantially between individual cells in clonal populations. In prokaryotes, the contribution to such fluctuations from the inherent randomness of gene expression has largely been attributed to having just a few transcripts of the corresponding mRNAs. By contrast, eukaryotic studies tend to emphasize chromatin remodeling and burst-like transcription. Here, we study single-cell transcription in Escherichia coli by measuring mRNA levels in individual living cells. The results directly demonstrate transcriptional bursting, similar to that indirectly inferred for eukaryotes. We also measure mRNA partitioning at cell division and correlate mRNA and protein levels in single cells. Partitioning is approximately binomial, and mRNA-protein correlations are weaker earlier in the cell cycle, where cell division has recently randomized the relative concentrations. Our methods further extend protein-based approaches by counting the integer-valued number of transcript with single-molecule resolution. This greatly facilitates kinetic interpretations in terms of the integer-valued random processes that produce the fluctuations. PMID- 16360034 TI - Chromosome sites play dual roles to establish homologous synapsis during meiosis in C. elegans. AB - We have investigated the role of pairing centers (PCs), cis-acting sites required for accurate segregation of homologous chromosomes during meiosis in C. elegans. We find that these sites play two distinct roles that contribute to proper segregation. Chromosomes lacking PCs usually fail to synapse and also lack a synapsis-independent stabilization activity. The presence of a PC on just one copy of a chromosome pair promotes synapsis but does not support synapsis independent pairing stabilization, indicating that these functions are separable. Once initiated, synapsis is highly processive, even between nonhomologous chromosomes of disparate lengths, elucidating how translocations suppress meiotic recombination in C. elegans. These findings suggest a multistep pathway for chromosome synapsis in which PCs impart selectivity and efficiency through a "kinetic proofreading" mechanism. We speculate that concentration of these activities at one region per chromosome may have coevolved with the loss of a point centromere to safeguard karyotype stability. PMID- 16360035 TI - HIM-8 binds to the X chromosome pairing center and mediates chromosome-specific meiotic synapsis. AB - The him-8 gene is essential for proper meiotic segregation of the X chromosomes in C. elegans. Here we show that loss of him-8 function causes profound X chromosome-specific defects in homolog pairing and synapsis. him-8 encodes a C2H2 zinc-finger protein that is expressed during meiosis and concentrates at a site on the X chromosome known as the meiotic pairing center (PC). A role for HIM-8 in PC function is supported by genetic interactions between PC lesions and him-8 mutations. HIM-8 bound chromosome sites associate with the nuclear envelope (NE) throughout meiotic prophase. Surprisingly, a point mutation in him-8 that retains both chromosome binding and NE localization fails to stabilize pairing or promote synapsis. These observations indicate that stabilization of homolog pairing is an active process in which the tethering of chromosome sites to the NE may be necessary but is not sufficient. PMID- 16360036 TI - hnRNP K: an HDM2 target and transcriptional coactivator of p53 in response to DNA damage. AB - In response to DNA damage, mammalian cells trigger the p53-dependent transcriptional induction of factors that regulate DNA repair, cell-cycle progression, or cell survival. Through differential proteomics, we identify heterogeneous nuclear ribonucleoprotein K (hnRNP K) as being rapidly induced by DNA damage in a manner that requires the DNA-damage signaling kinases ATM or ATR. Induction of hnRNP K ensues through the inhibition of its ubiquitin-dependent proteasomal degradation mediated by the ubiquitin E3 ligase HDM2/MDM2. Strikingly, hnRNP K depletion abrogates transcriptional induction of p53 target genes and causes defects in DNA-damage-induced cell-cycle-checkpoint arrests. Furthermore, in response to DNA damage, p53 and hnRNP K are recruited to the promoters of p53-responsive genes in a mutually dependent manner. These findings establish hnRNP K as a new HDM2 target and show that, by serving as a cofactor for p53, hnRNP K plays key roles in coordinating transcriptional responses to DNA damage. PMID- 16360037 TI - PIDD mediates NF-kappaB activation in response to DNA damage. AB - Activation of NF-kappaB following genotoxic stress allows time for DNA-damage repair and ensures cell survival accounting for acquired chemoresistance, an impediment to effective cancer therapy. Despite this clinical relevance, little is known about pathways that enable genotoxic-stress-induced NF-kappaB induction. Previously, we reported a role for the p53-inducible death-domain-containing protein, PIDD, in caspase-2 activation and apoptosis in response to DNA damage. We now demonstrate that PIDD plays a critical role in DNA-damage-induced NF kappaB activation. Upon genotoxic stress, a complex between PIDD, the kinase RIP1, and a component of the NF-kappaB-activating kinase complex, NEMO, is formed. PIDD expression enhances genotoxic-stress-induced NF-kappaB activation through augmented sumoylation and ubiquitination of NEMO. Depletion of PIDD and RIP1, but not caspase-2, abrogates DNA-damage-induced NEMO modification and NF kappaB activation. We propose that PIDD acts as a molecular switch, controlling the balance between life and death upon DNA damage. PMID- 16360038 TI - Structure of the Rb C-terminal domain bound to E2F1-DP1: a mechanism for phosphorylation-induced E2F release. AB - The retinoblastoma (Rb) protein negatively regulates the G1-S transition by binding to the E2F transcription factors, until cyclin-dependent kinases phosphorylate Rb, causing E2F release. The Rb pocket domain is necessary for E2F binding, but the Rb C-terminal domain (RbC) is also required for growth suppression. Here we demonstrate a high-affinity interaction between RbC and E2F DP heterodimers shared by all Rb and E2F family members. The crystal structure of an RbC-E2F1-DP1 complex reveals an intertwined heterodimer in which the marked box domains of both E2F1 and DP1 contact RbC. We also demonstrate that phosphorylation of RbC at serines 788 and 795 destabilizes one set of RbC-E2F-DP interactions directly, while phosphorylation at threonines 821 and 826 induces an intramolecular interaction between RbC and the Rb pocket that destabilizes the remaining interactions indirectly. Our findings explain the requirement of RbC for high-affinity E2F binding and growth suppression and establish a mechanism for the regulation of Rb-E2F association by phosphorylation. PMID- 16360039 TI - Mechanism of lysine 48-linked ubiquitin-chain synthesis by the cullin-RING ubiquitin-ligase complex SCF-Cdc34. AB - Ubiquitin chains linked via lysine 48 (K48) of ubiquitin mediate recognition of ubiquitinated proteins by the proteasome. However, the mechanisms underlying polymerization of this targeting signal on a substrate are unknown. Here we dissect this process using the cyclin-dependent kinase inhibitor Sic1 and its ubiquitination by the cullin-RING ubiquitin ligase SCF(Cdc4) and the ubiquitin conjugating enzyme Cdc34. We show that Sic1 ubiquitination can be separated into two steps: attachment of the first ubiquitin, which is rate limiting, followed by rapid elongation of a K48-linked ubiquitin chain. Mutation of an acidic loop conserved among Cdc34 orthologs has no effect on attachment of the first ubiquitin onto Sic1 but compromises the processivity and linkage specificity of ubiquitin-chain synthesis. We propose that the acidic loop favorably positions K48 of a substrate-linked ubiquitin to attack SCF bound Cdc34 approximately ubiquitin thioester and thereby enables processive synthesis of K48-linked ubiquitin chains by SCF-Cdc34. PMID- 16360040 TI - Short telomeres, even in the presence of telomerase, limit tissue renewal capacity. AB - Autosomal-dominant dyskeratosis congenita is associated with heterozygous mutations in telomerase. To examine the dosage effect of telomerase, we generated a line of mTR+/- mice on the CAST/EiJ background, which has short telomeres. Interbreeding of heterozygotes resulted in progressive telomere shortening, indicating that limiting telomerase compromises telomere maintenance. In later generation heterozygotes, we observed a decrease in tissue renewal capacity in the bone marrow, intestines, and testes that resembled defects seen in dyskeratosis congenita patients. The progressive worsening of disease with decreasing telomere length suggests that short telomeres, not telomerase level, cause stem cell failure. Further, wild-type mice derived from the late-generation heterozygous parents, termed wt*, also had short telomeres and displayed a germ cell defect, indicating that telomere length determines these phenotypes. We propose that short telomeres in mice that have normal telomerase levels can cause an occult form of genetic disease. PMID- 16360041 TI - Regulation of ADMP and BMP2/4/7 at opposite embryonic poles generates a self regulating morphogenetic field. AB - Embryos have the ability to self-regulate and regenerate normal structures after being sectioned in half. How is such a morphogenetic field established? We discovered that quadruple knockdown of ADMP and BMP2/4/7 in Xenopus embryos eliminates self-regulation, causing ubiquitous neural induction throughout the ectoderm. ADMP transcription in the Spemann organizer is activated at low BMP levels. When ventral BMP2/4/7 signals are depleted, Admp expression increases, allowing for self-regulation. ADMP has BMP-like activity and signals via the ALK 2 receptor. It is unable to signal dorsally because of inhibition by Chordin. The ventral BMP antagonists Sizzled and Bambi further refine the pattern. By transplanting dorsal or ventral wild-type grafts into ADMP/BMP2/4/7-depleted hosts, we demonstrate that both poles serve as signaling centers that can induce histotypic differentiation over considerable distances. We conclude that dorsal and ventral BMP signals and their extracellular antagonists expressed under opposing transcriptional regulation provide a molecular mechanism for embryonic self-regulation. PMID- 16360042 TI - Cardiac resynchronization therapy: Part 1--issues before device implantation. AB - Cardiac resynchronization therapy (CRT) has been used extensively over the last years in the therapeutic management of patients with end-stage heart failure. Data from 4,017 patients have been published in eight large, randomized trials on CRT. Improvement in clinical end points (symptoms, exercise capacity, quality of life) and echocardiographic end points (systolic function, left ventricular size, mitral regurgitation) have been reported after CRT, with a reduction in hospitalizations for decompensated heart failure and an improvement in survival. However, individual results vary, and 20% to 30% of patients do not respond to CRT. At present, the selection criteria include severe heart failure (New York Heart Association functional class III or IV), left ventricular ejection fraction <35%, and wide QRS complex (>120 ms). Assessment of inter- and particularly intraventricular dyssynchrony as provided by echocardiography (predominantly tissue Doppler imaging techniques) may allow improved identification of potential responders to CRT. In this review a summary of the clinical and echocardiographic results of the large, randomized trials is provided, followed by an extensive overview on the currently available echocardiographic techniques for assessment of LV dyssynchrony. In addition, the value of LV scar tissue and venous anatomy for the selection of potential candidates for CRT are discussed. PMID- 16360043 TI - Cardiac resynchronization therapy: Part 2--issues during and after device implantation and unresolved questions. AB - Encouraged by the clinical success of cardiac resynchronization therapy (CRT), the implantation rate has increased exponentially, although several limitations and unresolved issues of CRT have been identified. This review concerns issues that are encountered during implantation of CRT devices, including the role of electroanatomical mapping, whether CRT implantation should be accompanied by simultaneous atrioventricular nodal ablation in patients with atrial fibrillation, procedural complications, and when to consider surgical left ventricular lead positioning. Furthermore, (echocardiographic) CRT optimization and assessment of CRT benefits after implantation are highlighted. Also, controversial issues such as the potential value of CRT in patients with mild heart failure or narrow QRS complex are addressed. Finally, open questions concerning when to combine CRT with implantable cardioverter-defibrillator therapy and the cost-effectiveness of CRT are discussed. PMID- 16360044 TI - Significance of QRS complex duration in patients with heart failure. AB - Prolongation of QRS (> or =120 ms) occurs in 14% to 47% of heart failure (HF) patients. Left bundle branch block is far more common than right bundle branch block. Left-sided intraventricular conduction delay is associated with more advanced myocardial disease, worse left ventricular (LV) function, poorer prognosis, and a higher all-cause mortality rate compared with narrow QRS complex. It also predisposes heart failure patients to an increased risk of ventricular tachyarrhythmias, but the incidence of cardiac or sudden death remains unclear because of limited observations. A progressive increase in QRS duration worsens the prognosis. No electrocardiographic measure is specific enough to provide subgroup risk categorization for excluding or selecting HF patients for prophylactic implantable cardioverter-defibrillator (ICD) therapy. In ICD patients with HF, a wide underlying QRS complex more than doubles the cardiac mortality compared with a narrow QRS complex. There is a high incidence of an elevated defibrillation threshold at the time of ICD implantation in patients with QRS > or =200 ms. Mechanical LV dyssynchrony potentially treatable by ventricular resynchronization occurs in about 70% of HF patients with left sided intraventricular conduction delay, a fact that would explain the lack of therapeutic response in about 30% of patients subjected to ventricular resynchronization according to standard criteria relying on QRS duration. The duration of the basal QRS complex does not reliably predict the clinical response to ventricular resynchronization, and QRS narrowing after cardiac resynchronization therapy does not correlate with hemodynamic and clinical improvement. Mechanical LV dyssynchrony is best shown by evolving echocardiographic techniques (predominantly tissue Doppler imaging) currently in the process of standardization. PMID- 16360045 TI - Management of heart failure after cardiac resynchronization therapy: integrating advanced heart failure treatment with optimal device function. AB - Cardiac resynchronization therapy (CRT) is an established adjunctive treatment for patients with systolic heart failure (HF) and ventricular dyssynchrony. The majority of recipients respond to CRT with improvements in quality of life, New York Heart Association functional class, 6-min walk test, and ventricular function. Management of HF after CRT may include up-titration of neurohormonal blockade and an exercise prescription through cardiac rehabilitation to further improve and sustain clinical outcomes. Diagnostic data provided by the CRT device may help to facilitate and optimize treatment. Initial nonresponder rates remain problematic. We suggest a simple step-by-step management and troubleshooting strategy that integrates device function with advanced HF therapy in patients who do not initially respond to CRT. This algorithm represents a new, comprehensive, collaborative approach between the HF and electrophysiology specialists to further improve and sustain outcomes in the field of CRT. PMID- 16360046 TI - Why should we care about CARE-HF? AB - Previous trials of cardiac resynchronization therapy (CRT) have suggested that this therapy can significantly improve functional class and exercise capacity during short-term follow-up. The impact of this therapy on morbidity and mortality has only recently been reported. The Cardiac Resynchronization-Heart Failure (CARE-HF) study has definitively shown that CRT significantly reduces mortality (36%, p < 0.002) in patients with NYHA functional class III and IV heart failure and ventricular dyssynchrony. This study also shows that CRT reverses ventricular remodeling and improves myocardial performance progressively for at least 18 months. In heart failure patients, the CARE-HF and Comparison of Medical Therapy, Pacing, and Defibrillation in Heart Failure (COMPANION) (the earlier major morbidity/mortality trial) studies together show the unequivocal benefit for CRT therapy and CRT therapy with back-up defibrillation to significantly reduce mortality and hospitalization compared with optimal medical therapy. Both studies suggest the benefit of adding the implantable cardiac defibrillator to CRT devices, as over one-third of deaths in the CRT-pacemaker arm of both the COMPANION and CARE-HF studies were sudden. PMID- 16360047 TI - There is plenty of room for cardiac resynchronization therapy devices without back-up defibrillators in the electrical treatment of heart failure. AB - Patients with chronic heart failure might benefit from electrical therapy with a view to resynchronize the heart and improve its mechanical performance by cardiac resynchronization therapy (CRT) or to prevent the risk of sudden death by automatic defibrillation. These two therapies can be applied separately or with a combined device, the biventricular implantable cardioverter-defibrillator (CRT D). There is currently no strong scientific evidence indicating that a CRT-D must be offered to all candidates for CRT. Plain common sense should limit the prescription of these costly devices for patients in need of secondary prevention or for younger patients without major comorbidities. The preferential choice of CRT pacemakers in the remainder of patients is currently a logical one. PMID- 16360048 TI - Septal to posterior wall motion delay fails to predict reverse remodeling or clinical improvement in patients undergoing cardiac resynchronization therapy. AB - OBJECTIVES: The aim of this study was to test the hypothesis that a longer septal to-posterior wall motion delay (SPWMD) would predict greater reverse remodeling and an improved clinical response in heart failure patients randomized to cardiac resynchronization therapy (CRT) in the CONTAK-CD trial. BACKGROUND: The SPWMD predicted clinical benefit with CRT in two previous studies from the same center. METHODS: In this retrospective analysis of the CONTAK-CD trial, SPWMD was measured from the baseline echocardiogram of 79 heart failure patients (ejection fraction 22 +/- 7%, QRS duration 159 +/- 27 ms, 72% ischemic, 84% male) randomized to CRT and compared with six-month changes in echocardiographic and clinical parameters. Patients with a left ventricular end-systolic volume index (LVESVI) reduction of at least 15% were considered responders. RESULTS: The feasibility and reproducibility of performing the SPWMD measurements were poor. Larger values for SPWMD did not correlate with six-month changes in left ventricular end-diastolic volume index (p = 0.26), LVESVI (p = 0.41), or left ventricular ejection fraction (p = 0.36). Responders did not have a significantly different SPWMD than non-responders (p = 0.26). The SPWMD did not correlate with measures of clinical improvement. At a threshold of SPWMD >130 ms, the test characteristics to predict reverse remodeling or a clinical response were inadequate. CONCLUSIONS: The previous findings that SPWMD predicts reverse remodeling or clinical improvement with CRT were not reproducible in patients randomized in the CONTAK-CD trial. PMID- 16360049 TI - Propagation of onset and peak time of myocardial shortening in time of myocardial shortening in ischemic versus nonischemic cardiomyopathy: assessment by magnetic resonance imaging myocardial tagging. AB - OBJECTIVES: We aimed to study the relation between onset and peak time of circumferential shortening and the direction of propagation of these parameters in both ischemic and nonischemic patients. BACKGROUND: Peak time is often used to select patients for cardiac resynchronization therapy, whereas pacing influences only the onset times directly. Furthermore, it is unclear whether there is a consistent direction of propagation delay and whether this depends on the etiology. METHODS: Magnetic resonance imaging myocardial tagging with high temporal resolution (14 ms) was applied to 29 patients (18 nonischemic, 11 ischemic) and 17 healthy control subjects. Time to onset (T(onset)), to first peak (T(peak,first)), and to maximum peak (T(peak,max)) of circumferential shortening were determined. Three-dimensional vectors were calculated to denote the main direction of asynchrony. RESULTS: In both patient groups, T(onset) showed a significant positive relation with both T(peak,first) and T(peak,max); however, T(peak,first) correlated considerably better with T(onset) than did T(peak,max) (p < 0.0001 for nonischemic, and p < 0.01 for ischemic patients). Moreover, the relations between T(peak) and T(onset) were stronger in the nonischemic patients than in the ischemic patients (p < 0.001). In nonischemic patients, the propagation of T(onset) was consistently from septum to lateral wall. In the ischemic patients, however, no consistent direction of propagation was found. For both groups, the longitudinal propagation delays (between apex and base) were negligible compared with the short-axis delays. CONCLUSIONS: The relation between peak time and onset time of shortening is strongest in nonischemic patients and is most consistent when time to first peak is used (instead of time to maximum peak). PMID- 16360050 TI - Magnetic resonance imaging assessment of ventricular dyssynchrony: current and emerging concepts. AB - Despite the numerous documented benefits of cardiac resynchronization therapy (CRT), a significant proportion of patients undergoing CRT do not demonstrate symptomatic or morphologic improvement, triggering the search to improve targeting of this therapy. Many studies now support direct assessment of mechanical dyssynchrony as a method to better identify CRT responders. Among the methods used, echo-Doppler imaging has taken center stage and is covered in other articles in this special issue; however, these methods have several inherent limitations, and other alternatives are also being explored such as magnetic resonance imaging (MRI). This review discusses the concepts and clinical use of MRI methods for quantitative assessment of mechanical dyssynchrony, highlighting newer acquisition and analysis methods and focusing on how the data can be synthesized into robust indexes of dyssynchronous heart failure. PMID- 16360051 TI - Electrical signals applied during the absolute refractory period: an investigational treatment for advanced heart failure in patients with normal QRS duration. AB - Cardiac resynchronization therapy has been shown to be an effective treatment for patients with systolic ventricular dysfunction, prolonged (>120 ms) QRS duration, and New York Heart Association (NYHA) functional class III or IV symptoms despite optimal medical therapy. However, studies show that a majority of heart failure patients have QRS duration <120 ms. We have been investigating the potential utility of cardiac contractility modulating (CCM) signals as a treatment option for such patients. Cardiac contractility modulating signals are non-excitatory signals applied during the absolute refractory period using a pacemaker-like device that connects to the heart with pacemaker leads. Acute studies carried out in animals and humans with heart failure suggest that CCM signals can enhance the strength of left ventricular contraction. Results of initial long-term studies designed mainly to demonstrate feasibility and provide preliminary indication of safety in patients with medically refractory NYHA functional class III heart failure are summarized. The results of these preclinical and clinical studies formed the basis for proceeding with two prospective, randomized clinical studies currently underway to definitively test the safety and efficacy of this treatment. PMID- 16360052 TI - Mechanical dyssynchrony assessed by tissue Doppler imaging is a powerful predictor of mortality in congestive heart failure with normal QRS duration. AB - OBJECTIVES: We sought to test whether the mechanical dyssynchrony assessed by tissue Doppler imaging (TDI) is a predictor of cardiac events in patients with congestive heart failure (CHF) and QRS duration < or =120 ms. BACKGROUND: The prevalence and prognostic value of mechanical dyssynchrony in patients with CHF and normal QRS duration have not been well clarified. METHODS: A total of 106 patients (age 63 +/- 11 years) with CHF and ejection fraction (EF) <35% were followed serially; TDI was performed using four basal and four mid-left ventricular segments to assess the time to peak systolic point from R-wave on electrocardiogram (Ts). The standard deviation of Ts (Ts-SD) and the maximal temporal difference of Ts (Ts-diff) of eight segments were used as an indicator of mechanical dyssynchrony. Clinical events included readmission due to worsening of CHF, cardiac transplantation, and death. RESULTS: After 17 +/- 11 months of follow-up, the clinical event rate was 33% including all-cause mortality of 19%. Prolonged Ts-SD (>37 ms) and Ts-diff (>91 ms) were associated with a significant increase in all clinical events. By multivariate analysis, Ts-diff (>91 ms) was an independent risk factor of clinical events and mortality regardless of age, EF, QRS duration, and use of beta-blocking agents. Mean event-free survival was 16.3 months (95% confidence interval [CI] 11.9 to 20.7) in patients with Ts-diff >91 ms and 31.6 months (95% CI 28.0 to 35.1) in those with Ts-diff < or =91 ms, respectively (p < 0.001). CONCLUSIONS: Myocardial dyssynchrony assessed by TDI is a powerful predictor of clinical events in CHF with normal QRS. PMID- 16360053 TI - Improvements in left ventricular diastolic function after cardiac resynchronization therapy are coupled to response in systolic performance. AB - OBJECTIVES: To determine the short-term effects of cardiac resynchronization therapy (CRT) on measurements of left ventricular (LV) diastolic function in patients with severe heart failure. BACKGROUND: Cardiac resynchronization therapy improves systolic performance; however, the effects on diastolic function by load dependent pulsed-wave Doppler transmitral indices has been variable. METHODS: Fifty patients with severe heart failure were evaluated by two-dimensional Doppler echocardiography immediately prior to and 4 +/- 1 month after CRT. Measurements included LV volumes and ejection fraction (EF), pulsed-wave Doppler (PWD)-derived transmitral filling indices (E- and A-wave velocities, E/A ratio, deceleration time [DT], diastolic filling time [DFT], and isovolumic relaxation time). Tissue Doppler imaging was used for measurements of systolic and diastolic (Em) velocities at four mitral annular sites; mitral E-wave/Em ratio was calculated to estimate LV filling pressure. Color M-mode flow propagation velocities were also obtained. RESULTS: After CRT, LV volumes decreased significantly (p < 0.001) and LVEF increased >5% in 28 of 50 patients (56%) and were accompanied by reduction in PWD mitral E-wave velocity and E/A ratio (both p < 0.01), increased DT and DFT (both p < 0.01), and lower filling pressures (i.e., E-wave/Em septal; p < 0.01). Patients with LVEF response < or =5% after CRT had no significant changes in measurements of diastolic function; LV relaxation (i.e., Em velocities) worsened in this group. CONCLUSIONS: In heart failure patients receiving CRT, improvement in LV diastolic function is coupled to the improvement in LV systolic function. PMID- 16360054 TI - Diastolic asynchrony is more frequent than systolic asynchrony in dilated cardiomyopathy and is less improved by cardiac resynchronization therapy. AB - OBJECTIVES: To compare the incidence of diastolic and systolic asynchrony, assessed by tissue Doppler imaging (TDI), in patients with congestive heart failure (CHF) and severe left ventricular (LV) dysfunction, and to assess TDI changes induced by cardiac resynchronization therapy (CRT). BACKGROUND: Thirty percent of CRT candidates are nonresponders. Besides QRS width, the presence of echographic systolic asynchrony has been used to identify future responders. Little is known about diastolic asynchrony and its change after CRT. METHODS: Tissue Doppler imaging was performed in 116 CHF patients (LV ejection fraction 26 +/- 8%). Systolic and diastolic asynchrony was calculated using TDI recordings of right ventricular and LV walls. RESULTS: The CHF group consisted of 116 patients. Diastolic asynchrony was more frequent than systolic, concerning both intraventricular (58% vs. 47%; p = 0.0004) and interventricular (72 vs. 45%; p < 0.0001) asynchrony. Systolic and diastolic asynchrony were both present in 41% patients, but one-third had isolated diastolic asynchrony. Although diastolic delays increased with QRS duration, 42% patients with narrow QRS presented with diastolic asynchrony. Conversely, 27% patients with large QRS had no diastolic asynchrony. Forty-two patients underwent CRT. Incidence of systolic intraventricular asynchrony decreased from 71% to 33% after CRT (p < 0.0001), but diastolic asynchrony decreased only from 81% to 55% (p < 0.0002). Cardiac resynchronization therapy induced new diastolic asynchrony in eight patients. CONCLUSIONS: Diastolic asynchrony is weakly correlated with QRS duration, is more frequent than systolic asynchrony, and may be observed alone. Diastolic asynchrony is less improved by CRT than systolic. Persistent diastolic asynchrony may explain some cases of lack of improvement after CRT despite good systolic resynchronization. PMID- 16360056 TI - Left ventricular dyssynchrony predicts right ventricular remodeling after cardiac resynchronization therapy. AB - OBJECTIVES: The purpose of this research was to evaluate right ventricular (RV) remodeling after six months of cardiac resynchronization therapy (CRT). BACKGROUND: Cardiac resynchronization therapy is beneficial in patients with end stage heart failure. The effect of CRT on RV size is currently unknown. Accordingly, the effects of CRT on RV size, severity of tricuspid regurgitation, and pulmonary artery pressure were evaluated. METHODS: Fifty-six consecutive patients with end-stage heart failure (52% ischemic cardiomyopathy), left ventricular (LV) ejection fraction (EF) < or =35%, QRS duration >120 ms, and left bundle branch block were included. Clinical parameters, LV volumes, LVEF, LV dyssynchrony, and RV chamber size were assessed at baseline and after six months of CRT; LV dyssynchrony was assessed using tissue Doppler imaging. RESULTS: Clinical parameters improved significantly; LV dyssynchrony was acutely reduced after CRT and remained unchanged at six-month follow-up. Left ventricular EF improved significantly from 19 +/- 6% to 26 +/- 8% (p < 0.001), and LV end diastolic volume decreased from 257 +/- 98 ml to 227 +/- 86 ml (p < 0.001). Right ventricular annulus decreased significantly from 37 +/- 9 mm to 32 +/- 10 mm, RV short-axis from 29 +/- 11 mm to 26 +/- 7 mm, and RV long-axis from 89 +/- 11 mm to 82 +/- 10 mm (all p < 0.001). Left ventricular and RV reverse remodeling were only observed in patients with substantial LV dyssynchrony at baseline. Finally, significant reductions in severity of tricuspid regurgitation and pulmonary artery pressure were observed. CONCLUSIONS: Cardiac resynchronization therapy results in significant reverse LV and RV remodeling after six months of CRT in patients with LV dyssynchrony. Moreover, CRT leads to a reduction of the severity of tricuspid regurgitation and a decrease in pulmonary artery pressure. PMID- 16360055 TI - Impact of upgrade to cardiac resynchronization therapy on ventricular arrhythmia frequency in patients with implantable cardioverter-defibrillators. AB - OBJECTIVES: This study compared cardiac resynchronization therapy's (CRT) impact on ventricular tachyarrhythmia susceptibility in patients who, due to worsening heart failure (HF) symptoms, underwent a replacement of a conventional implantable cardioverter-defibrillator (ICD) with a CRT-ICD. BACKGROUND: Cardiac resynchronization therapy is an effective addition to conventional treatment of HF in many patients with left ventricular systolic dysfunction. However, whether CRT-induced improvements in HF status also reduce susceptibility to life threatening arrhythmias is less certain. METHODS: Clinical and ICD electrogram data were evaluated in 18 consecutive ICD patients who underwent an upgrade to CRT-ICD. Pharmacologic HF therapy was not altered during follow-up. The definition of ventricular tachycardia (VT) and ventricular fibrillation (VF) for each patient was as determined by device programming. Statistical comparisons used paired t tests. RESULTS: Findings were recorded during two time periods: 47 +/- 21 months (range 24 to 70 months) before and 14 +/- 2 months (range 9 to 18 months) after CRT upgrade. At time of upgrade, patient age was 69 +/- 11 years and ejection fraction was 21 +/- 8%. Before CRT the frequency of VT, VF, and appropriate ICD shocks was 0.31 +/- 1.23, 0.047 +/- 0.083, and 0.048 +/- 0.085 episodes/month/patient, respectively. After CRT-ICD, VT and VF arrhythmia burdens and frequency of shocks were respectively 0.13 +/- 0.56, 0.001 +/- 0.004, and 0.003 +/- 0.016 episodes/month/patient (p = 0.59, 0.03, and 0.05 vs. pre-CRT). CONCLUSIONS: Arrhythmia frequency and number of appropriate ICD treatments were reduced after upgrade to CRT-ICD for HF treatment. Thus, apart from hemodynamic benefits, CRT may also ameliorate ventricular tachyarrhythmia susceptibility in HF patients. PMID- 16360057 TI - Electrical conduction disturbance effects on dynamic changes of functional mitral regurgitation. AB - OBJECTIVES: The aim of this study was to investigate the relationship between dynamics of functional mitral regurgitation (MR) and the degree of electrical conduction disturbance, and to evaluate the impact of cardiac resynchronization therapy (CRT) on MR severity and its phasic pattern. BACKGROUND: Mechanisms of phasic changes of functional MR, which may be determined by annulus dilation and tethering of the leaflet, remain unclear. METHODS: Transthoracic two-dimensional echocardiography was performed in 60 patients with functional MR. A biventricular pacemaker was implanted in 19 patients. The mitral annulus area (MAA) and the tenting area (TA) were measured from apical views. The MR volume and fraction were assessed by the quantitative pulsed Doppler method. Instantaneous regurgitation flow rate was measured by proximal flow convergence method. A dynamic change in MR flow rate was evaluated by frame-by-frame analysis throughout systole. RESULTS: A phasic pattern with two peaks at early- and late systole and decrease in mid-systole was noticed in 57 patients. The early systolic peak of MR was larger than the late-systolic peak (128.4 +/- 64.3 ml/s vs. 73.9 +/- 55.1 ml/s, p < 0.001). The ratio of flow rate at these two peaks correlated with QRS duration (r = 0.55, p < 0.001). Early-systolic flow rate reduced after CRT (143.9 +/- 60.8 ml/s to 90.7 +/- 54.1 ml/s, p < 0.05), but late systolic flow rate did not (61.5 +/- 55.0 ml/s to 51.2 +/- 40.9 ml/s, p = NS). A similar pattern was observed for TA, whereas MAA did not change after CRT. CONCLUSIONS: Biphasic pattern was found in functional MR, and the ratio of flow rate at two peaks correlated with QRS duration. The CRT decreased regurgitation flow volume by reducing early-systolic MR but not late-systolic MR, resulting in the change in phasic pattern of functional MR. PMID- 16360058 TI - Resynchronization therapy in pediatric and congenital heart disease patients: an international multicenter study. AB - OBJECTIVES: Our objective was to evaluate the short-term safety and efficacy of cardiac resynchronization therapy (CRT) in children. BACKGROUND: Cardiac resynchronization therapy has been beneficial for adult patients with poor left ventricular function and intraventricular conduction delay. The efficacy of this therapy in the young and in those with congenital heart disease (CHD) has not yet been established. METHODS: This is a multi-center, retrospective evaluation of CRT in 103 patients from 22 institutions. RESULTS: Median age at time of implantation was 12.8 years (3 months to 55.4 years). Median duration of follow up was four months (22 days to 1 year). The diagnosis was CHD in 73 patients (71%), cardiomyopathy in 16 (16%), and congenital complete atrioventricular block in 14 (13%). The QRS duration before pacing was 166.1 +/- 33.3 ms, which decreased after CRT by 37.7 +/- 30.7 ms (p < 0.01). Pre-CRT systemic ventricular ejection fraction (EF) was 26.2 +/- 11.6%. The EF increased by 12.8 +/- 12.7 EF units with a mean EF after CRT of 39.9 +/- 14.8% (p < 0.05). Of 18 patients who underwent CRT while listed for heart transplantation, 3 improved sufficiently to allow removal from the transplant waiting list, 5 underwent transplant, 2 died, and 8 others are currently awaiting transplant. CONCLUSIONS: Cardiac resynchronization therapy appears to offer benefit in pediatric and CHD patients who differ substantially from the adult populations in whom this therapy has been most thoroughly evaluated to date. Further studies looking at the long-term benefit of this therapy in this population are needed. PMID- 16360059 TI - Impact of conventional versus biventricular pacing on hemodynamics and tissue Doppler imaging indexes of resynchronization postoperatively in children with congenital heart disease. AB - OBJECTIVES: We sought to evaluate the effects of biventricular (BDOO) pacing compared with conventional (CDOO) atrioventricular (AV) sequential and atrial (AOO) pacing in children and infants in the early postoperative period after open heart surgery for congenital heart disease (CHD). BACKGROUND: Biventricular pacing using right ventricular (RV) and left ventricular (LV) leads can improve hemodynamics in patients with CHD, but it is unclear whether this occurs in early postoperative children with CHD. METHODS: Nineteen children (age, 5 days to 5.4 years; median, 5.5 months) with a definitive biventricular repair for CHD underwent AOO, CDOO, and BDOO pacing with temporary epicardial leads for 10 min each. The AV delay was 80% of the PR interval for the CDOO and BDOO modes. Lead placement was two right atrial, two RV, and one LV. Blood samples for cardiac index (arterial and venous) and tissue Doppler (TDI) traces were obtained in each pacing mode with a Vivid 7 BT04 digital ultrasound system (GE/VingMed, Horten, Norway) from an apical four-chamber view and analyzed with EchoPac software. RESULTS: The QRS duration was significantly shorter for BDOO compared with CDOO, and the cardiac index was higher with BDOO compared with CDOO. Systemic blood pressure was not different between the three modes of pacing (AOO, CDOO, BDOO). The TDI-derived strain rate showed minimal dyssynchrony in AOO as seen by isovolumic tensing (IVT) and peak systolic contraction (PSC) timing differences between RV and LV. The CDOO worsened dyssynchrony with prolonged DeltaIVT and PSC. The BDOO showed improved synchrony as seen by DeltaIVT and PSC. CONCLUSIONS: The TDI-derived strain rate showed worsened ventricular dyssynchrony with CDOO and improvement with BDOO. Cardiac index and QRS duration were improved by BDOO compared with CDOO. This suggests that short-term pacing with BDOO may benefit children with CHD needing pacing in the postoperative period. PMID- 16360060 TI - Resynchronization in pediatrics who needs it? PMID- 16360061 TI - The incremental benefit of rate-adaptive pacing on exercise performance during cardiac resynchronization therapy. AB - OBJECTIVES: The purpose of this research was to investigate the effect of using rate-adaptive pacing and atrioventricular interval (AVI) adaptation on exercise performance during cardiac resynchronization therapy (CRT). BACKGROUND: The potential incremental benefits of using rate-adaptive pacing and AVI adaptation with CRT during exercise have not been studied. METHODS: We studied 20 patients with heart failure, chronotropic incompetence (<85% age-predicted heart rate [AP HR] and <80% HR reserve), and implanted with CRT. All patients underwent a cardiopulmonary exercise treadmill test using DDD mode with fixed AVI (DDD-OFF), DDD mode with adaptive AVI on (DDD-ON), and DDDR mode with adaptive AVI on (DDDR ON) to measure metabolic equivalents (METs) and peak oxygen consumption (VO2max). RESULTS: During DDD-OFF mode, not all patients reached 85% AP-HR during exercise, and 55% of patients had <70% AP-HR. Compared to patients with >70% AP-HR, patients with <70% AP-HR had significantly lower baseline HR (66 +/- 3 beats/min vs. 80 +/- 5 beats/min, p = 0.015) and percentage HR reserve (27 +/- 5% vs. 48 +/ 6%, p = 0.006). In patients with <70% AP-HR, DDDR-ON mode increased peak exercise HR, exercise time, METs, and VO2max compared with DDD-OFF and DDD-ON modes (p < 0.05), without a significant difference between DDD-OFF and DDD-ON modes. In contrast, there were no significant differences in peak exercise HR, exercise time, METs, and VO2max among the three pacing modes in patients with >70% AP-HR. The percentage HR changes during exercise positively correlated with exercise time (r = 0.67, p < 0.001), METs (r = 0.56, p < 0.001), and VO2max (r = 0.55, p < 0.001). CONCLUSIONS: In heart failure patients with severe chronotropic incompetence as defined by failure to achieve >70% AP-HR, appropriate use of rate adaptive pacing with CRT provides incremental benefit on exercise capacity during exercise. PMID- 16360062 TI - Cardiac resynchronization with sequential biventricular pacing for the treatment of moderate-to-severe heart failure. AB - OBJECTIVES: The InSync III study evaluated sequential cardiac resynchronization therapy (CRT) in patients with moderate-to-severe heart failure and prolonged QRS. BACKGROUND: Simultaneous CRT improves hemodynamic and clinical performance in patients with moderate-to-severe heart failure (HF) and a wide QRS. Recent evidence suggests that sequentially stimulating the ventricles might provide additional benefit. METHODS: This multicenter, prospective, nonrandomized, six month trial enrolled a total of 422 patients to determine the effectiveness of sequential CRT in patients with New York Heart Association (NYHA) functional class III or IV HF and a prolonged QRS. The study evaluated: whether patients receiving sequential CRT for six months experienced improvement in 6-min hall walk (6MHW) distance, NYHA functional class, and quality of life (QoL) over control group patients from the reported Multicenter InSync Randomized Clinical Evaluation (MIRACLE) trial; whether sequential CRT increased stroke volume compared to simultaneous CRT; and whether an increase in stroke volume translated into greater clinical improvements compared to patients receiving simultaneous CRT. RESULTS: InSync III patients experienced greater improvement in 6MHW, NYHA functional class, and QoL at six months compared to control (all p < 0.0001). Optimization of the sequential pacing increased (median 7.3%) stroke volume in 77% of patients. No additional improvement in NYHA functional class or QoL was seen compared to the simultaneous CRT group; however, InSync III patients demonstrated greater exercise capacity. CONCLUSIONS: Sequential CRT provided most patients with a modest increase in stroke volume above that achieved during simultaneous CRT. Patients receiving sequential CRT had improved exercise capacity, but no change in functional status or QoL. PMID- 16360063 TI - The hemodynamic effect of intrinsic conduction during left ventricular pacing as compared to biventricular pacing. AB - OBJECTIVES: We sought to investigate the effect of intrinsic conduction over the right bundle on the maximum rate of left ventricular pressure rise (LVdP/dt(max)) during left ventricular (LV) pacing compared to biventricular (BiV) pacing. BACKGROUND: Simultaneous BiV pacing and LV pacing both improve LV function in patients with heart failure and LV asynchrony. We studied the hemodynamic effect of intrinsic conduction leading to ventricular fusion during LV pacing. METHODS: In 34 patients with New York Heart Association functional class III or IV, sinus rhythm with normal atrioventricular (AV) conduction, left bundle branch block, QRS >130 ms, and optimal medical therapy, LVdP/dt(max) was measured invasively during LV and simultaneous BiV pacing. The AV interval was varied in four steps starting (AV1) with an AV interval 40 ms shorter than the intrinsic PQ time and decreased with 25% for each step. RESULTS: At AV1, LVdP/dt(max) was 996 +/- 194 mm Hg/s for LV pacing and 960 +/- 200 mm Hg/s for BiV pacing (p = 0.0009), with all patients showing ventricular fusion during LV pacing. At AV2, 21 patients had ventricular fusion with a LVdP/dt(max) of 983 +/- 213 mm Hg/s and 957 +/- 202 mm Hg/s for LV and BiV pacing, respectively. In the remaining 13 patients without fusion these values were 919 +/- 164 mm Hg/s and 957 +/- 174 mm Hg/s, respectively. The difference between LV and BiV at AV2 is significantly higher when fusion is present (p = 0.01). CONCLUSIONS: The LVdP/dt(max) is higher in LV than in BiV pacing provided that LV pacing is associated with ventricular fusion caused by intrinsic activation. PMID- 16360064 TI - Cost effectiveness of cardiac resynchronization therapy in the Comparison of Medical Therapy, Pacing, and Defibrillation in Heart Failure (COMPANION) trial. AB - OBJECTIVES: The analysis goal was to estimate incremental cost-effectiveness ratios (ICERs) for the Comparison of Medical Therapy, Pacing, and Defibrillation in Heart Failure (COMPANION) trial patients who received cardiac resynchronization therapy (CRT) via pacemaker (CRT-P) or pacemaker-defibrillator (CRT-D) in combination with optimal pharmacological therapy (OPT) relative to patients with OPT alone. BACKGROUND: In the COMPANION trial, CRT-P and CRT-D reduced the combined risk of all-cause mortality or first hospitalization among patients with advanced heart failure and intraventricular conduction delays, but the cost effectiveness of the therapy remains unknown. METHODS: In this analysis, intent-to-treat trial data were modeled to estimate the cost effectiveness of CRT D and CRT-P relative to OPT over a base-case seven-year treatment episode. Exponential survival curves were derived from trial data and adjusted by quality of-life trial results to yield quality-adjusted life-years (QALYs). For the first two years, follow-up hospitalizations were based on trial data. The model assumed equalized hospitalization rates beyond two years. Initial implantation and follow up hospitalization costs were estimated using Medicare data. RESULTS: Over two years, follow-up hospitalization costs were reduced by 29% for CRT-D and 37% for CRT-P. Extending the cost-effectiveness analysis to a seven-year base-case time period, the ICER for CRT-P was 19,600 dollars per QALY and the ICER for CRT-D was 43,000 dollars per QALY relative to OPT. CONCLUSIONS: For the COMPANION trial patients, the use of CRT-P and CRT-D was associated with a cost-effectiveness ratio below generally accepted benchmarks for therapeutic interventions of 50,000 dollars per QALY to 100,000 dollars per QALY. This suggests that the clinical benefits of CRT-P and CRT-D can be achieved at a reasonable cost. PMID- 16360065 TI - Cost effectiveness of cardiac resynchronization therapy. PMID- 16360066 TI - Cardiac resynchronization devices: the Food and Drug Administration's regulatory considerations. AB - Cardiac resynchronization therapy (CRT) devices have been studied clinically since 1998, and have been on the U.S. market since the Food and Drug Administration (FDA) approval of the first product in 2001. Since that time, the FDA has approved many different models from three different manufacturers, representing the first and second generations of these products. All of these products have undergone the FDA pre-market approval process, which examines the safety and effectiveness of the devices for their intended use. Over the last several years, the FDA has adapted recommendations for CRT clinical trials based on an evolving understanding of what these devices can achieve. This paper will outline the dynamic nature of the FDA's approval process for CRT devices and briefly review the clinical trial designs for the first generation devices. PMID- 16360067 TI - Mode of death in advanced heart failure: the Comparison of Medical, Pacing, and Defibrillation Therapies in Heart Failure (COMPANION) trial. AB - OBJECTIVES: The aim of this study was to evaluate the mode of death in patients with advanced chronic heart failure (HF) and intraventricular conduction delay treated with optimal pharmacologic therapy (OPT) alone or OPT with biventricular pacing to provide cardiac resynchronization therapy (CRT) or CRT + an implantable defibrillator (CRT-D). BACKGROUND: Limited data are available on mode of death in advanced HF. No data have existed on mode of death in these patients who also have an intraventricular conduction delay and are treated with CRT or CRT-D. METHODS: Using prespecified definitions and source materials, seven cardiologists assessed mode of death among the 313 deaths that occurred in the Comparison of Medical, Pacing, and Defibrillation Therapies in Heart Failure (COMPANION) trial. RESULTS: A primary cardiac cause was present in 78% of deaths. Pump failure (44.4%) was the most common mode of death followed by sudden cardiac death (SCD) (26.5%). Compared with OPT, CRT-D significantly reduced the number of cardiac deaths (38%, p = 0.006), whereas CRT alone was associated with a non-significant 14.5% reduction (p = 0.33). Both CRT and CRT-D tended to reduce pump failure deaths (29%, p = 0.11 and 27%, p = 0.14, respectively). The CRT-D significantly reduced SCD (56%, p = 0.02), but CRT alone did not. CONCLUSIONS: Pump failure deaths are the predominant mode of death in patients with advanced HF and are modestly reduced by both CRT and CRT-D. Only CRT-D reduced SCD and thus produced a favorable effect on cardiac mortality. PMID- 16360068 TI - Fluoroscopically guided implantation of modern cardiac resynchronization devices: radiation burden to the patient and associated risks. AB - OBJECTIVES: To establish radiation risks for patients undergoing fluoroscopically guided cardiac resynchronization device implantation. BACKGROUND: Cardiac resynchonization therapy (CRT) may be associated with extended fluoroscopic exposure. METHODS: The fluoroscopy time, dose-area product (DAP), exposure parameters, and percentage contribution of the fluoroscopic projections commonly used were recorded in a series of 14 consecutive patients referred for cardiac resynchronization device implantation and compared to corresponding data obtained from a control group of 20 patients who underwent a conventional rhythm device implantation operation. The DAP to peak skin dose, DAP to effective dose, and DAP to gonadal dose conversion factors were determined for biventricular pacing and conventional rhythm device implantation using a humanoid phantom and thermoluminescence dosimetry. RESULTS: The mean total fluoroscopy time and DAP values were 35.2 min and 4,765 cGy cm2, respectively, for biventricular pacing and 8.2 min and 1,106 cGy cm2, respectively, for conventional rhythm device implantation. Patient skin dose from biventricular pacing procedures requiring extended fluoroscopic exposure may exceed threshold dose for the induction of skin effects only if X-ray source-to-skin distance is kept low. The risk values for fatal cancer and severe hereditary disorders, respectively, associated with a typical CRT procedure were 273 per million and 0.2 per million treated patients. CONCLUSIONS: Radiation risks associated with fluoroscopically guided CRT procedures may be considerable. Present data may be used for the estimation of patient radiation risks from CRT procedures performed in other institutions. PMID- 16360069 TI - Potential proarrhythmic effects of biventricular pacing. AB - Resynchronization therapy involving right ventricular endocardial and left ventricular epicardial pacing improves cardiac output, quality of life, and New York Heart Association functional class in patients with congestive heart failure. Although a great deal of attention has been directed at showing the mechanical benefits and in fine-tuning the biventricular pacing configuration and protocol, little attention has been focused on the consequences of reversing the direction of activation of the left ventricular wall. Recent basic science and clinical studies have shown a proarrhythmic effect of reversing the direction of activation of the left ventricular wall. Reversal of the normal activation sequence prolongs the QT interval and increases the existing transmural dispersion of repolarization, creating the substrate and trigger for re-entrant arrhythmias under long QT conditions. A number of case reports of R-on-T extrasystoles and ventricular tachyarrhythmia induction as a result of biventricular pacing support this observation, and raise concern that biventricular pacing may be proarrhythmic in select cases, particularly when associated with a prolonged QT interval. Our focus in this review is on current understanding of transmural heterogeneity of repolarization that exists across the left ventricular wall, how this dispersion of repolarization is amplified as a consequence of reversal of the normal activation sequence, and how these basic experimental findings may apply to patients receiving cardiac resynchronization therapy. PMID- 16360070 TI - Safety of transvenous cardiac resynchronization system implantation in patients with chronic heart failure: combined results of over 2,000 patients from a multicenter study program. AB - OBJECTIVES: The purpose of this study was to evaluate the safety of implanting a cardiac resynchronization therapy (CRT) system. BACKGROUND: Clinicians and patients require data on the safety of the CRT implant procedure to estimate procedural risk. METHODS: We evaluated outcomes of transvenous CRT system implantation in 2,078 patients from the Multicenter InSync Randomized Clinical Evaluation (MIRACLE) study, the MIRACLE Implantable Cardioverter-Defibrillator (ICD) study, and the InSync III study. We compared the MIRACLE study to the InSync III study and the MIRACLE ICD study randomized phase to its general phase to evaluate the effect of new technologies. RESULTS: The implant attempt succeeded in 1,903 of 2,078 (91.6%) patients. Implant time decreased from 2.7 h in the MIRACLE study to 2.3 h in the InSync III study (p < 0.001), and from 2.8 h in the MIRACLE ICD study randomized phase to 2.4 h in the general phase (p < 0.001). The implant procedure produced 62 perioperative complications in 53 (9.3%) MIRACLE trial patients; 159 in 135 (21.1%) MIRACLE ICD study randomized phase patients and 71 in 62 (13.9%) general phase patients (p < 0.05 vs. randomized); and 41 in 37 (8.8%) InSync III study patients (p = NS vs. the MIRACLE study). We observed 73 postoperative complications in 62 (11.7%) MIRACLE trial patients, 77 in 68 (11.9%) MIRACLE ICD study randomized phase patients and 56 in 45 (11.0%) general phase patients (p = NS), and 37 in 34 (8.6%) InSync III study patients (p = NS). A total of 8% of patients required reoperation to treat lead dislodgement, extracardiac stimulation, or infection during follow-up. CONCLUSIONS: Transvenous CRT system implantation appears safe, well-tolerated, has a high success rate, and improves with operator experience and the addition of new technologies. PMID- 16360071 TI - The ultimate payoff. PMID- 16360072 TI - Follow-up of robotically assisted left ventricular epicardial leads for cardiac resynchronization therapy. PMID- 16360073 TI - Augmentative and alternative communication interventions for persons with developmental disabilities: narrative review of comparative single-subject experimental studies. AB - Augmentative and alternative communication (AAC) as an area of clinical and educational practice involves a myriad of decisions related to the symbols, devices, and strategies from which each client must choose. These decision-making points can be productively informed through evidence from comparative intervention studies. The purpose of this review was to synthesize comparative AAC intervention studies using single-subject experimental designs involving participants with developmental disabilities. Following a systematic search, each qualifying study was reviewed in terms of a priori determined appraisal criteria, and summarized in tabular format. Studies were divided into three groups according to their study of aided approaches, unaided approaches, or a combination of both. Results are discussed in terms of methodological adequacy of the studies and their implications for future research and practice. PMID- 16360075 TI - The diagnosis and management of attention-deficit/hyperactivity disorder in preschool children: the state of our knowledge and practice. PMID- 16360077 TI - Sudden death in the young. AB - BACKGROUND: Sudden death is rare in young people, but it has a disproportionate impact on the community. OBJECTIVES: The purpose of this study was to determine the causes of sudden, natural death in persons aged 5-35 years. METHODS: All autopsies conducted at a forensic medicine facility in the years 1995-2004 (inclusive) were reviewed. This facility serves more than 2.5 million people in the eastern part of Sydney, Australia. Data collected included subject age, height, weight, gender, circumstances of death, and pathologic findings at autopsy. Deaths caused by trauma, accidental causes, drowning, and drug toxicity were excluded from analysis. RESULTS: There were 427 nontraumatic, sudden deaths in the 10-year period (70.7% male). Cardiac causes accounted for 56.4%, noncardiac causes 39.3%, and undetermined cause 4.3%. The most common cardiac cause of sudden death was presumed arrhythmia in those with no or minimal structural heart disease (29.0%). Other cardiac causes were acute myocardial infarction (24.5%), myocarditis (11.6%), hypertrophic cardiomyopathy (5.8%), aortic dissection (5.4%), and dilated cardiomyopathy (5.4%). More than two thirds of deaths caused by acute myocardial infarction occurred in the age group from 30 35 years. Sudden cardiac death occurred during physical activity in 10.8% of cases. Sudden cardiac death was reported in a first-degree relative in 4.5% of decedents. The most common noncardiac causes of sudden death were epilepsy (23.8%), intracerebral hemorrhage (23.8%), asthma (16.1%), and pulmonary embolism (12.5%). CONCLUSION: Presumed cardiac arrhythmia is the most common cause of sudden, natural death in the young. There was no reported history of sudden death among the relatives of most decedents. PMID- 16360078 TI - The changing face of sudden cardiac death in the young. PMID- 16360079 TI - Dispersion of repolarization in cardiac resynchronization therapy. AB - BACKGROUND: Proarrhythmic effects of cardiac resynchronization therapy (CRT) as a result of increased transmural dispersion of repolarization (TDR) induced by left ventricular (LV) epicardial pacing in a subset of vulnerable patients have been reported. The possibility of identifying these patients by ECG repolarization indices has been suggested. OBJECTIVES: The purpose of this study was to test whether repolarization indices on the ECG can be used to measure dispersion of repolarization during pacing. METHODS: CRT devices of 28 heart failure patients were switched among biventricular, LV, and right ventricular (RV) pacing. ECG indices proposed to measure dispersion of repolarization were calculated. The effects of CRT on repolarization were simulated in ECGSIM, a mathematical model of electrocardiogram genesis. TDR was calculated as the difference in repolarization time between the epicardial and endocardial nodes of the heart model. RESULTS: PATIENTS: The interval from the apex to the end of the T wave was shorter during biventricular pacing (102 +/- 18 ms) and LV pacing (106 +/- 21 ms) than during RV pacing (117 +/- 22 ms, P < or =.005). T-wave amplitude and area were low during biventricular pacing (287 +/- 125 microV and 56 +/- 22 microV.s, respectively, P = .0006 vs RV pacing). T-wave complexity was high during biventricular pacing (0.42 +/- 0.26, P = .004 vs RV pacing). Simulations: Repolarization patterns were highly similar to the preceding depolarization patterns. The repolarization patterns of different pacing modes explained the observed magnitudes of the ECG repolarization indices. Average and local TDR were not different between pacing modes. CONCLUSION: In patients treated with CRT, ECG repolarization indices are related to pacing-induced activation sequences rather than transmural dispersion. TDR during biventricular and LV pacing is not larger than TDR during conventional RV endocardial pacing. PMID- 16360080 TI - Cost-effectiveness analysis of genetic testing for familial long QT syndrome in symptomatic index cases. AB - BACKGROUND: Genetic testing for long QT syndrome (LQTS) has been available in a research setting for the past decade, and a commercial test has recently become available. However, the costs and effectiveness of genetic testing have not been estimated. OBJECTIVES: The purpose of this study was to conduct a cost effectiveness analysis of genetic testing in the management of patients who have or are suspected to have familial LQTS. METHODS: We examined the incremental cost effectiveness of genetic testing compared with no genetic testing for symptomatic index cases and how this varied according to changes in assumptions and data inputs. Data were obtained from the published literature and a clinical cohort. RESULTS: We found that genetic testing is more cost-effective than not testing for symptomatic index cases at an estimated cost of 2,500 US dollar per year of life saved. These results were generally robust, although they were sensitive to some data inputs such as the cost of testing and the mortality rate among untreated individuals with LQTS. CONCLUSION: A genetic test for familial LQTS is cost-effective relative to no testing, given our assumptions about the population to be tested and the relevant probabilities and costs. The primary benefit of testing is to more accurately diagnose and treat individuals based on a combination of clinical scores and test results. Future economic analyses of testing for familial LQTS should consider the potential benefits of genetic testing of broader populations, including family members. PMID- 16360081 TI - Adenosine-sensitive atrial tachycardia originating from the proximal coronary sinus. AB - BACKGROUND: Atrial tachycardia (AT) can originate from the proximal coronary sinus (CS). However, detailed electrophysiologic characteristics of the tachycardia are not available. OBJECTIVES: We describe the electrophysiologic characteristics, response to adenosine 5'-triphosphate, and results of radiofrequency ablation of AT with the earliest activation in the proximal CS. METHODS: In 7 of 54 patients (age 57 +/- 18 years) with nonmacroreentrant "focal" AT undergoing electrophysiologic study and radiofrequency ablation, the earliest atrial activation site was located in the proximal CS. RESULTS: The earliest activation site was inside the CS 13 +/- 3 mm from the ostium. The AT could be induced and terminated by atrial extrastimuli or burst pacing. In all patients, the AT was also terminated by a very small dose of adenosine 5'-triphosphate (4.2 +/- 1.1 mg). Rapid ventricular pacing during the tachycardia produced ventriculoatrial dissociation. Radiofrequency ablation directed at the earliest atrial activation site was effective in only three patients (group A). In the remaining four patients (group B), after the radiofrequency energy deliveries, the earliest activation site shifted to an adjacent site with a small increase in the cycle length. Three group B patients underwent successful ablation in the slow pathway region. No recurrence was observed over a follow-up period of 22 +/- 5 months. CONCLUSION: AT with earliest activation in the proximal CS is sensitive to a small dose of adenosine 5'-triphosphate. In some patients, radiofrequency applications in the slow pathway region are effective even if the local activation is not early. PMID- 16360082 TI - Safety and efficacy of radiofrequency energy catheter ablation of atrial fibrillation in patients with pacemakers and implantable cardiac defibrillators. AB - BACKGROUND: Catheter ablation has significantly transformed the clinical management of atrial fibrillation (AF). The safety and efficacy of this procedure are not well understood in patients with pacemakers and defibrillators. OBJECTIVES: The purpose of this study was to study the impact of radiofrequency catheter ablation of AF in patients with pacemakers and implantable cardiac defibrillators. METHODS: We studied 86 patients with pacemakers and defibrillators (group I) and a similar number of age- and gender-matched controls (group II) who underwent AF ablation between 1999 and 2004. Clinical and procedural variables were compared between the two groups. In group I, various generator and lead parameters were compared before and after the procedure. Resurgence of clinical AF after 2 months was considered recurrence. RESULTS: Both groups were similar with regard to age, gender, body mass index, and type of AF. Group I had a higher incidence of diabetes (17% vs 6%, P = .03), coronary artery disease (25% vs 13%, P = .05), less prolonged AF (31 +/- 21 vs 45 +/- 30 months, P <.001), lower left ventricular ejection fraction (49 +/- 13% vs 52 +/- 9%, P = .03), and left ventricular end-diastolic dimensions (4.97 +/- 0.81 vs 4.72 +/- 0.67, P = .03). No changes in the sensing and pacing thresholds, impedance of atrial and ventricular leads, or defibrillator coil impedance after AF ablation were observed in group I. Atrial lead dislodgment was seen in two patients. Transient abnormal but "expected" pulse generator behavior was seen in 25% of patients without permanent malfunction. Stroke (1% vs 1%, P = 1.000), pulmonary vein stenosis (2% vs 1%, P = .77), and AF recurrence rates at 12 months were similar between groups I and II, respectively (19% vs 21%, P = .73). CONCLUSION: AF ablation is safe and efficacious in patients with pacemakers and defibrillators. PMID- 16360083 TI - Relationship of the esophagus and aorta to the left atrium and pulmonary veins: implications for catheter ablation of atrial fibrillation. AB - BACKGROUND: A potential complication during ablation of atrial fibrillation (AF) is damage to adjacent structures such as the esophagus and aorta. Fatal atrio esophageal fistulas have developed after surgery- or catheter-based AF ablation procedures. OBJECTIVES: The purpose of this study was to analyze multidetector computed tomography (MDCT) angiographic images to determine the anatomic relationship of the aorta and esophagus to the left atrium (LA). METHODS: Sixty five subjects underwent CT imaging using a 16-slice MDCT scanner: 24 with paroxysmal AF, 21 with chronic AF, and 20 "control" subjects without a history of AF. Measurements assessed included LA diameters, width of the esophagus and aorta in contact with the posterior LA wall, and distance from the esophagus to the four pulmonary veins (PVs), spine, and LA endocardium. RESULTS: Mean LA diameters were significantly larger in patients with AF vs the control group (P = .003 for anteroposterior diameter; P = .009 for transverse diameter). The anterior aspect of the esophagus was directly apposed to the LA in all cases (contact width 18.9 +/- 4.4 mm). The position of the esophagus varied in the posterior mediastinum but on average was closer to the ostia of the left PVs (P = .0001). The descending aorta was in contact with the LA and/or left PVs in 50 of 65 subjects. The esophagus was closer to the spine in the chronic AF vs control group (P = .007), likely due to increased LA dimension. CONCLUSION: In addition to its ability to assess PV anatomy, preprocedural MDCT imaging can investigate the variable relationship of adjacent structures, such as the esophagus and aorta to the LA and PVs. PMID- 16360084 TI - Impact of atrial fibrillation duration on postcardioversion recurrence. AB - BACKGROUND: "Begetting," a mechanistic tenet of atrial fibrillation (AF), stipulates that the rate of recurrence of AF after cardioversion is proportional to the preceding arrhythmia duration. However, recent reports suggest that, for brief durations, the incidence of early recurrence of AF (ERAF) is inversely proportional to duration. These reports were based on potentially biased data. OBJECTIVES: We performed a prospective study to examine the impact of AF duration on postcardioversion recurrence. METHODS: Forty-four patients underwent placement of an implantable cardioverter-defibrillator (ICD) capable of delivering patient elicited AF cardioversion shocks. Subsequently, in the ambulatory setting, the timing of shocks in relationship to perceived AF onset was randomly assigned within individuals to early (as soon as possible) or delayed (1 day later). RESULTS: During a follow-up averaging 199 days per patient, a total of 61 AF episodes among 17 patients occurred for which a patient-elicited cardioversion shock was delivered. Twenty-three shocks were delivered using early protocol (mean 6.8 hours after AF onset), and 38 shocks were delivered using delayed protocol (mean 34.7 hours after AF onset). The incidence of ERAF was significantly lower using the delayed protocol. CONCLUSION: A strategy of approximately 24-hour delay in cardioversion shock timing decreased the incidence of ERAF, relative to a shock delivered within a few hours of AF onset. This observation has important mechanistic and therapeutic implications. PMID- 16360085 TI - Cardiovert!--or better wait a little? Systematic evidence that the initial hours of atrial fibrillation are a "special time". PMID- 16360086 TI - Defibrillation energy requirements using a left anterior chest cutaneous to subcutaneous shocking vector: implications for a total subcutaneous implantable defibrillator. AB - BACKGROUND: Subcutaneous implantable defibrillators (ICDs) are being developed to facilitate ICD implantation. OBJECTIVE: The purpose of this study was to estimate the human defibrillation energy requirement (DER) using a left chest cutaneous (Q) to subcutaneous (SQ) shocking vector. METHODS: Twenty patients undergoing implantation of an indicated ICD were enrolled (15 males, age = 63 +/- 12 years; ejection fraction = 0.27 +/- 0.14). Defibrillation testing was performed using an investigational system consisting of an external defibrillator and a constructed connector to deliver a shock between a pectoral SQ can and a cardiac apical Q electrode. Two attempts at defibrillation using this configuration were allowed. Stage 1 testing started at 70 J with a step-down/step-up to 50 or 100 J, respectively. Stage 2 testing began at 50 J with a step-down/step-up to 30 or 70 J. RESULTS: During stage 1, a 70-J shock was successful in 7/9 (78%) patients. A second attempt was successful in 7/7 patients using a 50-J shock. In the two remaining patients, a second attempt using a 100-J shock was successful. During stage 2, a 50-J shock was successful in 10/11 (91%) patients. The protocol could not be completed in 2/11 patients. Of the remaining nine patients, a second defibrillation was successful in seven (78%) using a 30-J shock. CONCLUSIONS: The defibrillation energy requirement (DER) of this study vector was 50 J or less in most patients. This low DER supports further investigation of a totally SQ-ICD. However, the DER of 100 J in two patients indicates that further investigation is needed regarding DER variability and safety margins. PMID- 16360087 TI - Heterogeneous pulmonary vein myocardial cell repolarization implications for reentry and triggered activity. AB - BACKGROUND: Myocardial cells in the pulmonary veins (PVs) are thought to play a major role in the initiation and maintenance of atrial arrhythmias, including atrial fibrillation. However, systematic single-cell microelectrode recordings from different regions in intact PV-atrial tissues are lacking. OBJECTIVES: The purpose of this study was to determine the transmembrane action potential properties of myocardial cells in different regions of the PV and the left atrium (LA) and assess their arrhythmogenic potential during perfusion with isoproterenol (ISO) and rapid atrial pacing. METHODS: Glass microelectrode recordings of action potentials were made from the left PV and the LA in Langendorff-perfused young (3-4 month) male rats (Fisher344) (n = 9). RESULTS: Action potential duration (APD) of atrial and PV cells had similar duration at a pacing cycle length (CL) of 200 ms. However, shortening of the pacing CL to 100 ms led to heterogeneous repolarization of PV cells. Mid-PV cells had a significantly higher maximum slope of APD restitution than atrial or other PV sites. Intra-PV conduction block developed at rates when LA and proximal PV cells manifested 1:1 capture. Perfusion of ISO and rapid atrial pacing promoted the emergence of early afterdepolarization (EAD) and triggered beats in two out of nine tissues, causing premature atrial activation. No difference in resting potential or AP amplitude could be detected among the PV and LA cells. CONCLUSIONS: PV myocardial cells develop marked heterogeneity in repolarization, and there is a slight ease of developing EAD and triggered activity in response to rapid pacing and ISO infusion. PMID- 16360088 TI - Pulmonary vein cellular electrophysiology and atrial fibrillation: does basic research help us understand clinical pulmonary-vein arrhythmogenesis? PMID- 16360089 TI - Differential beta-adrenoceptor expression induced by nerve growth factor infusion into the canine right and left stellate ganglia. AB - BACKGROUND: Nerve growth factor (NGF) infusion into the right stellate ganglion (RSG) is antiarrhythmic, while NGF infusion into the left stellate ganglion (LSG) is proarrhythmic in dogs with myocardial infarction (MI) and complete atrioventricular block (CAVB). This functional asymmetry suggests differential neural remodeling. OBJECTIVES: To test the hypothesis that NGF infusion into the RSG and the LSG can lead to differential beta-adrenoceptor (beta-AR) expression in dogs with MI and CAVB. METHODS AND RESULTS: We performed immunostaining to quantify beta(1)-AR and beta(3)-AR immunoreactivity in six dogs with MI and CAVB, nine dogs with MI, CAVB, and NGF infusion to the LSG, six dogs with MI, CAVB, and NGF infusion to the RSG, and six normal dogs. There was significantly increased beta(3)-AR immunoreactivity in dogs with NGF infusion into the LSG and significantly decreased beta(3)-AR immunoreactivity in dogs with NGF infusion into the RSG compared with controls and with the MI and CAVB group. There were no significant differences in beta(1)-AR immunoreactivity among these four groups. To determine protein and mRNA expression of beta-ARs, we created MI and CAVB and infused NGF into the LSG in six additional dogs. The noninfarcted left ventricle free wall was harvested 1 week later. The protein level and receptor density of beta(3)-AR (but not beta(1)- or beta(2)-AR) significantly increased in these six dogs compared with normal controls. CONCLUSIONS: We conclude that NGF infusion into the RSG and the LSG in dogs with MI and CAVB induced differential beta(3)-AR expression in the left ventricular myocardium. PMID- 16360090 TI - Beta3-adrenoceptor: friend or foe? PMID- 16360091 TI - Use of thoracic epidural anesthesia for management of electrical storm: a case report. PMID- 16360092 TI - Antiarrhythmic effects of targeted cardiac neuromodulation: is it time for clinical application? PMID- 16360093 TI - Syndactyly and long QT syndrome (CaV1.2 missense mutation G406R) is associated with hypertrophic cardiomyopathy. PMID- 16360094 TI - Entwining electrical and hypertrophic cardiomyopathies. PMID- 16360095 TI - Shock-induced ventricular oversensing due to seal plug damage: a potential mechanism of inappropriate device therapies in implantable cardioverter defibrillators. PMID- 16360097 TI - Cellular alternans as mechanism of cardiac arrhythmogenesis. PMID- 16360096 TI - Cardiac memory: mechanisms and clinical implications. AB - Cardiac memory (CM) is identified as an altered T wave on electrocardiogram and vectorcardiogram that is seen when sinus rhythm resumes after a period of abnormal myocardial activation. Specifically, the sinus rhythm T wave tracks the QRS vector of the abnormal impulse. CM frequently is induced by ventricular pacing or arrhythmias and historically has been considered of minor relevance to medical practice. Although it has long been known that CM can mimic the T-wave inversions of myocardial ischemia, we learned more recently that CM can alter the actions of antiarrhythmic drugs. Furthermore, it provides a template for investigating the mechanisms whereby ventricular pacing affects myocardial physiology. In this article we review the mechanisms believed responsible for induction of CM and some of its more recently recognized clinical manifestations. We also discuss the controversies regarding atrial memory and its potential clinical implications. PMID- 16360098 TI - A long-RP supraventricular tachycardia: what is the mechanism? PMID- 16360099 TI - Premature ventricular contractions causing pacemaker-mediated tachycardia: a failure of postventricular atrial refractory period after premature ventricular contraction extension? PMID- 16360100 TI - Echocardiographic contrast-guided temporary pacemaker placement. PMID- 16360101 TI - Commotio cordis in an athlete. PMID- 16360102 TI - Eccentric retrograde atrial activation in atrioventricular nodal reentrant tachycardia. PMID- 16360105 TI - Development of an internally quenched fluorogenic substrate for kinetic analysis of thioesterases. PMID- 16360106 TI - The importance of using equimolar DNA for transfection analysis of the 5' flanking promoter regions of genes. PMID- 16360107 TI - Enzyme occupancy measurement of intracellular protein tyrosine phosphatase 1B using photoaffinity probes. AB - Protein tyrosine phosphatase 1B (PTP1B) is believed to be one of the enzymes involved in down-regulating the insulin receptor and is a drug target for the treatment of type II diabetes. To better understand the in vitro and in vivo behavior of PTP1B inhibitors, a cell-based assay to directly measure enzyme occupancy of PTP1B by inhibitors using photoaffinity labeling was developed. Two photoaffinity probes were synthesized containing the photolabile diazirine moiety. These photoprobes were specific for PTP1B and T-cell protein tyrosine phosphatase over CD45, with the most potent photoprobe having an IC(50) value of 0.2nM for PTP1B. Activation of the photoprobes with a 40-W UV lamp in the presence of purified AspTyrLysAspAspAspAspLys (Flag)-PTP1B formed a 1:1 irreversible adduct with the enzyme. The photolabeling was competed by known PTP1B inhibitors, vanadate, and the peptide inhibitor N-benzoyl-l-glutamyl-[4 phosphono(difluoromethyl)]-l-phenylalanyl-[4-phosphono(difluoromethyl)]l phenylalanineamide (BzN-EJJ-amide). In HepG2 (human hepatoma cell line) cells, endogenous PTP1B was labeled by the UV-activated photoprobes in both lysed and intact cells. Enzyme occupancy measurements were conducted with a series of PTP1B inhibitors using the photoprobe affinity assay. Several compounds were shown to bind to endogenous PTP1B in the HepG2 intact cells. PMID- 16360108 TI - Development of electrochemical biosensor based on tyrosinase immobilized in composite biopolymeric film. AB - An electrochemical enzyme electrode for dopa and dopamine was developed via an easy and effective immobilization method. The enzyme tyrosinase was extracted from a plant source Amorphophallus companulatus and immobilized in a novel composite of two biopolymers: agarose and guar gum. This composite matrix containing enzyme forms a self-adhering layer on the active surface of glassy carbon electrode, making it a selective and sensitive phenol sensor. Dopa and dopamine were determined by the direct reduction of biocatalytically liberated quinone species at -0.18V versus Ag/AgCl (3M KCl). The analytical characteristics of this sensor, including linear range, lower detection limit, pH, and storage stability, are described. It has reusability up to 15 cycles and a shelf life of more than 2 months. PMID- 16360109 TI - Impact of variable domain glycosylation on antibody clearance: an LC/MS characterization. AB - Variable (Fv) domain N-glycosylation sites are found in approximately 20% of human immunoglobulin Gs (IgGs) in addition to the conserved N-glycosylation sites in the C(H)2 domains. The carbohydrate structures of the Fv glycans and their impact on in vivo half-life are not well characterized. Oligosaccharide structures in a humanized anti-Abeta IgG1 monoclonal antibody (Mab) with an N glycosylation site in the complementary determining region (CDR2) of the heavy chain variable region were elucidated by LC/MS analysis following sequential exoglycosidase treatments of the endoproteinase Lys-C digest. Results showed that the major N-linked oligosaccharide structures in the Fv region have three characteristics (core-fucosylated biantennary oligosaccharides with one or two N glycolylneuraminic acid [NeuGc] residues, zero or one alpha-linked Gal residue, and zero or one beta-linked GalNAc residue), whereas N-linked oligosaccharides in the Fc region contained typical Fc glycans (core-fucosylated, biantennary oligosaccharides with zero to two Gal residues). To elucidate the contribution of Fv glycans to the half-life of the antibody, a method that allows capture of the Mab and determination of its glycan structures at various time points after administration to mice was developed. Anti-Abeta antibody in mouse serum was immunocaptured by immobilized goat anti-human immunoglobulin Fc(gamma) antibody resin, and the captured material was treated with papain to generate Fab and Fc for LC/MS analysis. Different glycans in the Fc region showed the same clearance rate as demonstrated previously. In contrast to many other non-antibody glycosylated therapeutics, there is no strong correlation between oligosaccharide structures in the Fv region and their clearance rates in vivo. Our data indicated that biantennary oligosaccharides lacking galactosylation had slightly faster clearance rates than other structures in the Fv domain. PMID- 16360110 TI - Application of electron spin resonance spin-trapping technique for evaluation of substrates and inhibitors of nitric oxide synthase. AB - The electron spin resonance (ESR) spin-trapping technique coupled with iron dithiocarbamate complexes is one of the most specific methods for nitric oxide (NO) detection. In this study, we applied this method for the evaluation of the substrate and the inhibitors of NO synthase (NOS). A three-line ESR signal was detected from the mixture of inducible NOS (iNOS), l-arginine (Arg), nicotinamide adenine dinucleotide phosphate (NADPH), tetrahydrobiopterin, dithiothreitol, and Fe(2+)-N-(dithiocarboxy) sarcosine (DTCS-Fe), and the signal intensity increased time-dependently. The signal was not observed by excluding either Arg or NADPH, and it was decreased by the addition of hemoglobin, which is an NO scavenger, and N(G)-monomethyl-l-arginine (l-NMMA), N(G)-nitro-l-arginine (l-NAME), and aminoguanidine (AG), which are NOS inhibitors, depending on the concentration. In comparison with l-NAME and AG, l-NMMA strongly inhibited iNOS activity. By using this method, the K(m) value of Arg and the K(i) value of l-NMMA for iNOS were determined to be 12.6 and 6.1muM, respectively. These values are consistent with the reported values measured by the oxyhemoglobin and citrulline assays. These results suggest that the ESR spin-trapping technique coupled with the iron dithiocarbamate complex can be applied for the evaluation of substrates and inhibitors of NOS, and it would be a powerful tool due to its simplicity and high specificity to NO. PMID- 16360111 TI - Quantitation of adenovirus type 5 empty capsids. AB - Adenovirus empty capsids are immature intermediates that lack DNA and viral core proteins. Highly purified preparations of empty and full capsids were generated by subjecting purified adenovirus preparations to repeated cesium chloride gradient separations. PAGE results revealed that empty capsids contain at least five bands that correspond to proteins absent from the mature virus proteome. Peptide mapping by matrix-assisted laser desorption/ionization time-of-flight MS revealed that three of these bands correspond to varying forms of L1 52/55kDa, a protein involved in the encapsidation of the viral DNA. One band at around 31kDa was found to include precursors to proteins VI and VIII. These precursors correspond to proteins that have not been cleaved by the adenovirus-encoded protease and are not present in the mature full capsids. The precursor to protein VIII (pVIII), a capsid cement protein, is used in this study as a marker in reverse-phased HPLC (RP-HPLC) analyses of adenovirus for the quantitation of empty capsids. A novel calculation method applied to the integration of RP-HPLC chromatograms allowed for the generation of a percentage empty capsid value in a given adenovirus preparation. The percentage empty capsid values generated to date by this method show a high degree of precision and good agreement with a cesium chloride gradient/SDS-PAGE quantitation method of empty capsids. The advantage of this method lies in the accurate, precise, and rapid generation of the percentage of empty capsids in a given purified virus preparation without relying on tedious and time-consuming cesium chloride gradient separations and extractions. PMID- 16360112 TI - [Laparoscopic incisional hernia repair: prospective non randomized trial in 51 cases]. AB - AIM OF THE STUDY: Insertion of a mesh in treatment of incisional hernias reduces the risk of recurrence. A single prospective randomized trial have compared laparoscopic and open approach: there were less postoperative complications and fewer recurrences in the laparoscopic group. Aim of this prospective trial was to control these results. PATIENTS AND METHODS: From January 2000 to May 2005, 51 consecutive incisional hernias were operated on by a laparoscopic approach. Incisional hernia was single in 41 and double in 5. It was median in 41 and lateral in 10. Previous hernia repair was noticed in 33.3%. Main criteria was recurrence. We have considered whether one of the following criteria was associated with the risk of recurrence: sex, obesity, previous repair, pre and preoperative sizes of the hernia, uni or multi orificial aspect of the hernia, median or lateral location, mesh size, ratio mesh surface/hernia surface. Others were postoperative mortality and morbidity, duration of hospitalisation and occurrence of late events. RESULTS: At 2 years all patients were followed. Follow up achieved 3 years in 23 cases and 4 years in 9. Recurrence was observed in 7 (13.7%). None predictive factor was disclosed. No death occurred. Median postoperative pain score at D1, D2 and D3 was respectively 3.1+/-1.9, 2.9+/-2.3 and 2.3+/-2.1. Mean postoperative stay was 4.1+/-1.9 days. Seven postoperative complications occurred, al benign. During follow-up 18 events were noticed and of these 8 were chronic abdominal pain. CONCLUSION: This technique could be employed for every type of incisional hernia but peristomial hernias (not assessed in this study) and every patient. Technical improvements ought to be find to reduce recurrence rate. PMID- 16360113 TI - Towards a practical framework for managing the risks of selecting technology to support independent living. AB - Information and communication technology applications can help increase the independence and quality of life of older people, or people with disabilities who live in their own homes. A risk management framework is proposed to assist in selecting applications that match the needs and wishes of particular individuals. Risk comprises two components: the likelihood of the occurrence of harm and the consequences of that harm. In the home, the social and psychological harms are as important as the physical ones. The importance of the harm (e.g., injury) is conditioned by its consequences (e.g., distress, costly medical treatment). We identify six generic types of harm (including dependency, loneliness, fear and debt) and four generic consequences (including distress and loss of confidence in ability to live independently). The resultant client-centred framework offers a systematic basis for selecting and evaluating technology for independent living. PMID- 16360114 TI - Heterologous expression, purification, and properties of human cytochrome P450 27C1. AB - Cytochrome P450 (P450) 27C1 is one of the "orphan" P450 enzymes without a known biological function. A human P450 27C1 cDNA with a nucleotide sequence modified for Escherichia coli usage was prepared and modified at the N-terminus, based on the expected mitochondrial localization. A derivative with residues 3-60 deleted was expressed at a level of 1350nmol/L E. coli culture and had the characteristic P450 spectra. The identity of the expressed protein was confirmed by mass spectrometry of proteolytic fragments. The purified P450 was in the low-spin iron state, and the spin equilibrium was not perturbed by any of the potential substrates vitamin D(3), 1alpha- or 25-hydroxy vitamin D(3), or cholesterol. P450s 27A1 and 27B1 are known to catalyze the 25-hydroxylation of vitamin D(3) and the 1alpha-hydroxylation of 25-hydroxy vitamin D(3), respectively. In the presence of recombinant human adrenodoxin and adrenodoxin reductase, recombinant P450 27C1 did not catalyze the oxidation of vitamin D(3), 1alpha- or 25-hydroxy vitamin D(3), or cholesterol at detectable rates. P450 27C1 mRNA was determined to be expressed in liver, kidney, pancreas, and several other human tissues. PMID- 16360115 TI - Comparison of basic peptides- and lipid-based strategies for the delivery of splice correcting oligonucleotides. AB - Expression of alternatively spliced mRNA variants at specific stages of development or in specific cells and tissues contributes to the functional diversity of the human genome. Aberrations in alternative splicing were found as a cause or a contributing factor to the development, progression, or maintenance of numerous diseases. The use of antisense oligonucleotides (ON) to modify aberrant expression patterns of alternatively spliced mRNAs is a novel means of potentially controlling such diseases. Oligonucleotides can be designed to repair genetic mutations, to modify genomic sequences in order to compensate for gene deletions, or to modify RNA processing in order to improve the effects of the underlying gene mutation. Steric block ON approach have proven to be effective in experimental model for various diseases. Here, we describe our experience in investigating two strategies for ON delivery: ON conjugation with basic peptides and lipid-based particulate system (lipoplex). Basic peptides or Cell Penetrating Peptides (CPP) such as the TAT-derived peptide appear to circumvent many problems associated with ON and drug delivery. This strategy may represent the next paradigm in our ability to modulate cell function and offers a unique avenue for the treatment of disease. Lipoplexes result from the intimate interaction of ON with cationic lipids leading to ON carrying particles able to be taken up by cells and to release ON in the cytoplasm. We have used as an experimental model the correction of a splicing alteration of the mutated beta-globin intron causing thalassemia. Data on cell penetration and efficacy of correction of specific steric block ON delivered either by basic peptides or lipoplex are described. A comparison of the properties of both delivery systems is made respective to the use of this new class of therapeutic molecules. PMID- 16360116 TI - Oligomerization of the Saccharomyces cerevisiae Na+/H+ antiporter Nha1p: implications for its antiporter activity. AB - The Na(+)/H(+) antiporter (Nha1p) from the budding yeast Saccharomyces cerevisiae plays an important role in intracellular pH and Na(+) homeostasis. Here, we show by co-precipitation of differently tagged Nha1p proteins expressed in the same cell that the yeast Nha1p l forms an oligomer. In vitro cross-linking experiments then revealed that Nha1p-FLAG is present in the membranes as a dimer. Differently tagged Nha1p proteins were also co-precipitated from sec18-1 mutant cells in which ER-to-Golgi traffic is blocked under non-permissive temperatures, suggesting that Nha1p may already dimerize in the ER membrane. When we over expressed a mutant Nha1p with defective antiporter activity in cells that also express the wild-type Nha1p-EGFP fusion protein, we found impaired cell growth in highly saline conditions, even though the wild-type protein was appropriately expressed and localized correctly. Co-immunoprecipitation assays then showed the inactive Nha1p-FLAG mutant interacted with the wild-type Nha1p-EGFP protein. These results support the notion that Nha1p exists in membranes as a dimer and that the interaction of its monomers is important for its antiporter activity. PMID- 16360117 TI - Evidence for retarded extinction of aversive learning in anxious children. AB - The habituation to intense acoustic stimuli and the acquisition of differentially conditioned fear were assessed in 53 clinically anxious and 30 non-anxious control children and young adolescents. Anxious children tended to show larger electrodermal responses during habituation, but did not differ in blink startle latency or magnitude. After acquisition training, non-anxious children rated the CS+ as more fear provoking and arousing than the CS- whereas the ratings of anxious children did not differ. However, anxious children rated the CS+ as more fear provoking after extinction, a difference that was absent in non-anxious children. During extinction training, anxious children displayed larger blink magnitude facilitation during CS+ and a trend towards larger electrodermal responses, a tendency not seen in non-anxious children. These data suggest that extinction of fear learning is retarded in anxious children. PMID- 16360118 TI - Prediction of drug and alcohol abuse in hospitalized adolescents: comparisons by gender and substance type. AB - The authors examined psychosocial correlates of drug and alcohol abuse in 462 hospitalized adolescents, and the extent to which these associations may be affected by gender or by substance type. Participants completed a battery of psychometrically-sound, self-report measures of psychological functioning, environmental stress, drug abuse, and alcohol abuse. Four multiple regression analyses were conducted to determine the joint and independent predictors of drug abuse and alcohol abuse, for males and for females. Multiple regression analysis revealed that seven variables--age, depression, impulsivity, low self-esteem, delinquent predisposition, low peer insecurity, and history of child abuse- jointly predicted both drug and alcohol abuse, for both males and females. However, several differences were found with respect to which variables made independent contributions to the predictive models--with only delinquent predisposition making a significant independent contribution for all four conditions. We found distinct patterns of psychosocial predictor variables for drug and alcohol abuse, as well as distinct patterns for males and females. These results may reflect differing risk factors for drug abuse and alcohol abuse in adolescent psychiatric patients--and differing risk factors for males and females. Such differences have potential implications for prevention and treatment. PMID- 16360119 TI - Rosiglitazone protects human neuroblastoma SH-SY5Y cells against acetaldehyde induced cytotoxicity. AB - Acetaldehyde, an inhibitor of mitochondrial function, has been widely used as a neurotoxin because it elicits a severe Parkinson's disease-like syndrome with elevation of the intracellular reactive oxygen species level and apoptosis. Rosiglitazone, a peroxisome proliferator-activated receptor-gamma agonist, has been known to show various non-hypoglycemic effects, including anti-inflammatory, anti-atherogenic, and anti-apoptotic. In this study, we investigated the protective effects of rosiglitazone on acetaldehyde-induced apoptosis in human neuroblastoma SH-SY5Y cells and attempted to examine its mechanism. Acetaldehyde induced apoptosis was moderately reversed by rosiglitazone treatment. Our results suggest that the protective effects of rosiglitazone on acetaldehyde-induced apoptosis may be ascribed to ability to induce the expression of anti-oxidant enzymes and to regulate Bcl-2 and Bax expression. These data indicate that rosiglitazone may provide a useful therapeutic strategy for the prevention of progressive neurodegenerative disease such as Parkinson's disease. PMID- 16360120 TI - Interstitial chromatin alteration causes persistent p53 activation involved in the radiation-induced senescence-like growth arrest. AB - Various stresses including ionizing radiation give normal human fibroblasts a phenotype of senescence-like growth arrest (SLGA), manifested by p53-dependent irreversible G1 arrest. To determine the mechanism of persistent activation of p53, we examined phosphorylated Ataxia telangiectasia mutated (ATM) and phosphorylated histone H2AX foci formation after X-irradiation. Although the multiple tiny foci, detected soon after (<30 min) irradiation, gradually disappeared, some of these foci changed to large foci and persisted for 5 days. Large foci containing phosphorylated ATM and gamma-H2AX co-localized and foci with p53 phosphorylated at serine 15 also showed the same distribution. Interestingly, the signals obtained by telomere fluorescence in situ hybridization (FISH) assay did not co-localize with 90% of the large foci. Our results indicate that chromatin alteration in interstitial chromosomal regions is the most likely cause of continuous activation of p53, which results in the induction of SLGA by ionizing radiation. PMID- 16360121 TI - Evidence for the importance of basal ganglia output nuclei in semantic event sequencing: an fMRI study. AB - Semantic event sequencing is the ability to plan ahead and order meaningful events chronologically. To investigate the neural systems supporting this ability, an fMRI picture sequencing task was developed. Participants sequenced a series of four pictures presented in random order based on the temporal relationship among them. A control object discrimination task was designed to be comparable to the sequencing task regarding semantic, visuospatial, and motor processing requirements but without sequencing demands. fMRI revealed significant activation in the dorsolateral prefrontal cortex and globus pallidus internal part in the picture sequencing task compared with the control task. The findings suggest that circuits involving the frontal lobe and basal ganglia output nuclei are important for picture sequencing and more generally for the sequential ordering of events. This is consistent with the idea that the basal ganglia output nuclei are critical not only for motor but also for high-level cognitive function, including behaviors involving meaningful information. We suggest that the interaction between the frontal lobes and basal ganglia output nuclei in semantic event sequencing can be generalized to include the sequential ordering of behaviors in which the selective updating of neural representations is the key computation. PMID- 16360122 TI - Single exposure to social defeat increases corticotropin-releasing factor and glucocorticoid receptor mRNA expression in rat hippocampus. AB - Stressful life events are able to induce long-term modifications in physiological and neuroendocrine parameters that are related to the onset of several psychiatric disorders. To gain information on molecular modifications involved in long-term changes triggered by stress, we evaluated gene expression in the hippocampus of rats exposed to a single social defeat session. In the social defeat model, the experimental animal is defeated by a dominant male. The defeat induced an increase in body temperature, in distress vocalisations, in serum corticosterone levels and in anxiety-related behaviour measured with an open field test applied 6 h after the exposure to the dominant rat. In the open field test, anxiety-related behaviours were not detectable anymore 30 h after the exposure to the dominant rat and mRNA levels were evaluated at this time-point. The mRNA levels of genes modulated by stress (corticotropin-releasing factor; corticotropin-releasing factor receptor 1; corticotropin-releasing factor binding protein; mineralocorticoid and glucocorticoid receptors; Ca2+/calmodulin dependent protein kinase-like kinase; Krox20; Bcl-2) and control genes (glyceraldehyde-3-phosphate dehydrogenase; beta-actin and cyclophilin A) were measured with real-time reverse transcription polymerase chain reaction. Corticotropin-releasing factor and glucocorticoid receptor mRNA levels were significantly modulated by the stress procedure, both genes showing an increase in rats exposed to a social defeat. No expression level differences were detected for the other genes. In conclusion, we report that 30 h after an acute social stress, a modification in mRNA levels can be detected in rat hippocampus, thus suggesting potential candidate genes involved in mediating long-term responses. PMID- 16360123 TI - Developmental impairment of compound action potential in the optic nerve of myelin mutant taiep rats. AB - The taiep rat is a myelin mutant with an initial hypomyelination, followed by a progressive demyelination of the CNS. The neurological correlates start with tremor, followed by ataxia, immobility episodes, epilepsy and paralysis. The optic nerve, an easily-isolable central tract fully myelinated by oligodendrocytes, is a suitable preparation to evaluate the developmental impairment of central myelin. We examined the ontogenic development of optic nerve compound action potentials (CAP) throughout the first 6 months of life of control and taiep rats. Control optic nerves (ON) develop CAPs characterized by three waves. Along the first month, the CAPs of taiep rats showed a delayed maturation, with lower amplitudes and longer latencies than controls; at P30, the conduction velocity has only a third of the normal value. Later, as demyelination proceeds, the conduction velocity of taiep ONs begins to decrease and CAPs undergo a gradual temporal dispersion. CAPs of control and taiep showed differences in their pharmacological sensitivity to TEA and 4-AP, two voltage dependent K+ channel-blockers. As compared with TEA, 4-AP induced a significant increase of the amplitudes and a remarkable broadening of CAPs. After P20, unlike controls, the greater sensitivity to 4-AP exhibited by taiep ONs correlates with the detachment and retraction of paranodal loops suggesting that potassium conductances could regulate the excitability as demyelination of CNS axons progresses. It is concluded that the taiep rat, a long-lived mutant, provides a useful model to study the consequences of partial demyelination and the mechanisms by which glial cells regulate the molecular organization and excitability of axonal membranes during development and disease. PMID- 16360124 TI - Desensitization of 5-HT2A receptor function by chronic administration of selective serotonin reuptake inhibitors. AB - We have previously shown that chronic treatment with selective serotonin reuptake inhibitors (SSRIs), fluvoxamine and paroxetine, attenuated m chlorophenylpiperazine (mCPP)-induced hypolocomotion in rats. The effect of these SSRIs on the response to mCPP is thought to be caused by the desensitization of 5 HT2C receptor function. In the present study, we investigated whether chronic administration of SSRI could reduce another pharmacological response to mCPP in rats, i.e., the induction of the secretion of corticosterone. The mCPP-induced increase in the serum concentration of corticosterone was not blocked by the 5 HT2C antagonist SB242084, but was blocked by the 5-HT2A antagonist ketanserin. Chronic treatment with fluvoxamine and paroxetine attenuated the response to mCPP, while these SSRIs had no effects in control rats. These results suggest that the desensitization of 5-HT2A receptor function occurs in the same way as that of 5-HT2C receptor function through chronic treatment with either fluvoxamine or paroxetine as a consequence of prolonged exposure to elevated levels of serotonin. The hypersensitivity of 5-HT2A receptors is observed in depressed patients, and chronic treatment with many antidepressants such as tricyclic antidepressants have been reported to reduce 5-HT2A receptor density and/or efficacy. The desensitization of 5-HT2A receptor function might contribute to the therapeutic mechanism of action of these SSRIs, as seen with other classes of antidepressants. PMID- 16360125 TI - Differential effects of SOCS2 on neuronal differentiation and morphology. AB - Neuronal differentiation of neural progenitor cells is regulated by a variety of growth and transcription factors, that not only regulate cell fate of the progenitor cells but that can also regulate neuronal morphology. Suppressor of cytokine signaling-2 (SOCS2) is an intracellular regulator of Growth Hormone (GH) signaling that is expressed in neural stem cells and neurons during development and is required to overcome the inhibitory effects of GH on neuronal differentiation. SOCS2 also promotes neurite outgrowth, however, whether the mechanism by which SOCS2 regulates neuronal differentiation and neurite outgrowth is the same is not clear. Furthermore, whether the over-expression of SOCS2 has physiological in addition to morphological effects is unknown. To address these questions, we differentiated adult neural progenitor cells derived from wildtype C57BL/6 or SOCS2 over-expressing transgenic mice (SOCS2Tg) in the presence or absence of GH and determined effects on neuronal differentiation and morphology. Compared to wildtype cells, differentiation of SOCS2Tg neurospheres resulted in increased neurogenesis, which was not inhibited by GH. The neurons derived from these cells appeared more complex, with increased neurite outgrowth and number. GH did not, however, have any effect on neurite outgrowth of wildtype or SOCS2Tg neurons. Furthermore, basic electrophysiological analysis of wildtype and SOCS2Tg neurons derived from the neurospheres showed that they were both of an immature electrophysiological neuronal phenotype, indicating that although SOCS2 expression can regulate neuronal morphology, it appears to have little effect on neuronal ion channel expression. PMID- 16360126 TI - In the cerebral cortex of female and male mice, amyloid precursor protein (APP) promoter methylation is higher in females and differentially regulated by sex steroids. AB - The over-expression of amyloid precursor protein (APP) gene in certain areas of the brain indicates abnormalities in gene regulation as an important factor for the development of Alzheimer's disease (AD). We have reported recently that APP mRNA expression is lower in female as compared to male and is regulated by sex steroids. As methylation of promoter is crucial for such regulation, we have used isoschizomeric restriction enzymes MspI and HpaII to analyze the pattern of APP promoter methylation in the cerebral cortex of intact, gonadectomized, testosterone- and estradiol-treated adult and old mice of both sexes. Southern blots of DNA digested with HindIII/MspI or HindIII/HpaII from different groups of mice were probed with a 1.27-kb DIG-11-dUTP labeled APP promoter fragment. The results revealed four distinct bands of 315 bp, 596 bp, 911 bp, and 2.6 kb by MspI, a single band of 2.6 kb by HpaII in all groups and an additional 2.9 kb in intact and estradiol treated old male and testosterone-treated adult female mice. The intensity of 2.6-kb band was relatively lower in intact female as compared to male and varied with different treatments in both ages, indicating the modulation of methylation. Thus, these findings showed that APP promoter methylation is higher in female and differentially regulated by sex steroids in the mice cerebral cortex, suggesting a strong correlation between promoter methylation and transcriptional silencing of APP. PMID- 16360127 TI - Separation and identification of neisserial lipooligosaccharide oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection. AB - We determined the optimal conditions for high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) of oligosaccharides (OS) released from neisserial lipooligosaccharides (LOS) by mild acid hydrolysis. We efficiently obtained detailed composition, sequence, and linkage information about high Mr LOS. We found that HPAE-PAD can discriminate isobaric (same Mr) molecules of different structure, for example, nLc4 and Gb4, distinguish alpha from beta chain extensions, and determine the number of phosphoethanolamine (PEA) substituents. HPAE-PAD provided quantitative information that could be used to compare the relative abundances of OS. We used HPAE-PAD to identify all of the known LOS alpha chain antennae. When used with antibody-binding profiles and exoglycosidase digestion results, HPAE-PAD can provide nearly complete structures rapidly. PMID- 16360128 TI - Ion-pairing reversed-phased chromatography/mass spectrometry of heparin. AB - Heparin and heparin-derived components are widely applied anticoagulant drugs used for amongst other applications medical treatment of deep vein thrombosis and pulmonary embolism. Depolymerisation of native heparin results in complex mixtures of sulfated linear oligosaccharides that are usually not well characterised. In order to further characterise such mixtures, two on-line ion pairing reverse-phased chromatography electrospray ionisation (ESI) mass spectrometry methods have been developed. One of the systems allows the determination of more than 200 components in a medium molecular weight heparin preparation, whereas the other system can be used to separate isomeric heparin oligosaccharides after previous separation according to size. This latter system allows semi-preparative isolation of isomeric heparin oligosaccharides. The experimental setup includes on-line cation exchange in order to prevent the ion pairing reagent from entering the mass spectrometer. PMID- 16360129 TI - Relevance of matrix metalloproteinases and their inhibitors after myocardial infarction: a temporal and spatial window. AB - The post-myocardial infarction wound repair process involves temporarily overlapping phases that include inflammation, formation of granulation tissue, scar formation, and overall left ventricle (LV) remodelling. The myocardial extracellular matrix (ECM) plays an important role in maintaining the structural and functional integrity of the heart and is centrally involved in wound repair post-myocardial infarction (MI). The main proteolytic system involved in the degradation of the ECM in the heart is the matrix metalloproteinase (MMPs) system. The present review will focus on the importance of the unique temporal and spatial window of MMPs and their inhibitors (TIMPs) within the different wound healing phases post-MI. It summarizes (1) the MMP/TIMP levels at different time points post-MI, (2) the alterations seen in post-MI healing in genetically modified mice, and (3) the effects and limitations of therapeutic MMP-inhibition post-MI. PMID- 16360130 TI - The enigma of Mayer waves: Facts and models. AB - Mayer waves are oscillations of arterial pressure occurring spontaneously in conscious subjects at a frequency lower than respiration (approximately 0.1 Hz in humans). Mayer waves are tightly coupled with synchronous oscillations of efferent sympathetic nervous activity and are almost invariably enhanced during states of sympathetic activation. For this reason, the amplitude of these oscillations has been proposed as a surrogate measure of sympathetic activity, although in the absence of a clear knowledge of their underlying physiology. Some studies have suggested that Mayer waves result from the activity of an endogenous oscillator located either in the brainstem or in the spinal cord. Other studies, mainly based on the effects of sinoaortic baroreceptor denervation, have challenged this view. Several models of dynamic arterial pressure control have been developed to predict Mayer waves. In these models, it was anticipated that the numerous dynamic components and fixed time delays present in the baroreflex loop would result in the production of a resonant, self-sustained oscillation of arterial pressure. Recent analysis of the various transfer functions of the rat baroreceptor reflex suggests that Mayer waves are transient oscillatory responses to hemodynamic perturbations rather than true feedback oscillations. Within this frame, the amplitude of Mayer waves would be determined both by the strength of the triggering perturbations and the sensitivity of the sympathetic component of the baroreceptor reflex. PMID- 16360131 TI - Arg-Gly-Asp (RGD)-containing peptides increase soluble guanylate cyclase in contractile cells. AB - OBJECTIVES: Alterations in NO/cGMP signaling have been associated with vascular dysfunction. Here, we tested whether peptides containing arginine-glycine aspartic acid (RGD) motifs, commonly found on the binding sites of extracellular matrix to integrins, could increase the expression and function of soluble guanylate cyclase (sGC) in human mesangial cell (HMC), and human aortic smooth muscle (HASMC) cells. METHODS AND RESULTS: Arginine-glycine-aspartic acid-serine (RGDS) promoted an up-regulation in the sGC beta1 subunit steady-state level, both in HMC and HASMC, in a time- and dose-dependent manner. The cellular effects of RGDS-stimulation of sGC expression was an enhanced cellular response to sodium nitroprusside, resulting in elevated cGMP levels and vasodilator-stimulated phosphoprotein (VASP) phosphorylation in both kinds of cells, and an increased NO relaxing effect on cells precontracted with H(2)O(2) or Angiotensin II. Moreover, RGDS was able to restore the sGC levels that had been previously decreased by long term exposure to NO donors. RGDS effects on sGC regulation were due to the specific interaction with alpha(5)beta(1) integrin. To investigate the intracellular mechanisms activated after RGDS cell treatment, pharmacological kinase inhibitors were used. The effect of RGDS on sGC protein content was completely abolished by treating the cells with c-Jun N-terminal kinase (JNK) inhibitors. In addition, c-fos and c-jun were found in the cell nuclei after RGDS treatment, suggesting that the RGDS effect could be mediated by the AP-1 transcription factor. CONCLUSION: Results provide evidence of a mechanism able to increase the sGC protein content linked to increased activity in contractile cells, not only in basal conditions, but also after the down-regulation of the receptor by its own substrate. Elucidation of this novel mechanism provides a rationale for future pharmacotherapy in certain vascular diseases. PMID- 16360132 TI - A direct interaction between TGFbeta activated kinase 1 and the TGFbeta type II receptor: implications for TGFbeta signalling and cardiac hypertrophy. AB - OBJECTIVE: Transforming growth factor beta (TGFbeta)-activated kinase 1 (TAK1) is a MAP kinase kinase kinase involved in numerous signalling pathways and is strongly implicated in cardiac hypertrophy and heart failure. TGFbeta is also associated with hypertension and heart disease, and evidence suggests that TGFbeta1 and TAK1 act together in a cardiac stress signalling pathway. Canonical TGFbeta signalling is mediated through Smad transcription factors, but TGFbeta can also rapidly activate TAK1. The activation of the Smad cascade is well characterised, but little is known about how TAK1 is activated in response to TGFbeta, and no direct link between any MAPK kinase pathway and the TGFbeta receptors has yet been established. Since TAK1 is activated by TGFbeta within 1 min in cardiomyocytes, we hypothesised there might be a direct interaction between TAK1 and one of the TGFbeta receptors. METHODS: We used a combination of in vitro binding assays and co-immunoprecipitation (IP) experiments to investigate whether TAK1 interacted with the type I (ALK1 or ALK5) or type II (TBRII) TGFbeta receptors. Interactions between endogenous proteins were tested using mouse myoblast and rat cardiomyocyte cells. RESULTS: Immunoprecipitation and in vitro binding assays show that TAK1 binds directly to TBRII. Precipitation of endogenous TAK1 protein in rat cardiomyocytes shows that, in addition to a direct association with TBRII, it also interacts indirectly with ALK5. CONCLUSIONS: We describe a novel and specific interaction between TAK1 and TBRII which, for the first time, directly links TAK1 to the TGFbeta signalling cascade and potentially explains how TGFbeta signalling in cardiomyocytes mediates a hypertrophic response. PMID- 16360133 TI - On the impact of NO-globin interactions in the cardiovascular system. AB - During the last years, many reports have provided evidence that the concept of a simple, diffusion-controlled action of NO must be extended by the existence of storage and long-distance transport forms of NO suitable to extend the half-life of the NO radical. In addition, hemoglobin (myoglobin)-dependent formation of NO may account for an additional NO source in the vasculature and in red muscle. On the other hand, there is increasing evidence that the specific subcellular localization of NO synthase isoforms is a critical determinant for their proper biological function. However, it remains obscure how a localized mode of NO action may occur without effective barriers that prevent the diffusion of NO released at a specific site within a cell to other compartments. Members of the globin family of proteins, mainly hemoglobin and myoglobin, have been found to play important roles in all of these processes. PMID- 16360134 TI - T cell priming by tissue-derived dendritic cells: new insights from recent murine studies. AB - Dendritic cells (DCs) act as sentinels in peripheral tissues, continuously scavenging for antigens in their immediate surroundings. Their involvement in T cell responses is generally thought to consist of a linear progression of events, starting with capture of antigen in peripheral tissues such as the skin followed by migration to draining lymphoid organs and MHC-restricted presentation of antigen-derived peptide to induce T cell priming. The role of tissue-derived DCs in the direct priming of immune responses has lately been challenged. It now appears that, at least in some instances, a non-migratory subtype of DCs in the secondary lymphoid tissue presents tissue-derived antigen to T cells. Here, we review recent developments in research on DC function in the priming of immune responses. PMID- 16360135 TI - Quantification of hemokinin-1 peptide production and secretion from mouse B cells. AB - Hemokinin-1 is a recent addition to the family of mammalian tachykinins and is thought to play an important role in B cell and T cell lymphopoiesis. The mRNA coding for this peptide was expressed in some B lymphocyte cell lines including 70Z/3.12, ABE-8.1/2, and RAW8.1 cells, suggesting the possibility that hemokinin 1 may function in an autocrine or paracrine manner in these cells. Therefore, we quantified secretion of this peptide from the 70Z/3.12 cell line expressing hemokinin-1 mRNA. Despite a sensitive radioimmunoassay, we were surprised to find that hemokinin-1 secretion from confluent cells was below the level of detection of this assay. Furthermore, cell lysates routinely demonstrated a low or undetectable immunoreactive peptide. Collectively these studies show a limited production of hemokinin-1 peptide by transformed B cells. PMID- 16360136 TI - Characterization of transferrin receptor-dependent GaC-Tf-FeN transport in human leukemic HL60 cells. AB - BACKGROUND: Understanding the uptake of GaC-Tf-FeN by cells will provide key insights into studies on transferrin-mediated drug delivery. METHODS: The mechanism of GaC-Tf-FeN transporting into and out of HL60 cells has been investigated by comparing transports between GaC-Tf-FeN and apoTf by means of 125I-labeled transferrin. RESULTS: An association constant for GaC-Tf-FeN was 2 times that for apoTf. GaC-Tf-FeN and apoTf of cell surface-bound displayed similar kinetics during the uptake, but the release rates of internalized GaC-Tf FeN and apoTf from cells were different which showed characteristic disparate. The release continued to occur during the incubation of GaC-Tf-FeN in the presence of nonradioactive apoTf. Neither NaN3 nor NH4Cl could completely block internalization of GaC-Tf-FeN, but they prevented the release of GaC-Tf-FeN from the cells. Excess cold unlabeled apoTf could overcome the block in the release due to NH4Cl but not NaN3. The binding and internalization of GaC-Tf-FeN could be competitively inhibited by nonradioactive apoTf. It implies that both bind to the same receptor on the membrane and the localization of GaC-Tf-FeN resembles that of apoTf inside cells. Pretreated cells with pronase abolished the binding of GaC Tf-FeN significantly. CONCLUSION: On the basis of these findings, we proposed the "transferrin receptor" for the mechanism of GaC-Tf-FeN transport by HL60 cells. PMID- 16360137 TI - Pediatric brain natriuretic peptide and N-terminal pro-brain natriuretic peptide reference intervals. AB - BACKGROUND: Brain natriuretic peptide (BNP) and N-terminal pro-brain natriuretic peptide (NT-proBNP) have been shown to be useful biomarkers for the diagnosis of heart failure. Pediatric reference intervals for these analytes have been reported in part. Previous studies lack large numbers in each group, have not covered all age ranges and have not compared results for BNP with NT-proBNP in simultaneously drawn samples. METHODS: We measured BNP in whole blood using the Biosite Triage point-of-care method and plasma NT-proBNP using the Dade RxL Dimension. We assessed between and within-day precision of both methods and after removing outliers employed the Hoffmann approach to calculate pediatric reference intervals over the age range of 0-21 y. We also compared the 2 methods on simultaneously drawn samples. RESULTS: Reference intervals revealed approximately 20-fold higher 97.5th percentiles for neonates than for children >3 y of age. 97.5th percentiles decreased significantly over the first 3 years of life. As shown by others, the CVs for the automated Dade RxL platform were somewhat lower than those for the POCT method. BNP and NT-proBNP correlated well in simultaneously drawn samples (r=0.947). DISCUSSION: Reference intervals for BNP and NT-proBNP are far higher in neonates and infants than in children older than three years of age. The reasons for this are unknown but resemble the elevated CK MBs and troponins also found in neonates, although the 97.5th percentiles for these latter 2 cardiac markers decrease more rapidly to values found in older children by 6 months of age. PMID- 16360138 TI - Five-month-old infants know humans are solid, like inanimate objects. AB - Infants know that humans are exempt from some of the principles that govern the motion of inanimate objects: for instance, humans can be caused to move without being struck. In the current study, we report that infants nevertheless do apply some of the same principles to both humans and objects, where appropriate. Five month-old infants expect humans, like all material objects, to be solid. PMID- 16360139 TI - Agreement and movement: a syntactic analysis of attraction. AB - This paper links experimental psycholinguistics and theoretical syntax in the study of subject-verb agreement. Three experiments of elicited spoken production making use of specific characteristics of Italian and French are presented. They manipulate and examine its impact on the occurrence of 'attraction' errors (i.e. incorrect agreement with a word that is not the subject of the sentence). Experiment 1 (in Italian) shows that subject modifiers do not trigger attraction errors in free inverted VS (Verb Subject) structures, although attraction was found in VS interrogatives in English (Vigliocco, G., & Nicol, J. (1998). Separating hierarchical relations and word order in language production. Is proximity concord syntactic or linear? Cognition, 13-29) In Experiment 2 (in French), we report stronger attraction with preverbal clitic object pronouns than with subject modifiers. Experiment 3 (in French) shows that displaced direct objects in the cleft construction trigger attraction effects, in spite of the fact that the object does not intervene between the subject and the verb in the surface word order (OSV). Moreover, attraction is stronger in structures with subject-verb inversion (...). These observations are shown to be naturally interpretable through the tools of formal syntax, as elaborated within the Principles and Parameters/Minimalist tradition. Three important constructs are discussed: (1) the hierarchical representation of the sentence during syntactic construction, and the role of intermediate positions by which words transit when they move; (2) the role of specific hierarchical (c-command) but also linear (precedence) relations; and (3) the possibility that agreement involves two functionally distinct components. A gradient of computational complexity in agreement is presented which relates empirical evidence to these theoretical constructs. PMID- 16360140 TI - Notch signaling is required for normal prostatic epithelial cell proliferation and differentiation. AB - Notch pathway is crucial for stem/progenitor cell maintenance, growth and differentiation in a variety of tissues. Using a transgenic cell ablation approach, we found in our previous study that cells expressing Notch1 are crucial for prostate early development and re-growth. Here, we further define the role of Notch signaling in regulating prostatic epithelial cell growth and differentiation using biochemical and genetic approaches in ex vivo or in vivo systems. Treatment of developing prostate grown in culture with inhibitors of gamma-secretase/presenilin, which is required for Notch cleavage and activation, caused a robust increase in proliferation of epithelial cells co-expressing cytokeratin 8 and 14, lack of luminal/basal layer segregation and dramatically reduced branching morphogenesis. Using conditional Notch1 gene deletion mouse models, we found that inactivation of Notch1 signaling resulted in profound prostatic alterations, including increased tufting, bridging and enhanced epithelial proliferation. Cells within these lesions co-expressed both luminal and basal cell markers, a feature of prostatic epithelial cells in predifferentiation developmental stages. Microarray analysis revealed that the gene expression in a number of genetic networks was altered following Notch1 gene deletion in prostate. Furthermore, expression of Notch1 and its effector Hey-1 gene in human prostate adenocarcinomas were found significantly down-regulated compared to normal control tissues. Taken together, these data suggest that Notch signaling is critical for normal cell proliferation and differentiation in the prostate, and deregulation of this pathway may facilitate prostatic tumorigenesis. PMID- 16360141 TI - The effect of long-term low-dose lead exposure on thyroid function in adolescents. AB - This study investigated blood lead (Pb-B) levels and Pb-B effects on thyroid functions in long-term low-level-lead-exposed male adolescents who work as auto repairers. Pb-B and ALAD index (logarithm of activated delta-aminolaevulinic acid dehydratase/nonactivated delta-aminolaevulinic acid dehydratase) were measured as indicators of exposure to lead. Thyroid function tests including free thyroxine (FT4), free triiodothyronine (FT3), and thyrotrophin (TSH) were conducted and thyroid ultrasounds were performed in 42 lead-exposed adolescents and 55 healthy control subjects. Mean Pb-B levels and ALAD index were found significantly higher in the study group than in the normal control group (7.3+/-2.92 microg/dl vs. 2.08+/-1.24 microg/dl, P < 0.001 and 0.44+/-0.26 vs. 0.29+/-0.23, P < 0.05, respectively). FT4 levels were found significantly lower in the study group (1.02+/-0.18 mI/mL and 1.12+/-0.14 mIU/mL, P < 0.05). No subject in the control group had an abnormal FT4 level, but FT4 levels were found under normal limits in 11 subjects (26%) in the study group. FT3 and TSH levels in the study and control groups did not differ (P > 0.05). Thyroid volumes in the study and control groups did not exhibit any significant differences (P > 0.05). Pb-B was found to be negatively correlated to FT4 levels (r = -0.20, P = 0.044). This study revealed that long-term low-level lead exposure may lead to reduced FT4 level without significant changes in TSH and T3 levels in adolescents even at low Pb-B levels. PMID- 16360142 TI - Maternal contamination with dichlorodiphenyltrichloroethane and reproductive outcomes in an Australian population. AB - Persistent organochlorine pesticides, such as dichlorodiphenyltrichloroethane (DDT), are lipophilic environmental pollutants which accumulate in the food chain. These chemicals have long half-lives and can be detected in human milk, serum, and some other tissues. These chemicals have recently been under scrutiny for their possible health hazards such as cancer and reproductive outcomes including low birth weight. The aim of our study was to investigate whether mothers with a higher contamination of pesticides were different from mothers with low contamination in relation to their offspring's birth outcomes such as birth weight, small for gestation age, prematurity, head circumference, sex ratio, and previous miscarriage or still birth. We used data collected as part of a cross-sectional study of organochlorines in breast milk. This study did not show any association between low birth weight or small for gestation age and organochlorine contamination when comparing the higher to the low exposure group. Weak, significant correlations (-0.1) were detected between low birth weight and contamination levels of dichlorodiphenyldichloroethylene (DDE) only in female offspring. There was a decrease in the percentage of female offspring for all chemicals in the high-contamination group. The baby's head circumference increased as the mother's contamination increased but the adjusted difference in means was not significant. We did not see any association between either miscarriage or stillbirth in the mother's previous pregnancies or prematurity of the first live (recent) baby when comparing the higher to the low exposure mothers. The organochlorines DDT and DDE were not found to be associated with adverse birth outcomes in contaminated mothers in the range of contamination of our population (<7.5 mg/kg lipid in maternal milk), although there is weak evidence that sex ratio may be affected. PMID- 16360143 TI - Low blood lead concentration in association with infertility in women. AB - This study investigated the relationship between low-level lead exposure and risk of infertility in women. Study participants consisted of 64 cases recruited at a private infertility clinic and 83 control women from the postpartum clinic unit at a medical center in Kaohsiung, Taiwan. With informed consent, venous blood samples were obtained from subjects, during their early follicular phases (the 1st to 5th days of the menstrual cycle), for measuring concentrations of lead and hormones. Information on other potential risk factors associated with infertility, including reproductive history, Chinese herbal Medicine use, dairy product consumption, residential environment, and sociodemographic status, was obtained during an in-person interview. The mean blood lead level (BLL) in infertile women was significantly higher than that in controls (3.55 vs 2.78 microg/dL, P=0.007). Compared to women with BLL < or = 2.5 microg/dL, women with BLL>2.5 microg/dL were associated with a threefold increased risk for infertility (adjusted odds ratio=2.94, 95% confidence interval 1.18-7.34), after controlling for age, body mass index, smoking, Chinese herbal Medicine use, and irregular menstruation. Women's BLL was a significant predictor of the serum estradiol concentration also. These findings suggest an important role of very low BLL in the risk of infertility in women. PMID- 16360144 TI - Activation of connective tissue-type and mucosal-type mast cells in compound 48/80-induced airway response. AB - The pathology of non-immunological airway contraction is not well understood. To define the activation of different phenotypes of mast cells, a rat non immunological asthmatic model was prepared. Airway contraction in rats was measured by an unrestrained whole-body plethysmographic system following a 10-min inhalation challenge with a 5% solution of compound 48/80. Histamine, leukotrein C(4) (LTC(4)) and tumor necrosis factor (TNF)-alpha levels in bronchoalveolar lavage fluid, as well as tissue histamine content were quantified. Mast cells and eosinophils were detected by histology. Both the early and late phase of airway responses were induced by inhalation of compound 48/80. Histamine and TNF-alpha levels increased significantly 30 min after challenge, but no increases were detected at either 8 or 24 h after challenge. A high LTC(4) level was detected in 30 min and 8 h after challenge. Tissue histamine content decreased at 30 min after challenge and returned to the unstimulated level by 8 h. Connective tissue mast cells in rat trachea showed a degranulation response. Along with the increase in numbers of mucosal mast cells, rat mast cell protease II at both mRNA and protein levels in the trachea epithelial layer was also increased significantly at 30 min after challenge. We conclude that compound 48/80 inhalation causes both the early and late phase of airway contraction in rats. Mast cell degranulation is responsible for the early phase of airway response, which subsequently triggers the late phase of airway response. PMID- 16360145 TI - Effects of systemic administration of beta-casomorphin-5 on learning and memory in mice. AB - The effects of systemic administration of bovine beta-casomorphin-5 (Tyr-Pro-Phe Pro-Gly), a mu-opioid receptor agonist derived from milk beta-casein, on spontaneous alternation behavior in the Y-maze (spatial short-term memory) and step-down-type passive avoidance response (non-spatial long-term memory) were investigated in mice. Intraperitoneal (i.p.) administration of beta-casomorphin-5 (0.1-20 mg/kg) did not have a significant effect on either spontaneous alternation behavior or passive avoidance response. However, a low dose (1 mg/kg, i.p.) of beta-casomorphin-5 improved scopolamine (1 mg/kg, s.c.)-induced impairment of spontaneous alternation behavior and passive avoidance response. Pretreatment with intracerebroventricular injections of beta-funaltrexamine (a mu opioid receptor antagonist, 0.1 microg/mouse) and naloxonazine (a mu(1)-opioid antagonist, 5 microg/mouse), which did not improve scopolamine-induced impairment, prevented the ameliorating effect of beta-casomorphin-5 on scopolamine-induced impairment of passive avoidance response. These results indicated that systemic administration of a low dose (1 mg/kg, i.p.) of beta casomorphin-5 improves the disturbance of learning and memory resulting from cholinergic dysfunction through central mediation involving mu(1)-opioid receptors. PMID- 16360147 TI - Lipid products of phosphoinositide 3-kinase abrogate genistein-induced fusion inhibition in myoblasts. AB - Genistein (4',5,7-trihydroxyisoflavone) is a tyrosine kinase inhibitor. Although the agent has shown to inhibit myoblast differentiation, neither intracellular target(s) as a tyrosine kinase inhibitor nor action mechanism of the agent is well known. Here we studied the effect of genistein on the differentiation of myoblasts. Genistein strongly but reversibly blocked both myoblast fusion and synthesis of the muscle-specific proteins. The agent also reversibly reduced the phosphorylation level of focal adhesion kinase (FAK), a cytoplasmic tyrosine kinase, and its interaction with p85, the regulatory subunit of phosphoinositide 3-kinase (PI3-kinase). In addition, genistein indirectly inhibited PI3-kinase activity and blocked calcium influx which is required for myoblast fusion. However, both genistein-induced inhibition of cell fusion and calcium influx were abrogated by the lipid products of PI3-kinase. These results demonstrate that genistein can exert their effect on the signaling pathway from FAK to calcium influx via PI3-kinase in the differentiation of myoblasts. PMID- 16360146 TI - Capsaicin-induced, capsazepine-insensitive relaxation of the guinea-pig ileum. AB - The mechanisms underlying transient receptor potential vanilloid receptor type 1 (TRPV1)-independent relaxation elicited by capsaicin were studied by measuring isometric force and phosphorylation of 20-kDa regulatory light chain subunit of myosin (MLC(20)) in ileum longitudinal smooth muscles of guinea-pigs. In acetylcholine-stimulated tissues, capsaicin (1-100 microM) and resiniferatoxin (10 nM-1 microM) produced a concentration-dependent relaxation. The relaxant response was attenuated by 4-aminopyridine and high-KCl solution, but not by capsazepine, tetraethylammonium, Ba(2+), glibenclamide, charybdotoxin plus apamin nor antagonists of cannabinoid receptor type 1 and calcitonin-gene related peptide. A RhoA kinase inhibitor reduced the relaxant effect of capsaicin at 30 microM. Capsaicin and resiniferatoxin reduced acetylcholine- and caffeine-induced transient contractions in a Ca(2+)-free, EGTA solution. Capsaicin at 30 microM for 20 min did not alter basal levels of MLC(20) phosphorylation, but abolished an increase by acetylcholine in MLC(20) phosphorylation. It is suggested that the relaxant effect of capsaicin at concentrations used is not mediated by TRPV1, but by 4-aminopyridine-sensitive K(+) channels, and that capsaicin inhibits contractile mechanisms involving Ca(2+) release from intracellular storage sites. The relaxation could be explained by a decrease in phosphorylation of MLC(20). PMID- 16360148 TI - Role of periaqueductal grey prostaglandin receptors in formalin-induced hyperalgesia. AB - In this study we have investigated the role of periaqueductal grey prostaglandin receptors in formalin-induced hyperalgesia in mice. Glutamate and GABA release changes have been monitored by in vivo microdialysis. Intra-periaqueductal grey microinjections of misoprostol, a non-selective prostaglandin receptor agonist, increased nociceptive responses in the formalin test only during the late phase. Prostanoid EP(1) (L-335677), EP(2) (AH 6809), EP(3) (L-826266) and EP(4) (L 161982) receptor antagonists prevented the nociceptive response induced by misoprostol in formalin-injected mice. Prostanoid EP(1), EP(2), EP(3) and EP(4) antagonists reduced, per se, the late hyperalgesic phase. Intra-periaqueductal grey perfusion with misoprostol increased periaqueductal grey glutamate, whereas it produced an increase followed by a decrease in GABA. Likewise, formalin increased glutamate and produced a biphasic response on GABA. When misoprostol was perfused in combination with the peripheral injection of formalin, we observed an increase of glutamate and an increase followed by a stronger decrease in GABA release. These data show that periaqueductal grey prostaglandin receptor stimulation increased formalin-induced nociceptive response in the late phase by increasing glutamate release and by producing a biphasic change in GABA release. PMID- 16360149 TI - Natriuretic peptides differentially attenuate thrombin-induced barrier dysfunction in pulmonary microvascular endothelial cells. AB - Previous studies have described a protective effect of atrial natriuretic peptide (ANP) against agonist-induced permeability in endothelial cells derived from various vascular beds. In the current study, we assessed the effects of the three natriuretic peptides on thrombin-induced barrier dysfunction in rat lung microvascular endothelial cells (LMVEC). Both ANP and brain natriuretic peptide (BNP) attenuated the effect of thrombin on increased endothelial monolayer permeability and significantly enhanced the rate of barrier restoration. C-type natriuretic peptide (CNP) had no effect on the degree of thrombin-induced monolayer permeability, but did enhance the restoration of the endothelial barrier, similar to ANP and BNP. In contrast, the non-guanylyl cyclase-linked natriuretic peptide receptor specific ligand, cyclic-atrial natriuretic factor (c ANF), delayed the rate of barrier restoration following exposure to thrombin. All three natriuretic peptides promoted cGMP production in the endothelial cells; however, 8-bromo-cGMP alone did not significantly affect thrombin modulation of endothelial barrier function. ANP and BNP, but not CNP or c-ANF, blunted thrombin induced RhoA GTPase activation. We conclude that ANP and BNP protect against thrombin-induced barrier dysfunction in the pulmonary microcirculation by a cGMP independent mechanism, possibly by attenuation of RhoA activation. PMID- 16360150 TI - Light-induced exacerbation of retinal degeneration in a rat model of Smith-Lemli Opitz syndrome. AB - Potentiation of retinal degeneration by intense light exposure, and its amelioration by an antioxidant, were studied in a rat model of Smith-Lemli-Opitz syndrome (SLOS), in comparison with normal (control) Sprague-Dawley rats. The SLOS model is created by treating rats with AY9944, a selective inhibitor of cholesterol synthesis at the level of 3beta-hydroxysterol-Delta7-reductase. A subset of rats was treated with dimethylthiourea (DMTU), a synthetic antioxidant, 24 and 1 hr prior to light exposure. Half of the animals (+/-DMTU) were exposed to intense, constant, green light (24hr, 1700lx, 490-580 nm), while the others were maintained in darkness. Subsequently all animals were returned to dim cyclic light (20-40 lx, 12 hr light-12 hr dark) for 2 weeks, after which electroretinograms were recorded. One eye from each rat was taken for histological and quantitative morphometric analyses; sterol analysis was performed on retinas from contralateral eyes. HPLC analysis confirmed the accumulation of 7-dehydrocholesterol (7DHC) in retinas of AY9944-treated rats; cholesterol represented >99% of the sterol in control retinas. Histology of retinas from unexposed, AY9944-treated rats (6-week-old) was normal. In contrast, age-matched, light-exposed rats exhibited massive photoreceptor cell loss in both the superior and inferior hemispheres, and concomitant rod and cone dysfunction. The severity and geographic extent of the damage was far greater than that observed in normal albino rats exposed to the same conditions. DMTU pre-treatment largely prevented these degenerative changes. These findings indicate that the AY9944-induced rat SLOS model is hypersensitive to intense light-induced retinal damage, relative to normal rats. DMTU protection against light-induced damage implicates free radical-based oxidation in the retinal degeneration process. Furthermore, the use of green light (corresponding to the absorption maxima of rhodopsin) implicates rhodopsin in the initiation of the pathobiological mechanism. We propose that generation of cytotoxic oxysterols (by-products of 7DHC oxidation) is an integral part of retinal cell death in the SLOS rat model, which is exacerbated by intense light. Furthermore, the results predict light dependent potentiation of retinal degeneration in SLOS patients, and the possible ameliorative effects of antioxidant therapy. PMID- 16360151 TI - Focus on molecules: keratocan (KERA). PMID- 16360152 TI - Enriched environment induces cellular plasticity in the adult substantia nigra and improves motor behavior function in the 6-OHDA rat model of Parkinson's disease. AB - The adult substantia nigra bears the capacity to generate new neural cells throughout adulthood. The mechanisms of cellular plasticity in this brain region remain unknown. In the adult dentate gyrus, dopamine was suggested to be one of the key players in neurogenesis. We therefore investigated nigral cellular plasticity in the 6-OHDA rat model of Parkinson's disease. The absolute numbers of newborn cells in the SN were not affected by dopamine depletion. Interestingly, we found a specific downregulation of generation of newborn nigral astrocytic cells. As enriched environment with physical activity are robust inducers of neuro- and gliogenesis in the adult DG, we investigated the role of these physiological stimuli in nigral cellular plasticity and in motor behavior of 6-OHDA lesioned rats. We describe a significant increase in numbers of newborn NG2-positive and GFAP-positive cells in the SN. Moreover, 6-OHDA lesioned animals living in enriched environment with physical activity for 7 weeks showed improved motor behavior compared to controls under standard conditions. Thus, physiological neurogenic and gliogenic stimuli induce significant microenvironmental changes in the adult SN and improve motor behavior in the 6 OHDA lesion model of PD. PMID- 16360153 TI - Dimethyl sulfoxide as an inducer of differentiation in preosteoblast MC3T3-E1 cells. AB - Osteoblastic differentiation is an essential part of bone formation. Dimethyl sulfoxide (DMSO) is a water miscible solvent that is used extensively for receptor ligands in osteoblast studies. However, little is known about its effects on osteoblastogenic precursor cells. In this study, we have used a murine preosteoblast cell line MC3T3-E1 cells to demonstrate that DMSO effectively induces osteoblastic differentiation of MC3T3-E1 cells via the activation of Runx2 and osterix and is dependent upon the protein kinase C (PKC) pathways. We further demonstrated that prolonged activation of PKC pathways is sufficient to induce osteoblastic differentiation, possibly via the activation of PKD/PKCmu. PMID- 16360154 TI - Expression of Musashi-1 in the rat stomach and changes during mucosal injury and restitution. AB - Musashi-1 is involved in maintenance of neural stem cells. In the rat stomach, we found some cells in the luminal compartment of the mucosa that stained positively for Musashi-1. These cells were distinct from other cells of epithelial lineage, except for parietal cells, and coexpressed HES5. The Musashi-1-positive cells exfoliated after damage, while Musashi-1 expression in neck cells were upregulated, and proliferating cells diminished before reappearing and increasing in number thereafter. We conclude that a subpopulation of parietal cells acts as a source of Musashi-1, which contributes to rapid re-epithelization by restoration of stem cells and regulation of cell differentiation. PMID- 16360155 TI - Plasma glucocorticoid concentrations and body mass in ground squirrels: seasonal variation and circannual organization. AB - We examined variation in plasma glucocorticoid concentrations of free-living Belding's ground squirrels (Spermophilus beldingi) and captive golden-mantled ground squirrels (Spermophilus lateralis) housed in constant environmental conditions. Plasma corticosterone concentrations were not significantly correlated with time held captive in traps prior to collection of blood samples in males or non-breeding females, but these variables were significantly correlated in breeding females during the pre-mating, lactation, and post lactation periods. Among male S. beldingi, plasma corticosterone concentrations increased over the course of the active season, and were significantly higher in non-breeding than breeding individuals. Corticosterone concentrations also increased in non-breeding females throughout the active period. In breeding females, baseline (non-stress) corticosterone concentrations, determined from blood samples collected within 3 min of capture, increased during gestation and declined during lactation, whereas stress values of corticosterone, determined from blood samples collected within 4-11 min of capture, increased during gestation and then again during the period after young emerged from the natal burrow. Changes in plasma corticosterone concentrations of S. beldingi paralleled changes in body mass. Among S. lateralis, plasma concentrations of cortisol were elevated when males and females were in reproductive condition and lower in reproductively quiescent squirrels. The annual peak in plasma cortisol concentrations occurred just prior to increases in body mass associated with pre hibernation fattening. Collectively, these results suggest that concentrations of circulating glucocorticoids fluctuate with circannual rhythmicity in conjunction with annual cycles of change in body mass and activity; within these circannual cycles glucocorticoid concentrations are influenced by stress and breeding status. PMID- 16360156 TI - Transvaginal sonographic assessment of endometrial thickness and endometrial morphology in postmenopausal bleeding. PMID- 16360157 TI - Occupational stress, job satisfaction, and working environment among Icelandic nurses: a cross-sectional questionnaire survey. AB - BACKGROUND: Nurses' occupational stress decreases job satisfaction, increases turnover rate, and reduces nursing quality. At different workplaces nurses are confronted with different work tasks, working conditions and hence different sources of stress. OBJECTIVES: The purpose of this study was to explore what factors contribute to work-related stress among Icelandic nurses working within and outside the hospital environment. DESIGN: The study used a cross-sectional survey design. SETTINGS AND PARTICIPANTS: The study population was composed of all working nurses registered at the Icelandic Nurses' Association (INA). Approximately 95% (N=2,234) of Icelandic nurses are members of the INA. Questionnaires were posted to 522 (23.4%) randomly selected participants. The response rate was 42% (N=219), representing 9.8% of the population. Data was analyzed from 206 nurses; 35% worked outside the hospital setting and 65% were hospital based. METHODS: Data was gathered on demographic information and indicators of working conditions, occupational stress, workload, and job satisfaction. A stepwise, multiple linear regression model was employed to calculate significant predictors of occupational stress. RESULTS: The findings suggest that the strenuous conditions of Icelandic nurses are felt more severely among hospital nurses than among nurses working outside hospital settings. The study identified which sources of occupational stress are specific to each of the two groups. CONCLUSIONS: The study found several factors that contribute to work related stress. These findings can be used to guide preventive measures to diminish occupational stress among Icelandic nurses. PMID- 16360158 TI - The surveillance of nursing standards: an organisational case study. AB - BACKGROUND: Quality assurance has acquired increasing prominence in contemporary healthcare systems and there has been an 'explosion' of audit activity. Some authors have begun to investigate the impact of audit activity on organisational and professional cultures. This paper considers data from a wider study of the management of the 'problem' nurse. Nurses and managers had contrasting perceptions of the value of different methods of assessing ward standards and their views are presented here. DESIGN AND SETTING: The study involved organisational case studies in three healthcare Trusts in the north of England. The fieldwork for this study was funded by the United Kingdom Central Council for Nursing, Midwifery and Health Visiting under their research scholarship programme. METHODS: Multiple methods were employed including observation, interviewing and documentary analysis. A total of 144 informal interviews were carried out with ward nurses and their managers. FINDINGS: The study demonstrated different viewpoints regarding the surveillance of nursing standards at top management, middle management and ward levels. The paper considers the discrepancies between these different viewpoints. CONCLUSION: None of the participants placed a high value on audit as a method of assessing ward standards. Complaints data and informal methods were more highly valued by managers. Ward nurses stressed the importance of presence and vigilance in assuring high standards of nursing care. PMID- 16360159 TI - Is there more than C-reactive protein and fibrinogen? The prognostic value of soluble CD40 ligand, interleukin-6 and oxidized low-density lipoprotein with respect to coronary and cerebral vascular disease. AB - Incidence of atherosclerosis and atherosclerosis-related complications will increase significantly in the coming decennia. Research identified many serum and plasma markers that are associated with cardiovascular disease. However, little is known about the prognostic value of these markers to identify patients at risk for future cardiovascular events. Therefore, we aimed to investigate the prognostic value of three of these markers (soluble CD40 ligand (sCD40L), interleukin-6 (IL-6) and oxidized low-density lipoprotein (oxLDL)) with respect to coronary vascular disease and stroke. For this reason the Medline database was searched for the period January 1999-January 2005. To be selected in our study, concentration of the marker had to be determined at baseline, follow-up period had to be longer than 3 months and an estimate of relative risk had to be available. Based on these criteria, 4 studies for sCD40L, 10 for IL-6 and 2 for oxLDL were selected. Relative risk estimates adjusted for potential confounders varied between 1.9 and 2.8 for sCD40L, between 1.1 and 3.1 for IL-6 and between 1.9 and 3.2 for oxLDL. In conclusion, this systematic review shows that sCD40L, IL-6 and oxLDL are associated with an increased relative risk of developing cardiovascular disease. PMID- 16360160 TI - The flexural rigidity of the aortic valve leaflet in the commissural region. AB - Flexure is a major deformation mode of the aortic valve (AV) leaflet, particularly in the commissural region where the upper portion of the leaflet joins the aortic root. However, there are no existing data known on the mechanical properties of leaflet in the commissural region. To address this issue, we quantified the effective stiffness of the commissural region using a cantilever beam method. Ten specimens were prepared, with each specimen flexed in the direction of natural leaflet motion (forward) and against the natural motion (reverse). At a flexure angle (phi) of 30 degrees , the effective forward direction modulus E was 42.63+/-4.44 kPa and the reverse direction E was 75.01+/ 14.53 kPa (p=0.049). Further, E-phi response was linear (r(2) approximately 0.9) in both flexural directions. Values for dE/dphi were -2.24+/-0.6 kPa/ degrees and -1.90+/-0.3 kPa/ degrees in the forward and reverse directions, respectively (not statistically different, p=0.424), indicating a consistent decrease in stiffness with increased flexure. In comparison, we have reported that the effective tissue stiffness of AV leaflet belly region was 150-200 kPa [Merryman, W.D., Huang, H.Y.S., Schoen, F.J., Sacks, M.S. (2006). The effects of cellular contraction on AV leaflet flexural stiffness. Journal of Biomechanics 39 (1), 88-96], which was also independent of direction and amount of flexure. Histological studies of the commissure region indicated that tissue buckling was a probable mechanism for decrease in E with increasing flexure. The observed change in E with flexural angle in the commissural region is a subtle aspect of valve function. From a valve design perspective, these findings can be used as design criteria in fabricating prosthetic devices AV resulting in better functional performance. PMID- 16360161 TI - Time dependent properties of bovine meniscal attachments: stress relaxation and creep. AB - It has been suggested that the success of a meniscal replacement is dependent on several factors, one of which is the secure fixation and firm attachment of the replacement to the tibial plateau [Chen, M.I., Branch, T.P., et al., 1996. Is it important to secure the horns during lateral meniscal transplantation? A cadaveric study. Arthroscopy 12(2), 174-181; Alhalki, M.M., et al., 1999. How three methods for fixing a medial meniscal autograft affect tibial contact mechanics. American Journal of Sports Medicine 27(3), 320-328; Haut Donahue, T.L., et al., 2003. How the stiffness of meniscal attachments and meniscal material properties affect tibio-femoral contact pressure computed using a validated finite element model of the human knee joint. Journal of Biomechanics 36(1), 19-34]. The complex loading environment in the knee lends itself to different loading environments for each meniscal attachment. We hypothesize that the creep and stress relaxation characteristics of the horn attachments will be different for the anterior versus posterior, and medial versus lateral attachments. To test this hypothesis, the stress relaxation and creep characteristics of the meniscal horn attachments were determined. The stress relaxation properties of load/stress at the end of the test, and the load/stress relaxation rate demonstrated no significant statistical differences between the attachments. Unlike the stress relaxation properties, the creep properties demonstrated some significant differences amongst the attachments. The normalized displacement at the end of the test, normalized creep rate and strain creep rate for the lateral anterior attachment were significantly different than those of the medial posterior attachment (p<0.05). The two anterior attachments had significantly different strains at the end of the test, as well as significantly different creep strain rates (p<0.05). The two attachments of the medial meniscus revealed no significant differences between any of the creep properties measured (p>0.05). The time dependent properties obtained in this experiment provide insight into the behavior of meniscal horn attachments under various loading situations. The results indicate that a suitable meniscal replacement may require different properties for the lateral and medial horns. PMID- 16360162 TI - Metathesis polymerization-derived monolithic membranes for solid-phase extraction coupled with diffuse reflectance spectroscopy. AB - Novel monolithic disks were prepared via ring opening metathesis polymerization (ROMP) from norborn-2-ene (NBE), a crosslinker, i.e., 1,4,5,8,8a-hexahydro 1,4,4,5,8, exo, endo-dimethanonaphthalene (DMN-H6) and tris(norborn-5-ene-2 ylmethylenoxy)methylsilane, respectively, 2-propanol and toluene (25:25:41:9, all %, w/w) using RuCl2(PCy3)2(CHPh) (Cy=cyclohexyl) as initiator and triphenylphosphine (PPh3) as modulator. Disks 1-2 mm thick were prepared by polymerization in a mold, disks thinner than 1mm were prepared by impregnation of nylon or other porous filters prior to the polymerization step. These disks were evaluated for the preconcentration of iodine and selected organic solutes from dilute aqueous samples by solid-phase extraction (SPE). Quantitative measurement of the extracted solutes was achieved by diffuse-reflectance spectroscopy (DRS) directly on the surface of the disk. PMID- 16360163 TI - Detailed characterization of cationic hydroxyethylcellulose derivatives using aqueous size-exclusion chromatography with on-line triple detection. AB - A more complete understanding of polymeric, cationic cellulose derivatives, including polyquaterium-10 (Polymer JR), has become increasingly important in the eye care industry as thorough characterization of raw materials helps promote product quality and process control. Often such detailed information requires utilization of a combination of analytical techniques. In this work three Polymer JR samples with different viscosities were characterized using aqueous size exclusion chromatography (SEC) with a light scattering detector, a differential viscometer, and a differential refractometer (triple detection). Detailed molecular information such as absolute molecular weights, molecular weight distributions, intrinsic viscosities, and molecular conformations were obtained. One major challenge of analyzing cationic polymers is abnormal size exclusion separation, which could be caused by the ionic interaction between sample molecules and the column packing material. A selection of mobile phases varying in pH, buffer, organic solvent content, and molar concentration of salts was employed to evaluate the correlation of obtained molecular weight values and mobile phase composition. Universal calibration concept was used to examine the abnormal size exclusion separation phenomenon of Polymer JR samples when using different mobile phases. It was observed that the abnormal size exclusion was dependent on both the separation conditions and molecular weights of the samples. Despite the changes in separation parameters and uncharacteristic polymeric structure compared to conventional SEC samples, the use of aqueous SEC with triple detection provided reproducible and valuable molecular information of Polymer JR samples with low to medium molecular weights. By using a combination of high buffer content and adding organic solvent, the abnormal exclusion separation of high molecular weigh Polymer JR could be considerably reduced. PMID- 16360164 TI - New approach on similarity analysis of chromatographic fingerprint of herbal medicine. AB - A new approach to the construction and similarity analysis of chromatographic fingerprint for herbal medicine is presented in this paper. Samples of chuanxiong, a herbal medicine for headache, from three producing areas of China were used to evaluate the utility of this study. The samples were analyzed with high-performance liquid chromatography (HPLC) and the peak areas of the chromatograms were used to construct the fingerprints of the herbal medicines. A vector of differences was defined between the two fingerprints. The scalar mean of the difference vector was taken as a statistic and both the t-test and Bayesian hypothesis testing were implemented to provide a one-to-one comparison of the fingerprints. Compared with principal component analysis (PCA), correlation coefficient and vector cosine, the new method offers a better differentiation of the similarity or difference between the fingerprints from same sample of chuanxiong. When the new method was used in the similarity analysis of the fingerprints of chuanxiong from different production areas, a clear-cut signature was obtained that reveals the significant difference between them. PMID- 16360165 TI - The effect of visual word features on the acquisition of orthographic knowledge. AB - Research with adults has shown that the distortion of visual word features, and in particular of the multiletter features within words, hampers word recognition. In this study, "CaSe MiXiNg" was employed to examine the effect of disrupting visual word features on the acquisition of orthographic knowledge in children. During the training, 18 beginning and 27 advanced readers (in Grades 2, 4, and 5) repeatedly read a set of pseudowords in either lowercase or mixed case. During this training, case mixing appeared to impair reading speed in both reader groups. At posttest, 1 day after the training, case format was either the same as or different from that during the training. Lowercase pseudowords were recognized faster after a lowercase training than after a mixed-case training. In a second study, case was found not to affect the rapid naming of single letters. The combined results suggest that case mixing disrupted the multiletter features in pseudowords and that the disruption of these features can affect the acquisition of orthographic knowledge. PMID- 16360166 TI - What counts as knowing? The development of conceptual and procedural knowledge of counting from kindergarten through Grade 2. AB - The development of conceptual and procedural knowledge about counting was explored for children in kindergarten, Grade 1, and Grade 2 (N = 255). Conceptual knowledge was assessed by asking children to make judgments about three types of counts modeled by an animated frog: standard (correct) left-to-right counts, incorrect counts, and unusual counts. On incorrect counts, the frog violated the word-object correspondence principle. On unusual counts, the frog violated a conventional but inessential feature of counting, for example, starting in the middle of the array of objects. Procedural knowledge was assessed using speed and accuracy in counting objects. The patterns of change for procedural knowledge and conceptual knowledge were different. Counting speed and accuracy (procedural knowledge) improved with grade. In contrast, there was a curvilinear relation between conceptual knowledge and grade that was further moderated by children's numeration skills (as measured by a standardized test); the most skilled children gradually increased their acceptance of unusual counts over grade, whereas the least skilled children decreased their acceptance of these counts. These results have implications for studying conceptual and procedural knowledge about mathematics. PMID- 16360167 TI - The eyes of a patrolling butterfly: visual field and eye structure in the Orange Sulphur, Colias eurytheme (Lepidoptera, Pieridae). AB - Sensory information plays a critical role in determining an animal's behavior on both proximate and evolutionary timescales. Butterflies, like many other insects, use vision extensively over their lifetimes, and yet relatively little work has been published to date on their visual capabilities. We describe the visual system of a pierid butterfly, Colias eurytheme, with the ultimate goal of better understanding its role in shaping the behavior of this animal. We made several measurements: visual field dimensions, eye surface area, interommatidial angle (Deltaphi), facet diameter (D), and eye parameter (p). C. eurytheme had a large visual field and considerable regional variation in visual acuity, as inferred by Deltaphi and D. When compared to females, males had larger eye surface areas, smaller Deltaphi, and larger D in all regions except ventrally. Both sexes had proportionally large eye surface areas compared to other butterflies. Minimum p in males was small, indicating that some regions of their eyes may operate close to the diffraction limit. Finally, we found that both eye surface area and D scaled positively, but with negative allometry to body size. We discuss the relevance of these visual characteristics to the biology and behavior of C. eurytheme. PMID- 16360168 TI - cDNA cloning and in situ hybridization of Delta11-desaturase, a key enzyme of pheromone biosynthesis in Ostrinia scapulalis (Lepidoptera: Crambidae). AB - Female sex pheromones are considered to be produced in a "pheromone gland" located in the terminal abdominal segments (8th-10th, TAS) of a moth; however, in many moth species, the cells that produce pheromones have not actually been specified. We investigated cells in the TAS that synthesize pheromones in the adzuki bean borer Ostrinia scapulalis, by locating pheromones and their precursors, and mRNA for Delta11-desaturase, a key enzyme in pheromone biosynthesis. We demonstrated that the pheromone components, (E)-11- and (Z)-11 tetradecenyl acetates, and their fatty acyl precursors were specifically contained in the dorsal part of the TAS. A cDNA (OscaZ/E11) that encodes a Delta11-desaturase was cloned from the TAS. RT-PCR and in situ hybridization unequivocally showed that OscaZ/E11 is specifically expressed in the modified epidermal cells located at the dorsal end of the 8th-9th intersegmental membrane. PMID- 16360169 TI - High-resolution crystal structure of a truncated ColE7 translocation domain: implications for colicin transport across membranes. AB - ColE7 is a nuclease-type colicin released from Escherichia coli to kill sensitive bacterial cells by degrading the nucleic acid molecules in their cytoplasm. ColE7 is classified as one of the group A colicins, since the N-terminal translocation domain (T-domain) of the nuclease-type colicins interact with specific membrane bound or periplasmic Tol proteins during protein import. Here, we show that if the N-terminal tail of ColE7 is deleted, ColE7 (residues 63-576) loses its bactericidal activity against E.coli. Moreover, TolB protein interacts directly with the T-domain of ColE7 (residues 1-316), but not with the N-terminal deleted T-domain (residues 60-316), as detected by co-immunoprecipitation experiments, confirming that the N-terminal tail is required for ColE7 interactions with TolB. The crystal structure of the N-terminal tail deleted ColE7 T-domain was determined by the multi-wavelength anomalous dispersion method at a resolution of 1.7 angstroms. The structure of the ColE7 T-domain superimposes well with the T domain of ColE3 and TR-domain of ColB, a group A Tol-dependent colicin and a group B TonB-dependent colicin, respectively. The structural resemblance of group A and B colicins implies that the two groups of colicins may share a mechanistic connection during cellular import. PMID- 16360170 TI - Cigarette smoking in Chinese male inpatients with schizophrenia: a cross sectional analysis. AB - Given the high rates of cigarette smoking in schizophrenia in many published studies from around the world, we studied the relationship between smoking status and clinical characteristics in male Chinese schizophrenic inpatients. Two hundred seventy-nine schizophrenic inpatients were assessed using clinical data forms to ascertain historical, demographic and treatment variables and collateral information was also collected from case records and interviews with patients and family members. Current smokers (N=112) were compared with non-smokers (N=167) on clinical variables by independent sample t-tests and chi(2) tests, with adjustment for confounding variables using ANCOVA and binary logistic regression analysis. Compared to non-smokers, current smokers were significantly more likely to be divorced, have lower educational attainment, a more episodic course, have a greater number of previous psychotic relapses and more likely to be treated with clozapine. There was no correlation, however, between smoking consumption and schizophrenic psychopathology. Accordingly, cigarette smoking may be associated with certain clinical features in schizophrenic patients, and should be carefully screened for when making treatment and rehabilitation plans. PMID- 16360171 TI - Exocrine meets endocrine: pancreatic stone protein and regenerating protein--two sides of the same coin. AB - BACKGROUND: Regenerating protein (reg) and pancreatic stone protein (PSP) have been discovered independently in the fields of diabetes and pancreatitis. MATERIALS AND METHODS: These proteins are identical; however, because of the gap between the endocrine and exocrine field, there was never a consensus and the nomenclature has not been rectified. Since the time of the initial discovery, more isoforms have been unified. Historically, PSP was discovered long before reg, yet, in many areas outside of the pancreatitis research field, reg is being used. RESULTS: For PSP/reg, a role in proliferation and regeneration of islet cells has been postulated. A hitherto insufficiently understood phenomenon is the massive up-regulation of PSP/reg in pancreatic tissue and juice under conditions of stress. Similarly, PAP (pancreatitis-associated protein)/reg III has been attributed various functional roles. Structurally, the ability to form fibrils after tryptic cleavage is a striking common features of both proteins. However, this biochemical transformation is in itself not enough to gain functional insight. Thus, physiological and genetic approaches are required to further characterize the role of these proteins in the pancreas. Recently, more evidence has been presented in support of the theory that PSP/reg plays a key role in islet neogenesis/regeneration. CONCLUSIONS: In this review we discuss the debate on the localization and functional roles of PSP/reg and PAP/regIII. Therefore, we have summarized hypotheses and experimental results supporting such hypotheses. PMID- 16360172 TI - Nitrotyrosine and chlorotyrosine: clinical significance and biological functions in the vascular system. AB - The heme-containing enzyme myeloperoxidase (MPO) is both present and active in inflammatory conditions. This enzyme is potentiated by its formation of multiple inflammatory mediators. The two most common mediators are the modified tyrosines: nitrotyrosine and 3-chlorotyrosine. Along with other modified tyrosines, these molecules have been found to be elevated in atherosclerosis, lung disease, sepsis, vasculitis, and other inflammatory diseases. By treating some of these diseases, the levels of modified tyrosines have been shown to decrease. There have been a wide range of animal models designed to study the in vivo effects of these tyrosine molecules. In addition, there are also several reports in the literature of the in vitro actions of modified tyrosine molecules demonstrated by various cell-culture models. The purpose of this review is to evaluate the clinical significance and biological functions of these modified tyrosine molecules in atherosclerosis as well as a variety of other inflammatory conditions. It is timely information because of their association with diseases as well as lack of overview of their molecular actions. This review will focus on the formation, clinical significance, and animal and cell-culture models of these important molecules. PMID- 16360173 TI - Salicylate enhances rat gastric gelatinase activity. AB - BACKGROUND: Increased matrix metalloproteinase (MMP) activity is associated with tissue injury in some organs. Their role in gut injury remains to be fully elucidated. We recently demonstrated that increased MMP-2 activity participated in lipopolysaccharide (LPS)-induced gastric injury. Thus we hypothesized that MMPs may play a role in other models of gastric injury. MATERIALS AND METHODS: The effect of L-NAME (10 mg/kg IP) or salicylate (100 mg/kg IP) on gastric injury from 20% ethanol was evaluated in an anesthetized model of gastric injury. In a separate experiment, gastric metalloproteinase activity was assessed after salicylate or L-NAME administration. Rats were given either L-NAME (10 mg/kg), salicylate (100 mg/kg), or saline IP and sacrificed after 6 hours. Gastric mucosa was harvested and portions of the glandular stomach snap frozen for gelatin and in situ zymography as indices of MMP activity. Subsequently the effect of MMP inhibition on macroscopic gastric injury from salicylate and a dilute luminal irritant was determined. RESULTS: Both L-NAME and salicylate significantly increased gastric injury from 20% ethanol versus saline controls. Salicylate treatment significantly increased gelatinase activity as determined by in situ zymography and gelatin zymography while L-NAME did not. MMP inhibition ameliorated macroscopic gastric injury secondary to salicylate and a dilute luminal irritant. CONCLUSIONS: This is the first study to report that MMP activity increases in the stomach following salicylate treatment. These data suggest that MMPs may play a role in the ability of salicylate to exacerbate gastric injury from irritants, but likely do not play a role in mediating the deleterious effects of L-NAME. PMID- 16360174 TI - Cold preservation of islets in UW solution--with special reference to apoptosis. AB - BACKGROUND: Apoptosis progresses in cultured islets. Little is known with regard to apoptosis under cold preservation. We examined viability and function of islets in University of Wisconsin (UW) solution. MATERIALS AND METHODS: Isolated rat islets were cultured overnight (overnight group) and further treated with 7 day culture in RPMI 1640 medium at 37 degrees C (culture group) or 7-day preservation in UW solution at 4 degrees C (preservation group). They were evaluated by glucose-stimulated insulin secretion test. Apoptosis was examined by TdT-mediated dUTP-biotin nick end-labeling (TUNEL) assay. Expression of caspase mRNA and the ratio of Bax to Bcl-2 were evaluated by reverse-transcriptase polymerase chain reaction (RT-PCR). RESULTS: Islet recovery after 7 days was significantly lower in culture group than in preservation group (44.0 +/- 3.7% versus 75.0 +/- 4.9%, P < 0.05). The stimulation index in the culture group was significantly lower than in the overnight group (2.1 +/- 0.2 versus 4.1 +/- 0.4, P < 0.05). The apoptotic index in the culture group was significantly higher than both in the overnight group and in the preservation group (38.0 +/- 3.0% versus 10.8 +/- 2.0 and 27.0 +/- 4.0%, P < 0.05). Caspase 3, 8, and 9 mRNA in the culture group expressed more than in the other groups. Bax/Bcl-2 in the culture group was significantly lower than in the overnight group (3.2 +/- 0.66 versus 8.1 +/- 0.95, P < 0.05), suggesting that apoptosis had been already destined early after isolation. CONCLUSIONS: The preservation group showed better recovery and function than the culture group. Apoptosis contributed to islet loss under culture and it was significantly suppressed under cold preservation. PMID- 16360175 TI - Does CO2 pneumoperitoneum alter the ultra-structuture of the mesothelium? AB - BACKGROUND: Pneumoperitoneum may be responsible for ultra-structural alterations in the mesothelium during laparoscopy. To characterize the effect of pneumoperitoneum on the mesothelial cells with CO(2) and compressed air; and to compare to laparotomy and control group (anesthesia only). MATERIALS AND METHODS: Forty C-57 mice were divided in four groups of 10 animals each: CO(2), air, laparotomy, and control group. The animals were submitted to pneumoperitoneum at 8 mmHg during 30 min (CO(2) or compressed air). Five animals of each group were sacrificed 2 and 24 h after the procedure. Fragments of parietal peritoneum were collected and processed for scanning electron microscopy. RESULTS: Control group revealed uninterrupted mesothelial cells, without any evidence of cellular limits; close contact between the cells; absence of intercellular clefts and presence of microvilli. In the laparotomy group, similar results to the control group, with decreased microvilli were noted. Air pneumoperitoneum was associated with alterations in the morphology of the mesothelial cells, clear cellular limits, and cells with spherical and fusiforme formats. CO(2) pneumoperitoneum showed mesothelial cells with clear cellular limits, predominantly spherical cellular format, and intercellular clefts that allowed the visualization of the exposed basal membrane. These alterations were more intense after 24 h. There was a statistical significance between CO(2) group (2 and 24 h) compared to the control group and laparotomy for cellular limits, intercellular clefts and microvilli, P < 0.0001. CONCLUSIONS: Pneumoperitoneum causes damage in the mesothelial ultra-structure, which differs from the laparotomy group. CO(2) pneumoperitoneum is more harmful to the mesothelium than the air. PMID- 16360176 TI - In-vivo evaluation of a novel bipolar radiofrequency device for interstitial thermotherapy of liver tumors during normal and interrupted hepatic perfusion. AB - BACKGROUND: Only monopolar systems have thus far been available for radiofrequency ablation of liver tumors, whose application is restricted because of the incalculable energy flow, reduction of electrical tissue conduction, and limited lesion size. The aim of this study was to evaluate a novel internally cooled bipolar radiofrequency application device under in vivo conditions and to compare the effect of this system on lesion size when combined with hepatic arterial microembolization or complete hepatic blood flow occlusion. MATERIALS AND METHODS: In a porcine liver model, RFA (60 W, 12 min) was performed with either normal (n = 12), partially interrupted (arterial microembolization via a hepatic artery catheter n = 12) or completely interrupted hepatic perfusion (Pringle's maneuver, n = 12). RFA parameters (impedance, power output, temperature, applied energy) were determined continuously during therapy. RFA lesions were macroscopically assessed after liver dissection. RESULTS: Bipolar RFA induced clinical relevant ellipsoid thermal lesions without complications. Hepatic inflow occlusion led to a 4.3-fold increase in lesion volume after arterial microembolization and a 5.8-fold increase after complete interruption (7.4 cm(3)versus 31.9 cm(3)versus 42.6 cm(3), P < 0.01). CONCLUSIONS: The novel bipolar RFA device is a safe and effective alternative to monopolar RFA-systems. Interrupting hepatic perfusion significantly increases lesion volumes in bipolar RFA. This beneficial effect can also be achieved in the percutaneous application mode by RFA combined with arterial microembolization via a hepatic artery catheter. PMID- 16360177 TI - Ethanol induction of complete heart block in swine. AB - OBJECTIVES: A method for the induction of complete heart block (CHB) by ethanol injection and its success rate in a pig model of acute right ventricular failure is reported. Additionally, a review of the literature for the induction of CHB in laboratory animals is detailed. The literature review was undertaken to both compare our rate of success with other methods and provide insight into our technique and refine its implementation. BACKGROUND: Animal models of CHB have facilitated the understanding of therapeutics for various cardiac pathologies in humans. In our laboratory, CHB in pigs is used for complete control of heart rhythm in studies of biventricular pacing. MATERIALS AND METHODS: Experiments carried out on pigs in our laboratory that required the induction of CHB were reviewed retrospectively. In addition, review of the literature for creating CHB in animals was undertaken. Our success rate was compared to that of other groups. RESULTS: Our success rate (93%) is similar to other models of CHB, in general, and to those models that used the injection of caustic substances with thoracotomy. CONCLUSIONS: Review of the literature indicates that our success rate is comparable to other groups and that, although many approaches have been described in both open- and closed-chest models, success is likely dependent on the practice and skill of the experimenter. In addition, review of the literature has afforded us new perspectives on the experimental induction of CHB. PMID- 16360179 TI - Selection for high gamete encounter rates explains the evolution of anisogamy using plausible assumptions about size relationships of swimming speed and duration. AB - A previous general model describing physical constraints on gamete encounter rates was modified to incorporate assumptions that increased size causes decreased swimming speed and increased fertile period (or other proportional enhancement to gamete fertility). The analysis indicates that with moderately strong size dependence of fertile period and a range of speed dependencies, selection for high encounter rates pressures mating systems that develop any heritable difference in size between the gametes of different mating types to exaggerate the difference and evolve from isogamy to anisogamy. The smaller gamete has an optimal size, but the larger faces continuing selection for increased size. This continues to a size that is estimated to be sufficient to make pheromone production of sperm attractants practical. This mechanism then bridges the missing link between isogametes and oogamy in a previous analysis of the effectiveness of pheromones in explaining the success of male-female mating systems. The evolution and success of anisogamy and oogamy can be explained solely on the basis of physical effects on the encounter process. PMID- 16360178 TI - Characterization of growth and osteogenic differentiation of rabbit bone marrow stromal cells. AB - BACKGROUND: The rabbit is recognized as an excellent model to study the repair of bony defects with tissue engineered constructs. However, the use of rabbit bone marrow stromal cells (RBMSCs) has been limited despite the proven benefits of autologous BMSCs in the formation of bone. The purpose of this study was to characterize the growth and differentiation pattern of RBMSCs and their response to growth factors. MATERIAL AND METHODS: BMSCs were isolated from New Zealand White rabbits and cultured. Serial cell counts of parallel cultures were taken daily to determine cell growth. Response of RBMSCs to varying doses of recombinant human BMP-2 (rhBMP-2) and their time course was gauged by alkaline phosphatase (ALP) activity. The osteoblastic differentiation potential of RBMSCs in response to rhBMP-2 treatment was determined by evaluating the expression pattern of various genes involved with osteogensis using northern analysis. Von Kossa staining was performed to determine the effect of rhBMP-2 on the mineralization capabilities of RBMSCs. RESULTS: The growth rate of RBMSCs severely declined after first passage and this rate was further suppressed by TGF beta1. The optimal dose response of rhBMP-2 was determined to be 50 ng/ml and its time course displayed increasing alkaline phosphatase activity over time. Two osteogenic markers, collagen I and osteopontin, were up regulated by rhBMP-2 treatment. Finally, the mineralization capability of RBMSCs was determined to be enhanced by rhBMP-2 treatment. CONCLUSION: Our work indicates that RBMSCs possess strong osteogenic potential and can be successfully applied toward bone tissue engineering in a rabbit model. PMID- 16360180 TI - Spatial self-organization in a cyclic resource-species model. AB - Biological communities are remarkable in their ability to form cooperative ensembles that lead to coexistence through various types of niche partitioning, usually intimately tied to spatial structure. This is especially true in microbial settings where differential expression and regulation of genes allows members of a given species to alter their lifestyle so as to fill a functional role within the community. The resulting species interactions can involve feedback, as in the case of some bacterial consortia that participate in the cooperative degradation of a given resource in a succession of steps and in such a way that certain "later" species provide catalytic support for the primary degrader. We seek to capture the essential features of such spatially extended biological systems by introducing a lattice-based stochastic spatial model (interacting particle system) with cyclic local dynamics. Here, a given site progresses through a sequence of resource and species states in a prescribed order. Furthermore, this succession of states (at a site) is assumed to form a cyclic pattern due to a natural feedback mechanism. We explore conditions under which all the species are able to coexist and consider the extent to which this coexistence requires the development of spatio-temporal patterns, including spiral waves. This self-organization, if it occurs, results when synchronization of the dynamics at the microscopic level leads to macroscopic patterns. These patterns result in consumer-driven resource fluctuations that generate a form of spatio-temporal niche partitioning. As with most models of this complexity, we employ a mixture of mathematical analysis and simulations to develop an understanding of the resulting dynamics. PMID- 16360181 TI - Ca2+ signal stimulates the expression of steroidogenic acute regulatory protein and steroidogenesis in bovine adrenal fasciculata-reticularis cells. AB - Adrenal glucocorticoid synthesis is stimulated by ACTH or its nitrophenylsulphenyl derivative, NPS-ACTH. Acute stimulation of steroid hormone biosynthesis is highly dependent on the expression of steroidogenic acute regulatory (StAR) protein. To determine the regulatory mechanism of StAR expression in bovine fasciculata/reticularis cells, we analyzed the second messenger systems involved in StAR protein expression using cultured cells activated by ACTH and NPS-ACTH. We concluded that cAMP is not the essential second messenger for StAR protein expression, since NPS-ACTH activated StAR protein expression more than ACTH without increase in cellular cAMP. A 15 lipoxygenase metabolite(s) of arachidonic acid stimulated steroidogenesis without increase in StAR protein expression, since AA-861, a lipoxygenase inhibitor, inhibited steroidogenesis without affecting StAR protein expression. Stimulation of StAR protein expression and the corresponding increase in the steroidogenesis were inhibited by nicardipine in cells treated with ACTH or NPS-ACTH. These data indicate that the dominant second messenger for the stimulation of StAR protein expression is Ca2+. Calmodulin-dependent kinase II inhibitors KN-93 and KN-62 suppressed steroidogenic activity without affecting StAR expression. The protein kinase C inhibitor Ro 31-8220 did not show any effects on StAR expression and steroidogenesis. Calmodulin-dependent kinase II and protein kinase C can therefore be concluded not to be involved in StAR protein expression in bovine cells. PMID- 16360182 TI - Expression of opioid receptors and c-fos in CB1 knockout mice exposed to neuropathic pain. AB - The development of neuropathic pain is associated with multiple changes in gene expression occurring in the dorsal root ganglia (DRG) and spinal cord. The goal of this study was to evaluate whether the disruption of CB1 cannabinoid receptor gene modulates the changes induced by neuropathic pain in the expression of mu- (MOR), delta- (DOR) and kappa-opioid receptors (KOR) mRNA levels in the DRG and spinal cord. The induction of c-fos expression in the lumbar and sacral regions of the spinal cord was also evaluated in these animals. Opioid receptors mRNA levels were determined by using real-time PCR and Fos protein levels by immunohistochemistry. Nerve injury significantly reduced the expression of MOR in the DRG and the lumbar section of the spinal cord from CB1 cannabinoid knockout (KO) mice and wild-type littermates (WT). In contrast, mRNA levels of DOR and KOR were not significantly changed in any of the different sections analysed. Furthermore, sciatic nerve injury evoked a similar increase of c-fos expression in lumbar and sacral regions of the spinal cord of both KO and WT. In all instances, no significant differences were observed between WT and KO mice. These data revealed specific changes induced by neuropathic pain in MOR expression and c-fos levels in the DRG and/or spinal cord that were not modified by the genetic disruption of CB1 cannabinoid receptors. PMID- 16360183 TI - Molecular analysis of the interaction of the pyrazine derivatives RPR119990 and RPR117824 with human AMPA-type glutamate receptor channels. AB - Antagonizing glutamatergic neurotransmission by blockade of AMPA-type glutamate receptors is a promising pharmacological strategy in the treatment of neurodegenerative diseases. We investigated the interaction of two new pyrazine derivatives (RPR119990 and RPR117824) with recombinant AMPA-type glutamate receptors. Recombinant homooligomeric GluR1flop, GluR2flip, GluR2flop, GluR6, non desensitizing GluR2 L504Y channels and heterooligomeric GluR1/2 channels were expressed in HEK293 cells. AMPA-type channels were competitively inhibited by RPR119990 or RPR117824 with an IC(50) around 10 nM, at GluR6 channels the dose response relation of the inhibition was shifted to higher concentrations. Non desensitizing GluR2 L504Y channels were used to further characterize the inhibition. After equilibration with the agonist a marked dose-dependent current decay upon coapplication of glutamate and RPR119990 and a dose-independent time course of recovery from block was observed. The extents of current inhibition as well as the time constant of current decay upon addition of the blocker to the test solution were dependent on agonist concentration, pointing to a competitive inhibition. Quantitative analysis of the experimental data using computerized simulations are compatible with a competitive block mechanism and provides hints to binding sites at unliganded and liganded closed states of the receptor. PMID- 16360184 TI - Counting in everyday life: discrimination and enumeration. AB - Enumerating the number of items in a set accurately and quickly is a basic mathematical skill. This ability is especially crucial in the more real-life situations, where relevant items have to be discriminated from irrelevant distracters. Although much work has been done on the brain mechanisms and neural correlates of the enumeration and/or discrimination process, no agreement has been reached yet. We used event-related potentials (ERPs) to show the time course of brain activity elicited by a task that involved both enumeration and discrimination at the same time. We found that even though the two processes run to some extent in parallel, discrimination seems to take place mainly in an earlier time window (from 100 ms after the stimulus onset) than enumeration (beyond 200 ms after the stimulus onset). Moreover, electrophysiological evidence based on the N2 and P3 components make it reasonable to argue for the existence of a dichotomy between subitizing (for sets of less than four items) and counting (for sets of four and more items). Source estimation suggests that subitizing and counting, though being distinct brain processes, do recruit similar brain areas. PMID- 16360185 TI - Sleep is increased in mice with obesity induced by high-fat food. AB - Excessive daytime sleepiness has been associated with obesity in humans. However, experimental studies on sleep in obese animals are scarce and the results are not consistent. To test the hypothesis that obesity is associated with increased sleep, we examined the effects of obesity, induced by high-fat food, on sleep in mice. We first determined baseline sleep in adult C57BL/6 mice (6 months of age). In the following 6 weeks, the experimental mice (n = 12) were switched to high fat food, in which fat provided 59% of calories, and the control mice (n = 11) were continuously fed with regular lab chows, in which fat provided 16% of calories. The body weights increased steadily in the high-fat group, but maintained constant in the controls. Wakefulness was reduced when assessed after 2, 4, and 6 weeks of high-fat feeding. Concurrently, there were large increases (about 80-100 min/day) in non-rapid eye movement sleep (NREMS). Rapid eye movement sleep (REMS) was not altered. The numbers of NREMS and REMS episodes were increased, whereas the duration of waking episodes was reduced, mainly during the dark period. These alterations in sleep were not observed in the controls. In the high-fat group, the increases of body weight, but not the amounts of energy intake, were negatively correlated with the change in the amounts of wakefulness and positively correlated with the change in the amounts of NREMS. These results indicate that the obese animals have increased sleep pressure and difficulties in maintaining wakefulness during the active phase. PMID- 16360186 TI - Effects of stress on parental care are sexually dimorphic in prairie voles. AB - The effects of stress on parental care are poorly understood, especially in biparental species where males also display care. Data from previous studies in prairie voles, as well as parallels with pair-bonding behavior, suggest the hypothesis that a stressful experience might facilitate parental care in males but not in females. In the present study, male and female prairie voles were exposed to either a 3-min swim stressor or no stressor; 45 min later each animal was tested in a parental care paradigm. Following the parental care test, blood samples were collected and assayed for corticosterone (CORT). After the stressor males, but not females, showed significant changes in parental behavior including significantly more time in kyphosis (arched-back huddling), and a tendency to spend more time licking and grooming pups. In males, CORT levels measured following the parental care test were inversely related to licking and grooming but positively correlated with retrievals. These findings support earlier studies suggesting that the neuroendocrine substrates of parental behavior, as well as the effects of stressors, are sexually dimorphic in this species. PMID- 16360187 TI - The dynamic effects of an inducible defense in the Nicholson-Bailey model. AB - We investigate the dynamic effects of an inducible prey defense in the Nicholson Bailey predator-prey model. We assume that the defense is of all-or-nothing type but that the probability for a prey individual to express the defended phenotype increases gradually with predator density. Compared to a defense that is independent of predation risk, an inducible defense facilitates persistence of the predator-prey system. In particular, inducibility reduces the minimal strength of the defense required for persistence. It also promotes stability by damping predator-prey cycles, but there are exceptions to this result: first, a strong inducible defense leads to the existence of multiple equilibria, and sometimes, to the destruction of stable equilibria. Second, a fast increase in the proportion of defended prey can create predator-prey cycles as the result of an over-compensating negative feedback. Non-equilibrium dynamics of the model are extremely complex. PMID- 16360188 TI - Prediction of effects of genetic drift on variance components under a general model of epistasis. AB - For a model of diallelic loci with arbitrary epistasis, Barton and Turelli [2004. Effects of genetic drift on variance components under a general model of epistasis. Evolution 58, 2111-2132] gave results for variances among and within replicate lines obtained by inbreeding without selection. Here, we discuss the relation between their population genetic methods and classical quantitative genetic arguments. In particular, we consider the case of no dominance using classical identity by descent arguments, which generalizes their results from two alleles to multiple alleles. To clarify the connections between the alternative methods, we obtain the same results using an intermediate method, which explicitly identifies the statistical effects of sets of loci. We also discuss the effects of population bottlenecks on covariances among relatives. PMID- 16360189 TI - Characterization of biofilm structure and its effect on membrane permeability in MBR for dye wastewater treatment. AB - Two membrane bioreactors were operated at aerobic (DO=6.0mg/L) and anoxic (DO<0.3mg/L) conditions for the treatment of synthetic dye wastewater to determine the effect of dissolved oxygen on membrane filterability. The rate of membrane fouling for the anoxic MBR was five times faster than that for the aerobic MBR. Differences in the nature of the biofilm that was formed on the membrane surface as the result of different DO level was the main factor in the different fouling rates. The biofilm structure was characterized using digital image analysis techniques. Biofilm images were obtained using confocal laser scanning microscopy (CLSM) at various operation points. Structural parameters were then computed from these images using an image analysis software (ISA-2). The structural parameters indicated that the anoxic biofilm was thinner than the aerobic biofilm but the anoxic biofilm was spread out on the membrane surface more uniformly and densely, resulting in the higher membrane fouling. Based on the extracellular polymeric substances (EPS) visualization and quantification, it was also found that EPS, key membrane foulants were spread out more uniformly in the anoxic biofilm in spite of lower amount of EPS compared to that in the aerobic biofilm. PMID- 16360190 TI - Phosphorus removal by an 'active' slag filter-a decade of full scale experience. AB - Active filters, which facilitate phosphorus (P) removal via precipitation and/or adsorption, offer a promising 'appropriate technology' for upgrading small wastewater treatment systems. Research on active filters for P removal using steel slag material has been conducted in laboratories across the world, however, field experiments have been limited and long-term data is practically non existent. This paper presents a decade of experience on P removal by active slag filters at a full-scale treatment plant. During 1993-1994 the filter removed 77% of the total phosphorus (TP), and over the first 5 years of the filter's operation it reduced the mean effluent TP concentration to 2.3 mgl(-1). However during the sixth year of operation P removal was significantly reduced. Over the 11 years of monitoring, 22.4 tonnes of TP was removed by the filter, 19.7 tonnes of this in the first 5-year period. It was determined that the slag material maintained its maximum removal potential until reaching a P-retention ratio of 1.23 kg TP per tonne of slag. This paper provides the first long-term field data for slag filters, and shows that they can provide P removal for a half a decade before filter replacement/rejuvenation is required. PMID- 16360191 TI - Toxicity evaluation of surface water treated with different disinfectants in HepG2 cells. AB - It is well known that water disinfection through chlorination causes the formation of a mixture of disinfection by-products (DBPs), many of which are genotoxic and carcinogenic. To demonstrate the formation of such compounds, a pilot water plant supplied with water from Lake Trasimeno was set up at the waterworks of Castiglione del Lago (PG, Italy). The disinfectants, continuously added to pre-filtered lake water flowing into three different basins, were sodium hypochlorite, chlorine dioxide and peracetic acid, an alternative disinfectant used until now for disinfecting waste waters, but not yet studied for a possible use in drinking water treatment. The aim of this study was to evaluate the formation during the disinfection processes of some toxic compounds that could explain the genotoxic effects of drinking waters. Differently treated waters were concentrated by solid-phase adsorption on silica C(18) columns and toxicity was assessed in a line of human hepatoma cells (HepG2), a metabolically competent cellular line very useful for human risk evaluation. The seasonal variability of the physical-chemical water characteristics (AOX, UV 254 nm, potential formation of THM, pH and temperature) made indispensable experimentation with water samples taken during the various seasons. Autumn waters cause greater toxicity compared to those of other seasons, in particular dilution of the concentrate at 0.5l equivalent of disinfected waters with chlorine dioxide and peracetic acid causes a 55% reduction in cellular vitality while the cellular vitality is over 80% with the all other water concentrates. Moreover it is very interesting underline that non-cytotoxic quantities of the autumnal water concentrates cause, after 2h treatment, a decrease in GSH and a statistically significant increase in oxygen radicals, while after prolonged treatment (24h) cause a GSH increase, without variations in the oxygen radical content. This phenomenon could be interpreted as the cellular adaptation response to an initial oxidative stress. PMID- 16360192 TI - Microbial source tracking using host specific FAME profiles of fecal coliforms. AB - The objective of this study was to investigate the host-specific differences in fatty acid methyl ester (FAME) profiles of fecal coliforms (FC). A known-source library was constructed with 314 FC isolates cultured from 6 possible sources of fecal pollution; 99 isolates from sewage; 29 from bovine; 29 from poultry; 50 from swine; 46 from waterfowl; and 61 from deer. It was found that the hydroxy FAMEs 12:0 2 OH, 12:03 OH, and 14:02 OH were exclusively associated with isolates of human origin. On the other hand, 3 saturated FAMEs, 10:0, 15:0, and 18:0 were found only in isolates from non-human sources, 15:0 being associated with livestock samples only. In addition to the presence of these signature FAMEs, the mean relative masses of 16:1 omega7c and 16:1 ISO/14:03 OH were significantly different between the isolates of human and non-human origins. A linear discriminant function differentiated FC isolates of human origin from those of livestock and wildlife origin at 99% accuracy. These results strongly suggest that the FAME profiles of FC show statistically significant host specificity and may have the potential to be used as a phenotypic microbial source tracking tool. PMID- 16360193 TI - Parallel synthesis of 5-cyano-6-aryl-2-thiouracil derivatives as inhibitors for hepatitis C viral NS5B RNA-dependent RNA polymerase. AB - From random screening of our compound libraries, we identified a hit compound with an IC50 of 27 microM against hepatitis C viral NS5B RNA-dependent RNA polymerase. By using a parallel synthetic strategy, a series of its derivatives were synthesized. From their anti-HCV activity screening, compounds with single digital 3.8 micromolar activity were obtained. PMID- 16360194 TI - Investigation of various physicochemical and environmental parameter influence on pesticide sorption to ditch bed substratum by means of experimental design. AB - Diffuse pollution by pesticide applied in rural catchments may contribute to alter water quality. Besides actions relative to the way the substances are introduced into the environment, it is also possible to limit the contamination by interfering on their transfer pathways from fields to the main river network. Especially, interface areas such as buffer strips or small ditches may play a major part in pesticide diffuse pollution decrease. In ditches a great variety of materials may act as sorbents for organic contaminants: grass, leaves, wood debris or sediments. In this study, laboratory experiments were designed to determine sorption characteristics for three herbicides with different physicochemical properties on sediment and leaves in decay commonly found in agricultural ditches. Sorption capacities were assessed for the herbicides isoproturon, diuron and diflufenican. Experimental design was carried out to investigate the effects of five parameters on herbicide sorption on sediment and dead leaves. These parameters have been chosen according to parallel field experiment needs. Thus, the influence of initial sorbent moisture, herbicide form, i.e. active substance or commercial formulation, water quality (tap or natural ditch water), bromide ions (used as conservative tracers) and solid/liquid ratio have been tested. Within the parameters investigated, pesticide formulation and solid/liquid ratio were the most important parameters affecting pesticide sorption on both ditch materials. PMID- 16360195 TI - The geographic distribution of fluoride in surface and groundwater in Ethiopia with an emphasis on the Rift Valley. AB - This paper analyzes the most extensive database on fluoride distribution in Ethiopia. Of the total 1438 water samples tested, 24.2% had fluoride concentrations above the 1.5 mg/l recommended optimum concentration recommended by WHO. Regionally, by far the highest fluoride levels were recorded in the Rift Valley, where 41.2% of all samples exceeded the 1.5 mg/l level. Only 1.0% of the samples from the central and northwestern highlands and 10.0% in the southeastern highlands exceeded 1.5 mg/l. Larger proportions of deep wells (50.0%) and hot springs (90.0%) than shallow wells (27.2%) and cold springs (12.6%) exceeded the 1.5 mg/l level. The highest fluoride concentrations were recorded for Rift Valley lakes Shala (264.0 mg/l) and Abijata (202.4 mg/l) and the lowest in Lake Tana, and rivers, wells and springs in the highlands. The fluoride concentrations of the Awash River, which originates in the highlands and flows through the Rift Valley, increase downstream, giving concern over the current diversion of high fluoride water from Lake Beseka. Of the various flourosis prevention methods tried in Ethiopia, the treatment of surface water has been shown to be the most feasible and effective for towns and large commercial farms in the Rift Valley, although defluoridation methods should be considered for smaller rural communities. PMID- 16360196 TI - Blood lead level in dogs from urban and rural areas of India and its relation to animal and environmental variables. AB - Lead is a common environmental pollutant with deleterious health effects on human and animal. Industrial and other human activities enhance the lead level in the environment leading to its higher residues in exposed population. The present study was aimed at determining blood lead concentration in dogs from two urban areas and in surrounding rural areas of India and analyzing lead level in dogs in relation to environmental (urban/rural) and animal (age, sex, breed and housing) variables. Blood samples were collected from 305 dogs of either sex from urban (n=277) and unpolluted rural localities (n=28). Irrespective of breed, age and sex, the urban dogs had significantly (P<0.01) higher mean blood lead concentration (0.25+/-0.01 microg/ml) than rural dogs (0.10+/-0.01 microg/ml). The mean blood lead level in stray dogs either from urban or rural locality (0.27+/-0.01 microg/ml) was significantly (P<0.01) higher than that of pets (0.20+/-0.01 microg/ml), and the blood lead concentration was significantly higher in nondescript dogs (0.25+/-0.01 microg/ml) than pedigreed dogs (0.20+/ 0.01 microg/ml). The locality (urban/rural) was the major variable affecting blood lead concentration in dogs. Breed and housing of the dogs of urban areas and only housing (pet/stray) in rural areas significantly (P<0.01) influenced the blood lead concentration in dogs. PMID- 16360197 TI - Age-related differences in heparin response. AB - INTRODUCTION: Major physiological differences in the coagulation system throughout childhood, compared to adults, are well documented. However, the impact of this on anticoagulant drugs is unknown. This study aimed to determine whether heparin therapeutic range determination is affected by the age of plasma donors and whether age-specific therapeutic ranges for heparin therapy may need to be considered in the clinical setting. MATERIALS AND METHODS: Plasma samples were obtained from healthy children and adults, and pooled into age-specific pools. These were spiked with different concentrations of heparin and APTT; Anti Factor Xa and Anti-Factor IIa were measured using standard techniques. The experiments were repeated using three separate plasma pools, and results expressed as means with standard deviations. RESULTS AND CONCLUSIONS: The results show clear age-related differences in APTT for the same Anti-Factor Xa heparin concentration. The differences were more pronounced in younger children, with higher APTT for same Anti-Factor Xa. The Anti-Factor IIa activity of heparin for a given Anti-Factor Xa activity also differed between age-specific plasma pools. This study suggests that when using heparin in children, basic assumptions about the drug mechanism of action and implications for therapeutic ranges need to be considered. PMID- 16360198 TI - HMG-CoA reductase inhibitors may affect thrombin generation by reducing factor VII activity in hyperlipidemic patients. PMID- 16360199 TI - Phase III double-blind randomized trial of radiation therapy for stage IIIb cervical cancer in combination with low- or high-dose Z-100: treatment with immunomodulator, more is not better. AB - BACKGROUND: To evaluate the efficacy of low or high-dose immunomodulator, Z-100, in combination with radiotherapy for cervical cancer. METHODS: Between 1995 and 1999, 221 patients with stage IIIb squamous cell carcinoma of the cervix were randomly assigned to treatment with Z-100 either at 0.2 microg or 40 microg in a double-blind manner in combination with radiotherapy. RESULTS: The 5-year survival of patients with high-dose and low-dose Z-100 was 41.5% (95% CI: 31.7 51.3%) and 58.2% (95% CI: 48.7-67.7%), respectively, showing a 30% reduction in the death rate (hazard ratio: 0.670 [95% CI: 0.458-0.980], P = 0.039). Survival of high-dose group was equivalent to the 4-year survival of the radiotherapy plus hydroxyurea arm (49.7%) of GOG120 study, and that of low-dose group was similar to the survival of the cisplatin-based chemoradiation arm. The progression-free survival was also significantly improved in favor of low-dose group (hazard ratio: 0.667 [95% CI: 0.447-0.997], P = 0.048). The survival of low-dose group was similar to the survival of the cisplatin-based chemoradiation arms of the GOG120 study. CONCLUSIONS: Unexpectedly, the survival of patients with advanced cervical cancer treated by lower dose of Z-100 in combination with radiotherapy was significantly better than those treated with higher dose Z-100, which was equivalent to the survival with radiotherapy alone. The hypothesis that lower dose of Z-100 enhances the efficacy of radiation therapy is now being tested by placebo-controlled randomized trial. PMID- 16360200 TI - Germline genetic polymorphisms of CYP1A1, GSTM1 and GSTT1 genes in Indian cervical cancer: associations with tumor progression, age and human papillomavirus infection. AB - OBJECTIVES: Host genetic factors may play a role in human papillomavirus (HPV) associated tumorigenesis, although the issue continues to be a focus of much debate. Biotransformation is critical in carcinogenic activity of numerous environmental carcinogens. It is therefore possible that polymorphisms of genes producing functional changes in xenobiotic metabolizing enzymes may be susceptible factors in cervical carcinogenesis. This study looked into possible relationships among these factors. METHODS: In this case-control study, we analyzed leukocyte DNA from a total of 312 subjects for germline polymorphisms of CYP1A1 (m1 and m2), GSTM1 and GSTT1 at various stages of the cervical tumor progression spectrum, using PCR and RFLP. RESULTS: Both m1 and m2 polymorphisms of the CYP1A1 gene were more frequent among cases (36.1% for m1 and 38.1% for m2) compared to control subjects (18.2% and 17.6% respectively). The odds ratio of a subject with homozygous CYP1A1 m1 and m2 variant being a case was highest (m1 OR = 4.77 [95% CI = 1.28-17.77]; P = 0.02 and m2 OR = 5.48 [95% CI = 1.49-20.19]; P = 0.011) respectively. The distribution of m1 and m2 CYP1A1 genotypes was also studied as a function of age and in relation to the presence of HPV 16 infection. The risk due to CYP1A1 m1 genotype, when adjusted for HPV status, showed a significantly increased risk (OR = 3.58, 95% CI = 1.88-6.81; P = 0.0001). Similar results were observed in the case of CYP1A1 m2 variant and HPV 16. There was a significant over-representation of both m1 (25.9% vs. 13.9%) and m2 (27.9% vs. 13.3%) polymorphisms in older women (46 years or more). GSTM1 and GSTT1 deletions were also prominent among cases (53.7% and 16.3% respectively) compared to controls (32.7% and 9.7% respectively). A higher proportion of both GSTT1 and GSTM1 deletions were also detected in HPV-16-positive subjects. CONCLUSIONS: These results suggest that polymorphisms in the CYP1A1, GSTM1 and GSTT1 genes may render women more susceptible to the development of cervical cancer. The association between this susceptibility and the presence of human papillomavirus infection further emphasizes the significance of the genetic polymorphisms. PMID- 16360201 TI - The contribution of the hereditary nonpolyposis colorectal cancer syndrome to the development of ovarian cancer. AB - OBJECTIVE: Ovarian cancer has one of the highest fractions of hereditary cases. The hereditary breast and ovarian cancer syndrome, primarily due to mutations in BRCA1 and BRCA2, is the main cause of heredity, but also the hereditary nonpolyposis colorectal cancer (HNPCC) syndrome confers an increased risk of ovarian cancer. In order to clarify the contribution of HNPCC to the development of ovarian cancer, we collected data on family history of cancer and characterized MMR function in a consecutive series of 128 tumors unselected for age at diagnosis and previously characterized for BRCA gene mutations. METHODS: Expression of the MMR proteins MLH1, PMS2, MSH2, and MSH6 was analyzed by immunohistochemistry using tissue microarray sections. Tumors with reduced staining or loss of staining were also analyzed for microsatellite instability (MSI). RESULTS: Loss of MMR protein expression was identified in 3 ovarian cancers, all of which had a MSI-high phenotype. DNA sequence analysis revealed disease-causing germline mutations (deletions of exons 4-6 in MLH1 and a 1 nucleotide deletion in exon 5 of MSH6) in two patients diagnosed at ages 40 and 49 years, both of whom had family histories suggestive of HNPCC. The genetic defect in the third case, which was a 47-year old woman without knowledge about her family history with loss of MLH1/PMS2 expression in the tumor tissue, remains elusive. A family history suggestive of HNPCC was identified in an additional case, but this tumor showed normal, retained MMR protein expression and a microsatellite stable phenotype. CONCLUSIONS: About 2% of ovarian cancer is caused by germline mutations in the MMR-genes, a minor proportion as compared to the contribution of the BRCA-genes (11% in the present series). However, identification of HNPCC patients is important since it allows inclusion of high risk individuals into control programs aimed at preventing the more frequent colorectal and endometrial cancers. Tumors within the HNPCC-spectrum should therefore be included when recording a family history of cancer among patients diagnosed with ovarian cancer. PMID- 16360202 TI - Laparoscopic findings during adnexal surgery in women with a history of nongynecologic malignancy. AB - OBJECTIVES: To describe the results of laparoscopic management of adnexal masses in women with a history of nongynecologic malignancy. METHODS: We conducted a retrospective review of 262 patients with history of prior nongynecologic malignancy who underwent laparoscopy for management of an adnexal mass between 1/1992 and 6/2004. RESULTS: Median patient age at laparoscopy was 55 years (range, 20-91 years), and median BMI was 25 kg/m2 (range, 14-41 kg/m2). Of the 262 patients, 145 (55.3%) had prior abdominal/pelvic surgery. Prior cancer history included breast (202, 77.1%), lymphoma/leukemia (16, 6.1%), colorectal (8, 3.0%), lung (7, 2.7%), multiple myeloma (5, 1.9%), head/neck (5, 1.9%), genitourinary (5, 1.9%), upper gastrointestinal (4, 1.5%), and other (10, 3.8%). Median ovarian mass diameter measured on radiologic imaging was 3.8 cm (range, 0.2-13.5 cm); median CA-125 was 17.0 U/mL (range, 1-7000 U/mL). In all, 49 (18.7%) patients had malignancy identified at laparoscopy, with 30/49 (61.2%) diagnosed with metastatic malignancy to the ovary and 19/49 (38.8%) having a new primary ovarian malignancy. Median tumor diameter and CA-125 were significantly higher in women found to have a malignancy (4.7 vs. 3.7 cm, and 35 vs. 14 U/mL, respectively). Overall, conversion to laparotomy occurred in 34 (12.9%) cases. Twenty-one of 49 (42.9%) patients with malignancy were converted to laparotomy compared with 13/213 (6.1%) when benign disease was noted (P < 0.001). CONCLUSIONS: Approximately 1 in 5 patients with a history of nongynecologic malignancy who were selected for laparoscopic management of an adnexal mass was found to have malignancy, with 60% being metastatic from other primaries. The majority of cases were managed laparoscopically even if malignancy was identified. PMID- 16360203 TI - The effect of cyclosporine on the development and expression of cannabinoid tolerance in mice. AB - Cyclosporine, beside its immunosuppressive action, has several effects on different neuronal functions, such as modulation of neurotransmitter release, the inhibition of nitric oxide synthesis and release, the reduction of cAMP production and inhibition of morphine-induced tolerance. In the present study, the effect of cyclosporine on the expression and development of tolerance to WIN 55,212-2, a cannabinoid receptor agonist, was studied. Intra peritoneal (i.p.) injection of WIN 55,212-2 (2-6 mg/kg) induced time-dependent and dose-dependent analgesia and catalepsy in mice. Administration of cyclosporine (20 mg/kg i.p.), 30 min before WIN 55,212-2 (6 mg/kg i.p.), did not change the analgesic and cataleptic effects of WIN 55,212-2. When WIN 55,212-2 (6 mg/kg i.p.) was injected once a day, animals became completely tolerant to the analgesic and cataleptic effects within five and nine days respectively. Cyclosporine (20 mg/kg i.p.) injected once daily, 30 min before WIN 55,212-2, attenuated the development of tolerance to the analgesic and cataleptic effects of WIN 55,212-2 but did not affect the expression of tolerance. Since cyclosporine given chronically by itself did not alter the analgesia and catalepsy induced by acute administration of WIN 55,212-2, our findings suggest cyclosporine may act with some selectivity on the mechanisms involved in development of cannabinoid tolerance. PMID- 16360204 TI - A facile method to prepare heparin-functionalized nanoparticles for controlled release of growth factors. AB - A new, facile method to prepare the heparin-functionalized PLGA nanoparticle (HEP PLGA NP) for the controlled release of growth factors is developed. This system is composed of PLGA as a hydrophobic core, Pluronic F-127 as a hydrophilic surface layer, and heparin as the functional moiety. HEP-PLGA NPs were prepared by a solvent-diffusion method without chemical modification of the components. The entrapment of heparin molecules was confirmed by a negatively increased zeta potential value and the specific binding affinity to antithrombin III. The average diameter and the surface charge of the nanoparticles were ranged from 139+/-2 to 188+/-4 nm and from -26.0+/-1.1 to -44.1+/-1.3 mV by increasing the amount of heparin during the nanoparticle preparation. Accordingly, the amount of heparin on the nanoparticle increased from 0% to 4.7%. As a model in vitro release experiment, lysozyme was loaded into HEP-PLGA NPs, and a sustained release profile over 2 weeks was obtained with maintaining its bioactivity. The release of rhVEGF, one of the heparin-binding growth factors, showed a more sustained and prolonged profile than that of lysozyme over one month. PMID- 16360205 TI - The use of flow cytometry to discriminate avian lymphocytes from contaminating thrombocytes. AB - Evaluation of peripheral blood mononuclear cells (PBMC) in avian species by flow cytometry is complicated by the presence of large numbers of nucleated thrombocytes. With light scattering properties similar to those of lymphocytes, variations in the proportion of thrombocytes in PBMC can result in apparent shifts in percentages of lymphocyte subpopulations. We have applied a dual labeling procedure for flow cytometric analyses to exclude thrombocytes from the analyzed population by labeling with K55 monoclonal antibody (MAb), which differentially labeled leukocyte and thrombocyte populations. Flow cytometric analyses with K55 labeled chicken PBMC differentiated leukocytes into three populations according to their intensity of fluorescence. Using the Kl MAb, the K55-low population was shown to consist of thrombocytes. Dual-labeling with K55 MAb and MAb specific for B lymphocyte, CD4 or CD8 markers indicated that the K55 intermediate population consisted of lymphocytes. Therefore, concentrations of CD4+ and CD8+ T lymphocytes could be determined from the lymphocyte fraction by gating specifically on the K55 intermediate cells. Selecting cross-reactive chicken MAbs that included K55, this protocol was shown to identify CD4+ and CD8+ T lymphocytes in PBMC of another avian species, the endangered Attwater's prairie chicken. PMID- 16360206 TI - Granulocyte macrophage-colony stimulating factor reduces the affinity of SHP-2 for the ITIM of CLECSF6 in neutrophils: a new mechanism of action for SHP-2. AB - Proteins that bear immunoreceptor tyrosine based inhibitory motifs (ITIM) are believed to participate in the repression of cell activation via phosphatases such as SHP-1, SHP-2 and/or SHIP-1. CLECSF6, also called DCIR, is a transmembrane protein expressed on leukocytes and predominantly on neutrophils that bears one ITIM pattern. This feature confers to CLECSF6 a role in the repression of cell activation. In order to better understand its role in neutrophil signalling, we analysed the binding of phosphatases to the ITIM of CLECSF6. We showed that a peptide bearing the ITIM of CLECSF6 in its phosphorylated form associates with both SHP-1 and SHP-2. Phosphorylated SHP-1 binds the ITIM whereas phosphorylated SHP-2 does not. In addition, granulocyte macrophage-colony stimulating factor (GM CSF) reduces the binding of SHP-2 to the ITIM of CLECSF6 while enhancing the phosphorylation level of SHP-2. GM-CSF is known to recruit SHP-2 to its receptor. These data suggest that the phosphorylation of SHP-2 by GM-CSF promotes the binding of SHP-2 to the GM-CSF receptor to the disadvantage of CLECSF6. Therefore, upon a treatment with GM-CSF, SHP-2 could move from a CLECSF6 associated signalosome with a repressor function to a GM-CSF receptor associated signalosome with an activator function. PMID- 16360207 TI - Long-term outcomes of photorefractive keratectomy for anisometropic amblyopia in children. AB - PURPOSE: To evaluate the long-term visual acuity (VA) and refractive error responses to excimer laser photorefractive keratectomy (PRK) for treatment of anisometropic amblyopia in children. DESIGN: Prospective interventional case control study. PARTICIPANTS: Eleven children, 2 to 11 years old, with anisometropic amblyopia who were noncompliant with conventional therapy with glasses or contact lenses and occlusion therapy were treated with PRK. A cohort derived retrospectively of 13 compliant and 10 noncompliant children with refractive errors similar to those of the PRK group who were treated with traditional anisometropic amblyopia therapy served as control groups. INTERVENTION: Photorefractive keratectomy for the eye with the higher refractive error. MAIN OUTCOME MEASURES: (1) Refractive error reduction and stability in the treated eye, (2) cycloplegic refraction, (3) VA, (4) stereoacuity, and (5) corneal haze up to 3 years after PRK. Compliant and noncompliant children with anisometropia amblyopia were analyzed as controls for refractive error and VA. RESULTS: Preoperative refractive errors were -13.70 diopters (D) (+/-3.77) for the myopic group and +4.75 D (+/-0.50) for the hyperopic group. Mean postoperative refractive errors at last follow-up (mean, 31 months) were -3.55 D (+/-2.2.5) and +1.41 D (+/-1.07) for the myopic and hyperopic groups, respectively. At last follow-up, cycloplegic refractions in 4 (50%) of 8 myopes and all hyperopes (100%) were within 3 D of that of the fellow eye. Five (63%) of 8 myopic children achieved a refraction within 2 D of the target refraction. Two (67%) of 3 hyperopic patients maintained their refractions within 2 D of the target. Refractive regressions (from 1 year after surgery to last follow-up) were 0.50+/-1.41 D (myopes) and 0.60+/-0.57 D (hyperopes). Seven children (77%) were able to perform psychophysical VA testing preoperatively and postoperatively. Five (71%) of the 7 children had uncorrected VA improvement of at least 2 lines, and 4 (57%) of 7 had best spectacle-corrected VA improvement of at least 2 lines, with 1 improving 7 lines. Five (55%) of 9 children had improvement of their stereoacuity at last follow-up. Subepithelial corneal haze remained negligible. The mean final VA of the PRK group was significantly better than that of the noncompliant control group (P = 0.003). The mean final refractive error for both myopic and hyperopic groups was also significantly better that that of the control groups (P = 0.007 and P<0.0001, respectively). CONCLUSIONS: Photorefractive keratectomy for severe anisometropic amblyopia in children resulted in long-term stable reduction in refractive error and improvement in VA and stereopsis, with negligible persistent corneal haze. PMID- 16360208 TI - Attachment of Acanthamoeba to first- and second-generation silicone hydrogel contact lenses. AB - PURPOSE: To investigate the attachment of Acanthamoeba to first- and second generation silicone hydrogel contact lenses, and to determine if patient wear or the presence of a bacterial biofilm coating affects attachment characteristics. DESIGN: Experimental study. PARTICIPANTS AND CONTROLS: Attachment to the silicone hydrogel lenses was compared with that to a conventional hydrogel control lens. Sixteen replicates (n = 16) were carried out for unworn, worn, and biofilm-coated lenses of each type. METHODS: Unworn, worn, and Pseudomonas aeruginosa biofilm coated first-generation (lotrafilcon A) and second-generation (galyfilcon A) silicone hydrogel and conventional hydrogel (etafilcon A) lens quarters were incubated for 90 minutes in suspensions of plate-cultured Acanthamoeba castellanii trophozoites. MAIN OUTCOME MEASURES: Trophozoites attached to one surface of each lens quarter were counted by direct light microscopy. Logarithmic transformation of data allowed the use of parametric analysis of variance for statistical analysis. RESULTS: Attachment of Acanthamoeba was affected significantly by lens material type (P<0.001), with higher numbers of trophozoites attaching to the first-generation lotrafilcon A silicone hydrogel lens, compared with the second-generation galyfilcon A lens and the conventional hydrogel lens. Attachments to the latter 2 lenses did not differ significantly from each other (P = 0.126). Patient wear and the presence of a bacterial biofilm had no significant effect on attachment to the lotrafilcon A lens (P = 0.426) but did significantly increase attachment to the galyfilcon A (P<0.001) and the etafilcon A (P = 0.009) lenses; attachment to the latter 2 lenses was still significantly less than that found with the first-generation silicone hydrogel (P<0.001). CONCLUSIONS: Acanthamoeba demonstrated a significantly greater affinity for the first-generation silicone hydrogel lens as compared with the second-generation silicone hydrogel and the conventional hydrogel. If exposed to Acanthamoeba (e.g., when showering or swimming, through noncontinuous wear and ineffective lens care regimes), first-generation silicone hydrogel lenses may promote a greater risk of Acanthamoeba infection due to the enhanced attachment characteristics of this lens material. However, prospective studies in patients are required to determine if these experimental results are clinically significant. PMID- 16360209 TI - Verteporfin therapy combined with intravitreal triamcinolone in all types of choroidal neovascularization due to age-related macular degeneration. AB - OBJECTIVE: To evaluate the efficacy and safety of photodynamic therapy with verteporfin combined with intravitreal triamcinolone in choroidal neovascularization secondary to age-related macular degeneration (AMD). DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: One hundred eighty-four patients undergoing treatment for neovascular AMD at one retinal referral center. METHODS: One hundred eighty-four eyes of 184 consecutive patients (63.6% female, 36.4% male) with a mean age of 76.5 years and a follow-up of a median of 38.8 weeks (range, 12-103) were included in a case series. One hundred forty-eight (80.4%) patients had subfoveal choroidal neovascularization, 19 patients (10.3%) had juxtafoveal choroidal neovascularization, and 17 patients (9.2%) had extrafoveal choroidal neovascularization. Verteporfin photodynamic therapy was performed using the recommended standard procedure. A solution containing 25 mg of triamcinolone was injected intravitreally 16 hours after photodynamic therapy in 184 patients. The combined therapy procedure was repeated at the 3-month follow-up visits whenever persistent choroidal neovascularization leakage was documented angiographically. MAIN OUTCOME MEASURES: Mean change in best-refracted visual acuity (VA) between baseline and the last visit, and number of treatments necessary to achieve absence of leakage. RESULTS: Visual acuity improved in the majority of patients (baseline VA, mean 20/125) by a mean increase of 1.22 Snellen lines and 1.43 lines using laser interferometry (P<0.01). The mean number of required treatments was 1.21. Twenty-three eyes (12.5%) required 2 treatments, 6 eyes (3.26%) required 3 treatments, and 1 eye (0.5%) required 4 treatments. The combination treatment including laser and intravitreal steroid administration was well tolerated. Forty-six patients (25%) required glaucoma therapy due to a transient steroid-induced intraocular pressure (IOP) increase. Twelve patients (6.5%) were on topical medication for preexisting glaucoma. Two patients (1%) whose IOP increase could not be controlled with topical therapy required surgery. CONCLUSIONS: Verteporfin photodynamic therapy combined with intravitreal triamcinolone may improve the outcome of standard verteporfin photodynamic therapy in the treatment of choroidal neovascularization secondary to AMD. A significant improvement in VA was observed in a majority of treated patients and was maintained during the maximum follow-up. In addition, retreatment rates were lower than anticipated. PMID- 16360210 TI - Microbial keratitis predisposing factors and morbidity. AB - PURPOSE: To examine predisposing factors, treatment costs, and visual outcome of microbial keratitis in an ophthalmic casualty and inpatient population. DESIGN: Retrospective medical records review. PARTICIPANTS: Fifteen- to 64-year-olds with microbial keratitis treated at the Royal Victorian Eye and Ear Hospital between May 2001 and April 2003 (n = 291). METHODS: Risk factors were identified from patient files. Demographic, clinical, and microbiological data; severity; outpatient visits; hospital bed days; and vision loss were examined. MAIN OUTCOME MEASURES: Cost to treat (Australian dollars), vision loss, and factors influencing these outcomes. RESULTS: Ocular trauma (106/291 [36.4%]) and contact lens (CL) wear (98/291 [33.7%]) were the most commonly identified predisposing factors; 18 (6.1%) had multiple predisposing factors; 17 (5.8%), ocular surface disease; 20 (6.9%), herpetic eye disease; 4 (1.4%), systemic associations; 5 (1.7%), other; and 23 (7.9%), unknown cause. Of trauma cases, 90.6% involved males, compared with 44% to 57% for other groups (P<0.001). Contact lens wearers were younger than the other groups--mean age 30 years, compared with 40 to 47 years (P<0.01). Gram-negative organisms were isolated more frequently in CL wearers than trauma cases (18.7% vs. 6.5%, P = 0.01). The number of outpatient visits was 4+/-1 (median +/- interquartile range), and 19.6% (57/291) were hospitalized for 5+/-2 days. Hospital resource use and vision loss were similar for predisposing factors but differed by causative microorganism. Eighty-eight percent of cases were scraped: acanthamoeba keratitis was the most expensive to treat, followed by fungal and herpetic keratitis and, lastly, culture-proven bacterial keratitis or culture-negative cases (P<0.0001). After treatment, 21.7% exhibited >2 lines of vision loss, and 1.6% of cases had > or =10 lines of vision loss. Vision loss was associated with clinical severity (P = 0.005). CONCLUSIONS: Ocular trauma and CL wear are the major predisposing factors for microbial keratitis in this age range. These cases require significant hospital resources during treatment, and the keratitis may result in loss of vision. PMID- 16360211 TI - Topical 0.1% prednisolone lowers nerve growth factor expression in keratoconjunctivitis sicca patients. AB - PURPOSE: To compare nerve growth factor (NGF) levels in tears and on the ocular surfaces of normal control and non-Sjogren's type keratoconjunctivitis sicca subjects, and to investigate the effect of 0.1% prednisolone eyedrops on NGF levels in keratoconjunctivitis sicca patients. DESIGN: Prospective, double masked, randomized, comparative clinical trial. PARTICIPANTS: Forty-one keratoconjunctivitis sicca patients and 23 age- and gender-matched healthy subjects. METHODS: Baseline tear NGF levels were measured in keratoconjunctivitis sicca patients and healthy control subjects using enzyme-linked immunosorbent assays. Keratoconjunctivitis sicca patients received 0.1% prednisolone drops in one eye and 0.1% hyaluronic acid drops in the other, 3 times a day for 28 days. Also, impression cytology (IC) and immunostaining for NGF on conjunctival epithelium were performed on both groups. MAIN OUTCOME MEASURES: Tear NGF/total tear protein (TP) concentration ratio, IC and NGF immunocytologic staining, subjective symptom scale, tear breakup time, and Schirmer values. RESULTS: Keratoconjunctivitis sicca patients were found to have baseline tear NGF concentrations higher than those of age- and gender-matched healthy control subjects (65.9+/-14.5 vs. 122.1+/-45.3 pg/mug, P<0.0001). In keratoconjunctivitis sicca patients, prednisolone treatment for 28 days resulted in a decrease in tear NGF levels, symptom scores, and IC scores, whereas hyaluronic acid treatment had no such effect (68.2+/-25.0 pg/mug vs. 108.0+/-43.4 pg/mug, P<0.0001 for tear NGF/TP ratio; 2.16+/-1.01 vs. 3.39+/-1.50, P = 0.0014 for symptom scale; 1.05+/ 0.67 vs. 1.61+/-0.86, P = 0.0317 for IC). Measurements taken at both 14 and 28 days indicated that neither prednisolone nor hyaluronic acid treatment affected breakup time or Schirmer values. CONCLUSION: Keratoconjunctivitis sicca patients showed elevated levels of tear NGF, which were decreased by treatment with 0.1% prednisolone. These data suggest that ocular surface NGF may play an important role in ocular surface inflammation processes associated with dry eyes. PMID- 16360212 TI - Natural killer/T-cell lymphoma with ocular and adnexal involvement. AB - PURPOSE: To review the clinical, radiological, and histopathologic features in 8 patients with natural killer/T-cell lymphoma (NKTL) involving the orbit and/or ocular adnexa, and to describe the responses of these patients to various treatment regimens. DESIGN: Retrospective observational case series. PARTICIPANTS: Eight patients (5 male, 3 female) with NKTL involving the orbit and/or ocular adnexa were identified from 1999 through 2005. The mean age at presentation was 45 years (range, 26-65). METHODS: We retrospectively identified patients with NKTL of the ocular adnexa treated in the authors' medical centers from 1999 through 2004 using computerized diagnostic index retrieval. The clinical records and radiologic studies were analyzed to define modes of presentation and progression, response to therapy, and areas of anatomic involvement. Histopathologic findings, including the presence of CD3, CD56, and Epstein-Barr virus-encoded mRNA in each patient, were reviewed. MAIN OUTCOME MEASUREMENTS: Time of survival from presentation to last known follow-up and tumor-related death. RESULTS: Four of the 8 patients (50%) with NKTL involving the orbit or ocular adnexa had systemic involvement at presentation. Five of the 8 patients (62.5%) had concurrent sinonasal involvement, whereas 3 (37.5%) had orbital involvement alone. All lesions demonstrated CD3, CD56, and/or Epstein Barr virus positivity on immunopathology studies. Therapy consisted of various chemotherapeutic regimens typically employed in the treatment of non-Hodgkins lymphoma, steroids, surgical intervention, and radiation. Seven (87.5%) patients died 5 weeks to 13 months after presentation, and 1 (12.5%) is alive without disease (5-year follow-up). CONCLUSIONS: Natural killer/T-cell orbital lymphoma is a rare Epstein-Barr virus-associated neoplasm that may occur with or without associated sinonasal involvement. Our series, the largest cohort reported to date, demonstrates the high lethality of this condition despite aggressive conventional therapy, suggesting that new treatment options should be considered early in the course of treatment of patients with this disorder. PMID- 16360213 TI - Incidence of intraocular pressure spike and other adverse events after vitreoretinal surgery. AB - PURPOSE: To determine the incidence of significant postoperative intraocular pressure (IOP) elevation and other adverse medical events during the first postoperative day in patients undergoing inpatient vitreoretinal surgery. DESIGN: Noncomparative interventional case series. PARTICIPANTS: Two hundred two consecutive patients were admitted prospectively as inpatients after vitreoretinal surgery (vitrectomy and/or scleral buckling). METHODS: Intraocular pressure was measured preoperatively, 5 to 12 hours postoperatively, and on postoperative day 1. Additionally, pain, nausea, and serious systemic adverse events were monitored. RESULTS: Seventeen patients (8.4%) had a 5- to 12-hour IOP measurement above 29 mmHg. Thirty patients (14.8%) had a postoperative day 1 IOP measurement above 29 mmHg. Three patients (1.5%) required a parenteral narcotic to achieve adequate analgesia. Nine patients (4.5%) utilized a parenteral antiemetic for nausea control. Six patients (3.0%) had a hyperglycemic event. No serious systemic adverse events occurred in the early postoperative period. CONCLUSIONS: The incidence of a significant IOP spike or systemic adverse events in the early postoperative period in patients undergoing vitreoretinal surgery is low. PMID- 16360214 TI - Efficacy of amphotericin B or amphotericin B-intralipid in combination with caspofungin against experimental aspergillosis. AB - OBJECTIVE: Infections caused by Aspergillus species are increasing in importance, especially among immunocompromised hosts. The objective of this study was to evaluate the efficacy of combination treatment consisting of the polyene amphotericin B (AMB) or amphotericin B-intralipid admixture (AMB-IL) and the echinocandin caspofungin (CAS) in experimental murine systemic aspergillosis. Inhibition of synthesis of a major component of the fungal cell wall and an effect on the cell membrane, by combining echinocandin and a polyene, may result in a synergistic interaction in vitro and in vivo against Aspergillus fumigatus. METHODS: ICR mice were immunosuppressed by intraperitoneal (ip) administration of cyclophosphamide (CY). Three days post-CY administration the mice were inoculated intravenously (iv) with A. fumigatus conidia. Infection and treatment were evaluated during an observation period of 30 days in terms of mortality (survival rate and mean survival time) and morbidity (quantitative determination of fungal burden, histopathology, and detection of serum galactomannan). RESULTS: The data showed that combined CAS and AMB or AMB-IL treatment increased the survival of the mice (up to 69.2%) as compared to those treated with each agent alone (44.4, 40.7 and 50%, respectively), and prolonged their mean survival time to 22.5 days. These combinations also resulted in reduction of fungal burden in organs, and decrease in serum galactomannan. CONCLUSION: The successful results obtained in the experimental animal model of this study may possibly open the way to more effective management of aspergillosis in humans. PMID- 16360215 TI - Streptococcal toxic shock syndrome after a dental implant treatment: should patients receive antibiotic prophylaxis? PMID- 16360216 TI - An assessment of patient preference and adherence to treatment with Wellbutrin SR: a web-based survey. AB - BACKGROUND: Research has shown that lack of treatment adherence is a serious problem, especially among patients with psychiatric disorders. The current study was conducted to assess adherence and patient preference among individuals taking Wellbutrin SR (bupropion) for depression, as well as their interest in a once daily formulation of bupropion. METHODS: A 20-item web-based survey was administered to 527 individuals (276 men and 251 women) recruited through an online panel. All participants were at least 18 years of age, diagnosed with major depressive disorder, and had been taking Wellbutrin SR for at least 6 weeks. Survey items addressed treatment regimen, adherence, satisfaction with Wellbutrin SR, and interest in a once-daily formulation of bupropion. RESULTS: The majority of respondents reported taking Wellbutrin SR twice a day (67%). Only 15% of once-daily users were nonadherent compared to 37% of twice-daily users and 65% of thrice-daily users. The most common reason reported for missing a dose of Wellbutrin SR was simply forgetting to take it (49% of twice-daily users and 65% of thrice-daily users). Results indicated that 77% of twice-daily users and 94% of thrice-daily users were interested in a once-daily formula. CONCLUSIONS: A reduction in dosing frequency is favored by Wellbutrin SR users and likely to improve their adherence to treatment. Because greater adherence has been shown to facilitate symptom relief, improvements in quality of life, and reductions in healthcare expenses, the results of this study support the value of the recently released once-daily formulation, Wellbutrin XL. PMID- 16360217 TI - Is there evidence for social rhythm instability in people at risk for affective disorders? AB - Social rhythm disruptions are thought to be related to the etiology of affective symptoms. 'Hypomanic personality' and 'rigidity' are hypothesized to be risk factors for affective disorders. We examined whether people scoring high on such scales would demonstrate instability of social rhythms and sleep. In a short-term prospective diary study with one group factor, the following three groups were selected from a non-university student sample: 'bipolar risk' (scoring high on the 'Hypomanic Personality Scale'; n=56); 'Unipolar risk' (scoring high on the 'Rigidity Scale'; n=37); and a control group (scoring low on both scales; n=48). The participants completed ratings of their activities and sleep for 28 days. People at risk for bipolar disorders showed a lower regularity of daily activities than controls. Their sleeping pattern was not characterized by fewer but by more variable hours of sleep. The unipolar risk group did not differ from the control group at all. Despite some limitations, there is partial evidence for social rhythm and sleep irregularities in people putatively at risk for bipolar disorders. Further research is, however, needed to replicate and extend these results. PMID- 16360218 TI - Elevated cytokine levels in children with autism spectrum disorder. AB - This study compared production of IL-2, IFN-gamma, IL-4, IL-13, IL-5 and IL-10 in peripheral blood mononuclear cells from 20 children with autism spectrum disorder to those from matched controls. Levels of all Th2 cytokines were significantly higher in cases after incubation in media alone, but the IFN-gamma/IL-13 ratio was not significantly different between cases and controls. Cases had significantly higher IL-13/IL-10 and IFN-gamma/IL-10 than controls. CONCLUSION: Children with ASD had increased activation of both Th2 and Th1 arms of the adaptive immune response, with a Th2 predominance, and without the compensatory increase in the regulatory cytokine IL-10. PMID- 16360219 TI - Detection of HCV salivary antibodies by a simple and rapid test. AB - Hepatitis C (HCV) is common in developing countries, where blood sampling and expensive sophisticated methods for detection are less available. Hemodialysis patients have high prevalence of HCV and may resemble sick populations in developing countries in relation to immunosuppression and antibodies production. For these reasons anti-HCV antibodies were assayed in saliva of hemodialysis patients by ImmunoComb II assay that is less laborious, relatively inexpensive and easy to perform If the findings are confirmed by larger studies this method may be useful especially in developing countries. Serum and saliva samples were obtained from 37 hemodialysis patients and assayed by ImmunoComb II kit. In positive PCR patients the saliva test had 100% sensitivity, which was as good as serum anti-HCV Axsym testing. Saliva testing had a similar or better specificity than the serum method. PMID- 16360220 TI - A cost-effective algorithm for the diagnosis of Hepatitis C virus infection and prediction of HCV viremia in Cameroon. AB - Conventional tests for antibody to Hepatitis C virus (HCV) and HCV RNA require considerable time before results are available, remain very expensive and are not adapted to many sub-Saharan African countries where HCV is endemic. The aim of this study was to evaluate the accuracy of an algorithm consisting of two HCV rapid tests to diagnose and predict HCV viremia in patients in Cameroon. Three hundred and twenty nine plasma samples were screened by two HCV rapid tests (ImmunoComb II HCV, PBS Orgenics and Hexagon HCV, Human). Previous evaluation of these samples for HCV antibodies (anti-HCV) by conventional third generation ELISA, considered as a reference test, indicated that 168 were anti-HCV negative and 161 positive. Among the 161 anti-HCV positive plasma, 114 (71%) were HCV RNA positive by RT-PCR assay. The ImmunoComb II HCV test provided the more sensitive detection of anti-HCV (sensitivity: 99.4% with a 95% CI = 96-100%). Surprisingly, the second HCV rapid test, Hexagon HCV, showed a high capacity to identify non viremic subjects amongst anti-HCV positive cases (93.6% [95% CI: 82-99%]). These results suggest an algorithm using ImmunoComb II HCV as a first test to screen anti-HCV positive subjects, and Hexagon HCV as a second test to discriminate between viremic and non-viremic HCV seropositive subjects. PMID- 16360221 TI - Inhibition of neprilysin by thiorphan (i.c.v.) causes an accumulation of amyloid beta and impairment of learning and memory. AB - An accumulation of amyloid beta peptide (Abeta) due to an imbalance between anabolism and catabolism triggers Alzheimer's disease (AD). Neprilysin is a rate limiting peptidase, which participates in the catabolism of Abeta in brain. We investigated whether rats continuously infused with thiorphan, a specific inhibitor for neprilysin, into the cerebral ventricle cause cognitive dysfunction, with an accumulation of Abeta in the brain. Thiorphan-infused rats displayed significant cognitive dysfunction in the ability to discriminate in the object recognition test and spatial memory in the water maze test, but not in other hippocampus-dependent learning and memory tasks. Thiorphan infusion also elevated the Abeta40 level in the insoluble fraction of the cerebral cortex, but not that of the hippocampus. There was no significant difference in the nicotine stimulated release of acetylcholine in the hippocampus between vehicle- and thiorphan-infused rats. These results indicate that continuous infusion of thiorphan into the cerebral ventricle causes cognitive dysfunction by raising the level of Abeta in the cerebral cortex, and suggest that a reduction of neprilysin activity contribute to the deposition of Abeta and development of AD. PMID- 16360222 TI - 5,7-DHT lesion of the dorsal raphe nuclei impairs object recognition but not affective behavior and corticosterone response to stressor in the rat. AB - Previous studies with acute tryptophan depletion, leading to transient central 5 HT reductions, showed no effects on affective behavior but impaired object memory. In the present study, the behavioral effects of a 5,7-dihydroxytryptamine (5,7-DHT) lesion in the dorsal raphe were evaluated in animal models of anxiety (open field test), depression (forced swimming test), behavioral inhibition (discrete fixed interval test) and cognition (object recognition task). The corticosterone response to a stress condition was examined at several intervals after 5,7-DHT treatment. The substantial reduction in neuronal 5-HT markers in the dorsal raphe did not affect anxiety-related, depressive-like or impulsive behavior. Compared to the SHAM group, the lesioned rats showed a lower response latency to obtain a reward, indicating a quick and accurate reaction to a stimulus. No differences were found in the progressive ratio test for food motivation. A marked impairment in object recognition was found. The 5,7-DHT treatment did not affect the corticosterone response to a stressful situation. Overall, these results corroborate studies with acute tryptophan depletion suggesting a role of 5-HT in object memory, but not affective behavior. PMID- 16360223 TI - Myocardial tissue Doppler and long axis function in the fetal heart. AB - Current echocardiographic assessment of fetal ventricular function uses relatively crude measures and this has led to inconsistencies in assessment of diastolic function. Long axis parameters may be more insightful. OBJECTIVE: To describe fetal long axis cardiac function and construct reference ranges. DESIGN: 159 normal fetuses were studied cross-sectionally using long axis M-mode to measure displacement of the atrioventricular ring and pulsed wave Doppler ultrasound to record myocardial tissue Doppler velocities at the base of the heart. RESULTS: There was a strong, almost linear gestational relationship for all but late left basal myocardial velocities. Systolic amplitude of the atrioventricular ring increased significantly. Wall thickness increased with gestation and heart rate reduced. Late diastolic mitral and tricuspid flow velocities showed no age-dependence. Right ventricular filling (p<0.0001) and myocardial lengthening (p<0.0001) and shortening velocities (p<0.0001) were higher than left. Aortic velocities were higher than pulmonary (p<0.0001). Right ventricular long axis amplitude was greater than left (p<0.0001). There were no significant differences in right (RV) and left (LV) ventricular and septal thicknesses (RV 2.5 mm [1.2-4.1]; LV (2.4 mm [1.0-4.0] and septum 2.6 mm [1.0 4.3]). Left and right myocardial tissue Doppler velocities showed similar maturation in systole (LV 0.14 vs. RV 0.10 cm/s/week) and early diastole (LV 0.16 vs. RV 0.14 cm/s/week). CONCLUSIONS: This study demonstrates maturational changes occur in both fetal systolic and diastolic ventricular function and moreover the maturational rate of myocardial tissue velocities is similar in both ventricles, despite differential loading conditions. PMID- 16360224 TI - Atrial fibrillation: the Chinese experience. PMID- 16360225 TI - Direct and indirect comparison meta-analysis demonstrates the superiority of sirolimus- versus paclitaxel-eluting stents across 5854 patients. AB - There is ongoing debate to identify the most effective, safe and cost-beneficial drug-eluting stent, between the two currently approved and used devices, i.e. sirolimus-eluting stents (SES) and paclitaxel-eluting stents (PES). To date, head to-head comparison studies of SES vs PES have been however limited by relatively small sample sizes and the low number of events typically associated with these highly effective coronary devices. To overcome the drawbacks of single trials, direct and indirect comparison meta-analyses have been designed and conducted to thoroughly compare sirolimus- vs paclitaxel eluting-stents. This article provides results of a pooled analysis of such indirect and direct comparisons, definitively demonstrating across 5854 patients the superiority of SES in comparison to PES (odds ratio 0.62 [95% confidence interval 0.50-0.75], p<0.0001 for binary angiographic restenosis, and odds ratio 0.66 [0.52-0.84], p=0.0008 for target lesion revascularization). Indeed, such combination of direct and indirect comparisons should also be envisaged to soundly and timely appraise the next generation of drug-eluting stents. PMID- 16360226 TI - Evaluation of underdosage in the external photon beam radiotherapy of glottic carcinoma: Monte Carlo study. AB - PURPOSE: Underdosage in the human larynx may be the true factor behind the decrease in local control rates. PATIENTS AND METHODS: To evaluate underdosage with Monte Carlo a CT-based geometrical model of the patient's neck (mathematical neck) was created. Dose was calculated for a pair of 6 Me V parallel-opposed photon beams modulated with 15 degree steel wedges. RESULTS: At least 5% of volume of 3.5 cm(3) hypothetical tumor near the air wall of the larynx receives less than 86% of the maximum tumor dose. The same volume received less than 91% of the maximum tumor dose when the mathematical neck had no air cavities. CONCLUSIONS: We conclude the significant underdosage at the air-tissue interface in the larynx occurs in traditional radiotherapy treatments, especially in the glottic part of the larynx. PMID- 16360227 TI - Blunted hypothalamic-pituitary-adrenocortical axis responsivity to stress in persons with a family history of alcoholism. AB - Abstinent alcoholics show a blunted stress cortisol response that may be a consequence of drinking or a preexisting risk marker. We tested cortisol responses to psychological stress in 186 18-30 year-old, healthy social drinkers having no personal history of alcohol or drug dependence, 91 of whom had one or two alcoholic parents (FH+) and 95 having no family alcoholism for two generations (FH-). We predicted that, similar to alcoholic patients, the FH+ would have reduced stress cortisol responses that would be partially determined by their temperament characteristics, specifically antisocial tendencies as measured by the California Psychological Inventory. On a stress day, subjects performed continuous simulated public speaking and mental arithmetic tasks for 45 min, and on a control day they sat and rested for the same time period. The FH+ who were low in sociability had smaller cortisol responses than FH-, high sociability persons (t=2.27, p=.02). These two groups were not different in diurnal cortisol secretion patterns or affective responses to the stressors. Persons with a familial risk for alcoholism who have more antisocial tendencies may have altered central nervous system responses to emotionally relevant social challenges. Disrupted cortisol stress responses may serve as a risk marker for the development of substance use disorders. PMID- 16360228 TI - Genetic integrity of somaclonal variants in tea (Camellia sinensis (L.) O Kuntze) as revealed by inter simple sequence repeats. AB - Adoption of inter simple sequence repeats (ISSR) technique to analyze the genetic variability of somatic embryo derived tea plants was evaluated. Morphological characterisation of the field grown plants revealed no identical character aligning with the parent, UPASI-10. Out of 40 primers, 15 exhibited concurrent polymorphism were selected for the study. Genetic variability of somaclones derived from single line cotyledonary culture ranged from 33.0 to 55.0%. A unique fragment of 1.2Kb was visible in majority of the accessions whereas the fragments below the length of 0.6Kb were noticed only in 50% of the variants. Out of 120 interactions attempted using Pearson's coefficient correlation, only 9.2% of somaclones exhibited significant similarity at genetic level. Dendrogram constructed based on simple matching coefficient revealed a distance of 2.257 3.317 between the final clusters. This strengthens the existence of wide genetic variation among the somaclones. PMID- 16360229 TI - Non-linear pharmacodynamics in a non-viral gene delivery system: positive non linear relationship between dose and transfection efficiency. AB - A remarkable non-linearity was found between dose and transfection activities of non-viral gene delivery systems, such as a Lipofectamine/DNA complex and an octaarginine-modified multifunctional envelope-type nano device (R8-MEND). We measured the nuclear delivery of pDNA to distinguish the non-linearity in intracellular pharmacokinetics or pharmacodynamics after transfection with R8 MEND at different doses. A remarkable positive non-linearity was found in the pharmacodynamics when the dose was increased. Even dummy pDNA enhanced the efficiency of transcription and/or translation per pDNA in the nucleus, but empty liposomes did not. These results suggest the importance of controlled pharmacodynamics as well as the importance of intracellular pharmacokinetics for the rational design of non-viral gene delivery systems. PMID- 16360230 TI - Overlap syndrome of primary biliary cirrhosis and autoimmune hepatitis: a foray across diagnostic boundaries. PMID- 16360231 TI - Antiviral hepatitis and antiretroviral drug interactions. AB - More and more HIV-infected patients are treated for viral hepatitis, increasing interactions. HEPATITIS C: The concomitant use of didanosine and ribavirin increases the risk of mitochondrial toxicity, responsible for pancreatitis and/or lactic acidosis. Lactic acidosis is characterized by a high mortality rate. Thus, didanosine, but also stavudine, should not be co-administered with ribavirin. Cases of hepatic decompensation have been reported in cirrhotics concomitantly receiving ribavirin and didanosine. Thus, this co-admininistration should be contraindicated in patients with advanced liver fibrosis. Anemia is a frequent side effect of ribavirin. In patients with zidovudine-related anemia, this drug should be discontinued before prescribing ribavirin. Erythropoietin may help to improve the haemoglobin level. HEPATITIS B: Adefovir significantly decreases the plasma levels of saquinavir. Pancreatitis may occur with the co-administration of didanosine and tenofovir. Thus this co-administration should be avoided. Atazanavir concentrations are decreased when tenofovir is co-administered. Thus, atazanavir should be boosted with ritonavir, when combined with tenofovir. Atazanavir increases the concentrations of tenofovir, with the potential risk of increasing the adverse events of tenofovir, including renal disorders. Tenofovir area under the curve is increased if lopinavir-ritonavir are co-administered. The main interactions, with a fatal risk, are observed with didanosine, when co administered with ribavirin (hepatitis C) or with tenofovir (hepatitis B). Anemia is frequent, but usually moderate, when zidovudine is co-administered with ribavirin. Other interactions are usually easy to manage. PMID- 16360232 TI - Treatment algorithm for the management of hepatitis C in HIV-coinfected persons. AB - In the era of highly effective antiretroviral therapy (ART), HCV-related liver disease has emerged as a significant cause of morbidity and mortality. Accordingly, expert panels have recommend that coinfected patients undergo medical evaluation for HCV-related liver disease, consideration for HCV treatment and, if indicated, orthotopic liver transplantation. While the treatment of such patients is complicated by medical, and psychiatric comorbidities, HIV disease, and concurrent antiretroviral therapy, randomized controlled trials support the safety, tolerability and efficacy of HCV treatment with peginterferon alfa (PEG IFN) plus ribavirin (RBV) in HIV-infected persons. Although, the available data has led to consensus among experts regarding the need to medically manage HCV disease in HIV-infected persons, uncertainty remains regarding the best treatment algorithm for coinfected patients. PMID- 16360233 TI - Production of infectious hepatitis C virus in tissue culture: a breakthrough for basic and applied research. PMID- 16360234 TI - Different expression patterns of mucin core proteins and cytokeratins during intrahepatic cholangiocarcinogenesis from biliary intraepithelial neoplasia and intraductal papillary neoplasm of the bile duct--an immunohistochemical study of 110 cases of hepatolithiasis. AB - BACKGROUND/AIMS: Two types of neoplastic lesions preceding invasive intrahepatic cholangiocarcinoma (ICC) are identified: a flat-type neoplastic lesion called 'biliary intraepithelial neoplasia (BilIN)' and an intraductal papillary neoplasm of the bile duct (IPN-B). Multi-step carcinogenesis has been suggested in both lesions, although phenotypic changes during this process remain unclarified. METHODS: We immunohistochemically examined expression patterns of MUC1, MUC2, MUC5AC, cytokeratin 7 (CK7), and CK20 in BilIN, IPN-B, and ICC in 110 cases of hepatolithiasis. RESULTS: Thirty-seven cases of ICC in hepatolithiasis were divided into 18 tubular adenocarcinomas with BilIN, 10 tubular adenocarcinomas with IPN-B and nine colloid carcinomas with IPN-B. Carcinogenesis via BilIN was characterized by MUC2-/CK7+/CK20-with increasing MUC1 expression. IPN-B was characterized by the intestinal phenotype (MUC2+/CK20+), and carcinogenesis leading to tubular adenocarcinoma was associated with increasing MUC1 expression and that to colloid carcinoma with MUC1-negativity. Pathological stages of tubular adenocarcinoma of ICC with BilIN or IPN-B were more advanced than those of colloid carcinoma with IPN-B. CONCLUSIONS: Immunophenotypes of MUCs and cytokeratins might characterize three cholangiocarcinogenetic pathways in hepatolithiasis. Increased expression of MUC1 in BilIN and also IPN-B is associated with tubular adenocarcinoma, while colloid carcinoma in IPN-B is characterized by MUC1-negativity and less advanced pathologic stages. PMID- 16360235 TI - Quality of life and cost-effectiveness of anti-HCV therapy in HIV-infected patients. AB - Quality of life studies in chronic hepatitis C using specific questionnaires have been performed in mono-infected patients; however, these studies have just begun in HIV-HCV co-infected patients. The questionnaires used in mono-infected patients are not well adapted for co-infected patients and need to be redesigned. Typically, co-infected patients have multiple symptoms that may be attributable to HIV, to HCV, to a combination of both diseases or even to side effects related to drug therapy. These patients usually need substitutive therapy to manage side effects related to HCV therapy, particularly anaemia, leukopenia and depression. There are no cost-effectiveness studies published on the current HCV standard therapy with pegylated interferon and ribavirin. However, there is some research on the old standard interferon and ribavirin, which shows that HCV therapy is cost-effective. Cost-effectiveness studies in co-infected patients will have to take into account variables that do not affect mono-infected patients, such as the different levels of CD4, the increase in the fibrosis progression rate and the use of other expensive drugs for the management of side effects. Currently, the literature does not provide adequate information on the effect of HCV infection on the quality of life of HIV-HCV co-infected patients or the most cost effective HCV therapy. PMID- 16360236 TI - Developing a marketing function in public healthcare systems: a framework for action. AB - The scope of this paper is to analyse the contribution that a marketing function can bring to the wide variety of healthcare organizations operating in public health systems (PHSs). While extensive research on marketing applied to healthcare services has been elaborated in competitive and managed care contexts, marketing is a rather new issue in PHSs and little research has been conducted to assess its relevance and benefits in these environments. This study tackles that gap and is based on a review of the current literature in order to provide answers to the following points: - definition of the scopes of marketing and of the elements that affect its incorporation in the healthcare sector; - conceptualization of the possible approaches to marketing by health organizations operating in PHSs; - discussion of the resulting framework for action. PMID- 16360237 TI - Pemetrexed-associated urticarial vasculitis. AB - Pemetrexed (Alimta, Eli Lilly and Company, Indianapolis, IN) is a structurally novel anti-folate agent. The United States Food and Drug Administration has approved pemetrexed for the treatment of patients with malignant pleural mesothelioma and previously treated patients with locally advanced or metastatic non-small cell lung cancer. In the phase III trials that led to its approval, rash was reported in 17 and 22% of patients receiving pemetrexed alone or in combination with cisplatin. However, little has been published about the characteristics of this rash or about its mechanism. In an attempt to contribute to the growing body of knowledge about this new agent, we describe a case in which a patient developed a rash secondary to urticarial vasculitis associated with pemetrexed. PMID- 16360238 TI - Second-line treatment for advanced non-small cell lung cancer: a systematic review. AB - BACKGROUND: Among advanced non-small cell lung cancer (NSCLC) patients, most will resist or relapse after first-line chemotherapy. As a result, second-line therapy has been a major focus for clinical research. MATERIALS AND METHODS: A systematic review was carried out from 1996 to February 2005. RESULTS: Second-line chemotherapy provides pre-treated NSCLC patients with a clear survival advantage. Docetaxel 75 mg/m(2) every 3 weeks is the present standard second-line chemotherapy. Despite promising results regarding efficacy and toxicity in phase III studies, a docetaxel weekly schedule could not be recommended. Pemetrexed recently emerged as an alternative with similar efficacy and less toxicity. Although the combination of two drugs was not associated with a survival benefit when compared with single-agent chemotherapy, such regimens induced a dramatic increase in toxicities and therefore mono-chemotherapy remains the standard as second-line therapy. Finally, few new agents were reported with better results than those used previously and clinical research on second-line therapy currently focuses on combinations with targeted therapies. CONCLUSION: Second-line chemotherapy offers NSCLC patients a small but significant survival improvement. However, this field of clinical research needs further investigations in order to answer certain remaining questions especially concerning targeted therapies. PMID- 16360239 TI - Steam stripping of the unsaturated zone of contaminated sub-soils: the effect of diffusion/dispersion in the start-up phase. AB - The unsteady process of steam stripping of the unsaturated zone of soils contaminated with volatile organic compounds (VOCs) is addressed. A model is presented. It accounts for the effects of water and contaminants remaining in vapour phase, as well as diffusion and dispersion of contaminants in this phase. The model has two components. The first is a one-dimensional description of the propagation of a steam front in the start-up phase. This is based on Darcy's law and conservation laws of mass and energy. The second component describes the transport of volatile contaminants. Taking the view that non-equilibrium between liquid and vapour phases exists, it accounts for evaporation, transport, and condensation at the front. This leads to a moving-boundary problem. The moving boundary problem is brought into a fixed domain by a suitable transformation of the governing partial differential equations, and solved numerically. For a broad range of the governing dimensionless numbers, such as the Henry, Merkel and Peclet numbers, computational results are discussed. A mathematical asymptotic analysis supports this discussion. The range of parameter values for which the model is valid is investigated. Diffusion and dispersion are shown to be of qualitative importance, but to have little quantitative effect in the start-up phase. PMID- 16360240 TI - Assembly and mechanosensory function of focal adhesions: experiments and models. AB - Initial integrin-mediated cell-matrix adhesions (focal complexes) appear underneath the lamellipodia, in the regions of the "fast" centripetal flow driven by actin polymerization. Once formed, these adhesions convert the flow behind them into a "slow", myosin II-driven mode. Some focal complexes then turn into elongated focal adhesions (FAs) associated with contractile actomyosin bundles (stress fibers). Myosin II inhibition does not suppress formation of focal complexes but blocks their conversion into mature FAs and further FA growth. Application of external pulling force promotes FA growth even under conditions when myosin II activity is blocked. Thus, individual FAs behave as mechanosensors responding to the application of force by directional assembly. We proposed a thermodynamic model for the mechanosensitivity of FAs, taking into account that an elastic molecular aggregate subject to pulling forces tends to grow in the direction of force application by incorporating additional subunits. This simple model can explain a variety of processes typical of FA behavior. Assembly of FAs is triggered by the small G-protein Rho via activation of two major targets, Rho associated kinase (ROCK) and the formin homology protein, Dia1. ROCK controls creation of myosin II-driven forces, while Dia1 is involved in the response of FAs to these forces. Expression of the active form of Dia1, allows the external force-induced assembly of mature FAs, even in conditions when Rho is inhibited. Conversely, downregulation of Dia1 by siRNA prevents FA maturation even if Rho is activated. Dia1 and other formins cap barbed (fast growing) ends of actin filaments, allowing insertion of the new actin monomers. We suggested a novel mechanism of such "leaky" capping based on an assumption of elasticity of the formin/barbed end complex. Our model predicts that formin-mediated actin polymerization should be greatly enhanced by application of external pulling force. Thus, the formin-actin complex might represent an elementary mechanosensing device responding to force by enhancement of actin assembly. In addition to its role in actin polymerization, Dia1 seems to be involved in formation of links between actin filaments and microtubules affecting microtubule dynamics. Alpha-tubulin deacetylase HDAC6 cooperates with Dia1 in formation of such links. Since microtubules are known to promote FA disassembly, the Dia1 mediated effect on microtubule dynamics may possibly play a role in the negative feedback loop controlling size and turnover of FAs. PMID- 16360241 TI - The molecular dynamics of osteoclast adhesions. AB - Podosomes are specialized adhesive structures that play a central role in bone resorption. In this article we address the molecular diversity and dynamics of podosomes at different states of organization, ranging from scattered distribution over the entire ventral membrane of non-polarized cells, via formation of podosome clusters and developing rings to the assembly of a peripheral belt, resembling the sealing zone of polarized, bone-resorbing osteoclasts. Based on published data and on our own results, we describe here the spatial relationships between key podosome-associated proteins. Using quantitative microscopy, we show here a dramatic increase in the local levels of F-actin, vinculin, paxillin, and alpha-actinin, which occurs upon the transformation of clustered podosomes into rings and sealing zone-like structures. This change is accompanied by a marked decrease in phosphotyrosine levels in the same region. Therefore, our data suggest that a major change in the molecular composition of podosomes is taking place during osteoclast polarization, a change that may be related to adhesion "reinforcement", associated with the assembly of the bone-resorbing apparatus. Studying the nature of the proteins that undergo de-phosphorylation is critical for the understanding of the mechanisms regulating the processes described above. PMID- 16360242 TI - [Drugs induced Parkinsonism: the case of a 77 year old woman]. PMID- 16360243 TI - Modelling the spatio-temporal evolution of 3H in the waters of the River Tagus. AB - Measurements of tritium specific activity levels and of temperatures in waters of the Torrejon-Tagus reservoir (Spain) showed that their radioactive characteristics were basically influenced by the radioactive liquid effluent from the Almaraz Nuclear Power Plant. This enters the Torrejon-Tagus reservoir via the Arrocampo cooling reservoir, which exchanges water with the first. We studied the temporal and spatial (in two dimensions) evolution of the mentioned parameters for years 1997 and 1998. The tritium levels were found to be significantly correlated with temperature. Two numerical models were constructed for a quantitative study of the tritium levels along Torrejon reservoir: a 1D model was used for the dispersion of tritium along the whole length of the reservoir, and a 2D depth-averaged model was used for a detailed study of the area where tritium is released into the reservoir. Both models solve the hydrodynamic equations, to obtain the currents induced by the exchanges of water between the reservoirs in the River Tagus and Arrocampo, and the advection/diffusion equation to calculate the dispersion of tritium. In general, the model results were in agreement with the experimental observations. PMID- 16360244 TI - Pyoderma gangrenosum--case report. AB - We report the case of a woman of 34 years who had ulcerative colitis and atypical pyoderma gangrenosum. The pyoderma gangrenosum responded to conservative treatment. PMID- 16360245 TI - Influence of NICE guidelines on removal of third molars in a region of the UK. AB - We investigated compliance in the West Midlands, UK with the guidelines issued by the National Institute of Clinical Excellence (NICE) on the indications for the removal of third molars. We audited the practice at three departments of maxillofacial surgery that take referrals from three districts of the West Midlands. Of a total of 300 referral letters 217 (72%) specified a NICE-compliant diagnosis. A total of 267 patients (89%) were advised to have third molars removed and all three units achieved 100% compliance with NICE guidelines. There was a concordance of diagnosis of 69% between referring general dental practitioners and attending oral surgeons (concordance was 79% if only those referral letters that carried a specific indication were considered). Disease free wisdom teeth made up a mean of only 6% of all referrals. PMID- 16360246 TI - Transfusion-related acute lung injury. AB - Transfusion-related acute lung injury (TRALI) is a life-threatening adverse event of transfusion, which has an increasing incidence in the United States and is the leading cause of transfusion-related death. TRALI and acute lung injury (ALI) share a common clinical definition except that TRALI is temporally- and mechanistically-related to transfusion of blood or blood components. A number of different models have been proposed to explain the pathogenesis. The first is an antibody-mediated event whereby transfusion of anti-HLA, class I or class II, or anti-granulocyte antibodies into patients whose leukocytes express the cognate antigens. The antibody:antigen interaction causes complement-mediated pulmonary sequestration and activation of neutrophils (PMNs) resulting in TRALI. The second is a two-event model: the first event is the clinical condition of the patient resulting in pulmonary endothelial activation and PMN sequestration, and the second event is the transfusion of a biologic response modifier (including anti granulocyte antibodies, lipids, and CD40 ligand) that activates these adherent PMNs resulting in endothelial damage, capillary leak, and TRALI. These hypotheses are discussed with respect to animal models and human studies that provide the experimental and clinical relevance. The definition of TRALI, patient predisposition, treatment, prevention and reporting guidelines are also examined. PMID- 16360247 TI - Characterization of contamination in and toxicities of a shipyard area in Hong Kong. AB - This is the first integrated study on the contaminant characterization and toxicities of a shipyard area. This site in Hong Kong contained 3.6-33.4 mg kg( 1) dichlorodiphenyltrichloroethane and 5-79 g kg(-1) oil and grease. Other pollutants exceeding the Dutch intervention values were total polyaromatic hydrocarbons, total petroleum hydrocarbons, and metals Cu, Pb and Zn. These pollutants persisted partially owing to the poor fertility with <125 mg N kg(-1) and <530 mg P kg(-1) typical of coastal soil. Sediments collected within 100 m also showed contamination exceeding the China and Hong Kong intervention levels. The shipyard soil killed two bacteria Pseudomonas stutzeri and Acinetobacter sp. isolated from a clean coastal soil, inhibited germination of three exogenous plants Triticum aestivum, Brassica chinense and Lolium perenne, and inhibited proliferation of three indigenous bacteria: Methylobacterium sp., Pseudomonas aeruginosa and Bacillus cereus. Thus, ex situ remediation was needed. PMID- 16360248 TI - Geostatistical analysis and kriging of Hexachlorocyclohexane residues in topsoil from Tianjin, China. AB - A previously published data set of HCH isomer concentrations in topsoil samples from Tianjin, China, was subjected to geospatial analysis. Semivariograms were calculated and modeled using geostatistical techniques. Parameters of semivariogram models were analyzed and compared for four HCH isomers. Two dimensional ordinary block kriging was applied to HCH isomers data set for mapping purposes. Dot maps and gray-scaled raster maps of HCH concentrations were presented based on kriging results. The appropriateness of the kriging procedure for mapping purposes was evaluated based on the kriging errors and kriging variances. It was found that ordinary block kriging can be applied to interpolate HCH concentrations in Tianjin topsoil with acceptable accuracy for mapping purposes. PMID- 16360249 TI - Reproductive transfer of organochlorines in viviparous surfperch, Ditrema temmincki. AB - To discuss the maternal-fetal transfer of persistent organic pollutants (POPs) in viviparous fish, the whole-body burdens of POPs were determined in gravid surfperches from Otsuchi Bay, Japan, and in their fetuses. The concentrations were 2.75-15.5 times higher in the maternal tissue. To compare the characteristics of the maternal-fetal transfer between species with different gestation systems, reports on the striped dolphin and long-finned pilot whales were studied. The transfer rates of POPs in surfperches were 0.6-6.0%, while in marine mammals they were 3.0-9.4%. The difference in transfer rates may derive from the difference in their reproductive systems. On the other hand, as in marine mammals, selective transfer of POPs was seen in surfperches. The transfer rates were higher in less hydrophobic compounds such as dieldrin, and lower in more hydrophobic compounds such as DDTs. A new transfer factor is also discussed which instantly indicates the difference in the concentrations between mother and fetus. PMID- 16360250 TI - Plant-mediated effects of heavy metal pollution on host choice of a grass miner. AB - We studied the effects of heavy metal exposure on host plant choice and performance of the grass miner Chromatomyia milii (Diptera, Agromyzidae). Cadmium decreased plant growth in a dose-dependent way. C. milii preferred the control to the cadmium-exposed plants for feeding and oviposition. Moreover, preference for the control plants increased with increasing cadmium exposure of the alternative choice. Adult and offspring performance decreased with increasing plant cadmium exposure. This suggests that, at least under our laboratory conditions, host choice of C. milii is adaptive under pollution stress. Foliar cadmium concentration increased and the soluble sugar concentration decreased with increasing cadmium exposure. Regression analysis showed that both latter components might be responsible for the decrease in performance of C. milii on cadmium-exposed plants. The protein and amino acid concentration of the leaves, the amount of structural defenses, and water concentration were not affected by the cadmium treatment. PMID- 16360251 TI - Polycyclic aromatic hydrocarbons affect survival and development of common snapping turtle (Chelydra serpentina) embryos and hatchlings. AB - Polycyclic aromatic hydrocarbons (PAHs) are toxic compounds found in the John Heinz National Wildlife Refuge in Philadelphia, Pennsylvania. We assessed the impact of PAHs and crude oil on snapping turtle development and behavior by exposing snapping turtle eggs from the Refuge and from three clean reference sites to individual PAHs or a crude oil mixture at stage 9 of embryonic development. Exposure to PAHs had a significant effect on survival rates in embryos from one clean reference site, but not in embryos from the other sites. There was a positive linear relationship between level of exposure to PAHs and severity of deformities in embryos collected from two of the clean reference sites. Neither righting response nor upper temperature tolerance (critical thermal maximum, CTM) of snapping turtle hatchlings with no or minor deformities was significantly affected by exposure to PAHs. PMID- 16360252 TI - The marine macroalga Cystoseira baccata as biosorbent for cadmium(II) and lead(II) removal: kinetic and equilibrium studies. AB - This work reports kinetic and equilibrium studies of cadmium(II) and lead(II) adsorption by the brown seaweed Cystoseira baccata. Kinetic experiments demonstrated rapid metal uptake. Kinetic data were satisfactorily described by a pseudo-second order chemical sorption process. Temperature change from 15 to 45 degrees C showed small variation on kinetic parameters. Langmuir-Freundlich equation was selected to describe the metal isotherms and the proton binding in acid-base titrations. The maximum metal uptake values were around 0.9 mmol g(-1) (101 and 186 mg g(-1) for cadmium(II) and lead(II), respectively) at pH 4.5 (raw biomass), while the number of weak acid groups were 2.2 mmol g(-1) and their proton binding constant, K(H), 10(3.67) (protonated biomass). FTIR analysis confirmed the participation of carboxyl groups in metal uptake. The metal sorption was found to increase with the solution pH reaching a plateau above pH 4. Calcium and sodium nitrate salts in solution were found to affect considerably the metal biosorption. PMID- 16360253 TI - High incidence of deformity in aquatic turtles in the John Heinz National Wildlife Refuge. AB - The John Heinz National Wildlife Refuge is subject to pollution from multiple sources. We studied development of snapping turtle (Chelydra serpentina) and painted turtle (Chrysemys picta) embryos from the refuge from 2000 through 2003. Mean annual deformity rate of pooled painted turtle clutches over four years ranged from 45 to 71%, while that of snapping turtle clutches ranged from 13 to 19%. Lethal deformities were more common than minor or moderate deformities in embryos of both species. Adult painted turtles had a higher deformity rate than adult snapping turtles. Snapping turtles at JHNWR had high levels of PAH contamination in their fat. This suggests that PAHs are involved in the high level of deformities. Other contaminants may also play a role. Although the refuge offers many advantages to resident turtle populations, pollution appears to place a developmental burden on the life history of these turtles. PMID- 16360254 TI - Physicochemical characterization and mercury speciation of particle-size soil fractions from an abandoned mining area in Mieres, Asturias (Spain). AB - Soils from old cinnabar mining areas usually exhibit high Hg contents, whose mobility depends on soil parameters and environmental conditions. This paper presents the study of the Hg speciation in soil samples from an abandoned Hg mine and metallurgical plant in Mieres (Asturias, Spain), in relation to their mineralogical and chemical composition and their particle-size distribution. A characterization of samples was made by X-Ray Diffraction Spectrometry, Scanning Electron Microscopy and Atomic Absorption and Emission Spectroscopy analyses. A sequential extraction method was applied to establish Hg mobility in the samples and their grain-size subsamples. The highest Hg mobility was found in well developed soils, as a consequence of the adsorption processes by iron and manganese oxides, whereas in those more contaminated soils, a higher proportion of Hg was leached in the non-mobile fraction. A higher Hg mobility was found in the finest grain-size subsamples, probably due to the accumulation of clay minerals and oxides in these ranges. PMID- 16360255 TI - Relapse in the addictive behaviors: integration and future directions. AB - This paper identifies the major consistencies in substantive and methodological findings across the review papers in this special issue on relapse in the addictive behaviors. The papers were consistent in suggesting that there have been major methodological advances which have helped to move the field forward. Furthermore, the papers show the need for taking a biopsychosocial approach to the study of relapse and the major difficulty across addictive behaviors in creating an acceptable operational definition of relapse. Suggestions for future research directions that follow from the papers include deriving and evaluating relapse definitions, systematically developing and testing models and theories of relapse, and understanding and narrowing the relapse research-clinical practice gap. PMID- 16360256 TI - A review of the reference dose for chlorpyrifos. AB - Chlorpyrifos is an inhibitor of cholinesterase (ChE) and inhibition of ChE is believed to be the most sensitive effect in all animal species evaluated and in humans from previous evaluations. Recent literature, in particular epidemiology studies reporting associations between chlorpyrifos levels and fetal birth weight decreases, suggest the need to reevaluate the basis of the reference dose (RfD) for chlorpyrifos, however. In this paper, we evaluated newly available publications regarding chlorpyrifos toxicity and discuss the choice of critical effect--whether cholinesterase inhibition or developmental effect, the choice of appropriate species and study, the appropriate point of departure, and choice of uncertainty factors--including a discussion of the FQPA safety factor. We conclude that RBC cholinesterase inhibition is the critical effect, that human studies form the best choice of species--supported by a wealth of experimental animal data, that a NOAEL of 0.1 mg/kg/day is the most appropriate point of departure, and that a 10-fold factor for within human variability is sufficient to characterize the overall uncertainty in this rather large database. The resulting RfD is 0.01 mg/kg/day. PMID- 16360257 TI - Infertility: testing a helpseeking model. AB - This paper uses data from a study of 196 infertile women from the Midwestern US to examine a general theory of helpseeking behavior applied to infertility. All of these women report meeting the medical definition of infertility--12 months or more of regular intercourse without conception--at some point in their lives. Only 35 percent of this sample of infertile women identified themselves as having had fertility problems and only 40 percent had sought medical treatment. Drawing on prior theories of helpseeking, we examine the effects of symptom salience, life course cues, attitudes, predisposing factors, and enabling conditions on helpseeking. We posit a model in which a cognitive dimension (perceived infertility) mediates between these predictors and medical helpseeking. Symptom salience (experienced infertility while actively trying to get pregnant), low parity, and poor subjective health are significantly related to perceived infertility, which is, in turn, significantly associated with helpseeking for infertility. Supporting the conclusion that the cognitive dimension of identifying oneself as infertile is critical to helpseeking, the relationship of symptom salience to helpseeking is partially mediated by perceived problems. Internal health locus of control is associated with lower odds of helpseeking but not to perceived infertility. PMID- 16360258 TI - Whey protein concentrate promotes the production of glutathione (GSH) by GSH reductase in the PC12 cell line after acute ethanol exposure. AB - Excessive ethanol consumption may increase the production of reactive oxygen species (ROS), which results in the damage of tissues, especially the neurons and glial cells in the central nervous system (CNS). The purpose of this study is to evaluate the effects of whey protein concentrate (WPC) on the glutathione (GSH) status after acute ethanol exposure in the pheochromocytoma (PC12) cell line. In this study, we assayed the cell viability, the percentage of lactate dehydrogenase released (% LDH released), the level of GSH, and the activity of GSH reductase (GRx). The results showed that with the supplement of WPC, the cell viability displayed no significant difference after acute exposure of ethanol in groups with or without ethanol treatment. The ethanol-induced cytotoxicity showed a slight decrease ,and the level of GSH showed a significant increase. The activity of GRx significantly increased when 0.1, 10mg/ml of WPC was supplied. In conclusion, these results suggest that WPC in a moderate concentration should be a precursor agent to promote the production of GSH and will enhance the antioxidant capacity in the PC12 cell line. PMID- 16360259 TI - alpha2,3/alpha2,6-Sialylation of N-glycans: non-synonymous signals with marked developmental regulation in bovine reproductive tracts. AB - The glycan part endows cellular glycoconjugates with significant potential for biological recognition. N-Glycan branches often end with alpha2,3/alpha2,6 sialylation, posing the question whether and how placement of the sialic acid at 3 - or 6 -acceptor positions of galactose has cell biological relevance. As attractive model to study developmental regulation we monitored the expression of alpha2,3/alpha2,6-sialylated determinants in fetal and adult bovine testes and ovaries by lectin histochemistry. Distinct expression patterns were detected in both organ types. Oocyte staining, as a prominent example, was restricted to the presence of alpha2,6-sialylated glycans. Treatment with sialidase abolished binding and thus excluded sulfate esters as lectin targets. We added computer simulations to rationalize the observed evidence for non-random expression of the two closely related sialylgalactose isomers. Extensive molecular mechanics and molecular dynamics calculations reveal that the seemingly minor shift of the glycosidic bond from the alpha2,3 position to the alpha2,6 configuration causes significant shape and flexibility changes. They give each disaccharide its own characteristic meaning as signal in the sugar code. PMID- 16360260 TI - The validity of oral glucose tolerance test after 36 weeks' gestation. AB - OBJECTIVE: Secretion of anti-insulin hormones plateaus near term, questioning the validity of OGTT (oral glucose tolerance test) during that period. We aimed at assessing the feasibility of OGTT near term as compared to OGTT at 26-32 weeks. PATIENTS AND METHODS: One thousand four hundred and eighty seven pregnant women were screened by GCT (glucose challenge test), and 282 (19%) of them performed an OGTT at 26th-32nd weeks ("early" OGTT) after meeting the threshold value for GCT. Forty-one women with abnormal and 16 with normal early OGTT underwent a repeated OGTT at 36-40 weeks' gestation ("late" OGTT). Blood glucose levels during GCT and OGTT were compared between women with early and late abnormal OGTT and women who converted from early abnormal to late normal OGTT. RESULTS: Thirty-six out of 41 participants (88%) with early abnormal OGTT had abnormal test near term as well (Group I). Five women with an early abnormal OGTT converted to normal according to a late OGTT (Group II). These women had lower glucose levels on both late and early OGTT as compared with Group I. All 16 women who tested normal on early OGTT had a consistently normal late OGTT. Glucose levels for all 57 women did not significantly differ between early and late OGTT. The sensitivity, specificity, and positive and negative predictive values of late OGTT were 88%, 100%, 100%, and 76%, respectively. CONCLUSION: The positive predictive value of late OGTT performed at 36-40 weeks' gestation is 100%. This test may be used to detect gestational diabetes in women near term. PMID- 16360261 TI - Induction of labor and perinatal outcome: the impact of the amniotic fluid index. AB - OBJECTIVE: The purpose was to determine the impact of the amniotic fluid index on the perinatal outcome of patients admitted for induction of labor at term. STUDY DESIGN: Patients (n=180) included in the study were those admitted for induction of labor at 37-42 weeks' gestation, with unfavorable cervix and intact membranes. The amniotic fluid index (AFI) was determined in all patients using the four quadrant technique within 24 h of the induction of labor. Patients were divided into two groups based on their AFI: the oligohydramnios group with AFI < or = 5 cm (n = 66) and a normal group with AFI > 5 cm (n = 114). The perinatal outcomes of the two groups were compared. RESULTS: The two groups were similar with regard to maternal age, gestational age, and birth weight. Meconium staining of the amniotic fluid was significantly higher in the group with AFI < or = 5 cm (p = 0.040). The number of cesarean deliveries due to fetal distress was significantly higher even after adjusting for other confounding factors in the group with AFI < or = 5 cm (adjusted OR 6.52 [95% CI 1.82, 23.2]; p < or = 0.0001). There was no significant difference between the two groups with regard to Apgar scores or neonatal admission. CONCLUSION: Induction of labor at term in patients with oligohydramnios is associated with an increased risk of cesarean delivery due to fetal distress. PMID- 16360262 TI - Metformin versus rosiglitazone in the treatment of polycystic ovary syndrome. AB - OBJECTIVE: The aim of our study is to investigate and compare the clinical, biochemical and hormonal changes during application of insulin-sensitizers from two different groups. STUDY DESIGN: This prospective, open clinical study lasted 3 months and included 30 women with PCOS, divided in two groups of 15 women each. Group 1 received 850 mg metformin twice a day and group 2 was treated with rosiglitazone 4 mg a day. Serum levels of testosterone, immune reactive insulin (IRI), sex hormone binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEAS) and lipid metabolism parameters were measured before the treatment, and on the 3rd month. Free androgen index (FAI) and homeostasis model assessment of insulin resistance (HOMA-IR) were calculated. Body mass index (BMI) and waist-to hip ratio (WHR) were assessed at baseline and at the end of therapy. RESULTS: Two parameters change significantly in the 3rd month in our study--testosterone and insulin. Much better decrease in the level of testosterone and free androgen index was established in group treated with metformin, while the indices of insulin resistance were better influenced in the group treated with rosiglitazone. CONCLUSION: Application of insulin sensitizers from both groups has a favorable influence on the basic hormonal deviations in PCOS--the hyperandrogenemia and the insulin resistance. In cases with PCOS metformin treatment influences better hyperandrogenemia, while rosiglitazone affects more pronouncedly insulin resistance and hyperinsulinemia. PMID- 16360263 TI - Adsorption of acid dye onto organobentonite. AB - Removal of Acid Red 151 from aqueous solution at different dye concentrations, adsorbent doses and pH has been studied. The bentonite clay has been modified using cationic surfactants, which has been confirmed using XRD and FT-IR analyses. Experimental result has shown that the acidic pH favours the adsorption. The adsorption isotherms are described by means of Langmuir and Freundlich isotherms. The adsorption capacity has been found to be 357.14 and 416.66 mg g(-1) for the cetyldimethylbenzylammonium chloride-bentonite (CDBA bent) and cetylpyridinium chloride-bentonite (CP-bent), respectively. Kinetic studies show that the adsorption followed second-order kinetics. PMID- 16360264 TI - Database management systems for process safety. AB - Several elements of the process safety management regulation (PSM) require tracking and documentation of actions; process hazard analyses, management of change, process safety information, operating procedures, training, contractor safety programs, pre-startup safety reviews, incident investigations, emergency planning, and compliance audits. These elements can result in hundreds of actions annually that require actions. This tracking and documentation commonly is a failing identified in compliance audits, and is difficult to manage through action lists, spreadsheets, or other tools that are comfortably manipulated by plant personnel. This paper discusses the recent implementation of a database management system at a chemical plant and chronicles the improvements accomplished through the introduction of a customized system. The system as implemented modeled the normal plant workflows, and provided simple, recognizable user interfaces for ease of use. PMID- 16360265 TI - Activation of protein kinase B/Akt signaling pathway contributes to mechanical hypersensitivity induced by capsaicin. AB - We investigated the involvement of the protein kinase B/Akt (PKB/Akt) signaling pathway in the mechanical hypersensitivity induced in rats by capsaicin. Intradermal injection of capsaicin results in activation of PKB/Akt in the lumbar spinal cord, most prominently in the dorsal horn, starting by 5 min after capsaicin injection and lasting at least 1h. The activated PKB/Akt in the spinal cord is in neurons, since phospho-PKB/Akt (p-PKB/Akt) colocalizes with the neuronal marker, neuronal-specific nuclear protein (NeuN). The mechanical hypersensitivity is shown by the enhanced paw withdrawal frequency to applications of von Frey filaments with different bending forces (30, 100, 200 mN) on the rat paw. Pre-treatment with several different PKB/Akt inhibitors, including SH-6, Akt inhibitor IV, and Akt inhibitor V, blocked the mechanical hypersensitivity induced by intradermal injection of capsaicin, a measure of spinal cord central sensitization. Two structurally unrelated phosphoinositide 3 Kinase (PI3K, upstream of PKB/Akt) inhibitors, Wortmannin and LY294002, also prevented the mechanical hypersensitivity induced by intradermal injection of capsaicin. Furthermore, post-treatment with the PI3K inhibitor, Wortmannin, or PKB/Akt inhibitors, such as NL-71-101, SH-6, Akt inhibitor IV, and inhibitor V significantly reduced the established mechanical hypersensitivity induced by capsaicin. The PKB/Akt signaling pathway in the spinal cord is therefore involved in pain hypersensitivity. PMID- 16360266 TI - Transcutaneous electrical nerve stimulation (TENS) reduces chronic hyperalgesia induced by muscle inflammation. AB - Transcutaneous electrical nerve stimulation (TENS) reduces pain through central mechanisms involving spinal cord and brainstem sites. Since TENS acts through central mechanisms, we hypothesized that TENS will reduce chronic bilateral hyperalgesia produced by unilateral inflammation when applied either ipsilateral or contralateral to the site of muscle inflammation. Sprague-Dawley rats were injected with carrageenan in the left gastrocnemius muscle belly. Mechanical withdrawal threshold was tested bilaterally before and 2 weeks after carrageenan injection. After testing withdrawal thresholds at 2 weeks, rats received TENS treatment either ipsilateral or contralateral to the site of inflammation. In each of these groups, rats were randomized to control (no TENS), low frequency (4 Hz), or high frequency (100 Hz) TENS treatment. TENS was applied for 20 min at sensory intensity under light halothane anesthesia. Mechanical withdrawal thresholds were re-assessed after TENS or 'no TENS' treatment. Unilateral injection of carrageenan to the gastrocnemius muscle significantly reduced the mechanical withdrawal threshold (mechanical hyperalgesia) bilaterally 2 weeks later. Either low or high frequency TENS applied to the gastrocnemius muscle ipsilateral to the site of inflammation significantly reversed mechanical hyperalgesia, both ipsilateral and contralateral to the site of inflammation. Low or high frequency TENS applied to the gastrocnemius muscle contralateral to the site of inflammation also significantly reduced mechanical hyperalgesia, both ipsilateral and contralateral to the site of inflammation. Since ipsilateral or contralateral TENS treatments were effective in reducing chronic bilateral hyperalgesia in this animal model, we suggest that TENS act through modulating descending influences from supraspinal sites such as rostral ventromedial medulla (RVM). PMID- 16360267 TI - Increased taste intensity perception exhibited by patients with chronic back pain. AB - There is overlap between brain regions involved in taste and pain perception, and cortical injuries may lead to increases as well as decreases in sensitivity to taste. Recently it was shown that chronic back pain (CBP) is associated with a specific pattern of brain atrophy. Since CBP is characterized by increased sensitivity to pain, we reasoned that the sense of taste might also be enhanced in CBP. Detection and recognition thresholds were established for a sour taste and ratings of both suprathreshold taste intensity and pleasantness unpleasantness perception were collected for sweet, sour, salty and bitter stimuli in 11 CBP patients and 11 matched control subjects. As a control, ratings were also collected for visual assessment of degree of grayness. There was no difference between CBP and control subjects for visual grayness rating. On the other hand, CBP patients in comparison to control subjects rated gustatory stimuli as significantly more intense but no more or less pleasant and showed a trend towards a lower detection threshold (i.e. increased sensitivity). The selectivity of the taste disturbance suggests interaction between pain and taste at specific brain sites and provides further evidence that CBP involves specific brain abnormalities. PMID- 16360268 TI - Deep brain stimulation for chronic pain: can it help? PMID- 16360269 TI - Spinal injections: getting hold of the wrong end of the stick. PMID- 16360270 TI - GW274150, a novel and highly selective inhibitor of the inducible isoform of nitric oxide synthase (iNOS), shows analgesic effects in rat models of inflammatory and neuropathic pain. AB - Nitric oxide (NO), synthesised by different isoforms of nitric oxide synthase (NOS), has been linked with the development and maintenance of nociception. We studied the role of the inducible isoform, iNOS, in two different rat pain models with an inflammatory component. iNOS was immunohistochemically detected locally in the paw 6h after Freund's Complete Adjuvant (FCA) injection, showing a plateau at 24-72 h and falling slowly in the following weeks. This correlated with the late phase of the hypersensitivity to pain revealed in the behavioural tests. A highly selective iNOS inhibitor GW274150 (1-30 mg/kg orally, 24h after FCA) suppressed the accumulation of nitrite in the inflamed paw indicating substantial iNOS inhibition. At the same time it partially reversed FCA-induced hypersensitivity to pain and edema in a dose-dependent manner. After Chronic Constriction Injury (CCI) surgery to the sciatic nerve, iNOS presence was only detected locally in the region of the nerve (inflammatory cells). GW274150 (3-30 mg/kg orally, 21 days after surgery) also reversed significantly the CCI associated hypersensitivity to pain. No iNOS was detectable in dorsal root ganglia, spinal cord or brain in either model. This study demonstrates a role for peripherally-expressed iNOS in pain conditions with an inflammatory component and the potential value of iNOS inhibitors in such conditions. PMID- 16360271 TI - Persistence of low back pain reporting among a cohort of employees in a metal corporation: a study with 5-, 10-, and 28-year follow-ups. AB - Low back pain (LBP) is a common symptom among adults but little is known about its persistence over time in defined populations. The aim of this study was to examine the persistence of LBP among a cohort of industrial employees studied in four successive surveys during a total of 28 years. Cross-tabulations and logistic regression was used to estimate the interdependence of LBP occurrence at the surveys. At baseline, 54% of the subjects reported local LBP and 25% LBP radiating to the lower limb(s). Persistent or recurrent LBP was common. Of those with LBP at baseline, 75, 73, and 88% reported it also at the 5-, 10- or 28-year follow-up, respectively. Of those with radiating pain, 66, 65, and 69% were symptomatic 5, 10, or 28 years later. The onset of reporting LBP increased during follow-up. Of those without local LBP at baseline, 33, 37 and 64% had pain at the 5-, 10-, or 28-year follow-up, respectively. Of those without radiating LBP, 17, 22, and 46% had pain at the 5-, 10-, or 28-year follow-up. The odds ratio of local LBP at the 5-, 10-, or 28-year follow-up for those with such pain at baseline vs. not were 6.0 (95% CI 4.3-8.3), 4.7 (3.3-6.6) and 4.0 (2.6-6.3), adjusted for age, gender and occupational class. The respective figures for radiating LBP were 8.5 (5.7-12.5), 6.7 (4.4-10.1) and 2.3 (1.5-3.6). We conclude that LBP is commonly recurrent. PMID- 16360272 TI - Modulation of P2X receptors via adrenergic pathways in rat dorsal root ganglion neurons after sciatic nerve injury. AB - The present study examined noradrenaline-induced modulation of ATP-evoked currents in dorsal root ganglion (DRG) neurons after sciatic nerve injury (transection). ATP (10 microM) generated fast/mixed type of whole-cell membrane currents, possibly as mediated via P2X(3)/P2X(3)-like receptors, and slow type of the currents, possibly as mediated via P2X(2/3) receptors, in acutely dissociated L4/5 DRG neurons, without significant difference between sham and injury group. For sham group, noradrenaline (10 microM) enhanced fast/mixed type of ATP-evoked currents in ipsilateral DRG neurons, that is not inhibited by H-7, a broad inhibitor of protein kinases, but otherwise it had no effect on slow type of the currents. For injury group, noradrenaline (10 microM) significantly potentiated slow type of ATP-evoked currents in ipsilateral DRG neurons, that is abolished by H-7 or GF109203X, a selective inhibitor of protein kinase C (PKC), while it depressed fast/mixed type of the currents. In the analysis of real-time reverse transcription-polymerase chain reaction, an increase in the mRNAs for alpha(1b), alpha(2a), alpha(2d), and beta(2) adrenergic receptors was found with the ipsilateral DRGs after sciatic nerve injury. Collectively, the results of the present study suggest that noradrenaline potentiates P2X(2/3) receptor currents by activating PKC via alpha(1) adrenergic receptors linked to G(q) protein, perhaps dominantly alpha(1b) adrenergic receptors, in DRG neurons after sciatic nerve injury. This may account for a nociceptive pathway in response to noradrenergic sprouting after peripheral nerve injury. PMID- 16360273 TI - Downregulation of spinal glutamate transporter EAAC1 following nerve injury is regulated by central glucocorticoid receptors in rats. AB - Previous studies have shown that glucocorticoid receptors (GR) were upregulated, whereas glutamate transporters were downregulated, within the spinal cord dorsal horn after peripheral nerve injury. However, the relationship between the expression of spinal GR and glutamate transporter after nerve injury remains unknown. In the present study, we examined the hypothesis that central GR would regulate the expression of spinal glutamate transporter EAAC1 following chronic constriction nerve injury (CCI) in rats. CCI induced a significant downregulation of EAAC1 expression primarily within the ipsilateral spinal cord dorsal horn when examined on postoperative day 7 using both Western blot and immunohistochemistry. The downregulation of EAAC1 was significantly diminished after either the GR antagonist RU38486 (4 > 2 = 0.5 microg = vehicle) or a GR antisense oligonucleotide was administered intrathecally twice daily for postoperative day 1-6. Moreover, CCI induced a significant downregulation of nuclear factor kappaB (NF-kappaB) within the ipsilateral spinal cord dorsal horn, which also was attenuated by either RU38486 (4 > 2 = 0.5 microg = vehicle) or a GR antisense oligonucleotide. The immunohistochemical data indicated a pattern of colocalization between GR and EAAC1 as well as GR and NF-kappaB within the spinal cord dorsal horn. Since, NF-kappaB has been shown to regulate the expression of those cellular elements linked to inflammation and tissue injury and its activity can be negatively regulated by GR activation, these results suggest that spinal GR through NF-kappaB may play a significant role in the regulation of EAAC1 expression after peripheral nerve injury, a cellular pathway that may contribute to the development of neuropathic pain behaviors in rats. PMID- 16360275 TI - Presynaptic low- and high-affinity kainate receptors in nociceptive spinal afferents. AB - Presynaptic ionotropic glutamate receptors are increasingly attributed a role in the modulation of sensory input at the first synapse of dorsal root ganglion (DRG) neurons in the spinal dorsal horn. Central terminals of DRG neurons express AMPA and NMDA receptors whose activation modulates the release of glutamate, the main transmitter at these synapses. Previous work, with an antibody that recognizes all low-affinity kainate receptor subunits (GluR5, 6, 7), provided microscopic evidence of presynaptic kainate receptors in unidentified primary afferent terminals in superficial laminae of the spinal dorsal horn (Hwang SJ, Pagliardini S, Rustioni A, Valtschanoff JG. Presynaptic kainate receptors in primary afferents to the superficial laminae of the rat spinal cord. J Comp Neurol 2001; 436: pp. 275-289). We show here that, although all such subunits may be expressed in these terminals, GluR5 is the subunit most readily detectable at presynaptic sites in sections processed for immunocytochemistry. We also show that the high-affinity kainate receptor subunits KA1 and KA2 are expressed in central terminals of DRG neurons and are co-expressed with low-affinity receptor subunits in the same terminals. Quantitative data show that kainate-expressing DRG neurons are about six times more likely to express the P2X(3) subunit of the purinergic receptor than to express substance P. Thus, nociceptive afferents that express presynaptic kainate receptors are predominantly non-peptidergic, suggesting a role for these receptors in the modulation of neuropathic rather than inflammatory pain. PMID- 16360276 TI - Worrying about chronic pain. An examination of worry and problem solving in adults who identify as chronic pain sufferers. AB - Worry is a common feature of chronic pain. The purpose of this study was to examine the extent of worry experienced by adults with chronic pain, and to explore features of problem solving associated with worry and chronic pain. A further purpose was to investigate whether there were differences in worry and problem solving for those presenting at a pain clinic for treatment, compared to those who identified as chronic pain sufferers but who were not presenting for treatment. A final purpose was to examine whether the extent of worry and problem solving attitudes helped to predict the primary clinical features of chronic pain such as intensity, disability and depressive mood. One hundred and eighty five adults with chronic pain provided completed questionnaires assessing worrying, problem solving, pain severity, disability, catastrophic thinking and depressive mood. Analyses showed that worry and problem solving approaches sampled in this study were not abnormal. Furthermore, no differences were found between the clinical and non-clinical sample for worrying and problem solving. In relation to the predictive value of worrying and problem solving, analyses revealed that only worrying had a unique contribution in explaining depressive mood. The results are discussed within a context of a model of worry and chronic pain, in which worry about chronic pain may be functioning to promote awareness of an insoluble problem. PMID- 16360277 TI - The high threshold mechanotransducer: a status report. PMID- 16360280 TI - Atomic force microscopy and confocal laser scanning microscopy on the cytoskeleton of permeabilised and embedded cells. AB - We describe a technical method of cell permeabilisation and embedding to study the organisation and distribution of intracellular proteins with aid of atomic force microscopy and confocal laser scanning microscopy in identical areas. While confocal laser scanning microscopy is useful for the identification of certain proteins subsequent labelling with markers or antibodies, atomic force microscopy allows the observation of macromolecular structures in fixed and living cells. To demonstrate the field of application of this preparatory technique, cells were permeabilised, fixed, and the actin cytoskeleton was stained with phalloidin rhodamine. Confocal laser scanning microscopy was used to show the organisation of these microfilaments, e.g. geodesic dome structures. Thereafter, cells were embedded in Durcupan water-soluble resin, followed by UV-polymerisation of resin at 4 degrees C. This procedure allowed intracellular visualisation of the cell nucleus or cytoskeletal elements by atomic force microscopy, for instance to analyse the globular organisation of actin filaments. Therefore, this method offers a great potential to combine both microscopy techniques in order to understand and interpret intracellular protein relations, for example, the biochemical and morphological interaction of the cytoskeleton. PMID- 16360279 TI - The analgesic effect of low dose focal irradiation in a mouse model of bone cancer is associated with spinal changes in neuro-mediators of nociception. AB - Despite the widespread use of radiotherapy to treat painful bone metastases, the mechanism underlying the analgesic effect of low dose ionizing radiation is unknown. Bone cancer pain is mostly associated with an inflammatory response dominated by local activation of osteoclasts and by astrogliosis in the spinal cord. We determined the effects of a 6 Gy irradiation given focally on osteolytic sarcoma cells inoculated in humeri of mice. Pain behavior was assessed using the rota-rod and the grip force test. Seven days post-irradiation (day 17 post-tumor implantation) the performance of mice markedly improved on the rotarod (non irradiated, 67+/-16s vs irradiated, 223 +/- 22 s; P = 0.0005), and the grip force test (non-irradiated, 34 +/- 4 g vs irradiated, 55 +/- 2 g; P = 0.001). This improvement was similar to the analgesia achieved with 30 mg/kg of the cyclooxygenase (COX) inhibitor ketorolac (Rota-rod, 67 +/- 16 s vs 178 +/- 35 s; P = 0.01: grip force test, 34 +/- 4 g, vs 60 +/- 5 g; P = 0.003). Following irradiation, the tumor mass and the number of osteoclasts did not decrease while the expression of two pro-inflammatory cytokines (monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-alpha) increased. Tumor irradiation led to clear differences in the spinal cord. These include a decrease in glial activity (astrocytes and microglial cells) as well as pain mediators such as dynorphin, COX-2 and chemotactic cytokine receptor (CCR2). We conclude that the analgesic effect of low dose irradiation of bone cancer is associated with the alteration of nociceptive transmission in the central nervous system. PMID- 16360281 TI - Overexpression of gastrin-releasing peptide receptor induced layer disorganization in brain. AB - Gastrin-releasing peptide-preferring and neuromedin B-preferring receptors, members of the bombesin-like peptide receptor subfamily, are reported to regulate proliferation, migration and differentiation. Since they are expressed in developing brain, we postulated that the gastrin-releasing peptide-preferring and neuromedin B-preferring receptors might be involved in normal brain development. Here we examined the effects of the overexpressions of the gastrin-releasing peptide-preferring and neuromedin B-preferring receptors on chick brain development in vivo using a retrovirus. In the overexpressed exogenous gastrin releasing peptide-preferring receptor brain, we found laminar disorganization in the telencephalon, tectum and particularly in the cerebellum with severe atrophy. Processes of the radial glial cells in the telencephalon and optic tectum, as well as the projections of the Bergmann glia in the cerebellum were distorted, which might disturb normal cell migration. Despite the atrophy of the cerebellum, densely-stained proliferating cell nuclear antigen- and phospho-histone H3 positive cells increased in number. Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling-positive cells also increased in the cerebellum, suggesting that the ectopically proliferating cells were subjected to apoptosis. Glial fibrillary acidic protein-positive cells also increased in the hyperpallium accessorium and in the outer layers of the tectum. We also found smaller and spindle-shaped cells which resembled undifferentiated embryonic tumor cells. On the other hand, the layer structures of the neuromedin B-preferring receptors overexpressed brain were well organized and developed, and the size of brain was generally enlarged. These results indicated that although the gastrin-releasing peptide-preferring and neuromedin B-preferring receptors are involved in normal brain development, both receptors contribute and exert their effects differently. PMID- 16360282 TI - Differential response dynamics of corticothalamic glutamatergic synapses in the lateral geniculate nucleus and thalamic reticular nucleus. AB - The corticothalamic feedback pathway provides excitatory synaptic input to both the thalamic reticular nucleus and the lateral geniculate nucleus. We studied excitatory postsynaptic currents elicited from corticothalamic stimulation in the visual sector of the thalamic reticular nucleus and the lateral geniculate nucleus to compare the response of these neurons to stimulation of their common input pathway. Using whole cell patch clamp recordings in ferret thalamic slices, we compared single excitatory postsynaptic current decay kinetics, presynaptic glutamate release dynamics through paired pulse facilitation and responses to corticothalamic train stimulation. We found that single thalamic reticular nucleus excitatory postsynaptic currents were significantly sharper than lateral geniculate nucleus responses. The mean thalamic reticular nucleus excitatory postsynaptic current decay constant (tau) was 4.9+/-0.5 ms, while the mean lateral geniculate nucleus excitatory postsynaptic current tau value was 11.8+/ 0.8 ms. Presynaptic release dynamics as measured by responses to paired stimuli were conserved between the thalamic reticular nucleus and lateral geniculate nucleus. However, facilitating responses to train stimulation were markedly different between nuclei. Lateral geniculate nucleus responses showed proportionately larger facilitation (reaching 842.9 +/- 76.4% of excitatory postsynaptic current 1 amplitude) than thalamic reticular nucleus responses (reaching 223.1 +/- 44.0% of excitatory postsynaptic current 1 amplitude). These data indicate that while the corticothalamic pathway produces excitatory postsynaptic currents in both the thalamic reticular nucleus and lateral geniculate nucleus, other factors uniquely affect the functional integration of the inputs in each nucleus. PMID- 16360283 TI - Peripheral injection of lipopolysaccharide prevents brain recruitment of leukocytes induced by central injection of interleukin-1. AB - I.c.v. injection of interleukin-1beta induces infiltration of leukocytes into the brain. I.p. injection of bacterial endotoxin lipopolysaccharide induces the expression of interleukin-1 in the CNS without causing the entry of leukocytes into the brain. This suggests that during systemic inflammation trafficking of potentially damaging leukocytes into the CNS is inhibited. In this study, we investigated the effects of peripheral injection of lipopolysaccharide on brain leukocyte recruitment induced by i.c.v.-interleukin-1 in mice. I.c.v.-interleukin 1 induced widespread infiltration of leukocytes into the brain 16 h after the injection. Pretreatment with i.p.-lipopolysaccharide 2 h before the i.c.v. interleukin-1 injection completely blocked interleukin-1-induced leukocyte infiltration, whereas i.p.-LPS only attenuated the effect of interleukin-1 if it was given 12 h before i.c.v. interleukin-1 injection. I.p.-lipopolysaccharide given 24 h before i.c.v. interleukin-1 injection did not alter interleukin-1 induced leukocyte infiltration. I.c.v.-interleukin-1 induced expression of p- and e-selectins in brain vasculatures prior to the appearance of leukocytes in the brain parenchyma. Induction of p- and e-selectin was inhibited by the pretreatment of i.p.-lipopolysaccharide 2 h, but not 24 h, before i.c.v. interleukin-1 injection. I.c.v.-interleukin-1-induced leukocyte infiltration was diminished in both e- and p- selectin knockout animals. These results suggest that systemic inflammation actively inhibits recruitment of leukocytes by CNS. Inhibition of the expression of p- and e-selectins is a mechanism by which peripheral inflammation regulate CNS leukocyte recruitment. PMID- 16360284 TI - Agents that act by different mechanisms modulate the activity of protein kinase CbetaII isozyme in the rat spinal cord during peripheral inflammation. AB - Hyperalgesia following unilateral complete Freund's adjuvant-induced inflammation was characterized by paw withdrawal latency to thermal stimulus. Paw withdrawal latencies were significantly shorter on the complete Freund's adjuvant-treated paw than on the contralateral paw of the complete Freund's adjuvant- and the sham treated rats. Total cytosolic protein kinase C activity in the lumbar enlargement was unchanged on the sides of the spinal cord ipsi- and contra-lateral to the inflamed paw. Membrane-associated activities of protein kinase Calpha, protein kinase CbetaI and protein kinase Cgamma did not change significantly on the sides of the cord ipsi- and contra-lateral to the inflammation. However, membrane associated activity of protein kinase CbetaII was increased in the cord section ipsilateral to the inflammation, suggesting that increased translocation/activation of protein kinase CbetaII is related to thermal hyperalgesia. Dextrorphan (an N-methyl-D-aspartate receptor antagonist), L 703,606 (an NK-1 receptor antagonist) and an antisense oligodeoxynucleotide for a selective knockdown of protein kinase Cbeta, reduced complete Freund's adjuvant induced hyperalgesia, and reversed significant changes in the membrane activity of protein kinase CbetaII on the spinal cord section ipsilateral to the inflamed paw. Dextrorphan and protein kinase Cbeta antisense oligodeoxynucleotide were effective in reversing complete Freund's adjuvant-induced increase in the activity of protein kinase CbetaII ipsilateral to the inflammation at all the doses tested, but L-703,606 was effective only at the highest dose. Furthermore, in the presence of inflammatory stimulus, dextrorphan and L-703,606 did not alter the activities of membrane-associated protein kinase Calpha, protein kinase CbetaI, and protein kinase Cgamma in the section of the spinal cord ipsi- and contra-lateral to the inflammation. Protein kinase Cbeta antisense oligodeoxynucleotide had no significant effect on the membrane-associated activities of protein kinase Calpha and protein kinase Cgamma, but decreased the activities of both protein kinase CbetaI and protein kinase CbetaII and the expression of protein kinase Cbeta isozyme in the spinal cord. The data provide evidence that a common molecular event that converges to initiate and maintain hyperalgesia may include the translocation and activation of protein kinase CbetaII in the spinal dorsal horn. PMID- 16360285 TI - Zinc modulates primary afferent fiber-evoked responses of ventral roots in neonatal rat spinal cord in vitro. AB - Zinc ions (Zn(2+)) are known to modulate the functions of a variety of channels, receptors and transporters. We examined the effects of Zn(2+) on the reflex potentials evoked by electrical stimulation and responses to depolarizing agents in the isolated spinal cord of the neonatal rat in vitro. Zn(2+) at low concentrations (0.5-2 microM) inhibited, but at high concentrations (5 and 10 microM) augmented, a slow depolarizing component (slow ventral root potential). Zn(2+) had no effect on fast components (monosynaptic reflex potential; fast polysynaptic reflex potential). Unlike Zn(2+), strychnine (5 microM), a glycine receptor antagonist, and (S),9(R)-(-)-bicuculline methobromide (10 microM), a GABA(A) receptor antagonist, potentiated both fast polysynaptic reflex potential and slow ventral root potential. Zn(2+) (5 microM) did not affect depolarizing responses to glutamate and N-methyl-D-aspartate. Zn(2+) enhanced the substance P evoked depolarization in the absence of tetrodotoxin (0.3 microM) but not in its presence. The dorsal root potential was inhibited by (S),9(R)-(-)-bicuculline methobromide (10 microM) but not by Zn(2+) (5 microM). The Zn(2+)-potentiated slow ventral root potential was inhibited by the N-methyl-D-aspartate receptor antagonists, ketamine (10 microM) and DL-2-amino-5-phosphaonovaleric acid (50 microM) but not by P2X receptor antagonists, pyridoxal-phosphate-6-azophenyl 2',4'-disulphonic acid (30 microM) and 2',3'-O-(2,4,6-trinitrophenyl)ATP (10 microM). Ketamine (10 microM) and DL-2-amino-5-phosphaonovaleric acid (50 microM) almost abolished spontaneous activities increased by Zn(2+). It is concluded that Zn(2+) potentiated slow ventral root potential induced by primary afferent stimulation, which was mediated by the activation of N-methyl-D-aspartate receptors but not by activation of P2X receptors or blockade of glycinergic and GABAergic inhibition. Zn(2+) does not seem to directly affect N-methyl-D aspartate receptors. The release of glutamate from interneurons may play an important role in Zn(2+)-induced potentiation of slow ventral root potential in the spinal cord of the neonatal rat. PMID- 16360286 TI - Melanotan-II: Investigation of the inducer and facilitator effects on penile erection in anaesthetized rat. AB - The effects of melanotan-II, a non-specific agonist of melanocortin receptors, on erection and its possible sites of action were investigated in anesthetized rats. Delivered i.v. (0.1, 0.3 and 1 mg/kg) or within the paraventricular nucleus of the hypothalamus (0.1 and 1 microg), melanotan-II exerted a dose-dependent inducer activity on erection by eliciting erectile events and shortening latency of the first erectile event to occur. Erectile events were of higher amplitude in rats treated with melanotan-II i.t. (0.2 microg) delivered at the L6-S1 level than in animals treated with the vehicle i.t. delivered. Erectile responses elicited by cavernous nerve stimulation were increased after i.v. melanotan-II (1 mg/kg), thereby exerting facilitator effect on erection. In contrast, melanotan II injected within the corpus cavernosum (1 microg) did not display any facilitator activity. To investigate the neural pathways involved in the facilitator effect of melanotan-II, we performed acute spinalization (T8 level) and differential selective nerve transections. Neither spinalization nor bilateral transection of pelvic nerves or dorsal penile nerves impaired facilitator activity of i.v. melanotan-II (1 mg/kg). Conversely, the facilitator effect of melanotan-II was abolished after acute removal of the lumbar paravertebral sympathetic chain. These results lead to the conclusion that central and peripheral melanocortin pathways are recruited by melanotan-II, depending on its route of delivery, to exert both inducer and facilitator activities on erection. PMID- 16360287 TI - The reciprocal connections of endomorphin 1- and endomorphin 2-containing neurons between the hypothalamus and nucleus tractus solitarii in the rat. AB - In the CNS, endomorphin 1- and endomorphin 2-immunoreactive neuronal cell bodies have been principally found both in the hypothalamus and nucleus tractus solitarii. Functionally, the hypothalamus and nucleus tractus solitarii are closely related in many aspects, especially in visceral functions. On the other hand, there are also many endomorphin-immunoreactive fibers and terminals in the two regions. In the present study, to investigate whether endomorphin 1 immunoreactive and endomorphin 2-immunoreactive neurons in the hypothalamus and nucleus tractus solitarii project reciprocally between these two regions, fluorescent retrograde labeling combined with immunofluorescence histochemical staining for endomorphin 1 and endomorphin 2 was used. After injection of Fluoro Gold into the nucleus tractus solitarii of rats, endomorphin 1/Fluoro-Gold or endomorphin 2/Fluoro-Gold double-labeled neuronal cell bodies were predominantly observed in the arcuate nucleus of the hypothalamus, a few of which were also observed in the posterior hypothalamic area and periventricular hypothalamic nucleus. After injection of Fluoro-Gold into the medial zone of hypothalamic tuberal region and the lateral hypothalamic area, respectively, endomorphin 1/Fluoro-Gold or endomorphin 2/Fluoro-Gold double-labeled neuronal cell bodies were found chiefly in the medial, commissural, lateral and gelatinous parts of the nucleus tractus solitarii. These results provide morphological evidence that there exist reciprocal endomorphinergic connections between the hypothalamus and nucleus tractus solitarii. PMID- 16360288 TI - Alcohol expectancies for social facilitation: A short form with decreased bias. AB - The social facilitation scale of the Alcohol Expectancy Questionnaire-Adolescent version predicts subsequent drinking behavior and covaries with other important constructs. An examination of the interval-level measurement properties and potential biases of items could help improve the scale. Responses in a large sample (N=518) of undergraduates confirmed that the items fit a Rasch model for a single-factor, interval scale. Two items were biased when comparing non-drinkers, social drinkers and extreme drinkers. Four additional items showed gender bias. Removal of these items shortened the scale without decreasing correlations with two measures associated with drinking, while the strength with one measure was slightly decreased. This short form of the questionnaire should prove useful in the study of alcohol expectancies for social facilitation without drinking- or gender-related bias. These results also support the utility of Rasch modeling. PMID- 16360289 TI - The role of saturation of fat depots in the pathogenesis of insulin resistance. AB - Insulin resistance is the earliest observable abnormality in individuals who are predisposed to, and who later develop type 2 diabetes mellitus. We hypothesize that saturation of the subcutaneous fat depot is the primary event in the pathophysiology of insulin resistance in the majority of patients and postulate that this seminal event may lead to the development of hypertension, hypertriglyceridemia and depressed HDL levels (i.e., the metabolic syndrome). Our hypothesis has the following clinical implications: (1) differing responses to weight loss may be seen with regards to insulin resistance depending on the size of the fat depot; individuals with small fat depots having to maintain an extremely low body mass to preserve an insulin sensitive phenotype while individuals with a large fat depot may become insulin sensitive even when still clinically obese with some amount of weight loss; (2) peroxisome proliferator activated receptor gamma agonists, such as thiazoledinediones which expand the subcutaneous fat depot, may be especially useful in improving insulin resistance in individuals with small fat depots; (3) expanding alternate storage sites for triglycerides by a variety of techniques, such as resistance training-induced muscle hypertrophy, may also improve insulin resistance; (4) drugs, such as beta 3 adrenergic receptor agonists which promote lipolysis and have been suggested as possible agents in the treatment of obesity may actually increase insulin resistance by releasing free fatty acids into the circulation. Similarly, inhibitors of the beta oxidation of free fatty acids (e.g., carnitine palmitoyl transferase inhibitors) may also actually cause insulin resistance by sparing fat from oxidation and thus worsening fat depot saturation and (5) liposuction, by reducing the size of the subcutaneous fat depot may actually worsen insulin resistance, thus increasing the risk of type 2 diabetes mellitus. PMID- 16360290 TI - Evolution of metabolic networks by gain and loss of enzymatic reaction in eukaryotes. AB - The metabolic network is composed of enzymatic reactions (ERs) in which one or more enzymes catalyze the reaction of pertinent substrates. Since metabolism is a basal system for maintaining life of all organisms, any change in the metabolic networks must greatly affect organismic evolution. The aim of this study is to examine how often gains and losses of ER have occurred during the evolution of metabolic networks in eukaryotes and how these evolutionary events have affected phenotypic traits of organisms. In this study, we conducted comparative studies of 751 ERs in the metabolic networks of 6 eukaryotic species whose complete genome sequences were determined. As a result, we found that a total of 804 gains and losses of ERs had occurred in the evolutionary diversification of metabolic networks in different lineages. Moreover, the vertebrate lineage, after the separation from Drosophila melanogaster, showed a remarkable increase in the number of ER gains compared with ER losses. In particular, 41% of the ER gains were predominantly involved with lipids and complex lipid metabolism. Because some products of these two metabolisms function as hormones, we concluded that the ER gain of these two metabolisms accelerated the development of hormonal signal transduction for the elaborate regulation of physiological systems during vertebrate evolution. PMID- 16360291 TI - Characterization, organization and expression of AmphiLysC, an acidic c-type lysozyme gene in amphioxus Branchiostoma belcheri tsingtauense. AB - The study on lysozymes remains open in amphioxus, a cephalochordate. Here we show the existence of c-type lysozyme gene (AmphiLysC) in amphioxus, first such data in the basal chordates including urochordate and cephalochordate. This is in contrast to the absence of c-type lysozyme genes in urochordate. It is found that there exist two copies of c-type lysozyme genes in amphioxus genome, and their gene organization is similar to vertebrate c-type lysozyme genes with respect to the number and the size of both exons and introns. AmphiLysC possesses main features characteristic of the digestive c-type lysozyme such as lower number of basic amino acids (low pI values) and pH-optimum in acidic range. Moreover, AmphiLysC is predominantly expressed in the gut. These indicate that AmphiLysC is possibly a digestive c-type enzyme. However, the ubiquitous expression of AmphiLysC in non-digestive tissues such as ovaries, testes, notochord, gill and muscle suggests that it may also play a non-digestive role like antibacterial activity. It is highly likely that AmphiLysC is an enzyme with a combined function of digestion and bacteriolysis. PMID- 16360292 TI - Ontogeny of the human central nervous system: what is happening when? AB - The present paper reviews current data on the structural development of the human nervous system. Focus is on the timing of ontogenetic events in the telencephalon. Neuronal proliferation and migration especially occur during the first half of gestation; the second half of gestation is the period of the existence of the functionally important transient structure 'subplate' and the major period of glial cell proliferation and programmed cell death. Axon and dendrite sprouting and synapse formation bloom during the last trimester of gestation and the first postnatal year. Major part of telencephalic myelination occurs during the first year after birth. Many developmental processes, such as myelination, synapse formation and synapse elimination continue throughout childhood and adolescence. Evidence is emerging that the peak of synapse elimination occurs between puberty and the onset of adulthood. Neurotransmitter systems are present from early foetal life onwards and their pre- and perinatal development is characterized by periods of transient overexpression. The latter is for instance true for the acetylcholinergic, catecholaminergic and glutamate systems. Thus, the development of the human brain is characterized by a protracted, neatly orchestrated chain of specific ontogenetic events. The continuous changes of the nervous system have consequences for vulnerability to adverse conditions, for diagnostics and for physiotherapeutical intervention. PMID- 16360293 TI - Subnormal head circumference in very low birth weight children: neonatal correlates and school-age consequences. AB - BACKGROUND: Subnormal head circumference (HC) has been associated with poor neurologic and developmental outcomes. AIM: To examine the correlates and consequences of subnormal HC in a cohort of school age, very low birth weight (VLBW, <1,500 g) children. STUDY DESIGN AND OUTCOME MEASURES: We examined developmental outcomes at a mean age of 6.8 years in a cohort of 128 VLBW children born from 1982-1986 and 58 normal birth weight controls. The VLBW cohort included a regional sample of <750 g birth weight children and matched children with 750-1,499 g birth weight. Outcomes included an IQ equivalent, neuropsychological skills, academic achievement, adaptive behavior, and attention problems. HC was defined along a continuum and as subnormal vs. normal. Linear and logistic regressions were employed to determine the effects of HC on the outcomes after controlling for confounding variables. RESULTS: Thirty one VLBW children (24%) had subnormal HC vs. none of the controls. The VLBW children with subnormal HC differed significantly from the normal HC group in birth weight (748 g vs. 977 g, p<.001), SGA status (52% vs. 27%, p<.05), high neonatal risk (57% vs. 29%, p<.05), and neurosensory impairment (23% vs. 8%, p<.05). Even after taking these risk factors into account, subnormal HC was associated with poorer IQ equivalent, perceptual motor skills, academic achievement, and adaptive behavior. Results were similar after excluding the children with neurosensory impairment. CONCLUSIONS: Subnormal HC is associated with adverse developmental outcomes in VLBW children, independent of other risk factors. Interventions to improve antenatal and postnatal growth may contribute to better school-age outcomes. PMID- 16360294 TI - Developing a SNP panel for forensic identification of individuals. AB - Single nucleotide polymorphisms (SNPs) are likely in the near future to have a fundamental role in forensics in both human identification and description. However, considerable research is necessary to establish adequate scientific foundations for these applications. In the case of identification, because allele frequencies can vary greatly among populations, the population genetics of match probabilities is a critical issue. Some SNPs, however, show little allele frequency variation among populations while remaining highly informative. We describe here both an efficient strategy for identifying and characterizing such SNPs, and test that strategy on a broad representation of world populations. Markers with high heterozygosity and little frequency variation among African American, European American, and East Asian populations are selected for additional screening on seven populations that provide a sampling of genetic variation from the world's major geographical regions. Those with little allele frequency variation on the seven populations are then screened on a total of 40 populations ( approximately 2100 individuals) and the most promising retained. The preliminary panel of 19 SNPs, from an initial selection of 195 SNPs, gives an average match probability of <10(-7) in most of 40 populations studied and no greater than 10(-6) in the most isolated, inbred populations. Expansion of this panel to approximately 50 comparable SNPs should give match probabilities of about 10(-15) with a small global range. PMID- 16360295 TI - Effects of colostrum feeding and glucocorticoid administration on insulin dependent glucose metabolism in neonatal calves. AB - Colostrum feeding and glucocorticoid administration affect glucose metabolism and insulin release in calves. We have tested the hypothesis that dexamethasone as well as colostrum feeding influence insulin-dependent glucose metabolism in neonatal calves using the euglycemic-hyperinsulinemic clamp technique. Newborn calves were fed either colostrum or a milk-based formula (n=14 per group) and in each feeding group, half of the calves were treated with dexamethasone (30 microg/[kg body weight per day]). Preprandial blood samples were taken on days 1, 2, and 4. On day 5, insulin was infused for 3h and plasma glucose concentrations were kept at 5 mmol/L+/-10%. Clamps were combined with [(13)C]-bicarbonate and [6,6-(2)H]-glucose infusions for 5.5h (i.e., from -150 to 180 min, relative to insulin infusion) to determine glucose turnover, glucose appearance rate (Ra), endogenous glucose production (eGP), and gluconeogenesis before and at the end of the clamp. After the clamp liver biopsies were taken to measure mRNA levels of phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate carboxylase (PC). Dexamethasone increased plasma glucose, insulin, and glucagon concentrations in the pre-clamp period thus necessitating a reduction in the rate of glucose infusion to maintain euglycemia during the clamp. Glucose turnover and Ra increased during the clamp and were lower at the end of the clamp in dexamethasone-treated calves. Dexamethasone treatment did not affect basal gluconeogenesis or eGP. At the end of the clamp, dexamethasone reduced eGP and PC mRNA levels, whereas mitochondrial PEPCK mRNA levels increased. In conclusion, insulin increased glucose turnover and dexamethasone impaired insulin-dependent glucose metabolism, and this was independent of different feeding. PMID- 16360296 TI - WWOX--the FRA16D cancer gene: expression correlation with breast cancer progression and prognosis. AB - AIMS: WWOX is a tumour suppressor gene involved in various tumours including breast cancer. High chromosomal abnormalities in a genomic region spanned by WWOX are associated with the fact that this gene covers approximately 1 million base pairs of the second most affected among common chromosomal fragile sites FRA16D. We evaluated WWOX expression levels in breast cancer samples in association with diagnostics-prognostics markers. METHODS: We performed quantitative real-time RT PCR to analyse levels of expression of WWOX in 132 cases of breast cancer. We evaluated the relationship between WWOX mRNA levels, clinico-pathological factors, expression of aberrant WWOXDelta6-8 mRNA and other cancer related genes. RESULTS: Expression of WWOX was higher in patients younger than 50 years old, in ER and PR positive tumours vs negative for those receptors and tumours without lymph node metastasis vs LN+. WWOX mRNA levels were also higher in tumours with higher apoptotic index (Bcl2/Bax ratio). Negative associations were found between WWOX expression and cytokeratins 5/6 and 17 (P<0.05). High level expression of WWOX was also associated with better disease free survival. Presence of WWOXDelta6-8 transcripts were accompanied with lower WWOX wild type mRNA level. CONCLUSIONS: Reduced WWOX expression commonly observed in various neoplasias in cases of breast cancer is associated with markers of bad prognosis. Our findings reveal additional evidence that WWOX may be involved in steroid (estrogens) metabolism and signaling pathways. WWOX can be considered as a new target for gene therapy development due to the association of high WWOX expression with improved disease free survival. PMID- 16360297 TI - [Intensive care management of HELLP syndrome in Dakar]. AB - OBJECTIVE: To evaluate the intensive management of HELLP syndrome in a intensive care unit in African setting. STUDY DESIGN: Descriptive and analytical retrospective study. PATIENTS AND METHODS: All patients hospitalized between June 1998 and June 2004 for HELLP syndrome were included in the study. Following parameters were studied: age, parity and gestity, term of pregnancy, delay before admission in ICU, data clinical, biological parameters, medical treatment, obstetrical treatment, maternal complications, maternal and foetal prognosis. RESULTS: Twenty patients of average age 26.3 years with seven primigeste and 13 pauci and miltigeste were admitted in ICU during the period of study for HELLP syndrome. The average time of admission was 1.35 days; 13 patients had HELLP syndrome in ante partum and seven patients in postpartum. Serious complications were noted at admission in the majority of patients with oligoanuric renal insufficiency in 11 cases, eclampsia in eight cases and intravascular coagulation disseminated in four cases. The mode of delivery in patients for HELLP syndrome occurring in ante partum was the Caesarean section under general anaesthesia in five cases and vaginal delivery in eight cases. All the patients for HELLP syndrome in the postpartum had been confined by low way. The maternal evolution was favourable in 13 cases. Seven patients of which five with HELLP syndrome who occurred in ante partum had died. The infant mortality was noted in eight cases primarily in cases of HELLP syndrome which occurred in ante partum. PMID- 16360298 TI - [Dyskalemia and head injury]. PMID- 16360299 TI - [Case report: a bacteremic pasteurellosis]. AB - Bacteremic pasteurellosis is an uncommon form of Pasteurella multocida infection, usually involved in local infections. This systemic infection often occurs in immuno-compromised patient such as cirrhotic or alcoholic patients, with a high mortality rate (up to 60%). Septic shock may occur and neurological disorders or coma are frequent. We report such a case. Treatment associated local care, antibiotics (beta-lactam antibiotics plus fluoroquinolone) during 14 days and resuscitation of septic shock. Owing these therapies, septic shock was successfully treated without complications. PMID- 16360301 TI - "Hot potato voice" in peritonsillitis: a misnomer. AB - The "hot potato voice" is widely recognized as a symptom of peritonsillar cellulitis or abscess; yet there have been no studies assessing the resonance characteristics of the vocal tract in peritonsillitis. Analysis was undertaken of formant frequencies in the articulation of the vowels /i:/. /a:/ and /u:/ in six subjects with peritonsillitis and compared with articulation once the peritonsillitis had settled. Significant variation was found in F1 when articulating /i:/ and in F2 when articulating /a:/, which are explainable by dyskinesis of the peritonsillar musculature. These findings were compared with six subjects articulating the same vowels with and without a hot potato in their mouth. Variation was found in both F1 and F2 when articulating /i:/, which can be related to interference of the potato with movement of the anterior tongue. The changes in the vocal tract differ in these two cases and the title "hot potato voice" in peritonsillitis is a misnomer. PMID- 16360300 TI - Effects of diesel exhaust particles on human alveolar macrophage ability to secrete inflammatory mediators in response to lipopolysaccharide. AB - Ambient particulate matter (PM) has been shown to be associated with mortality and morbidity. Diesel exhaust particles (DEP) contribute to ambient PM. Alveolar macrophages (AM) are important targets for PM effects in the lung. The effects of DEP exposure on human AM response to lipopolysachharide (LPS; from gram-negative bacteria) challenge in vitro were determined by monitoring the production of interleukin 8 (IL-8), tumor necrosis factor-alpha (TNF-alpha) and prostaglandin E(2) (PGE(2)). The roles of organic compounds and carbonaceous core of DEP in response to LPS were evaluated by comparing the DEPs effect to that of carbon black (CB), a carbonaceous particle with few adsorbed organic compounds. AMs were exposed in vitro to Standard Reference Material (SRM) DEP 2975, SRM DEP 1650, SRM 1975 (a dichloromethane extract of SRM DEP 2975) and CB particles for 24 h. DEPs induced a decreased secretion of IL-8, TNF-alpha and PGE(2) in response to a subsequent LPS stimulation. DEPs also show suppressive effect on the release of inflammatory mediators when stimulated with lipoteichoic acid, a product of gram positive bacteria. In summary, in vitro exposure of human AM to DEPs significantly suppress AM responsiveness to gram-negative and positive bacterial products, which may be a contributing factor to the impairment of pulmonary defense. PMID- 16360302 TI - The prevalence of childhood dysphonia: a cross-sectional study. AB - There is only very limited information on the prevalence of voice disorders, particularly for the pediatric population. This study examined the prevalence of dysphonia in a large cohort of children (n = 7389) at 8 years of age. Data were collected within a large prospective epidemiological study and included a formal assessment by one of five research speech and language therapists as well as a parental report of their child's voice. Common risk factors that were also analyzed included sex, sibling numbers, asthma, regular conductive hearing loss, and frequent upper respiratory infection. The research clinicians identified a dysphonia prevalence of 6% compared with a parental report of 11%. Both measures suggested a significant risk of dysphonia for children with older siblings. Other measures were not in agreement between clinician and parental reports. The clinician judgments also suggested significant risk factors for sex (male) but not for any common respiratory or otolaryngological conditions that were analyzed. Parental report suggested significant risk factors with respect to asthma and tonsillectomy. These results are discussed in detail. PMID- 16360303 TI - Laryngeal biomechanics in Middle Eastern singing. AB - We would like to describe the muscle tension patterns observed in Middle Eastern singing and correlate these findings with demographic data. MATERIAL: A total of 42 candidates were included in this study. The medical charts and video records of their fiberoptic nasopharyngeal laryngoscopy were reviewed. Demographic information included age, sex, history of smoking, history of reflux, history of voice overuse/abuse, status in singing (professional vs. amateur), and style of singing (classical vs pop). Each frame was analyzed for each of the four muscle tension patterns (MTPs): MTP I, MTP II, MTP III, and MTP IV. STATISTICAL METHOD: Nonparametric Mann-Whitney U test was conducted. RESULTS: Twenty-six subjects were professional singers and half sang classical singing. The mean MTP total score for all subjects was 35.58 +/- 21.98% (+/- standard deviation). The highest muscle tension score was obtained for type III pattern followed by type II. There were no statistical differences in the mean MTP total, MTP II, and MTP III scores among the professional singers versus the amateurs. Similar results were obtained when MTP scores were compared by style of singing. CONCLUSION: The vocal technique and status of singing did not seem to affect the muscle tension score. PMID- 16360304 TI - Macrognathia secondary to dialysis-related renal osteodystrophy treated successfully by parathyroidectomy. AB - Renal osteodystrophy (ROD) is one of the most common complications affecting patients with chronic renal failure both before and after the initiation of maintenance dialysis, but macrognathia secondary to ROD is rare. Usually, enlarged jaws due to ROD do not return to their normal contours after the treatment of hyperparathyroidism. To the authors' knowledge, this article describes the second case of macrognathia secondary to dialysis-related ROD treated successfully by parathyroidectomy. Immunohistochemical study of the maxilla confirmed that parathyroidectomy could stop maladaptive parathyroid hormone stimulation, which leads not only to the formation of osteoblastic progenitors that become fibroblast-like cells but also to osteoclast formation. PMID- 16360305 TI - Validation of a physical activity assessment tool for individuals with schizophrenia. AB - OBJECTIVE: Increasing physical activity must be one component of lifestyle interventions designed to prevent or treat obesity in schizophrenia and there is now a need to develop low cost, practical and accurate measures of physical activity in this population to identify the prevalence of physical (in)activity and to assess the effectiveness of physical activity interventions. The objective of this study was to provide preliminary validation of the Short-Form International Physical Activity Questionnaire (IPAQ), a measurement tool that could prove useful for both clinicians and researchers in the field. METHOD: Reliability and validity data were collected from a sample of 35 outpatients with a DSM-IV diagnosis of schizophrenia. Test-retest repeatability was assessed within the same week and criterion validity was assessed against an RT3 accelerometer. Spearman's correlation coefficients are reported based on the total reported physical activity (minutes) and estimated energy expenditure. RESULT: We found a correlation coefficient of 0.68 for reliability and 0.37 for criterion validity based on total reported minutes of physical activity. There was a nonsignificant correlation (0.30; p>0.05) between the RT3 data and estimated energy expenditure derived from the IPAQ. CONCLUSION: Although not without limitations, the Short-Form IPAQ, when used with individuals with schizophrenia, exhibits measurement properties that are comparable to those reported in the general population and can be considered as a surveillance tool to assess levels of physical activity. PMID- 16360306 TI - Temperament, character and symptoms of anxiety and depression in the general population. AB - Few studies have investigated the relationship of temperament and character, as conceptualized in the Temperament and Character Inventory-Revised (TCI-R), to symptoms of depression and anxiety in the general population. In this study a random sample of subjects (20 to 70 years), in two Finnish cities, were surveyed with the TCI-R, Beck Depression and Anxiety Inventories, plus questions related to diagnosed lifetime mental disorders, health care use for psychiatric reasons during the past 12 months, and history of mental disorders in first-degree relatives. Altogether 347 subjects (38.6%) responded. Of the TCI-R dimensions, Harm Avoidance correlated with symptoms of depression (r(s)=0.555, p<0.001), anxiety (r(s)=0.560, p<0.001), self-reported lifetime mental disorder (r(s)=0.272, p<0.001), health care use for psychiatric reason during the past 12 months (r(s)=0.241, p<0.001) and family history of mental disorder (r(s)=0.202, p<0.001). Self-directedness correlated negatively with symptoms of depression (r(s)=-0.495, p<0.001), anxiety (r(s)=-0.458, p<0.001), lifetime mental disorder (r(s)=0.225, p<0.001) and health care use (r(s)=-0.135, p=0.013). Overall, Harm Avoidance and Self-directedness seem to associate moderately with depressive and anxiety symptoms, and somewhat predict self-reported use of health services for psychiatric reasons, and lifetime mental disorder. High harm avoidance may associate with a family history of mental disorder. PMID- 16360307 TI - Self-help with minimal therapist contact for obsessive-compulsive disorder: a review. AB - Though there are effective psychological and drug treatments for obsessive compulsive disorder (OCD), many patients remain inadequately treated or untreated. Making effective self-treatment guidance available may increase the number of patients being helped. In this review, database and manual literature searches were performed of case studies, open and randomised controlled trials (RCTs) of bibliotherapy, self-help groups, telecare and computer-aided self-help for OCD. We found no RCTs of bibliotherapy or self-help groups for OCD. Three open studies showed the efficacy of brief exposure and ritual prevention (ERP) instructions delivered by a live therapist by phone. A vicarious ERP computer program was effective in a small open study. Fully interactive computer-aided self-help by ERP for OCD was efficacious in two open studies and a large multicentre RCT, and in a small RCT compliance and outcome with that program was enhanced by brief scheduled support from a clinician. Although more research is needed, self-help approaches have the potential to help many more patients who would otherwise remain inadequately treated or untreated. Their dissemination could save resources used by health care providers. We propose a stepped care model for the treatment of OCD. PMID- 16360309 TI - Cigarette smoking in long-term schizophrenia. AB - PURPOSE: Cigarette smoking is a great health problem and prevalent among subjects with schizophrenia. Our aim was to investigate the prevalence and associations of cigarette smoking in patients with long-term schizophrenia. METHODS: Seven hundred and sixty schizophrenia patients were interviewed and their cigarette smoking was recorded. RESULTS: Smoking was more prevalent men than in women patients. In logistic regression analysis, male gender, duration of illness (DUI) from 10 to 19 years, being divorced or separated, lower education and high daily doses of neuroleptics (DDN) associated significantly with regular smoking. Heavy smoking associated, in men, with hospital treatment. CONCLUSIONS: In schizophrenia patients, smoking is associated with long DUI, high DDN and institutional care. Interventions for cessation and/or reduction of cigarette smoking should be a part of the treatment for patients with schizophrenia. PMID- 16360308 TI - Neurological soft signs in adolescents with first episode psychosis. AB - The purpose of this study is to determine the decrease of neurological soft signs (NSS) during adolescence and to compare this evolutionary process in two groups of adolescents with first episode psychosis: a) schizophrenia and b) non schizophrenia patients. The structured neurological evaluation scale (NES) was administered to 24 adolescents with first episode psychosis. The number of NSS, the total and subscales scores were correlated with age in patients and in 39 healthy controls. Adolescents with first-episode psychosis had a higher prevalence of NSS than healthy controls; the schizophrenia patients (N=9) scored higher than non-schizophrenia patients (N=15). The number of NSS, total score and the scores on three of the four NES subscales correlated inversely with age in the healthy control group. No correlation was found for the schizophrenia group. For the non-schizophrenia group, a significant negative correlation was found only in one subscale. The decrease of NSS during adolescence in the healthy population but not in the patient groups with psychosis may be an indicator of a disturbance of brain processes that occurs during development. We did not find a clear pattern of NSS that distinguished schizophrenia from other psychoses. PMID- 16360310 TI - Adjunctive topiramate in ultradian cycling bipolar disorder: case report with 3 year follow-up. AB - A patient with a treatment-refractory bipolar disorder with ultradian cycling responded to adjunctive topiramate. Response was maintained during 3-year follow up. PMID- 16360311 TI - Randomised clinical trial comparing oral versus depot formulations of zuclopenthixol in patients with schizophrenia and previous violence. AB - PURPOSE: The aim of this longitudinal study was to determine whether the depot formulation of an antipsychotic reduces violence in outpatients with schizophrenia as compared to oral administration of the same antipsychotic. METHODS: Forty-six previously violent patients with schizophrenia were randomised to receive treatment with oral or depot zuclopenthixol for 1 year. Clinicians interviewed patients at baseline and every month thereafter to assess treatment adherence. An interviewer blinded to treatment assignments interviewed an informant about any violent behaviour during the previous month. RESULTS: Violence during the follow-up year was inversely proportional to treatment adherence, better compliance, and greater reduction of positive symptoms. Lower frequency of violent acts was observed in the depot group. The level of insight at baseline was not significantly associated with violence recidivism. Regardless of route of administration, treatment non-adherence was the best predictor of violence. CONCLUSIONS: Some patients with schizophrenia and prior violent behaviour may benefit from the depot formulation of antipsychotic medication. PMID- 16360312 TI - AgRP(83-132) and SHU9119 differently affect activity-based anorexia. AB - Activity-based anorexia (ABA) mimics starvation and hyperactivity of anorexia nervosa patients in rats. Activation of the melanocortin (MC) system leads to hypophagia and increased energy expenditure in ad libitum fed rats. Therefore, activation of the MC system might underlie the development and propagation of ABA. Pro-opiomelanocortin (POMC) gene expression is normally decreased during negative energy balance. Strikingly, we found a transient up-regulation of POMC mRNA levels in the arcuate nucleus during the development of ABA, indicating a hyperactive MC system. However, wheel running and food intake were not influenced by treating ABA rats with the competitive antagonist SHU9119. This suggests that agonism of MC receptors by endogenous alpha-melanocyte-stimulating hormone (alpha MSH) levels does not underlie ABA. Instead, treatment with the inverse agonist AgRP(83-132) did ameliorate signs of ABA. This implies that modulation of constitutive MC receptor activity rather than antagonizing putative alpha-MSH release contributes to the development and propagation of ABA. PMID- 16360313 TI - Estimation of odor emission rate from landfill areas using the sniffing team method. AB - The monitoring of the odor annoyance generated by a landfill area is difficult, since it is a multi-area-sources problem, with a discontinuous odor emission. This paper proposes an adaptation of the method of sniffing team campaigns to the particular case of fresh waste odors. The method is based on the field determination of odor perception points, followed by data processing with a bi Gaussian-type model, adapted to handle the odors. In a first step, field observers delineate the region in which odor impact is experienced and then the emission rate is manipulated in a dispersion model until the predicted size of the impact zone matches that observed in the field. In a second step the adjusted emission rate is entered into the model to calculate the percentiles corresponding to the average annoyance zone. The originality of the proposed method is the introduction of all observation points and of all recorded meteorological data into the model. The paper discusses the method limitations and the errors induced on the results, i.e. the odor emission rate and the percentile lines (or iso-concentration lines) which are used to describe the odor concentrations on a map of the surroundings of the plant. The proposed method proves to be reliable for diffuse sources, such as landfill areas. The obtained results are coherent with other results found in the literature with other techniques. PMID- 16360314 TI - Qualitative study of how women define and use information about breast symptoms and diagnostic tests. AB - Unstructured interviews were conducted with 14 women with recent experience of diagnostic testing to explore the definition and role of information in women's experiences of undergoing diagnostic breast tests. Analysis showed that women see information as more than what they are told by healthcare professionals. Information also comes from significant others, administration and reception staff, media, comparison with previous breast symptoms, silences, gestures and behavior of healthcare professionals and significant others, and the number of tests they undergo. Information is about more than understanding. It can provide support and control, reduce anxiety, facilitate question asking and promote women's trust and confidence in themselves and their healthcare professionals. Healthcare professionals need to be aware that women define information more broadly than what they are told, and of the importance women attach to this other information. They also need to be aware that information serves more purposes than helping women understand the diagnostic testing process and their result. PMID- 16360316 TI - Clarification note in reference to "Scanning probe microscopy examination of the surface properties of keratin fibres", by J.M. Maxwell and M.G. Huson. Micron 36 (2005) 127-136. PMID- 16360315 TI - Rapid PIKK-dependent release of Chk1 from chromatin promotes the DNA-damage checkpoint response. AB - BACKGROUND: Checkpoint signaling pathways are of crucial importance for the maintenance of genomic integrity. Within these pathways, the effector kinase Chk1 plays a central role in mediating cell-cycle arrest in response to DNA damage, and it does so by phosphorylating key cell-cycle regulators. RESULTS: By investigating the subcellular distribution of Chk1 by cell fractionation, we observed that around 20% of it localizes to chromatin during all phases of the cell cycle. Furthermore, we found that in response to DNA damage, Chk1 rapidly dissociates from the chromatin. Significantly, we observed a tight correlation between DNA-damage-induced Chk1 phosphorylation and chromatin dissociation, suggesting that phosphorylated Chk1 does not stably associate with chromatin. Consistent with these events being triggered by active checkpoint signaling, inhibition of the DNA-damage-activated kinases ATR and ATM, or siRNA-mediated downregulation of the DNA-damage mediator proteins Claspin and TopBP1, impaired DNA-damage-induced dissociation of Chk1 from chromatin. Finally, we established that Chk1 phosphorylation occurs at localized sites of DNA damage and that constitutive immobilization of Chk1 on chromatin results in a defective DNA damage-induced checkpoint arrest. CONCLUSIONS: Chromatin association and dissociation appears to be important for proper Chk1 regulation. We propose that in response to DNA damage, PIKK-dependent checkpoint signaling leads to phosphorylation of chromatin-bound Chk1, resulting in its rapid release from chromatin and facilitating the transmission of DNA-damage signals to downstream targets, thereby promoting efficient cell-cycle arrest. PMID- 16360317 TI - Monocyte recruitment and foam cell formation in atherosclerosis. AB - Atherosclerosis is a chronic immune-inflammatory disease in which the interactions of monocytes with activated endothelium are crucial events leading to atherosclerotic alteration of the arterial intima. In early atherosclerosis, monocytes migrate into the subendothelial layer of the intima where they differentiate into macrophages or dendritic cells. In the subendothelial space enriched with atherogenic lipoproteins, most macrophages transform into foam cells. Foam cells aggregate to form the atheromatous core and as this process progresses, the atheromatous centres of plaques become necrotic, consisting of lipids, cholesterol crystals and cell debris. This review highlights some aspects of monocyte recruitment and foam cell formation in atherosclerosis. PMID- 16360318 TI - A STEM/EELS method for mapping iron valence ratios in oxide minerals. AB - The valence state of iron in minerals has useful applications in the geosciences for estimating redox conditions during mineral formation or re-equilibration. STEM/EELS techniques offer the advantage over other methods of being able to measure Fe valence with very high spatial resolution across mineral grains and grain boundaries. We have modified an EELS method for point analyses of iron valence ratios (Fe(3+)/SigmaFe) making it possible to generate line scans and maps of Fe valence. We demonstrate this method with measurements at an interface between iron-bearing oxides in a finely intergrown sample of magnetite and ilmenite. The STEM/EELS method is based on a calibrated relationship between Fe(3+)/SigmaFe and the relative intensities of the Fe L(3) and L(2) white lines in core energy-loss spectra for oxide and silicate minerals. Our method overcomes problems of energy drift in spectrum images by aligning energy-loss edges at a fixed energy position prior to background removal. An automated routine for batch processing of core loss spectra, including additional background removal and calculation of Fe L(3)/L(2) intensity ratios, allows for rapid Fe(3+)/SigmaFe determinations of multiple point analyses or spectrum images and the preparation of Fe valence maps, with an analytical error of +/-0.05 to +/-0.09 in the Fe(3+)/SigmaFe measurements. PMID- 16360319 TI - Spider's microstructure for sensing. AB - The spider is well known for sensing the movements of air and preys. Bionics of the spider based on this principle is being paid great attention by many researchers. Here, this paper presents some detailed organs of the spider to make an attempt to clarify the sensing mechanism of the spider from the point view of physical structure by scanning electron microscopy. And behavior characteristics concerning sensing action are observed by optical microscopy. Compared with structures, some novel features of sense movements in micro- and nano-scale size and corresponding possible models are presented. At the same time, simple structure analysis is made to explain and prove this hypothesis. PMID- 16360320 TI - A graphical model for estimating stimulus-evoked brain responses from magnetoencephalography data with large background brain activity. AB - This paper formulates a novel probabilistic graphical model for noisy stimulus evoked MEG and EEG sensor data obtained in the presence of large background brain activity. The model describes the observed data in terms of unobserved evoked and background factors with additive sensor noise. We present an expectation maximization (EM) algorithm that estimates the model parameters from data. Using the model, the algorithm cleans the stimulus-evoked data by removing interference from background factors and noise artifacts and separates those data into contributions from independent factors. We demonstrate on real and simulated data that the algorithm outperforms benchmark methods for denoising and separation. We also show that the algorithm improves the performance of localization with beamforming algorithms. PMID- 16360321 TI - Evidence for a direct association between cortical atrophy and cognitive impairment in relapsing-remitting MS. AB - Cognitive deficits affecting memory, attention and speed of information processing are common in multiple sclerosis (MS). The mechanisms of cognitive impairment remain unclear. Here, we examined the association between neuropsychological test performance and brain atrophy in a group of mildly disabled patients with relapsing-remitting MS. We applied voxel-based morphometry (SPM2) to investigate the distribution of brain atrophy in relation to cognitive performance. Patients had lower scores than control subjects on tests of memory and executive function, including the PASAT, Digit Span Backward and a test of short-term verbal memory (Memo). Among patients, but not healthy controls, performance on the PASAT, a comprehensive measure of cognitive function and reference task for the cognitive evaluation of MS-patients, correlated with global grey matter volume as well as with grey matter volume in regions associated with working memory and executive function, including bilateral prefrontal cortex, precentral gyrus and superior parietal cortex as well as right cerebellum. Compared to healthy subjects, patients showed a volume reduction in left temporal and prefrontal cortex, recently identified as areas predominantly affected by diffuse brain atrophy in MS. A comparison of low performers in the patient group with their matched control subjects showed more extensive and bilateral temporal and frontal volume reductions as well as bilateral parietal volume loss, compatible with the progression of atrophy found in more advanced MS patients. These findings indicate that MS-related deficits in cognition are closely associated with cortical atrophy. PMID- 16360322 TI - Molecular phylogenetics of blackflies of the Simulium damnosum complex and cytophylogenetic implications. AB - The molecular phylogenetics of the Simulium damnosum complex, including vectors of the human parasite Onchocerca volvulus, from various parts of Africa was studied and compared with results of cytogenetic analyses. The sequence data of mitochondrial 16s and nuclear ITS2 rDNA revealed that the complex comprises two main clades, roughly covering the more easterly and westerly African taxa, respectively. However, striking inconsistencies in the tree topologies turned up between the DNA fragments regarding the position of certain subcomplexes and species. The cytophylogenetic relationships are better reflected in the ITS2 tree where Simulium pandanophilum and Simulium mengense constitute a basal, Central African clade of the entire complex and are therefore suggested to be the chromosomal roots too. Further divisions and the corresponding biogeographic interpretations are discussed. Several species and cytoforms are placed within the system for the first time. The phylogenetic relationships within the complex do hardly correlate with host preferences or other behavioral and ecological characteristics. PMID- 16360323 TI - Rolling circle amplification of metazoan mitochondrial genomes. PMID- 16360324 TI - On the osmotically unresponsive water compartment in cells. AB - Differences in colligative properties (freezing point, boiling point, vapor pressure and osmotic behavior) between water in living cells and pure bulk water were investigated by re-evaluating reports of the osmotic behavior of mammalian cells. In five different animal cells, osmotically unresponsive water (OUW) values ranged from 1.1 to 2.2 g per g dry mass. Detailed analysis of human red blood cell (RBC) data indicates a major role for hemoglobin OUW-values, aggregation and packing in cell volume regulation that can be explained for the first time in relevant molecular terms. PMID- 16360325 TI - Repercussions of castration and vasectomy on the ductal system of the rat ventral prostate. AB - Diseases, such as cancer and benign prostatic hyperplasia, are related to disruption of the mechanism regulating the balance between cell proliferation and apoptosis in prostatic cells. Since castration and vasectomy might alter that balance, this study evaluates the cell proliferation, apoptosis and height of the secretory epithelium of the ventral-prostate ductal system post-castration and vasectomy. Immunohistochemical (PCNA and Ki67), cytochemical (Fuelgen reaction) and morphometric investigation have been carried out. Cell proliferation indices decreased significantly in both regions of the ventral-prostate ductal system after castration compared to the sham-operated group. The apoptotic index increased significantly after 48 h, declining 7 days post-castration. The cell proliferation indices did not differ after 48 h significantly; however, they increased 7 days post-vasectomy in both regions. The apoptotic index did not differ significantly in either time post-vasectomy. Castration caused an imbalance in favor of apoptosis, whereas vasectomy caused an imbalance in favor of cell proliferation. PMID- 16360326 TI - Temporary access and central venous catheters. AB - The use of central venous catheters for temporary vascular access is a vital part of modern medicine and has an important role in the management of patients with renal failure. Attention to detail when addressing issues relating to temporary venous access, will pay dividends, with significant reductions in morbidity and mortality in both the short and long term. PMID- 16360327 TI - A preliminary study on the effects of exercising to maximum walking distance on platelet and endothelial function in patients with intermittent claudication. AB - BACKGROUND: Platelet and endothelial activation has been shown to be increased in patients with intermittent claudication (IC). Recent studies have suggested that exercise may induce further platelet activation. The aims of this study were to investigate the effect of exercising to maximum walking distance on platelet and endothelial function in patients with intermittent claudication who were receiving statin and aspirin therapy compared with age matched healthy controls. METHODS: Platelet aggregation through COX-mediated and thrombin receptor activator peptide (TRAP)-stimulated GPIIb/IIIa pathways was measured by the Ultegra point of care system in 20 patients with IC on aspirin and 20 healthy volunteers before, immediately and 1h after exercising to treadmill maximal walking distance (MWD). Soluble P-selectin, vWF and sICAM were measured using an enzyme linked immuno-sorbent assay technique. RESULTS: Baseline platelet aggregation was significantly reduced in patients with IC compared to volunteers (p<0.05). In patients, exercising to MWD significantly reduced platelet aggregation (COX, median -5% [range -24 to 13%]; p = 0.02; GPIIIa/IIb, median 13% [range -72 to 33%]; p = 0.02) immediately post-exercise which returned to baseline values at 1 h. There was no change in the healthy volunteers following the same median duration of exercise. Baseline sP-selectin levels were higher in the patients with IC compared to the healthy volunteers [Median values (interquartile range), 42.72 (33.28-54.24) versus 29.16 (24.40-34.10), p = 0.0003] but there were no differences in vWF levels. Both sP-selectin and vWF levels increased significantly in the control and patient group following exercise (p<0.005). sICAM were higher at baseline in the patients with IC but were unchanged following exercise [Median values (interquartile range),560.9 (405.5-739.4) versus 467.0 (325.7-643.4), p<0.05]. CONCLUSION: This study is the first to show that platelet aggregation is reduced immediately following treadmill exercise to maximum walking distance in patients with IC despite a rise in sP-selectin and vWF, suggesting endothelial activation. The inhibition of platelet aggregation after exercise in subjects on antiplatelet and statin therapy suggests that exercise is unlikely to exacerbate platelet thrombus formation in patients with IC. PMID- 16360328 TI - Efficacy of vancomycin, teicoplanin and fusidic acid as prophylactic agents in prevention of vascular graft infection: an experimental study in rat. AB - OBJECTIVES: To compare the efficacy of a single prophylactic dose of intra peritoneal vancomycin and teicoplanin with anti-biotic treated Dacron grafts (vancomycin, teicoplanin, 10 or 40% fusidic acid-soaked grafts) in preventing vascular graft infections in a rat model. DESIGN: Prospective, randomized, controlled animal study. MATERIALS AND METHODS: The graft infections were established in the subcutaneous tissues of 80 female Sprague-Dawley rats by the implantation of Dacron prostheses followed by the topical inoculation with methicillin-resistant Staphylococcus aureus. The study groups were as follows: (1) uncontaminated control group, (2) untreated contaminated group, (3) contaminated group with intra-peritoneal vancomycin, (4) contaminated group with intra-peritoneal teicoplanin, (5) contaminated group received vancomycin-soaked Dacron graft, (6) contaminated group received teicoplanin-soaked Dacron graft, (7) contaminated group received 40% fusidic acid-soaked Dacron graft, and (8) contaminated group received 10% fusidic acid-soaked Dacron graft prophylaxis. The grafts were removed after 7 days and evaluated by a quantitative culture analysis. RESULTS: No infection was detected in controls. The untreated contaminated group had a high bacteria count (6.0 x 10(4) CFU/cm2 Dacron graft). Groups that received intra-peritoneal vancomycin or teicoplanin had less bacterial growth (4.8 x 10(3) and 3.9 x 10(3)CFU/cm2 Dacron graft, respectively). Similarly, the group that received 10% fusidic acid-soaked graft showed less bacterial growth (3.6 x 10(3) CFU/cm2 Dacron graft). The groups with vancomycin-, teicoplanin- and 40% fusidic acid-soaked grafts showed no evidence of infection. Statistical analyses demonstrated that intra-peritoneal prophylactic antibiotic treatment was less effective in inhibiting bacterial growth than high concentration antimicrobial-soaking of grafts. CONCLUSION: The use of vancomycin , teicoplanin- and 40% fusidic acid-soaked grafts was effective in preventing primary prosthetic vascular graft infection. PMID- 16360329 TI - In vitro dual effect of arsenic trioxide on hemopoiesis: inhibition of erythropoiesis and stimulation of megakaryocytic maturation. AB - Although the arsenic compounds are now widely utilized in clinics in the treatment of various tumors, their effects on normal hematopoiesis do not seem to have been explored. In the present study, we provide evidence that arsenic trioxide (As(2)O(3)) exerts in vitro a potent inhibitory effect on normal erythropoiesis and a stimulatory action on megakaryocytic differentiation. The effect of As(2)O(3) on erythroid and megakaryocytic differentiation was evaluated on both erythroleukemic cell lines K562 and HEL and on normal hemopoietic progenitor cells (HPCs) induced to selective erythroid or megakaryocytic differentiation. The inhibitory effect of As(2)O(3) on erythropoiesis is related to: (a) the inhibition of Stat5 activation with consequent reduced expression of the target genes Bcl-X(L) and glycophorin-A; (b) the activation of an apoptotic mechanism that leads to the cleavage of the erythroid transcription factors Tal-1 and GATA-1, whose integrity is required for erythroid cell survival and differentiation; (c) the reduced expression of heat shock protein 70, required for GATA-1 integrity. The stimulatory effect of As(2)O(3) on normal megakaryocytopoiesis is seemingly related to upmodulation of GATA-2 expression and to stimulation of MAPK activity. These observations may have implications for the patients undergoing anti-leukemic treatment with this compound. PMID- 16360330 TI - Distance measurements in disodium ATP hydrates by means of 31P double quantum two dimensional solid-state NMR spectroscopy. AB - POST-C7 measurements provide constraints allowing distinguishing crystal lattice organization and establishing intra and/or intermolecular distances between phosphorus atoms of triphosphate chains for different hydrates of disodium ATP salts. Double-quantum efficiency in function of excitation time obtained from series of two-dimensional spectra for POST-C7 experiments was used to set up of buildup curves and semi-quantitative measure of 31P-31P length. PMID- 16360331 TI - Pharmacologic characteristics of prulifloxacin. AB - Prulifloxacin, the prodrug of ulifloxacin, is a broad-spectrum oral fluoroquinolone antibacterial agent. After absorption, prulifloxacin is metabolized by esterases to ulifloxacin. The drug has a long elimination half life, allowing once-daily administration. In vitro, ulifloxacin is generally more active than other fluoroquinolones against a variety of clinical isolates of Gram negative bacteria, including community and nosocomial isolates of Escherichia coli, Klebsiella spp., Proteus, Providencia and Morganella spp., Moraxella catarrhalis, Haemophilus spp., and Pseudomonas aeruginosa. Gram-positive organisms, including methicillin- or oxacillin-susceptible Staphylococcus aureus, Enterococcus spp. and Streptococcus pneumoniae, are susceptible to ulifloxacin. In well-designed clinical trials, prulifloxacin showed good clinical and bacteriological efficacy (similar to that of ciprofloxacin) and was generally well tolerated, demonstrating a similar tolerability profile to that of ciprofloxacin. In conclusion, the in vitro inhibitory and bactericidal activities exhibited by ulifloxacin and the favorable characteristics shown by its prodrug (prulifloxacin) in clinical trials, particularly indicate this drug for the treatment of lung and urinary infections. PMID- 16360332 TI - A complication of brucellosis: epididymoorchitis. AB - BACKGROUND: Epididymoorchitis is the most frequent genitourinary complication of brucellosis. METHODS: This prospective study was conducted between February 2001 and January 2004, prospectively. Male patients diagnosed with brucellosis were included in this study and evaluated for testicular involvement. RESULTS: Epididymoorchitis was detected in 17 out of 134 (12.7%) male patients with brucellosis. Mean age of the patients was 36.9+/-7.1 years. Twelve patients (70.6%) had acute, four patients (23.5%) had subacute, and one patient (5.9%) had chronic brucellosis. The most common symptoms were scrotal pain (94%) and swelling (82%). Eleven patients had unilateral epididymoorchitis, four had unilateral orchitis and two had unilateral epididymitis. A testicular abscess was detected in one patient. Sperm analysis was performed on 14 patients. Five patients had aspermia and eight had oligospermia. Combined antibiotic therapy was started and continued for 6-8 weeks. Orchiectomy was required for two patients and granulomatous orchitis was detected in the resected specimens. Relapse occurred in only one patient. Three patients had permanent oligospermia and one patient had permanent aspermia after the antibiotic therapy. Younger age, high C reactive protein level and blood culture positivity were statistically significant differences between the patients with and without epididymoorchitis. CONCLUSIONS: Brucellosis should be considered in the diagnosis of scrotal diseases in endemic areas. A conservative approach is usually adequate for managing brucellar epididymoorchitis. However, infertility problems may develop in these patients. Well-designed further investigations are needed to explain the relationship between brucellar epididymoorchitis and infertility in man. PMID- 16360333 TI - Ultrasonic frequency effects on the removal of Microcystis aeruginosa. AB - Algae bloom in source water causes high chemical consumption and deteriorates water quality in waterworks. This paper studied the ultrasonic removal of Microcystis aeruginosa. The results showed that algae cells could be effectively removed by sonication and gas vesicle collapse was the main mechanism. The ultrasonic algae removal followed the first order reaction with a rate constant of 0.023 min(-1) (80 W, 80 kHz). Higher ultrasound frequency benefited algae removal; the algae removal rate constant was 0.114 min(-1) at 1320 kHz and 0.0224 min(-1) at 20 kHz (80 W). Higher ultrasound power also accelerated algae removal; the algae removal rate constant was 0.023 min(-1) at 80 W and 0.007 min(-1) at 32 W (80 kHz). However, high ultrasound power and long irradiation caused microcystins to increase. 80 W, 80 kHz sonication for 5 min increased the extracellular microcystins concentration from 0.87 microg/L to 3.11 microg/L. Sound frequency had little impact on the microcystins release. The chlorophyll a concentration initially decreased and then stabilized after 5 min of sonication. PMID- 16360334 TI - Apoptosis of macrophages induced by Trichomonas vaginalis through the phosphorylation of p38 mitogen-activated protein kinase that locates at downstream of mitochondria-dependent caspase activation. AB - Trichomonas vaginalis, a flagellated protozoan parasite, is the causative organism of trichomoniasis. We have recently demonstrated that T. vaginalis induces apoptotic cell death via a Bcl-x(L)-dependent pathway in RAW264.7 macrophages. In this study, we attempted to characterize in detail the signaling cascades resulting in T. vaginalis-induced macrophage apoptosis, focusing particularly on mitochondrial changes and the role of p38 mitogen-activated protein kinase (p38 MAPK) activation. We found that T. vaginalis induced mitochondrial changes including the release of cytochrome c and the serial activation of caspases, leading to the activation of p38 MAPK in macrophages. These biochemical changes culminated in the apoptosis of the host cells. Caspase inhibitors induced a significant inhibition of T. vaginalis-induced nuclear damage, as well as the activation of p38 MAPK. Treatment with the p38 MAPK inhibitor, SB203580, or the overexpression of kinase-inactive p38 MAPK, induced an attenuation of T. vaginalis-induced apoptosis but not cytochrome c release, the activation of caspase-9 and caspase-3, or PARP cleavage. Furthermore, SB203580 treatment to human macrophages consistently blocked T. vaginalis-induced apoptosis. Collectively, our findings indicate that p38 MAPK signaling cascade is requisite to apoptosis of T. vaginalis-infected macrophage, and this apoptotic process occurs via the phosphorylation of p38 MAPK, which is located downstream of mitochondria-dependent caspase activation, conferring insight into the plausible molecular mechanism of T. vaginalis-immune evasion from macrophage attack. PMID- 16360335 TI - Survival strategy of Echinococcus multilocularis in the human host. AB - As exemplified by "aborted" calcified liver lesions commonly found in patients from endemic areas, Echinococcus multilocularis metacestodes develop only in a minority of individuals exposed to infection with the papasite. Clinical research has disclosed some aspects of the survival strategy of E. multilocularis in human hosts. Clinical observations in liver transplantation and AIDS suggest that suppression of cellular/Th1-related immunity increases disease severity. Most of the studies have stressed a role for CD8+ T cells and for Interleukin-10 in the development of tolerance. A spontaneous secretion of IL-10 by the PBMC seems to be the immunological hallmark of patients with progressive forms of alveolar echinococcosis (AE). IL-10-induced inhibition of effector macrophages, but also of antigen-presenting dendritic cells, may be operating and allowing parasite growth and survival. The genetic correlates of susceptibility to infection with E. multilocularis are clearer in humans than in the mouse model. A significant link between MHC polymorphism and clinical presentation of AE has been shown, and the spontaneous secretion of IL-10 in patients with a progressive AE is higher in patients with the HLA DR3+, DQ2+ haplotype. Clustering of cases in certain families, in communities otherwise exposed to similar risk factors, also points to immuno-genetic predisposition factors that may allow the larva to escape host immunity more easily. The first stage of larval development may be crucial in producing "danger signals" stimulating the initial production of cytokines. Therapeutic use of Interferon alpha is an attempt to foil the survival strategy of E. multilocularis. PMID- 16360336 TI - Recent advances in characterization of Echinococcus antigen B. AB - Antigen B (AgB) in hydatid cyst fluid of Echinococcus granulosus is a polymeric lipoprotein of 160 kDa and a highly immunogenic major antigen in echinococcal infection. The antigen is comprised of a group of subunit monomers of approximately 8 kDa in molecular size. Recent studies have revealed that the E. granulosus AgB (EgAgB) shows a high degree of genetic variability and the genes encoding the EgAgB 8-kDa subunit monomers that have been identified to date could be grouped into four clades, corresponding to the genes EgAgB8/1, EgAgB8/2, EgAgB8/3 and EgAgB8/4. It has been suggested that the recombinant EgAgB8/2 (rEgAgB8/2) provides better performance in serodiagnosis of human cystic echinococcosis (CE) than does the recombinant EgAgB8/1 (rEgAgB8/1). The EgAgB has been identified as a protease inhibitor with an ability to inhibit recruitment of neutrophils and exploit activation of T helper cells by eliciting a non protective Th2 cell response, predominantly in patients with progressive CE. Recently it has been revealed that AgB also exists in the cyst fluid of Echinococcus multilocularis. Five different cDNAs encoding the EgAgB homologues have been identified in vesicles, protoscoleces and/or immature adult worms of E. multilocularis and named as EmAgB8/1, EmAgB8/2, EmAgB8/3, EmAgB8/4 and EmAgB8/5. These genes appeared to be expressed in a developmentally regulated manner in the parasite life cycle. This review focuses on recent advances in molecular biological and immunological characterization of AgB from both of E. granulosus and E. multilocularis. PMID- 16360337 TI - Mitochondrial DNA diagnosis for taeniasis and cysticercosis. AB - Molecular diagnosis for taeniasis and cysticercosis in humans on the basis of mitochondrial DNA analysis was reviewed. Development and application of three different methods, including restriction fragment length polymorphism analysis, base excision sequence scanning thymine-base analysis and multiplex PCR, were described. Moreover, molecular diagnosis of cysticerci found in specimens submitted for histopathology and the molecular detection of taeniasis using copro DNA were discussed. PMID- 16360338 TI - Spontaneous elimination of hepatitis C virus RNA in individuals with persistent infection in a hyperendemic area of Japan. AB - The natural course of hepatitis C virus (HCV) carriers is not well understood. We examined the clinical characteristics of individuals exhibiting spontaneous elimination of HCV as part of a cohort study of residents of a HCV hyperendemic area in Japan. In individuals who were judged to have persistent HCV infection in 1995, 302 had at least 4 annual ALT measurements between 1993 and 2000, and had not been treated with IFN. They were tested for the presence of HCV RNA in 2001 and/or 2002 and HCV RNA could not be detected in 20 of the 302 individuals. In these 20 individuals, 7 were confirmed to have detectable HCV RNA and 13 were not until 2000. Thus, 2.4% (7/289) were judged to have spontaneously eliminated the HCV infection during that 6-year period. Although there were no differences in age, sex, ALT levels, or serologically defined HCV genotype between individuals with and without exhibiting spontaneous elimination, there was a significant relationship between the elimination of HCV RNA and a low level of HCVcAg (<20pg/mL) (P<0.001) upon testing in 1995. These results suggest that spontaneous elimination of HCV RNA following persistent infection is rare and appears to be related to viral load. PMID- 16360339 TI - The genetic portrait of an outbreak strain. AB - Unique events in the genome of Mycobacterium tuberculosis, including deletions and IS6110 insertions, have been proposed to be responsible for the virulence phenotype of outbreak strains. Based on this premise, we determined ten IS6110 insertion sites in the genome of the M. tuberculosis CH strain, which was responsible for a large outbreak in Leicestershire, England. Together with previous data on genomic deletions, it was found that 16 genes were mutated either by IS6110 insertions or deletions. The likely impact of these genomic events on the phenotype of the CH strain is discussed. PMID- 16360340 TI - Characterization of the immunological features of tuberculous pericardial effusions in HIV positive and HIV negative patients in contrast with non tuberculous effusions. AB - OBJECTIVE: To investigate the immunopathogenesis of pericardial tuberculosis (TB) and the influence of human immunodeficiency virus (HIV) on the anti-tuberculous immune response. DESIGN: Consecutive patients presenting with large pericardial effusions were subjected to a full clinical examination and pericardiocentesis. Aspirated fluid was sent for biochemistry, differential leukocyte count, flow cytometric analysis and determination of cytokine levels. Pericardial tissue was sent for TB culture and histopathological evaluation. Diagnoses were made according to pre-determined criteria. RESULTS: Fifty-six patients were included and divided into HIV positive TB (n = 22), HIV negative TB (n = 21) and non tuberculous effusions (n = 13). Peripheral blood neutrophil, lymphocyte and monocyte counts were significantly lower in HIV positive TB patients. Lymphocytes were the dominant cell type in tuberculous pericardial effusions. CD4+ cells dominated in HIV negative tuberculous effusions, whereas CD8+ cells dominated in HIV positive TB. The difference in the concentration of IFN-gamma levels in the tuberculous and non-tuberculous pericardial effusions was statistically significant. Despite significant differences in pericardial CD4+ cell counts, IFN gamma levels were similarly elevated in HIV negative and HIV positive tuberculous effusions. Highest levels of pericardial IL-10 were observed in samples associated with least tissue necrosis, suggesting the possibility of a tissue protective immunoregulatory role for IL-10. CONCLUSIONS: Tuberculous pericardial effusions result from a T helper1 (Th1)-dominant immune response. IFN-gamma producing CD4+ lymphocytes dominate in HIV negative patients, whereas CD8+ seem to play a more important role in HIV positive patients. Infection with HIV leads to the depletion of immunocompetent cells such as monocytes, NK cells and neutrophils. PMID- 16360341 TI - A partial down-regulation of WASP is sufficient to inhibit podosome formation in dendritic cells. AB - The Wiskott Aldrich syndrome protein (WASP) is a hematopoietic-specific cytoskeletal regulator that is necessary for induction of normal immunity. In the context of effective gene therapy for WAS, cellular models of human WASP deficiency are important for definition of the threshold of protein expression required for optimal activity. Using lentiviral vector-mediated RNA interference (RNAi), we were able to down-regulate the levels of human WASP in cell lines and primary cells. In dendritic cells (DC), RNAi-induced WASP deficiency did not impair phenotypic maturation but perturbed cytoskeletal organization. As a result, podosomes, which are actin-rich structures present in immature adherent DC, were formed less efficiently and motility was disturbed. Overall, treatment of cells with RNAi recapitulated the phenotype of cells derived from patients or animals with inactivating mutations of the WAS gene. Interestingly, reduction of the levels of WASP to about 60% of normal was sufficient to inhibit the formation of podosomes in DC, implying that this cell type requires near-normal levels of WASP to sustain physiological cytoskeleton-dependent activities. PMID- 16360342 TI - Terminology model discovery using natural language processing and visualization techniques. AB - Medical terminologies are important for unambiguous encoding and exchange of clinical information. The traditional manual method of developing terminology models is time-consuming and limited in the number of phrases that a human developer can examine. In this paper, we present an automated method for developing medical terminology models based on natural language processing (NLP) and information visualization techniques. Surgical pathology reports were selected as the testing corpus for developing a pathology procedure terminology model. The use of a general NLP processor for the medical domain, MedLEE, provides an automated method for acquiring semantic structures from a free text corpus and sheds light on a new high-throughput method of medical terminology model development. The use of an information visualization technique supports the summarization and visualization of the large quantity of semantic structures generated from medical documents. We believe that a general method based on NLP and information visualization will facilitate the modeling of medical terminologies. PMID- 16360343 TI - Energy cost of activity and exercise in children and adolescents with cystic fibrosis. AB - In cystic fibrosis (CF), perturbations of total daily energy expenditure (TDEE) may be a major determinant of altered nutrition and growth. Measurement of TDEE is problematic, though the flex-heart rate method (FHRM) provides a close estimation of TDEE, as compared to the cost-prohibitive, gold standard, the double-labeled water method, and permits estimates of the energy cost of daily activities (ECA) above resting energy expenditure (REE). We hypothesize that alterations in ECA affects TDEE in CF. PURPOSE: To measure components of TDEE in adolescents with CF and normal lung function compared with controls, and to determine whether ECA can be improved by diet and exercise. METHODS: Clinically stable CF subjects (aged 9-13, n=12) and age- and gender-matched controls (n=13) had repeated measurements of TDEE by FHRM, REE, and maximal cardiopulmonary exercise testing (CPET) during a 6-week exercise and diet program. RESULTS: While the mean REE was similar in both groups, ECA was significantly lower in CF adolescents as compared to controls (p=0.02). During CPET, maximal exercise in CF was characterized by hyperventilation, which was unrelated to ventilation perfusion mismatching. There were no changes in REE after dietary intervention. CONCLUSION: ECA in CF adolescents with normal lung function is lower when compared to healthy controls. These findings support the hypothesis that clinically stable patients with CF have inefficient energy metabolism or alternatively conserve energy during activities of daily living. PMID- 16360344 TI - Liquid chromatography-electrospray mass spectrometry determination of free and total concentrations of ropivacaine in human plasma. AB - A specific and sensitive liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method was developed for the determination of free and total ropivacaine in human plasma. The work-up procedure involved a simple precipitation of plasma proteins with methanol. Etidocaine served as the internal standard. After microscale equilibrium-dialysis, measurement of free ropivacaine levels was performed after direct injection of the dialysate into the chromatograph. The system used a Zorbax eclipse XD8 C8 analytical column packed with 5 microm diameter particles as the stationary phase. The mobile phase consisted of a 15-min gradient (mobile phase A: 0.05% (v/v) trimethylamine in acetonitrile, mobile phase B: 2mM ammonium formate buffer (pH 3)). Mass spectrometric data were acquired in single ion monitoring mode at m/z 275 for ropivacaine and m/z 277 for etidocaine. The drug/internal standard peak area ratios (plasma) or peak areas (dialysate) were linked via a quadratic relationship to concentrations. Precision ranged from 1 to 7.6% accuracy was between 92.6 and 109%. The lower limits of quantitation were 1 microg/l in plasma and 2 microg/l in the dialysate. This method was found suitable for the analysis of plasma samples collected during a clinical trial performed in 30 infants undergoing epidural anaesthesia or continuous psoas compartment block. PMID- 16360345 TI - Effect of vasopressin on hemodynamics in patients with refractory cardiogenic shock complicating acute myocardial infarction. AB - In a retrospective study of 36 patients who developed cardiogenic shock after myocardial infarction, intravenous vasopressin therapy increased mean arterial pressure from 56 to 73 mm Hg at 1 hour (p < 0.001) and maintained it for 24 hours without changing pulmonary capillary wedge pressure, cardiac index, urine output, or other inotropic requirements. After norepinephrine administration, mean pulmonary capillary wedge pressure increased at 1 hour from 21 to 24 mm Hg (p = 0.04); however, this increase was not sustained at 12 and 24 hours. Norepinephrine was associated with a significant increase in cardiac power index at 24 hours, whereas there was only a trend for an increase in cardiac power with vasopressin therapy. In a cohort of patients who developed refractory cardiogenic shock after myocardial infarction, vasopressin was associated with increased mean arterial pressure and no adverse effect on other hemodynamic parameters. PMID- 16360346 TI - Relation of myocardial perfusion defects and nonsignificant coronary lesions by angiography with insights from intravascular ultrasound and coronary pressure measurements. AB - Several studies have demonstrated a correlation between myocardial ischemia and severity of coronary lesions as determined by intravascular ultrasound (IVUS) and fractional flow reserve (FFR) measurements. However, their value for the assessment of mild coronary stenoses that are associated with myocardial perfusion abnormalities has not been well studied. The objective of this study was to prospectively compare the results of myocardial perfusion as determined by exercise/dipyridamole myocardial single-photon emission computed tomography with IVUS and FFR measurements in patients who had angiographically mild coronary stenosis (< 50% diameter stenosis by quantitative coronary angiography). Forty eight patients who had stable coronary disease (61 +/- 11 years of age; 6 women) were included. All had mild coronary stenosis in the proximal/middle segment of > or = 1 coronary artery and had undergone maximal exercise myocardial technetium 99m tetrofosmin single-photon emission computed tomography within 48 hours before coronary angiography. IVUS measurements included lesion lumen area, external elastic membrane area, lesion plaque burden (calculated as external elastic membrane minus lumen area, divided by external elastic membrane, and multiplied by 100), and lumen area stenosis (calculated as reference lumen area minus lesion lumen area, divided by reference lumen area, multiplied by 100). Fifty-three coronary lesions were studied, with a mean percent diameter stenosis of 34.9 +/- 7.9% on angiography. Myocardial perfusion defects were demonstrated by single photon emission computed tomography in 11 patients (12 myocardial regions) with no differences in lesion percent diameter stenosis compared with those without perfusion defects. The presence of reversible perfusion defects was associated with a higher lesion plaque burden as evaluated by IVUS (67.4 +/- 8.1% vs 60.2 +/ 9.3%, p = 0.01). FFR values did not differ in the presence or absence of perfusion defects (0.90 +/- 0.06 vs 0.92 +/- 0.07, respectively; p = NS). In conclusion, plaque burden as determined by IVUS may partly explain the presence of myocardial perfusion defects in cases of angiographically nonsignificant coronary lesions. However, the high FFR values associated with these lesions suggest that other mechanisms, such as endothelial/microvascular dysfunction, might also account for perfusion abnormalities in these patients. PMID- 16360347 TI - Effect of phentolamine on the hyperemic response to adenosine in patients with microvascular disease. AB - For accurate measurement of the fractional flow reserve (FFR) of the myocardium, the presence of maximum hyperemia is of paramount importance. It has been suggested that the hyperemic effect of the conventionally used hyperemic stimulus, adenosine, could be submaximal in patients who have microvascular dysfunction and that adding alpha-blocking agents could augment the hyperemic response in these patients. We studied the effect of the nonselective alpha blocking agent phentolamine, which was administered in addition to adenosine after achieving hyperemia, in patients who had microvascular disease and those who did not. Thirty patients who were referred for percutaneous coronary intervention were selected. Of these 30 patients, 15 had strong indications for microvascular disease and 15 did not. FFR was measured using intracoronary adenosine, intravenous adenosine, and intracoronary papaverine before and after intracoronary administration of the nonselective alpha blocker phentolamine. In patients who did not have microvascular disease, no differences in hyperemic response to adenosine were noted, whether or not alpha blockade was given before adenosine administration; FFR levels before and after phentolamine were 0.76 and 0.75, respectively, using intracoronary adenosine (p = 0.10) and 0.75 and 0.74, respectively, using intravenous adenosine (p = 0.20). In contrast, in patients who had microvascular disease, some increase in hyperemic response was observed after administration of phentolamine; FFR levels decreased from 0.74 to 0.70 using intracoronary adenosine (p = 0.003) and from 0.75 to 0.72 using intravenous adenosine (p = 0.04). Although statistically significant, the observed further decrease in microvascular resistance after addition of phentolamine was small and did not affect clinical decision making in any patient. In conclusion, when measuring FFR, routinely adding an alpha-blocking agent to adenosine does not affect clinical decision making. PMID- 16360348 TI - Cardiovascular events in patients with coronary plaque rupture and nonsignificant stenosis. AB - The long-term outcome of patients who present with an identified plaque rupture in a nonflow-limiting lesion is not well known. We retrospectively studied 17 consecutive patients in whom intravascular ultrasound identified plaque rupture in nonflow-limiting lesions (minimum lumen area >4.0 mm2) that were not treated with coronary intervention. After a mean follow-up of 43 +/- 25 months, events related to those lesions were 1 death (6%) of undetermined cause (6%) that occurred after 69 months, no myocardial infarction, and 2 revascularizations (12%) that occurred at 3 and 67 months. Overall, the cumulative rate of cardiac events was 18%. PMID- 16360349 TI - Effect of atorvastatin (80 mg/day) versus pravastatin (40 mg/day) on arterial remodeling at coronary branch points (from the REVERSAL study). AB - The effect of moderate and intensive lipid lowering on plaque progression and arterial remodeling at coronary branch points was investigated. Intensive (+1 +/- 19.6%), but not moderate (+4.1 +/- 15.1%), lipid lowering prevented an increase in the percent [corrected] plaque area at the branch points. The 2 strategies were associated with increased areas of the lumen (+7.6% to 9.4%) and external elastic membrane (+9.6% to 10.8%). In contrast, there was no significant change in plaque, lumen, and/or external elastic membrane areas at the nonbranch point site. These results suggest that intensive lipid lowering can have a dramatic effect on atheroma-prone regions and that remodeling in response to changes in plaque is a heterogenous process. PMID- 16360350 TI - Comparison of serum lipid values in patients with coronary artery disease at <50, 50 to 59, 60 to 69, and >70 years of age. AB - We evaluated the relation between lipids and precocity of coronary artery disease (CAD) in the real world as characterized by increasing statin use. The highest mean values of total cholesterol, low-density lipoprotein cholesterol, triglycerides, non-high-density lipoprotein (HDL) cholesterol, and ratio of triglycerides to HDL cholesterol were found when CAD was detected in patients who were <50 years of age (p <0.01 for all); the opposite occurred for HDL cholesterol (p <0.01). Triglycerides and ratio of triglycerides to HDL cholesterol were the most powerful, independent variables related to precocity of CAD. PMID- 16360351 TI - Clinical characteristics and prognostic factors in patients with complicated acute coronary syndromes requiring prolonged mechanical ventilation. AB - Patients with acute coronary syndromes (ACSs) may develop serious multiorgan complications and require prolonged intensive care. Our aim was to characterize and identify factors that are associated with outcomes in these patients. We retrospectively identified 267 consecutive patients admitted to the coronary care unit for an ACS who required >3 days of mechanical ventilation. Multiple clinical and laboratory variables were correlated with mortality. Patients' ages were 68.3 +/- 10.9 years (mean +/- SD) and 165 (62%) were men. Seventy-six patients (29%) died within 30 days of admission, and the 1 year mortality was 46%. Moderate or severe left ventricular systolic dysfunction was found in 72% of the patients. Eighty-nine patients (33.3%) required vasopressors, of whom 64 (72%) did not survive 30 days. Among 127 patients who required antibiotics (48.3%), 30-day mortality was 53% compared with 4% among patients who did not require antibiotics (p <0.001). The 30-day mortality among patients who received both antibiotics and vasopressors was 64 of 87 patients (74%), and the 1-year mortality in this subgroup was 86.2%. Parameters found to be independent predictors of 30-day mortality were (in descending order): vasopressor requirement, use of antibiotics, peripheral vascular disease, ST-elevation myocardial infarction, renal failure, obesity and Killip class on admission. In conclusion, mortality among patients who require prolonged mechanical ventilation after an ACS is substantial. The main independent predictors of with mortality are the severity of heart failure and the presence of co-morbidities. PMID- 16360352 TI - Outcomes of patients consented but not randomized in a trial of primary percutaneous coronary intervention in acute myocardial infarction (the CADILLAC registry). AB - Baseline features, management, and outcomes of patients who had acute myocardial infarction (AMI) and were excluded from randomized trials of primary percutaneous coronary intervention (PCI) have not been well described. We examined the baseline features and outcomes of patients who had AMI and were excluded due to angiographic ineligibility from a randomized trial of primary PCI. The CADILLAC trial evaluated 4 primary PCI strategies in patients who had AMI without cardiogenic shock. Of 2,681 patients who consented, 599 (22.3%) were subsequently excluded from randomization due to protocol-specified angiographic findings. These patients were enrolled in a formal in-hospital registry and were treated at the discretion of attending physicians. Registry versus randomized patients were older (median age 61.9 vs 59.0 years, p = 0.002), more frequently had 3-vessel disease (52.4% vs 15.6%, p = 0.0001), and more frequently had lower left ventricular ejection fraction (45% vs 50%, p = 0.002). Registry patients were treated with PCI (n = 234), coronary artery bypass grafting (n = 136), or medically only (n = 226). In-hospital mortality rate among all patients who consented was 2.2% and was higher among registry than among randomized patients (4.0% vs 1.6%, relative risk 2.45, p = 0.001). Within the registry, covariate adjusted risk of mortality was lower among patients who were treated by PCI compared with those who were treated medically (odds ratio 0.21, p=0.03). In conclusion, angiographic ineligibility for randomization in a large, prospective, primary PCI trial was associated with numerous high-risk demographic characteristics and higher short-term mortality. Nonetheless, the overall survival of patients who were referred for primary PCI was excellent, and performance of primary PCI versus medical therapy was associated with increased survival among excluded patients. PMID- 16360353 TI - Comparison between sirolimus-eluting stents and intracoronary catheter-based beta radiation for the treatment of in-stent restenosis. AB - We report the outcomes of patients who had in-stent restenosis (IRS) that was treated with intravascular brachytherapy (IVBT) or sirolimus-eluting stent (SES) implantation. The benefit of IVBT for treating ISR is well documented. SES implantation decreases first-time ISR and, in preliminary reports, has been used to treat ISR. Fifty consecutive patients who had ISR were treated; the first 25 patients underwent SES implantation and the next 25 patients were treated with IVBT using a beta-Cath System (a 40-mm strontium-90/yttrium-90 source). Quantitative angiographic and intravascular ultrasound follow-up were performed at 5.2 +/- 1.1 and 12.1 +/- 1.2 months; clinical follow-up was performed at 15 months. SES deployment and IVBT were successful in all patients. At 12-month follow-up, 8 patients who underwent IVBT had angiographic recurrence (4 in the stent and 4 at the stent edge); only 1 patient who underwent SES implantation developed recurrent ISR. At 12 months, in-stent late luminal loss was similar between the SES and IVBT groups (0.35 +/- 0.45 vs 0.34 +/- 0.46 mm, p = 0.9); however, in-stent net luminal gain was higher in the SES group than in the IVBT group (1.32 +/- 0.13 vs 0.57 +/- 0.19 mm, p <0.0001), and in-lesion late luminal loss was higher in the IVBT group (0.48 +/- 0.32 vs 0.16 +/- 0.42 mm, p = 0.004). At 12 months, intravascular ultrasound stent volume obstruction was higher after IVBT versus than after SES implantation (38.7% vs 6.7%, p <0.0001). At 15-month clinical follow-up, 64% and 96% (p <0.01) of patients who underwent IVBT and SES implantation, respectively, were free of major adverse cardiac events. In conclusion SES implantation for the treatment of ISR was effective and superior to catheter-based IVBT in preventing recurrent neointimal proliferation and angiographic restenosis at 1-year follow-up. PMID- 16360355 TI - Usefulness of 4-, 8-, and 16-slice computed tomography for detection of graft occlusion or patency after coronary artery bypass grafting. AB - The sensitivity and specificity of computed tomographic angiography for the evaluation of coronary artery bypass grafts (CABGs) was reviewed. A search of published studies in all languages was performed, incorporating electronic and manual components. A total of 985 patients with 2,200 CABGs participated in investigations with single-detector computed tomography (CT), 441 patients (1,246 CABGs) with 4-slice CT, none with 8-slice CT, and 144 patients (416 grafts) with 16-slice CT. Pooled data showed a sensitivity for the detection of complete occlusion with single-slice CT of 81% (402 of 494 patients), with 4-slice CT of 93% (293 of 315 patients), and with 16-slice CT of 99% (75 of 76 patients). The specificity, based on pooled data, using single-slice CT was 89% (1,507 of 1,697 patients), with 4-slice CT was 96% (878 of 915 patients), and with 16-slice CT was 98% (301 of 306 patients). The detection of complete occlusion and the detection of patency in saphenous vein bypass grafts and arterial bypass grafts were similar with multislice CT. Limited data were available on the detection of significant stenosis, exclusive of complete occlusion. With 4-slice CT, the sensitivity was 74% (23 of 31 patients) and was 88% (21 of 24 patients) with 16 slice CT. In conclusion, single-detector CT was able to detect graft patency, but it was not sensitive for graft occlusion. The data suggest that 4- and 16-slice CT can be used for the detection of complete graft occlusion or graft patency of CABGs. Significant stenosis was better assessed with 16-slice CT than with 4 slice CT. PMID- 16360354 TI - Clinical outcome following aleatory implantation of paclitaxel-eluting or sirolimus-eluting stents in complex coronary lesions. AB - We compared the clinical efficacy of paclitaxel-eluting stents (PESs) and sirolimus-eluting stents (SESs) in a contemporary cohort of patients who had complex lesions. We collected data on 9-month outcomes in 529 patients (281 in the PES group and 248 in the SES group) whose de novo lesions were treated with drug-eluting stents. The end point was per-patient in-hospital and follow-up major adverse cardiac events, which were defined as a composite of death, myocardial infarction, and target vessel revascularization, including target lesion revascularization. There were no in-hospital deaths or repeat revascularizations; however, 5.7% of the PES group and 2% of the SES group developed a myocardial infarction (p = 0.04). At a median follow-up of 10.6 months, the rate of major adverse cardiac events was similar between groups (18.1% vs 21%, adjusted hazard ratio 0.85, 95% confidence interval 0.57 to 1.25), without any difference in the occurrence of death or myocardial infarction. Diabetes and total stent length were independent predictors of major adverse cardiac events. Propensity analysis confirmed the similarity between devices (hazard ratio 0.87, 95% confidence interval 0.62 to 1.25). Most restenoses were focal and only 2 patients required surgical revascularization. In conclusion, implantation of drug-eluting stents in complex lesions was associated with favorable results and most patients remained free from surgical revascularization at follow-up. Overall, the 2 available stent platforms had similar performance characteristics. PMID- 16360356 TI - Implications from the Air Force/Texas Coronary Atherosclerosis Prevention Study for the Adult Treatment Panel III guidelines. AB - The Air Force/Texas Coronary Atherosclerosis Prevention Study (AFCAPS/TexCAPS) first reported its results in 1998, before the 2001 publication of the National Cholesterol Education Program-Adult Treatment Panel III guidelines (NCEP-ATP III) and 2004 update. Our objective was to investigate the impact of these guidelines on the AFCAPS/TexCAPS cohort. The main outcome measures were the event rates of first acute major coronary events (AMCEs), which were reduced 39% by lovastatin (95% confidence interval [CI] 21% to 53%, p <0.001) in the 65% of the cohort eligible for drug therapy and by 34% (95% CI -9% to 60%, p = 0.108) in the remaining 35% for whom drug therapy was considered optional. The evaluation of other guideline components included a 44% (95% CI 27% to 58%, p <0.001) reduction in AMCEs in subjects with baseline high-density lipoprotein cholesterol <40 mg/dl and a 41% (95% CI 19% to 57%) reduction in AMCEs in subjects with the metabolic syndrome. In the recent update, patients who had a moderately high risk of coronary heart disease and a baseline low-density lipoprotein cholesterol level of 100 to 130 mg/dl could be considered for therapy with a medication to lower the low-density lipoprotein cholesterol level to <100 mg/dl. A total of 334 subjects (5.1%) were in this group, in whom lovastatin reduced the risk of AMCEs by 68% (95% CI 12% to 88%, p = 0.027). However, 21% of the AMCEs were missed by the guidelines. Metabolic syndrome was noted in 48% of these subjects and may help define those in whom treatment with a medication is now considered optional. In conclusion, the ability of the ATP III guidelines and its update has markedly improved our ability to define coronary heart disease risk; however, other components of the guidelines, such as non-high-density lipoprotein cholesterol and the optional low-density lipoprotein cholesterol target goal of <100 mg/dl, still require additional evaluation. PMID- 16360357 TI - Premature coronary heart disease, cigarette smoking, and the metabolic syndrome. AB - The metabolic syndrome and cigarette smoking each increase the risk of a recurrent event in patients with premature coronary heart disease. We explored the association between cigarette smoking and the metabolic syndrome by examining 705 men aged < 55 years and 296 women < 65 years within 6 to 12 months of a major coronary heart disease event. Most were taking statins (96%) and antihypertensive drugs (88%). Nearly 1/3 of the subjects had the full metabolic syndrome, as defined by the National Cholesterol Education Program criteria. These subjects were less likely to be nonsmokers than were those with < or = 2 components of the metabolic syndrome (13.2% vs 24.2%, p < 0.0001). After adjustment for age, educational attendance, and alcohol consumption, the odds ratio (OR) for the metabolic syndrome was doubled in men who smoked cigarettes daily (OR 2.2, 95% confidence interval 1.3 to 3.7) or who were ex-smokers (OR 2.3, 95% confidence interval 1.4 to 3.9) compared with nonsmokers. Female ex-smokers had an increased risk compared with nonsmokers (OR 2.0, 95% confidence interval 1.0 to 3.9). Ex smokers were more likely to meet the metabolic syndrome cutoff levels for waist circumference and high-density lipoprotein cholesterol (p < or = 0.01) than were nonsmokers. Also, male ex-smokers were more likely to exceed the cutoff level for triglycerides (p = 0.004). These findings indicate that although smoking cessation is imperative for patients with premature CHD, the metabolic risks associated with overweight and obesity after cessation need to be addressed. PMID- 16360358 TI - Myocardial and vascular dysfunction and exercise capacity in the metabolic syndrome. AB - The metabolic syndrome (MS) is associated with cardiovascular risk exceeding that expected from atherosclerotic risk factors, but the mechanism of this association is unclear. We sought to determine the effects of the MS on myocardial and vascular function and cardiorespiratory fitness in 393 subjects with significant risk factors but no cardiovascular disease and negative stress echocardiographic findings. Myocardial function was assessed by global strain rate, strain, and regional systolic velocity (s(m)) and diastolic velocity (e(m)) using tissue Doppler imaging. Arterial compliance was assessed using the pulse pressure method, involving simultaneous radial applanation tonometry and echocardiographic measurement of stroke volume. Exercise capacity was measured by expired gas analysis. Significant and incremental variations in left ventricular systolic (s(m), global strain, and strain rate) and diastolic (e(m)) function were found according to the number of components of MS (p <0.001). MS contributed to reduced systolic and diastolic function even in those without left ventricular hypertrophy (p <0.01). A similar dose-response association was present between the number of components of the MS and exercise capacity (p <0.001) and arterial compliance. The global strain rate and e(m) were independent predictors of exercise capacity. In conclusion, subclinical left ventricular dysfunction corresponded to the degree of metabolic burden, and these myocardial changes were associated with reduced cardiorespiratory fitness. Subjects with MS who also have subclinical myocardial abnormalities and reduced cardiorespiratory fitness may have a higher risk of cardiovascular disease events and heart failure. PMID- 16360359 TI - Positron emission tomographic evaluation of regulation of myocardial perfusion in physiological (elite athletes) and pathological (systemic hypertension) left ventricular hypertrophy. AB - Myocardial perfusion (MP) may differ in physiologic and pathologic left ventricular hypertrophy (LVH). We compared MP in LVH in elite athletes and patients with hypertension with healthy, age-matched subjects. We included 12 rowers with LVH, 19 patients with hypertension with LVH, and 2 age-matched groups of healthy subjects (n = 11 and n = 12). The left ventricular mass index was determined echocardiographically. MP was measured by N-13 ammonia positron emission tomography. The maximal perfusion and perfusion reserve were studied using dipyridamole, and endothelial function was assessed by a cold pressor test. The degree of LVH was similar in athletes and those with hypertension. Compared with controls, athletes had 20% lower baseline MP (p <0.05), a similar response to the cold pressor test, and a higher perfusion reserve (31%, p <0.05). The patients with hypertension had a 25% higher baseline MP (p <0.05), a reduced increase during the cold pressor test (12% vs 25% in controls, p <0.05), and a reduced perfusion reserve (27% lower, p <0.001). The peak global perfusion (MP x left ventricular mass index) was 62% higher in athletes (p <0.05) than in controls, but the peak global perfusion in patients with hypertension did not differ from that of controls. In conclusion, physiologic LVH in athletes is suited for a high peak workload at the cost of only a small increase in basal myocardial oxygen consumption. In contrast, LVH in the presence of hypertension is a good adaptation to the increased baseline workload with maintained maximal cardiac performance. Endothelial dysfunction may contribute to the reduced perfusion reserve seen in hypertensive LVH. PMID- 16360361 TI - Relation of plasma brain natriuretic peptide levels in non-ST-elevation coronary disease and preserved systolic function to number of narrowed coronary arteries. AB - Elevated plasma brain natriuretic peptide (BNP) levels have been described in patients with acute myocardial infarction and left ventricular dysfunction. The aim of the present study was to evaluate circulating BNP levels in patients with coronary artery disease without ST-segment elevation acute myocardial infarction and preserved systolic function and to evaluate the BNP levels in relation to the number of involved coronary vessels. We studied 88 patients with coronary artery disease: group 1 had stable angina, group 2 had unstable angina (UA), group 3 had non-Q-wave myocardial infarction (NSTEMI), and group 0 consisted of 15 healthy subjects. All recruited subjects underwent angiographic examination and echocardiographic evaluation. No patients had heart failure, previous myocardial infarction, or electrocardiographic ST elevation. A significant increase in BNP levels was observed in the UA and NSTEMI groups compared with the stable angina group (stable angina 31.3 pg/ml, UA 147.3 pg/ml, NSTEMI, 165.8 pg/ml, p <0.01), and no differences were found between the UA and NSTEMI groups. Analysis of BNP in relation to the number of involved vessels showed significantly higher BNP levels in patients with 3- than in those with 1- or 2-vessel disease (1 to 45.2, 2 to 127.3, and 3 to 220.8 pg/ml, respectively, p <0.05 and p <0.0001, 3 vs 1- and 2-vessel disease, p = 0.01, respectively). Patients with left anterior descending stenosis had higher BNP levels than those with stenosis in other areas (150.8 vs 52.2 pg/ml, p <0.01). In conclusion, circulating BNP levels appeared elevated in patients with acute coronary syndromes with diffuse coronary involvement, even in the absence of systolic dysfunction or heart failure. BNP was also associated with multivessel disease and left anterior descending involvement. PMID- 16360360 TI - The effects of atorvastatin (10 mg) on systemic inflammation in heart failure. AB - In observational studies, statins are associated with lower mortality in patients with heart failure (HF), including those with nonischemic HF. Such benefits could be related to anti-inflammatory effects; however, the effects of statins on systemic inflammation in HF are not well-established. We conducted a 16-week, single-center, randomized, double-blind, placebo-controlled, crossover clinical trial of the effects of atorvastatin 10 mg/day on concentrations of systemic inflammatory markers in 22 patients with HF (including 20 with nonischemic HF) with New York Heart Association class II or III symptoms and left ventricular ejection fraction of <40%. The absolute and percentage of changes in inflammatory marker levels were evaluated using analysis of variance. Statin treatment reduced the concentrations of soluble tumor necrosis factor receptor-1 by 132 pg/ml (p = 0.04) and 8% (p = 0.056), C-reactive protein by 1.6 mg/L (p = 0.006) and 37% (p = 0.0002), and, after adjustment for treatment order, endothelin-1 by 0.21 pg/ml (p = 0.007) and 17% (p = 0.01). In post hoc analyses, the reduction in tumor necrosis factor receptor-1 levels was highest among patients with elevated levels at baseline (at or higher than the median of 1,055 pg/ml, p interaction = 0.001), among whom statin therapy reduced the levels by 306 pg/ml (p <0.001) and 22% (p <0.001). Statin treatment did not significantly affect the levels of other inflammatory markers, including interleukin-6 and brain natriuretic peptide. In conclusion, short-term atorvastatin therapy reduced the levels of several important inflammatory markers in patients with HF. PMID- 16360362 TI - Natural history of valvular regurgitation using side-by-side echocardiographic analysis in anorexigen-treated subjects. AB - In 1997, the US Food and Drug Administration (FDA) reported valvular regurgitation (aortic regurgitation [AR] mild or greater and/or mitral regurgitation [MR] moderate or greater) in 32.8% of patients treated with anorexigens. This study sought to determine the natural history of valvular regurgitation in patients who were included in the FDA's original report. This was a nonrandomized, natural history study of these subjects. Cohort I consisted of 155 of 186 patients (83%) who were reported to the FDA. Cohort II consisted of a similar group of 311 patients who were not included. Initial echocardiograms were obtained for medical indications before the study initiation, and study echocardiograms were standardized and obtained at 3-month intervals. Level III echocardiographers performed blinded, side-by-side comparison of the first and last study echocardiograms at a core laboratory. Cohorts I and II consisted of predominantly female, middle-aged, obese subjects. At enrollment, patients in cohorts I and II had mild or greater AR (17.9%, 13.5%) and moderate or greater MR (3.3%, 4.0%), respectively, which was less than in the FDA's report. At follow up, there were no statistically significant changes in the degree of valvular regurgitation in either cohort when the first and last study echocardiograms were read side by side. In conclusion, this natural history follow-up study demonstrates that MR and AR in long-term anorexigen-treated patients was less than reported to the FDA, did not progress, and remained echocardiographically stable during the 10-month duration of the study. PMID- 16360363 TI - Inflammatory markers and serum lipids in idiopathic dilated cardiomyopathy. AB - Coronary microcirculation is impaired in idiopathic dilated cardiomyopathy (IDC), possibly because of endothelial dysfunction. High-density lipoproteins (HDLs) have the potential to regulate endothelial function and modulate inflammation and the innate immune response. This study investigated whether reduced HDLs, concomitantly with the activation of inflammation, are associated with IDC. Fifty five patients with IDC, without evidence of other organ or systemic, chronic, or recurrent diseases, were compared with 55 healthy controls for HDLs and complete lipid profiles, C-reactive protein, C3 and C4 complement fractions, soluble intercellular adhesion molecule-1 and soluble endothelial leukocyte adhesion molecule-1, haptoglobin, and ceruloplasmin. Patients with IDC differed from controls, with lower HDL levels, lower apolipoprotein A-I and A-II levels, and higher triglyceride levels, but not on total and low-density lipoprotein cholesterol, apolipoprotein B, or lipoprotein(a). In addition, all measured inflammation markers were significantly greater in patients with IDC than in controls and were negatively correlated with HDLs. A strong and independent association with IDC was found for age, soluble intercellular adhesion molecule 1, and HDLs that, when categorized as <40 or >40 mg/dl, showed the strongest association (prevalence odds ratio 0.10, p <0.0005) with the disease. In conclusion, the data here reported on reduced HDLs and increased endothelial inflammatory activation and the linear negative correlation between HDLs and inflammation markers, particularly soluble intercellular adhesion molecule-1, could suggest a role for HDLs in the endothelial-microvascular dysfunction seen in IDC. PMID- 16360364 TI - Relation of systemic ventricular function quantified by myocardial performance index (Tei) to cardiopulmonary exercise capacity in adults after Mustard procedure for transposition of the great arteries. AB - After Mustard repair for transposition of the great arteries, the right ventricle serves as a chronically overloaded systemic ventricle (SV). Thus, during long term follow-up, dysfunction of the right ventricle with consecutive heart failure (HF) is not unusual. Early signs and symptoms of HF are depressed parameters of right ventricular (RV) function at rest and exercise intolerance. It was hypothesized that the measurement of RV function parameters, N-terminal pro-brain natriuretic peptide (NT-pro-BNP), and peak oxygen uptake (VO2max) during exercise testing were suitable for the early detection of subclinical HF. In 33 asymptomatic adolescents and adults who had undergone Mustard repair, RV function was analyzed by the myocardial performance index (Tei index). NT-pro-BNP and VO2max were also determined. The corresponding data from 52 patients operated on for left-to-right shunt defects without residual lesions served as reference data. In patients who underwent the Mustard procedure, the Tei index and NT-pro BNP were elevated (mean Tei index of the SV 0.63 +/- 0.17 vs 0.34 +/- 0.05, p <0.002; mean NT-pro-BNP 240 +/- 230 vs 57 +/- 39 pg/ml, p <0.0001), and VO2max was reduced (27 +/- 6 vs 32 +/- 6 ml x kg(-1) x min(-1), p <0.002). A good correlation was found between the Tei index and VO2max (r = -0.83, p <0.0001). In conclusion, RV function is depressed in most patients with Mustard repair. Ventricular dysfunction in such asymptomatic or minimally symptomatic patients can be detected by measurement of the Tei index, NT-pro-BNP, and VO2max. These parameters are simple and reliable screening methods to stratify patients with impaired cardiac dysfunction before they become symptomatic. PMID- 16360365 TI - Cardiac magnetic resonance imaging for accurate diagnosis of aortic arch anomalies in patients with 22q11.2 deletion. AB - This study demonstrates, in patients with 22q11.2 deletion, the wide variety of complex aortic arch anomalies that can be accurately defined by cardiac magnetic resonance imaging (MRI) compared with echocardiography. From May 1999 to August 2003, 17 of 68 patients (25%) with 22q11.2 deletion referred for cardiac evaluation required cardiac MRI, after echocardiography, to clarify aortic arch anatomy. Images of cardiac anatomy were obtained using steady-state free precession, half-Fourier acquired single turbo spin-echo dark blood sequences and gadolinium for 3-dimensional reconstruction. All MRI findings were abnormal, and in 16 of 17 cases, echocardiography was unable to define aortic arch anomalies correctly compared with MRI. PMID- 16360366 TI - Venous thromboembolism in patients with ischemic and hemorrhagic stroke. AB - The rates of pulmonary embolism (PE), deep venous thrombosis (DVT), and their combination, venous thromboembolism (VTE), in hospitalized patients with stroke from 1979 to 2003 were determined from the National Hospital Discharge Survey. Of 14,109,000 patients hospitalized with ischemic stroke, PE occurred in 72,000 (0.51%), DVT in 104,000 (0.74%), and VTE in 165,000 (1.17%). Of 1,606,000 patients hospitalized with hemorrhagic stroke, rates were higher: PE occurred in 11,000 (0.68%), DVT in 22,000 (1.37%), and VTE in 31,000 (1.93%). The rates of VTE with ischemic stroke and with hemorrhagic stroke did not change over the 25 year period of observation. PMID- 16360368 TI - Tissue Doppler velocities of the right and left ventricles and their association with C-reactive protein and homocysteine levels in Behcet's disease. AB - In the present study, we aimed to investigate whether tissue Doppler imaging could provide more information than standard echocardiography about left and right ventricular functions and their relations to homocysteine and C-reactive protein (CRP) levels in the course of Behcet's disease (BD). In the right ventricle, patients with BD had lower early diastolic tricuspid annular velocity and early diastolic/late diastolic tricuspid annular velocity ratios than the control group. Mitral and tricuspid annular systolic velocities did not significantly differ; however, diastolic velocities were correlated with duration of the disease. In addition, early/late diastolic mitral annular velocity ratio and early/late diastolic tricuspid annular velocity ratios showed an inverse relation with ocular involvement, serum homocysteine, and CRP levels in patients with BD. PMID- 16360367 TI - Implications of periaortic hematoma in patients with acute aortic dissection (from the International Registry of Acute Aortic Dissection). AB - The clinical profiles, presentation, and outcomes of patients with acute aortic dissections and associated periaortic hematomas on aortic imaging have not been described in a large cohort. This study sought to assess the prognostic implications of periaortic hematomas in patients with aortic dissections and to identify factors associated with in-hospital mortality in patients with periaortic hematomas. The study population was 971 patients with acute aortic dissections enrolled in the International Registry of Acute Aortic Dissection with available imaging data on presentation with the presence or absence of periaortic hematomas. Patients with periaortic hematomas (n = 227, 23.4%) were more likely to be women, to have a history of hypertension and atherosclerosis, and to present early to the hospital. At presentation, they had greater frequencies of shock, cardiac tamponade, coma, and/or altered consciousness. Clinical outcomes were significantly worse in patients with periaortic hematomas, including significantly greater mortality (33% vs 20.3%, p <0.001). A multivariate model demonstrated periaortic hematomas to be an independent predictor of mortality in patients with aortic dissections (odds ratio 1.71, 95% confidence interval 1.15 to 2.54, p = 0.007). In conclusion, this study provides insight into the profiles, presentation, and outcomes of patients with periaortic hematomas and acute aortic dissections. The early identification and aggressive management of patients with periaortic hematomas may potentially improve clinical outcomes. PMID- 16360369 TI - Impact of body mass index on outcome in patients undergoing carotid stenting. AB - The outcomes of patients who underwent carotid artery interventions were evaluated by body mass index (BMI). Although there was no association between BMI and short-term outcomes, patients with small BMIs were susceptible to an exaggerated long-term mortality hazard. PMID- 16360370 TI - Comparison of high-frequency two-dimensional transthoracic echocardiography versus intravascular ultrasound for evaluation of the left anterior descending coronary artery. AB - The purpose of this study was to evaluate the feasibility of high-frequency transthoracic echocardiography for measuring the wall thickness and luminal area of the left anterior descending coronary artery (LAD). Fifteen patients underwent simultaneous high-frequency transthoracic echocardiography and intravascular ultrasound (IVUS) examinations. There were good agreements for wall thickness (0.38 +/- 0.05 vs 0.38 +/- 0.06 mm, p = 0.0004) and luminal area (3.24 +/- 1.05 vs 3.32 +/- 1.34 mm2, p <0.0001) between high-frequency transthoracic echocardiography and IVUS measurements. High-frequency transthoracic echocardiography was reliable in the measurement of the wall thickness and luminal area of the LAD. PMID- 16360371 TI - Saving money and investing money: advice from a cardiologist. PMID- 16360372 TI - Role of calcium in the aortic atherosclerosis/stroke relation. PMID- 16360373 TI - Sexual dysfunction and cardiac risk (the second Princeton Concensus Conference). PMID- 16360374 TI - Acute myocardial infarction and angiographically normal coronary arteries. PMID- 16360375 TI - Carbohydrate Research. Foreword. PMID- 16360376 TI - Hypertension: a review and rationale of treatment. PMID- 16360378 TI - The prophylactic efficacy of rifampicin-soaked graft in combination with systemic vancomycin in the prevention of prosthetic vascular graft infection: an experimental study. AB - OBJECTIVE: To investigate the prophylactic efficacy of systemic, topical, or combined antibiotic usage in the prevention of late prosthetic vascular graft infection caused by methicillin-resistant Staphylococcus epidermidis (MRSE) and the differential adherence of S. epidermidis to Dacron and ePTFE grafts in a rat model. MATERIALS AND METHODS: Graft infections were established in the back subcutaneous tissue of 120 adult male Wistar rats by implantation of 1-cm(2) Dacron/ePTFE prosthesis followed by topical inoculation with 2 x 10(7) CFU of clinical isolate of MRSE. Each of the series included one group with no graft contamination and no antibiotic prophylaxis (uncontaminated control), one contaminated group that did not receive any antibiotic prophylaxis (untreated control), one contaminated group in which perioperative intraperitoneal prophylaxis with vancomycin (10 mg/kg) was administered, two contaminated groups that received rifampicin-soaked (5 mg/1 ml) or vancomycin-soaked (1 mg/1 ml) grafts, and one contaminated group that received a combination of rifampicin soaked (5 mg/1 ml) graft with perioperative intraperitoneal vancomycin prophylaxis (10 mg/kg). The grafts were removed sterilely 7 days after implantation and evaluated by using sonication and quantitative blood agar culture. RESULTS: MRSE had significantly greater adherence to Dacron than ePTFE grafts in the untreated contaminated groups (P < 0.001). Rifampicin had better efficacy than vancomycin in topical application, but the difference was not statistically significant (P > 0.05). Intraperitoneal vancomycin showed a significantly higher efficacy than topical vancomycin or rifampicin (P < 0.001). The best results were provided by a combination of intraperitoneal vancomycin in rifampicin-soaked graft groups (P < 0.001). CONCLUSIONS: The combination of rifampicin and intraperitoneal vancomycin seems to be the best choice for the prophylaxis of late prosthetic vascular graft infections caused by MRSE. PMID- 16360379 TI - Presidential address: A perspective on the future of endocrine surgery- challenges and opportunities in a time of rapid transformation. PMID- 16360380 TI - Carcinoid tumors of the duodenum. AB - BACKGROUND: Carcinoid tumors of the duodenum are rare, and their natural history has not been defined. Consequently, there is no consensus on the optimal extent of surgical treatment. METHODS: The authors reviewed the records of all patients with primary carcinoid tumors of the duodenum treated at their institution from 1969 through 2004. Patients with primary periampullary tumors and gastrinomas were excluded. RESULTS: Twenty-four patients had a pathologic diagnosis of duodenal carcinoid tumor. The majority (89%) of tumors measured less than 2 cm in diameter, and most (85%) were limited to the mucosa or submucosa. Lymph node metastases were identified in the surgical specimen in 7 (54%) of 13 patients in whom lymph nodes were examined, including 2 patients with tumors smaller than 1 cm and limited to the submucosa. At a mean follow-up of 46 months, the disease specific survival rate was 100%, and only 2 patients have had recurrences in regional lymph nodes. No patient has had distant metastases or the carcinoid syndrome. CONCLUSIONS: Carcinoid tumors of the duodenum are indolent. The presence of regional lymph node metastases cannot be predicted reliably on the basis of tumor size or depth of invasion, and their impact on survival is uncertain. PMID- 16360381 TI - Valproic acid inhibits growth, induces apoptosis, and modulates apoptosis regulatory and differentiation gene expression in human thyroid cancer cells. AB - BACKGROUND: Among the most promising new therapies for thyroid cancer are the histone deacetylase inhibitors. Valproic acid (VA) is an anticonvulsant that inhibits histone deacetylase activity at nontoxic concentrations. We hypothesized that VA would have antineoplastic effects on human thyroid cancer cells. METHODS: We treated 1 papillary and 3 follicular thyroid cancer cell lines with VA (0.5-2 mmol/L) for 24 to 72 hours. Cell proliferation was measured with a cell proliferation assay kit. Annexin V-fluorescein isothiocyanate was used to quantitate cells that were undergoing apoptosis. Quantitative polymerase chain reaction was used to measure expression of apoptosis-regulatory and differentiation genes. RESULTS: VA inhibited growth in all cell lines by 26% to 59% at 48 hours and up to 77% at 72 hours. Nineteen percent to 30% of VA-treated cells underwent apoptosis, compared with 4% to 8% of the control cells. Expression of pro survival genes bcl-2 and bcl-xl was down-regulated by 10% to 60%; expression of the proapoptosis gene bax was up-regulated by 23% to 85%. Sodium-iodide symporter and thyroglobulin messenger RNA expression were up regulated by 93% to 370% in follicular cell lines but remained unchanged in the papillary cell line. CONCLUSION: VA inhibits growth, induces apoptosis, and modulates apoptosis-regulatory and differentiation gene expression in thyroid cancer cells. These findings suggest that VA may be useful clinically for patients with thyroid cancers of follicular cell origin. PMID- 16360382 TI - Selective arterial chemoembolization for hepatic metastases from medullary thyroid carcinoma. AB - BACKGROUND: Hepatic metastases from medullary thyroid carcinoma (MTC) may impair quality of life by hypercalcitonemia-associated diarrhea and pain. In this prospective study, the effect of selective arterial chemoembolization (SACE) was evaluated. METHODS: Eleven patients with hepatic metastases from MTC received 1 to 9 courses of SACE using epirubicine. Symptomatic, biochemical, and morphologic responses on SACE were recorded. RESULTS: Symptomatic response was observed in all symptomatic patients. However, biochemical and radiologic response occurred only in 6 patients. Liver function was not affected by SACE. One patient with unexpected concurrent pheochromocytoma metastases died after the first course. Development of side effects in the course was observed in 8 patients but were only World Health Organization grade 1. Patients' satisfaction with SACE was excellent. Long-term follow-up found 7 patients alive (1-72 months). Three patients died with tumor 6, 12, and 24 months after SACE, respectively. CONCLUSION: SACE provided good symptom palliation for the majority of patients with hepatic metastases from MTC. However, transient remission or stabilization of hepatic metastases resulted in only 60%. Further studies using a randomized protocol are required. PMID- 16360383 TI - MUC1 plays a role in tumor maintenance in aggressive thyroid carcinomas. AB - BACKGROUND: We recently identified MUC1 as a target driving selection for 1q21 amplification and validated it as an independent marker of aggressive behavior in thyroid cancer (TC). The aims of this study were to determine whether TC cell lines retain MUC1 expression patterns that are seen in primary tumors, assess the role of MUC1 in tumor maintenance, and develop a virally delivered anti-MUC1 RNA interference (RNAi) that is effective in decreasing MUC1 expression in vitro. METHODS: Fifteen TC cell lines were screened for MUC1 protein expression. Cell lines with varying MUC1 protein levels were treated with anti-MUC1 monoclonal antibody to assess cell viability. A recombinant retroviral short hairpin RNAi delivery system against MUC1 was developed. Efficacy and optimal dosing of short hairpin RNA against MUC1 was determined. RESULTS: MUC1 expression patterns in TC cell lines were found to be similar to that seen in primary tumors. Treatment with anti-MUC1 antibody resulted in a significant decrease in cell viability in MUC1 over-expressing cell lines. MUC1-779 RNAi construct showed excellent infection efficiency and reproducible silencing. CONCLUSION: These data offer functional evidence that implicates MUC1 over-expression as a key molecular event in the pathogenesis of aggressive TC. Retrovirally delivered anti-MUC1 RNAi is effective in silencing MUC1 and merits further investigation to establish therapeutic efficacy and safety in anticipation of potential clinical application. PMID- 16360384 TI - A prospective evaluation of laparoscopic exploration with intraoperative ultrasound as a technique for localizing sporadic insulinomas. AB - BACKGROUND: Preoperative imaging studies localize insulinomas in less than 50% of patients. Arteriography with calcium stimulation and venous sampling (ASVS) regionalizes greater than 90% of insulinomas but requires specialized expertise and an invasive procedure. This prospective study evaluated laparoscopic exploration with IOUS compared with the other localization procedures in patients with a sporadic insulinoma. METHODS: Between March 2001 and October 2004, 14 patients (7 women and 7 men; mean age, 53) with an insulinoma were enrolled in an IRB-approved protocol. Computed tomography, magnetic resonance imaging, ultrasound scan, and arteriography with calcium stimulation and venous sampling were performed preoperatively. A surgeon, blinded to the results of the localizing studies, performed a laparoscopic exploration with intraoperative ultrasound (IOUS). At the completion of the exploration, the success of laparoscopy for localization was scored, and the tumor was resected. RESULTS: Twelve of 14 tumors were localized successfully before laparoscopy (noninvasive, 7 of 14; invasive, 11 of 14). Laparoscopic IOUS localized successfully 12 of 14 tumors. All lesions were resected, and all patients were cured (median follow-up, 36 months). CONCLUSION: Laparoscopic IOUS identified 86% of tumors. The authors consider laparoscopic IOUS to be equivalent to ASVS in localizing insulinomas. Further study is therefore warranted to determine the role of laparoscopy with IOUS in the localization and treatment algorithm for patients with sporadic insulinoma. PMID- 16360385 TI - Clinical outcomes of laparoscopic adrenalectomy for lateralizing nodular hyperplasia. AB - BACKGROUND: Nodular adrenal hyperplasia (NAH) may mimic the biochemical characteristics of an aldosterone-producing adenoma. The authors evaluated the outcomes of unilateral laparoscopic adrenalectomy in the setting of lateralizing aldosterone hypersecretion by NAH. METHODS: Retrospective review of consecutive patients who underwent a laparoscopic adrenalectomy for primary hyperaldosteronism owing to NAH was performed. Patient demographics, perioperative symptoms, medications, radiographic findings, and serum chemistries were analyzed. Response to operation was classified according to postoperative control of hypertension and hypokalemia as resolved, improved, or refractory. RESULTS: From January 1999 to October 2004, 15 patients underwent a laparoscopic unilateral adrenalectomy for hyperaldosteronism owing to lateralizing NAH. Nine (60%) patients presented with > or =5 years of hypertension, including 8 (53%) patients with labile or malignant hypertension. Ten (67%) patients had hypokalemia. Abdominal imaging results were normal in 9 (60%) patients. All patients underwent adrenal venous sampling (94% successfully), which revealed an average adjusted aldosterone ratio of 17.6 (range, 1.2 to 75.9). At a mean follow up of 26 (range, 4 to 58) months, hypertension had resolved in 4 (27%), improved in 8 (53%), and was refractory in 3 (20%) patients. Hypokalemia resolved in all patients. There were no complications, conversions, or mortalities. CONCLUSION: This series shows that unilateral adrenalectomy for lateralizing NAH results in eradication of hypokalemia and resolution or significant improvement in hypertension in 80% of patients at long-term follow-up. When lateralization of aldosterone production is noted, laparoscopic adrenalectomy provides significant clinical improvement even in patients with a pathologic diagnosis of NAH. PMID- 16360386 TI - 25-hydroxyvitamin D deficiency is a risk factor for symptoms of postoperative hypocalcemia and secondary hyperparathyroidism after minimally invasive parathyroidectomy. AB - BACKGROUND: Patients with primary hyperparathyroidism who undergo minimally invasive parathyroidectomy (MIP) may have postoperative symptoms of hypocalcemia or secondary hyperparathyroidism. This study sought to identify factors predictive of these events. METHODS: Between 1998 and 2004, 190 patients with primary hyperparathyroidism underwent MIP with excision of a single adenoma. Age, gender, race, prior head and neck surgery, use of preoperative thyroid hormone or calcium-channel blockers, preoperative levels of calcium, 25-hydroxyvitamin D (25[OH]D) and intact parathyroid hormone (iPTH), the presence of osteopenia or osteoporosis, intraoperative iPTH levels, and adenoma weight were evaluated by univariate analysis as predictors of postoperative symptoms of hypocalcemia and secondary hyperparathyroidism. RESULTS: None of the following were predictors of postoperative symptoms of hypocalcemia: age, gender, race, prior head and neck surgery, preoperative medications, preoperative calcium and iPTH levels, osteopenia or osteoporosis, intraoperative iPTH levels, or adenoma weight. However, patients with postoperative symptoms of hypocalcemia had significantly lower preoperative 25[OH]D levels (P = .01). Further, higher preoperative iPTH levels (P < .01) and lower preoperative 25[OH]D levels (P = .05) were associated with secondary hyperparathyroidism postoperatively. CONCLUSIONS: A low preoperative 25[OH]D level is associated with postoperative symptoms of hypocalcemia and secondary hyperparathyroidism in patients undergoing MIP. One might consider instituting empiric calcium supplementation postoperatively in patients with low 25[OH]D levels. PMID- 16360387 TI - Revisiting lithium-associated hyperparathyroidism in the era of intraoperative parathyroid hormone monitoring. AB - BACKGROUND: Chronic lithium therapy may cause hyperparathyroidism (HPT). The utility of intraoperative parathyroid hormone monitoring (IOPTH) in these patients is unknown. The authors' hypothesis was that multiglandular disease is more common in these patients, and the ability of IOPTH to predict cure may be limited. METHODS: Twelve patients had HPT during chronic lithium therapy and underwent parathyroidectomy with IOPTH. Criteria for curative resection were a decrease > or =50% from baseline and into the normal range. Calcium and PTH levels were measured during follow-up. RESULTS: Preoperatively, mean calcium was 11.0 +/- 0.1 mg/dL, and PTH was 116 +/- 14 pg/mL. Fifty percent of patients had multiglandular disease confirmed by IOPTH levels. Mean IOPTH decrease from baseline was 74 +/- 4%. Although 10 of 12 patients met criteria for curative resection, only 8 remain normocalcemic. The 2 patients who did not meet criteria remain normocalcemic. Mean postoperative calcium for all patients was 9.5 +/- 0.2 mg/dL. Of the 10 normocalcemic patients, 4 also have hyperparathormonemia (mean PTH, 119 +/- 19 pg/mL). CONCLUSIONS: The incidence of multiglandular disease in HPT after chronic lithium exposure is higher than standard HPT. The ability of IOPTH to predict durable normocalcemia is limited. Bilateral neck exploration should be considered for these patients regardless of whether IOPTH monitoring is used. PMID- 16360388 TI - Long-term functionality of cryopreserved parathyroid autografts: a 13-year prospective analysis. AB - BACKGROUND: The functional results of cryopreserved heterotopic parathyroid autotransplantation (CHPA) are not well defined. The authors evaluated the outcomes of delayed CHPA for the treatment of surgically induced hypoparathyroidism. METHODS: Since November 1991, 448 parathyroid samples from 436 patients were cryopreserved at our institution. Of these, 29 patients underwent 34 CHPA procedures, with placement of 20 to 25 pieces of parathyroid tissue (approximately 50 to 75 mg) into the forearm. Outcomes were determined based on peripheral parathyroid hormone (PTH) levels and, where available, PTH gradients between grafted and nongrafted arms. Graft function results were defined as completely functional (patients with normal PTH and calcium levels off all calcium/vitamin D supplementation), partially functional (normal PTH levels and mild hypocalcemia on calcium supplementation), or nonfunctional (low PTH levels and dependent on calcium/vitamin D supplementation). RESULTS: Of the 29 patients with CHPA, prospective data were available for 26 patients undergoing 30 CHPA procedures (9 patients with MEN 1, 4 with MEN 2A, 1 with MEN 2B, and 12 with sporadic hyperparathyroidism). The mean follow-up interval was 2 years. Twelve of 26 patients (46%) had completely functional grafts, 6 patients (23%) had partially functional grafts, and the remaining 8 patients (31%) had nonfunctional grafts. No patient with CHPA had graft-dependent recurrent hyperparathyroidism. Of the 14 patients (15 autografts) with MEN, 7 patients (50%) had fully functional grafts, and 2 patients (14%) had partially functional grafts. The mean cryopreservation period was 7.9 months (range, 1 week to 22 months) for functional autografts and 15.3 months (range, 2 weeks to 106 months) for nonfunctional autografts (P < .01). CONCLUSIONS: Based on these data and those in previous studies, approximately 60% of delayed, cryopreserved parathyroid autografts are functional. In this study 40% autografts (46% of patients) achieved full competency off supplements. Some patients have evidence of graft function with normal PTH levels but are not normocalcemic. Results were similar for patients with MEN and nonhereditary hyperparathyroidism. The duration of cryopreservation was a significant indicator of graft failure, and no functional autograft was observed beyond 22 months of preservation. CHPA is a useful treatment modality for patients with postoperative hypocalcemia after thyroid or parathyroid surgery, who do not respond to immediate parathyroid autotransplantation. PMID- 16360389 TI - Molecular analysis of minimally invasive follicular carcinomas by gene profiling. AB - BACKGROUND: While the majority of minimally invasive follicular thyroid carcinoma (MIFTC) behave like follicular adenomas, some recur or metastasize. These studies were conducted to determine if molecular profiling can enhance our understanding of MIFTC and to perhaps offer a better classification schema. METHODS: Microarray analysis was performed on 27 follicular neoplasms. Thirteen follicular adenomas (FAs) were compared with 7 widely invasive follicular thyroid carcinomas (WIFTCs) to generate a list of differentially expressed genes. Next, 7 MIFTCs were analyzed along with the other samples in a cluster analysis. The MIFTCs were then compared directly against both the adenoma and WIFTC groups to investigate genetic relatedness. RESULTS: FAs and WIFTCs have distinct genetic profiles, with 401 differentially expressed genes. The 7 MIFTCs were added to the analysis. Six of 7 of the MIFTCs were grouped with the adenomas, 4 of which created their own subgroup. When analyzed directly, MIFTCs had 223 differently expressed genes, compared with FAs, and 365, compared with WIFTCs. CONCLUSIONS: Molecular profiling illustrates that the majority of MIFTCs comprise a subclass of follicular neoplasms, and, while most MIFTCs are genetically similar to adenomas, our patient data suggest that these tumors may deserve greater suspicion of malignant potential. Gene profiling can provide insight into the molecular pathogenesis of MIFTC. PMID- 16360390 TI - A six-gene model for differentiating benign from malignant thyroid tumors on the basis of gene expression. AB - BACKGROUND: Thyroid nodules are common; fine-needle aspirations commonly are read as indeterminate, necessitating surgery to exclude carcinoma. We developed a 6 gene array-based predictor model to diagnose benign versus malignant thyroid lesions. In this study, we verified whether quantitative reverse transcription polymerase chain reaction (qRT-PCR) using this model reliably can differentiate benign from malignant thyroid nodules. METHODS: Molecular profiles of benign (follicular adenomas, hyperplastic nodules) and malignant tumors (papillary thyroid carcinomas, follicular variants of papillary thyroid carcinomas) were analyzed using qRT-PCR from our 6-gene model (kit, Hs.296031, Hs.24183, LSM7, SYNGR2, C21orf4). The gold standard was standard pathologic criteria. A diagnosis predictor model was built by using the training samples and was then used to predict the class of 10 additional samples analyzed as unknowns. RESULTS: Our predictor model using 47 training samples correctly predicted 9/10 unknowns. One sample diagnosed as benign by standard histologic criteria was diagnosed as malignant by our model (sensitivity 75%; specificity, 100%; positive predictive value, 100%; negative predictive value, 85.7%). CONCLUSIONS: Molecular diagnosis with our 6-gene model can differentiate between benign and malignant thyroid tumors with high sensitivity and specificity. In combination, these genetic markers may be a reliable test to preoperatively diagnose the malignant potential of thyroid nodules. PMID- 16360391 TI - Hyperthyroidism after parathyroid exploration. AB - BACKGROUND: We hypothesized that hyperthyroidism after parathyroid exploration may be an underreported phenomenon with a course more severe than recognized previously. METHODS: We examined pre- and postoperative thyroid function and outcomes in 199 consecutive patients who, since March 2000, had parathyroid exploration for primary sporadic hyperparathyroidism (HPTH). We excluded patients with prior thyroid or parathyroid surgery, preoperative thyroid medication, concurrent total thyroidectomy, or follow-up <5 months. RESULTS: Of 125 patients with normal preoperative serum thyroid-stimulating hormone levels, 39 (31.2%) were hyperthyroid postoperatively. Mean thyroid-stimulating hormone levels (mean +/- SD) dropped with operation from 2.0 +/- 1.1 microIU/mL to 1.2 +/- 1.4 microIU/mL (P < .0001). Nineteen patients (15%) reported symptoms 1 to 2 weeks after operation. The clinical course of hyperthyroidism typically was short, but 5 patients (4%) had symptomatic hyperthyroxinemia requiring medical therapy. Hyperthyroidism was independent of age, severity of HPTH, anatomic/pathologic features, operative time, and other measures of operative difficulty, but was associated with lithium therapy, bilateral exploration, and absence of concurrent thyroid lobectomy. CONCLUSIONS: Risk of hyperthyroidism may be underappreciated after routine parathyroid surgery for HPTH. Use of lithium and degree of dissection appear contributory. Patients undergoing parathyroid exploration need counseling and surveillance for hyperthyroidism, which may be reduced by minimizing the extent of parathyroid surgery. PMID- 16360392 TI - The role of intraoperative parathyroid hormone testing in patients with tertiary hyperparathyroidism after renal transplantation. AB - BACKGROUND: Intraoperative parathyroid hormone (PTH) testing has been shown to accurately define adequacy of parathyroid resection in patients with primary hyperparathyroidism (HPT) and alters the operative management in 10% to 15% of cases. However, the benefit of this technique in patients with tertiary HPT after renal transplantation undergoing parathyroidectomy is unclear. METHODS: Intraoperative PTH was measured in 32 consecutive patients undergoing parathyroidectomy for tertiary HPT after renal transplantation between March 2001 and November 2004 by using the Elecsys assay at baseline and, subsequently, 5, 10, and 15 minutes after curative resection. The outcomes of these patients were evaluated. RESULTS: All patients were cured after surgery. Of the 32 patients, 29 were found to have parathyroid hyperplasia, while 1 had a single adenoma and 2 had double adenomas. The average drop in intraoperative PTH levels after curative resection was 69 +/- 3.5% at 5 min., 77 +/- 2.3% at 10 minutes, and 83 +/- 3.4% at 15 minutes. PTH testing changed the intraoperative management in 5 (16%) patients. One patient with a single adenoma and 2 patients with double adenomas had a >50% drop at 10 minutes. after excision; therefore, the operation was terminated without further resection. Two patients did not have a >50% drop at 10 minutes after 3.5 gland resection. These patients were explored further, and additional supernumerary parathyroid glands were identified and resected. After resection of these additional glands, the PTH fell by >50%, indicating cure. CONCLUSIONS: In patients undergoing parathyroidectomy for tertiary HPT after renal transplantation, a decrease in intraoperative PTH levels >50% at 10 minutes after completion of the operation indicated adequate resection. Furthermore, intraoperative PTH testing altered the operative management in 16% of patients. Therefore, similar to its role in patients with primary HPT, intraoperative PTH testing appears to play an equally important role in the management of patients with tertiary HPT undergoing parathyroidectomy. PMID- 16360393 TI - Radioactive iodine offers survival improvement in patients with follicular carcinoma of the thyroid. AB - BACKGROUND: The use of radioactive iodine (RAI) in patients with follicular thyroid carcinoma is well established. How its use affects patient outcome and which patients benefit is understood poorly. This study seeks to determine how RAI influences survival and to delineate the populations that are impacted most. METHODS: The Surveillance, Epidemiology, and End Results database is a sample of approximately 14% of the US population. It was used to identify patients with follicular thyroid carcinomas and the treatment that they received. Factors such as the presence of lymph node and distant metastases, age, and tumor size were included for analysis. RESULTS: A total of 4317 patients were identified with follicular thyroid carcinoma. Of these, the records of 2112 patients who were entered in the study after 1988 contained the required data and were included for analysis. Median follow-up time was 95 months. Factors that were associated with increased risk of death included distant metastatic disease, cervical lymph node disease, and the lack of RAI use. Protective factors were tumor size of <2 cm and age of <45 years. Some patients with a greater number of risk factors benefited from RAI. CONCLUSION: RAI provides survival benefit to some patients with follicular carcinoma of the thyroid. The greatest improvements were seen in those patients with locoregional or distant disease spread. PMID- 16360394 TI - Laparoscopic resection of adrenal cortical carcinoma: a cautionary note. AB - BACKGROUND: While laparoscopic removal of small, benign, functioning adrenal tumors is accepted, laparoscopic resection of adrenal tumors that may be adrenal cortical carcinoma (ACC) remains controversial. METHODS: The records of all patients with ACC evaluated at a single institution from 1991 through 2004 were reviewed retrospectively. RESULTS: Among 170 patients with ACC, 153 patients underwent open anterior adrenalectomy, 6 underwent laparoscopic adrenalectomy, 1 was treated via an open flank approach, and 10 had no operation. At a median follow-up of 28 months, 115 (86%) of 133 patients who had undergone open anterior resection of primary ACC had had a recurrence. Local recurrence and peritoneal carcinomatosis were components of initial failure in 46 (35%) and 11 patients (8%), respectively. In contrast, all 6 patients who underwent laparoscopic resection of ACC had recurrences, and peritoneal carcinomatosis was a component of initial failure in 5 (83%) of them (open vs laparoscopic resection, Fisher exact test P = .0001). CONCLUSIONS: Laparoscopic resection of ACC is associated with a high risk of peritoneal carcinomatosis. Open adrenalectomy remains the standard of care for patients presenting with an adrenal cortical tumor for which ACC is in the differential diagnosis. PMID- 16360395 TI - Expression profiling of adrenocortical neoplasms suggests a molecular signature of malignancy. AB - BACKGROUND: Distinguishing between adrenocortical adenomas and carcinomas is often difficult. Our aim was to investigate the differences in transcriptional profiles between benign and malignant adrenocortical neoplasms using complementary DNA microarray techniques. METHODS: We studied 7 patients with adrenocortical carcinomas and 13 with adenomas. Histopathology was reviewed in all patients; clinical follow-up was at least 1 year. Hybridizations were performed in duplicate against RNA reference. Expression levels were analyzed in the R environment for statistical computing with the use of aroma, limma, statistics, and class packages. RESULTS: Transcriptional profiles were homogeneous among adenomas, while carcinomas were much more heterogeneous. Hierarchical clustering and self-organizing maps could separate clearly carcinomas from adenomas. Among genes that were most significantly upregulated in carcinomas were 2 ubiquitin-related genes (USP4 and UFD1L) and several insulinlike growth factor-related genes (IGF2, IGF2R, IGFBP3 and IGFBP6). Among genes that were most significantly downregulated in carcinomas were a cytokine gene (CXCL10), several genes related to cell metabolism (RARRES2, ALDH1A1, CYBRD1 and GSTA4), and the cadherin 2 gene (CDH2). CONCLUSIONS: Through the use of cDNA arrays, adrenocortical adenomas and carcinomas appear to be clearly distinguishable on the basis of their specific molecular signature. The biologic importance of the up- and downregulated genes is yet to be determined. PMID- 16360396 TI - Nodal yield, morbidity, and recurrence after central neck dissection for papillary thyroid carcinoma. AB - BACKGROUND: The role of central neck dissection (CND) in differentiated thyroid cancer remains controversial. This study aims at elucidating the potential benefits and drawbacks of CND associated to total thyroidectomy in papillary cancer. METHODS: Protocols of patients undergoing total thyroidectomy and CND for papillary cancer were reviewed. The following data were recorded: macroscopic appearance of central nodes; nodes obtained at operation; number of metastatic nodes and parathyroid glands incidentally resected; metastases, age, completeness, invasiveness, size score; postoperative s-Ca; complications; and recurrences. Differences between therapeutic (gross nodal involvement) and prophylactic (no apparent node involvement) CNDs were studied. RESULTS: Forty three patients (mean age, 52 +/- 17 years) were studied. A mean of 8.4 +/- 6.6 nodes were resected per patient. A 60% prevalence (26/43) of presence of nodal involvement (N+) was found with no difference between low- and high-risk patients. Twenty-five (60%) patients developed transient hypocalcemia, which was associated with incidental parathyroidectomy, number of nodes resected, and thymectomy. Two patients (4.6%) developed permanent hypoparathyroidism and 3 (7%), transient vocal cord paralysis. Parathyroid glands were found in 19% of the specimens. At follow-up, there were no central neck recurrences, but 5 patients developed lateral recurrences despite treatment with I(131). All 5 patients had had therapeutic CND with 6 or more metastatic nodes obtained in the CND specimen. No lateral neck recurrences were observed after prophylactic CND or in patients with < 6 nodes involved. CONCLUSIONS: CND prevents central neck recurrences. Morbidity of bilateral CND is significant, and its systematic implementation in the absence on gross nodal involvement requires reassessment. PMID- 16360397 TI - Diagnostic and prognostic value of angiogenesis-modulating genes in malignant thyroid neoplasms. AB - BACKGROUND: Angiogenesis is an essential biologic event in the pathogenesis of human malignancies. We postulated that expression analysis of genes that modulate angiogenesis would identify differentially expressed genes that would help to distinguish benign from malignant thyroid neoplasms and serve as markers of aggressive differentiated thyroid cancer. METHODS: A complementary DNA (cDNA) array with 96 genes that modulate angiogenesis was used to identify differentially expressed genes (2-fold higher or lower) in malignant versus benign thyroid neoplasms. Real-time quantitative polymerase chain reaction was used to confirm cDNA array expression data in 123 patients (4 normal thyroid, 26 hyperplastic nodules, 27 follicular adenomas, 23 follicular cancers, 18 follicular variant of papillary cancers, 25 papillary cancers). RESULTS: Twenty two genes were upregulated in malignant thyroid neoplasms by cDNA array analysis, but only 13 genes had higher messenger RNA (mRNA) expression levels in malignant than in benign thyroid neoplasms by real-time quantitative polymerase chain reaction (P < or = .04). Of the 13 differentially expressed genes, the combined use of angiopoietin 2 (ANGPT2) and tissue inhibitor of metalloproteinase 1 (TIMP1) mRNA expression levels was best for distinguishing malignant from benign thyroid neoplasms, with a sensitivity of 90%, specificity of 85%, positive predictive value of 75%, and negative predictive value of 94%. Epidermal growth factor receptor and ephrin B2 mRNA expression was elevated in higher TNM stage neoplasms and in patients with high-risk AMES (Age, distant Metastasis, Extrathyroidal invasion, and tumor Size) differentiated thyroid cancers (P < or = .005). CONCLUSIONS: Angiopoietin 2 and tissue inhibitor of metalloproteinase 1 are diagnostic markers of malignant thyroid nodules and could improve the diagnostic accuracy of FNA biopsy. Epidermal growth factor receptor and ephrin B2 are markers of aggressive differentiated thyroid cancer. PMID- 16360398 TI - Surgery for hyperparathyroidism: does morphology or function matter most? AB - BACKGROUND: With minimally invasive parathyroidectomy (MIP) not all enlarged parathyroid glands are necessarily removed, and intraoperative measurement of parathyroid hormone levels (IO-PTH) does not necessarily predict multiple enlarged glands. The aim of this study was to compare morphology with function, using Ca(2+)-regulated PTH secretion. METHODS: PTH secretion was determined by perifusion: (1) cells from 12 normal parathyroids were compared with 14 parathyroid adenomas; (2) functional characteristics (PTH secretion, sestamibi uptake, IO-PTH decrease) were correlated with morphologic characteristics; (3) PTH secretion as a predictor of IO-PTH decrease was determined in 7 patients with 2 enlarged parathyroids. RESULTS: (1) There were significant differences between normal and pathological parathyroid cells consistent with reduced sensitivity to Ca(2+). Maximum secretion rates for normal and adenomatous cells were, respectively, 3.9 +/- 0.4 fg min(-1) cell(-1) and 2.0 +/- 0.4 fg min(-1) cell(-1) (P = .002) and minimum secretion rates, 0.7 +/- 0.1 fg min(-1) cell(-1) and 0.4 +/- 0.1 fg min(-1) cell(-1) (P = .008). However, the IC(50) value for Ca(2+) was elevated in adenomatous cells indicating an apparent loss of extracellular Ca(2+) sensitivity being 1.1 +/- 0.02 mmol/L for normal and 1.2 +/- 0.02 mmol/L for adenomatous cells (P = .02). (2) There was no overall correlation between PTH secretion and gland morphology. (3) In 5 of 7 cases, PTH secretion correctly predicted the decrease in IO-PTH. CONCLUSION: Parathyroid adenomas generally exhibit abnormal PTH secretory function; however, enlarged parathyroid glands that do not contribute to the biochemical changes of hyperparathyroidism do exist, and, in these cases, cellular secretory function is a useful predictor of IO-PTH dynamics. PMID- 16360399 TI - Parathyroidectomy improves neurocognitive deficits in patients with primary hyperparathyroidism. AB - BACKGROUND: Clinical guidelines for the treatment of primary hyperparathyroidism (pHPT) often suggest parathyroidectomy, but generally fail to consider neurocognitive and psychiatric symptoms because of the relative paucity of evidence. METHODS: In this prospective study, patients with pHPT (PTX) and benign euthyroid thyroid disease (THY) referred for operation were evaluated pre- and postoperatively with validated psychometric and neurocognitive instruments to determine whether learning, memory, or concentration improved with after parathyroidectomy. Statistical comparisons between groups were performed with univariate analysis and repeated measures of analysis of variance. RESULTS: Fifty five subjects, mean age of 54 years, were evaluated preoperatively; 41 returned postoperatively. There were no significant differences between groups by age and gender. PTXs reported more depression symptoms preoperatively (P = .04) that improved postoperatively. There were no differences between the 2 groups on verbal memory and trait anxiety. For PTXs, average preoperative serum calcium concentration (11.3 mg/dL) and serum PTH level (100 pg/mL) normalized postoperatively. Preoperatively PTXs showed greater delays in their spatial learning (P = .03). All subjects learned across the 5 trials, but PTXs were more delayed (P = .03). After operation, PTXs improved and functioned at a level equivalent to the THYs. There was an interaction between trial (neurocognitive testing), visit (pre- vs postoperative), status (PTX vs THY), and change in PTH level (P = .06), suggesting that individuals with greater change in PTH were more likely to improve in their learning efficiency postparathyroidectomy. CONCLUSIONS: PHPT may be associated with a spatial learning deficit and processing that improves after parathyroidectomy. While longer-term follow-up is necessary, neurocognitive symptoms perhaps should be considered as criteria for parathyroidectomy. PMID- 16360400 TI - Persistently elevated parathyroid hormone levels after parathyroid surgery. AB - BACKGROUND: Persistent elevation of serum parathyroid hormone (PTH), despite normocalcemia, occurs in 8% to 40% of patients after parathyroidectomy. Explanations have included hypocalcemia owing to vitamin D deficiency or bone remineralization, and persistent hyperparathyroidism. METHODS: A retrospective chart review of 816 consecutive patients who underwent surgery for primary hyperparathyroidism was conducted. RESULTS: One hundred fourteen patients (15%) had persistently elevated PTH levels (PPTH). Patients with PPTH had higher preoperative PTH levels than those with normal PTH levels postoperatively. They also had lower postoperative Ca(++) and vitamin D levels. Multiple gland enlargement was identified in fewer patients with PPTH than in those with normal postoperative PTH levels. In patients with PPTH and a postoperative Ca(++) less than 9.6 mg/dL (group I), there was a greater decrease in IOPTH, a higher initial postoperative PTH level, and a lower postoperative vitamin D level than in PPTH patients whose postoperative Ca(++) was > or =9.6 mg/dL (group II). Postoperative Ca(++) and vitamin D levels were also lower in patients whose PPTH did not ultimately resolve. Three patients in group II had recurrent disease. CONCLUSIONS: Persistent elevation of postoperative serum PTH levels in normocalcemic patients is associated with mild hypocalcemia, probably owing to vitamin D deficiency. In some patients it may also be indicative of mild persistent hyperparathyroidism. PMID- 16360401 TI - Hairy Enhancer of Split-1 (HES-1), a Notch1 effector, inhibits the growth of carcinoid tumor cells. AB - BACKGROUND: The Notch1-signaling pathway has been shown to regulate the differentiation and growth of carcinoid tumor cells. However, the molecules that mediate Notch1 signaling, as well as their potential roles in regulating the growth of carcinoid tumors, have not been characterized. We and others have shown previously that the transcription factor Hairy Enhancer of Split-1 (HES-1) is upregulated in response to Notch1 signaling, demonstrating that it is a Notch1 effector. We hypothesized that HES-1 may be the essential downstream factor in Notch1-mediated growth regulation of carcinoid tumors. METHODS: H727 carcinoid tumor cells were transduced stably with a doxycycline-inducible HES-1 construct, creating H727-HES-1 cells. H727-TRE (vector-only control) and H727-HES-1 cells were then treated with varying concentrations of doxycycline to achieve increasing levels of HES-1 protein expression. Cell proliferation was determined with the use of a cell viability assay. RESULTS: Treatment of H727-HES-1 cells with increasing dosages of doxycycline resulted in dose-dependent increases in HES-1 protein by Western blot analysis. Importantly, induction of HES-1 in carcinoid tumor cells led to suppression of tumor cellular proliferation. Moreover, the degree of carcinoid growth inhibition appeared to be proportional to the level of HES-1 induction. CONCLUSIONS: HES-1 alone can regulate the growth of carcinoid tumor cells. Furthermore, these results suggest that HES-1 may be the critical downstream effector in the Notch1-signaling pathway. PMID- 16360402 TI - Use of surgeon-performed office ultrasound and parathyroid fine needle aspiration for complex parathyroid localization. AB - BACKGROUND: This study investigates the utility of ultrasound guided parathyroid fine needle aspiration (FNA) as a localizing technique in patients with hyperparathyroidism (HPT) undergoing re-operative neck surgery or with unusually appearing or ectopically located glands. METHODS: Selected patients with HPT underwent surgeon-performed FNA with ultrasound guidance. Aspirate contents were sent for cytology and parathyroid hormone (PTH) levels. All patients subsequently underwent parathyroid exploration. RESULTS: 54 patients underwent 57 ultrasound guided parathyroid biopsies. Indications for FNA included prior parathyroid (n = 29), thyroid (n = 11), or other neck surgery (n = 2), or unusual parathyroid appearance or location (n = 12). A true positive was defined as a site where the PTH aspirate was >40 pg/mL and a hypercellular gland was removed at surgery. Based on this, there were 44 true positives, 10 true negatives, and 3 false negatives; there were no false positives. The median PTH level in positive aspirates was 11,665 pg/mL. Cytology was primarily helpful in excluding other diagnoses. CONCLUSION: Ultrasound guided FNA is a highly specific localization test for parathyroid tumors. This procedure can be successfully performed by surgeons in the office setting and is extremely valuable for directing parathyroid exploration in challenging cases. We recommend incorporating ultrasound and FNA as a pre-operative localization strategy for selected patients with HPT. PMID- 16360403 TI - The treatment and prognosis of Hurthle cell follicular thyroid carcinoma compared with its non-Hurthle cell counterpart. AB - BACKGROUND: This population study compared the treatment and prognosis of Hurthle cell follicular thyroid carcinoma (HCFC) and non-HCFC. METHODS: The Surveillance, Epidemiology and End Results database identified patients with HCFC and non-HCFC from 1988 to 1993 who were followed to 2001. Treatment of each carcinoma was compared, and the effect of prognostic factors on survival was analyzed. RESULTS: Eight hundred forty-five patients were identified; 172 patients (20%) had HCFC and 673 patients (80%) had non-HCFC. Total thyroidectomy was performed in 80% of patients with HCFC compared with 69% with non-HCFC (P = .005). Radioactive iodine was used in 33% with HCFC and 45% with non-HCFC (P = .003). The crude 10-year survival was 73% in HCFC and 83% in non-HCFC patients. Older age (> or =50 vs <50 years; hazard ratio, 6.35; 95% CI, 4.07-9.93), men (hazard ratio, 2.07; 95% CI, 1.52-2.81), larger tumor size (>5 vs < or =5 cm; hazard ratio, 2.20; 95% CI, 1.55 3.13; >10 cm vs < or =5 cm; hazard ratio, 3.28; 95% CI, 1.12-9.61), nodal metastases (hazard ratio, 3.11; 95% CI, 1.80-5.37), and distant metastases (hazard ratio, 3.91; 95% CI, 1.94-7.90) were associated with a higher mortality rate. Histologic type (non-HCFC vs HCFC; hazard ratio, 0.85; 95% CI, 0.60-1.19; P = .34), local extension, extent of thyroidectomy, and radioactive iodine use had no effect on the mortality rate. CONCLUSIONS: Histologic distinction between HCFC and non-HCFC is not as prognostically important as age, gender, and tumor stage. This study suggests that patients with HCFC should be treated the same as patients with equivalent stage non-HCFC. PMID- 16360404 TI - Are frozen sections useful and cost-effective in the era of intraoperative qPTH assays? AB - BACKGROUND: Since intraoperative quick parathormone (IOqPTH) assays are available, the role of frozen sections (FS) during parathyroid exploration has become questionable. This study compares the results of FS and IOqPTH in primary hyperparathyroidism (pHPT). METHODS: FS and IOqPTH assays were performed in 102 patients who underwent bilateral neck explorations or targeted parathyroidectomy for pHPT. The operation was considered complete when both an IOqPTH drop >50% and a FS diagnosis of parathyroid adenoma were obtained. RESULTS: Cure was achieved in all patients. Potential pitfalls for successful operation were encountered in 14 patients with multiglandular diseases and in 4 patients who had nonparathyroid tissue removed. FS correctly predicted the definitive histologic diagnosis with an accuracy of 81%. FS failures potentially misguided the operative therapy in 19% (14 insufficient explorations and 5 unnecessarily prolonged explorations), while IOqPTH identified all potential pitfalls and correctly guided the operative strategy, suggesting further exploration, in 100% of cases (P < .0001). After bilateral neck exploration, FS and IOqPTH correctly guided operative strategy in 86% and 100% of cases, respectively (P < .05), but both techniques were never indispensable, because potential pitfalls were already evident by macroscopic intraoperative appearance. The turnaround time and costs for IOqPTH were lower (P < .001). CONCLUSIONS: The role of FS should be reconsidered, since it can misguide the operative strategy. IOqPTH is indispensable for a focused approach and, although unnecessary in bilateral neck exploration, is more useful and cost effective than FS. PMID- 16360405 TI - Preoperative evaluation of thyroid nodules with 18FDG-PET/CT. AB - BACKGROUND: Fluorine-18-fluorodeoxyglucose positron emission tomography/computed tomography ((18)FDG-PET/CT) has become an important tool in the postoperative management of de-differentiated thyroid cancer. The utility of this imaging modality in the preoperative assessment of thyroid nodules is unclear. This study was designed to determine whether (18)FDG-PET/CT improves the preoperative diagnosis of thyroid nodules. METHODS: A total of 31 patients with 48 lesions underwent fine-needle aspiration and (18)FDG-PET/CT before surgical resection of thyroid nodules. PET/CT images were obtained 1 hour after intravenous administration of (18)FDG. Standard uptake values were calculated for regions of increased (18)FDG uptake. CT scans were evaluated to identify thyroid pathology. Final pathologic diagnoses were compared with PET/CT findings. RESULTS: Fifteen of 48 lesions were malignant and 33 were benign. Nine of 15 malignant lesions were (18)FDG-avid (sensitivity 60%). Thirty of 33 benign lesions were (18)FDG cold (specificity 91%). Positive and negative predictive values were 75% and 83%, respectively. CONCLUSIONS: (18)FDG-PET/CT provides a high negative predictive value for malignancy, making this a potentially useful tool in the evaluation of thyroid nodules with indeterminate fine-needle aspiration. However further studies with larger sample sizes are needed to determine the true efficacy of this test. PMID- 16360406 TI - Feasibility and benefit of fluorine 18-fluoro-2-deoxyglucose-guided surgery in the management of radioiodine-negative differentiated thyroid carcinoma metastases. AB - BACKGROUND: Fluorine 18-fluoro-2-deoxyglucose (FDG) positron emission tomography ((18)F-FDG PET) can be used to visualize metastases in patients with differentiated thyroid carcinoma that does not take up radioiodine ((131)I). This study was aimed at evaluating the feasibility of (18)F-FDG radio-guided surgery in patients with radioiodine-negative differentiated thyroid cancer. METHODS: Ten patients received a mean activity of 265 MBq of (18)F-FDG 30 minutes before operation. Radioactivity uptake (counts per second [cps], with a pretime of 10 seconds) in tumor and normal tissues was measured before and after resection. RESULTS: Patients with 1 to 5 foci detected by FDG-PET were included in the study. Six of these patients were injected with recombinant human thyroid stimulating hormone (TSH) preoperatively. Abnormal findings detected by preoperative (18)F-FDG PET were also detected with the gamma probe in all patients. The mean tumor activity in vivo was 3,272 cps, and tumor-to-neck and tumor-to-shoulder ratios were, respectively, 1.40 and 1.73. Tumor resection was incomplete in 3 patients. When resection was complete, mean radioactivity at the tumor site was decreased by 22%. The ex vivo mean tumor-to-normal tissue ratio was 2.4. All positive tissues detected with the probe were confirmed histologically to be differentiated thyroid cancer. The surgeon's hands were exposed to 90 to 270 microSv. CONCLUSIONS: These results show the feasibility and benefit of (18)F-FDG radio-guided surgery with a gamma probe in the management of differentiated thyroid cancer patients with radioiodine-negative recurrence. PMID- 16360407 TI - Intraoperative neurophysiology testing of the recurrent laryngeal nerve: plaudits and pitfalls. AB - BACKGROUND: Electrode-imbedded endotracheal tubes allow continuous intraoperative assessment of vocal cord function when connected to an electromyographic (EMG) response monitor. Whether this device enhances or hinders the identification and preservation of the recurrent laryngeal nerve (RLN) is unclear. METHODS: The utility of continuous intraoperative neurophysiology testing (INT) of RLNs was evaluated prospectively in 100 patients undergoing 103 thyroid or parathyroid operations, involving 185 RLNs. The initial experience with 93 RLNs was compared with the subsequent 92 RLNs. RESULTS: Overall, 97.8% of RLNs were identified intraoperatively: 1.6% visually only, 2.2% nerve stimulator only, and 94% both. There was 1 transected RLN (1.1%) in each study group. The EMG monitor could not alert the surgeon to prevent these injuries. Overall, there were 14 instances of nonfunction of visually intact RLNs (7.6%), at some point during the operation and 4 resulting in temporary paralysis (2.2%). There were 8 instances of altered RLN function (4.3%) with no altered vocal cord function postoperatively. The nerve stimulator aided dissection of the RLN in 17 instances (9.2%). There were 7 episodes (3.8%) of equipment dysfunction that hampered surgical dissection. Between study groups there was significantly increased use of the nerve stimulator to first identify the location of the RLN before visual confirmation: 4 of 93, initial group versus 25 of 92, latter group (P < .001). CONCLUSIONS: INT aids the anatomic identification of the RLN only when a positive EMG response occurs. A negative EMG response can indicate a non-nerve structure, altered function of the RLN, or equipment setup malfunction. INT cannot necessarily prevent RLN transection. PMID- 16360408 TI - Ultrasonography for the endocrine surgeon: a valuable clinical tool that enhances diagnostic and therapeutic outcomes. AB - BACKGROUND: Surgeon-performed ultrasonography (U/S) has revolutionized many subspecialties by broadening the diagnostic and interventional scope of practice. We report our experience on the impact of surgeon-performed U/S in an endocrine surgery practice. METHODS: Prospectively maintained records of patients from November 1999 to November 2004 were reviewed to establish patterns and outcomes of U/S practice. Surgeon-performed neck U/S was done routinely at the initial clinic visit and incorporated into resident/fellow education. RESULTS: A total of 5703 U/S were performed on endocrine patients with thyroid 42%, parathyroid 57%, and adrenal 1% disorders. Diagnostic fine-needle aspiration biopsy (FNA) was achieved with low sampling errors (<7%). When U/S identified thyroid nodules coexisting with hyperparathyroidism, preoperative FNA correctly established benign thyroid diagnosis and minimized need for thyroidectomy. U/S successfully imaged abnormal parathyroid glands when (99)Tc-sestamibi scans were negative. U/S data significantly changed treatment plans in nearly two thirds of thyroid cancer patients. Surgical residents readily mastered essential U/S skills. CONCLUSIONS: Surgeon-performed U/S is a highly specific tool for identification of endocrine disease in the neck. It is learned readily and performed accurately, and functions as an informative extension of physical examination. Because it substantially benefits patient care and impacts surgical decision making, neck U/S is recommended highly as a valuable adjunct to endocrine surgical practice. PMID- 16360409 TI - Medical students' perceptions of pathology and the effect of the second-year pathology course. AB - To explore the perceptions of medical students regarding pathology and the effect of the sophomore pathology course, a questionnaire was given to second-year students from 5 different medical schools at the beginning and again at the completion of their pathology course. The questionnaire was given to students in the class of 1995 and then again for those in the class of 2002, with nearly 1,500 surveys collected over the study period. The survey included questions that sought to determine students' affinity for pathology, their understanding of the typical duties of a pathologist, and how they viewed pathology relative to other specialties and perceived positives and negatives of being a pathologist. Overall, the second-year pathology course had little effect on medical students' perceptions of pathology but did provide some increase in their understanding of pathology as a profession. Responses were in general stable when comparing the different classes. Those students interested in pathology emphasized the intellectual aspects of pathology although being less deterred by negative factors such as limited patient contact. In our sample of medical schools, the sophomore pathology course was ineffective at influencing students' perceptions of pathology. Furthermore, those students who are interested in pathology are drawn because of a perceived fit between their personalities and the perception of pathology as a solely scholarly and isolated specialty. Better education about the practice of pathology in the second year, and more importantly, continuing into the clinical years, is necessary to combat these misconceptions about pathology. PMID- 16360410 TI - Focal adhesion proteins as markers of malignant transformation and prognostic indicators in breast carcinoma. AB - Focal adhesion kinase (FAK) is one of the central signaling molecules found at focal adhesion sites, which are specific areas on the cell membrane where cells attach to extracellular matrix proteins. Focal adhesion kinase interacts with multiple signaling and adaptor molecules and effects several signaling pathways. Overexpression of FAK and its substrate c-Src has been implicated in malignant transformation and acquisition of an invasive tumor phenotype of different tissues. Overexpression of the multidomain protein paxillin, which is also a FAK ligand and a c-Src substrate, has been associated with less malignant tumor behavior. The purpose of this study was to analyze the involvement of integrin signaling molecules FAK, c-Src, and paxillin in malignant transformation of the breast epithelium. Using phosphospecific antibodies FAK-pY(397) and Src-pY(416), we demonstrated that neither activation of FAK nor activation of c-Src correlates with development of invasive tumor properties. However, activation of both FAK and c-Src correlates with malignant transformation. We further demonstrated that overexpression of paxillin also correlates with malignant transformation and is a marker of a less invasive tumor phenotype. Using tissue microarray, we demonstrated that expression and activation of paxillin inversely correlated with lymph node metastases and lymphovascular invasion, respectively. No correlation between paxillin expression and activation and tumor grade, estrogen, progesterone, and Her2/Neu receptor expression was found. In summary, focal adhesion proteins FAK and c-Src can be used as markers of malignant transformation in epithelial cells but not invasive phenotype, whereas expression and activation of paxillin may represent a good prognosticator in breast carcinoma. PMID- 16360411 TI - Expression of survivin, p53, and caspase 3 in Barrett's esophagus carcinogenesis. AB - The regulation of apoptosis, as a distinctive form of programmed cell death, in multistep Barrett's esophagus (BE) carcinogenesis is poorly understood. The aim of this study was to investigate, in the intestinal metaplasia-dysplasia carcinoma sequence, the role of survivin, an inhibitor of apoptosis; the p53 protein, a tumor suppressor gene involved in cell cycle control; and caspase 3, a protease-inducing apoptosis and inhibited by survivin. Immunohistochemical expression was tested in 40 cases of BE, including 11 low-grade and 19 high-grade dysplasias (HGD), and samples were obtained from 40 surgical specimens of esophagectomy performed for HGD or Barrett's adenocarcinoma. To define the deregulation time of the proteins, overexpression was evaluated in relation to the proliferative and/or maturative compartment. In BE, cytoplasmic expression of survivin and caspase 3 (100% of cases) was significantly higher than expression of p53 (25%). The latter increased with increasing grade of dysplasia. In BE, the expression of survivin, p53, and caspase 3 mainly involved the proliferative compartment, whereas in LGD and HGD, the 3 proteins were coexpressed in both proliferative and maturative compartments. These results indicate that survivin overexpression is an early event in the proliferative compartment of BE, preceding both p53 accumulation and dysplastic changes. Cytoplasmic survivin location may indicate an initial antiapoptotic, more than proliferative, role in the early phases of Barrett carcinogenesis. Expression of caspase 3 in BE and dysplasia may be ascribed to accumulation of the nonactivated form, as the antibody used detects both cleaved and uncleaved caspase 3. PMID- 16360412 TI - Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 infection in reactive lymphoid tissues: a model for KSHV/HHV-8-related lymphomas? AB - We set out to analyze the presence of Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) in different neoplasms occurring in East Africa, a region characterized by a high KSHV/HHV-8 seroprevalence rate and endemic Kaposi's sarcoma (KS). Our results suggest that, in endemic regions of Africa, KSHV/HHV-8 is predominantly associated with KS, independently of HIV status. During the course of this study, other important information came to light. We found the presence of KSHV/HHV-8 in 2 cases of lymph nodes partially involved by Burkitt's lymphoma and KS and in 1 case of multicentric Castleman disease. Our immunophenotypic and molecular data seem to suggest 2 different mechanisms of viral infection are at work in lymphoid cells. On one hand, when B cells show a latent phase infection with KSHV/HHV-8, after the germinal center reaction, naive B cells become resting memory B cells, similarly to Epstein-Barr virus-infected B cells. On the other hand, when lytic genes such as vIL6 are expressed in naive B cells, they may be driven to differentiate into plasmablasts without undergoing germinal center reaction. Interestingly, among KSHV/HHV-8 positive cases, in those in which there was also lymphoma, the neoplastic cells were negative for KSHV/HHV-8. This further confirms that KSHV/HHV-8 is involved in the neoplastic transformation of only certain types of lymphoma, probably in relation to their precursor infected cell. In conclusion, the maturation stage of KSHV/HHV-8-positive B cells as well as the type of viral infection may well determine the morphological, phenotypic, and clinical characteristics of KSHV/HHV 8-associated lymphomas. PMID- 16360413 TI - Lesions resembling Langerhans cell histiocytosis in association with other lymphoproliferative disorders: a reactive or neoplastic phenomenon? AB - Langerhans cell histiocytosis (LCH) has been described in association with a variety of neoplasms preceding, after, or synchronous with the other tumor. In some cases, a neoplasm may arise as a complication of therapy for LCH, and in others, the association may be coincidental. Synchronous occurrence has been reported most commonly in association with malignant lymphoma in which discrete proliferations of Langerhans cells (LCs) histologically indistinguishable from LCH are seen. In most cases, these LCs are closely related to or intermingling with the primary pathology. The nature of LCs in this context remains elusive with debate as to whether they represent a true clonal neoplasm or an exaggerated reactive phenomenon. The lack of evidence for LCH progression or disease elsewhere strongly supports the latter. We have encountered 5 examples of LCH like proliferations occurring in the context of other lymphoproliferative disorders. These include 2 cases of mycosis fungoides and 1 of cutaneous B-cell pseudolymphoma, associations that to our knowledge have not been described before. Two patients were female, and the clonality of the LC proliferation was assessed using laser capture microdissection and the human androgen receptor. The results showed that the LCs forming discrete nodules in a case of cutaneous B cell pseudolymphoma and a case of Hodgkin's lymphoma were polyclonal. This suggests that, at least in a proportion of cases, the aggregates of LCs occasionally identified within other lymphoproliferative lesions represent a reactive proliferation rather than a potentially aggressive second neoplasm. PMID- 16360414 TI - A proposal for a new validated histological definition of the gastroesophageal junction. AB - Present definitions of the gastroesophageal junction (GEJ) are the point of flaring of the tubular esophagus and the proximal limit of the gastric rugal folds. Neither of these has been validated as the true GEJ. This study aims to validate the location of the true GEJ using the criterion of esophageal submucosal glands. Ten esophagogastrectomy specimens, in which there was a well defined point of flaring of the tubular esophagus that coincided with the proximal limit of gastric rugal folds, were examined by complete histological mapping to evaluate the distribution of esophageal submucosal glands and surface epithelial types. Oxyntocardiac and cardiac mucosa with or without intestinal metaplasia were present under rugal folds distal to the end of tubular esophagus in all patients to a length of 0.31 to 2.05 cm. Submucosal glands were present in the tubular esophagus and in the proximal pouch distal to the tubular esophagus in a distribution that closely coincided with squamous epithelium, oxyntocardiac, cardiac, and intestinal epithelia. Submucosal glands were never found under oxyntic mucosa. We conclude that a variable part of the saccular region distal to the tubular esophagus contains esophageal submucosal glands, therefore representing reflux-damaged distal esophagus. This results in an error, where up to 2.05 cm of distal reflux-damaged dilated esophagus can be mistaken as proximal stomach when presently accepted definitions for the GEJ are used. The true GEJ is the proximal limit of gastric oxyntic mucosa defined by histology. PMID- 16360415 TI - Involvement of adenomatous polyposis coli (APC) gene in testicular yolk sac tumor of infants. AB - The pathogenesis of testicular yolk sac tumor (YST) of infants is still unclear. Infantile YSTs rarely show isochromosome 12p or aneuploidy, which are common in adult germ cell tumors. On the other hand, recent epigenetic studies suggest the involvement of some tumor suppressor genes, including the adenomatous polyposis coli (APC) gene. In the present study, we examined 10 infantile pure YSTs for mutation, allelic loss, promoter methylation, and protein expression status of the APC gene to evaluate whether the APC gene plays a significant role in the pathogenesis of infantile YSTs. Loss of heterozygosity at 5q21, where the APC gene is localized, was detected in at least 3 (30%) of the 9 YSTs examined. None of the 10 YSTs showed mutations. Promoter methylation was detected in 7 (70%) of the 10 YSTs; among 7 YSTs showing methylation, 3 YSTs also harbored loss of heterozygosity at 5q21. Immunohistochemically, 8 infantile YSTs did not express the APC protein, whereas 2 YSTs without showing APC methylation, as well as germ cells of normal infantile testes, expressed this protein in the cytoplasm. These data indicate that inactivation of the APC gene, by allelic loss and/or promoter methylation, is related to the occurrence of infantile YSTs. PMID- 16360416 TI - Immunohistochemical and genetic analysis of Chinese nasal natural killer/T-cell lymphomas. AB - Nasal natural killer/T-cell lymphoma (N-NK/T-L) is prevalent in China. To further characterize this neoplasm, 36 cases of N-NK/T-L from 304 cases of malignant lymphomas in the north China area were investigated by histopathology, immunophenotyping, and genomic analysis of c-kit, in comparison with 11 cases of B-cell lymphoma (BCL) at the same region and 5 cases of nodal peripheral T-cell lymphoma (PTCL) (unspecified). Histopathologically, N-NK/T-L was characterized by coagulative necrosis, inflammatory background, and angiodestructive growth pattern. In 36 cases of N-NK/T-L, 27 cases (75.0%) were stained for CD45RO and 25 (72.2%) for CD3epsilon. Thirty cases (83.3%) were positive for T-cell intracellular antigen-1, 22 (61.1%) for granzyme B, 18 (50.0%) for CD56, and 11 (30.6%) for CD30, whereas none was positive for CD117. All 5 cases of PTCL displayed positive staining for CD45RO and T-cell intracellular antigen-1, 3 cases for CD3epsilon, but only 1 case for granzyme B. All 11 BCLs presented positive staining for CD20 and CD79a but negative for other antibodies. A significant relationship was observed between neoplastic cells pleomorphism and granzyme B expression (P < .05). Despite the fact that all cases were negative for CD117 staining, genomic sequences of c-kit 11 and exon 17 sequencing showed that 8 (26%) of 31 cases N-NK/T-L proved to contain genomic mutations, including 4 cases in exon 11 and 4 in exon 17. For the control group, only 1 (9%) of 11 BCLs and 1 (20%) of 5 cases of PTCL were detected to harbor mutations in exon 11. All mutations detected in 3 groups were missense by base substitution, and codes 571, 572, and 821 were hot spots. The results suggested that, in addition to histological features and routine immunophenotyping, granzyme B expression should be a more reliable marker in correct diagnosis of N-NK/T-L, and genetic analysis of c-kit mutation should be helpful in the diagnosis of this tumor. PMID- 16360417 TI - Actinomyces in infected osteoradionecrosis--underestimated? AB - Infected osteoradionecrosis (IORN) is a severe complication of radiation therapy for head and neck cancer. Infected osteoradionecrosis can lead to fracture and often requires subsequent jaw resection. It is known that irradiated bone is highly susceptible to infections, mainly with Candida species and cariogenic bacteria. Only very few data exist on Actinomyces in IORN. The study population consisted of 31 patients (7 female, 24 male; median age, 58.3 years). All patients exhibited clinical and radiological signs of IORN (infection, mucosa or skin fistula, and sequestrated bone). To detect Actinomyces colonies, histological examination was performed using several staining procedures (hematoxylin-eosin, Gram, Grocott, periodic acid-Schiff). In addition, a semi nested polymerase chain reaction (PCR) approach was designed targeting the 16S ribosomal RNA gene. We found prominent Actinomyces colonies in 20 (64.5%) of 31 patients. Most of these lesions were localized in the mandible (16/20). Most interestingly, Actinomyces were almost exclusively found attached to the necrotic bone. PCR testing confirmed the presence of Actinomyces-specific DNA sequences (Actinomyces israelii). We show that Actinomyces is considerably more frequent in IORN than previously demonstrated. We suggest that these organisms are involved in the chronic, nonhealing inflammatory processes and the purulent discharge, which are known as characteristics of IORN. It remains to be investigated whether Actinomyces could be involved in the osteolytic mechanisms. From the histopathologic perspective, detection of Actinomyces is important because these bacteria have been shown to be associated with prolonged treatment duration. PMID- 16360418 TI - CD14 and CD169 expression in human lymph nodes and spleen: specific expansion of CD14+CD169- monocyte-derived cells in diffuse large B-cell lymphomas. AB - The mononuclear phagocyte system of human lymphoid tissue comprises macrophages and dendritic cells (DCs). The heterogeneity of the non-DC mononuclear phagocyte population in human lymphoid tissue has been little addressed. Here, we studied the expression of 2 monocyte-derived markers, CD14 and CD169 (sialoadhesin), in reactive human lymphoid tissue as well as in a series of 51 B-cell lymphomas by immunohistochemistry on paraffin-embedded tissue. We confirmed that lymph node sinusoidal monocyte-derived cells were the only population staining for CD169. Although most sinusoidal histiocytes also expressed CD14, monocyte-derived cells with phagocytosis such as erythrophagocytosis, anthracosis, or tingible bodies macrophage lacked CD14 and CD169. Among B-cell lymphomas, splenic marginal zone lymphoma was the only one associated with an expansion of the CD14(+)CD169(+) cells in the cords. With respect to nodal B-cell lymphomas, CD14(+) cells were rare among B-chronic lymphocytic leukemia, follicular lymphoma (FL), mantle cell lymphoma (MCL). However, strikingly, we found a strong expansion of CD14(+)CD169( ) cells in numerous diffuse large B-cell lymphomas (DLBCLs), except in cases associated with numerous mitoses, apoptotic bodies, and tingible bodies macrophages. When cultivated in granulocyte/macrophage colony stimulating factor/interleukin 4, DLBCL purified CD14(+) cells differentiate into plasmacytoid cells, expressing DC-specific intercellular adhesion molecule 3 grabbing nonintegrin, suggesting dendritic cell differentiation potential. Our observation fits well with the lymph node and host response cluster signatures described in the gene profiling signatures of DLBCL. However, the role of this CD14(+) population that may constitute a microenvironment-related marker of this subgroup of DLBCL remains to be determined. PMID- 16360419 TI - Expression and localization of inhibitor of apoptosis proteins in normal human tissues. AB - The family of inhibitor of apoptosis (IAP) proteins can suppress apoptosis induced by a variety of triggers. Among the IAPs, cIAP1, cIAP2, and XIAP have been characterized as inhibitors of specific caspases, and their expression, together with that of survivin, has been shown in several studies to play a role as tumor marker and prognostic factor for the survival of patients with cancer. Although survivin is usually not expressed in normal adult tissues, cIAP1, cIAP2, and XIAP have been found broadly expressed at messenger RNA level within normal cells. Here, we report an immunohistochemical study in a comprehensive panel of normal human tissues, and we confirm at the protein level the wide expression of IAPs. These results are consistent with a physiological role of IAPs in normal cells. Moreover, we show that IAPs' expression levels and localization patterns differ depending on the cell lineage. The variable subcellular localization of the IAPs within different cell types suggests that compartmentalization may contribute to regulate their function. The physiological role of these proteins should be further investigated to help tailor IAP-targeted therapeutic strategies for patients with cancer and circumvent possible side effects. PMID- 16360420 TI - Histological chorioamnionitis and neonatal leukemoid reaction in low-birth-weight infants. AB - We investigated whether histological chorioamnionitis (HCA) is a risk factor predisposing to leukemoid reaction (LR) and whether LR is associated with the preterm parturition syndrome and the systemic fetal inflammation response syndrome. A prospective histological study on placentas was performed in preterm infants (5), and 86% specific for nocturnal desaturation >4% below baseline. Presence of tongue scalloping also showed PPV of 67% for abnormal AHI, 89% for apnea or hypopnea, and 89% for nocturnal desaturation. Presence and severity of tongue scalloping showed positive correlation with increasing Mallampati and modified Mallampati airway classification. CONCLUSIONS: In high-risk patients we found tongue scalloping to be predictive of sleep pathology. Tongue scalloping was also associated with pathologic polysomnography data and abnormal Mallampati grades. We feel the finding of tongue scalloping is a useful clinical indicator of sleep pathology and that its presence should prompt the physician to inquire about snoring history. PMID- 16360523 TI - Comparison of the effectiveness and safety of radiofrequency turbinoplasty and traditional surgical technique in treatment of inferior turbinate hypertrophy. AB - OBJECTIVES: Inferior turbinate hypertrophy is one of the major causes of nasal airway obstruction. Medical treatment often produces insufficient improvements. In these cases, surgical reduction of inferior turbinates can be proposed. Many different techniques are currently available. We prospectively evaluate the safety and effectiveness of radiofrequency volumetric tissue reduction (RFVTR) compared with the traditional surgical technique. METHODS: The study was conducted on 3 groups of 75 patients with symptoms and signs of nasal obstruction associated with inferior turbinate hypertrophy refractory to medical therapy. In group A, the turbinoplasty (TP) was performed using the classical surgical submucosal resection; in group B, the RFVTR was applied to inferior turbinate; and group C patients were not treated and served as control subjects. Nasal endoscopy, visual analogue scale (VAS), anterior active positional rhinomanometry, and saccharin tests were used to assess treatment outcomes at the end of week 1 and months 1 and 3 after surgery. RESULTS: Turbinate edema and secretions decreased significantly (P < 0.05) in groups A and B from 1 month after surgery. The secretions in group A increased temporarily on the seventh day after surgery. Concerning the nasal obstruction and related symptoms, significant improvement was observed at 1 month after treatment in all patients (P < 0.05) and continued up to 3 months after surgery (P < 0.0001). Rhinomanometric measurements demonstrated a significant nasal flow increase at 3 months (P < 0.0001). The nasal mucociliary transport time increased in group A at week 1. The difference among the 3 groups at month 1 was observed not significant. CONCLUSION: In this study, we demonstrated that both RFVTR and TP are effective in improving nasal obstruction and related nasal symptoms. In support of the RFVTR, different factors are important: it can be performed in local anaesthesia; it does not require a nasal package; it does not cause either a change of mucociliary function or an increase of secretions and crusts; and the patient can be discharged immediately after treatment. Therefore, we suggest that the RFVTR offers an efficient, gentle, and function-maintaining alternative to TP. However, because of the short follow-up, future investigations are needed for a more exhaustive evaluation of equivalency of the 2 turbinate procedures. PMID- 16360524 TI - Metastatic renal cell carcinoma presenting in the external auditory canal. PMID- 16360525 TI - Apocrine hydrocysadenoma of the ear. PMID- 16360526 TI - Branchio-oculo-facial syndrome with ectodermal parathyroid tissue. PMID- 16360527 TI - Histologically benign pleomorphic adenoma of parotid with subcutaneous metastases. PMID- 16360528 TI - Mucinous and squamous metaplasia in benign tumors of the parotid gland: a potential pitfall in the diagnosis. PMID- 16360529 TI - Inferior turbinate osteoma: a rare cause of nasal obstruction. PMID- 16360530 TI - ACE inhibitor associated recurrent intermittent parotid gland swelling. PMID- 16360531 TI - Deafness after bacterial meningitis: an emergency for early imaging and cochlear implant surgery. PMID- 16360532 TI - Surgical management of Meniere's disease in the era of gentamicin. PMID- 16360534 TI - Delaying postoperative radiotherapy in advanced laryngeal cancer. PMID- 16360535 TI - Anemia during pregnancy and in the postpartum: intravenous iron therapy revisited. PMID- 16360538 TI - Anemia of pregnancy treated with intravenous iron. 1951. PMID- 16360540 TI - Intravenous iron in the treatment of anemia of pregnancy. 1949. PMID- 16360542 TI - Gemcitabine-taxane experience in the treatment of metastatic breast cancer. AB - Management of metastatic breast cancer (MBC) is difficult and overall response rates (ORR) resulting from anthracycline and taxane-based regimens remain modest. The antimetabolite drug gemcitabine has been shown to have high activity when used as first-line treatment of MBC, particularly when incorporated into combined therapy regimens. Gemcitabine-containing regimens have also been used successfully as salvage therapy in women with anthracycline or taxane-pretreated MBC. Phase II clinical studies have demonstrated high ORR with gemcitabine docetaxel (ORR: 36-65.5%) and gemcitabine-paclitaxel (ORR: 40-68%) combination regimens. A highly favourable risk-benefit ratio has also been reported for gemcitabine-containing triplets such as gemcitabine-paclitaxel-epirubicin (ORR: 92%) and gemcitabine-paclitaxel-doxorubicin (ORR: 80.4%). Gemcitabine-containing regimens have a favourable toxicity profile with few serious toxic events reported. An important step forward in the evaluation of novel chemotherapeutic regimes for MBC is establishing a correlation between disease markers and response to therapy. Preliminary data suggest that there is a close relationship between HER2 extracellular domain levels (>30 ng/ml) and treatment outcome. HER2 positive patients had a lower ORR to gemcitabine-paclitaxel chemotherapy than women who were HER2-negative. Further studies to establish a link between other breast cancer markers and predicted response to treatment are warranted. PMID- 16360543 TI - Gemcitabine plus docetaxel: a new treatment option for anthracycline pretreated metastatic breast cancer patients? AB - Antimetabolite-taxane combinations have been shown to be effective chemotherapy in patients with metastatic breast cancer (MBC). A multicentre phase III trial comparing gemcitabine-docetaxel with capecitabine-docetaxel in women with anthracycline pretreated MBC was carried out in several European countries. Its results were presented recently [1]. One-hundred and fifty-three patients were randomly assigned to receive docetaxel (75 mg/m2, day 1) plus gemcitabine (1000 mg/m2, days 1, 8) and 152 received docteaxel plus capecitabine (1250 mg/m2, bid days 1-14) every 3 weeks until disease progression. No difference in efficacy of the two treatment regimens was observed in terms of progression-free survival (35 weeks in both treatment arms), overall response rate (32% vs. 32%), time to treatment failure (19 vs. 18 weeks, respectively), or response duration (36 vs. 42 weeks). Drug-related toxicity, particularly hand-foot syndrome, mucositis and diarrhoea, was more frequent with capecitabine-docetaxel and there was a higher incidence of drug-related treatment withdrawals with this combination. Gemcitabine-docetaxel appeared to have a more favourable risk-benefit profile than capecitabine-docetaxel, and is an important new treatment option for women with anthracycline-pretreated MBC. PMID- 16360544 TI - What is the best schedule for administration of gemcitabine-taxane? AB - Until recently, standard adjuvant chemotherapy for metastatic breast cancer (MBC) consisted of anthracycline-based regimens, followed by a taxane. However, data suggest that taxane-based combinations can be more effective than taxanes alone for the second part of treatment. Synergy between paclitaxel and gemcitabine was demonstrated in vitro when paclitaxel was followed by gemcitabine. Dose-dense regimens administered every 2 weeks are more effective than standard 3 weekly regimens. In a phase II study, gemcitabine plus paclitaxel every 2 weeks as first line chemotherapy of MBC was associated with an overall response rate (ORR) of 71%. Women with HER2 ECD-positive tumours have a poor ORR (40%) to first-line chemotherapy. The addition of trastuzumab to dose-dense paclitaxel-gemcitabine as first-line chemotherapy in women with HER2-positive MBC was associated with a dramatic increase in ORR to 78%, with no serious toxicity observed. Two phase III clinical trials of gemcitabine-paclitaxel as adjuvant chemotherapy in women with histologically-confirmed MBC are currently underway. Preliminary data show that this drug combination is well-tolerated, and the efficacy results are eagerly awaited. PMID- 16360545 TI - Gemcitabine in patients with ovarian cancer. AB - Standard first-line treatment of ovarian cancer (OC) consists of platinum-taxane combined chemotherapy. However, this regimen only cures about 25% of women with OC. Phase II studies have shown that platinum-gemcitabine doublet and platinum taxane-gemcitabine triplet regimens are active first-line chemotherapy in advanced OC, with overall response rates (ORR) above 55%. Several phase III studies of gemcitabine-based doublet and triplet chemotherapy in OC are currently underway. Preliminary data show that these regimens are well-tolerated, with manageable haematological toxicity, and the efficacy results are eagerly awaited. Gemcitabine is also active as second-line monotherapy in women with recurrent OC, and studies combining gemcitabine with paclitaxel, docetaxel, liposomal doxorubicin or topotecan resulted in higher ORR than gemcitabine alone. Gemcitabine-cisplatin and gemcitabine-carboplatin are active in women with platinum-resistant recurrent OC suggesting in vivo synergy between these two classes of drug. These studies show that gemcitabine-based chemotherapy may have an important role as second-line treatment in women with platinum-resistant OC. Gemcitabine combinations are also highly recommended as they avoid the problems of neurotoxicity and alopecia seen with other regimens. In order to respect the quality of life of women with recurrent OC, assessment of prognostic factors is recommended so that the most appropriate chemotherapy can be administered. PMID- 16360546 TI - How to maximize the efficacy of taxanes in breast cancer. AB - The life-expectancy of women with metastatic breast cancer (MBC) is closely linked to response to therapy. A significant increase in progression-free survival (PFS) and overall survival (OS) has been demonstrated in women who achieve a complete response. Anthracycline combinations have been proven as highly effective in MBC, and anthracycline regimens plus cyclophosphamide with or without fluorouracil were established as first-line chemotherapy for MBC in the 1990s. Clinical trials have shown that anthracycline-taxane combinations are more effective than anthracyclines or taxanes alone in terms of overall response rates (ORR), PFS and OS in women who have not received prior anthracycline chemotherapy. The use of anthracycline-based regimens is limited, however, by the widespread use of anthracycline adjuvant therapy and the development of anthracycline-resistance. Platinum-taxane combinations have similar efficacy to anthracycline-based regimens and are well-tolerated by patients. Carboplatin combined with paclitaxel or docetaxel is more effective than carboplatin or taxanes alone, with ORR of 53-62%. Taxane combinations with gemcitabine or capecitabine are also more effective than docetaxel, paclitaxel, capecitabine or gemcitabine administered alone. The efficacy of docetaxel and paclitaxel can be increased, and drug-related toxicity decreased, by adapting dose-dense schedules of drug administration. The addition of trastuzumab to taxane-based chemotherapy increases the efficacy of taxane-based regimens in women with HER2-positive MBC. PMID- 16360548 TI - An overview of the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program: 1997-2004. AB - This overview provides a summary of the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program over an 8-year period from 1997 to 2004. The evolution of the MYSTIC Program is described, as well as its design compared with other surveillance programs. In addition, the global MYSTIC Program data, published to date, are summarized, and the empiric use of carbapenems, their current indications, and meropenem usage versus resistance was discussed. From 1997 to 2004, 120 medical centers that were actively prescribing meropenem in 32 countries worldwide participated in the program. The MYSTIC Program results demonstrate the sustained potency and continued effectiveness of meropenem globally against clinically relevant Gram-negative and Gram-positive pathogens including extended spectrum beta-lactamase- and AmpC beta-lactamase-producing organisms, which may also display resistance to the fluoroquinolones and/or aminoglycosides. Furthermore, in centers actively prescribing meropenem, resistance to meropenem is not increasing despite greater resistance among the comparator antimicrobial agents. Thus, antipseudomonal carbapenems such as meropenem and imipenem remain an effective treatment option. PMID- 16360549 TI - Prevalence and antimicrobial susceptibility data for extended-spectrum beta lactamase- and AmpC-producing Enterobacteriaceae from the MYSTIC Program in Europe and the United States (1997-2004). AB - This article presents prevalence and susceptibility data from the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program in Europe (1997-2004) and the United States (1999-2004) for Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBL) and/or AmpC beta-lactamases. For ESBL producing isolates, the prevalence of Escherichia coli and Klebsiella spp. in Europe and Enterobacter spp. in the United States increased over time. For AmpC producing isolates, the prevalence of Enterobacter spp. and Citrobacter spp. decreased over time in Europe and the United States, respectively. Compared with other antimicrobial agents, meropenem and imipenem had greatest activity against ESBL-producing E. coli and Klebsiella spp. in both Europe (96.9-100.0%) and the United States (100.0%). Such activity was also found for AmpC-producing Enterobacteriaceae in Europe (50.0-100.0%), and Enterobacter spp. and Citrobacter spp. from the United States (100.0% for both). The continued efficacy of carbapenems such as meropenem confirms that these remain first-line agents for treatment of nosocomial infections caused by Enterobacteriaceae-producing ESBL or AmpC beta-lactamases. PMID- 16360550 TI - Activity of meropenem and comparators against Pseudomonas aeruginosa and Acinetobacter spp. isolated in the MYSTIC Program, 2002-2004. AB - This study examines the susceptibilities of meropenem and other broad-spectrum antimicrobials tested against bacterial isolates collected from hospitalized patients during 2002-2004 from worldwide medical centers participating in the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program. The in vitro activity of meropenem and 5 comparator antimicrobial agents was assessed against Pseudomonas aeruginosa and Acinetobacter spp. Generally, the susceptibility of Australasian and North American isolates was higher than that of the European and South American isolates. The rank order of activity of the antimicrobial agents tested against a worldwide collection of P. aeruginosa was piperacillin/tazobactam (77.7% susceptible) > meropenem (75.4%) > ceftazidime (70.0%) > imipenem (69.7%) > gentamicin (66.1%) > ciprofloxacin (62.0%). Against a worldwide collection of Acinetobacter spp. meropenem (76.1% susceptible) was the most active compound followed by imipenem (74.7%) > gentamicin (51.9%) > ciprofloxacin (40.5%) > piperacillin/tazobactam (39.8%) > ceftazidime (38.1%). The carbapenems appear to be a valuable option for the treatment of serious nosocomial infections caused by P. aeruginosa or Acinetobacter spp. over a broad geographical region. PMID- 16360551 TI - Use of a program-specific website to disseminate surveillance data obtained from the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Study. AB - The Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program is a multicenter, longitudinal surveillance study initiated in 1997 that compares the activity of meropenem and comparator antimicrobials in centers that are actively prescribing meropenem. Antimicrobial surveillance programs, such as MYSTIC, provide important information on the emergence of bacterial resistance and the nature of the underlying mechanisms in different geographical regions. Data concerning these mechanisms and patterns of antimicrobial resistance allow the implementation of changes in antimicrobial prescribing practices and infection control interventions. Therefore, it is important that data from surveillance programs be analyzed and accessible to clinicians within reasonable, clinically useful, intervals. In this article, the use of a website designed to disseminate data collected from the MYSTIC program is described. This web-based program allows specific surveillance data to be examined in a timely manner and can assist the clinician when choosing the most appropriate antibiotic for the situation encountered and hopefully maximize successful clinical outcomes. PMID- 16360552 TI - Making the most of surveillance studies: summary of the OPTAMA Program. AB - Antibiotic surveillance studies lack consideration of pharmacodynamics and provide little information about optimal dosing. By using minimum inhibitory concentration (MIC) data derived from a global surveillance study and Monte Carlo simulation, the Optimizing Pharmacodynamic Target Attainment using the MYSTIC Antibiogram (OPTAMA) Program was established to impart greater understanding of the ability to attain pharmacodynamic exposure for specific dosing regimens and their relationship with percent susceptibility. Early OPTAMA studies focused on determining the cumulative fraction of response (CFR) for various antibiotics against Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa regionally in Europe and the Americas. Later reports considered the prevalence of specific bacteria causing infections to estimate the CFR for empiric therapy of pneumonia, bloodstream, complicated skin/skin structure, and intra-abdominal infections. Collectively, the approach of the OPTAMA Program provides a novel tool that complements susceptibility data to help in the selection of appropriate empirical antibiotic therapy at the national, regional, and institutional level. PMID- 16360553 TI - Real-time pneumolysin polymerase chain reaction with melting curve analysis differentiates pneumococcus from other alpha-hemolytic streptococci. AB - The majority of pneumococcal isolates can be identified by the conventional methods based on phenotypic characteristics. Occasionally, however, the differentiation of alpha-hemolytic streptococci from pneumococci, especially those isolated from nasopharynx, is problematic due to the discrepant results obtained by the conventional identification methods. Several gene technological methods based on the amplification of genes encoding pneumococcal virulence factors, such as pneumolysin (ply) and autolysin, have been used as additional identification methods. Recent studies have shown that especially the ply gene is frequently also present in nonpneumococcal alpha-hemolytic streptococci. In this study, we compared the commonly used phenotypic identification methods with nucleic acid-based methods, commercial AccuProbetrade mark, conventional pneumolysin polymerase chain reaction (Ply-PCR), and real-time Ply-PCR in the identification of alpha-hemolytic streptococcal strains isolated from 100 consecutive nasopharyngeal specimens. We also studied if melting curve analysis and sequencing of the amplification products of a ply gene fragment could be helpful in the identification. Our results suggest that the ply gene present in alpha-hemolytic streptococci differs from that present in pneumococcus, and that melting curve analysis would prove useful in the differentiation of these bacteria. PMID- 16360554 TI - An investigation of clinical studies suggests those with multiple objectives should have at least 90% power for each endpoint. AB - BACKGROUND AND OBJECTIVES: Many clinical studies have more than one objective, either formally or informally, but this is not usually taken into account in the determination of the sample size. We investigated the overall power of a study, that is, the probability that all the objectives will be met. METHODS: We calculated the overall power in the case that the study has two primary outcome variables and in the case that one outcome variable is evaluated on two subsets, in particular, the Per Protocol group and the Intention to Treat group. RESULTS: A power of 80% for each of the two end points leads to poor power for the end points combined. However, a power of 90% preserves better the overall power. The power of the Per Protocol analysis can be higher or lower than the power of the Intention to Treat analysis. CONCLUSION: Power should be calculated for all end points combined, and it should be at least 90% for each primary end point. If the sample size for the intention-to-treat analysis is determined by adding a percentage of "nonevaluable subjects" to the sample size required for the per protocol analysis, then this may lead to an underpowered study. PMID- 16360555 TI - Imputing missing standard deviations in meta-analyses can provide accurate results. AB - BACKGROUND AND OBJECTIVES: Many reports of randomized controlled trials (RCTs) fail to provide standard deviations (SDs) of their continuous outcome measures. Some meta-analysts substitute them by those reported in other studies, either from another meta-analysis or from other studies in the same meta-analysis. But the validity of such practices has never been empirically examined. METHODS: We compared the actual standardized mean difference (SMD) of individual RCTs and the meta-analytically pooled SMD of all RCTs against those based on the above mentioned two imputation methods in two meta-analyses of antidepressants. RESULTS: Two meta-analyses included 39 RCTs of fluoxetine (n = 3,681) and 25 RCTs of amitriptyline (n = 1,832), which had actually reported means and SDs of the Hamilton Rating Scale for Depression. According to either of the two proposed imputation methods, the agreement between actual SMDs and imputed SMDs for individual RCTs was very good with ANOVA intraclass correlation coefficients between 0.61 and 0.97. The agreement between the actual pooled SMD and the imputed one was even better, with minimal differences in both their point estimates and 95% confidence intervals. CONCLUSION: For a systematic review where some of the identified trials do not report SDs, it appears safe to borrow SDs from other studies. PMID- 16360556 TI - Refill persistence with chronic medication assessed from a pharmacy database was influenced by method of calculation. AB - BACKGROUND AND OBJECTIVES: In literature, different methods of calculating persistence are used. In this study, the effect of using these different methods on persistence and the association of patients characteristics and persistence are assessed. METHODS: The PHARMO record linkage system was used to calculate persistence with antihypertensive drugs for a cohort of 14,466 new users of antihypertensives. Three different types of methods were used to define the maximum gap allowed between two prescriptions that a patient may have to be defined as a continuous user, one based on a defined number of days (varying from 9-365 days), the second based on the duration of the last prescription (varying from 0.1-4 times the duration), the third based on a combination of both methods, whichever leads to the lowest number of days. RESULTS: Refill persistence varied between 19.7-86.4% (method 1), between 27.9-90.2% (method 2), and between 19.7 86.4% (method 3). Furthermore, patient characteristics associated with persistence differed between and within the three different methods. CONCLUSION: The method used and the variation within a method influenced both persistence and the association between patient characteristics and persistence. Results of persistence studies are highly influenced by the researchers' method of the maximum allowed treatment gap. PMID- 16360557 TI - Explained variation in a model of therapeutic decision making is partitioned across patient, physician, and clinic factors. AB - BACKGROUND AND OBJECTIVE: Data on therapeutic decision making have a multilevel structure that can include patient-, provider-, and facility-level variables. A statistical method is presented for attributing explained variation in patient care to different levels of aggregation in a multilevel model with the aim of prioritizing and targeting quality improvement interventions. STUDY DESIGN AND SETTING: The proposed method is used in an analysis of adherence to evidence based guidelines for the care of patients at risk of osteoporosis. Explained variation from a multilevel model of appropriate care is partitioned across patient-, physician-, and clinic-level factors. RESULTS: The combination of patient, physician, and clinic factors explained 20.0% of the variation in patient care. Individual physician effects explained 14.0% of the variation in the data; however, more than half of this explained variation could have been attributed to the individual clinic effect. Patient fixed effects alone explained 13.4% of the variation in the observed clinical decisions. CONCLUSION: The proposed approach is an intuitive and statistically valid method for attributing explained variation in a multilevel analysis of therapeutic decision making. PMID- 16360558 TI - Models developed by three techniques did not achieve acceptable prediction of binary trauma outcomes. AB - BACKGROUND AND OBJECTIVES: To develop prediction models for outcomes following trauma that met prespecified performance criteria. To compare three methods of developing prediction models: logistic regression, classification trees, and artificial neural networks. METHODS: Models were developed using a 1996-2001 dataset from a major trauma center in Victoria, Australia. Developed models were subjected to external validation using the first year of data collection, 2001 2002, from a state-wide trauma registry for Victoria. Different authors developed models for each method. All authors were blinded to the validation dataset when developing models. RESULTS: Prediction models were developed for an intensive care unit stay following trauma (prevalence 23%) using information collected at the scene of the injury. None of the three methods gave a model that satisfied the performance criteria of sensitivity >80%, positive predictive value >50% in the validation dataset. Prediction models were also developed for death (prevalence 2.9%) using hospital-collected information. The performance criteria of sensitivity >95%, specificity >20% in the validation dataset were not satisfied by any model. CONCLUSION: No statistical method of model development was optimal. Prespecified performance criteria provide useful guides to interpreting the performance of developed models. PMID- 16360559 TI - Item response theory was used to shorten EORTC QLQ-C30 scales for use in palliative care. AB - BACKGROUND AND OBJECTIVE: The goal was to develop a shortened version of the EORTC QLQ-C30 for use in palliative care. We wanted to keep as few items as possible in each scale while still being able to compare results with studies using the original scales. We examined the possibilities of shortening the physical functioning, cognitive functioning, fatigue, and nausea and vomiting scales. STUDY DESIGN AND SETTING: The shortening was based on 2,366 (physical functioning) and 10,815 (three other scales) observations, respectively. We used item response theory to construct scoring algorithms for predicting scores on the original scales. RESULTS: Evaluations showed that a three-item physical scale, a two-item fatigue scale, and a one-item nausea or vomiting scale predicted the scores on the original scales with excellent agreement and had measurement abilities similar to the original scales with no loss or only a little loss in power to detect group differences. The results of the cognitive functioning scale indicated problems when predicting scores from a shortened version. CONCLUSION: Given the favorable results for the physical functioning, fatigue, and nausea or vomiting scales we expect that the shortened versions of these scales will be included in the abbreviated version of the EORTC QLQ-C30 for palliative care. PMID- 16360560 TI - A minimal clinically important difference was derived for the Roland-Morris Disability Questionnaire for low back pain. AB - OBJECTIVE: To compare methods commonly used to derive minimal important differences and recommend a rule for defining patients as clinically improved on the low back pain-specific Roland-Morris Disability Questionnaire (RMDQ). METHODS: 447 primary care low back pain consulters completed a questionnaire at consultation and 6 months. Patients were classified as having achieved an important change based on methods with the best theoretical qualities, that is, the standard error of measurement, reliability change index (RCI), and modified RCI (RC(indiv)), and using a 30% reduction in score from baseline. To assess clinical importance, improvements based on these methods were compared with improvements on other back pain-related measures. RESULTS: The percentage of patients rated as improved ranged from 14 to 51% by method. Using a simple rule it was possible to identify patients who had clinically important improvement (36%), patients not improved (53%), and a group of possible improvers (11%). Clinical improvement is shown if RMDQ score is reduced by 30% from baseline and back pain is rated as better on a global rating scale. CONCLUSION: A minimal clinically important difference is derived that is clinically relevant, incorporates the measurement error of the RMDQ, and allows subjects with different grades of severity to improve. PMID- 16360561 TI - Information on chemotherapy and hormone therapy from tumor registry had moderate agreement with chart reviews. AB - BACKGROUND: Surveillance, Epidemiology, and End Results (SEER) cancer registries provide accurate information on cancer surgery and radiation, but the validity of registry data on chemotherapy and hormone therapy for breast cancer has not been well studied. We validated the registry data for chemotherapy and hormone therapy against an independent medical chart review. METHODS: We identified 1,228 women diagnosed with breast cancer at age > or = 65 in 1993-1999 in the New Mexico SEER Tumor Registry and completed medical chart reviews. RESULTS: Overall, there was moderate agreement between these two databases on chemotherapy that was received within 6 months of diagnosis. The observed agreement was 96.0%, with a kappa of 0.72 (95% confidence interval: 0.64-0.79). The sensitivity of the registry data for chemotherapy was 70.7% and the specificity was 98.2%. The positive predictive value of the registry data for chemotherapy was 77.8%. The sensitivity of the registry data for hormone therapy was 59.7%, and the specificity was 89.5%. The observed agreement for hormone therapy was 80.0%, with a kappa of 0.52 (0.46 0.57). CONCLUSION: Agreement on chemotherapy and hormone therapy between the New Mexico SEER Tumor Registry and chart reviews was moderate. The preferred approach would be to combine data from different sources to obtain more complete information. PMID- 16360562 TI - The Danish National Hospital Register is a valuable study base for epidemiologic research in febrile seizures. AB - BACKGROUND AND OBJECTIVES: We examined the quality and completeness of the discharge diagnosis of febrile seizures in the Danish National Hospital Register (DNHR). METHODS: We invited all children born at the Department of Obstetrics, Aarhus University Hospital, Denmark, between January 1990 and April 1992, to participate in the study. We collected information on febrile seizures in March 2001 by mailing a questionnaire to the mothers. Reported febrile seizures were verified by telephone interviews or review of medical records. We linked the cohort to the DNHR by means of a unique civil registry number assigned to all Danish citizens at birth. Overall, 6,624 (73%) children participated in the study. RESULTS: We found that 323 (4.9%) children in the cohort had had febrile seizures, and 231 of those were registered in the DNHR (completeness: 71.5%, 95% CI: 66.3-76.4). Furthermore, we confirmed the diagnosis in 231 of 249 children registered with febrile seizures in the DNHR (predictive value of a positive registration: 92.8%, 95% CI: 88.8-95.7). CONCLUSION: The Danish National Hospital Register is a valuable tool for epidemiologic research in febrile seizures. PMID- 16360563 TI - Using cancer registry data for survival studies: the example of the Ontario Cancer Registry. AB - BACKGROUND AND OBJECTIVES: The Ontario Cancer Registry (OCR) is a population based tumor registry created to provide data for epidemiologic research and for cancer surveillance. Recently it has been used for health services research. The objective of this project was to assess the quality of the OCR data that is used in survival analysis. METHODS AND DESIGN: Clinical information for 898 patients with squamous carcinoma of the head and neck including index tumor site, date of diagnosis, vital status, date of death, and cause of death from a prospective database at the Kingston Regional Cancer Center is compared to the same data elements in the OCR for the same patients. RESULTS: There is no statistically significant difference in disease-specific survival between the information from the two databases (log rank P = .89). The OCR captured and correctly assigned index tumor site for 81.4% (detection rate). The site assignment was accurate 90.9% of the time (confirmation rate), there was agreement on vital status (dead vs. alive) for all but one patient, and there was excellent agreement on date of death. However, cause of death (cancer vs. noncancer) based on death certificates had a 31% error rate. CONCLUSION: Researchers can be confident in the survival analysis generated from data in this registry, but need to be aware of potential sources of error. PMID- 16360564 TI - In a randomized controlled trial, patients preferred electronic data collection of breast cancer risk-factor information in a mammography setting. AB - BACKGROUND AND OBJECTIVE: We evaluated patient acceptance of an electronic questionnaire to collect breast cancer risk-factor data in a mammography setting. METHODS: We developed an electronic questionnaire on a tablet computer incorporating prefilled answers and skip patterns. Using a randomized controlled study design, we tested the survey in a mammography clinic that administers a paper risk-factor questionnaire to every woman at her screening mammogram. We randomized 160 women to use the electronic survey (experimental group, n = 86) or paper survey (control group, n = 74). We evaluated patient acceptance and data completeness. RESULTS: Overall, 70.4% of the experimental group women thought the survey was very easy to use, compared to 55.6% of women in the control group. Ninety percent of experimental group women preferred using the tablet, compared to the paper questionnaire. Preference for the tablet did not differ by age; however, women > or = 60 years did not find the tablet as easy to use as did women < 60 years. The proportion of missing data was significantly lower on the tablet compared to the paper questionnaire (4.6% vs. 6.2%, P = .04). CONCLUSION: Electronic questionnaires are feasible to use in a mammography setting, can improve data quality, and are preferred by women regardless of age. PMID- 16360565 TI - The diagnosis of a smoking-related disease is a prominent trigger for smoking cessation in a retrospective cohort study. AB - OBJECTIVE: We evaluated the impact of demographic factors, smoking patterns, and the occurrence of smoking-related diseases on smoking cessation, with a particular emphasis on the temporal relationship between diagnosis of smoking related diseases and cessation. STUDY DESIGN AND SETTING: A cohort was assembled of participants of a general health screening examination aged 50-74 years. Lifetime smoking habits and medical history were obtained by a self-administered questionnaire. In a retrospective cohort study approach, predictors of cessation among ever-smokers (n = 4,575) were identified using the extended proportional hazards model. RESULTS: Male gender, late onset of smoking, and higher educational level were predictive of cessation. However, the by far strongest predictors of cessation were diagnoses of smoking-related diseases: relative cessation rates in the year of disease occurrence were 11.2 for myocardial infarction (95% confidence interval CI = 8.9-14.0), 7.2 for stroke (95% CI = 5.1 11.6), 2.5 for diabetes mellitus (95% CI = 1.6-4.0) and 4.8 for cancer (95% CI = 3.1-7.4) relative to years before diagnosis of the respective diseases. CONCLUSION: Our results underline the key role of perceived detrimental effects of smoking for cessation. When smokers personally experience the health consequences of smoking, many permanently quit. PMID- 16360566 TI - Self-reported asthma was biased in relation to disease severity while reported year of asthma onset was accurate. AB - BACKGROUND AND OBJECTIVES: The aims of the study were to assess the accuracy of self-reported asthma and notified year of asthma onset. METHODS: The study was performed on a sample of 365 subjects, 18-60 years old, with clinically diagnosed onset of asthma between 1983 and 1986. All subjects were investigated 10 years later, in 1996, with a respiratory questionnaire about the items of asthma and year of onset. The material was analyzed with logistic regression models. RESULTS: Of the 289 subjects who returned the questionnaire, asthma was reconfirmed in 251 subjects. In a logistic regression model, asthma severity was significantly associated with confirmation of asthma. The median difference between the "true" year of onset and the reported year 10 years later, the recall period was zero, with a 10th to 90th interpercentile range of -2 to 6 years. The recall period was not associated with asthma severity, bronchial hyperresponsiveness, smoking, atopy, or sex. CONCLUSION: Self-reported asthma is biased in relation to disease severity, meaning that subjects with mild disease were less prone to report their asthma. Reported year of asthma onset among adults seems to be rather accurate, with no obvious dependent misclassifications. PMID- 16360567 TI - A Colombian survey found intensive care mortality ratios were better in private vs. public hospitals. AB - BACKGROUND: Our main outcome was to identify organizational characteristics that help to evaluate the differences between the intensive care mortality ratios adjusted by APACHE II. We incorporated the variation associated with the ranking of institutions simulating its random effects under a binomial distribution. METHODS: A nationwide survey on structure, technology, and staffing resources available in Colombian intensive care units during 1997-1998 was conducted. We collected data on admissions from 20 randomly selected adult medical and surgical intensive care units. RESULTS: The mortality ratio from the 20 intensive care units ranged from 0.59 to 2.36; 80% of the intensive care units had a mortality ratio greater than 1. All four intensive care units with the lowest mortality ratio belonged to private institutions, while four of five institutions with the highest mortality belonged to the public sector. Intensive care units in private institutions also had fewer number of beds, lower median length of stay, lower occupancy rates, higher education training for specialists and nurses and fewer emergency nonelective surgical procedures. CONCLUSION: We successfully accounted for intensive care mortality baseline differences and random effects variations. There were substantial differences between intensive care units in institution type, bed availability, technology, staffing resources, and degree of training, which may have been associated with patient outcome. These results are of crucial importance to track, detect and assess future changes. PMID- 16360568 TI - Selective migration contributes to a healthy worker effect in the farming population. PMID- 16360570 TI - Administrative database: validity of recording vs. validity of diagnosis. PMID- 16360573 TI - Aneurysms of the carotid artery. AB - Carotid artery aneurysms are an uncommon but important problem. The available data suggests that, untreated, these aneurysms lead to neurologic symptoms from embolization. Pseudoaneurysms of the carotid artery result from injury or may be the long-term sequelae of a spontaneous carotid dissection. While open surgery has been the primary mode of treatment, these aneurysms are being treated more commonly with endovascular approaches. This trend is partly the result of the increasing experience with the endovascular treatment of carotid artery stenosis. The endovascular approach offers advantages in other situations where open access to the distal extent of the aneurysm is difficult or neck radiation leads to concerns about wound healing. This article outlines the etiology, new diagnostic modalities, and treatment of aneurysms of the carotid artery. PMID- 16360574 TI - Treatment of nonaortic aneurysms in the endograft era: aneurysms of the innominate and subclavian arteries. AB - Although open aneurysm repair remains the standard of care, the advent of endoluminal techniques and stent grafts has significant potential to improve treatment of aneurysms of the innominate and subclavian arteries. Regardless of the modality chosen, therapy must be individualized and treatments tailored on the basis of anatomic and etiologic considerations. Occasionally, definitive treatment of an aneurysm with endoluminal stent-graft placement is possible for focal aneurysms or pseudoaneurysms of the innominate or subclavian arteries. In other situations, endoluminal grafting may be combined with open repair to allow for the benefits of both techniques. Ultimately long-term follow-up data are necessary to determine whether the durability of the endoluminal approach warrants more widespread use. PMID- 16360575 TI - Brachial, radial, and ulnar arteries in the endovascular era: choice of intervention. AB - To say that endovascular techniques have revolutionized treatment of aortic aneurysms is an understatement. These same techniques and principles are now being applied to peripheral aneurysms. Because of the small diameter of the arteries in the arm, the relative scarcity of true aneurysms of these arteries, and the fact that these arteries are readily accessible, open surgery remains the mainstay of therapy. On the other hand, nonsurgical approaches are playing an important role in the treatment of peripheral pseudoaneurysms and aneurysmal changes associated with arterial and venous access. This article reviews the natural history, treatment, and outcomes of aneurysms of the radial, brachial, and ulnar arteries. We will also focus on interventional treatment of pseudoaneurysms. PMID- 16360576 TI - Hepatic and splenic artery aneurysms. AB - Accounting for 80% of all visceral artery aneurysms, splenic and hepatic artery lesions are rare but potentially life threatening. Although their natural history has not been well-defined, the high mortality associated with emergent repair suggests an aggressive approach is indicated. While repair is clearly mandated in patients with a symptomatic aneurysm or contained rupture, the following asymptomatic lesions also warrant intervention: (1) splenic artery aneurysms in patients with the potential to become pregnant or requiring liver transplantation, (2) hepatic aneurysms in patients with polyarteritis nodosa or fibromuscular dysplasia, (3) splenic or hepatic artery aneurysms greater than 2.0 cm in diameter, and (4) splenic or hepatic pseudoaneurysms. Although open surgical interventions have traditionally been the mainstay of therapy, endovascular techniques have increasingly been applied to this problem. Dictated predominately by the need to maintain distal end-organ perfusion, potential therapies include open surgical ligation, autogenous reconstruction, endovascular coil embolization, or percutaneous endograft placement. While offering alternative approaches, endovascular techniques have not dramatically altered the risk-to-benefit ratio in asymptomatic lesions, and both open and percutaneous approaches present viable options for elective repair. Although explored in only a limited number of patients, the application of endovascular techniques to the symptomatic patient offers the greatest potential for significant improvements in morbidity and mortality. PMID- 16360577 TI - Aneurysms of the renal artery. AB - Increasing utilization of diagnostic imaging studies has provided newer insight into our current knowledge of visceral artery aneurysms. Because many are "silent," an appreciation of the true incidence and natural history of these nonaortic arterial aneurysms is only recently being understood. Historical data suggest that renal artery aneurysms (RAAs) occur in approximately 0.1% of the general population. Recognized complications associated with RAAs include renovascular hypertension, renal artery thrombosis, infarction from distal embolization, arteriovenous fistula formation, and, the most dreaded risk, rupture. Unfortunately, there remains continued controversy about the indications for and mode of treating RAAs. This article attempts to shed some light on the contemporary management options in view of advances in percutaneous endoluminal interventions. PMID- 16360578 TI - Isolated iliac artery aneurysms. AB - Isolated aneurysms of the iliac arteries are extremely rare, comprising less than 2% of all aneurysmal disease. These aneurysms are typically seen in older men. Their natural history, although fairly indolent, carries a significant risk of rupture when the aneurysms have attained a large size. Their operative mortality is significantly higher when undertaken as an emergent versus elective procedure, underscoring the importance of early diagnosis and appropriate management. This article reviews the literature with regard to the natural history, diagnostic workup, and treatment of iliac artery aneurysms. For patients undergoing elective repair, preoperative imaging with computed tomography or magnetic resonance is advocated. Repair is recommended for good-risk patients with aneurysms larger than 3.5 cm. A working classification based on aneurysmal anatomy is provided along with an outline of the suggested open and endovascular surgical options. Results of open and endovascular strategies are summarized and follow-up recommendations are proposed. PMID- 16360579 TI - True and false aneurysms of the femoral artery. AB - True aneurysms of the femoral artery are uncommon. They are most often identified in elderly males and are frequently associated with aneurysms at other locations. Femoral artery aneurysms that are symptomatic or larger than 2.5 cm should be repaired in order to prevent limb-threatening complications, such as rupture, thrombosis, or embolization. Open repair is the standard method of treatment and should be preceded by evaluation for coexisting aortoiliac or popliteal aneurysms, assessment of superficial femoral artery patency, and determination of the point of origin of the deep femoral artery relative to the aneurysm sac. Femoral artery pseudoaneurysms are most often seen in the setting of previous femoral artery catheterization but may also be associated with trauma, anastomotic leakage, or infection. The majority of femoral pseudoaneurysms less than 3 cm in diameter will spontaneously thrombose and may be observed with serial duplex ultrasound exams in asymptomatic patients. Symptomatic pseudoaneurysms, pseudoaneurysms with a diameter greater than 3 cm, and those found in patients who are anticoagulated should usually be treated. Ultrasound guided thrombin injection is our preferred method for treating femoral pseudoaneurysm because of its low risk, high success rate, and efficacy in the setting of anticoagulation. Ultrasound-guided compression is an alternative method that may be considered when thrombin products are contraindicated. Open pseudoaneurysm repair should be undertaken in the setting of infection, rapid expansion, or if less-invasive methods are not technically feasible. PMID- 16360580 TI - Popliteal artery aneurysms: tried, true, and new approaches to therapy. AB - Popliteal artery aneurysms are the most common peripheral aneurysm. They are associated with concomitant contralateral popliteal aneurysms and abdominal aortic aneurysms. Patients with unrecognized aneurysms may present with acute limb ischemia and potential for limb loss. Use of preoperative lytic therapy to improve distal runoff prior to exclusion and bypass has significantly improved the outcome from acute thrombosis. Long-term follow-up has shown that both proximal and distal ligation of the aneurysm are important because a thrombosed aneurysm may begin to enlarge after a long period of quiescence. Popliteal aneurysms are now being treated with covered stents. This article delineates the current evaluation, preoperative planning, and surgical and endovascular approaches to this disease. With proper diagnosis and repair, excellent limb salvage and postoperative function has become the norm. PMID- 16360581 TI - A new quantitative analysis of significant timing differences between externally cued and self-initiated motor tasks in an fMRI study. AB - It is generally accepted that the temporal resolution of blood oxygenation level dependent functional MRI is limited due to the inherent latency and longevity of the haemodynamic response. However, in this study we introduce a technique for measurement of timing differences from within the same brain region in two (or more) separate tasks that allows accurate determination of cortical timing differences 200 ms. Our technique, based on a novel use of linear regression analysis, is shown to yield accurate results both in simulated and experimental data. We show that cortical timing differences measured using fMRI are consistent with published electrophysiological results. Measurement of timing differences using this technique could prove a useful strategy for identifying neural network components in a wide range of cognitive paradigms. PMID- 16360582 TI - Diffusion tensor imaging to study anisotropy in a particular porous system: the trabecular bone network. AB - During the last decade, considerable effort has been invested into the development of diffusion tensor imaging (DTI) mainly used to investigate cerebral morphology. The aim of this paper is to review and to discuss our recent results about high magnetic field DTI application to study spongy bone tissue. Due to its peculiar properties, spongy bone represents a particular porous system sample. Strategies to perform DTI on porous systems and issues linked to DTI outcome interpretation are presented on the basis of our results concerning trabecular bone network characterization. PMID- 16360584 TI - Transcranial Doppler in acute stroke. AB - Transcranial Doppler (TCD) is an evolving neurovascular ultrasound technique that has an established diagnostic and potential therapeutic role in acute stroke management. Angiographically validated criteria for circle-of-Willis occlusion and thrombolysis in brain ischemia classification of residual flow have set the stage for the further development of this technique. TCD has shown its clinical value in thrombolysis monitoring and early emboli detection. The therapeutic effect requires confirmation and may be enhanced further by nanobubble technologies. PMID- 16360585 TI - CT perfusion in acute stroke. AB - As new treatments are developed for stroke, the potential clinical applications of CT perfusion (CTP) imaging in the diagnosis, triage, and therapeutic monitoring of these diseases are certain to increase. Technical advances in scanner hardware and software should no doubt continue to increase the speed, coverage, and resolution of CTP imaging. CTP offers the promise of efficient use of imaging resources and, potentially, of decreased morbidity. Most importantly, current CT technology already permits the incorporation of CTP as part of an all in-one acute stroke examination to answer the four fundamental questions of stroke triage quickly and accurately, further increasing the contribution of imaging to the diagnosis and treatment of acute stroke. PMID- 16360586 TI - Diffusion-weighted imaging in acute stroke. AB - In magnetic resonance diffusion-weighted imaging (DWI), regions of the brain are depicted not only on the basis of physical properties, such as T2 relaxation and spin density, which influence image contrast in conventional MR imaging, but also by local characteristics of water molecule diffusion. The diffusion of water molecules is altered in a variety of disease processes, including ischemic stroke. The changes that occur in acute infarction enable DWI to detect very early ischemia. Also, because predictable progression of diffusion findings occurs during the evolution of ischemia, DWI enables more precise estimation of the time of stroke onset than does conventional imaging. PMID- 16360587 TI - Xenon CT cerebral blood flow in acute stroke. AB - Acute stroke therapy is evolving rapidly as research moves toward extending the time window for treatment so that more patients can benefit. As physiology-based imaging increasingly is used in patient selection, it is becoming evident that rigid time windows are not applicable to individual patients. Xenon CT has an important role in acute stroke therapeutic intervention as a quantitative, reproducible, rapid, and safe modality, which can provide valuable physiologic data that can optimize patient triage and aid in management. PMID- 16360588 TI - Single-photon emission CT imaging in acute stroke. AB - Single-photon emission CT (SPECT) is an underused noninvasive imaging tool for the management of patients who have acute or chronic ischemia. SPECT was introduced in the late 1970s and is a proven, cost-effective means for the evaluation of regional cerebral blood flow and cerebrovascular reserve. Evaluation of cerebral blood flow using SPECT has become more accessible with the commercial availability of tracers that cross the blood-brain barrier and are retained by cells of the central nervous system. PMID- 16360589 TI - Collaterals in acute stroke: beyond the clot. AB - Collateral circulation is a fundamental determinant of stroke pathophysiology. Distal arterial embolism and hypoperfusion resulting from severe proximal arterial stenosis may be offset by collateral flow. Collaterals influence whether or not infarction results. The detection and characterization of arterial deoxygenation and other consequences of collateral perfusion depend on neuroimaging techniques. Imaging advances will further the understanding of clinical correlates, including collateral sustenance and collateral failure, and possibly promote the development of collateral therapeutics. Refinements of perfusion imaging protocols may quantify the delay and dispersion of collateral flow more accurately. This review explores the role of collateral flow in acute ischemic stroke and describes the imaging modalities used to investigate phenomena "beyond the clot." PMID- 16360590 TI - Extending the time window for thrombolysis: evidence from acute stroke trials. AB - Data from intravenous tissue plasminogen activator studies have shown rapidly diminishing clinical benefit beyond 3 hours when noncontrast CT is used for treatment triage. Newer trials, such as the Desmoteplase in Acute Ischemic Stroke trial, have now successfully pushed the time window out to 9 hours using the concept of penumbral imaging and treatment of the perfusion-diffusion mismatch. Advanced imaging with CT or MR imaging protocols is providing a means for rational physiologic selection and outcomes assessment in stroke treatment. PMID- 16360591 TI - Pediatric stroke: the child is not merely a small adult. AB - Arterial ischemic stroke and sinovenous thrombosis are a significant yet under recognized causes of mortality and morbidity in the pediatric population. With increasingly complex etiologies yet urgency for rapid diagnosis and treatment, pediatric stroke teams likely will become the standard of care. A common terminology must be developed to avoid confusing types of acute cerebral insults- such as focal arterial ischemic stroke and global hypoxia and ischemia--that have different causes and pathophysiologic mechanisms of injury. Increased awareness of unique pediatric stroke subtypes, their clinical presentation, and their imaging findings will facilitate early identification and development of optimal treatment strategies. PMID- 16360592 TI - Vulnerable plaque imaging. AB - The concept of vulnerable plaque is well established with increasing evidence from clinical and basic research. The paradigm has shifted from focusing exclusively on the hemodynamic effects of plaque (ie, resulting lumenal stenosis alone as a predictor of stroke risk) to assessment of the structure and composition of plaque (eg, denuded endothelium with inflammatory elements as a nidus for platelet-fibrin clumping). It is increasingly evident that methods to detect and characterize vulnerable plaque must be developed and optimized. Although MR imaging, CT, and ultrasound provide data regarding single lesions, future investigations relying heavily on nuclear medicine techniques may offer functional assessment of the entire cardiovascular system. PMID- 16360593 TI - Technical aspects of perfusion-weighted imaging. AB - There is increasing interest in using diffusion-weighted (DWI) MR imaging and perfusion-weighted MR imaging (PWI) to assist clinical decision-making in the management of acute stroke patients. Larger PWI than DWI lesions have been speculated to represent potentially salvageable tissue that is at risk of infarction unless nutritive flow is restored and presence of these mismatches have been proposed as inclusion criteria for identifying patients most likely to benefit from therapeutic intervention. Understanding the technical aspects of PWI may improve comprehension of the capabilities and limitations of this technique. PMID- 16360594 TI - Sodium MR imaging of acute and subacute stroke for assessment of tissue viability. AB - Sodium MR imaging at 3.0 T provides high-quality images in acceptable acquisition times that allow assessment of tissue viability as defined by maintenance of sodium ion homeostasis. This application is made feasible for clinical stroke evaluation by an efficient projection pulse sequence with extremely short echo time values. This twisted projection imaging provides high signal-to-noise images at adequate resolution (5 x 5 x 5 mm(3)) in less than 10 minutes at 3.0 T. The images are quantified as tissue sodium concentration (TSC) maps that can be interpreted directly in terms of tissue viability. With infarction, baseline TSC values of less than 45 mmol/L increase at variable rates to approximately 70 mmol/L, allowing monitoring of the progression of stroke pathophysiology. PMID- 16360595 TI - Diffusion tensor imaging and fiber tractography in acute stroke. AB - Diffusion tensor imaging (DTI) permits the quantitative evaluation of white matter pathology using measures of diffusion anisotropy. Fiber tractography based on DTI can reveal the three-dimensional white matter connectivity of the human brain. DTI fiber tractography is used to localize stroke lesions in relation to functionally important pathways and to assess wallerian degeneration, which may allow more accurate prognosis of long-term recovery or disability. DTI also improves the evaluation of hypoxic-ischemic injury to the developing brain of newborns and infants. DTI and fiber tractography may prove useful in elucidating alterations in brain connectivity resulting from neuroplasticity after stroke. PMID- 16360596 TI - Functional cerebrovascular imaging in brain ischemia: permeability, reactivity, and functional MR imaging. AB - MR and CT have evolved and are both capable of providing important physiologic information for guiding acute stroke therapy. This information could help to alleviate the growing unease concerning the low numbers of patients who meet the nonindividualized epidemiologically-derived enrollment criteria for thrombolysis. Physiologic assessment, such as measurement of blood-brain barrier permeability, could lead to individualization of eligibility for thrombolysis with expanded patient enrollment. Cerebrovascular reactivity analysis ("brain stress test") could improve the management of patients who have carotid occlusive disease. Detection of impaired autoregulation would favor surgical or endovascular intervention as opposed to medical management in these patients. The application of physiologic imaging techniques will undoubtedly play a key role in the management of patients who have disorders of cerebral blood flow. PMID- 16360597 TI - Stroke recovery and its imaging. AB - Functional imaging of stroke recovery is a unique source of information that might be useful in the development of restorative treatments. Several features of brain function change spontaneously after stroke. Current studies define many of the most common events. Key challenges for the future are to develop standardized approaches to help address certain questions, determine the psychometric qualities of these measures, and define the clinical usefulness of these methods. PMID- 16360598 TI - Advances in stroke neuroprotection: hyperoxia and beyond. AB - Refinements in patient selection, improved methods of drug delivery, use of more clinically relevant animal stroke models, and the use of combination therapies that target the entire neurovascular unit make stroke neuroprotection an achievable goal. This article provides an overview of the major mechanisms of neuronal injury and the status of neuroprotective drug trials and reviews emerging strategies for treatment of acute ischemic stroke. Advances in the fields of stem cell transplantation, stroke recovery, molecular neuroimaging, genomics, and proteomics will provide new therapeutic avenues in the near future. These and other developments over the past decade raise expectations that successful stroke neuroprotection is imminent. PMID- 16360599 TI - A prospective randomized controlled trial to determine if cryotherapy can reduce the pain of patients with minor form of recurrent aphthous stomatitis. AB - BACKGROUND: Tissue cooling has long been used in the management of both acute and chronic pain. OBJECTIVE: To determine whether the application of cryotherapy can reduce the pain of patients with minor form of recurrent aphthous stomatitis. STUDY DESIGN: Twenty adult patients who had 2 discrete aphthous stomatitides in the labial mucosa at the same time were included in this prospective, randomized, and placebo-controlled study. One of the 2 aphthous stomatitides was treated with cryotherapy, the other serving as a control. The pain of aphthous stomatitis was scored by the patient on a 6-point scale (from 0 to 5). The size of the aphthous stomatitis was also measured. RESULTS: At any interval, no statistical difference was found between the cryotherapy-treated aphthous stomatitis and the control in the change in the value of pain severity, nor was any statistical difference found in the change in the size of the aphthous stomatitis. However, a trend toward less pain in the aphthous stomatitis receiving cryotherapy was noted. CONCLUSION: These results suggest that application of cryotherapy on minor form of recurrent oral aphthous stomatitis has no beneficial analgesic effect compared to placebo. PMID- 16360600 TI - Endodontic treatment of dens invaginatus: a 5-year follow-up. AB - Dens invaginatus is an anomaly of the tooth formation of embryonic origin that presents itself in several morphologic types. The complex anatomy of these teeth makes nonsurgical endodontic treatment complex and more so when its apex is immature. The 2 cases reported illustrate the nonsurgical endodontic management of a dens invaginatus type II and type III with an immature apex and periapical lesions, in which mineral trioxide aggregate (MTA) in one case, and calcium hydroxide in the other one, were the materials used. A 5-year follow-up of both cases shows a complete periapical healing with bone formation at the site of the lesions. PMID- 16360601 TI - Mastery through surgical repetition. PMID- 16360602 TI - A review of odontogenic keratocysts and the behavior of recurrences. AB - OBJECTIVE: The purpose of this study is to report experiences of odontogenic keratocysts (OKCs) and analyze information regarding recurrences to better understand the nature of recurrences. STUDY DESIGN: Fifty-one cases of OKC treated at the Faculty of Dentistry, Chulalongkorn University, from 1988 to 2003 were studied retrospectively. Clinicoradiographic features, histologic features, and methods of treatment were reviewed. Recurrences were analyzed and compared with respect to sites of involvement, relationship to the remaining teeth, and methods of treatment. RESULTS: OKCs occurred predominantly in the 11- to 40-year old age group. The body-angle-ramus area of mandible was the most common site of occurrence. Radiographically, the unilocular to multilocular radiolucency ratio was 2.5:1. Multilocular lesions occurred more frequently in the mandible (P < .05). Most of the lesions were diagnosed histologically as parakeratinized OKC (93.7%). The patients were followed from 1-14.6 years. However, 20 patients were lost to follow-up after a short period of time, and recurrences were found in 7 out of 31 cases (22.6%). The recurrent tumors occurred more frequently in patients who had an OKC associated with the remaining teeth and were treated by enucleation or enucleation with adjuvant therapy. CONCLUSIONS: According to a high recurrent rate of OKCs treated by enucleation, clinicians should give more attention to the dentate area if the enucleation is chosen as the treatment of choice. PMID- 16360604 TI - Osteogenic alveolar distraction: a review of the literature. AB - OBJECTIVE: Alveolar distraction is a relatively novel procedure by which alveolar bone and underlying mucosa are regenerated. The low predictability of other vertical or horizontal bone regeneration methods has increased interest in this promising technique. This article was designed to review published clinical and experimental results on alveolar distraction, including basic research in other disciplines (maxillofacial and orthopedic distraction) related to or with influence on alveolar distraction. STUDY DESIGN: A review of the international literature was performed to summarize results of clinical and experimental studies on alveolar distraction and on distraction at other anatomical sites that contribute important findings on tissue biology, molecular mechanisms, and other factors that influence and participate in the alveolar distraction process. RESULTS: Research into alveolar distraction has addressed the latency phase, distraction phase, and consolidation phase, yielding highly variable results. Little experimental research has been carried out on this procedure, and most publications are clinical studies with a short follow-up period. Published studies have reported a high rate of complications, attributable to our current lack of understanding of the process. CONCLUSIONS: Definitive conclusions on alveolar distraction are hampered by the lack of clinical and experimental studies to date. Greater knowledge of the factors that influence the distraction process will lead to a more predictable and efficacious distraction technique and a better distractor design. PMID- 16360605 TI - The oral implications of caustic soda ingestion in children. AB - The morbidity related to caustic soda (sodium hydroxide) ingestion is well described in the literature. The majority of publications have concentrated on the effects to the trachea and gastrointestinal tract, with little reference to the oral and peri-oral areas. Accidental ingestion of sodium hydroxide-containing substances is fortunately rare; however the consequences can be devastating. Three cases of children who drank caustic substances are described. Treatment included fitting splints, injecting steroids, local surgical procedures, and the use of dynamic appliances to maintain mouth opening. Despite these interventions, all patients developed severe scarring, resulting in stenosis of the oral musculature and extra-articular ankylosis. There is perhaps a role for further investigation of early use of antiproliferative agents to prevent scarring, more aggressive surgery, and long-term physiotherapy appliance use. These patients require lifelong follow-up. PMID- 16360606 TI - Reconstruction of the severely atrophic mandible using calvarial split bone grafts for implant-supported oral rehabilitation. AB - OBJECTIVES: This article describes reconstruction of the severely atrophic mandible using calvarial bone grafts for implant-supported prosthetic oral rehabilitation. The study aim was to evaluate the efficacy of the treatment by determining implant survival and complication rates, and the extent of the postoperative graft resorption. STUDY DESIGN: Ten patients who underwent the treatment were followed clinically and radiologically using panoramic radiographs and CT scans during a mean postoperative period of 30 months. RESULTS: Good bone healing was observable 6 months postoperatively. The height reduction measured on panoramic radiographs was insignificant (mean 0.68 mm). Only minor complications occurred. Implant survival was 95%. Prosthodontic treatment was successfully performed in all cases, resulting in an improvement of oral function. Histological analysis of 1 bone biopsy showed minimal resorptive changes in otherwise very dense bone. CONCLUSION: Augmentation using calvarial grafts is a promising treatment alternative for the severely atrophic mandible. PMID- 16360607 TI - Drooling of saliva: a review of the etiology and management options. AB - Drooling of saliva appears to be the consequence of a dysfunction in the coordination of the swallowing mechanism, resulting in excess pooling of saliva in the anterior portion of the oral cavity and the unintentional loss of saliva from the mouth. Drooling can produce significant negative effects on physical health and quality of life, especially in patients with chronic neurological disabilities. Various approaches to manage this condition have been described in the literature, including oral motor therapy, behavior modification via biofeedback, orofacial regulation therapy, drug therapy, radiotherapy, and surgical treatments. Minimally invasive modalities, such as injection of botulinum toxin, photocoagulation, and acupuncture, have also been reported. This article provides a comprehensive and thorough overview of drooling, with an emphasis on understanding its etiologies and modalities of treatment. PMID- 16360608 TI - Congenital insensitivity to pain--review and report of a case with dental implications. AB - Pain is a protective mechanism for the body. Absence of pain is a symptom in several disorders, both congenital and acquired. The congenital types are present at birth and affect the number and distribution of types of nerve fibers. At present, 5 types of hereditary sensory and autonomic neuropathies have been identified. The various disorders within this group are classified according to the different patterns of sensory and autonomic dysfunction and peripheral neuropathy and the presence of additional clinical features such as learning disability. However, the field is currently moving away from classification based on clinical presentation toward classification based on underlying genetic abnormality. In the absence of pain, patients are at risk of late presentation with illnesses or injuries, and have an increased incidence of traumatic injury. Self-mutilation is an almost invariable feature of these disorders. We report the case of a patient with congenital insensitivity to pain that presented with self mutilation injuries to his hands and oral tissues caused by biting. The severe nature of these injuries necessitated serial extraction of his primary teeth soon after eruption, which led to a cessation of the problem. The mutilation has not returned following the eruption of the first of his permanent teeth, suggesting that he has learned not to bite himself, even though to do so causes him no discomfort. PMID- 16360609 TI - Association between herpes simplex virus type 1 and Helicobacter pylori in US adolescents. AB - OBJECTIVE: To understand how Helicobacter pylori infection is acquired and the role that herpes simplex virus type 1 (HSV-1) may have, we determined whether an association between HSV and H pylori exists at the individual level and for what reason. STUDY DESIGN: Data were collected from 1,090 participants aged 12-19 years during phase 1 (1988-1991) of the NHANES III. Generalized estimating equations were used to estimate prevalence ratios (PR). RESULTS: The crude overall PR and 95% CI for H pylori seropositivity comparing HSV+ to HSV- individuals was 2.20 (1.69-2.85). In large urban households the PR adjusted for poverty level and race/ethnicity was twice that in small nonurban households (2.27 versus 1.15, respectively). CONCLUSIONS: Overall, HSV-1 seropositivity is associated with a higher H pylori seroprevalence. The negligible association found in some strata suggests that shared environmental factors or routes of transmission rather than biologic reasons may be primarily responsible for this association. PMID- 16360610 TI - Mandibular osteomyelitis and tooth exfoliation following zoster-CMV co-infection. AB - Herpes zoster is a common viral infection, the oral soft tissue manifestations of which are widely known and recognized. Reports of spontaneous tooth exfoliation and jaw osteonecrosis following herpes zoster infection in the distribution of the trigeminal nerve are extremely infrequent and sporadic, with only 39 cases being reported in the literature. We report an additional case of mandibular osteomyelitis and spontaneous tooth exfoliation following herpes zoster infection, which occurred in the left mandible of a 70-year-old diabetic man; however, our case also showed CMV co-infection. The role of CMV in the pathogenesis of the osteonecrosis remains uncertain. Awareness of the possibility of CMV co-infection in various oral diseases including oral ulcers, Kaposi's sarcoma, and herpes zoster infections especially in immunocompromised patients is important, since spread of the CMV can easily occur to other sites with potentially fatal consequences. Early diagnosis can lead to effective treatment and prevention of complications. PMID- 16360611 TI - Osteogenic sarcoma of the jaw bones: a single institution experience over a 21 year period. AB - OBJECTIVE: The purpose of this article is to review cases of osteogenic sarcoma of the jaw bones diagnosed in our institution during a 21-year period. STUDY DESIGN: Records of patients seen at the Lagos University Teaching Hospital between January 1983 and December 2003 with histologic diagnosis of osteogenic sarcoma were analyzed for age and gender of patients, as well as sites and histologic types of tumor. RESULTS: Osteogenic sarcoma of the jaws constituted 0.6% of all the biopsies of oral and jaw lesions seen within the period under study. The mean (+/-SD) age of patients at presentation was 27.2 +/- 13.6 years (range 11-70 years). The mean age of patients with maxillary lesions, 30.7 +/- 15.8 years (range 17-70 years), was higher than that of patients with mandibular lesions, 23.4 +/- 10.1 years (range 11-40 years); however, no statistical significance difference was found (P = .08) There were 12 male and 5 female patients, a ratio of 2.4:1. Nine (53%) tumors were found in the maxilla and 8 (47%) in the mandible, a ratio of 1.1:1. The mandibular lesions were predominant in female patients, while the maxillary lesions were predominant in male patients. The most common histologic subtype was chondroblastic (47%), followed by fibroblastic (35.3%) and osteoblastic types (17.7%). CONCLUSION: Osteogenic sarcoma of the jaws is rare in patients who presented with tumor and tumor-like lesions of the jaws in our center, in agreement with previous reports from Africa and other centers around the world. However, the mean age at presentation of our patients was lower than reported in other parts of the world. PMID- 16360612 TI - Desmoplastic fibroma of the jaw: a case report and review of literature. AB - Desmoplastic fibroma is a benign intraosseous neoplasm that is recognized as the intraosseous counterpart of soft tissue fibromatosis in both gnathic and extragnathic sites. It has a propensity for locally aggressive behavior and local recurrence. In the present report, we define the clinicopathological and radiographic features of a desmoplastic fibroma of the mandible in an 8-year-old white boy who initially presented with a 2-month history of a rapidly expanding, painless mass along the right inferior border of his mandible. A critical and comprehensive review of the English language literature is also provided. PMID- 16360613 TI - The significance of carotid artery atheromas on panoramic radiographs in the diagnosis of occult metabolic syndrome. AB - OBJECTIVE: Metabolic syndrome (MetS), the co-occurrence of abdominal obesity, hypertriglyceridemia, reduced HDL cholesterol, hypertension, and insulin resistance promotes carotid atherosclerosis and stroke. The objective of this study was to determine if the presence of calcified atheromas detected on panoramic radiographs of individuals free of overt vascular disease may herald occult MetS. STUDY DESIGN: Ninety-four individuals (mean age 65.6 years) with a calcified atheroma detected by a VA dental clinic were evaluated. A like-aged group was used for comparative analysis. RESULTS: Fifteen percent of individuals (mean age 64 years) with an atheroma had occult MetS. Mean waist circumference was 116 cm, BMI 32.7 kg/m2, triglycerides 250 mg/dL, HDL cholesterol 35 mg/dL, blood pressure 147/87 mm Hg, and glucose 117 mg/dL. Only 6% of controls had occult MetS but this difference in prevalence was not proven to be statistically significant (P = .059). CONCLUSION: Some individuals with a calcified atheroma may have undiagnosed MetS and should be referred to their physician because aggressive management may preclude a stroke. PMID- 16360614 TI - Clinical study of preoperative angiography and embolization of hypervascular neoplasms in the oral and maxillofacial region. AB - OBJECTIVE: The purpose of this study was to investigate the angiographic features of hypervascular head and neck neoplasms and to evaluate the effects of embolization on these lesions. METHODS: Angiograms and operation records of 25 patients with hypervascular neoplasms (23 neck paragangliomas, 1 hemangiopericytoma, and 1 hemangioendothelioma) were retrospectively analyzed, and the effects of 8 embolization procedures were estimated. RESULTS: Angiograms demonstrated that 23 neck paragangliomas (NPs) were manifested as richly vascularized lesions, and were divided into 3 types. Type I NPs (n = 2) were located cranial-laterally to the carotid bifurcation, and were removed with an intact carotid artery and injured vagus nerve. Type II (n = 17) lesions widened the bifurcations in 16 of 17 cases. During the operation, the continuity of the internal carotid arteries was preserved in 15 of 17 cases. Type III (n = 4) lesions enveloped the carotid bifurcations, and were removed together with the bifurcations in 3 of 4 cases. In 6 type II paraganglioma cases intra-arterial embolization was employed and the paragangliomas were removed with less blood loss (238 mL) than the nonembolized group (600 mL). Additionally, the infratemporal hemangiopericytoma and the parotid hemangioendothelioma were embolized and removed uneventfully. CONCLUSIONS: Angiographic studies are highly valuable for the diagnosis and preoperative analysis of hypervascular head and neck neoplasms. Also, embolization therapy may minimize intraoperative blood loss. Both methods should be employed in a team approach to lesion treatment. PMID- 16360615 TI - Carotid artery calcification detected on panoramic radiographs in a group of Thai population. AB - OBJECTIVE: To determine the prevalence of carotid artery calcification (CAC) detected on panoramic radiographs in a Thai population. STUDY DESIGN: The panoramic radiographs of the patients 50 and older (N = 1,370) visiting Mahidol University from January 1998 through September 2004 were retrospectively reviewed for CAC. The medical records of the positive subjects were then reviewed. RESULTS: Thirty-four (2.5%) of the 1,370 patients, 16 men and 18 women, with a mean age of 69 and a range of 50 to 87 years, had 1 or more CACs. These calcifications were unilateral in 25 (73.5%) and bilateral in 9 (26.5%) subjects. Of those positive subjects, 18 reported hypertension, 10 reported diabetes mellitus, and 5 reported hyperlipidemia. CONCLUSIONS: Although it is uncommon to find CAC in the Thai population, dentists should be aware of this calcification on the routine panoramic radiographs and promptly refer for cerebrovascular and cardiovascular evaluation. PMID- 16360616 TI - Observations on healing following endodontic surgery in nonhuman primates (Macaca fascicularis): effects of rhBMP-2. AB - OBJECTIVES: The potential of recombinant human bone morphogenetic protein-2 (rhBMP-2) to enhance bone healing following endodontic surgery was tested. The pattern and timing of de novo bone formation and cementum regeneration, and the potential for root resorption and ankylosis to accompany bone formation were evaluated. STUDY DESIGN: Pulpal infections were induced in maxillary and mandibular incisor teeth in young adult Cynomolgus monkeys. The teeth received conventional endodontic treatment immediately followed by surgical root resection. In a randomized split-mouth design, contralateral apical bone defects received rhBMP-2 in absorbable collagen sponge (ACS) carrier or served as sham surgery controls to provide histological and radiographic evaluations following 1 (mandibular incisors) and 4.5 (maxillary incisors) month(s) postsurgery. RESULTS: At 1 month postsurgery trabecular bone filled the apical bone defects. The newly formed bone appeared considerably more mature and had assumed characteristics of the contiguous resident bone at 4.5 months postsurgery. The resected root tips were almost completely covered by new cementum with a maturing functionally oriented periodontal ligament. Localized inflammatory infiltrates were associated with the filled root canals and extruded root-filling material. Root resorption and ankylosis were not observed. There were no apparent differences in healing patterns between sites implanted with rhBMP-2/ACS and those serving as sham surgery controls. CONCLUSIONS: Under conditions where the influence of infectious elements and irritation caused by root filling material are minimized, bone formation and cementum regeneration appears rapid following endodontic surgery. rhBMP-2/ACS did not offer an obvious benefit above and beyond that of the native osteogenic potential in this animal model. PMID- 16360617 TI - Influence of instrument taper on the resistance to fracture of endodontically treated roots. AB - OBJECTIVE: The aim was to compare the force required to fracture uninstrumented and instrumented canines and to investigate the root-reinforcing capability of AH Plus sealer. STUDY DESIGN: In groups 1-3 (n = 24) canals were instrumented with either GT files, FlexMaster, or stainless steel hand instruments. Twelve teeth from each group were obturated with lateral compaction using gutta-percha and AH Plus. Group 4 (n = 12) acted as uninstrumented controls. The force required to fracture the roots was measured. ANOVA, Scheffe, and t test were used for statistical analysis. RESULTS: The intact roots were significantly stronger than all groups with instrumented and unobturated roots (P < .05). Roots enlarged with GT files were significantly weaker (P < .05) than those instrumented with FlexMaster or hand instruments. Between the obturated and unobturated roots there were no significant differences (P > .05). CONCLUSIONS: The roots were significantly weakened by the preparation with greater taper instruments. An obturation with AH Plus did not increase the fracture resistance. PMID- 16360618 TI - Effect of limiting snack food variety on long-term sensory-specific satiety and monotony during obesity treatment. AB - Limiting meal variety decreases hedonic ratings of eaten foods more so than non eaten foods, demonstrating sensory-specific satiety. Exposure to a food over time decreases the food's hedonic ratings, indicating monotony. The effect of limiting food group variety over time on long-term sensory-specific satiety and monotony is unknown. Thirty overweight adults were randomized to one of two 8-week behavioral weight loss interventions. One condition limited snack food intake to one chosen snack food (reduced variety), while the other limited snack food intake to <1 serving/day (control), with no variety limit. In the reduced variety condition, hedonic ratings of the chosen snack food showed a decrease (p < .05) over time and decreased more (p < .05) than hedonic ratings of other snack foods. Weight loss (-7.4 +/- 5.8 lb) occurred in both conditions. Limiting food group variety over 8 weeks produced long-term sensory-specific satiety and monotony. Future research should examine if limiting food group variety over an extended time affects intake and could be used as a technique in weight loss interventions. PMID- 16360619 TI - Eating disorders and disordered eating among patients seeking non-surgical weight loss treatment in Sweden. AB - The aim of this cross-sectional descriptive study of 194 Swedish men and women seeking non-surgical weight-loss treatment was to investigate the presence of eating disorders and binge eating symptoms and to compare these two groups of patients with a group without eating disorder- or binge eating symptoms. The groups were compared in regard to co-morbid psychopathology, Health Related Quality of Life (HRQL) and anthropometric data. Of the total sample, 9.8% fulfilled criteria for any eating disorder. An additional 7.2% indicated binge eating symptoms without having an eating disorder. The three groups were significantly different in regard to psychopathology scales and most HRQL items. Eating disorders and binge eating symptoms are common among patients seeking non surgical weight-loss treatments in Sweden and both groups showed elevated levels of co-morbid psychopathology and lower HRQL compared to patients without disordered eating. These findings point to the importance of assessing the full range of eating disorder symptoms and disorders as well as HRQL and co-morbid psychopathology before weight-loss treatment, as these factors might affect treatment outcome. PMID- 16360620 TI - The specific content of core beliefs and schema in adolescent girls high and low in eating disorder symptoms. AB - The specific content of core beliefs and schema in adolescent girls high and low in eating disorder symptoms was investigated using the Eating Disorder Belief Questionnaire (EDBQ)-negative self-beliefs sub-scale and Young Schema Questionnaire (YSQ). Girls with high Eating Attitude Test (EAT) scores had higher scores on both measures (using total and sub-scale scores) than girls with low EAT scores. However, all scores were related to eating disorder as well as depressive symptoms. Analysis of individual items on the EDBQ and YSQ found that scores on all EDBQ, but not all YSQ items, differed between the two groups, with higher scores in the high EAT group. However, a number of individual items (on both measures) in the high EAT group, as well as in the low EAT group, were related specifically to eating disorder symptoms and not also to depressive symptoms. PMID- 16360621 TI - Is comorbidity in the eating disorders related to perceptions of parenting? Criterion validity of the revised Young Parenting Inventory. AB - BACKGROUND: Recent studies support the reliability and validity of the Young Parenting Inventory-Revised (YPI-R) and its use in investigating the role of parenting in the aetiology and maintenance of eating pathology. However, criterion validity has yet to be fully established. To investigate one aspect of criterion validity, this study examines the association between parenting and comorbid problems in the eating disorders (including general psychopathology and impulsivity). METHOD: The participants were 124 women with eating disorders. They completed the YPI-R and the Brief Symptom Inventory (BSI; a measure of general psychopathology). They were also interviewed about their use of a number of impulsive behaviours. RESULTS: YPI-R scales were significant predictors of one of the nine BSI scales, and distinguished those patients who did or did not use specific impulsive behaviours. DISCUSSION: The criterion validity of the YPI-R is partially supported with regards to general psychopathology and impulsivity. The findings highlight the specificity of the parenting styles measured by the YPI-R, and the need for further research using this tool. PMID- 16360622 TI - Gender and binge eating among bariatric surgery candidates. AB - Bariatric surgery is an increasingly popular treatment for severe obesity. However, while rates of obesity are comparable across genders, women are much more likely to undergo bariatric surgery. Little research exists on gender differences among bariatric surgery candidates. This study examined gender differences and their correlates within a sample of 487 patients presenting for bariatric surgery. Results indicated that women were more depressed than men, dieted more, and were more likely to report that their weight interfered with feeling good about themselves. There were no differences in BED rates or binge eating. Furthermore, among women, both depression and self-esteem were associated with binge eating symptomatology. However, among men, only depression contributed to binge eating scores. These findings provide evidence for the significance of binge eating and depression among both male and female bariatric surgery candidates. PMID- 16360623 TI - Disordered eating attitudes and symptomatology among adolescent girls in the United Arab Emirates. AB - OBJECTIVE: To ascertain the prevalence and correlates of disordered eating attitudes and symptomatology in a school-based Arab population. METHODS: A representative stratified random sample of 495 adolescent girls completed the Eating Attitudes Test (EAT-40) in the United Arab Emirates (UAE). A randomly selected subgroup from the those who scored above the recommended cut-off of 30 on the EAT-40 and an equal number of those who scored below 30 were interviewed in stage 2 by a psychiatrist, blind to the EAT scores, using the Schedule for Affective Disorders and Schizophrenia (KSADS) and DSM-IV criteria for the presence of eating disorders. RESULTS: 116 girls (23.4%) scored above the recommended cut-off on EAT. High EAT score was associated with age, BMI, internalization of thin ideal and drive for thinness, knowing someone on a weight loss strategy, having a family member with weight-related or mental health problem and watching western TV programs. Using clinical interview of 100 girls in stage 2, half of those who scored above the cut-off on EAT were found to have a propensity for anorexic behavior, while 2% met the criteria for the full clinical syndrome. CONCLUSIONS: Thin body preoccupation as well as family and social factors are important in the development of abnormal eating attitudes among adolescents in this Arabian Gulf country. PMID- 16360624 TI - Smoking status and weight loss in three weight loss programs. AB - The purpose of the study was to examine differences in current, former, and never smokers' weight loss in three comprehensive weight loss programs. Archival data from 389 overweight participants enrolled in three comprehensive weight loss programs were analyzed. The programs differed in length and by type of diet (food based low calorie diet, partial meal replacement low calorie diet, and supplement based very low calorie diet). Significant differences in weight loss were found among former, current, and never smokers in one weight loss program (partial meal replacement low calorie diet). Post hoc analyses showed that former smokers lost significantly more weight than current smokers. Results suggest that current smokers are not more successful at behavioral weight loss than former smokers, and if anything, are less successful. Former smokers do not appear to have difficulty losing weight compared to current and never smokers. PMID- 16360625 TI - Neuroticism and introversion: a risky combination for disordered eating among a non-clinical sample of undergraduate women. AB - Recent evidence suggests that people who score low on measures of sociability may be at risk for certain types of psychopathology, including mood and anxiety disorders. In an attempt to extend these findings to other forms of psychopathology, we examined levels of neuroticism and extraversion in relation to eating problems in a non-clinical sample of undergraduate women. The Eysenck Personality Questionnaire (EPQ), Eating Disorders Inventory (EDI), and the Eating Attitudes Test (EAT-26) were completed by 196 first-year undergraduate females. We found that high neuroticism was related to high scores on both of the EDI subscales (Bulimia and Drive for Thinness) as well as high scores on the EAT-26 measure, replicating previous work. In addition, neuroticism served as a moderator such that lower extraversion (i.e., introversion) was related to greater disordered eating, but only for those women who scored high on neuroticism. Thus, a combination of neuroticism and introversion may be a risk factor for symptoms of eating disorders in a non-clinical sample of university women. PMID- 16360626 TI - The relationship between eating self-efficacy and eating disorder symptoms in a non-clinical sample. AB - Most research on eating self-efficacy has focused on its relationship with eating behaviors and weight-loss in clinical populations. The purpose of this study was to investigate the relationship between eating self-efficacy and the behavioral and psychological characteristics associated with eating disorders in a non clinical sample of adults. A total of 219 men and women aged 18 and older completed questionnaires measuring eating disorder symptoms and eating self efficacy. The results indicated that low confidence in the ability to control eating while experiencing negative emotions was associated with greater weight preoccupation and bulimic thought and behaviors. In addition, low confidence in ability to control eating when an abundance of food is available was inversely related to feelings of ineffectiveness or general negative self-evaluation. Ultimately, the findings suggest that low eating self-efficacy may also be associated with eating problems within populations not seeking treatment for either eating disorders or weight-loss. The implications of the findings are discussed. PMID- 16360627 TI - Current indications for contrast echocardiography imaging. AB - The assessment of regional ventricular function is dependent on good endocardial definition. Suboptimal images can be converted to diagnostic recordings in the majority of patients by contrast agents, which have become an indispensable aid in rest and stress echocardiography. In particular for stress echocardiography image quality is essential and contrast administration is of great importance. However this diagnostic procedure must be performed following the indications which reflect the risks of the procedure and consider the benefits of an accurate diagnosis on further patient management. The contraindications recently introduced in the use of the echo-contrast agent SonoVue for acute cardiac patients reflect the same contraindications which have been applied in stress echocardiography for several years. Clinical trials and post-marketing surveillance have demonstrated that this approach is safe with no fatalities reported. For all ultrasound contrast media, side effects have been reported but they are usually mild. However, rare allergic and potentially life threatening reactions may occur and the investigators have to be prepared for such an event with appropriate drugs stored in the echo department. In a recent retrospective analysis in 751 consecutive stress echocardiograms the use of contrast during dobutamine stress echocardiography was not associated with an increased risk of side effects. The incidence of side effects was very low and different in patients receiving Optison, SonoVue or without contrast agent. CONCLUSION: Ultrasound contrast agents are licensed for improvement of endocardial border definition. Data from clinical trials and wide clinical experience indicate an excellent risk/benefit ratio if the current contraindications are applied. PMID- 16360628 TI - Quantification methods in contrast echocardiography. AB - New technologies and the availability of new echo-contrast agents have resulted in advances of diagnostic and prognostic indications of left ventricular opacification (LVO) and myocardial perfusion. The clinical diagnostic value of ultrasound contrast media for LVO and its impact on the clinical decision-making process has been demonstrated in several studies. Recent research aims at developing new quantitative software to improve the delineation of the endocardial border, to assess 3D myocardial perfusion for more accurate regional/global LV function measurements, and to evaluate 4D intra-cardiac flow dynamics. Furthermore, a general consensus has been reached on the incremental value of myocardial contrast echocardiography (MCE) for obtaining additional information in both chronic and acute coronary artery disease (CAD) patients and on the possibility to make quantitative measurements of microvascular damage. Q contrast is a new software system which provides quantitative measurements to generate parametric images of microcirculatory flow. In a research project including 120 patients, Q-contrast software has been tested to assess the role of contrast in AMI (Acute Myocardial Infarction Contrast Imaging (A.M.I.C.I. Study); good agreement between parametric MCE and SPECT has been found. Preliminary results further confirm that quantitative MCE may provide additional clinical value over qualitative information for the assessment of LV function and of the effects of coronary artery disease on the myocardial microcirculation (viability, ischemia or infarct). PMID- 16360629 TI - Clinical value and perspectives of contrast echocardiography. Concluding remarks and take home messages. Bracco symposium, Athens, Greece, 1-4 December 2004. PMID- 16360630 TI - Clinical benefits of contrast-enhanced echocardiography during rest and stress examinations. AB - Despite the recent introduction of tissue harmonic imaging in echocardiography, 10-15% of patients have poor endocardial border definition. This may lead to erroneous assessment of regional and global left ventricular (LV) function or to further diagnostic imaging with another modality thus increasing the costs for the healthcare system. The recent development of second generation contrast echocardiography agents such as SonoVue has resulted in several studies showing the value of these agents to outline endocardium clearly, thereby improving assessment of LV function. The use of these contrast agents has also opened the possibility of automated and quantitative LV function assessment, making it more accurate and reproducible. Other major clinical uses of these contrast agents are evaluation of LV masses such as thrombus and tumors, and better definition of LV structure such as delineating LV aneurysm, pseudoaneurysm; and non-compaction of LV and apical cardiomyopathy. Furthermore, the use of these contrast agents during stress not only improved the assessment of wall motion but also made possible the evaluation of myocardial perfusion, thereby increasing diagnostic accuracy for the detection of coronary artery disease. PMID- 16360631 TI - After the storm. PMID- 16360632 TI - Regional medicine following a large earthquake: economy-class syndrome. PMID- 16360633 TI - Request for volunteers. PMID- 16360634 TI - IMERT deployment to Baton Rouge, Louisiana in response to Hurricane Katrina, September 2005. AB - An Illinois volunteer emergency response team was deployed to assist in the disaster care efforts following Hurricane Katrina. The team joined local care providers to establish a field hospital on the campus of Louisiana State University in Baton Rouge, Louisiana. The team worked to support local providers, establish order, and assist more than 3000 evacuees. The challenges and lessons learned from this deployment are identified. PMID- 16360635 TI - Rapid assessment of the needs and health status in Santa Rosa and Escambia counties, Florida, after Hurricane Ivan, September 2004. AB - BACKGROUND: Hurricane Ivan, a Category 3 storm, devastated the Florida panhandle on September 16, 2004, causing extensive property damage and 24 deaths. The Florida Department of Health requested assistance from the Centers for Disease Control and Prevention to conduct a rapid assessment in Santa Rosa and Escambia counties to determine the health impact of the hurricane and needs of the affected population. METHODS: A questionnaire was administered 6 days after the hurricane made landfall. The survey instrument elicited information about house damage, illness/injury, and access to utilities. A modified cluster sampling method was used to select 30 clusters in each county. Seven households were interviewed in each cluster and reported weighted frequencies. RESULTS: Three quarters of houses in each county were damaged. Households in Santa Rosa and Escambia lacked basic utilities, including regular garbage pick-up, telephone service, and electricity. Fifty-four percent of households in Santa Rosa and 27% in Escambia reported using a generator. The most commonly self-reported health conditions were sleep disturbances in Santa Rosa (54%) and upper respiratory problems in Escambia (46%). Injuries were reported in fewer than 15% of households in each county. CONCLUSION: Rapid restoration of power, telephone services, and debris pick-up remained a priority 1 week after the event. Findings demonstrated the need for (1) mental health and primary care services, (2) information about safe generator use, and (3) ways to access medical care and medications. PMID- 16360636 TI - Development of a state medical surge plan, Part II: Components of a medical surge plan. AB - In 2003, the Utah State Department of Health received funding from the Health Resources and Services Administration to develop a medical surge plan to increase the number of available hospital beds in the state by 1250 beds, including 125 beds for burn or critical trauma patients. A prior article discussed the planning procedures and process. This article describes the major components of the plan, including analysis of threats, direction and control, activation and system response; communications; and critical issues. PMID- 16360637 TI - Venous thromboembolism in earthquake victims. AB - Sudden death from pulmonary thromboembolism following an earthquake has not been reported to date. This is a published case report of a 48-year-old woman who died suddenly of a pulmonary embolism on October 28, 2004. She lived in the Chuetu District in Japan, where a big epicentral earthquake occurred on October 23. She evacuated her house with her family and spent 4 of 5 nights in a private car. Until November 1, 8 persons who had been living out of their cars after evacuation from their houses in the District were treated for pulmonary thromboembolism. The immobility that comes with living in a car should be recognized as a risk factor for venous thromboembolism. In addition, potential preventive measures should be recommended, including provisions for emergency shelters, access to other temporary housing, public education, and the use of compression stockings. PMID- 16360638 TI - Case report: adapting the HEICS approach for flu vaccination clinic. PMID- 16360639 TI - Fast dual-excitation ratiometry with light-emitting diodes and high-speed liquid crystal shutters. AB - Dual-excitation ratiometric dyes permit quantitative measurements of Ca2+ concentrations ([Ca2+]s), by minimizing the effects of several artifacts that are unrelated to changes in [Ca2+]. These dyes are excited at two different wavelengths, and the resultant fluorescence intensities are measured sequentially. Therefore, it is difficult to follow fast [Ca2+] dynamics or [Ca2+] changes in highly motile cell samples. To overcome this problem, we have developed a new dual-excitation ratiometry system that employs two high-power light-emitting diodes (LEDs), two high-speed liquid crystal shutters, and a CCD camera. The open/close operation of the two shutters is synchronized with the on/off switching of the two LEDs. This system increases the rate at which ratio measurements are made to 1 kHz, and provides ratio images at 10-100 Hz depending on the signal intensity. We demonstrate the effectiveness of this system by monitoring changes in [Ca2+] in cardiac muscle cells loaded with Fura-2. PMID- 16360640 TI - Novel protein kinase interacts with the Cucumber mosaic virus 1a methyltransferase domain. AB - The Cucumber mosaic virus (CMV)-encoded 1a protein has been implicated to play a role in replication of the viral genome along with 2a and one or more host factors. To identify the host cell factors interacting with CMV 1a, we used the yeast two-hybrid system using tobacco cDNA library. One of the cDNA clones encoded a protein homologous to the Arabidopsis putative protein kinase and was designated as Tcoi2 (Tobacco CMV 1a interacting protein 2). Tcoi2 specifically interacted with methyltransferase (MT) domain of CMV 1a protein in yeast cell. In vitro analyses using recombinant proteins showed that Tcoi2 also specifically interacted with CMV 1a MT domain. Tcoi2 did not have autophosphorylation activity but phosphorylated CMV 1a MT domain. Analysis of the subcellular localization of the Tcoi2 fused to GFP demonstrated that it is targeted to the endoplasmic reticulum. These results suggest Tcoi2 as a novel host factor that is capable of interacting and phosphorylating MT domain of CMV 1a protein. PMID- 16360641 TI - Identification of extracellular signal-regulated kinase 3 as a new interaction partner of cyclin D3. AB - Cyclin D3, like cyclin D1 and D2 isoforms, is a crucial component of the core cell cycle machinery in mammalian cells. It also exhibits its unique properties in many other physiological processes. In the present study, using yeast two hybrid screening, we identified ERK3, an atypical mitogen-activated protein kinase (MAPK), as a cyclin D3 binding partner. GST pull-down assays showed that cyclin D3 interacts directly and specifically with ERK3 in vitro. The binding of cyclin D3 and ERK3 was further confirmed in vivo by co-immunoprecipitation assay and confocal microscopic analysis. Moreover, carboxy-terminal extension of ERK3 was responsible for its association with intact cyclin D3. These findings further expand distinct roles of cyclin D3 and suggest the potential activity of ERK3 in cell proliferation. PMID- 16360642 TI - Characterization of HCV-like particles produced in a human hepatoma cell line by a recombinant baculovirus. AB - Although processing of the hepatitis C virus (HCV) polyprotein and characterization of each of its viral proteins have been described in detail, analysis of the structure and assembly of HCV particles has been hampered by the lack of a robust cell culture system to support efficient replication of HCV. In this study, we generated HCV-like particles (HCV-LP) using a recombinant baculovirus encoding structural and a part of non-structural proteins in a human hepatoma cell line. The HCV-LP exhibited a buoyant density of 1.17 g/ml in CsCl equilibrium gradient and particles of 40 to 50 nm in diameter. Binding of the HCV LP to human hepatoma cells was partially inhibited by the treatment with anti hCD81 antibody, in contrast to the hCD81-independent binding of HCV-LP produced in insect cells. These results indicate that HCV-LP generated in different types of cells exhibit different cellular tropism for binding to target cells. PMID- 16360643 TI - Expression analysis of the fpr (ferredoxin-NADP+ reductase) gene in Pseudomonas putida KT2440. AB - The ferredoxin-NADP+ reductase (fpr) participates in cellular defense against oxidative damage. The fpr expression in Pseudomonas putida KT2440 is induced by oxidative and osmotic stresses. FinR, a LysR-type transcriptional factor near the fpr gene in the P. putida KT2440 genome, is required for induction of the fpr under both conditions. We have shown that the fpr and finR gene products can counteract the effects of oxidative and osmotic stresses. Interestingly, FinR independent expression occurs either during a long period of incubation with paraquat or with high concentrations of oxidative stress agent. This result indicates that there may be additional regulators present in the P. putida KT2440 genome. In contrast to in vivo expression kinetics of fpr from the plant pathogen, Pseudomonas syringae, the fpr gene from P. putida KT2440 exhibited unusually prolonged expression after oxidative stress. Transcriptional fusion and Northern blot analysis studies indicated that the FinR is negatively autoregulated. Expression of the fpr promoter was higher in minimal media than in rich media during exponential phase growth. Consistent with this result, the fpr and finR mutants had a long lag phase in minimal media in contrast to wild-type growth characteristics. Antioxidants such as ascorbate could increase the growth rate of all tested strains in minimal media. This result confirmed that P. putida KT2440 experienced more oxidative stress during exponential growth in minimal media than in rich media. Endogenous promoter activity of the fpr gene is much higher during exponential growth than during stationary growth. These findings demonstrate new relationships between fpr, finR, and the physiology of oxidative stress in P. putida KT2440. PMID- 16360644 TI - Inhibition of transcription factor NF-kappaB signaling proteins IKKbeta and p65 through specific cysteine residues by epoxyquinone A monomer: correlation with its anti-cancer cell growth activity. AB - Transcription factor NF-kappaB is constitutively active in many human chronic inflammatory diseases and cancers. Epoxyquinone A monomer (EqM), a synthetic derivative of the natural product epoxyquinol A, has previously been shown to be a potent inhibitor of tumor necrosis factor-alpha (TNF-alpha)-induced activation of NF-kappaB, but the mechanism by which EqM inhibits NF-kappaB activation was not known. In this report, we show that EqM blocks activation of NF-kappaB by inhibiting two molecular targets: IkappaB kinase IKKbeta and NF-kappaB subunit p65. EqM inhibits TNF-alpha-induced IkappaBalpha phosphorylation and degradation by targeting IKKbeta, and an alanine substitution for Cys179 in the activation loop of IKKbeta makes it resistant to EqM-mediated inhibition. EqM also directly inhibits DNA binding by p65, but not p50; moreover, replacement of Cys38 in p65 with Ser abolishes EqM-mediated inhibition of DNA binding. Pretreatment of cells with reducing agent dithiothreitol dose-dependently reduces EqM-mediated inhibition of NF-kappaB, further suggesting that EqM directly modifies the thiol group of Cys residues in protein targets. Modifications of the exocyclic alkene of EqM substantially reduce EqM's ability to inhibit NF-kappaB activation. In the human SUDHL-4 lymphoma cell line, EqM inhibits both proliferation and NF-kappaB DNA binding, and activates caspase-3 activity. EqM also effectively inhibits the growth of human leukemia, kidney, and colon cancer cell lines in the NCI's tumor cell panel. Among six colon cancer cell lines, those with low amounts of constitutive NF-kappaB DNA-binding activity are generally more sensitive to growth inhibition by EqM. Taken together, these results suggest that EqM inhibits growth and induces cell death in tumor cells through a mechanism that involves inhibition of NF-kappaB activity at multiple steps in the signaling pathway. PMID- 16360646 TI - Proximal HNF1 element is essential for the induction of human UDP glucuronosyltransferase 1A1 by glucocorticoid receptor. AB - Previous study showed noinduction of the reporter gene (-3174/+14) of UGT1A1 in HepG2 by bilirubin, but induction by dexamethasone (DEX). This induction was enhanced seven-fold by the co-expression of human glucocorticoid receptor (GR) and was inhibited by a GR antagonist, RU486, indicating stimulation by DEX-GR. Meanwhile, we could not detect stimulation by beta-estradiol, phenobarbital or rifampicin (RIF) in the presence of GR. We investigated the position playing a role in this induction by GR in the promoter region of UGT1A1 using deletion mutants, and clarified the essential sequence (-75/-63) for the binding site of hepatocyte nuclear factor 1 (HNF1). However, GR did not bind directly to this sequence, because UGT-PE2 did not compete for binding to a glucocorticoid responsive element (GRE) probe in an electrophoretic mobility shift assay (EMSA) method. Labeled [(32)P]DNA probe of HNF1 binds with nuclear extracts as shown by the EMSA. This shift of the complex of probe-protein was not inhibited by unlabeled GRE but was inhibited by unlabeled HNF1 element. This shift was not influenced by the addition of anti-GR, but was super-shifted by the addition of anti-HNF1. GR did not stimulate the induction of HNF1, because we detected no elevation of the mRNA level of HNF1 by reverse transcription-polymerase chain reaction (RT-PCR). Therefore, the induction of UGT1A1 by DEX-GR did not depend on the elevation of HNF1 but on the interaction of GR with HNF1 or the activation of HNF1 through the transcription of other proteins. Also given the lack of evidence of binding of DEX-GR to HNF1 in the EMSA, the data suggest that the mechanism of DEX-GRE effect on HNF1 is indirect by whatever mechanisms. PMID- 16360645 TI - Anti-tumour activity in non-small cell lung cancer models and toxicity profiles for novel ruthenium(II) based organo-metallic compounds. AB - Novel ruthenium(II) organo-metallic compounds are active in ovarian cancer models [Aird RE, Cummings J, Ritchie AA, Muir M, Morris RE, Chen H, et al. In vitro and in vivo activity and cross resistance profiles of novel ruthenium(II) organometallic arene complexes in human ovarian cancer. Br J Cancer 2002;86(10):1652-7]. [(eta6-C6H5C6H5)Ru(en)Cl]+ (as a PF6 salt, where en=ethylenediamine (RM175)) has been evaluated in a 13-cell line panel. Particular sensitivity (approximately 10-fold lower than mean IC50) was noted in breast cancer and non-small cell lung cancer cell lines. In addition, IC50 in the A549 was 2 microM and RM175 (25 mg kg-1, days 1 and 5, i.p.) caused a significant (p=0.004) growth delay in a xenograft model. HC11 [(eta6 tetrahydroanthracene)Ru(en)Cl]PF6 was more potent in the A549 cell line (IC50 0.5 microM). HC11 (25 mg kg-1, days 1, 8 and 15, i.p.) was also active in vivo. Following RM175 25 mg kg-1, days 1 and 5, and 15 mg kg-1, days 1-5, HC11 25 and 40 mg kg-1, day 1, elevated alanine transaminase levels were detected, suggesting hepatotoxicity. No changes were observed in kidney or haematological parameters. In liver sections, multi-focal hepatic necrosis was seen, becoming confluent at high doses of HC11. In vitro studies confirmed that HC11 was more toxic than RM175 to fresh human hepatocytes and equitoxic to mithramycin. Liver toxicity may be related to the arene ligand and modification may reduce the potential for hepatic toxicity, while maintaining the anti-tumour activity seen. PMID- 16360647 TI - The glutamate release inhibitor riluzole attenuates the formation of conditioned place aversion induced by naloxone in rats undergoing a single morphine exposure. AB - Acute morphine exposure has been hypothesized to produce long-lasting central changes that contribute to the withdrawal aversion. We have most recently demonstrated that those changes may involve the glutamatergic system, including multiple classes of receptors. The present study was undertaken to further determine the involvement of the glutamatergic system by examining the effect of riluzole, a glutamate release inhibitor, on the motivational component of withdrawal from acute morphine dependence. The role of the amygdala in the action of riluzole was also assessed. We investigated the effects of riluzole on the conditioned place aversion (CPA) induced by naloxone-precipitated withdrawal from a single morphine exposure 24 h before, and on c-Fos expression within the amygdala during the withdrawal period in rats. Riluzole (2, 4, 8 mg/kg) dose dependently attenuated the CPA without producing place conditioning itself. This result provided further evidence that glutamatergic mechanisms may be recruited in adaptational changes following acute morphine exposure and play a role in withdrawal aversion. In addition, riluzole appeared to produce nonspecific effects on c-Fos expression by itself, without specifically modifying c-Fos expression following naloxone-precipitated withdrawal in any region of the amygdala examined, suggesting that the amygdala may not serve as a specific site of action for riluzole. PMID- 16360648 TI - Ephrin/Eph receptor expression in brain of adult nonhuman primates: implications for neuroadaptation. AB - In developing brain, Eph receptors and their ephrin ligands (Ephs/ephrins) are implicated in facilitating topographic guidance of a number of pathways, including the nigrostriatal and mesolimbic dopamine (DA) pathways. In adult rodent brain, these molecules are implicated in neuronal plasticity associated with learning and memory. Cocaine significantly alters the expression of select members of this family of axonal guidance molecules, implicating Ephs, ephrins in drug-induced neuroadaptation. The potential contribution of Ephs, ephrins to cocaine-induced reorganization of striatal circuitry brain in primates [Saka, E., Goodrich, C., Harlan, P., Madras, B.K., Graybiel, A.M., 2004. Repetitive behaviors in monkeys are linked to specific striatal activation patterns. J. Neurosci. 24, 7557-7565] is unknown because there are no documented reports of Eph/ephrin expression or function in adult primate brain. We now report that brains of adult old and new world monkeys express mRNA encoding EphA4 receptor and ephrin-B2 ligand, implicated in topographic guidance of dopamine and striatal neurons during development. Their encoded proteins distributed highly selectively in regions of adult monkey brain. EphA4 mRNA levels were prominent in the DA-rich caudate/putamen, nucleus accumbens and globus pallidus, as well as the medial and orbitofrontal cortices, hippocampus, amygdala, thalamus and cerebellum. Immunocytochemical localization of EphA4 protein revealed discrete expression in caudate/putamen, globus pallidus, substantia nigra, cerebellar Purkinje cells, pyramidal cells of frontal cortices (layers II, III and V) and the subgranular zone of the hippocampus. Evidence for EphA4 expression in dopamine neurons emerged from colocalization with tyrosine-hydroxylase-positive terminals in striatum and substantia nigra and ventral tegmental area cell bodies. The association of axonal guidance molecules with drug-induced reorganization of adult primate brain circuitry warrants investigation. PMID- 16360649 TI - Neonatal pinealectomy induces Purkinje cell loss in the cerebellum of the chick: a stereological study. AB - Melatonin plays an important role in certain physiological functions and morphological features of various structures. In the current study, the effects of pinealectomy on Purkinje cell number and morphological features of developing cerebellum in the chick were investigated using stereological methods. Fifteen Hybro Broiler newly hatched chicks were divided into three groups: a pinealectomized group (n = 5), sham-operated group (n = 5) and a non pinealectomized control group (n = 5). Surgical pinealectomy was performed in 3 day-old chicks. In the 8th week, all animals were sacrificed for histopathological evaluation and subsequent stereological analysis. Each layer volume of molecular (+Purkinje cell), granular and white matter in the cerebellum was estimated in all animals. It was found that there was no significant difference for the volume of whole cerebellum and also molecular (+Purkinje cell) layer in these groups (P > 0.05). Nevertheless, the values of granular layer and white matter of sham-operated group were significantly different from those of control and pinealectomized animals (P < 0.01). It was also observed that pinealectomy significantly reduces the Purkinje cell number in cerebellar cortex (P < 0.01). The present study is the first stereological study to demonstrate the histomorphological effects of pinealectomy on the cerebellum in the chick. Our results suggest that pineal gland/melatonin might play an important role in morphological features of the developing cerebellum in the chick. PMID- 16360650 TI - Expression of secreted semaphorins and their receptors in specific neuromeres, boundaries, and neuronal groups in the developing mouse and chick brain. AB - Semaphorins constitute a family of signaling molecules with functions in axon pathfinding and neuronal migration. Neuropilins 1 and 2 have been identified as the ligand-binding component of semaphorin receptors. Both ligands and receptors are expressed in embryonic and adult organs in complementary and sometimes redundant patterns. In the present work, we compared the brain expression patterns of the class III semaphorins 3A, 3C, and 3F and neuropilins 1 and 2 between mouse and chick embryos at early developmental stages. Our studies revealed that expression of semaphorins is restricted in some cases to neuromeric transverse domains, to specific neuromeric boundaries, and to specific neuronal populations. Moreover, our studies also revealed coexpression of neuropilins and one or more semaphorins in some of the different expression sites. Comparison of the expression patterns between mouse and chick embryos showed large similarities, but important differences were also detected. PMID- 16360651 TI - Cryopreservation of cultured periosteum: effect of different cryoprotectants and pre-incubation protocols on cell viability and osteogenic potential. AB - Evidence has accumulated that periosteal cells have a great potential to regenerate bone. We have demonstrated that cultured periosteum (CP) in membrane form is an effective device to regenerate alveolar bone. To increase the availability of CP in a clinical environment, an effective cryopreservation protocol for CP has been developed. In this study, three different cryoprotectants (Me(2)SO, glycerol, and ethylene glycol) were used. The effect on cell viability of pre-incubation temperature, pre-incubation time, and agitation during incubation was investigated. Samples were stored at -196 degrees C for 10 days. Cell viability was assessed by a colorimetric cell viability assay using a tetrazolium salt, and the assay results were confirmed by confocal laser scanning microscopy after staining with a combination of calcein AM and ethidium homodimer 1. The activity of the cells after thawing was assessed by alkaline phosphatase assay. To assess the osteogenic potential of cryopreserved CP, the CP was grafted to calvarial defects in athymic rats. The greatest cell viability was obtained in the group equilibrated at 37 degrees C for 30 min with Me(2)SO, under agitation, showing 63.3 +/- 10.5% recovery. After cryopreservation, the cell growth of surviving cells was identical when Me(2)SO was used as a cryoprotectant. Alkaline phosphatase (ALP) activity was maintained in the groups cryopreserved with Me(2)SO and glycerol. The transplantation experiment showed that the calvarial defects were completely closed by grafting cryopreserved CP, which demonstrates that the osteogenic property of CP was well maintained. An efficient cryopreservation protocol for CP has been developed and this will provide a convenient and effective treatment option for bone regeneration in clinics. PMID- 16360652 TI - The function of neurofascin155 in oligodendrocytes is regulated by metalloprotease-mediated cleavage and ectodomain shedding. AB - Formation of the paranodal axo-glial junction requires the oligodendrocyte specific 155-kDa isoform of neurofascin (NF155). Here, we report the presence of two peptides in cultured oligodendrocytes, which are recognized by distinct NF155 specific antibodies and correspond to a membrane anchor of 30 kDa and a 125 kDa peptide, which is shed from the cells, indicating that it consists of the NF155 ectodomain. Transfection of OLN-93 cells with NF155 verified that both peptides originate from NF155 cleavage, and we present evidence that metalloproteases mediate NF155 processing. Interestingly, metalloprotease activity is required for NF155 transport into oligodendrocyte processes supporting the functional significance of NF155 cleavage. To further characterize NF155 cleavage and function, we transfected MDCK cells with NF155. Although ectodomain shedding was observed in polarized and non-polarized MDCK cells, surface localization of NF155 was restricted to the lateral membrane of polarized cells consistent with a role in cell-cell adhesion. Aggregation assays performed with OLN-93 cells confirmed that NF155 accelerates cell-cell adhesion in a metalloprotease-dependent manner. The physiological relevance of NF155 processing is corroborated by the presence of NF155 cleavage products in heavy myelin, suggesting a role of NF155 ectodomain shedding for the generation and/or stabilization of the nodal/paranodal architecture. PMID- 16360653 TI - Reduced expression of 1-cys peroxiredoxin in oxidative stress-induced cataracts. AB - 1-cys peroxiredoxin (1-cysPrx), a member of the peroxiredoxin family with a single conserved cysteine residue, reduces a broad spectrum of hydroperoxides. This study was undertaken to examine changes in 1-cysPrx expression in human cataract samples, human lens epithelial (HLE B3) cell line, and rat organ cultured lenses in response to oxidative insult induced by H2O2 or transforming growth factor-beta1 (TGF-beta1). Expression of 1-cysPrx mRNA and protein in HLE B3 cells increased in response to 2-8 ng ml(-1) TGF-beta1 and 50-75 microm H2O2 and then decreased below the control level at high doses (10 ng ml(-1) TGF-beta1 and 100-150 microm H2O2), as determined by Northern blot and immunoblot analysis. This reduction coincided with the decrease of cell viability. Immunoreactive 1 cysPrx protein was measured in capsulorrhexis specimens obtained from patients with anterior subcapsular cataract (ASC), nuclear sclerosis (NS), cortical spokes (CS), posterior subcapsular cataract (PSC), or white mature cataract (WC) at the time of cataract surgery. Significant reduction of 1-cysPrx protein was observed in ASC, PSC, and WC samples, but there was no statistical difference in CS and NS samples relative to normal control. Also, rat lens explants were cultured with 10 ng ml(-1) TGF-beta1 for approximately 5 days or 500 microm H2O2 for approximately 2 days. Subsequently, expression of 1-cysPrx mRNA and protein in the lens capsules was evaluated. Rat lens explants treated with TGF-beta1 or H2O2 developed a cataract similar to human ASC or WC, respectively, which resulted in a markedly decreased expression of 1-cysPrx mRNA and protein. Collectively, these findings show that expression patterns of 1-cysPrx gene in the lens are changed in response to oxidative stress, a major factor in the etiology of cataract. PMID- 16360654 TI - Treatment of experimental autoimmune uveoretinitis with poly(lactic acid) nanoparticles encapsulating betamethasone phosphate. AB - We have developed nanoparticles (NPs), which are capable of targeting a specific lesion and gradually releasing the agent at the site over a prolonged time period after a single intravenous administration. In this study, we evaluated the effects of intravenously administered poly(lactic acid) nanoparticles encapsulating betamethasone phosphate (BP-PLA NPs) on experimental autoimmune uveoretinitis (EAU) in Lewis rats. To determine the localization of NPs within the retina and choroid of rats with EAU, rhodamine (Rh)-encapsulated PLA NPs were injected intravenously and visualized by confocal microscopy. After the disease onset of EAU induced by S-antigen peptide in Lewis rats, either BP-PLA NPs, BP, or saline was injected intravenously, and the eyes were obtained 7 days following treatment and the histological score was determined. The clinical course of EAU was examined using pathological findings and the expression of the glial fibrillary acidic protein, rod opsin, and the surface markers of inflammatory cells (ED1 and pan T-cell) were immunohistochemically determined. Furthermore, T cell proliferation and delayed-type hypersensitivity (DTH) to S-antigen were assessed. Intravenously injected Rh-PLA NPs accumulated in the retina and choroid of rats with EAU within 3 hr and remained over the succeeding 7-day-period. Furthermore, systemically administered BP-PLA NPs reduced the clinical scores of rats with EAU in 1 day, which were maintained for 2 weeks and decreased the histological scores. In addition, the ocular infiltration of activated T-cells and macrophages in addition to the hypertrophy of Muller cells were markedly reduced with this treatment. Meanwhile, T-cell proliferation and DTH of BP-PLA NPs-treated rats against S-antigen peptide were not significantly different from those of saline-treated rats. Systemically administered BP-PLA NPs inhibit the development of EAU due to the targeting and the sustained release of steroids in situ. The results of these studies suggest that the systemic administration of BP PLA NPs may lead to a new therapeutic strategy in controlling intraocular inflammation. PMID- 16360655 TI - Injuries of the posterior cruciate ligament and posterolateral corner of the knee. AB - Injuries of the posterior cruciate ligament (PCL) and posterolateral corner (PLC) of the knee are less common than those of the anterior cruciate ligament (ACL) and their significance is often under-appreciated in the acute setting. Even when recognised, knowledge of the natural history and outcome of treatment has lagged behind that of the ACL and has led to confusion over the indications for operative treatment. Recent developments in the understanding of the anatomy and biomechanics of this area of the knee have led to improvements in management and a renewed interest in these potentially disabling injuries. The aim of this review is to bring the trauma generalist abreast of these recent developments and to improve diagnosis through a heightened index of suspicion and use of appropriate special investigations. The principles of management of both isolated and combined injuries to the PCL and PLC, in the acute and chronic settings, are described. PMID- 16360656 TI - A phenomenographic study of registered nurses' understanding of their role in student learning--an Australian perspective. AB - BACKGROUND: Students may be 'buddied' with registered nurses during their clinical experience since the designated clinical facilitator cannot be available for each student at all times. Little is known about the way registered nurses understand this informal role. OBJECTIVES: The rationale for this study was to gain an insight of the variation of understanding registered nurses have of their role with students, and explored the qualitatively different ways registered nurses perceive their role with students on clinical experience and the implications of this understanding for student learning. DESIGN: A phenomenographic approach was used to identify the variation of understanding and meaning of the role of the registered nurse with students on clinical practice from the perspective of the registered nurse. Phenomenography is a field of descriptive research concerned with the variation in ways people experience and understand similar phenomena. SETTINGS AND PARTICIPANTS: A purposive sample of 30 registered nurses from 15 public and private hospitals in central and south eastern Queensland, Australia. METHODS: Individual semi-structured interviews from a final sample of 28 interviews were analysed to identify Categories of Description. RESULTS: Eight variations of understanding registered nurses have of their informal role with students were identified. The registered nurses' understanding varies from a focus that is 'student-centred', to 'completion of workload-centred', to 'registered nurse control', to a preference for no contact with students. As a consequence some students may have positive learning experiences while others will have limited learning opportunities. CONCLUSIONS: The research highlights the varied ways registered nurses understand their role with students that may promote or impede the quality of student learning and development to meet professional competency standards. Formal recognition of the complexity of the registered nurse role by health care agencies and tertiary education providers is essential to ensure registered nurses have adequate preparation for their role with students. PMID- 16360657 TI - Activation of serotonergic 5-HT(1A) receptors in the lateral parabrachial nucleus increases NaCl intake. AB - Previous studies using non-specific serotonergic agonists and antagonists have shown the importance of serotonergic inhibitory mechanisms in the lateral parabrachial nucleus (LPBN) for controlling sodium and water intake. In the present study, we investigated whether the serotonergic 5-HT(1A) receptor subtype in the LPBN participates in this control. Male Holtzman rats had cannulas implanted bilaterally into the LPBN. Bilateral injections of the 5-HT(1A) receptor agonist, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT, 0.1, 1.25, and 2.5 microg/0.2 microl), into the LPBN enhanced 0.3 M NaCl and water intake of rats injected subcutaneously with the diuretic furosemide (10 mg/kg bw) and a low dose of the angiotensin-converting enzyme inhibitor, captopril (5 mg/kg bw). The increase in NaCl intake produced by 8-OH-DPAT injections was reduced in dose related manner by pre-treating the LPBN with the selective 5-HT(1A) serotonergic antagonist, WAY-100635 (WAY, 1 and 2 microg/0.2 microl). In contrast, WAY did not affect water intake produced by 8-OH-DPAT. WAY-100635 injected alone into the LPBN had no effect on NaCl ingestion. Injections of 8-OH-DAPT (0.1 microg/0.2 microl) into the LPBN also increased 0.3 M NaCl intake induced by 24-h sodium depletion (furosemide, 20 mg/kg bw plus 24 h of sodium-free diet). Serotonin (5 HT, 20 mug/0.2 mul) injected alone or combined with 8-OH-DPAT into the LPBN reduced 24-h sodium depletion-induced 0.3 M NaCl intake. Therefore, the activation of serotonergic 5-HT(1A) receptors in the LPBN increases stimulated hypertonic NaCl and water intake, while 5-HT injections into the LPBN reduce NaCl intake and prevent the effects of serotonergic 5-HT(1A) receptor activation. PMID- 16360658 TI - Prostaglandin EP4 receptor agonist protects against acute neurotoxicity. AB - Under various abnormal physiologic conditions, overactivation of glutamate-gated ion channel receptor family members, including NMDA receptors, causes increase in COX-2 expression and generation of prostaglandins. PGE(2) exerts its physiologic actions mainly through its PGE(2) prostanoid (EP) receptors. In the present study, the role of the EP4 receptor against NMDA-induced excitotoxicity was investigated. Using the EP4 receptor agonist ONO-AE1-329, which has relative selectivity toward murine EP receptors on the order of EP1:EP2:EP3:EP4 of >1000:210:120:1, respectively, we questioned whether activation of the EP4 receptors has the potential to attenuate injury in brain. Mice were pretreated by intracerebroventricular injection with different doses of ONO-AE1-329 (0.1, 1, and 10 nmol; n = 9/group) and, after 20 min, by a single unilateral intrastriatal injection of NMDA (15 nmol, n = 12). NMDA injection produced a significant lesion in the ipsilateral striatum. This lesion volume was significantly reduced in groups that were pretreated with ONO-AE1-329, with maximum protection of more than 32% at 10 nmol. This is the first study revealing the protective effect of ONO-AE1-329 in an acute model of excitotoxicity in brain, and it suggests that preferential stimulation of EP4 receptors attenuates excitotoxic brain injury. PMID- 16360659 TI - Endothelin-1 as a central mediator of LPS-induced fever in rats. AB - Fever induced by E. coli lipopolysaccharide (LPS) in rats is substantially reduced by blockade of central endothelin ET(B) receptors. This study explores the role of endothelin-1 as a central mediator of fever in rats, by investigating the effect of a pyrogenic dose of LPS on the levels of big endothelin-1 and endothelin-1 in the cerebrospinal fluid (CSF) and endothelin-1 in the plasma. We further assessed whether the increase in body temperature caused by central injection of endothelin-1 constitutes solely a hyperthermia or a true integrated febrile response. LPS (5 mug kg(-1), i.v.) induced fever which peaked at 1.16 +/- 0.24 degrees C within 2 h and remained stable up to 5 h. CSF levels of immunoreactive (ir) big endothelin-1 decreased to undetectable levels at 3 h after LPS, returning only partially at 5 h post-injection. CSF ir-endothelin-1 levels were undetectable in saline-treated animals, but reached 21.9 +/- 5.2 fmol ml(-1) at 3 h after LPS treatment. Plasma ir-endothelin-1 levels were unchanged after saline or LPS. Central injection of endothelin-1 (1 pmol, i.c.v.) caused long-lasting increases in body temperature (0.81 +/- 0.17 degrees C, 3 h), but simultaneously decreased tail skin temperature (-1.10 +/- 0.26 degrees C), indicating cutaneous vasoconstriction. Moreover, endothelin-1 induced fever (1.0 +/- 0.3 degrees C, 3 h) when injected into the preoptic area of the anterior hypothalamus (100 fmol), but not i.v. (1 or 10 pmol). These data suggest that endothelin-1 is produced in the brain and acts centrally as a mediator of LPS induced fever. PMID- 16360660 TI - Analysis of polymeric phenolics in red wines using different techniques combined with gel permeation chromatography fractionation. AB - A multiple-step analytical method was developed to improve the analysis of polymeric phenolics in red wines. With a common initial step based on the fractionation of wine phenolics by gel permeation chromatography (GPC), different analytical techniques were used: high-performance liquid chromatography-diode array detection (HPLC-DAD), HPLC-mass spectrometry (MS), capillary zone electrophoresis (CZE) and spectrophotometry. This method proved to be valid for analyzing different families of phenolic compounds, such as monomeric phenolics and their derivatives, polymeric pigments and proanthocyanidins. The analytical characteristics of fractionation by GPC were studied and the method was fully validated, yielding satisfactory statistical results. GPC fractionation substantially improved the analysis of polymeric pigments by CZE, in terms of response, repeatability and reproducibility. It also represented an improvement in the traditional vanillin assay used for proanthocyanidin (PA) quantification. Astringent proanthocyanidins were also analyzed using a simple combined method that allowed these compounds, for which only general indexes were available, to be quantified. PMID- 16360661 TI - Data-directed scan sequence for the general assignment of C-glycosylflavone O glycosides in plant extracts by liquid chromatography-ion trap mass spectrometry. AB - An ion trap LC-MS/MS method is described for the analysis of C-glycosylflavone O glycosides in crude methanolic extracts of plants. The method employs survey scans with and without the application of up-front collision induced dissociation (CID) to generate diagnostic ions for data-directed MS/MS. The spectra acquired allow assignment of the C-linked sugar to either the C-6 or C-8 position of the aglycone and provide data on the molecular mass of the compound, the number and type of O-linked sugars and the molecular mass of the flavone aglycone. These data for the majority of C-glycosylflavone O-glycosides in an extract are obtained automatically in one LC-MS/MS analysis without manual pre-programming. Key to the assignment of the C-6 or C-8 site of C-glycosylation is the generation, by up-front CID, of the (0,1)X+ product ion formed by internal cleavage of the C-linked sugar. MS/MS of this ion is found to have diagnostic value in addition to the (0,2)X+ product ion described by other authors. Ion trap MS/MS spectra of [M+H]+ of the 6,8-di-C-glycosylflavones schaftoside and isoschaftoside show an additional and previously unreported diagnostic product ion that is useful in determining the type of sugar at the C-6 position. The product ion spectra of protonated kaempferol 3-O-glucosylrhamnosides show similarities to the spectra of C-glycosylflavone O-glycosides; this is a potential source of confusion if the analysis of such glycosides is limited solely to MS/MS of [M+H]+. PMID- 16360662 TI - Faster axial band dispersion in a monolithic silica column than in a particle packed column. AB - The contribution of molecular diffusion to peak broadening was studied in a reversed-phase HPLC system, consisting of a monolithic silica C18 column and methanol-water mobile phase. Study on the band broadening effect of holding a solute in a column or elution at very low linear velocity of mobile phase allowed facile determination of the contribution of the molecular diffusion term. Less obstruction against molecular diffusion, or the faster axial band dispersion in a monolithic silica column than in a particle-packed column, was found both in mobile phase and in stationary phase. PMID- 16360663 TI - Size fractionation and characterization of natural colloids by flow-field flow fractionation coupled to multi-angle laser light scattering. AB - Flow-field flow fractionation (FlFFF) coupled to multi-angle laser light scattering (MALLS) was evaluated for size and shape determination of standard spherical and arbitrarily shaped natural colloids. Different fitting methods for light scattering data retrieved from MALLS were evaluated to determine the particle size of spherical standards and natural colloids. In addition, FlFFF was optimized for best fractionation in connection to MALLS, minimal colloids membrane interaction, and minimal sample losses. FlFFF, calibrated with standard particles, was used to determine hydrodynamic diameter, or radius (D(h) or R(h)), of the fractionated colloids, whereas the MALLS was used to determine root mean square radius of gyration (R(g)) for fractionated colloids. Combining both results, by calculating the R(g)/R(h) ratio, allows an estimation of colloid deviation from the shape of homogeneous sphere. Accordingly, this study demonstrates that, FlFFF-MALLS is a valuable technique for characterizing heterogeneous and arbitrarily shaped natural colloidal particles in terms of size and shape. To check the usefulness of FlFFF-MALLS in natural colloid studies, the technique was used to investigate the sedimentation behavior of extracted soil colloidal particles. Results illustrate that, in a silty till sample, carbonates function as cement between the colloidal particles, and consequently, change their sedimentation behavior. On the other hand, carbonate dissolution generates a more homogeneous colloidal sample. PMID- 16360664 TI - Liquid-liquid-liquid microextraction with automated movement of the acceptor and the donor phase for the extraction of phenoxyacetic acids prior to liquid chromatography detection. AB - A simple liquid-liquid-liquid microextraction with automated movement of the acceptor and the donor phase (LLLME/AMADP) technique is described for the quantitative determination of five phenoxyacetic acids in water using a disposable and ready to use hollow fiber. The target compounds were extracted from the acidified sample solution (donor phase) into the organic solvent residing in the pores of the hollow fiber and then back extracted into the alkaline solution (acceptor phase) inside the lumen of the hollow fiber. The fiber was held by a conventional 10-microl syringe. The acceptor phase was sandwiched between the plunger and a small volume of the organic solvent (microcap). The acceptor solution was repeatedly moved in and out of the hollow fiber assisted by a programmable syringe pump. This repeated movement provides a fresh acceptor phase to come in-contact with the organic phase and thus enhancing extraction kinetics leading to high enrichment of the analytes. The microcap separates the aqueous acceptor phase and the donor phase in addition of being partially responsible for mass transfer of the analytes from donor solution (moving in and out of the hollow fiber from the open end of the fiber) to the acceptor solution. Separation and quantitative analyses were then performed using liquid chromatography (LC) with ultraviolet (UV) detection at 280 nm. Various parameters affecting the extraction efficiency viz. type of organic solvent used for immobilization in the pores of the hollow fiber, extraction time, stirring speed, effect of sodium chloride, and concentration of donor and acceptor phases were studied. Repeatability (RSD, 3.2-7.4%), correlation coefficient (0.996 0.999), detection limit (0.2-2.8 ng ml(-1)) and enrichment factors (129-240) were also investigated. Relative recovery (87-101%) and absolute recoveries (4.6-13%) have also been calculated. The developed method was applied for the analysis of river water. PMID- 16360665 TI - A validated higher-performance liquid chromatography method for quantification of cinchonain Ib in bark and phytopharmaceuticals of Trichilia catigua used as Catuaba. AB - The hydroalcoholic extract, prepared from authentic chopped barks of Trichilia catigua, was evaluated by high-performance liquid chromatography using a diode array detector (200-400 mn). The crude extract was purified by rotation locular counter-current chromatography and the chloroform fraction obtained was clean-up by solid-phase extraction. With the aim of getting preliminary structure information on-line, the methanol fraction thus obtained was analyzed by gradient elution using the diode array detector coupled to a mass spectrometer. The presence of flavalignan in this extract was inferred by the chromatographic band, in the total ion current trace, that had an [M-H](-) = 451. With this information, cinchonain Ib was isolated as a pure compound from the crude hydroalcoholic extract using a solid-phase extraction procedure for the sample clean-up followed by a semi-preparative separation using the reverse mode of elution. The isolated compound, after complete characterization, was used as an external standard for the development and validation of a method for the analysis of this compound in herbal medicines using the ultraviolet as the detector. The validated method has been successfully applied for quantification of cinchonain Ib in commercialized herbal medicines sold as Catuaba in Brazil and also in standard chopped barks of T. catigua. PMID- 16360666 TI - N-Menthoxycarbonylation combined with trimethylsilylation for enantioseparation of beta-blockers by achiral dual-column gas chromatography. AB - Solvent extractive two-phase menthoxycarbonyl (MnOC) derivatization was combined with trimethylsilyl (TMS) reaction for enantioseparation of beta-blockers by gas chromatography employing achiral DB-5 and DB-17 dual-columns of different polarity. beta-Blockers in alkaline solution were vortex-mixed with menthyl chloroformate present in dichloromethane to be extracted as diastereomeric N-MnOC derivatives. The subsequent O(N)-TMS reaction allowed complete enantioseparations of two beta-blockers and partial separations of five as N-MnOC/O(N)-TMS derivatives in a single analysis. The temperature-programmed retention index sets were characteristic of each derivative, facilitating chiral discrimination of each enantiomer. PMID- 16360667 TI - On the mechanisms of dissolution of montroydite [HgO(s)]: dependence of the dissolution rate on pH, temperature, and stirring rate. AB - The dissolution behavior of montroydite (HgO) has been studied using a fully automated system. Dissolution data under equilibrium conditions are in agreement with previously published data and indicate that HgO solubility is relatively high and constant between pH 4 and 10.1 and increases markedly at pH<4. The dissolution rate also has similar behavior: it is relatively high and constant between pH 4 and 10.1 and increases sharply at pH<4. The dissolution process obeys a three-dimensional contraction or attrition mechanism. The dissolution rate increases with increasing temperature and stirring rate and is the result of mixed transport and reaction control. The rate of HgO dissolution is considerably higher than that of other divalent metal oxides at low pH. This high rate is due to the ability of Hg(II) to rapidly exchange its ligands. Data suggest that montroydite will only occur in nature in highly contaminated sites and indicate that Hg oxidation products that are formed at the liquid Hg/water interface may dissolve rapidly. PMID- 16360668 TI - Effect of powder oxide content on the expansion and stability of PM-route Al foams. AB - The oxide content in Al powders has been found to have a significant effect on the expansion and stability of foams made via a PM route. With low oxide contents (O<0.3 wt%) expansion is moderate and the foam structure is unstable. Larger expansions, improved foam stability and more homogeneous foam structures are achieved if the amount of oxide in the powder is moderate (O=0.3-0.6 wt%). Foaming precursors with excessive oxide contents (O>0.6 wt%) results in small expansions but very stable foam structures. Oxides were observed to form clusters of crumpled films, which at higher levels form a network which restricts the drainage of liquid from the cell walls and Plateau borders, but which also inhibit foam expansion. Oxide clusters in the cell walls lead to a decrease in the minimum cell wall thickness, resulting in an increase in foam expansion. PMID- 16360669 TI - Acoustic-perceptual correlates of voice quality in elderly men and women. AB - Common perceptual characteristics of the elderly voice include hoarseness, breathiness, instability, and a change in the pitch of the voice. Although research is available concerning changes in the elderly voice, little research has been completed to examine the relationship between the perception of voice quality and acoustic measures. The purpose of this study was to examine the relationship between perceived breathiness and hoarseness and selected acoustic variables. The results of the study revealed significant age-related differences in fundamental frequency standard deviation, amplitude perturbation quotient, and noise-to-harmonic ratio. No significant difference in perceived hoarseness was found between the young and elderly speakers. However, the young women were perceived as significantly more breathy than the elderly women. Moderate correlations were found between the perceptions of hoarseness and breathiness and the acoustic measures. These findings are discussed relative to age-related laryngeal changes that may contribute to the perception of aging. LEARNING OUTCOMES: (1) To understand the perceptual attributes of the aging voice in men and women. (2) To understand the relation between the perception of voice quality and acoustic measures of voice production in the aging voice. PMID- 16360670 TI - Visual cortex: overcoming a no-go for plasticity. AB - Normally, the brain can be shaped by sensory experience only during a so-called critical period early in life. Recent research has shed light on the factors determining the end of the critical period, and on how cortical plasticity might be re-established in adulthood. PMID- 16360671 TI - Axis formation: squint comes into focus. AB - Gene products provided by the mother to the embryo determine the body axes in most animals. A recent study in zebrafish proposes that the TGFss signal Squint is one such factor. PMID- 16360672 TI - Figs and fig wasps. PMID- 16360673 TI - Schizophrenia. PMID- 16360674 TI - The cause of colouration in the ctenophore Beroe cucumis. PMID- 16360675 TI - Circadian pathway: the other shoe drops. AB - Three new papers report the long-awaited functional characterization of the Drosophila receptor for the circadian-rhythm-regulating signalling molecule PIGMENT DISPERSING FACTOR (PDF). The discovery of the PDF receptor heralds progress in understanding the circadian pacemaker circuit and output pathways in insects. PMID- 16360676 TI - DNA sliding clamps: just the right twist to load onto DNA. AB - Two recent papers illuminate a key step in DNA sliding clamp loading: one reveals the structure of the PCNA clamp wrapped around DNA--still open from being loaded- while the other finds that the clamp may assist this process by forming a right handed helix upon opening. PMID- 16360677 TI - Coevolution: the geographic mosaic of coevolutionary arms races. AB - Coevolutionary arms races between species can favor exaggeration of traits for attack and defense, but relentless escalation of these arms races does not necessarily occur in all populations. PMID- 16360678 TI - Yeast polarity: negative feedback shifts the focus. AB - A new study of Cdc42p polarization in yeast suggests that the actin cytoskeleton can destabilize the polarity axis, causing Cdc42p foci to wander aimlessly around the cell cortex. PMID- 16360679 TI - Olfactory coding: inhibition reshapes odor responses. AB - Olfactory information is dramatically restructured as it makes its way through the brain. Recent work using a remarkable experimental preparation has revealed how this transformation is achieved. PMID- 16360680 TI - Cytokinesis: breaking the ties that bind. AB - It has been unclear how cells complete cell division and resolve membrane connections to bring about cell separation. Recent work has shown that targeted secretion to the midbody is required to complete cell division. PMID- 16360681 TI - Cerebellar complex spike firing is suitable to induce as well as to stabilize motor learning. AB - BACKGROUND: Cerebellar Purkinje cells (PC) generate two responses: the simple spike (SS), with high firing rates (>100 Hz), and the complex spike (CS), characterized by conspicuously low discharge rates (1-2 Hz). Contemporary theories of cerebellar learning suggest that the CS discharge pattern encodes an error signal that drives changes in SS activity, ultimately related to motor behavior. This then predicts that CS will discharge in relation to the error and at random once the error has been nulled by the new behavior. RESULTS: We tested this hypothesis with saccadic adaptation in macaque monkeys as a model of cerebellar-dependent motor learning. During saccadic adaptation, error information unconsciously changes the endpoint of a saccade prompted by a visual target that shifts its final position during the saccade. We recorded CS from PC of the posterior vermis before, during, and after saccadic adaptation. In clear contradiction to the "error signal" concept, we found that CS occurred at random before adaptation onset, i.e., when the error was maximal, and built up to a specific saccade-related discharge profile during the course of adaptation. This profile became most pronounced at the end of adaptation, i.e., when the error had been nulled. CONCLUSIONS: We suggest that CS firing may underlie the stabilization of a learned motor behavior, rather than serving as an electrophysiological correlate of an error. PMID- 16360682 TI - Swe1p responds to cytoskeletal perturbation, not bud size, in S. cerevisiae. AB - BACKGROUND: S. cerevisiae cells must grow to a critical size in G1 in order to pass start and enter the cell cycle. A recent study proposed that in addition to the mother size control in G1, the bud must grow to a critical bud size in G2 in order to enter mitosis. Insufficient bud size would cause G2 arrest enforced by the mitotic inhibitor Swe1p, explaining previous findings that some perturbations that block bud growth also trigger Swe1p-dependent cell-cycle arrest. RESULTS: We tested the critical-bud-size hypothesis. We found that halting bud growth by inactivation of the myosin Myo2p did not trigger Swe1p-dependent arrest in budded cells, even when the buds were very small. Moreover, Swe1p did not affect cell cycle progression in unstressed cells, even when bud size was decreased by overriding G1 size control. Actin depolymerization did cause Swe1p-dependent arrest in small-budded but not large-budded cells, as previously reported. However, we found that the key determinant of cell-cycle arrest in those circumstances was not bud size, but rather the relative abundance of the Swe1p mitotic inhibitor and the mitosis-promoting cyclins. CONCLUSIONS: Swe1p does not respond to insufficient bud size. Instead, actin stress empowers Swe1p to promote arrest. The effectiveness of Swe1p in promoting that arrest declines as cells progress through the cell cycle. PMID- 16360683 TI - Nonmuscle myosin II generates forces that transmit tension and drive contraction in multiple tissues during dorsal closure. AB - BACKGROUND: The morphogenic movements that characterize embryonic development require the precise temporal and spatial control of cell-shape changes. Drosophila dorsal closure is a well-established model for epithelial sheet morphogenesis, and mutations in more than 60 genes cause defects in closure. Closure requires that four forces, derived from distinct tissues, be precisely balanced. The proteins responsible for generating each of the forces have not been determined. RESULTS: We document dorsal closure in living embryos to show that mutations in nonmuscle myosin II (encoded by zipper; zip/MyoII) disrupt the integrity of multiple tissues during closure. We demonstrate that MyoII localization is distinct from, but overlaps, F-actin in the supracellular purse string, whereas in the amnioserosa and lateral epidermis each has similar, cortical distributions. In zip/MyoII mutant embryos, we restore MyoII function either ubiquitously or specifically in the leading edge, amnioserosa, or lateral epidermis and find that zip/MyoII function in any one tissue can rescue closure. Using a novel, transgenic mosaic approach, we establish that contractility of the supracellular purse string in leading-edge cells requires zip/MyoII-generated forces; that zip/MyoII function is responsible for the apical contraction of amnioserosa cells; that zip/MyoII is important for zipping; and that defects in zip/MyoII contractility cause the misalignment of the lateral-epidermal sheets during seam formation. CONCLUSIONS: We establish that zip/MyoII is responsible for generating the forces that drive cell-shape changes in each of the force generating tissues that contribute to closure. This highly conserved contractile protein likely drives cell-sheet movements throughout phylogeny. PMID- 16360684 TI - The egg surface LDL receptor repeat-containing proteins EGG-1 and EGG-2 are required for fertilization in Caenorhabditis elegans. AB - The molecular machinery that mediates sperm-egg interactions at fertilization is largely unknown. We identify two partially redundant egg surface LDL receptor repeat-containing proteins (EGG-1 and EGG-2) that are required for Caenorhabditis elegans fertility in hermaphrodites, but not males. Wild-type sperm cannot enter the morphologically normal oocytes produced by hermaphrodites that lack egg-1 and egg-2 function despite direct gamete contact. Furthermore, we find that levels of meiotic maturation/ovulation and sperm migratory behavior are altered in egg-1 mutants. These observations suggest an unexpected regulatory link between fertilization and other events necessary for reproductive success. egg-1 and egg 2 are the result of a gene duplication in the nematode lineage leading to C. elegans. The two closely related species C. briggsae and C. remanei encode only a single egg-1/egg-2 homolog that is required for hermaphrodite/female fertility. In addition to being the first identified egg components of the nematode fertilization machinery, the egg-1 and egg-2 gene duplication could be vital with regards to maximizing C. elegans fecundity and understanding the evolutionary differentiation of molecular function and speciation. PMID- 16360685 TI - Exposure to host resistance mechanisms drives evolution of bacterial virulence in plants. AB - Bacterial pathogenicity to plants and animals has evolved through an arms race of attack and defense. Key players are bacterial effector proteins, which are delivered through the type III secretion system and suppress basal defenses . In plants, varietal resistance to disease is based on recognition of effectors by the products of resistance (R) genes . When recognized, the effector or in this scenario, avirulence (Avr) protein triggers the hypersensitive resistance reaction (HR), which generates antimicrobial conditions . Unfortunately, such gene-for-gene-based resistance commonly fails because of the emergence of virulent strains of the pathogen that no longer trigger the HR . We have followed the emergence of a new virulent pathotype of the halo-blight pathogen Pseudomonas syringae pv. phaseolicola within leaves of a resistant variety of bean. Exposure to the HR led to the selection of strains lacking the avirulence (effector) gene avrPphB (or hopAR1), which triggers defense in varieties with the matching R3 resistance gene. Loss of avrPphB was through deletion of a 106 kb genomic island (PPHGI-1) that shares features with integrative and conjugative elements (ICElands) and also pathogenicity islands (PAIs) in diverse bacteria . We provide a molecular explanation of how exposure to resistance mechanisms in plants drives the evolution of new virulent forms of pathogens. PMID- 16360686 TI - Interaction between EB1 and p150glued is required for anaphase astral microtubule elongation and stimulation of cytokinesis. AB - In animal cells, microtubules (MTs) of the mitotic apparatus (MA) communicate with the cell cortex to stimulate cytokinesis; however, the molecular nature of this stimulus remains elusive . A signal for cytokinesis likely involves the MT plus end binding family of proteins, which includes EB1, p150glued, APC, LIS1, and CLIP-170. These proteins modulate MT dynamics and facilitate interactions between growing MTs and their intracellular targets, including kinetochores, organelles, and the cell cortex . The dynein-dynactin complex mediates many of these microtubule capture events . We report that EB1 and p150glued interactions are required for stimulation of cytokinesis in dividing sea urchin eggs. Injected antibodies against EB1 or p150glued suppressed furrow ingression but did not prevent elongation of anaphase astral MTs toward the cortex, suggesting that EB1 and dynactin are both required for communication between the MA and the cortex. Targeted disruption of the interaction between EB1 and p150glued suppressed anaphase astral MT elongation and resulted in a delay of cytokinesis that could not be overcome by manipulation of the asters toward the cortex. We conclude that EB1 and dynactin participate in stimulation of the cleavage furrow, and their interaction promotes elongation of astral MTs at anaphase onset. PMID- 16360687 TI - The neural basis of the behavioral face-inversion effect. AB - Two of the most robust markers for "special" face processing are the behavioral face-inversion effect (FIE)-the disproportionate drop in recognition of upside down (inverted) stimuli relative to upright faces-and the face-selective fMRI response in the fusiform face area (FFA). However, the relationship between these two face-selective markers is unknown. Here we report that the behavioral FIE is closely associated with the fMRI response in the FFA, but not in other face selective or object-selective regions. The FFA and the face-selective region in the superior temporal sulcus (f_STS), but not the occipital face-selective region (OFA), showed a higher response to upright than inverted faces. However, only in the FFA was this fMRI-FIE positively correlated across subjects with the behavioral FIE. Second, the FFA, but not the f_STS, showed greater neural sensitivity to differences between faces when they were upright than inverted, suggesting a possible neural mechanism for the behavioral FIE. Although a similar trend was found in the occipital face area (OFA), it was less robust than the FFA. Taken together, our data suggest that among the face-selective and object selective regions, the FFA is a primary neural source of the behavioral FIE. PMID- 16360688 TI - Control of Shugoshin function during fission-yeast meiosis. AB - Meiosis consists of a single round of DNA replication followed by two consecutive nuclear divisions. During the first division (MI), sister kinetochores must orient toward the same pole to favor reductional segregation. Correct chromosome segregation during the second division (MII) requires the retention of centromeric cohesion until anaphase II. The spindle checkpoint protein Bub1 is essential for both processes in fission yeast . When bub1 is deleted, the Shugoshin protein Sgo1 is not recruited to centromeres, cohesin Rec8 does not persist at centromeres, and sister-chromatid cohesion is lost by the end of MI. Deletion of bub1 also affects kinetochore orientation because sister centromeres can move to opposite spindle poles in approximately 30% of MI divisions. We show here that these two functions are separable within the Bub1 protein. The N terminus of Bub1 is necessary and sufficient for Sgo1 targeting to centromeres and the protection of cohesion, whereas the C-terminal kinase domain acts together with Sgo2, the second fission-yeast Shugoshin protein, to promote sister kinetochore co-orientation during MI. Additional analyses suggest that the protection of centromeric cohesion does not operate when sister kinetochores attach to opposite spindle poles during MI. Sgo1-mediated protection of centromere cohesion might therefore be regulated by the mode of kinetochore attachment. PMID- 16360689 TI - Slit and Robo control cardiac cell polarity and morphogenesis. AB - Basic aspects of heart morphogenesis involving migration, cell polarization, tissue alignment, and lumen formation may be conserved between Drosophila and humans, but little is known about the mechanisms that orchestrate the assembly of the heart tube in either organism. The extracellular-matrix molecule Slit and its Robo-family receptors are conserved regulators of axonal guidance. Here, we report a novel role of the Drosophila slit, robo, and robo2 genes in heart morphogenesis. Slit and Robo proteins specifically accumulate at the dorsal midline between the bilateral myocardial progenitors forming a linear tube. Manipulation of Slit localization or its overexpression causes disruption in heart tube alignment and assembly, and slit-deficient hearts show disruptions in cell-polarity marker localization within the myocardium. Similar phenotypes are observed when Robo and Robo2 are manipulated. Rescue experiments suggest that Slit is secreted from the myocardial progenitors and that Robo and Robo2 act in myocardial and pericardial cells, respectively. Genetic interactions suggest a cardiac morphogenesis network involving Slit/Robo, cell-polarity proteins, and other membrane-associated proteins. We conclude that Slit and Robo proteins contribute significantly to Drosophila heart morphogenesis by guiding heart cell alignment and adhesion and/or by inhibiting cell mixing between the bilateral compartments of heart cell progenitors and ensuring proper polarity of the myocardial epithelium. PMID- 16360690 TI - Light activation of channelrhodopsin-2 in excitable cells of Caenorhabditis elegans triggers rapid behavioral responses. AB - For studying the function of specific neurons in their native circuitry, it is desired to precisely control their activity. This often requires dissection to allow accurate electrical stimulation or neurotransmitter application , and it is thus inherently difficult in live animals, especially in small model organisms. Here, we employed channelrhodopsin-2 (ChR2), a directly light-gated cation channel from the green alga Chlamydomonas reinhardtii, in excitable cells of the nematode Caenorhabditis elegans, to trigger specific behaviors, simply by illumination. Channelrhodopsins are 7-transmembrane-helix proteins that resemble the light-driven proton pump bacteriorhodopsin , and they also utilize the chromophore all-trans retinal, but to open an intrinsic cation pore. In muscle cells, light-activated ChR2 evoked strong, simultaneous contractions, which were reduced in the background of mutated L-type, voltage-gated Ca2+-channels (VGCCs) and ryanodine receptors (RyRs). Electrophysiological analysis demonstrated rapid inward currents that persisted as long as the illumination. When ChR2 was expressed in mechanosensory neurons, light evoked withdrawal behaviors that are normally elicited by mechanical stimulation. Furthermore, ChR2 enabled activity of these neurons in mutants lacking the MEC-4/MEC-10 mechanosensory ion channel . Thus, specific neurons or muscles expressing ChR2 can be quickly and reversibly activated by light in live and behaving, as well as dissected, animals. PMID- 16360691 TI - Guest Editorial: Intersections for mutual success in nursing and health services research. PMID- 16360697 TI - The intersection of nursing and health services research: overview of an agenda setting conference. PMID- 16360698 TI - Access in health services research: the battle of the frameworks. AB - BACKGROUND: Access is an important concept in the study of the organization, financing and delivery of healthcare services. It is also an important political symbol and policy goal. PURPOSE: This article reviews the major theoretical frameworks that have been used to support the study of access and measure the degree to which healthcare systems have met standards of access. METHOD: The article uses a critical review of the major works of the leading theorists in the field of access studies. DISCUSSION: Theories of access accept that it is a dynamic process where there is the potential for individuals and families to learn and modify their behavior. That learning and adaptation is less often explored in empirical research of access to health care. CONCLUSION: Researchers should consider the more dynamic aspects of access as they attempt to understand how to improve the health care delivery system. Access models can be used to direct the formulation of better health policy if they reflect real world processes. PMID- 16360699 TI - Health services research and the nursing workforce: access and utilization issues. AB - Access to personal health services is only possible if there are an adequate number and distribution of qualified nurses. However, most work on access has been conceptualized as access to physician services or particular types of settings such as hospital care, not as access to nurses. In this paper I have used the Institute of Medicine Access framework to conceptualize the relationship of the nursing workforce to several aspects of access, and evaluated both the current state of research and existing gaps. Some of the gaps identified in the research that need to be included in a health services research agenda for nursing are: (1) the effect of regional market factors (such as physician ratios and poverty), or attitudes on the supply of nurses; (2) the lack of a critical analysis of staffing and outcome research, and measures of utilization and access specific to nurses in a variety of settings; (3) the need for more and better data to evaluate nurse career patterns, nursing workforce educational capacity, and effectiveness; and (4) relevant workforce policies. PMID- 16360700 TI - The consequences of job stress for nurses' health: time for a check-up. AB - The processes and outcomes of nurses' work are described extensively in studies about patient care, nursing education and training, job satisfaction, health care quality and management, and organizational behavior. These studies evaluate the relationship between nurses' behavior and organizational health (ie, productivity) or between nurses' behavior and patient health (ie, medical error). Fewer studies probe the association between the nature of nursing work and the status of nurses' health despite the logical connection between how well nurses feel and how well they perform, or even, whether they discontinue working altogether for health reasons. Yet, for many nurses working in today's health care environment, work is a stressful part of their lives. This article explores the connections between stressful work and nurses' health, especially given the restructuring of their work in the current health care system. The working conditions that give rise to stress and the potential health consequences from it are well described in the general stress literature and summarized herein. Moreover, studies about nurses' work and nurses' health are discussed in light of the limitations for connecting job stress to job changes or health outcomes over time. Current approaches for dealing with nurses' stress, such as the attraction to "Magnetism", may inadvertently impede progress in this area. Recommendations for the future are included. PMID- 16360701 TI - Quality of care in nursing home organizations: establishing a health services research agenda. AB - Health services research has led to exciting new findings about the critical importance of the amount and type of nursing staff in nursing homes for improving the processes and outcomes of care. This paper reviews recent theoretical and research issues and outlines areas where research is needed. The nursing home research agenda for the future needs to concentrate on: (1) the relationship between structural measures of nursing (eg, staffing levels, education, turnover rates) and the outcomes and processes of care; (2) adequate processes of care and ways to improve the reliability of clinical outcome measures; (3) better ways to risk-adjust for resident characteristics; (4) the impact of nursing home characteristics (eg, ownership) and public policies (eg, reimbursement) on structural factors, processes, and outcomes; and (5) cost-effectiveness studies of nursing care at the organizational or system level. PMID- 16360702 TI - The cost and cost-effectiveness of nursing services in health care. AB - This article examines the literature on cost-effectiveness in nursing, and considers the relationship between this literature and decision-making in health care systems. Researchers have attempted to examine costs and benefits of nurse staffing and nursing interventions for decades. There are strong literatures for some topics, such as advanced practice nursing, clinical practices, occupational health nursing, and workplace training. However, there are gaps in the literature on the cost-effectiveness of nurse staffing patterns, the use of agency personnel, and changes in organizational structure. A review of 6 major health care management textbooks finds few references to cost-effectiveness analysis, suggesting that health care leaders have little education regarding how to conduct or evaluate economic studies. The agenda for nursing research on cost effectiveness is daunting. Research must be based on large, representative samples; provide clear, compelling results; discuss the importance of both costs and benefits in decision-making; and be published in highly-visible journals. PMID- 16360703 TI - The changing nature of the measurement of the economic impact of nursing care on health care organizations. AB - This paper adapts the perspective of organizational contingency theory to consider the changing nature of how the economic impact of nursing care upon health care organizations is measured. It is argued that useful measures of the economic impact of nursing care are a function of environmental, organizational, and technological circumstances. The increasing and diverse demands of health care consumers (environmental), the dramatic restructuring and re-engineering of the health care delivery system (organizational), and recent developments in the capabilities of and insights from information measurement practices (technological), have all provided opportunities for more meaningful measurement of the contributions of nursing care to the economic well-being of health care organizations. PMID- 16360704 TI - The contribution of organization theory to nursing health services research. AB - We review nursing and health services research on health care organizations over the period 1950 through 2004 to reveal the contribution of nursing to this field. Notwithstanding this rich tradition and the unique perspective of nursing researchers grounded in patient care production processes, the following gaps in nursing research remain: (1) the lack of theoretical frameworks about organizational factors relating to internal work processes; (2) the need for sophisticated methodologies to guide empirical investigations; (3) the difficulty in understanding how organizations adapt models for patient care delivery in response to market forces; (4) the paucity of attention to the impact of new technologies on the organization of patient care work processes. Given nurses' deep understanding of the inner workings of health care facilities, we hope to see an increasing number of research programs that tackle these deficiencies. PMID- 16360705 TI - The intersection of nursing and health services research: an agenda to guide future research. PMID- 16360706 TI - A review of performance-based measures of functional living skills. AB - There is growing recognition of the need to consider the effect of mental health treatments on functional independence/abilities, in addition to any effects on primary symptoms. A variety of performance-based measures of functional living skills have appeared in the medical literature over the past 25 years in discipline-specific journals, but there has been little attempt to synthesize this information to guide mental health clinicians and researchers in choosing among the available measures. Thus, the goal of the present review was (1) to identify and evaluate the strengths and weaknesses of currently available performance-based assessments of functional living skills for adult populations, and (2) to make specific recommendations regarding use of currently available instruments as well as recommendations for future instrument development. Through a comprehensive search of electronic citation databases for English-language articles on performance-based instruments, as well as via cross-referenced articles in the bibliography of each initially identified article, we identified 94 relevant articles covering 31 instruments, including 20 multi-domain instrument batteries, and 11 additional domain-specific performance-based instruments. Most identified instruments provided evidence of adequate reliability, but empirical data regarding the predictive validity of each scale in terms of actual "real world functioning" was less prevalent. Several good performance-based measures have been developed for individuals with dementia, psychiatric illnesses, and older healthy individuals. Future instrument development may benefit from a focus on brevity, empirical item selection as determined by patients/caregivers, predictive validity with an emphasis on change in functioning over time, and the relationship of performance to levels of competency required for the completion of everyday tasks. PMID- 16360707 TI - Phenylsulphonyl urenyl chalcone derivatives as dual inhibitors of cyclo-oxygenase 2 and 5-lipoxygenase. AB - Two series of phenylsulphonyl urenyl chalcone derivatives (UCH) with various patterns of substitution were tested for their effects on nitric oxide (NO) and prostaglandin E2 (PGE2) overproduction in RAW 264.7 macrophages. None of the tested compounds reduced NO production more than 50% at 10 microM but most of them inhibited the generation of PGE2 with IC50 values under the micromolar range. Me-UCH 1, Me-UCH 5, Me-UCH 9, Cl-UCH 1, and Cl-UCH 9 were selected to evaluate their influence on human leukocyte functions and eicosanoids generation. These derivatives selectively inhibited cyclo-oxygenase-2 (COX-2) activity in human monocytes being Me-UCH 5 the most potent (IC50 0.06 microM). Selected compounds also reduced leukotriene B4 synthesis in human neutrophils by a direct inhibition of 5-lipoxygenase (5-LO) activity, with IC50 values from 0.5 to 0.8 microM. In addition, lysosomal enzyme secretion, such as elastase or myeloperoxidase as well as superoxide generation in human neutrophils were also reduced in a similar range. Our findings indicate that UCH derivatives exert a dual inhibitory effect on COX-2/5-LO activity. The profile and potency of these compounds may have relevance for the modulation of the inflammatory and nociceptive responses with reduction of undesirable side-effects associated with NSAIDs. PMID- 16360708 TI - Effect of the difference in vehicles on gene expression in the rat liver- analysis of the control data in the Toxicogenomics Project Database. AB - The Toxicogenomics Project is a 5-year collaborative project by the Japanese government and pharmaceutical companies in 2002. Its aim is to construct a large scale toxicology database of 150 compounds orally administered to rats. The test consists of a single administration test (3, 6, 9 and 24 h) and a repeated administration test (3, 7, 14 and 28 days), and the conventional toxicology data together with the gene expression data in liver as analyzed by using Affymetrix GeneChip are being accumulated. In the project, either methylcellulose or corn oil is employed as vehicle. We examined whether the vehicle itself affects the analysis of gene expression and found that corn oil alone affected the food consumption and biochemical parameters mainly related to lipid metabolism, and this accompanied typical changes in the gene expression. Most of the genes modulated by corn oil were related to cholesterol or fatty acid metabolism (e.g., CYP7A1, CYP8B1, 3-hydroxy-3-methylglutaryl-Coenzyme A reductase, squalene epoxidase, angiopoietin-like protein 4, fatty acid synthase, fatty acid binding proteins), suggesting that the response was physiologic to the oil intake. Many of the lipid-related genes showed circadian rhythm within a day, but the expression pattern of general clock genes (e.g., period 2, arylhydrocarbon nuclear receptor translocator-like, D site albumin promoter binding protein) were unaffected by corn oil, suggesting that the effects are specific for lipid metabolism. These results would be useful for usage of the database especially when drugs with different vehicle control are compared. PMID- 16360709 TI - Changes of cerebral blood oxygenation and optical pathlength during activation and deactivation in the prefrontal cortex measured by time-resolved near infrared spectroscopy. AB - To determine the alterations in optical characteristics and cerebral blood oxygenation (CBO) during activation and deactivation, we evaluated the changes in mean optical pathlength (MOP) and CBO induced by a verbal fluency task (VFT) and driving simulation in the right and left prefrontal cortex (PFC), employing a newly developed time-resolved near infrared spectroscopy, which allows quantitative measurements of the evoked-CBO changes by determining the MOP with a sampling time of 1 s. The results demonstrated differences in MOP in the foreheads with the subjects and wavelength; however, there was no significant difference between the right and left foreheads (p > 0.05). Also, both the VFT and driving simulation task did not affect the MOP significantly as compared to that before the tasks (p > 0.05). In the bilateral PFCs, the VFT caused increases of oxyhemoglobin and total hemoglobin associated with a decrease of deoxyhemoglobin, while the driving simulation task caused decreases of oxyhemoglobin and total hemoglobin associated with an increase of deoxyhemoglobin; there were no significant differences in evoked-CBO changes between the right and left PFC. The present results will be useful for quantitative measurement of hemodynamic changes during activation and deactivation in the adults by near infrared spectroscopy. PMID- 16360710 TI - Retinal eccentricity effects on reaction time to imagined stimuli. AB - To cast light on the possible neural substrate of visual imagery we tested normal participants and one hemianopic patient on simple reaction time (RT) to real and imagined visual stimuli. In one experiment participants were to detect as quickly as possible a luminous square presented at one out of two different retinal eccentricities. A well known effect with visual stimuli is that RT is slower for peripheral versus central stimuli. We found that imagined stimuli showed an eccentricity effect similar to that obtained with real stimuli. However, this was not the case in a patient with a hemianopic visual field loss (quadrantanopia) as a result of damage to the optic radiation. Even though the patient showed no difficulty in imaging stimuli in the affected hemifield she did not show an eccentricity effect as was the case in her intact side. In a second experiment, normal participants showed faster RT to stimuli of larger size with either real or imagined stimuli. Overall, these results show that visual perception and imagination share a similar visuotopic organisation that is disrupted following deafferentation of the visual cortex. PMID- 16360711 TI - Caprylic acid infusion acts in the liver to decrease food intake in rats. AB - Hepatic portal vein (HPV) infusion of the medium chain fatty acid caprylic acid (CA; 2.3 mg/min, 40 microl/min) for 90 min beginning at dark onset in 18-h food deprived male rats reduced the size of the first nocturnal meal about 40% (P < 0.01) and reduced 24-h food intake by about 15% (P < 0.001). Identical infusions into the vena cava affected neither initial meal size nor food intake. HPV CA infusion attenuated the postprandial decreases in plasma free fatty acids (P < 0.01) and beta-hydroxybutyrate (P < 0.01). HPV CA infusions did not significantly reduce nocturnal saccharine intake in a two-bottle conditioned taste aversion test, and there was no association between the saccharine intake on the test day and the feeding-inhibitory effect of CA on the conditioning day. HPV CA infusion did not affect plasma concentrations of corticosterone or of the pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha. HPV CA infusion did not increase plasma concentration of the liver enzyme alanine aminotransferase, but did increase plasma concentration of gamma-glutamyl transferase, although not into the pathophysiological range. These data indicate that CA acts in the liver to produce a signal that inhibits feeding and that this inhibitory effect may be related to increases in hepatic fatty acid oxidation rather than be the result of aversion or toxicity. PMID- 16360712 TI - Butyl benzyl phthalate blocks Ca2+ signaling coupled with purinoceptor in rat PC12 cells. AB - Butyl benzyl phthalate (BBP) is a plasticizer and causes public concern because of its genomic estrogenic effects via estrogen receptors. We previously found that BBP has non-genomic effects, exerting inhibitory effects on the functional activities of nicotinic acetylcholine receptors (nAChR) in bovine adrenal chromaffin cells. nAChR belongs to the superfamily of neurotransmitter-gated channels, so does P2X purinoceptor that is widely distributed in the nervous system and play a role in pain reactions. In this study, we investigated the effects of BBP on the change of [Ca2+]c (cytosolic calcium ion concentration) under the stimulation of purinoceptors in PC12 cells and found that BBP inhibited ATP-induced [Ca2+]c rise (IC50=8.3 microM). The inhibitory rate of BBP remained under the increase of ATP concentration; therefore, the possibility of competitive inhibition was excluded. The inhibition of BBP on P2Y was excluded because its inhibition on ATP-induced [Ca2+]c rise was not found in the absence of extracellular Ca2+. BBP might have some actions on voltage-operated Ca2+ channels (VOCCs) since BBP inhibited the Ca2+ signaling responding to high K+ stimulation (IC50=1.2 microM). We suggest that BBP inhibits the ATP-induced [Ca2+]c rise via its non-competitive inhibition on P2X purinoceptors and VOCCs in the plasma membrane. PMID- 16360713 TI - Moving towards better health: a survey of transport authorities and primary care trusts in South West England. AB - OBJECTIVE: Transport, the way we travel and our ability to access amenities, is an important determinant of health. A survey was conducted to: (1) identify past and present joint working arrangements between transport authorities and health authorities; (2) understand the role and impact of transport themes in health improvement programmes (HIMPs) and health themes in local transport plans (LTPs); and (3) raise awareness of transport and health issues. STUDY DESIGN: A questionnaire survey of directors of public health of primary care trusts (PCTs) and local authority transport managers was undertaken in the South West of England, a region of 5 million people with 32 PCTs and 15 transport authorities. RESULTS: All the transport authorities and 66% of PCTs responded. Consultation with the health authority on the LTP had been full in 67% of transport authority areas and more limited in the remainder. Common targets in the LTP and HIMP had been agreed in 33% of transport authority areas, shared programmes or themes in 40%, and a single shared theme in 13%. LTP and HIMP shared themes included walking and cycling initiatives, road safety, school travel and rural access. LTP themes identified as best practice to deliver health gain included walking initiatives, rural access, school travel and road safety. Ongoing links with PCTs were in place in 73% of the transport areas. Of the PCTs that responded, 33% reported that they were working on a travel plan. CONCLUSIONS: The responses in this survey identified strengths and concerns in the way that health and transport issues are handled locally and are reflected in the LTPs and HIMPs. Recommendations were produced by public health and transport managers in government office South West to improve partnership working. PMID- 16360714 TI - Tideswell health survey: a population survey of the health needs and service utilization of a farming community. AB - OBJECTIVES: In the UK, there have been no epidemiological studies focusing on the general health status of rural communities, despite the prevalence of rural deprivation and the widely acknowledged link between poverty and ill health. The primary objective of this study was to test the null hypothesis that no differences existed between the health status of the farming and non-farming populations registered with one general practice, in order to inform a local health needs assessment. The secondary objective was to test the suitability of the research method as a model for a wider study. STUDY DESIGN: A postal questionnaire survey was undertaken with the total farming population (n = 248) registered with Tideswell general practice, and matched with an equivalent random sample of non-farmers (n = 248) from the remainder of the practice list. The main outcome measures were income, general health status, quality of life (EQ5D), musculoskeletal problems, mental health status and service utilization. RESULTS: Significant differences were found between the two groups. Farmers were identified as having health needs that were greater than those found in the local non-farming sample. In addition, the observed EQ5D value for farmers was found to be significantly lower than that found in social classes IV and V (0.78 vs 0.82) and the UK mean. CONCLUSIONS: This modest controlled study is the first of its kind in the UK. Health inequalities and inequities were identified that may be generalizable to other UK hill farmers. However, a larger, wider epidemiological study across different types of farming groups is now required to gain a fuller understanding of the health status of this indigenous rural population. The methodology employed in this study was successful and could serve as a model for such a study. PMID- 16360715 TI - Introduction: chemotherapies in the treatment of breast cancer. PMID- 16360716 TI - Microtubule targeting agents: basic mechanisms of multidrug resistance (MDR). AB - Multidrug resistance (MDR) results in the loss of activity of anticancer agents against cells possessing the MDR phenotype. In some cases, tumor cells are intrinsically resistant to chemotherapeutic drugs (intrinsic resistance), while in other instances cells become resistant after exposure to cytotoxic agents (acquired resistance). MDR has a significant impact on the management of patients with solid tumors, including those with breast cancer, where it has been previously correlated with a decreased response to treatment and a poor prognosis. Taxanes are among the most important components of chemotherapy regimens to treat recurrent breast cancer. In taxane-refractory cells, the two mechanisms most commonly associated with the development of MDR are the overexpression of members of the ATP-binding cassette family of transporters, of which P-glycoprotein is the best known, and alterations of tubulin, which is the cellular target of the taxanes. In vitro and in vivo models have been developed to study the development of MDR and to assess the potential clinical application of MDR modulators. However, despite promising advances using MDR modulators in preclinical models, clinical applications have met with limited success. Novel strategies are needed to circumvent the development of drug resistance. New cytotoxic agents capable of evading MDR offer hope for patients with breast cancer. PMID- 16360717 TI - Multidrug resistance/P-glycoprotein and breast cancer: review and meta-analysis. AB - Previously untreated breast cancer is relatively sensitive to a range of anticancer drugs. However, exposure to these drugs is often followed by acquisition of multidrug resistance, which is associated with a significantly worse outcome. One of the more widely studied mechanisms of drug resistance is the function of P-glycoprotein (P-gp), a membrane transporter with a wide range of substrates, including several anticancer agents, and a member of the ATP binding cassette superfamily of proteins. A review of the published literature indicates that P-gp expression is detected in a significant percentage of breast cancers. Moreover, P-gp expression is increased after exposure to chemotherapeutic drugs (particularly those known to be P-gp substrates), and correlates with a worse response to treatment, especially when detected following treatment, in both the adjuvant and neoadjuvant settings. Consequently, P-gp represents a potential biomarker of drug resistance. However, a direct role of P gp as a cause of clinical drug resistance has not been adequately tested in breast cancer. Future studies aimed at validating the mechanistic role of P-gp should include trials of multidrug resistance reversal using P-gp-specific inhibitors and relating results to the levels of P-gp expression. Future studies should also take into account the potentially multifactorial nature of multidrug resistance. PMID- 16360718 TI - Clinical management of recurrent breast cancer: development of multidrug resistance (MDR) and strategies to circumvent it. AB - The multidrug resistance (MDR) phenotype is often associated with recurrent breast cancer. Many cytotoxic agents used to treat breast cancer, such as anthracyclines and taxanes, are susceptible to MDR-mediated loss of sensitivity to these agents. Overexpression of mdr-1/P-glycoprotein (P-gp) is one of the main mechanisms underlying the development of the MDR phenotype. Also involved in the development of the MDR phenotype are other proteins from the ATP-binding cassette family of transporters (eg, MRP, BCRP), as well as alterations of tumor targets and their downstream effector molecules. Additionally, P-gp expression in other anatomic locations (such as the brush border of the gastrointestinal epithelium and blood-brain barrier) may further compromise the success of treatment for patients with breast cancer. Several strategies have been developed to overcome or circumvent MDR, mostly through inhibition or modulation of P-gp. Despite successful proof of concept in the laboratory, to date none of these agents has had a major impact in the clinic. PMID- 16360719 TI - Development of novel chemotherapeutic agents to evade the mechanisms of multidrug resistance (MDR). AB - A number of novel microtubule-targeting agents are currently under investigation. These agents can potentially evade the mechanisms underlying the development of the multidrug resistance (MDR) phenotype commonly associated with recurrent breast cancer. Epothilones are among the most advanced of the new agents in clinical development. Structurally unrelated to taxanes, epothilones may be poor substrates for MDR, and the expression of MDR proteins is not altered in epothilone-resistant in vitro models. Cross resistance between epothilones and taxanes is not observed in vitro or in vivo. Ixabepilone (BMS-247550) is a semisynthetic analog of epothilone B that has shown antitumor activity both in vitro and in vivo, including taxane-resistant human tumor xenograft models. Ixabepilone is currently being studied in phase III trials in patients with metastatic breast cancer as monotherapy and in combination with capecitabine. Activity has also been observed in other solid tumors. Patupilone (EPO906, epothilone B) and epothilone D (KOS-862) are in early phase I/II clinical studies in patients with a variety of solid tumors. The development of these novel agents may evade MDR and may improve the outcome of patients with breast cancer. PMID- 16360720 TI - Synthesis of multiply 13C-labeled furofuran lignans using 13C-labeled cinnamyl alcohols as building blocks. AB - Plant lignans are currently being widely studied for their potential benefits for human health as their consumption has been correlated with lower risks for developing chronic diseases, such as breast cancer and coronary heart disease. However, studies of some classes of lignans, in particular the furofurans, are hampered by the lack of suitable standards to allow accurate analysis. Herein, we report the syntheses of two racemic (13)C-labeled furofuran lignans [7,8,9 (13)C(3)]medioresinol and [7,8,9-(13)C(3)]sesamin as internal standards for LC-MS analysis. The labeled furofuran lignans were constructed from triply labeled cinnamyl alcohols, using a radical cyclization method. PMID- 16360721 TI - High affinity 17alpha-substituted estradiol derivatives: synthesis and evaluation of estrogen receptor agonist activity. AB - We synthesized four derivatives of 17beta-estradiol (E2) with an azide substitution on a 17alpha-side chain of varying length, namely 17alpha (azidopropargyl)-3,17beta-estradiol (5), its 17beta-azido derivative (diazide 7), 17alpha-(5-azido-pent-1-ynyl)-3,17beta-estradiol (6) and 17alpha-(azidopentyn-2 yl)-3,17beta-estradiol (10). While most of the derivatives had low (7) or marginal (6 and 10) relative binding affinity (RBA) for both types of estrogen receptor (ERalpha and ERbeta), the RBAalpha and RBAbeta of 5 were practically identical to those of E2. The estrogenic activity of the derivatives was assessed using estrogen-responsive breast (MCF-7) and endometrial cancer (Ishikawa) cells. While 5 was a potent and effective inducer of alkaline phosphatase in Ishikawa cells and 7 was less potent but as effective as 5, 6 was marginally active and 10 was totally inactive in this respect. In the presence of 0.1 nM E2, however, 6 exhibited some ER antagonist activity at the highest concentration tested (1 microM). Similar results were obtained as regards the potency and efficacy of stimulation of MCF-7 cell proliferation and induction of luciferase gene expression in MCF-7:D5L cells, a clone stably transfected with an estrogen responsive form of the gene. These data suggest that, while 5, 6, 7 and 10 interact with either type of ER in isolation, only 5 and 7 exhibit substantial ER agonist activity in the different estrogen-target cells examined, which could provide for photoaffinity labelling of the receptor in the cell as well as in isolation. PMID- 16360722 TI - Sulfation of tibolone metabolites by human postmenopausal liver and small intestinal sulfotransferases (SULTs). AB - Sulfation is a major pathway in humans for the biotransformation of steroid hormones and structurally related therapeutic agents. Tibolone is a synthetic steroid used for the treatment for climacteric symptoms and postmenopausal osteoporosis. Sulfation inactivates the hydroxylated metabolites, 3alpha hydroxytibolone (3alpha-OH-tibolone) and 3beta-hydroxytibolone (3beta-OH tibolone), and contributes to the regulation of tissue responses to tibolone. We detected SULT1A1, SULT1A3, SULT1E1 and SULT2A1 mRNA expression by RT-PCR in postmenopausal liver and small intestine. Liver pool (n=5) SULT activities measured with tibolone substrates reflected COS-1 expressed SULT2A1 and SULT1E1 activities. Liver SULT2A1 activity (1.8 +/- 0.3 units/mg protein, n = 8, mean +/- SEM), and activities with 3alpha-OH-tibolone (0.6 +/- 0.1, n = 8) and 3beta-OH tibolone (0.9 +/- 0.2, n = 8) were higher than SULT1E1 activities (<0.05, n = 10). SULT1E1 activities were low or not detected in many samples. Mean small intestinal activities were 0.03 +/- 0.01 with 3alpha-OH-tibolone and 0.04 +/- 0.01 with 3beta-OH-tibolone (n = 3). In conclusion, SULT2A1 is the major endogenous enzyme responsible for sulfation of the tibolone metabolites in human postmenopausal tissues. The results support the occurrence of pre-receptor enzymatic regulation of hydroxytibolone metabolites and prompt further investigation of the tissue-selective regulation of tibolone effects. PMID- 16360723 TI - Duvernoy's gland secretion of Philodryas olfersii and Philodryas patagoniensis (Colubridae): neutralization of local and systemic effects by commercial bothropic antivenom (Bothrops genus). AB - Colubrids involved in human envenomation in Brazil are mainly from the genera Helicops, Oxyrhopus, Thamnodynastes and Philodryas. There is a relatively large number of clinical descriptions involving the Xenodontinae snakes, Philodryas olfersii and Philodryas patagoniensis, in human accidents. The most common manifestations of envenomation are local pain, swelling, erythema and ecchymosis and regional lymphadenopathy with normal coagulation. The aims of this study were to characterize the biochemical and biological properties of P. olfersii and P. patagoniensis venoms, and to investigate their immunological cross-reactivities by using both specific antisera and anti-Bothrops sp serum used for human serum therapy in Brazil, in neutralizing the lethal and hemorrhagic effects of these venoms. We show here that P. olfersii e P. patagoniensis venoms present proteolytic and haemorrhagic activities but are devoid of phospholipase A2 activity. Haemorrhage and lethality induced by P. olfersii and P. patagoniensis are associated with metal-dependent proteinases, since EDTA could block these toxic activities. P. olfersii and P. patagoniensis venoms were immunogenic and the antisera produced were able to recognize several bands in P. olfersii, P. patagoniensis venoms in Bothrops jararaca venom. PMID- 16360724 TI - Proteases from Lonomia obliqua venomous secretions: comparison of procoagulant, fibrin(ogen)olytic and amidolytic activities. AB - The hemorrhagic syndrome caused by Lonomia obliqua caterpillars is an increasing problem in Southern Brazil. The clinical profile is characterized by both hemorrhagic and pro-coagulant symptoms, constituting a paradoxical action of the venom. The effects upon blood coagulation and fibrin(ogen)olysis have been shown to result from the combined action of several active principles found mostly in the bristle extract. The present study reports quali-quantitative differences among L. obliqua secretions: Cryosecretion, hemolymph, bristle extract and tegument extract. Cryosecretion and hemolymph displayed strong amidolytic activity upon several substrates, presented moderated procoagulant activity and high fibrinogen degrading ability. Bristle and tegument extracts presented low amidolytic activity, but bristle extract showed the most potent procoagulant activity and both extracts presented low fibrinogen degrading ability. The differential involvement of these secretions during the accidents with L. obliqua can elucidate the different symptoms presented after envenomation. PMID- 16360725 TI - Identification of wastewater sludge characteristics to predict critical flux for membrane bioreactor processes. AB - The effects of sludge characteristics on critical flux were examined using a submerged membrane bioreactor pilot plant operated under different process conditions to treat municipal wastewater. The sludge in the membrane tank was characterized by measuring colloidal particle concentration, extracellular polymeric substances (EPS), mixed liquor suspended solids (MLSS), temperature, time to filter (TTF) and diluted sludge volume index (DSVI). The colloidal particle concentration was represented by the colloidal total organic carbon (TOC), which is the TOC difference between the filtrate passing through a 1.5 microm pore size filter and the permeate collected from pilot ultrafiltration membrane modules with a pore size of 0.04 microm. The results showed that the critical flux measured by the stepwise flux method was almost solely related to the colloidal TOC despite different sludges tested. In contrast, MLSS was shown to have little impact on the critical flux within the range examined. Neither TTF nor DSVI could be used to reliably predict the critical flux. Furthermore, colloidal TOC can be attributed to soluble EPS, but not bound EPS. Therefore, it is suggested that colloidal TOC be used as a new filterability index for MBR processes in wastewater treatment. PMID- 16360726 TI - A bench-scale aeration study using batch reactors on swine manure stabilization to control odour in post treatment storage. AB - A bench-scale study on swine manure stabilization for odour control was conducted using batch aeration reactors. In trial 1, two aeration lengths, i.e., 0.5 and 4.0 day, were used under uncontrolled ambient temperature that increased gradually over the experimental period. While in trial 2, a 16.0-day aeration scheme was employed under constant 17 degrees C. An airflow rate of 1.2L/s/m(3) was used for both trials to aerate batch reactors containing finishing pig manure with initial total solids (TS) levels ranging from 0.5 to 4.0%. Manure stabilization during the 90-day post-treatment storage was evaluated by the changes in organic materials, nitrogen and volatile fatty acids (VFA). The odour generation potential in the treated manure was determined by the changes in VFA. Up to 827 mL of liquid was lost due to aeration related foaming. The reductions in total volatile solids (TVS), 5-day biochemical oxygen demand (BOD(5)), total Kjeldahl nitrogen (TKN) and VFA during storage were improved when aeration length increased. Low solids levels offered a more advantageous circumstance for manure stabilization and odour control. Biodegradation of organic matter, removal of nitrogen, and breakdown of VFA would increase with increasing ambient temperature. VFA removals in manure under 16.0-day aeration were higher than those under 0.5- and 4.0-day aeration; however, VFA regeneration started to exceed its consumption on day 20 (4 days after the aeration treatment). BOD(5) was the best estimate of VFA concentration in the aerated manure during storage. The 4.0-day aeration scheme was sufficient to stabilize manure to effectively assuage odour generation potential during the 90-day storage under increasing ambient temperature conditions. PMID- 16360728 TI - Essay: Prosthetics for athletes. PMID- 16360729 TI - Profile: Marlon Shirley--paralympic star. PMID- 16360730 TI - Case report: Marathon runner with severe autonomic failure. PMID- 16360731 TI - Profile: Roger Bannister. PMID- 16360732 TI - Essay: The makings of the perfect athlete. PMID- 16360733 TI - Feature: Gene doping. PMID- 16360734 TI - Profile: Richard W Pound, QC--chairman of WADA. PMID- 16360735 TI - Viewpoint: Legalisation of performance-enhancing drugs. PMID- 16360736 TI - Case report: Consequences of ephedra use in an athlete. PMID- 16360737 TI - Profile: Lewis Gordon Pugh--polar swimmer. PMID- 16360738 TI - Essay: Biomechanical support in sport. PMID- 16360739 TI - Feature: Can nutrition improve athletic performance? PMID- 16360740 TI - Essay: Role of the coach in the coach-athlete relationship. PMID- 16360741 TI - Essay: HIV/AIDS prevention and peace through sport. PMID- 16360742 TI - Essay: Aggression in sport. PMID- 16360743 TI - Essay: Risk taking in sport. PMID- 16360744 TI - Profile: Klaus Mees--a hungry man. PMID- 16360745 TI - Essay: Depression in sport. PMID- 16360746 TI - Personal account: A woman tried and tested. PMID- 16360747 TI - Essay: Suspect sex. PMID- 16360748 TI - Essay: Gender verification of female athletes. PMID- 16360749 TI - Essay: Transsexual athletes--when is competition fair? PMID- 16360750 TI - Essay: Physiology of the child athlete. PMID- 16360751 TI - Essay: Injury prevention in young people--time to accept responsibility. PMID- 16360752 TI - Essay: Sudden death in young athletes. PMID- 16360753 TI - Essay: The female athlete triad. PMID- 16360754 TI - Essay: Rehabilitation through sport--pilot project with amputees in Angola. PMID- 16360755 TI - Essay: Fit for life--a geriatrician's perspective on ageing well. PMID- 16360757 TI - Essay: Sport and exercise medicine--a personal perspective. PMID- 16360758 TI - Essay: Does training adversely affect long-term health? PMID- 16360759 TI - Essay: Long-term complications of exercise. PMID- 16360760 TI - Essay: Hunter-gatherer to sedentary lifestyle. PMID- 16360761 TI - Feature: Olympian pins and needles. PMID- 16360763 TI - Personal account: Sports injury of a guide-dog. PMID- 16360764 TI - Profile: Alex Galvez--transitions of fortune. PMID- 16360765 TI - Essay: Manufacturing categories--the case of disabled athletes. PMID- 16360766 TI - Evolution of anthropogenic aerosols in the coastal town of Salina Cruz, Mexico: part I particle dynamics and land-sea interactions. AB - Measurements of aerosol particles in a coastal city in southeast Mexico show that the concentrations and optical properties are strongly linked to land and sea breezes. Maximum concentrations of condensation nuclei (CN), black carbon (BC) and particle bound polycyclic aromatic hydrocarbons (PPAH) occur during land breeze periods and decrease with the sea breeze. The concentrations of particles in air from the ocean, however, remain significantly above background, maritime values as a result of the recirculation of anthropogenic emissions. The mass size distribution is dominated by particles larger than 5 microm when wind speeds exceed 4 m s(-1); otherwise, the uptake of water vapor onto unactivated particles is the process that dominates the growth of particles. Precipitation removes particles larger than 5 microm but CN, BC and PPAH concentrations are minimally affected. PMID- 16360768 TI - Tackling poverty in tuberculosis control. PMID- 16360767 TI - Children's residential exposure to chlorpyrifos: application of CPPAES field measurements of chlorpyrifos and TCPy within MENTOR/SHEDS-Pesticides model. AB - The comprehensive individual field-measurements on non-dietary exposure collected in the Children's-Post-Pesticide-Application-Exposure-Study (CPPAES) were used within MENTOR/SHEDS-Pesticides, a physically based stochastic human exposure and dose model. In this application, however, the model was run deterministically. The MENTOR/SHEDS-Pesticides employed the CPPAES as input variables to simulate the exposure and the dose profiles for seven children over a 2-week post application period following a routine residential and professional indoor crack and-crevice chlorpyrifos application. The input variables were obtained from a personal activity diary, microenvironmental measurements and personal biomonitoring data obtained from CPPAES samples collected from the individual children and in their homes. Simulation results were compared with CPPAES field measured values obtained from the children's homes to assess the utility of the different microenvironmental data collected in CPPAES, i.e. indicator toys and wipe samplers to estimate aggregate exposures that can be result from one or more exposure pathways and routes. The final analyses of the database involved comparisons of the actual data obtained from the individual biomarker samples of a urinary metabolite of chlorpyrifos (TCPy) and the values predicted by MENTOR/SHEDS-Pesticides using the CPPAES-derived variables. Because duplicate diet samples were not part of the CPPAES study design, SHEDs-Pesticides simulated dose profiles did not account for the dietary route. The research provided more confidence in the types of data that can be used in the inhalation and dermal contact modules of MENTOR/SHEDS-Pesticides to predict the pesticide dose received by a child. It was determined that we still need additional understanding about: (1) the types of activities and durations of activities that result in non dietary ingestion of pesticides and (2) the influence of dietary exposures on the levels of TCPy found in the urine. PMID- 16360769 TI - Marketing food to children. PMID- 16360770 TI - British science deserves appropriate scrutiny. PMID- 16360771 TI - The pre-ictal state in focal epilepsy. PMID- 16360772 TI - Psychosocial aid to children after the Dec 26 tsunami. PMID- 16360773 TI - Bone densitometry: an update. PMID- 16360774 TI - Reducing mercury and responding to the global gold rush. PMID- 16360775 TI - Preparing for pandemic influenza. PMID- 16360776 TI - David Barnardo. Interview. PMID- 16360777 TI - Nutrition. PMID- 16360778 TI - Are the clinical effects of homoeopathy placebo effects? PMID- 16360779 TI - Are the clinical effects of homoeopathy placebo effects? PMID- 16360780 TI - Are the clinical effects of homoeopathy placebo effects? PMID- 16360782 TI - Are the clinical effects of homoeopathy placebo effects? PMID- 16360785 TI - Influenza vaccination in elderly people. PMID- 16360786 TI - Effects of radiotherapy and of differences in the extent of surgery for early breast cancer on local recurrence and 15-year survival: an overview of the randomised trials. AB - BACKGROUND: In early breast cancer, variations in local treatment that substantially affect the risk of locoregional recurrence could also affect long term breast cancer mortality. To examine this relationship, collaborative meta analyses were undertaken, based on individual patient data, of the relevant randomised trials that began by 1995. METHODS: Information was available on 42,000 women in 78 randomised treatment comparisons (radiotherapy vs no radiotherapy, 23,500; more vs less surgery, 9300; more surgery vs radiotherapy, 9300). 24 types of local treatment comparison were identified. To help relate the effect on local (ie, locoregional) recurrence to that on breast cancer mortality, these were grouped according to whether or not the 5-year local recurrence risk exceeded 10% (<10%, 17,000 women; >10%, 25,000 women). FINDINGS: About three quarters of the eventual local recurrence risk occurred during the first 5 years. In the comparisons that involved little (<10%) difference in 5-year local recurrence risk there was little difference in 15-year breast cancer mortality. Among the 25,000 women in the comparisons that involved substantial (>10%) differences, however, 5-year local recurrence risks were 7% active versus 26% control (absolute reduction 19%), and 15-year breast cancer mortality risks were 44.6% versus 49.5% (absolute reduction 5.0%, SE 0.8, 2p<0.00001). These 25,000 women included 7300 with breast-conserving surgery (BCS) in trials of radiotherapy (generally just to the conserved breast), with 5-year local recurrence risks (mainly in the conserved breast, as most had axillary clearance and node-negative disease) 7% versus 26% (reduction 19%), and 15-year breast cancer mortality risks 30.5% versus 35.9% (reduction 5.4%, SE 1.7, 2p=0.0002; overall mortality reduction 5.3%, SE 1.8, 2p=0.005). They also included 8500 with mastectomy, axillary clearance, and node-positive disease in trials of radiotherapy (generally to the chest wall and regional lymph nodes), with similar absolute gains from radiotherapy; 5-year local recurrence risks (mainly at these sites) 6% versus 23% (reduction 17%), and 15-year breast cancer mortality risks 54.7% versus 60.1% (reduction 5.4%, SE 1.3, 2p=0.0002; overall mortality reduction 4.4%, SE 1.2, 2p=0.0009). Radiotherapy produced similar proportional reductions in local recurrence in all women (irrespective of age or tumour characteristics) and in all major trials of radiotherapy versus not (recent or older; with or without systemic therapy), so large absolute reductions in local recurrence were seen only if the control risk was large. To help assess the life threatening side-effects of radiotherapy, the trials of radiotherapy versus not were combined with those of radiotherapy versus more surgery. There was, at least with some of the older radiotherapy regimens, a significant excess incidence of contralateral breast cancer (rate ratio 1.18, SE 0.06, 2p=0.002) and a significant excess of non-breast-cancer mortality in irradiated women (rate ratio 1.12, SE 0.04, 2p=0.001). Both were slight during the first 5 years, but continued after year 15. The excess mortality was mainly from heart disease (rate ratio 1.27, SE 0.07, 2p=0.0001) and lung cancer (rate ratio 1.78, SE 0.22, 2p=0.0004). INTERPRETATION: In these trials, avoidance of a local recurrence in the conserved breast after BCS and avoidance of a local recurrence elsewhere (eg, the chest wall or regional nodes) after mastectomy were of comparable relevance to 15-year breast cancer mortality. Differences in local treatment that substantially affect local recurrence rates would, in the hypothetical absence of any other causes of death, avoid about one breast cancer death over the next 15 years for every four local recurrences avoided, and should reduce 15-year overall mortality. PMID- 16360787 TI - Sudden infant death syndrome and complications in other pregnancies. AB - BACKGROUND: The likelihood of recurrence of sudden infant death syndrome (SIDS) is an issue of biological, clinical, and legal interest. Obstetric complications are associated with an increased risk of SIDS and are likely to recur in subsequent pregnancies. We postulated that women whose infants died from SIDS would be more likely to have had obstetric complications in their other pregnancies. METHODS: We linked national UK databases of maternity-hospital discharges, perinatal deaths, and death certifications. We studied 258 096 women who had consecutive births in Scotland between 1985 and 2001. FINDINGS: Women who had an infant who died from SIDS were at increased risk in their next pregnancy of delivering an infant small for gestational age (odds ratio 2.27, 95% CI 1.54 3.34, p<0.0001) and of preterm birth (2.53, 1.82-3.53, p<0.0001). The risk of SIDS was higher for the children of women whose previous infant had been small for gestational age (1.87, 1.19-2.94, p=0.007) or preterm (1.93, 1.24-3.00, p=0.004). Multivariate analysis showed that all associations were explained by common maternal risk factors for SIDS and obstetric complications and by the likelihood of recurrence of fetal growth restriction and preterm birth. INTERPRETATION: Women whose infants die from SIDS are more likely to have complications in their other pregnancies. Recurrence of pregnancy complications predisposing to SIDS could partly explain why some women have recurrent SIDS. PMID- 16360788 TI - Global prevalence of dementia: a Delphi consensus study. AB - BACKGROUND: 100 years after the first description, Alzheimer's disease is one of the most disabling and burdensome health conditions worldwide. We used the Delphi consensus method to determine dementia prevalence for each world region. METHODS: 12 international experts were provided with a systematic review of published studies on dementia and were asked to provide prevalence estimates for every WHO world region, for men and women combined, in 5-year age bands from 60 to 84 years, and for those aged 85 years and older. UN population estimates and projections were used to estimate numbers of people with dementia in 2001, 2020, and 2040. We estimated incidence rates from prevalence, remission, and mortality. FINDINGS: Evidence from well-planned, representative epidemiological surveys is scarce in many regions. We estimate that 24.3 million people have dementia today, with 4.6 million new cases of dementia every year (one new case every 7 seconds). The number of people affected will double every 20 years to 81.1 million by 2040. Most people with dementia live in developing countries (60% in 2001, rising to 71% by 2040). Rates of increase are not uniform; numbers in developed countries are forecast to increase by 100% between 2001 and 2040, but by more than 300% in India, China, and their south Asian and western Pacific neighbours. INTERPRETATION: We believe that the detailed estimates in this paper constitute the best currently available basis for policymaking, planning, and allocation of health and welfare resources. PMID- 16360789 TI - Mitochondrial DNA and survival after sepsis: a prospective study. AB - BACKGROUND: Human genome evolution has been shaped by infectious disease. Although most genetic studies have focused on the immune system, recovery after sepsis is directly related to physiological reserve that is critically dependent on mitochondrial function. We investigated whether haplogroup H, the most common type of mitochondrial DNA (mtDNA) in Europe, contributes to the subtle genetic variation in survival after sepsis. METHODS: In a prospective study, we included 150 individuals who were sequentially admitted to the intensive care unit in a hospital in Newcastle upon Tyne, UK. After clinical data were obtained, patients underwent mtDNA haplotyping by analysis with PCR and restriction fragment length polymorphism. As endpoints, we used death during the 6-month period or survival at 6 months. FINDINGS: Follow-up was complete for all study participants, although the haplotype of two patients could not be reliably determined. On admission to the intensive care unit, the frequency of mtDNA haplogroup H in study patients did not differ between study patients admitted with severe sepsis and 542 age-matched controls from the northeast of England. MtDNA haplogroup H was a strong independent predictor of outcome during severe sepsis, conferring a 2.12-fold (95% CI 1.02-4.43) increased chance of survival at 180 days compared with individuals without the haplogroup H. INTERPRETATION: Although haplogroup H is the most recent addition to the group of European mtDNA, paradoxically it is also the most common. Increased survival after sepsis provides one explanation for this observation. MtDNA haplotyping offers a new means of risk stratification of patients with severe infections, which suggests new avenues for therapeutic intervention. PMID- 16360790 TI - A spinal foreign body. PMID- 16360791 TI - Sex-work harm reduction. AB - Sex work is an extremely dangerous profession. The use of harm-reduction principles can help to safeguard sex workers' lives in the same way that drug users have benefited from drug-use harm reduction. Sex workers are exposed to serious harms: drug use, disease, violence, discrimination, debt, criminalisation, and exploitation (child prostitution, trafficking for sex work, and exploitation of migrants). Successful and promising harm-reduction strategies are available: education, empowerment, prevention, care, occupational health and safety, decriminalisation of sex workers, and human-rights-based approaches. Successful interventions include peer education, training in condom-negotiating skills, safety tips for street-based sex workers, male and female condoms, the prevention-care synergy, occupational health and safety guidelines for brothels, self-help organisations, and community-based child protection networks. Straightforward and achievable steps are available to improve the day-to-day lives of sex workers while they continue to work. Conceptualising and debating sex-work harm reduction as a new paradigm can hasten this process. PMID- 16360792 TI - The road. PMID- 16360793 TI - Prosper Meniere. PMID- 16360794 TI - Giving credit where credit is due: John Hunter and the discovery of erythrocyte sedimentation rate. PMID- 16360795 TI - Potential of cooperation between human and animal health to strengthen health systems. PMID- 16360796 TI - Lead poisoning--a differential diagnosis for abdominal pain. PMID- 16360797 TI - Genetic background of type I protein C deficiency in Finland. AB - In contrast to other populations the usually rare type II form of protein C deficiency is as common in Finland as type I deficiency. We recently reported that a single mutation explained virtually all cases of type II protein C deficiency in Finland, indicating strong founder effect. We now investigated in the same population the genetic background of type I protein C deficiency. Thirty eight apparently unrelated families were studied. They represent the vast majority of all families with type I deficiency in Finland. A genetic defect was identified in 23 (61%) families who carried 13 different mutations. Only three of the 13 mutations have been reported in other populations. Unlike in type II deficiency, considerable heterogeneity in mutations was found in type I deficiency. Our results indicate interesting differences in mutational histories of these two different forms of protein C deficiency in Finland. PMID- 16360798 TI - Development of polyphenol oxidase activity in the micropylar endosperm of tomato seeds. AB - Polyphenol oxidase (PPO) activity increased markedly in the micropylar region of the endosperm of tomato (Lycopersicon esculentum) seeds after radicle protrusion. Tissue-printing analyses demonstrated that the majority of the activity is localized in the micropylar area. The increase in the activity was due to the increase in the amounts of enzyme. Within the micropylar endosperm region, two PPO isozymes were immunologically detected whose apparent molecular masses were estimated to be approximately 58 and 59 kDa, respectively, by SDS-PAGE. Although PPO activity also developed in the lateral portion of the endosperm, the level of this activity was much lower as compared with that in the micropylar region. Furthermore, the isozyme pattern in the lateral portion differed from that in the micropylar portion. The 58 kDa isozyme that was detected in the latter portion was absent, and only 59 kDa PPO was detectable in the former one. When the endosperm tissues were wounded, an enhancement of the enzyme activity was observed in the wounded region. The wound-induced development of the enzyme activity was associated with the induction of 58 kDa isozyme. PMID- 16360799 TI - Effect of salinity and different nitrogen sources on the activity of antioxidant enzymes and indole alkaloid content in Catharanthus roseus seedlings. AB - The activities of antioxidant enzymes viz. glutathione reductase, GR; superoxide dismutase, SOD; peroxidase, POD; catalase, CAT and glutathione-S-transferase, GST and alkaloid accumulation were investigated in leaf pairs (apical, middle, basal) and in roots of Catharanthus roseus seedlings under the conditions of different nitrogen sources (20 mM KNO(3) and 2 mM NH(4)Cl) and salinity, in the absence (non-saline control) and in the presence of 100 mM NaCl in the nutrient solution. Salinity caused a reduction in plant biomass. The biomass production of ammonium fed plants was lower than that of nitrate-fed plants. The antioxidant enzymes exhibited higher activity in saline-treated plants. Changes in antioxidant enzyme activity caused by different nitrogen sources differed in all leaf pairs, as well as in roots of C. roseus. Ammonium-fed plants showed higher CAT, GR and GST activity in leaf pairs as well as in roots, while POD and SOD activity were higher in nitrate-fed plants. Higher peroxidase activity concomitant with the increased accumulation of alkaloid was found in all leaf pairs, as well as in roots of C. roseus of NO(3)(-) fed plants as compared to NH(4)(+) fed plants. PMID- 16360800 TI - Immunolocalization of a class III chitinase in two muskmelon cultivars reacting differently to Fusarium oxysporum f. sp. melonis. AB - Fusarium oxysporum f. sp. melonis is a highly specialized fungus that attacks the root system of melon (Cucumis melo L.). In this work the presence of a class III chitinase was examined by immunological techniques in the root and stem base of a susceptible (cv. Galia) and a resistant (cv. Bredor) melon during the infection process. By immunolocalization it was not possible to detect the constitutive presence of class III chitinase in any of the cultivars. However, the immunolabelling appeared in the root tissues of both cultivars as a consequence of wounding and of infection by F. oxysporum f. sp. melonis. Distinct patterns of chitinase detection were observed in the roots of the two cultivars as the infection progressed. Furthermore, by western blotting distinct class III chitinase isoforms were detected, which responded differently to the F. oxysporum f. sp. melonis infection. Our results strongly indicate that a relationship exists between class III chitinase and melon resistance to Fusarium infection, and that the resistance is associated with certain isoforms of this enzyme. PMID- 16360801 TI - A possible role of sphingolipids in the aluminium resistance of yeast and maize. AB - The plasma membrane is most likely the major target for sensing of aluminium (Al), leading to inhibition of plant root-growth. As a result of high external Al, alterations in plasma membrane composition may be expected in order to maintain its properties. As sphingolipids are characteristic components of this membrane, their involvement in membrane adjustment to increased Al concentrations was investigated. Heterologous expression of a stereounselective long-chain base (LCB) (8E/Z)-desaturase from Arabidopsis thaliana, Brassica napus and Helianthus annuus in Saccharomyces cerevisiae improved the Al resistance of the transgenic yeast cells. This encouraged us to investigate whether Al affects the LCB composition, and whether genetic engineering of the LCB profile modifies the Al resistance of the Al-sensitive plant species maize (Zea mays, L.). Constitutive expression of the LCB (8E/Z)-desaturase from Arabidopsis thaliana in maize roots led to an 8- to 10-fold increase in (8E)-4-hydroxysphing-8-enine in total roots. Less marked but similar changes were observed in 3 mm root apices. Al treatment of the Al-sensitive maize cv Lixis resulted in a significant increase in the proportion of (8Z)-LCB and in the content of total LCBs in root tips, which was not observed in the Al-resistant cv ATP-Y. When root tips of transgenic plants were exposed to Al, only minor changes of both (8Z)- and (8E)-unsaturated LCBs as well as of the total LCB were observed. Al treatment of the wild type parental line H99 decreased the (8Z)-unsaturated LCBs and the total LCB content. Based on Al-induced callose production, a marker for Al sensitivity, the parental line H99 was as Al-resistant as cv ATP-Y, whereas the transgenic line became as sensitive as cv Lixis. Taken together, these data suggest that, in particular, the loss of the ability to down-regulate the proportion of (8Z)-unsaturated LCBs may be related to increased Al sensitivity. PMID- 16360802 TI - Growth and cell wall properties of two wheat cultivars differing in their sensitivity to aluminum stress. AB - The present study was conducted to investigate the cell wall properties in two wheat (Triticum aestivum L.) cultivars differing in their sensitivity to Al stress. Seedlings of Al-resistant, Inia66 and Al-sensitive, Kalyansona cultivars were grown in complete nutrient solutions for 4 days and then subjected to treatment solutions containing Al (0, 50 microM) in a 0.5 mM CaCl(2) solution at pH 4.5 for 24 h. Root elongation was inhibited greatly by the Al treatment in the Al-sensitive cultivar compared to the Al-resistant cultivar. The Al-resistant cultivar accumulated less amount of Al in the root apex than in the Al-sensitive cultivar. The contents of pectin and hemicellulose in roots were increased with Al stress, and this increase was more conspicuous in the Al-sensitive cultivar. The molecular mass of hemicellulosic polysaccharides was increased by the Al treatment in the Al-sensitive cultivar. The increase in the content of hemicellulose was attributed to increase in the contents of glucose, arabinose and xylose in neutral sugars. Aluminum treatment increased the contents of ferulic acid and p-coumaric acid especially in the Al-sensitive cultivar by increasing the activity of phenylalanine ammonia lyase (PAL, EC 4.3.1.5). Aluminum treatment markedly decreased the beta-glucanase activity in the Al sensitive cultivar, but did not exert any effect in the Al-resistant cultivar. These results suggest that the modulation of the activity of beta-glucanase with Al stress may be involved in part in the alteration of the molecular mass of hemicellulosic polysaccharides in the Al-sensitive cultivar. The increase in the molecular mass of hemicellulosic polysaccharides and ferulic acid synthesis in the Al-sensitive cultivar with Al stress may induce the mechanical rigidity of the cell wall and inhibit the elongation of wheat roots. PMID- 16360803 TI - Season effects on leaf nitrogen partitioning and photosynthetic water use efficiency in mango. AB - The key parameters of photosynthetic capacity (maximum carboxylation rate (V(cmax)), electron transport capacity (J(max)) and dark respiration rate (R(d))) and the slope (m) of the stomatal conductance model of Ball et al. [Progress in photosynthetic research, Martinus Nijhoff, Dordrecht, 1987] were measured for a whole growing season in fully expanded leaves of 12-year-old mango trees cv. Cogshall in La Reunion island. Leaf nitrogen partitioning into carboxylation (P(c)) and bioenergetic (P(b)) pools were computed according to the model of Niinemets and Tenhunen [Plant Cell Environ 1997;20: 845-66]. V(cmax), J(max), R(d), P(c) and P(b) remained relatively stable over the whole study period, with the exception of the period of linear fruit growth when J(max), R(d) and P(b) were slightly lower, and leaf non-structural carbohydrate content higher. During the pre-floral and floral periods, m decreased by more than 50%, indicating an increase in photosynthetic water use efficiency and m increased again during the period of linear fruit growth. Our results show that, in tropical orchard conditions characterized by mild seasonal climatic changes and non-limiting water supply, leaf nitrogen partitioning is rather stable. Our results also advocate for more studies on the effect of phenology on m and photosynthetic water use efficiency, which is of paramount importance for building coupled biochemical models of photosynthetic carbon assimilation. PMID- 16360804 TI - Molecular cloning and expression analysis of the replacement histone H3 gene of Italian ryegrass (Lolium multiflorum). AB - The replacement histone H3 gene and its 5'-flanking sequence were isolated from Italian ryegrass by polymerase chain reaction and inverse polymerase chain reaction, respectively. Expression analysis showed that this gene is constitutively expressed in the entire plant. The expression level in leaves was found to be significantly low when compared with that in other tissues. However, the gene expression level in leaves was increased by the treatment with abscisic acid and abiotic stresses such as cold, heat and high-salinity (NaCl). The motif search of the 5'-flanking sequence of the replacement histone H3 gene revealed the presence of several potential cis-acting elements that could respond to the above-mentioned abiotic stresses. In addition to defence-related elements, we also found type I and II-/III-like elements, which are highly conserved motifs in the 5'-regulatory sequence of plant histone genes that are expressed specifically during the S-phase. Experiments using transgenic Italian ryegrass plants proved that the isolated 5'-flanking sequence of the replacement histone H3 gene, which was fused to a beta-glucuronidase reporter gene, was fully functional for inducing gene expression under various abiotic stress conditions. PMID- 16360805 TI - Comparative study of rice and Arabidopsis actin-depolymerizing factors gene families. AB - Actin-depolymerizing factors (ADF) is one of the small actin-binding proteins that regulate actin dynamics in cells. Analysis of the complete rice and Arabidopsis protein sequences revealed 12 ADF proteins, respectively. A further study on the similarities and differences between ADF throughout rice and Arabidopsis genome was carried out at the level of genomic organization and protein structure. The strict conservation of essential structural features suggested that the mode of action and physiological function of these proteins, as well as the expression pattern of their coding genes, might be very similar. The ADF proteins were divided into four groups based on the phylogenetic relationships of the amino acid sequences, and was comparable with previous studies. PMID- 16360806 TI - Nitrogen compounds in the apoplastic sap of sugarcane stem: some implications in the association with endophytes. AB - Several nitrogen compounds were identified and quantified in the apoplastic and symplastic sap of sugarcane stems. The sap of stems was composed mainly of soluble sugars, which constituted 95% of the total organic compounds detected. Sap also contained nitrogen compounds, with amino acids (50-70% of N) and proteins (20-30% of N), being the main nitrogenous substances, as well as inorganic forms as ammonium, nitrite and nitrate, in low concentrations (<20% of N). Serine, proline, alanine and aspartic acid together represented around 60% of the amino acids of the sap of both field grown and high nitrogen fertilized plants, and non-nitrogen fertilized plants inoculated with Gluconacetobacter diazotrophicus. The total amino acid content of apoplastic sap was six to nine times lower in non-nitrogen fertilized plants than in fertilized ones. The possible roles of these substances to regulate endophytic associations with sugarcane are also discussed. PMID- 16360807 TI - Flower fertilization and fruit development prompt changes in free polyamines and ethylene in damson plum (Prunus insititia L.). AB - The flower opening of damson plum (Prunus insititia L.) was accompanied by an increase in the content of free-polyamines (PA) in the sepals, petals and sex organs, the ovary being most active in accumulating spermine (Spm). The fertilization process and senescence brought on a decline in ovarian Spm, but stimulated putrescine (Put) and spermidine (Spd) content in the sepals. The endocarp of this climacteric fruit produced only ethylene at the end of the S1 phase and throughout S2, in which there was a great richness in ACC and MACC. The greatest amounts of ACC and MACC were observed in the ripening mesocarp and epicarp. The contribution of the endocarp and epicarp to the total ACC in the developing fruit was very similar. During flowering and S1 and S2 phases, Spd was the most abundant PA; in contrast, during S3 and S4 Put was most abundant. The mesocarp contributed the most to the total content in PA throughout the fruit development. The control of SAM distribution towards ethylene and/or PA appears to differ during the development of the endocarp, as the only peak of free-Put (detected in S2) coincided with the highest ACC accumulation and ethylene production. On the contrary, in S3 it is probable that SAM was transformed preferentially into PA, given that free-Spd and Spm, hardly detectable in S1 and S2, peaked in this phase in which there was no gas production. PMID- 16360808 TI - Plant regeneration through multiple adventitious shoot differentiation from callus cultures of slash pine (Pinus elliottii). AB - A plant regeneration system through multiple adventitious shoot differentiation from callus cultures has been established in slash pine (Pinus elliottii). Influences of seven different basal media on callus induction, adventitious shoot formation, and rooting were investigated. Among the different basal media, B5, SH, and TE proved to be suitable for callus induction and plantlet regeneration. Multiple adventitious shoot formation was obtained from callus cultures of slash pine on B5, SH, and TE media containing indole-3-butyric acid, N6-benzyladenine, and thidiazuron. Scanning electron microscopy demonstrated the early development of adventitious shoots derived from callus cultures. These results indicate that an efficient plant regeneration protocol for micropropagation of slash pine had been established. This protocol could be most useful for future studies on genetic transformation of slash pine. PMID- 16360809 TI - Sugarbeet sucrose synthase genes differ in organ-specific and developmental expression. AB - A full-length sucrose synthase (SBSS2) cDNA clone was isolated from sugarbeet. Comparison of its composition and organ-specific and developmental expression with a previously isolated sugarbeet sucrose synthase gene (SBSS1) revealed distinct differences between the two genes. The two genes share 80% similarity in deduced amino acid sequence but belong to different sucrose synthase subclasses based on phylogenic analysis. Both sucrose synthases were highly expressed in roots, and had low levels of expression in leaf tissue. Transcript abundance of SBSS2, relative to SBSS1, was greater in young vegetative and floral tissues, and reduced in mature vegetative tissues. The organ-specific and developmental expression of SBSS1 and SBSS2 proteins was similar to SBSS1 and SBSS2 transcript levels, although developmental changes in protein abundance lagged transcriptional changes by many weeks. The similarities and differences in transcript and protein abundance suggest that both transcriptional and post transcriptional regulatory mechanisms are likely to contribute to sucrose synthase expression in sugarbeet. PMID- 16360810 TI - Nod Bj-V (C18:1, MeFuc) production by Bradyrhizobium japonicum (USDA110, 532C) at suboptimal growth temperatures. AB - Nod factors (Lipo-chitooligosaccharides, or LCOs) act as bacteria-to-plant signal molecules that modulate early events of the Bradyrhizobium-soybean symbiosis. It is known that low root zone temperature inhibits the early stages of this symbiosis; however, the effect of low soil temperature on bacteria-to-plant signaling is largely uninvestigated. We evaluated the effect of low growth temperatures on the production kinetics of Nod factor (LCO) by B. japonicum. Two strains of B. japonicum, 532C and USDA110, were tested for ability to synthesize Nod Bj-V (C(18:1), MeFuc) at three growth temperatures (15, 17 and 28 degrees C). The greatest amounts of the major Nod factor, Nod Bj-V (C(18:1), MeFuc), were produced at 28 degrees C for both strains. At 17 and 15 degrees C, the Nod factor production efficiency, per cell, of B. japonicum 532C and USDA110 was markedly decreased with the lowest Nod factor concentration per cell occurring at 15 degrees C. Strain 532C was more efficient at Nod factor production per cell than strain USDA 110 at all growth temperatures. The biological activity of the extracted Nod factor was unaffected by culture temperature. This study constitutes the first demonstration of reduced Nod factor production efficiency (per cell production) under reduced temperatures, suggesting another way that lower temperatures inhibit establishment of the soybean N(2) fixing symbiosis. PMID- 16360811 TI - Two functionally relevant polymorphisms in the human progesterone receptor gene (+331 G/A and progins) and the predisposition for breast and/or ovarian cancer. AB - OBJECTIVE: Two polymorphisms affecting either expression (+331 G/A) or transcriptional activity (progins) of the progesterone receptor have been described. No clear correlation between either polymorphism and breast or ovarian cancer has been shown. Our objective is to clarify whether the two progesterone receptor polymorphisms modify the risk for breast or ovarian cancer. METHODS: Healthy women and women suffering from either ovarian or breast cancer were enrolled in a case-control-based study to compare the frequencies of women carrying either one, both or none of the two polymorphisms. Patient and control populations resided in the same region of South Germany. PCR-RFLP analysis was used to determine the polymorphic alleles. RESULTS: Women diagnosed with ovarian cancer showed a not significant increased frequency of +331 A carriers and a significantly increased frequency of progins carriers. Both polymorphisms appeared to be associated with a significantly increased risk for the disease in women below 51 years [OR: 4.1 (CI: 1.2-13.9) and 3.2 (CI: 1.1-9.1), respectively]. No association was detected between either of the two polymorphisms and breast cancer. Among ovarian and breast cancer patients, the number of individuals carrying both rare polymorphic alleles was significantly higher compared to healthy women. CONCLUSIONS: Our findings support the hypothesis that low penetrant polymorphisms of progesterone receptor may modify the risk for ovarian cancer. Our data do not allow drawing a clear conclusion on the risk for breast cancer. PMID- 16360812 TI - BRCA1 promoter methylation predicts adverse ovarian cancer prognosis. AB - OBJECTIVE: To compare the clinical outcome of ovarian cancer patients whose tumors contain BRCA1 genes silenced by promoter hypermethylation to patients with germline BRCA1 mutations and to patients with wild-type BRCA genes. METHODS: Ovarian cancers from a hospital-based tumor bank were characterized as having a BRCA1 mutation; or a methylated BRCA1, BRCA1 pseudogene or MLH1 promotor; or a wild-type BRCA gene. Survival of patients with methylated BRCA1 promoters (N = 11) was compared to that of patients with wild-type BRCA genes (N = 30) and BRCA1 mutations (N = 22). A methylator phenotype was defined to include tumors with hypermethylation of BRCA1, hMLH1 and/or dBRCA1 pseudogene promoters (N = 23). RESULTS: All cohorts had comparable clinical factors except for age at diagnosis. Median age of methylated BRCA1 and wild-type BRCA patients was older than BRCA1 mutation carriers (60 and 63 versus 48 years; P = 0.04). The median disease-free interval was significantly shorter for patients with a methylated BRCA1 promoter (9.8 months) than for BRCA1 mutation carriers (39.5 months; P = 0.04). Median overall survival was also significantly shorter for patients with a methylated BRCA1 promoter (35.6 months) than BRCA1 mutation carriers (78.6 months; P = 0.02). The combined methylator phenotype cohort had significantly shorter survival (36.1 months) compared to wild-type BRCA patients (63.3 months; P = 0.02). CONCLUSION: These data suggest that methylation of the BRCA1 promoter is associated with poor patient outcome. BRCA1 may be part of a global panel of methylated genes associated with aggressive disease. PMID- 16360813 TI - Acute effects of nicotine on attention and response inhibition. AB - Smoking is highly prevalent among patients with Attention Deficit Hyperactivity Disorder (ADHD). Previous studies using the reversed continuous performance task (R-CPT) have suggested that nicotine reduces inattention. Since especially adults with ADHD have been claimed to suffer from a core deficit in inhibitory control, this study aimed at determining whether nicotine improves response inhibition in addition to attention. Sixteen healthy regular smokers participated in a pre/post treatment design in which transdermal patches containing 7 and 21 mg nicotine per day were administered in a counterbalanced, double-blind manner. In a second study, patches containing 0 mg (placebo) and 21 mg per day were administered to a different group of regular smokers. For replication purposes, the R-CPT and the profile of mood states (POMS) were administered. Furthermore, a different version of the continuous performance task (CPT-AX) and the stop-signal task, traditionally used to measure response inhibition, were presented. The high dose of nicotine was found to relieve self-reported Depression in Study 1 and Fatigue in Study 2. Performance data indicated acute effects of nicotine on attention related, but not on inhibition-related measures. Especially the comparison with placebo revealed decreases in reaction time and variability of responding. The results imply that patients with ADHD smoke to reduce inattention. PMID- 16360814 TI - Squamous cell carcinoma of the subglottis. AB - STUDY DESIGN: A retrospective study over a 25-year period of patients with laryngeal carcinoma treated by the Department of Otolaryngology at Wake Forest University. METHODS: The boundaries of the subglottis were defined as 5 mm below the free edge of the true vocal folds extending to the inferior border of the cricoid cartilage. All were staged according to American Joint Committee on Cancer: stages I and II were considered early and stages III and IV as late. Patients were grouped by treatment modality of surgery alone, surgery and radiotherapy, radiotherapy alone, and radiotherapy with surgical salvage. RESULTS: Fifteen patients represented 1.4% (15/1098) of laryngeal cancers diagnosed within that period. All patients had squamous cell carcinoma of the subglottis of which 20% (3/15) had early-stage disease (T1-T2) and 80% (12/15) had late-stage disease (T3-T4). Overall 3-year survival was low (25%) and is attributed to a high incidence of advanced-stage disease with a high rate of extralaryngeal spread and/or metastasis, especially to the lungs and paratracheal nodes. CONCLUSIONS: To improve earlier detection by primary care physicians and otolaryngologists and to improve treatment outcome, awareness of subglottic carcinoma and its appropriate evaluation is critical in the patient presenting with hoarseness and/or stridor. Radiation therapy treatment to include the low and upper mediastinal compartments to cover local/regional extralaryngeal involvement is advocated as well as paratracheal lymphatic neck dissection and thyroidectomy in surgically treated lesions. PMID- 16360815 TI - The pattern of lymphatic metastasis of malignant tumors in the periorbital area. AB - BACKGROUND AND OBJECTIVES: The rarity of malignant tumors in the periorbital area makes it difficult to assess the patterns of lymphatic spread and to determine the optimal extent of treatment. The objective of this study was to analyze the patterns of lymphatic metastasis in malignant tumors arising from the periorbital area according to the primary site and to assess the optimal extent of surgical treatment for these tumors. MATERIALS AND METHODS: We retrospectively reviewed the medical records of 8 patients with malignant tumors in the periorbital area from January 1997 to June 2002. The mean duration of follow-up was 27.5 months. Five primary tumors were located in the medial half of the periorbital area, 2 tumors in both medial and lateral halves, and 1 tumor in the lateral half. The pathological diagnoses were 3 squamous cell carcinomas, 2 adenoid cystic carcinomas, 2 sebaceous carcinomas, and 1 basal cell carcinoma. RESULTS: The lymphatic metastasis was observed in 4 of 8 patients who had a relatively large (> or =4 cm) primary tumor or poorly differentiated tumor. Four of 7 tumors involving the medial half of the periorbital area exhibited lymphatic spread to the periparotid lymph node and level II cervical lymph node groups. CONCLUSION: The results of this study suggest that more aggressive management of the parotid area is necessary in malignant tumors arising from the periorbital area, even in the medial half of the periorbital area. PMID- 16360816 TI - The potential impact of palivizumab on pediatric airway reconstruction. AB - PURPOSE: To examine the role of palivizumab, a monoclonal antibody vaccine designed to prevent respiratory syncytial virus (RSV) infection, in the surgical setting, and to explore the feasibility, in terms of medical benefit and cost, of its use in pediatric airway reconstruction patients. MATERIALS AND METHODS: Literature review of MEDLINE database using the following indexing terms ("respiratory syncytial virus," "palivizumab," "laryngotracheal reconstruction," and "cricotracheal reconstruction") and limited to following subcategories (English language, postoperative complication, and morbidity and mortality). All phase IV clinical trials reporting data regarding safety, efficacy, and application of palivizumab in the general pediatric population were selected, as well as all studies from any surgical specialty with data on postoperative outcome complicated by RSV infection. RESULTS: The literature demonstrates significant postoperative morbidity and mortality attributable to RSV infection within several surgical specialties including otolaryngology. Meta-analysis of the data from phase IV clinical trials suggests potential benefit from the perioperative use of palivizumab. The cost of vaccination is the principal limiting factor preventing its more widespread application. CONCLUSION: Otolaryngologists need to be aware of the potential significant morbidity caused by perioperative RSV infection and should consider the prophylactic use of palivizumab in their pediatric airway reconstruction patients in high-risk seasons. A large multicenter study would be required to adequately perform a cost benefit analysis of palivizumab use for this specific indication. PMID- 16360817 TI - The effect of donor-specific transfusion upon rejection in a rat model of laryngeal transplantation. AB - Lifelong immunosuppression carries significant morbidity, and techniques to reduce or eliminate such immunosuppression might expand laryngeal transplantation. This study investigates the ability of donor-specific transfusion to establish tolerance in a rat model of laryngeal transplantation. A total of 289 transplants were performed from Lewis-Brown-Norway donors to Lewis recipients. Donor-specific transfusion was provided as single intravenous injections of donor splenocytes 1 hour before transplantation. Different combinations of in vitro irradiation of donor larynges and postoperative cyclosporine doses provided additional immunomodulation. Allograft rejection was scored histologically at sacrifice 15 or 30 days posttransplant. Multivariate analysis was performed across all groups, and paired analyses compared groups with and without donor-specific transfusion. Donor-specific transfusion did not improve rejection score, although increased cyclosporine dose did (P < .001). Donor splenocyte injection on the day of transplantation does not establish tolerance to laryngeal allografts or permit reduction of other immunosuppression. Other immunomodulatory strategies to establish tolerance in rat laryngeal transplantation require further investigation. PMID- 16360818 TI - Physician hearing loss. AB - PURPOSE: Hearing is an important sense for physicians, making communication and stethoscope use possible, yet not much is known about the impact of hearing loss on professional function. The purpose of this study was to explore hearing related issues affecting physicians. MATERIALS AND METHODS: We administered a hearing test and questionnaire to 107 physicians and medical students. RESULTS: The proportion of physicians reporting trouble with their hearing increased with age, reaching almost 100% in those older than 60 years. Audiometric hearing loss also increased with age. Perceived hearing trouble was significantly associated with audiometric hearing loss, yet 46% of physicians with hearing loss described their hearing as good. Older physicians more frequently reported difficulty communicating with patients, staff, and colleagues owing to hearing problems (P = .007). Reported stethoscope difficulties did not significantly increase with age; there was no association with hearing thresholds. No physician reported use of electronic stethoscopes or hearing aids. Noise exposures were common, yet 51% of respondents never used hearing protection. Younger physicians were less likely to use protection (P = .002). CONCLUSION: Physicians lose hearing with age but may not notice or report the loss. Physician hearing loss is associated with difficulty communicating with patients, staff, and colleagues. Neither age nor hearing level predicts problems with stethoscope use; possible explanations include a training effect or denial. Many physicians, especially younger ones, never use hearing protection around noise. Strategies to recognize and reduce the impact of hearing loss on professional function throughout a physician's career deserve greater attention. PMID- 16360819 TI - External beam radiotherapy for differentiated thyroid cancer. AB - PURPOSE: To evaluate the efficacy of external beam radiation therapy (EBRT) for differentiated thyroid carcinomas. METHODS: Forty-two patients with a locally advanced or recurrent differentiated thyroid carcinoma received high-dose EBRT. Thirty-three patients had local-regional disease and 9 patients also had asymptomatic disease metastases. Twenty patients (48%) had macroscopic local regional disease before EBRT. RESULTS: The 5-year outcomes for the overall population and for the subset of those without distant metastases before EBRT were local-regional control (88% and 89%, respectively), cause-specific survival (80% and 86%, respectively), and overall survival (54% and 60%, respectively). Local-regional control was improved for patients with microscopic residual disease compared with macroscopic disease, those with previously untreated tumors, and for those who received more than 64 Gy. CONCLUSION: EBRT increases the likelihood of local-regional control for patients with locally advanced differentiated thyroid carcinoma. PMID- 16360820 TI - Evaluation of fibula free flap donor site morbidity. AB - OBJECTIVE: To evaluate foot and ankle function in a series of patients undergoing fibula microvascular free tissue transfer. STUDY DESIGN: A nonrandomized, nonblinded analysis was performed using the ankle-hindfoot scale as well as radiographic analysis to determine donor site morbidity. RESULTS: Overall, foot ankle function was deemed to be good in this series of patients. Average ankle hindfoot score was 84.82 of a possible 100 (range, 55-100). Radiographic analysis of donor and contralateral ankles showed no difference in the syndesmotic space or medial clear space. Only 1 patient had increased talar tilt compared with the contralateral side, although this patient had preservation of stability. CONCLUSION: Fibula free flap harvest appears to be associated with acceptable donor site morbidity and preservation of good foot and ankle function in most individuals. PMID- 16360821 TI - Characteristics of sensorineural hearing loss in children with inner ear anomalies. AB - PURPOSE: To determine whether hearing loss in children with inner ear anomalies has some distinctive characteristics when compared to children with hearing loss but without inner ear anomalies. METHODS: Temporal bone computed tomography scans of 69 patients with sensorineural hearing loss were examined for inner ear abnormalities of which 17 were identified. The medical histories of these patients were reviewed for the characteristics of their hearing loss, including initial presentation, natural history, and nature of loss, as well as the family history of hearing loss and risk factors for hearing loss. These were compared to age-matched controls with hearing loss but without inner ear anomalies. RESULTS: Seventeen patients had inner ear anomalies. Records of 14 of these patients were compared to patients without inner ear anomalies. Regarding age of onset, 71.4% of patients with anomalies had onset of their hearing loss at less than 2 years old vs 78.6% without anomalies. Regarding unilateral vs bilateral, 42.9% of patients with anomalies were unilateral vs 28.6% of patients without anomalies. For patients with anomalies, 85.7% were stable and 14.3% were progressive; without anomalies, 71.4% were stable, 21.4% were progressive, and 7.1% were fluctuating. Regarding family history, only 14.3% of patients without anomalies had a positive family history vs 56% of patients with anomalies. CONCLUSIONS: Children with inner ear anomalies and sensorineural hearing loss have an increased incidence of unilateral hearing loss and stable hearing loss as compared to controls with sensorineural hearing loss without inner ear anomalies. In addition, children with inner ear anomalies and sensorineural hearing loss are less likely to have a family history of hearing loss. PMID- 16360822 TI - Tuberculosis in ear, nose, and throat practice: its presentation and diagnosis. AB - PURPOSE: The aim of this study was to increase awareness of the different presentations of head and neck tuberculosis (TB) and to discuss its diagnostic difficulties. MATERIALS AND METHOD: A retrospective analysis of patients who presented to us, at a secondary referral hospital, primarily with TB of head and neck was done from January 1999 to July 2003. RESULTS: A total of 117 patients presented with primary head and neck TB during the study period. Most of these (95%) had cervical lymphadenopathy, 2 patients had laryngeal TB, and there was 1 patient each of TB of cervical spine, oropharynx, ear, and retropharyngeal abscess. Forty-one were males, and 76 were females. Thirty percent of cases had associated lung or other organ TB. Nine percent gave history of previous or subsequent TB. CONCLUSIONS: 1) Diagnosing TB requires a high index of suspicion. 2) Tuberculosis of the cervical lymph nodes is the commonest presentation followed by laryngeal TB. 3) Fine needle aspiration cytology (FNAC) is a reliable and easy way to diagnose TB. However, newer diagnostic tests will increase the yield of positive cases and should be used whenever required. 4) In the larynx, the vocal cords were the commonest site affected and laryngeal TB need not be associated with lung TB or positive sputum always. 5) Patients who have TB of head and neck must be investigated to exclude pulmonary or systemic TB. 6) In cases of previous or subsequent TB infection, culture and drug sensitivity is indicated to reduce the problem of multiple drug resistance. PMID- 16360823 TI - Endoscopic sinus surgery for the treatment of chronic rhinosinusitis in patients with postirradiated nasopharyngeal carcinoma. AB - PURPOSE: Our purpose was to evaluate the efficacy and safety of endoscopic sinus surgery (ESS) in patients with nasopharyngeal carcinoma (NPC) after irradiation. MATERIALS AND METHODS: A retrospective study of postirradiated NPC patients who received ESS for the treatment of chronic rhinosinusitis in Taichung Veterans General Hospital. Symptoms, endoscopic findings, and computed tomography were used to evaluate the efficacy of ESS in these patients, and the safety was evaluated by intraoperative findings and complications. RESULTS: Since 1996, 10 postirradiated NPC patients with chronic rhinosinusitis have been treated by ESS and followed up for at least 2 years. The interval between radiotherapy and ESS was from 4 months to 16 years. After surgery, 7 patients felt improved except for one patient who died of tumor recurrence and computed tomography scores changed from 6.6 to 3.7, although prolonged nasal crusting was observed in most patients by endoscopy. One patient was complicated with cerebrospinal fluid leakage and the dura defect was immediately repaired without any sequelae. CONCLUSIONS: This study shows that ESS is effective for the treatment of chronic rhinosinusitis in postirradiated NPC patients if they failed medical treatment. PMID- 16360824 TI - Inverted papilloma-like sinonasal epithelial hyperplasia, overshadowing underlying sinonasal fibrosarcoma: a diagnostic pitfall. AB - Proliferating invaginations of the sinonasal epithelium, simulating inverted papillomas, have been mentioned once in the literature as a reactive phenomenon overlying sinonasal fibrosarcomas. These proliferations may be so marked as to make the distinction from inverted papilloma virtually impossible. The possible result is that the examining pathologist's attention might be drawn exclusively to the epithelial component, thus causing him to overlook, especially in a small biopsy, a rather bland but distinctive sarcomatoid component which is located underneath. We report such a case of marked endophytic proliferation of sinonasal epithelium which was interpreted, in the preoperative biopsy, as an inverted papilloma with dense fibrous stroma which later, in the surgical specimen, was found to be a part of a submucosal fibrosarcoma. The diagnostic pitfall is discussed. PMID- 16360825 TI - Malignant hypercalcemia associated with a parathyroid macrocyst and the early genesis of a giant cell tumor. AB - Parathyroid cysts rarely cause primary hyperparathyroidism. In most cases, the resultant hypercalcemia is mild and detected before any significant skeletal disease develops. We report a patient with severe hypercalcemia, a synchronous brown tumor (osteitis fibrosa cystica) of the maxilla, and a large benign functional parathyroid cyst. The unusual patient presentation and management are described and illustrated. The pertinent literature is reviewed. PMID- 16360826 TI - Epithelial-myoepithelial carcinoma in the base of the tongue: a case report. AB - OBJECTIVES: To report a rare case of epithelial-myoepithelial carcinoma (EMC) of a minor salivary gland originating in the base of the tongue, which is a highly unusual location. STUDY DESIGN: This is a case report and a review of literature. METHODS: A 76-year-old man was referred to our outpatient clinic with a 6-month history of a sensation of a foreign body in the pharynx. Endoscopic examination revealed a tumorous lesion 40 x 20 mm in the base of the tongue on the left side. A biopsy was performed, and the lesion was histologically identified as squamous cell carcinoma. RESULTS: The patient underwent subtotal glossectomy, bilateral neck dissection, and reconstruction with a microvascularized rectus abdominis myocutaneous flap. The final diagnosis based on the histological and immunohistochemical findings was EMC. Because neither histological evidence of residual tumor in the surgical margin nor neck metastases were found, radiotherapy was not performed. No local recurrence or distant metastases have been found 19 months post-surgery. CONCLUSION: We believe this report is the first description of EMC originating in the base of the tongue. We conclude that a wide surgical excision with a clear margin and close, prolonged follow-up are recommended for the treatment of EMC. PMID- 16360827 TI - Radioguided neck dissection in recurrent metastatic papillary thyroid carcinoma. AB - Although radioguided surgery has been used for the excision of sentinel nodes in breast cancer and melanoma, sparse literature exists describing its use in thyroid cancer. We report a 69-year-old patient with a previous total thyroidectomy and lymph node dissection for papillary carcinoma who was subsequently found to have recurrent metastatic disease. After a therapeutic dose of radioactive iodine, a hand-held gamma-probe was used to selectively dissect the neck. The patient was offered radioguided revision neck dissection to remove the disease using residual radioactivity of the original therapeutic iodine 131 dose. Our case report seeks to demonstrate a recent example of our use of the gamma-probe in radioguided surgical excision of recurrent metastatic papillary thyroid carcinoma. PMID- 16360828 TI - Giant cell tumor at the lateral skull base. AB - Giant cell tumor is a bony tumor which primarily occurs in third to fourth decade after skeletal maturation. Usually, this tumor arises in the distal end of long bone and is quite rare in the head and neck region. We had a elderly female patient with giant cell tumor at lateral skull base of the left temporal bone without classic symptoms. Team approach with neurosurgeon was used to eradicate the tumor. The detail of this case in conjunction with treatment of giant cell tumor was reported while reviewing pertinent papers. PMID- 16360829 TI - Cogan's syndrome: a cause of progressive hearing deafness. AB - In 1934 Morgan and Baumgartner first described a nonsyphilitic interstitial keratitis (IK) associated with vestibuloauditory dysfunction (Morgan RF, Baumgartner CF, Menier's disease complicated by recurrent interstitial keratitis. Excellent result following cervial ganglionectomy. West J Surg 1934;42:628). Cogan was the first to describe this syndrome as a clinical entity with the report of 5 additional cases in 1945 (Cogan DA: Syndrome of nonsyphilitic interstitial keratitis and vestibuloauditory symptoms. Arch Ophthalmol 1945;33:144-9). Since that time, more than 100 cases of Cogan's syndrome (CS) have been in the literature. Cogan's syndrome is a rare clinical disease, which primarily affects young adults; however, published reports range from 2.5 to 60 years for age of onset (Kundell Sp, HD Ochs: Cogan's syndrome in childhood. J Pediatr 1980;97:96-8). This disease primarily affects whites and is believed not to be hereditary. Typical CS is characterized by IK and vestibuloauditory dysfunction. The IK usually occurs with sudden onset and is characterized by photophobia, lacrimation, and eye pain. The vestibuloauditory dysfunction is usually bilateral, presenting with tinnitus, sensorineural hearing loss, and acute episodes of vertigo. Atypical CS presents with significant inflammatory eye disease (ie, scleritis, episcleritis, retinal artery occlusion, choroiditis, retinal hemorrhages, papilloedema, exophthalmos, or tenonitis) with or without IK (Laryngoscope 1960;70:447-9). In this report, we describe a typical case of CS, in which the hearing loss was unresponsive to corticosteroid therapy. PMID- 16360830 TI - Epidermoid formation: the potential precursor of congenital cholesteatomas. PMID- 16360832 TI - Characteristics and predictors for non-vaccination with PPV in the elderly population. PMID- 16360833 TI - Dendritic cells expand Epstein Barr virus specific CD8+ T cell responses more efficiently than EBV transformed B cells. AB - Adoptive transfer of Epstein Barr virus (EBV) specific cytotoxic T lymphocytes (CTLs) has been successfully applied in the treatment of EBV associated post transplant lymphoproliferative disease (PTLD). In most studies EBV transformed B cells (LCLs) have been used for the induction of EBV specific T cell lines. Application of this approach to other EBV associated tumors is difficult, because LCLs focus T cell expansion toward immunodominant EBV antigens that are not expressed in EBV associated Hodgkin's lymphoma and nasopharyngeal carcinoma. Therefore, we compared dendritic cells (DCs) with LCLs for CD8+ T cell stimulation against dominant and subdominant EBV antigens. DCs expanded tenfold more EBNA3A and LMP2 specific CD8+ T cells than LCL and also stimulated EBV specific CTL from PTLD patients. Both, DCs and LCLs stimulations led to the expansion of high affinity T cells, capable to target EBV transformed B cells. While LCLs and DCs expressed MHC class I and II products at similar levels, DCs showed a higher expression of costimulatory and adhesion molecules. This resulted in more efficient T cell conjugate formation with DCs than with LCLs. We propose the use of DCs for stimulation of EBV specific T cells in active or passive immunotherapy of EBV associated malignancies. PMID- 16360834 TI - Direct ex vivo detection of HLA-DR3-restricted cytomegalovirus- and Mycobacterium tuberculosis-specific CD4+ T cells. AB - In order to detect epitope-specific CD4+ T cells in mycobacterial or viral infections in the context of human class II major histocompatibility complex protein human leukocyte antigen (HLA)-DR3, two HLA-DR3 tetrameric molecules were successfully produced. One contained an immunodominant HLA-DR3-restricted T-cell epitope derived from the 65-kDa heat-shock protein of Mycobacterium tuberculosis, peptide 1-13. For the other tetramer, we used an HLA-DR3-restricted T-cell epitope derived from cytomegalovirus (CMV) pp65 lower matrix protein, peptide 510 522, which induced high levels of interferon (IFN)-gamma-producing CD4+ T cells in three of four HLA-DR3-positive CMV-seropositive individuals up to 0.84% of CD4+ T cells by intracellular cytokine staining. In peripheral blood mononuclear cells from M. tuberculosis-exposed, Mycobacterium bovis bacille Calmette-Guerin (BCG)-vaccinated, or CMV-seropositive individuals, we were able to directly detect with both tetramers epitope-specific T cells up to 0.62% and 0.45% of the CD4+ T-cell population reactive to M. tuberculosis and CMV, respectively. After a 6-day culture with peptide p510-522, the frequency of CMV-specific tetramer binding T cells was expanded up to 9.90% tetramer+ CFSElow (5,6 carboxyfluorescein diacetate succinimidyl ester) cells within the CD4+ T-cell population, further confirming the specificity of the tetrameric molecules. Thus, HLA-DR3/peptide tetrameric molecules can be used to investigate HLA-DR3 restricted antigen-specific CD4+ T cells in clinical disease or after vaccination. PMID- 16360835 TI - A subpopulation of human basal keratinocytes has a low/negative MHC class I expression. AB - By means of flow cytometry, we demonstrate that a subpopulation of cells in the basal layer of the human epidermis has undetectable levels of major histocompatibility complex (MHC) class I molecule. The percentage of MHC class I negative cells in the basal layer ranged 0.5%-2%. MHC class I-negative cells were characterized by small size (low forward scatter) and low granularity (low side scatter). Upon culturing of MHC class I-negative cells, increase of MHC class I expression was observed. This expression was lower than the expression of MHC class I molecule both in cultured MHC class I-positive cells and in ex vivo keratinocytes. Furthermore, stimulation of MHC class I-negative keratinocytes with interferon gamma (IFN-gamma) did not bring about further increase in MHC class I expression. MHC class I-negative cells were identified as keratinocytes as they expressed keratin 14 and formed keratinocyte colonies in vitro. PMID- 16360836 TI - Allogeneic MHC class I molecules with numerous sequence differences do not elicit a CTL response. AB - CD8+ T cell-mediated alloreactivity is generally believed to involve recognition of the alpha1/alpha2 domains of donor-type class I MHC molecules as well as the peptides they bind. Using the CTLp assay outcome as a parameter for the induction of alloreactivity, we have retrospectively surveyed 80 haematopoietic stem cell donor/patient pairs that feature a range of allelic differences at single HLA-A, B, and -C loci in an attempt to probe the predictive value of such mismatches. In contrast to the expectation that greater degree of allelic disparity would lead to more alloreactivity, we found that in a substantial number of cases, class I MHC molecules with numerous sequence differences did not elicit an allogeneic CTL response. We propose that in generating a T cell repertoire with a sufficiently narrow responsive for self-MHC, positive thymic selection limits the capacity to recognize allogeneic MHC molecules whose structure and sequence have diverged extensively. These findings are important for donor and patient MHC matching strategies and our understanding of T cell-MHC interaction after haematopoietic stem cell transplantation. PMID- 16360837 TI - Microparticle-free placentally derived soluble factors downmodulate the response of activated T cells. AB - Soluble placental factors may have immunoregulatory properties and have been demonstrated to inhibit T-lymphocyte proliferation in vitro. On the other hand, placentally derived syncytiotrophoblast microparticles and crude placental homogenates have been demonstrated to inhibit proliferation of mixed lymphocytes in vitro. Because previous studies on placentally derived soluble factors may have been contaminated by the presence of trophoblast-derived microparticles, we prepared microparticle-free placental supernatants. Such supernatants reduced the activation response of T cells, as well as their proliferation and the production of cytokines such as interleukin-2 and interferon gamma, in a dose-dependent manner. This reduction in T-cell proliferation does not appear to be caused by indoleamine 2,3-dioxygenase (IDO) because it was not reversed by the addition of L-tryptophan or an inhibitor of IDO (1-methyl-DL-tryptophan). No evidence was found for the presence of IDO in these supernatants when we used a biochemical assay measuring tryptophan catabolism. We conclude that the placenta produces currently unknown soluble factors that reduce T-cell activation, proliferation, and cytokine production. PMID- 16360838 TI - Expansion of humoral donor-specific alloreactivity after renal transplantation correlates with impaired graft outcome. AB - IgG human leukocyte antigen (HLA) antibodies were investigated retrospectively after transplantation in 264 primary renal graft recipients. All patients who developed de novo donor-specific antibodies (DSA) and non-DSA (NDSA) (n = 40, 15.1%) were divided into two groups. Group A consisted of patients (n = 20) with stable good graft function (GGF), and group B consisted of patients (n = 20) who developed rejection and/or graft failure (R/GF). DSA were detected in 23 patients (57.5%). Expansion of humoral alloreactivity with the presence of DSA to more than one graft mismatched antigens and coexistence of HLA class I and II DSA were significantly correlated only with R/GF (p = 0.01). Limitation of alloreactivity to one graft-mismatched antigen was detected mainly in patients with GGF. HLA-DQ DSA alone was found only in patients with GGF (p = 0.1). No significant differences were found between the two patient groups with NDSA. Expansion of the humoral alloreactivity to more than one graft molecule in renal transplant recipients identifies patients at high risk of rejection or graft failure. Limitation of humoral alloreactivity to one graft antigen perhaps associates the presence of regulatory mechanisms with GGF only in specific cases. PMID- 16360839 TI - HLA antibody identification with single antigen beads compared to conventional methods. AB - Single antigen (SA) beads coated with Class I HLA antigens from recombinant cells lines were tested with 170 mouse monoclonal antibodies (mAbs). The HLA specificities of all mAbs were previously determined by the cytotoxicity assay (CDC). There were 100 mAbs which produced the expected reactions with the SA beads, indicating that the SA beads coated with the antigens had reacted properly. Sixty one mAbs were positive with one or more antigen(s) that shared unique amino acids (aa) possibly constituting a common epitope. Single antigen beads were then tested on 58 alloantisera analyzed by 63 laboratories of the UCLA serum exchange (UCLA-SE). Many specificities detected by the single antigen beads were missed by the laboratories employing conventional methods. Most of the missed specificities were of lower frequency, although in some instances, even common specificities were missed. These findings have important implications regarding the use of specificities to predict positive crossmatch, to selecting platelet donors for highly sensitized recipients, and analysis of sera for donor specific antibodies. PMID- 16360840 TI - TAP1 and TAP2 gene polymorphisms and HLA-TAP haplotypes in Koreans based on 90 families. AB - We have investigated the polymorphism of TAP genes and the distribution of human leukocyte antigen (HLA)-TAP haplotypes in 90 Korean families (180 parents and 233 children), previously typed for HLA class II alleles. TAP1 (codons 333 and 637) and TAP2 (codons 379, 565, 577, 651, 665, and 687) typings were carried out by use of polymerase chain reaction-restriction fragment length polymorphism method. For TAP1, four alleles (gene frequency: A 81.9%, B 15.0%, C 2.5%, D 0.5%) and for TAP2, nine alleles (A1 31.7%, A2 14.2%, B 32.5%, Bky2 6.1%, C 6.9%, D 2.5%, E 3.9%, G 0.8%, and H 1.4%) were detected. Sixteen different TAP1-TAP2 haplotypes were observed with frequencies > 0.5%, and we found that significant linkage disequilibrium was present between TAP1 and TAP2 loci (p < 0.001). When HLA-DRB1 DQB1 haplotypes were extended to TAP1 and TAP2 loci, much diversification of haplotypes was observed: 26 different DRB1-DQB1 haplotypes (frequencies > 0.8%) formed 90 different extended haplotypes. Eight recombinant haplotypes between DQB1 and DPB1 loci were observed, and most (seven of eight) of the recombinations occurred between TAP2 and DPB1 loci. These results add more evidence to the view that recombination hot spot is present within and around TAP gene region. PMID- 16360841 TI - No association between transmembrane protein-tyrosine phosphatase receptor type C (CD45) exon A point mutation (77C>G) and idiopathic dilated cardiomyopathy. AB - To investigate whether a 77C>G polymorphism in exon A of the CD45 gene causing a variant CD45RA expression pattern is associated with the development of idiopathic dilated cardiomyopathy (DCM), we studied a total of 414 individuals (104 patients and 310 controls). CD45RA expression pattern on lymphocytes was examined by flow cytometric analysis and subsequently the CD45 77C>G polymorphism was genotyped by polymerase chain reaction-allele specific restriction enzyme analysis (PCR-ASRA). We found 5 patients and 8 control individuals displaying the variant CD45RA expression pattern. All identified individuals carried the heterozygous CD45 77C>G polymorphism. The frequency of the 77G allele in the patient group was 2.4%, which was not significantly different from 1.3% found in the control group (p=0.327). In conclusion, the data of this preliminary study could not reveal any association between the CD45 77C>G polymorphism and susceptibility to idiopathic DCM in a German population. PMID- 16360842 TI - Nomenclature for factors of the HLA system, update January 2005. PMID- 16360843 TI - Nomenclature for factors of the HLA system, update February 2005. PMID- 16360844 TI - Nomenclature for factors of the HLA system, update March 2005. PMID- 16360845 TI - Nomenclature for factors of the HLA system, update April 2005. PMID- 16360846 TI - Nomenclature for factors of the HLA system, update May 2005. PMID- 16360847 TI - Nomenclature for factors of the HLA system, update June 2005. PMID- 16360848 TI - Radiopacity of dental materials using a digital X-ray system. AB - OBJECTIVES: Radiopacity is a desirable property for most intra-oral materials. There are established ISO and ANSI/ADA protocols for determining radiopacity using film-based radiography. However, these methods are not always followed by researchers. This study aims to adapt those procedures by using digital radiography, a simplified stepwedge, and examine the effects of target distance and exposure time choice. METHODS: One millimetre thick samples of three dental materials were prepared by placing the materials into a 1.00 mm thick washer sandwiched between two glass slides. The samples were digitally radiographed alongside a stepwedge of aluminum alloy 1100 with an X-ray unit at 70 kVp using five different target distance/exposure time combinations. For each combination, the grey scale values of various thicknesses of the stepwedge were converted into absorbencies and plotted against their thickness. These plots were then linearly regressed in order to correlate absorbance with a thickness of aluminum for each target distance/exposure time combination. The absorbencies of each sample were then converted into radiopacities using these correlations. RESULTS: The correlations between the absorbance of the stepwedge and its thickness were highly linear. This linearity allows the correlation to be accurately deduced from fewer data points than required by the ISO and ANSI/ADA protocols. Varying exposure time did not significantly affect the mean radiopacity measured at a target distance of 30 cm. Varying the target distance did not significantly affect the measured radiopacity as long as the samples were properly exposed. SIGNIFICANCE: A simplified, consistent digital method for determining radiopacity is presented. PMID- 16360849 TI - Effect of material properties on stresses at the restoration-dentin interface of composite restorations during polymerization. AB - BACKGROUND: Numerous analyses for the shrinkage stress in the adhesive resin based composite restorations mostly rely on numerical models. However, various finite element studies have inherent difficulties and inconsistencies associated with the use of different anatomy (tooth and restoration), boundary conditions (root and interfaces) and shrinkage models. As a consequence many numerical results remain inconclusive. OBJECTIVE: The objective of this paper is to develop a simplified analytical model of shrinkage stress and investigate effects of material properties of the restorative material, size of the restoration and volumetric shrinkage on the magnitude of the shrinkage stress in the vicinity of the dental-restoration interface. METHODS: The model is based on the following assumptions. The geometry is axisymmetric; all materials are linear-elastic; and the polymerization of the restoration material results in uniform volume shrinkage. An application of compatibility conditions leads to the system of five linear algebraic equations to five unknown variables, which can be easily resolved using standard techniques. RESULTS: An explicit equation for the tensile stress at the interface was obtained. It was shown that higher Young's modulus, Poisson's ratio and volume shrinkage of the restorative material normally lead to larger tensile stress at the interface, which increases the risk of debonding. The results obtained based in this work, in general, are in a good agreement with published results of finite element studies. SIGNIFICANCE: The model allows comparison of different adhesive restorative materials with respect to the fracture risk of the interface induced by the development of the shrinkage stress at the restoration-dentine interface during polymerization. The model can be used to validate more sophisticated computational models as well as to conduct various optimization studies and preliminary assessments of fracture risk. PMID- 16360851 TI - Neck dissections for oral/head and neck cancer: 1906-2006. PMID- 16360852 TI - The use of temporal polyethylene implant after temporalis myofascial flap transposition: clinical and radiographic results from its use in 21 patients. AB - PURPOSE: The use of temporalis myofascial flap (TMF) as a pedicled flap in craniofacial reconstructive surgery is well established. The transposition of temporalis muscle results in a large hollowing of the temporal fossa that leaves the patient with a cosmetic impairment. Reconstruction of this donor site deformity is desirable. One of the established reconstructive techniques is the use of a prefabricated porous high-density polyethylene (HDPE) temporal implant. In order to evaluate results from its use, we retrospectively reviewed a series of 21 consecutive patients. MATERIALS AND METHODS: From October 1999 to October 2004, 21 patients (7 men and 14 women) aged 32 to 85 years (mean, 65) had their surgical defects reconstructed with the use of a TMF. The majority of patients (15 of 21) had squamous cell carcinoma of the maxilla or the maxillary sinus. In 17 patients, the reconstructive procedure was performed simultaneously with the oncological resection, whereas in 4, a secondary reconstruction was performed. In 1 patient, bilateral TMFs were used to cover a total maxillectomy defect. Standard surgical approach was used in all patients during TMF elevation. The temporal defect was reconstructed with the use of a prefabricated sterile HDPE implant (Medpor; Porex Surgical Inc, College Park, GA). Fixation of the implant to the recipient infratemporal fossa was performed with black silk sutures (in 2 patients) or titanium miniscrews (in 19 patients). The manufacturer's instructions for the placement of the implant were followed in all cases. One of the 21 operated patients preoperatively received radiotherapy (RT). Of the remaining 20 patients, 5 underwent postoperative RT. RESULTS: Eighteen patients are alive and free from disease. One died during the perioperative period from myocardial infarction and 2 more from locoregional recurrence of their disease, 18 and 27 months postoperatively. In all 21 patients, the placement of the Medpor temporal implant was successful and no immediate or perioperative complications resulting from its use were encountered, giving an overall success implantation rate of 100%. Follow-up ranged from 9 to 70 months (mean, 39). The condition of the implant was evaluated with computed tomography in 18 of the 21 patients as part of the standard postoperative assessment. Radiographic results of the recipient site did not reveal any abnormalities. In 7 patients, the contour of the HDPE implant could be manually palpated, and in 3, it could be seen to protrude subcutaneously. Esthetic results were judged satisfactory from all patients. The hemicoronal skin flap healed uneventfully in all patients and did not cause a visible scar even to bald male patients. CONCLUSIONS: The reconstruction of the temporal defect after TMF transposition with the use of a Medpor temporal implant is an easy and safe method. The implant does not seem to cause any tissue reaction, and long-term functional and esthetic results are excellent. When properly used and the relevant manufacturers' instructions are carefully followed, the success rate of the method is extremely high. PMID- 16360853 TI - Combined push-pull distraction for correction of syndromic midfacial hypoplasia. AB - PURPOSE: Gradual midfacial advancement, applying the principle of distraction osteogenesis, reduces the restriction of the soft tissues and results in bony consolidation without need for grafting. The midface can be distracted by either pushing it forward, using semiburied devices, or pulling it forward by a rigid external device. For each method there are inherent technical problems, such as controlling the vector of movement, symmetry of advancement, and differential movement of the upper/lower face. We have used a combination of the 2 methods, called "push-pull," in an effort to control the distraction process. The purpose of this paper is to describe our push-pull distraction technique and summarize our early experience. MATERIALS AND METHODS: Ten patients (5 males and 5 females) with a mean age of 11 years 2 months underwent midfacial advancement using push pull distraction. Two orthodontists, blinded for landmark identification, traced preoperative and postoperative cephalograms and determined linear and angular measurements of midfacial position. A Student t test was used to assess differences between the cephalometric measures on the 2 radiographs. Interexaminer reliability was calculated by an intraclass correlation coefficient. RESULTS: Postdistraction cephalograms were taken a mean of 10 months (range, 3 to 20 months) after removal of the devices. Patients exhibited improvement at all levels of the midface after distraction. There was a statistically significant sagittal advancement from the infraorbital rim to dentoalveolus. The central midface was sufficiently advanced as shown by an improved convexity, nasolabial angle, and upper labial protrusion. There were no significant differences between examiners for any of the measurements in this study. CONCLUSIONS: Push-pull distraction permits 1) equal movement at both the upper and lower facial levels, 2) advancement of the central midface, and 3) symmetric movement of the zygomaticomaxillary complexes. This method also provides a backup, in case one device malfunctions. In combination, the advantages of each device are additive; whereas the weaknesses are not. The push pull technique is a practical method for midfacial distraction until a better single device is fabricated. PMID- 16360854 TI - The use of biodegradable plates and screws to stabilize facial fractures. AB - PURPOSE: The purpose of this preliminary retrospective study was to review the demographics and outcome of patients with a variety of facial fractures that were stabilized with PL bone plates and screws. MATERIALS AND METHODS: The records of 295 consecutive patients with facial fractures treated by open reduction and internal fixation, performed by the author from 2001 through 2004, were retrospectively reviewed. Patients were selected to receive biodegradable fixation on the basis of mechanism of injury, the degree of bony displacement demonstrated on clinical and radiographic examination, patient age, and fracture pattern or location. Outcome measures such as infection, non-union, and mal-union were identified and subjectively assessed. Descriptive statistics were recorded and analyzed. RESULTS: Two hundred eighty-one patients met the criteria for inclusion in the study with follow-up of 3 weeks to 3 years. Fifty-nine (21%) patients were identified as having received biodegradable plates and screws. All patients eventually went on to satisfactory healing with favorable restoration of form and function. Complications occurred in 16 patients overall (6%). Of these, 2 patients were treated with resorbable plates and screws; 1 patient with a zygomatico-maxillary complex fracture developed a sterile abscess that presented 1 year postoperatively and responded to local measures. Another patient with a Le Fort I fracture developed an anterior open bite necessitating Le Fort I osteotomy for correction. CONCLUSIONS: Favorable healing can be observed through the use of biodegradable PL plates and screws to stabilize selected midface fractures in patients of all ages, as well as mandible fractures in early childhood. PMID- 16360855 TI - Self-reinforced biodegradable screw fixation compared with titanium screw fixation in mandibular advancement. AB - PURPOSE: This report compares the skeletal stability and treatment outcomes of 2 similar cohorts undergoing bilateral sagittal osteotomies of the mandible for advancement. The study groups included patients stabilized with 2-mm self reinforced polylactate (PLLDL 70/30), biodegradable screws (group B), and 2-mm titanium screws placed in a positional fashion (group T). MATERIALS AND METHODS: Sixty-nine patients underwent bilateral sagittal osteotomies of the mandibular ramus for advancement utilizing an identical technique. There were 34 patients in group B and 35 patients in group T. Each patient had preoperative, immediate postoperative, splint out, and 1-year postoperative cephalometric radiographs available for analysis. The method of analysis and treatment outcomes parameters are identical to those previously used. Repeated measures analysis of variance was performed with means of fixation as the between-subject factor and time as the within subject factor. The level of significance was set at .01. RESULTS: There were no clinical failures in group T and a single failure in group B. The average difference in stability between the groups is small and subtly different at the mandibular angle. The data documented similarity of the postsurgical changes in the 2 groups with the only statistically significant difference being the vertical position of the gonion (P < .001) and the mandibular plane angle (P < .01) with greater upward remodeling at gonion in group T. CONCLUSIONS: Two-mm self-reinforced PLLDL (70/30) screws can be used as effectively as 2-mm titanium screws to stabilize the mandible after bilateral sagittal osteotomies for mandibular advancement. The difference in 1-year stability and outcome is minimal. PMID- 16360856 TI - Bioresorbable poly-L/DL-lactide (P[L/DL]LA 70/30) plates are reliable for repairing large inferior orbital wall bony defects: a pilot study. AB - PURPOSE: The purpose of this study was to share our clinical experience on the use of bioresorbable poly-L/DL-lactide implants (P[L/DL]LA) 70/30 (PolyMax; Synthes, Oberdorf, Switzerland) to repair, large (> or =2 cm2), inferior orbital wall defects and to evaluate whether P(L/DL)LA 70/30 implants adequately support the orbital soft tissue contents. PATIENTS AND METHODS: Thirteen patients who suffered orbital blowout fractures, with > or =2 cm2 bony defects in the inferior orbital wall, took part in the study. The inferior orbital wall was explored via subconjunctival approach. After repositioning of orbital content, each inferior orbital wall was reconstructed using a round plate of P(L/DL)LA 70/30. Computed tomography and magnetic resonance imaging coronal sections were undertaken before the operation and 2 and 36 weeks postoperatively. RESULTS: The magnetic resonance imaging studies showed no abnormal tissue foreign body reactions in the orbital region. The material showed adequate strength to stabilize bone segments during the critical period of bone healing. The bone healing seems to take place along the bone fragments. The clinical outcome was excellent in 11 of the 13 cases (85%). At the end of the study, only one patient had mild enophthalmos. CONCLUSIONS: Bioresorbable P(L/DL)LA 70/30 implants are safe and reliable for the repair of large defects (> or =2 cm2) in the inferior orbital wall. It seems that this is the first reported biodegradable material, in the literature, to promote bone healing along the bone fragments of the inferior orbital wall. PMID- 16360857 TI - Motorcycle-related maxillofacial injuries among Nigerian intracity road users. AB - PURPOSE: We sought to determine the etiology and pattern of motorcycle-related maxillofacial injuries among intracity road users in Nigeria. PATIENTS AND METHODS: We conducted a prospective review of all patients presenting at the maxillofacial surgery units of the Obafemi Awolowo University Teaching Hospital Ile-Ife and Usmanu Dan Fodiyo University Teaching Hospital Sokoto with motorcycle related injuries. RESULTS: One hundred seven patients were included in the study. A significant male preponderance was observed (P = .002). Overall, the peak age incidence was 20 to 29 years, whereas female patients had a peak age incidence of 10 to 19 years. Victims were mostly riders (50.5%) but none of them was a female. While 8.4% of accidents occurred on motorcycles with more than one pillion passenger, none of the victims reported using a crash helmet. Mechanism of accident was mostly head-on collision (39.2%). The injuries sustained were predominantly soft tissue injuries or in combination with bone injury. Bone and dental injuries were attributed mostly to falls. Facial bone injuries occurred mostly in the mandible (64 of 104). A symmetric distribution of injuries was observed in the upper, middle, and lower thirds of the face, but the middle third had the highest injury sites while the upper third had the least. CONCLUSIONS: Motorcycle-related maxillofacial injuries among Nigerians affect males predominantly. The injuries sustained have a symmetric distribution and were mostly soft tissue injuries in isolation or in combination with bony injuries. The middle third of the face is most vulnerable. Mandatory use of safety helmets and education of cyclists on the appropriate number of pillion passengers are imperative to minimize the morbidity and mortality associated with motorcycle accidents. PMID- 16360858 TI - A 5-year retrospective study of zygomatico-orbital complex and zygomatic arch fractures in Sao Paulo State, Brazil. AB - PURPOSE: This descriptive analytical study evaluated the cause, type, incidence, demographic, and treatment data of zygomatico-orbital complex and zygomatic arch fractures managed at our program during a 5-year period. PATIENTS AND METHODS: A total of 1,857 charts of patients presenting maxillofacial trauma and treated by the Division of Oral and Maxillofacial Surgery, from the Piracicaba Dental School (UNICAMP), were reviewed. A total of 371 patients presented with zygomatico orbital complex and/or zygomatic arch fractures. Patients with isolated orbital fractures were excluded. The patients ranged in age from 2 to 88 years. Epidemiological data including age, gender, etiology, site of injury, and details of treatment were collected and tabulated in Excel software (Microsoft). All maxillofacial injuries were assessed and treated by residents and staff surgeons. Other concomitant bodily injuries were treated by pertinent consulting specialists. RESULTS: The majority of fractures were sustained by males between 21 and 40 years old, usually resulting from falls, altercations, and bicycle accidents. Unilateral fractures of zygomatico-orbital complex were present in 266 patients (71.69%). Bilateral fractures were found in 22 cases (5.93%). Unilateral zygomatic arch fractures occurred in 39 cases (10.51%). A total of 210 patients (56.60%) did not receive surgical treatment. Surgical intervention was done in 161 patients (43.40%). The surgical treatment was conducted using the approach as reported by Ellis and Kittidumkerng in 1996. A complication incidence of 6.47% was observed, which is compatible with related articles in the literature. CONCLUSIONS: The study revealed interesting features about etiology, demonstrated by the high occurrence of bicycle accidents. The treatment protocol proposed to the patients showed adequate results as evidenced by low complication frequency. The findings of this study bring information about an area in Sao Paulo State, Brazil, that is valuable in reaffirming previously established trends and identifying new patterns of disease frequency. PMID- 16360859 TI - Intrarater reliability in the ultrasound diagnosis of medial and lateral orbital wall fractures with a curved array transducer. AB - PURPOSE: The aims of the study were to document the effectiveness of ultrasound (US) in diagnosing orbital wall fractures when compared with computed tomography (CT) and to measure the intraobserver reliability of US using a curved array transducer. MATERIALS AND METHODS: From December 2003 to March 2004, 13 patients with the clinical diagnosis of an orbital trauma were investigated prospectively by CT (reference) and 2 US investigators. Both orbits were investigated. Sensitivity, specificity, accuracy, and positive and negative predictive value were calculated. The statistical difference between the 2 US investigators was calculated by a chi-square test. The interrater reliability was calculated using the lambda coefficient. Values below 0.4 represent poor reliability, between 0.4 and 0.75 represent fair to good reliability, and a score > 0.75 is graded as excellent reliability. RESULTS: The comparison of the results of the 2 US investigators by the chi-square test showed P values of .385 for the medial orbital wall and .638 for the lateral orbital wall, which shows no significant difference. The lambda-value for the investigation of the medial orbital wall reached 0.429, 0.714, and 0.750. The lambda-value for the investigation of the lateral orbital wall yielded 0.647, 0.750, and 0.882. These values show a good and excellent inter-rater reliability. CONCLUSION: The US investigation does not yet reach the diagnostic quality of CT. US could be a helpful diagnostic imaging tool in cases with clear clinical symptoms. The results of the current study and the previously published results imply that US has the potential to reach the same diagnostic quality as CT in the future, but further studies must be performed to improve the diagnostic quality of the method. PMID- 16360860 TI - Maxillary stability following Le Fort I osteotomy in combination with sagittal split ramus osteotomy and intraoral vertical ramus osteotomy: a comparative study between titanium miniplate and poly-L-lactic acid plate. AB - PURPOSE: The purpose of this study was to compare changes in maxillary stability after Le Fort I osteotomy with titanium miniplate and poly-L-lactic acid (PLLA) plate (Fixsorb-MX; Takiron Co, Osaka, Japan). PATIENTS AND METHODS: The subjects were composed of 47 Japanese patients with diagnosed jaw deformity: 24 underwent Le Fort I osteotomy and sagittal split ramus osteotomy (SSRO); and 23 underwent Le Fort I osteotomy intraoral vertical ramus osteotomy without internal fixation. Each group was divided into titanium plate and PLLA plate groups. Time course changes between plate groups were compared using lateral and posteroanterior cephalography. RESULTS: Significant differences were identified between titanium plate and PLLA plate groups in A point after Le Fort I osteotomy and SSRO (P < .05). Significant differences existed between titanium plate and PLLA plate groups in vertical component of posterior nasal spine after Le Fort I osteotomy in both combinations with SSRO and intraoral vertical ramus osteotomy (P < .05). However, no significant differences were identified in measurements on posteroanterior cephalography. CONCLUSION: These results suggest a slight tendency for vertical impaction after Le Fort I osteotomy both in combination with SSRO and intraoral vertical ramus osteotomy with PLLA plates, although differences in time course changes were not clinically apparent, and normal occlusion was established in all patients. PMID- 16360861 TI - Predictive factors for the histologic response to preoperative radiotherapy in advanced oral cancer. AB - PURPOSE: Preoperative radiotherapy and chemotherapy are important in treating advanced oral cancer. We attempted to elucidate predictive factors for the histologic response to preoperative radiotherapy or chemoradiotherapy. PATIENTS AND METHODS: Forty-three patients with locally advanced cancer of the oral cavity and oropharynx were treated preoperatively with radiotherapy (50 Gy); surgery included modification of the resected area to preserve organ function. RESULTS: Hemoglobin level and histologic differentiation were independent factors significantly associated with the histologic response by multivariate analysis. Locoregional failure occurred only in patients with poor histologic response, and there was a significant difference in the 5-year survival rate between patients with good and poor responses (84% vs 32%). Multivariate analysis indicated that the histologic response at the primary site and the number of metastatic lymph nodes were prognostic factors. CONCLUSION: We identified predictive factors for a good response to preoperative therapy and found a high survival rate in patients with a good response. PMID- 16360862 TI - The acute orbit: etiology, diagnosis, and therapy. AB - PURPOSE: Extension of dental abscesses to distant areas of the head and neck has been repeatedly reported in the medical literature. Subsequent involvement of the orbit still remains a rarity, resulting in protracted onset of therapy because of inaccurate diagnosis. Considering the possible lasting damage that can result from insufficient therapy, like blindness or even death, the need to extensively educate practicing physicians becomes evident. MATERIALS AND METHODS: With the help of a comprehensive review of the medical literature underlined with a clinical case, the etiology, diagnosis, and therapy of the acute orbit are highlighted with emphasis on the new imaging modalities as well as the broad spectrum antibiotics currently available on the market. RESULTS: Orbital infections of odontogenic origin are the rarest sequelae, with a prevalence of 1.3%. Correct diagnosis, adequate antibiotic therapy, and surgical drainage are the keys to success. CONCLUSION: The acute orbit continues to be a medical challenge. With the proposed diagnostic and therapy guidelines, this affliction can be identified and contained with a high degree of certainty. PMID- 16360863 TI - Radiographic evaluation of the fate of developing tooth buds on the fracture line of mandibular fractures. AB - PURPOSE: The goal was to suggest guidelines for the treatment of developing tooth buds located on the fracture line of mandibular fractures. PATIENTS AND METHODS: The long-term radiographic follow-up records of 28 patients with mandibular fractures involving 66 tooth buds were examined for the occurrence of abnormalities in development or eruption. The fates of the involved teeth were compared according to the fracture conditions and other factors, and the cause of the abnormalities was surveyed. RESULTS: Abnormal findings were observed in 30 of 66 developing teeth (45%); these included deficient root formation, abnormal bend of the root, nodule formation on the root, partial obliteration of the pulp cavity, impaction, growth arrest, and external resorption. No relationship was found between the presence of abnormalities and the condition of the fracture or the developmental stage of the tooth buds. However, infection, rotation of the tooth bud, and a surgical wire passing through the follicular space were associated with arrested growth and impaction. CONCLUSIONS: The tooth buds present on the fracture line should be preserved except in cases of infection, and careful attention should be paid to avoiding further injury to the tooth bud and the soft tissues of the follicle at the time of surgery. PMID- 16360864 TI - Intraocular pressure changes secondary to reduction of orbito-zygomatic complex fractures. AB - PURPOSE: To investigate intraocular pressure (IOP) changes in the involved eye during orbito-zygomatic complex fracture reduction, compared with the IOP of the eye on the uninjured side. PATIENTS AND METHODS: Twenty patients who presented with displaced, isolated unilateral orbito-zygomatic complex fractures were enrolled. IOPs were measured using a Tono-Pen XL (Medtronic Solan, Jacksonville, FL) in the involved eye and in the non-involved eye. Pressure was recorded at 5 time points (before general anesthetic induction, after induction, immediately after fracture reduction, post operation while still under general anesthetic, and 15 minutes post general anesthetic). RESULTS: There were no significant differences between the injured and control data at any time point. Average IOPs for the involved and control eye decreased after anesthetic induction; however, IOP increased in both eyes after fracture reduction. CONCLUSION: There is no statistical evidence of a difference between the IOP of the control and injured eyes. Routine IOP testing is not indicated during orbito-zygomatic fracture reduction. PMID- 16360865 TI - Surgical approaches to the infraorbital rim and orbital floor: the case for the subtarsal approach. PMID- 16360866 TI - Surgical approaches to the infraorbital rim and orbital floor: the case for the transconjunctival approach. PMID- 16360867 TI - Hypertension: classification, pathophysiology, and management during outpatient sedation and local anesthesia. PMID- 16360868 TI - The use of immediate bone grafting in reconstruction of clinically infected mandibular fractures: bone grafts in the presence of pus. AB - PURPOSE: Current approaches to the treatment of infected mandibular fractures include antibiotics, drainage, immobilization of the segments, and debridement followed by secondary bone grafting of residual defects once the infection is resolved and the wound healed. Over the past 30 years, the time from debridement to grafting has diminished from several months to a few weeks. We present our experience with a treatment model managing clinically infected fractures of the mandible with antibiotics, debridement, rigid internal fixation, and immediate autogenous bone grafting. MATERIALS AND METHODS: In this retrospective study, we present a series of 43 patients who demonstrated clinical/laboratory findings consistent with infection in one or more mandibular fractures (50 infected fractures). These patients underwent a combination of incision and drainage, fracture debridement, rigid internal fixation, and immediate bone grafting of the resulting defect in a single stage. Both transoral and transfacial approaches were used. RESULTS: Of the 50 fractures, 43 showed both resolution of infection and bony union of fractures with long-term follow-up of 2 months to 4 years. Four fractures developed recurrent infection but proved to have bony union and were successfully treated by hardware removal only. Three other patients were deemed failures with persistent infection, loss of graft, nonunion, and need for retreatment. Each of these patients was afflicted with underlying immunocompromise. CONCLUSIONS: Although careful patient selection is a must, immediate bone grafting of infected mandibular fractures, when used in conjunction with rigid internal fixation and appropriate intraoperative debridement, is an effective treatment modality which allows a single surgical procedure and dramatically shortens the course of treatment. PMID- 16360869 TI - Multiple teeth extractions in a patient with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), treated with ticlopidine. PMID- 16360870 TI - Intracranial migration of a halo pin during distraction osteogenesis for maxillary hypoplasia: case report and literature review. PMID- 16360871 TI - Loss of a sewing needle in the tongue during attempted tongue piercing: report of a case. PMID- 16360872 TI - An unusual bullet trajectory to the face. PMID- 16360873 TI - Dermatofibrosarcoma protuberans of the face: report of 2 cases and an overview of the recent literature. PMID- 16360874 TI - Giant mandibular bone defect: report of a case. PMID- 16360875 TI - OMFS practitioners who do not take emergency calls. PMID- 16360877 TI - Effects of specific and non-specific perceived control on blood pressure response in a stressful mental task. AB - Eighty students volunteered to participate in an experiment with the aim of evaluating the impact of the manipulation of self-efficacy and negative incentive values on systolic and diastolic blood pressure, and pulse pressure. The subjects were asked to solve a series of 15 mathematical problems after having been randomly assigned to one of the four experimental conditions that were generated by combining two levels of self-efficacy (high versus low), and two levels of negative incentive value (high versus low), contingent upon failing to properly perform the task. The subjects' perceived competence was also evaluated. The results are consistent with those obtained in prior experiments, and they suggest that: (1) self-efficacy and negative incentive value interact in their effects on blood pressure; (2) the cognitive regulation of systolic and diastolic blood pressure and pulse pressure can be distinguished; (3) the subjects with low self efficacy and high negative incentive value are the ones who generally experience the greatest activation; and (4) perceived competence does not appear to contribute to the psychophysiological regulation. PMID- 16360878 TI - Maternal personality and reproductive ambition in women is associated with salivary testosterone levels. AB - Previous research has linked testosterone levels with sex-specific personality traits within women. The present study investigates the relation between salivary testosterone levels and specifically maternal personality traits in healthy adult women. Twenty-seven young women completed the Bem Sex Role Inventory (BSRI). Additional questions were asked about maternal personality (importance of having children, self-rated maternal/broodiness), reproductive ambition (ideal number of children, ideal own age at first child) and career orientation (importance of having career). Higher circulating testosterone levels were associated with lower scores on measures of maternal personality and reproductive ambition. There was no relation of career orientation with testosterone. A median split on BSRI masculinity revealed high scorers had higher testosterone levels than low scorers. There was no relation of BSRI femininity with testosterone. Results suggest maternal tendencies may be partly androgen driven. PMID- 16360879 TI - Gender specific gene-environment interactions on laboratory-assessed aggression. AB - We examined gene-environment interactive effects on aggressive behavior among men and women genotyped (short versus long alleles) for the serotonin transporter gene. Aggressive behavior was indexed via a laboratory paradigm that measured the intensity and duration of shocks delivered to a putative "employee". Half of the participants were exposed to a physical stressor during the procedure (stress) and half were not (no-stress). Participants' physiological responses were gauged via acoustic startle eyeblink reactions (startle reactivity). Results were that men with the homozygous short (s/s) genotype showed increased aggression only under stress, whereas women and men carrying the long allele did not show differences in aggression in stress versus no-stress. However, although stress exposure produced increases in startle reactivity, there were no genotype or gender differences in physiology. These results replicate longitudinal research findings confirming the interactive effects of genes and environment on behavioral reactivity and on the development of externalizing psychopathological syndromes, at least in men. PMID- 16360880 TI - The relation of cortical activity and BIS/BAS on the trait level. AB - The behavioral activation system (BAS) and the behavioral inhibition system (BIS) have been proposed to be related to anterior asymmetry in the BIS/BAS model of anterior asymmetry. As an alternative, it may be suggested that behavioral activation comprises approach and withdrawal motivation and that the BAS is related to bilateral frontal trait activity. The aim of the present study was an empirical investigation on the relation between cortical trait activity, BIS and BAS. Data of 59 subjects on four occasions were analyzed. On each occasion, cortical activity was evaluated with resting EEG, and the BIS and BAS strength was assessed with questionnaires. Subjects with greater bilateral frontal cortical activity showed higher BAS scores. The latter result may suggest that behavioral activation comprises approach and withdrawal motivation. PMID- 16360881 TI - Comparison between mismatch negativity amplitude and magnetic mismatch field strength in normal adults. AB - The auditory mismatch negativity (MMN) or its magnetic counterpart (magnetic mismatch field, MMF) has been widely used to assess the ability of stimulus driven change detection process in humans. The authors evaluated the similarity of inter-individual variation of the response strength between MMN and MMF recordings. Three types of MMN or MMF were recorded in ten healthy subjects: change in duration of pure-tone stimuli, change in duration of the Japanese vowel /a/, and difference between the Japanese vowels /a/ and /o/. There was no significant correlation between MMN amplitude and MMF strength under any condition and in either hemisphere. These results suggest that widely used indices of MMN in the two technologies, i.e., EEG-amplitude and MEG-ECD may not be proportional in an individual. To further clarify the differential significance of recording MMN/MMF may be important to establish MMN/MMF as clinical indices of individual ability of preattentive stage of auditory processing. PMID- 16360882 TI - Combined effects of alcohol and caffeine on the late components of the event related potential and on reaction time. AB - The effects of .7 ml/kg alcohol and 200 mg caffeine on the P200, N200, P300 and N500 difference wave components of the event-related potential and on reaction time (RT) were examined in 16 females who performed both simple and choice RT tasks. Alcohol slowed the decision time (DT) component of reaction time, lengthened the latency of the P200 and P300 components, reduced N200 amplitude, increased P300 amplitude at parietal sites, and modified the effect of sagittal site on N500 difference wave peak amplitude. Caffeine shortened DT in the choice RT task, shortened N200 latency at right hemisphere sites, and shortened N200 latency in the choice RT task in combination with alcohol compared to when alcohol was administered alone. Caffeine also increased P300 amplitude in the choice RT task and reduced the integral of the N500 difference wave at most sites when combined with alcohol. It was concluded that whereas alcohol slows attention allocation and impairs working memory, caffeine accelerated response-related decisions and enhanced cortical arousal. PMID- 16360883 TI - Finger lengths, 2D:4D ratios, and their relation to gender-related personality traits and the Big Five. AB - Finger lengths and the ratio of index finger to ring finger length (2D:4D) may be markers of gonadal hormone exposure. The current study investigated possible associations between absolute finger lengths, 2D:4D ratios, and gender-related personality traits in over 2000 participants. Regression analyses showed no associations between men's 2D:4D ratios and gender-related personality traits and weak associations between women's 2D:4D ratios and gender-related occupational preferences. Men's absolute finger lengths were weakly associated with self ascribed masculinity, and women's absolute finger lengths were weakly associated with masculine occupational preferences. Big Five personality traits were assessed in a subsample of over 1000 participants. Analyses showed a weak positive association between 2D:4D and extraversion and a weak negative association between 2D:4D and openness to experience. Absolute finger lengths showed a weak negative association with agreeableness and a tendency to be associated with women's but not men's openness. Overall, associations between finger-length measures and personality were weak and inconsistent. PMID- 16360884 TI - The major histocompatibility complex class II transactivator is differentially regulated by interferon-gamma and transforming growth factor-beta in microglial cells. AB - We evaluated the regulation of the major histocompatibility complex class II (MHC II) transactivator (CIITA) gene expression in two microglial cell lines, EOC2 and EOC20. We demonstrate that interferon-gamma (IFN-gamma) activates type III- and IV-CIITA mRNA and high levels of MHC II in EOC20. However, in EOC2 cells only low levels of type IV-CIITA mRNA and MHC II are detectable following IFN-gamma treatment. Transforming growth factor-beta1 (TGF-beta1) inhibits both type III- and IV-CIITA expression in EOC20 cells while, in EOC2 cells TGF-beta1 enhances IFN-gamma induced pIV-CIITA expression. Trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, abrogates the TGF-beta1 mediated repression of the IFN-gamma induced CIITA in EOC20. Evidence is presented that the TG-interacting factor (TGIF), a co-repressor known to recruit HDACs, plays a role in determining the effects of TGF-beta1 on microglial cells. PMID- 16360885 TI - Multiple toll-like receptor agonists act as potent adjuvants in the induction of autoimmunity. AB - Infections can trigger or exacerbate the course of Multiple Sclerosis, and both bacterial and viral agents have been implicated. These agents are recognized by host cells via pathogen-associated molecular patterns activating TLRs. We investigated the role that PAMPs play in the animal model Experimental Autoimmune Encephalomyelitis, and found various MyD88-dependent PAMPs can participate as the adjuvant to induce EAE. Studies with IRAK1-deficient mice suggest that signaling through TLRs is not required in the target organ to develop disease. This suggests that PAMPs play an important role in priming of autoreactive T cells in EAE and potentially MS. PMID- 16360886 TI - CD25 regulatory T cells determine secondary but not primary remission in EAE: impact on long-term disease progression. AB - Multiple sclerosis (MS) is often characterized by several relapses and remissions during long-term disease, but neither the responsible cells nor the mechanisms are known to date. Using an animal model of multiple sclerosis, relapsing experimental autoimmune encephalomyelitis (R-EAE) CD4+CD25+ Treg cells expressing Foxp3 and CTLA-4 intracellularly and T lymphocytes expressing surface CTLA-4 were identified in the CNS. The first remission occurred even after depletion of Treg cells, but secondary remissions from EAE were ablated. Despite the unaltered first remission autoantigen rechallenge revealed already an amplified cytokine response during acute phase. These results indicate that the cellular composition during first attack of MS predicts long-term disease progression. PMID- 16360887 TI - Olfactory dysfunction: a sequela of pediatric blunt head trauma. AB - OBJECTIVE: To evaluate olfactory function in children with blunt head trauma. METHODS: Eligible subjects were consecutive children presenting at San Diego Children's Hospital Emergency Department or Trauma Service with blunt head trauma. Thirty-seven head injured children mean age 10.11+/-2.74 were evaluated; 36 healthy age and gender matched community children, mean age 10.08+/-2.99, served as controls. The medical record was reviewed for clinical measures related to head injury (HI). All children underwent olfactory assessment with the San Diego Children's Odor Identification Test (SDOIT). Olfactory event-related potentials were recorded on a subset of head injured and control children. RESULTS: Three of 37 head injured children had olfactory dysfunction. The head injured hyposmics had signs of more severe head injury represented by lower Glasgow Coma Scores (GCS), 10.33+/-5.51, when compared to their normosmic counterparts with mean GCS of 14.06+/-2.82. When stratifying the head injured children by clinical measures, lower Glasgow Coma Score, and abnormalities on head CT scan were associated with poorer olfactory performance. CONCLUSION: Children with blunt head injury may suffer post-traumatic olfactory impairment. More severe head injuries are more likely to produce olfactory deficits. In cases of significant head injury, cranial nerve I evaluation is imperative. PMID- 16360888 TI - Competition for sucrose pellets in tetrads of male Wistar, Fischer or Sprague Dawley rats: is intra-group ranking reflected in the level of anxiety? AB - Competition for palatable food or fluids within groups of rats has been previously used to mirror intra-group ranking. The paradigm of competition for sucrose pellets in non-food-deprived male Wistar rats was here extended from triads to tetrads aiming at evaluating whether the number of poor-performing rats, those animals being likely to model aspects of human psychopathologies (anxiety/depression/social withdrawal), could be increased. To evaluate potential superiority over the previously used Wistar strain, establishment and stability of the ranking was also assessed in tetrads of male Fischer and Sprague-Dawley rats. Clear and stable rank orders were seen in around 60-70% of both triads and tetrads of Wistar rats: a high-performing, a medium-performing and one (in triads) or two (in tetrads) poor-performing rats were identifiable, indicating that the number of poor-performing rats had increased in tetrads. Comparable rank orders were also seen in tetrads of Fischer and Sprague-Dawley rats. At the end of an extended period of repeated testing, tetrads of these two strains, as well as some selected Wistar tetrads, were tested in the elevated zero-maze and plasma corticosterone levels were determined. The differentiation in competition performance among cage mates was not paralleled by a difference in performance in the elevated zero-maze or in plasma corticosterone levels in any of the three strains. These data indicate that the level of anxiety in a non-social paradigm, the elevated zero-maze, does not reflect the competition-performance within the home cage and thus, the dominant/subordinate status in this food-competition paradigm may not reflect/being caused by different levels of anxiety. PMID- 16360889 TI - The effects of chronic administration of quetiapine on the phencyclidine-induced reference memory impairment and decrease of Bcl-XL/Bax ratio in the posterior cingulate cortex in rats. AB - Quetiapine, a new atypical antipsychotic drug, effectively alleviates positive and negative symptoms, as well as cognitive impairment that may be caused by neurodegeneration, in schizophrenia patients. Earlier in vivo and in vitro studies have demonstrated that quetiapine may be a neuroprotectant. The present study was designed to examine the beneficial effects of quetiapine on the possible cognitive impairment and changes of brain apoptotic regulation proteins induced by phencyclidine (PCP) in rats. Rats were treated with quetiapine (10 mg/kg/day; intraperitoneal (i.p.)) or vehicle for 16 days. On day 14, 1 h after the administration of quetiapine, the rats were given PCP (50 mg/kg; subcutaneous (s.c.)) or vehicle. Then quetiapine was administrated for an additional 2 days. One day after the last quetiapine injection (3 days after the PCP injection), the rats were trained on a spatial memory task in a radial arm maze. After the behavioural test, the rats were decapitated for Western blot analysis. PCP induced reference memory impairment, and a decrease of the ratio of an anti apoptotic Bcl-2 family member (Bcl-XL) to a pro-apoptotic analogue (Bax) in the posterior cingulate cortex. Chronic administration of quetiapine counteracted the PCP-induced reference memory impairment and decrease of Bcl-XL/Bax ratio in the posterior cingulate cortex. These results suggest that quetiapine may have ameliorating effects on the cognitive impairment and brain apoptotic processes induced by PCP. PMID- 16360892 TI - Modulation of ERK1/2 and p38(MAPK) by lead in the cerebellum of Brazilian catfish Rhamdia quelen. AB - Lead (Pb2+) is a neurotoxic trace metal, widespread in aquatic environment that can change physiologic, biochemical and behavioral parameters in diverse fish species. Chemical exposure may drive modulation of mitogen-activated protein kinases (MAPKs) that are a family of highly conserved enzymes which comprise ubiquitous groups of signaling proteins playing critical regulatory roles in cell physiology. Extracellular signal-regulated kinases (ERK1/2) and p38(MAPK) control complex programs such as gene expression, embryogenesis, cell differentiation, cell proliferation, cell death and synaptic plasticity. Little information is available about MAPKs in aquatic organisms and their modulation by trace metals. The aim of this work was to determine the modulation of ERK1/2 and p38(MAPK) phosphorylation by Pb2+ in vivo and in vitro, in cerebellar slices of the catfish, Rhamdia quelen. In the in vitro model, slices were incubated for 3 h with lead acetate (1-10 microM). In the in vivo studies, the animals were exposed for 2 days to lead acetate (1 mg L(-1)). ERK1/2 and p38(MAPK) (total and phosphorylated forms) were immunodetected in cerebellar slices by Western blotting. Pb2+ added in vitro at 5 and 10 microM increased significantly the phosphorylation of both MAPKs. The in vivo exposed animals also showed a significant increase of ERK1/2 and p38(MAPK) phosphorylation without changes in the total content of the enzymes. In conclusion, the present work indicates that it is possible to evaluate the ERK1/2 and p38(MAPK) activation in the central nervous system (CNS) of a freshwater fish largely distributed in South America. Moreover, Pb2+, an important environmental pollutant may activate in vitro and in vivo ERK1/2 and p38(MAPK) enzymes. These findings are important considering the functional and ecologic implications associated to Pb2+ exposure of a freshwater fish species, such as R. quelen, and the roles of ERK1/2 and p38(MAPK) in the control of brain development, neuroplasticity and cell death. PMID- 16360893 TI - Rapid detection and species identification of Mycobacterium spp. using real-time PCR and DNA-microarray. AB - Infections with mycobacteria are an important issue in public health care. Here we present a "proof-of-principle" concept for the identification of 37 different Mycobacterium species using 5' exonuclease real-time PCR and DNA microarray based on the region upstream of the 65 kDa heat shock protein. With our two PCR probes, one complementary to all mycobacteria species, the other specific for the M. tbc complex, 34 species were properly classified by real-time PCR. After reamplification and hybridization to a DNA microarray, all species showed a specific pattern. All 10 blindly tested positive cultures revealed a positive real-time PCR signal with the genus probe. After reamplification and hybridization, six samples could unambiguously be identified. One sample showed a mixture of presumably three species-specific patterns and sequencing the 16S rRNA confirmed the presence of a mixture. The hybridization results of three specimens could not be interpreted because the signal to background ratio was not sufficient. Two samples considered as negative controls (LAL Reagent Water (Cambrex) and DNA of Candida albicans) gave neither a genus nor a M. tbc-complex positive PCR signal. Based on these results we consider our method to be a promising tool for the rapid identification of different mycobacteria species, with the advantage of possible identification of mixed infections or contaminations. PMID- 16360894 TI - Normal clotting. AB - OBJECTIVE: To review the normal coagulation process and the mechanisms that lead to abnormal clotting. DATA SOURCES: Primary and tertiary literature and the authors' clinical experience. CONCLUSION: The process of coagulation is complex and can be easily misunderstood. It is important to be familiar with normal coagulation before one can comprehend the coagulopathies associated with malignancies. IMPLICATIONS FOR NURSING PRACTICE: A thorough understanding of the coagulation process is a critical prerequisite to caring for patients with clotting disorders. Once the normal clotting process is understood, the abnormal becomes easier to recognize and the cancer-associated dysfunctions more readily identified. PMID- 16360895 TI - Abnormal clotting in cancer: an overview of pathophysiology and etiology. AB - OBJECTIVE: To help oncology nurses become more cognizant of the increased risk cancer patients have for venous thromboembolism due to such factors as chemotherapy and hypercoagulability stemming from the neoplasm itself. DATA SOURCES: Primary and tertiary literature and the author's clinical experience. CONCLUSION: Establishing the optimal treatment of cancer-associated thrombosis in the future will depend in part on a better understanding of clotting aberrations. Oncology nurses must be cognizant of the various risk factors for thrombosis and bleeding and how they relate to cancer treatments and interventions. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses are in a unique position to recognize conditions that predispose cancer patients to the development of venous thromboembolism as well as its signs and symptoms. This enhanced understanding will ensure that patients receive appropriate and timely intervention. PMID- 16360896 TI - Therapeutic options for the treatment of cancer-associated thrombosis. AB - OBJECTIVE: To provide oncology nurses with an understanding of therapeutic options for cancer-associated thrombosis, strategies to prevent recurrence, and practical issues in patient management. DATA SOURCES: Primary and tertiary literature and the author's clinical experience. CONCLUSION: Oncology nurses monitor patients throughout the care continuum for signs and symptoms indicating vascular thromboembolism and need to know the steps to take to expedite an accurate diagnosis and ensure prompt treatment. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses must keep informed about the evolving evidence that leads to practice changes. They should be able to teach patients about therapeutic options and their potential for improving outcomes. PMID- 16360897 TI - Use of low-molecular-weight heparins in the treatment and secondary prevention of cancer-associated thrombosis. AB - OBJECTIVE: To provide oncology nurses with an understanding of the differences between low-molecular-weight heparins and unfractionated heparin, key clinical trial data, and current treatment guidelines. DATA SOURCES: Primary and tertiary literature and the authors' clinical experience. CONCLUSION: The latest American College of Chest Physicians guidelines recommend low-molecular weight heparin for at least the first 3 to 6 months of long-term treatment for most patients with cancer and deep vein thrombosis (DVT; grade 1A). IMPLICATIONS FOR NURSING PRACTICE: Anticoagulant therapy has been found to be the most effective form of treatment for venous thromboembolism in patients with cancer, and oncology nurses play a critical role in the choice of agents, the dispensing of drugs, and the monitoring of ongoing therapy. Vital to that role are an ability to differentiate among the available agents and apply the current practice guidelines based on the clinical trial data. PMID- 16360898 TI - Nursing interventions in the management of patients with cancer-associated thrombosis. AB - OBJECTIVES: To provide oncology nurses with an overview of the clinical trial evidence and to help them apply the data to patient management to optimize care and ensure the best possible outcomes. DATA SOURCES: Primary and tertiary literature and the authors' clinical experience. CONCLUSION: Nursing interventions for patients with cancer-associated thrombosis involve such key areas as the need for careful and continuing risk assessment, close monitoring of laboratory values, evaluation of patients as candidates for outpatient management, patient and caregiver education, and monitoring of patient compliance. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses provide critical oversight and coordination in the treatment and prevention of venous thromboembolism. These interventions are essential to reducing venous thromboembolism-associated morbidity and mortality. PMID- 16360899 TI - Combined analysis of association between personality traits and three functional polymorphisms in the tyrosine hydroxylase, monoamine oxidase A, and catechol-O methyltransferase genes. AB - Several molecular genetic studies have been conducted with regard to the association between catecholamine-related genes and personality traits. However, the results of replication studies did not always coincide. One of the possible reasons may be that the effect exerted by the individual gene is small. In the present study, we investigated the association between personality traits and systematic combination of functional polymorphisms in three genes that regulate the metabolism of catecholamines, namely, tyrosine hydroxylase (TH), monoamine oxidase A (MAOA), and catechol-O-methyltransferase (COMT). The (TCAT)n repeat in the TH gene, the promoter variable number tandem repeat (VNTR) in the MAOA gene, and Val158Met in the COMT gene were genotyped in 256 healthy Japanese volunteers. Personality traits were evaluated using the NEO Personality Inventory-Revised (NEO PI-R). As a result, the score for Neuroticism increased, and those for Extraversion and Conscientiousness decreased according to the degree of functional polymorphic change, i.e., the lower synthesis/higher catalysis of catecholamines. A statistically significant difference was observed in the change of Extraversion (p=0.04, after Bonferroni correction). These results may provide evidence for the association between metabolic change of catecholamines and personality traits, which may be due to the additive effect of the three genes. PMID- 16360900 TI - Gene expression analysis using a modified HiCEP method applicable to prokaryotes: a study of the response of Rhodococcus to isoniazid and ethambutol. AB - We established a novel method to analyze the gene expression of prokaryotes by modifying and optimizing the HiCEP (high coverage gene expression analysis) method, which was originally developed for eukaryotic gene expression profiling. Following the removal of abundant rRNA, the mRNA of prokaryotic cells was enzymatically polyadenylated and subjected to HiCEP analysis. This method was highly reproducible due to selective PCR that was performed by using adaptor specific primers. We confirmed induction of tipA and induction or suppression of cspA, which are genes that are obtained from distinctive actinomycetes and responded to thiostrepton and temperature stress, respectively. Further, we applied this method to explore the gene expression profile of Rhodococcus erythropolis in response to drugs that inhibit cell wall synthetic pathways, and we were able to identify 35 upregulated genes. Among these genes, we confirmed the upregulation of 22 genes by using RT-PCR (reverse transcriptase-polymerase chain reaction). Some of these genes are involved in the synthesis of mycolic acid and arabinogalactan, suggesting a cell response to drug treatment by regulation of the genes involved in cell wall synthesis. This method could prove to be a useful technique for gene expression analysis of prokaryotes, particularly nonmodel strains with unknown genome sequences. PMID- 16360902 TI - CONSORT and QUOROM guidelines for reporting randomized clinical trials and systematic reviews. PMID- 16360905 TI - Stability and functional appliances. PMID- 16360906 TI - When are implants needed? PMID- 16360907 TI - A comparison of providers' and consumers' perceptions of facial-profile attractiveness. AB - INTRODUCTION: The purpose of this study was to evaluate the perception of facial attractiveness in profile digital photographs that were incrementally altered to produce different combinations of mandibular anteroposterior positions and lower anterior facial heights. The specific aims were to determine whether there is concordance between providers and consumers in their perceptions of facial attractiveness, and to evaluate whether interactions of the anteroposterior and vertical dimensions and the magnitude of these interactions influence perception of facial attractiveness. METHODS: Profile digital photographs and cephalograms of 3 men and 3 women were used. The position of the jaw was altered incrementally with Dolphin imaging equipment (Dolphin Imaging and Management, Chatsworth, Calif), and booklets were created. One hundred raters (50 laypersons, 25 oral surgeons, 25 orthodontists) scored the profiles on a visual analog scale. All images were duplicated to test intrarater reliability. Factorial ANOVA with repeated measures and the Tukey-Kramer post-hoc test for multiple comparisons were used to test for differences in facial attractiveness. The level of significance was set at .05 (P = .05) for all analyses. RESULTS: Intrarater reliability was good (ICC = 0.71), and general concordance was found between providers and consumers in their perceptions of facial attractiveness. Interactions of the anteroposterior and vertical dimensions and the magnitude of change in each dimension influence the perception of facial attractiveness. CONCLUSIONS: The results suggest that preferences of facial attractiveness by laypersons, orthodontists, and oral surgeons in central Ohio are generally in agreement. This information might assist clinicians in treatment planning and making recommendations. PMID- 16360908 TI - Patient- and clinician-perceived need for orthognathic surgery. AB - INTRODUCTION: To better understand why patients believe they need orthognathic treatment, a study was undertaken to examine perceived need for treatment by patients and clinicians. This questionnaire-based study was undertaken at The John Radcliffe Hospital, Oxford, United Kingdom. SUBJECTS AND METHODS: Forty patients were recruited from combined orthodontic-surgical clinics. They were asked to rate their perceived need for treatment based on facial appearance, dental appearance, function, and overall need. Twenty orthodontists and 20 maxillofacial surgeons were asked to rate perceived need for treatment based on the same parameters, using study models and clinical photographs. Ratings were marked on visual analog scales. RESULTS: Significant differences were found between patients and clinicians in perceived need for treatment based on facial appearance (orthodontists compared with patients, P = .023; surgeons compared with patients, P = .001). In addition, maxillofacial surgeons rated a significantly greater overall need for treatment than patients (P = .027), and they rated treatment need based on facial appearance (P = .005) and function (P < .001) significantly higher than orthodontists. CONCLUSIONS: Clinicians rated greater need for orthognathic treatment based on facial appearance than did patients. Surgeons also rated greater overall need for treatment than patients. In addition, surgeons rated treatment need based on facial appearance and function significantly higher than orthodontists, but large variations existed in both clinician groups. PMID- 16360909 TI - Index of orthodontic treatment need as a predictor of orthodontic treatment uptake. AB - INTRODUCTION: The aim of this study was to evaluate whether the index of orthodontic treatment need (IOTN) could be weighted by using consumer-based sociodental measures to predict the uptake of orthodontic services. METHODS: The sample consisted of 525 schoolchildren, 11 to 12 years old, from Greater Manchester, United Kingdom. Child-perceived IOTN aesthetic component (AC), examiner IOTN AC and dental health component, and child socioeconomic status (Townsend score) were recorded. Two consumer sociodental measures (utility and oral aesthetic subjective impact scale values) were recorded. Three years later, the proportion of the subjects who had received orthodontic treatment or were on a waiting list for orthodontic treatment was recorded, and the rate of service uptake was determined. RESULTS: Sociodental indicators did not predict uptake of orthodontic services. A child with higher normative clinical treatment need was 3 times more likely to receive orthodontic treatment than a child with low clinical need (P < .05). CONCLUSIONS: Consumer-based sociodental information does not predict future use of orthodontic services. Factors such as clinical IOTN and child-perceived IOTN AC will adequately predict use of orthodontic services. PMID- 16360910 TI - Eligibility for publicly funded orthodontic treatment determined by the handicapping labiolingual deviation index. AB - INTRODUCTION: Access to orthodontic care for Medicaid patients has been limited, in part because of orthodontists' reluctance to treat severe malocclusions for low reimbursements. Limited orthodontic treatment in the mixed dentition (phase 1 treatment) has been proposed to address this issue, because the intent of phase 1 treatment is to improve or prevent severe malocclusions. Orthodontists might be more willing to provide shorter, simpler treatment. The purpose of this study was to determine whether phase 1 treatment would reduce malocclusion severity to the extent that eligibility for subsequent Medicaid-funded treatment was significantly reduced. METHODS: Eligibility was determined by the handicapping labiolingual deviation (HLD) index, which is used by several states for this purpose. Eligibility was also determined with the index of complexity, outcome, and need (ICON). This allowed us to compare these 2 indexes. Pre-phase 1 and post phase 1 index scores were calculated by using study casts from 193 patients treated at the University of Washington orthodontic clinic and the Odessa Brown Children's Dental Clinic, both in Seattle. RESULTS: Using the HLD index, we found that eligibility for orthodontic treatment decreased by 62% after phase 1 treatment. This change was statistically significant at P < .0001. The ICON found significantly more treatment need before phase 1 (90%) than did the HLD index (35%) (P < .0001). CONCLUSIONS: Early interceptive treatment significantly reduces eligibility for comprehensive Medicaid-funded orthodontic treatment. The HLD index is a useful tool for determining Medicaid eligibility. PMID- 16360911 TI - Transverse growth of the maxilla and mandible in untreated girls with low, average, and high MP-SN angles: a longitudinal study. AB - INTRODUCTION: The purpose of this study was to investigate maxillary and mandibular transverse growth in untreated female subjects with low, average, and high mandibular plane angles longitudinally from ages 6 to 18. METHODS: Eighty one untreated white girls with low (< or = 27 degrees , n = 16), average (> 27 degrees to < 37 degrees , n = 41), and high (> or = 37 degrees , n = 24) mandibular plane angles at age 6 were selected from the Bolton-Brush and Burlington Growth Studies. For each subject, longitudinal posteroanterior cephalograms at different ages (from ages 6 to 18) were traced, and the widths of maxilla and mandible were measured. All the measurements were converted by using a magnification factor of 8.5% (the subject-to-film distance was set at 13 cm). RESULTS: At age 6, the high-angle group had narrower maxillary and mandibular widths than the low-angle group, and this trend continued until age 18. From ages 6 to 14, maxillary width showed a steady and similar rate of increase for all 3 groups (0.90-0.95 mm per year), yet a plateau was reached at age 14 for all groups. Mandibular width increased at a steady rate (about 1.6 mm/year) for all 3 groups until age 14, and a plateau was reached for the high-angle group. For the low- and average-angle groups, mandibular growth continued from ages 14 to 18 but at a slower rate (0.85 mm and 0.39 mm per year, respectively). CONCLUSIONS: Vertical facial patterns (with low or high mandibular plane angles) might play a strong role in the transverse growth of the maxilla and the mandible. PMID- 16360912 TI - Submentovertex cephalometric norms in Turkish adults. AB - INTRODUCTION: The aims of this study were (1) to establish cephalometric norms from submentovertex (SMV) radiographs for young Anatolian Turkish adults and (2) to identify possible sex differences between men and women. METHODS: SMV cephalograms were taken of 50 nongrowing Turkish adults (25 men, mean age, 23.06 +/- 2.30 years; 25 women, mean age, 22.10 +/- 2.04 years). All had normal occlusions and well-balanced faces. Five angular and 10 linear measurements were made on each radiograph. For each variable, arithmetic mean and standard deviation minimum and maximum values were calculated. Independent-sample t tests were performed for the sex comparisons. Cephalometric angular and linear norms for Turkish adults were determined by using the Lew-Tay SMV cephalometric analysis. SMV cephalometric standards for Turkish adults were also developed. RESULTS: Generally, our measurements were similar to the Lew-Tay SMV norms. Most Turkish SMV cephalometric measurements showed statistically significant sex differences. Comparison between Turkish men and women indicated larger measurements for men in all investigated linear measurements. CONCLUSIONS: SMV cephalometric norms are useful in the diagnosis of mandibular asymmetries and the treatment of dentofacial orthopedics in Turkish patients. PMID- 16360914 TI - Effect of argon laser curing on the shear bond strength of metal brackets bonded with light-cured glass ionomer cement. AB - INTRODUCTION: The purpose of this study was to evaluate the shear bond strength and debonding characteristics of glass ionomer cement cured with an argon laser. METHODS: Thirty extracted first premolars were divided into 2 groups of 15. The teeth were cleaned and mounted in resin, and metal brackets were bonded with glass ionomer cement. In the control group, the bond was cured with a halogen light for 40 seconds (20 seconds mesial and 20 seconds distal). In the test group, the specimens were cured with an argon laser for 5 seconds. Brackets were debonded in shear; bond strength was measured, and the adhesive remnant index was scored. RESULTS: The difference in mean shear bond strength was not statistically significant between groups when compared with a t test. More adhesive remained on the enamel surface in the laser group than in the control group. CONCLUSIONS: Argon laser curing produces bond strengths equivalent to those obtained with traditional light curing in much less time, but it leaves more adhesive on the tooth surface. PMID- 16360913 TI - Craniofacial and dentofacial development in pigs fed soft and hard diets. AB - INTRODUCTION: Several authors have found a correlation between reduced chewing activity and malocclusion. In animal experiments, it has been possible to correlate a diet of low chewing resistance to narrower arches, which predispose the subject to crowding and irregular teeth. METHODS: In this study, 17 pigs were weaned at 5 weeks of age and divided into 2 groups according to diet and housing. The soft-diet, indoor group was housed in conventional pens, and the animals were fed aliquots of barley and oats with soya and a mineral-vitamin premix added. The food was mixed with water at a dry matter content of about 25%. The hard-diet, outdoor group was kept outdoors; the pigs were fed solid food and also ate organic matter in the soil. All pigs were killed at 22 months of age. The skulls were dissected, and transverse and sagittal craniofacial and dentofacial dimensions were measured. RESULTS: Chewing hard food caused considerable occlusal and approximal attrition in the experimental animals. The dental arches were shorter due to this attrition and to mesial migration of the molars and premolars. The approximal attrition also reduced the tendency for crowding and rotation of the teeth. Posterior crossbite was more common among the hard-chewing animals. CONCLUSIONS: The increase in arch width in the hard-diet animals reported in the literature was not duplicated in this study. On the contrary, the soft-diet pigs had significantly wider arches than the hard-diet animals. This was most pronounced in the premolar region and could be the result of an atypical tongue habit, caused by the nonphysiologic feeding of the pigs. The soft-diet pigs also had a greater tendency to postnormal occlusion, especially in the canine region. PMID- 16360915 TI - Effect of shorter polymerization times when using the latest generation of light emitting diodes. AB - INTRODUCTION: Recent studies have suggested that a 10-second cure time with a high-energy quartz-tungsten-halogen (QTH) or a light-emitting diode (LED) light might be adequate when bonding orthodontic brackets to tooth enamel. The purpose of this study was to evaluate the ability of the latest generation of QTH and LED light-curing units (LCUs) to bond orthodontic brackets to teeth at decreased polymerization times. METHODS: Two LED LCUs (Ortholux LED, 3M Unitek, Monrovia, Calif; UltraLume LED 5, Ultradent Products, South Jordan, Utah) and a QTH LCU (Optilux 501, Demetron, Danbury, Conn) were evaluated. One hundred eighty metal orthodontic brackets were bonded to extracted human molars. The specimens were divided into 9 groups (3 lights and 3 curing times) of 20 teeth each. Each group was cured with 1 of the 3 lights for 20, 10, or 6 seconds. Thirty minutes after polymerization, the specimens were subjected to shear force on a universal testing machine until bracket failure. RESULTS: Two-way ANOVA detected significant differences among the main effects of light type and cure time. Tukey post-hoc tests determined that brackets bonded by all light types had lower bond strengths with the 6-second cure than the 20-second cure (P < .001). The highest bond strengths were obtained with the Optilux 501 QTH LCU and the UltraLume LED 5 LCU at the longest cure time of 20 seconds. CONCLUSIONS: It is recommended that orthodontic brackets be photopolymerized for at least 20 seconds with the QTH or the LED LCU before the archwires are engaged. PMID- 16360916 TI - Light-curing time reduction with a new high-power halogen lamp. AB - INTRODUCTION: Orthodontic brackets are routinely bonded with light-cured adhesives. Conventional halogen lights used in bonding have the disadvantage of a long curing time, and the available alternatives (laser and plasma lights) are expensive. Our aim was to investigate the minimum time necessary to bond brackets with a new, relatively low-priced, high-power halogen light. METHODS: Five groups of 15 deciduous bovine incisors were bonded with stainless steel brackets (Mini Diamond Twin, Ormco, Orange, Calif) by using different lamps and curing times. Three of the groups were bonded by using a high-power halogen light (Swiss Master Light, Electro Medical Systems, Nyon, Switzerland) for 2, 3, and 6 seconds, respectively. The fourth group, bonded with a fast halogen light (Optilux 501, Sybron Dental Specialties, Danbury, Conn) for 40 seconds, served as the positive control group. The fifth group, the comparison group, was bonded with a plasma light (Remecure, Remedent, Deurle, Belgium) for 4 seconds. After storage for 24 hours in the dark at 37 degrees C in water, shear bond strength was measured with a universal testing machine. RESULTS: A curing time of 2 seconds with the high power halogen light negatively affected the bond strength and the probability of bond survival. The adhesive remnant index scores were not significantly different among the groups. Most failures (> 60%) occurred at the bracket base/adhesive interface. CONCLUSIONS: The high-power halogen light seems to be a cost-effective solution to reducing curing time. The recommended curing times to bond stainless steel brackets are 6 seconds and, with caution, even 3 seconds. PMID- 16360917 TI - The effect of tooth bleaching on the shear bond strength of orthodontic brackets. AB - INTRODUCTION: The purpose of this study was to determine the effect of enamel bleaching on the shear bond strength of orthodontic brackets bonded with a composite adhesive. METHODS: Two protocols were used on 60 human molars. In the at-home bleaching group (n = 30), Opalescence bleaching agent (Ultradent, South Jordan, Utah), which contains 10% carbamide peroxide, was brushed onto the teeth daily for 14 days and left for 6 hours each day. Teeth in the in-office group (n = 30) were treated with Zoom! (Discus Dental, Culver City, Calif), which contains 25% hydrogen peroxide gel, and then exposed to a light source for 20 minutes; these teeth were treated twice. After bleaching, the specimens were randomly divided into equal subgroups and stored in artificial saliva at 37 degrees C for 7 or 14 days before bonding. Shear bond strength testing was performed on all teeth. The Kruskal-Wallis test for nonparametric means was used to determine whether significant differences existed between the various subgroups and an unbleached control group. RESULTS: The mean shear bond strength for the control group was 5.6 +/- 1.8 MPa. Means for the at-home groups were 5.2 +/- 3.6 MPa and 7.2 +/- 3.2 MPa for the 7- and 14-day waiting periods, respectively. Means for the in-office groups were 5.1 +/- 5.3 MPa and 6.6 +/- 2.6 MPa for the 7- and 14 day waiting periods, respectively. The Kruskal-Wallis test (X(2) = 8.089) indicated no significant differences between the 5 subgroups (P = .088). CONCLUSIONS: The results showed that in-office and at-home bleaching did not affect the shear bond strength of orthodontic brackets to enamel. PMID- 16360918 TI - Bond strength and debonding characteristics of a new ceramic bracket. AB - INTRODUCTION: The purposes of this study were to evaluate the shear bond strength of a new collapsible monocrystalline bracket (Inspire, Ormco, Orange, Calif) and compare it with another collapsible ceramic bracket (Clarity, 3M Unitek, Monrovia, Calif) and a metal bracket; to examine the modes of failure after shear bond strength testing; and to observe the tooth surface after debonding the ceramic brackets with pliers. METHODS: One hundred extracted human premolars were selected for bonding. Three types of brackets and 2 orthodontic adhesives (Transbond XT, 3M Unitek; and Enlight, Ormco) were used. After bonding, all samples were placed in a distilled water bath at 37 degrees C for 24 hours. The shear bond strength of 60 samples was measured, and the remaining 40 samples with ceramic brackets were debonded with pliers. RESULTS: No statistically significant differences in bond strengths among the different combinations of brackets and adhesives were found (P > .05). The mode of failure after debonding by either shear bond strength testing or with pliers was predominantly at the bracket/adhesive interface in all groups. Enamel and bracket fractures were noted in 2 and 5 of 20 samples for Inspire, and 1 and 0 of 20 samples for Clarity after debonding with pliers. CONCLUSIONS: Bond strength and mode of failure of Inspire were similar to those of Clarity, but the risk of bracket fracture after debonding for Inspire was greater. PMID- 16360919 TI - Relationship between masticatory function and internal structure of the mandible based on computed tomography findings. AB - INTRODUCTION: The purposes of this study were to investigate bone mineral density as a part of bone construction in human skulls and to examine the relationship between dentofacial morphology and masticatory function by using computed tomography (CT) findings. Changes in bone mineral density in the mandible because of loss of masticatory function were tested in rats by experimentally producing an environment that inhibited mastication by the molars. METHODS: Data for the human study were obtained from 27 modern male Japanese skulls (mean age, 28 years) from the University of Tokyo. Cortical bone thickness (CBT) and CT value (CV) were measured by each CT scan of the first and molars. For the animal study, a metal cap was inserted between the maxillary and mandibular incisors to prevent the molars from biting in 6-week-old male Wistar rats. The rats were killed after 2, 4, or 6 weeks, and bone mineral density was measured in cancellous and cortical bone equivalent to the first molar region by using peripheral quantitative computed tomography. RESULTS: In the human skull study, significant negative correlations were observed between CV in the regions of the buccal side of the second molar and the angle between the Frankfort horizontal and mandibular planes. Significant negative correlations were also observed between the gonial angle and CV in the buccal and basal sides. In the animal study, cancellous bone mineral density began to decline 4 weeks after the start of the experiment in the masticatory hypofunction group compared with the control group. By week 6, cancellous bone density had declined by 11.6% on the buccal side, 16.7% on the lingual side, 12.3% at the bifurcation of the root, and 38.1% at the root apex. Cortical bone density declined by 8% to 12% on the lingual side. CONCLUSIONS: The results support our hypothesis that a functional adaptive response by the mandible to mechanical stress resulting from mastication occurs not only in the muscle insertion area, but also in mandibular alveolar bone in the molar region. PMID- 16360920 TI - The mandibular muscles and their importance in orthodontics: a contemporary review. AB - It is widely accepted that the orthodontist should have a thorough understanding of the craniofacial musculature and its association with the growth and development of the dentofacial complex. There is still much controversy regarding the influence of the mandibular muscles on normal growth and development, and on orthodontic treatment and stability. This review presents an outline of the mandibular muscles and the vertical facial pattern. The different methods by which the mandibular muscles have been investigated are discussed. The potential influence of these muscles on normal morphologic variation in different people is also discussed, along with the implications for contemporary orthodontic treatment and stability. PMID- 16360921 TI - Noncompliant treatment of skeletal open bite. AB - The purpose of this article is to illustrate the effects of the rapid molar intrusion appliance, a treatment alternative that does not require patient compliance, for counteracting excessive vertical dimensions in growing patients and adults. The rapid molar intrusion appliance has 2 elastic modules that are secured on the maxillary and mandibular first-molar tubes. It is used in combination with maxillary and mandibular soldered lingual arches. Patient acceptance is good, and the patient's only responsibilities are to keep the appliance clean and avoid breakage. PMID- 16360922 TI - Extreme dentoalveolar compensation in the treatment of Class III malocclusion. AB - The orthodontic treatment of an adult patient with a skeletal Class III malocclusion, increased anterior facial height, negative overjet, and bilateral posterior crossbite is presented. Treatment options included mandibular first premolar or third molar extractions with dentoalveolar compensation or combined surgical-orthodontic treatment. Mandibular third molar extraction with dentoalveolar compensation was the treatment choice. Biofunctional brackets, with accentuated lingual crown torque on the maxillary incisors and accentuated buccal crown torque on the mandibular incisors, were used. The anterior crossbite was corrected with intermaxillary elastics from the palatal aspect of the maxillary incisors to the labial aspect of the mandibular incisors. Class III elastics moved the maxillary teeth mesially and assisted in retruding the mandibular teeth. Patient compliance with the elastics was excellent, and satisfactory dentofacial esthetics were achieved. This treatment protocol has rigorous indications, and it is not a routine plan. The mechanotherapy and the pros and cons of this approach are discussed. PMID- 16360923 TI - Orthodontics in 3 millennia. Chapter 6: More early 20th-century appliances and the extraction controversy. AB - The trying conditions of the Great Depression and World War II did not deter innovative orthodontists from adding 3 new appliances to our armamentarium. Clinicians become fragmented into various technique "camps." Silas Kloehn's neck gear became a more patient-friendly version of extraoral anchorage, but it still had drawbacks. Angle's stranglehold on the specialty was finally broken when 3 of his disciples made extractions respectable. PMID- 16360924 TI - Accuracy of linear temporomandibular joint measurements with cone beam computed tomography and digital cephalometric radiography. AB - INTRODUCTION: Cone beam computed tomography (CBCT) is making headway into imaging for orthodontics. The purpose of this study was to define CBCT multi-planar reformatted projections for temporomandibular joint (TMJ) examination and compare the accuracy of linear measurements of the TMJ and related structures from these projections with similar measurements made with conventional cephalograms and with the anatomic truth. METHODS: Linear dimensions between 11 anatomical sites were measured with a digital caliper to assess the anatomic truth for 25 dry human skulls. The skulls were imaged with iCAT (Xoran Technologies, Ann Arbor, Mich/Imaging Sciences International, Hatfield, Pa) CBCT, and cephalograms were made in all 3 orthogonal planes (lateral cephalometric [LC], posteroanterior [PA], and submentovertex [SMV]) acquired with photostimulable phosphor plates. Linear measurements were made on 7 custom CBCT reconstructions and the digital cephalograms. Modality means and the natural log of the standard deviations were compared post hoc against the actual dimensions by using analysis of variance with the Dunnett t test. Significance was set at P < .05. RESULTS: All CBCT measurements were accurate; however, 3 of 5 LC measurements, 4 of 5 PA measurements, and 4 of 6 SMV measurements varied significantly from the truth. Intraobserver CBCT measurements were highly reliable compared with anatomic truth and significantly more reliable than measurements made from LC, PA, and SMV images. CONCLUSIONS: Custom oblique multi-planar reformatted reconstructions with iCAT CBCT provide accurate and reliable linear measurements of mandibular and TMJ dimensions. PMID- 16360926 TI - Improvement of accessory symptoms of hypertension by TSUMURA Orengedokuto Extract, a four herbal drugs containing Kampo-Medicine Granules for ethical use: a double-blind, placebo-controlled study. AB - A double-blind, placebo-controlled study was conducted to evaluate the efficacy, safety, and utility of TSUMURA Orengedokuto Extract Granules for Ethical Use (TJ 15) as a treatment for the accessory symptoms of hypertension. Two capsules of the study drug were administered orally 3 times daily (i.e., before meals) for 8 weeks. Among 265 patients enrolled in the study, 134 were assigned to the TJ-15 group and 131 were assigned to the placebo group, of whom 204 patients (103 in the TJ-15 group and 101 in the placebo group) were included in the efficacy and utility analyze and 251 patients (128 in the TJ-15 group and 123 in the placebo group) were included in the safety analysis. Efficacy was significantly higher in the TJ-15 group based on the total score for the accessory symptoms of hypertensions which was the primary efficacy endpoint (Wilcoxon's rank sum test, p=0.013). When each accessory symptom of hypertension was assessed separately, efficacy was higher for hot flushes and facial suffusion in the TJ-15 group (Wilcoxon's rank sum test, p=0.034, and 0.022, respectively). There were no significant differences between the TJ-15 and the placebo groups with respect to the decrease of blood pressure or the antihypertensive effect. There was also no significant difference between the two groups with regard to the overall safety rating. The utility rating was significantly higher in the TJ-15 group than in the placebo group (Wilcoxon's rank sum test, p=0.016). In conclusion, TJ-15 was superior to placebo with respect to efficacy, safety, and utility for the treatment of accessory symptoms of hypertension. PMID- 16360925 TI - Cap-independent translation of plant viral RNAs. AB - The RNAs of many plant viruses lack a 5' cap and must be translated by a cap independent mechanism. Here, we discuss the remarkably diverse cap-independent translation elements that have been identified in members of the Potyviridae, Luteoviridae, and Tombusviridae families, and genus Tobamovirus. Many other plant viruses have uncapped RNAs but their translation control elements are uncharacterized. Cap-independent translation elements of plant viruses differ strikingly from those of animal viruses: they are smaller (<200 nt), some are located in the 3' untranslated region, some require ribosome scanning from the 5' end of the mRNA, and the 5' UTR elements are much less structured than those of animal viruses. We discuss how these elements may interact with host translation factors, and speculate on their mechanism of action and their roles in the virus replication cycle. Much remains to be learned about how these elements enable plant viruses to usurp the host translational machinery. PMID- 16360927 TI - Efficacy and tolerability of potato juice in dyspeptic patients: a pilot study. AB - In Europe, use of potatoes (Solani tuberosi tuberecens) is a traditional remedy for stomach complaints. We performed a pilot study on the effectiveness and tolerability of freshly squeezed potato juice in patients suffering from dyspeptic symptoms. After informed written consent, 44 patients with various dyspeptic symptoms were enrolled, to drink for 1 week twice daily 100ml or more of potato juice (Biotta, if complaints persisted, a further 100ml was recommended. Validated outcome measures included the gastrointestinal symptom (GIS) profile, a disease-specific health assessment questionnaire (QOLRAD) and self-rated treatment success (efficacy, a 5-point Likert Scale). The study was completed by 42 patients. The GIS and QOLRAD scores improved significantly by 41.9+/-40.8% (p=0.001) and 50.7+/-36.1% (p<0.001), respectively (ITT); the absolute values changed from 11.5+/-5.1 to 6.3+/-5.3 (GIS) and 62.0+/-25.7 to 32.0+/-28.8 (QORAD). Sixteen, 18 and 26 patients, respectively, rated the effectiveness of the treatment as very good or good on the GIS, QOLRAD (improvements >60%) and on efficacy, respectively. Poor treatment success was achieved in 13 (GIS), 11 (QOLRAD) and 10 (Efficacy), not necessarily by the same patients. Since about two-thirds of the patients benefited to some extent from the treatment, potato juice in its present formulation may be useful for self treatment. However, placebo-controlled studies with a longer treatment period are needed to confirm this. PMID- 16360928 TI - Traditional Chinese medicines with caspase-inhibitory activity. AB - Clinical evidence indicates that traditional Chinese medicine (TCM) drugs can reduce stroke-inflicted brain damage. To date, the molecular basis of the apparent neuroprotective effects of these TCM drugs remains largely obscure. Several lines of evidence indicate that the activation of cell death programs leads to the loss of neurons during the reperfusion phase of ischemic stroke. In particular, activation of caspases (cysteinyl aspartate-specific proteinases) is a critical step in neuronal apoptosis. Using nuclear magnetic resonance (NMR) and fluorescence assays, we screened a collection of 58 TCM drugs that are commonly used in stroke therapy for caspase inhibitory activity. We found that aqueous extracts of Lianqiao (Fructus Forsythiae) and Shouwuteng (Caulis Polygoni multiflori) blocked the activity of the initiator caspase-8 as well as the effector caspase-3 and caspase-7 in a dose-dependent manner with an IC(50)10 microg/ml. Identification of caspase inhibitory activity of these TCM drugs, allows the formulation of testable hypotheses and design of further investigations aimed at the elucidation of the molecular basis of TCM stroke therapy. PMID- 16360929 TI - Anxiolytic and antidepressant-like activity of a standardized extract from Galphimia glauca. AB - An infusion prepared with aerial parts from Galphimia glauca has been widely used in Mexican traditional medicine as a remedy for nervous excitement. The sedative activity of a methanolic extract from this plant has been demonstrated by neuropharmacological tests. This effect was attributed to the nor-secotriterpene named galphimine B (GB). In the present work, the anxiolytic and antidepressant like effects of G. glauca methanolic extract (standardized on GB content, 8.3mg/g) were assayed by using the elevated plus-maze, light-dark test and the forced swimming paradigm, on ICR albino mice. This extract, administered orally, three times (24, 18 and 1h before the test), and in different doses (125, 250, 500, 1,000 and 2,000 mg/kg) was able to increase significantly (p<0.05) the number of entries, as well as the time spent in the open arms of the elevated plus-maze, indicating an anxiolytic-like effect. A similar effect was observed in the light-dark paradigm test, the time spent in the light box was increased in treated mice. Nevertheless, this treatment was unable to change any parameter in the forced swimming test. Altogether, these results suggest an anxiolytic-like effect to the methanolic standardized extract of G. glauca on ICR inbred mice. PMID- 16360930 TI - Celastrus paniculatus seed oil and organic extracts attenuate hydrogen peroxide- and glutamate-induced injury in embryonic rat forebrain neuronal cells. AB - Seed oil of Celastrus paniculatus Willd. (CP) has been reported to improve memory and the methanolic extract (ME) of CP was shown to exhibit free-radical scavenging properties and anti-oxidant effects in human non-immortalized fibroblasts. In the present study, we have investigated the free-radical scavenging capacity of CP seed oil (CPO) and two extracts, an ethanolic extract (EE) and a ME. CPO and EE showed dose-dependent, free-radical-scavenging capacity, but to a lesser degree than observed for ME. Oxidative stress involves the generation of free radicals and free radical scavenging is one of the mechanisms of neuroprotection. We therefore investigated the effects of CPO, ME, and EE for protection against hydrogen peroxide (H(2)O(2))- and glutamate-induced neurotoxicity in embryonic rat forebrain neuronal cells (FBNC). Pre-treatment of neuronal cells with CPO dose-dependently attenuated H(2)O(2)-induced neuronal death. Pre-treatment with ME and EE partially attenuated H(2)O(2)-induced toxicity, but these extracts were less effective than CPO for neuronal survival. In H(2)O(2)-treated cells, cellular superoxide dismutase (SOD) activity was unaffected, but catalase activity was decreased and levels of malondialdehyde (MDA) were increased. Pre-treatment with CPO, ME, or EE increased catalase activity and decreased MDA levels significantly. Also, CPO pre-treatment attenuated glutamate-induced neuronal death dose-dependently. The activity of cellular acetylcholinesterase (AChE) was not affected by CPO, ME, or EE, suggesting that the neuroprotection offered by CPO was independent of changes in AChE activity. Taken together, the data suggest that CPO, ME, and EE protected neuronal cells against H(2)O(2)-induced toxicity in part by virtue of their antioxidant properties, and their ability to induce antioxidant enzymes. However, CPO, which exhibited the least antioxidant properties, was the most effective in preventing neuronal cells against H(2)O(2)- and glutamate-induced toxicities. Thus, in addition to free-radical scavenging attributes, the mechanism of CP seed component (CP-C) neuroprotection must be elucidated. PMID- 16360931 TI - In vivo anti-osteoporotic activity of isotaxiresinol, a lignan from wood of Taxus yunnanensis. AB - Isotaxiresinol, the main lignan isolated from the water extract of wood of Taxus yunnanensis, was investigated for its effect on bone loss, on serum biochemical markers for bone remodeling and on uterine tissue, using ovariectomized (OVX) rats as the model of postmenopausal osteoporosis. After oral administration of isotaxiresinol (50 and 100mg/kg/d) for 6 weeks, bone mineral content (BMC) and bone mineral density (BMD) in total and cortical bones were increased as compared to those of OVX control rats, and decreases of three bone strength indexes induced by OVX surgery were prevented. Serum biochemical markers for bone remodeling revealed that isotaxiresinol slightly increased bone formation and significantly inhibited bone resorption without side effect on uterine tissue. These results suggest that isotaxiresinol may be useful for treatment of postmenopausal osteoporosis, especially for prevention of bone fracture induced by estrogen deficiency. PMID- 16360932 TI - Activity of Wedelia calendulacea Less. in post-menopausal osteoporosis. AB - Wedelia calendulacea Less., a perennial herb containing isoflavanoids, is used in liver disorders, uterine hemorrhage and menorrhagia. Osteoporosis in women occurs mainly due to estrogen deficiency following menopause. Studies indicate that isoflavones are estrogenic enough to promote bone formation. Our study was aimed to investigate the antiosteoporotic effect of the ethanol extract of W. calendulacea in the ovariectomized rat model of osteoporosis, at two different dose levels of 500 and 750 mg/kg/body wt. day. The findings, assessed on the basis of biomechanical and biochemical parameters, showed that the ethanol extract of the plant had a definite protective effect. This was further supported by the histopathological studies. Phytochemical investigation revealed the presence of isoflavones and wedelolactone, which are known to act as phytoestrogens and may be responsible for the antiosteoporotic activity. PMID- 16360933 TI - Inhibitory effects of aloe carboxypeptidase fraction on streptozotocin-induced enhancement of vascular permeability in the pancreatic islets. AB - The protective actions of components isolated from Aloe arborescens Miller var. natalensis Berger (Kidachi aloe in Japanese) on streptozotocin (Sz)-induced necrosis of B cells in the pancreatic islets of the mouse were investigated to clarify its action mechanism involved in anti-diabetic effects. In this experiment, phenol low molecular weight components of aloin and aloin A that were anti-oxidants and derived from the leaf skin or pulp extract, an aloe carboxypeptidase fraction that is a inhibitor of enhanced vascular permeability and a glycoprotein component that decreases blood glucose were tested with mice precedently administered with Sz which is known as a cytotoxin specific to B cells. The results showed that the treatment group receiving Sz followed by the aloe carboxypeptidase fraction increased the inhibition of dye leakage by 75.8% (p<0.001) in the extract of whole pancreas in comparison to the control group and the aloe carboxypeptidase fraction group also increased the inhibition effect by 68.4% (p<0.001) in the extract of pancreatic islets as compared to the control group. The carboxypeptidase is an aloe-derived protease known to inhibit the acetic acid-related enhancement of intraperitoneal vascular permeability in mice. Further, the elevation of blood glucose in Sz-induced diabetic mice intraperitoneally given the aloe carboxypeptitase fraction was significantly (p<0.01-0.001) restrained at 3, 7 and 14 days after the injection as compared to the control group given solvent only. The results of this experiment suggested that the inhibitory effect on the enhancement of vascular permeability related to the vascular acute inflammatory response at Sz-induced lesions of pancreatic islets was involved in the action mechanism of this enzyme. PMID- 16360934 TI - Bioassay-guided isolation of a vasorelaxant active compound from Kaempferia galanga L. AB - Bioassay-guided fractionation was performed on a crude dichloromethane extract of Kaempferia galanga L. using chromatography techniques. Screening of the extract for biological activity started with the brine shrimp lethality bioassay, followed by the study of its antihypertensive activity on anaesthetized rats, which involved monitoring of the extract's effect on mean arterial blood pressure. The components of the fractions obtained from the separation procedures were analyzed using gas chromatography (GC). The yield of the CH(2)Cl(2) extract was 0.29% of the crude plant extract. Analysis of the data for brine shrimp lethality test using the Finney computer program showed that this extract exhibited potent bioactivity with an ED(50) value of 7.92+/-0.13 microgml(-1). Intravenous administration of the extract induced a dose-related reduction of basal mean arterial pressure (MAP) (130+/-5 mmHg) in the anaesthetized rat, with maximal effects seen after 5-10 min of injection. The gas chromatogram showed that the common compound in the active fractions obtained from the bioassay guided fractionation of the CH(2)Cl(2) extract was ethyl cinnamate. This vasorelaxant active compound, ethyl cinnamate, was isolated as a colorless oil. Ethyl p-methoxycinnamic acid was also isolated as white needles but did not exhibit any relaxant effect on the precontracted thoracic rat aorta. PMID- 16360935 TI - CYP3A4 and CYP2D6 inhibitory activities of Indonesian medicinal plants. AB - Thirty samples of Indonesian medicinal plants were analyzed for their capacity to inhibit in vitro metabolism by human cytochrome P450 3A4 (CYP3A4) and CYP2D6 with a radiometric assay. The MeOH-soluble fractions of 25 samples, prepared from water extracts, demonstrated inhibitory activity more than 50% on the metabolism mediated by CYP3A4, and 21 samples on the metabolism mediated by CYP2D6. Among the MeOH-soluble fractions, Piper nigrum leaf showed the highest inhibitory activity against CYP3A4 (91.7%), and Punica granatum against CYP2D6 (98.1%). The water extracts of which MeOH-soluble fraction showed inhibitory activity more than 70% were fractionated with EtOAc. From the EtOAc-soluble fractions, Curcuma heyneana (67.0%), Pi. cubeba (75.0%), Pi. nigrum fruit (84.0%), Pi. nigrum leaf (85.8%), and Zingiber aromaticum (75.3%) demonstrated inhibitory activity more than 50% on the metabolism mediated by CYP3A4, but only Pi. nigrum fruit (72.8%) and Pi. nigrum leaf (69.1%) showed strong inhibitory activity against CYP2D6. For samples that showed more than 70% inhibition, their IC(50) values were determined. The most potent inhibitory activity against CYP3A4 (IC(50) value of 25 microg/ml) was found for the extract of Pi. nigrum leaf, while that of Catharanthus roseus showed the most potent inhibitory effect against CYP2D6 (IC(50) value of 11 microg/ml). These results should indicate once more the possibility of potential medicinal plant-drug interactions. PMID- 16360936 TI - Tinospora cordifolia induces enzymes of carcinogen/drug metabolism and antioxidant system, and inhibits lipid peroxidation in mice. AB - The present study is an effort to identify a potent chemopreventive agent against various diseases (including cancer) in which oxidative stress plays an important causative role. Here, we investigated the effect of a hydroalcoholic (80% ethanol: 20% distilled water) extract of aerial roots of Tinospora cordifolia (50 and 100mg/kg body wt./day for 2 weeks) on carcinogen/drug metabolizing phase-I and phase-II enzymes, antioxidant enzymes, glutathione (GSH) content, lactate dehydrogenase and lipid peroxidation in liver of 8-week-old Swiss albino mice. The modulatory effect of the extract was also examined on extrahepatic organs, i.e., lung, kidney and forestomach, for the activities of GSH S-transferase (GST), DT-diaphorase (DTD), superoxide dismutase (SOD) and catalase. Significant increases in the levels of acid-soluble sulfhydryl (-SH) and cytochrome P(450) contents, and enzyme activities of cytochrome P(450) reductase, cytochrome b(5) reductase, GST, DTD, SOD, catalase, GSH peroxidase (GPX) and GSH reductase (GR) were observed in the liver. Both treated groups showed decreased malondialdehyde (MDA) formation. In lung SOD, catalase and GST; in kidney SOD and catalase; and in forestomach SOD, DTD and GST showed significant increase at both dose levels of treatment. BHA (0.75%, w/w in diet), a pure antioxidant compound, was used as a positive control. This group showed increase in hepatic levels of GSH content, cytochrome b(5), DTD, GST, GR and catalase, whereas MDA formation was inhibited significantly. In the BHA-treated group, the lung and kidney showed increased levels of catalase, DTD and GST, whereas SOD was significantly increased in the kidney and forestomach; the latter also showed an increase in the activities of DTD and GST. The enhanced GSH level and enzyme activities involved in xenobiotic metabolism and maintaining antioxidant status of cells are suggestive of a chemopreventive efficacy of T. cordifolia against chemotoxicity, including carcinogenicity, which warrants further investigation of active principle (s) present in the extract responsible for the observed effects employing various carcinogenesis models. PMID- 16360937 TI - Structure prerequisite for antioxidant activity of silybin in different biochemical systems in vitro. AB - Structural analogues (flavanone: 2-4 and flavone: 5 and 6, respectively) of silybin (1a) were synthesized and tested for inhibitory activity on O(2)(-) release and PKC translocation in PMA-stimulated neutrophils as well as xanthine oxidase activity in order to identify the molecular structures responsible for the antioxidant property of silybin. Concerning the prevention of hem-mediated oxidative modification of LDL by silybin, the hydroxyl radical scavenging activity of its structural analogues was also determined. We demonstrated that the basic skeleton of 1a (4) is responsible for its inhibitory activity on O(2)( ) release in PMA-stimulated neutrophils via inhibition of PKC translocation, since introduction of a double bound and hydroxyl groups at C-5 and C-7 position (5 and 6) did not result in further increase in inhibition of O(2)(-) release. It has been shown that the presence of the phenolic hydroxyl group at C-5 and C-7 of 1a is essential for the inhibition of xanthine oxidase activity. Moreover, introduction of a double bond into the C-ring of 2 and 3, resulting in flavone derivatives (5 and 6), markedly enhanced the antioxidant effect in all the tested systems. Finally, silybin (1a) and its flavon derivatives (5 and 6) directly scavenged hydroxyl radicals as well. On the basis of these results it might be concluded that different moiety of silybin is responsible for inhibition of overproduction of O(2)(-) in stimulated neutrophils, xanthine oxidase activity, and for prevention of hem-mediated oxidative modification of LDL. PMID- 16360938 TI - Effect of "Dang-Qui-Shao-Yao-San" a Chinese medicinal prescription for dysmenorrhea on uterus contractility in vitro. AB - Dysmenorrhea is directly related to elevated level of prostaglandin F(2)alpha (PGF(2)alpha), and is treated with nonsteroid anti-inflammatory drugs. Though efficacy of the latter is rapid, the effect is temporary, and they cause many side effects to the liver, kidney and digestive system. Because of this, Chinese medicinal therapy is being considered as a feasible alternative medicine. In this study, Dang-Qui-Shao-Yao-San (DQSYS), one of Chinese medicinal prescriptions was selected. A 50% alcohol solution was used to extract the active ingredients and create a freeze-dried product. Using rat uterine smooth muscle in vitro, DQSYS was used to suppress spontaneous contractions and PGF(2)alpha induced contractions. Then acetylcholine, ergonovine, propranolol, oxytocin and KCl were used to analyze the physiological mechanisms. The results show that DQSYS has the antagonistic action on both PGF(2)alpha and Ach induced uterine contraction. Furthermore, it has the antagonistic effect on contraction caused by KCl depolarization. These actions may be the major mechanism of the beneficial effect of Chinese prescription in treating dysmenorrhea. PMID- 16360939 TI - Protective effects of Rosmarinus tomentosus ethanol extract on thioacetamide induced liver cirrhosis in rats. AB - The capability of an ethanol extract of Rosmarinus tomentosus to protect rat liver in an experimental model of cirrhosis induced by thioacetamide (TAA) has been evaluated. Four groups of rats were used: Two of them received 300 mg TAA/l in the drinking water for 3 months while the other two, which served as controls, were given water ad libitum. During the same period and for each one of the treatments, one group received a semi-purified (SP) diet and the other one was fed the same diet supplemented with 1% of the dry residue obtained from R. tomentosus ethanol extract (SP+E). There was a significant reduction of TAA toxicity in rats fed the SP+E diet, as assessed by plasma and liver biochemical markers, and by liver histopathology. Plasma total protein concentration was restored, urea concentration and plasma alkaline phosphatase and gamma-glutamyl transferase activities were reduced. A significant correction of plasma fatty acids concentrations was also evident. Hepatic alkaline phosphatase and gamma glutamyl-transferase activities were significantly reduced in animals fed SP+E diet and glucose-6-phosphatase activity was significantly enhanced. The results suggest that R. tomentosus ethanol extract administered in the diet affords protection against TAA-induced cirrhosis, preventing most of the histological changes and functionality alterations own to this experimental pathology. PMID- 16360940 TI - Hypoglycemic effects of the wood of Taxus yunnanensis on streptozotocin-induced diabetic rats and its active components. AB - Hypoglycemic effects of the H(2)O and MeOH extracts of the wood of Taxus yunnanensis were examined in streptozotocin (STZ)-induced diabetic rats. The H(2)O extract significantly lowered the fasting blood glucose level by 33.7% at a 100mg/kg dose on intraperitoneal administration. From the active H(2)O extract of the wood, three lignans, i.e., isotaxiresinol (1), secoisolariciresinol (2) and taxiresinol (3), were isolated as major components. These lignans were further tested for their hypoglycemic effects on the same experimental model. At a dose of 100mg/kg (i.p.), isotaxiresinol (1) reduced the fasting blood glucose level of diabetic rats by 34.5%, while secoisolariciresinol (2) and taxiresinol (3) reduced by 33.4% and 20.9%, respectively. The blood glucose lowering effects of 1 and 2 were stronger than the mixture of tolbutamide (200mg/kg) and buformin (1mg/kg) used as a positive control, which lowered fasting blood glucose level by 24.0%. PMID- 16360941 TI - Muscle damage induced by black cohosh (Cimicifuga racemosa). AB - Extracts of black cohosh (Cimicifuga racemosa) are commonly used for the treatment of symptoms associated with menopause. Adverse events with black cohosh are rare, mild and reversible. A few number of serious adverse events, including hepatic and circulatory conditions, have been also reported, but without a clear causality relationship. We report the case of a woman with severe asthenia and very high blood levels of creatine phosphokinase and lactate dehydrogenase. The patient referred to take a dietary supplement derived from black cohosh for ameliorating menopause vasomotor symptoms. To exclude a possible involvement of this product, the patient was suggested to discontinue this therapy. After suspicion the patient showed a progressive normalization of biochemical parameters and improvement of clinical symptoms. We can hypothesise a causative role for black cohosh in the muscle damage observed in this patient. Factors suggesting an association between black cohosh and the observed myopathy included the temporal relationship between use of herbal product and asthenia and the absence of other identified causative factors. Rechallenge with the suspected agent was inadvisable for ethic reasons because of the risk of a serious relapse. This is the first time that asthenia associated with high muscle enzymes serum levels by black cohosh has been reported. In our opinion, this report is of interest because of the widespread diffusion of use of black cohosh as an alternative medicine for relief from menopausal symptoms. PMID- 16360943 TI - Repelling properties of some plant materials on the tick Ixodes ricinus L. AB - The repellent effects on nymphal stages of Ixodes ricinus L. of some plant materials have been studied in the laboratory. The plant material consisted of an ethanolic extract from Achillea millefolium L., and volatile oils of birch and/or pine tar, citronella, cloves, eucalyptus, geranium, lavender, lily of the valley and peppermint. The most pronounced effects were observed for the oils of citronella, cloves and lily of the valley. They possessed repelling activities of the same magnitude as the reference repellent DEET (N,N-diethyl-m-toluamide). Some major constituents of these oils, e.g. citronellol and geraniol (oil of citronella and lily of the valley) and eugenol (oil of cloves) showed pronounced repelling effects. This was also the case for phenethyl alcohol, a minor component in the oil from lily of the valley. PMID- 16360942 TI - The pharmacognosy of Humulus lupulus L. (hops) with an emphasis on estrogenic properties. AB - As the population ages, there is an ever-increasing need for therapeutic agents that can be used safely and efficaciously to manage symptoms related to postmenopausal estrogen deficiency. Endogenous estrogens, e.g., 17beta-estradiol, of exogenous mammalian origin, e.g., horses, have long been used to manage such symptoms. There are more than 20 different classes of phytochemicals that have demonstrated affinity for human estrogen receptors in vitro. Some studies on exogenous estrogenic substances of botanical origin (phytoestrogens), such as standardized formulations of plant extracts with in vitro and in vivo estrogenic activity from soy (Glycine max Merill.) and red clover (Trifolium pratense L.), suggest clinical efficacy. Few clinical data for phytoestrogens other than isoflavonoids are available. In an exhaustive review of the literature through 2003, only two clinical trials were identified that were designed to evaluate the effect of hops (Humulus lupulus L.) on symptoms related to menopause. Folkloric, chemical, and biological literature relating primarily to the use of hops for their estrogenic activity, and two human clinical trials, are reviewed. PMID- 16360944 TI - Regulatory mechanisms of chitin biosynthesis and roles of chitin in peritrophic matrix formation in the midgut of adult Aedes aegypti. AB - In mosquitoes, the peritrophic matrix is formed in response to blood feeding and can be a physical barrier when pathogens ingested with blood meal attempt to reach and transverse the midgut epithelium. The main components of the peritrophic matrix are chitin-biding-domain containing proteins, glycosylated proteins, and chitin fibrils. Chitin is synthesized from fructose-6-phosphate by a series of five enzymatic reactions. We previously found that blood feeding induces transcriptional up-regulation of glutamine: fructose-6-phosphate amidotransferase-1 (AeGfat-1) and chitin synthase (AeCs), the first and last enzymes of the biosynthetic pathway, respectively, in the midgut of Aedes aegypti. In this study, we demonstrated that formation of the peritrophic matrix is disrupted when the transcript abundance of either gene is knocked-down using RNAi methodologies. We also have shown that enzymatic activity of recombinant AeGFAT-1 is sensitive to feedback inhibition by UDP-N-acetylglucosamine, a substrate of chitin synthase. These findings demonstrate that in the midgut of adult Ae. aegypti, (1) chitin is synthesized de novo in response to blood feeding and is an essential component of the peritrophic matrix, and (2) chitin biosynthesis is negatively regulated, in part, by inhibitory sensitivity of AeGFAT-1 to UDP-N-acetylglucosamine. PMID- 16360945 TI - Phylogenetic shadowing of a histamine-gated chloride channel involved in insect vision. AB - A recently identified gene, hclA (synonym: ort), codes for an ionotrophic histamine receptor subunit in Drosophila melanogaster, and known hclA mutations lead to defects in the visual system, neurologic disorders and changed responsiveness to neurotoxins. To investigate whether this novel class of receptors is common across the Insecta, we analysed the genomes of 15 other insect species (Diptera, Hymenoptera, Coleoptera, Lepidoptera) and revealed orthologs of hclA in all of them. The predicted receptor domain of HCLA is extensively conserved (86-100% of identity) among the 16 proteins. Minor changes in the amino acid sequence that includes the putative transmembrane domains (TMs) 1-3 were found in non-drosophilid species only. Substantial amino acid variability was observed in the signal polypeptides, the intracellular loop domains and in TM4, in good accordance with known data on sequence variations in ligand-gated ion channels. Pairwise comparisons revealed three consensus sequences for N-glycosylation, conserved in HCLAs of all species studied, as well as a drosophilid-specific putative phosphorylation site. Real-time PCR analysis demonstrated that hclA-mRNA is abundant in heads of adult Drosophila. However, species- and sex-specific variations of the hclA expression levels were also observed. PMID- 16360946 TI - Insect freeze tolerance: Roles of protein phosphatases and protein kinase A. AB - Freeze-tolerant larvae of the goldenrod gall fly, Eurosta solidaginis Fitch, show multiple metabolic adaptations for subzero survival including the autumn synthesis of high concentrations of polyols. The induction and regulation of cold hardiness adaptations requires the intermediary action of signal transduction enzymes. The present study evaluates changes in the activities of cAMP-dependent protein kinase (PKA), protein phosphatases 1 (PP1), 2A, 2C, and protein tyrosine phosphatases (PTPs) over the course of the winter season and also in insects exposed to -4, -20 degrees C, or anoxic conditions in the laboratory. The increased PKA and decreased PP1 over the winter season and/or at subzero temperature support a regulatory role for these enzymes in cryoprotectant polyol synthesis. PTP activities were also strongly increased under these conditions and may act to antagonize tyrosine kinase mediated cell growth and proliferation responses and, thereby, contribute to hypometabolism and diapause over the winter. PMID- 16360947 TI - Molecular cloning and functional characterization of an aspartic protease from the hard tick Haemaphysalis longicornis. AB - Haemaphysalis longicornis cDNA encoding an aspartic protease (longepsin) was identified from a midgut cDNA library. The longepsin cDNA contains 1176bp that code for 392 amino acid residues with a predictable molecular weight of 39.3kDa. The cDNA has a signal peptide sequence associated with the N-terminal domains and domain structure analysis revealed that the deduced protein has two aspartic acid residues that are characteristic of a single active site for aspartic proteases. This novel longepsin cDNA exhibits 57% identity to the lysosomal aspartic protease of Aedes aegypti, 52% to Bombyx mori cathepsin D, 38% to Ancylostoma caninum, 44% to Schistosoma mansoni and 28% to Boophilus microplus aspartic proteases. The DNA fragment coding for longepsin was cloned into a pGEX-4T-3 vector and expressed in Escherichia coli. The recombinant longepsin, once activated was able to hydrolyze casein substrate as well as hemoglobin (Hb) under acidic conditions (pH 3.5). RT-PCR analysis showed that the longepsin mRNA transcripts were expressed in salivary glands and midgut and not in the ovary. Northern blot analysis revealed that longepsin (1.5kb) was expressed in unfed and partially fed ticks and expression levels increased during feeding. The finding that longepsin is expressed in the midgut and salivary glands, proteolytic activity occurs under acidic conditions and longepsin can be gene silenced of longepsin provides compelling support for the hypothesis that longepsin plays an integral role in the proteolysis of erythrocyte Hb obtained from a host blood meal. PMID- 16360948 TI - Immune challenge induces N-terminal cleavage of the Drosophila serpin Necrotic. AB - The Drosophila Necrotic protein is a serine proteinase inhibitor, which regulates the Toll-mediated innate immune response. Necrotic specifically inhibits an extracellular serine proteinase cascade leading to activation of the Toll ligand, Spatzle. Necrotic carries a polyglutamine extension amino-terminal to the core serpin structure. We show here that cleavage of this N-terminal extension occurs following immune challenge. This modification is blocked in PGRP-SA(semmelweiss) mutants after Gram-positive bacterial challenge and in persephone mutants after fungal or Gram-positive bacterial challenge, indicating that activation of either of the Toll pathway upstream branches induces N-terminal cleavage of the serpin. The absolute requirement of persephone gene product for this cleavage indicates that Gram-positive bacteria activate a redundant set of proteinases upstream of Toll. Both full-length Necrotic and the core serpin are active inhibitors of a range of serine proteinases: the highest affinity being for cathepsin G and elastases. We found a 13-fold increase in the specificity of the core serpin over that of full-length Necrotic for one of the tested proteinases (porcine pancreatic elastase). This finding indicates that cleavage of the Necrotic amino terminal extension might modulate Toll activation following the initial immune response. PMID- 16360949 TI - The roles of ERK and P38 MAPK signaling cascades on embryonic diapause initiation and termination of the silkworm, Bombyx mori. AB - To clarify the molecular mechanisms of Bombyx diapause, we focused on mitogen activated protein kinases (MAPK), which are the major components of signal transduction cascades that regulate cell proliferation, differentiation, and stress responses. In the present study, cloning of Bombyx extracellular signal related kinase (ERK), MAPK-ERK kinase (MEK), and p38 MAPK cDNA, revealed that their amino acid sequences have close similarity with those of other species. We analyzed the roles of the kinases in diapause initiation and termination by immuno-blotting with anti-phospho-kinase antibodies. Phospho-MEK levels remained consistently high in non-diapausing eggs, then declined after the diapausing stage in diapausing eggs, and began to increase 45 d after transfer to 5 degrees C upon diapause termination. The phospho-ERK and phospho-MEK profiles were similar, suggesting that ERK phosphorylation is regulated by MEK. The phospho-p38 MAPK levels declined 36 h after oviposition in diapausing eggs, and increased at 15-30 d at 5 degrees C in yolk cells, suggesting that p38 MAPK has a role in diapause initiation and termination. Phospho-ERK levels were maintained with diapause-interrupting treatment and declined with diapause-sustaining treatment. ERK phosphorylation is considered to have a role in diapause termination and in the resumption of development. PMID- 16360950 TI - cDNA sequences of two biliproteins, BP1 and BP2, from the cabbage white butterfly, Pieris rapae and their tissue- and stage-specific accumulation. AB - Two similar full-length cDNAs of biliprotein were isolated and shown to encode the two isoelectric forms of biliverdin-binding proteins (BPs): BP1 and BP2 in Pieris rapae. Sequence analysis of two cDNA clones shows that both BPs contain a 567-bp open reading frame which predicts a 189-amino acid protein and a 15-amino acid signal peptide. The calculated isoelectric points are pI=7.25 (BP1) and 6.74 (BP2), respectively. Comparison of two sequences of BP1 and BP2 reveals 12 base differences in the open reading frame, of which three nucleotide changes lead to two amino acid substitutions. The 5'-UTR from the two clones shows no difference, but an additional 45-bp fragment is inserted in the 3'-UTR of BP2 making its message a little longer than that of BP1. Northern blot analysis confirmed that the BP mRNAs are expressed from the late 4th instar to the adult stage with exception of prepupae and newly ecdysed pupae. While the BP1 transcript was prevalent in the larval stage, the BP2 transcript was abundant in the whole body only after the pupal stage in P. rapae. Both BPs were detected in a stage specific pattern in the epidermis, testis, hindgut, wing, brain, and egg, with a lesser amount in the fat body. Two-dimensional gel electrophoresis and Western blot analyses revealed that BP1 was dominant in tissues from larvae, BP2 was dominant in tissues from pupal stages, and both BPs appeared in tissues from the adult stage, though BP2 was predominant. PMID- 16360951 TI - Expression of one isoform of GTP cyclohydrolase I coincides with the larval black markings of the swallowtail butterfly, Papilio xuthus. AB - The larva of the swallowtail butterfly Papilio xuthus changes its body markings during the fourth ecdysis. We found that stage-specific cuticular black markings are mainly regulated by co-localization of two melanin synthesis enzymes; tyrosine hydroxylase (TH) and dopa decarboxylase (DDC). TH converts tyrosine to dihydroxyphenylalanine (dopa), and tyrosine itself is converted from phenylalanine by phenylalanine hydroxylase (PAH). Guanosine triphosphate cyclohydrolase I (GTPCHI) is essential for the synthesis of tetrahydrobiopterin (BH4) that is a cofactor of TH and PAH. In this report, we found that a GTPCHI inhibitor prevents pigmentation in cultured integuments, suggesting that the GTPCHI activity is also involved in cuticle pigmentation. We have cloned GTPCHI and PAH cDNAs from P. xuthus and investigated their spatial expression patterns in epidermis by whole-mount in situ hybridization. There are two isoforms of GTPCHI in larval epidermis (GTPCHIa and GTPCHIb). GTPCHIa is expressed at the black markings of the subsequent instar, similar to TH, whereas GTPCHIb is expressed uniformly, similar to PAH. This suggests that the region-specific expression of GTPCHIa supplies sufficient BH(4) reinforcing the TH activity in black marking area. Our results imply that larval markings are regulated by not only melanin synthesis enzymes but also the cofactor supplying enzyme. PMID- 16360952 TI - Multiple origins of pyrethroid resistance in sympatric biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae). AB - The two most damaging biotypes of Bemisia tabaci, B and Q, are sympatric in the Mediterranean basin and show high resistance to pyrethroids synergized by organophosphates. Previous work showed that in the B biotype, this resistance is associated with the L925I mutation in the para-type voltage gated sodium channel. Here we identified two mutations in the para-type voltage gated sodium channel associated with resistance to pyrethroids synergized by organophosphates in the Q biotype: the L925I mutation that occurs in the B biotype, and substitution of threonine to valine, at position 929 (T929V). To determine if the L925I and T929V mutations have single or multiple origins, we sequenced the DNA regions flanking the mutations from 13 B and Q strains collected worldwide. The survey identified five resistant alleles and five susceptible alleles. In the resistant alleles, the nucleotide diversity was low within biotypes (0.001), but high between biotypes (0.033). Nucleotide diversity in susceptible alleles was high between the two biotypes (0.028). These observations are consistent with multiple independent origins of resistance. Although the B and Q biotypes coexist in several regions of the Mediterranean basin, divergence in their DNA sequences at the para-type voltage gated sodium channel locus suggests gene flow between these biotypes is low or nil. PMID- 16360953 TI - WO bacteriophage transcription in Wolbachia-infected Culex pipiens. AB - Bacteriophages are commonly found in association with free-living bacteria, both as exogenic phages (virions) and as prophages integrated into the bacterial genome. In contrast, the observation of bacteriophages associated with obligate intracellular bacteria has been described infrequently. An exception is provided by Wolbachia endosymbionts, which harbor multiple phage elements that have been designated as WO phage. Wolbachia are maternally inherited bacteria that occur in the cytoplasm of many invertebrates, where they often manipulate host reproduction. Previously, the WO phage orf7 locus and ankyrin repeat-encoding genes have been observed to represent sources of genetic diversity between Wolbachia (wPip) strains infecting mosquitoes of the Culex pipiens complex and have been suggested as potential participants in the reproductive manipulations. We have characterized WO phage associated with multiple Wolbachia-infected Culex strains and an uninfected strain using electron microscopy and RT-PCR. For each strain, different developmental stages were examined for transcription of three WO phage orf7 genes. The results provide evidence for the presence of both actively transcribed virions and inactive prophages. Variable orf7 transcription patterns are observed in comparisons of differing Cx. pipiens strains. Variability includes both mosquito stage-specific and sexually dimorphic orf7 expression patterns. This report provides additional support for the hypothesis that bacteriophages play an important role in Wolbachia and host evolution. PMID- 16360954 TI - Molecular characterisation of nicotinic acetylcholine receptor subunits from the cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae). AB - As part of a program to monitor the susceptibility of cat flea populations to the insecticide imidacloprid we have examined the cat flea nicotinic acetylcholine receptor, the target site protein of the neonicotinoid group of insecticides. Seven nAChR subunits (six alpha-type and one beta-type) were identified in cat flea using a degenerate PCR-based strategy. Five of these were expressed in vitro by creating chimeras containing the N-terminal ligand-binding domain of the cat flea subunits and the C-terminal region of the Drosophila Dalpha2 (SAD) subunit. Two of the five chimeric subunits, Cfalpha1/Dalpha2 and Cfalpha3/Dalpha2, when co expressed with rat beta2 in Drosophila S2 cells, showed high-affinity binding of both epibatidine (Kd=1.6+/-0.6 and 0.13+/-0.06nM, respectively), and imidacloprid (Ki=142+/-34 and 28.7+/-2.4nM, respectively). It is likely therefore that Cfalpha1 and Cfalpha3 contribute to nAChR populations in vivo that are sensitive to imidacloprid. The identification of cat flea nAChR subunits that have a high affinity for imidacloprid presents candidate genes in which to look for resistance-associated mutations if target-site resistance to imidacloprid arises in domestic pet flea populations. PMID- 16360955 TI - Electrochemically controlled release of dexamethasone from conducting polymer polypyrrole coated electrode. AB - Chronic recordings from micromachined neural electrode arrays often fail a few weeks after implantation primarily due to the formation of an astro-glial sheath around the implant. We propose a drug delivery system, from conducting polymer (CP) coatings on the electrode sites, to modulate the inflammatory implant-host tissue reaction. In this study, polypyrrole (PPy) based coatings for electrically controlled and local delivery of the ionic form of an anti-inflammatory drug, dexamethasone (Dex), was investigated. The drug was incorporated in PPy via electropolymerization of pyrrole and released in PBS using cyclic voltammetry (CV). FTIR analysis of the surface showed the presence of Dex and polypyrrole on the coated electrode. The thickness of the coated film was estimated to be approximately 50 nm by ellipsometry. We are able to release 0.5 mug/cm(2) Dex in 1 CV cycle and a total of almost 16 mug/cm(2) Dex after 30 CV cycles. In vitro studies and immunocytochemistry on murine glial cells suggest that the released drug lowers the count of reactive astrocytes to the same extent as the added drug. In addition, the released drug is not toxic to neurons as seen by healthy neuronal viability in the released drug treated cells. PMID- 16360956 TI - Encapsulation of the immune potentiators MPL and RC529 in PLG microparticles enhances their potency. AB - PURPOSE: Monophosphoryl lipid A (MPL) and the synthetic LPS mimetic RC529, encapsulated in poly(lactide-co-glycolide) (PLG) microparticles, were evaluated as immune potentiators in the presence of either HIV-1 gp120 protein or antigen from Neisseria meningitidis serotype B (Men B). The immunogenicity of these formulations was evaluated in mice and compared to CpG containing oligonucleotide. This work was done as part of an ongoing effort to enhance the potency of vaccine candidates against HIV and Men B. METHODS: Microparticles were made by a solvent evaporation method. Blank microparticles as well as microparticles with encapsulated MPL or RC529 were made using the PLG polymer RG503 and the ionic surfactant Dioctylsulfosuccinate by the water-in-oil-in-water emulsion technique. Antigens from HIV-1 and Men B were adsorbed on the surface of these anionic microparticles and the final formulations characterized for protein loading, release, and integrity. The formulations were then tested in mice for their ability to elicit antibodies and bactericidal activity in comparison with CpG containing oligonucleotide. RESULTS: We have found that adding soluble immune potentiators to Men B antigen formulated on PLG microparticles significantly enhanced the immune response to a level comparable to that obtained using CpG. In a separate study, we found that encapsulating MPL or RC529 in PLG microparticles further enhanced the response in comparison to soluble CpG, which is our control group. Similarly, adding soluble immune potentiators to gp120 antigen formulated on PLG microparticles resulted in a significant enhancement of the immune response. Moreover, delivering MPL or RC529 encapsulated in PLG microparticles with gp120 adsorbed on PLG microparticles, resulted in even further enhancement of serum titers over those obtained with soluble immune potentiators. These titers were comparable to or greater than those obtained with soluble CpG, the control group. This effect was observed for both antigens regardless of whether or not the immune potentiator and the antigen were used with the same or with separate particles. In conclusion, the advantages of encapsulating MPL and RC529 lie not only in the enhanced immune response they elicit, but also in the convenience of handling these relatively insoluble compounds, and flexibility in vaccine design. The fact that MPL and RC529 are readily soluble in methylene chloride used for the manufacturing of PLG microparticles makes it easy to avoid solubility issues. Moreover, formulating antigen and immune potentiator with the same particle offers an attractive approach to vaccine delivery. PMID- 16360957 TI - Induction of apoptosis in A549 human lung cancer cells by all-trans retinoic acid incorporated in DOTAP/cholesterol liposomes. AB - All-trans retinoic acid (ATRA) has been shown to exert anti-cancer activities in a number of types of cancer cells. However, it has been reported that many NSCLC exhibited resistance to ATRA treatment. In the present study, we hypothesized that intracellular delivery of ATRA would overcome the ATRA resistance in A549 cells. Here, we investigated the induction of apoptosis by ATRA incorporated in cationic liposomes composed of DOTAP/cholesterol in A549 human lung cancer cells, which are insensitive (resistant) to the growth inhibitory effects of ATRA. The zeta potentials of DOTAP/cholesterol liposomes and DSPC/cholesterol liposomes were about +50 and -3 mV. In A549 cells, [(3)H]ATRA incorporated in DOTAP liposomes showed increased cellular association compared with [(3)H]ATRA or [(3)H]ATRA incorporated in DSPC/cholesterol liposomes. ATRA incorporated in DOTAP/cholesterol liposomes showed much higher cytotoxic effects and apoptosis inducing activity compared with ATRA or ATRA incorporated in DSPC/cholesterol liposomes. The enhanced expression of TIG3 mRNA tumor suppressor gene by ATRA incorporation into DOTAP/cholesterol liposomes might partly explain the mechanism of enhanced cytotoxicity and/or apoptosis. These observations provide valuable information to help in the design of differentiation therapy by ATRA in non-small cell lung carcinoma. PMID- 16360958 TI - Economic implications of extended-release metoprolol succinate for heart failure in the MERIT-HF trial: a US perspective of the MERIT-HF trial. AB - BACKGROUND: The MERIT-HF trial demonstrated improved survival and fewer hospitalizations for worsening heart failure with extended-release (ER) metoprolol succinate in patients with heart failure. This study sought to estimate the economic implications of this trial from a US perspective. METHODS AND RESULTS: A discrete event simulation was developed to examine the course of patients with heart failure. Characteristics of the population modeled, probabilities of hospitalization and death with standard therapy, and risk reductions with ER metoprolol succinate were obtained from Metoprolol CR/XL Randomized Intervention Trial in Chronic Heart Failure (MERIT-HF) and evaluated in weekly cycles. Direct medical costs were estimated from US databases in 2001 US dollars. Uncertainty in inputs was incorporated and analyses were carried out to estimate events prevented total and net costs. The model predicts that ER metoprolol succinate will prevent approximately 7 deaths and 15 hospitalizations from heart failure per 100 patients over 2 years. Compared with standard therapy alone, this translates to a cost reduction between $395 and $1112 per patient, depending on whether the costs of hospitalizations for other causes are included. Savings were maintained in 90% of the simulations. CONCLUSION: This analysis predicts that the positive effect of ER metoprolol succinate on mortality and morbidity demonstrated in MERIT-HF leads to substantial savings. PMID- 16360959 TI - Will beta-blockers save money? PMID- 16360960 TI - Rationale and design of the enoximone clinical trials program. AB - BACKGROUND: Chronic heart failure is a disease syndrome characterized in its advanced stages by a poor quality of life, frequent hospitalizations, and a high risk of mortality. In advanced and ultra-advanced chronic heart failure, many treatment options, such as cardiac transplantation and mechanical devices, are severely limited by availability and cost. Short-term Phase II clinical trials suggest that low-dose oral inotropic therapy with enoximone may improve hemodynamics and exercise capacity, without adversely affecting mortality, in selected subjects with advanced chronic heart failure. Based on these data, the ability of enoximone to deliver safe and efficacious palliative treatment of advanced/ultra-advanced chronic heart failure is being evaluated in Phase III clinical trials. METHODS AND RESULTS: The Enoximone Clinical Trials Program is a series of 4 clinical trials designed to evaluate the safety and efficacy of oral enoximone in advanced chronic heart failure. ESSENTIAL I and II (The Studies of Oral Enoximone Therapy in Advanced Heart Failure) will investigate the effects of oral enoximone on all-cause mortality and cardiovascular hospitalization, submaximal exercise capacity, and quality of life in subjects with New York Heart Association Class III/IV chronic heart failure. EMOTE (Oral Enoximone in Intravenous Inotrope-Dependent Subjects) will evaluate the potential of oral enoximone to wean subjects with ultra-advanced chronic heart failure from chronic intravenous inotropic therapy to which they have been shown to be dependent. EMPOWER (Enoximone Plus Extended-Release Metoprolol Succinate in Subjects with Advanced Chronic Heart Failure) will explore the potential of enoximone to increase the tolerability of continuous release metoprolol in subjects shown previously to be hemodynamically intolerant to beta-blocker treatment. CONCLUSION: These studies are Phase III, multicenter, randomized, double-blinded, placebo-controlled trials designed to test the general hypothesis that chronic oral administration of low doses of enoximone can produce beneficial effects in subjects with advanced or ultra-advanced chronic heart failure. PMID- 16360961 TI - Surgical alternatives for the palliation of heart failure: a prospectus. AB - BACKGROUND: Congestive heart failure (CHF) is the leading cause of hospital admissions in the United States. METHODS AND RESULTS: CHF has a variety of palliative options for treatment and 1 curative one: cardiac transplantation. Palliative medical therapies are often limited in effectiveness by progression of the disease or patient intolerance. Because of limited donor availability, alternative surgical strategies are now being relied on for palliation of patients in end-stage CHF. CONCLUSION: In this manuscript, we review the principles, outcomes, and practices of some of these surgical strategies often used in the palliation of end-stage CHF. PMID- 16360962 TI - The role of a common adenosine monophosphate deaminase (AMPD)-1 polymorphism in outcomes of ischemic and nonischemic heart failure. AB - BACKGROUND: A common variant of the adenosine monophosphate deaminase (AMPD)-1 gene (C34T) results in enzymatic inactivity and may increase adenosine in cardiac muscle and confer cardioprotection through ischemic preconditioning. METHODS AND RESULTS: We hypothesized that AMPD1 carriers with ischemic heart failure (HF) in the Beta-Blocker Evaluation of Survival Trial (BEST) might have a relative survival advantage. Patients (n = 1038, 20% black) with ischemic (58%) and nonischemic (42%) HF were followed for an average of 2.0 years for cardiovascular mortality. DNA was purified from blood using phenol/chloroform. Genotyping was performed by polymerase chain reaction with 5' exonuclease chemistry. Differences in survival were assessed by comparing Kaplan-Meier curves with the log-rank test. Genotype frequencies differed by ethnicity (P < .001) but not by disease etiology. AMPD1 genotype did not significantly modify survival in the entire study population (hazard ratio [HR] = 0.91, 95% CI = 0.61-1.37), among ischemics (HR = 0.73, CI = 0.44-1.22, P = .23), or ischemic non-blacks (HR = 0.74, CI = 0.44-1.24, P = .25). Genotype did not modify the effect of bucindolol on mortality. CONCLUSIONS: Results of this study failed to confirm a reported survival benefit among HF patients carrying the AMPD1 T-allele. However, further studies in larger, more homogeneous populations should explore the possibility of a modest survival advantage for patients with ischemic HF. PMID- 16360964 TI - Does "anti-lipid" also mean "anti-arrhythmic"? PMID- 16360963 TI - Atorvastatin therapy increases heart rate variability, decreases QT variability, and shortens QTc interval duration in patients with advanced chronic heart failure. AB - BACKGROUND: Although statins decrease the incidence of ventricular arrhythmias in patients with atherosclerotic heart disease, their potential antiarrhythmic effects in heart failure remain undefined. METHODS AND RESULTS: Of 80 heart failure patients enrolled, 40 were randomized to receive atorvastatin (statin group); the remaining 40 served as controls. At baseline and after 3 months, we measured heart rate variability (HRV), QT variability (QTV), and QTc interval using interactive high-resolution electrocardiogram analysis. The 2 groups did not differ in baseline HRV standard deviation of normal-to-normal intervals (SDNN) (RR): 24.6 +/- 2.8 ms in statin group versus 24.8 +/- 3.1 ms in controls, P = .72; square root of the mean of squared differences between successive intervals (rMSSD) (RR): 21.2 +/- 2.7 ms versus 21.7 +/- 2.9 ms, P = .43), QTV SDNN (QT): 6.4 +/- 1.5 ms versus 6.4+/-1.7, P = .96; rMSSD QT): 9.0 +/- 2.4 ms versus 8.7 +/- 2.9 ms, P = .65, and QTc interval 450 +/- 30 ms versus 446 +/- 27 ms, P = .59. At 3 months, the statin group displayed higher HRV SDNN RR): 27.2 +/ 4.9 ms versus 24.4 +/- 2.8 ms in controls, P = .003; rMSSD RR: 24.7 +/- 4.2 ms versus 21.3 +/- 5.6 ms, P = .004, lower QTV SDNN (QT): 5.1 +/- 1.9 ms versus 6.5 +/- 2.1, P = .004; rMSSD (QT): 6.6 +/- 2.8 ms versus 8.8 +/- 3.1 ms, P = .002, and shorter QTc interval 437 +/- 29 ms versus 450 +/- 25 ms, P = .03 than the control group. CONCLUSIONS: Atorvastatin increases HRV, decreases QTV, and shortens QTc interval, and may thereby reduce the risk of arrhythmias in patients with advanced heart failure. PMID- 16360965 TI - Changes in heart rate variability are correlated to hemodynamic improvement with chronic CARVEDILOL therapy in heart failure. AB - BACKGROUND: Reduced heart rate variability (HRV) has been shown to predict mortality in heart failure (CHF). The relationship between improved cardiac function and improvement in HRV has not been previously studied. METHODS AND RESULTS: This was substudy of a randomized, placebo-controlled, double-blinded trial of carvedilol of four months duration. Analysis of HRV was performed on 24 hour Holter monitors obtained at baseline and completion of study. All subjects had symptomatic CHF and an left ventricular ejection fraction (LVEF) <0.35. Study medication was titrated over 1 month to 50 mg/day (< or =75 kg) or 100 mg/day (<75 kg). A total of 17 subjects were randomized to carvedilol and 12 to placebo. Treatment with carvedilol was associated with significant increases in total frequency power, very low frequency power, high frequency power, SDNN, the root mean square of difference of successive RRs, and pNN50. Change in time and frequency domain measures of HRV had a positive correlation with change in LVEF and negative correlation with change in coronary sinus norepinephrine levels. CONCLUSION: Carvedilol therapy in patients with CHF significantly increased HRV. Change in HRV correlates to improved hemodynamics. This suggests that carvedilol therapy partially normalizes autonomic modulation of heart rate in patients with CHF. PMID- 16360966 TI - Sleep and quality of life in stable heart failure. AB - BACKGROUND: Functional performance and mental health are significant quality of life concerns for heart failure (HF) patients. Poor sleep also appears to be common. We examined the extent to which sleep was associated with functional performance and mental health among persons who had stable systolic HF. METHODS AND RESULTS: Sixty-one patients with stable systolic HF wore wrist actigraphs to record nocturnal sleep and daily activity for 3 days while living at home, performed 6-minute walks (6MWT), and completed the Pittsburgh Sleep Quality Index and the Medical Outcomes Study SF-36 questionnaire. Self-reported sleep quality and actigraph-recorded wake time and wake bout time explained 9% to 20% of the variance in the functional performance variables (daytime activity level, 6MWT, self-reported physical function), and mental health, after controlling for age, gender, comorbidity, and New York Heart Association class. Time in bed was negatively associated with functional performance. There were no statistically significant relationships between sleep duration and functional performance. CONCLUSIONS: Self-reported sleep quality and sleep continuity (sleep that is undisturbed by nocturnal awakenings) are associated with functional performance and mental health in stable systolic HF patients. Effective treatment of sleep problems may contribute to improvement in quality of life. PMID- 16360967 TI - B-type natriuretic peptide testing for structural heart disease screening: a general population-based study. AB - BACKGROUND: Several types of structural heart disease are important precursors for congestive heart failure or cardioembolic stroke. We have previously demonstrated that plasma B-type natriuretic peptide (BNP) measurement is useful for detection of structural heart disease in a multiphasic health screening setting. To extend our hypothesis to the general population, the utility of BNP testing for identifying structural heart disease was assessed in a general population and in subgroups divided by sex, age, and presence/absence of risk factors. METHODS AND RESULTS: This cross-sectional cohort study measured plasma BNP concentrations in 993 randomly selected community-dwelling adults (mean age 58 years). All subjects underwent plasma BNP measurement and transthoracic echocardiography. Using prejudged criteria, 41 subjects were diagnosed to have some form of structural heart disease (mild left ventricular systolic dysfunction in 11, valvular heart disease in 9, hypertensive heart disease in 3, hypertrophic cardiomyopathy in 2, ischemic heart disease in 2, lone atrial fibrillation in 14). The utility of BNP testing was evaluated by receiver operating characteristic (ROC) analysis and by cost analysis for detection of 1 case within each subgroup of the cohort. Overall, the sensitivity and specificity of BNP testing for identification of structural heart disease were 61% and 92%, respectively. The area under the ROC curve was 0.77 (95% CI; 0.74-0.79). When sex specific ROC analyses were performed, sensitivity and specificity were 61% and 91% in men, and 50% and 95% in women, respectively. Although the performance of BNP testing on the basis of these figures might be suboptimal, efficacy was improved in subgroups with a high prevalence of heart disease (>8%) such as the cohort aged > or =65 years (men, area under ROC curve = 0.88; cost or = 0.044 mmol/l, 48 h after administration of contrast agent). Eight patients (13%) have demonstrated an increase in their serum creatinine concentration > or = 0.044 mmol/l 30 days after administration of contrast agent: 2/19 (11%) in the control group, 2/21 (10%) in the 600 mg NAC group and 4/20 (20%) the 300 mg NAC group (p = 0.66). The mean volumes of contrast agent used and prehydration given for each of the three groups did not differ significantly (p > 0.83). There was significant improvement in creatinine clearance within each group from baseline to 30 days (p < or = 0.03), but no significant difference between the groups at 48 h and 30 days (p > or = 0.43). Considering the cost of NAC and its administration, we estimate that this would translate to a saving of dollar 26,637 per annum. CONCLUSION: For day stay patients with mild-to-moderate chronic renal impairment undergoing angiography and/or intervention, prehydration alone is less complicated and more cost-effective than a combination of IV NAC (at doses used) and hydration. PMID- 16360995 TI - The time course of recovery following mild thoracic trauma. AB - PURPOSE: The purpose of this study was to determine when patients are able to return to their normal work or home activities. PATIENTS AND METHODS: Eighty cases, 35 females (44%) and 45 males (56%), presenting to our hospital between the years of 2003 and 2004 were prospectively evaluated. There were no hemopneumothorax or associated injury. The patients were randomized into two groups with respect to treatment modalities as the following: non-steroid anti inflammatory (NSAIDs) and Tramadol treatment groups. In the follow-up, visual analog scale (VAS), and vital capacity were conducted on the days 0, 2, 7 and 30 and 6 months. The results were statistically analyzed. RESULTS: The mean age of the group receiving NSAIDs was 55.42 +/- 15.72, and for the group receiving tramadol treatment, it was 55.67 +/- 17.23. VAS result was 7.8 +/- 1.8 in those receiving NSAIDs treatment, while it was 8.2 +/- 1.6 in the other group. The vital capacity was 235.54 +/- 82 ml for the former group, and 232.43 +/- 89.44 for the latter. In their follow-up, particularly by the 7th day, significant improvement of VAS and vital capacity were observed (p < 0.01). Maximum improvement was achieved on the 30th day (p < 0.001). There was no change in the 6th month. CONCLUSION: Patients are able to return to their normal work or home activities after 7 days. Patients experienced steadily decreasing pain and improved vital capacity at each time point after injury. No difference in response was seen between the two medications. A significant improvement is observed by the 7th day after the trauma (p < 0.01). Optimal improvement was achieved on the 30th day (p < 0.001). PMID- 16360996 TI - Minimally invasive surgery for cardiac myxomas using an upper hemi-sternotomy and biatrial septal approach. AB - BACKGROUND: Minimally invasive surgery is pervading all fields of surgery with the principal benefits being: reduced pain, smaller incision, faster recovery, shorter hospital stay and reduced cost. Cardiac surgery is no different. Cardiac myxoma morbidity and mortality depends on early diagnosis, adequate exposure, complete resection, minimal manipulation and unifocal presence. We reviewed our cases of cardiac myxoma excised via an upper hemi-sternotomy with a biatrial septal cardiac approach, to demonstrate this technique as a valid alternative to a full-length median sternotomy. METHODS: From April 1997 to March 1999, one surgeon excised two cardiac myxomas via this minimal technique. Upper hemi sternotomy was midline without transverse sternal transection. Standard aortobicaval cardiopulmonary bypass and myocardial protection were established with complete tumour excision via a biatrial septal approach commenced in the left atrial dome and extended to the right atrial appendage. RESULTS: Inpatient care was prolonged with cardiorespiratory morbidity a consequence of preoperative co-morbidities. Neither patient required return to theatre and hospital mortality was nil. Patients demonstrated reduced sternal wound pain, reduced length of incision, excellent healing and improvement in New York Heart Association functional class with no evidence of recurrence. CONCLUSIONS: Most proposed benefits of minimally invasive surgery were demonstrated in cardiac myxoma application without significant compromise to patient care and recovery using upper hemi-sternotomy and biatrial septal approach. Cardiac myxoma excision via upper hemi-sternotomy and biatrial septal approach can be achieved without compromise to patient care. PMID- 16360997 TI - Giant Left Anterior Descending Artery Aneurysm in a Patient with Behcet's Disease. AB - Coronary artery aneurysm is a rare coronary abnormality nearly half of which originates from atherosclerotic lesions. The major causes of non-atherosclerotic coronary aneurysms include coronary ectasia, Kawasaki disease, Takayasu aortitis, thoracic trauma and complicated angioplasty; however, it has been rarely reported associated with Behcet's disease. PMID- 16360998 TI - Primary pulmonary haemangiopericytoma: a very rare case. AB - Hemangiopericytoma is a mesenchymal neoplasm originating from pericytes--a cell type that surround capillaries. Its primary localization in the lung is extremely rare. A 41-year-old male patient was admitted to our hospital with an abnormal opacity on his chest X-ray. A primary tumor, located in the left upper lobe, was resected, and final diagnosis was hemangiopericytoma of pulmonary origin. The patient has been well for 2 years since the operation with no sign of recurrence or metastasis. A rare case of primary pulmonary tumor has been presented, and possibilities and difficulties in diagnosis as well as therapy are discussed. PMID- 16360999 TI - Obstruction of the right coronary artery ostium by an aortic valve papillary fibroelastoma. AB - Aortic valve papillary fibroelastomas are rare benign tumours which may present with the sequele of embolisation, coronary occlusion or sudden death. We report a case of a 68 year old lady presenting with unstable angina who appeared at coronary angiography to have thrombus in the right coronary ostium. Increasing right territory ischaemia prompted echocardiography, which diagnosed a mass in the right coronary ostium. Urgent surgery was undertaken, demonstrating chronic impaction of an aortic valve fibroelastoma in the right coronary ostium. Aortic valve replacement and coronary bypass grafting to the distal right coronary artery (RCA) was performed. This case illustrates a common cardiology presentation caused by the complication of a rare tumour. PMID- 16361000 TI - National Heart Foundation of Australia and the Cardiac Society of Australia and New Zealand: position statement on lipid management--2005. PMID- 16361001 TI - Progress in the control of hepatitis B infection in the Western Pacific Region. AB - Hepatitis B virus infection is a serious problem globally, and particularly in the Western Pacific Region where the population suffers disproportionately from the infection and its sequelae. By 2001, every immunization programme in the Region had included hepatitis B vaccine in their schedule. However, many challenges remain if every one of the 26 million children born in the 37 countries and areas of the Region each year is to be protected against hepatitis B infection. In 2003, the Regional Committee of the World Health Organization's Western Pacific Region resolved to improve hepatitis B control by making it one of two new pillars for strengthening the Expanded Programme on Immunization. The Committee endorsed the strategies of the Regional Plan to improve hepatitis B control through immunization, reducing chronic HBV infection (chronic carriage rate) to less than 1%, and aiming for coverage of at least 80% of the birth cohort in every district with three doses of hepatitis B vaccine by 2005. To help guide this process, an assessment was made of the progress to date, and is reported in this paper. Coverage data used in this evaluation were not independently verified, and could over-estimate progress made in some countries. Whilst there has indeed been great progress in the Region, a number of national programmes still lack the ability to reach all children with immunization services. Other major issues that need to be addressed are the challenges of delivering a timely birth dose, and for certain countries, the affordability of the vaccine over the short- and long-term. PMID- 16361002 TI - Low titer maternal antibodies can both enhance and suppress B cell responses to a combined live attenuated human rotavirus and VLP-ISCOM vaccine. AB - We investigated effects of low titer (Lo) circulating MatAb on protection and immunogenicity of attenuated (Att) human rotavirus (HRV) priming and 2/6-virus like particle (VLP)-immunostimulating complex (ISCOM) boosting (AttHRV/VLP) or VLP-ISCOM alone vaccines. LoMatAb had both enhancing and suppressing effects on B cell responses, depending on tissue, antibody isotype and vaccine. Differential effects of LoMatAb on IgA responses in different tissues suggest that LoMatAb did not suppress induction of IgA effector and memory B cells but impaired homing of these cells to secondary lymphoid or effector tissues, reducing IgA antibody secreting cells and antibodies at these sites. The AttHRV/VLP vaccine partially overcame LoMatAb suppression, conferred moderate protection against virulent HRV (as measured by reduced viral shedding and diarrhea) and represents a new candidate for rotavirus vaccines for both humans and animals. PMID- 16361003 TI - Hand injuries at a British Military Hospital on operations. AB - The nature of military medical support necessarily changes in the transition from war fighting to the post-conflict phase. This paper examines the activity in the only British Military Hospital serving a multi-national divisional area in Iraq over 2004 during this post-conflict phase. Hand injuries were common and formed a large proportion of the workload seen at the military field hospital on operations. The overwhelming majority of hand trauma resulted in soft tissue injury. There was a clear predisposition to hand trauma for males, manual workers, combat soldiers and engineers/mechanics. While most hand injuries do not require surgical intervention, they impact on the effectiveness of the military population as a result of the large proportion of patients who are placed on restricted duties following hand trauma, 157 of 241 in this study, and the number of soldiers who require aeromedical evacuation for further treatment, 38 of 360 in this study. These injuries require that military surgeons and emergency physicians should be experienced in the initial management of hand trauma and hand trauma should be a core component of their training. The skills of the specialist hand surgeon may be required for definitive management of these injuries at a later stage. PMID- 16361004 TI - Biomechanical evaluation of a new composite bioresorbable screw. AB - A new bioresorbable composite cannulated screw has been developed for small bone fracture fixation. The LG ("Little Grafter") screw is manufactured from Biosteon, which is a composite of poly L-lactic acid and hydroxyapatite. This study aimed to compare interfragmentary compression generated by this new screw with conventional metal screws commonly used in scaphoid fracture fixation. Four small metallic screws were compared with the LG screw, using a bone model produced from rigid polyurethane foam. The screws included the Acutrak, Asnis III, Herbert and Herbert-Whipple screws. The mean maximum compression forces for the LG screw, the Asnis and the Acutrak were comparable (LG 32.3 N, Asnis 32.8 N, Acutrak 38.3 N), whereas those using the Herbert and the Herbert-Whipple screw were significantly lower (Herbert 21.8 N, Herbert-Whipple 19.9 N). The bioresorbable LG screw has been shown to have good compressive properties compared to commonly used small bone fragment compression screws. PMID- 16361005 TI - A growing concern. PMID- 16361006 TI - Multiple sclerosis: advances, excitements, disenchantments. PMID- 16361007 TI - Movement disorders: a step in the right direction. PMID- 16361008 TI - Headache: a good year for research. PMID- 16361009 TI - Neurological infections: clinical advances, global threats. PMID- 16361010 TI - Neuro-oncology: setting new standards of management. PMID- 16361011 TI - Stroke: the catch-up of underinvestigated topics. PMID- 16361012 TI - Dementia: where has 2005 taken us? PMID- 16361013 TI - Neuromuscular disorders: orphan diseases deserve attention. PMID- 16361014 TI - Paediatric neurology: clinical advances with low technology. PMID- 16361015 TI - Dysphagia in acute stroke: a long-awaited trial. PMID- 16361016 TI - Phenobarbital for convulsive epilepsy at primary care level. PMID- 16361017 TI - Antiparkinsonian drugs and visual hallucinations. PMID- 16361019 TI - Neurocysticercosis, mesial temporal lobe epilepsy, and hippocampal sclerosis: an association largely ignored. PMID- 16361020 TI - Behavioural intervention for dysphagia in acute stroke: a randomised controlled trial. AB - BACKGROUND: Swallowing dysfunction after stroke is common, but there is little reliable evidence for how the disorder should be managed. This study compared standard low-intensity and high-intensity behavioural interventions with usual care for dysphagia. METHODS: 306 patients with clinical dysphagia admitted to hospital with acute stroke were randomly assigned to receive usual care (n=102), prescribed by the attending physician; standard low-intensity intervention (n=102), comprising swallowing compensation strategies and diet prescription three times weekly for up to a month; or standard high-intensity intervention and dietary prescription (n=102), at least daily for up to a month. The primary outcome measure was survival free of an abnormal diet at 6 months. Analysis was done by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00257764. FINDINGS: 60 patients died and three patients were lost to follow up before the 6-month analysis. Of patients randomly allocated usual care, 56% (57/102) survived at 6 months free of an abnormal diet compared with 67% (136/204) allocated standard swallowing therapy (relative risk 1.19, 95% CI 0.98 1.45). Standard swallowing therapy was associated with a non-significant trend toward a reduction in death (0.80, 0.5-1.3), institutionalisation (0.69, 0.4 1.1), and dependency (1.05, 0.8-1.3); a significant reduction in swallowing related medical complications (0.73, 0.6-0.9), chest infection (0.56, 0.4-0.8), and death or institutionalisation (0.73, 0.55-0.97); and a significant rise in the proportion of patients regaining swallowing function (1.41, 1.03-1.94) by 6 months. Compared with usual care and low-intensity therapy, high-intensity therapy was associated with an increased proportion of patients who returned to a normal diet (p=0.04) and recovered swallowing (p=0.02) by 6 months. INTERPRETATION: These data show a consistent trend towards more favourable outcomes in dysphagic stroke patients who are assigned a standard programme of early behavioural swallowing intervention, including active therapeutic approaches and dietary modification. PMID- 16361021 TI - Efficacy and safety of dexanabinol in severe traumatic brain injury: results of a phase III randomised, placebo-controlled, clinical trial. AB - BACKGROUND: Traumatic brain injury is a major cause of death and disability. We sought to assess the safety and efficacy of dexanabinol, a synthetic cannabinoid analogue devoid of psychotropic activity, in severe traumatic brain injury. METHODS: 861 patients with severe traumatic brain injury admitted to 86 specialist centres from 15 countries were included in a multi-centre, placebo controlled, phase III trial. Patients were randomised to receive a single intravenous 150 mg dose of dexanabinol or placebo within 6 h of injury. The primary outcome was the extended Glasgow outcome scale assessed at 6 months, with the point of dichotomisation into unfavourable versus favourable outcome differentiated by baseline prognostic risk. Prespecified subgroup analyses were defined by injury severity, recruitment rate, and time to dosing. Secondary analysis included control of intracranial pressure and quality of life. Analysis were prespecified in the protocol and the statistical analysis plan. This study is registered with ClinicalTrials.gov, number NCT00129857. FINDINGS: 846 patients were included in the efficacy analysis. The extended Glasgow outcome scale at 6 months did not differ between groups; 215 (50%) patients in the dexanabinol group and 214 (51%) patients in the placebo group had an unfavourable outcome (odds ratio for a favourable response 1.04; 95% CI 0.79-1.36). Improvements in the control of intracranial pressure or quality of life were not recorded and subgroup analysis showed no indication of differential treatment effects. Dexanabinol was not associated with hepatic, renal, or cardiac toxic effects. INTERPRETATION: Dexanabinol is safe, but is not efficacious in the treatment of traumatic brain injury. PMID- 16361022 TI - Efficacy assessment of phenobarbital in epilepsy: a large community-based intervention trial in rural China. AB - BACKGROUND: Many people with epilepsy need not experience further seizures if the diagnosis and treatment are correct. Most epilepsy patients have convulsions, which are usually fairly easy to diagnose. This study tested a model for treatment of people with convulsive forms of epilepsy at primary health-care level in rural areas of China. METHODS: Patients with convulsive epilepsy were identified at primary care level and provided with phenobarbital monotherapy. Local physicians, who were provided with special training, carried out screening, treatment, and follow-up. A local neurologist confirmed the diagnoses. Efficacy was assessed from the percentage reduction in seizure frequency from baseline and the retention of patients on treatment. FINDINGS: The study enrolled 2455 patients. In 68% of patients who completed 12 months' treatment, seizure frequency was decreased by at least 50%, and a third of patients were seizure free. 72% of patients who completed 24 months' treatment had reduction of seizure frequency of at least 50% and a quarter of patients remained seizure free. Probability of retention was 0.84 at 1 year, and 0.76 at 2 years. Medication was well tolerated and reported adverse events were mild; only 32 patients (1%) discontinued medication because of side-effects. INTERPRETATION: This pragmatic study confirmed that this simple protocol was suitable for the treatment of convulsive forms of epilepsy in rural areas of China. Physicians with basic training could treat epilepsy patients with phenobarbital, with beneficial effects for most patients with convulsive seizures. Few cognitive or behavioural adverse events were noted, but formal psychometric testing was not done. PMID- 16361023 TI - Mechanisms of brain injury after intracerebral haemorrhage. AB - The past decade has resulted in a rapid increase in knowledge of mechanisms underlying brain injury induced by intracerebral haemorrhage (ICH). Animal studies have suggested roles for clot-derived factors and the initial physical trauma and mass effect as a result of haemorrhage. The coagulation cascade (especially thrombin), haemoglobin breakdown products, and inflammation all play a part in ICH-induced injury and could provide new therapeutic targets. Human imaging has shown that many ICH continue to expand after the initial ictus. Rebleeding soon after the initial haemorrhage is common and forms the basis of a current clinical trial using factor VIIa to prevent rebleeding. However, questions about mechanisms of injuries remain. There are conflicting data on the role of ischaemia in ICH and there is uncertainty over the role of clot removal in ICH therapy. The next decade should bring further information about the underlying mechanisms of ICH-induced brain injury and new therapeutic interventions for this severe form of stroke. This review addresses our current understanding of the mechanisms underlying ICH-induced brain injury. PMID- 16361024 TI - Risk of dementia in diabetes mellitus: a systematic review. AB - The relation between diabetes and major types of dementia is controversial. This systematic review examines the incidence of dementia in people with diabetes mellitus. We identified 14 eligible longitudinal population-based studies of variable methodological quality. The incidence of "any dementia" was higher in individuals with diabetes than in those without diabetes in seven of ten studies reporting this aggregate outcome. This high risk included both Alzheimer's disease and vascular dementia (eight of 13 studies and six of nine studies respectively). Detailed data on modulating and mediating effects of glycaemic control, microvascular complications, and comorbidity (eg, hypertension and stroke) were generally absent. The findings of mechanistic studies suggest that vascular disease and alterations in glucose, insulin, and amyloid metabolism underlie the pathophysiology, but which of these mechanisms are clinically relevant is unclear. Further high quality studies need to be initiated, with objective diabetes assessment, together with reliable methods to establish the contribution of vascular disease and other comorbidity to dementia. PMID- 16361025 TI - The diagnosis of Parkinson's disease. AB - The correct diagnosis of Parkinson's disease is important for prognostic and therapeutic reasons and is essential for clinical research. Investigations of the diagnostic accuracy for the disease and other forms of parkinsonism in community based samples of patients taking antiparkinsonian medication confirmed a diagnosis of parkinsonism in only 74% of patients and clinically probable Parkinson's disease in 53% of patients. Clinicopathological studies based on brain bank material from the UK and Canada have shown that clinicians diagnose the disease incorrectly in about 25% of patients. In these studies, the most common reasons for misdiagnosis were presence of essential tremor, vascular parkinsonism, and atypical parkinsonian syndromes. Infrequent diagnostic errors included Alzheimer's disease, dementia with Lewy bodies, and drug-induced parkinsonism. Increasing knowledge of the heterogeneous clinical presentation of the various parkinsonisms has resulted in improved diagnostic accuracy of the various parkinsonian syndromes in specialised movement-disorder units. Also genetic testing and various other ancillary tests, such as olfactory testing, MRI, and dopamine-transporter single-photon-emission computed-tomography imaging, help with clinical diagnostic decisions. PMID- 16361026 TI - A life-course approach to the aetiology of late-onset dementias. AB - Substantial progress has been made in the understanding of the neurobiology of dementias, but comprehensive causal models are not available. Genetic and environmental factors probably interact to determine vulnerability to the dementias. The life-course approach to age-related diseases, when systematically applied to the dementias, provides opportunities to identify the nature and timing of environmental contributions. We discuss the relevance of the fetal origins of adult disease hypothesis to the dementias. Associations between the dementias (most often described as Alzheimer's disease) and ischaemic heart disease, obesity, hypertension, hyperlipidaemia, and non-insulin-dependent diabetes mellitus are set against associations between dementias and childhood intelligence, low educational attainments, low socioeconomic status, occupation, and lifetime dietary history. Biological mechanisms that explain how fetal development might influence the risk of adult disease may be relevant to many age related diseases including the dementias and, possibly, to the biology of ageing. PMID- 16361027 TI - A 32-year-old man with relapsing-progressive brainstem symptoms. PMID- 16361028 TI - Metals distribution in soils around the cement factory in southern Jordan. AB - Thirty one soil samples were collected from south Jordan around the cement factory in Qadissiya area. The samples were obtained at two depths, 0-10 cm and 10-20 cm and were analyzed by atomic absorption spectrophotometery for Pb, Zn, Cd, Fe, Cu and Cr. Physicochemical factors believed to affect their mobility of metals in soil of the study area were examined such as; pH, TOM, CaCO3, CEC and conductivity. The relatively high concentrations of lead, zinc and cadmium in the soil samples of the investigated area were related to anthropogenic sources such as cement industry, agriculture activities and traffic emissions. It was found that the lead, zinc and cadmium have the highest level in area close to the cement factory, while the concentration of chromium was low. This study indicate that all of the metals are concentrated on the surface soil, and decreased in the lower part of the soil, this due to reflects their mobility and physical properties of soil and its alkaline pH values. The use of factor analysis showed that anthropogenic activities seem to be the responsible source of pollution for metals in urban soils. PMID- 16361029 TI - Gender, cancer experience and internet use: a comparative keyword analysis of interviews and online cancer support groups. AB - A new method, comparative keyword analysis, is used to compare the language of men and women with cancer in 97 research interviews and two popular internet based support groups for people with cancer. The method is suited to the conjoint qualitative and quantitative analysis of differences between large bodies of text, an alternative to the 'code and retrieval' approach used in much thematic analysis of qualitative materials. Web forums are a rich source of data about illness experience and gender differences. Marked differences in the performance of gender are evident. These differences follow linguistic and other behavioural patterns (such as social network differences) established in other contexts. Men with prostate cancer indicate in research interviews that they are more likely to seek information on the internet; women with breast cancer that they are more likely to seek social and emotional support. Men's concerns cluster around treatment information, medical personnel and procedures. Their experience of disease is more localised on particular areas of the body, while women's experience is more holistic. Women's forum postings orientate much more towards the exchange of emotional support, including concern with the impact of illness on a wide range of other people. Women's use of superlatives as well as words referring to feelings indicate their enactment of greater emotional expressivity. Web forums are platforms for an intensification of men's knowledge gathering activities. Web forums, though actually quite publicly visible, appear to be subjectively experienced by both sexes as relatively private places for the exchange of intimate personal information. The 'privacy' of the breast cancer forum facilitated interactions found in other studies to be characteristic of women's friendship groups. PMID- 16361031 TI - Childhood obesity--a public health crisis. PMID- 16361032 TI - Communicating the ocular and systemic complications of obesity to patients. AB - BACKGROUND: Obesity has become a growing worldwide epidemic that is associated with serious medical complications. Many clinicians and patients are aware of the systemic complications of obesity, but there are few providers and patients who are aware of the ocular manifestations of obesity. METHODS: A review of the current medical literature was performed. Ocular and systemic manifestations associated with obesity are described, and a methodology for educating the optometric patient is presented. RESULTS: There is significant morbidity and mortality associated with obesity, which is causing serious consequences to patients and an enormous economic strain on the health care system. Optometrists have a tremendous opportunity and obligation to educate their patients on the potential sight-threatening complications of obesity and encourage them to decrease their caloric intake and increase their physical activity to help alleviate this individual and collective burden. CONCLUSIONS: Optometrists have a duty to educate and counsel their overweight and obese patients on the devastating ocular complications that could manifest as a result of obesity. PMID- 16361033 TI - Recurrent corneal erosion (RCE) secondary to lattice dystrophy in a patient with acquired immune deficiency syndrome (AIDS). AB - BACKGROUND: We present a case of an Acquired Immune Deficiency Syndrome (AIDS) patient with recurrent erosion (RCE) secondary to lattice corneal dystrophy. As a human immunodeficiency virus (HIV)-infected patient becomes more immunocompromised, the ocular surface defense mechanisms may be compromised. Lattice dystrophy, RCE, and modifying approaches to the management of RCE in an HIV-positive or AIDS patient are reviewed. CASE REPORT: A 49-year-old man presented with RCE. His ocular history included lattice corneal dystrophy OU, recurrent corneal erosion O.S., and herpes simplex virus keratitis O.S. Systemic history included hepatitis C and HIV infection with a diagnosis of AIDS with secondary Pneumocystis carinii pneumonia. Viral load was 35533 HIV-RNA (ribonucleic acid) molecules/ml of plasma and CD4 lymphocyte count 99 cells/mm3. Acuities were O.D. 20/80 and O.S. 20/50. The abrasion was treated with cycloplegia and bacitracin/polymyxin B ointment q.i.d. O.S. and it resolved in 3 days. CONCLUSION: Management of lattice dystrophy with secondary RCE in an AIDS patient requires that the clinician be familiar with the patient's immune status. As the CD4 count declines and the viral load increases, the patient is at higher risk for opportunistic anterior segment infections. Clinicians need to monitor these patients closely for potential complicating ocular sequelae of AIDS such as herpes zoster ophthalmicus, herpes simplex keratitis, fungal/bacterial keratitis, and keratoconjunctivitis sicca. Our patient responded well to antibiotic therapy and cycloplegia. The importance of daily monitoring of immunocompromised patients is emphasized. PMID- 16361034 TI - Descemet's membrane detachment after cataract extraction. AB - BACKGROUND: Descemet's membrane detachment is a rare but potentially serious complication of intraocular surgery, most commonly cataract extraction. Small Descemet's membrane detachments typically resolve with topical medical therapy; however, larger detachments require surgical intervention. The most common surgical procedure is a gas-fluid exchange with 20% sulfur hexafluoride (SF6) or 14% perfluropropane (C3F8), which is typically performed at the biomicroscope and is nontoxic to the endothelium. CASE: A 77-year-old man underwent phacoemulsification with a posterior chamber lens implantation by clear corneal incision in the left eye. Visual acuity was 20/40 in the left eye 1 month postoperatively with persistent corneal edema. Visual acuity continued to deteriorate to counting fingers (CF), despite the use of topical hyperosmotics and steroids. At 2 months, a scrolled Descemet's membrane detachment was present from the incision site to the central cornea causing extensive edema. Anterior chamber injection of 14% C3F8 successfully reattached Descemet's membrane. Six weeks after surgery, the gas bubble had resolved, the central cornea was clear, and a curvilinear scar was present from 5:00 to 11:00. Visual acuity remained at CF owing to anterior capsular fibrosis. Anterior YAG capsulotomy improved the vision to 20/40. CONCLUSION: Descemet's membrane detachment can have a devastating effect on vision. Timely management is imperative to preserve vision. This case shows the successful repair of Descemet's membrane detachment with 14% C3F8 after unsuccessful topical treatment. PMID- 16361035 TI - Characterization of cadmium uptake in human intestinal crypt cells HIEC in relation to inorganic metal speciation. AB - Cadmium (Cd) uptake was studied under inorganic exposure conditions in normal human intestinal crypt cells HIEC. The uptake time course of 0.3 microM Cd in a serum-free chloride medium was analyzed according to a first order equation with rapid initial (U0) and maximal (Umax) accumulation values of 14.1+/ 1.4pmol/mgprotein and 41.4+/-2.0 pmol/mgprotein, respectively. The presence of a 300-fold excess of unlabeled Cd dramatically decreased tracer uptake, showing the involvement of specific mechanism(s) of transport. Our speciation studies revealed the preferential uptake of the free ion Cd2+, but also suggested that CdCln(2-n) species may contribute to Cd accumulation. Specific mechanisms of transport of very high and similar affinity (Km approximately 5 microM) have been characterized under both chloride and nitrate exposure conditions, but a two-fold higher capacity (Vmax) was estimated in the nitrate medium used to increase [Cd2+] over chlorocomplex formation. A clear inhibition of 109Cd uptake was observed at external acidic pH under both exposure media. An La-inhibitible 46% increase in 109Cd uptake was obtained in nominally Ca-free nitrate medium, whereas Zn provided additional inhibition. These results show different kinetic parameters for Cd uptake as a function of inorganic metal speciation. Cd2+ uptake would not involve the H+-coupled symport NRAMP2 but would be related instead to the Ca and/or Zn pathways. Because proliferative crypt cells play a critical role in the renewal process of the entire intestinal epithelium, studies on the impact of Cd on HIEC cell functions clearly deserve further investigation. PMID- 16361036 TI - Pancreaticoduodenectomy following endoscopic placement of an expandable metal biliary stent. PMID- 16361037 TI - Esophageal pseudodiverticulosis. PMID- 16361038 TI - Pyogenic liver abscess after colonoscopy in a patient with ulcerative colitis. PMID- 16361039 TI - GIH clinical research update: 2004-2005. AB - This review highlights areas of clinical research in gastroenterology and hepatology that were published during the last year and were summarized during the most recent American Gastroenterological Association Plenary Session. The topics include a comparison of the risk of recurrent bleeding in patients taking clopidogrel versus aspirin plus a proton pump inhibitor, the introduction of rifaximin for the treatment of traveler's diarrhea, and the results of an oral vaccine for cholera tested in a high endemic area where there is also a high prevalence of human immunodeficiency virus infection. In inflammatory bowel disease, the impact of a biomarker of inflammation, C-reactive protein, to the response to a new biologic therapy is identified as potentially important because it might facilitate the selection of patients for these treatments. Results of device, endoscopic, and surgical treatment of obesity are reviewed, including the evidence of significant impact of surgery-induced weight loss on comorbid diseases. In the field of cancer, colonoscopic screening results in more polyps detected, down-staging of cancers identified, and improved cancer survival. A new familial syndrome associated with a serrated adenoma/carcinoma phenotype and variability in microsatellite instability is described. A controlled study demonstrates that a urine-derived substance, ulinastatin, reduces the risk of post-endoscopic retrograde cholangiopancreatography pancreatitis. Hepatic stellate cells are involved in the fibrogenesis associated with nonalcoholic fatty liver disease. These areas of clinical research demonstrate the breadth of significant advances that will impact on the clinical practice of gastroenterology and hepatology. PMID- 16361040 TI - Advances in gastrointestinal pharmacotherapy. AB - The medical management of patients with gastrointestinal diseases is advancing rapidly. At a recent symposium held during Digestive Disease Week in Chicago in May of 2005, specific attention was given to the future prospects for medical management of 3 common gastrointestinal disease areas: antisecretory therapy, chronic hepatitis C, and inflammatory bowel disease. Antisecretory approaches include drug combinations including a proton pump inhibitor, potassium competitive acid blockers, and antigastrin agents. The latter two classes are still experimental, but the former combinations have potential to enhance the highly effective agents currently available. The focus of treatment advances in chronic hepatitis C in the immediate future is the discovery of more effective treatment regimens for nonresponders to prior therapy, who are becoming the largest group of patients seeking treatment of hepatitis C. The combination of peginterferon with ribavirin results in 6%-15% sustained virologic response rates in patients who were prior nonresponders to standard interferon plus ribavirin. Newer strategies to eradicate hepatitis C virus infection using different interferons, such as interferon alfacon-1 or higher doses of peginterferon, or long-term maintenance peginterferon, are undergoing study and show promise based on data from preliminary studies. Several immunomodulators have promise in inflammatory bowel disease, although the risk-benefit ratio and costs of therapy require evaluation. Nevertheless, the success of new biologics such as anti TNFalpha agents augurs well for effective future therapies. PMID- 16361041 TI - Gastroduodenal disease, Helicobacter pylori, and genetic polymorphisms. AB - Over the past 20 years, there has been marked progress in our understanding of the role of genetic and environmental factors in the etiology of gastroduodenal disease. Helicobacter pylori infection now is recognized to be the most important environmental factor for both noncardia gastric cancer and peptic ulcer disease. The likelihood of the infection resulting in significant disease depends on genetic polymorphisms influencing the virulence of the organism. However, the specific pattern of disease induced by the infection is determined to a great extent by genetic polymorphisms in the host that govern the local gastric immune response elicited. Genetic factors also are important in the treatment of gastroduodenal diseases. Polymorphisms of host CYP2C19 influence the pharmacokinetics and clinical efficacy of proton pump inhibitor therapy. PMID- 16361042 TI - Management of refractory ascites. PMID- 16361043 TI - Microarrays and prediction of interferon response in chronic hepatitis C: not yet ready for prime time. PMID- 16361044 TI - The bulge, booze, and the liver. PMID- 16361045 TI - Eosinophilic esophagitis: a 10-year experience in 381 children. AB - BACKGROUND & AIMS: Eosinophilic esophagitis (EoE) is a disorder characterized by a severe, isolated eosinophilic infiltration of the esophagus unresponsive to aggressive acid blockade but responsive to the removal of dietary antigens. We present information relating to our 10-year experience in children diagnosed with EoE. METHODS: We conducted a retrospective study between January 1, 1994, and January 1, 2004, to evaluate all patients diagnosed with EoE. Clinical symptoms, demographic data, endoscopic findings, and the results of various treatment regimens were collected and evaluated. RESULTS: A total of 381 patients (66% male, age 9.1 +/- 3.1 years) were diagnosed with EoE: 312 presented with symptoms of gastroesophageal reflux; 69 presented with dysphagia. Endoscopically, 68% of patients had a visually abnormal esophagus; 32% had a normal-appearing esophagus despite a severe histologic esophageal eosinophilia. The average number of esophageal eosinophils (per 400 x high power field) proximally and distally were 23.3 +/- 10.5 and 38.7 +/- 13.3, respectively. Corticosteroids significantly improved clinical symptoms and esophageal histology; however, upon their withdrawal, the symptoms and esophageal eosinophilia recurred. Dietary restriction or complete dietary elimination using an amino acid-based formula significantly improved both the clinical symptoms and esophageal histology in 75 and 172 patients, respectively. CONCLUSIONS: Medications such as corticosteroids are effective; however, upon withdrawal, EoE recurs. The removal of dietary antigens significantly improved clinical symptoms and esophageal histology in 98% of patients. PMID- 16361046 TI - Inappropriate use of intravenous pantoprazole: extent of the problem and successful solutions. AB - BACKGROUND & AIMS: Indications for intravenous proton pump inhibitors (IV PPI) include upper gastrointestinal bleeding (UGIB) from peptic ulcer disease with high-risk stigmata and patients receiving nothing by mouth (NPO). The objectives were to assess the extent of errors in indications for IV PPI use and to determine whether multidisciplinary interventions could improve IV PPI use and costs. METHODS: Part 1: Patients prescribed IV PPI during a period of 4 months were divided into 2 settings, UGIB or non-UGIB. The setting-specific appropriateness of the IV PPI indication and dosing regimen was determined. Part 2: Patients prescribed IV PPI before and after multidisciplinary interventions (educating physicians, a computerized dose template, pharmacists altering IV PPI orders in non-UGIB patients who were not NPO, and recommending a GI consult when a continuous infusion was ordered) were studied. Incidence of prescribing errors, IV PPI costs, and potential confounders were compared. RESULTS: Part 1: Only 50% of UGIB (n = 145) patients received IV PPI for an appropriate indication. Both indication and dosing regimen were appropriate in 21%. In the non-UGIB group (n = 95), 33% were truly NPO; 51% had a correct dosing frequency. Part 2: The postintervention (n = 105) group (vs the preintervention group, n = 113) showed a significant absolute reduction in the degree of inappropriate indication in the UGIB (26%; 95% confidence interval [CI], 10%-42%; P < .0001) and in the non-UGIB (41%; 95% CI, 24%-58%; P < .0001) subgroups. However, a greater improvement in underspending than overspending meant that overall costs were unchanged. CONCLUSIONS: IV PPI was frequently prescribed inappropriately and incorrectly; simple maneuvers resulted in reductions in errors. PMID- 16361047 TI - Effect of systemic corticosteroid therapy on risk for intra-abdominal or pelvic abscess in non-operated Crohn's disease. AB - BACKGROUND & AIMS: Systemic corticosteroid therapy increases risk of postoperative sepsis in Crohn's disease. This study investigates its effect on risk for sepsis in non-operated patients. METHODS: A retrospective case-control study was performed in 432 patients with Crohn's disease (the 94% of our database for whom adequate documentation could be retrieved). Two analyses were performed. The first tested the hypothesis that patients with perforating Crohn's disease (n = 86) were more likely to develop intra-abdominal or pelvic abscess (n = 29) if they had received systemic corticosteroids during the previous 3 months. The second analysis, confined to interventions since 1998, tested the hypothesis that corticosteroid therapy was more common during the 3 months before presentation with intra-abdominal or pelvic abscess (n = 12) than during the 3 months after presentation with a relapse of nonperforating disease (n = 24 consecutive patients). In both analyses adjustment was made for any other significant variable. RESULTS: Systemic corticosteroid therapy was associated with an adjusted odds ratio (OR) for intra-abdominal or pelvic abscess of 9.03 (95% confidence interval [CI], 2.40-33.98) in patients with perforating Crohn's disease. Patients receiving prednisolone > or = 20 mg per day had an OR of 2.81 (95% CI, 0.99-7.99) for abscess compared with those receiving a lower dose. In patients with relapsed active disease, corticosteroid therapy was associated with an unadjusted OR of 9.31 (95% CI, 1.03-83.91) for intra-abdominal or pelvic abscess. Neither smoking nor azathioprine usage was associated with increased risk for abscess. CONCLUSIONS: Systemic corticosteroid therapy for Crohn's disease is associated with increased risk for intra-abdominal or pelvic abscess. PMID- 16361048 TI - Magnetic resonance imaging compared with ileocolonoscopy in evaluating disease severity in Crohn's disease. AB - BACKGROUND & AIMS: Abdominal magnetic resonance imaging (MRI) has shown promising results in the detection of Crohn's disease (CD)-related lesions. The purpose of this study was to assess the value of MRI in measuring disease activity in CD patients in comparison with ileocolonoscopy. METHODS: Thirty-one patients undergoing ileocolonoscopy because of suspicion of relapsing CD underwent MRI with water as intraluminal contrast medium. At endoscopy, disease severity was graded (4-point scale), and Crohn's Disease Endoscopic Index of Severity (CDEIS) was determined. Two radiologists independently interpreted the MRI scans. Radiologic grading (4-point scale) was compared with endoscopic grading of disease severity and CDEIS (overall, for all segments). Wall thickness and enhancement were compared with CDEIS. Patient experience and preference were determined. RESULTS: In, respectively, 14 and 14 patients (radiologist 1) and 16 and 11 patients (radiologist 2) an exact match or 1 level of difference in grading was scored with the endoscopist. Correlation between severity rated at MRI and CDEIS was moderate to strong with r = 0.61 (P < .001) for observer 1 and r = 0.63 (P < .001) for observer 2. Per segment, best correlation was seen in the terminal ileum (r = 0.63; P < .001, for both observers). Wall thickness correlated moderately to strongly with CDEIS (r = 0.57, P < .001 and r = 0.50, P < .001 for observers 1 and 2), whereas enhancement correlated weakly to moderately (r = 0.45, P < .001 and r = 0.42, P < .001). Patients experienced more pain during endoscopy, and all patients except 2 preferred MRI to endoscopy. CONCLUSION: MRI can correctly identify disease severity in patients with CD and is a patient-friendly alternative to ileocolonoscopy. PMID- 16361049 TI - Expandable metal biliary stents before pancreaticoduodenectomy for pancreatic cancer: a Monte-Carlo decision analysis. AB - BACKGROUND & AIMS: Endoscopic placement of plastic or self-expandable metal biliary stents (SEMS) relieves obstructive jaundice from pancreatic cancer. Short length, distally placed SEMS do not preclude subsequent pancreaticoduodenectomy. We sought to determine whether SEMS placement in patients whose surgical status is uncertain is cost-effective for management of obstructive jaundice. METHODS: A Markov model was constructed to evaluate costs and outcomes associated with endoscopic biliary stenting for obstructive jaundice. Strategies evaluated were: (1) initial plastic stent with plastic stents for subsequent occlusions in nonsurgical candidates after staging (plastic followed-up by [f/u] plastic), (2) initial plastic with subsequent SEMS (plastic f/u metal), (3) initial short length SEMS with subsequent plastic (metal f/u plastic), and (4) initial short length SEMS with subsequent expandable metal stent (metal f/u metal). Published stent occlusion rates, ERCP complication rates and outcomes, cholangitis rates and outcomes, pancreatic cancer mortality rates, and Whipple complication rates were used. Costs were based on 2004 Medicare standard allowable charges and were accrued until all patients reached an absorbing health state (death or pancreaticoduodenectomy) or 24 cycles (24 mo) ended. RESULTS: Average costs per patient from Monte Carlo simulation were: (1) metal f/u metal, $19,935; (2) plastic f/u metal, 20,157 dollars; (3) metal f/u plastic, 20,871 dollars; and (4) plastic f/u plastic, 20,878 dollars. For initial plastic stents to be preferred over short-length metal stents, 70% or more of pancreatic cancers would need to be potentially resectable by pancreaticoduodenectomy. CONCLUSIONS: In patients undergoing ERCP before definitive cancer staging, short-length SEMS is the preferred initial cost-minimizing strategy. PMID- 16361050 TI - Comparison of early endoscopic ultrasonography and endoscopic retrograde cholangiopancreatography in the management of acute biliary pancreatitis: a prospective randomized study. AB - BACKGROUND & AIMS: The role and potential benefits of endoscopic ultrasonography (EUS) in the management of acute biliary pancreatitis have not been documented. We report a large prospective randomized study comparing early EUS and endoscopic retrograde cholangiopancreatography (ERCP) in the management of these patients. METHODS: A prospective randomized study was performed on 140 patients with acute pancreatitis suspected to have a biliary cause. The patients were randomized to have EUS (n = 70) or ERCP (n = 70) within 24 hours from admission. In the EUS group, when EUS detected choledocholithiasis, therapeutic ERCP was performed during the same endoscopy session. In the ERCP group, diagnostic ERCP was performed, followed by therapeutic endoscopy when choledocholithiasis was detected. RESULTS: Examination of the biliary tree by EUS was successful in all patients in the EUS group, whereas cannulation of the common duct during ERCP was unsuccessful in 10 patients (14%) in the ERCP group (P = .001). Combined percutaneous ultrasonography and ERCP missed detection of cholelithiasis in 6 patients in the ERCP group. The overall morbidity rate was 7% in the EUS group, and that in the ERCP group was 14% (P = .172). The hospital stay and mortality rates were comparable in both groups. CONCLUSIONS: In selected patients with acute biliary pancreatitis, EUS could safely replace diagnostic ERCP in the management for selecting patients with choledocholithiasis for therapeutic ERCP with a higher successful examination rate, a higher sensitivity in the detection of cholelithiasis, and a comparable morbidity rate. PMID- 16361051 TI - Prophylactic balloon-occluded retrograde transvenous obliteration for gastric varices in compensated cirrhosis. AB - BACKGROUND & AIMS: Because gastric variceal bleeding is associated with a high mortality rate, its prophylaxis is expected to improve survival. We investigated the effectiveness of balloon-occluded retrograde transvenous obliteration (B-RTO) for the treatment of gastric fundal varices. METHODS: A prospective nonrandomized study was conducted. Prophylactic B-RTO was performed in 17 patients (B-RTO group), whereas the remaining 17 patients received no specific treatment (control). The nonbleeding rate, cumulative survival rate, and prognostic values of the patients were assessed. RESULTS: The respective nonbleeding rates at 1, 3, and 5 years were 100%, 100%, and 83% in the B-RTO group and 81%, 59%, and 39% in the control. The respective cumulative survival rates at 1, 3, and 5 years were 94%, 85%, and 39% in the B-RTO group and 71%, 41%, and 22% in the control. Both the nonbleeding rate and the cumulative survival rate of the B-RTO group were significantly higher than those of the control (P = .01 and .04, respectively). B RTO was determined by multivariate analysis to be a significant factor for low bleeding rate (relative risk, 0.06; 95% confidence interval [CI], 0.004-0.79), whereas B-RTO (0.11; 95% CI, 0.03-0.44) and Child-Pugh class A (0.10; 95% CI, 0.03-0.39) were the significant factors for a low mortality rate, and the presence of hepatocellular carcinoma (5.68; 95% CI, 1.49-21.7) was the significant factor for a high mortality rate. CONCLUSIONS: Prophylactic B-RTO is effective in preventing gastric variceal rupture and consequently improves patient survival. PMID- 16361052 TI - Pretreatment prediction of interferon-alfa efficacy in chronic hepatitis C patients. AB - BACKGROUND & AIMS: Interferon has been used widely to treat patients with chronic hepatitis C infections. Prediction of interferon efficacy before treatment has been performed mainly by using viral information, such as viral load and genotype. This information has allowed the successful prediction of sustained responders (SR) and non-SRs, which includes transient responders (TR) and nonresponders (NR). In the current study we examined whether liver messenger RNA expression profiles also can be used to predict interferon efficacy. METHODS: RNA was isolated from 69 liver biopsy samples from patients receiving interferon monotherapy and was analyzed on a complementary DNA microarray. Of these 69 samples, 31 were used to develop an algorithm for predicting interferon efficacy, and 38 were used to validate the precision of the algorithm. We also applied our methodology to the prediction of the efficacy of interferon/ribavirin combination therapy using an additional 56 biopsy samples. RESULTS: Our microarray analysis combined with the algorithm was 94% successful at predicting SR/TR and NR patients. A validation study confirmed that this algorithm can predict interferon efficacy with 95% accuracy and a P value of less than .00001. Similarly, we obtained a 93% prediction efficacy and a P value of less than .0001 for patients receiving combination therapy. CONCLUSIONS: By using only host data from the complementary DNA microarray we are able to successfully predict SR/TR and NR patients for interferon therapy. Therefore, this technique can help determine the appropriate treatment for hepatitis C patients. PMID- 16361053 TI - Joint effects of body weight and alcohol on elevated serum alanine aminotransferase in the United States population. AB - BACKGROUND & AIMS: Both alcohol and obesity are associated with hepatic steatosis. However, little is known about whether the toxicity of alcohol to the liver is influenced by adiposity. We examined the relationship of alcohol drinking and binge drinking with abnormal serum aminotransferase activity in normal weight, overweight, and obese persons in a national, population-based study. METHODS: Data were analyzed from 13,580 adult participants in the third US National Health and Nutrition Examination Survey, 1988-1994, after excluding participants with hepatitis B or C or iron overload. Abnormal aminotransferase levels were defined by using sex-specific cutoffs for ALT and AST. Analyses were adjusted for other liver injury risk factors. RESULTS: The prevalence of abnormal aminotransferase activity was elevated with consumption of >2 drinks per day or with overweight and obesity. In multivariate analysis, there was no association of alcohol intake with a higher prevalence of elevated aminotransferase levels among normal weight persons. In contrast, among overweight persons, consumption of >2 drinks per day increased the risk of elevated aminotransferase levels, and among the obese, > or = 1 drink per day was associated with a higher risk. Results were similar with elevated ALT alone as the outcome. With elevated AST alone as the outcome, intake of >2 drinks per day increased the risk, even among normal weight persons. Binge drinking was associated with aminotransferase elevation among obese consumers of 1-2 drinks per day. CONCLUSIONS: In this large, national, population-based study, overweight and obesity increased the risk of alcohol-related abnormal aminotransferase activity. PMID- 16361054 TI - Effect of interferon-gamma on hepatic fibrosis in chronic hepatitis B virus infection. PMID- 16361055 TI - Determination of the optimum conditions in the removal of Bomaplex Red CR-L dye from the textile wastewater using O3, H2O2, HCO3- and PAC. AB - Bomaplex Red CR-L textile dye was used in the experimental studies. Taguchi method was applied to determine optimum conditions in the removal of dye from synthetic textile wastewater. After the parameters were determined to remove Bomaplex Red CR-L dye from synthetic textile wastewater, the experimental studies were realized. The chosen experimental parameters and their ranges: HCO3- (mM), 0 39; temperature (degrees C), 18-70; ozone-air flow rate (l min-1), 5-15; the dye concentration (ppm), 200-600; particulate activated carbon (PAC) (g), 0-1.5; H2O2 (mM), 0-0056; pH, 3-12; and treatment time (min), 10-30, respectively. An orthogonal array L18 (2(1)x3(7)) for experimental plan and the smaller the better performance statistics formula were selected to define optimum conditions. The optimum conditions were found to be as follows: HCO3- (mM), 0; temperature (degrees C), 70; ozone-air flow rate (l min-1), 10; the dye concentration (ppm), 200; particulate activated carbon (PAC) (g), 1; H2O2 (mM), 0.056; pH, 12; and time (min), 20. Under these optimum conditions, it was determined that the Bomaplex Red CR-L removal efficiency from textile wastewater was 99%. PMID- 16361056 TI - Characterization and leachability of electric arc furnace dust made from remelting of stainless steel. AB - Electric arc furnace dust (EAFD) is a toxic waste product made in the remelting of scrap steel. The results of a Toxicity Characteristic Leaching Procedure (TCLP) conducted on a sample of EAFD originating from the remelting of stainless steel scrap showed that the total Cr and Cr (VI) liquor concentrations (9.7 and 6.1 mg/L, respectively) exceeded the Toxicity Characteristic Regulatory Level (TCRL). The EAFD showed a complex heterogeneous mineralogy with spinel minerals group predominance. A sequential extractions method has permitted the determination of the amount of available metals (potentially mobile component) from the EAFD as follows: Cr (3%), Ni (6%), Pb (49%) and Zn (40%). Solubility controls on Cr, Pb, Zn and Ni were identified in the EAFD. This means that the Cr, Pb, Zn and Ni concentrations in solution were controlled by the solubility of some phases from EAFD. The concentrations of Ni and Zn, which are metals not regulated by TCRL were below 0.41 and 1.3 mg/L, respectively. The solubility control on Pb was sufficient to decrease its concentration (<0.24 mg/L) to a level below the TCRL. However, the control on Cr was not sufficient to decrease its concentration (between 117 and 331 mg/L) to below the TCRL. PMID- 16361057 TI - Chemical denitrification of water by zero-valent magnesium powder. AB - A laboratory-scale study was conducted in batch mode to investigate the feasibility of using zero-valent magnesium (Mg(0)), for removal of nitrate from aqueous solution. Reaction pH, dose of Mg(0), initial nitrate concentration and temperature were considered variable parameters during the study. Strong acidic condition enhanced nitrate reduction and in absence of external proton addition, reaction pH increased rapidly above ten and insignificant nitrate removal (7-16%) was achieved. At Mg(0):NO(3)(-)-N molar ratio of 5.8 and controlled reaction pH of 2, 84% denitrification efficiency was achieved (initial NO(3)(-)-N 50 mg/L) under ambient temperature and pressure and total nitrogen removal was 70% with 3.2% and 10% conversion of initial NO(3)(-)-N to NO(2)(-)-N and NH(4)(+)-N, respectively. The reaction was first order with respect to nitrate concentration. Nitrate removal rate decreased with solution pH and increased linearly with Mg(0) dose. Nitrate removal was coupled with 96-100% removal of dissolved oxygen and 85 90% generation of soluble Mg(2+) ion. An activation energy (E(a)) of nitrate reduction over the temperature range of 10-50 degrees C was observed as 17.7 kJ mol(-1). PMID- 16361058 TI - Exploiting structural principles to design cyclin-dependent kinase inhibitors. AB - Although cyclin-dependent kinases (CDKs) have been extensively targeted in anti cancer drug design, no CDK inhibitor has yet been approved for use in cancer therapy. While this may in part be because inhibitors clinically evaluated to date have not demonstrated clean inhibition of a single CDK, another contributing factor is an apparent latent functional redundancy in the CDK cell-cycle regulatory system. This further complicates the already challenging goal of targeting CDKs, since it implies that a therapeutically useful inhibitor will have to selectively inhibit more than one CDK family member among the complement of cellular proteins. Despite these difficulties, achieving an appropriate profile of CDK inhibition may yet be possible using ATP-competitive inhibitors, thanks to advances in computational and experimental methods of drug design. However, as an alternative to ATP-competitive inhibitors, inhibitors that interfere with a CDK-specific protein:protein interaction, such as that which occurs at the recruitment site found on several cyclins, may offer a route to a therapeutically useful inhibitory profile. PMID- 16361059 TI - Unpleasant odors increase pain processing in a patient with neuropathic pain: psychophysical and fMRI investigation. AB - A 49-year old man with neuropathic pain in his right elbow, wrist and digits III V of his hand reported that certain odors increased his pain by superimposing an electric shock-like pain to his already existing pain. Psychophysical testing revealed that the best predictor of pain exacerbation was odor unpleasantness. Functional magnetic resonance imaging (fMRI) showed increased activation following an unpleasant odor in pain related areas, including the thalamus, amygdala, insular and anterior cingulate cortices, with similar trends in primary somatosensory cortex hand/arm area. The increased pain and associated neural activations in response to unpleasant odors may be related to the phenomenon of synesthesia, to a rewiring of olfactory pathways onto pain pathways mimicking synesthesia or, to activation of the sympathetic nervous system. PMID- 16361060 TI - The effects of noxious heat, auditory stimulation, a cognitive task, and time on task on pain perception and performance accuracy in healthy volunteers: a new experimental model. AB - The effects of cognitive and competing sensory processing tasks on pain perception and as a function of time are only partially understood. To study these effects, we compared the simultaneous effects of noxious heat stimulation (HS), auditory stimulation (AS) (sinusoidally modulated speech-like signal, SMSLS), and a cognitive task (CT) (rate change detection of the SMSLS) on pain perception and task performance over repeated experimental runs. Sixty healthy paid volunteers were randomly assigned to four groups, one exposed to AS while performing the CT, one to HS (46 degrees C/6 min), one to AS and HS, and one to AS and HS while performing the CT. Each group performed the experimental run four times, each run for 6 min. Immediately after each run, the subjects rated pain intensity using a VAS (0-100). Two-way RM-ANOVA for analyzing pain intensities among the three heat pain groups demonstrated significant differences of VAS ratings (F(2,179) = 4.57, P = 0.019), being highest in the HS group (55 +/- 0.7SEM), followed by the AS+HS (39 +/- 6.8) and AS + HS + CT (33 +/- 0.7) groups. Post-hoc analyses revealed that group HS differed significantly from group AS + HS + CT and from group AS + HS (P < 0.05, SNK), whereas group AS+HS did not differ significantly from group AS + HS + CT. Neither pain rating, nor rate of errors on the CT varied significantly across runs. These findings point to a significant influence of competing passive sensory processing on pain perception, with the cognitive task not necessarily adding to the perception of pain. Advantages and shortcomings of the present experimental model for future pain studies are discussed. PMID- 16361061 TI - Relative virulences of a drug-resistant and a drug-sensitive strain of Eimeria acervulina, a coccidium of chickens. AB - The virulence of a field strain of the chicken coccidian parasite Eimeria acervulina (Boreham I), dually resistant to the chemically unrelated anticoccidial agents decoquinate and clopidol, was compared with that of a drug sensitive laboratory strain (Ongar) of the same species. Following a single heavy infection (prevented from recycling), both strains exhibited pathogenic effects typical of their species, viz., pathognomonic lesions, adverse effects on body weight gain and feed conversion ratio (FCR), but no mortality. One week after infection, chicks infected with either strain had a statistically significantly worse weight gain than the uninfected control; the Boreham I strain produced more oocysts, and caused slightly more severe duodenal lesions and poorer FCRs than the Ongar strain (all those effects being non-significant). After 3 weeks, there were no significant differences between any cumulative effects of either strain, nor any differences from the uninfected control. However, from 2 to 3 weeks after infection, chicks infected with either strain had a greater feed consumption and growth rate than uninfected chicks. When chicks reared on solid floors were given lighter infections of either strain, which were allowed subsequently to recycle naturally, there were no consistent reductions in weight gains, but feed consumption was higher than that of uninfected chicks. Whatever, the mode of infection, there were no significant differences between the weights of infected and uninfected chicks after 3 weeks, but the FCR of infected chicks was usually poorer than that of uninfected chicks. The difference between the virulences of the Boreham I and Ongar strains was not greater than that between various drug resistant strains or between various sensitive strains of several Eimeria species recorded in the literature. It is therefore concluded that there was no difference between the virulences of the two strains of E. acervulina that could be attributed to the drug-resistance of one of them. PMID- 16361062 TI - Optical recording from respiratory pattern generator of fetal mouse brainstem reveals a distributed network. AB - Unfailing respiration depends on neural mechanisms already present in mammals before birth. Experiments were made to determine how inspiratory and expiratory neurons are grouped in the brainstem of fetal mice. A further aim was to assess whether rhythmicity arises from a single pacemaker or is generated by multiple sites in the brainstem. To measure neuronal firing, a fluorescent calcium indicator dye was applied to embryonic central nervous systems isolated from mice. While respiratory commands were monitored electrically from third to fifth cervical ventral roots, activity was measured optically over areas containing groups of respiratory neurones, or single neurones, along the medulla from the facial nucleus to the pre-Botzinger complex. Large optical signals allowed recordings to be made during individual respiratory cycles. Inspiratory and expiratory neurones were intermingled. A novel finding was that bursts of activity arose in a discrete area intermittently, occurring during some breaths, but failing in others. Raised CO2 partial pressure or lowered pH increased the frequency of respiration; neurons then fired reliably with every cycle. Movies of activity revealed patterns of activation of inspiratory and expiratory neurones during successive respiratory cycles; there was no evidence for waves spreading systematically from region to region. Our results suggest that firing of neurons in immature respiratory circuits is a stochastic process, and that the rhythm does not depend on a single pacemaker. Respiratory circuits in fetal mouse brainstem appear to possess a high safety factor for generating rhythmicity, which may or may not persist as development proceeds. PMID- 16361063 TI - Differential, strain-specific cellular and subcellular distribution of multidrug transporters in murine choroid plexus and blood-brain barrier. AB - Multidrug transporters of the ATP-binding cassette family play an important role in regulating drug distribution and efflux in the brain, owing to their selective distribution in microvessels and choroid plexus. Their expression pattern and cellular distribution remain controversial, in part due to technical difficulties in localizing these membrane proteins in closely associated cells, such as endothelial cells and astrocytic end-feet at the blood-brain barrier. Here, we used high-resolution immunofluorescence staining with cell-type specific markers to investigate the distribution of major ATP-binding cassette transporters in mouse brain. We report that four ATP-binding cassette transporters, Mdr1, Mrp1, Mrp2 and Mrp5 can be detected in brain endothelial cells, forming three distinct layers, with Mdr1 and Mrp5 being located on the luminal side, Mrp1 on the abluminal (basal) side, and Mrp2 in between. Mrp3 and Mdr3 were undetectable. In choroid plexus, only Mrp1, Mrp2 and Mrp3 were detected, again with a differential distribution. Mrp1 was targeted basolaterally in epithelial cells, Mrp2 was restricted to endothelial cells, and Mrp3 was co-localized with zonula occludens 1 at tight junctions. Analysis of Mdr1a(0/0) and Mrp1(0/0) mice, generated in the FVB strain, revealed no major alteration in expression of the remaining transporters. An unexpected strain difference was unraveled, with wildtype FVB mice selectively lacking Mrp2 protein in brain, but not liver. In conclusion, these results indicate that ATP-binding cassette transporters provide multiple penetration barriers in the blood-brain barrier and choroid plexus, with a selective cellular and subcellular distribution, emphasizing their potential role for drug resistance in neurological disorders, such as epilepsy. PMID- 16361064 TI - Altered discharges of spinal wide dynamic range neurons and down-regulation of glutamate transporter expression in rats with paclitaxel-induced hyperalgesia. AB - Changes in the signaling of wide dynamic range neurons and the expression of glutamate transporters in the lumbar spinal dorsal horn of rats with Taxol induced hyperalgesia are detailed in this report. Deep spinal lamina neurons have significantly increased spontaneous activity and after-discharges to noxious mechanical stimuli, increased responses to both skin heating and cooling, and increased after-discharges and abnormal windup to transcutaneous electrical stimuli. The expression of glutamate transporter proteins in the dorsal horn is decreased at the time point corresponding to the physiological changes. These results suggest a state of increased excitability develops in spinal pain signaling neurons as a consequence of decreased glutamate clearance. These changes in dorsal horn neurobiology likely in turn contribute to the hyper responsiveness to sensory stimuli seen in animals treated with Taxol and may play a role in the pain seen in cancer patients receiving Taxol. PMID- 16361065 TI - Afferent connections of the subparafascicular area in rat. AB - The subparafascicular nucleus and the subparafascicular area are the major sites of synthesis of the recently discovered neuropeptide, tuberoinfundibular peptide of 39 residues (TIP39). Better knowledge of the neuronal inputs to the subparafascicular area and nucleus will facilitate investigation of the functions of TIP39. Thus, we have injected the retrograde tracer cholera toxin B subunit into the rostral, middle, and caudal parts of the rat subparafascicular nucleus. We report that the afferent projections to the subparafascicular nucleus and area include the medial prefrontal, insular, and ectorhinal cortex, the subiculum, the lateral septum, the anterior amygdaloid area, the medial amygdaloid nucleus, the caudal paralaminar area of the thalamus, the lateral preoptic area, the anterior, ventromedial, and posterior hypothalamic nuclei, the dorsal premamillary nucleus, the zona incerta and Forel's fields, the periaqueductal gray, the deep layers of the superior colliculus, cortical layers of the inferior colliculus, the cuneiform nucleus, the medial paralemniscal nucleus, and the parabrachial nuclei. Most of these regions project to all parts of the subparafascicular nucleus. However, the magnocellular subparafascicular neurons, which occupy the middle part of the subparafascicular nucleus, may not receive projections from the medial prefrontal and insular cortex, the medial amygdaloid nucleus, the lateral preoptic area, and the parabrachial nuclei. In addition, double labeling of cholera toxin B subunit and TIP39 revealed a remarkable similarity between input regions of the subparafascicular area and the brain TIP39 system. Neurons within regions that contain TIP39 cell bodies as well as regions that contain TIP39 fibers project to the subparafascicular area. Overall, the afferent connections of the subparafascicular nucleus and area suggest its involvement in central reproductive, visceral, nociceptive, and auditory regulation. PMID- 16361066 TI - Strain-dependent expression of signaling proteins in the mouse hippocampus. AB - Individual mouse strains may differ significantly in terms of behavior and cognitive function. Hippocampal gene expression profiling on several mouse strains has been carried out and points toward substantial strain-specific variation of more than 200 genes including components of major signaling pathways involved in neuronal information storage. Strain-specific hippocampal protein expression, however, has not been investigated yet. A proteomic approach based on two-dimensional gel electrophoresis coupled with mass spectrometry has been chosen to address this question by determining strain-dependent expression of signaling proteins in hippocampi of four inbred and one outbred mouse strain. Forty-six spots corresponding to 37 different signaling proteins have been analyzed and quantified. Statistical analysis revealed strain-dependent expression of serine/threonine protein phosphatase 1, serine/threonine protein phosphatase 2A, large GTP binding protein OPA1, guanine nucleotide-binding protein beta, putative GTP-binding protein Ran, receptor of activated protein kinase C1, WASP-family protein member 1, voltage-dependent anion channel 2 and 14 3-3 protein gamma. Differential expression of signaling proteins in the hippocampus may contribute to the molecular understanding of strain-dependent behavioral and cognitive performance. Moreover, these data highlight the importance of the genetic background for the analysis of signaling pathways in the hippocampus in wild-type mice as well as in gene-targeting experiments. PMID- 16361067 TI - A direct projection from superior colliculus to substantia nigra pars compacta in the cat. AB - Dopaminergic neurons exhibit a short-latency, phasic response to unexpected, biologically salient stimuli. The midbrain superior colliculus also is sensitive to such stimuli, exhibits sensory responses with latencies reliably less than those of dopaminergic neurons, and, in rat, has been shown to send direct projections to regions of the substantia nigra and ventral tegmental area containing dopaminergic neurons (e.g. pars compacta). Recent electrophysiological and electrochemical evidence also suggests that tectonigral connections may be critical for relaying short-latency (<100 ms) visual information to midbrain dopaminergic neurons. By investigating the tectonigral projection in the cat, the present study sought to establish whether this pathway is a specialization of the rodent, or whether it may be a more general feature of mammalian neuroanatomy. Anterogradely and retrogradely transported anatomical tracers were injected into the superior colliculus and substantia nigra pars compacta, respectively, of adult cats. In the anterograde experiments, abundant fibers and terminals labeled with either biotinylated dextran amine or Phaseolus vulgaris leucoagglutinin were seen in close association with tyrosine hydroxylase-positive (dopaminergic) somata and processes in substantia nigra pars compacta and the ventral tegmental area. In the retrograde experiments, injections of biotinylated dextran amine into substantia nigra produced significant retrograde labeling of tectonigral neurons of origin in the intermediate and deep layers of the ipsilateral superior colliculus. Approximately half of these biotinylated dextran amine-labeled neurons were, in each case, shown to be immunopositive for the calcium binding proteins, parvalbumin or calbindin. Significantly, virtually no retrogradely labeled neurons were found either in the superficial layers of the superior colliculus or among the large tecto-reticulospinal output neurons. Taken in conjunction with recent data in the rat, the results of this study suggest that the tectonigral projection may be a common feature of mammalian midbrain architecture. As such, it may represent an additional route by which short latency sensory information can influence basal ganglia function. PMID- 16361068 TI - Evolutionary conservation and mutational spectrum of BMPR2 gene. AB - A variety of mutations in the bone morphogenetic protein receptor type 2 (BMPR2) have been identified in patients with pulmonary arterial hypertension. In this study, using our BMPR2 mutation database and BMPR-II protein sequences from eight distantly related species, we defined the relationship among evolutionary conservation, mutation frequency and mutation distribution. As a whole, BMPR2 is evolving slower than the average for mammalian protein-encoding genes. As expected, the kinase domain is evolving more slowly than the extracellular ligand binding and C-terminal domains. A detailed map of evolutionary conservation shows that there are repeating peaks and valleys within the C-terminal domain, representing higher and lower evolutionary conservation. We observed a strong correlation between evolutionary conservation and the distribution of mutations along the gene. All except two, of the nineteen missense mutations occur in absolutely conserved amino acids among the vertebrate homologs. In addition, we identified six mutational hotspots (P<0.05) by comparing the observed distribution of mutations to the pattern expected from a random multinomial distribution. Furthermore, analysis of the sequence environment surrounding the mutations revealed a specific pattern of mutagenesis. Over 22% of all single base paired substitutions and 30% of all deletions and insertions are situated within tandem or non-tandem direct repeats of at least 5-bp and may be explained by slipped-mispairing model of mutagenesis. Also, over 59% of single base-paired substitutions versus 20% of deletions and insertions are located in perfect palindromic sequences that could produce "hairpin-loop" secondary structures with relatively high thermodynamic stability under physiological conditions. In addition, 3.7% of single base-paired substitutions versus 30% of deletions and insertions are located either within or in close proximity to the Krawczak and Cooper consensus sequence (TG A/G A/G G/T A/C). Further study of the mechanism of mutagenesis in BMPR2 may help identify other potentially mutable sites and differentiate between deleterious mutations and harmless polymorphic variants. PMID- 16361070 TI - Development of a cell culture assay for the quantitative determination of vaccination-induced antibodies in rabbit sera against Clostridium perfringens epsilon toxin and Clostridium novyi alpha toxin. AB - Cell culture assays are possible alternatives to replace in vivo neutralization tests currently required for potency testing of clostridial vaccines. Cell culture assays based on the MDCK cell line and the Vero cell line which are sensitive to the Clostridium (C.) perfringens type D epsilon toxin and Clostridium novyi type B alpha toxin, respectively, were developed, and the test conditions were standardized. The antibody titres of vaccinated rabbits measured in vitro were compared with the results of current test procedures recommended by European Pharmacopoeia. The correlation coefficients calculated were significant for all sera tested. The cell culture assays proved to be sensitive, specific, reproducible and reliable. Therefore, these cell culture assays could be suitable in vitro alternatives to the in vivo mouse neutralization experiments required for potency tests of clostridial vaccines, but further validation studies are necessary. PMID- 16361071 TI - Contact sensitisation and allergic contact dermatitis: immunobiological mechanisms. AB - The dose-response relationships of the human immune system can be defined using the induction and elicitation of lymphocyte mediated allergic reactions to experimental contact sensitisers such as dinitrochlorobenzene (DNCB). Five groups of healthy volunteers received a sensitising dose of DNCB applied to a 3 cm diameter circle on the volar forearm. Doses applied were 62.5 microg, 125 microg, 250 microg, 500 microg and 1,000 microg. Four weeks later a concentration series of 3.125 microg, 6.25 microg, 12.5 microg and 25 microg was applied to the upper inner arm on 1cm paper discs which were removed after 6h. Forty-eight hours later the responses were scored clinically and thickness measured with callipers. The proportion of people reacting to the challenge doses showed a sigmoid log-dose response curve, 100% reacting to 500 microg. The responses to challenge also showed a log-dose-reponse. As sensitising dose increased so more people were sensitised to a proportionately greater degree. These dose-response relationships reflect the effects of increasing the concentration of sensitiser on a fixed area. The effect was examined of keeping the concentration per sq cm constant but of varying the total area. When 35.4 microg/cm(2), which sensitised 80% of people when applied to a circle 3 cm diameter (area 7.1cm(2)), was applied on a 1.5 cm diameter circle or 4.5 cm diameter, there were no differences in the proportions sensitised or their degree of reactivity. This was clearly on the plateau for the sensitising effect. However, when the same concentration per cm(2) was applied on a 3mm diameter area much weaker sensitisation was obtained. This shows the concentration of sensitiser per unit area is the critical determinant of whether sensitisation occurs, whereas the total dose may be varied over a wide range, but if the concentration per unit area is constant there is no effect on sensitising potency. In other words few Langerhans cells presenting many antigen molecules per cell is a much more potent sensitising stimulus than the same number of molecules presented by many Langerhans cells, each presenting few molecules. These observations clearly have important implications across the whole field of risk assessment for induction of contact sensitivity. PMID- 16361069 TI - Molecular pathways mediating mechanical signaling in bone. AB - Bone tissue has the capacity to adapt to its functional environment such that its morphology is "optimized" for the mechanical demand. The adaptive nature of the skeleton poses an interesting set of biological questions (e.g., how does bone sense mechanical signals, what cells are the sensing system, what are the mechanical signals that drive the system, what receptors are responsible for transducing the mechanical signal, what are the molecular responses to the mechanical stimuli). Studies of the characteristics of the mechanical environment at the cellular level, the forces that bone cells recognize, and the integrated cellular responses are providing new information at an accelerating speed. This review first considers the mechanical factors that are generated by loading in the skeleton, including strain, stress and pressure. Mechanosensitive cells placed to recognize these forces in the skeleton, osteoblasts, osteoclasts, osteocytes and cells of the vasculature are reviewed. The identity of the mechanoreceptor(s) is approached, with consideration of ion channels, integrins, connexins, the lipid membrane including caveolar and non-caveolar lipid rafts and the possibility that altering cell shape at the membrane or cytoskeleton alters integral signaling protein associations. The distal intracellular signaling systems on-line after the mechanoreceptor is activated are reviewed, including those emanating from G-proteins (e.g., intracellular calcium shifts), MAPKs, and nitric oxide. The ability to harness mechanical signals to improve bone health through devices and exercise is broached. Increased appreciation of the importance of the mechanical environment in regulating and determining the structural efficacy of the skeleton makes this an exciting time for further exploration of this area. PMID- 16361072 TI - Growth inhibition, drug load, and degradation studies of gelatin/methotrexate conjugates. AB - Macromolecular gelatin-methotrexate conjugates have potential therapeutic advantages over the free drug. Conjugates with MTX:gelatin molar ratios (MR) ranging from 1:1 to 27:1 were examined for cell growth inhibition, stability, degradation, and methotrexate (MTX) release. Conjugate growth inhibition was less than that of free MTX whose IC(50) value of 1.3 x 10(-8) M was about 10-fold less. Cell uptake of fluorescein labeled gelatin (145 kD) was observed by 24-30 h. Higher MR conjugates produced less growth inhibition, measurably greater stability at pH 7.4 based on MTX release, and had less gelatin degradation in the conjugate by the lysosomal enzyme Cathepsin B (Cat B) compared to low MR conjugates. Cat B conjugate degradation was greater at the in vitro lysosomal pH of 4.8 than the intra-tumor pH of 6.5. The presence of Cat B did not meaningfully affect MTX release, but less MTX was released at pH 4.8 than pH 6.5. The maximum MTX release was a relatively low 7% after 72 h at pH 6.5 for the low MR conjugate. Low molecular weight conjugate fragments were also produced and were also influenced by pH and MR. Reduced growth inhibition by high MR conjugates may be due to a hindered enzymatic degradation in the lysosomes. A strong peptide conjugate bond at lysosomal pH and a 24-30 h delayed gelatin uptake may contribute to reduced growth inhibition of the conjugate compared to free MTX. MTX release under these in vitro conditions occurs by aqueous hydrolysis, not by Cat B cleavage of the conjugate bond. PMID- 16361073 TI - Ethyl acetate extract of Patrinia scabiosaefolia downregulates anti-apoptotic Bcl 2/Bcl-X(L) expression, and induces apoptosis in human breast carcinoma MCF-7 cells independent of caspase-9 activation. AB - Patrinia scabiosaefolia Fisch. is a Chinese medicinal herb used traditionally for treating intestinal carbuncle. Although Patrinia scabiosaefolia has also been suggested for cancer therapy, there has not been any scientific evidence supporting this application. In this study, a panel of human cancer cells, including breast carcinoma MCF-7; hepatocellular carcinoma HepG2; skin melanoma A375; lung carcinoma A549 and prostate adenocarcinoma PC-3, were treated in vitro with ethyl acetate extract of Patrinia scabiosaefolia (EAE-PS) for 48 h. Results from MTT study showed that MCF-7 was the most responsive (IC50 = 112.3 microg/ml) while PC-3 was the most resistant (IC50 = 348.7 microg/ml) one to cell growth inhibition. DNA flow cytometry demonstrated that EAE-PS induced apoptosis in the resistant MCF-7 cells by 14.5-fold of the control level after 36 h of treatment. Immunoblot studies further illustrated that although EAE-PS downregulated the anti-apoptotic Bcl-2/Bcl-X(L) expression in breast cancer cells, the induced apoptosis could not be prevented by the caspase-9 inhibitor (Z-LEHD-FMK). All these results suggest that EAE-PS retards MCF-7 cell growth by activating the caspase-independent mitochondrial cell death pathway. Results from this study support future research and development of the bioactive ingredients from Patrinia scabiosaefolia as anticancer agents, especially against those apoptosis resistant cancers with deregulated Bcl-2/Bcl-X(L) expression. PMID- 16361074 TI - Some mathematical problems in the DNA identification of victims in the 2004 tsunami and similar mass fatalities. AB - DNA is a major and essential identification tool for mass fatality incidents including the hundreds of thousands of victims of the 2004 Indian Ocean tsunami. Mathematical complications characteristic of this sort of mass fatality include prevalence of related victims, the many races represented among the victims, and various identification modalities in tandem with DNA. Four mathematical problems of interest are discussed in this paper. (1) Other quantifiable factors (i.e. geography) can be formally accounted for by including a likelihood ratio that can be thought of as reducing the "effective number of victims." (2) When a victim is found and tentatively identified as V, but then it comes to light that the victim has a relative W who is also missing, confidence in the identity is depressed. To account for the existence of W, increment the effective number of victims by the likelihood ratio supporting W as the identity of the victim. (3) When several apparently related victims are found, their mutual identities should be calculated simultaneously. Compared to one-at-a-time, serial identifications, this is both logical and may lead to much more confidence in the identities. (4) Although there may be many different population groups represented among the missing, it is generally sufficient to consider population statistics for only a few of them in deciding whether to declare an identification. PMID- 16361075 TI - Impairment of water maze behaviour with ageing is counteracted by maze learning earlier in life but not by physical exercise, food restriction or housing conditions. AB - Spatial learning and memory decline with ageing in humans as well as rats. We examined the influence of different interventions on male Sprague Dawley rats with respect to learning ability and memory at the age of 5, 10 and 18 months. The intervention and control groups were: (RW) voluntary exercise in running wheels, (PW) sedentary, food restricted (by about 25%) to keep them at pair weight with RW, (S1) sedentary, fed ad libitum, (TM) forced training in a treadmill, and, (S4) sedentary, fed ad libitum. The animals in all groups were housed individually except those in group S4, which were housed four in each cage. The ability of learning and memory was determined in the Morris water maze. The results showed a significantly better learning ability when young in comparison with their ability when having grown older. At the age of 18 months, the performance was significantly better in the subgroups which had received training also at the age of 10 months compared to the subgroups receiving their first training. None of the various interventions had any significant effect on these functions. Repeated training seems to be the best intervention with respect to retaining learning ability and memory. PMID- 16361076 TI - Quantitative analysis of PPARdelta mRNA by real-time RT-PCR in 86 rectal cancer tissues. AB - AIM: The purpose of this study is to clarify the expression change of PPARdelta gene in human colorectal cancer tissues. METHODS: Applying real-time RT-PCR, we quantified PPARdelta mRNA in a series of 86 tissues from excised primary rectal cancers. In each case, accompanying normal mucosa was collected for comparison. RESULTS: Among the 86 rectal cancer tissues, 48 cases showed PPARdelta overexpression: 39 tumours gave an expression level 1.5-5 times, five tumours 10 20 times, and four tumours more than 20 times relative to normal mucosa. However, the general level of PPARdelta mRNA in rectal cancer tissues is not statistically different from normal mucosa. CONCLUSIONS: The expression of PPARdelta gene in rectal cancers is not statistically different from normal mucosa. PMID- 16361077 TI - [Anaesthesia for cerebral radiotherapy: about 35 repeated intubations for a same 30 months old child]. PMID- 16361078 TI - Urotensin-II expression in the mouse spinal cord. AB - Urotensin-II (UII), a 12 amino acid peptide, was discovered in the teleost fish neurosecretory cells located in the caudal portion of the spinal cord and which project to a neurohemal gland called the urophysis. The distribution of UII and of its prepro-UII mRNA is not limited to fish and was found for example in the rat spinal cord. In view of the potential interest of obtaining transgenic mice, we have therefore characterized the distribution of mouse pro-UII mRNA and UII immunoreactivity, by in situ hybridization and immunohistochemistry, respectively, in the mouse spinal cord. A population of UII-like immunoreactive cell bodies was located in the ventral horn of the different segments. These cells displayed all the features of motoneurons, as confirmed by a double immunohistochemical labelling showing the co-occurrence of UII and vesicular acetylcholine transporter, and by electron microscope immunocytochemistry. Retrograde labelling of motoneurons innervating the bulbocavernosus penile muscle showed that some of them contained UII. In situ hybridization histochemistry revealed that pro-UII mRNA was located in some ventral horn neuronal perikarya. The pro-UII mRNA-containing cell bodies possessed the same motoneuron characteristics, confirming the results of the immunohistochemical studies and showing that the gene of mouse UII is expressed in a subpopulation of motoneurons in the spinal cord. Our results support the assumption that UII peptide characterized as endocrine in fish is also expressed within mammalian motoneurons. PMID- 16361079 TI - Predictors of stress in mothers and fathers of children with fragile X syndrome. AB - This study examined parental and family stress and functioning where there is a child with fragile X syndrome. Mothers and fathers in 40 families were asked about their child with fragile X syndrome, family supports, their psychological stress, the marital relationship, and their family stress. Results indicate parents were well adjusted in terms of their levels of psychological stress and in their marital relationships, however, parents reported high levels of family stress. Mothers and fathers were found to experience similar levels of stress and to report similar levels of satisfaction with supports. Stress was predicted by different variables in mothers and fathers, suggesting that different processes underlie their experiences. The strongest predictor of maternal stress was the level of marital satisfaction while the strongest predictor of paternal stress was the level of the child's adaptive skills. PMID- 16361080 TI - Fuzzy c-means clustering with spatial information for image segmentation. AB - A conventional FCM algorithm does not fully utilize the spatial information in the image. In this paper, we present a fuzzy c-means (FCM) algorithm that incorporates spatial information into the membership function for clustering. The spatial function is the summation of the membership function in the neighborhood of each pixel under consideration. The advantages of the new method are the following: (1) it yields regions more homogeneous than those of other methods, (2) it reduces the spurious blobs, (3) it removes noisy spots, and (4) it is less sensitive to noise than other techniques. This technique is a powerful method for noisy image segmentation and works for both single and multiple-feature data with spatial information. PMID- 16361081 TI - TGF-beta regulates differentially the proliferation of fetal and adult human skin fibroblasts via the activation of PKA and the autocrine action of FGF-2. AB - Transforming growth factor-beta (TGF-beta) is a potent regulator of cell proliferation; interestingly its action is clearly cell type-dependent. In particular, it inhibits epithelial and endothelial cells' proliferation, while its action on many mesenchymal cells has been reported to be stimulatory. In this direction, we have recently shown that TGF-beta regulates the proliferation of normal human skin fibroblasts according to their developmental origin: i.e. it inhibits fetal fibroblasts, while it stimulates the proliferation of adult ones. Here, we present evidence on the mechanisms underlying this differential action. Concerning fetal fibroblasts, we have found that TGF-beta activates Protein Kinase A (PKA) and induces the expression of the cyclin-dependent kinase inhibitors (CKIs) p21(CIP1/WAF1) and p15(INK4B). Moreover, the specific PKA inhibitor H-89 blocks the induction of both CKIs and annuls the TGF-beta-mediated inhibitory effect, indicating the central role of PKA in this process. In contrast, in adult cells no PKA activation is observed. Moreover, TGF-beta stimulates cell proliferation by activating the MEK-ERK pathway, as the MEK inhibitor PD98059 blocks this effect. A specific neutralizing antibody against Fibroblast Growth Factor-2 (FGF-2) inhibits both ERK activation and the mitogenic activity of TGF-beta, indicating that the latter establishes an autocrine loop, via FGF-2, leading to cell proliferation. This loop requires FGF receptor-1 (FGFR 1), as its down-regulation by siRNA approach prevents TGF-beta from stimulating ERK-1/2 activation and DNA synthesis. In conclusion, the differential proliferative response of fetal and adult normal human skin fibroblasts to TGF beta is regulated by distinct signaling pathways and furthermore it may provide information on the bimodal effect of this factor on cell proliferation, in general. PMID- 16361082 TI - Distribution and functional activity of P-glycoprotein and multidrug resistance associated proteins in human brain microvascular endothelial cells in hippocampal sclerosis. AB - Multidrug resistance protein, also referred as P-glycoprotein (P-gp, MDR1; ABCB1) and multidrug resistance-associated protein (MRP) 1 (ABCC1) and 2 (ABCC2) are, thus far, candidates to cause antiepileptic drug (AED) resistance epilepsy. In this study, we investigated P-gp, MRP1 and MRP2 expression, localization and functional activity on cryosections and isolated human brain-derived microvascular endothelial cells (HBMEC) from epileptic patients (HBMEC-EPI) with hippocampal sclerosis (HS), as compared with HBMEC isolated from normal brain cortex (HBMEC-CTR). We examined the expression and distribution of three transporters, P-gp, MRP1 and MRP2 on two major parts of the resected tissue, the hippocampus and the parahippocampal gyrus (Gph). P-gp showed diffuse expression not only in endothelium but also by parenchymal cells in both the hippocampus and the Gph. MRP1 labeling was observed in parenchymal cells in the Gph. By contrast, MRP2 was mainly found in endothelium of the hippocampus. P-gp and MRP1 expression in the Gph was relatively high in the patient with long-term seizure history. Quantitative RT-PCR analysis of HBMEC revealed that MDR1, MRP1 as well as MRP5 (ABCC5) and MRP6 (ABCC6) were overexpressed in HBMEC-EPI at the mRNA level. HBMEC from both normal and epilepsy groups displayed protein expression of P-gp, whereas MRP1 and MRP2 were seen only in HBMEC-EPI. Accordingly, it is of particular interest that MRP functional activities were observed in HBMEC-EPI, but not in HBMEC-CTR. Our results suggest that complex MDR expression changes not only in the hippocampus but in the Gph may play a role in AED pharmacoresistance in intractable epilepsy patients with mesial temporal lobe epilepsy (MTLE) by altering the permeability of AEDs across the blood-brain barrier (BBB). PMID- 16361083 TI - Expression of dihydrodiol dehydrogenase plays important roles in apoptosis- and drug-resistance of A431 squamous cell carcinoma. AB - BACKGROUND: Dihydrodiol dehydrogenase (DDH) is a member of the aldo-keto reductases superfamily which may be involved in normal detoxification process of environmental mutagenic hazards like polycyclic aromatic hydrocarbons (PAH). Previous clinical studies have demonstrated the over-expression of DDH in various types of cancers, including cutaneous squamous cell carcinoma (SCC), and its correlation with tumor progression and grave prognosis. OBJECTIVE: To investigate possible mechanisms for DDH's correlation with tumor progression and unfavorable prognosis. METHODS: DDH expression in SCC A431 cell line was examined by quantitative real-time PCR and immunoblotting. RNA interference (RNAi) by transduction with retroviral vector containing DDH-targeting small interfering RNA was employed to inhibit DDH expression by A431 cells. With DDH expression inhibited or not, sensitivity of A431 cells to UVB-induced apoptosis and cytotoxicity of chemotherapeutic agent bleomycin were then examined and compared. RESULTS: DDH was found highly expressed by SCC A431 cells, which was barely detectable in other normal or malignant cutaneous cells, including keratinocytes, fibroblast, and basal cell carcinoma cell line. RNAi Inhibition of DDH expression in A431 cells led to increased sensitivity to UVB-induced apoptosis and cytotoxicity of bleomycin treatment. CONCLUSION: DDH may play important roles in tumor progression of SCC via induction of apoptosis- and drug-resistance. PMID- 16361084 TI - Human keratinocyte radiosensitivity is linked to redox modulation. AB - BACKGROUND: Ionising radiation-induced reactive oxygen species (ROS) overproduction induces keratinocyte alterations and constitutes one of the most common effects after therapeutic gamma-irradiation. ROS production is controlled by a complex enzymatic system. OBJECTIVE: The aim of our study is to analyse the role of radiation-induced oxidative stress in keratinocytes death by apoptosis. We hypothesized that keratinocyte capacity to hamper radiation-induced ROS generation may control their radiosensitivity. METHODS: For this purpose, an original human skin explant model was developed and two types of human epidermal cells were used: primary keratinocytes NHEK and spontaneous non-tumourigenic cell line HaCaT. RESULTS: cDNA-arrays analysis was performed 24h after a 20Gy gamma radiation and revealed down-regulation of genes involved in oxidative stress control and the apoptosis process. This was confirmed by alterations in catalase, GPx and SOD enzymatic activities. This redox modulation was concomitant to the down-regulation of anti-apoptotic genes and up-regulation of some pro-apoptotic genes (caspase 10, ubiquitin C). Interestingly TUNEL labelling revealed an increase in the number of apoptotic cells. We also demonstrated a differential inducibility of the cell antioxidant network in two keratinocyte lines, which results in a differential cellular level of ROS, explaining their different radiosensitivities. CONCLUSION: Keratinocytes apoptosis is partly dependent on ROS production after exposure to gamma-rays. In addition, the differential radiosensitivity of keratinocytes is linked to different oxidative stress responses. PMID- 16361085 TI - PET neuroimaging of [11C]mirtazapine enantiomers in pigs. AB - Previously, we used positron emission tomography (PET) for studying the pharmacokinetics of rac-[11C]mirtazapine in living brain. Our findings showed that rac-[11C]mirtazapine has suitable properties for PET neuroimaging. However, separate studies of enantiomers are typically required for characterizing the pharmacokinetics of a racemic drug. Therefore, we have determined the whole-body distribution and brain pharmacokinetics of S- and R-[11C]mirtazapine in pigs. The enantiomers of [11C]mirtazapine produced similar effective doses of radioactivity in most body organs, except for the brain, in which the dose was approximately 40% higher after injection of S-[11C]mirtazapine than the antipode. Kinetic analyses of dynamic brain PET recordings showed that values for regional accumulation of compound (k3) were significantly higher for S-[11C]mirtazapine than for the antipode, while the values for clearance of compounds from tissue to circulation (k2) were consistently lower for S-[11C]mirtazapine than for the R form. No reliable difference occurred in the rate of metabolism of S- and R [11C]mirtazapine in the bloodstream of the pigs. The present findings indicate that enantioselective processes affect the cerebral pharmacokinetics of rac mirtazapine. PMID- 16361087 TI - Evidence of resilience: neuroimaging in former prisoners of war. AB - In this study, single voxel proton magnetic resonance spectroscopic imaging ((1)H MRS) and volumetric analysis of hippocampal magnetic resonance imaging (MRI) images were used to determine if any differences in hippocampal biochemistry or volume were present between former prisoners of war (POWs) with and without posttraumatic stress disorder (PTSD) and control subjects matched for age and education. This study did not find lower hippocampal concentrations of N acetylaspartate (NAA), smaller hippocampal volumes, or more impaired memory function in older veterans with PTSD compared with a group matched for traumatic experience or a nontraumatized control group. PMID- 16361086 TI - A systematic review of the effectiveness of naltrexone in the maintenance treatment of opioid and alcohol dependence. AB - This systematic review summarises evidence of the effectiveness of naltrexone (NTX) and the added value of psychosocial treatment in the maintenance treatment of opioid and alcohol dependence. Studies were selected through a literature search conducted in March 2004. Seven opioid and seventeen alcohol studies were identified. When possible, meta-(regression) analyses were performed. There is lack of evidence about the effectiveness of NTX in the maintenance treatment of opioid dependence. There is evidence for the effectiveness and applicability of NTX in the management of alcohol dependence. The opioid studies combined NTX with a variety of psychosocial interventions, which plagued the evaluation of their value. Concomitant psychosocial interventions used in the alcohol studies were mainly cognitive behavioural, which seems to be more effective than NTX combined with supportive therapy. Available data do not allow firm conclusions regarding the added effect of psychosocial interventions. However, the data suggest that a combination of naltrexone with cognitive behavioural relapse prevention therapy is beneficial in alcohol dependent patients. PMID- 16361088 TI - Striatal dopamine D1 and D2 receptor balance in twins at increased genetic risk for schizophrenia. AB - The dopamine hypothesis of schizophrenia postulates that a dysfunctional dopaminergic system is a major pathophysiological mechanism in the disease. Most studies have focused on striatal dopamine D2 receptors, but a disturbed link between dopamine D1 and D2 receptors has also been proposed. Schizophrenia is highly heritable, and recent evidence suggests that alterations in the dopaminergic system confer susceptibility for schizophrenia instead of being solely related to the to overt expression of the disease. To explore the impact of genetic vulnerability for schizophrenia on the balance of striatal dopamine D1 and D2 receptors, we studied monozygotic (MZ) and dizygotic (DZ) unaffected co twins from twin pairs discordant for schizophrenia as well as healthy control twins using positron emission tomography (PET). Both [(11)C]SCH 23390 and [(11)C]raclopride were used to quantitate D1 and D2 receptor binding, respectively, in the same individuals during the same day. The association between D1 and D2 receptor binding was analyzed using conventional region of interests as well as voxel-wise D1/D2 ratio maps. All levels of analyses failed to show any differences in D1/D2 ratio between the unaffected MZ or DZ co-twins and control twins. We noted rostrocaudally declining and dorsoventrally increasing gradients in D1/D2 ratio in the striatum, with no differences between groups in these gradients. In this sample, we did not find evidence for an association between increased genetic risk for schizophrenia and altered D1/D2 receptor balance in the striatum. PMID- 16361089 TI - Prefrontal subregions and dimensions of insight in first-episode schizophrenia--a pilot study. AB - Deficits in insight are multidimensional, and include symptom unawareness and misattribution. We and others have observed that these deficits may be related to a prefrontal dysfunction. However, few studies have examined the relationship between specific prefrontal sub-regions and the awareness and attributional dimensions of insight in schizophrenia. This study examined the correlation between insight dimensions of awareness and attribution of symptoms and dorsolateral prefrontal cortex (DLPFC) and orbitofrontal cortex (OFC) volume in 14 subjects with first-episode, antipsychotic-naive (FEAN) schizophrenia. In addition, 21 healthy subjects provided control data for volumetric assessments. Insight was assessed with Scale to Assess Unawareness of Mental Disorders. Morphometric assessments were adjusted for intra-cranial volume and were conducted by trained raters blind to clinical information using BRAINS-2. Average scores on current awareness of symptoms (1=aware; 5=unaware) were negatively correlated with right DLPFC volume and average scores on current attribution of symptoms (1=attribute; 5=misattribute) with right medial OFC volume. Unawareness and misattribution of symptoms in FEAN schizophrenia may have distinct neuroanatomical bases. DLPFC deficits may have resulted in illness unawareness by interfering with self-monitoring, while OFC abnormalities may have mediated symptom misattribution by conferring aberrant salience to perceived symptomatology. PMID- 16361090 TI - Spectral spin diffusion and magnetic dipolar energy in the NMR of 13CH3 compounds. AB - Spin diffusion between 13CH3 groups in solids is studied both theoretically and experimentally. It is shown to be dominated by mutual spin flip-flops of protons belonging to neighbouring methyl groups. Also nonmethyl protons may contribute significantly if present in the sample. The spin-rotational ground state of 13CH3 consists of 16 sublevels. When their populations are used to describe spin diffusion, eight population combinations are shown to be important, two of them corresponding to the 13C-proton and proton-proton intra-methyl magnetic dipolar energies, Dc and Dp, respectively. Spin-diffusion transitions modulate these combinations so that a further reduction to two sets of four combinations is possible, with no coupling between the sets. Coupled differential equations are derived to describe the time dependence of the combinations in each set. They are solved numerically and compared with experimental results on a single crystal of aspirin with 13C-labelled methyl groups at the carbon resonance. The 13C NMR induction signal was observed as a function of time after the preparation either at the carbon resonance (a two-pulse sequence) or at the proton resonance (proton saturation). Usually carbon spectra were computed first and then three of the mentioned population combinations were obtained from the individual spectral components. Some results on the time dependence of Dc were also obtained directly from the amplitude of the out-of-phase induction signal. Theoretical predictions are found to describe semiquantitatively the overall time dependence of these three combinations and especially their variation with different initial conditions, which are discussed in detail. Also the partial transfer of the magnetic dipolar energy between Dc and Dp is nicely explained. Reasons for discrepancies are discussed. PMID- 16361091 TI - Controlling centrosome number: licenses and blocks. AB - Centrosomes organize microtubule structures in animal cells. The centrosome duplicates once per cell cycle in most dividing cells via a pathway that relies on a pre-existing centrosome. The molecular mechanism of this 'once and only once' control is not understood, and recent results show that centrosomes can also be assembled by a de novo pathway that bypasses this control. These results require a rethinking of how proper centrosome number is maintained. We propose that the engagement of centrioles with each other normally blocks centrosome re duplication, and that disengagement of centrioles from each other at the end of mitosis licenses them for duplication in the subsequent cell cycle. PMID- 16361092 TI - Kinesin's moonwalk. AB - Kinesin-1 is a single-molecule walking machine, driven by ATP turnover. Recent optical trapping experiments show that pulling backwards on a walking kinesin-1 molecule causes the mechanical walking action to reverse, while the coupled chemical cycle of ATP turnover continues, apparently, to run forwards -- kinesin can moonwalk. Individual forward- and back-steps are fast, and each appears to be a single event, complete in a few tens of microseconds, with no substeps. Between steps, kinesin pauses, waiting for the next ATP to arrive. Several lines of evidence indicate that during these between-step dwells, only one of the two heads is strongly attached to the microtubule. The position of the other head during the dwells is less certain, and more controversial. PMID- 16361093 TI - Asymmetric spindle positioning. AB - When a spindle is positioned asymmetrically in a dividing cell, the resulting daughter cells are unequal in size. Asymmetric spindle positioning is driven by regulated forces that can pull or push a spindle. The physical and molecular mechanisms that can position spindles asymmetrically have been studied in several systems, and some themes have begun to emerge from recent research. Recent work in budding yeast has presented a model for how cytoskeletal motors and cortical capture molecules can function in orienting and positioning a spindle. The temporal regulation of microtubule-based pulling forces that move a spindle has been examined in one animal system. Although the spindle positioning force generators have not been identified in most animal systems, the forces have been found to be regulated by both PAR polarity proteins and G-protein signaling pathways in more than one animal system. PMID- 16361094 TI - MicroRNAs as oncogenes. AB - MicroRNAs (miRNAs) are a class of non-coding RNAs that function as endogenous triggers of the RNA interference pathway. Originally discovered in Caenorhabditis elegans, this group of tiny RNAs has moved to the forefront of biology. With over 300 miRNA genes identified in the human genome, and a plethora of predicted mRNA targets, it is believed that these small RNAs have a central role in diverse cellular and developmental processes. Concordant with this, aberrant expression of miRNA genes could lead to human disease, including cancer. Although the connection of miRNAs with cancer has been suspected for several years, four recent studies have confirmed the suspicion that miRNAs regulate cell proliferation and apoptosis, and play a role in cancer. PMID- 16361095 TI - Molecular mechanisms of dendritic spine morphogenesis. AB - Excitatory synapses are formed on dendritic spines, postsynaptic structures that change during development and in response to synaptic activity. Once mature, however, spines can remain stable for many months. The molecular mechanisms that control the formation and elimination, motility and stability, and size and shape of dendritic spines are being revealed. Multiple signaling pathways, particularly those involving Rho and Ras family small GTPases, converge on the actin cytoskeleton to regulate spine morphology and dynamics bidirectionally. Numerous cell surface receptors, scaffold proteins and actin binding proteins are concentrated in spines and engaged in spine morphogenesis. PMID- 16361096 TI - Phosphate removal by refined aspen wood fiber treated with carboxymethyl cellulose and ferrous chloride. AB - Biomass-based filtration media are of interest as an economical means to remove pollutants and nutrients found in stormwater runoff. Refined aspen wood fiber samples treated with iron salt solutions demonstrated limited capacities to remove (ortho)phosphate from test solutions. To provide additional sites for iron complex formation, and thereby impart a greater capacity for phosphate removal, a fiber pretreatment with an aqueous solution of a non-toxic anionic polymer, carboxymethyl cellulose (CMC), was evaluated. Problems with excessive viscosities during the screening of commercially available CMC products led to the selection of an ultra low viscosity CMC product that was still usable at a 4% concentration in water. Soxhlet extractions of chipped aspen wood and refined aspen wood fiber samples showed a higher extractives content for the refined material. Analysis of these extracts by FTIR spectroscopy suggested that the higher extractives content for the refined material resulted from the fragmentation of cell wall polymers (e.g., lignin, hemicelluloses) normally insoluble in their native states. Spectroscopic analysis of CMC and ferrous chloride treated fibers showed that the complex formed was sufficiently stable to resist removal during subsequent water washes. Equilibrium sorption data, which fit better with a Freundlich isotherm model than a Langmuir isotherm model, showed that phosphate removal could be enhanced by the CMC pretreatment. Results suggest that the process outlined may provide a facile means to improve the phosphate removal capacity of biomass-based stormwater filtration media. PMID- 16361097 TI - Synthesis and DNA-binding ability of pyrrolo[2,1-c][1,4]benzodiazepine-azepane conjugates. AB - A series of pyrrolobenzodiazepine-azepane conjugates linked through different alkane spacers have been prepared and their DNA thermal denaturation studies have been carried out. One of the compound (4b), elevates the DNA helix melting temperature of the CT-DNA by 2.0 degrees C after incubation for 36 h at 37 degrees C. PMID- 16361098 TI - Synthesis and biological evaluation of novel hexahydro pyrido[3',2':4,5]pyrrolo[1,2-a]pyrazines as potent and selective 5-HT(2C) receptor agonists. AB - Further lead optimization efforts on previously described 1,2,3,4,10,10a hexahydro-1H-pyrazino[1,2-a]indoles led to the new class of 5,5a,6,7,8,9 hexahydro-pyrido[3',2':4,5]pyrrolo[1,2-a]pyrazines culminating in the discovery of (5aR,9R)-2-[(cyclopropylmethoxy)methyl]-5,5a,6,7,8,9-hexahydro-9-methyl pyrido[3', 2':4,5]pyrrolo[1,2-a]pyrazine 18 as a potent, full 5-HT(2C) receptor agonist with an outstanding selectivity profile and excellent hERG and phospholipidosis properties. PMID- 16361099 TI - Probing structure/affinity relationships for the Plasmodium falciparum hexose transporter with glucose derivatives. AB - A series of 3-O-substituted glucose derivatives was prepared with alkyl, alkenyl, aromatic and ferrocenic substituents; to vary lipophilicity and hydrogen bonding ethylenedioxy and perfluorinated fragments were also introduced. Apparent affinities for the Plasmodium falciparum hexose transporter (PfHT) were determined after heterologous expression in Xenopus oocytes, with highest affinities for compounds with C8-C13 lipophilic chains. As no derivatives show significant affinity for the mammalian glucose transporter (GLUT1), these structure/affinity assays contribute to design of potent PfHT inhibitors and eventual development of antimalarials. PMID- 16361100 TI - Structure-activity studies of quinuclidinone analogs as anti-proliferative agents in lung cancer cell lines. AB - We have synthesized and tested novel quinuclidinone analogs to assay the effects on H1299 lung cancer cell lines alone or with gamma-radiation. We have found two series of quinuclidinone analogs that act as anti-cancer agents. Of these, four interesting analogs significantly decreased cell viability in H1299 lung cancer cell lines. Two derivatives decreased cell proliferation in a dose-dependent fashion alone or in the presence of gamma-radiation. Radiosensitization increased when derivative treatment preceded radiation treatment for both derivatives. These preliminary studies show an evidence for both additive and synergistic cytotoxicity for treatment of lung cancer by these novel quinuclidinone analogs. PMID- 16361101 TI - Immediate structural changes of porcine renal arteries after angioplasty: a histological and morphometric study. AB - The aim of this research was to characterize the immediate alterations induced by angioplasty and to compare the results of the application of two types of balloons. Ten porcine renal arteries were dilated with a compliant balloon, and ten with a non-compliant balloon. After angioplastic treatment arterial specimens were wax embedded for light microscopy. Sections were stained with the orcein-Van Gieson method, orcein, haematoxylin-eosin, and PAS. Image analysis was performed taking into consideration the following parameters: thickness of the entire wall, of the tunica media and of the inner elastic lamina. The major axes of the smooth muscle cells nuclei were also measured. The effects of the two types of balloon resulted in changes consisting in thinning of the entire arterial wall, reduction of the tunica media, distension of reticular fibers, presence of wide spaces between smooth muscle cells, stretching of smooth muscle cells, inner elastic lamina thickening. Both angioplasty devices used can modify the vascular wall. The identification of the tunica media structural damages might be useful in order to estimate the behavior of the vascular wall in the follow-up after angioplasty, because the entity of modifications could be predictive of restenosis that often takes place weeks or months after angioplasty. PMID- 16361102 TI - Histology and histochemistry of the ventriculus of Dolichoderus (=Monacis) bispinosus (OLIVIER, 1792) (Hymenoptera: Formicidae). AB - In this study we histologically and histochemically describe the ventriculus of Dolichoderus bispinosus. The epithelium consists of two basic cell types, highly basophilic generative cells, and digestive cells. The latter present several cytoplasmic vesicles, rich in acidic and neutral polysaccharides, and basic proteins. Also, these cells exhibit an apocrine secretion pattern. A mass of fibrous material is observed on the surface of the epithelium. Finally, we discuss the results obtained. PMID- 16361103 TI - Application of transmission electron microscopy to the clinical study of viral and bacterial infections: present and future. AB - Transmission electron microscopy has had a profound impact on our knowledge and understanding of viruses and bacteria. The 1000-fold improvement in resolution provided by electron microscopy (EM) has allowed visualization of viruses, the existence of which had previously only been suspected as the causative agents of transmissible infectious disease. Viruses are grouped into families based on their morphology. Viruses from different families look different and these morphological variances are the basis for identification of viruses by EM. Electron microscopy initially came to prominence in diagnostic microbiology in the late 1960s when it was used in the rapid diagnosis of smallpox, by differentiating, on a morphological basis, poxviruses from the less problematic herpesviruses in skin lesions. Subsequently, the technique was employed in the diagnosis of other viral infections, such as hepatitis B and parvovirus B19. Electron microscopy has led to the discovery of many new viruses, most notably the various viruses associated with gastroenteritis, for which it remained the principal diagnostic method until fairly recent times. Development of molecular techniques, which offer greater sensitivity and often the capacity to easily process large numbers of samples, has replaced EM in many areas of diagnostic virology. Hence the role of EM in clinical virology is evolving with less emphasis on diagnosis and more on research, although this is likely only to be undertaken in specialist centres. However, EM still offers tremendous advantages to the microbiologist, both in the speed of diagnosis and the potential for detecting, by a single test, any viral pathogen or even multiple pathogens present within a sample. There is continuing use of EM for the investigation of new and emerging agents, such as SARS and human monkeypox virus. Furthermore, EM forms a vital part of the national emergency response programme of many countries and will provide a frontline diagnostic service in the event of a bioterrorism incident, particularly in the scenario of a deliberate release of smallpox virus. In the field of bacteriology, EM is of little use diagnostically, although some bacterial pathogens can be identified in biopsy material processed for EM examination. Electron microscopy has been used, however, to elucidate the structure and function of many bacterial features, such as flagellae, fimbriae and spores and in the study of bacteriophages. The combined use of EM and gold labelled antibodies provides a powerful tool for the ultrastructural localisation of bacterial and viral antigens. PMID- 16361104 TI - High resolution morphological analysis of in situ human chromosomes. AB - The purpose of this study was to analyze the inner structure of chromosomes in cells arrested, fixed and cryosectioned in metaphase. The chromosomes in metaphase maps prepared using standard cytogenetic protocols, are usually covered by cellular debris, which obscures the structural details on the surface and limits analysis by techniques when using nanometric resolution. By using cryosectioning, the debris is removed and it is possible to analyze the internal structure of the chromosomes. We described the ultrastructure of chromosome sections fixed with either acetic acid, methanol or glutaraldehyde, evaluating the effect and the influence of the fixative on the morphology. Furthermore, we subjected those cells previously fixed with glutaraldehyde to osmic maceration in order to better visualize the intracellular structure. All samples were examined with a Field Emission In Lens Scanning Electron Microscope (FEISEM), which allows high-resolution analysis of biological samples without any metal coating. The results showed a package morphology in samples fixed with glutaraldehyde, mainly due to the high capacity of the fixative to strongly crosslink the proteins. In contrast, the fibrillar structure seen in cryosections fixed with acetic acid/methanol is due to the propensity of the fixatives to extract and remove proteins. We propose that in situ chromosomes fixed with glutaraldehyde and then osmicated are a good model for studying the inner structure of chromosomes by using high resolution scanning electron microscopy. PMID- 16361105 TI - Improved separation methods for the recovery of (82)Sr from irradiated targets. AB - (82)Rb is a short-lived positron-emitting isotope (T(1/2)=75 s) that is increasingly being used in PET to study blood flow through the heart and brain. This isotope is supplied to physicians in the form of a generator where the parent isotope, (82)Sr, is immobilized onto an ion exchange column allowing the (82)Rb to be eluted with isotonic saline as required. (82)Sr is manufactured by the proton irradiation of molybdenum, rubidium chloride or rubidium metal targets followed by complex separations to recover the (82)Sr. A number of inorganic ion exchange materials were evaluated for their ability to remove (82)Sr from simulated target solutions and compared with currently used organic ion exchange resins. Sodium nonatitanate was identified as a replacement for current resins as a potential material that would simplify target processing and improve the yield of (82)Sr from each target. PMID- 16361106 TI - PTD-XIAP protects against cerebral ischemia by anti-apoptotic and transcriptional regulatory mechanisms. AB - Caspases play a major role in the infarction process that follows occlusion of cerebral arteries and are important targets for stroke therapy. We have generated three fusion proteins that link various domains of the X chromosome-linked inhibitor of apoptosis (XIAP), a potent caspase inhibitor, to the protein transduction domain (PTD) of HIV-1/Tat, and have tested their efficacy after distal occlusion of the middle cerebral artery (dMCAO) in mice. PTD-XIAP failed to accumulate in brain structures after intravenous (iv) delivery, but properly transduced cortical cells when applied topically. Shorter constructs efficiently targeted the lesion after iv delivery. All proteins retained their caspase inhibitory activity and significantly reduced infarct volumes. PTD-XIAP reversed long-term impairments in the water maze test. Sequential activation of transcription factors was observed, suggesting that the effects of XIAP are mediated by both direct inhibition of apoptotic mechanisms and secondary regulation of transcription factors involved in neuronal survival. PMID- 16361107 TI - Dystonia-causing mutant torsinA inhibits cell adhesion and neurite extension through interference with cytoskeletal dynamics. AB - Early onset torsion dystonia is a movement disorder inherited as an autosomal dominant syndrome with reduced penetrance. Symptoms appear to result from altered neuronal circuitry within the brain with no evidence of neuronal loss. Most cases are caused by loss of a glutamic acid residue in the AAA+ chaperone protein, torsinA, encoded in the DYT1 gene. In this study, torsinA was found to move in conjunction with vimentin in three cell culture paradigms-recovery from microtubule depolymerization, expression of a dominant-negative form of kinesin light chain and respreading after trypsinization. Co-immune precipitation studies revealed association between vimentin and torsinA in a complex including other cytoskeletal elements, actin and tubulin, as well as two proteins previously shown to interact with torsinA-the motor protein, kinesin light chain 1, and the nuclear envelope protein, LAP1. Morphologic and functional differences related to vimentin were noted in primary fibroblasts from patients carrying this DYT1 mutation as compared with controls, including an increased perinuclear concentration of vimentin and a delayed rate of adhesion to the substratum. Overexpression of mutant torsinA inhibited neurite extension in human neuroblastoma cells, with torsinA and vimentin immunoreactivity enriched in the perinuclear region and in cytoplasmic inclusions. Collectively, these studies suggest that mutant torsinA interferes with cytoskeletal events involving vimentin, possibly by restricting movement of these particles/filaments, and hence may affect development of neuronal pathways in the brain. PMID- 16361108 TI - Enriched environment and spatial learning enhance hippocampal neurogenesis and salvages ischemic penumbra after focal cerebral ischemia. AB - Enriched environment (EE) has been shown to increase neurogenesis in the adult brain. The aim of this study is to determine the effect of EE and spatial learning on neurogenesis following ischemic stroke. Male adult SD rats were subjected to sham surgery or distal middle cerebral artery occlusion (MCAO). MCAO induced a transient increase followed by a sustained depression of progenitor cell proliferation and neuroblast production below baseline level in both ipsilateral and contralateral DG compared to sham. Increased neuronal differentiation and neurogenesis in the DG were observed in both sham and MCAO rats following 8 weeks in the EE combined with spatial learning, compared to rats housed in the standard environment. EE/Learning also restored the total number of neuroblasts in the DG after MCAO compared to sham. Furthermore, EE/learning enhanced the density of NeuN positive cells in the ischemic penumbra, though no new neurons were detected in this region. PMID- 16361109 TI - Structural analysis of the ZEN-4/CeMKLP1 motor domain and its interaction with microtubules. AB - The centralspindlin complex is required for the assembly and maintenance of the central spindle during late anaphase and the completion of cytokinesis. It is composed of two copies each of the kinesin-like protein ZEN-4, a Caenorhabditis elegans MKLP-1 (Kinesin-6 family), and the RhoGAP CYK-4. By using cryo-electron microscopy and helical 3D reconstruction, we are investigating the structural features of the interactions between monomeric and dimeric motor domain constructs of ZEN-4 and microtubules. We have calculated helically averaged 3D maps of microtubules decorated with ZEN-4 motor domain in the presence of AMP PNP, ADP, ADP-AlF(4)(-), and nucleotide-free conditions. We used statistical difference mapping to compare these maps among each other and to related maps obtained from microtubules decorated with a well-characterized Kinesin-1 motor domain from Neurospora crassa. Thereby, we found distinct structural features in microtubule-ZEN-4 complexes that may directly relate to the functional properties of ZEN-4 and centralspindlin. Furthermore, we investigated the location, structure, and function of a highly conserved extension of approximately 50 residues unique to the Kinesin-6 subfamily, located in the motor core loop6/beta4 region. PMID- 16361110 TI - Changes in lipid and fatty acid composition of late larval and puerulus stages of the spiny lobster (Panulirus cygnus) across the continental shelf of Western Australia. AB - The feeding of the late larval stages of the spiny lobster, Panulirus cygnus, and the energy reserves used by the non-feeding post-larvae for crossing the continental shelf of Western Australia were examined through their lipid composition. Lipid was a significant component of the biomass of all larval and post-larval samples (range 63-213 mg g(-1) of dry biomass). The fatty acid profiles of late stage larvae (8-9) suggest that they were feeding on salps and small crustaceans, such as euphausids, from oligotrophic pelagic communities where herbivorous and microbial grazing is an important basis to the food web. There was a marked decrease in lipid content of post-larvae progressively across the continental shelf, and this corresponded closely with decreasing dry mass, suggesting that post-larvae were using lipid as an energy source during cross shelf movement at a rate of 1.6 J km(-1). This is considerably lower than for other spiny lobster species, suggesting that the post-larvae of the western rock lobster may use physical processes as well as active swimming for onshore transport. This may help to explain the large inter-annual variability in the post-larval settlement of this species, which is closely related to differences in weather patterns capable of greatly varying onshore advection processes. PMID- 16361111 TI - Transmission ecology of Echinococcus multilocularis: what are the ranges of parasite stability among various host communities in China? AB - A striking feature of the transmission ecology of Echinococcus multilocularis in China is the diversity of hosts that contribute to the parasite cycle. Considering the population dynamics of key reservoir intermediate hosts and the ratio of their preferred habitat in a landscape (ROMPA) is essential to understanding transmission, but the numerous communities in which the parasite cycles and the extent of those communities is currently far from being fully documented. On the Tibetan plateau grassland management influences intermediate host species populations and the sheer size of the area could be a major contributing factor to sustaining transmission in the region. In Southern Gansu and Southern Ningxia the processes of deforestation have temporarily provided optimal habitat for key reservoir intermediate hosts of E. multilocularis, resulting in high transmission and human disease, however currently the parasite may be extinct locally. Faced with this pattern of potentially transient transmission in a diversity of communities the question of the dispersal potential of the parasite arises. The reforestation program currently active across much of Western China has the potential to give rise to a massive increase in habitat favourable to suitable intermediate hosts and emergence or re emergence of the zoonosis alveolar echinococcosis in many areas. This potential epidemic could be mitigated by both natural and human induced parasite dispersal mechanisms including fox migration and the sale of infected dogs originating from stable endemic foci on the Tibetan plateau. However, currently the degree of genetic exchange between discrete transmission foci is unknown and it is expected that genetic techniques could provide crucial information regarding this important question. PMID- 16361112 TI - Present situation of cystic echinococcosis in Central Asia. AB - Cystic echinococcosis (CE) caused by Echinococcus granulosus has always been an endemic disease in central Asia. During the period of Soviet Administration up to 1991, human surgical incidence rates tended to be relatively low with perhaps at most 1-5 cases per 100,000 per year. Following the collapse of the Soviet Union and the emergence of the New Independent States there has been profound economic and social changes. Associated with this has been a serious epidemic of CE throughout the region. In many areas figures suggest the surgical incidence is now greater than 10 cases per 100,000. Furthermore, official government figures are believed to substantially under report the extent of the problem. For example, official figures in Uzbekistan reported 819 cases of CE surgically treated in 2001. However, a detailed analysis of hospital records suggests that the true figure was 4089, more than 4 fold higher. The latter figure represents an annual surgical incidence rate of nearly 25 cases/100,000 per year. Similarly high endemic areas are seen in southern Kazakhstan, Kyrgystan and Tadjikistan with incidence rates of up to 13 cases/100,000, 20 cases/100,000 and 27 cases/100,000 respectively. A disproportionate number of cases are in children and the unemployed. The rates of infection have also increased in major livestock species such as sheep with a doubling of reported prevalence in some areas. In the dog population, independent studies in Uzbekistan and Kazakhstan have demonstrated that the rural dog population, closely associated with the sheep industry, is highly infected, with prevalences approaching 25%. Village and urban dogs have a considerably lower prevalence. PMID- 16361113 TI - Epidemiology of Streptococcus agalactiae colonization in Germany. AB - Streptococcus agalactiae can cause severe pneumonia, sepsis and meningitis in neonates and remains one of the most prevalent causes of invasive neonatal infections. Maternal transmission of S. agalactiae during delivery can be prevented by prenatal screening and peripartal antibiotic prophylaxis. Implementation of CDC guidelines for group B streptococci (GBS) disease prevention resulted in a significant decline of invasive neonatal S. agalactiae infections in the USA. Similar national guidelines were issued in 2000 for Germany. However, the epidemiology of S. agalactiae colonization in Germany has not been investigated for more than 15 years and the impact these guidelines will have is therefore unknown. To assess colonization rates in Germany, we cultured vaginal and rectal swabs for S. agalactiae from pregnant and non-pregnant adult patients in the region of Aachen and Munich. Swabs were cultivated in selective broth medium for 24h and subsequently plated on blood agar plates according to the CDC recommendations. Colonies negative for catalase and pyrrolidonyl aminopeptidase were further differentiated by the CAMP test and a DNA probe specific for S. agalactiae. Rectal or vaginal colonization of S. agalactiae was found in 34 (16%) of 210 pregnant patients and in 41 (16%) of 250 non-pregnant women. S. agalactiae was found only in rectal swabs in 4% of pregnant and non pregnant patients. For further characterization of the strains capsular serotypes and major surface protein antigens were determined by Ouchterlony immunodiffusion and PCR. Among the 75 different patient isolates serotype III was the most prevalent with 21 (28%) isolates, followed by 16 (21%) isolates of serotype II, 13 (17%) isolates of serotype Ia, 12 (16%) of serotype V, 11 (15%) of serotype Ib and only 2 (3%) isolates of serotype IV. The vast majority of all strains harbored genes for the major surface protein antigens, the alpha-C-protein or alpha-C-protein like antigens like Alp2-4, epsilon and Rib. These data show that S. agalactiae colonization is common in Germany and strict adherence to the guidelines for the preventions of GBS disease will result in peripartal antibiotic prophylaxis in up to 20% of all deliveries. PMID- 16361114 TI - Kainate receptor physiology. AB - Glutamate receptors constitute a complex signalling system at most of the excitatory synapses in the brain. Of the known ionotropic glutamate receptors, kainate receptors are ubiquitous in the central nervous system, and a considerable amount of data indicates that this class of receptors is present at both sides of the synapse. Pre- and postsynaptic kainate receptors are able to regulate both transmission of information and excitability in a synapse-specific manner. Proteins interacting with kainate receptor subunits are being identified and functional studies have provided evidence of the existence of a dual signalling system. It has become clear that these receptors have a role in synaptic plasticity and that they might also have a fundamental role in epilepsy through the strategic control of network excitability. However, the role of kainate receptors in other brain pathologies remains obscure. PMID- 16361115 TI - GABAB receptor: a site of therapeutic benefit. AB - Although the presence of functional GABAB receptors in mammalian brain has been known for more than 20 years, there is still only one therapeutic agent in use, baclofen, which mediates its effects directly via this receptor. However, activation of this receptor can produce numerous effects that might be amenable to drug development. Evidence from preclinical studies also suggests that antagonism of the GABAB receptor produces beneficial clinical effects. PMID- 16361116 TI - Glutamate-based therapeutic approaches: ampakines. AB - Ampakines are a structurally diverse family of small molecules that positively modulate alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors, and thereby enhance fast, excitatory transmission throughout the brain. Surprisingly, ampakines have discrete effects on brain activity and behavior. Because their excitatory synaptic targets mediate communication between cortical regions, serve as sites of memory encoding, and regulate the production of growth factors, ampakines have a broad range of potential therapeutic applications. Several of these possibilities have been tested with positive results in preclinical models; preliminary clinical work has also been encouraging. PMID- 16361117 TI - Gene therapy strategies for Duchenne muscular dystrophy utilizing recombinant adeno-associated virus vectors. AB - Gene transfer vectors based on adeno-associated virus (AAV) are now widely used in the field of gene therapy. These vectors have been studied for their potential use in treating many diseases, among them the muscular dystrophies, the most common of which is Duchenne muscular dystrophy (DMD). Several recent advances in the areas of AAV serotype analysis, transgene engineering, and vector delivery to muscle, together with novel means of rescuing mutant mRNA transcripts, have yielded impressive results in animal models of DMD. This minireview focuses on these recent advances and their implications for potential treatments for DMD and other neuromuscular disorders. PMID- 16361118 TI - The role of echocardiography screening in athletes for cardiovascular disease. PMID- 16361119 TI - What is the relationship between gastro-oesophageal reflux and obstructive sleep apnoea? PMID- 16361120 TI - Transosseous application of low-intensity ultrasound for the enhancement and monitoring of fracture healing process in a sheep osteotomy model. AB - The purpose of this study is twofold: (a) to investigate the application of transosseous low-intensity pulsed ultrasound (LiUS) on the enhancement of fracture healing and (b) to demonstrate the ability of transosseous ultrasound propagation to monitor the healing process. A midshaft tibial osteotomy model was used on 40 skeletally mature sheep, and an external fixator was applied to maintain the reduction and stabilization of the osteotomy. Two ultrasound transducers were implanted into the fracture site in contact with the bone. For investigating the efficacy of LiUS, the animals were randomly divided in two equal groups: the treatment group and the control group. The LiUS-treated animals received 200-micros bursts of 1-MHz sine waves with a pulse repetition rate of 1 kHz and average intensity of 30 mW/cm2, for 20 min daily. For monitoring purposes, an ultrasound dataset was constructed consisting of serial ultrasound measurements obtained from healing bones. Animals' sacrifice took place on the 100th post-operative day. The effect of LiUS on fracture healing was evaluated using radiographs, destructive three-point bending testing and quantitative CT based bone mineral density (BMD) measurements. Survival analysis using Kaplan Meier curves showed significantly higher probability of radiographic healing for the animals in the treatment group (P = 0.009). Statistical significance was also observed for callus BMD (P = 0.003, Wilcoxon nonparametric test), the breaking load (P = 0.001), extrinsic stiffness (P = 0.019), Young's modulus (P = 0.043) and ultimate strength (P = 0.051) in favor of the LiUS-treated limbs. Analysis of the obtained ultrasonic measurements showed that the propagation velocity across healing bones constitutes a significant feature able to early distinguish between healed and nonhealed bones (area under ROC curve was 0.810 and 0.841 on the 80th and 100th post-operative days, respectively). Both the LiUS application and the ultrasonic measurements are supported by an integrated telemedicine system which also incorporates an ultrasound wearable device and a patient management system. PMID- 16361121 TI - Preservation of thoracic spine microarchitecture by alendronate: comparison of histology and microCT. AB - The effect of bisphosphonates on trabecular microarchitecture may contribute to the reduced risk of vertebral fracture with treatment independent of the bone volume. Trabecular structure was examined at the twelfth thoracic vertebra after 2 years of treatment of two groups of ovariectomized baboons on high and low doses of alendronate, compared with ovariectomized and non-ovariectomized controls. Standard 2D histological measurements showed that alendronate treatment of ovariectomized animals resulted in significantly higher total trabecular length and a lower marrow star volume in comparison with ovariectomized controls indicating preservation of connectivity. Similarly when the vertebrae were examined using a novel thick slice technique that combines 2D and 3D information, ovariectomy produced a significantly higher number of "real" trabecular termini in comparison with normal. When ovariectomized animals were treated with increasing doses of alendronate, fewer "real" termini were seen. MicroCT analysis (2D and 3D) correlated well with the histological measurements, although more variability and less discrimination between groups was seen, with no statistically significant differences with alendronate treatment. Reduced vertebral fracture risk with alendronate may be due to a combination of factors including the increased bone volume, reduced turnover and greater mineralization reported by others. Added to this is now suggested the preservation of several aspects of vertebral cancellous architecture, with microscopy the most sensitive method of analysis. PMID- 16361122 TI - What the clinician wants to know: radiation oncology perspective. AB - The delineation of the target volume, i.e. the volume which should be irradiated with a therapeutic dose of irradiation, is of utmost importance in radiotherapy. Modern imaging techniques cannot be missed in this process. Diagnosticians and radiation oncologists therefore should understand each other's needs and potential. PMID- 16361123 TI - Screening women at increased risk with MRI. AB - Breast cancer is the most common cancer affecting women. In the screening of women for breast cancer, mammography is the most used imaging modality. Women with an increased risk for getting breast cancer can develop a malignancy at a relatively young age compared to other women. The increased risk for developing breast cancer can usually be found in a positive familial history. This positive familial history is based on a gene mutation in 5-10% of cases. The most common gene mutations are BRCA1 and BRCA2. This risk makes it necessary to start screening these women at a young age. Mammography, however, has proven to be less reliable in younger women because its sensitivity is lowered due to the dense breast tissue often present in this group. MRI has a higher sensitivity for detecting breast cancer compared to mammography. MRI is not influenced by the density of the breast tissue. This makes breast MRI the best modality available for the screening of women with an increased risk for developing breast cancer. PMID- 16361124 TI - Quantitative CT analysis for assessing response in lymphoma (Cheson's criteria). AB - Standardized CT-based criteria used for lymphoma staging and follow-up and the current role of FDG-PET are reviewed. The current CT-based International Workshop Criteria (IWC) still have the main advantage of representing standardized criteria allowing comparability of clinical trials in patients with lymphoma. However, functional imaging with integrated IWC and FDG-PET provide more accurate response assessment, and challenge the current paradigm. Although integration of FDG-PET in IWC requires validation in a prospective trial with a large number of patients, new long-term clinical and therapeutic trials probably need to be designed using these new and hopefully standardized functional criteria. This potentially could allow a more risk-adapted approach to the treatment of aggressive lymphoma: intensive (reinforced) therapies for non-responders vs. less intensive therapies for good responders with the main goal of improved clinical outcome. PMID- 16361125 TI - MRI and PET in monitoring response in lymphoma. AB - The potential of FDG-PET and MRI in monitoring response to treatment in lymphoma is reviewed. Both FDG-PET and MRI can provide whole body imaging. Both also share the advantage of combining functional and anatomical information. At present, hybrid FDG-PET and MDCT is the best technique for monitoring response to treatment, especially early response to treatment. Early assessment of response to treatment has the potential to tailor therapy. MR imaging is useful especially in assessing bone marrow and central nervous system involvement. PMID- 16361126 TI - Role of imaging to choose treatment. AB - Radiologists perform various examinations at every step of lymphomas. The role of imaging is atypical or a 'classical' oncologic radiologist, as multiple non radiological criteria are combined to decide on treatments. A good knowledge of the practical use of the results helps the radiologist to seek the useful pieces of information. In treatment evaluation, uncertain complete response is only used in lymphomas. Imaging is changing, with the emergence of PET and whole body MRI but CT remains the key examination today. The WHO criteria are the only ones used to evaluate treatment results on CT, even though the use of PET is increasingly used, with better and better results. PMID- 16361128 TI - Colorectal cancer: what the clinician wants to know. AB - Colorectal cancer is a common and lethal disease. The adenoma-carcinoma sequence offers a window of opportunity in which the precursor lesion or early carcinoma can be removed endoscopically to prevent systemic disease. New and advanced techniques to improve endoscopic detection of precursor lesions are being developed. Other, less invasive screening methods are currently being developed and may become of use for population-based screening in the near future. Recently, important developments in the treatment (both surgical and chemotherapeutic) of colorectal cancer have occurred. The extent of the disease (stage) forms the basis for therapeutic decisions and accurate imaging is crucial. PMID- 16361127 TI - Problems monitoring response in multiple myeloma. AB - There has been a revolution in the treatment of multiple myeloma over the past decade. This article seeks to correlate advances in imaging with advances in treatment and to highlight how proper understanding of both is necessary for optimum management. PMID- 16361129 TI - Virtual colonoscopy for colorectal cancer screening: current status. AB - Computed tomography colonography (CTC) (also known as 'virtual colonoscopy') is a noninvasive method of imaging the colon using helical CT. Although CTC has been shown to be useful for certain clinical indications, it has not yet been endorsed as a colorectal cancer screening test. The purpose of this article is to review the current status of CTC for colorectal cancer screening. CTC is an accurate method to detect colonic polyps and to select patients who would benefit from colonoscopy. The major advantages of CTC over conventional colonography include its relatively low risk and greater tolerance by patients. In this article, the CTC procedure and results of clinical trials are reviewed, as well as potential pitfalls related to CTC performance and interpretation. Finally, radiation dose, the discovery of incidental extracolonic findings with CTC, bowel preparation methods, and computer-aided diagnosis are addressed. PMID- 16361130 TI - Colorectal cancer: the role of PET/CT in recurrence. AB - Many imaging modalities and scanning techniques, such as contrast enhanced CT, MRI and FDG-PET, are available for assessment of recurrent colorectal carcinoma. In addition, integrated PET/CT is becoming increasingly available. Intuitively, a synergistic combination of scanning characteristics sounds promising. However, the exact clinical value has not yet been fully established. The role of PET/CT image fusion must be weighed carefully against other available modalities. In this review we evaluate the potential of combined PET/CT in recurrent colorectal carcinoma. When available, PET/CT currently appears the diagnostic tool of choice. In the near future, combined PET/MRI may further enhance the diagnostic algorithm. PMID- 16361131 TI - Liver metastases of colorectal cancer: US, CT or MR? AB - In contrast to other extrahepatic malignancies many colorectal cancers can be cured even when there is metastatic spread to the liver. The diagnosis of liver metastases relies totally on imaging to decide which patients may be surgical candidates. The diagnostic value of ultrasound with contrast agents, multidetector CT and MR imaging with non-specific gadolinium chelates and liver specific contrast agent is discussed. Nowadays MDCT is the mainstay of staging and follow-up of these patients, because it provides good coverage of the liver and the complete abdomen and the chest in one session. MR imaging has been shown to be superior to helical CT in the preoperative assessment of colorectal liver metastases. Large studies are needed to define the role of MDCT vs. MRI staging in patients referred for resection of liver metastases. PMID- 16361132 TI - Translational molecular imaging for cancer. AB - Although most clinical diagnostic imaging studies employ anatomic techniques such as computed tomography (CT) and magnetic resonance (MR) imaging, much of radiology research currently focuses on adapting these conventional methods to physiologic imaging as well as on introducing new techniques and probes for studying processes at the cellular and molecular levels in vivo, i.e. molecular imaging. Molecular imaging promises to provide new methods for the early detection of cancer and support for personalized cancer therapy. Although molecular imaging has been practiced in various incarnations for over 20 years in the context of nuclear medicine, other imaging modalities have only recently been applied to the noninvasive assessment of physiology and molecular events. Nevertheless, there has been sufficient experience with specifically targeted contrast agents and high-resolution techniques for MR imaging and other modalities that we must begin moving these new technologies from the laboratory to the clinic. This brief review outlines several of the more promising areas of pursuit in molecular imaging for oncology with an emphasis on those that show the most immediate likelihood for clinical translation. PMID- 16361133 TI - Molecular imaging: what can be used today. AB - Biochemical cellular targets and more general metabolic processes in cancer cells can be visualised. Extensive data are available on molecular imaging in preclinical models. However, innovative tracers move slowly to the clinic. This review provides information on the currently available methods of metabolic imaging, especially using PET in humans. The uptake mechanisms of tracer methods and a brief discussion of the more 'molecular' targeted methods are presented. The main focus is on the different classes of tracers and their application in various types of cancer within each class of tracers, based on the current literature and our own experience. Studies with [18F]FDG (energy metabolism), radiolabelled amino acids (protein metabolism), [18F]FLT (DNA metabolism), [11C]choline (cell membrane metabolism) as general metabolic tracer methods and [18F]DOPA (biogenic amine metabolism) as a more specific tracer method are discussed. As an example, molecular imaging methods that target the HER2 receptor and somatostatin receptor are described. PMID- 16361134 TI - The sentinel node in breast cancer: an update. AB - There has been rapid acceptance of sentinel lymph node biopsy into the management of breast cancer over the past 10 years. This article seeks to highlight the controversies and to summarise its current status. PMID- 16361136 TI - Oral cavity cancer. AB - Imaging plays a crucial role in the staging of oral cancers. Imaging information is essential for determining tumour resectibility, post resection surgical reconstruction and radiation therapy planning. The aim of this paper is to highlight the natural history of oral cancer spread and how malignant infiltration can be accurately mapped. It focuses on buccal mucosa, hard palate, tongue and floor of mouth carcinoma. PMID- 16361135 TI - What the clinician wants to know: surgical perspective and ultrasound for lymph node imaging of the neck. AB - Imaging of lymph node metastases in the neck can have two major indications: (1) prognosis and assisting with choice of treatment; (2) staging and detection of clinically occult metastases in different levels of the neck. Both indications are discussed. The role and limitations of US and US-guided fine-needle aspiration cytology are also reviewed. PMID- 16361137 TI - Oropharyngeal cancer. AB - Imaging studies have an important role in defining the extent of oropharyngeal neoplasms and coming to an accurate staging of these lesions. Besides influencing treatment choice, imaging studies can also be used to monitor tumour response to treatment, and as an adjunct to clinical follow-up in order to detect treatment failure as early as possible. PMID- 16361138 TI - Lymph node imaging: multidetector CT (MDCT). AB - Advances in cross-sectional imaging, including conventional and helical (spiral) CT and multidetector (MDCT) and MR imaging, now allow detailed evaluation of the anatomy and pathology of the neck and thoracic inlet. The major structures are identified by their appearance and that of contrasting fatty tissue planes surrounding the soft tissues. These structures include the larynx, trachea, thyroid, and parathyroid glands as well as the vessels, lymph node chains, nerves, and supporting muscles. A thorough understanding of the normal cross sectional anatomy is fundamental to properly interpret pathologic processes. Pathologic processes include both solid and cystic masses. Most solid masses are enlarged lymph nodes. In contrast, cystic masses are of variable pathology, and their characteristic appearances and locations with respect to normal neck anatomy allow a confident diagnosis to be made from a brief differential diagnostic spectrum. PMID- 16361139 TI - The clinical role of CT/PET in oncology: an update. AB - FDG-PET imaging has significantly altered the workup of the oncologic patient. With the introduction of combined FDG-PET/CT scanners, there has been a dramatic improvement in lesion detection, lesion characterization and accurate lesion localization. As attenuation correction can be attained with the CT images, PET exam times have been dramatically reduced, by as much as 50% in many instances. In this overview we briefly outline the advantages of CT/PET in oncology especially in diagnosis and clinical management of the common tumor types for which it is mainly used. PMID- 16361140 TI - Prostate ultrasound--for urologists only? AB - The value of ultrasound (US) in the diagnosis of prostate cancer has dramatically increased in the past decade. This is mainly related to the increasing incidence of prostate cancer, the most common cancer in men and one of the most important causes of death from cancer in men. The value of conventional gray-scale US for prostate cancer detection has been extensively investigated, and has shown a low sensitivity and specificity. Therefore conventional gray-scale US is mainly used by urologists for guiding systematic prostate biopsies. With the development of new US techniques, such as color and power Doppler US, and the introduction of US contrast agents, the role of US for prostate cancer detection has dramatically changed. Advances in US techniques were introduced to further increase the value of US contrast agents. Although most of these developments in US techniques, which use the interaction of the contrast agent with the transmitted US waves, are very sensitive for the detection of microbubbles, they are mostly unexplored, in particular for prostate applications. Early reports of contrast-enhanced US investigations of blood flow of the prostate have shown that contrast-enhanced US adds important information to the conventional gray-scale US technique. Furthermore, elastography or 'strain imaging' seems to have great potential in prostate cancer detection. Since these new advances in US are very sophisticated and need a long learning curve, radiologists, who are overall better trained with these new US techniques, will play a more important role in prostate cancer diagnosis. Current trends show that these new US techniques may allow for targeted biopsies and therefore replace the current 'gold standard' for prostate cancer detection--the systematic biopsy. Consequently the use of these new US techniques for the detection and clinical staging of prostate cancer is promising. However, future clinical trials will be needed to determine if the promise of these new US advances of the prostate evolves into clinical application. PMID- 16361141 TI - Doctor-patient relationship in oncologic radiology. AB - Progress in medicine and changes in our society have led to an increasing number of patients with cancer and a change in the doctor-patient relationship. Patients rights are now defined in several countries by laws. The course of cancer involves numerous imaging examinations in which the radiologist is primarily involved. It is often the radiologist who discovers abnormalities and who must break the news to the patient. This task is made all the more difficult by the radiologist's lack of specific training in the management of difficult situations such as announcing bad news. There is a high risk of inappropriate responses that can have a seriously damaging effect on the patient's state of mind. Even with the best intentions, it can be very profitable to review and improve our relational modalities and to more effectively meet the patient's increasing demand for information. The radiologist's technical know-how is not sufficient, as he must also be able to give just the right amount of information based on his clinical competence, and his relationship with patients while respecting their wishes and their rights. PMID- 16361142 TI - ACRIN--lessons learned in conducting multi-center trials of imaging and cancer. AB - The American College of Radiology Imaging Network (ACRIN) is a US National Cancer Institute-funded clinical trials cooperative group charged with conducting multi center clinical trials of diagnostic imaging and image-guided treatment technologies as they are employed in the detection, diagnosis and staging, treatment, and evaluation of treatment for cancer. Operating since 1999, ACRIN involves participating institutions around the world and hundreds of radiologists, methodologists, and scientists in the 22 trials it has been working on to date, including several large screening trials. The experience with ACRIN has elucidated the unique requirements that must be fulfilled by imaging trials if they are to be successful, particularly in such areas as trials design, definition of technologies and their findings, quality assurance, and ensuring sufficient accrual. ACRIN is now pursuing several courses of action that will disseminate the products devolving from ACRIN trials into the public domain, so that they may benefit other investigators. PMID- 16361143 TI - Imaging of thymic disorders. AB - Evaluation of the thymus poses a challenge to the radiologist. In addition to age related changes in thymic size, shape, and tissue composition, there is considerable variability in the normal adult thymic appearance within any age group. Many different types of disorders may affect the thymus, including hyperplasia, cysts, and benign and malignant neoplasms, both primary and secondary; clinical and imaging findings typical for each disease process are described in this article. Whereas computed tomography is the mainstay for imaging the thymus, other imaging modalities may occasionally provide additional structural or functional information. PMID- 16361144 TI - The radiology of gastrointestinal stromal tumours (GIST). AB - Gastrointestinal stromal tumours (GISTs) comprise a group of smooth muscle mesenchymal alimentary tract tumours of variable malignancy. Recently, the pathophysiology and radiology of these tumours has generated enormous interest following the discovery of a specific, highly effective, chemotherapeutic agent in the form of ST-571 (Imatinib; Glivec, Novartis, Frimley UK). At the time of this review, 106 patients with malignant gastrointestinal stromal tumours seen at the Royal Marsden Hospital have been entered into trials examining the efficacy of varying doses of Imatinib. Burkill et al., also from the Royal Marsden Hospital, have previously reported the distribution, imaging features and pattern of metastatic spread of these tumours (Burkill GJ, Badran M, Al-Muderis O et al. Malignant gastrointestinal stromal tumor: distribution, imaging features, and pattern of metastatic spread. Radiology 2003; 226: 527-32). This new review re examines the radiological features of GISTs at presentation and well as their changed imaging features following treatment with Imatinib. PMID- 16361145 TI - Ultrasound of thyroid cancer. AB - The management of thyroid nodules is multi-disciplinary and involves head and neck surgeons, pathologists and radiologists. Ultrasound is easy to perform, widely available, does not involve ionizing radiation and is readily combined with fine needle aspiration cytology (FNAC). It is therefore an ideal investigation of choice for evaluating thyroid nodules. It evaluates specific features that help in identifying the nature of the nodule and FNAC helps in diagnostic accuracy. In addition, following treatment for thyroid cancer ultrasound provides a safe tool for disease surveillance. This paper discusses the role of ultrasound in the management of patients with thyroid cancer. PMID- 16361146 TI - Imaging of lumps and bumps in the nose: a review of sinonasal tumours. AB - Sinonasal disease is one of the most common clinical head and neck pathologies. The majority of sinonasal pathology is inflammatory with neoplasms comprising approximately 3% of all head and neck tumours. Although sinus tumours are rare, they portend a poor prognosis, often due to advanced disease at diagnosis. Like most neoplasms, early detection improves prognosis, therefore clinicians and radiologists should be aware of features separating tumours from inflammatory sinus disease. This article reviews the anatomy, clinical features, imaging findings, treatment and histopathology of selected sinonasal tumours. Benign neoplasms reviewed include osteoma, inverting papilloma, and juvenile nasal angiofibroma. Malignant neoplasms reviewed include squamous cell carcinoma, the minor salivary gland tumour, adenoid cystic carcinoma, adenocarcinoma, melanoma, lymphoma, and olfactory neuroblastoma (esthesioneuroblastoma). PMID- 16361147 TI - Controversies in imaging of hepatocellular carcinoma: multidetector CT (MDCT). AB - Primary hepatocellular carcinoma (HCC) is a significant tumor worldwide and represents the most common primary hepatic neoplasm. Staging criteria are important for appreciation of timely work up of these neoplasms in contradiction with surgical colleagues. This article demonstrates the appearance of HCC on multiphasic, multidetector CT (MDCT) and relates these findings to current staging criteria. The variable appearance on different planes of contrast is critical to appreciate in staging this neoplasm. The hypervascular nature of the primary tumor makes MDCT and three-phase imaging a critical feature in the detection and characterization of this tumor. This is especially critical in the patients who are candidates for surgical resection. Additionally, MDCT has allowed arterial phase imaging to define the vascular supply of the tumor. An accurate representation of the size and number of lesions is critical in not only the initial staging but also the follow-up of hepatocellular carcinoma. The post treatment features including the appearance post-surgically and after radiofrequency ablation can be well appreciated on MDCT. PMID- 16361148 TI - [Rapid detection of Streptococcus pneumoniae and Haemophilus influenzae with DNA probes]. AB - OBJECTIVE: To establish a method for rapid molecular detection of Streptococcus pneumoniae and Haemophilus influenzae in the early stage of infection. METHODS: Specific DNA probes for Streptococcus pneumoniae and Haemophilus influenzae and 16 S rRNA universal probe of bacteria were synthesized by polymerase chain reaction (PCR) and labeled with biotin. The DNA of the bacteria, virus and fungi were hybridized with these probes respectively prior to application for examination of the clinical samples. RESULTS: The DNA probes of 263, 351, and 370 bp were amplified by PCR. Streptococcus pneumoniae and Haemophilus influenzae reacted only with their corresponding probes, and no cross reaction of the bacterial universal probe with virus and fungi was noted. The method could detect bacterial DNA in as small amount as 1 ng. Of the 100 sputum specimens, 11 were found to be positive for Streptococcus pneumoniae and 8 for Haemophilus influenzae, with a positivity rate greater than that by sputum culture. CONCLUSION: DNA probe detection is simple, rapid, and specific for clinical examination of Streptococcus pneumoniae and Haemophilus influenzae. PMID- 16361149 TI - [Effects of ambulatory labor analgesia on fetal oxygen saturation]. AB - OBJECTIVE: To observe the effect of ambulatory labor analgesia on fetal oxygen saturation during labor. METHODS: Sixty parturients with uncomplicated term pregnancies were equally divided, according to their personal preference, into ambulatory analgesia group receiving combined spinal-epidural labor analgesia and control group without analgesia administration. All the parturients with ambulatory labor analgesia received sufentanil and ropivacaine administration. Fetal oxygen saturation was monitored continuously during the labor by recording the values every 5 minutes and the mean value was calculated. Umbilical blood was analyzed after fetal delivery and the pH, base excess (BE) value were recorded. RESULTS: In the analgesia group and the control group, the mean values of fetal oxygen saturation was (51.92+/-5.90)% and (50.77+/-6.47)%, respectively, during the first labor stage (P=0.48), and was (44.85+/-6.27)% and (43.12+/-7.34)% during the second labor stage (P=0.32), and the lowest values during the labor was (41.63+/-7.51)% and (40.96+/-7.48)%, respectively (P=0.73), showing no significant differences in the three values between the two groups. CONCLUSION: Ambulatory labor analgesia does not significantly affect fetal oxygen saturation during the entire course of labor. PMID- 16361150 TI - [Establishing a parabiosis animal model of two-way paradigm]. AB - OBJECTIVE: To establish a parabiosis model between allogenic conspecific adult mice to study two-way paradigm. METHODS: Fifty-four female Balb/c mice and 54 male C57BL/6 mice were paired and equally divided into 3 groups, namely group 1 with normal saline (NS) injection, group 2 with injections of spleen cells and cyclophosphamide (CP), and group 3 injected with spleen cells, CP, and cyclosporin A (CsA). The treatments were performed by injecting the spleen cells from one of the mice in a pair into the other via tail vein and vise versa, and two days after the operation, CP (150 mg/kg) was injected intraperitoneally. Intraperitoneal CsA (30 mg/kg daily) injection was given starting from 2 days before till 7 days after the operation. Twelve of the 18 pairs of parabiosis mice in each group were separated after 1 week, and part of the skin were transplanted to each other. The maintenance of parabiosis was observed in the other 6 pairs of parabiosis mice were observed. Mixed lymphocyte reaction (MLR) and delayed-type hypersensitivity (DTH) were observed and studied in the separated mice. RESULT: The duration of parabiosis maintenance and skin survival of the group 3 was significantly longer than those in the other two groups, and group 3 showed suppressed MLR and DTH. CONCLUSION: With the application of immunosuppressants, we have successfully established the two-way paradigm model in mice. PMID- 16361151 TI - [Different patterns of abnormalities in exercise 201TI myocardial scintigraphy and their mechanisms]. AB - OBJECTIVE: To categorize the patterns of abnormalities in exercise (201)TI myocardial scintigraphy and explore the mechanisms. METHODS: Exercise (201)TI myocardial scintigraphy was performed in 203 patients with clinically suspected coronary artery disease, including 74 normotensive patients, 78 hypertensive patients without left ventricle hypertrophy (LVH) and 51 hypertensive patients with LVH. All the patients underwent coronary angiography one month before or after (201)TI myocardial scintigraphy, and the patterns of abnormal findings were categorized as segmental, non-segmental and mixed patterns. Patients with abnormal (201)TI myocardial perfusion and normal coronary angiographic findings were followed up. RESULTS: In hypertensive patients without and with LVH, the ratios of abnormal perfusion in segmental, non-segmental and mixed types were 52/60 and 32/58, 4/60 and 9/58, and 4/60 and 17/58, respectively. The ratios of normal coronary angiography in the 3 types were 17/84, 13/13 and 10/21, respectively. Among the 40 patients followed up, 5 with segmental abnormality and 2 with mixed abnormalities developed large coronary artery disease, which was found in none of the patients with segmental abnormality. CONCLUSIONS: In hypertensive patients with and without LVH, segmental perfusion abnormalities may be attributed to the anatomic and functional stenosis of the large coronary arteries, and the non-segmental abnormal perfusion might be only possible with coronary microvascular diseases. PMID- 16361152 TI - [Apoptosis induced by curcumin and its effect on c-myc and caspase-3 expressions in human melanoma A375 cell line]. AB - OBJECTIVE: To investigate the effect of curcumin on cell apoptosis and c-myc and caspase-3 expressions in human melanoma A375 cell line. METHODS: A375 cells were exposed to curcumin treatment and growth inhibition of the cells was examined by MTT assay. Annexin V/propidium iodide double staining and DNA fragmentation analysis were employed for assay of the cell apoptosis and morphological changes of the cells were observed with inverted microscopy and transmission electron microscopy, respectively. In situ hybridization and SABC immunohistochemistry were performed for detection of the expressions of c-Myc and caspase-3 in the A375 cells. RESULTS: Curcumin inhibited the growth of A375 cells in both time- and concentration-dependent manners. After treatment with 30 micromol/L curcumin for 48 h, apoptotic morphological changes were observed in the cells and an oligonucleosomal DNA ladder was clearly visualized in DNA fragmentation analysis. The apoptotic rates of the cells treated with curcumin at the concentration above 20 micromol/L were significantly higher than that of the control cells. c-myc expression level was decreased whereas caspase-3 expression increased with the increase in curcumin concentrations. CONCLUSION: Curcumin can inhibit the proliferation and induce apoptosis of A375 cells in vitro, and the genes encoding c-myc and caspase-3 may play a role in the process. PMID- 16361153 TI - [Clinical nature of tetrodotoxin intoxication and the measures for emergency rescue]. AB - OBJECTIVE: To define the clinical nature of tetrodotoxin intoxication and explore the measures for its emergency rescue. METHODS: The emergency rescue measures and outcomes of 33 cases tetrodotoxin intoxication were reviewed. RESULTS: No obvious effects were observed in these patients after administration of atropine or application of respiratory stimulant, but implementation of mechanical ventilation and comprehensive life support measures resulted in successful rescue of mild to moderate cases within 10 to 14 h, and recovery of spontaneous respiration was achieved in critical cases in 19-24 h. CONCLUSION: Tetrodotoxin selectively blocks the neuromuscular electrical activity, producing the effect of nondepolarizing curine and leading to failure of the respiratory drive without compromising the respiratory center. Atropine, a M-cholinergic receptor blocker, or central stimulants may fail to produce obvious effects, but comprehensive treatments including maintenance of effective ventilation can bring satisfactory effects. PMID- 16361154 TI - [Expression of COX-2 protein in colorectal carcinoma and the clinical implication]. AB - OBJECTIVE: To investigate the relationship between COX-2 expression and the clinicopathological factors in colorectal carcinoma and assess the prognostic value of COX-2 detection. METHODS: Tissue microarray and immunohistochemistry by SABC method was employed for detecting COX-2 expression in 126 patients with advanced colorectal cancer, and the relationship of COX-2 expression with the clinicopathological features and prognosis of the patients was retrospectively analyzed. RESULTS: The patients were divided into two groups of low and high COX 2 groups according to the grade and extent of COX-2 expression. High COX-2 expression was detected in 32 (25.4%) cases, and low expression in 94 (74.6%) cases. No significant correlation was noted between COX-2 expression and the patients' age, sex, tumor size, tumor location, histological type, lymphatic-node metastasis and Dukes' classification, but high COX-2 expression was strongly correlated with tumor recurrence and especially with blood-borne metastasis (P<0.05). The survival rate without tumor recurrence for high- and low-COX-2 groups was assessed by the Kaplan-Meier method and compared by log-rank test, which revealed significant difference between the two groups (P=0.0067). Multivariate analysis for all patients suggested that among the 8 prognostic factors (age, sex, tumor size, tumor location, histological type, lymphatic-node metastasis, Dukes' stage, and COX-2 expression), Dukes' stage and COX-2 expression was the independent significant factor related to disease-free survival. CONCLUSION: The expression of COX-2 is strongly correlated with recurrence of colorectal cancer, especially with blood-borne metastasis. COX-2 is an independent factor for prognostic evaluation of the patients, and tissue microarray allows rapid, convenient, economic and accurate COX-2 detection for large-scale application. PMID- 16361155 TI - [Analysis of drug resistance spectrum and its mechanism in 1017 clinical bacterial isolates]. AB - OBJECTIVE: To investigate the drug susceptibility of the clinical bacterial isolates to provide evidence for more adequate use of antibiotics. METHODS: Altogether 1017 clinical bacterial isolates were identified by BD Phoenix and tested for resistance against antimicrobial agents by K-B method. WHONET5 was applied for the analysis. RESULTS: The most frequent bacteria detected included P. aeruginosa (19.37%), coagulase-negative Staphylococci (CNS, 17.70%), E. coli (13.27%), S. aureu (SA, 12.09%), E. faecalis (11.8%), and K. pneumoniae (7.57%). In gram-negative isolates, the susceptibility rate of imipenem was 81.5%, and that of ceftazidime was above 70%. The incidences of E.coli and K. pneumoniae isolates producing extended spectrum beta-lactamase (ESBLs) were 34.8% and 45.5% respectively. In gram-positive isolates, the susceptibility rates of vancomycin and Teicoplanin were 98.8% and 100.0% respectively, and those of furazolidone, imipenem, amikacin, piperacillin/ tazobactam were above 70%. The oxacillin resistant rates of CNS and SA were 78.2% and 46.8%. CONCLUSION: The 1017 clinical bacterial isolates are characterized by high ratio of ESBL production and oxacillin resistance, suggesting the importance of adequate use of antimicrobial agents and effective control measures for reducing the drug resistance and preventing the spread of multi drug- resistant bacteria. PMID- 16361156 TI - [Therapeutic effect of TAX combined with Herceptin or epirubicin against breast cancer positive for Her-2/neu]. AB - OBJECTIVE: To compare the therapeutic effects of biochemotherapy regimen with Herceptin plus taxol (TAX) and the chemotherapy regimen with epirubicin plus TAX against Her-2/neu-positive breast cancer and observe the changes in serum tumor markers in patients receiving biochemotherapy. METHODS: Seventy-three patients with advanced breast cancer positive for Her-2/neu as revealed by immunohistochemistry were divided into the study group (n=32) to receive treatment with the regimen of Herceptin plus TAX and control group (n=41) with the regimen of epirubicin(EPI) plus TAX. The therapeutic effects of the regimens were observed and in the study group, the relationship of the therapeutic effect with Her-2/neu positivity and changes in serum tumor markers were examined. RESULTS: The objective response rate and clinical benefit response rate were obviously higher in the study group than in the control group. In the study group, the clinical response rate of patients with positive immunostaining for Her-2/neu of grades 1+, 2+ and 3+ were 0%, 44.4% and 63.6%, respectively, as compared with those in the control group of 8.3%, 36.4%, and 38.9%, respectively, and the treatment resulted in lowered levels of serum tumor markers without significant changes in CA153, tps and CEA (P<0.05) after treatment, but CA125 showed no significant difference (P>0.05). CONCLUSIONS: In patients with advanced breast cancer with positive immunostaining for Her-2/neu of grade 3+, the regimen of Herceptin plus TAX can be more effective than the chemotherapeutic regimen of EPI plus TAX. Patients with Her-2/neu (+++) benefit more from the treatment than those with Her-2/neu (++). Serum CEA, CA153 and TPS levels also possess some value in evaluating the therapeutic effects of the regimens. PMID- 16361157 TI - [bcl-2 expression in small cell lung cancer: a mechanism for apoptosis antagonism and immune evasion]. AB - OBJECTIVE: To observe bcl-2 expression and its correlation with apoptosis antagonism and immune evasion of small cell lung cancer cell line and explore the application of bcl-2 antisense thio-oligonucleotide (bcl-2 SON) in gene therapy for lung cancer. METHODS: Western blotting was performed to detect bcl-2 expression in NCI-H69 cell line expressing Bcl-2 protein treated with bcl-2 SON. The treated NCI-H69 cells, along with the cells without bcl-2 SON treatment and NCI-H82 cells that did not express Bcl-2 protein, were respectively co-cultured with tumor-infiltrating lymphocytes (TILs) isolated from fresh tumor samples, and the apoptosis of the lung cancer cells was assessed by JAM assay. RESULTS: Western blotting revealed obvious inhibition of bcl-2 expression in NCI-H69 cells in response to bcl-2 SON treatment. JAM assay showed that the apoptosis of bcl-2 SON-treated H69 cells and NCI-H82 cells increased with the elevation of TIL ratio in the co-culture, while such changes were not observed in NCI-H69 cells without bcl-2 SON treatment. CONCLUSION: Small cell lung cancer cells expressing bcl-2 may antagonize the antitumor immune attack in the host, and bcl-2 SON may provide an effective alternative in gene therapy for small cell lung cancer. PMID- 16361159 TI - [Application of flexible bronchoscopy in postoperative management of lung transplantation]. AB - OBJECTIVE: To investigate the application of flexible bronchoscopy in the postoperative management of lung transplantation. METHODS: Four male patients with end-stage chronic obstructive pulmonary disease received examination with bronchoscope following the unilateral lung transplant for management of airway complications. One patient developed stenosis at tracheal anastomosis 2 months after transplantation and underwent balloon dilatation and endobronchial metallic stent placement. RESULTS: The 4 patients showed good recovery after transplantation with obviously improved pulmonary function and quality of life. CONCLUSION: Flexible bronchoscopy provides a valuable means for postoperative management of lung transplantation. PMID- 16361158 TI - [Therapeutic effects of anterolateral decompression and fixation on thoracolumbar fractures complicated with incomplete paraplegia]. AB - OBJECTIVE: To investigate the clinical effects of anterolateral decompression and fixation on thoracolumbar fractures complicated with incomplete paraplegia. METHODS: Thirty-six patients with thoracolumbar fractures complicated with incomplete paraplegia were treated with anterolateral decompression and fixation. RESULTS: The patients were followed up for an average of 18 months, which showed satisfactory recovery of the intervertebral space height and thoracolumbar vertebral curvature. The average Cobb's angle, spinal canal index and Frankel were improved remarkably, and none of the patients developed such complications as break or mobilization of the plate screw. CONCLUSION: Anterolateral decompression and fixation can directly and completely decompress the vertebral canal, promote the functional recovery of the spinal nerves and reconstruct the alignment of the spine as an ideal approach for treatment of thoracolumbar fractures with obvious spinal canal-occupying lesions or severe kyphos complicated with incomplete paraplegia. PMID- 16361160 TI - [Surgical treatment of anterior circulation aneurysm via pterion keyhole approach]. AB - OBJECTIVE: To investigate the feasibility of pterion keyhole approach with minimal invasion for treatment of the anterior circulation aneurysm. METHOD: Aneurysm clipping through the pterion keyhole approach was performed in patients with anterior circulation aneurysms, including 9 with posterior communication artery aneurysms, 3 with middle cerebral artery aneurysms and 6 with anterior communication artery aneurysms, who were in stages I to III according to Hunt Hess scale. RESULT: All the aneurysms were clipped successfully. One patient with a left posterior communication artery aneurysm developed transient sensory aphasia and motor aphasia after surgery due to intraoperative aneurysm rupture. No facial paralysis occurred due to damage of the facial nerve. CONCLUSION: The pterion keyhole approach is a very useful surgical approach for treatment of anterior circulation aneurysms on the basis of cautious determination of indications and careful operation planning. PMID- 16361161 TI - [Determination of amygdalin content in Semen Armeniacae Amarum by high performance liquid chromatography with evaporative light-scattering detection]. AB - High-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD) was employed for determination of amygdalin content in Semen Armeniacae Amarum. The detection was performed with the column of Hypersil ODS (4.6 mm x 250 mm, 5 microm) and column temperature of 25 degrees C. The mobile phase was methanol-water (70:30) with flow rate of 0.5 ml/min. Evaporative light-scattering detector was used and the drift tube temperature was set at 98 degrees Celsius with the gas flow rate of 3.2 L/min. A standard curve was generated, which was linear in the range of 1.0-15.1 microg for amygdalin content (r=0.999 9). The average recovery of amygdalin was 99.0% with RSD of 2.9% (n=5). Besides simplicity and rapidness, the method yields accurate and reproducible results and can therefore be used in the quality control of Semen Armeniacae Amarum. PMID- 16361162 TI - [Magnetic resonance imaging and radiography for diagnosis of lower limb osteosarcoma: a comparative study]. AB - OBJECTIVE: To investigate the diagnostic value of magnetic resonance imaging (MRI) for lower limb osteosarcoma by comparing the findings between X-ray and MRI. METHODS: Fifteen surgically or pathologically confirmed patients with lower limb osteosarcoma (11 males and 4 females) underwent both plain and enhanced MRI (with CT scanning in 4 cases) in addition to radiography, and the findings were compared. RESULTS: The diagnostic accuracy of both MRI and X-ray film was 100%. Compared with X-ray, MRI was capable of clear display of the lesion extent, perpendicular periosteal reaction, the two layers of Codman's triangle and the skip lesions. CONCLUSION: Plain X-ray film provides a convenient and practical diagnostic modality for lower limb osteosarcoma, but MRI has better performance in clearer display of the tumor range and identification of early-stage lesions. PMID- 16361163 TI - [Effect of primary percutaneous coronary intervention on plasma B-type natriuretic peptide in patients with acute myocardial infarction]. AB - OBJECTIVE: To observe the effect of primary percutaneous coronary intervention (PCI) on plasma B-type natriuretc peptide (BNP) in patients with acute myocardial infarction (AMI). METHODS: Thirty-eight patients with AMI were divided into two groups for PCI (n=26) and conventional treatment (n=12). The plasma BNP levels were measured by fluorescence immunoassay (FIA) in these patients immediately, 24 h, 7 d, and 30 days after admission, and the infarct-related coronary arteries (IRA) were treated only with emergency interventional therapy in PCI group. RESULTS: BNP of the patients in the PCI and conventional treatment group B increased immediately and 24 h after admission, but there was no significant difference between the two groups (243.74+/-75.68 vs 228.65+/-82.32 and 283.42+/ 88.66 vs 275.48+/-89.67, P>0.05). BNP in PCI group decreased but that in conventional treatment group increased 7 days after admission, showing significant difference between them (203.63+/-59.42 vs 388.74+/-108.52, P<0.05 ). BNP remained significantly lower in the PCI group than in the other group 30 days after admission (96.31+/-43.22 vs 237.66+/-75.48, P<0.01). Emergency PCI for different IRA resulted in the significant difference in BNP between the patients, and intervention of the left anterior descending artery (LAD) resulted in more obvious BNP reduction in comparison with that due to interventional of the right coronary artery (RCA) and left circumflex coronary artery (LCX). The changes in left ventricular ejection fraction (LVEF) and left ventricular end-diastolic dimension (LVEDD) were correlated to the changes of BNP. The mean BNP in the 3 fatal cases was nearly 10-fold higher than the normal level. CONCLUSIONS: BNP of AMI patients decreases on days 7 and 30 after reperfusion therapy with primary PCI, and the reduction can be more obvious 30 days after admission. PMID- 16361164 TI - [Effect of Jingjielianqiao decoction on healing of skin ulcer in rats]. AB - OBJECTIVE: To study the pharmacodynamics of Jingjielianqiao decoction in promoting the healing of skin ulcer in mice. METHODS: Mouse models of skin ulcer were established by inducing skin trauma and scald, respectively. The skin ulcer of the mice was treated with Jingjielianqiao decoction or recombinant human epithelial growth factor (rhEGF), and the ulcer area after treatment was compared between the two groups and with their respective self control. Inflammatory cell infiltration in the ulcer was observed under the microscope. RESULTS: Compared with the self-control group, the skin ulcer area of mice treated with Jingjielianqiao decoction was reduced obviously (P<0.001), and the inflammatory cell infiltration also alleviated (P<0.001). Jingjielianqiao decoction produced significantly better therapeutic effect than rhEGF. CONCLUSION: Jingjielianqiao decoction can obviously improve the healing of skin ulcer in mice with better effect than rhEGF. PMID- 16361165 TI - [Value of multislice spiral CT scanning and three-dimensional reconstruction in selecting optimal length and shape of mandible internal fixation material]. AB - OBJECTIVE: To investigate the value of multislice spiral CT (MSCT) scanning and three-dimensional reconstruction in selecting optimal length and shape of mandible internal fixation material. METHODS: Three-dimensional reconstruction of the mandible was performed in 5 cases by MSCT scanning and the length of the mandible for operation was measured. Some isometric points were chosen on the mandibular curve and the distance from these points to the two ends of the curve was measured to depict the mandibular curve using a self-designed skeletal curve depicting software. The optimal length and shape of the material for internal fixation was then determined. RESULTS: After removal of the soft tissues and other irrelevant bones from the reconstructed three-dimensional mandible with clear view from any directions, the length of the mandible was accurately measured with the software. The physiological curve of the mandible was generated after determination of the distances from the chosen points on the mandibular curve and to the two ends of the curve for precise determination of the length and shape of the material for internal fixation. CONCLUSION: Three-dimensional mandible reconstruction with MSCT and measurement facilitates the determination of the optimal length and shape of the internal fixation material to enhance the attachment between the material and the bone surface and consequently the therapeutic effects. PMID- 16361166 TI - [Comparison of the block characteristics of levobupivacaine vs bupivacaine for unilateral spinal block]. AB - OBJECTIVE: To compare the block characteristics of levobupivacaine and bupivacaine for hypobaric unilateral spinal block. METHODS: Sixty patients undergoing hypobaric unilateral spinal block for lower extremity operations were randomized into 2 equal groups. In one group the patients were asked to lie on the uncompromised side for slow injection of hypobaric bupivacaine solution (0.75% 6 mg, diluted to 2 ml with distilled water) at the rate of 1 ml/15 s through a 27-gauge needle. The patients maintained their position for 15 min before shifting to a supine position. In the other group levobupivacaine was used instead. RESULTS: Statistical study showed that the block characteristics were of no significant difference between the 2 groups (P>0.05) except that the duration of motor block was shorter in levobupivacaine group than in bupivacaine group (P<0.05). Blood pressure or heart rate showed no significant difference between the two groups during operation. CONCLUSION: The block characteristics of hypobaric unilateral spinal block with levobupivacaine or bupivacaine are similar and their effect on circulatory function is slight. Levobupivacaine provides an option in case bupivacaine is not available. PMID- 16361167 TI - [Application of Helena SAS-3 full automatic hemoglobin agarose electrophoresis]. AB - OBJECTIVE: To assess the value of Halena SAS-3 full automatic hemoglobin agarose electrophoresis in the diagnosis of thalassemia. METHODS: Hemoglobin A and A2 was detected by agarose electrophoresis in 200 blood samples collected from 170 healthy subjects, 2 patients with beta-thalassemia, 12 with acute lymphoblastic leukemia, 12 acute myelocytic leukemia and with 4 non-Hodgkin lymphoma. RESULTS: The Mean+/-SD of hemoglobin A in the healthy subjects and patients with beta thalassemia, acute lymphoblastic leukemia, acute myelocytic leukemia and non Hodgkin lymphoma was 97.55+/-0.51, 97.01+/-0.329, 97.42+/-0.57, 97.44+/-0.55, and that of A2 was 2.44+/-0.51, 2.99+/-0.32, 2.84+/-0.28, 2.55+/-0.55, respectively. The values of electrophoresed hemoglobin F of the 2 patients with beta thalassemia were higher than those of the normal adults. The within-run coefficient of variation (CV) and between-run CV was 0.17 and 7.76 for hemoglobin A, and 0.16 and 7.14 for hemoglobin A2, respectively. CONCLUSIONS: Halena SAS-3 full automatic agarose electrophoresis system possesses strong resolving power in detecting trace abnormal hemoglobin and accurately identifies unusual evaluation of hemoglobin A2 and F to facilitate the diagnosis of thalassemia. PMID- 16361168 TI - [Value of 18F-fluorodeoxyglucose positron emission tomography in three dimensional conformal radiotherapy for locally persistent or recurrent nasopharyngeal carcinoma]. AB - OBJECTIVE: To explore the clinical value of (18)F-fluorodeoxyglucose positron emission tomography ((18)FDG-PET) in three-dimensional conformal radiotherapy (3D CRT) for locally residual and recurrent nasopharyngeal carcinoma (NPC). METHODS: Between July 1998 and October 2000, 36 patients with recurrent or residual NPC confirmed by biopsy after initial radiotherapy underwent treatment with 3D-CRT. CT simulation was performed and (18)FDG-PET carried out for delineating the gross tumor volume (GTV) in all the patients. The radiotherapy was designed by 3D treatment planning system and dose-volume histogram (DVH) was used to verify and optimize the radiation plan. Acute and late radiation injuries, survival rates and GTV delineated by CT and (18)FDG-PET were observed in all the cases after the treatment. RESULTS: In comparison with CT simulation, (18)FDG-PET delineated similar GTV in 36.1% (13/36) cases, larger and smaller GTV in 16.7% (6/36) and 47.2% (17/36) of the cases, respectively. After 3D-CRT, 97.2%, 75.0%, and 58.3% of the patients were free of evidence of 1-, 2-, and 3-year local failure, respectively, and the 1-, 2-, and 3-year survival rate of the patients were 94.4%, 86.1%, and 72.0%. The 1-, 2-, 3-year tumor-free survival rates were 85.7%, 76.6%, and 56.1% respectively. Acute xerostomia, nasopharyngitis and oropharyngitis were the most common complications in relation to the radiation, but all within acceptable range of grades 1 or 2. The lesions resulting from the radiation included hearing loss, trismus, dysphagia, chronic sinusitis, and cranial neuropathy, with the incidences of 5.6% (2/36), 25.0% (9/36), 8.3% (3/36), 33.3% (12/36), and 13.9% (5/36) respectively. CONCLUSIONS: 3D-CRT combined with (18)FDG-PET for delineation of GTV is effective and feasible in the treatment of locally residual or recurrent nasopharyngeal carcinoma after initial radiotherapy, producing good responses without serious complications. PMID- 16361169 TI - [A clinical study of lobectomy with minimally invasive incision]. AB - OBJECTIVE: To study the application of small thoracic incision for lobectomy with effects comparable to that of conventional incision. METHODS: A total of 386 patients with lung tumors were prospectively randomized into thoracic small incision group (n=184; average length of incision, 12.7 cm) and standard thoracic incision group (control group, n=152). The incision length, operation time, intra operative bleeding volume, postoperative drainage volume, transfusion, hemostatic dosage, ambulation time, hospital stay and major complications were compared between the two groups. RESULTS: All of the indices in small incision group were significantly better than those of the control group (P<0.01) except the operation time (P>0.05). No severe complications occurred in small incision group, and in 4 cases conventional thoracotomy was performed instead due to adhesion or giant tumors. CONCLUSIONS: Muscle-sparing thoracic small incision can achieve excellent clinical result and has such advantages as less invasiveness, reduced bleeding volume, pain and hospital stay, less complications, and better postoperative appearance. This procedure meets the requirements of lobectomy and provides an alternative for routine lobectomy for pulmonary tumors. PMID- 16361170 TI - [Clinical analysis of the pathogenesis of mycoplasma pneumonia in children]. AB - OBJECTIVE: To study the clinical features and immune-mediated pathogenesis of mycoplasma pneumonia (MP) in children. METHODS: The DNA of mycoplasma in nasopharyngeal swabs was detected by PCR in 485 children with respiratory tract infection, and serum circulating immune complex (CIC), IgG, IgM, MP IgM, and CH50 were measured on the same day in 45 children with pneumonia. RESULTS: Significant difference was noted between the results of mycoplasma DNA PCR detection and those of serum IgM detection. The positivity rate of CIC was 44% in children with MP whereas that in the control group was only 10% (P<0.01). The levels of serum IgG and IgM in MP group was significantly higher and serum CH50 lower than those of the control group (P<0.05). CONCLUSION: Mycoplasma DNA detection facilitates early diagnosis and treatment of MP, whose pathogenesis in children may be involved in immune-mediated mechanism. PMID- 16361171 TI - [Causes of failure in choledochoscopic diagnosis and removal of gallstone]. AB - OBJECTIVE: To discuss the causes for failure in diagnosis and removal of gallstone using choledochoscope. METHOD: A retrospective analysis was conducted in 714 cases of gallstone treated with choledochoscopic operations. RESULTS: Residual stones in the bile duct were found in 132 (18.5%) cases after the operation. In 54 cases the bile duct stones failed to be completely removed even before the closure. By T-tube cholangiography and choledochoscopy after the operation, 78 cases were found to have residual bile duct stones, among them 29 were complicated with intrahepatic bile duct and bile duct opening stenosis, 37 with intrahepatic bile stone and bile duct variation, and 12 with choledochus inferior segment stenosis and stone formation. CONCLUSION: Due attention should be paid during the choledochoscopic operation to bile duct variation, bile duct opening stenosis or choledochus inferior segment stenosis, which should be carefully inspected by segments in succession to minimize the possibility of residual stones. PMID- 16361172 TI - [Clinical observation of simple rhinologic headache due to rhinal structural abnormalities]. AB - OBJECTIVE: To analyze the causes and evaluate the therapies for simple rhinologic headache resulting from rhinal structural abnormalities. METHOD: A retrospective analysis of 62 cases of simple rhinologic headache was conducted. All the patients were examined by high-resolution CT and nasal endoscopy to identify the anatomic structural abnormality in the nasal cavity. After establishment of the diagnosis, the nasal abnormalities were corrected by nasal endoscopic surgery. RESULT: Of all the cases, 46 were cured, 12 responded favorably to the treatment, and 4 failed to respond possibly due to multiple causes of the headache and misdiagnosis. CONCLUSION: High-resolution CT combined with nasal endoscopy may improve the diagnosis of simple rhinologic headache and nasal endoscopic surgery may serve as an effective modality for its treatment by correcting the nasal structural abnormalities. PMID- 16361173 TI - [Effects of penehyclidine hydrochloride or atropine combined with neostigmine for antagonizing residual neuromuscular block on patient's hemodynamics]. AB - OBJECTIVE: To investigate the effects of penehyclidine hydrochloride(PH)/atropine combined with neostigmine for antagonizing residual neuromuscular block on the hemodynamics. METHODS: Eighty patients with elective upper abdominal surgery were randomized equally into PH group and atropine group. Five minutes after the completion of surgery, PH 0.02 mg/kg (in PH group) or atropine 0.02 mg/kg (in atropine group) in combination with neostigime 0.03 mg/kg were given intravenously to reverse the residual neuromuscular block. The heart rate (HR), mean arterial pressure (MAP) and end-tidal CO(2) pressure (P(ET)CO(2)) were recorded 5 min before anesthesia induction, 1 min before injection and 2, 5, 10, and 15 min after injection, respectively. RESULTS: During the investigation, HR in PH group did not undergo conspicuous changes (P>0.05). HR after the injection was markedly faster than that before the injection in atropine group (P<0.01) and did not recover till 15 min after the injection (P>0.05). MAP and P(ET)CO(2) showed no evident changes and no significant difference was observed between the two groups during the investigation (P>0.05). CONCLUSION: Compared with atropine, PH does not obviously affect HR and BP, but atropine may accelerate HR. PMID- 16361174 TI - [Therapeutic effect of phacoemulsification for closed-angle glaucoma]. AB - OBJECTIVE: To investigate the therapeutic effect of phacoemulsification for primary closed-angle glaucoma complicated with cataract. METHODS: Phacoemulsification with posterior chamber foldable intraocular lens implantation was performed in 55 cases (58 eyes) of acute or chronic primary closed-angle glaucoma complicated with cataract. The changes of visual acuity, intraocular pressure, central anterior chamber depth and anterior chamber angle were observed after operation and the patients were followed up for 6 months. RESULTS: Compared with those before operation, the postoperative best corrected visual acuity and anterior chamber depth were improved, the intraocular pressure was reduced, and the closed chamber angle was partially reopened. CONCLUSION: Phacoemulsification with posterior chamber foldable intraocular lens implantation not only improves the visual acuity but also controls the intraocular pressure, and can be a valuable method for treating primary closed-angle glaucoma with cataract. PMID- 16361175 TI - [Clinical analysis of 73 cases of hypoglycemia with brain dysfunctions]. AB - OBJECTIVE: To identify the clinical features and pathogenesis of hypoglycemia with brain dysfunctions as the main manifestations. METHOD: A retrospective analysis of 73 cases with brain dysfunctions caused by hypoglycemia was performed. RESULT: Hypoglycemic brain dysfunctions were mainly caused by poor control of the dosage of hypoglycemic agents in diabetic patients, and the major clinical manifestations included coma, hemiparalysis, epilepsy and mental disorders. The pathogenesis was complicated, possibly related to a variety of factors such as rapid declination of blood sugar level, aging, cerebral arteriosclerosis, and hypoglycemic cerebral vasospasm or selective nerve injury. CONCLUSIONS: A blood sugar test should be performed for any patients with brain dysfunctions of unidentified causes for early diagnosis and treatment. In addition, diabetic patients should take hypoglycemic agent adequately. Regular monitoring of blood sugar level is key to the prevention of hypoglycemia. PMID- 16361176 TI - [Summary of Guangdong provincial seminar on avian influenza and influenza]. AB - On 8th November 2005, an academic seminar on avian influenza and influenza in Guangdong Province was held by Guangdong Society of Tropical Medicine and the Epidemiology Committee of the Guangdong Preventive Medicine Society in Southern Medical University, addressing the current problems in epidemics of avian influenza. The specialists attending the conference arrived at the common consideration that at present, the avian influenza virus H5N1 has not the capacity to trigger an pandemic in human population, but scattered cases had been reported to increase the suspicions of H5N1 virus transmission between humans. Due attention should be paid to the tendency of expansion of the host range and epidemic area, and the possibility of disastrous influenza pandemic among human populations persists, for which rational consideration is called for, and the role of specialists should be fully recognized who are endeavoring to examine the possible scale of influenza occurrence and devise strategy to deal with the epidemic in Guangdong province according to the practical situation in China. Increased funds and investment in scientific research on avian influenza is urged for influenza prediction and surveillance, rapid and early diagnostic assays, understanding of virus variation, mechanism of H5N1 virus adaptation to human hosts, effective medicines and vaccines for prevention and therapy of avian influenza. Laboratory bio-safety control should be enforced to prevent infections originated from laboratories. The specialists appeal that the media report the news objectively and issue the public warnings against avian influenza after consulting specialists, so as to avoid unnecessary social panic. PMID- 16361177 TI - [Application of laminar air flow techniques in burn treatment]. AB - OBJECTIVE: To evaluate the value of laminar flow in the treatment of burns. METHODS: The air in the laminar flow chamber and the wound tissues of the patients were sampled for bacterial detection. The number and stains of bacterial colony from different classes of laminar air flow chambers at different time points were inspected and compared. RESULTS: The bacterial number was 0 in the laminar flow chamber of 1000 grade, which was obviously different from that in the public area. The mortality was obviously decreased in the laminar air flow chamber with shorter treatment time and hospitalization. No wound infection occurred and the wounds healed smoothly in all these patients. CONCLUSION: The application of laminar air flow can be helpful for the treatment of severe burns. PMID- 16361178 TI - Schoolchildren's food consumption and dietary intake during the dry season in north-west Namibia. AB - The diet of school-aged children in north-west Namibia was investigated. Data on food consumption were collected using the 24-h recall interview method. The field study was conducted during the dry season. The subjects were schoolchildren aged between 8 and 15 years living in either a small town (n=43) or a rural area (n=10). In town, the main sources of energy were maize porridge and wheat bread, and for the rural children, sour milk and maize. The consumption of vegetables, fruit and legumes was very low in both groups. The mean intakes of energy were 6.7 MJ/day for children living in town and 4.7 MJ/day for rural subjects. The intakes of vitamin A, vitamin C and folate were inadequate in both groups. Our findings show that during the dry season, the diets of Namibian school-aged children are monotonous. PMID- 16361179 TI - Antinutritional factors and in vitro protein digestibility of improved haricot bean (Phaseolus vulgaris L.) varieties grown in Ethiopia. AB - The antinutrient (raffinose oligosaccharides, tannins, phytic acid and trypsin inhibitors) composition and in vitro protein digestibility of eight improved varieties of Phaseolus vulgaris grown in Ethiopia were determined. Stachyose was the predominant alpha-galactosides in all haricot bean samples. Raffinose was also present in significant quantities but verbascose, glucose and fructose were not detected at all in the samples. The concentrations observed for the protein digestibility and antinutritional factors, varied significantly (P<0.05) between varieties investigated in this study. Mean values for protein digestibility ranged from 80.66% (in Roba variety) to 65.64% (in Beshbesh variety). Mean values for raffinose, stachyose, sucrose, trypsin inhibitors, tannins and phytic acid were 3.14 mg/g, 14.86 mg/g, 24.22 mg/g, 20.68 TUIx10(3)/g, 17.44 mg, catechin equivalents/g and 20.54 mg/g respectively. Statistical analyses of data revealed that antinutritional factors and protein digestibility were influenced by variety (genotype). Relationships between antinutritional factors and protein digestibility were also observed. The possibility of selecting varieties to be used for large-scale cultivation in Ethiopia on the basis of these data is discussed. Among the improved varieties studied, Roba, Redwolaita, Mexican and Awash were found to be the best food and export type of haricot beans in the Ethiopian context, because of their higher protein digestibility, lower antinutrtional factors and other beneficial nutritional parameters. Roba variety can be used by local food processors for the production of value-added bean-based products especially to combat the problem of protein energy malnutrition and related diseases which are very common in developing countries. PMID- 16361180 TI - Degradation kinetics of folic acid in cowpea (Vigna catjang L.) during cooking. AB - Kinetics of folic acid degradation in cowpea (Vigna catjang L.) as well as in pure folic acid solutions at initial concentrations present in cowpea over a temperature range of 50-120 degrees C (isothermal conditions) followed first order reaction kinetics, where the rate constant increased with an increase in temperature. The temperature dependence of degradation was adequately modeled by Arrhenius equation. The degradation was also evaluated in normal open pan cooking, pressure-cooking and a newly developed and patented fuel-efficient Eco Cooker (non-isothermal heating process). A mathematical model was developed using the isothermal kinetic parameters obtained to predict the losses of folic acid from the time-temperature data of the non-isothermal heat processing method. The results obtained indicate the folic acid degradation is of a similar order of magnitude in all the methods of cooking. PMID- 16361181 TI - Plant foods in the management of diabetes mellitus: spices as beneficial antidiabetic food adjuncts. AB - Diet has been recognized as a corner stone in the management of diabetes mellitus. Spices are the common dietary adjuncts that contribute to the taste and flavour of foods. Besides, spices are also known to exert several beneficial physiological effects including the antidiabetic influence. This review considers all the available information from animal experimentation as well as clinical trials where spices, their extracts or their active principles were examined for treatment of diabetes. Among the spices, fenugreek seeds (Trigonella foenumgraecum), garlic (Allium sativum), onion (Allium cepa), and turmeric (Curcuma longa) have been experimentally documented to possess antidiabetic potential. In a limited number of studies, cumin seeds (Cuminum cyminum), ginger (Zingiber officinale), mustard (Brassica nigra), curry leaves (Murraya koenigii) and coriander (Coriandrum sativum) have been reported to be hypoglycaemic. PMID- 16361182 TI - In vitro intestinal transport and antihypertensive activity of ACE inhibitory pea and whey digests. AB - Angiotensin I converting enzyme (ACE) inhibitory peptides cause an antihypertensive effect if they reach the systemic circulation. This was investigated for the high ACE inhibitory activity present in peas and whey in vitro gastrointestinal digests. The samples retained high ACE inhibitory activity when incubated in Caco-2 homogenates or rat intestinal acetone powder, both sources of small intestine peptidases. Only little ACE inhibitory activity was transported through Caco-2 cell monolayers in 1 h. As the Caco-2 model is tighter than intestinal mammalian tissue, sufficient absorption of these peptides might occur in vivo. After intravenous administration of 50 mg protein kg(-1) BW in spontaneously hypertensive rats (SHR), pea digest exerted a transient, but strong antihypertensive effect of 44.4 mmHg. Whey digest exerted no effect at this dose. These results suggest that pea digest could be a promising source of ACE inhibitory peptides for use in the prevention and treatment of hypertension. PMID- 16361183 TI - Differences in reported winter and summer dietary intakes in young adults in Spain. AB - Representative data of usual food consumption are necessary in order to establish accurate conclusions on the relationship between diet and disease. In most studies data are collected at one particular time of the year. The aim of this study was to determine the influence of season of the year on dietary intake estimates. Three hundred and three people aged 19-40 years from northwest Spain recorded their intake twice (in winter and summer) using the seven non consecutive-day dietary record method. A total of 247 foods (arranged into 16 major food groups) and 27 nutrients were determined. Most fruits and vegetables, as well as some milk products showed significant seasonal differences. Daily total food consumption amount (g) was higher in winter than summer in males. Higher average intakes of energy (men) and a large number of nutrients (both sexes) were observed in winter. This seasonal influence on dietary intake estimates implies significant differences in fractions of Spanish recommendations covered (lower in summer) for energy (men) and most nutrients (both sexes). As expressed in amount/MJ (densities), most nutrients showed higher values in winter in women. Data in our study suggest that the substantial variations of nutrient intakes throughout the course of the year mainly depend on food (energy) amount variations (quantitative diet changes) in males, while changes in densities of nutrients (qualitative diet changes) are mainly responsible for dietary variations in females. Results in the present study suggest that considering the season of the year it will enhance the results of dietary surveys and ensure the accuracy of studies that link diet with disease. PMID- 16361184 TI - Physicochemical, nutritional and sensory quality of stirred 'dwarf' golden apple (Spondias cytherea Sonn) yoghurts. AB - The dwarf golden apple (Spondias cytherea Sonn) is an exotic fruit which is mainly consumed in its fresh form but could be processed. The objectives of the study were to investigate the effects of adding golden apple nectar on the physicochemical and sensory quality of stirred yoghurts from cow's milk. Yoghurts with 15% and 20% golden apple nectar were more (P>0.01) liked than the control (0% nectar) yoghurt in all sensory attributes. The appearance and body attribute differed (P<0.01) between yoghurts with 15% and 20% nectar. The overall sensory quality of these yoghurts was rated very good to excellent. There were significant (P>0.05) changes, in pH, lactic acid, consistency, colour, lactic acid bacteria and yeasts and moulds on storage at 4 degrees C for 4 weeks. By week 4 of storage, yoghurts developed a buttery smell and were less dark and yellow. A 226 g yoghurt serving provided an excellent source of phosphorus and was good in protein. PMID- 16361185 TI - The management of posterior polar cataract: the role of patching and grading. AB - BACKGROUND: The milder form of posterior polar cataract is often discovered late in preschool children if secondary symptoms like strabismus are not present. The question addressed in this paper is: do such children still profit from orthoptic and surgical treatment? PATIENTS AND METHODS: This was a retrospective study of the data from 13 patients, median age 4 years (range: 1 month to 12 years), with a mean follow-up of 4 years. Four changes to the red reflex were introduced. RESULTS: Surgery followed from two weeks to three years after starting occlusion therapy. Before therapy, the best visual acuity was 0.3 and the worst 0.06 (median 0.16). A median of four years after starting therapy, the visual acuity ranged from 0.9 to 0.4 (median 0.6). CONCLUSION: All milder cases profited from the treatment despite its late start. An earlier discovery would improve the prognosis. In any case, however, ophthalmologists should be encouraged to try patching before resorting to surgery. PMID- 16361186 TI - Single-stage adjustable strabismus surgery under topical anesthesia and propofol. AB - AIM: To present our experience with single-stage adjustable strabismus surgery (SSASS) under topical anesthesia and propofol. MATERIALS AND METHODS: Sixteen patients who either had diplopia before the surgery or were at risk of developing diplopia after the surgery were selected for this operation after evaluating their tolerance for an eyelid speculum. Recession of the lateral recti, recession, resection and advancement of the medial recti, anteriorization of the inferior oblique and marginal myotomy to the superior rectus were the procedures done under topical anesthesia and propofol. The operations were done in two phases; in the first phase, the patients were sedated for 10 minutes with intravenous propofol (2 mg/kg) and the operation was done under topical anesthesia with 2% lidocaine. In the second phase, the patients were conscious and the adjustments were made. RESULTS: None of the patients complained of significant pain during the surgery. No complications occurred during and after the procedure and no patients had diplopia during the postoperative follow-up. CONCLUSIONS: Single-stage adjustable surgery is practical and avoids the risks of regional and general anesthesia. The only disadvantage is the pain that some patients experience. We found that it is feasible to use propofol for this surgery; it provides deep sedation and prevents pain from being felt during the operation, and its short elimination half-life provides rapid awakening for the adjustment phase. PMID- 16361187 TI - The relationship between convergence insufficiency and ADHD. AB - BACKGROUND: Children being evaluated for attention deficit hyperactivity disorder (ADHD) often have an eye exam as part of their evaluation. The symptoms of convergence insufficiency (CI) can make it difficult for a student to concentrate on extended reading and overlap with those of ADHD. METHODS: A retrospective review of 266 patients with CI presenting to an academic pediatric ophthalmology practice was performed. All patients included were diagnosed with CI by one author (DBG) and evaluated for the diagnosis of ADHD. A computerized review was also performed looking at the converse incidence of CI in patients carrying the diagnosis of ADHD. RESULTS: We reviewed 266 charts of patients with CI. Twenty six patients (9.8%) were diagnosed with ADHD at some time in their clinical course. Of the patients with ADHD and CI, 20 (76.9%) were on medication for ADHD at the time of diagnosis for CI while 6 (23.1%) were either not on medication or the medication was discontinued several months before the diagnosis of CI. The review of computer records showed a 15.9% incidence of CI in the ADHD population. CONCLUSION: We report an apparent three-fold greater incidence of ADHD among patients with CI when compared with the incidence of ADHD in the general US population (1.8-3.3%). We also note a seeming three-fold greater incidence of CI in the ADHD population. This may simply represent an association and not be a causative relationship. Until further studies are performed, however, patients diagnosed with ADHD should be evaluated to identify the small subset that may have CI -- a condition that responds well to treatment at home. PMID- 16361188 TI - Final report of the early vs. late infantile strabismus surgery study (ELISSS), a controlled, prospective, multicenter study. AB - BACKGROUND: The optimal age for surgery for infantile esotropia is controversial. Proponents of early surgery believe that further loss of binocular vision can be prevented by early surgery, a minority believes that binocular vision can even be restored by early surgery. The ELISSS compared early with late surgery in a prospective, controlled, non-randomized, multicenter trial. METHODS: Fifty-eight clinics recruited children aged 6-18 months for the study. Each clinic operated all eligible children either 'early', i.e. at age 6-24 months, or 'late', i.e. at age 32-60 months. At baseline the angle of strabismus, refraction, degree of amblyopia and limitation of abduction were assessed. Intermediate examinations took place every six months. Children were evaluated at age six in the presence of independent observers. Primary endpoints were (i) level of binocular vision, (ii) manifest angle of strabismus at distance and (iii) remaining amblyopia. Secondary endpoints were number of operations, vertical strabismus, angle at near and the influence of surgical technique. RESULTS: A total of 231 children were recruited for early and 301 for late surgery. Age at entry examination was 11.1 months (SD 3.7 months) in the early group and 10.9 (SD 3.7) months in the late group. Refraction, amblyopia and limitation of abduction were distributed equally in the early and late groups, but the angle of strabismus was slightly larger in the early group. Dropout-rates were 26.0% in the early and 22.3% in the late group. At age six, 13.5% of the early vs. 3.9% of the late group recognized the Titmus Housefly; 3.0% of the early and 3.9% of the late group had stereopsis beyond Titmus Housefly. No significant difference was found for angle of strabismus. 35.1% of the early group and 34.8% of the late group did not have an angle between 0 degrees and 10 degrees , the thresholds set for re-operation. For ratio of the visual acuities (remaining amblyopia) there was a small but significant advantage for the early group. There was hardly any correlation between the baseline parameters and the primary endpoints. Children scheduled for early surgery had first been operated at 20 (SD 8.4) months, but 8.2% had not been operated at age six. Children scheduled for late surgery had been operated at 49.1 (SD 12.7) months, but 20.1% had not been operated at age six. The number of operations per child was 1.18 (SD 0.67) in the early and 0.99 (SD 0.64) in the late group. Age at recruitment, age that strabismus reportedly had started and refraction at entry examination were similar among operated and non-operated children. Only the angle of strabismus at entry predicted, to some extent, whether a child had been operated at age six. DISCUSSION: Children operated early had better gross stereopsis at age six as compared to children operated late. They had been operated more frequently, however, and a substantial number of children in both groups had not been operated at all. PMID- 16361189 TI - Dissertation on the cause of strabismus or squint. Translated from: "Dissertation sur la cause du strabismus ou des yeux louches". Mem. Acad. Roy. Sci. (Paris), 19 June 1743. PMID- 16361190 TI - Preparation and in vitro evaluation of thiolated chitosan microparticles. AB - The objective of this study was to prepare a microparticulate drug delivery system being based on a new thiomer, namely a chitosan 2-iminothiolane conjugate (chitosan-TBA conjugate). Due to thiol groups being immobilized on chitosan, chitosan-TBA conjugate exhibits improved mucoadhesive and permeation enhancing properties. Because of these features microparticulate drug delivery systems based on chitosan-TBA conjugate might be a promising tool for the non-invasive administration of hydrophilic macromolecular drugs. Chitosan-TBA conjugate microspheres were prepared by the emulsification/solvent evaporation method. Fluorescein-isothiocyanate labelled dextran (FITC-dextran) was chosen as a model hydrophilic drug. Microspheres have been characterized by morphological analysis, thiol group content, swelling behaviour, polymer degradation drug load determination, dissolution test and mucoadhesion studies. Results reported in this work demonstrated the possibility to obtain stable microspheres without cross-linking agents. Thiolated chitosan microspheres seem to be more stable in aqueous media with respect to unmodified chitosan. The degradability by lysozyme appears quite similar for both polymers, showing that chemical modification does not influence the biodegradable properties of chitosan. Microspheres were able to control the drug release for at least 1 h, exhibiting comparatively strong mucoadhesive properties. The chitosan-TBA conjugate microparticles remain on the mucosa in a 2.5-fold higher concentration with respect to unmodified chitosan microparticles. These data suggest that chitosan-TBA conjugate microspheres have the potential to be used as a mucoadhesive drug delivery system. PMID- 16361191 TI - Polyamide microcapsules containing jojoba oil prepared by inter-facial polymerization. AB - Jojoba oil containing polyamide microcapsules having diameter of approximately 5 microm were prepared by inter-facial polycondensation by direct method (oil-in water). Qualitative effects of both the formulation and the process parameters on microcapsules characteristics were investigated by SEM observations. Morphological analysis showed the dependence of the external membrane compactness on the chemical nature of the water-soluble polyamine and the oil-soluble acid polychloride: 1,6-hexamethylenediamine (HMDA) and terephthaloyl dichloride (TDC) were found to favour the production of smooth and dense surfaces. The use of ultrasonic irradiations during the dispersion step to get a further reduction of microcapsules size was also evaluated. PMID- 16361192 TI - Tabletted microspheres containing Cynara scolymus (var. Spinoso sardo) extract for the preparation of controlled release nutraceutical matrices. AB - Controlled release dosage forms based on tabletted microspheres containing fresh artichoke Cynara scolymus extract were performed for the oral administration of a nutritional supplement. Microspheres were prepared using a spray-drying technique; lactose or hypromellose have been chosen as excipients. Microspheres were characterized in terms of encapsulated extract content, size and morphology. Qualitative and quantitative composition of the extract before and after the spray process was determined. Compressed matrices (tablets) were prepared by direct compression of the spray-dried microspheres. In vitro release tests of microparticles and tablets prepared were carried out in both acidic and neutral media. Spray-drying is a good method to prepare microspheres containing the artichoke extract. The microspheres encapsulate an amount of extract close to the theoretical value. Particle size analyses indicate that the microparticles have dvs of approximately 6-7 microm. Electronic microscopy observations reveal that particles based on lactose have spherical shape and particles containing hypromellose are almost collapsed. The hydroalcoholic extract is stable to the microsphere production process: its polyphenolic composition (qualitative and quantitative) did not change after spraying. In vitro release studies show that microparticles characterized by a quick polyphenolic release both in acidic and neutral media due to the high water solubility of the carrier lactose. On the contrary, microspheres based hypromellose release only 20% of the loaded extract at pH 1.2 in 2 h and the total amount of polyphenols is released only after about further 6 h at pH 6.8. Matrices prepared tabletting lactose microspheres and hypromellose microparticles in the weight ratio 1:1 show a slow release rate, that lasts approximately 24 h. This one-a-day sustained release formulation containing Cynara scolymus extract could be proposed as a nutraceutical controlled release dosage form for oral administration. PMID- 16361193 TI - SLN and NLC for topical delivery of ketoconazole. AB - The clinical use of ketoconazole has been related to some adverse effects in healthy adults, specially local reactions, such as severe irritation, pruritus and stinging. The purpose of the present work is the assessment of ketoconazole stability in aqueous SLN and NLC dispersions, as well as the physicochemical stability of these lipid nanoparticles, which might be useful for targeting this drug into topical route, minimizing the adverse side effects and providing a controlled release. Lipid particles were prepared using Compritol 888 ATO as solid lipid. The natural antioxidant alpha-tocopherol was selected as liquid lipid compound for the preparation of NLC. Ketoconazole loading capacity was identical for both SLN and NLC systems (5% of particle mass). SLN were physically stable as suspensions during 3 months of storage, but the SLN matrix was not able to protect the chemically labile ketoconazole against degradation under light exposure. In contrast, the NLC were able to stabilize the drug, but the aqueous NLC dispersion showed size increase during storage. Potential topical formulations are light-protected packaged SLN or NLC physically stabilized in a gel formulation. PMID- 16361194 TI - Chitosan microspheres containing the natural urucum pigment. AB - An increasing trend in the food and pharmaceutical industries is toward replacing synthetic additives with natural products. However, in this regard, difficulties may be encountered due to the instability of such compounds. Encapsulation has become an important process to protect natural pigments. This paper reports on the encapsulation of the natural urucum pigment with chitosan using different techniques and its release under different pH conditions. The material loaded with pigment was evaluated by infrared spectroscopy, scanning electron microscopy and thermal analysis. Chitosan was found to be an effective encapsulating agent for urucum pigment. No investigations have previously been reported on the relation of chitosan to the stability of encapsulated natural pigments. PMID- 16361195 TI - Formulation and evaluation of sustained release microspheres of poly-lactide-co glycolide containing tamoxifen citrate. AB - Tamoxifen citrate, a non-steroidal anti-oestrogen has potential applications in treatment of breast cancer. Biodegradable microspheres of' PLGA 65:35 were prepared by o/w emulsification solvent evaporation method. In this study, different batches of varying concentration of drug, polymer, polyvinyl alcohol and solvent were prepared. All the batches prepared were characterized by particle size distribution, encapsulation efficiency and in vitro release behaviour. Drug, polymer and PVA concentrations were varied to obtain optimum release profile for sustaining the action of drug. PMID- 16361196 TI - Preparation and characterization of ibuprofen microspheres. AB - Ibuprofen was microencapsulated with Eudragit RS using an o/w emulsion solvent evaporation technique. The effects of three formulation variables including the drug:polymer ratio, emulsifier (polyvinyl alcohol) concentration and organic solvent (chloroform) volume on the entrapment efficiency and microspheres size distribution were examined. The drug release rate from prepared microspheres and the release kinetics were also studied. The results demonstrated that microspheres with good range of particle size can be prepared, depending on the formulation components. The drug:polymer ratio had a considerable effect on the entrapment efficiency. However, particle size distribution of microspheres was more dependent on the volume of chloroform and polyvinyl alcohol concentration rather than the drug:polymer ratio. The drug release pattern showed a burst effect for all prepared microspheres due to the presence of uncovered drug crystals on the surface. It was shown that the release profiles of all formulations showed good correlation with the Higuchi model of release. PMID- 16361197 TI - In-vitro analysis of APA microcapsules for oral delivery of live bacterial cells. AB - Oral administration of microcapsules containing live bacterial cells has potential as an alternative therapy for several diseases. This article evaluates the suitability of the alginate-poly-L-lysine-alginate (APA) microcapsules for oral delivery of live bacterial cells, in-vitro, using a dynamic simulated human gastro-intestinal (GI) model. Results showed that the APA microcapsules were morphologically stable in the simulated stomach conditions, but did not retain their structural integrity after a 3-day exposure in simulated human GI media. The microbial populations of the tested bacterial cells and the activities of the tested enzymes in the simulated human GI suspension were not substantially altered by the presence of the APA microcapsules, suggesting that there were no significant adverse effects of oral administration of the APA microcapsules on the flora of the human gastrointestinal tract. When the APA microcapsules containing Lactobacillus plantarum 80 (LP80) were challenged in the simulated gastric medium (pH = 2.0), 80.0% of the encapsulated cells remained viable after a 5-min incubation; however, the viability decreased considerably (8.3%) after 15 min and dropped to 2.6% after 30 min and lower than 0.2% after 60 min, indicating the limitations of the currently obtainable APA membrane for oral delivery of live bacteria. Further in-vivo studies are required before conclusions can be made concerning the inadequacy of APA microcapsules for oral delivery of live bacterial cells. PMID- 16361198 TI - Spray-dried chitosan/ethylcellulose microspheres for nasal drug delivery: swelling study and evaluation of in vitro drug release properties. AB - The aim of this study was to develop spray-dried chitosan-based microspheres, suitable for nasal delivery of loratadine, and to evaluate their potential of modifying loratadine release. The microspheres were composed with ethylcellulose (EC) and chitosan (CM) in two different weight ratios, 1:2 and 1:3. One-phase systems (dispersions) and two-phase systems (emulsions and suspensions) were subjected to spray-drying, resulting in conventional and composed microspheres, respectively. The microspheres were evaluated with respect to the yield, particle size, encapsulation efficiency, physical state of the drug in the polymer matrix, swelling properties and in vitro drug release profile. It was shown that particle size, swelling ability and loratadine release from spray-dried microspheres were significantly affected by the polymeric composition and feed concentration in spray-drying process. Emulsifying method to produce composed EC/CM microspheres resulted in improved loratadine entrapment and moderate swelling, when compared to conventional chitosan microspheres. It seems like better formation of EC cores and chitosan coating were obtained when higher feed concentration and ultrasonic homogenization were employed in the preparation of emulsion systems and when EC to CM weight ratio was 1:3. PMID- 16361202 TI - Natural medicines for alcoholism treatment. PMID- 16361204 TI - Cannabis use in adolescents: the impact of risk and protective factors and social functioning. AB - The study uses a school-based sample to test the social and familial risk and protective factors relating to cannabis use. Based on a self-completion survey of 2078 14-16-year-olds (mean age of 15 years) attending seven standard state-run secondary schools in south London, an assessment was made of rates and risk factors for cannabis use. Twenty-four per cent of the total sample had ever used cannabis, with 15% having done so in the month prior to assessment. In addition to greater likelihood of illicit drug use, lifetime cannabis users were less likely to spend time regularly with both their mothers and fathers, but more likely to spend free time with friends who smoked, drank alcohol and used illicit drugs, and with friends involved in criminal activities. Among those who had ever used cannabis, frequency of cannabis use was predicted (using linear regression) by two onset factors (earlier initiation of drinking and cannabis use were both linked to more frequent use) and two social factors (more time spent with drug using friends and less time spent with the mother). Overall, the study showed that early onset, itself predicted by social networks, is linked to more frequent use of cannabis and that this appears to be sustained by less time spent with parents and more with drug-using peers. PMID- 16361203 TI - Process evaluation of an out-patient detoxification service. AB - This paper describes the process evaluation of an out-patient detoxification service (ODS) established by Drug Health Services (DHS) to increase the supervised withdrawal options for substance users in a Sydney metropolitan Area Health Service. The ODS aimed to provide a safe and effective supervised withdrawal to substance users who were at low risk of severe withdrawal, engage those with severe dependence in further treatment and increase the involvement of general practitioners (GPs) in the medical care of ODS clients. During its first 10 months of operation, the ODS received 199 inquiries, assessed 82 individuals and admitted 76 clients for detoxification. Withdrawal treatment proceeded without complications and within the expected time frames. Fifty-four clients completed withdrawal, 10 ceased treatment, 10 remained in treatment without completing withdrawal and two were transferred elsewhere. Clients who injected substances (mainly heroin) daily at admission, compared to others, were less likely to complete withdrawal and more likely to use a range of non-prescribed substances during withdrawal. One-fifth of clients went on to further treatment with DHS, attending at least once. Overall, the ODS met its goals, providing a safe and effective supervised withdrawal to local residents, especially women, young people and those withdrawing from benzodiazepines who had significant substance dependence, impairment and previous alcohol and other drug (AOD) treatment. Non-injecting substance users benefited most from the ODS in terms of withdrawal completion and ongoing treatment. The level of GP involvement in the conjoint care of ODS clients remained constant over time. The development and expansion of the ODS are discussed. PMID- 16361205 TI - Alcohol-related associative strength and drinking behaviours: concurrent and prospective relationships. AB - The first part of this research assessed the longitudinal relationships between alcohol-related associative strength and alcohol use measured at two time-points, 6 months apart. Cross-lagged results support the utility of alcohol-related associative strength to predict drinking behaviours prospectively and vice versa. These results remained after competing explanations of previous use, autocorrelations between memory measures, sensation seeking and background variables of age and gender were accounted for. Findings offer further evidence for an implicit cognitions approach to drinking processes. In the second part of our study, cross-sectional analysis investigated potential mediating mechanisms in the relation of associative strength to quantity and frequency dimensions of drinking. Mediational models provide preliminary evidence that implicit memory processes may have differential effects on quantity and frequency dimensions of drinking behaviours. The results point to the possibility that increasing awareness of implicit alcohol-related associations may have utility in interventions for young adults. PMID- 16361206 TI - The management of alcohol, tobacco and illicit drug use problems by general practitioners in Australia. AB - The aim of this study was to document the frequency of the management of illicit drug, alcohol and tobacco problems in general practice in Australia. Data from the Bettering the Evaluation and Care of Health (BEACH) study of general practice, April 1998 to March 2003, were analysed. BEACH is an ongoing national study of general practice in Australia. Each year a random sample of approximately 1000 general practitioners (GPs) participate, each providing details of 100 patient encounters. Samples are drawn from the Medicare data held by the Health Insurance Commission. Patient demographic breakdowns, medication, other treatment, referrals and other medical procedures ordered were examined for all problems labelled by GPs as illicit, alcohol and tobacco problems. Annually in Australia, it was estimated that 615,000 GP encounters--or 0.6% of all encounters--involved the management of illicit drug use problems presumably most commonly for problematic heroin use. Despite a much higher population prevalence of use and use disorders, the management of alcohol or tobacco use problems was less common, with 0.4% and 0.3% of encounters, respectively, comprising treatment of these problems. Clear demographic differences existed across the groups. The management of problems also differed, with illicit drug use problems more likely to involve provision of medication, and alcohol and tobacco treatment more likely to involve counselling and/or health advice. Despite higher rates of alcohol and tobacco use problems among patients seeing GPs in Australia, the rate of treatment for such problems was relatively lower than it was for illicit drug use problems. More efforts need to be directed towards assisting GPs to identify and target problematic alcohol and tobacco use among their patients. PMID- 16361207 TI - Drug 'hot-spots', alcohol availability and violence. AB - Ecological studies have shown a relationship between alcohol outlet densities and violence and between the location of crimes related to illicit drug use (so called 'hot spots') and violence. To date, no study has compared the effects of alcohol outlets and drug hot spots on rates of violence. The present study examined this relationship in the City of Houston, Texas. An ecological study design was employed, using a sample of 439 census tracts from Houston, Texas. Neighborhood socio-structural, alcohol outlet density, drug crime density and violent crime density data were collected from archival sources and analyzed using multivariate and spatial statistics. Using ordinary least-squares analysis, the neighborhood socio-structural covariates explained about 40% of the variability in violent crime. Adding alcohol outlet density in the target census tracts explained an additional 6%, while the addition of drug crime density explained an additional 32%. In the final model, that controlled for the effects of autocorrelated error, both drug crime density in the target and adjacent census tracts remained significant predictors of violent crime, while only off sale density in the target census tract remained significant in the model. The findings indicate that drug crime density explained a greater amount of variance in violent crime rates than the alcohol outlet density. The methodological and policy implications of these findings are discussed, along with the shortcomings of the analysis presented. PMID- 16361208 TI - Youth alcohol and other drug use in the United States and Australia: a cross national comparison of three state-wide samples. AB - Although youth drug and alcohol harm minimization policies in Australia are often contrasted with the abstinence and zero tolerance policies adopted in the United States, there has been little research directly comparing youth substance use behaviour in the two countries. Three state representative samples in Victoria, Australia (n = 7898) and in the US states of Oregon (n = 15,224) and Maine (n = 16,245) completed a common cross-sectional student survey. Rates of alcohol use (lifetime alcohol use, recent use in the past 30 days), alcohol use exceeding recommended consumption limits (binge drinking: five or more drinks in a session), other licit drug use (tobacco use), and norm-violating substance use (substance use at school, use in the past 30 days of marijuana or other illicit drug use) were compared for males and females at ages 12-17. Rates were lower (odds ratios 0.5-0.8) for youth in Maine and Oregon compared to Victoria for lifetime and recent alcohol use, binge drinking and daily cigarette smoking. However, rates of recent marijuana use and recent use of other illicit drugs were higher in Maine and Oregon, as were reports of being drunk or high at school. In contradiction of harm minimization objectives, Victoria, relative to the US states of Oregon and Maine, demonstrated higher rates of alcohol use exceeding recommended consumption limits and daily tobacco use. However, findings suggested that aspects of norm-violating substance use (substance use at school, marijuana use and other illicit drug use) were higher in the US states compared to Victoria. PMID- 16361209 TI - Natural medicines for alcoholism treatment: a review. AB - Alcoholism is a serious problem throughout the world. The development of alcoholism remedies have medical, social and economical significance. In view of the pitfalls of psychological dependence and adverse behavioural effects of synthetic drugs, the development of low toxicity and high efficiency medicines derived from natural products exhibits expansive market prospects. Based on these considerations, we summarize briefly folk application of traditional hangover remedies and clinical application of herbal complex and patent medicines for alcoholism treatment. We have reviewed the effects of natural medicines on intake, absorption and metabolism of alcohol, as well as the protective effects on alcohol-induced acute and chronic tissue injury. PMID- 16361210 TI - Mortality and population drinking: a review of the literature. AB - The aim of this review was to review research addressing the relationship between population drinking and health, particularly mortality. The review is based primarily on articles published in international journals after 1994 to February 2005, identified via Medline. The method used in most studies is time-series analysis based on autoregressive intergrated moving average (ARIMA) modelling. The outcome measures covered included the following mortality indicators: mortality from liver cirrhosis and other alcohol-related diseases, accident mortality, suicide, homicide, ischaemic heart disease (IHD) mortality and all cause mortality. The study countries included most of the EU member states as of 1995 (14 countries), Canada and the United States. For Eastern Europe there was only scanty evidence. The study period was in most cases the post-war period. There was a statistically significant relationship between per capita consumption and mortality from liver cirrhosis and other alcohol-related diseases in all countries. In about half the countries, there was a significant relationship between consumption, on one hand, and mortality from accidents and homicide as well as all-cause mortality on the other hand. A link between alcohol and suicide was found in all regions except for mid- and southern Europe. There was no systematic link between consumption and IHD mortality. Overall, a 1-litre increase in per capita consumption was associated with a stronger effect in northern Europe and Canada than in mid- and southern Europe. Research during the past decade has strengthened the notion of a relationship between population drinking and alcohol-related harm. At the same time, the marked regional variation in the magnitude of this relationship suggests the importance of drinking patterns for modifying the impact of alcohol. By and large, there was little evidence for any cardioprotective effect at the population level. It is a challenge for future research to reconcile this outcome with the findings from observational studies, most of which suggest a protective effect of moderate drinking. PMID- 16361211 TI - Ushering in another harm reduction era? Discursive authenticity, drug policy and research. PMID- 16361212 TI - Moral frameworks, ethical engagement and harm reduction: commentary on 'Ethical challenges and responses in harm reduction research: promoting applied communitarian ethics' by C. L. Fry, C. Treloar & L. Maher. PMID- 16361213 TI - Harm reduction ethics: a promising basis for drug policy. PMID- 16361214 TI - Implementation of brief alcohol intervention in primary health care: do nurses' and general practitioners' attitudes, skills and knowledge change? AB - Brief alcohol intervention reduces heavy drinking, but its implementation has been challenging. The purpose was to evaluate self-reported changes in attitudes, skills and knowledge regarding brief intervention among nurses and general practitioners (GPs) during an implementation project. A questionnaire survey was used before and after the implementation to all nurses and GPs working at the time in the seven primary health-care centres of the city of Tampere, Finland. Several positive changes indicate an increased amount of knowledge regarding brief intervention among the professionals during the implementation. This was found especially among the nurses. The success in increasing the knowledge can also be seen in a decrease of training needs. Instead, attitudes and skills among the professionals did not seem to develop positively. Increasing motivational skills especially seems to be the future challenge. PMID- 16361215 TI - Alcohol policy and harm reduction in Australia. AB - With consultations having been held across Australia this year as part of the process of developing a new National Alcohol Strategy, it seemed timely to invite my colleagues from the National Drug Research Institute who are experts in the alcohol field to write this Harm Reduction Digest. The authors have canvassed a range of alcohol policy options and discussed their effectiveness in reducing harm for what is arguably Australia's number one drug problem. Australia's response to alcohol and other drug problems has, historically, been based on 'harm minimization--incorporating supply reduction, demand reduction and harm reduction'. At this time where the policy options for alcohol are being set for the next 5 years in a climate of 'small government', removing restrictions of 'fair competition' in business and a belief in the free market, what does the research have to say about recommended policies and strategies to reduce alcohol related harm? PMID- 16361217 TI - Treatment options for metastatic tumors to the choroid. AB - Metastatic tumors are generally considered to be the most common intraocular tumors. These tumors may metastasize from a variety of different primary sites with breast carcinoma being the most common primary malignancy in women and lung cancer being the most common primary malignancy in men. Treatment options for these patients include systemic therapy, conventional external beam radiation, plaque brachytherapy, proton beam irradiation, photodynamic therapy, transpupillary thermotherapy, and laser photocoagulation. Overall, treatment of patients with metastatic tumors to the choroid has proven to be effective in both improving the patient's visual acuity and quality of life. PMID- 16361218 TI - An evaluation of analytical methods, air sampling techniques, and airborne occupational exposure of metalworking fluids. AB - This article summarizes an assessment of air sampling and analytical methods for both oil and water-based metalworking fluids (MWFs). Three hundred and seventy four long-term area and personal airborne samples were collected at four plants using total (closed-face) aerosol samplers and thoracic samplers. A direct reading device (DustTrak) was also used. The processes sampled include steel tube making, automotive component manufacturing, and small part manufacturing in a machine shop. The American Society for Testing and Materials (ASTM) Method PS42 97 of analysis was evaluated in the laboratory. This evaluation included sample recovery, determination of detection limits, and stability of samples during storage. Results of the laboratory validation showed (a) the sample recovery to be about 87%, (b) the detection limit to be 35 microg, and (c) sample stability during storage at room temperature to decline rapidly within a few days. To minimize sample loss, the samples should be stored in a freezer and analyzed within a week. The ASTM method should be the preferred method for assessing metalworking fluids (MWFs). The ratio of thoracic aerosol to total aerosol ranged from 0.6 to 0.7. A similar relationship was found between the thoracic extractable aerosol and total extractable aerosol. The DustTrak, with 10-microm sampling head, was useful in pinpointing the areas of potential exposure. MWF exposure at the four plants ranged from 0.04 to 3.84 mg/m3 with the geometric mean ranging between 0.22 to 0.59 mg/m3. Based on this data and the assumption of log normality, MWF exposures are expected to exceed the National Institute for Occupational Safety and Health recommended exposure limit of 0.5 mg/m3 as total mass and 0.4 mg/m3 as thoracic mass about 38% of the time. In addition to controlling airborne MWF exposure, full protection of workers would require the institution of programs for fluid management and dermal exposure prevention. PMID- 16361219 TI - Blood lead levels in NASCAR Nextel Cup teams. AB - This pilot study determines whether NASCAR racing teams demonstrate exposure to lead from exhaust by evaluation of blood lead levels (BLL). Participants were stratified by proximity to fuel exhaust or whether they worked on an engine. Each participant completed a self-reported survey recording demographics, lead exposure (occupational or in-home environment), and any physical symptoms of lead toxicity. Blood lead levels were then measured. BLL of 47 individuals ranged from 1-22 microg/dL with a median of 9.4 microg/dL. Nineteen of 47 (40.4%) had BLL > or = 10 microg/dL. Participants exposed to exhaust gas had the highest relative risks (RR) for elevated lead, followed by working on brakes and radiator repair. The RR of having an elevated BLL and self-reported adverse health outcomes or symptoms was increased. This study of NASCAR racing teams demonstrates lead exposure. PMID- 16361220 TI - Upper limb dynamic responses to impulsive forces for selected assembly workers. AB - This study evaluated the upper limb, dynamic, mechanical response parameters for 14 male assembly workers recruited from selected jobs based on power tool use. It was hypothesized that the type of power tool operation would affect stiffness, effective mass, and damping of the upper extremity; and workers with symptoms and positive physical examination findings would have different mechanical responses than asymptomatic workers without physical examination findings. Participants included operators who regularly used torque reaction power hand tools, such as nutrunners and screwdrivers, and nontorque reaction power hand tools, such as riveters. The mechanical parameters of the upper limb were characterized from the loading response of an apparatus having known dynamic properties while worker grasps an oscillating handle in free vibration. In addition, all workers underwent a physical examination, magnetic resonance imaging, and completed a symptom survey. Workers were categorized as controls or cases based on reported forearm symptoms and physical exam findings. A total of seven workers were categorized as cases and had less average mechanical stiffness (46%, p > 0.01), damping (74%, p > 0.01), and effective mass (59%, p > 0.05) than the seven workers categorized as controls. Magnetic resonance imaging (MRI) findings suggestive of muscle edema were observed for two workers classified as cases and who regularly used torque reaction power tools. No MRI enhancement was observed in the seven subjects who did not regularly use torque reaction power tools. The ergonomic consequences of less stiffness, effective mass, and damping in symptomatic workers may include reduced capacity to react against rapidly building torque reaction forces encountered when operating power hand tools. PMID- 16361221 TI - Relationship of surgical approach to neurodevelopmental outcomes in hypoplastic left heart syndrome. AB - OBJECTIVE: Two strategies for surgical management are used for infants with hypoplastic left heart syndrome (HLHS), primary heart transplantation and the Norwood procedure. We sought to determine how these 2 surgical approaches influence neurodevelopmental outcomes at school age. METHODS: A multicenter, cross-sectional study of neurodevelopmental outcomes among school-aged children (>8 years of age) with HLHS was undertaken between July 2003 and September 2004. Four centers enrolled 48 subjects, of whom 47 completed neuropsychologic testing. Twenty-six subjects (55%) had undergone the Norwood procedure and 21 (45%) had undergone transplantation, with an intention-to-treat analysis. The mean age at testing was 12.4 +/- 2.5 years. Evaluations included the Wechsler Abbreviated Scale of Intelligence, Clinical Evaluation of Language Fundamentals, Wechsler Individual Achievement Test, and Beery-Buktenica Developmental Test of Visual Motor Integration. RESULTS: The mean neurocognitive test results were significantly below population normative values. The mean full-scale IQ for the entire cohort was 86 +/- 14. In a multivariate model, there was no association of surgical strategy with any measure of developmental outcome. A longer hospital stay, however, was associated significantly with lower verbal, performance, and full-scale IQ scores. Aortic valve atresia was associated with lower math achievement test scores. CONCLUSIONS: Neurodevelopmental deficits are prevalent among school-aged children with HLHS, regardless of surgical approach. Complications that result in prolonged hospitalization at the time of the initial operation are associated with neurodevelopmental status at school age. PMID- 16361222 TI - Areca-nut abuse and neonatal withdrawal syndrome. AB - Areca-nut chewing occurs widely in South Asia and the Indian subcontinent. Here we present a case of neonatal withdrawal syndrome in an infant born to a woman who was a chronic areca-nut user. Arecoline, the principal neuroactive alkaloid in areca nuts, was found in the mother's placenta. PMID- 16361223 TI - Safety and tolerability of 1% pimecrolimus cream among infants: experience with 1133 patients treated for up to 2 years. AB - Pimecrolimus is a calcineurin inhibitor developed for the topical treatment of atopic dermatitis. During the clinical development of 1% pimecrolimus cream, 1133 patients 3 to 23 months of age with mild to severe atopic dermatitis were treated for up to 2 years. The objective of this review is to discuss the safety and tolerability of 1% pimecrolimus cream among infants, on the basis of the combined results from all studies (4 pharmacokinetic studies and 6 clinical trials) conducted among these patients. Pimecrolimus blood concentrations measured for 35 patients were consistently low (< or =1 ng/mL in >80% of samples), irrespective of the disease severity and extent, and remained low during intermittent treatment for up to 1 year. The level of systemic exposure to pimecrolimus among infants was comparable to that observed for older pediatric patients enrolled in the same studies and treated in the same way with 1% pimecrolimus cream, which indicated that young pediatric patients are not at higher risk of significant percutaneous absorption of topically applied pimecrolimus, despite their large skin surface area/body mass ratio. The 6 clinical trials included a total of 1098 infants, who were treated for periods ranging from 4 weeks to 2 years. Most of these patients (60%) had moderate to severe disease at baseline. The most frequently reported adverse events were common childhood disorders such as nasopharyngitis, pyrexia, upper respiratory tract infections, ear infections, and bronchitis. During the double-blind (DB) studies or DB phases of studies, the incidence rates for the most frequently reported adverse events were similar for patients who received 1% pimecrolimus cream and patients who received the vehicle, except for the incidence of teething, which was higher among the pimecrolimus-treated infants (relative risk: 2.02; 95% confidence interval: 1.32 3.27). Treatment with 1% pimecrolimus cream was not associated with an increase in the overall incidence of nonskin infections, compared with the vehicle (relative risk: 1.015; 95% confidence interval: 0.88-1.18). The incidence density (ID) rates for total bacterial, fungal, parasitic, and viral skin infections during the DB studies or DB phases of studies were comparable for patients treated with 1% pimecrolimus cream and patients who received the vehicle. The ID rate of herpes simplex virus infections was 0.8 cases per 1000 patient-months of follow-up monitoring among patients treated with 1% pimecrolimus cream and 1.7 cases per 1000 patient-months of follow-up monitoring among patients who received the vehicle. Considering all 1098 infants treated with 1% pimecrolimus cream in DB trials and open-label studies, the ID rate of clinically diagnosed eczema herpeticum was 1.3 cases per 1000 patient-months of follow-up monitoring. Burning and erythema were the most frequently reported application site reactions, with ID rates of 2.0 and 1.2 cases per 1000 patient-months of follow-up monitoring, respectively. No sign of immunosuppression was found among infants treated intermittently with 1% pimecrolimus cream for up to 2 years; they demonstrated normal immune responses to vaccinations and did not show increases in the incidence of systemic infections or skin infections over time. PMID- 16361224 TI - Increased prevalence of fetal thrombi in monochorionic-twin placentas. AB - OBJECTIVE: Fetal vessel thrombosis has been related to pregnancy complications and adverse neonatal outcomes in singleton pregnancies, but the clinical and pathologic characteristics of fetal thrombosis in twin pregnancy are still ill defined. METHODS: To clarify the characteristics of fetal thrombosis in twins, we reviewed the histology slides and medical records of live-born infants of twin pregnancies involving the live birth of at least 1 fetus (monochorionic: 389 cases; dichorionic: 780 cases) and singleton pregnancies (1162 cases). RESULTS: The incidence of fetal thrombosis in monochorionic-twin pregnancies was significantly higher than that of dichorionic-twin and singleton pregnancies (5.1% vs 3.0% and 2.8%, respectively). The incidence of velamentous umbilical cord insertion in monochorionic- and dichorionic-twin placentas was also higher than in singleton placentas (12% and 7% vs 2%, respectively). Fetal thrombosis in twin placentas was associated with intrauterine growth restriction. In monochorionic twins, fetal thrombosis was associated with co-twin fetal death, but in dichorionic twins no correlation was identified. Microscopically, fetal vessel thrombosis in twin placentas was associated with vascular cushions (fibrous hyperplasia of fetal vessel), as is the case occasionally in singletons. CONCLUSIONS: The incidence of fetal vessel thrombosis in monochorionic placentas was higher than that seen in dichorionic-twin placentas and singleton gestations. Fetal vessel thrombosis in twin pregnancies showed a correlation with intrauterine growth restriction, peripheral cord insertions, and major vascular cushions. PMID- 16361225 TI - These are the technologies that try men's souls: common-sense health information technology. PMID- 16361226 TI - Genomewide screen reveals a wide regulatory network for di/tripeptide utilization in Saccharomyces cerevisiae. AB - Small peptides of two to six residues serve as important sources of amino acids and nitrogen required for growth by a variety of organisms. In the yeast Saccharomyces cerevisiae, the membrane transport protein Ptr2p, encoded by PTR2, mediates the uptake of di/tripeptides. To identify genes involved in regulation of dipeptide utilization, we performed a systematic, functional examination of this process in a haploid, nonessential, single-gene deletion mutant library. We have identified 103 candidate genes: 57 genes whose deletion decreased dipeptide utilization and 46 genes whose deletion enhanced dipeptide utilization. On the basis of Ptr2p-GFP expression studies, together with PTR2 expression analysis and dipeptide uptake assays, 42 genes were ascribed to the regulation of PTR2 expression, 37 genes were involved in Ptr2p localization, and 24 genes did not apparently affect Ptr2p-GFP expression or localization. The 103 genes regulating dipeptide utilization were distributed among most of the Gene Ontology functional categories, indicating a very wide regulatory network involved in transport and utilization of dipeptides in yeast. It is anticipated that further characterization of how these genes affect peptide utilization should add new insights into the global mechanisms of regulation of transport systems in general and peptide utilization in particular. PMID- 16361227 TI - Analysis of pyrimidine catabolism in Drosophila melanogaster using epistatic interactions with mutations of pyrimidine biosynthesis and beta-alanine metabolism. AB - The biochemical pathway for pyrimidine catabolism links the pathways for pyrimidine biosynthesis and salvage with beta-alanine metabolism, providing an array of epistatic interactions with which to analyze mutations of these pathways. Loss-of-function mutations have been identified and characterized for each of the enzymes for pyrimidine catabolism: dihydropyrimidine dehydrogenase (DPD), su(r) mutants; dihydropyrimidinase (DHP), CRMP mutants; beta-alanine synthase (betaAS), pyd3 mutants. For all three genes, mutants are viable and fertile and manifest no obvious phenotypes, aside from a variety of epistatic interactions. Mutations of all three genes disrupt suppression by the rudimentary gain-of-function mutation (r(Su(b))) of the dark cuticle phenotype of black mutants in which beta-alanine pools are diminished; these results confirm that pyrimidines are the major source of beta-alanine in cuticle pigmentation. The truncated wing phenotype of rudimentary mutants is suppressed completely by su(r) mutations and partially by CRMP mutations; however, no suppression is exhibited by pyd3 mutations. Similarly, su(r) mutants are hypersensitive to dietary 5 fluorouracil, CRMP mutants are less sensitive, and pyd3 mutants exhibit wild-type sensitivity. These results are discussed in the context of similar consequences of 5-fluoropyrimidine toxicity and pyrimidine catabolism mutations in humans. PMID- 16361228 TI - The integral membrane protein Pom34p functionally links nucleoporin subcomplexes. AB - Here we have examined the function of Pom34p, a novel membrane protein in Saccharomyces cerevisiae, localized to nuclear pore complexes (NPCs). Membrane topology analysis revealed that Pom34p is a double-pass transmembrane protein with both the amino (N) and carboxy (C) termini positioned on the cytosolic/pore face. The network of genetic interactions between POM34 and genes encoding other nucleoporins was established and showed specific links between Pom34p function and Nup170p, Nup188p, Nup59p, Gle2p, Nup159p, and Nup82p. The transmembrane domains of Pom34p in addition to either the N- or C-terminal region were necessary for its function in different double mutants. We further characterized the pom34deltaN nup188delta mutant and found it to be perturbed in both NPC structure and function. Mislocalization of a subset of nucleoporins harboring phenylalanine-glycine repeats was observed, and nuclear import capacity for the Kap104p and Kap121p pathways was inhibited. In contrast, the pom34delta pom152delta double mutant was viable at all temperatures and showed no such defects. Interestingly, POM152 overexpression suppressed the synthetic lethality of pom34delta nup170delta and pom34delta nup59delta mutants. We speculate that multiple integral membrane proteins, either within the nuclear pore domain or in the nuclear envelope, execute coordinated roles in NPC structure and function. PMID- 16361229 TI - Glc7-Reg1 phosphatase signals to Yck1,2 casein kinase 1 to regulate transport activity and glucose-induced inactivation of Saccharomyces maltose permease. AB - The Saccharomyces casein kinase 1 isoforms encoded by the essential gene pair YCK1 and YCK2 control cell growth and morphogenesis and are linked to the endocytosis of several membrane proteins. Here we define roles for the Yck1,2 kinases in Mal61p maltose permease activation and trafficking, using a yck1delta yck2-2(ts) (yck(ts)) strain with conditional Yck activity. Moreover, we provide evidence that Glc7-Reg1 phosphatase acts as an upstream activator of Yck1,2 kinases in a novel signaling pathway that modulates kinase activity in response to carbon source availability. The yck(ts) strain exhibits significantly reduced maltose transport activity despite apparently normal levels and cell surface localization of maltose permease protein. Glucose-induced internalization and rapid loss of maltose transport activity of Mal61/HAp-GFP are not observed in the yck(ts) strain and maltose permease proteolysis is blocked. We show that a reg1delta mutant exhibits a phenotype remarkably similar to that conferred by yck(ts). The reg1delta phenotype is not enhanced in the yck(ts) reg1delta double mutant and is suppressed by increased Yck1,2p dosage. Further, although Yck2p localization and abundance do not change in the reg1delta mutant, Yck1,2 kinase activity, as assayed by glucose-induced HXT1 expression and Mth1 repressor stability, is substantially reduced in the reg1delta strain. PMID- 16361230 TI - Fold recognition of the human immunodeficiency virus type 1 V3 loop and flexibility of its crown structure during the course of adaptation to a host. AB - The third hypervariable (V3) region of the HIV-1 gp120 protein is responsible for many aspects of viral infectivity. The tertiary structure of the V3 loop seems to influence the coreceptor usage of the virus, which is an important determinant of HIV pathogenesis. Hence, the information about preferred conformations of the V3 loop region and its flexibility could be a crucial tool for understanding the mechanisms of progression from an initial infection to AIDS. Taking into account the uncertainty of the loop structure, we predicted the structural flexibility, diversity, and sequence fitness to the V3-loop structure for each of the sequences serially sampled during an asymptomatic period. Structural diversity correlated with sequence diversity. The predicted crown structure usage implied that structural flexibility depended on the patient and that the antigenic character of the virus might be almost uniform in a patient whose immune system is strong. Furthermore, the predicted structural ensemble suggested that toward the end of the asymptomatic period there was a change in the V3-loop structure or in the environment surrounding the V3 loop, possibly because of its proximity to the gp120 core. PMID- 16361231 TI - Chromosome-level homeology in paleopolyploid soybean (Glycine max) revealed through integration of genetic and chromosome maps. AB - Soybean has 20 chromosome pairs that are derived from at least two rounds of genomewide duplication or polyploidy events although, cytogenetically, soybean behaves like a diploid and has disomic inheritance for most loci. Genetically anchored genomic clones were used as probes for fluorescence in situ hybridization (FISH) to determine the level of postpolyploid chromosomal rearrangements and to integrate the genetic and physical maps to (1) assign linkage groups to specific chromosomes, (2) assess chromosomal structure, and (3) determine the distribution of recombination along the length of a chromosome. FISH mapping of seven putatively gene-rich BACs from linkage group L (chromosome 19) revealed that most of the genetic map correlates to the highly euchromatic long arm and that there is extensive homeology with another chromosome pair, although colinearity of some loci does appear to be disrupted. Moreover, mapping of BACs containing high-copy sequences revealed sequestration of high-copy repeats to the pericentromeric regions of this chromosome. Taken together, these data present a model of chromosome structure in a highly duplicated but diploidized eukaryote, soybean. PMID- 16361232 TI - Pleiotropic quantitative trait loci contribute to population divergence in traits associated with life-history variation in Mimulus guttatus. AB - Evolutionary biologists seek to understand the genetic basis for multivariate phenotypic divergence. We constructed an F2 mapping population (N = 539) between two distinct populations of Mimulus guttatus. We measured 20 floral, vegetative, and life-history characters on parents and F1 and F2 hybrids in a common garden experiment. We employed multitrait composite interval mapping to determine the number, effect, and degree of pleiotropy in quantitative trait loci (QTL) affecting divergence in floral, vegetative, and life-history characters. We detected 16 QTL affecting floral traits; 7 affecting vegetative traits; and 5 affecting selected floral, vegetative, and life-history traits. Floral and vegetative traits are clearly polygenic. We detected a few major QTL, with all remaining QTL of small effect. Most detected QTL are pleiotropic, implying that the evolutionary shift between these annual and perennial populations is constrained. We also compared the genetic architecture controlling floral trait divergence both within (our intraspecific study) and between species, on the basis of a previously published analysis of M. guttatus and M. nasutus. Eleven of our 16 floral QTL map to approximately the same location in the interspecific map based on shared, collinear markers, implying that there may be a shared genetic basis for floral divergence within and among species of Mimulus. PMID- 16361233 TI - Gene action of new mutations in Arabidopsis thaliana. AB - For a newly arising mutation affecting a trait under selection, its degree of dominance relative to the preexisting allele(s) strongly influences its evolutionary impact. We have estimated dominance parameters for spontaneous mutations in a subset of lines derived from a highly inbred founder of Arabidopsis thaliana by at least 17 generations of mutation accumulation (MA). The labor-intensive nature of the crosses and the anticipated subtlety of effects limited the number of MA lines included in this study to 8. Each MA line was selfed and reciprocally crossed to plants representing the founder genotype, and progeny were assayed in the greenhouse. Significant mutational effects on reproductive fitness included a recessive fitness-enhancing effect in one line and fitness-reducing effects, one additive and the other slightly recessive. Mutations conferring earlier phenology or smaller leaves were significantly recessive. For effects increasing leaf number and reducing height at flowering, additive gene action accounted for the expression of the traits. The sole example of a significantly dominant mutational effect delayed phenology. Our findings of recessive action of a fitness-enhancing mutational effect and additive action of a deleterious effect counter a common expectation of (partial) dominance of alleles that increase fitness, but the frequency of occurrence of such mutations is unknown. PMID- 16361234 TI - New Arabidopsis recombinant inbred line populations genotyped using SNPWave and their use for mapping flowering-time quantitative trait loci. AB - The SNPWave marker system, based on SNPs between the reference accessions Colombia-0 and Landsberg erecta (Ler), was used to distinguish a set of 92 Arabidopsis accessions from various parts of the world. In addition, we used these markers to genotype three new recombinant inbred line populations for Arabidopsis, having Ler as a common parent that was crossed with the accessions Antwerp-1, Kashmir-2, and Kondara. The benefit of using multiple populations that contain many similar markers and the fact that all markers are linked to the physical map of Arabidopsis facilitates the quantitative comparison of maps. Flowering-time variation was analyzed in the three recombinant inbred line populations. Per population, four to eight quantitative trait loci (QTL) were detected. The comparison of the QTL positions related to the physical map allowed the estimate of 12 different QTL segregating for flowering time for which Ler has an allele different from one, two, or three of the other accessions. PMID- 16361235 TI - Chromosome segment duplications in Neurospora crassa and their effects on repeat induced point mutation and meiotic silencing by unpaired DNA. AB - The size and extent of four Neurospora crassa duplications, Dp(AR17), Dp(IBj5), Dp(OY329), and Dp(B362i), was determined by testing the coverage of RFLP markers. The first three duplications were all > approximately 350 kb and have been shown in earlier studies to act as dominant suppressors of repeat-induced point mutation (RIP) in gene-sized duplications, possibly via titration of the RIP machinery. Dp(B362i), which is only approximately 117 kb long, failed to suppress RIP. RIP suppression in gene-sized duplications by large duplications was demonstrated using another test gene, dow, and supposedly applies generally. Crosses homozygous for Dp(AR17) or Dp(IBj5) were as barren as heterozygous crosses. Barrenness of the heterozygous but not the homozygous crosses was suppressible by Sad-1, a semidominant suppressor of RNAi-dependent meiotic silencing by unpaired DNA. A model is proposed in which large duplications recessively suppress semidominant Sad-1 mutations. The wild-isolated Sugartown strain is hypothesized to contain a duplication that confers not only dominant suppression of RIP but also a barren phenotype, which is linked (9%) to supercontig 7.118 in LG VII. PMID- 16361236 TI - The evolution of cytoplasmic incompatibility types: integrating segregation, inbreeding and outbreeding. AB - Cytoplasmic incompatibility (CI) is a reproductive incompatibility induced by maternally transmitted bacteria of the genera Wolbachia and Cardinium. In the simplest form of CI, offspring from infected males and uninfected females suffer from increased mortality. However, it has been noted that crosses between males and females carrying different strains of infection are often also incompatible. The evolutionary processes leading to the emergence of new CI-compatibility types are still not resolved. Here, we develop a model that extends previous theoretical approaches by including segregation of bacterial strains during transmission as well as a continuum of breeding systems ranging from inbreeding (complete sib mating) to outbreeding (complete sib-mating avoidance). Our results demonstrate that (1) with segregation of strains, evolution is unlikely to lead to new CI types that co-occur as a double infection with the preexisting one, (2) inbreeding substantially hampers the evolution of new CI types, and (3) outbreeding facilitates the evolution of new CI types. Our model also provides a hypothesis on the evolutionary origin of CI. PMID- 16361237 TI - Variable pleiotropic effects from mutations at the same locus hamper prediction of fitness from a fitness component. AB - The relationship of genotype, fitness components, and fitness can be complicated by genetic effects such as pleiotropy and epistasis and by heterogeneous environments. However, because it is often difficult to measure genotype and fitness directly, fitness components are commonly used to estimate fitness without regard to genetic architecture. The small bacteriophage X174 enables direct evaluation of genetic and environmental effects on fitness components and fitness. We used 15 mutants to study mutation effects on attachment rate and fitness in six hosts. The mutants differed from our lab strain of X174 by only one or two amino acids in the major capsid protein (gpF, sites 101 and 102). The sites are variable in natural and experimentally evolved X174 populations and affect phage attachment rate. Within the limits of detection of our assays, all mutations were neutral or deleterious relative to the wild type; 11 mutants had decreased host range. While fitness was predictable from attachment rate in most cases, 3 mutants had rapid attachment but low fitness on most hosts. Thus, some mutations had a pleiotropic effect on a fitness component other than attachment rate. In addition, on one host most mutants had high attachment rate but decreased fitness, suggesting that pleiotropic effects also depended on host. The data highlight that even in this simple, well-characterized system, prediction of fitness from a fitness component depends on genetic architecture and environment. PMID- 16361238 TI - The concordance of gene trees and species trees at two linked loci. AB - The gene genealogies of two linked loci in three species are analyzed using a series of Markov chain models. We calculate the probability that the gene tree of one locus is concordant with the species tree, given that the gene tree of the other locus is concordant. We define a threshold value of the recombination rate, r*, to be the rate for which the difference between the conditional probability of concordance and its asymptotic value is reduced to 5% of the initial difference. We find that, although r* depends in a complicated way on the times of speciation and effective population sizes, it is always relatively small, <10/N4, where N4 is the effective size of the species represented by the internal branch of the species tree. Consequently, the concordance of gene trees of neutral loci with the species tree is expected to be on roughly the same length scale on the chromosome as the extent of significant linkage disequilibrium within species unless the effective size of contemporary populations is very different from the effective sizes of their ancestral populations. Both balancing selection and selective sweeps can result in much longer genomic regions having concordant gene trees. PMID- 16361239 TI - Allele frequency distribution under recurrent selective sweeps. AB - The allele frequency of a neutral variant in a population is pushed either upward or downward by directional selection on a linked beneficial mutation ("selective sweeps"). DNA sequences sampled after the fixation of the beneficial allele thus contain an excess of rare neutral alleles. This study investigates the allele frequency distribution under selective sweep models using analytic approximation and simulation. First, given a single selective sweep at a fixed time, I derive an expression for the sampling probabilities of neutral mutants. This solution can be used to estimate the time of the fixation of a beneficial allele from sequence data. Next, I obtain an approximation to mean allele frequencies under recurrent selective sweeps. Under recurrent sweeps, the frequency spectrum is skewed toward rare alleles. However, the excess of high-frequency derived alleles, previously shown to be a signature of single selective sweeps, disappears with recurrent sweeps. It is shown that, using this approximation and multilocus polymorphism data, genomewide parameters of directional selection can be estimated. PMID- 16361240 TI - Clinal variation in phyB2, a candidate gene for day-length-induced growth cessation and bud set, across a latitudinal gradient in European aspen (Populus tremula). AB - The initiation of growth cessation and dormancy represents a critical ecological and evolutionary trade-off between survival and growth in most forest trees. The most important environmental cue regulating the initiation of dormancy is a shortening of the photoperiod and phytochrome genes have been implicated in short day-induced bud set and growth cessation in Populus. We characterized patterns of DNA sequence variation at the putative candidate gene phyB2 in 4 populations of European aspen (Populus tremula) and scored single-nucleotide polymorphisms in an additional 12 populations collected along a latitudinal gradient in Sweden. We also measured bud set from a subset of these trees in a growth chamber experiment. Buds set showed significant clinal variation with latitude, explaining approximately 90% of the population variation in bud set. A sliding window scan of phyB2 identified six putative regions with enhanced population differentiation and four SNPs showed significant clinal variation. The clinal variation at individual SNPs is suggestive of an adaptive response in phyB2 to local photoperiodic conditions. Three of four SNPs showing clinal variation were located in regions with excessive genetic differentiation, demonstrating that searching for regions of high genetic differentiation can be useful for identifying sites putatively involved in local adaptation. PMID- 16361241 TI - Evolutionary rates and expression level in Chlamydomonas. AB - In many biological systems, especially bacteria and unicellular eukaryotes, rates of synonymous and nonsynonymous nucleotide divergence are negatively correlated with the level of gene expression, a phenomenon that has been attributed to natural selection. Surprisingly, this relationship has not been examined in many important groups, including the unicellular model organism Chlamydomonas reinhardtii. Prior to this study, comparative data on protein-coding sequences from C. reinhardtii and its close noninterfertile relative C. incerta were very limited. We compiled and analyzed protein-coding sequences for 67 nuclear genes from these taxa; the sequences were mostly obtained from the C. reinhardtii EST database and our C. incerta EST data. Compositional and synonymous codon usage biases varied among genes within each species but were highly correlated between the orthologous genes of the two species. Relative rates of synonymous and nonsynonymous substitution across genes varied widely and showed a strong negative correlation with the level of gene expression estimated by the codon adaptation index. Our comparative analysis of substitution rates in introns of lowly and highly expressed genes suggests that natural selection has a larger contribution than mutation to the observed correlation between evolutionary rates and gene expression level in Chlamydomonas. PMID- 16361242 TI - Efficient inference of haplotypes from genotypes on a large animal pedigree. AB - We present a simple algorithm for reconstruction of haplotypes from a sample of multilocus genotypes. The algorithm is aimed specifically for analysis of very large pedigrees for small chromosomal segments, where recombination frequency within the chromosomal segment can be assumed to be zero. The algorithm was tested both on simulated pedigrees of 155 individuals in a family structure of three generations and on real data of 1149 animals from the Israeli Holstein dairy cattle population, including 406 bulls with genotypes, but no females with genotypes. The rate of haplotype resolution for the simulated data was >91% with a standard deviation of 2%. With 20% missing data, the rate of haplotype resolution was 67.5% with a standard deviation of 1.3%. In both cases all recovered haplotypes were correct. In the real data, allele origin was resolved for 22% of the heterozygous genotypes, even though 70% of the genotypes were missing. Haplotypes were resolved for 36% of the males. Computing time was insignificant for both data sets. Despite the intricacy of large-scale real pedigree genotypes, the proposed algorithm provides a practical rule-based solution for resolving haplotypes for small chromosomal segments in commercial animal populations. PMID- 16361243 TI - Optimal design and analysis of genetic studies on gene expression. AB - Whole-genome profiling of gene expression in a segregating population has the potential to identify the regulatory consequences of natural allelic variation. Costs of such studies are high and require that resources--microarrays and population--are used as efficiently as possible. We show that current studies can be improved significantly by a new design for two-color microarrays. Our "distant pair design" profiles twice as many individuals as there are arrays, cohybridizes individuals with dissimilar genomes, gives more weight to known regulatory loci if wished, and therewith maximizes the power for decomposing expression variation into regulatory factors. It can also exploit a large population (larger than twice the number of available microarrays) as a useful resource to select the most dissimilar pairs of individuals from. Our approach identifies more regulatory factors than alternative strategies do in computer simulations for realistic genome sizes, and similar promising results are obtained in an application on Arabidopsis thaliana. Our results will aid the design and analysis of future studies on gene expression and will help to shed more light on gene regulatory networks. PMID- 16361244 TI - Modeling haplotype block variation using Markov chains. AB - Models of background variation in genomic regions form the basis of linkage disequilibrium mapping methods. In this work we analyze a background model that groups SNPs into haplotype blocks and represents the dependencies between blocks by a Markov chain. We develop an error measure to compare the performance of this model against the common model that assumes that blocks are independent. By examining data from the International Haplotype Mapping project, we show how the Markov model over haplotype blocks is most accurate when representing blocks in strong linkage disequilibrium. This contrasts with the independent model, which is rendered less accurate by linkage disequilibrium. We provide a theoretical explanation for this surprising property of the Markov model and relate its behavior to allele diversity. PMID- 16361245 TI - Simulating the collaborative cross: power of quantitative trait loci detection and mapping resolution in large sets of recombinant inbred strains of mice. AB - It has been suggested that the collaborative cross, a large set of recombinant inbred strains derived from eight inbred mouse strains, would be a powerful resource for the dissection of complex phenotypes. Here we use simulation to investigate the power of the collaborative cross to detect and map small genetic effects. We show that for a fixed population of 1000 individuals, 500 RI lines bred using a modified version of the collaborative cross design are adequate to map a single additive locus that accounts for 5% of the phenotypic variation to within 0.96 cM. In the presence of strong epistasis more strains can improve detection, but 500 lines still provide sufficient resolution to meet most goals of the collaborative cross. However, even with a very large panel of RILs, mapping resolution may not be sufficient to identify single genes unambiguously. Our results are generally applicable to the design of RILs in other species. PMID- 16361246 TI - Recently evolved genes identified from Drosophila yakuba and D. erecta accessory gland expressed sequence tags. AB - The fraction of the genome associated with male reproduction in Drosophila may be unusually dynamic. For example, male reproduction-related genes show higher-than average rates of protein divergence and gene expression evolution compared to most Drosophila genes. Drosophila male reproduction may also be enriched for novel genetic functions. Our earlier work, based on accessory gland protein genes (Acp's) in D. simulans and D. melanogaster, suggested that the melanogaster subgroup Acp's may be lost and/or gained on a relatively rapid timescale. Here we investigate this possibility more thoroughly through description of the accessory gland transcriptome in two melanogaster subgroup species, D. yakuba and D. erecta. A genomic analysis of previously unknown genes isolated from cDNA libraries of these species revealed several cases of genes present in one or both species, yet absent from ingroup and outgroup species. We found no evidence that these novel genes are attributable primarily to duplication and divergence, which suggests the possibility that Acp's or other genes coding for small proteins may originate from ancestrally noncoding DNA. PMID- 16361247 TI - Elimination of abnormal sialylglycoproteins in fibroblasts with sialidosis and galactosialidosis by normal gene transfer and enzyme replacement. AB - Sialidosis and galactosialidosis are lysosomal storage diseases caused by the genetic defects of lysosomal sialidase (neuraminidase-1; NEU1) and lysosomal protective protein/cathepsin A (PPCA), respectively, associated with a NEU1 deficiency, excessive accumulation of sialylglycoconjugates, and development of progressive neurosomatic manifestations; in addition, the latter disorder is accompanied by simultaneous deficiencies of beta-galactosidase and cathepsin A. We demonstrated that a few soluble N-glycosylated proteins carrying sialyloligosaccharides sensitive to glycopeptidase F (GPF) can be specifically detected in cultured fibroblasts from sialidosis and galactosialidosis cases by blotting with a Maackia amurensis (MAM) lectin. We also examined the therapeutic effects of normal gene transfer and enzyme replacement by evaluating the decreases in sialylglycoconjugates accumulated in fibroblasts with these NEU1 deficiencies. The specific N-glycosylated proteins detected on MAM lectin blotting as well as the granular lysosomal fluorescence due to an avidin FITC/biotinylated MAM lectin conjugate in sialidosis and galactosialidosis fibroblasts disappeared in parallel with the restoration of the intracellular NEU1 activity after transfection of the recombinant NEU1 fused to HA tag sequence and the wild-type PPCA cDNA as well as administration of the recombinant PPCA precursor protein. The detection method for the abnormal sialylglycoproteins in cultured cells involving MAM lectin was demonstrated to be useful not only for biochemical and diagnostic analyses of NEU1 deficiencies but also for therapeutic evaluation of these conditions. PMID- 16361248 TI - Mechanisms of pharmacological rescue of trafficking-defective hERG mutant channels in human long QT syndrome. AB - Long QT syndrome type 2 is caused by mutations in the human ether-a-go-go-related gene (hERG). We previously reported that the N470D mutation is retained in the endoplasmic reticulum (ER) but can be rescued to the plasma membrane by hERG channel blocker E-4031. The mechanisms of ER retention and how E-4031 rescues the N470D mutant are poorly understood. In this study, we investigated the interaction of hERG channels with the ER chaperone protein calnexin. Using coimmunoprecipitation, we showed that the immature forms of both wild type hERG and N470D associated with calnexin. The association required N-linked glycosylation of hERG channels. Pulse-chase analysis revealed that N470D had a prolonged association with calnexin compared with wild type hERG and E-4031 shortened the time course of calnexin association with N470D. To test whether the prolonged association of N470D with calnexin is due to defective folding of mutant channels, we studied hERG channel folding using the trypsin digestion method. We found that N470D and the immature form of wild type hERG were more sensitive to trypsin digestion than the mature form of wild type hERG. In the presence of E-4031, N470D became more resistant to trypsin even when its ER-to Golgi transport was blocked by brefeldin A. These results suggest that defective folding of N470D contributes to its prolonged association with calnexin and ER retention and that E-4031 may restore proper folding of the N470D channel leading to its cell surface expression. PMID- 16361249 TI - Biochemical characterization of the diaphanous autoregulatory interaction in the formin homology protein FHOD1. AB - Diaphanous related formins (DRFs) are cytoskeleton remodeling proteins that mediate specific upstream GTPase signals to regulate cellular processes such as cytokinesis, cell polarity, and organelle motility. Previous work on the Rho interacting DRF mDia has established that the biological activity of DRFs is regulated by an autoinhibitory interaction of a C-terminal diaphanous autoregulatory domain (DAD) with the DRF N terminus. This autoinhibition is released upon competitive binding of an activated GTPase to the N terminus of the DRF. Analyzing autoregulation of the Rac1-interacting DRF FHOD1, we utilized in vitro binding studies to identify a 60-amino acid DAD at the protein C terminus that recognizes an N-terminal formin homology (FH) 3 domain. Importantly, the FH3 domain of FHOD1 does not overlap with the proposed Rac1-binding domain. The FHOD1 DAD was found to contain one functional hydrophobic autoregulatory motif, while a previously uncharacterized basic cluster that is conserved in all DRF family DADs also contributed to the FH3-DAD interaction. Simultaneous mutation of both motifs efficiently released autoinhibition of FHOD1 in NIH3T3 cells resulting in the formation of actin stress fibers and increased serum response element transcription. A second putative hydrophobic autoregulatory motif N-terminal of the DAD belongs to a unique FHOD subdomain of yet undefined function. NMR structural analysis and size exclusion chromatography experiments revealed that the FHOD1 DAD is intrinsically unstructured with a tendency for a helical conformation in the hydrophobic autoregulation motif. Together, these data suggest that in FHOD1, DAD acts as signal sequence for binding to the well folded and monomeric FH3 domain and imply an activation mechanism that differs from competitive binding of Rac1 and DAD to one interaction site. PMID- 16361250 TI - The Saccharomyces cerevisiae EHT1 and EEB1 genes encode novel enzymes with medium chain fatty acid ethyl ester synthesis and hydrolysis capacity. AB - Fatty acid ethyl esters are secondary metabolites produced by Saccharomyces cerevisiae and many other fungi. Their natural physiological role is not known but in fermentations of alcoholic beverages and other food products they play a key role as flavor compounds. Information about the metabolic pathways and enzymology of fatty acid ethyl ester biosynthesis, however, is very limited. In this work, we have investigated the role of a three-member S. cerevisiae gene family with moderately divergent sequences (YBR177c/EHT1, YPL095c/EEB1, and YMR210w). We demonstrate that two family members encode an acyl-coenzymeA:ethanol O-acyltransferase, an enzyme required for the synthesis of medium-chain fatty acid ethyl esters. Deletion of either one or both of these genes resulted in severely reduced medium-chain fatty acid ethyl ester production. Purified glutathione S-transferase-tagged Eht1 and Eeb1 proteins both exhibited acyl coenzymeA:ethanol O-acyltransferase activity in vitro, as well as esterase activity. Overexpression of Eht1 and Eeb1 did not enhance medium-chain fatty acid ethyl ester content, which is probably due to the bifunctional synthesis and hydrolysis activity. Molecular modeling of Eht1 and Eeb1 revealed the presence of a alpha/beta-hydrolase fold, which is generally present in the substrate-binding site of esterase enzymes. Hence, our results identify Eht1 and Eeb1 as novel acyl coenzymeA:ethanol O-acyltransferases/esterases, whereas the third family member, Ymr210w, does not seem to play an important role in medium-chain fatty acid ethyl ester formation. PMID- 16361251 TI - SUMO-3 enhances androgen receptor transcriptional activity through a sumoylation independent mechanism in prostate cancer cells. AB - Androgens are important for male sexual development, which depend on the cognate receptor, the androgen receptor. The transcriptional activity of the androgen receptor, like other nuclear receptors, is regulated by accessory proteins that can have either positive or negative effects. Through a yeast functional screen, we have identified SUMO-3 as a regulator of androgen receptor activity in prostate cancer cells. SUMO-3 is one of three eukaryotic proteins that become post-translationally conjugated to their target proteins in a manner analogous to the attachment of ubiquitin. In primary prostate epithelial cells, PrEC, and the prostate cancer cells, PC-3, SUMO-3 has a weak negative effect on androgen receptor transcriptional activity. In contrast, SUMO-3 and it close relative SUMO 2 strongly enhance transactivation by endogenous androgen receptor in LNCaP cells. This positive effect is observed in both androgen-dependent and androgen independent LNCaP cells. Interestingly, SUMO-1, unlike SUMO-3 and SUMO-2, can inhibit, but not stimulate, androgen receptor activity. Mutational analysis of the androgen receptor and SUMO-3 demonstrates that the SUMO-3-positive activity does not depend on either the sumoylation sites of the androgen receptor or the sumoylation function of SUMO-3. Stable overexpression of SUMO-3 in LNCaP cells significantly enhances the androgen-dependent proliferation of these cells. Additionally, siRNA-mediated repression of SUMO-2 significantly inhibits the growth of both androgen-dependent and -independent LNCaP cells. Collectively, these results suggest (i) a novel mechanism for elevating AR activity through the switch of SUMO-3 from a weak negative regulator in normal prostate cells to a strong positive regulator in prostate cancer cells and (ii) a proliferative role for SUMO-3 and SUMO-2 in the growth of prostate cancer cells that is independent of sumoylation of the androgen receptor. PMID- 16361252 TI - Glycosylphosphatidylinositol-anchored proteins are required for the transport of detergent-resistant microdomain-associated membrane proteins Tat2p and Fur4p. AB - In eukaryotic cells many cell surface proteins are attached to the membrane via the glycosylphosphatidylinositol (GPI) moiety. In yeast, GPI also plays important roles in the production of mannoprotein in the cell wall. We previously isolated gwt1 mutants and found that GWT1 is required for inositol acylation in the GPI biosynthetic pathway. In this study we isolated a new gwt1 mutant allele, gwt1 10, that shows not only high temperature sensitivity but also low temperature sensitivity. The gwt1-10 cells show impaired acyltransferase activity and attachment of GPI to proteins even at the permissive temperature. We identified TAT2, which encodes a high affinity tryptophan permease, as a multicopy suppressor of cold sensitivity in gwt1-10 cells. The gwt1-10 cells were also defective in the import of tryptophan, and a lack of tryptophan caused low temperature sensitivity. Microscopic observation revealed that Tat2p is not transported to the plasma membrane but is retained in the endoplasmic reticulum in gwt1-10 cells grown under tryptophan-poor conditions. We found that Tat2p was not associated with detergent-resistant membranes (DRMs), which are required for the recruitment of Tat2p to the plasma membrane. A similar result was obtained for Fur4p, a uracil permease localized in the DRMs of the plasma membrane. These results indicate that GPI-anchored proteins are required for the recruitment of membrane proteins Tat2p and Fur4p to the plasma membrane via DRMs, suggesting that some membrane proteins are redistributed in the cell in response to environmental and nutritional conditions due to an association with DRMs that is dependent on GPI-anchored proteins. PMID- 16361253 TI - Critical elements of oligosaccharide acceptor substrates for the Pasteurella multocida hyaluronan synthase. AB - Three-dimensional structures are not available for polysaccharide synthases and only minimal information on the molecular basis for catalysis is known. The Pasteurella multocida hyaluronan synthase (PmHAS) catalyzes the polymerization of the alternating beta1,3-N-acetylglucosamine-beta1,4-glucuronic acid sugar chain by the sequential addition of single monosaccharides to the non-reducing terminus. Therefore, PmHAS possesses both GlcNAc-transferase and glucuronic acid (GlcUA)-transferase activities. The recombinant Escherichia coli-derived PmHAS enzyme will elongate exogenously supplied hyaluronan chains in vitro with either a single monosaccharide or a long chain depending on the UDP-sugar availability. Competition studies using pairs of acceptors with distinct termini (where one oligosaccharide is a substrate that may be elongated, whereas the other cannot) were performed here; the lack of competition suggests that PmHAS contains at least two distinct acceptor sites. We hypothesize that the size of the acceptor binding pockets of the enzyme corresponds to the size of the smallest high efficiency substrates; thus we tested the relative activity of a series of authentic hyaluronan oligosaccharides and related structural analogs. The GlcUA transferase site readily elongates (GlcNAc-GlcUA)(2), whereas the GlcNAc transferase elongates GlcUA-Glc-NAc-GlcUA. The minimally sized oligosaccharides, elongated with high efficiency, both contain a trisaccharide with two glucuronic acid residues that enabled the identification of a synthetic, artificial acceptor for the synthase. PmHAS behaves as a fusion of two complete glycosyltransferases, each containing a donor site and an acceptor site, in one polypeptide. Overall, this information advances the knowledge of glycosaminoglycan biosynthesis as well as assists the creation of various therapeutic sugars for medical applications in the future. PMID- 16361254 TI - Naturally occurring mutations in human mitochondrial pre-tRNASer(UCN) can affect the transfer ribonuclease Z cleavage site, processing kinetics, and substrate secondary structure. AB - tRNAs are transcribed as precursors with a 5' end leader and a 3' end trailer. The 5' end leader is processed by RNase P, and in most organisms in all three kingdoms, transfer ribonuclease (tRNase) Z can endonucleolytically remove the 3' end trailer. Long ((L)) and short ((S)) forms of the tRNase Z gene are present in the human genome. tRNase Z(L) processes a nuclear-encoded pre-tRNA approximately 1600-fold more efficiently than tRNase Z(S) and is predicted to have a strong mitochondrial transport signal. tRNase Z(L) could, thus, process both nuclear- and mitochondrially encoded pre-tRNAs. More than 150 pathogenesis-associated mutations have been found in the mitochondrial genome, most of them in the 22 mitochondrially encoded tRNAs. All the mutations investigated in human mitochondrial tRNA(Ser(UCN)) affect processing efficiency, and some affect the cleavage site and secondary structure. These changes could affect tRNase Z processing of mutant pre-tRNAs, perhaps contributing to mitochondrial disease. PMID- 16361255 TI - Combined signaling through ERK, PI3K/AKT, and RAC1/p38 is required for met triggered cortical neuron migration. AB - Cell migration is a complex biological process playing a key role in physiological and pathological conditions. During central nervous system development, positioning and function of cortical neurons is tightly regulated by cell migration. Recently, signaling events involving the urokinase-type plasminogen activator receptor, which is a key regulator for the activation of hepatocyte growth factor (HGF), have been implicated in modulating cortical neuron migration. However, the intracellular pathways controlling neuronal migration triggered by the HGF receptor Met have not been elucidated. By combining pharmacological and genetic approaches, we show here that the Ras/ERK pathway and phosphatidylinositol 3-kinase (PI3K) are both required for cortical neuron migration. By dissecting the downstream signals necessary for this event, we found that Rac1/p38 and Akt are required, whereas the c-Jun N-terminal kinase (JNK) and mTOR/p70(s6k) pathways are dispensable. This study demonstrates that concomitant activation of the Ras/ERK, PI3K/Akt, and Rac1/p38 pathways is required to achieve full capacity of cortical neurons to migrate upon HGF stimulation. PMID- 16361256 TI - TRP-ML1 regulates lysosomal pH and acidic lysosomal lipid hydrolytic activity. AB - Mucolipidosis type IV (MLIV) is caused by mutations in the ion channel mucolipin 1 (TRP-ML1). MLIV is typified by accumulation of lipids and membranous materials in intracellular organelles, which was hypothesized to be caused by the altered membrane fusion and fission events. How mutations in TRP-ML1 lead to aberrant lipolysis is not known. Here we present evidence that MLIV is a metabolic disorder that is not associated with aberrant membrane fusion/fission events. Thus, measurement of lysosomal pH revealed that the lysosomes in TRP-ML1(-/-) cells obtained from the patients with MLIV are over-acidified. TRP-ML1 can function as a H(+) channel, and the increased lysosomal acidification in TRP-ML1( /-) cells is likely caused by the loss of TRP-ML1-mediated H(+) leak. Measurement of lipase activity using several substrates revealed a marked reduction in lipid hydrolysis in TRP-ML1(-/-) cells, which was rescued by the expression of TRP-ML1. Cell fractionation indicated specific loss of acidic lipase activity in TRP-ML1( /-) cells. Furthermore, dissipation of the acidic lysosomal pH of TRP-ML1(-/-) cells by nigericin or chloroquine reversed the lysosomal storage disease phenotype. These findings provide a new mechanism to account for the pathogenesis of MLIV. PMID- 16361257 TI - Occupancy of a single anesthetic binding pocket is sufficient to enhance glycine receptor function. AB - Alcohols and volatile anesthetics enhance the function of inhibitory glycine receptors (GlyRs). This is hypothesized to occur by their binding to a pocket formed between the transmembrane domains of individual alpha1 GlyR subunits. Because GlyRs are pentameric, it follows that each GlyR contains up to five alcohol/anesthetic binding sites, with one in each subunit. We asked how many subunits per pentamer need be bound by drug in order to enhance receptor-mediated currents. A cysteine mutation was introduced at amino acid serine 267 (S267C) in the transmembrane 2 domain as a tool to block GlyR potentiation by some anesthetic drugs and to provide a means for covalent binding by the small, anesthetic-like thiol reagent propyl methanethiosulfonate. Xenopus laevis oocytes were co-injected with various ratios of wild-type (wt) to S267C alpha1 GlyR cDNAs in order to express heteromeric receptors with a range of wt:mutant subunit stoichiometries. The enhancement of GlyR currents by 200 mm ethanol and 1.5 mm chloroform was positively correlated with the number of wt subunits found in heteromeric receptors. Furthermore, currents from oocytes injected with high ratios of wt to S267C cDNAs (up to 200:1) were significantly and irreversibly enhanced following propyl methanethiosulfonate labeling and washout, demonstrating that drug binding to a single subunit in the receptor pentamer is sufficient to induce enhancement of GlyR currents. PMID- 16361258 TI - 6-hydroxydopamine-induced apoptosis is mediated via extracellular auto-oxidation and caspase 3-dependent activation of protein kinase Cdelta. AB - 6-Hydroxydopamine is a neurotoxin commonly used to lesion dopaminergic pathways and generate experimental models for Parkinson disease, however, the cellular mechanism of 6-hydroxydopamine-induced neurodegeneration is not well defined. In this study we have explored how 6-hydroxydopamine neurotoxicity is initiated. We have also investigated downstream signaling pathways activated in response to 6 hydroxydopamine, using a neuronal-like, catecholaminergic cell line (PC12 cells) as an in vitro model system. We have shown that 6-hydroxydopamine neurotoxicity is initiated via extracellular auto-oxidation and the induction of oxidative stress from the oxidative products generated. Neurotoxicity is completely attenuated by preincubation with catalase, suggesting that hydrogen peroxide, at least in part, evokes neuronal cell death in this model. 6-Hydroxydopamine does not initiate toxicity by dopamine transporter-mediated uptake into PC12 cells, because both GBR-12909 and nisoxetine (inhibitors of dopamine and noradrenaline transporters, respectively) failed to reduce toxicity. 6-Hydroxydopamine has previously been shown to induce both apoptotic and necrotic cell-death mechanisms. In this study oxidative stress initiated by 6-hydroxydopamine caused mitochondrial dysfunction, activation of caspases 3/7, nuclear fragmentation, and apoptosis. We have shown that, in this model, proteolytic activation of the proapoptotic protein kinase Cdelta (PKCdelta) is a key mediator of 6 hydroxydopamine-induced cell death. 6-Hydroxydopamine induces caspase 3-dependent cleavage of full-length PKCdelta (79 kDa) to yield a catalytic fragment (41 kDa). Inhibition of PKCdelta (with rottlerin or via RNA interference-mediated gene suppression) ameliorates the neurotoxicity evoked by 6-hydroxydopamine, implicating this kinase in 6-hydroxydopamine-induced neurotoxicity and Parkinsonian neurodegeneration. PMID- 16361259 TI - Nucleotide-binding domains of cystic fibrosis transmembrane conductance regulator, an ABC transporter, catalyze adenylate kinase activity but not ATP hydrolysis. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel in the ATP-binding cassette (ABC) transporter family. CFTR consists of two transmembrane domains, two nucleotide-binding domains (NBD1 and NBD2), and a regulatory domain. Previous biochemical reports suggest NBD1 is a site of stable nucleotide interaction with low ATPase activity, whereas NBD2 is the site of active ATP hydrolysis. It has also been reported that NBD2 additionally possessed adenylate kinase (AK) activity. Knowledge about the intrinsic biochemical activities of the NBDs is essential to understanding the Cl(-) ion gating mechanism. We find that purified mouse NBD1, human NBD1, and human NBD2 function as adenylate kinases but not as ATPases. AK activity is strictly dependent on the addition of the adenosine monophosphate (AMP) substrate. No liberation of [(33)P]phosphate is observed from the gamma-(33)P-labeled ATP substrate in the presence or absence of AMP. AK activity is intrinsic to both human NBDs, as the Walker A box lysine mutations abolish this activity. At low protein concentration, the NBDs display an initial slower nonlinear phase in AK activity, suggesting that the activity results from homodimerization. Interestingly, the G551D gating mutation has an exaggerated nonlinear phase compared with the wild type and may indicate this mutation affects the ability of NBD1 to dimerize. hNBD1 and hNBD2 mixing experiments resulted in an 8-57-fold synergistic enhancement in AK activity suggesting heterodimer formation, which supports a common theme in ABC transporter models. A CFTR gating mechanism model based on adenylate kinase activity is proposed. PMID- 16361260 TI - Targeting amyloid-beta peptide (Abeta) oligomers by passive immunization with a conformation-selective monoclonal antibody improves learning and memory in Abeta precursor protein (APP) transgenic mice. AB - Passive immunization of murine models of Alzheimer disease amyloidosis reduces amyloid-beta peptide (Abeta) levels and improves cognitive function. To specifically address the role of Abeta oligomers in learning and memory, we generated a novel monoclonal antibody, NAB61, that preferentially recognizes a conformational epitope present in dimeric, small oligomeric, and higher order Abeta structures but not full-length amyloid-beta precursor protein or C-terminal amyloid-beta precursor protein fragments. NAB61 also recognized a subset of brain Abeta deposits, preferentially mature senile plaques, and amyloid angiopathy. Using NAB61 as immunotherapy, we showed that aged Tg2576 transgenic mice treated with NAB61 displayed significant improvements in spatial learning and memory relative to control mice. These data implicated Abeta oligomers as a pathologic substrate for cognitive decline in Alzheimer disease. PMID- 16361261 TI - Hypothyroid phenotype is contributed by mitochondrial complex I inactivation due to translocated neuronal nitric-oxide synthase. AB - Although transcriptional effects of thyroid hormones have substantial influence on oxidative metabolism, how thyroid sets basal metabolic rate remains obscure. Compartmental localization of nitric-oxide synthases is important for nitric oxide signaling. We therefore examined liver neuronal nitric-oxide synthase-alpha (nNOS) subcellular distribution as a putative mechanism for thyroid effects on rat metabolic rate. At low 3,3',5-triiodo-L-thyronine levels, nNOS mRNA increased by 3-fold, protein expression by one-fold, and nNOS was selectively translocated to mitochondria without changes in other isoforms. In contrast, under thyroid hormone administration, mRNA level did not change and nNOS remained predominantly localized in cytosol. In hypothyroidism, nNOS translocation resulted in enhanced mitochondrial nitric-oxide synthase activity with low O2 uptake. In this context, NO utilization increased active O2 species and peroxynitrite yields and tyrosine nitration of complex I proteins that reduced complex activity. Hypothyroidism was also associated to high phospho-p38 mitogen-activated protein kinase and decreased phospho-extracellular signal-regulated kinase 1/2 and cyclin D1 levels. Similarly to thyroid hormones, but without changing thyroid status, nitric-oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester increased basal metabolic rate, prevented mitochondrial nitration and complex I derangement, and turned mitogen-activated protein kinase signaling and cyclin D1 expression back to control pattern. We surmise that nNOS spatial confinement in mitochondria is a significant downstream effector of thyroid hormone and hypothyroid phenotype. PMID- 16361262 TI - Protein kinase Ciota promotes nicotine-induced migration and invasion of cancer cells via phosphorylation of micro- and m-calpains. AB - Nicotine is a major component in cigarette smoke that activates the growth promoting pathways to facilitate the development of lung cancer. However, it is not clear whether nicotine affects cell motility to facilitate tumor metastasis. Here we discovered that nicotine potently induces phosphorylation of both mu- and m-calpains via activation of protein kinase Ciota (PKCiota), which is associated with accelerated migration and invasion of human lung cancer cells. Purified PKCiota directly phosphorylates mu- and m-calpains in vitro. Overexpression of PKCiota results in increased phosphorylation of both mu- and m-calpains in vivo. Nicotine also induces activation of c-Src, which is a known PKCiota upstream kinase. Treatment of cells with the alpha(7) nicotinic acetylcholine receptor inhibitor alpha-bungarotoxin can block nicotine-induced calpain phosphorylation with suppression of calpain activity, wound healing, cell migration, and invasion, indicating that nicotine-induced calpain phosphorylation occurs, at least in part, through a signaling pathway involving the upstream alpha(7) nicotinic acetylcholine receptor. Intriguingly, depletion of PKCiota by RNA interference suppresses nicotine-induced calpain phosphorylation, calpain activity, cell migration, and invasion, indicating that PKCiota is a necessary component in nicotine-mediated cell motility signaling. Importantly, nicotine potently induces secretion of mu- and m-calpains from lung cancer cells into culture medium, which may have potential to cleave substrates in the extracellular matrix. These findings reveal a novel role for PKCiota as a nicotine-activated, physiological calpain kinase that directly phosphorylates and activates calpains, leading to enhanced migration and invasion of human lung cancer cells. PMID- 16361263 TI - Cyanobacterial ClpC/HSP100 protein displays intrinsic chaperone activity. AB - HSP100 proteins are molecular chaperones that belong to the broader family of AAA+ proteins (ATPases associated with a variety of cellular activities) known to promote protein unfolding, disassembly of protein complexes and translocation of proteins across membranes. The ClpC form of HSP100 is an essential, highly conserved, constitutively expressed protein in cyanobacteria and plant chloroplasts, and yet little is known regarding its specific activity as a molecular chaperone. To address this point, ClpC from the cyanobacterium Synechococcus elongatus (SyClpC) was purified using an Escherichia coli-based overexpression system. Recombinant SyClpC showed basal ATPase activity, similar to that of other types of HSP100 protein in non-photosynthetic organisms but different to ClpC in Bacillus subtilis. SyClpC also displayed distinct intrinsic chaperone activity in vitro, first by preventing aggregation of unfolded polypeptides and second by resolubilizing and refolding aggregated proteins into their native structures. The refolding activity of SyClpC was enhanced 3-fold in the presence of the B. subtilis ClpC adaptor protein MecA. Overall, the distinctive ClpC protein in photosynthetic organisms indeed functions as an independent molecular chaperone, and it is so far unique among HSP100 proteins in having both "holding" and disaggregase chaperone activities without the need of other chaperones or adaptor proteins. PMID- 16361264 TI - Iterative in vivo assembly of large and complex transgenes by combining the activities of phiC31 integrase and Cre recombinase. AB - We have used the phiC31 integrase to introduce large DNA sequences into a vertebrate genome and measure the efficiency of integration of intact DNA as a function of insert size. Inserts of 110 kb and 140 kb in length may be integrated with about 25% and 10% efficiency respectively. In order to overcome the problems of constructing transgenes longer than approximately 150 kb we have established a method that we call; 'Iterative Site Specific Integration' (ISSI). ISSI combines the activities of phiC31 integrase and Cre recombinase to enable the iterative and serial integration of transgenic DNA sequences. In principle the procedure may be repeated an arbitrary number of times and thereby allow the integration of tracts of DNA many hundreds of kilobase pairs long. In practice it may be limited by the time needed to check the accuracy of integration at each step of the procedure. We describe two ISSI experiments, in one of which we have constructed a complex array of vertebrate centromeric sequences of 150 kb in size. The principle that underlies ISSI is applicable to transgenesis in all organisms. ISSI may thus facilitate the reconstitution of biosynthetic pathways encoded by many different genes in transgenic plants, the assembly of large vertebrate loci as transgenes and the synthesis of complete genomes in bacteria. PMID- 16361265 TI - Human and mouse oligonucleotide-based array CGH. AB - Array-based comparative genomic hybridization is a high resolution method for measuring chromosomal copy number changes. Here we present a validated protocol using in-house spotted oligonucleotide libraries for array comparative genomic hybridization (CGH). This oligo array CGH platform yields reproducible results and is capable of detecting single copy gains, multi-copy amplifications as well as homozygous and heterozygous deletions as small as 100 kb with high resolution. A human oligonucleotide library was printed on amine binding slides. Arrays were hybridized using a hybstation and analysed using BlueFuse feature extraction software, with >95% of spots passing quality control. The protocol allows as little as 300 ng of input DNA and a 90% reduction of Cot-1 DNA without compromising quality. High quality results have also been obtained with DNA from archival tissue. Finally, in addition to human oligo arrays, we have applied the protocol successfully to mouse oligo arrays. We believe that this oligo-based platform using 'off-the-shelf' oligo libraries provides an easy accessible alternative to BAC arrays for CGH, which is cost-effective, available at high resolution and easily implemented for any sequenced organism without compromising the quality of the results. PMID- 16361266 TI - A novel experimental approach for systematic identification of box H/ACA snoRNAs from eukaryotes. AB - Box H/ACA snoRNAs represent an abundant group of small non-coding RNAs mainly involved in the pseudouridylation of rRNAs and/or snRNAs in eukaryotes and Archaea. In this study, we describe a novel experimental method for systematic identification of box H/ACA snoRNAs from eukaryotes. In the specialized cDNA libraries constructed by this method with total cellular RNAs from human blood cells, the high efficiency of cloning for diverse box H/ACA snoRNAs was achieved and seven novel species of this snoRNA family were identified from human for the first time. Furthermore, the novel method has been successfully applied for the identification of the box H/ACA snoRNAs from Drosophila and the fission yeast, demonstrating a powerful ability for systematic analysis of box H/ACA snoRNAs in a broad spectrum of eukaryotes. PMID- 16361267 TI - Mycobacterium tuberculosis NAD+-dependent DNA ligase is selectively inhibited by glycosylamines compared with human DNA ligase I. AB - DNA ligases are important enzymes which catalyze the joining of nicks between adjacent bases of double-stranded DNA. NAD+-dependent DNA ligases (LigA) are essential in bacteria and are absent in humans. They have therefore been identified as novel, validated and attractive drug targets. Using virtual screening against an in-house database of compounds and our recently determined crystal structure of the NAD+ binding domain of the Mycobacterium tuberculosis LigA, we have identified N1, N(n)-bis-(5-deoxy-alpha-D-xylofuranosylated) diamines as a novel class of inhibitors for this enzyme. Assays involving M.tuberculosis LigA, T4 ligase and human DNA ligase I show that these compounds specifically inhibit LigA from M.tuberculosis. In vitro kinetic and inhibition assays demonstrate that the compounds compete with NAD+ for binding and inhibit enzyme activity with IC50 values in the microM range. Docking studies rationalize the observed specificities and show that among several glycofuranosylated diamines, bis xylofuranosylated diamines with aminoalkyl and 1, 3-phenylene carbamoyl spacers mimic the binding modes of NAD+ with the enzyme. Assays involving LigA-deficient bacterial strains show that in vivo inhibition of ligase by the compounds causes the observed antibacterial activities. They also demonstrate that the compounds exhibit in vivo specificity for LigA over ATP dependent ligase. This class of inhibitors holds out the promise of rational development of new anti-tubercular agents. PMID- 16361268 TI - Dynamic regulation of replication independent deposition of histone H3 in fission yeast. AB - Recently, a histone H3 variant in Drosophila and humans, the H3.3 protein, was shown to replace canonical H3 in active chromatin in a replication-independent (RI) manner. In the fission yeast Schizosaccharomyces pombe, there exists a single form of H3, which is equivalent to H3.3 and is thought to participate in both replication-independent (RI) and replication-coupled (RC) nucleosome assembly. In this study, we show that RI deposition of H3 at heterochromatic regions is consistently lower than that at a gene-free euchromatic region, and deletion of the conserved heterochromatin-specific proteins Swi6 or Clr4 markedly increases RI deposition at heterochromatic regions such as the silent mating-type loci or centromeres. These results clearly show that RI deposition of H3 occurs preferentially in euchromatic regions. We also observed that RI deposition of H3 could be increased at the thi3(+) gene when transcription is induced, indicating transcription further facilitates RI deposition of H3. Taken together, these observations demonstrate that selective deposition of histone H3.3 at transcriptionally active chromatin by the RI assembly pathway is conserved in fission yeast and, thus, our data support an essential role of histone H3 replacement in maintaining active chromatin among diverse eukaryotic organisms ranging from fission yeast to humans. PMID- 16361269 TI - DNA base flipping by a base pair-mimic nucleoside. AB - On the basis of non-covalent bond interactions in nucleic acids, we synthesized the deoxyadenosine derivatives tethering a phenyl group (X) and a naphthyl group (Z) by an amide linker, which mimic a Watson-Crick base pair. Circular dichroism spectra indicated that the duplexes containing X and Z formed a similar conformation regardless of the opposite nucleotide species (A, G, C, T and an abasic site analogue F), which was not observed for the natural duplexes. The values among the natural duplexes containing the A/A, A/G, A/C, A/T and A/F pairs differed by 5.2 kcal mol(-1) while that among the duplexes containing X or Z in place of the adenine differed by only 1.9 or 2.8 kcal mol(-1), respectively. Fluorescence quenching experiments confirmed that 2-amino purine opposite X adopted an unstacked conformation. The structural and thermodynamic analyses suggest that the aromatic hydrocarbon group of X and Z intercalates into a double helix, resulting in the opposite nucleotide base flipping into an unstacked position regardless of the nucleotide species. This observation implies that modifications at the aromatic hydrocarbon group and the amide linker may expand the application of the base pair-mimic nucleosides for molecular biology and biotechnology. PMID- 16361270 TI - Automatic assessment of alignment quality. AB - Multiple sequence alignments play a central role in the annotation of novel genomes. Given the biological and computational complexity of this task, the automatic generation of high-quality alignments remains challenging. Since multiple alignments are usually employed at the very start of data analysis pipelines, it is crucial to ensure high alignment quality. We describe a simple, yet elegant, solution to assess the biological accuracy of alignments automatically. Our approach is based on the comparison of several alignments of the same sequences. We introduce two functions to compare alignments: the average overlap score and the multiple overlap score. The former identifies difficult alignment cases by expressing the similarity among several alignments, while the latter estimates the biological correctness of individual alignments. We implemented both functions in the MUMSA program and demonstrate the overall robustness and accuracy of both functions on three large benchmark sets. PMID- 16361271 TI - Efficient execution of cell death in non-glycolytic cells requires the generation of ROS controlled by the activity of mitochondrial H+-ATP synthase. AB - There is a large body of clinical data documenting that most human carcinomas contain reduced levels of the catalytic subunit of the mitochondrial H+-ATP synthase. In colon and lung cancer this alteration correlates with a poor patient prognosis. Furthermore, recent findings in colon cancer cells indicate that downregulation of the H+-ATP synthase is linked to the resistance of the cells to chemotherapy. However, the mechanism by which the H+-ATP synthase participates in cancer progression is unknown. In this work, we show that inhibitors of the H+ ATP synthase delay staurosporine (STS)-induced cell death in liver cells that are dependent on oxidative phosphorylation for energy provision whereas it has no effect on glycolytic cells. Efficient execution of cell death requires the generation of reactive oxygen species (ROS) controlled by the activity of the H+ ATP synthase in a process that is concurrent with the rapid disorganization of the cellular mitochondrial network. The generation of ROS after STS treatment is highly dependent on the mitochondrial membrane potential and most likely caused by reverse electron flow to Complex I. The generated ROS promote the carbonylation and covalent modification of cellular and mitochondrial proteins. Inhibition of the activity of the H+-ATP synthase blunted ROS production prevented the oxidation of cellular proteins and the modification of mitochondrial proteins delaying the release of cytochrome c and the execution of cell death. The results in this work establish the downregulation of the H+-ATP synthase, and thus of oxidative phosphorylation, as part of the molecular strategy adapted by cancer cells to avoid ROS-mediated cell death. Furthermore, the results provide a mechanistic explanation to understand chemotherapeutic resistance of cancer cells that rely on glycolysis as the main energy provision pathway. PMID- 16361272 TI - Prostaglandin-endoperoxide synthase 2 (PTGS2) gene polymorphisms and risk of biliary tract cancer and gallstones: a population-based study in Shanghai, China. AB - There is evidence that chronic inflammation predisposes to biliary tract cancer and that use of non-steroidal anti-inflammatory drugs (NSAIDs) is protective. Although the mechanisms by which NSAIDs lower cancer risk remain unclear, NSAIDs reduce prostaglandin production by blocking prostaglandin-endoperoxide synthase 2 (PTGS2, commonly known as COX-2), an enzyme induced by proinflammatory stimuli that is often overexpressed in malignant tissue. Since variants in the PTGS2 gene may modify the expression or function of its encoded enzyme to modulate the inflammatory response in the biliary tract, we examined the associations of eight PTGS2 polymorphisms (-645C-->T; Ex3 -8G-->C; IVS5 -275T-->G; IVS7 +111T-->C; Ex10 +127T-->C; Ex10 +686 --->ATTAT-->TTATA; Ex10 +837T-->C; Ex10 -90C-->T) with biliary tract cancer and stones in a population-based case-control study conducted in Shanghai, China. Genotyping was performed for 411 patients with biliary tract cancer (237 gallbladder, 127 extrahepatic bile duct and 47 ampulla of Vater), 895 patients with biliary stones (673 gallbladder, 222 bile duct), and 786 healthy individuals randomly selected from the population. Significant associations were seen only between the Ex10 +837T-->C marker and bile duct cancer risk. Relative to individuals with the TT genotype, those carrying the C allele (TC or CC genotype) had a 1.8-fold (95% confidence interval: 1.2-2.7) risk of bile duct cancer. Inferred haplotypes including this risk-conferring allele were also associated with increased bile duct cancer risk of similar magnitude. Our results suggest that a common PTGS2 variant increases bile duct cancer risk. Further investigation is needed to confirm and extend our findings in studies of biliary tract cancer that more comprehensively examine PTGS2 and other inflammation-related genes. PMID- 16361273 TI - Effects of folate deficiency on gene expression in the apoptosis and cancer pathways in colon cancer cells. AB - Folate is a B vitamin, deficiency of which appears to increase the risk of developing several malignancies including colorectal cancer. In contrast to the cancer-promoting effect of folate deficiency in normal tissues, several lines of evidence indicate that folate depletion suppresses the progression of existing neoplasms and enhance the sensitivity of cancer cells to chemotherapy. Folate mediates the transfer of one-carbon necessary for the de novo biosynthesis of purines and thymidylate, and hence is an essential factor for DNA synthesis and repair, and the maintenance of DNA integrity and stability. Folate deficiency induces DNA strand breaks, increases uracil misincorporation into DNA, impairs DNA repair and appears to induce apoptosis. Although the effects of folate depletion on DNA integrity and apoptosis and on subsequent cancer development, progression and treatment in colonic epithelial cells have been well characterized, it is largely unknown at present how folate depletion modulates specific upstream genes in apoptosis and cancer pathways that regulate these processes. We therefore investigated the effects of folate depletion on expression of genes involved in apoptosis and cancer pathways in four human colon adenocarcinoma cell lines in an in vitro model of folate deficiency. Apoptosis and cancer pathway-specific mini-microarray were used to screen for differentially expressed genes in response to folate deficiency, and the expression of seven most notably and consistently affected genes was confirmed by real time RT-PCR. Our data suggest that folate deficiency affects the expression of key genes that are related to cell cycle control, DNA repair, apoptosis and angiogenesis in a cell-specific manner. Cell-specificity in gene expression changes in response to folate deficiency is likely due to significant differences in molecular and phenotypic characteristics, growth rates and intracellular folate concentrations among the four cell lines. PMID- 16361274 TI - Locally constrained mixture representation of dynamic imaging data from PET and MR studies. AB - Dynamic positron emission tomography (PET) studies provide measurements of the kinetics of radiotracers in living tissue. This is a powerful technology which can play a major role in the study of biological processes, potentially leading to better understanding and treatment of disease. Dynamic PET data relate to complex spatiotemporal processes and its analysis poses significant challenges. In previous work, mixture models that expressed voxel-level PET time course data as a convex linear combination of a finite number of dominant time course characteristics (called sub-TACs) were introduced. This paper extends that mixture model formulation to allow for a weighted combination of scaled sub-TACs and also considers the imposition of local constraints in the number of sub-TACs that can be active at any one voxel. An adaptive 3D scaled segmentation algorithm is developed for model initialization. Increases in the weighted residual sums of squares is used to guide the choice of the number of segments and the number of sub-TACs in the final mixture model. The methodology is applied to five data sets from representative PET imaging studies. The methods are also applicable to other contexts in which dynamic image data are acquired. To illustrate this, data from an echo-planar magnetic resonance (MR) study of cerebral hemodynamics are considered. Our analysis shows little indication of departure from a locally constrained mixture model representation with at most two active components at any voxel. Thus, the primary sources of spatiotemporal variation in representative dynamic PET and MR imaging studies would appear to be accessible to a substantially simplified representation in terms of the generalized locally constrained mixture model introduced. PMID- 16361275 TI - Replication of association of IL1 gene complex members with ankylosing spondylitis in Taiwanese Chinese. AB - OBJECTIVE: To test the association of interleukin 1 (IL1) gene family members with ankylosing spondylitis (AS), previously reported in Europid subjects, in an ethnically remote population. METHODS: 200 Taiwanese Chinese AS patients and 200 ethnically matched healthy controls were genotyped for five single nucleotide polymorphisms (SNPs) and the IL1RN.VNTR, markers previously associated with AS. Allele, genotype, and haplotype frequencies were compared between cases and controls. RESULTS: Association of alleles and genotypes of the markers IL1F10.3, IL1RN.4, and IL1RN.VNTR was observed with AS (p<0.05). Haplotypes of pairs of these markers and of the markers IL1RN.6/1 and IL1RN.6/2 were also significantly associated with AS. The strongest associations observed were with the marker IL1RN.4, and with the two-marker haplotype IL1RN.4-IL1RN.VNTR (both p = 0.004). Strong linkage disequilibrium was observed between all marker pairs except those involving IL1B-511 (D' 0.4 to 0.9, p<0.01). CONCLUSIONS: The IL1 gene cluster is associated with AS in Taiwanese Chinese. This finding provides strong statistical support that the previously observed association of this gene cluster with AS is a true positive finding. These authors contributed equally to the study. PMID- 16361276 TI - Positive anti-cyclic citrullinated proteins and rheumatoid factor during active lung tuberculosis. AB - OBJECTIVES: To determine the prevalence of anti-cyclic citrullinated proteins (anti-CCP) and IgM rheumatoid factor (RF) in sera of patients with TB compared with healthy controls. PATIENTS AND METHODS: 47 consecutive patients with recently diagnosed active pulmonary TB and 39 healthy controls were studied. Data were collected by questionnaire on clinical features of the disease, duration of symptoms, fever, cough, arthralgia, myalgia, sicca symptoms. Serum samples were collected from patients before starting treatment for TB and frozen at -20 degrees C. Anti-CCP and IgM RF were evaluated by ELISA. RESULTS: The mean (SD) duration of TB related symptoms was 4.4 (1.7) months, 73% had fever, 94% a cough. Rheumatic symptoms were relatively rare: arthralgia (4%), myalgias (4%), eye and mouth dryness (2% and 9%, respectively). Mean (SD) levels of anti-CCP were significantly increased in patients with TB compared with controls: 44.9 (51) IU v 20 (7.3) IU (p = 0.002). Serum levels >40 U were found in 15/47 (32%) patients compared with 1/39 (2.6%) controls (p = 0.002). Mean (SD) serum levels of IgM RF were significantly increased in patients with TB: 17.8 (19) v 4.3 (5) (p<0.0001). IgM RF was positive (>6 IU) in 29/47 (62%) patients v 1/39 (2.6%) controls (p<0.0001). CONCLUSIONS: A significant proportion of patients with active TB have an increased titre of anti-CCP and IgM RF. PMID- 16361277 TI - Infliximab leads to a rapid but transient improvement in endothelial function in patients with primary systemic vasculitis. AB - OBJECTIVE: To assess the immediate effects of tumour necrosis factor alpha (TNFalpha) blockade on endothelial function in systemic vasculitis. METHODS: Endothelial function was assessed by laser Doppler flowmetry in patients with active vasculitis after 10 infusions of infliximab. For comparison endothelial responses were assessed after five infusions of cyclophosphamide plus methylprednisolone. RESULTS: Endothelial dependent vasodilatation (EDV) improved significantly within 24 hours of infliximab infusion. The median change in red blood cell flux (interquartile range) was 5.7 (4.3-8.2) before infusion v 8.4 (7.5-10.9) at 24 hours; p=0.027. This was not maintained at day 14. No improvement was seen in EDV after cyclophosphamide plus methylprednisolone infusion. CONCLUSION: The rapid but transient improvement in EDV after TNFalpha inhibition suggests that TNFalpha may have a direct role in the impairment of endothelial function. PMID- 16361278 TI - Conversion from calcineurin inhibitors to sirolimus in chronic allograft nephropathy: benefits and risks. PMID- 16361279 TI - First evidence of fatal hantavirus nephropathy in India, mimicking leptospirosis. PMID- 16361280 TI - Navigating the quagmire: the regulation of human embryonic stem cell research. AB - Embryonic stem (ES) cell research has garnered almost unprecedented attention. Debate over the boundaries of such research is ongoing, and the regulation of the field varies widely between countries. This article identifies and evaluates the four major positions that emanate from current international regulations. ES cell policies may ultimately impact on public health, and hence they must be both rigorous and transparent. We contend that these goals will only be achieved if policy is both ethically consistent and clinically realistic with regard to the ability to achieve therapeutic goals. We conclude that policies allowing the ongoing extraction of stem cells from spare in vitro fertilization embryos and the creation of embryos for research (within set limitations) cope most adequately with the tension between varying views on the moral status of the human embryo and the therapeutic potential inherent within ES cell research. PMID- 16361281 TI - Assisted procreation in cases of hepatitis B, hepatitis C or human immunodeficiency virus infection of the male partner. AB - Guidelines for assisted procreation impose a special responsibility upon physicians for the health of the expected child because of their active role in inducing pregnancy. Therefore, careful clinical evaluation of both partners has to precede every application of these methods. Risks for the mother's health or the development of the child count as a relative contraindication for a treatment. To balance these relative contraindications, the existing risk factors have to be recognized through screening examination. If a chronic infection occurs in the male partner, prevention for the female partner is theoretically possible by using a condom. As this inhibits a pregnancy, at least in cases of human immunodeficiency virus and hepatitis C virus infections, realization of a pregnancy requires assisted procreation. The main question in these cases is whether infectious particles can be eliminated by sperm processing to ensure the safe treatment of the healthy female partner. PMID- 16361282 TI - Long-term conservative therapy for endometrial adenocarcinoma in young women. AB - BACKGROUND: To evaluate the efficacy and safety of long-term conservative therapy with medroxyprogesterone acetate (MPA) for endometrial carcinoma in young patients who had experienced failure after initial therapy or relapse, we reviewed the clinical and pathologic records of eight patients diagnosed with well-differentiated endometrial adenocarcinoma without myometrial invasion who were treated with MPA for over 6 months because of treatment failure or relapse. RESULTS: The average duration of MPA treatment was 22 months. All patients were followed-up for a mean of 76.5 months. Seven patients responded to initial MPA treatment within a period of 14 months (mean, 7.9 months). All these patients experienced relapse and the mean time to relapse was 11.6 months (range, 4-33 months). All six patients with relapse were treated with additional treatments of MPA, and all but one responded to this treatment within a period of 16 months (mean, 8.0 months). Six patients ultimately underwent hysterectomy. All presented well-differentiated endometrioid adenocarcinomas without extrauterine disease. Three became pregnant and two delivered full-term normal infants. No patient died of the disease. CONCLUSION: Although lesions are expected to disappear with prolonged MPA treatment, this form of progestin therapy is hazardous because recurrence occurs frequently. Only strictly selected patients should therefore be indicated for long-term MPA treatment and careful evaluation before and after treatment should be performed. PMID- 16361284 TI - Influence of intermediate and uninterrupted FMR1 CGG expansions in premature ovarian failure manifestation. AB - BACKGROUND: Studies attempting to precisely define the range of fragile mental retardation 1 (FMR1) expansions and its inf luence in premature ovarian failure (POF) manifestation are partially lacking. To this aim, we evaluated a large cohort of POF patients for the size and, in selected cases, for the sequence of the CGG expansion. Furthermore, the correlation between POF and X-inactivation was investigated in FRAXA families. METHODS: By fluorescent PCR, 190 POF and 200 control women were sized for the CGG tract; some subjects were also characterized by sequencing and for the FMR1 activation ratio. RESULTS AND CONCLUSION: We found a significant association (19/190, 10%, P < 1 x 10(-6)) between POF and FMR1 premutation (range 63-163 repeats) and a significant enrichment (9/190, 4.7%, P = 0.021) of POF carriers of intermediate expansions (range 41-58 repeats). Interestingly, intermediate alleles were entirely composed of CGG repeats. Furthermore, the analysis of three pairs of siblings with similar FMR1 expansions and discordant for the POF phenotype showed a direct correlation between the expression of the intermediate/premutated allele and POF manifestation. The results obtained strengthen the correlation between FMR1 expansion and POF and suggest that the manifestation of the ovarian dysfunction could be influenced both by the pattern of interruption of the CGG repeat and by X-inactivation. PMID- 16361283 TI - Cryptorchidism in LhrKO animals and the effect of testosterone-replacement therapy. AB - BACKGROUND: The objective of the current study was to characterize the morphological and genetic basis of cryptorchidism. METHODS AND RESULTS: We investigated cryptorchidism in LH receptor (Lhr) knockout (LhrKO) mice and how testosterone-replacement therapy (TRT) worked to correct the phenotype. The results revealed that while gubernacular development was indistinguishable between Lhr-null and wild-type animals until 7 days of age, it was subsequently severely impaired in null animals. This was due to a reduction in mesenchymal cell division, differentiation into cremaster muscle cells and their delayed maturation. While transcript levels of Hoxa10, Hoxa11, Desrt and Dll1 were indistinguishable, the levels of Notch1, Numb and Lgr8 in the gubernaculum and Insl3 in the testes were lower in Lhr-null than in wild-type siblings. The TRT, which completed testicular descent into the scrotum, corrected the morphological changes and the expression of Lgr8, Numb and Notch, but not Insl3, to wild-type levels. Transection of the genitofemoral nerve did not prevent the TRT effect. CONCLUSION: In summary, cryptorchidism in Lhr-null animals was caused by defects in the gubernacular development due to testosterone deficiency. TRT reversed all the morphological and gene expression changes except Insl3, suggesting that testosterone, not INSL3, secreted by Leydig cells, facilitates the completion of testicular descent. PMID- 16361285 TI - Embryonic stem-cell gametes: the new frontier in human reproduction. AB - As infertility increases and gamete donations decline, an alternate source of sex cells may prove valuable for research and infertility treatment. This article examines the social and scientific value of gametes derived from the differentiation of established human embryonic stem (ES)-cell lines (ES-cell derived gametes) and customized gametes created using nuclear transfer technologies to contain a haploid set of genes creating children genetically related to parent(s). ES-cell-derived gametes may be valuable as a resource for biomedical research, instruction and training in assisted reproductive technologies and perhaps for creating children. The creation of children by ES cell-derived and customized gametes may not result in psychological harm to children but customized gametes may lead to physical harm to children or an accumulation of gene mutations in a population. Although the creation of new types of children using ES-cell gametes provides more reproductive choices to both fertile and infertile individuals, the risk or physical harm to children from customized gametes may be so severe that the scope of reproductive liberty must be limited. Further scientific and ethical analysis of the creation of children by ES-cell gametes is required. PMID- 16361286 TI - Increased sperm DNA damage in patients with varicocele: relationship with seminal oxidative stress. AB - BACKGROUND: The pathophysiology of the testicular damage in varicocele has not been completely understood. Oxidative stress and related sperm DNA damage have been identified as significant causes of male infertility. The current study was designed to determine the extent of sperm nuclear DNA damage in patients with varicocele and to examine its relationship with oxidative stress. METHODS: Semen samples from 55 patients with clinical varicocele and 25 normozoospermic donors were examined. Varicocele sperm samples were classified as normal or abnormal according to World Health Organization guidelines. Sperm DNA damage was evaluated by the sperm chromatin structure assay/flow cytometry and by the terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay. Levels of reactive oxygen species (ROS) and total antioxidant capacity were assessed by a chemiluminescence assay. RESULTS: DNA fragmentation index (DFI) (percentage of sperm with denatured DNA) values and the percentage of TUNEL positive cells were significantly greater in patients with varicocele, either with normal (DFI, 20.7 +/- 4.0; TUNEL positive, 26.1 +/- 3.2) or with abnormal (DFI, 35.5 +/- 9.0; TUNEL positive, 32.2 +/- 4.1) semen profile, compared with controls (DFI, 7.1 +/- 0.9; TUNEL positive, 14.2 +/- 1.2). Similarly, ROS levels were significantly higher (P < 0.01) in both groups of patients with varicocele. CONCLUSIONS: The presence of a varicocele is associated with high levels of DNA damage spermatozoa even in the presence of normal semen profile. The results also indicate that oxidative damage is associated with sperm DNA damage in these patients. PMID- 16361287 TI - Meiotic segregation patterns and ICSI pregnancy outcome of a rare (13;21) Robertsonian translocation carrier: a case report. AB - t(13;21) is an uncommon Robertsonian translocation (RT) with limited information in the literature. Hereby, we assessed the meiotic segregation and interchromosomal effect (ICE) in sperm nuclei from a t(13;21) carrier. The pregnancy outcome following ICSI was also included as reference for physicians and patients. Dual-colour fluorescent in situ hybridization (FISH) was carried out to analyse the segregation pattern of chromosomes 13 and 21, while triple colour FISH was used to investigate the possible concurrence of ICE. With respect to chromosomal constitutions of 13 and 21, 88.39% of the spermatozoa were normal or balanced due to alternative segregations, and 11.08% showed nullisomy or disomy as a result of adjacent segregations. However, for chromosome 18 and sex chromosomes, the proportion of normal haploids was 98.79%. The rate of disomy was not significantly higher than the controls for either chromosome 18 or X/Y. The rare t(13;21) case exhibited a similar pattern of meiotic segregation as in the common RTs. ICEs were not observed in the current case. PMID- 16361288 TI - Influence of analytical and biological variation on the clinical interpretation of seminal parameters. AB - Quality assurance in semen analysis has been questioned recently in this journal. Based on the limited capacity of seminal parameter in the determination of fertility, the authors advocated abandoning methods of quality assurance in semen analysis for clinical situations. In this article, we explore arguments as to why quality assurance in semen analysis for clinical use is not 'a waste of time'. Imprecision and within-subject biological variations are the two major components involved in the dispersion of seminal parameter results obtained by analysis of a semen sample from an individual. As within-subject biological variation is constant across geography, time and population, imprecision is a very important factor in the quality of laboratory test results. We analyse this influence on various seminal parameters and observe that there is an amount of error that can be tolerated without invalidating the medical usefulness of seminal parameter determination. However, there is a maximum allowable analytical error above which the medical usefulness of seminal parameter results is invalidated. The level of performance required to facilitate clinical decision-making is termed quality specification. We comment on different strategies to define the maximum allowable analytical error. PMID- 16361289 TI - The association between polycystic ovaries and endometrial cancer. AB - BACKGROUND: Women with polycystic ovary syndrome (PCOS) are assumed to be at increased risk of endometrial cancer (EC), albeit of a more differentiated type with better prognosis than in normal women. This study was designed to test these assumptions, as evidence for them is lacking. METHODS: The prevalence of polycystic ovaries (PCO), as a marker of PCOS, was investigated in ovarian sections from 128 women with EC and 83 with benign gynaecological conditions. The expression of the prognostic markers p53, Ki67, Bcl2 and cyclin D1 was also investigated by immunohistochemistry in endometrial tumours from 11 women with PCO and 16 with normal ovaries. RESULTS: Overall, PCO were similarly prevalent in women with EC (8.6%) and benign controls (8.4%); however, in women aged <50 years, PCO were more prevalent in women with EC (62.5 versus 27.3%, P = 0.033). Cyclin D1-expressing endometrial tumours tended to be more prevalent in women with PCO compared to normal ovaries (36.4 versus 6.25%, respectively, P = 0.071). Bcl2-, p53- and Ki67-expressing tumours were similarly prevalent. CONCLUSIONS: The association between PCOS and EC appears confined to premenopausal women. The tendency for cyclin D1-expressing endometrial tumours to be more prevalent in women with PCO challenges the assumption that EC prognosis is improved in women with PCOS. PMID- 16361290 TI - ICSI outcomes in obstructive azoospermia: influence of the origin of surgically retrieved spermatozoa and the cause of obstruction. AB - BACKGROUND: Spermatozoa can be retrieved from the testis and epididymis of men with obstructive azoospermia (OA) and used for ICSI. However, it is unknown whether the outcome of ICSI depends on the cause of obstruction or the origin of surgically retrieved spermatozoa. METHODS: A cohort of 171 men with OA and normal spermatogenesis were included in this retrospective study. They were divided into three groups according to the site and origin of obstruction: 83 men had congenital bilateral absence of vas deferens; 55 and 33 had acquired epididymal and deferent duct obstructions, respectively. The outcome of 368 ICSI cycles was determined and compared according to the origin of spermatozoa: epididymal (n = 253) or testicular (n = 115). RESULTS: Fertilization and clinical pregnancy rates did not differ between spermatozoa of different origin (58.9% versus 51.9% and 22.1% versus 24.3% with epididymal and testicular spermatozoa, respectively). However, the miscarriage rate was significantly higher for testicular spermatozoa (35.7% versus. 12.5% P < 0.05, chi2 test). Findings were similar whatever the aetiology of the OA. CONCLUSION: This study suggests that the use of testicular spermatozoa, even those generated during normal spermatogenesis, alters embryonic development and that epididymal spermatozoa should be preferentially used, irrespective of the aetiology of OA. PMID- 16361291 TI - Pituitary-ovarian axis during lactational amenorrhoea. II. Longitudinal assessment of serum FSH polymorphism before and after recovery of menstrual cycles. AB - BACKGROUND: The association of normal serum levels of immunoassayable gonadotrophins with anovulation during lactational amenorrhoea (LA) has not been fully explained. METHODS: Serum FSH polymorphism was analysed in 10 women during LA between days 60 and 70 post-partum and again, in the mid-follicular phase (MFP), after resuming menstrual cyclicity. FSH microheterogeneity was characterized according to charge, using preparative isoelectric focusing, and according to the inner structure of carbohydrate chains, using lectin chromatography. RESULTS: A significantly higher proportion of FSH charge isoforms isolated below pH 4.10 and a lower proportion of FSH isoforms bearing highly branched oligosaccharides were observed during LA when compared to MFP. Further analysis with higher resolution showed that FSH charge isoforms, isolated in the lower pH range in LA, corresponded to FSH molecules bearing highly branched and biantennary oligosaccharides. FSH isoforms bearing hybrid-type oligosaccharides were only present during LA. The circulating FSH isoform mix was significantly less bioactive in LA than in MFP. LA is characterized by a more acidic mix of FSH isoforms, containing hormone bearing less processed oligosaccharides, with decreased biopotency in comparison with the follicular phase. CONCLUSIONS: This FSH microheterogeneity may be one of the critical factors contributing to incomplete follicular development and anovulation during LA. PMID- 16361292 TI - Pituitary-ovarian axis during lactational amenorrhoea. I. Longitudinal assessment of follicular growth, gonadotrophins, sex steroids and inhibin levels before and after recovery of menstrual cyclicity. AB - BACKGROUND: Comparisons of follicular development and hormonal profile in the same women during and after lactational amenorrhoea (LA) are scarce. We report follicular growth, pituitary and ovarian hormone serum levels in the same women during LA and in follicular phases after resumption of menstrual cyclicity. METHODS: Serum samples were obtained from 10 women during LA between days 60 and 89 post-partum and between days 1 and 4 (early follicular phase; EFP) and 7 and 10 (mid-follicular phase; MFP) of the second and third cycles after LA. RESULTS: The number of follicles >3 mm and diameter of the largest follicle were significantly higher during LA when compared to EFP and MFP. Serum levels of inhibin B were similar in LA and EFP and increased significantly in MFP. Pro alphaC was significantly higher in EFP than in LA and MFP. Estradiol was similar during all stages. In comparison with EFP and MFP, LA is associated with higher prolactin levels, normal or slightly elevated gonadotrophins and increased number and size of follicles without a parallel increase in estradiol, inhibin B and Pro alphaC. CONCLUSIONS: During LA, there is a profound dissociation between follicular growth and follicular endocrine activity, which suggests an alteration in the stimulus-response relationship at the follicular level. PMID- 16361293 TI - Double trisomy in spontaneous miscarriages: cytogenetic and molecular approach. AB - BACKGROUND: Although single trisomy is the most common chromosomal abnormality observed within first trimester spontaneous abortions (SA) (>50%), double trisomy (DT) ranges from 0.21 to 2.8% in the literature. Since little is known about mechanisms underlying DT, we report the results of our experience with 517 SA, establishing parental origin and cell stage of non-disjunction when possible in DT cases, and making a revision of those previously reported. METHODS: Cytogenetic analysis was performed in all aborted specimens. Quantitative fluorescent PCR (QF-PCR) and multiplex ligation-dependent probe amplification (MLPA) were performed in DT cases in order to assess parental origin and stage of error of aneuploidy in addition to its reliability in detecting aneuploidies. RESULTS: Karyotyping was successful in 321 miscarriages; the rate of DT was 2.18%. Among the seven DT cases reported, three new combinations were found. Maternal origin was established for all DT SA analysed. Meiotic stage of error was presumed meiosis I (MI) for 48,XX+15+22 and 48,XX+8+21, meiosis II (MII) for 48,XXX+18, and MII and MI respectively for 48,XY+18+22. Molecular results agreed with cytogenetic results. CONCLUSIONS: Similar maternal age-related mechanisms could be implicated in both single and double trisomy. Molecular techniques could be useful in diagnosing not only single but multiple aneuploidy and determining its origin. This will improve our knowledge about mechanisms underlying human aneuploidy, and enable appropriate genetic counselling. PMID- 16361294 TI - Assisted reproductive therapies and imprinting disorders--a preliminary British survey. AB - BACKGROUND: Recent reports have suggested a higher risk of Beckwith-Wiedemann syndrome (BWS) and Angelman syndrome (AS) after assisted reproductive technologies (ARTs), but it is unclear whether this might also apply to other disorders of genomic imprinting. METHODS: We contacted families of children with BWS, AS, Prader-Willi syndrome (PWS) and transient neonatal diabetes mellitus (TNDM) to determine use of ART. RESULTS: A statistically significant increased frequency of ART in children with BWS was confirmed [2.9%, 95% confidence interval (CI) 1.4-6.3% vs 0.8% expected] but there was no significant association with PWS or TNDM. Consideration of the molecular subgroup of BWS and AS suggested the feasibility of association with ART. CONCLUSIONS: These differences may relate to variations in (i) the molecular mechanisms for disordered imprinting in the different disorders and (ii) the susceptibility of specific imprinting control regions to ART-associated methylation alterations (epimutations). PMID- 16361295 TI - Time to pregnancy as a function of male and female serum concentrations of 2,2'4,4'5,5'-hexachlorobiphenyl (CB-153) and 1,1-dichloro-2,2-bis (p chlorophenyl)-ethylene (p,p'-DDE). AB - BACKGROUND: Persistent organochlorine pollutants (POP) may affect both the female and male reproductive system in animals as well as in humans. METHODS: Blood samples were collected from pregnant women and their partners from Greenland, Warsaw and Kharkiv, and from a cohort of Swedish fishermen's wives. Blood samples were analysed for 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153) and 1,1-dichloro-2,2 bis (p-chlorophenyl)-ethylene (p,p'-DDE). Information on the participants' fertility, measured as time to pregnancy (TTP), was collected. In total, 778 men and 1505 women were included in the analyses. RESULTS: The data from Warsaw, Kharkiv and the Swedish fishermen's wives indicated no effect of either male or female exposure to POP on TTP. However, among men and women from Greenland, there seemed to be an association between serum concentrations of CB-153 and p,p'-DDE and prolonged TTP. Due to the strong intra-individual correlation between CB-153 and p,p'-DDE in the Greenlandic population, it was not possible to determine whether the risk was associated with CB-153 or p, p'-DDE or was an interaction between the two compounds. CONCLUSIONS: The overall results of the present study create a somewhat ambiguous pattern, but give some support to the idea that dietary POP exposure might be harmful for couple fertility. PMID- 16361297 TI - Use of transoesophageal echocardiography during cardiac arrest in patients undergoing elective non-cardiac surgery. AB - BACKGROUND: Sudden unexpected intraoperative cardiac arrests in patients undergoing elective non-cardiac operations are rare but catastrophic complications. The efficacy and utility of transoesophageal echocardiography (TOE) in the diagnosis and management of these events have not been reported earlier. METHODS: Unexpected intraoperative cardiac arrests in patients undergoing elective non-cardiac operations were prospectively studied during a 6 yr period. In these patients, TOE was performed within 5 min after initiation of external chest compression to identify the causes of the cardiac arrests. RESULTS: Among a total of 125 965 surgical operations, 10 patients who suffered intraoperative cardiac arrests were studied. The causes of the cardiac arrests including myocardial infarction in five, pulmonary embolism in two, and severe hypovolaemia and ventricular arrhythmia without specific pathology in the other two patients were correctly identified using TOE. Seven patients survived. CONCLUSION: We conclude that TOE was useful in cardiac arrest during non-cardiac surgery for identifying or excluding pulmonary embolism and for directing investigations and treatment in those patients who had suffered myocardial infarction. PMID- 16361296 TI - Treatment with the GnRH antagonist ganirelix prevents premature LH rises and luteinization in stimulated intrauterine insemination: results of a double-blind, placebo-controlled, multicentre trial. AB - BACKGROUND: This study was designed to assess whether the use of ganirelix in women undergoing stimulated IUI could prevent the occurrence of premature LH rises and luteinization (LH+progesterone rises). METHODS: Women of infertile couples, diagnosed with unexplained or male factor infertility, were randomized to receive either ganirelix (n=103) or placebo (n=100) in a double-blind design. All women were treated with an individualized, low-dose rFSH regimen started on day 2-3 of cycle. Ganirelix (0.25 mg/day) was started if one or more follicles>or=14 mm were visualized. Ovulation was triggered by HCG injection when at least one follicle>or=18 mm was observed and a single IUI was performed 34-42 h later. The primary efficacy outcome was the incidence of premature LH rises (+/ progesterone rise). RESULTS: In the ganirelix group, four subjects had a premature LH rise (value>or=10 IU/l), one LH rise prior to the start of ganirelix and three LH rises during ganirelix treatment, whereas in the placebo group 28 subjects had a premature LH rise, six subjects prior to the start of placebo and 22 subjects during placebo treatment. The incidence of LH rises was significantly lower in ganirelix cycles compared to placebo cycles (3.9 versus 28.0%; P=0.003 for ITT analysis). When excluding subjects with an LH value>or=10 IU/l before the start of ganirelix/placebo the incidence of LH rises was also significantly lower in ganirelix cycles compared to placebo cycles (2.9 versus 23.4%; P=0.003 for ITT analysis). Premature luteinization (LH rise with concomitant progesterone rise>or=1 ng/ml) was observed in one subject in the ganirelix group and in 17 subjects in the placebo group of which three subjects had a premature spontaneous ovulation. Ongoing pregnancy rates per attempt were 12.6 and 12.0% for the ganirelix and placebo groups respectively. CONCLUSIONS: Treatment with ganirelix effectively prevents premature LH rises, luteinization in subjects undergoing stimulated IUI. Low-dose rFSH regimen combined with a GnRH antagonist may be an alternative treatment option for subjects with previous proven luteinization or in subjects who would otherwise require insemination when staff are not working. PMID- 16361298 TI - Reversible nitrous oxide myelopathy and a polymorphism in the gene encoding 5,10 methylenetetrahydrofolate reductase. AB - We present a case of a patient who received nitrous oxide on two occasions within a period of 8 weeks and who subsequently developed a diffuse myelopathy, characterized by upper extremity paresis, lower extremity paraplegia and neurogenic bladder. Laboratory testing revealed hyperhomocysteinaemia and low levels of vitamin B(12). Because of this uncommon clinical presentation, we analysed the patient's DNA, and found a polymorphism in the MTHFR gene that is associated with the thermolabile isoform of the 5,10-methylenetetrahydrofolate reductase enzyme, which explained the myelopathy experienced by the patient after being exposed to nitrous oxide. Soon after initiating supplementary therapy with folic acid and vitamin B(12), the neurological symptoms subsided. PMID- 16361299 TI - Alveolar recruitment in acute lung injury. AB - Alveolar recruitment is one of the primary goals of respiratory care for acute lung injury. It is aimed at improving pulmonary gas exchange and, even more important, at protecting the lungs from ventilator-induced trauma. This review addresses the concept of alveolar recruitment for lung protection in acute lung injury. It provides reasons for why atelectasis and atelectrauma should be avoided; it analyses current and future approaches on how to achieve and preserve alveolar recruitment; and it discusses the possibilities of detecting alveolar recruitment and derecruitment. The latter is of particular clinical relevance because interventions aimed at lung recruitment are often undertaken without simultaneous verification of their effectiveness. PMID- 16361300 TI - Single dose parecoxib significantly improves ventilatory function in early extubation coronary artery bypass surgery: a prospective randomized double blind placebo controlled trial. AB - BACKGROUND: Parecoxib, a cyclo oxygenase-2 inhibiting non-steroidal anti inflammatory drug, has been widely used for postoperative analgesia. Our aim was to quantify the benefit of a single dose after coronary artery bypass grafting. METHODS: The investigation was carried out as a randomized double blind placebo controlled study. A single i.v. dose of parecoxib 40 mg or placebo was given at closure of sternotomy. No opioid other than morphine was given in the first 24 postoperative hours. Pain was assessed using both a Visual Analogue Score (1-10), and the amount of morphine used via a morphine patient controlled analgesia pump. Creatinine clearance was measured before and after operation from 24 h urine collections. After a global announcement by Pfizer that paracoxib was 'contraindicated in patients with ischaemic heart disease' further recruitment was suspended and the collected data from 40 patients were analysed. RESULTS: Twenty-one patients received parecoxib and 19 received placebo. Amongst those who received parecoxib, there was a highly significant sparing of rescue medication before tracheal extubation (P=0.004) compared with placebo, and an overall 35% morphine sparing effect during the first 6 h post extubation after correction for the variability in extubation time (P=0.037). Respiration, as measured by arterial carbon dioxide tension at the time of extubation, was significantly better in the parecoxib group (P=0.045). Significantly more furosemide was given for postoperative oliguria in those patients who received parecoxib (P=0.036). After correcting for differences in diuretic usage and fluid balance, parecoxib was associated with a significant increase in plasma creatinine (P=0.041). CONCLUSION: A single dose of parecoxib has a significant opioid sparing effect in the first 6 h after coronary artery bypass grafting which resulted in significantly improved ventilation with mild elevation of plasma creatinine within normal limits. PMID- 16361301 TI - Epidural haematoma after a combined spinal-epidural anaesthetic in a patient treated with clopidogrel and dalteparin. AB - We report a case of a spinal-epidural haematoma occurring in a patient after a combined spinal-epidural anaesthetic. She had been taking clopidogrel and had received perioperative dalteparin for thromboprophylaxis. Despite adhering to standard guidelines concerning administration of low molecular weight heparin perioperatively and stopping the clopidogrel 7 days before the anaesthetic, the patient developed an epidural haematoma. PMID- 16361302 TI - Effect of oral gabapentin on postoperative epidural analgesia. AB - BACKGROUND: Gabapentin has been used successfully as a non-opioid analgesic adjuvant for postoperative pain management. We hypothesized that gabapentin might be a useful adjuvant for postoperative analgesia provided with patient-controlled epidural analgesia (PCEA). METHODS: Forty patients undergoing lower extremity surgery procedures were randomly assigned to receive (i) placebo capsules (control) or (ii) gabapentin (1.2 g day(-1)) before and for 2 days after surgery. Anaesthetic technique was standardized. Postoperative assessments included verbal rating scale scoring for pain and sedation, PCEA usage, quality of recovery assessment, times of GI function recovery, and patient satisfaction scoring for pain management. RESULTS: Pain scores at 1, 4, 8, 12, and 16 h (P<0.001), PCEA bolus requirements (n) at 24 [21 (3), 14 (2)], 48 [15 (4), 10 (3)] and 72 [8 (5), 2 (3)] (P<0.05) and paracetamol (mg) consumption [700 (523), 350 (400)]; P<0.05), were significantly lower in the gabapentin-treated patients than in the control group. Patient satisfaction with postoperative pain management at 24 h was better in gabapentin-treated patients [85.5 (7.5), 66.5 (15)]; P<0.001). Gabapentin treated patients had less motor block when compared with control group. Times of return of bowel function, hospitalization, and resumption of dietary intake were similar in the groups. However, the incidence of dizziness was higher in the gabapentin group (35% vs 5%; P<0.05). CONCLUSIONS: Oral gabapentin (1.2 g day( 1)) as an adjunct to epidural analgesia decreased pain and analgesic consumption. Despite an increased incidence of dizziness it also increased patient satisfaction. PMID- 16361303 TI - The dazzling array of basal branches in the mtDNA macrohaplogroup M from India as inferred from complete genomes. AB - Many efforts based on complete mitochondrial DNA (mtDNA) genomes have been made to depict the global mtDNA landscape, but the phylogeny of Indian macrohaplogroup M has not yet been resolved in detail. To fill this lacuna, we took the same strategy as in our recent analysis of Indian mtDNA macrohaplogroup N and selected 56 mtDNAs from over 1,200 samples across India for complete sequencing, with the intention to cover all Indian autochthonous M lineages. As a result, the phylogenetic status of previously identified haplogroups based on control-region and/or partial coding-region information, such as M2, M3, M4, M5, M6, M30, and M33, was solidified or redefined here. Moreover, seven novel basal M haplogroups (viz., M34-M40) were identified, and yet another five singular branches of the M phylogeny were discovered in the present study. The comparison of matrilineal components among India, East Asia, Southeast Asia, and Oceania at the deepest level yielded a star-like and nonoverlapping pattern, reflecting a rapid mode of modern human dispersal along the Asian coast after the initial "out-of-Africa" event. PMID- 16361304 TI - Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Molecular genetics of natural populations. AB - Significant progress in evolutionary genetics has been made by studying, on the one hand, patterns of DNA sequence polymorphism and, on the other, genetic architecture of complex adaptive traits. However, connections between nucleotide variants under selection and adaptively relevant phenotypes are missing. Such connections can be established using precise gene replacement. We review the recent successful introduction of this technique to the analysis of two evolutionarily interesting loci--Odysseus and desaturase2. Both genes have subtle phenotypes that nevertheless could be identified using gene replacement, demonstrating that effects of naturally occurring alleles can be measured in the laboratory. This is an important first step in connecting statistical signatures of selection with adaptation in nature. More candidate genes involved in adaptation, for example, through cloning of genes responsible for reproductive isolation, now need to be identified. Molecular genetic manipulation, DNA polymorphism analysis, and field studies then have to be integrated to provide fresh insights into the mechanisms of evolutionary change. PMID- 16361305 TI - Evidence for association of OCTN genes and IBD5 with ulcerative colitis. AB - BACKGROUND AND AIMS: Genetic association between Crohn's disease (CD) and OCTN1 (SLC22A4) C1672T/OCTN2 (SLC22A5) G-207C variants in IBD5 has recently been reported. These genes encode solute carriers and the association was suggested to be distinct from the background IBD5 risk haplotype. There have been conflicting reports of the association between markers in the IBD5 region and ulcerative colitis (UC) and interaction (epistasis) between this locus and CARD15. Our aim was to ascertain the contribution of OCTN variants to UC and CD in a large independent UK dataset, to seek genetic evidence that the OCTN association is distinct from the IBD5 risk haplotype and to identify interactions between the IBD5 and CARD15 loci. METHODS: A total of 1104 unrelated Caucasian subjects with inflammatory bowel disease (IBD) (496 CD, 512 UC, 96 indeterminate) and 750 ethnically matched controls were genotyped for three single nucleotide polymorphisms (SNPs) in the CD associated genes (OCTN1+1672, OCTN2-207, and IGR2230), and two flanking IBD5 tagging SNPs, IGR2096 and IGR3096. Data were analysed by logistic regression methods within STATA. RESULTS: OCTN variants were as strongly associated with UC and IBD overall as they were with CD (p = 0.0001; OR 1.3 (95% confidence interval 1.1-1.5)). OCTN variants were in tight linkage disequilibrium with the extended IBD5 risk haplotype D' 0.79 and 0.88, and r2 = 0.62 and 0.72 for IGR2096 and 3096, respectively. There was no deviation from a multiplicative model of interaction between CARD15 and IBD5 on the penetrance scale. CONCLUSIONS: The OCTN variants were associated with susceptibility to IBD overall. The effect was equally strong in UC and CD. Although OCTN variants may account for the increased risk of IBD associated with IBD5, a role for other candidate genes within this extended haplotype was not excluded. There was no statistical evidence of interaction between CARD15 and either OCTN or IBD5 variants in susceptibility to IBD. PMID- 16361306 TI - Phenotype at diagnosis predicts recurrence rates in Crohn's disease. AB - BACKGROUND: In Crohn's disease (CD), studies associating phenotype at diagnosis and subsequent disease activity are important for patient counselling and health care planning. AIMS: To calculate disease recurrence rates and to correlate these with phenotypic traits at diagnosis. METHODS: A prospectively assembled uniformly diagnosed European population based inception cohort of CD patients was classified according to the Vienna classification for disease phenotype at diagnosis. Surgical and non-surgical recurrence rates throughout a 10 year follow up period were calculated. Multivariate analysis was performed to classify risk factors present at diagnosis for recurrent disease. RESULTS: A total of 358 were classified for phenotype at diagnosis, of whom 262 (73.2%) had a first recurrence and 113 patients (31.6%) a first surgical recurrence during the first 10 years after diagnosis. Patients with upper gastrointestinal disease at diagnosis had an excess risk of recurrence (hazard ratio 1.54 (95% confidence interval (CI) 1.13 2.10)) whereas age >/=40 years at diagnosis was protective (hazard ratio 0.82 (95% CI 0.70-0.97)). Colonic disease was a protective characteristic for resective surgery (hazard ratio 0.38 (95% CI 0.21-0.69)). More frequent resective surgical recurrences were reported from Copenhagen (hazard ratio 3.23 (95% CI 1.32-7.89)). CONCLUSIONS: A mild course of disease in terms of disease recurrence was observed in this European cohort. Phenotype at diagnosis had predictive value for disease recurrence with upper gastrointestinal disease being the most important positive predictor. A phenotypic North-South gradient in CD may be present, illustrated by higher surgery risks in some of the Northern European centres. PMID- 16361307 TI - A novel integrin alpha5beta1 binding domain in module 4 of connective tissue growth factor (CCN2/CTGF) promotes adhesion and migration of activated pancreatic stellate cells. AB - BACKGROUND: Connective tissue growth factor (CCN2) is upregulated in pancreatic fibrosis and desmoplastic pancreatic tumours. CCN2 interacts with integrin alpha5beta1 on pancreatic stellate cells (PSC) in which it stimulates fibrogenesis, adhesion, migration, and proliferation. AIM: To determine the structural domain(s) in CCN2 that interact with integrin alpha5beta1 to regulation PSC functions. METHODS: Primary activated rat PSC were tested for their adherence to isoforms of CCN2 comprising modules 1-4 (CCN2(1-4)), modules 3 4 (CCN2(3-4)), module 3 alone (CCN2(3)), or module 4 alone (CCN2(4)). Adhesion studies were performed in the presence of EDTA, divalent cations, anti-integrin alpha5beta1 antibodies, CCN2 synthetic peptides, or heparin, or after pretreatment of the cells with heparinase, chondroitinase, or sodium chlorate. CCN2 integrin alpha5beta1 binding was analysed in cell free systems. The ability of CCN2(1-4), CCN2(3-4), or CCN2(4) to stimulate PSC migration was evaluated in the presence of anti-integrin alpha5beta1 or heparin. RESULTS: PSC adhesion was stimulated by CCN2(1-4), CCN2(3-4), or CCN2(4) and supported by Mg2+ but not Ca2+. CCN2(4) supported PSC adhesion or migration were blocked by anti-integrin alpha5beta1 antibodies or by treatment of cells with heparinase or sodium chlorate. A direct interaction between CCN2(4) and integrin alpha5beta1 was demonstrated in cell free assays. The sequence GVCTDGR in module 4 mediated the binding between CCN2(4) and integrin alpha5beta1 as well as CCN2(4) mediated PSC adhesion and migration. CONCLUSIONS: A GVCTDGR sequence in module 4 of CCN2 is a novel integrin alpha5beta1 binding site that is essential for CCN2 stimulated functions in PSC and which represents a new therapeutic target in PSC mediated fibrogenesis. PMID- 16361308 TI - Positron emission tomography scanning is not superior to whole body multidetector helical computed tomography in the preoperative staging of colorectal cancer. AB - BACKGROUND: The role of positron emission tomography with the glucose analogue [18F] fluoro-2-deoxy-D-glucose (FDG-PET) in the initial staging of disease in patients with primary colorectal cancer (CRC) has not been adequately assessed. AIMS: To evaluate the additional value of FDG-PET as a staging modality, complementary to routine multidetector row computed tomography (MDCT) in patients with CRC. METHODS: Forty four patients with CRC underwent preoperative MDCT and FDG-PET. The accuracy of intraoperative macroscopic staging was also investigated compared with histopathological diagnosis. All FDG-PET images were evaluated with respect to detectability of the primary tumour, lymph node involvement, and distant metastases. Both MDCT and FDG-PET diagnoses and treatment plan were compared with surgical and histopathological results. RESULTS: Thirty seven patients underwent surgery. Tumour detection rate was 95% (42/44) for MDCT, 100% (44/44) for FDG-PET, and 100% (37/37) for intraoperative macroscopic diagnosis. Pathological diagnosis of T factor was T1 in five, T2 in four, T3 in 24, and T4 in four cases. Concordance rate with pathological findings of T factor was 57% (21/37) for MDCT and 62% (23/37) for macroscopic diagnosis. Lymph node involvement was pathologically positive in 19 cases. Regarding N factor, overall accuracy was 62% (23/37) for MDCT, 59% (22/37) for FDG-PET, and 70% (26/37) for macroscopic diagnosis. For all 44 patients, FDG-PET findings resulted in treatment changes in only one (2%) patient. CONCLUSION: FDG-PET is not superior to routine MDCT in the initial staging of primary CRC. PMID- 16361309 TI - Inhibitory effects of Lactobacillus reuteri on visceral pain induced by colorectal distension in Sprague-Dawley rats. AB - BACKGROUND AND AIMS: Probiotic bacteria are being investigated as possible treatments for many intestinal disorders. The present study aimed to explore the effects of live, heat killed, or gamma irradiated Lactobacillus reuteri on cardio autonomic response and single fibre unit discharge in dorsal root ganglia to colorectal distension in healthy Sprague-Dawley rats housed under conventional conditions. The effects of this treatment on somatic pain were also examined. METHODS: 1x10(9) bacteria were given by gavage for nine days. Colorectal distension occurred under anaesthesia. Heart rate was measured through continuous electrocardiography. Single fibre unit discharge was recorded from the 6th left lumbar dorsal root ganglion. Somatic pain was evaluated by the tail flick and paw pressure tests. RESULTS: Colorectal distension caused a pressure dependent bradycardia in the control (native medium) group. Treatment with live, heat killed, or gamma irradiated bacteria as well as their products (conditioned medium) prevented the pain response even during the maximum distension pressure (80 mm Hg). Both viable and non-viable bacteria significantly decreased dorsal root ganglion single unit activity to distension. No effects on somatic pain were seen with any treatment. CONCLUSIONS: Oral administration of either live or killed probiotic bacteria or conditioned medium inhibited the constitutive cardio autonomic response to colorectal distension in rats through effects on enteric nerves. These data may provide a novel explanation for beneficial probiotic effects on visceral pain. PMID- 16361310 TI - Targeting CD45RB alters T cell migration and delays viral clearance. AB - CD45 is a receptor tyrosine phosphatase essential for TCR signaling. One isoform, CD45RB, is down-regulated in memory cells and targeting CD45RB with a specific antibody has been shown to inhibit graft rejection. Its role in immunity to infection, however, has not been tested. Here, we report the effect of anti CD45RB antibody treatment on the induction of anti-influenza CD8+ T cells and viral clearance. Anti-CD45RB-treated mice had delayed pulmonary viral clearance compared with untreated mice whose infection was completely cleared by day 8 post infection. In anti-CD45RB-treated mice, the total CD4+ and CD8+ T cell numbers in both the lungs and mediastinal nodes were substantially reduced at days 5 and 8; this effect was less marked for the spleen. CD8+ T cells specific for influenza virus were also reduced compared with the control group in all three organs at day 8. By day 11, when both treated and control groups showed no virus remaining in the lungs, specific CD8+ T cell numbers were at similar low levels. Homing to lymph nodes and lung of dye-labeled T cells was greatly inhibited (by >80%) by anti-CD45RB treatment. This reduced homing corresponded with reduced CD62L and beta1-integrin expression in both uninfected and infected mice. Since CD62L plays a critical role in homing lymphocytes to lymph nodes, and high levels of CD62L and alpha4beta1-integrin are expressed by lymphocytes that home to bronchus associated lymphoid tissue, we suggest that reduced expression of these molecules is a key explanation for the delay in immune responses. PMID- 16361311 TI - Regulatory T cell-like activity of Foxp3+ adult T cell leukemia cells. AB - Adult T cell leukemia (ATL) is an aggressive neoplastic disease, in which a quarter of the patients develop opportunistic infections due to cellular immunodeficiency. However, the underlying mechanism responsible for the immunosuppression has remained unclear. Recent studies have demonstrated that the leukemia cells from a subset of patients with ATL express Foxp3, a specific marker for CD25+CD4+ regulatory T (Treg) cells, which regulate the immune response by suppressing CD4+ T cell functions. However, whether there is a functional resemblance between ATL cells that have Foxp3 expression and Treg cells is still unknown. In this report, we confirmed the high expression of Foxp3 in leukemia cells from 5 of 12 ATL patients and demonstrated that ATL cells from 3 patients suppressed the proliferation of CD4+ T cells. Similarly, one of six HTLV-I-infected cell lines showed both high Foxp3 expression and suppressive activity. Like Treg cells, the suppression induced by the ATL cells from two patients and the HTLV-infected cell line appeared to be mediated by a cell-cell contact-dependent mechanism. Nevertheless, among the ATL cells that strongly expressed Foxp3, those from two of the five patients showed no apparent suppressive activity. Furthermore, retroviral transfection of Foxp3 did not confer any suppressive function on low Foxp3-expressing HTLV-I-infected cell lines. These results indicate that Foxp3 may be essential but is not sufficient for the Treg-cell-like suppressive activity of ATL cells and HTLV-I-infected cell lines. PMID- 16361312 TI - The amino acid at position 97 is involved in folding and surface expression of HLA-B27. AB - HLA-B27 confers susceptibility to ankylosing spondylitis (AS) but the mechanism linking this association remains unknown. Other properties unrelated to its natural role of antigen presenting function may be important in disease pathogenesis. We determined here the impact of N97D substitution on the folding and expression of HLA-B*2704 transfected in the 721.221 cell line. The mutation at position 97 abolishes the surface expression of non-conformational (HC10) and conformational (ME1) forms. The expression of ME1 forms was found to be absent in B*2704 N97D by immunoprecipitation and flow cytometry of fixed and permeabilized cell experiments with the conformation-sensitive ME1 antibody. However, immunoblotting cell lysates with HC10 revealed the presence of unfolded heavy chain (HC) and HC-dimer forms. The impact of the N97D mutation in the exit from the endoplasmic reticulum (ER) was analysed by western blot after endoglycosidase H treatment, and it was found that B*2704 N97D was retained and accumulated as unfolded molecules. We tested for mutant association with transporter associated with antigen processing (TAP), calnexin (CNX), calreticulin (CLR) and beta2 microglobulin (beta2m). The wild-type B*2704 and N97D mutants were associated with TAP, CNX and CLR, although HC10 forms of mutant N97D interact more weakly with TAP. Only folded molecules of HLA-B*2704 were associated with beta2m. Surprisingly, the peptide-binding assay demonstrated the ability of unfolded N97D molecules to bind high-affinity peptides. It has been suggested that AS may arise because of aberrant folding of HLA-B27 molecules within the ER. Future work must therefore aim to clarify the functional connection between the unfolded protein response pathway in response to the accumulation of HLA-B27 in the ER. This mutant could be useful as a model for the misfolding of HLA-B27. PMID- 16361313 TI - Presentation of self-antigens on MHC class II molecules during dendritic cell maturation. AB - Little is known about how dendritic cells (DCs) maintain a balance between tolerance and immunity for antigens synthesized by DCs themselves. Using transgenic DCs expressing a model self-antigen, in vitro self-peptide-MHC class II complex formation and presentation increased with DC maturation, as for exogenous antigens. In vivo, however, even 'immature' DCs isolated from steady state lymph nodes expressed MHC at mature cell levels, although many were also CD86 low. Adoptive transfer of naive specific T cells into unstimulated transgenic mice resulted in tolerance. If the mice were also injected with anti CD40 or Listeria monocytogenes, there was robust specific T cell expansion and inflammation. Thus, DC-endogenous antigens may induce tolerance, but only in the absence of potent maturation stimuli. PMID- 16361314 TI - Subtle sequence variation among MHC class I locus products greatly influences sensitivity to HCMV US2- and US11-mediated degradation. AB - Human cytomegalovirus (HCMV) interferes with cellular immune responses by modulating surface expression of MHC class I molecules. Here, we focused on HCMV encoded unique short (US) 2 and US11, which bind newly synthesized MHC class I heavy chains (HCs) and support their dislocation into the cytosol for subsequent degradation by proteasomes. Not all MHC class I locus products are equally sensitive to this down-modulation. The aim of this study was to identify which domains, and ultimately which residues, are responsible for the resistance or sensitivity of MHC class I molecules to US2- and US11-mediated down-regulation. We show that, besides endoplasmic reticulum-lumenal regions, the C-terminus of class I molecules represents an important determinant for allele specificity in US11-mediated degradation. HLA-E becomes sensitive to US11-mediated down regulation when its cytoplasmic tail is extended. Interestingly, this only requires two additional residues, lysine and valine, at its C-terminus. For US2, the MHC class I allele specificity is largely determined by a small region at the junction of the alpha2/alpha3 domain of the HC. It is quite remarkable that minor changes, in only four residues, can completely revert the sensitivity of naturally US2-resistant HLA-E molecules. With this study we provide better insights into the features underlying the selectivity in MHC class I down regulation by US2 and US11. PMID- 16361315 TI - Only VpreB1, but not VpreB2, is expressed at levels which allow normal development of B cells. AB - The surrogate light chain (SLC) consists of the polypeptides lambda5 and, in the mouse, either VpreB1 or VpreB2. SLC associates with BILL-Cadherin and other glycoproteins to form the pro-B cell receptor (pro-BCR) at the pre-BI cell stage, and with the immunoglobulin mu heavy chain to form the pre-BCR at the pre-BII cell stage. The function of the pro-BCR, if any, is unknown, whereas the pre-BCR is crucial for proliferative expansion of pre-BII cells. To shed light on the functional properties of VpreB1 and VpreB2 in vivo, mice with either one or two VpreB1, or one or two VpreB2, alleles have been investigated. We show that B cell development in mice with two VpreB1 alleles is indistinguishable from that of normal mice. In contrast, mice with two VpreB2 alleles show an approximately 1.6 fold increase in pre-BI and a 35% decrease in pre-BII cell numbers, while mice with only one VpreB2 allele show a reduction in B cell development manifested in a 2-fold enrichment in pre-BI cells and a 75% reduction in pre-BII cells. However, such a gene dosage effect is not observed for VpreB1. Our results suggest that the difference between VpreB1- and VpreB2-deficient mice is due to lower VpreB2 protein expression, thus limiting the formation of pre-BCRs and thereby the number of large, cycling pre-BII cells. PMID- 16361316 TI - Experimental autoimmune encephalomyelitis in mice expressing the autoantigen MBP 1-10 covalently bound to the MHC class II molecule I-Au. AB - Most autoantigens implicated in multiple sclerosis (MS) are expressed not only in the central nervous system (CNS) but also in the thymus and the periphery. Nevertheless, these autoantigens might induce a strong autoimmune response leading to severe destruction within the CNS. To investigate the influence of a dominantly presented autoantigen on experimental autoimmune encephalomyelitis (EAE), we generated transgenic mice expressing the autoantigenic peptide MBP 1-10 covalently bound to the MHC class II molecule I-Au. These mice were crossed either with B10.PL or with TCR-transgenic Tg4 mice, specific for the transgenic peptide-MHC combination. In double transgenic mice we found strong thymic deletion and residual peripheral T cells were refractory to antigen stimulation in vitro. Residual peripheral CD4+ T cells expressed activation markers and a high proportion was CD25 positive. Transfer of both CD25-negative and CD25 positive CD4+ T cells from double transgenic animals into B10.PL mice strongly inhibited the progression of EAE. Despite this thorough tolerance induction, some double transgenic mice developed severe signs of EAE after an extended period of time. Our data show that in the circumstances where autoantigenic priming persists, and where the number of antigen-specific T cells is high enough, autoimmunity may prevail over very potent tolerance-inducing mechanisms. PMID- 16361317 TI - A humanized anti-human Fas antibody, R-125224, induces apoptosis in type I activated lymphocytes but not in type II cells. AB - Fas-mediated apoptosis plays an important role in the immune system, including the elimination of autoreactive lymphoid cells. The Fas-mediated signaling pathway is classified into two types, type I and type II, in human lymphoid cell lines. We investigated whether a humanized anti-human Fas mAb, R-125224, has cell selectivity in induction of apoptosis. R-125224 induced apoptosis in H9 cells, SKW6.4 cells and activated human lymphocytes when cross-linked with anti-human IgG. On the other hand, R-125224 did not induce apoptosis in HPB-ALL cells, Jurkat cells or human hepatocytes. By analysis of death-inducing signaling complex formation, it was demonstrated that R-125224 induced apoptosis selectively in type I cells but not in type II cells. Type I cells also expressed more Fas and had more Fas-clustering activity than type II cells. Moreover, co localization of these clusters and GM1, which is an sphingoglycolipid associated with lipid rafts, was detected. It was also shown that R-125224 treatment could reduce the number of activated human CD3+Fas+ cells in a SCID mouse model in vivo. Thus, we demonstrated that R-125224 induces apoptosis specifically in type I cells in vitro and in vivo. PMID- 16361318 TI - Identification of core functional region of murine IL-4 using peptide phage display and molecular modeling. AB - Murine IL-4 is a pleiotropic cytokine with undefined core functional region for eliciting downstream signaling. We used molecular modeling to predict the binding sites recognized by an anti-IL-4-neutralizing mAb (11B.11) and peptide phage display to delineate their makeup. The results of these approaches were confirmed by site-directed mutagenesis analysis. The results suggest that the amino acid residues spanning from 79 to 86 (QRLFRAFR) on IL-4 are of the major binding site for 11B.11. Furthermore, the functional experiments demonstrate that the residues R80, R83 and R86, which are located in the helix C of murine IL-4, play a crucial role in binding to the IL-4R alpha-chain. Taken together, a new core functional region of murine IL-4 is identified, which provides new insight into the interaction between IL-4 and IL-4Ralpha. In addition, the results demonstrate that 11B.11 binds to a core functional region of murine IL-4, which prevents this cytokine from interacting with its cognate receptor. PMID- 16361319 TI - Activation of Vgamma9Vdelta2 T cells by non-peptidic antigens induces the inhibition of subgenomic HCV replication. AB - Hepatitis C virus (HCV) has evolved complex strategies to evade host immune responses and establish chronic infection. Since human Vgamma9Vdelta2 T lymphocytes play a critical role in the immune response against viruses, we analyzed their antiviral functions on Huh7 hepatoma cells carrying the subgenomic HCV replicon (Rep60 cells). In a transwell culture system, Rep60 cells were co cultured with either PBMCs or highly purified gammadelta T cells stimulated by non-peptidic antigens. Vgamma9Vdelta2 T cell activation was associated with a dramatic reduction of HCV RNA levels. Neutralizing antibodies targeting IFN-gamma revealed a critical role for this cytokine in the inhibition of HCV replication. Interestingly, drugs already in clinical use, such as Phosphostim and Zoledronate, known to activate gammadelta T cells, were shown to induce the inhibition of HCV replication mediated by Vgamma9Vdelta2 T cells of HCV patients. Our data suggest that the therapeutic activation of Vgamma9Vdelta2 T lymphocytes may represent an additional strategy to inhibit HCV replication and to restore a Th1-oriented immune response in HCV-infected patients. PMID- 16361320 TI - Cytosolic dehydroascorbate reductase is important for ozone tolerance in Arabidopsis thaliana. AB - Dehydroascorbate reductase (DHAR) is a key component of the ascorbate recycling system. Three functional DHAR genes are encoded in the Arabidopsis genome. Ozone exposure increased the expression of the cytosolic DHAR (cytDHAR) gene alone. We characterized an Arabidopsis mutant with a deficient cytDHAR. The mutant completely lacked cytDHAR activity and was highly ozone sensitive. The amounts of total ascorbate and glutathione were similar in both lines, but the amount of apoplastic ascorbate in the mutant was 61.5% lower. These results indicate that the apoplastic ascorbate, which is generated through the reduction of DHA by cytDHAR, is important for ozone tolerance. PMID- 16361321 TI - Accumulation of the two transcripts of the N gene, conferring resistance to tobacco mosaic virus, is probably important for N gene-dependent hypersensitive cell death. AB - The N gene is a Toll/interleukin-1 receptor (TIR)-nucleotide-binding site (NBS) leucine-rich repeat (LRR)-type resistance (R) gene that generates two alternative transcripts, N(S) and N(L). N(S) encodes the full-length N protein while N(L) is predicted to encode a truncated form of the protein lacking most of the LRR region. We found that the two transcripts were accumulated at 20 degrees C, a permissive temperature, but not at 30 degrees C, a non-permissive temperature for the N gene, in tobacco mosaic virus (TMV)-inoculated leaves. When N gene dependent cell death was triggered by transient 20 degrees C treatment for 2-6 h, considerable levels of the transcripts were accumulated just before cell death, although the levels of N(S) were always higher. The accumulation was induced by transient expression of the 50 kDa helicase domain (p50) of TMV replicase which is the Avr component of N, but not by transient expression of NtMEK2 (DD) mediated cell death or N gene-independent hypersensitive cell death. These results suggest that the accumulation of N(S) and N(L) is associated with the function of N and, above a certain threshold, triggers N-mediated hypersensitive cell death. PMID- 16361322 TI - Dipyridamole echocardiography test in patients with normal or near normal coronary arteries. PMID- 16361323 TI - New evidence of anti-inflammatory interventions in atrial fibrillation. PMID- 16361324 TI - Long-term outcome after surgery for pulmonary stenosis (a longitudinal study of 22-33 years). AB - AIMS: Long-term (>20 years) survival and clinical outcome are only partly documented in patients who underwent surgical repair for isolated pulmonary stenosis. Yet, such data are of critical importance for the future perspectives, medical care, employability, and insurability of these patients. METHODS AND RESULTS: Ninety consecutive patients underwent surgery for pulmonary stenosis between 1968 and 1980 at the Thoraxcenter. A systematic follow-up study was performed in 1990 and again in 2001. Survival after 25 years was 93%. Re intervention was necessary in 15% of the patients, mainly for pulmonary regurgitation. Right atrial and ventricular dilatation and paradoxical septal motion were associated with the need for reoperation. No major ventricular arrhythmias occurred. Supraventricular arrhythmias occurred, only in patients with severe pulmonary regurgitation and disappeared after reoperation. At last follow-up, 67% of the patients was in NYHA Class I and maximal exercise capacity was 90% of normal. Moderate or severe pulmonary regurgitation was present in 37% of the patients. CONCLUSION: Although long-term survival and quality of life are good, pulmonary regurgitation is found in a third of the patients 22-33 years after surgical repair for isolated pulmonary stenosis and reoperation for pulmonary regurgitation was necessary in 9%, especially after the transannular patch technique. PMID- 16361325 TI - Myocardial asynchronism is a determinant of changes in functional mitral regurgitation severity during dynamic exercise in patients with chronic heart failure due to severe left ventricular systolic dysfunction. AB - AIMS: Functional mitral regurgitation (MR) and myocardial asynchronism occur commonly in patients with dilated cardiomyopathy and affect adversely their prognosis and symptoms. The aim of this study was to evaluate the mechanisms of changes in MR severity during dynamic exercise in patients with chronic heart failure (CHF). METHODS AND RESULTS: Seventy patients with CHF due to left ventricular (LV) systolic dysfunction [LV ejection fraction (EF) <40%] and functional MR were studied. All were in sinus rhythm. Medications were left unchanged for the study. Each patient performed a maximal symptom-limited exercise test with continuous 2D-Doppler echocardiography. Mitral regurgitant volume (RV) and effective regurgitant orifice (ERO) were determined at rest and during exercise. LV asynchrony using Doppler tissue imaging and interventricular asynchrony using conventional pulsed-Doppler were evaluated at rest. Resting LV EF averaged 25+/-8%. Mean resting LV and interventricular mechanical delays were 56+/-50 and 43+/-37 ms, respectively. The overall median values for mitral ERO and RV did not significantly change during dynamic exercise (11 [7-16] vs. 11 [6 21] mm2 and 14 [10-22] vs. 12 [9-23] mL, respectively). However, changes in mitral ERO and RV were individually variable and significantly correlated with the degree of LV asynchronism (r=0.66, P<0.0001 and r=0.66, P<0.0001, respectively). CONCLUSION: Changes in MR are variable during dynamic exercise. LV asynchronism at rest substantially contributes to worsening of functional MR during dynamic exercise in patients with CHF due to LV systolic dysfunction. PMID- 16361326 TI - Residual coronary dissections after drug-eluting stenting: the good, the bad, and the ugly. PMID- 16361327 TI - The myth of thin patients as explanation for the excellent results of the Japanese technique in the surgical treatment of gastric cancer. PMID- 16361328 TI - Efficacy of nitazoxanide and paromomycin in biliary tract cryptosporidiosis in an immunosuppressed gerbil model. AB - OBJECTIVES: To evaluate the efficacy of nitazoxanide and paromomycin in biliary tract cryptosporidiosis in an immunosuppressed Mongolian gerbil (Meriones unguiculatus) model. METHODS: Gerbils (1-month-old) were dexamethasone immunosuppressed for 10 days and challenged orally with 10(5) Cryptosporidium parvum oocysts. From day 0 to day 12 post-infection, one group (n=14) was treated with 200 mg/kg/day nitazoxanide and another (n=15) with 100 mg/kg/day paromomycin. Infection and efficacy of nitazoxanide and paromomycin were assessed by measuring oocyst shedding in faeces, biliary tract and ileum histological examination. RESULTS: In nitazoxanide-treated and paromomycin-treated groups as compared with untreated animals (P<0.05), oocyst shedding was partially suppressed in a similar manner (P>0.05). Parasites were present in histological sections of the ileal mucosa of 16/16 infected untreated animals versus 3/14 and 6/15 in the nitazoxanide-treated and the paromomycin-treated groups, respectively (P<0.05). In addition, gall bladder infection was less frequent in nitazoxanide treated (2/14, P<0.01) and paromomycin-treated (5/15, P=0.07) animals than in untreated controls (9/16). No histological alteration of biliary mucosa was observed in both treated and untreated infected gerbils. CONCLUSIONS: Present data support the efficacy of nitazoxanide and, to a lesser extent, paromomycin on biliary C. parvum infection in gerbils, and prompt further investigation of the potential clinical benefits of nitazoxanide in treating human biliary cryptosporidiosis. PMID- 16361329 TI - Water-soluble amphotericin B-polyvinylpyrrolidone complexes with maintained antifungal activity against Candida spp. and Aspergillus spp. and reduced haemolytic and cytotoxic effects. AB - OBJECTIVES: Poor solubility and toxicity severely hinder the clinical use of amphotericin B (AmB), in spite of its attractive chemotherapeutic properties. Water-soluble complexes of AmB and polyvinylpyrrolidone (AmB-PVP) could display lower cytotoxicity while maintaining antifungal activity. METHODS: AmB-PVP [with PVP of 10, 24 and 40 kDa (AC1, AC2 and AC4)] were compared with free AmB for (i) activity against Candida spp. (five albicans; nine non-albicans) and Aspergillus spp. (four strains), (ii) haemolysis of sheep red blood cells, and (iii) release of lactate dehydrogenase from J774 macrophages [with further comparison with free PVP and a liposomal formulation of amphotericin (AmBisome)]. RESULTS: MICs and MFCs of AC1, AC2 and AC4 against Candida spp. and of AC2 and AC4 against Aspergillus spp. were similar to those of AmB (and even lower for some Candida strains). Killing kinetics (24 h) were also similar. Haemolytic activity of AC2 and AC4 was 2-fold lower than that of free AmB. Cytotoxicity of AC2 towards J774 macrophages was 8-fold lower, and that of AC4 5-fold lower than that of AmB and not significantly different from that of AmBisome. The lower cytotoxicity of AC2, AC4 was correlated with a lower cellular accumulation of amphotericin. Spectroscopic analysis shows that the lower toxicity of AmB-PVP was not owing to significant change in the monomeric/polymeric forms ratio of the drug. CONCLUSIONS: AmB-PVP complexes compared favourably with AmB for antifungal activity, were less haemolytic and cytotoxic than AmB, and show a similar cytotoxicity profile to AmBisome. PMID- 16361330 TI - Daptomycin treatment of Staphylococcus aureus experimental chronic osteomyelitis. AB - BACKGROUND: Infection due to methicillin-resistant Staphylococcus aureus (MRSA) is increasingly common in nosocomial and community settings. Daptomycin is a cyclic lipopeptide anti-infective with activity against MRSA, approved for treatment of complicated skin and skin structure infections. Daptomycin may be useful in systemic or local treatment of chronic osteomyelitis. METHODS: We measured mechanical strength of daptomycin- and vancomycin-loaded polymethylmethacrylate (PMMA), assayed in vivo release of daptomycin and vancomycin from daptomycin- and vancomycin-loaded PMMA, respectively, and compared the efficacy of two systemic doses of daptomycin with that of vancomycin, each with or without the respective anti-infective loaded into PMMA, using a rat model of MRSA chronic osteomyelitis. RESULTS: Neither tensile nor compressive strength of PMMA was impacted by impregnation with these antimicrobials at a concentration of 7.5% by weight. The peak concentrations of daptomycin and vancomycin in rat tibial bone surrounding a 7.5% daptomycin- and vancomycin-loaded 3 mm PMMA bead were 178 and 49 mg/L, respectively. In the treatment of experimental osteomyelitis, rats assigned to no treatment, daptomycin 50 mg/kg subcutaneously twice daily, daptomycin 60 mg/kg subcutaneously twice daily, and vancomycin 50 mg/kg intraperitoneally twice daily had 6.4, 4.1, 4.0 and 4.5 median log10 cfu/g of bone at the end of 21 days of therapy. All systemic anti-infectives studied were more active than was no treatment. Daptomycin- or vancomycin-loaded PMMA did not, however, exhibit microbiological efficacy alone or adjunctively, as assessed 21 days after implantation. CONCLUSIONS: Daptomycin is released from PMMA in vivo at a rate similar to that of vancomycin. Systemic daptomycin is as active as vancomycin in a rat model of chronic MRSA experimental osteomyelitis. PMID- 16361331 TI - Single-molecule unfolding force distributions reveal a funnel-shaped energy landscape. AB - The protein folding process is described as diffusion on a high-dimensional energy landscape. Experimental data showing details of the underlying energy surface are essential to understanding folding. So far in single-molecule mechanical unfolding experiments a simplified model assuming a force-independent transition state has been used to extract such information. Here we show that this so-called Bell model, although fitting well to force velocity data, fails to reproduce full unfolding force distributions. We show that by applying Kramers' diffusion model, we were able to reconstruct a detailed funnel-like curvature of the underlying energy landscape and establish full agreement with the data. We demonstrate that obtaining spatially resolved details of the unfolding energy landscape from mechanical single-molecule protein unfolding experiments requires models that go beyond the Bell model. PMID- 16361332 TI - Diffusional encounter of barnase and barstar. AB - We present an analysis of trajectories from Brownian dynamics simulations of diffusional protein-protein encounter for the well-studied system of barnase and barstar. This analysis reveals details about the optimal association pathways, the regions of the encounter complex, possible differences of the pathways for dissociation and association, the coupling of translational and rotation motion, and the effect of mutations on the trajectories. We found that a small free energy barrier divides the energetically most favorable region into a region of the encounter complex above the barnase binding interface and a region around a second energy minimum near the RNA binding loop. When entering the region of the encounter complex from the region near the RNA binding loop, barstar has to change its orientation to increase the electrostatic attraction between the proteins. By concentrating the analysis on the successful binding trajectories, we found that the region of the second minimum is not essential for the binding of barstar to barnase. Nevertheless, this region may be helpful to steer barstar into the region of the encounter complex. When applying the same analysis to several barnase mutants, we found that single mutations may drastically change the free-energy landscape and may significantly alter the population of the two minima. Therefore, certain protein-protein pairs may require careful adaptation of the positions of encounter and transition states when interpreting mutation effects on kinetic rates of association and/or dissociation. PMID- 16361333 TI - Brownian dynamics simulation of knot diffusion along a stretched DNA molecule. AB - Manipulation of individual DNA molecules by optical tweezers has made it possible to tie these molecules into knots. After stretching the DNA molecules the knots become highly localized. In their recent study, Quake and co-authors investigated diffusion of such knots along stretched DNA molecules. We used these data to test the accuracy of a Brownian dynamics simulation of DNA bending motion. We simulated stretched DNA molecules with knots 3(1), 4(1), and 7(1), and determined their diffusion coefficients. Comparison of the simulated and experimental results shows that Brownian dynamics simulation is capable of predicting the rates of large-scale DNA rearrangements within a factor of 2. PMID- 16361334 TI - A sticky chain model of the elongation and unfolding of Escherichia coli P pili under stress. AB - A model of the elongation of P pili expressed by uropathogenic Escherichia coli exposed to stress is presented. The model is based upon the sticky chain concept, which is based upon Hooke's law for elongation of the layer-to-layer and head-to tail bonds between neighboring units in the PapA rod and a kinetic description of the opening and closing of bonds, described by rate equations and an energy landscape model. It provides an accurate description of the elongation behavior of P pili under stress and supports a hypothesis that the PapA rod shows all three basic stereotypes of elongation/unfolding: elongation of bonds in parallel, the zipper mode of unfolding, and elongation and unfolding of bonds in series. The two first elongation regions are dominated by a cooperative bond opening, in which each bond is influenced by its neighbor, whereas the third region can be described by individual bond opening, in which the bonds open and close randomly. A methodology for a swift extraction of model parameters from force-versus elongation measurements performed under equilibrium conditions is derived. Entities such as the free energy, the stiffness, the elastic elongation, the opening length of the various bonds, and the number of PapA units in the rod are determined. PMID- 16361336 TI - Can conformational change be described by only a few normal modes? AB - We suggest a simple method to assess how many normal modes are needed to map a conformational change. By projecting the conformational change onto a subspace of the normal-mode vectors and using root mean square deviation as a test of accuracy, we find that the first 20 modes only contribute 50% or less of the total conformational change in four test cases (myosin, calmodulin, NtrC, and hemoglobin). In some allosteric systems, like the molecular switch NtrC, the conformational change is localized to a limited number of residues. We find that many more modes are necessary to accurately map this collective displacement. In addition, the normal-mode "spectra" can provide useful information about the details of the conformational change, especially when comparing structures with different bound ligands, in this case, calmodulin. Indeed, this approach presents normal-mode analysis as a useful basis in which to capture the mechanism of conformational change, and shows that the number of normal modes needed to capture the essential collective motions of atoms should be chosen according to the required accuracy. PMID- 16361335 TI - Statistical-mechanical theory of DNA looping. AB - The lack of a rigorous analytical theory for DNA looping has caused many DNA-loop mediated phenomena to be interpreted using theories describing the related process of DNA cyclization. However, distinctions in the mechanics of DNA looping versus cyclization can have profound quantitative effects on the thermodynamics of loop closure. We have extended a statistical mechanical theory recently developed for DNA cyclization to model DNA looping, taking into account protein flexibility. Notwithstanding the underlying theoretical similarity, we find that the topological constraint of loop closure leads to the coexistence of multiple classes of loops mediated by the same protein structure. These loop topologies are characterized by dramatic differences in twist and writhe; because of the strong coupling of twist and writhe within a loop, DNA looping can exhibit a complex overall helical dependence in terms of amplitude, phase, and deviations from uniform helical periodicity. Moreover, the DNA-length dependence of optimal looping efficiency depends on protein elasticity, protein geometry, and the presence of intrinsic DNA bends. We derive a rigorous theory of loop formation that connects global mechanical and geometric properties of both DNA and protein and demonstrates the importance of protein flexibility in loop-mediated protein DNA interactions. PMID- 16361337 TI - Structure, surface interactions, and compressibility of bacterial S-layers through scanning force microscopy and the surface force apparatus. AB - Two-dimensional crystalline bacterial surface layers (S-layers) are found in a broad range of bacteria and archaea as the outermost cell envelope component. The self-assembling properties of the S-layers permit them to recrystallize on solid substrates. Beyond their biological interest as S-layers, they are currently used in nanotechnology to build supramolecular structures. Here, the structure of S layers and the interactions between them are studied through surface force techniques. Scanning force microscopy has been used to study the structure of recrystallized S-layers from Bacillus sphaericus on mica at different 1:1 electrolyte concentrations. They give evidence of the two-dimensional organization of the proteins and reveal small corrugations of the S-layers formed on mica. The lattice parameters of the S-layers were a=b=14 nm, gamma=90 degrees and did not depend on the electrolyte concentration. The interaction forces between recrystallized S-layers on mica were studied with the surface force apparatus as a function of electrolyte concentration. Force measurements show that electrostatic and steric interactions are dominant at long distances. When the S-layers are compressed they exhibit elastic behavior. No adhesion between recrystallized layers takes place. We report for the first time, to our knowledge, the value of the compressibility modulus of the S-layer (0.6 MPa). The compressibility modulus is independent on the electrolyte concentration, although loads of 20 mN m-1 damage the layer locally. Control experiments with denatured S proteins show similar elastic properties under compression but they exhibit adhesion forces between proteins, which were not observed in recrystallized S layers. PMID- 16361338 TI - Fluctuation analysis of Caulobacter crescentus adhesion. AB - The aquatic bacterium Caulobacter crescentus divides asymmetrically to a flagellated swarmer cell and a cell with a stalk. At the end of the stalk is an adhesive organelle known as the holdfast, which the stalked cell uses to attach to a solid surface. Often there are two or more cells with their stalks attached to the same holdfast. By analyzing the fluctuations in the stalk angle for a pair of cells attached to a single holdfast, we determine the elastic stiffness of the holdfast. We model the holdfast as three torsional springs in series and find that the effective torsional spring constant for the holdfast is of the order of (10(-17)-10(-18)) Nm, with unequal spring constants. The asymmetry suggests the sequence in which the cells attach to each other, and in some cases suggests that strong crosslinks form between the stalks as they make a shared holdfast. PMID- 16361339 TI - Dehydration of ribonucleotides catalyzed by ribonucleotide reductase: the role of the enzyme. AB - This article focuses on the second step of the catalytic mechanism for the reduction of ribonucleotides catalyzed by the enzyme Ribonucleotide Reductase (RNR). This step corresponds to the protonation/elimination of the substrate's C 2' hydroxyl group. Protonation is accomplished by the neighbor Cys-225, leading to the formation of one water molecule. This is a very relevant step since most of the known inhibitors of this enzyme, which are already used in the fight against certain forms of cancer, are 2'-substituted substrate analogs. Even though some theoretical studies have been performed in the past, they have modeled the enzyme with minimal gas-phase models, basically represented by a part of the side chain of the relevant amino acids, disconnected from the protein backbone. This procedure resulted in a limited accuracy in the position and/or orientation of the participating residues, which can result in erroneous energetics and even mistakes in the choice of the correct mechanism for this step. To overcome these limitations we have used a very large model, including a whole R1 model with 733 residues plus the substrate and 10 A thick shell of water molecules, instead of the minimal gas-phase models used in previous works. The ONIOM method was employed to deal with such a large system. This model can efficiently account for the restrained mobility of the reactive residues, as well as the long-range enzyme-substrate interactions. The results gave additional information about this step, which previous small models could not provide, allowing a much clearer evaluation of the role of the enzyme. The interaction energy between the enzyme and the substrate along the reaction coordinate and the substrate steric strain energy have been obtained. The conclusion was that the barrier obtained with the present model was very similar to the one previously determined with minimal gas-phase models. Therefore, the role of the enzyme in this step was concluded to be mainly entropic, rather than energetic, by placing the substrate and the two reactive residues in a position that allows for the highly favorable concerted trimolecular reaction, and to protect the enzyme radical from the solvent. PMID- 16361340 TI - Importance of the CMAP correction to the CHARMM22 protein force field: dynamics of hen lysozyme. AB - The recently developed CMAP correction to the CHARMM22 force field (C22) is evaluated from 25 ns molecular dynamics simulations on hen lysozyme. Substantial deviations from experimental backbone root mean-square fluctuations and N-H NMR order parameters obtained in the C22 trajectories (especially in the loops) are eliminated by the CMAP correction. Thus, the C22/CMAP force field yields improved dynamical and structural properties of proteins in molecular dynamics simulations. PMID- 16361341 TI - Low-threshold exocytosis induced by cAMP-recruited CaV3.2 (alpha1H) channels in rat chromaffin cells. AB - We have studied the functional role of CaV3 channels in triggering fast exocytosis in rat chromaffin cells (RCCs). CaV3 T-type channels were selectively recruited by chronic exposures to cAMP (3 days) via an exchange protein directly activated by cAMP (Epac)-mediated pathway. Here we show that cAMP-treated cells had increased secretory responses, which could be evoked even at very low depolarizations (-50, -40 mV). Potentiation of exocytosis in cAMP-treated cells did not occur in the presence of 50 microM Ni2+, which selectively blocks T-type currents in RCCs. This suggests that the "low-threshold exocytosis" induced by cAMP is due to increased Ca2+ influx through cAMP-recruited T-type channels, rather than to an enhanced secretion downstream of Ca2+ entry, as previously reported for short-term cAMP treatments (20 min). Newly recruited T-type channels increase the fast secretory response at low voltages without altering the size of the immediately releasable pool. They also preserve the Ca2+ dependence of exocytosis, the initial speed of vesicle depletion, and the mean quantal size of single secretory events. All this indicates that cAMP-recruited CaV3 channels enhance the secretory activity of RCCs at low voltages by coupling to the secretory apparatus with a Ca2+ efficacy similar to that of already existing high threshold Ca2+ channels. Finally, using RT-PCRs we found that the fast inactivating low-threshold Ca2+ current component recruited by cAMP is selectively associated to the alpha1H (CaV3.2) channel isoform. PMID- 16361342 TI - Fatty acid binding proteins: same structure but different binding mechanisms? Molecular dynamics simulations of intestinal fatty acid binding protein. AB - Fatty acid binding proteins (FABPs) carry fatty acids (FAs) and other lipids in the cellular environment, and are thus involved in processes such as FA uptake, transport, and oxidation. These proteins bind either one or two ligands in a binding site, which appears to be inaccessible from the bulk. Thus, the entry of the substrate necessitates a conformational change, whose nature is still unknown. A possible description of the ligand binding process is given by the portal hypothesis, which suggests that the FA enters the protein through a dynamic area known as the portal region. On the other hand, recent simulations of the adipocyte lipid binding protein (ALBP) suggested a different entry site (the alternative portal). In this article, we discuss molecular dynamics simulations of the apo-intestinal-FABP (I-FABP) in the presence of palmitate molecule(s) in the simulation box. The simulations were carried out to study whether the FA can enter the protein during the simulations (as in the ALBP) and where the ligand entry site is (the portal region, the alternative portal or a different domain). The analysis of the simulations revealed a clear difference between the ALBP and the I-FABP. In the latter case, the palmitate preferentially adsorbed to the portal region, which was more mobile than the rest of the protein. However, no ligand entry was observed in the multi-nanosecond-long simulations, in contrast to ALBP. These findings suggest that, although the main structural motif of the FABPs is common, the fine details of each individual protein structure grossly modulate its reactivity. PMID- 16361343 TI - SNARE-mediated lipid mixing depends on the physical state of the vesicles. AB - Reconstitution experiments have suggested that N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) proteins constitute a minimal membrane fusion machinery but have yielded contradictory results, and it is unclear whether the mechanism of membrane merger is related to the stalk mechanism that underlies physiological membrane fusion. Here we show that reconstitution of solubilized neuronal SNAREs into preformed 100 nm liposomes (direct method) yields proteoliposomes with more homogeneous sizes and protein densities than the standard reconstitution method involving detergent cosolubilization of proteins and lipids. Standard reconstitutions yield slow but efficient lipid mixing at high protein densities and variable amounts of lipid mixing at moderate protein densities. However, the larger, more homogenous proteoliposomes prepared by the direct method yield almost no lipid mixing at moderate protein densities. These results suggest that the lipid mixing observed for standard reconstitutions is dominated by the physical state of the membrane, perhaps due to populations of small vesicles (or micelles) with high protein densities and curvature stress created upon reconstitution. Accordingly, changing membrane spontaneous curvature by adding lysophospholipids inhibits the lipid mixing observed for standard reconstitutions. Our data indicate that the lipid mixing caused by high SNARE densities and/or curvature stress occurs by a stalk mechanism resembling the mechanism of fusion between biological membranes, but the neuronal SNAREs are largely unable to induce lipid mixing at physiological protein densities and limited curvature stress. PMID- 16361344 TI - Confinement effects on the thermodynamics of protein folding: Monte Carlo simulations. AB - The effects of chaperonin-like cage-induced confinement on protein stability have been studied for molecules of varying sizes and topologies. Minimalist models based on Go-like interactions are employed for the proteins, and density-of states-based Monte Carlo simulations are performed to accurately characterize the thermodynamic transitions. This method permits efficient sampling of conformational space and yields precise estimates of free energy and entropic changes associated with protein folding. We find that confinement-driven stabilization is not only dependent on protein size and cage radius, but also on the specific topology. The choice of the confining potential is also shown to have an effect on the observed stabilization and the scaling behavior of the stabilization with respect to the cage size. PMID- 16361345 TI - Coarse-grained modeling of the actin filament derived from atomistic-scale simulations. AB - A coarse-grained (CG) procedure that incorporates the information obtained from all-atom molecular dynamics (MD) simulations is presented and applied to actin filaments (F-actin). This procedure matches the averaged values and fluctuations of the effective internal coordinates that are used to define a CG model to the values extracted from atomistic MD simulations. The fluctuations of effective internal coordinates in a CG model are computed via normal-mode analysis (NMA), and the computed fluctuations are matched with the atomistic MD results in a self consistent manner. Each actin monomer (G-actin) is coarse-grained into four sites, and each site corresponds to one of the subdomains of G-actin. The potential energy of a CG G-actin contains three bonds, two angles, and one dihedral angle; effective harmonic bonds are used to describe the intermonomer interactions in a CG F-actin. The persistence length of a CG F-actin was found to be sensitive to the cut-off distance of assigning intermonomer bonds. Effective harmonic bonds for a monomer with its third nearest neighboring monomers are found to be necessary to reproduce the values of persistence length obtained from all-atom MD simulations. Compared to the elastic network model, incorporating the information of internal coordinate fluctuations enhances the accuracy and robustness for a CG model to describe the shapes of low-frequency vibrational modes. Combining the fluctuation-matching CG procedure and NMA, the achievable time- and length scales of modeling actin filaments can be greatly enhanced. In particular, a method is described to compute the force-extension curve using the CG model developed in this work and NMA. It was found that F-actin is easily buckled under compressive deformation, and a writhing mode is developed as a result. In addition to the bending and twisting modes, this novel writhing mode of F-actin could also play important roles in the interactions of F-actin with actin-binding proteins and in the force-generation process via polymerization. PMID- 16361346 TI - Bistability, stochasticity, and oscillations in the mitogen-activated protein kinase cascade. AB - Signaling pathways respond to stimuli in a variety of ways, depending on the magnitude of the input and the physiological status of the cell. For instance, yeast can respond to pheromone stimulation in either a binary or graded fashion. Here we present single cell transcription data indicating that a transient binary response in which all cells eventually become activated is typical. Stochastic modeling of the biochemical steps that regulate activation of the mitogen activated protein kinase Fus3 reveals that this portion of the pathway can account for the graded-to-binary conversion. To test the validity of the model, genetic approaches are used to alter expression levels of Msg5 and Ste7, two of the proteins that negatively and positively regulate Fus3, respectively. Single cell measurements of the genetically altered cells are shown to be consistent with predictions of the model. Finally, computational modeling is used to investigate the effects of protein turnover on the response of the pathway. We demonstrate that the inclusion of protein turnover can lead to sustained oscillations of protein concentrations in the absence of feedback regulation. Thus, protein turnover can profoundly influence the output of a signaling pathway. PMID- 16361347 TI - Correlated fluorescence-atomic force microscopy of membrane domains: structure of fluorescence probes determines lipid localization. AB - Coupling atomic force microscopy (AFM) with high-resolution fluorescence microscopy is an attractive means of identifying membrane domains by both physical topography and fluorescence. We have used this approach to study the ability of a suite of fluorescent molecules to probe domain structures in supported planar bilayers. These included BODIPY-labeled ganglioside, sphingomyelin, and three new cholesterol derivatives, as well as NBD-labeled phosphatidylcholine, sphingomyelin, and cholesterol. Interestingly, many fluorescent lipid probes, including derivatives of known raft-associated lipids, preferentially partitioned into topographical features consistent with nonraft domains. This suggests that the covalent attachment of a small fluorophore to a lipid molecule can abolish its ability to associate with rafts. In addition, the localization of one of the BODIPY-cholesterol derivatives was dependent on the lipid composition of the bilayer. These data suggest that conclusions about the identification of membrane domains in supported planar bilayers on the basis of fluorescent lipid probes alone must be interpreted with caution. The combination of AFM with fluorescence microscopy represents a more rigorous means of identifying lipid domains in supported bilayers. PMID- 16361348 TI - Subcellular in vivo 1H MR spectroscopy of Xenopus laevis oocytes. AB - In vivo magnetic resonance (MR) spectra are typically obtained from voxels whose spatial dimensions far exceed those of the cells they contain. This study was designed to evaluate the potential of localized MR spectroscopy to investigate subcellular phenomena. Using a high magnetic field and a home-built microscopy probe with large gradient field strengths, we achieved voxel sizes of (180 microm)3. In the large oocytes of the frog Xenopus laevis, this was small enough to allow the recording of the first compartment-selective in vivo MR spectra from the animal and vegetal cytoplasm as well as the nucleus. The two cytoplasmic regions differed in their lipid contents and NMR lineshape characteristics differences that are not detectable with whole-cell NMR techniques. In the nucleus, the signal appeared to be dominated by water, whereas other contributions were negligible. We also used localized spectroscopy to monitor the uptake of diminazene acturate, an antitrypanosomal agent, into compartments of a single living oocyte. The resulting spectra from the nucleus and cytoplasm revealed different uptake kinetics for the two components of the drug and demonstrate that MR technology is on the verge of becoming a tool for cell biology. PMID- 16361349 TI - SARS coronavirus E protein in phospholipid bilayers: an x-ray study. AB - We investigated the structure of the hydrophobic domain of the severe acute respiratory syndrome E protein in model lipid membranes by x-ray reflectivity and x-ray scattering. In particular, we used x-ray reflectivity to study the location of an iodine-labeled residue within the lipid bilayer. The label imposes spatial constraints on the protein topology. Experimental data taken as a function of protein/lipid ratio P/L and different swelling states support the hairpin conformation of severe acute respiratory syndrome E protein reported previously. Changes in the bilayer thickness and acyl-chain ordering are presented as a function of P/L, and discussed in view of different structural models. PMID- 16361350 TI - Geographic aspects of poverty and health in Tanzania: does living in a poor area matter? AB - Previous studies have consistently found an inverse relationship between household-level poverty and health status. However, what is not well understood is whether and how the average economic status at the community level plays a role in the poverty-health relationship. The purpose of this study is to investigate the concentration of poverty at the community level in Tanzania and its association with the availability and quality of primary health care services, the utilization of services, and health outcomes among household categories defined by wealth scores. A principal component method has been applied to rank households separately by urban/rural location using reported levels of asset ownership and living conditions. The household wealth scores were also used to classify communities into three cluster-types based on the proportion of households belonging to the poorest wealth tercile. On average, all the wealth terciles living in low poverty concentration areas were found to have better health outcomes and service utilization rates than their counterparts living in high poverty concentration clusters. Consistent with the finding is that high poverty concentration areas were further away from facilities offering primary health care than low poverty concentration areas. Moreover, the facilities closest to the high poverty concentration areas had fewer doctors, medical equipment and drugs. Among the high poverty concentration clusters, the 10 communities with the best women's body mass index (BMI) measures were found to have access to facilities with a greater availability of equipment and drugs than the 10 communities with the worst BMI measures. Although this study does not directly measure quality, the characteristics that differentiate high poverty concentration clusters from low poverty concentration clusters point to quality as more important than physical access among the study population. PMID- 16361351 TI - Salutary effects of estrogen receptor-beta agonist on lung injury after trauma hemorrhage. AB - Although 17beta-estradiol (E2) administration after trauma-hemorrhage attenuates lung injury in male rodents, it is not known whether the salutary effects are mediated via estrogen receptor (ER)-alpha or ER-beta. We hypothesized that the salutary effects of E2 lung are mediated via ER-beta. Male Sprague-Dawley rats underwent trauma-hemorrhage (mean blood pressure 40 mmHg for 90 min, then resuscitation). E2 (50 microg/kg), ER-alpha agonist propyl pyrazole triol (PPT; 5 microg/kg), ER-beta agonist diarylpropiolnitrile (DPN; 5 microg/kg), or vehicle (10% DMSO) was injected subcutaneously during resuscitation. At 24 h after trauma hemorrhage or sham operation, bronchoalveolar fluid (BALF) was collected for protein concentration, LDH activity, and nitrate/nitrite and IL-6 levels. Moreover, lung tissue was used for inducible nitric oxide synthase (iNOS) mRNA/protein expression, nitrate/nitrite and IL-6 levels, and wet/dry weight ratio (n = 6 rats/group). One-way ANOVA and Tukey's test were used for statistical analysis. The results indicated that E2 downregulated lung iNOS expression after trauma-hemorrhage. Protein concentration, LDH activity, and nitrate/nitrite and IL-6 levels in BALF and nitrate/nitrite and IL-6 levels in the lung increased significantly after trauma-hemorrhage; however, administration of DPN but not PPT significantly improved all parameters. Moreover, DPN treatment attenuated trauma-hemorrhage-mediated increase in iNOS mRNA/protein expression in the lung. In contrast, no significant change in the above parameters was observed with PPT. Thus the salutary effects of E2 on attenuation of lung injury are mediated via ER-beta, and ER-beta-induced downregulation of iNOS likely plays a significant role in the DPN-mediated lung protection after trauma-hemorrhage. PMID- 16361352 TI - Genetic and environmental influences of surfactant protein D serum levels. AB - The collectin surfactant protein D (SP-D) is an important component of the pulmonary innate immune system, but SP-D is also present on extrapulmonary epithelial surfaces and in serum, where it has been used as a biomarker for pulmonary disease states. In this study, we investigate the mechanisms defining the constitutional serum level of SP-D and determine the magnitude of the genetic contribution to serum SP-D in the adult population. Recent studies have demonstrated that serum SP-D concentrations in children are genetically determined and that a single nucleotide polymorphism (SNP) located in the NH(2) terminal region (Met11Thr) of the mature protein is significantly associated with the serum SP-D levels. A classic twin study was performed on a twin population including 1,476 self-reported healthy adults. The serum SP-D levels increased with male sex, age, and smoking status. The intraclass correlation was significantly higher for monozygotic (MZ) twin pairs than for dizygotic (DZ) twin pairs. Serum SP-D variance was influenced by nonshared environmental effects and additive genetic effects. Multivariate analysis of MZ and DZ covariance matrixes showed significant genetic correlation among serum SP-D and metabolic variables. The Met11Thr variant explained a significant part of the heritability indicating that serum SP-D variance could be decomposed into non-shared environmental effects (e(2) = 0.19), additive genetic effects (h(2) = 0.42), and the effect of the Met11Thr variations (q(2) = 0.39). PMID- 16361353 TI - Effects of HSP70.1/3 gene knockout on acute respiratory distress syndrome and the inflammatory response following sepsis. AB - Heat shock response has been implicated in attenuating NF-kappaB activation and inflammation following sepsis. Studies utilizing sublethal heat stress or chemical enhancers to induce in vivo HSP70 expression have demonstrated survival benefit after experimental sepsis. However, it is likely these methods of manipulating HSP70 expression have effects on other stress proteins. The aim of this study was to evaluate the role of specific deletion of HSP70.1/3 gene expression on ARDS, NF-kappaB activation, inflammatory cytokine expression, and survival following sepsis. To address this question, we induced sepsis in HSP70.1/3 KO and HSP70.1/3 WT mice via cecal ligation and puncture (CLP). We evaluated lung tissue NF-kappaB activation and TNF-alpha protein expression at 1 and 2 h, IL-6 protein expression at 1, 2, and 6, and lung histopathology 24 h after sepsis initiation. Survival was assessed for 5 days post-CLP. NF-kappaB activation in lung tissue was increased in HSP70.1/3((-/-)) mice at all time points after sepsis initiation. Deletion of HSP70.1/3 prolonged NF-kappaB binding/activation in lung tissue. Peak expression of lung TNF-alpha at 1 and 2 h was also significantly increased in HSP70.1/3((-/-)) mice. Expression of IL-6 was significantly increased at 2 and 6 h, and histopathology revealed a significant increase in lung injury in HSP70.1/3((-/-)) mice. Last, deletion of the HSP70 gene led to increased mortality 5 days after sepsis initiation. These data reveal that absence of HSP70 alone can significantly increase ARDS, activation of NF kappaB, and inflammatory cytokine response. The specific absence of HSP70 gene expression also leads to increased mortality after septic insult. PMID- 16361354 TI - TLR7 ligand prevents allergen-induced airway hyperresponsiveness and eosinophilia in allergic asthma by a MYD88-dependent and MK2-independent pathway. AB - Asthma is one of the leading causes of childhood hospitalization, and its incidence is on the rise throughout the world. Currently, the standard treatment for asthma is the use of corticosteroids to try to suppress the inflammatory reaction taking place in the bronchial tree. Using a murine model of atopic allergic asthma employing a methacholine-hyperresponsive (A/J) as well as a hyporesponsive (C57BL/6) strain of mice sensitized and challenged with ovalbumin, we show that treatment with a synthetic Toll-like receptor 7 (TLR7) ligand (S 28463, a member of the imidazoquinoline family) prevents development of the asthmatic phenotype. Treatment with S-28463 resulted in a reduction of airway resistance and elastance following ovalbumin sensitization and challenge. This was accompanied by a dramatic reduction in infiltration of leukocytes, especially eosinophils, into the lungs of both C57BL/6 and A/J mice following OVA challenge. Treatment with S-28463 also abolished both the elevation in serum IgE level as well as the induction of IL-4, IL-5, and IL-13 by OVA challenge. The protective effects of S-28463 were also observed in MK2 knockout, but not MYD88 knockout, mice. We did not observe a switch in cytokine profile from T(H)2 to T(H)1, as both IL-12p70 and IFN-gamma levels were reduced following S-28463 treatment. These results clearly demonstrate the anti-inflammatory effect of imidazoquinolines in an allergic asthma model as well as the clinical potential of TLR7 ligands in the treatment of allergic diseases. PMID- 16361355 TI - Estrogen receptor regulation of pulmonary alveolar dimensions: alveolar sexual dimorphism in mice. AB - Female rats and mice have smaller and, per body mass (BM), more alveoli and alveolar surface area (Sa) than males of their respective species. This sexual dimorphism becomes apparent about the time of sexual maturity. It is prevented in rats (not tested in mice) by ovariectomy at age 3 wk. In female mice, estrogen receptor (ER)-alpha and ER-beta are required for formation of alveoli of appropriate size and number. We now report the average volume of an alveolus (va) and the number of alveoli per body mass (Na/BM) were not statistically different between ER-alpha(-/-) and wild type (wt) males. However, the combination of a larger value for va and a smaller value for Na/BM, though neither parameter achieved a statistically significant intergroup difference, resulted in a statistically significant lower Sa/BM in ER-alpha(-/-) males compared with wt males. In ER-beta(-/-) males, va was bigger and Na/BM and Sa/BM were lower compared with wt males. Wt males had larger alveoli and lower Na/BM and Sa/BM than wt females. The wt sexual dimorphism of va, Na/BM, and Sa/BM was absent in ER-alpha(-/-) mice. Alveolar size did not differ between ER-beta(-/-) females and males but Na/BM and Sa/BM were greater in ER-beta(-/-) females than in ER-beta(-/ ) males. The results in male mice, with prior findings in female mice, 1) demonstrate estrogen receptors have a smaller effect on alveolar dimensions in male than female mice, 2) show ER-alpha and ER-beta are required for the sexual dimorphism of alveolar size, and 3) show ER-alpha is needed for the sexual dimorphism of body mass-specific alveolar number and surface area. PMID- 16361356 TI - Adenosine A2A receptors promote adenosine-stimulated wound healing in bronchial epithelial cells. AB - Adenosine produces a wide variety of physiological effects through the activation of specific adenosine receptors (A(1), A(2A), A(2B), A(3)). Adenosine, acting particularly at the A(2A) adenosine receptor (A(2A)AR), is a potent endogenous anti-inflammatory agent and sensor of inflammatory tissue damage. The complete healing of wounds is the final step in a highly regulated response to injury. Recent studies on epidermal wounds have identified the A(2A)AR as the main adenosine receptor responsible for altering the kinetics of wound closure. We hypothesized that A(2A)AR promotes wound healing in bronchial epithelial cells (BECs). To test this hypothesis, the human BEC line BEAS-2B and bovine BECs (BBECs) were used. Real-time RT-PCR of RNA from unstimulated BEAS-2B cells revealed transcriptional expression of A(1), A(2A), A(2B) and A(3) receptors. Western blot analysis of lysates from BEAS-2B cells and BBECs detected a single band at 44.7 kDa in both the BECs, indicating the presence of A(2A)AR. In a wound healing model, we found that adenosine stimulated wound repair in cultured BBECs in a concentration-dependent manner, with an optimal closure rate observed between 4 and 6 h. Similarly, the A(2A)AR agonist 5'-(N cyclopropyl)carboxamidoadenosine (CPCA) augmented wound closure, with a maximal closure rate occurring between 4 and 6 h. Inhibition of A(2A)AR with ZM-241385, a known A(2A)AR antagonist, impeded wound healing. In addition, ZM-241385 also attenuated adenosine-mediated wound repair. Kinase studies revealed that adenosine-stimulated airway repair activates PKA by ligating A(2A)AR. Collectively, the data suggest that the A(2A)AR is involved in BEC adenosine stimulated wound healing and may prove useful in understanding purinergic mediated actions on airway epithelial repair. PMID- 16361357 TI - Downregulation of type II bone morphogenetic protein receptor in hypoxic pulmonary hypertension. AB - Heterozygous mutations in the type II receptor for bone morphogenetic protein (BMPR-II) and dysfunction of BMPR-II have been implicated in patients with primary pulmonary hypertension (PH). To clarify the possible involvement of BMP and BMPR-II in the development of hypoxic PH, the expression of BMP-2, BMPR-II, and their downstream signals were investigated in rat lung under normal and hypoxic conditions by RT-PCR, immunoblot, and immunohistochemical methods. In rats under normal conditions, BMP-2 is localized in the endothelium of the pulmonary artery, whereas BMPR-II is abundantly expressed in the endothelium, smooth muscle cells, and adventitial fibroblasts. After 0.5 and 3 days of exposure to hypoxia, upregulation of BMP-2 was observed in the intrapulmonary arteries. The change was accompanied by activation of its downstream signaling, p38 MAPK, and Erk1/2 MAPK, and the apoptotic process, measured by caspase-3 activity and TdT-mediated dUTP nick end labeling-positive cells. In contrast, a significant decrease in the expression of BMPR-II and inactivation of p38 MAPK and caspase-3 were observed in the pulmonary vasculature after 7-21 days of hypoxia exposure. Because BMP-2 is known to inhibit proliferation of vascular smooth muscle cells and promote cellular apoptosis, disruption of BMP signaling pathway through downregulation of BMPR-II in chronic hypoxia may result in pulmonary vascular remodeling due to the failure of critical antiproliferative/differentiation programs in the pulmonary vasculature. These results suggest abrogation of BMP signaling may be a common molecular pathogenesis in the development of PH with various pathophysiological events, including primary and hypoxic PH. PMID- 16361358 TI - Differential protease, innate immunity, and NF-kappaB induction profiles during lung inflammation induced by subchronic cigarette smoke exposure in mice. AB - Cigarette smoke exposure is a major determinant of adverse lung health, but the molecular processes underlying its effects on inflammation and immunity remain poorly understood. Therefore, we sought to understand whether inflammatory and host defense determinants are affected during subchronic cigarette smoke exposure. Dose-response and time course studies of lungs from Balb/c mice exposed to smoke generated from 3, 6, and 9 cigarettes/day for 4 days showed macrophage- and S100A8-positive neutrophil-rich inflammation in lung tissue and bronchoalveolar lavage (BAL) fluid, matrix metalloproteinase (MMP) and serine protease induction, sustained NF-kappaB translocation and binding, and mucus cell induction but very small numbers of CD3+CD4+ and CD3+CD8+ lymphocytes. Cigarette smoke had no effect on phospho-Akt but caused a small upregulation of phospho Erk1/2. Activator protein-1 and phospho-p38 MAPK could not be detected. Quantitative real-time PCR showed upregulation of chemokines (macrophage inflammatory protein-2, monocyte chemoattractant protein-1), inflammatory mediators (TNF-alpha, IL-1beta), leukocyte growth and survival factors [granulocyte-macrophage colony-stimulating factor, colony-stimulating factor (CSF)-1, CSF-1 receptor], transforming growth factor-beta, matrix-degrading MMP-9 and MMP-12, and Toll-like receptor (TLR)2, broadly mirroring NF-kappaB activation. No upregulation was observed for MMP-2, urokinase-type plasminogen activator, tissue-type plasminogen activator, and TLRs 3, 4, and 9. In mouse strain comparisons the rank order of susceptibility was Balb/c > C3H/HeJ > 129SvJ > C57BL6. Partition of responses into BAL macrophages vs. lavaged lung strongly implicated macrophages in the inflammatory responses. Strikingly, except for IL 10 and MMP-12, macrophage and lung gene profiles in Balb/c and C57BL/6 mice were very similar. The response pattern we observed suggests that subchronic cigarette smoke exposure may be useful to understand pathogenic mechanisms triggered by cigarette smoke in the lungs including inflammation and alteration of host defense. PMID- 16361359 TI - Clathrin-mediated endocytosis of FITC-albumin in alveolar type II epithelial cell line RLE-6TN. AB - We examined mechanisms of FITC-albumin uptake by alveolar type II epithelial cells using cultured RLE-6TN cells. Alkaline phosphatase activity and the expression of cytokeratin 19 mRNA, which are characteristic features of alveolar type II epithelial cells, were detected in RLE-6TN cells. The uptake of FITC albumin by the cells was time and temperature dependent and showed the saturation kinetics of high- and low-affinity transport systems. FITC-albumin uptake was inhibited by native albumin, by chemically modified albumin, and by metabolic inhibitors and bafilomycin A(1), an inhibitor of vacuolar H(+)-ATPase. Confocal laser scanning microscopic analysis after FITC-albumin uptake showed punctate localization of fluorescence in the cells, which was partly localized in lysosomes. FITC-albumin taken up by the cells gradually degraded over time, as shown by fluoroimage analyzer after SDS-PAGE. The uptake of FITC-albumin by RLE 6TN cells was not inhibited by nystatin, indomethacin, or methyl-beta cyclodextrin (inhibitors of caveolae-mediated endocytosis) but was inhibited by phenylarsine oxide and chlorpromazine (inhibitors of clathrin-mediated endocytosis) in a concentration-dependent manner. Uptake was also inhibited by potassium depletion and hypertonicity, conditions known to inhibit clathrin mediated endocytosis. These results indicate that the uptake of FITC-albumin in cultured alveolar type II epithelial cells, RLE-6TN, is mediated by clathrin mediated but not by caveolae-mediated endocytosis, and intracellular FITC-albumin is gradually degraded in lysosomes. Possible receptors involved in this endocytic system are discussed. PMID- 16361360 TI - Cigarette smoke disrupts VEGF165-VEGFR-2 receptor signaling complex in rat lungs and patients with COPD: morphological impact of VEGFR-2 inhibition. AB - VEGF is fundamental in the development and maintenance of the vasculature. VEGF(165) signaling through VEGF receptor (VEGFR)-2/kinase insert domain receptor (KDR) is a highly regulated process involving the formation of a tertiary complex with glypican (GYP)-1 and neuropilin (NRP)-1. Both VEGF and VEGFR-2 expression are reduced in emphysematous lungs; however, the mechanism of regulation of VEGF(165) signaling through the VEGFR-2 complex in response to cigarette smoke exposure in vivo, and in smokers with and without chronic obstructive pulmonary disease (COPD), is still unknown. We hypothesized that cigarette smoke exposure disrupts the VEGF(165)-VEGFR-2 complex, a potential mechanism in the pathogenesis of emphysema. We show that cigarette smoke exposure reduces NRP-1 and GYP-1 as well as VEGF and VEGFR-2 levels in rat lungs and that VEGF, VEGFR-2, GYP-1, and NRP-1 expression in the lungs of both smokers and patients with COPD are also reduced compared with nonsmokers. Moreover, our data suggest that specific inhibition of VEGFR-2 alone with NVP-AAD777 would appear not to result in emphysema in the adult rat lung. As both VEGF(165) and VEGFR-2 expression are reduced in emphysematous lungs, decreased GYP-1 and NRP-1 expression may yet further disrupt VEGF(165)-VEGFR-2 signaling. Whether or not this by itself is critical for inducing endothelial cell apoptosis and decreased vascularization of the lung seen in emphysema patients is still unclear at present. However, targeted therapies to restore VEGF(165)-VEGFR-2 complex may promote endothelial cell survival and help to ameliorate emphysema. PMID- 16361361 TI - ATP stimulates MMP-2 release from human aortic smooth muscle cells via JNK signaling pathway. AB - Aortic smooth muscle cell release of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) has been implicated in aortic aneurysm pathogenesis, but proximal modulation of release is poorly understood. Extracellular nucleotides regulate vascular smooth muscle cell metabolism in response to physiochemical stresses, but nucleotide modulation of MMP and/or TIMP release has not been reported. We hypothesized that nucleotides modulate MMP-2 and TIMP-2 release from human aortic smooth muscle cells (HASMCs) via distinct purinergic receptors and signaling pathways. We exposed HASMCs to exogenous ATP and other nucleotides with and without interleukin-1beta (IL-1beta). HASMCs were pretreated in some experiments with apyrase, which degrades ATP, and inhibitors of ERK1/2, JNK, and p38 MAPK. MMP-2 and TIMP-2 released into supernatant were assessed using ELISA and Western blotting. ATP, adenosine, and UTP significantly stimulated MMP-2 release in the presence of IL-1beta (300 nM ATP: 181 +/- 22%, P = 0.003; 30 microm adenosine: 244 +/- 150%, P = 0.001; and 200 microm UTP: 153 +/ 40%, P = 0.015; vs. 100% constitutive). ATP also stimulated MMP-2 release in the absence of IL-1beta (100 microm ATP: 148 +/- 38% vs. 100% constitutive). Apyrase significantly reduced ATP-stimulated MMP-2 release (apyrase + 500 nM ATP: 59 +/- 3% vs. 124 +/- 7% with 500 nM ATP). Rank-order agonist potency for MMP-2 release was consistent with ATP activation of PAY and PAY receptors. ATP induced phosphorylation of intracellular JNK, and inhibition of the JNK pathway blocked ATP-stimulated MMP-2 release, indicating signaling via this pathway. Nucleotides are thus novel stimulants of MMP-2 release from HASMCs and may provide a mechanistic link between physiochemical stress in the aorta and aneurysms, especially in the context of inflammation. PMID- 16361362 TI - Flow regulates intercellular communication in HAEC by assembling functional Cx40 and Cx37 gap junctional channels. AB - Direct cell-to-cell transfer of ions and small signaling molecules via gap junctions plays a key role in vessel wall homeostasis. Vascular endothelial gap junctional channels are formed by the connexin (Cx) proteins Cx37, Cx40, and Cx43. The mechanisms regulating connexin expression and assembly into functional channels have not been fully identified. We investigated the dynamic regulation of endothelial gap junctional intercellular communication (GJIC) by fluid flow and the participation of each vascular connexin in functional human endothelial gap junctions in vitro. Human aortic endothelial cells (HAEC) were exposed for 5, 16, and 24 h to physiological flows in a parallel-plate flow chamber. Connexin protein expression and localization were evaluated by immunocytochemistry, and functional GJIC was evaluated by dye injection. Connexin-mimetic peptide inhibitors were used to assess the specific connexin composition of functional channels. HAEC monolayers in culture exhibited baseline functional communication at a striking low level despite abundant expression of Cx43 and Cx40 localized at cell-to-cell appositions. Upon exposure to flow, GJIC by dye spread demonstrated a significant time-dependent increase from baseline levels, reaching 7.5-fold in 24 h. Inhibition studies revealed that this response was mediated primarily by Cx40, with lesser contributions of the other two vascular connexins assembled into functional homotypic and/or heterotypic channels. This is the first study to demonstrate that flow simultaneously and differentially regulates expression of the Cx37, Cx40, and Cx43 proteins and their involvement in the augmentation of intercellular communication by dye transfer in human endothelial cells in vitro. PMID- 16361363 TI - Ability of short-time Fourier transform method to detect transient changes in vagal effects on hearts: a pharmacological blocking study. AB - Conventional spectral analyses of heart rate variability (HRV) have been limited to stationary signals and have not allowed the obtainment of information during transient autonomic cardiac responses. In the present study, we evaluated the ability of the short-time Fourier transform (STFT) method to detect transient changes in vagal effects on the heart. We derived high-frequency power (HFP, 0.20 0.40 Hz) as a function of time during active orthostatic task (AOT) from the sitting to standing posture before and after selective vagal (atropine sulfate 0.04 mg/kg) and sympathetic (metoprolol 0.20 mg/kg) blockades. The HFP minimum point during the first 30 s after standing up was calculated and compared with sitting and standing values. Reactivity scores describing the fast and slow HFP responses to AOT were calculated by subtracting the minimum and standing values from the sitting value, respectively. The present results, obtained without controlled respiration, showed that in the drug-free condition, HFP decreased immediately after standing up (P < 0.001) and then gradually increased toward the level characteristic for the standing posture (P < 0.001), remaining lower than in the sitting baseline posture (P < 0.001). The magnitudes of the fast and slow HFP responses to AOT were abolished by the vagal blockade (P < 0.001) and unaffected by the sympathetic blockade. These findings indicate that HFP derived by the STFT method provided a tool for monitoring the magnitude and time course of transient changes in vagal effects on the heart without the need to interfere with normal control by using blocking drugs. PMID- 16361364 TI - Testosterone enhances early cardiac remodeling after myocardial infarction, causing rupture and degrading cardiac function. AB - Cardiac rupture can be fatal after myocardial infarction (MI). Experiments in animals revealed gender differences in rupture rate; however, patient data are controversial. We found a significantly higher rupture rate in testosterone treated female mice within 1 wk after MI, whereas castration in males significantly reduced rupture. We hypothesized that testosterone may adversely affect remodeling after MI, exaggerating the inflammatory response and increasing cardiac rupture, whereas estrogen may be cardioprotective, attenuating early remodeling and reducing rupture rate. We studied the effect of gender and hormone manipulation on morphological and histological changes during early remodeling after MI in 4-wk-old male and female C57BL/6J mice and how these events could affect cardiac function. Females were randomly divided into 1) sham ovariectomy + placebo (s-ovx + P), 2) s-ovx + testosterone (T), 3) ovx + P, and 4) ovx + T; males were divided into 1) sham castration + P (s-cas + P), 2) s-cas + 17beta estradiol (E), 3) cas + P, and 4) cas + E. At 6 wk after gonadectomy and hormone manipulation, MI was induced. Mice were randomly killed 1, 2, 4, 7, and 14 days after MI. The left ventricle was weighed and sectioned for evaluation of MI size, infarct expansion index (IEI), and neutrophil infiltration. Transthoracic echocardiography was performed in conscious mice in the 14-day group before organ harvest. Cardiac rupture rate and IEI were significantly higher in testosterone treated females and noncastrated males than in controls; these effects were accompanied by enhanced neutrophil infiltration and pronounced deterioration of cardiac function and left ventricular dilatation. Ovariectomy in females and estrogen supplementation in males did not confer significant protection from cardiac rupture, IEI, or neutrophil infiltration. We concluded that, in mice, high testosterone levels enhance acute myocardial inflammation, adversely affecting myocardial healing and early remodeling, as indicated by increased cardiac rupture, and possibly causing deterioration of cardiac function after MI, and, conversely, estrogen seems to have no significant protective effect in the acute phase after MI. PMID- 16361365 TI - Probing lipid rafts with proximity imaging: actions of proatherogenic stimuli. AB - Glycosylphosphatidylinositol (GPI)-anchored proteins have been shown to cluster in microdomains enriched in glycosphingolipids and cholesterol and represent a relatively selective marker of lipid rafts. In recent years, several attempts have been made to use fluorescent probes to nondisruptively label these domains in living cells. Here, we have transfected endothelial cells with a GPI-anchored thermotolerant green fluorescent protein (ttGFP) to show colocalization of this fluoroprobe with another marker of lipid rafts, urokinase-type plasminogen activator receptor-1. ttGFP was used to quantify the cell surface area occupied by lipid rafts and to examine the effect of various proatherogenic signals on lipid rafts. Exposure of endothelial cells to asymmetric dimethylarginine and oxidized LDL (oxLDL), as well as oxidant stress, reduced the cell surface area occupied by lipid rafts. Next, the property of ttGFP to undergo a shift in absorbance depending on the clustering of these molecules was utilized to perform proximity imaging (PRIM). PRIM showed that nitric oxide (NO) increased the distance between GPI-anchored ttGFP molecules clustered in lipid-rich microdomains. This "unclustering" of GPI-anchored ttGFP was not reproduced by prooxidant signals and was due to reduction in membrane-cytoskeletal constraints on the lipid rafts. These findings suggested that two fundamentally different mechanisms modulate lipid rafts: 1) substance regulation of lipid rafts involving modification of cholesterol and sphingolipids and 2) structural regulation of lipid rafts through disruption of membrane-cytoskeletal interactions, switching off the spatial confinement of lipid rafts. PMID- 16361366 TI - Effects of modulation of left ventricular contractile state and loading conditions on tissue Doppler myocardial performance index. AB - The Tei index is clinically useful to quantify left ventricular (LV) function, but it requires sequential Doppler recordings from two different views. A related myocardial performance index (MPI) using tissue Doppler (TD) can be rapidly calculated from a single beat; however, its ability to quantify contractility and the effects of acute changes in loading have not been determined. Our aim was to test the hypothesis that TD MPI can quantify contractile state but is affected by acute alterations in loading, using LV pressure-volume relations in an animal model. Eight dogs were studied by using mitral annular TD, high-fidelity pressure, and conductance catheters. TD MPI was calculated as (a' - b')/b', where a' was the duration of mitral annular velocity during diastole and b' was the duration of the systolic wave. End-systolic elastance (Ees), the time constant of isovolumic relaxation (tau), and peak positive and negative first derivative of pressure (dP/dtmax and dP/dtmin, respectively) were used as measures of LV function. Data were obtained at baseline, at dobutamine and esmolol infusion to alter contractile state, and at inferior vena cava and aortic occlusion to alter preload and afterload. TD MPI decreased from 0.83 (SD 0.19) to 0.62 (SD 0.20) with dobutamine and increased to 1.19 (SD 0.26) with esmolol. TD MPI significantly correlated with dP/dtmax (r = -0.76), Ees (r = -0.68), dP/dtmin (r = 0.82), and tau (r = 0.78); however, it was affected by acute decreases in preload [from 0.83 (SD 0.19) to 1.09 (SD 0.36)] and acute increases in afterload [to 1.23 (SD 0.17)]. All the above increases and decreases and r values were significant (P < 0.05 vs. baseline). In conclusion, TD MPI can rapidly quantify alterations in LV contractile state but is affected by acute alterations in preload and afterload. PMID- 16361367 TI - Characterization of human cardiac mitochondrial ATP-sensitive potassium channel and its regulation by phorbol ester in vitro. AB - Activation of the mitochondrial ATP-sensitive K+ channel (mitoKATP) and its regulation by PKC are critical events in preconditioning induced by ischemia or pharmaceutical agents in animals and humans. The properties of the human cardiac mitoKATP channel are unknown. Furthermore, there is no evidence that cytosolic PKC can directly regulate the mitoKATP channel located in the inner mitochondrial membrane (IMM) due to the physical barrier of the outer mitochondrial membrane. In the present study, we characterized the human cardiac mitoKATP channel and its potential regulation by PKC associated with the IMM. IMM fractions isolated from human left ventricles were fused into lipid bilayers in symmetrical potassium glutamate (150 mM). The conductance of native mitoKATP channels was usually below 80 pS ( approximately 70%), which was reduced by ATP and 5-hydroxydecanoic acid (5-HD) in a dose- and time-dependent manner. The native mitoKATP channel is activated by diazoxide and inhibited by ATP and 5-HD. The PKC activator phorbol 12-myristate 13-acetate (2 microM) increased the cumulative open probability of the mitoKATP channel previously inhibited by ATP (P < 0.05), but its inactive analog 4alpha-phorbol 12,13-didecanoate had no effect. Western blot analysis detected an inward rectifying K+ channel (Kir6.2) immunoreactive protein at 56 kDa and PKC-delta in the IMM. These data provide the first characterization of the human cardiac mitoKATP channel and its regulation by PKC(s) in IMM. This local PKC control mechanism may represent an alternative pathway to that proposed previously for cytosolic PKC during ischemic/pharmacological preconditioning. PMID- 16361368 TI - Acute responses to phytoestrogens in small arteries from men with coronary heart disease. AB - The aim of this study was to investigate acute vasodilator responses to phytoestrogens and selective estrogen receptor-alpha (ERalpha) agonist in isolated small arteries from men with established coronary heart disease (CHD) and with a history of myocardial infarction versus healthy male control subjects. As to methodology, small arteries obtained from subcutaneous fat biopsies and mounted on a wire myograph were preconstricted with norepinephrine, and dilator responses to increasing nanomolar-micromolar concentrations of the phytoestrogens resveratrol and genistein (predominantly ERbeta agonists) and to propyl-[1H] pyrazole-1,3,5-triyl-trisplenol (PPT, a selective ERalpha agonist) were determined. These were compared with responses to reference compound 17beta estradiol (17beta-E2). Concentration-response curves were constructed before and after nitric oxide (NO) synthase inhibition with Nomega-nitro-L-arginine methyl ester. As a result, relaxation induced by the investigated compounds was similar in men with CHD and control men, but in both groups PPT and genistein-induced relaxation was greater than that of resveratrol and 17beta-E2. NO contributed to both phytoestrogens and PPT-induced relaxation but not to 17beta-E2 responses in arteries from control men. This NO-mediated component of relaxation was absent in arteries from men with established CHD. In conclusion, phytoestrogens, at concentrations achievable by ingestion of phytoestrogen-rich food products, evoke dilatation ex vivo of small peripheral arteries from normal men and those with established CHD. The contribution of NO to dilatory responses by these compounds is pertinent to arteries from control males, whereas other NO-independent dilatory mechanism(s) are involved in arteries from CHD. PMID- 16361369 TI - Involvement of Rho kinase pathway in the mechanism of renal vasoconstriction and cardiac hypertrophy in rats with experimental heart failure. AB - Rho-dependent kinases serve as downstream effectors of several vasoconstrictor systems, the activities of which are upregulated in congestive heart failure (CHF). We evaluated renal and cardiac effects of Y-27632, a highly selective Rho kinase inhibitor, in an experimental model of volume-overload CHF. Effects of acute administration of Y-27632 (0.3 mg/kg) on renal hemodynamic and clearance parameters and effects of chronic treatment (10.0 mg.kg(-1).day(-1) for 7 days via osmotic minipumps) on cardiac hypertrophy and cumulative Na+ excretion were studied in male Wistar rats with aortocaval fistula and control rats. The Y-27632 induced decrease in renal vascular resistance (from 40.4 +/- 4.6 to 26.0 +/- 3.1 resistance units, P < 0.01) in CHF rats was associated with a significant increase in total renal blood flow (+34%) and cortical and medullary blood flow (approx +37 and +27%, respectively). These values were significantly higher than those in control rats and occurred despite a decrease in mean arterial pressure ( 15 mmHg). Despite the marked renal vasodilatory effect, Y-27632 did not alter glomerular filtration rate and renal Na+ excretion. Chronic administration of Y 27632 did not alter daily or cumulative renal Na+ excretion in CHF rats but was associated with a significant decrease in heart-to-body weight ratio, an index of cardiac hypertrophy: 0.32 +/- 0.007, 0.46 +/- 0.017, and 0.37 +/- 0.006% in control, CHF, and CHF + Y-27632 rats, respectively. The findings suggest that Rho kinase-dependent pathways are involved in the mechanisms of renal vasoconstriction and cardiac hypertrophy in rats with volume-overload heart failure. Selective blockade of these signaling pathways may be considered an additional tool to improve renal perfusion and attenuate cardiac hypertrophy in heart failure. PMID- 16361392 TI - Arabidopsis cytokinin receptor mutants reveal functions in shoot growth, leaf senescence, seed size, germination, root development, and cytokinin metabolism. AB - We used loss-of-function mutants to study three Arabidopsis thaliana sensor histidine kinases, AHK2, AHK3, and CRE1/AHK4, known to be cytokinin receptors. Mutant seeds had more rapid germination, reduced requirement for light, and decreased far-red light sensitivity, unraveling cytokinin functions in seed germination control. Triple mutant seeds were more than twice as large as wild type seeds. Genetic analysis indicated a cytokinin-dependent endospermal and/or maternal control of embryo size. Unchanged red light sensitivity of mutant hypocotyl elongation suggests that previously reported modulation of red light signaling by A-type response regulators may not depend on cytokinin. Combined loss of AHK2 and AHK3 led to the most prominent changes during vegetative development. Leaves of ahk2 ahk3 mutants formed fewer cells, had reduced chlorophyll content, and lacked the cytokinin-dependent inhibition of dark induced chlorophyll loss, indicating a prominent role of AHK2 and, particularly, AHK3 in the control of leaf development. ahk2 ahk3 double mutants developed a strongly enhanced root system through faster growth of the primary root and, more importantly, increased branching. This result supports a negative regulatory role for cytokinin in root growth regulation. Increased cytokinin content of receptor mutants indicates a homeostatic control of steady state cytokinin levels through signaling. Together, the analyses reveal partially redundant functions of the cytokinin receptors and prominent roles for the AHK2/AHK3 receptor combination in quantitative control of organ growth in plants, with opposite regulatory functions in roots and shoots. PMID- 16361393 TI - The Arabidopsis vitamin E pathway gene5-1 mutant reveals a critical role for phytol kinase in seed tocopherol biosynthesis. AB - We report the identification and characterization of a low tocopherol Arabidopsis thaliana mutant, vitamin E pathway gene5-1 (vte5-1), with seed tocopherol levels reduced to 20% of the wild type. Map-based identification of the responsible mutation identified a G-->A transition, resulting in the introduction of a stop codon in At5g04490, a previously unannotated gene, which we named VTE5. Complementation of the mutation with the wild-type transgene largely restored the wild-type tocopherol phenotype. A knockout mutation of the Synechocystis sp PCC 6803 VTE5 homolog slr1652 reduced Synechocystis tocopherol levels by 50% or more. Bioinformatic analysis of VTE5 and slr1652 indicated modest similarity to dolichol kinase. Analysis of extracts from Arabidopsis and Synechocystis mutants revealed increased accumulation of free phytol. Heterologous expression of these genes in Escherichia coli supplemented with free phytol and in vitro assays of recombinant protein produced phytylmonophosphate, suggesting that VTE5 and slr1652 encode phytol kinases. The phenotype of the vte5-1 mutant is consistent with the hypothesis that chlorophyll degradation-derived phytol serves as an important intermediate in seed tocopherol synthesis and forces reevaluation of the role of geranylgeranyl diphosphate reductase in tocopherol biosynthesis. PMID- 16361395 TI - Patients' views on interpersonal continuity in primary care: a sense of security based on four core foundations. AB - BACKGROUND: A deep and comprehensive understanding of what patients value about having a personal doctor in primary care is lacking. OBJECTIVES: To acquire a comprehensive understanding of the core values of having a personal doctor in a continuing doctor-patient relationship in primary care among long-term, chronically ill patients. METHOD: In this qualitative study, 14 chronically ill patients at three primary health care centres were strategically selected. The centres were selected to include patients with experiences from both long-term and short-term doctors. The patients were asked about their views on having a personal doctor in a continuing doctor-patient relationship in primary care compared with having different short-term doctors. Sixteen health care professionals were interviewed about what chronically ill patients convey to them about having a personal doctor in contrast to seeing different short-term locum doctors. The in-depth interviews were transcribed verbatim and analysed by qualitative content analysis. RESULTS: The core category, i.e. a universal concept that many patients used to describe the impact of having access to a personal doctor, was a sense of security. This was based on four main categories or core foundations which were: feelings of coherence, confidence in care, a trusting relationship and accessibility. In turn, the four main categories emerged from two to four of subcategories. CONCLUSION: The foundations that underpin the value of personal care from the patients' perspective could be based on categories found in this study. PMID- 16361394 TI - Different domains control the localization and mobility of LIKE HETEROCHROMATIN PROTEIN1 in Arabidopsis nuclei. AB - Plants possess a single gene for the structurally related HETEROCHROMATIN PROTEIN1 (HP1), termed LIKE-HP1 (LHP1). We investigated the subnuclear localization, binding properties, and dynamics of LHP1 proteins in Arabidopsis thaliana cells. Transient expression assays showed that tomato (Solanum lycopersicum) LHP1 fused to green fluorescent protein (GFP; Sl LHP1-GFP) and Arabidopsis LHP1 (At LHP1-GFP) localized to heterochromatic chromocenters and showed punctuated distribution within the nucleus; tomato but not Arabidopsis LHP1 was also localized within the nucleolus. Mutations of aromatic cage residues that recognize methyl K9 of histone H3 abolished their punctuated distribution and localization to chromocenters. Sl LHP1-GFP plants displayed cell type dependent subnuclear localization. The diverse localization pattern of tomato LHP1 did not require the chromo shadow domain (CSD), whereas the chromodomain alone was insufficient for localization to chromocenters; a nucleolar localization signal was identified within the hinge region. Fluorescence recovery after photobleaching showed that Sl LHP1 is a highly mobile protein whose localization and retention are controlled by distinct domains; retention at the nucleolus and chromocenters is conferred by the CSD. Our results imply that LHP1 recruitment to chromatin is mediated, at least in part, through interaction with methyl K9 and that LHP1 controls different nuclear processes via transient binding to its nuclear sites. PMID- 16361396 TI - The mapping of fine and ultrafine particle concentrations in an engine machining and assembly facility. AB - Aerosol mapping was used to assess particle number and mass concentration in an engine machining and assembly facility in the winter and spring. Number and mass concentration maps were constructed from data collected with two mobile sampling carts, each equipped with a condensation particle counter (10 nm < diameter < 1 microm) and an optical particle counter (300 nm < diameter < 20 microm). Number concentrations inside the facility ranged from 15 to 150 times greater than that outside the facility and were highly dependent on season. The greatest number concentration (>1,000,000 particles cm(-3)) occurred in winter in an area where mass concentration was low (<0.10 mg m(-3)). The increased number of particles was attributed to the exhaust of direct-fire, natural-gas burners used to heat the supply air. The greatest mass concentrations were found around metalworking operations that were poorly enclosed. The larger particles that dominated particle mass in this area were accompanied by ultrafine particles, probably generated through evaporation and subsequent condensation of metalworking fluid components. Repeat mapping events demonstrated that these ultrafine particles persist in workplace air over long time periods. PMID- 16361397 TI - Occupational exposures and non-Hodgkin lymphoma: where do we stand? PMID- 16361398 TI - Physical activity, psychological complaints, and occupational health. PMID- 16361399 TI - Lung cancer risk and talc not containing asbestiform fibres: a review of the epidemiological evidence. AB - A literature search was done and all epidemiological cancer studies mentioning talc as a risk factor were selected. The talc exposed populations were divided into three groups: (1) populations in which no other occupational carcinogen was mentioned (only talc millers satisfied this criterion); (2) populations of talc miners exposed to talc, quartz, and/or radon; and (3) other industrial populations in which talc is associated with quartz, nitrosamines, and asbestos depending on the study. No excess lung cancer mortality was found for the populations of talc millers exposed to high levels of talc but without any other potential carcinogen (SMR = 0.92, 42 cases) while the summary of mortality of talc miners exposed to quartz and/or radon was in excess (fixed effect SMR = 1.20, random effect RR = 1.85, 40 cases). Six studies in other industrial settings were identified. All reported increased lung cancer mortality among talc exposed workers but the talc exposure was confounded with other carcinogens and only one study was able to adjust on them. In conclusion, no increased lung cancer mortality was observed among talc millers despite their high exposure experience. In populations in which talc was associated with other potential carcinogens, some lung cancer excesses were observed. PMID- 16361400 TI - Can strenuous leisure time physical activity prevent psychological complaints in a working population? AB - AIMS: To investigate the longitudinal relation between strenuous leisure time physical activity and psychological complaints (depression and emotional exhaustion) in a Dutch working population in order to find evidence for the preventive role of physical activity in the development of psychological complaints. METHODS: All data came from the Study on Musculoskeletal disorders, Absenteeism, Stress, and Health (SMASH), a three year follow up study that started between 1994 and 1995. The study population consisted of 1747 workers from 34 companies. Generalised estimating equation (GEE) analyses were performed to investigate the longitudinal relation between strenuous leisure time physical activity and psychological complaints using models with and without a time lag. Logistic regression analyses were performed to study the relation between physical activity and sickness absence due to psychological complaints during the three year follow up study. RESULTS: Only in workers with a sedentary job was strenuous leisure time physical activity (1-2 times per week) significantly associated with a reduced risk of future depression and emotional exhaustion. This was not the case for physical activity at higher frequencies (> or =3 times per week). There was a dose-response relation between strenuous leisure time physical activity and poor general health which was strongest in workers with a sedentary job. Strenuous leisure time physical activity (1-2 times per week) was associated with a lower risk of long term absenteeism (>21 days), whereas physical activity at a higher frequency was not. CONCLUSIONS: Results suggest that strenuous leisure time physical activity might play a role in the prevention of future psychological complaints, poor general health, and long term absenteeism in a working population. Workers with a sedentary job seem to benefit more from strenuous leisure time physical activity than workers without a sedentary job. PMID- 16361401 TI - Occupation and malignant lymphoma: a population based case control study in Germany. AB - AIMS: To identify occupations suspected to be associated with malignant lymphoma and to generate new hypotheses about occupational risks in a multicentre, population based case control study. METHODS: Male and female patients with malignant lymphoma (n = 710) aged 18-80 years of age were prospectively recruited in six study regions in Germany. For each newly recruited lymphoma case, a sex, region, and age matched control was drawn from the population registers. Odds ratios and 95% confidence intervals for major occupations and industries were calculated using conditional logistic regression analysis, adjusted for smoking (in pack-years) and alcohol consumption. Patients with specific lymphoma subentities were additionally compared with the entire control group using unconditional logistic regression analysis. RESULTS: The following economic/industrial sectors were positively associated with lymphoma: food products, beverages, tobacco; paper products, publishing and printing; and metals. Chemicals; real estate, renting, and business activities were negatively associated with lymphoma diagnosis. The authors observed an increased overall lymphoma risk among architects; maids; farmers; glass formers; and construction workers. Shoemaking and leather goods making was negatively associated with the lymphoma diagnosis (although based on small numbers). In the occupational group analysis of lymphoma subentities, Hodgkin's lymphoma was significantly associated only with rubber and plastic products making; diffuse large B cell lymphoma risk was considerably increased among metal processors; follicular lymphoma showed highly significant risk increases for several occupational groups (medical, dental, and veterinary workers; sales workers; machinery fitters; and electrical fitters); and multiple myeloma showed a particularly pronounced risk increase for farmers as well as for agriculture and animal husbandry workers. CONCLUSIONS: The results partly confirm previously defined occupational risks. Occupational risk factors for follicular lymphomas might differ from the overall risk factors for malignant lymphoma. PMID- 16361402 TI - Trial by fire: a multivariate examination of the relation between job tenure and work injuries. AB - AIMS: This study examined the relation between months on the job and lost-time claim rates, with a particular focus on age related differences. METHODS: Workers' compensation records and labour force survey data were used to compute claim rates per 1000 full time equivalents. To adjust for potential confounding, multivariate analyses included age, sex, occupation, and industry, as well job tenure as predictors of claim rates. RESULTS: At any age, the claim rates decline as time on the job increases. For example, workers in the first month on the job were over four times more likely to have a lost-time claim than workers with over one year in their current job. The job tenure injury associations were stronger among males, the goods industry, manual occupations, and older adult workers. CONCLUSIONS: The present results suggest that all worker subgroups examined show increased risk when new on the job. Recommendations for improving this situation include earlier training, starting workers in low hazard conditions, reducing job turnover rates in firms, and improved monitoring of hazard exposures that new workers encounter. PMID- 16361403 TI - Self perceived health and mental health among women flight attendants. AB - AIMS: The authors investigated associations of work related risk factors with self perceived health as less than "good" and psychological distress among Italian women flight attendants. METHODS: The authors conducted a cross sectional survey on health and mental health among 1955 former and current flight attendants, using a postal questionnaire. RESULTS: More current than former flight attendants reported self perceived health as fair to poor and psychological distress measured as a GHQ-12 score of six or more. Among current flight attendants, reporting health as fair to poor was associated with low job satisfaction (OR 1.89) and recent experiences of sexual harassment by passengers (OR 2.83). Psychological distress was associated with low job satisfaction (OR 2.38) and frequent tension with partner over childcare (OR 1.79). CONCLUSIONS: Perceived health as fair to poor and psychological distress were greater among current flight attendants and were related to job characteristics and family difficulties. Perceived poor health has been shown in the literature to be related to mortality, high job strain, and early retirement, and psychological distress is associated with work absence. The effect of sexual harassment by passengers on perceived health of flight attendants may be relevant to other working women dealing with the public. The health effects of family/work conflicts, low job satisfaction, and sexual harassment should be explored more in depth, using qualitative as well as quantitative methods among working women in various occupations. PMID- 16361404 TI - Occupational risk factors for nasopharyngeal cancer among female textile workers in Shanghai, China. AB - AIMS: To investigate whether occupational exposure to dusts and chemicals in the Chinese textile industry are associated with risk of nasopharyngeal cancer. METHODS: Sixty seven nasopharyngeal carcinoma (NPC) cases identified during 1989 98 and a random sample (n = 3188) of women were included in a case cohort study nested in a cohort of 267,400 women textile workers in Shanghai, China. A complete occupational history of work in the textile industry was obtained for each woman. A job exposure matrix developed by experienced industrial hygienists was used to assess exposures to specific dusts and chemicals. RESULTS: Risk of NPC is associated with cumulative exposure to cotton dust. The hazard ratio for women cumulatively exposed to >143.4 mg/m3 x years of cotton dust was 3.6 (95% CI 1.8 to 7.2) compared with unexposed women. Trends of increasing risk were also found with increasing duration of exposure to acids and caustics (p = 0.05), and with years worked in dyeing processes (p = 0.06). Women who worked at least 10 years in dyeing processes had a 3.6-fold excess risk of NPC (95% CI 1.0 to 12.1). CONCLUSIONS: Occupational exposure to cotton dust, acids, and caustics, and work in dyeing and printing jobs in the textile industry may have increased risk of NPC in this cohort. PMID- 16361405 TI - The role of physical workload and pain related fear in the development of low back pain in young workers: evidence from the BelCoBack Study; results after one year of follow up. AB - AIMS: To study the influence of work related physical and psychosocial factors and individual characteristics on the occurrence of low back pain among young and pain free workers. METHODS: The Belgian Cohort Back Study was designed as a prospective cohort study. The study population of this paper consisted of 716 young healthcare or distribution workers without low back pain lasting seven or more consecutive days during the year before inclusion. The median age was 26 years with an interquartile range between 24 and 29 years. At baseline, these workers filled in a questionnaire with physical exposures, work related psychosocial factors and individual characteristics. One year later, the occurrence of low back pain lasting seven or more consecutive days and some of its characteristics were registered by means of a questionnaire. To assess the respective role of predictors at baseline on the occurrence of low back pain in the following year, Cox regression with a constant risk period for all subjects was applied. RESULTS: After one year of follow up, 12.6% (95% CI 10.1 to 15.0) of the 716 workers had developed low back pain lasting seven or more consecutive days. An increased risk was observed for working with the trunk in a bent and twisted position for more than two hours a day (RR 2.2, 95% CI 1.2 to 4.1), inability to change posture regularly (RR 2.1, 95% CI 1.3 to 3.5), back complaints in the year before inclusion (RR 1.7, 95% CI 1.1 to 2.8), and high scores of pain related fear (RR 1.8, 95% CI 1.0 to 3.1). Work related psychosocial factors and physical factors during leisure time were not predictive. CONCLUSION: This study highlighted the importance of physical work factors and revealed the importance of high scores of pain related fear in the development of low back pain among young workers. PMID- 16361406 TI - Laboratory work and pregnancy outcomes: a study within the National Birth Cohort in Denmark. AB - AIMS: To examine pregnancy outcomes in women doing laboratory work. METHODS: Using data from the Danish National Birth Cohort (1997-2003), the authors conducted a prospective cohort study of 1025 female laboratory technicians and 8037 female teachers (as reference). The laboratory technicians were asked about laboratory work tasks during pregnancy in an interview (at around 16 weeks of gestation). Pregnancy outcomes were obtained by linking the cohort to the national registers. Hazard ratios (HRs) of late fetal loss and diagnosing of congenital malformations were calculated by using Cox regression, and odds ratios (ORs) of preterm birth and small for gestational age were calculated by using logistic regression. RESULTS: Overall, there were no significant differences in pregnancy outcomes between laboratory technicians and teachers. However, we found that laboratory technicians working with radioimmunoassay or radiolabelling had an increased risk of preterm birth (OR = 2.2, 95% CI 0.8 to 6.2 for radioimmunoassay, and OR = 1.9, 95% CI 0.8 to 4.6 for radiolabelling) and "major" malformations (HR = 2.1, 95% CI 1.0 to 4.7 for radioimmunoassay, and HR = 1.8, 95% CI 0.9 to 3.7 for radiolabelling). The ORs of preterm birth doubled for women working with these tasks every day or several times a week. When an exposure matrix was applied, an increased risk of "major" malformations for exposure to organic solvents was seen. CONCLUSIONS: The results did not indicate any high risk of reproductive failures in laboratory technicians in general. Exposure to radioisotopes may carry a high risk of preterm birth and congenital malformations. This finding deserves further investigation. PMID- 16361407 TI - Agricultural seed dust as a potential cause of organic dust toxic syndrome. AB - AIMS: Episodes of serious work related health problems resembling organic dust toxic syndrome (ODTS) in workers of a grass seed quality inspection laboratory prompted the authors to study personal endotoxin exposure levels in this facility and in the agricultural seed processing industry. In addition, microbial and inflammatory characteristics of agricultural seeds were studied. METHODS: The authors assessed inhalable dust and endotoxin levels in 101 samples from 57 workers in grass, cereal, and vegetable seed plants who were handling mainly grass seeds as bulk product, and horticulture seeds in smaller quantities. Additionally, real-time dust exposure was measured using a DataRAM monitor in 12 grass seed workers to obtain more information on exposure patterns during specific tasks. Endotoxin concentrations in seed extracts were determined by LAL assay and seed samples were analysed by scanning electron microscopy. Release of inflammatory cytokines was measured in supernatants of whole blood samples stimulated with lipopolysaccharide (LPS) or agricultural seed extracts in a human whole blood assay (WBA). RESULTS: Endotoxin concentrations in personal samples were high (geometric mean 1800 EU/m3), particularly in the grass seed quality inspection lab where endotoxin levels up to 274 000 EU/m3 were measured. The recommended health based endotoxin exposure limit of 50 EU/m3 was amply exceeded in almost all personal samples. Job tasks dumping and mixing were associated with highest dust and endotoxin exposures, which was confirmed by real-time measurements. Microbial infestation was found in almost all seed samples. WBA results showed that most seed extracts were capable of inducing a pronounced dose dependent cytokine release. CONCLUSIONS: Workers handling grass, cereal, or vegetable seeds are at risk of exposure to high levels of endotoxin containing seed dust. Occupational exposure to inhalable agricultural seed dust can induce inflammatory responses, and is a potential cause of ODTS. PMID- 16361408 TI - World at work: hospital pharmacy clean-rooms. PMID- 16361409 TI - Cancer risk in benzene exposed workers. PMID- 16361410 TI - The two sides of the "endotoxin coin". PMID- 16361411 TI - Regeneration of pancreatic islets after partial pancreatectomy in mice does not involve the reactivation of neurogenin-3. AB - Understanding the factors and mechanisms involved in beta-cell regeneration will guide therapeutic efforts to augment beta-cell mass in patients with diabetes. Neurogenin-3 (Ngn3) is a bHLH transcription factor that responds to Notch signaling and whose expression marks endocrine progenitors. During fetal development, all endocrine cells are derived from Ngn3(+) precursors. Although expression of Ngn3 in the adult pancreas has not been reported, it has been suggested that islet regeneration in adult organisms recapitulates embryonic developmental pathways. Here, we investigated whether beta-cell regeneration in adult mice recapitulates the embryonic pathway involving Ngn3 activation. Despite full recovery of beta-cell mass after 50% partial pancreatectomy (Ppx) in BALB/c mice, no pancreatic Ngn3 immunoreactivity was detected, even when the beta-cell trophic glucagon-like peptide-1 receptor agonist exendin-4 was administered after the procedure. Even when we used the stable expression of enhanced green fluorescent protein (EGFP) in Ngn3(EGFP/+) mice to trace Ngn3 expression after Ppx, no pancreatic Ngn3 expression was detected. Although ectopic expression of Ngn3 can promote an endocrine transcriptional program in adult cells and may thus have therapeutic potential in the development of surrogate beta-cells, our studies indicate that a reactivation of endogenous Ngn3 expression is not required for adult beta-cell regeneration in vivo. PMID- 16361412 TI - Dendritic cells pulsed with hepatitis C virus NS3 protein induce immune responses and protection from infection with recombinant vaccinia virus expressing NS3. AB - Infections with Hepatitis C virus (HCV) pose a serious health problem worldwide. In this study, the hypothesis that adoptive transfer of dendritic cells (DCs) pulsed with HCV NS3 protein and matured with an oligodeoxynucleotide (ODN) containing CpG motifs (CpG) ex vivo would initiate potent HCV-specific protective immune responses in vivo was tested. NS3 protein was efficiently transduced into DCs and treatment of DCs with CpG ODN induced phenotypic maturation and specifically increased the expression of CD40. DCs matured with CpG ODN produced higher interleukin 12 levels and a stronger allogeneic T-cell response compared with untreated DCs. Notably, there were no differences between NS3-pulsed DCs and DCs pulsed with a control protein with respect to phenotype, cytokine production or mixed lymphocyte reaction, indicating that transduction with NS3 protein did not impair DC functions. Compared with the untreated NS3-pulsed DCs, the NS3 pulsed DCs matured with CpG ODN induced stronger cellular immune responses including enhanced cytotoxicity, higher interferon-gamma production and stronger lymphocyte proliferation. Upon challenge with a recombinant vaccinia virus expressing NS3, all mice immunized with NS3-pulsed DCs showed a significant reduction in vaccinia virus titres when compared with mock-immunized mice. However, the NS3-pulsed DCs matured with CpG ODN induced higher levels of protection compared with the untreated NS3-pulsed DCs. These data are the first to show that NS3-pulsed DCs induce specific immune responses and provide protection from viral challenge, and also demonstrate that CpG ODNs, which have a proven safety profile, would be useful in the development of DC vaccines. PMID- 16361413 TI - Isolation and characterization of a chimpanzee alphaherpesvirus. AB - Although both beta- and gammaherpesviruses indigenous to great-ape species have been isolated, to date all alphaherpesviruses isolated from apes have proven to be human viruses [herpes simplex virus types 1 (HSV1) and 2 (HSV2) or varicella zoster virus]. If the alphaherpesviruses have co-evolved with their host species, some if not all ape species should harbour their own alphaherpesviruses. Here, the isolation and characterization of an alphaherpesvirus from a chimpanzee (ChHV) are described. Sequencing of a number of genes throughout the ChHV genome indicates that it is collinear with that of HSV. Phylogenetic analyses place ChHV in a clade with HSV1 and HSV2, the alphaherpesviruses of Old World monkeys comprising a separate clade. Analysis of reactivity patterns of HSV2-immune human sera and ChHV-immune chimpanzee sera by competition ELISA support this relationship. Phylogenetic analyses also place ChHV rather than HSV1 as the closest relative of HSV2. PMID- 16361414 TI - Modified vaccinia virus Ankara multiplies in rat IEC-6 cells and limited production of mature virions occurs in other mammalian cell lines. AB - Recombinant viruses based on modified vaccinia virus Ankara (MVA) are vaccine candidates against infectious diseases and cancers. Presently, multiplication of MVA has been demonstrated in chicken embryo fibroblast and baby hamster kidney (BHK-21) cells only. The multiplication and morphogenesis of a recombinant (MVA HANP) and non-recombinant MVA strain in BHK-21 and 12 other mammalian cell lines have now been compared. Rat IEC-6 cells were fully permissive to MVA infection. The virus yield in IEC-6 cells was similar to that obtained in BHK-21 cells at low as well as high multiplicities of infection. Vero cells were semi-permissive to MVA infection. Mature virions were produced in supposedly non-permissive cell lines. The multiplication and morphogenesis of non-recombinant MVA and MVA-HANP were similar. These results are relevant to the production and biosafety of MVA vectored vaccines. PMID- 16361415 TI - Deletion of gene A41L enhances vaccinia virus immunogenicity and vaccine efficacy. AB - Vaccinia virus (VACV) is the vaccine that was used to eradicate smallpox and is being developed as a recombinant vaccine for other pathogens. Removal of genes encoding immunomodulatory proteins expressed by VACV may enhance virus immunogenicity and improve its potential as a vaccine. Protein A41 is a candidate for removal, having sequence similarity to the VACV chemokine-binding protein, vCKBP, and an association with reduced inflammation during dermal infection. Here, it is shown that, at low doses, VACV strain Western Reserve (WR) lacking A41L (vDeltaA41L) was slightly more virulent than wild-type and revertant controls after intranasal infection of BALB/c mice. The primary immune response to vDeltaA41L was marked by an increase in the percentage of VACV-specific gamma interferon-producing CD8(+) T cells and enhancement of cytotoxic T-cell responses in the spleen. However, this augmentation of cellular response was not seen in lung infiltrates. Splenic CD8(+) T-cell responses were also enhanced when VACV strain modified vaccinia virus Ankara (MVA) lacking A41L was used to immunize mice. Lastly, immunization with VACV MVA lacking A41L provided better protection than control viruses to subsequent challenge with a 300 LD(50) dose of VACV WR. This study provides insight into the immunomodulatory role of A41 and suggests that MVA lacking A41 may represent a more efficacious vaccine. PMID- 16361416 TI - Evolution of Hepatitis B virus in an acute hepatitis B patient co-infected with genotypes B and C. AB - The interactions between different genotypes of Hepatitis B virus (HBV) in co infected patients remain largely unknown, especially in acute infection. Here, the evolution of HBV strains was studied in an acute, self-limited hepatitis B patient co-infected with genotypes Ba (B2) and C. Virological analyses were performed at four time points after admission: T1 (5 days), T2 (11 days), T3 (22 days) and T4 (260 days). A dominant-genotype change from genotype C to Ba was found after anti-HBV e antigen (anti-HBe) seroconversion. Further clonal and phylogenetic analyses of the pre-S and pre-core/core regions of HBV were carried out to clarify the interactions between genotypes Ba and C. All clones propagated from T1 and T2 were of genotype C. In contrast, clones propagated from T3 (after anti-HBe seroconversion) were of genotype Ba, C and/or recombinant within the pre S region. At T4, all clones were of genotype Ba with a 123 bp (from nt 3147 of the pre-S1 region to nt 54 of the pre-S2 region) in-frame pre-S deletion and had lost the start codon of the middle envelope protein and the nucleocapsid-binding site. Phylogenetic analysis showed that genetic distance was greater at T3 after seroconversion to anti-HBe. By using SimPlot, the breakpoint of one pre-S recombinant was located at nt 3069-3100 and the other two at nt 49-87. In conclusion, HBV genotype Ba may overtake genotype C as the predominant strain after anti-HBe seroconversion in acute hepatitis B. Recombination within the pre S region emerged transiently and the pre-S deletion mutant was finally cleared. PMID- 16361417 TI - CCAAT/enhancer-binding protein beta represses human papillomavirus 11 upstream regulatory region expression through a promoter-proximal YY1-binding site. AB - CCAAT/enhancer-binding protein beta (C/EBPbeta) can function as a repressor or as an activator of human papillomavirus (HPV) gene expression, depending on which cell type the experiments are conducted. In this report, it was shown that within primary human foreskin keratinocyte cells (HFK) the activity of C/EBPbeta can be switched from that of a repressor of HPV11 expression to an activator by mutating a single promoter-proximal consensus YY1-binding site within the HPV11 upstream regulatory region (URR). It was shown that in HFK cells, exogenous expression of C/EBPbeta significantly activates the expression of mutant HPV11 URR reporter plasmids that contain deletions which overlap a 127 bp region (-269 to -142). Inclusive in this region are binding sites for multiple transcription factors, including AP1, YY1 and C/EBPalpha. Only mutation of the YY1 site resulted in the switch in phenotype, indicating that C/EBPbeta represses HPV11 expression in these cells via YY1 binding. The level of YY1 activity was also measured in HFK cells transfected with a C/EBPbeta expression plasmid and a significant increase in YY1 activity as compared with mock-transfected cells was found. C33A cells, which exhibit activation of wild-type HPV11 gene expression with exogenous C/EBPbeta co-expression, failed to demonstrate C/EBPbeta-induced YY1 activation. It was concluded that in HFK cells, exogenous C/EBPbeta induces the activity of YY1, which, in turn, can repress HPV11 URR expression through the promoter proximal YY1-binding site. PMID- 16361418 TI - Expression of hepatitis C virus-derived core or NS3 antigens in human dendritic cells leads to induction of pro-inflammatory cytokines and normal T-cell stimulation capabilities. AB - The majority of hepatitis C virus (HCV)-infected individuals become chronically infected, which can result in liver cirrhosis and hepatocellular carcinoma. Patients with chronic HCV are unable to prime and maintain vigorous T-cell responses, which are required to rid the body of the viral infection. Dendritic cells (DCs) are the professional antigen-presenting cells that probably play a dominant role in priming and maintaining vigorous T-cell responses in HCV infection. Furthermore, inefficient DC function may play an important role in HCV chronicity. In order to determine the effect of HCV NS3 and core proteins on phenotype and function of human DCs, recombinant adenoviral vectors containing NS3 or core genes were used to infect human DCs. HCV NS3- or core-protein expression in DCs was confirmed by Western blotting and immunofluorescence staining. The DCs expressing HCV NS3 or core proteins expressed several inflammatory cytokine mRNAs, had a normal phenotype and effectively stimulated allogeneic T cells, as well as T cells specific for another foreign antigen (tetanus toxoid). These findings are important for rational design of cellular vaccine approaches for the immunotherapy of chronic HCV. PMID- 16361419 TI - Population genetic history of hepatitis C virus 1b infection in China. AB - Subtype 1b is the most common strain of Hepatitis C virus (HCV) in China. Here, the molecular epidemiology and epidemic history of this strain were investigated by conducting phylogenetic and population genetic analyses of E1 and NS5B gene sequences sampled from nine Chinese cities. The phylogenetic analysis indicated the presence of two clusters of Chinese strains that did not include reference strains from other countries, suggesting that these clusters represent two independent chains of HCV transmission within China. The remaining Chinese isolates were more closely related to reference strains from other countries. The date of origin and past population dynamics of the two groups were investigated using a new population genetic method, the Bayesian skyline plot. The estimated dates of origin of both groups coincide with the period of the Chinese 'Cultural Revolution' during the years 1966-1976. Both groups grew at a rapid exponential rate between approximately 1970 and approximately 1990, after which transmission slowed considerably. Possible explanations for the groups' fast spread and subsequent slowdown are discussed, including parenteral transmission by unsafe injection, iatrogenic transmission by infected blood or blood products and improvements in blood safety since 1990. These results shed light on HCV transmission in China and may help to predict the future burden of HCV-related disease in the country. PMID- 16361421 TI - A link between translation of the hepatitis C virus polyprotein and polymerase function; possible consequences for hyperphosphorylation of NS5A. AB - Hyperphosphorylation of NS5A is thought to play a key role in controlling hepatitis C virus (HCV) RNA replication. Using a tetracycline-regulable baculovirus delivery system to introduce non-culture-adapted HCV replicons into HepG2 cells, we found that a point mutation in the active site of the viral polymerase, NS5B, led to an increase in NS5A hyperphosphorylation. Although replicon transcripts lacking elements downstream of NS5A also had altered NS5A hyperphosphorylation, this did not explain the changes resulting from polymerase inactivation. Instead, two additional findings may be related to the link between polymerase activity and NS5A hyperphosphorylation. Firstly, we found that disabling polymerase activity, either by targeted mutation of the polymerase active site or by use of a synthetic inhibitor, stimulated translation from the replicon transcript. Secondly, when the rate of translation of non-structural proteins from replicon transcripts was reduced by use of a defective encephalomyocarditis virus internal ribosome entry site, there was a substantial decrease in NS5A hyperphosphorylation, but this was not observed when non structural protein expression was reduced by simply lowering replicon transcript levels using tetracycline. Therefore, one possibility is that the point mutation within the active site of NS5B causes an increase in NS5A hyperphosphorylation because of an increase in translation from each viral transcript. These findings represent the first demonstration that NS5A hyperphosphorylation can be modulated without use of kinase inhibitors or mutations within non-structural proteins and, as such, provide an insight into a possible means by which HCV replication is controlled during a natural infection. PMID- 16361420 TI - Selective transmission of hepatitis C virus genotypes and quasispecies in humans and experimentally infected chimpanzees. AB - This study determined whether selective transmission of hepatitis C virus (HCV) species occurred among human and chimpanzee recipients of contaminated blood products or plasma containing multiple genotypes, subgenotypes and quasispecies. Commercially prepared factor VIII concentrate (lot DO56), produced prior to HCV testing and inactivation, was subsequently found by direct cloning to contain the following subgenotypes: 1a and 1b (73 % of clones), 2a (13 % of clones), 2b (11 % of clones) and 3a (4 % of clones). A patient transfused with factor VIII concentrate DO56 was diagnosed with clinical non-A, non-B hepatitis and subsequently found to be infected with HCV subgenotype 1b. Among five chimpanzees inoculated experimentally with the same factor VIII concentrate, two were infected only with HCV subgenotype 1a and three were infected with approximately equivalent clonal proportions of subgenotypes 1a and 1b. HCV hypervariable region 1 (HVR1) quasispecies analysis of the DO56 factor VIII concentrate and a serum specimen from the single chimpanzee that developed a chronic HCV infection following inoculation with DO56 showed 0-56 % nucleotide variation. However, specimens from chimpanzees infected in the second to fourth passages of the DO56 inoculum had 0-8 % HVR1 quasispecies nucleotide variation. The high HVR1 quasispecies variation in the factor VIII concentrate and its first passage in chimpanzees indicates the presence of multiple HCV isolates, whereas the low variation in the second to fourth chimpanzee passages suggests transmission of a single HCV isolate. These findings strongly suggest selective transmission of HCV isolates during experimental chimpanzee infection and among humans exposed to multiple HCV species. PMID- 16361422 TI - Structural and functional characterization of the coxsackievirus B3 CRE(2C): role of CRE(2C) in negative- and positive-strand RNA synthesis. AB - A stem-loop element located within the 2C-coding region of the coxsackievirus B3 (CVB3) genome has been proposed to function as a cis-acting replication element (CRE). It is shown here that disruption of this structure indeed interfered with viral RNA replication in vivo and abolished uridylylation of VPg in vitro. Site directed mutagenesis demonstrated that the previously proposed enteroviral CRE consensus loop sequence, R(1)NNNAAR(2)NNNNNNR(3), is also applicable to CVB3 CRE(2C) and that a positive correlation exists between the ability of CRE(2C) mutants to serve as template in the uridylylation reaction and the capacity of these mutants to support viral RNA replication. To further investigate the effects of the mutations on negative-strand RNA synthesis, an in vitro translation/replication system containing HeLa S10 cell extracts was used. Similar to the results observed for poliovirus and rhinovirus, it was found that a complete disruption of the CRE(2C) structure interfered with positive-strand RNA synthesis, but not with negative-strand synthesis. All CRE(2C) point mutants affecting the enteroviral CRE consensus loop, however, showed a marked decrease in efficiency to induce negative-strand synthesis. Moreover, a transition (A(5)G) regarding the first templating adenosine residue in the loop was even unable to initiate complementary negative-strand synthesis above detectable levels. Taken together, these results indicate that the CVB3 CRE(2C) is not only required for the initiation of positive-strand RNA synthesis, but also plays an essential role in the efficient initiation of negative-strand RNA synthesis, a conclusion that has not been reached previously by using the cell-free system. PMID- 16361423 TI - Analysis of sequential hepatitis A virus strains reveals coexistence of distinct viral subpopulations. AB - Hepatitis A virus (HAV) is a hepatotropic member of the family Picornaviridae. Despite a remarkable antigenic stability, recent results have shown that HAV exists in vivo and in cell culture as distributions of genetically related, non identical variants, referred to as quasispecies. To gain insight into HAV evolution over time in a specific geographical region, genotype I consensus sequences from strains isolated in France in consecutive years were studied. Phylogenetic neighbour-joining method and a non-hierarchical partition analysis, designed to analyse viral quasispecies, indicate that at least five distinct subpopulations of HAV were identified in the course of the disease episode. Strikingly, over time, different subpopulations cycled in dominance. The coexistence of distinct subpopulations whose frequency varies with time is consistent with quasispecies dynamics, and suggests that variation in the dominant HAV population may provide HAV adaptability without being reflected in significant antigenic variation. PMID- 16361424 TI - Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses. AB - The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 3'-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus isolates to identify species of interest. The four primer pairs were 100 % specific when tested against enterovirus prototype strains and panels of isolates of known serotype (a total of 193 isolates). For evaluation in a typical application, the species-specific primers were used to screen 186 previously uncharacterized non-polio enterovirus isolates. The HEV-B primers amplified 68.3 % of isolates, while the HEV-A and HEV-C primers accounted for 9.7 and 11.3 % of isolates, respectively; no isolates were amplified with the HEV-D primers. Twelve isolates (6.5 %) were amplified by more than one primer set and eight isolates (4.3 %) were not amplified by any of the four primer pairs. Serotypes were identified by partial sequencing of the VP1 capsid gene, and in every case sequencing confirmed that the species-specific PCR result was correct; the isolates that were amplified by more than one species-specific primer pair were mixtures of two (11 isolates) or three (one isolate) species of viruses. The eight isolates that were not amplified by the species-specific primers comprised four new serotypes (EV76, EV89, EV90 and EV91) that appear to be unique members of HEV-A based on VP1, 3D and 3'-non-translated region sequences. PMID- 16361425 TI - Alignment of capsid protein VP1 sequences of all human rhinovirus prototype strains: conserved motifs and functional domains. AB - An alignment was made of the deduced amino acid sequences of the entire capsid protein VP1 of all human rhinovirus (HRV) prototype strains to examine conserved motifs in the primary structure. A set of previously proposed crucially important amino acids in the footprints of the two known receptor molecules was not conserved in a receptor group-specific way. In contrast, VP1 and VP3 amino acids in the minor receptor-group strains corresponding to most of the predicted ICAM-1 footprint definitely differed from those of the ICAM-1-using major receptor-group strains. Previous antiviral-sensitivity classification showed an almost-complete agreement with the species classification and a fair correlation with amino acids aligning in the antiviral pocket. It was concluded that systematic alignment of sequences of related virus strains can be used to test hypotheses derived from molecular studies of individual model viruses and to generate ideas for future studies on virus structure and replication. PMID- 16361426 TI - Broad cellular immunity with robust memory responses to simian immunodeficiency virus following serial vaccination with adenovirus 5- and 35-based vectors. AB - Adenovirus serotype 35 (Ad35) is a promising vaccine platform for human immunodeficiency virus (HIV) infection and emerging infectious diseases as it is uncommon in humans worldwide and is distinct from Ad5, the major vaccine serotype for which many individuals have pre-existing immunity. The immunogenicity of a first-generation, replication-competent Ad35-based vaccine was tested in the simian immunodeficiency virus (SIV) rhesus macaque model by evaluating its capacity to boost immunity generated by Ad5-based vectors. A series of four immunizations with replication-defective Ad5 vectors expressing SIVmac239 gag induced high-frequency responses mediated by both CD8(+) and CD4(+) T cells directed against several epitopes. Ad5-specific neutralizing antibody responses that did not neutralize Ad35 were rapidly induced but waned over time. Subsequent immunization with Ad5-based vectors was minimally effective, whereas immunization with Ad35-based vectors generated a strong increase in the frequency of Gag specific T cells with specificities that were unchanged. While this boosting response was relatively transient, challenge with the distinct pathogenic isolate SIV/DeltaB670 generated robust and selective recall responses to Gag with similar specificities as induced by vaccination that were elevated for 25 weeks relative to controls. Vaccination had measurable albeit minor effects on virus load. Unexpectedly, regional hypervariability within the Gag sequence of SIV/DeltaB670 was associated with mutation of the conserved CD8(+) T-cell epitope CM9 without concurrent flanking mutations and in the absence of immune pressure. These findings support the further development of Ad35 as a vaccine vector, and promote vaccine regimens that utilize serial administration of heterologous adenoviruses. PMID- 16361427 TI - Screening for CD8 cytotoxic T lymphocytes specific for Gag of human immunodeficiency virus type 1 subtype B' Henan isolate from China and identification of novel epitopes restricted by the HLA-A2 and HLA-A11 alleles. AB - The human immunodeficiency virus type 1 (HIV-1) epidemic in China is increasing rapidly at an irrepressible rate. It is caused by HIV-1 subtype B' in central China. After the full-length genome sequencing of the Henan isolate was performed, the definition of optimal cytotoxic T-lymphocyte (CTL) epitopes across the Henan isolate genome has become crucial for vaccine design. In this study, by using ELISPOT assays with synthetic peptides corresponding to the sequence of the Henan isolate, the identification and analysis of Gag-specific CTL responses among 28 treated and 26 untreated infected paid blood donors (PBDs) from the Henan and Hubei provinces of China are presented. These studies focused on CTL responses restricted by the human leukocyte antigen (HLA)-A2 and -A11 molecules, two of the most prominent HLA-A alleles in the Chinese population. The results suggested that, in the subgroup analysis, the magnitude of response in the infected treated subgroup [median, 93 spot-forming cells (SFCs) per 10(6) peripheral blood mononuclear cells (PBMCs)] was significantly lower than that in the chronically infected untreated subgroup (median, 221 SFCs per 10(6) PBMCs), and HLA-A2-restricted treated PBDs had a response of a much higher frequency and magnitude than that of HLA-A11-restricted treated PBDs. Moreover, some novel peptides restricted by the HLA-A2 and -A11 molecules were identified. PMID- 16361428 TI - Structural analysis of the human respiratory syncytial virus phosphoprotein: characterization of an alpha-helical domain involved in oligomerization. AB - Human respiratory syncytial virus (HRSV) phosphoprotein (P), an essential cofactor of the viral polymerase, is much shorter (241 aa) than and has no sequence similarity to P of other paramyxoviruses. Nevertheless, bioinformatic analysis of HRSV P sequence revealed a modular organization, reminiscent of other paramyxovirus Ps, with a central structured domain (aa 100-200), flanked by two intrinsically disordered regions (1-99 and 201-241). To test the predicted structure experimentally, HRSV P was purified from cell extracts infected with recombinant vaccinia virus or HRSV. The estimated molecular mass of P by gel filtration (approximately 500 kDa) greatly exceeded the theoretical mass of a homotetramer, proposed as the oligomeric form of native P. Nevertheless, the profile of cross-linked products obtained with purified P resembled that reported by others with P purified from bacteria or mammalian cells. Thus, the shape of HRSV P probably influences its elution from the gel filtration column, as reported for other paramyxovirus Ps. Digestion of purified HRSV P with different proteases identified a trypsin-resistant fragment (X) that reacted with a previously characterized monoclonal antibody (021/2P). N-terminal sequencing and mass spectrometry analysis placed the X fragment boundaries (Glu-104 and Arg-163) within the predicted structured domain of P. Cross-linking and circular dichroism analyses indicated that fragment X was oligomeric, with a high alpha-helical content, properties resembling those of the multimerization domain of Sendai and rinderpest virus P. These results denote structural features shared by HRSV and other paramyxovirus Ps and should assist in elucidation of the HRSV P structure. PMID- 16361429 TI - Comparison of in vitro replication features of H7N3 influenza viruses from wild ducks and turkeys: potential implications for interspecies transmission. AB - In previous work, it was shown that turkey H7N3 influenza viruses, presumably derived 'in toto' from interspecies transmission of duck viruses in Northern Italy, had only 2 aa differences in haemagglutinin and a few amino acid differences as well as a 23 aa deletion in neuraminidase compared with duck viruses. Here, the replication of these duck and turkey viruses in Madin-Darby canine kidney cells was investigated with respect to virus-cell fusion and viral elution from red blood cells. Duck viruses showed similar receptor-binding properties to turkey viruses but possessed a higher pH of fusion activation than the turkey viruses. Conversely, turkey viruses were not able to elute from red blood cells. These data confirm that neuraminidase-stalk deletion impairs the release of virions from cells and also confirm existence of naturally occurring viruses with different pH fusion activities, raising the possibility that these features may play a role in the evolution of influenza viruses in different hosts. PMID- 16361430 TI - Genetic elements regulating packaging of the Bunyamwera orthobunyavirus genome. AB - The genome of Bunyamwera virus (BUN; family Bunyaviridae, genus Orthobunyavirus) comprises three segments of negative-sense, single-stranded RNA. The RNA segments are encapsidated by the viral nucleocapsid (N) protein and form panhandle-like structures through interaction of complementary sequences at their 5' and 3' termini. Transcription and replication of a BUN genome analogue (minireplicon), comprising the viral non-coding sequences flanking a reporter gene, requires just the viral RNA polymerase (L protein) and N protein. Here, sequences of Bunyamwera serogroup M segment RNAs were compared and conserved elements within nt 20-33 of the 3' and 5' non-coding regions that can affect packaging of minireplicons into virions were identified. RNA-folding models suggest that a conserved sequence within nt 20-33 of the 5' end of the genome segments maintains conserved structural features necessary for efficient transcription. Competitive packaging experiments using M, L and S segment-derived minireplicons that encode different reporter genes showed variable packaging efficiencies of the three segments. Packaging of a particular segment appeared to be independent of the presence of other segments and, for the S segment, packaging efficiency was unaffected by the inclusion of viral coding sequences in the minireplicon. PMID- 16361431 TI - Identification of the Bunyamwera bunyavirus transcription termination signal. AB - Bunyamwera virus (BUNV) is the prototype of the family Bunyaviridae, which comprises segmented RNA viruses. Each of the BUNV negative-strand segments, small (S), medium (M) and large (L), serves as template for two distinct RNA-synthesis activities: (i) replication to generate antigenomes that are in turn replicated to yield further genomes; and (ii) transcription to generate a single species of mRNA. BUNV mRNAs are truncated at their 3' ends relative to the genome template, presumably because the BUNV transcriptase terminates transcription before reaching the 5' terminus of the genomic template. Here, identification of the transcription termination signal responsible for 3'-end truncation of BUNV S segment mRNA was carried out. It was shown that efficient transcription termination was signalled by a 33 nt sequence within the 5' non-translated region (NTR) of the S segment. A 6 nt region (3'-GUCGAC-5') within this sequence was found to play a major role in termination signalling, with other nucleotides possessing individually minor, but collectively significant, signalling ability. By abrogating the signalling ability of these 33 nt, we identified a second, functionally independent termination signal located 32 nt downstream. This downstream signal was 9 nt in length and contained a pentanucleotide sequence, 3' UGUCG-5', that overlapped the 6 nt major signalling component of the upstream signal. The pentanucleotide sequence was also found within the 5' NTR of the BUNV L segment and in several other members of the genus Orthobunyavirus, suggesting that the mechanism responsible for BUNV transcription termination may be common to other orthobunyaviruses. PMID- 16361432 TI - Liao ning virus, a new Chinese seadornavirus that replicates in transformed and embryonic mammalian cells. AB - Seadornaviruses are emerging arboviral pathogens from the south-east of Asia. The genus Seadornavirus contains two distinct species, Banna virus (BAV) isolated from humans with encephalitis and Kadipiro virus. BAV replicates within insect cells and mice but not in cultured mammalian cells. Here, the discovery of Liao ning virus (LNV), a new seadornavirus from the Aedes dorsalis mosquito, which was completely sequenced and was found to be related to BAV and Kadipiro virus, is reported. Two serotypes of LNV could be distinguished by a serum neutralization assay. According to amino acid identity with other seadornaviruses, and to criteria set by the ICTV for species delineation, LNV was identified as a member of a new species of virus. Its morphology was characterized by electron microscopy and found to be similar to that of BAV. LNV is the first reported seadornavirus that replicates in mammalian cells, leading to massive cytopathic effect in all transformed or embryonic cell lines tested. LNV- and BAV-infected mice producing a viraemia lasting for 5 days was followed by viral clearance. Mice infection generated virus quasi-species for LNV (the first reported observation for quasi-species in the family Reoviridae) but not for BAV. Challenge with BAV in mice immunized against BAV did not lead to productive infection. However, challenge with LNV in mice immunized against LNV was lethal with a new phase of viraemia and massive haemorrhage. PMID- 16361433 TI - Very virulent infectious bursal disease virus: reduced pathogenicity in a rare natural segment-B-reassorted isolate. AB - The purpose of this study was to compare the molecular epidemiology of infectious bursal disease virus (IBDV) segments A and B of 50 natural or vaccine IBDV strains that were isolated or produced between 1972 and 2002 in 17 countries from four continents, with phenotypes ranging from attenuated to very virulent (vv). These strains were subjected to sequence and phylogenetic analysis based on partial sequences of genome segments A and B. Although there is co-evolution of the two genome segments (70 % of strains kept the same genetic relatives in the segment A- and B-defined consensus trees), several strains (26 %) were identified with the incongruence length difference test as exhibiting a significantly different phylogenetic relationship depending on which segment was analysed. This suggested that natural reassortment could have occurred. One of the possible naturally occurring reassortant strains, which exhibited a segment A related to the vvIBDV cluster whereas its segment B was not, was thoroughly sequenced (coding sequence of both segments) and submitted to a standardized experimental characterization of its acute pathogenicity. This strain induced significantly less mortality than typical vvIBDVs; however, the mechanisms for this reduced pathogenicity remain unknown, as no significant difference in the bursal lesions, post-infectious antibody response or virus production in the bursa was observed in challenged chickens. PMID- 16361434 TI - Identification of B-cell epitopes in the capsid protein of avian hepatitis E virus (avian HEV) that are common to human and swine HEVs or unique to avian HEV. AB - Avian hepatitis E virus (avian HEV) was recently discovered in chickens from the USA that had hepatitis-splenomegaly (HS) syndrome. The complete genomic sequence of avian HEV shares about 50 % nucleotide sequence identity with those of human and swine HEVs. The open reading frame 2 (ORF2) protein of avian HEV has been shown to cross-react with human and swine HEV ORF2 proteins, but the B-cell epitopes in the avian HEV ORF2 protein have not been identified. Nine synthetic peptides from the predicted four antigenic domains of the avian HEV ORF2 protein were synthesized and corresponding rabbit anti-peptide antisera were generated. Using recombinant ORF2 proteins, convalescent pig and chicken antisera, peptides and anti-peptide rabbit sera, at least one epitope at the C terminus of domain II (possibly between aa 477-492) that is unique to avian HEV, one epitope in domain I (aa 389-410) that is common to avian, human and swine HEVs, and one or more epitopes in domain IV (aa 583-600) that are shared between avian and human HEVs were identified. Despite the sequence difference in ORF2 proteins between avian and mammalian HEVs and similar ORF2 sequence between human and swine HEV ORF2 proteins, rabbit antiserum against peptide 6 (aa 389-399) recognized only human HEV ORF2 protein, suggesting complexity of the ORF2 antigenicity. The identification of these B-cell epitopes in avian HEV ORF2 protein may be useful for vaccine design and may lead to future development of immunoassays for differential diagnosis of avian, swine and human HEV infections. PMID- 16361435 TI - Importance of the C-terminal domain of soybean mosaic virus coat protein for subunit interactions. AB - The potyvirus coat protein (CP) is involved in aphid transmission, cell-to-cell movement and virus assembly, not only by binding to viral RNA, but also by self interaction or interactions with other factors. In this study, a number of CP mutants of Soybean mosaic virus (SMV) containing deletions and site-directed mutations were generated and cloned into yeast two-hybrid vectors. Interaction was confirmed by the expression of reporter genes, including HIS3, ADE2 and MEL1, in yeast strain AH109. Deletion of the C-terminal region of the CP caused loss of the CP-CP self-interaction ability detected in CP mutants with the C-terminal region. Alanine substitution at the amino acid positions R190, E191, E212, R245, H246 and R249 disrupted CP-CP interaction, whereas substitutions at the amino acid positions R188, D189, D198, K205, K218 and D250 did not. These results indicate that the C-terminal region of SMV CP may contain a domain(s) or amino acids required for CP-CP interaction and virus assembly. PMID- 16361436 TI - Variants of Peach latent mosaic viroid inducing peach calico: uneven distribution in infected plants and requirements of the insertion containing the pathogenicity determinant. AB - Previous characterization of Peach latent mosaic viroid (PLMVd) variants from a single peach calico (PC) isolate showed that PC symptoms are induced by variants with a 12-13 nt insertion at a specific position and folding into a hairpin with a U-rich loop. Here, this study was extended to two other PC isolates. PLMVd variants with insertions similar to those reported previously (type 1), predominated in one isolate (PC-P2). The second (PC-P1), in addition to these variants, contained others with insertions in the same position and of the same size, but with the hairpin capped by a GA-rich loop (type 2). When symptomatic and non-symptomatic tissues from both isolates were used to inoculate GF-305 peach seedlings, they reproduced the phenotype of the inoculum source, indicating that variants differing in pathogenicity are unevenly distributed within single plants. Moreover, characterization of the progeny from inoculations with the PC P1 source showed that variants with insertions of type 1 and 2 were predominant in the symptomatic and non-symptomatic seedlings, respectively, confirming the association between PC and variants with type 1 but not type 2 insertions. Inoculations with dimeric in vitro transcripts from PLMVd variants with type 1, type 2 and with a chimeric insertion showed that the variant with type 2 insertion was latent and established that the U-rich capping loop has a major role in PC, although the adjacent stem may also have some influence. Insertions can be acquired and lost during infection, suggesting that latent variants can evolve into pathogenic variants and vice versa. PMID- 16361437 TI - Characterization of debilitation-associated mycovirus infecting the plant pathogenic fungus Sclerotinia sclerotiorum. AB - It was previously reported that three dsRNA segments, designated L, M and S, were isolated from Sclerotinia sclerotiorum strain Ep-1PN and that the M dsRNA segment was coincident with hypovirulence and debilitation of the fungal host. Here, the complete nucleotide sequence of the M dsRNA of 5419 nt, excluding the poly(A) tail, was determined. Sequence analysis revealed the occurrence of a single open reading frame (nt 93-5195) encoding a protein with significant similarity to the replicases of the 'alphavirus-like' supergroup of positive-strand RNA viruses. The M dsRNA-encoded putative replicase protein contained the conserved methyl transferase, helicase and RNA-dependent RNA polymerase (RdRp) domains characteristic of the replicases of potex-like plant viruses (flexiviruses) and Botrytis virus F (BVF), a flexuous rod mycovirus infecting the phytopathogenic fungus Botrytis cinerea. Furthermore, convincing evidence is presented showing that ascospore descendents derived from the debilitated strain Ep-1PN were devoid of dsRNA and exhibited normal colony morphology. Moreover, it was demonstrated that the debilitation phenotype was transmitted from the parental debilitated strain to its normal ascospore progeny via hyphal anastomosis. These results suggest that the M dsRNA from strain Ep-1PN is derived from the genomic RNA of a positive-strand RNA virus, which we designated Sclerotinia sclerotiorum debilitation-associated RNA virus (SsDRV). Although phylogenetic analysis of the conserved RdRp motifs verified that SsDRV is closely related to BVF and to the allexiviruses in the family Flexiviridae, SsDRV is distinct from these viruses, mainly based on the lack of coat protein and movement protein. PMID- 16361438 TI - Pathological prion protein in muscles of hamsters and mice infected with rodent adapted BSE or vCJD. AB - Recently, pathological prion protein (PrP(TSE)) was detected in muscle from sheep infected with scrapie, the archetype of transmissible spongiform encephalopathies (TSEs). This finding has highlighted the question of whether mammalian muscle may potentially also provide a reservoir for TSE agents related to bovine spongiform encephalopathy (BSE) and variant Creutzfeldt-Jakob Disease (vCJD). Here, results are reported from studies in hamsters and mice that provide direct experimental evidence, for the first time, of BSE- and vCJD-associated PrP(TSE) deposition in muscles. Our findings emphasize the need for further assessment of possible public-health risks from TSE involvement of skeletal muscle. PMID- 16361439 TI - Network of dendritic cells within the muscular layer of the mouse intestine. AB - Dendritic cells (DCs) are located at body surfaces such as the skin, respiratory and genital tracts, and intestine. To further analyze intestinal DCs, we adapted an epidermal sheet separation technique and obtained two intestinal layers, facing the lumen and serosa. Unexpectedly, immunolabeling of the layer toward the serosa revealed a regular, dense, planar network of cells with prominent dendritic morphology within the external muscular layer and with increasing frequency along the length of the intestine. Direct examination of the serosal disposed layers showed a significant fraction of the DCs to express DEC 205/CD205, CD11c, Langerin/CD207, Fcgamma receptor/CD16/32, CD14, and low levels of activation markers, CD25, CD80, CD86, and CD95. By more sensitive FACS analyses, cells from this layer contained two CD11c(+) populations of CD45(+) CD205(+), CD19(-) leukocytes, MHC II(+) and MHC II(-). When ovalbumin conjugated to an anti-DEC-205 antibody was injected into mice, the conjugate targeted to these DCs, which upon isolation were able to stimulate ovalbumin-specific, CD4(+) and CD8(+) T cell antigen receptor-transgenic T cells. In vivo, these DCs responded to two microbial stimuli, systemic LPS and oral live bacteria, by up regulating CD80, CD86, DEC-205, and Langerin within 12 h. This network of DCs thus represents a previously unrecognized antigen-presenting cell system in the intestine. PMID- 16361440 TI - A cancer DNA phenotype in healthy prostates, conserved in tumors and adjacent normal cells, implies a relationship to carcinogenesis. AB - A cancer DNA phenotype, identical to the DNA structure of tumors, has been identified in the prostate glands of certain healthy men over 55 years of age. We now show that the same DNA signature exists in normal tissues adjacent to tumors. This finding implies that the phenotype is maintained in normal prostate cells from its inception through tumor development. The presence of the phenotype in tumors, adjacent normal cells, and in the normal prostate cells of certain older men suggests that it is a potentially critical factor in tumor development and may serve as an early biomarker for cancer risk assessment. Intervention to inhibit the development of the phenotype in healthy men, or to eliminate it once formed, may suppress or even prevent tumor formation. PMID- 16361441 TI - Absolute rate theories of epigenetic stability. AB - Spontaneous switching events in most characterized genetic switches are rare, resulting in extremely stable epigenetic properties. We show how simple arguments lead to theories of the rate of such events much like the absolute rate theory of chemical reactions corrected by a transmission factor. Both the probability of the rare cellular states that allow epigenetic escape and the transmission factor depend on the rates of DNA binding and unbinding events and on the rates of protein synthesis and degradation. Different mechanisms of escape from the stable attractors occur in the nonadiabatic, weakly adiabatic, and strictly adiabatic regimes, characterized by the relative values of those input rates. PMID- 16361442 TI - Up-regulation of a H+-pyrophosphatase (H+-PPase) as a strategy to engineer drought-resistant crop plants. AB - Engineering drought -resistant crop plants is a critically important objective. Overexpression of the vacuolar H(+)-pyrophosphatase (H(+)-PPase) AVP1 in the model plant Arabidopsis thaliana results in enhanced performance under soil water deficits. Recent work demonstrates that AVP1 plays an important role in root development through the facilitation of auxin fluxes. With the objective of improving crop performance, we expressed AVP1 in a commercial cultivar of tomato. This approach resulted in (i) greater pyrophosphate-driven cation transport into root vacuolar fractions, (ii) increased root biomass, and (iii) enhanced recovery of plants from an episode of soil water deficit stress. More robust root systems allowed transgenic tomato plants to take up greater amounts of water during the imposed water deficit stress, resulting in a more favorable plant water status and less injury. This study documents a general strategy for improving drought resistance of crops. PMID- 16361443 TI - Structural basis for conductance by the archaeal aquaporin AqpM at 1.68 A. AB - To explore the structural basis of the unique selectivity spectrum and conductance of the transmembrane channel protein AqpM from the archaeon Methanothermobacter marburgensis, we determined the structure of AqpM to 1.68-A resolution by x-ray crystallography. The structure establishes AqpM as being in a unique subdivision between the two major subdivisions of aquaporins, the water selective aquaporins, and the water-plus-glycerol-conducting aquaglyceroporins. In AqpM, isoleucine replaces a key histidine residue found in the lumen of water channels, which becomes a glycine residue in aquaglyceroporins. As a result of this and other side-chain substituents in the walls of the channel, the channel is intermediate in size and exhibits differentially tuned electrostatics when compared with the other subfamilies. PMID- 16361444 TI - Mineralocorticoid receptors are indispensable for nongenomic modulation of hippocampal glutamate transmission by corticosterone. AB - The adrenal hormone corticosterone transcriptionally regulates responsive genes in the rodent hippocampus through nuclear mineralocorticoid and glucocorticoid receptors. Via this genomic pathway the hormone alters properties of hippocampal cells slowly and for a prolonged period. Here we report that corticosterone also rapidly and reversibly changes hippocampal signaling. Stress levels of the hormone enhance the frequency of miniature excitatory postsynaptic potentials in CA1 pyramidal neurons and reduce paired-pulse facilitation, pointing to a hormone dependent enhancement of glutamate-release probability. The rapid effect by corticosterone is accomplished through a nongenomic pathway involving membrane located receptors. Unexpectedly, the rapid effect critically depends on the classical mineralocorticoid receptor, as evidenced by the effectiveness of agonists, antagonists, and brain-specific inactivation of the mineralocorticoid but not the glucocorticoid receptor gene. Rapid actions by corticosterone would allow the brain to change its function within minutes after stress-induced elevations of corticosteroid levels, in addition to responding later through gene mediated signaling pathways. PMID- 16361446 TI - Effectiveness of primary health care evaluated by a longitudinal ecological approach. PMID- 16361445 TI - Disruption of Cryptochrome partially restores circadian rhythmicity to the arrhythmic period mutant of Drosophila. AB - The Drosophila melanogaster circadian clock is generated by interlocked feedback loops, and null mutations in core genes such as period and timeless generate behavioral arrhythmicity in constant darkness. In light-dark cycles, the elevation in locomotor activity that usually anticipates the light on or off signals is severely compromised in these mutants. Light transduction pathways mediated by the rhodopsins and the dedicated circadian blue light photoreceptor cryptochrome are also critical in providing the circadian clock with entraining light signals from the environment. The cry(b) mutation reduces the light sensitivity of the fly's clock, yet locomotor activity rhythms in constant darkness or light-dark cycles are relatively normal, because the rhodopsins compensate for the lack of cryptochrome function. Remarkably, when we combined a period-null mutation with cry(b), circadian rhythmicity in locomotor behavior in light-dark cycles, as measured by a number of different criteria, was restored. This effect was significantly reduced in timeless-null mutant backgrounds. Circadian rhythmicity in constant darkness was not restored, and TIM protein did not exhibit oscillations in level or localize to the nuclei of brain neurons known to be essential for circadian locomotor activity. Therefore, we have uncovered residual rhythmicity in the absence of period gene function that may be mediated by a previously undescribed period-independent role for timeless in the Drosophila circadian pacemaker. Although we do not yet have a molecular correlate for these apparently iconoclastic observations, we provide a systems explanation for these results based on differential sensitivities of subsets of circadian pacemaker neurons to light. PMID- 16361447 TI - Health promotion evaluation and the principle of prevention. PMID- 16361448 TI - Indicators of socioeconomic position (part 1). AB - This glossary presents a comprehensive list of indicators of socioeconomic position used in health research. A description of what they intend to measure is given together with how data are elicited and the advantages and limitation of the indicators. The glossary is divided into two parts for journal publication but the intention is that it should be used as one piece. The second part highlights a life course approach and will be published in the next issue of the journal. PMID- 16361449 TI - Evaluation of the impact of the Family Health Program on infant mortality in Brazil, 1990-2002. AB - OBJECTIVE: To use publicly available secondary data to assess the impact of Brazil's Family Health Program on state level infant mortality rates (IMR) during the 1990s. DESIGN: Longitudinal ecological analysis using panel data from secondary sources. Analyses controlled for state level measures of access to clean water and sanitation, average income, women's literacy and fertility, physicians and nurses per 10,000 population, and hospital beds per 1,000 population. Additional analyses controlled for immunisation coverage and tested interactions between Family Health Program and proportionate mortality from diarrhoea and acute respiratory infections. SETTING: 13 years (1990-2002) of data from 27 Brazilian states. MAIN RESULTS: From 1990 to 2002 IMR declined from 49.7 to 28.9 per 1,000 live births. During the same period average Family Health Program coverage increased from 0% to 36%. A 10% increase in Family Health Program coverage was associated with a 4.5% decrease in IMR, controlling for all other health determinants (p<0.01). Access to clean water and hospital beds per 1,000 were negatively associated with IMR, while female illiteracy, fertility rates, and mean income were positively associated with IMR. Examination of interactions between Family Health Program coverage and diarrhoea deaths suggests the programme may reduce IMR at least partly through reductions in diarrhoea deaths. Interactions with deaths from acute respiratory infections were ambiguous. CONCLUSIONS: The Family Health Program is associated with reduced IMR, suggesting it is an important, although not unique, contributor to declining infant mortality in Brazil. Existing secondary datasets provide an important tool for evaluation of the effectiveness of health services in Brazil. PMID- 16361450 TI - Disease profile of children in Kabul: the unmet need for health care. AB - This study was conducted at the paediatric emergency department of a tertiary care teaching and referral hospital in Kabul, Afghanistan to assess the morbidity and mortality pattern of illness in paediatric population. Afghanistan has one of the highest infant mortality rates in the world, and there is complete breakdown of the health care system in the country because of the continued war. A total of 17,850 children and neonates were seen at the paediatric emergency centre at IGICH, Kabul in one year period from 18 September 2002 to 17 September 2003. The most common illnesses were diarrhoea and respiratory infections. Infectious diseases, neonatal illnesses, and cardiac diseases were other important causes of morbidity. Neonatal deaths formed the major proportion of all deaths. Morbidity and mortality attributable to easily preventable/curable diseases was quite high. There is an urgent need to develop an integrated and effective health care system in the country. PMID- 16361451 TI - Economic efficiency of gate-keeping compared with fee for service plans: a Swiss example. AB - STUDY OBJECTIVE: The impact of isolated gate-keeping on health care costs remains unclear. The aim of this study was to assess to what extent lower costs in a gate keeping plan compared with a fee for service plan were attributable to more efficient resource management, or explained by risk selection. DESIGN: Year 2000 costs to the Swiss statutory sick funds and potentially relevant covariates were assessed retrospectively from beneficiaries participating in an observational study, their primary care physicians, and insurance companies. To adjust for case mix, two-part regression models of health care costs were fitted, consisting of logistic models of any costs occurring, and of generalised linear models of the amount of costs in persons with non-zero costs. Complementary data sources were used to identify selection effects. SETTING: A gate-keeping plan introduced in 1997 and a fee for service plan, in Aarau, Switzerland. PARTICIPANTS: Of each plan, 905 randomly selected adult beneficiaries were invited. The overall participation rate was 39%, but was unevenly distributed between plans. MAIN RESULTS: The characteristics of gate-keeping and fee for service beneficiaries were largely similar. Unadjusted total costs per person were Sw fr 231 (8%) lower in the gate-keeping group. After multivariate adjustment, the estimated cost savings achieved by replacing fee for service based health insurance with gate keeping in the source population amounted to Sw fr 403-517 (15%-19%) per person. Some selection effects were detected but did not substantially influence this result. An impact of non-detected selection effects cannot be ruled out. CONCLUSIONS: This study hints at substantial cost savings through gate-keeping that are not attributable to mere risk selection. PMID- 16361452 TI - Social class inequalities in perinatal outcomes: Scotland 1980-2000. AB - OBJECTIVE: To examine social class inequalities in adverse perinatal events in Scotland between 1980 and 2000 and how these were influenced by other maternal risk factors. DESIGN: Population based study using routine maternity discharge data. SETTING: Scotland. PARTICIPANTS: All women who gave birth to a live singleton baby in Scottish hospitals between 1980 and 2000 (n=1,282,172). MAIN OUTCOME MEASURES: Low birth weight (LBW), preterm birth, and small for gestational age (SGA). RESULTS: The distribution of social class changed over time, with the proportion of mothers with undetermined social class increasing from 3.9% in 1980-84 to 14.8% in 1995-2000. The relative index of inequality (RII) decreased during the 1980s for all outcomes. The RII then increased between the early and late 1990s (LBW from 2.09 (95%CI 1.97, 2.22) to 2.43 (2.29, 2.58), preterm from 1.52 (1.44, 1.61) to 1.75 (1.65, 1.86), and SGA from 2.28 (2.14, 2.42) to 2.49 (2.34, 2.66) respectively). Inequalities were greatest in married mothers, mothers aged over 35, mothers taller than 164 cm, and mothers with a parity of one or more. Inequalities were also greater by the end of the 1990s than at the start of the 1980s for women of parity one or more and for mothers who were not married. CONCLUSION: Despite decreasing during the 1980s, inequalities in adverse perinatal outcomes increased during the 1990s in all strata defined by maternal characteristics. PMID- 16361453 TI - Associations of area based deprivation status and individual educational attainment with incidence, treatment, and prognosis of first coronary event in Rome, Italy. AB - BACKGROUND: Socioeconomic gradients in the occurrence of myocardial infarction are well known, but few studies have examined socioeconomic disparities in post infarction outcomes. The objective of this study was to explore relations of socioeconomic status with the incidence, treatment, and outcome of first coronary event in Rome, Italy, during the period 1998-2000, examining effect modification by gender. METHODS: Subjects were Rome residents aged 35-84 years who died from first acute coronary event before reaching the hospital (n=3470) or were hospitalised for first acute myocardial infarction (n=8467). Area based deprivation status and patients' educational attainment were the exposure variables. The outcomes were: incidence of coronary event; recanalisation at the index hospitalisation and fatality within 28 days of hospitalisation; cardiac readmissions and fatality between 28 days and one year of index hospitalisation. RESULTS: Incidence rates increased as area based deprivation status increased; the effect was stronger among women than among men (men RR=1.40, 95%CI:1.30, 1.50, women RR=1.78, 95%CI:1.60, 1.98, most compared with least deprived). Rates of recanalisation were significantly lower in the most deprived patients than in the least deprived (OR=0.77, 95%CI:0.59, 0.99) and in the less educated than in the highly educated (OR=0.73, 95%CI:0.58, 0.90). Associations of short term fatality with area based deprivation status and educational attainment were weak and inconsistent. However, neither deprivation status nor education was associated with one year outcomes. CONCLUSIONS: Area based deprivation status is strongly related to incidence of coronary events, and more so among women than among men. Deprivation status and educational attainment are weakly and inconsistently associated with short term fatality but seem not to influence one year prognosis of acute myocardial infarction. Deprived and less educated patients experience limited access to recanalisation procedures. PMID- 16361454 TI - Job control, job demands, or social class? The impact of working conditions on the relation between social class and health. AB - BACKGROUND: The aim of the study was to investigate (1) how much of the association between health and social class is accounted by psychosocial working conditions, and (2) whether health is related to working conditions after controlling for social class. METHODS: The data derive from the surveys of the Helsinki health study, collected in 2000, 2001, and 2002 from 40-60 year old employees working for the City of Helsinki (n=8970, response rate 67%). The study measured occupation based social class and Karasek's demand-control model. The health outcomes were self rated health as less than good and limiting longstanding illness. Age adjusted prevalence percentages and fitted logistic regression models were calculated. RESULTS: The individual effects of social class and psychosocial working conditions on self rated health and limiting longstanding illness were strong among both men and women. The relation between social class and both health outcomes considerably attenuated when job control was controlled for, but was reinforced when controlling for job demands. Controlling for both job control and job demands attenuated the relation between social class and self rated health and limiting longstanding illness among women, however, was reinforced among men. CONCLUSIONS: A substantial part of the relation between social class and health could be attributed to job control, however, job demands reinforced the relation. Although the effect of social class is mediated by psychosocial working conditions, both social class and working conditions were related to health after mutual adjustments. PMID- 16361455 TI - Influence of change in psychosocial work characteristics on sickness absence: The Whitehall II Study. AB - OBJECTIVES: To study the influence of change in self perceived psychosocial work characteristics on subsequent rates of sickness absence. METHODS: Prospective cohort study of British civil service employees. Job control, job demands, and work social supports were measured in 1985/88 and in 1991/93. Analyses included 3817 British civil servants with sickness absence records at baseline (1985-89) and for two follow up periods, early (1994-95) and later follow up (1996-98). RESULTS: Change in work characteristics predicted subsequent incidence of long spells of sickness absence (>7 days) in the early follow up period after adjustment for covariates including baseline work characteristics, health status, and sickness absence. Adjusted rate ratios were 1.23 (95% CI 1.03 to 1.46) for decreased compared with stable decision latitude; 1.17 (95% CI 1.01 to 1.36) for increased compared with stable job demands and 0.79 (95% CI 0.67 to 0.93) for increased compared with stable work social support. These associations were also seen in a sub-sample who did not change employment grade. In the later follow up period, associations between work change and long spells of sickness absence were similar for decision latitude, less pronounced for job demands, and no longer apparent for social supports. Changes in work characteristics were not associated with subsequent short spells of sickness absence ( 0 and assuming equal priors, these probabilities were > 99.5% in 10 of 12 possible breed contrasts. For the 2 most closely related breeds (FST = 0.041), this probability was 96.3%, and the probability of excluding the origin of an animal from an alleged breed when it was actually derived from another breed was similar. PMID- 16361487 TI - Success at first insemination in Australian Angus cattle: analysis of uncertain binary responses. AB - Field data from Australian Angus herds were used to investigate 2 methods of analyzing uncertain binary responses for success or failure at first insemination. A linear mixed model that included herd, year, and month of mating as fixed effects; unrelated service sire, additive animal, and residual as random effects; and linear and quadratic effects of age at mating as covariates was used to analyze binary data. An average gestation length (GL) derived from artificial insemination data was used to assign an insemination date to females mated to natural service sires. Females that deviated from this average GL led to uncertain binary responses. Two analyses were carried out: 1) a threshold model fitted to uncertain binary data, ignoring uncertainty (M1); and 2) a threshold model fitted to uncertain binary data, accounting for uncertainty via fuzzy logic classification (M2). There was practically no difference between point estimates obtained from M1 and M2 for service sire and herd variance; however, when uncertain binary data were analyzed ignoring uncertainty (M1), additive variance and heritability estimates were greater than with M2. Pearson correlations indicated that no major reranking would be expected for service sire effects and animal breeding values using M1 and M2. Given the results of the current study, a threshold model contemplating uncertainty is suggested for noisy binary data to avoid bias when estimating genetic parameters. PMID- 16361488 TI - Age of dam and sex of calf adjustments and genetic parameters for gestation length in Charolais cattle. AB - To estimate adjustment factors and genetic parameters for gestation length (GES), AI and calving date records (n = 40,356) were extracted from the Canadian Charolais Association field database. The average time from AI to calving date was 285.2 d (SD = 4.49 d) and ranged from 274 to 296 d. Fixed effects were sex of calf, age of dam (2, 3, 4, 5 to 10, > or = 11 yr), and gestation contemporary group (year of birth x herd of origin). Variance components were estimated using REML and 4 animal models (n = 84,332) containing from 0 to 3 random maternal effects. Model 1 (M1) contained only direct genetic effects. Model 2 (M2) was G1 plus maternal genetic effects with the direct x maternal genetic covariance constrained to zero, and model 3 (M3) was G2 without the covariance constraint. Model 4 (M4) extended G3 to include a random maternal permanent environmental effect. Direct heritability estimates were high and similar among all models (0.61 to 0.64), and maternal heritability estimates were low, ranging from 0.01 (M2) to 0.09 (M3). Likelihood ratio tests and parameter estimates suggested that M4 was the most appropriate (P < 0.05) model. With M4, phenotypic variance (18.35 d2) was partitioned into direct and maternal genetic, and maternal permanent environmental components (hd2 = 0.64 +/- 0.04, hm2 = 0.07 +/- 0.01, r(d,m) = 0.37 +/- 0.06, and c2 = 0.03 +/- 0.01, respectively). Linear contrasts were used to estimate that bull calves gestated 1.26 d longer (P < 0.02) than heifers, and adjustments to a mature equivalent (5 to 10 yr old) age of dam were 1.49 (P < 0.01), 0.56 (P < 0.01), 0.33 (P < 0.01), and -0.24 (P < 0.14) d for GES records of calves born to 2-, 3-, 4-, and > or = 11-yr-old cows, respectively. Bivariate animal models were used to estimate genetic parameters for GES with birth and adjusted 205-d weaning weights, and postweaning gain. Direct GES was positively correlated with direct birth weight (BWT; 0.34 +/- 0.04) but negatively correlated with maternal BWT (-0.20 +/- 0.07). Maternal GES had a low, negative genetic correlation with direct BWT (-0.15 +/- 0.05) but a high and positive genetic correlation with maternal BWT (0.62 +/- 0.07). Generally, GES had near zero genetic correlations with direct and maternal weaning weights. Results suggest that important genetic associations exist for GES with BWT, but genetic correlations with weaning weight and postweaning gain were less important. PMID- 16361489 TI - Genetic and phenotypic relationships of farrowing and weaning survival to birth and placental weights in pigs. AB - Data obtained during 4 generations of divergent selection for placental efficiency were used to determine factors influencing survival at farrowing and weaning in litters produced by first-parity females. Data were collected from 193 litters and included records on 2,053 individuals. Farrowing survival (FS) and weaning survival (WS) were considered traits of the piglet and were scored 1 if the individual was alive at a time point or 0 if dead. Estimates of (co)variance components for direct and maternal additive genetic effects for FS and WS were obtained using an animal model and computed with the MTDFREML program. Estimates of direct heritability were 0.16 for FS and 0.18 for WS. Estimates of maternal heritability were 0.14 for FS and 0.10 for WS. Genetic correlation estimates between direct and maternal effects were high and negative for both traits. The direct genetic correlation between FS and WS was 0.92. Variables associated with FS and WS were determined using logistic regression procedures. Birth weight (BRW), placental weight, their interaction, and total born can be used as predictors of survival at farrowing in the absence of estimates of genetic merit for survival. The same model, excluding total number born, was the best model for predicting WS. In the presence of BRW information, placental efficiency did not improve the prediction of survival. While it was clearly disadvantageous for a piglet to be below the litter mean in BRW, being above the mean did not provide a substantial advantage in survival. Results from this analysis suggest that it is possible to select for increased survival at farrowing and at weaning. Information on a piglet's BRW, placental weight, litter average BRW, and deviation from litter average BRW can be used to optimize those values at levels resulting in high survival probability. PMID- 16361490 TI - Factors associated with ELISA scores for paratuberculosis in an Angus-Brahman multibreed herd of beef cattle. AB - Cow and calf genetic and environmental factors were evaluated for their association with ELISA scores for paratuberculosis in a multibreed population of beef cattle. The ELISA scores are a measure of the presence or absence of antibodies against Mycobacterium avium subsp. paratuberculosis in bovine serum. The linear mixed-model analysis used 352 ELISA scores from 238 cows: 51 Angus (A); 34 Brahman (B); 41 (3/4 A 1/4 B); 45 (1/2 A 1/2 B); 34 (1/4 A 3/4 B); and 33 Brangus (5/8 A 3/8 B). Cows were assumed to be unrelated. Year affected (P < 0.001) ELISA scores, but age of cow did not, which was expected to be significant because of the chronic progressive nature of this disease. Important regressions on fixed effects associated with cows were 1) a positive estimate of cow B breed effect (0.59 +/- 0.24; P < 0.017), indicating an upward trend of ELISA scores toward 100% B cows; 2) a negative estimate for weight change from before calving (late November) to the date of the blood sample in May (-0.0062 +/- 0.0019 score/kg; P < 0.002), indicating that poorer maintenance of cow weights was associated with higher ELISA scores; and 3) a positive estimate for days in lactation of cow on the date of the blood sample (0.0086 +/- 0.0034 score/d; P < 0.021), indicating the production of larger amounts of antibodies against Mycobacterium avium subsp. paratuberculosis as lactation progressed. Relevant regressions on fixed effects associated with calves were 1) calf birth weight ( 0.022 +/- 0.010 score/kg; P < 0.035), and 2) calf gain from birth to the date of the cow blood sample (-0.0092 +/- 0.0027 score/kg; P < 0.001). These estimates indicate that cows that produced lighter calves at birth and/or calves with slower preweaning growth tended to have greater ELISA scores. Although the sensitivity (percentage of infected animals detected) of ELISA was only 50%, these results suggest that subclinical paratuberculosis may be negatively affecting cows and their offspring. Factors identified as associated with ELISA scores could help producers with culling decisions related to paratuberculosis control and eradication in beef cattle. PMID- 16361491 TI - An investigation of susceptibility to porcine reproductive and respiratory syndrome virus between two genetically diverse commercial lines of pigs. AB - The objective of this study was to determine whether host genetics play a role in susceptibility to the respiratory disease in growing pigs caused by the porcine reproductive and respiratory syndrome virus (PRRSV). Based on a previous study, 2 genetically diverse commercial lines of pigs that also were divergent in the susceptibility of monocyte-derived macrophages to PRRSV infection in vitro were selected for an in vivo challenge study. Based on the average percentage of infected macrophages for each line, a line derived from the Large White breed was characterized as fluorescence-activated cell sorting(hi) (FACS(hi)), and a line derived from Duroc and Pietrain breeds was characterized as FACS(lo). Pigs from each line were challenged at 6 wk of age with PRRSV VR-2385 and necropsied at 10 or 21 d after infection. Data collected included clinical evaluation of disease, virus titration in serum and lung lavage fluid, macroscopic lung lesion scores, and microscopic lung lesion scores. The FACS(lo) line had consistently more severe clinical disease compared with the FACS(hi) line in the early stages of infection. Differences between line means were significant (P < 0.05) at 10 d after infection for all variables just described, and the FACS(lo) line showed more severe signs of disease. By 21 d after infection, clinical signs and lesions were resolving, and the differences between lines were significant (P < 0.04) only for microscopic lung lesion scores but approached significance (P < 0.08) for virus titer in serum. At 21 d after infection, the relationship between the lines reversed; the FACS(hi) line had higher serum virus titers than the FACS(lo) line. This report provides evidence that strongly suggests the existence of a host genetic component in disease susceptibility to PRRSV and indicates that further study is warranted to define the cellular mechanisms that affect disease susceptibility. PMID- 16361492 TI - Crossbreeding parameter estimation for functional longevity in rabbits using survival analysis methodology. AB - A complete diallel cross involving 3 maternal lines of rabbit was performed to estimate cross-breeding parameters for functional longevity. This trait was defined as the ability to delay involuntary culling. The lines considered, A, V, and Prat, had all been selected by litter size at weaning for a long period. Data were related to a total of 653 does belonging to the 9 genetic types from the diallel cross; does were reared and bred on the same commercial farm. Survival analysis was performed using a Cox proportional hazard model. The model incorporated time-dependent factors, such as year-season, litter size, and the interaction between cycle and physiological status of the female; time independent factors, such as the genetic type of the doe; and sire and dam random factors. Crossbreeding parameters were estimated from the solutions obtained for the type of doe and its estimated variance-covariance matrix, using a generalized least squares procedure. The estimated parameters were the differences between lines in direct genetic effects and maternal genetic effects and individual heterosis. Relevant differences were observed in direct genetic effects between lines A and Prat but not in any maternal effects. Heterosis was found to be significant and favorable between lines A and Prat, and between the lines V and Prat. The magnitude of this effect was variable but important, especially in the first cross. Results stress the importance of using crosses between specialized lines to produce does for intensive meat rabbit production. PMID- 16361493 TI - Effects of age, weight, and fat slaughter end points on estimates of breed and retained heterosis effects for carcass traits. AB - The influence of different levels of adjusted fat thickness (AFT) and HCW slaughter end points (covariates) on estimates of breed and retained heterosis effects was studied for 14 carcass traits from serially slaughtered purebred and composite steers from the US Meat Animal Research Center (MARC). Contrasts among breed solutions were estimated at 0.7, 1.1, and 1.5 cm of AFT, and at 295.1, 340.5, and 385.9 kg of HCW. For constant slaughter age, contrasts were adjusted to the overall mean (432.5 d). Breed effects for Red Poll, Hereford, Limousin, Braunvieh, Pinzgauer, Gelbvieh, Simmental, Charolais, MARC I, MARC II, and MARC III were estimated as deviations from Angus. In addition, purebreds were pooled into 3 groups based on lean-to-fat ratio, and then differences were estimated among groups. Retention of combined individual and maternal heterosis was estimated for each composite. Mean retained heterosis for the 3 composites also was estimated. Breed rankings and expression of heterosis varied within and among end points. For example, Charolais had greater (P < 0.05) dressing percentages than Angus at the 2 largest levels of AFT and smaller (P < 0.01) percentages at the 2 largest levels of HCW, whereas the 2 breeds did not differ (P > or = 0.05) at a constant age. The MARC III composite produced 9.7 kg more (P < 0.01) fat than Angus at AFT of 0.7 cm, but 7.9 kg less (P < 0.05) at AFT of 1.5 cm. For MARC III, the estimate of retained heterosis for HCW was significant (P < 0.05) at the lowest level of AFT, but at the intermediate and greatest levels estimates were nil. The pattern was the same for MARC I and MARC III for LM area. Adjustment for age resulted in near zero estimates of retained heterosis for AFT, and similarly, adjustment for HCW resulted in nil estimates of retained heterosis for LM area. For actual retail product as a percentage of HCW, the estimate of retained heterosis for MARC III was negative (-1.27%; P < 0.05) at 0.7 cm but was significantly positive (2.55%; P < 0.05) at 1.5 cm of AFT. Furthermore, for MARC III, estimates of heterosis for some traits (fat as a percentage of HCW as another example) also doubled in magnitude depending on different levels of AFT end point. Rational exploitation of breeds requires special attention to use of different end points and levels of those end points, mainly for fat thickness. PMID- 16361494 TI - Predicting breeding values and accuracies from group in comparison to individual observations. AB - Individual observations are routinely used in livestock evaluations. In some cases, pooled data representing the joint but not individual performance of a group of animals may be available. For example, pooled feed intake may be measured on a pen of livestock. The usual mixed model approach to genetic evaluation can still be applied as an exact method in this setting, provided incidence and residual variance-covariance matrices are suitably modified to account for the pooling. Approximate evaluations may be achieved by treating average performance as if it pertained to each individual in the pool. Theoretical accuracies can be obtained as a function of elements of the inverse coefficient matrix. A 3-generation data set representing 1,000 animals with feed intake observations from 49 sires and 200 maternal grand sires was simulated with heritability of 0.34. Individual records were pooled to represent circumstances in which animals with records were collectively measured. Animals were allocated into pens at random, by sire, or by maternal grand sire. Simulation was replicated with unique fixed effects for each pen. Following evaluation from each method, the empirical accuracy or product-moment correlation between true (simulated) and estimated merit could be quantified. The analysis of individual observations resulted in empirical accuracy of 0.63 for animals on test and 0.77 for their sires. Pooling the observations in pens of 2, 4, or 12 animals reduced empirical accuracies for animals on test to 0.50, 0.41, and 0.21 when pooling was at random and 0.53, 0.47, and 0.34 when pooling was by sire. Simulating a fixed pen effect representing 10% phenotypic variation, but ignoring that effect in the evaluation minimally reduced empirical accuracies to 0.52, 0.46, and 0.33 when pooling by sire. Theoretical accuracies were in close agreement with empirical accuracies when the exact method was used. The approximate method that treated averages of pooled data as if they were individually observed overstated accuracy and should not be used. Selection on the basis of pooled observations can be almost as effective as using individual observations when pool sizes are small. The exact method to account for pooled data is no more complex than conventional procedures. PMID- 16361495 TI - Low birth weight is associated with enlarged muscle fiber area and impaired meat tenderness of the longissimus muscle in pigs. AB - The objective of this study was to determine the relationships between birth weight-associated modifications in histological or chemical muscle characteristics and meat quality traits in pigs. At 68 d of age, Pietrain x (Large White x Landrace) female littermates were allocated into 2 groups on the basis of low birth weight (LW = 1.05 +/- 0.04 kg; n = 15) or high birth weight (HW = 1.89 +/- 0.02 kg; n = 15). Pigs were reared in individual pens with free access to a standard diet up to slaughter at approximately 112 kg of BW. During the growing-finishing period, LW and HW pigs had a similar daily feed consumption, whereas G:F was lower (P = 0.009) for LW pigs than for HW littermates. At final BW, LW pigs were 12 d older (P < 0.001) than HW littermates. Estimated lean meat content, relative proportions of loin and ham in the carcass, and weights of LM and semitendinosus muscle (SM) were decreased (P < 0.05) in LW pigs compared with HW pigs. Conversely, the LW pigs exhibited a fatter carcass, greater activity levels of fatty acid synthase and malic enzyme in backfat (n = 15 per group), and enlarged subcutaneous adipocytes (n = 8 per group) compared with the HW pigs. Similarly, lipid content was increased by 25% (P = 0.009), and mean adipocyte diameter was 12% greater (P = 0.008) in the SM from LW pigs compared with that from HW pigs, whereas lipid content did not vary in the LM of either group. Mean myofiber cross-sectional areas were 14% greater in the LM (P = 0.045) and the SM (P = 0.062) of LW pigs than of HW pigs. Conversely, the total number of myofibers was less (P = 0.003) in the SM of LW vs. HW pigs. There were no differences between groups for glycolytic potential at slaughter and rate and extent of postmortem pH decline in both muscles, as well as for LM drip losses. A trained sensory test panel judged the roast loin meat to be less tender (P = 0.002) in LW pigs relative to HW pigs. Scores for juiciness, flavor, flouriness, and fibrousness of meat did not differ between groups. Overall, negative but somewhat low correlation coefficients were found between LM tenderness score and ultimate pH (r = -0.36; P = 0.06) and between LM tenderness and mean cross-sectional area of myofibers (r = -0.34; P = 0.07). This study demonstrates a lower tenderness of meat from pigs that had a LW, partly as a result of their enlarged myofibers at market weight. PMID- 16361496 TI - Efficacy of DL-methionine hydroxy analog free acid and DL-methionine as methionine sources for pigs. AB - Two experiments were conducted to evaluate the efficacy of dietary DL-methionine hydroxy analog-free acid (MHA-FA, 88%) compared with DL-methionine (DLM, 99%) as Met sources in pigs. In Exp. 1, a total of 245 crossbred pigs (initial BW of 6.4 kg [SD = 0.5]) were allotted to 7 treatments in 7 replicates for an experimental period of 28 d. The basal diet (BD) was formulated to contain 17.5% CP and 0.21% Met. Dietary treatments included 1) BD, 2) BD + 0.030% DLM, 3) BD + 0.060% DLM, 4) BD + 0.090% DLM, 5) BD + 0.034% MHA-FA, 6) BD + 0.068% MHA-FA, and 7) BD + 0.103% MHA-FA; the MHA-FA was supplemented on an equimolar basis to the DLM. Because of a nonlinear response, exponential regression analysis was used to evaluate the responses, and a comparison of the equations was then made to determine the relative effectiveness of the 2 Met sources. With increases in dietary Met, weight gain increased (P < 0.05). Compared with DLM on a product-to product (wt/wt) basis, the relative effectiveness of MHA-FA was calculated to be 73% for increasing weight gain and 54% for decreasing the feed:gain. In Exp. 2, a total of 30 weanling barrows [initial BW of 16.8 kg (SD = 2.8)] were used in a metabolism study to evaluate the relative value of MHA-FA to DLM. The BD was formulated to contain 16.9% CP and 0.21% Met. Dietary treatments included 1) BD, 2) BD + 0.030% DLM, 3) BD + 0.060% DLM, 4) BD + 0.046% MHA-FA, and 5) BD + 0.092% MHA-FA; the MHA-FA levels were chosen based on a pre-experiment estimate of bioequivalence in an attempt to provide approximately equal pig responses. There was no difference in fecal N output among the treatments; however, urine N linearly decreased with increasing concentrations of both sources (P = 0.034 for DLM, and P = 0.007 for MHA-FA), which resulted in a linear increase in retained N for both DLM (P = 0.012) and MHA-FA (P = 0.005). In addition, N retention (% of intake) linearly increased with increasing level of DLM (P = 0.014) and MHA-FA (P = 0.007). Using a slope-ratio procedure for comparison of the responses from the 2 sources, the relative biological equivalence value of MHA-FA to DLM in this experiment was 64.2% based on percent N retention and 66.3% based on the grams of N retained per day. Based on the results from both experiments, these data indicated that the mean relative bioequivalence of MHA-FA to DLM was 64% on a product-to-product (wt/wt) basis or 73% on an equimolar basis. PMID- 16361497 TI - Developmental changes in morphometry of the small intestine and jejunal sucrase activity during the first nine weeks of postnatal growth in pigs. AB - The objective of this study was to investigate the development of small intestinal size and digestive capacity of the jejunum in growing pigs. The weight, length, surface area, and mucosa weight of the small intestine were measured when pigs were 1, 3, 5, and 9 wk of age. Sucrase and alkaline phosphatase (ALP) activities of the jejunal brush-border membrane, prepared by differential centrifugation and Mg2+ precipitation, were determined at the respective postnatal stages. Body weights increased 7-fold from 2.7 kg at 1 wk to 23.32 kg at 9 wk postnatal. Body weight gains were greater (P < 0.05) from wk 3 to 5 than from wk 1 to 3. Weights of the small intestine and of the intestinal mucosa increased faster (P < 0.05) from 3 to 5 wk than from 1 to 3 wk; the slowest increase occurred from 5 to 9 wk. Weights of the duodenum, jejunum, and ileum, and mucosa from the respective sections increased (P < 0.05) as pigs grew from 3 to 9 wk. Mucosa weight relative to the weight of the section was greater (P < 0.05) for the duodenum and jejunum than for the ileum at 9 wk of age. Between the ages of 3 and 9 wk, the increase in mucosa weight was highest for the jejunum followed by the duodenum and the ileum. The increase was greatest for the duodenum followed by the jejunum and the ileum when mucosal weight was expressed per unit of appropriate intestinal section weight. There was a 55-fold increase in jejunal sucrase activity from 1 to 9 wk; the greatest rate of increase occurred between 5 and 9 wk. Total jejunal ALP activities in pigs at 9 wk was greater (P < 0.05) than at 5 wk, which in turn was greater than at 1 wk of age. In summary, increases in BW during the first 9 wk of postnatal growth in pigs are accompanied by significant developmental changes in digestive capacity including intestinal weights, length, and area as well as jejunal brush-border sucrase and ALP activities. PMID- 16361498 TI - Mucosal surface area and fermentation activity in the hind gut of hydrated and chronically dehydrated working donkeys. AB - The effects of mild chronic dehydration on fermentation rate and mucosal surface area in the cecum, dorsa and ventral colon, and descending colon of the hindgut were investigated in South African donkeys (n = 11) in agricultural work. Dehydration representing a 6% drop in BW (n = 6) was associated with increased fermentation activity in the cecum (252 +/- 22.9 vs. 161 +/- 13.5 micromol/g of DM.h(-1), P < 0.01) and enhanced fluid retention in the ventral colon (0.81 +/- 0.026 vs. 0.73 +/- 0.034 mL/g gut, P < 0.05). Fermentation activity in the next segment of the hindgut, the ventral colon, of dehydrated donkeys was also greater numerically (92.5 +/- 22.60 vs. 77.9 +/- 10.40 micromol/g of DM.h(-1)), but this difference was not significant. Total mucosal and crypt surface area per unit volume of gut (Sv, microm2/microm3) was greater in dehydrated donkeys for the cecum (253 +/- 23.0 vs. 161 +/- 13.5, P < 0.01), the ventral colon (286 +/- 6.2 vs. 171 +/- 9.8, P < 0.01), the dorsal colon (276 +/- 18.2 vs. 256 +/- 11.0, P < 0.05), and the descending colon (260 +/- 20.3 vs. 191 +/- 15.2, P < 0.05). Enhanced fermentation activity and enhanced mucosal absorptive or secretory capacity within the hindgut during chronic dehydration was associated with an observed maintenance of appetite. These adaptations in the hindgut are valuable physiological attributes for working donkeys in semi-arid regions where they are frequently exposed to chronic dehydration. PMID- 16361499 TI - Performance responses and indicators of gastrointestinal health in early-weaned pigs fed low-protein amino acid-supplemented diets. AB - The effects of low-protein AA-supplemented diets on piglet performance, visceral organ mass, incidence of diarrhea, intestinal microbial population, and fermentation were studied in a 3-wk trial. After a 7-d adaptation period, 96 piglets (approximately 6.2 kg of initial BW) were assigned to 4 corn-wheat, soybean meal-based dietary treatments in a completely randomized design to give 6 replicate pens per treatment (n = 4 piglets per pen). The treatments were a control wheat-corn-soybean meal-based phase I diet containing 23% CP, or the same diet with CP reduced to 21%, 19%, or 17% and supplemented with crystalline AA to achieve equal standardized ileal digestible contents of Lys, Met plus Cys, Thr, and Trp in all diets. Diets were formulated to similar nutrient levels and provided ad libitum. Blood from all pigs was taken on d 0, 7, 14, and 21 for determining plasma urea N. Weekly feed intake, BW changes, and G:F were determined. On d 21, 2 pigs per pen were randomly selected and killed to determine small intestinal morphology, digesta pH and ammonia levels, and luminal microbial counts. Average daily feed intake, ADG, and G:F were not affected (P > 0.10) by reducing CP to 21%, but a reduction to 19% or 17% decreased ADFI (P < 0.001) and ADG (linear, P < 0.001; quadratic, P < 0.05) over the 3-wk study period. Reducing CP to 19% had no effect (P > 0.10) on G:F; however, this response criterion was decreased linearly (P < 0.001) over the 3-wk study period as dietary CP declined. Water usage was only numerically decreased (P > 0.10) with dietary CP reduction. Plasma urea N was decreased linearly (P < 0.01) with CP reduction. Reducing CP from 23 to 17% had a linear (P < 0.05) and cubic effect on stomach and liver weights, respectively. Although histological data showed some differences among diets, no distinct trend was evident. Ammonia N in ileal digesta was reduced linearly (P < 0.01) as dietary CP was decreased. With the exception of valeric acid, VFA levels in ileal digesta of piglets fed low-protein diets were generally lower (P < 0.05) compared with the control diet. Diet had no effect on intestinal microbial counts (P > 0.10). The results show that piglet performance may suffer when dietary CP is reduced by 4 or more percentage units from 23% and support the hypothesis that low-CP diets help maintain enteric health in pigs by lowering toxic microbial metabolites such as ammonia. PMID- 16361500 TI - In situ studies on the time-dependent degradation of recombinant corn DNA and protein in the bovine rumen. AB - An in situ technique was adopted to investigate the time-dependent ruminal degradation of chloroplast compared with recombinant DNA of Bt176 corn using conventional and quantitative PCR assays. In parallel, the Cry1Ab protein content and fragment sizes were determined by ELISA and immunoblotting techniques. Triplicate nylon bags filled with 5 g of each substrate (whole-plant isogenic, whole-plant transgenic, ensiled isogenic, and ensiled transgenic corn) were positioned within the rumen of 5 rumen-cannulated, nonlactating cows and incubated for 2, 4, 8, 16, 24, and 48 h. To investigate the DNA degradation process, PCR assays were developed to detect fragments of the endogenous highly abundant rubisco gene (173, 896, 1,197, and 1,753 bp) and of the recombinant cry1Ab gene (211, 420, 727, and 1,423 bp). Short fragments of rubisco (<431 bp) and cry1Ab DNA (211 bp) were amplifiable in whole-plant and ensiled corn samples incubated in the rumen for 48 h, whereas the traceability of larger fragments depended on previous processing of the sample (whole-plant or ensiled corn), the length of the target sequence, and concomitantly on the length of time incubated in the rumen. Quantification of rubisco and cry1Ab gene fragments applying real time PCR assays revealed degradation to <20% of initial 0-h values within 2 h and <0.5% after 48 h of ruminal incubation. Analysis of Cry1Ab protein in whole-plant corn using the ELISA technique revealed a decrease to 28.0% of the initial value within 2 h and to 2.6% within 48 h. The concentration of Cry1Ab protein of ensiled corn was only 10% that of whole-plant corn. Ensiled corn Cry1Ab protein decreased to 10% of initial values after 48 h of ruminal incubation. Using an immunoblotting technique, the full-size Cry1Ab protein was only detectable up to 8 h; thereafter, only fragments of approximately 17 and 34 kDa size were found. In conclusion, ruminal digestion decreased the presence of functional cry1Ab gene fragments. It is unlikely that full-size, functional Cry1Ab protein will be present after 8 h of incubation in the rumen. Therefore, results based on ELISA measurements should be interpreted carefully and verified by another detection method that discriminates between the full-size and fragmented Cry1Ab protein. PMID- 16361501 TI - Relationships of feedlot feed efficiency, performance, and feeding behavior with metabolic rate, methane production, and energy partitioning in beef cattle. AB - Residual feed intake (RFI) is the difference between the actual and expected feed intake of an animal based on its BW and growth rate over a specified period. The biological mechanisms underlying the variation in feed efficiency in animals with similar BW and growth rate are not well understood. This study determined the relationship of feedlot feed efficiency, performance, and feeding behavior with digestion and energy partitioning of 27 steers. The steers were selected from a total of 306 animals based on their RFI following feedlot tests at the University of Alberta Kinsella Research Station. Selected steers were ranked into high RFI (RFI > 0.5 SD above the mean, n = 11), medium RFI (RFI +/- 0.5 SD above and below the mean, n = 8), and low RFI (RFI < -0.5 SD below the mean, n = 8). The respective BW +/- SD for the RFI groups were 495.6 +/- 12.7, 529.1 +/- 18.6, and 501.2 +/- 15.5 kg. Digestibility and calorimetry trials were performed on a corn or barley-based concentrate diet in yr 1 and 2, respectively, at 2.5 x maintenance requirements. Mean DMI (g/kg of BW(0.75)) during the measurements for high-, medium-, and low-RFI groups, respectively, were 82.7 +/- 2.0, 78.8 +/- 2.6, and 81.8 +/- 2.5 and did not differ (P > 0.10). Residual feed intake was correlated with daily methane production and energy lost as methane (r = 0.44; P < 0.05). Methane production was 28 and 24% less in low-RFI animals compared with high- and medium-RFI animals, respectively. Residual feed intake tended to be associated (P < 0.10) with apparent digestibilities of DM (r = -0.33) and CP (r = -0.34). The RFI of steers was correlated with DE (r = -0.41; P < 0.05), ME (r = 0.44; P < 0.05), heat production (HP; r = 0.68; P < 0.001), and retained energy (RE; r = -0.67; P < 0.001; energy values are expressed in kcal/kg of BW(0.75)). Feedlot partial efficiency of growth was correlated (P < 0.01) with methane production (r = -0.55), DE (r = 0.46), ME (r = 0.49), HP (r = -0.50), and RE (r = 0.62). With the exception of HP (r = 0.37; P < 0.05), feed conversion ratio was unrelated to the traits considered in the study. Feeding duration was correlated (P < 0.01) with apparent digestibility of DM (r = -0.55), CP (r = -0.47), methane production (r = 0.51), DE (r = -0.52), ME (r = -0.55), and RE (r = -0.60). These results have practical implications for the selection of animals that eat less at a similar BW and growth rate and for the environmental sustainability of beef production. PMID- 16361502 TI - Factors influencing characteristics of steam-flaked corn and utilization by finishing cattle. AB - Three experiments were conducted to identify factors influencing steam-flaked corn (SFC) characteristics and feeding value. In Exp. 1, corn samples (n = 108) were tempered for 2 h using 6, 10, or 14% moisture containing 0 or 0.67 mL of surfactant/L. Samples were steamed for 20 or 40 min and flaked to 360, 335, or 310 g/L. Treatments were arranged in a 3 x 2 x 2 x 3 factorial. No interactions existed in Exp. 1. Increasing tempering moisture linearly (P < 0.001) increased corn moisture after tempering, steaming, and flaking; however, SFC moisture was not increased (quadratic; P < 0.001) greatly by applying more than 10% water during tempering. The surfactant, steam time, and flake density had no effect (P = 0.16 to 0.93) on corn moisture after tempering, steaming, or flaking, but adding a surfactant during tempering decreased (P = 0.05) moisture loss after flaking. Starch availability was unaffected (P = 0.31 to 0.84) by tempering moisture concentration, tempering with a surfactant, or steam time but was increased (linear; P < 0.01) by decreasing flake density. Flake durability was increased by increasing tempering moisture (linear; P < 0.001), tempering with a surfactant (P = 0.04), increasing steam time (P < 0.001), and decreasing flake density (linear; P = 0.02). In Exp. 2, 89 heifers (initial BW = 350 kg) were fed 75% SFC-based diets for 108 d to determine the effects of SFC particle size on performance and carcass traits. Treatments were SFC that was mixed for 0 (4,667 microm) or 15 min (3,330 microm) before addition of other ingredients. Heifers fed 3,330-microm SFC tended (P = 0.13) to eat less DM, but ADG and G:F did not differ (P = 0.58 to 0.65) between treatments. Carcass traits did not differ, except that heifers fed 3,330-microm SFC had less (P = 0.008) KPH. In Exp. 3, 96 heifers (initial BW = 389 kg) were fed for 82 d diets containing 73% SFC that was either 18 or 36% moisture. Heifers fed 36% moisture SFC ate less DM (P = 0.02) and gained slower (P = 0.05) than heifers fed 18% moisture SFC, but G:F did not differ (P = 0.93) with SFC moisture. Heifers fed 36% moisture SFC were fatter at the 12th rib (P = 0.009), but all other carcass traits did not differ. Methods that increase moisture of SFC improved durability, but extreme moisture levels negatively affected cattle performance. Flake particle size did not affect cattle performance. Flake density is the major factor affecting starch availability in SFC. PMID- 16361503 TI - Effect of refined coconut oil or copra meal on methane output and on intake and performance of beef heifers. AB - An experiment was conducted to establish the effect of feeding either refined coconut oil (CO) or copra meal containing CO to beef heifers on DMI, animal performance, enteric CH4 emissions, diet digestibility, and the fatty acid profile of the resulting meat. Forty-one Charolais and Limousin crossbred beef heifers (474 +/- 29 kg; 661 +/- 89 d of age) were blocked by BW before being assigned in a randomized complete block design to 1 of 3 experimental treatments (n = 12) or to a pretrial slaughter group (n = 5) used to determine the initial carcass weight. The experimental period lasted for 93 d. Enteric CH4 output was recorded for 2 periods of 5 consecutive days from d 14 to 18 and from d 70 to 74. The 3 dietary treatments were 1) control, a barley/soybean meal-based concentrate with 0 g of CO/ d; 2) RCO, a barley/soybean meal-based concentrate with 250 g of CO/d from refined coconut oil; and 3) CM, a copra meal-based concentrate with 250 g of CO/d from copra meal. Each diet had a 50:50 forage:concentrate using grass silage as the forage source. There was no effect of diet on DMI (P = 0.734) or GE intake (P = 0.486). The addition of RCO increased ADG (P < 0.05) compared with the control treatment. The CM treatment decreased (P < 0.05) average daily carcass gain compared with the RCO treatment only. There was a decrease (P < 0.05) in the digestibility of the DM, OM, CP, and GE fractions of the diet only with the CM treatment. Both the RCO and CM concentrates decreased (P < 0.001) daily enteric CH4 output when expressed in terms of liters per day, liters per kilogram of DMI, percentage of GE intake, liters per kilogram of ADG, and liters per kilogram of average daily carcass gain. The RCO treatment produced the greatest numerical response for all measures. Ruminal protozoa numbers on the RCO treatment were lower (P < 0.05) than on the control treatment. The concentrations of the fatty acid methyl esters, lauric (P < 0.001) and myristic (P < 0.002) acids, were increased in muscle when either of the CCO treatments was compared with the controls, but the differences were of a magnitude unlikely to influence human health status. Although the CM concentrate decreased CH4 comparable with the RCO concentrate, decreased performance resulted in an extended finishing time with implications for lifetime CH4 emissions. PMID- 16361504 TI - The effects of high levels of supplemental copper on the serum lipid profile, carcass traits, and carcass composition of goat kids. AB - This experiment was conducted to determine the effect of high levels of supplemental Cu (as (CuSO4).5H2O) on the serum lipid profile and carcass traits of goat kids. Fifteen Boer x Spanish wether goat kids (BW = 21.3 +/- 0.7 kg) were housed in individual pens and were assigned randomly to 1 of 3 treatments. Treatments consisted of 1) control (no additional supplemental Cu), 2) 100 mg of Cu/d, and 3) 200 mg of Cu/d. Copper sulfate was placed in gelatin capsules and inserted into the esophagus via a balling gun before the morning feeding. Animals were fed a high-concentrate (70:30 grain:hay) diet for 112 d. Serum lipid profile was determined on d 14 and 112, and BW was recorded after 4-h withdrawals from feed and water. After 112 d, animals were slaughtered, and carcass traits were measured. The left half of 12 carcasses and 9th to 11th rib sections from the right side of 15 carcasses were dissected into separable soft tissue and bone portions. The soft tissue portion was analyzed for moisture, ether extract, CP, and ash. Average daily feed intake decreased (linear; P = 0.05), and G:F increased (quadratic; P = 0.02) in the 100 mg of Cu/d group. Serum cholesterol and triglycerides did not change (P > 0.10); however, NEFA decreased (linear; P = 0.01) as supplemental Cu increased. No differences were observed (P > 0.10) in HCW, chilled carcass weight, or kidney and pelvic fat; however, 12th rib fat (linear; P = 0.01) and adjusted fat thickness (linear; P = 0.03) decreased as Cu supplementation increased. No differences (P > 0.10) in LM area were observed; however, percentage of boneless closely trimmed retail cuts increased (linear; P = 0.04) as Cu supplementation increased. The moisture (%) of the 9th to 11th rib sections increased (linear; P = 0.03), ether extract (%) decreased (linear; P = 0.02), and CP and ash (%) tended to increase (linear; P = 0.09 and 0.06, respectively) as Cu supplementation increased. Carcass composition measured using the left half of the carcass confirmed the values obtained through the 9th to 11th rib sections. Results of this study indicate that supplemental Cu can alter the serum lipid profile, carcass characteristics, and carcass composition of goat kids. PMID- 16361505 TI - Influence of harvest processes on pork loin and ham quality. AB - The purpose of this study was to determine the specific effects of extending the interval between dwell time and the duration of scalding on pork quality attributes. Sixty-four Duroc x Yorkshire pigs were randomly assigned to a 2 x 2 factorial treatment arrangement. Treatments included extending the dwell duration from 5 to 10 min and extending the scald duration from 5 to 8 min. All carcasses entered the cooler 50 min after exsanguination. At exsanguination, blood was collected for three 1-min intervals and then for a final 2-min period. Temperature and pH of the LM and semimembranosus muscle (SM) were measured at 45 min, and at 2, 4, 6, and 24 h postmortem (PM). Hunter L*, a*, and b* values were determined on the LM, SM, and biceps femoris (BF). Purge loss was measured on the SM, BF, and the sirloin end of the loin. Drip loss was measured in duplicate from LM chops after 1 and 5 d of storage. Warner-Bratzler shear force (WBS) measurements were determined on LM chops aged 1, 3, 5, and 7 d PM. Over 99% of the collected blood was obtained during the first 3 min after sticking. Carcasses scalded for 8 min had greater (P < 0.05) semi-membranosus 2 h temperature (28.8 degrees C) than carcasses scalded 5 min (27.3 degrees C). An 8-min scald process resulted in longissimus dorsi chops with lower hue angle and greater WBS values than the 5-min scald process. Increasing dwell time from 5 to 10 min resulted in biceps femoris chops with greater hue angle and loin chops with greater WBS values at 3 d PM. Harvest processes did not significantly affect subjective quality scores, Hunter L* values, purge or drip loss. Lengthening the duration of dwell and scalding may result in a more rapid PM pH decline. Reducing the duration of scalding may lead to increased time for manual removal of hair. Because of differences in facilities, it is recommended that individual facilities monitor dwell and scald durations to determine how to best minimize time of entry into the cooler. PMID- 16361506 TI - Effects of supplemental magnesium concentration of drinking water on pork quality. AB - Thirty-two barrows were used to determine the effects of supplemental Mg in drinking water on pork quality. Pigs were determined to be free of the halothane and Napole mutations and were individually penned. After a 7-d adjustment period, barrows (111 +/- 1 kg BW) were blocked by BW and allotted randomly within block to 0, 300, 600, or 900 mg of supplemental Mg from Mg sulfate/L of drinking water for 2 d before slaughter. Pigs were not allowed access to feed (0.13% Mg) for 15 h before slaughter but continued to have access to experimental water treatments. Pigs were loaded and transported 110 km (1.75 h) to a commercial abattoir and remained in lairage for 5 h before slaughter. The LM was removed 24 h postmortem. Retail storage was simulated for 8 d, and the remaining LM was vacuum-packaged for 25 or 50 d at 4 degrees C. Plasma Mg concentration increased linearly (P = 0.001) with Mg supplementation; however, Mg concentration of the LM was not affected (P = 0.99) by Mg supplementation. Surface exudate, drip loss, and retail fluid loss of the LM were not affected (P > 0.10) by Mg. Lightness (L*) and redness (a*) of the LM were not affected (P > 0.10) by Mg, with the exception of initial redness (cubic; P = 0.05). Pigs supplemented with 300 or 900 mg of Mg/L had lower yellowness (b*) values of the LM displayed for 0 to 6 d than pigs supplemented with 0 or 600 mg of Mg/L (cubic; P < 0.05). Lightness of the LM after 25 (quadratic; P = 0.03) or 50 (quadratic; P = 0.04) d of vacuum-packed storage was greater at 300 and 600 mg of Mg/L than at 0 or 900 mg/L. Yellowness tended to be greater after 50 d, but not after 25 d, of vacuum-packaged storage for 300 or 600 mg of Mg/L compared with 0 or 900 mg/L (quadratic; P = 0.08). Oxidation of the LM, determined by thiobarbituric acid reactive substances after 4 d of retail storage, increased linearly (P = 0.05) as Mg increased in the drinking water. Furthermore, oxidation of the LM after 8 d of retail storage tended to increase linearly (P < 0.10), primarily because of the high oxidation of LM from pigs supplemented with 900 mg of Mg/L compared with controls (224 vs. 171 +/- 19 microg/kg, respectively). Oxidation of the LM was greater for pigs supplemented with 300 or 900 mg/L compared with 0 or 600 mg of Mg/L (cubic; P < 0.06) after 25 d of vacuum-packed storage. Magnesium did not improve pork quality characteristics of practical significance in pigs without the halothane and Rendement Napole mutations. PMID- 16361507 TI - Effects of available dietary carbohydrate and preslaughter treatment on glycolytic potential, protein degradation, and quality traits of pig muscles. AB - The current study was conducted to determine the interactive effects of a glycogen-reducing diet fed to finishing pigs and length of preslaughter transportion on muscle metabolic traits, proteolysis of intermediate filament and costameric proteins, and meat quality traits. Large White gilts and barrows (n = 48) were selected at 88 kg of BW and individually fed for 21 d a diet (2.6 kg/d) either high (HC) or low (LC) in available carbohydrates. Six gilts and 6 barrows fed the HC and LC diets were subjected to 0 or 3 h of transportation on the day of slaughter. Muscle temperature and pH were measured at 0.5, 1.5, 2.5, 3.5, 4.5, 5.5, and 24 h postmortem in the LM and 24 h postmortem in the dark (STD) and light (STL) portion of the semitendinosus. At 24 h postmortem, glycolytic potential (GP) was determined in the LM, STD, and STL, as well as proteolysis of titin, nebulin, desmin, vinculin, and talin in the LM and STD. The GP was lower (P < 0.05) in muscles from LC-pigs than in muscles from HC-pigs. The LC diet also resulted in lower (P < 0.05) pH, and a darker (P = 0.03), less (P < 0.01) yellow color in the STL. The LC diet decreased (P = 0.04) cooking losses in the STL and STD. The 3-h journey further decreased (P = 0.05) the GP in the STD, regardless of the diet, but transport had no effect (P > or = 0.67) on the GP of the LM and STL. Ultimate pH of the LM was lower (P = 0.02), and both portions of the semitendinosus were darker (P = 0.01) and less yellow (P < 0.01), in pigs transported 3 vs. 0 h. In pigs transported for 3 h, intact vinculin tended to be more (P = 0.08) degraded in the LM, which coincided with lower (P = 0.04) drip losses in the LM of pigs transported for 3 compared with 0 h. Increased (P < 0.01) proteolysis of titin paralleled lower (P = 0.02) shear force values in the STD of pigs transported 3 vs. 0 h. Although the present results demonstrated the potential of a glycogen-reducing diet to alter the GP of different porcine muscles, the effect of these changes on meat quality traits was limited to higher ultimate pH and darker color in the STL. The positive effects of length of transportation on water-holding capacity (LM and STD) and meat color (STD and STL) were only partially related to the resting muscle glycogen concentration because the 3-h transport lowered the GP only in the muscle with the lowest basal glycogen concentration. PMID- 16361508 TI - Cortisol-binding globulin and meat quality in five European lines of pigs. AB - The gene (Cbg) encoding cortisol-binding globulin (CBG) has been proposed as a candidate gene to explain genetic variation in cortisol secretion and carcass composition in pigs. The objective of this study was to evaluate the association between CBG and pork quality in 5 European breeding lines, Pietrain, Large White (LW), and Landrace purebred lines, a Duroc synthetic line, and a Meishan (MS) x LW advanced intercross. Cortisol-binding globulin maximum binding capacity (CBG Bmax) was twice as high (P < 0.05) in MS x LW pigs compared with the other lines. There was no (P > or = 0.364) association between CBG-Bmax and carcass quality traits in Pietrain gilts, but CBG-Bmax was associated with increased loin yields in LW (P = 0.010) and Landrace (P = 0.103) gilts, decreased ham yields (P = 0.082) in Duroc gilts, and increased fat depth (P = 0.064) and leaf fat (P = 0.001) in MS x LW gilts. There was no association between CBG-Bmax and pork quality traits in Pietrain (P > or = 0.269) and Duroc (P > or = 0.114) gilts. Conversely, CBG-Bmax was associated with lighter (higher L* values; P < 0.05) pork in Land-race gilts, as well as lower (P < or = 0.055) ultimate pH in the LM and semimembranosus, and a tendency for lower (P = 0.095) L* values of pork from LW gilts. Within MS x LW pigs, CBG-Bmax was associated with increased drip loss (P = 0.001) and decreased i.m. fat in the semimembranosus (P = 0.005). Because drip loss is an economically important pork quality trait, results of this study could be used in the selection of improved water-holding capacity of pork from synthetic lines involving the MS breed. PMID- 16361509 TI - Use of supplementary tryptophan to modify the behavior of pigs. AB - Three experiments were conducted to investigate the short-term use of supplementary Trp on the behavior of grow/finish pigs. Three levels of dietary Trp were used, representing the standard requirement for growth (control), twice (2x), and 4 times (4x) the control amount. In Exp. 1, pigs were fed the diets for 7 d, during which observations were made of their general behavior (time budget), aggression within the group of familiar pigs, and response to a startling auditory stimulus. Behavior effects were evident during the period of supplementation for both the 2x and 4x diets. During the treatment period, pigs fed supplemental Trp spent more time lying (P = 0.04) and less time eating (P = 0.05) than pigs fed the control diet. Although the response of the animals to the startling stimulus was to become alert and stand, similar behavioral effects caused by supplemental Trp also were evident after the startling stimulus (P < 0.01). Based on these observations, the subsequent studies retained the same dietary levels of Trp and incorporated a 3-d feeding of diets before behavior testing. In Exp. 2, pigs were fed the experimental diets for 3 d before being regrouped with unfamiliar pigs on the same diet. Subsequent aggression was affected by Trp supplementation, in that high levels of dietary Trp decreased the total duration of fighting by approximately 50% (P = 0.03). Supplemental Trp had no effect on the number of fights, and there were no differences between the 2 levels of supplemental Trp on any behavior. In Exp. 3, pigs were exposed to specific handling tests on the farm and meat quality assessments after being fed the experimental diets for 3 d. There were no differences among dietary treatments for any of the meat quality characteristic variables measured. The only behavioral or physiological difference observed among the treatments was a slower movement of pigs fed the 4x Trp treatment than control or 2x Trp-fed pigs in a minimal-forced situation (P = 0.04). Response to confinement on a scale, an electric prod, and movement in general did not differ among treatments. High levels of Trp may result in animals avoiding stressful situations if possible, but they seem to have no effect on responses to stressors that animals may experience in a forced situation. PMID- 16361510 TI - Evaluation of pet food by-product as an alternative feedstuff in weanling pig diets. AB - Three experiments were conducted to evaluate pet food by-product (PFB) as a component of nursery starter diets and its effects on pig performance. The PFB used in these studies was a pelleted dog food that contained (as-fed basis) 21% CP, 1.25% total lysine, and 8.3% ether extract. In Exp. 1, 288 early-weaned pigs (5.2 kg at 14 d) were used to determine the effects of replacing animal protein and energy sources with PFB at 0, 10, 30, and 50% (as-fed basis) inclusion levels in phase I (d 0 to 7 after weaning) and phase II (d 7 to 21 after weaning) diets. Phase I diets contained 27.5% whey, 18.75% soybean meal, 1.50% lysine, 0.90% Ca, and 0.80% P, with PFB substituted for corn, fat, plasma protein, fish meal, limestone, and dicalcium phosphate. Phase II diets had a constant 10% whey, 1.35% lysine, and PFB was substituted for blood cells, a portion of the soybean meal, and other ingredients as in phase I diets. In phase I, growth performance by pigs fed PFB-containing diets was similar to that of the control diet. In phase II, ADG (linear; P < 0.05 and quadratic, P < 0.005), ADFI (linear and quadratic, P < 0.01), and G:F (quadratic, P < 0.01) were increased with increasing PFB inclusion. In Exp. 2, 80 weaned pigs (6.7 kg at 21 d) were fed a common phase I diet for 1 wk and used to further evaluate the effect of PFB in phase II diets (same as Exp 1; initial BW = 8.1 kg) on growth performance and apparent total tract nutrient digestibility. There were no differences in ADG, ADFI, or G:F across treatments. Dry matter and energy digestibility did not differ among diets; however, digestibilities of CP (P < 0.05) and the essential AA, arginine (P < 0.02), histidine (P < 0.01), lysine (P < 0.001), threonine (P < 0.01), and valine (P < 0.01), were greater as PFB was increased in the diet. In Exp. 3, the performance by pigs (n = 1 70; 5.5 kg; 21 d of age) fed diets with 0 or 30% PFB in both phases I and II was examined. Growth performance was similar in both diets. These studies demonstrate that pet food by-product can effectively be used as a partial replacement for animal protein sources and grain energy sources in the diets of young nursery pigs. PMID- 16361511 TI - Application of broken-line analysis to assess floor space requirements of nursery and grower-finisher pigs expressed on an allometric basis. AB - Few issues in swine production are as complex as floor space allowances. One method for pork producers to calculate floor space allowance (A) is to convert BW into a 2-dimensional concept yielding an expression of A = k * BW(0.667). Data on ADG, ADFI, and G:F were obtained from published peer-reviewed studies. Five data sets were created: A = grower-finisher pigs, fully slatted floors, and consistent group size; B = grower-finisher pigs and fully slatted floors (group size did not need to be consistent); C = grower-finisher pigs, partially slatted floors, and consistent group size; D = grower-finisher pigs, partially slatted floors (group size did not need to be consistent); and E = nursery pigs, fully slatted or woven wire floors (group size did not need to be consistent). Each data set was analyzed using a broken-line analysis and a linear regression. For the broken line analyses, the critical k value, below which a decrease in ADG occurred, varied from 0.0317 to 0.0348. In all cases the effect of space allowance on ADG was significant (P < 0.05). Using the linear analyses based on data with k values of < 0.030, the critical k values for the 4 grower-finisher data sets did not differ from those obtained using the broken-line analysis (0.0358 vs. 0.0336, respectively; P > 0.10); however, none of the linear regressions explained a significant proportion of the variation in ADG. The slopes for the nonplateau portion of the broken-line analyses based on percent values varied among data sets. For every 0.001 decrease in k (approximately 3% of the critical k value), ADG decreased by 0.56 to 1.41%, with an average value of 0.98% for the 5%-based analyses. The use of an allometric approach to express space allowance and broken line analysis to establish space requirements seem to be useful tools for pig production. The critical k value at which crowding becomes detrimental to the growth of the pig is similar in full- and partial-slat systems and in nursery and grower-finisher stages. The critical point for crowding determined in these analyses approximated current recommendations to ensure the welfare of pigs. PMID- 16361512 TI - Effects of diets with cereal grains contaminated by graded levels of two Fusarium toxins on selected immunological and histological measurements in the spleen of gilts. AB - Feeding experiments with diets containing Fusarium toxin-contaminated wheat were conducted to clarify the pathogenesis of immunological effects of Fusarium toxins to porcine spleen cells. Contaminated diets were fed to 36 Landrace prepubertal gilts for 35 d. Concentrations (as-fed basis) of the indicator toxins deoxynivalenol (DON) and zearalenone (ZON), respectively, were 210 and 4 (control -group I), 3,070 and 88 (group II), 6,100 and 235 (group III), and 9,570 and 358 microg/kg (group IV). No signs of hyperestrogenism or uterotrophic effects were observed because of dietary treatments. The feeding of mycotoxin-contaminated diets did not cause gross pathological findings in the spleens of animals. In vivo, no inhibitory effects were detected on concanavalin A-stimulation of blood lymphocytes; however, the proliferation rate of splenocytes was inhibited (P < 0.05) in pigs fed the diet with the highest DON/ZON concentration. With in vitro studies, lower proliferation rates of blood lymphocytes and splenocytes preexposed to DON were detected. Serum IgA concentrations of pigs in group II were increased (P < 0.05) compared with the baseline value before feeding the DON/ZON diet. The histopathological data indicated elevated (P < 0.05) iron staining in the red pulp of spleens in gilts from groups I to IV after 35 d of feeding. The presence of hemosiderin particles in the spleen sections was confirmed by transmission electron microscopic investigation. Together, the results provide evidence of spleen dysfunction (hemosiderosis) in the absence of clinical signs, especially in pigs fed higher concentrations (groups III and IV) of Fusarium toxin-contaminated wheat. PMID- 16361514 TI - The Arabidopsis group 1 LATE EMBRYOGENESIS ABUNDANT protein ATEM6 is required for normal seed development. AB - As part of the embryo maturation process, orthodox seeds undergo a developmentally regulated dehydration period. The LATE EMBRYOGENESIS ABUNDANT (LEA) genes encode a large and diverse family of proteins expressed during this time. Many hypothesize that LEA proteins act by mitigating water loss and maintaining cellular stability within the desiccated seed, although the mechanisms of their actions remain largely unknown. The model plant Arabidopsis (Arabidopsis thaliana) contains two genes belonging to the group 1 LEA family, ATEM1 and ATEM6, and knockout mutations in these genes are being sought as a means to better understand group 1 LEA protein function during embryo maturation. We have identified a T-DNA insertion allele of the ATEM6 gene in which the T-DNA is present just downstream of the protein coding region. While this gene is transcriptionally active and encodes a wild-type protein, there is no detectable ATEM6 protein in mature seeds. Mutant seeds display premature seed dehydration and maturation at the distal end of siliques, demonstrating that this protein is required for normal seed development. We propose that one function for group 1 LEA proteins in seed development is to buffer the water loss that occurs during embryo maturation and that loss of ATEM6 expression results in the mutant phenotype. PMID- 16361515 TI - An LL-diaminopimelate aminotransferase defines a novel variant of the lysine biosynthesis pathway in plants. AB - Although lysine (Lys) biosynthesis in plants is known to occur by way of a pathway that utilizes diaminopimelic acid (DAP) as a central intermediate, the available evidence suggests that none of the known DAP-pathway variants found in nature occur in plants. A new Lys biosynthesis pathway has been identified in Arabidopsis (Arabidopsis thaliana) that utilizes a novel transaminase that specifically catalyzes the interconversion of tetrahydrodipicolinate and LL diaminopimelate, a reaction requiring three enzymes in the DAP-pathway variant found in Escherichia coli. The LL-DAP aminotransferase encoded by locus At4g33680 was able to complement the dapD and dapE mutants of E. coli. This result, in conjunction with the kinetic properties and substrate specificity of the enzyme, indicated that LL-DAP aminotransferase functions in the Lys biosynthetic direction under in vivo conditions. Orthologs of At4g33680 were identified in all the cyanobacterial species whose genomes have been sequenced. The Synechocystis sp. ortholog encoded by locus sll0480 showed the same functional properties as At4g33680. These results demonstrate that the Lys biosynthesis pathway in plants and cyanobacteria is distinct from the pathways that have so far been defined in microorganisms. PMID- 16361516 TI - Overexpression of RAN1 in rice and Arabidopsis alters primordial meristem, mitotic progress, and sensitivity to auxin. AB - Ran is an evolutionarily conserved eukaryotic GTPase. We previously identified a cDNA of TaRAN1, a novel Ran GTPase homologous gene in wheat (Triticum aestivum) and demonstrated that TaRAN1 is associated with regulation of genome integrity and cell division in yeast (Saccharomyces cerevisiae) systems. However, much less is known about the function of RAN in plant development. To analyze the possible biological roles of Ran GTPase, we overexpressed TaRAN1 in transgenic Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa). TaRAN1 overexpression increased the proportion of cells in the G2 phase of the cell cycle, which resulted in an elevated mitotic index and prolonged life cycle. Furthermore, it led to increased primordial tissue, reduced number of lateral roots, and stimulated hypersensitivity to exogenous auxin. The results suggest that Ran protein was involved in the regulation of mitotic progress, either in the shoot apical meristem or the root meristem zone in plants, where auxin signaling is involved. This article determines the function of RAN in plant development mediated by the cell cycle and its novel role in meristem initiation mediated by auxin signaling. PMID- 16361517 TI - Distinct roles of the first introns on the expression of Arabidopsis profilin gene family members. AB - Profilin is a small actin-binding protein that regulates cellular dynamics of the actin cytoskeleton. In Arabidopsis (Arabidopsis thaliana), five profilins were identified. The vegetative class profilins, PRF1, PRF2, and PRF3, are expressed in vegetative organs. The reproductive class profilins, PRF4 and PRF5, are mainly expressed in pollen. In this study, we examined the role of the first intron in the expression of the Arabidopsis profilin gene family using transgenic plants and a transient expression system. In transgenic plants, we examined PRF2 and PRF5, which represent vegetative and reproductive profilins. The expression of the PRF2 promoter fused with the beta-glucuronidase (GUS) gene was observed in the vascular bundles, but transgenic plants carrying the PRF2 promoter-GUS with its first intron showed constitutive expression throughout the vegetative tissues. However, the first intron of PRF5 had little effect on the reporter gene expression pattern. Transgenic plants containing PRF5 promoter-GUS fusion with or without its first intron showed reproductive tissue-specific expression. To further investigate the different roles of the first two introns on gene expression, the first introns were exchanged between PRF2 and PRF5. The first intron of PRF5 had no apparent effect on the expression pattern of the PRF2 promoter. But, unlike the intron of PRF5, the first intron of PRF2 greatly affected the reproductive tissue-specific expression of the PRF5 promoter, confirming a different role for these introns. The results of a transient expression assay indicated that the first intron of PRF1 and PRF2 enhances gene expression, whereas PRF4 and PRF5 do not. These results suggest that the first introns of profilin genes are functionally distinctive and the first introns are required for the strong and constitutive gene expression of PRF1 and PRF2 in vegetative tissues. PMID- 16361518 TI - Repressing the expression of the SUCROSE NONFERMENTING-1-RELATED PROTEIN KINASE gene in pea embryo causes pleiotropic defects of maturation similar to an abscisic acid-insensitive phenotype. AB - The classic role of SUCROSE NONFERMENTING-1 (Snf1)-like kinases in eukaryotes is to adapt metabolism to environmental conditions such as nutrition, energy, and stress. During pea (Pisum sativum) seed maturation, developmental programs of growing embryos are adjusted to changing physiological and metabolic conditions. To understand regulation of the switch from cell proliferation to differentiation, SUCROSE NONFERMENTING-1-RELATED PROTEIN KINASE (SnRK1) was antisense repressed in pea seeds. Transgenic seeds show maturation defects, reduced conversion of sucrose into storage products, lower globulin content, frequently altered cotyledon surface, shape, and symmetry, as well as occasional precocious germination. Gene expression analysis of embryos using macroarrays of 5,548 seed-specific genes revealed 183 differentially expressed genes in two clusters, either delayed down-regulated or delayed up-regulated during transition. Delayed down-regulated genes are related to mitotic activity, gibberellic acid/brassinosteroid synthesis, stress response, and Ca2+ signal transduction. This specifies a developmentally younger status and conditional stress. Higher gene expression related to respiration/gluconeogenesis/fermentation is consistent with a role of SnRK1 in repressing energy-consuming processes in maturing cotyledons under low oxygen/energy availability. Delayed up-regulated genes are mainly related to storage protein synthesis and stress tolerance. Most of the phenotype resembles abscisic acid (ABA) insensitivity and may be explained by reduced Abi-3 expression. This may cause a reduction in ABA functions and/or a disconnection between metabolic and ABA signals, suggesting that SnRK1 is a mediator of ABA functions during pea seed maturation. SnRK1 repression also impairs gene expression associated with differentiation, independent from ABA functions, like regulation and signaling of developmental events, chromatin reorganization, cell wall synthesis, biosynthetic activity of plastids, and regulated proteolysis. PMID- 16361519 TI - Cell type-specific role of the retinoblastoma/E2F pathway during Arabidopsis leaf development. AB - Organogenesis in plants is almost entirely a postembryonic process. This unique feature implies a strict coupling of cell proliferation and differentiation, including cell division, arrest, cell cycle reactivation, endoreplication, and differentiation. The plant retinoblastoma-related (RBR) protein modulates the activity of E2F transcription factors to restrict cell proliferation. Arabidopsis contains a single RBR gene, and its loss of function precludes gamete formation and early development. To determine the relevance of the RBR/E2F pathway during organogenesis, outside its involvement in cell division, we have used an inducible system to inactivate RBR function and release E2F activity. Here, we have focused on leaves where cell proliferation and differentiation are temporally and developmentally regulated. Our results reveal that RBR restricts cell division early during leaf development when cell proliferation predominates, while it regulates endocycle occurrence at later stages. Moreover, shortly after leaving the cell cycle, most of leaf epidermal pavement cells retain the ability to reenter the cell cycle and proliferate, but maintain epidermal cell fate. On the contrary, mesophyll cells in the inner layers do not respond in this way to RBR loss of activity. We conclude that there exists a distinct response of different cells to RBR inactivation in terms of maintaining the balance between cell division and endoreplication during Arabidopsis (Arabidopsis thaliana) leaf development. PMID- 16361520 TI - Role of petal-specific orcinol O-methyltransferases in the evolution of rose scent. AB - Orcinol O-methyltransferase (OOMT) 1 and 2 catalyze the last two steps of the biosynthetic pathway leading to the phenolic methyl ether 3,5-dimethoxytoluene (DMT), the major scent compound of many rose (Rosa x hybrida) varieties. Modern roses are descended from both European and Chinese species, the latter being producers of phenolic methyl ethers but not the former. Here we investigated why phenolic methyl ether production occurs in some but not all rose varieties. In DMT-producing varieties, OOMTs were shown to be localized specifically in the petal, predominantly in the adaxial epidermal cells. In these cells, OOMTs become increasingly associated with membranes during petal development, suggesting that the scent biosynthesis pathway catalyzed by these enzymes may be directly linked to the cells' secretory machinery. OOMT gene sequences were detected in two non DMT-producing rose species of European origin, but no mRNA transcripts were detected, and these varieties lacked both OOMT protein and enzyme activity. These data indicate that up-regulation of OOMT gene expression may have been a critical step in the evolution of scent production in roses. PMID- 16361521 TI - Grapes on steroids. Brassinosteroids are involved in grape berry ripening. AB - Fruit ripening is a unique plant developmental process with direct implications for our food supply, nutrition, and health. In contrast to climacteric fruit, where ethylene is pivotal, the hormonal control of ripening in nonclimacteric fruit, such as grape (Vitis vinifera), is poorly understood. Brassinosteroids (BRs) are steroidal hormones, essential for normal plant growth and development but not previously implicated in the ripening of nonclimacteric fruit. Here we show that increases in endogenous BR levels, but not indole-3-acetic acid (IAA) or GA levels, are associated with ripening in grapes. Putative grape homologs of genes encoding BR biosynthesis enzymes (BRASSINOSTEROID-6-OXIDASE and DWARF1) and the BR receptor (BRASSINOSTEROID INSENSITIVE 1) were isolated, and the function of the grape BRASSINOSTEROID-6-OXIDASE gene was confirmed by transgenic complementation of the tomato (Lycopersicon esculentum) extreme dwarf (dx/dx) mutant. Expression analysis of these genes during berry development revealed transcript accumulation patterns that were consistent with a dramatic increase in endogenous BR levels observed at the onset of fruit ripening. Furthermore, we show that application of BRs to grape berries significantly promoted ripening, while brassinazole, an inhibitor of BR biosynthesis, significantly delayed fruit ripening. These results provide evidence that changes in endogenous BR levels influence this key developmental process. This may provide a significant insight into the mechanism controlling ripening in grapes, which has direct implications for the logistics of grape production and down-stream processing. PMID- 16361522 TI - The protein phosphatase AtPP2CA negatively regulates abscisic acid signal transduction in Arabidopsis, and effects of abh1 on AtPP2CA mRNA. AB - To identify new loci in abscisic acid (ABA) signaling, we screened a library of 35ScDNA Arabidopsis (Arabidopsis thaliana)-expressing lines for ABA-insensitive mutants in seed germination assays. One of the identified mutants germinated on 2.5 microm ABA, a concentration that completely inhibits wild-type seed germination. Backcrosses and F2 analyses indicated that the mutant exhibits a dominant phenotype and that the ABA insensitivity was linked to a single T-DNA insertion containing a 35ScDNA fusion. The inserted cDNA corresponds to a full length cDNA of the AtPP2CA gene, encoding a protein phosphatase type 2C (PP2C). Northern-blot analyses demonstrated that the AtPP2CA transcript is indeed overexpressed in the mutant (named PP2CAox). Two independent homozygous T-DNA insertion lines, pp2ca-1 and pp2ca-2, were recovered from the Arabidopsis Biological Resource Center and shown to lack full-length AtPP2CA expression. A detailed characterization of PP2CAox and the T-DNA disruption mutants demonstrated that, whereas ectopic expression of a 35SAtPP2CA fusion caused ABA insensitivity in seed germination and ABA-induced stomatal closure responses, disruption mutants displayed the opposite phenotype, namely, strong ABA hypersensitivity. Thus our data demonstrate that the PP2CA protein phosphatase is a strong negative regulator of ABA signal transduction. Furthermore, it has been previously shown that the AtPP2CA transcript is down-regulated in the ABA hypersensitive nuclear mRNA cap-binding protein mutant abh1. We show here that down-regulation of AtPP2CA in abh1 is not due to impaired RNA splicing of AtPP2CA pre-mRNA. Moreover, expression of a 35SAtPP2CA cDNA fusion in abh1 partially suppresses abh1 hypersensitivity, and the data further suggest that additional mechanisms contribute to ABA hypersensitivity of abh1. PMID- 16361523 TI - The regulator of G-protein signaling proteins involved in sugar and abscisic acid signaling in Arabidopsis seed germination. AB - The regulator of G-protein signaling (RGS) proteins, recently identified in Arabidopsis (Arabidopsis thaliana; named as AtRGS1), has a predicted seven transmembrane structure as well as an RGS box with GTPase-accelerating activity and thus desensitizes the G-protein-mediated signaling. The roles of AtRGS1 proteins in Arabidopsis seed germination and their possible interactions with sugars and abscisic acid (ABA) were investigated in this study. Using seeds that carry a null mutation in the genes encoding RGS protein (AtRGS1) and the alpha subunit (AtGPA1) of the G protein in Arabidopsis (named rgs1-2 and gpa1-3, respectively), our genetic evidence proved the involvement of the AtRGS1 protein in the modulation of seed germination. In contrast to wild-type Columbia-0 and gpa1-3, stratification was found not to be required and the after-ripening process had no effect on the rgs1-2 seed germination. In addition, rgs1-2 seed germination was insensitive to glucose (Glc) and sucrose. The insensitivities of rgs1-2 to Glc and sucrose were not due to a possible osmotic stress because the germination of rgs1-2 mutant seeds showed the same response as those of gpa1-3 mutants and wild type when treated with the same concentrations of mannitol and sorbitol. The gpa1-3 seed germination was hypersensitive while rgs1-2 was less sensitive to exogenous ABA. The different responses to ABA largely diminished and the inhibitory effects on seed germination by exogenous ABA and Glc were markedly alleviated when endogenous ABA biosynthesis was inhibited. Hypersensitive responses of seed germination to both Glc and ABA were also observed in the overexpressor of AtRGS1. Analysis of the active endogenous ABA levels and the expression of NCED3 and ABA2 genes showed that Glc significantly stimulated the ABA biosynthesis and increased the expression of NCED3 and ABA2 genes in germinating Columbia seeds, but not in rgs1-2 mutant seeds. These data suggest that AtRGS1 proteins are involved in the regulation of seed germination. The hyposensitivity of rgs1-2 mutant seed germination to Glc might be the result of the impairment of ABA biosynthesis during seed germination. PMID- 16361524 TI - Increased accumulation of cuticular wax and expression of lipid transfer protein in response to periodic drying events in leaves of tree tobacco. AB - Cuticular wax deposition and composition affects drought tolerance and yield in plants. We examined the relationship between wax and dehydration stress by characterizing the leaf cuticular wax of tree tobacco (Nicotiana glauca L. Graham) grown under periodic dehydration stress. Total leaf cuticular wax load increased after each of three periods of dehydration stress using a CH2Cl2 extraction process. Overall, total wax load increased 1.5- to 2.5-fold, but composition of the wax was not altered. Homologous series of wax components were classified into organic groups; n-hentriacontane was the largest component (>75%) with alcohols and fatty acids representing <10% of the entire wax load. An increase in density, but no change in the three-dimensional shape, of leaf wax crystals was evident under low-kV scanning electron microscopy after each drying event. Leaves excised from plants subjected to multiple drying events were more resistant to water loss compared to leaves excised from well-watered plants, indicating that there is a negative relationship between total wax load and epidermal conductance. Lipid transfer proteins (LTPs) are thought to be involved in the transfer of lipids through the extracellular matrix for the formation of cuticular wax. Using northern analysis, a 6-fold increase of tree tobacco LTP gene transcripts was observed after three drying events, providing further evidence that LTP is involved in cuticle deposition. The simplicity of wax composition and the dramatic wax bloom displayed by tree tobacco make this an excellent species in which to study the relationship between leaf wax deposition and drought tolerance. PMID- 16361525 TI - A circadian rhythm-regulated tomato gene is induced by Arachidonic acid and Phythophthora infestans infection. AB - A cDNA clone of unknown function, DEA1, was isolated from arachidonic acid treated tomato (Solanum lycopersicum) leaves by differential display PCR. The gene, DEA1, is expressed in response to the programmed cell death-inducing arachidonic acid within 8 h following treatment of a tomato leaflet, 16 h prior to the development of visible cell death. DEA1 transcript levels were also affected by the late blight pathogen, Phytophthora infestans. To gain further insight into the transcriptional regulation of DEA1, the promoter region was cloned by inverse PCR and was found to contain putative stress-, signaling-, and circadian-response elements. DEA1 is highly expressed in roots, stems, and leaves, but not in flowers. Leaf expression of DEA1 is regulated by circadian rhythms during long days with the peak occurring at midday and the low point midway through the dark period. During short days, the rhythm is lost and DEA1 expression becomes constitutive. The predicted DEA1 protein has a conserved domain shared by the eight-cysteine motif superfamily of protease inhibitors, alpha-amylase inhibitors, seed storage proteins, and lipid transfer proteins. A DEA1-green fluorescent protein fusion protein localized to the plasma membrane in protoplasts and plasmolysis experiments, suggesting that the native protein is associated with the plasmalemma in intact cells. PMID- 16361526 TI - Identification of a glyphosate-resistant mutant of rice 5-enolpyruvylshikimate 3 phosphate synthase using a directed evolution strategy. AB - 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) is a key enzyme in the shikimate pathway and is targeted by the wide-spectrum herbicide glyphosate. Here, we describe the use of a selection system based on directed evolution to select glyphosate-resistant mutants of EPSPS. Using this system, the rice (Oryza sativa) EPSPS gene, mutagenized by Error-Prone polymerase chain reaction, was introduced into an EPSPS-deficient Escherichia coli strain, AB2829, and transformants were selected on minimal medium by functional complementation. Three mutants with high glyphosate resistance were identified in three independent glyphosate selection experiments. Each mutant contained a C(317)-->T transition within the EPSPS coding sequence, causing a change of proline-106 to leucine (P106L) in the protein sequence. Glyphosate resistance assays indicated a 3-fold increase in glyphosate resistance of E. coli expressing the P106L mutant. Affinity of the P106L mutant for glyphosate and phosphoenolpyruvate was decreased about 70-fold and 4.6-fold, respectively, compared to wild-type EPSPS. Analysis based on a kinetic model demonstrates that the P106L mutant has a high glyphosate resistance while retaining relatively high catalytic efficiency at low phosphoenolpyruvate concentrations. A mathematical model derived from the Michaelis-Menten equation was used to characterize the effect of expression level and selection conditions on kinetic (Ki and Km) variation of the mutants. This prediction suggests that the expression level is an important aspect of the selection system. Furthermore, glyphosate resistance of the P106L mutant was confirmed in transgenic tobacco (Nicotiana tabacum), demonstrating the potential for using the P106L mutant in transgenic crops. PMID- 16361527 TI - Organ-specific expression of glutathione S-transferases and the efficacy of herbicide safeners in Arabidopsis. AB - The functions of plant glutathione S-transferases (GSTs) under normal growth conditions are poorly understood, but their activity as detoxification enzymes has been harnessed in agriculture for selective weed control. Herbicide safeners protect monocot crops from herbicide injury but have little effect on weedy monocot or dicot species. Protection by safeners is associated with expression of herbicide-metabolizing enzymes including GSTs, but the basis for selective action of safeners between monocots and dicots is not known. To address this question we have studied the response of Arabidopsis (Arabidopsis thaliana) to various safeners. Benoxacor, fenclorim, and fluxofenim did not protect Arabidopsis from herbicide injury but did induce RNA expression of the glutathione-conjugate transporters encoded by AtMRP1, AtMRP2, AtMRP3, and AtMRP4. These safeners also induced the organ-specific expression of AtGSTU19 and AtGSTF2, two previously characterized Arabidopsis GSTs from different classes of this enzyme family. RNA hybridization, immunoblot, and reporter gene analyses indicated expression of AtGSTU19 induced by safeners predominated in roots. To test the hypothesis that increased expression of AtGSTU19 would be sufficient to provide tolerance to chloroacetamide herbicides, a chimeric gene was produced containing the open reading frame for this GST driven by a constitutive promoter. Plants containing this transgene had a modest increase in AtGSTU19 protein, predominantly in roots, but this had no effect on tolerance to chloroacetamide herbicides. The localized induction of GSTs by safeners in roots of Arabidopsis may explain why these compounds are unable to provide herbicide tolerance to dicot plant species. PMID- 16361528 TI - Lectin receptor kinases participate in protein-protein interactions to mediate plasma membrane-cell wall adhesions in Arabidopsis. AB - Interactions between plant cell walls and plasma membranes are essential for cells to function properly, but the molecules that mediate the structural continuity between wall and membrane are unknown. Some of these interactions, which are visualized upon tissue plasmolysis in Arabidopsis (Arabidopsis thaliana), are disrupted by the RGD (arginine-glycine-aspartic acid) tripeptide sequence, a characteristic cell adhesion motif in mammals. In planta induced-O (IPI-O) is an RGD-containing protein from the plant pathogen Phytophthora infestans that can disrupt cell wall-plasma membrane adhesions through its RGD motif. To identify peptide sequences that specifically bind the RGD motif of the IPI-O protein and potentially play a role in receptor recognition, we screened a heptamer peptide library displayed in a filamentous phage and selected two peptides acting as inhibitors of the plasma membrane RGD-binding activity of Arabidopsis. Moreover, the two peptides also disrupted cell wall-plasma membrane adhesions. Sequence comparison of the RGD-binding peptides with the Arabidopsis proteome revealed 12 proteins containing amino acid sequences in their extracellular domains common with the two RGD-binding peptides. Eight belong to the receptor-like kinase family, four of which have a lectin-like extracellular domain. The lectin domain of one of these, At5g60300, recognized the RGD motif both in peptides and proteins. These results imply that lectin receptor kinases are involved in protein-protein interactions with RGD-containing proteins as potential ligands, and play a structural and signaling role at the plant cell surfaces. PMID- 16361530 TI - Concordia res parvae crescent. PMID- 16361531 TI - Improved disease-free survival with epirubicin-based chemoendocrine adjuvant therapy compared with tamoxifen alone in one to three node-positive, estrogen receptor-positive, postmenopausal breast cancer patients: results of French Adjuvant Study Group 02 and 07 trials. AB - BACKGROUND: The purpose was to compare disease-free survival (DFS) between epirubicin-based chemoendocrine therapy and tamoxifen alone in one to three node positive (N1-3), estrogen-receptor-positive (ER+), postmenopausal early breast cancer (EBC) patients. PATIENTS AND METHODS: We analyzed, retrospectively, 457 patients randomized in FASG 02 and 07 trials who received: tamoxifen alone (30 mg/day, 3 years); or FEC50 (fluorouracil 500 mg/m2, epirubicin 50 mg/m2, cyclophosphamide 500 mg/m2, six cycles every 21 days) plus tamoxifen started concurrently. Radiotherapy was delivered after the third cycle in FASG 02 trial, and after the sixth in FASG 07 trial. RESULTS: The 9-year DFS rates were 72% with tamoxifen and 84% with FEC50-tamoxifen (P = 0.008). The multivariate analysis showed that pathological tumor size >2 cm was an independent prognostic factor (P = 0.002), and treatment effects remained significantly in favor of chemoendocrine therapy (P = 0.0008). The 9-year overall survival rates were 78% and 86%, respectively (P = 0.11). In the multivariate model, there was a trend in favor of chemoendocrine therapy (P = 0.07). CONCLUSION: The addition of FEC50 adjuvant chemotherapy to tamoxifen significantly improves long-term DFS in N1-3, ER+ and postmenopausal women. Chemoendocrine therapy seems to be more effective than tamoxifen in terms of long-term survival. PMID- 16361533 TI - Intracellular protein degradation and its therapeutic implications. PMID- 16361534 TI - Epithelial to Mesenchymal Transition Tumors: Fallacious or Snail's Pace? PMID- 16361535 TI - Combination analysis of activator protein-1 family members, Sp1 and an activator protein-2alpha-related factor binding to different regions of the urokinase receptor gene in resected colorectal cancers. AB - PURPOSE: Studies on the transactivation of genes via promoter elements have mostly been done on cell lines rather than resected tissues. This, however, is essential to address an in vivo or clinical relevance. We have previously shown tumor-specific binding of Sp1 and an activator protein (AP)-2-related factor to promoter region -152/-135 of the metastasis-related u-PAR gene in 60% of in vivo resected cancer tissues. Cell lines have implicated an additional role, and potential synergism, of an AP-1 region (-190/-171) in u-PAR regulation. This study was done to (a) analyze AP-1 binding to this region in resected tumor and normal tissues, and define subgroups in which it is tumor-specific, and (b) to analyze transcription factor-binding patterns to both promoter motifs in resected tissues, supporting synergism, and draw first prognostic conclusions. EXPERIMENTAL DESIGN: In 103 patients with colorectal cancer, electrophoretic mobility shift assay/supershift analysis for u-PAR promoter region -190/-171 was done in tumors and normal tissues. In 71 patients, region -152/-135 was also analyzed. U-PAR protein was measured by ELISA. RESULTS: Tumor-specific AP-1 binding to region -190/-171 of the u-PAR promoter was found in 40% of patients. Subgroup analysis showed tumor-specific binding for c-Fos in 58%, for c-Jun in 50%, for JunD in 39%, and for Fra-1 in 4% of cases. AP-1 binding correlated significantly with u-PAR protein amounts in both normal and tumor tissues (P<0.001), in contrast to a tumor-specific correlation with u-PAR of the AP-2/Sp1 region. In analyses for both promoter regions, 62% of cancers showed simultaneous binding for AP-1, AP-2, and Sp1, 11% for AP-1 and AP-2, 16% for AP-2 and Sp1, 4% for AP-2 only, 3% for AP-1 only, and 0% for Sp1 only. The binding of AP-1, AP-2, and Sp1 correlated significantly with each other (P<0.001), the combination of AP 1 and AP-2 showing the highest correlation with u-PAR (P=0.008). Preliminary survival analysis indicated a trend for poorer prognosis for binding of all three transcription factors. CONCLUSION: This is the first study differentiating transcription factor-binding to two important u-PAR promoter regions in a large series of resected tumors and normal tissues. The AP-1 site seems to be a less tumor-specific regulator than the Sp1/AP-2 motif. Nevertheless, data corroborate the hypothesis of synergism between both elements in resected tumors. PMID- 16361536 TI - The putative tumor suppressor Cdx2 is overexpressed by human colorectal adenocarcinomas. AB - PURPOSE: The current paradigm suggests that the homeodomain transcription factor Cdx2, which directs the development and maintenance of the intestinal epithelium, is a tumor suppressor in the colon and rectum. Although a cardinal property of tumor suppressors is their inactivation during carcinogenesis, the expression of Cdx2 in colorectal tumors has not been compared with that in normal mucosa. Here, Cdx2 expression and function was quantified in tumors and matched normal mucosa from patients with colorectal cancer. EXPERIMENTAL DESIGN: Cdx2 expression was quantified by reverse transcription-PCR, immunoblot analysis, and immunohistochemistry. Transcriptional activity was explored by quantifying expression of an endogenous downstream target of Cdx2, guanylyl cyclase C (GCC), in tissues by quantitative reverse transcription-PCR and expression of exogenous Cdx2-specific luciferase promoter constructs in epithelial cells isolated from tumors and normal mucosa. RESULTS: Most (>80%) colorectal tumors overexpressed Cdx2 mRNA and protein compared with normal mucosa, with median fold increases of 3.6 and 1.4, respectively (P<0.002). Concomitantly, immunohistochemistry revealed elevated levels of Cdx2 in nuclei of tumor cells compared with normal epithelial cells. Further, tumors exhibited increased expression of GCC compared with normal mucosa. Moreover, cells isolated from tumors overexpressed a Cdx2-specific luciferase promoter construct compared with normal mucosal cells. CONCLUSION: These observations show, for the first time, the structural and functional overexpression of Cdx2 by human colorectal tumors compared with matched normal mucosa. They suggest that loss of Cdx2 expression or transcriptional activity is an infrequent event during tumorigenesis, which does not contribute to molecular mechanisms underlying initiation and progression of most colorectal tumors. PMID- 16361537 TI - Overexpression of platelet-derived growth factor receptor alpha in endothelial cells of hepatocellular carcinoma associated with high metastatic potential. AB - PURPOSE: Little information is available on the heterogeneity of the vascular endothelium in hepatocellular carcinoma. The aim of this study was to identify the differentially expressed genes in tumor endothelial cells from highly metastatic hepatocellular carcinoma. EXPERIMENTAL DESIGN: Magnetic beads conjugated with anti-CD31 antibody were used to isolate vascular endothelial cells from hepatocellular carcinoma xenografts with different metastatic potentials in nude mice. Gene expression profiles for different endothelial cells were compared by use of cDNA microarray. The up-regulated gene was confirmed by reverse transcription-PCR, real-time PCR, and immunohistochemistry. RESULTS: cDNA microarray analysis revealed differential expression patterns in seven genes consistently presented in endothelial cells isolated from hepatocellular carcinoma with different metastatic potentials. Overexpression of platelet derived growth factor receptor alpha was found only in the endothelium of highly metastatic hepatocellular carcinoma, which was confirmed by reverse transcription PCR, real-time PCR, and immunohistochemistry. Oral administration of STI571 (imatinib mesylate or Glivec), a protein tyrosine kinase inhibitor of platelet derived growth factor receptor, combined with s.c. injection of IFN-alpha not only effectively reduced tumor weight (by 81.8%) and microvessel density (by 70.2%) but also inhibited lung metastasis (by 100%). Furthermore, immunohistochemical analysis of platelet-derived growth factor receptor alpha in human hepatocellular carcinoma tissues revealed its correlation with postoperative recurrence, especially in patients without microvessel invasion. CONCLUSIONS: The gene expression of hepatocellular carcinoma vascular endothelium is different between tumors with different metastatic potential. Platelet-derived growth factor receptor alpha, which is overexpressed in endothelium of highly metastatic hepatocellular carcinoma, may serve as a biomarker for predicting metastasis and a therapeutic target for highly metastatic hepatocellular carcinoma. PMID- 16361538 TI - The relationship between global methylation level, loss of heterozygosity, and microsatellite instability in sporadic colorectal cancer. AB - PURPOSE: The relationship between global hypomethylation, chromosomal instability (CIN), and microsatellite instability (MSI) remains unclear in colorectal cancer. The aim of this study was to investigate the relationship between global methylation status, loss of heterozygosity (LOH), and MSI in sporadic colorectal cancer. EXPERIMENTAL DESIGN: We determined global methylation levels in 80 sporadic colorectal cancers, 51 adjacent normal tissues, and 20 normal tissues using the long interspersed nucleotide elements-combined bisulfite restriction analysis method. We also analyzed 80 colorectal cancers for MSI status and LOH at chromosomes 5q21, 8p12-22, 17p13, and 18q21. RESULTS: We identified 14 cases of MSI (17.5%) and 58 cases of LOH (72.5%). LOH was observed more frequently in microsatellite stable (MSS) cancers than in MSI cancers at all loci. Colorectal cancers showed significantly lower global methylation levels than did normal tissues (41.0+/-9.7% versus 54.3+/-6.5%; P<0.001). MSS cancers showed significantly lower global methylation levels when compared with MSI cancers (39.5+/-9.4% versus 48.2+/-8.2%; P=0.003). Tumors with global hypomethylation (with G and -11G>A (transcription start site as position 1), respectively. The complete linkage disequilibrium was found between these two polymorphisms. The frequencies of -27G+/-11A+ genotype and -27G/-11A allele were significantly higher in CML patients than in healthy controls (genotype: 26.36% versus 11.85%, chi2=11.75, P<0.01; allele: 13.57% versus 6.40%, chi2=9.48, P<0.01). Luciferase reporter assays revealed that the promoter with -27G/-11A had significantly lower transcriptional activity and could not be up-regulated after apoptotic stimulations compared with the promoter with -27A/-11G. PDCD5 expression analysis in mononuclear cells derived from CML patients and cell lines with different -27/ 11 genotypes showed consistent results with the reporter assays. CONCLUSIONS: These data suggest that -27G/-11A is associated with reduced PDCD5 promoter activity and increased susceptibility to CML. PMID- 16361543 TI - Magnetic resonance imaging characteristics predict epidermal growth factor receptor amplification status in glioblastoma. AB - PURPOSE: Two clinical-molecular glioblastoma subtypes have been described: "primary" glioblastomas arise de novo in older patients and often overexpress epidermal growth factor receptor (EGFR); "secondary" glioblastomas progress from lower-grade tumors in younger patients and commonly have TP53 mutations. EGFR overexpression correlates in experimental gliomas with increased angiogenesis, edema, and invasion. No radiographic predictors of molecular glioblastoma subtype are known. EXPERIMENTAL DESIGN: We retrospectively reviewed 75 glioblastomas, classified as TP53-mutated (n=11), EGFR-amplified (n=31), or neither (non TP53/non-EGFR; n=33). Four variables were derived from preoperative magnetic resonance imaging: (a) T2/T1, the ratio of T2-bright volume to enclosed T1 enhancing volume; (b) percentage of tumor volume that was necrosis; and (c and d) T1 and T2 border sharpness coefficients (BSC), the rates of change in grayscale intensity of adjacent 0.02-cm2 voxels traversing the anterior, posterior, and lateral borders on T1-enhanced and T2 images. RESULTS AND CONCLUSIONS: Mean T2/T1 was 4.7 for EGFR-amplified glioblastomas, greater than that of TP53-mutated glioblastomas (2.3) or non-TP53/non-EGFR glioblastomas (2.6; P<0.00005). All four tumors with T2/T1>7.2 were EGFR-amplified; 0 of 15 with T2/T1<4.7 underwent gross total resection. The mean T2 BSC of EGFR-amplified glioblastomas was 33.7, less sharp (P<0.0000005) than TP53-mutated (72.2) and non-TP53/non-EGFR glioblastomas (81.2). All 15 glioblastomas with T2 BSC<30.8 were EGFR-amplified. Percentage necrosis and T1 BSC did not differ between glioblastoma subtypes. The increased T2/T1 ratio and decreased T2 BSC in EGFR-overexpressing tumors are the first radiographic distinctions described between glioblastoma molecular subtypes. These findings may reflect increased angiogenesis, edema, and/or invasion in EGFR overexpressing tumors. PMID- 16361544 TI - Importance of P-cadherin, beta-catenin, and Wnt5a/frizzled for progression of melanocytic tumors and prognosis in cutaneous melanoma. AB - PURPOSE: It has been proposed that melanoma cells shift from E-cadherin to N cadherin expression during tumor development, and recent gene profiling has shown increased expression of Wnt5a/Frizzled in aggressive melanomas possibly by interactions with beta-catenin. We therefore wanted to investigate the role of cadherin subtypes, beta-catenin, and Wnt5a/Frizzled in melanocytic tumors, with focus on prognosis in nodular melanomas. EXPERIMENTAL DESIGN: The immunohistochemical expression of E-cadherin, N-cadherin, P-cadherin, beta catenin, and Wnt5a/Frizzled was examined using tissue microarrays of 312 melanocytic tumors. RESULTS: Cytoplasmic expression of P-cadherin was associated with increasing tumor thickness (P=0.005) and level of invasion (P=0.019), whereas membranous staining was associated with thinner (P=0.012) and more superficial (P=0.018) tumors. Increased cytoplasmic P-cadherin was associated with reduced survival (P=0.047). Lack of nuclear beta-catenin expression was related to increased tumor thickness (P=0.002) and poor patient survival in univariate (P=0.0072) and multivariate (P=0.004) analyses. Membranous expression of N-cadherin was significantly increased from primary tumors to metastatic lesions, whereas E-cadherin staining tended to be decreased. Wnt5a and its receptor Frizzled were highly coexpressed, and nuclear expression of both markers was significantly reduced from benign nevi to melanomas, with a shift from nuclear to cytoplasmic expression in malignant tumors. In addition, Wnt5a expression was significantly associated with nuclear beta-catenin expression. CONCLUSIONS: Alterations in the expression and subcellular localization of cell adhesion markers are important in the development and progression of melanocytic tumors, and strong cytoplasmic P-cadherin expression and loss of nuclear beta catenin staining were associated with aggressive melanoma behavior and reduced patient survival. PMID- 16361545 TI - Tumor beta-1,4-galactosyltransferase IV overexpression is closely associated with colorectal cancer metastasis and poor prognosis. AB - PURPOSE: To elucidate the significance of beta-1,4-galactosyltransferase IV (beta 1,4-GT-IV) in the clinical presentation and prognostication of colorectal cancer. EXPERIMENTAL DESIGN: Tissue lysates from paired tumor and nontumor tissues of a colon cancer patient were labeled separately with fluorescent dyes Cy5 and Cy3 for two-dimensional difference in-gel electrophoresis. Subsequent matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and immunoblot analyses identified a down-regulated level of beta-1,4-GT-IV in the tumor tissue. In the follow-up study, paired tissue lysates were obtained from 100 colorectal cancer patients with immunoblot analyses done to compare the levels of beta-1,4 GT-IV expression in these patients. RESULTS: Of 100 colorectal patients studied, 48% had down-regulated expression of beta-1,4-GT-IV in the tumor tissue but 28% of patients exhibited elevated beta-1,4-GT-IV levels. Increased beta-1,4-GT-IV in the tumor tissue was significantly coexistent with raised serum level of CA-199 and the presence of tumor metastasis (P=0.006 and P<0.001, respectively) but was independent of age and gender of patient, tumor site, tumor size, serum level of carcinoembryonic antigen, grade of tumor cell differentiation, and depth of tumor invasion. The results of logistic regression analyses suggested that tumor beta 1,4-GT-IV overexpression and tumor invasion, but not other patient variables such as tumor size and serum levels of carcinoembryonic antigen and CA19-9, were significantly correlated with the occurrence of metastases (P<0.05). In a multivariate regression analysis, the patient group with tumor beta-1,4-GT-IV overexpression strongly predicted for tumor metastasis (odds ratio, 10.009; 95% confidence interval, 2.992-33.484; P<0.001). Likewise, tumor beta-1,4-GT-IV overexpression was significantly associated with poor overall survival (P<0.01). By Cox regression analysis, this association remained significant even after adjustment for tumor metastasis (P=0.048). CONCLUSION: Increased beta-1,4-GT-IV expression in tumor tissue was strongly associated with tumor metastases and poor prognosis in colorectal cancer. PMID- 16361546 TI - Tumor gene expression and prognosis in breast cancer patients with 10 or more positive lymph nodes. AB - PURPOSE: This study, along with two others, was done to develop the 21-gene Recurrence Score assay (Oncotype DX) that was validated in a subsequent independent study and is used to aid decision making about chemotherapy in estrogen receptor (ER)-positive, node-negative breast cancer patients. EXPERIMENTAL DESIGN: Patients with >or=10 nodes diagnosed from 1979 to 1999 were identified. RNA was extracted from paraffin blocks, and expression of 203 candidate genes was quantified using reverse transcription-PCR (RT-PCR). RESULTS: Seventy-eight patients were studied. As of August 2002, 77% of patients had distant recurrence or breast cancer death. Univariate Cox analysis of clinical and immunohistochemistry variables indicated that HER2/immunohistochemistry, number of involved nodes, progesterone receptor (PR)/immunohistochemistry (% cells), and ER/immunohistochemistry (% cells) were significantly associated with distant recurrence-free survival (DRFS). Univariate Cox analysis identified 22 genes associated with DRFS. Higher expression correlated with shorter DRFS for the HER2 adaptor GRB7 and the macrophage marker CD68. Higher expression correlated with longer DRFS for tumor protein p53-binding protein 2 (TP53BP2) and the ER axis genes PR and Bcl2. Multivariate methods, including stepwise variable selection and bootstrap resampling of the Cox proportional hazards regression model, identified several genes, including TP53BP2 and Bcl2, as significant predictors of DRFS. CONCLUSION: Tumor gene expression profiles of archival tissues, some more than 20 years old, provide significant information about risk of distant recurrence even among patients with 10 or more nodes. PMID- 16361547 TI - Mcl-1, vascular endothelial growth factor-R2, and 14-3-3sigma expression might predict primary response against radiotherapy and chemotherapy in patients with locally advanced squamous cell carcinomas of the head and neck. AB - PURPOSE: This study was done to explore whether the expression of a selected set of proteins could predict primary response to radiotherapy or concomitant radiotherapy and chemotherapy in patients with advanced head and neck cancer. EXPERIMENTAL DESIGN: Forty-three pretreatment tumor biopsies were taken during diagnostic panendoscopy and examined for Mcl-1, vascular endothelial growth factor (VEGF)-R2, CD9, and 14-3-3sigma expression by immunohistochemistry. Forty three patients underwent primary radiotherapy, of which, 29 patients received concomitant chemotherapy (low dose daily cisplatin, mitomycin C bolus). The primary end-point was locoregional tumor control 6 months after completion of radiotherapy. Mcl-1, VEGF-R2, CD9, and 14-3-3sigma expression were correlated with patients' primary response to radiotherapy and chemotherapy and with established clinicopathologic variables. RESULTS: Thirty complete and 13 partial responses were observed in our patient group. High expression levels of Mcl-1 (P=0.021), VEGF-R2 (P=0.032), and 14-3-3sigma (P=0.013), but not of CD9, in tumor biopsies was correlated with complete response. Overexpression of at least two of the three aforementioned proteins in pretreatment biopsies predicted-with a likelihood of 80%-whether a patient would achieve complete response to radiotherapy and chemotherapy. However, if only one of these proteins is overexpressed, there is a likelihood of 84.6% that this patient would not completely respond to therapy. CONCLUSION: Determining the expression levels of Mcl-1, VEGF-R2, and 14-3-3sigma may be helpful in predicting the early clinical response in head and neck tumor patients receiving primary radiotherapy and chemotherapy and may further allow a pretherapeutic selection of patients. PMID- 16361548 TI - Effect of epidermal growth factor receptor expression level on survival in patients with epithelial ovarian cancer. AB - BACKGROUND: Several lines of laboratory evidence support the epidermal growth factor receptor (EGFR) as an adverse prognostic indicator in ovarian cancers. However, different methods of immunohistochemical assessment have yielded conflicting results. Here, we sought to determine the prognostic value of EGFR in ovarian cancer using a novel method of compartmentalized in situ protein analysis. METHODS: A tissue array composed of 150 advanced-stage ovarian cancers uniformly treated, with surgical debulking followed by platinum-paclitaxel combination chemotherapy, was constructed. For evaluation of EGFR protein expression, we used an immunofluorescence-based method of automated in situ quantitative measurement of protein analysis (AQUA). RESULTS: Mean follow-up time for the entire cohort was 34.4 months. Eighty-one of 150 cases had sufficient tissue for AQUA analysis. High tumor EGFR expression was associated with poor outcome for overall survival (P=0.0001) and disease-free survival (P=0.0005) at 3 years. In multivariable analysis, adjusting for well-characterized prognostic variables, EGFR expression status was the most significant prognostic factor for disease-free and overall survival. CONCLUSION: The conflicting results in the literature regarding the prognostic value of EGFR may be due to the technical difficulties inherent in assessing EGFR with immunocytochemistry. In the present study, we show that measurement of EGFR protein levels in ovarian cancer using AQUA is feasible and can give important prognostic information. PMID- 16361549 TI - High serum concentration of YKL-40 is associated with short survival in patients with acute myeloid leukemia. AB - PURPOSE: YKL-40 is secreted by cancer cells, macrophages, and neutrophils. It may be a growth or differentiation factor, play a role in angiogenesis, or protect against apoptosis. High serum YKL-40 is associated with poor prognosis in solid carcinomas. The aim was to examine serum YKL-40 in patients with acute myeloid leukemia (AML). EXPERIMENTAL DESIGN: YKL-40 was measured by ELISA in serum from 77 patients recently diagnosed with AML before and during the first month of chemotherapy. RESULTS: Forty (52%) of the AML patients had elevated serum YKL-40 (compared with age-matched healthy subjects) and their survival was shorter than in patients with normal serum YKL-40 (median, 128 days; interquartile range, 18 629 days versus 386 days; interquartile range, 180-901; P=0.018 Mann-Whitney test). Univariate analysis of serum YKL-40 (logarithmically transformed and treated as a continuous covariate) showed significant association with survival within the first month after start of chemotherapy [hazard ratio (HR), 1.7; 95% confidence interval (CI), 1.2-2.4; P=0.002], first 12 months (HR, 1.6; 95% CI, 1.2-2.0; P=0.0002), and overall survival (HR, 1.3; 95% CI, 1.1-1.6; P=0.003). Multivariate Cox analysis showed that serum YKL-40 was an independent prognostic variable for survival (first month: HR, 1.7; P=0.011; 12 months: HR, 1.6; P=0.0002; overall survival: HR, 1.4; P=0.002). High serum YKL-40 at start of chemotherapy was a risk factor for pneumonia within the first month, and serum YKL-40 increased (P=0.002) at time of pneumonia and was unchanged in patients without infections. CONCLUSIONS: Serum YKL-40 is a prognostic biomarker of survival in AML patients. Its role in AML and infections needs to be determined. PMID- 16361550 TI - Inclusion of vasculature-related variables in the Dukes staging system of colon cancer. AB - PURPOSE: The Dukes stage is used to stratify colorectal cancer patients into groups of different prognosis and need of adjuvant radiotherapy and chemotherapy. However, approximately 80% of patients with Dukes stage C colorectal cancer receive cytotoxic therapy without any expected benefit, for such patients would either not relapse without adjuvant therapy or they would inevitably do so because of tumor resistance to the available regimens. On the other hand, as 20% of Dukes stage B patients would relapse after surgery, adjuvant therapy could improve their survival. Improvement of the Dukes stage predictive accuracy is necessary to better assign patients for adjuvant therapies, especially nowadays when antiangiogenic agents are being incorporated in the clinical practice. PATIENTS AND METHODS: In this study, we examined the prognostic role of Dukes staging system in parallel with three vasculature-related variables (vascular invasion, tumor angiogenic activity, and vascular survival ability) in a series of 130 stage B/C patients with colorectal cancer treated with surgery alone (without adjuvant radiotherapy or chemotherapy). RESULTS: Inclusion of vasculature-related variables in the Dukes staging system significantly improved the prognostic categorization of patients, identifying subgroups of B-stage and C stage patients with an up to 40% and 60% 5-year survival difference, respectively. CONCLUSIONS: Preliminary results show that the prognostic value of Dukes staging system is significantly improved after taking into account vasculature-related variables, which may be useful in stratifying patients for adjuvant therapies, highlighting also subgroups that may benefit the most from antiangiogenic agents. PMID- 16361551 TI - Expression of multidrug resistance-associated proteins predicts prognosis in childhood and adult acute lymphoblastic leukemia. AB - PURPOSE: Patients with acute lymphoblastic leukemia (ALL) are treated with a variety of chemotherapeutic drugs, which can be transported by six multidrug resistance-associated proteins (MRP). These MRPs have strongly overlapping functional activities. The aim of this study was to investigate the expression levels of MRP1 to MRP6 and study their effect on prognosis. EXPERIMENTAL DESIGN: The mRNA expression levels of MRP1 to MRP6 were analyzed by quantitative real time PCR in leukemic blasts of 105 de novo ALL patients (adults, n=49; children, n=56) including 70% B-lineage and 30% T-lineage ALL patients. RESULTS: Adults showed a higher expressions of MRP1 (P=0.008), MRP2 (P=0.026), and MRP3 (P=0.039) than children. Interestingly, this difference disappeared when patients were categorized based on clinical outcome. Relapsed patients showed a higher expression of all MRP genes, except MRP4. For the total group of ALL patients, the expressions of MRP1, MRP2, MRP3, MRP5, and MRP6 predicted relapse. Moreover, high expression of all MRP genes, except MRP4, was associated with a reduced relapse-free survival in children and adults (MRP1, P=0.005; MRP2, P=0.008; MRP3, P=0.001; MRP5, P=0.016; MRP6, P=0.037). CONCLUSIONS: The present study shows that a subset of ALL patients with high MRP expression has an unfavorable prognosis independently of age. PMID- 16361552 TI - Time to detectable metastatic disease in patients with rising prostate-specific antigen values following surgery or radiation therapy. AB - PURPOSE: To determine factors associated with the development of radiographic metastatic progression for patients with recurrent prostate cancer following surgery and/or radiation therapy with prostate-specific antigen (PSA) doubling times of <12 months. EXPERIMENTAL DESIGN: One hundred and forty-eight patients with rising PSA values after primary therapy and a PSA doubling time of <12 months enrolled on clinical protocols were followed and monitored at protocol specified intervals with examinations, PSA determinations, and imaging studies that included a computed tomography or magnetic resonance imaging and bone scan until metastases were detected. Metastasis-free survival was estimated using the Kaplan-Meier method and factors predictive of progression-free survival were estimated using the proportional hazards model. A nomogram based on the Cox model was constructed. RESULTS: Metastatic events were documented in 74% (110 of 148) of patients during the follow-up period. The median progression-free survival was 19 months, with 3- and 5-year metastatic progression-free survival of 32% and 16%, respectively. T stage (P=0.07) and Gleason grade (P=0.006) at the time of diagnosis, PSA values at the time of protocol entry (P<0.001), and PSA doubling time (P<0.001) were associated with progression in univariate analysis. These were combined into a nomogram to assess risk for an individual patient. CONCLUSIONS: Tumor characteristics at the time of diagnosis, PSA doubling time following relapse, and the PSA value at the time of the protocol are predictive of metastatic progression. Because the PSA value at the time of monitoring was predictive, early treatment to prevent metastatic progression is favored. PMID- 16361553 TI - Circulating aminopeptidase N/CD13 is an independent prognostic factor in patients with non-small cell lung cancer. AB - PURPOSE: Aminopeptidase N, also known as CD13, has important roles in tumor metastasis and angiogenesis. Its expression in tumor tissue has been reported to be associated with poor prognosis. However, the clinical significance of circulating aminopeptidase N/CD13 in patients with solid tumors is unknown. We previously developed an aminopeptidase N/CD13-specific monoclonal antibody (mAb) MH8-11, which inhibits cell motility and angiogenesis in vitro. The aim of this study was to evaluate the clinical significance of circulating aminopeptidase N/CD13 protein detected by mAb MH8-11 in patients with non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: We used electrochemiluminescence immunoassay with mAb MH8-11 to determine circulating aminopeptidase N/CD13 levels in 90 healthy volunteers and 90 patients with NSCLC. Circulating aminopeptidase N/CD13 levels were measured in sera taken before treatment and evaluated for a relationship with clinical outcomes. RESULTS: A significant correlation was found between tumor progression and serum aminopeptidase N/CD13 concentrations (r=0.23, P=0.029). High serum aminopeptidase N/CD13 levels (n=17) were associated with advanced stage (P=0.004) or poor performance status (P=0.001). The overall survival rate for patients with high serum aminopeptidase N/CD13 levels (n=17) was significantly less than that of patients with low serum aminopeptidase N/CD13 levels (n=73, P<0.0001). In a multivariate survival analysis in patients with NSCLC, serum aminopeptidase N/CD13 levels had an independent influence on survival (relative risk, 4.1; 95% confidence interval, 1.9-8.8). CONCLUSIONS: Our data suggest that a high level of circulating aminopeptidase N/CD13 at diagnosis is an independent prognostic factor in patients with NSCLC. PMID- 16361554 TI - Phase I/II study of 19-nor-1alpha-25-dihydroxyvitamin D2 (paricalcitol) in advanced, androgen-insensitive prostate cancer. AB - PURPOSE: We assessed the safety and efficacy of the vitamin D analogue, 19-nor 1alpha-25-dihydroxyvitamin D2 (paricalcitol), in patients with androgen independent prostate cancer. EXPERIMENTAL DESIGN: Patients received paricalcitol i.v. three times per week on an escalating dose of 5 to 25 microg (3-15 microg/m2). The primary end point was prostate-specific antigen (PSA) response. Secondary end points were characterization of toxicity in this population, changes in serum parathyroid hormone (PTH), and survival. RESULTS: A total of 18 patients were enrolled. No patient showed a sustained 50% drop in serum PSA, despite several large declines in PSA (e.g., 1,300 ng/mL). Paricalcitol was well tolerated. One instance of significant hypercalcemia, a serum calcium of 14.3 mg/dL, was observed at the highest dose (25 microg). At entry into the study, seven (41%) of the patients had elevated serum levels of PTH, which were significantly reduced by paricalcitol. Higher levels of serum PTH at study entry were significantly and negatively associated with survival (P<0.01). CONCLUSION: No objective responses were seen in the primary end point. However, elevated serum levels of PTH, a common feature of advanced prostate cancer, were reduced by paricalcitol. Because elevated PTH is associated with increased cardiovascular and skeletal morbidity, including an increased risk for pathologic fracture, further evaluation of paricalcitol in the reduction of skeletal morbidity in advanced prostate cancer is warranted. PMID- 16361555 TI - Epithelial versus mesenchymal phenotype determines in vitro sensitivity and predicts clinical activity of erlotinib in lung cancer patients. AB - Significant improvements in the outcome of non-small cell lung carcinoma (NSCLC) have been reported in patients treated with the epidermal growth factor receptor (EGFR) inhibitor, erlotinib. To discover biomarkers for the enrichment of patients who might benefit from treatment, a pharmacogenomic approach was used to identify gene signatures that may predict erlotinib activity using in vitro model systems. Erlotinib sensitivity in a panel of 42 NSCLC cell lines was determined by EGFR-mediated proliferative potential, EGFR mutations, and/or EGFR gene amplification, thus supporting an underlying biological mechanism of receptor activation. A strong multigene signature indicative of an epithelial to mesenchymal transition (EMT) was identified as a determinant of insensitivity to erlotinib through both supervised and unsupervised gene expression approaches. This observation was further supported by expression analysis of classic EMT marker proteins, including E-cadherin and vimentin. To investigate the clinical relevance of these findings, we examined expression of the epithelial marker E cadherin by immunohistochemistry on primary tumor samples from subjects enrolled in a randomized NSCLC clinical trial in which erlotinib in combination with chemotherapy previously failed to show clinical activity. The majority (75%) of the 87 subjects tested showed strong E-cadherin staining and exhibited a significantly longer time to progression (hazard ratio, 0.37; log rank P=0.0028) and a nonsignificant trend toward longer survival with erlotinib plus chemotherapy treatment versus chemotherapy alone. These data support a potential role for EMT as a determinant of EGFR activity in NSCLC tumor cells and E cadherin expression as a novel biomarker predicting clinical activity of the EGFR inhibitor erlotinib in NSCLC patients. PMID- 16361556 TI - Methylation of the DPYD promoter: an alternative mechanism for dihydropyrimidine dehydrogenase deficiency in cancer patients. AB - PURPOSE: Dihydropyrimidine dehydrogenase (DPD) deficiency, a known pharmacogenetic syndrome associated with 5-fluorouracil (5-FU) toxicity, has been detected in 3% to 5% of the population. Genotypic studies have identified >32 sequence variants in the DPYD gene; however, in a number of cases, sequence variants could not explain the molecular basis of DPD deficiency. Recent studies in cell lines indicate that hypermethylation of the DPYD promoter might down regulate DPD expression. The current study investigates the role of methylation in cancer patients with an unexplained molecular basis of DPD deficiency. EXPERIMENTAL DESIGN: DPD deficiency was identified phenotypically by both enzyme assay and uracil breath test, and genotypically by denaturing high-performance liquid chromatography. The methylation status was evaluated in PCR products (209 bp) of bisulfite-modified DPYD promoter, using a novel denaturing high performance liquid chromatography method that distinguishes between methylated and unmethylated alleles. Clinical samples included five volunteers with normal DPD enzyme activity, five DPD-deficient volunteers, and five DPD-deficient cancer patients with a history of 5-FU toxicity. RESULTS: No evidence of methylation was detected in samples from volunteers with normal DPD. Methylation was detected in five of five DPD-deficient volunteers and in three of five of the DPD-deficient cancer patient samples. Of note, one of the two samples from patients with DPD deficient cancer with no evidence of methylation had the mutation DPYD*2A, whereas the other had DPYD*13. DISCUSSION: Methylation of the DPYD promoter region is associated with down-regulation of DPD activity in clinical samples and should be considered as a potentially important regulatory mechanism of DPD activity and basis for 5-FU toxicity in cancer patients. PMID- 16361557 TI - Serum free light chain analysis and urine immunofixation electrophoresis in patients with multiple myeloma. AB - PURPOSE: Retrospective studies have shown that immunoassays measuring free light chains (FLC) in serum are useful for diagnosis and monitoring of multiple myeloma. This study prospectively evaluates the use of FLC assays and, for the first time, investigates the relationship between serum FLC concentrations and the presence and detectability of Bence Jones (BJ) proteins in the urine. PATIENTS AND METHODS: Three hundred seventy-eight paired samples of serum and urine were tested from 82 patients during the course of their disease. The sensitivities of serum FLC analysis and urine immunofixation electrophoresis (IFE) in detecting monoclonal FLC were compared. Serum FLC concentrations required for producing BJ proteins detected by IFE were determined. RESULTS: Abnormal FLC were present in 54% of serum samples compared with 25% by urine tests. In abnormal serum samples for kappa or lambda, the sensitivity of IFE to detect the respective BJ proteins in urine were 51% and 35% and the median serum FLC concentrations required to produce detectable BJ proteins were 113 and 278 mg/L. Renal excretions of monoclonal FLC increased with serum concentrations, but excretions significantly decreased at high serum concentrations combined with renal dysfunction. CONCLUSION: Serum FLC assays are significantly more sensitive for detecting monoclonal FLC than urine IFE analysis. They also have the advantage of FLC quantification and are more reliable for monitoring disease course and response to treatment. PMID- 16361558 TI - Feasibility and tolerability of sequential doxorubicin/paclitaxel followed by cyclophosphamide, methotrexate, and fluorouracil and its effects on tumor response as preoperative therapy. AB - PURPOSE: The European Cooperative Trial in Operable breast cancer (ECTO) randomly tested whether efficacy of adjuvant doxorubicin followed by i.v. cyclophosphamide, methotrexate, and fluorouracil (CMF; doxorubicin-->CMF, arm A) could be improved by adding paclitaxel (doxorubicin/paclitaxel-->CMF) as adjuvant (arm B) or primary systemic therapy (PST, arm C). We report here feasibility, tolerability, locoregional antitumor activity, and breast conservation rate. METHODS: A total of 1,355 women entered the study. Feasibility and safety were compared in arm A versus arms B plus C. Surgical findings were compared in arms A plus B versus arm C. RESULTS: Grade 3 or 4 National Cancer Institute toxicities were low (<5%) in all arms. Neuropathy was more frequent in the paclitaxel containing arms (grade 2, 20.5% versus 5.0%; grade 3, 1.3% versus 0.2%). At 31 months of follow-up, asymptomatic drop of left ventricular ejection fraction was similar in all arms, whereas symptomatic cardiotoxicity was recorded in three patients (0.5%) in A and in three patients (0.3%) in B plus C. PST induced clinical complete plus partial remission in 78%, with an in-breast pathologic complete response rate of 23% and an in-breast plus axilla pathologic complete response rate of 20%. In the multivariate analysis, only estrogen receptor (ER) status was significantly associated with pathologic complete response (odds ratio for ER negative, 5.77; 95% confidence interval, 3.49-9.52; P<0.0001). PTS induced a significant axillary downstaging (P<0.001), and breast sparing surgery was feasible in 65% versus 34% (P<0.001). CONCLUSIONS: Doxorubicin/paclitaxel-->CMF is feasible, safe, and well tolerated. Given as PST, it is markedly active, allowing for breast-sparing surgery in a large fraction of patients. PMID- 16361559 TI - Effect of exemestane on tamoxifen pharmacokinetics in postmenopausal women treated for breast cancer. AB - PURPOSE: Rodent models of human breast cancer suggest that the combination of the steroidal aromatase inhibitor exemestane with tamoxifen may have additive activity. Clinical trials combining tamoxifen with letrozole or anastrazole have shown minor pharmacokinetic drug interactions. We did an open-label crossover clinical trial of the effect of exemestane on tamoxifen pharmacokinetics. DESIGN: Thirty-two postmenopausal women who were clinically disease-free following primary treatments for breast cancer receiving tamoxifen for at least 3 months were studied. Blood was collected for pharmacokinetic analysis after at least 4 months of receiving 20 mg tamoxifen daily. Subjects then began 8 weeks of oral exemestane (25 mg daily), followed by another set of blood samples. RESULTS: There were no serious toxicities noted when the two drugs were combined. There was no significant effect of exemestane on the area under the plasma concentration versus time curve (AUC) of tamoxifen at steady state before [3.04 mg h/L; 90% confidence interval (90% CI), 2.71-3.44] and during exemestane treatment (3.05 mg h/L; 90% CI, 2.72-3.41). There were no significant changes in the formation of primary tamoxifen metabolites. Oral clearance of exemestane averaged 602 L/h based on an average plasma exemestane AUC of 41.5 microg h/L (90% CI, 36.7-62.6). Plasma concentrations of estradiol, estrone, and estrone sulfate decreased when exemestane was begun; estradiol concentrations consistently decreased below the limit of quantitation. CONCLUSIONS: There is no pharmacokinetic interaction between tamoxifen and exemestane. No modification in the standard regimen of either drug seems to be indicated if they are used in combination. The combination of the two drugs was well tolerated during the 8 week evaluation period. PMID- 16361560 TI - Phase I and pharmacokinetic study of sequences of the rebeccamycin analogue NSC 655649 and cisplatin in patients with advanced solid tumors. AB - PURPOSE: To evaluate the feasibility of administering NSC 655649, a water-soluble rebeccamycin analogue that inhibits both topoisomerases I and II, in combination with cisplatin (CDDP) in adults with solid malignancies. Major toxicologic and pharmacologic differences between the two sequences of drug administration were also assessed. EXPERIMENTAL DESIGN: NSC 655649 was administered as a 60-minute i.v. infusion; CDDP was given i.v. before or after NSC 655649 on day 1. Each patient was treated with alternating drug sequences every 3 weeks; doses of each drug were escalated in separate cohorts of new patients. Sequential dose escalation of NSC 655649 or CDDP resulted in three dosage permutations of NSC 655649/CDDP: 440/50, 550/50, and 440/75 mg/m2. After the maximum tolerated dose level was determined, the feasibility of using granulocyte colony-stimulating factor to permit further dose escalation was explored. RESULTS: Twenty patients were treated with 70 courses of NSC 655649/CDDP. Myelosuppression was the principal toxicity. The incidence of severe neutropenia, often associated with severe thrombocytopenia, was unacceptably high in minimally pretreated patients at the NSC 655649/CDDP dose level of 550/50 mg/m2 without and with granulocyte colony-stimulating factor. Major pharmacokinetic interactions between NSC 655649 and CDDP were not apparent. No relevant sequence-dependent differences in toxicity or pharmacokinetic variables occurred. Three patients had partial responses. CONCLUSIONS: NSC 655649 and CDDP were well tolerated by minimally pretreated subjects at 440 and 50 mg/m2, respectively. Neither pharmacokinetic interactions between the agents nor sequence-dependent toxicologic or pharmacokinetic effects were apparent. The tolerance and preliminary activity observed with this combination suggest that disease-directed evaluations of the regimen are warranted. PMID- 16361561 TI - Pigment epithelium-derived factor gene therapy inhibits human pancreatic cancer in mice. AB - PURPOSE: Pigment epithelium-derived factor (PEDF), which has recently been shown to be the most potent inhibitor of angiogenesis in the mammalian eye, is also expressed in the pancreas. Previously, we have screened the expression of PEDF by immunohistochemical analysis and showed that low expression of PEDF is associated with increased risk of hepatic metastasis and short survival. The purpose of this study was to investigate whether PEDF gene is a potent tumor suppressor and a potential candidate for cancer gene therapy. EXPERIMENTAL DESIGN: We investigated both in vitro and in vivo growth characteristics of human pancreatic adenocarcinoma cell lines that were stably transfected to overexpress human PEDF and therapeutic effects of lentivirus-based vectors expressing PEDF on tumor growth in murine s.c. tumor model. RESULTS: We discovered that cells secreted PEDF protein in the media and this exhibited strong inhibitory effects on proliferation and migration of human umbilical vein endothelial cells. The size of PEDF-overexpressing pancreatic adenocarcinoma tumors was significantly smaller than that of control tumors in s.c. tumor models. Moreover, the growth of PEDF overexpressing pancreatic adenocarcinoma cells was significantly suppressed in comparison with control cells in peritoneal metastasis models. In gene transfer models, intratumoral injection of a lentivirus vector encoding PEDF (LV-PEDF) caused significant inhibition of tumor growth. The antitumor effect observed after treatment with LV-PEDF was associated with decreased microvessel density in tumors. CONCLUSION: Our data suggest that PEDF may exert a biological effect on tumor angiogenesis and PEDF gene therapy may provide a new approach for treatment of pancreatic adenocarcinoma. PMID- 16361562 TI - Neamine inhibits xenografic human tumor growth and angiogenesis in athymic mice. AB - PURPOSE: We have previously shown that the aminoglycoside antibiotic neomycin blocks the nuclear translocation of angiogenin and inhibits its angiogenic activity. However, neomycin has not been considered as a favorable drug candidate for clinical development because of its known nephrotoxicity and ototoxicity. The aim of this study is to determine whether neamine, a nontoxic derivative of neomycin, possesses antitumor activity. EXPERIMENTAL DESIGN: The effect of neamine on the nuclear translocation of angiogenin was examined by means of immunofluorescence and Western blotting. The antitumor activity of neamine was determined with three different animal models. RESULTS: Neamine effectively blocked the nuclear translocation of angiogenin in endothelial cells and inhibited angiogenin-induced cell proliferation. It inhibited the establishment of human tumor xenografts in athymic mice in both ectopic and orthotopic tumor models. It also inhibited the progression of established human tumor transplants, whereas the structurally related antibiotic paromomycin had no effect. Immunohistochemical staining showed that both angiogenesis and cancer cell proliferation are inhibited by neamine. CONCLUSION: These results suggest that the nontoxic aminoglycoside antibiotic neamine is an effective inhibitor of nuclear translocation of angiogenin and may serve as an inhibitor for angiogenin induced angiogenesis and cancer progression. PMID- 16361563 TI - Overexpression of stefin A in human esophageal squamous cell carcinoma cells inhibits tumor cell growth, angiogenesis, invasion, and metastasis. AB - PURPOSE: Evidence is accumulating that an inverse correlation exists between stefin A level and malignant progression. The aim of this study is to investigate the role of stefin A in human esophageal squamous cell carcinoma cells and to evaluate the possibility of stefin A for cancer therapy. EXPERIMENTAL DESIGN: We stably transfected stefin A cDNA into human EC9706 or KYSE150 esophageal squamous cell carcinoma cells. Subsequently, we evaluated the effect of stefin A overexpression on cell growth, cathepsin B activity, cell motility and invasion, tumor growth, and metastasis. Immunoanalysis was done to assess the expression of factor VIII and to support the localization of stefin A and cathepsin B. We also evaluated the effect of CA074Me, a selective membrane-permeant cathepsin B inhibitor. RESULTS: Both transfection of stefin A and treatment with 10 micromol/L CA074Me significantly reduced cathepsin B activity and inhibited the Matrigel invasion. Combination of both further reduced cathepsin B activity and inhibited the Matrigel invasion. Overexpression of stefin A delayed the in vitro and in vivo growth of cells and significantly inhibited lung metastasis compared with 50% of lung metastasis in xenograft mice from EC9706 or empty vector cells. Transfection with stefin A showed a dramatic reduction of factor VIII staining in the tumors of xenograft mice. CONCLUSIONS: Our data strongly indicate that stefin A plays an important role in the growth, angiogenesis, invasion, and metastasis of human esophageal squamous cell carcinoma cells and suggest that stefin A may be useful in cancer therapy. PMID- 16361564 TI - Natural killer cells license dendritic cell cross-presentation of B lymphoma cell -associated antigens. AB - PURPOSE: Presentation of exogenous antigen by MHC class I molecules, or cross presentation, is a property of dendritic cells, which is considered crucial for the priming of cytotoxic T-cell response to tumor antigens. However, the precise mechanisms of this process are not fully understood. EXPERIMENTAL DESIGN AND RESULTS: We show here in a human in vitro system, using B lymphoma cells as a tumor model, that the cross-presentation of cell-associated antigens to T cells by dendritic cells requires "help" from natural killer cells. When autologous dendritic cells that had taken up apoptotic B lymphoma cells and induced to a fully mature state were used to stimulate nonadherent cells of peripheral blood mononuclear cells from healthy donors, they induced strong cytotoxicity against B lymphoma cells in a HLA-A0201-restricted manner. The cells failed to induce cytotoxicity, however, when purified T cells were used as effector cells. Depletion of CD56+ cells, but not CD14+ or CD19+ cells, abrogated the cytotoxicity of nonadherent cells, showing that the help was provided by natural killer cells. Further, when natural killer cells were present in the cultures, a strong and persistent production of interleukin-18, but not interleukin-12 and interleukin-15, was observed. Blocking interleukin-18 significantly reduced the cytotoxicity of nonadherent cells against B lymphoma cells. CONCLUSIONS: These results suggest that capture of tumor cells and a full maturation status of dendritic cells are not sufficient to cross-prime CD8 T cells. Effective cross priming requires further activation of dendritic cells by natural killer cells and an abundant production of interleukin-18, which, along with other yet undefined mechanisms, contribute to the generation of CTL response against B-cell lymphoma. PMID- 16361565 TI - Antitumor efficacy of capecitabine and celecoxib in irradiated and lead-shielded, contralateral human BxPC-3 pancreatic cancer xenografts: clinical implications of abscopal effects. AB - PURPOSE: X-ray therapy (XRT) remains one of the major modalities used to treat patients diagnosed with locally advanced pancreatic adenocarcinoma. However, the effect of XRT on metastatic tumors outside the field of irradiation (abscopal effect) remains largely unknown. In the current study, we examined the effect of XRT alone and in combination with capecitabine and/or celecoxib in both irradiated and lead-shielded contralateral BxPC-3 pancreatic cancer xenografts. This chemoradiation regimen was chosen based on our molecular analysis of pancreatic adenocarcinoma. EXPERIMENTAL DESIGN: Athymic mice were injected bilaterally with BxPC-3 cells and treatment was initiated 28 days postimplant. During XRT (2 Gy for 5 consecutive days, administered on days 0 and 24), one flank was irradiated whereas the rest of the body (including the contralateral tumor) was lead shielded. Capecitabine (350 mg/kg) was administered on days 0 to 13 and 24 to 37. Celecoxib was initiated in the diet at 100 ppm (equivalent to 20 mg/kg/d p.o.) and administered throughout the study. RESULTS: In irradiated xenografts, capecitabine and XRT showed synergistic anitiumor efficacy (P=0.008), which was further improved with the addition of celecoxib (P<0.001). In contralateral shielded xenografts, abscopal effects were observed. Whereas monotherapy with XRT showed significant reduction in tumor area in irradiated xenografts, growth was promoted by 23% (P<0.001) in contralateral lead-shielded tumors in the same animals relative to untreated tumors. Interestingly, synergistic antiproliferative efficacy occurred in these contralateral tumors when capecitabine was administered (P<0.001), despite being outside the irradiated field. The addition of celecoxib further inhibited tumor growth (P<0.001). This trimodal combination most effectively stabilized disease in both shielded and irradiated tumors; however, tumor eradication was not observed. There were no significant changes in thymidine phosphorylase, dihydropyrimidine dehydrogenase, or cyclooxygenase-2 mRNA levels in irradiated or lead-shielded tumors, suggesting that efficacy cannot be predicted solely from these previously identified indicators of response. Immunohistochemistry examining the proliferation marker Ki-67 showed concordance with tumor response in both irradiated and contralateral shielded xenografts. CONCLUSIONS: These results have implications in the rational design of treatment paradigms for pancreatic cancer where metastatic disease remains the primary cause of patient morbidity and abscopal effects in tumors outside the field of irradiation may affect tumor response. PMID- 16361566 TI - The distribution of the anticancer drug Doxorubicin in relation to blood vessels in solid tumors. AB - PURPOSE: Anticancer drugs gain access to solid tumors via the circulatory system and must penetrate the tissue to kill cancer cells. Here, we study the distribution of doxorubicin in relation to blood vessels and regions of hypoxia in solid tumors of mice. EXPERIMENTAL DESIGN: The distribution of doxorubicin was quantified by immunofluorescence in relation to blood vessels (recognized by CD31) of murine 16C and EMT6 tumors and human prostate cancer PC-3 xenografts. Hypoxic regions were identified by injection of EF5. RESULTS: The concentration of doxorubicin decreases exponentially with distance from tumor blood vessels, decreasing to half its perivascular concentration at a distance of about 40 to 50 mum, The mean distance from blood vessels to regions of hypoxia is 90 to 140 microm in these tumors. Many viable tumor cells are not exposed to detectable concentrations of drug following a single injection. CONCLUSIONS: Limited distribution of doxorubicin in solid tumors is an important and neglected cause of clinical resistance that is amenable to modification. The technique described here can be adapted to studying the distribution of other drugs within solid tumors and the effect of strategies to modify their distribution. PMID- 16361567 TI - Antitumor vascular strategy for controlling experimental metastatic spread of human small-cell lung cancer cells with ZD6474 in natural killer cell-depleted severe combined immunodeficient mice. AB - BACKGROUND: Small-cell lung cancer is often characterized by rapid growth and metastatic spread. Because tumor growth and metastasis are angiogenesis dependent, there is great interest in therapeutic strategies that aim to inhibit tumor angiogenesis. METHODS: The effect of ZD6474, an orally available inhibitor of vascular endothelial growth factor receptor-2 (VEGFR-2) and epidermal growth factor tyrosine kinases, was studied in experimental multiple-organ metastasis models with human small-cell lung cancer cell lines (SBC-3 or SBC-5) in natural killer cell-depleted severe combined immunodeficient mice. RESULTS: Intravenously inoculated SBC-5 cells produced experimental metastases in the liver, lung, and bone whereas SBC-3 cells produced the metastases in the liver, systemic lymph nodes, and kidneys. Daily oral treatment with ZD6474 (50 mg/kg), started on day 14 (after the establishment of micrometastases), significantly reduced the frequency of large (>3 mm) metastatic colonies (in the liver and lymph nodes) and osteolytic bone lesions. ZD6474 treatment did not significantly reduce the frequency of small (<2-3 mm) metastatic lesions found in the lung (SBC-5) or kidney (SBC-3), consistent with an antiangiogenic mechanism of action. Immunohistochemical analysis of SBC-5 metastatic deposits in the liver showed that ZD6474 treatment inhibited VEGFR-2 activation and induced apoptosis of tumor associated endothelial cells, resulting in decreasing tumor microvessel density. ZD6474 treatment was also associated with a decrease in tumor cell proliferation and an increase in tumor cell apoptosis. The antitumor effects of ZD6474 were considered likely to be due to inhibition of VEGFR-2 tyrosine kinase because gefitinib, a small-molecule inhibitor of epidermal growth factor receptor tyrosine kinase, was inactive in these models. CONCLUSIONS: These results suggest that ZD6474 may be of potential therapeutic value in inhibiting the growth of metastatic small-cell lung cancer in humans. Phase II trials with ZD6474 are currently ongoing in a range of solid tumors. PMID- 16361568 TI - T-cell responses directed against multiple HLA-A*0201-restricted epitopes derived from Wilms' tumor 1 protein in patients with leukemia and healthy donors: identification, quantification, and characterization. AB - PURPOSE: Antigens derived from the Wilms' tumor (WT1) protein, which is overexpressed in leukemias, are attractive targets for immunotherapy. Four HLA A*0201-restricted WT1-derived epitopes have been identified: WT37, WT126, WT187, and WT235. We determined the natural immunogenecity of these antigens in patients with hematologic malignancies and healthy donor. EXPERIMENTAL DESIGN: To detect very low frequencies of WT1-specific CD8+ T cells, we used quantitative reverse transcription-PCR to measure IFN-gamma mRNA production by WT1 peptide-pulsed CD8+ T cells from 12 healthy donors, 8 patients with chronic myelogenous leukemia, 6 patients with acute myelogenous leukemia, and 8 patients with acute lymphoblastic leukemia. RESULTS: Responses were detected in 5 of 8 chronic myelogenous leukemia patients, 4 of 6 patients with acute myelogenous leukemia, and 7 of 12 healthy donors. No responses were detected in patients with acute lymphoblastic leukemia. The magnitude and extent of these CD8+ T-cell responses was greater in patients with myeloid leukemias than in healthy donors. Clonotypic analysis of WT1 specific CD8+ T cells directly ex vivo in one case showed that this naturally occurring population was oligoclonal. Using fluorescent peptide-MHC class I tetramers incorporating mutations in the alpha3 domain (D227K/T228A) that abrogate binding to the CD8 coreceptor, we were able to confirm the presence of high-avidity T-cell clones within the antigen-specific repertoire. CONCLUSION: The natural occurrence of high-avidity WT1-specific CD8+ T cells in the periphery could facilitate vaccination strategies to expand immune responses against myeloid leukemias. PMID- 16361569 TI - Targeting heat shock proteins for immunotherapy in multiple myeloma: generation of myeloma-specific CTLs using dendritic cells pulsed with tumor-derived gp96. AB - PURPOSE: To develop effective immunotherapies for patients with multiple myeloma, it is important to use novel tumor antigens. Recent studies in solid tumors show that tumor-derived heat shock proteins (Hsp) can be used as immunogen; however, no such study has yet been reported in multiple myeloma. EXPERIMENTAL DESIGN: We examined whether myeloma-derived Hsp gp96 can be used as a myeloma antigen. Specific CTL lines were obtained after repeatedly stimulating T cells with autologous, HLA-A*0201+ dendritic cells pulsed with gp96 derived from HLA-A*0201+ human myeloma cell line (HMCL) U266 or primary myeloma cells. RESULTS: These T cells lysed not only gp96-pulsed dendritic cells, U266, and other HLA-A*0201+ HMCLs IM-9 and XG1 but also effectively killed HLA-A*0201+ primary myeloma cells from patients. No killing was observed against unpulsed dendritic cells, dendritic cells pulsed with control gp96, HLA-A*0201- HMCLs, and primary myeloma cells, or HLA-A*0201+ nonmyeloma cells. Cytotoxicity was mainly MHC class I/HLA A*0201 restricted, suggesting that the CTLs recognized gp96-chaperoned peptides on HLA-A*0201 that were derived from shared myeloma antigens and that myeloma cells naturally present these peptides in the context of their surface MHC molecules. Upon antigen stimulation, these T cells secreted IFN-gamma and tumor necrosis factor-alpha, indicating that they belong to type 1 T-cell subsets. CONCLUSION: These results show that these T cells are potent CTLs that are able to effectively lyse myeloma cells but not normal blood cells and also suggest that Hsps from allogeneic tumor cells may be used as vaccines to immunize patients. PMID- 16361570 TI - AdCD40L gene therapy counteracts T regulatory cells and cures aggressive tumors in an orthotopic bladder cancer model. AB - PURPOSE: The aim of this study was to develop an immunostimulating gene therapy for the treatment of orthotopic bladder carcinoma by transferring the gene for CD40L into the tumor site. CD40L stimulation of dendritic cells induces interleukin-12 expression that drives Th1 type of immune responses with activation of cytotoxic T cells. EXPERIMENTAL DESIGN: The gene for murine CD40L was transferred into bladders of tumor-bearing mice using an adenoviral vector construct. To facilitate viral uptake, the bladders were pretreated with Clorpactin. Survival of mice as well as transgene expression and immunologic effect, such as resistance to tumor challenge and presence of T regulatory cells, were monitored. RESULTS: On viral vector instillation, CD40L expression could be detected by reverse transcription-PCR. As a sign of transgene function, interleukin-12 (IL-12) expression was significantly increased. AdCD40L gene therapy cured 60% of mice with preestablished tumors. The cured mice were completely resistant to subcutaneous challenge with MB49 tumor cells, whereas the growth of a syngeneic irrelevant tumor was unaltered. Furthermore, the mRNA expression level of the T regulatory cell transcription factor Foxp3 was evaluated both in tumor biopsies and lymph nodes. There were no differences within the tumors of the different treatment groups. However, Foxp3 mRNA levels were down-regulated in the lymph nodes of AdCD40L-treated mice. Correspondingly, T cells from AdCD40L-treated mice were not able to inhibit proliferation of naive T cells as opposed to T cells from control-treated, tumor-bearing mice. CONCLUSIONS: AdCD40L gene therapy evokes Th1 cytokine responses and counteracts T regulatory cell development and/or function. PMID- 16361571 TI - Reveromycin A inhibits osteolytic bone metastasis of small-cell lung cancer cells, SBC-5, through an antiosteoclastic activity. AB - PURPOSE: The purpose of this study was to determine therapeutic effect of a novel antibiotic, reveromycin A, against osteolytic bone metastasis of human small cell lung cancer (SBC-5) cells. RESULTS: Reveromycin A induced apoptosis specifically in osteoclasts in vitro. Although reveromycin A did not inhibit SBC-5 cell proliferation, it suppressed the expression of parathyroid hormone-related peptide. Intravenous inoculation of SBC-5 cells in natural killer cell-depleted severe combined immunodeficient mice produced experimental metastases in multiple organs, including the bone. Daily administration of reveromycin A inhibited the bone metastasis, but not visceral metastasis, in a dose-dependent manner. Histologic analyses revealed that although treatment with reveromycin A did not affect the number of proliferating tumor cells, it decreased the number of osteoclasts and increased apoptotic cells in bone lesions. CONCLUSIONS: These findings suggest that reveromycin A may inhibit osteolytic bone metastasis through suppression of osteoclast activity by directly inducing apoptosis and indirectly inhibiting tumor cell-derived parathyroid hormone-related peptide production. Therefore, reveromycin A may be a novel, potent therapeutic agent against osteolytic bone metastasis of lung cancer in humans. PMID- 16361573 TI - Increased nuclear localization of transcription factor Y-box binding protein 1 accompanied by up-regulation of P-glycoprotein in breast cancer pretreated with paclitaxel. AB - PURPOSE: The Y-box binding protein 1 (YB-1) regulates expression of P glycoprotein encoded by the MDR1 gene. There have been no previous studies regarding the involvement of YB-1 in the development of resistance to paclitaxel. The present study was done to examine how paclitaxel affects the localization and expression of YB-1 in breast cancer. EXPERIMENTAL DESIGN: We evaluated the expression and localization of YB-1 and P-glycoprotein in breast cancer tissues obtained from 27 patients before and after treatment with paclitaxel. The effect of paclitaxel on localization of cellular YB-1 was examined by using GFP-YB-1. Interaction of YB-1 with the Y-box motif of the MDR1 promoters was studied by electrophoretic mobility shift assay. The effects of paclitaxel on MDR1 promoter activity were examined by luciferase assay. RESULTS: Of 27 breast cancer tissues treated with paclitaxel, nine (33%) showed translocation of YB-1 from the cytoplasm to the nucleus together with increased expression of P-glycoprotein during the course of treatment. Twelve breast cancer tissues (44%) showed neither translocation of YB-1 nor increased expression of P-glycoprotein. Nuclear translocation of YB-1 was correlated significantly with increased expression of P glycoprotein (P=0.0037). Confocal analysis indicated that paclitaxel induced nuclear translocation of green fluorescent fused YB-1 in MCF7 cells. Furthermore, binding of YB-1 to the Y-box of MDR1 promoter was increased in response to treatment with paclitaxel. In addition, MDR1 promoter activity was significantly up-regulated by paclitaxel in MCF7 cells (P<0.001). CONCLUSIONS: The results of the present study suggested that YB-1 may be involved in the development of resistance to paclitaxel in breast cancer. PMID- 16361572 TI - Focal adhesion kinase silencing augments docetaxel-mediated apoptosis in ovarian cancer cells. AB - OBJECTIVE: Docetaxel causes cell death through induction of apoptosis; however, cell death characteristics for docetaxel have not yet been fully elucidated. We examined the role of focal adhesion kinase (FAK) cleavage in docetaxel-mediated apoptosis. METHODS: FAK degradation after treatment with docetaxel was determined in both taxane-sensitive (HeyA8 and SKOV3) and taxane-resistant (HeyA8-MDR and SKOV3-TR) ovarian cancer cell lines by Western blot analysis. Cell growth was determined with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. FAK-targeting small interfering RNA (siRNA) was used to decrease FAK expression. Apoptosis and caspase activity were determined using commercially available kits. RESULTS: SKOV3 and HeyA8 cell lines were both sensitive to docetaxel (IC50 levels, 1-6.2 nmol/L), whereas the SKOV3-TR and HeyA8-MDR cells were resistant (IC50>or=250 nmol/L for both). Docetaxel induced high rates of apoptosis in SKOV3 and HeyA8 cells (84% and 66% apoptosis, respectively) but minimal apoptosis (5-8%) in SKOV3-TR and HeyA8-MDR cells. Similarly, FAK was cleaved in SKOV3 and HeyA8 cells in response to docetaxel treatment but unchanged in the resistant cells. Caspase-3 and caspase-8 activity also increased significantly in docetaxel-treated SKOV3 and HeyA8 cells but not in the taxane resistant cells. DEVD-fmk (caspase-3 blocker) was able to block both FAK cleavage and apoptosis mediated by docetaxel in SKOV3 and HeyA8 cells. FAK siRNA transfection resulted in 70% to 90% decrease in FAK levels in all cell lines within 72 hours. FAK silencing augmented docetaxel-mediated growth inhibition (5- to 8-fold increase) and apoptosis in both of the taxane-sensitive and taxane resistant cell lines. CONCLUSIONS: Docetaxel induces FAK cleavage, mediated through activation of caspase-3, in taxane-sensitive ovarian cancer cells but not in taxane-resistant cells. The absence of FAK degradation may contribute to cell survival in taxane-resistant cells. FAK silencing promotes the in vitro efficacy of docetaxel in both taxane-sensitive and taxane-resistant cell lines and may serve as a novel therapeutic approach. PMID- 16361574 TI - Dual promoter-controlled oncolytic adenovirus CG5757 has strong tumor selectivity and significant antitumor efficacy in preclinical models. AB - PURPOSE: Transcriptionally controlled oncolytic adenovirus CG5757 is engineered with two tumor-specific promoters from E2F-1 and human telomerase reverse transcriptase genes. This virus has broad anticancer spectrum and higher specificity. The objective of the current study is to show its antitumor selectivity and therapeutic potential. EXPERIMENTAL DESIGN: The antitumor specificity of E2F-1 and human telomerase reverse transcriptase 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 cytotoxicity assays followed by a newly developed ex vivo tumor culture assay. Subsequently, in vivo antitumor efficacy and toxicology studies were carried out to assess the therapeutic potential of this oncolytic agent. RESULTS: In a broad panel of cells, E2F-1 and human telomerase reverse transcriptase promoters were activated in a tumor-selective manner. Under the control of these promoters, expression of E1a and E1b genes appears only in tumor cells. This specificity is extended to viral replication and hence the cytotoxicity in a broad range of cancer cells. Furthermore, CG5757 only replicates in cancer tissues but not in normal tissues that are derived from clinical biopsies. The safety profile was further confirmed in in vivo toxicology studies, and strong efficacy was documented in several tumor xenograft models after CG5757 was given via different routes and regimens. CONCLUSIONS: CG5757 has strong antitumor selectivity and potency. It has low toxicity and has great potential as a therapeutic agent for different types of cancers. PMID- 16361575 TI - Effect of repetitive administration of Doxorubicin-containing liposomes on plasma pharmacokinetics and drug biodistribution in a rat brain tumor model. AB - PURPOSE: The incorporation of doxorubicin in long-circulating sterically stabilized liposomes (SSL-DXR) alters the pharmacokinetics and biodistribution of doxorubicin and therefore has the potential to alter the pharmacologic properties of doxorubicin. Previously, we showed that repetitive administration of SSL-DXR alters tumor vascular permeability. EXPERIMENTAL DESIGN: Here, we investigated the effect of weekly i.v. injections of SSL-DXR on plasma pharmacokinetics and drug biodistribution in the orthotopic 9L rat brain tumor model. RESULTS AND CONCLUSIONS: The pharmacokinetics of free doxorubicin (5.67 mg/kg) did not change with repeat dosing. In contrast, drug concentrations in plasma and brain tumor increased and deposition in liver and spleen decreased after administration of the second of two weekly doses of SSL-DXR. Noncompartmental analysis and descriptive pharmacokinetic models were created to test hypotheses relating to the mechanisms responsible for alterations in SSL-DXR deposition. The analysis suggested that weekly administration of SSL-DXR significantly (P < 0.05) decreased the plasma elimination rate of SSL-DXR (34%) and decreased drug deposition in liver (2-fold) and spleen (3.5-fold). The pharmacokinetic model that best captured the observed 2.5-fold increase in tumor uptake of SSL-DXR mediated by repeat dosing was one that hypothesized that the rates of drug influx/efflux into tumor were increased by the first dose of SSL-DXR. Models that accounted only for residual drug deposited in the tissue or blood by the first weekly injection provided inferior fits to the data. Thus, the effects of repetitive dosing on SSL-DXR deposition in tumor are consistent with a treatment mediated alteration of tumor vascular permeability. PMID- 16361576 TI - Heat-induced up-regulation of NAD(P)H:quinone oxidoreductase potentiates anticancer effects of beta-lapachone. AB - PURPOSE: The purpose of the present study was to evaluate the efficacy of mild hyperthermia to potentiate the anticancer effects of beta-lapachone (3,4-dihydro 2,2-dimethyl-2H-naphthol[1,2-b]pyran-5,6-dione) by up-regulating NAD(P)H:quinone oxidoreductase (NQO1) in cancer cells. EXPERIMENTAL DESIGN: Effects of beta lapachone alone or in combination with mild heating on the clonogenic survival of FSaII fibrosarcoma cells of C3H mice and A549 human lung tumor cells in vitro was determined. Effects of heating on the NQO1 level in the cancer cells in vitro were assessed using Western blot analysis for NQO1 expression, biochemical determination of NQO1 activity, and immunofluorescence microscopy for NQO1 expression. Growth of FSaII tumors in the hind legs of C3H mice was determined after treating the host mice with i.p. injection of 45 mg/kg beta-lapachone followed by heating the tumors at 42 degrees C for 1 hour every other day for four times. RESULTS: Incubation of FSaII tumor cells and A549 tumor cells with beta-lapachone at 37 degrees C reduced clonogenic survival of the cells in dose dependent and incubation time-dependent manner. NQO1 level in the cancer cells in vitro increased within 1 hour after heating at 42 degrees C for 1 hour and remained elevated for >72 hours. The clonogenic cell death caused by beta lapachone increased in parallel with the increase in NQO1 levels in heated cells. Heating FSaII tumors in the legs of C3H mice enhanced the effect of i.p.-injected beta-lapachone in suppressing tumor growth. CONCLUSION: We observed for the first time that mild heat shock up-regulates NQO1 in tumor cells. The heat-induced up regulation of NQO1 enhanced the anticancer effects of beta-lapachone in vitro and in vivo. PMID- 16361577 TI - Enhancement of sensitivity to tumor necrosis factor alpha in non-small cell lung cancer cells with acquired resistance to gefitinib. AB - Tumor cells that have acquired resistance to gefitinib through continuous drug administration may complicate future treatment. To investigate the mechanisms of acquired resistance, we established PC-9/ZD2001, a non-small-cell lung cancer cell line resistant to gefitinib, by continuous exposure of the parental cell line PC-9 to gefitinib. After 6 months of culture in gefitinib-free conditions, PC-9/ZD2001 cells reacquired sensitivity to gefitinib and were established as a revertant cell line, PC-9/ZD2001R. PC-9/ZD2001 cells showed collateral sensitivity to several anticancer drugs (vinorelbine, paclitaxel, camptothecin, and 5-fluorouracil) and to tumor necrosis factor alpha (TNF-alpha). Compared with PC-9 cells, PC-9/ZD2001 cells were 67-fold more sensitive to TNF-alpha and PC 9/ZD2001R cells were 1.3-fold more sensitive. Therefore, collateral sensitivity to TNF-alpha was correlated with gefitinib resistance. PC-9/ZD2001 cells expressed a lower level of epidermal growth factor receptor (EGFR) than did PC-9 cells; this down-regulation was partially reversed in PC-9/ZD2001R cells. TNF alpha-induced autophosphorylation of EGFR (cross-talk signaling) was detected in all three cell lines. However, TNF-alpha-induced Akt phosphorylation and IkappaB degradation were observed much less often in PC-9/ZD2001 cells than in PC-9 cells or PC-9/ZD2001R cells. Expression of the inhibitor of apoptosis proteins c-IAP1 and c-IAP2 was induced by TNF-alpha in PC-9 and PC-9/ZD2001R cells but not in PC 9/ZD2001 cells. This weak effect of EGFR on Akt pathway might contribute to the TNF-alpha sensitivity of PC-9/ZD2001 cells. These results suggest that therapy with TNF-alpha would be effective in some cases of non-small-cell lung cancer that have acquired resistance to gefitinib. PMID- 16361578 TI - Prediction of colorectal neoplasia by quantitative methylation analysis of estrogen receptor gene in nonneoplastic epithelium from patients with ulcerative colitis. AB - PURPOSE: The incidence of colorectal neoplasia has increased among patients with longstanding and extensive ulcerative colitis (UC). Therefore, surveillance colonoscopy has been widely recommended. However, there is controversy about the impact of cancer surveillance, and ways to improve its effectiveness are being sought. The estrogen receptor (ER) gene shows age-related methylation in the colorectal epithelium and is frequently methylated in colorectal neoplasia, suggesting that ER methylation occurs early in the process of colorectal tumorigenesis. EXPERIMENTAL DESIGN: To clarify whether methylation analysis of the ER gene in nonneoplastic epithelium can help predict an increased risk for UC associated neoplasia, a total of 105 nonneoplastic colorectal epithelia from 18 patients with longstanding and extensive UC, including 8 patients with neoplasia and 10 patients without neoplasia, were analyzed. In all patients, multiple samples were taken from six regions of the colorectum. The combined bisulfite restriction analysis method was used to determine the methylation status of the ER gene. RESULTS: The mean methylation level of the ER gene was 25.4% in the nonneoplastic epithelia from UC patients with neoplasia, whereas it was only 4.0% in those without neoplasia (P<0.001). The methylation level of the ER gene in UC patients with neoplasia was significantly higher than in UC patients without neoplasia throughout the colorectum except for the cecum. In UC patients with neoplasia, the mean ER methylation level in the distal colon (36.1%) was significantly higher than in the proximal colon (14.6%; P<0.001). CONCLUSIONS: These results suggest that the analysis of ER gene methylation in nonneoplastic colorectal epithelium could have the potential to be a useful adjunct for identifying individuals with longstanding and extensive UC who are at increased risk of neoplasia and contribute to more effective cancer surveillance. PMID- 16361579 TI - In vivo molecular prediction of carbonic anhydrase IX-G250MN expression on immunotherapy outcome in renal cancer. PMID- 16361580 TI - Circulating matrix metalloproteinase-9 levels as a biomarker of disease. PMID- 16361581 TI - Shared mechanisms of ischaemic and haemorrhagic stroke: still a lot to learn. PMID- 16361582 TI - Multiple sclerosis related fatigue. PMID- 16361583 TI - "One person yawning sets off everyone else". PMID- 16361584 TI - A prospective longitudinal study of apathy in Alzheimer's disease. AB - BACKGROUND: Apathy and depression are the most frequent behavioural and psychiatric disorders in Alzheimer's disease, and may both have a negative impact on the progression of the illness. OBJECTIVES: To examine the clinical correlates of apathy in Alzheimer's disease (AD), and to determine whether apathy is a significant predictor of more rapid cognitive, functional and emotional decline. METHODS: Using a structured psychiatric evaluation, we examined a consecutive series of 354 subjects meeting clinical criteria for AD. Apathy was assessed by the Apathy Scale, and diagnosed using standardised criteria. Additional measurements included scales for depression, functional impairment, and global cognitive functions. A follow up evaluation was carried out in 247 patients (70% of the total sample) between 1 and 4 years after the baseline evaluation. RESULTS: Apathy was significantly associated with older age (p = 0.009), and a higher frequency of minor and major depression (p < 0.0001). Apathy at baseline was a significant predictor of depression at follow up (p = 0.01), and was associated with a faster cognitive (p = 0.0007) and functional decline (p = 0.006). CONCLUSIONS: Apathy in AD is a behavioural marker of a more aggressive dementia, characterised by a faster progression of cognitive, functional, and emotional impairment. PMID- 16361585 TI - Operative techniques and morbidity with subthalamic nucleus deep brain stimulation in 100 consecutive patients with advanced Parkinson's disease. AB - OBJECTIVE: Subthalamic nucleus (STN) stimulation for patients with medically refractory Parkinson disease (PD) is expanding. Reported experience has provided some indication of techniques, efficacy, and morbidity, but few centres have reported more than 50 patients. To expand this knowledge, we reviewed our experience with a large series of consecutive patients. METHODS: From March 1999 to September 2003, 191 subthalamic stimulator devices (19 unilateral) were implanted in 100 patients with PD at New York Presbyterian Hospital/Columbia University Medical Center. Sixteen patients had undergone a prior surgery for PD (pallidotomy, thalamotomy, or fetal transplant). Microelectrode guided implantations were performed using techniques similar to those described previously. Electrode implantation occurred 1-2 weeks before outpatient pulse generator implantation. RESULTS: Reductions of dyskinesias and off severity/duration were similar to prior published reports. Morbidity included: 7 device infections (3.7%), 1 cerebral infarct, 1 intracerebral haematoma, 1 subdural haematoma, 1 air embolism, 2 wound haematomas requiring drainage (1.0%), 2 skin erosions over implanted hardware (1.0%), 3 periprocedural seizures (1.6%), 6 brain electrode revisions (3.1%), postoperative confusion in 13 patients (6.8%), and 16 battery failures (8.4%). Of the 100 patients, there were no surgical deaths or permanent new neurological deficits. The average hospital stay for all 100 patients was 3.1 days. CONCLUSION: Subthalamic stimulator implantation in a large consecutive series of patients with PD produced significant clinical improvement without mortality or major neurological morbidity. Morbidity primarily involved device infections and hardware/wound revisions. PMID- 16361586 TI - Expanded motor and psychiatric phenotype in autosomal dominant Segawa syndrome due to GTP cyclohydrolase deficiency. AB - BACKGROUND: Segawa syndrome due to GTP cyclohydrolase deficiency is an autosomal dominant disorder with variable expression, that is clinically characterised by l dopa responsive, diurnally fluctuating dystonia and parkinsonian symptoms. OBJECTIVE: To delineate the neurological and psychiatric phenotype in all affected individuals of three extended families. METHODS: GTP cyclohydrolase deficiency was documented by biochemical analyses, enzymatic measurements in fibroblasts, and molecular investigations. All affected individuals were examined neurologically, and psychiatric data were systematically reviewed. RESULTS: Eighteen affected patients from three families with proven GTP cyclohydrolase deficiency were identified. Eight patients presenting at less than 20 years of age had typical motor symptoms of dystonia with diurnal variation. Five family members had late-presenting mild dopa-responsive symptoms of rigidity, frequent falls, and tendonitis. Among mutation carriers older than 20 years of age, major depressive disorder, often recurrent, and obsessive-compulsive disorder were strikingly more frequent than observed in the general population. Patients responded well to medication increasing serotonergic neurotransmission and to l dopa substitution. Sleep disorders including difficulty in sleep onset and maintenance, excessive sleepiness, and frequent disturbing nightmares were present in 55% of patients. CONCLUSION: Physicians should be aware of this expanded phenotype in affected members of families with GTP cyclohydrolase deficiency. PMID- 16361587 TI - Insulin-like growth factor I promoter polymorphism, risk of stroke, and survival after stroke: the Rotterdam study. AB - BACKGROUND AND PURPOSE: Low levels of insulin-like growth factor I (IGF-I) predispose to atherosclerosis and may therefore increase the risk of stroke. Low levels have also been found to influence the outcome of cardiovascular and cerebrovascular disease. A polymorphism in the promoter region of the IGF-I gene influences IGF-I levels. Non-carriers of the 192 bp allele have lower levels of IGF-I compared with 192 bp allele carriers. We studied the IGF-I polymorphism in relation to the risk of stroke and survival after stroke. METHODS: We studied 6808 subjects of the Rotterdam Study, who were followed for the occurrence of stroke and death after stroke. Subjects were grouped according to the 192 bp allele of IGF-I into non-carriers, heterozygotes, and homozygotes. The risk of stroke and survival after stroke was studied using Cox regression analysis, adjusting for age and sex, with homozygotes for the wildtype allele as the reference. RESULTS: Non-carriers had a relative risk of 0.8 (95% CI: 0.6 to 1.0) for the occurrence of any stroke and 0.7 (95% CI: 0.5 to 1.0) for ischaemic stroke. For non-carriers, the relative risk of death after any stroke was 1.5 (95% CI: 1.0 to 2.2). After an ischaemic stroke, this relative risk was 1.5 (95% CI: 0.9 to 2.6) and after a haemorrhagic stroke 5.2 (95% CI: 1.3 to 21.5). CONCLUSIONS: Our study suggests that IGF-I is a significant determinant of survival after stroke. PMID- 16361588 TI - Cognitive profile of subcortical ischaemic vascular disease. AB - OBJECTIVES: Subcortical ischaemic vascular disease (SIVD) is a subtype of vascular cognitive impairment characterised by extensive white matter lesions and multiple lacunar infarcts. Radiologically defined diagnostic criteria for SIVD have been introduced, but only a few studies have presented empirical data on its clinical and cognitive features. The aim of this study is to describe in detail the neuropsychological characteristics of patients with SIVD from a large well defined stroke cohort. METHODS: A sample of 323 consecutive patients with ischaemic stroke, aged 55-85 years, was investigated using neuropsychological examination and magnetic resonance imaging (MRI). Patients fulfilling the MRI criteria of SIVD (n = 85) were compared to the other stroke patients (n = 238) and to normal control subjects (n = 38). RESULTS: Cognitive performance of the SIVD group was inferior to that of the normal control group throughout all domains. As compared to the other stroke patients, the SIVD group performed significantly worse in tests measuring executive functions and delayed memory recall. Adjusting for depression had no effect on these results. Instead, after controlling for medial temporal lobe atrophy, the differences disappeared for delayed memory but remained significant for executive functions. CONCLUSION: Executive deficits are the most prominent cognitive characteristic associated with SIVD. Patients with SIVD also exhibit subtle deficits in delayed memory, which is explained in part by medial temporal lobe atrophy. Cognitive and mood changes seem to be parallel but independent processes related to SIVD. The results support the concept of SIVD as a separate clinical entity. PMID- 16361589 TI - Fatigue in multiple sclerosis: an example of cytokine mediated sickness behaviour? AB - BACKGROUND: Fatigue is a major complaint of multiple sclerosis (MS) patients. However, little is known about its pathophysiological mechanisms. Evidence from chronic fatigue syndrome and studies on sickness behaviour suggest that immune and neuroendocrine factors may play a causative role in the development of fatigue. METHODS: We compared whole blood stimulatory capacity for pro- (TNFalpha, IFNgamma) and anti-inflammatory cytokines (IL-10) as well as hypothalamo-pituitary-adrenal (HPA) axis function in 15 MS patients with marked fatigue and 15 patients without fatigue as determined by the Fatigue Severity Scale (FSS). RESULTS: Proinflammatory cytokines were significantly higher (TNFalpha: 478.9 v 228.2 pg/ml, p = 0.01; IFNgamma: 57.6 v 27.8 pg/ml; p = 0.01) in MS patients with fatigue. Furthermore, TNFalpha values significantly correlated with daytime sleepiness as measured by the Epworth Sleepiness Scale (r = 0.64, p = 0.001). Controlling for disease activity (as measured by the Cambridge Multiple Sclerosis Basic Score), disease duration, Expanded Disability Status Scale, and depression further increased the correlation of cytokine production and fatigue. HPA axis activity was not related to fatigue but was modestly correlated with cognitive impairment. CONCLUSION: Our data suggest that fatigue in MS is at least partially mediated through activation of proinflammatory cytokines. In line with earlier findings, HPA axis dysfunction seems not to be relevant in MS fatigue pathogenesis but appears to be linked to cognitive impairment. Our findings suggest that increased levels of inflammatory cytokines may be involved in MS fatigue. Investigation of cytokine profiles may increase the understanding of fatigue pathogenesis in MS. PMID- 16361590 TI - Abnormalities in normal appearing tissues in early primary progressive multiple sclerosis and their relation to disability: a tissue specific magnetisation transfer study. AB - BACKGROUND: Patients with primary progressive multiple sclerosis (PPMS) often develop severe disability despite low levels of abnormality on conventional magnetic resonance imaging (MRI). This may relate to diffuse pathological processes occurring in normal appearing brain tissue (NABT) involving both white matter (NAWM) and grey matter (NAGM). Magnetisation transfer imaging (MTI) is capable of identifying these processes and may be particularly informative when applied to patients with early PPMS. AIM: To assess the relationship between abnormalities in NABT identified by MTI and disability and other radiological data in patients with early PPMS. METHODS: We studied 43 patients within 5 years of disease onset and 43 controls. The Expanded Disability Status Scale (EDSS) and the Multiple Sclerosis Functional Composite (MSFC) were scored. Magnetisation transfer ratios (MTR) of NABT, NAWM, and NAGM were calculated and the following MTR parameters were measured: mean, peak height, peak location, and MTR value at the 25th, 50th, and 75th percentiles. Proton density, T2, T1, and gadolinium enhancing lesion loads were also calculated. RESULTS: Differences were found between patients and controls in mean, peak height, and peak location of NAWM and NAGM (p < or = 0.001). Weak to moderate correlations were found between MTR parameters and disability in both NAWM and NAGM. Strong correlations between MTR parameters and lesion loads were found, particularly in NAWM. CONCLUSION: MTR abnormalities are seen in NAWM and NAGM in early PPMS and both are associated with disability. NAWM MTR abnormalities are more closely related to conventional MRI measures than those seen in NAGM. PMID- 16361591 TI - Intense T cell depletion followed by autologous bone marrow transplantation for severe multiple sclerosis. AB - BACKGROUND: Certain stem cell transplantation procedures might slow down inflammatory pathology in multiple sclerosis (MS). AIMS: To halt disease progression in aggressive MS by a bone marrow transplantation (BMT) protocol aimed at maximum T cell suppression. METHODS: Autologous BMT was performed in 14 patients with rapid secondary progressive MS (median EDSS score at baseline, 6; median disease duration, five years). To accomplish rigorous T cell ablation, a strong conditioning protocol was chosen--cyclophosphamide, total body irradiation, and antithymocyte globulin. To minimise the possibility of reinfusing mature T cells in the graft, bone marrow, not peripheral blood, was used as the CD34+ stem cell source. RESULTS: Median follow up was 36 months (range, 7-36). Post-transplant haemopoietic recovery was successful in all patients. Early toxicity included Epstein-Barr virus related post-transplantation lymphoproliferative disorder. Longterm effects were development of antithyroid antibodies (three) and myelodysplastic syndrome (one). One patient died of progressive disease five years after transplantation. Treatment failure, defined by EDSS increase sustained for six months or more, was seen in nine patients and stabilisation or improvement in five. Other clinical parameters generally showed the same outcome. No gadolinium enhanced lesions were seen on post-treatment magnetic resonance imaging, in either cerebral or spinal cord scans. However, cerebrospinal fluid oligoclonal bands remained positive in most cases. CONCLUSIONS: This strong immunosuppressive regimen did not prevent clinical progression in patients with aggressive secondary MS. The lack of efficacy, together with some serious side effects, does not favour the use of similar rigorous T cell depleting protocols in the future. PMID- 16361592 TI - Increasing cord atrophy in early relapsing-remitting multiple sclerosis: a 3 year study. AB - OBJECTIVES: Previous studies have shown that upper cervical cord atrophy (UCCA) occurs in multiple sclerosis (MS), particularly in those disabled and with primary or secondary progressive disease. It is less clear how early it can be detected in relapsing-remitting (RR) MS, and whether early cord atrophy relates to the concurrent or future clinical course. METHODS: Twenty seven RR MS patients (median disease duration 1.7 years, in all cases <3 years from onset) were recruited along with 20 controls. They were followed for up to 3 years with a yearly assessment of UCCA and clinical function measured by the Expanded Disability Status Scale (EDSS) and MS Functional Composite Score (MSFC). Clinical and MRI correlations were investigated. Statistical models adjusted for covariates including total intracranial volume. RESULTS: Longitudinal analysis showed a significant decrease in UCCA in patients both within the patient cohort (p < 0.001) and in comparison with controls (p = 0.001). There was a significant increase in EDSS (p = 0.008) but no significant change in MSFC. The rate of UCCA loss did not correlate with clinical change or with change in brain volume. CONCLUSIONS: In summary, serial UCCA measurement detects the development of spinal cord atrophy in clinically early RR MS. PMID- 16361593 TI - Effects of sleep deprivation on cortical excitability in patients affected by juvenile myoclonic epilepsy: a combined transcranial magnetic stimulation and EEG study. AB - OBJECTIVE: To investigate the effect of sleep deprivation on corticospinal excitability in patients affected by juvenile myoclonic epilepsy (JME) using different transcranial magnetic stimulation (TMS) parameters. METHODS: Ten patients with JME and 10 normal subjects underwent partial sleep deprivation. Motor threshold (MT), motor evoked potential amplitude (MEP), and silent period (SP) were recorded from the thenar eminence (TE) muscles. Short latency intracortical inhibition (SICI) and short latency intracortical facilitation (SICF) were studied using paired magnetic stimulation. TMS was performed before and after sleep deprivation; EEG and TMS were performed simultaneously. RESULTS: In patients with JME, sleep deprivation induced a significant decrease in SICI and an increase in SICF, which was associated with increased paroxysmal activity. A significant decrease in the MT was observed. No significant changes in any TMS parameters were noted in normal subjects after sleep deprivation. The F wave was unchanged by sleep deprivation in both control subjects and in patients with JME. CONCLUSIONS: In patients with JME, sleep deprivation produces increases in corticospinal excitability in motor areas as measured by different TMS parameters. PMID- 16361594 TI - Amantadine for treatment of fatigue in Guillain-Barre syndrome: a randomised, double blind, placebo controlled, crossover trial. AB - OBJECTIVE: Fatigue is a major complaint in patients with immune mediated polyneuropathies. Despite apparently good physical recovery after Guillain-Barre syndrome (GBS), many patients remain restricted in daily and social activities, and have a decreased quality of life. In this trial, the effect of amantadine on severe fatigue related to GBS was studied. METHODS: During the pre-treatment phase, all patients were monitored for 2 weeks. Only patients with severe fatigue, defined as a mean fatigue score of > or = 5.0 on the Fatigue Severity Scale (FSS), were randomised for this double blind, placebo controlled, crossover study. Primary outcome measure was improvement of at least 1 point on the FSS. Secondary outcome measures were impact of fatigue, anxiety and depression, handicap, and quality of life. RESULTS: In total, 80 patients with GBS were randomised, of whom 74 were included for analysis. Fatigue appeared to be reduced already during the pre-treatment phase (p = 0.05), probably due to increased attention provided to the patients. No significant differences in any of the primary and secondary outcome measures were found. CONCLUSIONS: Amantadine was not superior to placebo. Because fatigue remains a serious complaint, other studies evaluating new treatment options are strongly recommended. PMID- 16361595 TI - Long term prognosis of chronic inflammatory demyelinating polyneuropathy: a five year follow up of 38 cases. AB - BACKGROUND: Little is known about long term prognosis and course after immune treatments in chronic inflammatory demyelinating polyneuropathy (CIDP). OBJECTIVE: To study long term outcomes and prognostic factors in patients with CIDP. METHODS: Clinical and electrophysiological findings, responses to immune modulating treatments, and outcomes five years after the start of treatment were reviewed in 38 CIDP patients. RESULTS: Patients were treated with corticosteroids (89%), immunoglobulin infusion (45%), or plasmapheresis (34%), and 58% received combined therapy. Five years after treatment was begun, 10 (26%) of the patients had complete remission (lasting >2 years with normal nerve conduction studies), and 23 (61%) had partial remission (able to walk) with (26%) or without (34%) immune treatments. The remaining five patients (13%) still had severe disability (unable to walk) or treatment dependent relapses. Patients with complete remission more often had subacute onset, symmetrical symptoms, good response to initial corticosteroid treatment, and nerve conduction abnormalities predominant in the distal nerve terminals. In contrast, insidious onset, asymmetrical symptoms, and electrophysiological evidence of demyelination in the intermediate nerve segments were associated with refractoriness to treatment or treatment dependent relapse. CONCLUSIONS: The long term prognosis of CIDP patients was generally favourable, but 39% of patients still required immune treatments and 13% had severe disabilities. Mode of onset, distribution of symptoms, and electrophysiological characteristics may be prognostic factors for predicting a favourable outcome. PMID- 16361596 TI - Psychosocial adjustment 17 years after severe brain injury. AB - OBJECTIVES: To examine very long term psychosocial outcome following severe brain injury in a large cohort, with the aim of evaluating Thomsen's observation that even after very serious head trauma the long term outcome in some patients is reasonably good. METHODS: The cohort consisted of 80 patients who had suffered severe brain injury evaluated at a mean time of 17 years post injury (range 10-32 years). Information regarding employment status and relationship status was obtained during clinical interview. Psychosocial outcome measures included the Supervision Rating Scale, Satisfaction with Life Scale, Hospital Anxiety and Depression Scale (HADS), Patient Competency Rating Scale, and Community Integration Questionnaire. RESULTS: Of the cohort, 72.0% lived independently, 28.7% were in full time employment, and 60.0% were married or cohabiting. The mean rating of life satisfaction was "slightly dissatisfied", but no serious emotional problems were evident from self report ratings on the HADS. Mean functional competency ratings and community integration levels were just below those reported for non-disabled patients. CONCLUSIONS: Results indicate that although long term psychosocial functioning in patients with severe head injury remains compromised, long term adjustment may be better than expected from data reported by studies assessing psychosocial outcome at earlier stages of recovery. PMID- 16361597 TI - Neurological picture. Bilateral hypertrophic olivary degeneration. PMID- 16361598 TI - Leigh syndrome caused by mutations in the flavoprotein (Fp) subunit of succinate dehydrogenase (SDHA). AB - Detailed clinical, neuroradiological, histological, biochemical, and genetic investigations were undertaken in a child suffering from Leigh syndrome. The clinical symptoms started at age five months and led to a severe progressive neurodegenerative disorder causing epilepsy, psychomotor retardation, and tetraspasticity. Biochemical measurement of skeletal muscle showed a severe decrease in mitochondrial complex II. Sequencing of SDHA revealed compound heterozygosity for a nonsense mutation in exon 4 (W119X) and a missense mutation in exon 3 (A83V), both absent in normal controls. In six additional patients- five with Leigh or Leigh-like syndrome and one with neuropathy and ataxia associated with isolated deficiency of complex II--mutations in SDHA were not detected, indicating genetic heterogeneity. PMID- 16361600 TI - Meralgia paraesthetica (Bernhardt-Roth syndrome). PMID- 16361599 TI - Early circulating levels of endothelial cell activation markers in aneurysmal subarachnoid haemorrhage: associations with cerebral ischaemic events and outcome. AB - OBJECTIVE: To investigate the relation of endothelial cell activation with delayed cerebral ischaemia (DCI) and outcome after subarachnoid haemorrhage (SAH). METHODS: Concentrations of soluble (s) intercellular adhesion molecule-1, sE-selectin, sP-selectin, ED1-fibronectin, von Willebrand Factor (vWf), and vWf propeptide were measured within three days of SAH onset. The associations with poor outcome were investigated at three months in 106 patients. In 90 patients in whom the occurrence of cerebral ischaemia could be dated accurately, two analyses were undertaken: one for all ischaemic events (n = 32), including those related to treatment, and another for spontaneous DCI (n = 11). Concentrations of markers were dichotomised at their medians. The associations of endothelial cell activation markers with outcome were expressed as odds ratios (OR) from logistic regression and those with ischaemic events as hazard ratios (HR) derived from Cox regression. RESULTS: Early vWf concentrations were associated with poor outcome (crude OR = 4.6 (95% CI, 2.0 to 10.9; adjusted OR = 3.3 (1.1 to 9.8). Early levels of vWf were also positively related to occurrence of all ischaemic events (crude HR = 2.3 (1.1 to 4.9); adjusted HR = 1.8 (0.8 to 3.9) and with occurrence of spontaneous DCI (crude HR = 3.5 (0.9 to 13.1); adjusted HR = 2.2 (0.5 to 9.8). None of the other markers showed any associations. CONCLUSIONS: Concentrations of sICAM-1, sP-selectin, sE-selectin, and ED1-fibronectin do not predict the occurrence of DCI or outcome. The positive associations of raised early vWf concentrations with ischaemic events and poor outcome after SAH may reflect a predisposition to further ischaemic injury through formation of microthrombi in the cerebral circulation. PMID- 16361601 TI - Neurological picture. Pudendal nerve compression by pelvic varices: successful treatment with transcatheter ovarian vein embolisation. PMID- 16361602 TI - Raised CSF phospho-tau concentrations in variant Creutzfeldt-Jakob disease: diagnostic and pathological implications. AB - OBJECTIVE: To investigate whether phosphorylated tau protein (tau-pT181) is increased in variant Creutzfeldt-Jakob disease (vCJD) and if the tau-pT181/tau protein ratio is useful for distinguishing between patients with and without CJD. METHODS: CSF tau protein and tau-pT181 were measured in 50 patients with sporadic CJD (sCJD), 51 patients with vCJD, 46 sCJD controls, and 37 vCJD controls, using Innotest hTau and Innotest P-Thr181, Innogenetics. RESULTS: Concentrations of CSF tau protein were increased in sCJD (5120 v 367 pg/ml in controls, p < 0.001) and vCJD (952 v 106 pg/ml, p < 0.001); tau-pT181 was also raised in sCJD (61 v 35 pg/ml in controls, p = 0.002) and vCJD (114 v 33 pg/ml, p < 0.001). Median concentrations of tau-pT181 were higher in vCJD than in sCJD (p < 0.001). The tau pT181/tau protein ratio was lower than in controls in both sCJD (12 v 128 (p < 0.001)) and vCJD (119 v 279 (p < 0.001)). Mean tau-pT181/tau protein ratio was 10 fold higher in vCJD than in sCJD. Raised CSF tau protein had the highest efficiency for distinguishing sCJD and vCJD from controls. CONCLUSIONS: CSF tau pT181 concentrations are raised in vCJD and are higher than in sCJD. Measurement of CSF tau-pT181/tau protein ratio does not improve the diagnostic efficiency of CSF tau protein alone for either vCJD or sCJD. The higher concentration of CSF tau-pT181 found in vCJD suggests that unexplained pathogenic factors influence the phosphorylation of tau protein in vCJD patients. PMID- 16361603 TI - Predictors of risk of intracerebral haemorrhage in patients with a history of TIA or minor ischaemic stroke. AB - We developed a model identifying patients with previous cerebral ischaemia at increased risk of intracerebral haemorrhage (ICH). Based on data from eight cohorts, 107 ICHs were found to have occurred among 12 648 patients. Multivariate Cox regression analysis identified the following predictors: age (> or = 60 years, hazard ratio (HR) 2.07), blood glucose level (> or = 7 mmol/l, HR 0.33), systolic blood pressure (> or = 140 mm Hg, HR 2.17), and antihypertensive drugs (HR 1.53). The highest risk quartile was associated with five times more ICHs than the lowest quartile. PMID- 16361604 TI - Arterial hypertension as risk factor for spontaneous cervical artery dissection. A case-control study. AB - Because of the presumed non-atherosclerotic pathogenesis, the potential link between spontaneous cervical artery dissection (sCAD) and common risk factors for atherosclerosis has never been investigated systematically. Therefore, this prospective, multicentre, case-control study compared the frequency of tobacco use, hypertension, diabetes mellitus, and hypercholesterolaemia among a group of consecutive patients with sCAD (n = 153), a group of patients with ischaemic stroke, not related to CAD (non-CAD), and a group of controls. As opposed to the other variables, a trend towards a significant association was seen when the prevalence of hypertension was compared among patients with sCAD and controls (26.8% v 17.0%; odds ratio (OR) 1.79; 95% confidence interval (CI), 0.98 to 3.27; p = 0.058). Hypertension was also significantly associated with the subgroup of patients with sCAD and cerebral infarction (OR, 1.94; 95% CI, 1.01 to 3.70; p = 0.045), particularly when involving the vertebral arteries (OR, 2.69; 95% CI, 1.20 to 6.04; p = 0.017). These findings might help define the spectrum of pathogenic conditions predisposing to sCAD and provide information to help investigate the combined effect of such susceptibility factors in future studies. PMID- 16361605 TI - The long term retention of levetiracetam in a large cohort of patients with epilepsy. AB - Levetiracetam (Lev) is a new antiepileptic drug with a distinct mechanism of action, shown in regulatory trials to be effective. These controlled trials do not always predict how useful a drug will be in day to day clinical practice. Retention rates can provide a better indication of efficacy and tolerability in everyday use. Patients attending a tertiary referral centre for epilepsy and who received Lev in the first 2 years of its marketing were assessed (n = 811) to determine continuation rates of treatment with this drug. At the last follow up, 65% of patients were still taking Lev, and the estimated 3 year retention rate was 58%. In total, 11% attained seizure freedom of at least 6 months. Patients taking greater numbers of concurrent antiepileptic drugs (AEDs) were more likely to discontinue Lev, and those reaching higher maximum daily dosages were less likely to discontinue Lev. The retention rate for Lev compares favourably with that of other new AEDs. PMID- 16361606 TI - Cortical laminar necrosis related to prolonged focal status epilepticus. AB - Cortical laminar necrosis (CLN) is radiologically defined as high intensity cortical lesions on T1 weighted MRI images following a gyral distribution. Histopathologically, CLN is characterised by pannecrosis of the cortex involving neurones, glial cells, and blood vessels. It has been reported to be associated with hypoxia, metabolic disturbances, drugs, and infections. We present two patients who developed CLN and permanent neurological deficits after prolonged and repeated focal status epilepticus. The possible mechanisms leading to CLN in these patients are discussed, together with the implications of prompt and aggressive treatment in similar cases. PMID- 16361607 TI - Hemifacial motor and crying seizures of temporal lobe onset: case report and review of electro-clinical localisation. AB - OBJECTIVE: To report a case of temporal lobe epilepsy with clinical presentation of paroxysmal episodes of "tightness" over the right hemiface, and ictal crying, and review electroclinical localisation of this phenomenon. METHODS: Clinical semiology, neurophysiological localising tests, and epilepsy surgery outcome are reported in a subject presenting with paroxysmal right hemifacial movements and ictal crying. Pertinent past reports of somato-motor signs and ictal crying in temporal lobe epilepsy are reviewed and the findings correlated with proposed human facial cortical representation. RESULTS: Simple partial seizures caused by temporal lobe epilepsy presented with right sided tonic facial movements and ictal crying. Intracranial EEG monitoring documented a left medial temporal onset of seizures that remained asymptomatic until they propagated to the left cingulate region. Anterior temporal lobectomy with resection of the amygdala and anterior hippocampus resulted in cessation of seizures. CONCLUSIONS: This is a rare example of epileptic seizures of medial temporal onset presenting with isolated somato-motor manifestations and ictal crying. Anatomical-electrical clinical correlations with cortical regions controlling facial movements were highly suggestive that this case represents secondary activation of "emotional" motor cortex M3 and M4 (rostral and caudal cingulate motor cortex), giving rise to focal hemifacial movements and ictal crying. PMID- 16361608 TI - Anti-Ma2 associated paraneoplastic neurological syndrome presenting as encephalitis and progressive muscular atrophy. AB - A 36 year old man with a history of testicular germ cell tumour presented six months after bilateral orchidectomy with progressive amnesia, irritability, vertical gaze palsy, and generalised seizures. Eight months after initial onset of symptoms, he demonstrated a head drop with muscular atrophy of the upper limbs, shoulder girdle, and posterior neck. He reported no sensory disturbances and his sensory examination was normal. The overall clinical presentation was consistent with motor neurone disease. Cerebrospinal fluid analysis revealed mild pleocytosis and increased protein concentration. Serum and cerebrospinal fluid were positive for the anti-Ma2 antibody by western blot analysis and immunostaining. Abnormal high signal in the grey matter was noted in the cervical spinal cord and brain by T2 weighted magnetic resonance imaging (MRI). The patient was treated with corticosteroids, intravenous immunoglobulin, and antiepileptic medication. The patient improved clinically and symptom progression ceased after initiation of treatment. There was complete resolution of the abnormal brain MRI lesions; however, the cervical spinal cord MRI lesion and muscular atrophy remained unchanged. It is suggested that the anti-Ma2 antibody is involved not only in encephalitis, but may also play a role in the cervical spinal cord lesions resulting in a motor neurone disease-like presentation. PMID- 16361609 TI - Predicting response to treatment in chronic inflammatory demyelinating polyradiculoneuropathy. AB - OBJECTIVE: To discover whether Inflammatory Neuropathy Cause and Treatment Group (INCAT) electrophysiological criteria for demyelinating neuropathy predict response to immunotherapy in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). METHODS: This was a retrospective case note study of patients who had attended Guy's Hospital Peripheral Nerve Clinic between January 2001 and March 2004, been diagnosed as having CIDP, and given treatment with corticosteroids, intravenous immunoglobulin (IVIg), or plasma exchange (PE). Patients' nerve conduction studies (NCS) were reviewed for evidence of demyelination and whether the abnormalities fulfilled modified INCAT electrophysiological criteria. Patients whose NCS fulfilled the criteria were assigned to the neurophysiologically definite CIDP group, while those that did not were labelled as neurophysiologically probable CIDP. Responses to any of the three immunotherapy agents were compared between the two groups. RESULTS: Out of 50 patients, 27 (54%) were classified as neurophysiologically definite and 23 (46%) as neurophysiologically probable CIDP patients. Twenty (74%) neurophysiologically definite and 17 (73.9%) neurophysiologically probable CIDP patients responded to treatment. CONCLUSIONS: INCAT electrophysiological criteria did not predict a higher rate of response to immunotherapy. Neurophysiologically probable CIDP patients should be given a trial of immunotherapy. PMID- 16361610 TI - Botulinum toxin A during pregnancy: a survey of treating physicians. AB - Botulinum toxin A (btxA) is widely used for cosmetic purposes, headaches, dystonia, spasticity, pain and other on and off label uses. Despite the widespread use of btxA in women of childbearing potential, there are few data on the effects of this drug on pregnant women and the fetus. The goal of this study was to survey physicians who use btxA, to determine their experience with pregnant women. We surveyed 900 physicians who used commercially available btxA. The questionnaire asked treating physicians if they had knowingly or unknowingly injected pregnant women and what was the outcome of each pregnancy. In total, 396 physicians (44%) returned questionnaires, of whom only 12 physicians reported injecting pregnant women with btxA. Sixteen pregnant women were injected, mostly in the first trimester, and only one patient, who had prior spontaneous abortions, suffered a miscarriage. Another woman had a therapeutic abortion. All other pregnancies went to term and there were no fetal malformations. Based on this limited survey of treating physicians in the USA, btxA appears to be relatively safe for both expectant mother and fetus. We need further data, however, and we would recommend that physicians and patients carefully consider the risks and benefits before using btxA in pregnant women. PMID- 16361611 TI - Frontal white matter anisotropy and symptom severity of late-life depression: a magnetic resonance diffusion tensor imaging study. AB - OBJECTIVES: To investigate the disruption of neural circuits in the frontal lobes and limbic structures in late-life depressed patients compared with healthy controls, and to examine the correlation between the degree of microstructural abnormalities of white matter and clinical symptom severity in late-life depression. METHODS: Thirteen patients with late-life depression and matched control subjects underwent diffusion tensor imaging. Fractional anisotropy (FA), an index of the integrity of white matter tracts, was determined in the white matter of frontal, temporal, and occipital brain regions and the corpus callosum. RESULTS: A significant reduction was found in white matter FA values of widespread regions of the frontal and temporal lobes of depressed patients. Also, there was some evidence suggesting that white matter FA values of the inferior frontal brain region are inversely related to severity of depression. CONCLUSIONS: These results suggest the possible loss of integrity within frontal and temporal white matter fibre tracts and implicate the orbitofrontal circuit in symptom severity in late-life depression. PMID- 16361612 TI - Paraneoplastic cerebellar degeneration in olfactory neuroepithelioma. PMID- 16361613 TI - The collagen 1A2 polymorphism rs42524, which is associated with intracranial aneurysms, shows no association with spontaneous cervical artery dissection (sCAD). PMID- 16361615 TI - Neoadjuvant chemotherapy for metastatic colon cancer: a cautionary note. PMID- 16361616 TI - Phase III trial of paclitaxel plus carboplatin with or without tirapazamine in advanced non-small-cell lung cancer: Southwest Oncology Group Trial S0003. AB - PURPOSE: Tumor hypoxia confers chemotherapy resistance. Tirapazamine is a cytotoxin that selectively targets hypoxic cells. We conducted a phase III clinical trial to determine whether the addition of tirapazamine to paclitaxel and carboplatin offered a survival advantage when used in the treatment of patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Of 396 patients registered, 367 eligible patients were randomly assigned to either arm 1 (n = 181), which consisted of treatment every 21 days with paclitaxel 225 mg/m2/3 h, carboplatin (area under the curve = 6), and tirapazamine 260 mg/m2 in cycle 1 (which was escalated, if tolerable, to 330 mg/m(2) in cycle 2), or arm 2 (n = 186), which consisted of paclitaxel and carboplatin as in arm 1 with no tirapazamine. RESULTS: Patient characteristics were similar between the two arms. There were no statistically significant differences in response rates, progression-free survival, or overall survival. Patients on arm 1 had significantly (P < .05) more abdominal cramps, fatigue, transient hearing loss, febrile neutropenia, hypotension, myalgias, and skin rash and were removed from treatment more often as a result of toxicity than patients in arm 2 (26% v 13%, respectively; P = .003). More than 40% of patients did not have the tirapazamine dose escalated, primarily because of toxicity. The trial was closed early after an interim analysis demonstrated that the projected 37.5% improvement in survival (8 v 11 months median survival) in arm 1 was unachievable (P = .003). CONCLUSION: The addition of tirapazamine to paclitaxel and carboplatin does not result in improved survival in advanced NSCLC compared with paclitaxel and carboplatin alone but substantially increases toxicity. PMID- 16361617 TI - 14-3-3sigma methylation in pretreatment serum circulating DNA of cisplatin-plus gemcitabine-treated advanced non-small-cell lung cancer patients predicts survival: The Spanish Lung Cancer Group. AB - PURPOSE: Survival in patients with advanced non-small-cell lung cancer (NSCLC) who are treated with platinum-based chemotherapy is rather variable. Methylation dependent transcriptional silencing of 14-3-3sigma, a major G2-M checkpoint control gene, could be a predictor of longer survival. PATIENTS AND METHODS: A sensitive methylation-specific polymerase chain reaction assay was used to evaluate 14-3-3sigma methylation status in pretreatment serum DNA obtained from 115 cisplatin-plus-gemcitabine-treated advanced NSCLC patients. RESULTS: 14-3 3sigma methylation was observed in all histologic types of 39 patients (34%). After a median follow-up of 9.8 months, median survival was significantly longer in the methylation-positive group (15.1 v 9.8 months; P = .004). Median time to progression was 8 months in the methylation-positive group and 6.3 months in the methylation-negative group (log-rank test, P = .027). A multivariate Cox regression model identified only 14-3-3sigma methylation status and Eastern Cooperative Oncology Group performance status as independent prognostic factors for survival. In an exploratory analysis, median survival for 22 methylation positive responders has not been reached, whereas survival was 11.3 months for 29 methylation-negative responders (P = .001). CONCLUSION Methylation of 14-3-3sigma is a new independent prognostic factor for survival in NSCLC patients receiving platinum-based chemotherapy. It can be reliably and conveniently detected in the serum, thus obviating the need for tumor tissue analysis. PMID- 16361618 TI - Should elderly non-small-cell lung cancer patients be offered elderly-specific trials? Results of a pooled analysis from the North Central Cancer Treatment Group. AB - PURPOSE: To answer the question, "should elderly non-small-cell lung cancer patients be offered elderly-specific trials?" PATIENTS AND METHODS: The North Central Cancer Treatment Group (NCCTG) performed a pooled analysis of elderly patients who participated in elderly-specific trials (required age > or = 65 years) and age-unspecified trials (required age > or = 18 years). Between 1998 and 2000, all NCCTG non-small-cell lung cancer (NSCLC) patients with incurable cancer, age > or = 65 years, and receiving first-line chemotherapy were included. A total of 118 elderly patients participated in elderly-specific trials, and 108, in age-unspecified trials. Demographics and outcomes were compared based on trial type. RESULTS: The median age of elderly patients in elderly-specific trials was greater: median (range): 73 years (65 to 87) and 70 years (65 to 85), respectively (P < .001), as was the percentage older than 80 years: 17% and 3%, respectively (P = .0008). Median survival times were 232 and 302 days, respectively (P = .08). After adjustment for baseline age, Eastern Cooperative Oncology Group performance score, cancer stage, and body mass index, this survival difference was not statistically significant (hazard ratio = 1.25; P = .16). Grade 3 or worse nonhematologic adverse event rates were greater in age unspecified trials (81% v 57%, respectively; P < .001), as were grade 3 or worse hematologic events (68% v 10%, respectively; P < .001). CONCLUSION: Elderly patients in NSCLC elderly-specific trials suffered lower rates of severe adverse events with no statistically significant differences in survival. It seems that elderly-specific trials are providing quality care and helping to define optimal cancer therapy in the elderly, particularly among the "oldest of the old." PMID- 16361619 TI - Risk factors and therapy for isolated central nervous system relapse of pediatric acute myeloid leukemia. AB - PURPOSE: CNS relapse of pediatric acute myeloid leukemia (AML) is an infrequent occurrence. This review examines the risk factors and therapy used for patients with an isolated CNS relapse. PATIENTS AND METHODS: Records of 886 patients with de novo AML were reviewed, and patients who entered remission at the end of one course of therapy and developed an isolated CNS relapse as their first event were analyzed (n = 690). RESULTS: Thirty-three patients developed an isolated CNS relapse. Factors at diagnosis significantly associated with an isolated CNS relapse, compared with no CNS relapse, included age 0 to 2 years (70% v 27%, respectively; P < .001), enlarged liver (79% v 39%, respectively; P < .001) or spleen (79% v 39%, respectively; P < .001) at diagnosis, CNS disease at diagnosis (33% v 9%, respectively; P < .001), median WBC count (79.2 v 19.3 x 10(3) microL, respectively; P < .001), French-American-British M5 morphology (45% v 15%, respectively; P < .001), and chromosome 11 abnormalities (44% v 18%, respectively; P = .022). Treatment of the isolated CNS relapse varied from local therapy with intrathecal chemotherapy and/or radiation therapy to systemic therapy with chemotherapy with or without bone marrow transplantation. Survival rate in the patients treated with local therapy was only 31.5% compared with 21.4% in patients treated with systemic therapy. The 8-year overall survival for patients after an isolated CNS relapse was similar to patients after a bone marrow relapse (26% +/- 16% v 21% +/- 5%, respectively). CONCLUSION: Significant predictors for isolated CNS relapse were identified. This study demonstrated that there may be no benefit to systemic therapy versus CNS-directed therapy in outcome. The data support CNS-directed therapy to treat isolated CNS relapse. PMID- 16361620 TI - Phase I clinical and pharmacokinetic study of flavopiridol in children with refractory solid tumors: a Children's Oncology Group Study. AB - PURPOSE: To determine the dose-limiting toxicities, maximum-tolerated dose, and pharmacokinetics of the cyclin-dependent kinase inhibitor flavopiridol (NSC 649890) when administered as a 1-hour infusion over 3 consecutive days to children with recurrent or refractory solid tumors. PATIENTS AND METHODS: Flavopiridol was administered as a 1-hour intravenous infusion daily for 3 consecutive days every 21 days, or when hematologic toxicity or any grade 2 or greater nonhematologic toxicity resolved. The starting dose was 37.5 mg/m2/d. Dose escalation was in cohorts of three patients in a standard fashion until dose limiting toxicity and the maximum-tolerated dose were determined. Flavopiridol levels were measured on days 1, 2, and 3. RESULTS: Twenty-five children received flavopiridol at doses of 37.5 to 80 mg/m2/day over 3 consecutive days. The maximum-tolerated dose was 62.5 mg/m2/d. The primary dose-limiting toxicities were neutropenia and diarrhea. No antitumor effect was observed in this population. Mean peak plasma concentrations of 3.71 and 9.11 micromol/L were achieved at the end of the 1-hour infusion, following dose escalation from 37.5 mg/m2 to 80 mg/m2, respectively. The median flavopiridol plasma clearance was 8.0 L/h/m2 (range, 2.6 to 17.1 L/h/m2). CONCLUSION: The maximum-tolerated dose of flavopiridol in children, and the recommended phase II dose for pediatric studies, was 62.5 mg/m2/day when administered as a 1-hour infusion for 3 consecutive days. Dose-limiting toxicities of neutropenia and diarrhea were similar to those in adult studies. PMID- 16361621 TI - Health insurance coverage in survivors of childhood cancer: the Childhood Cancer Survivor Study. AB - PURPOSE: To examine the prevalence and predictors of health insurance coverage and the difficulties obtaining coverage in a large cohort of childhood cancer survivors. PATIENTS AND METHODS: This study included 12,358 5-year survivors of childhood cancer and 3,553 sibling controls participating in the Childhood Cancer Survivor Study. Data were collected by surveys distributed in 1994 (baseline) and 2000 (follow-up). RESULTS: At baseline, 83.9% of adult survivors, compared with 88.3% of siblings, had health insurance coverage (P < .01); 6 years later, small but significant survivor-sibling differences remained (88% v 91%; P < .01). Twenty-nine percent of survivors reported having had difficulties obtaining coverage, compared with only 3% of siblings (P < .01). In multivariate analysis of survivors 18 years of age or older, factors associated with being uninsured included younger age at diagnosis (diagnosis age of 0 to 4 years; odds ratio [OR] = 1.7; 95% CI, 1.3 to 2.2), male sex (OR = 1.3; 95% CI, 1.2 to 1.5), age at baseline survey (age 22 to 24 years; OR = 1.6; 95% CI, 1.2 to 2.1), lower level of attained education (less than high school, OR = 2.6, 95% CI, 2.1 to 3.3; high school graduate, OR = 2.1, 95% CI, 1.8 to 2.5), income less than 20,000 dollars (OR = 5.6, 95% CI, 4.5 to 7.1), marital status (widowed/divorced/separated; OR = 1.3; 95% CI, 1.1 to 1.6), smoking status (current smoker, OR = 2.0, 95% CI, 1.7 to 2.3; former smoker, OR = 1.4, 95% CI, 1.2 to 1.8), and treatment that included cranial radiation (OR = 1.3, 95% CI, 1.0 to 1.6). CONCLUSION: Compared with siblings, adult survivors of childhood cancer had significantly lower rates of health insurance coverage and more difficulties obtaining coverage. Since lack of coverage likely has serious health and financial implications for this at-risk population, any disparity in availability and quality of coverage is of great concern. PMID- 16361622 TI - Docetaxel plus gemcitabine or docetaxel plus cisplatin in advanced pancreatic carcinoma: randomized phase II study 40984 of the European Organisation for Research and Treatment of Cancer Gastrointestinal Group. AB - PURPOSE: To define the efficacy and toxicity of docetaxel plus gemcitabine or docetaxel plus cisplatin for advanced pancreatic carcinoma. PATIENTS AND METHODS: Chemotherapy-naive patients with measurable disease and WHO performance status less than 2 were randomly assigned to receive 21-day cycles of gemcitabine 800 mg/m2 on days 1 and 8 plus docetaxel 85 mg/m2 on day 8 (arm A) or docetaxel 75 mg/m2 on day 1 plus cisplatin 75 mg/m2 on day 1 (arm B). Primary end points were tumor response and rate of febrile neutropenia grade. RESULTS: Of 96 randomly assigned patients (49 patients in arm A and 47 patients in arm B), 70 patients were analyzed for response (36 in arm A and 34 in arm B) and 89 patients were analyzed for safety (45 in arm A and 44 in arm B). Confirmed responses were observed in 19.4% (95% CI, 8.2% to 36.0%) of patients in arm A and 23.5% (95% CI, 10.7% to 41.2%) in arm B. In arm A, the median progression-free survival (PFS) was 3.9 months (95% CI, 3.0 to 4.7 months), median survival was 7.4 months (95% CI, 5.6 to 11.0 months), and 1-year survival was 30%. In arm B, the median PFS was 2.8 months (95% CI, 2.6 to 4.6 months), median survival was 7.1 months (95% CI, 4.8 to 8.7 months), and 1-year survival was 16%. Febrile neutropenia occurred in 9% and 16% of patients in arms A and B, respectively. CONCLUSION: Both regimens are well tolerated and show activity in advanced pancreatic carcinoma. The safety profile and survival analyses favor docetaxel plus gemcitabine for further evaluation. PMID- 16361623 TI - Low rectal cancer: a call for a change of approach in abdominoperineal resection. AB - PURPOSE: Despite the major improvements that have been made due to total mesorectal excision (TME), low rectal cancer still remains a challenge. METHODS: By investigating a prospective randomized rectal cancer trial in which surgeons had undergone training in TME the factors responsible for the poor outcome were determined and a new method for assessing the quality of surgery was tested. RESULTS: Survival differed greatly between abdominoperineal resection (APR) and anterior resection (AR; 38.5% v 57.6%, P = .008). Low rectal carcinomas have a higher frequency of circumferential margin involvement (26.5% v 12.6%, P < .001). More positive margins were present in the patients operated with APR (30.4%) compared to AR (10.7%, P = .002). Furthermore, more perforations were present in these specimens (13.7% v 2.5%, P < .001). The plane of resection lies within the sphincteric muscle, the submucosa or lumen in more than 1/3 of the APR cases, and in the remainder lay on the sphincteric muscles. CONCLUSION: We systematically described and investigated the pathologic properties of low rectal cancer in general, and APR in particular, in a prospective randomized trial including surgeons who had been trained in TME. The poor prognosis of the patients with an APR is ascribed to the resection plane of the operation leading to a high frequency of margin involvement by tumor and perforation with this current surgical technique. The clinical results of this operation could be greatly improved by adopting different surgical techniques and possibly greater use of radiochemotherapy. PMID- 16361624 TI - Randomized phase II trial of the clinical and biological effects of two dose levels of gefitinib in patients with recurrent colorectal adenocarcinoma. AB - PURPOSE: The clinical objective of this trial was to evaluate gefitinib in patients with metastatic colorectal cancer that had progressed despite prior treatment. Serial tumor biopsies were performed when possible and analyzed for activation of the epidermal growth factor receptor (EGFR) signaling pathway. Serial serum samples were measured for amphiregulin and transforming growth factor-alpha (TGFalpha). PATIENTS AND METHODS: One hundred fifteen patients were randomly assigned to receive gefitinib 250 or 500 mg orally once a day. One hundred ten patients were assessable for clinical efficacy. Biologic evaluation was performed on paired tumor samples from 28 patients and correlated with clinical outcome. RESULTS: Median progression-free survival was 1.9 months (95% CI, 1.8 to 2.1 months) and 4-month progression-free survival rate was 13% +/- 5%. One patient achieved a radiographic partial response (RR = 1%; 95% CI, 0.01% to 5%). Median survival was 6.3 months (95% CI, 5.1 to 8.2 months). The most common adverse events were skin rash, diarrhea, and fatigue. In the biopsy cohort, expression of total or activated EGFR, activated Akt, activated MAP-kinase, or Ki67 did not decrease following 1 week of gefitinib. However, a trend toward decreased post-treatment levels of activated Akt and Ki67 was observed in patients with a PFS higher than the median, although these did not reach the .05 level of significance. CONCLUSION: Gefitinib is inactive as a single agent in patients with previously treated colorectal cancer. In tumor samples, gefitinib did not inhibit activation of its proximal target, EGFR. Trends were observed for inhibition of downstream regulators of cellular survival and proliferation in patients achieving longer progression-free survival. PMID- 16361625 TI - Dealing with a deluge of data: an assessment of adverse event data on North Central Cancer Treatment Group trials. AB - PURPOSE: Adverse events (AEs) are monitored in clinical trials for patient safety, to satisfy reporting requirements, and develop safety profiles. Recently, much attention has been placed on the reporting of serious AEs (SAEs) that are either life threatening or lethal in clinical trials. However, SAEs comprise a small subset of all AE data collected for trials; the majority of AE data collected are routine AEs (RAEs) regarding non-life-threatening events. We assessed the utility of the RAE data collected, relative to the volume. PATIENTS AND METHODS: We surveyed the RAE data from 26 North Central Cancer Treatment Group coordinated trials. RESULTS: A total of 8,318 (11%) of 75,598 of RAEs required queries. Of these, 86% were protocol-required RAEs, 83% of RAEs required per protocol were within normal limits (eg, platelets) or not present, and 61% of extra AEs were mild. One fifth of RAEs were considered unlikely to be related or unrelated to treatment. Overall, 3% of events were severe, life threatening, or caused death. Only 1% of RAE data reported required expedited reporting (eg, via Adverse Event Expedited Reporting System). Results indicate that 72% of RAEs would be eliminated if only the maximum severity per patient and type were required. These results were validated in a large phase III trial. CONCLUSION: The majority of RAEs identified, transcribed, and entered are not clinically important. Our data suggest that reducing the number of AEs monitored will affect substantially neither overall patient safety nor compromise evaluation of regimens undergoing testing. We present several considerations for such a reduction in data collection, as well as a policy that we have used to address the deluge of RAE data. PMID- 16361626 TI - Evaluation of factors affecting awareness of and willingness to participate in cancer clinical trials. AB - PURPOSE: Annually, only 3% of patients participate in cancer clinical trials (CCTs). Barriers to accrual include lack of CCT awareness and uncertain third party payer coverage. In January 2002, a California law (Senate Bill 37 [SB37]) required all third-party payers to reimburse patient care costs related to CCTs. We evaluated the level of awareness of patients and/or their family members/friends regarding CCTs and SB37. METHODS: We used both a written survey for patients and/or their family members and friends seen in oncology clinics, and a verbal telephone version for Cancer Information Service callers. We tested for correlations between CCT awareness and SB37 knowledge, and willingness to participate in CCTs. RESULTS: Of 1,188 respondents, 59% were aware of CCTs, 19% knew of SB37, and 36% were very likely to consider a CCT. There were significant positive correlations between CCT awareness and willingness to participate (P < .001, Spearman), and between SB37 knowledge and willingness to participate (P = .001, Pearson chi2). Reduced awareness was seen in respondents who were either black or African American (odds ratio [OR], 0.44; P = .004), Hispanic (OR, 0.56; P = .03), had an annual income less than 25,000 dollars (OR, 0.38; P < .001), or had less than a college degree (OR, 0.12 to 0.53; P < .001 to .013). Reduced willingness to participate in CCTs was seen in black or African American participants (OR, 0.38; P < .001), Asians (OR, 0.44; P < .006), or respondents aged 18 to 24 years (OR, 0.35; P = .002). CONCLUSION: These results support the hypothesis that improving CCT awareness and SB37 knowledge especially among lower income, less educated, and minority patients, may potentially overcome barriers to participation and subsequently increase accrual in California. PMID- 16361627 TI - Etoposide and cisplatin chemotherapy for metastatic good-risk germ cell tumors. AB - PURPOSE: To assess response, overall survival, and relapse-free survival of patients with good-risk metastatic germ cell tumor (GCT) by International Germ Cell Consensus Classification Group (IGCCCG) criteria treated with four cycles of etoposide and cisplatin (EP). PATIENTS AND METHODS: Two hundred eighty-nine patients with IGCCCG good-risk GCT were treated with four cycles of EP. EP consisted of four cycles of etoposide 100 mg/m2 and cisplatin 20 mg/m2 on days 1 to 5 every 21 days. RESULTS: Two hundred eighty-two of 289 patients (98%) achieved a complete response; 269 (93%) responded to chemotherapy alone and 13 (5%) responded to chemotherapy plus surgical resection of viable disease (GCT other than mature teratoma). Seventeen (6%) experienced relapse, and nine (3%) died as a result of disease at a median follow-up of 7.7 years (range, 0.4 to 21.1 years). Sixty-two of 204 patients (30%) with nonseminoma had findings of teratoma or viable GCT at postchemotherapy surgery. CONCLUSION: Four cycles of EP is a highly effective therapy for patients with good-risk GCT, with a high cure rate, low relapse rate, and little evidence of late relapse. Postchemotherapy surgery resection of residual disease remains an important aspect of treatment for these patients. Four cycles of EP is acceptable as a standard regimen for the treatment of good-risk metastatic GCT, and serves as an alternative to three cycles of bleomycin and etoposide before cisplatin. PMID- 16361628 TI - Dietary patterns and survival after breast cancer diagnosis. AB - PURPOSE: There is little prior study of major dietary patterns and breast cancer survival. METHODS: Patients included 2,619 Nurses' Health Study participants who were diagnosed with invasive breast cancer between 1982 and 1998 and completed a dietary questionnaire more than 1 year after diagnosis. Participants were followed through 2002 (median = 9 years). During follow-up, 414 patients died of any cause, 242 patients died of breast cancer, and 172 patients died from causes other than breast cancer. Women with in situ or metastatic disease at diagnosis were excluded. We used Cox proportional hazards models to evaluate prospective associations of prudent and Western dietary patterns assessed both before and after diagnosis with time to event after diagnosis. RESULTS: In multivariate adjusted analyses assessed after diagnosis, the Western and prudent dietary patterns were unrelated to all-cause or breast cancer mortality. However, compared with women with the lowest intake of the prudent dietary pattern, the relative risks (and 95% CIs) of death from causes other than breast cancer were 0.85 (95% CI, 0.53 to 1.35), 0.74 (95% CI, 0.45 to 1.21), 0.70 (95% CI, 0.42 to 1.17), and 0.54 (95% CI, 0.31 to 0.95; P = .03, from lowest to highest quintile of intake). In contrast, the Western dietary pattern was positively associated with this outcome (P = .04). Results for the assessment of dietary patterns before diagnosis were similar, except the prudent dietary pattern was unrelated to mortality. CONCLUSION: A higher intake of the prudent pattern and a lower intake of the Western pattern may protect against mortality from causes unrelated to breast cancer. PMID- 16361629 TI - Outcome after pathologic complete eradication of cytologically proven breast cancer axillary node metastases following primary chemotherapy. AB - PURPOSE: Pathologic complete remission (pCR) of primary breast tumors after primary chemotherapy (PCT) is associated with higher relapse-free survival (RFS) and overall survival (OS) rates. The purpose of this study was to determine long term outcome in patients achieving pCR of cytologically proven axillary lymph node (ALN) metastases. METHODS: Patients with cytologically documented ALN metastases were treated in five prospective PCT trials. After surgery, patients were subdivided into those with and without residual ALN carcinoma. Survival was calculated by the Kaplan-Meier method. RESULTS: Of 925 patients treated, 403 patients had cytologically confirmed ALN metastases. Eighty-nine patients (22%) achieved ALN pCR after PCT. Compared with the group without ALN pCR, 5-year OS and RFS were improved in patients achieving ALN pCR (93% [95% CI, 87.5 to 98.5] and 87% [95% CI, 79.7 to 94.3] v 72% [95% CI, 66.5 to 77.5] and 60% [95% CI, 54.1 to 65.9], respectively; P < .0001). Residual primary tumor did not affect outcome of those with ALN pCR. Combination anthracycline/taxane-based PCT resulted in significantly more ALN pCRs, although outcome after ALN pCR was not improved by taxanes. We constructed a nomogram demonstrating that patients who do not benefit from neoadjuvant anthracyclines are unlikely to benefit from subsequent taxanes. CONCLUSION: ALN pCR is associated with an excellent prognosis, even with a residual primary tumor, pointing to biologic differences between primary and metastatic cells. ALN pCR represents an early surrogate marker of long-term outcome. Response to initial PCT has important potential as a guide to subsequent therapy. PMID- 16361630 TI - Pharmacogenetics of tamoxifen biotransformation is associated with clinical outcomes of efficacy and hot flashes. AB - PURPOSE: Polymorphisms in tamoxifen metabolizing genes affect the plasma concentration of tamoxifen metabolites, but their effect on clinical outcome is unknown. METHODS: We determined cytochrome P450 (CYP)2D6 (*4 and *6) and CYP3A5 (*3) genotype from paraffin-embedded tumor samples and buccal cells (living patients) in tamoxifen-treated women enrolled onto a North Central Cancer Treatment Group adjuvant breast cancer trial. The relationship between genotype and disease outcome was determined using the log-rank test and Cox proportional hazards modeling. RESULTS: Paraffin blocks were obtained from 223 of 256 eligible patients, and buccal cells were obtained from 17 living women. CYP2D6 (*4 and *6) and CYP3A5 (*3) genotypes were determined from 190, 194, and 205 patient samples and in 17 living women. The concordance rate between buccal and tumor genotype was 100%. Women with the CYP2D6 *4/*4 genotype had worse relapse-free time (RF time; P = .023) and disease-free survival (DFS; P = .012), but not overall survival (P = .169) and did not experience moderate to severe hot flashes relative to women heterozygous or homozygous for the wild-type allele. In the multivariate analysis, women with the CYP2D6 *4/*4 genotype still tended to have worse RFS (hazard ratio [HR], 1.85; P = .176) and DFS (HR, 1.86; P = .089). The CYP3A5*3 variant was not associated with any of these clinical outcomes. CONCLUSION: In tamoxifen-treated patients, women with the CYP2D6 *4/*4 genotype tend to have a higher risk of disease relapse and a lower incidence of hot flashes, which is consistent with our previous observation that CYP2D6 is responsible for the metabolic activation of tamoxifen to endoxifen. PMID- 16361631 TI - Individualized survival curves improve satisfaction with cancer risk management decisions in women with BRCA1/2 mutations. AB - PURPOSE: Women with BRCA1/2 mutations are faced with complex decisions about breast and ovarian cancer risk management. This study was conducted to determine the effect of a tailored decision support system (DSS) that provides individualized survival and cancer incidence curves specific to expected outcomes of alternative management strategies. PATIENTS AND METHODS: This was a double blind, randomized controlled trial of 32 women with BRCA1/2 mutations. Primary outcome measures were decision satisfaction, cancer anxiety, perceptions of cancer risk given alternative management strategies, and management decisions. RESULTS: Twenty-seven women completed a 6-week follow-up. Women in the intervention arm (n = 13) reported significantly higher decision satisfaction at follow-up than women in the control arm (n = 14; adjusted mean difference, 9.7; P < .0005). The effect of the DSS was greater among women with low cancer anxiety at baseline than women with high cancer anxiety at baseline (P = .01 for interaction). However, the DSS did not significantly alter cancer anxiety at follow-up, perceptions of cancer risk given alternative management strategies, or management decisions. CONCLUSION: The presentation of individualized survival and incidence curves for alternative management options improves satisfaction about cancer risk management decisions among women with BRCA1/2 mutations without increasing anxiety or changing management decisions. The benefit of the DSS is greatest among women with relatively low cancer-related anxiety at baseline. PMID- 16361632 TI - Role of imaging in pretreatment evaluation of early invasive cervical cancer: results of the intergroup study American College of Radiology Imaging Network 6651-Gynecologic Oncology Group 183. AB - PURPOSE: To compare magnetic resonance imaging (MRI) and computed tomography (CT) with each other and to International Federation of Gynecology and Obstetrics (FIGO) clinical staging in the pretreatment evaluation of early invasive cervical cancer, using surgicopathologic findings as the reference standard. PATIENTS AND METHODS: This prospective multicenter clinical study was conducted by the American College of Radiology Imaging Network and the Gynecologic Oncology Group from March 2000 to November 2002; 25 United States health centers enrolled 208 consecutive patients with biopsy-confirmed cervical cancer of FIGO stage > or = IB who were scheduled for surgery based on clinical assessment. Patients underwent FIGO clinical staging, helical CT, and MRI. Surgicopathologic findings constituted the reference standard for statistical analysis. RESULTS: Complete data were available for 172 patients; surgicopathologic findings were consistent with FIGO stages IA to IIA in 76% and stage > or = IIB in 21%. For the detection of advanced stage (> or = IIB), sensitivity was poor for FIGO clinical staging (29%), CT (42%), and MRI (53%); specificity was 99% for FIGO clinical staging, 82% for CT, and 74% for MRI; and negative predictive value was 84% for FIGO clinical staging, 84% for CT, and 85% for MRI. MRI (area under the receiver operating characteristic curve [AUC], 0.88) was significantly better than CT (AUC, 0.73) for detecting cervical tumors (P = .014). For 85% of patients, FIGO clinical staging forms were submitted after MRI and/or CT was performed. CONCLUSION: CT and MRI performed similarly; both had lower staging accuracy than in prior single-institution studies. Accuracy of FIGO clinical staging was higher than previously reported. The temporal data suggest that FIGO clinical staging was influenced by CT and MRI findings. PMID- 16361633 TI - Risk of epithelial ovarian cancer recurrence in patients with rising serum CA-125 levels within the normal range. AB - PURPOSE: To evaluate the risk of epithelial ovarian cancer (EOC) recurrence in patients with rising serum cancer antigen 125 (CA-125) levels that remain below the upper limit of normal (< 35 U/mL). PATIENTS AND METHODS: All patients treated for EOC between September 1997 and March 2003 were identified and screened retrospectively for the following: (1) elevated serum CA-125 at time of diagnosis, (2) complete clinical and radiographic response (CR) to initial treatment with normalization of serum CA-125, (3) at least three serial serum CA 125 determinations that remained within the normal range, and (4) clinical and/or radiographic determination of disease status at the time of last follow-up or recurrence. For statistical analyses, univariate regression models were used to compare absolute and relative changes in CA-125 levels among patients with recurrent disease and those without EOC recurrence. RESULTS: A total of 39 patients satisfied study inclusion criteria; 22 patients manifested EOC recurrence at a median interval from complete response of 11 months. The median follow-up time from complete response to last contact was 32 months for the 17 patients in the no recurrence group. A relative increase in CA-125 of 100% (odds ratio [OR] = 23.7; 95% CI, 2.9 to 192.5; P = .003) was significantly predictive of recurrence. From baseline CA-125 nadir levels, an absolute increase in CA-125 of 5 U/mL (OR = 8.4; 95% CI, 2.2 to 32.6; P = .002) and 10 U/mL (OR = 71.2; 95% CI, 4.8 to > 999.9; P = .002) were also significantly associated with the likelihood of concurrent disease recurrence. CONCLUSION: Among patients with EOC in complete clinical remission, a progressive low-level increase in serum CA-125 levels is strongly predictive of disease recurrence. PMID- 16361634 TI - Women with synchronous primary cancers of the endometrium and ovary: do they have Lynch syndrome? AB - PURPOSE: Lynch syndrome (hereditary nonpolyposis colorectal cancer; HNPCC) is an autosomal-dominant cancer predisposition syndrome that increases risk for multiple cancers, including colon, endometrial, and ovarian cancer. Revised Bethesda Criteria recommend that patients with two HNPCC-associated cancers undergo molecular evaluation to determine whether they have a mismatch repair (MMR) defect associated with HNPCC. The purpose of our study was to determine the likelihood of MMR defects (MSH2, MSH6, MLH1) in women with synchronous endometrial and ovarian cancer. PATIENTS AND METHODS: Between 1989 and 2004, 102 women with synchronous endometrial and ovarian cancers were identified; 59 patients had tumor blocks available for analysis. Patients were divided into risk groups based on family history: high (met Amsterdam criteria), medium (personal history or first-degree relative with an HNPCC-associated cancer), and low (all others). Protein expression for MSH2, MSH6, and MLH1 was evaluated by immunohistochemistry. Microsatellite instability and MLH1 promoter methylation analyses were performed on a subset of cases. RESULTS: Median age was 50 years. Two patients met Amsterdam criteria for HNPCC. Five additional patients, all medium-risk, had molecular findings consistent with a germline mutation of either MSH2 or MLH1. None of the low-risk patients had molecular results consistent with a germline mutation. CONCLUSION: Overall, 7% of women in our cohort met either clinical or molecular criteria for Lynch syndrome. All of these women had a prior history or a first-degree relative with an HNPCC-associated cancer. Limiting genetic evaluation to women with synchronous endometrial and ovarian cancer who have a family history suggestive of HNPCC may appropriately identify women with Lynch syndrome. PMID- 16361635 TI - Predictive utility of circulating methylated DNA in serum of melanoma patients receiving biochemotherapy. AB - PURPOSE: Currently, no validated blood-based assays accurately predict treatment response or outcome in melanoma patients. We hypothesized that methylation of tumor-related genes detected in serum DNA could predict disease outcome and therapeutic response in patients receiving concurrent biochemotherapy (BC) for metastatic melanoma. PATIENTS AND METHODS: American Joint Committee on Cancer stage IV melanoma patients (N = 50) had blood drawn before administration of BC. Patients (n = 47) were classified as BC responders or nonresponders. Responders (n = 23) demonstrated a complete or partial response following BC; nonresponders (n = 24) demonstrated progressive disease. Hypermethylation of Ras association domain family 1 (RASSF1A), retinoic acid receptor-beta2 (RAR-beta2), and O6 methylguanine DNA methyltransferase (MGMT) genes were assessed by methylation specific polymerase chain reaction. RESULTS: Circulating methylated RASSF1A was significantly less frequent for responders (three of 23 patients; 13%) than nonresponders (10 of 24 patients; 42%; P = .028). Patients with RASSF1A, RAR beta2, or at least one serum methylated gene had significantly worse overall survival than patients with no methylated genes (log-rank, P = .013, .021, and .01, respectively). Methylated RASSF1A was the only factor that significantly correlated with overall survival and BC response (risk ratio, 2.38; 95% CI, 1.16 to 4.86; P = .018; odds ratio = 0.21; 95% CI, 0.05 to 0.90; P = .036). CONCLUSION: Detection of circulating methylated DNA in serum can predict response to BC and disease outcome. PMID- 16361636 TI - Phase II study of imatinib mesylate plus hydroxyurea in adults with recurrent glioblastoma multiforme. AB - PURPOSE: We performed a phase II study to evaluate the combination of imatinib mesylate, an adenosine triphosphate mimetic, tyrosine kinase inhibitor, plus hydroxyurea, a ribonucleotide reductase inhibitor, in patients with recurrent glioblastoma multiforme (GBM). PATIENTS AND METHODS: Patients with GBM at any recurrence received imatinib mesylate plus hydroxyurea (500 mg twice a day) orally on a continuous, daily schedule. The imatinib mesylate dose was 500 mg twice a day for patients on enzyme-inducing antiepileptic drugs (EIAEDs) and 400 mg once a day for those not on EIAEDs. Assessments were performed every 28 days. The primary end point was 6-month progression-free survival (PFS). RESULTS: Thirty-three patients enrolled with progressive disease after prior radiotherapy and at least temozolomide-based chemotherapy. With a median follow-up of 58 weeks, 27% of patients were progression-free at 6 months, and the median PFS was 14.4 weeks. Three patients (9%) achieved radiographic response, and 14 (42%) achieved stable disease. Cox regression analysis identified concurrent EIAED use and no more than one prior progression as independent positive prognostic factors of PFS. The most common toxicities included grade 3 neutropenia (16%), thrombocytopenia (6%), and edema (6%). There were no grade 4 or 5 events. Concurrent EIAED use lowered imatinib mesylate exposure. Imatinib mesylate clearance was decreased at day 28 compared with day 1 in all patients, suggesting an effect of hydroxyurea. CONCLUSION: Imatinib mesylate plus hydroxyurea is well tolerated and associated with durable antitumor activity in some patients with recurrent GBM. PMID- 16361637 TI - MYC amplification and polysomy 8 in chondrosarcoma: array comparative genomic hybridization, fluorescent in situ hybridization, and association with outcome. AB - PURPOSE: To identify recurrent regions of genomic gain or loss in chondrosarcoma in a clinically relevant and statistically valid fashion. MATERIALS AND METHODS: Array comparative genomic hybridization (CGH) results of 15 frozen tumor samples of high-grade chondrosarcoma for chromosome 8 are presented. A separate subset of 116 cartilaginous tumors with outcome data was used for validation. RESULTS: Array CGH identified gain at 8q24.12-q24.13, the region of the MYC (c-Myc) oncogene, as a frequent change in high-grade chondrosarcoma. In the validation arm of 116 cartilaginous tumors, MYC was frequently amplified in G2 (15%), G3 (20%), and dedifferentiated (21%) chondrosarcomas. No amplification was identified in samples of enchondroma and grade 1 chondrosarcoma. In samples without MYC amplification, polysomy 8 was a frequent finding in grade 1 (18%), grade 2 (31%), grade 3 (80%), and dedifferentiated (29%) chondrosarcomas, but was not found in any samples of enchondroma. MYC protein expression was identified in all samples with amplification, but was also frequent in the remaining samples without amplification or polysomy 8. Kaplan-Meier survival curves for overall survival showed a statistically significant difference for patients with MYC amplification or polysomy 8 (P = .034). Univariate analysis involving Cox proportional hazards models showed that grade (P = .003), polysomy 8 (P = .045), and MYC amplification (P = .053) correlated with shorter overall survival. By multivariate analysis, grade of chondrosarcoma (P = .026) was the only factor to reach statistical significance. CONCLUSION: MYC amplification and polysomy 8 can be used as markers of prognostic importance in chondrosarcoma. Molecular targeting of MYC expression may have therapeutic potential in the future for subsets of chondrosarcoma. PMID- 16361638 TI - Efficacy and safety analysis of epoetin alfa in patients with small-cell lung cancer: a randomized, double-blind, placebo-controlled trial. AB - PURPOSE: This randomized, double-blind, placebo-controlled trial (N93-004) evaluated the effects of epoetin alfa on tumor response to chemotherapy and survival in patients with small-cell lung cancer (SCLC). PATIENTS AND METHODS: Adult patients with hemoglobin < or = 14.5 g/dL starting chemotherapy received epoetin alfa 150 U/kg or placebo subcutaneously 3 times weekly until 3 weeks after completion of chemotherapy. Survival was assessed for 3 years. The primary end point was the proportion of patients with complete or partial response after three chemotherapy cycles. RESULTS: The trial was terminated prematurely after 224 of a projected 400 patients were accrued. Baseline characteristics were similar between groups. Epoetin alfa and placebo patients (n = 109 and n = 115, respectively) had mean baseline hemoglobin of 12.8 g/dL and 13.0 g/dL, respectively. Overall tumor response was similar between the epoetin alfa and placebo groups after three chemotherapy cycles (72% and 67%, respectively; 95% CI of difference, -6% to 18%) and after completion of chemotherapy (60% and 56%, respectively; 95% CI of difference, -9% to 17%). Epoetin alfa and placebo groups had similar median overall survival (10.5 and 10.4 months, respectively) and overall mortality (91.7% and 87.8%, respectively; hazard ratio, 1.172; 95% CI, 0.887 to 1.549; P = .264). Hemoglobin was maintained in the prechemotherapy range in epoetin alfa patients, but decreased substantially in placebo patients. Fewer epoetin alfa patients than placebo patients required transfusion. CONCLUSION: These results suggest that in newly diagnosed patients with SCLC epoetin alfa does not affect tumor response to chemotherapy or survival. However, the early trial closure makes these conclusions preliminary. PMID- 16361639 TI - Targeted induction of apoptosis in cancer management: the emerging role of tumor necrosis factor-related apoptosis-inducing ligand receptor activating agents. AB - Targeted induction of programmed cell death or apoptosis via the extrinsic apoptotic pathway represents an unexploited therapeutic strategy to destroy cancer cells. The activation of cell surface receptors by the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) results in direct stimulation of apoptotic signaling pathways (extrinsic stimulation). Molecules that directly activate these receptors, such as agonistic monoclonal antibodies to the TRAIL receptors and recombinant TRAIL, are being developed as monotherapies and as part of combination therapies with existing chemotherapeutic drugs and other therapeutic modalities. This article examines the TRAIL receptors as potential targets for activating the TRAIL-mediated apoptosis pathway and presents the current status of novel therapeutics that exploit this pathway, particularly focusing on agonistic monoclonal antibodies to the TRAIL receptors. The preclinical activity, the status of ongoing evaluations, and the potential clinical impact of these novel agents are reviewed. PMID- 16361640 TI - Targeting the cell cycle: a new approach to cancer therapy. AB - The cell cycle represents a series of tightly integrated events that allow the cell to grow and proliferate. Critical parts of the cell cycle machinery are the cyclin-dependent kinases (CDKs), which, when activated, provide a means for the cell to move from one phase of the cell cycle to the next. The CDKs are regulated positively by cyclins and regulated negatively by naturally occurring CDK inhibitors (CDKIs). Cancer represents a dysregulation of the cell cycle such that cells that overexpress cyclins or do not express the CDKIs continue to undergo unregulated cell growth. The cell cycle also serves to protect the cell from DNA damage. Thus, cell cycle arrest, in fact, represents a survival mechanism that provides the tumor cell the opportunity to repair its own damaged DNA. Thus, abrogation of cell cycle checkpoints, before DNA repair is complete, can activate the apoptotic cascade, leading to cell death. Now in clinical trials are a series of targeted agents that directly inhibit the CDKs, inhibit unrestricted cell growth, and induce growth arrest. Recent attention has also focused on these drugs as inhibitors of transcription. In addition, there are now agents that abrogate the cell cycle checkpoints at critical time points that make the tumor cell susceptible to apoptosis. An understanding of the cell cycle is critical to understanding how best to clinically develop these agents, both as single agents and in combination with chemotherapy. PMID- 16361641 TI - Symptoms in cancer patients and an unusual tumor: Case 1. Regression of hypertrophic pulmonary osteoarthropathy following chemotherapy for lung metastases of a nasopharyngeal carcinoma. PMID- 16361642 TI - Symptoms in cancer patients and an unusual tumor: Case 2. Docetaxel-related ischemic colitis. PMID- 16361643 TI - Symptoms in cancer patients and an unusual tumor: Case 3. Follicular dendritic cell sarcoma. PMID- 16361644 TI - Calculation of survival of patients with stage III melanoma. PMID- 16361645 TI - Misleading statements in industry-sponsored meta-analysis of itraconazole. PMID- 16361646 TI - Bone loss with exemestane: Is the jury still out? PMID- 16361647 TI - Treatment guidelines for adjuvant breast cancer are moving toward double standards: one for the rich and one for the poor. PMID- 16361648 TI - Validation of microarray data by quantitative reverse-transcriptase polymerase chain reaction. PMID- 16361649 TI - Overall survival of patients with advanced colorectal cancer correlates with availability of fluorouracil, irinotecan, and oxaliplatin regardless of whether doublet or single-agent therapy is used first line. PMID- 16361650 TI - Time to re-evaluate sentinel node biopsy in melanoma post-multicenter selective lymphadenectomy trial. PMID- 16361651 TI - Randomized phase II studies and their ethics. PMID- 16361652 TI - Lens extraction in primary angle closure. PMID- 16361653 TI - Prevention of diabetic blindness. PMID- 16361654 TI - Building a safer trabeculectomy. PMID- 16361655 TI - Thrombospondin in the eye. PMID- 16361656 TI - Cataract care is mobile. PMID- 16361657 TI - Willingness to pay for cataract surgery in two regions of Tanzania. AB - BACKGROUND: Knowing what rural populations are willing to pay for cataract surgery is essential if improvements in cost recovery in eye care service provision programmes are to take place. The authors sought to learn about willingness to pay for cataract surgery in two separate regions of Tanzania. METHODS: Patients desiring cataract surgery were interviewed in Kilimanjaro Region and Iringa Region of Tanzania to learn how much they and their families were willing to pay for surgery and how "wealthy" (using ownership of several household objects as a proxy for wealth) the household was. RESULTS: 60 cataract patients in Kilimanjaro and 49 in Iringa were interviewed. "Wealth" was significantly associated with willingness to pay in each region. The average expressed willingness to pay was 2457Tsh (SD 4534) or approximately $US2.30, which is far below the actual cost of providing the service. CONCLUSION: There were significant differences in the expressed willingness to pay between Iringa and Kilimanjaro patients, which may reflect differences in the services provided in the regions. Willingness to pay may increase as the population gains familiarity and trust in the service. It may also be increased by ensuring that pricing is uniform and clearly advertised throughout regions and by educating health workers and counselling patients about the real costs of providing high quality surgery. Offering "free" services to all may result in lower expressed willingness to pay. PMID- 16361658 TI - Visual acuity after acute primary angle closure and considerations for primary lens extraction. AB - BACKGROUND/AIM: Primary lens extraction has been advocated for acute primary angle closure (APAC), but it is not known if this is warranted in all cases. The aim of this study was to investigate the visual acuity (VA) of APAC eyes shortly after resolution of the acute episode in order to assess the appropriateness of performing such surgery in this condition. METHODS: This was a prospective observational case series. As part of a randomised controlled trial comparing phacoemulsification and laser iridotomy, 135 consecutive APAC subjects over a 2 year period underwent subjective refraction and measurement of Snellen VA once the acute episode had resolved with reduction of intraocular pressure (IOP) and improved corneal clarity. RESULTS: Subjects were predominantly Chinese (95.6%) and female (79.3%), with a mean age of 63.6 (SD 9.6) years. When assessed 1.7 (2.7) days after presentation, the majority of APAC cases (50.4%) had good VA (6/12 or better), with more than a quarter of cases having VA of 6/7.5 or better. Poor VA was associated with duration of symptoms (p = 0.04, OR = 4.1, 95% CI 1.1 to 15.7) and time taken to resolution of APAC (p = 0.04, OR = 2.2, 95% CI 1.02 to 4.6), but not with sex (p = 0.31), age (p = 0.26), duration from presentation to measurement of visual acuity (p = 0.53), or presenting IOP (p = 0.73). CONCLUSION: Within days after APAC, more than half of APAC affected eyes had good VA (6/12 or better). The role of lens extraction in the management of APAC warrants further debate, especially for eyes with good VA. PMID- 16361659 TI - Scanning laser entoptic perimetry for the detection of visual defects associated with diabetic retinopathy. AB - AIM: To determine the sensitivity and specificity of entoptic perimetry for diagnosing diabetic retinopathy at all levels of severity. METHODS: A prospective clinical study at the Shiley Eye Center, University of California, and San Diego. 30 patients with photographically documented diabetic retinopathy and 24 controls with a similar age distribution. Sensitivity and specificity of entoptic perimetry were computed for detecting clinically significant macular oedema within the central 120 degree radius of the fovea compared to fundus photographs. RESULTS: Entoptic perimetry can detect clinically significant diabetic retinopathy with a sensitivity of 0.88 and specificity of 1.00. Entoptic perimetry can detect the earliest stages of diabetic retinopathy with a sensitivity of 0.86. CONCLUSION: Scanning laser entoptic perimetry is an effective tool for detecting visual function loss caused by diabetic retinopathy. PMID- 16361660 TI - Five year follow up of laser in situ keratomileusis for all levels of myopia. AB - AIMS: To assess the long term refractive and visual outcome of patients who have laser in situ keratomileusis (LASIK) surgery. METHODS: This was a retrospective study of visual and refractive outcome of patients who had LASIK surgery performed in 1998 and 1999. All levels of myopia were included in the study. 49 patients attended for follow up. The main outcome measures were safety, predictability, efficacy, and stability. Postoperative complications and aberrations were also recorded. The mean preoperative spherical equivalent was 4.85. RESULTS: At 2 months postoperatively 67% of eyes were within plus or minus 0.5D of attempted correction with 81% within plus or minus 1.0D. At 5 years postoperatively 60% of eyes were within plus or minus 0.5D of attempted correction with 83% within plus or minus 1.0D. 88% of eyes had a vision of 6/12 or better at 2 months compared to 89% of eyes at 5 years. Best spectacle corrected visual acuity (BSCVA) was unchanged or improved in 51%. No eye lost more than one line of BSCVA. Overall, there was regression towards myopia with a mean change in refraction of -0.5D over the 5 years. As expected, severely myopic patients regressed more with a mean change of -1.06D. However, there was a high level of patient satisfaction with the surgery. CONCLUSION: LASIK surgery offers predictable results in terms of refractive and visual outcome with mild regression in refraction over time. PMID- 16361661 TI - Circadian variations in central corneal thickness and intraocular pressure in patients with glaucoma. AB - AIM: To analyse 24 hour variations in intraocular pressure (IOP) and central corneal thickness (CCT) in a group of glaucomatous patients. METHODS: 30 patients with primary open angle glaucoma were hospitalised and underwent circadian evaluations (at 8 pm, midnight, 4 am, 8 am, noon, and 4 pm) of supine and sitting IOP, respectively, measured using a Perkins and a Goldmann tonometer, and CCT measured using an ultrasonic pachymeter (the mean value of three measurements within 5 mum). All patients were treated with timolol 0.5% twice daily and latanoprost 0.005% once daily. RESULTS: Mean supine IOP was 15.3 (SD 3.7) mm Hg (range 10-25), with circadian fluctuations of 7.3 (3.3) mm Hg. Mean sitting IOP was 15.1 (3.9) mm Hg (range 8-26), with circadian fluctuations of 5.4 (3.1) mm Hg. Mean CCT was 534 (39) microm (range 443-637 microm) with circadian fluctuations of 16.5 (6.2) microm (range 6-31 microm). Both the within patient and within time point fluctuations in CCT were statistically significant (p<0.0001, ANOVA). CONCLUSIONS: The authors found considerable fluctuations in 24 hour IOP. The circadian fluctuations in CCT were small and, although statistically significant, did not seem to interfere with the circadian IOP assessment. PMID- 16361662 TI - Age related macular degeneration and sun exposure, iris colour, and skin sensitivity to sunlight. AB - BACKGROUND/AIM: It has been suggested that sun exposure may be a risk factor for age related macular degeneration (AMD) and that skin sensitivity to sunlight and iris colour could be confounding factors. The aim was to investigate this further in the white population. METHODS: 446 cases with end stage AMD were compared with 283 spouse controls. Data on sun exposure, places of residence, iris colour, subjective assessment of change in iris colour, hair colour at age 20, and skin sensitivity were obtained using a questionnaire. Iris colour was graded clinically by comparison with standard photographs. AMD was graded using stereoscopic colour fundus photographs as well as clinical examination and was defined as the presence of geographic atrophy or choroidal neovascularisation. All variables were included in a multiple logistic regression model including age, sex, and smoking. RESULTS: There was no association between AMD and sun exposure or related factors except for the suggestion of an association between sunburn prone skin type and geographic atrophy which reached borderline significance. CONCLUSIONS: No significant association between AMD and sun exposure, iris colour, change in iris colour, or hair colour was demonstrated. PMID- 16361663 TI - Factors predicting outcome of vitrectomy for diabetic macular oedema: results of a prospective study. AB - AIM: To determine preoperative demographic, clinical, and optical coherence tomography (OCT) factors which might predict the visual and anatomical outcome at 1 year in patients undergoing vitrectomy and inner limiting membrane peel for diabetic macular oedema (DMO). METHODS: A prospective, interventional case series of 33 patients who completed 1 year follow up. Measurements were taken preoperatively and at 1 year. Outcome measures were logMAR visual acuity (VA) and OCT macular thickness. A priori explanatory variables included baseline presence of clinical and/or OCT signs suggesting macular traction, grade of diabetic maculopathy, posterior vitreous detachment, fluorescein leakage and ischaemia on angiography, presence of subretinal fluid, and peroperative indocyanine green (ICG) use. RESULTS: 33 patients completed 1 year follow up. On average VA deteriorated by 0.035 logMAR (p = 0.40). Macular thickness significantly improved by a mean of 139 microm (95% CI; 211 to 67, p<0.001). Patients with evidence of clinical and/or OCT macular traction significantly improved logMAR acuity (logMAR improvement = 0.08) compared with patients without traction (logMAR deterioration 0.11, p = 0.01). Presence of subretinal fluid significantly predicted worse postoperative result (p = 0.01) CONCLUSION: On average, patients showed a statistically significant improvement in central macular thickness following treatment but a marginal acuity worsening. Presence of subretinal fluid on OCT is hypothesised to be exudative rather than tractional in nature. The visual benefit of vitrectomy for DMO in this study was limited to patients who exhibit signs of macular traction either clinically and/or on OCT. PMID- 16361664 TI - Cystoid macular oedema in paediatric aphakia and pseudophakia. AB - AIM: To assess the incidence of cystoid macular oedema (CMO), following lensectomy, anterior vitrectomy (limbal and pars plicata), and primary posterior capsulorhexis following surgery for congenital and developmental cataract. METHODS: A prospective study was carried out involving 30 eyes (20 children) with cataract. Lensectomy, primary posterior capsulorhexis, and anterior vitrectomy were performed on all eyes. Vitrectomy was performed using either a limbal or pars plicata approach. Fluorescein angiography was carried out 4-7 weeks after surgery. Fluorescein was administered intravenously and visualised using the Retcam. RESULTS: Anterior vitrectomy was carried out via the limbal approach in 12 eyes and, of these, six were implanted with an intraocular lens. The pars plicata approach was carried out in 18 eyes and 10 received an implant. CMO was not detected in any eye. CONCLUSION: CMO was not detected in the early postoperative period irrespective of approach to anterior vitrectomy or presence of an intraocular lens. Intravenous fluorescein angiography was performed without complication. PMID- 16361665 TI - In the absence of strabismus what constitutes a visual deficit in children? AB - AIM: To examine the range of monocular crowded logMAR acuity and cycloplegic refraction of children aged 3 years and 4 years without strabismus. METHODS: Data were collected prospectively. All children had full orthoptic and ophthalmic examination. Inclusion principally required the absence of strabismus, symptoms, and ocular pathology. Criteria were used to categorise the refraction findings as normal, borderline, or abnormal. RESULTS: 118 children aged 3 years (mean 3 years 6 months (SD 2 months) and 67 children aged 4 years (mean 4 years 4 months (SD 3 months) fulfilled the inclusion criteria. For the children with normal refraction, the mean monocular logMAR acuity was 0.200 (SD 0.09) for 3 year olds (n = 84) and 0.140 (SD 0.08) for 4 year olds (n = 45); the mean interocular acuity difference (IAD) was 0.03 (SD 0.06) for 3 year olds and 0.03 (SD 0.04) for 4 year olds. The 95% confidence limits were calculated for monocular acuity and IAD and compared to the acuity scores of the children with borderline (n = 17) and abnormal refractive error (n = 38). 15 borderline and 16 abnormal cases had a visual acuity within these limits. CONCLUSION: A wide range of refraction may be associated with a good level of visual acuity in children aged 3 years and 4 years. PMID- 16361666 TI - Safe trabeculectomy technique: long term outcome. AB - AIM: To assess the long term outcome of a new trabeculectomy technique. METHODS: Trabeculectomy was performed using a fornix based conjunctival flap, an anterior chamber maintainer, a standardised punch technique, and a combination of adjustable and releasable sutures in 56 eyes of 53 patients. The main outcome measures were the postoperative intraocular pressure (IOP) and the frequency of early postoperative complications. The mean follow up time was 15.7 (range 12-21) months. RESULTS: The mean preoperative and postoperative IOP at 12 months were 21.2 (SD 6) and 12.8 (3.0) mm Hg, respectively. All patients had an IOP of <21 mm Hg, 90.9% had an IOP <18 mm Hg, and 61.4% had an IOP <14 mm Hg. Postoperative complications were infrequent: flat anterior chamber (1.8%), bleb leakage (0%), or hypotony (1.5%) beyond 3 weeks, or choroidal detachment at any time point (8.9%). CONCLUSIONS: This novel trabeculectomy method offers the possibility to tailor the IOP postoperatively with a minimum of postoperative complications and excellent IOP control at the long term follow up. PMID- 16361667 TI - Impaired expression of thrombospondin-1 in eyes with age related macular degeneration. AB - AIMS: This study investigated the expression and localisation of thrombospondin-1 (TSP-1), a known anti-angiogenic extracellular matrix protein, in normal aged control human eyes and eyes with age related macular degeneration (AMD). METHODS: Immunohistochemical analysis with mouse anti-human TSP-1 antibody and mouse anti human CD 34 antibody, as a blood vessel marker, was performed on frozen sections from macular and peripheral blocks of aged control donor eyes (n = 12; mean age 78.8 years), and eyes with AMD (n = 12; mean age 83.9 years). Pigment in retinal pigment epithelium (RPE) and choroidal melanocytes was bleached. Three independent observers scored the immunohistochemical reaction product. RESULTS: In the macular region, TSP-1 expression was observed intensely in Bruch's membrane and weakly in RPE basement membrane, choriocapillaris, and the wall of large choroidal blood vessels in the aged control eyes. In eyes with AMD, TSP-1 immunoreactivity was significantly lower in all structures except RPE basement membrane (p<0.01). There was significantly lower TSP-1 in the far periphery than the equator and submacular regions in all eyes. TSP-1 immunoreactivity was low in choroidal neovascularisation (CNV), but it was high and diffuse in adjacent scar tissue. CONCLUSION: These findings suggest that decreased TSP-1 in Bruch's membrane and choroidal vessels during AMD may permit the formation of CNV. PMID- 16361668 TI - Epithelial proliferative potential of organ cultured corneoscleral rims; implications for allo-limbal transplantation and eye banking. AB - AIMS: To determine the epithelial proliferative capacity of organ cultured limbal tissue and correlate this with various donor and eye banking factors. METHODS: 24 corneoscleral limbal (CSL) rims left over from penetrating keratoplasty were split in half and set up as in vitro explant cultures. Corneal epithelial proliferative potential (CEPP) was assessed by the number of "cycles" of growth achieved before explants underwent exhaustion and failure to generate an epithelium to subconfluence. The dependence of CEPP on the age of the donor, time of death to enucleation, time of enucleation to organ culture, and time in organ culture in the eye bank was determined. RESULTS: CSL rims were capable of up to four cycles of culture with a wide variation between tissue samples. Of the various factors examined, death to enucleation time was the only statistically significant factor affecting the CEPP (regression coefficient: -0.062 (cycles/hour), CI -0.119 to -0.004, p = 0.037). Time in organ culture had little effect on CEPP. CONCLUSIONS: Preselected organ cultured CSL rims from eye banks may offer a viable alternative tissue source for use in allo-limbal transplantation. PMID- 16361669 TI - Longitudinal glaucoma screening for siblings of patients with primary open angle glaucoma: the Nottingham Family Glaucoma Screening Study. AB - AIMS: To assess the prevalence and cumulative incidence of open angle glaucoma (OAG) in a cohort group of siblings of OAG probands. METHODS: Between 1994 and 2003, a group of siblings of OAG probands underwent both initial and follow up standardised ophthalmic examinations. Siblings were classified as "definite glaucoma" (primary OAG (POAG) and normal tension glaucoma (NTG)), "glaucoma suspects" (NTG suspects or ocular hypertension (OHT)), and normal. The prevalence and cumulative incidence of OAG over the follow up interval were calculated. RESULTS: At the initial study, 271 siblings (mean age 63.6 years; female to male ratio 1.2) from 156 probands were examined. 32 (11.8%) were classified as definite glaucoma and 15 (5.5%) as suspects. In the follow up study, 157 of the 224 "normal" siblings from the initial study were examined (mean interval from initial study 7.0 (SD 1.0) years). 11 (7%) were classified as definite glaucoma and 30 (19.1%) as suspects. There were significant trends of increasing prevalence and incidence of OAG with age and a lifetime risk estimated at approximately 20% by age 70. CONCLUSION: Siblings of glaucoma patients have an increased risk of developing glaucoma and the risk increases with age. An effective and repeated screening programme should be considered for this high risk group. PMID- 16361670 TI - Ultrasound biomicroscopy of Chinese eyes with iridocorneal endothelial syndrome. AB - AIM: To document the ultrasound biomicroscopic (UBM) findings in Chinese patients with iridocorneal endothelial (ICE) syndrome. METHODS: 21 patients with ICE syndrome and 15 normal subjects underwent UBM. UBM findings of anterior segment were compared between normal subjects and three clinical types of ICE syndrome: progressive iris atrophy (PIA), Chandler's syndrome (CS), and Cogan-Reese syndrome (CRS). RESULTS: Central anterior chamber depth was significantly less in patients with ICE syndrome (2.25 (SD 0.32) mm) than in normal subjects (2.76 (0.32) mm). Peripheral anterior synechiae were observed in all the ICE patients by UBM. Three out of four CRS subjects showed an "arborised" shape of iridocorneal angle. Two eyes out of 10 with CS presented bridge-shaped synechiae. A membrane-like mound was observed in iridocorneal angle in two patients: one with CRS and one with CS. UBM was found to be more effective in detecting peripheral anterior synechiae (PAS) and iris atrophy than slit lamp microscopy and gonioscopy, mainly because of corneal oedema in patients with CS. Four out of 11 patients with unilateral ICE syndrome had shallow or closed anterior chamber angles in their fellow eyes. Two of them successfully responded to laser peripheral iridotomy. CONCLUSIONS: UBM is an effective method to reveal the anterior segment features and provides a useful tool in the diagnosis of ICE syndrome. Different subtypes of ICE syndrome may have different UBM manifestations. UBM can help to identify angle closure in the fellow eye of unilateral ICE syndromes. PMID- 16361671 TI - Topography of the frequency doubling perimetry visual field compared with that of short wavelength and achromatic automated perimetry visual fields. AB - BACKGROUND: Traquair described the topography of visual field sensitivity as a "hill" or "island" of vision. Achromatic automated perimetry (AAP) demonstrates this shape of the visual field in photopic conditions. Techniques claimed to target the magnocellular pathway (frequency doubling perimetry, FDP) and those using a stimulus targeting the koniocellular pathway (short wavelength (or blue on yellow) automated perimetry, SWAP), might produce one that is different. The authors compared the visual field topography from FDP with those of SWAP and AAP, to investigate whether there were significant differences in their shape. METHOD: A sample of 51 patients with previously confirmed normal perimetry were recruited; either low risk glaucoma suspects or normal controls. AAP, SWAP, and FDP perimetry was performed in random order on the same day. The topography of each field was analysed to determine its average shape and to compare results in the same individuals. RESULTS: The topography of the visual field produced by each perimeter differed significantly. While all three had maximal sensitivity centrally, over the 24 degrees from the centre to the periphery, mean sensitivities decreased by 4.9 decibels (dB) for AAP and 7.3 dB for SWAP, while FDP sensitivities by just 1.8 dB over 20 degrees (the extent of the FDP field). FDP mean sensitivities decreased by approximately 0.3 dB with every 10 year increase in age, compared with 1 dB for AAP and 2 dB for SWAP. CONCLUSION: While the topography of the SWAP (koniocellular) field is steeper than corresponding AAP fields, that of the FDP (magnocellular) visual field was considerably flatter. The difference in this shape may reflect retinotopic or cortical mechanisms, which are specific to the magnocellular pathways. PMID- 16361672 TI - Smoking and age related macular degeneration: the number of pack years of cigarette smoking is a major determinant of risk for both geographic atrophy and choroidal neovascularisation. AB - BACKGROUND/AIMS: There is evidence that smoking is a risk factor for age related macular degeneration (AMD). However, not all studies have demonstrated this association and several key questions about the role of smoking in AMD have still to be determined. The aim of this study was to further investigate this relation for both choroidal neovascularisation (CNV) and geographic atrophy (GA). METHODS: To investigate the relation between smoking and the risk of developing age related macular degeneration (AMD) in white people, 435 cases with end stage AMD were compared with 280 controls. All subjects had graded stereoscopic colour fundus photography and AMD was defined as the presence of GA or CNV. Smoking history was assessed using multiple parameters in a detailed questionnaire. RESULTS: Comparison of current and former smokers with non-smokers was consistent with smoking being a risk factor for AMD but did not reach statistical significance. There was a strong association between AMD and pack years of cigarette smoking (p = 0.002), the odds ratio increasing with the amount smoked; for subjects with more than 40 pack years of smoking the odds ratio was 2.75 (95% CI 1.22 to 6.20) compared with non-smokers. Both types of AMD showed a similar relation; smoking more than 40 pack years of cigarettes was associated with an odds ratio of 3.43 (95% CI 1.28 to 9.20) for GA and 2.49 (95% CI 1.06 to 5.82) for CNV. Stopping smoking was associated with reduced odds of AMD and the risk in those who had not smoked for over 20 years was comparable to non-smokers. The risk profile was similar for males and females. Passive smoking exposure was associated with an increased risk of AMD (OR 1.87; 95% CI 1.03 to 3.40) in non smokers. CONCLUSIONS: The authors have demonstrated a strong association between the risk of both GA and CNV and pack years of cigarette smoking. This provides support for a causal relation between smoking and AMD. They also show an increased risk for AMD in non-smokers exposed to passive smoking. Stopping smoking appears to reduce the risk of developing AMD. PMID- 16361673 TI - Molecular pathology of X linked retinoschisis: mutations interfere with retinoschisin secretion and oligomerisation. AB - BACKGROUND/AIM: X linked retinoschisis (XLRS) is caused by mutations in RS1 which encodes the discoidin domain protein retinoschisin, secreted by photoreceptors and bipolar cells. Missense mutations occur throughout the gene and some of these are known to interfere with protein secretion. This study was designed to investigate the functional consequences of missense mutations at different locations in retinoschisin. METHODS AND RESULTS: The authors developed a structural model of the retinoschisin discoidin domain and used this to predict the effects of missense mutations. They expressed disease associated mutations and found that those affecting conserved residues prevented retinoschisin secretion. Most of the remaining mutations cluster within a series of loops on the surface of the beta barrel structure and do not interfere with secretion, suggesting this region may be a ligand binding site. They also demonstrated that wild type retinoschisin octamerises and associates with the cell surface. A subgroup of secreted mutations reduce oligomerisation (C59S, C219G, C223R). CONCLUSIONS: It is suggested that there are three different molecular mechanisms which lead to XLRS: mutations interfering with secretion, mutations interfering with oligomerisation, and mutations that allow secretion and oligomerisation but interfere with retinoschisin function. The authors conclude that binding of oligomerised retinoschisin at the cell surface is important in its presumed role in cell adhesion. PMID- 16361674 TI - Contrast sensitivity in 10 year old preterm and full term children: a population based study. AB - AIMS: To determine the contrast sensitivity (CS) in 10 year old prematurely born children, previously included in a population based study on the incidence of retinopathy of prematurity (ROP), and in full term controls. METHODS: This study included 205 prematurely born children and 215 children born at term, from the same geographical area and study period. CS was assessed monocularly with the Vistech 6500 test at five spatial frequencies (1.5-18 cycles/deg). RESULTS: Prematurely born children had statistically significant lower CS at all frequencies, compared to full term ones. The mean differences in logarithmic CS were 0.03 (1.5 cycles/deg), 0.09 (3 cycles/deg), 0.10 (6 cycles/deg), 0.12 (12 cycles/deg), and 0.19 (18 cycles/deg). Even when the children with ROP and neurological disorders were excluded there was a difference between the two groups. Children who had been treated with cryotherapy had the lowest CS. CONCLUSION: CS was lower in 10 year old prematurely born children than in full term ones of the same age. Whether this finding affects their visual function in daily life is uncertain. PMID- 16361675 TI - Effect of age on adult stereoacuity as measured by different types of stereotest. AB - AIM: To examine how stereoacuity changes with age as measured by a variety of stereotests. METHODS: Stereoacuity has been measured in 60 normal subjects aged 17-83 years by a single observer using TNO, Titmus, Frisby near, and Frisby-Davis distance stereotests. Motor fusion was measured at (1/3) metre and 6 metres. RESULTS: Overall stereoacuity measured by all tests showed a mild decline with age (p<0.001 for all tests; Spearman rank correlation). A marked reduction to screening or absent levels of stereoacuity was seen in five subjects aged over 55, but only with the TNO stereotest. All these subjects were able to achieve a stereoacuity of 200 seconds of arc or better with the Titmus test and 340 seconds of arc or better using the Frisby near stereotest. There was a small decline with age in the base out motor fusion range measured at 6 metres (p<0.05; Spearman rank correlation). No subject described difficulty in judging distances for everyday tasks. CONCLUSIONS: Although subjects showed some decline in stereoacuity with age by all tests, the large drop in stereoacuity seen in some older subjects using the TNO test was probably due to difficulty overcoming the dissociative effect of the test rather than a true reduction in cortical disparity detection. Results of random dot stereotests should be interpreted with caution in older patients, particularly with respect to their ability to perform everyday visual tasks. PMID- 16361676 TI - Expression of glutamine synthetase and cell proliferation in human idiopathic epiretinal membrane. AB - BACKGROUND/AIM: The mechanisms of the cellular origin and cell proliferation in the idiopathic epiretinal membrane (ERM) are unsolved. The aim of this study was to examine the expression of cell cycle related molecules and glutamine synthetase (GS), which is expressed in Muller cells and their processes, in ERM tissues. METHODS: The ERMs were surgically removed using pars plana vitrectomy. Formalin fixed, paraffin embedded ERM tissues were analysed by immunohistochemistry with anti-cyclin D1, p27 (KIP1), proliferating cell nuclear antigen (PCNA), and GS antibodies. RESULTS: The histopathological findings showed that all the ERMs consisted of oval or spindle mononuclear cells with thin collagen-like tissues. Immunoreactivity for GS was detected in collagen-like tissues of ERM, presenting a continuous, isodense pattern. GS immunopositive cells in all cases expressed PCNA in their nuclei. Nuclear immunoreactivity for cyclin D1 was noted in the ERM constituent cells, whereas p27 (KIP1) positive nuclei were not detected. CONCLUSION: Cyclin D1 and PCNA were expressed in the idiopathic ERM, which was mainly derived from Muller cells and extensions of their processes. PMID- 16361677 TI - Retinal ganglion cells toxicity caused by photosensitising effects of intravitreal indocyanine green with illumination in rat eyes. AB - AIM: To investigate the effects of indocyanine green (ICG) with or without illumination on rat retinal ganglion cells (RGC) and retinal morphology. METHODS: Intravitreal injections of 1.0 mg/ml ICG solution were performed in rat eyes with or without subsequent illumination for 5 minutes. Eyes in the control group had intravitreal injections of balanced salt solution with illumination. Retrograde labelling of RGC with 6% Fluoro-Gold was performed 1 month later and RGC densities were compared between the three groups. Light microscopy with measurements of outer nuclear layer (ONL) and inner nuclear layer (INL) thicknesses were also performed and compared. RESULTS: Eyes with ICG without illumination showed insignificant reduction in RGC density compared with the control group (p = 0.28), whereas a significant decrease in RGC density was found in eyes that had ICG injection with illumination (p = 0.036). A significant increase in ONL thickness was also observed in the ICG with illumination treated eyes compared with the ICG without illumination and the control groups (p<0.001). No significant difference in INL thickness was observed between the three groups. CONCLUSIONS: Intravitreal injection of 0.1 mg/ml ICG in rat eyes followed by illumination resulted in photosensitising toxicity to RGC. Lower ICG concentration or illumination level should be considered when performing ICG assisted macular surgery. PMID- 16361678 TI - Toll-like receptors in ocular immunity and the immunopathogenesis of inflammatory eye disease. AB - Microbial agents have an important role in the pathogenesis of various inflammatory eye diseases, such as uveitis and keratitis. Microbial infections of the eye such as microbial keratitis, ocular onchocerciasis, bacterial endophthalmitis, viral retinitis, and other infectious uveitis are unfortunately common. In addition, microbial agents have been implicated in the pathogenesis of "non-infectious" immune mediated diseases such as HLA-B27 associated acute anterior uveitis. Toll-like receptors (TLR) are a family of pattern recognition receptors that initiates rapid host innate immune response to microbial components known as pathogen associated molecular patterns, which are unique to a given class of microbes, such as lipopolysaccharide of Gram negative bacteria. Recent in vitro and in vivo studies have demonstrated the expression and function of TLRs in the eye, with significant implications for better understanding of ocular immunity and the pathogenesis of inflammatory eye diseases affecting the cornea, uvea, and retina. PMID- 16361680 TI - Primary nasolacrimal duct malignant mucosal melanoma. PMID- 16361679 TI - The aetiology and associations of conjunctival intraepithelial neoplasia. AB - AIM: To summarise the main causes and associations of conjunctival intraepithelial neoplasia (CIN) found in the literature to date. METHOD: Literature search using Ovid databases on the NHS Scotland E-library, Medline, and the Cochrane Library. The internet search engine Google Scholar was also used to identify relevant articles. RESULTS: The main causes and associations of CIN in the literature are sun exposure and distance from the equator; human papillomavirus infection, increased p53 expression, and HIV seropositivity. It has been found that in HIV positive individuals CIN is on the increase with people being affected with more aggressive tumours at younger ages. CONCLUSION: Given that CIN is becoming more common in HIV infected populations and that it has the potential to cause severe disability, it is important to improve our understanding of the condition. Early identification of CIN by an understanding of its aetiology and associations may enable the implementation of preventative measures to reduce its incidence and potentially lead to the diagnosis and further management of conditions such as HIV. Young people presenting with CIN in particular may benefit from HIV testing. PMID- 16361681 TI - Glycosylation type Ic disorder: idiopathic intracranial hypertension and retinal degeneration. PMID- 16361682 TI - Woodhouse Sakati syndrome associated with bilateral keratoconus. PMID- 16361683 TI - Are biometric parameters a risk factor for idiopathic macular hole formation? Results of a matched case-control series. PMID- 16361684 TI - Retinal ischaemia in type 1 neurofibromatosis. PMID- 16361685 TI - Choroiditis and exudative macular detachments in a post transplant leukaemic patient: an unusual presentation of Pneumocystis jiroveci infection. PMID- 16361686 TI - Prothrombotic and atherosclerotic risk factors lack significance in NAION patients harbouring mitochondrial DNA mutations. PMID- 16361687 TI - Inflammatory cytokine of basal and reflex tears analysed by multicytokine assay. PMID- 16361688 TI - Outcome of implementing the national services framework guidelines for diabetic retinopathy screening: results of an audit in a primary care trust. PMID- 16361689 TI - Purification of triamcinolone acetonide suspension for intravitreal injection. PMID- 16361690 TI - Late improvement in upward gaze in a patient with hydrocephalus related Parinaud dorsal midbrain syndrome. PMID- 16361691 TI - Expression of TSH-R in normal human extraocular muscles. PMID- 16361692 TI - PHEMA as a keratoprosthesis material. PMID- 16361693 TI - Trabeculotomy versus trabeculotomy-trabeculectomy for congenital glaucoma. PMID- 16361694 TI - Identification of inhibitors of the kinase activity of oncogenic V600E BRAF in an enzyme cascade high-throughput screen. AB - The Cancer Genome Project has identified several oncogenic mutations in BRAF that represent important opportunities for cancer drug discovery. The V600E BRAF mutation accounts for approximately 90% of the mutations identified. A strong case has emerged from molecular, cellular, and structural studies for the identification and development of inhibitors of this mutated BRAF protein. The authors have developed and run a high-throughput screen to find inhibitors of V600E BRAF using an enzyme cascade assay in which oncogenic BRAF activates MEK1, which in turn activates ERK2, which then phosphorylates the transcription factor ELK1. A phosphospecific antibody, Europium-labeled secondary antibody, and a time resolved fluorescent readout were used to measure phosphorylation of ELK1. Overall assay variation was 12.4%. The assay was used to screen 64,000 compounds with an overall Z' factor of 0.58 +/- 0.12. A series of 3,5, di-substituted pyridines were identified as inhibitors of the cascade assay. These compounds did not inhibit a shortened activated MEK1 to ELK1 cascade but were active (0.5-27.9 microM) in a V600E BRAF assay and represent a potential starting point for future drug discovery and development. PMID- 16361695 TI - A chemoinformatics analysis of hit lists obtained from high-throughput affinity selection screening. AB - The high-throughput affinity-selection screening platform SpeedScreen was recently reported by the Novartis Institutes for BioMedical Research as a homogeneous, label-free screening technology with mass-spectrometry readout. SpeedScreen relies on the screening of compound mixtures with various target proteins and uses fast size-exclusion chromatography to separate target-bound from unbound substances. After disintegration of the target-binder complex, the binder molecules are identified by their molecular masses using liquid chromatography/mass spectrometry. The authors report an analysis of the molecular properties of hits obtained with SpeedScreen on 26 targets screened within the past few years at Novartis using this technology. Affinity-based SpeedScreen is a robust high-throughput screening technology that does not accumulate frequent hitters or potential covalent binders. The hits are representative of the most commonly identified scaffold classes observed for known drugs. Validated SpeedScreen hits tend to be enriched on more lipophilic and larger-molecular weight compounds compared to the whole library. The potential for a reduced SpeedScreen screening set to be used in case only limited protein quantities are available is evaluated. Such a reduced compound set should also maximize the coverage of the high-performing regions of the chemical property and class spaces; chemoinformatics methods including genetic algorithms and divisive K means clustering are used for this aim. PMID- 16361696 TI - A simple, flexible, nonfluorescent system for the automated screening of neurite outgrowth. AB - Measurement of neurite outgrowth is a common assay of neurotrophic activity. However, currently available techniques for measuring neurite outgrowth are either time or resource intensive. The authors established a system in which chronic treatment of a subcloned SH-SY5Y cell line with aphidicolin and various concentrations of nerve growth factor (NGF) induced discernable alterations in proliferation and differentiation. Cells were fixed, labeled with a nonfluorescent dye, and evaluated both manually and with an automated analysis system. NGF increased multiple parameters of differentiation, including neurite length, the proportion of cells extending neurites, and branching, as well as promoting cellular survival/proliferation. Interestingly, although NGF treatment increased the total number of branches, it actually decreased the proportion of branches per neurite length. The authors observed no differences in results obtained using the manual and automated systems, but the automated system was orders of magnitude faster. To demonstrate the flexibility of the system, the authors also show that they could measure changes in differentiation induced by a small-molecule Rho kinase inhibitor, as well as by retinoic acid cotreatment with brain-derived neurotrophic factor. In addition to this flexibility, this system does not require specialized equipment or fluorescent antibodies for analysis and therefore provides a less resource-intensive alternative to fluorescence-based systems. PMID- 16361697 TI - Utilization of substrate-induced quenching for screening targets promoting NADH and NADPH consumption. AB - Oxidation of reduced nicotinamide adenine dinucleotides is a common event for many biochemical reactions. However, its exploitation for ultrahigh-throughput screening purposes is not an easy task and is affected by various drawbacks. It is known that such nucleotides induce quenching on the fluorescence of several dyes and that this quenching disappears with oxidation of the nucleotide. We have made use of this property to develop an assay for high-throughput screening with NADH and NADPH-dependent reductases. Full screening campaigns have been run with excellent assay quality parameters, and interesting hits have been identified. The method is amenable to miniaturization and allows easy identification of false positives without needing extra secondary assays. Although it is based on monitoring substrate consumption, it is demonstrated that the effect of fractional conversion on assay sensitivity is negligible. PMID- 16361698 TI - Mutated yeast heat shock transcription factor activates transcription independently of hyperphosphorylation. AB - The homotrimeric heat shock transcription factor (HSF) binds to the heat shock element of target genes and regulates transcription in response to various stresses. The Hsf1 protein of Saccharomyces cerevisiae is extensively phosphorylated upon heat shock; a modification that is under positive regulation by its C-terminal regulatory domain (CTM). Hyperphosphorylation has been implicated in gene-specific transcriptional activation. Here, we surveyed genes whose heat shock response is reduced by a CTM mutation. The CTM is indispensable for transcription via heat shock elements bound by a single Hsf1 trimer but is dispensable for transcription via heat shock elements bound by Hsf1 trimers in a cooperative manner. Intragenic mutations located within or near the wing region of the winged helix-turn-helix DNA-binding domain suppress the temperature sensitive growth phenotype associated with the CTM mutation and enable Hsf1 to activate transcription independently of hyperphosphorylation. Deletion of the wing partially restores the transcriptional defects of the unphosphorylated Hsf1. These results demonstrate a functional link between hyperphosphorylation and the wing region and suggest that this modification is involved in a conformational change of a single Hsf1 trimer to an active form. PMID- 16361699 TI - Identification of in vivo phosphorylation sites on human deoxycytidine kinase. Role of Ser-74 in the control of enzyme activity. AB - Deoxycytidine kinase (dCK) catalyzes the rate-limiting step of the deoxyribonucleoside salvage pathway in mammalian cells and plays a key role in the activation of numerous nucleoside analogues used in anti-cancer and antiviral chemotherapy. Although compelling evidence indicated that dCK activity might be regulated by phosphorylation/dephosphorylation, direct demonstration was lacking. Here we showed that dCK overexpressed in HEK 293T cells was labeled after incubating the cells with [32P]orthophosphate. Sorbitol, which was reported to decrease dCK activity, also decreased the labeling of dCK. These results indicated that dCK may exist as a phosphoprotein in vivo and that its activity can be correlated with its phosphorylation level. After purification of 32P labeled dCK, digestion by trypsin, and analysis of the radioactive peptides by tandem mass spectrometry, the following four in vivo phosphorylation sites were identified: Thr-3, Ser-11, Ser-15, and Ser-74, the latter being the major phosphorylation site. Site-directed mutagenesis and use of an anti-phospho-Ser-74 antibody demonstrated that Ser-74 phosphorylation was crucial for dCK activity in HEK 293T cells, whereas phosphorylation of other identified sites did not seem essential. Phosphorylation of Ser-74 was also detected on endogenous dCK in leukemic cells, in which the Ser-74 phosphorylation state was increased by agents that enhanced dCK activity. Our study provided direct evidence that dCK activity can be controlled by phosphorylation in intact cells and highlights the importance of Ser-74 for dCK activity. PMID- 16361700 TI - Characterization of the DNA binding and structural properties of the BRCT region of human replication factor C p140 subunit. AB - BRCT domains, present in a large number of proteins that are involved in cell cycle regulation and/or DNA replication or repair, are primarily thought to be involved in protein-protein interactions. The large (p140) subunit of replication factor C contains a sequence of approximately 100 amino acids in the N-terminal region that binds DNA and is distantly related to known BRCT domains. Here we show that residues 375-480, which include 28 amino acids N-terminal to the BRCT domain, are required for 5'-phosphorylated double-stranded DNA binding. NMR chemical shift analysis indicated that the N-terminal extension includes an alpha helix and confirmed the presence of a conserved BRCT domain. Sequence alignment of the BRCT region in the p140 subunit of replication factor C from various eukaryotes has identified very few absolutely conserved amino acid residues within the core BRCT domain, whereas none were found in sequences immediately N terminal to the BRCT domain. However, mapping of the limited number of conserved, surface-exposed residues that were found onto a homology model of the BRCT domain, revealed a clustering on one side of the molecular surface. The cluster, as well as a number of amino acids in the N-terminal alpha-helix, were mutagenized to determine the importance for DNA binding. To ensure minimal structural changes because of the introduced mutations, proteins were checked using one-dimensional (1)H NMR and CD spectroscopy. Mutation of weakly conserved residues on one face of the N-terminal alpha-helix and of residues within the cluster disrupted DNA binding, suggesting a likely binding interface on the protein. PMID- 16361701 TI - 'Compliance' is futile but is 'concordance' between rheumatology patients and health professionals attainable? PMID- 16361702 TI - Primary care guidelines in rheumatology. PMID- 16361703 TI - Vaccinate your immunocompromised patients! PMID- 16361704 TI - Effect of rituximab in refractory SLE: inhibition of Th1? PMID- 16361705 TI - Characterization of a muscle-specific enhancer in human MuSK promoter reveals the essential role of myogenin in controlling activity-dependent gene regulation. AB - Neuromuscular synaptogenesis is initiated by the release of agrin from motor neurons and the activation of the receptor tyrosine kinase, MuSK, in the postsynaptic membrane. MuSK gene expression is regulated by nerve-derived agrin and muscle activity. Agrin stimulates synapse-specific MuSK gene expression by activating GABP(alphabeta) transcription factors in endplate-associated myonuclei. In contrast, the mechanism by which muscle activity regulates MuSK gene expression is not known. We report on a 60-bp MuSK enhancer that confers promoter regulation by muscle differentiation, changes in intracellular calcium, and muscle activity. Within this enhancer, we identified a single E-box that is essential for this regulation. This E-box binds myogenin, and we showed that myogenin is necessary for not only MuSK but also nAChR gene regulation by muscle activity. Surprisingly, the same E-box functions in vivo to mediate muscle specific and differentiation-dependent gene induction in zebrafish, suggesting an evolutionary conserved mechanism of regulation of synaptic protein gene expression. PMID- 16361706 TI - Up-regulation of AMP-activated kinase by dysfunctional cystic fibrosis transmembrane conductance regulator in cystic fibrosis airway epithelial cells mitigates excessive inflammation. AB - AMP-activated kinase (AMPK) is a ubiquitous metabolic sensor that inhibits the cystic fibrosis (CF) transmembrane conductance regulator (CFTR). To determine whether CFTR reciprocally regulates AMPK function in airway epithelia and whether such regulation is involved in lung inflammation, AMPK localization, expression, and activity and cellular metabolic profiles were compared as a function of CFTR status in CF and non-CF primary human bronchial epithelial (HBE) cells. As compared with non-CF HBE cells, CF cells had greater and more diffuse AMPK staining and had greater AMPK activity than their morphologically matched non-CF counterparts. The cellular [AMP]/[ATP] ratio was higher in undifferentiated than in differentiated non-CF cells, which correlated with AMPK activity under these conditions. However, this nucleotide ratio did not predict AMPK activity in differentiating CF cells. Inhibiting channel activity in non-CF cells did not affect AMPK activity or metabolic status, but expressing functional CFTR in CF cells reduced AMPK activity without affecting cellular [AMP]/[ATP]. Therefore, lack of functional CFTR expression and not loss of channel activity in CF cells appears to up-regulate AMPK activity in CF HBE cells, presumably through non metabolic effects on upstream regulatory pathways. Compared with wild-type CFTR expressing immortalized CF bronchial epithelial (CFBE) cells, DeltaF508-CFTR expressing CFBE cells had greater AMPK activity and greater secretion of tumor necrosis factor-alpha and the interleukins IL-6 and IL-8. Further pharmacologic AMPK activation inhibited inflammatory mediator secretion in both wild type- and DeltaF508-expressing cells, suggesting that AMPK activation in CF airway cells is an adaptive response that reduces inflammation. We propose that therapies to activate AMPK in the CF airway may be beneficial in reducing excessive airway inflammation, a major cause of CF morbidity. PMID- 16361707 TI - The basic region of the diaphanous-autoregulatory domain (DAD) is required for autoregulatory interactions with the diaphanous-related formin inhibitory domain. AB - Mammalian diaphanous-related (mDia) formins act as Rho GTPase effectors during cytoskeletal remodeling. Rho binding to mDia amino-terminal GTPase-binding domains (GBDs) causes the adjacent Dia-inhibitory domain (DID) to release the carboxyl-terminal Dia-autoregulatory (DAD) domain that flanks the formin homology 2 (FH2) domain. The release of DAD allows the FH2 domain to then nucleate and elongate nonbranched actin filaments. DAD, initially discovered as a region of homology shared between a phylogenetically divergent set of formin proteins, is comprised of a core motif, MDXLLXL, and an adjacent region is comprised of numerous basic residues, typically RRKR in the mDia family. Here, we show that these specific amino acids within the basic region of DAD contribute to the binding of DID and therefore the maintenance of the mDia autoregulatory mechanism. In addition, expression of full-length versions of mDia2 containing amino acid substitutions in either the DAD core or basic regions causes profound changes in the F-actin architecture, including the formation of filopodia-like structures that rapidly elongate from the cell edge. These studies further refine our understanding of the molecular contribution of DAD to mDia control and the role of mDia2 in the assembly of membrane protrusions. PMID- 16361708 TI - Regulation of the assembly and adhesion activity of E-cadherin by nectin and afadin for the formation of adherens junctions in Madin-Darby canine kidney cells. AB - The Ca2+-independent immunoglobulin-like molecule nectin first forms cell-cell adhesion and then assembles cadherin at nectin-based cell-cell adhesion sites, resulting in the formation of adherens junctions (AJs). Afadin is a nectin- and actin filament-binding protein that connects nectin to the actin cytoskeleton. Here, we studied the roles and modes of action of nectin and afadin in the formation of AJs in cultured MDCK cells. The trans-interaction of nectin assembled E-cadherin, which associated with p120(ctn), beta-catenin, and alpha catenin, at the nectin-based cell-cell adhesion sites in an afadin-independent manner. However, the assembled E-cadherin showed weak cell-cell adhesion activity and might be the non-trans-interacting form. This assembly was mediated by the IQGAP1-dependent actin cytoskeleton, which was organized by Cdc42 and Rac small G proteins that were activated by the action of trans-interacting nectin through c Src and Rap1 small G protein in an afadin-independent manner. However, Rap1 bound to afadin, and this Rap1-afadin complex then interacted with p120(ctn) associated with non-trans-interacting E-cadherin, thereby causing the trans-interaction of E cadherin. Thus, nectin regulates the assembly and cell-cell adhesion activity of E-cadherin through afadin, nectin signaling, and p120(ctn) for the formation of AJs in Madin-Darby canine kidney cells. PMID- 16361709 TI - Regulation of protein kinase C delta by phorbol ester, endothelin-1, and platelet derived growth factor in cardiac myocytes. AB - Protein kinase C (PKC) delta is regulated allosterically by phosphatidylserine and diacylglycerol (which promote its translocation to the membrane) and by phosphorylation of Ser/Thr and Tyr residues. Although phosphorylation on Thr 505/Ser-643/Ser-662 may simply "prime" PKCdelta for activation, it could be regulatory. We examined the regulation of PKCdelta in cardiac myocytes by endothelin-1 (Gq protein-coupled receptor agonist) and platelet-derived growth factor (receptor tyrosine kinase agonist) in comparison with phorbol 12-myristate 13-acetate (PMA). All increased phosphorylation of PKCdelta(Thr-505/Ser-643) and of Tyr residues, although to differing extents. De novo phosphorylation occurred mainly after translocation of PKCdelta to the particulate fraction, and phosphorylations of Thr-505/Ser-643 versus Tyr residues were essentially independent events. Following chromatographic separation of the PKCdelta subspecies, activities were correlated with immunoreactivity profiles of total and phosphorylated forms. In unstimulated cells, approximately 25% of PKCdelta lacked phosphorylation of Thr-505/Ser-643 and displayed minimal activity (assayed in the presence of phosphatidylserine/PMA following chromatography). Endothelin-1 or PMA (10 min) promoted Thr-505/Ser-643 phosphorylation of this pool, and this was associated with an increase in total recoverable PKCdelta activity. Meanwhile, in cells exposed to endothelin-1 or PMA, the overall pool of PKCdelta translocated rapidly (30 s) to the particulate fraction and was phosphorylated on Tyr residues. This was associated with an increase in lipid-independent activity (i.e. the phosphatidylserine/PMA requirement disappeared). For endothelin-1, Tyr phosphorylation of PKCdelta and the increase in phosphatidylserine/PMA independent activity persisted after PKCdelta retrotranslocated to the soluble fraction. We concluded that, with this physiological agonist, PKCdelta becomes activated in the particulate fraction but retains activity following its retrotranslocation, presumably to phosphorylate substrates elsewhere. PMID- 16361711 TI - Epidemiology of tinea pedis and toenail tinea unguium in worshippers in the mosques in Adana, Turkey. AB - The aim of this study was to determine the prevalence of dermatomycosis and the risk factors in those who perform their prayers in the mosques in the city center of Adana Province. The diagnosis of dermatomycosis was made on the basis of direct microscopy and/or culture in addition to clinical findings. Among 461 subjects, 136 (29.5%) had tinea pedis, 23 (5.0%) had tinea unguium, and 21 (4.5%) had both infections simultaneously, with a resulting total of 180 (39.0%) cases of dermatomycosis. The causative agents by frequency were: Trichophyton rubrum (67.0%), T. mentagrophytes var. interdigitale (31.1%), and Candida albicans (1.9%). The prevalence of the foot dermatomycosis was found to be high among those who practiced ablution 3-5 times a day and did not dry their feet immediately, who used rubber shoes, and/or who shared their shoes. Pedal dermatophyte infection seems to be a major problem among the adult Muslim male population regularly attending mosques especially in 5th and 6th decades of life. PMID- 16361712 TI - There is no correlation between acne severity and AQOLS/DLQI scores. AB - The prevalence of psychiatric disorders is quite high in dermatological diseases. Psychiatric problems are also seen in acne, which involves young people in particular. In this study, carried out to evaluate the effect of acne on the quality of the social, school and private lives of university students, a total of 19 questions contained in AQOLS (Acne Quality of Life Scale) and DLQI (Dermatology Life Quality Index) were asked of the acne patients and healthy controls. The statistically significant relationships between the acne severity and AQOLS/DLQI as well as any possible differences between the AQOLS and DLQI scores in acne patients and healthy controls were investigated. Mann-Whitney U, Kruskal-Wallis and Spearman correlation tests were used in statistical analyses. For reliability, Cronbach-a analysis was used. The ages of 108 patients, 67 females (62%) and 41 males (38%), ranged between 16 and 29 (20.43+/-1.92). The ages of 100 controls, 61 females (61%) and 39 males (39%), ranged between 18 and 24 (19.45 +/- 1.35). There were no significant relationships between acne severity and AQOLS/DLQI (p=0.767). No statistically significant correlation was found between the acne patients with scars and those without scars with respect to AQOLS (p=0.253) and DLQI (p=0.255) scores. Statistically, AQOLS (p=0.000) and DLQI (p=0.000) scores of the patients with acne were found to be significantly higher as compared to the control groups. Consequently, it has been found in our study that the life quality of acne patients can be affected by reasons other than acne severity and presence of scars. The reasons can be social, personal, emotional and school-related problems of the patients. For that reason, life quality of patients should be evaluated with psychiatric measurements other than AQOLS and DLQI. In conclusion, these scores are not good enough to evaluate acne patients's quality of life. PMID- 16361710 TI - Mutational analysis of the ras converting enzyme reveals a requirement for glutamate and histidine residues. AB - The Ras converting enzyme (RCE) promotes a proteolytic activity that is required for the maturation of Ras, the yeast a-factor mating pheromone, and certain other proteins whose precursors bear a C-terminal CAAX tetrapeptide motif. Despite the physiological importance of RCE, the enzymatic mechanism of this protease remains undefined. In this study, we have evaluated the substrate specificity of RCE orthologs from yeast (Rce1p), worm, plant, and human and have determined the importance of conserved residues toward enzymatic activity. Our findings indicate that RCE orthologs have conserved substrate specificity, cleaving CVIA, CTLM, and certain other CAAX motifs, but not the CASQ motif, when these motifs are placed in the context of the yeast a-factor precursor. Our mutational studies of residues conserved between the orthologs indicate that an alanine substitution at His194 completely inactivates yeast Rce1p enzymatic activity, whereas a substitution at Glu156 or His248 results in marginal activity. We have also determined that residues Glu157, Tyr160, Phe190, and Asn252 impact the substrate selectivity of Rce1p. Computational methods predict that residues influencing Rce1p function are all near or within hydrophobic segments. Combined, our data indicate that yeast Rce1p function requires residues that are invariably conserved among an extended family of prokaryotic and eukaryotic enzymes and that these residues are likely to lie within or immediately adjacent to the transmembrane segments of this membrane-localized enzyme. PMID- 16361713 TI - Association between alopecia areata and HLA Class I and II in Turkey. AB - HLA class I and II alleles have been described in patients with alopecia areata (AA). As in other immune mediated diseases, the HLA alleles associated with AA may influence the patient's ability to respond to immune challenges from both self and non-self antigens and can offer clues to the cause, prognosis, and potential therapy for the disease. The aim of this study was to determine which HLA class I and II alleles are associated with Turkish alopecia areata patients. Sixty-three patients with AA, alopecia totalis, or alopecia universalis were included in this study and compared with seventy-six healthy transplant donors. HLA DNA typing was performed by the PCR/SSP method. The frequency of HLA-B62 was significantly higher in patients than in controls. HLA-A2, HLA-A24, HLA-B35, HLA DRB1*11, and HLA-DRB1*15 were significantly less common in patients than in the control group. PMID- 16361714 TI - An allogeneic cultured dermal substitute suitable for treating intractable skin ulcers and large skin defects prior to autologous skin grafting: three case reports. AB - Intractable skin ulcers that arise as secondary lesions from disease and full thickness skin defects that result from skin tumor excision often need autologous skin grafting to close the wound. We developed an allogeneic cultured dermal substitute (CDS) to shorten the time needed to prepare a wound bed suitable for autologous skin grafting. The CDS was prepared by plating normal human fibroblasts on a spongy matrix consisting of hyaluronic acid and atelo-collagen. The allogeneic CDS was then placed on the rinsed wound surface. This procedure was repeated twice a week for up to five weeks, until the wounds were closed by autologous skin grafting. In all three cases, after CDS treatment for two to five weeks, the wound conditions became suitable for skin grafting; these conditions had not been improved by conventional topical treatments, including topical basic fibroblast growth factor (bFGF). Healthy granulation tissue developed rapidly, concomitant with wound size reduction. The present results indicate that CDS is an excellent biological wound dressing for improving wound conditions so that they are suitable for subsequent autologous skin grafting as well as for shortening the treatment duration for skin ulcers and full-thickness skin defects. PMID- 16361715 TI - Epithelioid cell histiocytoma with underlying artery damage. AB - Although epithelioid cell histiocytoma is considered a variant of fibrous histiocytoma (dermatofibroma), the histogenesis of this condition remains controversial. A recent investigation suggested the possibility that epithelioid cell histiocytoma is an angioformative fibrous histiocytoma. We report a case of epithelioid cell histiocytoma underlying a damaged artery. This epithelioid cell histiocytoma with a central area of hemorrhage was associated with a medium-sized artery in the reticular dermis running vertically up to the lesion. Both the artery in the reticular dermis and its centrally branched artery in the subcutaneous tissue showed mural injury and intraluminal epithelioid endothelial proliferation. Immunohistochemically, the labeling rate for each of the antibodies studied in the epithlioid cells was as follows: 40% for Factor XIIIa, 5% for CD34, 20% for factor VIII-related antigen, and 10% for alpha-smooth muscle cell actin. This case may support the view that epithelioid cell histiocytoma is a vascular and angioformative fibrous histiocytoma. PMID- 16361716 TI - ITP as an initial manifestation of subacute cutaneous lupus erythematosus. AB - Subacute cutaneous lupus erythematosus (SCLE) is an entity characterized by widespread polycyclic lesions that heal without scarring. Skin lesions with marked ultraviolet sensitivity are distributed in an annular and/or psoriasiform configuration. Idiopathic thrombocytopenic purpura, which is an autoimmune disease (ITP), is mediated by a destructive immunoglobulin G antibody response to the platelets' membrane components. We report a case of subacute cutaneous lupus erythematosus initially manifested as thrombocytopenia, which was diagnosed as idiopathic thrombocytopenic purpura (ITP) and treated with splenectomy. Seven months later, development of cutaneous involvement followed the diagnosis of ITP. The clinical and histological features of the lesions were compatible with SCLE. Serological evaluations showed a negative anti-nuclear antibody test and an elevated anti-SSB/La antibody level. Symptoms for systemic involvement were negative. Although the clinical features such as photosensitivity, discoid rash, and thrombocytopenia were in favor of SLE, the patient did not fulfill the criteria of the American Rheumatism Association (ARA) for SLE. PMID- 16361717 TI - Heterotopic salivary gland tissue: a case report demonstrating evolution and association with the branchial apparatus. AB - Heterotopic salivary gland tissue (HSGT) in the lower neck is an unusual developmental anomaly with characteristic clinical and microscopic findings. The exact embryogenesis remains unclear. This rare entity must be considered in the differential diagnosis of neck mass with fistula. We present a typical HSGT totally removed using the stepladder excision technique and showing an internal fistula. Interpretation of this case from the anatomical and pathological points of view, we support the argument that the embryogenesis of HSGT is more probably related to ectodermal heteroplasia of the precervical sinus of His and further conclude that an association with branchial cleft sinus may exist and cannot be seen as an exclusion criteria for diagnosis of HSGT. Due to possible but infrequent neoplastic transformation, it is important to check HSGT in every encountered cervical anomaly related to any branchial apparatus derived lesion. PMID- 16361718 TI - A case of Werner syndrome with three primary lesions of malignant melanoma. AB - Three primary lesions of malignant melanoma developed in a 44-year-old Japanese woman with Werner syndrome. One lesion was on the right large pudental lip and the others in distinct locations on her left sole. After the wide local excision of these tumors, the wound of the large pudental lip was sutured, and the defects on the sole were covered with skin grafts. After one course of chemotherapy consisting of dacarbazine, nimustine, vincristine sulfate and local injection of Interferon beta were performed, severe myelosupression occurred and continued for two months. Defective production of WRN protein was confirmed by Western blotting, although the three representative mutations in Japanese patients, mutations 1, 4 and 6, which include over 90% of the Japanese patients, were not detected. We also reviewed 26 cases of malignant melanoma associated with Werner syndrome (WS), including ours. PMID- 16361719 TI - Primary varicella infection associated with stevens-johnson syndrome in a Turkish child. AB - Stevens-Johnson syndrome (SJS) is defined as a severe erythema-multiforme-like eruption of the skin and lesions of the oral, genital and anal mucosa and hemorrhagic crusting on the lips; it is associated with fever, headache, and arthralgia. The disease is a part of a continuum of immunologically mediated mucocutaneous diseases at various grades of severity. SJS is often induced by drugs, but the pathophysiologic mechanism is completely unknown. Patients and their first degree-relatives may have genetic defects in their metabolic pathways that lead to the accumulation of toxic metabolites. However, viral infections are known triggers of this skin disorder. Varicella zoster virus has been only very rarely reported as an etiological agent in SJS, despite its high incidence as a pathogen in childhood. In this study, we report a case of primary varicella infection associated with SJS because of its rare presentation. To the best of our knowledge, our case is the youngest child in the literature. PMID- 16361720 TI - Lipoid proteinosis: an unusual presentation with verruca vulgaris. AB - Lipoid proteinosis (LP) is a very rare, autosomally recessive, inherited disease. It is also known as Urbach Wiethe disease or Hyalinosis cutis et mucosa. LP usually runs a benign, stable, and slowly progressing course. We present one LP case with additional unusual dermatologic involvement; verruca vulgaris was superimposed on papular lesions of LP. To our knowledge, this case is the first and only one described in the literature. In this presentation, we discuss the possibility of predisposition to Human Papilloma Virus (HPV) infections in LP. PMID- 16361721 TI - A case of linear lichen planus. AB - Linear lichen planus (LLP) is a very rare form of lichen planus (LP) characterized by a linear distribution of the lichenoid lesions. It usually follows the lines of Blaschko with unilateral involvement. We report such a case of LLP. A 50-year-old man presented with slightly pruritic, linear, violaceous, papular lesions on the right side of his jaw. A biopsy specimen demonstrated the typical histology of LP. Based on the clinical and pathological findings, the diagnosis was LLP. PMID- 16361722 TI - Drug-induced linear IgA bullous dermatosis. AB - We report the case of a 69-year-old Japanese woman with multiple blistering lesions covering almost her whole body. Linear IgA and C3 depositions were seen at the basement membrane zone on direct immunofluorescence (IF). Linear IgA bullous dermatosis (LABD) is one of the autoimmune diseases resulting in subepidermal blisters. It is clinically similar to bullous pemphigoid and IF is required to distinguish the two diseases. In this case, the blistering lesions appeared after vancomycin treatment. This drug was strongly suspected as a cause of LABD in light of the clinical course of the patient even though a drug lymphocyte stimulating test was negative. Among the various implicated causative drugs, vancomycin is the most commonly associated with LABD. PMID- 16361723 TI - Friction melanosis and striae distensa caused by stretch training on a bench press. PMID- 16361724 TI - Prurigo pigmentosa on a patient with soft-drink ketosis. PMID- 16361725 TI - Evaluation of psoriatic patients with HCV infection. PMID- 16361726 TI - Various patterns of repigmentation after narrowband UVB monotherapy in patients with vitiligo. PMID- 16361727 TI - The various effects of four H1-antagonists on serum substance P levels in patients with atopic dermatitis. AB - Antiallergic drugs have various actions against allergy-associated cells and molecules as well as antihistamic properties. We studied the effects of antiallergics on the serum levels of substance P. Patients with atopic dermatitis were treated with one of four oral H1-antagonists for 14 days, and the serum level of substance P was measured before and after treatment in parallel with several atopic severity markers. Olopatadine significantly decreased the substance P level. This is in accordance with its known downmodulatory effect on tachykinin release. In contrast, cetiridine and fexofenadine unexpectedly increased the substance P level. In patients administered cetiridine, the blood severity markers for atopic dermatitis, including lactate dehydrogenase, eosinophil number, and the soluble forms of IL-2R, E-selectin, VCAM-1 and ICAM-1 were reduced after the treatment. Therefore, the elevation of SP was unrelated to the deterioration of atopic dermatitis but rather associated with improvement. Our study suggests that antiallergics can be divided into substance P-increasing and -decreasing types and raises the possibility that the increment of substance P by the former type is caused by the competitive occupation of substance P receptors. PMID- 16361728 TI - Bleaching phototrichogram: an improved method for hair growth assessment. AB - The phototrichogram (PT) is a non-invasive tool for monitoring hair growth in vivo. However, the majority of PT techniques are of little use to clinicians bacause they are time-consuming and/or difficult to perform. Consequently, there is a need for an easy and time-saving PT technique. This study was performed to evaluate the accuracy and availability of a modified PT, bleaching phototrichogram (BPT), by comparing it with the conventional phototrichogram (CPT). Ten healthy male volunteers (age range 26-33 years) comprised the study subjects. Hairs were clipped from two adjacent circular areas in the occipital region of each subject. The clipped hairs in the first area were bleached and photographed two days later. The second area was photographed just after clipping and two days later. In each area, the following biological parameters of hair growth were analyzed; 1: hair density (number/cm2), 2: anagen hair ratio (%), 3: linear hair growth rate (mm/day), 4: non-vellus hair ratio (%) and 5: anagen hair thickness (microm). The hair density, anagen hair ratio, linear hair growth rate, and anagen hair thickness by BPT were similar to the CPT values with no significant differences, although the non-vellus hair ratio by BPT was higher than the CPT value. The BPT has a higher clinical usefulness than the CPT because it saves much time with satisfactory accuracy. PMID- 16361729 TI - Topical antibiotic treatment of impetigo with tetracycline. AB - Forty-nine children aged 0.2-13 years with bullous and eroded lesions, from which Staphylococcus aureus was isolated, were diagnosed with impetigo and entered into a randomized, open-labeled trial of topical oxytetracycline hydrochloride (tetracycline) compared with a combination of topical tetracycline and oral antibiotics. After one week of topical tetracycline treatment, 22 of the 28 patients were clinically cured, and the remaining six patients had improved. In the other treatment group, 14 patients of 21 were clinically cured and 7 patients improved by the combination of topical tetracycline and oral antibiotics. There were no significant differences between the two groups. Therefore, the present study suggests that topical tetracycline treatment is effective for the treatment of impetigo. PMID- 16361730 TI - Narrow-band UVB treatment of vitiligo in Chinese. AB - Narrow-band ultraviolet B (NBUVB) phototherapy has recently been reported to be an effective and safe treatment modality for vitiligo. In the present report, we evaluated the efficacy and safety of NBUVB therapy for vitiligo in Chinese patients. Seventy-two vitiligo patients treated from 2000 to 2003, were included retrospectively (male: female=33:39, mean age: 38.5). Among them, 61 were non segmental type and 11 the segmental type. Treatments were given two to three times a week for a maximum period of one year with an initial dose of 0.2 J/cm2 and a 0-20% increment each session (mean accumulation dose: 155.3 J/cm2). Computer image analysis by Supervise classification was used to estimate the area of vitiligo involvement before and after treatment. An excellent response (75 100% area of repigmentation) was obtained in 9 patients (12.5%) and a good response (50-75%) in 24 (33.3%), a moderate response (25-50%) in 20 (27.8%), and a poor response (0-25%) in 19 (26.4%). In summary, 45.8% of our patients had more than 50% repigmentation. Burns were a side effect in 5 patients (7%) and transient erythema with itching or xerosis was noted in 5 patients (7%). These results indicate that NBUVB phototherapy is an effective and safe treatment choice for generalized vitiligo. PMID- 16361731 TI - A case of bullous congenital ichthyosiform erythroderma (BCIE) caused by a mutation in the 1A helix initiation motif of keratin 1. AB - Bullous congenital ichthyosiform erythroderma (BCIE) is an autosomally dominant inherited disorder characterized by erythematous, erosive, and bullous skin lesions over the entire body at birth and abnormal hyperkeratosis on the palmoplantar sufaces as the patient grows older. BCIE is caused by a mutation in the keratin 1 (K1) and/or keratin 10 (K10) genes, and most pathogenic mutations are found within the helix initiation and termination motifs of the central helical rod domain (K1 and K10) or the upstream H1 homology domain (K10). In addition to inherited cases, sporadic cases due to a new mutation account for approximately half the total cases of BCIE. We report herein a typical sporadic case of BCIE with erythroderma, erosion, and blisters on the entire body surface at birth and palmoplantar and flexuaral areas of hyperkeratosis in the later stage. We found in this case a novel mutation, 559C to T, at amino acid position 187, which resulted in a leucine to phenylalanine substitution within the helix initiation motif of K1. PMID- 16361732 TI - Childhood bullous pemphigoid successfully treated with diaminodiphenyl sulfone. AB - Bullous pemphigoid (BP) is an acquired autoimmune blistering disease which predominantly affects the elderly. It is rare in children and infants. We reported a 14-year-old girl presenting with a month history of relapsing tense bullae on the face and extremities. Histopathological examination of the lesional skin revealed a subepidermal bulla with infiltration of eosinophils, neutrophils, and lymphocytes. Direct immunofluorescence showed linear deposits of IgM and C3 at the basement membrane zone. Indirect immunofluorescence using normal human skin sections as a substrate detected IgG anti-basement membrane zone antibodies in the patient's serum and that using 1M NaCl split skin sections showed that the patient's antibodies bound to the epidermal side of the split skin. Immunoblot analysis using normal human epidermal extracts demonstrated the presence of autoantibodies against the 230-kDa BP antigen. Furthermore, the patient's serum reacted with the recombinant protein of the NC16a domain of the 180-kDa BP antigen by immunoblot analysis and enzyme-linked immunosorbent assay. Our patient showed significant improvement of the skin lesions with systemic administration of diaminodiphenyl sulfone. PMID- 16361733 TI - Spindle cell squamous cell carcinoma showing continuous mesenchymal dedifferentiation in a single tumor. AB - We report the case of an 85-year-old man who presented with an enlarging tumor on the right temple. Histologically, spindle-shaped cells were proliferating continuously within well-differentiated squamous cell carcinoma that was immunohistochemically negative for vimentin and alpha-smooth muscle actin (ASMA) but positive for cytokeratin (AE1/3). These spindle-shaped cells expressed vimentin and ASMA but not cytokeratin (AE1/3). Intermediate cells with round nuclei and small amounts of cytoplasm displayed slight expression of vimentin, ASMA, and cytokeratin. This case illustrated spindle cell squamous cell carcinoma with mesenchymal metaplasia of malignant epithelial cells. PMID- 16361734 TI - Two cases of telangiectasia macularis eruptiva perstans demonstrated by immunohistochemistry for c-kit (CD 117). AB - Telangiectasia macularis eruptiva perstans (TMEP) is an uncommon form of cutaneous mastocytosis that occurs exclusively in adults. Histologically, TMEP presents with scattered mast cells lined up around the dilated capillaries and venules of the superficial vascular plexus. In some cases, the number of mast cells falls within the range observed in normal skin and therefore cannot be detected by routine histologic examination. We used immunohistochemical staining for c-kit (CD 117) for the definitive diagnosis in two patients with TMEP. One of them was successfully treated with topical application of pimecrolimus. PMID- 16361735 TI - A case of epidermotropic metastatic malignant melanoma with multiple nodular lesions of the scalp. AB - Epidermotropic metastatic malignant melanoma (EMMM) is a form of metastatic malignant melanoma that has dermal cell nests with epidermotropism and specific histopathological features. We report a patient with eight nodular lesions of the scalp with histopathological findings compatible with EMMM. The tumors developed one year before consultation and increased in size simultaneously. The histopathological findings of all eight tumors were very similar. The tumor cells were located mainly in the dermis and partly in the basal layer of the epidermis. They contained melanin pigments and were positive for anti-HMB45 antibody. The tumors did not respond to combination chemotherapy with dacarbazine, nimustine, vincristine, and interferon-beta. Therefore, all the tumors were surgically removed. No local relapse, distant metastasis or re-elevation of plasma 5-S cysteinyldopa was identified during nine months of follow-up. Histopathologically, all eight tumors lacked apparent vascular invasion, which may be related to a slow clinical course of the present case. PMID- 16361736 TI - Pyoderma gangrenosum in association with juvenile rheumatoid arthritis. AB - A 17-year-old girl presented with multiple, painful, erythematous blisters and ulcerated lesions on the shins and buttocks. She also had arthralgia. She had suffered from juvenile rheumatoid arthritis (JRA) and received anti-inflammatory agents and oral glucocorticoids for eight years. A biopsy of a lesion showed epidermal ulceration with marked neutrophilic infiltrates in the dermis. The patient was diagnosed with pyoderma gangrenosum (PG). PG is an uncommon cutaneous ulceration within the spectrum of the neutrophilic dermatoses that is reported in association with a number of systemic disorders, including inflammatory bowel disease, hematologic disease, internal malignancies, arthritis, immune abnormalities, and solid tumors. To our knowledge, this is the first reported case of PG associated with JRA. PMID- 16361737 TI - A case of vulvitis granulomatosa. AB - Merkersson-Rosenthal syndrome (MRS) is characterized by cheilitis or pareitis granulomatosa, facial palsy, and fissured tongue. Sometimes the forehead, eyelids and chin are involved, and occasionally genital lesions are present. A vulval lesion of MRS is named vulvitis granulomatosa. Tranilast has been described as an effective treatment for cases of MRS, granulomatous cheilitis, and granulomatous blepharitis. Herein we report a successful treatment with tranilast of an adult female patient with vulvitis granulomatosa. PMID- 16361738 TI - Keloid formation on an inconspicuous penis. AB - Inconspicuous penis is an umbrella term used to categorize several anatomic anomalies that occur in newborn boys. The consequences of this anomaly may be catastrophic unless it is accurately diagnosed. Repeated circumcisions will accentuate the problem without any solution. Keloid formation on the penis is exceptionally rare even though circumcision is a world-wide procedure. Herein, we present a 13-year-old boy with a buried glans penis, micturation difficulty, and keloid formation. He was previously circumcised twice before admission to our clinic. A surgical correction together with intralesional steroid injection was performed. The outcome of this procedure was satisfactory. We conclude that repeated circumcision may not be the best choice of treatment for keloid formation on the penis. PMID- 16361739 TI - Relapsing Henoch-Schonlein purpura in an adult patient associated with hepatitis B virus infection. AB - Henoch-Scholein purpura is usually a disease of children presenting with arthralgia, abdominal pain, renal involvement, and palpable purpura. Viral and bacterial infections may have a role in its etiology. We present a 32-year-old male patient with recurrent Henoch-Scholein purpura in association with a chronic hepatitis B infection of ten years duration. The patient had received lamuvidine and interferon-alpha for the treatment of hepatitis B infection for a year. The skin lesions disappeared with the treatment of the hepatitis B infection. Four months after discontinuation of the therapy, the purpuric papules reappeared with reactivation of the hepatitis B infection. Although rarely reported, hepatitis B virus infection should be considered in patients with Henoch-Scholein purpura. PMID- 16361740 TI - A case of photocontact urticaria induced by photodynamic therapy with topical 5 aminolaevulinic acid. AB - Photodynamic therapy (PDT) with topical application of 5-aminolaevulinic acid (ALA) is a promising new treatment option for the management of various cutaneous malignancies. Generally, topical ALA-based PDT has relatively insignificant adverse effects of transient character; these include itching, stinging or burning pain and slight to moderate erythema. We describe the first case of photocontact urticaria induced by topical ALA-based PDT for the treatment of unilesional mycosis fungoides. Although the first treatment session resulted merely in mild erythema, the second PDT caused marked urticaria corresponding to the PDT-applied area with an intolerable stinging sensation. A photopatch test demonstrated that black light and visible light irradiation after topical ALA provoked an urticarial reaction in the patient's uninvolved skin. These observations suggested an allergic pathogenesis for the wheal formation induced by PDT with topical ALA in this case. Photocontact urticaria should be considered as a possible adverse effect in ALA-based PDT. PMID- 16361741 TI - Atypical fibroxanthoma on a bald scalp. AB - We present the clinical, histopathological and immunohistochemical findings of an atypical fibroxanthoma (AFX) on the bald scalp of an 81-year-old French man who had worked at a private high school in Japan as a janitor for over 40 years. The patient had a history of basal cell carcinoma on the nape, and chronic solar radiation seemed to be a predisposing factor in the pathogenesis of this association. This case showed the typical clinical and histopathological characteristics of AFX, and the immunohistochemical results suggested differentiation of histiocytes and myofibroblasts. The AFX was completely resected, and the patient has not had tumor recurrence or metastasis for over four postoperative years. This case therefore provides further support to the theory that AFX displays a clinically benign course, even though it is essentially a malignant tumor histologically located in the dermis. Therefore, we must excise AFX completely with great care and perform regular physical examinations for several years after operation. PMID- 16361742 TI - A case of cutaneous odontogenic sinus. AB - Despite the fact that cutaneous sinus tracts of odontogenic origin are well documented, the condition is still commonly misdiagnosed, because chronic periapical periodontitis may be asymptomatic and is rarely open to the skin. A 75 year-old Japanese woman presented to our clinic with the chief complaint of a left cheek skin lesion with mild pain. Physical examination revealed a subcutaneous nodule covered with erythematous skin on her left buccal region. Cultures from the subcutaneous nodule grew Bacteroides species and Peptostreptococcus micros but did not yield acid-fast bacilli, fungi, or Actinomyces. Stains of smeared pus showed a considerable number of Gram-negative rods. The histopathological examination revealed a focal abscess formation in the lower dermis and subcutaneous tissue. Dental evaluation, including an orthopantogram, showed a radiolucent alveolar area at the left lower first molar apex, suggesting a periapical abscess. Antibiotic therapy for three weeks associated with surgical root canal therapy eliminated the subcutaneous nodule. A high degree of suspicion is required to correctly diagnose a lower facial lesion as being of odontogenic origin, and prompt dental evaluation should be considered. PMID- 16361743 TI - Discrete papular mucinosis on the back. AB - A rare case of papular mucinosis on the back of a 78-year old man is described. The patient developed approximately 20 discrete papules on the upper back. His serum glucose concentration was borderline elevated, but no other systemic symptoms or signs were apparent. A biopsy revealed mucin deposits only in the upper reticular dermis. The papules were distinctly different from lichen myxedematosus in clinical and histochemical appearance. The present case may represent very early-stage lichen myxedematosus complicated by borderline diabetes mellitus. PMID- 16361744 TI - Diagnosis of epidermolytic palmoplantar keratoderma in a very early stage by gene analysis. PMID- 16361745 TI - Severe pemphigus vulgaris treated with rituximab (Mabthera). PMID- 16361746 TI - Effects of calcipotriol cream and ointment, clobetasol cream and ointment and tretinoin cream on the erythemogenicity of UVB. AB - Various studies have shown the blocking effects of topical agents on UVB penetration, which can be used in combination with phototherapy. In this study, the photoprotective effects of 0.005% calcipotriol, 0.05% clobetasol-17 propionate, and 0.1% tretinoin, which can be used in combination with broad-band UVB, were investigated in an in vivo test. In a study group of 20 patients, phototests were performed to determine minimal erythema doses (MED) and the tests were repeated with thin (0.1 cc/25 cm2) and thick (0.3 cc/25 cm2) calcipotriol, clobetasol-17-propionate, and tretinoin in cream forms and sunscreen. After determining the MED, the test was repeated in another 20 patients with thin and thick calcipotriol and clobetasol-17-propionate in both cream and ointment forms and sunscreen. MED was increased with thin and thick applications of all agents. Moreover, the photoprotective effects of each agent increased with their thick applications compared with thin ones. The application of calcipotriol cream and ointment, clobetasol cream and ointment, and tretinoin cream, all of which can block UVB, is not recommended just before phototherapy. PMID- 16361747 TI - Skin cancer screening on a fishing island and in an inland agricultural area of Japan. AB - We performed skin cancer screening from 2000 to 2004 at two locations in Japan's Oita Prefecture: Himeshima, a small fishing island, and Naoiri, an inland agricultural area. We found 108 and 21 cases of AK in Himeshima and Naoiri, respectively. None of the AKs transformed into SCC, and 21.7% of the AKs underwent spontaneous remission during our observation period. The prevalence and incidence of AK in Himeshima were five times higher than in Naoiri: 1,399 and 826 per 100,000 population, respectively, in the fishing village, vs. 261 and 164 in the agricultural community. Seven and three cases of BCC were observed in Himeshima and Naoiri, respectively. There were two cases of SCC in Himeshima. The highest risk ratio of skin types I to III was 9.2 in Himeshima. Although people engaged in outdoor occupations are thought to be more prone to skin cancer and precancerous skin lesions, our results suggested different potentials for AK in people engaged in different outdoor occupations. PMID- 16361748 TI - Dapsone hypersensitivity syndrome: a clinico-epidemiological review. AB - Diaminodiphenyl sulphone (dapsone) is a drug of choice in the treatment of leprosy. It is also useful for the treatment of many neutrophilic and other dermatoses. Dapsone hypersensitivity syndrome is a rare but well recognized serious adverse effect characterized by fever, skin rashes, generalized lymphadenopathy, hepatitis, and hepato-splenomegaly. Twenty-six patients with dapsone hypersensitivity syndrome were studied for clinical profile, outcome, and prognosis. The male:female ratio was 2.2:1, and the mean age was 33.19 years (range 13 to 64 years). The interval between start of dapsone therapy and appearance of symptoms varied from 2-7 weeks (mean 29.82 days). Twenty-four patients received dapsone as a part of multi-drug therapy for leprosy; the other two patients received dapsone for lichen planus and acne vulgaris. Exfoliative dermatitis was the most common cutaneous manifestation followed by erythematous maculo-papular eruption and Stevens-Johnson syndrome-like lesion. The other common systemic manifestations were: fever (26 cases), itching (22 cases), lymphadenopathy (21 cases), jaundice (21 cases), pallor (20 cases), hepatomegaly (19 cases), and pedal edema (14 cases). Investigation profile revealed elevated levels of serum liver enzymes in 100% of patients, elevated erythrocyte sedimentation rate in 92.3%, raised bilirubin in 84.6%, leucocytosis in 69.23%, low hemoglobin (<9 gm/dl) in 46.15% and hypoproteinemia in 42.3%. Eosinophilia, hemolytic anemia, and reticulocytosis count were found in 4 patients each. All the patients had favorable outcomes except three who died due to hepatic failure. Medical personnel must be aware of this potentially fatal syndrome, because it can cause considerable morbidity and mortality. PMID- 16361749 TI - Inflammatory stage of disseminated superficial porokeratosis. AB - Disseminated superficial porokeratosis (DSP) is a keratinization disorder characterized by multiple small lesions with a slightly elevated, sharply defined ridge over the whole body. Unusual DSP cases with acute exacerbation of their lesions accompanied by severe pruritus have been reported and designated as "eruptive pruritic papular porokeratosis" or "inflammatory DSP". Histologically, the pruritic lesions in the majority of these unusual DSP cases had a dense infiltration of eosinophils and lymphocytes in the vicinity of blood vessels in the upper dermis. In this report, we describe an additional case of DSP with a similar clinical course and histopathological findings. A review of the literature showed that the pruritic condition in these unusual DSP cases can be transient and is not necessarily related to spontaneous regression. We propose the term "Inflammatory stage of DSP" for describing this unusual variant of DSP. PMID- 16361750 TI - A case of giant porokeratosis with vestiges of a cornoid lamella. AB - We report a case of giant porokeratosis combined with ulcerative squamous cell carcinoma. In our patient, we biopsied the skin, including the edge of the skin lesion, four times until we obtained histologic proof of a cornoid lamella. After we had established the diagnosis, we totally excised the affected skin including a 10-mm safety margin, because such lesions tend to develop into skin cancer. We reconstructed the excised area using a skin graft. After the operation, we took 13 samples of skin (seven from the lower leg and six from the sole of the foot) from the edge of the main lesion, including a putative cornoid lamella. Five of the six samples from the sole and one of the seven from the lower leg were demonstrated histologically to include a cornoid lamella. Our results suggest that skin biopsies should be taken from various sites at the edge of a giant porokeratotic lesion and in particular from the prominent ridge to prove the presence of a cornoid lamella. Shallow keratin-filled invaginations and underlying squamous cells with eosinophilic cytoplasm were observed at the edge of the lesion on the lower thigh, which might suggest a diagnosis of porokeratosis with an incomplete cornoid lamella if porokeratosis was strongly suspected from the patient's clinical features. Better recognition of giant porokeratosis is required, so that an earlier diagnosis can be made and appropriate therapy initiated in a timely manner. PMID- 16361751 TI - Possible spontaneous regression of a metastatic lesion of keratoacanthoma-like squamous cell carcinoma in a regional lymph node. AB - We report herein a case of keratoacanthoma-like squamous cell carcinoma in an 82 year-old male that showed regional lymph node metastasis followed by self regression of the metastatic lesion. This case is consistent with the previously reported observation that tumors compatible with the clinical features of keratoacanthoma may have malignant biological potential, however, it also demonstrates that even metastatic lesions may exhibit self-regression similar to that observed for primary lesions. Despite the view that keratoacanthoma-like squamous cell carcinoma may represent a variant of squamous cell carcinoma, occasionally it undergoes a unique clinical course. PMID- 16361752 TI - Epithelioid sarcoma associated with lung adenocarcinoma. AB - Epithelioid sarcoma is an uncommon soft tissue malignancy which is often misdiagnosed as necrobiotic granuloma or chronic inflammation. Many patients have local recurrence and distant metastasis because of its infiltrating growth. We report a 49-year-old Japanese man who had an ulcer with surrounding erythema on the left forearm after he sustained a bruise on this spot. His mother and father had died of rectal and bladder carcinoma, respectively, and he had also had a seminoma previously. Therefore, there was familial accumulation of malignancies. A skin biopsy revealed polygonal, plump, or spindle-shaped epithelioid cells and lymphocytes infiltrating through the dermis. Immunohistochemical studies showed positive staining for cytokeratins, epithelial membrane antigen, CD34 and vimentin and negative staining for desmin, CD68, HHF-35, 1A4 and p53. These clinical and histological features were diagnostic of epithelioid sarcoma. CT scans detected nodules in the right lung. A lung biopsy revealed that he had well differentiated adenocarcinoma which expressed p53. Mutations in the TP53 gene were not searched for. We selected conservative treatment because the patient did not want surgical wide resection or amputation. Therefore, radiation, thermotherapy, application of 0.5% doxorubicin hydrochloride ointment and right lower lobectomy with lymph node dissection were performed. The size of the ulcer and the tumor invasion along the fasciae of muscles were decreased, however, metastasis of the epithelioid sarcoma was detected in the lymph node of the left axilla four years after the diagnosis. PMID- 16361753 TI - Cardio-facio-cutaneous syndrome: two cases in the same generation. AB - A 4-year-old Japanese boy, the youngest of three brothers, presented with ichthyosiform hyperkeratosis over his whole body, eczematous erythema with partial desquamation and erosion on the flexor side of the joints of extremities, the fossa axillaries, and the genital and buttock regions, and total hair loss on the scalp and the absence of eyebrows and eyelashes. In addition to the ichthyotic eruptions and hair abnormalities, he also had a ventricular septal defect, mental retardation, growth retardation, characteristic facial features such as a depressed nasal bridge, low-set ears, and ocular hypertelorism; therefore, he was diagnosed with cardio-facio-cutaneous (CFC) syndrome. The patient's family did not have a history of consanguineous marriage. The parents and the eldest son were healthy. However, the second son, also born with ichthyosiform hyperkeratosis over his whole body, total hair loss on the scalp, myocardial deficiency, mental retardation, growth retardation, and characteristic facial features, had died of pneumonia and sepsis at the age of 1.5 years. Because the middle brother had the same disease, the present case is considered to be a rare case of CFC syndrome with in a single generation. PMID- 16361754 TI - Unilateral Beau's lines associatd with a fingertip crushing injury. AB - Beau's lines are transverse depressions or ridgings of the nail surface that are the result of the temporary arrest of nail matrix formation. They are caused by systemic diseases, local trauma, drugs, and infection. Generally, Beau's lines after a fingertip injury occur on only the affected finger, not on the unilateral extremity. In this report, we present the case of a 36-year-old man who developed transverse ridgings in the nail plates of all five fingers of the affected extremity after a fingertip (thumb) crushing injury. During the follow up, the growth rate of affected fingernails was normal. The transverse ridgings advanced with the linear growth of the nails and eventually disappeared. To the best of our knowledge, no association between unilateral Beau's lines and fingertip injury has been reported in the English literature. PMID- 16361755 TI - Thrombocytopenia possibly induced by etretinate in a psoriatic patient. AB - A patient with psoriasis vulgaris developed multiple purpura on the extremities and hemorrhage at the oral mucosa and gingiva with marked thrombocytopenia. This thrombocytopenia was possibly induced by a retinoid agent, etretinate, from the clinical course and data. The total amount of etretinate administered was 2,410 mg over 191 days (41.9 mg/kg body weight, average 15.3 mg/day). A platelet transfusion partially restored the platelet count and the purpura and gingival hemorrhage disappeared approximately 10 days after the cessation of etretinate. However, the platelet count remains at 60-80 x 103/mm3 after two and a half years without etretinate therapy. Although there are only a few case reports of etretinate-induced thrombocytopenia, we should pay more attention to the peripheral platelet count during etretinate therapy. PMID- 16361756 TI - A case of bleomycin-induced acral erythema (AE) with eccrine squamous syringometaplasia (ESS) and summary of reports of AE with ESS in the literature. AB - Chemotherapy-induced acral erythema (AE) is primarily induced by hydroxyurea, methotrexate, and cytarabine, although there are rare reports of AE induced by combination chemotherapy containing bleomycin. It is thought that the accumulation of chemotherapeutic drugs in eccrine glands may cause eccrine squamous syringometaplasia (ESS), which is characterized by metaplasia and focal necrosis of the epithelium of the eccrine duct. ESS is occasionally detected in conjunction with AE, but such occurrences are relatively uncommon. This is the first report of AE with ESS induced by the administration of bleomycin alone. We also provide a summary of past cases of AE with ESS in the literature. PMID- 16361757 TI - Azithromycin in the treatment of papulopustular rosacea. PMID- 16361758 TI - A case of acute generalized exanthematous pustulosis caused by lincomycin. PMID- 16361759 TI - A Japanese case of nuchal-type fibroma. PMID- 16361760 TI - Sequential development of herpes zoster duplex unilateralis during oral famciclovir treatment. PMID- 16361761 TI - Regional variations in hepatic vascular occlusion. PMID- 16361762 TI - Helicobacter pylori-induced apoptosis in pathogenesis of gastric carcinoma. AB - BACKGROUND: Despite a possible role of Helicobacter pylori in gastric carcinoma (GC), its pathogenesis is not clear. There is scanty data on apoptosis in GC in relation to H. pylori and CagA antibody. Therefore, we studied gastric epithelial apoptosis in GC and non-ulcer dyspepsia (NUD) with or without H. pylori infection, and the degree of apoptosis in relation to CagA antibody status. METHODS: 20 patients each with GC and NUD were investigated for H. pylori using rapid urease test (RUT), IgG anti-H. pylori and anti-CagA antibodies, histology of endoscopically normal-looking mucosa for H. pylori, intestinal metaplasia (IM), and apoptosis using TUNEL assay. Positivity to one tissue-based (RUT or histology) and one serology based (anti-H. pylori or CagA IgG) test was taken as diagnostic of active H. pylori infection, and negative result in both tissue based tests suggested its absence. RESULTS: Patients with GC more often had anti H. pylori IgG (16 of 20 vs. 8 of 20; p=0.02) and a trend towards higher apoptotic index (AI) (48.6 [19.2 to 71.7] vs. 41.4 [11.7 to 63.6]; p=0.06) than NUD. AI was higher in GC (66.7 [57.5 to 71.7] vs. 32.6 [19.2 to 39.8]; p<0.0001) and NUD (58.6 [50.7 to 63.6] vs. 24.4 [11.7 to 32.2]; p<0.0001) infected with H. pylori than in those without infection. AI was also higher in GC than in NUD with H. pylori infection (66.7 [57.5 to 71.7] vs. 58.6 [50.7 to 63.6]; p=0.01). Four of the 20 patients with GC and none with NUD had IM (p=ns). There was no difference in AI in relation to CagA antibody. AI positively correlated with patients' age in presence of H. pylori infection (correlation coefficient=0.5, p=0.03) but not in its absence. CONCLUSION: Exaggerated apoptosis may play a role in H. pylori mediated gastric diseases including carcinogenesis. AI increases with aging in patients infected with H. pylori. PMID- 16361763 TI - Evaluation of Mannheim peritonitis index and multiple organ failure score in patients with peritonitis. AB - BACKGROUND: Early classification of severity of peritonitis by scoring systems, including the Mannheim peritonitis index (MPI) and the multiple organ failure (MOF) score, modulates surgical and medical management. AIM: To predict outcome of patients with peritonitis using the MPI and MOF scoring systems. METHODS: Prospective evaluation of the MPI and MOF score was performed in 80 consecutive patients with peritonitis who underwent uniform surgical treatment. Risk ratios were calculated for the MPI and other patient characteristics. Risk ratio was not calculable for the MOF score. RESULTS: Overall in-hospital mortality rate was 17.5%, including 80% of patients with MPI>29. In non-survivors the mean score was 4.8 (SD 1.46) and 33.07 (4.81) for the MOF score and MPI, respectively. Survivors had mean MOF score of 0.28 (0.20) and mean MPI of 19.39 (6.68). CONCLUSION: The MPI and MOF score provide simple and objective means to predict the outcome of patients with peritonitis. PMID- 16361764 TI - Organ-preserving multimodality management of squamous cell carcinoma of anal canal. AB - AIM: To study the efficacy of an organ-preserving, sequential chemoradiation therapy for squamous cell carcinoma of the anal canal, and of salvage surgery in those in whom this treatment fails. METHODS: Forty biopsy-proven untreated patients (28 men) with squamous cell carcinoma of the anal canal received two cycles of chemotherapy using cisplatin and methotrexate, followed by 45 to 60 (median 50) Gy external beam radiotherapy. Salvage surgery was offered to those in whom this treatment failed. Overall survival, disease-free survival and colostomy-free survival were analyzed. RESULTS: Most patients (n=35; 87%) had T3 or T4 lesions and 5 (12.5%) had involvement of inguinal nodes. Thirty-one patients (77.5%) had complete response after chemoradiation. Only three patients (7.5%) developed chemotherapy-related grade 3 mucositis and myelosuppression. Radiotherapy-related toxicity included grade III cystitis in one patient and grade III proctitis in three patients. Three patients had post-treatment anal stenosis requiring repeated dilatation and two had chronic non-healing ulcers at the anal verge. Nine patients had failure of chemoradiation or disease recurrence; of these, only 5 could undergo salvage surgery. After a median follow up of 60 months, overall survival, disease-free survival and colostomy-free survival were 80%, 77.5% and 72.5%, respectively. CONCLUSION: Chemoradiation is effective in the treatment of squamous cell anal cancer and has acceptable toxicity. Surgical salvage may be useful in those with failure of this treatment. PMID- 16361765 TI - Thrombophilic conditions in non-cirrhotic portal vein thrombosis. AB - OBJECTIVE: To study the prevalence of thrombophilic conditions in patients with acute and chronic portal vein thrombosis (PVT) and to compare it with those in patients suffering from deep vein thrombosis (DVT) after lower limb arthroplasty and in healthy subjects. METHODS: Twenty-six patients with spontaneous PVT (20 chronic, 6 acute) with normal liver function and not receiving anticoagulants were evaluated for thrombophilic conditions. Levels of protein C, protein S and antithrombin were compared with those in 50 healthy controls. Factor V gene 'Leiden' mutation (FVL) and high homocysteine levels were looked for in patients with PVT and in 18 patients developing post-arthroplasty lower limb DVT despite anticoagulation. RESULTS: Of 26 patients with PVT, 19 had at least one thrombotic condition (acute PVT 5/6, chronic PVT 14/20) and 12 had more than one such condition; in comparison, of 18 patients with DVT, eight had one thrombophilic condition and one had two such conditions (p=0.03). Patients with PVT had significantly lower levels of protein C, protein S and antithrombin than healthy subjects and those with DVT. Six patients had Factor VIII levels above 150%; four had elevated homocysteine levels and three had detectable anti-cardiolipin antibodies. Three patients with PVT (acute 2, chronic 1) were heterozygous for FVL mutation. CONCLUSIONS: Underlying thrombophilic conditions are common in Indian patients with spontaneous PVT. In many patients, multiple thrombophilic conditions are present and these may play a role in the pathogenesis of PVT. PMID- 16361766 TI - Low frequency of factor V Leiden and prothrombin G20210A mutations in patients with hepatic venous outflow tract obstruction in northern India: a case-control study. AB - BACKGROUND: Factor V Leiden (FVL) and prothrombin gene (G20210A) mutations are known to be associated with venous thromboembolism. Several studies have shown an association of these mutations with hepatic venous outflow tract obstruction (HVOTO). We studied the prevalence of these mutations among patients with HVOTO in northern India in comparison with healthy population. METHODS: Genomic DNA from patients with HVOTO and healthy controls was analyzed for the presence of FVL and prothrombin gene G20210A mutations, using PCR and restriction-fragment length polymorphism. RESULTS: Fifty-nine patients with HVOTO (age 5-69 years, median 27; 39 male) and 49 unrelated healthy controls from the same geographic region were studied. Of the 59 patients, 19 had a block in the hepatic vein, 7 in inferior vena cava, and 33 had mixed block. Presentation was with acute thrombosis in 9 patients and with long-standing obstruction in 50 patients. Among 49 controls, heterozygous and homozygous FVL mutations were observed in 2 and 0 subjects, respectively, with an allele frequency of 2% (2 of 98). In comparison, among 59 patients with HVOTO, four had heterozygous and none had homozygous FVL mutation, with an allele frequency of 3.4% (p=ns versus controls). The G20210A prothrombin gene mutation was not found in any of the patients or controls. CONCLUSION: FVL and prothrombin G20210A mutations appear to have no role in the pathogenesis of HVOTO in our patients with Budd-Chiari syndrome, consisting largely of those with long-standing obstruction of the inferior vena cava. PMID- 16361767 TI - Benign cystic teratoma of ovary perforating into small intestine with co-existent typhoid fever. AB - Intra-peritoneal rupture or perforation into adjacent abdominal viscera is a rare complication of benign cystic teratoma (dermoid cyst) of the ovary. We report a 48-year-old woman in whom an ovarian dermoid cyst perforated into a loop of the small bowel. The patient had co-existing typhoid fever, which probably predisposed to this complication. She recovered on antibiotic therapy and after surgical excision. PMID- 16361768 TI - Guillain-Barre syndrome associated with acute hepatitis E. AB - A 58-year-old woman presented with a brief icteric illness followed by progressive bilateral symmetrical hypotonic areflexic muscular weakness and unilateral infranuclear facial palsy. She was diagnosed to be suffering from Guillain-Barre syndrome and acute hepatitis E. Such an association has not been described till date. PMID- 16361769 TI - Triple A syndrome. AB - Triple A syndrome (Allgrove syndrome) is an autosomal recessive disorder consisting of achalasia, alacrima and Addison insufficiency. We report an 11-year old girl with predominant symptom of achalasia who was diagnosed as Triple A syndrome almost 3 years after initial presentation. PMID- 16361770 TI - Strangulated femoral hernia presenting as parietal wall emphysema. AB - Surgical emphysema of abdominal and thoracic wall, along with features of intestinal obstruction, has not been reported as a complication of strangulated femoral hernia. We report a 31-year-old woman with such a presentation. PMID- 16361771 TI - Late rejection after transabdominal pre-peritoneal inguinal repair: laparoscopic extraction of mesh. AB - We report a 29-year-old man who developed mesh rejection 3 years after laparoscopic transabdominal pre-peritoneal inguinal repair. The mesh, which was lying in a fluid cavity adherent to the urinary bladder and right iliac vessel, was removed laparoscopically. PMID- 16361772 TI - Primary gastric lymphoma and Helicobacter pylori infection with gastric amyloidosis. AB - Primary gastric lymphoma, an uncommon gastric tumor caused by infection with Helicobacter pylori, is rarely associated with gastric amyloidosis. Chronic bacterial infection is known to cause amyloidosis. We report a 53-year-old man who had an antral and duodenal mass with narrowing and ulceration on endoscopy and CT scan; endoscopic biopsy revealed gastric amyloidosis. Rapid urease test and serology for H. pylori were positive. Histology of resected specimen of distal stomach revealed primary gastric lymphoma, amyloid deposits and spiral organisms suggestive of H. pylori. Rectal biopsy was negative for amyloid. He remained well on follow-up after surgery and eradication of H. pylori. PMID- 16361773 TI - Signet ring cell carcinoma of ampulla of Vater in a young adult. AB - Signet ring cell carcinoma of the ampulla of Vater is extremely rare. The 7 cases reported earlier have been in older patients. We report a 32-year-old lady with this condition, who also had metastases in the bone marrow, vertebrae, lungs and liver. PMID- 16361774 TI - Efficacy and safety of intravenous esomeprazole in gastro-esophageal reflux disease and erosive esophagitis in Indian patients. PMID- 16361775 TI - Esophageal cancer in northeastern Iran. PMID- 16361776 TI - Irritable bowel syndrome: an epidemiological study from the west of Iran. PMID- 16361777 TI - Dysphagia due to transmural migration of polypropylene mesh into esophagus. PMID- 16361778 TI - Isolated tuberculosis of gastric cardia presenting as perforation peritonitis. PMID- 16361779 TI - Clinically significant air entrapment in bowel after percutaneous endoscopic gastrostomy under general anesthesia. PMID- 16361780 TI - Paralytic ileus due to strongyloidosis. PMID- 16361781 TI - [Application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study]. AB - The changes in PSII performance of the photosynthetic apparatus caused by environmental stress or senescence have been explored widely by applying the chlorophyll fluorescence technique, mainly by using the modulated fluorometer. In recent years, an alternative approach via analyzing chlorophyll a fluorescence transient using the continuous excitation fluorometer has been developed, which offers more information that cannot be obtained by the modulated fluorometry. The JIP-test based on the theory of energy fluxes in biomembranes has been widely used to analyze the chlorophyll a fluorescence kinetics transient helping researchers to have a deeper insight into the primary photochemical reaction in the photosynthetic apparatus. In this paper, the chlorophyll a fluorescence transient and its significance, the parameters involved and their significance in the JIP-test, and the application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study are introduced combining some results of the author's. PMID- 16361782 TI - [Progresses in the study on light harvesting pigment protein complexes and reaction centers from purple bacteria]. AB - This review describes the recent progress in understanding of light harvesting complexes and reaction centers from purple bacteria. Emphasis is paid on the structure of two light harvesting complexes, inner or outer, and the mechanism of the transfer of excited energy among relative pigments (Fig.1). At the same time, it is detailedly stated about the understanding of the structure of the reaction center and the transform mechanism from light energy to chemical energy, usable for life system (Fig.2). PMID- 16361783 TI - [Establishment of promoter trapping system mediated by activator/dissociation (beta-glucuronidase) construction in rice]. AB - The coding region of Bar gene, the left border of Ds element, the coding region of GUS gene, the transposase of Ac element, the right border of Ds element and the promoter of Ubi gene were inserted into the T-DNA region of vector pCAMBIA1300 in turn to construct plasmid p13B. The orientations of the ubiquitons' promoter, Ac transposase and Bar are identical but opposite to that of the GUS gene (Fig.1). The plasmid p13B was then introduced into the calli of Oryza sativa subsp. japonica cv. Zhonghua 11 by Agrobacterium tumefaciens mediated transforming to trap genes in rice. Eighteen independent transgenic lines were obtained and propagated. T(2) generations of 18 independent transgenic lines were screening by herbicide (Basta) (Fig.2) and the herbicide-resistant plants obtained were analyzed by PCR (Fig.3). Ds element transposed in an inheritable manner was found in 37 plants, in which 5 plants showed GUS activity (Fig.4). PMID- 16361784 TI - [Construction of tobacco chloroplast multicistron site integration expression vector and its transgene]. AB - According to the published DNA sequence, a serial of elements for constructing the tobacco chloroplast multicistron site integrating expression vectors have been cloned by PCR technique, which include Prrn (a modified plastid ribosomal RNA operon promoter), psbA3' (the 3' region of the plastid psbA gene), aadA gene (encoding aminoglycoside 3'-adenylytransferase), man gene (encoding mannase), gfp gene (encoding green fluorescence protein) and tobacco chloroplast high-frequency homologous recombination ctDNA fragment (psaA/psbC, 3463 bp) (Fig.2). A tobacco chloroplast multicistron expression vector pLM4 (Fig.1) (-psaA-Prrn-SD-man-SD-gfp SD-aadA-psbA3'- psbC-) was constructed with these elements. Then the tobacco leaves were bombarded 5 times with gold particles coated with the vector pLM4. After growing on the screening medium, the function of aadA gene was identified (Fig.3), and the function of gfp gene was confirmed by laser scanner (Fig.4), the expression of man was identified by Western blot (Fig.5). All these genes man, gfp and aadA being integrated in the tobacco chloroplast genome DNA were confirmed by PCR (Fig.6). And the multicistron expression cassette integrating in tobacco chloroplast genome DNA was confirmed by RFLP (Fig.7). All these showed that the three genes in the tomato vector pLM4 were expressed in tobacco chloroplast genome DNA. PMID- 16361785 TI - [Heterologous expression and functional identification of a chitinase gene from wheat]. AB - Chitinases are functional in plant defense against pathogens and also involved in plant development as well as human disease responses. In this study a chitinase gene, Wch2, from wheat was expressed transiently in tobacco leaves and stably in transgenic Arabidopsis thaliana plants to evaluate its heterologous expression and potential application. Western blot analysis revealed that the expressed wheat chitinase was about 30 kD as was expected (Fig.1) and had hydrolytic activity towards chitin polymers (Fig.2). Integration and expression of Wch2 gene in transgenic Arabidopsis were confirmed by PCR (Fig.3) and Western blot (Fig.4) analyses. Wch2 accumulated to a high level in transgenic plants without any detectable degradation. Transgenic plants expressing Wch2 displayed a much higher resistance to Fusarium oxysporum f. sp. matthiolae, a fungus infectious to Arabidopsis thaliana than the plants containing green fluorescence protein (GFP) (Fig.5). These results indicated that Wch2 was suitable for heterologous expression in different organisms and could be used for enhancement of plant resistance in genetic engineering. The Arabidopsis-Fusarium interaction may be one of the ideal plant-fungus interaction systems for the evaluation of antifungal proteins in planta. PMID- 16361786 TI - [Genetic analysis and mapping of the rice leafy-hull mutant Oslh]. AB - Oslh (lh=leafy hull), in the japonica cultivar 9522 background, a mutant of Oryza sativa L. spp. japonica cv. 9522 identified from an M(2) population, was mutagenized by irradiation with (60)Co gamma-ray. The Oslh mutant plants flowered about 15 days later than the wild-type plants (Fig.1e). The paleas, lemmas and lodicules of the flowers of Oslh mutant were transformed into leaf-like structures (Fig.1b, d). Genetic analysis of the F(2) progeny from a cross between the Oslh mutant and wild-type japonica cv. 9522 revealed that the Oslh mutant arouse from a single recessive nuclear gene mutation of the cv. 9522. To map the Oslh locus, an F(2) population generated by crossing between Oslh (japonica) mutant and Guangluai4 (indica) was analyzed. The Oslh locus was mapped to the long arm of rice chromosome 3, between a SSR marker RM5475 and an InDel marker GY305, 2.9 and 1.5 cM away from these two markers respectively (Fig.4). These results are useful for further cloning and functional analysis of the OsLH gene. PMID- 16361787 TI - Activation of genes presumed in cytokinin signal pathway of two-component system in response to the increased cytokinin contents in ipt-GUS transgenic Arabidopsis. AB - The ipt-GUS activated transgenic Arabidopsis had 20-25 fold higher cytokinin contents than the wild type. Changes in cytokinin content in vivo triggering gene expressions involved in signal pathway of two-component system have been analyzed on the day of 6 d, 12 d, 20 d and 30 d after seed cultivation on MS medium in light conditions. The results showed that the two cytokinin receptors, His-kinase CRE1 was more sensitive to the increased cytokinin contents than CKI1. Arabidopsis response regulators, ARR4 and ARR5, were induced by the increased cytokinin contents at different time after seed germination. ARR4 responded to cytokinins at early time of seed germination, especially on the 6 d when seedlings were around true leaf initiation, while cytokinins induced ARR5 activation after 6 d of seed cultivation in light conditions, an obvious increase was on the 20 d when seedlings were around inflorescent shoot initiation. Hpt type transmitter kinase AHP4 increased its activation by cytokinin induction only between the 20 d and 30 d after seed cultivation in light conditions, and an obvious increase was on the 20 d. PMID- 16361788 TI - Stress-induced Rim2/Hipa pseudogene of rice exhibits alternative tailing and splicing during transcription. AB - The rice Rim2/Hipa is a unique stress-induced transposon superfamily recently identified in Oryza genomes. In the present study, we conducted genome-wide screening of full-length Rim2 cDNA from the pathogen-induced cDNA libraries and mining of cDNA databases. Four indica and two japonica types of transcripts were identified, which were transcribed from the same Rim2 pseudogene Rim2-42 that contains premature stop codons in the TNP2-TPase coding region. These data demonstrated that the processing of the Rim2 transcripts exhibited variations within and between the two subspecies. These transcripts were found to be produced by alternative transcription (tailing) or splicing from Rim2-42 under stress conditions. An additional Rim2-like transcript (Rim2-XET), a chimera of Rim2 and XET genes, were also found to be derived from read-through. These results show that the Rim2 transposon probably loses its transposition capacity during evolution, and that Rim2-42 inserts downstream of the stress-inducible XET promoter, resulting in Rim2 transcript accumulation upon pathogen attack. PMID- 16361789 TI - [Calcium distribution in fertile and sterile anthers of a genic male sterile Chinese cabbage]. AB - Potassium antimonite was used to locate calcium in the fertile and sterile anthers of a genic male sterile Chinese cabbage (Brassica campestris L. ssp. chinensis Makino) to probe the relation between Ca(2+) and fertility and sterility of anthers of the cabbage. During fertile anther development, calcium granules increase in number in anther wall cells after meiosis, and then appeared also in locule, suggesting a calcium influx into locule from anther wall cells (Plate I-4). Then the number of calcium granules in microspore cytoplasm also increased at early stage (Plate II-1), accumulated mainly on the membrane of small vacuoles which were fusing to form big ones to make a polarity in the cell and to prepare asymmetric division of microspore (Plate II-3,4). After microspore division and the big vacuole decomposition, many calcium granules accumulated again on the membrane of the vacuoles (Plate III-1,2), displaying calcium regulates vacuole formation and decomposition during pollen development. In sterile anthers, abnormal distribution of calcium granules first appeared in callus wall of microspore mother cell (Plate IV-1). However, only a few calcium granules appeared in early microspores, which then could not form small vacuoles and finally a big vacuole (Plate IV-2,3). The aborting microspores degenerate by cytoplasm shrinking (Plate IV-5,6). The difference pattern of distribution of calcium granules between the fertile and sterile anthers indicates that anomalies in the distribution of calcium accumulation are correlated with the failure of pollen development and pollen abortion. PMID- 16361790 TI - [Reducing the maize amylopectin content through RNA interference manipulation]. AB - To regulate the biosynthesis of maize starch, gene segment of starch branch enzyme (SBE) in maize was cloned and an expression vector of small interference RNA with the fragment of antisense gene + sense gene (pCJSBE2b, Fig.1) was constructed. The expression vector of pCJSBE2b was transformed into maize inbred lines by using pollen-tube pathway. Results of PCR amplification and Southern blotting indicated that the aimed gene with the fragment of antisense gene + sense gene had integrated into receptor maize genome (Fig.4,5) and inherited steadily in the transgenic plants (Table 1). Northern hybridization analysis showed that the aimed gene transcribed normally in the transgenic plants, which resulted in a reduction of endogenous SBE mRNA (Fig.6). The analysis of the activity of starch branching enzyme showed that the enzyme activity decreased obviously in the transgenic plants and maximum decrease was 85% compared to the control plants (Fig.7a). Although the total starch content of transgenic plants showed no change compared to the control plants but the amylose content increased to 50% (Fig.7b) in transgenic plants about 27% higher than receptor maize plants. PMID- 16361791 TI - [Genetic analysis of starch branching enzyme activity in rice grain]. AB - To make an genetic analysis of starch branching enzyme (Q enzyme) inheritance, which is important in catalyzing the formation of amylopectin and thereby raising the amylopectin/amylose ratio of starch in rice grains, a linkage map of 207 DNA markers were constructed by using 247 recombinant inbred lines derived from an indica-indica rice cross Zhenshan 97B/Milyang46. In 2002 and 2003, the activities of starch branching enzyme of the parents and 247 RILs were measured 10 d and 20 d respectively after flowering (Table 1). A total of 3 quantitative trait loci (QTLs) were detected to have significant additive effects on Q enzyme activities 10 d after flowering with 10% phenotypic variations explained for the three QTLs (Table 2, Fig.1). Meanwhile, qQ10-6 with significant QTL x environment interaction was detected. Epistasis analysis detected 5 and 2 significant additive-by-additive interactions for Q enzyme 10 d and 20 d respectively after flowering (Table 3), and three pairs of QTLs 10 d after flowering with significant epistasis x environment interactions were detected, explaining 3% to 12% of the phenotypic variation. The results showed that the environmental factors had obvious effect on the gene expression of Q enzyme activity in rice grain. Meanwhile, the location of qQ10-6 in relation to that of QTLs for gelatinization temperature, gene loci involved in ADPG pyrophosphorylase and soluble starch synthase were discussed with the help of a genetic map. PMID- 16361792 TI - [Effects of exogenous polyamines on the growth and activities of H+-ATPase and H+ PPase in cucumber seedling roots under hypoxia stress]. AB - The effects of exogenous polyamines on the growth, polyamines contents, the activities of H(+)-ATPase and H(+)-PPase of plasma membrane and tonoplast in cucumber seedling roots under root-zone hypoxia stress were studied under hydroponic culture. The results showed that the growth of cucumber roots was significantly inhibited by hypoxia stress (Table 1). While the growth inhibition in cucumber roots by hypoxia stress could be alleviated by spraying exogenous polyamines. From these results (Table 1 and 2) we can conclude that Spd played an active physiological role in relieving hypoxia stress and Put exerted actively physiological role by enhancing Spd biosynthesis. The results also showed that the endogenous polyamine contents in cucumber roots were increased by hypoxia stress and spraying exogenous polyamines. Exogenous polyamines application obviously promoted the H(+)-ATPase activities of plasma membrane in cucumber roots (Fig.1), but did not significantly affect the H(+)-ATPase and H(+)-PPase activities of tonoplast in cucumber roots (Fig.2,3). The results suggested that exogenous polyamines could enhance hypoxia resistance of cucumber seedlings by increasing H(+)-ATPase activities of plasma membrane in cucumber roots. PMID- 16361793 TI - [Changes in some defensive enzyme activity induced by the piercing-sucking of brown planthopper in rice]. AB - The effects of brown planthopper on the activities of some key enzymes involved in defense response and the H(2)O(2) content were investigated in rice. From the fact that after piercing-sucking of brown planthopper, malondialdehyde (MDA) content of rice plant was increased rapidly (Fig.3), one can conclude that it induced lipid peroxidation; lipoxygenase (LOX), hydroperoxide lyase (HPL) and phenylalanine ammonia-lyase (PAL) activities were increased significantly in plants of resistant and susceptible varieties by piercing-sucking of brown planthopper (Figs.1,2,4); whereas the catalase activity and the H(2)O(2) content were significantly different between resistant and susceptible plants by piercing sucking of brown planthopper, the former being decreased and the latter increased in resistance plants of variety RH (Oryza sativa L. cv. Rathu Heenati), while the reverse was true in the susceptible plants of variety TN1 (Oryza sotiva L. cv. Taichung Native 1) (Figs.5,6). The effect of brown planthopper on the PAL activity and the H(2)O(2) content were systemic (Figs.4,6), but the effect on the LOX activity occurred only in stem, the organ which brown planthopper sucking (Fig.1). PMID- 16361794 TI - [Changes in cell wall component metabolism and ultrastructure of postharvest persimmon fruit during softening]. AB - The changes in cellular wall hydrolases, cellular wall components and cellular wall ultrastructure of postharvest persimmon (Diospyros kaki L. cv. Bianhua) fruit during softening were studied. Pectinesterase activity increased sharply at first and reached a peak (Fig.3A). Significant correlation was observed between the pectinesterase activity and the loss of flesh firmness (r= -0.74). Polygalacturonase activity increased slowly (Fig.3B), but there was no significant correlation between the polygalacturonase activity and the loss of flesh firmness. Beta-galactosidase activity increased sharply (Fig.3C), with a negative correlation between the b-galactosidase activity and the loss of flesh fruit firmness (r= -0.77). Cellulase activity increased markedly during ripening (Fig.3D). Significant correlation was observed between cellulase activity and the loss of flesh firmness (r= -0.90). Consistent with the increases in activity of cell wall hydrolases of cell wall constituents, fruit softening was accompanied by a progressive increase in WSP (water soluble pectin) content and a progressive decrease in protopectin and cellulose content (Fig.4). The cell wall structure was integrated when persimmon was harvested (Fig.5A). After 3 d of ripening, the middle lamella became liquefied (Fig.5B), or even the primary cell wall was dissolved in some regions (Fig.5C). PMID- 16361795 TI - [Effects of high temperature on grain filling and some physiological characteristic in flag leaves of hybrid rice]. AB - The effects of high temperature on grain filling rate and some physiological indexes in flag leaves in hybrid rice Teyou 559 were studied. The results showed that chlorophyll content and superoxide dismutase (SOD) activity in flag leaves were lower significantly by high temperature stress (Figs.2A and 3A). The electrolyte leakage and malondialdehyde (MDA) content were higher significantly under high temperature stress (Figs.2B and 3B). The proline (Pro), ascorbic acid (AsA), glutathione (GSH), soluble protein and soluble sugar contents were lower significantly by high temperature stress than those of the control (Figs.4,5 and 6). Both the rate of grain filling and grain dry matter accumulation become lower significantly under high temperature stress (Figs.7 and 8). Therefore, it was suggested that high temperature accelerated the senescence of rice leaf and decreased the photosynthesis rate during grain filling period, and then which was the main reason in physiology which resulted the decrease of grain filling rate, seed setting rate, grain weight and yield. PMID- 16361796 TI - [Changes in amino acid and fatty acid contents as well as activity of some related enzymes in apple fruit during aroma production]. AB - Aroma volatiles from apple (Malus domestica Borkh. var. Starkrimson) fruit at different stages of maturity were collected by solid adsorbent-Tenax-GC and determined by thermodesorption and GC-MS. Production of propyl acetate, butyl acetate, ethyl 2-methyl-butanoate and total ester volatiles and changes in concentration of the precursors of aroma biosynthsis--free amino acids and fatty acids and activities of lipoxygenases (LOX) and alcohol acetyltransferase (AAT) in apple fruits during ripening were studied. The results showed that propyl acetate and total esters were very low when the endogenous ethylene formation of the fruit was very low. At the stage of the increase in ethylene production, the rate of formation of propyl acetate and total esters increased. Butyl acetate appeared at the beginning of ethylene rise and increased thereafter. Ethyl 2 methyl-butanoate was produced at the beginning of climacteric stage and then increased sharply (Figs.1). These facts suggest that the aroma production is closely related to ethylene production. Among the 14 free amino acids detected in fruit, isoleucine which is considered to be the biosynthetic precursor of some branched chain esters showed a great increase during fruit ripening while the others decreased or remained stable (Table 1). The accumulation of isoleucine suggested that isoleucine supply in fruit may not limit the biosynthesis of esters with branched chain alkyl groups. Concentrations of free fatty acids such as palmitic, linolenic, oleic, linoleic, stearic acids increased before the increase of aroma production, decreased with the increase of aroma production and showed an increase at postclimacteric stages (Fig.2). LOX activity increased at climacteric stages and declined rapidly thereafter. AAT activity increased sharply at the early stage of fruit maturity when the aroma was very low and remained at a stable high level during fruit ripening (Fig.3) indicating that the AAT activity is not the limiting factor for aroma formation in apple fruit. PMID- 16361797 TI - Diagnosis and treatment of nontuberculous mycobacterial pulmonary diseases: a Korean perspective. AB - The incidence of pulmonary disease caused by nontuberculous mycobacteria (NTM) appears to be increasing worldwide. In Korea, M. avium complex and M. abscessus account for most of the pathogens encountered, whilst M. kansasii is a relatively uncommon cause of NTM pulmonary diseases. NTM pulmonary disease is highly complex in terms of its clinical presentation and management. Because its clinical features are indistinguishable from those of pulmonary tuberculosis and NTMs are ubiquitous in the environment, the isolation and identification of causative organisms are mandatory for diagnosis, and some specific diagnostic criteria have been proposed. The treatment of NTM pulmonary disease depends on the infecting species, but decisions concerning the institution of treatment are never easy. Treatment requires the use of multiple drugs for 18 to 24 months. Thus, treatment is expensive, often has significant side effects, and is frequently not curative. Therefore, clinicians should be confident that there is sufficient pathology to warrant prolonged, multidrug treatment regimens. In all of the situations, outcomes can be best optimized only when clinicians, radiologists, and laboratories work cooperatively. PMID- 16361798 TI - Leukotriene-related gene polymorphisms in patients with aspirin-intolerant urticaria and aspirin-intolerant asthma: differing contributions of ALOX5 polymorphism in Korean population. AB - The pathogenesis of aspirin (acetylsalicylic acid, ASA)-intolerant urticaria (AIU) is still poorly understood but it has recently been suggested that it is associated with the overproduction of leukotriene (LT). This is supported by evidence that cyclooxygenase 2 inhibitor is given safely to patients with AIU. The present study was designed to investigate the role of genetic polymorphism of LT related genes in the pathogenesis of AIU via a case-control study. We screened single nucleotide polymorphisms (SNPs) in genes encoding enzymes involved in leukotriene synthesis in the Korean population with AIU (n = 101), ASA-intolerant asthma (AIA, n = 95) and normal healthy controls (n = 123). Genotype was determined by primer extension reactions using the SNapShot ddNTP primer extension kit. Among 8 SNPs of four LT related genes, the polymorphism of ALOX5 at positions of -1708 G > A showed significant difference in genotype frequency between AIU and AIA (p = 0.01). Furthermore, there were significant differences observed in the frequencies of two ALOX5 haplotypes between the AIU group and AIA group (p < 0.05). However, there were no differences in allele, genotype, or haplotype frequencies of ALOX5 between the AIU group and the normal control group. These results suggested that ALOX5 has a differing contribution in two major clinical pathogenesis related to ASA-sensitivity. PMID- 16361799 TI - Increased anti-tumor effect by a combination of HSV thymidine kinase suicide gene therapy and interferon-gamma/GM-CSF cytokine gene therapy in CT26 tumor model. AB - The potential therapeutic benefit of introducing IFN-gamma and GM-CSF genes in combination with the HSVtk suicide gene into subcutaneously implanted CT26 tumor cells was compared with that from each treatment alone. Cells, unmodified or retrovirally transduced with HSVtk or IFN-gamma/GM-CSF genes, were inoculated subcutaneously into syngeneic BALB/c mice in various combinations. HSVtk gene, with intraperitoneal ganciclovir treatment, reduced tumor volume by 81% at locally inoculated tumor sites (p < 0.01) and by 25% at distantly inoculated tumor sites (p = 0.052). IFN-gamma/GM-CSF genes showed a 56% tumor volume reduction at local tumor sites (p < 0.01) and 15% volume reduction at remote tumor sites, although this was not statistically significant. The combination of HSVtk (with GCV) and IFN-gamma/GM-CSF genes showed an 81% volume reduction at local tumor sites (p < 0.01) and a 43% volume reduction at remote tumor sites (p < 0.01). Thus, the combination of HSVtk and IFN-gamma/GM-CSF gene therapy produced greater therapeutic efficacy than either treatment alone. PMID- 16361800 TI - Asymptomatic infection by Streptococcus pyogenes in schoolchildren and diagnostic usefulness of antideoxyribonuclease B. AB - This study is designed to evaluate the immune status of schoolchildren with respect to Streptococcus pyogenes, and to ascertain the usefulness of antideoxyribonuclease B (ADNase B). Antistreptolysin O (ASO) and ADNase B concentrations were measured quantitatively in 266 serum samples from healthy elementary school children in Seoul. Simultaneously, throat cultures were taken in order to isolate S. pyogenes and other beta-hemolytic streptococci (BHS). The upper limits of the normal (ULN) concentration of ASO and ADNase B were 326 IU/mL, and 362 IU/mL, respectively. The correlation between ADNase B (y) and ASO (x) was y = 0.4x+173 (r = 0.46). Mean ADNase B level (392 IU/mL) was significantly higher in children with S. pyogenes than in those with non-group A BHS (236 IU/mL) or no BHS (234 IU/ mL). Some schoolchildren were proven, via ASO and ADNase B tests, to be harboring asymptomatic S. pyogenes infections. The high ULN of ASO and ADNase B in schoolchildren should be carefully considered, in order to interpret the data collected from the patients. We could add the ADNase B test to our set of diagnostic tools, which would allow us to more accurately detect and diagnose streptococcal infections, as ADNase B was more specifically related to the results of throat cultures, and there was little correlation between ASO and ADNase B. PMID- 16361801 TI - A sporadic outbreak of human brucellosis in Korea. AB - Eleven cases of human brucellosis occurred among livestock workers and a veterinarian who lived and worked in a rural area around Jeongeup City, Jeollabuk Do, Korea from February 2003 to August 2003. Eight of the patients had taken care of Korean native cattle that were infected with bovine brucellosis and had already been slaughtered. Two of the patients had taken care of dairy cattle, and one case was a veterinarian who acquired the disease through an accidental contact with infected cattle while assisting in calf delivery. Eleven cases were identified by serologic work ups and four cases were identified via positive blood cultures. This study shows that the Republic of Korea is no longer free of human brucellosis, Brucella abortus biotype 1. We reviewed the patients' characteristics and serologic data during the one-year follow up period, and we also discuss on the efficacy and side effects of the rifampin and doxycyline regimen used for the treatment of human brucellosis. PMID- 16361802 TI - Somatostatin for postoperative chylothorax after surgery for children with congenital heart disease. AB - Chylothorax is a rare but serious postoperative condition with a high rate of morbidity that may lead to death of children with congenital heart disease. Here we reviewed nine consecutive cases with chylothorax in infants and children following cardiac surgery from March 2002 to February 2003. Somatostatin was added to conservative treatment protocol to increase effectiveness of therapy in all cases. The duration of somatostatin therapy varied from 7 to 32 days. All cases of chylothorax were successfully treated with intravenous infusion of somatostatin as an adjunctive treatment. Even though two cases showed rebound phenomena, we avoided any surgical procedure in the nine patients who treated with conservative management combined with somatostatin. No significant side effects of somatostatin were observed. It seems that somatostatin is effective, noninvasive and safe therapeutic modality. It can be used as an adjunctive treatment to conservative management to control postoperative chylothorax in children with congenital heart disease. PMID- 16361803 TI - Immunoreactivity of CD99 in non-Hodgkin's lymphoma: unexpected frequent expression in ALK-positive anaplastic large cell lymphoma. AB - To verify the spectrum of CD99-expressing lymphoid malignancy, an immunohistochemical study for CD99 was carried out in 182 cases of non-Hodgkin's lymphoma, including 21 lymphoblastic lymphomas, 11 small lymphocytic lymphomas, 9 mantle cell lymphomas, 12 follicular lymphomas, 37 diffuse large B cell lymphomas, 18 Burkitt's lymphomas, 28 NK/T-cell lymphomas, 8 angioimmunoblastic T cell lymphomas, 23 peripheral T-cell lymphomas, unspecified, and 15 systemic anaplastic large cell lymphomas. CD99 was positive in all T-lymphoblastic lymphomas and in 60% of B-lymphoblastic lymphomas. Majority of T and NK cell lymphomas were negative for CD99, except anaplastic large cell lymphomas (ALCLs). Eight of 15 cases (54%) of ALCLs reacted with anti CD99 antibody. Seven of 10 (70%) ALK positive ALCLs expressed CD99, whereas only 1 of 5 (20%) ALK negative ALCLs were positive. Of the mature B-cell lymphomas, 5.4% (2/37) of diffuse large B cell lymphomas and 11.1% (2/18) of Burkitt's lymphomas expressed CD99. In conclusion, CD99 is infrequently expressed in mature B and T cell lymphomas, except ALK-positive ALCL. High expression of CD99 in ALK-positive ALCL is unexpected finding and its biologic and clinical significances have yet to be clarified. PMID- 16361804 TI - Mycobacterium kansasii pulmonary diseases in Korea. AB - Mycobacterium kansasii is one of the most common cause of pulmonary diseases due to nontuberculous mycobacteria. We investigated the changing in the number of isolation of M. kansasii and the clinical characteristics of M. kansasii pulmonary disease in Korea. Through searching the database of the Korean Institute of Tuberculosis, we identified the cases of isolated M. kansasii from 1992 to 2002. The number of M. kansasii isolation had increased from once in 1992 to 62 in 2002. Fifteen patients with M. kansasii pulmonary disease were identified during the period January 1997 to December 2002. Twelve patients (80%) were male and fourteen (93%) were from highly industrialized areas. The most common symptom was a cough. Seven patients (47%) had a cavitary lesion and right upper lobe was most commonly involved. Patients responded well to isoniazid and rifampicin based regimens both bacteriologically and radiographically. In conclusion, M. kansasii isolation has increased, especially in highly industrialized areas, as well as other nontuberculous mycobacteria in Korea. PMID- 16361805 TI - Emergence and wide dissemination of CTX-M-type ESBLs, and CMY-2- and DHA-1-type AmpC beta-lactamases in Korean respiratory isolates of Klebsiella pneumoniae. AB - Respiratory isolates of Klebsiella pneumoniae in Korea during 2002-2003 were studied to determine the prevalence and types of extended-spectrum beta lactamases (ESBLs) and plasmid-mediated AmpC beta-lactamases (PABLs). ESBL production was tested by double-disk synergy, and genotypes of beta-lactamases were determined by PCR and sequencing. ESBLs were detected in 28.4% of 373 isolates, and the most prevalent types were SHV-12 (63 isolates) and CTX-M-14 (9 isolates). Forty of 75 ESBL-producers (53.5%) also had PABLs: 21 isolates with CMY-2-like, 17 with DHA-1-like. Pulsed-field gel electrophoresis showed 19 types and 25 of 74 isolates had an identical pattern, indicating nosocomial spread. Dissemination of ESBL- and PABL-producing K. pneumoniae strains in Korea is a particular concern, as it limits the choice of antimicrobial agents for treatment of infections. PMID- 16361806 TI - Biweekly irinotecan and cisplatin as second-line chemotherapy in pretreated patients with advanced gastric cancer: a multicenter phase II study. AB - The current phase II study was conducted to evaluate the response rate and safety of a combination regimen of biweekly irinotecan plus cisplatin in pretreated patients with advanced gastric cancer. Patients with previously treated metastatic or recurrent gastric cancer received intravenous irinotecan 70 mg/m2 and cisplatin 30 mg/m2 on day 1 and 15 every 4-week cycle. Thirty-two patients were enrolled in the current study. Of these, 31 patients were assessable for efficacy and all for toxicity. No complete response and 5 partial responses were confirmed, giving an overall response rate of 15.6% (95% CI; 2.3-28.9%). The median time to progression and median overall survival for all patients was 113 days and 184 days, respectively. Grade 3/4 neutropenia occurred in 6 patients (18.8%), yet no febrile neutropenia was observed. In addition, grade 3 anorexia was observed in 4 patients (12.5%) and grade 3 diarrhea occurred in 2 patients (6.2%). The combination chemotherapy of biweekly irinotecan and cisplatin was found to be moderately effective and well tolerated in pretreated patients with advanced gastric cancer. Accordingly, this regimen can be regarded as an important second-line treatment option for advanced gastric cancer. PMID- 16361807 TI - Comparison of Behcet's disease and recurrent aphthous ulcer according to characteristics of gastrointestinal symptoms. AB - Behcet's disease (BD) is a multisystemic chronic inflammatory disease. It is characterized by recurrent oral and genital ulcers, uveitis, skin lesions and other manifestations, including neurologic, vascular, joint, and gastrointestinal ulcers of variable severity. Recurrent aphthous ulcer (RAU) represents a very common, but poorly understood, mucosal disorder. If a patient of RAU without any other typical symptoms of BD has gastrointestinal symptoms, it is difficult to distinguish this RAU from true BD with gastrointestinal involvement. Because pathognomonic clinical features and tools are absent, the differential diagnosis of these two diseases relies on the characteristic clinical features and the judgement of an experienced physician. Sixty-five out of a total 960 RAU patients and forty-four of 556 BD patients with gastrointestinal symptoms between January 1996 and December 2003 participated in this study. All were evaluated with esophagogastroduodenoscopy and colonoscopy. Clinical, endoscopic and histopathologic findings were analyzed and ELISA tests were conducted to detect serum levels of ASCA and pANCA. No significant difference was found between the two groups. Differential diagnosis between RAU with gastrointestinal symptoms and BD with gastrointestinal involvement requires further prospective, large-scale study. PMID- 16361808 TI - Gastrointestinal stromal tumors in Koreans: it's incidence and the clinical, pathologic and immunohistochemical findings. AB - Seven hundred forty seven cases of gastrointestinal stromal tumors (GISTs) in Koreans who were diagnosed between 2001 and 2002 were analyzed to evaluate their occurrence and their clinical, pathologic and immunohistochemical findings. The most frequent location of tumor was in the stomach (63%), followed by the small intestine (30%), the colorectum (5%), and the esophagus (2%). c-kit expression was found in 93.6% of the cases, while CD34, SMA and S-100 protein was positive in 80.1%, 28.2%, and 20.2%, respectively. c-kit positivity was high in the stomach (94.2%) and small intestine (94.6%), while it was relatively low in the colorectum (85.0%), and esophagus (81.2%). The positivity for CD34 was correlated with the higher risk of GISTs (p = 0.04). Follow up of the patients showed that 58 primary GISTs patients died and 20 of these patients were recurrent or metastatic at the time of diagnosis. The pathologic diagnosis to predict the risk of aggressive behavior of GISTs was correlated with the numbers of tumor, clinical stage, epithelioid histologic type, cellularity, cellular atypia, necrosis, and mucosal invasion (p = 0.00). GISTs with a poor prognosis were closely related to the clinical stage at presentation, the locations of the tumor, and the ages of the patients. PMID- 16361809 TI - The prevalence of hepatitis C virus infection in Korea: pooled analysis. AB - This study evaluated the prevalence of hepatitis C virus (HCV) infections in Korea. Pooled estimates of the anti-HCV positivity were calculated using the data published in 15 reports on the general population and health check-up examinees. The overall pooled estimate of the prevalence of HCV among middle-aged adults (40 yr old and above) was 1.68% (95% confidence interval: 1.51-1.86%) during the year of 1990-2000 among the general population. Most of the published data indicated that the prevalence of anti-HCV increased with age. The anti-HCV positivity was significantly higher in females than in males. Because the risk of HCV exposure in blood recipients has decreased remarkably, the spread of HCV through means other than a transfusion must be prevented. PMID- 16361810 TI - Interleukin-10 promoter polymorphism is associated with the predisposition to the development of IgA nephropathy and focal segmental glomerulosclerosis in Korea. AB - The roles of interleukin-10 (IL-10) have been emphasized in several models of glomerulonephritis (GN). Three biallelic polymorphisms within the IL-10 promoter region, at positions -1,082, -819, and -592 from the transcription initiation site, were shown to affect the level of IL-10 production. To investigate the effect of IL-10 promoter polymorphisms on the predisposition to development of GN in Korea, IL-10 promoter polymorphisms were assayed by polymerase chain reaction followed by restriction fragment length polymorphism in 108 patients with IgA nephropathy (IgAN), 49 focal segmental glomerulosclerosis (FSGS), and 100 healthy controls. In comparison with the control, the frequency of -1,082*G alleles were lower in IgAN and the frequencies of -592*C and -819*C were lower in FSGS, respectively. As for the haplotype, GCC haplotype was less frequent among IgAN than the control and ATA haplotype was more frequent among FSGS than the control (p < 0.05). The frequency of intermediate producer genotypes (GCC/ACC and GCC/ATA) were lower among IgAN or FSGS than the control. Our findings suggested that IL-10 promoter polymorphism predisposed to the development of IgAN and FSGS in Korean patients. PMID- 16361811 TI - Coronary-artery calcium scores using electron beam CT in patients with chronic renal failure. AB - We evaluated the risk of coronary-artery disease in patients with chronic renal failure (CRF) by measuring the coronary-artery calcium scores with electron beam CT (EBCT). A total of 81 CRF patients were divided into three groups; pre dialysis (group I, n = 35), hemodialysis (group II, n = 31) and peritoneal dialysis (group III, n = 15). The several serum biochemical markers and calcium score levels by EBCT were determined. The Ca x P products were significantly higher in groups II (p < 0.05) and III (p < 0.01) than in group I. The serum calcium levels were significantly higher in group III than in both group I (p < 0.01) and II (p < 0.05). The serum calcium level in 15 patients with a calcium score > 400 was significantly higher than the 66 patients with a score < or =400 (p < 0.01). The calcium score was significantly higher in the 15 patients with cardiovascular complications than in the 66 patients without cardiovascular complications (628.9+/-904.8 vs. 150.4+/-350.9, p < 0.01). EBCT seemed to be a good diagnostic tool for evaluating the risk of coronary-artery disease ''noninvasively'' in CRF patients who are at increased risk of cardiovascular morbidity and mortality. PMID- 16361812 TI - Investigation of early protein changes in the urinary bladder following partial bladder outlet obstruction by proteomic approach. AB - We investigated the pathophysiological mechanism by proteomic approach as a possible tool to detect the marker proteins to develop lower urinary tract symptoms following bladder outlet obstruction (BOO). Rats were randomized into 3 groups; control, sham operation and BOO groups. BOO group was divided into 1, 3, and 5 day-group. Conventional proteomics was performed with high resolution 2-D gel electrophoresis followed by computational image analysis and protein identification using mass spectrometry using rat urinary bladders. A comparison of bladder of BOO group with control bladder showed that three proteins of optineurin, thioredoxin and preprohaptoglobin were over-expressed in the bladder of BOO group. In addition, four proteins, such as peroxiredoxin 2, transgelin, hippocampal cholinergic neurostimulating peptide (HCNP) and beta-galactoside binding lectin, were under-expressed in the bladder of BOO group. These data supported that downregulation of HCNP might make detrusor muscle be supersensitive to acetylcholine, up-regulation of optineurin means the protection of nerve injury, and down-regulation of transgelin means the decreased contractility of detrusor muscle. Beside these proteins, other proteins are related to oxidative stress or have a nonspecific function in this study. However more information is needed in human bladder tissue for clinical usage. PMID- 16361813 TI - Risk factors of voiding dysfunction and patient satisfaction after tension-free vaginal tape procedure. AB - This study was undertaken to identify risk factors for postoperative voiding dysfunction and factors having impact on patient global satisfaction after a tension-free vaginal tape (TVT) procedure. Two hundred and eighty-five women who underwent the TVT procedure for stress urinary incontinence were analyzed to identify risk factors predictive of voiding dysfunction. Postoperative voiding dysfunction was defined as a peak urinary flow rate (PFR) <10 mL/sec (straining voiding, n = 17) or residual urine volume >30% of bladder capacity (incomplete emptying, n = 13). The global satisfaction rate was 91.6%. Voiding dysfunction developed in 29 (10.2%) patients. Among the factors, PFR was only factor of significance for voiding dysfunction. There was no significant difference between patients with and without voiding dysfunction in terms of their satisfaction. But postoperative PFR <10 mL/sec significantly compromised global satisfaction after the surgery. In those patients with a preoperative PFR <20 mL/sec, there were more patients with postoperative PFR <10 mL/sec. Peak urinary flow rate is an important factor for the postoperative voiding dysfunction. The inevitable decline in PFR can compromise patients' satisfaction with the procedure, when their postoperative PFR was <10 mL/sec. PMID- 16361814 TI - Quantitative ultrasound of the calcaneus in a Korean population: reference data and relationship to bone mineral density determined by peripheral dual X-ray absorptiometry. AB - The aim of this study was to establish reference data for the quantitative ultrasound (QUS) of the calcaneus and for the bone mineral densities (BMD) of the calcaneus and distal forearm, and to evaluate the correlation between QUS parameters and BMD in a Korean population. We performed a cross-sectional study involving 3,053 subjects (1,225 men and 1,828 women). QUS was conducted on the calcaneus and was quantified as speed of sound (SOS, m/sec), broadband ultrasound attenuation (BUA, dB/MHz), and stiffness index. The BMD of the calcaneus and distal forearm were measured using dual X-ray absorptiometry. The peak mean values for the QUS parameters occurred in the 20 to 29-yr-old subjects of both sexes, with the exception of the BUA, which reached the highest values in women of 30-39 yr. For both sexes, the mean BMD of the calcaneus was highest in those 20-29 yr old and that of the distal forearm was highest in those 40-49 yr old. The correlations between the QUS and BMD results were found to be 0.41 to 0.73 in men and 0.51 to 0.76 in women. Theses data can serve as a reference values for both sexes in Korea. PMID- 16361815 TI - Cytokine production by whole blood cells: relationship to interleukin gene polymorphism and bone mass. AB - The aims of this study were to investigate the relationships between the production of interleukin-1 (IL-1), and IL-6 system by whole blood cells, and bone mineral density (BMD), and polymorphisms in IL-1 system and IL-6 gene in postmenopausal Korean women. The production of IL-1alpha, IL-1beta, IL-1 receptor antagonist (IL-1ra), IL-6, and soluble IL-6 receptor (sIL-6r) by lipopolysaccharide-stimulated whole blood cells was measured by ELISA in 110 subjects. Serum osteocalcin, C-telopeptide of type I collagen, and BMD at lumbar spine and proximal femur were measured. IL-1alphaC(-889)T polymorphism, IL-1beta C(-511)T polymorphism, 86-base pair variable number tandem repeat polymorphism in the IL-1ra gene, and IL-6 C(-634)G polymorphism were analyzed. The production of IL-1beta correlated positively with BMD at femoral neck, whereas the production of other ILs did not correlate with BMD at the skeletal sites examined. No significant differences in the production of ILs were observed among normal, osteopenic and osteoporotic postmenopausal women, and among the different IL system polymorphisms groups studied. No correlation between bone turnover markers and the production of ILs was noted. In conclusion IL-1beta may regulate bone metabolism at femoral neck, and the IL system polymorphism do not affect the production of ILs by whole blood cells. PMID- 16361816 TI - Percutaneous vertebroplasty and facet joint block. AB - It is surprising that about 24% of patients with benign osteoporotic vertebral fracture die within a year from respiratory infection and urinary tract infection because of coughing and voiding difficulties, depending on the sites of compression fractures. We reviewed 500 patients on whom percutaneous vertebroplasty (PVP) was performed, at 612 levels in terms of patient selection, operation technique, medication, and clinical outcomes during the follow-up course for 2 yr study period. To confirm the most painful level among the multiple fracture sites, physical examination after facet joint block under the fluoroscope was the most reliable method. The mean total lumbar spine fracture threshold of bone mineral density was 0.81+/-0.05 g/cm2. The mean changes of numeric rating scale scores, Oswestry Disability Index except sex life, and Karnofsky performance status were -72.00, -83.50 and +60.62% in the osteoporosis group and -51.89, -45.02, and 69.03% in the tumor group. Complications related to the procedure were lateral spinal cord damage, transient paresthesia and transient hypotension. PVP with facet joint block is a profitable method for the vertebral compression fracture because of low risk and short duration of procedure with a high chance to result in pain relief and early mobilization. PMID- 16361817 TI - The prevalence of metabolic syndrome in patients with gout: a multicenter study. AB - It has been suggested that hyperuricemia and possibly gout are associated with the metabolic syndrome, but there have been no direct studies. This study was undertaken to obtain the prevalence of the metabolic syndrome in patients with gout and to compare it with those from the general population studies. This was a 4-institutional case-historical control study composed of 168 patients with gout. We assessed the prevalence of metabolic syndrome according to the ATP III criteria and compared the prevalence with that of the historical controls. To elucidate the factors in gout that were associated with metabolic syndrome, a multivariate analysis was done. The age-adjusted prevalence of metabolic syndrome in gout patients was 43.6%, which was significantly higher than that of the Korean control population (5.2%) from the previous studies. Patients with gout had more components of metabolic syndrome than did the controls. Body mass index (BMI, OR = 1.357 (95%CI 1.111-1.657)) and high density lipoprotein (HDL, OR = 0.774 (95%CI 0.705-0.850)) were the variables most significantly associated with the occurrence of metabolic syndrome in gout, but alcohol consumption did not show such associations. Gout is associated with the metabolic syndrome, and furthermore, obesity and dyslipidemia were the factors most associated with the syndrome in these patients. PMID- 16361818 TI - Transphyseal reconstruction of the anterior cruciate ligament using hamstring autograft in skeletally immature adolescents. AB - Eleven skeletally immature adolescents underwent anterior cruciate ligament reconstruction using a transphyseal tibial and femoral tunnel. An autologous quadrupled hamstring tendon was used in all cases and the average follow-up was 77.7 months. Clinical results were evaluated using Lysholm knee scores and a return to pre-injury sports activities. Radiological results were evaluated using side-to-side differences of instrumented laxities and growth disturbances compared with the uninjured side on final follow-up orthoroentgenograms. The mean Lysholm score was 97.8 (range 94-100) and mean side-to-side laxity difference was 2.4 mm (range 1-4). Ten of 11 patients returned to pre-injury sports activity. No patient had a leg length discrepancy of over 1 cm or a significant abnormal angular deformity of the knee joint. Therefore, anterior cruciate ligament reconstruction using the transphyseal tunnel and hamstring autograft in skeletally immature adolescents is believed to be a reliable treatment method, which is not associated with significant leg length discrepancy or abnormal angular deformity of the knee joint. PMID- 16361819 TI - The effects of intradermal vaccination with DNA encoding for the T-cell receptor on the induction of experimental autoimmune encephalomyelitis in B10.PL mice. AB - Intradermal gene administration was found to induce a more profound immune response than direct intramuscular gene injection. We performed intradermal vaccination of B10.PL mice with DNA encoding for the V 8.2 region of the T-cell receptors (TCR). Three weeks later, these mice were immunized with rat myelin basic protein (MBP). Daily mean clinical scores and mortality rate were lower in this group compared with controls. The proliferative responses of lymph node cells to rat MBP were slightly less in the vaccination groups than in the control groups (p < 0.05). However, we detected no differences between the two groups with regard to the production of MBP-specific IgG, IgG1, & IgG2a antibodies. The levels of cytokine mRNA expression in the vaccination groups were observed higher than in the control groups without antigen-specific stimulation, but all of cytokine expressions between the vaccination and control groups after antigen specific stimulation were identical. These results demonstrate that intradermal DNA vaccines encoding for TCR might prove to be useful in the control of autoimmune disease. PMID- 16361820 TI - Antitumor activity of TRAIL recombinant adenovirus in human malignant glioma cells. AB - Tumor necrosis factor-Related Apoptosis-Inducing Ligand (TRAIL) has been reported to specifically kill malignant cells but to be relatively nontoxic to normal cells. One of disadvantages to previous in vivo protocols was the need for large quantities of TRAIL recombinant protein to suppress tumor growth. To evaluate the antitumor activity and therapeutic value of the TRAIL gene, we constructed adenoviral vectors expressing the human TRAIL gene (Ad.hTRAIL) and transferred them into malignant glioma cells in vitro and tumors in vivo, as an alternative to recombinant soluble TRAIL protein. The results show that TRAIL-sensitive glioma cells infected Ad.hTRAIL undergo apoptosis through the production and expression of TRAIL protein. The in vitro transfer elicited apoptosis, as demonstrated by the quantification of viable or apoptotic cells and by the analysis of cleavage of poly (ADP-ribose) polymerase. Furthermore, in vivo administration of Ad.hTRAIL at the site of tumor implantation suppressed the outgrowth of human glioma xenografts in SCID mice. These results further define Ad.hTRAIL as an anti-tumor therapeutic and demonstrate its potential use as an alternative approach to treatment for malignant glioma. PMID- 16361821 TI - Natural menopause and risk of stroke in elderly women. AB - Although early natural menopause has been postulated to increase stroke risk, studies have not produced convincing results. We examined the associations between stroke risks and age at natural menopause or time since natural menopause. 5,731 naturally postmenopausal women more than 65 yr of age were followed from 1993 to 1998. Information on age at menopause and risk factors were obtained using mailed questionnaires. 186 cases of stroke occurred over a total 27,936 person-years. After adjusting for age, hypertension, and physical activity, age at menopause was not found to be significantly associated with stroke or cerebral infarction. However, adjusted relative risks (aRRs) showed a significant increasing tendency of hemorrhagic stroke versus age at menopause (aRRs, 0.66, 0.48, 1.00 and 2.33 for the following age groups at menopause; 40 44, 45-49, 50-54 [reference group], and > or =55 yr). Time since menopause (11 20, 21-30, and > or =31 yr) was not found to be significantly associated with cerebral infarction, or hemorrhagic stroke. Late menopause (menopause age > or =55 yr) showed a tendency of a lower risk of cerebral infarction (aRR, 0.79) and a higher risk of hemorrhagic stroke (aRR, 2.33). Further study is warranted to determine stroke risk in women during the decade following menopause. PMID- 16361822 TI - Intraabdominal cryptococcal lymphadenitis in a patient with systemic lupus erythematosus. AB - Cryptococcal infection is a rare, yet well recognized complication of systemic lupus erythematosus (SLE). We present a case of mesenteric and retroperitoneal cryptococcal lymphadenitis resulting in the obstruction of the stomach and proximal duodenum in a patient suffering from SLE, while recently she did not receive any immunosuppressive treatment. A 42-yr-old woman was admitted due to high fever and diffuse abdominal pain for three weeks. Abdominal computed tomography (CT) scan showed multiple conglomerated lymphadenopathies in the retroperitoneum and the mesentery resulting in luminal narrowing of the third portion of the duodenum. Cryptococcal lymphadenitis was proven by needle biopsy and she was treated with intravenous liposomal amphotericin B, followed by oral fluconazole. After fourteen-month antifungal therapies, the clinical symptoms and follow-up images improved. This case emphasize that the intrinsic immunological defects of SLE may be directly responsible for the predisposition to fungal infections. PMID- 16361823 TI - A case of primary gastric CD30-positive anaplastic large-cell lymphoma. AB - Gastric CD30-positive anaplastic large-cell lymphoma is a very rare disease. It is sometimes difficult to distinguish it from undifferentiated carcinoma, sarcoma and so on. We report here on a case of primary gastric anaplastic large-cell lymphoma. A 50-yr-old woman complained of epigastric pain and severe chest pain for 1 week. The gastroendoscopic examination revealed geographic mucosal irregularities with shallow ulceration at the antrum. She underwent a total gastrectomy. The gross finding of the resected stomach was an 8 x 4.5 cm sized ulceroinfiltrative lesion at the pyloric antrum along the lesser curvature. The microscopic examination revealed diffuse and solid proliferations of large atypical cells with pleomorphic nuclei. Immunohistochemically, the tumor cells were positive for CD30, vimentin and CD3, and this was a finding compatible with anaplastic large-cell lymphoma. To the best of our knowledge, this is the first such reported case in Korea. PMID- 16361824 TI - Familial interstitial lung disease in two young Korean sisters. AB - Most of the interstitial lung diseases are rare, chronic, progressive and fatal disorders, especially in familial form. The etiology of the majority of interstitial lung disease is still unknown. Host susceptibility, genetic and environmental factors may influence clinical expression of each disease. With familial interstitial lung diseases, mutations of surfactant protein B and surfactant protein C or other additional genetic mechanisms (e.g. mutation of the gene for ATP-binding cassette transporter A3) could be associated. We found a 21 month-old girl with respiratory symptoms, abnormal radiographic findings and abnormal open lung biopsy findings compatible with nonspecific interstitial pneumonitis that is similar to those of her older sister died from this disease. We performed genetic studies of the patient and her parents, but we could not find any mutation in our case. High-dose intravenous methylprednisolone and oral hydroxychloroquine were administered and she is still alive without progression during 21 months of follow-up. PMID- 16361825 TI - Infected infradiaphragmatic retroperitoneal extralobar pulmonary sequestration: a case report. AB - Infradiaphragmatic extralobar pulmonary sequestration is an extremely rare congenital malformation. It is more frequently diagnosed in the antenatal period due to routine ultrasonic examination of the fetus or in the first 6 months of life, though on rare occasions it is discovered incidentally in adults. A 32-yr old man presenting with epigastric discomfort and fever was referred. Computed tomographic scanning showed that a 16-cm, multiseptated, dumbbell-shaped, huge cystic tumor was located beneath the diaphragm. On the next day, 850 mL of thick yellowish pus was drained by sonography-guided fine needle aspiration for the purpose of infection control and diagnosis, but no microscopic organisms were found in repeated culture studies. Surgical removal of the cyst was performed through thoracoabdominal incision and most of these pathologic lesions were removed but we could not find the feeding arteries or any fistulous tract to surrounding structures. Histopathologic study revealed that it was extralobar pulmonary sequestration and culture study showed that many WBC and necrotic materials were found but there were no microorganisms in the cystic contents. We report the first case of an infected infradiaphragmatic retroperitoneal extralobar sequestration which was administered a staged management and achieved an excellent clinical course. PMID- 16361826 TI - A case of hypersensitivity pneumonitis caused by Penicillium species in a home environment. AB - We report a case of hypersensitivity pneumonitis in a 30-yr-old female housewife caused by Penicillium species found in her home environment. The patient was diagnosed according to history, chest radiograph, spirometry, high-resolution chest CT, and transbronchial lung biopsy. To identify the causative agent, cultured aeromolds were collected by the open-plate method. From the main fungi cultured, fungal antigens were prepared, and immunoblot analysis with the patient's serum and each fungal antigen was performed. A fungal colonies were isolated from the patient's home. Immunoblotting analysis with the patient's sera demonstrated a IgG-binding fractions to Penicillium species extract, while binding was not noted with control subject. This study indicates that the patient had hypersensitivity pneumonitis on exposure to Penicillium species in her home environment. PMID- 16361827 TI - Two novel mutations in the aquaporin 2 gene in a girl with congenital nephrogenic diabetes insipidus. AB - Congenital nephrogenic diabetes insipidus (CNDI) is a rare inherited disorder characterized by insensitivity of the kidney to the antidiuretic effect of vasopressin. There are three inheritance patterns of CNDI: the X-linked recessive form associated with vasopressin V2 receptor gene mutations, and the autosomal recessive and dominant forms associated with aquaporin-2 gene (AQP2) mutations. The evaluation for polyuria and polydipsia in a one-month-old Korean girl revealed no response to vasopressin and confirmed the diagnosis of CNDI. Because the child was female without family history of CNDI, her disease was thought to be an autosomal recessive form. We analyzed the AQP2 gene and detected a compound heterozygous missense point mutation: 70Ala (GCC) to Asp (GAC) in exon 1 inherited from her father and 187Arg (CGC) to His (CAC) in exon 3 inherited from her mother. The first mutation is located within the first NPA motif of the AQP2 molecule and the second one right after the second NPA motif. This is the first report to characterize AQP2 mutations in Korean patients with autosomal recessive CNDI, and expands the spectrum of AQP2 mutations by reporting two novel mutation, 70Ala (GCC) to Asp (GAC) and 187Arg (CGC) to His (CAC). PMID- 16361828 TI - Spontaneous uterine rupture in the first trimester: a case report. AB - Uterine rupture is one of the most feared obstetric complications affecting the pregnant woman and fetus. Most of the cases have various risk factors and mainly occur during the second or third trimester. However, spontaneous uterine rupture during the first trimester is extremely rare. We experienced a case of spontaneous uterine rupture in a 36-yr-old multiparous woman without definite risk factors. The initial impression was a hemoperitoneum of an unknown origin with normal early pregnancy. Intensive surgical method would be needed for accurate diagnosis and immediate management in bad situation by hemoperitoneum even though a patient was early pregnancy. PMID- 16361829 TI - An infantile case of Sturge-Weber syndrome in association with Klippel-Trenaunay Weber syndrome and phakomatosis pigmentovascularis. AB - Sturge-Weber syndrome can be associated with facial port-wine stains and intracranial calcification, and concurrent Klippel-Trenaunay-Weber syndrome has been reported. Klippel-Trenaunay-Weber syndrome is a rare congenital mesodermal phakomatosis characterized by cutaneous hemangiomas, venous varicosities and soft tissue or bone hypertrophy of the affected extremities. This report is presented a rare case of the Sturge-Weber syndrome in combination with the Klippel Trennaunay syndrome and phakomatosis pigmentovascularis in a 4-month-old infant. He showed nevus flameus on the right leg and both part of the face and back, leptomeningeal angiomatosis on right hemisphere, hypertrophy of the right leg, hemiconvulsion on the left and also evidences of congenital glaucoma and nevus of Ota. Very rare case combined with these three kinds of phakomatosis has been reported. PMID- 16361830 TI - A catastrophic-onset longitudinal myelitis accompanied by bilateral internuclear ophthalmoplegia in a patient with systemic lupus erythematosus. AB - Transverse myelitis (TM) extending from midbrain to the entire spinal cord accompanied by internuclear ophthalmoplegia is extremely rare but cause serious central nervous system complications in patients with systemic lupus erythematosus. We report a case of a 28-yr-old woman with TM extending from the midbrain to the conus medullaris longitudinally and internuclear ophthalmoplegia associated with systemic lupus erythematosus. Her neurological symptoms had an abrupt catastrophic onset and rapidly progressed to respiratory failure within 24 hr. Bilateral internuclear ophthalmoplegia was also followed by TM. Brain MR images showed definite brainstem lesions, which were deeply associated with internuclear ophthalmoplegia, and diffuse signal changes in the whole spinal cord, medulla, pons and midbrain. Clinical improvement of her ophthalmoplegia and of neurological dysfunction of the upper extremities was noted after prompt and aggressive treatment with intravenous pulsed methylprednisolone and cyclophosphamide. However, the neurological dysfunction of the lower limbs and bladder and colon paralysis were almost unchanged until six months passed. PMID- 16361831 TI - GSTM1, GSTT1 and GSTP1 polymorphisms in the Korean population. AB - The isoenzymes of the glutathione s transferase (GST) family play a vital role in phase II of biotransformation of many substances. Using a multiplex polymerase chain reaction and a direct sequencing analysis, the frequencies of GSTM1, GSTT1, and GSTP1 polymorphisms were evaluated in 1,051 Korean male subjects. We found that 53.8% of the individuals had the GSTM1 null genotype and 54.3% had the GSTT 1 null genotype. The genotypic distribution of GSTP1 was Ile105/Ile105 in 68.4%, Ile105/Val105 in 29.1% and Val105/Va105 in 2.5%. The most frequently observed combination of GSTM1, GSTP1 and GSTT1 genotypes was Null type/Ile105/Ile105/Null type, while the combination of Non-null type/Val105/Val105/Non-Null type was not observed. We found that the genotype distributions of three GST isoenzymes in the Koreans are similar to those reported in Asians and previously reported Koreans. We believe our results, which are represented by a large population, are reliable estimates of the frequencies of the polymorphic GST alleles in the Koreans and will help future researches on GST polymorphisms. PMID- 16361833 TI - Dialyzer and machine technologies: application of recent advances to clinical practice. AB - Although hemodialysis is a mature therapy, a growing population of patients with more complex medical problems and limitations on resources will require technological innovations to improve the safety, reliability and efficiency of the therapy. The past several years have seen design changes to dialyzers that have provided incremental improvements in small solute clearance and more substantial improvements in the clearance of large solutes. New functions have been added to dialysis machines that help ensure reliable delivery of the dialysis prescription and enable full advantage to be taken of improvements in dialyzer clearance of large molecules. In addition, feedback control systems have been developed that may help reduce the untoward side effects which many patients experience during hemodialysis. Whether or not a particular innovation enters routine clinical use will depend on demonstrating that it improves clinical outcomes, its cost, and, in some cases, on a more enlightened approach by regulatory authorities. PMID- 16361834 TI - The importance of water quality and haemodialysis fluid composition. AB - Treatment of renal failure by haemodialysis uses dialysis fluid to facilitate the normalization of electrolyte and acid base abnormalities and the removal of low molecular weight uraemic compounds present in the plasma such as urea. The dialysis fluid is a continuously produced blend of treated tap water and a concentrated solution containing electrolytes, buffer, and glucose. The water used originates as drinking water but undergoes additional treatment. Recent surveys have indicated that the chemical and microbiological content of such water frequently fails to meet the requirements of established standards, and its bacterial content arising from the presence of a biofilm in the water distribution network or the hydraulic circuit of the dialysis machine is a contributory factor to the chronic inflammatory state in patients undergoing regular dialysis. The composition of the dialysis fluid plays an important role in the modulation of complications associated with end-stage renal disease, as well as those associated with the treatment itself. The avoidance of complications arising from water contaminants requires a constant and vigorous attention to water quality, whilst with the composition of electrolytes and buffer there is a trend towards greater individualization to provide a high degree of treatment tolerance. PMID- 16361835 TI - Patient safety in the dialysis facility. AB - Medical mistakes that harm patients have occurred in dialysis facilities, just as they have been reported to cause death in hospitals. A plan to recognize and prevent mistakes includes several elements. (1) Establish a culture of safety, where safe practices are a priority and reporting of adverse events and 'near misses' is encouraged in a non-punitive environment. (2) Redesign processes of care to minimize patient falls, medication errors, bleeding from vascular accesses, dialyzer reuse errors and other problems. (3) Perform root cause analyses of adverse events and 'near misses'. (4) Involve patients in safety efforts, including maintaining an accurate, up-to-date medication list. (5) Dialysis facilities should function as high-reliability organizations, focusing on the possibility of failures, expecting to make errors, training to recognize and recover from them and making system reforms following root cause analyses. PMID- 16361836 TI - Establishing a successful home dialysis program. AB - The renewed interest in home dialysis therapies makes it pertinent to address the essentials of establishing and running a successful home dialysis program. The success of a home program depends on a clear understanding of the structure of the home program team, the physical plant, educational tool requirements, reimbursement sources and a business plan. A good command of the technical and economic aspects is important, but the primary drivers for the creation and growth of a home dialysis program are the confidence and commitment of the nephrological team. PMID- 16361837 TI - Payment for performance: in sickness and in health. For better or for worse? AB - American consumers and payers are increasingly questioning the value of health care as costs increase amid reports of poor quality. Rewarding health care providers who apply evidence-based, clinical practice guidelines (CPGs) to obtain desired patient outcomes seems an intuitive, innovative method to improve health care value. Although there are many unanswered questions about payment for performance (P4P), Congress and the Centers for Medicare and Medicaid Services are developing a rapid timetable to begin P4P. Many private payers have already begun P4P programs. Currently, we lack randomized, controlled studies proving the safety, effectiveness, and patient centeredness of P4P. Many see publicly reporting provider performance as a surrogate for P4P. There is a growing literature documenting the effect of publicly reporting provider performance on the medical community and patient outcomes. It changes provider behavior, but occasionally, in undesirable ways (for example, adverse risk selection, or 'cherry picking'). Publicly reporting or rewarding provider performance based on applying evidence-based CPGs may also have unforeseen negative patient outcomes. P4P should proceed carefully while considering unintended consequences for patients and providers. P4P should be pilot tested in the proper target patient and provider populations. Thus, we need to study how CPGs interact with one another in specific patient populations. The updating process for CPGs should include evidence-based statements concerning their impact on real patients with multiple chronic illnesses. PMID- 16361838 TI - What is uremia? Retention versus oxidation. AB - The uremic syndrome is characterized by the retention of a host of compounds which in healthy subjects are secreted into the urine by the healthy kidneys. These compounds disturb many physiologic functions, resulting in toxicity. However, many of the retained compounds as well as many of the pathophysiologic actions of the known retention solutes remain unknown. In this publication, we review recent information on uremic toxicity. Especially the difficulty to remove compounds, such as protein-bound compounds and larger molecules, seems to play a crucial role. New strategies enhancing their removal might be highly useful. Part of the retained compounds are the result of oxidative processes due to the inflammatory status of uremic patients; however, other compounds are not, and even the concentration of oxidative compounds will be further increased by disturbances of urinary clearance. PMID- 16361839 TI - Dialysis patients and cardiovascular problems: Can technology help solve the complex equation? AB - Patients with end-stage kidney disease undergoing chronic hemodialysis (HD) present higher mortality rates compared with the general population. Once patients are on HD, the risk of cardiovascular death is approximately 30 times higher than in the general population and remains 10-20 times higher after stratification for age, gender, and the presence of diabetes. About half the deaths of patients on dialysis are attributed to cardiovascular causes including coronary heart disease, cerebrovascular disease, peripheral vascular disease, and heart failure. The cardiovascular burden of the HD patient arises from three different sources: risks inherent to the patient and the uremic syndrome, traditional risk factors, and risk factors related to the dialysis therapy. Based on these considerations and the fact that several aspects of the dialysis procedure can either add to the cardiovascular burden or modify the existing burden, new technologies should be directed towards the approach of a potential 'cardioprotective dialysis therapy'; such an approach may be facilitated by the application of new techniques and advanced dialysis machines. Created to make dialysis easy and safe, new machines feature several options that make patient monitoring and online hemodiafiltration therapy routine procedures. These and other features will possibly make dialysis better tolerated and more efficient in protecting patients from undesirable or potentially fatal cardiovascular events. PMID- 16361840 TI - Kidneys, hearts, hormones and immunomodulators: integrated understandings. AB - The biological processes governing vascular endothelial cell health are complex and highly redundant. The mechanisms involved in cell injury and repair remain an area of intense study. This paper attempts to review current knowledge regarding hormones such as vitamin D, erythropoietin and aldosterone within the context of what we understand of the inflammatory cascade. This is not a comprehensive review of vascular biology, and many important details and background information have been omitted due to space and complexity considerations. The integration of exocrine and endocrine functions within the kidney, the intimate relationship of those functions with vascular health, and the clinical observations of high prevalence of cardiovascular disease in chronic kidney disease warrant a better understanding of this area by clinicians and basic researchers. PMID- 16361841 TI - Association between inflammation and malnutrition as risk factors of cardiovascular disease. AB - Cardiovascular disease is the leading cause of death among dialysis patients. The relative risk of mortality increases as serum albumin concentration and body mass index decline. While these are generally associated with nutritional status, inflammation causes sarcopenia and decreased albumin concentration by reducing synthesis of proteins and increasing their catabolic rate. While inflammation can arise from atherosclerotic blood vessels, systemic inflammation from any source can alter the vascular endothelium and plasma protein composition in ways that promotes vascular injury. High-density lipoprotein synthesis is decreased and the high-density lipoprotein present is less capable of reducing inflammation. Activation of neutrophils favors lipoprotein oxidation. Surprisingly, while obesity is associated with cytokine production in patients without renal failure, as well as among dialysis patients, increased body mass index, whether reflecting muscle mass or adipose tissue, is associated with a decline in mortality rates. PMID- 16361842 TI - Vascular calcification in patients with chronic kidney disease. AB - Chronic kidney disease (CKD) represents an extremely common condition, and cardiovascular diseases are frequently reported in this patient population. Traditional risk factors are not accurate prognostic predictors in CKD patients, and new potential markers to predict the cardiovascular involvement in uremic patients need to be identified. Vascular calcification (VC) represents a hallmark of the atherosclerotic process in CKD. This review summarizes the processes responsible for VC (particularly focusing on the mechanisms operative in the presence of renal dysfunction), discusses the utility of computer tomography modalities in the detection of VC in patients with CKD, and reports the potential role of VC as pathophysiological link between kidney disease and cardiovascular events. PMID- 16361843 TI - Salt--a potential 'uremic toxin'? AB - It has been known for decades that salt (NaCl) determines extracellular volume as well as blood pressure and is one cause of hypertension. The difficulty to control the NaCl balance and thus treat sodium overload and hypertension in patients on dialysis has been recognized by Scribner in the early days of dialysis. In recent years, an impressive body of evidence has accumulated indicating that in essential hypertension, NaCl--blood pressure independently- causes target organ damage such as left ventricular hypertrophy, microalbuminuria, and increased aortic stiffness. It has further been recognized that NaCl increases oxidative stress and, again blood pressure independently, amplifies tissue injury induced by aldosterone. In renal damage models, progression is dramatically accelerated by high NaCl intake. Sodium as a potential culprit in progression to target organ damage in terminal renal failure has not been well investigated so far. However, it is possible, and indeed likely, that sodium plays an adverse role in the genesis of target organ damage in terminal renal failure. PMID- 16361844 TI - Uric acid--a uremic toxin? AB - Uric acid might often be regarded as a simple marker of renal disease. Although it is well known that hyperuricemia causes gout which is associated with renal insufficiency and cardiovascular disease, one might think that it could attribute to the intrarenal urate crystal, but not to uric acid per se. In order to clarify the role of uric acid in the kidney, we hypothesized that uric acid causes renal disease. To generate mild hyperuricemia without intrarenal crystal in rats, we used low doses of an uricase inhibitor (2% oxonic acid). Hyperuricemia induced systemic hypertension, glomerular hypertrophy/hypertension, afferent arteriolar sclerosis, and macrophage infiltration in normal rat kidney. In progressive renal disease, such as cyclosporine nephropathy and remnant kidney in rat, uric acid accelerated the progression of renal disease. Thus, we concluded that uric acid is not a simple marker, but a cause of renal disease. PMID- 16361845 TI - A critical assessment of uremia research. AB - There are considerably fewer randomized controlled trials investigating hemodialysis (HD) than other fields of internal medicine, and no significant improvements have been observed over time. Only the National Cooperative Dialysis Study and the HEMO trial were based on hard endpoints such as morbidity and mortality, but neither considered on-line hemodiafiltration or super-flux membranes, which are thought to provide a number of advantages in terms of the cardiovascular condition of uremic patients. However, results of well-designed clinical trials showing that increasing convection may improve the clinical outcome of HD patients are still lacking. The need for maximizing removal of uremic toxins calls for more frequent HD sessions, but this may be affected by many organizational problems. Therefore, well-designed, long-term clinical trials are urgently needed to investigate which currently available therapeutic instruments can improve the clinical outcome of uremic patients. PMID- 16361846 TI - Access monitoring is worthwhile and valuable. AB - During the past several years, a limited number of small clinical trials have questioned the role of surveillance in the management of vascular accesses, since the prolongation of access longevity until replacement was not altered. Although prolongation of access life span is an important endpoint, it is not the only one. Reduction in thrombotic events reduces the risks to the patient resulting from loss of access patency. The body of evidence suggests that the detection of stenosis and prevention of thrombosis are valuable. When a test indicates the likely presence of a stenosis, venography or fistulography should be used to definitely establish the presence and the degree of the stenosis. In most cases, angioplasty should be performed if the stenosis is greater than 50% by diameter. The value of routine use of any surveillance technique for detecting anatomic stenosis alone without concomitant functional assessment by measurement of access flow, venous pressure, recirculation, or other physiologic parameter has not been established. Stenotic lesions should not be repaired merely because they are present. If such correction is performed, then intra-procedural studies of access flow or intra-access pressure prior to and following percutaneous transluminal angioplasty should be conducted to demonstrate a functional improvement with a 'successful' percutaneous transluminal angioplasty. PMID- 16361847 TI - Strategies to minimize tunneled hemodialysis catheter use. AB - While the use of arteriovenous grafts has recently declined, there has been an astronomical increase in hemodialysis patients dialyzing with tunneled dialysis catheters (TDCs). Recent data have indicated that over 70% of the patients with end-stage renal disease initiate dialysis with a catheter. Additionally, up to 27% of the end-stage renal disease patients in the US are using TDCs as their permanent access, with placement rates having doubled since 1996. Although most modern catheters claim to provide adequate blood flow for dialysis, they are associated with the highest incidence of complications, morbidity and mortality when compared with other types of vascular access. It is for these reasons that the National Kidney Foundation Dialysis Outcomes Quality Initiative guideline 30 as well as the Fistula First Change Concept 7 emphasize limiting the use of catheters and fostering the creation of arteriovenous fistulae. Early referral has clearly been shown to minimize the use of TDCs and maximize fistulae. This report focuses on the role of additional measures that minimize TDC use, such as dialysis modality presentation and peritoneal dialysis, vascular access education, preoperative vascular mapping and salvage of early failure and thrombosed fistulae. PMID- 16361848 TI - Cause and consequences of sympathetic hyperactivity in chronic kidney disease. AB - Patients with chronic kidney disease and patients undergoing hemodialysis treatment show a sustained overactivity of the sympathetic nervous system, which originates from signals arising in the failing kidneys and traveling via afferent renal nerves to cardiovascular centers in the brainstem. Additional important factors are increased levels of angiotensin II and asymmetrical dimethylarginine. The sympathetic overactivity contributes to hypertension and cardiovascular morbidity and mortality in that patient population. Sympathetic overactivity can be reduced by adrenergic receptor blockers, centrally acting sympathicolytic drugs such as moxonidine and rilmenidine, angiotensin-converting enzyme inhibition, and angiotensin II type 1 receptor antagonists. Daily short hemodialysis and long nocturnal hemodialysis may reduce the elevated sympathetic activity, possibly because of an increased clearance of asymmetrical dimethylarginine, an endogenous nitric oxide synthase inhibitor. Prospective trials examining the potential impact of both beta-blockers and centrally acting sympatholytic drugs on cardiovascular mortality in chronic kidney disease and hemodialysis patients are very much needed. PMID- 16361849 TI - Proven strategies to reduce cardiovascular mortality in hemodialysis patients. AB - BACKGROUND: In hemodialysis patients, left ventricular hypertrophy (LVH) correlates with mortality. The reason for LVH in uremics is multifactorial. The primary objective of our study was to investigate the effects of a multi interventional treatment strategy on LVH. METHODS: In 230 ambulatory patients, including patients with coronary artery disease, diabetes, diastolic and systolic dysfunction, we continued optimized cardiac therapy (beta-blockers, angiotensin converting enzyme inhibitors and angiotensin receptor blockers) with full anemia correction by intravenous epoetin-beta. The dose of epoetin-beta for maintaining target hemoglobin (Hb) was 68 +/- 23 IU/kg/week. Serial echocardiograms were recorded every 3-6 months. The mean observation period was 4.8 +/- 1.2 years. RESULTS: Mean Hb at baseline was 11.2 +/- 2.0 versus 14.1 +/- 1.4 g/dl (p < 0.001) at study end. There was a significant reduction in left ventricular mass index (LVMI: 159 +/- 50.4 vs. 130.2 +/- 42.7 g/m(2); p < 0.001). In a subgroup of 2/3 of the patients, LVMI returned to normal (169 +/- 33 vs. 114 +/- 14 g/m2; p < 0.001). CONCLUSION: Baseline LVMI (p < 0.001), Hb increase (p < 0.03), and triple cardiac therapy (p < 0.03) were significant and independent prognostic factors for a reduction in LVMI. The annual cardiovascular mortality was 5%. Even anemia correction from 12 to 14 g/dl results in further (p < 0.001) regression of LVMI. PMID- 16361850 TI - Peritoneal dialysis in Europe: an analysis of its rise and fall. AB - There is a wide variability in the utilization of peritoneal dialysis throughout Europe that cannot be explained by differences in medical factors alone. Over the last years, the utilization of peritoneal dialysis has even changed in individual countries. The main factors of this variation are not medical, but rather changes in reimbursement and other financial considerations. The overall structure of the health care in an individual country is to a large extent determining the use of a renal replacement therapy modality. PMID- 16361851 TI - Chronic kidney disease management--what can we learn from South African and Australian efforts? AB - BACKGROUND: The prevalence of chronic kidney disease is on the rise. Our objective is to describe two programs to improve the awareness and management of hypertension, renal disease, and diabetes in remote Australian Aboriginal and urban and periurban South African communities. We focus on how the Australian Aboriginal and South African Chronic Disease Outreach Programs have worked together. METHODS: The establishment of prevention programs in developing countries is a challenge. The paper evaluates these challenges, including accessing international aid. The programs advocate that regular integrated checks for chronic disease and their risk factors are essential elements of regular adult health care. Programs should be run by primary health workers, following algorithms for testing and treatment, and a backup provided by nurse coordinators. Constant evaluation is essential to develop community health profiles and adapt program structure. RESULTS: Both programs are discussed, including how they are organized to deliver preventative and treatment strategies. The challenges and adaptations required are outlined. CONCLUSIONS: It is the aim of the international kidney community to prevent chronic kidney disease. The South African and Australian groups highlight the need for a systematic and sustained approach to the management of chronic diseases to achieve this goal. PMID- 16361852 TI - Randomized trials of frequent hemodialysis--infinite possibilities. AB - Current understanding of the benefits of daily and nocturnal hemodialysis is limited by the designs used in previous studies which tested these therapies. There is increasing momentum to evaluate these therapies in randomized controlled trials. By establishing the efficacy, safety and delivery of daily and nocturnal hemodialysis, such trials will guide policy and funding decisions regarding the role of these therapies in the treatment of end-stage renal disease. However, in designing trials that fulfill these goals, many questions arise. What study population should be used? How should the intervention be defined? Which outcomes should be evaluated? Several methodological issues particular to trials of frequent dialysis require additional consideration. The 'ideal' trial in terms of methods must be balanced against the feasibility of implementation and budgetary constraints. Here, we explore some of these issues. PMID- 16361853 TI - Debate forum: levocarnitine therapy is rational and justified in selected dialysis patients. AB - Carnitine is a metabolic cofactor which is essential for normal fatty acid metabolism. Patients with chronic kidney disease on dialysis have been shown both to suffer from disordered fatty acid metabolism and to have a significant deficiency in plasma and tissue carnitine. Aberrant fatty acid metabolism has been associated with a number of cellular abnormalities such as increased mitochondrial permeability (a promoter of apoptosis), insulin resistance, and enhanced generation of free radicals. These cellular abnormalities have, in turn, been correlated with pathological clinical conditions common in dialysis patients including cardiomyopathy with attendant hypotension and resistance to the therapeutic effect of recombinant human erythropoietin (EPO). In 1999, the Food and Drug Administration approved levocarnitine injection for the prevention and treatment of carnitine deficiency in patients on dialysis based on documentation of free plasma carnitine levels in dialysis patients similar to other serious carnitine deficiency states for which treatment was required. Data analysis performed by expert panels convened by both the American Association of Kidney Patients and, subsequently, the National Kidney Foundation recommended a trial of levocarnitine therapy for specific subsets of dialysis patients including those with EPO resistance, dialysis-related hypotension, cardiomyopathy and muscle weakness. In 2003, the Centers for Medicare and Medicaid services convened a Medical Advisory Committee which established reimbursement on a national level for carnitine-deficient dialysis patients who had either dialysis-related hypotension or EPO resistance. Recently, a correlation between reductions in hospitalization rates of dialysis patients receiving levocarnitine therapy has been demonstrated in a large retrospective study. Despite data-based recommendations and national reimbursement, only a small minority of dialysis patients have been prescribed a therapeutic trial of levocarnitine. Whereas the reasons for the reluctance of nephrologists to prescribe this therapeutic trial are unclear, possible explanations include a lack of appreciation of the pivotal role played by carnitine in cellular metabolism and the strength of evidence for a substantial deficiency of carnitine in dialysis patients, an underestimation of the prognostic import of EPO resistance and dialysis-related hypotension, inadequate dissemination of the clinical trial data supporting the use of levocarnitine in dialysis patients, and the heterogeneous clinical response of dialysis patients to levocarnitine therapy. Difficulties in documenting both initial eligibility and evidence of improvement as a result of therapy may also be a contributing factor. This paper discusses the biological role of carnitine and its particular relevance to dialysis patients. Clinical trial data concerning an effect of therapy on EPO resistance and dialysis-related hypotension are summarized along with a discussion of the logic behind the use of levocarnitine in dialysis. Finally, the difficulties posed by a reimbursement policy based on clinical as opposed to laboratory endpoints and a heterogeneous response to therapy are addressed. PMID- 16361854 TI - Debate forum: carnitine supplements have not been demonstrated as effective in patients on long-term dialysis therapy. AB - The database for carnitine supplements in dialysis includes no large-scale randomized trials and no registered trials. Medical practitioners prefer to make treatment decisions based on the outcome of randomized clinical trials, with appropriate controls. Furthermore, registered trials provide a further level of integrity, since trial registration avoids publication bias by ensuring that all outcomes are reported, including trials that are not completed. Positive effects reported from carnitine administration in dialysis patients include decreased erythropoietin dose, increased hematocrit, less intradialytic hypotension, and less fatigue. The evidence for carnitine effectiveness is limited to trials that are mostly open-label and that include no more than a total of 1,000 patients. An analysis of recent carnitine administrations to patients in a large dialysis practice database indicates no overall change in hemoglobin or erythropoietin dose following 6 months of carnitine administration. As outcomes of controlled trials with appropriate power to examine for the benefits of carnitine are not yet available, the dialysis practitioner cannot justify the administration of carnitine. PMID- 16361855 TI - A nexus of progression of chronic kidney disease: charcoal, tryptophan and profibrotic cytokines. AB - Fibrosis plays a role in the pathogenesis of progressive chronic kidney disease (CKD). The inhibition of the renin-angiotensin system, which promotes fibrosis, has become the standard of care in the treatment of patients with CKD. A novel charcoal compound, AST-120, has been used for over a decade in Japan to prevent progression of CKD. It is thought that the oral administration of AST-120 blocks the intestinal absorption of tryptophan-derived indole. This prevents the hepatic conversion of indole to indoxyl sulfate (IS). IS has been shown to stimulate the production of profibrotic cytokines such as transforming growth factor-beta. AST 120 lowers IS in a dose dependent fashion and does not change the creatinine appearance rate in the urine. Enteric capsules containing Bifidobacterium longum have been shown to prevent progression of CKD in a preliminary study. These findings suggest that prospective clinical trials be undertaken to determine if these other potential methods of inhibiting fibrosis are useful in slowing progressive CKD. PMID- 16361857 TI - The education of speech-language pathologists in the United States of America. AB - The education of speech-language pathologists and audiologists is provided by over 250 colleges and universities at the master's and doctoral level. The educational programs for speech-language pathologists and audiologists consist of academic coursework, clinical education, and supervised experience. Coursework and clinical education include study in liberal arts and science and human communication in addition to study in all aspects of communication sciences and disorders, including cultural and linguistic diversity. The programs may take 2 to 3 years beyond the bachelor's degree. Audiologists will require a doctoral degree beginning in 2012. Educational programs are accredited by the Council on Academic Accreditation of the American Speech-Language Hearing Association. Individuals who wish so earn the Certificate of Clinical Competence from the American Speech-Language Hearing Association. PMID- 16361856 TI - Oxidant and carbonyl stress-related apoptosis in end-stage kidney disease: impact of membrane flux. AB - Apoptosis is a highly regulated process which mostly affects cell-mediated immunity. In this open-label, randomized, prospective clinical study, we determined the impact in 10 hemodialysis patients treated with high-, medium-, and low-flux membranes on spontaneous or plasma-induced apoptosis, on monocytes, as well as on oxidant and carbonyl stress. High- and medium-flux membranes significantly reduced patients' plasma-dependent proapoptotic activity on U937 monocytic cell lines. Patients who had the highest levels of plasma-induced proapoptotic activity exhibited the highest plasma levels of advanced oxidation protein products (AOPPs) and carbonyls. Plasma carbonyl residues but not AOPPs were significantly lowered. Finally, a significant correlation could be drawn between the extent of plasma-induced proapoptotic activity and both plasma carbonyl and AOPP levels. PMID- 16361858 TI - Education of speech-language pathologists and audiologists in Brazil. AB - The field of speech-language pathology (SLP) in Brazil, named 'fonoaudiologia', comprises both a therapeutic approach to communication disorders and audiology and was officially recognized on December 9, 1981 (law No. 6965). University programs exist since the 1960s. The undergraduate level is a 4-year honors Bachelor of Science program and requires at least a 3,700 h of coursework. Since 1996 four areas of specialization were established: language, audiology, voice and oral myology, requiring a minimum of 500 h of course. Graduate programs in the narrower sense,master's degree and doctorate, exist since the 1970s. Brazil is a 180-million inhabitant country with approximately 25,000 speech-language pathologists, of which 2,700 are specialists, 800 masters and 210 doctors. There are almost 100 undergraduate programs and 70 specialization courses; however, for master's degree and doctorate purposes there are only 8. PMID- 16361859 TI - Competency-based occupational standards: influences on Australian speech pathology education. AB - Speech Pathology Australia developed competency-based occupational standards in response to a national government initiative across all professions and these standards have formed the basis for the accreditation process of speech pathology education programmes in Australia since 1994. This paper presents a critical appraisal of the contribution of a competency framework to curricular design, focussing on both content coverage and teaching methodologies. Also, the assessment of learning promoted through a competency-based framework is considered in relation to models of critical thinking. PMID- 16361860 TI - US doctoral education: critical shortages and plans for reshaping the future. AB - The paper describes the major features of doctoral curricula in the United States for speech-language pathology, speech, language, hearing science and the AUD degree in audiology. Critical shortages in doctoral personnel in the United States, factors that are considered to be contributing to the problem of shortages and actions that have been recommended for reshaping the future of doctoral education are also discussed. PMID- 16361861 TI - Education in logopaedics in the Maltese Islands. AB - The Communication Therapy (CT) Division of the University of Malta is relatively small and young and is the only local training establishment of speech-language pathologists (SLPs) in the Maltese Islands. It aims to achieve a distinctive profile of excellence and international standing in the education and training of caring, accountable and autonomous SLPs. Various strategies are adopted by the CT Division in order to reach its goals and to overcome its limitations and challenges. These include running its own Teaching and Research Clinic, engaging in European staff/student exchange and conducting research projects with other organizations to develop scientific data and tools for use with the Maltese communication-disordered population. PMID- 16361862 TI - Issues of Japanese speech therapy education considering the 1995 Guidelines of International Association of Logopedics and Phoniatrics (IALP). AB - The historical background of the educational system for Japanese speech and language therapists (pathologists) and the challenge facing us today are reviewed. It took nearly 30 years for speech therapists and physicians to come to a consensus in defining the status of speech therapists (pathologists) and their educational curriculum before the 1997 national law for Japanese speech therapists was passed. Despite the formulation of the International Association of Logopedics and Phoniatrics (IALP) Education Guidelines of 1995, little attention was paid to incorporate it into the 1997 national law. This paper discusses some challenges in the area of speech therapists' status and education system that exist today. PMID- 16361863 TI - Education for practice in the UK and Ireland: implementing problem-based learning. AB - OBJECTIVE: This paper outlines the education of speech and language therapists in the UK and Ireland, and presents a preliminary study of student therapists' perceptions of problem-based learning (PBL) as a learning strategy in preparation for clinical work. PBL has been used extensively in medical and dental education in Europe, in Canada, and in the Middle East, and has been applied to speech and language therapist education in Sweden and in Australia. Its implementation in the UK and Ireland is relatively new. METHODS: A survey questionnaire was circulated to students in two centres via e-mail. Questions posed included student impressions of the most and least useful elements of PBL in their preparation for clinical practice, as well as how they considered improvements could be made; student reflections regarding PBL were also sought. RESULTS AND CONCLUSIONS: Findings support the implementation of PBL in the education of speech and language therapists, with more experienced students showing more positive support for PBL. Issues raised by the study include emphasis on clinical relevance of problems, particularly in the early years of the course. The majority of students regarded PBL as directly relevant for clinical preparation. PMID- 16361864 TI - Preparation of speech-language pathologists in the United States: The Master's degree. AB - In order to become credentialed as a speech-language pathologist in the United States, a masters's degree is required. This degree integrates course work with clinical practica. Course work covers normal anatomy and physiology involved in human communication and swallowing, disorders affecting these functions and methods of evaluation and treatment across the life span. Generally this master's degree education requires 1(1/2) to 2 years for students with some background course work on the undergraduate level. PMID- 16361865 TI - Education of speech-language pathologists around the world: The Scandinavian experience. AB - The object of this paper is to give an overview of the education and training programmes in logopedics offered in the Scandinavian/Nordic countries and to provide some information about the development of the programmes in Scandinavia since their foundation in the 1920s. In order to collect information about the various programmes, a questionnaire comprising 12 questions was sent to representatives for education programmes and national societies affiliated to the IALP. In the past all the countries used to provide possibilities for teachers to continue their education with courses in logopedics to become 'special teachers'. These programmes no longer exist except in Norway. Degree programmes at bachelor's and/or master's level are now offered in all the Nordic countries. It can be concluded that the education and training in logopedics have developed into university degree programmes, with few exceptions, in the Nordic countries. PMID- 16361866 TI - Expression of vascular adhesion protein-1 in atopic eczema. AB - BACKGROUND: Vascular adhesion protein-1 (VAP-1) is an adhesion molecule with an enzymatic activity which partakes in the migration process of lymphocytes. The aim of this study was to investigate VAP-1 expression in atopic eczema (AE) in comparison with healthy controls and psoriatic individuals. MATERIAL AND METHODS: Forty adult patients suffering from AE aged between 18 and 56 years were included in the study. The control group consisted of 35 healthy volunteers aged between 19 and 49 years and of 71 psoriatic patients aged between 23 and 89 years. The serum concentration of soluble VAP-1 (sVAP-1) was evaluated by ELISA and VAP-1 expression in the skin by immunohistochemistry. RESULTS: Serum level of sVAP-1 in AE patients before treatment was significantly higher compared with healthy volunteers. Similarly, a higher mean number of VAP-1-positive vessels was found in both lesional and nonlesional atopic skin compared with healthy skin. Treatment of AE resulted in a significant reduction in the serum level of sVAP-1. On the other hand, both the serum level of sVAP-1 and the number of dermal vessels with expression of VAP-1 were significantly lower in AE patients compared with psoriatic individuals. CONCLUSION: This study indicates the important role of VAP-1 in the pathogenesis of chronic inflammatory cutaneous disorders, including AE. PMID- 16361867 TI - Changes in localization of amino acids in the detached cat retina. AB - PURPOSE: To investigate the distribution of amino acids (glutamate, aspartate, glutamine, GABA, glycine) in detached retinas with minimum postmortem artifact and to clarify the relation between amino acid distribution and histopathological change in the outer portion of detached retinas. METHODS: Unilateral retinal detachment was produced in cats by injecting 0.25% sodium hyaluronate into the subretinal space using a glass micropipet. The eyes were fixed by perfusion for 10 min, 1, 3, 6 and 24 h, 2, 3 and 7 days after detachment and then examined under conventional light- and electron-microscopic immunocytochemistry. RESULTS: For glutamate, aspartate and glutamine, the inner segments and perikarya of the photoreceptor cells, which were not immunopositive in the normal retinas, showed various degrees of immunoreactivity immediately after retinal detachment. Photoreceptor cells with the strong immunoreactivity developed necrosis. The staining pattern of GABA and glycine scarcely changed during the course of retinal detachment. CONCLUSIONS: Excess intracellular glutamate, aspartate and glutamine in photoreceptor cells may cause a part of neuronal death after retinal detachment. PMID- 16361868 TI - A different medicine is possible. PMID- 16361869 TI - Reflections on relevance: Psychotherapy and Psychosomatics in 2005. AB - This article focuses on the mission and relevance of articles published in Psychotherapy and Psychosomatics in 2005. Four areas of articles relevant to the mission of Psychotherapy and Psychosomatics are reviewed: (1) new (psycho )therapies; (2) cognitive-behavioral therapy; (3) new or newly defined psychopathology, and (4) the impact of or impairment due to various illnesses or treatments. In addition, other clinically important and relevant issues, such as the need for new treatments, suicide during psychotherapy, technological innovations used in psychotherapies and methodology of psychotherapy trials are discussed. PMID- 16361870 TI - A systematic review of treatments for electromagnetic hypersensitivity. AB - BACKGROUND: Electromagnetic hypersensitivity (EHS) is a poorly understood condition in which patients report symptoms following perceived exposure to weak electromagnetic fields (EMFs) such as those produced by mobile phones or visual display units. Little is known about the aetiology of the condition although experimental data suggest that EMFs are an unlikely causal agent. In this systematic review we assessed the efficacy of any treatment for people reporting EHS. METHODS: Twelve literature databases were examined to identify relevant studies. We also hand-searched conference proceedings and examined the reference sections of reviews and other papers. Only clinical trials that compared the efficacy of a potential treatment for EHS against a control condition were included in the review. RESULTS: Nine controlled clinical trials were identified, examining the effects of cognitive behavioural therapy (4 studies), visual display unit screen filters (2 studies), 'shielding' EMF emitters (1 study), supplementary antioxidant therapy (1 study) and acupuncture (1 study). The quality of these studies was limited. Nevertheless, their results suggest that cognitive behavioural therapy is more effective than providing no treatment. None of the other therapies have had their efficacy adequately demonstrated. CONCLUSIONS: The evidence base concerning treatment options for EHS is limited and more research is needed before any definitive clinical recommendations can be made. However, the best evidence currently available suggests that cognitive behavioural therapy is effective for patients who report being hypersensitive to weak EMFs. PMID- 16361871 TI - Tracking scientific interest in the dissociative disorders: a study of scientific publication output 1984-2003. AB - OBJECTIVE: We attempted to track scientific interest in dissociative amnesia and dissociative identity disorder by estimating the annual output of publications regarding these entities over the last 20 years. METHODS: Using a standard medical index, PsycINFO, we counted the number of indexed publications involving dissociative amnesia and dissociative identity disorder listed for each year. We then compared these rates with those of well-established diagnoses such as anorexia nervosa, alcohol abuse, and schizophrenia. We also systematically reviewed all publications involving dissociative amnesia that appeared in 2003. RESULTS: Annual publications involving dissociative amnesia and dissociative identity disorder rose from low levels in the 1980's to a sharp peak in the mid 1990's, followed by an equally sharp decline to only about one quarter of their peak levels by 2002 and 2003. In contrast, all of the 25 comparison diagnoses in our survey showed constant or steadily rising publication rates; none showed the 'bubble' pattern of the dissociative disorders. Of the 34 papers involving dissociative amnesia identified by PsycINFO for 2003, 10 (32%) appeared skeptical of the validity of dissociative amnesia and/or recovered-memory therapy. Despite a detailed search using multiple medical indices and search terms, we could find only 13 explicit cases of individuals with dissociative amnesia worldwide in the 2003 literature. CONCLUSIONS: Dissociative amnesia and dissociative identity disorder have not generated consistent scientific interest over the years, but instead apparently enjoyed a brief period of fashion that now has waned. Overall, our observations suggest that these diagnostic entities presently do not command widespread scientific acceptance. PMID- 16361872 TI - The development of the Toronto Structured Interview for Alexithymia: item selection, factor structure, reliability and concurrent validity. AB - BACKGROUND: Assessments of personality constructs increasingly use self-report and structured interview instruments, which allow for a multimethod measurement approach and decrease specific measurement method bias. The aim of this study was to develop a valid and reliable structured interview for assessing the alexithymia construct. METHODS: Sixty interview questions were written initially, each with a set of scoring criteria and prompts and probes to elicit information assisting in the scoring of the respondents' answers. RESULTS: After pilot testing, the number of questions was reduced to 43, which were administered to 136 community participants and 97 psychiatric outpatients. A series of item and scale analyses further reduced the item pool to 24 items. Principal component analysis and confirmatory factor analysis of these 24 items revealed preliminary evidence of a hierarchical, four-factor structure, with four lower factors nested within two higher-order latent factors. This structural configuration resulted in the Toronto Structured Interview for Alexithymia (TSIA) with two domain scales and four facet scales. The TSIA and its six scales demonstrated acceptable levels of interrater, internal, and retest reliability. The TSIA and its scales correlated modestly but significantly with the 20-item Toronto Alexithymia Scale and its three factor scales, providing some support for the concurrent validity of this interview. CONCLUSION: The TSIA appears to be a promising structured interview for assessing alexithymia. PMID- 16361873 TI - Dissociation as a predictor of cognitive behavior therapy outcome in patients with obsessive-compulsive disorder. AB - BACKGROUND: Previous studies have found a strong association between dissociation and obsessive-compulsive disorder (OCD). The purpose of the present study was to evaluate whether dissociation is a predictor of cognitive behavior therapy (CBT) outcome in patients with OCD. METHODS: Fifty-two patients with OCD were assessed using the Dissociative Experience Scale (DES), the Yale-Brown Obsessive Compulsive Scale and the Beck Depression Inventory. CBT lasted on average 9.5 weeks and included exposure therapy. RESULTS: Patients who dropped out due to noncompliance had higher baseline DES scores and depression scores compared to the 43 patients (83%) who completed the study. Significant OCD symptom reduction at posttreatment was observed in study completers with a large effect size (d = 1.7). More severe OCD symptoms at posttreatment were associated with higher DES scores at baseline, and treatment nonresponders had significantly higher baseline DES scores compared to responders. These associations with outcome were mainly due to the DES subfactor absorption-imaginative involvement. In regression analyses, higher absorption-imaginative involvement scores at baseline predicted poorer CBT outcome, even after controlling for depressive symptoms, comorbid axis I disorders and concomitant psychotropic drugs. CONCLUSIONS: Results from this preliminary study suggest that higher levels of dissociation (particularly absorption-imaginative involvement) in patients with OCD might predict poorer CBT outcome. If our results can be replicated, treatment outcome might be improved by additional interventions for those patients with OCD who indicate high levels of dissociation, for example by using interventions aimed at improving coping with emotionally stressful situations. PMID- 16361874 TI - Bullying girls - changes after brief strategic family therapy: a randomized, prospective, controlled trial with one-year follow-up. AB - BACKGROUND: Many girls bully others. They are conspicuous because of their risk taking behavior, increased anger, problematic interpersonal relationships and poor quality of life. Our aim was to determine the efficacy of brief strategic family therapy (BSFT) for bullying-related behavior, anger reduction, improvement of interpersonal relationships, and improvement of health-related quality of life in girls who bully, and to find out whether their expressive aggression correlates with their distinctive psychological features. METHODS: 40 bullying girls were recruited from the general population: 20 were randomly selected for 3 months of BSFT. Follow-up took place 12 months after the therapy had ended. The results of treatment were examined using the Adolescents' Risk-taking Behavior Scale (ARBS), the State-Trait Anger Expression Inventory (STAXI), the Inventory of Interpersonal Problems (IIP-D), and the SF-36 Health Survey (SF-36). RESULTS: In comparison with the control group (CG) (according to the intent-to-treat principle), bullying behavior in the BSFT group was reduced (BSFT-G from n = 20 to n = 6; CG from n = 20 to n = 18, p = 0.05) and statistically significant changes in all risk-taking behaviors (ARBS), on most STAXI, IIP-D, and SF-36 scales were observed after BSFT. The reduction in expressive aggression (Anger Out scale of the STAXI) correlated with the reduction on several scales of the ARBS, IIP-D, and SF-36. Follow-up a year later showed relatively stable events. CONCLUSIONS: Our findings suggest that bullying girls suffer from psychological and social problems which may be reduced by the use of BSFT. Expressive aggression in girls appears to correlate with several types of risk-taking behavior and interpersonal problems, as well as with health-related quality of life. PMID- 16361875 TI - Influence of anxiety on the course of heart disease after acute myocardial infarction - risk factor or protective function? AB - BACKGROUND: There is a lack of clarity concerning the effect of anxiety on the course of cardiac events following myocardial infarction. Some studies have identified anxiety as a risk factor for further cardiac events. However, it is also considered to be a protective factor, as a higher level of anxiety may improve patient compliance and regular medical checkups. METHODS: A group of 76 patients with myocardial infarction underwent physical examination and had their anxiety levels assessed within the first week, and 31 months after myocardial infarction. Cardiac events were documented during the follow-up period. We investigated the predictive value for the occurrence of cardiac events of sociodemographic, psychological and physical parameters when evaluated within the first week after myocardial infarction. Compliancy and regularity of medical checkups were registered. RESULTS: Cardiac events occurred in 24 patients during a mean follow-up period of 31 months. The group of anxious patients not only suffered more often from cardiac events, these events also occurred earlier than in nonanxious patients. Age, gender, partner status, level of anxiety and comorbid diabetes at the time of first assessment proved to be discriminatory variables between patients suffering further cardiac incidents and those free of further events. Anxious patients were more likely to continue smoking, whereas less anxious patients were more likely to give up smoking. PMID- 16361876 TI - Laughter regulates gene expression in patients with type 2 diabetes. AB - BACKGROUND: Positive emotions influence endocrinological and immunological response. This study examined the effect of laughter,as an expression of positive emotion, in terms of gene expression changes. METHODS: Using a microarray technique, we analyzed the changes in expression of 18,716 genes from peripheral blood leukocytes in patients with type 2 diabetes, which were induced by laughter. RESULTS: Of the 18,716 genes, 23 genes showed significantly different expression changes after listening to the comic story compared to the lecture. Eight were relatively upregulated and 15 were downregulated 1.5 h after the laughing episode. However, these genes did not include genes that are directly involved in blood glucose metabolism. Among the 23 genes discriminated, all 4 genes encoding proteins involved in the immune response and all 4 signal transduction genes were downregulated. Moreover, it is noteworthy that 5 of the 8 relatively upregulated genes were related to the cell cycle, apoptosis, and cell adhesion. CONCLUSIONS: We demonstrated that laughter, which is an expression of positive emotion, is linked to gene expression. However, the finding of this study does not allow reasonable interpretation for the regulation of gene expression by laughter. A more focused study is needed that may identify the candidate genes for the association between physical condition and positive emotion. PMID- 16361877 TI - Clinically based realities. PMID- 16361878 TI - Implant case acceptance: some basic rules. PMID- 16361879 TI - Consensus conference panel report: crown-height space guidelines for implant dentistry-part 1. AB - The International Congress of Oral Implantologists sponsored a consensus conference on the topic of crown height space on June 26 and 27, 2004, in Las Vegas, Nevada. The panel communicated on several occasions before, during, and after the meeting, both as a group and among individuals. A consensus of 1 opinion was not developed for most issues. However, general guidelines emerged related to the topic. The following article is part 1 of a summary of several guidelines that should be of benefit to the profession at large. PMID- 16361880 TI - A new "platinum" standard for bone grafting: autogenous stem cells. AB - Autogenous bone has long been considered the gold standard of all bone grafting materials. However, complications have been associated with autogenous bone harvesting procedures. This article suggests that an alternative approach, grafting with autogenous bone marrow aspirate, may become a new platinum standard. Bone marrow can be extracted from the large flat bones of the body with relative ease and safety, and it provides a rich source of adult stem cells as well as growth factors that facilitate osteogenesis. Mixed with a resorbable matrix or scaffold, bone marrow aspirate has the potential to reconstitute various bony defects in the mouth to reconstruct the severely atrophic maxilla and mandible. The potentiality and plasticity of stem cells have been well documented. PMID- 16361881 TI - A method of sealing perforated sinus membrane and histologic finding of bone substitutes: a case report. AB - To augment the atrophic posterior maxilla, a sinus bone graft has been widely used for sinus floor augmentation. Various bone substitutes have been developed and grafted in the maxillary sinus with and without membranes perforation, although autogenous bone is recommended as a gold standard of grafting materials. Membrane perforation is the most common complication associated with sinus bone graft. To repair a perforation, various methods have been developed. This case report is focused on histologic findings of 1 bovine hydroxyapatite (Bio-Oss; Geistlich Pharma AG, Wolhusen, Switzerland) and 2 kinds of human mineral allograft- Tutoplast cancellous microchips (TutoGen Medical GmbH, Neunkirchen am. Brand Germany), and irradiated allogeniccancellous bone and marrow (ICB; Rocky Mountain Tissue Bank, Aurora, CO) used for sinus graft in the same patient with membrane perforation after various healing periods. Mineral allograft showed favorable new bone regeneration with the repair of membrane perforation. This case report also describes a technique regarding how to repair completely perforated sinus membrane after the removal of a mucocele using human collagen membrane (Tutoplast pericardium; TutoGen Medical GmbH) and fibrin adhesive (Greenplast; Green Cross Co., Youngin, Korea) to stabilize collagen membrane. PMID- 16361882 TI - Arterial supply of maxillary sinus and potential for bleeding complication during lateral approach sinus elevation. AB - There are 3 arteries that supply the maxillary sinus (i.e., the posterior superior alveolar, infraorbital, and posterior lateral nasal arteries), any of which may be encountered during lateral approach sinus elevation surgery. These arteries are ultimate branches of the maxillary artery. Although it has not been reported, there is a theoretical potential for severing an intraosseously located artery during a vigorous curettage for sinus lining elevation in the posterior medial wall of the sinus. Techniques for cessation of bleeding are discussed, such as electrocautery and endoscopic ligation. Head elevation can significantly decrease blood flow to the area. PMID- 16361883 TI - Drill device for sinus lift. AB - A crestal approach for implant placement in deficient alveolar ridges is presented. The set of drills allows the surgeon to lift the membrane at the floor of the maxillary sinus with a bone disc, without the risk of tearing it. The plastic and cutting drills, respectively, are set in just below the floor of the maxillary sinus. The lifting drill is then slowly inserted from the socket, until a desirable length. Implants were hydroxyapatite coated and had a 3.75-mm diameter. This invention suggests that this is a reliable and predictable technique for the prosthetic rehabilitation of the maxillary posterior regions in the presence of anatomic restrictions for implanting. PMID- 16361884 TI - Simultaneous sinus lifting and alveolar distraction of the atrophic maxillary alveolus for implant placement: a preliminary report. AB - This article describes a procedure for performing simultaneous sinus lifting and alveolar distraction to augment an atrophic maxillary alveolus. This technique is a 1-stage operation that is indicated when the amount of native sinus floor bone is minimal (<5 mm). The technique is contraindicated when there is <2 mm of sinus floor,when a 2-stage operation is needed (sinus lifting, alveolar distraction osteogenesis). Postoperative complications are minimal. PMID- 16361885 TI - Mucogingival pouch flap for sandwich bone augmentation: technique and rationale. AB - This article introduces a novel flap design, mucogingival pouch flap (MPF), to enhance the clinical outcome of sandwich bone augmentation. MPF uses a pouch flap reflection via mucogingival junction extension incisions to provide an improved graft retention, minimized membrane exposure, preserved papilla dimension, and soft tissue camouflage for improved esthetics. There are 4 implant-associated buccal dehiscence defects in 3 patients treated with sandwich bone augmentation technique in conjunction with MPF. All cases yielded an adequate new bone thickness of 1.5-3.5 mm as well as a height of 84% to 100% at 6 months. Rationales, indications, contraindications, advantages, and disadvantages for MPF designs are further discussed. PMID- 16361886 TI - The effect of cigarette smoking on dental implants and related surgery. AB - Cigarette smoking is still considered a common habit. Of smokers, increased plaque accumulation, higher incidence of gingivitis and periodontitis, higher rate of tooth loss, and increased resorption of the alveolar ridge have been found in the oral cavity. Cigarette smoking may adversely affect wound healing, and, thus, jeopardize the success of bone grafting and dental implantation. Bone grafts and sinus lift operations are both common and well-documented procedures before dental implant placement. Heat as well as toxic by-products of cigarette smoking, such as nicotine, carbon monoxide, and hydrogen cyanide, have been implicated as risk factors for impaired healing, and, thus, may affect the success and complications of those surgical procedures. An association among dental implants, grafting procedures (i.e., bone grafts, maxillary sinuses augmentation), and history of smoking has been reported. A higher degree of complication, or implant failure rates, were found in smokers with and without bone grafts. The relationship between cigarette smoking and implant-related surgical procedures, including the incidence of complications associated with these procedures, will be described and discussed based on relevant literature and results of our recent studies. PMID- 16361887 TI - Unexpected return of sensation following 4.5 years of paresthesia: case report. AB - Neural damage leads to a transient or persistent alteration, depending on the severity or type of injury sustained. During the last decade, many investigators reported on paresthesia related to dental implants. In this case report, the patient had presented repeatedly with swelling and suppuration, showing typical signs of peri-implantitis. In addition, the implant was placed in proximity to the mental foramen and possibly had traumatized the mental nerve because the patient had had an altered sensation on his left side for the past 4.5 years. After removal of the implant, a significant diminishing of the paresthesia had occurred, described by the patient as a 40% improvement. Further improvement occurred at 6 and 9 months. In this case report, the findings differ from the current literature in that the return of sensation occurred following a prolonged state of paresthesia. This report documents 2 unique findings. First, an area of persistent paresthesia significantly improved 50 months after the initial injury, upon the removal of the offending implant. Second, the placement of another implant in the same vicinity did not result in recurrent paresthesia. PMID- 16361888 TI - Maxillary sinus augmentation using a synthetic cell-binding peptide: a histologic and transmission electron microscopy case study in man. AB - PepGen P-15 is a combination natural anorganic bovine-derived hydroxyapatite matrix coupled with a synthetic cell-binding peptide (P-15). This material has improved bone formation in periodontal osseous defects and bone regenerative procedures. There were 3 specimens retrieved 18 months after a sinus lifting procedure using PepGen P-15. These specimens were treated to be observed under light microscopy and transmission electron microscopy. Light microscopy showed that most of the particles were surrounded by newly formed bone. In some areas, osteoid matrix was present. No acute inflammatory infiltrate was present. In transmission electron microscopy, all phases of bone formation (i.e., osteoid matrix, woven bone, and lamellar bone) were observed in the newly formed bone around the biomaterial particles. In some regions, this newly formed bone seemed to present interdigitations connecting to or entering into the particle surface. To our knowledge, this is the first report presenting data on transmission electron microscopy of PepGen P-15 used in a sinus augmentation procedure in man. Our results confirm previous reports on the clinical effectiveness of this material. PMID- 16361889 TI - Analysis of changes in implant screws subject to occlusal loading: a preliminary analysis. AB - Loosening of the abutment screw can present a problem for the stability of the implant-supported crown or prosthesis. The purpose of this study was to understand the parameters of screw loosening, using an in vitro model, including loss of torque, screw head rotation, changes in screw dimension, and distortion of the implant-abutment joint. Implants (4 x 10 mm) were potted in autopolymerizing blocks. Abutments were placed with screws tightened with a 35 Ncm torque and standardized crowns fabricated. There were 3 implant systems used: Nobel Biocare USA, Inc. (Yorba Linda, CA), 3i Implant Innovations, Inc. (Palm Beach Gardens, FL), and Bio-Lok International, Inc. (Deerfield Beach, FL). Seven samples were tested for each system. Samples were loaded with 300-N loads for 50,000 cycles at 1 Hz. Torque turn audits were performed at 10,000, 25,000, and 50,000 cycles. At the conclusion of the loading, counterclockwise rotation of the abutment screw was measured. The screws were retrieved and measurements made compared with the controls. Finally, 1 sample from each group was embedded in resin, sectioned longitudinally, and examined under the standard error of the mean. The Nobel Biocare system showed a 9.4-Ncm loss of torque from the loading protocol. This result was accompanied by a counterclockwise rotation of 7 degrees and a 200-im elongation of the screw. Finally, there was compression and distortion of the longitudinally sectioned joint architecture observed with the standard error of the mean. From the 3i and Bio-Lok International groups, no loss of torque, counterclockwise rotation, or lengthening of the screws was observed. Intimate adaptation of the joint without distortion was seen in the longitudinal sections. Screw loosening appears to follow specific parameters that include counterclockwise rotation, lengthening of the screw, and distortion of the screw joint. This process is likely associated with both the physical properties of the screw as well as its configuration. PMID- 16361890 TI - Inter-examiner and intra-examiner reliability of the measurement of marginal bone loss around oral implants. AB - The purpose of this study was to test whether a handheld magnifier could be used as an accurate instrument to examine the periapical radiographs and measure the marginal bone levels around dental implants system. A radiologic evaluation of marginal bone loss around 11 Astra Tech (Astra Tech AB, Molndal, Sweden) oral implants was performed. A total of 22-recorded readings of mesial and distal bone loss were performed using a handheld magnifier with a graticule. There were 3 independent examiners who determined radiographic bone assessment. Thevalues of interclass correlation coefficient for the agreement between pairs of examiners showed a relatively high agreement (0.993 for A vs B, 0.995 for B vs C, and 0.995 for A vs C) and the same for intra-examiner reliability result (0.998 for B vs B). These results showed that a handheld magnifier could be used as a simple, reliable, and reproducible method for measuring bone loss around oral implants. PMID- 16361891 TI - Interleukin-2 and interleukin-6 gene promoter polymorphisms, and early failure of dental implants. AB - Single nucleotide polymorphisms in the promoter region of the human interleukin (IL)-2 (T-330G) and IL-6 (G-174C) genes have modified the transcriptional activity of these cytokines and are associated with several diseases. The aim of this study was to investigate the possible relationship between these single nucleotide polymorphisms and early implant failure. A sample of 74 nonsmokers was divided into 2 groups: test group comprising 34 patients (mean age 49.3 years) with >or=1 implants that failed and control group consisting of 40 patients (mean age 43.8 years) with >or=1 healthy implants. Genomic deoxyribonucleic acid from oral mucosa was amplified by polymerase chain reaction and analyzed by restriction fragment length polymorphism. Monte Carlo simulations (P < 0.05) were used to assess differences in allele and genotypes frequencies of the single nucleotide polymorphisms between the 2 groups. No significant differences were observed in the allele and genotypes distribution of both polymorphisms when the 2 groups were compared. The results indicate that polymorphisms in the IL-2 (T 330G) and IL-6 (G-174C) genes are not associated with early implant failure, suggesting that the presence of those single nucleotide polymorphisms does not constitute a genetic risk factor for implant loss in the studied population. PMID- 16361892 TI - Satisfaction with dental implants: a literature review. AB - Recent years have witnessed a consistent trend toward the introduction of patient assessment of different treatment outcomes in dental practice. Patient satisfaction with dental implants was considered among these treatment modalities. Few literature reviews have been published on this topic. This article critically analyzes the concerned topics related to patient satisfaction with dental implant. A MEDLINE search was completed from 1983 to 2004, along with a manual search, to locate related articles on the topic. Dental implants provided promising and predictable results regarding patient satisfaction and various aspects of life assessment. PMID- 16361895 TI - Resuscitation with a hemoglobin-based oxygen carrier after traumatic brain injury. AB - BACKGROUND: Traumatic brain injury (TBI) remains an exclusionary criterion in nearly every clinical trial involving hemoglobin-based oxygen carriers (HBOCs) for traumatic hemorrhage. Furthermore, most HBOCs are vasoactive, and use of pressors in the setting of hemorrhagic shock is generally contraindicated. The purpose of this investigation was to test the hypothesis that low-volume resuscitation with a vasoactive HBOC (hemoglobin glutamer-200 [bovine], HBOC-301; Oxyglobin, BioPure, Inc., Cambridge, MA) would improve outcomes after severe TBI and hemorrhagic shock. METHODS: In Part 1, anesthetized swine received TBI and hemorrhage (30 +/- 2 mL/kg, n = 15). After 30 minutes, lactated Ringer's (LR) solution (n = 5), HBOC (n = 5), or 10 mL/kg of LR + HBOC (n = 5) was titrated to restore systolic blood pressure to > or = 100 mm Hg and heart rate (HR) to < or = 100 beats/min. After 60 minutes, fluid was given to maintain mean arterial pressure (MAP) at > or = 70 mm Hg and heterologous whole blood (red blood cells [RBCs], 10 mL/kg) was transfused for hemoglobin at < or = 5 g/dL. After 90 minutes, mannitol (MAN, 1 g/kg) was given for intracranial pressure > or = 20 mm Hg, LR solution was given to maintain cerebral perfusion pressure at > or = 70 mm Hg, and RBCs were given for hemoglobin of < or = 5 g/dL. In Part 2, after similar TBI and resuscitation with either LR + MAN + RBCs (n = 3) or HBOC alone (n = 3), animals underwent attempted weaning, extubation, and monitoring for 72 hours. RESULTS: In Part 1, relative to resuscitation with LR + MAN + RBCs, LR + HBOC attenuated intracranial pressure (12 +/- 1 mm Hg vs. 33 +/- 6 mm Hg), improved cerebral perfusion pressure in the initial 4 hours (89 +/- 6 mm Hg vs. 60 +/- 3 mm Hg), and improved brain tissue PO2 (34.2 +/- 3.6 mm Hg vs. 16.1 +/- 1.6 mm Hg; all p < 0.05). Cerebrovascular reactivity and intracranial compliance were improved with LR + HBOC (p < 0.05) and fluid requirements were reduced (30 +/- 12 vs. 280 +/- 40 mL/kg; p < 0.05). Lactate and base excess corrected faster with LR + HBOC despite a 40% reduction in cardiac index. With HBOC alone and LR + HBOC, MAP and HR rapidly corrected and remained normal during observation; however, with HBOC alone, lactate clearance was slower and systemic oxygen extraction was transiently increased. In Part 2, resuscitation with HBOC alone allowed all animals to wean and extubate, whereas none in the LR + MAN + RBCs group was able to wean and extubate. At 72 hours, no HBOC animal had detectable neurologic deficits and all had normal hemodynamics. CONCLUSION: The use of HBOC-301 supplemented by a crystalloid bolus was clearly superior to the standard of care (LR + MAN + RBCs) after TBI. This may represent a new indication for HBOCs. Use of HBOC eliminated the need for RBC transfusions and mannitol. The inherent vasopressor effect of HBOCs, especially when used alone, may misguide initial resuscitation, leading to transient poor global tissue perfusion despite restoration of MAP and HR. This suggests that MAP and HR are inadequate endpoints with HBOC resuscitation. HBOC use alone after TBI permitted early extubation and excellent 72-hour outcomes. PMID- 16361896 TI - Comparisons of survival predictions using survival risk ratios based on International Classification of Diseases, Ninth Revision and Abbreviated Injury Scale trauma diagnosis codes. AB - BACKGROUND: We conducted a comparison of methods for predicting survival using survival risk ratios (SRRs), including new comparisons based on International Classification of Diseases, Ninth Revision (ICD-9) versus Abbreviated Injury Scale (AIS) six-digit codes. METHODS: From the Pennsylvania trauma center's registry, all direct trauma admissions were collected through June 22, 1999. Patients with no comorbid medical diagnoses and both ICD-9 and AIS injury codes were used for comparisons based on a single set of data. SRRs for ICD-9 and then for AIS diagnostic codes were each calculated two ways: from the survival rate of patients with each diagnosis and when each diagnosis was an isolated diagnosis. Probabilities of survival for the cohort were calculated using each set of SRRs by the multiplicative ICISS method and, where appropriate, the minimum SRR method. These prediction sets were then internally validated against actual survival by the Hosmer-Lemeshow goodness-of-fit statistic. RESULTS: The 41,364 patients had 1,224 different ICD-9 injury diagnoses in 32,261 combinations and 1,263 corresponding AIS injury diagnoses in 31,755 combinations, ranging from 1 to 27 injuries per patient. All conventional ICD-9-based combinations of SRRs and methods had better Hosmer-Lemeshow goodness-of-fit statistic fits than their AIS based counterparts. The minimum SRR method produced better calibration than the multiplicative methods, presumably because it did not magnify inaccuracies in the SRRs that might occur with multiplication. CONCLUSION: Predictions of survival based on anatomic injury alone can be performed using ICD-9 codes, with no advantage from extra coding of AIS diagnoses. Predictions based on the single worst SRR were closer to actual outcomes than those based on multiplying SRRs. PMID- 16361897 TI - The role of edaravone on the impairment of endothelial barrier function induced by acute oxidative stress in cultured human umbilical vein endothelial cell monolayer. AB - BACKGROUND: The aim was to determine the effects of a novel free radical scavenger, edaravone, 3-methyl-1-phenyl-2-pyrazoline-5-one (ED), against endothelial barrier dysfunction induced by acute oxidative stress in cultured human umbilical vein endothelial cells (HUVECs). METHODS: To estimate the integrity of the HUVEC monolayer, transendothelial electrical resistance (TEER) was measured for 3 hours. We investigated the permeability change of the monolayer by measuring the concentration of fluorescence in isothiocyanate labeled dextran (FITC-Dx), and estimated the degree of oxidative stress in terms of hydrogen peroxide (H2O2) in the apical chambers. RESULTS: The TEER changes in both xanthine oxidase (XO)+xanthine (X) and X/XO+ED group were significantly lower than the control group (p < 0.001). The amount of 4 FITC-Dx in the XO+X group was significantly higher than the control group at 3 hours (p < 0.001). In the XO+X group, the concentration of H2O2 was significantly higher than control and the X/XO+ED group (p < 0.001 each). Thus, edaravone improved the disturbed endothelial barrier function induced by acute oxidative stress. CONCLUSIONS: The permeability increase induced by acute oxidative stress was prevented by free radical scavenger edaravone significantly in vitro. This radical scavenger may have clinical applications to protect against endothelial barrier dysfunction. PMID- 16361898 TI - Phosphodiesterase inhibition decreases nuclear factor-kappaB activation and shifts the cytokine response toward anti-inflammatory activity in acute endotoxemia. AB - BACKGROUND: In sepsis, activation of inflammatory cells and excessive production of proinflammatory cytokines leads to tissue injury, multiple organ failure, and death. We postulated that attenuation but not complete abrogation of hyperinflammation is of clinical benefit in sepsis. Because pentoxifylline (PTX) is known to decrease tumor necrosis factor (TNF)-alpha production and to increase anti-inflammatory cytokine synthesis, we tested the hypothesis that PTX treatment would change the pro- and anti-inflammatory balance and decrease mortality in a murine model of acute endotoxemia. In addition, we investigated the effects of PTX on nuclear factor (NK)-kappaB activation using lipopolysaccharide (LPS) stimulated human peripheral blood mononuclear cells (PBMCs) as a model. METHODS: Sprague-Dawley rats were treated with intravenous saline (sham), LPS (lipopolysaccharide, Escherichia coli serotype 0111:B4, 5 mg/kg), and concomitant injection of LPS + PTX (25 mg/kg). Four- and 24-hour plasma TNF-alpha and interleukin (IL)-10 levels, 4-hour white cell count, and 24-hour mortality rates were assessed. The IL-10/TNF-alpha ratio was also calculated. Human PBMCs were stimulated with LPS (10 microg/mL) and exposed to PTX (20 mM) concomitantly or 15 minutes after LPS stimulation. I-kappaB phosphorylation by Western blot and NF kappaB nuclear translocation by electrophoretic mobility shift assay were assessed. RESULTS: PTX markedly down-regulates TNF-alpha production. IL-10 levels at 4 hours were up-regulated in both LPS and PTX + LPS-treated animals; however, levels were higher in the LPS groups, which paralleled high TNF-alpha levels. In contrast, IL-10 levels at 4 and 24 hours in PTX + LPS-treated animals remained constant, whereas in LPS-treated animals, IL-10 levels at 24 hours were markedly decreased. A shift in the internal milieu balance toward anti-inflammatory activity was confirmed by the calculation of the IL-10/TNF-alpha ratio. These changes were not related to changes in the number of circulating leukocytes. The 24-hour mortality rate was 50% in the LPS group and nil in PTX-treated animals. In LPS-stimulated PBMCs, PTX markedly decreases I-kappaB phosphorylation and NF kappaB nuclear translocation. CONCLUSION: PTX enhances anti-inflammatory activity and decreases mortality in acute endotoxemia. PTX may be an important adjunct to therapies aiming to modulate the inflammatory response in sepsis. PMID- 16361899 TI - Effect of albumin and Hespan on rodent hepatocyte function after hemorrhagic shock and sepsis. AB - BACKGROUND: Sepsis and hemorrhagic shock (HS) cause hypoalbuminemia, with extravascular albumin relocation. Prior studies on cultured rat hepatocytes showed a fall in albumin genesis when exposed to albumin and Hespan (HES) supplementation. This study assesses the effects of increased colloid osmotic pressure on albumin and urea genesis from hepatocytes exposed to HS or septic insult before harvesting. METHODS: Hepatocyte synthesis of albumin was measured in an in vitro collagen sandwich preparation with a seeding density of 1 million cells per dish in Dulbecco's modified Eagle's medium without additional colloid (control cultures). Study groups included control plus 2% bovine serum albumin (BSA), 4% BSA, 2% HES, and 4% HES. RESULTS: The albumin secretion in the control cultures of HS cells rose from 19.03 microg/d/10(6) cells on day 3 to 88.81 microg/d/10(6) cells by day 8 and remained constant. In contrast, albumin genesis in the 2% and 4% BSA groups rose from a significantly higher (p < 0.05) day 3 value of 39.83 microg/d/10(6) cells and 48.76 microg/d/10(6) cells, respectively, to 116.39 microg/d/10(6) cells and 118.76 microg/d/10(6) cells by day 8 and remained fairly constant. Albumin genesis from the HS cells after 2% HES and 4% HES was similar to control HS cells through day 8 but was significantly greater by day 12. Urea genesis from HS cells was greater in the 4% HES group compared with control and the 2% BSA and 4% BSA groups. The albumin genesis in the control cultures of septic cells was 9.32 microg/d/10(6) cells on day 3, rose to 81.46 microg/d/10(6) cells by day 8, and remained fairly constant. The BSA-supplemented septic cells had significantly higher albumin output than control cells early in the experiment, whereas the 2% HES group had higher albumin secretion than control cells throughout the study. CONCLUSION: HS and septic cultured hepatocytes respond to a rise in interstitial colloid osmotic pressure with a rise in albumin genesis, associated with morphologic changes typical of cellular injury. The mechanism remains cryptic; studies on human hepatocytes are needed. PMID- 16361900 TI - Hextend attenuates hypercoagulability after severe liver injury in swine. AB - BACKGROUND: Hypercoagulability is a major source of morbidity and mortality after injury. A resuscitation regimen that modulates this coagulopathy may prove beneficial. We sought to evaluate the effects of lactated Ringer's (LR) solution and Hextend on the resuscitation of uncontrolled hemorrhagic shock. METHODS: Twenty swine underwent invasive line placement, midline celiotomy, and splenectomy. After a 15-minute stabilization period, we recorded a baseline mean arterial pressure and created a grade V liver injury. The animals bled freely for 30 minutes, after which we measured the initial blood loss (that after injury). We blindly randomized the swine to receive LR solution or Hextend to achieve and maintain the baseline mean arterial pressure for 90 minutes postinjury. Laboratory values were obtained at baseline and on completion of the 2-hour study period. RESULTS: The initial blood loss (before resuscitation) was 22 mL/kg in both treatment groups (p = 0.97). Animals required 119 +/- 78 mL/kg of fluid in the LR group and 40 +/- 21 mL/kg in the Hextend group (p = 0.01). After resuscitation, the secondary blood loss was 3.7 +/- 1.7 mL/kg in the LR group and 4.7 +/- 1.1 mL/kg in the Hextend group (p = 0.1). Thrombelastography revealed a hypercoagulable state in all animals after injury. This was less pronounced in those animals resuscitated with Hextend. Routine tests of coagulation did not reveal a hypercoagulable state. CONCLUSION: Modulation and restoration of normal coagulation is critical in the management of trauma patients. The patient's coagulation profile might determine the type of fluid to be used at various times during their course. Thrombelastography is superior to routine coagulation assays for the detection of a hypercoagulable state. Resuscitation with Hextend results in a decreased fluid requirement and attenuation of hypercoagulability after injury without increased blood loss. PMID- 16361901 TI - Traumatic retroperitoneal hematoma spreads through the interfascial planes. AB - BACKGROUND: The purpose of this study was to reevaluate extension of traumatic retroperitoneal hematoma (RH) and related management strategies in light of the new concept of retroperitoneal fascias as interfascial planes communicating with three compartments. METHODS: Diagnostic computed tomographic images of 169 patients with traumatic RH treated between 1997 and 2003 were retrospectively reviewed. The extension of RH was measured in relation to 10 components: 3 compartments and 7 parts of the interfascial planes. On the basis of careful horizontal and vertical assessment of computed tomographic images, distribution, extent, and volume in each component of RH were assessed. RESULTS: In 88.8% of patients, RH was detected in interfascial planes. Interfascial planes absorbed a large amount of hematoma (mean, 223 +/- 309 mL; range, 0-1,519 mL), whereas the anterior and posterior pararenal spaces absorbed only 5 +/- 16 mL and 21 +/- 30 mL, respectively. The volume of RH in interfascial planes accounted for 78.1% of the total volume. In all cases, RH spread within interfascial planes with regularity: transversely by means of retromesenteric planes and vertically by means of combined interfascial planes. Regular extension patterns allowed RH to be classified by bleeding source. RH originating from retrorenal or combined interfascial plane had a poor prognosis; 51.7% of such patients died as a result of uncontrollable hemorrhage. CONCLUSION: RH was based in the interfascial planes, not the three compartments. Our findings that RH extends and is largely confined within interfascial planes, regardless of cause or volume, could be useful in estimating the extent of RH and developing breakthrough strategies for RH. PMID- 16361902 TI - Determinants of futility of administration of recombinant factor VIIa in trauma. AB - BACKGROUND: "Off-label" use of human coagulation factor VIIa (FVIIa) is presently restricted to patients in extremis at our institution. Although bleeding will diminish in most patients, some will still die early as a result of irreversible shock and/or rebleeding. Futile administration of FVIIa significantly increases the economic burden of this expensive therapy and therefore limits its availability. On the basis of both human and in vitro studies, profound acidosis may be expected to predict lack of response. In addition, the depth of hemorrhagic shock, as defined by the degree of hypoperfusion over a given period of time, may be predictive of failure of FVIIa administration. We hypothesized that retrospective review of FVIIa use would identify variables associated with clinical futility. METHODS: Characteristics of patients receiving FVIIa for acute traumatic hemorrhage were identified. Patients were retrospectively stratified into two groups; those who died as a result of acute hemorrhagic shock (nonresponders) and those in whom hemostasis was achieved and sustained (responders). Demographics, laboratory values, transfusion requirements, and outcomes were recorded for all patients. Data were analyzed using the Student's t test to identify the clinical characteristics of nonresponders and stepwise logistic regression was then used to identify independently predictive factors. A classification and regression tree analysis was conducted to develop a decision tree on the basis of our results. RESULTS: Eighty-one patients received FVIIa therapy over a 3-year period. Among the 46 patients treated for acute hemorrhage, there were 26 with blunt and 20 with penetrating mechanisms of trauma. Average age was 35 +/- 15 years, 72% were male, and the average Injury Severity Score was 36 +/- 15. Revised Trauma Score (RTS), lactate, and preadministration prothrombin time (PT) each predicted lack of response (p < 0.05 for each). RTS and PT were independently predictive of failure of response. An RTS of less than 4.09 and a PT of greater than or equal to 17.6 seconds were significantly associated with futile administration of FVIIa. Age was a significant factor in patients with a PT greater than or equal to 17.6 seconds, whereas ISS was significant in patients with an RTS greater than or equal to 4.09. CONCLUSION: Profound acidosis and coagulopathy may predict failure of FVIIa therapy. Depth of hemorrhagic shock, as described by the RTS, was also associated with futile administration. These variables should be considered as potential contraindications to the use of FVIIa. Earlier administration of FVIIa, before the development of massive blood loss and severe shock, may increase the rate of clinical response. PMID- 16361903 TI - An unlucky horseshoe: blunt aortic rupture after horse kick. PMID- 16361904 TI - Use of a single question to screen trauma patients for alcohol dependence. AB - BACKGROUND: Alcohol-dependent trauma patients are known to be at future risk for both recidivism and mortality. Psychological tools exist to screen for alcohol dependent disorders, and among patients with alcohol-dependent disorders, a brief intervention has been shown to modify behavior. However, the training involved and time required to administer these tools may decrease their utility. We explored the possibility that yet simpler screening tools could be used. METHODS: A prospective consecutive study was designed whereby all adult patients admitted to the trauma service were asked to participate in the study. If consent was obtained, one trained member of the research team would apply a standard alcohol misuse screening tool (i.e., the Alcohol Use Disorders Identification Test [AUDIT]). One hundred forty-nine patients met criteria and agreed to participate. Fully completed questionnaires were then subjected to statistical analysis. RESULTS: Among the 149 participating patients, 36% were women and 64% were men, and 146 had blood alcohol levels (BALs) drawn. Those 146 patients form the basis of this report. Fifty-seven of the 146 (39%) patients had positive BALs and, among them, 74% of men and 54% of women recorded screening results consistent with harmful or dependent drinking. Among those with negative blood alcohol levels, 8% of women and 13% of men had scores consistent with harmful or dependent drinking by the AUDIT score. In both groups of patients, a cutoff of three or more drinks per day as a response to question 2 on the AUDIT (i.e., "On a typical day when you are drinking, how many drinks do you have?") correlated strongly with scores on the entire screening tool (AUDIT) in identifying those at risk for alcohol misuse. CONCLUSION: It may be reasonable to substitute a single question for the entire AUDIT screening instrument to screen for those at risk for alcohol misuse. If so, this single question could easily be incorporated into the history taken among patients admitted to a trauma service. Confirmation with a larger study is recommended. PMID- 16361905 TI - Effect of the Uniform Accident and Sickness Policy Provision Law on alcohol screening and intervention in trauma centers. AB - BACKGROUND: Alcohol screening and intervention in trauma centers are widely recommended. The Uniform Accident and Sickness Policy Provision Law (UPPL) exists in most states, and allows insurers to refuse payment for treatment of injuries in patients with a positive alcohol or drug test. This article analyzed the UPPL's impact on screening and reimbursement, measured the knowledge of legislators about substance use problems in trauma centers, and determined their opinions about substance use-related exclusions in insurance contracts for trauma care. METHODS: A nationwide survey of members of the American Association for the Surgery of Trauma was conducted. A separate survey of legislators who are members of the Senate, House, or Assembly and serve in some leadership role on committees responsible for insurance in their state was also performed. RESULTS: Ninety eight trauma surgeon and 56 legislator questionnaires were analyzed. Surgeons' familiarity with the UPPL was limited; only 13% believed they practiced in a UPPL state, but 70% actually did. Despite lack of knowledge of the statute, 24% reported an alcohol- or drug-related insurance denial in the past 6 months. This appeared to affect screening practices; the majority of surgeons (51.5%) do not routinely measure blood alcohol concentration, even though over 91% believe blood alcohol concentration testing is important. Most (82%) indicated that if there were no insurance barriers, they would be willing to establish a brief alcohol intervention program in their center. Legislators were aware of the impact of substance use on trauma centers. They overwhelmingly agreed (89%) that alcohol problems are treatable, and 80% believed it is a good idea to offer counseling in trauma centers. As with surgeons, the majority (53%) were not sure whether the UPPL existed in their state, but they favored prohibiting alcohol-related exclusions by a 2:1 ratio, with strong bipartisan support. CONCLUSIONS: The study documents strong support for screening and intervention programs by both trauma surgeons and legislators. Surgeons experience alcohol-related insurance denials but are not familiar with the state law that sanctions this practice. A majority of legislators are also not familiar with the UPPL but support elimination of insurance statutes that allow exclusion of coverage for trauma care on the basis of intoxication. PMID- 16361906 TI - Scottish urban versus rural trauma outcome study. AB - BACKGROUND: Outcome following trauma and health care access are important components of health care planning. Resources are limited and quality information is required. We set the objective of comparing the outcomes for patients suffering significant trauma in urban and rural environments in Scotland. METHOD: The study was designed as a 2 year prospective observational study set in the west of Scotland, which has a population of 2.58 million persons. Primary outcome measures were defined as the total number of inpatient days, total number of intensive care unit days, and mortality. The participants were patients suffering moderate (ISS 9-15) and major (ISS>15) trauma within the region. The statistical analysis consisted of chi square test for categorical data and Mann Whitney U test for comparison of medians. RESULTS: There were 3,962 urban (85%) and 674 rural patients (15%). Urban patients were older (50 versus 46 years, p = 0.02), were largely male (62% versus 57%, p = 0.02), and suffered more penetrating traumas (9.9% versus 1.9%, p < 0.001). All prehospital times are significantly longer for rural patients (p < 0.001), include more air ambulance transfers (p < 0.001), and are characterized by greater paramedic presence (p < 0.001). Excluding neurosurgical and spinal injuries transfers, there was a higher proportion of transfers in the rural major trauma group (p = 0.002). There were more serious head injuries in the urban group (p = 0.04), and also a higher proportion of urban patients with head injuries transferred to the regional neurosurgical unit (p = 0.037). There were no differences in length of total inpatient stay (median 8 days, p = 0.7), total length of stay in the intensive care unit (median two days, p = 0.4), or mortality (324 deaths, moderate trauma, p = 0.13; major trauma, p = 0.8). CONCLUSION: Long prehospital times in the rural environment were not associated with differences in mortality or length of stay in moderately and severely injured patients in the west of Scotland. This may lend support to a policy of rationalization of trauma services in Scotland. PMID- 16361907 TI - Long-term mortality following trauma: 10 year follow-up in a population-based sample of injured adults. AB - BACKGROUND: The aim of the study was to quantify trauma-related mortality in injured adults over 10 years postinjury. METHODS: A population-based matched cohort study used linked administrative data from Manitoba, Canada, to identify an inception cohort (1988-1991) of hospitalized trauma cases (ICD-9-CM 800-959.9) aged 18-64 years (n = 18,210) and a matched noninjured comparison group (n = 18,210). Mortality outcomes were obtained by linking the two cohorts with the Manitoba Population Registry for a period of 10 years postinjury. RESULTS: The adjusted all-cause mortality rate ratio (MRR) was 7.29 (95% CI 4.53-11.74) for the 60 days immediately postinjury. The MRRs ranged between 1.17 and 2.41 for the remainder of the 10 year follow-up period. The index injury was estimated to be responsible for 41% of all recorded deaths in the injured cohort. CONCLUSIONS: Estimates of the total mortality burden, based on the early inpatient period alone, substantially underestimates the true burden from injury. PMID- 16361908 TI - Effects of reamed or unreamed intramedullary nailing under non-damaged conditions on pulmonary function in sheep. AB - BACKGROUND: Pulmonary damage after reamed or unreamed femoral nailing has been controversial in clinical and experimental studies. We investigated changes in pulmonary function and coagulatory response in normal lungs of sheep after reamed or unreamed intramedullary nailing of an unfractured femur. METHODS: We used three groups of sheep (each group, n = 6): reamed femoral nailing (RFN); unreamed femoral nailing (UFN); and sham control group. Femoral osteotomies and lung contusions were not made. Hemodynamic monitoring data and blood gas data (BGD) were recorded, and blood samples for biochemistry (antithrombin III, lipid peroxidase and fibrinogen) were collected 0 to 6 hours after nailing. Bronchoalveolar lavage fluid (BALF) and myeloperoxidase (MPO) activity in pulmonary tissue were analyzed 6 hours after nailing. RESULTS: There were no significant differences in time course of hemodynamic monitoring data, BGD, or biochemical data among the three groups. In BALF analysis, the lipid-laden cell count in the RFN group was higher than those of the other groups, and MPO activity was highest in the RFN group. CONCLUSION: RFN did not affect pulmonary function at the physiologic level or affect the coagulatory system at the acute stage, but RFN under non-damaged conditions could lead to leukocyte activation in lung tissue without worse alteration of alveolar permeability in the acute post operative course. PMID- 16361909 TI - Emergent stabilization of pelvic ring injuries by controlled circumferential compression: a clinical trial. AB - BACKGROUND: Pelvic ring injuries are associated with a high incidence of mortality mainly due to retroperitoneal hemorrhage. Early stabilization is an integral part of hemorrhage control. Temporary stabilization can be provided by a pelvic sheet, sling, or an inflatable garment. However, these devices lack control of the applied circumferential compression. We evaluated a pelvic circumferential compression device (PCCD), which allows for force-controlled circumferential compression. In a prospective clinical trial, we documented how this device can provide effective reduction of open-book type pelvic injuries without causing overcompression of lateral compression type injuries. METHODS: Sixteen patients with pelvic ring injuries were enrolled. Pelvic fractures were temporarily stabilized with a PCCD until definitive stabilization was provided. Anteroposterior pelvic radiographs were obtained before and after PCCD application, and after definitive stabilization. These radiographs were analyzed to quantify pelvic reduction due to the PCCD in comparison to the quality of reduction after definitive stabilization. Results were stratified into external rotation and internal rotation fracture patterns. RESULTS: In the external rotation group, the PCCD significantly reduced the pelvic width by 9.9 +/- 6.0%. This reduction closely approximated the 10.0 +/- 4.1% reduction in pelvic width achieved by definitive stabilization. In the internal rotation group, the PCCD did not cause significant overcompression. No complications were observed. CONCLUSIONS: A PCCD can effectively reduce pelvic ring injuries. It poses a minimal risk for overcompression and complications as compared with reduction alternatives that do not provide a feedback on the applied reduction force. PMID- 16361910 TI - Modified vastus lateralis flap in treating a difficult hip infection. AB - BACKGROUND: Difficult-to-treat hip infections are typically a reconstructive challenge to orthopedic surgeons. Treatment regimens must be tailored to each patient on the basis of an overall evaluation of systemic host factors and local wound factors. To treat a medically ill patient with compromised local wounds in the hip area, a modified vastus lateralis flap transposition was developed. This flap transposition is of a simple design and effective as both a soft-tissue coverage and a dead space filler. METHODS: From January 2002 to June 2003, 10 patients (5 male patients and 5 female patients) suffering difficult hip infections were referred to Chang Gung Memorial Hospital at Chia-Yi for further treatment. Before visiting the clinic, each patient had undergone repeated failed operations, including debridement, introduction of antibiotic-loaded cement spacers, and fasciocutaneous flap transposition. After being diagnosed with difficult hip infections, these patients were treated with the modified vastus lateralis flap transposition immediately after radical debridement. The average follow-up period was 17.4 months (range, 9-27 months). RESULTS: The hip infections in all 10 patients were controlled and the wounds healed uneventfully. The need for another soft-tissue procedure was not indicated. The C-reactive protein returned to a near-normal and stable level within 1 month. The only complication occurred when case 1 developed a contralateral iliopsoas abscess and secondary septic hip 9 months after the procedure. There was no morbidity or mortality related to the technique. CONCLUSION: For medically ill patients with compromised local wounds, the modified vastus lateralis muscle flap transposition after radical debridement is a simple and effective way of resolving refractory and limb-threatening hip infections. PMID- 16361911 TI - 10-year review of knee dislocations: is arteriography always necessary? AB - BACKGROUND: Historically, arteriography has been used routinely in patients with knee dislocations. Recently, selectivity based on physical examination (PE) has emerged. Critics cite limited clinical evidence. We sought to determine whether PE accurately confirms or excludes surgically significant vascular injuries associated with knee dislocations. METHODS: We conducted an institutional review board-approved retrospective review of patients admitted at a university-based Level I trauma center with knee dislocations from January 1, 1993, to December 31, 2002. Thirty-nine patients (28 male patients and 11 female patients; average age, 42.8 years) had 39 dislocations (27 left and 12 right, 26 posterior and 13 anterior). Most patients (n = 25) were involved in motor vehicle crashes. All patients underwent arteriography. RESULTS: Of the 20 normal arteriograms, all had normal PE. Of the 19 abnormal arteriograms, 8 had a normal PE and 11 were abnormal. Within this subgroup, none of the 8 with normal PE required surgery, whereas 7 of the 11 with abnormal PE required surgery. None of the nonoperative patients had vascular complications during the hospital stay. Sensitivity and specificity were 100% for PE on surgically significant vascular injury. CONCLUSION: Routine arteriography is unnecessary in patients with a normal PE after reduction of the knee dislocation. PMID- 16361912 TI - Diagnosis of occult scaphoid fracture with high-spatial-resolution sonography: a prospective blind study. AB - BACKGROUND: Evaluation of diagnostic accuracy of high-spatial-resolution sonography (HSR-S) in occult scaphoid fractures. PATIENTS AND METHODS: HSR-S was performed in 24 patients with clinically suspected fracture and normal radiographs. Three levels of clinical suspicion were considered (high, intermediate, and low). Three levels of sonographic suspicion were defined on the basis of cortical interruption, radiocarpal effusion, and scapho-trapezium trapezoid effusion. Three positive criteria were interpreted as being highly indicative of fracture. Data from sonograms were compared with computed tomography (CT) scans. RESULTS: CT scanning demonstrated a fracture of the scaphoid in five patients. The global sensitivity of HSR-S for detection of occult scaphoid fracture was 100% and the specificity 79%. All patients with demonstrated occult fracture had a high sonography index of suspicion. A high sonography index of suspicion was correlated with 100% sensitivity, specificity, positive predictive value, and negative predictive value. CONCLUSION: HSR-S is a reliable, available, and cost-effective method in early diagnosis of occult fractures of the scaphoid. The presence of three defined criteria is required to assess the diagnosis. PMID- 16361913 TI - Impaired wound healing after local soft x-ray irradiation in rat skin: time course study of pathology, proliferation, cell cycle, and apoptosis. AB - BACKGROUND: Soft x-ray irradiation is usually used to treat skin-related disease in the clinic, but its effect on wound healing has not been well elucidated. The purpose of our study was to develop a wound-healing model of local soft x-ray irradiation on rats and to clarify the possible cellular mechanisms through which radiation may influence healing. METHODS: Full-thickness, 2-cm-diameter, dorsal cutaneous tissue was excised after local irradiation in rats. The dose-effect curve was recorded, and five animals irradiated with doses of 521 rad were killed at postwound days 3, 6, 9, and 15 and five were healed for histologic examination (hematoxylin and eosin staining), cell proliferation (5-bromo-2'-deoxyuridine [BrdU] incorporation), apoptosis (and terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeling [TUNEL] method), and the cell cycle (flow cytometry). RESULTS: Soft x-ray irradiation delayed wound-healing time with an S like curve, and the wounds that received doses in excess of 700 rad failed to heal within 40 days. This dose-effect range is far below that of gamma and hard x ray irradiation. Relative to control wounds, which contained prominent BrdU positive labeling at days 3 through 9 and minimal TUNEL-positive labeling during the entire healing, the irradiated wounds had less BrdU-positive labeling and significant TUNEL-positive labeling at days 3 through 9 but more BrdU-positive labeling and similar TUNEL-positive labeling at day 15. Flow cytometry studies found a higher proportion of G0/G1 phase cells (days 3-9), a lower (days 3-9) then higher (days 13-22) proportion of S phase cells, and a persistent lower proportion of G2/M phase cells (during the entire healing process) in the irradiated wounds compared with the controls. CONCLUSION: Local soft x-ray irradiation could delay wound healing in a dose-dependent manner and shows a more effective biological effect than that of gamma and hard x-ray irradiation. Radiation-induced inactive cell proliferation, active cell apoptosis, and arrested cell cycle at days 3 through 9 may be one of the cellular mechanism responsible for delayed wound healing. PMID- 16361914 TI - Screening for blunt cerebrovascular injury: evaluating the accuracy of multidetector computed tomographic angiography. AB - BACKGROUND: Digital subtraction angiography (DSA) has been widely considered the "gold standard" for diagnosing blunt cerebral vascular injury, but recent advances in multidetector computed tomographic angiography (CTA) may have led to improved accuracy compared with DSA. METHODS: A retrospective study of patients who underwent CTA and DSA for suspected blunt cerebral vascular injury was performed. Two neuroradiologists and one radiology resident, blinded to the other test, independently reviewed CTA and DSA studies. Sensitivity and specificity of CTA were determined using DSA as the gold standard. RESULTS: Thirty-two patients met the study criteria. Consensus DSA interpretation identified 17 vascular injuries in 15 patients. Among the three radiologists, the sensitivity of CTA for carotid injury ranged from 83% (10 of 12) to 92% (11 of 12), and the specificity ranged from 88% (36 of 41) to 98% (40 of 41). The sensitivity for vertebral artery injury ranged from 40% (2 of 5) to 60% (3 of 5), and the specificity ranged from 90% (35 of 39) to 97% (38 of 39). CTA sensitivity rose to 80% (four of five) in the vertebral artery when a lower diagnostic threshold was used. CONCLUSION: The sensitivity of CTA for carotid artery injury at our center was higher than in previous studies. This might be attributable to the superior quality of multidetector CTA images. Our conclusions on CTA accuracy in the vertebral artery are limited by the small number of injuries. PMID- 16361915 TI - Multiple imputation of the Glasgow Coma Score. AB - BACKGROUND: To investigate whether multiple imputation (MI) of missing Glasgow Coma Scale (GCS) values generates more accurate GCS frequency distributions and less biased parameter estimates in logistic regression models predicting mortality than the standard procedure of excluding observations with missing GCS values. METHODS: The study population consisted of 5,065 patients with complete GCS information from the trauma registry of a Level 1 trauma center. Missing GCS values were imposed on the data set, and the performance of MI (extrapolating missing GCS from a data prediction model) and of deleting all data observations with missing GCS (list-wise deletion) were evaluated. GCS and Trauma and Injury Severity Score (TRISS) frequency distributions and parameter estimates were compared with true values from the original data set. RESULTS: GCS and TRISS frequency values generated by MI were much more accurate than those generated by list-wise deletion. GCS and TRISS parameter estimates generated by MI all had acceptable bias and coverage rates when compared with true values. List-wise deletion provided biased parameter estimates for the GCS, the Revised Trauma Score, and the Injury Severity Score. CONCLUSION: MI is a valid solution to the problem of missing GCS data in trauma research. It allows the conservation of precious data observations and leads to unbiased estimates in consequent analyses. Analyses, which exclude observations with missing GCS data, provide biased results. PMID- 16361916 TI - The effect of ventilator-associated pneumonia on the prognosis of head trauma patients. AB - BACKGROUND: To investigate the effect of ventilator-associated pneumonia (VAP) on the prognosis of head trauma patients. METHODS: We performed a retrospective case control study in which 57 head trauma patients with VAP were matched to 57 head trauma patients without VAP. Matching criteria were age (+/-5 years), Glasgow Coma Scale score (+/-2), Injury Severity Score (+/-5), Simplified Acute Physiology Score II (+/-5), and duration of exposure to mechanical ventilation. RESULTS: The most causative organisms of VAP were Pseudomonas aeruginosa, and Acinetobactor baumannii (36.8% and 33.8% of isolated organisms, respectively). The duration of mechanical ventilation, intensive care unit stay, and hospital stay were significantly increased in case patients (13 +/- 8.4, 24.5 +/- 18, and 30.8 +/- 18.6 days, respectively) compared with control patients (8.3 +/- 4.3, 12.3 +/- 8, and 20.3 +/- 18.7 days, respectively). Mortality rate was also higher in case (29.8%) than in control (12.3%) patients (p = 0.02). CONCLUSION: We conclude that the occurrence of VAP caused by high-risk organisms in cranial trauma patients may increase the risk of death, the mechanical ventilation duration, the intensive care unit stay, and the hospital stay. PMID- 16361917 TI - Large tidal volume mechanical ventilation induces vascular dysfunction in rats. AB - BACKGROUND: Experimental studies have shown that mechanical ventilation using high tidal volumes (V(T)) damages the lungs, causing pulmonary edema. We tested the hypothesis that high V(T) ventilation in rats induces major vascular dysfunction. METHODS: Healthy Sprague-Dawley rats, weighing (mean +/- SD) 340 +/- 15 g, were ventilated with either V(T) = 9 mL/kg and positive end-expiratory pressure (PEEP) = 8 (n = 8) or V(T) = 35 mL/kg and PEEP = 0 (n = 8). The high V(T) used in the injurious ventilation group is in the V(T) range used in other studies to induce lung damage in a short period of time in rats. Lungs were removed for examination under light microscopy and vascular rings from the thoracic aorta were studied for isometric tension recording. RESULTS: Relaxations to acetylcholine (p < 0.001) and sodium nitroprusside (p < 0.05) and contractions to norepinephrine were markedly decreased (p < 0.001) in the high V(T) group, as compared with the low V(T) group. CONCLUSION: Injurious mechanical ventilation in normal rats is associated with vascular dysfunction characterized by decreased relaxation to an endothelium-dependent vasodilator and to a nitrous oxide donor and by decreased response to norepinephrine. PMID- 16361918 TI - The role of acute blood transfusion in the development of acute respiratory distress syndrome in patients with severe trauma. AB - BACKGROUND: Patients with major trauma necessitating the transfusion of packed red blood cells (PRBCs) are at increased risk for the acute respiratory distress syndrome (ARDS). However, it is presently unknown whether the amount of transfused blood is independently associated with development of ARDS in patients with severe trauma. METHODS: This is a prospective cohort study of 102 consecutive patients with severe trauma from an intensive care unit in a Level I trauma center. RESULTS: Patients were divided into three predetermined groups on the basis of the total number of units of PRBCs received in the initial 24 hours. A significant association was identified between an acute exposure to transfused blood and the development of ARDS. Twenty-one percent of patients who received 0 to 5 units of PRBCs developed ARDS, compared with 31% of those patients who received 6 to 10 units of PRBCs and 57% of those who received greater than 10 units of PRBCs (p = 0.007). The association between the amount of transfused blood and the development of ARDS remained significant in a multivariable logistic regression model accounting for differences in severity of illness, type of trauma, race, gender, and base deficit (p = 0.002; odds ratio, 14.4; 95% confidence interval, 3.2-78.7). Patients who received more units of PRBCs during the first 24 hours also had a higher hospital mortality rate (p = 0.03). CONCLUSION: In severely injured trauma patients who require administration of packed red blood cells, the amount of transfused blood is independently associated with both the development of ARDS and hospital mortality. PMID- 16361919 TI - Lawn mower-related injuries to children. AB - BACKGROUND: Despite current prevention efforts, approximately 9,400 children younger than 18 years continue to receive emergency care for lawn mower-related injuries each year in the United States. METHODS: In this study, we analyzed data from a consecutive series of children treated for lawn mower-related injuries during a 53-consecutive-month period in the emergency department of a large, academic children's hospital. The objective of this study was to describe the epidemiology of lawn mower-related injuries to these children and to investigate the beliefs of parents regarding lawn mower use. RESULTS: There were 85 children treated for lawn mower-related injuries during the study period. The mean age was 7.6 years (SD, 4.3 years; median, 5 years), and 65% were boys. Thirty-four patients (40.0%) were admitted to the hospital, including 3 (3.5%) to the pediatric intensive care unit, and 30 (35.3%) required surgical intervention in the operating room. There were 25 children with lacerations (29.4%), 22 with fractures (25.9%), 18 with amputations (21.2%), and 10 with burns (11.8%). The most common body region injured was the lower extremity, accounting for 57.6% (49 of 85) of injuries, including 33 injuries (38.8%) to the foot/toe and 16 injuries (18.8%) to the leg. The hand/finger and head/neck regions each accounted for another 18.8% of injuries. The leading mechanism of injury was run-over/back-over (22.4%), followed by other blade contact (17.6%), thrown object (12.9%), burn (10.6%), and a fall off the mower (7.0%). Lacerations accounted for 68.8% (11 of 16) of injuries to the head/neck compared with 20.3% (14 of 69) to other body regions (p < 0.001; relative risk [RR], 3.39; 95% confidence interval [CI], 1.99 < RR < 6.01). Twelve injuries (36.4% [12 of 33]) to the foot/toe were amputations compared with 6 (11.5% [6 of 52]) to other body regions (p = 0.01; RR, 3.15; 95% CI, 1.31 < RR < 7.58). Burns accounted for 20.9% (9 of 43) of injuries among children 5 years of age or younger compared with 2.4% (1 of 42) of injuries to children older than 5 years of age (p = 0.02; RR, 8.79; 95% CI, 1.16 < RR < 66.39). On average, parents believed that a child should be a minimum of 13 years of age to operate a ride-on mower with supervision. Eighty-six percent of parents indicated that they had made changes in safety practices after the injury event. CONCLUSION: Despite current prevention efforts, serious injuries associated with lawn mowers continue to occur to children. Parental education should promote compliance with the American Academy of Pediatrics policy recommendation that children should be at least 16 years old before operating a ride-on mower. Automatic protection provided by safer product design is the strategy with the highest likelihood of success in preventing these injuries. The voluntary lawn mower safety standard American National Standards Institute/Outdoor Power Equipment Institute B71.1-2003 should be revised to include more rigorous performance provisions regarding prevention of penetration of feet and toes under the mower and into the path of the blades, shielding of hot mower parts from access by young children, and equipping all ride-on lawn mowers with a no-mow-in reverse default feature, with location of the no-mow-in-reverse override switch behind the seating position of the ride-on mower operator. PMID- 16361920 TI - Airbag deployment and improperly restrained children: a lethal combination. AB - BACKGROUND: Airbag deployment is an acknowledged mechanism of serious trauma in children involved in motor vehicle crashes. From a review of national databases, we determined the number and types of fatal and nonfatal injuries to children caused by airbag deployment and child restraint system use. We also reviewed the relevant literature and provide information useful for caregivers and health care professionals in hopes of reducing future injuries. METHODS: We retrospectively reviewed 263 reported cases in which airbag deployment caused fatal or nonfatal injuries in children from reports released by the National Highway and Transportation Safety Administration and the National Pediatric Trauma Registry. Data were collected from January 1993 to December 2002 and imported into a database program for analysis. RESULTS: Of the 263 pediatric injuries caused by airbag deployment, 159 were fatal, and 104 were nonfatal. The peak incidence occurred in 1998, when 58 children were reported injured. Head injuries were most frequent, involving 170 children (64.6%), followed by spinal injuries, involving 100 children (38.0%). For children in their first year of life, head injuries were the sole mechanism of fatality. Of all children studied, only six (2.3%) were properly restrained. CONCLUSION: Airbag deployment in motor vehicle crashes is a well-recognized mechanism of morbidity and mortality in the pediatric population. Most injuries include trauma to the head and spine, which can have significant long-term consequences. Although the reported incidence of such injuries is decreasing, many children are improperly restrained. In our study, only 2.3% of children were properly restrained, suggesting that proper child restraint and seating position could have prevented most injuries. PMID- 16361921 TI - Car-surfing in southwest Ohio: incidence and injuries. AB - BACKGROUND: A local increasing prevalence of injuries due to the unusual mechanism of "car-surfing," during which youths ride on the hood or roof of a vehicle, led us to review our experience with these injuries with the goal of designing an effective prevention program. METHODS: Patient information from the trauma registry at our Level I trauma center was abstracted and analyzed to determine the characteristics of injuries due to this unusual mechanism. RESULTS: Thirty-five patients were identified over a 15-year period. The average age was 19.5 +/- 5.2 years (mean +/- S.D.); 69% were young men. The number of injuries is increasing, with nine injuries identified during the first 5 years, 10 during the next 5 years, and 16 patients during the most recent 5-year period. Fifty-seven percent of the injuries occurred in a rural area and 43% in an urban area. Blood alcohol levels were tested in 29 patients--18 were negative and 11 were positive. Alcohol was more likely to be a factor in older patients with 77.8% (7/9) of those > or = 21 years old having a positive blood alcohol but only 20% (4/20) < or = 20 years old. The most severe injuries were due to head trauma (82.9%), and five patients died. CONCLUSION: This high risk behavior is becoming more prevalent in our region. The information gained from this study will be valuable to our Injury Prevention Center in designing an effective program to reduce such injuries. PMID- 16361922 TI - Traumatic rupture of renal angiomyolipoma managed with angioembolization followed by elective surgery: a report of two cases. PMID- 16361923 TI - The use of recombinant activated factor VII in trauma-associated hemorrhage with crush injury. PMID- 16361924 TI - Superficial femoral vein for arterial reconstruction in trauma. PMID- 16361925 TI - Superficial septic thrombophlebitis. PMID- 16361926 TI - Minimally invasive retrieval of a foreign body after penetrating soft tissue injury. PMID- 16361927 TI - Multimodality monitoring in the management of refractory intracranial hypertension: a case report. PMID- 16361929 TI - Inflammation and the Host Response to Injury, a large-scale collaborative project: Patient-Oriented Research Core--standard operating procedures for clinical care. I. Guidelines for mechanical ventilation of the trauma patient. PMID- 16361930 TI - Digital nerve blocks: a cadaveric study of an unrecognized trauma? PMID- 16361931 TI - A visual, timeline-based display of evidence for emergency thoracotomy. AB - A novel, visual display of selected evidence regarding emergency resuscitative thoracotomy is presented as a brief review. PMID- 16361932 TI - There is more to Ringer than lactate. PMID- 16361933 TI - Isolated jejunal hematoma. PMID- 16361935 TI - Screening index for child abuse. PMID- 16361936 TI - Rat model of femur fracture and cecal ligation and puncture. PMID- 16361937 TI - Concerns by the general surgeons regarding their future within the management of trauma. PMID- 16361938 TI - The goal of trauma and critical care teams includes restitution of the acutely injured patient. PMID- 16361939 TI - Skeletal muscle-derived progenitor cells exhibit neural competence. AB - Skeletal muscle contains heterogenous progenitor cells that give rise to muscle, hematopoietic cells and bone. The exact phenotypic definition of skeletal muscle progenitor cells has not been fully elucidated nor the potential of these cells to differentiate into neurons. Here, we demonstrate that phenotypically homogenous skeletal muscle progenitor cells defined as Lin-CD45-CD117-CD90+ cells express neural stem cell markers and are responsive to neural induction signals. When exposed to neural induction medium containing basic fibroblast growth factor and brain-derived neurotrophic factor, skeletal muscle progenitor cells dramatically changed their cell morphology, became postmitotic and began expressing neuronal markers. These results reveal unexpected potentials of muscle progenitor cells and suggest that these cells may potentially be used in cell based therapies to replace damaged neurons. PMID- 16361940 TI - Handedness-related functional connectivity using low-frequency blood oxygenation level-dependent fluctuations. AB - Functional neuroimaging studies of handedness have traditionally focused on exploring regionally activated discrepancies, yet little is known regarding the effect of handedness on functional connections of brain regions. Here, we reported the relationship between cerebellar functional connectivity patterns and handedness by low-frequency blood oxygenation level-dependent fluctuations. Compared with the right-handers, the left-handers show higher cerebellar prefrontal, cerebellar-parietal, and cerebellar-temporal functional connectivity, and lower cerebellar-limbic connectivity. Previous non-human primate studies have shown that these regions with known contributions to higher cognitive functions have afferent or efferent connections with the cerebellum. Our findings suggest that these observed low-frequency fluctuation correlations may reflect actually anatomic connections between the cerebellum and cerebral cortex, and the cerebellar involvement in higher function may be associated with handedness. PMID- 16361941 TI - Impacts of facial identity and type of emotion on responses of amygdala neurons. AB - The amygdala has been implicated in the processing of emotional expressions. Who makes the emotion and the type of emotion are important in producing appropriate responses. How amygdala neurons are affected by facial identity and type of emotion, however, has not yet been systematically examined. We examined the activity of amygdala neurons using nine monkey stimuli: 3 monkeys x 3 types of emotion. Of the 227 neurons tested, 77 responded to the monkey stimuli. The effects of facial identity and type of emotion on the response magnitude were significant in 48 and 57 neurons, respectively. Both effects were significant in 38 neurons. These results indicate that both facial identity and type of emotion have strong impacts on amygdala functions. PMID- 16361942 TI - Release of tritiated agmatine from spinal synaptosomes. AB - Intrathecal agmatine (decarboxylated arginine) moderates induction of neuropathic pain, spinal cord injury, and opioid tolerance in rodents. An endogenous central nervous system molecule and N-methyl-D-aspartate receptor antagonist/nitric oxide synthase inhibitor, agmatine may be a neuromodulator. We evaluated depolarization induced release of agmatine from purified spinal nerve terminals (synaptosomes). Agmatine immunoreactivity was observed colocalized or closely apposed to some synaptophysin- and/or synaptotagmin-labeled structures. A temperature- and concentration-dependent uptake of [3H]-agmatine into synaptosomes was observed, consistent with an uptake mechanism. Potassium-induced depolarization resulted in release of [3H]-agmatine from the synaptosomes in a Ca2+-dependent manner, consistent with a neuromodulatory function. These results agree with previous reports of agmatine uptake into synaptosomes of the brain and extend those results to include stimulated release and a spinal site of activity. PMID- 16361943 TI - Decreased regional homogeneity in schizophrenia: a resting state functional magnetic resonance imaging study. AB - We used a newly reported regional homogeneity approach to measure the temporal homogeneity of blood oxygen level-dependent signal for exploring the brain activity of schizophrenia in a resting state. The results showed decreased regional homogeneity in schizophrenia, which distributed over the bilateral frontal, temporal, occipital, cerebellar posterior, right parietal and left limbic lobes, similar to the findings reported in previous resting state functional studies. The brain regions that showed decreased regional homogeneity are believed to be involved in the psychopathology and pathophysiology of schizophrenia. Our results indicate that abnormal brain activity of schizophrenia may exist in a resting state and the regional homogeneity may be potentially helpful in understanding the resting state of schizophrenia. PMID- 16361944 TI - White matter integrity of the whole brain is disrupted in first-episode schizophrenia. AB - Diffusion tensor imaging studies in schizophrenia have demonstrated lower diffusion anisotropy within white matter that provides information about brain white matter integrity. We have examined whether white matter is abnormal in first-episode schizophrenia by using diffusion tensor imaging. Twenty-one schizophrenic patients and healthy controls underwent diffusion tensor imaging scans that analyzed by using a rigorous voxel-based approach. We found that fractional anisotropy in white matter of the patients was lower than that in controls at the cerebral peduncle, frontal regions, inferior temporal gyrus, medial parietal lobes, hippocampal gyrus, insula, right anterior cingulum bundle and right corona radiata. These results suggested that white matter integrity of the whole brain was disrupted in early illness onset of schizophrenia. PMID- 16361945 TI - Better time-intensity trade revealed by bilateral giant magnetostrictive bone conduction. AB - Sound lateralization tests were performed to compare the magnet coil bone conduction headphone with the giant magnetostrictive bone-conduction headphone using 18 healthy participants. Although, no significant difference between these bone-conduction headphones was obtained for the interaural time difference and interaural intensity difference, a significant difference was obtained for the time-intensity trade. This revealed that the difference between the headphones is apparent in the integration of the heterogeneous sensations of the time and intensity difference at the cognitive level, but no difference is apparent between the homogeneous sensations of the discrimination of interaural time difference or interaural intensity difference at the sensory level. It was concluded that the difference at the cognitive level indicates the better performance of the giant magnetostrictive headphone. PMID- 16361946 TI - Local mechanical stimulation of the hearing organ by laser irradiation. AB - Light produces force when interacting with matter. Such radiation pressure may be used to accelerate small objects along the beam path of a laser. Here, we demonstrate that a moderately powerful laser can deliver enough force to locally stimulate the hearing organ, in the absence of conventional sound. Damped mechanical oscillations are observed following brief laser pulses, implying that the organ of Corti is locally resonant. This new method will be helpful for probing the mechanical properties of the hearing organ, which have crucial importance for the ear's ability to detect sound. PMID- 16361947 TI - Effects of motor activity on the organization of primary somatosensory cortex. AB - Recent studies have shown that adaptation of representational maps within the primary somatosensory cortex can be induced by task-related motor activity. Here, we explore the relationship between the complexity of the motor task and the extent of task-specific adaptation within the primary somatosensory cortex. We hypothesized that the extent of adaptation increases with the complexity of the motor task. Using neuromagnetic source imaging based on electrical stimulation of the thumb and ring finger, we demonstrate that cortical finger representations are more distant during performance of the pinch finger grip than in a rest condition. Our data suggest that somatosensory cortical maps undergo rapid modulation depending on the task-specific involvement of somatosensory feedback in movements. PMID- 16361948 TI - Maternal care is similar for rat pups subjected to birth hypoxia and for controls. AB - The rat model of global anoxia during cesarean section birth has been used extensively to investigate effects of birth hypoxia on central nervous system function. This study tested whether differential maternal care mediates central nervous system alterations in this model. Maternal care of mixed litters of pups born vaginally, by cesarean section or by cesarean section with anoxia, was assessed. Frequency and duration of licking and grooming by dams were similar for all birth groups. No group differences were observed in order of retrieval, when pups were displaced from the nest. The results indicate that altered central nervous system function in anoxic animals in this model are not due to differential maternal care, but may be mediated by other mechanisms such as direct hypoxic insult to neurons. PMID- 16361949 TI - Astrocytic gene expression profiling upon hypoxia. AB - Astrocytes play a pivotal role in supporting neuronal survival. In order to better understand the contribution of astrocytes towards adaptive mechanisms, gene expression profiles were analyzed after exposure of primary rat astrocyte cultures to normoxic or hypoxic (<3% O2) conditions using high-density oligonucleotide microarrays and quantitative reverse transcriptase polymerase chain reaction. Twenty-five genes were more than 1.5 fold upregulated, whereas 12 genes were more than 1.5-fold downregulated upon hypoxia (P<0.05). Upregulation of established hypoxia-inducible factor 1 target genes as well as novel transcripts related to energy metabolism, astrocyte survival and differentiation, and lipoprotein binding was confirmed by quantitative reverse transcriptase polymerase chain reaction. Further analysis of these genes might provide a better understanding of astrocyte function upon hypoxic conditions. PMID- 16361950 TI - Hypoglycemia influences oligodendrocyte development and myelin formation. AB - Damage to central nervous system white matter is observed following hypoglycemia, raising the possibility that hypoglycemia influences oligodendrocytes and myelination. To examine effects of hypoglycemia on oligodendrocytes and myelin formation, we studied cultured oligodendrocyte precursor cells and cerebellar slice cultures. We observed that with decreasing concentrations of glucose, oligodendrocyte precursor cell proliferation, maturation, and migration decreased. We also observed that hypoglycemia induced apoptotic cell death and activation of caspase-3 in oligodendrocyte precursor cells. Slice culture studies showed that glucose is required for myelinated fiber formation, as with reduction in the glucose concentration, the density of myelinated fibers decreased. Collectively, these data show that hypoglycemia inhibits oligodendrocyte development and myelination and that hypoglycemia triggers apoptotic cell death in oligodendrocyte precursor cells. PMID- 16361951 TI - Association of migraine and ESR1 G325C polymorphism. AB - Migraine is a genetically complex disorder in which sexual hormones influence the phenotype. ESR1 G594A polymorphism has been associated with migraine in Australians. We performed a case-control study with G594A and G325C polymorphisms to determine whether ESR1 is associated with migraine in our population. An association between G594A and migraine could not be demonstrated here. By contrast, we observed that the C325 allele conferred a 1.6 (95% confidence interval=1.1-2.4) higher risk for suffering from migraine in women than the G allele. Women carrying the C352C genotype were over 3 times more likely to suffer from migraine than those carrying the G325G genotype. Therefore, we conclude that ESR1 G325C polymorphism is associated with migraine in our population. PMID- 16361952 TI - Unexpected sensitivity of the human alpha7 neuronal nicotinic acetylcholine receptor to aminoglycosides. AB - The wide use of antibiotics and the development of resistance is a major health concern and, despite their relatively severe side effects, aminoglycoside antibiotics are still used in clinics. Effects of seven aminoglycosides were investigated at the human homomeric alpha7 and heteromeric alpha4beta2 neuronal nicotinic acetylcholine receptors. All aminoglycosides tested inhibited the acetylcholine-evoked responses with more pronounced effects at alpha7 than at alpha4beta2. Neomycin displayed higher blockade with a half inhibition in the nanomolar range at low calcium concentration and in the micromolar range in physiological calcium concentration but still exerted blockade below the concentration used in the clinic. These data suggest that some of their side effects may be attributable to their interactions with neuronal nicotinic acetylcholine receptors. PMID- 16361953 TI - Single and repeated morphine administrations differently regulate expression of N ethylmaleimide-sensitive factor gene in the rat brain. AB - The present study investigated the potential effects of single and repeated morphine treatments on gene expression of N-ethylmaleimide-sensitive protein, a key regulatory protein of neurotransmitter release and synaptic plasticity. After a single morphine injection (10 mg/kg), N-ethylmaleimide-sensitive factor gene expression increased significantly in the cortex, hippocampus and periaqueductal gray. In contrast, N-ethylmaleimide-sensitive factor gene expression was not changed after repeated morphine exposures in these brain regions. Sixteen hours after cessation of repeated morphine administration, N-ethylmaleimide-sensitive factor mRNA level increased again to a level approximate to that of the single morphine injection group in the hippocampus, periaqueductal gray and cerebral cortex. These findings suggest that the N-ethylmaleimide-sensitive factor may be involved in the maladaptive changes in the nervous system after opiate treatment. PMID- 16361954 TI - Amphetamine increases tyrosine kinase-B receptor expression in the dorsal striatum. AB - Neurotrophic signaling is thought to be important for neuroplasticity in certain forebrain regions following psychostimulant exposure. In this study, we found that repeated administration of amphetamine (5 mg/kg, once daily, 5 days) to rats significantly increased tyrosine kinase-B receptor mRNA levels in the striatum, ventral bed nucleus, and piriform cortex. The most robust increase in tyrosine kinase-B expression occurred in dorsal aspects of the striatum, which also showed elevated levels after a single amphetamine injection. These findings indicate that changes in striatal tyrosine kinase-B signaling could play a role in neuroadaptations and behavioral changes induced by amphetamine treatment. PMID- 16361955 TI - Sexually dimorphic changes in the exploratory and habituation profiles of heterozygous neuregulin-1 knockout mice. AB - The neuregulin-1 gene is widely expressed in the central nervous system and is associated with increased risk for schizophrenia. Using an ethologically based approach, the phenotype of neuregulin-1 heterozygous knockout mice was examined by revealing the individual elements of behaviour in the murine repertoire over the prolonged course of interaction with the environment. During initial exploration, neuregulin-1 mutants displayed a phenotype characterized by increases in locomotion and rearing free, with sex-specific alterations in sifting and grooming. Over subsequent habituation, certain initial effects endured while new phenotypic effects emerged, some of which were again sex specific. These studies elaborate a pleiotropic role of neuregulin-1 in development, plasticity and function, including sexual dimorphism, by defining the elemental, temporal and sex-specific characteristics of the neuregulin-1 mutant ethogram. PMID- 16361956 TI - Center of mass attracts attention. AB - Using the spatial cueing technique, this study demonstrates that the center of mass (centroid) of a visual scene has a special ability to attract attention even when there is no object presented at this location. Four boxes formed an imaginary square and were presented to the left or right hemifield. After the cueing in one box, a target appeared in one of the four boxes and, in addition, at centroid. Fastest reaction times were observed at centroid, irrespective of whether this centroid was also occupied by a box. Reaction times at the uncued locations varied according to their relative positions to centroid and fixation. No inhibition of return effect was observed when the cue was at centroid. PMID- 16361957 TI - Preclinical metabolic changes in mouse prion diseases detected by 1H-nuclear magnetic resonance spectroscopy. AB - Magnetic resonance spectroscopy studies in animal models of prion disease are very few and concern terminal stages of infection. In order to study earlier stages of the disease, we used in-vivo magnetic resonance spectroscopy in a mouse model of scrapie and, for the first time, in mice infected with a bovine spongiform encephalopathy strain. In bovine spongiform encephalopathy-infected mice, we observed an increase in myo-inositol preceding clinical signs by 20 days, followed by a decrease in N-acetylaspartate at advanced stages. In scrapie infected mice, changes in N-acetylaspartate and myo-inositol were detected at the beginning of the symptomatic phase. These results show that magnetic resonance spectroscopy is a valuable tool for detecting subtle metabolic changes associated to gliosis and neuronal dysfunction in prion diseases. PMID- 16361958 TI - Effects of catechol-O-methyltransferase Val158Met polymorphism on the cognitive stability and aggression in the first-onset schizophrenic patients. AB - We assessed catechol-O-methyltransferase (COMT) polymorphism in 132 first-onset schizophrenic patients and 80 healthy controls. The relationship between COMT polymorphism and cognitive function, aggression and psychiatric symptoms was tested in the schizophrenic group. COMTL carrier had higher digit span score and lower similarity score than COMTH homozygote. COMTL carrier had higher attention and delusion scores and lower inappropriate affect scores than COMTH homozygote. Attention and delusion scores of COMTL allele were higher than COMTH allele. COMTL group had higher aggression than COMTH homozygote. Our results support the theory that COMTL allele was related with increased tonic dopamine activity and cognitive 'stability', which may induce cognitive inflexibility in schizophrenia. PMID- 16361959 TI - Hormone replacement therapy, brain volumes and white matter in postmenopausal women aged 60-64 years. AB - Research on the structural and functional effects of hormone replacement therapy on the brain has produced inconsistent results. This paper reports on cross sectional associations between hormone replacement therapy use and volumes of brain structures measured using magnetic resonance imaging in 213 postmenopausal women aged 60-64 years recruited from a large population study. Of these, 64 were current hormone replacement therapy users, 69 previous users and 80 had never used hormone replacement therapy. No differences were observed between groups in total grey matter, white matter, hippocampal or amygdalar volumes, severity or volume of white matter hyperintensities, or in different measures of brain atrophy. While acknowledging the limitations of a cross-sectional study, the results argue against hormone replacement therapy being protective against brain changes associated with ageing in women in their early 60s. PMID- 16361961 TI - Natural disaster preparedness: how you can be ready. PMID- 16361960 TI - Linoleic acid derivative DCP-LA improves learning impairment in SAMP8. AB - In the water-maze test, the linoleic acid derivative, 8-[2-(2-pentyl cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA) (1 mg/kg, intraperitoneally), significantly shortened the prolonged latency for accelerated senescence-prone mice 8 (SAMP8), reaching a level similar to the latency for accelerated-senescence-resistant mice 1 (SAMR1) as control. In the open-field test to assess motor activity, it was confirmed that the DCP-LA effect is not due to increased motor activity. In the passive avoidance test to assess fear memory, DCP-LA had no effect on the latency of acquisition and retention for SAMP8. The results of the present study, thus, suggest that DCP-LA could improve age-related learning impairment by enhancing cognitive functions. PMID- 16361963 TI - Getting a good night's sleep: diagnosing and treating insomnia. AB - Sleep takes up one-third of our lives. At least it should. While complaints of insomnia are typically not the precipitating factor for the office visit, patients will reveal their sleeping difficulties when asked a few simple, straightforward questions. Sleep is at the top of the list for leading a physically and mentally healthy lifestyle, and sleep problems are the source of significant co-morbidities. Current research has provided information about the prevalence, morbidity, and diagnosis of insomnia and some tools for behavioral and pharmacological management of this disorder. Nurses with a good understanding of sleep theories can use sleep and rest as an intervention to promote healing after surgery. PMID- 16361965 TI - Treatment of pressure ulcers with topical negative pressure versus traditional wound management methods: a research sampler. AB - The treatment of pressure ulcers is still a matter of debate. The pros and cons of topical negative pressure versus traditional saline wet-to-moist dressings, platelet-derived growth factor, and surgical debridement are discussed in the literature review. This article examines literature that relates to the mechanism of action and efficacy of topical negative pressure. PMID- 16361966 TI - Iatrogenic nerve lesions of the upper extremity. AB - Thirty-three patients are presented who suffer from iatrogenic nerve injury of the upper extremity. Depending on the nature, duration, and degree of nerve injury, we performed neurolysis (16), direct nerve coaptation (4), and nerve grafting (13). Fourteen (70%) of 20 patients presenting with motor nerve injury achieved motor function against resistance. In 26 of 30 patients (87%) with sensory nerve involvement, at least some superficial sensory function was restored. Surgeons should be familiar with clinical and electroneurophysiological tests for early diagnosis of iatrogenic nerve injuries to allow early intervention for maximal nerve recovery. PMID- 16361967 TI - Functional evaluation of the spine in patients with breast hypertrophy. AB - The purpose of the study in this article was to evaluate the physical impairment of patients with breast hypertrophy and to justify the medical necessity of reduction mammaplasty. In 50 women, 20 to 40 years old, with a body mass index below 25, breast size was measured by volumetry and correlated to a functional spine score. This score was determined by the presence of scoliosis, deviated head position, shoulder protraction, shortening of the pectoralis muscle, mobility of the cervical spine, and strength of the abdominal and back extensor muscles. Impairment of the spine in correlation to the breast size was statistically significant (r = 0.71, p < .0001). PMID- 16361969 TI - Thinking outside the box! Enhance patient education by using shared medical appointments. PMID- 16361970 TI - Preparing for breast augmentation: informed consent. PMID- 16361971 TI - Disclosing unanticipated events. PMID- 16361972 TI - Key issues surrounding TAG F314 pressure ulcers guidance to surveyors. PMID- 16361973 TI - Managing the ever-expanding plastic surgery office: part 1. PMID- 16361974 TI - Methicillin-resistant Staphylococcus aureus: no longer just a hospital infection. PMID- 16361980 TI - Genes involved in the neuroendocrine control of normal puberty and abnormal puberty of central origin. AB - There is now compelling evidence that both normal puberty and disturbed pubertal development of central origin are, to a significant extent, determined by genetic factors. Although delayed sexual development can result from a deficient pituitary responsiveness to GnRH caused by mutations in the GnRH receptor gene, until recently the only genetically determined hypothalamic defects known to affect puberty were those caused by mutations in genes required for the migration of gonadotropin releasing hormone (GnRH) neurons, such as KAL1, FGFR1, and NELF. Recently, mutations in a gene termed GPR54 were identified as causing isolated hypogonadotrophic hypogonadism (IHH), due to a functional, instead of a structural hypothalamic defect. Studies in nonhuman primates and rodent models suggest that the functional integrity of the hypothalamic mechanism controlling puberty requires a gene network that includes GPR54. Altogether, these findings indicate that the genetic underpinnings of disturbed pubertal development of central origin are polygenic, rather than specified by a single gene. PMID- 16361981 TI - Testotoxicosis: current viewpoint. AB - Testotoxicosis is a form of gonadotropin-independent (peripheral) precocious puberty in which boys experience early onset and progression of puberty. Patients have accelerated growth, early development of secondary sexual characteristics and usually reduced adult height. Testotoxicosis is caused by an activating mutation of the luteinizing hormone (LH) receptor, leading to increased levels of sex steroids in the context of low LH. Therapy has, therefore, traditionally targeted steroidogenesis. However, the drugs used have been associated with side effects. More recently, a combination of an oral anti-androgen (spironolactone) and an aromatase inhibitor (testolactone) decreased height velocity and improved predicted height. A phase II study in testotoxicosis is currently underway,exploring the combination of a highly selective anti-androgen, bicalutamide, and the potent aromatase inhibitor, anastrozole. These agents are well tolerated in the populations in which they have been studied and effectively inhibit testosterone activity and estrogen production, in adult patients. PMID- 16361982 TI - Bisphosphonate treatment of pediatric bone disease. AB - The science of measuring bone mineral density has developed rapidly and, with it, an improved understanding of the efficacy and safety of various therapeutic interventions in adults. In contrast, the meaning and precision of such measurements in children are equivocal, and the concept of treatment for low bone density in the young patient is still largely undecided. In this report we review the present state of knowledge regarding the use of bisphosphonates during childhood to ameliorate the skeletal abnormalities associated with osteogenesis imperfecta, idiopathic juvenile osteoporosis, fibrous dysplasia of bone and cerebral palsy. Because of the paucity of long-term studies among children regarding the safety and efficacy of these drugs, it is difficult to formulate strong evidence-based recommendations for their use, except perhaps in children with osteogenesis imperfecta. PMID- 16361983 TI - Optic neuropathies in inherited metabolic disorders. AB - Optic neuropathy is a common cause of childhood visual defects and early diagnosis is important for counseling, management and treatment of the underlying conditions. In most cases, careful examination, family history and neuroimaging are sufficient to identify the underlying cause. However, in unexplained cases, in cases where more than one individual in a family is affected and in cases where optic neuropathy is accompanied by other systemic signs and symptoms, a metabolic disorder should be considered. Metabolic disorders generally show a broad range of multisystem clinical symptoms, including eye defects. Here we review the substantial group of metabolic disorders that include optic neuropathies which may aid ophthalmologist, geneticist, neurologist, endocrinologist and other involved specialists in the diagnosis process. PMID- 16361984 TI - Growth and neurodevelopmental outcome of children born to mothers with pregestational and gestational diabetes. AB - Diabetes during pregnancy may be associated with a high rate of congenital anomalies, disturbances of intrauterine growth and often post-natal neurobehavioral abnormalities in the offspring. The latter are associated with pregestational (PGD) as well as with gestational diabetes (GD). In this review we discuss the effects of maternal glucose intolerance on the long-term growth and development of the offspring. In well-controlled diabetes, birth weight is often within normal limits while in partially controlled diabetes newborns are often macrosomic. In PGD mothers with nephropathy, newborns tend to be born prematurely and small for gestational age (SGA). Offspring of diabetic mothers are often large and overweight in comparison to controls. Their long-term development is sometimes impaired. Delayed brain maturity is often observed in newborns of diabetic mothers compared to controls. The IQ scores of the children born to well controlled diabetic mothers are generally similar to that of control children. However, these children perform less well than controls in fine and gross motor functions. They also seem to have a higher rate of inattention and/or hyperactivity as observed by various tests and questionnaires. In our studies we found, in accordance with published literature, a negative correlation between the performance of the children born to mothers with PGD or GD on various neurodevelopmental and behavioral tests and the severity of maternal hyperglycemia as assessed by blood glycosylated hemoglobin levels and acetonuria. IN CONCLUSION: PGD or GD may adversely affect intrauterine and postnatal growth, attention span and motor functions of the offspring, but not their cognitive ability unless complicated by nephropathy or hypertension. These effects are negatively correlated with the degree of maternal glycemic control. PMID- 16361986 TI - Treatment of acute mania. AB - The DSM-IV criteria for mania require: a distinct period that represents a break from pre-morbid functioning, a duration of at least one week, elevated or irritable mood, at least three to four classical manic signs and symptoms and the absence of any physical factors. Although not specifically mentioned in the ICD 10 or the DSM-IV definitions, delusional, hallucinatory, even first-rank, psychotic experiences can occur in mania. Acute mania can be subdivided into classical pure mania, mania with mood-congruent or mood-incongruent psychosis, mixed state and rapid-cycling mania. One quarter to two thirds of all manic episodes are associated with delusions, while 13% to 40% are associated with hallucinations. Mixed episode is a complex syndrome which is difficult to diagnose, has the most prolonged duration of bipolar episodes and more frequent psychotic profile than pure mania with high suicidality and poor response to drugs. Mixed state mania has been well known since Kreapelin and listed in classification systems with criteria that include both a manic and a major depressive episode nearly every day for at least a one-week period. On the other hand, mixed-like episodes that are clearly caused by somatic antidepressant treatment (e.g., medication, electro-convulsive therapy or light therapy) should not contribute toward a diagnosis of Bipolar I Disorder. Although, theoretically, mania is supposed to be resolved within 1-3 months even without treatment, psychiatric hospitalization is very common in especially severe cases due to functional impairment. Current treatments for mania aim to control the agitation, impulsivity, aggression and psychotic symptoms and to help patients regain their pre-morbid functionality. However, the clinical management of mania is challenging as most patients show syndromal remission but incomplete functional recovery after the first episode of mania. PMID- 16361987 TI - Treatment of bipolar depression. AB - Antidepressants have insufficient effect in 20-40% of patients treated for depressive disorders. This is particularly true for psychotic and agitated depression. When administered on a long-term basis, antidepressants cause a switch into mania in 25-40% of patients and induce rapid cycling. Classical antipsychotics have exhibited good therapeutic efficacy in the treatment of various forms of depression, especially psychotic and agitated forms, albeit burdened with many, above all extrapyramidal, side effects. When administered over long periods of time, classical antipsychotics may have a depressogenic effect. Second-generation antipsychotics have started to be increasingly used in this indication for a variety of reasons including: their antidepressant effect attributable to raised concentrations of catecholamines in the prefrontal cortex, their impact on serotonin transmission, their antipsychotic effect due to their mode of action including the mesolimbic blockade of dopamine D2 receptors, and the low incidence of extrapyramidal and other side effects. The following text encompasses the results of controlled trials using second-generation antipsychotics in the treatment of acute depressive disorders. PMID- 16361988 TI - Maintenance treatment of bipolar disorders. AB - Conventional antipsychotic drugs, although efficacious in the treatment of mania, have not demonstrated a significant usefulness in the maintenance treatment of bipolar disorder. This has primarily been due to a tendency to induce depressive symptoms and depressive recurrences in this group of patients in the course of long-term administration. However, the picture has changed following the introduction of second-generation antipsychotics. These drugs have pro-depressant properties (if any) that are much weaker than conventional antipsychotics. Furthermore, their tolerability, especially in long-term treatment, is more favorable compared to classical antipsychotics. Clinical observations of the action profile of second-generation antipsychotic drugs in the treatment of schizophrenia have pointed to a possibility of these agents possessing mood stabilizing properties. The first such suggestion was made by Zarate (1995) in connection with clozapine. The prevention of manic and depressive recurrences in bipolar disorder is a hallmark of the definition of mood-stabilizers. PMID- 16361989 TI - Treatment of rapid cycling bipolar disorders. AB - Rapid cycling (RC) bipolar disorder is defined as four or more affective episodes within one year. RC bipolar disorder constitutes one of the most difficult forms of the illness to treat effectively. According to several studies, RC bipolar patients have more severe symptoms than non-rapid cycling bipolar patients. Most studies indicate that only 10% to 20% of patients with bipolar disorders experience rapid cycling, but this is of great concern to psychiatrists because of its association with treatment refractoriness. Second generation antipsychotics are increasingly being used in the treatment of bipolar disorder. A first step in the management of rapid-cycling bipolar disorder is the thorough assessment of possible medications or environmental factors that may destabilize the disorder and contribute to the recurrence of episodes, increasing cycle frequency, or both. All currently approved antidepressant drugs pose some risk of mood destabilization, but the risk is highest for tricyclic antidepressants. Discontinuation of antidepressants should be the first step in the management of RC patients. PMID- 16362033 TI - Applied systems biology and malaria. AB - One of the goals of systems-biology research is to discover networks and interactions by integrating diverse data sets. So far, systems-biology research has focused on model organisms, which are well characterized and therefore suited to testing new methods. Systems biology has great potential for use in the search for therapies for disease. Here, the potential of systems-biology approaches in the search for new drugs and vaccines to treat malaria is examined. PMID- 16362034 TI - Mae inhibits Pointed-P2 transcriptional activity by blocking its MAPK docking site. AB - During Drosophila melanogaster eye development, signaling through receptor tyrosine kinases (RTKs) leads to activation of a mitogen activated protein tyrosine kinase, called Rolled. Key nuclear targets of Rolled are two antagonistic transcription factors: Yan, a repressor, and Pointed-P2 (Pnt-P2), an activator. A critical regulator of this process, Mae, can interact with both Yan and Pnt-P2 through their SAM domains. Although earlier work showed that Mae derepresses Yan-regulated transcription by depolymerizing the Yan polymer, the mechanism of Pnt-P2 regulation by Mae remained undefined. We find that efficient phosphorylation and consequent activation of Pnt-P2 requires a three-dimensional docking surface on its SAM domain for the MAP kinase, Rolled. Mae binding to Pnt P2 occludes this docking surface, thereby acting to downregulate Pnt-P2 activity. Docking site blocking provides a new mechanism whereby the cell can precisely modulate kinase signaling at specific targets, providing another layer of regulation beyond the more global changes effected by alterations in the activity of the kinase itself. PMID- 16362035 TI - The two Drosophila cytochrome C proteins can function in both respiration and caspase activation. AB - Cytochrome C has two apparently separable cellular functions: respiration and caspase activation during apoptosis. While a role of the mitochondria and cytochrome C in the assembly of the apoptosome and caspase activation has been established for mammalian cells, the existence of a comparable function for cytochrome C in invertebrates remains controversial. Drosophila possesses two cytochrome c genes, cyt-c-d and cyt-c-p. We show that only cyt-c-d is required for caspase activation in an apoptosis-like process during spermatid differentiation, whereas cyt-c-p is required for respiration in the soma. However, both cytochrome C proteins can function interchangeably in respiration and caspase activation, and the difference in their genetic requirements can be attributed to differential expression in the soma and testes. Furthermore, orthologues of the apoptosome components, Ark (Apaf-1) and Dronc (caspase-9), are also required for the proper removal of bulk cytoplasm during spermatogenesis. Finally, several mutants that block caspase activation during spermatogenesis were isolated in a genetic screen, including mutants with defects in spermatid mitochondrial organization. These observations establish a role for the mitochondria in caspase activation during spermatogenesis. PMID- 16362036 TI - The GRIP1:IRF3 interaction as a target for glucocorticoid receptor-mediated immunosuppression. AB - Glucocorticoids dramatically inhibit cytokine and chemokine production. They act through the glucocorticoid receptor (GR), a ligand-dependent transcription factor that binds to and represses activities of other DNA-bound regulators, activator protein 1 and nuclear factor kappaB, utilizing a p160 GRIP1 as a corepressor. A yeast two-hybrid screen with the GRIP1 corepression domain (RD) yielded interferon (IFN) regulatory factor (IRF)3-a downstream effector of Toll-like receptors (TLR) 3/4 and an essential activator of several IFN and chemokine genes. We defined the GRIP1:IRF3 interface and showed that endogenous GRIP1 and IRF3 interact in mammalian cells. Interestingly, GR and IRF3 competed for GRIP1 binding; GR activation or GRIP1 knockdown in macrophages blocked whereas GRIP1 overexpression rescued IRF3-dependent gene expression. GR interference persisted in MyD88- and IFNA receptor-deficient mice, suggesting a specific disruption of TLR3-IRF3 pathway, not of autocrine IFN signaling. Finally, IRF3-stimulated response elements were necessary and sufficient for TLR3-dependent induction and glucocorticoid inhibition. Thus, GRIP1 plays a cofactor role in innate immunity. Competition with GR for GRIP1 antagonizes IRF3-mediated transcription, identifying the GRIP1:IRF3 interaction as a novel target for glucocorticoid immunosuppression. PMID- 16362037 TI - Molecular basis of RNA recognition by the human alternative splicing factor Fox 1. AB - The Fox-1 protein regulates alternative splicing of tissue-specific exons by binding to GCAUG elements. Here, we report the solution structure of the Fox-1 RNA binding domain (RBD) in complex with UGCAUGU. The last three nucleotides, UGU, are recognized in a canonical way by the four-stranded beta-sheet of the RBD. In contrast, the first four nucleotides, UGCA, are bound by two loops of the protein in an unprecedented manner. Nucleotides U1, G2, and C3 are wrapped around a single phenylalanine, while G2 and A4 form a base-pair. This novel RNA binding site is independent from the beta-sheet binding interface. Surface plasmon resonance analyses were used to quantify the energetic contributions of electrostatic and hydrogen bond interactions to complex formation and support our structural findings. These results demonstrate the unusual molecular mechanism of sequence-specific RNA recognition by Fox-1, which is exceptional in its high affinity for a defined but short sequence element. PMID- 16362038 TI - Novel roles of Akt and mTOR in suppressing TGF-beta/ALK5-mediated Smad3 activation. AB - Insulin-like growth factor-I inhibits transforming growth factor-beta (TGF-beta) signaling by blocking activation of Smad3 (S3), via a phosphatidylinositol 3 kinase (PI3K)/Akt-dependent pathway. Here we provide the first report that the kinase activity of Akt is necessary for its ability to suppress many TGF-beta responses, including S3 activation and induction of apoptosis. Wild-type and myristoylated Akts (Akt(WT) and Akt(Myr)) suppress TGF-beta-induced phospho activation of S3 but not Smad2 (S2), whereas kinase-dead Akt1 (Akt1K179M) or dominant-negative PI3K enhances TGF-beta-induced phospho-activation of both S2 and S3. Using siRNA, rapamycin (Rap), and adenoviral expression for FKBP12 resistant and constitutively active TGF-beta type I receptor (ALK5), we demonstrate that mammalian target of Rap (mTOR) mediates Akt1 suppression of phospho-activation of S3. These and further data on Akt1-S3 binding do not support a recently proposed model that Akt blocks S3 activation through physical interaction and sequestration of S3 from TGF-beta receptors. We propose a novel model whereby Akt suppresses activation of S3 in an Akt kinase-dependent manner through mTOR, a likely route for loss of tumor suppression by TGF-beta in cancers. PMID- 16362039 TI - CLIP-170 facilitates the formation of kinetochore-microtubule attachments. AB - CLIP-170 is a microtubule 'plus end tracking' protein involved in several microtubule-dependent processes in interphase. At the onset of mitosis, CLIP-170 localizes to kinetochores, but at metaphase, it is no longer detectable at kinetochores. Although RNA interference (RNAi) experiments have suggested an essential role for CLIP-170 during mitosis, the molecular function of CLIP-170 in mitosis has not yet been revealed. Here, we used a combination of high-resolution microscopy and RNAi-mediated depletion to study the function of CLIP-170 in mitosis. We found that CLIP-170 dynamically localizes to the outer most part of unattached kinetochores and to the ends of growing microtubules. In addition, we provide evidence that a pool of CLIP-170 is transported along kinetochore microtubules by the dynein/dynactin complex. Interference with CLIP-170 expression results in defective chromosome congression and diminished kinetochore microtubule attachments, but does not detectibly affect microtubule dynamics or kinetochore-microtubule stability. Taken together, our results indicate that CLIP 170 facilitates the formation of kinetochore-microtubule attachments, possibly through direct capture of microtubules at the kinetochore. PMID- 16362040 TI - A subcomplex of RNA polymerase III subunits involved in transcription termination and reinitiation. AB - While initiation of transcription by RNA polymerase III (Pol III) has been thoroughly investigated, molecular mechanisms driving transcription termination remain poorly understood. Here we describe how the characterization of the in vitro transcriptional properties of a Pol III variant (Pol IIIdelta), lacking the C11, C37, and C53 subunits, revealed crucial information about the mechanisms of Pol III termination and reinitiation. The specific requirement for the C37-C53 complex in terminator recognition was determined. This complex was demonstrated to slow down elongation by the enzyme, adding to the evidence implicating the elongation rate as a critical determinant of correct terminator recognition. In addition, the presence of the C37-C53 complex required the simultaneous addition of C11 to Pol IIIdelta for the enzyme to reinitiate after the first round of transcription, thus uncovering a role for polymerase subunits in the facilitated recycling process. Interestingly, we demonstrated that the role of C11 in recycling was independent of its role in RNA cleavage. The data presented allowed us to propose a model of Pol III termination and its links to reinitiation. PMID- 16362041 TI - Identification of MEKK2/3 serine phosphorylation site targeted by the Toll-like receptor and stress pathways. AB - Members of the mitogen-activated protein kinase kinase kinase (MAP3K) family are crucial for the Toll-like receptor (TLR) signaling and cellular stress responses. However, the molecular mechanisms underlying the TLR- and cellular stress mediated MAP3K activation remain largely unknown. In this study, we identified a key regulatory phosphorylation site, serine 519 and serine 526, in MAP3K MEKK2 and MEKK3, respectively. Mutation of this serine to an alanine severely impaired MEKK2/3 activation. We generated an anti-p-MEKK2/3 antibody and used this antibody to demonstrate that lipopolysaccharide induced MEKK2 and MEKK3 phosphorylation on their regulatory serine. We found that the serine phosphorylation was crucial for TLR-induced interleukin 6 production and this process is regulated by TRAF6, a key adaptor molecule for the TLR pathway. We further demonstrated that many, but not all, MAPK agonists induced the regulatory serine phosphorylation, suggesting an involvement of different MAP3Ks in activation of the MAPK cascades leading to different cellular responses. In conclusion, this study reveals a novel molecular mechanism for MEKK2/3 activation by the TLR and cellular stress pathways. PMID- 16362042 TI - Structural basis for Gas6-Axl signalling. AB - Receptor tyrosine kinases of the Axl family are activated by the vitamin K dependent protein Gas6. Axl signalling plays important roles in cancer, spermatogenesis, immunity, and platelet function. The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. Structure based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. PMID- 16362043 TI - Structure of the two most C-terminal RNA recognition motifs of PTB using segmental isotope labeling. AB - The polypyrimidine tract binding protein (PTB) is a 58 kDa protein involved in many aspects of RNA metabolism. In this study, we focused our attention on the structure of the two C-terminal RNA recognition motifs (RRM3 and RRM4) of PTB. In a previous study, it was found that the two RRMs are independent in the free state. We recently determined the structure of the same fragment in complex with RNA and found that the two RRMs interact extensively. This difference made us re evaluate in detail the free protein structure and in particular the interdomain interface. We used a combination of NMR spectroscopy and segmental isotopic labeling to unambiguously study and characterize the interdomain interactions. An improved segmental isotopic labeling protocol was used, enabling us to unambiguously identify 130 interdomain NOEs between the two RRMs and to calculate a very precise structure. The structure reveals a large interdomain interface, resulting in a very unusual positioning of the two RRM domains relative to one another. PMID- 16362044 TI - Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. AB - Bacterial speck disease in tomato is caused by Pseudomonas syringae pv. tomato. Resistance to this disease is conferred by the host Pto kinase, which recognizes P. s. pv. tomato strains that express the effector AvrPto. We report here that an AvrPto-dependent Pto-interacting protein 3 (Adi3) is a member of the AGC family of protein kinases. In mammals, AGC kinases are regulated by 3-phosphoinositide dependent protein kinase-1 (Pdk1). We characterized tomato Pdk1 and showed that Pdk1 and Pto phosphorylate Adi3. Gene silencing of Adi3 in tomato causes MAPKKKalpha-dependent formation of necrotic lesions. Use of a chemical inhibitor of Pdk1, OSU-03012, also implicates Pdk1 and Adi3 in plant cell death regulation. Adi3 thus appears to function analogously to the mammalian AGC kinase protein kinase B/Akt by negatively regulating cell death via Pdk1 phosphorylation. We speculate that the negative regulatory function of Adi3 might be subverted by interaction with Pto/AvrPto, leading to host cell death that is associated with pathogen attack. PMID- 16362045 TI - Huckebein-mediated autoregulation of Glide/Gcm triggers glia specification. AB - Cell specification in the nervous system requires patterning genes dictating spatio-temporal coordinates as well as fate determinants. In the case of neurons, which are controlled by the family of proneural transcription factors, binding specificity and patterned expression trigger both differentiation and specification. In contrast, a single gene, glide cell deficient/glial cell missing (glide/gcm), is sufficient for all fly lateral glial differentiation. How can different types of cells develop in the presence of a single fate determinant, that is, how do differentiation and specification pathways integrate and produce distinct glial populations is not known. By following an identified lineage, we here show that glia specification is triggered by high glide/gcm expression levels, mediated by cell-specific protein-protein interactions. Huckebein (Hkb), a lineage-specific factor, provides a molecular link between glide/gcm and positional cues. Importantly, Hkb does not activate transcription; rather, it physically interacts with Glide/Gcm thereby triggering its autoregulation. These data emphasize the importance of fate determinant cell specific quantitative regulation in the establishment of cell diversity. PMID- 16362046 TI - The fidelity of translation initiation: reciprocal activities of eIF1, IF3 and YciH. AB - Eukaryotic initiation factor eIF1 and the functional C-terminal domain of prokaryotic initiation factor IF3 maintain the fidelity of initiation codon selection in eukaryotes and prokaryotes, respectively, and bind to the same regions of small ribosomal subunits, between the platform and initiator tRNA. Here we report that these nonhomologous factors can bind to the same regions of heterologous subunits and perform their functions in heterologous systems in a reciprocal manner, discriminating against the formation of initiation complexes containing codon-anticodon mismatches. We also show that like IF3, eIF1 can influence initiator tRNA selection, which occurs at the stage of ribosomal subunit joining after eIF5-induced hydrolysis of eIF2-bound GTP. The mechanisms of initiation codon and initiator tRNA selection in prokaryotes and eukaryotes are therefore unexpectedly conserved and likely involve related conformational changes induced in the small ribosomal subunit by factor binding. YciH, a prokaryotic eIF1 homologue, could perform some of IF3's functions, which justifies the possibility that YciH and eIF1 might have a common evolutionary origin as initiation factors, and that IF3 functionally replaced YciH in prokaryotes. PMID- 16362047 TI - XBP-1 is required for biogenesis of cellular secretory machinery of exocrine glands. AB - The secretory function of cells relies on the capacity of the endoplasmic reticulum (ER) to fold and modify nascent polypeptides and to synthesize phospholipids for the subsequent trafficking of secretory proteins through the ER Golgi network. We have previously demonstrated that the transcription factor XBP 1 activates the expression of certain ER chaperone genes and initiates ER biogenesis. Here, we have rescued the embryonic lethality of XBP-1 deficient fetuses by targeting an XBP-1 transgene selectively to hepatocytes (XBP-1-/ ;LivXBP1). XBP-1-/-;LivXBP1 mice displayed abnormalities exclusively in secretory organs such as exocrine pancreas and salivary gland that led to early postnatal lethality from impaired production of pancreatic digestive enzymes. The ER was poorly developed in pancreatic and salivary gland acinar cells, accompanied by decreased expression of ER chaperone genes. Marked apoptosis of pancreatic acinar cells was observed during embryogenesis. Thus, the absence of XBP-1 results in an imbalance between the cargo load on the ER and its capacity to handle it, leading to the activation of ER stress-mediated proapoptotic pathways. These data lead us to propose that XBP-1 is both necessary and sufficient for the full biogenesis of the secretory machinery in exocrine cells. PMID- 16362048 TI - Crystal structure of a clip-domain serine protease and functional roles of the clip domains. AB - Clip-domain serine proteases (SPs) are the essential components of extracellular signaling cascades in various biological processes, especially in embryonic development and the innate immune responses of invertebrates. They consist of a chymotrypsin-like SP domain and one or two clip domains at the N-terminus. Prophenoloxidase-activating factor (PPAF)-II, which belongs to the noncatalytic clip-domain SP family, is indispensable for the generation of the active phenoloxidase leading to melanization, a major defense mechanism of insects. Here, the crystal structure of PPAF-II reveals that the clip domain adopts a novel fold containing a central cleft, which is distinct from the structures of defensins with a similar arrangement of cysteine residues. Ensuing studies demonstrated that PPAF-II forms a homo-oligomer upon cleavage by the upstream protease and that the clip domain of PPAF-II functions as a module for binding phenoloxidase through the central cleft, while the clip domain of a catalytically active easter-type SP plays an essential role in the rapid activation of its protease domain. PMID- 16362049 TI - A 'Collagen Hug' model for Staphylococcus aureus CNA binding to collagen. AB - The structural basis for the association of eukaryotic and prokaryotic protein receptors and their triple-helical collagen ligand remains poorly understood. Here, we present the crystal structures of a high affinity subsegment of the Staphylococcus aureus collagen-binding CNA as an apo-protein and in complex with a synthetic collagen-like triple helical peptide. The apo-protein structure is composed of two subdomains (N1 and N2), each adopting a variant IgG-fold, and a long linker that connects N1 and N2. The structure is stabilized by hydrophobic inter-domain interactions and by the N2 C-terminal extension that complements a beta-sheet on N1. In the ligand complex, the collagen-like peptide penetrates through a spherical hole formed by the two subdomains and the N1-N2 linker. Based on these two structures we propose a dynamic, multistep binding model, called the 'Collagen Hug' that is uniquely designed to allow multidomain collagen binding proteins to bind their extended rope-like ligand. PMID- 16362051 TI - PCNA functions as a molecular platform to trigger Cdt1 destruction and prevent re replication. AB - Ubiquitin-mediated proteolysis of the replication licensing factor Cdt1 (Cdc10 dependent transcript 1) in S phase is a key mechanism that limits DNA replication to a single round per cell cycle in metazoans. In Xenopus egg extracts, Cdt1 is destroyed on chromatin during DNA replication. Here, we report that replication dependent proteolysis of Cdt1 requires its interaction with proliferating cell nuclear antigen (PCNA), a homotrimeric processivity factor for DNA polymerases. Cdt1 binds to PCNA through a consensus PCNA-interaction motif that is conserved in Cdt1 of all metazoans, and removal of PCNA from egg extracts inhibits replication-dependent Cdt1 destruction. Mutation of the PCNA-interaction motif yields a stabilized Cdt1 protein that induces re-replication. DDB1, a component of the Cul4 E3 ubiquitin ligase that mediates human Cdt1 proteolysis in response to DNA damage, is also required for replication-dependent Cdt1 destruction. Cdt1 and DDB1 interact in extracts, and DDB1 chromatin loading is dependent on the binding of Cdt1 to PCNA, which indicates that PCNA docking activates the pre formed Cdt1-Cul4(DDB1) ligase complex. Thus, PCNA functions as a platform for Cdt1 destruction, ensuring efficient and temporally restricted inactivation of a key cell-cycle regulator. PMID- 16362050 TI - Interplay between 7SK snRNA and oppositely charged regions in HEXIM1 direct the inhibition of P-TEFb. AB - Transcription elongation of eukaryotic genes by RNA polymerase II depends on the positive transcription elongation factor b (P-TEFb). When sequestered into the large complex, P-TEFb kinase activity is inhibited by the coordinate actions of 7SK small nuclear RNA (7SK snRNA) and hexamethylene bisacetamide (HMBA)-induced protein 1 (HEXIM1). We found that the basic region in HEXIM1 directs its nuclear import via two monopartite and two bipartite nuclear localization sequences. Moreover, the arginine-rich motif within it is essential for its binding to 7SK snRNA, P-TEFb, and inhibition of transcription. Notably, the basic region interacts with the adjacent acidic regions in the absence of RNA. The removal of the positive or negative charges from these regions in HEXIM1 leads to its sequestration into the large complex and inhibition of transcription independently of the arginine-rich motif. Finally, the removal of the negative charges from HEXIM1 results in its subnuclear localization into nuclear speckles. We propose a model where the interplay between 7SK snRNA and oppositely charged regions in HEXIM1 direct its binding to P-TEFb and subcellular localization that culminates in the inhibition of transcription. PMID- 16362052 TI - Dishevelled mediates ephrinB1 signalling in the eye field through the planar cell polarity pathway. AB - An important step in retinal development is the positioning of progenitors within the eye field where they receive the local environmental signals that will direct their ultimate fate. Recent evidence indicates that ephrinB1 functions in retinal progenitor movement, but the signalling pathway is unclear. We present evidence that ephrinB1 signals through its intracellular domain to control retinal progenitor movement into the eye field by interacting with Xenopus Dishevelled (Xdsh), and by using the planar cell polarity (PCP) pathway. Blocking Xdsh translation prevents retinal progeny from entering the eye field, similarly to the morpholino-mediated loss of ephrinB1 (ref. 2). Overexpression of Xdsh can rescue the phenotype induced by loss of ephrinB1, and this rescue (as well as a physical association between Xdsh and ephrinB1) is completely dependent on the DEP (Dishevelled, Egl-10, Pleckstrin) domain of Xdsh. Similar gain- and loss-of function experiments suggest that Xdsh associates with ephrinB1 and mediates ephrinB1 signalling through downstream members of the PCP pathway during eye field formation. PMID- 16362053 TI - In situ trapping of activated initiator caspases reveals a role for caspase-2 in heat shock-induced apoptosis. AB - Activation of 'initiator' (or 'apical') caspases-2, -8 or -9 (refs 1-3) is crucial for induction of apoptosis. These caspases function to activate executioner caspapses that, in turn, orchestrate apoptotic cell death. Here, we show that a cell-permeable, biotinylated pan-caspase inhibitor (bVAD-fmk) both inhibited and 'trapped' the apical caspase activated when apoptosis was triggered. As expected, only caspase-8 was trapped in response to ligation of death receptors, whereas only caspase-9 was trapped in response to a variety of other apoptosis-inducing agents. Caspase-2 was exclusively activated in heat shock-induced apoptosis. This activation of caspase-2 was also observed in cells protected from heat-shock-induced apoptosis by Bcl-2 or Bcl-xL. Reduced sensitivity to heat-shock-induced death was observed in caspase-2(-/-) cells. Furthermore, cells lacking the adapter molecule RAIDD failed to activate caspase 2 after heat shock treatment and showed resistance to apoptosis in this setting. This approach unambiguously identifies the apical caspase activated in response to apoptotic stimuli, and establishes caspase-2 as a proximal mediator of heat shock-induced apoptosis. PMID- 16362056 TI - Optofluidic control using photothermal nanoparticles. AB - Photothermal metallic nanoparticles have attracted significant attention owing to their energy-conversion properties. Here, we introduce an optofluidic application based on a direct optical-to-hydrodynamic energy conversion using suspended photothermal nanoparticles near the liquid-air interface. Using light beams with submilliwatt power, we can drive and guide liquid flow in microfluidic channels to transport biomolecules and living cells at controlled speeds and directions. Previously, a variety of methods for controlling microscale liquid flow have been developed owing to the increasing interest for microfluidics-based biochemical analysis systems. However, our method dispenses with the need for complex pump and valve devices, surface chemistry and electrode patterning, or any other further effort towards substrate fabrication. Instead, our optofluidic control method will allow the fabrication of all-optical large-scale integrated microfluidic circuits for biomolecular and cellular processing without any physical valve or mechanical pumping device. PMID- 16362057 TI - Histone demethylation by a family of JmjC domain-containing proteins. AB - Covalent modification of histones has an important role in regulating chromatin dynamics and transcription. Whereas most covalent histone modifications are reversible, until recently it was unknown whether methyl groups could be actively removed from histones. Using a biochemical assay coupled with chromatography, we have purified a novel JmjC domain-containing protein, JHDM1 (JmjC domain containing histone demethylase 1), that specifically demethylates histone H3 at lysine 36 (H3-K36). In the presence of Fe(ii) and alpha-ketoglutarate, JHDM1 demethylates H3-methyl-K36 and generates formaldehyde and succinate. Overexpression of JHDM1 reduced the level of dimethyl-H3-K36 (H3K36me2) in vivo. The demethylase activity of the JmjC domain-containing proteins is conserved, as a JHDM1 homologue in Saccharomyces cerevisiae also has H3-K36 demethylase activity. Thus, we identify the JmjC domain as a novel demethylase signature motif and uncover a protein demethylation mechanism that is conserved from yeast to human. PMID- 16362058 TI - Multiplex amplification of the mammoth mitochondrial genome and the evolution of Elephantidae. AB - In studying the genomes of extinct species, two principal limitations are typically the small quantities of endogenous ancient DNA and its degraded condition, even though products of up to 1,600 base pairs (bp) have been amplified in rare cases. Using small overlapping polymerase chain reaction products, longer stretches of sequences or even whole mitochondrial genomes can be reconstructed, but this approach is limited by the number of amplifications that can be performed from rare samples. Thus, even from well-studied Pleistocene species such as mammoths, ground sloths and cave bears, no DNA sequences of more than about 1,000 bp have been reconstructed. Here we report the complete mitochondrial genome sequence of the Pleistocene woolly mammoth Mammuthus primigenius. We used about 200 mg of bone and a new approach that allows the simultaneous retrieval of multiple sequences from small amounts of degraded DNA. Our phylogenetic analyses show that the mammoth was more closely related to the Asian than to the African elephant. However, the divergence of mammoth, African and Asian elephants occurred over a short time, corresponding to only about 7% of the total length of the phylogenetic tree for the three evolutionary lineages. PMID- 16362059 TI - Chromatin organization and cell fate switch respond to positional information in Arabidopsis. AB - Many types of plant cell retain their developmental plasticity and have the capacity to switch fate when exposed to a new source of positional information. In the root epidermis of Arabidopsis, cells differentiate in alternating files of hair cells and non-hair cells, in response to positional information and the activity of the homoeodomain transcription factor GLABRA2 (GL2) in future non hair cells. Here we show by three-dimensional fluorescence in situ hybridization on intact root epidermal tissue that alternative states of chromatin organization around the GL2 locus are required to control position-dependent cell-type specification. When, as a result of an atypical cell division, a cell is displaced from a hair file into a non-hair file, it switches fate. We show that during this event the chromatin state around the GL2 locus is not inherited, but is reorganized in the G1 phase of the cell cycle in response to local positional information. This ability to remodel chromatin organization may provide the basis for the plasticity in plant cell fate changes. PMID- 16362060 TI - NTP technical report on the toxicology and carcinogenesis studies of anthraquinone (CAS No. 84-65-1) in F344/N rats and B6C3F1 mice (Feed Studies). AB - BACKGROUND: Anthraquinone is used to make dyes and paper and as a bird repellant. We studied anthraquinone to determine if it caused cancer in rats or mice. METHODS: We fed groups of 50 male and female rats feed containing 469, 938, 1,875, or 3,750 parts per million (ppm) anthraquinone for 2 years. Similar groups of male and female mice received feed containing 833, 2,500, or 7,500 ppm anthraquinone. Groups of 50 male and female rats and mice receiving undosed feed served as the control groups. Tissues from more than 40 sites were examined for every animal. RESULTS: In each group, the group receiving the highest dose of anthraquinone weighed less than its control group. Male and female rats given anthraquinone had higher rates of tumors of the kidney and urinary bladder. Liver tumors also were increased in female rats and slightly increased in male rats. In male and female mice given anthraquinone, the rates of liver tumors were greatly increased, and a few of these animals developed thyroid gland tumors. CONCLUSIONS: We conclude that anthraquinone caused cancer of the kidney and urinary bladder in male and female rats and of the liver in female rats. The occurrence of some liver tumors in male rats may have been related to anthraquinone exposure. We conclude that anthraquinone caused liver cancer in male and female mice, and thyroid gland tumors in mice may have been related to anthraquinone. PMID- 16362061 TI - Toxicology and carcinogenesis studies of sodium chlorate (Cas No. 7775-09-9) in F344/N rats and B6C3F1 mice (drinking water studies). AB - BACKGROUND: Sodium chlorate occurs when drinking water is disinfected by chlorine dioxide. We studied the effects of sodium chlorate in rats and mice to identify potential toxic or carcinogenic hazards to humans. METHODS: We gave groups of male and female rats drinking water containing 125, 1,000, or 2,000 milligrams (mg) of sodium chlorate per liter (L) of water for two years. Male and female mice received 500, 1,000, or 2,000 mg/L. Other groups of animals received plain tap water and served as the control groups. At the end of the study, tissues from more than 40 sites were examined for every animal. RESULTS: Male and female rats receiving sodium chlorate had higher rates of follicular cell hypertrophy of the thyroid gland, and the groups receiving 2,000 mg/L had higher rates of thyroid gland cancer, compared with the control groups. Female mice exposed to sodium chlorate had a few pancreatic islet cell tumors. CONCLUSIONS: We conclude that sodium chlorate caused some thyroid gland neoplasms in male and female rats. The pancreatic islet cell tumors in female mice may have been related to sodium chlorate exposure. PMID- 16362062 TI - NTP carcinogenesis studies of 2,2-bis(bromomethyl)-1,3-propanediol, nitromethane, and 1,2,3-trichloropropane (cas nos. 3296-90-0, 75-52-5, and 96-18-4) in guppies (Poecilia reticulata) and medaka (Oryzias latipes) (Waterborne Studies). AB - The NTP chose to initiate studies in fish as an exploration of alternate or additional models for examining chemical toxicity and carcinogenicity. The use of small fish species in carcinogenicity testing offered potential advantages as a bioassay test system, including significant savings in cost and time over rodent studies. Large numbers of small fish could be easily maintained in a limited area. The two species chosen for study were guppy (Poecilia reticulata) and medaka (Oryzias latipes), both of which are hardy, easily maintained, and have a low occurrence of background lesions. The three chemicals chosen for study in fish had already been studied by the NTP in rodents, permitting a comparison of results between the two models. Two of the chemicals used (2,2-bis(bromomethyl) 1,3-propanediol and 1,2,3-trichloropropane) were mutagenic and multisite carcinogens in rats and mice. The third chemical, nitromethane, was nonmutagenic with a more modest carcinogenic response in rodents. Male and female guppies and medaka were exposed to 2,2-bis(bromomethyl)- 1,3-propanediol (greater than 99% pure), nitromethane, (greater than 99% pure), or 1,2,3-trichloropropane (99% pure) in aquaria water for up to 16 months. OVERALL STUDY DESIGN: Groups of approximately 220 guppies (two replicates of 110) were maintained in aquaria water containing nominal concentrations of 0, 24, 60, or 150 mg/L 2,2 bis(bromomethyl)-1,3-propanediol; 0, 10, 30, or 70 mg/L nitromethane; or 0, 4.5, 9.0, or 18.0 mg/L 1,2,3-trichloropropane. Groups of approximately 340 medaka (two replicates of 170) were maintained in aquaria water containing 0, 24, 60, or 150 mg/L 2,2-bis(bromomethyl)-1,3-propanediol; 0, 10, 20, or 40 mg/L nitromethane; or 0, 4.5, 9.0, or 18.0 mg/L 1,2,3-trichloropropane. The overall study durations were 16 months for all guppy studies, 14 months for 2,2-bis(bromomethyl)-1,3 propanediol-exposed medaka, and 13 months for nitromethane- and 1,2,3 trichloropropane-exposed medaka. Ten guppies and 10 medaka from each group replicate were sacrificed at 9 months for histopathologic analysis. Approximately one third of the remaining fish from each group were placed in chemical-free water at 9 months and constituted a stop-exposure study component. The remainder of the fish were exposed for the duration of the study and constituted the core study component. A stop-exposure component was added to determine if stopping the exposure at 9 months and transferring to chemical-free aquaria might allow for better survival and tumor development. The sex of guppies and medaka was determined at histopathologic analysis. 2,2-BIS(BROMOMETHYL)-1,3-PROPANEDIOL - 16 MONTH STUDY IN GUPPIES: 2,2-Bis(bromomethyl)-1,3-propanediol was chronically toxic to guppies in the 60 and 150 mg/L core and stop-exposure groups. Due to mortality, exposure of core study animals in the 150 mg/L group was terminated on day 443, after approximately 64 weeks on study, and fish were maintained in 2,2 bis(bromomethyl)- 1,3-propanediol-free water in the exposure system until the end of the study at 69 weeks. Nominal exposure concentrations of 24, 60, and 150 mg/L provided actual aquaria water exposure concentrations of 20.0, 53.5, and 139.0 mg/L 2,2-bis(bromomethyl)- 1,3-propanediol, respectively. There were no treatment related differences between the control and exposed groups in body weights or lengths. At 9 months, hepatocellular adenomas occurred in one 24 mg/L male and in one 150 mg/L male. In the core study, the incidence of hepatocellular adenoma or carcinoma (combined) in 150 mg/L males was greater than that in the controls; multiple adenomas occurred in two 150 mg/L males and in one 150 mg/L female. Cholangioma occurred in a small number of exposed males and females. In the stop exposure study, incidences of hepatocellular adenoma (including multiple) and of hepatocellular carcinoma were greater in 150 mg/L males than in controls. One cholangioma and one cholangiocarcinoma occurred in the 150 mg/L female group. 14 MONTH STUDY IN MEDAKA: Exposure to 2,2-bis(bromomethyl)-1,3-propanediol did not result in any significant reduction in survival, although the mortality of fish was somewhat greater in the 60 and 150 mg/L core study groups than in the control and 24 mg/L groups. After reallocation, mortality of medaka in the 60 and 150 mg/L core groups was slightly increased over the corresponding stop-exposure groups. Nominal exposure concentrations of 24, 60, and 150 mg/L provided actual exposure concentrations of 19.4, 56.9, and 137.8 mg/L 2,2-bis(bromomethyl)- 1,3 propanediol, respectively. Core study animals in the 60 and 150 mg/L groups were significantly larger, in both body length and weight, than control group fish. In the core study, the incidence of hepatocellular adenoma or carcinoma (combined) was increased in 150 mg/L males. Cholangiocarcinomas occurred in a few exposed males and females, with all but one occurring in 150 mg/L fish. One cholangioma occurred in a 150 mg/L female, and one occurred in a control female. In the stop exposure study, incidences of hepatocellular adenoma or carcinoma (combined) were marginally increased in the 150 mg/L group of males and in the 60 and 150 mg/L groups of females as compared with controls. Cholangiocarcinoma occurred in one male and one female in the 150 mg/L groups and in one control female. NITROMETHANE - 16-MONTH STUDY IN GUPPIES: Although the cause of death could not be confirmed in many cases, mortality in the 70 mg/L groups appeared to indicate that this level of nitromethane exposure was chronically toxic. This is confirmed by the similar survival rate of guppies from all treatments following removal from treatment aquaria and placement in stop-exposure. Due to the high mortality of fish in the 70 mg/L core study groups, these fish were removed from treatment (day 396) and fixed for histological analyses after approximately 57 weeks on study. The controls and other exposed groups were sacrificed at 70 weeks. Nominal exposure concentrations of 10, 30, and 70 mg/L provided actual exposure concentrations of 9.9, 28.7, and 66.4 mg/L nitromethane, respectively. There were no treatment-related differences between the control and exposed groups in body lengths or weights. 13-MONTH STUDY IN MEDAKA: Nitromethane in the aquaria supported a substantial microfaunal growth which, without frequent cleaning, affected water quality and treatment concentrations. To maintain acceptable water quality and treatment concentrations potentially affected by the rapid microfaunal growth, the study aquaria were brushed once and siphoned three times each day. Due to this frequent activity, a number of fish probably died due to mechanical injury. Unfortunately, the cause of death could not be confirmed in many cases; the mortality from this activity is believed to have been approximately uniform among treatments and should not have affected the comparison of survival between treatments. Based on mortality in this study and the previous life-span evaluation, the life phase of this study was terminated approximately 13.5 months after hatching. Nominal exposure concentrations of 10, 20, and 40 mg/L resulted in actual exposure concentrations of 9.3, 20.8, and 41.7 mg/L nitromethane, respectively. No differences between control and exposed groups were found in body lengths or weights at the 9-month interim evaluation. Due to mortality, unequal numbers of fish were distributed among the core study and stop-exposure aquaria at 9 months. Differences in lengths and weights were found at 13 months. The biological significance of this finding is unknown. At 9 months, a single cholangiocarcinoma occurred in a 40 mg/L male. Hepatocellular adenomas occurred in two 20 mg/L males and in one 40 mg/L female. In the core study, one cholangioma occurred in a 20 mg/L male, and cholangiocarcinomas were seen in a few exposed males, but none occurred in control males. 1,2,3 TRICHLOROPROPANE - 16-MONTH STUDY IN GUPPIES: The survival of exposed guppies was less than that of the control group at 9 months. Reduced survival was evident at 6 months in the 18.0 mg/L groups and at 7 months in the 4.5 and 9.0 mg/L groups. Survival was significantly reduced in the 18.0 mg/L core study group within 1 month of the 9-month interim evaluation, and mortality in this group was 42.6% between 9 months and study termination. Nominal exposure concentrations of 4.5, 9.0, and 18.0 mg/L resulted in actual exposure concentrations of 4.4, 8.8, and 18.2 mg/L 1,2,3-trichloropropane, respectively. Guppies in the 18.0 mg/L core study group were significantly longer and weighed more than the controls. Fish in the 18.0 mg/L stop-exposure group also weighed more than the controls. Mortality of fish during the study resulted in unequal numbers of individuals distributed to core study and stop-exposure aquaria at 9 months. This appears to have influenced the length and weight of fish measured at study termination (i.e., the smaller tank population allowed the fish to grow more). Observed differences in weight and length between controls and 18.0 mg/L fish was most likely an artifact of the reduced numbers of fish in the 18.0 mg/L aquaria. At 9 months, multiple hepatocellular adenomas occurred in one 4.5 mg/L male, and one hepatocellular adenoma occurred in a control male. In the core study, increased incidences of cholangiocellular (bile duct) and hepatocellular neoplasms occurred in exposed groups of males and females. Cholangioma and cholangiocarcinoma were seen in several exposed males and females. In the stop-exposure study, increased incidences of hepatocellular neoplasms occurred in 18.0 mg/L males and increased incidences of cholangiocellular (bile duct) neoplasms occurred in 18.0 mg/L females. (ABSTRACT TRUNCATED) PMID- 16362063 TI - Year in review: communicable disease surveillance, 2004. PMID- 16362064 TI - Imported malaria notified in New South Wales and the Australian Capital Territory, including trends in notifications of Plasmodium falciparum, 1989 to 2003. PMID- 16362069 TI - Bug Breakfast in the Bulletin. Outbreaks: the past, present and future. PMID- 16362070 TI - Communicable diseases report, March and April 2005. PMID- 16362071 TI - The activity reaction core and plasticity of metabolic networks. AB - Understanding the system-level adaptive changes taking place in an organism in response to variations in the environment is a key issue of contemporary biology. Current modeling approaches, such as constraint-based flux-balance analysis, have proved highly successful in analyzing the capabilities of cellular metabolism, including its capacity to predict deletion phenotypes, the ability to calculate the relative flux values of metabolic reactions, and the capability to identify properties of optimal growth states. Here, we use flux-balance analysis to thoroughly assess the activity of Escherichia coli, Helicobacter pylori, and Saccharomyces cerevisiae metabolism in 30,000 diverse simulated environments. We identify a set of metabolic reactions forming a connected metabolic core that carry non-zero fluxes under all growth conditions, and whose flux variations are highly correlated. Furthermore, we find that the enzymes catalyzing the core reactions display a considerably higher fraction of phenotypic essentiality and evolutionary conservation than those catalyzing noncore reactions. Cellular metabolism is characterized by a large number of species-specific conditionally active reactions organized around an evolutionary conserved, but always active, metabolic core. Finally, we find that most current antibiotics interfering with bacterial metabolism target the core enzymes, indicating that our findings may have important implications for antimicrobial drug-target discovery. PMID- 16362072 TI - Comparing the Dictyostelium and Entamoeba genomes reveals an ancient split in the Conosa lineage. AB - The Amoebozoa are a sister clade to the fungi and the animals, but are poorly sampled for completely sequenced genomes. The social amoeba Dictyostelium discoideum and amitochondriate pathogen Entamoeba histolytica are the first Amoebozoa with genomes completely sequenced. Both organisms are classified under the Conosa subphylum. To identify Amoebozoa-specific genomic elements, we compared these two genomes to each other and to other eukaryotic genomes. An expanded phylogenetic tree built from the complete predicted proteomes of 23 eukaryotes places the two amoebae in the same lineage, although the divergence is estimated to be greater than that between animals and fungi, and probably happened shortly after the Amoebozoa split from the opisthokont lineage. Most of the 1,500 orthologous gene families shared between the two amoebae are also shared with plant, animal, and fungal genomes. We found that only 42 gene families are distinct to the amoeba lineage; among these are a large number of proteins that contain repeats of the FNIP domain, and a putative transcription factor essential for proper cell type differentiation in D. discoideum. These Amoebozoa-specific genes may be useful in the design of novel diagnostics and therapies for amoebal pathologies. PMID- 16362073 TI - Selective constraint on noncoding regions of hominid genomes. AB - An important challenge for human evolutionary biology is to understand the genetic basis of human-chimpanzee differences. One influential idea holds that such differences depend, to a large extent, on adaptive changes in gene expression. An important step in assessing this hypothesis involves gaining a better understanding of selective constraint on noncoding regions of hominid genomes. In noncoding sequence, functional elements are frequently small and can be separated by large nonfunctional regions. For this reason, constraint in hominid genomes is likely to be patchy. Here we use conservation in more distantly related mammals and amniotes as a way of identifying small sequence windows that are likely to be functional. We find that putatively functional noncoding elements defined in this manner are subject to significant selective constraint in hominids. PMID- 16362074 TI - Genome trees from conservation profiles. AB - The concept of the genome tree depends on the potential evolutionary significance in the clustering of species according to similarities in the gene content of their genomes. In this respect, genome trees have often been identified with species trees. With the rapid expansion of genome sequence data it becomes of increasing importance to develop accurate methods for grasping global trends for the phylogenetic signals that mutually link the various genomes. We therefore derive here the methodological concept of genome trees based on protein conservation profiles in multiple species. The basic idea in this derivation is that the multi-component "presence-absence" protein conservation profiles permit tracking of common evolutionary histories of genes across multiple genomes. We show that a significant reduction in informational redundancy is achieved by considering only the subset of distinct conservation profiles. Beyond these basic ideas, we point out various pitfalls and limitations associated with the data handling, paving the way for further improvements. As an illustration for the methods, we analyze a genome tree based on the above principles, along with a series of other trees derived from the same data and based on pair-wise comparisons (ancestral duplication-conservation and shared orthologs). In all trees we observe a sharp discrimination between the three primary domains of life: Bacteria, Archaea, and Eukarya. The new genome tree, based on conservation profiles, displays a significant correspondence with classically recognized taxonomical groupings, along with a series of departures from such conventional clusterings. PMID- 16362075 TI - Invasion by P. falciparum merozoites suggests a hierarchy of molecular interactions. AB - Central to the pathology of malaria disease are the repeated cycles of parasite invasion and destruction of human erythrocytes. In Plasmodium falciparum, the most virulent species causing malaria, erythrocyte invasion involves several specific receptor-ligand interactions that direct the pathway used to invade the host cell, with parasites varying in their dependency on these different pathways. Gene disruption of a key invasion ligand in the 3D7 parasite strain, the P. falciparum reticulocyte binding-like homolog 2b (PfRh2b), resulted in the parasite invading via a novel pathway. Here, we show results that suggest the molecular basis for this novel pathway is not due to a molecular switch but is instead mediated by the redeployment of machinery already present in the parent parasite but masked by the dominant role of PfRh2b. This would suggest that interactions directing invasion are organized hierarchically, where silencing of dominant invasion ligands reveal underlying alternative pathways. This provides wild parasites with the ability to adapt to immune-mediated selection or polymorphism in erythrocyte receptors and has implications for the use of invasion-related molecules in candidate vaccines. PMID- 16362076 TI - Recognition of conserved amino acid motifs of common viruses and its role in autoimmunity. AB - The triggers of autoimmune diseases such as multiple sclerosis (MS) remain elusive. Epidemiological studies suggest that common pathogens can exacerbate and also induce MS, but it has been difficult to pinpoint individual organisms. Here we demonstrate that in vivo clonally expanded CD4+ T cells isolated from the cerebrospinal fluid of a MS patient during disease exacerbation respond to a poly arginine motif of the nonpathogenic and ubiquitous Torque Teno virus. These T cell clones also can be stimulated by arginine-enriched protein domains from other common viruses and recognize multiple autoantigens. Our data suggest that repeated infections with common pathogenic and even nonpathogenic viruses could expand T cells specific for conserved protein domains that are able to cross react with tissue-derived and ubiquitous autoantigens. PMID- 16362077 TI - Ablation of the Sam68 RNA binding protein protects mice from age-related bone loss. AB - The Src substrate associated in mitosis of 68 kDa (Sam68) is a KH-type RNA binding protein that has been shown to regulate several aspects of RNA metabolism; however, its physiologic role has remained elusive. Herein we report the generation of Sam68-null mice by homologous recombination. Aged Sam68-/- mice preserved their bone mass, in sharp contrast with 12-month-old wild-type littermates in which bone mass was decreased up to approximately 75%. In fact, the bone volume of the 12-month-old Sam68-/- mice was virtually indistinguishable from that of 4-month-old wild-type or Sam68-/- mice. Sam68-/- bone marrow stromal cells had a differentiation advantage for the osteogenic pathway. Moreover, the knockdown of Sam68 using short hairpin RNA in the embryonic mesenchymal multipotential progenitor C3H10T1/2 cells resulted in more pronounced expression of the mature osteoblast marker osteocalcin when differentiation was induced with bone morphogenetic protein-2. Cultures of mouse embryo fibroblasts generated from Sam68+/+ and Sam68-/- littermates were induced to differentiate into adipocytes with culture medium containing pioglitazone and the Sam68-/- mouse embryo fibroblasts shown to have impaired adipocyte differentiation. Furthermore, in vivo it was shown that sections of bone from 12-month-old Sam68-/- mice had few marrow adipocytes compared with their age-matched wild-type littermate controls, which exhibited fatty bone marrow. Our findings identify endogenous Sam68 as a positive regulator of adipocyte differentiation and a negative regulator of osteoblast differentiation, which is consistent with Sam68 being a modulator of bone marrow mesenchymal cell differentiation, and hence bone metabolism, in aged mice. PMID- 16362078 TI - The flavoring agent dihydrocoumarin reverses epigenetic silencing and inhibits sirtuin deacetylases. AB - Sirtuins are a family of phylogenetically conserved nicotinamide adenine dinucleotide-dependent deacetylases that have a firmly established role in aging. Using a simple Saccharomyces cerevisiae yeast heterochromatic derepression assay, we tested a number of environmental chemicals to address the possibility that humans are exposed to sirtuin inhibitors. Here we show that dihydrocoumarin (DHC), a compound found in Melilotus officinalis (sweet clover) that is commonly added to food and cosmetics, disrupted heterochromatic silencing and inhibited yeast Sir2p as well as human SIRT1 deacetylase activity. DHC exposure in the human TK6 lymphoblastoid cell line also caused concentration-dependent increases in p53 acetylation and cytotoxicity. Flow cytometric analysis to detect annexin V binding to phosphatidylserine demonstrated that DHC increased apoptosis more than 3-fold over controls. Thus, DHC inhibits both yeast Sir2p and human SIRT1 deacetylases and increases p53 acetylation and apoptosis, a phenotype associated with senescence and aging. These findings demonstrate that humans are potentially exposed to epigenetic toxicants that inhibit sirtuin deacetylases. PMID- 16362079 TI - Genome-wide associations of gene expression variation in humans. AB - The exploration of quantitative variation in human populations has become one of the major priorities for medical genetics. The successful identification of variants that contribute to complex traits is highly dependent on reliable assays and genetic maps. We have performed a genome-wide quantitative trait analysis of 630 genes in 60 unrelated Utah residents with ancestry from Northern and Western Europe using the publicly available phase I data of the International HapMap project. The genes are located in regions of the human genome with elevated functional annotation and disease interest including the ENCODE regions spanning 1% of the genome, Chromosome 21 and Chromosome 20q12-13.2. We apply three different methods of multiple test correction, including Bonferroni, false discovery rate, and permutations. For the 374 expressed genes, we find many regions with statistically significant association of single nucleotide polymorphisms (SNPs) with expression variation in lymphoblastoid cell lines after correcting for multiple tests. Based on our analyses, the signal proximal (cis-) to the genes of interest is more abundant and more stable than distal and trans across statistical methodologies. Our results suggest that regulatory polymorphism is widespread in the human genome and show that the 5-kb (phase I) HapMap has sufficient density to enable linkage disequilibrium mapping in humans. Such studies will significantly enhance our ability to annotate the non-coding part of the genome and interpret functional variation. In addition, we demonstrate that the HapMap cell lines themselves may serve as a useful resource for quantitative measurements at the cellular level. PMID- 16362080 TI - Macrophage migration inhibitory factor: a critical component of autoimmune inflammatory diseases. AB - Autoimmune inflammatory diseases occur commonly in Western populations and include conditions such as juvenile-onset diabetes mellitus, rheumatoid arthritis, inflammatory bowel disease and systemic lupus erythematosus. The precise cause of these diseases remains enigmatic. However, current notions of pathogenesis support an important interplay between host genetics and acquired, or environmental, factors. From an immunologic perspective, autoimmune inflammatory diseases develop as a result of a loss of immune tolerance and the initiation of immune-mediated tissue injury. In the following review, we discuss recent studies pointing to an important role for the upstream mediator macrophage migration inhibitory factor in the effector responses producing autoimmune tissue damage. PMID- 16362081 TI - Leptin and cardiovascular diseases. AB - Although obesity is strongly associated with cardiovascular disease (CVD), the endogenous relationship between obesity and CVD is still not fully clear. Emerging evidence from both animal and human studies indicates that leptin may play an important role in obesity-related CVD. Besides modulating appetite and metabolism, leptin has also been shown to increase sympathetic nerve activity, stimulate generation of reactive oxygen species, upregulate endothelin-1 production and potentiate platelet aggregation. These effects of leptin may contribute to hypertension, endothelial dysfunction and atherosclerosis in obese individuals. Better understanding the mechanisms of leptin resistance should facilitate therapeutic approaches to reverse the phenomenon of selective leptin resistance. These recent discoveries could lead to novel strategies for treatment of obesity-associated CVD. PMID- 16362082 TI - Proline-directed protein kinase FA as a new signal transducing target for lethal cancer treatment. AB - Proline-directed protein kinase F(A) (PDPK F(A)) has been established as a multisubstrate/multifunctional PDPK essential for the development of highly malignant phenotypes and rapid progression of lethal cancers. The recent immunohistochemical, immunocytochemical and clinicopathologic studies combined demonstrate that overexpressed PDPK F(A) is dynamically and closely associated with the most aggressive malignant cells disseminating from primary tumors to peripheral blood, ascites, pleural effusions and second metastatic tumors of various types of cancer patients with poor prognosis. The antisense gene therapeutic studies further demonstrate that overexpressed PDPK F(A) is essential for the development of all aspects of neoplasia including highly metastatic spread, peritoneal dissemination, splenomegaly and chemoradioresistances. The inhibition of cancer progression together with the simultaneous enhancement of chemoradiosensitivities through the suppression of overexpressed PDPK F(A) by specific drug design may work synergistically with surgery and chemoradiotherapy to improve the poor survival rate and life quality of the patients with lethal malignancies. PDPK F(A), therefore, may represent the heel of Achilles and a new promising target for the strategic development of more efficacious treatment for lethal cancers PMID- 16362083 TI - The role of ML-23 and other melatonin analogues in the treatment and management of Parkinson's disease. AB - Contemporary theory regarding the cause and treatment of neuropsychiatric disease strongly suggests that as the human body ages it gradually loses the intrinsic safeguards that protect it from oxidative damage. Melatonin is one hormone that serves this function in that it possesses antioxidative properties in the mammalian body and brain. Melatonin has been shown to prevent the progressive degeneration produced by neurotoxins employed in experimental models to mimic the degenerative events in various neuropsychiatric disease states. There are an abundance of models for numerous disease states demonstrating that melatonin can inhibit oxidative stress and by such a mechanism it is presumed to exert a therapeutic effect. While a similar scenario has been revealed with in vitro work relating specifically to Parkinson's disease, clinical work with melatonin in this disorder demonstrates that it is devoid of any remarkable therapeutic effects. More recent preclinical and clinical work has reliably demonstrated that melatonin in fact may be without therapeutic efficacy and may even worsen the condition. On this pretense, attempts to reduce the bioavailability of melatonin using a melatonin receptor antagonist have been found to completely restore behavioral and regulatory function in the presence of chronically reduced levels of dopamine, without producing side effects commonly seen with traditional dopamine replacement therapy. The unavoidable conclusion from this work suggests that within the dynamic framework of the mammalian brain, hormones may play a duel, and possibly ambivalent, role in homeostasis and in the etiology of disease. Such a position requires a reevaluation of the etiology, the role of dopamine, the neurochemical characteristics of Parkinson's disease and the validity of the models employed to study this and other neuropsychiatric disorders. PMID- 16362084 TI - Chemopreventive functions of isothiocyanates. AB - Numerous animal and epidemiological studies have demonstrated that consumption of cruciferous vegetables is inversely correlated with the cancer incidence. Strong anticarcinogenic activities of cruciferous vegetables are attributed to the high abundance of glucosinolates. Upon consumption, glucosinolates are hydrolyzed into iso-thiocyanates (ITCs), which in turn are conjugated with intracellular glutathione and excreted via mercapturic acid pathway. On the basis of the principle that ITCs can interact with 1,2-benzenedithiole to produce 1,3 benzodithiole-2-thione, a simple high-pressure liquid chromatography-based assay (namely cyclocondensation assay) has been devised to carry out the accurate measurement of the intracellular ITC accumulation. This assay has been adopted to assess the pharmacokinetic profiles of ITCs in humans, to evaluate the enzymatic efficiency of glucosinolate breakdown by myrosinase and to investigate the intracellular absorption and elimination mechanisms of ITCs. In addition, LC-MS MS has recently been introduced as an alternative quantitative method for ITCs and proved to be superior to cyclocondensation assay, in terms of sensitivity as well as selectivity. Among several possible cellular mechanisms, both the modulation of nuclear factor kappaB (NF-kappaB) and activator protein-1 (AP-1) and the induction of phase II cellular detoxification and antioxidant enzymes (in brief phase II enzymes) have been proposed as potential molecular mechanisms of chemoprevention by ITCs. In the present review, we describe how ITCs modulate the activity of NF-kappaB and AP-1 and discuss how Nrf2/ARE module was discovered as the central core in the transcriptional regulation of phase II cellular detoxification and antioxidant enzymes. PMID- 16362085 TI - Molecule of the month. Deferasirox. PMID- 16362086 TI - Recent developments in cardiovascular disease in diabetes mellitus. AB - September 10-15, 2005, the European Association for the Study of Diabetes held its 41st Annual Meeting in Athens, Greece. This meeting was devoted to new developments in understanding, diagnosis and management of diabetes mellitus and related disorders. Each day's program included plenary sessions, symposia, oral presentations, poster discussions, lectures, current issues and supportive care. This article highlights recent developments in pathophysiology, new therapeutic possibilities and preventive aspects of cardiovascular diseases in diabetes mellitus. PMID- 16362087 TI - Vascular remodeling in experimental hypertension. AB - The basic hemodynamic abnormality in hypertension is an increased peripheral resistance that is due mainly to a decreased vascular lumen derived from structural changes in the small arteries wall, named (as a whole) vascular remodeling. The vascular wall is an active, flexible, and integrated organ made up of cellular (endothelial cells, smooth muscle cells, adventitia cells, and fibroblasts) and noncellular (extracellular matrix) components, which in a dynamic way change shape or number, or reorganize in response to physiological and pathological stimuli, maintaining the integrity of the vessel wall in physiological conditions or participating in the vascular changes in cardiovascular diseases such as hypertension. Research focused on new signaling pathways and molecules that can participate in the mechanisms of vascular remodeling has provided evidence showing that vascular structure is not only affected by blood pressure, but also by mechanisms that are independent of the increased pressure. This review will provide an overview of the evidence, explaining some of the pathophysiologic mechanisms participating in the development of the vascular remodeling, in experimental models of hypertension, with special reference to the findings in spontaneously hypertensive rats as a model of essential hypertension, and in fructose-fed rats as a model of secondary hypertension, in the context of the metabolic syndrome. The understanding of the mechanisms producing the vascular alterations will allow the development of novel pharmacological tools for vascular protection in hypertensive disease. PMID- 16362132 TI - Breast cancer risk factors and mammographic breast density in women over age 70. AB - BACKGROUND: Breast density is a strong risk factor for breast cancer, but little is known about factors associated with breast density in women over 70. METHODS: Percent breast density, sex hormone levels and breast cancer risk factor data were obtained on 239 women ages 70-92 recruited from 1986 to 1988 in the United States. Multivariable linear regression was used to develop a model to describe factors associated with percent density. RESULTS: Median (range) percent density among women was 23.7% (0-85%). Body mass index (beta=-0.345, p<0.001 adjusted for age and parity) and parity (beta=-0.277, p<0.001 adjusted for age and BMI) were significantly and inversely associated with percent breast density. After adjusting for parity and BMI, age was not associated with breast density (beta=0.05, p=0.45). Parous women had lower percent density than nulliparous women (23.7 versus 34.7%, p=0.005). Women who had undergone surgical menopause had greater breast density than those who had had a natural menopause (33.4 versus 24.8%, p=0.048), as did women who were not current smokers (26.0 versus 17.3% for smokers, p=0.02). Breast density was not associated with age at menarche, age at menopause, age at first birth, breastfeeding, estrogen levels or androgen levels. In a multivariable model, 24% of the variance in percent breast density was explained by BMI (beta=-0.35), parity (beta=-0.29), surgical menopause (beta=0.13) and current smoking (beta=-0.12). CONCLUSION: Factors associated with breast density in older, post-menopausal women differ from traditional breast cancer risk factors and from factors associated with breast density in pre-menopausal and younger post-menopausal women. PMID- 16362133 TI - [Health economics of psychological diseases]. AB - This article provides an overview of the characteristics and problems of the economic evaluation of mental health care. At first, the problems and methods of measuring direct and indirect costs of mental illness are discussed using cost of illness studies for depression and schizophrenia from the literature as examples. It is shown that, due to methodological problems of measuring costs of the wide range of mental health services as well as productivity losses, various cost of illness studies rendered very different results. Thereafter, the measurement of effects in the economic evaluation of mental health services is discussed whereby the problems of measuring quality-adjusted life years (QALYs) are addressed in particular. Despite the subjectivity of symptoms, preference-based valuation of health-related quality of life has rarely been attempted in psychiatric patients. Finally, frequent flaws of published cost-effectiveness analyses of mental health care are presented. These flaws make the interpretation and comparison of study results difficult. As there are hardly any German cost-effectiveness analyses of mental health care, more research efforts in this field seem to be necessary in Germany. PMID- 16362134 TI - [Health economic aspects of the use of blood and blood products]. AB - An adequate supply of blood components for patients depends on the population's willingness to donate blood as well as on a well-organized blood donation system. In order to ensure the blood supply, a high expenditure is necessary. The article at hand describes two areas of the use of blood and blood products. First, it considers the application of factor concentrates in haemophiliacs (approximately 3000-4000 patients are regularly in need of treatment). For the German statutory health insurance, the estimated yearly expenses for factor concentrates is approx. 450 million Euros, the average costs per patient and year and per treatment center range between 40,903 and 117,597 Euros. The costs for an immune tolerance therapy (ITT) for patients with inhibitors are estimated to be between 70,300 and 3,800,000 Euros for children, depending on the length of the therapy and the degree of severity. In the second part of the article, the authors outline the status quo of the economic evaluation of cellular blood products and present the results of a German cross-sectional survey on the use of erythrocytes in orthopaedic surgery. Within the scope of this hemotherapy study, the transfusion rates vary considerably within the participating treatment centers [elective hip joint replacement (THR): 17 and 94%]. The average blood loss also differs significantly. Comparing the treatment centers, the calculated blood loss during THR shows considerable differences (median blood loss in treatment center with lowest/treatment center with highest blood loss: 883 ml/2,975 ml). The acceptance of low intraoperative hemoglobin values in the clinics is high and diverse. These differences in transfusion procedures between the treatment centers clearly show the need to take effective action in the area of quality assurance in order to make the most of the existent savings potential. Transparency of treatment processes is a necessary condition for an economic assessment and for optimal resource allocation. PMID- 16362135 TI - [Usability of databases in health economics]. AB - In the health care system, there is increasing demand for health economic analyses to achieve cost reductions or increase efficiency. Apart from classical literature or study-based analyses in health economics, routinely obtained data may complement hitherto missing information for the general medical care situation. By analysing care structures and comparing applied or newly introduced technologies, results can be achieved faster and the significance for payers increased as compared to primary studies. Routinely obtained and potentially usable databases are frequently generated from surveys or from administrative data sources. In spite of clear limitations, these data are interesting in health economics based upon reflection of reality in utilization of services. They can contribute to improved understanding of general medical care and aid in identifying economically appropriate technologies and therapeutic strategies. PMID- 16362136 TI - [Biomechanical aspects of revision components for knee arthroplasty]. AB - Total knee arthroplasty (TKA) is one of the most frequent orthopaedic surgical procedures. Despite continuous improvements in the endoprostheses, instruments, and operative techniques, revision TKA has a rate of about 10% of overall TKA. In addition to the restoration of the periprosthetic bone stock and a precise alignment, the choice of an adequate implant, which meets the patient's specific requirements, has high impact on the outcome. The most significant differences between implants involve the degree of reconstructed joint area (uni-, bi-, tri compartimental) and the order of the constraining forces between the femoral and tibial component. Implants for revision TKA commonly range from un- or semiconstrained resurfacing implants to fully constrained hinged endoprostheses. In case of severe osseous, ligamentous, and/or muscular defects, special tumor endoprostheses or implants for arthrodesis might be an alternative option. PMID- 16362137 TI - [Revision surgery for knee prostheses using the DRG system]. AB - Due to an increasing life expectancy and earlier primary implantation of total knee replacements, the number of patients requiring revision surgery in Germany is increasing by 7% every year. These revision cases belong to the most treatment and cost intensive operations in joint replacement surgery. Presently, the description of these procedures in the German DRG system, which defines the financial reimbursement for the hospitals, is changing yearly with the development of new catalogues. The changes made from 2003 to 2005 are outlined in the following article. A correct depiction of the treatment and procedures required in such cases is a prerequisite for an adequate reimbursement. In the long-term, hospitals will only be able to offer such complex treatment forms if the financial compensation correctly reflects the costs incurred. PMID- 16362138 TI - [Perioperative pain therapy for knee endoprosthetics]. AB - Total knee arthroplasty (TKA) is associated with significant postoperative pain. Adequate analgesics and techniques are required for early mobilization, continuous passive motion and intensified physical therapy as well as for high quality postoperative analgesia.However, in the immediate postoperative setting the excessive nociceptive input can be blocked by using doses which are most frequently associated with adverse effects like dizziness, nausea and vomiting, sedation and risk of respiratory depression. The use of peripheral nerve blocks is recommended after orthopaedic surgery. After TKA, the continuous "3 in 1 nerve block" has been proven to be more effective than conventional patient controlled intravenous opioid therapy as well as than epidural analgesia accompanied by side effects. Postoperative analgesic techniques influence surgical outcome, duration of hospitalization and re-convalescence. The use of regional analgesia after TKA may initially lead to higher costs but it is counterbalanced by a reduction in morbidity and mortality, decrease in hospitalization, improved re-convalescence and a better functional outcome. PMID- 16362139 TI - [Reconstruction of the extensor tendons in revision total knee arthroplasty and tumor surgery]. AB - Reconstruction of the extensor mechanism in extended revision after total knee replacement and tumor surgery remains a clinically relevant problem. Due to large tibial bone defects with resection of the extensor insertion area, the specific problem of patella ligament refixation frequently arises. Several biological approaches and augmentation techniques have been published. Most of these are associated with a high rate of revision surgery because of failed replacement of the extensor mechanism and unsatisfactory functional outcome. Surgical reconstruction of these tendon defects is complicated by the difficulty of completely neutralizing tensional force across the repair. To overcome this problem, methods have been developed to reinforce the reconstruction with overlapping flaps; in addition, artificial materials are being increasingly used for tension neutralization. These artificial strips need special fixation mechanisms on the tibial component and specific technical modifications of the prosthesis. The present study gives an overview of reconstruction modalities of the extensor mechanism and provides an improved technology for better reconstruction by using artificial strips combined with specific modifications of the tibial component. PMID- 16362140 TI - [Axial correction in knee revision arthroplasty]. AB - Implant malalignment is a major cause for early loosening, increased wear, painful limitation of motion, and patient dissatisfaction in total knee arthroplasty. Validated diagnostic algorithms and a deeper understanding of the pathological mechanisms underlying functional deficits and pain resulting from malalignment explain the increasing number of revision operations on unloosened prostheses, which are now nearly as common as revisions for implant loosening. Common reasons are component malpositioning are a shifted joint line, or a non physiological patella position. The success of any revision procedure basically depends on: (1) correct component positioning, (2) equal and symmetrical flexion and extension gaps, (3) restoration of joint line, and (4) a physiological patella height. The adequate grade of implant constraint has to be determined intra-operatively. A higher loosening rate of constrained implants as well as increased wear and painful limitation of motion in case of instability have to be taken into account. In the present work, a diagnostic and therapeutic algorithm for malalignment of knee prostheses is presented. PMID- 16362141 TI - [Hip arthroscopy. Portal technique and arthroscopic anatomy]. AB - In hip arthroscopy everything revolves around entry points. Only with a precise entry technique are safety and efficiency guaranteed. Most pathological changes of the hip for which an arthroscopic intervention can be used are located in the central compartment: two lateral (anterolateral and posterolateral) and one anterior portal are the three standard portals allowing an optimal approach to practically all pathological changes in this compartment. Significant diseases and lesions can occur in the peripheral compartment which would be overlooked by arthroscopy of the central compartment. Two portals are needed for arthroscopy of the periparal compartment: the anterolateral, as well as one distally, converging on the ventral neck of the thigh. Hip arthroscopy is undergoing continual development, however, there are still major requirements for clarification of the pathology and pathomechanisms of injuries and diseases of the this joint. PMID- 16362143 TI - Acute pelvic organ prolapse in an 11-month-old infant. AB - There exist several reports of pelvic organ prolapse in high-risk neonates, but acute pelvic organ prolapse in a healthy infant has never been reported. In the current case, the parents first noted their 11-month-old infant with a mass protruding from the vagina after playing in a baby walker. Third-degree uterine prolapse was subsequently diagnosed after careful physical examination and pelvic ultrasonography. Under general anesthesia, the uterus was replaced after emptying the bladder. No sequelae were noted during 6 months of follow-up. PMID- 16362142 TI - The puzzle of overactive bladder: controversies, inconsistencies, and insights. AB - Overactive bladder (OAB) affects millions of individuals and may severely impair the quality of life of those affected. The contribution of human behavior to manifestations of this symptom complex remains poorly understood. Continued evolution of our understanding of the pathophysiology of OAB has identified contributory mechanisms, which in turn may open new therapeutic avenues. Recent improvements in drug delivery systems represent advances in the management of OAB. However, more complete symptom control with greater tolerability is desirable; this awaits the development of agents specific for newly emerging and as yet unidentified pathophysiologic pathways. Importantly, as understanding of outcomes assessment in OAB matures, refined assessments of disease severity, response to intervention, and patient preference should be possible. PMID- 16362145 TI - Differences in bone size and bone mass between black and white 10-year-old South African children. AB - INTRODUCTION: Black and white South Africans hail from vastly disparate cultural and socio-economic backgrounds the result of which exposes black children to numerous factors known to impact negatively on bone mass. Thus, we studied ethnic differences in bone size and bone mass between 476 10-year-old black and white South African girls and boys (black boys n=182, white boys n=72, black girls n=158, white girls n=64) who formed part of a longitudinal cohort of children born in Johannesburg, South Africa, during 1990. METHODS: Bone area (BA) and bone mineral content (BMC) were measured at the whole body, total hip, femoral neck, lumbar spine (L1-L4) and mid- and distal radii by dual energy X-ray absorptiometry (DXA). Vertebral heights and metacarpal indices were measured. Anthropometry, skeletal maturity and pubertal development were also assessed. RESULTS: After correction for height, weight, gender and puberty, black children had greater BMC at the femoral neck (P<0.0001), total hip (P<0.05) and mid-radius (P<0.001) than white children.. At the whole body, lumbar spine, and distal one third of the radius, there were no differences in BMC between black and white children after correction for differences in body size. After correction for height and puberty, vertebral heights were less in black children than white children, and cortical areas at the second metacarpal were greater in black children. CONCLUSION: These findings suggest that, at the femoral neck, total hip and mid-radius, these differences are not a result of differences in anthropometry, bone age or pubertal stage, or environmental factors but are most likely to result from genetic differences. PMID- 16362146 TI - Development of the Qualeffo-31, an osteoporosis-specific quality-of-life questionnaire. AB - INTRODUCTION: Vertebral deformities are a common consequence of osteoporosis and are known to decrease quality of life. The Qualeffo-41 is a quality-of-life questionnaire especially developed for measuring quality of life in patients with vertebral deformities. It consists of 41 questions arranged in five domains: pain, physical function, social function, general health perception, and mental function. The objectives of this study were: (1) to develop a shorter version of the Qualeffo-41 by removing redundant questions; and (2) to investigate the scale characteristics, reliability, and validity of this shorter version. METHODS: The study was performed using data from the Qualeffo validation study and the Multiple Outcomes of Raloxifene Evaluation (MORE) study. The analyses were performed in patients with vertebral deformities (n=579). Factor analysis on polychoric correlations and an item response theory (IRT) model, i.e., the generalized partial credit model (GPCM), were used to create a shorter version of Qualeffo-41. Using GPCM, scoring weights were computed for all items. RESULTS: Three items were removed from the data set because of too many missing values. Factor analysis identified three instead of five domains: (1) pain, (2) physical function, and (3) mental function. Five items had factor loadings <0.4 and were not included in the GPCM. After excluding several items, the domains pain (four items), physical function (18 items), and mental function (nine items) showed a good, reasonable, and excellent fit, respectively. This indicates that the mental function domain and the pain domain are more unidimensional than the physical function domain. All three domains showed a very high correlation (r > or =0.95) with the corresponding domains of the Qualeffo-41. CONCLUSIONS: Qualeffo-31 was developed, consisting of three domains with a reasonable to excellent fit to the GPCM. Although the fit to the GPCM supports the construct validity of the Qualeffo-31, validation in a new study should be performed before using it in practice. PMID- 16362144 TI - Changes in bone mass and bone turnover following tibial shaft fracture. AB - INTRODUCTION: Bone loss occurs in the regional bone following tibial shaft fracture. An earlier cross-sectional study showed that measurements made at the metaphyseal region of the tibia using peripheral quantitative computed tomography (pQCT) and the ultradistal region of the tibia using dual-energy X-ray absorptiometry (DXA) were the most responsive at monitoring this bone loss. Biochemical markers of bone turnover enable us to assess the activity of bone formation and resorption during fracture healing. The aim of this longitudinal study was to determine the pattern and distribution of bone loss and bone turnover following a tibial shaft fracture treated with either plaster cast or intramedullary nail. METHODS: Eighteen subjects underwent bone mass measurements using DXA at the tibia and hip and quantitative ultrasound (QUS) at the tibia and calcaneus of both limbs at 2 weeks, 8 weeks, 12 weeks and 24 weeks following fracture, with hip and tibia DXA measurements also performed at 52 weeks. Nine of the patients treated with plaster cast had pQCT measurements at the tibia at 24 weeks. We measured three bone formation markers, bone alkaline phosphatase (bone ALP), osteocalcin (OC) and procollagen type 1 N-terminal peptide (PINP), a marker of bone resorption, serum C-telopeptides of type 1 collagen (beta-CTX) and a marker of collagen III turnover, procollagen type III N-terminal peptide (PIIINP) at 1 day, 3 days and 7 days and at 2, 4, 8, 12, 16 and 24 weeks following fracture. The greatest bone losses were observed at the ultradistal region of the tibia using DXA (28%, p <0.001) and the metaphyseal region of the tibia using pQCT (26-31%, p <0.001) at 24 weeks. In the hip, the greatest loss was in the trochanter region at 24 weeks (10%, p <0.001). The greatest loss at the calcaneus measured using QUS was for broadband ultrasound attenuation (BUA) measured using CUBA Clinical at 24 weeks (13%, p =0.01). RESULTS: At 1 year, there was a small recovery in bone loss (ultradistal tibia DXA, 20%, p <0.01; trochanter DXA 9%, p <0.001). Bone turnover increased following fracture (PINP +72+/-21%, p <0.0001, bone ALP +199+/-22%, p =0.004, beta-CTX +105+/-23%, p <0.0001, all at 24 weeks). There was a smaller +33+/-10% increase in osteocalcin at 24 weeks. PIIINP concentration peaked at week 8 (+57+/-9%, p <0.0001). The bone resorption marker beta-CTX showed an earlier rise (week 2, 139+/-33%) than the bone formation markers. CONCLUSIONS: We conclude that: (1) bone loss following tibial shaft fracture occurs both proximal and distal to the fracture; (2) the decreased BMD is largest for trabecular bone in the tibia with similar measurements using DXA and pQCT; (3) there is limited recovery of bone lost at the hip and tibia at 1 year; (4) tibial speed of sound (SOS) demonstrated a greater decrease than calcaneal SOS when comparing z -scores; (5) BUA is the QUS variable that shows the biggest decrease of bone mass at the calcaneus; (6) increase in bone turnover occurs following fracture with an earlier increase in bone resorption markers and a later rise in bone formation markers. PMID- 16362147 TI - Alendronate for osteoporosis in men with androgen-repleted hypogonadism. AB - Male hypogonadism is associated with low bone mineral density (BMD) and an increased risk of fractures. Testosterone replacement therapy improves BMD in young hypogonadal men. This effect is milder in older patients, who are at greater risk for fractures. We studied the effects of alendronate or placebo on BMD in 22 osteoporotic men, 29-69 years of age (mean, 50.2+/-11.2 years) with long-standing hypogonadism, receiving standard testosterone replacement treatment. Alendronate 10 mg daily (n=11) increased lumbar-spine BMD by 6.0 and 8.4% at 6 and 12 months, respectively, compared with -0.5% at 6 months and +3.3% at 12 months in the placebo group (n=11; P<0.005). Alendronate also increased mean femoral-neck BMD by 1.9% after 1 year, compared to a 1.4% decrease with placebo (P<0.005), and increased the total body bone mineral content by 4.4%, compared to a 0.6% decrease with placebo (P=0.07). After 6 months alendronate suppressed urinary deoxypyridinoline by 50% (P<0.005), compared to a 24% decrease in the placebo group. Both the alendronate and placebo groups continued with alendronate 70 mg once weekly for the following 2 years. Lumbar-spine BMD during this open-label study phase did not change significantly in the group originally treated with alendronate, but continued to increase in the placebo-alendronate group by 5.4, 6.5, and 6.2% after 18 (6 months of alendronate), 24 and 36 months, respectively (P<0.05). Femoral-neck BMD continued to increase in both groups receiving active therapy; in the alendronate-alendronate group by 3.7, 2.7, and 5.2% after 18, 24, and 36 months, respectively (P=0.01), and in the placebo alendronate group by 0.7 and 1.9% at 24 (first 12 months of alendronate) and 36 months, respectively (P<0.05). Our results support the long-term administration of alendronate along with testosterone replacement to men with hypogonadism induced osteoporosis. PMID- 16362151 TI - Moving with the flow: what transport laws reveal about cell division and expansion. AB - This material was presented as a keynote talk for the symposium, "Crosstalk between cell division and expansion," organized by G.T.S. Beemster and H. Tsukaya at the International Botanical Congress, Vienna in July, 2005. The review focuses on the utility of continuity equations to understand relationships among cell size, division and expansion; insights from Lagrangian or cell-specific descriptions of developmental variables; and a growth-diffusion equation to show effects of root growth zones on the surrounding soil. PMID- 16362149 TI - Usefulness of 3'-[F-18]fluoro-3'-deoxythymidine with positron emission tomography in predicting breast cancer response to therapy. AB - OBJECTIVE: The usefulness of 2-deoxy-2-[F-18]fluoro-D-glucose (FDG)-positron emission tomography (PET) in monitoring breast cancer response to chemotherapy has previously been reported. Elevated uptake of FDG by treated tumors can persist however, particularly in the early period after treatment is initiated. 3'-[F-18]Fluoro-3'-deoxythymidine (FLT) has been developed as a marker for cellular proliferation and, in principle, could be a more accurate predictor of the long-term effect of chemotherapy on tumor viability. We examined side-by-side FDG and FLT imaging for monitoring and predicting tumor response to chemotherapy. METHODS: Fourteen patients with newly diagnosed primary or metastatic breast cancer, who were about to commence a new pharmacologic treatment regimen, were prospectively studied. Dynamic 3-D PET imaging of uptake into a field of view centered over tumor began immediately after administration of FDG or FLT (150 MBq). After 45 minutes of dynamic acquisition, a clinically standard whole-body PET scan was acquired. Patients were scanned with both tracers on two separate days within one week of each other (1) before beginning treatment, (2) two weeks following the end of the first cycle of the new regimen, and (3) following the final cycle of that regimen, or one year after the initial PET scans, whichever came first. (Median and mean times of early scans were 5.0 and 6.6 weeks after treatment initiation; median and mean times for late scans were 26.0 and 30.6 weeks after treatment initiation.) Scan data were analyzed on both tumor-by-tumor and patient-by-patient bases, and compared to each patient's clinical course. RESULTS: Mean change in FLT uptake in primary and metastatic tumors after the first course of chemotherapy showed a significant correlation with late (av. interval 5.8 months) changes in CA27.29 tumor marker levels (r = 0.79, P = 0.001). When comparing changes in tracer uptake after one chemotherapy course versus late changes in tumor size as measured by CT scans, FLT was again a good predictor of eventual tumor response (r = 0.74, P = 0.01). Tumor uptake of FLT was near-maximal by 10 minutes after injection. The time frame five to 10 minutes postinjection of FLT produced standardized uptake value (SUV) values highly correlated with SUV values obtained after 45-minute uptake (r = 0.83, P < 0.0001), and changes in these early SUVs after the first course of chemotherapy correlated with late changes in CA27.29 (r = 0.93, P = 0.003). CONCLUSION: A 10 minute FLT-PET scan acquired two weeks after the end of the first course of chemotherapy is useful for predicting longer-term efficacy of chemotherapy regimens for women with breast cancer. PMID- 16362152 TI - Pathogenesis of vascular calcification in dialysis patients. AB - Soft-tissue and vascular calcification are highly prevalent in end-stage renal disease (ESRD). Vascular calcifications manifest as both medial and intimal calcification of arteries and are a hallmark of the accelerated atherosclerosis observed in uremia. The nature of vascular calcification is progressive, and is associated with arterial stiffness and increased cardiovascular mortality. Age, duration of dialysis, and diabetes mellitus are clear determinants of the severity of vascular calcification; however, more recently novel insights into the pathomechanisms of unwanted calcification processes have been gained. Disturbances of mineral metabolism such as hyperphosphatemia and hypercalcemia appear to contribute to progressive calcification, not only by passive precipitation but by actively inducing changes in vascular smooth muscle cell behavior toward an osteoblast-like phenotype. Specific calcium-regulatory proteins may act locally or systemically as calcification inhibitors. Dysregulations of calcification inhibitors, including fetuin-A, matrix Gla protein, osteoprotegerin, and pyrophosphates may also be pathophysiologically relevant factors in the context of uremic extraosseous calcification. In this context, low serum fetuin-A levels were recently found to be associated with increased mortality in cohorts of dialysis patients. This overview intends to summarize current knowledge of the scientific concepts involved in the pathogenesis of extraosseous calcification in ESRD. PMID- 16362150 TI - Use of positron emission tomography to target prostate cancer gene therapy by oncolytic herpes simplex virus. AB - Herpes simplex virus (HSV) oncolytic gene therapy is a promising treatment modality against cancer. We have demonstrated that androgen-induced cellular changes enhance oncolytic viral replication and improve efficacy in the treatment of androgen-dependent prostate cancer cell line. Imaging of changes in 2-deoxy-2 [F-18]fluoro-D-glucose (FDG) uptake by positron emission tomography (PET) is a sensitive method of detecting altered cellular metabolism involved in cancer therapy. We therefore hypothesized that FDG-PET can predict tumor response to oncolytic HSV therapy. In this study, androgen increased cell kill (74%) in vitro and enhanced viral yield (2.4-fold) in vivo following HSV therapy. This enhanced efficacy was predicted by high FDG accumulation in intact animals compared to low FDG uptake following orchiectomy (p = 0.002). This proof-of-concept study provides the mechanistic basis for selecting patients for targeted oncolytic viral therapy by means of a noninvasive molecular imaging method in the treatment of prostate cancer. PMID- 16362153 TI - Novel therapeutic approaches for progressive renal disorders by targeting glomerular component mesangial and endothelial cells. PMID- 16362155 TI - Identification of immature podocyte specific antigen using retrovirus-mediated gene transfer and cell sorting. AB - BACKGROUND: For identifying an antigen recognizable by a monoclonal antibody (mAb), the cDNA cloning method using expression cDNA library has become increasingly popular. For analysis of the mAb clone 2E3, which recognizes the cell membrane surface of immature podocytes, we developed an expression cloning strategy that identified the targeted antigen using retrovirus-mediated gene transfer and enrichment by cell sorting. METHODS: In this experiment, the NIH3T3 cell line was infected by a retrovirally amplified cDNA library derived from the same fetal murine kidneys that provided the immunized antigen. Infected NIH3T3 staining for mAb clone 2E3 was concentrated by cell sorting, followed by identification of the integrated gene. RESULTS: The infected cells reacted with mAb were highly enriched by two rounds of cell sorting. As a result of sequencing the inserted gene from the enriched cells, we isolated the low-affinity nerve growth factor receptor (NGFR) gene. Furthermore, the NIH3T3 cell line that was enforced by recovery of NGFR cDNA reacted with mAb clone 2E3. CONCLUSIONS: Monoclonal antibodies recognizing cell membrane surface molecules are essential tools for immunohistochemistry and flow cytometry analysis, and the application of a combination of retrovirus-mediated gene transfer and cell sorting is beneficial for identifying the targeted antigen. PMID- 16362154 TI - ADPKD: molecular characterization and quest for treatment. AB - Autosomal-dominant polycystic kidney disease (ADPKD) is a common hereditary disease that features multiple cystogenesis in various organs and vascular defects. The genes responsible for ADPKD, PKD1, and PKD2 have been identified, and the pathological processes of the disease are becoming clearer. This review focuses on recent findings about the molecular and cellular biology of ADPKD, and especially on PKD1. PKD1 and its product, polycystin-1, play pivotal roles in cellular differentiation because they regulate the cell cycle and because polycystin-1 is a component of adherens junctions. A possible link between polycystin-1 and PPARgamma is discussed. The extraordinarily fast research progress in this area in the last decade has now reached a stage where the development of a remedy for ADPKD might become possible in the near future. PMID- 16362156 TI - Monocyte chemoattractant protein-1 A-2518G gene polymorphism and renal survival of Japanese patients with immunoglobulin A nephropathy. AB - BACKGROUND: Monocyte chemoattractant protein (MCP)-1 is closely related to the pathogenesis of the progression of various chronic renal diseases, including IgA nephropathy (IgAN), through its chemoattractant effect on macrophages. However, the correlation of MCP-1 gene polymorphism with the long-term prognosis of Japanese patients with IgAN has not been clearly determined yet. METHODS: We investigated 277 Japanese patients diagnosed with IgAN based on renal biopsy to clarify the association between the progression of IgAN and MCP-1 gene polymorphism at position A-2518G, which regulates the transcription of the MCP-1 gene. RESULTS: The incidence of endstage renal disease was significantly higher in patients with the AA genotype (47.1%) compared to those with the AG (24.1%) or GG (27.4%) genotype (P = 0.024). Moreover, Kaplan-Meier analysis revealed that the AA genotype significantly facilitated the progression of renal disease (log rank; P = 0.0029), and Cox proportional hazards regression model analysis showed that the AA genotype represented a 2.058-fold risk for the progression of renal disease (P = 0.026) compared to the AG/GG genotype. However, when the patients were treated with angiotensin-converting enzyme inhibitor and/or angiotensin receptor blocker, or corticosteroid, homozygosity for the -2518A allele was not associated with a higher rate of incidence of endstage renal disease. Serum MCP-1 levels were higher although not significantly so, in the patients with IgAN possessing the AA genotype. CONCLUSIONS: The AA genotype at MCP-1 -2518 was an independent risk factor for the progression of renal disease in Japanese patients with IgAN, and was closely associated with renal survival. PMID- 16362157 TI - A useful new classification of dysmorphic urinary erythrocytes. AB - BACKGROUND: Among dysmorphic urinary erythrocytes (D cells), G1 cells or doughnut shaped erythrocytes with one or more blebs are considered to be reliable markers for glomerular diseases. However, although there are many D cells with cytoplasmic color loss and without blebs in the urinary sediment, the significance of these cells is not clear. In this study, we devised a classification system for D cells and examined the relation between these cell types and urinalysis data. METHODS: We classified D cells into three types (D1, D2, and D3 cells): D1 cells showed a ring-like shape and severe loss of cytoplasmic color with protrusions or blebs; D2 cells showed a doughnut-like shape and moderate cytoplasmic color loss with protrusions or blebs; and D3 cells showed a doughnut-like shape and mild cytoplasmic color loss without protrusions or blebs. We calculated the numbers of D cells of each type in 45 patients with glomerular diseases and in 303 general outpatients. This was done by bright-field microscopy modified for the analysis of urinary sediment, and we also examined the significance of these cell types. RESULTS: In the 45 patients with glomerular diseases, the numbers of D1, D2, and D3 cells correlated with urine levels of proteinuria and hematuria and numbers of cellular and fatty casts. Numbers of D1 and D2 cells correlated with urine concentrations of albumin and N-acetyl-beta-D glucosaminidase, and the proportions of D1 and D2 cells in D cells increased with the activity of glomerular diseases classified by urinalysis data. Only the number of D1 cells correlated with the urine concentration of potassium, which may increase in hemolysis. In the 303 outpatients, the sensitivity of D3 cells and D1 and/or D2 cells (G1 cells) was 73% and 46%, respectively, for the detection of glomerular diseases and the specificity was 93% and 99%, respectively. CONCLUSIONS: These data indicate that the D3 cell is a sensitive marker for glomerular diseases, and that D1 and/or D2 cells are markers for severe glomerular diseases. PMID- 16362158 TI - Imbalance between production and scavenging of hydroxyl radicals in patients maintained on hemodialysis. AB - BACKGROUND: Reactive oxygen species are as being related to the pathophysiology of endstage renal disease (ESRD). We measured the plasma hydroxyl radical (.OH) producing ability and .OH-scavenging activity in patients with ESRD to clarify the pathophysiological states involved. METHODS: We used electron spin resonance to measure plasma N-t-butyl-alpha-phenylnitron radical spin adduct (pPBN rsa) as .OH-producing ability and plasma 3,3,5,5-tetramethyl-1-pyrroline-N-oxide radical spin adduct (pM4PO rsa) as .OH-scavenging activity. Oxidative injuries were evaluated by determining oxidised low-density lipoprotein (Ox-LDL). RESULTS: The pPBN rsa of the ESRD patients was lower than that of the controls (1.83 vs 2.94 micromol/g protein). The pM4PO rsa of the ESRD patients was higher than that of the controls (3.85 vs 3.15 mmol L: -ascorbic acid 2-[3,4-dihydro-2,5,7,8 tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1-benzopyran-6-yl hydrogen phosphate] potassium salt (EPC-K1)/g protein). The pPBN rsa and pM4PO rsa were correlated, both in the ESRD patients and in the controls (r = 0.47 and r = 0.53). Ox-LDL was correlated with hemodialysis (HD) duration (r = 0.49) and was negatively correlated with pPBN rsa (r = -0.54), which indicates that oxidative stress was increased as HD therapy was prolonged and suppressed pPBN rsa. CONCLUSIONS: There was an imbalance between .OH-producing ability and .OH-scavenging activity, in the ESRD patients, and this may be responsible for compromising the health of ESRD patients. PMID- 16362159 TI - Restoration of peritoneal integrity after withdrawal of peritoneal dialysis: characteristic features of the patients at risk of encapsulating peritoneal sclerosis. AB - BACKGROUND: The epidemiological characteristics of encapsulating peritoneal sclerosis (EPS), such as its high incidence in patients with long-term peritoneal dialysis (PD) treatment, and the onset of EPS after patients are switched to hemodialysis (HD) may indicate an activated pathological process after PD withdrawal, especially in long-term PD patients. Accordingly, we aimed to observe changes in peritoneal function after the stoppage of PD, and to clarify the characteristic features of the patients at risk of EPS. METHODS: Thirty-three patients who were switched from continuous ambulatory peritoneal dialysis (CAPD) to HD were enrolled in this trial. Changes in the dialysate/plasma creatinine (D/P Cr) and CA125 levels in the effluent of the peritoneal equilibration test were observed for 6 months. Furthermore, each patient was followed-up for 36 months after PD withdrawal to monitor for the development of EPS. RESULTS: D/P Cr decreased significantly, while CA125 levels tended to increase. Nine patients developed EPS during the follow-up period and they specifically showed significant increases of D/P Cr levels and significantly lower levels of CA125 at PD withdrawal. The accumulation of high transporters in the EPS group at 0 and 6 months after PD withdrawal was significant. CONCLUSIONS: Peritoneal recovery may take place after withdrawal from PD treatment and such recover indicated by improvement of transport states and a rise of the CA125 level. The present study revealed that a high-transport state and lack of increase of CA125 in the effluent were associated with EPS development after PD withdrawal. This may suggest that the lack of peritoneal recovery after PD withdrawal is predictive for EPS development. PMID- 16362160 TI - Nephronophthisis in two siblings. AB - We describe here two sisters with nephronophthisis, which was not detected until the development of endstage renal failure. Twenty- and 15-year-old female siblings were admitted to our hospital for further examination of renal dysfunction. No urinalysis abnormalities had been found in yearly health checks in either patient. The serum creatinine level was 7.2 mg/dl in case 1 (the 20 year-old) and 6.4 mg/dl in case 2. Medical history, physical findings, and laboratory tests showed no evidence of urinary tract infection, use of any drugs, arthritis, or skin eruptions. To identify the cause of the renal failure, open left renal biopsies were performed in both patients. Histopathological findings were very similar in the two patients and included marked tubular and interstitial changes (tubular dilatation, focal tubular atrophy, interstitial fibrosis, and infiltration of mononuclear cells). The glomeruli were devoid of mesangial proliferation, mesangial expansion, and adhesion of Bowman's capsule. Based on the clinical and pathological findings, the final diagnosis was nephronophthisis in both patients. It is important to remember that some progressive renal diseases, including nephronophthisis, cannot be detected even by annual urinary screening tests, which are widely performed in Japan. PMID- 16362161 TI - Extensive pancreatic necrosis in microscopic polyangiitis. AB - We encountered an 84-year-old woman with microscopic polyangiitis who was found to have pancreatitis on autopsy. The patient was admitted to Yamanashi University Hospital because of fever and progressive renal failure. She was diagnosed with anti-myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody (ANCA)-related microscopic polyangiitis (MPA) and was treated successfully with prednisolone pulse therapy. Two months later, she was found unconscious at home and was transferred to hospital, where she died of cardiac arrest after 6 days. Autopsy revealed systemic vasculitis with fibrinoid necrosis (with the most severe form found in the pancreas), interstitial pneumonia, and crescentic glomerulonephritis. A review of the literature revealed that pancreatic involvement in vasculitis, although rare, is one of the complications of MPA; however, the present study is the first report to focus on the pancreatic involvement of MPA. We recommend that nephrologists consider the possibility of pancreatic involvement in this disease. PMID- 16362162 TI - Cimetidine-induced tubulointerstitial nephritis with both MPO-ANCA and PR3-ANCA. AB - We describe a 75-year-old man with tubulointerstitial nephritis (TIN) with myeloperoxidase (MPO)-antineutrophil antibody (ANCA) and proteinase-3 (PR3)-ANCA. He had a slight fever and eruption with itching after taking cimetidine (prescribed after gastrectomy for gastric cancer) and he was admitted to a nearby hospital. There, he showed proteinuria, serum creatinine (sCr) of 2.9 mg/dl, and creatinine clearance (Ccr) of 44 ml/min per 1.73 m2. His MPO-ANCA titer was 267 EU, and PR3-ANCA titer was 112 EU. Abnormal concentrations in bilateral kidneys were found by gallium scintigraphy. For these reasons, he was transferred to our hospital. Percutaneous renal biopsy was performed after admission. Severe tubular atrophy, mild interstitial fibrosis, and severe mononuclear cell infiltration of the interstitium were noted. Drug-induced renal impairment was suspected, and cimetidine administration was withdrawn. Lymphocyte stimulation tests (DLSTs) were performed. The cimetidine titer was positive, at 2,537 cpm. After the withdrawal of cimetidine, the PR3-ANCA titer was reduced gradually, and, next, the MPO-ANCA titer was also reduced. The sCr level was reduced to 1.2 mg/dl. In summary, we report herein the first case of cimetidine-induced TIN associated with both MPO-ANCA and PR3-ANCA. PMID- 16362163 TI - Catastrophic antiphospholipid antibody syndrome following initiation of hemodialysis. AB - Antiphospholipid syndrome (APS) is associated with arterial and venous thrombosis, pregnancy morbidity, and thrombocytopenia. Some APS patients develop rapid and disseminated microthrombosis and are known as having catastrophic APS or CAPS. We document here a case of CAPS in a patient who presented with various clinical symptoms and serious abnormalities of blood coagulation following initiation of hemodialysis after bilateral nephrectomy due to renal cancer. The patient developed multiple organ symptoms, including melena, visual disturbances, skin eruptions, lymph node swelling, urinary tract bleeding, backache, arteriovenous fistula occlusion, and chest pain. Based on the clinical course and serological and histological examinations, a diagnosis of CAPS was established. The patient recovered by following intensive anticoagulation and steroid therapy. Although CAPS is rare, once symptoms develop the condition deteriorates rapidly. Because of the associated high mortality, early diagnosis and prompt treatment are necessary. PMID- 16362164 TI - Nephronophthisis: diagnostic difficulties and recent advances in molecular genetic diagnostics. PMID- 16362165 TI - Renal failure due to djenkolism: an appraisal of previously reported Thai cases. PMID- 16362166 TI - Response categories and anger measurement: do fewer categories result in poorer measurement?: development of the DAR5. AB - BACKGROUND: Anger is a key long-term outcome from trauma exposure, regardless of trauma type, and it is implicated as a moderator of response to treatment. It therefore seems important that anger is assessed in both epidemiological studies of trauma sequelae and in intervention evaluation research. This study explored the measurement properties of a recently investigated anger scale, the Dimensions of Anger Reactions (DAR) Scale. In our previous study, the DAR was found to be a measure of trait anger, but although brief, the nine response categories per item may have confused respondents, suggesting fewer response categories may work equally well. Additionally, our previous analysis suggested there were two redundant items within the DAR. METHODS: Three samples of Australian veterans were used to investigate the psychometric properties associated with alterations to the response categories of the DAR; veterans who participated in the DAR validation study, those participating in group therapy programmes for post traumatic stress disorder, and veterans participating in lifestyle programmes. Item response theory analysis was used to explore the internal properties of competing DAR models, and models were assessed against external criteria. RESULTS: The results showed that the number of item responses in the DAR exceeded channel capacity, and that response bias occurred in the second half of the instrument. We hypothesized that this was due to respondents not discriminating among the many response categories. Based on a modelling exercise in which we reduced the number of DAR items from 7 to 5 and the number of response categories from 9 to 5, validation tests showed that there was no loss of sensitivity, reliability or validity. To avoid confusion with the DAR, we have referred to the revised version of the DAR as the DAR5. CONCLUSIONS: We conclude that the DAR5, which abbreviates the original DAR to half its original length, has similar psychometric properties and is therefore to be preferred especially for use with persons who are under stress, cognitively impaired or less mature. The study findings regarding the optimum number of response categories have implications for the development of other instruments. PMID- 16362167 TI - Core features of repeated suicidal behaviour: a long-term follow-up after suicide attempts in a low-suicide-incidence population. AB - BACKGROUND: The aim of this article was to study repeated suicidal behaviour in a low-incidence population to elucidate robust risk factors. METHODS: A cohort of first-ever suicide attempters from 1960 to 1982 on the Faroe Islands was followed up for a minimum of 20 years. The cohort was initially characterized in psychiatric and social terms. RESULTS: The incidence of suicidal behaviour for the cohort years (age 15 years and older) was 37.9 per 100,000 per year (95% confidence interval 31.5-45.1). It was associated, as expected, with gender, age, residence, marital status, occupation, diagnosis, previous psychiatric admission, alcohol intoxication and the method and planning of the act. Factors of the index episode predicting repetition at 5 years were gestures and alcohol intoxication and at 20 years were physical methods, suicide letter and alcohol intoxication. CONCLUSIONS: Alcohol intoxication and the level of determination behind the suicide attempt emerge as targets for prevention. Alcohol intoxication at the initial episode seems to be a strong long-term as well as short-term risk factor. PMID- 16362168 TI - Suicide mortality in South Africa: a city-level comparison across socio demographic groups. AB - BACKGROUND: South Africa offers a valuable study environment to assess the impact of political and social changes on health. Increasing urbanisation has led to a focus on city development and its effect on its residents. The study assesses whether sex- and race-specific suicide rates differ across six of South Africa's major cities. Regional differences may reflect varying contextual and individual underlying mechanisms. Those need to be understood for appropriately targeted prevention efforts. METHODS: Suicide mortality data (n=4,946) for the six cities between 2001 and 2003 were used to assess whether there are differences in rates for age-standardised race and sex groups across cities and whether these differences are constant across cities. The overall age-standardised rates for the six cities was 25.3/100,000 for men and 5.6/100,000 for women. RESULTS: Suicide rates are most often highest among whites and men, but both the magnitude and distribution of suicide vary considerably for different race and sex groups within and across cities. There is a strong association between the method of suicide used and the city. CONCLUSIONS: Despite some common trends in the suicide profile across South African cities, city does matter for the magnitude and distribution of suicide mortality across race and sex groups and for the method of suicide used. The need for locally based suicide research investigating possible reasons for these differences is highlighted. PMID- 16362169 TI - Imminent pulmonary embolism: a fatal mobile right atrial thrombus. AB - Right sided heart thrombi are infrequent and if they are mobile they may cause serious morbidity and mortality due to massive pulmonary embolism or paradoxical embolism. Malignancies are one of the important etiological factors for right heart thrombi. A patient with operated but recurrent ovarian carcinoma, presented with symptoms of heart failure was admitted to oncology department. Rapidly progressing dyspnea and a pre-syncope attack required consultation of a cardiologist and echocardiography revealed a mobile thrombus in the right atrium. Urgent open heart surgery was decided but imminent massive pulmonary embolism complicated the case leading to irreversible cardiogenic shock. By means of the presented case this paper overviews etiological factors and treatment options for right sided heart thrombi. PMID- 16362170 TI - A large calcified saccular aneurysm in a patient with aortic coarctation. AB - Coarctation of the aorta (CA) accounts for 5% of all congenital heart disease. One of the most feared complications of coarctation of the aorta is the development of saccular aortic aneurysm. In this paper, we described a 20-year old man with coarctation of the aorta in association with a large calcified saccular thoracic aneurysm just distal to the coarcted segment detected both in aortography and magnetic resonance angiography (MRA). PMID- 16362171 TI - In vivo variability in quantitative coronary ultrasound and tissue characterization measurements with mechanical and phased-array catheters. AB - BACKGROUND: Both mechanical and phased-array catheters are used in clinical trials to assess quantitative parameters. Only limited evaluation of the in vivo agreement of volumetrical measurements between such systems has been performed, despite the fact that such information is essential for the conduction of atherosclerosis regression trials. METHODS AND RESULTS: We prospectively evaluated the agreement in morphometric measurements and intravascular ultrasound (IVUS)-based plaque characterization between a 40 MHz rotating transducer (3.2 F Atlantis, Boston Scientific Corp.) and a 20 MHz phased-array catheter (2.9 F Eagle Eye, Volcano Therapeutics, Rancho Cordova, California) in 16 patients. Lumen (7.3+/-2.0 mm(2) vs. 6.7+/-1.8 mm(2), p=0.001) and vessel (11.8+/-3.3 mm(2) vs. 11.0+/-2.9 mm(2), p=0.02) cross-sectional areas (CSA) were significantly greater with the 20 MHz system. Plaque CSA measurements showed no significant difference between systems (4.4+/-2.3 mm(2) vs. 4.4+/-2.1). The relative differences were less than 10% for the three variables. On IVUS-based tissue characterization (13 patients), calculated percentage hypoechogenic volume was significantly higher for the 20 MHz system (96.7+/-2.38 vs. 88.4+/-5.53, p<0.0001). CONCLUSIONS: Quantitative IVUS analyses display significant catheter type-dependent variability. It is unclear whether the variability reflects overestimation of measurements with the phased-array or underestimation with the mechanical system. Although plaque burden measurements did not differ significantly between systems, it appears prudent to recommend the use of a single system for progression/regression studies. PMID- 16362172 TI - Characterization of alterations in diabetic myocardial tissue using high resolution MRI. AB - Cardiovascular complications, including diabetic cardiomyopathy, are the major cause of fatalities in diabetes. Diabetic cardiomyopathy is expressed in part through fibrosis and left ventricular hypertrophy, increasing myocardial stiffness leading to heart failure. In order to search for curative interventions, precise evaluation of the diabetic heart pathology is extremely important. Magnetic resonance imaging (MRI) is ideally suited for the assessment of heart disorders due to its high resolution, three-dimensional properties and dimensional accuracy. In this study streptozotocin injected Sprague-Dawley rats were used as a model of type 1 diabetes to characterize abnormalities in the diabetic left ventricle (LV). High resolution MRI using a 9.4 T horizontal bore scanner was performed on control and 7 weeks diabetic rats. In the diabetic rats as compared to controls, we found increased LV wall volume to body weight ratio, suggestive of LV hypertrophy; increased LV wall mean pixel intensity, and decreased T2 relaxation time, both suggestive of changes in the diabetic tissue properties, perhaps due to presence of fibrosis which was detected through increase in the collagen fractional area. In addition, changes in the LV cavity area were observed and quantified in post-mortem diabetic hearts indicative of stiffer and less resilient LV myocardial tissue with diabetes. Together the data suggest that LV hypertrophy and fibrosis may be a major factor underlying structural and functional abnormalities in the diabetic heart, and MRI is a valuable tool to non-invasively monitor the pathological changes in diabetic cardiomyopathy. PMID- 16362173 TI - A contractile left ventricular diverticulum. AB - We present the first case report of left ventricle muscular diverticulum which stands in front of the right ventricle in an asymptomatic 20-year-old male. The diverticulum has a connection with the left ventricular outflow truct, and contracts simultaneously with the left ventricle. The patient has been included in our congenital anomaly follow-up program. PMID- 16362174 TI - Double right coronary artery: review of literature. AB - Anomalies of the coronary artery are often asymptomatic and uncommon. We report an extremely rare coronary artery anomaly, a double right coronary artery. A 50 year-old male patient was hospitalized with the diagnosis of unstable angina pectoris. Right coronary artery (RCA) injection showed filling of two separately originating RCAs, coursing towards the right atrio-ventricular groove. Then they give a marginal branch and a septal artery, terminated by giving off the posterior descending artery (PDA) in posterior interventricular groove. Coronary anomalies should be recognized to avoid problems during coronary intervention and cardiac surgery. PMID- 16362176 TI - Extradural spinal arachnoid cysts associated with spina bifida occulta. AB - INTRODUCTION: Spinal extradural arachnoid cysts are an uncommon cause of neural compression in children. Even more uncommon is the association of such cysts with spina bifida occulta. MATERIAL: Two girls, 12 and 8-years-old, presented with left leg pain, deteriorating gait, clinical signs of left L5 and S1 root compression, without bladder or bowel symptoms. The first patient had left foot drop. The second patient had muscle wasting and smaller left foot with pes cavus. Radiographs showed spina bifida occulta of S1 in both. MRI revealed an extradural cyst at the S1 level, indenting the thecal sac and the L5 and S1 roots. At operation in both patients a large arachnoid cyst arising from a small dural defect in the axilla of the left S1 root was compressing and displacing it and the dural sac. It was removed and the defect was repaired. The first patient improved with complete recovery of the foot drop. An MRI at 12 months showed no cyst recurrence. The second patient made good recovery initially, but at 10 months developed recurrent symptoms. An MRI scan showed recurrence of the cyst with root compression. On repeat exploration a different dural defect was identified in a more anterior position and was repaired. DISCUSSION: The coexistence of extradural arachnoid cyst and corresponding bifid spinal segment has not been described previously. It raises the suspicion that the dural defect giving rise to the arachnoid cyst may be due to segmental dural dysgenesis in the context of the dysrhaphic neuroectodermal malformation. PMID- 16362175 TI - Rerupture following endovascular treatment for dissecting aneurysm of distal anterior inferior cerebellar artery with parent artery preservation: retreatment by parent artery occlusion with Guglielmi detachable coils. AB - Distal anterior inferior cerebellar artery (AICA) aneurysms are rare and most cases have been treated surgically by clipping, wrapping or trapping. We recently treated this 20-year-old male patient by an endovascular technique. At first, he was treated by intra-aneurysmal embolisation with parent artery preservation. But he presented with rerupture 1 month after embolisation. Follow-up angiography revealed the regrowth of the aneurysm, which was considered as a dissecting aneurysm. We performed occlusion of the AICA just proximal to the aneurysm to prevent fatal rebleeding. He gradually improved and his level of consciousness fully recovered. At 2 year follow up, he had no neurological deficits. We suggest that embolisation of distal AICA aneurysm with parent artery occlusion may be safe and a simple method in the treatment of distal AICA aneurysms. PMID- 16362177 TI - Intracranial retrograde dissemination in filum terminale myxopapillary ependymomas. AB - Myxopapillary ependymomas (ME) are considered benign tumours (WHO grade I) of the central nervous system with long term survival rates and a tendency to local recurrence. However an aggressive course has occasionally been described, leading to CSF dissemination and even systemic metastases. We describe the case of a 23 year-old man diagnosed with intracranial subarachnoid dissemination of a filum terminale ME three years after the initial diagnosis. We have performed a careful review of the literature on CSF dissemination in ME and finally propose treatment of these cases. PMID- 16362179 TI - DDAVP use in patients undergoing transsphenoidal surgery for pituitary adenomas. AB - BACKGROUND: Disorders of fluid and sodium regulation, often termed "diabetes insipidus," are a frequent occurrence following surgery for pituitary adenomas. The present study was undertaken to identify the incidence of diabetes insipidus after pituitary surgery and its associated factors. METHODS: A retrospective review of the medical records 300 patients who underwent transsphenoidal surgery for pituitary adenoma was undertaken. Information regarding patient gender, perioperative serum sodium levels and urinary output volumes, tumor size, previous pituitary surgery, tumor subtype, and the use of DDAVP was gathered. A multivariate statistical analysis was performed. FINDINGS: Follow-up data were available on 288 patients. During the inpatient postoperative hospital stay, DDAVP was administered to 19% of all patients and 16% of patients not taking DDAVP preoperatively. Of patients with normal fluid/sodium regulation preoperatively, DDAVP was prescribed for 9% at discharge and 4% at 6 weeks postoperatively. Only 1.4% of patients were taking vasopressing replacement at the time of last follow-up. Significant correlations were found between gender, previous surgery, serum sodium levels, and urine volumes at various time points. Immunohistochemical type of tumor and tumor size were not related to DDAVP requirement. CONCLUSIONS: Transient hypotonic polyuria is frequently encountered after pituitary surgery. However, only a small number of patients will develop a long-term requiring for ongoing medical treatment. Previous surgery, female gender, and elevated serum sodium and urine volumes in perioperative period were associated with DDAVP requirement. PMID- 16362178 TI - Intra-operative 3D ultrasound in neurosurgery. AB - In recent years there has been a considerable improvement in the quality of ultrasound (US) imaging. The integration of 3D US with neuronavigation technology has created an efficient and inexpensive tool for intra-operative imaging in neurosurgery. In this review we present the technological background and an overview of the wide range of different applications. The technology has so far mostly been applied to improve surgery of tumours in brain tissue, but it has also been found to be useful in other procedures such as operations for cavernous haemangiomas, skull base tumours, syringomyelia, medulla tumours, aneurysms, AVMs and endoscopy guidance. PMID- 16362180 TI - Giant intracranial aneurysm of the anterior communicating artery treated by direct surgery using A3-A3 side-to-side anastomosis and A3-RA graft-STA anastomosis. AB - We describe a giant aneurysm of the anterior communicating artery (ACoA) which was treated with a STA-RA graft-A3 bonnet bypass and A3-A3 side-to-side anastomosis. A giant and partially thrombosed ACoA aneurysm was partially coated 3 years before his current presentation, its gradual increase producing visual field disturbances. An A3-A3 side-to-side anastomosis and STA-RA graft-A3 bonnet bypass were performed. The aneurysm was dissected, and the thrombus removed under transient parent-artery occlusion. The aneurysmal neck was successfully clipped without encountering ischemic changes. This strategy may be useful for treating giant or thrombosed aneurysms in the region of the ACoA. PMID- 16362181 TI - All roads lead to disconnection?--Traumatic axonal injury revisited. AB - Traumatic brain injury (TBI) evokes widespread/diffuse axonal injury (TAI) significantly contributing to its morbidity and mortality. While classic theories suggest that traumatically injured axons are mechanically torn at the moment of injury, studies in the last two decades have not supported this premise in the majority of injured axons. Rather, current thought considers TAI a progressive process evoked by the tensile forces of injury, gradually evolving from focal axonal alteration to ultimate disconnection. Recent observations have demonstrated that traumatically induced focal axolemmal permeability leads to local influx of Ca2+ with the subsequent activation of the cysteine proteases, calpain and caspase, that then play a pivotal role in the ensuing pathogenesis of TAI via proteolytic digestion of brain spectrin, a major constituent of the subaxolemmal cytoskeletal network, the "membrane skeleton". In this pathological progression this local Ca2+ overloading with the activation of calpains also initiates mitochondrial injury that results in the release of cytochrome-c, with the activation of caspase. Both the activated calpain and caspases then participate in the degradation of the local axonal cytoskeleton causing local axonal failure and disconnection. In this review, we summarize contemporary thought on the pathogenesis of TAI, while discussing the potential diversity of pathological processes observed within various injured fiber types. The anterograde and retrograde consequences of TAI are also considered together with a discussion of various experimental therapeutic approaches capable of attenuating TAI. PMID- 16362183 TI - Positron emission tomography in food sciences. PMID- 16362182 TI - Autoreactive antibodies against neurons and basal lamina found in serum following experimental brain contusion in rats. AB - BACKGROUND: Brain trauma is a risk factor for delayed CNS degeneration which may be attenuated by anti-inflammatory treatment. CNS injuries may cause anti-brain reactivity. This study was undertaken to analyze the pattern of delayed post traumatic anti-brain immunity in experimental brain contusion. METHOD: Adult Sprague-Dawley and Lewis rats were subjected to experimental brain contusions. For B-cell investigations, serum was obtained from contused, control and naive rats, and used for immunohistochemistry on slices of rat brains to first detect autoreactive IgG and IgM antibodies in rat serum. Secondly, anti-rat IgG and IgM antibodies were used to search for auto-antibodies already bound to the brain tissue. Double staining with rat-serum and NeuN or anti-GFAP antibody was used to detect anti-neuronal and anti-astrocytic antibodies, respectively. For T-cell reactivity, cells from brains and cervical lymph nodes of rats were used in FACS analysis and elispot with MBP and MOG stimulation. FINDINGS: Anti-vascular basal lamina IgG antibodies were detected at three months in 6/8 rats, following experimental contusion. Anti-neuronal IgG antibodies were detected 2 weeks after experimental contusion and sham surgery, while naive controls were negative. Individual rats showed a prolonged response, or an anti-astrocytic staining. Tissue bound anti-self IgG or IgM was not detected in the brain tissue. Anti-MBP or anti-MOG T-cell responses were not detectable. CONCLUSIONS: Experimental brain trauma and to some degree even sham surgery lead to an individually variable pattern of specific anti-brain reactive B-cells, while a T-cell response did not seem to be a consequence of moderate experimental contusion. The mere presence of anti brain-antibodies may be epiphenomenal, but could also be pathogenic for delayed degeneration. It is reasonable to regard the presence of an actual anti brain reactivity as a potential threat to brain tissue integrity. PMID- 16362186 TI - Feeling and seeing headaches. AB - The aim is to deepen our understanding of headache by three approaches. First, by trying to feel patients' total experience by eliciting their symptoms in detail, and from their reactions to these experiences. Second, by trying to remember one's own experience of headache, and observing a few patients during different headache types. Third, by attempting to see the different mechanisms of headaches by their sites of origin and their pathophysiology. Migraine, tension-type and cluster headache are the three headaches examined by these approaches. Migraine seems to arise from disturbances of the brain's cortex followed by meningeal pain -hence is intracranial in origin. Tension-type headache seems to arise from extracranial muscles, although the pain derives from the fascia or tendons of those muscles; common sites are the masticatory apparatus and the neck--hence extracranial. Cluster headache remains a mystery although vasodilatation provokes, and vasoconstriction stops, attacks--hence vasomotor control is therapeutically valuable. It is concluded that we need more adventurous ideas to deepen our understanding of these and other headaches. PMID- 16362185 TI - Chronic cluster headache: a review. AB - Cluster headache (CH) is a rare but severe headache disorder characterised by repeated unilateral head pain attacks accompanied by ipsilateral autonomic features. In episodic CH, there are periods of headache attacks with pain-free intervals of weeks, months or years in between. A minority of patients have the chronic form, without pain-free intervals between the headache attacks. Chronic CH can occur as primary or secondary chronic CH; the rarest form is episodic CH arising from chronic CH. In this article, we give a review of the chronic forms of CH and focus on demographics, clinical manifestations, social habits, predictive factors, head injury, genetics, neuroimaging and therapy. It is remarkable that little is known about risk factors that make CH chronic. PMID- 16362187 TI - Autonomic features in cluster headache. Exploratory factor analysis. AB - The objective is to identify the pathogenesis of each autonomic manifestation in cluster headache (CH). Through a deductive statistics method (factor analysis) we analysed the type of autonomic symptoms reported by 157 CH patients. Three principal components were identified in the analysis: parasympathetic activation (lacrimation, conjunctival injection and rhinorrhoea), sympathetic defect (miosis and ptosis) and parasympathetic mediated effect (nasal congestion, eyelid oedema and forehead sweating). This work suggests that there are three different mechanisms underlying autonomic manifestations in CH. PMID- 16362188 TI - The present role of percutaneous cervical cordotomy for the treatment of cancer pain. AB - The results obtained by percutaneous cervical cordotomy (PCC) were analysed in 43 terminally ill cancer patients treated in our institution from 1998 to 2001. We wished to determine whether there is still a place for PCC in the actual clinical situation with its wide choice of pain therapies. All patients had severe unilateral pain due to cancer, resistant to opioids and co-analgesics. Following PCC, mean pain intensity was reduced from Numeric Rating Scale (NRS) 7.2 to 1.1. At the end of life, pain had increased to NRS 2.9. Initially following PCC a good result (NRS<3) was obtained in 95% of patients. At the end of life, a good result was still present in 69% of patients. Mean duration of survival after the intervention was 118 days (2-1460). In general, complications were mild and mostly subsided within 3-4 days. There was one case of partial paresis of the ipsilateral leg. PCC remains a valuable treatment in patients with treatment resistant cancer pain and still deserves a place in the treatment of terminal cancer patients with severe unilateral neuropathic or incidence pain. PMID- 16362189 TI - Psychosocial aspects of chronic daily headache. AB - The objective was to investigate possible psychosocial factors in chronic daily headache (HA) by comparing those with chronic daily HA to matched patients with chronic episodic HA and to matched non-HA controls. Although there is some research on psychosocial factors in chronic daily HA, it is conflicting and none to date has compared such patients to both an episodic HA control and a non-HA control. Nineteen patients with chronic daily HA (less than 2% of 4-times-per-day HA ratings were zero) were compared to 19 HA patients matched on age, gender and nominal IHS diagnoses, and to 16 similarly matched non-HA controls on measures of psychological distress (MMPI, BDI, STAI), measures of life stress (major past events, hassles, prospective daily stress) and quality of life (SF-36). Those with chronic daily HA were significantly more distressed and had significantly poorer function on most measures relative to non-HA controls. Although there were many arithmetic trends for chronic daily HA to be more distressed and to function less well than those with episodic HA, only on the depression and social introversion scales of the MMPI and the overall vitality rating of the SF-36 were the differences significant. All three groups had comparable levels of life stress regardless of how it was measured. Those with chronic daily HA have greater levels of psychological distress and poorer quality of life than those with episodic HA or non-HA controls, despite comparable levels of life stress. PMID- 16362190 TI - The patients' perceptions of migraine and chronic daily headache: a qualitative study. AB - This study aimed to gain insight into the management of migraine and chronic daily headache (CDH) from the patients' perspective. This article outlines the patients' perceptions of migraine and chronic daily headache. Thirteen semi structured interviews were carried out with patients suffering from IHS migraine. Five patients, due to their headache frequency of more than 15 headache days per month, were classed as CDH patients. The data were transcribed verbatim and analysed in accordance with the grounded theory methodology. The main themes were: headaches, impact and headaches related to health issues. The theme 'headaches' was sub-divided into 'their pain and symptoms', 'differentiating between their headaches' and 'perceptions of headaches as barriers and facilitators to management'. The patients' perceptions of migraine and CDH were sometimes conflicting and influenced the patients' management behaviours. The qualitative methodology may help to inform doctors, other healthcare professionals and headache researchers about the patients' perspective and possibly develop future headache research, care and education. PMID- 16362191 TI - A non-traumatic interhemispheric subdural haematoma: presented with headache as the sole complaint. AB - Due to their localisations and symptoms, interhemispheric subdural haematomas (ISH) compose a distinct category. Altered level of consciousness and hemiparesis are the most frequent symptoms. We report a case of ISH who presented with headache as the sole complaint. Left cerebellar haematoma and ISH were found in cranial MRI and cranial computed tomography Cranial MR angiogram was normal. Haemogram and coagulation parameters were within normal limits. ISH should be considered among the diagnostic possibilities in elderly patients who present with headache as the sole symptom without other clinical features such as meningeal irritation signs, focal neurological symptoms and alteration of consciousness. Cranial imaging studies should be done in such cases. PMID- 16362192 TI - Observations of the "carry-over effect" following successful termination of chronic migraine in the adolescent with short-term dihydroergotamine, dexamethasone and hydroxyzine: a pilot study. AB - Chronic migraine management almost always requires daily oral preventative medication with potential adverse effects. Daily oral preventative therapy may also not be effective in terminating chronic migraine. Chronic central sensitisation caused by repetitive migraine attacks in a young person may lower the threshold for future migraine episodes leading to an intractable and debilitating disease course. The objective was to determine if short-term parenteral dihydroergotamine, dexamethasone and hydroxyzine can terminate chronic migraine and be followed by a continuous respite or conversion to a more benign episodic form without the need for daily oral preventative medication ("carry over effect"). We treated ten patients, seven adolescents and three adults, with parenteral dihydroergotamine, dexamethasone and hydroxyzine given once a week for a maximum of three weeks. No oral preventative daily medication was administered. The setting was a private practice. Chronic migraine was terminated in all 7 adolescents. Their post-treatment course was converted to a more benign episodic migraine course and no adolescent required daily oral migraine preventative therapy for significantly long carry-over post-treatment observational periods. None of the three adult chronic migraine cases could be terminated satisfactorily as they all required daily oral preventative therapy. In the adolescent group only, this strategy terminated chronic migraine and resulted in a significant carry-over effect that appeared to favourably modify the long-term course without the need for daily pharmacological, potentially toxic, preventive therapy. Although this is a very small study, which requires confirmation by a larger controlled study, our data suggest a significant carryover effect in the young migraineur by administering short-term parenteral dihydroergotamine, dexamethasone and hydroxyzine. PMID- 16362199 TI - Weak segregation theory and non-conventional morphologies in the ternary ABC triblock copolymers. AB - The statistical theory of microphase separation in the ternary ABC triblock copolymers is presented and the corresponding phase diagrams are built both for the linear and miktoarm copolymers. For this purpose the Leibler weak segregation theory in molten diblock copolymers is generalized to multi-component monodisperse block copolymers with due regard for the 2nd shell harmonics contributions defined in the paper. The Hildebrand approximation for the chi parameters is used. The physical meaning of this and alternative choices for the chi-parameters is discussed. The symmetric A(f)B(1-2f)C(f) copolymers with the middle block non-selective with respect to the side ones are shown to undergo the continuous ODT not only into the lamellar phase but also, instead, into various non-conventional cubic phases (depending on the middle block composition it could be the simple cubic, face-centered cubic or non-centrosymmetric phase, which reveals the symmetry of I4(1) 32 space group No. 214 first predicted to appear in molten block copolymers). For asymmetric linear ABC copolymers a region of compositions is found where the weakly segregated gyroid (double gyroid) phase exists between the planar hexagonal and lamellar or one of the non-conventional cubic phases up to the very critical point. In contrast, the miktoarm (star) ABC block copolymers with one of its arm non-selective with respect to the two others are shown to reveal a pronounced tendency towards strong segregation, which is preceded by increase of stability of the conventional BCC phase and a peculiar weakly segregated BCC phase (BCC(3)), where the dominant harmonics belong to the 3rd coordination sphere of the reciprocal lattice. The validity region of the developed theory is discussed and outlined in the composition triangles both for linear and miktoarm copolymers. We present also the list of the 2nd shell harmonics (SAXS reflections) allowed and prohibited in some of the non conventional morphologies due to the weak segregation considerations and comparison of our results with the preceding SCFT treatment of the ABC copolymers by Matsen. PMID- 16362195 TI - A polyconvex anisotropic strain-energy function for soft collagenous tissues. AB - Polyconvexity of a strain-energy function is a very important mathematical condition, especially in the context of a boundary-value problem. In the present paper, we propose an exponential polyconvex anisotropic strain-energy function. It is given by a series with an arbitrary number of terms and associated material constants. Each term of this series a priori satisfies the condition of the energy- and stress-free natural state so that no additional restrictions have to be imposed. Due to the exponential form, the proposed hyperelastic model is suitable for soft biological tissues. Thus, a good agreement with experimental data on different types of tissues is achieved. PMID- 16362200 TI - Stability of cellulose lyotropic liquid crystal emulsions. AB - We studied a new kind of W/O emulsions based on a lyotropic liquid crystal as the aqueous droplet phase. The cholesteric phase, a solution hydroxypropyl cellulose in water was dispersed in the continuous oil matrix, paraffin oil or heptane. We made a specific choice of surfactant in order to impose director anchoring conditions at the oil-water interface and orient the liquid crystal inside the droplet. The strong anchoring conditions resulted in a topological defect inside the droplets of size above the critical value R(*). The defect elastic energy creates a barrier against droplet coalescence, the effect of topological size selection. We have studied the orientation of the director inside the droplets and their size distribution. PMID- 16362201 TI - Reentrant filling transitions in Lennard-Jones fluids confined in nanoscopic slit like pores. AB - The reentrant first-order pore filling transition in a Lennard-Jones (LJ) fluid confined between the parallel (100) planes of the face centered cubic (fcc) crystal is studied by means of Monte Carlo simulations in the canonical as well as grand canonical ensembles. Slit-like pores of the width allowing for the formation of only three adsorbate layers are used. It is demonstrated that the reentrant first-order pore filling transition, associated with the condensation of the middle layer, appears only when the height of the potential barrier between adjacent sites, V(D), fulfills a condition V(D) in [V(D,min),V(D,max)]. The lower limit of V(D) (V(D,min)) is primarily determined by the stability of commensurate monolayers formed at both pore walls during the first step of adsorbate condensation, while the upper limit of V(D) (V(D,max)) depends on the stability of the commensurate three-layer structure. It is also shown that both the misfit between the size of adsorbed atom and the surface lattice as well as the pore width have a great influence on the phase behavior of confined fluids. Moreover, the effects of the phase shift between the confining lattices on the reentrant first-order filling transition are discussed. PMID- 16362203 TI - Fabrication of polymeric scaffolds with a controlled distribution of pores. AB - The design of tissue engineering scaffolds must take into account many factors including successful vascularisation and the growth of cells. Research has looked at refining scaffold architecture to promote more directed growth of tissues through well-defined anisotropy in the pore structure. In many cases it is also desirable to incorporate therapeutic ingredients, such as growth factors, into the scaffold so that their release occurs as the scaffold degrades. Therefore, scaffold fabrication techniques must be found to precisely control, not only the overall porosity of scaffolds, but also the pore size, shape and spatial distribution. This work describes the use of a regularly shaped porogen, sugar spheres, to manufacture polymeric scaffolds. Results show that pre-assembling the spheres created scaffolds with a constant porosity of 60%, but with varying pores sizes from 200-800 microm, leading to a variation in the surface area and likely degradation rate of the scaffolds. Employing different polymer impregnation techniques tailored the number of pores present with a diameter of less than 100 microm to suit different functions, and altering the packing structure of the sugar spheres created scaffolds with novel layered porosity. Replacing sugar spheres with sugar strands formed scaffolds with pores aligned in one direction. PMID- 16362204 TI - Chitosan particles agglomerated scaffolds for cartilage and osteochondral tissue engineering approaches with adipose tissue derived stem cells. AB - It is well accepted that natural tissue regeneration is unlikely to occur if the cells are not supplied with an extracellular matrix (ECM) substitute. With this goal, several different methodologies have been used to produce a variety of 3D scaffolds as artificial ECM substitutes suitable for bone and cartilage tissue engineering. Furthermore, osteochondral tissue engineering presents new challenges since the combination of scaffolding and co-culture requirements from both bone and cartilage applications is required in order to achieve a successful osteochondral construct. In this paper, an innovative processing route based on a chitosan particles aggregation methodology for the production of cartilage and osteochondral tissue engineering scaffolds is reported. An extensive characterization is presented including a morphological evaluation using Micro Computed Tomography (microCT) and 3D virtual models built with an image processing software. Mechanical and water uptake characterizations were also carried out, evidencing the potential of the developed scaffolds for the proposed applications. Cytotoxicity tests show that the developed chitosan particles agglomerated scaffolds do not exert toxic effects on cells. Furthermore, osteochondral bilayered scaffolds could also be developed. Preliminary seeding of mesenchymal stem cells isolated from human adipose tissue was performed aiming at developing solutions for chondrogenic and osteogenic differentiation for osteochondral tissue engineering applications. PMID- 16362205 TI - Polyurethanes as potential substrates for sub-retinal retinal pigment epithelial cell transplantation. AB - Transplantation of cultured retinal pigment epithelial (RPE) cells under the failing macular is a potential treatment for age related macular degeneration. An important step in the development of this procedure is the identification of a suitable membrane on which to grow and transplant the cells. This paper evaluates the potential of using polyurethanes in this application since they possess several of the required properties, such as, flexibility, robustness, biostability and good biocompatiblilty although their hydrophobicity can limit cell adhesion. Three commercially available polyether urethanes (Pellethane, Tecoflex and Zytar) were evaluated in terms of their wettability using dynamic contact angle analysis and their ability to support a monolayer of functioning RPE cells (ARPE-19) . Furthermore Pellethane and Tecoflex were treated with a simple air plasma treatment and analysed as above. In the "as received condition" only a few RPE cells attached to the Pellethane and Tecoflex and remained clumped. RPE cells grew to confluence on the Zytar substrate by 7 days without further surface modification. Air gas plasma treatment of both Pellethane and Tecoflex increased the wettability of the surfaces and this resulted in the growth of a monolayer of well-spread RPE cells on both materials. Morphologically these cells grew with a normal 'cobblestone' phenotype. These results demonstrate the potential of these polyurethanes for this application. PMID- 16362206 TI - Microspheres leaching for scaffold porosity control. AB - Scaffold morphology plays a key role in the development of tissue engineering constructs. The control of pore size, shape and interconnection is needed to achieve adequate nutrient transport and cell ingrowth. Several techniques are available for scaffold manufacturing, but none allows easy control of morphology and is, at the same time, applicable to a wide variety of materials. To investigate the possibility of processing a wide range polymers by solvent casting/particulate leaching with accurate control of scaffold morphology, three different porogens (gelatin microspheres, paraffin microspheres and sodium chloride crystals) were used to fabricate scaffolds from commonly employed biodegradable polymers. The outcome of processing was evaluated in terms of scaffold morphology and structure/properties relationships. Highly porous scaffolds were obtained with all porogens and well defined spherical pores resulted from microspheres leaching. Furthermore, scaffolds with spherical pores showed better mechanical performance and lower flow resistance. Cytocompatibility tests performed showed no evidence of processing residuals released from the scaffolds. Solvent casting/microspheres leaching, particularly gelatin microspheres leaching, can be used to process a large number of polymers and enables to tailor scaffold pore size, shape and interconnection, thus providing a powerful tool for material selection and optimization of scaffold morphology. PMID- 16362207 TI - Nano- and micro-fiber combined scaffolds: a new architecture for bone tissue engineering. AB - One possible interesting way of designing a scaffold for bone tissue engineering is to base it on trying to mimic the biophysical structure of natural extracellular matrix (ECM). This work was developed in order to produce scaffolds for supporting bone cells. Nano and micro fiber combined scaffolds were originally produced from starch based biomaterials by means of a fiber bonding and a electrospinning, two step methodology. The cell culture studies with SaOs-2 human osteoblast-like cell line and rat bone marrow stromal cells demonstrated that presence of nanofibers influenced cell shape and cytoskeletal organization of the cells on the nano/micro combined scaffolds. Moreover, cell viability and Alkaline Phosphatase (ALP) activity for both cell types was found to be higher in nano/micro combined scaffolds than in control scaffolds based on fiber meshes without nanofibers. Consequently, the developed structures are believed have a great potential on the 3D organization and guidance of cells that is provided for engineering of 3-dimensional bone tissues. PMID- 16362208 TI - Microsphere of apatite-gelatin nanocomposite as bone regenerative filler. AB - In this study, we developed novel microspheres comprised of an apatite-gelatin nanocomposite. This microsphere formulation is considered to be useful as a bone regenerative filler, either directly or combined with polymeric matrices. Using the water-in-oil emulsion technique, the apatite-gelatin viscous nanocomposite solution was successfully formulated into microspheres with an average diameter of approximately 110 microm. The microspheres were composed of apatite nanocrystallines precipitated within the gelatin matrix, revealing a typical nanocomposite internal structure. This nanocomposite structure contrasted markedly with that of the conventional composite microspheres which were obtained by directly mixing gelatin with apatite powder. Initial cellular assays showed that the microspheres maintained the adhesion and proliferation of the osteoblastic cells, suggesting the usefulness of the apatite-gelatin nanocomposite microspheres in the bone regeneration field. PMID- 16362210 TI - Hydroxyapatite scaffolds for bone tissue engineering made by 3D printing. AB - Nowadays, there is a significant need for synthetic bone replacement materials used in bone tissue engineering (BTE). Rapid prototyping and especially 3D printing is a suitable technique to create custom implants based on medical data sets. 3D printing allows to fabricate scaffolds based on Hydroxyapatite with complex internal structures and high resolution. To determine the in vitro behaviour of cells cultivated on the scaffolds, we designed a special test-part. MC3T3-E1 cells were seeded on the scaffolds and cultivated under static and dynamic setups. Histological evaluation was carried out to characterise the cell ingrowth. In summary, the dynamic cultivation method lead to a stronger population compared to the static cultivation method. The cells proliferated deep into the structure forming close contact to Hydroxyapatite granules. PMID- 16362209 TI - Biomimetic mineral-organic composite scaffolds with controlled internal architecture. AB - Bone and cartilage generation by three-dimensional scaffolds is one of the promising techniques in tissue engineering. One approach is to generate histologically and functionally normal tissue by delivering healthy cells in biocompatible scaffolds. These scaffolds provide the necessary support for cells to proliferate and maintain their differentiated function, and their architecture defines the ultimate shape. Rapid prototyping (RP) is a technology by which a complex 3-dimensional (3D) structure can be produced indirectly from computer aided design (CAD). The present study aims at developing a 3D organic-inorganic composite scaffold with defined internal architecture by a RP method utilizing a 3D printer to produce wax molds. The composite scaffolds consisting of chitosan and hydroxyapatite were prepared using soluble wax molds. The behaviour and response of MC3T3-E1 pre-osteoblast cells on the scaffolds was studied. During a culture period of two and three weeks, cell proliferation and in-growth were observed by phase contrast light microscopy, histological staining and electron microscopy. The Giemsa and Gomori staining of the cells cultured on scaffolds showed that the cells proliferated not only on the surface, but also filled the micro pores of the scaffolds and produced extracellular matrix within the pores. The electron micrographs showed that the cells covering the surface of the struts were flattened and grew from the periphery into the middle region of the pores. PMID- 16362211 TI - Surface characterization of completely degradable composite scaffolds. AB - The goal of this study was to characterise the surface properties of completely degradable composite, polylactic acid and calcium phosphate glass, scaffolds. The composite scaffolds are made by solvent casting or phase-separation, using chloroform and dioxane as a solvent respectively. The surface properties were measured on composite films which were made using the same procedure as for the three-dimensional (3D) scaffolds without the pore-creating step. The surface morphology, roughness, wettability and protein adsorption capacity of the films was measured before and after sterilisation with ethylene oxide. The results reveal the influence of solvent type, glass weight content and sterilisation on the wettability, surface energy and protein adsorption capacity of the materials. The addition of glass particles increase the hydrophylicity, roughness and protein adsorption capacity of the surface. This effect, however, depends on the extent of the coating of the glass particles by the polymer film, which is much higher for dioxane films than for chloroform films. This information can be used to interpret and understand the biological behaviour of the 3D scaffolds made of this composite materials. PMID- 16362212 TI - Soluble phosphate glass fibres for repair of bone-ligament interface. AB - Phosphate-based fibres of the generic composition (CaO)0.46-(Na2O)n-(Fe2O3)y (P2O5)0.50 have been evaluated, in vitro, as three dimensional scaffolds for tissue engineering of the hard-soft tissue interface by assessing the fibre solubility and growth and functional gene expression of human cells. Primary human osteoblasts and fibroblasts were seeded onto scaffolds and maintained in culture for up to 21 days. Fluorescent immunolabeling revealed the spread cell morphology and significant proliferation pattern on these fibres, particularly on the 3 mol% Fe2O3-containing formulation. Real-time quantitative Polymerase Chain Reaction (rtQ-PCR) analysis of gene expression using TaqMan Probes was preformed and it has been established that committed cell differentiation was maintained by both cell types, and was strongly related to the 3 mol% Fe2O3 glass composition. These novel, readily manufactured, soluble glass fibres offer a biocompatible and biochemically favourable alternative in the search for suitable degradable materials used in Tissue Engineering. PMID- 16362213 TI - Novel deposition of nano-sized silicon substituted hydroxyapatite by electrostatic spraying. AB - Suspensions containing nano-sized silicon substituted hydroxyaptite (nSiHA) particles were produced and processed for electrostatic spray deposition. No secondary phases were detected by X-ray diffraction, which indicated that the nSiHA was phase pure. Electrostatic spraying of nSiHA in cone-jet mode was achieved at flow rate of 10(-9) m3s(-1) with an applied voltage between the needle and the ring-shaped ground electrode set at 6 to 8 kV. Micrometer- and submicrometer-scaled islands of nSiHA have been deposited on glass and titanium substrates. The surface roughness of such nHA and nSiHA islands was in the range 60 to 80 nm, as measured from atomic force microscopy in tapping mode. The growth of primary human osteoblast (HOB) cells on the nSiHA deposited substrates increased with time during the 4 days of culture, and the increase was related with the Si content in substituted HA, indicating that nSiHA was able to promote and support the growth of HOB cells. Scanning electron microscopy (SEM) revealed that extracellular matrix (ECM) produced by the HOB cells on these nSiHA deposits was well organized. In addition, the presence of Ca and P containing nodules in the ECM were also confirmed by Energy Dispersive X-ray (EDX) analysis, indicating early signs of calcification fronts. The results showed that nSiHA produced by electrostatic spray deposition was able to promote the attachment and the growth of HOB cells. Therefore, electrostatic spray deposition offers great potential for the creation of bioactive surfaces to provide improved interfacial bonding with host tissues. PMID- 16362214 TI - Crystal imperfection studies of pure and silicon substituted hydroxyapatite using Raman and XRD. AB - Hydroxyapatite (HA) is important in biomedical applications because of its chemical similarity to the mineral content of bone and its consequent bioactivity. Silicon substitution into the hydroxyapatite crystal lattice was found to enhance its bioactivity both in vitro and in vivo [1, 2]. However, the mechanism for the enhancement is still not well understood. In this paper, the crystal imperfections introduced by silicon substitution were studied using XRD and Raman spectroscopy. It was found that silicon substitution did not introduce microstrain, but deceased the crystal size in the hk0 direction. Three new vibration modes and peak broadening were observed in Raman spectra following silicon incorporation. The imperfections introduced by silicon substitution may play a role in enhancing bioactivity. A phenomenological relationship between the width of the PO4 v1 peak and crystal size was established. PMID- 16362216 TI - Porous Ti6Al4V scaffolds directly fabricated by 3D fibre deposition technique: effect of nozzle diameter. AB - 3D porous Ti6Al4V scaffolds were successfully directly fabricated by a rapid prototyping technology: 3D fibre deposition. In this study, the rheological properties of Ti6Al4V slurry was studied and the flow rate was analyzed at various pressures and nozzle diameters. Scaffolds with different fibre diameter and porosity were fabricated. ESEM observation and mechanical tests were performed on the obtained porous Ti6Al4V scaffolds with regard to the porous structure and mechanical properties. The results show that these scaffolds have 3D interconnected porous structure and a compressive strength which depends on porosity at constant fibre diameters and on the fibre diameter at constant porosity. These Ti6Al4V scaffolds are expected to be constructs for biomedical applications. PMID- 16362215 TI - Investigation of a novel freeze-thaw process for the production of drug delivery hydrogels. AB - Poly(vinyl alcohol) (PVA) is a water-soluble, biocompatible and biodegradable polymer, which has been widely applied in biomedical fields. In this paper, novel physically cross-linked hydrogels composed of PVA and comprising a blend of poly(vinyl alcohol) (PVA) with different concentrations of HCl, NaOH and NaCl are prepared by a freezing/thawing treatment of aqueous solutions. The structure and complexation of the electrolytes were studied by Fourier transform infrared (FTIR) spectroscopy. The mechanical properties were investigated using rheometery and the thermal transitions of the hydrogels were examined by modulated differential scanning calorimetry (MDSC). Freeze/thawed PVA gels containing NaOH showed overall enhanced swelling with increased mechanical strength over traditional gels prepared by chemical or irradiative crosslinking techniques. These novel physically cross-linked hydrogels show promise for a variety of biomedical and drug delivery applications. PMID- 16362217 TI - Titanium--hydroxyapatite porous structures for endosseous applications. AB - Materials for uncemented endosseous implants have to assure an as short as possible osseointegration time. Thus, a material with both surface bioactivity and a porous outer structure can become a preferred choice for this type of applications. This paper presents a class of titanium-base PM composites, reinforced with particulate hydroxyapatite. Raw materials were titanium powder, obtained through hydriding--milling--dehydriding, with the grain size of 63-100 microm, and sol-gel hydroxyapatite (HA) powder, produced by the reaction between Ca(NO3)2 x 4H2O and (NH4)2HPO4. Blends with 5 to 50% HA were prepared and pressed in a rigid die, producing single composition or gradual composition samples. The applied pressure was of 400, 500 or 600 MPa. Sintering was performed in vacuum, at 1160 ( composite function)C. All samples, although well sintered, displayed swelling during sintering, due to diffusion into the matrix. The increase in volume is more severe for higher amounts of HA in the green compacts and for higher applied compaction pressure. Compacts with a gradual increase of the HA content are recommended from the functional and mechanical point of view, but the increase should be slow, not to produce interlayer cracks. The outer surface shows interconnected pores, suitable for the ingrowth of vital new bone. PMID- 16362219 TI - Effects of TiO2, ZrO2 and Al2O3 dopants on the compressive strength of tricalcium phosphate. AB - Tricalcium phosphate (TCP) powders synthesised using the Ca(NO3)2 and Ca(OH)2 routes were doped with TiO2, ZrO2 and Al2O3 in order to increase their compressive strength. An ultimate compressive strength (UCS) of 255 +/- 6 MPa was achieved for approximately 10 vol% TiO2 doping compared to 30 +/- 3 MPa for an un doped control processed and tested in the same manner. Higher levels of TiO2 doping resulted in smaller increases in UCS with 30 and 50 vol% achieving 213 +/- 9 and 178 +/- 15 MPa, respectively. Very small amounts of Al2O3 doping (< 0.5 vol%) also resulted in a stronger materials. However, under the processing conditions employed, higher levels of Al2O3 and ZrO2 doping resulted in no beneficial effect on the UCS. Polyvinyl alcohol (PVA) was used as binding agent to facilitate processing. As expected, higher levels of PVA were associated with smaller increases in UCS. Powders synthesised using the Ca(OH)2 route had smaller particle size and resulted in larger increases in UCS compared to the Ca(NO3)2 synthesised powders. Although some powders contained alpha and beta-TCP phases, no other calcium phosphate, CaO, CaTiO3 or CaZrO3 phases were detected. In conclusion, a significant increase in the UCS of TCP was achieved by doping with approximately 10 vol% TiO2 which is expected to have little or no effect on the bioactivity or bioresorbability of the material. PMID- 16362218 TI - TiO2 type influences fibronectin adsorption. AB - Human fibronectin (FN) plays a key role in the biointegration of implants as the success depends on adsorption of proteins like FN [1]. Indeed FN can be an intermediary between the biomaterial surface and cells. The adsorption of human fibronectin (FN) on commercially pure titanium with a titanium oxide layer formed in a H2O2 solution (TiO2 cp) and TiO2 sputtered on Si (TiO2 sp) was studied. Adsorption isotherms and the work of adhesion were assessed by wettability studies, X-ray photoelectron spectroscopy (XPS), and by radiolabelling of FN with 125I, (125)I-FN. Exchangeability of bound FN by free FN, was also evaluated by the radiolabelling technique. Contact angle determinations have shown that FN displays higher affinity for the TiO2 cp surface than for the TiO2 sp. As expected from the surface free energy values, the work of adhesion of FN is higher for the TiO2 cp substrate, the more hydrophilic one, and lower for the TiO2 sp substrate, the more hydrophobic one. The adsorption isotherms were evaluated by two different techniques: radiolabelling of FN (125I-FN) and XPS. TiO2 cp adsorbs more FN than the TiO2 sp surfaces as shown by the radiolabelling data. FN molecules are also more strongly attached to the former surface as indicated by the work of adhesion and by the exchangeability studies. Results using 125I-FN also suggests that FN adsorbs as a multilayer for FN concentrations in solution higher than 100 microg/mL. PMID- 16362220 TI - Cement from nanocrystalline hydroxyapatite: effect of calcium phosphate ratio. AB - Nanocrystalline hydroxyapatite (nHA) can be mixed with phosphoric acid to form a brushite cement; a degradable inorganic bone filling material. nHA was precipitated from reactants of calcium to phosphate (Ca/P) ratio 0.8 to 2.0 and mixed with phosphoric acid, which resulted in the formation of a brushite cement. Cement was also formed by mixing microcrystalline calcium phosphates, beta tricalcium phosphate, hydroxyapatite and tetracalcium phosphate with phosphoric acid solution. Cement produced with nHA was stronger in compression than that formed with crystalline calcium phosphate phases. Setting time, strength and composition of cement produced with nHA was dependant on both the Ca/P ratio of nHA and the concentration of phosphoric acid in cement slurry. Increasing phosphoric acid concentration increased compressive strength whilst reducing the initial setting time of cement. Reducing the Ca/P ratio of nHA precipitation reactants retarded the setting and increased the extent of reaction of cements. This finding was unexpected and suggests that Ca/P ratio may strongly affect dissolution behaviour and this parameter is more important than stoichiometry in determining extent of reaction in this system. This study demonstrated that the wide variation in stoichiometry that may be attained in nanocrystalline apatite may be utilised to change cement performance and setting behaviour. PMID- 16362224 TI - In-vivo short- and long-term evaluation of the interaction material-blood. AB - Through an innovative electron microscopy technique, thrombi and fibrotic tissue taken from 14 explanted vena cava filters were observed. Twelve cases showed the presence of micro-and nano-sized inorganic, non biodegradable nor biocompatible particles which did not belong to the metal the device was made of and which could be the sole cause or, more likely, a pre-existing cause for thrombosis. In two cases, those debris activated immunological reactions typical of a foreign body. The presence of inorganic particles in the blood was never detected before and their effects on human health are hardly known. Their thrombogenicity should be added to the Virchow's Triad as a fourth factor and could be the explanation to many of the cases of pulmonary embolism where no thrombotic focus could be demonstrated. PMID- 16362221 TI - An experimental and theoretical composite model of the human mandible. AB - The purpose is to design and manufacture a composite mandible replicate suitable for testing the influence of prosthetic materials on the stress distribution of bone. Composite mandibles made of a poly(methylmethacrylate) core and a glass reinforced outer shell are manufactured and characterised through mechanical tests assisted by the finite element analysis. The mandible replicate has been conveniently equipped with strain gauges, moreover a video extensometer has also been used in order to measure the arch width change during loading. A close agreement is found between the experimental data and the theoretical predictions. By laterally loading the mandibles the maximum values of stress and strain take place in the premolar-incisal region. By varying technological parameters such as the fiber volume fraction and orientation, it is easy to replicate the behaviour of mandibles having different stiffnesses. The results obtained by laterally loading the composite mandibles through the condyles or through the gonion regions are consistent with literature data relative to the arch width decrease of natural jaws during opening and closing. This novel synthetic system coupled with the Finite Element model constitutes an experimental-theoretical model suitable to investigate the biomechanical effects of oral rehabilitations on mandibular bone. PMID- 16362223 TI - Inflammatory response to a novel series of siloxane-crosslinked polyurethane elastomers having controlled biodegradation. AB - A series of polyurethane polymers was synthesized with increasing proportions of silicone in the form of polydimethylsiloxane (PDMS) utilised as a cross-linking agent, based on an aromatic, non-biostable polyetherurethane (PEtU). Eight formulations ranging from 0-50% PDMS were constructed into porous and non-porous films. These were implanted subcutaneously in rats, both unstrained and 100% strained, for 3 and 6 months. Degradation was determined by FTIR-ATR. Porous films were implanted for 6 and 12 months intramuscularly in both rats and rabbits. These were explanted and examined for inflammatory cell markers by immunohistochemistry. Both low and high percentages of siloxane gave rise to increased degradation, with 20-40% PDMS resulting in the least degradation. Infrared spectral changes correlated well with both visual examination and observation by SEM. Changes to the concentration of siloxane gave rise to differences in the thickness of fibroblastic capsule and infiltration of inflammatory cells in both films & scaffolds. Cellular infiltration was greatest in the films with lower siloxane concentrations. Macrophage activation (MHC-I & MHC-II expression) was least in the higher siloxane variants. It is concluded that by varying the siloxane content in the PEtU matrix we can obtain an acceptable inflammatory response with a relatively short degradation time. PMID- 16362225 TI - Physical and biological characterizations of a novel multiphase anodic spark deposition coating to enhance implant osseointegration. AB - The present study assessed in vitro the short-term cellular response to surface physico-chemical properties of a new, purposed bioactive surface treatment called BioSpark performed on simply machined and on sand-blasted titanium. Material characterisation was carried out using scanning electron microscopy, energy dispersion spectroscopy, laser profilometry, and thin film X-ray diffraction. The in vitro biological study showed a suitable cellular response with adhesion and spreading level comparable for all the tested specimens. The proliferation analysis demonstrated that all the surfaces successfully supported cellular colonisation; in particular, higher cellular proliferation activity was observed on the BioSpark-treated materials, with values higher than machined titanium. The results suggest that the BioSpark treatment represents a smart way to enhance osteoblastic cellular colonisation and thus improve osteointegration processes of machined and sandblasted titanium for orthopaedic and dental implants. PMID- 16362222 TI - Bioceramics composition modulate resorption of human osteoclasts. AB - Biomaterials used in bone regeneration are designed to be gradually resorbed by the osteoclast and replaced by new bone formed through osteoblastic activity. The aim of the present study is to analyze the role of osteoclasts in the resorption process. The attachment of human osteoclasts and the appearance of their resorption lacunae, when cultured on either the resorbable crystalline, calcium orthophosphate materials or on the long-term stable bioceramic material was investigated. The resorbable materials contain Ca10[K,Na](PO4)7 (AW-Si) and Ca2KNa(PO4)2 (GB14, GB9 & D9/25) as their main crystal phases, however they differ in their total solubility. These differences result from small variations in the composition. The long-term stable material consist of about 30% fluorapatite beside calcium zirconium phosphate (Ca5(PO4)3F + CaZr4(PO4)6) and shows a very small solubility. AW-Si has an alkali containing crystalline phase, Ca10[K,Na](PO4). While GB14, GB9 and D9/25 contain the crystalline phase Ca2KNa(PO4)2 with small additions of crystalline and amorphous diphosphates and/or magnesium potassium phosphate (GB14). D9/25 and AW-Si is less soluble compared to GB14, and GB9 among the resorbable materials. Resorbable and long term stable materials vary in their chemical compositions, solubility, and surface morphology. Osteoclasts modified the surface in their attempts to resorb the materials irrespective of the differences in their physical and chemical properties. The depth and morphology of the resorption imprints were different depending on the type of material. These changes in the surface structure created by osteoclasts are likely to affect the way osteoblasts interact with the materials and how bone is subsequently formed. PMID- 16362226 TI - Nitinol Carbofilm coated stents for peripheral applications: study in the porcine model. AB - PURPOSE: Testing the safety and foreign body reaction (FBR) of a nitinol self expandable carbon-coated stent system in the porcine animal model at different follow-up (FU) periods. METHODS: Fifteen minipigs received 30 carbon-coated self expandable nitinol stents in iliac arteries. Explants were carried out at 7 (3 animals), 30 (4 animals), 90 (4 animals) and 180 (4 animals) day FU, for evaluation of acute, sub-acute and chronic biological response to the implanted devices. Histological, immunohistochemical, histomorphometric and scanning electron microscopy (SEM) analyses were performed to assess inflammatory reaction, endothelialization process, neointimal growth and cellular composition. RESULTS: Thirty stents were successfully implanted. No mural thrombi were observed at gross examination or by angiography. Histologically no significant inflammatory reaction was detected: the stents appeared covered by a thin monolayer of endothelial cells even at 7 day FU. The neointima presented homogeneous growth and moderate thickness after 30, 90 and 180 days explants (0.38+/- 0.36 mm, 0.33+/- 0.30 mm, 0.27+/- 0.25 mm respectively). Internal and external elastic laminae were intact in 95% of stented arteries. Histological data validations of vessel endothelialization was obtained with SEM for the seven day follow-up group. CONCLUSIONS: This study showed good remarkable technical performances, minimal FBR and biocompatibility comparable with other available pre-clinical experimentations. PMID- 16362227 TI - Evaluation of the relationship between inducible nitric oxide synthase (iNOS) activity and effects of melatonin in experimental osteoporosis in the rat. AB - Inducible nitric oxide synthase (iNOS) plays a critical role in the pathogenesis of osteoporosis. iNOS generates nitric oxide (NO), a free radical contributing to the imbalance between bone formation and resorption caused by estrogen depletion. Melatonin is the major product of the pineal gland which is known to diminish iNOS expression and NO production significantly. The aim of this study was to determine the distribution of iNOS and the amount of apoptotic cells after melatonin treatment in ovariectomized rats. Since previous studies have shown that constitution of bone formation is primarily sustained in nucleus pulposus and epiphyseal cartilage, experiments were carried out on nucleus pulposus and epiphyseal cartilage; additional quantitation of osteoblasts and osteoclasts were evaluated on vertebral area as well. Vertebral sections of ovariectomized rats were obtained from formalin-fixed and parafin-embedded blocks. iNOS expression and quantitation of apoptotic cells in nucleus pulposus and epiphyseal cartilage were evaluated using indirect immunoperoxidase and TUNEL techniques, respectively. The number of osteoclasts and osteoblasts in trabecular bone was determined using histomorphometry. Ovariectomy increased iNOS expression and the number of apoptotic cells in nucleus pulposus and epiphyseal cartilage, whereas a 4-week treatment with melatonin (10 mg/kg/day) resulted in the reduction of both effects. These data indicate that there is strong influence of melatonin application on expression of iNOS, apoptosis, osteoclast and osteoblast numbers after ovariectomy. In conclusion, melatonin besides its usual use as an antiaging hormone, may also be an effective hormone in treatment of bone changes in estrogen deficiency states. PMID- 16362229 TI - The hamate facet of the lunate: a radiographic study in an Arab population from Bahrain. AB - Based on the presence of a facet for the hamate two types of lunate bone have been described in the wrist; type I is without a facet for the hamate while type II bears a facet on the medial surface for articulation with the proximal pole of the hamate. Compared to type I lunates, type II lunates have been shown to have different kinematics, are more prone to clinically relevant degenerative changes in the hamato-lunate joint and may cause ulnar-sided wrist pain. The incidence of type II lunates seems to vary in different populations. Postero-anterior (PA) view of plain radiographs of the hand was studied in Arab subjects from Bahrain (n=381; 307 males and 74 females) to determine the incidence of type I and type II lunates. The mean age of the subjects was 31.1 years, the range being 18-80 years. In the population studied, only 148 (38.8%) of the 381 wrists showed type II lunate; 121 (31.7%) were found in males and 27 (7.1%) in females. There was no significant difference in the mean age of subjects with type I or type II lunates (P=0.16). There was no correlation between side and gender with the presence or absence of a hamate facet on the lunate. The incidence of type II lunate was much lower than reports in the literature of populations from the Western world, which could be attributed to a genetic variation in the Arab population in Bahrain. These data indicate that osteoarthritis of the hamate or lunate is less likely to be a cause of ulnar-sided wrist pain in Bahraini Arabs. PMID- 16362228 TI - A common hepatic artery passing in front of the portal vein. AB - A variation in liver vascularization was discovered in a 50-year-old man. A single common hepatic artery was found to be responsible for vascularization of the entire liver. This artery was unusual in that it formed the first branch of the superior mesenteric artery, crossing the portal trunk shortly after its origin, and passed in front of the portal vein to reach the hilum of the liver, where it divided into a right and a left branch. This artery was a true common hepatic artery because a gastroduodenal artery emerged from it 2 cm after its origin. A common hepatic artery originating from the mesenteric artery and passing in front of the portal vein has never been described before. The patient had a second anatomical variation: the left gastric artery and the splenic artery arose directly from the aorta, without celiac trunk separation. This observation confirms the importance of carrying out a precise vascular assessment before all types of hepatic or pancreatic surgery, to identify possible variations in the number or trajectory of hepatic arteries. PMID- 16362230 TI - Is there a risk of infertility after inguinal mesh repair? Experimental studies in the pig and the rabbit. AB - The implantation of a non-absorbable polypropylene mesh during hernia repair causes chronic foreign body reaction involving the surrounding tissue. In case of inguinal hernia repair using mesh techniques, the spermatic cord is potentially affected by this chronic inflammatory tissue remodeling. This effect has been investigated using standardized animal models (pig and rabbit). Fifteen adult male pigs underwent transinguinal preperitoneal implantation of a polypropylene mesh. The contralateral side with a Shouldice repair served as control. After 7, 14, 21, 28, and 35 days, three animals were sacrificed. The spermatic cords were resected and analyzed histologically. In a second experiment Lichtenstein repair using the same polypropylene mesh and Shouldice repair on the contralateral side was done in eight chinchilla rabbits. Three animals served as controls. Three months after operation, the analysis included testicular size, testicular temperature, and testicular and spermatic cord perfusion. We added histological evaluation of the foreign body reaction and the spermatogenesis using the Johnsen score. In the pig, we observed a certain foreign body reaction with diffuse infiltrating inflammatory cells after mesh implantation. Venous thrombosis of the spermatic veins occurred in five of 15 cases. One animal presented focal fibrinoid necrosis of the deferent duct wall. The side of Shouldice repair showed only minor postoperative changes. In the rabbit, we also observed a typical foreign body reaction at the interface between mesh and surrounding tissue, which was not detectable after Shouldice repair. The mesh repair led to a decrease of arterial perfusion, testicular temperature, and the rate of seminiferus tubules with regular spermatogenesis classified as Johnsen 10 (Lichtenstein: 48.1%, Shouldice: 63.8%, controls: 65.8%). Testicular volume increased about 10% after each operation. The implantation of a polypropylene mesh in the inguinal region induces major response of the structures of the spermatic cord. This may have an influence also on spermatogenesis. Due to this a strict indication for implantation of a prosthetic mesh during inguinal hernia repair is recommended. PMID- 16362231 TI - A continuous debate on contralateral processus vaginalis: evaluation technique and approach to patency. AB - We present our experience with the transinguinal diagnostic laparoscopy and discuss its efficacy in evaluating the contralateral side in unilateral inguinal hernias. The possible influence of the initial side of hernia, age and gender on recognized bilateralism were also evaluated in two study groups (diagnostic laparoscopy vs clinical diagnosis). In this retrospective study, we evaluated 36 bilateral, 158 left-sided and 303 right-sided consecutive inguinal hernia cases. A total of 211 out of 461 unilateral cases underwent hernia repair and transinguinal laparoscopic evaluation of the contralateral side. Complications and difficulties of the technique, the mean duration of laparoscopy and operative times were additionally analyzed from operation charts. In this study, bilateralism was determined by transinguinal laparoscopy in Group 1 (children with a contralateral patent processus vaginalis) and clinically in Group 2 those who had a metachronous hernia. The patients were also analyzed according to the side of the inguinal hernia, age and gender in both groups. We found an overall positive contralateral patency in 41 cases (19.4%). We failed to perform a successful diagnostic laparoscopy in six cases (2.7%). No anesthetic or surgical complications were noted. Transinguinal laparoscopy did not add any considerable time to the mean operation time. The incidence of bilateralism in Group 1 was higher in girls than boys. Clinically detected bilateralism was significantly higher in 0-6 month age group and contralateral patency detected via laparoscopy was high in all other age groups. We may conclude that transinguinal diagnostic laparoscopy is a feasible technique in children. We advocate its use as a tool with minimal complication risk even in the hands of novice. PMID- 16362232 TI - A linkage study of academic skills defined by the Queensland core skills test. AB - This study used genome-wide linkage analysis to detect Quantitative Trait Loci (QTLs) implicated in variation in general academic achievement as measured by the Queensland Core Skills Test (QCST) (Queensland Studies Authority, 2004). Data from 210 families were analysed. While no empirically derived significant or suggestive peaks for general academic achievement were indicated a peak on chromosome 2 was observed in a region where Posthuma et al. (2005) reported significant linkage for Performance IQ (PIQ) and suggestive linkage for Full Scale IQ (FSIQ), and Luciano et al. (this issue) observed significant linkage for PIQ and word reading. A peak on chromosome 18 was also observed approximately 20 cM removed from a region recently implicated in reading achievement. In addition, on chromosomes 2 and 18 peaks for a number of specific academic skills, two of which were suggestive, coincided with the general academic achievement peaks. The findings suggest that variation in general academic achievement is influenced by genes on chromosome 2 which have broad influence on a variety of cognitive abilities. PMID- 16362233 TI - Maternal health-seeking behavior and associated factors in a rural Nigerian community. AB - OBJECTIVES: To assess maternal health services and health-seeking behavior in a rural community (Ologbo), located in the South-south zone of Nigeria. METHODS: Structured questionnaire was administered to 225 randomly selected mothers (age 15-49 years), and was analyzed using SPSS. Six focus group discussion sessions were also conducted-four for community women and two for health workers. RESULTS: Teenagers constituted 13.3% of the respondents. The average number of children per woman ranged from 2.5 for teenagers to 9.0 for women aged 45-49 years. Eighty percent of respondents knew at least one major medical cause of maternal mortality: the most common causes mentioned were hemorrhage (31.8%) and obstructed labor (17.3%). Private maternity center was the most preferred place for childbirth (37.3%), followed by traditional birth attendants (TBAs) (25.5%). Government facility was preferred by only 15.7%: reasons for the low preference included irregularity of staff at work (31.4%), poor quality of services (24.3%), and high costs (19.2%). Among the 81 women that delivered within a 1-year period, only 9.9% received antenatal care, 6.2% received two doses of tetanus toxoid, while 4.9% attended postnatal clinic. Private midwives and TBAs attended 49.4 and 42.0% of deliveries, respectively. Education was found to be significantly associated with choice of place for delivery (p < 0.05), but no association was found with respect to age and marital status. Only 11.4% of mothers were practicing family planning. CONCLUSIONS: Poor health-seeking behavior is a challenge in rural Nigeria, and interventions are needed to achieve improved maternal health status. PMID- 16362234 TI - State-specific trends in preterm delivery: are rates really declining among non Hispanic African Americans across the United States? AB - OBJECTIVES: This study sought to examine state-specific trends in preterm delivery rates among non-Hispanic African Americans and to assess whether these rates are influenced by misclassification of gestational age. METHODS: The sample population consisted of singleton non-Hispanic White and non-Hispanic African American infants born in 1991 and 2001 to U.S. resident mothers. For both time periods, state-specific and national preterm delivery rates were calculated for all infants, stratified by infant race/ethnicity. Next, birth-weight distributions within strata of gestational age were studied to explore possible misclassifications of gestational age. Lastly, state-specific and national preterm delivery rates among infants who weighed less than 2,500 g were separately computed. RESULTS: National analyses showed that the frequency of preterm delivery increased by 15.8% among non-Hispanic Whites but declined by 10.3% among non-Hispanic African Americans over the same period. For both subgroups, a bimodal distribution of birth weights was apparent among preterm births at 28-31 weeks of gestation. The second peak with its cluster of normal weight infants was more prominent among non-Hispanic African Americans in 1991 than in 2001. After excluding preterm infants who weighed 2,500 g or more, the national trends persisted. State-specific analyses showed that preterm delivery rates increased for both subgroups in 13 states during this period. Of these 13, 6 states had a number of non-Hispanic African-American births classified as preterm that were apparently term births mistakenly assigned short gestational ages. Such misclassification was more frequent in 1991 than in 2001 and inflated 1991 rates. CONCLUSION: There is heterogeneity in state-specific preterm delivery rates. Such differences are often overlooked when aggregate results are presented. PMID- 16362237 TI - The impact of exposure to transphobia on HIV risk behavior in a sample of transgendered women of color in San Francisco. AB - This study examined the relationship between exposure to transphobia--societal discrimination and stigma of individuals who do not conform to traditional notions of gender--and risk for engaging in unprotected receptive anal intercourse (URAI) among 327 transgendered women of color. Overall, 24% of participants had engaged in URAI at least once in the past 30 days. Individuals who self-identified as pre-operative transsexual/transgendered women were significantly more likely than self-identified females to have engaged in URAI. Although exposure to transphobia was not independently related to URAI, an interaction between age and experiencing discrimination was observed. Among transgendered women 18-25 years old, those reporting higher levels of exposure to transphobia had a 3.2 times higher risk for engaging in URAI compared to those reporting lower levels. Findings from this study corroborate the importance of exposure to transphobia on HIV risk, particularly among transgendered young adults. PMID- 16362236 TI - A comparison of birth outcomes among US-born and non-US-born Hispanic Women in North Carolina. AB - OBJECTIVE: To compare birth outcomes between non-US-born and US-born Hispanic women in North Carolina (NC). METHODS: A retrospective comparison of birth outcomes from linked NC birth/death certificate data (1993-1997) for 22,234 Hispanic births by mother's place of birth was conducted. RESULTS: Mexico-born Hispanic women (58%) had significantly fewer medical risks, tobacco or alcohol use during pregnancy; however, they also had significantly less education and prenatal care than US-born Hispanic women (21%). Infant mortality rate, low birth weight, and prematurity were low and did not differ significantly. Lethal anomalies were the primary cause of infant mortality in non-US-born Hispanics versus Sudden Infant Death Syndrome (SIDS) in US-born Hispanics. CONCLUSIONS: Despite increased risk factors among US-born women, we found no difference in Hispanic birth outcomes in NC by mother's place of birth. These data contradict national data and may be related to findings of both positive and negative aspects of acculturation in NC. PMID- 16362235 TI - Spousal abuse against women and its consequences on reproductive health: a study in the urban slums in Bangladesh. AB - OBJECTIVES: Spousal violence is common and results in costly problems both for society and for the reproductive health of women. Despite the recognition that violence may be associated with serious consequences for women's reproductive health, our understanding of the relationship between the two remains limited. In this study, we assessed the association between spousal violence and women's reproductive health. METHODS: Data from an interviewer-administered questionnaire assessing socioeconomic, demographic, behavioral profiles, and spousal violence related information was collected from 496 women. Subjects were chosen from eight randomly selected urban slums from four metropolitan cities. RESULTS: Spousal violence was significantly higher amongst the group of less educated women who had been in several marriages; indicating that the social and behavioral traits of women act as catalysts for spousal violence. Abusive husbands also had been married several times and were more likely to be addicted to alcohol or drugs. This demonstrates that the behavioral traits of husbands were also responsible for spousal violence. Spousal violence injuries adversely affect the health and well-being of women. More than three-quarters of physically violated women suffered injuries as a result of this violence. About 50% of these injuries were minor and about 10% serious. Sexual violence adversely affected women's health; more than 80% of sexually violated women complained of pelvic pain, more than 50% reported reproductive tract infections, and more than 50% reported symptoms of irritable bowel syndrome. Abused women suffered from gynecological problems at the time of pregnancy significantly more than non-abused women (p<0.05) and abused women suffered from reproductive tract infections significantly more than non-abused women (p<0.01). Abusive husbands suffered from sexually transmitted diseases (STD) significantly more than non-abusive husbands (p<0.05). Abused women used contraception significantly less than non-abused women (p<0.01). Logistic regression analysis suggested that spousal violence was the most important contributing factor for reproductive health problems in women. CONCLUSIONS: The findings of this study may enhance our understanding of the impact of spousal violence against women and their reproductive health and therefore highlight the importance of spousal violence prevention measures. Increasing the awareness and understanding of the relationship between violence against women and reproductive health could be achieved if lawyers, researchers, clinicians, practitioners, and government workers from multiple disciplines and agencies worked together. PMID- 16362238 TI - Prospective study of the effectiveness of coping in pediatric patients. AB - Findings about the influence of coping on psychological adjustment in children with different medical conditions are inconsistent and often based on cross sectional data. This prospective study evaluated the effect of various coping strategies on children's post-traumatic stress symptoms and behavioral problems 1 month and 1 year after an accidental injury or the diagnosis of a chronic disease in 161 pediatric patients 6-15 years of age. Only minor positive effects of coping on psychosocial adjustment were found: Religious coping reduced post traumatic stress symptoms. Active coping strategies had negative effects on internalizing and externalizing behavioral problems. Support seeking strategies, distraction, and avoidance had no impact on long-term psychosocial adjustment. PMID- 16362239 TI - Child and family predictors of therapy outcome for children with behavioral and emotional problems. AB - This study investigated the characteristics of 106 children primarily referred for externalizing behavior problems and their families, and assessed the prediction of treatment outcome following a standardized short-term, cognitive behavioral group program. Exploring Together comprised a children's group (anger management, problem-solving and social skills training), a parents' group (parenting skills training, dealing with parents' personal, relationship and family-of-origin issues), and a combined children's and parents' group (to target parent-child interactions). The main predictors of reductions in externalizing and internalizing behaviors at home following treatment were children's pre existing levels of these behavioral and emotional problems (children with higher levels improved most), and positive parent-child interaction. Higher pre-existing levels of behavioral and emotional problems and low levels of attention problems predicted greater improvement in post-treatment school behaviors. Implications of the findings for improving interventions for childhood behavioral and emotional problems are discussed. PMID- 16362245 TI - Sexual orientation and gender: stereotypes and beyond. AB - In this essay, I explore empirical research about the relationship between sexual orientation and gender. Both concepts have multiple meanings, which hinder an unequivocal understanding of the relationship between the two concepts. One of the first studies assumed to establish a relationship between male homosexuality and femininity was conducted by L. M. Terman and C. C. Miles (1936). The study actually gives a much more complicated picture of the relationship. Terman and Miles actually found two groups of homosexual men with, respectively, extremely high masculinity and extremely high femininity scores. Subsequently, various other studies examined the relationship between sexual orientation and gender, introducing the concept of androgyNY. While these studies were executed, major changes took place in the homosexual world, which have been characterized as the masculinization of culture. It is unclear how these changes relate to the masculine and feminine properties of the men involved. Various suggestions are made for further research on the relation between gender and sexual orientation. PMID- 16362242 TI - IQ and posttraumatic stress symptoms in children exposed to interpersonal violence. AB - BACKGROUND: The literature is mixed as to the relationship between intelligence quotient (IQ) and Posttraumatic Stress Disorder (PTSD) symptomatology in adult populations. Even less is known about the relationship in children who have been traumatized. METHODS: Fifty-nine children and adolescents (mean age = 10.6) with a history of interpersonal violence were evaluated with respect to PTSD symptomatology, number of traumas, and estimated Verbal, Performance and Full scale IQ scores. PTSD symptomatology included symptom levels for cluster B (re experiencing), cluster C (avoidance and numbing), and cluster D (Hypervigilance) and criterion F, functional impairment. RESULTS: Results indicated that Full scale and Verbal IQ were significantly associated with the number of traumas, re experiencing symptoms, and impairment. Performance IQ was only associated with impairment. Regression analyses suggested that together PTSD symptomatology predicted Full scale and Verbal IQ but nor Performance IQ and impairment was the single best predictor of IQ generally. CONCLUSIONS: Findings provide support for an association between PTSD symptoms and IQ, particularly verbal IQ. Two possible reasons for this relationship are that higher levels of Verbal IQ may serve as a premorbid protective factor against the development of re-experiencing symptoms, or performance on post-trauma Verbal IQ measures may be negatively impacted by expression of PTSD symptoms. Longitudinal studies are needed to clarify which of these two possibilities explains the association. PMID- 16362241 TI - Perceived neighborhood characteristics and problem behavior among disadvantaged children. AB - Using survey data from former Head Start children in the third grade from 15 sites across the nation (n = 576), this study examines the relationship between maternal subjective neighborhood attributions and their children's behavioral problems. Maternal perceptions of neighborhood characteristics were measured across five domains, including collective efficacy, barriers to services, negative neighbor affects, probability of child status attainment success, and overall neighborhood rating. Children's problem behaviors, measured with the Social Skills Rating System, includes externalizing and internalizing outcomes. Our results suggest that the worse the maternal assessments on each neighborhood construct, the greater the extent of children's problem behavior, holding constant child demographic factors and parental socioeconomic status. In addition, we find that family income effects on children's problem behavior are partially mediated by these perceived neighborhood domains. Taken together, these results suggest that neighborhood deprivation is related to problematic behavioral outcomes in children. PMID- 16362240 TI - A feasibility study on recruiting fathers of young children to examine the impact of paternal depression on child development. AB - Fathers are underrepresented in research on mental health and child outcome. We tested a range of methods of recruitment of fathers to establish feasibility and recruitment rates to obtain a sample for a study on paternal depression and child development. The study took place in North London. Fathers of children aged 6 years and under were approached via: general practice surgeries by post and by face-to-face contact with attendees; child health surveillance clinics face-to face or via their partners and via hospital postnatal wards face-to-face or by post. Researcher time and associated costs were monitored for each method, and symptoms of depression and anxiety measured using the Hospital Anxiety and Depression Scale (HADS). Face-to-face recruitment of fathers on the postnatal ward generated the highest return rate of HADS (28/37; 76%), followed by postal recruitment through general practice (41%;124/303). Lower response rates were observed with postal recruitment via postnatal wards (31%), recruitment in child health surveillance clinics (20%) and approaching attendees in general practice (16%). Twelve percent (23/194) and 30% (58/194) of fathers respectively reported depression and anxiety above the cut-off for case-definition. Costs were calculated on pro-rata researcher time. Costs of recruiting one participant ranged from 3 pounds sterling (general practice postal) to 11.75 pounds sterling (child health surveillance clinics), however the general practice attender method was disproportionately expensive (52.50 pounds sterling). This feasibility study shows that it is possible to recruit fathers to mental health studies and provides clear guidance on planning, costing and the expected levels of recruitment for future studies on fathers of young children. PMID- 16362243 TI - Parental verbal abuse: culture-specific coping behavior of college students in the Philippines. AB - In a larger research study that investigated the conceptualizations, experiences, and coping behaviors of 294 male and female college students exposed to parental verbal abuse, this paper identifies and measures seven culture-specific coping behaviors in the Philippine setting, as experienced by 143 highly abused students identified in the study. Results indicated that both males and females coped essentially through silence. Neither used humor to relieve anxiety nor an intermediary to facilitate communication with the abusive parent. Multivariate analysis of variance suggested that while both experienced anger, females were more inclined than males to accept and tolerate verbal abuse, suppress feelings, and attempt to please the abusive parent. Female victims were disadvantaged due to: differentiated child-rearing expectations and practices; birth order; and the female victim's attitudes, affective reactions, and failure at meaningful communication with the abusive parent. PMID- 16362244 TI - Self-forgiveness, spirituality, and psychological adjustment in women with breast cancer. AB - We evaluated whether a self-forgiving attitude and spirituality were related to psychological adjustment among 81 women being treated for breast cancer at a medical oncology clinic in a county general hospital. Both a self-forgiving attitude and spirituality were unique predictors of less mood disturbance and better quality of life (p's < 0.001). These results are consistent with previous research that has demonstrated a positive relationship between spirituality and well-being. The findings also suggest that self-forgiveness should be explored experimentally to determine whether it can protect against the psychological effects of breast cancer-related stress. Interventions targeting these characteristics could improve the quality of life and alleviate stress, especially in women with breast cancer in public sector settings. PMID- 16362246 TI - Acculturation and sexual function in Asian women. AB - Cultural effects on sexuality are pervasive and potentially of great clinical importance, but have not yet received sustained empirical attention. The purpose of this study was to explore the role of acculturation on sexual permissiveness and sexual function, with a particular focus on arousal in Asian women living in Canada. We also compared questionnaire responses between Asian and Euro-Canadian groups in hopes of investigating whether acculturation captured unique information not predicted by ethnic group affiliation. Euro-Canadian (n = 173) and Asian (n = 176) female university students completed a battery of questionnaires in private. Euro-Canadian women had significantly more sexual knowledge and experiences, more liberal attitudes, and higher rates of desire, arousal, sexual receptivity, and sexual pleasure. Anxiety from anticipated sexual activity was significantly higher in Asian women, but the groups did not differ significantly on relationship satisfaction or problems with sexual function. Acculturation to Western culture, as well as maintained affiliation with traditional Asian heritage, were both significantly and independently related to sexual attitudes above and beyond length of residency in Canada, and beyond ethnic group comparisons. Overall, these data suggest that measurement of acculturation may capture information about an individual's unique acculturation pattern that is not evident when focusing solely on ethnic group comparisons or length of residency, and that such findings may be important in facilitating the assessment, classification, and treatment of sexual difficulties in Asian women. PMID- 16362249 TI - A qualitative study of the relationship between alcohol consumption and risky sex in adolescents. AB - This study examined how alcohol consumption over a single session can affect the likelihood of risky sex in adolescents. Risky sexual behavior, as defined in this study, included sexual intercourse without the use of contraception or aNY sexual behavior that was subsequently regretted. In-depth interviews were conducted with 64 adolescents (aged 14-17 years). Data were analyzed following the principles of Interpretative Phenomenological Analysis. The impact of alcohol consumption upon risky sexual behavior was found to operate through a "continuum of influence." The 5 effects or explanations within this continuum, possibly determined through the level of inebriation, are summarized as follows: (1) alcohol affecting young people's assessment of a person's sexual attractiveness; (2) alcohol used as an "excuse" for socially unacceptable behavior; (3) increased confidence and lowering of inhibitions; (4) impaired judgment in accurately recognizing and controlling a potentially risky situation; and (5) complete loss of control, memory loss, and "black-out." These explanations were ordered with increasing significance. That is, impaired judgment (Explanation 4) and complete loss of control (Explanation 5) arguably represented the most significant effects of drinking alcohol upon risky sexual behavior. For those participants who reported both risky and non-risky sexual behavior, risky events were more likely to occur when alcohol consumption had increased and when the impaired judgment and complete loss of control explanations were applicable. The implications of understanding this complex relationship are outlined, and they will be of interest to academics, practitioners, and policy-makers in the alcohol and sex fields. PMID- 16362248 TI - Validation of the McCoy Female Sexuality Questionnaire in an Italian sample. AB - This article presents the result of a study that translated into Italian and validated the McCoy Female Sexuality Questionnaire (MFSQ) on an Italian sample. The questionnaire was first administered to a sample of 240 Italian women (age range, 18-65 years) recruited from a gynecology clinic. A principal component analysis identified 2 factors: sexuality (9 items) and partnership (5 items). Both factors showed an adequate inter-item reliability (Cronbach's alpha of .88 and .75, respectively). The validity of the Italian MFSQ was then tested by administering the questionnaire to a sample of 16 women with sexual dysfunction and 46 control women. Sexual dysfunction diagnoses were assessed through a semi standardized interview based on the DSM-IV-TR diagnoses for Female Sexual Arousal Disorder, Female Orgasmic Disorder, Dyspareunia, and Hypoactive Sexual Desire Disorder. A discriminant validity test showed significant differences between women with and without female sexual dysfunction. These results indicate that the translated version of the MFSQ is a reliable and valid measure of sexual dysfunction among Italian women. The results also indicated a difference in factor structure between the Italian and the original version of the MFSQ, which warrants further investigation. PMID- 16362251 TI - Implicit attitudes in sexuality: gender differences. AB - This study examined the role of gender in both implicit and explicit attitudes toward sexuality. Implicit attitudes are judgments or evaluations of social objects that are automatically activated, often without the individual's conscious awareness of the causation. In contrast, explicit attitudes are judgments or evaluations that are well established in awareness. As described in Oliver and Hyde's (1993) meta-analysis of self-report (explicit) data, women report greater negative attitudes toward sexuality than do men. In the current study, we used the Sexual Opinion Survey (SOS) developed by Fisher, Byrne, White, and Kelley (1988) to index explicit attitudes and the Implicit Association Test (IAT) developed by Greenwald, McGhee, and Schwartz (1998) to index implicit attitudes. Research has demonstrated that the IAT reveals attitudes that participants may be reluctant to express. Independent variables examined were participant gender, social acceptability of sexual words, and order of associated evaluations in the IAT (switching from positive to negative evaluations or the reverse). The IAT data revealed a significant Order x Gender interaction that showed that women had more negative implicit attitudes toward sexuality than did men. There was also a significant Order x Acceptability interaction, indicating that implicit attitudes were more strongly revealed when the sexual words used in the IAT were more socially unacceptable. As expected, on the SOS, women had more negative explicit attitudes toward sexuality. There was no significant correlation between explicit and implicit attitudes. These data suggest that at both automatic (implicit) and controlled (explicit) levels of attitudes, women harbor more negative feelings toward sex than do men. PMID- 16362250 TI - Sexual regret in college students. AB - A questionnaire study was conducted to assess the relationship between sexual regret and sexual behaviors and demographic factors among 348 college-aged students prior to attending an educational program about sexual health issues. Analyses conducted on the portion of the sample who were sexually active (n = 270) indicated that the majority (71.9%, n = 194) has regretted their decision to engage in sexual activity at least once. The most cited reasons for regret by students included their sexual decision making as inconsistent with their morals (37%), an acknowledgment that alcohol influenced their decision (31.7%), the realization they did not want the same thing as their partner (27.9%), the lack of condom use (25.5%), feeling pressure by their partner (23.0%), and their desire to wait until marriage to have sex (15.4%). The only significant sex difference was that women reported regret due to feeling pressured by a partner more often than men. Multiple and logistic regression analyses indicated that the only significant predictor of regret regarding one's sexual decisions was the number of sexual partners. These results demonstrate the need for sexual educators to incorporate sexual regret into their curricula as the phenomenon of regret is more common than pregnancy and sexually transmitted diseases, the usual focus of sexuality education. PMID- 16362247 TI - Burnout among female indoor sex workers. AB - This study examined levels of "burnout" and associated factors among a sample of female indoor sex workers in the Netherlands (N=96). Levels of burnout on 3 dimensions (emotional exhaustion, depersonalization, and personal competence) were assessed. Only sex workers' mean score on depersonalization was significantly higher than that of a comparison group of female nurses and comparable to those of another comparison group of patients with work-related psychological problems. Evidence was provided for the importance of experiential and context-related factors in burnout among indoor sex workers. For instance, 42% of the variance in depersonalization was explained by not working by choice, negative social reactions, experiences of violence, and lack of control in interaction with clients. Depersonalization may be a strategy to cope with negative conditions and experiences in sex work, but was significantly related to indicators of stress and emotional exhaustion. More than half (53%) of the variance in emotional exhaustion was explained by lack of management support, negative social reactions, not working by choice, and negative working motivation. Personal competence was highest among sex workers with a professional attitude, who started sex work at a relatively older age, and who were well supported by colleagues and management. It is concluded that burnout is not as much associated with sex work per se, but with sex work under certain conditions, among which stigma (notably negative social reactions) and stigma-related experiences (such as role conflict, experiences with violence, and lack of a worker-supportive organizational context) are important. PMID- 16362253 TI - Leuprolide acetate suppresses pedophilic urges and arousability. AB - Cognitive-behavioral psychotherapy was compared with cognitive-behavioral psychotherapy augmented by leuprolide acetate (LA) for suppression of pedophilic behavior. Five male pedophiles (M age, 50 years; range, 36-58) were administered LA by Depo injection for 12 months, followed by saline placebo for 12 months. Testosterone levels, sexual interest preference by visual reaction time (Abel Assessment), penile tumescence (Monarch Penile Plethysmography, PPG), as well as strong sexual urges toward children and masturbatory frequency involving thoughts of children (polygraph), were measured every 3 months. On LA, testosterone decreased to castrate levels. Penile tumescence was significantly suppressed compared with baseline, but sufficient response remained to detect pedophilic interest. Pedophilic interest was also detected by visual reaction times. When asked about having pedophilic urges and masturbating to thoughts of children, all subjects self-reported a decrease. Polygraph responses indicated subjects were not deceptive. On placebo, testosterone and physiologic arousal eventually rose to baseline. As noted by polygraph, at baseline and on placebo, subjects were deceptive regarding increased pedophilic urges and masturbatory frequency. Interest preference, as measured by Abel Assessment and Monarch PPG, was generally unchanged throughout the study. Cognitive-behavioral psychotherapy augmented with LA significantly reduced pedophilic fantasies, urges, and masturbation; however, pedophilic interest did not change during 1 year of therapy. Deceptive responses by polygraph suggested that self-report was unreliable. Follow-up utilizing objective measures is essential for monitoring efficacy of treatment in pedophilia. Our study supports the premise that suppression of pedophilic behavior is possible. LA may augment cognitive behavioral psychotherapy and help break the sequence leading to a re-offense. PMID- 16362252 TI - Sexual and physical health after sex reassignment surgery. AB - A long-term follow-up study of 55 transsexual patients (32 male-to-female and 23 female-to-male) post-sex reassignment surgery (SRS) was carried out to evaluate sexual and general health outcome. Relatively few and minor morbidities were observed in our group of patients, and they were mostly reversible with appropriate treatment. A trend toward more general health problems in male-to females was seen, possibly explained by older age and smoking habits. Although all male-to-females, treated with estrogens continuously, had total testosterone levels within the normal female range because of estrogen effects on sex hormone binding globulin, only 32.1% reached normal free testosterone levels. After SRS, the transsexual person's expectations were met at an emotional and social level, but less so at the physical and sexual level even though a large number of transsexuals (80%) reported improvement of their sexuality. The female-to-males masturbated significantly more frequently than the male-to-females, and a trend to more sexual satisfaction, more sexual excitement, and more easily reaching orgasm was seen in the female-to-male group. The majority of participants reported a change in orgasmic feeling, toward more powerful and shorter for female-to-males and more intense, smoother, and longer in male-to-females. Over two-thirds of male-to-females reported the secretion of a vaginal fluid during sexual excitation, originating from the Cowper's glands, left in place during surgery. In female-to-males with erection prosthesis, sexual expectations were more realized (compared to those without), but pain during intercourse was more often reported. PMID- 16362255 TI - Pure versus co-occurring externalizing and internalizing symptoms in children: the potential role of socio-developmental milestones. AB - Co-occurring internalizing and externalizing disorders are moderately prevalent in children, adolescents, and adults (Anderson, Williams, McGee, & Silva, 1987; McConaughy & Skiba, 1994), but much remains to be understood regarding why some children show "pure" versus co-occurring internalizing and externalizing symptoms. One possible influence that has previously not been considered is the failure to attain socio-developmental milestones, which paradoxically may prevent the development of co-occurring symptoms for some children. The present study proposes a model in which failure to attain relevant socio-developmental milestones might explain why some children may not develop heterotypic co occurring symptoms. Specifically, it is proposed that specific clusters of internalizing symptoms (i.e., high social anxiety, withdrawal, and inhibition) and externalizing symptoms (i.e., high impulsivity, hyperactivity, and emotional reactivity) may be associated with the failure to attain socio-developmental milestones (i.e., poor peer relations for anxious children, lack of self reflection and evaluation for impulsive/reactive children) that, in turn, may prevent subgroups of children from developing co-occurring, heterotypic symptoms. PMID- 16362256 TI - The role of temperament in the etiology of child psychopathology. AB - A substantial proportion of children and adolescents come to suffer from psychological disorders. This article focuses on the temperament factors that are involved in the pathogenesis of child psychopathology. It is argued that besides the reactive temperament factor of emotionality/neuroticism, the regulative process of effortful control also plays an important role in the etiology and maintenance of internalizing and externalizing problems in youths. More specifically, vulnerability to child psychopathology is determined by a temperament that is characterized by high levels of emotionality/neuroticism and low levels of effortful control. Models are hypothesized in which reactive and regulative temperament factors either have interactive or additive effects on the development of psychological disorders in children, and conceptualized in terms of a developmental psychopathology perspective. Directions for future research and clinical implications of this temperamental view on psychopathology are discussed. PMID- 16362257 TI - Child psychopathy: theories, measurement, and relations with the development and persistence of conduct problems. AB - To develop more accurate explanatory and predictive models of child and adolescent conduct problems, interest has grown in examining psychopathic traits in youth. The presence or absence of these traits may help to identify unique etiological pathways in the development of antisocial behavior. The current review provides a detailed summary and analysis of (a) the conceptualization and measurement of child and adolescent psychopathy, (b) research on relations between psychopathic traits and a range of conduct problems and associated variables, (c) efforts to utilize data on child and adolescent psychopathy to delineate a subgroup of youth whose conduct problems share a relatively homogenous etiology and developmental trajectory, and (d) current challenges and limitations in the study of youth psychopathy and directions for future research efforts. Specific sections include the origins of the youth psychopathy construct, conceptualization and measurement, relations between psychopathic traits and conduct problems, relations between measures of psychopathy and measures of individual and contextual variables, efforts to identify a unique subgroup of children with conduct problems using psychopathic traits, and finally, challenges that face the field and goals for the next generation of research. PMID- 16362258 TI - Epidemiological measurement of children's and adolescents' exposure to community violence: working with the current state of the science. AB - A number of measures have been used in epidemiological studies of children's exposure to community violence, yet the quality of these instruments is not uniformly good. This paper undertakes a systematic review of the most commonly used (or most promising) self-report or interview-administered instruments, with regard to their conceptual bases and psychometric properties. Based on the review, recommendations are made for working with the current state of the science in order to move the field forward. A key recommendation is for sounder conceptualization of work in the field and greater transparency in the reporting of research, in order to facilitate the comparability of studies. PMID- 16362259 TI - [Effects of irradiation therapy and inhibiting drugs on the pituitary and its adenomas]. AB - Radiation therapies of pituitary adenomas induce an increase in fibroses and nuclear pleomorphism. Most growth hormone (GH) secreting pituitary adenomas react to somatostatin analogues by a distinct decrease of GH secretion. In two thirds, levels of IGF-1 can be normalized. Some cases show a shrinkage of adenomas that correlates with fibrosis of the tumor. With these drugs, thyroid stimulating hormone secreting adenomas can also be treated. Prolactin secreting adenomas are mostly treated primarily with dopamine agonists. Up to 90% of cases show a strong decrease in hormone secretion and a distinct shrinkage of the adenomas based on strong decrease in adenoma cell volume. Long-term medication with high doses of glucocorticoids induces Crooke's cells in the anterior pituitary. These are suppressed ACTH cells and characterized by increased numbers of large lysosomes and dense bundles of cytofilaments. PMID- 16362260 TI - [Changes in the endometrium after tamoxifen therapy]. AB - Tamoxifen belongs to the group of selective estrogen modulators (SERM) which bind to both the alpha and the beta-estrogen receptors. Depending on the type of tissue, tamoxifen has either an anti-estrogenic or an estrogenic effect on the cells. In the treatment of breast cancer, the anti-estrogenic effect is used. However, at the same time there is a predominant progestin-like and only mild estrogenic effect on the endometrium. Depending on the hormonal situation of the patient, tamoxifen can cause different morphological changes in the endometrium. On ultrasound, these changes are normally diagnosed as a thickening. However, endometrial hyperplasia or endometrial carcinoma is identified histologically in only a few cases. In the majority of cases, the diagnosis is endometrial atrophy or endometrial polyp. Other findings related to tamoxifen therapy are stromal decidualisation, regressive hyperplasia, and foci of mucinous, clear cell and serous metaplasia. The main reason for the diagnosis of endometrial hyperplasia on ultrasound could be fibrosis and edema along the border between the endometrium and myometrium. Still unsolved is the question of whether endometrial carcinomas developing after tamoxifen therapy belong mostly to type I (endometrioid) or type II (serous, clear cell) carcinomas. Only in rare cases do malignant neoplasms other than carcinomas develop after tamoxifen therapy. These are adenosarcomas, carcinosarcomas and endometrial stromal sarcomas. PMID- 16362261 TI - [Cutaneous lesions due to inhibition of epidermal growth factor receptor]. AB - Malignant tumors are increasingly being treated with therapeutic agents having molecular mechanisms of action (so-called biologics). These include therapeutic agents for the blockade of epidermal growth factor receptor (EGFR). The adverse drug reaction profile associated with EGFR inhibitors is dominated by cutaneous lesions. Most common are acneiform skin reactions followed by xerosis, eczema and changes to the hair and nails. The cutaneous changes vary greatly between individuals and may be relatively insignificant. However, they may also prevent continuation of therapy. During the use of EGFR inhibitors, a correlation was observed between the severity of cutaneous changes and the effectiveness of the therapeutic agent, a finding potentially useful for individual dose adjustment. PMID- 16362262 TI - Cyclooxygenase 1 and 2 expressions in the human thyroid gland. AB - We investigated the expression of COX-1 and COX-2 in normal thyroid tissue, follicular adenoma and well-differentiated thyroid carcinomas and evaluated the difference in COX-1 and COX-2 expression. Ten normal thyroid tissues, ten follicular adenomas, ten papillary carcinomas and ten follicular carcinomas were analyzed by semiquantitative reverse transcriptase-polymerase chain reaction (RT PCR) for expression of COX-1 and COX-2 mRNA. In addition, immunohistochemical staining was performed to find the expression of the two enzymes in normal thyroid tissues and thyroid neoplasia. Expression of COX-1 mRNA in the normal thyroid tissues, follicular adenomas and both well-differentiated carcinomas was similar and weak. However, COX-2 mRNA was strongly expressed in the well differentiated carcinomas compared to those of normal thyroid tissue and follicular adenoma. PMID- 16362263 TI - Oncocytic cyst presenting as laryngocele with surgical emphysema. AB - We present a case of laryngeal oncocytic cysts presenting as bilateral laryngoceles and surgical emphysema of the neck. Although there is a rare, but well-documented association of laryngoceles with laryngeal carcinoma, coexistence with another laryngeal pathology is less well reported. PMID- 16362264 TI - Radiofrequency tissue volume reduction of the soft palate and UPPP in the treatment of snoring. AB - The purpose of this study was to evaluate the efficiency of radiofrequency tissue volume reduction (RFTVR) and uvulopalatopharyngoplasty (UPPP) in the treatment of snoring in a prospective clinical trial of 79 patients consecutively undergoing surgery for snoring. Seventy-nine patients with primary snoring or mild OSAS (obstructive sleep apnea syndrome) were enrolled in this clinical trial (66 males and 13 females). According to the anatomical findings (the size of the tonsils and uvula), the patients underwent UPPP/TE of the RFTVR of the soft palate. Forty seven patients had UPPP/TE (age 45.81+/-12.11 years; median AHI: 8; range 1-29). Thirty-two patients were treated with RFTVR of the soft palate (age 48.10+/ 10.92; median AHI: 5.0; range 0-26). The average number of treatments was 2.2. All patients underwent preoperative polysomnography to exclude severe OSAS. Pre- and postoperative snoring scores were evaluated from the patients with bed partners. Postoperative follow-up data were collected at a median of 4 months after treatment; 85.1% of the UPPP group and 53.1% of the RFTVR group underwent postoperative polysomnography. Subjective snoring scores of all study participants were evaluated. Preoperatively, there was no statistically significant difference of subjective symptoms, age and BMI between the two groups. The snoring scores improved statistically significantly in both groups (P <0.001 in the UPPP group; P =0.001 in the RFTVR group). After UPPP/TE snoring improved in 37 patients (78.7%), and 29 (61.7%) thereof were free of bothersome snoring; no change was found in 9 patients (19.2%), and 1 (2.1%) worsened. In the RFTVR group, snoring improved in 15 (46.9%), and 9 (28.1%) thereof were free of bothersome snoring; no change was found in 13 patients (50%), and 1 worsened (3.1%). Preoperative AHI was statistically higher (P =0.016) and mean minimal oxygen saturation significantly lower (P =0.002) in the UPPP group. In the UPPP group AHI and HI showed statistically significant improvement postoperatively (P =0.025 and P =0.034, respectively). After RFTVR, no statistically significant change of AHI, HI or oxygen saturation was found. Besides limited mucosal erosions (15%) after RFTVR and foreign body sensations (<10%) after UPPP/TE, no side effects were observed. The success rate of RFTVR of the soft palate is lower compared to the more invasive technique of UPPP. Due to its minimally invasive character, RFTVR is suitable as first-step treatment for snoring, but patients should be counseled about possible success rates and different treatment options. PMID- 16362265 TI - Microvascular transplantation and replantation of the dog submandibular gland. AB - Transplantation and replantation of the submandibular gland with microvascular techniques were demonstrated in a previous study, with good gland survival. The application of radiation on the neck bed was attempted to address an actual clinical scenario in this study. Five canine submandibular glands were transplanted using microvascular techniques to the ipsilateral femoral system. Radiotherapy at a dosage level of 3,600 cGy using 600 cGy q.d was delivered to the nasopharyngeal and neck regions 2 weeks after transplantation. The transferred glands were then reintroduced into the original but radiated neck bed. The glands were harvested for histological examination 8 weeks later. Four of five canine submandibular glands can withstand microvascular transplantation and then replantation into a radiated neck bed for at least 8 weeks. However, the salivary function was depleted. The canine submandibular gland can survive the transplantation and replantation for at least 8 weeks in spite of precipitating radiation insult on the neck bed for 3 weeks. Neurorraphy is, however, essential to maintaining the glandular function. PMID- 16362267 TI - Inflammatory focal myositis of the sternomastoid muscle: is there an absolute indication for biopsy? A case report and review of the literature. AB - Focal myositis is a localised inflammatory process affecting skeletal muscles belonging to the pathological group of inflammatory pseudo tumours of soft tissue that includes myositis ossificans, proliferative myositis and nodular pseudosarcomatous fasciitis. Very rarely, it may affect one of the neck muscles and present as a neck lump, in which case both the clinical and pathological picture can mimic a sarcoma. We describe a case of focal myositis of the sternocleidomastoid muscle, present a review of this rare condition and debate the necessity of biopsy. PMID- 16362266 TI - Defensin and chemokine expression patterns in the palatine tonsil: a model of their local interaction. AB - Defensins and chemokines are an essential part of the immune response mechanisms in the head and neck mucosa. This work investigates their correlation and their expression pattern in tonsillar disease. Forty-four tonsil tissue samples were obtained from patients who underwent tonsillectomy between 1998 and 1999 for chronic tonsillitis with (n =9) and without (n =25) inflammatory infiltrates and hyperplasia of the tonsil (n =10). Defensin (hBD-1, hBD-2, HNP-1 and HNP-4) and chemokine (RANTES, eotaxin, eotaxin-2, MCP-3, MCP-4 and IL-8) mRNA expressions were analyzed by SQRT-PCR. HNP-4 and eotaxin-2 expressions were positively correlated (P <0.05) in the acute tonsillitis group. HBD-2 and MCP-3 expressions were positively correlated in the hyperplastic tonsils group. Within all groups together, HNP-4 and RANTES expressions were highly positively correlated (P <0.01), and HNP-1 and hBD-2 were positively correlated with IL-8 expressions. Immunohistochemistry demonstrated eotaxin-1 as well as IL-8 production to be predominantly located within the lymphoid follicles and submucosa. RANTES production was shown in the epithelial lining and perivascular tissue. The expression of hBD-1 and hBD-2 was limited to the epithelial lining. Our data support an association between the innate and acquired immune systems on the defensin-chemokine level. The finding of positively correlated hBD-2 and IL-8 expression is biologically relevant because of the proximity of hBD-2 (epithelium) and IL-8 (submucosa) release, as well as the synergistic support of the Th1 system. In addition, our data suggest RANTES as a first-line mediator of perivascular leukocyte recruitment. PMID- 16362268 TI - Extended and standard supraglottic laryngectomies: a review of 110 patients. AB - The purpose of this study was to compare functional and oncological results of extended and standard supraglottic laryngectomies. One hundred ten patients with supraglottic carcinoma were treated. A standard supraglottic laryngectomy (SSL), a laterally extended supraglottic laryngectomy (LESL) and an anteriorly extended supraglottic laryngectomy (AESL) were performed on 32, 47 and 31 patients, respectively. Indications for postoperative radiotherapy included positive surgical margins (23% of patients) and/or node metastasis (63% of patients). Local recurrence occurred in 13% of SSL, 15% of LESL and 17% of AESL patients. Pulmonary complications due to aspiration were observed in 6% of SSL, 15% of LESL and 19% of AESL. The overall 5-year cure rates were 63% for SSL, 45% for LESL and 47% for the AESL procedures. Extended supraglottic laryngectomies provided as good a local tumor control as SSL. Extension to the hypopharynx (LESL) and to the vallecula (AESL) showed more frequent pulmonary complications and reduced cure rates. PMID- 16362270 TI - Lowering the thyroid dose in screening examinations of the cervical spine. AB - The first objective of this study was to test the hypothesis that a lower-dose (14.1 mGy thyroid dose) protocol for helical computed tomography (CT) of the entire cervical spine demonstrates equivalent technical adequacy and diagnostic accuracy as the standard-dose protocol (26.0 mGy thyroid dose) used at our institution. The second objective was to estimate the excess thyroid cancer mortality for three cervical spine screening protocols. Eight patients underwent two helical CT acquisitions of the entire cervical spine (standard and lower dose); from these acquisitions, a database of 128 randomized images (64 standard dose and 64 lower dose) was constructed. Three radiologists evaluated each of the 128 images for technical adequacy and, if the image was technically adequate, diagnostic accuracy. Historical data of excess thyroid cancer mortality stratified by age and sex were used to estimate the impact of lowering the thyroid dose in cervical spine screening. Estimates used a linear extrapolation of mortality data. The lower-dose protocol for helical CT of the entire cervical spine demonstrates equivalent technical adequacy and diagnostic accuracy as the standard protocol. The excess thyroid cancer mortality is a function of patient age and sex; for 25-year-old men, the excess mortality per 100,000 patients is 96.7 (standard-dose CT), 52.4 (lower-dose CT), and 6.7 (radiographs alone, 1.8 mGy thyroid dose). The equivalent technical adequacy and diagnostic accuracy of a lower-dose protocol for helical CT of the entire cervical spine support its implementation in routine screening. The excess thyroid mortality emphasizes the need to maintain an open dialogue with our referring clinicians with respect to the mechanism of injury, clinical findings, and radiation risks. PMID- 16362271 TI - Consecutive instances of gallstone ileus due to obstruction first at the ileum and then at the duodenum complicating a gallbladder carcinoma: a case report. AB - Ectopic gallstone obstruction, gallstone ileus, due to cholecystoenteric fistula is an infrequent condition. Its occurrence as a complication of a gallbladder (GB) carcinoma is even more rare. We describe an unusual case of a GB carcinoma complicated by a cholecystoduodenal fistula leading to first gallstone obstruction in the ileum and then, later, in the duodenum, in which an accurate preoperative diagnosis was based on pathognomonic computerized tomography (CT) features. A correct diagnosis of this may be crucial and requires prompt surgical intervention; radiologists should be familiar with their classic CT appearance. PMID- 16362272 TI - Destruction of Deinococcus geothermalis biofilm by photocatalytic ALD and sol-gel TiO2 surfaces. AB - The aim of the present work was to explore possibilities of photocatalytic TiO2 coating for reducing biofilms on non-living surfaces. The model organism, Deinococcus geothermalis, known to initiate growth of durable, colored biofilms on machine surfaces in the paper industry, was allowed to form biofilms on stainless steel, glass and TiO2 film coated glass or titanium. Field emission electron microscopy revealed that the cells in the biofilm formed at 45 degrees C under vigorous shaking were connected to the surface by means of numerous adhesion threads of 0.1-0.3 microm in length. Adjacent cells were connected to one another by threads of 0.5-1 microm in length. An ultrastructural analysis gave no indication for the involvement of amorphous extracellular materials (e.g., slime) in the biofilm. When biofilms on photocatalytic TiO2 surfaces, submerged in water, were exposed to 20 W h m(-2) of 360 nm light, both kinds of adhesion threads were completely destroyed and the D. geothermalis cells were extensively removed (from >10(7) down to below 10(6) cells cm(-2)). TiO2 films prepared by the sol-gel technique were slightly more effective than those prepared by the ALD technique. Doping of the TiO2 with sulfur did not enhance its biofilm-destroying capacity. The results show that photocatalytic TiO2 surfaces have potential as a self-cleaning technology for warm water using industries. PMID- 16362273 TI - Molecular analysis of the high stearic acid content in sunflower mutant CAS-14. AB - Increasing the stearic acid content to improve sunflower (Helianthus annuus L.) oil quality is a desirable breeding objective for food-processing applications. CAS-14 is a sunflower mutant line with a high stearic acid content in its seed oil (>35% vs. <6% in currently grown sunflower hybrids), which is controlled by the Es3 gene. However, the expression of the high stearic acid character in CAS 14 is strongly influenced by temperature during seed maturation and it is not uniform along the seed. The objectives of this study were (1) to identify PCR based molecular markers linked to the Es3 gene from CAS-14, (2) to map this gene on the sunflower genetic map, and (3) to characterize the interaction between CAS 14 and CAS-3, a sunflower high stearic acid (about 26%) mutant line with the Es1 and Es2 genes determining this trait. Two F2 mapping populations were developed from crosses between CAS-14 and P21, a nuclear male sterile line with the Ms11 gene controlling this character, and between CAS-14 and CAS-3. One hundred and thirty-three individuals from P21xCAS-14, and 164 individuals from CAS-3xCAS-14 were phenotyped in F2 and F3 seed generations for fatty acid composition using gas-liquid chromatography, and they were then genotyped with microsatellite [simple sequence repeat (SSR)] and insertion-deletion (INDEL) markers. Bulk segregant analysis in the P21xCAS-14 population identified two markers on LG 8 putatively linked to Es3. A large linkage group was identified using additional markers mapping to LG 8. Es3 mapped to the distal half of LG 8 and was flanked by the SSR markers ORS243 and ORS1161 at genetic distances of 0.5, and 3.9 cM, respectively. The Ms11 gene was also mapped to LG 8 and genetic distance between this gene and Es3 was found to be 7.4 cM. In the CAS-3xCAS-14 population, two QTLs were identified on LG 1 and LG 8, which underlie the Es1 gene from CAS-3 and the Es3 gene from CAS-14, respectively. A significant epistatic interaction between these two QTLs was found. Results from this study provided a basis for determining CAS-14 efficient breeding strategies. PMID- 16362274 TI - Identification and mapping of microsatellite markers linked to a root-knot nematode resistance gene (rkn1) in Acala NemX cotton (Gossypium hirsutum L.). AB - Host-plant resistance is the most economic and effective strategy for root-knot nematode (RKN) Meloidogyne incognita control in cotton (Gossypium hirsutum L.). Molecular markers linked to resistance are important for incorporating resistance genes into elite cultivars. To screen for microsatellite markers (SSR) closely linked to RKN resistance in G. hirsutum cv. Acala NemX, F1, F2, BC1F1, and F2:7 recombinant inbred lines (RILs) from intraspecific crosses and an F2 from an interspecific cross with G. barbadense cv. Pima S-7 were used. Screening of 284 SSR markers, which cover all the known identified chromosomes and most linkage groups of cotton, was performed by bulked segregant analysis, revealing informative SSRs. The informative SSRs were then mapped on the above populations. One co-dominant SSR marker CIR316 was identified tightly linked to a major resistance gene (designated as rkn1), producing amplified DNA fragments of approximately 221 bp (CIR316a) and 210 bp (CIR316c) in Acala NemX and susceptible Acala SJ-2, respectively. The linkage between CIR316a marker and resistance gene rkn1 in Acala NemX had an estimated distance of 2.1-3.3 cM depending on the population used. Additional markers, including BNL1231 with loose linkage to rkn1 (map distance 25.1-27.4 cM), BNL1066, and CIR003 allowed the rkn1 gene to be mapped to cotton linkage group A03. This is the first report in cotton with a closely linked major gene locus determining nematode resistance, and informative SSRs may be used for marker-assisted selection. PMID- 16362275 TI - Identification of Glu-B1-1 as a candidate gene for the quantity of high-molecular weight glutenin in bread wheat (Triticum aestivum L.) by means of an association study. AB - A previous study in wheat (Triticum aestivum L.) identified two candidate genes controlling a quantitative trait locus (QTL) for high-molecular-weight glutenin subunit (HMW-GS) GluBx. These candidates were Glu-B1-1, the structural gene coding for Glu1Bx, and the B homoeologous gene coding for SPA (spa-B), a seed storage protein activator. The goal of this study was to identify the best candidate gene for this QTL. Single nucleotide polymorphisms (SNPs) are an abundant source of DNA polymorphisms that have been successfully used to identify loci associated with particular phenotypes. As no linkage disequilibrium was detected between Glu-B1-1 and spa-B, we performed an association study to identify the individual gene responsible for the QTL. Six SNPs, three located in Glu-B1-1 and three in spa-B, were genotyped by mass spectrometry in a collection of 113 bread wheat lines. These lines were also evaluated for protein content as well as the total quantity of HMW-GSs and of each HMW-GS in seed samples from two harvest years. Significant associations were detected only between Glu-B1-1 polymorphism and most of the traits evaluated. Spa-B was unambiguously discarded as a candidate. To our knowledge, this is the first report on an association study that was successfully used to discriminate between two candidate genes. PMID- 16362276 TI - Quantitative trait loci for cell wall components in recombinant inbred lines of maize (Zea mays L.) II: leaf sheath tissue. AB - While maize silage is a significant feed component in animal production operations, little information is available on the genetic bases of fiber and lignin concentrations in maize, which are negatively correlated with digestibility. Fiber is composed largely of cellulose, hemicellulose and lignin, which are the primary components of plant cell walls. Variability for these traits in maize germplasm has been reported, but the sources of the variation and the relationships between these traits in different tissues are not well understood. In this study, 191 recombinant inbred lines of B73 (low-intermediate levels of cell wall components, CWCs) x De811 (high levels of CWCs) were analyzed for quantitative trait loci (QTL) associated with CWCs in the leaf sheath. Samples were harvested from plots at two locations in 1998 and one in 1999 and assayed for neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL). QTL were detected on all ten chromosomes, most in tissue specific clusters in concordance with the high genotypic correlations for CWCs within the same tissue. Adjustment of NDF for its subfraction, ADF, revealed that most of the genetic variation in NDF was probably due to variation in ADF. The low to moderate genotypic correlations for the same CWC across leaf sheath and stalk tissues indicate that some genes for CWCs may only be expressed in certain tissues. Many of the QTL herein were detected in other populations, and some are linked to candidate genes for cell wall carbohydrate biosynthesis. PMID- 16362277 TI - Stacking quantitative trait loci (QTL) for Fusarium head blight resistance from non-adapted sources in an European elite spring wheat background and assessing their effects on deoxynivalenol (DON) content and disease severity. AB - Fusarium head blight (FHB) is a devastating disease in wheat that reduces grain yield, grain quality and contaminates the harvest with deoxynivalenol (DON). As potent resistance sources Sumai 3 and its descendants from China and Frontana from Brazil had been analysed by quantitative trait loci (QTL) mapping. We introgressed and stacked two donor QTL from CM82036 (Sumai 3/Thornbird) located on chromosomes 3B and 5A and one donor QTL from Frontana on chromosome 3A in elite European spring wheat and estimated the effects of the three individual donor QTL and their four combinations on DON, Fusarium exoantigen content, and FHB rating adjusted to heading date. One class with the susceptible QTL alleles served as control. Each of the eight QTL classes was represented by 12-15 F(3) derived lines tested in F(5) generation as bulked progeny possessing the respective marker alleles homozygously. Traits were evaluated in a field experiment across four locations with spray inoculation of Fusarium culmorum. All three individual donor-QTL alleles significantly reduced DON content and FHB severity compared to the marker class with no donor QTL. The only exception was the donor-QTL allele 3A that had a low, but non-significant effect on FHB severity. The highest effect had the stacked donor-QTL alleles 3B and 5A for both traits. They jointly reduced DON content by 78% and FHB rating by 55% compared to the susceptible QTL class. Analysis of Fusarium exoantigen content illustrates that lower disease severity is associated with less mycelium content in the grain. In conclusion, QTL from non-adapted sources could be verified in a genetic background of German elite spring wheat. Within the QTL classes significant (P<0.05) genotypic differences were found among the individual genotypes. An additional phenotypic selection would, therefore, be advantageous after performing a marker-based selection. PMID- 16362278 TI - Manifestation of heterosis during early maize (Zea mays L.) root development. AB - Heterosis is typically detected in adult hybrid plants as increased yield or vigor compared to their parental inbred lines. Only little is known about the manifestation of heterosis during early postembryonic development. Objective of this study was to identify heterotic traits during early maize root development. Four German inbred lines of the flint (UH002 and UH005) and dent (UH250 and UH301) pool and the 12 reciprocal hybrids generated from these inbred lines were subjected to a morphological and histological analysis during early root development. Primary root length and width were measured daily in a time course between 3 and 7 days after germination (DAG) and displayed average midparent heterosis (MPH) of 17-25% and 1-7%, respectively. Longitudinal size of cortical cells in primary roots was determined 5 DAG and displayed on average 24% MPH thus demonstrating that enlarged primary roots of hybrids can mainly be attributed to elongated cortical cells. The number of seminal roots determined 14 DAG showed on average 18% MPH. Lateral root density of all tested hybrids was determined 5 DAG. This root trait showed the highest degree of heterosis with an average MPH value of 51%. This study demonstrated that heterosis is already manifesting during the very early stages of root development a few days after germination. The young root system is therefore a suitable model for subsequent molecular studies of the early stages of heterosis manifestation during seedling development. PMID- 16362279 TI - Can we slow the rising incidence of childhood-onset autoimmune diabetes? The overload hypothesis. AB - Overload of the beta cell, mediated by a variety of mechanisms, may sensitise it to immune damage and apoptosis, and thus accelerate ongoing autoimmune processes leading to its destruction. Environmental risk determinants that may exert such overload effects include insulin resistance due to excess fat cell accumulation, and increased insulin requirement due to a high growth rate, physical stress (infection, inflammation) or psychological stress. The increasing incidence of childhood diabetes, and the shift to younger age at onset, is unlikely to be driven by environmental risk factors that have been associated with initiation of autoimmunity, e.g. virus infections or early infant feeding. Risk factors that may accelerate beta cell destruction have shown a steady increase in the population, and are more plausible causes of such a pattern of change. Child growth, weight and birthweight are well-established estimates of community wealth and increase in most countries of Europe. Overfeeding of children early in life leads to both accelerated growth and weight, and even a moderate excess of child growth, not necessarily associated with obesity, is associated with risk of type 1 diabetes. New, safe and effective immune-modulating drugs for possible arrest of the autoimmune process may become available in time, but in the interim these accelerating factors may be targeted. Public health programmes for pregnant mothers and young families, aiming at changing overfeeding and the sedentary lifestyle of the children would be preferable to other alternatives. Interventions such as these would be safe and could potentially influence future risks of type 1 and type 2 diabetes and other major threats to adult health. PMID- 16362280 TI - Acute in vivo effects of insulin on gene expression in adipose tissue in insulin resistant and insulin-sensitive subjects. AB - AIMS/HYPOTHESIS: We determined the response of selected genes to in vivo insulin in adipose tissue in 21 non-diabetic women. MATERIALS AND METHODS: The women were divided into insulin-sensitive and -resistant groups based on their median whole body insulin sensitivity (8.7+/-0.4 vs 4.2+/-0.3 mg kg(-1) min(-1) for insulin sensitive vs -resistant group). Subcutaneous adipose tissue biopsies were obtained before and after 3 and 6 h of i.v. maintained euglycaemic hyperinsulinaemia. Adipose tissue mRNA concentrations of facilitated glucose transporter, member 1 (SLC2A1, previously known as GLUT1), facilitated glucose transporter, member 4 (SLC2A4, previously known as GLUT4), peroxisome proliferator-activated receptor gamma ( PPARG), peroxisome proliferator-activated receptor gamma co-activator 1alpha (PPARGC1A), 11beta-hydroxysteroid dehydrogenase-1 (HSD11B1), TNF, adiponectin (ADIPOQ), IL6 and the macrophage marker CD68 were measured using real-time PCR. RESULTS: Basal expression of 'insulin-sensitivity genes' SLC2A4 and ADIPOQ was lower while that of 'insulin resistance genes', HSD11B1 and IL6 was significantly higher in the insulin resistant than in the insulin-sensitive group. Insulin significantly increased expression of 'insulin-sensitivity genes' SLC2A4, PPARG, PPARGC1A and ADIPOQ in the insulin-sensitive group, while only expression of PPARG and PPARGC1A was increased in the insulin-resistant group. The expression of 'insulin-resistance genes' HSD11B1 and IL6 was increased by insulin in the insulin-resistant group, but insulin failed to increase HSD11B1 expression in the insulin-sensitive group. At 6 h, expression of HSD11B1, TNF and IL6 was significantly higher in the insulin-resistant than in the insulin-sensitive group. IL6 expression increased significantly more in response to insulin in the insulin-resistant than in the insulin-sensitive group. CD68 was overexpressed in the insulin-resistant as compared with the insulin-sensitive group at both 0 and 6 h. CONCLUSIONS/INTERPRETATION: These data suggest that genes adversely affecting insulin sensitivity hyperrespond to insulin, while genes enhancing insulin sensitivity hyporespond to insulin in insulin-resistant human adipose tissue in vivo. PMID- 16362281 TI - Troglitazone: the lesson that nobody learned? PMID- 16362282 TI - Comment on: Biason-Lauber A, Boehm B, Lang-Muritano M, et al (2005) Association of childhood type 1 diabetes mellitus with a variant of PAX4: possible link to beta cell regenerative capacity. Diabetologia 48:900-905. PMID- 16362283 TI - Glucagon secretion in relation to insulin sensitivity in healthy subjects. AB - AIMS/HYPOTHESIS: The study evaluated whether glucagon secretion is regulated by changes in insulin sensitivity under normal conditions. MATERIALS AND METHODS: A total of 155 healthy women with NGT (aged 53-70 years) underwent a glucose dependent arginine-stimulation test for evaluation of glucagon secretion. Arginine (5 g) was injected i.v. under fasting conditions (plasma glucose 4.8+/ 0.1 mmol/l) and after raising blood glucose concentrations to 14.8+/-0.1 and 29.8+/-0.2 mmol/l. The acute glucagon response (AGR) to arginine during the three glucose levels (AGR(1), AGR(2), AGR(3)) was estimated, as was the suppression of baseline glucagon by the increased glucose. All women also underwent a 2-h euglycaemic-hyperinsulinaemic clamp study for estimation of insulin sensitivity. RESULTS: Insulin sensitivity was normally distributed, with a mean of 73.2+/-29.3 (SD) nmol glucose kg(-1) min(-1)/pmol insulin l(-1). When relating the variables obtained from the arginine test to insulin sensitivity, insulin resistance was associated with increased AGR and with increased suppression of glucagon levels by glucose. For example, the regression between insulin sensitivity and AGR(2) was r=-0.38 (p<0.001) and between insulin sensitivity and suppression of glucagon levels by 14.8 mmol/l glucose r=0.36 (p<0.001). Insulin sensitivity also correlated negatively with insulin secretion; multivariate analysis revealed that changes in insulin sensitivity and insulin secretion were independently related to changes in glucagon secretion. CONCLUSIONS/INTERPRETATION: The body adapts to insulin resistance by increasing the glucagon response to arginine and by increasing the suppression of glucagon levels by glucose. Hence, not only the islet beta cells but also the alpha cells seem to undergo compensatory changes during the development of insulin resistance. PMID- 16362284 TI - Insulin resistance causes increased beta-cell mass but defective glucose stimulated insulin secretion in a murine model of type 2 diabetes. AB - AIMS/HYPOTHESIS: Although insulin resistance induces compensatory increases in beta cell mass and function to maintain normoglycaemia, it is not clear whether insulin resistance can precipitate beta cell dysfunction and hyperglycaemia without a pre-existing beta cell susceptibility. We therefore examined the beta cell phenotype in the MKR mouse, a model in which expression of a dominant negative IGF 1 receptor (IGF1R) in skeletal muscle leads to systemic insulin resistance and diabetes. MATERIALS AND METHODS: Circulating glucose, insulin and glucagon concentrations were measured. Insulin sensitivity, glucose tolerance and insulin release in vivo were assessed by i.p. insulin and glucose tolerance tests. Beta cell function was assessed via insulin secretion from isolated islets and the glucose gradient in the perfused pancreas. Beta cell morphology was examined via immunohistochemistry. MKR mice were fed a high-fat diet containing sucrose (HFSD) to test metabolic capacity and beta cell function. RESULTS: Insulin-resistant MKR mice developed hyperglycaemia and a loss of insulin responsiveness in vivo. Basal insulin secretion from the perfused pancreas was elevated, with no response to glucose. Despite the demand on insulin secretion, MKR mice had increased pancreatic insulin content and beta cell mass mediated through hyperplasia and hypertrophy. The HFSD worsened hyperglycaemia in MKR mice but, despite increased food intake in these mice, failed to induce the obesity observed in wild-type mice. CONCLUSIONS/INTERPRETATION: Our studies demonstrate that insulin resistance of sufficient severity can impair glucose-stimulated insulin secretion, thereby undermining beta cell compensation and leading to hyperglycaemia. Moreover, because insulin stores were intact, the secretory defects reflect an early stage of beta cell dysfunction. PMID- 16362285 TI - The in vivo effects of the Pro12Ala PPARgamma2 polymorphism on adipose tissue NEFA metabolism: the first use of the Oxford Biobank. AB - AIMS/HYPOTHESIS: To investigate the phenotypic effects of common polymorphisms on adipose tissue metabolism and cardiovascular risk factors, we set out to establish a biobank with the unique feature of allowing a prospective recruit-by genotype approach. The first use of this biobank investigates the effects of the peroxisome proliferator-activated receptor (PPAR) Pro12Ala polymorphism on integrative tissue-specific physiology. We hypothesised that Ala12 allele carriers demonstrate greater adipose tissue metabolic flexibility and insulin sensitivity. MATERIALS AND METHODS: From a comprehensive population register, subjects were recruited into a biobank, which was genotyped for the Pro12Ala polymorphism. Twelve healthy male Ala12 carriers and 12 matched Pro12 homozygotes underwent detailed physiological phenotyping using stable isotope techniques, and measurements of blood flow and arteriovenous differences in adipose tissue and muscle in response to a mixed meal containing [1,1,1-(13)C]tripalmitin. RESULTS: Of 6,148 invited subjects, 1,072 were suitable for inclusion in the biobank. Among Pro12 homozygotes, insulin sensitivity correlated with HDL-cholesterol concentrations, and inversely correlated with blood pressure, apolipoprotein B, triglyceride and total cholesterol concentrations. Ala12 carriers showed no such correlations. In the meal study, Ala12 carriers had lower plasma NEFA concentrations, higher adipose tissue and muscle blood flow, and greater insulin mediated postprandial hormone-sensitive lipase suppression along with greater insulin sensitivity than Pro12 homozygotes. CONCLUSIONS/INTERPRETATION: This study shows that a recruit-by-genotype approach is feasible and describes the biobank's first application, providing tissue-specific physiological findings consistent with the epidemiological observation that the PPAR Ala12 allele protects against the development of type 2 diabetes. PMID- 16362286 TI - Ultrastructural organization of Alicyclobacillus tolerans strain K1T cells. AB - Electron microscopy examinations of thin sections and freeze-fracture replicas revealed the specific ultrastructural features of Alicyclobacillus tolerans strain K1(T). In particular, the cell wall displayed an ultrastructure typical of gram-positive bacteria and consisted of a thin murein layer (50-60 A in thickness); cells exhibited a surface S-layer constituted by large hexagonally packed (p6-symmetry) rod-shaped subunits of 150-160 A in diameter and 200 A in height. In the cytoplasmic membrane, there were intramembrane vesicular structures that sometimes appeared as large leaflets in the central part. The cytoplasm contained numerous vesicular inclusions covered with a monolayered wall, dissimilar to bilamellar lipid membranes. Endospore coats displayed an intricate structure and consisted of three thick layers; the outer layer had an unusual fine structure; the exosporium was also found. PMID- 16362287 TI - Structural modeling and environmental regulation of arginine decarboxylase in Synechocystis sp. PCC 6803. AB - Arginine decarboxylase (ADC) is the first enzyme in the alternative route to putrescine in the polyamine biosynthesis pathway in bacteria and plants. In this study, we have focused on the effects of various types of short-term stresses on the transcript amount and specific activity of Synechocystis sp. PCC 6803 ADC. Our results reveal that the steady-state transcript accumulation and enzyme activity are not connected in a simple manner, since only photoheterotrophy and synergistic salt and high-light stress affected both parameters similarly. Changes in the steady-state ADC mRNA accumulation under the other short-term stress conditions studied had only a small impact on enzyme activity, suggesting post-translational regulation. Based on structural modeling, Synechocystis ADCs have a putative extra domain, which might be involved in the post-translational regulation of ADC activity in Synechocystis. In addition, two symmetric inter subunit disulfide bonds seem to stabilize the dimeric structure of ADCs. There are two genes coding for ADC and agmatinase, another polyamine pathway enzyme, in Synechocystis genome, while the genes coding for ornithine decarboxylase and for some other enzymes in the polyamine pathway were not identified with homology searches. PMID- 16362288 TI - Physiological role of D-amino acid-N-acetyltransferase of Saccharomyces cerevisiae: detoxification of D-amino acids. AB - Saccharomyces cerevisiae is sensitive to D-amino acids: those corresponding to almost all proteinous L-amino acids inhibit the growth of yeast even at low concentrations (e.g. 0.1 mM). We have determined that D-amino acid-N acetyltransferase (DNT) of the yeast is involved in the detoxification of D-amino acids on the basis of the following findings. When the DNT gene was disrupted, the resulting mutant was far less tolerant to D-amino acids than the wild type. However, when the gene was overexpressed with a vector plasmid p426Gal1 in the wild type or the mutant S. cerevisiae as a host, the recombinant yeast, which was found to show more than 100 times higher DNT activity than the wild type, was much more tolerant to D-amino acids than the wild type. We further confirmed that, upon cultivation with D-phenylalanine, N-acetyl-D-phenylalanine was accumulated in the culture but not in the wild type and hpa3Delta cells overproducing DNT cells. Thus, D-amino acids are toxic to S. cerevisiae but are detoxified with DNT by N-acetylation preceding removal from yeast cells. PMID- 16362289 TI - Fabrication and characterization of stable ultrathin film micropatterns containing DNA and photosensitive polymer diazoresin. AB - Stable, ultrathin DNA micropatterns were fabricated from photosensitive polymer diazoresin (DR) through a self-assembly technique. The micropatterns were achieved on LBL ultrathin film after UV exposure through a photomask. The patterns were characterized systematically with scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy and fluorescence microscopy. All of the results indicate that the combined LBL self-assembly and photolithography technique is a promising method for constructing stable, well defined micropatterns with a nanoscale thickness. PMID- 16362290 TI - Simultaneous optimization of resolution and analysis time in mixed micellar liquid chromatography of coumarins by use of a utility function. AB - A chemometrics approach has been used to optimize the separation of eight coumarin compounds by mixed micellar liquid chromatography. A utility function, a multi-criterion decision-making (MCDM) method, was tested for evaluation of two different measures of chromatographic performance (resolution and analysis time). The effect of six experimental parameters on a chromatographic response function (CRF) was investigated. The factors studied were the concentrations of SDS and Brij-35, alkyl-chain length of the alcohol used as organic modifier, organic modifier concentration, mobile phase pH, and temperature. The experiments were performed according to a face-centered cube response-surface experimental design. For evaluation of the chromatograms a simple linear response function was used which expressed as a summation of two optimization criteria, resolution and analysis time. Then calculated CRF values were fitted to a polynomial model to correlate the CRF values with the variables and their interactions. The regression model obtained was characterized by both its descriptive and predictive ability (R=0.963 and R2cv=0.861) and used, by means of a grid-search algorithm, to optimize the chromatographic conditions. Experiments performed under the optimum conditions predicted by the model produced a chromatogram of high quality. The model was also verified by the good agreement observed between predicted and experimental values of the chromatographic response function under the optimum conditions. PMID- 16362291 TI - Comparison of different mass spectrometric detection techniques in the gas chromatographic analysis of pyrethroid insecticide residues in soil after microwave-assisted extraction. AB - A comparative study was carried out for the simultaneous determination of 11 pyrethroid insecticides in soil by gas chromatography (GC)--ion trap (IT)--mass spectrometry (MS), by means of two different ionization modes; electron impact and negative chemical ionization and three data acquisition procedures; full scan, selected ion monitoring and MS/MS. Pyrethroids investigated were tetramethrin, bifenthrin, phenothrin, lambda-cyhalothrin, permethrin, cyfluthrin, cypermethrin, flucythrinate, esfenvalerate, fluvalinate and deltamethrin. Soil samples were treated with toluene/water by microwave-assisted extraction for 9 min at 700 W and a cleaning up with florisil was performed. Clean soil samples were spiked with pyrethroids at a spiking level of 10, 25 and 50 ng/g. The method employed provides a concentration factor of 10. The ionization gas employed in the negative chemical ionization mode was methane. The use of MS/MS acquisition, in electron impact ionization, provided the best results, due to its high selectivity and sensitivity, giving very low limits of detection from 0.08 to 0.54 ng/g. In negative chemical ionization full scan and selected ion monitoring methods detection limits from 0.12 to 1.40 ng/g were found. The proposed methods were applied to several levels from 10 to 50 ng/g of spiked soils, being electron impact MS/MS method which minimizes matrix spectrum interferences and provided recovery average values from 84% to 120% with relative standard deviations which varied from 3.2 to 7.2%. PMID- 16362292 TI - Assembly of layer-by-layer films of heme proteins and single-walled carbon nanotubes: electrochemistry and electrocatalysis. AB - After being treated by mixed acids, single-walled carbon nanotubes (SWNTs) were shortened and had negatively charged groups on the surface. Positively charged hemoglobin or myoglobin at pH 5.0 was successfully assembled with SWNTs into layer-by-layer films on solid surfaces, designated as {SWNT/protein}n. While only those proteins in the first few bilayers closest to the electrode surface exhibited electroactivity, the {SWNT/protein}n films demonstrated a much higher fraction of electroactive proteins and better controllability in film construction compared with cast films of the proteins and carbon nanotubes. The proteins in the {SWNT/protein}n films retained their near-native structure at medium pH. The stable protein film electrode showed good electrocatalytic properties toward reduction of oxygen and hydrogen peroxide, demonstrating the potential application of the {SWNT/protein}n films as a new type of biosensor based on the direct electrochemistry of proteins without using mediators. PMID- 16362293 TI - Analysis of trace levels of pesticides in rainwater using SPME and GC-tandem mass spectrometry. AB - A multiresidue method using gas chromatography coupled to ion trap tandem mass spectrometry (GC-ITD-MS/MS) associated with solid phase microextraction (SPME) was developed for the analysis of 20 pesticides commonly used in the Alsace region in rainwater samples. Since the pesticides were expected to be present at very low concentrations and in complex matrices, the analytical method used was both highly selective and sensitive. Therefore, fibers coated with polyacrylate (PA), polydimethylsiloxane (PDMS) and polydimethylsiloxane-divinylbenzene (PDMS DVB) were tested, and the parameters affecting the precision and accuracy of the SPME method were investigated and optimized. These parameters include the type of fiber, the adsorption time, the effect of salt, and the extraction temperature. The PDMS fiber was the most polyvalent for the extractions of the different pesticides studied. Detection limits of between 5 and 500 ng L(-1), depending on the compounds under study (except for those which could not be analyzed: captan and mevinphos), were obtained with this analytical procedure. This method was applied to the analysis of rainwater samples collected simultaneously on a weekly basis at one rural and one urban site between March 2002 and July 2003. While some of the 20 pesticides analyzed were constantly detected (such as lindane and atrazine), a strong temporal variability was observed for some of the others (including alachlor, metolachlor, atrazine). PMID- 16362294 TI - A new continuous calibration method for inductively coupled plasma spectrometry. AB - A new calibration method was developed and applied to inductively coupled plasma atomic emission spectrometry. External calibration was performed as follows. A container was filled with a given volume of deionized (V(p)) water. Then a concentrated standard was introduced at a controlled rate (Q(e)) into the tank by means of a peristaltic pump. The resulting solution was stirred throughout the experiment. Simultaneously, the solution inside the tank was pumped from the vessel to the plasma at a given rate (Q(s)). The signal was continuously recorded. The variation of the concentration of the solution leaving the tank with time was determined by applying a basic equation of stirred tanks. The representation of the emission intensity versus the time and the further conversion of the time scale into a concentration scale gave rise to the calibration line. The best results in terms of linearity were achieved for V(p)=15 cm3, Q(e)=0.6-0.75 ml min(-1) and Q(s)=1-1.2 ml min(-1). Graphs with more than 40 standards were obtained within about 10 min. The results found were not statistically different from those afforded by the conventional calibration method. In addition, the new method was faster and supplied better linearity and precision than the conventional one. Another advantage of the stirred tank was that procedures such as dynamic calibration and standard additions could be easily and quickly applied, thus shortening the analysis time. A complete analysis following these procedures based on the measurement of 30 standards took about 5 min. Several synthetic as well as certified samples (i.e., bovine liver, mussel tissue and powdered milk) were analyzed with the stirred tank by applying four different calibration methodologies (i.e., external calibration, internal calibration, standard additions and a combination of internal standardization and standard additions), with the combination of internal standardization and standard additions being the method that provided the best results. The element concentrations obtained were not significantly different from the actual or certified values. PMID- 16362295 TI - Antitumor activity of erlotinib in combination with capecitabine in human tumor xenograft models. AB - PURPOSE: To examine the antitumor activity and tolerability of a combination comprising erlotinib and capecitabine in human colorectal, breast and epidermal cancer xenograft models. Further aims of the study were to examine the effects of single-agent erlotinib therapy on tumor growth, and on thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) levels, (enzymes which activate and deactivate capecitabine, respectively) in tumor tissue. METHODS: BALB/c nu/nu mice bearing LoVo and HT-29 (colon cancer), A-431 (vulval cancer), and KPL-4 and MAXF401 (breast cancer) human tumors were treated with erlotinib 100 mg/kg/day and/or capecitabine 359 or 90 mg/kg/day, by oral administration once daily for 14 days. RESULTS: The maximum tolerated dose (MTD) of erlotinib, formulated in carboxymethylcelluose/Tween 80, was identified as 125 mg/kg/day. Erlotinib at a dose of 100 mg/kg/day achieved significant tumor-growth inhibition in the, LoVo, KPL-4, and A-431 models. Some inhibition of MAXF401 tumor growth was observed, but was not significant. In the HT-29 model, erlotinib showed less marked but statistically significant antitumor activity. On day 15, mean tumor-growth inhibition in HT-29, LoVo, KPL-4, MAXF401, and A-431 models was 46, 74, 71, 20, and 85%, respectively. Evaluation of erlotinib/capecitabine combination therapy, at sub-optimal doses, in the three erlotinib-sensitive tumor models LoVo, KPL-4 and A-431, demonstrated at least additive activity with the combination compared with the single agents. In the A-431 and LoVo models, the combination of agents had greater antitumor activity than the single agent capecitabine alone at the MTD. Erlotinib in combination with capecitabine was not associated with significantly increased toxicity compared with single-agent therapy. Erlotinib 100 mg/kg/day induced significant upregulation of TP and DPD in the LoVo model, a significant upregulation of TP in the HT-29, MAXF401 and A-431 models, but had no obvious effect on TP and DPD levels in the KPL-4 model. In the A-431 model, selective upregulation of TP by erlotinib 100 mg/kg resulted in an increased TP:DPD ratio. In the LoVo model, immunohistochemistry revealed marked upregulation of TP (but not DPD by erlotinib). CONCLUSIONS: Erlotinib inhibits tumor growth in a range of human tumor xenograft models, including breast and colorectal cancer (CRC). Erlotinib and capecitabine demonstrated at least additive activity in LoVo, KPL-4 and A-431 tumor models. The antitumor activity of the combination was greater than that of capecitabine alone at the MTD. Erlotinib treatment did affect the TP in the CRC tumor models as confirmed immunohistochemically. The findings of this study support clinical evaluation of erlotinib, both as a single agent and in combination with capecitabine, for the treatment of CRC and breast cancer. PMID- 16362296 TI - Clindamycin-paclitaxel pharmacokinetic interaction in ovarian cancer patients. AB - INTRODUCTION: Plasma protein binding is an important factor for many drugs that can influence the tissue distribution and pharmacokinetics. alpha(1)-acid glycoprotein (AGP) is an acute-phase protein that can increase in plasma of patients with several pathological conditions including cancer. Studies performed in cultured cells indicate that paclitaxel cytotoxicity is reduced by adding AGP and the sensitivity to paclitaxel is restored by displacing its binding to AGP with clindamycin, resulting in an increased paclitaxel cell uptake. The purpose of this study was to evaluate whether clindamycin modifies paclitaxel pharmacokinetics also in cancer patients. PATIENTS AND METHODS: Sixteen patients with advanced ovarian cancer, previously treated with surgery and chemotherapy were enrolled in this study. A pharmacokinetic study of paclitaxel was performed in the first three cycles of the consolidation therapy (paclitaxel and carboplatin) in each patient. In these cycles paclitaxel was administered alone and with two different doses (600 and 1,200 mg) of concurrent clindamycin. The sequence of the three treatments was randomly assigned in each patient in order to avoid the same order of treatments. RESULTS: Paclitaxel pharmacokinetics were partly modified by the concurrent administration of clindamycin. C (max) and AUC(0-last) of paclitaxel were significantly higher when the drug was given alone than when it was coadministered with 1,200 mg clindamycin. Moreover, AGP concentrations seem to have a small but statistically significant influence on paclitaxel pharmacokinetic, since AUC(0-last) showed a positive significant correlation with AGP plasma concentration when paclitaxel was given alone. The linear relation was lost when paclitaxel was coadministered with 1,200 mg clindamycin. Toxicity was not influenced by the coadministration of clindamycin. CONCLUSION: The hypothesis that clindamycin could affect paclitaxel pharmacokinetics seems to be verified with this study. Nevertheless, changes induced by giving the combination of the two drugs are minimal and thus of questionable clinical relevance. PMID- 16362297 TI - Enforced expression of cytosolic 5'-nucleotidase I confers resistance to nucleoside analogues in vitro but systemic chemotherapy toxicity precludes in vivo selection. AB - PURPOSE: Retroviral transfer of cDNA sequences that confer drug resistance can be used to protect against chemotherapy-induced hematopoietic toxicity and for the selective expansion of gene-modified cells. To successfully expand genetically engineered cells in vivo, an appropriate balance must be achieved between systemic toxicity induced by the selecting agent and the expansion of modified cells. METHOD: In this study, we investigate retroviral transfer of cytosolic 5' nucleotidase I (cN-I) for protection and selection of gene-modified cells when treated with 2-chloro-2'-deoxyadenosine (2-CdA) and 5-fluorouracil (5-FU) alone and in combination. We also attempt to design a treatment strategy for the potential in vivo selection of cN-I-modified cells by administering 5-FU to mice prior to 2-CdA treatment. RESULTS: Our results show that cN-I can be transferred by recombinant retroviruses, and that enforced expression of cN-I protects murine fibroblast and hematopoietic progenitor cells from the cytotoxic effects of 2-CdA and/or 5-FU. Furthermore, we show that the combined administration of 5-FU and 2 CdA potentiates hematopoietic stem cell toxicity. However, the treatment also results in severe myelosuppression. CONCLUSION: These results show that while cN I provides both protective and selective benefits to gene-modified cells in vitro, selection requires a treatment strategy that is likely too toxic to consider cN-I as an in vivo selectable marker. PMID- 16362298 TI - Multidrug resistance proteins and folate supplementation: therapeutic implications for antifolates and other classes of drugs in cancer treatment. AB - Over the past decades, numerous reports have covered the crucial role of multidrug resistance (MDR) transporters in the efficacy of various chemotherapeutic drugs. Specific cell membrane-associated transporters mediate drug resistance by effluxing a wide spectrum of toxic agents. Although several excellent reviews have addressed general aspects of drug resistance, this current review aims to highlight implications for the efficacy of folate-based and other types of chemotherapeutic drugs. Folates are vitamins that are daily required for many biosynthetic processes. Folate supplementation in our diet may convey protective effects against several diseases, including cancers, but folate supplementation also makes up an essential part of several current cancer chemotherapeutic regimens. Traditionally, the folate leucovorin, for instance, is used to reduce antifolate toxicity in leukemia or to enhance the effect of the fluoropyrimidine 5-fluorouracil in some solid tumors. More recently, it has also been noted that folic acid has the ability to increase antitumor activity of several structurally unrelated regimens, such as alimta/pemetrexed and cisplatin. Moreover, studies from our laboratory demonstrated that folates could modulate the expression and activity of at least two members of the MDR transporters: MRP1/ABCC1, and the breast cancer resistance protein BCRP/ABCG2. Thus, folate supplementation may have differential effects on chemotherapy: (1) reduction of toxicity, (2) increase of antitumor activity, and (3) induction of MRP1 and BCRP associated cellular drug resistance. In this review the role of MDR proteins is discussed in further detail for each of these three items from the perspective to optimally exploit folate supplementation for enhanced chemotherapeutic efficacy of both antifolate-based chemotherapy and other classes of chemotherapeutic drugs. PMID- 16362299 TI - Phase I clinical trial of the farnesyltransferase inhibitor BMS-214662 administered as a weekly 24 h continuous intravenous infusion in patients with advanced solid tumors. AB - PURPOSE: BMS-214662 is a novel farnesyltransferase (FT) inhibitor that has shown promising suggestions of single agent activity in patients with advanced solid tumors when administered as a 1 h intravenous (i.v.) infusion every 3 weeks. The degree of FT inhibition in peripheral blood mononuclear cells (PBMCs) was greatest at the end of the infusion and rapidly reversed as the concentration of the drug in the plasma decayed. A second phase I trial of BMS-214662 administered as a weekly 24 h i.v. infusion was initiated to determine if the duration of maximum FT inhibition could be significantly extended by prolonging the infusion time and increasing the frequency of administration. PATIENTS AND METHODS: Infusion of BMS-214662 was prolonged from 2, 4, 8, 16, 24 h in single patient cohorts and repeated weekly for 3 out of 4 weeks. The initial dose was 56 mg/m(2). When the infusion duration reached 24 h, the dose was escalated at a constant multiples of 1.4 in single patient cohorts until the occurrence of toxicity greater than grade 1, upon which groups of at least three patients were evaluated at each dose level. The plasma pharmacokinetics and FT inhibition in PBMCs were measured in all patients at the prospective maximum tolerated dose. RESULTS: Nineteen patients participated in the study (11 males/8 females) and the weekly dose was increased to a maximum of 300 mg/m(2) given as a 24 h i.v. infusion. Drug-related toxicity greater than grade 1 first occurred at 300 mg/m(2), with two patients experiencing dose-limiting toxicity. One patient developed a grade 3 hyponatremia and another developed reversible grade 3 diarrhea, grade 2 renal toxicity, and grade 3 transaminitis. A 275 mg/m(2) dose was then evaluated, where one of the three patients treated experienced reversible grade 4 renal toxicity and grade 3 diarrhea. In view of the identical renal toxicity at 275 mg/m(2) in another study and limited drug availability, there was no further accrual to this dose level and the study was closed. No evidence of antitumor activity was observed. The plasma pharmacokinetics of BMS 214662 was linear with high interpatient variability. In the three patients evaluated at the 275 mg/m(2) dose level, the maximum inhibition of FT activity in PBMCs was 47+/-23% of the baseline. CONCLUSION: Administering BMS-214662 as a weekly 24 h continuous i.v. infusion permitted a considerably greater dose intensity to be delivered as compared to a single 1 h infusion given once every 3 weeks. The more prolonged infusion schedule resulted in a much lower degree of maximum FT inhibition in PBMCs than achieved with the 1 h infusion, although the duration of enzyme inhibition was longer, consistent with the lower peak plasma concentration of the drug provided by comparably tolerated doses when given as a 24 h infusion. Similarly, delivering the drug with increased dose intensity permitted by this weekly administration schedule did not appear to enhance its therapeutic benefit, at least in this phase I trial. Continued development of BMS 214662 may depend upon the potential for using it in combination with other anticancer drugs. PMID- 16362300 TI - Complete sequence and organization of the cucumber (Cucumis sativus L. cv. Baekmibaekdadagi) chloroplast genome. AB - The nucleotide sequence of the cucumber (Cucumis sativus L. cv. Baekmibaekdadagi) chloroplast genome was completed. The circular double-stranded DNA, consisting of 155,527 bp, contained a pair of inverted repeat regions (IRa and IRb) of 25,187 bp each, which were separated by small and large single copy regions of 86,879 and 18,274 bp, respectively. The presence and relative positions of 113 genes (76 peptide-encoding genes, 30 tRNA genes, four rRNA genes, and three conserved open reading frames) were identified. The major portion (55.76%) of the C. sativus chloroplast genome consisted of gene-coding regions (49.13% protein coding and 6.63% RNA regions; 27.81% LSC, 9.46% SSC and 18.49% IR regions), while intergenic spacers (including 20 introns) made up 44.24%. The overall G-C content of C. sativus chloroplast genome was 36.95%. Sixteen genes contained one intron, while two genes had two introns. The expansion/contraction manner of IR at IRb/LSC and IR/SSC border in Cucumis was similar to that of Lotus and Arabidopsis, and the manner at IRa/LSC was similar to Lotus and Nicotiana. In total, 56 simple sequence repeats (more than 10 bases) were identified in the C. sativus chloroplast genome. PMID- 16362301 TI - Agrobacterium-mediated genetic transformation of the desiccation tolerant resurrection plant Ramonda myconi (L.) Rchb. AB - In this paper we describe the first procedure for Agrobacterium tumefaciens mediated genetic transformation of the desiccation tolerant plant Ramonda myconi (L.) Rchb. Previously, we reported the establishment of a reliable and effective tissue culture system based on the integrated optimisation of antioxidant and growth regulator composition and the stabilisation of the pH of the culture media by means of a potassium phosphate buffer. This efficient plant regeneration via callus phase provided a basis for the optimisation of the genetic transformation in R. myconi. For gene delivery, both a standard (method A) and a modified protocol (method B) have been applied. Since the latter has previously resulted in successful transformation of another resurrection plant, Craterostigma plantagineum, an identical protocol was utilized in transformation of R. myconi, as this method may prove general for dicotyledonous resurrection plants. On this basis, physical and biochemical key variables in transformation were evaluated such as mechanical microwounding of plant explants and in vitro preinduction of vir genes. While the physical enhancement of bacterial penetration was proved to be essential for successful genetic transformation of R. myconi, an additional two-fold increase in the transformation frequency was obtained when the above physical and biochemical treatments were applied in combination. All R0 and R1 transgenic plants were fertile, and no morphological abnormalities were observed on the whole-plant level. PMID- 16362302 TI - Transformation of a muskmelon 'Galia' hybrid parental line (Cucumis melo L. var. reticulatus Ser.) with an antisense ACC oxidase gene. AB - 'Galia' muskmelon (Cucumis melo L. var. reticulatus Ser.) has been recalcitrant to transformation by Agrobacterium tumefaciens. Transformation of the 'Galia' male parental line, 'Krymka', with an ACC oxidase (CMACO-1) gene in antisense orientation is described herein. Explants were transformed using A. tumefaciens strain ABI, which contained a vector pCmACO1-AS plasmid, bearing an antisense gene of CMACO-1 and the CP4 syn gene (glyphosate-tolerance). Both CMACO-1 and CP4 syn genes were assessed by a polymerase chain reaction method. Flow cytometry analysis was performed to determine plant ploidy level of primary transformants. Two completely diploid independent transgenic plants were obtained. Southern blot and segregation analysis in the T1 generation determined that each independent transgenic line had one single insertion of the transgene. These transgenic muskmelon male parental lines have potential for use in the production of 'Galia' F1 hybrids with improved shelf life. PMID- 16362303 TI - Changes in gene expression in maize kernel in response to water and salt stress. AB - Increasing pressure on limited water resources for agriculture, together with the global temperature increase, highlight the importance of breeding for drought tolerant cultivars. A better understanding of the molecular nature of drought stress can be expected through the use of genomics approaches. Here, a macroarray of approximately 2500 maize cDNAs was used for determining transcript changes during water- and salt-stress treatments of developing kernels at 15 days after pollination. Normalization of relative transcript abundances was carried out using a human nebulin control sequence. The proportions of transcripts that changed significantly in abundance upon treatment (>2-fold compared to the control) were determined; 1.5% of the sequences examined were up-regulated by high salinity and 1% by water stress. Both stresses induced 0.8% of the sequences. These include genes involved in various stress responses: abiotic, wounding and pathogen attack (abscisic acid response binding factor, glycine and proline-rich proteins, pathogenesis-related proteins, etc.). The proportion of down-regulated genes was higher than that for up-regulated genes for water stress (3.2%) and lower for salt stress (0.7%), although only eight genes, predominantly involved in energy generation, were down-regulated in both stress conditions. Co expression of genes of unknown function under defined conditions may help in elucidating their roles in coordinating stress responses. PMID- 16362304 TI - Auditory lateralization in bushcrickets: a new dichotic paradigm. AB - Pair formation in the bushcricket Gampsocleis gratiosa is achieved through acoustic signalling by the male and phonotactic approaches of the female towards the calling song. On a walking belt in the free sound field, females tracked the position of the speaker broadcasting the male calling song with a remarkable precision, deviating by no more than 10 cm in either direction from the ideal course. Starting with stimulus angles of 6-10 degrees the females significantly turned to the correct side, and with stimulus angles greater than 25 degrees no incorrect turns were made. Using neurophysiological data on the directionality of the ear we calculated that with such stimulus angles the available binaural intensity difference is in the order of 1-2 dB. We developed a dichotic ear stimulation device for freely moving females with a cross-talk barrier of about 50 dB, which allowed to precisely apply small binaural intensity differences. In such a dichotic stimulation paradigm, females on average turned to the tronger stimulated side starting with a 1 dB difference between both ears. The significance of such a reliable lateralization behaviour with small interaural intensity differences for phonotactic behaviour under natural conditions is discussed. PMID- 16362305 TI - Common motor mechanisms support body load in serially homologous legs of cockroaches in posture and walking. AB - We studied the mechanisms underlying support of body load in posture and walking in serially homologous legs of cockroaches. Activities of the trochanteral extensor muscle in the front or middle legs were recorded neurographically while animals were videotaped. Body load was increased via magnets attached to the thorax and varied through a coil below the substrate. In posture, tonic firing of the slow trochanteral extensor motoneuron (Ds) in each leg was strongly modulated by changing body load. Rapid load increases produced decreases in body height and sharp increments in extensor firing. The peak of extensor activity more closely approximated the maximum velocity of body displacement than the body position. In walking, extensor bursts in front and middle legs were initiated during swing and continued into the stance phase. Moderate tonic increases in body load elicited similar, specific, phase dependent changes in both legs: extensor firing was not altered in swing but was higher after foot placement in stance. These motor adjustments to load are not anticipatory but apparently depend upon sensory feedback. These data are consistent with previous findings in the hind legs and support the idea that body load is countered by common motor mechanisms in serially homologous legs. PMID- 16362306 TI - HIF-1alpha and iNOS levels in crucian carp gills during hypoxia-induced transformation. AB - Hypoxia inducible factor 1 alpha (HIF-1alpha) initiates expression of a wide variety of genes, some of which are involved in apoptosis and cell cycle arrest. We have previously shown that crucian carp increases its respiratory surface area 7.5-fold in response to hypoxia. This change is due to apoptosis and cell cycle arrest in specific parts of its gills. Here we have characterized crucian carp HIF-1alpha, and measured mRNA, protein and DNA binding levels during hypoxia exposure in crucian carp gills. We have also measured an HIF-1alpha-induced gene, the inducible nitric oxide synthase (iNOS), which has the ability to initiate apoptosis and cell cycle arrest. Crucian carp HIF-1alpha was found to have all critical domains known to be important for function. Comparison of the peptide sequence with other species indicated high similarity with other cyprinid fish, but a pronounced variation compared to the salmonid, rainbow trout. Further, we found HIF-1alpha protein to be stabilized during hypoxia. Further, HIF-1alpha was often present in normoxia, and showed marked individual weight-dependent variation. We found no alteration of iNOS mRNA levels during hypoxia exposure. These findings suggest HIF-1alpha involvement in hypoxia-induced change of respiratory surface area in crucian carp gills. However, its activity does not seem to be mediated through iNOS. PMID- 16362307 TI - Direct influence of serotonin on the larval heart of Drosophila melanogaster. AB - The heart rate (HR) of larval Drosophila is established to be modulated by various neuromodulators. Serotonin (5-HT) showed dose-dependent responses in direct application within semi-intact preparations. At 1 nM, HR decreased by 20% while it increased at 10 nM (10%) and 100 nM (30%). The effects plateaued at 100 nM. The action of 5-HT on the heart was examined with an intact Central Nervous System (CNS) and an ablated CNS. The heart and aorta of dorsal vessel pulsate at different rates at rest and during exposure to 5-HT. Splitting the heart and aorta resulted in a dramatic reduction in pulse rate of both the segments and the addition of 5-HT did not produce regional differences. The split aorta and heart showed a high degree of sensitivity to sham changes of saline but no significant effect to 5-HT. Larvae-fed 5-HT (1 mM) did not show any significant change in HR. Since 3,4-methylenedioxymethamphetamine (MDMA) is known to act as a weak agonist on 5-HT receptors in vertebrates, we tested an exogenous application; however, no significant effect was observed to dosage ranging from 1 nM to 100 microM in larvae with and without an intact CNS. In summary, direct application of 5-HT to the larval heart had significant effects in a dose-dependent manner while MDMA had no effect. PMID- 16362308 TI - Metabolic rate of Arctic charr eggs depends on their parentage. AB - The metabolic rate (specific heat output) of individual eyed-stage eggs of Arctic charr Salvelinus alpinus (Linnaeus, 1758) originating from different families was measured with direct microcalorimetry. Metabolic rates varied between 2.3-7.9 microW ind(-1) and 0.06-0.22 microW mg(-1). Absolute heat output was unrelated to egg size, but size-scaled or specific heat output was negatively correlated with egg size, measured as diameter, dry mass or fresh mass. Metabolic rates varied significantly between families, suggesting that genetic and/or maternal effects affect embryonic metabolism in Arctic charr. Heat output increased almost linearly from 3.4 to 16.7 microW ind(-1) (0.09-0.67 microW mg(-1)) during the embryonic development. Although the metabolic rate varied between the families and egg metabolic rate increased during development, there was an unexpected disconnect between metabolic rate and hatching time. PMID- 16362309 TI - Complicated pneumonias with empyema and/or pneumatocele in children. AB - To investigate the incidence, procedure type, characteristics of pleural fluid and pneumatoceles, and evolution of pneumonia complicated with empyema and/or pneumatoceles. Review of 394 pediatric pneumonia in patients at Sao Paulo State University Hospital during 2 years. We studied those with complications such as pleural effusion and pneumatocele. There were 121 (30.71%) with complications such as pleural effusion and pneumatocele; these were significantly higher in infants. One hundred and six children were needle aspirated, of these 78 underwent drainage, and 15 observation only. From the drained, seven needed thoracotomy or pleurostomy. Fluid was purulent in 50%, and pneumatoceles were seen in 33 cases (8.3%) with spontaneous involution in 28 (85%). Pleural fluid culture was negative in 51% cases; in positive cultures, Streptococcus pneumoniae was the most common agent. Complicated pneumonia incidence was higher in the second year of life and more than 70% occurred before 4 years of age. Closed thoracic drainage was effective in over 90%. Large effusions and mediastinal deviations were submitted to more aggressive procedures. Pneumatoceles predominated in the under 3s and were generally evident in the first chest X-ray. Most cases had spontaneous pneumatocele involution, and in almost half the cases were still present at drain tube removal. PMID- 16362310 TI - The impact of Down's syndrome on the immediate and long-term outcomes of children with Hirschsprung's disease. AB - Hirschsprung's disease (HD) in Down's syndrome (DS) patients is stated to have a worse outcome than HD alone. In our study we reviewed the immediate and long-term outcomes of these children and questioned whether DS should influence the operative management. Data were collected on all children with HD (including total colonic aganglionosis), between January 1990 and December 2000. They were divided into two groups based on the presence or absence of Trisomy 21 and compared retrospectively. In this time period we treated 173 children with HD; 17 of these had DS. Both the groups were comparable in their mean gestational age, birth weight and presentation except that the DS group had a significantly higher overall incidence of pre and/or postoperative enterocolitis. A tota1 of 164 children underwent a Swenson pull-through and 9 had a Soave's procedure. Follow up ranged from 1 to 10 years. Continence assessed using the Wingspread scoring system in children over the age of 4 years showed no significant difference. Although children with both HD and DS are predisposed to complications and required a more cautious management, long-term outcome in terms of continence was not significantly worse than in HD alone. Thus the co-existence of DS should not influence the decision to offer these children and their parents the choice of definitive repair. PMID- 16362311 TI - A study of abnormal cervical cytology in pregnant women. AB - During the 4 year routine study of smears in 2,919 pregnant women, 33 cases of abnormals of the uterine cervix were detected (1.13%). The patients were followed with uterine cervical cytology and colposcopy and in case of need, sometimes punch biopsies were performed. The results of the cytologies, 33 cases with abnormalities were detected. There were 26 cases classified as class IIIa, 7 cases were class IIIb. All the cases underwent colposcopy. For the 17 cases that showed lesions by colposcopy, and punch biopsies were performed. The results of histologic examination were wide variety, five chronic cervicitis, one condyloma, one mild dysplasia, three moderate dysplasia, three severe dysplasia, three carcinoma in situ, and one microinvasive carcinoma. Only two cases were treated during pregnancy; one with condyloma underwent Laser vaporization and another with microinvasive carcinoma underwent LEEP conization. Other cases were given conservative treatment during pregnancy. Excluding one case for persistence smear class IIIa of histology condyloma, all the cases showed regression of dysplasia and carcinoma in situ with treatment after delivery. We conclude that lesions up to carcinoma in situ do not require intervention during pregnancy but microinvasive carcinoma is suspected, diagnostic LEEP conization is necessary, even during pregnancy. PMID- 16362312 TI - Recurrent early pregnancy loss and endothelial nitric oxide synthase gene polymorphisms. AB - OBJECTIVE(S): Studies on the relation between endothelial nitric oxide synthase (eNOS) activity in implantation and maintenance of pregnancy highlights the importance of eNOS gene polymorphisms in recurrent early pregnancy loss (REPL). We investigated the relationship between idiopathic REPL and polymorphisms in eNOS among South Indian women. METHODS: A case-control study comprising 145 females with REPL and 99 control females. The polymorphisms studied include a 27 bp intron 4 repeat, Glu298Asp variation of exon 7 and a novel 140 A --> G polymorphism in intron 6. A polymerase chain reaction-based di-deoxy dye terminator sequencing method was used for genotyping. RESULTS: A novel A --> G polymorphism was identified in intron 6. The more frequent b allele of intron 4 repeat was present at a frequency of 0.84 in cases as compared to 0.86 in controls (O.R 1.17); the G allele of exon 7 coding for the wild-type glutamate containing isoform was present at a frequency of 0.79 in cases and 0.83 in controls (O.R 1.30, CI 0.6-2.8). The intron 6 variant A allele was present at a frequency of 0.58 in cases and 0.45 in controls (O.R 0.59, CI 0.33-1.08). Overall, the polymorphism in intron 6, in homozygous condition, exhibited a significant association to the risk of REPL (O.R 0.43, CI 0.21-0.89), P: 0.021). CONCLUSIONS: The present study identifies and validates a novel polymorphism in the eNOS gene which was found associated with the risk of REPL. PMID- 16362313 TI - Lack of association between insulin-like growth factor I receptor G(+3174)A polymorphism and retinopathy of prematurity. AB - BACKGROUND: Retinopathy of prematurity (ROP) is a potentially blinding eye disorder that affects premature infants. Insulin-like growth factor I (IGF-I) has been established as being necessary for vascular growth in the neonatal retina. Low IGF-I levels during the early postnatal days have been found to be predictive for ROP. The effects of IGF-I are mediated through IGF-I receptors (IGF-IR), which, in turn, suppress IGF-I production. The G(+3174)A polymorphism of the IGF IR gene has been shown to be associated with low IGF-I levels. We tested the association of this IGF-IR polymorphism with ROP. METHODS: We enrolled in the ROP group those infants (n=108) who had been treated with laser or cryo therapy due to ROP stage 2+ or 3 (n=91) or had a ROP stage 4 or 5 (n=17) (ROP group). The median gestational age of these infants was 28 weeks (range 24-35 weeks) and birth weight was 970 g (range 630 to 2,000 g). The distribution of IGFR-1 G(+3174)A genotype in the ROP group was compared to that in 120 gestational age matched infants with ROP stage 1 or 2 not requiring intervention [gestational age 30 (range 24-37) weeks, birth weight 1,235 (640-1,960) g] (LBW group) and 164 term newborns [gestational age 39 (range 35-42) weeks, birth weight 3,450 (2,500 4,350) g] (term group). Genotyping was done using PCR-RFLP methods. RESULT: The prevalence of IGF-IR G(+3174)A polymorphism was the same in the ROP group, the LBW group and the term group, showing no association between this single nucleotide polymorphism (SNP) and ROP. CONCLUSION: Our results do not support the hypothesis that the carrier state of IGF-IR G(+3174)A polymorphism has an impact on the risk of ROP in infants. A possible cause for the lack of association is that the rapid nutritional and metabolic changes during postnatal life have a greater effect on IGF-I levels than this SNP does. PMID- 16362314 TI - Allergic reaction to hyaluronidase: a rare cause of orbital inflammation after cataract surgery. AB - PURPOSE: The aim of this study was to present a series of patients with acute orbital inflammatory symptoms secondary to peribulbar hyaluronidase allergy and to discuss the diagnostic and management modalities. METHODS: Review of clinical records of all patients with acute orbital inflammatory symptoms after uneventful cataract surgery, seen at two ophthalmology departments. The main outcome measures included clinical signs and symptoms, imaging findings, post-operative course, skin allergy testing, and final outcome. RESULTS: There were five patients (four women, one man) with a mean age of 78+/-7.0 years. Signs and symptoms appeared 12-72 h after cataract surgery and included axial proptosis, periorbital erythema with swelling, and extraocular muscle (EOM) function restriction (5/5), periorbital pain or itchiness (3/5), and conjunctival chemosis (4/5). Computerized tomography showed increased orbital fat haziness and enlargement of EOM. Treatment with a combination of oral antibiotics and steroids or antihistamines resulted in resolution of signs and symptoms after 3-5 days. Intradermal and/or skin prick allergy tests were positive for hyaluronidase and negative for all other perioperative medications used. CONCLUSION: Although uncommon, allergy to peribulbar hyaluronidase injected during cataract surgery should be considered in the differential diagnosis of patients who present with acute post-operative orbital signs and symptoms. PMID- 16362315 TI - Eyelid splitting with follicular extirpation using a monopolar cautery for the treatment of trichiasis and distichiasis. AB - PURPOSE: To assess the efficacy of eyelid splitting coupled with follicular extirpation via monopolar cautery in the treatment of trichiasis and distichiasis. METHODS: Between March 2000 and October 2003, this surgery was performed on 52 eyelids from 45 patients, with a mean follow-up period of 14.3 months (ranging from 12.3 to 17.6 months) . Under local anesthesia and a surgical microscope, a chalazion clamp is positioned on the eyelid, and a no. 11 scalpel is employed to make an incision located immediately anterior and parallel to the abnormal eyelash line. Each of the abnormal hair follicles is then removed via cautery with a monopolar needle. No sutures are involved in this procedure. RESULTS: Forty-four eyelids of 40 patients (84.6%) were successfully treated without recurrence or any residual symptoms. Repeat surgery was then performed on eight eyelids, with successful results in six cases. The cumulative success rate for this procedure was 96.1%. No complications were noted in the treated area, including lid deformities, granuloma formation, infections, and others. CONCLUSIONS: Eyelid splitting coupled with follicular extirpation via cautery with a monopolar needle, a procedure which requires no sutures, constitutes a simple and effective method for the treatment of both trichiasis and distichiasis and is associated with favorable functional and cosmetic results. PMID- 16362316 TI - An objective technique to measure the depth-of-focus in free space. AB - BACKGROUND: Determining the depth-of-focus (DOF) objectively in free space is important, as this provides information regarding the range of clear vision under more natural viewing conditions in which target blur, size, and proximal information, as well as other monocular depth cues, are present. METHODS: The DOF was assessed objectively in free space in young adults [n=20, ages 24.9+/ 3.1years: myopes (n=14), emmetropes (n=5), hyperope (n=1)] by monitoring accommodation continuously using the commercially available Power Refractor II (PR II) under monocular viewing conditions. A high-contrast target ( approximately 83%) subtending a mean visual angle of 2.3 deg was placed on an optical bench at 25 cm (4 D) along the line-of-sight of the viewing right eye and was displaced slowly ( approximately 0.1-0.15 D/s) proximally and distally. This target provided blur, size, and proximal information, and other depth cues, as normally found in free space. The first consistent change in steady-state accommodative baseline level reflected one edge of the DOF. The mean dioptric difference between these distal and proximal endpoints was averaged, and this represented the total objective DOF. RESULTS: The group mean total objective DOF (n=20) was +/-0.61+/-0.09 D, with a range from +/-0.46 D to +/-0.81 D. Repeatability was excellent. CONCLUSION: A technique was developed and tested to measure the DOF in free space objectively. Further development of this technique will allow the assessment of blur perception under more complex natural viewing conditions simulating the everyday environment. PMID- 16362318 TI - Demarcation laser therapy in the management of macular-sparing persistent subretinal fluid after scleral buckling procedures. AB - BACKGROUND: Persistent or recurrent macular-sparing subretinal fluid (SRF) can sometimes occur following scleral buckling procedures. Observation and reoperation have been used in the management of such cases. Demarcation laser therapy (DLT) has been used to treat macular-sparing retinal detachments in the context of cytomegalovirus retinitis and as primary treatment for selected rhegmatogenous retinal detachments. There are, however, scarce data in the literature regarding its use following primary scleral buckling procedures. The current study explores the use of DLT under the latter circumstances. METHODS: The medical records of all consecutive patients with persistent SRF sparing the macula following primary rhegmatogenous retinal detachment repair using a scleral buckling procedure were retrospectively reviewed. Only those patients in whom the breaks were localised to the area of indentation and, thus, seemed to be well supported by the buckle were included. Demographics, clinical characteristics of the retinal detachment prior to scleral buckling, extension of the residual SRF observed postoperatively, details of the laser procedure, anatomical and functional outcomes and complications were evaluated. RESULTS: Seven patients, all females, with a mean age of 47.9 years (range: 20-81) were included in the study. The retinal detachments were superior (n=3), inferior (n=3) and subtotal, affecting both superior and inferior retina (n=1). Scleral buckling procedures were used to treat the retinal detachments in all cases. Following demarcation laser therapy, the area of SRF remained stable in two patients, and flattened in four. In one patient, extension of SRF occurred requiring further surgery. CONCLUSIONS: Demarcation laser therapy appears to be a reasonable option in the management of patients with persistent or recurrent SRF sparing the macula following scleral buckling surgery. PMID- 16362317 TI - Nosocomial postoperative endophthalmitis: a 14-year review. AB - BACKGROUND: The objective of this study was to evaluate the incidence, predisposing surgery, management and final visual outcome of postoperative endophthalmitis over a 14-year period at one institute. METHODS: This retrospective study reviewed all intraocular operations performed between 1 January 1991 and 31 August 2004 at Chang Gung Memorial Hospital, Kaohsiung Medical Center, to determine the incidence of nosocomial postoperative endophthalmitis and the characteristics of patients who developed this condition. RESULTS: The overall incidence of postoperative endophthalmitis after intraocular surgery was 0.19% (56 out of 30,219). Postoperative endophthalmitis developed in 56 eyes in 56 patients during the study period. The condition developed after cataract surgery in 46 eyes, after penetrating keratoplasty in 6 eyes, after filtering surgery in 2 eyes, after secondary intraocular lens implant in 1 eye, and after vitrectomy in 1 eye. Postoperative endophthalmitis was culture-positive in 31 cases (55%). The most frequent organism isolated was coagulase-negative Staphylococcus. Factors associated with better visual acuity outcomes included low virulence of isolated pathogen, initial visual acuity of counting fingers or better, and history of cataract surgery compared with other intraocular surgery. CONCLUSION: The overall incidence of endophthalmitis after intraocular surgery was 0.19%. The results of this 14-year review from a local medical center may serve as a source of comparison for other centers and future studies. PMID- 16362319 TI - Changes in astigmatism in children with congenital nystagmus. AB - BACKGROUND: Astigmatism is commonly reported in children with and without nystagmus. In children less than 4 years of age the astigmatism changes from against the rule (ATR) to with the rule (WTR) astigmatism in children without nystagmus. However, little is known about children with congenital nystagmus. We compared astigmatism in children with congenital nystagmus below 4 years and above 4 years of age. MATERIAL AND METHODS: Three hundred and fifty-six eyes in 178 children who satisfied the study criteria were included. The children were divided into those below 4 years of age (n of eyes = 192) and those above (n of eyes = 164). Cycloplegic refraction (with manual retinoscopy) carried out at presentation and at the last follow-up were recorded and compared. Cycloplegia was achieved using cyclopentolate eye drops. Outcome of eyes with no astigmatism at initial presentation was compared with the final refraction in both the age groups. The visual acuity at the first presentation was compared with the visual acuity at the last presentation. RESULTS: Average follow-up duration was 3.36+/ 1.59 years. On presentation, 176 (49.44%) eyes did not have any astigmatism. We found that 26 (25.2%) out of the 103 children below 4 years of age who did not have any astigmatism on presentation developed WTR astigmatism after a mean follow-up of 3.5+/-1.5 years. (p= 0.042). In children >or=4 years of age only 7 (9.6%) children out of 73 developed WTR after a mean follow-up of 3.4+/-1.4 years. The visual acuity change was not found to be significant in the two groups. The majority (90.3%) improved or had same visual acuity in group A and 88.9% improved or had the same visual acuity in group B at the last follow-up (p= 0.77). DISCUSSION: Our incidence of WTR astigmatism in children with congenital nystagmus is similar to those previously reported series. Our data suggest that there is a significant chance that children under 4 years with congenital nystagmus may develop WTR astigmatism compared with children above 4 years of age. CONCLUSION: With the rule (WTR) astigmatism is common in children with nystagmus. Children under 4 years of age presenting with no astigmatism may acquire WTR astigmatism as they grow. The amount of astigmatism increases with age in children with nystagmus. Visual acuity, however, remains stable as the age advances. PMID- 16362320 TI - Ca2+ -activated K+ channels of the BK-type in the mouse brain. AB - An antibody against the 442 carboxy-terminal amino acids of the BK channel alpha subunit detects high immunoreactivity within the telencephalon in cerebral cortices, olfactory bulb, basal ganglia and hippocampus, while lower levels are found in basal forebrain regions and amygdala. Within the diencephalon, high density was found in nuclei of the ventral and dorsal thalamus and the medial habenular nucleus, and low density in the hypothalamus. The fasciculus retroflexus and its termination in the mesencephalic interpeduncular nucleus are prominently stained. Other mesencephalic expression sites are periaquaeductal gray and raphe nuclei. In the rhombencephalon, BK channels are enriched in the cerebellar cortex and in the locus coeruleus. Strong immunoreactivity is also contained in the vestibular nuclei, but not in cranial nerves and their intramedullary course of their roots. On the cellular level, BK channels show pre and postsynaptic localizations, i.e., in somata, dendrites, axons and synaptic terminals. PMID- 16362321 TI - Sperm DNA damage correlates with polycyclic aromatic hydrocarbons biomarker in coke-oven workers. AB - OBJECTIVES: The aim was to determine whether occupational exposure to polycyclic aromatic hydrocarbons (PAHs) in men has adverse effect on semen quality. METHODS: Forty-eight coke-oven workers, including 16 topside-oven (TO) workers and 32 sideoven (SO) workers, were studied. Ambient PAHs exposure, urinary 1 hydroxypyrene (1-OHP) levels, and parameters of semen quality were determined. RESULTS: TO workers had significantly higher ambient PAHs exposure (3,436.1+/ 3,411.0 vs. 1,123.1+/-1,829.3 ng/m3, P < 0.01), urinary 1-OHP levels (207.8+/ 176.4 vs. 54.0+/-44.8 microg/g, P < 0.001), frequency of oligospermia (18.8 vs. 0%, P < 0.05), and morphological abnormality in sperm (32.3 vs. 14.6%, P < 0.01) than SO workers. Among cigarette smokers, TO workers had significantly higher rates of DNA denaturation in spermatozoa (alphaT) and percentage of sperm with increased DNA denaturation (COMP alphaT) levels than SO workers (246.2+/-49.5 vs. 198.1+/-30.3 for alphaT; 34.8+/-14.4 vs. 19.3+/-13.9% for COMP alphaT, P < 0.05). There was a positive correlation between urinary 1-OHP and alphaT, COMP alphaT, and abnormal sperm morphology and a tenfold increase in urinary 1-OHP associated with a 2.35-fold increase in alphaT, as well as a 1.07-fold increase in percentage of sperm with abnormal morphology. CONCLUSIONS: A potential risk of sperm dysfunction should be considered for workers occupationally exposed to high levels of PAHs. Cigarette smoking may aggravate this risk. Urinary 1-OHP can be used as a biomarker predicting sperm dysfunction. PMID- 16362322 TI - Ambient monitoring and biomonitoring of workers exposed to N-methyl-2-pyrrolidone in an industrial facility. AB - OBJECTIVES: The exposure of seven workers and three on-site study examiners to N methyl-2-pyrrolidone (NMP) was studied in an adhesive bonding compound and glue production facility. METHODS: Airborne NMP was analysed by personal and stationary sampling on activated charcoal tubes. NMP and its main metabolites, 5 hydroxy-N-methyl-2-pyrrolidone (5-HNMP) and 2-hydroxy-N-methylsuccinimide (2 HMSI), were analysed in pre-shift and post-shift spot urine samples by gas chromatography-mass spectrometry. The workers were examined with respect to irritation of the eyes, the mucous membranes and the skin, and health complaints before and after the work-shift were recorded. RESULTS: The time-weighted average concentration of NMP in most work areas varied between 0.2 and 3.0 mg/m3. During the manual cleaning of stirring vessels, valves and tools, 8-h TWA exposures of up to 15.5 mg/m3 and single peak exposures of up to 85 mg/m3) were observed. NMP and its metabolites were detected in two pre-shift urine specimens. NMP and 5 HNMP concentrations in post-shift urine samples of five workers and three on-site study examiners were below 125 microg/g creatinine and 15 mg/g creatinine, respectively, while two vessel-cleaning workers showed significantly higher urinary NMP concentrations of 472 and 711 microg/g creatinine and 5-HNMP concentrations of 33.5 and 124 mg/g creatinine. 2-HMSI was detectable in four post-shift samples (range: 1.6-14.7 mg/g creatinine). The vessel cleaner with the highest NMP exposure reported irritation of the eyes, the upper respiratory tract and headaches. CONCLUSIONS: The results of this study indicate a relatively low overall exposure to NMP in the facility. An increased uptake of NMP occurred only during extensive manual vessel cleaning. Health complaints associated with NMP exposure were recorded in one case and might be related to an excessive dermal exposure due to infrequent and inadequate use of personal protective equipment. PMID- 16362323 TI - Effort-reward imbalance, sleep disturbances and fatigue. AB - OBJECTIVES: The objective of this study was to investigate the validity of the effort-reward imbalance (ERI) model in relation to disturbed sleep and fatigue. METHODS: The study population derived from a subset of the WOLF (WOrk, Lipids, Fibrinogen) cohort study of cardiovascular risk in a working population who replied to the ERI-questionnaire comprising 789 men and 214 women. Cox regression analysis was used to calculate the prevalence ratio (PR) for sleep disorders and fatigue in relation to the components of ERI. RESULTS: As sleep disturbances and fatigue, based on literature, were defined to be represented by the uppermost quintile, 14% of the men and 23% of the women were affected by sleep disturbances while 14 and 26%, respectively, were affected by fatigue. Higher levels of exposure for the ERI components were associated with increased prevalence of sleep disturbances and fatigue. For men, the strongest association was seen between high overcommitment and fatigue (PR 5.77, 95% confidence interval 2.89 11.5). For women, high effort and sleep disturbances (PR 4.04, CI 1.53-10.7), high effort/reward ratio and sleep disturbances (PR 4.13, CI 1.62-10.5), and between low reward and fatigue (PR 4.36, CI 1.79-10.6) yielded the most obvious associations. CONCLUSIONS: The present study adds sleep disturbances and fatigue to the list of adverse consequences of effort-reward imbalance. PMID- 16362324 TI - Changes in chloroplast DNA during development in tobacco, Medicago truncatula, pea, and maize. AB - We examined the DNA from chloroplasts obtained from young and fully expanded leaves of tobacco (Nicotiana tabacum L.), Medicago truncatula, pea (Pisum sativum L.), and maize (Zea mays L.). The changes in plastid DNA content and structure were monitored by four independent methods: 4',6-diamidino-2-phenylindole (DAPI) staining with intact chloroplasts, in situ DAPI staining of cytological sections, ethidium bromide staining at the single-molecule level after exhaustive deproteinization of lysed chloroplasts, and pulsed-field gel electrophoresis. During leaf development, we found a decline of chloroplast DNA (cpDNA) in all four plants. For tobacco, for which plants can readily be regenerated from somatic cells, cpDNA persisted longer than in the other three plants. We also found a striking progression from complex multigenomic DNA molecules to simple subgenomic molecules during plastid development. Although the decrease in molecular size and complexity paralleled the decrease in DNA content per plastid, 6% of the chloroplasts in a fully expanded tobacco leaf still contained DNA in complex branched structure, whereas no such complex structures were found in mature leaves for the hard-to-regenerate maize. PMID- 16362325 TI - Zinc compartmentation in root, transport into xylem, and absorption into leaf cells in the hyperaccumulating species of Sedum alfredii Hance. AB - Sedum alfredii Hance can accumulate Zn in shoots over 2%. Leaf and stem Zn concentrations of the hyperaccumulating ecotype (HE) were 24- and 28-fold higher, respectively, than those of the nonhyperaccumulating ecotype (NHE), whereas 1.4 fold more Zn was accumulated in the roots of the NHE. Approximately 2.7-fold more Zn was stored in the root vacuoles of the NHE, and thus became unavailable for loading into the xylem and subsequent translocation to shoot. Long-term efflux of absorbed 65Zn indicated that 65Zn activity was 6.8-fold higher in shoots but 3.7 fold lower in roots of the HE. At lower Zn levels (10 and 100 microM), there were no significant differences in 65Zn uptake by leaf sections and intact leaf protoplasts between the two ecotypes except that 1.5-fold more 65Zn was accumulated in leaf sections of the HE than in those of the NHE after exposure to 100 microM for 48 h. At 1,000 microM Zn, however, approximately 2.1-fold more Zn was taken up by the HE leaf sections and 1.5-fold more 65Zn taken up by the HE protoplasts as compared to the NHE at exposure times >16 h and >10 min, respectively. Treatments with carbonyl cyanide m-chlorophenylhydrazone (CCCP) or ruptured protoplasts strongly inhibited 65Zn uptake into leaf protoplasts for both ecotypes. Citric acid and Val concentrations in leaves and stems significantly increased for the HE, but decreased or had minimal changes for the NHE in response to raised Zn levels. These results indicate that altered Zn transport across tonoplast in the root and stimulated Zn uptake in the leaf cells are the major mechanisms involved in the strong Zn hyperaccumulation observed in S. alfredii H. PMID- 16362326 TI - Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides. AB - Theobroma cacao L. plants over-expressing a cacao class I chitinase gene (TcChi1) under the control of a modified CaMV-35S promoter were obtained by Agrobacterium mediated transformation of somatic embryo cotyledons. Southern blot analysis confirmed insertion of the transgene in eight independent lines. High levels of TcChi1 transgene expression in the transgenic lines were confirmed by northern blot analysis. Chitinase activity levels were measured using an in vitro fluorometric assay. The transgene was expressed at varying levels in the different transgenic lines with up to a sixfold increase of endochitinase activity compared to non-transgenic and transgenic control plants. The in vivo antifungal activity of the transgene against the foliar pathogen Colletotrichum gloeosporioides was evaluated using a cacao leaf disk bioassay. The assay demonstrated that the TcChi1 transgenic cacao leaves significantly inhibited the growth of the fungus and the development of leaf necrosis compared to controls when leaves were wound inoculated with 5,000 spores. These results demonstrate for the first time the utility of the cacao transformation system as a tool for gene functional analysis and the potential utility of the cacao chitinase gene for increasing fungal pathogen resistance in cacao. PMID- 16362327 TI - ZmLrk-1, a receptor-like kinase induced by fungal infection in germinating seeds. AB - We report here on the identification and characterization of ZmLrk-1, a member of the Lrk class of receptor-like kinases in Zea mays. This gene was found to be located at the bin21.40 region on the short arm of maize chromosome 8, closely linked to the previously reported pseudogene of the same class psiZmLrk (originally called Zm2Lrk). Transient expression experiments in onion epithelium cells, using a ZmLrk-1:GFP fusion protein, indicate that ZmLrk-1 is a membrane protein. ZmLrk-1 is ubiquitously expressed in the maize plant, including roots and aerial parts. In seeds, ZmLrk-1 transcripts can be detected by in situ hybridization exclusively at the basal endosperm transfer cell layer during the first stages of development. However, from 14 days after pollination its transcripts are preferentially detected at the upper half of the kernel, including both the aleurone and the starchy endosperm. ZmLrk-1 expression is not induced after treatment with salicylic acid, jasmonic acid or wounding, but it clearly increases after infection of germinating seeds with Fusarium oxysporum. This suggests that ZmLrk-1 could be involved in a sensing system to activate plant defence mechanisms against fungal attacks during endosperm development and seed germination. PMID- 16362328 TI - Expression profiling of the Arabidopsis ferric chelate reductase (FRO) gene family reveals differential regulation by iron and copper. AB - The Arabidopsis FRO2 gene encodes the iron deficiency-inducible ferric chelate reductase responsible for reduction of iron at the root surface; subsequent transport of iron across the plasma membrane is carried out by a ferrous iron transporter (IRT1). Genome annotation has identified seven additional FRO family members in the Arabidopsis genome. We used real-time RT-PCR to examine the expression of each FRO gene in different tissues and in response to iron and copper limitation. FRO2 and FRO5 are primarily expressed in roots while FRO8 is primarily expressed in shoots. FRO6 and FRO7 show high expression in all the green parts of the plant. FRO3 is expressed at high levels in roots and shoots, and expression of FRO3 is elevated in roots and shoots of iron-deficient plants. Interestingly, when plants are Cu-limited, the expression of FRO6 in shoot tissues is reduced. Expression of FRO3 is induced in roots and shoots by Cu limitation. While it is known that FRO2 is expressed at high levels in the outer layers of iron-deficient roots, histochemical staining of FRO3-GUS plants revealed that FRO3 is predominantly expressed in the vascular cylinder of roots. Together our results suggest that FRO family members function in metal ion homeostasis in a variety of locations in the plant. PMID- 16362329 TI - Expression profiling of starch metabolism-related plastidic translocator genes in rice. AB - The genes encoding the major putative rice plastidic translocators involved in the carbon flow related to starch metabolism were identified by exhaustive database searches. The genes identified were two for the triose phosphate/phosphate translocator (TPT), five for the glucose 6 phosphate/phosphate translocator (GPT) including putatively non-functional ones, four for the phosphoenolpyruvate/phosphate translocator (PPT), three for the putative ADP-glucose translocator (or Brittle-1 protein, BT1), two for the plastidic nucleotide transport protein (NTT), and one each for the plastidic glucose translocator (pGlcT) and the maltose translocator (MT). The expression patterns of the genes in various photosynthetic and non-photosynthetic organs were examined by quantitative real-time PCR. OsBT1-1 was specifically expressed in the seed and its transcript level tremendously increased at the onset of vigorous starch production in the endosperm, suggesting that the ADP-glucose synthesized in the cytosol is a major precursor for starch biosynthesis in the endosperm amyloplast. In contrast, all of the genes for OsTPT, OsPPT, and OsNTT were mainly expressed in source tissues, suggesting that their proteins play essential roles in the regulation of carbohydrate metabolism in chloroplasts. Substantial expression of the four OsGPT genes and the OspGlcT gene in both source and sink organs suggests that the transport of glucose phosphate and glucose is physiologically important in both photosynthetic and non photosynthetic tissues. The present study shows that comprehensive analysis of expression patterns of the plastidic translocator genes is a valuable tool for the elucidation of the functions of the translocators in the regulation of starch metabolism in rice. PMID- 16362330 TI - Secondary cell-wall assembly in flax phloem fibres: role of galactans. AB - Non-lignified fibre cells (named gelatinous fibres) are present in tension wood and the stems of fibre crops (such as flax and hemp). These cells develop a very thick S2 layer within the secondary cell wall, which is characterised by (1) cellulose microfibrils largely parallel to the longitudinal axis of the cell, and (2) a high proportion of galactose-containing polymers among the non-cellulosic polysaccharides. In this review, we focus on the role of these polymers in the assembly of gelatinous fibres of flax. At the different stages of fibre development, we analyse in detail data based on sugar composition, linkages of pectic polymers, and immunolocalisation of the beta-(1-->4)-galactans. These data indicate that high molecular-mass gelatinous galactans accumulate in specialised Golgi-derived vesicles during fibre cell-wall thickening. They consist of RG-I like polymers with side chains of beta-(1-->4)-linked galactose. Most of them are short, but there are also long chains containing up to 28 galactosyl residues. At fibre maturity, two types of cross-linked galactans are identified, a C-L structure that resembles the part of soluble galactan with long side chains and a C-S structure with short chains. Different possibilities for soluble galactan to give rise to C-L and C-S are analysed. In addition, we discuss the prospect for the soluble galactan in preventing the newly formed cellulose chains from completing immediate crystallisation. This leads to a hypothesis that firstly the secretion of soluble galactans plays a role in the axial orientation of cellulose microfibrils, and secondly the remodelling and cross-linking of pectic galactans are linked to the dehydration and the assembly of S2 layer. PMID- 16362331 TI - Repetition costs in word identification: evaluating a stimulus-response integration account. AB - Stimulus repetition facilitates performance in many experimental contexts. However, an episodic approach to interpreting repetition effects suggests that repetition effects should depend on how stimuli are encoded. In Experiments 1-3, stimulus repetition in a word identification task led to a cost rather than a benefit in performance, but only when the prime was presented for a relatively long duration. One account of these results is that long prime durations allow integration between stimulus and response codes to occur, which in turn can interfere with responding to a following identical target. In Experiment 4, a stimulus intensity based episodic specificity effect that was insensitive to the proportion of repeated trials supported this stimulus-response integration account. PMID- 16362332 TI - Hearing lips in a second language: visual articulatory information enables the perception of second language sounds. AB - We investigated the effects of visual speech information (articulatory gestures) on the perception of second language (L2) sounds. Previous studies have demonstrated that listeners often fail to hear the difference between certain non native phonemic contrasts, such as in the case of Spanish native speakers regarding the Catalan sounds /epsilon/ and /e/. Here, we tested whether adding visual information about the articulatory gestures (i.e., lip movements) could enhance this perceptual ability. We found that, for auditory-only presentations, Spanish-dominant bilinguals failed to show sensitivity to the /epsilon/-/e/ contrast, whereas Catalan-dominant bilinguals did. Yet, when the same speech events were presented audiovisually, Spanish-dominants (as well as Catalan dominants) were sensitive to the phonemic contrast. Finally, when the stimuli were presented only visually (in the absence of sound), none of the two groups presented clear signs of discrimination. Our results suggest that visual speech gestures enhance second language perception at the level of phonological processing especially by way of multisensory integration. PMID- 16362333 TI - Clinical significance of a wide excision policy for sacrococcygeal chordoma. AB - PURPOSE: Surgical treatment of sacrococcygeal chordoma is very difficult and the recurrence rate is high. We investigated the outcomes of wide excision in 12 sacrococcygeal chordoma patients treated between 1983 and 2003. METHODS: The 12 patients underwent primary wide excision. The conventional procedure was wide excision employing a chisel and airtome for sacral tumours, and in 1997 a new threadwire saw procedure was introduced. All 12 patients were included in a retrospective analysis of tumour- and treatment-related variables, and outcomes. RESULTS: Six patients underwent surgery with a combined anterior-posterior approach at the cephalad sacral border. In 12 patients, 11 lesions were broadly excised, and in the 1 remaining patient the intralesional margin showed tumour contamination due to a fracture macroscopically. The latter patient received post operative radiation therapy, but suffered a recurrence 8.0 years later. Three patients died before final follow-up: one from subsequent metastases, two elderly patients from complications. For the 10 chordoma patients, i.e. excluding the two who died due to complications, the 5, 10 and 20 years recurrence rates were 100, 66.7 and 66.7%, respectively. For the 12 patients, overall 5, 10 and 20 years survival rates were 83.3, 55.6 and 55.6%, respectively. CONCLUSIONS: Our results suggest that large chordoma should be widely excised, using a modified threadwire saw, with a combination of anterior-posterior procedures. PMID- 16362334 TI - 5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethylpyrazole acts in a reactive oxygen species-dependent manner to suppress human lung cancer growth. AB - PURPOSE: 5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethylpyrazole (SC 560) is a structural analog of celecoxib. Recent studies suggested that SC-560 inhibits the in vivo proliferation of colon and breast cancer cells to an extent similar to that observed in celecoxib, and that SC-560 exerts their growth inhibitory effects in a cyclooxygenase-independent manner. METHODS: In the current study, we sought to investigate the mechanism by which SC-560 inhibits the growth of human lung cancer cells. RESULTS: SC-560 more potently inhibited the growth of human A549, H460, and H358 lung cancer cell lines compared with that of human BEAS-2B normal bronchial epithelial cells. SC-560-induced growth inhibition was mainly due to the induction of cell-cycle arrest at the G1 phase without apoptosis induction. SC-560 rapidly and dose-dependently induced the generation of reactive oxygen species (ROS), followed by accumulation of cells at the G1 phase. Antioxidant pretreatment blocked the cell-cycle arrest and growth inhibition induced by SC-560. Combination treatment with other ROS-inducing agents such as alpha-tocopheryl succinate (TOS) augmented cellular response against SC-560, leading to synergistic apoptosis induction and growth suppression. Our data also showed that the apoptosis induced by combination treatment with SC-560 and TOS was mediated through ROS-dependent caspase activation. CONCLUSION: Collectively, our results demonstrate that SC-560 acts in a ROS-dependent manner to induce growth suppression in human lung cancer cells. PMID- 16362335 TI - Cathepsin B mediates TRAIL-induced apoptosis in oral cancer cells. AB - PURPOSE: The death ligand TRAIL (tumor necrosis factor-related apoptosis inducing ligand) triggers apoptosis in a variety of cancer cells, which implies the potential for therapeutic applications. The purpose of this study was to investigate the role of the lysosomal protease cathepsin B (CB) in mediating TRAIL-induced cell death in oral squamous cell carcinoma (OSCC) cells. METHODS: OSCC cell lines from primary tumor and lymph node metastasis were examined for expression of apoptosis markers by Western blots, enzyme activity assays, nuclear fragmentation assays, and FACS analysis. Gene-specific ribozymes or chemical inhibitors were used to inhibit CB or caspases in target cells. RESULTS: TRAIL induced activation of caspase-3, cleavage of Bid and poly-ADP-ribose polymerase, release of cytochrome c, and DNA fragmentation were blocked either by a pan caspase inhibitor (zVAD-fmk) or a CB inhibitor (CA074Me), consistent with the involvement of TRAIL as well as CB in cell death. The primary tumor cells were more susceptible to apoptosis than their corresponding lymph node metastatic cells. Stable transfection of a ribozyme which inhibited CB expression also decreased the apoptotic process. CONCLUSIONS: We conclude that TRAIL-induced apoptotic cell death in OSCC cells is mediated through CB or through caspase activation. Our data point to a new tumor-suppressive role for CB in OSCC which is opposed to the invasion- and metastasis-promoting functions of lysosomal proteases. PMID- 16362336 TI - Antibodies against Leishmania cross-react with Crithidia luciliae: indirect immunofluorescence and Dot-ELISA study in dogs. AB - Leismaniasis is zoonotic parasitic disease distributed in many areas of Mediterranean basin. Because of its pathogenicity and difficulty to grow in vitro, we used Leishmania infantum as primary and Crithidia luciliae as secondary source of antigen. We compared 100 canine sera by indirect immunofluorescence and Dot-ELISA. Both atigens showed same results. PMID- 16362337 TI - Invasion of the intestinal tract by sporozoites of Eimeria coecicola and Eimeria intestinalis in naive and immune rabbits. AB - Naive and immune specific-pathogen-free rabbits were inoculated in the duodenum with sporocysts of Eimeria coecicola or Eimeria intestinalis. Samples were taken from the following tissues: duodenum (site of penetration of sporozoites), ileum (specific target site of the endogenous development of E. intestinalis), vermiform appendix (target site of E. coecicola) and two extraintestinal sites, mesenteric lymph nodes (MLNs), and spleen. The presence of sporozoites was checked by immunohistochemistry. In rabbits primary-infected with E. coecicola, large numbers of sporozoites were detected in the duodenum, extraintestinal sites, and vermiform appendix. The abundance of sporozoites in the spleen, MLN, and appendix was significantly reduced in the immune rabbits, and the migration seemed impeded. In the rabbits infected with E. intestinalis, sporozoites were absent in the spleen and MLN, indicating that the route of migration is different from that of E. coecicola. The number of sporozoites in the crypts of the ileum was markedly reduced in the immune animals. PMID- 16362338 TI - First report and characterization of adult Uncinaria spp. in New Zealand Sea Lion (Phocarctos hookeri) pups from the Auckland Islands, New Zealand. AB - Two species of hookworms (Uncinaria lucasi and Uncinaria hamiltoni) have been formally described from pinnipeds, but dissimilar types are noted from these hosts. This report is the first description of hookworms (Uncinaria spp.) from the New Zealand sea lion, Phocarctos hookeri. The nematodes were collected from dead pups on Enderby Island (Auckland Islands, 50 degrees 30', 166 degrees 17') during January and February, 2004. Standard measurements of male and female hookworms were obtained, providing a general morphometric characterization of the hookworm species in P. hookeri. Considerable variations in the body length of adult hookworms were noted within the same host. The arrangement of some of the bursal rays differs from that described for U. lucasi and U. hamiltoni. PMID- 16362339 TI - First studies on the susceptibility of Omphiscola glabra (Gastropoda: Lymnaeidae) from central France to Fascioloides magna. AB - Experimental infections of Omphiscola glabra (preadult snails), originating from central France, to a Czech isolate of Fascioloides magna miracidia were carried out to determine if the local populations of O. glabra may ensure the larval development of this parasite and to compare these results with those noted for a natural snail host, Galba truncatula. The presence of experimentally infected snails was noted in the six populations of snails studied. However, only a few snails shed their cercariae (O. glabra 5.3 to 17.1%, G. truncatula 15.1% in the first population, and no shedding in the other). The shell heights of cercariae shedding (CS) snails were significantly greater than those of other infected snails, for O. glabra as well as for G. truncatula. The number of metacercariae noted in each snail group was low and showed insignificant variations. When experimental infections of O. glabra were performed in relation to the shell height of snails (from 1 to 14 mm) at miracidial exposure, the prevalence of infected snails significantly decreased with increasing shell heights at exposure. However, the presence of CS snails was only noted from the 5-6 to the 9 10 mm groups, and the mean number of metacercariae per group ranged from 27 to 44.2. Despite the high infectivity of the Czech isolate of F. magna miracidia, there was an incomplete adaptation with the French G. truncatula and O. glabra used in this study, as the metacercarial production was low, and cercarial shedding only occurred for snails which showed a strong increase of their shell height during F. magna infections. PMID- 16362340 TI - PROTOGIM: a novel tool to search motifs and domains in hypothetical proteins of protozoan genomes. AB - Whole sequencing of protozoan trypanosomatid genomes revealed the presence of several predicted unknown genes coding for hypothetical proteins. Pairwise, alignment-based, computational methods available online are unable to identify the function of these sequences. To detect clues to identify the function of hypothetical proteins, a user-friendly, bioinformatic tool named PROTOzoan Gene Identification Motifs (PROTOGIM, available on http://www.biowebdb.org/protogim ) was developed, which allows the user to search functional patterns of hypothetical proteins through the screening of regular expression in the sequences. The analysis of 1,194 trypanosomatid hypothetical proteins through PROTOGIM resulted in an identification of motifs and domains in 98% of the cases, demonstrating the reliability and accuracy of the employed method. The added value of this tool is the possibility to modify or insert new regular expressions to perform an analysis against either one or several sequences at the same time. An in silico strategy along with biochemical and molecular characterizations creates new possibilities to find the functions of hypothetical proteins at the postgenome era. PMID- 16362341 TI - Ultrastructural assessment of Plasmodium falciparum in age-fractionated thalassaemic erythrocytes. AB - Culture of Plasmodium falciparum in age-fractionated thalassaemic red blood cells (RBC) has shown evidence of parasite damage on light microscopy in older cells during the third culture cycle (96-144 h). In this report, parasites growing in thalassaemic trait and normal RBC were examined ultrastructurally from 96 to 144 h. All parasite stages in old thalassaemic RBC showed evidence of damage worsening with culture duration. There were cytoplasmic alterations with ribosomal damage, and parasite cytoplasm became increasingly loose and grainy, with multiple fissures. Discontinuity of the nuclear membrane with an abnormal nucleolus was seen at l20 h. Cytosomes remained normal, but damage to the food vacuole and shrunken disintegrating parasites were observed at 144 h. These changes are compatible with cellular degeneration and developmental retardation and would account for the schizont maturation arrest and reduced reinvasion rates previously reported. Increased free radicals associated with thalassaemic erythrocytes would explain these changes, further supporting the role for oxidant stress in the protective mechanism. PMID- 16362342 TI - Effect of Angiostrongylus costaricensis extract on eosinophilic pulmonary response in BALB/c mice. AB - Epidemiological and experimental studies have demonstrated an association between parasitic infections and the allergic diseases. A protective effect in asthma was shown in animals infected with helminths. The aim of this study was to determine the effect of Angiostrongylus costaricensis extract on inflammatory lung response to ovalbumin (OVA) in mice. Four BALB/c mice received A. costaricensis extract by intraperitoneal (i.p.) injection on the first day. Mice were immunised against OVA by i.p. injection on day (D) 5 and D12 and received a daily intranasal OVA challenge (40 microl) between the D19 and D21. On D23, we performed a bronchoalveolar lavage (BAL) on the mice. Four BALB/c mice (control group) were immunised against OVA using the same protocol, but did not receive parasite extract. Total cell counts (TCC) and differential cell counts were performed in BAL fluid samples. Eosinophil cell counts in BAL fluid were lower in the group that received A. costaricensis extract when compared with the control group (0.04 x 10(6) cells/ml and 0.01 x 10(6) cells/ml, respectively; p=0.04). TCC were not different between the groups studied. A. costaricensis extract in mice decreases eosinophilic response to OVA in BAL fluid. PMID- 16362343 TI - Interaction of beta-giardin with the Bop1 protein in Giardia lamblia. AB - A cDNA clone encoding a putative Bop1 homologous protein was identified in Giardia lamblia. Since Bop1 is a nucleolar protein involved in rRNA processing, thereby controlling the cell cycle, we investigated components of cell cycle control in G. lamblia by identifying the protein(s) that interact with Bop 1. Through an immunoaffinity column made with polyclonal antibodies specific to the recombinant Bop1 of G. lamblia, a pool of proteins was obtained from the crude extracts of Giardia and then used as antigens to immunize rats. By employing the resultant sera for cDNA library immunoscreening, we isolated cDNA clones encoding an immunopurified protein, which turned out to contain the gene for beta-giardin, a Giardia-specific cytoskeletal protein. The interaction between Bop1 and beta giardin was confirmed via two different methods, yeast two-hybrid assay and coimmunoprecipitation. PMID- 16362344 TI - First steps in antenatal diagnosis, 1956. AB - The original observation by the Canadians Barr and Bertram (Nature 163:676-677, 1949), that the nuclei of female cells from several species contained a small body, the sex chromatin, opened the possibility of sex determination in human epithelial and haematological cells. 7 years later it further led to the scientific question, whether or not human amniotic cells were sufficiently well preserved and numerous that they could form the basis for a reliable sex diagnosis of the human fetus. A technical condition was to test the safety and reliability of the physical access to the amniotic cavity, which needed co operation between geneticists and obstetricians. The article describes the first Danish steps in 1956 that led to the subsequent success of antenatal diagnosis as part of clinical genetics. PMID- 16362345 TI - Searching for signals of evolutionary selection in 168 genes related to immune function. AB - Pathogens have played a substantial role in human evolution, with past infections shaping genetic variation at loci influencing immune function. We selected 168 genes known to be involved in the immune response, genotyped common single nucleotide polymorphisms across each gene in three population samples (CEPH Europeans from Utah, Han Chinese from Guangxi, and Yoruba Nigerians from Southwest Nigeria) and searched for evidence of selection based on four tests for non-neutral evolution: minor allele frequency (MAF), derived allele frequency (DAF), Fst versus heterozygosity and extended haplotype homozygosity (EHH). Six of the 168 genes show some evidence for non-neutral evolution in this initial screen, with two showing similar signals in independent data from the International HapMap Project. These analyses identify two loci involved in immune function that are candidates for having been subject to evolutionary selection, and highlight a number of analytical challenges in searching for selection in genome-wide polymorphism data. PMID- 16362346 TI - Polymorphic micro-inversions contribute to the genomic variability of humans and chimpanzees. AB - A combination of inter- and intra-species genome comparisons is required to identify and classify the full spectrum of genetic changes, both subtle and gross, that have accompanied the evolutionary divergence of humans and other primates. In this study, gene order comparisons of 11,518 human and chimpanzee orthologous gene pairs were performed to detect regions of inverted gene order that are potentially indicative of small-scale rearrangements such as inversions. By these means, a total of 71 potential micro-rearrangements were detected, nine of which were considered to represent micro-inversions encompassing more than three genes. These putative inversions were then investigated by FISH and/or PCR analyses and the authenticity of five of the nine inversions, ranging in size from approximately 800 kb to approximately 4.4 Mb, was confirmed. These inversions mapped to 1p13.2-13.3, 7p22.1, 7p13-14.1, 18p11.21-11.22 and 19q13.12 and encompass 50, 14, 16, 7 and 16 known genes, respectively. Intriguingly, four of the confirmed inversions turned out to be polymorphic: three were polymorphic in the chimpanzee and one in humans. It is concluded that micro-inversions make a significant contribution to genomic variability in both humans and chimpanzees and inversion polymorphisms may be more frequent than previously realized. PMID- 16362347 TI - Detecting genome wide haplotype sharing using SNP or microsatellite haplotype data. AB - Genome wide association studies using high throughput technology are already being conducted despite the significant hurdles that need to be overcome (Nat Rev Genet 6:95-108, 2005; Nat Rev Genet 6:109-118, 2005). Methods for detecting haplotype association signals in genome wide haplotype datasets are as yet very limited. Much methodological research has already been devoted to linkage disequilibrium (LD) fine mapping where the focus is the identification of the disease locus rather than the detection of a disease signal. Applications of these approaches to genome wide scanning are limited by the strong model assumptions of the sharing process, which lead to computational complexity. We describe a new algorithm for the initial identification of disease susceptibility loci in genome wide haplotype association studies. Excess sharing of ancestral haplotypes, which indicates the presence of a disease locus, is detected with a simple, easy to interpret, chi2 based statistic. The method allows genome wide scanning for qualitative traits within reasonable computational timeframes and can serve as a first pass analysis prior to the usage of likelihood based methods, providing candidate regions and inferred susceptibility haplotypes. Our method makes no assumptions regarding the population history or the pattern of background LD. Statistical significance is evaluated with permutation tests. The method is illustrated on simulated and real data where it is applied to simple (cystic fibrosis) and complex disease (multiple sclerosis) examples. The statistic has low type I error and greater power to map disease loci over conventional single marker tests for low to moderate levels of LD. PMID- 16362349 TI - [The question of surgical therapy for necrolytic migratory erythema, a cutaneous disease]. AB - Necrolytic migratory erythema (NME) is a rare paraneoplastic dermatologic condition. Its underlying cause is usually a pancreatic islet cell tumour with marked glucagon secretion. The glucagonoma syndrome is characterised by pancreatic neuroendocrine neoplasm, NME, and diabetes mellitus. We present a case of glucagonoma syndrome in a 58-year-old woman with a history of recurrent cutaneous manifestations who was referred for surgical resection of a pancreatic neoplasm after the NME was finally diagnosed. We discuss diagnostic methods, differential diagnosis, and therapeutic management of this disease. PMID- 16362348 TI - Functional polymorphisms of FGA, encoding alpha fibrinogen, are associated with susceptibility to venous thromboembolism in a Taiwanese population. AB - To determine the genetic risk factors for venous thromboembolism (VTE), this study examined 14 genetic variants from 10 hemostatic genes in 186 Taiwanese VTE patients and the same number of matched controls, which demonstrated FGA (encoding alpha fibrinogen) Thr312Ala polymorphism was the only variant significantly associated with VTE. Nine genetic polymorphisms on the fibrinogen cluster region of chromosome 4q28 were further studied, in which four FGA polymorphisms were found in strong linkage disequilibrium and were significantly associated with VTE by genotype and allele frequency analyses. Haplotype analysis showed significantly different FGA haplotype frequencies between VTE patients and controls with the haplotype F1, containing -1051G, -3A, 312Ala and TaqI duplication alleles, significantly associated with susceptibility to VTE (P = 0.001). Haplotype-pair analysis results also indicated a strong association of the haplotype-pair F1F1 with VTE in various VTE patient subgroups. In vitro functional analysis indicated that FGA -1051G, -3A and TaqI duplication alleles enhanced significantly the transcription level of FGA; however, control subjects with FGA genotypes containing these alleles had no elevated plasma fibrinogen levels. In conclusion, our experimental data indicated that functional genetic variants in FGA are risk factors for VTE in Taiwanese populations. Determination of FGA genotypes will likely contribute to primary prevention of this condition. PMID- 16362350 TI - [Extracranial aneurysm of the carotid artery in a 12-year-old girl]. AB - Extracranial aneurysms of the carotid artery in childhood are rare, and we found just nine cases described in the literature within the last 20 years. We report on the clinical appearance, diagnosis, and therapy of a carotid artery aneurysm in a 12-year-old girl and review the literature. PMID- 16362351 TI - [Minimally invasive adrenalectomy for pheochromocytoma: routine or risk?]. AB - BACKGROUND: As a result of intraoperative catecholamine secretion with hemodynamic changes, larger tumor size, and marked neovascularization, adrenalectomy is more challenging and prone to complications for pheochromocytoma than for other adrenal diseases. The aim of this study was to examine the relative intraoperative risk of cardiovascular complications with the minimally invasive approach. PATIENTS AND METHODS: Between February 1992 and May 2005, 82 operations were performed on 71 pheochromocytoma patients at our clinic. Of them, nine adrenalectomies were bilateral, one was trilateral, and two patients had recurrent procedures. Another five patients were included whose first operations took place before 1992. Thirty-six procedures were carried out conventionally (35 transperitoneally, one retroperitoneally) and 46 were endoscopic (28 transperitoneally, 18 retroperitoneally). There was no conversion to open procedure. RESULTS: The median age at the time of surgery was 45 (24-75) years, and the median history of symptoms was 12 months (0-180). The openly resected pheochromocytomas were significantly larger than those in endoscopic operations: 5.5 cm (1-19 cm) vs 3.5 cm (0.5-8 cm) (P=0.0011). Compared with patients undergoing conventional procedures, those operated on endoscopically showed higher intraoperative systolic and diastolic blood pressures and peaks of more than 200 mmHg, although these differences were statistically insignificant. Multivariate analysis identified gender (P=0.0107), operative approach (P=0.0153), age (P=0.0364), and tumor size (P=0.0484) as factors with a possible influence on intraoperative hemodynamic alterations. Postoperative hospital stay was significantly shorter following endoscopic adrenalectomy (P=0.0001). CONCLUSION: Endoscopic adrenalectomy for pheochromocytoma is suitable as a routine operation and harbors no increased risk of cardiovascular complications, making it the method of choice. The open procedure should be reserved for extraadrenal tumors or large tumors with the suspicion of malignancy. PMID- 16362352 TI - [Trauma mechanism and diagnosis of blunt diaphragmatic rupture]. AB - BACKGROUND: Indirect diaphragmatic rupture (DR) is defined as acquired complete split of the diaphragm after blunt trauma and occurs in 1-5% of massively traumatized patients. The diagnosis is often difficult. However, particular trauma mechanisms and associated injuries may suggest the existence of DR. METHODS: A retrospective study was done of 21 consecutive patients from 1993 to 2004 at an university hospital. RESULTS: This study revealed a striking combination of DR with pelvic fractures (12/21, 57%). Of 19 polytraumatized patients, diagnosis was made initially in 13 (68%) and with a delay in six (32%). Right-sided injuries were common (8/21, 38%). Thus a 'protective' function of the liver does not exist. Diaphragmatic rupture can be missed in ventilated patients until extubation leads to herniation of abdominal organs into the thorax. CONCLUSIONS: Diaphragmatic rupture should be excluded in all severely abdominally traumatized patients, in particular those with concomitant pelvic fracture. Computed tomography with oral contrast medium is diagnostic in most cases. Access by laparotomy should be preferred after acute trauma, since additional intra abdominal injuries are frequent. PMID- 16362353 TI - [Current treatment for dry eye syndrome]. AB - The therapy of dry eye disease as a chronic problem necessitates a long-term therapeutic association between patient and ophthalmologist. The aim of therapy is to relieve symptoms and improve vision as far as possible with an acceptable level of effort on the part of the patient. Therapy of dry eye follows a staged concept including tear replacement, serum eye drops, punctum plugs, novel anti inflammatory drugs (cyclosporin A), and surgical procedures. Interdisciplinary care, especially of patients with autoimmune disease, is recommended and is best provided in a specialized dry eye outpatient clinic with quality control. PMID- 16362354 TI - Endurance training modulates the muscular transcriptome response to acute exercise. AB - We hypothesized that in untrained individuals (n=6) a single bout of ergometer endurance exercise provokes a concerted response of muscle transcripts towards a slow-oxidative muscle phenotype over a 24-h period. We further hypothesized this response during recovery to be attenuated after six weeks of endurance training. We monitored the expression profile of 220 selected transcripts in muscle biopsies before as well as 1, 8, and 24 h after a 30-min near-maximal bout of exercise. The generalized gene response of untrained vastus lateralis muscle peaked after 8 h of recovery (P=0.001). It involved multiple transcripts of oxidative metabolism and glycolysis. Angiogenic and cell regulatory transcripts were transiently reduced after 1 h independent of the training state. In the trained state, the induction of most transcripts 8 h after exercise was less pronounced despite a moderately higher relative exercise intensity, partially because of increased steady-state mRNA concentration, and the level of metabolic and extracellular RNAs was reduced during recovery from exercise. Our data suggest that the general response of the transcriptome for regulatory and metabolic processes is different in the trained state. Thus, the response is specifically modified with repeated bouts of endurance exercise during which muscle adjustments are established. PMID- 16362355 TI - Arthroscopic treatment of the hip in early and midstage degenerative joint disease. AB - The purpose of this study was to present the technique of arthroscopic capsule release in patients with early and midstage degenerative joint disease of the hip. In 22 patients we performed an arthroscopic capsular release of the hip joint capsule with simultaneous synovectomy and percutaneous drilling of areas of bone edema. The age of the patient ranged from 28 to 65 years (mean 52 years). There were 14 male and 8 female patients. All patients had suffered from a significant reduction of quality of life. In 15 of the 22 patients hip arthroplasty was already indicated in another institution. The preoperative Harris hip Score of 12 patients was below 69 points, 8 patients had a score between 70 and 79 points and 2 patients had a score between 80 and 89 points. At the time of follow-up (mean 26 months after surgery) 1 patient scored below 69 points, 3 patients scored between 70 and 79 points in the Harris hip score. Two patients scored between 80 and 89 points and 16 patients reached a score between 90 and 100 points. Subjective and objective 18 of 22 patients showed clinical relevant improvement. There was no complication in this series. Four patients underwent hip arthroplasty between 6 months and 2 years after the arthroscopic procedure. Minimal invasive arthroscopic treatments seem to decrease the subjective and objective complaints in early and midstage degenerative arthritis of the hip. PMID- 16362356 TI - Arthroscopic fibular sesamoidectomy in the management of the sesamoid osteomyelitis. AB - This is the first report in the English literature discussing the arthroscopic approach in the management of the fibular sesamoid osteomyelitis. First metatarsophalangeal joint fibular sesamoidectomy was successfully performed using big toe arthroscopy. The morbidity of the open surgical sesamoidectomy may be minimized by the arthroscopic approach. The authors describe the surgical principles and the potential advantages of arthroscopic sesamoidectomy compared with the open approach. PMID- 16362357 TI - The popliteal cyst. AB - A popliteal cyst, originally called Baker's cyst, is a synovial fluid-filled mass located in the popliteal fossa. The most common synovial popliteal cyst is considered to be a distension of the bursa located beneath the medial head of the gastrocnemius muscle. Usually, in an adult patient, an underlying intra-articular disorder is present. In children, the cyst can be isolated and the knee joint normal. The anatomy, etiopathogenesis, clinical presentation, differential diagnosis, imaging and treatment modalities of the popliteal cyst are presented. The authors try to answer some questions dealing with this condition. Is the cyst isolated, can it be treated as such, is its origin always well-defined and does surgical excision provide a permanent cure? PMID- 16362358 TI - Extracorporeal shock wave therapy in runners with a symptomatic heel spur. AB - The aim of this paper is to assess the benefit to treat plantar fasciitis with low-dose energy extracorporeal shock wave therapy (ESWT) and the efficacy of such treatment to abate the painful symptoms allowing a rapid return to the running activity. Our study included 54 running athletes treated for plantar fasciitis associated with a heel spur who received four sessions (once weekly) of low-dose ESWT, and followed prospectively on average 45 days, 6 and 24 months after their last session. The clinical results were excellent in 59% of cases, good in 12%, satisfactory in 21% and distinctly unsatisfactory in 8%. No patient was observed a significant modification of the heel spur at the follow-up X-ray. The ultrasound examination at 24 months showed a disappearance of the inflammation signs in 61% of cases. A strong correlation between ultrasound improvement and clinical results were found. Low-energy ESWT seems to be a good mean to treat plantar fasciitis in runners with a 71% of good or excellent results and a persistent improvement lasting 24 months. A randomized multicentric study seems to be necessary to define the type of energy that should be used in the future to treat plantar fasciitis, in particular in the athletic patients, to allow a faster return to sports activities. PMID- 16362359 TI - Proximal tibiofibular joint synovial cyst--one pathology with three different presentations. AB - Synovial cysts of the proximal tibiofibular joint are less common than synovial cysts of the knee joint but may present in a similar fashion and may be difficult to diagnose clinically. We report three cases of such synovial cysts: (1) the synovial cyst presented as an asymptomatic lump distal to lateral joint line of the knee; (2) the synovial cyst presented as a mass fluctuating in size with intermittent symptoms; (3) a man with a large mass in proximal anterior leg and drop foot. The patients were operated. The first and the third patients were treated successfully without recurrence, and complete recovery of the proneal nerve in third case ensued. The synovial cyst recurred in the second case; however, the patient refused a second operation. Age distribution and clinical manifestation of extraneural proximal tibiofibular joint synovial cyst is discussed in the light of relevant literature. PMID- 16362360 TI - Two-year follow-up on joint stability and muscular function comparing rotating versus fixed bearing TKR. AB - Mobile and fixed bearing in total knee replacement are still discussed controversially. In a retrospective, matched-pair study, we investigated 40 patients with computer-assisted (BrainLAB) primary total knee replacement (PFC Sigma, DePuy) performed two years before. Twenty patients each received a mobile bearing and a fixed bearing. We compared Womac Score, Knee Society Score, postoperative ROM, fluoroscopic measurement of knee stability in flexion and extension and isokinetic muscle strength using a Biodex-3 dynamometer. Both groups showed similar results concerning WOMAC Score (total rotating bearing: 23.05; fixed bearing: 22.57), KSS (rotating bearing: 174.89; fixed bearing: 176.1). Isokinetic muscle force demonstrated statistically significant superior results for knee flexion in the rotating bearing group. Medio-lateral stability revealed statistically significant superior results for the rotating bearing compared to fixed bearing in extension (P>0.05). In flexion only lateral stability was superior (P>0.05). Two-year clinical follow-up after computer assisted total knee replacement resulted in good clinical outcome with high patient satisfaction. Statistically significant better values for the rotating platform group were found for the medio-lateral stability in extension and the peak flexion torque. PMID- 16362361 TI - Kinematic and muscle activity characteristics of multidirectional shoulder joint instability during elevation. AB - Alterations of shoulder motion have been suggested to be associated with shoulder disorders. The objective of this study was to perform a 3D motion analysis (kinematic and electromyographical) of skeletal elements and muscles of shoulder joint in patients with multidirectional instability. Fifteen patients with multidirectional instability and 15 normal controls were investigated during continuous elevation in the scapular plane. The spatial coordinates of 16 anatomical points of the shoulder to determine kinematical parameters were quantified by an ultrasound-based motion analyzer. The activities of 12 muscles were measured by surface electromyography. Kinematic characteristics of motion were identified by scapulothoracic, glenohumeral, and humeral elevation angles; range of angles; scapulothoracic and glenohumeral rhythm; scapulothoracis, glenohumeral, and scapuloglenoid ratios; and the relative displacement between the rotation centers of the humerus and the scapula. The electromyographical characteristics of motion were modeled by the on-off pattern of muscle activity. Significant alterations in kinematical parameters were observed between patients and asymptomatic volunteers. The anterior, posterior, and inferior dislocations of shoulders with multidirectional instability could be properly modeled by the relative displacement between the rotation centers of the scapula and humerus. The shorter activity by m. pectoralis maior and all three parts of m. deltoideus and longer activity by m. supraspinatus, m. biceps brachii, and m. infraspinatus assure the centralization of the glenuhumeral head of a shoulder with multidirectional instability. PMID- 16362363 TI - Smooth pursuit eye movements in children. AB - Smooth pursuit eye movements consists of slow eye movements that approximate the velocity of the eyes to that of a small moving target, so that target image is kept at or near the fovea. Little information on smooth pursuit is available in children. We used an infrared eye tracker to record smooth pursuit in 38 typically developing children, aged 8-19 years. Participants followed a visual target moving sinusoidally at +/-10 degrees amplitude, horizontally and vertically at 0.25 or 0.5 Hz. The mean horizontal smooth pursuit gains, the ratio of eye to target velocities, were 0.84 at 0.25 Hz and 0.73 at 0.5 Hz. Mean vertical smooth pursuit gains were 0.68 at 0.25 Hz and 0.45 at 0.5 Hz. Smooth pursuit gains were significantly lower for vertical in comparison to horizontal tracking, and for 0.5 Hz in comparison to 0.25 Hz tracking (P<0.0001). Smooth pursuit gains increased with age (P<0.01, Pearson's correlation tests), with horizontal gains attaining reported adult values by mid adolescence. Vertical gains had large variability among participants. The median phase, the time interval between eye and target velocities, varied between 39 and 86 ms. Phase was not influenced by age. We conclude that smooth pursuit gains are lower in children than gains reported in adults. Vertical pursuit gain is significantly lower than horizontal pursuit gain. Gains improve with age and approach adult values in mid adolescence. Children have larger phases than reported adults values indicating that prediction in the smooth pursuit system is less mature in children. PMID- 16362364 TI - Increased density of GABAA receptors in the superior temporal gyrus in schizophrenia. AB - The superior temporal gyrus (STG) is strongly implicated in the pathophysiology of schizophrenia, particularly with regards to auditory hallucinations. In a previous study we reported a decrease in the density of M1 and M2/M4 muscarinic receptors in the STG in schizophrenia. In this study, we investigated the density of GABA(A) receptors in the left STG of schizophrenia patients compared to control subjects. We used quantitative autoradiography to investigate the binding of the agonist [(3)H] muscimol to GABA(A )receptors in the STG. A significantly higher density of [(3)H] muscimol binding was observed in the upper three quarters of the STG grey matter (corresponding to layers I-IV) than in the lower one-quarter (layers V-VI) in both groups. A significant increase (about 30%, P<0.05) in binding of [(3)H] muscimol was clearly observed in schizophrenia patients compared to control subjects. There were no significant correlations between [(3)H] muscimol binding density and age, post-mortem interval, brain pH or final recorded antipsychotic drug use. These results suggest an increase of GABA(A) receptor densities in the STG of schizophrenia patients. PMID- 16362365 TI - The modulation of intermanual interactions during the specification of the directions of bimanual movements. AB - In two experiments bimanual movements with various combinations of target directions were studied by means of the timed-response procedure. The findings revealed an adaptive modulation of intermanual interactions during direction specifications depending on particular target directions. For symmetric movements intermanual correlations of movement directions are positive, indicating a symmetric coupling. For parallel movements the positive intermanual correlations, observed at short preparation intervals, turn into negative correlations as the time available for motor preparation increases. Biases of mean directions, that can be observed for movements to targets with different eccentricities, reflect one or the other kind of coupling, symmetrical for symmetric target directions and parallel for parallel target directions. These biases are static, that is, they are present at long preparation times, and they are phasically enhanced at shorter preparation intervals. The task-adaptive modulation of intermanual interactions is superposed on a basic symmetry bias. PMID- 16362366 TI - The effect of circulating nitric oxide level on axial bone mineral density in postmenopausal Turkish women with rheumatoid arthritis: A preliminary report. AB - The aim is to investigate the differences in the circulating nitric oxide (NO) levels of rheumatoid arthritis (RA) patients, healthy controls and osteoporotic (OP) patients. We also examined whether the circulating NO levels may be correlated with bone mineral density (BMD) in RA patients. Forty-five patients with RA, 30 healthy women and 30 osteoporotic patients were recruited from the outpatient clinic. All the subjects were female and postmenopausal. Serum NO levels were measured (Nitrite/Nitrate, calorimetric method 1746081, Roche diagnostics, Mannheim, Germany) and BMD was measured at the spine and hip using dual energy X-Ray absorbtiometry (DEXA, Norland XR-46). Height and weight were measured and body mass index was calculated. Circulating NO levels were significantly higher in RA patients than other groups. Moreover, the RA group showed significantly higher BMD at lumbar spine and femoral neck regions compared to osteoporotic patients. However, the RA group showed significantly lower BMD at all sites than the controls. There was no correlation between circulating NO levels and BMD in all groups. We suggest that, unlike postmenopausal osteoporosis, inflammation induced osteoporosis is associated with RA is characterised by relatively preserved bone mass at the axial bone regions, and circulating NO levels as a parameter or determinant of inflammation are not correlated with axial BMD in RA patients. PMID- 16362367 TI - A hypothesis for the origin and pathogenesis of rheumatoid diseases. AB - It is well established that a correlation exists between rheumatoid arthritis (RA) and microbial damage. Material analyses have suggested that bacteria may be causative agents. This study was undertaken to further characterize the microbial agent responsible for pathogenesis of RA. In order to investigate whether substances in moist building materials can affect human cartilage, extracts from moist building materials were analysed for microbial components. Exposure of chondrocyte cultures to extracts in vitro showed that they were damaging the cultures. A direct correlation between strength of damage and concentration of MMP3 demonstrated that the effect was dose-dependent. High quantities of LPS were detected in the extracts. Experiments after deactivation with Polymyxin B showed that LPS are the causative agents. The present study leads to the hypothesis that LPS may bind to procollagen, as they bind to scavenger receptors. This procollagen endotoxin complex may block tropocollagen synthesis. PMID- 16362368 TI - Two different transposable elements inserted in flavonoid 3',5'-hydroxylase gene contribute to pink flower coloration in Gentiana scabra. AB - Pink-flowered gentian plants (Gentiana scabra) have been bred from spontaneous mutations of blue-flowered gentian plants, but the formation mechanism(s) is unknown so far. To investigate the process, two independent pink-flowered gentian plant lines were analyzed by a molecular biological approach. HPLC analysis showed that petals of the blue-flowered cultivar contained a small amount of cyanidin derivatives and major delphinidin derivatives, whereas pink petals had only a small amount of cyanidin derivatives. To find the causal factor(s) of this change, we focused on flavonoid 3',5'-hydroxylase (F3',5'H), which is a key enzyme for delphinidin biosynthesis in the flavonoid biosynthetic pathway. Molecular analyses confirmed that the loss of delphinidin synthesis could be attributed to the insertions of different transposable elements in the F3',5'H gene in each independent pink-flowered gentian plant. Sequence analysis showed that these transposable elements were classified into an hAT superfamily and terminal-repeat retrotransposon in miniature (TRIM), by which normal F3',5'H transcripts were interrupted. Southern blot analysis indicated that they belong to high copy number elements and are also found in a related gentian species (G. triflora). These results suggest that the transposable elements inserted in F3',5'H are the source of the mutations and may also play a substantial role in the genomic evolution of the genus Gentiana. PMID- 16362369 TI - The 'interactome' of the Knr4/Smi1, a protein implicated in coordinating cell wall synthesis with bud emergence in Saccharomyces cerevisiae. AB - The integrity of the Saccharomyces cerevisiae cell wall requires a functional Pkc1-Slt2 MAP kinase pathway that contributes to transient growth arrest, enabling coordination of cell division with cell wall remodelling. How this coordination takes place is still an open question. Recently, we brought evidence that Knr4 protein, whose absence leads to several cell wall defects, may play a role in this function. Here, we show that Knr4 is a monomeric protein that exhibits an aberrant mobility on a SDS-gel electrophoresis and a non-globular structure. Furthermore, Knr4 is an unstable protein that is degraded as cells enter the stationary phase of growth, while its corresponding gene is constitutively expressed. In exponentially growing cells on glucose, Knr4 appeared to be present in a protein complex that migrates with an apparent Mw superior to 250 kDa. Using the TAP-tag methodology, nine potential partners of Knr4 were identified, which could be distributed into three biological processes. A first group consisted of Slt2 and Pil1, two proteins dedicated to cell wall maintenance and biogenesis. The second group comprised four proteins (Bud6, Act1, Cin8 and Jnm1) implicated in the establishment of cell polarity and bud integrity during mitosis. The last group contained four proteins (Asc1, Ubc1, Hsc82 and Gvp36) that probably deal with the stability/degradation of proteins. Deletion analysis revealed that the domain of interaction covered 2/3 of the Knr4 sequence on the N-terminal side. Moreover, the replacement of the two in vivo phosphorylated Ser(200) and Ser(203) by alanines led to a mutated protein with reduced protein interactions and a weaker complementation ability towards knr4 null mutant phenotypes. These results together with previous data from genome scale two-hybrid and synthetic interaction screens support the notion that Knr4 is a regulatory protein that participates in the coordination of cell wall synthesis with bud emergence, and that this function may be modulated by phosphorylation of this protein. PMID- 16362370 TI - A cluster of 11 CBF transcription factors is located at the frost tolerance locus Fr-Am2 in Triticum monococcum. AB - Due to the adverse effects of cold temperatures on winter wheat, frost tolerance is an important trait for breeding programs in regions with severe winters. Frost tolerance locus Fr-A(m)2 was recently discovered in diploid wheat (Triticum monococcum L.). This locus was mapped as a QTL on chromosome 5A(m) in the same region as a QTL for the level of transcription of the cold-regulated gene COR14b at 15 degrees C. A CBF transcription factor was mapped in the center of these two overlapping QTLs. However, since the CBF gene family in wheat has numerous members, it was possible that multiple CBF genes were present at Fr-A(m)2. To investigate this possibility we initiated a systematic characterization of the CBF family in T. monococcum. Here we report the molecular characterization of thirteen TmCBF genes. Nine of them were numbered according to the closest barley HvCBF gene, and the other four that have no clear barley orthologues were assigned numbers TmCBF15 to TmCBF18. TmCBF5 and TmCBF18 were mapped on T. monococcum chromosomes 7A(m) and 6A(m), respectively, and are thus not candidates for the Fr-A(m)2 gene. The remaining eleven TmCBF genes are clustered at the Fr A(m)2 locus within five different Bacterial Artificial Chromosome (BAC) clones. These BACs were mapped using a high-density map and recombination events were found between most BACs. Lines carrying these recombination events will be useful to identify which of the CBF genes is responsible for the differences in frost tolerance between the T. monococcum parental lines at the Fr-A(m)2 locus. PMID- 16362372 TI - Wide-cross whole-genome radiation hybrid mapping of the cotton (Gossypium barbadense L.) genome. AB - Whole-genome radiation hybrid mapping has been applied extensively to human and certain animal species, but little to plants. We recently demonstrated an alternative mapping approach in cotton (Gossypium hirsutum L.), based on segmentation by 5-krad gamma-irradiation and derivation of wide-cross whole genome radiation hybrids (WWRHs). However, limitations observed at the 5-krad level suggested that higher doses might be advantageous. Here, we describe the development of an improved second-generation WWRH panel after higher dose irradiation and compare the resulting map to the 5-krad map. The genome of G. hirsutum (n = 26) was used to rescue the radiation-segmented genome of G. barbadense (n = 26) introduced via 8- and 12-krad gamma-irradiated pollen. Viable seedlings were not recovered after 12-krad irradiation, but 8-krad irradiation permitted plant recovery and construction of a 92-member WWRH mapping panel. Assessment of 31 SSR marker loci from four chromosomes revealed that the 8-krad panel has a marker retention frequency of ca. 76%, which is approximately equivalent to the rate of loss in a low-dose animal radiation hybrid panel. Retention frequencies of loci did not depart significantly from independence when compared between the A and D subgenomes, or according to positions along individual chromosomes. WWRH maps of chromosomes 10 and 17 were generated by the maximum likelihood RHMAP program and the general retention model. The resulting maps bolster evidence that WWRH mapping complements traditional linkage mapping and works in cotton, and that the 8-krad panel complements the 5-krad panel by offering higher rates of chromosome breakages, lower marker retention frequency, and more retention patterns. PMID- 16362371 TI - Kinase Cak1 functionally interacts with the PAF1 complex and phosphatase Ssu72 via kinases Ctk1 and Bur1. AB - Protein kinases orthologous with Cak1 of Saccharomyces cerevisiae (ScCak1) appear specific to ascomycetes. ScCak1 phosphorylates Cdc28, the cyclin-dependent kinase (CDK) governing the cell cycle, as well as Kin28, Bur1 and Ctk1, CDKs required for the transcription process performed by RNA polymerase II (RNA Pol II). Using genetic methods, we found that Cak1 genetically interacts with Paf1 and Ctr9, two components belonging to the PAF1 elongation complex needed for histone modifications, and with Ssu72, a protein phosphatase that dephosphorylates serine 5 phosphate in the RNA Pol II C-terminal domain. We present evidence suggesting that the interactions linking Cak1 with the PAF1 complex and with Ssu72 are not direct but mediated via Ctk1 and Bur1. We discuss the possibility that Ssu72 intervenes at the capping checkpoint step of the transcription cycle. PMID- 16362373 TI - A 60 kDa COX1 protein in mitochondria of carrot irrespective of the presence of C terminal extensions in the cox1 reading frames. AB - An inverted repeat in the mitochondrial DNA of carrot mitochondrial type (mitotype) SW3 has been sequenced in the context of its up- and downstream genomic environments. The 3,159 bp repeat encodes rps7 and cox1, but the cox1 reading frame does not encode a stop codon at a position predicted by alignment with other plant cox1 genes; cox1 extends into both of the downstream flanking sequences as unique C-terminal extensions of 224 (cox1-1) or 284 (cox1-2) amino acids. Northern and RT-PCR analyses revealed that rps7 and cox1 are co transcribed and RNA-edited. In three additional mitotypes of carrot, rps7 and cox1 are co-transcribed, but are present as single copies within the genome. In mitotype FG17, the cox1 gene has a C-terminal extension like cox1-1, but in mitotypes FW2 and FW4, the cox1 genes lack C-terminal extensions and have stop codons and 3' UTRs like those of other plant cox1 genes. The C-terminal extensions are transcribed in SW3 and FG17 and result in longer primary transcripts than those from FW2 and FW4. Western immunoblotting of mitochondrial proteins from all mitotypes detected COX1 products of the predicted unextended size in all mitotypes, irrespective of the presence of a C-terminal extension in the reading frame. PMID- 16362376 TI - [Diagnostics and treatment of posterior cruciate ligament injuries]. AB - Over the last few years, the posterior cruciate ligament (PCL) has taken over from the anterior cruciate ligament (ACL) as the new challenge in knee diagnosis and treatment. PCL injuries are much more frequent than previously thought. Despite increasing experience, we are often still confronted with an inappropriate and delayed diagnosis of this injury and its concomitant lesions. Additionally, the outcome of surgical treatment has not yet reached an acceptable rate of satisfaction. Therefore, the goal of this concept review is to give a comprehensive insight into injury mechanisms, diagnostics and treatment regimes based on our experience and data from the international literature. Furthermore, diagnostic problems with respect to clinical examination and imaging techniques are discussed. We also present a differential concept for the perioperative and conservative management of PCL deficient knees in order to adequately address concomitant injuries such as posterolateral rotatory instability and combined ACL injuries, with the aim of further improving results. PMID- 16362375 TI - [Cardiovascular MRT--replacement of diagnostic invasive coronary angiography?]. AB - With cardiovascular magnetic resonance imaging (CMR), the necessity of invasive coronary angiography may be increasingly avoided. CMR provides information about the anatomy of the coronaries themselves (e.g. anomalies, aneurysm), functional information on myocardial blood flow (dobutamine-stress-MR, perfusion measurement) and detailed information on cell-mediated alterations (e.g. fibrosis, necrosis). However, visualization of distal coronary vessels and the small side branches is not yet adequate, so that complete replacement of invasive coronary angiography by CMR is not possible. PMID- 16362378 TI - [Fetal magnetic resonance imaging. Diagnostics in congenital diaphragmatic hernia]. AB - Ultrasonography is the primary imaging modality for the evaluation of fetal or maternal anomalies. This method is safe, relatively inexpensive, easily accessible, and allows real-time imaging. Continuous technical improvements in ultrasonography in the last 10-15 years have led to improved diagnostic accuracy for fetal malformations. In cases of complex anomalies magnetic resonance imaging (MRI) can provide additional information. MRI has evolved as a valuable diagnostic method for evaluating fetal pathology. Particularly with regard to similarity of liver and lung parenchyma in ultrasonography, diagnosis of congenital diaphragmatic hernia (CDH) can be difficult. Beside morphological aspects, e.g., herniation of abdominal contents into the chest, small amounts of compressed lung can be visualized on MRI. The feasibility of using volumetric measurement on MRI may be helpful to predict high-risk fetuses and facilitate decisions to assure adequate prenatal and postnatal management to improve postnatal outcome. PMID- 16362380 TI - [MRI of the fetal abdomen]. AB - Magnetic resonance imaging (MRI) is an important diagnostic component for central nervous system and thoracic diseases during fetal development. Although ultrasound remains the method of choice for observing the fetus during pregnancy, fetal MRI is being increasingly used as an additional technique for the accurate diagnosis of abdominal diseases. Recent publications confirm the value of MRI in the diagnosis of fetal gastrointestinal tract and urogenital system diseases. The following report provides an overview of MRI-examination techniques for the most frequent diseases of the abdomen. PMID- 16362379 TI - [Indications for fetal magnetic resonance imaging]. AB - Indications to perform fetal magnetic resonance imaging (MRI) are composed of common ones related to methodological problems of ultrasound (US) assessment (such as for instance hydramnios) and special ones. The latter are related to MR capability of high-resolution soft tissue contrast and an extended field of view that allows visualization of the whole fetus, even in later stages of pregnancy. The most important indications include confirmation of US findings, work-up of malformations with respect to individual prognosis and genetic background, differentiation between acquired conditions and malformations, visualization of pathologies that have to be treated surgically immediately after birth, and morphological changes of the placenta. PMID- 16362381 TI - [On the uncertainty of measurements illustrated by the BONN Risk Index]. AB - Every result of a measurement is subject to error. This error is inevitably caused by inaccuracies of the devices used and by environmental influences. For correct evaluation of a measurement result, this error must be estimated as exactly as possible. In kidney stone research and urolithiasis treatment, risk indices are applied to determine a patient's health status [e.g., AP(CaOx), RS(CaOx), BRI]. These operands (indices) combine important physical and chemical parameters of a certain disease pattern in a theoretically or empirically obtained mathematical formula. The index-related total error resulting from the single errors of the individual input parameters might be calculated with the Gaussian error propagation method. This method takes into account that individually determined parameters contribute differently to the total result. Due to this, summation of the various specific errors is insufficient. Instead, they must contribute to the total error depending on their individual weight. In medical borderline cases, knowledge of the value of this error may improve diagnosis since critical observation of an acquired value helps to avoid false interpretation. Using the BONN-Risk-Index (BRI) as an example for determination of the urinary calcium oxalate crystallization risk in native urine, the method of error calculation is demonstrated in detail and the advantages of including discussion of error in method development are discussed. PMID- 16362382 TI - The management of infections due to drug-resistant gram-positive bacteria. PMID- 16362383 TI - Antibiotics and gastrointestinal colonization by vancomycin-resistant enterococci. AB - Although several classes of antimicrobial agents have been associated with colonization or infection with glycopeptide-resistant enterococci (GRE) in individual clinical studies, the agents most commonly implicated are extended spectrum cephalosporins and compounds with potent activity against anaerobic bacteria, including ticarcillin-clavulanic acid. In some clinical studies, formulary alterations designed to minimize the use of extended-spectrum cephalosporins or ticarcillin-clavulanic acid have resulted in significant decreases in colonization and infection by GRE. Experimental data using a mouse model of GRE gastrointestinal colonization indicate that persistence of high level GRE colonization of the mouse gastrointestinal tract is promoted by exposure to agents with potent activity against anaerobic bacteria, suggesting that reduction of competing flora is the major factor leading to persistence of high-level colonization. One study performed in humans is consistent with this model and suggests that high levels of colonization may promote spread of resistant organisms in the nosocomial setting. Establishing colonization with GRE in uncolonized mice correlates with exposure to agents that are (a) secreted into the bile in significant concentrations and (b) have negligible activity against the colonizing enterococcal strain. Differences between piperacillin-tazobactam and ceftriaxone in the establishment model can be attributed directly to differences in their anti-enterococcal activity. Modification of antimicrobial prescribing practices may play an important role in facilitating successful infection control efforts to limit GRE in the nosocomial setting. PMID- 16362384 TI - Total hip arthroplasty for developmental hip dysplasia. AB - We reviewed 38 hip replacements in 33 female patients (mean age 55.3 years) with developmental hip dysplasia. One patient had died and the remaining 32 patients (36 hips) had a mean follow-up of 12.2 years (range 8-19 years). All hips were replaced using the Muller cemented implant, and in 32 hips bulk femoral head autograft was used. In 33 hips the socket was reconstructed at the level of the true acetabulum. Complications included one intra-operative femoral fracture and two early dislocations. Correction of leg length discrepancy was possible in 30 patients. The post-operative mean modified Merle d'Aubigne and Postel scores for pain, movement and walking were 5.9, 5, and 5.3 respectively. One cup was revised due to aseptic loosening at ten years. All grafts united, but minor graft resorption was noticed in 24 hips, moderate in 2 hips and major in 1 hip. PMID- 16362385 TI - Distinct association of gene polymorphisms of estrogen receptor and vitamin D receptor with lumbar spondylosis in post-menopausal women. AB - Contribution of genetic backgrounds to the etiology of lumbar spondylosis has been suggested by epidemiological studies. This study was designed to determine the association of restriction fragment length polymorphisms (RFLPs) of estrogen receptor (ER), vitamin D receptor (VDR), parathyroid hormone (PTH) and interleukin-1beta (IL-1beta) genes with the radiological severity of lumbar spondylosis at the disk level from L1/2 to L5/S1 in Japanese post-menopausal women. ER and VDR RFLP haplotypes were associated with the severity of spondylosis in the upper levels (L1/2 and L2/3) more than in the lower levels. Association of ER genotype was more pronounced in the group younger than average than in the older group, while that of VDR genotype was more significant in the older group. Neither PTH nor IL1-beta RFLP was associated with the severity at any levels in either stratified group. We thus conclude that ER and VDR genes may contribute to lumbar spondylosis in a distinct manner: estrogen sensitivity influences the severity in the early phase after menopause while vitamin D plays an important role at older ages when the contribution of estrogen loss is weaker. PMID- 16362386 TI - Renal cement embolism during percutaneous vertebroplasty. AB - Percutaneous vertebroplasty (PVP) is an effective treatment for lesions of the vertebral body that involves a percutaneous injection of polymethylmethacrylate (PMMA). Although PVP is considered to be minimally invasive, complications can occur during the procedure. We encountered a renal embolism of PMMA in a 57-year old man that occurred during PVP. This rare case of PMMA leakage occurred outside of the anterior cortical fracture site of the L1 vertebral body, and multiple tubular bone cements migrated to the course of the renal vessels via the valveless collateral venous network surrounding the L1 body. Although the authors could not explain the exact cause of the renal cement embolism, we believe that physicians should be aware of the fracture pattern, anatomy of the vertebral venous system, and careful fluoroscopic monitoring to minimize the risks during the PVP. PMID- 16362388 TI - [Sclerotherapy for varicosities]. AB - Sclerotherapy is a standard treatment for intradermal varicose veins and branch varicosities. In the treatment of insufficient saphenous veins, crossectomy and stripping used to be the methods of choice. In the last few years good results have also been reported for endovenous methods such as endovenous laser and radio frequency treatment. Sclerotherapy is more effective and even appropriate for larger saphenous veins when the sclerosing liquid is replaced by a foam agent. This cost-effective approach can be done on an outpatient basis without additional anesthesia. Prospective randomized studies are need to confirm the effectiveness. PMID- 16362389 TI - [Endolumninal catheter-guided laser therapy of varicosities]. AB - Chronic venous insufficiency is one of the most common diseases with a high socio economic importance. The most common operation is vein stripping to remove the greater or lesser saphenous vein eliminating venous reflux. In comparison to classical vein stripping, endoluminal laser therapy is a minimally invasive and effective alternative with few complications and a shorter convalescent period. Varicosities can also be treated with the laser, using local anesthesia and sonographic monitoring. The recurrence rate is around 2%, comparable to surgical methods. This article reviews the practical aspects of endoluminal laser therapy, concentrating on the two most widely available systems, reviews the current literature and considers other endoluminal approaches. PMID- 16362390 TI - Glomerulonephritis associated with chronic infection from long-term central venous catheterization. AB - There have been few reports on immune complex-mediated glomerulonephritis associated with chronic infection from long-term central venous catheterization in adulthood. We report here on a 13-year-old boy with nephritis who exhibited glomerulonephritis that had been induced by the long-term use of central venous catheters, and its resolution after extraction of the central venous catheter. A diagnosis of glomerulonephritis associated with chronic infection caused by long term central venous catheterization was made, based on the absence of clinical findings after removal of the catheter, hypocomplementemia, pathology findings resembling membranoproliferative glomerulonephritis, and detection of Staphylococcus epidermidis from culture of the removed catheter culture. For clinicians using long-term central venous access for parenteral feeding, rapid catheter exchange is necessary for patients with fever of unknown origin. PMID- 16362391 TI - Thin basement membrane nephropathy associated with minimal change disease in a 15 year-old boy. AB - Thin basement membrane nephropathy (TBMN) is characterized clinically by persistent hematuria, minimal proteinuria, normal renal function, another family member with hematuria, and a benign course. Especially in childhood TBMN, proteinuria of any degree is reported to be uncommon. We report on a boy with benign familial hematuria found by urinary screening at 3 years of age who presented with nephrotic syndrome (NS) at 15 years of age. His renal histology showed TBMN associated with minimal change disease (MCD). Treatment with corticosteroid resulted in complete remission of NS in a short period of time, while isolated hematuria persisted during the follow-up period despite this therapy. We speculate, therefore, that the nephrotic range proteinuria is not due to TBMN but rather is the manifestation of associated MCD. Several cases of TBMN with NS have been reported in adults, but it has not yet been reported in children in the literature. To our knowledge, this is the first case of childhood TBMN associated with NS resulting from coincidental MCD. PMID- 16362392 TI - Successful steroid-sparing treatment of renal limited sarcoidosis with mycophenolate mofetil. AB - Sarcoidosis is a multi-system disorder characterized by non-caseating epithelioid granulomas in multiple organs. The disease usually presents in young adults and is uncommon in children. Renal involvement can usually occur due to granulomatous interstitial nephritis, but renal failure is uncommon. Corticosteroids are the mainstay of therapy. We present the report of a child with severe renal failure secondary to renal limited sarcoidosis who was successfully treated with corticosteroid induction therapy. Because of the severe side effects of corticosteroids, mycophenolate mofetil was added and corticosteroids were tapered off. The child has been in sustained remission for over a year with mycophenolate mofetil monotherapy. PMID- 16362393 TI - An analysis of renal tubular acidosis by the Stewart method. AB - Renal tubular acidosis (RTA) comprises a group of disorders characterized by a low capacity for net acid excretion and persistent hyperchloremic, metabolic acidosis. To investigate the role of chloride, we performed hypotonic (0.45%) saline-loading experiments in 12 children with alkali-treated distal RTA (dRTA) and compared the results with data obtained from 17 healthy control subjects. In patients, but not in controls, saline loading induced both hyperchloremia and metabolic acidosis. Hyperchloremia was associated with high total and high distal fractional reabsorption of chloride [C(H20)/(C(H20)+C(Cl))]. The increase in plasma chloride varied inversely with the fractional excretion of chloride (C(Cl)) and correlated with the decrease in blood pH. However, the urinary excretion of bicarbonate did not correlate with either changes in blood pH or plasma bicarbonate concentration. Our findings suggest that the mechanism of hyperchloremia was enhanced Cl(-)/HCO(3) (-) exchange by the distal tubule. The resulting metabolic acidosis is better explained by changes in the strong ion difference (the Stewart theory) than by changes in the urine bicarbonate excretion (the traditional theory). PMID- 16362394 TI - [Rare entity of a macroscopically visible tumor cone of squamous cell carcinoma in the greater veins of the head and neck]. AB - Cones of malignant tumors are commonly known but only a few descriptions of cones in the head and neck area exist - as seen in a 91-year-old patient under our care. Cones of thymic and thyroid cancers are described in the literature. There are no descriptions of cones of head and neck squamous cell carcinomas. Metastases of squamous cell carcinoma in the head and neck area are very aggressive and have a high potential for vascular neogenesis. A vascular cone might be possible by vascular formation or by mechanical intrusion as described elsewhere. PMID- 16362395 TI - [CT assisted mandibular osteodistraction with reconstruction of the temporomandibular joint and two piece maxilla]. AB - BACKGROUND: The possibilities for computer assisted planning and its surgical conversion with stereolithographic templates are shown for complex maxillofacial surgical interventions. CASE REPORT: We describe the case of a patient with extreme micrognathia due to an ankylosis of the temporomandibular joint. In addition to the exact determination of the osteotomy lines on the mandibular angle and the segment osteotomy of the maxilla, vectors of the bidirectional osteodistraction were also predictable. CONCLUSION: In comparison with the conventional procedures, computer assisted planning achieved a much higher degree of precision and safety with reduced complication rates. The CMF module is a component of the SimPlant software which enables the precise planning of osteotomies and osteodistractions. PMID- 16362396 TI - [Acute mesenteric ischemia in a patient with relapsing polychondritis]. AB - We report about a 43-year old woman with relapsing polychondritis, admitted with progressive malaise, fatigue, anorexia and profound weight loss. Two years ago a nasal chondritis with characteristic changes of the nasal profil, scleritis, laryngitis and unspecific general symptoms (fever, fatigue, signs of a systemic inflammation) developed and relapsing polychondritis was diagnosed. The patient was treated initially with azathioprine followed by methotrexate in combination with ciclosporine and glucocorticoids. During the hospitalization her condition slowly worsened, and crampy abdominal pain developed subsequently with massive tenderness, rebound and guarding. The clinically presumed diagnosis of ileus and peritonitis was radiomorphologically confirmed with signs of enteric perforation and a laparatomy was performed. A complete occlusion of the A. mesenterica superior and stenosis of the truncus coeliacus were diagnosed followed by a revascularization with an aorto-mesenteric bypass and subsequent resection of the necrotic ischemic ileum. An association with various autoimmune disorders including vasculitic syndromes is well known for relapsing polychondritis. The case demonstrated a progressive mesenteric ischemia with the acute exacerbation caused by a vasculitic mesenterial occlusion. The unusual presentation should be considered in the differential diagnosis of uncommon abdominal symptoms during the course of relapsing polychondritis. PMID- 16362397 TI - Single-dose metronidazole vs 5-day multi-drug antibiotic regimen in excision of pilonidal sinuses with primary closure: a prospective, randomized, double-blinded pilot study. AB - AIM: To compare infection-related wound complications following excision of pilonidal sinuses with primary closure using either single-dose intravenous (i.v.) administration of metronidazole preoperatively or a broad-spectrum multi drug regimen. PATIENTS AND METHODS: This is a double-blinded study wherein 50 patients were randomized into receiving either single-drug (metronidazole 500 mg i.v.) prophylaxis preoperatively or multi-drug cover (cefuroxime 1.5 g i.v. and metronidazole 0.5 g i.v. preoperatively, and co-amoxiclav 375 mg orally 8-hourly postoperatively for 5 days). They were reviewed at 1, 2 and 4 weeks postoperatively. Wounds were graded as follows: I, healthy; II, redness and swelling of edges; III, abscess in relation to a suture; IV, spreading wound infection; and V, wound breakdown. Other factors considered were the distance from the lowest wound margin to the anal verge and previous local surgery. RESULTS: Fifty patients (38 men and 12 women, mean age 27 years) underwent pilonidal sinus surgery. At week 1, there was no difference in wound infection rates (p=0.9). However, there were significantly more wound infections in the single-drug group at week 2 (p<0.0001) and week 4 (p=0.03). Seventy-two per cent of all patients had complete wound healing at week 4. Distance from the anal verge and previous surgery did not affect wound infection rates (p> or =0.2). Treating such complications costs 73,219.20 dollars per 100 patients. CONCLUSION: A broad-spectrum 5-day regimen is superior to 'single-shot' antibiotic prophylaxis in preventing infection-related wound complications. However, this study needs to be conducted in a larger number of patients to have statistical power. PMID- 16362398 TI - Neutrophil apoptosis in autoimmunity. AB - The regulation of death pathways in neutrophils has been of long interest, but the relevance of neutrophil apoptosis to the pathogenesis and treatment of autoimmune diseases has only recently been becoming recognized. This brief review addresses the relevant pathways by which neutrophil apoptosis is regulated and summarizes the current understanding regarding the potential role of apoptotic neutrophils in the initiation and/or propagation of autoimmunity, as well as the applicability of modulation of neutrophil death in the treatment of autoimmune diseases. PMID- 16362399 TI - Initial deficit and recovery of function after MDMA preexposure in rats. AB - RATIONALE: 3,4-methylenedioxymethamphetamine (MDMA) exposure was reported to result in deficits in serotonergic neurotransmission with concomitant behavioral suppression and tolerance to MDMA. Some data have also suggested that the neurochemical deficits recover over time, raising the question as to whether behavioral suppression would show a similar recovery. OBJECTIVES: The possibility of recovery of behavioral deficits was examined in the present study. Rats were administered an MDMA pretreatment regimen that was shown to produce numerous serotonergic deficits and behavioral suppression 2 weeks thereafter. The full expression of MDMA-produced hyperactivity was dependent upon serotonergic integrity, therefore, the present study aimed to determine whether MDMA pretreated rats were tolerant to MDMA 2 weeks after exposure. Further, because serotonergic deficits have shown recovery over time, similar behavioral tests were conducted at a later time point to determine whether functional recovery was evident. METHODS: MDMA-produced hyperactivity was measured at different withdrawal periods (2 and 12 weeks) to determine initial effects and the possibility of recovery of function. RESULTS: In saline-pretreated control rats, +/-MDMA (0.0-10.0 mg/kg) produced a dose-dependent increase in locomotor activity. Rats that had received prior exposure to MDMA (4 x 10 mg/kg MDMA injections administered at 2 h intervals) demonstrated tolerance when the activity was measured 2 weeks after pretreatment. For these rats, there was a downward shift in the dose-effect curve for MDMA-produced hyperactivity. MDMA produced hyperactivity in rats that were tested 12 weeks after pretreatment was, however, comparable to controls, suggesting recovery of function. CONCLUSION: These data are consistent with the idea that high dose MDMA exposure produces neuroadaptations that exhibit recovery with extended abstinence from the drug. PMID- 16362400 TI - Effects of a brisk walk on blood pressure responses to the Stroop, a speech task and a smoking cue among temporarily abstinent smokers. AB - A review and meta-analysis by Hamer et al. (2006) showed that a single session of exercise can attenuate post-exercise blood pressure (BP) responses to stress, but no studies examined the effects among smokers or with brisk walking. Healthy volunteers (n=60), averaging 28 years of age and smoking 15 cigarettes daily, abstained from smoking for 2 h before being randomly assigned to a 15-min brisk semi-self-paced walk or passive control condition. Subject characteristics, typical smoking cue-elicited cravings and BP were assessed at baseline. After each condition, BP was assessed before and after three psycho-social stressors were carried out: (1) computerised Stroop word-colour interference task, (2) speech task and (3) only handling a lit cigarette. A two-way mixed ANCOVA (controlling for baseline) revealed a significant overall interaction effect for time by condition for both systolic blood pressure (SBP) and diastolic blood pressure (DBP). Univariate ANCOVAs (to compare between-groups post-stressor BP, controlling for pre-stressor BP) revealed that exercise attenuated systolic BP and diastolic BP responses to the Stroop and speech tasks and SBP to the lit cigarette equivalent to an attenuated SBP and DBP of up to 3.8 mmHg. Post exercise attenuation effects were moderated by resting blood pressure and self reported smoking cue-elicited craving. Effects were strongest among those with higher blood pressure and smokers who reported typically stronger cravings when faced with smoking cues. Blood pressure responses to the lit cigarette were not associated with responses to the Stroop and speech task. A self-paced 15-min walk can reduce smokers' SBP and DBP responses to stress, of a magnitude similar on average to non-smokers. PMID- 16362401 TI - Anxiolytic-like effects induced by nitric oxide synthase inhibitors microinjected into the medial amygdala of rats. AB - RATIONALE: The amygdaloid complex plays a central role in the neuroanatomical circuits that coordinate defensive responses. Nitric oxide (NO) has been involved in the neurochemical, hormonal, and behavioral changes related to stress and anxiety. A high density of NO-producing neurons is observed in the medial amygdala (MeA). These neurons are activated after exposure to threatening stimuli such as a live predator. OBJECTIVE: To test the hypothesis that microinjection into the MeA of two NO synthase inhibitors, NG-nitro-L-arginine methyl ester (L NAME) or 7-nitroindazole (7-NI), would produce anxiolytic effects. METHODS: Male Wistar rats (n=8-10/group) were submitted to stereotaxic surgery to implant bilateral stainless steel guide cannulae aimed at the MeA. Six days after the surgery, the animals received intra-MeA microinjections of the drugs or vehicle and, 10 min later, were submitted to the elevated plus-maze (EPM) or the light dark transition procedures. RESULTS: Both L-NAME (50-200 nmol) and 7-NI (5 and 10 nmol) increased open-arm exploration in the EPM without changing the number of enclosed arm entries, indicating an anxiolytic-like effect. The anxiolytic-like effect of L-NAME (200 nmol) was prevented by pretreatment with L-arginine (100 nmol). Injections of 7-NI or L-NAME outside the MeA did not produce any significant change in EPM exploration. When tested in the light-dark test, L-NAME (200 nmol) or 7-NI (10 nmol) increased the time when the animal remained in the light compartment of the light-dark box. CONCLUSIONS: The present results suggest that inhibition of NO formation in the MeA produces anxiolytic-like effect in rats. PMID- 16362402 TI - Nicotine and nonnicotine factors in cigarette addiction. AB - RATIONALE: A great deal of research supports the role of nicotine in cigarette addiction. However, the effectiveness of nicotine replacement therapy (NRT) as a smoking cessation treatment has fallen short of initial hopes. A key reason may be that NRT does not address nonnicotine components of smoking reinforcement. These include constituents that provide reinforcing sensory stimulation, components that minimize excessive irritation from inhaled nicotine and other pharmacologically active compounds in cigarette smoke. OBJECTIVE: Studies using various paradigms to dissociate nicotine from other components of smoking are summarized. RESULTS: Nonnicotine components provide many rewarding effects, often surpassing the direct effects of nicotine. Substitutes for the sensory effects of smoking may be effective in relieving craving for cigarettes and in facilitating smoking cessation. Moreover, techniques for devaluing smoking-related cues may decrease craving and enhance subsequent abstinence. Promising approaches for devaluing smoke cues include extinction-based treatments employing denicotinized cigarettes and the use of nicotinic agonist and/or antagonist treatment during the weeks leading up to a quit attempt. Recent studies suggest that incorporating these approaches into a treatment program may significantly increase smoking abstinence rates. Preliminary findings also suggest that replacement of the effects of monoamine oxidase inhibitors contained in cigarette smoke may enhance quit rates. CONCLUSIONS: While current NRT methods have been the mainstay of smoking cessation treatment and will likely continue to serve a useful role, the next stage of progress will likely entail the development of tools designed with recognition of the importance of nonnicotine components of cigarette smoking. PMID- 16362404 TI - Response to novelty, social and self-control behaviors, in rats exposed to neonatal anoxia: modulatory effects of an enriched environment. AB - Perinatal asphyxia is a concern for public health and may promote subtle and long lasting neuropsychiatric disorders. In the present study, newborn Wistar rat pups underwent a repeated 20-min exposure to a 100% N2 atmosphere (or air) on postnatal days (pnd) 1, 3, 5, and 7. Half of the animals were housed during adolescence (pnd 21-35) in an enriched environment. The consequences on behavior were assessed throughout adolescence to adulthood. When scored for social performance, adolescent rats exposed to neonatal asphyxia exhibited exaggerated levels of anogenital sniffing behavior, which was normalized by enriched living. In air-exposed controls, enriched living increased the expression of affiliative and novelty-seeking behaviors, as compared to standard housing. However, this enrichment-induced behavioral plasticity was not found in rats neonatally exposed to asphyxia. At adulthood, levels of impulsivity and 5-HT2A receptors in the striatum were markedly increased in neonatal-asphyxia rats kept in standard housing conditions. Interestingly, impulsivity and receptor density were normalized by enriched rearing during adolescence. These findings indicate profound long-lasting behavioral alterations as a consequence of repeated neonatal asphyxia in rats. Beneficial effects of stimulation by an enriched environment during the still-plastic window of adolescence are suggested in these animals. PMID- 16362403 TI - A reliable model of intravenous MDMA self-administration in naive mice. AB - RATIONALE: MDMA is one of the most widely consumed recreational drugs in Europe. However, the mechanisms involved in the reinforcing properties of MDMA are still unclear. In this sense, the establishment of a reliable model of MDMA self administration in mice could represent an important approach to study the neuronal substrates associated with MDMA reward by using genetically modified mice. OBJECTIVES: To develop a reliable model of operant intravenous MDMA self administration in drug-naive mice. MATERIALS AND METHODS: Mice were trained to acquire intravenous self-administration of MDMA at different doses (0, 0.06, 0.125, 0.25, 0.5 and 1.0 mg/kg/infusion) on a FR1 schedule of reinforcement for 15 consecutive days. The motivational value of different doses of MDMA (0.125, 0.25 and 0.5 mg/kg/infusion) was then tested using a progressive ratio paradigm. Finally, [3H]-mazindol autoradiographic studies were carried out in order to quantitatively assess presynaptic dopamine transporter (DAT) binding sites in the striatum of mice trained to self-administer MDMA (0 and 1.0 mg/kg/infusion) during 15 days. RESULTS: The latency for discrimination between the active and inactive holes, as well as the number of animals acquiring stability criteria, varied as a function of the dose of MDMA. The mice responding for intermediate doses (0.125, 0.25 and 0.5 mg/kg/infusion) discriminated earlier than those responding for low (0.06 mg/kg/infusion) or high (1.0 mg/kg/infusion) doses. The percentage of animals achieving stability criteria increased with days of testing and was inversely proportional to the dose of MDMA. The breaking points achieved for doses of 0.125 and 0.25 mg/kg/infusion were significantly higher than for a dose of 0.5 mg/kg/infusion. No significant DAT neurotoxicity was observed in the striatum of animals self-administering MDMA at a dose of 1 mg/kg/infusion. CONCLUSIONS: The present results show that MDMA can be reliably self-administered by drug-naive mice. PMID- 16362405 TI - Effects of haloperidol, clozapine, and quetiapine on sensorimotor gating in a genetic model of reduced NMDA receptor function. AB - RATIONALE: Reduced N-methyl D-aspartate (NMDA) receptor function is hypothesized to contribute to the pathophysiology of schizophrenia. In order to model chronic and developmental NMDA receptor hypofunction, a mouse line was developed that expresses low levels of the NMDA R1 (NR1) subunit of the NMDA receptor. These mice show increased acoustic startle reactivity and deficits in prepulse inhibition (PPI) of acoustic startle. OBJECTIVES: The present study tested the hypothesis that these altered acoustic startle responses in the NR1 hypomorphic (NR1-/-) mice would be affected by antipsychotic drug treatment. METHODS: Mice were injected with drugs 30 min before assessment of acoustic startle responses with and without prepulse stimuli. RESULTS: Haloperidol (0.5 or 1.0 mg/kg) did not reduce the increased startle reactivity in the NR1-/- mice, but did increase PPI in both the mutant and wild type mice. Clozapine (3 mg/kg) and quetiapine (20 mg/kg) reduced startle magnitude and increased PPI in both the wild type and mutant mice. The antidepressant drug imipramine (10 and 20 mg/kg) had minimal effects on startle amplitude in NR1-/- or wild type mice. However, for the 20 mg/kg dose of imipramine, a significant increase in PPI was observed in the wild type animals, but not in the mutant mice. CONCLUSIONS: The results demonstrate that PPI can be increased in a mouse model of chronic NMDA receptor hypofunction by typical and atypical antipsychotic drugs. The similar effects of typical and atypical antipsychotic drugs to increase PPI in the wild type and mutant mice indicates that the assessment of behavior of the NR1 hypomorphic mice in the PPI paradigm offers no advantage over the wild type controls for identifying new clozapine-like drugs. PMID- 16362407 TI - Synthetic double-stranded RNA poly(I:C) as a potent peptide vaccine adjuvant: therapeutic activity against human cervical cancer in a rodent model. AB - Due to the inherent lack of immunogenicity of peptides, it is generally recognized that the strong inflammatory signals that are required to elicit specific responses against peptide-based therapeutic tumor vaccines may not be provided by the standard/conventional vaccine adjuvants. In this study, we have demonstrated dsRNA in the form of synthetic pI:C as a potent adjuvant to enhance the specific anti-tumor immune responses against a peptide-based vaccine. When complexed with an MHC I-restricted minimal peptide epitope derived from the HPV 16 E7 protein, the resulting pI:C/E7(49-57) molecular complex induced strong E7(49-57)-specific CTL responses that caused significant regressions of model human cervical cancer tumors pre-established in mice. In addition, although the proportion of DCs in tumor-bearing mice was significantly decreased when compared to that in naive mice, immunization with pI:C/E7(49-57 )restored the proportion of DCs in tumor-bearing mice. Double-stranded RNA may hold a great potential as an adjuvant to induce cellular immune responses for tumor immunotherapy. PMID- 16362408 TI - Strategies for immune intervention. PMID- 16362409 TI - Characterization of IL-2-activated TILs and their use in intrapericardial immunotherapy in malignant pericardial effusion. AB - Pericardial effusion (PE) and cardiac tamponade caused by malignant pericarditis are critical conditions in cancer patients, which still lack a recommended protocol for their long-term management. Percutaneous pericardiocentesis and simple drainage are commonly performed as the initial treatment. The aims of this study were to investigate the presence of cytotoxic T lymphocytes (CTLs) in malignant PE and to determine the clinical response to administering autologous tumor-infiltrating lymphocytes (TILs) into the pericardial cavity. Initially, we identified human lymphocyte antigen class-I-restricted and tumor-specific CTLs within the interleukin-2 (IL-2)-activated TILs in PEs from four patients, on the basis of interferon-gamma production and lactate dehydrogenase-release assays. Clinically we observed favorable responses to the pericardial transfer of IL-2 activated autologous TILs in four patients: one male with advanced esophageal cancer, one female with recurrent lung cancer and two females with recurrent breast cancer, respectively. Autologous TILs from PEs were expanded in vitro with IL-2, characterized for CD3, CD4 and CD8 markers, checked for contamination and then infused into the patient's pericardial space through a catheter. This was repeated biweekly. After treatment, there were no signs of recurrence of PE in either case, as determined by radiography, echocardiography and computed tomography. The only adverse effects seen were grade 1 fevers. These results suggested that intrapericardial cellular immunotherapy with autologous TILs could be a safe and effective treatment for controlling malignant pericarditis with associated cardiac tamponade, and that tumor-specific CTLs present in malignant PE might be important for tumor rejection. PMID- 16362406 TI - Action by and sensitivity to neuroactive steroids in menstrual cycle related CNS disorders. AB - Neuroactive steroids are a large group of substances having effect in the brain and on brain function. The steroids most studied are allopregnanolone (ALLO), tetrahydrodesoxycorticosterone (THDOC), pregnenolone sulfate (PS) dihydroepiandrosteronesulfate (DHEAS), and estradiol (E2). ALLO and THDOC are called gamma-aminobutyric acid (GABA) steroids as they are positive modulators of the GABAA receptor in a similar way as benzodiazepines, barbiturates, and alcohol. GABA steroids not only have similar behavioral effects as benzodiazepines and barbiturates but, possibly, also similar adverse effects as well. This review aims to elucidate the possible role that neuroactive steroids play in the development of mood disorders in women. One of the most clear-cut examples of the interaction between mood, neuroactive steroids, and the GABA system is premenstrual dysphoric disorder (PMDD), which is a cluster of negative mood symptoms occurring during the luteal phase of the menstrual cycle in 2-6% of reproductive women. Furthermore, certain women also experience adverse mood effects during sequential progestin addition to postmenopausal estrogen treatment, which is why the role of neuroactive steroids in postmenopausal women is also addressed in this review. PMID- 16362410 TI - Active hexose correlated compound enhances tumor surveillance through regulating both innate and adaptive immune responses. AB - Active hexose correlated compound (AHCC) is a mixture of polysaccharides, amino acids, lipids and minerals derived from cocultured mycelia of several species of Basidiomycete mushrooms. AHCC has been implicated to modulate immune functions and plays a protective role against infection. However, the potential role of AHCC in tumor immune surveillance is unknown. In this study, C57BL/6 mice were orally administered AHCC or water, followed by tumor cell inoculation. We showed that compared to pure water-treated mice, AHCC treatment significantly delayed tumor development after inoculation of either melanoma cell line B16F0 or lymphoma cell line EL4. Treatment with AHCC enhanced both Ag-specific activation and proliferation of CD4(+) and CD8(+) T cells, increased the number of tumor Ag specific CD8(+) T cells, and more importantly, increased the frequency of tumor Ag-specific IFN-gamma producing CD8(+) T cells. Interestingly, AHCC treatment also showed increased cell number of NK and gammadelta T cells, indicating the role of AHCC in activating these innate-like lymphocytes. In summary, our results demonstrate that AHCC can enhance tumor immune surveillance through regulating both innate and adaptive immune responses. PMID- 16362411 TI - Tumor escape mechanisms in prostate cancer. AB - Numerous immunotherapy trials have been carried out in prostate cancer (PC) patients, with induction of antigen-specific T cells in some cases. Despite this capability, limited success is seen in terms of tumor regression or survival. In this review, we discuss the evidence for tumor escape strategies that may contribute to vaccine failure in the setting of PC. These include defects in antigen presentation, production of immunosuppressive substances, induction of T cell death, T cell receptor dysfunction, and the presence of tolerogenic dendritic cells and regulatory T cells inside prostate tumors. It is clear that novel strategies aimed at preventing tumor escape, such as small molecular weight inhibitors of immunosuppressive molecules, adoptive transfer of TCR transgenic T cells, removal of Tregs, combined with anti-androgen therapy and prostate specific vaccines, need to be examined further in PC patients. PMID- 16362412 TI - Telomere length of in vivo expanded CD4(+)CD25 (+) regulatory T-cells is preserved in cancer patients. AB - PURPOSE: CD4(+)CD25(+) regulatory T-cells (Treg) are increased in the peripheral blood of cancer patients. It remains unclear whether this is due to redistribution or active proliferation. The latter would require the upregulation of telomerase activity, whose regulation also remains unknown for Treg. EXPERIMENTAL DESIGN: Treg and CD4(+)CD25(-) T-cells were isolated from peripheral blood of cancer patients (n=23) and healthy age-matched controls (n=17) and analyzed for their content of T-cell receptor excision circles (TREC) and for telomere length using flow-FISH, real-time PCR and Southern blotting. The in vitro regulation of telomerase of Treg was studied using PCR-ELISA in bulk cultures as well as in isolated proliferating and non-proliferating Treg. RESULTS: Treg isolated from peripheral blood of cancer patients exhibit significantly decreased levels of TREC when compared to Treg from healthy controls. Despite their in vivo proliferation, telomere length is not further shortened in Treg from cancer patients. Accordingly, telomerase activity of Treg was readily inducible in vitro. Notably, sorting of in vitro proliferating Treg revealed a significant telomere shortening in Treg with high-proliferative capacity. The latter are characterized by shortened telomeres despite high telomerase activity. CONCLUSIONS: Increased frequencies of Treg in peripheral blood of cancer patients are due to active proliferation rather than due to redistribution from other compartments (i.e., secondary lymphoid organs or bone marrow). In vivo expansion does not further shorten telomere length, probably due to induction of telomerase activity. In contrast, under conditions of strong in vitro stimulation telomerase induction seems to be insufficient to avoid progressive telomere shortening. PMID- 16362413 TI - D2-40 staining in sinonasal-type hemangiopericytoma--further evidence of distinction from conventional hemangiopericytoma and solitary fibrous tumor. AB - D2-40 is a monoclonal antibody, which reacts with a fixative-resistant epitope of lymphatic endothelium. Sinonasal-type hemangiopericytoma (SHP) and tumors of the (conventional) hemangiopericytoma/solitary fibrous tumor family (HP/SFT) are characterized by prominent vasculature. However, data concerning D2-40 labeling of these tumors are very sparse. In the present study, we investigated D2-40 staining in tissue specimens of 17 patients with SHP (male to female ratio of 2.4:1, median age of 63 years) and compared the immunolabeling with 20 cases of HP/SFT, including three SFT cases from nasal mucosa. D2-40 was detected in vascular channels of all SHP patients examined. By contrast, all cases of HP/SFT did not reveal any vascular channel being positive for D2-40, neither in the nasal cases nor in the remaining patients. This study presented for the first time data on D2-40 labeling in a series of SHP, HP/SFT, and supports the distinction of SHP from HP/SFT. PMID- 16362414 TI - Peritoneal metastasis inhibition by linoleic acid with activation of PPARgamma in human gastrointestinal cancer cells. AB - The effect on peritoneal metastasis of linoleic acid (LA) was examined using in vitro treatment of cancer cells and mouse peritoneal metastasis models. Firstly, cell growth of MKN28 human gastric cancer cells and Colo320 human colon cancer cells was suppressed by LA in a dose-dependent manner with increment of apoptosis. LA-induced growth inhibition was recovered by the exposure to antisense S-oligodeoxynucleotide for peroxisome proliferator-activated receptor gamma (PPARgamma) or 15-lipoxygenase-1, which converts LA to PPARgamma ligands. LA significantly inhibited invasion into type-IV collagen-coated membrane of MKN28 and Colo320 cells (p<0.05). BALB/c nu/nu mice inoculated with MKN28 and Colo320 cells into their peritoneal cavities were administrated with LA intraperitoneally (weekly, four times). The LA treatment significantly diminished the number of metastatic foci of both cells in the peritoneal cavity (p<0.05). Protein production in MKN28 and Colo320 cells treated with LA showed a decrease of epidermal growth factor receptor and an increase of Bax. These findings suggest that LA inhibits invasion and metastasis of human gastric and colon cancer cells by nondietary administration. PMID- 16362415 TI - Pan-selectin antagonism improves psoriasis manifestation in mice and man. AB - The selectin family of vascular cell adhesion molecules is comprised of structurally related carbohydrate binding proteins, which mediate the initial rolling of leukocytes on the activated vascular endothelium. Because this process is one of the crucial events in initiating and maintaining inflammation, selectins are proposed to be an attractive target for the development of new antiinflammatory therapeutics. Here, we demonstrate that the synthetic pan selectin antagonist bimosiamose is effective in pre-clinical models of psoriasis as well as in psoriatic patients. In vitro bimosiamose proved to be inhibitory to E- or P-selectin dependent lymphocyte adhesion under flow conditions. Using xenogeneic transplantation models, bimosiamose reduced disease severity as well as development of psoriatic plaques in symptomless psoriatic skin. The administration of bimosiamose in patients suffering from psoriasis resulted in a reduction of epidermal thickness and lymphocyte infiltration. The clinical improvement was statistically significant (P=0.02) as analyzed by comparison of psoriasis area and severity index before and after treatment. Assessment of safety parameters showed no abnormal findings. These data suggest that pan selectin antagonism may be a promising strategy for the treatment of psoriasis and other inflammatory diseases. PMID- 16362416 TI - The early vertebrate Danio rerio Mr 46000 mannose-6-phosphate receptor: biochemical and functional characterisation. AB - Mannose-6-phosphate receptors (MPRs) have been identified in a wide range of species from humans to invertebrates such as molluscs. A characteristic of all MPRs is their common property to recognize mannose-6-phosphate residues that are labelling lysosomal enzymes and to mediate their targeting to lysosomes in mammalian cells by the corresponding receptor proteins. We present here the analysis of full-length sequences for MPR 46 from zebrafish (Danio rerio) and its functional analysis. This is the first non-mammalian MPR 46 to be characterised. The amino acid sequences of the zebrafish MPR 46 displays 70% similarity to the human MPR 46 protein. In particular, all essential cysteine residues, the transmembrane domain as well as the cytoplasmic tail residues harbouring the signals for endocytosis and Golgi-localizing, gamma-ear-containing, ARF-binding protein (GGA)-mediated sorting at the trans-Golgi network, are highly conserved. The zebrafish MPR 46 has the arginine residue known to be essential for mannose-6 phosphate binding and other additional characteristic residues of the mannose-6 phosphate ligand-binding pocket. Like the mammalian MPR 46, zebrafish MPR 46 binds to the multimeric mannose-6-phosphate ligand phosphomannan and can rescue the missorting of lysosomal enzymes in mammalian MPR-deficient cells. The conserved C-terminal acidic dileucine motif (DxxLL) in the cytoplasmic domain of zebrafish MPR 46 essential for the interaction of the GGAs with the receptor domains interacts with the human GGA1-VHS domain. Interestingly, the serine residue suggested to regulate the interaction between the tail and the GGAs in a phosphorylation-dependent manner is substituted by a proline residue in fish. PMID- 16362417 TI - Regulation of cell adhesion in the Drosophila embryo by phosphorylation of the cadherin-catenin-complex. AB - Cell-culture studies indicate that tyrosine phosphorylation of the cadherin catenin-complex (CCC) is one of the post-translational mechanism regulating E cadherin-mediated cell adhesion. In this investigation, controlled application of a tyrosine phosphatase inhibitor (orthovanadate) and tyrosine kinase inhibitor (tyrphostin) to early Drosophila embryos, followed by biochemical assays and phenotypic analysis, has been utilized to address the mechanism by which tyrosine phosphorylation regulates E-cadherin-mediated cell adhesion in vivo. Our data suggest that, in the Drosophila embryo, beta-catenin (Drosophila homolog Armadillo) is the primary tyrosine-phosphorylated protein in the CCC. The increase in tyrosine phosphorylation correlates with a loss of epithelial integrity and adherens junctions in the ectoderm of early embryos. Late application of the phosphatase inhibitor does not have this effect, presumably because of the formation of septate junctions in late embryos. Co immunoprecipitation assays have demonstrated that tyrosine hyper-phosphorylation does not cause the dissociation of Drosophila (D)E-cadherin and alpha-catenin or Armadillo, suggesting that abrogation in adhesion is most likely attributable to the detachment of actin-associated proteins from the CCC. Finally, although the Drosophila epidermal growth factor receptor (EGFR), a receptor tyrosine kinase, is linked to the CCC and shows genetic interactions with DE-cadherin, we find that a constitutively active Drosophila EGFR construct does not cause any detectable changes in the level of tyrosine phosphorylation of Armadillo or destabilization of the CCC. PMID- 16362418 TI - Effect of two AMF life strategies on the tripartite symbiosis with Bradyrhizobium japonicum and soybean. AB - This study is the first in assessing the effect of soil disturbance on the contribution of arbuscular mycorrhizal fungi (AMF) with different life-history strategies to the tripartite symbiosis with soybeans and Bradyrhizobium japonicum (Kirchner) Jordan. We hypothesized that Gigaspora margarita Becker and Hall would be more affected by soil disturbance than Glomus clarum Nicol. and Schenck, and consequently, the tripartite symbiosis would develop more rapidly and lead to greater N(2) fixation in the presence of the latter. Soil pasteurization allowed the establishment of treatments with individual AMF species and soil disturbance enabled the development of contrasting root colonization potentials. In contrast, the colonization potential of B. japonicum was kept the same in all treatments. Soil disturbance significantly reduced root colonization by both AMF, with Gi. margarita being considerably more affected than G. clarum. Furthermore, the tripartite symbiosis progressed faster with G. clarum, and at 10 days after plant emergence, there was 30% more nodules when G. clarum was present compared to that when the bacterial symbiont alone was present. At flowering, the absence of soil disturbance stimulated N(2) fixation by 17% in mycorrhizal plants. However, this response was similar for both AMF. PMID- 16362419 TI - Binding of chromium(VI) to histones: implications for chromium(VI)-induced genotoxicity. AB - The first evidence has been obtained for Cr(VI) (chromate) binding to isolated calf thymus (CT) histones under physiological conditions (pH 7.4, Cl(-) concentration 152 mM, 310 K). No significant Cr(VI) binding under the same conditions was observed for other extracellular and intracellular proteins, including albumin, apo-transferrin and G-actin, as well as for CT DNA. The mode of Cr(VI) binding to histones was studied by vibrational, electronic and X-ray absorption (X-ray absorption near-edge structure and X-ray absorption fine structure) spectroscopies and molecular mechanics calculations. A proposed binding mechanism includes electrostatic interactions of CrO(4) (2-) with protonated Lys and Arg residues of histones, as well as the formation of hydrogen bonds with the protein backbone. Similarly, Cr(VI) can bind to nuclear localization signals (typically, Lys- and Arg-rich fragments) of other nuclear proteins. Selective binding of Cr(VI) to newly synthesized nuclear proteins (including histones) in the cytoplasm is likely to be responsible for the active transport of Cr(VI) into the nuclei of living cells. PMID- 16362420 TI - Scarred myocardium imposes additional burden on remote viable myocardium despite a reduction in the extent of area with late contrast MR enhancement. AB - Magnetic resonance imaging (MRI) can simultaneously detect and quantify myocardial dysfunction and shrinkage in contrast-enhanced areas postinfarction. This ability permits the investigation of our hypothesis that transformation of infracted myocardium to scarred tissue imposes additional burdens on peri infarcted and remote myocardium. Pigs (n = 8) were subjected to reperfused infarction. Gd-DOTA-enhanced inversion recovery gradient echo sequence (IR-GRE) imaging was performed 3 days and 8 weeks postinfarction. Global and regional left ventricular (LV) function was evaluated by cine MRI. Triphenyltetrazolium chloride (TTC) stain was used to delineate infarction while hematoxylin and eosin (H & E) and Masson's trichrome stains were used to characterize remodeled myocardium. Late contrast-enhanced MRIs showed a decrease in the extent of enhanced areas from 17 +/- 2% at 3 days to13 +/- 1% LV mass at 8 weeks. TTC infarction size was 12 +/- 1% LV mass. Cine MRIs showed expansion in dysfunctional area due to unfavorable remodeling, ischemia, or strain. Ejection fraction was reduced in association with increased end-diastolic and end-systolic volumes. Scarred myocardium contained collagen fibers and remodeled thick-walled vessels embedded in collagen. Sequential MRI showed greater LV dysfunction despite the shrinkage in extent of enhanced areas 2 months postinfarction. The integration of late enhancement and cine MRI incorporates anatomical and functional evaluation of remodeled hearts. PMID- 16362421 TI - High-resolution computed tomography of the lungs: the borderlands of normality. AB - High-resolution computed tomography (HRCT) is now widely used in the assessment of airways and diffuse lung disease. Considerable literature on pathologic correlation has increased the understanding of the signs of disease seen on HRCT. However, neither the significance of subtle individual signs nor the spectrum of HRCT appearances in healthy lungs is well documented. HRCT signs that cause diagnostic uncertainty and the spectrum of findings that exist between definite normality and definite abnormality are discussed. PMID- 16362422 TI - Comparative study of promoters for the production of polyhydroxyalkanoates in recombinant strains of Wautersia eutropha. AB - Recombinant strains of Wautersia eutropha expressing an artificial polyhydroxyalkanoate (PHA) biosynthesis operon under the control of different native promoters linked to polyhydroxybutyrate (PHB) (P(phb)), acetoin (P(acoE), P(acoD), and P(acoX)) or pyruvate (P(pdhE)) metabolism were constructed and tested. The promoters were representative either of the enterobacterial sigma70 (P(phb), P(acoE), and P(pdhE))- or sigma54 (P(acoD) and P(acoX))-dependent promoters. To obtain polymers consisting of C4-C12 monomer units, an artificial operon consisting of the PHA synthase gene from Pseudomonas sp. 61-3 (phaC1 (Ps)) tandemly linked to the W. eutropha genes encoding beta-ketothiolase (phbA (We)) and nicotinamide adenine dinucleotide phosphate dependent acetoacetyl-coenzyme A (CoA) reductase (phbB (We)) was constructed. All recombinant strains produced PHA, indicating that the PHA biosynthesis genes were expressed under the control of the different promoters. Cell growth and PHA synthesis on MS medium complemented with gluconate or octanoate, and different concentrations of acetoin (0, 0.15, and 0.3%) clearly differed among the recombinant strains. While the P(acoD) and P(acoX) promoters mediated only low PHA yields (<1%) in the presence of the inducer acetoin, the remaining promoters-independent of the addition of acetoin-resulted in the production of PHA polymers with high 3HB fractions (90 100 mol%) and with high 3HO contents (70-86 mol%) from gluconate and octanoate, respectively. Interestingly, on octanoate-MS medium with 0.15% acetoin, the P(acoE) promoter mediated the synthesis of PHA with a relatively high 3HB fraction (48 mol%). While PHAs with high 3HB contents were obtained, the overall PHA product yields were low (<10%); thus, their potential application for further commercial exploitation appears limited. PMID- 16362423 TI - Subacute posteromedial impingement of the ankle in athletes: MR imaging evaluation and ultrasound guided therapy. AB - OBJECTIVE: To describe the use of MR imaging and efficacy of ultrasound-guided steroid injection in the diagnosis and management of athletes with clinical posteromedial impingement of the ankle. DESIGN AND PATIENTS: A retrospective analysis of imaging findings on MR was undertaken in nine elite athletes with clinical posteromedial ankle impingement. MR studies from six professional athletes with posterolateral pain were also reviewed as an imaging control group. The two reviewing radiologists were blinded to the clinical details and the proportion of control and study subjects. The nine study athletes also underwent diagnostic ultrasound and ultrasound-guided injection of steroid and anaesthetic into the posteromedial capsular abnormality. Follow-up was by telephone interview. RESULTS: Posteromedial capsular thickening was seen only in athletes with posteromedial impingement (7/9). Posteromedial synovitis was present in all athletes with posteromedial impingement; however, posterior and posterolateral synovitis was also seen in these athletes. Mild posteromedial synovitis was present in two control athletes. Ultrasound identified abnormal posteromedial soft tissue thickening deep to tibialis posterior between the medial malleolus and talus in all nine athletes. After injection all athletes returned to their previous level of sport, with eight of the nine not experiencing any residual or recurrent symptoms. CONCLUSION: If MR imaging excludes significant coexistent abnormality, ultrasound can localise posteromedial soft tissue abnormality and guide injection therapy, allowing return to athletic activity without surgical intervention. PMID- 16362425 TI - Bilateral intertrochanteric fractures after surgical treatment of bilateral femoral neck fractures secondary to hypocalcemic convulsions with chronic renal failure: a case report and review of the literature. AB - There have been no reports of bilateral femoral neck fractures followed by bilateral femoral intertrochanteric fractures. A 28-year-old man presenting with bilateral femoral neck fractures due to generalized convulsions was diagnosed with chronic renal failure and treated with percutaneous screw osteosynthesis and hemodialysis. During rehabilitation, the patient again had convulsions resulting in bilateral femoral intertrochanteric fractures 2 months after the operation. He was treated with a compression hip screw on the right side and conservatively on the left side. This is the first well-documented report on bilateral femoral neck fractures followed by bilateral femoral intertrochanteric fractures. PMID- 16362426 TI - Ingrowth of osteochondral grafts under the influence of growth factors: 6-month results of an animal study. AB - INTRODUCTION: In the age of growth factors and gene therapy, the induction of cartilage healing remains an unsolved problem. Even in autologous grafting, one of the preferred methods of treatment for focal osteochondral lesions, chondral integration remains difficult. This study aims to define a possible positive influence of growth factor augmentation on the ingrowth of these transplants. MATERIALS AND METHODS: In an ovine model, questions regarding the healing of osteochondral transplants under the influence of two different growth factors were to be addressed. Two osteochondral autologous transplantations (OAT), one in the weight-bearing surface of each femoral condyle, were performed on the ovine knee using the standard operative protocol. One of the grafts was bathed in augmented PBS containing 50 microg bFGF or bone morphogenetic protein (BMP)-2 directly prior to implantation, while the other condyle served as the control. Two groups, consisting of eight sheep each, were evaluated for each growth factor after 6 months. RESULTS: During the evaluation of all the specimens, neither osteophytes nor synovial changes were observed. The mechanical consistency of the cartilaginous tissue began to reach a level equivalent to the surrounding tissue at 6 months, independent of the use of growth factor. Macroscopically, the superficial border of the transplanted osteochondral plug could easily be outlined in all groups, while the cartilage interface of the bFGF specimens was determined to be less demarcated than the BMP augmented plugs or the controls. Radiographically, a solid osteointegration of the graft could be documented at 6 months in the native and augmented groups. In contrast, integration of the chondral surface of the OAT was not seen macro- or microscopically in any specimen, even though cartilage surfaces remained viable. A firm physical interdigitation of the reconstructed joint surface could not be demonstrated in either of the two augmented groups or the control population. The augmentation with bFGF and BMP-2 stimulated the osseous ingrowth and seems to expedite the remodelling process, but was not able to improve chondral healing. CONCLUSION: The lack of integration of the cartilaginous portion of the transplanted plugs into the reconstructed joint surface, even following the augmentation with bFGF and BMP-2, does not bode well for the long-term survival of the joint itself. PMID- 16362424 TI - Genome-wide analysis of differentially expressed genes from Penicillium chrysogenum grown with a repressing or a non-repressing carbon source. AB - Penicillium chrysogenum is an economically important ascomycete used as industrial producer of penicillin. However, with the exception of penicillin biosynthesis genes, little attention has been paid to the genetics of other aspects of the metabolism of this fungus. In this article we describe the first attempt of systematic analysis of expressed genes in P. chrysogenum, using a suppression subtractive hybridization approach to clone and identify sequences of genes differentially expressed in media with glucose or lactose as carbon source (penicillin-repressing or non-repressing conditions). A total of 167 clones were analysed, 95 from the glucose condition and 72 from the lactose condition. Genes differentially expressed in the glucose condition encode mainly proteins involved in the mitochondrial electron transport chain and primary metabolism. Genes expressed differentially in lactose-containing medium include genes for secondary metabolism (pcbC, isopenicillin N synthase), different hydrolases and a gene encoding a putative hexose transporter or sensor. The results provided information on how the metabolism of this fungus adapts to different carbon sources. The expression patterns of some of the genes support the hypothesis that glucose induces higher rates of respiration in P. chrysogenum while repressing secondary metabolism. PMID- 16362427 TI - Xenomict energy in cold solids in space. AB - Minerals on earth whose crystalline order has been reduced by radioactive decay of contained atoms are termed "metamict." They are rare and few because in most crystalline solids, atoms and vacancies are relatively mobile at terrestrial temperatures, and radiation damage tends to be self-annealing. This is not the case in the extreme cold of deep space. Below roughly 100 K, reduced vacancy mobility allows cosmic ray and solar wind induced lattice defects to endure and accumulate for eons, reaching energy densities of up to MJ kg(-1) in some materials. We examine the possible effects of the release of energy stored in cold deep-space materials when solid-state defects recombine upon warming due to impacts, gravitational infall, or perihelion. Dimensional analysis suggests energetic defect recombination in radiation-damaged "xenomict" solids in comets, and planetesimals may, in some circumstances, raise internal temperatures enough to melt ice and volatilize frozen gases. We speculate that this may account for some cometary outbursts and Deep Impact experiment results. Calorimetric experiments on appropriately irradiated natural and synthetic materials are needed to further quantify these mechanisms. PMID- 16362428 TI - Pramipexole protects against H2O2-induced PC12 cell death. AB - Pramipexole, a novel non-ergot dopamine (DA) agonist, has been successfully applied to the treatment of Parkinson's disease (PD). Although the specific cause of PD remains unknown, recent studies have provided evidence that oxidative stress plays a role in the parthenogenesis of the disease. In the present study, we examined the effect of pramipexole on hydrogen peroxide (H2O2, 100 microM) induced PC12 cell death, and the intracellular mechanism of this effect. The 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay revealed that pretreatment of PC12 cells with pramipexole (1-100 microM) resulted in significant protection against H2O2-induced cell death in a concentration dependent manner. The protective effect of pramipexole was not affected by pretreatment with the DA receptor antagonists sulpiride, spiperone or domperidone, suggesting that the effect of pramipexole is not mediated by DA receptors. In PC12 cells, pramipexole inhibited H2O2-induced lactate dehydrogenase (LDH) leakage, as well as H2O2-induced cytochrome c release and caspase-3 activation with the resultant apoptosis. It was also observed in PC12 cells that H2O2 stimulated phosphorylation of mitogen-activated protein (MAP) kinases, i.e., extracellular signal-regulated kinase1/2 (ERK1/2), c-Jun NH2 terminal kinase (JNK) and p38 MAP kinase. Pramipexole inhibited H2O2-induced JNK and p38 MAP kinase, but not ERK1/2 phosphorylation. Furthermore, in these cells experiments with a fluorescent probe, 2-[6-(4'-amino)phenoxy-3H-xanthen-3-on-9 yl]benzoic acid, revealed that pramipexole, the JNK inhibitor SP600125 and the p38 MAP kinase inhibitor SB203580 inhibited the generation of H2O2-induced reactive oxygen species. Caspase inhibitors Z-DEVD-FMK and Z-IETD-FMK, as well as SP600125 and SB203580, inhibited H2O2-induced PC12 cell death to a similar extent as pramipexole. These results suggest that pramipexole exerts a protective effect against oxidative stress-induced PC12 cell death in part through an inhibition of JNK and p38 MAP kinase. PMID- 16362429 TI - Allosteric site in M2 acetylcholine receptors: evidence for a major conformational change upon binding of an orthosteric agonist instead of an antagonist. AB - Muscarinic acetylcholine receptors contain two distinct ligand binding sites, i.e. the orthosteric site for acetylcholine and other conventional ligands, and an allosteric site located at the entrance of the ligand binding pocket. We used a set of allosteric agents to probe whether muscarinic M2 receptors whose orthosteric site is occupied by an agonist still reveal the common allosteric site that has been identified in M2 receptors being occupied by an orthosteric antagonist (N-methylscopolamine, NMS). Equilibrium and dissociation binding experiments were carried out in porcine heart homogenates using either the agonist [3H]oxotremorine M ([3H]OxoM) or the antagonist [3H]NMS. The affinities of the allosteric agents were determined for the radioligand-occupied receptor states and, additionally, for the radioligand-free (ground state) M2 receptor. The archetypal agent W84 (hexane-1,6-bis[dimethyl-3'-phthalimidopropyl-ammonium bromide] and its bispyridinio middle chain analogue WDuo3 (1,3-bis[4 (phthalimidomethoxyimino-methyl)-pyridinium-1-yl]propane dibromide) had a clearly lower affinity for [3H]OxoM-liganded receptors compared with [3H]NMS-liganded and ground state receptors. In contrast, a derivative resembling only one half of W84 had equal affinities for both radioligand-occupied receptor states. Also, the agents gallamine and obidoxime did not discriminate between [3H]OxoM- and [3H]NMS occupied receptors. The allosteric antagonistic tool obidoxime inhibited WDuo3 action in [3H]OxoM-liganded receptors with the same potency as in [3H]NMS liganded receptors. We conclude that the common allosteric site is still present in OxoM-liganded M2 receptors, but its spatial conformation is considerably altered compared with NMS-liganded receptors. PMID- 16362430 TI - Volumetric assessment of myocardial viability in rats using 3D double contrast enhanced T1 and T2-weighted MRI. AB - OBJECTIVE: Volumetric evaluation of the myocardial viability post-infarction in rats using 3D in vivo MR imaging at 7 T using injection of an extracellular paramagnetic contrast agent and intravascular superparamagnetic iron oxide nanoparticles in the same imaging session. MATERIALS AND METHODS: Five hours after induction of permanent myocardial infarction in rats (n=6), 3D in vivo T1- and T2-weighted MR Imaging was performed prior to and after Gd-DOTA injection (0.2 mmol/kg) and prior to and after nanoparticle injection (5 mg Fe/kg) to assess infarct size and myocardial viability. RESULTS: 3D MR Imaging using a successive contrast agent injection showed a difference of infarct size after Gd DOTA injection on T1-weighted images compared to the one measured on T2-weighted images after Gd-DOTA and nanoparticle injection. CONCLUSION: The use of 3D T1- and T2-weighted MR Imaging using a double contrast agents protocol made possible the accurate characterization of myocardial infarction volume and allowed the detection of myocardial viability post-infarction in rats. PMID- 16362431 TI - Roots of the maxillary first and second molars in horizontal relation to alveolar cortical plates and maxillary sinus: computed tomography assessment for infection spread. AB - The purposes of this study were to investigate the horizontal relationship of the roots of maxillary molars with the cortical plates and the maxillary sinus and to investigate the influence of these relationships on the spread of odontogenic infection. Computed tomography images of 120 control subjects and 49 patients with infection originating in the maxillary first or second molar were investigated. In the control group, more than 60% of the first molar roots contacted both palatal and buccal cortical plates (type A), while such contact was not seen in more than 60% of second molars. The floor of maxillary sinus was most frequently observed at the level between the bifurcation and apices of roots in both first and second molars. In patients with infection, cortical changes were more frequently seen on the buccal side than on the palatal side, and 80% of patients with buccal cortical change showed the position in which the buccal roots were close to the buccal cortical plate. Mucosal thickening of the maxillary sinus was found in 87.8%. The buccopalatal spread of odontogenic infection originating in the maxillary first and second molars was influenced by the horizontal root position in relation to the cortices. PMID- 16362442 TI - Wegener's granulomatosis: clinical and laboratory results of a university hospital study of 20 patients from Turkey. AB - The aim of this study was to evaluate the clinical and laboratory features, the treatment approaches, and the long-term outcome of patients with Wegener's granulomatosis (WG) who were followed up in our hospital. The hospital files of the patients with the diagnosis of WG who were followed up between the years 1985 and 2003 in Hacettepe University Hospital were retrospectively evaluated. Male/female ratio was 12:8. The mean age was 39 years (range 20-65 years). Constitutional symptoms and upper and lower airway involvement were seen in 95% of all patients. Renal and musculoskeletal symptoms were seen in 90 and 80% of the patients, respectively. Five patients were treated with oral monotherapy (three with methylprednisolone and two with cyclophosphamide). Three patients were given a combination of orally administered cyclophosphamide and methylprednisolone. Ten patients were treated with pulse cyclophosphamide and methylprednisolone combination together with oral alternate-day methylprednisolone therapy. The remaining two resistant patients were treated with pulse cyclophosphamide, methylprednisolone, and intravenous immunoglobulin combination together with oral alternate-day methylprednisolone. Four patients died because of the disease activity. Intravenous pulse therapies with oral, alternate-day methylprednisolone were well tolerated. Sixteen patients experienced long-term remission after immunosuppressive treatment. Eleven patients have been asymptomatic for more than 12 months. In five patients, residual symptoms persisted: constitutional symptoms and renal and respiratory tract symptoms in varying combinations. The demographic and laboratory findings in this trial were similar with those of the previous results. Alternate-day glucocorticoids plus cyctotoxic drugs may be beneficial in patients with WG. PMID- 16362443 TI - Severity of rheumatoid arthritis: the SEVERA study. AB - This study aims to assess the severity of rheumatoid arthritis (RA) in rheumatology practice in our population. All outpatients and inpatients with RA seen by registered rheumatologists over a 1-year period were included. Severity was measured using the Larsen score for hands and wrists and the Modified Health Assessment Questionnaire (M-HAQ). Two hundred ninety-eight RA cases were included. Mean age was 51.5 years. Among them, 261 (87.6%) were females. Disease duration was less than a year in 26 subjects (8.7%) and 10 years and above in 108 (36.2%) with a mean of 8.9. There were 220 (73.8%) subjects who had M-HAQ score <1. In 61 (20.5%) subjects, M-HAQ score was > or =1 and <2, and 17 (5.7%) had M- HAQ score > or =2. In relation with disease duration, M-HAQ starts with an average (SD) value of 0.7 (0.6) during the first year, decreases to 0.4 (0.4) at 5-year disease duration and increases after 10 years of disease progression to an average of 0.9 (0.8). Mean (SD) Larsen score was 51.9 (29.5) and median was 45. A total of 25% had a Larsen score > or =50% of maximum. Larsen score increased significantly (p<0.0001) with disease duration, starting at an average (SD) of 36.1 (14.9) during the first year, rising to 42.5 (15.8) around 5 years and reaching 73.9 (36.9) after 10 years. RA severity in our practice is comparable to that reported in Western populations in terms of radiological damage; however, functional status differs, possibly reflecting cultural differences. PMID- 16362444 TI - Juvenile rheumatoid arthritis and bronchiolitis obliterans organized pneumonia. AB - Diverse pleuropulmonary manifestations, including pleural effusion, rheumatoid nodulosis, fibrosis, obliterans brochiolitis, bronchiectasias, vasculitis, drug induced lung disease, and obliterans bronchiolitis with organized pneumonia, have been described in patients with rheumatoid arthritis (RA). Bronchiolitis obliterans organized pneumonia (BOOP) is an uncommon condition described in patients with RA but not in juvenile RA (JRA). We described a patient with JRA who developed a BOOP. PMID- 16362445 TI - Systemic lupus erythematosus complicated by dilated cardiomyopathy and severe heart failure. AB - Systemic lupus erythematosus (SLE) is the prototype autoimmune disorder, one that is known for its many, diverse modes of presentation. In this paper, we present a further unusual presentation of SLE, that of acute onset, severe heart failure secondary to dilated cardiomyopathy. Only a few similar cases have been reported in the literature. PMID- 16362446 TI - Successful treatment of pediatric systemic polyarteritis nodosa with cholestatic hepatitis. AB - Polyarteritis nodosa (PAN) is a rare vasculitis in children. We report a case of 5-year-old boy with systemic PAN and cholestatic hepatitis. He had fever, abdominal pain, and gastrointestinal bleeding. Angiography revealed multiple microaneurysms in the renal, hepatic, and superior mesenteric arteries. Clinical manifestations improved slowly after immunosuppressive therapy, but liver enzyme and bilirubin levels elevated gradually. Liver biopsy findings revealed marked centrizonal canalicular cholestasis, bile duct damage, and intact hepatocyte, but there was no evidence of viral hepatitis or vasculitis. Levels of liver enzymes and bilirubin improved after two cycles of cyclophosphamide therapy. We thought that the possible etiology of elevated liver enzyme and bilirubin levels might be a manifestation of PAN. PMID- 16362447 TI - Seronegative Sjogren syndrome with asymptomatic autoimmune sclerosing pancreatitis. AB - We report two elderly patients with seronegative Sjogren syndrome who showed benign swelling of the pancreas on computed tomography. Immunostaining of the biopsied lip tissue or serum examination confirmed an increase in production of IgG4, leading to a diagnosis of autoimmune sclerosing pancreatitis (ASP) as a cause of the asymptomatic swelling of the pancreas. Sicca symptoms and ASP spontaneously improved in one patient, and the other responded well to oral prednisolone. Seronegative Sjogren syndrome and ASP can concurrently occur as a clinical manifestation of the IgG4-related systemic disorder, particularly in elderly subjects, and, in such a case, corticosteroid may be a potent therapeutic option. PMID- 16362448 TI - Clinical significance of IL-18, IL-15, IL-12 and TNF-alpha measurement in rheumatoid arthritis. AB - The aim of the study was to evaluate the clinical significance of serum (S) and synovial fluid (SF) interleukin (IL)-18, IL-15, IL-12 and the tumor necrosis factor alpha (TNF-alpha) measurements in relation to laboratory and clinical measures of disease activity of patients with active rheumatoid arthritis (RA). Sixty-four patients with RA and 25 patients with osteoarthritis (OA) were included in this study. RA activity was determined using the Disease Activity Score (DAS) 28 index. Concentrations of IL-18, IL-15, IL-12 and TNF-alpha were measured by ELISA. Serum C-reactive protein (CRP) levels were also determined. Cross-sectional correlations between S and SF levels of cytokines and values of DAS 28 index were calculated. The results have shown that IL-18, IL-15, IL-12 and TNF-alpha levels in S and SF of patients with RA were significantly higher than the levels obtain from patients with OA (p<0.01). Significantly higher levels of IL-18, IL-15 and TNF-alpha were found in the SF compared to the S of patients with RA (p<0.01). Significantly higher S and SF levels of all four cytokines and serum CRP values were found in RA patients with high disease activity (DAS 28>5.1) compared to those with mild (DAS 28>3.2) and low disease activity (DAS 28>2.6) (p<0.01). Serum and SF concentrations of all four cytokines positively correlated with DAS 28 index values, i.e., disease activity. A poor correlation was found for S and SF IL-12 whereas the highest coefficient of correlation was found for SF IL-18 (r=0.879, p<0.01), and SF TNF-alpha (r=0.827, p<0.01) and disease activity in this study. Strong correlation was found between SF TNF-alpha and SF IL-18 levels (r=0.732, p<0.01). In conclusion, SF IL-18 and TNF-alpha levels in RA patients are good indicators of disease activity. The results obtained support the use of the DAS in clinical practice as a reliable method in assessing disease activity in RA patients. PMID- 16362449 TI - Syringomyelia, neuropathic arthropathy and rheumatoid arthritis as diagnostic dilemmas in two different cases: confounding factor and true coexistence. AB - Two patients with neuropathic arthropathy (NA) of hands and wrists due to cervical syringomyelia are described. Articular deformities resembling rheumatoid arthritis (RA) were present in both cases. RA was misdiagnosed in the first case, leading to incorrect treatment with iatrogenic consequences, and there was real coexistence of RA and NA in the second one. Physicians must be aware of this rare condition in the differential diagnosis of wrist and hand deformities. PMID- 16362452 TI - Biochemical and physiological properties of a novel series of long-acting insulin analogs obtained by acylation with cholic acid derivatives. AB - PURPOSE: This study was conducted to assess the suitability of insulin analogs acylated by various cholic acid derivatives for use as basal insulin, and to test the most promising of these, LysB29(Nepsilon-lithocholyl-gamma-Glu) des(B30) human insulin (NN344) in pigs. METHODS: Circular dichroism spectroscopy and size exclusion chromatography were used to explore the physicochemical properties of the analogs, and affinities for albumin and insulin receptors were determined. After subcutaneous injection in pigs, disappearance half-times were measured, and the plasma profile and glucose-lowering effect in a euglycemic clamp were assessed for NN344. RESULTS: NN344 showed glucose-lowering activity lasting more than 24 h. Glucose infusion rate was essentially constant from 5 to 19 h after injection. NN344 seemed to be a dodecamer in the presence of zinc ions and phenol. Without phenol, the apparent molecular mass was >5000 kDa. Formation of such a self-assembly at the site of s.c. injection and its subsequent slow decomposition might explain the long duration of action of NN344. A measurable affinity for albumin of the lithocholic acid ligand may also contribute to the prolonged action. CONCLUSIONS: NN344 is a candidate for a neutral soluble basal insulin that might offer people with diabetes a prolonged duration, smooth, and predictable basal insulin supplement. PMID- 16362453 TI - Transdermal delivery of heparin using pulsed current iontophoresis. AB - PURPOSE: In clinical practice heparin has to be administered by injection with obvious disadvantages; thus, transdermal delivery by electrically assisted methods have been studied. In this study we evaluated the efficacy of a Food and Drug Administration-approved pulsed current iontophoresis system in delivering heparin through living rat skin. METHODS: Fluorescent and radioactive heparin as well as a commercial heparin preparation were delivered through rat skin via a pulsed current iontophoresis system. RESULTS: Pulsed current iontophoresis allowed fluorescent heparin to cross the stratum corneum localizing in epidermis and dermis. Unfractionated, high-, and low molecular weight fraction pools, obtained by fractionating [35S]-unfractionated heparin on a molecular weight sieve, were then separately tested. Pulsed current iontophoresis elicited the transdermal delivery of low molecular weight heparin, but not that of high molecular weight heparin. Finally, pulsed current iontophoresis of an unfractionated pharmaceutical heparin preparation significantly decreased plasmatic factor Xa activity. CONCLUSIONS: We hypothesize that this technique could be used to administer low molecular weight heparin in a cost-efficient and safe manner without the need for syringes and needles. PMID- 16362454 TI - Effect of zoledronate on bone quality in the treatment of aseptic loosening of hip arthroplasty in the dog. AB - Periprosthetic bone loss, which is a direct cause of aseptic loosening in total hip arthroplasty (THA), can be suppressed by bisphosphonates. It is unknown how the quality of this bone is affected in the presence of both wear debris (from implant) and bisphosphonates. The objective of this study was to evaluate the effect of zoledronate (ZLN) on bone quality in the presence of wear debris [polyethylene (PE) particles] in a canine model of uncemented THA. Thirty dogs underwent THA, and aseptic loosening was induced via implantation of PE particles packed into the femoral component. For 26 weeks until sacrifice, two groups (each n = 10) received weekly injections of ZLN (low dose 2 mug/kg, high dose 10 mug/kg) and the third group (control) received saline. Histological and radiographic examinations were performed to evaluate the degree of implant reaction. Histomorphometry (static/dynamic) was performed to evaluate bone turnover. Back-scattered electron imaging was used to quantify the newly formed bone and to evaluate the mineralization distribution. Density fractionation and X ray diffraction were used to evaluate mineral properties, while four-point bending was used to determine mechanical properties. A dose-dependent presence of newly formed subperiosteal bone was found, which appeared to be less mineralized than the adjacent cortical bone. The high-dose ZLN group showed decreased cortical porosity and turnover and increased mineralization profile, failure strength, and modulus. We conclude that ZLN affects some of the material properties of cortical bone and allows the newly formed subperiosteal bone to remain and therefore affect the overall quality of the bone. PMID- 16362455 TI - Effects of the Cdx-2 polymorphism of the vitamin D receptor gene and lifestyle factors on bone mineral density in a representative sample of Japanese women: the Japanese Population-based Osteoporosis (JPOS) Study. AB - Using a large-scale representative sample of the Japanese female population, we examined the effects of a single nucleotide polymorphism within a binding site of Cdx-2 in the promoter region of the vitamin D receptor gene on bone mineral density (BMD), and the interactions between this polymorphism and lifestyle factors on BMD. Fifty women were randomly selected from each of the 5-year age stratified populations (15-79 years) in each of three chosen municipalities as a part of the Japanese Population-based Osteoporosis Study. BMD at the lumbar spine, hip, and distal forearm was measured using dual-energy X-ray absorptiometry at baseline and again in a follow-up study conducted 3 years later. Information on lifestyle factors was collected in a questionnaire and followed up in interviews. The G-to-A polymorphism within the Cdx-2 binding site was determined by a TaqMan allelic discrimination assay. At baseline, 1,340 women were analyzed. The baseline BMD in the ultradistal forearm in premenopausal women with the GG genotype was significantly lower than in those with other genotypes. There was no association between the Cdx-2 genotype and the change in BMD at any of the skeletal sites. We found significant associations between daily milk consumption and baseline BMD at some skeletal sites but only in subjects with the GG genotype. In conclusion, the Cdx-2 polymorphism alone did not have a substantial effect on BMD in Japanese women. However, this polymorphism might have some effect in women with low calcium intake. PMID- 16362456 TI - Bioavailability of calcium: comparison of calcium carbonate and milk and the effect of vitamin D, age, and sex using 24-hour urine calcium as a method. AB - The aim of the present study was to compare the bioavailability of calcium from calcium carbonate and milk and to investigate if 1,200 IU of cholecalciferol a day increased intestinal absorption of calcium. Both young women and a group of older persons of both sexes were included to study the influence of age and sex. In total, 53 healthy women and men were included: a group of 23 younger women (median age 30) and an older group of 15 women and 15 men (median age 66). The study period was 4 weeks; each participant completed four treatment regimens randomly: CaCO(3), CaCO(3 )+ 1,200 IU of cholecalciferol, milk, and placebo. All regimens were distributed three times a day and consisted of 1,200 mg of elementary calcium. The 24-hour urine calcium excretion was used as a method. Total urinary calcium excretion rates (mmol/day) were as follows (mean +/- SD): placebo 4.41 +/- 2.17, milk 5.17 +/- 2.33, CaCO(3) 5.83 +/- 2.03, and CaCO(3 )+ D 6.06 +/- 2.46. All regimens compared to placebo were significant. Addition of cholecalciferol to the CaCO(3) regimen increased calcium excretion but insignificantly: 0.27 +/- 2.84 mmol/day. The increase in calcium excretion during the milk regimen was significant only for the old group: 0.96 vs. 0.28 mmol/day. No other difference was found according to age and sex. The bioavailability of calcium carbonate and milk was demonstrated. Additional cholecalciferol (1,200 IU) to individuals in positive calcium balance with serum 25(OH)D levels >50 nmol/L only marginally increased calcium absorption in a short-term intervention. PMID- 16362457 TI - Genetic and environmental correlations between age at menarche and bone mineral density at different skeletal sites. AB - Low bone mineral density (BMD) is an important risk factor for osteoporotic fractures. Though previous studies have demonstrated that age at menarche (AAM) is phenotypically associated with BMD, the contributions of genetic and environmental factors to this association remain unknown. In this study, using variance decomposition analyses, we provided an accurate estimation of the genetic and environmental correlations between AAM and BMD in 2,667 Caucasian women from 512 pedigrees. After adjustment for significant covariates, we detected significant genetic correlations between AAM and BMD at the lumbar spine, femoral neck, and ultradistal radius (rho(G) = -0.1316, -0.1417, and 0.1137, respectively; all P < 0.01). However, all environmental correlations between AAM and BMD were nonsignificant (P > 0.05). We also generated a principal component factor for BMD (PC_BMD) and evaluated the relationship between this factor and AAM. The genetic and environmental correlations between PC_BMD and AAM (rho(P) = -0.0847, P < 0.001; rho(G) = -0.1737, P < 0.01; rho(E) = -0.0348, P > 0.05) were consistent with the results of BMD at the three skeletal sites and AAM. Our results confirmed the significant phenotypic association between BMD and AAM and for the first time suggested that this association is mainly attributable to shared genetic, rather than environmental, factors. PMID- 16362458 TI - BMP2 gene therapy on the repair of bone defects of aged rats. AB - Age-related decline in the number of mesenchymal stem cells (MSCs) and their reduced capability to differentiate osteogenically, along with diminished availability of growth factors, may be major factors accounting for reduced bone formation in the aging mammalian body. In the first part of the study, we compared the number of MSCs in bone marrow (BM) and the content of bone morphogenetic protein 2 (BMP2) in cortical bone tissue in juvenile, adult, and aged (1, 9, and 24 months, respectively) male rats. To assay the influence of aging on osteogenic differentiation ability, MSCs from the three age groups were transduced with the BMP2 gene. Following gene transduction, the production of BMP2 in culture media, expression of osteogenic proteins (e.g., alkaline phosphatase, type Ialpha1 collagen, osteopontin, and bone sialoprotein), as well as ectopic bone formation in athymic mice were compared. Results showed that the number of MSCs in BM as well as the content of BMP2 in cortical bone tissue decreased with age, but no significant differences between the three age groups were found with regard to production of BMP2 or capability of BMP2 gene-modified MSCs to differentiate osteogenically. The second part of the study applied BMP2 gene-modified autologous MSCs/beta-tricalcium phosphate for repair of bone defects in aged rats with positive results. Our data indicate that the osteogenic potential of MSCs of aged rats can be restored following BMP2 gene transduction and that this technique may be a useful approach in the future planning of gene therapy for age-related osteoporotic fractures. PMID- 16362459 TI - The receptor activator of nuclear factor-kappaB ligand inhibitor osteoprotegerin is a bone-protective agent in a rat model of chronic renal insufficiency and hyperparathyroidism. AB - Osteoprotegerin (OPG) acts by neutralizing the receptor activator of nuclear factor-kappaB ligand (RANKL), the primary mediator of osteoclast differentiation, function, and survival. We examined whether OPG could affect the bone loss associated with chronic kidney disease (CKD) in a rodent model of CKD and secondary hyperparathyroidism (SHPT). SHPT was induced in rats by 5/6 nephrectomy (5/6 Nx) and a 1.2% P/0.6% Ca(2+) diet. Starting 1 week after 5/6 Nx, rats were treated with vehicle (veh) or OPG-Fc (3 mg/kg, intravenously) every 2 weeks for 9 weeks. At baseline, 3, 6, and 9 weeks, blood was taken and bone mineral density (BMD) and bone mineral content (BMC) were assessed by dual-energy X-ray absorptiometry. Serum parathyroid hormone (sPTH) levels reached 912 pg/ml in 5/6 Nx rats vs. 97 pg/ml in shams at 9 weeks. OPG-Fc had no effect on sPTH or Ca(2+) levels throughout the 9-week study, indicating that SHPT was a renal effect independent of bone changes. At 3 weeks, 5/6 Nx-veh rats had osteopenia compared with sham-veh rats and 5/6 Nx-OPG-Fc rats had significantly higher percent changes in whole-body BMC, leg BMD, and lumbar BMD versus 5/6 Nx-veh rats. By 6-9 weeks, elevated sPTH was associated with reversal of bone loss and osteitis fibrosa in the proximal tibial metaphysis. OPG-Fc decreased this sPTH-driven high bone turnover, resulting in augmented thickness of proximal tibial trabeculae in 5/6 Nx rats. Thus, RANKL inhibition with OPG-Fc can block the deleterious effects of continuously elevated sPTH on bone, suggesting that RANKL may be an important therapeutic target for protecting bone in patients with CKD and SHPT. PMID- 16362460 TI - Rapid establishment of chemical and mechanical properties during lamellar bone formation. AB - The development of prophylaxes and treatments of bone diseases that can effectively increase the strength of bone as a structure necessitates a better understanding of the time course by which chemical properties define the stiffness of the material during primary and secondary mineralization. It was hypothesized that these processes would be relatively slow in the actively growing skeleton. Seven-week-old Sprague-Dawley female rats (n = 8) were injected with multiple fluorochrome labels over a time span of 3 weeks and killed. Chemical and mechanical properties of the tibial mid-diaphysis were spatially characterized between the endocortical and periosteal surface by in situ infrared microspectroscopy and nanoindentation. The phosphate-to-protein ratio of bone 2-6 days old was 20% smaller at the periosteal surface and 22% smaller at the endocortical surface (P < 0.05 each) compared to older intracortical regions. The ratios of carbonate to protein, crystallinity, type A/type B carbonate, collagen cross-linking, and bone elastic modulus did not differ significantly between bone 2-6, 10-14, and 8-22 days old and intracortical regions. Intracortical properties of 10-week-old rats, except for the carbonate-to-protein ratio which was 23% smaller (P < 0.01), were not significantly different from intracortical matrix properties of young adult rats (5 months, n = 4). Spatially, the phosphate-to protein ratio (R(2) = 0.33) and the phosphate-to-carbonate ratio (R(2) = 0.55) were significantly correlated with bone material stiffness, while the combination of all chemical parameters raised the R(2) value to 0.83. These data indicate that lamellar bone has the ability to quickly establish its mechanical and chemical tissue properties during primary and secondary mineralization even when the skeleton experiences rapid growth. PMID- 16362461 TI - Predicting the risk of fracture at any site in the skeleton: are all bone mineral density measurement sites equally effective? AB - The ability to assess a patient's risk of fracture is fundamental to the clinical role of bone densitometry. Fracture discrimination is quantified by the relative risk (RR), defined as the increased risk of fracture for a 1 standard deviation decrease in bone mineral density (BMD). The larger the value of RR, the more effective measurements are at identifying patients at risk of fracture. Epidemiological studies show that RR values for predicting the risk of any fracture are approximately the same for all BMD measurement sites. In this study, we show theoretically that this interesting observation is predictable and a consequence of two related observations: (1) that fracture prediction by BMD measurement sites distant from the fracture site is quantitatively explained by the correlation of BMD measurements and (2) that all correlation coefficients between distant BMD sites are comparable, with values in the range r = 0.55-0.65. The first of these conditions (referred to as the correlation hypothesis) is important because it sets a lower limit on the RR values at distant BMD sites on the assumption that measurements at these sites contain no independent information about fracture risk over and above that provided by their correlation with the fracture site BMD. If the correlation hypothesis is true, the present study points to the importance of the correlation coefficient between BMD sites as a key index that is indicative of the ability of different types of measurement to predict fracture risk. If, on the contrary, the correlation hypothesis is not valid, there is scope to improve bone densitometry by further studies to better identify those measurements that do provide independent information about fracture risk and how best to integrate this information with existing techniques to improve decision making. PMID- 16362463 TI - Vitamin D: a necessity for children and adolescents in Greece. AB - Children and adolescents with the high bone turnover comprise a high risk population for vitamin D insufficiency. A sample of 178 clinically healthy children aged 3 to 18 years who came from public schools and lived in North West of Greece participated in the study. They were grouped into three age groups (I: 3-10, II: 11-14 and III: 15-18 years of age). Blood samples were taken during winter and summer months for determining calciotropic hormones, calcium, phosphate and biochemical markers of bone synthesis.A high percentage (47%) of the subjects aged 15-18 years was found to have 25OHD <10 ng/ml in winter but much less (13-14%) of the younger ages (13-14 years), while in the summer they were all >10 ng/ml. The prevalence was even higher in the girls of the older group accompanied by lower Pi concentrations again in winter (win:1.19+/-0.03, sum:1.93+/-0.03 mmol/l, p < 0.001). The 24,25(OH)(2)D levels were changing in parallel to 25OHD, but again in the older subjects, during winter, they were by 2/3 lower than the summer ones (0.73+/-0.10 vs. 2.41+/-0.20 ng/ml, p < 0.001). No significant differences were found between seasons and groups in the 1,25(OH)(2)D levels. The biochemical markers of bone synthesis, osteocalcin (OC) and total alkaline phosphatase (ALP), were found significantly lower in the girls of the older group both in winter and summer respectively. Even in a sunny country like Greece the adolescents living in an urban area are in high risk for vitamin D deficiency during winter. Supplementation with vitamin D of milk, of popular beverages and perhaps some foods would be of help. PMID- 16362462 TI - Geranylgeranylacetone inhibits formation and function of human osteoclasts and prevents bone loss in tail-suspended rats and ovariectomized rats. AB - Vitamin K is used for protecting against osteoporosis. Recently, it has been reported that the inhibitory effect of vitamin K(2) (menatetrenone) on bone resorption may be related to its side chain. Geranylgeranylacetone (GGA), known as teprenone, an antiulcer drug, has almost the same chemical structure as that of the side chain of menatetrenone. We hypothesized that GGA also has an inhibitory effect on osteoclastogenesis both in vitro and in vivo. GGA in pharmacological concentrations directly inhibited osteoclastogenesis from human monocytes induced by soluble receptor activator of nuclear factor-kappaB ligand. In addition, GGA induced degradation of actin rings in mature osteoclasts, which was reversed by adding geranylgeranylpyrophosphatase. Moreover, GGA increased the bone mineral density of total femur, proximal metaphysis, and diaphysis of femur in ovariectomized rats. GGA also prevented bone loss induced by hindlimb unloading in tail-suspended rats. These results indicate that GGA prevents bone loss by maintaining a positive balance of bone turnover through suppression of both the formation and the activity of osteoclasts. Thus, GGA could be used to prevent and improve osteoporosis. PMID- 16362464 TI - Excision of internal jugular vein for catheter-related thrombophlebitis. AB - This report describes a case of internal jugular vein (IJV) thrombophlebitis refractory to intravenous (IV) antibiotics and anticoagulation managed with ligation and excision of the vein. Our patient was immunosuppressed for a transplanted organ and developed a catheter-related infection following major pelvic surgery. His symptoms worsened, and failure of medical management prompted operative intervention. Excision of the inflamed, thrombosed vein led to complete resolution of his symptoms. PMID- 16362465 TI - Brain abscess associated with isolated left superior vena cava draining into the left atrium in the absence of coronary sinus and atrial septal defect. AB - A previously healthy 12-year-old girl presented with severe headache for 2 weeks. On physical examination, there was finger clubbing without apparent cyanosis. Neurological examination revealed only papiledema without focal neurologic signs. Cerebral magnetic resonance imaging showed the characteristic features of brain abscess in the left frontal lobe. Cardiologic workup to exclude a right-to-left shunt showed an abnormality of the systemic venous drainage: presence of isolated left superior vena cava draining into the left atrium in the absence of coronary sinus and atrial septal defect. This anomaly is rare, because only a few other cases have been reported. PMID- 16362466 TI - Simulation devices in interventional radiology: caveat emptor. PMID- 16362467 TI - Quality assurance guidelines for percutaneous vertebroplasty. PMID- 16362468 TI - A new method of selective, rapid cooling of the brain: an experimental study. AB - PURPOSE: To determine whether retrograde perfusion of cooled blood into one internal jugular vein (IJV) in the pig can selectively reduce the brain temperature without affecting the core body temperature (CBT). METHODS: In 7 domestic pigs, the left IJV was catheterized on one side and a catheter placed with the tip immediately below the rete mirabile. Thermistors were placed in both brain hemispheres and the brain temperature continuously registered. Thermistors placed in the rectum registered the CBT. From a catheter in the right femoral vein blood was aspirated with the aid of a roller pump, passed through a cooling device, and infused into the catheter in the left IJV at an initial rate of 200 ml/min. RESULTS: Immediately after the start of the infusion of cooled blood (13.8 degrees C) into the IJV, the right brain temperature started to drop from its initial 37.9 degrees C and reached 32 degrees C within 5 min. By increasing the temperature of the perfusate a further drop in the brain temperature was avoided and the brain temperature could be kept around 32 degrees C during the experiment. In 4 of the animals a heating blanket was sufficient to compensate for the slight drop in CBT during the cooling period. CONCLUSIONS: We conclude that brain temperature can be reduced in the pig by retrograde perfusion of the internal jugular vein with cooled blood and that the core body temperature can be maintained with the aid of a heating blanket. PMID- 16362469 TI - Experience with minimally invasive esophagectomy. AB - BACKGROUND: Minimally invasive esophagectomy (MIE) is an evolving surgical alternative to traditional open esophagectomy. Despite considerable technical challenges, it was hypothesized that MIE could be performed effectively by surgeons experienced in open esophageal resection and advanced laparoscopic surgery. The authors report their experience with 25 patients who underwent MIE for esophageal disease. METHODS: A multidisciplinary esophageal cancer team evaluated all the patients enrolled in this institutional review board-approved retrospective review study. Over an 18-month period, 25 consecutive patients (22 men and 3 women; mean age, 62 years; range, 48-77 years) with resectable esophageal cancer underwent MIE. Six patients were treated with neoadjuvant chemoradiotherapy. The preoperative diagnoses were adenocarcinoma (64%, n = 16), high-grade dysplasia (20%, n = 5), and squamous cell cancer (16%, n = 4). The outcomes evaluated included operative course, hospital and intensive care unit lengths of stay, pathologic stage, morbidity, and mortality. RESULTS: Two patients required conversion to open esophagectomy. Operative mortality was 4% (n = 1). The mean operative time was 350 min (range, 300-480), and the average blood loss was 200 ml. The patients remained ventilated for a median of 12 h, and the median intensive care unit utilization was 1 day. The median hospital length of stay was 9 days (range, 6-33 days). Major complications occurred in 32% of the patients. The anastomotic leak rate was 12%. Minor pulmonary complications occurred in 32% and atrial fibrillation in 16% of the patients. An anastomotic stricture developed in 24% of all the patients. One patient showed a positive proximal margin in the final pathology results. CONCLUSIONS: Minimally invasive esophagectomy is a technically challenging procedure that can be performed safely at the Virginia Piper Cancer Institute. Optimal results require appropriate patient selection and a multidisciplinary team experienced in the management of esophageal cancer. PMID- 16362470 TI - Histologic and clinical outcome after laparoscopic Nissen fundoplication for gastroesophageal reflux disease and Barrett's esophagus. AB - BACKGROUND: The effectiveness of laparoscopic Nissen fundoplication for the regression of Barrett's esophagus in gastroesophageal reflux disease remains controversial. The aim of this study, therefore, was to review endoscopic findings and clinical changes after laparoscopic Nissen fundoplication for gastroesophageal reflux disease, particularly for patients with Barrett's esophagus. METHODS: From September 1995 through June 2004, 127 patients with gastroesophageal reflux disease underwent laparoscopic Nissen fundoplication. All the patients had clinical and endoscopic follow-up evaluation. We further analyzed the course of 37 consecutive patients with Barrett's esophagus (29% of all laparoscopic fundoplications performed in our institution) using endoscopic surveillance with appropriate biopsies and histologic evaluation. The median follow-up period for all the patients after fundoplication was 34 months (range, 3-108 months). The median follow-up period for the patients with Barrett's esophagus was 19 months (range, 3-76 months). RESULTS: During the 9-year period, 70 women (55 %) and 57 (45%) men were treated with laparoscopic Nissen fundoplication. The median age of these patients was 42 years (range, 7-81 years). The clinical results were considered excellent for 67 patients (53%), good for 51 patients (40%), fair for 7 patients (6%), and poor for 2 patients (1%). Endoscopic surveillance showed regression of the macroscopic columnar segment in 23 patients with Barrett's esophagus (62%). Regression at a histopathologic level occurred for 15 patients (40%). The histopathology remained unchanged for 14 patients with Barrett's esophagus (38%). CONCLUSION: Laparoscopic Nissen fundoplication effectively controls intestinal metaplasia and clinical symptoms in the majority of patients with Barrett's esophagus. PMID- 16362471 TI - Endocrine surgical technique: endoscopic thyroidectomy via the lateral approach. AB - BACKGROUND: Minimal access approaches are increasingly used in endocrine surgery. Several minimal access approaches to the thyroid gland have been described, including a small-incision lateral approach and a video-assisted central approach, but to date no technique has been universally accepted. METHODS: Benefiting from the experience of more than 500 endoscopic parathyroidectomies via a lateral neck approach, the authors developed an endoscopic thyroidectomy based on the same approach and principles. Patients with solitary nodules smaller than 3 cm in diameter and no history of neck surgery or irradiation were offered this operation. A detailed description of the surgical technique is provided. RESULTS: Of the 742 thyroidectomies performed in 2004, 38 (5.1%) were endoscopic thyroidectomies. The mean nodule size was 22-mm (range, 7-47-mm), and the mean operating time was 99 min (range, 64-150-min). In all cases, the recurrent laryngeal nerve was preserved intact, and the superior and inferior parathyroids were identified, respectively, in 36 and 33 of the 38 patients. Two patients required conversion to an open cervicotomy. All patients were discharged the day after surgery. CONCLUSIONS: The described endoscopic lateral approach combines the coherence of the minimal access lateral approach and the benefits of fiberoptic magnification. It is a safe and effective technique in the hands of an appropriately trained surgeon. PMID- 16362472 TI - SLiC technique. A novel approach to percutaneous gastrostomy. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) has been an invaluable method for obtaining long-term enteral access and represents one of the first forays in the field of minimally invasive surgery. However, the traditional "pull" method for PEG tube placement continues to have some disadvantages, especially in patients with near-obstructive head and neck cancers. METHODS: We describe a new "SLiC" technique for establishing percutaneous gastrostomy using a radially expandable trocar. RESULTS: This technique is initially developed and refined on a porcine model and then successfully implemented on five human patients. CONCLUSION: The SLiC technique can be done safely and efficiently with a pediatric-sized gastroscope and avoids the need for radiation from fluoroscopy. It is a good alternative for obtaining enteral access in patients who would otherwise not be well suited for a traditional PEG tube. PMID- 16362473 TI - Immediate preoperative laparoscopic staging for squamous cell carcinoma of the esophagus. AB - BACKGROUND: Conventional preoperative staging for esophageal carcinoma could be inaccurate. Laparoscopy has been applied for the staging of various upper gastrointestinal malignancies. It can identify peritoneal and liver deposits not shown by imaging, and could reduce the number of nontherapeutic laparotomies. This study aimed to evaluate the efficacy of laparoscopic staging for the management of squamous cell carcinoma involving the mid and distal esophagus. METHODS: A retrospective review was performed for all patients with esophageal cancer evaluated for surgical resection from January 1998 to January 2004. Laparoscopy was performed for all the patients with mid and distal esophageal cancer immediately before open gastric mobilization. The efficacy of laparoscopy for the management of squamous cell carcinoma of the esophagus was evaluated. RESULTS: Among the 63 patients with potentially resectable disease shown on conventional imaging, 54 (84%) underwent esophagectomy with curative intent after laparoscopic staging. Seven patients (11%) underwent laparoscopy alone because of abdominal metastases (n = 5) or other medical conditions (n = 2) that precluded esophagectomy. Two patients (3%) had exploratory right thoracotomy without esophagectomy despite normal laparoscopic findings. The sensitivity and specificity of laparoscopic staging were 100% in this series of patients (100% sensitivity and specificity means no false-positives or -negatives). CONCLUSION: Laparoscopic staging is valuable for the management of patients with mid and distal squamous cell carcinoma of the esophagus. Patients with metastatic disease and those with prohibitive surgical risk can thus avoid unnecessary laparotomy and be offered other treatment methods. PMID- 16362474 TI - Vascular pedicle ligation techniques during laparoscopic colectomy. A prospective randomized trial. AB - BACKGROUND: A variety of devices are available for pedicle ligation during laparoscopic colectomy including vascular staplers, clips, and electrothermal bipolar vessel-sealing devices. This study assesses their speed, reliability, and cost to guide surgeons in their choice for intracorporeal pedicle ligation. METHODS: A prospective randomized study comparing laparoscopic vascular staplers and disposable clip appliers (S/C) with the LigaSure Atlas (LIG) was performed during elective right, left, and total colectomy. Cases were stratified by procedure. Failure was defined as any bleeding after proper pedicle ligation. RESULTS: The study included 48 S/C patients and 52 LIG patients with no differences in demographics, diagnosis, procedure, number of vessels ligated per procedure, or operative time. Failure occurred for 14 (9.2%) of the 152 vessels ligated in the S/C group, as compared with 5 (3%) of the 169 vessels ligated in the LIG group (p = 0.02). The median blood loss associated with device failure was 50 ml (range, 20-50 ml) in S/C group, as compared with 100 ml (range 25-800 ml) in the LIG group (p = 0.054). Major blood loss attributable to device failure and surgeon error occurred in only one LIG case. The mean cost per case of vessel ligation was significantly less in the LIG group (317 dollars +/- 0 dollars vs 400 dollars +/- 112 dollars; p < 0.001). The cost differences were greatest for total colectomy (LIG = 317 dollars +/- 0 dollars vs S/C = 565 dollars +/- 67 dollars; p = 0.002). CONCLUSION: Device failure, although more common in the S/C group, does not result in significant blood loss. The LigaSure Atlas is more cost effective during laparoscopic colectomy, especially total colectomy, and may allow the surgeon more versatility in its application. PMID- 16362475 TI - Does aggressive hydration reverse the effects of pneumoperitoneum on renal perfusion? AB - BACKGROUND: Although pneumoperitoneum (PP) decreases renal blood flow, it remains unclear whether this impacts renal function. Our aim was to characterize the effects of PP on renal perfusion and function using two fluid strategies for intravenous fluid administration. METHODS: Twelve 30-kg pigs were randomized into two groups: maintenance (3 cc/kg/h of normal saline (NS)) and bolus (15 cc/kg/h + 20 cc/kg NaCl bolus prior to induction of PP). Pigs were studied in a blinded fashion for 30 min prior, 60 min during, and 30 min after release of 15 mmHg CO(2) PP. Cardiac output (CO) and stroke volume (SV) were measured using an esophageal Doppler probe, renal cortical perfusion (RCP) was measured with a laser Doppler probe on the right kidney, and renal function was measured using the fractional excretion of sodium (FeNa) and urine output. Statistical analysis was performed with area-under-the-curve (AUC) analysis and analysis of variance RESULTS: AUC analysis revealed moderate effect size for CO (0.416) and small effect size for SV (0.366) and RCP (0.363), with decreases seen in the control group but not the bolus group. During PP, urine output increased in the bolus group (p = 0.04) but not in the control group; there was no difference in FeNa in either group. CONCLUSION: Aggressive fluid hydration during CO(2) PP of 15 mmHg preserves CO, SV, and RCP while increasing urine output. No effect on renal function as measured by FeNa was observed in either group. PMID- 16362476 TI - National analysis of in-hospital resource utilization in choledocholithiasis management using propensity scores. AB - BACKGROUND: Two treatment options exist for choledocholithiasis (CDL): endoscopic retrograde cholangiopancreatography (ERCP) and common bile duct exploration (CBDE). Resource utilization measured by total in-hospital charges (THC) and length of stay (LOS) was compared using the propensity score (PS). In this study, PS was the probability that a patient received CBDE based on comorbidities and demographics. The power of this method lies in balancing groups on variables by PS, resulting in 90% bias reduction and improved inferential validity compared to traditional analytic techniques. METHODS: Laparoscopic cholecystectomy (LC) patients with CDL who had ERCP or CBDE were identified in the 2002 U.S. Nationwide Inpatient Sample. Patients were ordered into five PS balanced strata. Mean THC, LOS, and estimated costs were compared. A linear regression model was used to estimate the contribution that LOS had on estimated costs. Monetary values were adjusted to 2004 dollars. RESULTS: A total of 40,982 patients underwent LC with CDL in 2002; 27,739 had either ERCP (93%) or CBDE (7%). Mean age was 52.7 +/- 0.4 years, with 74% women. Mean THC were less for CBDE (25,200 dollars +/- 1,800 dollars) than for ERCP (29,900 dollars +/- 800 dollars, p < 0.05). Mean LOS was less for CBDE (4.9 +/- 0.2 days) than for ERCP (5.6 +/- 0.1 days, p < 0.05). PS adjusted analysis revealed an estimated overall cost savings of 4,500 dollars +/- 1,600 dollars and reduced LOS (0.6 +/- 0.2 days) per hospitalization for CBDE. Mean THC, LOS, and estimated costs across PS score balanced strata were generally higher in the ERCP group compared to the CBDE group. LOS contributed 53% to increased THC and 62% of estimated costs. A higher cumulative incidence of complications was evident with CBDE (0.5-4.6%) compared to ERCP (0.3-3.6%). CONCLUSIONS: Based on this PS analysis, CBDE incurs less THC, reduces LOS, and has less estimated costs for CDL compared to ERCP. Furthermore, CBDE appears to be dramatically underutilized. PMID- 16362477 TI - A versatile dual-channel carbon dioxide (CO2) insufflator for various CO2)applications. The prototype. AB - BACKGROUND: Carbon dioxide (CO2), with its rapid absorptive nature, has been proven superior to atmospheric air as an insufflating agent in various clinical settings. However, CO2 insufflation has not gained wide clinical acceptance, mainly because there has been no suitable feeding system. The authors therefore have developed a versatile "dual-channel" CO2 insufflator that facilitates wider use of CO2. The objectives of this study were to introduce the authors' prototype insufflator, to evaluate its safety and performance, and to validate CO2 application using the prototype. METHODS: The prototype insufflator provides one CO2 inlet connected to a regular CO2 gas cylinder and two CO2 outlets positioned on the front and back of the device, respectively. The CO2 gas fed from the cylinder is pressure-regulated and divided into two independent conduits inside the device. The front outlet feeds CO2 gas for pneumoperitoneum at an electronically controlled pressure and flow rate. The back channel supplies CO2 gas at a fixed flow rate, allowing manual control of insufflation for various purposes. The device was evaluated with canine models. RESULTS: The prototype was safe and performed well. The CO2 application (colonoscopy in this series) using the back channel was feasible while intact CO2 pneumoperitoneum was simultaneously maintained via the front channel. There were no device malfunctions. The serial abdominal x-rays indicated that intraluminal CO2 insufflation such as that used for CO2 colonoscopy caused less residual intestinal gas than conventional air insufflation. CONCLUSIONS: The dual-channel CO2 insufflator enabled two different modes of CO2 insufflation at the same time from a single CO2 cylinder. The authors are now improving the prototype to allow safer and wider usage of CO2 in the operating room. PMID- 16362478 TI - Laparoscopic radiofrequency ablation of hepatic cavernous hemangioma. A preliminary experience with 27 patients. AB - BACKGROUND: Radiofrequency ablation (RFA), currently used extensively for liver tumors, also has been applied successfully to hepatic cavernous hemangioma (HCH) percutaneously. The aim of this study was to assess the feasibility, safety, and efficacy of laparoscopic RFA for patients with HCHs. METHODS: Between March 2001 and March 2004, 27 patients with symptomatic and rapid-growth lesions were treated by laparoscopic RFA using the RF-2000 generator system. The treatment related complications were observed. All the patients were followed up with helical computed tomography scans and ultrasonography at regular intervals to assess the therapeutic efficacy of laparoscopic RFA. RESULTS: This study assessed 9 men and 18 women with a mean age of 41.6 +/- 8.3 years. Three additional intrahepatic lesions missed preoperatively were found in three patients on intraoperative ultrasound. A total of 27 patients with 50 liver lesions were treated successfully with laparoscopic RFA. The mean maximum tumor diameter was 5.5 +/- 2.0 cm. The mean length of time for RFA per lesion was 20.7 +/- 11.9 min, and the mean blood loss was 134.4 +/- 88.9 ml. Laparoscopic cholecystectomy was performed simultaneously for gallstones in 13 patients and for abutting of gallbladder from hemangioma in 2 patients. In addition, 3 patients also had a laparoscopic deroofing of simple hepatic cysts. Although postoperative low-grade fever and transient elevation of serum transaminase levels were observed in 13 patients, there were no complications related to laparoscopic RFA. During a median follow-up period of 21 months (range, 12-42 months), complete lesion necrosis was achieved for all the patients. CONCLUSIONS: Laparoscopic RFA therapy is a safe, feasible, and effective treatment option for patients with symptomatic and rapid-growth HCHs located on the surface of the liver or adjacent to the gallbladder. Intraoperative ultrasonography is a useful adjunct for detecting additional liver lesions and offering more accurate targeting for RFA. PMID- 16362479 TI - Prospective randomized trial of laparoscopic gastrojejunostomy versus duodenal stenting for malignant gastric outflow obstruction. AB - BACKGROUND: We prospectively compared laparoscopic gastrojejunostomy with duodenal stenting as a means of palliating malignant gastric outflow obstruction. METHODS: A total of 27 patients with malignant gastric outflow obstruction were randomized to either laparoscopic gastrojejunostomy (LGJ) or duodenal stenting (DS) over a 3-year period. RESULTS: Thirteen patients underwent successful LGJ and 10 had successful DS. Eight patients had complications after LGJ, but none had complications after DS. Patients who underwent LGJ had a significant increase in visual analog pain score at day 1 (p = 0.05), and also had a longer hospital stay compared to those who underwent DS (11.4 vs. 5.2 days, p = 0.02). After DS, patients experienced an improvement in physical health at 1 month as measured using the Short Form-36 (SF-36) questionnaire (p < 0.01). There was no change following LGJ. CONCLUSION: Duodenal stenting is a safe means of palliating malignant gastric outflow obstruction. It offers significant advantages for patients compared with minimal-access surgery. PMID- 16362480 TI - Does laparoscopy worsen the prognosis for incidental gallbladder cancer? AB - BACKGROUND: More than 75% of cholecystectomies are done laparoscopically and less than one-third of gallbladder carcinomas are known presurgically. It is supposed that the laparoscopic technique could adversely affect the prognosis of gallbladder cancer. METHODS: The C-A-E has started a register of all cases of cholecystectomy with a postoperative incidental finding of gallbladder carcinoma. The aim is to compare the prospectively collected follow-up data on the outcome of these patients and to answer the question of whether laparoscopic cholecystectomy affects the prognosis of incidental gallbladder cancer. RESULTS: A total of 377 cases have been recorded so far. These include 201 patients treated by the laparoscopic procedure, 119 by an open procedure, and 57 by an intraoperative conversion. The survival shows a significantly better life expectancy for the patients treated laparoscopically. CONCLUSION: The life expectancy is higher for the laparoscopically treated patients and this cannot be explained by the fact that the laparoscopic technique is used to treat the earlier stages of cancer. The access technique does not seem to influence the prognosis for gallbladder carcinomas. PMID- 16362481 TI - Electrical stimulation for gastroparesis. Gastric motility restored. AB - BACKGROUND: Gastroparesis is a disabling, and sometimes fatal, disease that often does not respond to medical treatment. This single-surgeon prospective study examines the safety and 6-month efficacy of electrical stimulation for the treatment of gastroparesis. METHODS: Sixteen patients with medically refractory gastroparesis underwent laparoscopic implantation of an electrical stimulator device (Enterra Therapy, Medtronic, Minneapolis, MN, USA) consisting of a subcutaneous stimulator and two gastric wall leads. Gastric emptying scans (GES) confirmed the diagnosis of gastroparesis. Patients were evaluated preoperatively using a self-administered GI symptomatology questionnaire and RAND 36 Health Survey. Once patients were >6-months from implantation, a repeat GES was obtained and patients completed a postoperative GI symptomatology questionnaire and RAND 36 Health Survey. Ten of 16 patients in this case series were >6-months from implantation. One was lost to follow-up. An F-test was used to establish equality of standard deviations between the 16 patients evaluated preoperatively and the subset of 10 patients evaluated postoperatively. A Student's t-test was used to evaluate the significance of differences in pre- and postoperative results. RESULTS: Average operating time was 117 min with no intraoperative complications. The majority of patients were discharged on postoperative day 1. There were two complications in the postoperative period. Patients experienced a significant decrease in nausea and vomiting as measured by the GI symptomatology questionnaire. Half of all patients no longer required gastric prokinetic medications and there was a subjective reduction of pyrosis, early satiety, and epigastric pain. A significant increase in quality of life as measured by the RAND 36 Health Survey was seen, and six of eight patients no longer demonstrated gastroparesis on GES. CONCLUSION: Laparoscopic implantation of an electrical stimulation device is a safe and effective treatment by subjective and objective standards for the management of medically refractory gastroparesis. PMID- 16362482 TI - Skin wound closure with a novel shape-memory alloy fixator. AB - BACKGROUND: Previously, we have described novel, thermally deployable tissue fixators based on the shape-memory properties of nickel titanium. The present study reports on the in vivo wound closure with these fixators as a preliminary to evaluating their use for bowel anastomoses. METHODS: Twenty adult Sprague Dawley rats had 2.0-cm dorsal skin incisions approximated either with shape memory alloy (SMA) fixators or conventional skin staples on a random basis. Electrical resistance heating was used to transform and deploy the SMA fixators into the wound. The rats were killed on day 14 or day 28 and wound specimens were harvested for force distraction studies and histologic examination. RESULTS: There was no incidence of wound dehiscence. 14- and 28-day wounds from both groups showed no significant difference in breaking force or energy. Histology revealed appropriate stages of wound healing for both SMA-closed and control wounds. CONCLUSIONS: The results confirm the efficacy and safety of tissue-edge approximation with SMA fixators. PMID- 16362483 TI - A novel method distinguishes between mutation rates and fixation biases in patterns of single-nucleotide substitution. AB - Analysis of the genome-wide patterns of single-nucleotide substitution reveals that the human GC content structure is out of equilibrium. The substitutions are decreasing the overall GC content (GC), at the same time making its range narrower. Investigation of single-nucleotide polymorphisms (SNPs) revealed that presently the decrease in GC content is due to a uniform mutational preference for A:T pairs, while its projected range is due to a variability in the fixation preference for G:C pairs. However, it is important to determine whether lessons learned about evolutionary processes operating at the present time (that is reflected in the SNP data) can be extended back into the evolutionary past. We describe here a new approach to this problem that utilizes the juxtaposition of forward and reverse substitution rates to determine the relative importance of variability in mutation rates and fixation probabilities in shaping long-term substitutional patterns. We use this approach to demonstrate that the forces shaping GC content structure over the recent past (since the appearance of the SNPs) extend all the way back to the mammalian radiation approximately 90 million years ago. In addition, we find a small but significant effect that has not been detected in the SNP data-relatively high rates of C:G-->A:T germline mutation in low-GC regions of the genome. PMID- 16362484 TI - Molecular evolution and phylogeny of sipunculan hemerythrins. AB - We sequenced seven new hemerythrin (Hr) and myohemerythrin (myoHr) cDNAs from Sipunculus nudus and Golfingia vulgaris vulgaris, thus providing new comparative data that significantly increase the set of the known Hr and myoHr sequences. Bayesian inference, maximum likelihood, and maximum parsimony phylogenetic analyses were performed to investigate the evolutionary relationships among the sipunculan and annelid Hr and myoHr sequences. Annelid myoHrs and sipunculan Hrs were resolved as monophyletic groups. Conversely sipunculan myoHrs did not form a clade. The Hrs having an octameric quaternary structure were resolved as a monophyletic group. The octameric cluster includes the Hr sequences of G. v. vulgaris, Themiste zostericola, Themiste discriptum, and Phascolopsis gouldii. Siphonosoma cumanense Hr, which has a trimeric quaternary structure, assumes a sister group position of the octameric clade. The S. nudus Hrs, having a quaternary structure that is not well resolved, assume an isolate position within the Hrs clade. Likelihood-based analyses reveal that purifying selection mainly characterized the evolution of Hr and myoHr. We suggest that starting from a common gene ancestor, two distinct quaternary structures evolved in the sipunculan Hrs and this differentiation was probably favored by the acquisition of distinct physiological advantages. PMID- 16362485 TI - Incompatible land uses and the topology of cumulative risk. AB - The extensive literature on environmental justice has, by now, well defined the essential ingredients of cumulative risk, namely, incompatible land uses and vulnerability. Most problematic is the case when risk is produced by a large aggregation of small sources of air toxics. In this article, we test these notions in an area of Southern California, Southeast Los Angeles (SELA), which has come to be known as Asthmatown. Developing a rapid risk mapping protocol, we scan the neighborhood for small potential sources of air toxics and find, literally, hundreds of small point sources within a 2-mile radius, interspersed with residences. We also map the estimated cancer risks and noncancer hazard indices across the landscape. We find that, indeed, such large aggregations of even small, nondominant sources of air toxics can produce markedly elevated levels of risk. In this study, the risk profiles show additional cancer risks of up to 800 in a million and noncancer hazard indices of up to 200 in SELA due to the agglomeration of small point sources. This is significant (for example, estimates of the average regional point-source-related cancer risk range from 125 to 200 in a million). Most importantly, if we were to talk about the risk contour as if they were geological structures, we would observe not only a handful of distinct peaks, but a general "mountain range" running all throughout the study area, which underscores the ubiquity of risk in SELA. Just as cumulative risk has deeply embedded itself into the fabric of the place, so, too, must intervention seek to embed strategies into the institutions and practices of SELA. This has implications for advocacy, as seen in a recently initiated participatory action research project aimed at building health research capacities into the community in keeping with an ethic of care. PMID- 16362486 TI - Incentive-based conservation programs in developing countries: a review of some key issues and suggestions for improvements. AB - Biodiversity conservation in developing countries has been a challenge because of the combination of rising human populations, rapid technological advances, severe social hardships, and extreme poverty. To address the social, economic, and ecological limitations of people-free parks and reserves, incentives have been incorporated into conservation programs in the hopes of making conservation meaningful to local people. However, such incentive-based programs have been implemented with little consideration for their ability to fulfill promises of greater protection of biodiversity. Evaluations of incentive-based conservation programs indicate that the approach continually falls short of the rhetoric. This article provides an overview of the problems associated with incentive-based conservation approaches in developing countries. It argues that existing incentive-based programs (IBPs) have yet to realize that benefits vary greatly at different "community" scales and that a holistic conceptualization of a community is essential to incorporate the complexities of a heterogeneous community when designing and implementing the IBPs. The spatial complexities involved in correctly identifying the beneficiaries in a community and the short-term focus of IBPs are two major challenges for sustaining conservation efforts. The article suggests improvements in three key areas: accurate identification of "target" beneficiaries, greater inclusion of marginal communities, and efforts to enhance community aptitudes. PMID- 16362487 TI - Developing a national tiger action plan for The Union of Myanmar. AB - A century ago, tigers were considered pests in Myanmar. Hunters claimed thousands, yet populations persisted. In the past century, because of habitat loss and prey depletion, coupled with the recent demand for traditional medicines, tiger populations have been reduced to a few hundred individuals. As a first step toward long-term planning for tigers, and to guide efforts to increase protected area coverage, the Myanmar government in 1998 initiated a project to develop a revised National Tiger Action Plan. Extensive surveys confirmed tigers in only 4 of 17 survey locations. Significant populations are thought to persist in the far North and far South of the country, where large, intact forests offer the potential for tiger recovery. With partnerships and collaborations, tiger populations can be protected in the short term (<5 years) by expanding protected areas and corridors, mobilizing enforcement staff to reduce poaching of tigers and prey, and amending existing wildlife legislation in accordance with international laws. Over the long term (5-20 years), recovery of Myanmar's tiger populations will depend on increasing support from local people, zoning tiger areas to reduce habitat loss and disturbance, and maintaining connectivity of existing national and transboundary forests. This article reviews the development of a new National Tiger Action Plan for the Union of Myanmar and discusses a blueprint for conservation measures aimed at saving tigers from extinction. PMID- 16362488 TI - The impotence of cumulative effects assessment in Canada: ailments and ideas for redeployment. AB - Cumulative effects assessment (CEA) in Canada is in dire straits. Despite a huge amount of talk and a flurry of developmental activity associated with CEA concepts, it has not lived up to its glowing promise of helping to achieve sustainability of diverse valued ecosystem components. This article aims to articulate that failure, to examine it in terms of six major problems with CEA, and to propose solutions. The six problem areas include (1) application of CEA in project-level environmental impact assessments (EIAs), (2) an EIA focus on project approval instead of environmental sustainability, (3) a general lack of understanding of ecologic impact thresholds, (4) separation of cumulative effects from project-specific impacts, (5) weak interpretations of cumulative effects by practitioners and analysts, and (6) inappropriate handling of potential future developments. We advocate improvements not only within the purview of project specific EIAs, but also mainly in the domain of region-scale CEAs and regional environmental effects frameworks (or perhaps land use planning). Only then will the CEA begin to approach the promise of securing sustainability of valued ecosystem components. PMID- 16362489 TI - Effectiveness of a coral-derived surfacing material for reducing sediment production on unpaved roads, Schoffield barracks, Oahu, Hawaii. AB - This study evaluated the effectiveness of two application rates of a coral derived surfacing material for both traffic and nontraffic road conditions using simulated rainfall (110-120 mm h(-1) for 30-90 min) on 0.75-m (wide) x 5.0-m (long) plots of similar slope (roughly 0.1 m m(-1)). The coral is a locally available material that has been applied to unpaved roads surfaces on Schoffield Barracks, Oahu, Hawaii (USA), where this experiment was conducted. The simulations show that compared with a bare control plot, the coral-based surface application rates of 80 and 160 kg m(-2) (equivalent to only 10- and 20-mm thicknesses) reduced road sediment production by 75% and 95%, respectively, for nontraffic conditions. However, after two passes of the research vehicle during wet conditions, sediment production rates for the two coral treatments were not significantly different from those on the bare road plots. The overall effectiveness of the coral-derived surfacing material is unsatisfactory, primarily because the on-road surface thickness associated with the application rates tested was too small. These rates were selected to bracket those applied to training roads in the study area. Furthermore, the composition of the coral-based material does not facilitate the development of a sealed, erosion-resistant surface. When applied at the low rates tested, the coral material breaks down under normal traffic conditions, thereby losing its ability to counter shearing forces exerted by overland flow on long hillslopes where erosion measures are most needed. These simulations, combined with observations on roads in the study area, indicate that this material is not an appropriate road surfacing material for the site-at least for the low application rates examined. These results are preliminary; extended testing of higher applications rates at the hillslope scale under natural climate and traffic conditions is needed to better judge the effectiveness of this material over time. PMID- 16362490 TI - Colorado river floods, droughts, and shrimp fishing in the upper gulf of California, Mexico. AB - Accurate procedures that measure hydrologic variability would have great value for evaluating ecosystem impacts of upstream water use in the Colorado River Basin. Many local extractive income-based stakeholders rely directly or indirectly on ecosystem health and are adversely affected when the river does not flow. This study focuses on the impact of little or no Colorado River flow on the Mexican shrimp industry. Although there have been complaints that U.S. diversions of Colorado River flow have greatly impaired the shrimp fishery, this research demonstrates that freshwater rarely reaches the Gulf even during times of flooding, and that other factors such as overfishing may influence the instability of shrimp populations. Advanced very-high-resolution radiometer (AVHRR) satellite imagery was used to assess water volumes diverted away from the channel of the Colorado River and ultimately the Gulf of California during flooding periods. Analysis of data demonstrated that little freshwater actually reaches the Gulf even during floods because of its diversion into a large dry lake bed basin known as Laguna Salada. Fuller use of the Colorado River throughout its entire course to the sea is possible and could benefit a large cohort of users without catastrophic habitat destruction in delta ecosystems. Reconstruction of a natural earthen berm, as proposed by Ducks Unlimited, would maximize the use of floodwaters for ecosystem benefits. These findings have profound implications for local economic activities dependent on hydrologic resources in the Colorado River Delta and Upper Gulf. PMID- 16362491 TI - Assessment of environmental water demands (EWD) of forests for two distinct Indian ecosystems. AB - Sustainable use of water and land resources requires that these scarce resources be appropriately allocated among various competing human activities. Worldwide, there is a realization now that sustainable river basin management should be accorded the highest priority, because it deals not only with technical, but also with ecological and socioeconomic aspects, and thus calls for a multidisciplinary and integrated approach. However, most of the policy and planning documents have either remained silent, or have made only implicit reference to the importance of environmental water demand (EWD) and its quantification. Therefore, in the light of its importance, a methodology has been evolved in this article for quantifying EWD for various forested areas in two distinctly different Indian river basins: Brahmani (humid zone) and Sabarmati (dry zone). The article analyzes and discusses EWD estimates at three different spatial levels: river basins, states, and districts within them, and finally presents a comparative analysis of all these results. Findings of the present study will be immensely useful in understanding various ecological issues connected with water resource projects and proposals in these river basins. PMID- 16362492 TI - Managing endangered species within the use-preservation paradox: the Florida manatee (Trichechus manatus latirostris) as a tourism attraction. AB - A significant challenge to wildlife managers in tourism settings is to provide visitors with opportunities to observe rare and endangered wildlife while simultaneously protecting the target species from deleterious impacts. Nearly 100,000 people annually visit Crystal River, Florida, USA to observe and swim with the Florida manatee, an endangered species. This research aimed to investigate and describe human-manatee interactions in a tourism context, to understand the salient issues related to such interactions as identified by stakeholders, and to recommend a course of action to address multiple interests in the planning and management of human-manatee interactions. Five issues were identified by all stakeholder groups: water quality, harassment, density and crowding, education, and enforcement. Currently, the U.S. Fish and Wildlife Service, which is responsible for manatee management, does not have mechanisms in place to manage the tourism component of the manatee encounter. Although a regulatory approach can be taken, a better approach would be to create an organization of tour operators to establish "best practices" that reflect the goal of the managing agency to enhance manatee protection (and thus ensure their livelihood) and to enhance the visitor experience. PMID- 16362493 TI - The RpoS-mediated regulation of isocitrate dehydrogenase gene expression in Escherichia coli. AB - The Escherichia coli NADP(+)-dependent isocitrate dehydrogenase (IDH; EC 1.1.1.42), encoded by an icd gene, is a tricarboxylic acid (TCA) cycle enzyme responsible for the oxidative decarboxylation of isocitrate to alpha ketoglutarate. In order to examine how the icd gene expression is regulated, an icd-lacZ reporter fusion was constructed. While the icd gene was induced in exponential growth phase, it was repressed in stationary growth phase. Genetic inactivation of an rpoS gene, whose product is an alternative sigma factor, induced the icd gene expression approximately 4.8 times more in the stationary phase and the IDH enzyme activity in the rpoS mutant was 3.2 times higher than that in the wild type, indicating that the RpoS factor acts as a negative regulator of the icd gene expression in the stationary phase. PMID- 16362494 TI - Response of wheat seedlings to ni stress: effects of supplemental calcium. AB - The effect of excess Ni (1 mM Ni) on wheat plants as well as the role of Ca (1 mM Ni+5000 microM Ca) for amelioration of toxicity and recovery of growth and photosynthesis in Ni-stressed wheat was evaluated. Growth, nutrient status (Ca, Mg, Fe, K, Na), and photosynthesis showed a distinct decrease strictly related to the period of treatment. Calcium ameliorated to a certain extent toxic symptoms of Ni, due to antagonistic action between Ni and Ca ions. Since chlorophyll content and variable fluorescence (Fv) decreased significantly, but Fo did not particularly change, the decrease of t1/2 with increasing duration of Ni exposure indicates negative changes on the acceptor side of PSII, which also may result from diminution of Calvin cycle. The maximum quantum yield for energy trapping was also suppressed. Plant transfer to Hoagland solution+5000 microM Ca caused recovery to plant morphology and physiology. Even in control plants, during recovery period an increased Ca concentration in plant tissues with concomitant increased rates of growth and morphology was observed. Ni concentration in plants exposed to 1 mM Ni+5000 microM Ca was lower than in plants exposed to 1 mM Ni. In all treatments a certain increase of plant nutrients was observed during recovery. PMID- 16362495 TI - Fate of the chemical warfare agent VX in asphalt: a novel approach for the quantitation of VX in organic surfaces. AB - VX is one of the most toxic chemical warfare agents. Its low volatility and its persistence in the environment raise the issue of long-term exposure risks, either by inhalation or by transdermal penetration. Therefore, a topic of acute interest is the fate of VX in preservative environmental surfaces. In this work, the fate of VX in asphalt pavement, a suspected preservative matrix, was explored, by applying a novel quantitative method for the extraction of trapped VX from "digested" asphalt. It is based on dissolution of asphalt in toluene, precipitation of the heavy components by basic methanol followed by GC-NPD analysis. This method is complementary to methanol extraction of VX from the outer surface of asphalt, and enabled us to explore the total amount of viable VX both on and inside the matrix. Using this method, bis-diisopropylaminoethyl disulfide [(DES)2], a degradation product of VX, was also assayed. Small chunks of Asphalt were spiked with VX, sealed and analyzed after various aging periods up to 425 days. The level of VX on the outer surface of the asphalt was found to be diminishing with time following a single-exponential decay. The level inside the asphalt increases during the first day, decays steeply to a level of about 5% during the following two weeks, and declines moderately during all the period up to 425 days following a bi-exponential decay. The total recovery of VX from the asphalt declined from almost 100% after 30 minutes to about 2% after 425 days, with a half-life of about 14 days. PMID- 16362496 TI - Effect of various copper supplements to feed of laying hens on cu content in eggs, liver, excreta, soil, and herbage. AB - Copper is often added to poultry diets as an antimicrobial agent at doses greatly exceeding the nutritional requirement. In this study, the basal diet of laying hens containing 9.2 mg Cu/kg was supplemented with CuSO(4) x 5H(2)O at 0, 25, 65, 115, and 240 mg Cu/kg dry matter (DM). At Cu dietary concentration just below the level permitted by the European Union (35 mg/kg), the Cu content in the egg yolk was significantly (p < 0.05) increased by 26%, and nonsignificantly by 4.1, 9.1, and 7.9% in the egg white, eggshell, and liver, respectively. When Cu concentration in the diet was doubled, the effect of Cu on Cu content in eggshell and liver was statistically significant as well. In no liver sample was the hygienic limit of Cu content (80 mg/kg) exceeded. Supplementation of diets with Cu increased Cu concentration in excreta linearly from 25.3 to 396.8 mg/kg DM. Dried excreta were used for fertilization of grassland at 21 g N/m(2). Three months later, soil and herbage were sampled and analyzed. The Cu concentration in soil increased from 25.3 to only 46.4 mg/kg DM when dietary Cu concentration rose from 9.2 to 243.7 mg Cu/kg DM. Corresponding Cu concentrations in herbage were 6.8 and 19.2 mg/kg DM. It can be concluded that the deposition of Cu in eggs and liver of hens fed Cu-supplemented diets does not represent a hygienic risk. The accumulation of Cu in soil fertilized with excreta of Cu-fed hens and in herbage was limited. PMID- 16362499 TI - Role of voltage-gated potassium channels in cancer. AB - Ion channels are being associated with a growing number of diseases including cancer. This overview summarizes data on voltage-gated potassium channels (VGKCs) that exhibit oncogenic properties: ether-a-go-go type 1 (Eag1). Normally, Eag1 is expressed almost exclusively in tissue of neural origin, but its ectopic expression leads to uncontrolled proliferation, while inhibition of Eag1 expression produces a concomitant reduction in proliferation. Specific monoclonal antibodies against Eag1 recognize an epitope in over 80% of human tumors of diverse origins, endowing it with diagnostic and therapeutic potential. Eag1 also possesses unique electrophysiological properties that simplify its identification. This is particularly important, as specific blockers of Eag1 currents are not available. Molecular imaging of Eag1 in live tumor models has been accomplished with dye-tagged antibodies using 3-D imaging techniques in the near-infrared spectral range. PMID- 16362500 TI - N-methyl-D-aspartate receptor in human prostate cancer. AB - Expression of the N-methyl-D-aspartate receptor (NMDAr) and its involvement in cellular proliferation is well-known in tumors of neuronal tissue, such as glioma and neuroblastoma. We have investigated NMDAr expression in the normal, hyperplastic and neoplastic human prostate by immunohistochemistry. Low stromal NMDAr immunostaining was observed in 2 of 12 (17%) normal prostate specimens, but epithelial NMDAr staining was not seen. Of 18 benign prostatic hyperplasia (BPH) specimens, none had stromal NMDAr staining, but 2 had low and 1 had high epithelial NMDAr immunoreactivity. Moderate to high NMDAr immunostaining was observed in the stroma of 60 of 145 (41%) prostate cancer (PCa) specimens. Epithelial NMDAr staining was low in 26 (18%) and moderate to high in 36 (25%) of 145 PCa specimens. We have also examined the effects of the NMDAr antagonist memantine on the growth of ten human cancer cell lines: four prostate, two breast and four colon. The NMDAr antagonist memantine inhibited in-vitro growth of all ten cell lines, with half-maximal growth-inhibition at 5 to 20 microg/ml (23 to 92 microM) memantine. An NMDA agonist, L-cysteinesulfinic acid, stimulated cellular proliferation of all ten cell lines, with maximal growth-stimulation (30% to 75%, depending on the cell line) observed between doses of 33 to 66 microM. Our data provide evidence for the expression and activity of NMDAr in prostate cancer. PMID- 16362502 TI - Roles of volume-sensitive Cl- channel in cisplatin-induced apoptosis in human epidermoid cancer cells. AB - The anti-cancer drug cisplatin induces apoptosis by damaging DNA. Since a stilbene-derivative blocker of Cl-/HCO3- exchangers and Cl- channels, SITS, is known to induce cisplatin resistance in a manner independent of intracellular pH and extracellular HCO3-, we investigated the relation between cisplatin-induced apoptosis and Cl- channel activity in human adenocarcinoma KB cells. A stilbene derivative, DIDS, reduced cisplatin-induced caspase-3 activation and cell death, which were detected over 18 h after treatment with cisplatin. DIDS was also found to reduce sensitivity of KB cells to 5-day exposure to cisplatin. Whole-cell patch-clamp recordings showed that KB cells functionally express volume-sensitive outwardly rectifying (VSOR) Cl- channels which are activated by osmotic cell swelling and sensitive to DIDS. Pretreatment of the cells with cisplatin for 12 h augmented the magnitude of VSOR Cl- current. Thus, it is concluded that cisplatin induced cytotoxicity in KB cells is associated with augmented activity of a DIDS sensitive VSOR Cl- channel and that blockade of this channel is, at least in part, responsible for cisplatin resistance induced by a stilbene derivative. PMID- 16362501 TI - Ca2+ signaling, intracellular pH and cell volume in cell proliferation. AB - Mitogens control progression through the cell cycle in non-transformed cells by complex cascades of intracellular messengers, such as Ca2+ and protons, and by cell volume changes. Intracellular Ca2+ and proton concentrations are critical for linking external stimuli to proliferation, motility, apoptosis and differentiation. This review summarizes the role in cell proliferation of calcium release from intracellular stores and the Ca2+ entry through plasma membrane Ca2+ channels. In addition, the impact of intracellular pH and cell volume on cell proliferation is discussed. PMID- 16362505 TI - Clinical relevance of ion channels for diagnosis and therapy of cancer. AB - Ion channels have a critical role in cell proliferation and it is well documented that channel blockers can inhibit the growth of cancer cells. The concept of ion channels as therapeutic targets or prognostic biomarkers attracts increasing interest, but the lack of potent and selective channel modulators has hampered a critical verification for many years. Today, the knowledge of human ion channel genes is almost complete and molecular correlates for many native currents have already been identified. This information triggered a wave of experimental results, identifying individual ion channels with relevance for specific cancer types. The current pattern of cancer-related ion channels is not arbitrary, but can be reduced to few members from each ion channel family. This review aims to provide an overview of the molecularly identified ion channels that might be relevant for the most common human cancer types. Possible applications of these candidates for a targeted cancer therapy or for clinical diagnosis are discussed. PMID- 16362506 TI - Is there a relationship between the pH and volume of saliva and esophageal pH metry results? AB - There are several implications in the buffer capacity and in the protective role of saliva in gastroesophageal reflux disease (GERD) and other digestive disorders. The lack of knowledge about the production and quality of saliva and the fact that saliva plays an important role in digestive homeostasis motivated the authors to study the relationship between reflux measured by esophageal pH metry and the pH and volume of saliva in individuals with GERD and its laryngopharyngeal manifestations (LPR). The study was designed as a randomized clinical trial. The studied population consisted of 39 adults with GERD and LPR confirmed by a positive 24-hour double-probe esophageal pH-metry. Unstimulated whole saliva was collected and its pH and volume were compared to pH-metry results. Patients were divided into four groups according to the number of distal episodes of reflux and two groups according to the presence or absence of proximal reflux. A highly significant difference was found between the groups with distal reflux when comparing salivary volume alone and salivary volume/salivary pH. The same occurred for the presence or absence of proximal reflux. These results suggest a direct correlation between salivary volume and "salivary pH x volume" with the number of distal and proximal episodes of reflux on the esophageal pH-metry. If larger studies confirm this finding, in the future it might be possible to diagnose GERD and LPR through a simple "spit test," avoiding more costly and invasive procedures. PMID- 16362503 TI - Ion channels in cell proliferation and apoptotic cell death. AB - Cell proliferation and apoptosis are paralleled by altered regulation of ion channels that play an active part in the signaling of those fundamental cellular mechanisms. Cell proliferation must--at some time point--increase cell volume and apoptosis is typically paralleled by cell shrinkage. Cell volume changes require the participation of ion transport across the cell membrane, including appropriate activity of Cl- and K+ channels. Besides regulating cytosolic Cl- activity, osmolyte flux and, thus, cell volume, most Cl- channels allow HCO3- exit and cytosolic acidification, which inhibits cell proliferation and favors apoptosis. K+ exit through K+ channels may decrease intracellular K+ concentration, which in turn favors apoptotic cell death. K+ channel activity further maintains the cell membrane potential, a critical determinant of Ca2+ entry through Ca2+ channels. Cytosolic Ca2+ may trigger mechanisms required for cell proliferation and stimulate enzymes executing apoptosis. The switch between cell proliferation and apoptosis apparently depends on the magnitude and temporal organization of Ca2+ entry and on the functional state of the cell. Due to complex interaction with other signaling pathways, a given ion channel may play a dual role in both cell proliferation and apoptosis. Thus, specific ion channel blockers may abrogate both fundamental cellular mechanisms, depending on cell type, regulatory environment and condition of the cell. Clearly, considerable further experimental effort is required to fully understand the complex interplay between ion channels, cell proliferation and apoptosis. PMID- 16362504 TI - Ion channels and cancer. AB - Membrane ion channels are essential for cell proliferation and appear to have a role in the development of cancer. This has initially been demonstrated for potassium channels and is meanwhile also suggested for other cation channels and Cl- channels. For some of these channels, like voltage-gated ether a go-go and Ca2+-dependent potassium channels as well as calcium and chloride channels, a cell cycle-dependent function has been demonstrated. Along with other membrane conductances, these channels control the membrane voltage and Ca2+ signaling in proliferating cells. Homeostatic parameters, such as the intracellular ion concentration, cytosolic pH and cell volume, are also governed by the activity of ion channels. Thus it will be an essential task for future studies to unravel cell cycle-specific effects of ion channels and non-specific homeostatic functions. When studying the role of ion channels in cancer cells, it is indispensable to choose experimental conditions that come close to the in vivo situation. Thus, environmental parameters, such as low oxygen pressure, acidosis and exposure to serum proteins, have to be taken into account. In order to achieve clinical application, more studies on the original cancer tissue are required, and improved animal models. Finally, it will be essential to generate more potent and specific inhibitors of ion channels to overcome the shortcomings of some of the current approaches. PMID- 16362507 TI - Intrinsic properties of the adult human mylohyoid muscle: neural organization, fiber-type distribution, and myosin heavy chain expression. AB - The mylohyoid (MH) muscle plays a critical role in chewing, swallowing, respiration, and phonation. The present study was designed to test the hypothesis that the functional properties of the MH are reflected by its intrinsic specializations, including the neural organization, fiber-type distribution, and myosin heavy chain (MHC) expression. Adult human MH muscles were investigated to determine the nerve supply pattern using Sihler's stain, banding pattern and types of motor endplates using acetylcholinesterase (AChE) staining and silver impregnation, and muscle fiber type and MHC composition using immunocytochemical and immunoblotting techniques. The adult human MH was found to have the following neuromuscular specializations. First, the muscle was innervated by several branches of the MH nerve derived from the mandibular division of the trigeminal nerve. Each of the nerve branches supplied a distinct region of the muscle, forming a segmental innervation pattern. Second, the MH had a single motor endplate band which was located in the middle of the muscle length. Both en plaque and en grappe types of motor endplates were identified on the MH muscle fibers. Finally, the adult human MH fibers expressed unusual MHC isoforms (i.e., slow-tonic, alpha-cardiac, embryonic, and neonatal) which coexisted with the major MHC isoforms (i.e., slow type I, fast type IIa, and fast type IIx), thus forming various major/unusual (or m/u) MHC hybrid fiber types. The m/u hybrid fibers (84% of the total fiber population) were the predominant fiber types in the adult MH muscle. Determination of the neuromuscular specializations of the MH is helpful for better understanding of the muscle functions and for development of strategies to treat MH-related upper airway disorders. PMID- 16362508 TI - An acoustic profile of normal swallowing. AB - Cervical auscultation has been proposed as a technique to augment the clinical evaluation of dysphagia to improve its accuracy in the diagnosis of dysphagia. Before using cervical auscultation to reliably diagnose disordered swallowing, it is necessary to first acoustically characterize normal swallowing for comparison with dysphagic swallowing. Ninety-seven healthy adult participants consumed teaspoon boluses of various consistencies while the sounds of swallowing were recorded. Descriptive statistics were reported for measures of duration, intensity, and frequency of the acoustic swallowing signal. Correlations between the variables and between bolus consistencies were computed. Overall, results compared favorably with previous research. Significant correlations were found among several of the variables, including an increasing duration of the acoustic swallowing signal with increasing age and decreasing intensity of the signal with increasing age. None of the variables differed significantly as a function of gender. Of potential clinical relevance, significant correlations between bolus consistencies for the duration and intensity variables indicated relative similarities across bolus consistencies. Duration and intensity of the acoustic signal appeared to be the most reliable of the variables measured. These results could serve as a reference point for future studies into normal swallowing across multiple bolus consistencies and volumes and eventually be compared with disordered swallowing. PMID- 16362509 TI - Effects of a sour bolus on the intramuscular electromyographic (EMG) activity of muscles in the submental region. AB - A sour bolus has been used as a modality in the treatment of oropharyngeal dysphagia based on the hypothesis that this stimulus provides an effective preswallow sensory input that lowers the threshold required to trigger a pharyngeal swallow. The result is a more immediate swallow onset time. Additionally, the sour bolus may invigorate the oral muscles resulting in stronger contractions during the swallow. The purpose of this investigation was to compare the intramuscular electromyographic activity of the mylohyoid, geniohyoid, and anterior belly of the digastric muscles during sour and water boluses with regard to duration, strength, and timing of muscle activation. Muscle duration, swallow onset time, and pattern of muscle activation did not differ for the two bolus types. Muscle activation time was more tightly approximated across the onsets of the three muscles when a sour bolus was used. A sour bolus also resulted in a stronger muscle contraction as evidenced by greater electromyographic activity. These data support the use of a sour bolus as part of a treatment paradigm. PMID- 16362511 TI - The relationship between observations and measures of oral and pharyngeal residue from videofluorography and scintigraphy. AB - We examined measures of oral and pharyngeal residues from scintigraphic studies and estimates/observations from videofluorographic (modified barium swallow) studies taken on the same day but not concurrently in 16 dysphagic patients of varying etiologies presenting with oral and/or pharyngeal dysphagia. Oral and pharyngeal residuals following the swallow were quantified scintigraphically and were then compared with measures of residuals obtained from the modified barium swallow. Estimates of oral and pharyngeal residues from the modified barium swallows were generated by a trained observer who was blinded to the scintigraphic data. Positive and significant Spearman correlations between oral and pharyngeal residue measures from scintigraphy and observations of oral and pharyngeal residues from modified barium swallows were found. This supports the validity of observations of oral and pharyngeal residues in clinical studies. Limitations of these observations are discussed. PMID- 16362510 TI - Silent aspiration: what do we know? AB - Although clinically evident aspiration is common in subjects with dysphagia, a significant proportion may aspirate silently, i.e., without any outward signs of swallowing difficulty. This article reviews the literature on the prevalence, etiology, and prognostic significance of silent aspiration. An electronic database search was performed using silent aspiration, aspiration, dysphagia, and stroke as search terms, together with hand-searching of articles. Silent aspiration has been described in many conditions and subgroups of patients (including normal individuals), using a number of detection methods, making comparisons a challenge. The best data are for acute stroke, in which 2%-25% of patients may aspirate silently. Mechanisms associated with silent aspiration may include central or local weakness/incoordination of the pharyngeal musculature, reduced laryngopharyngeal sensation, impaired ability to produce a reflexive cough, and low substance P or dopamine levels. In terms of prognosis, silent aspiration has been associated with increased morbidity and mortality in many but not all studies. However, some degree of silent aspiration at night may be normal in healthy individuals. The phenomenon of silent aspiration is poorly understood and further research is needed to improve methods of detection and thereby better define its prevalence and prognostic significance. PMID- 16362512 TI - Odynophagia secondary to variant thyroid cartilage anatomy. AB - We present a case of a displaced superior cornu of the thyroid cartilage causing odynophagia and a foreign body sensation of the pharynx. Diagnosis was made by computed tomography (CT) scan and confirmed by direct endoscopic examination. We review the literature that addresses possible etiologies of this variant anatomy and review treatment options. PMID- 16362513 TI - Laryngeal phonation after tracheoesophageal diversion. AB - When tracheoesophageal diversion is performed, laryngeal phonation is usually impossible. We, however, experienced the case of a 65 year-old female patient, who was able to do so. Fluoroscopic and videoendoscopic examinations elucidated her mechanism of phonation. This is the first detailed report describing the mechanism of laryngeal phonation after the procedure. Our case is compared with a few other similar cases found in available literature. PMID- 16362514 TI - Swallow-induced syncope in a patient with achalasia. AB - A 48-year-old man was referred for the evaluation of dysphagia and syncope. He suffered from both dysphagia and transient loss of consciousness when eating solid food, especially bread. An upper gastrointestinal barium examination and esophageal manometry suggested achalasia. Ambulatory ECG revealed marked sinus bradycardia when experiencing a syncopal episode following the ingestion of a solid meal. A permanent pacemaker was implanted and the patient's syncopal attacks were relieved following this procedure. However, the dysphagia and cardiac arrhythmia completely disappeared only following a successful pneumatic balloon dilatation for achalasia. This observation suggests that swallowing induced sinus bradycardia and syncope might occur in the course of achalasia and these rhythm abnormalities could be relieved by successful treatment of the achalasia. PMID- 16362516 TI - A morphometric analysis of the Japanese macaque (Macaca fuscata) mandibular cheek teeth from the Torihama Shell-midden, Early Jomon Period, Fukui Prefecture, Japan. AB - We quantitatively examined the differences in the size and proportion of the Japanese macaque (Macaca fuscata) by comparing the Early Jomon specimens from Torihama shell-midden, Fukui Prefecture and modern specimens from Fukui Prefecture. The purpose of this study was to explore the temporal change in the proportion and size of teeth of the Japanese macaques based on the quantified data. The result of measurements of lower premolars and molars demonstrated that sexual dimorphism was evident only among the modern specimens where the females were significantly smaller than males. The size of male Torihama specimens was within the range of the modern population, whereas the size of the female Torihama specimens was significantly larger than the modern female population. The proportional pattern of premolars and molars for male and female Torihama specimens also differed. The results may suggest a possible difference in the degree of size reduction between males and females since the last glacial period. PMID- 16362517 TI - The rpoH gene encoding heat shock sigma factor sigma32 of psychrophilic bacterium Colwellia maris. AB - The rpoH gene encoding a heat shock sigma factor, sigma(32), was cloned from the psychrophilic bacterium Colwellia maris. The deduced amino acid sequence of sigma(32) from C. maris is more than 60% homologous to that of sigma(32) from mesophilic bacteria. The RpoH box, a 9-amino-acid sequence region (QRKLFFNLR) specific to sigma(32), and two downstream box sequences complementary to a part of 16S rRNA were identified. Primer extension analysis showed that the C. maris rpoH is expressed from only one sigma(70)-type promoter. Northern blot analysis showed that the level of rpoH mRNA was clearly increased at 20 degrees C, a temperature that induces heat shock in this organism. In the presence of an inhibitor of transcriptional initiation, the degradation of rpoH mRNA was much slower at 20 degrees C than at 10 degrees C. Thus, increased stability of the rpoH mRNA might be responsible for the rpoH mRNA accumulation. The predicted secondary structure of the 5'-region of C. maris rpoH mRNA was different from the conserved patterns reported for most mesophilic bacteria, and the base pairing of the downstream boxes appeared to be less stable than that of Escherichia coli rpoH mRNA. Thus, essential features that ensure the HSP expression at a relatively low temperature are embedded in the rpoH gene of psychrophiles. PMID- 16362518 TI - Percutaneous extracorporeal cardiopulmonary support: current practice and its role. PMID- 16362519 TI - Update on mitral valve surgery. AB - Recently, the outcome for patients with mitral valve disease has significantly improved. This may be due to concomitant advances in many fields. In particular, the development of surgical techniques has contributed to this improvement, and many surgical techniques and topics are introduced in this article. After the evaluation of results in mitral valve surgery and exploration of the relationship between the mitral valve and the mitral subvalvular apparatus, it is currently accepted that mitral valve repair is superior to replacement and that replacement with preservation of the mitral subvalvular apparatus is preferable to replacement alone. Another current topic is mitral repair in dilated cardiomyopathy. Mitral regurgitation is a known complication of end-stage cardiomyopathy and is associated with a poor prognosis because of progressive mitral annular dilatation. Recently, to solve this problem, undersized mitral annuloplasty or cardiomyoplasty has been advocated. In the future, mitral valve surgery may be performed off-pump or by a percutaneous approach. Several groups are investigating the use of new devices and new techniques that avoid cardiopulmonary bypass in experimental studies. In this article, we review current topics and discuss our experiences in mitral valve surgery. PMID- 16362520 TI - Hemolysis caused by surface roughness under shear flow. AB - In this study, the relationship between the degree of roughness of blood contact surfaces under laminar shear flow conditions and the level of hemolysis resulting from this roughness was investigated using a rotational shear stressor. Unlike previous in vitro experiments that used a pumped circuit, the level of hemolysis was directly evaluated under a constant shear flow. In total, 1.8% of the blood contact area was roughened to an arithmetic mean roughness (Ra) value of between 0.4 and 9.2 microm by machine processing and a shear load was applied for 30 min at a shear flow rate of 3750 s(-1). As a result, the threshold Ra value for the induction of hemolysis was found to be between 0.4 and 0.8 microm. In addition, the results of this experiment suggested that the high shear stress resulting from surface roughness plays a major role in determining the level of hemolysis caused by surface roughness. PMID- 16362521 TI - Segmented polyurethane modified by photopolymerization and cross-linking with 2 methacryloyloxyethyl phosphorylcholine polymer for blood-contacting surfaces of ventricular assist devices. AB - To improve the biocompatibility of pulsatile ventricular assist devices (VADs), the blood-contacting surface of the segmented polyurethane (SPU) diaphragm employed in an electromechanical VAD was modified by introducing 2 methacryloyloxyethyl phosphorylcholine (MPC) units into its surface and forming an interpenetrating polymer network (IPN) structure, which contained independently cross-linked MPC polymer and SPU. The SPU diaphragm modified with an IPN structure was then assembled into a target test pump and underwent continuous pump operation at 37 degrees C for 2 weeks in a simulated systemic circulation using a mock circulatory loop. The surface characteristics of the pump diaphragm after 2 weeks of pump operation were then analyzed with an X-ray photoelectron spectroscope (XPS) and gold-colloid-labeled immunoassay. The XPS surface analysis of the IPN-modified SPU indicated the firm anchoring of MPC units even after 2 weeks of pump operation (the phosphor : carbon ratio was reduced by only 0.09%). The IPN-modified diaphragm prevented protein adsorption as well as cell adhesion in comparison to the unmodified SPU surface. This result thus validated that (1) the IPN structure could firmly secure MPC units to the SPU surface even in a high-mechanical-stress and high-shear environment, (2) the antithrombogenic power of MPC units remained unchanged after 2 weeks of continuous exposure to a high-shear environment, and (3) the IPN modified SPU cross-linked with MPC could be a powerful antithrombogenic surface for blood pumps used for chronic circulatory support of cardiac patients. PMID- 16362522 TI - Preparation and properties of ProNectin F-coated biodegradable hollow fibers. AB - ProNectin F-coated biodegradable hollow fibers were newly prepared and their cytocompatibility was evaluated in vitro. Although the coating efficiency onto poly(L-lactic acid) (PLLA) and poly(lactide-co-caprolactone) [p(LA/CL)] matrices was similar, the cell adhesion properties were greatly affected by the nature of the polymer substrate. ProNectin F-coated PLLA showed about seven times higher cytocompatibility than ProNectin F-coated p(LA/CL). The single-extruded melt spinning method and the core-sheath bicomponent melt spinning method were employed to prepare PLLA hollow fibers. The effect of the spinning conditions, such as the melt draw ratio, spinneret temperature, and take-up speed, on the diameter and wall thickness of the spun fibers was studied in detail. For single extruded melt spinning, a segmented type of spinneret was used, and the effect of the flow rate of nitrogen, which was confined in the hollow part of fibers, was studied. X-ray photographs of the drawn hollow fibers, clarified the significant molecular orientation, which was much higher than that in drawn solid PLLA fiber under identical drawing conditions. The morphology and mechanical properties of hollow fibers demonstrated an increase in the tensile strength and a decrease in the thickness of the PLLA wall with increased nitrogen flow rates and melt draw ratios for single-extruded melt spinning. These results indicate the unique characteristics of ProNectin F-coated PLLA hollow fibers, which can be successfully utilized as a biodegradable substrate. PMID- 16362523 TI - Evaluation of endocrine disrupting activity of plasticizers in polyvinyl chloride tubes by estrogen receptor alpha binding assay. AB - Polyvinyl chloride (PVC) tubing is an indispensable medical material for extracorporeal circulation therapy. However, di(2-ethylhexyl)phthalate (DEHP), a suspected endocrine disruptor, can be eluted from PVC, suggesting that an alternative material that does not contain DEHP is needed for clinical applications. First, we evaluated the endocrine disrupting risks of the plasticizers contained in PVC tubes by investigating their binding affinities for the human estrogen receptor alpha (ERalpha). Our results revealed that, while DEHP has some binding affinity for ERalpha, neither epoxidized soybean oil nor tris(2-ethylhexyl)trimellitate (an alternative to DEHP) has any affinity for ERalpha. Second, we evaluated the endocrine disrupting risks of a tube made of newly developed plasticizer-free (PF) materials. We confirmed the presence of DEHP and detected several unidentified substances in plasma stored within the PVC tube. This plasma's competitive binding affinity for ERalpha was significantly higher than that of control plasma (P < 0.01). In contrast, the profile of plasma stored in the PF tube was similar to that of the control, both in terms of high performance liquid chromatography chromatograms and competitive binding capacity for ERalpha, suggesting that the PF tube is biocompatible and is useful for reducing the elution of substances capable of binding to ERalpha. PMID- 16362524 TI - Hollow-fiber blood-dialysis membranes: superoxide generation, permeation, and dismutation measured by chemiluminescence. AB - The interaction of blood with a material surface results in activation of the body's humoral immune system and the generation of reactive oxygen species (ROS). It has recently become clear that ROS are central to the pathology of many diseases. In this study, we evaluated the superoxide generation, permeation, and dismutation in hollow-fiber dialysis membranes by using 2-methyl-6-p methoxyphenylethynyl-imidazopyrazinone (MPEC) as a superoxide-reactive chemiluminescence producer and an optical fiber probe to detect the resulting chemiluminescence in the hollow fiber lumen. We measured the superoxide generated when bovine blood leukocytes were brought into contact with dialysis membranes. Superoxide permeation was determined by measuring MPEC chemiluminescence in the hollow fiber lumen using an optical fiber probe. Additionally, superoxide dismutation was evaluated by examining the difference in superoxide permeability for membranes with and without vitamin E coating. Superoxide generation varies for different membrane materials, depending on the membrane's biocompatibility. Superoxide permeability depends on the diffusive permeability of membranes. No marked decrease in superoxide permeability was observed among membrane materials. The superoxide permeability of vitamin E-coated membrane was smaller than that of uncoated membrane. The antioxidant property of vitamin E-coated membranes is hence effective in causing superoxide dismutation. PMID- 16362525 TI - Hemodilution with liposome-encapsulated low-oxygen-affinity hemoglobin does not attenuate hypothermic cerebral ischemia in rats. AB - Hypothermia decreases cerebral metabolism and increases hemoglobin oxygen affinity. A hypothesis that the reversal of increased oxygen affinity would further attenuate hypothermic cerebral ischemia was tested by evaluating the effects of liposome-encapsulated hemoglobin (LipoHb) with low oxygen affinity (P50 = 40-50 mmHg) on hypothermic incomplete cerebral ischemia. Wistar rats were randomly assigned to one of the following two groups: (A) exchange transfusion with LipoHb solution (Hb = 6 g/dl) (LipoHb, n = 5), (B) no exchange transfusion (control, n = 5). After surface cooling to 22 degrees C, forebrain ischemia was induced for 15 min by bilateral carotid artery occlusion combined with a decrease in the mean arterial pressure (MAP) to 40 mmHg. (31)P-magnetic resonance spectroscopy was performed during ischemia and 45 min of reperfusion. After reperfusion, MAP was significantly higher in the control group than in the LipoHb group (P < 0.01), although there were no significant differences during ischemia. Intracellular pH and phosphocreatine (PCr) levels decreased during ischemia and returned to the preischemic level in both groups following reperfusion. The LipoHb group had a significantly larger decrease and smaller recovery in PCr than the control group (P < 0.0001). Althouth beta-adenosine triphosphate decreased during ischemia in the LipoHb group, it increased in the control group (P < 0.0001). Inorganic phosphate (Pi) increased during ischemia and decreased to the normal value after reperfusion. The LipoHb group experienced a significantly larger production of Pi than the control group (P = 0.02). Hemodilution with high P50 LipoHb does not reduce ischemic energy depletion induced by hypothermic incomplete forebrain ischemia in rats. PMID- 16362526 TI - Dysfunction of an ATS valve in the aortic position: the first reported case caused by pannus formation. AB - A 64-year-old woman underwent aortic valve replacement with a 21-mm Advancing The Standard (ATS) open-pivot mechanical heart valve for bicuspid aortic valve stenosis. In addition to the appearance of a new cardiac murmur, echocardiography performed 3 years after surgery showed a high pressure gradient across the ATS valve and a reduction in the valve orifice area. Cineradiography of the valve revealed restricted leaflet opening. Subsequent multidetector-row computed tomography clearly demonstrated pannus overgrowth on the inflow aspect of the ATS valve. During a repeat operation, subvalvular overgrown pannus was confirmed and the ATS valve was replaced with a bioprosthetic valve. This is the first reported case of prosthetic valve dysfunction resulting from pannus formation in a patient with an ATS valve in the aortic position. PMID- 16362527 TI - An association study between the transthyretin (TTR) gene and mental retardation. AB - It is known that in the pathogenesis of mental retardation (MR), both genetic and environmental factors (particularly iodine deficiency) appear to play a critical role. Transthyretin (TTR) transports between 20% and 30% of serum thyroxine in normal individuals and it is the main T(4)-binding protein in CSF. Variability in the TTR gene may influence risk for iodine-deficiency-based MR. The SNPs we selected from dbSNP were detected and identified using ARMS-PCR and sequencing methods, and we identified five novel sequence variants. Singular-locus association analysis indicated no association between the TTR gene and MR. In haplotype analysis, however, we found a haplotype CGTG+ (rs723744/G+6649C/T+6690C/rs2276382/del9) showed a weak positive association with MR (chi(2) = 6.699, p = 0.035). Finally, we concluded that the weak positive result is more likely to be due to sampling error and the small size of this haplotype resulting from its relative low frequency. Our negative results provide no evidence that variants of TTR gene influence susceptibility to MR in the iodine-deficient areas of China and suggest that there may be a compensatory mechanism(s) in humans and mice, which work(s) to compensate the effect of mutation in the TTR gene on MR. PMID- 16362528 TI - Auto-immune reactive polyserositis in meningococcal meningoencephalitis: a case report. PMID- 16362529 TI - Fluid-attenuated inversion recovery (FLAIR) sequences for the assessment of acute stroke: inter observer and inter technique reproducibility. AB - BACKGROUND AND PURPOSE: Diffusion-weighted magnetic resonance (MR) imaging (DWI), and three-dimensional (3D) time-of-flight (TOF) MR angiography (MRA), are highly sensitive for the early detection of stroke and arterial occlusion. However, only a few studies have evaluated the sensitivity of conventional MR sequences that are usually included in the imaging protocol. The aim of this study was to evaluate interobserver and intertechnique reproducibility of Fluid-Attenuated Inversion Recovery (FLAIR) sequences for the diagnosis of early brain ischemia and arterial occlusion. METHODS: Over a 30-month period, brain MR examinations were performed in 34 patients within 12 hours after stroke onset. Imaging protocol included FLAIR sequences, DWI and 3D TOF MRA. Ten observers including radiologists and neurologists, performed separately a visual interpretation of FLAIR images for the detection of brain ischemia and arterial occlusion seen as an arterial high signal. DWI and 3D TOF MRA were used as reference and interpreted independently by two senior radiologists. Interobserver agreement was assessed for image quality, detectability and conspicuity of lesions whereas intertechnique agreement was only judged for lesion detectability. RESULTS: On FLAIR sequences, interobserver agreement for the detection of brain ischemia and arterial occlusion was excellent (kappa = 0.81 and 0.87 respectively). The concordance between FLAIR and DWI sequences for the detection of brain ischemia and between FLAIR and 3D TOF MRA for the detection of arterial occlusion were judged as excellent for all observers (kappa = 0.91 and 0.89 respectively). CONCLUSION: Although DWI is the most sensitive technique with which to detect acute stroke, FLAIR imaging may also be useful to demonstrate both acute ischemia and arterial occlusion with an excellent interobserver reproducibility. PMID- 16362530 TI - No correlation between amount of aberrant transcript and severity of phenotype in hereditary spastic paraplegia patients with a c.1242A > G splice mutation in the SPG4 gene. PMID- 16362531 TI - Corpus callosum functioning in patients with normal pressure hydrocephalus before and after surgery. AB - OBJECTIVES: Our aim was to evaluate corpus callosum functioning in a group of patients with normal pressure hydrocephalus (NPH) before and after shunting. METHODS: Left ear-extinction under a dichotic listening task was evaluated in twenty-three patients with NPH, 30 patients with Alzheimer's disease and 30 aged controls. RESULTS: Patients with NPH had higher levels of left ear extinction than the control and Alzheimer's groups. Sixty-one percent of NPH patients exhibited left ear suppression, compared with 13% of Alzheimer's patients and 17% of controls. Following surgery, NPH patients showed a significant change in the degree of asymmetry in the dichotic listening task. CONCLUSIONS: Hydrocephalus was associated with left-ear extinction,which diminished after surgery. Our results may indicate reversible functional damage in the corpus callosum. PMID- 16362532 TI - Injection with methylprednisolone in patients with the carpal tunnel syndrome: a randomised double blind trial testing three different doses. AB - OBJECTIVE: To determine the efficacy of 20, 40 and 60 mg methylprednisolone injections in patients with the carpal tunnel syndrome. METHODS: Included were patients with signs and symptoms of carpal tunnel syndrome of more than 3 months duration confirmed by electrophysiological tests. Patients were in a double blind trial randomised to treatment consisting of injections proximal to the carpal tunnel with 20, 40 or 60 mg methylprednisolone. Primary outcome was improvement of symptoms requiring no further treatment. These patients were followed for one year. RESULTS: There were no significant differences in the treatment response between the three randomised groups at one-year follow-up (log rank analysis 1.51, 2 df, 0.4711). In the 20, 40 and 60 mg treatment groups, 56%, 53% and 73% of the patients respectively were free of important symptoms at six months follow up. Of the patients treated with one or two injections 22% were finally referred to surgery within one year of the first treatment. No side effects were recorded. CONCLUSION: A single local injection of methylprednisolone 20, 40 or 60 mg results in long lasting improvement in approximately half of the patients. There is a trend in favour of the highest dose. A second injection may further reduce the number of patients requiring surgery. PMID- 16362533 TI - Treatment of phobic postural vertigo. A controlled study of cognitive-behavioral therapy and self-controlled desensitization. AB - In balance clinic practice, phobic postural vertigo is a term used to define a population with dizziness and avoidance behavior often as a consequence of a vestibular disorder. It has been described as the most common form of dizziness in middle aged patients in dizziness units. Anxiety disorders are common among patients with vestibular disorders. Cognitive-behavioral therapy is an effective treatment for anxiety disorders, and vestibular rehabilitation exercises are effective for vestibular disorders. This study compared the effect of additional cognitive-behavioral therapy for a population with phobic postural vertigo with the effect of self-administered vestibular rehabilitation exercises. 39 patients were recruited from a population referred for otoneurological investigation. Treatment effects were evaluated with the Dizziness Handicap Inventory, Vertigo Symptom Scale, Vertigo Handicap Questionnaire, and Hospital Anxiety and Depression Scale. All patients had a self treatment intervention based on education about the condition and recommendation of self exposure by vestibular rehabilitation exercises. Every second patient included was offered additional cognitive behavioral therapy. Fifteen patients with self treatment and 16 patients with cognitive- behavioral treatment completed the study. There was significantly larger effect in the group who received cognitive behavioral therapy than in the self treatment group in Vertigo Handicap Questionnaire and the Hospital Anxiety and Depression scale and its subscales. Cognitive-behavioral therapy has an additional effect as treatment for a population with phobic postural vertigo. A multidisciplinary approach including medical treatment, cognitive-behavioral therapy and physiotherapy is suggested. PMID- 16362534 TI - 1,25-Dihydroxyvitamin D and 25-hydroxyvitamin D--mediated regulation of TRPV6 (a putative epithelial calcium channel) mRNA expression in Caco-2 cells. AB - BACKGROUND: TRPV6 is a member of the vanilloid subfamily of transient receptor potential (TRP) proteins and likely functions as an epithelial calcium channel in calcium-transporting organs, such as the intestine, kidney, and placenta. TRPV6 mRNA expression is strongly regulated by 1,25-dihydroxyvitamin D (1,25VD), the active hormonal form of vitamin D, in intestine and in Caco-2 cells, a human colon cancer cell line. AIM OF THE STUDY: The aim of the present study was to characterise the mode of regulation of the 1,25VD-mediated TRPV6 mRNA expression and to test the effect of the precursor of 1,25VD namely 25 hydroxyvitamin D (25 VD) on TRPV6 mRNA expression in Caco-2 cells. METHODS: Caco-2 cells were treated in a 2 x 2 format with 1,25VD and the transcriptional inhibitor actinomycin D (AD, 4 microg/ml), and also with translational inhibitor cycloheximide (CHX, 10 microg/ml) after 14 days in culture and TRPV6 mRNA levels were determined using reverse transcription-real time PCR.TRPV6 mRNA half life studies were performed by inhibiting transcription followed by sampling at various time points for TRPV6 mRNA. Varying concentrations of 25 VD were used to test their effect on TRPV6 mRNA in the presence of 5% FBS and also in the absence of serum (but containing insulin-transferrin-selenium mixture) for 24 h. RESULTS: Treatment with 10(-7) M 1,25VD for 8 h resulted in a 60-fold increase in TRPV6 mRNA and this increase could be completely blocked with AD. Treatment with CHX to inhibit de novo protein synthesis did not prevent the initiation of 1,25VD-induced TRPV6 expression, although it did reduce the extent of TRPV6 mRNA accumulation. We found that TRPV6 mRNA half-life was 8 h in Caco-2 cells and was not altered by 1,25VD treatment. Finally, we observed that treatment with 10(-6) M of the pro hormone 25 VD for 24 h resulted in a significant increase in TRPV6 expression in Caco-2 cells, which is consistent with the presence of 1alpha-hydroxylase (CYP27B1) expression in Caco-2 cells and a possible autocrine vitamin D signaling pathway in colon cells. CONCLUSIONS: 1,25 dihydroxyvitamin D regulates TRPV6 expression by a process that requires new mRNA and protein synthesis and the point of regulation lies likely at the transcriptional level especially since vitamin D did not increase the half life of TRPV6 mRNA. In addition, the prohormone form of 1,25 dihydroxyvitamin D, i. e. the 25 hydroxyvitamin D, induced TRPV6 mRNA expression in Caco-2 cells. PMID- 16362536 TI - Research highlights from the literature. PMID- 16362537 TI - Electrogastrography. PMID- 16362538 TI - Impaired circadian rhythm of gastric myoelectrical activity in patients with multiple system atrophy. AB - In order to evaluate gastric motility and its circadian rhythm in patients with multiple system atrophy (MSA) and healthy control subjects, we measured gastric myoelectrical activity (GMA) for 24 hours using a cutaneous electrogastrogram (EGG) recorder in 14 MSA patients and 9 age-matched controls. We analyzed six 10 minute segments of EGG before and after each meal and two 20-minute EGG segments during sleep; three parameters were used for the analysis: dominant frequency (DF), instability coefficient of dominant frequency (ICDF), and dominant power (DP). DF increased during daytime and decreased during sleep in the control, while this circadian variation was blunted in the patients with MSA. The average DF of the eight segments in the MSA patients did not differ from that of the control. Both MSA patients and control subjects did not show the circadian variation of ICDF and DP. The average ICDF of the eight segments in the patients with MSA was significantly decreased when compared with that of the control (p < 0.01). No differences were observed in DP between the two groups. This study indicates that the healthy subjects appear to have a circadian rhythm of DF, and the patients with MSA appear to have impaired circadian rhythm of DF and decreased ICDF possibly due to the degeneration of the central autonomic neurons. PMID- 16362539 TI - Interaction between central-peripheral chemoreflexes and cerebro-cardiovascular control. AB - We investigated the interaction between hypoxia and hypercapnia on ventilation and on cerebro-cardio-vascular control. A group of 12 healthy subjects performed rebreathing tests to determine the ventilatory response to hypoxia, at different levels of carbon dioxide (CO(2)), and to normoxic hypercapnia. Oxygen saturation (SaO(2)), end-tidal CO(2) (et-CO(2)), minute ventilation, blood pressure, R-R interval and mid-cerebral artery flow velocity (MCFV) were continuously recorded. The hypoxic ventilatory response significantly increased under hypercapnia and decreased under hypocapnia (slopes L/min/% Sa O(2): -0.33 +/- 0.05, -0.74 +/- 0.02 and -1.59 +/- 0.3, p < 0.0001, in hypocapnia, normocapnia and hypercapnia, respectively). At similar degrees of ventilation, MCFV increased more markedly during normocapnic hypoxia than normoxic hypercapnia; the slopes linking MCFV to hypoxia remained unchanged at increasing levels of et-CO(2), whereas the regression lines were shifted upward. The R-R interval decreased more markedly during normocapnic hypoxia than normoxic hypercapnia and the arterial baroreflex sensitivity was decreased only by hypoxia. Cardiovascular responses to hypoxia were not affected by different levels of et-CO(2). We conclude that concomitant hypoxia and hypercapnia, while increasing ventilation synergistically, exert an additive effect on cerebral blood flow. Increased sympathetic activity (and reduced baroreflex sensitivity) is one of the mechanisms by which hypoxia stimulates cardiac sympathetic activity. PMID- 16362540 TI - P wave duration and dispersion in multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) has been reported to be associated with extensive autonomic dysfunction as well as to involve the cardiovascular system. In this study we compared P wave duration and dispersion (PWD) of MS patients to healthy control subjects. METHOD: Thirty-one MS patients (mean age 35 +/- 9 years) and 33 healthy control subjects (mean age 34+/-7 years) were included in the study. Twelve-lead surface ECG recording (paper speed of 50 mm/s) was obtained from all participants. The change in maximum and minimum P wave duration was measured manually and the difference between the two values was defined as PWD. RESULTS: There was no difference between the two groups in terms of baseline demographic characteristics. Maximum P wave duration was higher in MS patients than controls (126 +/- 6 vs. 122 +/- 5 ms, p = 0.004). Minimum P wave duration was found to be similar in MS patients and healthy controls (72 +/- 8 vs. 75 +/- 8 ms, p = 0.2). Mean PWD value of MS patients was also found to be higher than those of controls (53 +/- 10 vs. 47 +/- 10 ms, p = 0.01). CONCLUSION: P wave duration and PWD was found to be statistically different in patients with MS and control subjects. PMID- 16362541 TI - Reproducibility of heart rate variability and blood pressure variability in individuals with spinal cord injury. AB - Individuals with spinal cord injury (SCI) are prone to orthostatic intolerance and an increased risk of cardiovascular disease. The use of heart rate variability (HRV) and blood pressure variability (BPV) as indices of cardiovascular regulation would be valuable in this population; however, their reproducibility has yet to be tested in those with SCI. The purpose of this study was to examine the day-to-day reproducibility of resting HRV and BPV in individuals with SCI. Ten individuals (age 35.9 +/- 13.2 yrs) with chronic (5.4 +/- 7.7 years post injury) SCI (C4-T12; ASIA A-C) participated. On two occasions within a two-week period, 10-minute supine electrocardiogram and Finapres blood pressure recordings were obtained during spontaneous breathing. Computer software calculated frequency domain measures of HRV and BPV (Low frequency (LF) power, High frequency (HF) power, and LF:HF ratio). Intraclass correlations coefficients (R) were used as an index of day-to-day reproducibility, and analyses were conducted on all participants and only those with tetraplegia. For HRV, measures of heart rate, LF, and LF:HF were found to be highly reproducible (R = 0.82 0.88); however, the reproducibility of HF was found to be poor (all participants: R = 0.53, tetraplegia: R = 0.66). Measures of blood pressure as well as systolic BPV also showed high reproducibility (R = 0.72-0.93). Measures of diastolic BPV were less reproducible but still acceptable (R = 0.71-0.89) with the exception of LF:HF(DBP) (R = 0.51). In conclusion, despite the autonomic dysfunction associated with SCI, measures of HRV and BPV may still be used as reproducible indices of autonomic cardiovascular regulation in this population. PMID- 16362542 TI - Stability over time of short-term heart rate variability. AB - Heart rate variability (HRV) is a widely used method to assess cardiac autonomic control. However, the reproducibility of especially short-term HRV has not been properly evaluated. Therefore, we assessed the stability of short-term HRV over a three to four month period. We had seven consecutive electrocardiographic (ECG) recordings from 89 subjects with stable coronary artery disease obtained during a large multicenter study. The HRV assessments were performed from these 40-minute ECG-recordings simulating normal daily activities, i. e., recordings consisting of 5 to 10 minute periods of rest, paced breathing, standing, submaximal exercise and recovery. Both time and frequency domain HRV analyses were conducted from the whole 40-minute recordings and from the 5-minute periods of rest and paced breathing. The coefficient of variation (CV) varied between 5.1-16.7% for the 40 minute and 6.0-37.1% for the 5-minute time domain and 4.4-11.0 % for the 40 minute and 7.2-16.5 % for the 5-minute frequency domain measurements. The mean of the RR intervals and the total power showed the highest stability over time. The most unstable measure was the standard deviation of all NN intervals (SDNN). In conclusion, most short-term HRV measures were highly stable over time indicating low physiological variation. However, the SDNN showed large variability in consecutive recordings. PMID- 16362543 TI - Autonomic reactivity and clinical severity in children with sickle cell disease. AB - Individual differences in autonomic nervous system reactivity have been studied in relation to physical and mental health outcomes, but rarely among children with chronic disease. The purpose of this study was to examine the associations among autonomic reactivity, clinical severity, family stressors, and mental health symptoms in children with homozygous sickle cell disease. Nineteen children with homozygous sickle cell disease participated in a cross-sectional study involving parent-completed measures, medical record reviews and laboratory based measures of autonomic nervous system responses to social, cognitive, physical and emotional challenges. Autonomic reactivity was significantly associated with both clinical severity and externalizing behavior symptoms. Children with greater parasympathetic withdrawal during challenges compared to rest had significantly more severe disease (r = -0.45, p < 0.05); greater sympathetic activation during challenges compared to rest was associated with more externalizing behavior symptoms ( r= 0.44, p < 0.05). Children experiencing major family stressors had internalizing behavior symptoms but no difference in autonomic reactivity or clinical severity compared to children experiencing fewer family stressors. Individual differences in autonomic reactivity may offer a new, biologically plausible account for observed variation in painful episodes, other physical complications and behavioral symptoms among children with sickle cell disease. PMID- 16362544 TI - Mental strain in medical students during simulator training measured by forehead sweating. AB - Forehead sweating developed in 78% of medical students when they attempted tracheal intubation on a simulator, and increased during subsequent attempts if the first attempt was not successful. Thus, forehead sweating may be a useful index of mental strain. PMID- 16362546 TI - QSART in idiopathic pure sudomotor failure. AB - QSART (quantitative sudomotor axon reflex testing) was performed in a patient with idiopathic pure sudomotor failure. Generalized reduction in thermoregulatory sweating and complete absence of axon reflex sweating were observed, suggesting a deficit of sweat gland cholinergic synaptic transmission or receptors. QSART responded promptly to treatment. Putative pathophysiological mechanisms are discussed. PMID- 16362545 TI - The effect of hormone replacement therapy cessation on heart rate variability in postmenopausal women. AB - Heart rate variability showed no statistically significant changes in the time, frequency or complexity measurements either before or following cessation of hormone replacement therapy in healthy postmenopausal women. PMID- 16362547 TI - Oculosympathetic hyperactivity in idiopathic hyperhidrosis. AB - We describe a patient suffering from idiopathic hyperhidrosis, more pronounced on the right side, who presented with intermittent oculosympathetic hyperactivity (mydriasis, lid retraction and more rapid pupillary dilatation) on the same side. Mydriasis was induced by stress, loss of sleep and cold pressor test. The clinical features in this patient suggest an involvement of the hypothalamic structures. PMID- 16362548 TI - Abnormal baroreflex responses in multiple sclerosis. PMID- 16362550 TI - Pyridostigmine in the treatment of orthostatic hypotension. PMID- 16362554 TI - [Age and migration--an overview]. AB - In the course of demographic change, the number of elderly migrants increases, and the population of elderly people in Germany becomes more varied-facts that have to be taken into account more intensely when it comes to planning services for elderly people in the future. The need for action is intensified by the fact that elderly migrants are still worse off than natives of the same age in many areas of life (e. g. income, health, etc.). At the same time, elderly migrants cannot automatically be regarded as helpless or isolated because of their solid family help networks.Meanwhile, on political and social levels, approaches have been developed to open services and institutions for the special needs of minority ethnic elders. Furthermore, offers have been established by the creation of ethnic-specific emphases that focus on the life style and the everyday life of the target group. In spite of all these efforts, the whole range of care services for the population of elderly migrants is still regarded as insufficient by experts. Therefore, especially in the fields of networking, cooperation and controlling of the people involved, information transfer, and training measures that aim at better preparing the provider of care services for elderly people are highly recommended in order to meet the increasing needs of help and care of older migrants more precisely. PMID- 16362555 TI - [Potentials of elderly migrants]. AB - Re-migration and immigration are the causes for the continual fluctuation within the migrant population. It is, however, currently foreseeable that the elderly migrant population living in Germany will steadily increase. Because of the large degree of heterogeneity among the elderly migrant population in Germany, the following report will be limited to a discussion of those groups of migrant workers, who originate from former recruiting countries. These groups will constitute the majority of elderly migrants in the coming years. Among the first generation of migrants, some groups have extensive ethnic social networks. This is mostly the case with respect to the migrants from Turkey. These have access to their own ethnic infrastructure, and, due to this fact, there is little need for the initiation of social contacts outside their own ethnic context. Generally, they prefer, as do other elderly foreign workers, to remain in their cultural niches with their own religious institutions and migrant organizations. In comparison to the German population, the families of the first generation of migrants have more children and live more often in multigenerational households. Under the conditions of migration and in a foreign cultural context, the family fulfills a function of social-emotional support. Migrants possess a high potential for mobility. They commute between their homeland and the land of immigration. For some, this is a transitional strategy in order to reach a final decision, either to return to their homeland or to stay in Germany. For others, it is a long-term solution that enables them to make use of each country's respective advantages. PMID- 16362556 TI - [Strategies to improve the life situation of minority ethnic elders shown by means of the example of a model project in the Unna district]. AB - In Germany, the topic 'Ethnicity and Age' is a central concomitant of demographic and social change and has become a big challenge for existing structures of community care for elders. In this context, the Kreis Unna implemented a model project, which aimed at informing minority ethnic elders about the German care and welfare system. The project also tried to promote their usage of services and offers for elders in a sustainable way. Within the frame of the project, two information series were held. The first addressed minority ethnic elders and informed them about the German care and welfare system. The second qualified staff in the field of care and work for elders for the special needs of minority ethnic elders. The project was supported and evaluated by the Institute for Gerontology at the University of Dortmund. By viewing the results of the evaluation, it becomes evident that the two information and qualification series proved successful for both groups and, in general, are transferable to other municipalities. However, it also became clear that reducing barriers of access to the German care and welfare system is a very difficult process for both sides. A better usage of institutions and services of community care for elders by minority ethnic elders can only be accomplished by constant measures. PMID- 16362557 TI - [The employment of migrants in the field of care]. AB - This contribution tries to name key elements for a research agenda to study the increasing importance of the transnational migration for the formation of the social caring processes in the developed part of the world. First, the formal employment of people with migrant background in the German system for the delivery of care is regarded. Right now, there are no reliable data available which could give information about the degree of formal participation of migrants in the health and social care sector. It is agreed that in the future the service branch holds an increasing potential for employing people with migrant background. However, up to now, an initiative to qualify these younger migrant cohorts intensively is needed in order to compensate their educational and social disadvantages. The second part concentrates on the impact and development of informal work structures in care seen as irregular work participation in private households by migrants who are mostly from east-European countries. In a comparison between Germany, Austria and Italy, specific patterns of state policies toward this irregular work are analyzed; the phenomenon of a modern 'care drain' is analyzed and discussed in respect of the welfare state policies as well as of the potentials of action on the part of both migrants and family households. PMID- 16362558 TI - [Female geriatric patients with urinary incontinence symptoms and their control over pelvic floor muscles]. AB - Not only do anatomy and function of the pelvic floor play an important role as possible causes of female urinary incontinence, they are also crucial for its therapy. The aim of this case control study of female geriatric patients with symptoms of urinary incontinence was to determine the knowledge about their pelvic floor and to assess their ability to contract pelvic floor muscles voluntarily and reflexly. METHODS: A total of 377 female geriatric patients with symptoms of urinary incontinence were investigated in a Basis Assessment for Urinary incontinence. The ability to contract their pelvic floor muscles was examined by a digital vaginal palpation. The extent of the registered muscle strength was graded by the Modified Oxford Grading Scale by Laycock (1994). RESULTS: Of the patients, 65.5% were not aware of their pelvic floor and were not able to contract the pelvic floor muscles (Grade 0 to 1 by Laycock), 22% had an inaccurate knowledge and only performed an insufficient pelvic floor muscle contraction (Grade 2 by Laycock). Only 12.5% could contract their pelvic floor muscles properly (Grade 3 to 4 by Laycock). A subgroup of 83 patients had already absolved pelvic floor exercises in the past, 80 patients with conventional instructions, 3 patients with digital vaginal control. In this subgroup 54.2% of the patients were not able to contract the pelvic floor muscles (Grade 0 to 1 by Laycock) 25.3% only performed an insufficient contraction (Grade 2 by Laycock), while 20% were able to perform a sufficient and powerful contraction (Grade 3 to 4 by Laycock). The three patients in the past controlled by a digital vaginal palpation were part of this group and managed a pelvic floor muscle strength Grade 4 by Laycock. A high percentage of female geriatric patients with symptoms of urinary incontinence have a lack of understanding regarding the position and function of their pelvic floor. These results suggest that conventional pelvic floor muscle exercises without specific control are not an appropriate method to improve geriatric patients' ability to contract their pelvic floor muscles and to prevent urine leakage. PMID- 16362559 TI - [The emotional sensitivity of elderly people--validation of the Profile of Mood States for people over 60 years]. AB - The German short version of the "Profile of Mood States (POMS)" with 35 items (seven-point response scale; rating related to the past 24 hours) was psychometrically tested in a sample of elderly people of 60 years of age and older (n=690; 57% female). The internal structure of the POMS with four factors (dejection, fatigue, displeasure, vigour) could largely be replicated, even though the first three factors are intercorrelated. Reliability of the scales appears to be very good. The convergent validity of the POMS scales could be proved by correlations with the Beck Depression Inventory and the SF-36. Specific norms are presented for four age groups, men and women and elderly living in Eastern and Western Germany. So it seems appropriate to use the German short version of the POMS as a valid and economic instrument for gerontological studies. PMID- 16362566 TI - Assessment of MODIS-EVI, MODIS-NDVI and VEGETATION-NDVI composite data using agricultural measurements: an example at corn fields in western Mexico. AB - Although several types of satellite data provide temporal information of the land use at no cost, digital satellite data applications for agricultural studies are limited compared to applications for forest management. This study assessed the suitability of vegetation indices derived from the TERRA-Moderate Resolution Imaging Spectroradiometer (MODIS) sensor and SPOT-VEGETATION (VGT) sensor for identifying corn growth in western Mexico. Overall, the Normalized Difference Vegetation Index (NDVI) composites from the VGT sensor based on bi-directional compositing method produced vegetation information most closely resembling actual crop conditions. The NDVI composites from the MODIS sensor exhibited saturated signals starting 30 days after planting, but corresponded to green leaf senescence in April. The temporal NDVI composites from the VGT sensor based on the maximum value method had a maximum plateau for 80 days, which masked the important crop transformation from vegetative stage to reproductive stage. The Enhanced Vegetation Index (EVI) composites from the MODIS sensor reached a maximum plateau 40 days earlier than the occurrence of maximum leaf area index (LAI) and maximum intercepted fraction of photosynthetic active radiation (fPAR) derived from in-situ measurements. The results of this study showed that the 250 m resolution MODIS data did not provide more accurate vegetation information for corn growth description than the 500-m and 1000-m resolution MODIS data. PMID- 16362577 TI - Rib resection for live-donor nephrectomy. AB - INTRODUCTION: Living-related kidney transplants yield more favorable results than cadaveric kidney transplant. Although multiple techniques have been described for living-related donor nephrectomy, operation is generally performed subcostally in lateral decubitis position or by an 11th or 12th rib resection. Recently laparoscopic donor nephrectomy is getting popular. The aim of this study is to determine the rib resection increase the morbidity or not. MATERIALS AND METHODS: Between 1997 and 2004 in our center 118 living donor nephrectomies were performed. 15 of these patients did not come to follow-up controls. This study consists of 103 patients: 11th rib resection (30 patients) determined as group I, 12th rib resection (52 patients) determined as group II, subcostal incision (21 patients) determined as group III. All these three groups were compared with each other according to operation time, pleural or peritoneal defect, pneumothorax, blood transfusion, wound infection, length of hospital stay, postoperative analgesic requirement, return to threshold activities and incisional hernia. RESULTS: Patients whose 11th rib was removed had the shortest operation time. But pain due to surgery continued more than others in this group of patients. The risk of developing incisional hernia was seen most in patients who had subcostal incision. In this group of patients incidence of incisional hernia was 4 (19%). None of the patients had wound infection. We also did not experience any pneumothorax and blood transfusion requirement. Peritoneal or pleural opening occurred in 4 out of 103 patients accidentally and there was no difference between groups. There was also no difference between groups in terms of returning back to daily activation. CONCLUSION: Morbidity of nephrectomy done with removal of 12th rib was less compared with other groups. Resection of 11th should be reserved for patients with high residing kidneys and also for those with a polar artery of the upper pole. PMID- 16362578 TI - A rare case of an adult giant hydroureteronephrosis due to ureterovesical stricture presenting as a palpable abdominal mass. AB - The giant hydroureteronephrosis is a rare condition. We report a case of a unilateral giant hydroureteronephrosis in an adult patient presenting as palpable abdominal mass secondary to ureterovesical junction (UPJ) stricture. A 51-year old man presented with a 2-year history of a palpable abdominal mass. Physical examination revealed a grossly distended abdomen that the upper margin of distension was at the level of the epigastrium and the lower one at the suprapubic region. Abdominal ultrasound and computerized tomography demonstrated a very large cystic mass in the right side of retroperitoneum involving all the space from superiorly right sub-diaphragmatic area to the bladder inferiorly. The patient underwent exploration and a right giant hydroureteronephrosis as involving all the retroperitoneal space crossing midline to the left with a liquid content of approximately 7 l caused by stricture at the ureterovesical junction was seen. The hydronephrotic sac was thin and right nephroureterectomy was performed by opening the sac and draining-off the liquid. The literature was reviewed and the management of giant hydronephrosis was discussed. PMID- 16362579 TI - Extracorporeal shock wave lithotripsy in L-shaped kidneys: report of two cases. AB - The L-shaped or tandem kidney is a type of crossed ectopia with fusion in which the crossed kidney assumes a transverse position during its attachment to the inferior pole of the other kidney. Calculus formation is a well-known sequel of congenital abnormalities of the urinary tract and we like to present two patients harbouring calculi in L-shaped kidneys treated by extracorporeal shock wave lithotripsy (ESWL). The first patient had a calculus in renal pelvis of the transverse kidney. The second case had multiple calculi in renal pelvis and calices of the vertically positioned kidney. After three sessions, first patient was stone free, whereas no fragmentation in the second case. As a conclusion, ESWL might be an option for L-shaped kidneys. PMID- 16362580 TI - Case report: octreotide as an adjunct to embolisation in the management of recurrent bleeding upper gastrointestinal metastases from primary renal cell cancer. AB - The recommended treatment for patients with severe bleeding from upper gastrointestinal (GI) metastases is embolisation. We report a case in which despite adequate embolisation major haemorrhage from renal cell carcinoma (RCC) gastric metastases continually recurred. During a severe bleed refractory to embolisation octreotide was used to control and prevent further bleeding. No further episodes of severe haemorrhage occurred over the following 23 months since starting treatment. Octreotide has been observed to be effective both in the acute management of persistent haemorrhage and in prevention of subsequent haematemesis from GI metastatic RCC. PMID- 16362581 TI - Dribbling of urine due to ectopic vaginal insertion of an upper pole ureter diagnosed by magnetic resonance urography. AB - Imaging the urinary tract in children has been mainly based on ultrasound (US) and intravenous pyelography (IVP), but these imaging modalities rarely show ectopic insertion of the ureter and the condition is often under diagnosed due to unusual clinical presentations. Recently newer magnetic resonance urography sequences have been developed that provide better delineation of the urinary tract. We report a 3-year-old girl presenting continuous dribbling of urine in which magnetic resonance urography revealed ectopic vaginal insertion of the ureter. PMID- 16362582 TI - A giant incisional flank hernia. PMID- 16362583 TI - The relationship between the perineal length measurements and the urodynamic stress incontinence. AB - OBJECTIVE: The purpose of this study was to assess the relationship between the length of perineal body, the anal position index, the total fourchette-coccyx distance and the anal-coccyx length and female urodynamic stress urinary incontinence (USUI). PATIENTS AND METHODS: The study included 57 women with USUI (group A) and 45 women without USUI as control (group B). All patients underwent a urodynamic evaluation and measurement of the length of perineal body (PB) and the distance between the fourchette and the inferior margin of coccyx (FC). PB is the distance between the fourchette and the center of anal orifice. The ratio PB/FC is the anal position index. Subtracting the PB length from FC distance equals the anal-coccyx (AC) length. RESULTS: The student's t-test showed no significant statistical difference between any of the measured lengths in the two groups. Moreover, in the USUI group, ANOVA did not show any significant relationship between the urodynamic parameters and the measured lengths. Regarding to the investigated urodynamic parameters only the maximum urethral pressure (Pura max) presented a significant statistical difference between the two groups (P = 0.009). CONCLUSION: Theoretically, a differentiation of perineal distances probably brings to the surface a laxity of the pelvic floor, a factor predisposing to USUI. Furthermore, the anterior position of anus can be a cause of constipation which also contributes to USUI. However, our study did not reveal any relationship between the perineal length measurements and USUI. Further investigation with longer series of patients is needed to show if this simple, inexpensive and non-invasive test could be added in the armamentarium of the diagnostic and prognostic investigation of USUI. Regarding to the urodynamic parameters, our results emphasize the importance of the Pura max parameter in the urodynamic evaluation of USUI. PMID- 16362584 TI - Case series: pseudotumor formation in bladder as a late complication of pelvic and inguinal surgeries. AB - Inguinal and pelvic surgeries are rarely complicated by urologic complications. Eleven cases having such history admitted to our department with urologic symptoms were shown to have microscopic hematuria and bladder lesions as well. All of them were explored some due to retained gauze compress some due to suspected malignant formations on computerized tomography. Furthermore all were shown to be benign in nature. The cases were discussed under scope of current literature. PMID- 16362585 TI - Case report: cystitis glandularis mimics bladder tumour: a case report and diagnostic characteristics. AB - We report a rare case of a 26-year-old patient presented with 3-week history of frank haematuria and suprapubic discomfort. Investigations revealed a tumour arising from the bladder wall and histology demonstrated cystitis glandularis. The literature review highlights the rarity of cystitis glandularis presented in such manner. Our radiographic and endoscopic images can also assist future diagnosis of the condition especially in patients in such age groups. PMID- 16362586 TI - Urinary retention secondary to presacral myelolipoma; first reported case diagnosed by prostate TUR and requiring a cystectomy. AB - Obstructive voiding symptoms may exceptionally be caused by extrinsic compression. We herein present a singular case of a 68-year-old male that presented with urinary retention and underwent prostate trans-urethral resection (TUR) with histology showing benign prostatic hyperplasia admixed with large amounts of myelolipoma tissue. To the best of our knowledge this is the first reported presacral myelolipoma diagnosed at prostate trans-urethral resection (TUR). Computed tomography revealed a 13 x 9 cm presacral mass displacing the rectum. Even though myelolipomas are regarded as benign, this case behaved aggressively since compressive effect evolved to severe constipation and eventually required a cystectomy. PMID- 16362587 TI - Utility of volume adjusted prostate specific antigen density in the diagnosis of prostate cancer in Arab men. AB - BACKGROUND: This study was undertaken to assess the utility of prostate specific antigen (PSA) and PSA density (PSAD) in discriminating between benign and malignant prostate disease in the Kuwaiti Arab population. METHODS: A total of 100 consecutive patients suspected of having prostate cancer because of serum PSA > 4 ng/ml, or detection of a prostatic nodule on rectal examination were further investigated by determination of PSAD, TRUS of prostate, sexant prostatic biopsy and histological analysis to establish the correct diagnosis. Other diagnostic measures included the determination of the area under the receiver operating characteristic (ROC) curve, sensitivity and specificity. RESULTS: Of the 100 prostate biopsies that were performed, 33 cases were confirmed to be prostate cancer and 67 were described as benign lesions comprising benign prostatic hyperplasia (BPH) with or without prostatitis. The age range for patients with prostate cancer was 42-90 years, and 52-90 years for those without prostate cancer. The mean prostate volume was 58.82 cc (range 9-177 cc) and 62.60 cc (range 15-140 cc), the mean PSA value was 36.65 ng/ml (range 5.8-200 ng/ml) and 16.49 ng/ml (range 1.4-46.0 ng/ml), while the mean PSAD was 0.92 (range 0.046 5.714) and 0.452 (range 0.034-2.294) for patients with prostate cancer and patients without prostate cancer respectively. Patients with PSA less than 4 ng/ml (3 cases) all had benign prostate lesions, and 7 cases with PSA more than 50 ng/ml all had prostate cancer and were excluded because values above 50 ng/ml have close to 100% specificity for prostate cancer. Further analysis was done on the remaining 90 cases which were patients with a PSA between 4 and 50 ng/ml. The discriminating power of serum PSA for detecting prostate cancer as estimated by the area under ROC was 0.686 while that for PSAD was 0.732. The maximum likelihood for a positive PSA was at a PSAD cut-off point of 0.32. For the PSA cut-off point of l0 ng/ml, the sensitivity was 80%, and specificity was 42.2%. For the PSAD cut-off point of 0.32, the sensitivity was 58% and the specificity 76.6%. CONCLUSIONS: Determination of PSAD is not a useful adjunct to serum PSA values in the range of 10-50 ng/ ml in our population. PSAD value less than 0.32 with PSA less than l0 ng/ml strongly suggests benign disease. PMID- 16362588 TI - Cerebrospinal fluid prostate specific antigen (CSF PSA) in prostate cancer patients with lower spine metastasis. AB - AIM: In this prospective study, our aim was to investigate the CSF PSA levels and CSF/Serum PSA ratios in patients with prostate cancer with lower spine metastasis. METHODS: The study involved patients with prostate cancer (n = 15), benign prostatic hyperplasia (n = 17) and non-prostatic disease (n = 9). Serum and CSF were obtained prior to spinal anesthesia for urological surgery. Total PSA levels in the serum and CSF were measured by electrochemiluminescence immunoassay. The results were tested statistically using the Mann-Whitney U test. RESULTS: The mean serum PSA levels were 20.36 ng/ml in the prostate cancer patients, 5.37 ng/ml in the BPH patients and 0.76 ng/ml non-prostatic disease. The mean CSF PSA levels in groups were 0.127, 0.051 and 0.027 ng/ml, respectively. The mean CSF PSA/serum PSA ratios in groups were 0.007, 0.018 and 0.042, respectively. This result is statistically significant (P < 0.001). CONCLUSIONS: Although mean serum PSA and CSF PSA levels in the patients with cancer of the prostate and lower spine metastasis are higher than those in the others, the mean CSF PSA/serum PSA ratio is lower. However, clinical usefulness of CSF PSA value and CSF PSA/ Serum PSA ratio can be limited because CSF PSA values are usually very low, and CSF PSA/Serum PSA ratio of 4 prostate cancer patients are as high as 1 BPH patient. PMID- 16362589 TI - Case report: angioleiomyoma of the spermatic cord: a rare scrotal mass. AB - We report a case of angioleiomyoma of the spermatic cord. Tumours arising from the spermatic cord are rare and most of them are benign. The commonest are lipomas with sarcomas being the predominant malignant variety. This is the first reported case of angioleiomyoma of the spermatic cord. PMID- 16362590 TI - Coagulation parameters in the patients with Fournier's Gangrene. AB - AIM: In this prospective study, we aimed to determine the coagulation parameters in the patients with Fournier's gangrene. METHODS: The study group is consisted of 12 consecutive Fournier's gangrene patients (11 men and 1 woman) and control group is consisted of patients with periurethral abscess (n = 2), scrotal abscess (n = 4) and epididimorchitis (n = 4). Fibrinogen, protein C, protein S, antithrombin III, lupus anticoagulant, cardiolipin IgG and IgM, prothrombin time international normalized ratio, activated partial thromboplastin time, platelet count, proaccelerin, antihemophilic globulin (FVIII), albumin, and calcium were evaluated in all subjects. Tissue specimens were taken from Fournier's gangrene patients. These specimens were tested for arterial and venous thrombosis using light microscopy. RESULTS: All of the patients with Fournier's gangrene had both arterial and venous thrombosis in tissue specimens. The levels of fibrinogen and FVIII were high, the level of protein C was low in 12 patients. Lupus Anticoagulant was positive 11 of 12 patients. CONCLUSIONS: According to our findings, we think that some coagulation parameters (FVIII, Lupus anticoagulant, protein C, fibrinogen) may be diagnostic for Fournier's gangrene. PMID- 16362591 TI - 3-Dimensional computerized tomography in follow-up of patients with urinary diversion. AB - INTRODUCTION: The aim of our study was to evaluate the usefulness of 3 dimensional computerized tomography (3D-CT) in routine follow-up of patients who had undergone radical cystectomy and different kinds of urinary diversions and compare it with conventional CT. PATIENTS AND METHODS: Nineteen patients (18 men, 1 woman) who had undergone radical cystectomy and different kinds of urinary diversions with diagnosis of invasive bladder cancer were enrolled into the study. The mean age of the patients was 55.5 (46-69) years. For all patients, conventional CT was performed, followed by 3D reconstruction of these images. RESULTS: Orthotopic ileal neobladder (Abol-Enein and Ghoneim procedure) was performed in 12, rectosigmoid pouch (Mainz pouch II) in 2 and ileal conduit in 5 patients. There were no pathological findings visible either on conventional CT or on 3D-CT, but the shape, configuration, and the relationships of the pouch with ureters, urethra and other adjacent organs were much better visualized on 3D CT. CONCLUSION: 3D-CT did not have any advantages over conventional CT for showing pathological findings that were oncological origin, but as the anatomy of the lower urinary tract can be perfectly demonstrated, we think that it may be used for planning of a re-operation after urinary diversion and may help the urologist who has less experience with radiological studies to evaluate pouch configuration and indentations to the adjacent organs better. PMID- 16362592 TI - Extensively cystic renal neoplasms in adults (Bosniak classification II or III)- possible "common" histological diagnoses: multilocular cystic renal cell carcinoma, cystic nephroma, and mixed epithelial and stromal tumor of the kidney. AB - OBJECTIVE(S): To give an algorithm for resolution of extensively cystic renal neoplasms, preoperatively classified in the Bosniak classification as a category II and III. METHODS: From 1991 to 6/2004, 701 patients with 727 renal tumours were surgically treated at our hospital. Extensively cystic tumours were found in 10 cases. Extensively cystic tumours were defined as multicystic tumours without any solid nodules visible neither on CT, nor grossly in the specimen at operation (the Bosniak classification type II or III). RESULTS: Seven multilocular cystic renal cell carcinomas, three mixed epithelial and stromal tumour of the kidney and one cystic nephroma were diagnosed on histology. CONCLUSION(S): Extensively cystic renal tumours classified as the Bosniak type II or III correspond histologically to the entities mentioned above (multilocular cystic renal cell carcinoma, cystic nephroma, mixed epithelial and stromal tumour of the kidney). These entities cannot be distinguished one from another on preoperative imaging studies. A preoperative biopsy and intra-operative frozen-section analysis do not lead to a correct diagnosis in many cases. Fortunately, the operative strategy is the same for all these tumours. In such cases, the nephron sparing surgery is indicated, whenever technically feasible, as almost all extensively cystic renal tumours have a good prognosis. PMID- 16362593 TI - The role of urine osmolality and ions in the pathogenesis of primary enuresis nocturna and in the prediction of responses to desmopressin and conditioning therapies. AB - Aim of the study was to determine the role of nocturnal and daytime urine volume, osmolality and ion excretions in the pathogenesis of primary monosymptomatic enuresis nocturna (PMEN) and in the prediction of response to desmopressin and conditioning therapies. Fifty-five children with PMEN between the ages 5 and 15 years were included to the study. The patients were randomly divided into three groups Group 1: Twenty enuretics having intranasal desmopressin 1x 20 microg treatment for 2 months Group 2: Twenty enuretics having conditioning therapy for 2 months. Group 3: Fifteen enuretics having intranasal izotonic solutions as placebo. The control group consisted of 15 healthy children. Urine osmolality, sodium, potassium, chloride, magnesium and creatinine levels were investigated in both daytime and nighttime urines. Fractional sodium, potassium, magnesium, chloride excretions were calculated. Wilcoxon, Mann-Whitney U, Kruskal-Wallis, Chi-square, Student's t and Pearson correlation tests were performed. The ratio of night/daytime urine osmolality was significantly decreased in enuretic children. In addition, the ratio of night/daytime urine Cl and K excretions were also significantly decreased in enuretics. Response rate to desmopressin and conditioning treatments were statistically higher than placebo control. The difference between response rates of desmopressin and conditioning therapies was not found statistically significant. Pretreatment values of urine volume osmolality and ions were not observed as predictive factors in response to desmopressin or conditioning therapy. In conclusion, nightly decreased potassium and chloride excretions were found to have a role in the pathogenesis of primary enuresis nocturna. Urine volume, osmolality and ion excretions are not suggested to be used in the prediction of response to desmopressin and conditioning therapies. PMID- 16362594 TI - Testicular choriocarcinoma metastatic to the skin. AB - Pure testicular choriocarcinoma with metastasis to the skin is a very rarely seen phenomenon. Choriocarcinoma has a distinct propensity for early hematogenous spread to distant sites. We present the case of a 42-year-old white male with testicular choriocarcinoma with skin metastasis. PMID- 16362595 TI - Routine antibiotic prophylaxis is not necessary for no scalpel vasectomy. AB - No scalpel vasectomy (NSV) is the recommended technique for male contraception. Majority of surgeons use oral antibiotics for 3-5 days following the procedure. The reasons cited for routine use of antibiotics include that most of these operations are performed in camps where asepsis in less than adequate, most patients are unclean and unhygienic surrounding environment. However, there is no database of wound infection following NSV. This study was undertaken to find out the incidence of wound infection following No scalpel vasectomy if no antibiotic prophylaxis is used. Patients undergoing NSV in the minor operation theatre attached to the Surgical Out Patient Department were included in the study. A standard preoperative part preparation protocol was followed in all patients. All procedures were performed under local anesthesia after obtaining informed consent. Post-operatively patients were advised an oral anti-inflammatory drug for 3-5 days and were reviewed after 1 week for development of wound infection. Over a 4-year period, 322 patients underwent NSV and no patient developed wound infection. Hence, it is concluded that routine antibiotic prophylaxis is not necessary for NSV. PMID- 16362596 TI - Clinical management of large adrenal cystic lesions. AB - The widespread use of ultrasonography and computed tomography has resulted in an increased diagnosis of large sized adrenal cysts with diameters of more than 5 cm. Most of these adrenal cystic lesions are clinically silent and are therefore often diagnosed incidentally. Since up to 7% of adrenal cysts are malignant, a careful hormonal, morpho-functional and instrumental evaluation is mandatory. In particular, functioning adrenal carcinomas or pheochromocytomas have to be ruled out. Fine needle aspiration cytology as well as examination of a punch biopsy specimen of the cystic wall are of limited value, as there is considerable overlap in cytologic and histologic features of benign and malignant adrenal cystic lesions. Immediate surgical excision is indicated in the presence of symptoms, suspicion of malignancy, increase in the size or detection of a functioning adrenal cyst. En bloc adrenalectomy, preferably by a laparoscopic approach, has become the treatment of choice. PMID- 16362597 TI - Terazosin in the treatment of premature ejaculation: a short-term follow-up. AB - AIM: The aim of the present study was to evaluate the efficacy of terazosine in patients with premature ejaculation and lower urinary tract symptoms (LUTS), after excluding other sexual disorders and chronic prostatitis. METHODS: A total of 90 patients with premature ejaculation and LUTS were enrolled to the study after excluding sexual disorders, prostatitis and benign prostatic hyperplasia. The patients were divided into two groups. Sixty patients in group 1 were treated with terazosine 5 mg daily for a month. Patients were followed monthly and questioned for their ejaculation problem. The results were classified as cure, improvement and ineffective. If patients showed improvement and ineffectiveness, the treatment was continued with 10 mg daily for the following month. Group 2 was included 30 patients, and placebo was applied for a month. At the end of this period, in patients who did not show any improvement, terazosine 10 mg was started. RESULTS: In the treatment group, at the 1st month follow-up, 21 patients (35%) were cured, 20 (33.3%) showed improvement. In 19 (31.7%) patients, the treatment was ineffective. In group 2, 9 (30%) patients showed improvement and the rest had no-changes after one-month follow up. There was statistically significant difference between two groups (Pearson chi2 test = 0.000). Later, terazosine 10 mg was given to the patients in group 2 and to the patients who showed improvement or unsuccessful result with terazosine 5 mg. With terazosine 10 mg, 10 (14.5%) patients were cured, 29 (42.2%) patients were improved. Finally, terazosine treatment in patients with premature ejaculation was found to be effective in 60 patients (66.7%). CONCLUSION: Alpha blockers seem to be physiological medical agents in the treatment of premature ejaculation since ejaculation is under sympathetic control. Moreover, these agents are effective in lower urinary tract and they should be used in patients with premature ejaculation and lower urinary tract symptoms. PMID- 16362598 TI - Correspondence Re: "Garat JM, de la Pena E, Caffaratti J, Villavicencio H. Prevention of vesicoureteral reflux at the time of complete primary repair of the exstrophy-epispadias complex. Int Urol Nephrol. 2004; 36: 211-2". PMID- 16362599 TI - Curvularia geniculata fungal peritonitis: a case report with review of literature. AB - Peritonitis is the "Achilles heel" of patients on peritoneal dialysis. Fungal peritonitis though not that common is associated with increased morbidity and mortality. We report the first case in the literature of a patient who developed peritonitis caused by Curvularia geniculata. We discuss the reported cases of Curvularia lunata peritonitis in literature and emphasize the early removal of catheter and treatment with intravenous amphotericin B. PMID- 16362600 TI - Hemodynamic effects of sildenafil citrate (Viagra) on segmental branches of bilateral renal arteries. AB - INTRODUCTION: Erectile dysfunction (ED) is one of the most encountered male health problem and sildenafil citrate is used worldwide for treatment of ED. Aim of the present study was to determine and evaluate hemodynamic effects of sildenafil citrate on segmental branches of bilateral renal arteries of cases with ED. PATIENTS AND METHODS: Total of 35 male patients aging between 34 and 56 years were recruited into study. Patients were evaluated with renal color Doppler sonography (CDS) before and after (45-75 min) peroral sildenafil citrate (50 mg) administration. Peak systolic flow rate (V(max)), end diastolic flow rate (V(min)), resistive index (RI) and pulsatility index (PI) of renal artery segmental branches were determined before and after sildenafil citrate administration. RESULTS: V(min) measured at right lower segment was significantly reduced after sildenafil administration (p = 0.030). Accordingly mean RI and PI of right lower segment of renal artery slightly increased after drug administration (p = 0.007 and p = 0.012, respectively). The measurements on other segments had no significant change after sildenafil treatment when compared to baseline values. On this single dose administration no side effect was recorded. CONCLUSION: Findings of the present study demonstrated that peroral sildenafil citrate usage had slight effects on hemodynamic parameters of lower segmental branch of right renal artery. This result could not be interpreted as robust evidence for hemodynamic changes of renal arteries associated with sildenafil usage nevertheless warrants more comprehensive research with larger number of patients. PMID- 16362601 TI - A rare case of acute renal infarction due to atrial fibrillation mimicking renal colic. AB - A 70-year-old patient with acute renal infarction due to chronic atrial fibrillation is presented. The clinical presentation of the patient was suggestive of renal colic. Computerized tomography was consistent with acute renal infarction and confirmed the diagnosis. After giving anticoagulation and antiarrhythmic treatment, she was discharged with clinical improvement. High clinical suspicion is necessary on an old patient who has thromboembolic risk factors with the complaint of abrupt-onset flank pain. PMID- 16362602 TI - Detection of occult renovascular disease in unexplained chronic kidney disease. AB - Atherosclerotic renal artery stenosis (RAS) is a recognized cause of renal impairment. RAS is often overlooked in unexplained chronic kidney disease (CKD). A retrospective analysis of renal angiograms was performed to determine the prevalence of occult renovascular disease in 64 (M:F, 46:18; ages 21-81 years [median 60 years]) patients with unexplained CKD. Twelve patients had diabetes mellitus (type II: 11) and 43 were smokers. Median serum creatinine was 5.2 mg/dl (range 1.5-10.6 mg/dl). Group A included patients with unexplained CKD and with no risk factors for RAS and Group B had patients with increased risk for RAS. A narrowing of the renal vessel, main artery or branch, by >50% on renal arteriography was used as diagnostic criteria for RAS. 31/64 patients had positive angiographic findings. Thirteen patients had unilateral RAS, 9 had bilateral RAS, 5 had unilateral stenosis with occlusion on the opposite side, 3 had unilateral occlusion and 1 had a solitary kidney with RAS. 19/34 (54%) in Group A and 12/30 (40%) in Group B had a positive renal angiogram. In 10 patients with a rise in serum creatinine on recent introduction of ACE inhibition, 2 had evidence of RAS on renal arteriography. Eleven patients underwent angioplasty and 2 reconstructive surgeries. In 4 patients, blood pressure control improved and anti-hypertensive drug requirements were reduced, whilst renal replacement therapy was postponed in 4, by 2-24 months. In 18 patients, the lesions were not amenable to angioplasty or reconstructive surgery. Four patients did not benefit in any form with intervention. Occult atheromatous renal vascular disease is a common, not readily predictable and potentially correctable etiology of unexplained CKD. PMID- 16362603 TI - Prostate cancer in the elderly. AB - Prostate cancer is the second leading cause of cancer deaths among men. Despite earlier diagnosis due to prostate specific antigen (PSA) screening, it is still a disease of the elderly. Diagnosis is based on digital rectal examination (DRE) and PSA assessment. Refinements in PSA testing (age-specific reference ranges, free PSA, PSA density and velocity) increased specificity and limited unnecessary prostate biopsies. Diagnosis in earlier stages (T1 and T2) commonly leads to cure with current treatment modalities. These include radical prostatectomy, external beam radiotherapy and brachytherapy. Other treatment options under development include cryotherapy and high-intensity focused ultrasound. Metastatic prostate cancer is incurable and treatment is based on hormonal therapy. Cytotoxic chemotherapy has only limited role in hormone-independent prostate cancer. Radioisotopes and biphosphonates may alleviate bone pain and prevent osteoporosis and pathological fractures. Follow-up is based on PSA. Prognostic factors for recurrence include stage, Gleason score, pre- and posttreatment PSA. Quality of life issues play an important role in selecting treatment, especially in the elderly due to comorbidities that may negatively affect the overall quality of life. A holistic approach is recommended addressing all quality of life issues without focus only in cancer control. PMID- 16362605 TI - Bordetella avium infection in chickens and quail in Nigeria: preliminary investigations. PMID- 16362606 TI - Effect of monensin on serum lipoproteins, triglycerides, cholesterol and total lipids of periparturient dairy cows. AB - Serum concentrations of very low-density lipoproteins (VLDL), low-density lipoproteins (LDL), high-density lipoproteins (HDL), triglycerides, cholesterol, and total lipids of a group of monensin-treated cows (n = 7) were compared with those of a control group (n = 6) from about 10 days before calving to 45 days postpartum. Monensin was fed in the diet from about 20 days before predicted calving date to 45 days after parturition. Blood samples were obtained on days 20 and 10 before predicted calving, and on days 4, 15, 25 (phase I), and days 35 and 45 (phase II) post calving. Cholesterol (p = 0.046) and total lipids (p = 0.003) were lower in the treatment group 10 days before calving. Concentrations of serum triglycerides and VLDL (p = 0.017), and that of LDL (p = 0.032) were significantly higher in the treatment group in phase I postpartum. No significant difference was detected between groups during phase II. Comparison of the entire length of the trial showed higher concentrations of triglycerides and VLDL in the treatment group (p = 0.009). It is concluded that monensin is effective in increasing secretion of triglyceride-rich lipoproteins from the liver during the postpartum transition period of dairy cows. This may be effective in reducing the intensity of hepatic lipidosis and the incidence of periparturient diseases resulting from disturbances in energy metabolism. PMID- 16362607 TI - Effect of natural antioxidants on superoxide dismutase and glutathione peroxidase mRNA expression in leukocytes from periparturient dairy cows. AB - During the peripartum period, high-yielding dairy cows experience metabolic stress, which alters their homeostasis and exposes the cows to illness. The aim of this study was to quantify the expression levels of genes involved in antioxidant defences during the transition period in the blood of dairy cows and to evaluate the regulative activity on these genes of natural antioxidants in the diet. Three groups of 7 heifers each, at the 7th month of pregnancy, were used. Starting from 3 weeks before the expected calving date (-22 days), the three groups were allotted to the following experimental treatments: control (CTR, basal diet); lycopene (LYC, basal diet + lycopene 540 mg/day) and grape polyphenols (POL, basal diet + grape polyphenols 10 g/day). Blood was sampled at 22 and 8 days before and 8, 15 and 22 after calving and analysed for the expression level of glutathione peroxidase (GPx) and superoxide dismutase (Cu/ZnSOD) using the real-time PCR technique with LUX (Light Upon eXtension) fluorogenic primers. During the peripartum period (-22 days until + 22 days from calving), Cu/ ZnSOD mRNA expression decreased (p<0.05) in the CTR and LYC groups, but increased at 15 days after calving in the POL group. No significant differences were found in GPx mRNA expression. The results suggest that grape polyphenols may have a controlling effect on peripartum metabolic stress through modulation of superoxide dismutase expression. PMID- 16362608 TI - Comparison of activities of enzymes related to energy metabolism in peripheral leukocytes and livers between Holstein dairy cows and ICR mice. AB - Activities of enzymes related to energy metabolism and isoenzyme patterns of lactate dehydrogenase (LDH) were determined in peripheral leukocytes and livers of Holstein dairy cows and Institute of Cancer Research (ICR) mice. In dairy cow liver, activities of enzymes in glycolysis, malate-aspartate shuttle and lipogenesis were lower, but activities of glucose-6-phosphatase in gluconeogenesis were higher than those in mouse liver. Glucokinase activities were below detection limit in leukocytes and liver of the cows. Dairy cow leukocytes and liver showed the isoenzyme patterns with dominance of LDH-1, -2 and-3, whereas mouse leukocytes and liver showed that LDH-5 was dominant. The LDH isoenzyme patterns were very similar between leukocytes and liver in each animal species. Some enzymes in leukocytes may reflect those enzymes activities in liver and be a useful indicator for energy metabolism in animals. PMID- 16362609 TI - Periodic release of gastrointestinal helminth eggs in native chickens from Dschang in the western highlands of Cameroon. PMID- 16362610 TI - Cross-sectional prevalence of helminth infections in cattle on traditional, small scale and large-scale dairy farms in Iringa district, Tanzania. AB - A cross-sectional study was carried out to determine the prevalence of gastrointestinal (GI) nematodes and flukes (Fasciola and amphistomes) infection in communally grazed traditional cattle, zero-grazed small-scale dairy cattle and intensively grazed large-scale dairy cattle through examination of helminth eggs in faeces. Results indicated that the type of management, especially the grazing habit, has a significant influence on the prevalence and intensity of GI nematodes and flukes. The prevalence of GI nematodes in traditional, large-scale dairy and small-scale dairy cattle was 67%, 44.4% and 37%, respectively, with the highest faecal egg counts in calves. The overall prevalence of Fasciola gigantica in traditional, large-scale dairy and small-scale dairy cattle was 63.8%, 46.2% and 28.4%, respectively. The prevalence of amphistomes was 81.9%, 55.5% and 41.1% in traditional, large-scale dairy and small-scale dairy cattle, respectively. The high prevalence of flukes in the traditional system was attributed to communal grazing and watering management practices. Stomach flukes recovered in examined cattle at the abattoir were Calicophoron microbothrium and Cotylophoron jacksoni. About 42.1% of infected animals had both Fasciola and amphistomes. The prevalence of both GI nematodes and flukes varied greatly among villages and farms. The prevalence of both Fasciola and amphistomes was higher in adults (58.5%, 75.2%) than in yearlings (36.5%, 51.5%) or calves (24.9%, 47.2%). The variation in the prevalence of both GI nematodes and flukes among management and age groups within systems can be used as an entry point towards rational use of anthelmintics for each management system. More studies on seasonal transmission pattern of all these parasites are required in order to design rational, economic and locally sustainable parasite control programmes. PMID- 16362611 TI - Expression of putative Escherichia coli heat-labile enterotoxin (LT) receptors on intestinal brush borders from pigs of different ages. AB - Many strains of enterotoxigenic Escherichia coli (ETEC) that cause diarrhoea in young piglets secrete a heat-labile enterotoxin (LTp) that binds to specific glycoconjugates on porcine intestinal epithelial cells. Binding of LTp to an appropriate glycoconjugate facilitates the uptake and trafficking of the toxin into the cell, where it stimulates intracellular changes that promote fluid secretion and diarrhoea. The objective of the current study was to identify the LTp-binding glycoconjugates on porcine intestinal epithelial cells, the natural target cells for LTp. We found that LTp binds, in an age-correlated manner, to an acidic glycosphingolipid (GSL) that co-migrated with GM1 on thin-layer chromatography (TLC), a small acidic GSL that appears to be a sulphatide, a neutral GSL that co-migrated with neolactotetraglycosylceramide (nLc4) on TLC, and two glycoproteins (36 and 205 kDa). Of these potential LTp receptors, the GM1 co-migrating GSL was detected most intensely in young animals, while the other four LTp-binding glycoconjugates were detected most intensely in older pigs (> or= 4 weeks). Since ETEC primarily cause disease in young piglets, the GM1-co migrating GSL is the most likely candidate for a functional LTp receptor. PMID- 16362612 TI - Pharmacokinetics of flunixin after intravenous administration in healthy and endotoxaemic rabbits. AB - The pharmacokinetics of flunixin were determined after intravenous bolus injection at a single dose (2.2 mg/kg) in healthy rabbits and diseased rabbits with Escherichia coli lipopolysaccharide-induced septic shock. Six adult New Zealand White rabbits were used. Concentrations of drug in plasma were determined by HPLC. Pharmacokinetics were best described by a two-compartment open model. In healthy rabbits, there was a high plasma clearance (0.62 L/(h kg)), and a relatively short elimination half-life (1.19 h). In endotoxaemic rabbits, total plasma clearance (0.43 L/(h kg)) was significantly lower (p<0.05), and elimination half-life (1.90 h) and AUC(0-infinity) (5.29 (microg h)/ml) were significantly higher (p<0.05) than in healthy animals. The changes of pharmacokinetics of flunixin in rabbits with septic shock could be of clinical significance, and may require monitoring of plasma flunixin levels in endotoxaemic status. PMID- 16362613 TI - Relationship between welfare and udder health indicators in dairy ewes. AB - The relationships between welfare and udder health indicators were investigated in 120 midlactation Comisana ewes over a 90-day period. The welfare indicators monitored were the humoral response to chicken egg albumin (OVA), the cell mediated immune response to phytohemagglutinin, and the plasma cortisol levels. Changes in milk components, in somatic cell count (SCC) and in bacterial count, in leukocyte populations and in anti-OVA IgG concentration were used to monitor the ewe udder health. Correlations between welfare and udder health indicators were then tested. The skinfold thickness, representative of the cell-mediated immune response, was correlated positively with the percentage of neutrophils in milk and negatively with the chloride content, the percentage of lymphocytes and anti-OVA IgG concentrations in milk. The plasma antibody titres to OVA, representative of the humoral immune response, correlated positively with the anti-OVA IgG levels in milk and negatively with the lactose content and to the percentage of neutrophils. Plasma cortisol levels correlated positively with the chloride content in milk. Our study demonstrates that information about the immune condition of animals can be successfully achieved by monitoring the immunological responses of the mammary gland. The detection of reliable indicators of animal welfare in milk can help in reducing the stress from animal handling and blood sampling from lactating animals. PMID- 16362614 TI - Protection of the reproductive tract of young chicks by Newcastle disease virus induced haemagglutinationinhibition antibodies. AB - The present study was conducted to assess the haemagglutination-inhibition (HI) titres required to protect the chicken reproductive tract against direct damage caused by Newcastle disease virus (NDV). Precociously induced oviduct and uterus by oestrogen treatment of young chicks were used to assess the damage or protection against the damage by analysis of ciliostasis or histopathological lesions. Unvaccinated day-old female white leghorn chickens were used as the maternally derived antibody (MDA) group. Chickens were vaccinated with either a live lentogenic vaccine on day 14 of age or, along with it, an inactivated vaccine at day 36 of age, to generate birds with a range of primary or secondary response induced HI antibodies. Birds with different HI antibody levels were challenged with virulent NDV. It was found that a HI antibody titre of 128 and above was protective against direct damage of the reproductive tract, while the 32-64 titre range was protective when derived through secondary vaccination only. PMID- 16362615 TI - Marmoset B95a cells: a sensitive system for cultivation of Peste des petits ruminants (PPR) virus. PMID- 16362616 TI - Dangerous personalities. PMID- 16362617 TI - Differences in students' perceptions of learning compulsory foundation biochemistry in the health sciences professions. AB - Student approaches to learning vary from surface approaches to meaningful, deep learning practices. Differences in approach may be related to students' conceptions of the subject, perceptions of the learning environment, prior study experiences and performance on assessment. This study aims to explore entering students' conceptions of the unit they are about to study and how they intend to approach their studies. It involved a survey of 203 (of 250) first year students in a cross disciplinary unit in the Faculty of Health Sciences. They were asked to complete an open-ended response survey, including questions on what they thought they needed to do to learn biochemistry and what they thought the study of biochemistry was about. A phenomenographic methodology was used to identify categories of description for the questions. The paper will describe the categories in detail, the structural relationship between the categories and the distribution of responses within categories. The study reports a relationship between conception of the topic and approaches to learning. Students with more complex and coherent conceptions of the topic report that they were more likely to adopt deeper approaches to study than those with more fragmented conceptions. However, compared to previous studies, a surprisingly high proportion of students with more cohesive conceptions still intended to adopt more surface approaches. This may reflect the particular context of their learning, namely in a compulsory unit involving material for which most students have minimal background and difficulty seeing its relevance. Implications for teaching such foundation material are discussed. PMID- 16362618 TI - Students' reactions to three typical examinations in health sciences. AB - OBJECTIVE: A search for universal and particular changes in emotional, behavioral and cognitive assessments in relation to three types of examinations: an oral presentation, an objective structured clinical examination (OSCE) and a pencil and paper examination. METHODS: One hundred and two students of health professions completed the Profile of Mood States (POMS) questionnaire before (t1) and after (t2) each type of examination. Data regarding stress-related symptoms, attitudes, and preparation behavior were collected at (t2). RESULTS: POMS six subscales scores at (t1) did not differ by examination type and five of them were statistically significant higher at (t1) than (t2) regardless of examination type. "Preparing behavior" also emerged as a universal feature. As for the particular aspects of each examination, OSCE students felt more depression dejection and fatigue at (t2) than at (t1). Oral presentation was perceived as the most difficult by students, who also reported more symptoms. For this type of examination, the students regained their confidence towards the end of the presentation and perceived classmates as most supportive and least disturbing. The pencil and paper examination was evaluated as the easiest by students, produced fewest symptoms and no excitement during the examination. CONCLUSIONS: Three types of examinations, frequently used in health sciences, appeared to initiate different physical, emotional, and social reactions. PMID- 16362619 TI - Medical students' mindset for reflective learning: a revalidation study of the reflection-in-learning scale. AB - The aims of this paper are to examine the measurement properties of the Reflection-in-Learning Scale (RLS) and to identify whether there are relationships between RLS scores early in the medical program and outcomes of the students' academic activity later on. The 14-item RLS was administered to second year students (N = 275) at start and at end of the third semester, after the students had reviewed their previous learning experience with the Course Valuing Inventory. The internal consistency, temporal stability and dimensionality of the RLS scores were investigated in relation to the start-end perspectives. Furthermore, a 2-year follow-up allowed the assessment of the relationships of third-term RLS scores with sixth-term measures of both academic achievement and diagnostic reasoning as appraised by the Diagnostic Thinking Inventory (DTI). Findings indicate that RLS data have acceptable unidimensionality and consistency of measurement, notwithstanding a significant individual-context interaction. Repeated measures revealed distinct patterns of RLS scores relating to perceived self-efficacy on the ability to reflect. Third-term RLS scores were significant, albeit weak, predictors of sixth-term cognitive achievement and DTI-related diagnostic reasoning ability. In conclusion, the results do not support a major explanatory role for RLS on knowledge representation. Nevertheless, the findings appear to substantiate the construct validity of this tool as an index of the students' frame of mind as regards reflective learning. They suggest that the RLS captures a self-regulation or cognitive housekeeping dimension of the students' reflective learning. The individual pattern of such (reflective) activity is likely to vary with specific learning conditions. PMID- 16362620 TI - Problem-based learning and learning approach: is there a relationship? AB - AIM: To assess the influence of a graduate-entry PBL curriculum on individual learning style; and to investigate the relationship between learning style, academic achievement and clinical reasoning skill. METHOD: Subjects were first year medical students completed the Study Process Questionnaire at the commencement, and again, at the end of the academic year when they also completed the Diagnostic Thinking Inventory, a measure of clinical reasoning skill. Subjects were classified on the basis of their predominant learning approach, and this was correlated with examination results and DTI score. RESULTS: There was a net shift in predominant learning approach away from deep learning towards a more surface approach over the period of the study, as well as a significant decrease in deep-learning scores. There was a statistically significant association between deep learning score and clinical reasoning skill as shown by total DTI score as well as on the structure of knowledge subscale. No correlation was found between learning approach and examination results. CONCLUSION: Although these results suggest that a deep learning approach may be beneficial in the development of clinical reasoning skill through its potential to enhance the development of knowledge representations, the substantial shift towards a surface learning approach brings into question previous conclusions that PBL curricula foster a deep approach to learning, and suggests that other factors, such as work load may be more determinants of learning approach than curriculum type. Taken together, these findings emphasise the context-dependent nature of learning approach as well as the importance of assessment as a driver of student learning and strongly suggest that further work to determine precisely the factors which influence learning approach in medical students is urgently needed. PMID- 16362621 TI - Correlates of reflective practice in medicine. AB - BACKGROUND: The ability of physicians to critically reflect on their professional practice has been increasingly valued. Previous research brought to light the multidimensional structure of reflective practice in medicine. It comprises at least five sets of behaviours in response to complex medical problems encountered in professional practice. Factors associated to reflective practice among physicians have, as far as we know, not yet been explored by empirical study. PURPOSE: To study factors correlated to reflective practice among physicians. METHODS: A questionnaire exploring characteristics of professional practice and educational experiences was administered to primary health care physicians. Measurements were related to scores on a reflective practice measuring instrument developed previously. Associations between variables were examined by statistical analysis with tests of correlation and analysis of variance. RESULTS: Reflective practice is negatively correlated to physician's age and number of years of clinical practice. Working mainly in hospitals and attendance to medical residency programmes in some specialties apparently have a positive effect on reflective practice. CONCLUSION: Reflective practice tends to decrease with experience. Findings are consistent with the literature on medical expertise that shows a decline of analytical reasoning in proportion to the increase in experience. Some specialty programmes seems to enhance concerns with the scientific basis to professional practice, thereby favouring reflective approaches. Local features of primary health care settings probably explain their negative effect on reflective practice. Strategies to develop reflective practice among physicians should be explored by further research. PMID- 16362622 TI - Emotional intelligence medical education: measuring the unmeasurable? AB - The construct of emotional intelligence (EI) has gained increasing popularity over the last 10 years and now has a relatively large academic and popular associated literature. EI is beginning to be discussed within the medical education literature, where, however, it is treated uncritically. This reflections paper aims to stimulate thought about EI and poses the question: Are we trying to measure the unmeasurable? The paper begins with an outline of the relevance and meaningfulness of the topic of EI for doctors. It continues with an overview of the main models and measures of EI. We then critique the psychometric properties of EI measures and give an illustrative case study where we tested the psychometric properties of the ECI-U with medical students. After our critique, we present an alternative model of EI and outline possible future directions for educational research. PMID- 16362626 TI - Neuromelanin induces oxidative stress in mitochondria through release of iron: mechanism behind the inhibition of 26S proteasome. AB - Parkinson's disease is characterized by the selective depletion of dopamine neurons in the substantia nigra, particular those containing neuromelanin. Involvement of neuromelanin in the pathogenesis may be either cytotoxic or protective. Recently we found that neuromelanin reduces the activity of 26S proteasome. In this paper, the detailed mechanisms behind the reduced activity were studied using neuromelanin isolated from the human brain. Neuromelanin increased the oxidative stress, but synthetic melanin did not. Superoxide dismutase and deferoxamine completely suppressed the increase, indicating that superoxide produced by an iron-mediated reaction plays a central role. Iron was shown to reduce in situ 26S proteasome activity in SH-SY5Y cells and the reduction was protected by antioxidants. These results suggest that iron released from neuromelanin increases oxidative stress in mitochondria, and then causes mitochondrial dysfunction and reduces proteasome function. The role of neuromelanin is discussed in relation to the selective vulnerability of dopamine neurons in Parkinson's disease. PMID- 16362627 TI - Influence of noradrenaline denervation on MPTP-induced deficits in mice. AB - C57/BL6 mice were administered either DSP4 (50 mg/kg, s.c., 30 min after injection of zimeldine, 20 mg/kg, s.c.) or vehicle (saline) at 63 days of age. Three weeks later, one group (n = 10) of DSP4-treated and one group of vehicle treated mice were administered MPTP (2 x 40 mg/kg, s.c., 24 hours between injections; the High dose groups), one group (n = 10) of DSP4-treated and one group of vehicle-treated mice were administered MPTP (2 x 20 mg/kg, s.c., 24 hours between injections; the Low dose groups), and one group (n = 10) of DSP4 treated and one group of vehicle-treated mice were administered vehicle. Three weeks later, all six groups were tested in motor activity test chambers, followed by injections of L-Dopa (20 mg/kg, s.c.), and then tested over a further 360 min in the activity test chambers. It was found that pretreatment with the selective NA neurotoxin, DSP4, deteriorated markedly the dose-dependent motor activity deficits observed in the vehicle pretreated MPTP treated mice. These 'ultra deficits' in the spontaneous motor behaviour of MPTP-treated mice were observed over all three parameters: locomotion, rearing and total activity, and were restricted to the 1(st) and 2(nd) 20-min periods. Administration of L-Dopa (20 mg/kg) following the 60-min testing of spontaneous behaviour restored the motor activity of Vehicle + MPTP treated mice (neither the Vehicle + MPTP-Low nor the Vehicle + MPTP-High groups differed from the Vehicle-Vehicle group, here) but failed to do so in the DSP4 pretreated mice. Here, a dose-dependent deficit of L Dopa-induced motor activity (over all three parameters) was obtained thereby offering further evidence of an 'ultra-deficit' of function due to previous denervation of the NA terminals. The present findings support the notion that severe damage to the locus coeruleus noradrenergic system, through systemic DSP4, disrupts the facilitatory influence on the nigrostriatal DA system, and interferes with the ability of the nigrostriatal pathway to compensate for or recover from marked injury, MPTP treatment. PMID- 16362628 TI - Influence of bilateral Stn-stimulation on psychiatric symptoms and psychosocial functioning in patients with Parkinson's disease. AB - Deep brain stimulation of the subthalamic nucleus is an effective treatment for advanced Parkinson's disease. There is some evidence that subthalamic stimulation not only affects motor function, but also mood, behaviour and cognition. In the present study we investigated the effects of subthalamic stimulation on psychiatric symptoms and psychosocial functioning in a consecutive series of patients with Parkinson's disease. 33 patients were assessed three times prior to surgery and at three, nine weeks as well as three, six and twelve months after surgery. We found significant improvements in depression, anxiety, psychological symptoms and distress after surgery. In most cases the amelioration followed surgery and was stable in the course of time. Individual analysis indicated deterioration in three patients despite motor improvement. The results suggest that stimulation of the subthalamic nucleus has a positive influence on psychiatric symptoms, psychosocial functioning and distress. We observed a decline in a minority of patients. PMID- 16362629 TI - Stage-dependent BDNF serum concentrations in Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by cognitive decline and loss of neurons in specific brain regions. Recent findings have suggested an involvement of brain-derived neurotrophic factor (BDNF) in the pathogenesis of AD. BDNF is an endogenous protein involved in the maintenance of neuronal function, synaptic plasticity and structural integrity in the adult brain. To our knowledge, the present pilot study assessed for the first time BDNF serum and CSF concentrations in 30 patients with different stages of AD in comparison to 10 age-matched non demendet controls. AD patients were divided in two groups according to their MMSE score: Group 1 (n = 15) in early stages with MMSE scores >or=21 (mean of 25.5) and Group 2 (n = 15) with more severe stages of dementia with MMSE scores <21 (mean of 13.3). As main results, we found in patients with early stages of probable AD significantly increased BDNF serum concentrations as compared to more severe stages of AD (p < 0.0001) and age-matched healthy controls (p = 0.028). BDNF serum values in all AD patients correlated significantly with MMSE scores (r = 0.486; p < 0.0001). Levels of BDNF were below the detection limit of the assay in unconcentrated CSF samples of AD patients and non-demendet controls.In summary, BDNF serum values are increased in early stages of Alzheimer's disease, which may reflect a compensatory repair mechanism in early neurodegeneration and could also contribute to increased degradation of beta-amyloid (Abeta). During the course of the disease, BDNF is decreasing, which correlates with the severity of dementia. The decrease of BDNF may constitute a lack of trophic support with an increase of Abeta accumulation and thus contribute to progressive degeneration of specific regions in the AD-affected brain. BDNF should be further evaluated as a candidate marker for clinical diagnosis and therapeutic monitoring in Alzheimer's disease. PMID- 16362630 TI - Neurochemical and oedematous changes in 1,3-dinitrobenzene-induced astroglial injury in rat brain from a 1H-nuclear magnetic resonance perspective. AB - 1,3-Dinitrobenzene (1,3-DNB), an intermediate used in the chemical industry, has toxic effects in the brain and testes. It produces focal lesions with marked astroglial necrosis in the rat brain upon repeated administration. Astrocytic death occurs in parallel with elevated local blood flow and is followed by damage to the cerebral vasculature and neurones. (1)H-nuclear magnetic resonance spectroscopic analysis before the onset of astrocytic damage, showed a global elevation of lactate, whereas choline containing compounds increased in the non vulnerable cerebral cortex, yet decreased in the vulnerable brainstem. Similarly, glutamate increased in the cerebral cortex, cerebellum and midbrain, but decreased in the susceptible brainstem. In vivo T2-weighted NMR imaging showed high signal intensities in brain nuclei shown to develop astroglial loss by conventional neuropathology at 24 hours after completion of dosing, but not at 6 10 hours. Hence the early neurochemical changes in susceptible areas contribute to the aetiology of degeneration, and those seen elsewhere may represent adaptive responses dependent on the particular phenotype of different cell groups and underlying metabolic relationships. PMID- 16362631 TI - Association of ghrelin receptor gene polymorphism with bulimia nervosa in a Japanese population. AB - Eating disorders (EDs) have a highly heterogeneous etiology and multiple genetic factors might contribute to their pathogenesis. Ghrelin, a novel growth hormone releasing peptide, enhances appetite and increases food intake, and human ghrelin plasma levels are inversely correlated with body mass index. In the present study, we examined the 171T/C polymorphism of the ghrelin receptor (growth hormone secretagogue receptor, GHSR) gene in patients diagnosed with EDs, because the subjects having ghrelin gene polymorphism (Leu72Met) was not detected in a Japanese population, previously. In addition, beta3 adrenergic receptor gene polymorphism (Try64Arg) and cholecystokinin (CCK)-A receptor (R) gene polymorphism (-81A/G, -128G/T), which are both associated with obesity, were investigated. The subjects consisted of 228 Japanese patients with EDs [96 anorexia nervosa (AN), 116 bulimia nervosa (BN) and 16 not otherwise specified (NOS)]. The age- and gender-matched control group consisted of 284 unrelated Japanese subjects. The frequency of the CC type of the GHSR gene was significantly higher in BN subjects than in control subjects (chi(2) = 4.47, p = 0.035, odds ratio = 2.05, Bonferroni correction: p = 0.070), while the frequency in AN subjects was not different from that in controls. The distribution of neither beta3 adrenergic receptor gene nor CCK-AR polymorphism differed between EDs and control subjects. Therefore, the CC type of GHSR gene polymorphism (171T/C) is a risk factor for BN, but not for AN. PMID- 16362632 TI - Withdrawal from repeated amphetamine administration leads to disruption of prepulse inhibition but not to disruption of latent inhibition. AB - The present study represents a continuous effort to develop an animal model of schizophrenia based on the "endogenous dopamine sensitization" hypothesis. To achieve this goal, withdrawal from an escalating amphetamine (AMPH) regime administration [three injections per day over a period of 4 days and increasing doses from 1 to 10 mg/kg of AMPH or an equivalent volume of saline (SAL)] was employed. Animals exposed to this treatment were evaluated on their performance in attentional (Latent inhibition, LI) and sensorimotor gating (Prepulse inhibition, PPI) tasks in a drug free state and tested for locomotor sensitization following a low dose of AMPH challenge administration.LI using active avoidance, tested on withdrawal day 4, was unaffected. PPI of the acoustic startle response, measured on withdrawal days 6 and 70, was disrupted. On the 76th day of withdrawal, a low challenge dose of AMPH (1 mg/kg) led to a clear locomotor sensitization effect. PMID- 16362633 TI - Association study of polymorphisms in LRP1, tau and 5-HTT genes and Alzheimer's disease in a sample of Colombian patients. AB - Analysis of genetic susceptibility factors for Alzheimer's disease (AD) in populations with different genetic and environmental background may be useful to understand AD etiology. There are few genetic association studies of AD in Latin America. In the present work, we analyzed polymorphisms in 3 candidate genes; the LDL receptor related protein-1, the microtubule-associated protein Tau and the serotonin transporter genes in a sample of 106 Colombian AD patients and 97 control subjects. We did not find a significant allelic or genotypic association with any of the three polymorphisms analyzed using different statistical analysis, including a neural network model or different sample stratifications. To date, APOE polymorphisms are the only genetic risk factors identified for AD in the Colombian population. It may be factible that future combination of high throughput genotyping platforms and multivariate analysis models may lead to the identification of other genetic susceptibility factors for AD in the Colombian population. PMID- 16362634 TI - Neuropathological changes in ten cases of neuronal intermediate filament inclusion disease (NIFID): a study using alpha-internexin immunohistochemistry and principal components analysis (PCA). AB - Ten cases of neuronal intermediate filament inclusion disease (NIFID) were studied quantitatively. The alpha-internexin positive neurofilament inclusions (NI) were most abundant in the motor cortex and CA sectors of the hippocampus. The densities of the NI and the swollen achromatic neurons (SN) were similar in laminae II/III and V/VI but glial cell density was greater in V/VI. The density of the NI was positively correlated with the SN and the glial cells. Principal components analysis (PCA) suggested that PC1 was associated with variation in neuronal loss in the frontal/temporal lobes and PC2 with neuronal loss in the frontal lobe and NI density in the parahippocampal gyrus. The data suggest: 1) frontal and temporal lobe degeneration in NIFID is associated with the widespread formation of NI and SN, 2) NI and SN affect cortical laminae II/III and V/VI, 3) the NI and SN affect closely related neuronal populations, and 4) variations in neuronal loss and in the density of NI were the most important sources of pathological heterogeneity. PMID- 16362635 TI - Very long term studies of the seeding of beta-amyloidosis in primates. AB - Cerebral beta-amyloidosis was found in 16/18 marmosets aged <10 yrs and 8/9 marmosets aged >10 yrs, injected intracerebrally with human or marmoset brain homogenate containing beta-amyloid 1-8 years previously. It was found in only 2/12 marmosets aged <10 yrs and 1/15 marmosets aged >10 yrs, injected with synthetic Abeta-peptides, CSF, or brain tissue which did not contain beta amyloid. Cerebral beta-amyloidosis was found in 0/11 uninjected marmosets aged <10 yrs and in 5/29 uninjected marmosets aged >10 yrs. The beta-amyloidosis comprised small and large vessel angiopathy and some plaques throughout cortex and was qualitatively similar in injected marmosets and, when present, in uninjected marmosets. Of those injected marmosets which were positive, the amount of beta-amyloidosis was unrelated to age or incubation times but the 3 injected marmosets without beta-amyloidosis had incubation times of <3.5 years. We conclude that beta-amyloid, or associated factors, can initiate or accelerate the process of cerebral amyloidosis in primates. PMID- 16362637 TI - The effect of psychostimulants on [3H]dopamine uptake and release in rat brain synaptic vesicles. AB - Amphetamine and its derivatives are psychostimulants active at the plasma membrane monoamine transporters. In the present study we assessed the interaction of parachloroamphetamine, D-amphetamine, fenfluramine and methylendioxymethamphetamine with brain vesicular monoamine transporter using purified rat striatal synaptic vesicles. All four psychostimulants inhibited vesicular [(3)H]dopamine uptake in a competitive and dose-dependent manner and had no effect on [(3)H]dihydrotetrabenazine binding. At higher concentrations the drugs enhanced [(3)H]dopamine vesicular efflux. Parachloroamphetamine was the most potent agent while methylendioxymethamphetamine was the weakest one. The vesicular activities may be relevant to their neurotoxicity. PMID- 16362636 TI - Neuroprotection of cerebrolysin in tissue culture models of brain ischemia: post lesion application indicates a wide therapeutic window. AB - All attempts to reduce neuronal damage after acute brain ischemia by the use of neuroprotective compounds have failed to prove efficacy in clinical trials so far. One of the main reasons might be the relatively narrow time window for intervention. In this study 2 different tissue culture models of ischemia, excitotoxic lesion by the use of glutamate and oxygen-glucose deprivation (OGD), were used to investigate the effects of delayed application of Cerebrolysin (Cere) on neuronal survival. This drug consists of low molecular weight peptides with neuroprotective and neurotrophic properties similar to naturally occurring growth factors. After both types of lesion, acute as well as delayed treatment with Cere resulted in a dose dependent and significant rescue of neurons. In the model of excitotoxic cell death significant drug effects were found even when the treatment started with a delay of 96 hours after addition of glutamate. In the OGD model pronounced effects were found after 48 hours delay of treatment, and even after 72 hours a small but significant rescue of neurons was detected. The neuroprotective effects of a single addition of Cerebrolysin to the culture medium resulted in significant protection until end of the experiments which was up to 2 weeks after the initial lesion. A shift of the efficacious dosages from low to high concentrations indicates that most likely active compounds are used up, indicating that multiple dosing might even increase the effect size. In conclusion the results indicate that Cere displays a relatively wide therapeutic time window which might be explained by a combination of acute neuroprotective properties and neurotrophic efficacy. PMID- 16362639 TI - Inferring candidate genes for attention deficit hyperactivity disorder (ADHD) assessed by the World Health Organization Adult ADHD Self-Report Scale (ASRS). AB - The present study tests the psychometric properties and validity of the German version of the World Health Organization Adult Attention Deficit Hyperactivity Disorder (ADHD) Self-Report Scale (ASRS), which is a short screening instrument for use in the general population. Furthermore, two candidate genes for ADHD, the COMT VAL158MET and the 5-HT2a T102C polymorphisms, were tested for associations with the ASRS subscales inattention and hyperactivity/impulsivity in N = 203 healthy subjects. The ordinal CFA yielded a two-factorial model corroborating the structure of the official English WHO version. Genetic analysis revealed an association between the VAL allele of COMT and the inattention scale (F(1, 201) = 7.20, p = 0.008), the hyperactivity/impulsivity scale (F(1, 201) = 4.30, p = 0.039), and the total ASRS scale (F(2, 201) = 7.64, p = 0.006) with highest scores in carriers of the MET/MET genotype. The C-allele of 5-HT2a was significantly associated with the hyperactivity/impulsivity scale (F(1, 201) = 5.52, p = 0.020) and the total ASRS scale (F(1, 201) = 4.21, p = 0.042) with highest scores in carriers of the TT genotype. The data provide evidence for the structural as well as for the external validity of the ASRS. PMID- 16362640 TI - A movement protein and three capsid proteins are all necessary for the cell-to cell movement of apple latent spherical cheravirus. AB - Immunoblot analysis of apple latent spherical cheravirus (ALSV)-infected leaves using a polyclonal antibody against the 21 C-terminal amino acids of a 53 K/42 K movement protein (MP) showed that a protein with an Mr of 42 kDa (42KP) is the dominant form found in vivo, which could indicate that the second AUG is used as an initiation codon of a ORF in RNA2. Co-expression of GFP with 42KP in tobacco epidermal cells showed that 42KP is able to facilitate cell-to-cell trafficking of GFP that is expressed in the same cells. The analysis of deletion mutants on each of MP, Vp24, Vp20, or Vp25 using an ALSV vector that stably expresses GFP indicated that an MP and three capsid proteins are all indispensable for the cell to-cell movement of the virus. In ultrathin sections of infected leaves, a file of virus-like particles passing through the plasmodesmata connecting neighboring cells and tubular structures containing virus-like particles extending into the cytoplasm were observed. These results show that ALSV moves from cell to cell as virus particles. PMID- 16362638 TI - Regional acetylcholinesterase activity and its correlation with behavioral performances in 15-month old transgenic mice expressing the human C99 fragment of APP. AB - In addition to Abeta plaques and neurofibrillary tangles, Alzheimer's disease (AD) is characterized by increased brain levels of APP C-terminal fragments. In the present investigation, the cholinergic innervation in forebrain regions of transgenic mice (Tg13592) expressing the human betaAPP C99 fragment was compared to that of non-transgenic controls by measuring the activity of the non-specific catabolic enzyme, acetylcholinesterase (AChE). The AchE activity of Tg13592 mice was altered in several regions implicated in the functional loop of regulation between septum and hippocampus, vulnerable in Alzheimer pathology and critically involved in cognitive functions. In particular, AChE activity was upregulated in three basal forebrain regions containing cholinergic cell bodies, prelimbic cortex, anterior subiculum, and paraventricular thalamus, but downregulated in lateral septum and reticular thalamus. The increased activity in medial septum and anterior subiculum was linearly correlated with poor performances in a spatial learning task, possibly due to cell stress mechanisms. Because of some similarities in terms of neurochemistry and behavior, this mouse model may be of use for studying prodromal AD. PMID- 16362641 TI - Genetic stability within the Norwegian subtype of salmonid alphavirus (family Togaviridae). AB - Salmonid alphavirus (SAV) (family Togaviridae) causes mortality in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss W.) in Norway, France, UK, and Ireland. At least three subtypes of SAV exist: SPDV in UK/Ireland, SDV in France/UK, and the recently reported Norwegian salmonid alphavirus (NSAV) in western Norway. During 2003 and 2004, disease caused by NSAV was reported for the first time in northern Norway, more than 800 km away from the enzootic area in western Norway. The present study has investigated the phylogenetic relationships among 20 NSAV isolates, based on a 1221-nt-long segment covering part of the capsid gene, E3, and part of the E2 gene, collected over a period of eight years. The results revealed genetic homogeneity among NSAV isolates, including those from northern Norway. The SDV or SPDV subtypes were not found in diseased Norwegian fish. A substitution rate of 1.70 (+/-1.03) x 10(-4) nt subst/site/year was obtained for the NSAV subtype by maximum likelihood analysis. The second aim of this study was to clarify whether NSAV changes genotypically in cell culture by culturing a NSAV isolate through 20 passages in CHSE-214 cells. Sequencing of almost the entire genome (11530 nt) after 20 passages revealed four nucleotide substitutions, all resulting in amino acid substitutions. One of these substitutions, serine to proline in E2 position 206, was also found to have occurred in field isolates. PMID- 16362642 TI - High incidence of elevated antibody titers to Epstein-Barr virus in patients with uveitis. AB - We assayed Epstein-Barr virus (EBV) antibody titers in patients' sera using indirect immunofluorescence and tested for the presence of antibody to EBV immediate-early BZLF1 protein ZEBRA by Western blotting to explore the association of EBV infection with uveitis. IgG and IgA antibodies to viral capsid antigen (VCA), IgG antibodies to early antigen (EA), and antibodies to EBV nuclear antigen were detected at higher titers in sera of patients with uveitis than in the sera of healthy controls. Neither IgM antibody to VCA nor EA was detected in the patients' sera. Anti-ZEBRA-IgG antibodies were detected in most patients' sera, but not in those of healthy controls. These results suggest that uveitis might be a disease accompanied by EBV reactivation. PMID- 16362643 TI - Viral reactivation and pseudotype production in an in vitro superinfection system with two different strains of HIV-1. AB - Viral production and variability of HIV-1 is normally high in vivo causing the necessary conditions for cellular superinfection. In order to evaluate the superinfection dynamics in vitro, H9HTLVIIIB cell line was superinfected with HIVMN. Superinfected cells showed nearly 50% cell mortality at day 1 post superinfection (ps), which increased significantly up to day 4 ps. Superinfecting genome was detectable until day 10 ps. The superinfecting strain was found in the supernatant only on day 1 ps, but was recovered up to day 4 ps by coculture with non-infected cells. The existing strain (HIVHXB2) was recovered throughout the studied period. Pseudotype formation by the HIVHXB2 genome and envelope proteins of the superinfecting strain (HIVMN) was observed from day 1 to 6 ps. Viral production was increased by 1.7 LOG in superinfected cells from day 1 ps. Both viral production increase and pseudotype formation could be relevant for HIV pathogenesis in vivo. PMID- 16362644 TI - CGRP and migraine: neurogenic inflammation revisited. AB - For more than a century neurogenic inflammation has been proposed to have a role in various human diseases. The present review will cover the conceptual steps of the itinerary that has led to the conclusion that neurogenic inflammation is important in migraine. Of particular relevance for the object of this article is the observation that tachykin-independent neurogenic inflammatory responses are evident in rodents, but much less pronounced or absent in other mammal species, including man, whereas neurogenic vasodilatation, most likely mediated by CGRP, occurs in most mammalian species and also in man. Recent evidence that a CGRP receptor antagonist was effective in the treatment of migraine attack supports the hypothesis that neurogenic vasodilatation is a major underlying mechanism of migraine. PMID- 16362645 TI - High prevalence of patent foramen ovale in migraine with aura. AB - In this study we evaluated the presence of patent foramen ovale (PFO) in a cohort of 25 consecutive patients suffering from migraine with aura (MA) during an attack presenting to the emergency ward of an Italian hospital. Patients underwent brain magnetic resonance imaging (MRI) with contrast medium, routine coagulation tests, contrast transcranial echocolour-coded sonography (c-TCCS) and transoesophageal echocardiography (TEE). Of the enrolled patients, 88.7% showed a PFO according to the c-TCCS test, whereas only in 72% TEE confirmed the presence of PFO. This discordance could be due to the fact that c-TCCS is more sensitive even with shunts with minimal capacity also located in the pulmonary vasculature. After surgical treatment of the PFO, MA disappeared within two months. Also, the treatment with warfarin as well as with acetylsalicylic acid and flunarizine was able to dramatically reduce the frequency of migraine attacks. These data indicate a higher prevalence of PFO in MA vs. normal population (OR=2.92) and could suggest that the presence of arteriovenous (AV) shunts could represent a trigger for MA attacks as well as for stroke, but more studies are needed to confirm this preliminary hypothesis. PMID- 16362646 TI - Topiramate in the treatment of refractory chronic daily headache. An open trial. AB - Chronic daily headache (CDH) is a debilitating disorder that becomes a treatment challenge in patients refractory to the treatment. We hereby report our experience with topiramate treatment in patients with refractory CDH. The study design was a prospective, protocol-based follow-up and retrospective analysis of headache diaries. We treated with topiramate at slowly increased moderate increments 11 CDH patients who were refractory to multiple previous treatments. Topiramate treatment was effective in 7 (64%) patients. The treatment resulted in a 66% (median) decrease of the headache days per week and a significant decrease in headache severity, a reduction of the headache hours per day, and weekly analgesic consumption. These effects continued for an average follow-up of 8+/-4 months. The average effective dose was 100 mg/day. Slowly increasing the drug at moderate increments resulted in high tolerability of topiramate. We found topiramate to be an effective long-standing treatment option for patients with refractory CDH. Slow increments of the dosage contributed to high tolerability of the drug. PMID- 16362647 TI - Managing migraine headaches experienced by patients who self-report with menstrually related migraine: a prospective, placebo-controlled study with oral sumatriptan. AB - The objective was to evaluate the efficacy and tolerability of oral sumatriptan (100 mg) in patients who self-reported with menstrually related migraine. A prospective, multicentre, randomised, double-blind, placebo-controlled, two-group crossover study was carried out in 20 UK primary and secondary care surgeries. Of 115 patients with a self-reported history of menstrually related migraine that entered the study, 93 patients completed it. Patients treated all migraine attacks for 2 months with sumatriptan (100 mg) and for 2 months with placebo. The primary endpoint was the proportion of patients reporting headache relief at 4 hours for the first treated attack. Only 11% of patients fulfilled the protocol definition of menstrually related migraine. Patients reported a variable pattern of migraine attacks occurring inside and outside the menstrual window. For the first attack, significantly more patients receiving sumatriptan than placebo reported headache relief for attacks occurring inside (67% vs. 33%, p=0.007) and outside (79% vs. 31%, p<0.001) the menstrual period. Sumatriptan was generally well tolerated. Oral sumatriptan (100 mg) is an effective and well tolerated acute treatment for patients who report menstrually related migraine. PMID- 16362648 TI - Cluster-like headache after surgical crystalline removal and intraocular lens implant: a case report. AB - Cluster headache (CH) is a well characterized primary headache disorder. Nevertheless, symptomatic CH has been reported in association with various underlying diseases. Symptomatic cluster headache related to ocular pathologies have been rarely described. We report a case consequent to a surgical operation for cataract. PMID- 16362649 TI - A woman with daily headaches. AB - Headache, and migraine in particular, is the main neurological reason for consultation. We present the case of a 48-year-old woman who experienced a transformation of her episodic migraine attacks into daily headache episodes due to the ingestion of biscuits containing wheat as their main ingredient. This experience emphasises that a good clinical interview remains the most important point in the diagnosis and management of headache. PMID- 16362650 TI - Migraine with aura, bipolar depression, ACM aneurysm. A case report. AB - B.D. is a 48-year-old professional woman. She has been suffering for migraine since she was 28, but she did not have serious problems until last year, when headache episodes became more frequent and it was necessary an admission to emergency room. At the beginning, the events were about 6 per month, lasting from 2 to 4 days, beating and of high intensity together with nausea, vomit, photo and phonophobia and visual area. Looking at the anamnesis, we report a psychiatric treatment since about ten years, because of type II bipolar disorder. In spite of the psychopharmacological treatment, as the patient came in our Regional Headache Center, she talked about 7 events with aura (scintillating scotomas, emianopsia) per month, lasting 2-4 days with photo and phonophobia, nausea, crying crisis, anxiety. Although the neurological examination was normal, the sudden aggravation of pain symptomatology and the unresposiveness to usual painkillers, suggested a cerebral CT and CT-angiography. CT and CT-angiography discovered the presence of an aneurysm of the right middle cerebral artery (MCA) of 4 mm diameter, with parietal irregularities. The patient was operated to reduce the hemorrhagic risk, with a positive result. One year after the operation, the patient reports a decrease of headache events with a frequency of 2 per month, lasting only a few hours, which she can now solve with COXIB. PMID- 16362651 TI - Acupuncture use for the treatment of headache prior to neurological referral. AB - Acupuncture is a popular complementary treatment for various pain syndromes. Some studies claim efficacy in the treatment of primary headache syndromes. However, data on the frequency of acupuncture use by patients with headache prior to neurological referral have not been identified. In this study, 12% of patients with headache attending general neurology outpatient clinics had already received acupuncture; of the remainder, 73% said they would be willing to try it. PMID- 16362652 TI - Conversion from migraine without aura to typical aura without headache after irbesartan. PMID- 16362654 TI - Padua, the cradle of modern medicine: Bernardino Ramazzini (1633-1714) on headaches. AB - A prominent historian of Medicine, Henry Sigerist, quoted the Padua Medical School as "the cradle of modern medicine". This opinion is currently accepted worldwide. A short outline on the contribution of the Padua Medical School to the development of medical knowledge in its "golden age" is given. In this context, the work of a prominent figure of the 17(th) century Padua University and the founder of Occupational Medicine, Bernardino Ramazzini, is considered, with focus on his interest in headache. PMID- 16362655 TI - Headaches in pregnancy. AB - Most women with migraine improve during pregnancy. Some women have their first attack. Migraine often recurs postpartum and can begin for the first time. Drugs are commonly used during pregnancy despite insufficient knowledge about their effects on the growing fetus. Most drugs are not teratogenic. Adverse effects, such as spontaneous abortion, developmental defects and various postnatal effects depend on the dose and route of administration and the timing of the exposure relative to the period of foetal development. While medication use should be limited, it is not absolutely contraindicated in pregnancy. Nonpharmacologic treatment is the ideal solution; however, analgesics such as acetaminophen and opioids can be used on a limited basis. Preventive therapy is a last resort. PMID- 16362656 TI - Chronic headaches: from research to clinical practice. AB - Chronic daily headache (CDH) is a heterogeneous group of headaches that includes primary and secondary varieties. Primary CDH is a frequent entity that probably affects 4-5% of the population. It can be subdivided into headaches of short duration (<4 h/attack) like chronic cluster headache, and disorders of long duration (>4 h/attack). Primary CDH of long duration includes transformed migraine, chronic tension-type headache, and new daily persistent headache and hemicrania continua. Analgesics, ergots and triptan overuse are frequent in all types of CDH. We revise recent insights into the epidemiology, pathophysiology, clinical characteristics and prognosis of CDH. PMID- 16362657 TI - Genetic risk factors in primary paediatric versus adult headache: complexities and problematics. AB - Numerous candidate genes for migraine have been proposed on the basis of their possible functional role in its pathogenesis. Genetic polymorphisms have been evaluated in association studies, some of which have been suggested to be susceptibility markers for adult migraine. To date, however, none of the identified polymorphisms in adult migraine susceptibility have been investigated in children, raising the possibility that they may not be necessarily involved in paediatric migraine susceptibility. This paper reviews studies of the genetic basis of migraine and summarises our experience in genetic association studies in primary paediatric headache susceptibility. PMID- 16362658 TI - Functional serotonin 5-HTTLPR polymorphism is a risk factor for migraine with aura. AB - In the present work, we report that the functional serotonin transporter gene promoter (5-HTTLPR) polymorphism is involved in migraine pathogenesis. The distribution of 5-HTTLPR genotypes was significantly different in MA patients (S/S vs. S/L vs. L/L=32.7 vs. 42.3 vs. 25.0%), MO patients (18.5 vs. 39.1 vs. 42.4%) and CON (18.0 vs. 51.3 vs. 30.7%; chi-square test, p<0.05). In 5-HTTLPR S/S carriers, the odds ratio for MA risk was 2.60 (95% confidence interval [95%CI]=1.75-3.85) compared to CON, and it was 2.14 (95%CI=1.42-3.21) compared to MO. These data provide a further insight on the complex genotype-phenotype relationship involved in MA pathogenesis, and might eventually result in new and individualised prognostic and therapeutic measures. PMID- 16362659 TI - Association between migraine and HLA-DRB1 gene polymorphisms. AB - We examined the distribution of HLA-DRB1 alleles in a cohort of 255 Italian migraine patients and in a control group of 325 healthy subjects. The frequency of DRB1*12 allele was found to be significantly reduced (p=0.02) in patients with migraine while the DRB1*16 allele was significantly increased (p=0.04) in comparison with controls. When the patients were divided into disease subgroups (migraine with and without aura), HLA-DRB1**16 allele was significantly increased (p<0.05) only in migraine without aura patients. We conclude that, in Italian patients, migraine is associated with different alleles of the HLA-DRB1 locus. Our data suggest the presence of a genetic susceptibility factor for migraine within the HLA region. PMID- 16362660 TI - Cytotoxic T lymphocyte antigen 4 polymorphism 49 (A>G) and migraine. AB - Migraine without aura (MO) and migraine with aura (MA) are disorders involving multiple environmental and genetic factors. The A/G polymorphism located within exon 1 of the gene encoding the cytotoxic T lymphocyte antigen 4 (CTLA-4) is associated with several HLA-associated multifactorial diseases. The CTLA-4 family shows a negative control on T-cell proliferation and cytokine production (TNF alpha and IL-10). In the present study we investigated the contribution of the candidate gene CTLA-4 in migraine pathophysiology. Included in the study were 96 MO and 39 MA migraine patients and 106 healthy individuals as control group. The results showed no statistical difference of allele frequencies between patient group and control group. These results would indicate no association between MA and MO migraine and CTLA-4 polymorphism, excluding any possible role of the CTLA 4 gene as a genetic factor determining susceptibility to migraine. PMID- 16362661 TI - Neurophysiological approach to central pain modulation in primary headaches. AB - The study of CNS pain-modulating pathways has led to important discoveries about the role of central nociceptive structures such as PAG and hypothalamus in the pathophysiology of episodic and chronic primary headaches. Functional neuroimaging studies have revealed that primary headaches are characterised by different patterns of activation of central pain modulatory structures. A future model of headache pathophysiology investigating the contribution of CNS pain modulating pathways will probably increase our understanding of pain processing in primary headaches. Herein we review the neurophysiological approaches to assess central pain modulation in primary headaches with emphasis on the diffuse noxious inhibitory control, a form of endogenous pain inhibition. In addition, patients' data will be presented that highlights the utility of such methods for primary headache's pathophysiology and clinical monitoring. PMID- 16362662 TI - Habituation of single CO2 laser-evoked responses during interictal phase of migraine. AB - A reduced habituation of averaged laser-evoked potential (LEP) amplitudes was previously found in migraine patients. The aim of the present study was to assess the habituation of single LEP responses and pain sensation during the interictal phase in migraine patients. Fourteen migraine patients were compared with ten control subjects. The pain stimulus was laser pulses, generated by CO2 laser, delivered to right supraorbital zone. Patients were evaluated during attack-free conditions. The LEP habituation was studied by measuring the changes of LEP amplitudes across and within three consecutive repetitions of 21 non-averaged trials. In migraine patients the N2-P2 wave amplitudes did not show a tendency toward habituation across and, above all, within the three repetitions. Anomalous behaviour of nociceptive cortex during the interictal phase of migraine may predispose patients to headache occurrence and persistence. PMID- 16362663 TI - Medication-overuse headache: pathophysiological insights. AB - Medication overuse headache (MOH) is a clinically important entity and it is now well documented that the regular use of acute symptomatic medication by people with migraine or tensiontype headache increases the risk of aggravation of the primary headache. MOH is one of the most common causes of chronic migraine-like syndrome. Because of easy availability and low expense, the greatest problem appears to be associated with barbiturate-containing combination analgesics and over-the-counter caffeine-containing combination analgesics. Even though triptan overuse headache is not encountered with great frequency, all triptans should be considered potential inducers of MOH. There are several different theories regarding the aetiology of MOH, including: (i) central sensitisation from repetitive activation of nociceptive pathways; (ii) a direct effect of the medication on the capacity of the brain to inhibit pain; (iii) a decrease in blood serotonin due to repetitive medication administration with alteration of serotonin receptors; (iv) cellular adaptation in the brain; and (v) changes in the periaqueductal grey matter. The principal approach to management of MOH is built around cessation of overused medication. Without discontinuation of the offending medication, improvement is almost impossible to attain. Thus, the best management advice is to raise awareness and strive for prevention. In this article, we analyse also the possible mechanisms that underlie sensitisation in MOH by comparing these mechanisms with those reported for other forms of drug addiction. PMID- 16362664 TI - Introduction to the new International Classification of Headache Disorders. AB - Based on the first edition of the classification (already validated), the ICHD-II is intended for not only the researcher and clinician but also the neurologist and general practitioner. All the headaches are classified into groups and subgroups to provide the level of diagnosis necessary for each user. PMID- 16362665 TI - Application of ICHD 2nd edition criteria for primary headaches with the aid of a computerised, structured medical record for the specialist. AB - We tested the computerised, structured medical record by entering and analysing the consecutive clinical sheets of primary headaches in the episodic forms (200) and chronic headache (200) and the corresponding output diagnoses of patients attending our Headache Centre. A diagnosis of one of the primary headache forms was obtained in 67.9% of cases. A certain diagnosis of primary headache plus that of a probable form was obtained in 24.4% of cases (12.7% represented by chronic migraine (CM) or chronic tension-type headache (CTTH)+probable medication overuse headache). Only probable forms were diagnosed in the remaining 7.3% (as single probable diagnosis in 5.8% of cases or multiple diagnoses of probable forms in the remaining ones). The percentage of certain diagnoses mainly in the chronic headache group (28.4%), and to a lesser extent tension-type headache (6.5%), were obtained in 34.9% of cases. A certain diagnosis of one chronic form plus that of a probable form was obtained in 50.8% of cases (26.9% represented by probable medication-overuse headache). Only probable forms were diagnosed in 13.46% (as single probable diagnosis in 8.73% of cases or multiple diagnoses of probable forms in the remaining ones). In the other cases, the ICHD-II classification does not allow the diagnoses of CM, CTTH or probable forms and medication overuse headache because the mandatory criteria for the diagnoses are too stringent and do not reflect modifications of the headache pattern in relation to its chronicity. These preliminary results underscore the usefulness of a computerised device based on the ICHD 2nd edition for diagnostic purposes in tertiary centres dedicated to headaches in clinical practice as well as its relevance for research. This computerised device may help to validate the new diagnostic criteria and to answer some emerging questions from the application of the new classification version, the relevance of which should be verified in clinical practice. PMID- 16362666 TI - Development of a ICHD-II based computerised system for the general practitioner. AB - We present a computerised programme designed for use in the office of a general practitioner. The system provides an assisted diagnosis according to the ICHD-II criteria for the principal forms of primary headaches (migraine, tension-type headache, cluster headache) and highlights the red flags of a possible secondary headache. A relevant feature is that explanations for the selection of a particular diagnosis are given at the end of the process; furthermore, the characteristics of the patient's headache, which were previously inserted in the programme by the physician, are summarised, allowing critical evaluation of the suggested diagnosis. The software can also be used as a clinical file, in that it is possible to create for each patient a clinical chart in which to record the selected diagnosis, the recommended therapy and any eventual comments. Our programme aims for educational growth, promoting the learning of the basic ICHD II criteria. PMID- 16362667 TI - Osmophobia in primary headaches. AB - This study evaluates osmophobia (defined as an unpleasant perception, during a headache attack, of odours that are non-aversive or even pleasurable outside the attacks) in connection with the diagnosis of primary headaches. We recruited 775 patients from our Headache Centre (566 females, 209 males; age 38+/-12 years), of whom 477 were migraineurs without aura (MO), 92 with aura (MA), 135 had episodic tension-type headache (ETTH), 44 episodic cluster headache (ECH), 2 chronic paroxysmal hemicrania (CPH) and 25 other primary headaches (OPHs: 12 primary stabbing headaches, 2 primary cough headaches, 3 primary exertional headaches, 2 primary headaches associated with sexual activity, 3 hypnic headaches, 2 primary thunderclap headaches and 1 hemicrania continua). Among them, 43% with MO (205/477), 39% with MA (36/92), and 7% with CH (3/44) reported osmophobia during the attacks; none of the 135 ETTH and 25 OPH patients suffered this symptom. We conclude that osmophobia is a very specific marker to discriminate adequately between migraine (MO and MA) and ETTH; moreover, from this limited series it seems to be a good discriminant also for OPHs, and for CH patients not sharing neurovegetative symptoms with migraine. Therefore, osmophobia should be considered a good candidate as a new criterion for the diagnosis of migraine. PMID- 16362668 TI - A validation study of an Italian version of the ID Migraine: preliminary results. AB - Migraine is a highly prevalent and disabling disease that is substantially undiagnosed in primary care. Recently, the ID Migraine, a self-administered questionnaire, was shown to be a valid and reliable screener for migraine in primary care in the USA. To validate an Italian version of the ID Migraine, we planned a multicentric study, evaluating at least 220 patients affected by various form of headache. The responses to the questionnaire were compared with the diagnosis of headache made by a headache specialist blind to the result of the questionnaire. Sensitivity, specificity, and positive and negative predictive values for migraine were calculated. The statistical analysis on 140 patients now examined showed a very good performance of the ID Migraine with high sensitivity: 0.94 (95% CI: 0.89-0.95), specificity: 0.70 (95% CI: 0.54-0.86) and positive predictive value: 0.89 (0.82-0.95). If confirmed, these results would establish ID Migraine as a valid screening instrument for migraine in Italian headache patients and warrant further investigation in primary care to assess the validity of this ID screener in Italian population. PMID- 16362669 TI - The comorbidity of migraine and hypertension. A study in a tertiary care headache centre. AB - Literature data concerning the comorbidity of migraine and hypertension are conflicting and lacking in consistency. This study was designed to evaluate the distribution of hypertension in a sample of migraineurs in comparison with a group of tension type headache (TTH) patients. Hypertension prevalence was more elevated in headache sufferers than in the general population. This finding might be due to a Berkson's bias; in fact individuals seeking medical care often show a high rate of association between two medical conditions which may be independent in the general population. The highest hypertension prevalence was found in patients with TTH, supporting the hypothesis that this type of headache might be based upon vascular mechanisms. PMID- 16362670 TI - Patients with headache and functional dyspepsia present meal-induced hypersensitivity of the stomach. AB - Headache is a frequent feature of functional gastrointestinal disorders but there is no data on the responsible pathophysiological mechanism. The aim of this study was to verify whether alteration of post-prandial gastric tone or sensitivity might explain this association. Fourteen patients affected by functional dyspepsia (7 migraine without aura) and 7 healthy volunteers (HV) underwent gastric tone measurement in fasting condition and after the administration of a liquid meal by barostat. Gastric volume (GV) and accommodation were calculated as difference between mean post-prandial and mean fasting volume. Mean postprandial GV increase and fasting perception and discomfort threshold (DTh) were similar among the 3 groups. DTh after meal was lower in dyspeptic headache patients than in HV and dyspeptic without headache patients. Patients with migraine and functional dyspepsia may be characterised by meal-induced hypersensitivity of the stomach. PMID- 16362671 TI - Psychiatric disorders and pain location in unilateral migraineurs. AB - To explore the relationship between the side of pain during attacks and psychopathological features in strictly unilateral migraine, we recruited 35 patients affected by migraine with and/or without aura diagnosed according to the revised ICHD-II criteria. Seventeen patients had right side-locked pain (R-SUM), 11 had left side-locked (L-SUM) and 7 had side-shifting pain (SSM). Patients were administered the Hamilton Anxiety Scale, the State and Trait Anxiety Inventory State Anxiety, the Beck Depression Inventory and the 20-item Toronto Alexithymia Scale. Statistical analyses showed that the L-SUM group was significantly more anxious and depressed than the other two groups of patients. Our preliminary data suggest that strictly left unilateral migraine is associated with more severe anxiety and depression. This finding appears to be consistent with studies that reported a higher degree of these disorders in patients with left cerebral hemispheric damage. PMID- 16362672 TI - Tertiary treatment for psychiatric comorbidity in headache patients. AB - The presence of significant and confounding psychiatric comorbidity is greater in patients attending headache clinics than in headache patients from the general population. The frequent comorbidity of headache with generalized anxiety disorder can take advantage of the administration of benzodiazepines. With regard to depression-related headache, it's wellknown that the antidepressive drugs can improve migraine as well as tension-type headache. Antiepileptic drugs give one more good opportunity. The recognition of a psychiatric comorbidity is mandatory for an accurate management of the patient because prevents the clinicians from using any drug that might be dangerous for a mysdiagnosed psychiatric disturbance and often permits to administer medications that can efficaciously control both headache and psychiatric disorders. PMID- 16362673 TI - Genetics of cluster headache: an update. AB - Up to a few years ago, cluster headache (CH) was not thought to be an inherited disorder. However, several recent studies have suggested that genetic factors play a role in the disease. Genetic epidemiological surveys have shown that first degree relatives of CH patients are more likely to have CH than the general population. CH has been reported in some concordant monozygotic twin pairs. At present, the type and the number of genes involved in the disease are still unclear. No mutation in the CACNA1A and NOS genes was found in CH patients. Recently, we have reported a significant association between the HCRTR2 gene and the disease. The purpose of this review is to describe recent advances in the genetics of CH. PMID- 16362674 TI - New insight into trigeminal neuralgia. AB - Trigeminal neuralgia is universally considered the neuropathic facial pain most and best known in medical practice. We propose a short review on trigeminal neuralgia reporting its main clinical aspects, unsolved problems and highlighting differential diagnosis between classical and symptomatic trigeminal neuralgia. PMID- 16362675 TI - Systemic autonomic involvement in episodic cluster headache: a comparison between active and remission periods. AB - In this study we compared systemic autonomic involvement in episodic cluster headache during active and silent periods. Seventeen patients were studied with sympathetic skin response, lying to standing test, Valsalva manoeuvre, deep breathing test and orthostatic hypotension evaluation. Each of them underwent these tests during active and silent periods. Values were then compared with normal controls. Our data show a parasympathetic, but not sympathetic, involvement. Moreover, this impairment seems to be "chronic", as it persists beyond the active period. PMID- 16362676 TI - Prevalence of right-to-left shunt in patients with cluster headache. AB - Recent investigations documented that the prevalence of right-to-left shunt (RLS) in patients with migraine with aura (MA) is significantly higher than in healthy controls and similar to prevalence of RLS in young patients with cryptogenic stroke (CS). Nevertheless, little data are available in the literature about RLS prevalence in the other forms of primary headache. The aim of this study was to investigate the occurrence of RLS in patients with cluster headache (CH). We enrolled 30 consecutive patients with CH diagnosis according to the IHS criteria and 40 controls. RLS was assessed with bilateral transcranial Doppler contrast (TCDc) monitoring of middle cerebral arteries. Eleven patients (37%) resulted positive to TCDc monitoring for evaluation of RLS. These data show that the presence of RLS in this group is more prevalent than in the general population and similar to that found in MA and in CS. PMID- 16362677 TI - The temperament and character inventory in women with migraine. AB - The purpose of this work was to investigate: (1) the differences in temperament and character between 49 women with migraine and 49 controls using the Temperament and Character Inventory (TCI), and (2) the extent to which these differences were related to migraine or to the presence of comorbid depression. The migraine patients scored significantly higher than the controls in two temperament dimensions-Harm Avoidance (HA) and Persistence (P)-and significantly lower in one character dimension-Self-Directedness (SDir) (Student's t). After multiple logistic regression, the TCI P and HA dimensions were significantly associated with the presence of migraine. The HA dimension was also related to the presence of depression. Our results show that in migraine the higher HA score could be partly associated to comorbid depression while the high P dimension seems to be solely related to the presence of migraine. PMID- 16362678 TI - Headache and mood disorders. AB - The aim of the study was to estimate the occurrence of mood, anxiety and disability disorders in 300 patients affected by chronic daily headache and MOH, who were observed for a 16-month period in our centre. We monitored the patients on an interview basis, concerning the anamnestic data collection related to the pre-morbid period, information given by relatives regarding the patient's behaviour during the day, attitudes towards others, maintenance of previous interests and enjoyments, and modifications of the biological rhythm. Several tests were conducted, underlining a significant correlation between headache and mood disorders, impairment of working activity, social and family life. The study shows that patients affected by chronic daily headache and MOH present high levels of anxiety, a depressive symptomatology associated with alexithymia. Moreover, it has been discovered that anxiety and depression facilitate the onset of headache, while patients suffering from pain persistence were more vulnerable to psychiatric problems. In consideration of these results, more exhaustive evaluations relating to the psychopathological aspects in patients affected by headache are necessary. PMID- 16362679 TI - Attachment styles and headache. AB - The internal working model on attachment dimensions changes with significant emotional experiences. The purpose of this study was to evaluate if and how the internal working models correlate with primary headaches. Attachment dimensions of subjects suffering from primary headaches were studied. One hundred and fourteen subjects [68 with migraine, 23 with tension-type headache (according to ICHD-I criteria), 23 with chronic daily headache (according to Silberstein's criteria)], were studied and compared with a control group of 57 subjects (matched in sex, age and social level) not suffering from any primary headache. Attachment dimensions were investigated using the Adult Attachment Questionnaire (AAQ) and the Attachment Style Questionnaire (ASQ). Headache sufferers seem to be characterised by attachment styles of the "insecure" type. In particular they seem to feel extremely ill at ease if there is an expectation of reduction of interpersonal distance. PMID- 16362680 TI - Sleep quality, chronotypes and preferential timing of attacks in migraine without aura. AB - Clinical observations show that migraine attacks have a seasonal, menstrual and circadian timing, suggesting a role of chronobiological mechanisms and their alterations in the disease, but little experimental data exists about this issue. The aim of this study was to estimate sleep quality chronotypes and the possible circadian timing of attacks in migraneurs. One hundred patients suffering from migraine without aura according to the IHS criteria (2004), and 30 controls were enrolled. Morning and evening type subjects were more represented in migraine patients than in controls and showed a tendency towards worse sleep quality and higher disability. Forty-two percent of migraineurs presented more than 75% of their attacks at night. Morning and evening types rather than intermediate and differences between real and preferred times may represent stressors that can worsen the disease. A preferential timing for occurrence of migraine attacks during the night and early morning hours was documented. PMID- 16362681 TI - The Global Campaign (GC) to Reduce the Burden of Headache Worldwide. The International Team for Specialist Education (ITSE). AB - The social perception of headache, everywhere at low levels in industrialised countries, becomes totally absent in developing ones. Headache disorders came into the World Health Organization's strategic priorities after publication of the 2001 World Health Report. Among the leading causes of disability, migraine was ranked 19th for adults of both sexes together and 12th for females. The Global Campaign (GC) to Reduce the Burden of Headache Worldwide was planned by the major international headache organizations together with WHO in order to identify and remove those cultural, social and educational barriers recognised as responsible factors for the inadequate treatment of headache disorders worldwide. Within the GC activities, the education of the medical body will represents a central pillar. An International Team for Specialist Education (ITSE) has been created to train physicians from all over the world through the acquisition of a university level Master Degree in Headache Medicine. Once trained as headache specialists, physicians will become trainers, offering education in this field to other health care providers in their own countries. In this way they will give life to a cultural chain raising awareness locally of headache, its burden and its medical control. PMID- 16362682 TI - www.cefalea.it: the first five years. AB - The Italian headache disorders website (www.cefalea.it) was launched in 1999 by the CIRNA foundation in partnership with Al.Ce, which is a lay association and member of the World Headache Alliance. In 2004, the website registered almost 130 000 hits (+200% on the 1999 figure). The most visited parts were the sections devoted to topical issues relating to headache, the list of headache specialists, the headache glossary and the support group. This article summarises the website's first five years of activity and highlights the Internet's potential to improve headache-related decision-making, behaviour and outcomes. PMID- 16362683 TI - Headache disorders as risk factors for sleep disturbances in school aged children. AB - Several epidemiological studies have shown the presence of comorbidity between various types of sleep disorders and different headache subtypes. Migraine without aura is a sensitive risk factor for disorders of initiating and maintaining sleep (odds ratio (OR) 8.2500), and chronic tension-type headache for sleep breathing disorders (OR 15.231), but headache disorder is a cumulative risk factor for disorders of excessive somnolence (OR 15.061). This result has not been reported in the clinical literature. PMID- 16362684 TI - The prevalence of allodynia, osmophobia and red ear syndrome in the juvenile headache: preliminary data. AB - The aim of the study was to determine the frequency of clinical allodynia, osmophobia and red ear syndrome in a young population. Medical records of the children admitted for headache between 1 December 2004 and 31 March 2005 were consecutively studied. A questionnaire was used to find the prevalence of allodynia, osmophobia and red ear syndrome. We visited 96 children with headache. The range of age was 6-18 years. We classified migraine in 57%, other primary headaches in 25% and secondary headaches in about 18%. The presence of ipsilateral clinical allodynia was 14.5% in migraine, osmophobia in 20% of migraine and red ear syndrome in about 24% of migraine cases and they were absent in the other two headache groups. Our study shows that features like osmophobia, allodynia and red ear syndrome are not uncommon in migraine while they are absent in other types of headache. PMID- 16362685 TI - Chronic daily headache in a paediatric headache centre. AB - The prevalence and the clinical features of chronic daily headache (CDH) were studied in 968 children and adolescents observed during a period of one year in the Headache Centre of the Anna Meyer Paediatric Hospital of Florence. Nine hundred and forty-four patients (97.52%) had primary headache according to ICHD II, 24 subjects had secondary headache and 56 patients had CDH (5.93% of primary headaches). The mean age of subjects with CDH was higher than general (13.5 vs. 11.5 years), with a female preponderance (69.6% vs. 30.4%). According to the ICHD II, headaches were classified as chronic migraine in 10 patients (1.5.2 ICHD-II), chronic tension-type headache in 36 (2.3 ICHD-II), new daily persistent headache in 8 (4.8 ICHD-II) and 2 patients reported mixed pattern (chronic migraine+chronic tension type headache). Medication overuse was not implicated in our patients. PMID- 16362686 TI - A questionnaire-based study on prevalence and treatment of headache in young children. AB - Headache occurs in a large proportion of young students. In this study we evaluated the incidence of headache in young people, examined associated factors and described the drug therapy used. A structured questionnaire was directly administered to 2700 students of secondary schools of Catanzaro. We recorded that headache symptoms started between 9 and 12 years of age. The incidence density was higher in people living in the city (84%) and in tobacco and alcohol users than non-users (p < 0.01). The most common drugs used were non-steroidal anti inflammatory drugs. In 20% of people, this treatment induced the development of chronic headache. These data showed that morbidity from headache is often unrecognised and undertreated. Better management of headache symptoms and therapy could reduce the frequency of headache episodes, with a reduction of adverse drug reactions. PMID- 16362687 TI - Cost assessment of headache in childhood and adolescence: preliminary data. AB - Headache, and in particular migraine, is a common disturbance in childhood and adolescence. The disabling nature of headache, evident in the adult, together with its effects on family life and reduction in performance of scholastic activity, make it a disease with an elevated social economic impact. We present preliminary results of a prospective study conducted over 6 months on a population of headache sufferers in childhood and adolescence who referred to our Juvenile Neuropsychiatry Centre of the Hospital of Perugia. Our objective was to quantify the direct and indirect costs associated with juvenile headache. PMID- 16362688 TI - Post-traumatic headache from moderate head injury. AB - The onset of post-traumatic headache occurs frequently in children, where it is often caused by severe head injuries, therefore, it is part of a post-traumatic syndrome, rather than of an independent headache and in this case no cause is clearly evident. The problem, conversely, arises in post-traumatic headache after a light trauma, since it is difficult to establish the cause-effect link. We have studied PTH incidence for one year in the patients of the emergency ward of the Saint Charles of Nancy Hospital, compared to the activity of 4 Italian headache centres. At the Saint Charles of Nancy Hospital of 98 patients with PTH after a moderate head trauma, 18 had acute and 26 chronic PTH, the majority ceased after six months. In the Italian headache centres 1,656 patients were examined, of these 3.2% suffered from PTH: 25 acute, 29 chronic. These data confirm the poor evidence of PTH after a light trauma and lead to doubt of the existence of this nosological entity. PMID- 16362689 TI - Primary headache in Emergency Department: prevalence, clinical features and therapeutical approach. AB - Headache is one of the most common reported complaints in the general adult population and it accounts for between 1% and 3% of admissions to an Emergency Department (ED). The overwhelming majority of patients who present to an ED with acute primary headache (PH) have migraine and very few of them receive a specific diagnosis and then an appropriate treatment. This is due, in part, to a low likelihood of emergency physicians diagnosing the type of PH, in turn due to lack of knowledge of the IHS criteria, and also the clinical condition of the patients (pain, border type of headache, etc.) In agreement with the literature, another interesting aspect of data emerging from our experience is that few of the ED PH patients are referred to headache clinics for diagnosis and treatment, especially if they present with high levels of disability. This attitude promotes the high cost phenomenon of repeater patients that have already been admitted to the ED for the same reason in the past. This is statistically important because it involves about 10% of the population with PH. PMID- 16362690 TI - Indications for the diagnosis and treatment of acute headaches correlated with neurological pathologies. AB - The majority of patients consulting a doctor for headache are generally suffering from a primary headache. Nevertheless, the possibility of it being symptomatic, and sometimes life threatening, must always be in the forefront of the doctor's mind. Medical history and clinical examination first, and instrumental examinations afterwards, are the keys to a correct diagnosis, and the first steps for correct treatment. PMID- 16362691 TI - Management of cluster headache in the Emergency Department. AB - Although cluster headache (CH) is considered one of the most distinctive and painful primary headache disorders in clinical practice because of the brevity of each attack, its management is not always ergonomic or possible in the Emergency Department. In case of a previously competent diagnosis, the Emergency Department's team should send the patient to a headache centre where specialists in the management of CH can handle the pathology in the best way. In our headache centre we treat patients with CH attacks with a hyperbaric chamber, confirming the effectiveness of hyperbaric oxygen in CH patients. PMID- 16362692 TI - Nontraumatic headache in the Emergency Department: a survey in the province of Trieste. AB - The objective was to study the demographics, diagnostic procedures and therapies employed in order to provide guidelines to Emergency Department (ED) physicians. A six-month retrospective analysis of the records of all patients presenting with nontraumatic headache (NTH) to the EDs of the Province of Trieste was performed. Of 38,238 patients screened, 300 (0.8%) presented with NTH and 49.7% were referred to specialists. Patients were classified as having secondary headache (41.3%), primary headache (24.3%) and headache with no obvious source (NOS) (34.4%). One hundred and seventy patients were treated with mono- or polytherapy. Of 50 patients with migraine, 36 were treated with NSAIDs and 4 with triptans. 68.4% of patients were referred to a general practitioner and 31.6% were admitted. The frequency of NTH was lower than in other studies. NOS headache was frequent. Only 10% of migraineurs received triptans. Diagnostic and therapeutic guidelines for ED physicians are needed. PMID- 16362693 TI - Migraine in the Emergency Department: a psychometric study of a migraine "repeaters" sample. AB - To evaluate the influence of psychometric variables on the "repeater" phenomenon in an emergency department, 15 "repeaters" and 27 outpatient migraineurs were recruited. All patients were submitted to the Beck Depression Inventory (BDI), State and Trait Anxiety Inventory, Toronto Alexithymia Scale-20 (TAS-20), Tridimensional Personality Questionnaire and Migraine DIsability Assessment Scale (MIDAS). The "repeater" group showed higher MIDAS total scores (p=0.02) and higher scores in TAS-20 (p=0.02) than the outpatients. A higher frequency of alexithymic trait (p=0.02) and higher BDI scores (p=0.07) have also been observed in the "repeater" group than the outpatients. Alexithymia and depressive mood associated with high disability may be a specific psychosocial pattern of "repeater" migraineurs. The psychometric evaluation of this population may be important to explain the "repeaters" phenomenon. PMID- 16362694 TI - Migraine disease: evolution and progression. AB - Migraine is a complex pathology and it should be regarded as a disease evolving during the lifetime along with other comorbid conditions. Migraine susceptibility may be unmasked by exogenous substances and the occurrence of migraine attacks may change following drugs given for therapeutical purposes. The evolution of migraine should be followed up because childhood migrainous manifestations may vary over the years and an earlier diagnosis may not apply later on. PMID- 16362695 TI - A CARE: pathway in medication-overuse headache: the experience of the Headache Centre in Pavia. AB - Medication-overuse headache (MOH) is one of the headache forms that most frequently prompts patients to consult a specialist headache centre. The prevalence of this form in the general population is approximately 1-2%. Around 40% of patients seen at headache centres present with a chronic form of headache and 80% of this chronic headache patients make excessive use of symptomatic drugs. MOH shows a clinical improvement, accompanied by a reduction in the consumption of analgesic drugs, if patients are submitted to detoxification therapy. But detoxification is only the first stage in a long and complex course of care and global approach demands adequate follow-up visit to prevent early relapses. At the Headache Centre of the C. Mondino Institute of Neurology in Pavia, a course of care (CARE: ) has been developed for the complete management of patients with MOH both during Hospitalization and during the subsequent follow up period. CARE: IS designed to trace the clinical, psychopathological and pharmacological profile of MOH in the short-, medium- and long-term; to look for factors possibility predictive of relapse; to assess the direct costs linked to overuse-headache in the year leading up to and following detoxification; and to evaluate disability, in terms of working days lost, before and after detoxification. PMID- 16362696 TI - Continuity of healthcare for headache patients: a problem of communication between headache specialists and general practitioners. AB - The continuous care of headache patients, from headache centres to general practice, is a managerial problem that is still unsolved in Italy. In fact, if on the one hand patients do not usually go to headache centres because of poor information, on the other hand, if they do, they do not find their general practitioner (GP) sufficiently prepared to continue the management. In Sicily we have formed a dense network of headache centres that we will try to link on the Internet to deal with the problem of poor patients information and poor specialist consultation. We also have faced the problem of the continuous care, trying to overcome "the difficulties of communication between specialists, GPs and patients" and "the difficulties of GPs in diagnostic work", by simple instruments like the Italian version of ID-Migraine, a simple three-item questionnaire. PMID- 16362697 TI - Study on management of headache by general practitioners in South Italy. AB - The impact of migraine headaches is one of the major public health problems in several industrialised countries, with many patients reporting frequent and significant disability. Previous studies have assessed general practitioners' (GPs) perceptions towards evidence-based medicine (EBM) and its influence on health care decisions. Of 500 questionnaires distributed, responses were received from 455 for a response rate of 91%. Respondents' awareness of technical terms used in EBM indicated that only 27.2% of GPs agreed that clinical trials are needed to evaluate the efficacy of treatments and this awareness was higher in those who learned about migraine from scientific journals or continuing education courses and who attended courses on epidemiology or EBM. Training and continuing educational programmes on EBM and guidelines in terms of treatments of headache for GPs are strongly needed. PMID- 16362698 TI - Risk-benefit and cost-benefit ratio in headache treatment. AB - The cost-benefit and the risk-benefit ratios are two of the most relevant items in ongoing health organisation procedures. The choice of a new or an old therapeutic treatment depends on a number of factors and the evaluation of the cost, in terms of economics, but also in terms of quality of life and type of facilities necessary for one treatment are crucial criteria. Therefore, we have to consider in evaluating treatment strategies not only the activity of a drug in reaching the main end-points, (i.e., pain free or headache relief) but also the safety and perception of safety by patients, and the cost effectiveness, including indirect costs compared with personal and social benefits. Because it is reasonable that a subgroup of migraine patients may have a clinically progressive disorder, studies should be necessary to assess strategies for migraine treatments. PMID- 16362699 TI - Pharmacokinetics of sumatriptan in non-respondent and in adverse drug reaction reporting migraine patients. AB - Sumatriptan is a selective agonist of 5HT1 (1B/1D) receptors, which has proved to be effective and safe for the acute treatment of migraine attacks. Nevertheless, its use by migraine sufferers is still limited and some patients consider adverse reactions related to sumatriptan, especially chest symptoms, unacceptable even if not serious. Moreover, in clinical trials, almost one third and one sixth of patients, respectively, fail to experience headache relief either after oral or after subcutaneous sumatriptan administration. Our aim was to verify whether differences in sumatriptan pharmacokinetics could explain non-response and/or adverse drug reactions. Sumatriptan levels were determined by HPLC with electrochemical detection. Pharmacokinetic parameters were calculated using a computer program (PK Solutions 2.0; non compartmental Pharmacokinetics Data Analysis). After oral administration, sumatriptan is rapidly absorbed and sometimes displays multiple peaks of plasma concentration. This "multiple peaking" gives rise to considerable inter-subject variability in the time of reaching maximum plasma concentration. Pharmacokinetic parameters of sumatriptan, both after oral and subcutaneous administration, were similar in the three patient groups. Blood pressure and heart rate did not show any significant differences between groups. Pharmacokinetic parameters and bioavailability of sumatriptan did not seem to be correlated either to the lack of efficacy or the appearance of side effects. These results could depend on the limited number of patients studied. PMID- 16362700 TI - Weight variations in the prophylactic therapy of primary headaches: 6-month follow-up. AB - We conducted a study on 367 patients (86% female, 14% male; mean age 37+/-15 years) suffering from migraine with and without aura and chronic tension-type headache to evaluate the incidence of weight gain, an undesirable side effect observed during prophylactic therapy in primary headaches. Patients treated with amitriptyline (20 and 40 mg), pizotifen (1 mg), propranolol (80-160 mg), atenolol (50-100 mg), verapamil (160-240 mg), valproate (600 mg) and gabapentin (900-1200 mg) were evaluated after a period of 3 and 6 months. In particular, 89 patients were assessed (78% female, 22% male) at 6 months, of whom 10 were in treatment with amitriptyline 20 mg, 19 with amitriptyline 40 mg, 7 with pizotifen (1 mg), 13 with propranolol (80-160 mg), 4 with verapamil (160 mg), 10 with valproate (600 mg), 15 with atenolol (50 mg) and 11 with gabapentin (900-1200 mg). The control group consisted of 97 patients with migraine (79% female, 21% male; mean age 35+/-16 years) without indication for prophylactic therapy. Weight variations >or=1 kg were considered. After 6 months of therapy, the percentage of patients with weight gain was 86% with pizotifen (6/7; mean weight increase 4.4+/-2.5 kg), 60% with amitriptyline 20 mg (6/10; 3.1+/-1.6), 47% with amitriptyline 40 mg (9/19; 5.4+/-2.7), 25% with valproate 600 mg (2/8, 3.0+/-2.8 kg), 25% with verapamil (1/4, 2.5 kg), 20% with atenolol (3/15, 1.7+/-0.6 kg), 9% with gabapentin (1/11, 1.5 kg) and 8% with propranolol (1/13; 6 kg). We conclude that propranolol, gabapentin, atenolol, verapamil and valproate affect body weight in a modest percentage of patients at 6 months. A greater mean weight gain at 6 months was found in patients treated with pizotifen, amitriptyline, and, in one patient out of 13, with propranolol. PMID- 16362701 TI - Headache and botulinum toxin. AB - The authors discuss clinical and international experience about botulinum toxins (BTX types A and B) in headache treatment. Data from literature suggest good results for the treatment of tension-type headache, migraine and chronic tension type headache. In the present paper mechanisms of action and injection sites will also be discussed. PMID- 16362702 TI - Prevalence of patent foramen ovale in a large series of patients with migraine with aura, migraine without aura and cluster headache, and relationship with clinical phenotype. AB - A relationship between migraine and patent foramen ovale (PFO) has been observed in relatively small series of patients so far. Furthermore, the exact mechanism underlying such an association remains unknown. In the present study we determined the prevalence of PFO by contrast-enhanced transcranial Doppler (TCD) in a group of 260 patients with migraine with aura (MA+), 74 patients with migraine without aura (MA-), and 38 patients with cluster headache (CH). One hundred-sixty-one MA+subjects (61.9%), 12 MA-subjects (16.2%), and 14 CH-subjects (36.8%) were PFO-carriers. The association was independent on the frequency of migraine attacks and complexity of aura. Finally, among the 15 patients who had a history of at least one migraine attack occurring during a Valsalva maneuver only one subject turned out to be PFO-carrier. Our findings confirm previous observations of a link between MA+, CH, and PFO. They also suggest that such an association is independent on migraine clinical phenotype and is probably unrelated to the pathogenic mechanism of paradoxical embolism. PMID- 16362703 TI - Preventing chronicity of migraine. AB - Chronic migraine and transformed migraine are conditions with a progression from episodic to chronic headache, a disabling stage. During attack, cutaneous allodynia frequently occurs: it reflects sensitisation of the central neurons of the trigeminovascular system. Early triptan therapy (prior to the development of central sensitisation) may protect from the chronicisation of migraine. In addition, early recognition of non-headache changes in neurologic function between episodes of headache offers a sensitive indicator of headache transformation. Attack frequency is the stronger predictor for migraine progression: prophylactic agents could be administered to patients with a high number of attacks. Medication overuse is the most important iatrogenic risk factor for the acceleration of disease and it must be prevented; other important risk factors are female sex, obesity and stressful life events. PMID- 16362704 TI - Preliminary results of a withdrawal and detoxification therapeutic regimen in patients with probable chronic migraine and probable medication overuse headache. AB - Chronic migraine (CM) is an invalidating condition affecting a significant population of headache sufferers, frequently associated with medication overuse headache (MOH). Controlled trials and guidelines for the treatment of MOH are currently not available. We studied the efficacy of a therapeutic regimen for the withdrawal of the overused drug and detoxification in a sample of patients suffering from probable CM and probable MOH during admission in eight hospitals of Piemonte-Liguria-Valle d'Aosta. Fifty patients, 42 females (84%) and 8 males (16%), mean age at observation 50.66+/-13.08 years, affected by probable CM and daily medication overuse following IHS diagnostic criteria were treated as inpatients or in a day hospital. Headache index (HI) and daily drug intake (DDI) were used for evaluating the severity of headache and medication overuse. The patients were treated by abrupt discontinuation of the overused drug and by a therapeutic protocol including i.v. hydration, dexamethasone, metoclopramide and benzodiazepines for 7-10 days. Prophylactic medication was started immediately after admission. Analgesics or triptans were used under medical control only in cases of severe rebound headache. Diagnostic protocol included routine blood tests (at admission and at discharge), dosage of B12 and folic acid. Patients underwent follow-up controls one, three and six months after discharge. The initial diagnosis was probable CM in almost all patients included in the study (41 patients); in nine patients the diagnosis was not specified (coded only as CDH). The overused medications were simple analgesics in 17 cases (34%), combination analgesics in 19 cases (38%), triptans alone or with analgesics in 13 cases (26%) and ergotamine in 2 cases (4%). We collected data from 39 patients at first follow-up (1 month), 32 after 3 months and 14 after 6 months. Mean HI was 0.91 at admission, 0.22 at discharge, 0.38 after 30 days, 0.46 after 3 months and 0.48 after 6 months. Mean DDI was 2.80 at admission, 0.39 at discharge, 0.41 after 1 month, 0.52 after 3 months and 0.59 after 6 months. These results are on average positive and tend to remain stable with time. Although preliminary and obtained on a limited number of patients at 6-month follow-up, our results seem to be encouraging about the use of the proposed therapeutic protocol. PMID- 16362705 TI - Psychiatric comorbidity and chronicisation in primary headache. AB - The frequent association between primary headaches and psychiatric disorders is consistently reported in the literature. There is increasing evidence that a bi directional relationship links these somatic conditions to psychopathological events. Prospective studies show that several psychiatric disorders are severe risk factors for both the onset and chronicisation of primary headache, and for a long time it has been suspected that headache triggers psychiatric disorders, mostly of affective nature, and affects both their course and outcome. Researchers are actively involved in investigating the biological basis of such a relationship while clinicians still need to strengthen their interest in psychiatric comorbidity of their primary headache patients to improve clinical outcome and to prevent chronic evolutions. PMID- 16362706 TI - Preventive non-pharmacological treatment and nitric oxide in chronic migraine. AB - In chronic migraine the central sensitisation and the changes of regional cerebral blood flow are mediated by nitric oxide (NO) and oxygen free radicals. Biofeedback is considered a preventive non-pharmacological treatment decreasing migraine attacks. We investigated whether biofeedback effectiveness is related to relaxation processes and its influence on oxidative stress. The Migraine Disability Assessment Score (MIDAS) and serum NO stable metabolites (NO(x)) were evaluated in 20 patients with chronic migraine before and at the end of biofeedback sessions. MIDAS score was lower after biofeedback than that reported before treatment. NO(x) serum levels were higher after biofeedback than those measured before starting treatment. Thus, the effectiveness of biofeedback is related to a muscular relaxation and to its influence on NO bioavailability in patients with chronic migraine. PMID- 16362709 TI - Lifting The Burden: the global campaign to reduce the burden of headache worldwide. AB - Lifting The Burden envisions a future world in which headache disorders are recognized everywhere as real, disabling and deserving of medical care to which all who need it have access without artificial barriers. PMID- 16362710 TI - Migraine aura without headache compared to migraine with aura in patients with affective disorders. AB - The characteristics of psychiatric comorbidity in migraine have been studied in migraine with aura (MA) and migraine without aura (MO). Little information is available concerning patients with migraine aura without headache. In a study of 201 patients with major affective disorders (DSM-IV) we have described the clinical characteristics of patients with these three sub-types of migraine (IHS criteria) and compared the MA and migraine aura without headache groups. Compared to patients having MA (n=57), the group with migraine aura without headache (n=18) had a higher age of onset of migraine (28.5 vs. 19.2, p=0.001), significantly lower prevalences of affective temperaments (28% vs. 56%, p=0.036), suicide attempts (17% vs. 53%, p=0.013) and Raynaud's syndrome (0% vs. 25%, p=0.017). These results indicate that there seem to be differences in the clinical characteristics found in patients with migraine with aura when compared to those having the migraine subtype without a headache phase. This may convey new information concerning the comorbid expression of migraine and affective disorders or concerning the processes that differentiates the migraine types with and without a subsequent pain attack. PMID- 16362711 TI - Effect of preventive treatment on health-related quality of life in episodic migraine. AB - The purpose of our study was to prospectively evaluate the impact of preventive drug treatment on the quality of life of patients with episodic migraine (EM). Quality of life evaluations can enhance traditional measures of therapeutic efficacy. Thirty-five consecutive EM without aura patients attending a tertiary care unit (Batatais Headache Clinic) entered the study. They were given a prescription for preventive medication and completed the validated Portuguese version of the generic instrument Short Form-36 (SF-36) questionnaire. Six months later, patients were given another SF-36. We compared the pre-treatment and post treatment scores for the SF-36 and analysed the headache diaries. The mean age of the 35 patients (32 women and 3 men) was 40.2 years (range 18-60 years). All 35 patients completed the pretreatment and follow-up SF-36. The pre-treatment and post-treatment mean frequency of migraine attacks was 9.16 and 2.4 crises per month, respectively (p<0.05). After the six months of preventive treatment, six of the eight domains evaluated by the SF-36 (role physical, bodily pain, general health, vitality, social function and mental health) showed statistically significant (p<0.05) improvement. The preventive treatment of migraine leads to a significant improvement in the quality of life of patients with EM. This improvement was measurable by the general quality of life instrument SF-36. PMID- 16362712 TI - Anger and emotional distress in patients with migraine and tension-type headache. AB - The objective was to evaluate the prevalence and the characteristics of anger and emotional distress in migraine and tension- type headache patients. Two hundred and one headache patients attending the Headache Center of the University of Turin were selected for the study and divided into 5 groups: (1) migraine, (2) episodic tension-type headache, (3) chronic tension-type headache, (4) migraine associated with episodic tension-type headache and (5) migraine associated with chronic tension-type headache. A group of 45 healthy subjects served as controls. All the subjects completed the State-Trait Anger Expression Inventory, the Beck's Depression Inventory and the Cognitive Behavioral Assessment. Anger control was significantly lower in all headache patients (p<0.05) except in migraineurs. Patients with migraine and tension-type headache showed a significantly higher level of angry temperament and angry reaction (p<0.05). In addition, chronic tension-type headache and migraine associated with tension-type headache patients reported a higher level of anxiety (p<0.05), depression (p<0.001), phobias (p<0.001) and obsessive-compulsive symptoms (p<0.01), emotional liability (p<0.001) and psychophysiological disorders (p<0.001). Our study shows that chronic tension-type headache and migraine associated with tension-type headache patients present a significant impairment of anger control and suggests a connection between anger and the duration of headache experience. PMID- 16362713 TI - Diagnostic lumbar puncture. Comparative study between 22-gauge pencil point and sharp bevel needle. AB - Post-lumbar puncture headache is a frequent clinical problem. Needle design is expected to reduce post-puncture headache. In this study, we compared two different lumbar puncture needle designs in diagnostic lumbar puncture and analysed post-dural puncture headache (PDPH) and social and economical harm associated with the diagnostic lumbar puncture procedure. This prospective, controlled study consisted of 80 consecutive adult patients requiring elective diagnostic lumbar puncture due to various neurological symptoms. Lumbar puncture was completed either with Spinocan 22 G sharp bevel needle or Whitacre 22G pencil point needle. Patients were asked about previous headache symptoms and pain provoked by puncture. One week after the lumbar puncture all patients were interviewed by telephone and occurrence and type of headache, headache intensity, medication and frequency of impairment in activities of daily living were asked. Need for epidural blood patch was also recorded. Thirty-three of 78 (42%) patients experienced headache after diagnostic lumbar puncture and in 26 (33%) the headache could be classified as PDPH. There were no statistically significant differences between needle types in the frequency of common headache, PDPH, puncture pain intensity, need for epidural blood patch or sick leave. Also, there were no other complications except local back pain or headache. In this study, the needle design did not affect the frequency of PDPH. Also, PDPH was common, occurring in 33% cases and caused a considerable amount of disturbance in daily activities. Seeking help for this condition was insufficient and only part of these PDPH patients were treated with epidural blood patch. PMID- 16362714 TI - Treatment satisfaction with zolmitriptan nasal spray for migraine in a real life setting: results from phase two of the REALIZE study. AB - In phase one of the REALIZE study, zolmitriptan nasal spray demonstrated a significant headache response from 10 min post-dose and total symptom relief from 30 min post-dose. The objective of phase two was to investigate patients' dosing patterns, satisfaction and preference following open-label treatment with the nasal spray. Up to 3 attacks were treated. The ITT population consisted of 851 patients. The median time from onset of symptoms to treatment was 1 h 15 min (primary endpoint). Most patients reported being satisfied or very satisfied with zolmitriptan nasal spray (75.7%). Furthermore, the majority of patients would be willing to use zolmitriptan nasal spray in the future (59.8%) and preferred zolmitriptan nasal spray over previous therapies (57.8%). Zolmitriptan nasal spray was well tolerated. Most patients were satisfied with zolmitriptan nasal spray, were willing to continue using it and preferred it to previous therapies. PMID- 16362715 TI - Clinic-based study of family history of vascular risk factors and migraine. AB - The objective was to evaluate the presence of a positive family history (FH) of vascular risk factors between patients with migraine with aura (MA) and migraine without aura (MO), and in chronic migraine (CM) compared to other headache types. As migraine patients are typically too young to have developed vascular events, studying older relatives of migraine patients may be a practical means of evaluating associations between vascular risk factors and migraine. A cross sectional study of a clinic-based sample of adults with migraine headache was carried out at the University of Utah. Predictor variables comprised first or second degree relatives with stroke, hypertension, diabetes or hypercholesterolaemia. Outcome measures comprised diagnosis of MA, MO or CM according to the revised International Headache Society criteria. There was no significant difference in FH of vascular risk factors in MA compared to MO (adjusted OR 1.04, 95% CI 0.61-1.78). CM was associated with a decreased risk of FH of stroke (OR=0.11, 95% CI 0.02-0.87, p=0.036). There was no significant difference in FH of vascular risk factors in MA patients compared to MO. CM patients were more likely to have a negative FH of stroke compared to other headache types, suggesting that CM is likely a neuronal disease rather than a vascular one. PMID- 16362716 TI - Warfarin as a therapeutic option in the control of chronic cluster headache: a report of three cases. AB - Chronic cluster headache remains refractory to medical therapy in at least 30% of those who suffer from this condition. The lack of alternative medical therapies that are as effective as, or more effective than, lithium carbonate makes new therapies necessary for this highly disabling condition. Based on a previous report, we gave oral anticoagulants to three patients with chronic cluster headache. Two of them remained cluster headache-free while taking warfarin. In the third patient, the use of warfarin for three weeks initially increased the frequency and intensity of cluster headache attacks but subsequently induced a prolonged remission. In spite of the paucity of data available, oral anticoagulation appears to be a promising therapy for chronic cluster headache. PMID- 16362717 TI - Guidelines for the management of headache in primary care: are they being used? AB - The uptake of guidelines for the management of headache in primary care was assessed using surrogate measures in patients with headache referred to secondary care (number of prior general practitioner attendances and medications prescribed). The results suggest that headache guidelines are seldom, if ever, applied in this geographical location. PMID- 16362718 TI - "The Headache Week": a useful tool to highlight "invisible" migraineurs. PMID- 16362719 TI - Muscular dystonia and athetosis in six patients with congenital nephrotic syndrome of the Finnish type (NPHS1). AB - Congenital nephrotic syndrome of the Finnish type (NPHS1, CNF) is an autosomal recessively inherited disease occurring due to mutations in the nephrin gene (NPHS1). Two main Finnish mutations exist: Fin-major and minor, which both cause a lack of nephrin and absence of the slit diaphragm between the podocytes. This leads to severe proteinuria, nephrotic syndrome and infections, and without dialysis or renal transplantation, death in infancy. Between 1984 and 2003, six (8.6%) of the 70 NPHS1 patients diagnosed at our institution had, in addition to their renal disease, similar neurological symptoms. All six showed a severe dyskinetic cerebral palsy-like syndrome with dystonic features, athetosis and a hearing defect. The neurological symptoms became apparent during their 1st year of life and were diagnosed before 11 months of age. MRI showed increased signal intensity in T2-weighted images in the globus pallidus area. No mitochondrial gene mutations explaining the neurological symptoms were found, nor did external neurological complications explain them when compared with 29 NPHS1 control patients. Four children died at an early age: two during dialysis and two shortly after renal transplantation. Two are still alive with a functioning graft. Both have severe motor defects, but are mentally active and social. PMID- 16362720 TI - A case of atypical hemolytic uremic syndrome with a transient decrease in complement factor H. AB - We report a case of sporadic atypical hemolytic uremic syndrome (HUS) with a transient decrease in complement factor H. Referred for hemolysis and azotemia without diarrhea prodrome, this 31-month-old boy showed a decreased complement 3 (C3) and complement factor H (FH) level. However, the factor H gene (HF1) mutation was missing. After the hemolysis was controlled with plasma infusion, the C3 and FH levels recovered. The patient's renal function fully recovered and remained normal, and there was no recurrence of the HUS. PMID- 16362721 TI - Outcome of isolated antenatal hydronephrosis: a systematic review and meta analysis. AB - Idiopathic antenatal hydronephrosis (IAHN), defined as antenatal hydronephrosis not associated with other morphologic renal tract abnormalities, is the most common abnormality detected by antenatal ultrasound. We performed a systematic review and meta-analysis to determine the outcome of IAHN. We screened three databases and bibliographies to identify English-language original peer-reviewed papers that reported serial postnatal renal ultrasonography in children with IAHN. Patients who stabilized and/or improved were extracted and pooled according to the individual grading systems used by each study. A systematic analysis of data extracted from 25 articles revealed overall resolution of pelviectasis in milder cases of IAHN (Society of Fetal Urology [SFU] grade 1-2; anterior posterior pelvic diameter [APPD]<12 mm). In contrast, IAHN of higher severity (grades 3-4; APPD>12 mm) resolved with a lower frequency. Meta-analysis of data extracted from seven papers showed stabilization of pelviectasis in 98% of patients with grades 1-2 (95% confidence interval [CI] 0.93-1.0; p =0.0008) and in 51% of patients with grades 3-4 (95% CI 0.34-0.68; p <0.00001). Grades 1-2 pelviectasis was five times more likely to stabilize than grades 3-4 pelviectasis (odds ratio [OR] 4.69; 95% CI 1.73-12.76; p =0.002). We conclude that in patients with IAHN and lesser degrees of pelvic dilatation, pelvic diameter decreases to the normal range or does not worsen with the vast majority of patients. Further studies are needed to define outcomes, particularly in more severe forms of IAHN. PMID- 16362722 TI - Intracellular cytokines in peripheral blood leucocytes in children with chronic renal failure. AB - We have previously shown that children with mild renal impairment show significant changes in leucocyte subsets and circulating cytokines, indicating that these patients show an increased inflammatory state. We hypothesised that measurement of intracellular cytokine production by lymphocytes and monocytes would more precisely define the immunological mechanism associated with the inflammatory state in children with pre-dialytic chronic renal failure. Blood was collected from children with chronic renal failure (CRF) who were not yet on dialysis and an age-matched control group. Leucocyte subsets and intracellular cytokine production were determined using flow cytometry. Children with CRF showed increased production of interleukin (IL)-12 by monocytes accompanied by decreased production of interferon (IFN)-gamma and increased production of IL-4 by T cells. There were no significant changes in the production of IL-8, IL-10, IL-6, IL-1alpha or tumour necrosis factor (TNF)-alpha by monocytes or in IL-2 or TNF-alpha production by T cells. There were no significant differences in total white cell count or lymphocyte count. There was a significant decrease in both B and NK cells. This study examines intracellular cytokine production in children with CRF in detail. It is the first to show that children with relatively mild renal failure display significant immunological changes of lymphocyte subsets and leucocyte cytokine production. These data provide a more accurate understanding of the immunological changes that may contribute to the clinical manifestations and progression of the disease. PMID- 16362723 TI - Interaction of kinesin motors, microtubules, and MAPs. AB - Kinesins are a family of microtubule-dependent motor proteins that carry cargoes such as vesicles, organelles, or protein complexes along microtubules. Here we summarize structural studies of the "conventional" motor protein kinesin-1 and its interactions with microtubules, as determined by X-ray crystallography and cryo-electron microscopy. In particular, we consider the docking between the kinesin motor domain and tubulin subunits and summarize the evidence that kinesin binds mainly to beta tubulin with the switch-2 helix close to the intradimer interface between alpha and beta tubulin. PMID- 16362724 TI - TGF-beta1 favors the development of fast type identity during soleus muscle regeneration. AB - Transforming growth factor-beta1 (TGF-beta1) is known to be expressed in the environment of developing fast muscle fibres during ontogenesis. In the present study, we have examined effects of administration of either TGF-beta1 or neutralizing TGF-beta1 antibody on the induction of fast type phenotype in regenerating skeletal muscles in rats. Expressions of fast and slow myosin heavy chain (MHC) isoforms were studied using protein electrophoresis, at 3 and 6 weeks after myotoxic treatment. Muscle contractile properties were also measured in situ. The results have shown that a single injection of TGF-beta1 into the regenerating slow soleus muscle increased the expression of fast MHC-2x/d and MHC 2a and decreases that of slow MHC-1 (P<0.05). Moreover, it reduced the degree of tetanic fusion during contraction (P<0.05). Conversely, injection of neutralizing antibody against TGF-beta1 into the regenerating fast EDL muscle increased the expression of MHC-2a and MHC-1 (P<0.05). In conclusion, when the slow muscle was regenerating in the presence of an increased level of TGF-beta1, it induced a shift to a less slow MHC phenotype and contractile characteristics. Conversely, neutralization of TGF-beta1 in the regenerating fast muscle induced a shift to a less fast MHC expression. Together these results suggest that TGF-beta1 influences some aspects of fast muscle-type patterning during skeletal muscle regeneration. PMID- 16362725 TI - Long-term anatomical and functional assessment of trans-vaginal cystocele repair using a tension-free polypropylene mesh. AB - We report a case series of 63 women with cystocele who underwent the same trans vaginal procedure between October 1999 and October 2002. The polypropylene mesh (GyneMesh, Gynecare, Ethicon, France) was placed from the retropubic space to the inferior part of the bladder in a tension-free fashion. Patients were followed up for 24 to 60 months, with a mean follow-up of 37 months. Fifty-five patients returned for follow-up (87.3%). At follow-up, 49 women were anatomically cured (89.1%), five women had stage 2 anterior vaginal wall prolapse (9.1%), and one had a recurrent stage 3 (1.8%). Functional results and sexual function were also investigated. Fifty-three women had significant improvement in their quality of life (96.4%). There were a total of three cases of local pain around a mesh shrinkage (5.5%) and five vaginal erosions of the mesh (9.1%). Four out of 24 patients had dyspareunia (16.7%). In conclusion, the vaginal repair of anterior vaginal wall prolapse reinforced with a polypropylene mesh was efficient at 2 to 5 years follow-up. However, the first generation of polypropylene mesh we used was responsible for high rates of local complications and dyspareunia. Therefore, the polypropylene mesh has to be improved (lower weight) and the technique has to be documented by a randomized controlled trial before we could recommend its use in clinical practice. PMID- 16362726 TI - Positive regulation of hepatic carnitine palmitoyl transferase 1A (CPT1A) activities by soy isoflavones and L-carnitine. AB - BACKGROUND: Genistein increases CPT1A, a rate-limiting enzyme in the beta oxidation pathway, enzyme activity by increasing CPT1A transcription in HepG2 cells and, consequently, suppresses high fat induced obesity in C57BL/6J mice. Genistein and daidzein are the most abundant isoflavones in soy. AIM OF STUDY: To investigate the effect of co-treatment of genistein and L-carnitine on CPT1A enzyme activity and to determine whether daidzein also increases CPT1A activity and to establish a cell line that can be used to screen chemicals to regulate CPT1A transcription. METHODS: The enzyme activities of CPT1A were determined after HepG2 cells were incubated with 10 microM genistein or 10 microM daidzein or 1 mM L-carnitine or in combination with 10 microM genistein and 1 mM L carnitine or in combination with 10 microM daidzein and 1 mM L-carnitine. The mRNA expression levels of CPT1A were determined by real time PCR method after HepG2 cells were incubated with 10 microM genistein or 10 microM daidzein. A suggested CPT1A promoter region was cloned from human genomic DNA and the CPT1A promoter-luciferase reporter gene construct was made, and the promoter-reporter gene construct was transfected into human hepatoma cell line Huh7. RESULTS: The enzyme activity of CPT1A was at least 2.3- fold higher in L-carnitine and genistein co-treated HepG2 cells than either single-agent treated cells. Daidzein also significantly increased the mRNA expression of CPT1A as well as the enzyme activity of CPT1A. A stable Huh7 cell line, which was selected after Huh7 cells were transfected with CPT1A promoter luciferase reporter gene construct, was characterized by confirming that luciferase activity of the cell line can be regulated by genistein and daidzein as well as clofibrate, a well-known CPT1A mRNA up-regulating drug. CONCLUSIONS: Genistein and daidzein can up-regulate CPT1A enzyme activity through up-regulation of CPT1A transcription. Co-treatment of L-carnitine and genistein additively increases CPT1A enzyme activity in HepG2 cells. A stable Huh7 cell line transfected with the CPT1A promoter luciferase reporter gene was established and characterized. PMID- 16362728 TI - [Primary electrical heart diseases]. PMID- 16362729 TI - [Molecular basis of primary electrical heart diseases]. AB - The last decade has seen rapid progress in our understanding of the molecular basis of arrhythmias, particularly concerning hereditary arrhythmia syndromes. This has led to significant improvement regarding differentiation, risk stratification and therapy in these patients and their families. However, there is mounting evidence that the knowledge obtained by studying these rare monogenic disorders will also enable us to dissect the molecular mechanisms underlying polygenetic and multi-factorial arrhythmias that are by far more common in clinical practice. The goal of this review is to give a brief overview of current knowledge on the molecular basis of primary electrical heart diseases. A focus is on the long QT syndrome. PMID- 16362730 TI - [Electrical heart diseases--therapy during childhood and adolescence]. AB - With increasing experience, radiofrequency catheter ablation of tachyarrhythmia substrates has become first choice therapy for children >4 years of age with recurrent tachyarrhythmia. In younger patients, the risks associated with the procedure (typically procedure-related AV block or possible coronary artery damage) have to be weighed against the natural history of the tachyarrhythmia substrate, and the degree of control achieved with pharmacologic agents. Ablation for postoperative arrhythmias is more complicated, and associated with lower success rates and a higher rate of recurrence (of the same or a new tachyarrhythmia) despite acute procedural success. In this setting, catheter ablation has to be considered in conjunction with further surgery or the use of a defibrillator as a backup device to prevent arrhythmia-related sudden death. Also in inherited arrhythmias as in long QT syndrome and Brugada syndrome, implantable defibrillators have to be considered as a possible therapeutic option for patients with a higher risk for sudden cardiac death, irrespective of age. PMID- 16362731 TI - [Primary electrical heart disease in adulthood--electrophysiological findings and therapy]. AB - Sudden cardiac death accounts for 100,000 victims in Germany per year. Predominantly, patients with structural heart disease such as coronary artery disease or dilated cardiomyopathy are affected. However, approximately 5-10% of sudden deaths hit patients without structural disease of the heart. The proportion of young patients (< 40 years of age) in this group is even higher (10 20%). In younger patients significantly more diseases like hypertrophic cardiomyopathy, arrhythmogenic right ventricular dysplasia and primary electrical diseases of the heart could be observed such as long QT syndrome, short QT syndrome, Brugada syndrome and catecholaminergic polymorphic ventricular tachycardia. The primary electrical diseases are different concerning their electrocardiographical pattern, clinical triggers of arrhythmias, results of invasive diagnostics and therapy. Meanwhile, molecular genetic screening can reveal specific mutations of ion channels and can identify consecutive functional defects. The significance of programmed ventricular stimulation is at present unclear concerning risk stratification in patients with Brugada syndrome and short QT syndrome and of no significance in long QT syndrome and catecholaminergic polymorphic ventricular tachycardias. The implantable cardioverter defibrillator is the therapy of choice in most symptomatic patients. With increasing knowledge as a result of sophisticated molecular genetic screening, identification of underlying ion channel defects and new details of the mechanisms of arrhythmogenesis, a potential genotype-guided therapy will gain more importance in the future. PMID- 16362732 TI - [Right ventricular tachyarrhythmias--diagnostics and therapy]. AB - Ventricular tachyarrhythmias originating from the right ventricle frequently occur in young, apparently healthy patients with rare underlying cardiac diseases. Among these are arrhythmogenic right ventricular cardiomyopathy (ARVC), idiopathic right ventricular outflow-tract tachycardia (RVOVT), and Brugada syndrome (BrS). All harbor the risk of sudden cardiac death, whereas they differ substantially as to diagnosis, therapy and prognosis. This is the reason why detailed investigations are an essential prerequisite for further efficient individualized management strategies which are mainly directed to prevent sudden cardiac death and to minimize the risk of arrhythmia recurrences in affected patients, respectively. Both antiarrhythmic drug therapy, catheter ablation, and the implantation of an automatic cardioverter defibrillator may, therefore, be a first-line therapeutic option in tailored treatment regimens. This review is a summary of the available literature on pathogenesis, diagnosis, treatment, and prognosis of such diseases associated with right ventricular tachyarrhythmias. PMID- 16362733 TI - [Atrial flutter after orthotopic heart transplantation due to recipient-to-donor transatrial conduction]. AB - A few weeks after orthotopic heart transplantation, a male adolescent developed atrial arrhythmias of the donor heart due to an atypical recipient atrial flutter with a recipient-to-donor transatrial conduction resulting in an absolute arrhythmia. Under medication with propafenone, the atrial flutter of the donor heart could be terminated with cardioversion. PMID- 16362734 TI - Transversal crista terminalis conduction suggests ineffective bidirectional isthmus block. AB - Catheter ablation of the posterior isthmus is an effective tool to cure typical atrial flutter. In some cases, however, bidirectional block cannot be obtained despite extensive RF applications. Anatomic obstacles or abnormalities are thought to be the most common reasons for failed or prolonged procedures. We present a case of recurrent typical atrial flutter that seemed to be refractory to all ablation attempts in the region of the posterior isthmus although no anatomic abnormalities could be detected. Despite extensive RF application, bidirectional conduction was unchanged. Using a novel noncontact mapping system (En-Site 3000) the existence of a fast conducting gap in the region of the inferior terminal crest was revealed. Rapid conduction over this gap to the opposite side of the isthmus led to the impression that bidirectional isthmus block was not established. As a result no further RF applications were necessary because isthmus block was complete at that time. This is the first time that transverse conduction across the terminal crest could be detected by this novel noncontact mapping system masquerading as unchanged bidirectional isthmus conduction. PMID- 16362735 TI - [Cardiac resynchronization therapy (CRT) and anodal stimulation]. AB - We describe a biventricular stimulation mode to pace both ventricles with a single electrical stimulus between the tip of the left ventricular electrode and the tip of the right ventricular electrode: one ventricle is stimulated cathodal the other anodal. PMID- 16362736 TI - [Pacemaker--EKG--Quiz]. PMID- 16362764 TI - Simo S. Oja--amino acids all along as building blocks of brain and life. PMID- 16362765 TI - Cerebrocellular swelling in the presence of uraemic guanidino compounds: ameliorative effects of taurine. AB - Cell volumes (equilibrium non-inulin spaces) have been measured in slices of rat cerebral cortex incubated in the presence of uraemic guanidino compounds. Of 5 guanidino compounds tested, all but one caused significant cell swelling. This was most pronounced for guanidinosuccinic acid (GSA, 40 micromol/l)(+22%) and guanidine hydrochloride (G, 3 micromol/l)(+13%). Swelling was reduced by taurine in a dose-dependent manner, being completely abolished at 20 mmol/l. Swelling was also abolished by the antioxidants ascorbic acid (0.4 mmol/l) and butylated hydroxytoluene (0.5 mmol/l), the free radical scavenger N-acetyl-L-cysteine (10 mmol/l) and the lipid peroxidase inhibitor desmethyl tirilazad (100 micromol/l). The remission of swelling by 20 mmol/l taurine was reduced by 50% by the taurine transport inhibitor guanidinoethylsulphonate (GES, 1 mmol/l). This figure was not significantly altered when the concentration of GES was increased to 10 mmol/l. It was also reduced by 45% by the GABAA receptor antagonist bicuculline (100 micromol/l). It was completely abolished when both GES and bicuculline were present. It is suggested that guanidino compounds result in cells undergoing oxidative-nitrosative stress, and that taurine protects against the resultant cell swelling by 2 mechanisms One (intracellular) requires taurine transport and depends on its role as an antioxidant, with lipid peroxidation being probably a significant factor. The other (extracellular) is associated with activation of GABAA receptors. PMID- 16362766 TI - Evidence for a reduction of coupling between GABAA receptor agonist and ionophore binding sites by inorganic phosphate. AB - [35S]TBPS binding to the GABAA receptor ionophore binding site is anion dependent. Using autoradiography on rat brain sections, we show that permeabilities of anions through the receptor channel correlate with their efficiencies to promote basal [35S]TBPS binding. Phosphate made an exception as it induced more binding than expected from its permeability. Well-permeable anions (chloride, nitrate, formate) allowed [35S]TBPS binding to be effectively displaced by 1 mM GABA, whereas low-permeable anions (acetate, phosphate, propionate) markedly prevented this GABA effect, especially in the thalamus, the transition from the high to the low GABA effect being between formate and acetate. In the presence of phosphate, GABA enhanced [3H]flunitrazepam binding to benzodiazepine site of recombinant alpha1beta2gamma2 receptors with the same efficacy but lower potency as compared to the presence of chloride, whereas [35S]TBPS binding was abnormally modulated by GABA. These results suggest that inorganic phosphate affects coupling between agonist and ionophore sites in GABAA receptors. PMID- 16362767 TI - Taurine and zinc modulate outgrowth from goldfish retinal explants. AB - Taurine and zinc, highly concentrated in the retina, possess similar properties in this structure, such as neuro-protection, membrane stabilization, influencing regeneration, and modulating development, maybe by acting in parallel or as interacting agents. We previously demonstrated that there are some correlations between taurine and zinc levels in hippocampus, dentate gyrus and retina of the developing rat. In the present study we evaluate the possible effects of taurine and zinc on outgrowth from goldfish retinal explants. The optic nerve was crushed 10 days before plating and culturing retinal explants in Leibovitz medium with 10% fetal calf serum and gentamicin. Neurites were measured with SigmaScanPro after 5 days in culture. Taurine (HPLC) and zinc (ICP) concentrations were determined in the retina between 1 and 180 days after crushing the optic nerve. Zinc sulfate (0.01-100 microM), N,N, N',N'-tetrakis (pyridylmethyl) ethylenediamine (TPEN, 0.1-5 nM) and diethylenetriamine penta-acetic acid (DTPA, 10-300 microM), intracellular and extracellular zinc chelators, respectively, were added to the medium. TPEN was also injected intraocular (0.1 nM). Combinations of them were added with taurine (1-16 mM). Taurine concentrations were elevated in the retina 72 h after the crush, but were normalized by 180 days, those of zinc increased at 24 h, preceding the increase of taurine. The axonal transport of [3H]taurine from the optic tectum to the retina was not affected in fish with or without crush of the optic nerve at early periods after the injection, indicating an increase of it post-lesion. Zinc sulfate produced a bell-shaped concentration dependency on in vitro outgrowth, with stimulation at 0.05 microM, and inhibition at higher levels, also increased the effect of 4 mM taurine at 0.02 microM, but diminished it at higher concentrations in the medium. TPEN decreased outgrowth at 1 nM, but not at 0.5 nM, although the simultaneous presence of 4 mM taurine and 0.5 nM TPEN decreased outgrowth respecting the stimulation by taurine alone. The intraocular administration of TPEN decreased outgrowth in vitro, an effect counteracted by the addition of 4 mM taurine to the culture medium. DTPA decreased outgrowth from 10 microM in the medium. The present results indicate that an optimal zinc concentration is necessary for outgrowth of goldfish retinal explants and that, in zinc deficient retina, taurine could stimulate outgrowth. In addition, the observations of variations in tissue concentrations and of the effects of intraocular administration of TPEN indicate that these effects could occur in vivo. PMID- 16362768 TI - Semax, an ACTH(4-10) analogue with nootropic properties, activates dopaminergic and serotoninergic brain systems in rodents. AB - Corticotrophin (ACTH) and its analogues, particularly Semax (Met-Glu-His-Phe-Pro Gly-Pro), demonstrate nootropic activity. Close functional and anatomical links have been established between melanocortinergic and monoaminergic brain systems. The aim of present work was to investigate the effects of Semax on neurochemical parameters of dopaminergic- and serotonergic systems in rodents. The tissue content of 5-hydroxyindoleacetic acid (5-HIAA) in the striatum was significantly increased (+25%) 2 h after Semax administration. The extracellular striatal level of 5-HIAA gradually increased up to 180% within 1-4 h after Semax (0.15 mg/kg, ip) administration. This peptide alone failed to alter the tissue and extracellular concentrations of dopamine and its metabolites. Semax injected 20 min prior D: -amphetamine dramatically enhanced the effects of the latter on the extracellular level of dopamine and on the locomotor activity of animals. Our results reveal the positive modulatory effect of Semax on the striatal serotonergic system and the ability of Semax to enhance both the striatal release of dopamine and locomotor behavior elicited by D-amphetamine. PMID- 16362769 TI - Problems associated with biological markers of Alzheimer's disease. AB - The etiopathogenesis of Alzheimer's disease (AD) is still unclear, although clinical diagnostic criteria exist and the neuropathology of AD has been studied in great detail during the last 20 years. The present study addresses certain problems in the search for biological markers for the diagnosis, as well as in the follow-up of the course of AD and its differential diagnosis and reports some of our own observations in comparison with other studies. These include protein, genetic and neuroimaging markers. The definitions of biological markers and search strategies are also discussed. PMID- 16362770 TI - Effects on cholinergic markers in rat brain and blood after short and prolonged administration of donepezil. AB - Donepezil is a selective inhibitor of acetylcholinesterase (AChE) clinically used for treating Alzheimer's disease. Cholinergic effects after short-term exposure of donepezil (up to 12 h) have been extensively studied in rats, but few have addressed the potential long-term effects. After 14 days administration (1x3 mg/kg, decapitation 4 h after the last injection) the cerebral acetylcholine level was increased by 35% and the AChE activity was decreased by 66% and 32% in brain and blood, respectively. No change was detected in choline acetyltransferase activity, or the levels of vesicular acetylcholine transporter, choline transporter, or muscarinic receptors. Expression of various cholinergic genes was unaffected. Preliminary results of AChE activity in human blood showed 60-97% and 43-89% of pre-exposed level after one and three days of donepezil administration (5 mg daily), respectively. In conclusion, donepezil exposure in rats at doses that do not inhibit brain AChE continuously during the day, will not lead to tolerance development. PMID- 16362771 TI - Organotypic hippocampal slice cultures: a model system to study basic cellular and molecular mechanisms of neuronal cell death, neuroprotection, and synaptic plasticity. AB - The hippocampus has become one of the most extensively studied areas of the mammalian brain, and its proper function is of utmost importance, particularly for learning and memory. The hippocampus is the most susceptible brain region for damage, and its impaired function has been documented in many human brain diseases, e.g. hypoxia, ischemia, and epilepsy regardless of the age of the affected patients. In addition to experimental in vivo models of these disorders, the investigation of basic anatomical, physiological, and molecular aspects requires an adequate experimental in vitro model, which should meet the requirements for well-preserved representation of various cell types, and functional information processing properties in the hippocampus. In this review, the characteristics of organotypic hippocampal slice cultures (OHCs) together with the main differences between the in vivo and in vitro preparations are first briefly outlined. Thereafter, the use of OHCs in studies focusing on neuron cell death and synaptic plasticity is discussed. PMID- 16362772 TI - Epileptogenesis after experimental focal cerebral ischemia. AB - Cerebrovascular diseases are one of the most common causes of epilepsy in adults, and the incidence of stroke-induced epileptogenesis is increasing as the population ages. The mechanisms that lead to stroke-induced epileptogenesis in a subpopulation of patients, however, are still poorly understood. Recent advances in inducing epileptogenesis in rodent focal ischemia models have provided tools that can be used to identify the risk factors and neurobiologic changes leading to development of epilepsy after stroke. Here we summarize data from models in which epileptogenesis has been studied after focal ischemia; photothrombosis, middle cerebral artery (MCA) occlusion with filament, and endothelin-1-induced MCA occlusion. Analysis of the data indicates that neurobiologic changes occurring during stroke-induced epileptogenesis share some similarities to those induced by status epilepticus or traumatic brain injury. PMID- 16362773 TI - Mechanisms and significance of the increased brain uptake of tryptophan. AB - Changes in brain tryptophan concentrations may affect the synthesis of brain serotonin (5-hydroxytryptamine, 5-HT). Concentrations of tryptophan are regulated more than those of any other amino acid. Such stimuli as acute stress, carbohydrate ingestion, and treatment with various drugs increase the brain content of tryptophan. Treatment of rats and mice with interleukin-1 (IL-1), interleukin-6 (IL-6), lipopolysaccharide (LPS), and beta-adrenoceptor agonists, as well as a variety of stressors, such as footshock and restraint, all increase brain concentrations of tryptophan. The peak effect following both acute stress and beta-adrenoceptor agonist administration occurs within 30-60 min, whereas the peak effect following LPS and the cytokines occurs much later at around 4-8 h. Experiments using the ganglionic blocker chlorisondamine, and beta-adrenoceptor antagonists suggest that the sympathetic nervous system plays an important role in the modulation of brain tryptophan concentrations. The mechanisms involved in the increases observed in brain tryptophan are discussed, as well as their possible biological significance. PMID- 16362774 TI - Characteristics of GABA release in mouse brain stem slices under normal and ischemic conditions. AB - GABA is known to be the inhibitory neurotransmitter in the majority of brain stem nuclei. The release of GABA has been extensively studied both in vivo and in vitro in higher brain areas, whereas the mechanisms of release in the brain stem have not been systemically characterized. The properties of preloaded [3H]GABA were now investigated in mouse brain stem slices, using a superfusion system. The basal release was enhanced by K+ stimulation (50 mM K+) and under various cell damaging conditions (ischemia, hypoglycemia, the presence of free radicals and metabolic poisons). No K+-stimulated release was discernible in the absence of Ca2+, indicating that the release was at least partly Ca2+-dependent. Moreover, the release was increased when Na+ or Cl- was omitted from the superfusion medium. GABA and beta-alanine stimulated the release, confirming the involvement of the reversed function of GABA transporters. Incubation of the slices with the anion channel inhibitors diisothiocyanostilbene and 4-acetamido-4' isothiocyanostilbene-2,2'-disulphonate and with the Cl- uptake inhibitor 9 anthracenecarboxylic acid also reduced GABA release, demonstrating that a part of it comprises leakage through anion channels. All these mechanisms were involved in the ischemia-induced GABA release, which was over 4-fold greater than the release in normoxia. Contrary to the other brain areas, GABA release in the brain stem was not affected by ionotropic glutamate receptors but may be modulated by metabotropic receptors. This ischemia-induced GABA release might constitute an important mechanism against excitotoxicity, protecting the brain stem under cell damaging conditions. PMID- 16362775 TI - Effects of retigabine on the neurodegeneration and extracellular glutamate changes induced by 4-aminopyridine in rat hippocampus in vivo. AB - We have previously shown that microdialysis perfusion of the K+ channel blocker 4 aminopyridine (4-AP) in rat hippocampus induces convulsions and neurodegeneration, due to the stimulation of glutamate release from synaptic terminals. Retigabine is an opener of the KCNQ2/Q3-type K+ channel that possesses antiepileptic action and may be neuroprotective, and we have therefore studied its effect on the hyperexcitation, the neuronal damage and the changes in extracellular glutamate induced by 4-AP. Retigabine and 4-AP were co-administered by microdialysis in the hippocampus of anesthetized rats, with simultaneous recording of the EEG, and the extracellular concentration of glutamate was measured in the microdialysis fractions. In 70-80% of the rats tested retigabine reduced the 4-AP-induced stimulation of glutamate release and prevented the neuronal damage observed at 24 h in the CA1 hippocampal region. However, retigabine did not block the EEG epileptic discharges and their duration was reduced in only 20-25% of the tested animals. We conclude that the neuroprotective action of retigabine is probably due to the blockade of the 4-AP induced stimulation of glutamate release. This inhibition, however, was not sufficient to block the epileptic activity. PMID- 16362776 TI - Characteristics and regulation of glycine transport in Bergmann glia. AB - In the vertebrate CNS, glycine acts as an inhibitory neurotransmitter and as the obligatory coagonist of glutamate at N-methyl-D-aspartate receptors. These roles depend on extracellular glycine levels, regulated by Na+/Cl--dependent transporters GLYT1, present mainly in glial cells, and GLYT2, predominantly neuronal. In Bergmann glia, GLYT1 mediates both, glycine uptake and efflux, which, in turn, influences excitatory neurotransmission at Purkinje cell synapses. The biochemical properties of GLYTs and their regulation by signaling pathways in these cells are largely unknown. We characterized Gly uptake in confluent primary cultures of Bergmann glia from chick cerebellum. Transport was found to be energy- and Na+-dependent, and was resolved into a high (Km=25 microM) and a low affinity (Km=1.1 mM) components identified as GLYT1 and transport System A, respectively. Results show that high affinity transport by GLYT1 is regulated by calcium from intracellular stores, calmodulin, and myosin light chain kinase through an actin cytoskeleton-mediated action. PMID- 16362778 TI - Epidermal growth factor receptor is a common element in the signaling pathways activated by cell volume changes in isosmotic, hyposmotic or hyperosmotic conditions. AB - Changes in external osmolarity, including both hyper- or hyposmotic conditions, elicit the tyrosine phosphorylation of a number of tyrosine kinase receptors (TKR). We show here that the epidermal growth factor receptor (EGFR) is activated by both cell swelling (hyposmolarity, isosmotic urea, hyperosmotic sorbitol) or shrinkage (hyperosmotic NaCl or raffinose) and discuss the mechanisms by which these apparently opposed conditions come to the same effect, i.e., EGFR activation. Evidence suggests that this results from early activation of integrins, p38 and tyrosine kinases of the Src family, which are all activated in the two anisosmotic conditions. TKR transactivation by integrins and p38 is likely occurring via an effect on the metalloproteinases. Information discussed in this review, points to TKR as elements in osmotransduction as a useful mechanism to amplify and diversify the initial response to anisosmolarity and cell volume changes, due to their privileged situation as convergence point for numerous intracellular signaling pathways. The variety of effector pathways connected to TKR is advantageous for the cell to cope with the changes in cell volume including adaptation to stress, cytoskeleton remodeling, adhesion reactions, cell survival and the adaptive mechanisms to ultimately restore the original cell volume. PMID- 16362777 TI - Plasticity of GABAA receptors in brains of rats treated with chronic intermittent ethanol. AB - The study of alcohol dependence mechanisms has been aided by work in rodents, where regimens of intermittent chronic administration with repeated episodes of intoxication and withdrawal can be coupled with controlled timing of in vitro studies and the possibility of relating them to behavior. The chronic intermittent ethanol (CIE) model in the rat has been found to be a good model of human alcohol dependence, showing persistent signs of withdrawal and self administration. Studies in CIE rats suggest that plastic changes in GABA-mediated inhibition involving the GABAA receptor system may be responsible for the behavioral alterations. Here we summarize a combination of evidence that the alcoholic rat CIE model demonstrates changes in GABAA receptor subunit levels, in receptor localization, and in physiology and pharmacology, leading to alterations in behavior that contribute to the hyperexcitable alcohol withdrawal state (anxiety, insomnia, seizure susceptibility) and alcohol dependence. PMID- 16362779 TI - Alterations in the development of rat cerebellum and impaired behavior of juvenile rats after neonatal 6-OHDA treatment. AB - The effects of neonatal systemic administration of the neurotoxin 6 hydroxydopamine (6-OHDA) on cerebellum development and behavior were studied in juvenile rats. The methods employed were immunohistochemistry, in situ hybridization, ligand binding, and behavioral testing. The results revealed, for the first time, that 6-OHDA treatment alters Bergmann glial cells and reduced the expression GABAA receptor subtypes alpha1 and alpha6 especially in granule cells. The Bergmann glial cells were abnormally located and structurally different (e.g., no intimate associations with Purkinje cells). Significant microglial activation was also observed. The animals showed impairment in behavior, especially in their orientation to a novel environment. Recent data on neuron glia interactions support the conclusion that the observed structural changes in Bergmann glia and granular neurons disrupted the normal functioning of the Purkinje cells which then in turn resulted in the impaired sensory-motor coordination at least in juvenile rats. This paper is a summary of previously published work and some recent data in this field obtained at our laboratory. PMID- 16362780 TI - Effect of moxibustion on dopaminergic and serotonergic systems of rat nucleus accumbens. AB - Acupuncture and moxibustion are traditional medical treatments that have come to play important roles in complementary and alternative medicines. Moxibustion also has a long history as a folk remedy in Japan, particularly due to the technical simplicity and selective efficacy on certain types of disease and distress. This study examined the effects of moxibustion focusing on the brain reward system, particularly in the nucleus accumbens. The effects of moxibustion stimulation at various sites and frequencies on monoamine levels of adult male Sprague-Dawley rats were examined using high-preformance liquid chromatography of dissected nucleus accumbens tissues. The rats weighing 290-310 g were divided into 3 groups according to the moxibustion point used: hindlimb, lumbar or parietal points. Each group was further divided into 3 subgroups, with stimulation for 10 consecutive days, for 1 day, or sham treatment (control). On each day of stimulation, 5 moxibustion cones with a peak temperature of 200 degrees C were applied consecutively. Stimulation of any point on 1 day only did not change dopamine or serotonin levels, but lumbar stimulation significantly increased the metabolic turnover of dopamine. Conversely, stimulation for 10 consecutive days resulted in significantly decreased serotonin levels for hindlimb and parietal stimulations, and significantly increased 5-hydroxyindolacetic acid/serotonin ratio for hindlimb stimulation. These results suggest that the metabolic turnover of serotonin release may be accentuated by moxibustion in a reward-related brain area. Moxibustion over consecutive days, especially that to peripheral regions, appears most efficient to influence on monoamine levels in the nucleus accumbens. PMID- 16362781 TI - Taurine interaction with neurotransmitter receptors in the CNS: an update. AB - Taurine appears to have multiple functions in the brain participating both in volume regulation and neurotransmission. In the latter context it may exert its actions by serving as an agonist at receptors of the GABAergic and glycinergic neurotransmitter systems. Its interaction with GABAA and GABAB receptors as well as with glycine receptors is reviewed and the physiological relevance of such interactions is evaluated. The question as to whether local extracellular concentrations of taurine are likely to reach the threshold level for the pertinent receptor populations cannot presently be answered satisfactorily. Hence more sophisticated analytical methods are warranted in order to obtain a definite answer to this important question. PMID- 16362784 TI - Restricted-expressed proliferation-associated protein (Repp86) expression in squamous cell carcinoma of the oral cavity. AB - PURPOSE: To determine the expression of repp86 (restricted-expressed protein of 86 kDa theoretical molecular mass), a proliferation- associated protein expressed in S-, G(2)- and M-phases of the cell cycle, in samples of normal mucosa as well as squamous cell carcinoma of the oral cavity (OSCC). PATIENTS AND METHODS: The repp86 labeling index (LI) was determined imunohistochemically in ten samples of normal oral mucosa and 59 samples of OSCC. Repp86 LI was correlated with tumor stage, histopathologic grading, and the expression of Ki-67 and topoisomerase IIalpha. RESULTS: Repp86 was detectable in all tissues analyzed. The mean LI was 4.7% for normal mucosa and 18.4% for squamous cell carcinoma (p < 0.0001). Repp86 expression was not related to tumor size, lymph node invasion, or histopathologic grading but was positively correlated with Ki-67 index (r = 0.48; p < 0.01) as well as with topoisomerase IIalpha (r = 0.39; p < 0.01). Ki-67 and topoisomerase IIalpha levels were also significantly correlated with each other (r = 0.34; p < 0.05). CONCLUSION: These results indicate that repp86 expression can be an additional proliferation marker among Ki-67 and topoisomerase IIalpha in OSCC. Further research will be directed at the evaluation of the prognostic value of repp86 expression in OSCC as well as in leukoplakia and early-stage OSCC. PMID- 16362785 TI - Role of interstitial PDR brachytherapy in the treatment of oral and oropharyngeal cancer. A single-institute experience of 236 patients. AB - PURPOSE: To evaluate the role of pulsed-dose-rate interstitial brachytherapy (PDR IBT) in patients with head-and-neck malignancies. PATIENTS AND METHODS: From October 1997 to December 2003, 236 patients underwent PDR IBT for head-and-neck cancer at the authors' department. 192 patients received brachytherapy as part of their curative treatment regimen after minimal non-mutilating surgery, 44 patients were treated with irradiation alone. 144 patients had sole IBT (median D(REF) = 56 Gy), in 92 patients IBT procedures (median D(REF) = 24 Gy) were performed in combination with external irradiation. The pulses (0.4-0.7 Gy/h) were delivered 24 h a day with a time interval of 1 h between two pulses. The analysis of tumor control, survival and treatment-related toxicity was performed after a median follow-up of 26 months (6-75 months). RESULTS: At the time of analysis permanent local tumor control was registered in 208 of 236 patients (88%). At 5 years overall survival and local recurrence-free survival of the entire group were 82-73% and 93-83% for T1/2, and 56% and 83% for T3/4, respectively. Soft-tissue necrosis was seen in 23/236 patients (9.7%) and bone necrosis in 17/236 patients (7.2%). No other serious side effects were observed. CONCLUSION: PDR IBT with 0.4-0.7 Gy/h and 1 h between pulses is safe and effective. These results confirm that PDR IBT of head-and-neck cancer is comparable with low-dose-rate (LDR) brachytherapy - equally effective and less toxic. PMID- 16362786 TI - Reirradiation of recurrent WHO grade III astrocytomas using fractionated stereotactic radiotherapy (FSRT). AB - PURPOSE: To assess the effect of reirradiation in recurrent WHO grade III astrocytomas. PATIENTS AND METHODS: From January 1995 to July 2003, 40 patients with grade III gliomas were treated with fractionated stereotactic reirradiation at the time point of recurrence. Median size of planning target volume for reirradiation was 56.2 ml (range 25.1-296.2 ml). A median target total dose of 36 Gy (range 20-57.6 Gy) was applied using a median fractionation of 5 x 2 Gy/week with a 6-MeV linear accelerator. RESULTS: Radiotherapy was well tolerated by all patients. No toxicities > CTC grade 2 developed. Median overall survival calculated from the time point of primary diagnosis was 48 months (range 7-180 months). The 5- and 10-year overall survival rates were 49.5% and 24.7%, respectively. From the time point of reirradiation, median survival was 16 months (range 1-98 months). Median progression-free survival from the time point of reirradiation was 8 months (range 1-72 months). No prognosticators for survival or progression-free survival after reirradiation could be identified. CONCLUSION: Fractionated stereotactic radiotherapy is well tolerated and effective in patients with recurrent grade III astrocytomas. PMID- 16362787 TI - Neutron activation of patients following boron neutron capture therapy of brain tumors at the high flux reactor (HFR) Petten (EORTC Trials 11961 and 11011). AB - BACKGROUND AND PURPOSE: At the High Flux Reactor (HFR), Petten, The Netherlands, EORTC clinical trials of Boron Neutron Capture Therapy (BNCT) have been in progress since 1997. BNCT involves the irradiation of cancer patients by a beam of neutrons, with an energy range of predominantly 1 eV to 10 keV. The patient is infused with a tumor-seeking, (10)B-loaded compound prior to irradiation. Neutron capture in the (10)B atoms results in a high local radiation dose to the tumor cells, whilst sparing the healthy tissue. Neutron capture, however, also occurs in other atoms naturally present in tissue, sometimes resulting in radionuclides that will be present after treatment. The patient is therefore, following BNCT, radioactive. The importance of this induced activity with respect to the absorbed dose in the patient as well as to the radiation exposure of the staff has been investigated. MATERIAL AND METHODS: As a standard radiation protection procedure, the ambient dose equivalent rate was measured on all patients following BNCT using a dose ratemeter. Furthermore, some of the patients underwent measurements using a gamma-ray spectrometer to identify which elements and confirm which isotopes are activated. RESULTS: Peak levels, i.e., at contact and directly after irradiation, are of the order of 40-60 muSv/h, falling to < 10 muSv/h 30-50 min after treatment. The average ambient dose equivalent in the first 2 h at a distance of 2 m from the patient is in the order of 2.5 muSv. The ambient dose equivalent rate in 2 m distance from the patient's head at the earliest time of leaving the reactor center (20 min after the end of treatment) is far less than 1 muSv/h. The main radioisotopes were identified as (38)Cl, (49)Ca, and (24)Na. Furthermore, in two patients, the isotopes (198)Au and (116m)In were also present. The initial activity is predominantly due to (49)Ca, whilst the remaining activity is predominantly due to (24)Na. CONCLUSION: The absorbed dose resulting from the activated isotopes in the irradiated volume is in the order of < 1% of the prescribed dose and therefore does not add a significant contribution to the absorbed dose in the target volume. In other parts of the patient's body, the absorbed dose by induced activity is magnitudes smaller and can be neglected. The levels of radiation received by staff members and non-radiation workers (i.e., accompanying persons) are well below the recommended limits. PMID- 16362788 TI - Proton or stereotactic photon irradiation for posterior uveal melanoma? A planning intercomparison. AB - BACKGROUND AND PURPOSE: Proton and stereotactic radiotherapy with photons (SRT) are both used to treat choroidal melanomas in proximity to optic disk and fovea centralis, a situation where plaque therapy is prone to complications. A comparative treatment- planning study was done to assess the capability of both modalities to preserve vision. PATIENTS AND METHODS: In ten patients treated with 68-MeV protons, SRT with 6-MV photons was planned. Structures most important for visual acuity (fovea and optic disk, optic nerve) were contoured identically for both therapies. Safety margins of 1.5 mm for proton therapy were reduced to 1.0 mm for SRT. RESULTS: Proton-beam therapy was superior in eight of ten situations, and this result did not differ significantly by changes in the weighting of the different parameters analyzed. CONCLUSION: When dose deposition to those structures most important for the preservation of vision is taken into account, under the conditions examined proton therapy offers an advantage in the majority of the patients evaluated. PMID- 16362789 TI - A prospective three-dimensional analysis about the impact of differences in the clinical target volume in prostate cancer irradiation on normal-tissue exposure. A potential for increasing the benefit/risk ratio. AB - BACKGROUND AND PURPOSE: Rectal toxicity following external-beam irradiation of prostate cancer correlates with the exposed percentage of rectal volume. Recently, it has been recommended to reduce the volume of the seminal vesicles that should be included in the clinical target volume (CTV). The purpose of this study was to quantitatively assess the impact of this CTV reduction on the expected rectal and bladder dose sparing. PATIENTS AND METHODS: 14 patients with localized prostate cancer undergoing external-beam radiotherapy were investigated. The prostate, the prostate + entire seminal vesicles, or the prostate + proximal seminal vesicles were delineated as CTV. Treatment plans were generated and compared concerning rectum and bladder dose-volume histograms (DVHs). RESULTS: The exposure of rectum and bladder volume was significantly lower in case of irradiation of the prostate only compared to inclusion of the proximal or entire seminal vesicles into the CTV. The reduction of the CTV from prostate + entire seminal vesicles to prostate + proximal seminal vesicles led to a significant reduction of the rectal and bladder dose exposure. CONCLUSION: Reduction of the CTV to the prostate only, or to the prostate + proximal seminal vesicles led to significant rectal and bladder dose sparing compared to irradiation of the prostate + entire seminal vesicles. In patients with a higher risk for seminal vesicles involvement, irradiation of the prostate + proximal seminal vesicles should be preferred. In case of a need for irradiation of the entire seminal vesicles, patients should be informed about a higher risk for chronic rectal toxicity and, possibly, for bladder complications. PMID- 16362790 TI - Dose delivery accuracy of therapeutic photon and electron beams at low monitor unit settings. AB - PURPOSE: Dose delivery accuracy at low monitor units (LMU) was evaluated for photon and electron beams. Knowledge of this study is required for few dosimetric applications and to know the dose delivered to the patient when the treatment is delivered with few monitor units (MU). MATERIAL AND METHODS: Dose measurements were carried out for photon and electron beams with 0.6 cm(3) PTW ion chamber in white polystyrene phantom at D(max) with a field size of 10 x 10 cm(2) at 100 cm FSD. The relative dose, which is the ratio of dose delivered per MU at the testing to that of the calibration condition, was found out. RESULTS: Significant deviation (+20% to +25%) in dose delivery was noticed for photon and electron beams (+39% to +45%) at LMU settings. Slightly higher inaccuracy in dose delivery was noticed for 6-MV compared to 18-MV photons. The deviation in dose delivery for electron beams was found to be energy-independent and the pattern of variation was similar for all electron energies. CONCLUSION: The dose delivery accuracy at LMU settings has to be ascertained before implementing conformal and IMRT (intensity- modulated radiotherapy) techniques. When there is dose nonlinearity, the treatment delivered with multiple small MU settings can result in significant error in dose delivery. PMID- 16362791 TI - [The prognostic value of time parameters in adjuvant radiotherapy of head and neck cancer. A retrospective analysis of 138 patients]. AB - PURPOSE: To answer the question, how the parameters waiting time, radiation treatment time and overall treatment time (OTT) influenced the endpoints overall (OS), event-free (EFS) and local recurrence-free survival (LRFS) in patients with locally advanced head-and-neck cancer, who had received postoperative radiotherapy. PATIENTS AND METHODS: 138 patients were included into a retrospective analysis from 10/1993 to 05/2000. Besides the time parameters waiting time, radiation treatment time and OTT, tumor- and therapy-related parameters (T-, N-, R-status, grading, tumor site, surgical technique, and postoperative hemoglobin < 12 g/dl) with potential impact on the endpoints were investigated in the univariate analysis (Kaplan-Meier log-rank test). Individual parameters with a significant impact (p = 0.05) were subjected to a multivariate Cox regression analysis. RESULTS: Besides a postoperative hemoglobin value < 12 g/dl, in the univariate analysis an OTT >/= 105 days negatively influenced all endpoints, as well as a radiation treatment time >/= 60 days. On multivariate Cox regression analysis, postoperative hemoglobin < 12 g/dl and an OTT >/= 105 days were identified as independent negative prognostic factors for all endpoints. CONCLUSION: The waiting time should be managed according to the ASARA (as short as reasonably achievable) recommendation, radiation treatment should not be protracted exceeding an overall treatment of 105 days. Generally, time parameters should be routinely included in the standard tumor documentation, thus facilitating further evaluation of these prognostically relevant factors. PMID- 16362793 TI - Immunomodulation of autoimmune and inflammatory diseases with intravenous immunoglobulin. AB - Intravenous immunoglobulin (IVIg) has been used in the treatment of primary and secondary antibody deficiencies for over two decades. Since the early 1980s, the therapeutic efficacy of IVIg has been established in idiopathic thrombocytopenic purpura, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, myasthenia gravis, dermatomyositis and Kawasaki syndrome, and the prevention of graft versus host disease in recipients of allogeneic bone marrow transplants. Its use has also been reported in a large number of other autoimmune and systemic inflammatory conditions. In this review, we discuss the mechanisms by which IVIg exerts immunomodulatory effects in immune pathologies. PMID- 16362794 TI - Biological therapy with monoclonal antibodies: a novel treatment approach to autoimmune disease. AB - Most autoimmune diseases (ADs) are still associated with high morbidity and mortality despite the use of a wide range of drugs that can delay their progression, control their symptoms, but never bring about a complete cure. This failure has aroused interest in new forms of monoclonal antibody-based experimental immunotherapy (IT), aiming at targeting cellular antigens or cytokines involved in the pathogenesis of ADs. The first part of this review offers a general overview of the molecular mechanisms that mediate the immune response and the molecule regarded as potential IT targets. A critical evaluation will then be made of some forms of IT, with particular emphasis on TNF-alpha and CD20-blocking reagents. Lastly an account will be given of active IT whereby an endogenous response against antigens regarded as the target of passive IT can be induced by anti-idiotype or peptides. PMID- 16362795 TI - Polymorphisms of the TP53 codon 72 and WRN codon 1367 in individuals from Northern Brazil with gastric adenocarcinoma. AB - Gastric cancer is the second most frequent type of neoplasia and also the second most common cause of death in the world. TP53 codon 72, which produces variant proteins with an arginine (Arg) or proline (Pro), has been reported to be associated with cancers of the lung, oesophagus, stomach and cervix. Werner's syndrome (WS) is a premature ageing disease caused by a mutation in the WRN gene. The WRN protein acts as a DNA helicase and as an exonuclease. WRN codon 1367 produces variant proteins with an Arg or cysteine (Cys). This polymorphism has been studied, in order to understand the clinical impact of the molecular variants in WS and in age-related disorders. In the present study, the TP53 codon 72 and the WRN codon 1367 polymorphisms were investigated in 54 gastric adenocarcinoma patients (31 diffuse-type and 25 intestinal-type) and 54 controls. DNA samples were extracted, and PCR-RFLP was utilised for genotyping TP53 codon 72 and WRN codon 1367. The allele frequencies of the TP53 polymorphism were: Arg=0.74 and Pro=0.26. The allele frequencies of the WRN polymorphism were: Cys=0.73 and Arg=0.27. The crude genotypic frequencies in gastric cancer patients were similar to those of the controls, but in the WRN codon 1367 polymorphisms the mean age tended to be higher in the Arg/Arg genotypes. There also was an association, although not statistically significant, between the presence of Helicobacter pylori and the genotypes Cys/Cys and Cys/Arg and a higher percentage of cardia cancer among the Arg/Arg genotypes, and of non-cardia cancer among genotypes Cys/Cys and Cys/Arg. These findings may be a reflection of differences in the interaction between WRN codon 1367 polymorphisms and local factors in the stomach. To our knowledge, this is the first study to examine a genetic polymorphism of the WRN gene in cancer. The precise mechanisms of action of the TP53 and WRN polymorphisms involved in the aetiopathogeny of this disease need further investigation. PMID- 16362796 TI - Association of interleukin-1B and interleukin-1RN polymorphisms with gastric cancer in a high-risk population of Costa Rica. AB - Several risk factors have been associated with gastric cancer, among them Helicobacter pylori infection. This bacterium yields inflammation, the degree of which depends on the bacterial strain and the severity of the host response. The inflammatory response involves a complex cytokine network. Recently, polymorphisms of the genes coding for interleukin-1beta (IL-1B), interleukin-1Ra (ILRN) and interleukin-10 have been associated with an increased risk of gastric cancer. In order to determine the association of the IL-1B, IL-1RN and IL-10 polymorphisms with gastric cancer in a high-risk Costa Rican population, we analysed purified DNA of 58 gastric cancer patients, 99 controls and 41 patients classified as group I or II, according to the Japanese classification. Genotyping was carried out by PCR, PCR-RFLP and pyrosequencing analysis. We did not find any association of the IL-1B-31, IL-1B-511 and IL-10 polymorphisms with the risk for developing gastric cancer in the studied population. Carriers of the IL-1B+3954T/ had an increased risk for developing gastric cancer (OR 3.7; 95%CI: 1.34-10.2). Also we found an increased risk for developing gastric cancer for allele 2 heterozygotes of the IL-1RN (OR 2.94; 95%CI: 1.09-7.93). This is the first time that IL-1B+3954 has been associated with gastric cancer. This is one of the first studies trying to describe the role played by IL-1B, IL-1RN and IL-10 genetic polymorphisms in gastric cancer in one of the highest risk American countries. Further investigation on American countries is needed. PMID- 16362797 TI - Influence of immunisation with Mycobacterium bovis bacillus Calmette-Guerin on the sensitisation to inhaled allergens after infection with respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) may play an important role in allergic diathesis by creating a Th2-type immune response. Mycobacterium bovis bacillus Calmette-Guerin (BCG) is known to induce a Th1-type immune response, but the association of BCG vaccination and the suppression of allergy development remain controversial. We investigated the influence of BCG vaccination on the immune response to RSV in a mouse model. Balb/c mice were BCG vaccinated, RSV infected and ovalbumin (OVA) challenged. Mice were sacrificed one, two and four weeks after allergen exposure. Bronchoalveolar lavage was performed. Alveolar macrophages and lymphocytes from spleens and lung-associated lymph nodes were investigated for cytokine production and cell proliferation. Serum was tested for allergen-specific immunoglobulin-E (IgE). Lung eosinophilia was diminished by BCG immunisation. OVA-specific serum IgE was increased regardless of prior BCG vaccination. Interleukin-4 secretion of spleen lymphocytes increased in BCG vaccinated mice only one week after allergen exposure but was comparable to non vaccinated mice at four weeks. The reactivity of spleen lymphocytes towards concanavalin-A to secrete interferon-gamma was increased in the vaccinated group at the end of the observation period. Interleukin-6 and tumour necrosis factor alpha secretion of alveolar macrophages as well as proliferation of stimulated thoracic lymph node cells were increased and prolonged in vaccinated mice. BCG immunisation led to a local suppression of the allergic reaction within the lung. No reduction of systemic IgE production was observed. Further studies are necessary to determine a possible time dependence of BCG immunisation. PMID- 16362798 TI - NCL-CD30 staining of epithelial cells in the basal germinative layer of the epidermis and epithelial buds during foetal skin development. AB - The fact that the CD30 molecule can mediate signals for cell proliferation or apoptosis prompted us to perform a systematic investigation of CD30 antigen expression in embryonal tissues during proliferation and differentiation stages. We first targeted the foetal human intestinal cryptae cells with positive results. The epidermis is a dynamic epithelium that is constantly renewed throughout life. The basal layer, attached to the basement membrane, contains the dividing cells of the skin and as cells move up from this layer they undergo differentiation, ending in the formation of a terminally differentiated anucleate cell called squame. It is intriguing to find out if cells in the basal layer can express the CD30 antigen. We investigated the immunohistochemical expression of CD30 antigen in 15 paraffin-embedded tissue samples representing epidermis and epidermal buds from foetuses after spontaneous abortion in the 8th, 10th and 12th weeks of gestation, respectively, using the monoclonal antibody NCL-CD30. A Northern blotting analysis was additionally performed. The results showed that: (1) the epithelial cells of the epidermis in the developing skin express the CD30 antigen; (2) CD30 expression in these epithelial cells is higher in cases of hormonal administration than in normal gestation; (3) a similar positive reaction involved the epidermal buds associated with the development of the skin appendages. Northern blots of tissue sections using a CD30 cDNA probe detected mRNAs of the same molecular mass and variety similarly to those in the positive control cell line HUT 102. PMID- 16362799 TI - Changes in serum and red blood cell membrane lipids in patients treated with interferon ribavirin for chronic hepatitis C. AB - One of the side effects by interferon ribavirin (I/R) treatment is haemolytic anemia, causing some patients to discontinue I/R treatment. The exact mechanism of I/R-induced anemia is unknown. The aim of this study is to evaluate the effects of I/R treatment on the serum lipid and red blood cell (RBC) membrane lipid profiles of patients with chronic hepatitis C (CHC) and the association between changes of RBC membrane lipids and haemolytic anemia by I/R treatment. Fourteen patients with CHC were treated with I/R and their serum lipid profiles were studied. In addition, in seven of the 14 patients, the RBC membrane lipid profiles were analysed. In the RBC membrane lipid composition, the total cholesterol, total phospholipids and cholesterol/phospholipids (C/PL) ratio were significantly increased. Phosphatidylcholine (PC) and the phosphatidylcholine/ sphingomyelin (PC/SM) ratio were significantly decreased and other phospholipid fractions were significantly increased. Changes in the serum lipids and RBC membrane lipid profiles of patients with CHC treated with I/R were shown. Especially, a decrease in the RBC deformability and membrane fluidity by changes in these RBC membrane lipids was supposed and it is suggested that those changes may result in haemolytic anemia by I/R treatment. PMID- 16362800 TI - Apolipoprotein E polymorphism is not associated with lipid levels and coronary artery disease in Greek patients with familial hypercholesterolaemia. AB - Familial hypercholesterolaemia is a genetic disorder characterised by high low density lipoprotein (LDL) cholesterol concentrations, which frequently gives rise to premature coronary artery disease (CAD). The clinical expression of familial hypercholesterolaemia is highly variable even in patients carrying the same LDL receptor gene mutation. This variability may be due to environmental and other genetic factors. Apolipoprotein E (Apo-E) has been extensively studied for its effects on the phenotype of familial hypercholesterolaemia. In this study we examined the influence of Apo-E genotype on lipid parameters and the incidence of CAD in 93 Greek patients with familial hypercholesterolaemia. Apo-E E2, E3 and E4 allele frequencies were 0.06, 0.86 and 0.09 respectively. The levels of total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, apolipoproteins A and B and lipoprotein alpha did not differ significantly among carriers and non carriers of the E4 allele. The prevalence of CAD and hypertension did not differ either. Our results suggest that the E4 allele is not associated with lipid levels or with the prevalence of CAD among familial hypercholesterolaemia patients of the Greek population. PMID- 16362801 TI - Transanal endoscopic microsurgical resection of pT1 rectal tumors. AB - PURPOSE: Transanal endoscopic microsurgery has emerged as an improved method of transanal excision of neoplasms because its enhanced visibility, superior optics, and longer reach permit a more complete excision and precise closure. This study will show that transanal endoscopic microsurgical treatment of pT1 rectal cancers is safe and achieves low local recurrence and high survival rates. METHODS: Retrospective review performed of all pT1 rectal cancers treated by a single surgeon (TS) using transanal endoscopic microsurgery between 1991 and 2003. Patient age, gender, tumor distance from the anal verge, lesion size, operative time, blood loss, complications, recurrence, and survival rates were prospectively recorded. RESULTS: Fifty-three patients (average age, 65.6 (range, 31-89) years) were studied. Forty-nine percent were male. Average tumor distance from the anal verge was 7 (range, 0-13) cm; average size was 2.4 (range, 1-10) cm. Radiation and/or chemotherapy were not administered. Sixteen patients had pT1 lesions removed piecemeal during colonoscopy; there was no residual tumor after transanal endoscopic microsurgical resection of the polyp site. Mean follow-up was 2.84 years. Fifty-one percent had longer than two-year follow-up. For the entire group, there were four recurrences (7.5 percent) occurring at 9 months, 15 months, 16 months, and 11 years. Two were treated with abdominoperineal resection, one with low anterior resection, and one with fulguration alone. There were no recurrences in the 16 patients who had excision of the polypectomy site. If excluded, recurrence was 11 percent (4/37). Patients were examined at three month intervals for the first two years and every six months thereafter. There have been no cancer-related deaths. CONCLUSIONS: Transanal endoscopic microsurgical resection of pT1 rectal cancers yields low recurrence rates. Close follow-up permits curative salvage for those that do recur. Transanal excision remains a viable option. PMID- 16362802 TI - Effects of exopolysaccharide-producing probiotic strains on experimental colitis in rats. AB - PURPOSE: Inflammatory bowel disease is suggested to result from a dysregulated immune response toward intestinal microflora, which may be restored by probiotic therapy based on the concept of healthy microflora. Ideal probiotic bacteria may be beneficial in inflammatory bowel disease; however, the mechanism of action and the clinical efficacy of probiotic usage are still unclear. In the present study, the effect of exopolysaccharide producing probiotics was evaluated on an experimental colitis model in rats. METHODS: Colitis was induced by intracolonic administration of acetic acid. Then, rats were treated daily with two probiotic strains, Lactobacillus delbrueckii subsp. bulgaricus B3 strain (exopolysaccharide of 211 mg/l: high-EPS group) or Lactobacillus delbrueckii subsp. bulgaricus A13 strain (EPS of 27 mg/l: low-EPS group), which were given into the stomach. The non-colitis-fed control group was only treated with high-exopolysaccharide strain. The model-control and control groups were treated only with tap water. Rats were killed after a seven-day treatment period. Disease activity was quantified by use of histologic scores and colonic myeloperoxidase activity, which is a marker of neutrophil infiltration during inflammation. RESULTS: The enhanced inflammatory response was accompanied by a higher level of myeloperoxidase activity in the colitis group. Histologic scores of colonic damage and myeloperoxidase activity were lower in both probiotic-treated groups compared with those of the colitis control group (P<0.001), although the mentioned scores improved significantly more in the high-EPS group than in the low-EPS group (P<0.001). CONCLUSIONS: Exopolysaccharide-producing probiotics significantly attenuate experimental colitis, which may be mediated by exopolysaccharide in a dose-dependent manner. Therefore, exopolysaccharide producing probiotics may be a promising therapeutic role in inflammatory bowel disease. PMID- 16362803 TI - Randomized, controlled trial of anal electrical stimulation for fecal incontinence. AB - PURPOSE: Anal electric stimulation has been described as effective for fecal incontinence in several case series, but no study has addressed possible mechanism of benefit. We wished to examine whether anal electric stimulation, using an anal probe electrode, used on a daily basis at home for eight weeks, in the absence of any adjunctive exercises or advice, would improve symptoms of fecal incontinence and anal sphincter pressures when compared with "sham" electric stimulation. METHODS: Ninety patients (9 males, 81 females), with median age of 55 (range, 30-77) years were randomized, 47 to active anal stimulation at 35 Hz and 43 to "sham" stimulation at 1 Hz. Outcome measures included a one-week bowel diary, symptom questionnaire, manometry, and patients' evaluation of outcome. RESULTS: Seventy patients completed the study. On an intention-to-treat analysis, there was no difference between the two groups on any of the outcome measures after eight weeks. Of those who completed stimulation, 44 (63 percent) felt the stimulation had improved their continence. Those with intact anal sphincters were not likely to rate their change more positively than those with sphincter disruption (P=0.71). Median patient rating of bowel control increased from 3 of 10 before stimulation to 5 of 10 after stimulation (P=0.001). CONCLUSIONS: Eight weeks of anal electric stimulation was rated by patients as having improved their bowel control to a modest extent. There was no statistically significant difference detected between the groups, suggesting that 1 Hz was as effective as 35 Hz. This raises the possibility that the main effect is not sphincter contraction but sensitization of the patient to the anal area, or simply the effect of intervening per se. Home electric stimulation is a relatively cheap and generally well-tolerated therapy in the conservative treatment of fecal incontinence. PMID- 16362804 TI - Defecation disorders: a French population survey. AB - PURPOSE: Despite frequent occurrence, functional defecation disorders and related conditions have been infrequently reported in population studies. This study was designed to assess symptoms, lifestyle-behavioral changes, and medical care seeking related to functional defecation disorders in a large household community survey. METHODS: A large household community survey was conducted in 10,000 individuals aged 15 years or older. A mailed questionnaire was used to assess ten common anorectal complaints; frequency, association, impact on quality of life, and medical care seeking were quantified. RESULTS: Evaluation was obtained in 7,196 patients (3,455 males). During the previous 12-month period, 2,097 patients (29.1 percent) experienced functional defecation disorders: outlet constipation and fecal incontinence were reported in 22.4 and 16.8 percent respectively. Compared with patients with no anorectal complaint, patients with functional defecation disorders had a different gender status (females, 63.3 vs. 47.6 percent; P<0.01). Based on symptom severity, functional defecation disorders were perceived as the main anorectal complaint in 1,192 patients. In this group, emptying difficulties, unsatisfied defecation, gas, and fecal incontinence occurred at least once per month in 71.6, 56.1, 77.9, and 49 percent respectively: 66.6 percent with outlet constipation and 85.6 percent with incontinence revealed impairment in quality of life. Incontinent patients more frequently avoided medical care than those complaining of outlet constipation (67.4 vs. 46.4 percent; P<0.01). CONCLUSIONS: Functional defecation disorders concerns at least one of four French individuals. Outlet constipation and fecal incontinence frequently occur in association. Despite a low rate of patients seeking care, symptoms often are severe and related to quality of life impairment. PMID- 16362805 TI - Chemoprevention of colorectal cancer. AB - Colorectal cancer is a major public health concern in all developed countries. Despite decades of advances in the treatment and prevention of colorectal cancer, it remains the second most common cause of cancer death. However, the optimal method for early detection remains unknown and patient compliance with screening recommendations remains poor. This has led to the development of complementary strategies, such as chemoprevention to reduce morbidity and mortality from colorectal cancer. Chemoprevention is defined as the use of specific pharmacologic or nutrient agents to prevent, reverse, or inhibit the process of carcinogenesis. This review was designed to discuss the most promising agents in colorectal chemoprevention. PMID- 16362810 TI - Post-epidemic serosurvey of West Nile fever in Israel. PMID- 16362811 TI - Crohn disease obstruction of the biliopancreatic limb in a patient operated for biliopancreatic diversion for morbid obesity. AB - Although rare, patients with biliopancreatic diversion for morbid obesity will be subject to obstruction of the biliopancreatic limb. This condition is commonly due to postoperative adhesions and intussusception and usually presents with misleading clinical and radiographic features that can delay the diagnosis and the treatment. We recently encountered a patient with obstruction of the biliopancreatic limb due to involvement from Crohn disease. We report this case to highlight the clinical and imaging findings of this rare condition. PMID- 16362809 TI - Characterization of muscle-regulatory gene, MyoD, from flounder (Paralichthys olivaceus) and analysis of its expression patterns during embryogenesis. AB - Specification and differentiation of skeletal muscle cells are driven by the activity of genes encoding members of the myogenic regulatory factors (MRFs). In vertebrates, the MRF family includes MyoD, Myf5, myogenin, and MRF4. The MRFs are capable of converting a variety of nonmuscle cells into myoblasts and myotubes. To better understand their roles in fish muscle development, we isolated the MyoD gene from flounder (Paralichthys olivaceus) and analyzed its structure and patterns of expression. Sequence analysis showed that flounder MyoD shared a structure similar to that of vertebrate MRFs with three exons and two introns, and its protein contained a highly conserved basic helix-loop-helix domain (bHLH). Comparison of sequences revealed that flounder MyoD was highly conserved with other fish MyoD genes. Sequence alignment and phylogenetic analysis indicated that flounder MyoD, seabream (Sparus aurata) MyoD1, takifugu (Takifugu rubripes) MyoD, and tilapia (Oreochromis aureus) MyoD were more likely to be homologous genes. Flounder MyoD expression was first detected as two rows of presomitic cells in the segmental plate. From somitogenesis, MyoD transcripts were present in the adaxial cells that give rise to slow muscles and the lateral somitic cells that give rise to fast muscles. After 30 somites formed, MyoD expression decreased in the somites except the caudal somites, coincident with somite maturation. In the hatching stage, MyoD was expressed in other muscle cells and caudal somites. It was detected only in muscle in the growing fish. PMID- 16362812 TI - Rapid CT diagnosis of acute appendicitis with IV contrast material. AB - The purpose of this study was to determine the sensitivity and specificity of computed tomography (CT) without administration of oral contrast in confirming suspected acute appendicitis. One hundred seventy-three patient studies were retrieved by a computer-generated search for the word "appendicitis" in radiology reports. Patients presenting to the emergency department over an 8-month period were examined for acute abdominal pain or suspected acute appendicitis. IV contrast-enhanced CT scans of the abdomen and pelvis were obtained without oral or rectal contrast. Criteria for diagnosis of acute appendicitis included a dilated appendix (>6 mm), periappendiceal inflammation, or abscess. Final diagnoses were established with surgical/clinical follow-up, histopathological analysis or both. The standard time (1 h) for the administration of oral contrast prior to the CT scan was eliminated. Fifty-nine CT diagnoses were made of acute appendicitis, 56 of which were histologically verified and three of which resulted in another diagnosis. One hundred fourteen CT diagnoses were negative for appendicitis. This corresponds to a sensitivity of 100% and specificity of 97%, a positive predictive value of 95%, and a negative predictive value of 100%. CT with IV contrast is sensitive and specific for the confirmation or exclusion of acute appendicitis. By eliminating the time required to administer oral contrast, the diagnosis might be made more rapidly. PMID- 16362813 TI - Adult plant and seedling resistance to powdery mildew in a Triticum aestivum x Triticum militinae hybrid line. AB - In the progeny of a cross between the common wheat cultivar Tahti and Triticum militinae, a member of the timopheevii group of tetraploid wheats, several hybrid lines were selected that are characterized by improved seedling and adult plant resistance (APR) to powdery mildew. An F2 single-seed descendant mapping population segregating for seedling resistance and APR to powdery mildew was analysed for the identification of quantitative trait loci (QTL). The main QTL responsible for APR was detected on the long arm of chromosome 4A tightly linked to the Xgwm160 locus on a T. militinae translocation explaining up to 54% of phenotypic variance. The same translocation influenced seedling resistance to powdery mildew upon inoculation of plants with a synthetic population of Blumeria graminis DC. f. sp. tritici, and explained 28-33% of the phenotypic variance. PMID- 16362814 TI - Self-monitoring of blood glucose in type 2 diabetes and long-term outcome: an epidemiological cohort study. AB - AIMS/HYPOTHESIS: The aim of this study was to obtain epidemiological data on self monitoring of blood glucose (SMBG) in type 2 diabetes and to investigate the relationship of SMBG with disease-related morbidity and mortality. METHODS: The German multicentre Retrolective Study 'Self-monitoring of Blood Glucose and Outcome in Patients with Type 2 Diabetes' (ROSSO) followed 3,268 patients from diagnosis of type 2 diabetes between 1995 and 1999 until the end of 2003. Endpoints were diabetes-related morbidity (non-fatal myocardial infarction, stroke, foot amputation, blindness or haemodialysis) and all-cause mortality. SMBG was defined as self-measurement of blood glucose for at least 1 year. RESULTS: During a mean follow-up period of 6.5 years, 1,479 patients (45.3%) began SMBG prior to an endpoint and an additional 64 patients started SMBG after a non-fatal endpoint. Interestingly, many patients used SMBG while being treated with diet or oral hypoglycaemic drugs (808 of 2,515, 32%). At baseline, the SMBG cohort had higher mean fasting blood glucose levels than the non-SMBG cohort (p<0.001), suggesting that insufficient metabolic control was one reason for initiating SMBG. This was associated with a higher rate of microvascular endpoints. However, the total rate of non-fatal events, micro- and macrovascular, was lower in the SMBG group than in the non-SMBG group (7.2 vs 10.4%, p=0.002). A similar difference was found for the rate of fatal events (2.7 vs 4.6%, p=0.004). Cox regression analysis identified SMBG as an independent predictor of morbidity and mortality, with adjusted hazard ratios of 0.68 (95% CI 0.51-0.91, p=0.009) and 0.49 (95% CI 0.31-0.78, p=0.003), respectively. A better outcome for both endpoints was also observed in the SMBG cohort when only those patients who were not receiving insulin were analysed. CONCLUSIONS/INTERPRETATION: SMBG was associated with decreased diabetes-related morbidity and all-cause mortality in type 2 diabetes, and this association remained in a subgroup of patients who were not receiving insulin therapy. SMBG may be associated with a healthier lifestyle and/or better disease management. PMID- 16362815 TI - Peptide YY levels are decreased by fasting and elevated following caloric intake but are not regulated by leptin. AB - AIMS/HYPOTHESIS: Peptide YY (PYY) is a gut-derived hormone that has been shown to reduce short-term food intake in animals and humans. It has been proposed that deficiency of PYY contributes to obesity in humans. However, the physiology of PYY regulation by factors such as caloric restriction, or by other molecules important in energy homeostasis, e.g. leptin, remains to be fully elucidated. MATERIALS AND METHODS: We evaluated the effect on PYY levels of: (1) caloric ingestion (a mixed meal) in five healthy normal-weight subjects; (2) fasting for 2 or 3 days in eight lean men and seven lean women respectively; and (3) recombinant human leptin administration at physiological replacement and pharmacological doses. RESULTS: PYY levels increased 50% after a mixed meal (p=0.01), and short-term complete fasting for 2 or 3 days decreased leptin and PYY levels to 20-30% and 40-60% of baseline, respectively (both p<0.05). However, recombinant human leptin administration at physiological doses to restore the fasting-induced decrease of leptin levels and at pharmacological doses over the short term had no effect on PYY levels. CONCLUSIONS/INTERPRETATION: PYY increases after meal ingestion and decreases after fasting in a manner consistent with a meal-related signal of energy homeostasis. Importantly, circulating levels of this gut-secreted molecule are independent of regulation by leptin over the short term. These findings contribute towards our understanding of the homeostatic systems that regulate appetite in humans, including the possible redundancy of gastrointestinally secreted and adipocyte-secreted signals. This may be of importance for the future development of medications to treat obesity. PMID- 16362816 TI - Suitability of a fully 13C isotope labeled internal standard for the determination of the mycotoxin deoxynivalenol by LC-MS/MS without clean up. AB - Very often, the accuracy of quantitative analytical methods for the determination of mycotoxins by liquid chromatography (LC)-mass spectrometry (MS) and LC-MS/MS is limited by matrix effects during the ionization process in the MS source. Stable isotope labeled standards are best suited to correct for matrix effects and to improve both the trueness and the precision of analytical methods employing LC-MS and LC-MS/MS. This paper describes the successful use of fully 13C isotope labeled deoxynivalenol [(13C15)DON] as an internal standard (IS) for the accurate determination of DON in maize and wheat by LC electrospray ionization MS/MS. To show the full potential of (13C15)DON as IS, maize and wheat extracts were analyzed without further cleanup. Subsequent to calibration for the LC-MS end determination, DON was quantified in matrix reference materials (wheat and maize). Without consideration of the IS, apparent recoveries of DON were 29+/ 6% (n=7) for wheat and 37+/-5% (n=7) for maize. However, the determination of DON in the reference materials yielded 95+/-3% (wheat) and 99+/-3% (maize) when (13C15)DON was used as an IS for data evaluation. PMID- 16362817 TI - LST1 and NCR3 expression in autoimmune inflammation and in response to IFN-gamma, LPS and microbial infection. AB - Many genes in the central region of the major histocompatibility complex (MHC) encode proteins involved in immune and inflammatory responses. In this study, we have further characterized two genes in the MHC class IV region, leucocyte specific transcript (LST) 1 and natural cytotoxicity-triggering receptor 3 (NCR3) (also known as 1C7 and natural killer (NK)p30). The specific function of LST1 is not known, although expression analysis and functional data suggest an immunomodulatory role. The LST1 gene undergoes extensive alternative splicing, giving rise to both membrane-bound (encoded by exon 3) and soluble isoforms. The NCR3 protein is involved in NK-mediated cytotoxicity and plays a role in NK/dendritic cell crosstalk. Expression of these genes was examined, by real-time reverse transcriptase-polymerase chain reaction, in autoimmune-induced inflammation, specifically rheumatoid-arthritis-affected blood and synovium, and in response to stimulation with inflammatory mediators and bacterial agents. The expression of LST1, specifically splice variants encoding soluble isoforms and NCR3, was increased in rheumatoid-arthritis-affected blood and synovium and was associated with more severe inflammation in the synovium. Furthermore, both genes were significantly up-regulated in response to lipopolysaccharide, interferon (IFN)-gamma and bacterial infection. These findings suggest that NCR3 and soluble isoforms of LST1 may play a role in inflammatory and infectious diseases. PMID- 16362818 TI - Exploiting the genetic diversity of Beauveria bassiana for improving the biological control of the coffee berry borer through the use of strain mixtures. AB - Beauveria bassiana is an entomopathogen widely used to control the coffee berry borer in Colombia, as part of an Integrated Pest Management strategy. Traditionally, the development of fungal insect pathogens as biocontrol agents in crop pests has been oriented towards the selection and formulation of elite clonal strains. Instead, we explored the potential application of genetic diversity in B. bassiana by determining the effect of strain mixtures on coffee berry borer mortality compared to clonal isolates. Genomic DNA from 11 strains was characterized using internal transcribed spacers and beta-tubulin sequences as well as amplified fragment length polymorphism markers. Cluster analysis produced three genetic groups and confirmed the low but significant intraspecific genetic diversity present among the strains. Single strain virulence towards the coffee berry borer under laboratory conditions, using 1x10(6) conidia ml(-1), ranged between 89.9 and 57.5%. All the inoculations with mixtures resulted in coinfection events. Combinations of genetically similar strains showed no significant differences when their virulences were compared. However, mixtures of genetically different strains led to both antagonism and synergism. The lowest virulence percentage (57%) was obtained by putting together the most virulent strain of each group, contrary to the highest virulence percentage (93%) that resulted from mixing the three least virulent strains. The results indicate the promising potential of designing strain mixtures as an alternative for the biocontrol of Hypothenemus hampei and other pests and provide tools for the understanding of the ecological dynamics of entomopathogen populations under natural conditions. PMID- 16362819 TI - Optimisation of a 2-D gel electrophoresis protocol for the human-pathogenic fungus Aspergillus fumigatus. AB - Aspergillus fumigatus is the most important airborne fungal pathogen causing life threatening infections in immunosuppressed patients. One of the important questions concerning A. fumigatus is the identification of pathogenicity determinants. To obtain a comprehensive overview about the proteins produced at different physiological conditions that are related to the infectious process a proteomic approach has been applied. Here, 2-D gel electrophoresis for filamentous fungi was optimised concerning removal of interfering compounds, protein extraction and separation methods. A trichloroacetic acid-based precipitation method of proteins with their subsequent solubilisation by the use of a combination of CHAPS with a second sulfobetaine detergent gave the best results. The optimised protocol was evaluated by the analysis of the proteomes of A. fumigatus grown on two different carbon sources, i.e., glucose and ethanol. Carbon catabolite repression has not been studied in detail at the protein level in A. fumigatus yet. In addition, growth on ethanol leads to activation of the glyoxylate cycle which was shown to be essential for pathogenesis in bacteria and fungi. In A. fumigatus, differential patterns of enzymes of the gluconeogenesis, glyoxylate cycle and ethanol degradation pathway during growth on glucose and ethanol were observed. PMID- 16362821 TI - A finite element model investigation of gunshot injury. AB - Multiple gunshot suicide can be a controversial subject mainly because of wrong opinions concerning immediate incapacitation or alleged backwards hurling. For the last 20 years, experts in medicine and physics have tried to demonstrate what really happens during a gunshot wound. Different methods have been used to achieve this aim such as basic physics or the use of empirical evidence. In this paper, using a finite element model of the human head, we demonstrate that no incapacitation or backwards hurling can occur from a gunshot fired between the eyes which did not enter the cerebrum. PMID- 16362820 TI - Organization of interphase chromatin. AB - The organization of interphase chromatin spans many topics, ranging in scale from the molecular level to the whole nucleus, and its study requires a concomitant range of experimental approaches. In this review, we examine these approaches, the results they have generated, and the interfaces between them. The greatest challenge appears to be the integration of information on whole nuclei obtained by light microscopy with data on nucleosome-nucleosome interactions and chromatin higher-order structures, obtained in vitro using biophysical characterization, atomic force microscopy, and electron microscopy. We consider strategies that may assist in the integration process, and we review emerging technologies that promise to reduce the "resolution gap." PMID- 16362822 TI - Factors to keep in mind when introducing virtual microscopy. AB - Digitization of glass slides and delivery of so-called virtual slides (VS) emulating a real microscope over the Internet have become reality due to recent improvements in technology. We have implemented a virtual microscope for instruction of medical students and for continuing medical education. Up to 30,000 images per slide are captured using a microscope with an automated stage. The images are post-processed and then served by a plain hypertext transfer protocol (http)-server. A virtual slide client (vMic) based on Macromedia's Flash MX, a highly accepted technology available on every modern Web browser, has been developed. All necessary virtual slide parameters are stored in an XML file together with the image. Evaluation of the courses by questionnaire indicated that most students and many but not all pathologists regard virtual slides as an adequate replacement for traditional slides. All our virtual slides are publicly accessible over the World Wide Web (WWW) at http://vmic.unibas.ch . Recently, several commercially available virtual slide acquisition systems (VSAS) have been developed that use various technologies to acquire and distribute virtual slides. These systems differ in speed, image quality, compatibility, viewer functionalities and price. This paper gives an overview of the factors to keep in mind when introducing virtual microscopy. PMID- 16362823 TI - Pathologic examination of the axillary sentinel lymph nodes in patients with early-stage breast carcinoma: current and resolving controversies on the basis of the European Institute of Oncology experience. AB - Several controversial aspects of sentinel lymph node biopsy (SLNB) for patients with early-stage, node-negative breast carcinoma have been dealt with and resolved in the past decade since its introduction. Unfortunately, however, there is still no consensus on how best to examine sentinel lymph nodes (SLN) histologically. As a consequence, the protocols for SLN examination are remarkably variable in different institutions, leading to a very poor reproducibility of the data stemming from investigations on series of patients whose SLNs have been evaluated according to diverse protocols. Patient outcomes, however, can be optimised only by standardization of the whole procedure of SLNB, with particular reference to the histopathologic scrutiny. Lack of a standardized histopathologic protocol likely derives also from the uncertainties about the clinical implications of minimal lymph node involvement (isolated tumour cells and micrometastases) with regard both to the risk of additional metastases to non sentinel lymph nodes of the same basin and to the prognostic value for patients' survival. This review aims at highlighting some of the controversial issues of the histopathologic examination of the SLNs, including the number of sections and cutting intervals, the use of immunohistochemistry and the role of molecular biology assays. PMID- 16362824 TI - Novel and recurrent rearrangements in the CFTR gene: clinical and laboratory implications for cystic fibrosis screening. AB - Because standard techniques used to detect mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene do not detect single or multiple exonic rearrangements, the importance of such rearrangements may be underestimated. Using an in-house developed, single-tube, semi-quantitative fluorescent PCR (SQF PCR) assay, we analyzed 36 DNA samples submitted for extensive CFTR sequencing and identified ten samples with rearrangements. Of 36 patients with classic CF, 10 (28%) harbored various deletions in the CFTR gene, accounting for 14% of CF chromosomes. A deletion encompassing the CFTR promoter and exons 1 and 2 was detected in a sample from one proband, and in the maternal DNA as well. In another family, a deletion of the promoter and exon 1 was detected in three siblings. In both of these cases, the families were African American and the 3120+1G > A splice site mutation was also identified. These promoter deletions have not been previously described. In a third case, a deletion of exons 17a, 17b, and 18 was identified in a Caucasian female and the same mutation was detected in the paternal DNA. In the other seven cases, we identified the following deletions: exons 2 and 3 (n = 2); exons 4, 5, and 6a; exons 17a and 17b; exons 22 and 23; and exons 22, 23, and 24 (n = 2). In our series, the frequency of CFTR rearrangements in classic CF patients, when only one mutation was identified by extensive DNA sequencing, was >60% (10/16). Screening for exon deletions and duplications in the CFTR gene would be beneficial in classic CF cases, especially when only one mutation is identified by standard methodologies. PMID- 16362826 TI - [Abstracts. der Kongress der Deutschen Gesellschaft fur Psychiatrie, Psychotherapie und Nervenheilkunde. 23-26 November 2005. Berlin, Deutschland]. PMID- 16362825 TI - Peptidoglycan recognition proteins Pglyrp3 and Pglyrp4 are encoded from the epidermal differentiation complex and are candidate genes for the Psors4 locus on chromosome 1q21. AB - Psoriasis is a common inflammatory skin disease caused by genetic and environmental factors, including bacterial and viral infections. Since the skin is in constant contact with commensal and pathogenic microorganisms, we examined well-supported psoriasis genetic linkage intervals to identify genes encoding innate immune pattern recognition proteins that may play a role in pathogenesis. Two peptidoglycan recognition proteins, Pglyrp3 and Pglyrp4, are localized to the Psors4 locus on chromosome 1q21 in a gene cluster known as the epidermal differentiation complex (EDC). We show that these genes are expressed in the skin as well as in germinal centers in the tonsil. We tested 13 SNPs in or near these genes for association with psoriasis in two independent patient collections: a family-based patient set comprised of 375 individuals from 101 families, and a case-control patient collection of 282 patients with moderate to severe psoriasis and 192 healthy controls. In the family-based analysis, several SNPs in the Pglyrp3-Pglyrp4 locus show association with psoriasis (0.01 < P < 0.05). Multiple SNP haplotypes incorporating Pglyrp3 and Pglyrp4 SNPs also show significant association in the transmission disequilibrium test (TDT; P < 0.01). In the case control test, none of the SNPs that we tested show association with psoriasis when analyzed in single-SNP or haplotype-based tests. The discordance between the TDT and case-control results suggests that the two populations are significantly different in disease etiology, that the polymorphism responsible for the Psors4 linkage is elsewhere in the Pglyrp locus, or that the causative Psors4 polymorphism is in a location near but not in the Pglyrp locus. These data are consistent with previous reports of association of psoriasis with genes on 1q21, and suggest a role for Pglyrps in skin biology. PMID- 16362827 TI - Coordinated expression and immunogenicity of an outer membrane protein from Salmonella enterica serovar Typhi under iron limitation, oxidative stress and anaerobic conditions. AB - Successful pathogens overcome the environmental stresses by the coordinated expression of various genes and eventually proteins. Since, the surface of the microbe is likely to come in contact with the host initially, an attempt was made to identify the outer membrane proteins (OMPs), if any, which may get expressed under more than one environmental conditions simulating the in vivo ones. In the present study, Salmonella enterica serovar Typhi was grown under iron-limited, oxidative stress as well as anaerobic conditions and the OMP profiles were compared. A 69 kDa OMP was found to express with enhanced intensity under the selected stress conditions in comparison to normal conditions. The phenotypic similarity among the proteins was assessed on the basis of their molecular weight, cross reactivity and HPLC. The protein expressed under oxidative stress and anaerobic conditions reacted with the antibodies raised against iron regulated outer membrane protein (IROMP), indicating the sharing of at least some of the epitopes. A single peak observed after subjecting the pooled 69 kDa protein sample and appearance of a single band on SDS-PAGE thereafter, confirmed the purity and phenotypic similarity of the 69 kDa OMP. Reactivity of pooled 69 kDa protein with 85% of sera from typhoid patients revealed the in vivo expression of this protein. The results of this study indicate the coordination of this phenotype under iron stress, oxidative stress and anaerobic conditions. In view of the expression of the 69 kDa protein under the selected stress conditions and their in vivo immunogenicity, these findings may be relevant for the better understanding of the host-microbe interactions and for the further development of diagnostic and preventive strategies. PMID- 16362829 TI - Adrenal steroids in adrenomyeloneuropathy. Dehydroepiandrosterone sulfate, androstenedione and 17alpha-hydroxyprogesterone. AB - Adrenoleukodystrophy (ALD) and its adult variant adrenomyeloneuropathy (AMN) are X-linked diseases associated with a deficiency in the peroxisomal degradation of saturated very long chain fatty acids (VLCFA) resulting in an accumulation of VLCFA in the central and peripheral myelin, the adrenal cortex and the testis. Adrenal insufficiency with clinical hypocortisolism occurs in approximately two thirds of the patients with AMN. We studied the circulating adrenal hormones 17alpha-hydroxyprogesterone (17alpha-OHP), androstenedione and dehydroepiandrosterone sulphate (DHEAS) in 63 male AMN patients (age 17-65 years) and the DHEAS serum levels in 95 healthy male controls (age 30-65 years). 34 of the patients presented with the phenotype of only spinal cord and peripheral nerve disability without hypocortisolism, 29 of the patients presented with the phenotype of either additional hypocortisolism or Addison's syndrome only. Normal 17alpha-OHP concentrations were found in all patients with no significant difference between patients without and with hypocortisolism (6.07 +/- 0.61 nmol/l and 4.76 +/- 0.37 nmol/l). Androstenedione concentration was significantly (p < 0.01) lower in patients with hypocortisolism (2.99 +/- 0.65 pmol/l versus 5.71 +/- 0.68 pmol/l). As serum levels of DHEAS are agedependent we divided the two groups into two subgroups each (subgroup one: age 17-40 years, subgroup two: age 41-65 years). The DHEAS concentration of patients without and with hypocortisolism was significantly (p < 0.01) lower in both subgroups (1. 4.35 +/- 0.84 micromol/l, n = 15, 2. 15 +/- 0.28 micromol/l, n = 19; 1. 1.90 +/- 0.57 micromol/, n = 21, 2. 0.96 +/- 0.29 micromol/l, n = 8) compared to controls (1. 9.0 +/- 0.96 micromol/l; 2. 5.21 +/- 0.25 micromol/l). In conclusion, androstenedione and DHEAS serum concentrations are subnormal in all AMN patients and may therefore serve as sensitive markers of the adrenal function in adrenomyeloneuropathy. PMID- 16362831 TI - Pallido-pyramidal syndrome with blepharospasm and good response to levodopa. PMID- 16362828 TI - Diagnostic investigation and multidisciplinary management in motor neuron disease. AB - Amyotrophic lateral sclerosis (ALS) is the most common adult-onset motor neuron disease. ALS is a progressive neurodegenerative disorder, involving motor neurons in the cerebral cortex, brainstem and spinal cord, presenting with a combination of upper and lower motor neuron signs. Etiology remains undetermined, although a multifactorial origin is widely accepted including genetic factors, auto immunity, oxidative stress, glutamate excitotoxicity and abnormal neurofilament aggregation. The absence of specific diagnostic testing, and variable clinical presentations make the diagnosis of ALS challenging, relying upon correlation of clinical, electrophysiological and neuroimaging data. The disease is relentlessly progressive, with dysarthria, dysphagia, tetraparesis, and respiratory insufficiency due to ongoing respiratory muscle paresis. There is no specific treatment for ALS. Riluzole, a glutamate antagonist, is the only FDA approved drug for ALS, but has only a modest effect on survival. The multiplicity and progressiveness of the disabilities in ALS, highlights the need for a coordinated multidisciplinary rehabilitation program managing symptoms, respiratory care, dysphagia and nutrition, dysarthria and communication, physical and occupational therapy. The main goals are to prolong independence, prevent complications and improve quality of life. PMID- 16362830 TI - Determinants of autonomic dysfunction in idiopathic Parkinson's disease. AB - OBJECTIVES: To determine demographic or disease-related factors that may influence the severity of autonomic dysfunction in idiopathic Parkinson's disease (IPD). METHODS: 532 patients with IPD aged between 55 and 75 years were included. Severity of autonomic dysfunction was assessed using a 9-item autonomic dysfunction score (ADS). In addition, several demographic factors (e. g. age, gender, comorbidities) and disease- related (e. g. motor stage, disease duration, antiparkinsonian therapy) factors were recorded. A group of 67 age-matched healthy volunteers served as a control group. Demographic and clinical data of this cross-sectional survey were analyzed by a logistic stepwise regression model to determine independent predictors of autonomic dysfunction. RESULTS: IPD patients showed significantly higher ADS values than controls, even in the youngest age groups and in mild disease stages. Hoehn & Yahr (H&Y) stage, disease duration, age at onset and various therapy combinations all showed significant correlations with ADS. However, stepwise logistic regression revealed that age (OR 10.71; CI 7.17-16.0) and arterial hypertension (OR 3.05; CI 1.66-5.58) were the only independent risk factors associated with autonomic dysfunction. Linear regression indicated that ADS increases with age in controls as well as in patients, but with a significantly steeper slope in the latter. CONCLUSIONS: Autonomic dysfunction as an inherent feature of IPD is present already in early disease stages. According to a logistic regression model, the severity of autonomic dysfunction in IPD is primarily related to demographic but not to disease-related factors. This and the differences in predictors for motor versus autonomic decline may indicate at least partly independent neurodegenerative processes. PMID- 16362832 TI - Familial autosomal dominant cortico-basal degeneration with the P301S mutation in the tau gene: an example of phenotype variability. PMID- 16362833 TI - Vascular parkinsonism. Comments on Sibon et al. J Neurol (2004) 251:513-524. PMID- 16362839 TI - Spinocerebellar ataxia type 4 (SCA4): Initial pathoanatomical study reveals widespread cerebellar and brainstem degeneration. AB - Spinocerebellar ataxia type 4 (SCA4), also known as 'hereditary ataxia with sensory neuropathy', represents a very rare, progressive and untreatable form of an autosomal dominant inherited cerebellar ataxia (ADCA). Due to a lack of autopsy cases, no neuropathological or clinicopathological studies had yet been performed in SCA4. In the present study, the first available cerebellar and brainstem tissue of a clinically diagnosed and genetically-confirmed German SCA4 patient was pathoanatomically studied using serial thick sections. During this systematic postmortem investigation, along with an obvious demyelinization of cerebellar and brainstem fiber tracts we observed widespread cerebellar and brainstem neurodegeneration with marked neuronal loss in the substantia nigra and ventral tegmental area, central raphe and pontine nuclei, all auditory brainstem nuclei, in the abducens, principal trigeminal, spinal trigeminal, facial, superior vestibular, medial vestibular, interstitial vestibular, dorsal motor vagal, hypoglossal, and prepositus hypoglossal nuclei, as well as in the nucleus raphe interpositus, all dorsal column nuclei, and in the principal and medial subnuclei of the inferior olive. Severe neuronal loss was seen in the Purkinje cell layer of the cerebellum, in the cerebellar fastigial nucleus, in the red, trochlear, lateral vestibular, and lateral reticular nuclei, the reticulotegmental nucleus of the pons, and the nucleus of Roller. In addition, immunocytochemical analysis using the anti-polyglutamine antibody 1C2 failed to detect any polyglutamine-related immunoreactivity in the central nervous regions of this SCA4 patient studied. In view of the known functional role of affected nuclei and related fiber tracts, the present findings not only offer explanations for the well-known disease symptoms of SCA4 patients (i.e. ataxic symptoms, dysarthria and somatosensory deficits), but for the first time help to explain why diplopia, gaze-evoked nystagmus, auditory impairments and pathologically altered brainstem auditory evoked potentials, saccadic smooth pursuits, impaired somatosensory functions in the face, and dysphagia may occur during the course of SCA4. Finally, the results of our immunocytochemical studies support the concept that SCA4 is not a member of the CAG-repeat or polyglutamine diseases. PMID- 16362841 TI - [Psychopathology and personality patterns in the first-degree relatives of bipolar patients]. AB - OBJECTIVE: This is a cross-sectional study designed to assess psychopathology and personality patterns in a group of high-risk subjects for bipolar disorder compared to a control group. As high-risk subjects first-degree relatives of bipolar patients were selected. METHOD: Ninety-five first-degree relatives of 54 bipolar patients and 93 first-degree relatives of 54 subjects without any psychiatric disorder were recruited in the study. Control subjects were matched to bipolar patients according to age, gender and educational status. SADS-L (Schedule for Affective Disorders and Schizophrenia-Lifetime Version) was used both to ascertain the psychiatric status of the patient and control subjects, and to evaluate the psychopathology in probands' and controls' relatives. MMPI-2 (Minnesota Multiphasic Personality Inventory-2) profiles of relatives of patient and control groups were compared as well. RESULTS: In the relatives of bipolar patients the SADS-L diagnoses of hypomania, minor depression and schizotypal personality were statistically more prevalent than in the relatives of the control group. MMPI-2 profiles of both relatives of bipolar patients and controls were within "normal" range, whereas relatives of patients were more defensive in disclosing psychopathology. Any specific profile characteristic for relatives of bipolar patients could not be described. CONCLUSION: Minor mental disorders were more prevalent in the relatives of bipolar patients group. A personality pattern specific to high risk group for bipolar disorder couldn't be detected. PMID- 16362842 TI - [A study on alcohol use among working youth who are continuing apprenticeship training]. AB - OBJECTIVE: Considering both burden of entering working life and engaging in adult roles in terms of being employed, working youth forms a specific group to be handled regarding alcohol use problems. In this study it was aimed to investigate prevalence and patterns of alcohol use among youth in apprenticeship schools in central Ankara. METHOD: A survey was done to investigate presence of alcohol use problems among students of five apprenticeship schools in Ankara using CAGE Questionnaire and a questionnaire form including more detailed questions about alcohol use. RESULTS: Prevalence of life-time use of alcohol was 37.3% and prevalence of alcohol use more than once was 24.3%. Ten per cent of students with a history of alcohol use told that they went on drinking during last year. Proportion of students who continued drinking during the last year and getting at least 2+ score from CAGE was found as 4.5%. CONCLUSION: Prevalence of clinically significant alcohol use is quite high among working youth. Thus, in order to plan preventive measures, risk factors for alcohol use problems should be considered in detail in working youth. PMID- 16362843 TI - [Combination therapy using sertraline with sleep deprivation and light therapy compared to sertraline monotherapy for major depressive disorder]. AB - OBJECTIVE: Bright light therapy is effective and well tolerated in seasonal affective disorder and some studies reported an antidepressant effect of bright light also in non-seasonal depression. On the other hand, total sleep deprivation leads to a rapid and marked improvement of mood in 60% of depressed patients. Combinations of antidepressant medication with those somatic therapies are generally indicated. The aim of this study was to compare the efficacy of the combination of sertraline and partial sleep deprivation or light therapy with sertraline monotherapy in the treatment of major depression. METHOD: Thirty-seven patients with major depressive disorder were randomly allocated into 3 treatment groups. Thirteen were treated with sertraline and late partial sleep deprivation, 13 with sertraline and bright light therapy and 11 sertraline monotherapy as a control group. Outcome measures included daily (first 15 days) and weekly Hamilton Rating Scale for Depression and biweekly Hamilton Anxiety Rating Scale. RESULTS: Partial sleep deprivation group improved significantly and more rapidly. Accelerated treatment response was shown in sleep deprivation group that improvement was observed after the third day. Bright light and sleep deprivation combinations with sertraline were more effective than sertraline monotherapy for accompanied anxiety in depression. CONCLUSION: Late partial sleep deprivation in combination with sertraline can accelerate and increase the treatment response in non-seasonal major depressive disorder. PMID- 16362844 TI - [Validity and reliability of the Turkish version of the Wender Utah Rating Scale for attention-deficit/hyperactivity disorder in adults]. AB - OBJECTIVE: To determine reliability and validity of Turkish form of the 25 item Wender Utah Rating Scale (WURS), which is designed as an aid for diagnosis of Attention-Deficit/Hyperactivity Disorder (ADHD) in adults. METHOD: The scale was administered to 59 patients who were diagnosed as having ADHD according to DSM-IV criteria after comprehensive psychiatric and neuropsychological assessments in outpatient clinics of University of Ankara Medical School, Department of Psychiatry. Control groups consisted of 59 patients with depression, 44 patients with bipolar affective disorder in remission, and 145 healthy controls. Subjects who were illiterate and younger than 18 years of age were excluded from the study. WURS was readministered to 63 of the healthy controls after 4 weeks. RESULTS: Principal components analysis revealed 5 factors explaining 61.3% of the variance. The factors were labeled as Irritability, Depression, School Problems, Behavioral Problems/Impulsivity and Attentional Deficits. Mean factor scores of ADHD group was higher than all groups except Depression factor. Turkish form of WURS demonstrated excellent internal consistency (Cronbach's alpha= 0.93), and the test-retest coefficient for the WURS (total score) was 0.81. Item-total score correlations varied between 0.31-0.75. A cut off score of 36 or higher identified 82.5% of the adults with ADHD (sensitivity), 90.8% of the healthy controls (specificity), 66% of patients with depression, and 64.3% of patients with bipolar affective disorder correctly. CONCLUSION: Turkish form of the WURS is a reliable and valid scale in assessing childhood symptoms in adults for ADHD. However overlapping mood disorder items lowers specificity of the scale. PMID- 16362845 TI - [Development and reliability study of the adolescent inhalant use severity scale]. AB - OBJECTIVE: To develop a scale in order to investigate the severity of inhalant use in adolescents and to conduct reliability and validity studies of this scale. METHOD: A questionnaire was developed by conducting interviews with two groups: professionals working with adolescent inhalant users and inhalant-using adolescents. This questionnaire was developed into a 21-item scale. This was applied by two different interviewers to 50 adolescents at Bakirkoy Mental Health Hospital Inhalant Use Research and Treatment Center for Children and Adolescents. Two other professionals applied the Visual Analog Scale to the same 50 adolescents to evaluate the severity of inhalant use. The correlation between the two interviewer scores and the scores obtained from the Visual Analog Scale and the 21-item scale was evaluated. Addicts and non-addicts were evaluated using SCID-I and the scores of addicts and non-addicts were compared. Test re-test reliability of the scale was evaluated by applying the scale in the 10-15 days following the first administration. RESULTS: The correlation between the questionnaire scores and the Visual Analog Scale scores was adequate (0.72). The mean scores of addicts were higher than those of non-addicts (p<0.001). The Cronbach alpha value was 0.84. Three items had no internal consistency and the same items' interrater and test re-test reliability correlations were low. CONCLUSION: It is thought that when the 3 inconsistent items are removed the 18 item Yeniden Inhalant Use Severity Scale (YUKUD) is a reliable and valid measure to evaluate the severity of inhalant use in adolescents and that it can be used in related research. PMID- 16362846 TI - [The neurobiology of hallucinations]. AB - OBJECTIVE: Although hallucinations are among the cardinal features of schizophrenia, the mechanisms underlying these symptoms have not been fully understood yet. In this review our objective is to summarize the studies about the neurobiology of auditory hallucinations in schizophrenia and discuss their significance for understanding schizophrenia. METHOD: This is a retrospective literature review and the relevant studies within the last 10 years are included. For this purpose Pubmed search engine and 'hallucination, schizophrenia, neurobiology' key words are used. RESULTS: Hallucinations cause sensory modality specific activation in cerebral areas involved in normal sensation. A disturbance in perception of speech seems to have a central role in occurrence of auditory hallucinations. Anatomically, auditory hallucinations appear to involve primary and association cortices, Broca's and Wernicke's areas, subcortical, paralimbic, limbic regions, ventral striatum and thalamus. Furthermore they are suggested to be associated with the dysmodulation of the information flow from ventral striatum to thalamus and cortex caused by increased dopaminergic activity in mesolimbic pathway. CONCLUSION: The validity of the models which are proposed to explain neurobiology of hallucinations in schizophrenia should be tested by new studies. The difficulties regarding the assessment and measurement of subjective mental phenomena are important impacts in studying and understanding the neural correlates of hallucinations. Functional neuroimaging, cognitive and molecular biological studies will hopefully enhance our knowledge about this disease. PMID- 16362847 TI - [The relationship between mood disorders and temperament, character and personality]. AB - The terms temperament, character and personality have been used almost synonymously despite their different meanings. Hippocratic physicians conceptualized illness, including melancholia, in dimensional terms as an out growth of premorbid characteristics. In modern times, full-scale application of this dimensional concept to psychiatric disorders led Kraepelin, Schneider and Kretschmer to hypothesize that the 'endogenous psychoses are nothing other than marked accentuation of normal types of temperament'. Akiskal's 'soft-bipolarity' and 'affective temperaments' concepts and Cloninger's psychobiological model of temperament and character, which includes four temperament and three character dimensions, are examples of this dimensional approach from the last two decades. Hypotheses concerning the relationship between personality disorders and mood disorders have been described, but it is likely that a single unitary model would not adequately capture the complexity inherent in the relationship between mood and personality disorders. The DSM multiaxial approach to diagnosis encourages the clinician to distinguish state (Axis I) from trait (Axis II) features of mental disorders. Categorical systems like DSM have been criticised because of their inability to mention temperament, character and personality features. In this review, examples of dimensional approaches to mood disorders are given and discussed under the influence of temperament, character and personality disorders. For this purpose, literature from 1980 to 2004 has been reviewed through Pub/med, using the following key words. PMID- 16362848 TI - [Frontoparietal hypoperfusion in Capgras syndrome: a case report and review]. AB - Capgras syndrome (CS) is characterized by the delusional belief that a person, usually very close to the patient, has been replaced by a double who is physically very similar to the original. CS is relatively rare, occurring predominantly in course of schizophrenia, particularly of the paranoid sub-type, and less frequently in association with schizoaffective and affective disorder. Recent years have witnessed a sharp increase in the number of published CS cases with an organic etiology; however, CS was considered to have its origins in psychodynamic conflict. We present a patient with the CS and brain SPECT findings whom without psychiatric disorder. As an evaluation of brain SPECT, there have been found a significantly decreased blood flow in bilateral parietal regions and slightly decreased blood flow in bilateral posterior frontal regions. Cerebral dysfunction is proposed to be a central role in CS development. Unilateral right hemisphere lesions occur more frequently than the left; however, the majority of CS cases show bilateral involvement. Pathology involves many parts of the brain, most notably frontal and parietal cortex. Our findings support that frontoparietal dysfunction could be important in the pathogenesis of CS. We reviewed the neurobiology of CS and discussed our findings in this article. CS studies will give a better understanding of the neurobiological basis of psychotic experiences and may contribute to develop a paradigm on researches about other psychotic disorders. PMID- 16362849 TI - Verification of the left ventricular ejection fraction from gated myocardial perfusion studies (GSPECT). AB - INTRODUCTION: The perfusion study (which may be obtained using SPECT or GSPECT technology within six hours of administration of the radionuclide Tc-99m-MIBI) reflects the regional blood supply to the left ventricular [LV] myocardium at the time of radionuclide administration (i.e. at rest, at peak exercise, or at peak vasodilatation), while the values of EF, EDV, and ESV measured using GSPECT are parameters of LV contractility at the time of image acquisition (i.e. at rest or in a nearresting state following exercise or vasodilatation). Planar radionuclide ventriculography [RNV] is, however, considered to be the most accurate method for calculating LVEF. AIMS: The main goal of the study was to compare the values of EF obtained by the most frequently used method, GSPECTQGS, and the reference method, RNV - taking into consideration various clinical scenarios (presence or absence of LV dilatation) and various conditions under which GSPECT was recorded (at rest, post-exercise, or post-dipyridamole). METHODS: Two hundred patients (145 males) aged 58+/-11 (18-80) with previously confirmed (n=166, of whom 108 had a history of myocardial infarction) or suspected (n=34) coronary artery disease were included in the study. Ranges of normal values for EF, EDV, and ESV were established based on a group of 26 'normal' subjects. LV dilatation was defined as an EDV >127 ml (at rest, measured by QGS) - this was present in 88 patients. Myocardial perfusion studies were obtained using GSPECT following administration of Tc-99m-MIBI at rest (all patients), as well as one hour after treadmill exercise (138 patients) or dipyridamole administration (48 patients). The resting RNV was conducted within three weeks of the GSPECT exam. The EF values obtained by QGS and RNV were compared for patients with and without LV dilatation. EF, EDV, and ESV values obtained by QGS were compared for resting patients, post-exercise, and post-dipyridamole. RESULTS: 1. The GSPECT EF values calculated using QGS software, at rest or one hour after treadmill exercise or dipyridamole administration, demonstrated, for the study population as a whole, a significant, strong correlation with the results obtained by the reference method, RNV (correlation coefficient, r> or =0.86). The correlation was stronger in patients with LV dilatation than in those without. Both in patients with and without LV dilatation the correlation of EF with RNV was slightly weaker for postexercise (relative to resting) and post-dipyridamole (relative to post exercise) measurements. 2. QGS tended to underestimate the absolute values of EF, as compared to RNV. 3. In post-exercise and post-dipyridamole measurements, relative to the resting measurements (in patients with previously diagnosed or suspected coronary artery disease) the mean values of EF were lower while EDV and ESV were higher. CONCLUSIONS: In order to complement data on myocardial perfusion, the GSPECT-QGS technique should be optimally used to calculate LV contractility parameters at rest (as opposed to post-exercise or post dipyridamole), and include a range of normal values for EF, EDV, and ESV, obtained using QGS. Of note, EF measurements by GSPECT are more accurate for dilated than non-dilated ventricles. PMID- 16362851 TI - Cytokines and heart rate variability in patients with chronic heart failure. AB - INTRODUCTION: Heart rate variability (HRV) analysis is a non-invasive method of assessment of the autonomic nervous system's effects on heart function. In chronic heart failure (CHF), decreased HRV correlates with the progression of the disease. It is also known that in CHF increased levels of proinflammatory cytokines are present. Because these molecules are believed to influence the nervous system at both the central and peripheral levels, their potential role in HRV reduction in the course of CHF has been proposed. AIM: The study was designed to verify potential relations between cytokines and HRV parameters in CHF patients. The concept of the study was driven by the recognition of controversies in this field and the paucity of published reports. METHODS: Forty-four patients with CHF and stable NYHA class I-IV symptoms and 15 healthy controls were enrolled in the study. Time-domain HRV analysis was performed based on of 24-hour Holter ECG monitoring. Plasma concentrations of soluble TNFalpha receptors sTNF RI and sTNF-RII and interleukin 6 (IL-6) were measured using commercially available ELISA kits (Quantikine, RD Systems). RESULTS: In patients with CHF, HRV indices included in the analysis were significantly decreased, and the levels of cytokines increased in comparison with the control group. In the whole study population, both in the CHF patients and the control group, significant negative correlations were observed between sTNF-RI level and long-term HRV indices such as SDNN (r=-0.44; p=0.0006), SDANN (r=-0.44; p=0.0005) and short-time index SDNNI (r=-0.37; p=0.004). Similar negative correlations were found between sTNF-RII level and SDNN (r=-0.35; p=0.007), SDANN (r=-0.34; p=0.01), and SDNNI (r=-0.31; p=0.02), as well as between IL-6 level and SDNN (r=-0.41; p=0.001), SDANN (r= 0.44; p=0.0005) and SDNNI (r=-0.34; p=0.009). CONCLUSIONS: Significant negative correlations between TNF-alpha soluble receptors sTNF-RI, sTNF-RII and IL-6 levels and time-domain HRV parameters were observed in the study. Because the results of investigations conducted so far do not elucidate the cause-effect relationship, further studies are needed to clarify the mechanisms of HRV depression in CHF and the role of cytokines in this severe clinical condition. PMID- 16362853 TI - Invasive treatment of coronary artery disease in octogenarians. AB - INTRODUCTION: There are many patients aged over 80 years among those hospitalised for coronary artery disease (CAD). The unanswered question is whether invasive treatment of such patients is effective and safe. AIM: To assess and compare one year clinical outcomes after percutaneous coronary angioplasty (PTCA) and surgical coronary artery bypass grafting (CABG) in patients aged over 80 years and in younger patients. METHOD: There were 63 patients aged over 80 years suffering from CAD who underwent either PTCA or CABG. The control group consisted of 40 patients aged 60-65 years treated in the same way. Data on medical history, cardiovascular risk factors, and angiographic findings were analysed. The potential risks of the procedures, post-procedural complications as well as the clinical status at the end of one-year follow-up were evaluated. RESULTS: There were 24 surgical revascularisation procedures and 39 PCIs performed in the very old patients. Stable angina was found in 29 cases, unstable angina in 19 and acute myocardial infarction in 15 patients. There were three in-hospital deaths and 18 periprocedural complications were noted. During the one-year follow-up period six deaths occurred, persistent or recurrent angina was found in 11 patients after PCI and two after CABG. There were no deaths in the control group and the incidence of minor complications was similar to the senile group. In younger patients who underwent CABG, CCS class at one year was lower than in the very old ones. CONCLUSIONS: The invasive treatment of coronary artery disease in octogenarians is feasible with satisfactory results and acceptable procedural risks. PMID- 16362855 TI - Fractional flow reserve assessment to determine the indications for myocardial revascularisation in patients with borderline stenosis of the left main coronary artery. AB - INTRODUCTION: Reliable assessment of clinical significance of borderline angiographic lesions found within the left main coronary artery (LM) is often impossible. Measurement of fractional flow reserve (FFR) is commonly used to verify borderline stenoses of the coronary arteries. However, the usefulness of FFR measurements has been validated only for arteries other than the LM. AIM: Evaluation of the measured FFR value in determination of the indications for myocardial revascularisation in borderline LM stenosis. METHODS: The study involved 38 patients aged 55+/-9 years (range 41-74 years) with isolated borderline LM stenosis. Each patient had the measurement of FFR performed during intravenous adenosine infusion at a dose of 140 microg/kg/min. Patients were referred for revascularisation if FFR was <0.75. RESULTS: The mean LM stenosis in quantitative coronary angiography (QCA) was 45+/-10%. FFR<0.75 was found in 18 (47%) patients, whereas 20 (53%) subjects had FFR < or =0.75. In subjects with FFR <0.75 QCA showed significantly lower minimal lumen diameters (MLD) at the site of stenosis (1.84+/-0.45 vs 2.24+/-0.49, p=0.014). Additionally, a significant correlation was found between FFR and MLD (r=0.59, p<0.001). The mean clinical follow-up was 2 years (range 1-3 years). There were two (11%) fatal events in patients with FFR < or =0.75 who underwent CABG. One (5%) patient with FFR >0.75 underwent elective CABG due to progression of LMN stenosis. Moreover, one (5%) patient experienced myocardial infarction not related to borderline stenosis of the LM. CONCLUSIONS: The measurement of FFR confirms the clinical significance of stenosis only in half of the patients with borderline isolated lesion of the left main coronary artery. Withdrawal from intervention in patients with FFR > or =0.75 is safe and is associated with favourable clinical outcomes in two-year follow-up. PMID- 16362857 TI - [Diagnosis and treatment of chronic heart disease. Guidelines of the European Society of Cardiology. Revision 2005]. PMID- 16362859 TI - [Sleep apnea syndrome as a cause of secondary hypertension. A case report]. AB - A case of a 51-year old man, suffering from drug-resistant hypertension, complaining of hypersomnia and fatigue during the day, is presented. In the course of diagnostic procedures the diagnosis of sleep apnea syndrome was established. Continuous positive airway pressure (CPAP) therapy was successfully started. Examination carried out 3 months later revealed good response to pharmacological treatment with normal levels of blood pressure. PMID- 16362861 TI - [Myocardial infarction in the peripartum period in a 42-year-old multipara with normal coronary arteries]. AB - Acute myocardial infarction (AMI) during pregnancy and puerperium is very rare. With a growing number of older women giving birth, AMI may become more common complication of pregnancy. We present a 42-year-old multipara with coagulation disorders in pregnancy who suffered from AMI in the peripartum period, documented by ECG, cardiac markers and echocardiography, with normal coronary arteries on angiogram. In conclusion, AMI in pregnancy may be different entity than AMI complicating coronary artery disease. It seems that the main causes of AMI in this population may not be atherosclerotic plaques, but reversible factors, i.e. vessel spasm or coagulation disorders. This may explain why in our patient symptoms of AMI were dramatic in spite of a small area of necrosis and normal coronary angiograms. PMID- 16362862 TI - [Stentgraft implantation for the treatment of coronary artery aneurysm in a patient with acute coronary syndrome]. PMID- 16362864 TI - [RF ablation in a patient with recurrent post-infarction ventricular tachycardia requiring multiple cardioverter-defibrillator interventions]. AB - A case of a 51 year old patient with a history of myocardial infarction (MI) and recurrent ventricular tachycardia (VT) is presented. Three months after MI the patient underwent coronary angioplasty and one year later received prophylactic implantable cardioverter-defibrillator (ICD) due to complex ventricular arrhythmias, detected on Holter ECG monitoring, and depressed left ventricular ejection fraction. Later on the patient started to experience palpitations and ICD shocks during physical activity (cycling). Interrogation of the ICD memory showed appropriate shocks due to slow (160 betas/min) VT. The device was reprogrammed and new antitachycardia pacing (ATP) algorithms were enabled, however, it occurred proarrhythmic due to the ATP-induced acceleration of VT rate. Finally, in April 2005 he received 37 appropriate ICD shocks during a few hours. The patient was selected for RF ablation and underwent successful procedure with the use of the electro-anatomical CARTO mapping system. PMID- 16362865 TI - [Severe right-ventricular heart failure due to malignant pericardial mesothelioma]. AB - A case of a 63-year old female with symptomatic advanced right-ventricular (RV) heart failure due to malignant pericardial mesothelioma is presented. Echocardiography revealed that RV failure was due to the tumour-induced compression of the right atrium and not due to metastatic mesothelioma involving pericardial sac. PMID- 16362866 TI - [Electrocardiogram of a patient with a history of ventricular fibrillation - peculiar right bundle branch block or atypical Brugada syndrome?]. PMID- 16362868 TI - [Sirolimus-eluting vs paclitaxel-eluting stents -- the trials confirm differences in clinical outcome]. PMID- 16362870 TI - [Emergency medicine -- set off and change of an interdisciplinary discipline]. PMID- 16362871 TI - [Requirements and expectations for optimal volume replacement]. AB - A volume replacement should compensate a reduction in the intravascular volume and counteract a hypovolemia so that hemodynamics and vital functions can be maintained. For this therapy, a physiologically-based solution comprising both osmotically and colloid osmotically active components should be administered. A consensus is proposed for this purpose which takes into consideration the following aspects: The optimum colloid, the questionable use of albumin, the physiological electrolyte pattern encompassing sodium, potassium, chloride and phosphate and their contributions to osmolality, an eventual addition of glucose, the physiological acid-base status with bicarbonate or alternately with metabolisable anions, and the importance of a clear declaration of all ingredients. The consensus distinguishes between compulsory requirements derived from evidence-based medicine and physiological data and the potential expectations of an optimal volume replacement, including well-grounded wishes and aspirations for the future. PMID- 16362872 TI - [Emergency room management of multiple injured patients. A multislice computed tomography orientated treatment algorithm]. AB - INTRODUCTION: In the treatment of the seriously injured patient, time is crucial. Clarke et al. showed that in the hypotensive patient with blunt abdominal trauma and bleeding a delay in the surgical treatment over 90 min increases mortality of 1 % every 3 minutes. So called trauma algorithms are used as systematic approach to the seriously injured patient that can be easily reviewed and practiced. One impact of this algorithm is to shorten time in the emergency room and may improve outcome of the patient. The patient outcome is directly related to the length of time between the injury and the beginning to proper definitive care. OBJECTIVE: Are improvements still necessary and possible in the care of the seriously injured patient? New technical developments like multislice-CT are able to shorten time until definitive care. The practicability of an multislice-CT orientated treatment algorithm was evaluated. METHODS: After integration of an multislice-CT into the emergency room the treatment of the seriously injured patient (ISS >16) was changed. Therefore an algorithm which puts the CT diagnostics in the first minutes of the clinical care was developed. RESULTS: 50 patients were treated following the new algorithm. The time in the emergency room decreased from 87 min to 36 min. CONCLUSION: This algorithm requires a differing from previous surgical standards, but it proved to be timesaving, effective and practicable. It makes the beginning of the definitive care possible within the first 40 min after patient arrival in the emergency room. PMID- 16362873 TI - [How to assess the severity of the multi-system trauma in the emergency-room -- a critical review]. AB - For more than 30 years various scoring systems have been used as a method to asses the injury severity of multi-system trauma. They have been used as a tool for triage, for emergency-room quality management, for educational reasons or, nowadays, in order to evaluate the cost effectiveness of either a complete hospital or a single department. This review provides the knowledge to score multi-system traumatized patients after their first treatment in the emergency room. It shows advantages, disadvantages, and limitations of various traumascores which includes knowledge about the used parameters, the type of validation, the medical system in their country of origin, and of course their mathematical background. Relevant examples like physiological trauma scores (GCS, RTS, MEES), anatomic, injury pattern based scores (ISS, NISS, ICISS), biological aspects, mixed scoring systems (PTS, TRISS, ASCOT) and future developments ("Rixen pattern", RISC) are illustrated. These scores are explained in detail and discussed as to their practicability. PMID- 16362874 TI - [Characteristics of repeated emergency physician use]. AB - OBJECTIVE: Repeated use of emergency physicians might result in excessive costs and has discouraging effects on emergency department staff. To our knowledge, no data are available on the characteristic of repeated use of emergency physicians (EP) in the German emergency medical services. Accordingly, this retrospective survey focused on emergency cases on-the-scene, which necessitated EP intervention. METHODS: Following data from the local EMS computerised database (Medlinq, EASY, Hamburg, Germany) were retrospectively analysed for the period from January to December of 2002: total number of EP interventions, person who engaged the emergency call, number of repeat EP-users with two to three calls, number of frequent EP-users with more than three calls, their sex, age, NACA. Probably inappropriate repeat users were defined as patients with no apparent underlying distress on-the-scene, lack of preclinical findings which could explain the symptoms initially mentioned, and refuse of hospital referral by the patient. The rate of repeated inappropriate EP use was defined as the ratio of the number of repeated inappropriate use over the total number of EP-engagements in the year 2002. RESULTS: 6064 EP interventions were analysed. The rate of the repeated use was 15.5 % (939/6064) including 3.6 % (218/6064) frequent users. The main medical causes (80 %) of repeated EP use were cardiovascular, neurological, respiratory, and psychiatric. The rate of repeated inappropriate EP use was 0.2 % (11/6064). CONCLUSION: The rate of repeated inappropriate EP use in this middle sized town was low. More than four fifths of all repeated calls were for cardiovascular, respiratory, neurological, and psychiatric distress. PMID- 16362875 TI - [Advance directives in the prehospital setting -- emergency physicians' attitudes]. AB - OBJECTIVE: The German physician based emergency medical system (EMS) might confront physicians with advance directives in the field. A multi-question survey was used to evaluate emergency physicians' experience with advance directives in the prehospital setting and to assess their attitudes towards forms and statements of advance directives. METHODS: A questionnaire was mailed to the members of the Association of Emergency Physicians of Northern Germany ("AGNN"), an interest group of emergency physicians, in 2001. RESULTS: 511 emergency physicians (50,4 % of the AGNN members) filled in the questionnaire completely and sent it back for evaluation. 75 % of the participants were working as emergency physicians at present, 72 % had emergency experiences of more than 5 years. One third had previously dealt with advance directives in the prehospital setting. 77 % of these physicians thought advance directives generally helpful. Nevertheless 88 % based their management on the context of the individual circumstances (e. g. emergency conditions, underlying diseases, expected prognosis), only 7 % said they would always exactly follow the statements of the directive. In the view of the emergency physicians the advance directive should contain information on cardiopulmonary resuscitation (CPR: 88 %), intensive care treatment (75 %) and preclinical emergency treatment (55 %). Information on underlying diseases (87 %) and a legal substitute (84 %) should be contained as well. As formal requirements, 47 % of the physicians wanted the family doctor to be involved, 49 % desired a notary authenticity confirmation, additionally or solely. Pragmatically, the advance directive should be kept with the personal documents (84 %). A regular reconfirmation was deemed necessary (twice to once a year: 64 %). The current legal situation was regarded as unclear by 81 % of the emergency physicians, 85 % favored a unique, officially authorized type of directive. CONCLUSION: The high number of returned questionnaires shows the importance of the topic "advance directives" for emergency physicians. Despite some practical and legal problems, a big majority of the experienced emergency physicians in this survey thought the advance directives in the prehospital setting to be helpful. A clear statement on resuscitation as well as simplification of the many existing types of directives are the most essential requirements demanded by the emergency physicians. A solution could be the creation of an extra "emergency advance directive". PMID- 16362876 TI - [Non-invasive determination of cardiac output by continuous wave Doppler in air rescue service]. AB - BACKGROUND: Determination of cardiac output (CO) enables to assess the hemodynamic situation as well as to administer optimal catecholamine therapy especially in critically compromised patients with hemodynamic instability. Invasive determination of CO is possible via a Swan-Ganz-catheter with its associated risk of implantation in the hospital. Using the Doppler technique, we evaluated the feasibility of the USCOM-system for non-invasive CO determination in preclinical emergency medicine in air rescue service. METHODS: In 32 patients (17 months to 92-years-old) cardiac output was determined non-invasively (USCOM) at the scene and during the helicopter transport at Christoph 4, based at Hannover Medical School. Simultaneously, blood pressure, ECG and oxygen saturation were determined. Non-invasive CO was assessed by a suprasternal access aiming at the aorta ascendens. 19 patients were unconscious due to cardial and non-cardial reasons, and 13 were conscious (sepsis, status epilepticus, anaphylactic reaction). 7 patients were hemodynamically unstable. In three patients the monitor was used during interhospital transfer by helicopter. RESULTS: Non-invasively determined CO via the USCOM system was 4.8 +/- 0.7 l/min with a cardiac index of 2.4 +/- 0.3 l/m (2). Highest CO values were determined in a patient with sepsis and during a grand-mal-status in epilepsy (CO 8.2 l/min). All examinations were done by the same emergency physician of the emergency helicopter Christoph 4 immediately after arrival at the scene. The examination took on average 25 seconds. During the helicopter transport, several consecutive CO measurements were performed to assess volume and catecholamine therapy with increase of stroke volume after volume load with colloidal fluids. CONCLUSIONS: Using the USCOM system it is possible to determine the beat-to-beat cardiac output in air rescue service non-invasively. The emergency physician gains additional crucial hemodynamic information to diagnose and treat adequately by administration of volume load and catecholamines at the scene and during flight conditions. Further preclinical prospective trials are mandatory to elucidate the value of this novel device in emergency medicine. PMID- 16362877 TI - [Polytrauma with tension pneumothorax with inserted chest tube]. AB - The authors report a case of a 25-year-old woman with a polytrauma, caused by a free fall of 12 metres in suicidal intention. Following endotracheal intubation and mechanical ventilation by an emergency physician at the scene, the patient was delivered to the emergency room of an university hospital. An ultrasonic check of the abdomen revealed free fluid in the abdominal cavity, and a rupture of liver and spleen was suspected. Since breath sounds over the right lung were diminished, a chest tube was inserted immediately in the fifth intercostal space in the anterior axillary line. About 300 millilitres of blood were drained by the tube. Shortly thereafter, a laparotomy was performed, where spleen and liver rupture were confirmed and treated. After 60 minutes, the patient developed severe hypotension coupled with ventricular tachycardia and fibrillation, and resuscitation measures had to be initiated. Since breath sounds over the right lung were missing, a tension pneumothorax was suspected and a thoracotomy performed immediately. While huge amounts of air and blood were emerging from the thoracic cavity, a rupture of the right mainstem bronchus as well as of the right pulmonary artery and vena subclavia was identified. The chest tube was found dislocated into the subcutaneous tissue. Despite of open heart compression, application of adrenaline and noradrenaline and substitution of packed red blood cells and of crystalloid and colloid solutions, all resuscitation measures failed so that the patient died shortly after on the operation table. This case illustrates first the difficulties of an adequate thoracic trauma management, particularly, when clinical symptoms are discrete, second the problems of the insertion and control of a chest tube, and third risks associated with wrong position or secondary dislocation which may include - as in our case - "masking" of severe injury patterns and delay of life-saving measures such as an immediate thoracotomy. In order to improve prognosis of patients with poly-/thoracic trauma, establishment of spiral-CT in emergency centres, routine bronchoscopy and safe handling of chest tubes may be helpful. PMID- 16362878 TI - [Cardiac arrest caused by an ecstasy intoxication]. AB - We report about a 19 years old man, suffering from an cardiac arrest (ventricular fibrillation) caused by an ecstasy intoxication. A supraventricular tachycardia was recorded on day three after resuscitation. No pathological findings were demonstrated by coronary angiography. An slow- fast- av -nodal- reentry- tachycardia (AVNRT) was detected and successfully treated by electrical ablation of the slow pathway during electrophysiological mapping. No severe neurological deficits were found in discharge from hospital. PMID- 16362879 TI - Community-associated methicillin-resistant Staphylococcus aureus: the way to the wound is through the nose. PMID- 16362880 TI - Prevalence of Staphylococcus aureus nasal colonization in the United States, 2001 2002. AB - BACKGROUND: Staphylococcus aureus is a common cause of disease, particularly in colonized persons. Although methicillin-resistant S. aureus (MRSA) infection has become increasingly reported, population-based S. aureus and MRSA colonization estimates are lacking. METHODS: Nasal samples for S. aureus culture and sociodemographic data were obtained from 9622 persons > or = 1 year old as part of the National Health and Nutrition Examination Survey, 2001-2002. After screening for oxacillin susceptibility, MRSA and selected methicillin-susceptible S. aureus isolates were tested for antimicrobial susceptibility, pulsed-field gel electrophoresis clonal type, toxin genes (e.g., for Panton-Valentine leukocidin [PVL]), and staphylococcal cassette chromosome mec (SCCmec) type I-IV genes. RESULTS: For 2001-2002, national S. aureus and MRSA colonization prevalence estimates were 32.4% (95% confidence interval [CI], 30.7%-34.1%) and 0.8% (95% CI, 0.4%-1.4%), respectively, and population estimates were 89.4 million persons (95% CI, 84.8-94.1 million persons) and 2.3 million persons (95% CI, 1.2-3.8 million persons), respectively. S. aureus colonization prevalence was highest in participants 6-11 years old. MRSA colonization was associated with age > or = 60 years and being female but not with recent health-care exposure. In unweighted analyses, the SCCmec type IV gene was more frequent in isolates from participants of younger age and of non-Hispanic black race/ethnicity; the PVL gene was present in 9 (2.4%) of 372 of isolates tested. CONCLUSIONS: Many persons in the United States are colonized with S. aureus; prevalence rates differ demographically. MRSA colonization prevalence, although low nationally in 2001-2002, may vary with demographic and organism characteristics. PMID- 16362881 TI - RNAIII-inhibiting peptide significantly reduces bacterial load and enhances the effect of antibiotics in the treatment of central venous catheter-associated Staphylococcus aureus infections. AB - BACKGROUND: Medical devices used in clinical practice are often associated with biofilm-associated staphylococcal infections. METHODS: An in vitro antibiotic susceptibility assay of Staphylococcus aureus biofilms using 96-well polystyrene tissue-culture plates was performed to test the effects of RNAIII-inhibiting peptide (RIP), ciprofloxacin, imipenem, and vancomycin. Efficacy studies were performed using a rat model of central venous catheter (CVC)-associated infection. Twenty-four hours after implantation, the catheters were filled with RIP (1 mg/mL). Thirty minutes later, rats were challenged, via the CVC, with 1.0 x 10(6) cfu of S. aureus strain Smith diffuse. The antibiotic-lock technique was begun 24 h later. RESULTS: Minimum inhibitory concentrations of antibiotics in biofilms were at least 4-fold higher than those against the freely growing planktonic cells. When they were first treated with RIP, the cells in biofilms became as susceptible to antibiotics as did planktonic cells. These data were confirmed by the in vivo studies. In particular, when CVCs were treated with both RIP and antibiotics, the biofilm bacterial load was further reduced to 1 x 10(1) cfu/mL, and bacteremia was not detected, suggesting that there was 100% elimination of bacteremia and a 6 log10 reduction in biofilm bacterial load. CONCLUSION: RIP significantly reduces bacterial load and enhances the effect of antibiotics in the treatment of CVC-associated S. aureus infections. PMID- 16362882 TI - The acute-phase response and serum amyloid A inhibit the inflammatory response to Acinetobacter baumannii Pneumonia. AB - BACKGROUND: Acinetobacter baumannii is an emerging pathogen in nosocomial pneumonia. Trauma and postsurgical patients display a profound acute-phase protein response and are susceptible to pneumonia. METHODS: To study the way in which the acute-phase response induced by sterile tissue injury influences pulmonary host defense, mice were injected subcutaneously with turpentine or saline in both hind limbs either 2 or 5 days before intranasal inoculation with A. baumannii. RESULTS: Turpentine-injected mice demonstrated strong increases in levels of the acute-phase proteins serum amyloid A (SAA) and serum amyloid P. The inflammatory response to A. baumannii was significantly impaired in turpentine injected mice, as shown by decreased local cytokine and chemokine levels, reduced neutrophil influx and lung myeloperoxidase activity, less pulmonary inflammation on histological examination, and lower total protein levels in their bronchoalveolar lavage fluid, which was associated with reduced bacterial clearance of A. baumannii. The late acute-phase protein response still caused lower pulmonary cytokine levels and neutrophil recruitment. Furthermore, previous injection of SAA, a major acute-phase protein, also reduced inflammatory responses to A. baumannii pneumonia. CONCLUSIONS: These data suggest that the acute-phase response and SAA inhibit the local inflammatory response to A. baumannii pneumonia, which may facilitate bacterial outgrowth. PMID- 16362883 TI - Preexposure to live Brugia malayi microfilariae alters the innate response of human dendritic cells to Mycobacterium tuberculosis. AB - BACKGROUND: Mycobacterium tuberculosis and helminth coinfection is highly prevalent, and the presence of helminths may modulate the Th1 response necessary for M. tuberculosis control. METHODS: Elutriated human monocytes, differentiated into dendritic cells (DCs) and macrophages, were exposed in vitro to live microfilariae (mf). The influence that mf had on M. tuberculosis infectivity, expression of cell surface molecules, and production of cytokines was determined. RESULTS: Compared with mf-unexposed, M. tuberculosis-infected cells, mf-exposed, M. tuberculosis-infected DCs had decreased expression of CD14, CD54, and human leukocyte antigen-DR, and mf-exposed, M. tuberculosis-infected macrophages had decreased expression of CD40. DCs that were mf exposed and M. tuberculosis infected produced more interleukin (IL)-1 beta than did mf-unexposed, M. tuberculosis-infected DCs. Also, mf-exposed, M. tuberculosis-infected DCs and macrophages expressed less IL-10 and interferon (IFN)- alpha than did mf unexposed, M. tuberculosis-infected cells. When they were cultured with autologous CD4+ T cells, mf-exposed, M. tuberculosis-infected DCs were less capable of stimulating the production of IFN- gamma than were other DCs. Exposure of DCs to mf decreased the surface expression of DC-specific intercellular adhesion molecule-3 grabbing nonintegrin, a receptor required by M. tuberculosis for entry into DCs. CONCLUSIONS: Exposure to mf reduces a key receptor on the DC surface, which perhaps renders these cells less susceptible to infection with M. tuberculosis. Exposure to mf changes the surface expression of adhesion and costimulatory molecules on DCs and macrophages and alters their expression of cytokines and chemokines in a way that renders them less capable of immunologic responses. PMID- 16362884 TI - CD11b regulates recruitment of alveolar macrophages but not pulmonary dendritic cells after pneumococcal challenge. AB - Despite their close physical and functional relationships, alveolar macrophages (AMs) and pulmonary dendritic cells (pulDCs) have rarely been examined together in the context of infection. Using a nonlethal, resolving model of pneumonia caused by intranasal injection of Streptococcus pneumoniae, we demonstrate that AMs and pulDCs exhibit distinct characteristics during pulmonary inflammation. Recruitment of AMs and pulDCs occurred with different kinetics, and increased numbers of AMs resulted mainly from the appearance of a distinct subset of CD11b(High) AMs. Increased numbers of CD11b(High) and CD11b(Low) AMs, but not pulDCs, were recoverable from bronchoalveolar lavage fluid. CD11b expression on AMs was significantly increased by granulocyte-macrophage colony-stimulating factor but not by interleukin-10 or pathogen-associated stimuli. Finally, antibody blockade demonstrated that CD11b was critical for the recruitment of AMs, but not pulDCs, into the lung after pneumococcal challenge. These data demonstrate that there are significant differences between AM and pulDC responses to inflammatory pathogenic stimuli in vivo. PMID- 16362885 TI - Influenza-induced expression of indoleamine 2,3-dioxygenase enhances interleukin 10 production and bacterial outgrowth during secondary pneumococcal pneumonia. AB - BACKGROUND: Airway infection with influenza virus induces local expression of the tryptophan-catabolizing enzyme indoleamine 2,3-dioxygenase (IDO), which has been shown to enhance inflammatory mediator responses in vitro. Because secondary pneumococcal infections occurring shortly after recovery from influenza are associated with enhanced inflammatory responses, we hypothesized that IDO activity contributes to the enhanced response to bacterial challenges in mice previously infected with influenza virus. METHODS: On day 14 after influenza virus infection (with strain A/PR/8/34), C57Bl/6 mice were intranasally inoculated with 1 x 10(4) colony-forming units of S. pneumoniae (serotype 3). Matrix-driven delivery pellets that contained 70 mg of the IDO inhibitor 1-methyl DL-tryptophan (MeTrp) released over a period of 7 days were subcutaneously implanted 48 h before pneumococcal infection. RESULTS: MeTrp treatment resulted in a 20-fold reduction in pneumococcal outgrowth 48 h after bacterial inoculation. Remarkably, pulmonary levels of interleukin-10 and tumor necrosis factor-alpha were significantly reduced in mice treated with MeTrp. CONCLUSIONS: Our data suggest that IDO expression during influenza virus infection alters the inflammatory response and facilitates the outgrowth of pneumococci during secondary bacterial pneumonia. PMID- 16362886 TI - Colonic bacterial infection abrogates eosinophilic pulmonary disease. AB - Induction of immunity to one pathogen in the lungs modifies the microenvironment and alters immunopathological changes that result from a second, unrelated pulmonary infection. However, it is unclear whether immunity generated at distant sites also affects lung immune responses. Here, we show that infection with the gut-restricted bacterium Citrobacter rodentium modifies immunopathological changes that result from pulmonary Cryptococcus neoformans infection. Th2 cytokine-driven pulmonary eosinophilia induced by C. neoformans infection was reduced, and the enhanced Th1 cytokine environment afforded more-rapid clearance of the fungus in C. rodentium-immune mice. The activated and intraepithelial (CD103+) T cell populations that expand after C. neoformans infection were diminished in C. rodentium-immune mice. T cell cross-reactivity was absent, but cross-reactive antibodies were detected. It is of importance to the "hygiene hypothesis" that these data indicate that an immune response induced by a gut restricted pathogen can modify the immune outcome after pulmonary infection, suggesting that cell-phenotype modifications occur across mucosal sites. PMID- 16362887 TI - Pleiotrophic effects of 2 Enterococcus faecalis sagA-like genes, salA and salB, which encode proteins that are antigenic during human infection, on biofilm formation and binding to collagen type i and fibronectin. AB - BACKGROUND: We have shown previously that Enterococcus faecium SagA has broad spectrum binding to extracellular matrix (ECM) proteins. In the present study, 2 sagA-like genes, salA and salB, were identified in Enterococcus faecalis. METHODS: We compared the salA and salB mutants; their parental strain, OG1RF; and the salB-complemented strain for binding to ECM proteins and biofilm formation. RESULTS: The salB mutant (TX5123) grew more slowly but showed greater binding (approximately 10%-20% of cells bound) to fibronectin (FN) and collagen type I (CI) than did OG1RF (approximately 1% of cells bound) (P<.001). Although TX5123 showed decreased biofilm formation in tryptic soy broth plus 0.25% glucose (TSBG) (P<.001 vs. OG1RF), a marked increase in biofilm formation was shown by TX5123 but not by OG1RF when they were grown in TSBG plus horse serum (HS) or TSBG plus FN, and the increase was coincident with increased attachment and hydrophobicity of TX5123. Complementation of the salB mutant restored the wild-type phenotypes. CONCLUSIONS: Whether SalB expression is ever sufficiently low in vivo to enhance adherence to ECM proteins or the serum-elicited increase in biofilm formation seen with the salB mutant in vitro is not currently known, but it is a potential way in which this organism could increase its adherence and biofilm formation during infection. PMID- 16362888 TI - Group B Streptococcus crosses human epithelial cells by a paracellular route. AB - Colonization of the colon and vagina is thought to be important in the pathogenesis of group B Streptococcus (GBS) infection. However, little is known about the strategies used by GBS to translocate through the epithelial barrier during the onset of disease. We used differentiated epithelial cells grown on transwell inserts as a model of the epithelial barrier. Bacterial translocation occurred without a detectable decrease in transepithelial resistance. Whereas acapsular GBS was better able to adhere to and invade epithelial cells, the percentage of bacteria translocating across the epithelial monolayer was independent of the presence of the capsule. Transmission electron microscopy showed the intimate association of GBS with intercellular junctions and the capacity to cross the monolayer by a paracellular mechanism. This process consisted of an active and transient opening of cell junctions. Indeed, GBS was preferentially found along the cell perimeter, where it colocalized with junctional protein complexes. PMID- 16362889 TI - The duration of hypotension before the initiation of antibiotic treatment is a critical determinant of survival in a murine model of Escherichia coli septic shock: association with serum lactate and inflammatory cytokine levels. AB - BACKGROUND: This study was designed to examine the relationship between the timing of antibiotic treatment and both survival rates and hemodynamic/inflammatory correlates of survival in a murine model of Escherichia coli septic shock. METHODS: Surgical implantation of an E. coli (O18:K1:H7) laced, gelatin capsule-encased fibrinogen clot was used to generate a bacteremic model of murine septic shock. Survival duration, hemodynamic responses, and circulating serum tumor necrosis factor (TNF)-alpha , interleukin (IL)-6, and lactate levels were assessed in relation to increasing delays in or absence of antibiotic treatment. RESULTS: A critical inflection point with respect to survival occurred between 12 and 15 h after implantation. When initiated at or before 12 h, antibiotic treatment resulted in < or = 20% mortality, but, when initiated at or after 15 h, it resulted in >85% mortality. Physiologically relevant hypotension developed in untreated septic mice by 12 h after implantation. Values for heart rate differed between untreated septic mice and sham-infected control mice by 6 h after implantation, whereas values for cardiac output and stroke volume did not differ until at least 18-24 h after implantation. Antibiotic treatment initiated > or = 12 h after implantation was associated with persistence of increased circulating serum lactate, TNF- alpha , and IL-6 levels. CONCLUSIONS: The timing of antibiotic treatment relative to hypotension is closely associated with survival in this murine model of septic shock. Delay in antibiotic treatment results in the persistence of inflammatory/stress markers even after antibiotic treatment is initiated. PMID- 16362890 TI - Immunologic response to antiretroviral therapy in hepatitis C virus-coinfected adults in a population-based HIV/AIDS treatment program. AB - BACKGROUND: We sought to characterize the impact that hepatitis C virus (HCV) infection has on CD4 cells during the first 48 weeks of antiretroviral therapy (ART) in previously ART-naive human immunodeficiency virus (HIV)-infected patients. METHODS: The HIV/AIDS Drug Treatment Programme at the British Columbia Centre for Excellence in HIV/AIDS distributes all ART in this Canadian province. Eligible individuals were those whose first-ever ART included 2 nucleoside reverse transcriptase inhibitors and either a protease inhibitor or a nonnucleoside reverse transcriptase inhibitor and who had a documented positive result for HCV antibody testing. Outcomes were binary events (time to an increase of > or = 75 CD4 cells/mm3 or an increase of > or = 10% in the percentage of CD4 cells in the total T cell population [CD4 cell fraction]) and continuous repeated measures. Statistical analyses used parametric and nonparametric methods, including multivariate mixed-effects linear regression analysis and Cox proportional hazards analysis. RESULTS: Of 1186 eligible patients, 606 (51%) were positive and 580 (49%) were negative for HCV antibodies. HCV antibody-positive patients were slower to have an absolute (P<.001) and a fraction (P = .02) CD4 cell event. In adjusted Cox proportional hazards analysis (controlling for age, sex, baseline absolute CD4 cell count, baseline pVL, type of ART initiated, AIDS diagnosis at baseline, adherence to ART regimen, and number of CD4 cell measurements), HCV antibody-positive patients were less likely to have an absolute CD4 cell event (adjusted hazard ratio [AHR], 0.84 [95% confidence interval [CI], 0.72-0.98]) and somewhat less likely to have a CD4 cell fraction event (AHR, 0.89 [95% CI, 0.70-1.14]) than HCV antibody-negative patients. In multivariate mixed-effects linear regression analysis, HCV antibody-negative patients had increases of an average of 75 cells in the absolute CD4 cell count and 4.4% in the CD4 cell fraction, compared with 20 cells and 1.1% in HCV antibody-positive patients, during the first 48 weeks of ART, after adjustment for time-updated pVL, number of CD4 cell measurements, and other factors. CONCLUSION: HCV antibody-positive HIV-infected patients may have an altered immunologic response to ART. PMID- 16362891 TI - Lymphoproliferative response in primary human cytomegalovirus (HCMV) infection is delayed in HCMV transmitter mothers. AB - BACKGROUND: The T cell-mediated immune response to human cytomegalovirus (HCMV) after primary infection, as well as the determinants of intrauterine transmission, are poorly understood. METHODS: Sequential peripheral blood leukocyte samples from 74 pregnant women and 29 nonpregnant individuals with primary infection were examined for HCMV-specific CD4+ T cells by cytokine flow cytometry (CFC) and lymphoproliferative response (LPR) analysis. Immunological results for 19 transmitter and 21 nontransmitter mothers were compared. RESULTS: Comparison of CFC and LPR analysis results showed that (1) there was no difference between pregnant and nonpregnant individuals; (2) HCMV-specific CD4+ T cells were detected by CFC, in the absence of an LPR to HCMV, in the great majority or the totality (according to different intervals) of samples collected from both pregnant and nonpregnant individuals during follow-up; and (3) LPR to HCMV was significantly (P<.001) lowered or delayed in transmitter mothers, compared with that in nontransmitter mothers. CONCLUSIONS: Pregnancy does not influence the HCMV-specific immune response. A dissociation between CFC response and LPR is commonly observed in patients with primary infections, and ad hoc studies aimed at understanding the mechanism(s) of the reduced LPR in transmitter mothers are warranted. PMID- 16362892 TI - Human leukocyte antigen concordance and the transmission risk via breast-feeding of human T cell lymphotropic virus type I. AB - OBJECTIVE: We examined the association between mother-to-child human T cell lymphotropic virus type I (HTLV-I) transmission and human leukocyte antigen (HLA) class I types. METHODS: In 1989, children born to HTLV-I-infected mothers in Jamaica were enrolled and prospectively evaluated for HTLV-I infection. HLA class I types in mothers and children were determined by DNA-based polymerase chain reaction methods. Associations between HLA class I types and transmission of HTLV I were analyzed using proportional-hazards regression models adjusted for the duration of breast-feeding. Transmission risk in children still breast-feeding at 12 months was determined using actuarial methods. RESULTS: Of 162 children, 28 (17%) became infected. After Bonferroni's adjustment for multiple comparisons, the transmission risk was not influenced by any specific HLA class type or the A2 supertype. However, compared with children who shared 3 HLA class I types with their mothers (the minimum number possible), the transmission risk increased 1.8 fold with 4 shared types and 3.0-fold with 5 or 6 shared types (Ptrend = .039; 1.75-fold increase for each additional concordant HLA type). This association was independent of maternal HTLV-I proviral level, antibody titer, and household income. CONCLUSIONS: We found a significant dose-response relationship between HTLV-I transmission via breast-feeding and mother-child HLA class I type concordance. Immunological interactions between a child's cells and maternal cells may influence the risk of HTLV-I infection by breast-feeding, perhaps because antigens on maternal cells are seen by the child as being "self." PMID- 16362893 TI - Lack of association between infection with a novel human coronavirus (HCoV), HCoV NH, and Kawasaki disease in Taiwan. AB - We investigated whether infection with a novel human coronavirus (HCoV), called "New Haven coronavirus" (HCoV-NH)--which is similar to and likely represents the same species as another novel HCoV, HCoV-NL63--is associated with Kawasaki disease (KD) in Taiwan. Fifty-three patients with KD were enrolled in our study. Serum, peripheral-blood mononuclear cells, nasopharyngeal aspirates, throat swabs, and rectal swabs from these patients were assayed for HCoV-NL63 by real time reverse-transcriptase (RT) polymerase chain reaction (PCR), and the throat swabs, nasopharyngeal aspirates, and rectal swabs were also assayed for HCoV-NH by RT-PCR. All PCR results were negative for both HCoV-NL63 and HCoV-NH; therefore, we did not find any association between HCoV-NH infection and KD in Taiwan. PMID- 16362894 TI - Multiple Epstein-Barr virus strains in patients with infectious mononucleosis: comparison of ex vivo samples with in vitro isolates by use of heteroduplex tracking assays. AB - Recent work using a heteroduplex tracking assay (HTA) to identify resident viral sequences has suggested that patients with infectious mononucleosis (IM) who are undergoing primary Epstein-Barr virus (EBV) infection frequently harbor different EBV strains. Here, we examine samples from patients with IM by use of a new Epstein-Barr nuclear antigen 2 HTA alongside the established latent membrane protein 1 HTA. Coresident allelic sequences were detected in ex vivo blood and throat wash samples from 13 of 14 patients with IM; most patients carried 2 or more type 1 strains, 1 patient carried 2 type 2 strains, and 1 patient carried both virus types. In contrast, coresident strains were detected in only 2 of 14 patients by in vitro B cell transformation, despite screening >20 isolates/patient. We infer that coacquisition of multiple strains is common in patients with IM, although only 1 strain tends to be rescued in vitro; whether nonrescued strains are present in low abundance or are transformation defective remains to be determined. PMID- 16362895 TI - Genetic variants in nonclassical major histocompatibility complex class I human leukocyte antigen (HLA)-E and HLA-G molecules are associated with susceptibility to heterosexual acquisition of HIV-1. AB - Human leukocyte antigen (HLA)-E and HLA-G molecules act as powerful modulators of the innate immune response. The present study shows that the HLA-E(G) genetic variant (the HLA-E*0103 allele) alone is significantly (P = .001) associated with a 4.0-fold decreased risk of human immunodeficiency virus 1 (HIV-1) infection in Zimbabwean women. Furthermore, women carrying the combination of the protective HLA-E(G) homozygote and HLA-G*0105N heterozygote genotypes had a 12.5-fold decreased risk of HIV-1 infection (P = .03), compared with women carrying neither genotype. These associations remained significant after adjustment was made for other significant sociodemographic risk factors for HIV prevalence in this population. In conclusion, HLA-E and HLA-G polymorphisms can independently and synergistically influence susceptibility to heterosexual acquisition of HIV-1. PMID- 16362896 TI - Antibody responses to hepatitis A virus vaccine in HIV-infected children with evidence of immunologic reconstitution while receiving highly active antiretroviral therapy. AB - BACKGROUND: Human immunodeficiency virus (HIV)-infected patients have weak responses to vaccines and may require revised immunization regimens. We investigated the safety and immunogenicity of 2 doses of hepatitis A virus (HAV) vaccine followed by a booster dose in HIV-infected children receiving stable highly active antiretroviral therapy. METHODS: A total of 235 children with CD4+ T cell percentages > or = 20% received 2 vaccine doses 24 weeks apart, and 117 received a third vaccine dose after 104 weeks. Anti-HAV antibody titers were measured at baseline and at 32, 104, and 112 weeks after the first vaccination. Subjects with antibody titers > or = 20 mIU/mL were defined as being seropositive. High and low antibody responses were defined as titers > or = 250 and <250 mIU/mL, respectively. RESULTS: Of 151 subjects who were HAV seronegative at baseline, 97% seroconverted after 2 vaccine doses, and 47% had low antibody responses. At 104 weeks, 90% of subjects had antibody titers > or = 20 mIU/mL, and those with low antibody responses were more likely to lose protective antibody titers. A third vaccine dose generated significantly higher antibody titers than those observed after the second vaccine dose. Undetectable HIV RNA at baseline was associated with higher anti-HAV antibody titers after the second vaccine dose. Antibody titers after the second and third vaccine doses were weakly correlated with CD4+ T cell percentages at the time when each vaccine dose was administered. In the 45 subjects who were HAV seropositive at baseline, responses to 2 and 3 vaccine doses were higher than those in subjects who were HAV seronegative at baseline, but the responses showed similar correlations. There were no serious adverse events associated with the vaccine. CONCLUSIONS: HIV-infected children with CD4+ T cell percentages > or = 20% responded better to the HAV vaccine if they had undetectable HIV RNA. The standard 2-dose immunization regimen generated low antibody titers with limited persistence. A third vaccine dose was safe and increased the antibody titers, suggesting that an increase in immunizations may be warranted in this population. PMID- 16362897 TI - Gene-expression profiling discriminates between cerebral malaria (CM)-susceptible mice and CM-resistant mice. AB - The development of cerebral malaria (CM) in mice with Plasmodium berghei ANKA infection is under genetic control. Brain gene-expression patterns were investigated in well-defined genetically CM-resistant (CM-R; BALB/c and DBA/2) and CM-susceptible (CM-S; C57BL/6 and CBA/J) mice by use of cDNA microarrays. By combining transcriptional profiling with rigorous statistical methods and cluster analysis, we identified a set of 69 genes that perfectly discriminated between mouse strains and between CM-R and CM-S mice. The analysis of gene ontological terms revealed that the genes that clustered and were related to susceptibility to CM preferentially belonged to some biological process classes, such as those pertaining to immune responses. Using a false discovery rate of 5% and the Welch t test, we identified 31 genes with consistent differential expression between CM R and CM-S mice. These data indicate that microarray analysis may be useful for identification of candidate genes that are potentially responsible for resistance or susceptibility to mouse CM and suggest that candidate genes identified in mice could be specifically tested in humans for an association with disease severity. PMID- 16362900 TI - Oral therapy for cholera: amino acids added to electrolyte solutions containing rice or glucose. PMID- 16362898 TI - T cells require tumor necrosis factor-alpha to provide protective immunity in mice infected with Histoplasma capsulatum. AB - We examined whether neutralization of tumor necrosis factor (TNF)-alpha after intranasal exposure of mice to Histoplasma capsulatum was necessary for control of primary or secondary infection. All mice given monoclonal antibody to TNF alpha on the day of infection or on day 3 after infection died. When antibody was administered on day 5 after infection, 60% of mice with primary infection died, whereas none with secondary infection did. Antibody treatment on day 7 after infection produced a transiently higher fungal burden. Because optimal clearance required TNF-alpha after the onset of infection, we hypothesized that it may regulate T cell function. Lung CD3+ cells were the dominant population of TNF alpha-producing cells (approximately 40%-70%). Neutralization of this cytokine decreased the number of memory T cells but not the number of activated, proliferating, or interferon-gamma-producing cells. T cells from infected, TNF alpha-neutralized mice failed to protect T cell-deficient mice. The absence of TNF-alpha induces a defect in T cell-mediated protection. PMID- 16362901 TI - Evaluating distance functions for clustering tandem repeats. AB - Tandem repeats are an important class of DNA repeats and much research has focused on their efficient identification, their use in DNA typing and fingerprinting, and their causative role in trinucleotide repeat diseases such as Huntington Disease, myotonic dystrophy, and Fragile-X mental retardation. We are interested in clustering tandem repeats into groups or families based on sequence similarity so that their biological importance may be further explored. To cluster tandem repeats we need a notion of pairwise distance which we obtain by alignment. In this paper we evaluate five distance functions used to produce those alignments--Consensus, Euclidean, Jensen-Shannon Divergence, Entropy Surface, and Entropy-weighted. It is important to analyze and compare these functions because the choice of distance metric forms the core of any clustering algorithm. We employ a novel method to compare alignments and thereby compare the distance functions themselves. We rank the distance functions based on the cluster validation techniques--Average Cluster Density and Average Silhouette Width. Finally, we propose a multi-phase clustering method which produces good quality clusters. In this study, we analyze clusters of tandem repeats from five sequences: Human Chromosomes 3, 5, 10 and X and C. elegans Chromosome III. PMID- 16362902 TI - Relationship between segmental duplications and repeat sequences in human chromosome 7. AB - Various types of repeat sequences are abundant in genomic sequences, and they are associated with the biological phenomena at distinct levels. In particular, comparative analyses of whole-genome-sized sequence data have revealed that repeat sequences cause segmental duplications, which are a type of chromosomal structural arrangement. In this study, we analyzed the relationships between segmental duplications and repeat sequences in human chromosome 7. For this purpose, three methods for detecting repeat sequences were applied to the genomic sequences of human chromosome 7: RepeatMasker for the dispersed repeats, TRF for the tandem repeats, and STEPSTONE for the inter-spread repeats. By plotting the detected repeat sequences against the locations on the chromosome, all three types of repeats were found to be concentrated around the regions of segmental duplications, as a macroscopic feature of their distributions. Furthermore, the latter two repeat sequences were classified in terms of their periods, and the distribution bias of the detected repeat sequences was statistically tested between the segmental duplication regions and the other regions. As a result, the periods of two repeats were biased, with less than a 5% level of significance probability by the chi(2) test, and the repeats with long periods, about 130bp and more than 400bp, were attributed to a bias with a 5% level of significance probability by the normalized residual test. The mechanism of segmental duplications is discussed based on the present results. PMID- 16362903 TI - Improvement in the accuracy of multiple sequence alignment program MAFFT. AB - In 2002, we developed and released a rapid multiple sequence alignment program MAFFT that was designed to handle a huge (up to approximately 5,000 sequences) and long data (approximately 2,000 aa or approximately 5,000 nt) in a reasonable time on a standard desktop PC. As for the accuracy, however, the previous versions (v.4 and lower) of MAFFT were outperformed by ProbCons and TCoffee v.2, both of which were released in 2004, in several benchmark tests. Here we report a recent extension of MAFFT that aims to improve the accuracy with as little cost of calculation time as possible. The extended version of MAFFT (v.5) has new iterative refinement options, G-INS-i and L-INS-i (collectively denoted as [GL] INS-i in this report). These options use a new objective function combining the weighted sum-of-pairs (WSP) score and a score similar to COFFEE derived from all pairwise alignments. We discuss the improvement in accuracy brought by this extension, mainly using two benchmark tests released very recently, BAliBASE v.3 (for protein alignments) and BRAliBASE (for RNA alignments). According to BAliBASE v.3, the overall average accuracy of L-INS-i was higher than those of other methods successively released in 2004, although the difference among the most accurate methods (ProbCons, TCoffee v.2 and new options of MAFFT) was small. The advantage in accuracy of [GL]-INS-i became greater for the alignments consisting of approximately 50-100 sequences. By utilizing this feature of MAFFT, we also examined another possible approach to improve the accuracy by incorporating homolog information collected from database. The [GL]-INS-i options are applicable to aligning up to approximately 200 sequences, although not applicable to thousands of sequences because of time and space complexities. PMID- 16362904 TI - A Gibbs sampling approach to detection of tree motifs. AB - This paper introduces a method to detect tree patterns (tree motifs) in a database of rooted unordered labeled trees. The method can be viewed as an extension of the Gibbs sampling approach to detect sequence motifs. Basically, we enumerate tree topologies and for each topology we seek within the database for tree motifs with the given topology. A tree motif can be detected by matching the tree topology against the database of trees and then applying Gibbs sampling on the matching set. After completion of the process for a given tree topology, the process is restarted for the next enumerated tree topology. The method outputs for each topology the best tree motif found. We applied our method to an artificially created database of trees as well as to a database of carbohydrate (glycan) structures. PMID- 16362905 TI - Antiviral activity of oligomerization-deficient Stat1. AB - Interferon stimulation of cells can activate several hundred target genes, many of which are required for antiviral protection. Promoter binding of tyrosine phosphorylated (activated) Stat1 dimers is essential for gene induction, a process that often entails the oligomerization of Stat1 dimers via interactions of their aminoterminal domains. The mutation of a single residue (F77) in the N domain of Stat1 was recently demonstrated to preclude both the dephosphorylation and the oligomerization of Stat1 dimers. Here, we investigated the influence of defective oligomerization on a complex phenotype such as the induction of an antiviral state. It was found that the antiviral protection conferred by interferon-a was strongly reduced, whereas the interferon-g response was not measurably affected. These results indicate that Stat1 oligomerization is required for the antiviral activity of interferons. Moreover, the concentration of activated Stat1 in the nucleus may generally play a critical role for interferon-induced target gene activation. PMID- 16362906 TI - Identification of conserved structural features at sequentially degenerate locations in transcription factor binding sites. AB - Many locations within transcription factor binding sites are not sequentially conserved and appear to be degenerate. We hypothesize that some of these positions contain essential structural codes that are recognized by the transcription factors that bind to them. The structural codes can be defined by base-pair step parameters that describe the relative displacement and orientation of two adjacent base pairs in a nucleic acid structure. We have developed a method, Identification of Conserved Structural Features (ICSF), which uses base pair step parameters obtained from a collection of high-resolution DNA crystal structures to discover structural conservation that exists in the sequentially degenerate areas within a binding site and produce profiles of the structural features along the entire site. We have focused our study on the transcription factor binding sites in the JASPAR database and have found that one-third (P value > or = 0.05) of the binding sites contain sequentially degenerate locations with highly conserved structural features as described by the base-pair step parameters. These results will help us to gain a better understanding of the process by which transcription factors recognize their binding sites and possibly lead to an improvement in our ability to find these sites in genomic sequences. AVAILABILITY: ICSF is freely available to academic users at http://zlab.bu.edu/ICSF . SUPPLEMENTARY INFORMATION: http://zlab.bu.edu/ICSF . PMID- 16362907 TI - Assessing transcription factor motif drift from noisy decoy sequences. AB - Genome scale identification of transcription factor binding sites (TFBS) is fundamental to understanding the complexities of mRNA expression at both the cell and organismal levels. While high-throughput experimental methods provide associations between transcription factors and the genes they regulate under a specified experimental condition, computational methods are still required to pinpoint the exact location of binding. Moreover, since the binding site is an intrinsic property of the promoter region, computational methods are in principle more general than condition dependent experimental methods. Computational identification of TFBSs is complicated in at least two different ways. First, transcription factors bind a heterogeneous distribution of sites and therefore have a distribution of affinities. Second, the set of sequences for which a common site is to be determined do not all have a site for the TF of interest. In this paper, we evaluate the robustness of TFBS identification with respect to both effects. We show addition of upstream regions that do not have the TFBS destroy the specificity of the predicted binding site. We also propose a method to calculate the distance between position weight matrices that can be used to measure "drift'' from the canonical binding site. The results presented here could be useful in developing future transcription factor binding site identification algorithms. PMID- 16362908 TI - Improvement of TRANSFAC matrices using multiple local alignment of transcription factor binding site sequences. AB - This paper describes a novel approach to constructing Position-Specific Weight Matrices (PWMs) based on the transcription factor binding site (TFBS) data provide by the TRANSFAC database and comparison of the newly generated PWMs with the original TRANSFAC matrices. Multiple local sequence alignment was performed on the TFBSs of each transcription factor. Several different alignment programs were tested and their matrices were compared to the original TRANSFAC matrices. One of the alignment programs, GLAM, produced comparable matrices in terms of the average ranking of true positive sites across the whole test set of sequences. PMID- 16362909 TI - On the kinetic design of transcription. AB - We analyse a stochastic model of transcription that describes transcription initiation by promoter activation and subsequent polymerase recruitment. Explicit expressions are derived for the control of an activator on the mean mRNA number and for the mRNA noise. Both properties are strongly influenced by the kinetics of promoter activation, mRNA synthesis and degradation. Low transcriptional noise is obtained either when the transcription initiation complex has a long life-time or when its components associate and dissociate rapidly. However, the ability of an activator to regulate the mRNA level is low in the first and high in the second case. Large noise is generated when the initial activation step of the promoter is slow. In this case, transcription can be burst-like; the mRNA distribution becomes bimodal while regulability of the mean copy number is maintained. PMID- 16362910 TI - Integrating genomic data to predict transcription factor binding. AB - Transcription factor binding sites (TFBS) in gene promoter regions are often predicted by using position specific scoring matrices (PSSMs), which summarize sequence patterns of experimentally determined TF binding sites. Although PSSMs are more reliable than simple consensus string matching in predicting a true binding site, they generally result in high numbers of false positive hits. This study attempts to reduce the number of false positive matches and generate new predictions by integrating various types of genomic data by two methods: a Bayesian allocation procedure, and support vector machine classification. Several methods will be explored to strengthen the prediction of a true TFBS in the Saccharomyces cerevisiae genome: binding site degeneracy, binding site conservation, phylogenetic profiling, TF binding site clustering, gene expression profiles, GO functional annotation, and k-mer counts in promoter regions. Binding site degeneracy (or redundancy) refers to the number of times a particular transcription factor's binding motif is discovered in the upstream region of a gene. Phylogenetic conservation takes into account the number of orthologous upstream regions in other genomes that contain a particular binding site. Phylogenetic profiling refers to the presence or absence of a gene across a large set of genomes. Binding site clusters are statistically significant clusters of TF binding sites detected by the algorithm ClusterBuster. Gene expression takes into account the idea that when the gene expression profiles of a transcription factor and a potential target gene are correlated, then it is more likely that the gene is a genuine target. Also, genes with highly correlated expression profiles are often regulated by the same TF(s). The GO annotation data takes advantage of the idea that common transcription targets often have related function. Finally, the distribution of the counts of all k-mers of length 4, 5, and 6 in gene's promoter region were examined as means to predict TF binding. In each case the data are compared to known true positives taken from ChIP-chip data, Transfac, and the Saccharomyces Genome Database. First, degeneracy, conservation, expression, and binding site clusters were examined independently and in combination via Bayesian allocation. Then, binding sites were predicted with a support vector machine (SVM) using all methods alone and in combination. The SVM works best when all genomic data are combined, but can also identify which methods contribute the most to accurate classification. On average, a support vector machine can classify binding sites with high sensitivity and an accuracy of almost 80%. PMID- 16362911 TI - Elucidation of the relationships between LexA-regulated genes in the SOS response. AB - Monitoring the expression of many genes under different conditions is a common approach for investigating gene relationships. In particular, the monitoring sheds light on the biological phenomena in which many genes are coordinately expressed. In this study, we analyzed the expression profiles of LexA-regulated genes after UV irradiation, to elucidate the genes related to the SOS response, which involves coordinately regulated gene expression. By the two-gene relationship analysis, the LexA-regulated genes were highly correlated with the genes involved in the DNA repair functions. The LexA-regulated genes with highly significant probability were divided into two groups: the LexA-regulated genes that were mutually related within them were related to the genes with DNA repair functions, while the LexA-regulated genes that were less related within them showed lower relation to the genes with DNA repair functions. By a multiple gene relationship analysis, the two types of LexA-regulated genes were clearly clustered, and the inferred network between the clusters indicated their sequential relationship of clusters in the two groups of LexA-regulated genes in the SOS response; the former type of genes emerged in the early stage of the SOS response upon the signal transduction by membrane proteins, cessation of cell division and recognition of DNA damage, and the latter type emerged in a later stage, and functioned in the repair mechanism and the resumption of DNA replication. PMID- 16362912 TI - Biological profiling of gene groups utilizing Gene Ontology. AB - Increasingly used high throughput experimental techniques, like DNA or protein microarrays give as a result groups of interesting, e.g. differentially regulated genes which require further biological interpretation. With the systematic functional annotation provided by the Gene Ontology the information required to automate the interpretation task is now accessible. However, the determination of statistical significance of a biological process within these groups is still an open question. In answering this question, multiple testing issues must be taken into account to avoid misleading results. Here we present a statistical framework that tests whether functions, processes or locations described in the Gene Ontology are significantly enriched within a group of interesting genes when compared to a reference group. First we define an exact analytical expression for the expected number of false positives that allows us to calculate adjusted p values to control the false discovery rate. Next, we demonstrate and discuss the capabilities of our approach using publicly available microarray data on cell cycle regulated genes. Further, we analyze the robustness of our framework with respect to the exact gene group composition and compare the performance with earlier approaches. The software package GOSSIP implements our method and is made freely available at http://gossip.gene-groups.net/. PMID- 16362913 TI - Conservation of gene co-regulation between two prokaryotes: Bacillus subtilis and Escherichia coli. AB - We measured conservation of gene co-regulation between two distantly related prokaryotes, B. subtilis and E. coli. The co-regulation between genes was extracted from knowledge of regulation of genes stored in databases. For B. subtilis operons, we obtained the data set from ODB which we have developed and, for the regulons, we used DBTBS. For E. coli data set, we used known regulons derived from RegulonDB. We obtained a reliable data set of co-regulated genes in B. subtilis and E. coli. About 60-80 % of gene pairs conserved co-regulation relationships, so co-regulation between genes are highly conserved even between distantly related species. To measure the functional relationship between these conserved genes, we used KEGG PATHWAY and COG. When two co-regulated genes are in the same biological pathway in KEGG or share the same functional category in COG, we assume that they have the same function. As a result, we also found that many conserved co-regulated gene pairs share the same functions. These observations would help to predict gene co-regulation and protein functions. PMID- 16362914 TI - Comparative analysis of cell cycle regulated genes in eukaryotes. AB - We compared microarray experiments on cell cycle of three model eukaryotes: budding and fission yeast and human cells. Only 112 orthologous groups were cyclic in the three model organisms. The common set of cyclic orthologs includes many taking part in the cell cycle progression, like cyclin B homologs, CDC5, SCH9, DSK2, ZPR1. Proteins involved in DNA replication included histones, some checkpoint kinases and some proteins regulating DNA damage and repair. Conserved cyclic proteins involved in cytokinesis included myosins and kinesins. Many groups of genes related to translation and other metabolic processes were also cyclic in all three organisms. This reflects rebuilding of cellular components after the replication and changes of metabolism during the cell cycle. Many genes important in cell cycle control are not cyclic or not conserved. This includes transcription factors implicated in the regulation of budding yeast cell cycle. The partially overlapping roles of regulatory proteins might allow the evolutionary substitution of components of cell cycle. PMID- 16362915 TI - Efficient determination of cluster boundaries for analysis of gene expression profile data using hierarchical clustering and wavelet transform. AB - The existing methods for clustering of gene expression profile data either require manual inspection and other biological knowledge or require some cut-off value which can not be directly calculated from the given data set. Thus, the problem of systematic and efficient determination of cluster boundaries of clusters in gene expression profile data still remains demanding. In this context, we have developed a procedure for automatic and systematic determination of the boundaries of clusters in the hierarchical clustering of gene expression data based on the ratio of with-in class variance and between-class variance, which can be fully calculated from the given expression data. After the determination of dendrogram based on agglomerative hierarchical clustering, this ratio is used to determine the cluster boundary. Except this ratio which can be completely calculated from the given expression profile data, unlike other existing approaches, our approach does not require any manual inspection or biological knowledge. Our results are favorably comparable and in some of cases better than existing method which does not utilize prior information or manual inspection. Moreover, gene expression profile data are often contaminated with various type of noise and in order to reduce this noise content, we have also applied image enhancing technique called discrete wavelet transform. We tested a number of mother wavelet functions to smooth the noise in the gene expression data set and obtained some improvements in the quality of the results. PMID- 16362916 TI - Deciphering protein network organization using phylogenetic profile groups. AB - Phylogenetic profiling is now an effective computational method to detect functional associations between proteins. The method links two proteins in accordance with the similarity of their phyletic distributions across a set of genomes. While pair-wise linkage is useful, it misses correlations in higher order groups: triplets, quadruplets, and so on. Here we assess the probability of observing co-occurrence patterns of 3 binary profiles by chance and show that this probability is asymptotically the same as the mutual information in three profiles. We demonstrate the utility of the probability and the mutual information metrics in detecting overly represented triplets of orthologous proteins which could not be detected using pairwise profiles. These triplets serve as small building blocks, i.e. motifs in protein networks; they allow us to infer the function of uncharacterized members, and facilitate analysis of the local structure and global organization of the protein network. Our method is extendable to N-component clusters, and therefore serves as a general tool for high order protein function annotation. PMID- 16362917 TI - Comprehensive analysis and prediction of synthetic lethality using subcellular locations. AB - The lethality of a gene is a fundamental and representative measure for understanding the function of a gene and its associated bio-systems. Recently, many research groups have started focusing on the concept of synthetic lethality. The synthetic lethality between genes is defined by the combination of mutations in two genes causing cell death. Here, we confirm that synthetic lethality and cellular location have close relationships among the Saccharomyces cerevisiae genes. Furthermore, we attempt the prediction of candidate gene pairs with synthetic lethality. The prediction is based on the hierarchical aspect model (HAM) which learns from a data set of cellular location to estimate a likelihood value indicating the synthetic lethality between genes. PMID- 16362918 TI - Lethality and entropy of protein interaction networks. AB - We characterize protein interaction networks in terms of network entropy. This approach suggests a ranking principle, which strongly correlates with elements of functional importance, such as lethal proteins. Our combined analysis of protein interaction networks and functional profiles in single cellular yeast and multi cellular worm shows that proteins with large contribution to network entropy are preferentially lethal. While entropy is inherently a dynamical concept, the present analysis incorporates only structural information. Our result therefore highlights the importance of topological features, which appear as correlates of an underlying dynamical property, and which in turn determine functional traits. We argue that network entropy is a natural extension of previously studied observables, such as pathway multiplicity and centrality. It is also applicable to networks in which the processes can be quantified and therefore serves as a link to study questions of structural and dynamical robustness in a unified way. PMID- 16362919 TI - Structural and dynamical analyses of the kinase network derived from the transpath database. AB - We analyze the structural design and the dynamical properties of a protein kinase network derived from the Transpath database. We consider structural properties, such as feedback cycles, pathway lengths, fraction of shortest pathways and crosstalk. Dynamic characteristics of the network are analyzed by using nonlinear differential equations with a special focus on kinase amplitudes and signal propagation times. Comparison with random networks shows that the cellular kinase network exhibits special features which might be a result of natural selection. In particular, the Transpath network contains no cycles, and input kinases and output kinases are generally connected by shortest signalling routes. Moreover, it displays a characteristic spectrum of cross-talk between different pathways. PMID- 16362920 TI - Comparative analysis of DNA-binding proteins between thermophilic and mesophilic bacteria. AB - Thermophilic bacteria are one of the most attractive forms of life, and their adaptation mechanisms to elevated temperatures have been extensively studied over the years. Thermal adaptations of cell components such as proteins and RNA are well studied, but adaptations of interactions between these components must be also vital for the thermophiles. Protein-DNA interactions play crucial roles in the cell, but little is known about their thermal adaptations. In this study, we analyzed DNA-binding proteins from thermophilic bacteria. Comparison of amino acid compositions at the DNA-binding interfaces between thermophiles and their mesophilic close relatives revealed several commonalities between phylogenetically unrelated organisms. Advantages and limitations of our methods will be also discussed. PMID- 16362921 TI - Identifying drug active pathways from gene networks estimated by gene expression data. AB - We present a computational method for identifying genes and their regulatory pathways influenced by a drug, using microarray gene expression data collected by single gene disruptions and drug responses. The automatic identification of such genes and pathways in organisms' cells is an important problem for pharmacogenomics and the tailor-made medication. Our method estimates regulatory relationships between genes as a gene network from microarray data of gene disruptions with a Bayesian network model, then identifies the drug affected genes and their regulatory pathways on the estimated network with time course drug response microarray data. Compared to the existing method, our proposed method can identify not only the drug affected genes and the druggable genes, but also the drug responses of the pathways. For evaluating the proposed method, we conducted simulated examples based on artificial networks and expression data. Our method succeeded in identifying the pseudo drug affected genes and pathways with the high coverage greater than 80 %. We also applied our method to Saccharomyces cerevisiae drug response microarray data. In this real example, we identified the genes and the pathways that are potentially influenced by a drug. These computational experiments indicate that our method successfully identifies the drug-activated genes and pathways, and is capable of predicting undesirable side effects of the drug, identifying novel drug target genes, and understanding the unknown mechanisms of the drug. PMID- 16362922 TI - Prediction of debacle points for robustness of biological pathways by using recurrent neural networks. AB - Living organisms have ingenious control mechanisms in which many molecular interactions work for keeping their normal activities against disturbances inside and outside of them. However, at the same time, the control mechanism has debacle points at which the stability can be broken easily. This paper proposes a new method which uses recurrent neural network for predicting debacle points in an hybrid functional Petri net model of a biological pathway. Evaluation on an apoptosis signaling pathway indicates that the rates of 96.5 % of debacle points and 65.5 % of non-debacle points can be predicted by the proposed method. PMID- 16362923 TI - A cross species comparison of metabolic network functions. AB - We compare a large number of organisms with respect to their metabolic network functions. We measure such functions in terms of the synthesizing capacity of a network when it is provided with a few small chemical substances as external resources. We call this measure the scope and show that it is generally robust against structural alterations of the reaction network. Organisms can be separated into two groups, one with a small and one with a large scope. Networks with a high synthesizing capacity also show a high degree of robustness against structural changes, indicating that this network function has been a target in the evolutionary past of the corresponding organisms. A comparison between structural and functional similarities reveals that organisms with a similar structure do not necessarily show similar biological functions. The presented concepts allow for a systematic investigation of structure-function relationships of metabolic networks and may put forth valuable hints on the evolution of metabolic pathways. PMID- 16362924 TI - A global representation of the carbohydrate structures: a tool for the analysis of glycan. AB - Glycan resources have been developed of late, such as carbohydrate databases, analysis tools, and algorithms for analysis of carbohydrate features. With this background, bioinformatics approaches to carbohydrate research have recently begun using a large amount of protein and carbohydrate data. This paper introduces one of these projects that elucidates the range of carbohydrate structures. In this study, the variety of carbohydrate structures have been enumerated in a global tree structure called variation trees, using the KEGG GLYCAN database, which is a public-domain glycan resource for bioinformatics analysis. Additionally, a glycosyltransferase mapping list of glycosyltransferases and their catalyzing glycosidic linkages was constructed. From this, we present the composite structure map (CSM), which is a structural variation map integrating its variation trees and glycosyltransferase map list. CSM is able to display, for example, expression data of glycosyltransferases in a compact manner, illustrating its versatility as a new bioinformatics resource and tool capable of analyzing carbohydrate structures on a global scale. These resources are available at http://www.genome.jp/kegg/glycan/. PMID- 16362925 TI - Combining bioinformatics resources for the structural modelling of eukaryotic metabolic networks. AB - The architecture of the cellular metabolic network is almost completely available from several databases. This has paved the way for computational analyses. Whereas kinetic modelling is still restrained to small metabolic sub-systems for which enzyme-kinetic details are known, so-called structural modelling techniques can be applied to complete metabolic networks even if the kinetics and regulation of the underlying enzymes is still unknown. Structural modelling requires detailed information on the presence of metabolic enzymes in a specific cell type of interest and the thermodynamics of the reactions, determining their direction under cellular conditions. If compartments are distinguished the sub-cellular compartmentation of reactions and enzymes and the membrane transporters exchanging metabolites between cellular compartments must be included. All this information cannot be taken from a single data base but has to be compiled from various Bioinformatics resources. Here we present an approach towards the organization of Bioinformatics data that enables the flux-balance analysis of comprehensive compartmentalized metabolic networks of eukaryotic cells with special focus on human hepatocytes. PMID- 16362926 TI - Robust diagnosis of non-Hodgkin lymphoma phenotypes validated on gene expression data from different laboratories. AB - A major challenge in cancer diagnosis from microarray data is the need for robust, accurate, classification models which are independent of the analysis techniques used and can combine data from different laboratories. We propose such a classification scheme originally developed for phenotype identification from mass spectrometry data. The method uses a robust multivariate gene selection procedure and combines the results of several machine learning tools trained on raw and pattern data to produce an accurate meta-classifier. We illustrate and validate our method by applying it to gene expression datasets: the oligonucleotide HuGeneFL microarray dataset of Shipp et al. (www.genome.wi.mit.du/MPR/lymphoma) and the Hu95Av2 Affymetrix dataset (DallaFavera's laboratory, Columbia University). Our pattern-based meta classification technique achieves higher predictive accuracies than each of the individual classifiers , is robust against data perturbations and provides subsets of related predictive genes. Our techniques predict that combinations of some genes in the p53 pathway are highly predictive of phenotype. In particular, we find that in 80% of DLBCL cases the mRNA level of at least one of the three genes p53, PLK1 and CDK2 is elevated, while in 80% of FL cases, the mRNA level of at most one of them is elevated. PMID- 16362927 TI - Simple discriminant functions identify small sets of genes that distinguish cancer phenotype from normal. AB - High-throughput gene expression profiling can identify sets of genes that are differentially expressed between different phenotypes. Discovering marker genes is particularly important in diagnosis of a cancer phenotype. However, gene sets produced to date are too large to be economically viable diagnostics. We use a hybrid decision tree-discriminant analysis to identify small sets of genes, i.e. single genes and gene pairs, which separate normal samples from different stages of tumor samples. Half the samples are selected for training to form the probability distribution of expression values of each gene. The distributions for the tumor and normal phenotypes are then used to classify the test samples. The algorithm also identifies gene pairs by combining the probability distributions to construct a decision tree which is used to determine the class of test samples. After a series of training and testing sessions, genes and gene pairs that classify all samples correctly are recorded. The method was applied to a breast cancer data; and classifier genes that distinguish normal breast from different stages of breast tumor were identified. The genes were ranked according to their minimum Euclidean distance between the expression values in tumor and normal samples. The algorithm was able to pick known cancer related genes but also find genes that were not identified as differentially expressed by t-test with a 2 fold cut-off. Overall, the method generates possible diagnostic genes and gene pairs for a specific disease phenotype to pursue further biological interpretations in cancer biology. PMID- 16362928 TI - Selecting stable molecular targets for treatment and prevention of AIDS. AB - The common consideration in approaching protection against HIV, whether by a vaccine or therapeutic, is identification of suitable targets. Among the central criteria for suitability is target stability; i.e. resistance to mutation. In this paper we address the problem of stability, and develop methods for identifying stable targets. The targets that we focus on are structures formed by viral peptides and products of the class I major histocompatibility complex, the target of the immune system. The method mines the large databases of fully sequenced HIV genomes and MHC binding peptides, and takes account of human polymorphism to construct hundreds of subpopulation specific stable targets, each consisting of combinations of 3-5 complexes. PMID- 16362929 TI - Aggregation in Huntington's disease: insights through modelling. AB - Huntington's Disease (HD) is a late-onset, progressively degenerative brain disorder characterized by cell loss in the striatum and cortex. HD is caused by a polyglutamine (polyQ) expansion in the protein huntingtin (Htt). The mutant Htt is a substrate for caspases -2, -3 and -6. The cleavage of mutant Htt by caspase 2 has been suggested to underlie the selective neuronal death in HD. Once the mutant Htt is cleaved, a sticky and toxic fragment with the potential to form aggregates is released. The role of aggregation in the progression of HD has been extensively studied, yielding a plethora of ambivalent results. It has been shown that these are the diffuse, monomeric and oligomeric, forms of the mutant Htt fragment rather than the aggregates that are the major source of toxicity to the cells. We present here a mathematical model for aggregation in HD and discuss how it can relate to the selective neuronal death and the dependence of the disease onset on polyQ length. We describe the dynamical behavior of caspase-2, the release of monomeric forms of the mutant Htt fragment and the aggregation of these fragments through intermediate steps. Our model predicts that the concentration of toxic, intermediate oligomeric structures does not increase with increased caspase activity. We therefore suggest that the intermediate oligomeric forms of toxic Htt fragment do not account for selective and polyQ dependent neuronal death. PMID- 16362930 TI - Autoimmune diseases and peptide variations. AB - The immune system plays an essential role in the defense of the host against invaders. Enormous numbers of lymphocytes are recruited and immense numbers of antibodies or cytokines are secreted in various kinds of immune response. But the system also has the possibility of being the cause of tissue injury or some kind of diseases. For example, when their functions target their host, an autoimmune disease occurs. Although the pathogenesis of various autoimmune diseases has been scrutinized intensively, there is little evidence as of yet. But it has been reported that in most of the disease subjects, a broad spectrum of antibodies recognizing components of self tissue or circulating self antigens that normally should be ignored are observed. In this study, we come to the conclusion that proteins targeted by these autoreactive antibodies share the same peptides with some kind of proteins of viruses known to infect human. This result supports the fact that viral infection is a speculative cause of the disease in some subjects. PMID- 16362932 TI - PII structure in the model peptides for unfolded proteins: studies on ubiquitin fragments and several alanine-rich peptides containing QQQ, SSS, FFF, and VVV. AB - A great deal of attention has been paid lately to the structures in unfolded proteins due to the recent discovery of many biologically functional but natively unfolded proteins and the far-reaching implications of order in unfolded states for protein folding. Recently, studies on oligo-Ala, oligo-Lys, oligo-Asp, and oligo-Glu, as well as oligo-Pro, have indicated that the left-handed polyproline II (PII) is the major local structure in these short peptides. Here, we show by NMR and CD studies that ubiquitin fragments, model unfolded peptides composed of nonrepeating amino acids, and four alanine-rich peptides containing QQQ, SSS, FFF, and VVV sequences are all present in aqueous solution predominantly in the extended PII or beta conformation. The results from this and related studies indicate that PII might be a major backbone conformation in unfolded proteins. The presence of defined local backbone structure in unfolded proteins is inconsistent with predictions from random coil models. PMID- 16362933 TI - Folding mechanism of beta-hairpins studied by replica exchange molecular simulations. AB - The folding process of trpzip2 beta-hairpin is studied by the replica exchange molecular dynamics (REMD) and normal MD simulations, aiming to understand the folding mechanism of this unique small, stable, and fast folder, as well as to reveal the general principles in the folding of beta-hairpins. According to our simulations, the TS ensemble is mainly characterized by a largely formed turn and the interaction between the inner pair of hydrophobic core residues. The folding is a zipping up of hydrogen bonds. However, the nascent turn has to be stabilized by the partially formed hydrophobic core to cross the TS. Thus our folding picture is in essence a blend of hydrogen bond-centric and hydrophobic core centric mechanism. Our simulations provide a direct evidence for a very recent experiment (Du et al., Proc Natl Acad Sci USA 2004;101:15915-15920), which suggests that the turn formation is the rate-limiting step for beta-hairpin folding and the unfolding is mainly determined by the hydrophobic interactions. Besides, the relationship between hydrogen bond stabilities and their relative importance in folding are investigated. It is found that the hydrogen bonds with higher stabilities need not play more important roles in the folding process, and vice versa. PMID- 16362935 TI - Adaptations to humeral torsion in medieval Britain. AB - Adaptations to the humeral torsion angle have been identified in the professional throwing athlete. This movement pattern increases the humeral torsion angle, and also increases the extent of external rotation movements in the dominant, throwing limb when compared with the nondominant limb. The purpose of this paper is to test the hypothesis that the humeral torsion angle is an adaptation to upper limb use. This project examines the humeral torsion angle in a number of medieval British populations, as well as a modern cadaver-based sample. The results identify significant differences in the humeral torsion angle both between and within male (P < 0.001, ANOVA) and female (P < 0.014, ANOVA) populations, although the results are not consistent with expected behavior patterns. Statistically significant differences between males and females within the same site were identified in 2 of the 5 samples examined. The mean level of bilateral asymmetry does not approach that reported for the professional throwing athlete. However, a number of individuals have high levels of asymmetry in excess of that identified in the professional throwing athlete. This analysis demonstrates the need for individual rather than population-based analyses, as the heterogeneity within population samples obscures individual variation in activity patterns. The diversity within British medieval society and a lack of specific known behaviors prevent further identification of the functional significance of the humeral torsion angle within the archaeological record examined here. PMID- 16362936 TI - Insights on HIV-1 Tat:P/CAF bromodomain molecular recognition from in vivo experiments and molecular dynamics simulations. AB - Structural and functional studies indicate that, through its bromodomain, the cellular acetyltransferase P/CAF binds the acetylated Tat protein of human immunodeficiency virus type 1 (HIV-1) and promotes transcriptional activation of the integrated provirus. Based on the NMR structure of P/CAF complexed with an acetylated Tat peptide, here we use molecular dynamics simulations to construct a model describing the interaction between full length Tat and the P/CAF bromodomain. Our calculations show that the protein-protein interface involves hydrophobic interactions between the P/CAF ZA loop and the Tat core domain. In particular, tyrosines 760 and 761 of P/CAF, two residues that are highly conserved in most known bromodomains, play an essential role for the binding. Fluorescence resonance energy transfer (FRET) experiments performed in this work demonstrate that P/CAF proteins in which these tyrosines are mutated into hydrophilic residues neither bind to Tat inside the cells nor mediate Tat transactivation. The combination of theoretical and in vivo studies provides new insights into the specificity of bromodomain recognition. PMID- 16362937 TI - The prevalence of and risk factors for back pain among home care nursing personnel in Hong Kong. AB - BACKGROUND: There is a large gap in research with regard to back pain (BP) among home care nursing personnel (HCNP); only seven studies have been conducted worldwide. There is a need to identify the magnitude of and risk factors for BP that are unique to Hong Kong (HK) HCNP. METHODS: A total population sampling technique was employed in this cross-sectional questionnaire-based study. Hierarchical multiple logistic regression analyses were used to control for potentially confounding variables. RESULTS: The 12-month prevalence of upper and lower BP was 71.2% (n=265). Three predictors were identified: physical risk factors in the office (OR=3.57, 95% CI=1.55-8.24), static postures (OR=1.41, 95% CI=1.04-1.90), and psychological job demands (OR=1.11, 95% CI=1.01-1.22). CONCLUSION: HCNP in HK have a high prevalence of BP. BP in HK HCNP is independently attributable to physical work factors in the office, static postures, and psychological job demands, and is not primarily associated with patient lifting and transferring which are traditionally identified as risk factors for BP in hospital nursing personnel. Am. J. Ind. Med. 49:14-22, 2006. (c) 2005 Wiley-Liss, Inc. PMID- 16362938 TI - Policemen and firefighters have increased risk for type-2 diabetes mellitus probably due to their large body mass index: a follow-up study in Japanese men. AB - BACKGROUND: Policemen/firefighters may have increased risk for mortality from ischemic heart disease and from all causes. We compare incidence of type-2 diabetes mellitus (DM), a well-known predictor for mortality, in policemen/firefighters with that in two other worker-groups. METHODS: A follow-up study of 5,130 healthy Japanese men aged 30-49 years at baseline consisted of three worker-groups; 3,111 clerical workers, 1,122 manual/production and transport/communication workers, and 897 policemen/firefighters. Incident DM was identified by "fasting serum glucose >or=7.00 mmol/L" or/and "under medical treatment for DM." Hazard ratio (HR) and 95% confidence interval (95% CI) for incident DM was estimated by Cox's proportional hazard models using clerical workers as a reference group (HR=1.00). Baseline age, body mass index (BMI, kg/m(2)), drinking, smoking, exercise, and education were computed as confounders. RESULTS: During mean follow-up periods of 8.4 years, 155 clerical workers, 51 manual/production and transport/communication workers, and 74 policemen/firefighters had incident DM (5.9, 5.7, and 9.6 per 1,000 person-years, respectively; P=0.001 by chi(2)-test). Policemen/firefighters had significantly increased risk for incident DM against clerical workers (age-adjusted HR (95% CI): 1.65 (1.25, 2.18)), but the significance disappeared after adjustments for BMI (age- and BMI-adjusted HR (95% CI): 1.16 (0.87, 1.54)). Manual/production and transport/communication workers had no increased risk. CONCLUSIONS: Policemen/firefighters have increased risk for DM probably due to their large BMI. Body weight control may be more efficient for prevention of DM and DM related health problems in policemen/firefighters than in other workers. PMID- 16362939 TI - The BeLPT: algorithms and implications. AB - BACKGROUND: The beryllium lymphocyte proliferation test (BeLPT) is used to identify persons with beryllium sensitization. The variability of laboratory results and lack of a "gold standard" have led to questions about the test's performance. Fortunately, a recently published study has credibly estimated standard epidemiologic parameters for the BeLPT. METHODS: Information from this recent study was used to assess the performance of two common algorithms for BeLPT testing. Standard epidemiologic parameters were determined for two common algorithms and then compared. RESULTS: One of the two algorithms was more sensitive than the other (86% vs. 66%). The specificity of both algorithms (99.8% or greater) was high. At an estimated 2% prevalence, the positive predictive value of both algorithms for beryllium sensitization remained high (90% or higher). CONCLUSIONS: A priori characterization of the testing algorithm under consideration can enhance public health decision-making. PMID- 16362940 TI - Assessment of occupational eye injury risk and severity: an analysis of Rhode Island workers' compensation data 1998-2002. AB - BACKGROUND: Occupational eye injuries have been recognized as a serious health risk to workers and are in need of further investigation to develop effective interventions. METHODS: Rhode Island workers' compensation claims of ocular injury between 1998 through 2002 (n=8,877) were examined. The Current Population Survey was used to estimate occupational employment levels as a baseline for rate calculations. RESULTS: The estimated ocular injury claim rate was 32.9 per 10,000 workers (95% CI=32.3-33.6), with the cost of claims totaling $1,514,666 and averaging $171 per claim. The highest estimated claim rate of all occupations was found for construction laborers of 373.7 per 10,000 workers (95% CI=267.1-480.3). Relative to the durable manufacturing industry, the highest risk of injury resulting in disability indemnification was the wholesale trade industry (OR=2.18, 95% CI=1.19-4.01, P<0.05). CONCLUSIONS: Many of the eye injuries reported were likely preventable. Greater diligence, training, and safety precautions are needed to reduce the risk of eye injury to employees. PMID- 16362941 TI - Re: Evaluation of the size and type of free particulates collected from unused asbestos-containing brake components as related to potential for respirability. AB - BACKGROUND: In Atkinson et al. 2004 rinsates of unused brake components were analyzed by transmission electron microscopy (TEM) for the presence of asbestos fibers. RESULTS: We do not believe that the findings of Atkinson et al. are informative and could have been predicted based on the study design and the fact that one would expect to find measurable TEM asbestos fibers on an unused brake component. We also find that the paper did not provide a full or even partial discussion of the published literature with respect to industrial hygiene or epidemiology data. CONCLUSION: The findings of Atkinson et al. do not, in our view, "further raise concerns" about historical asbestos exposures experienced by automotive mechanics because of the vast amount of published literature to the contrary. PMID- 16362942 TI - Estimation of future mortality from pleural malignant mesothelioma in Japan based on an age-cohort model. AB - BACKGROUND: Japanese consumption of asbestos increased rapidly after the 1950s and lingered at a high level while the world's consumption decreased substantially after the 1980s. Mesothelioma is due primarily to asbestos, and the number of deaths in Japan is expected to increase in the future. METHOD: We estimated the future number of pleural mesothelioma deaths among males in Japan using an age-cohort model. RESULTS: Analyses showed that there would be about 100,000 deaths in Japan due to pleural mesothelioma in the next 40 years. Compared with the statistics in European countries, the ratio of expected death numbers to the population size is remarkably close to linear. The data-point for Japan was slightly lower than that which could be expected from the linear relationship. CONCLUSIONS: The limited availability of data may result in underestimation. Taking into consideration the consumption pattern of asbestos in recent decades, the incorporation of later cohorts will improve the estimation. PMID- 16362943 TI - Variations in diagnostic criteria for carpal tunnel syndrome among Ontario specialists. AB - BACKGROUND: Variations in diagnostic criteria for carpal tunnel syndrome (CTS) may result in differing reports of disease prevalence, errors in diagnosis, and variable results of treatment. The objective of this study was to determine how consistent specialists are in their ratings of the importance of clinical criteria for the diagnosis of CTS. METHODS: Three hundred specialist physicians and surgeons received a questionnaire containing 57 clinical criteria for the diagnosis of CTS. A visual analog scale (VAS) was used to rate the importance of each criterion in the diagnosis of CTS. RESULTS: The overall consistency both across and within specialties was poor (intraclass correlation coefficient across specialties (ICC) = 0.28; ICC range within specialties 0.27-0.37). CONCLUSIONS: Specialists are relatively inconsistent in the importance they assign to clinical criteria for the diagnosis of CTS. This inconsistency may be an important source of variation in the reported prevalence and treatment of CTS. PMID- 16362945 TI - Laboratory evaluation of belt usage with apple buckets. AB - BACKGROUND: Migrant and seasonal orchard harvest workers experience musculoskeletal strain caused by carrying heavy loads and holding awkward postures. An ergonomic hip belt designed to redistribute weight from the upper back, neck, and shoulders to the hips was evaluated. METHODS: A comparison of muscle exertion (using electromyography [EMG]) of four key back and shoulder muscles and shoulder surface pressure (using a surface pressure sensor [PS]) was made among 10 laboratory volunteers under intervention and control conditions, in two postures (standing erect [0 degrees ]) and flexed [45 degrees ]). RESULTS: All but one measure showed a significant reduction in the flexed posture with the intervention. Two of eight measures showed a significant reduction in the erect posture. Correlations with shoulder discomfort were found for deep shoulder EMG (r=0.71, P=0.021) and shoulder surface pressure (r=0.58, P=0.082). CONCLUSIONS: The hip belt intervention produces significant reductions in muscle activity while in a posture commonly assumed during orchard work. PMID- 16362946 TI - The last Senior Medical Inspector of Factories and his place in the history of occupational health. AB - BACKGROUND: Apart from a few industrialists with senses of enlightened self interest and of ethics, 19th century Occupational Health (OH) in the United Kingdom was essentially in the care of Her Majesty's Factory Inspectorate (HMFI). Two World Wars drew attention to the special health and efficiency needs of workers in the armaments industry, but it required the climate of a Welfare State after the Second for more general provision to be considered. A number of committees made recommendations for enhancing OH that were not implemented, and it was not until 1972 that a political will developed to enhance the State's provision. As a result of a far-sighted senior civil servant, a remarkable Senior Medical Inspector was in post, ready and willing to assist in bringing about change. METHODS: Published official materials have been reviewed in the light of the author's observation of events and persons from 1967 onwards. RESULTS: For a few years, as a consequence of a political will, academic, corporate, and governmental OH burgeoned in Britain, but with a decline in the economy and political change favoring deregulation, it underwent regression. CONCLUSIONS: Any number of committees may meet to discuss OH provision but their reports will moulder until there is a will for implementing their recommendations, at which the requisite funds and persons materialize to establish institutions. When the political climate changes, it will not be difficult to find persons who for a consideration will assist in their demolition. PMID- 16362948 TI - Developing search strategies in Medline on the occupational origin of diseases. AB - BACKGROUND: In order to support occupational health practitioners, researchers, and experts (clinical librarians, guideline developers) who search the scientific literature in PubMed (Medline), we developed and evaluated an easy-to-use and effective search strategy. METHODS: We tested different combinations of search terms, in answering four case vignettes calculating sensitivity and precision in retrieving literature from a gold standard. The use of the Clinical Queries filter from PubMed was evaluated. Based on the outcomes, we developed guidelines for a professional search strategy, using a flow chart. RESULTS: For the occupational health practitioner, we found a sensitivity of at least 65% with a precision of 20%, when terms for occupation and type of industry were combined with terms for disease. For the occupational health expert, a high sensitivity of more than 90% was found adding terms for risk factors to the strategy. Combining the search terms with the sensitive Clinical Queries etiology filter in PubMed, enhanced the precision. Using the specific Clinical Queries etiology filter in PubMed results in a substantial loss of relevant articles. CONCLUSIONS: To search for studies on the occupational origin of diseases in Medline, we propose to start with the proper name for the occupational disease. If this does not exist, use a specific term for the occupation and type of industry in combination with terms for disease. To improve the sensitivity of the search, a search term for the specific risk factor should be added. If there is no relevant occupational title available, it is worth trying the general search terms occupational diseases or occupational risk. PMID- 16362949 TI - Mortality in Florida professional firefighters, 1972-1999. PMID- 16362950 TI - Exposures to silica mixed dust and cohort mortality study in tin mines: exposure response analysis and risk assessment of lung cancer. AB - BACKGROUND: Mineral dusts that contain crystalline silica have been associated directly or indirectly with the development of pneumoconiosis or silicosis, non malignant respiratory diseases, lung cancer, and other diseases. The health impacts on workers with silica mixed dust exposure in tin mines and dose-response relationships between cumulative dust exposure and the mortality from lung cancer are investigated. METHODS: A cohort of 7,837 workers registered in the employment records in 4 Chinese tin mines between 1972 and 1974 was identified for this study and the mortality follow-up was traced through 1994. Of the cohort, the cause of death was ascertained for 1,061 (97%) of the 1,094 deceased workers. Standardized mortality ratios (SMRs) were calculated for all workers, non-exposed workers, and dust-exposed workers with different exposure levels, silicotics, and non-silicotics based on Chinese national rates. RESULTS: The mortality from all causes in four tin mines was nearly the same as the national mortality. Malignant neoplasm, cerebrovascular disease, and cardiovascular disease accounted for 68.6% of all deaths. Mortality excess from lung cancer, liver cancer, all malignant diseases, and non-malignant respiratory diseases was observed among dust-exposed workers; a 50-fold excess of pneumoconiosis was observed. There was an upward trend for SMRs of lung cancer was noted from no exposure to low, medium, and high exposure levels (SMRs=1.29, 2.65, 2.66, 3.33). The shape of the exposure-response curve for risk of lung cancer at high exposure levels was inconsistent in these four mines. CONCLUSIONS: The findings indicated a positive dose-response relation between exposure to cumulative dust and the mortality of lung cancer. High arsenic content in dust particles, together with crystalline silica, may play an important role in causing increased mortality from lung cancer. PMID- 16362951 TI - Workplace air quality and lung function among dental laboratory technicians. AB - BACKGROUND: Dental laboratory technicians are exposed to dust and/or methyl methacrylate (MMA) in the workplace. Potential effects of the exposures on lung function merit further clarification. METHODS: This study measured lung function of 45 dental technicians, before and after shifts, on 3 workdays and monitored workplace concentrations of particulate matter with aerodynamic diameter less than 2.5 mm (PM2.5) and MMA in five dental laboratories in Taiwan. Base metals in particles were analyzed by X-ray fluorescence. Repeated measurement analysis was applied to estimate potential effects of PM2.5 on pulmonary function. RESULTS: PM2.5 ranged from 26 microg/m3 to 664 microg/m3 and levels of MMA were low in these laboratories. Nickel, cobalt, and chromium were detected in the particles. After controlling for important factors, PM2.5 was associated with a small decrease in forced vital capacity and forced expiratory flow in one second. CONCLUSION: Workplace PM2.5 was associated with a non-significant decrease in lung function of dental technicians. PMID- 16362952 TI - Skin symptoms and work-related skin symptoms among grape farmers in Crete, Greece. AB - BACKGROUND: Grape farmers are exposed to a variety of agents capable of inducing occupational skin disease. We conducted a study to measure the prevalence of skin symptoms and work-related skin symptoms among grape farmers in the Malevisi region of Crete and to provide data on associated risk factors. METHODS: One hundred twenty grape farmers and 100 controls participated in the study. The protocol consisted of a questionnaire, skin prick tests for 16 common allergens, and measurement of specific IgE antibodies against 8 allergens. RESULTS: Self reported itchy rash (OR, 2.31; 95%CI, 1.10-4.84, P<0.05) within the last 12 months, and work-related itchy rash (OR, 4.08; 95%CI, 1.01-20.33, P<0.05) were significantly higher in grape farmers than in controls, after adjusting for age and sex. Sensitization to pollens (OR, 4.20; 95% CI, 1.41-12.82, P<0.01) and allergic rhinitis (OR, 3.06; 95% CI, 1.21-8.28, P<0.05) were found to be significantly associated with self-reported itchy rash in the grape farmers group. CONCLUSIONS: Grape farmers reported skin symptoms more frequently than non exposed controls, and IgE-mediated sensitization to pollens was found to be significantly associated with the reported symptoms. Further studies are needed to evaluate the impact of specific occupational agents on skin diseases among grape farmers. PMID- 16362953 TI - Regulation of microtubule dynamics in 3T3 fibroblasts by Rho family GTPases. AB - To get insight into the action of Rho GTPases on the microtubule system we investigated the effects of Cdc42, Rac1, and RhoA on the dynamics of microtubules in Swiss 3T3 fibroblasts. In control cells microtubule ends were dynamic: plus ends frequently switched between growth, shortening and pauses; the growth phase predominated over shortening. Free minus ends of microtubules depolymerized rapidly and never grew. Free microtubules were short-lived, and the microtubule network was organized into a radial array. In serum-starved cells microtubule ends became more stable: although plus ends still transited between growth and shortening, polymerization and depolymerization excursions became shorter and balanced each other. Microtubule minus ends were also stabilized. Consequently lifespan of free microtubules increased and microtubule array changed its radial pattern into a random one. Activation of Cdc42 and Rac1 in serum-starved cells promoted dynamic behavior of microtubule plus and minus ends, while inhibition of these GTPases in serum-grown cells suppressed microtubule dynamics and mimicked all effects of serum starvation. Activation of RhoA in serum-grown cells had effects similar to Cdc42 /Rac1 inactivation: it suppressed the dynamics of plus and minus ends, reduced the length of growth and shrinking episodes, and disrupted the radial organization of microtubules. However, in contrast to Cdc42 and Rac1 inactivation, active RhoA had no effect on the balance between microtubule growth and shortening. We conclude that Cdc42 and Rac1 have similar stimulating effects on microtubule dynamics while RhoA acts in an opposite way. PMID- 16362954 TI - Comparison of beta-tubulin mRNA and protein levels in 12 human cancer cell lines. AB - Antimitotic drugs are chemotherapeutic agents that bind tubulin and microtubules. Resistance to these drugs is a major clinical problem. One hypothesis is that the cellular composition of tubulin isotypes may predict the sensitivity of a tumor to antimitotics. Reliable and sensitive methods for measuring tubulin isotype levels in cells and tissues are needed to address this hypothesis. Quantitative measurements of tubulin isotypes have frequently relied upon inferring protein amounts from mRNA levels. To determine whether this approach is justified, protein and mRNA levels of beta-tubulin isotypes from 12 human cancer cell lines were measured. This work focused on only beta-tubulin isotypes because we had readily available monoclonal antibodies for quantitative immunoblots. The percentage of beta-tubulin isotype classes I, II, III, and IVa + IVb mRNA and protein were compared. For beta-tubulin class I that comprises >50% of the beta tubulin protein in 10 of the 12 cell lines, there was good agreement between mRNA and protein percentages. Agreement between mRNA and protein was also found for beta-tubulin class III. For beta-tubulin classes IVa + IVb, we observed higher protein levels compared to mRNA levels.Beta-tubulin class II protein was found in only four cell lines and in very low abundance. We conclude that quantitative Western blotting is a reliable method for measuring tubulin isotype levels in human cancer cell lines. Inferring protein amounts from mRNA levels should be done with caution, since the correspondence is not one-to-one for all tubulin isotypes. PMID- 16362955 TI - Ca2+-dependent in vitro contractility of a precipitate isolated from an extract of the heliozoon Actinophrys sol. AB - Contraction of axopodia in actinophrid heliozoons (protozoa) is induced by a unique contractile structure, the "contractile tubules structure (CTS)". We have previously shown that a cell homogenate of the heliozoon Actinophrys sol yields a precipitate on addition of Ca2+ that is mainly composed of filamentous structures morphologically identical to the CTS. In this study, to further characterize the nature of the CTS in vitro, biochemical and physiological properties of the precipitate were examined. SDS-PAGE analysis showed that the Ca2+-induced precipitate was composed of many proteins, and that no proteins in the precipitate showed any detectable changes in electrophoretic mobility on addition of Ca2+. Addition of extraneous proteins such as bovine serum albumin to the cell homogenate resulted in cosedimentation of the proteins with the Ca2+-induced precipitate, suggesting that the CTS has a high affinity for other proteins that are not related to precipitate formation. Appearance and disappearance of the precipitate were repeatedly induced by alternating addition of Ca2+ and EGTA, and its protein composition remained unchanged even after repeated cycles. When adhered to a glass surface, the precipitate showed Ca2+-dependent contractility with a threshold of 10-100 nM, and this contractility was not inhibited by colchicine or cytochalasin B. The precipitate repeatedly contracted and relaxed with successive addition and removal of Ca2+, indicating that the contraction was controlled by Ca2+ alone with no need for any other energy supply. From our characterization of the precipitate, we concluded that its Ca2+-dependent formation and contraction are associated with the unique contractile organelle, the "contractile tubules structure". PMID- 16362956 TI - First steps towards tissue engineering of small-diameter blood vessels: preparation of flat scaffolds of collagen and elastin by means of freeze drying. AB - Porous scaffolds composed of collagen or collagen and elastin were prepared by freeze drying at temperatures between -18 and -196 degrees C. All scaffolds had a porosity of 90-98% and a homogeneous distribution of pores. Freeze drying at -18 degrees C afforded collagen and collagen/elastin matrices with average pore sizes of 340 and 130 mum, respectively. After 20 successive cycles up to 10% of strain, collagen/elastin dense films had a total degree of strain recovery of 70% +/- 5%, which was higher than that of collagen films (42% +/- 6%). Crosslinking of collagen/elastin matrices either in water or ethanol/water (40% v/v) was carried out using a carbodiimide (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, EDC) in combination with a succinimide (N-hydroxysuccinimide, NHS) in the presence or absence of a diamine (J230) or by reaction with butanediol diglycidylether (BDGE), followed by EDC/NHS. Crosslinking with EDC/NHS or EDC/NHS/J230 resulted in matrices with increased stiffness as compared to noncrosslinked matrices, whereas sequential crosslinking with the diglycidylether and EDC/NHS yielded very brittle scaffolds. Ethanol/water was the preferred solvent in the crosslinking process because of its ability to preserve the open porous structure during crosslinking. Smooth muscle cells were seeded on the (crosslinked) scaffolds and could be expanded during 14 days of culturing. PMID- 16362957 TI - Sustained-release delivery systems of triclosan for treatment of Streptococcus mutans biofilm. AB - Dental diseases are chronic infections caused by oral bacteria harboring the dental biofilm. Local sustained-release delivery systems prolong the duration of a drug in the oral cavity, thus enhancing its therapeutic potential, while reducing its side effects. Triclosan is an agent that was found to have an antibacterial effect against oral bacteria. However, its substantivity in the oral cavity is low, resulting in reduced antibacterial efficiency. The purpose of this study was to develop a local sustained release device containing triclosan and to test its antibacterial efficacy on Streptococcus mutans biofilm. Our results show that we can formulate an ethylcellulose-based, nondegradable, sustained-release device in which 80% of the loaded triclosan is released over a 10-day period. The release rate of triclosan corresponded to the Higuchi's planar homogenous diffusion release model (r2 = 0.998). A degradable local sustained release delivery based on a methacrylate ester matrix was also developed for a faster release rate of triclosan. The release kinetics in those types of sustained-release delivery systems was erosion control. The local sustained release delivery system significantly affected the viability of S. mutans in biofilm compared to placebo as was tested by confocal laser scanning microscopy. Our in vitro results show that triclosan can be incorporated into degradable or nondegradable sustained-release drug delivery systems. The release of triclosan from the local sustained-release delivery system can be controlled, thus extending its antibacterial properties. PMID- 16362958 TI - Healing pattern of bone defects covered by different membrane types--a histologic study in the porcine mandible. AB - Few investigations on guided bone regeneration (GBR) focus on the behaviour of tissues adjacent to barrier membranes. This study was conducted to (1) evaluate the barrier function potential of different resorbable and nonresorbable membranes for GBR, (2) investigate their structural changes after different intervals, and (3) characterize tissue composition and reaction adjacent to the barrier by qualitative histologic evaluation. Seven barriers for GBR were used per animal (made of dense or expanded polytetrafluoroethylene (d/ePTFE), titanium, polyetherurethane, collagen and two polylactide-polyglycolide-/ trimethylenecarbonate-co-polymers (PLPG, LPGTC) in standardized defects not exceeding the critical size) without using bone substitution material or autogenous bone at the right inferior margin of the mandibles of six domestic pigs. Samples of the defect areas with membranes were harvested after 2 days (one animal), 4 and 8 (two animals, each) and 12 weeks (one animal), respectively. The healing of bone defects was completed in all animals after 12 weeks. Nonresorbable barriers prevented the soft tissue in-growth into standardized defects. Thinner layers of fibrous tissue were seen underneath the dense and rigid barriers (dPTFE, titanium) when compared with collagen and PLPG/LPGTC, in which soft-tissue plugs occupied the crestal defect portion. PLPG-/LPGTC-barriers underwent structural changes after 4 weeks and revealed blistered central layers, whereas structural changes were not evident in nonresorbable barriers. The degradation of PLPG-/LPGTC-membranes was present with in-growth of fibres, vessels, and cells. Using collagen or synthetic polymer barriers for GBR, the application of bone or bone substitutes to prevent membrane prolapse into the defect is suggested. PMID- 16362959 TI - Comparison of two radio-frequency plasma sterilization processes using microspot evaluation of microbial inactivation. AB - In this study, we evaluated gas plasma surface sterilization methods in a specific sterilizer. We have introduced a new monitoring method using 0.4 microm pore size membranes, which in this study gave the information corresponding to 3000 exposed biological indicators per treatment cycle. This enabled us to compare the fraction of inoculates that showed no growth after exposure for 30 different locations in the chamber, and hereby identify weak and strong spots in the chamber with regard to sporicidal effect. Membranes were also used to expose a broad spectrum of soil bacteria for plasma treatment at four different conditions. The organisms were identified using PCR and sequencing. The test showed that Bacillus stearothermophilus spores were inactivated at the slowest rate among the tested microorganisms. Further alpha-proteobacteria (Gram negative) seemed more sensitive than the rest of the tested organisms. The microspot evaluation approach has been a most useful tool in the assessment of sterilization performance in sterilizers that do not have clear measurable parameters related to the sterilization. PMID- 16362960 TI - In vivo performance of two different hydroxyapatite coatings on titanium prepared by discharging in electrolytes. AB - This study reports a discharging method for bone-like carbonated HA (cHA)-coating (Ca/P 1.71) and stoichiometric HA (sHA)-coating (Ca/P 1.67) with micrometer order thicknesses on titanium plates, using modified body fluid and acidic calcium phosphate solutions, respectively. In vivo histological performance of the HA coatings prepared by discharging in electrolytes was evaluated. Bone-contact indexes of HA coatings were measured microscopically. Additionally, bone-coating interface was analyzed by scanning electron microscopy and the use of an electron probe microanalyzer. Results demonstrated that there was no significant difference in contact index between HA coatings. However, the cHA coating was practically replaced by immature bone, and the titanium metal substrate was directly connected to the bone structure whereas the sHA coating layer remained and was partially detached from the titanium metal substrate. Since detached coating particles are pathogens, and can cause peri-implantitis, the cHA coating was more favorable than the sHA coating even if contact index was equivalent to that of the sHA coating. It is thought that coating thickness and chemical composition of coatings are important for biological stability of implants. In conclusion, since bone-like thin cHA coating showed high osteoconductivity and bone replacement, bone-like HA is superior to sHA coating for use in dental implants. PMID- 16362961 TI - Preliminary beta-tricalcium phosphate coating prepared by discharging in a modified body fluid enhances collagen immobilization onto titanium. AB - Because of its excellent scaffold properties toward bone cells, collagen has been recognized as a promising extracellular matrix protein for surface modification of titanium implants. Hydroxyapatite (HA) coatings have been investigated as a preliminary coating for collagen immobilization on titanium implants. However, the composition of HA-collagen is recognized as being difficult, and while many studies have suggested that biodegradable beta-tricalcium phosphate (beta-TCP) has better osteoconductivity than HA, the efficiency of preliminary beta-TCP coating for collagen immobilization on titanium surfaces has yet to be evaluated. This investigation aimed to evaluate the applicability of HA and beta-TCP coatings, prepared by discharging in modified body fluids, as preliminary collagen coatings. To increase collagen induction on preliminary HA and beta-TCP coatings, we used a new cathodic polarization method. X-ray photoelectron spectroscopy revealed that the bonding strength between the collagen NH(+) amino groups of collagen and phosphate (PO(4) (3-)) was greater on the beta-TCP coating than the HA coating. The preliminary beta-TCP coating was tightly crosslinked with RCOO(-) carboxyl groups of the collagen molecules and showed high cellular responses, even in the early stage of cell cultivation. Thus, this coating was found to be more effective than HA as a preliminary coating for collagen immobilization on titanium implants. PMID- 16362962 TI - Effects of additives on the rheological properties and injectability of a calcium phosphate bone substitute material. AB - An injectable calcium phosphate bone substitute material has been prepared by mixing amorphous calcium phosphate (ACP) and dicalcium phosphate dihydrate (DCPD) for use in noninvasive surgery, and the influence of additives, such as disodium hydrogen phosphate, polyethylene glycol (PEG), glycerin, and citric acid, on the rheological properties and injectability of the ACP + DCPD cement system have been studied in this work. Novel approach of thixotropy measurement has been used to characterize the stability of the pastes. The results show that the injectability and the setting time can be augmented by the addition of disodium phosphate solution to the paste but reduced by the addition of PEG 200, glycerin, or citric acid to the paste. This study suggests that the injectability and the setting time of the ACP + DCPD bone substitute material can be balanced, and the injectable calcium phosphate bone substitute material with satisfied fluidity and injectability for clinical operation can be prepared by optimizing the additives and their concentrations, according to different clinical requirements. PMID- 16362963 TI - An improved hydrophilicity via electrospinning for enhanced cell attachment and proliferation. AB - The wettability of electrospun poly(epsilon-caprolactone) (PCL) mats was improved by co-electrospinning with poly(vinyl alcohol) (PVA), by double-spinneret electrospinning method. The improved hydrophilicity of the hybrid PCL/PVA mats was confirmed by water contact angle measurement. The in vitro cell attachment on the hydrophobic PCL and hydrophilically modified PCL/PVA mats was compared by culture studies using human prostate epithelial cells (HPECs). The stability of water-soluble PVA component in the electrospun PCL/PVA mats was checked by thermogravimetric analysis and intensity of fluorescence material after immersion in water for 7 days. The images from scanning electron microscopy, field emission scanning electron microscopy, and optical microscopy showed that the attachment and proliferation rate of HPECs were improved by introducing PVA into the electrospun PCL mats. PMID- 16362964 TI - Hydrogen absorption of titanium and nickel-titanium alloys during long-term immersion in neutral fluoride solution. AB - Hydrogen absorption of biomedical titanium and Ni-Ti alloys in a neutral fluoride (2.0% NaF) solution for up to 10,000 h at 37 degrees C has been evaluated by means of hydrogen thermal desorption analysis. For alpha titanium (commercial pure titanium), the amount of absorbed hydrogen was, at most, 10-30 mass ppm, and the corrosion product and hydride formation were revealed on the surface of the specimen by X-ray diffraction analysis. Ni-Ti superelastic alloy absorbed approximately 150 mass ppm of hydrogen, which was probably sufficient to result in the pronounced degradation of the mechanical properties, although corrosion was hardly observed. In contrast, hydrogen absorption of alpha-beta titanium (Ti 6Al-4V) and beta titanium (Ti-11.3Mo-6.6Zr-4.3Sn) alloys was negligible, although general corrosion was observed. The results of the present study indicate that the susceptibility of titanium and Ni-Ti alloys to hydrogen absorption in the neutral fluoride solution is different from that in the acidic fluoride solution reported previously. PMID- 16362965 TI - Adsorption of biometals to monosodium titanate in biological environments. AB - Monosodium titanate (MST) is an inorganic sorbent/ion exchanger developed for the removal of radionuclides from nuclear wastes. We investigated the ability of MST to bind Cd(II), Hg(II), Au(III), or the Au-organic compound auranofin to establish the utility of MST for applications in environmental decontamination or medical therapy (drug delivery). Adsorption isotherms for MST were determined at pH 7-7.5 in water or phosphate-buffered saline. The extent of metal binding was determined spectroscopically by measuring the concentrations of the metals in solution before and after contact with the MST. Cytotoxic responses to MST were assessed using THP1 monocytes and succinate dehydrogenase activity. Monocytic activation by MST was assessed by TNFalpha secretion (ELISA) with or without lipopolysaccharide (LPS) activation. MST adsorbed Cd(II), Hg(II), and Au(III) under conditions similar to those in physiological systems. MST exhibited the highest affinity for Cd(II) followed by Hg(II) and Au (III). MST (up to 100 mg/L) exhibited only minor (<25% suppression of succinate dehydrogenase) cytotoxicity and did not trigger TNFalpha secretion nor modulate LPS-induced TNFalpha secretion from monocytes. MST exhibits high affinity for biometals with no significant biological liabilities in these introductory studies. MST deserves further scrutiny as a substance with the capacity to decontaminate biological environments or deliver metals or metal compounds for therapeutic applications. PMID- 16362966 TI - Characterization of a faster resorbing polymer after real time aging. AB - This study evaluated the in vitro strength retention and polymer characteristics of specimens made from commercially available 85:15 poly(D,L-lactide-co glycolide). Test samples included dogbone tensile specimens with a nominal thickness of either 0.75 and 1.0 mm, which were machined from compression-molded sheets, and screws with a major diameter of 2.71 mm and minor diameter of 2.14 mm, which were manufactured by injection molding. All samples were sterilized by e-beam irradiation prior to in vitro aging following a standard methodology. Mechanical testing and polymer analysis were performed at time zero and weekly up to 15 weeks of real time aging. The time zero maximum tensile strength of the 0.75 mm dogbone specimens averaged 55.86 +/- 0.72 MPa. The 1.0-mm dogbone specimens tested at time zero had an average maximum tensile strength of 34.55 +/ 0.36 MPa. The 0.75-mm and 1.0-mm thick dogbone specimens exhibited a controlled decrease in their tensile strength. The initial shear strength of the injection molded screws was 32.86 +/- 4.15 MPa. After 3 weeks of real time in vitro aging, the screws maintained approximately 70% of their initial (time zero) strength. The inherent viscosity and molecular weight (Mw) at time zero averaged approximately 0.9 dL/g and 98,000 g/mol respectively, and decreased at similar rates for both dogbones and screws. These results demonstrate a controlled, rapid degradation in the mechanical properties of 85:15 poly(D,L-lactide-co-glycolide) material, with sufficient strength for pediatric craniofacial applications. PMID- 16362967 TI - Effects of applied voltages on hydroxyapatite coating of titanium by electrophoretic deposition. AB - Hydroxyapatite (HA) coatings were deposited on titanium substrates by electrophoretic deposition (EPD) at constant voltage and dynamic voltage, respectively. Various surface morphologies were observed under different type of voltages. Under a constant voltage of 20 V, a dense HA coating could be prepared. Under a constant voltage of 200 V, big HA particles were deposited and the coating was porous. Under a dynamic voltage, a continuous gradient HA coating could be obtained. HA coatings were characterized with a field emission-scanning electron microscopy (FE-SEM) and an X-ray diffraction (XRD). XRD indicated no significant HA decomposition when the coatings were sintered for 2 h at 800 degrees C. PMID- 16362968 TI - Timing of entry of meiosis depends on a mark generated by DNA methyltransferase 3a in testis. AB - Reprogramming of DNA methylation is an essential part of gametogenesis, and a role of two members of the DNA methyltransferase (Dnmt) family, Dnmt3a and Dnmt3L, has been recognized. In an attempt to elucidate the role of Dnmt3a, we analyzed the progression of spermatogenesis in Dnmt3a (-/-) homozygotes during the first 3 weeks of post-natal development. The emerging picture was markedly different from that recently reported for the Dnmt3L protein. In the Dnmt3a (-/-) testis, at the expected time of entry into meiosis (11-13 dpp), the number of spermatocytes was greatly reduced. They progressively accumulated during the following days, but at a slower rate than in the wild type. Once started, however, the pachytene stage was apparently completed with normal chromosome pairing and formation of the sex vesicle, and spermatogenesis further progressed with the appearance and the expression of round spermatid specific markers. Interestingly and unlike Dnmt3L (-/-) spermatocytes, Dnmt3a (-/-) germ cells showed only a minor reduction in the methylation of interspersed repetitive elements and retroposons. The Dnmt3a might thus generate a mark important for the initiation of male meiosis that is distinct from that created by Dnmt3L. PMID- 16362969 TI - Quantitative expression of candidate genes for developmental competence in bovine two-cell embryos. AB - Only competent oocytes are able to undergo complete maturation and normal embryonic development. Therefore, the identification of genes that are differentially expressed in competent oocytes would contribute to our understanding of the factors controlling competency. It is well known that time of cleavage after insemination in vitro is highly correlated with embryonic developmental potential and this can be used to distinguish between oocytes of different quality. The main objective of this study was to identify genes associated with competency and rapid cleavage. We examined the expression of 16 candidate genes (IDH, YEAF Cathepsin B, RAD50, TCP1 NCOR1, HUEL, STK6, ZNF403, AOP2, EEF1A1, Hsp90, Hsp40, AKR1B1, PGRMC1, and DMRT2) in early and late cleaving embryos, by real time PCR. These transcripts were derived from previous study in our laboratory using cDNA coming from a suppressive subtraction hybridization (SSH) between early cleaving versus late cleaving embryos spotted on a microarray slide. Of the 16 genes evaluated, 3 (IDH, YEAF, and H2A) showed statistical difference (P < 0.05) between early and late cleaving embryos. However, some genes such as Cathepsin B (P = 0.0677), RAD50 (P = 0.0899), and TCP1 (P = 0.0824) tended to show higher expression in the early cleaving than in the late cleaving embryo. In conclusion, we have identified three genes (YEAF, IDH, H2A) that were differentially expressed in the early cleaving embryos, and their expression can be associated with greater developmental competence. PMID- 16362970 TI - Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos. AB - DNA methylation/demethylation of donor genomes in recipient ooplasm after nuclear transfer occurs in a species-specific way. In cloned rabbit and bovine embryos, repetitive sequences maintain the donor-type methylation status, but typical demethylation of repetitive sequences takes place in cloned porcine embryos. To clarify whether the demethylation is controlled by donor nucleus intrinsic property or by recipient ooplasm, we used interspecies somatic cell nuclear transfer (iSCNT) model to examine the methylation status of repetitive sequences in pig-to-rabbit and rabbit-to-pig interspecies embryos. We found that no demethylation of pig repetitive sequences was observed in pig-to-rabbit iSCNT embryos, while the examined rabbit repetitive sequence Rsat IIE was demethylated in rabbit-to-pig iSCNT embryos. These results indicate that demethylation of donor repetitive sequences is determined by ooplasm but not by donor intrinsic property and that ooplasm from different species have different capabilities to demethylate genes. PMID- 16362971 TI - Spetex-1: a new component in the middle piece of flagellum in rodent spermatozoa. AB - Spetex-1 has recently been isolated by differential display and screening of cDNA library. It encodes a protein of 556 amino acid residues possessing coiled-coil motifs. In the rat seminiferous tubules (ST), Spetex-1 was expressed in the cytoplasm of elongating spermatids. To examine the subcellular distribution of Spetex-1 in mature spermatozoa, we performed biochemical and immunocytochemical approaches. We found that Spetex-1 that was synthesized in the cytoplasm of elongating spermatids was subsequently integrated as a middle piece component into spermatozoa during spermiogenesis. After integration, the majority of Spetex 1 in spermatozoa could be extracted by 6M urea under reduced condition but not released by the treatment of 1% Triton X-100. Immunoelectron microscopy demonstrated that Spetex-1 seemed to locate at the inner side of outer dense fibers (ODFs) in the middle piece or the narrow space between ODFs and axoneme. Spetex-1 might be involved in the stability of the structural complexity comprising axoneme and ODFs in the middle piece of sperm flagellum. PMID- 16362972 TI - Suprazero cooling conditions significantly influence subzero permeability parameters of mammalian ovarian tissue. AB - To model the cryobiological responses of cells and tissues, permeability characteristics are often measured at suprazero temperatures and the measured values are used to predict the responses at subzero temperatures. The purpose of the present study was to determine whether the rate of cooling from +25 to +4 degrees C influenced the measured water transport response of ovarian tissue at subzero temperatures in the presence or absence of cryoprotective agents (CPAs). Sections of freshly collected equine ovarian tissue were first cooled either at 40 degrees C/min or at 0.5 degrees C/min from 25 to 4 degrees C, and then cooled to subzero temperatures. A shape-independent differential scanning calorimeter (DSC) technique was used to measure the volumetric shrinkage during freezing of equine ovarian tissue sections. After ice was induced to form in the extracellular fluid within the specimen, the sample was frozen from the phase change temperature to -50 degrees C at 5 degrees C/min. Replicate samples were frozen in isotonic medium alone or in medium containing 0.85 M glycerol or 0.85 M dimethylsulfoxide. The water transport response of ovarian tissue samples cooled at 40 degrees C/min from 25 to 4 degrees C was significantly different (confidence level >95%) from that of tissue samples cooled at 0.5 degrees C/min, whether in the presence or absence of CPAs. We fitted a model of water transport to the experimentally-derived volumetric shrinkage data and determined the best fit membrane permeability parameters (L(pg) and E(Lp)) of equine ovarian tissue during freezing. Subzero water transport parameters of ovarian tissue samples cooled at 0.5 degrees C/min from 25 to 4 degrees C ranged from: L(pg) = 0.06 to 0.73 microm/min.atm and E(Lp) = 6.1 to 20.5 kcal/mol. The corresponding parameters of samples cooled at 40 degrees C/min from 25 to 4 degrees C ranged from: L(pg) = 0.04 to 0.61 microm/min.atm and E(Lp) = 8.2 to 54.2 kcal/mol. Calculations made of the theoretical response of tissue at subzero temperatures suggest that the optimal cooling rates to cryopreserve ovarian tissue are significantly dependent upon suprazero cooling conditions. PMID- 16362973 TI - Differential response of cumulus cell-enclosed and denuded mouse oocytes in a meiotic induction model system. AB - In this study we have examined the effects of denuded oocyte coculture with dissociated cumulus cells (CC) or intact oocyte-CC complexes on meiotic resumption. When denuded oocytes (DO) or cumulus cell-enclosed oocytes (CEO) were cultured in 40-microl drops of medium under oil, and held in meiotic arrest with 4 mM hypoxanthine plus 25 microM dbcAMP, they underwent germinal vesicle breakdown (GVB) at similar frequencies (34%-35%). Coculture of DO with complexes or dissociated CCs stimulated maturation (50% and 61% GVB, respectively), with no effect of DO on maturation of cocultured CEO (32% GVB). This coculture effect was increased with the number of CCs added to the culture drop. When either glucose or glutamine was eliminated from the medium, no meiotic induction resulted from cocultured CCs. When CEO were cultured alone in microdrops, increasing their number from 10 to 50 significantly lowered the percentage resuming maturation, an effect also reduced by removing glucose and/or glutamine from the medium. This effect was not observed with DO. When inhibitory medium was conditioned overnight with complexes, subsequent culture with DO led to higher maturation percentages than culture in unconditioned medium; however, when CEO were cultured in conditioned medium, there was either no effect or increased inhibition of maturation. Assay of glucose and pyruvate in spent medium showed that DO cultured alone consumed glucose and pyruvate, but under CC coculture conditions more glucose was consumed and significant amounts of pyruvate accumulated in the medium, changes that led to an increase in the maturation of DO. Further experiments showed that DO were more sensitive than CEO to the meiosis-inducing effect of pyruvate. These results demonstrate different responsiveness of DO and CEO to coculture conditions and question the physiological relevance of denuded oocyte/CC coculture to study meiotic induction. PMID- 16362974 TI - Utilization of citrate and lactate through a lactate dehydrogenase and ATP regulated pathway in boar spermatozoa. AB - Incubation of boar spermatozoa in Krebs-Ringer-Henseleit medium with either 10 mM lactate or 10 mM citrate induced a fast and robust increase in the intracellular levels of ATP in both cases, which reached a peak after 30 sec of incubation. Utilization of both citrate and lactate resulted in the export of CO(2) to the extracellular medium, indicating that both substrates were metabolized through the Krebs cycle. Incubation with citrate resulted in the generation of extracellular lactate, which was inhibited in the presence of phenylacetic acid. This indicates that lactate is produced through the pyruvate carboxylase step. In addition, there was also a significant increase in tyrosine phosphorylation induced by both citrate and lactate. Boar sperm has a sperm-specific isoform of lactate dehydrogenase (LDH), mainly located in the principal piece of the tail. Kinetic studies showed that boar sperm has at least two distinct LDH activities. The major activity (with an estimated Km of 0.51 mM) was located in the supernatants of sperm extracts. The minor LDH activity (with an estimated Km of 5.9 mM) was associated with the nonsoluble fraction of sperm extracts. Our results indicate that boar sperm efficiently metabolizes citrate and lactate through a metabolic pathway regulated by LDH. PMID- 16362975 TI - Noninvasive ventilation during gastrostomy tube placement in patients with severe duchenne muscular dystrophy: case reports and review of the literature. AB - Individuals with Duchenne muscular dystrophy may benefit from gastrostomy tube feeding due to progressive dysphagia and malnutrition. However, due to their severely impaired pulmonary function, these individuals are at risk of severe complications when they are sedated or undergo anesthesia for the procedure. We previously described a technique of noninvasive positive pressure ventilation to provide respiratory support during gastrostomy tube placement in such patients, but this technique had risks and limitations. In this case report, we examine two alternative techniques we used to provide respiratory support successfully to patients with severe muscular dystrophy and malnutrition who underwent percutaneous endoscopic gastrostomy tube placement. We then review the literature and discuss the potential benefits, risks, and limitations of the above techniques and of other options for gastrostomy placement in people with severe muscular dystrophy. PMID- 16362976 TI - Cytokeratin 20 expression identifies a subtype of pancreatic adenocarcinoma with decreased overall survival. AB - BACKGROUND: Cytokeratins are markers of epithelial cell differentiation useful in determining histogenesis for malignancies with an unknown primary. Application of this principle to a single malignancy may identify cancer subtypes with altered developmental programs. Herein, we investigate the relevance of two widely used cytokeratins (CKs), 7 and 20, to subtype pancreas cancer and identify associations with clinical features. METHODS: A tissue microarray was constructed using tumor specimens from 103 patients who underwent resection for pancreatic adenocarcinoma with curative intent. A subset of resection specimens was evaluated for pancreatic intraepithelial neoplasia (PanIN) lesions. Tissues were immunostained by using specific anticytokeratin 7 and 20 monoclonal antibodies. RESULTS: CK 7 and 20 expression was present in 96% and 63% cases of pancreatic adenocarcinoma, respectively. Ubiquitous CK 7 expression precluded further analysis. Tumoral CK 20 expression was not associated with any histopathologic parameter but correlated with worse prognosis when considered as either a dichotomous (P=0.0098) or continuous (P=0.007) variable. In a multivariate model, tumoral CK 20 expression remained a significant independent prognosticator. CK 20 expression was absent in all PanIN lesions from eight resection specimens in which the tumor component was negative for CK 20. In contrast, presence of tumoral CK 20 was highly concordant with its expression in corresponding PanINs. CONCLUSIONS: CK 20 expression defines a subtype of pancreas cancer with important biologic properties. When present, CK 20 expression is an early event in pancreatic carcinogenesis identifiable in precursor lesions. Further studies to identify the underlying genetic changes associated with this altered developmental pathway are warranted. PMID- 16362977 TI - Subsequent therapy can be administered after tositumomab and iodine I-131 tositumomab for non-Hodgkin lymphoma. AB - BACKGROUND: Iodine I-131 tositumomab is a well tolerated and effective therapy for recurrent low-grade and transformed low-grade non-Hodgkin lymphoma (NHL). Hematologic reserve after radioimmunotherapy (RIT) is an important consideration when subsequent therapy is required. METHODS: One hundred fifty-five patients who received treatment with I-131 tositumomab were assessed, and 68 patients had progressive disease after RIT. The median age (n=68 patients) was 59 years (range,18-82 yrs), and patients received a median of 2 pre-RIT regimens (range,1 8 regimens), including 66% who received anthracycline, 19% who received platinum, and 50% who received fludarabine. RESULTS: The median time to disease progression (among progressors) was 168 days (range, 19-771 days). At the time they developed recurrent disease, patients had median white blood cell count (WBC) of 4.9 K cells/microL (range, 1.1-21.4 K cells/microL), a median absolute neutrophil count (ANC) of 3.25 K cells/microL (range, 0.59-8.20 K cells/microL), a median platelet count of 130 K cells/microL (range, 9-440 K cells/microL), and there was no significant difference between pre-RIT and recurrence values except for the platelet count (P<0.05). No patient demonstrated a WBC<1.0 K cells/microL or an ANC<0.5 K cells/microL, although 1 patient had a platelet count<10 K cells/microL. Twenty-four patients subsequently received no further chemotherapy; and, in 21 patients (88%), hematologic parameters appeared to allow subsequent chemotherapy if necessary (blood counts: National Cancer Institute Grade 0-2). Among 44 patients (65%) who received further chemotherapy (median, 2 regimens; range, 1-4 regimens), 19 patients (43%) were treated with anthracyclines, 17 patients (39%) were treated with platinum, 10 patients (23%) were treated with fludarabine, and 13 patients (30%) underwent stem cell transplantation. Disease improvement occurred in most patients, although 18 patients died (40%) after further chemotherapy, predominantly from refractory lymphoma. CONCLUSIONS: Most patients with progressive disease after treatment with iodine I-131 tositumomab were able to receive subsequent therapy, including cytotoxic chemotherapy and stem cell transplantation. PMID- 16362978 TI - CpG island hypermethylation in progression of esophageal and gastric cancer. AB - The upper gastrointestinal (GI) cancers have various carcinogenic pathways and precursor lesions, such as dysplasia for esophageal squamous cell carcinoma, Barrett esophagus for esophageal adenocarcinoma, and intestinal metaplasia for the intestinal-type of gastric cancer. Recently, many epigenetic events in carcinogenic pathways have been revealed, along with genomic and genetic alterations. This information has provided deeper insight into an understanding of the mechanisms of upper GI carcinogenesis. Moreover, detection methods of aberrant methylation have been applied to clinical fields to stratify high-risk groups, detect early cancer, and to predict clinical outcomes. In this review, a variety of information is summarized regarding gene hypermethylation in esophageal and gastric cancer. PMID- 16362979 TI - The role of backbone stability near Ala44 in the high reduction potential class of rubredoxins. AB - Rubredoxins may be separated into high and low reduction potential classes, with reduction potentials differing by approximately 50 mV. Our previous work showed that a local shift in the polar backbone due to an A(44) versus V(44) side-chain size causes this reduction potential difference. However, this work also indicated that in the low potential Clostridium pasteurianum (Cp) rubredoxin, a V(44) --> A(44) mutation causes larger local backbone flexibility, because the V(44) side-chain present in the wild-type (wt) is no longer present to interlock with neighboring residues to stabilize the subsequent G(45). Since Pyrococcus furiosus (Pf) and other high potential rubredoxins generally have a P(45), it was presumed that a G(45) --> P(45) mutation might stabilize a V(44) --> A(44) mutation in Cp rubredoxin. Here crystal structure analysis, energy minimization, and molecular dynamics (MD) were performed for wt V(44)G(45), single mutant A(44)G(45) and double mutant A(44)P(45) Cp, and for wt A(44)P(45) Pf rubredoxins. The local structural, dynamical, and electrostatic properties of Cp gradually approach wt Pf in the order wt Cp to single to double mutant because of greater sequence similarity, as expected. The double mutant A(44)P(45) Cp exhibits increased backbone stability near residue 44 and thus enhances the probability that the backbone dipoles point toward the redox site, which favors an increase in the electrostatic contribution to the reduction potential. It appears that the electrostatic potential of residue 44 and the solvent accessibility to the redox are both determinants for the reduction potentials of homologous rubredoxins. Overall, these results indicate that an A(44) in a rubredoxin may require a P(45) for backbone stability whereas a V(44) can accommodate a G(45), since the valine side-chain can interlock with its neighbors. PMID- 16362980 TI - The pleiotropic effects of Kaposi's sarcoma herpesvirus. AB - Kaposi's sarcoma herpesvirus (KSHV), or human herpesvirus 8 (HHV8), is an essential factor in the pathogenesis of Kaposi's sarcoma (KS), multicentric Castleman's disease (MCD), and primary effusion lymphoma (PEL). Case reports suggest an occasional involvement in bone marrow hypoplasia and haemophagocytic syndrome, but other disease associations are unconfirmed or controversial. KSHV associated disease is of particular importance in immunosuppressed individuals, in particular in patients with HIV infection and transplant recipients. KSHV establishes a latent infection in the majority of infected cells in KS, MCD, and PEL, but lytic replication occurs in a small fraction of infected cells. Viral proteins expressed during both the latent and the lytic phase of the viral life cycle contribute to the pathogenesis of KSHV-associated diseases. PMID- 16362981 TI - Morbilliviruses and human disease. AB - Morbilliviruses are a group of viruses that belong to the family Paramyxoviridae. The most instantly recognizable member is measles virus (MV) and individuals acutely infected with the virus exhibit a wide range of clinical symptoms ranging from a characteristic mild self-limiting infection to death. Canine distemper virus (CDV) and rinderpest virus (RPV) cause a similar but distinctive pathology in dogs and cattle, respectively, and these, alongside experimental MV infection of primates, have been useful models for MV pathogenesis. Traditionally, viruses were identified because a distinctive disease was observed in man or animals; an infectious agent was subsequently isolated, cultured, and this could be used to recapitulate the disease in an experimentally infected host. Thus, satisfying Koch's postulates has been the norm. More recently, particularly due to the advent of exceedingly sensitive molecular biological assays, many researchers have looked for infectious agents in disease conditions for which a viral aetiology has not been previously established. For these cases, the modified Koch's postulates of Bradford Hill have been developed as criteria to link a virus to a specific disease. Only in a few cases have these conditions been fulfilled. Therefore, many viruses have over the years been definitely and tentatively linked to human diseases and in this respect the morbilliviruses are no different. In this review, human diseases associated with morbillivirus infection have been grouped into three broad categories: (1) those which are definitely caused by the infection; (2) those which may be exacerbated or facilitated by an infection; and (3) those which currently have limited, weak, unsubstantiated or no credible scientific evidence to support any link to a morbillivirus. Thus, an attempt has been made to clarify the published data and separate human diseases actually linked to morbilliviruses from those that are merely anecdotally associated. PMID- 16362982 TI - New insights into the function of granulomas in human tuberculosis. AB - The human tuberculous granuloma provides the morphological framework for local immune processes central to the outcome of tuberculosis. This review article describes investigations on human lung granulomas aimed at better understanding the regional host response and counter-measures to Mycobacterium tuberculosis. These findings lead to a revised view of the regional immune response in human tuberculosis. Novel insights into this dynamic cross-talk form the basis of novel intervention strategies. PMID- 16362983 TI - Pathogenesis and prevalence of variant Creutzfeldt-Jakob disease. AB - In the late 1980s and early 1990s, there was widespread exposure of the UK population to bovine spongiform encephalopathy (BSE)-contaminated food products, which has led to over 150 deaths from variant Creutzfeldt-Jakob disease (vCJD). Although the pathogenesis in humans is not fully understood, data from animal models and, to a lesser extent, patients with vCJD suggest that oral exposure to BSE is rapidly followed by accumulation of PrP(res) in gut-associated lymphoid tissue, then, after haematogenous spread, throughout the lymphoreticular system. Spread to the central nervous system may not occur for several years, but blood from individuals in the pre-clinical phase appears to be able to transmit disease. The incidence of vCJD has remained low and is in decline, but it is known from iatrogenic CJD and kuru that human prion disease can have incubation periods of up to 40 years. Cases of vCJD are therefore likely to occur for many more years and alternative phenotypes may develop in individuals with different PRNP genotypes to those seen to date. Studies in transgenic mice have shown that sub-clinical infection is frequent following oral exposure to BSE and a study looking at the accumulation of PrP in anonymized human lymphoid tissue samples found positive cases. There are likely to be a number of asymptomatic 'carriers' of disease within the UK and although it is unclear whether these individuals will develop clinical disease, there is a potential for iatrogenic spread to others. These uncertainties highlight the importance of developing a reliable blood test for vCJD and the continued need for surveillance. PMID- 16362984 TI - Subversion of cellular functions by Listeria monocytogenes. AB - Listeria monocytogenes is a Gram-positive bacterium that is able to invade and multiply within eukaryotic cells. Its intracellular life-cycle includes pathogen induced phagocytosis, lysis of the phagocytic vacuole, movement in the cytoplasmic environment, and a cell-to-cell spread mechanism. Many L. monocytogenes virulence factors have been studied in detail, certain of which subvert specific eukaryotic cell functions in order to favour infection. During entry, the invasion protein InlA takes advantage of the adhesion molecule E cadherin and the adherens junction machinery to adhere to target and invade polarized epithelial cells. Another invasion protein of the internalin family, InlB, subverts the signalling pathway of the hepatocyte growth factor receptor Met to induce endocytosis of the receptor and also to favour internalization of the bacteria in non-polarized epithelial cells. Once inside the cell, the haemolysin of L. monocytogenes--the listeriolysin O or LLO--is secreted to lyse the phagocytic vacuole, and when the bacteria is freed in the cytoplasm, the activity of the LLO is in part regulated by the infected cell itself, taking advantage of the pH sensitivity of the LLO that leads to its inactivation in the neutral eukaryotic cell cytoplasm. Finally, to induce bacterial movement in the cytoplasm, the L. monocytogenes surface protein ActA mimics the activity of the eukaryotic WASP family of proteins to recruit to the bacteria the actin nucleation machinery required for actin polymerization and for the formation of the actin structures (called 'actin comet tails') that propel the parasite in the cytosol and help it to invade neighbouring cells. PMID- 16362985 TI - The function of immunoglobulin A in immunity. AB - The vast surfaces of the gastrointestinal, respiratory, and genitourinary tracts represent major sites of potential attack by invading micro-organisms. Immunoglobulin A (IgA), as the principal antibody class in the secretions that bathe these mucosal surfaces, acts as an important first line of defence. IgA, also an important serum immunoglobulin, mediates a variety of protective functions through interaction with specific receptors and immune mediators. The importance of such protection is underlined by the fact that certain pathogens have evolved mechanisms to compromise IgA-mediated defence, providing an opportunity for more effective invasion. IgA function may also be perturbed in certain disease states, some of which are characterized by deposition of IgA in specific tissues. This review details current understanding of the roles played by IgA in both health and disease. PMID- 16362986 TI - Rational design of gene-based vaccines. AB - Vaccine development has traditionally been an empirical discipline. Classical vaccine strategies include the development of attenuated organisms, whole killed organisms, and protein subunits, followed by empirical optimization and iterative improvements. While these strategies have been remarkably successful for a wide variety of viruses and bacteria, these approaches have proven more limited for pathogens that require cellular immune responses for their control. In this review, current strategies to develop and optimize gene-based vaccines are described, with an emphasis on novel approaches to improve plasmid DNA vaccines and recombinant adenovirus vector-based vaccines. PMID- 16362987 TI - Bacterial gene therapy strategies. AB - The ability of bacteria to mediate gene transfer has only recently been established and these observations have led to the utilization of various bacterial strains in gene therapy. The types of bacteria used include attenuated strains of Salmonella, Shigella, Listeria, and Yersinia, as well as non pathogenic Escherichia coli. For some of these vectors, the mechanism of DNA transfer from the bacteria to the mammalian cell is not yet fully understood but their potential to deliver therapeutic molecules has been demonstrated in vitro and in vivo in experimental models. Therapeutic benefits have been observed in vaccination against infectious diseases, immunotherapy against cancer, and topical delivery of immunomodulatory cytokines in inflammatory bowel disease. In the case of attenuated Salmonella, used as a tumour-targeting vector, clinical trials in humans have demonstrated the proof of principle but they have also highlighted the need for the generation of strains with reduced toxicities and improved colonization properties. Altogether, the encouraging results obtained in the studies presented in this review justify further development of bacteria as a therapeutic vector against many types of pathology. PMID- 16362988 TI - The pathogenesis of syphilis: the Great Mimicker, revisited. AB - Syphilis is a chronic sexually transmitted infection caused by Treponema pallidum subspecies pallidum. Its protean clinical presentations earned it the name of the 'Great Mimicker'. Understanding of disease pathogenesis and how host-pathogen interactions influence the course of disease have been compromised by the facts that the organism cannot be grown in vitro and, as an exclusively human pathogen, inferences made from animal models are of limited applicability. Many questions remain about how T. pallidum biology contributes to distinctive features of syphilis, such as its ability to persist in the presence of a brisk host response or its propensity for neuro-invasion and congenital transmission. In 1998, the genome of T. pallidum was sequenced. The organism has a relatively small genome, suggesting that it utilizes host biosynthesis to fulfil some of its metabolic needs. While biological functions are suggested for only about 55% of T. pallidum's 1041 open reading frames, even these relatively early studies offer important insights into syphilis pathogenesis. A family of repeat genes, the Tp genes, encode proteins homologous to the major sheath proteins of T. denticola. Antibodies to the TprK variable regions are protective in a rabbit model. With successive passage, increasing diversity is observed in the TprK V region genes. Antigenic variation through gene conversion has been hypothesized to be one mechanism of escaping immune surveillance, allowing for prolonged infection and persistence in the presence of a robust host response. Human and animal studies suggest that a Th1 response is elicited in primary syphilis. Progression to the secondary stage is accompanied by a shift to a Th2 response, allowing for incomplete clearance of the pathogen. In pregnancy, intense inflammatory responses and prostaglandins induced by fetal infection may be responsible for fetal death or pre-term delivery and severe growth retardation or other manifestations of congenital syphilis. Understanding of the molecular targets of these immune responses may facilitate the development of vaccines for syphilis. PMID- 16362989 TI - Helicobacter infection and gastric neoplasia. AB - Chronic gastritis induced by Helicobacter pylori is the strongest known risk factor for adenocarcinoma of the distal stomach, yet only a minority of people who harbour this organism ever develop cancer. H. pylori isolates possess substantial genotypic diversity, which engenders differential host inflammatory responses that influence clinical outcome. H. pylori strains that possess the cag pathogenicity island and secrete a functional cytotoxin induce more severe gastric injury and further augment the risk for developing distal gastric cancer. However, carcinogenesis is also influenced by host genetic diversity, particularly involving immune response genes such as IL-1ss and TNF-alpha. It is important to gain insight into the pathogenesis of H. pylori-induced gastritis and adenocarcinoma, not only to develop more effective treatments for gastric cancer, but also because it might serve as a paradigm for the role of chronic inflammation in the genesis of other malignancies that arise within the gastrointestinal tract. PMID- 16362990 TI - Viral gene therapy strategies: from basic science to clinical application. AB - A major impediment to the successful application of gene therapy for the treatment of a range of diseases is not a paucity of therapeutic genes, but the lack of an efficient non-toxic gene delivery system. Having evolved to deliver their genes to target cells, viruses are currently the most effective means of gene delivery and can be manipulated to express therapeutic genes or to replicate specifically in certain cells. Gene therapy is being developed for a range of diseases including inherited monogenic disorders and cardiovascular disease, but it is in the treatment of cancer that this approach has been most evident, resulting in the recent licensing of a gene therapy for the routine treatment of head and neck cancer in China. A variety of virus vectors have been employed to deliver genes to cells to provide either transient (eg adenovirus, vaccinia virus) or permanent (eg retrovirus, adeno-associated virus) transgene expression and each approach has its own advantages and disadvantages. Paramount is the safety of these virus vectors and a greater understanding of the virus-host interaction is key to optimizing the use of these vectors for routine clinical use. Recent developments in the modification of the virus coat allow more targeted approaches and herald the advent of systemic delivery of therapeutic viruses. In the context of cancer, the ability of attenuated viruses to replicate specifically in tumour cells has already yielded some impressive results in clinical trials and bodes well for the future of this approach, particularly when combined with more traditional anti-cancer therapies. PMID- 16362991 TI - Infection and disease: cause and cure. AB - Much can be learnt about the mechanisms by which micro-organisms cause disease from the ways that they interact with cells and tissues. This issue of The Journal of Pathology contains articles that address the roles that cell and tissue biology and pathology are playing in the elucidation of these mechanisms. A review of variant Creutzfeldt-Jakob disease is followed by a discussion of severe acute respiratory syndrome (SARS). Two articles on human papillomavirus (HPV) infection address the association between viral infection and neoplasia, as do reviews on viruses and lymphoma/leukaemia, and Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8, HHV8). The section on viral disease concludes with an article on morbilliviruses. The intracellular effects of bacteria are addressed in a review of Listeria infection and a further review outlines recent advances in our knowledge of syphilis. Reviews on Helicobacter and gastric neoplasia, innate defences against methicillin-resistant Staphylococcus aureus (MRSA) infection, and the function of granulomas in tuberculosis also address aspects of tissue responses to bacterial infection. Following a review of the function of immunoglobulin A in defence against infection, a group of articles considers vaccination and gene therapy approaches, the latter involving consideration of both viral and bacterial strategies. The reviews assembled here bridge several gaps: between microbiology and cellular pathology; between host and infecting organism; and between disease and therapy. It is clear that cell and tissue pathology approaches are of value in all of these spheres, providing cell and tissue relevance to microbiological and immunological observations. PMID- 16362992 TI - How the SARS coronavirus causes disease: host or organism? AB - The previous epidemic of severe acute respiratory syndrome (SARS) has ended. However, many questions concerning how the aetiological agent, the novel SARS coronavirus (CoV), causes illness in humans remain unanswered. The pathology of fatal cases of SARS is dominated by diffuse alveolar damage. Specific histological changes are not detected in other organs. These contrast remarkably with the clinical picture, in which there are apparent manifestations in multiple organs. Both pathogen and host factors are important in the pathogenesis of SARS. The choice of specific receptors and the unique genome of the SARS-CoV are important elements in understanding the biology of the pathogen. For the host cells, the outcome of SARS-CoV infection, whether there are cytopathic effects or not, depends on the cell types that are infected. At the whole-body level, immune mediated damage, due to activation of cytokines and/or chemokines and, perhaps, autoimmunity, may play key roles in the clinical and pathological features of SARS. Continued research is still required to determine the pathogenetic mechanisms involved and to combat this new emerging human infectious disease. PMID- 16362993 TI - Innate defences against methicillin-resistant Staphylococcus aureus (MRSA) infection. AB - The innate immune system is the primary defence against bacterial infection. Among the factors involved in innate defence, anti-microbial peptides produced by humans have recently attracted attention due to their relevance to some diseases and also to the development of new chemotherapeutic agents. Staphylococcus aureus is one of the major human pathogens, causing a variety of infections from suppurative disease to food poisoning. Methicillin-resistant S. aureus (MRSA) is a clinical problem and with the recent emergence of a vancomycin-resistant strain, this will pose serious problems in the near future. In investigating the molecular biology of S. aureus infections to develop new chemotherapeutic agents against MRSA infections, knowledge of the interaction of innate anti-microbial peptides with S. aureus is important. In vitro and in vivo experiments demonstrate that exposure of S. aureus to host cells can induce the anti microbial peptides beta-defensin-2 (hBD2), hBD3, and LL37/CAP18. The induction level of these peptides differs among strains, as does the susceptibility of the strains, with MRSA strains exhibiting lower susceptibility. In summary, the susceptibility of S. aureus strains, including MRSA strains, to components of the innate immune system varies, with the MRSA strains showing more resistance to both innate immune factors and chemotherapeutic agents. PMID- 16362994 TI - HPV-mediated cervical carcinogenesis: concepts and clinical implications. AB - Persistent infection with a high-risk human papillomavirus (hrHPV) is generally accepted as a necessary cause of cervical cancer. However, cervical cancer is a rare complication of an hrHPV infection since most such infections are transient, not even giving rise to cervical lesions. On average, it takes 12-15 years before a persistent hrHPV infection may ultimately, via consecutive premalignant stages (ie CIN lesions), lead to an overt cervical carcinoma. This argues that HPV induced cervical carcinogenesis is multi-step in nature. In this review, the data from hrHPV-mediated in vitro transformation studies and those obtained from analysis of clinical specimens have been merged into a cervical cancer progression model. According to this model, a crucial decision maker in the early stages following infection involves individual susceptibility for certain HPV types depending on the genetic make-up of immune surveillance determinants. Once a CIN lesion has developed, altered transcriptional regulation of the viral E6/E7 oncogenes, resulting in genomic instability and distinguishing the process of cell transformation from a productive viral infection, probably provides the subsequent important step towards malignancy. The additional (epi)genetic alterations that subsequently accumulate in high-grade CIN lesions may result in overt malignancy via immortality and growth conditions that gradually become less sensitive to growth-modulating influences mediated by cytokines and cell-cell and cell-matrix adhesions. The potential implications of hrHPV testing and some other biomarkers deduced from this model for cervical screening and the clinical management of CIN disease are also discussed. PMID- 16362995 TI - HPV-associated skin disease. AB - Human papillomaviruses (HPVs) are DNA tumour viruses that induce hyperproliferative lesions in cutaneous and mucosal epithelia. The relationship between HPV and non-melanoma skin cancer (NMSC) is important clinically since NMSC is the most common form of malignancy among fair-skinned populations. It is well established that solar ultraviolet (UV) irradiation is the major risk factor for developing NMSC, but a pathogenic role for HPV in the development of NMSC has also been proposed. Recent molecular studies reveal a likely role for HPV infection in skin carcinogenesis as a co-factor in association with UV. This review summarizes the literature describing these data, highlights some of the important findings derived from these studies, and speculates on future perspectives. PMID- 16362996 TI - Viruses and lymphoma/leukaemia. AB - Viruses of the retrovirus and herpesvirus families are aetiological agents of human leukaemias and lymphomas. The human T-cell leukaemia virus type 1 causes adult T-cell leukaemia and the Epstein-Barr virus is associated with Burkitt's lymphoma, lymphomas in immunosuppressed people, and Hodgkin lymphoma. The discovery of human herpesvirus type 8 has led to the identification of a rare and unusual group of virus-associated lymphoproliferative diseases. Individuals infected with the human immunodeficiency virus are at greatly increased risk of developing lymphoma but here the mechanism of lymphomagenesis is indirect. Recent data suggest that hepatitis C virus infection is also associated with an increased incidence of lymphoma, whereas data relating to SV40 remain controversial. PMID- 16363001 TI - Care and support needs of children and young people with cancer and their parents. AB - The importance of psychosocial support services for children with cancer and their families is recognised but evaluation of such services is less well developed with little information available about different patterns of provision. This paper provides an overview of psychosocial support children and their families in the UK receive during and after treatment. It reports the results of a postal survey of 303 families, within which parents and children identified their satisfaction with support services and also areas of unmet need. Satisfaction was identified in a range of areas, including medical information and support from nurses and social workers. However, areas of unmet need were also highlighted, especially age appropriate facilities, emotional support and information in different formats. Although British government policy currently seeks to develop standards and guidelines of care throughout the National Health Service, this paper demonstrates that there is still a need to develop psychosocial support services and work towards recently established guidelines in order to ensure that families receive flexible but equitable packages of care and support, wherever treatment is received. PMID- 16362999 TI - Association of coping style, pain, age and depression with fatigue in women with primary breast cancer. AB - The purpose of this study was to explore the relative contributions of coping, depression, pain and age, in the experience of cancer related fatigue. A total of 353 women treated for primary breast cancer were assessed within one year of diagnosis using the Profile of Mood States, the Hospital Anxiety and Depression Scale and the mini-Mental Adjustment to Cancer Scale. Fatigue was positively associated with depression and pain, but inversely related to age. In contrast to our expectations, fighting spirit was not associated with less fatigue. A relationship between coping style and cancer-related fatigue was found exclusively for 'positive reappraisal', a combination of fighting spirit and fatalism. Detectable only in multivariate analysis together with depression, the results suggest a weak association between coping and fatigue. The relationship between cancer related fatigue, age and coping styles requires further exploration within longitudinal studies. PMID- 16363005 TI - Synthesis, crystal structure, and catalytic properties of MgCo6Ge6. AB - The ternary compound MgCo6Ge6 represents a novel member of the RM6X6 phases, which contains a graphite-type Ge network, Kagome nets of Co atoms, and Ge2 dumbbells with an unexpected short Ge-Ge contact in the range of a localized Ge Ge single bond. The title compound shows a large variety of chemical bonding, which ranges from metallic to multicenter and covalent bonding. The role of polar intermetallic alloys as promising candidates for the application as catalysts for the selective hydrogenation of alpha,beta-unsaturated aldehydes is discussed. MgCo6Ge6 possesses a remarkable activity and selectivity for the hydrogenation of cis/trans-citral to geraniol and nerol. PMID- 16363006 TI - Self-assembled, kinetically locked, Ru(II)-based metallomacrocycles: physical, structural, and modeling studies. AB - By using a "complex as ligand approach," the metal-ion-templated self-assembly of heterometallic tetranuclear metallomacrocycles containing kinetically locked Ru(II) centers is described. Depending on the metal-ion template employed in the self-assembly process, the final macrocycle can be kinetically labile or inert. Electrochemical studies reveal that the kinetically inert macrocycles display reversible Ru(III/II) oxidation couples. The crystal structure of a kinetically inert Ru2Re2 macrocycles reveals a structurally complex palmate anion-binding pocket. Host-guest studies carried out with the same macrocyle in organic solvents reveals that the complex functions as a luminescent sensor for anions and that binding affinity and luminescent modulation is dependent on the structural nature and charge of the guest anion. Computational density functional theory (DFT) studies support the hypothesis that the luminescence of the macrocycle is from a 3MLCT state and further suggests that the observed guest induced luminescence changes are most likely due to modulation of nonradiative decay processes. PMID- 16363007 TI - Photophysical studies of anion-induced colorimetric response and amplified fluorescence quenching in dipyrrolylquinoxaline-containing conjugated polymers. AB - The dipyrrolylquinoxaline (DPQ)-containing monomer and polymers were synthesized and employed as chromogenic and fluorescent chemosensors for inorganic anions. We have found that in the presence of fluoride or pyrophosphate, the receptors do not form hydrogen bonds between the pyrrole protons and anions. The colorimetric responses and fluorescence quenching in these chemosensors are indeed the result of deprotonation of the N-H proton. The anion selectivity is primarily determined by the relative basicity of anions. The sensitivity of DPQ-based chemosensor was found to display a 34-fold enhancement by incorporation into the conjugated polymer. The anion-induced deprotonation generates low-energy, non-fluorescent trapping sites and is responsible for the signal amplification where the quenching of the excited state occurs from the deprotonated DPQ site in the network by rapid exciton migration along the polymeric backbone. PMID- 16363008 TI - Novel 2,1,3-benzothiadiazole-based red-fluorescent dyes with enhanced two-photon absorption cross-sections. AB - This paper reports the two-photon absorbing and orange-red fluorescence emitting properties of a series of new 2,1,3-benzothiadiazole (BTD)-based D-pi-A-pi-D-type and star-burst-type fluorescent dyes. In the D-pi-A-pi-D-type dyes 1-6, a central BTD core was connected with two terminal N,N-disubstituted amino groups via various pi-conjugated spacers. The star-burst-type dyes 8 and 10 have a three branched structure composed of a central core (benzene core in 8 and triphenylamine core in 10) and three triphenylamine-containing BTD branches. All the BTD-based dyes displayed intense orange-red color fluorescence in a region of 550-689 nm, which was obtained by single-photon excitation with good fluorescent quantum yield up to 0.98 as well as by two-photon excitation. Large two-photon absorption (TPA) cross-sections (110-800 GM) of these BTD dyes were evaluated by open aperture Z-scan technique with a femtosecond Ti/sapphire laser. The TPA cross-sections of D-pi-A-pi-D-type dyes 2-6 with a benzene, thiophene, ethene, ethyne, and styrene moiety, respectively, as an additional pi-conjugated spacer are about 1.5-2.5 times larger than that of 1c with only a benzene spacer. The TPA cross-sections significantly increased in three-branched star-burst-type BTDs 8 (780 GM) with a benzene core and 10 (800 GM) with a triphenylamine core, which are about 3-5 times larger than those of the corresponding one-dimensional sub units 9 (170 GM) and 11 (230 GM), respectively. The ratios of sigma/e(pi) between three-branched and one-dimensional dyes were 6.5:3.8 (for 8 and 9) and 6.0:4.0 (for 10 and 11), which are larger than those predicted simply on the basis of the chromophore number density (1:1), according to a cooperative enhancement of the two-photon absorbing nature in the three-branched system. PMID- 16363009 TI - Asymmetric hydrogenation of imines and olefins using phosphine-oxazoline iridium complexes as catalysts. AB - Herein we describe the synthesis of a new class of chiral phosphine-oxazolines and their application as ligands in iridium-catalyzed hydrogenations. Mechanistic aspects of olefin hydrogenation with this class of iridium catalysts are discussed and a selectivity model to help rationalize the results obtained is also presented. PMID- 16363010 TI - Neutron and X-ray diffraction and spectroscopic investigations of intramolecular [C-H...F-C] contacts in post-metallocene polyolefin catalysts: modeling weak attractive polymer-ligand interactions. AB - A family of Group 4 post-metallocene catalysts, supported by fluorine functionalized tridentate ligands with the fluorine substituent in the locality of the metal center, is described. For the first time, the contentious C-H...F-C interaction has been characterized by a neutron diffraction study, which has allowed the position of the hydrogen atoms to be accurately determined. The nature of the weak intramolecular C-H...F-C contacts in these complexes in solution and the solid state was probed by using multinuclear NMR spectroscopy in tandem with neutron and X-ray crystallography. Evidence is presented to demonstrate that the spectroscopic C-H...F-C coupling occurs "through-space" rather than "through-bond" or by MF coordination. The titanium catalysts exhibit excellent activities and high co-monomer incorporation in olefin polymerization. The observed intramolecular C-H...F-C interactions are important with regards to potential applications in polyolefin catalysis because they substantiate the proposed ortho-F...H(beta) ligand-(polymer chain) contacts derived from DFT calculations for the remarkable fluorinated phenoxyimine Group 4 catalysts. Compared with agostic and co-catalyst...metal contacts, weak attractive noncovalent interactions between a polymer chain and a judiciously designed "active" ligand is a new concept in polyolefin catalysis. PMID- 16363011 TI - Psychological distress and coping in breast cancer patients and healthy women whose parents survived the Holocaust. AB - Psychological distress levels of breast cancer patients whose parents were Holocaust survivors ('second-generation Holocaust' patients) were previously shown to be significantly higher than those of a matched group of patients with non-traumatized parents. In this study, we investigated whether this effect reflects only the generally higher distress levels of second-generation Holocaust women or whether breast cancer patients with traumatized parents also present lower adaptation abilities, which result in increased distress to the breast cancer diagnosis. We assessed psychological distress and measures of coping in 193 second-generation Holocaust patients diagnosed with breast cancer, 164 breast cancer patients with non-traumatized parents, 176 healthy second-generation Holocaust women, and 143 healthy women with non-traumatized parents. The main effect of cancer and the main effect of second-generation Holocaust survivor on psychological distress were found to be significant. These two factors (cancer x second generation) had a synergistic effect on the levels of depression and psychoticism. These results support the hypothesis that, at least on some psychological measures, the cumulative distressing effect of having traumatized parents and breast cancer diagnosis is higher than the effect of each factor alone. PMID- 16363012 TI - Controllable growth of straight nanorods and nanowires in the Langmuir films of a bolaamphiphilic par derivative. AB - A novel bolaamphiphilic compound, 1, 10-bis[3'-hydroxy-4'-(2'' pyridylazo)phenoloxy] decane [(PAR)2C10], was synthesized and its spreading film and in situ coordination with metal ions in the Langmuir monolayer at the air/water interface were investigated. It was found that (PAR)2C10 could be spread on the water surface, and the Langmuir film showed a phase transition from a flat conformation to a U-shaped conformation upon compression. Interfacial coordination between (PAR)2C10 and metal ions could occur in situ in the Langmuir monolayer. Depending on the concentration of the subphase, different coordination modes were observed. A 2:1 (ligand-PAR-to-metal-ion) complex was formed at a lower concentration of the subphase, while a 1:1 complex was obtained on the subphase with a higher concentration of the salt. Interestingly, very straight nanowires, extending to several micrometers, were observed in the AFM images of the films transferred from the subphase containing CuCl2 in higher concentrations. However, tortuous nanowires were observed under the subphase containing Cu(CH3COO)2. The formation of such kind of wire-like structures was only observed for the combination of the bolaamphiphilic PAR derivative with the Cu(II) ions. Other metal ions or the single-chain PAR derivative did not show this behavior. A possible growth mechanism of the nanowires was proposed based on the coordination of the bolaamphiphilic (PAR)2C10 with the Cu(II) ions as well as with the counteranions. PMID- 16363013 TI - Single-molecule spectroscopy of uniaxially oriented terrylene in polyethylene. AB - Single terrylene molecules doped into linear low-density polyethylene can be oriented by tensile deformation of the matrix. In measurements on ensembles at ambient and on single terrylene molecules at cryogenic temperature, strong orientation along the stretching direction was observed by polarization-resolved confocal microscopy. At cryogenic temperatures narrow and spectrally stable zero phonon lines were found. The low saturation intensity of 0.07 W cm(-2) is consistent with an uniaxial orientation of terrylene in the sample plane. PMID- 16363014 TI - Randomized clinical trial comparing laparoscopic and open surgery for colorectal cancer within an enhanced recovery programme. AB - BACKGROUND: Laparoscopic resection of colorectal cancer may improve short-term outcome without compromising long-term survival or disease control. Recent evidence suggests that the difference between laparoscopic and open surgery may be less significant when perioperative care is optimized within an enhanced recovery programme. This study compared short-term outcomes of laparoscopic and open resection of colorectal cancer within such a programme. METHODS: Between January 2002 and March 2004, 62 patients were randomized on a 2 : 1 basis to receive laparoscopic (n = 43) or open (n = 19) surgery. All were entered into an enhanced recovery programme. Length of hospital stay was the primary endpoint. Secondary outcomes of functional recovery, quality of life and cost were assessed for 3 months after surgery. RESULTS: Demographics of the two groups were similar. Length of hospital stay after laparoscopic resection was 32 (95 per cent confidence interval (c.i.) 7 to 51) per cent shorter than for open resection (P = 0.018). Combined hospital, convalescent and readmission stay was 37 (95 per cent c.i. 10 to 56) per cent shorter (P = 0.012). The relative risk of complications, quality of life results and cost data were similar in the two groups. CONCLUSION: Despite perioperative optimization of open surgery for colorectal cancer, short term outcomes were better following laparoscopic surgery. There was no deterioration in quality of life or increased cost associated with the laparoscopic approach. PMID- 16363015 TI - Structure and dynamics of water confined in dimethyl sulfoxide. AB - We study the structure and dynamics of hydrogen-bonded complexes of H2O/D2O and dimethyl sulfoxide (DMSO) by infrared spectroscopy, NMR spectroscopy and ab initio calculations. We find that single water molecules occur in two configurations. For one half of the water monomers both OH/OD groups form strong hydrogen bonds to DMSO molecules, whereas for the other half only one of the two OH/OD groups is hydrogen-bonded to a solvent molecule. The H-bond strength between water and DMSO is in the order of that in bulk water. NMR deuteron relaxation rates and calculated deuteron quadrupole coupling constants yield rotational correlation times of water. The molecular reorientation of water monomers in DMSO is two-and-a-half times slower than in bulk water. This result can be explained by local structure behavior. PMID- 16363016 TI - Alignment of colloidal CdS nanowires embedded in polymer nanofibers by electrospinning. PMID- 16363017 TI - Sol-gel template synthesis and photoluminescence of n- and p-type semiconductor oxide nanowires. AB - A sol-gel template technique has been put forward to synthesize single crystalline semiconductor oxide nanowires, such as n-type SnO2 and p-type NiO. Scanning electron microscopy and transmission electron microscopy observations show that the oxide nanowires are single-crystal with average diameters in the range of 100-300 nm and lengths of over 10 microm. Photoluminescence (PL) spectra show a PL emission peak at 401 nm for n-type semiconductor SnO2, and a PL emission at 407 nm for p-type semiconductor NiO nanowires, respectively. Correspondingly, the observed violet-light emission at room temperature is attributed to near-band-edge emission for SnO2 nanowires and the 3d(7)4s-->3d8 transition of Ni2+ for NiO nanowires. PMID- 16363018 TI - Formation of oriented polypeptides on Au(111) surface depends on the secondary structure controlled by peptide length. AB - We synthesized three different lengths of poly(L-lysine) containing an -SH group at the terminal (PLL(n)-SH, n (polymerization degree) = 4, 10, 30) and adsorbed them on an Au(111) surface. To analyze the formation process and the structure of self-assembled monolayers (SAMs), we used atomic force microscopy (AFM) and Fourier transform infrared reflection absorption spectra (FT-IR RAS). At the initial stage of SAM growth, formation of nanosize domains was confirmed by AFM imaging. The alpha-helical PLL(30)-SH exhibited a well-defined SAM structure after adsorption reached equilibrium. The alpha-helical PLL(30)-SH was almost perpendicular to the gold surface and exhibited interesting molecular packing due to the secondary structure of PLL(30)-SH and the underlying Au(111) array. The tilt angle of the helix axis from the substrate normal was estimated to be about 50 degrees (AFM) and 44 degrees (FT-IR RAS) respectively. On the other hand, PLL(4)-SH and PLL(10)-SH formed beta-sheet-type SAMs on the Au(111) surface based on the structure determined by FT-IR RAS spectrum. PMID- 16363019 TI - Pathological features as predictors of recurrence after radical resection of gastric cancer. AB - BACKGROUND: The aim of this study was to investigate the pattern and timing of recurrence and to determine associated risk factors after radical resection of gastric cancer including D2 dissection. METHODS: A total of 274 patients who had undergone radical resection of gastric cancer with nodal involvement or T3-4 tumour were randomized to receive chemotherapy or no further treatment (control group). Locoregional recurrence and distant metastasis were analysed in a competing risks framework, by estimating the crude cumulative incidence in each group. Multiple regression models were used to investigate the influence of treatment and pathological features on the risk of recurrence. RESULTS: Overall, the 7 year rate of locoregional relapse was 15.8 per cent and that of distant recurrence was 34.5 per cent. There was a significant association between pathological node (pN) stage and distant relapse (P < 0.001), and between pathological tumour (pT) stage and locoregional recurrence (P = 0.024). Chemotherapy had no significant effect on either locoregional or distant recurrence. CONCLUSION: The rate of locoregional recurrence after radical surgery for gastric cancer was lower than that in studies based on more conservative surgery. The pT stage was related to the rate of locoregional recurrence whereas pN stage had an impact on distant recurrence. PMID- 16363020 TI - Clinical significance of immunohistochemically detectable lymph node metastasis in adenocarcinoma of the ampulla of Vater. AB - BACKGROUND: The aim of this study was to assess the impact of immunohistochemically identified lymph node metastasis on survival in patients with carcinoma of the ampulla of Vater. METHODS: Three hundred and twenty-six regional lymph nodes dissected from pancreatoduodenectomy specimens from 25 patients with ampulla of Vater carcinoma were immunostained with anticytokeratin antibody (CAM 5.2). The clinicopathological significance of immunohistochemically detectable lymph node metastasis was evaluated and compared with that of other potential prognostic factors. RESULTS: The frequency of lymph node involvement in relation to the total number of dissected lymph nodes increased from 5.5 per cent (18 of 326) using haematoxylin and eosin staining to 9.5 per cent (31 of 326) using cytokeratin immunostaining (P < 0.001). Lymph node involvement was revealed by haematoxylin and eosin staining in eight of 25 patients and by cytokeratin immunostaining in 11 of 25 patients (P = 0.006). Absence of immunohistochemically detectable lymph node metastasis was identified as an independent predictor of improved postoperative survival. CONCLUSION: Immunostaining of dissected lymph nodes adds additional information to data obtained by conventional haematoxylin and eosin staining when determining the prognosis of patients with carcinoma of the ampulla of Vater. PMID- 16363021 TI - Randomized clinical trial examining psychosocial and quality of life benefits of bilateral breast reduction surgery. AB - BACKGROUND: The aim was to determine the effects of bilateral reduction mammaplasty on quality of life and psychosocial functioning in women with mammary hypertrophy. METHODS: Seventy-three women who were referred to either the Hull Breast Unit or Hull Plastic and Reconstructive Surgery Unit were randomized to early or delayed surgery. Both groups had quality of life and psychosocial assessment. Each group underwent two sets of tests. Women who had early bilateral breast reduction were tested before and at 4 months after surgery, whereas those in the control group were tested at the time of randomization and 4 months later, before undergoing surgery. RESULTS: All 73 women completed the study. Mean age was 39 years, and the two groups were well matched for age, body mass index and breast dimension. There were highly significant differences between groups in scores measured on the Functional Assessment of Non-Life Threatening Conditions version 4, EuroQoL, and both mental and physical scales of Short Form 36 (P < 0.001). The Eysenck Personality Questionnaire-Revised demonstrated a statistically significant increase in extroversion and emotional stability in the early treatment group. CONCLUSION: Reduction mammaplasty significantly improved quality of life, and increased extroversion and emotional stability. PMID- 16363022 TI - Planetary science: Huygens rediscovers Titan. PMID- 16363023 TI - Circulating endothelial progenitor cells. PMID- 16363024 TI - Circulating endothelial progenitor cells. PMID- 16363025 TI - Circulating endothelial progenitor cells. PMID- 16363026 TI - Asian scientists and the "glass ceiling". PMID- 16363027 TI - Circulating endothelial progenitor cells. PMID- 16363028 TI - Asian scientists and the "glass ceiling". PMID- 16363029 TI - Asian scientists and the "glass ceiling". PMID- 16363030 TI - Asian scientists and the "glass ceiling". PMID- 16363031 TI - A sad day for science at the FDA. PMID- 16363032 TI - A sad day for science at the FDA. PMID- 16363033 TI - In memoriam: Georgeanna Seegar Jones, M.D.: her legacy lives on. PMID- 16363034 TI - [New neurological symptoms following cerebral infarction in a 59-year old woman]. PMID- 16363035 TI - [Not all patients with paralysis have stroke!]. PMID- 16363036 TI - Gray area for new heart failure drug. Although the FDA approved BiDil for blacks with heart failure, it may work in anyone. PMID- 16363037 TI - Don't let decongestants squeeze your heart. As many over-the-counter decongestants get a new ingredient, you might want to look for alternatives. PMID- 16363038 TI - Better prescriptions needed for prescription success. Sticking with a statin or blood pressure medication isn't easy. Some simple strategies can help. PMID- 16363040 TI - Stamp of approval. PMID- 16363039 TI - Can you hold that heart attack until office hours? In the average hospital, artery-opening angioplasty gets started sooner if you have a heart attack during office hours. The delay is relatively small, though, so don't wait until 7 a.m. to get to the hospital. PMID- 16363041 TI - Ask the doctor. Are the omega-3 fats in flaxseed oil as good for the heart as those in fish or fish oil? Do they have the same side effects? The American Heart Association warns about getting more than 3 grams of omega-3 fats a day without a doctor's supervision, but one teaspoon of my concentrated flaxseed oil--the recommended dose--contains 6.2 grams. Should I be worried? PMID- 16363042 TI - Are resistance puzzlers caught in a data quagmire? XIV International HIV Drug Resistance Workshop. June 7-11, 2005, Quebec, Canada. PMID- 16363043 TI - Focus on hepatitis. HIV/HCV coinfection and progression to AIDS. PMID- 16363044 TI - Special coverage of the 43rd IDSA meeting. Texas researchers find a new independent predictor of mortality among HIV patients. Abnormal ECGs have diagnostic potential. AB - A new study shows that electrocardiogram results for HIV patients could show which patients will have a faster disease progression. In the study of 223 HIV positive individuals, those who had abnormal ECG readings had died in within a little more than a decade, while 36 percent of those who had normal or borderline findings had died within the same time frame. PMID- 16363045 TI - Special coverage of the 43rd IDSA meeting. Prisoners with HIV engaged in many risky behaviors before they were imprisoned. AB - HIV-infected people imprisoned in North Carolina had high rates of alcohol use, drug use, and high risk sexual activity within the six-month period before their incarceration, according to a new study. PMID- 16363046 TI - Special coverage of the 43rd IDSA meeting. Self-rated health tool may have prognostic use in HIV treatment. AB - A new study has found that HIV patients who self-rate their health as fair, poor, or bad predict increased mortality several years down the road. PMID- 16363047 TI - Special coverage of the 43rd IDSA meeting. Older HIV patients have higher rates of cell loss, more chronic illnesses. AB - HIV patients under age 40 appear to have fewer non-HIV-related chronic illnesses and have better natural killer (NK) immune cells than HIV patients over age 50, new research suggests. PMID- 16363048 TI - Special coverage of the 43rd IDSA meeting. Rapid HIV testing and counseling proves effective at US-Mexico border. Researchers provide mobile testing services. AB - Mobile HIV testing and counseling using the rapid HIV test proved an effective way to reach an at risk population at the U.S.-Mexico border, a recent study shows. PMID- 16363049 TI - FDA notifications. FDA provides important information for clinicians serving hurricane victims. PMID- 16363050 TI - FDA notifications. Adult treatment guidelines updated. PMID- 16363051 TI - FDA notifications. Accelerated approval of tipranavir. PMID- 16363052 TI - The criminalization of HIV transmission in England and Wales: questions of law and policy. AB - In this article, Matthew Weait and Yusef Azad discuss the current law concerning the criminalization of HIV transmission in England and Wales, and raise some issues about the wider implications of criminalization for those working in the HIV/AIDS sector. The authors look at the way the fault requirement of "recklessness" has been interpreted in the cases. They explore the courts' approach to consent--the defence which those who have appealed against conviction have sought to use. Then the authors raise some questions about the relevance of disclosure and the way the courts have dealt with knowledge about HIV status and the risks associated with unprotected sex. Finally, they discuss the relevance of the nature of the relationship between the accused person and the person to whom HIV has allegedly been transmitted, and touch on the potentially stigmatizing effects that criminalization may have on socio-economically marginalized groups. The authors conclude by discussing some more general policy-related issues. PMID- 16363053 TI - Canadian developments. HIV disclosure no longer required on application form for temporary resident visa. AB - In May 2005, as a result of pressure from advocates, Citizenship and Immigration Canada (CIC) lifted the requirement that short-term visitors to Canada applying for a visa disclose their HIV status on the application form. PMID- 16363054 TI - Canadian developments. Safer tattooing piloted in six federal prisons. AB - In January of 2005, Correctional Service Canada (CSC) began implementing a safer tattooing pilot program in six federal prisons, one women's and five men's institutions. The pilot phase of the program is expected to last until 31 March 2006. PMID- 16363055 TI - Ottawa: City Council approves distribution of crack kits. AB - After a divisive debate that pitted the Medical Officer of Health against the Chief of Police, Ottawa City Council voted in May 2005 to approve the distribution of crack kits to drug users. Similar programs already exist in Toronto, Montreal, Windsor, Guelph and Vancouver. PMID- 16363056 TI - Canadian developments. Bill to export generic drugs comes into force. AB - The Jean Chretien Pledge to Africa Act (the Act), the Canadian legislation aimed at allowing the export of lower-cost medicines to developing countries, came into force on 14 May 2005, exactly one year after it received Royal Assent. PMID- 16363057 TI - Cidofovir incorporation into human keratinocytes with episomal HPV 16 results in nonselective cytotoxicity. AB - OBJECTIVES: Recurrent respiratory papillomatosis (RRP) is caused by human papillomavirus (HPV). Surgical excision is the mainstay of treatment; however, medical therapy including cidofovir, a cytosine analog, has been investigated. Human papillomavirus does not encode a viral DNA polymerase, which is the known target of cidofovir in cytomegalovirus infections. METHODS: In an effort to better understand the usefulness of cidofovir in the treatment of HPV-related disease, we tested cidofovir's ability to inhibit growth, alter gene expression, and inhibit genome replication. RESULTS: With the use of carbon 14-labeled cidofovir in episomal HPV 16-containing keratinocytes, there was a minimal increase in cidofovir incorporation into episomal DNA versus genomic DNA. Cidofovir decreased the copies of episomal HPV 16 in keratinocytes; however, the copies per cell returned to baseline levels once cidofovir was removed. Expression of a viral oncogene (HPV 16 E6) in transformed keratinocytes with episomal HPV 16 was not decreased by cidofovir. Cytotoxicity in head and neck squamous cell carcinoma lines exposed to cidofovir correlated with cell doubling time, and not with HPV status. Also, tonsil keratinocytes transformed with episomal HPV 16 did not exhibit greater cidofovir-mediated toxicity than did telomerase-transformed keratinocytes. CONCLUSIONS: These findings suggest that any potential in vivo benefit of cidofovir therapy results from non-viral specific cell toxicity at the site of application. PMID- 16363058 TI - Influence of vocal fold scarring on phonation: predictions from a finite element model. AB - OBJECTIVES: A systematic study of the influence of vocal fold scarring on phonation was conducted. In particular, phonatory variables such as fundamental frequency, oral acoustic intensity, and phonation threshold pressure (PTP) were investigated as a function of the size and position of the laryngeal scar. METHODS: By means of a finite element model of vocal fold vibration, the viscoelastic properties of both normal and scarred vocal fold mucosae were simulated on the basis of recent rheological data obtained from rabbit and canine models. RESULTS: The study showed that an increase in the viscoelasticity of the scarred mucosa resulted in an increase in fundamental frequency, an increase in PTP, and a decrease in oral acoustic intensity. With regard to positioning of the scar, the PTP increased most significantly when the scar was within +/-2 mm of the superior-medial junction of the vocal folds. CONCLUSIONS: The systematic data obtained in this investigation agree with the general clinical experience. In the future, these findings may be further validated on human subjects as newly emerging technologies such as linear skin rheometry and optical coherence tomography allow the histologic and viscoelastic properties of the normal and scarred vocal fold mucosae to be measured in the clinic. PMID- 16363059 TI - Efficacy of a cochlear implant simultaneous analog stimulation strategy coupled with a monopolar electrode configuration. AB - OBJECTIVES: The present study was performed to evaluate the efficacy and clinical feasibility of using monopolar stimulation with the Clarion Simultaneous Analog Stimulation (SAS) strategy in patients with cochlear implants. METHODS: Speech recognition by 10 Clarion cochlear implant users was evaluated by means of 4 different speech processing strategy/electrode configuration combinations; ie, SAS and Continuous Interleaved Sampling (CIS) strategies were each used with monopolar (MP) and bipolar (BP) electrode configurations. The test measures included consonants, vowels, consonant-nucleus-consonant words, and Hearing in Noise Test sentences with a +10 dB signal-to-noise ratio. Additionally, subjective judgments of sound quality were obtained for each strategy/configuration combination. RESULTS: All subjects but 1 demonstrated open set speech recognition with the SAS/MP combination. The group mean Hearing in Noise Test sentence score for the SAS/MP combination was 31.6% (range, 0% to 92%) correct, as compared to 25.0%, 46.7%, and 37.8% correct for the CIS/BP, CIS/MP, and SAS/BP combinations, respectively. Intersubject variability was high, and there were no significant differences in mean speech recognition scores or mean preference ratings among the 4 strategy/configuration combinations tested. Individually, the best speech recognition performance was with the subject's everyday strategy/configuration combination in 72% of the applicable cases. If the everyday strategy was excluded from the analysis, the subjects performed best with the SAS/MP combination in 37.5% of the remaining cases. CONCLUSIONS: The SAS processing strategy with an MP electrode configuration gave reasonable speech recognition in most subjects, even though subjects had minimal previous experience with this strategy/configuration combination. The SAS/MP combination might be particularly appropriate for patients for whom a full dynamic range of electrical hearing could not be achieved with a BP configuration. PMID- 16363060 TI - [Avian influenza: much ado about nothing]. PMID- 16363061 TI - Notch and NOXA-related pathways in melanoma cells. AB - Notch receptor-mediated intracellular events represent an ancient cell signaling system, and alterations in Notch expression are associated with various malignancies in which Notch may function as an oncogene or less commonly as a tumor suppressor. Notch signaling regulates cell fate decisions in the epidermis, including influencing stem cell dynamics and growth/differentiation control of cells in skin. Because of increasing evidence that the Notch signaling network is deregulated in human malignancies, Notch receptors have become attractive targets for selective killing of malignant cells. Compared with proliferating normal human melanocytes, melanoma cell lines are characterized by markedly enhanced levels of activated Notch-1 receptor. By using a small molecule gamma-secretase inhibitor (GSI) consisting of a tripeptide aldehyde, N-benzyloxycarbonyl-Leu-Leu Nle-CHO, which can block processing and activation of all four different Notch receptors, we identified a specific apoptotic vulnerability in melanoma cells. GSI triggers apoptosis in melanoma cells, but only G2/M growth arrest in melanocytes without subsequent cell death. Moreover, GSI treatment induced a pro apoptotic BH3-only protein, NOXA, in melanoma cells but not in normal melanocytes. The use of GSI to induce NOXA induction overcomes the apoptotic resistance of melanoma cells, which commonly express numerous cell survival proteins such as Mcl-1, Bcl-2, and survivin. Taken together, these results highlight the concept of synthetic lethality in which exposure to GSI, in combination with melanoma cells overexpressing activated Notch receptors, has lethal consequences, producing selective killing of melanoma cells, while sparing normal melanocytes. By identifying signaling pathways that contribute to the transformation of melanoma cells (e.g. Notch signaling), and anti-cancer agents that achieve tumor selectivity (e.g., GSI-induced NOXA), this experimental approach provides a useful framework for future therapeutic strategies in cutaneous oncology. PMID- 16363062 TI - Current view of the role of transforming growth factor beta 1 in skin carcinogenesis. AB - Previously, we have shown that transforming growth factor beta 1 (TGFbeta1) overexpression in suprabasal epidermis suppresses skin carcinogenesis at early stages, but promotes tumor invasion at later stages. To elucidate the role of TGFbeta1 overexpression in naturally occurring human squamous cell carcinomas (SCC), we screened TGFbeta1 expression patterns in human skin SCC samples and found that TGFbeta1 was overexpressed with two distinct patterns: either predominantly in suprabasal layers or throughout tumor epithelia including basal proliferative cells. To determine the effect of TGFbeta1 overexpression in basal keratinocytes, we generated transgenic mice expressing wild-type TGFbeta1 in basal keratinocytes and hair follicles using the K5 promoter (K5.TGFbeta1(wt)). Surprisingly, these mice developed a severe inflammatory skin disorder. Inflammation was also observed in head and neck tissue when TGFbeta1 transgene expression was inducibly expressed in head and neck epithelia in our gene-switch TGFbeta1 transgenic mice. Given the importance of inflammation in cancer development, our data suggest that TGFbeta1-induced inflammation may override its tumor-suppressive effect even at early stages of skin carcinogenesis. This notion is further suggested by our recent study that Smad3 knockout mice were resistant to skin chemical carcinogenesis at least in part via abrogation of endogenous TGFbeta1-induced inflammation. PMID- 16363063 TI - P63 deficiency: a failure of lineage commitment or stem cell maintenance? AB - A critical role for p63 in the development of stratified epithelia, such as the epidermis, has been recognized since the generation of mice lacking p63 expression. The molecular role of p63 in epidermal morphogenesis, however, remained controversial. The epidermal phenotype of p63-/- mice, which are born with a single-layered surface epithelium instead of a fully stratified epidermis, suggested that p63 could have a role in stem cell maintenance or in the commitment to stratification. In this review, we discuss evidence suggesting that p63 is required for the commitment to stratification, making p63 the earliest known gene expressed in the developing epidermis that is specific for the keratinocyte lineage. PMID- 16363064 TI - The tale of the telomere: implications for prevention and treatment of skin cancers. AB - Work in many laboratories over the past decade has established a central role for the telomere in maintaining genomic integrity. Available data may be interpreted to indicate that telomere disruption, whether due to acute DNA damage or progressive telomere shortening, is the initial event that triggers multiple DNA damage responses. The specific initiating event is likely exposure of the otherwise concealed single-stranded 3' overhang, tandem repeats of TTAGGG, a signal that can be provided to cells in the absence of DNA damage by exogenously provided T-oligos. The ability of T-oligo treatment to trigger SOS-like responses and/or to cause selective apoptosis of already malignantly transformed cells may provide an important new means of cancer prevention and treatment. PMID- 16363065 TI - Melanocyte and keratinocyte carcinogenesis: p53 family protein activities and intersecting mRNA expression profiles. AB - Melanocytes and keratinocytes were analyzed for potential roles of p53, p73, and p63 tumor suppressor family proteins and of malignancy-specific gene expression changes in the etiology of multi-step cancer. Melanocytes expressed deltaNp73alpha, two p63 isoforms and p53. Although p21 and Noxa mRNA levels increased following DNA damage, p53 family member binding to p21 and Noxa DNA probes was undetectable, suggesting p53 family-independent responses. In contrast, keratinocytes expressed multiple isoforms each of p73 and p63 that were induced to bind p21 and Noxa DNA probes after ionizing (IR) or after ultraviolet B (UVB) irradiation, correlating with p21 and Noxa mRNA induction and with apoptosis. Interestingly, IR-resistant malignant melanocytes and keratinocytes both exhibited Noxa mRNA induction after UVB treatment, correlating with DNA binding of p53 family proteins to the Noxa probe only in keratinocytes. To uncover other malignancy-specific events, we queried mouse initiated keratinocyte clones for early changes that were exacerbated in malignant derivatives and also differentially expressed in human advanced melanoma versus normal melanocytes. Using a new method for ranking and normalization of microarray data for 5000 probe sets, 27 upregulated and 13 downregulated genes satisfied our query. Of these, the majority was associated with late-stage human cancers and six were novel genes. Thus, clonal lineage mouse models representing early through late cancer progression stages may inform the focus on early, potentially causal events from microarray studies of human cancers, facilitating prognosis and molecular therapy. PMID- 16363066 TI - Imaging melanoma in a murine model using reflectance-mode confocal scanning laser microscopy and polarized light imaging. AB - The light-scattering properties of cutaneous tissues provide optical contrast for imaging the presence and depth of pigmented melanoma in a highly pigmented murine model, the C57/B6 mouse. Early lesions are difficult to identify when viewing black lesions on a black mouse. Two methods were used to image early lesions in this model. (1) A reflectance-mode confocal scanning laser microscope (rCSLM) was built to provide horizontal images (x-y at depth z) and transverse images (x-z at position y) non-invasively in the living mouse. (2) A polarized light imaging (PLI) camera was built using a linearly polarized white light source that viewed the skin through an analyzing linear polarizer oriented either parallel or perpendicular to the illumination's polarization to yield two images, "PAR" and "PER," respectively. The difference image, PAR-PER, eliminated multiply scattered light and yielded an image of the superficial but subsurface tissues based only on photons scattered once or a few times so as to retain their polarization. rCSLM could image melanoma lesions developing below the epidermis. PLI could distinguish superficial from deeper melanoma lesions because the melanin of the superficial lesions attenuated the PAR-PER image, whereas deeper lesions failed to attenuate the PAR-PER image. PMID- 16363067 TI - Pathogenesis and treatment of alcoholic liver disease: progress over the last 50 years. AB - Fifty years ago the dogma prevailed that alcohol was not toxic to the liver and that alcoholic liver disease was exclusively a consequence of nutritional deficiencies. We showed, however, that liver pathology developed even in the absence of malnutrition. This toxicity of alcohol was linked to its metabolism via alcohol dehydrogenase which converts nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide-reduced form (NADH) which contributes to hyperuricemia, hypoglycemia and hepatic steatosis by inhibiting lipid oxidation and promoting lipogenesis. We also discovered a new pathway of ethanol metabolism, the microsomal ethanol oxidizing system (MEOS). The activity of its main enzyme, cytochrome P4502E1 (CYP2E1), and its gene are increased by chronic consumption, resulting in metabolic tolerance to ethanol. CYP2E1 also detoxifies many drugs but occasionally toxic and even carcinogenic metabolites are produced. This activity is also associated with the generation of free radicals with resulting lipid peroxidation and membrane damage as well as depletion of mitochondrial reduced glutathione (GSH) and its ultimate precursor, namely methionine activated to S-adenosylmethionine (SAMe). Its repletion restores liver functions. Administration of polyenylphosphatidylcholine (PPC), a mixture of unsaturated phosphatidylcholines (PC) extracted from soybeans, restores the structure of the membranes and the function of the corresponding enzymes. Ethanol impairs the conversion of beta-carotene to vitamin A and depletes hepatic vitamin A and, when it is given together with vitamin A or beta-carotene, hepatotoxicity is potentiated. Our present therapeutic approach is to reduce excess alcohol consumption by the Brief Intervention technique found to be very successful. We correct hepatic SAMe depletion and supplementation with PPC has some favorable effects on parameters of liver damage which continue to be evaluated. Similarly dilinoleoylphosphatidylcholine (DLPC), PPC's main component, also partially opposes the increase in CYP2E1 by ethanol. Hence, therapy with SAMe +DLPC is now being considered. PMID- 16363068 TI - The effect of dissolved carbon dioxide on cavitation intensity in mechanical heart valves. AB - BACKGROUND AND AIM OF THE STUDY: Mechanical heart valves (MHVs) are known to induce cavitation during closure and rebound. Cavitation may lead to blood element damage and stable bubble formation, with the latter introducing emboli into the cranial circulation and increasing the risk of stroke. Previous research has suggested that CO2 is the primary blood gas involved in stable bubble growth, due to its high solubility compared to that of oxygen or nitrogen. The primary objective of this study is to determine the role that CO2 plays in MHV-induced cavitation bubble formation. METHODS: Degassed water (5 ppm) was supplemented with CO2 at partial pressures of 0, 40 and 100 mmHg. Cavitation was visualized using high-speed videography for 29 mm Bjork-Shiley Monostrut and Medtronic Hall MHVs in the mitral position. Experimental parameters (heart rate, systolic duration, and left ventricular pressure) were adjusted to provide dp/dt values of 500, 2,500 and 4,500 mmHg/s. High-frequency pressure fluctuations of cavitation bubble collapse were detected using a hydrophone. RESULTS: Root-mean square (RMS) values were calculated to quantify the cavitation intensity for both MHVs at the three loading conditions. The images of cavitation bubble formation and collapse were correlated to their respective RMS values. This study revealed no statistical difference between the cavitation intensities produced by either of the MHVs for the range of CO2-supplemented degassed water tested. For example, at the most physiologic loading condition of 2,500 mmHg/s, the RMS values for the Bjork-Shiley Monostrut valve in degassed water containing 0 and 100 mmHg CO2 were 32.7 +/- 3.5 and 34.3 +/- 6.1 mmHg, respectively. CONCLUSION: The results of this in-vitro study show that, despite affecting stable bubble growth, the presence and quantity of dissolved CO2 does not affect the intensity of the cavitation events occurring during impact of the valve occluder with its housing. Therefore, the role of CO2 is limited to stable bubble development. PMID- 16363070 TI - Angina due to coronary spasm soon after coronary stenting. PMID- 16363069 TI - An immunocompromised patient with necrotic chin lesions. PMID- 16363071 TI - Aggressive Staphylococcal pneumonia: from multiple cavities to pneumatocole and giant bullae formation. PMID- 16363072 TI - Who should capture the value of donated tissue? PMID- 16363073 TI - Soliciting organs on the Internet. PMID- 16363074 TI - The politics of embryo transfer. PMID- 16363075 TI - Conservatives, liberals, and medical progress. PMID- 16363076 TI - The moral education of doctors. PMID- 16363077 TI - Mechanical testing of a novel hydrogel nucleus replacement implant. AB - BACKGROUND CONTEXT: Both total disc and nucleus replacement arthroplastic approaches are emerging as viable treatment modalities for discogenic pain resulting from degenerative disc disease. The results of the testing in this investigation show that this novel nucleus replacement implant exhibits physical and mechanical properties which make it uniquely suitable as a replacement to the degenerated nucleus pulposus. PURPOSE: The purpose of this investigation was to evaluate key performance characteristics of a novel nucleus replacement device. STUDY DESIGN: Mechanical tests were performed to evaluate essential properties of the implant. Of key importance was the ability of the implant to withstand fatigue testing and resist extrusion in cadaveric motion segments. Resistance of the implant to radial deformation is also relevant to the ability of the implant to resist extrusion and maintain disc height. METHODS: Resistance to radial bulging was demonstrated by applying axial compressive loads while examining the "footprint" of the implant through a clear plexiglass lower platen. A NeuDisc implant was compared with an isotropic implant produced from the same hydrolyzed polyacrylonitrile hydrogel. Confined compression tests were used to determine "lifting force" when implants were loaded within stainless steel cylinders submerged in Hanks' balanced salt solution. Fatigue tests involved loading for 10 million cycles. Implants were then examined for physical damage, retention of mechanical properties, and for the ability of the device to return to a complete level of hydration. The final evaluation was extrusion testing in a cadaveric model. Implanted specimens were either tested in compression, lateral bending, or flexion until catastrophic failure was apparent. RESULTS: The NeuDisc implant showed considerably less "bulging" when compared with the identically sized isotropic device. Confined compression testing revealed a lifting force of 400 N at 70% hydration for these hydrogel implants. After fatigue testing, all implants were physically intact and possessed similar swelling characteristics to control samples. In cadaver testing, mean force to failure in compression was 3581.3+/ 1558.5 N. Mean moment to failure in lateral bending was 25.6+/-11.1 Nm. In flexion tests, the mean moment to failure was 52.2+/-18.3 Nm. End plate fracture and ligament failure were the most commonly observed failure modes. CONCLUSIONS: These initial results suggest that this novel implant may prove to be a suitable nucleus pulposus substitute. PMID- 16363079 TI - Are all human embryos equal? PMID- 16363078 TI - Don't mention the syndrome. PMID- 16363080 TI - Laws fail to stop India's organ trade. PMID- 16363081 TI - One bad apple: people will never trust science so long as researchers make up results. PMID- 16363082 TI - Victims of genetic discrimination speak up. PMID- 16363083 TI - Tradition, obligation, and healthcare. PMID- 16363084 TI - When does a person begin? PMID- 16363085 TI - Moral status and personal identity: clones, embryos, and future generations. PMID- 16363086 TI - The identity of identity: moral and legal aspects of technological self transformation. PMID- 16363087 TI - Sex-limited genome-wide linkage scan for body mass index in an unselected sample of 933 Australian twin families. AB - Genes involved in pathways regulating body weight may operate differently in men and women. To determine whether sex-limited genes influence the obesity-related phenotype body mass index (BMI), we have conducted a general nonscalar sex limited genome-wide linkage scan using variance components analysis in Mx (Neale, 2002). BMI measurements and genotypic data were available for 2053 Australian female and male adult twins and their siblings from 933 families. Clinical measures of BMI were available for 64.4% of these individuals, while only self reported measures were available for the remaining participants. The mean age of participants was 39.0 years of age (SD 12.1 years). The use of a sex-limited linkage model identified areas on the genome where quantitative trait loci (QTL) effects differ between the sexes, particularly on chromosome 8 and 20, providing us with evidence that some of the genes responsible for BMI may have different effects in men and women. Our highest linkage peak was observed at 12q24 (-log10p = 3.02), which was near the recommended threshold for suggestive linkage (-log10p = 3.13). Previous studies have found evidence for a quantitative trait locus on 12q24 affecting BMI in a wide range of populations, and candidate genes for noninsulin-dependent diabetes mellitus, a consequence of obesity, have also been mapped to this region. We also identified many peaks near a -log10p of 2 (threshold for replicating an existing finding) in many areas across the genome that are within regions previously identified by other studies, as well as in locations that harbor genes known to influence weight regulation. PMID- 16363088 TI - Death or declaw: dealing with moral ambiguity in a veterinary hospital. AB - The medical practice of declawing has received much political debate over the past few years. Yet, empirical and theoretical research on how this practice is maintained and the ethical positions of those who actually participate in this work is lacking. Drawing from 9 months of ethnographic fieldwork in a feline specific veterinary hospital and open-ended interviews with veterinarians and staff, this study examines veterinary staff members' attitudes toward, and strategies for, dealing with the medical practice of declawing. Specifically, findings show that a number of staff felt uncomfortable with their participation in onychectomy (declawing) and relied heavily on organizational support structures to cope both with these feelings and the moral ambiguity about the practice. Relying on these structures, the veterinarians and their staff are able simultaneously to define felines as subjects worthy of respect for their quality of life, protect their own self-identity as people who work toward the best interest of animals, and paradoxically support action toward felines that they find morally objectionable. PMID- 16363089 TI - Universal access to health care and religious basis of human rights. PMID- 16363090 TI - Seeing is believing. PMID- 16363091 TI - Making ES cells 'ethically sound'. AB - Two research groups present alternative approaches for the preparation of embryonic stem (ES) cells that they hope may help bypass the social and political morass surrounding this technology. PMID- 16363092 TI - MicroRNA: the perfect host. AB - By creating an expression cassette that embeds the sequence of short hairpin (sh)RNA into the larger fold of a ubiquitous microRNA, scientists can achieve highly efficient target gene knockdown. PMID- 16363094 TI - Teaching cell biologists how to count. AB - To truly understand the biochemistry of cellular systems, the concentrations of the endogenous protein players must be known, and these can be obtained by careful quantification of fluorescent fusion proteins. PMID- 16363093 TI - The view from Awaji island: past, present, and future of RCC1 and the Ran GTPase system. AB - The International Symposium on Ran and the Cell Cycle was held on October 1-4, 2005, at the Awaji Island Resort near Osaka, to celebrate the career and scientific achievements of Professor Takeharu Nishimoto. One hundred of his former lab members, collaborators and other scientific colleagues from around the world attended the symposium organized by Mary Dasso (National Institutes of Health) and Yoshihiro Yoneda (Osaka University). The program was divided into sessions on cell cycle and chromosomes, nuclear import and export of proteins and RNA, nuclear envelope and the nuclear pore complex, and RCC1 and chromatin. Dr. Nishimoto's retirement from Kyushu University is a perfect time to look back at the history of Ran and RCC1, assess the current state of the field, and discuss the challenges that remain in order to unravel the complexities of the Ran GTPase system. PMID- 16363095 TI - 'Computational evolution' offers riboswitch solution. AB - A computational strategy for the fast and efficient design of sensitive, sequence activated ribozymes could dramatically reduce the time and benchwork investment for scientists interested in engineering specialized RNA-based tools. PMID- 16363096 TI - Decking out your DNA. AB - Certain DNA polymerases show surprising tolerance for chemically modified nucleotides, opening up exciting new possibilities for the efficient production of highly functionalized nucleic acid molecules. PMID- 16363097 TI - A biological tool with chemical applications. AB - Chemistry researchers demonstrate the utility of the sandwich immunoassay for high-throughput screening of catalysts of organic coupling reactions. PMID- 16363098 TI - American Society for Reconstructive Microsurgery (ASRM) annual meeting, Fajardo, Puerto Rico, January 15-18, 2005. Abstracts, part II. PMID- 16363099 TI - The Breast Journal regrettably is retracting a manuscript entitled, " Dolasetron decreases postoperative nausea and vomiting after breast surgery". PMID- 16363100 TI - Sentara builds culture of safety based on proven concepts. PMID- 16363101 TI - Clinical advisor helps PA hospital make improvements. AB - A few years ago, only 40% of the patients at St. Luke's Hospital and Health Network in Bethlehem, PA, who were having an ST-elevation myocardial infarction received angioplasty within 120 minutes. Today, 100% of the patients are getting to the cath lab within the two-hour timeframe. PMID- 16363102 TI - Michigan health plan develops new model of diabetes care. AB - Michigan health plan develops new model of diabetes care. When a physician sees a diabetes patient at one of the centers run by Grand Valley Health Plan, it may not be a one-on-one meeting. Behavioral health counselors and nutritionists are sometimes also in the room. It's important for other health care practitioners to be involved because they can offer advice from another perspective. PMID- 16363103 TI - Lab specimen collection system eliminates label sorting. AB - Phlebotomists at the Jackson-Madison County General Hospital perform more than 18,000 venipunctures a month to collect blood specimens for laboratory tests. Because each of the blood draws involves handling five or six labels for the tubes that the blood is collected in, it means the hospital prints about 90,000 labels a month, which makesfor a lot of label sorting. PMID- 16363104 TI - Colorado hospital creates after hours program for children. AB - It's something that every parent dislikes--their child begins showing signs of an illness after the pediatrician's office closes for the day. Littleton Adventist Hospital has developed a solution that has made treating after-hours pediatric cases easier for everyone. PMID- 16363105 TI - EMR has key role in screening program to reduce MRSA infection. AB - Electronic medical records (EMRs) are playing an important role in a new system wide admission screening program at Evanston Northwestern Healthcare that is designed to detect methicillin-resistant Staphylococcus aureus. PMID- 16363106 TI - Integrating voice recognition and EMR cuts transcription time, costs. AB - A relatively inexpensive speech recognition system integrated with an electronic medical records (EMR) system is saving time and costs at Bronson Methodist Hospital, Kalamazoo, MI. PMID- 16363107 TI - [Intravenous therapy: innovation and safety in nursing]. PMID- 16363108 TI - [Introduction to intravenous therapy]. AB - After defining what intravenous therapy consists of, the author takes his readers through a historical summary about intravenous therapy in order to explain its origins and its applications. The author also describes the principal cases in which intravenous therapy use is warranted. PMID- 16363109 TI - [CDC recommendations: Centers for Disease Control and Prevention of infectious diseases in the United States]. AB - Due to the fact that complications from infections are more frequent and serious related to intravenous therapy it is necessary to implement multidisciplinary measures which reduce their incidence. The recommendations made by the CDC help to guarantee the quality of the services which nursing professionals provides to patients when applying this procedure. PMID- 16363110 TI - [Peripheral access venous catheter]. AB - The author reviews the classifications of different peripheral access venous catheters, analyzing the criteria to select each type of catheter, their indications for use, and the advantages and inconveniences of each one. PMID- 16363111 TI - [Central venous catheters]. AB - In this article the author describes the various systems modem medicine has available to administer intravenous therapy under emergency situations as well as during prolonged use and for chronic patients who need this treatment. PMID- 16363112 TI - [Arterial catheters]. AB - The authors review the fundamental points related to arterial catheters: their applications, how to prepare patients in need of this catheter cleansing and disinfection methods, maintenance and care of these catheters, their installation, how to extract samples with them, how to remove and change these catheters and how to register their use. PMID- 16363113 TI - [Complications caused by intravenous therapy]. AB - Nursing professionals must know everything related to complications caused by intravenous therapy including the ways to prevent and solve these complications. We need not forget that nurses are the ones mainly responsible for the insertion, manipulation, removal and care of catheters. PMID- 16363114 TI - [Pain in vascular canals]. AB - Vascular canalization is a habitual, daily practice for nurses and one which is not exempt from complications such as pain. Therefore, it is necessary that nurses know the factors which bear an influence on the degree of pain patients experience, methods to evaluate pain, as well as how to use clinical guidelines to improve or reduce the variability of pain when employing this technique on patients. PMID- 16363115 TI - [Surgical instruments (IV). A classification of surgical instruments based on their physical traits]. AB - In this last article in a series about surgical instruments, the author continues explaining his classification system according to the degree, branches, cut and consistency instruments have. A bibliography for the entire series of articles accompanies this article. PMID- 16363116 TI - [The elp3 subunit of human elongator complex ignificantly complemented the growth defects of yeast elp3delta strain]. AB - The human elongator complex was found to be very similar to the yeast Elongator both in composition and its interaction with RNA polymerase II. But little is known about its functions in vivo. In this study, we analyzed the functions of the help3, the catalytic subunit of human Elongator, using a yeast complementation system. The results indicated that help3 was able to significantly complement the growth defects of the elp3delta yeast strain under high temperature and caffeine conditions. Gene expression analysis showed that help3 significantly resumed the slow activation of the pho5 gene under the low phosphate condition, and when heat shocked it increased the expression level of ssa3 gene. The yhelp3 containing the HAT domain of human elp3 and the remainder of yeast elp3 had a higher complementation ability than human elp3, while the yhelp3HA T- with the catalytic HAT domain deleted had no complementation ability. These results implicate that human Elp3 subunit had similar functions to those of the yeast, and the HAT activity of human Elp3 was essential for its function. PMID- 16363117 TI - [The character of calcium distribution in developing anther of lettuce (Lactuca sativa L.)]. AB - Potassium antimonite was used to locate calcium in the anther of lettuce (Lactuca sativa L) during its development. At the early stage of anther development there were few calcium granules in microspore mother cells and the cells of anther wall. After meiosis of microspore mother cells, calcium granules first appeared in the tapetal cells in which some small secretive vacuoles containing many calcium granules were formed and secreted into locule. Then, the tapetal cells began to degenerate. At the late stage of microspore, tapetal cells completely degenerated and its protoplast masses moved into anther locule with many calcium granules. Few calcium granules were precipitated in the microspores just being released from tetrad, but some on the surface of exine. Then calcium granules appeared in the nucleus and cytoplasm of early microspores, as wall as in the exine. When microspores formed some small vacuoles containing some calcium granules, and then the small vacuoles fused to form a large vacuole, the calcium granules in the nucleus and cytoplasm evidently decreased, microspore developed to the late stage. The result suggested that calcium is related to the formation of large vacuole in microspores. The wall of microspore also is a main location of calcium granules during its developing. At early microspore some calcium granules began to accumulate in exine, which suggested calcium related with exine formation. At late stage of microspore, most of calcium granules were mainly deposited on the surface of exine. After the first mitosis of microspores, the large vacuole of bicellular pollen disappeared and calcium granules in the large vacuole went back to cytoplasm again. When bicellular pollen synthesized starches some calcium granules appeared on the surface of starches, which suggested calcium may regulate starch synthesis. With amount of starches increasing, calcium granules disappeared from pollen cytoplasm and only some of them located on the surface of pollen. PMID- 16363118 TI - [Ultracytochemical localization of enzymes and DNA analysis during the development of garlic]. AB - It is believed that during storage, the parenchyma cells of garlic (Allium sativum L.) bulb would wither and fade gradually, and nutrients released inside the cells become available for the germination and growth of the young bud. In this study, The distributions of acid phosphatase (APase) and Adenosine Triphosphatase (ATPase) during germination were analyzed based on the method of lead precipitation at the electron microscopic level. It was found that their activities presenting in plasma membrane, cell wall and plasmodesma increased along the different developmental periods during storage. The fact that the most intensive enzymatic activity of APase and ATPase appeared at germination indicates that degradation, transformation and exportation of cell matrix are helpful in complete translocation of nutrient to new bud. The DNA in the degrading parenchyma cells was analyzed using the agarose electrophoresis. Results clearly showed a typical DNA ladder on the gel, indicating that gene controlled, programmed cell death may contribute to the degradation of garlic parenchyma cells. PMID- 16363119 TI - [The cloning and expression analysis of Sox9b in Cyprinus carpio]. AB - Sox9 gene is one of the important transcription factors involved in developments of many tissues and organs, particularly in sex determination and chondrogenesis. In this article, the genomic DNA of Cyprinus carpio were amplified using degenerate primers. It was found for the first time that there were two versions of Sox9 in C. carpio, Sox9a and Sox9b with the same deduced amino acid sequences in the HMG box. There is one intron in this box with the length 704bp in Sox9a and 616bp in Sox9b respectively. Furthermore, a two-phase rapid amplification of cDNA ends(RACE) was used for the isolation of the full length cDNA of Sox9b. Sequence analysis revealed a 2447bp cDNA encoding a protein of 428 amino acid. HMG box of 79-amino-acid motif was confirmed to be from 96 to 174. Sequence alignment exhibited the identity rate of amino acid of Sox9b among ten kinds of animals including C. carpio, which is above 75%. The result indicated the Sox9 gene is very conservative in the progress of evolution. The tissue-specific expression of Sox9b was studied with semi-quantity RT-PCR. The result showed that this gene expresses widespread in several organs of adult C. carpio, with the level of expression in brain and testis higher than any others. PMID- 16363120 TI - [Factors affecting liposome-mediated gene transfection of mouse somatic cells]. AB - We studied the effects of the amount of liposome and plasmid, exposure time of cells to the liposome-plasmid complexes, number of cell passages and cell types on GFP gene transfection of mouse somatic cells. The maximal GFP transgene expression (30.7%) was achieved when mouse fetal fibroblast cells (MFFC) at 70% 90% confluence of passage 3 were exposed for 6 h to the complexes of 4 microg liposome (LipofectAMINE) and 0.3 microg plasmid (pEGFP-N1). Under these conditions, we compared the effect of the number (from primary to 15) of passages on the transfection efficiency of MFFC. The transfection efficiency of MFFC was 10.0%, 28.9% and 7.2% at the primary, 3rd and 15th passage, respectively, which indicated that the transfection efficiency decreased with passaging. When MFFC, mouse oviductal epithelial cells (MOEC) and mouse granulosa cells (MGC) were transfected at passage 3, the transfection efficiency was 27.8%, 13.7% and 14.2%, respectively, under the described transfection conditions. When the cell cycle stages of different cell types at transfection were examined, it was found that 17.2% of MFFC, 8.7% of MOEC and 9.9% of MGC were at M phases of the cell cycle. Examination of the cell cycle stages of MFFC at different passages showed that MFFC at the third passage had the highest percentage of M cells and the percentage decreased afterwards. This suggested that the transfection efficiency was correlated with the percentages of cells at M phase, and provided essential data for improvement of the transfection efficiency. PMID- 16363121 TI - [Transient expression of foreign ble gene in Dunaliella salina]. AB - Plasmid pSP124S with ble gene was transformed into D. salina cell by electroporation. The retention and expression of foreign gene in D.salina was explored, proper parameters were established. A large amount of foreign plasmid was found to be introduced into cells. The plasmid was degraded gradually, but can be detected within 96 hours. The ble gene was efficiently transcribed from foreign promoter and transcription was maintained at least 72 hours. The ble gene can be translated correctly and so can be used as a selectable marker for the research of genetic tranformation in D. salina. PMID- 16363122 TI - [Comparing studies on the differences of CCl4 liver injury and immunity liver injury mice models]. AB - Comparison between CCl4 liver injury model and immunity liver injury model was studied through physiological, biochemical and gene expression profile methods, and the pathological mechanism of them were discussed. There are significant changes about the physiological, biochemical parameters including ALT,AST between two model groups and normal control group. Among the 14112 target genes, 379 and 293 differentially expressed genes were screened out from the both two model groups respectively in gene expression profile experiments, including 105 common, differentially expressed genes. The results indicate that the two models of CCl4 and immunity have some similarities. However, the differences between them are also obvious. It is useful for the further investigation on the liver-injury mechanism and pathological process of the two different liver-injury models. PMID- 16363123 TI - [Effect of glutamine on the expression of grp75 in PC12 cells]. AB - Glutamine (Gln), a conditionally essential amino acid, can be a potential enhancer of the heat stress response. And glucose-regulated protein 75(grp75) is a member of the hsp family. To evaluate the effect of glutamine on the expression of grp75, PC12 cell was cultured with DMEM, glucose-free DMEM, DMEM within Gln, and glucose-free DMEM within Gln, and the expression of grp75 was detected by immunocytochemistry and western blot, the mRNA level of grp75 in the cells was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Data indicated that Gln can upregulated the expression of grp75 in PC12 cell line, and the effect is more significantly in PC12 cell which was glucose deprivation than the normal cell. To investigate the effect of Gln on PC12 cells under glucose deprivation, MTT method was used to monitor the cell viability after Gln treatment for the cell under glucose depriving. The experiments results showed that glutamine at a concentration range of 0.2-40 mmol/L significantly enhanced the cell viability in glucose-free DMEM. And the protective effect to grp75 low expresssion on PC12 cells is markedly decreased. PMID- 16363124 TI - Rat oocytes in early tertiary follicles can attain appropriate maturation in combination culture system. AB - We have developed a "combination culture system", composed of primary differentiated granulosa cells, proestrus anterior pituitary lobe and delta-4 androstenedione. This cultured system effectively induce the maturation of oocytes from early tertiary follicles of diethylstilbestrol (DES)-treated rats. After DES stimulation, an average of 189 oocytes were obtained from early tertiary follicles of unilateral ovaries; 78% of these oocytes, when cultured in this culture system, extruded the first polar body, a criterion of oocyte maturation. Of the mature oocytes, the rate of normal fertilization and egg cleavage were 88% and 93%, respectively. After 96 h of culturing in vitro, 59% of the zygotes developed into morulae or blastocysts. Embryos implanted at the two cell stage were able to develop into healthy individuals. Thus the combination culture system provides an experimental model in which the molecular mechanisms of egg maturation, fertilization, and embryonic development can be studied. PMID- 16363126 TI - [Study on the apoptosis and its mechanism of lovo human colorectal cancer cells induced by SFPS]. AB - Human colorectal cancer (lovo) cells were chose to study the anti-tumor effects of SFPS and explore the significance of caspase 3 in the apoptosis of lovo cells induced by SFPS. Inhibition of the cell proliferation was measured by MTT assay. SFPS induced apoptosis of lovo cells was observed by electron microscopy, flow cytometry and DNA electrophoresis. Furthermore, the expressions of caspase 3 mRNA and pro-caspase 3 were tested by RT-PCR and Western blot. SFPS exhibited anti proliferative activity in the dosage and time-dependent manner. After incubation for 24, 36, 48 and 72 h, the IC50 of SFPS on lovo cells was 375 mg/L, 355 mg/L, 178 mg/L and 60 mg/L, respectively. DNA ladders of lovo cells were showed on agarose gel electrophoresis and the fragments of DNA were integral of 180-200 bp 24 hours after SFPS treatment at the doses of 5 mg/L, 50 mg/L and 300 mg/L. However, mistiness DNA ladder or smear was found in lovo cells treated with 500 mg/L SFPS. When SFPS concentration was 1000 mg/l, DNA ladder disappeared and showed smear entirely. Morphological examination showed chromosomal condensation, karyotheca margination, cell shrinkage and the presence of apoptosis bodies with electron microscopy. With concentration dependent manner, the apoptosis and sub G1 peaks were observed through flow cytometry, but the cell cycle didn't change obviously. Furthermore, the expression of pro-caspase 3 was decreased and the level of caspase 3 mRNA was increased in the time and dose-dependent manner. It suggests that SFPS may induce the apoptosis of lovo cells in vitro resulting in the inhibition of proliferation. And caspase 3 activation may participate in the processes of the apoptosis of lovo cells induced by SFPS. PMID- 16363125 TI - [The study on the construction of expression plasmid containing rHO-1 cDNA, analysis of rHO-1 activity and function of anti-apoptosis in HUVEC]. AB - Xenograft rejection are due to the activation of endothelial cells and the expression of a series of proinflammatory genes encoding adhesion molecules, cytokines/chemokines and procoagulant molecules,leading to endothelial cell injury and apoptosis. HO-1 which acts as a cytoprotective gene can suppress a variety of inflammatory responses to prevent graft rejection. In this study, The plasmid containing HO-1 cDNA was constructed and transfected into human vascular endothelial cells (HUVEC) for expression using DOTAP lipsomal transfection reagents. Cells were homogenized in cell culture lysis reagent (CCLR) and the activity of HO-1 was measured. The apoptosis of HUVEC induced by tumor necrosis factor (TNF)-alpha was analyzed by flow cytometry. Meanwhile, Heme and Sn protoporphyrin (SnPP) were added respectively to evaluate the apoptosis rate of HUVEC. The results showed that the expression of HO-1 gene can be significantly up-regulated in HUVEC transfected with pcDNA3HO1. The apoptosis rate of cells treated with Heme was less than 20% but significantly increased when cells treated with SnPP, the maximum arrived at 95%. The apoptosis rate in heme induced cells was 5-20 times lower than that in SnPP inhibited cells. The apoptosis rate was a negative relation to HO-1 expression. HO-1 protein can inhibit apoptosis in HUVEC. The results provide evidence to support the essential role of HO-1 in the cytoprotective function. PMID- 16363127 TI - [The comparison study on the preparation of chromosomes of the freshwater prawn Macrobrachraium nipponense (Crustacea, Decapoda)]. AB - The Chromosomes of Macrobrachium nipponense had been prepared using the materials of testis or embryo with the chromosome low-osmosis method or cell low-osmosis method. After 5.5 h, the bivalents at meiotic diplotene were appeared. The comparison of the two methods for two different materials showed that the morpha of the chromosome of testis using the low-osmosis method was better than that of using the cell low-osmosis method. Contrarily, the low-osmosis was better than the cell low-osmosis for the morpha of the chromosome of embryo. The bivalents at meiotic diplotene were rod-shaped, and centromere position could be identified easily at that time. The diploid chromosomes numbers were 104. The chromosome formula was N=11M+26SM+4ST+11T. The technique of the chromosome preparation of Macrobrachium nipponense was preliminarily discussed as well. PMID- 16363128 TI - [Polymethyleneamine alkaloids of animal origin. I. Metabolites of marine and microbial organisms]. AB - This review, issued in two parts, describes the information on the structure and biological activity of animal alkaloids derived from polymethyleneamines and produced by marine organisms, wasps, spiders, and microorganisms. Animal alkaloids are outstanding models for developing methods and drugs for the treatment of many human diseases. In the first part, we consider compounds produced by marine and microbial organisms. Some promising synthetic analogues of these alkaloids are used in developing modem preparations for the chelate therapy of excessive blood iron content and antituberculosis, antiproliferative, and immunosuppressive drugs. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363129 TI - [Properties and functions of a new nucleolar protein, Surf-6, in 3T3 mouse cells]. AB - The localization of the specific protein Surf-6 from nucleoli of eukaryotic cells in mitosis and its sensitivity to the treatment of cells with RNase A and DNase I in situ were studied. It was shown that, in interphase nucleoli of 3T3 mouse cells, Surf-6 is probably associated with RNA and practically is not associated with DNA. In mitosis, Surf-6 appears in forming nucleoli after the known RNA binding proteins fibrillarin and B23/nucleofozmin, which are involved in the early and late stages of the assembly of ribosomal particles, respectively. These observations and the regularities of migration of early and late proteins of ribosome assembly to nucleoli in the telophase of mitosis led us to the presumption that Surf-6 is involved in the terminal stages of the assembly of ribosomal particles in murine cells. An immunoblot analysis of the Surf-6 content in synchronized 3T3 cells showed for the first time that Surf-6 is present at all stages of the cell cycle but its content markedly decreases when cells enter the G0 period. Conversely, the activation of cells for proliferation is accompanied by an increase in the Surf-6 content. These observations allow one to regard Surf 6 as a marker of the cell proliferative state and suggest its implication in the regulation of the cell cycle. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363130 TI - [Peptide synthesis in organic media with the use of subtilisin 72 immobilized on a poly(vinyl alcohol) cryogel]. AB - Subtilisin 72 serine protease (EC 3.4.21.14) immobilized on a poly(vinyl alcohol) cryogel was used as a catalyst in the syntheses of N-protected peptide p nitroanilides of the general formulas Z(or Boc)-Xaa-Phe-pNA (Xaa = Leu or Ala), Z Ala-Xaa-Yaa-pNA (Xaa = Leu or Ala; Yaa = Leu or Phe), and Z-Ala-Ala-Xaa-Yaa-pNA (Xaa = Leu, Arg, or Gly; Yaa = Phe, Leu, Gly, Asp, or Glu). The syntheses were carried out in DMF-acetonitrile mixtures. A number of protected di-, tri-, and tetrapeptides were prepared in yields up to 99%. The syntheses were found to retain stereoselectivity under the conditions studied. The activation of carboxyl group of the acylating component was shown to have a positive effect upon the coupling rate. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363131 TI - [Effect of chemical modification of lipase on the regulation of its lipolytic activity in reversed micelles]. AB - Hydrophilized and hydrophobized forms of the lipase from Mucor miehei were obtained by its chemical modification with cellobiose and N-hydroxysuccinimidyl palmitate with a modification degree of 4 in both cases. A comparative analysis of the regulation of the catalytic activities of the native and modified lipases was carried out in the system of reversed micelles of OT aerosol (AOT) in isooctane. The level of catalytic activity of all the lipase preparations in the micellar medium was found to be higher than that in aqueous solution. The chemical modification of lipase did not result in a change in the regulation of the oligomeric composition of the enzyme controlled by the degree of micelle hydration omega0 (micelle size). The kcat dependences on omega0 for each lipase preparation exhibit two maxima, corresponding to the functioning of lipase monomers and tetramers. The changes in the hydrophilic-lipophilic balance of the lipase surface significantly affect the character of the regulation of enzyme activity due to changes in the surfactant concentration (the number of micelles). The lipase hydrophobization results in a decrease in the enzyme activation effect with an increase in the AOT concentration in comparison with the native lipase. The lipase hydrophilization dramatically decreases the activity of lipase tetramer when the AOT concentration is increased. The catalytic activity of the monomer of hydrophilized lipase is practically independent of the AOT concentration. Kinetic data indicate a mixed type of activation of both oligomeric forms of the native and the hydrophobized lipase by AOT molecules and the noncompetitive type of the activation and AOT inhibition of the monomer and the tetramer of the hydrophilized lipase, respectively. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363132 TI - [L-Glutamic acid modulates the cytotoxic effect of tumor necrosis factor in the HL-60 cell line]. AB - L-Glutamic acid was shown to increase the stability of cells of the HL-60 line of human promyelocyte leukemia to the cytotoxic action of tumor necrosis factor alpha (TNF-alpha) due to the inhibition of apoptotic and NF-kappaB-activating cascades induced by this cytokine. At the same time, L-glutamic acid increases the TNF-alpha-mediated differentiating signal and the accompanying enhancement of the phosphatidylinositol-specific phospholipase C activity. Therefore, it is a promising agent for the reduction of total toxicity and inflammatory processes during treatment with TNF-alpha. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363133 TI - [Polymethylene derivatives of nucleic bases with omega-functional groups. V. Pyrimidine- and purine-containing gamma-butyrophenones]. AB - New polymethylene derivatives of nucleic bases containing a keto function in the omega-position were synthesized by alkylation of nucleic bases with 2-(3 chloropropyl)-2-phenyl-1,3-dioxolane and the subsequent deblocking of the keto group; their physicochemical properties were studied. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363134 TI - [Synthesis and antiviral activity of new 5-substituted 2'-deoxyuridine derivatives]. AB - New 5-azole- and 5-oxime-substituted analogues of 2'-deoxyuridine are synthesized. The analogues with azole ring manifest low toxicities and antiherpetic activities on Vero cell culture, the imidazole derivative being the most active. The inhibitory effects of oximes of 5-formyl-deoxyuridine are comparable with those of the azole-containing nucleoside analogues, although their cytotoxicities are found to be higher; oxime of 5-formyldeoxyuridine is particularly toxic. The nucleoside analogues synthesized exhibit no marked activity on cell cultures infected with various variants of poxvirus. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363135 TI - [Enzymatic synthesis of bis(5'-nucleosidyl) tetra- or triphosphates]. AB - The total fraction of aminoacyl-tRNA synthases from Escherichia coli has been shown to catalyze the synthesis of the bis(5'-nucleosidyl) oligophosphates Ap4AZT, Ap4d4T, Ap43TC, and Ap4ACV, as well as Ap3AZT and Ap3d4T, from [alpha 32P]ATP and the corresponding nucleoside-5'-tri(or di)phosphate. The resulting compounds, characterized by HPLC, are resistant to alkaline phosphatase. Ap4AZT, Ap4d4T, and Ap43TC are formed with approximately equal efficiency, whereas the efficiencies of the synthesis of Ap4ACV, Ap3AZT, and Ap3d4T are three- to fivefold lower. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363136 TI - [Application of the duplex-specific nuclease preference method to the analysis of point mutations in human genes]. AB - A new modification of the single nucleotide polymorphism (SNP) analysis (DSNP, duplex-specific nuclease preference) method using the duplex-specific nuclease from the king crab was proposed. The method was used to study SNPs in the following human genes: kRAS, nRAS, hRAS, and p53, the genes of blood coagulation factor V, methyltetrahydrofolate reductase, prothrombin, and apolipoprotein E and a deletion in the BRCA1 gene. DSNP was shown to be useful for the estimation of the mutant allele content in DNA samples. A system for the simultaneous identification of several adjacent single-nucleotide polymorphisms in the kRAS gene was proposed. The approaches could be used to develop test systems for the detection of SNPs in human genes. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363137 TI - [Synthesis and protective activity of beta-glycosides of N-acetylmuramyl-L-alanyl D-isoglutamine with alkylalicyclic and arylaliphatic aglycons]. AB - The following glycosides of N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP) were synthesized: beta-4-tert-butylcyclohexyl MDP, beta-2-(adamant-1-yl)ethyl MDP, beta-2,2-diphenylethyl MDP, and 3-2-(p-biphenyl)ethyl MDP. The starting peracetylated beta-N-acetylglucosaminides were prepared by the oxazoline method. They were converted into 4,6-O-isopropylidene-N-acetyl-D-muramic acids, which were coupled with L-Ala-D-Glu(NH2)OBn. The target glycopeptides were obtained after their deprotection. The stimulation of the antiinfection resistance of mice against Staphylococcus aureus by the MDP glycosides was studied. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363138 TI - [New charge-deficient agmatine analogs]. AB - N,N'-Di-Boc-N"-triflylguanidine was demonstrated to be an efficient guanidinylation reagent for O-substituted hydroxylamines. N-(3-Aminooxypropyl)- and N-(3-aminopropoxy)guanidines, previously unknown isosteric and charge deficient agmatine analogues, have been synthesized. The possibilities of using these compounds in studying polyamine metabolism are discussed. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363139 TI - [Isolation and characteristic of super long-chain unsaturated aldehydes from the freshwater sponge Lubomirskia baicalensis]. AB - Several unusual overlong-chain unsaturated aldehydes (22:1, 22:2, 23:1, 24:1, 24:2, and 25:2) were found in total lipids of the endemic sponge Lubomirskia baicalensis from Baikal Lake. Tetracos-17-enal was identified as the major aldehyde of the mixture using GC-MS and 1H and 13C NMR spectroscopy. A procedure for the isolation of total overlong-chain aldehydes was suggested. We think that the overlong-chain aldehydes defend the sponge from fouling and predators. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363140 TI - [Dipentaerythritol in the synthesis of the new type of phospholipids]. AB - The first representatives of phospholipids of new types were synthesized on the basis of dipentaerythritol, 2,2'-[oxybis(methylen)]bis(hydroxymethyl))-1,3 propandiol. The starting polyol was phosphorylated with hexaethylphosphoric triamide to biphosphite, which was converted into thio- and selenophosphates by treatment with sulfur or selenium. The phosphoacetals were transformed into thio- and selenoanalogues of phospholipids by direct acylation with higher fatty acid chlorides. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363141 TI - [Geometrical configuration of 12,13-epoxy-9,11-octadecadienoic acid, a product of the reaction catalyzed by flaxseed allene oxide synthase (CYP74A)]. AB - The geometrical configuration of a short-living allene oxide reaction product that arises under the catalysis by flaxseed allene oxide synthase (CYP74A) was studied by NMR spectroscopy. The structure of (9Z,11E)-12,13-epoxyoctadeca-9,11 dienoic acid was established for it from the results of the nuclear Overhauser effect. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. PMID- 16363142 TI - Developmental dynamics of some blood biochemical parameters in the growing turkey (Meleagris gallopavo). AB - Blood serum clinical biochemical parameters of fasted BUT Big 8 male turkeys were determined at the ages of 3 days, 4, 8, 12, 16 and 20 weeks, for a follow-up of the developmental changes of some serum metabolites, enzymes and ions. The serum protein content (total protein, albumin, globulin) increased with age, indicating also the moulting-associated metabolic changes in the age interval from the 8th to the 12th weeks. Creatinine was shown to have a peak at 3 days of age (role of muscle activity in thermogenesis), while urate concentration sensitively reflected the dietary protein amount. Serum triglycerides peaked at the time of yolk catabolism, while cholesterol was shown to indicate the moulting, as was serum malondialdehyde. Serum sodium content increased throughout the study. Alanine aminotransferase and aspartate aminotransferase activities increased along the ontogeny, while alkaline phosphatase activity decreased in parallel with the growth. Serum creatine kinase activity showed an over one-magnitude increase. General metabolic and enzymatic alterations were characteristic and applicable for the description of the ontogenetic development of a precocial (post-hatch triglyceride peak), large bodied, meat-type (lactate dehydrogenase, continuously increasing creatine kinase) bird species. PMID- 16363143 TI - In sacco studies of conjugated linoleic acid production from various oils in the rumen of sheep. AB - In this experiment sunflower oil, soybean oil and fish oil were incubated in rumen-fistulated adult ewes (n = 5) to study conjugated linoleic acid (CLA) production in the rumen. The individual oils were incubated in nylon bags in the rumen on perlite carrier (40% oil, 60% carrier) over a period of 2, 4, 6, 8, 10 and 12 h for all treatments. During the incubation of each oil primarily the formation of the cis-9, trans-11 isomer of CLA could be observed. Both sunflower and soybean oils showed similar changes in the rumen. After the incubation of these two vegetable oils the proportion of linoleic acid decreased significantly as the duration of incubation increased in the rumen. These changes were accompanied by a significant increase in the amount of cis-9, trans-11 CLA. However, in the case of sunflower oil the rate of formation of the cis-9, trans 11 CLA isomer was significantly higher after the different incubation times as compared to soybean oil. Much lower amounts of CLA were formed when fish oil was incubated in the rumen. The level of cis-9, trans-11 isomer produced during these treatments was 10% less than the amount obtained with the other two oils of vegetable origin. Besides the cis-9, trans-11 isomer, trans-10, cis-12 CLA could also be detected during the incubation of the different oils in the rumen. However, the level of this isomer was low and did not show consistent differences among the treatments. The results of this experiment indicate that the fatty acid composition of the oils and the duration of incubation collectively determine the amount of CLA produced in the first compartment of the forestomach of ruminants. PMID- 16363144 TI - Effect of alpha-tocopherol on antioxidant enzyme activities and lipid peroxidation in rainbow trout (Oncorhynchus mykiss). AB - In this study, the effect of alpha-tocopherol on the antioxidant capacity of rainbow trout (Oncorhynchus mykiss) was determined. For this purpose, activities of antioxidant enzymes such as catalase (CAT), peroxidase (POD) and glutathione reductase (GSSG-Rx) were investigated. Enzyme activities were measured at 0 (control), 1, 3 and 5 h after alpha-tocopherol injection. In addition, the levels of malondialdehyde (MDA) as a lipid peroxidation marker were determined in the erythrocytes. The results showed that alpha-tocopherol significantly activated the CAT, POD and GSSG-Rx enzymes as compared with the enzyme activities found in the controls (p < 0.05). However, MDA levels were significantly decreased by alpha-tocopherol treatment (p < 0.05). The results suggest that alpha-tocopherol may have a pro-oxidant tendency at a high dose and cause mild oxidative stress which could modulate signal transduction cascades, redirect gene expression, and influence many cellular responses such as proliferation, differentiation, and reproduction. For this reason, alpha-tocopherol should be used carefully in all applications in relation to fish. PMID- 16363145 TI - Study of the energy and protein balance of pregnant rabbit does using two comparative methods. AB - The total body composition of 4 x 10 female, 5-month-old New Zealand White rabbits was determined, using direct chemical analysis and computerised tomography (CT). Ten non-pregnant rabbits (control) and 3 x 10 pregnant rabbits at days 14, 21 and 28 of gestation, respectively, were included in the study. With the advancement of pregnancy, the body weight (BW) of does significantly (P < 0.05) increased (control: 3255 g, day 28: 3938 g), while the dry matter, DM (control: 43.02%, day 28: 40.94%), ether extract, EE (control: 20.15%, day 28: 18.13%), crude protein, CP (control: 20.15%, day 28: 18.13%), and energy concentration decreased (control: 12.41 MJ/kg BW, day 28: 11.81 MJ/kg BW). During pregnancy the protein content of uteri (control: 12.24%, day 28: 49.45% of DM) and the retained energy in the form of uterus with fetuses (day 14: 0.824 MJ, day 28: 2.189 MJ) increased, while the dry matter (control: 82.39%, day 28: 28.96%) and ether extract content (control: 86.05%, day 28: 45.92% of DM) of uteri decreased. With the advancement of pregnancy the dry matter (day 14: 39.70%, day 28: 18.65%) and crude protein (day 14: 75.74%; day 28: 63.63% of DM) content and the number of fetuses (day 14: 8.8; day 28: 6.4) decreased, while their fat concentration (day 14: 10.13%, day 28: 24.34% of DM) increased. According to the equations created from the CT pictures, body fat (R2 = 0.871) and energy (R2 = 0.926) content can be predicted with high accuracy. The accuracy of predicting body protein content is much lower (R2 = 0.356), which can be improved by introducing the body weight as a new dependent variable (R2 = 0.797). It can also be concluded that, in contrast to other animal species, rabbit fetuses show considerable development not only in the last trimester of gestation but already from the second half of pregnancy. PMID- 16363146 TI - Effect of age, sex and season on the prevalence and intensity of helminth infections in domestic pigeons (Columba livia) from Bursa Province, Turkey. AB - The one-year study reported in this paper was carried out from April 2002 to March 2003 in domestic pigeons (Columba livia) of Bursa Province (Turkey) to determine the occurrence of helminth species and to evaluate the effect of host age, sex and season on their prevalence and intensity. A total of 100 (31 young and 69 adult) domestic pigeons were necropsied and examined. The results of necropsies revealed that 74 pigeons harboured helminth infections. Three helminth species were identified: Baruscapillaria obsignata (63%), Ascaridia columbae (42%) and Raillietina echinobothrida (1%). There were no significant differences in the prevalence of B. obsignata and A. columbae by host age and sex. Significantly the highest prevalence rate of A. columbae was observed in the autumn months, but there was no significant difference in the seasonal prevalence of B. obsignata. The mean intensity of B. obsignata and A. columbae was higher in adults than in young pigeons but it did not differ significantly between the two sexes. The highest mean intensity rate of B. obsignata was observed in the autumn, while there was no significant difference for A. columbae with regard to season. From the data obtained in this study it can be concluded that in regions with similar climatic conditions as those in the present study area, pigeon breeders should be cautioned especially of B. obsignata and A. columbae infections of adult pigeons in the autumn months. PMID- 16363147 TI - Comparison of leptin levels in serum and follicular fluid during the oestrous cycle in cows. AB - Leptin is mainly synthesised in white adipose tissue. Besides its effects on body weight and metabolic homeostasis, leptin also has effects on puberty, sexual maturation and reproduction. In this study the relationship between leptin, IGF 1, oestradiol (E2) and progesterone levels were investigated in serum and follicular fluid from cows. This study included 72 healthy, Brown Swiss cows aged 4-5 years. Samples from the jugular vein and follicular fluids were collected. Phases of the oestrus cycle of cows were classified according to their serum progesterone levels (< 3.18 nmol/l, follicular phase and the others as luteal phase). Follicles were grouped as large (> or = 8 mm) or small (< 8 mm). Leptin, IGF-1, oestradiol and progesterone levels were measured from serum and follicular fluid. Leptin concentrations were found to be significantly higher in luteal phase follicular fluid of small follicles (P < 0.05). These were classified as atretic follicles. There was a positive correlation between serum and follicular fluid leptin levels in the luteal phase. Serum leptin was found to have a positive correlation with follicular fluid progesterone level (P = 0.01) in the preovulatory follicles. The present study shows that there is a relationship between the concentration of leptin in follicular fluid and atresia in small follicles. PMID- 16363148 TI - Effect of prepubertal ovariohysterectomy on bone mineral density and bone mineral content in puppies. AB - In this study, dual energy X-ray absorptiometry (DXA) was performed on the calcaneus (CAL) and accessory carpal bone (ACB) of early-age neutered (n = 11) and sham-operated puppies (n = 10) to investigate changes in bone mineral density (BMD) and bone mineral content (BMC) over time. Prepubertal ovariohysterectomy and sham surgery were performed at 10 weeks, while BMD and BMC were measured at 12, 16, 20 and 24 weeks of age. BMD and BMC of CAL and ACB of ovariohysterectomised puppies were found to be higher. The differences between the groups were statistically unimportant; however, the fourth measurement of the ACB showed a significant (P < 0.05) difference. Sexually intact puppies weighed less than gonadectomised puppies in all measurements. Because of the positive correlations between body weight and bone mineral density, BMD and BMC of early age neutered puppies were found to be higher. These results suggest that BMD and BMC are not affected by prepubertal ovariohysterectomy until six months of age in mixed-breed puppies. PMID- 16363149 TI - Selective apoptotic behaviour of bovine herpesvirus 1 in an epithelial-like microenvironment. AB - Apoptosis seems to play an important role in the pathogenic profile of bovine herpesvirus 1 (BHV-1) infection. Nitric oxide (NO) is also important as a signal molecule. In this study, apoptosis was selectively induced in HEp-2 cells in the early stage [1-3 h postinfection (PI)] of BHV-1 multiplication, and this apoptotic process was realised through the caspase-8, and partially through the caspase-3, pathway. BHV-1 infection inhibited staurosporine- (SS-) induced apoptosis only if the SS was added at 6 h PI. The results of this study showed that the 'NO-apoptosis' relation was realised through the caspase-8 pathway ('outer membrane receptor' pathway) at a later stage of infection in apoptosis induced by BHV-1 + SS. Our previous report (Yazici et al., 2004) and this study together showed that BHV-1 might induce and inhibit cell-type-specific pathways of apoptosis. PMID- 16363150 TI - Haematological and histological findings in birds experimentally infected with highly pathogenic H5N2 avian influenza virus. AB - Experimental infection of specific-pathogen-free (SPF) Leghorn chickens with a highly pathogenic H5N2 avian influenza virus produced cellular hyperplasia in the bone marrow at 36 hours post infection (hpi) and haematological evidence of monocytosis, thrombocytopenia and heterophilia was also detected. An early, significant and progressive haematological change was thrombocytopenia starting at 24 hpi without an increase of prothrombin time. The findings suggest that highly pathogenic avian influenza virus interferes only with the primary haemostatic mechanisms by consumption of thrombocytes, while the secondary haemostatic mechanisms remain intact. PMID- 16363151 TI - Haematological and histological findings in Leghorn chickens infected with infectious bursal disease virus strain 73688. AB - In the present study, specific-pathogen-free, 2-week-old Leghorn chickens were experimentally infected with strain 73688 of infectious bursal disease virus (IBDV) in order to evaluate haematological and histological changes that might suggest a pathomechanism for haemorrhages in this disease. At 96 hours post infection (hpi) a significant increase in prothrombin time was detected in the absence of visible lesions in myeloid bone marrow tissue and of significant thrombocytopenia. The aforementioned findings suggest alteration of the secondary coagulation mechanisms and not a direct effect of virus on thrombocytes or its precursors. PMID- 16363152 TI - Thrombocytopenia in Newcastle disease: haematological evaluation and histological study of bone marrow. AB - A Newcastle disease virus (NDV) isolated in Mexico and called Chimalhuacan strain was characterised by gene F restriction enzyme analysis and found to be a genotype II velogenic virus. Haematological evaluations and histological studies of bone marrow were conducted on chickens experimentally infected with the Chimalhuacan virus and on control chickens. Within 72 hours post infection (hpi), a 50% decrease in thrombocyte and monocyte counts and a complete cellular depletion in bone marrow islands were evident in the infected group. These findings suggest that the Chimalhuacan strain of NDV causes an early and severe damage of the haematopoietic cells including thrombocyte precursors, which might explain the marked thrombocytopenia detected in early stages of this disease. PMID- 16363153 TI - Eradication of Aujeszky's disease from a large-scale pig farm. AB - Eradication of Aujeszky's disease from a large-scale pig farm by the 'test-and cull' eradication procedure based on the use of gene-deleted vaccines and serological monitoring using gE ELISA is described. Aujeszky's disease free status was successfully achieved on a farrow-to-finish type farm characterised by three difficulties interfering with the success of eradication: (i) weaned (30 day-old) piglets remained in the farrowing house until 65 days of age; (ii) the farm did not use nursery rearing; (iii) fatteners were kept together with the breeding gilts. Thus, the immunisation programme had to be adjusted to the management technology used on the farm, and by regular serological monitoring it had to be periodically modified to make it suitable for attaining the goals. This meant that breeding animal candidates of a wide age range and kept in the same air space had to be provided with a continuously high-level immunity by regular vaccination sufficient even for rectifying potential human errors. The immunisation programme was designed by taking into consideration the disappearance of maternal immunity, and a strict vaccination programme was applied during the rearing of breeding gilts kept together with the fattening pigs. Frequent immunisation applied during the rearing of the own breeding animal replacements proved to be of decisive importance for the eradication of Aujeszky's disease from the entire herd. In addition to the accurate implementation of the vaccination programme, regular serological monitoring also made a major contribution to the success of eradication. PMID- 16363154 TI - Antennal phenotype of Triatoma dimidiata populations and its relationship with species of phyllosoma and protracta complexes. AB - Triatoma dimidiata (Latreille 1811) Reduviidae Triatominae is the main vector of Chagas disease in several countries of Latin America. As for other vector species, the characterization of T. dimidiata subpopulations within particular geographical regions or occupying different habitats could help in better planning of vector control actions. A first objective in this study was to evaluate the antennal phenotype as a phenetic marker to characterize populations of T. dimidiata collected in different geographic areas and domestic and sylvatic habitats. A second objective was to evaluate the phenetic relationships of T. dimidiata with other species of the phyllosoma complex: longipennis, pallidipennis, and phyllosoma. The antennal sensilla of T. dimidiata specimens collected in Mexico, Central America, and Colombia were analyzed and compared with the antennal sensilla of T. longipennis, T. pallidipennis, and T. phyllosoma. T. barberi was used as an outgroup in the analysis. For each specimen, the ventral side of the three distal segments of the antennae was drawn, identifying and counting four types of sensilla. In T. dimidiata, univariate and multivariate analysis showed differences between sexes, among populations collected in different habitats within the same region, and among populations collected in different geographic regions. Two types of antennal sensilla showed a latitudinal variation. Domestic specimens showed intermediate characteristics of the antennal phenotype, between sylvatic cave- and sylvatic forest-collected specimens. The antennal phenotypes show high similarities among T. pallidipennis, T. phyllosoma, and T. longipennis, with a better differentiation of T. pallidipennis. T. dimidiata is separated from the other members of the complex by a similar distance to T. barberi, of the protracta complex. PMID- 16363155 TI - Geographic distribution and breeding site preference of Aedes albopictus and Aedes aegypti (Diptera: culicidae) in Cameroon, Central Africa. AB - Presence in Cameroon of the recently introduced Aedes (Stegomyia) albopictus (Skuse) in association with the indigenous Aedes aegypti (L.) raises public heath concerns because it might alter the risk of arbovirus transmission. The breeding site and distribution of the two Stegomyia species are updated and reported following entomological surveys carried out in 22 localities throughout Cameroon, with a total of 1,353 containers with water visited. Ae. aegypti was found in every location sampled, showing higher infestation rates in northern Cameroon. Breeding populations of Ae. albopictus were observed in all 19 southern localities, up to the Adamaoua mountains, but the species was not recorded further north. In the area where both species are present, they were often sampled in the same larval developmental sites, suggesting convergent habitat segregation. The most frequently encountered artificial and natural breeding sites were used tires, discarded tins and plastic containers, abandoned car parts, brick holes, dead leaves on the ground, tree holes, and rock pools. Further monitoring of the demographic as well as geographic expansion of Ae. albopictus in this Afrotropical environment and its relationships with indigenous Ae. aegypti should provide insight into the biology of this highly invasive species and help to implement arboviruses surveillance programs in the area. PMID- 16363156 TI - Spread of Musca domestica (Diptera: muscidae), from two caged layer facilities to neighboring residences in rural Ohio. AB - Complaints of excessive numbers of flies were reported by citizens living in a rural area surrounding a large (>2 million laying hens) egg-layer facility in northwestern Ohio. Sticky cylinder traps and hanging fly strips were used at outdoor and indoor locations, respectively, at known distances from the layer farm and from control sites to determine the most likely source of the flies and to determine the severity of the problem compared with fly populations in nearby rural settings. House flies, Musca domestica (L.), were the predominant flies captured on fly traps located within 6.4 km of the poultry operations. There was a significantly greater number of M. domestica trapped in the study area surrounding the layer facility than in the control areas. The quantity of house flies captured decreased with increased distance from the layer farm. Two years into the study, a second egg-layer facility opened in an area that was originally a control site. With regard to this second farm, after 4 yr of study, there was a significant difference shown between the population of house flies during the 2 yr control phase and the 2-yr period when the egg-layer facility was operational. This study documented that large egg layer facilities can significantly increase the house fly population in the surrounding community up to 6.4 km from the source of the flies and may result in a severe nuisance up to 3.2 km away. PMID- 16363157 TI - Aspects of bioecology of two Rift Valley Fever Virus vectors in Senegal (West Africa): Aedes vexans and Culex poicilipes (Diptera: Culicidae). AB - The dispersal, population dynamics, and age structure of two Rift Valley Fever Virus (Phlebovirus: Bunyaviridae) (RVFV) vectors, Aedes vexans Meigen and Culex poicilipes Theobald, were investigated in northern Senegal. The main objective was to investigate possible factors that mediate RVFV emergence and propagation at a site where humans and livestock live in proximity to temporary surface pools. In mark-release-recapture studies, recapture rates of 0.18% (156/85,500) and 3.46% (201/5,800) were obtained for Ae. vexans and Cx. poicilipes, respectively. The number of mosquitoes recaptured decreased with increasing distance from the release point and over time. The estimated daily survival rate for released females ranged from 91 to 96% for Ae. vexans and 70-79% for Cx. poicilipes. The maximum time after release when marked mosquitoes were collected was 23 and 12 d for Ae. vexans and Cx. poicilipes, respectively. The maximum distances from the release point that marked females were recaptured was 620 and 550 m for Ae vexans and Cx. poicilipes, respectively. Rainfall periodicity was a key factor controlling Ae. vexans population abundance. In contrast, rainfall had no discernible effect on the fluctuation of Cx. poicilipes numbers. The involvement of these two species in the transmission of RVFV is discussed with respect to their longevity and daily survival rate. PMID- 16363158 TI - Microhabitats occupied by Loxosceles intermedia and Loxosceles laeta (Araneae: Sicariidae) in Curitiba, Parana, Brazil. AB - A survey was done of the environments and substrata occupied by Loxosceles intermedia Mello-Leitao, 1934 and Loxosceles laeta (Nicolet, 1849) (Araneae: Sicariidae) in urban woods and anthropic constructions in Curitiba. In total, 2,099 trees, 364 decaying logs as well as stones and roots, built-up areas, and wasteland in six urban parks and 60 residences were inspected. In total, 1,775 m2 of vegetation was inspected, but spiders and their vestiges were collected only in and around buildings in urban parks and residences. L. intermedia was more common than L. laeta and occurred both indoors and outdoors, whereas L. laeta was more common indoors in wooden houses. The two species did not occur in the same microhabitats, although both preferred paper, wood, and construction materials. Spiders collected in urban parks were heavier than those collected in residences, although only males collected in urban parks were larger than those from residences. The lack of vestiges indicative of a previous occupation in the urban parks suggested that both species occupied primarily the anthropic environment where they found numerous substrata that offered thermal isolation and suitable conditions for web fixation, ecdysis, and reproduction. PMID- 16363159 TI - Effects of inorganic nitrogen enrichment on mosquitoes (Diptera: Culicidae) and the associated aquatic community in constructed treatment wetlands. AB - Ammonium nitrogen (NH4-N) is a significant component of municipal and agricultural wastewaters, and nitrogen reduction is an important use of constructed treatment wetlands. The effects of ammonium nitrogen enrichment on resources of larval mosquitoes, larval mosquito abundance, adult mosquito production, and the abundance of related wetland organisms were examined in 0.1 ha replicate treatment wetlands. The hypothesis of a bottom-up effect induced by ammonium addition was not supported by bacterial abundance, mean bacterial cell size, or algal biomass in the water column. There was, however, a significant negative correlation between bacterial cell length and Culex tarsalis Coquillett (Diptera: Culicidae) larval abundance 1 wk later in wetlands enriched with ammonium nitrogen. Larval mosquito (Culex spp.) abundance in southern California wetlands enriched with NH4-N (mean approximately equal to 3 mg/liter) was significantly greater than in control wetlands at ambient nitrogen levels (8.3 mg NO3-N/liter, 0.1 mg NH4-N/liter). Adult mosquito production was nine-fold greater and chironomid larvae were significantly more abundant in wetlands enriched with NH4-N than in controls but other censused taxa exhibited no significant trends. Mosquitofish, Gambusia affinis (Baird & Girard), abundance was significantly reduced in enriched wetlands, but other potential mosquito predators were not significantly affected by ammonium enrichment. PMID- 16363160 TI - Asymptomatic carriers of Plasmodium spp. as infection source for malaria vector mosquitoes in the Brazilian Amazon. AB - We have described the existence of asymptomatic carriers of Plasmodium vivax and Plasmodium falciparum infections in native Amazon populations. Most of them had low parasitemias, detected only by polymerase chain reaction (PCR). Because they remain symptomless and untreated, we wanted to determine whether they could infect Anopheles darlingi Root, the main Brazilian vector, and act as disease reservoirs. Fifteen adult asymptomatic patients (PCR positive only) were selected, and experimental infections of mosquitoes were performed by direct feeding and by a membrane-feeding system. Seventeen adult symptomatic patients with high parasitemias were used as controls. We found an infection rate in An. darlingi of 1.2% for the asymptomatic carriers and 22% for the symptomatic carriers. Although the asymptomatic group infected mosquitoes at a much lower rate, these patients remain infective longer than treated, symptomatic patients. Also, the prevalence of asymptomatic infections is 4 to 5 times higher than symptomatic infections among natives. These results have implications for the malaria control program in Brazil, which focuses essentially on the treatment of symptomatic patients. PMID- 16363161 TI - Cuticular hydrocarbon variability among Triatoma dimidiata (Hemiptera: Reduviidae) populations from Mexico and Guatemala. AB - The geographic variation in the cuticular hydrocarbon pattern among 11 populations of Triatoma dimidiata Latreille (Hemiptera: Reduviidae: Triatominae) from different regions of Mexico and Guatemala, was studied using capillary gas chromatography. T. dimidiata populations were differentiated based on the relative amounts of 71 hydrocarbon components. Insect population classification was mostly in agreement with their geographical vicinity; Mexican populations from the Yucatan peninsula grouped together with those from northern Guatemala, insects from the Mexican Gulf coast states were closely related to those collected from northern Oaxaca, and to a lesser extent, to insects from Chiapas. Insects from southern Oaxaca were clustered together with those from southern Guatemala. All these populations were clearly separated from Guatemalan specimens collected in caves from Alta Verapaz. PMID- 16363162 TI - Light deprivation affects larval development and arrestin gene expression in Anopheles stephensi. AB - The role of light exposure on the final stages of development of Anopheles stephensi larvae to pupae and adult mosquitoes was explored. We demonstrated a significant reduction in the development of adult mosquitoes when larvae were bred in the absence of light compared with the control group bred in alternating 12 h of light and 12 h of dark. To correlate these findings at the molecular level, RNA levels of the visual arrestin gene were examined. Arrestins are an important gene family that play a role in the vectorial capacity of Anopheles and mediate neurotransmission as well as olfactory and visual sensory reception in insects. Semiquantitative polymerase chain reaction showed a reduction in the expression of the visual arrestin gene in pupae that developed from larvae in the absence of light compared with larvae bred under normal conditions. PMID- 16363163 TI - Chromosomal localization of two antihemostatic salivary factors in Simulium vittatum (Diptera: Simuliidae). AB - The chromosomal locations of two genes encoding the salivary protein products Simulidin (SVAT) and Simulium vittatum erythyma protein (SVEP) were identified using high-resolution cytogenetic mapping. Chromosomal subsection levels were determined from larval and adult salivary gland polytene chromosomes and adult Malphigian tubule chromosomes. Syntenic relationships occurred for both loci from a wild population of Simulium vittatum IIIL-1 collected in central Alabama and the colonized IS-7 S. vittatum sibling. cSVAT mapped to the short arm of chromosomes III, IIIS-72a4.5 and cSVEP mapped to the long arm of chromosome III, IIIL-96b1. cSVAT sits proximal to a common IIIS-2 paracentric inversion, which occurs predominately as the standard sequence in the IIIL-1 sibling and as the inverted sequence in the IS-7 sibling. cSVEP sits close to the differentiated X chromosomes in the IIIL-1 sibling. PMID- 16363164 TI - Susceptibility of Musca domestica and Chrysomya megacephala to Permethrin and deltamethrin in Thailand. AB - Susceptibility to permethrin and deltamethrin was assessed in the house fly, Musca domestica L., and blow fly Chrysomya megacephala (F.). The laboratory strain of M. domestica was captured in Muang District (downtown), Chiang Mai, northern Thailand, and the field strains were collected from Muang District, two agricultural areas of Hang Dong District and San Khampaeng District. These strains displayed susceptibility to both permethrin and deltamethrin insecticides, with LD50 values of 0.0049, 0.0019, 0.0003, and 0.0005 microg/fly and 0.1058, 0.0185, 0.1251, and 0.0981 microg/fly, respectively. Tests on the laboratory strain of C. megacephala and the field strains collected from four areas of Chiang Mai (Muang District [downtown], two agricultural areas of Hang Dong District, and San Khampaeng District) and the wooded area near the National Forest revealed slightly lower susceptibility compared with house fly, with LD50 values of permethrin equal to 0.0028,0.0027,0.0024, 0.0007, and 0.0034 microg/fly and of deltamethrin equal to 0.0461, 0.0267,0.1327,0.1151, and 0.0244 microg/fly. These results indicated that the adult populations of both fly species were susceptible to both insecticides. PMID- 16363165 TI - Chlorpyrifos resistance in mosquito Culex quinquefasciatus. AB - Two mosquito strains of Culex quinquefasciatus Say, MAmCq and HAmCq, were collected from Mobile and Huntsville, AL, respectively, after the control of mosquitoes with insecticides proved difficult. A synergism study showed that resistance to chlorpyrifos in MAmCq and HAmCq was not suppressed by piperonyl butoxide (PBO) and S,S,S,-tributylphosphorotrithioate (DEF), suggesting that P450 monooxygenase- and hydrolase-mediated detoxication does not contribute to chlorpyrifos resistance in either strain. Diethyl maleate (DEM) did not cause any significant change in the level of chlorpyrifos toxicity to HAmCq. However, DEM enhanced toxicity of chlorpyrifos to MAmCq 2.5-fold, indicating that glutathione S-transferase (GST)-mediated detoxication may play a minor role in the resistance of MAmCq. An inhibition study of acetylcholinesterase (AChE) by chlorpyrifos showed that bimolecular rate constants (Ki) of chlorpyrifos for the inhibition of AChE in adults and larvae of the susceptible S-Lab strain were 2.2- and 1.9-fold higher, respectively, than in the HAmCq strain and 3.4- and 3.8-fold higher than in the MAmCq strain. The single mutation, G119S, resulting from a single nucleotide polymorphism (SNP), G to A, in ace-1 acetylcholinesterase gene was present in HAmCq and MAmCq mosquitoes. The frequency of the heterozygote for the G119S mutant allele in the HAmCq and MAmCq mosquito populations was 0.25 and 0.45, respectively, and no individuals in either of these mosquito strains were homozygous for the A allele. It thus seems likely that the presence of heterozygous individuals for the G119S allele in HAmCq and MAmCq populations may be a response to the insensitivity of AChE observed in these two mosquito strains. PMID- 16363166 TI - Insecticide susceptibility of Aedes aegypti and Aedes albopictus across Thailand. AB - Aedes aegypti (L.) and Aedes albopictus (Skuse), two important vectors of dengue fever and dengue hemorrhagic fever, were collected from Mae Sot, Nakhon Sawan, Nakhon Ratchasima, Surat Thani, and Phatthalung, Thailand, from July 2003 to April 2004. The patterns of insecticide susceptibility to temephos, malathion, and permethrin of both Ae. aegypti and Ae. albopictus larvae were determined. Ae. aegypti from all study sites were resistant to permethrin, they but were susceptible to malathion. Resistance to temephos was detected in all strains of Ae. aegypti, except those from Nakhon Ratchasima. Ae. albopictus larvae had low levels of resistance to all three insecticides, except Mae Sot and Phatthalung strains, which were resistant to permethrin. PMID- 16363167 TI - Identification of sodium channel mutations in human head louse (Anoplura: Pediculidae) from Denmark. AB - The presence of the two mutations T9291-L932F in the voltage-sensitive sodium channel gene associated with permethrin resistance (kdr-like) was shown in head louse, Pediculus humanus capitis De Geer, populations in Denmark. The existence of one susceptible and one T929I-L932F haplotype based on the limited single nucleotide polymorphism (SNP) of these sequences was established. One louse had an SNP causing a G943A substitution in the trans-membrane segment five of domain II on the sodium channel, which has not been identified in other insect species. A polymerase chain reaction-restriction endonuclease method using genomic DNA to discriminate between resistant and susceptible alleles in head lice was developed and implemented. The SNP that results in the T9291 substitution also creates a cutting site for the restriction endonuclease SspI, and the presence of one or two SspI cutting sites in head lice is diagnostic for the T9291-L932F haplotype. PMID- 16363168 TI - Effectiveness of methoprene, an insect growth regulator, against temephos resistant Aedes aegypti populations from different Brazilian localities, under laboratory conditions. AB - The susceptibility of Aedes aegypti (L.) larvae from several Brazilian populations to the juvenile hormone analog methoprene and the organophosphate insecticide temephos were investigated. Populations from Natal (northeastern region), Macapa (northern region), and Jardim America, Rio de Janeiro (southeastern region) are temephos-resistant (RR90 = 24.4, 13.3, and 15.8, respectively), whereas populations from Presidente Prudente (southeastern region) and Porto Velho (northern region) exhibit only an incipient temephos-altered susceptibility status (RR90 = 1.8 and 2.6, respectively). Biochemical assays revealed alterations of the enzymes implicated in metabolic resistance, glutathione S-transferase, mixed function oxidases and esterases, among these populations. Dose-response assays showed at most a low resistance to methoprene of all populations tested, irrespective of their temephos resistance level. However, sequential exposure of Macapa and Natal populations to temephos and methoprene indicated a potential cross-resistance when larvae are exposed to both insecticides. Nevertheless, susceptibility of the Brazilian Ae. aegypti populations to methoprene alone suggests this insect growth regulator could substitute for temephos in the control of the dengue vector in the country. PMID- 16363169 TI - Culex restuans (Diptera: Culicidae) relative abundance and vector competence for West Nile Virus. AB - The abundance and vector competence of Culex restuans Theobald and Culex pipiens L. were compared to determine the relative importance of these species as West Nile virus (WNV) vectors in the northeastern United States. Abundance was estimated from egg raft collections at 12 sites in Albany, Suffolk, and Richmond counties, New York, during July, August, and September 2002 and 2003. Cx. restuans was more abundant than Cx. pipiens in both urban and rural areas, comprising 86% of 1,623 egg rafts collected. Vector competence for WNV was estimated after feeding on an artificial bloodmeal and in vitro transmission assays. The vector competence of the two species for WNV was similar, but the dynamics of infection seems to be mosquito species dependent. These findings suggest an important role for Cx. restuans in WNV transmission cycles in New York. PMID- 16363170 TI - Blood feeding patterns of Aedes aegypti and Aedes albopictus in Thailand. AB - Aedes aegypti (L.) and Aedes albopictus (Skuse) were collected with aspirators from Mae Sot, Nakhon Sawan, Nakhon Ratchasima, Surat Thani, and Phatthalung study sites in Thailand from July 2003 though April 2004. The sandwich-B enzyme-linked immunosorbent assay was used to analyze 1,021 blood-fed specimens. Ae. aegypti almost exclusively fed on humans (99%, 658/664) in single host species, and 97% (86/88) of multiple-host bloodmeals included at least one human host. A low frequency of other hosts, including bovine, swine, cat, rat, and chicken were detected, but they represented <1% of bloodmeals. An even higher percentage of human feeding was detected in Ae. albopictus. Hosts of Ae. albopictus collected from sites in southern Thailand were entirely human (100%, n = 105) from both single and mixed meals. In the small number of double-host meals from Ae. albopictus, we detected 3.8% as swine-human and <1% from dog-human and cat-human. Forage ratios for Ae. aegypti indicated that human, dog, and swine were preferred hosts in order of preference. In contrast, bovine and chicken were avoided hosts for this species in Thailand. PMID- 16363171 TI - Borrelia burgdorferi sensu stricto in yellow-necked mice and feeding Ixodes ricinus ticks in a forest habitat of west central Poland. AB - Wild rodents and the subadult Ixodes ricinus (L.) ticks infesting them were examined for the presence of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner s.l. in a sylvatic habitat in west central Poland during May-September 2002. In total, 818 feeding ticks were recovered from 73 infested yellow-necked mice, Apodemus flavicollis Melchior; in addition, bank voles, Clethrionomys glareolus Schreber, were rarely captured and proved to be weakly parasitized. Only 2.7% of A. flavicollis and 2.2% of 320 engorging larvae were polymerase chain reaction (PCR) positive for the bacterium. All spirochete-PCR positive samples yielded exclusively B. burgdorferi s.s. This genospecies was also the most prevalent in questing nymphs and accounted for 87.5% of the total number of Borrelia infections in nymphal ticks collected during May and June 2 yr later. The presence of the same genospecies both in naturally engorged larvae and blood-positive animals as well as the high predominance of B. burgdorferi s.s. in questing nymphs strongly differs from most study sites investigated in Europe. This unique pattern of Borrelia-diversity in both rodents and ticks seems to be determined by highly site-specific host vertebrate cenosis, and yellow-necked mice are involved in the maintenance of B. burgdorferi s.s. in the forest habitat. However, the transmission efficiency of this spirochete from the mice to the I. ricinus vector seems to be very low. The research provides additional information on the complexity of B. burgdorferi s.l. ecology in Europe, pointing to the importance of the local host community. PMID- 16363172 TI - Isolations of Potosi virus from mosquitoes (Diptera: Culicidae) collected in Connecticut. AB - Potosi virus (POTV) (Bunyaviridae: Orthobunyavirus) was first isolated from Aedes albopictus (Skuse) collected in Potosi, MO, in 1989, and subsequent isolations were reported from Illinois, Michigan, Ohio, and the Carolinas. To determine whether the distribution of this virus extends into the northeastern United States, we analyzed arboviruses acquired from mosquitoes collected in Connecticut from 1998 to 2004. In 2001, a bunyavirus was isolated from Aedes vexans (Meigen) that was different from other arboviruses known to occur in Connecticut by cross neutralization and reverse transcription-polymerase chain reaction (RT-PCR) assays. Nucleotide and encoded amino acid sequences of a portion of the G2 envelope gene were 99 and 100% similar to POTV, respectively, yet distinct from indigenous strains of Jamestown Canyon (JCV), Cache Valley (CVV), and Trivittatus virus (TVTV). Viral isolates obtained from the statewide surveillance program were retested by RT-PCR coupled with restriction enzyme analysis to distinguish POTV from other bunyaviruses. POTV isolates, previously typed by neutralization, were correctly identified by RT-PCR; however, many isolates classified as JCV or CVV by enzyme-linked immunosorbent assay proved to be POTV by molecular assays. In total, 92 strains of POTV were isolated from 12 mosquito species in 2000, 2001, and 2003, whereas POTV was not detected in mosquitoes sampled during 1998, 1999, 2002, and 2004. Viral isolation rates were highest for Anopheles punctipennis (Say) (3.2-11.3 infection rate per 1,000 mosquitoes), whereas the greatest number of isolates came from Ochlerotatus trivittatus (Coquillett) (8-16 isolates). This finding represents the first detection of POTV in the northeastern United States where it infects a diverse array of mosquito species. PMID- 16363173 TI - Analysis of Barmah Forest virus disease activity in Queensland, Australia, 1993 2003: identification of a large, isolated outbreak of disease. AB - Barmah Forest virus (BFV) disease is the second most common mosquito-borne disease in Australia. Although the majority of notifications are received from Queensland, little is known about the distribution of the disease within the state, or the important mosquito vectors and nonhuman vertebrate hosts. We conducted a retrospective statistical analysis of the notifications received from Queensland residents from 1993 to 2003 to establish long-term local incidence rates and to identify disease outbreaks. In total, 4,544 notifications were received over the 10-yr period. Disease reporting peaked in autumn, although the peak transmission season encompassed both summer and autumn. Long-term standardized incidence rates for summer/autumn and winter/spring varied across the state, showing positive spatial autocorrelation in both 6-mo periods. Although 15 instances of increased disease activity were identified, only one major disease outbreak affecting eight contiguous local government areas was detected in summer/autumn 2002/2003. This outbreak contained 297 cases, 115 more than would be expected over this period. The factors important to this outbreak are unknown and require further investigation. Although the incidence rates for BFV disease are lower than Ross River virus disease, the most reported mosquito borne disease in Australia, several factors indicate that this virus should be considered an important public health risk in Queensland. These include consistent endemic transmission, apparent underreporting of the disease, and the potential for outbreaks in major population centers. PMID- 16363174 TI - Epidemic and spatial dynamics of Lyme disease in New york State, 1990-2000. AB - Lyme disease (LD) occurrence in New York State (NYS) has not only increased over time but also spread throughout the state from the original disease focus in southeastern NYS. Few studies have investigated this epidemic and spatial dynamic in great detail. Using data from the NYS Department of Health Lyme Registry Surveillance System, we summarized epidemic and spatial characteristics of LD in NYS for the 11-yr time period from 1990 through 2000. New epidemiological trends associated with age, sex, and residential influences on LD over time were found. An empirical Bayes approach was used to produce maps of smoothed incidence at different time points to give a foundation for future state and local health funding plans and education programs. PMID- 16363175 TI - Vertical and venereal transmission of Chandipura virus (Rhabdoviridae) by Aedes aegypti (Diptera: Culicidae). AB - Experiments in the laboratory documented vertical and venereal transmission of Chandipura virus (CHPV) in Aedes aegypti (L.). The minimum filial infection rate among the progeny of infected females was 1.2%; the rate among male and female progeny was 0.9 and 1.4%, respectively. The venereal infection rate of CHPV among inseminated females was 32.7%. Our study indicates the possible occurrence of vertical and venereal transmission of CHPV in insect vectors. PMID- 16363176 TI - First report of organophosphate-resistant Boophilus microplus (Acari: Ixodidae) within the United States. AB - Boophilus microplus (Canestrini), collected from Starr County, Texas, were determined to be resistant to the organophosphorus acaricides coumaphos and diazinon. Initial bioassay results from wild-collected ticks produced a probit regression slope (SE) of 3.96 (0.22), which was different from that obtained from a susceptible reference population 6.97 (0.38). Resistance ratios (RRs) (95% CI) indicated that the population was resistant to coumaphos 3.6 (3.4-3.8), 5.0 (4.5 5.5), and 6.5 (5.4-7.7) at the LC50,90,99, respectively. A second collection of wild ticks made 12 d after all cattle in the infested pasture were treated with coumaphos produced a slope (SE) that was not significantly different from a susceptible laboratory reference population. A second bioassay found these ticks to be resistant to diazinon,RR (95%CI) = 7.1 (6.5-7.7),11.7 (10.3-13.3),17.7 (14.5-21.5) at the LC50,90,99, respectively. The slope (SE) generated from the diazinon bioassay with the resistant ticks was different than that of a reference strain, 2.98 (0.12) and 6.09 (0.35), respectively. The high-dose strategy used by the Cattle Fever Tick Eradication Program was able to eradicate coumpahos resistant B. microplus after just two treatments of coumaphos, 12 d apart. PMID- 16363177 TI - Evidence of a spotted fever-like rickettsia and a potential new vector from northeastern Australia. AB - A spotted fever-like rickettsia was identified in a Hemaphysalis tick by polymerase chain reaction (PCR) amplification and sequencing of the 16S rDNA, ompA, and ompB genes. A comparison of these nucleotide sequences with those of other spotted fever group (SFG) rickettsiae revealed that the Hemaphysalis tick rickettsia ia was distinct from other previously reported strains. Phylogenetic analysis based on both ompA and ompB also indicates that the strain's closest relatives are the agents of Thai tick typhus (Rickettsia honei strain TT-118) and Flinders Island spotted fever (R. honei). This study represents the first report of an R. honei-like agent from a Hemaphysalis tick in Australia and of a spotted fever group rickettsia from Cape York Peninsula, Queensland. PMID- 16363178 TI - Embracing our ethos, reclaiming our heart. AB - A profession's ethos is an interlacing of sentiment, value, and thought that describes its character, conveys its genius, and manifests its spirit. The ethos endures over time, serving as touchstone against which individuals may strike their actions to know their worth. As a profession's inner voice, the ethos inspires individuals and calls them back if they stray too far. This discussion of the occupational therapy profession's ethos has two discernable parts. The first invites deliberation, given its historical ground. The second is a blend of ideas and images, a clutch of reflections inviting affect and will to join in thought. My hope is to illuminate the ethos of the profession in such a way that a new perspective on current challenges is possible. I set before you this idea: To advance into the future with the ethos that has characterized occupational therapy since its inception is a reclamation of the profession's heart. PMID- 16363179 TI - Developing internet-based occupational therapy services. PMID- 16363180 TI - Editor's note--question or settle? PMID- 16363181 TI - Passion and perseverance. PMID- 16363182 TI - Role competencies for a faculty member in an occupational therapy assistant academic setting. PMID- 16363183 TI - Role competencies for a program director in an occupational therapy assistant academic setting. PMID- 16363184 TI - Occupational Therapy Code of Ethics (2005). PMID- 16363185 TI - Enforcement procedures for the Occupational Therapy Code of Ethics. PMID- 16363186 TI - Complementary and alternative medicine (CAM) position paper. AB - Occupational therapy professionals facilitate proficient and satisfying engagement in the significant tasks and meaningful activities of life. Complementary and alternative medical practices, systems, and products may be appropriately incorporated into occupational therapy practice as a way to encourage a client's engagement in meaningful occupations. Scientific studies are needed to validate the safety and efficacy of CAM methods within occupational therapy practice. Advanced-level training and continuing education are important in order to acquire the knowledge and skill to utilize CAM methods, to address the concerns for patient safety and informed consent, and to meet the rigors of regulatory requirements. PMID- 16363187 TI - Telerehabilitation position paper. AB - Though the use of telerehabilitation technologies is expanding quickly as a viable method of service delivery for many practitioners within the field of health care, there remain issues of efficacy, cost, reimbursement, legal and ethical ramifications, and practitioner competence. There is a significant need for occupational therapy practitioners to document, research, and publish on the efficacy of consultation, intervention, and follow-up services provided using telerehabilitation technologies. Further investigation of the use of telehealth technologies in professional development and supervision is needed to clarify effectiveness and efficiency, as demand for services, particularly in rural areas, threatens to exceed services available. Occupational therapy practitioners using telerehabilitation methods must adhere to the AOTA Occupational Therapy Code of Ethics (AOTA, 2000), maintain the AOTA Standards of Practice (AOTA, 2005), and comply with state regulations, ensuring both their proficiencies as practitioners and the well being of their clients. PMID- 16363188 TI - Standards for continuing competence. PMID- 16363189 TI - Standards of practice for occupational therapy. PMID- 16363190 TI - Driving and community mobility. AB - Attention to driving and community mobility is a growing area of concern due to the implications across the life span, association to occupational engagement, and relevance to other organizational entities (see Appendix B). The skills, knowledge base, and scope of practice of occupational therapy enhanced by additional training in driver rehabilitation place the profession of occupational therapy in the forefront of driving and community mobility services. The focus on injury prevention, engagement in occupation, and the intervention strategies used in driver rehabilitation and community mobility services are consistent with the Philosophical Base of Occupational Therapy (AOTA, 1995) and, therefore, warrant attention in all areas of occupational therapy practice. PMID- 16363191 TI - Occupational therapy and hospice. PMID- 16363192 TI - Occupational therapy services in facilitating work performance. PMID- 16363193 TI - The scope of occupational therapy services for individuals with autism spectrum disorders across the lifespan. PMID- 16363194 TI - Maternal mortality reduction by improving emergency obstetric care (EmOC) service. PMID- 16363195 TI - Review analysis of medullary carcinoma thyroid--15-year Indian experience. AB - To emphasize the importance of adequate primary surgery in cases of medullary carcinoma of the thyroid, 44 cases of treated medullary carcinoma of thyroid were retrospectively reviewed in Government General Hospital, Chennai between 1987 and 2002. Patients who underwent total thyroidectomy with only central compartment dissection were compared with those who had undergone total thyroidectomy with meticulous triple compartment (bilateral lateral and central groups) nodal dissection. The group of total thyroidectomy with only central compartment dissection had high rate of lymph nodal recurrence and persistent hypercalcitoninaemia when compared with the group of total thyroidectomy with meticulous triple compartment nodal dissection. (Chi square value 4.503 with p<0.05). PMID- 16363196 TI - Effect of chitosan on lipid levels when administered concurrently with atorvastatin--a placebo controlled study. AB - In a placebo controlled trialthe lipid lowering effects of chitosan, a unique dietary fibre, was assessed when given along with atorvastatin 10 mg in patients with chronic coronary heart disease. Altogether 100 patients were studied. They were randomly allocated in two groups of 50 patients each. Patients of group A received atorvastatin 10 mg before dinner plus 2 g/day chitosan in two divided doses. The groupB patients received atorvastatin 10 mg plus placebo. Patients were followed up for a period of 6 weeks. There was significant reduction in mean body weight in group A patients (3.14% versus 1,29% of body weight, p<0.05). There was also a significant rise in HDL cholesterol value (3.8% versus 1.07%, p=0.02) in group A patients. However, there was no significant reduction in the mean values of total cholesterol, LDL cholesterol and triglyceride in the two groups, although group A patients showed marginally lower values. PMID- 16363197 TI - Ovarian neoplasms in adolescence and childhood--an analysis of twenty cases. AB - Between January, 1999 and June, 2001 consecutive twenty cases of ovarian tumours in girls below 20 years of age admitted in Burdwan Medical College, Burdwan, West Bengal were analysed. Commonest presentation was lump abdomen (80%), 15% presented with acute abdomen, one patient presented with virilisation and another with precocious puberty and 85% had single ovarian involvement. Among 5 cases of torsion of ovarian tumours, one along with the twisted uterus was detected. Conservative surgery in the form of cystectomy or unilateral oophorectomy with or without salpingectomy was possible in 85% cases. Germ cell tumours (60%) took the leading place followed by epithelial (3 0%) and sex-cord stromal tumour (10%). Fifty percent neoplasms were malignant with majority (80%) being instage I. Combination chemotherapy was considered for 7 out of 10 cases. Conservative surgery in one hand and the fear of malignancy on the other are the real problems in these cases. Early detection, optimal therapy and intense follow-up are the key points of the management. PMID- 16363198 TI - A comparative study of common ear morbidity pattern among the primary school children of an urban slum of Kolkata and rural area of Hooghly. AB - A cross-sectional, clinical and epidemiological study was undertaken among 627 primary school children (rural 145, urban 482) to compare the common ear morbidity pattern between an urban slum of kolkata and a rural area of Hooghly. Middle ear pathology was found to be present in 20% and 12.6% among rural and urban students respectively. Cerumen in the external auditory canal was the commonest clinical finding in both the areas and was found to be present in 35.86% of rural and 30.70% of urban population respectively. Smoke nuisance, bathing in open ponds and overcrowding were some of the predisposing factors causing ear diseases, like chronic suppurative otitis media and serous otitis media. PMID- 16363199 TI - Sweet's syndrome preceding carcinoma of the adrenal cortex. AB - A case of Sweet's syndrome preceding carcinoma of the adrenal cortex is reported. Sweet's syndrome can be a rare premalignant manifestation of carcinoma of adrenal cortex. PMID- 16363200 TI - Benign gastric teratoma--a case report. AB - A rare case of benign gastric teratoma occuring in a 9-month child is reported with a brief review of literature. PMID- 16363201 TI - Atypical presentation of a unilateral septic thrombosis of the cavernous sinus. AB - A 7 year-old boy with unilateral septic cavernous sinus thrombosis following a pyoderma in the eyebrow is described. Problems in diagnosis and the role of magnetic resonance imaging (MRI) in the management are highlighted. Patient recovered with mild residual sequelae. The unique feature of this case was absence of chemosis and non-toxic presentation of the patient. PMID- 16363202 TI - Levofloxacin in enteric fever--a study. AB - Typhoid fever is an important cause of morbidity and mortality in patients especially in developing country. Therapy with conventional drugs is associated with increasing resistance, non-compliance to therapy and toxicity. Oral fluoroquinolones have been shown to be effective compared to parenteral broad spectrum cephalosporins in the treatment of uncomplicated typhoid. However, there is no data available regarding the use of levofloxacin in the treatment of typhoid fever in spite of the susceptibility of Salmonella species to levofloxacin. The present study was undertaken to evaluate the efficacy, safety and tolerability of oral levofloxacin 750 mg once daily in the treatment of typhoid fever. Results indicated that levofloxacin 750 mg administered orally once daily was an effective, safe, well-tolerated and cost-effective option in the treatment of typhoid fever in adult Indian males and non-pregnant females. PMID- 16363203 TI - Sulphonylureas in the management of type 2 diabetes during the fasting month of Ramadan. AB - Although a majority of Muslim patients with type 2 diabetes fast during the month of Ramadan, there are no accepted guidelines for its management during this period. The few studies on this subject suggest that there are important alterations in energy intake and physical activity, and that most patients change their pattern of drug intake. This is associated with a greater risk of hypoglycaemia and ketoacidosis. The usual pattern of eating during Ramadan, and its influence on the normal diurnal variation of blood sugar with a regular non fasting diabetic diet, suggests that anti-diabetic agents for use during this period need to be selected according to their pharmacokinetic and tablet formulation characteristics. The sulphonylureas are first line drugs in type 2 diabetes and used by a majority of patients. A comparison of the pharmacokinetics, efficacy, and safety characteristics of these agents suggests that a long-acting once daily formulation of gliclazide such as gliclazide modified release, taken in the evening, may be the sulphonylurea of choice during Ramadan. PMID- 16363204 TI - Evaluation of efficacy and safety of fixed dose combination of glimepiride 2 mg pluspioglitazone 15 mg plus metformin SR 500 mg in the management of patients with type-2 diabetes mellitus. AB - An estimated 25 million Indians currently have diabetes and the projections indicate Indians would be the largest group by the year 2025 AD. An open, phase III, multicentric study was conducted to determine the efficacy and tolerability of the triple drug combination glimepiride 2 mg plus pioglitazone hydrochloride 15 mg plus metformin SR 500 mg for 8 weeks in 101 Indian patients with type 2 diabetes mellitus. The study revealed that the triple drug combination could achieve the recommended goals, recommended by American Diabetic Association, for fasting blood glucose < or = 140 mg/dl and glycosylated haemoglobin (HbA1c) of < or = 8%. After 8 weeks, the mean fasting blood glucose (baseline 189.61) was reduced to 111.68 (41% reduction); the mean glycosylated haemoglobin (baseline 10.32) was significantly reduced to 7.54 (26% reduction). The triple drug combination significantly reduced the levels of triglyceride, low density lipoproteins and total cholesterol. These significant levels were achieved within 8 weeks and all patients tolerated the drug well with no reported case of serious adverse events including hypoglycaemia. There were also no reported drug interactions in the study. Since the decrease in HbA1c was continuous and throughout the study, a further decrease in the HbA1c levels would have been noted since the present trial was designed for a period of 8 weeks. Thus, the present study confirms the efficacy and safety of FDC of the triple drug combination in patients with type 2 diabetes. PMID- 16363205 TI - [A literature review of equine piroplasmosis after an episode of acute babesiosis in a Dutch Standardbred foal after a stay in Normandy]. AB - Piroplasmosis, a disease endemic to most tropical and subtropical areas, appears to be spreading to more temperate zones. This article gives a review of equine piroplasmosis and describes an acute case of infection with Babesia caballi in a Dutch Standard bred foal after a short stay at a stud in Normandy (France). A 3 month-old stallion foal was presented with lethargy, fever of 41 degrees C, and pale mucosal membranes. Haematology revealed a low packed cell volume (14 l/l) leucytosis (25 G/l) and a high blood urea nitrogen concentration (20.1mmol/l). Infection with B. caballi was diagnosed on the basis of Giemsa staining blood smears and was confirmed by polymerase chain reaction in combination with RLB. Treatment with imidocarb dipropionate and a blood transfusion resolved the haemolytic crisis. PMID- 16363206 TI - [Scabies in business]. PMID- 16363207 TI - [Protozoan infection causes diarrhea in calves]. AB - The role of protozoan parasites in the etiology of diarrhea in calves is highlighted with emphasis on correct diagnosis. In neonatal calves, Cryptosporidium parvum is isolated in more than 44% of the faeces of diarrhetic calves. In calves older than one month, both Eimeria bovis and E. zuernii, and Giardia duodenalis are associated with diarrhea and poor growth. Clinical diagnosis has to be confirmed by examination of host faecal material. Both for C. parvum and G. duodenalis immunological assays are available. Control measures must aim to reduce or prevent oocyst or cyst transmission, by combining management measures, desinfection and chemotherapeutic treatment. PMID- 16363208 TI - [On the way to detect functional failing of the articular cartilage in the horse]. PMID- 16363209 TI - [Particularly interesting and educational 3rd HAP congress]. PMID- 16363210 TI - [Ectoparasite symposium]. PMID- 16363211 TI - [Cooperation between the pathologist and the veterinarian is of essential importance to reach an optimal diagnosis. Interview by Dr. Marianne M. Sloet van Oldruitenborgh-Oosterbaan]. PMID- 16363212 TI - [The safety of our dog and cat feed]. PMID- 16363219 TI - [Infected foot wounds in an elderly person]. PMID- 16363220 TI - [The nursing profession in France: from medical project to the emergence of a nursing discipline project]. PMID- 16363221 TI - [In the beginning it was "taking care"]. PMID- 16363222 TI - [What is the image of nurses today?]. PMID- 16363223 TI - [The construction of the nursing care discipline in France]. PMID- 16363224 TI - [What future for the profession?]. PMID- 16363225 TI - [3/4 Preventing musculo-skeletal disorders]. PMID- 16363226 TI - [III--Relationship care at the family level. 6. Helping the family after the announcement of bad news]. PMID- 16363227 TI - The role of low molecular weight heparin to prevent miscarriage in thrombophilic women. PMID- 16363228 TI - To think out of the box, one must first get out of the test tube. PMID- 16363229 TI - Potential predictors of bleeding risk in inherited factorVII deficiency. Clinical, biological and molecular criteria. AB - Due to the wide molecular and clinical heterogeneities of inherited factor VII (FVII) deficiency, consensus guidelines for management of this coagulation disorder are not currently well established. Therefore, potential clinical, plasmatic or genetic criteria, that could be predictive for bleeding tendency in this condition, have been evaluated. Genotypic criteria including FVII genotypes and thrombophilic mutations are of particular interest to better understand some of the variations observed in bleeding phenotypes but they are still poorly informative for the management of surgery in FVII-deficient patients. Up to now, no plasma parameters have been found to be reliable predictors of bleeding risk. Nevertheless, tissue factor and platelet pathways remain to be explored. Finally, clinical history appears to be the best predictor of bleeding risk after haemostatic challenges in inherited FVII deficiencies. Furthermore, the absence of history of bleeding or mild bleeding phenotypes including menorrhagia, bruises and epistaxis (not inducing iron deficiency anaemia or requiring blood substitutive treatment) could enable minor surgery to be performed in FVII deficient patients without blood replacement therapy. PMID- 16363230 TI - Homocysteine and thrombosis: from basic science to clinical evidence. AB - Homocysteine is a sulfhydryl-containing amino acid formed during the metabolism of methionine. Rapidly accumulating evidence links elevated homocysteine levels to thrombosis via several mechanisms such as increased tissue factor expression, attenuated anticoagulant processes, enhanced platelet reactivity, increased thrombin generation, augmented factor V activity, impaired fibrinolytic potential, and vascular injury, including endothelial dysfunction. Molecular mechanisms underlying prothrombotic actions of homocysteine are incompletely understood and involve oxidative stress, DNA hypomethylation, and proinflammatory effects. Current evidence from retrospective and prospective studies supports the concept that higher total plasma homocysteine concentration is associated with increased risk of coronary artery disease, stroke, and venous thromboembolism. Hyperhomocysteinemia is currently considered a relatively weak prothrombotic factor. It is still unclear whether administration of vitamins, that reduce homocysteine levels acting as cofactors of the enzymes involved in the methionine metabolism, may decrease the risk of arterial and/or venous thromboembolic events. Ongoing clinical trials might help clarify this issue. PMID- 16363231 TI - Understanding platelets. Lessons from proteomics, genomics and promises from network analysis. AB - New large-scale analysis techniques such as bioinformatics, mass spectrometry and SAGE data analysis will allow a new framework for understanding platelets. This review analyses some important options and tasks for these tools and examines an outline of the new, refined picture of the platelet outlined by these new techniques. Looking at the platelet-specific building blocks of genome, (active) transcriptome and proteome (notably secretome and phospho-proteome), we summarize current bioinformatical and biochemical approaches, tasks as well as their limitations. Understanding the surprisingly complex platelet regarding compartmentalization, key cascades, and pathways including clinical implications will remain an exciting and hopefully fruitful challenge for the future. PMID- 16363232 TI - Circulating B-type natriuretic peptides in patients with acute coronary syndromes. Pathophysiological, prognostical and therapeutical considerations. AB - Fifty percent of patients who experience death or develop heart failure after acute coronary syndromes (ACS) have extremely elevated concentrations of plasma B type natriuretic peptides. These elevations, however, seem not to reflect permanent ventricular dysfunction or heart failure and are assumed to exist already at the onset of ischemic symptoms. The underlying mechanisms of BNP/Nt proBNP elevations in patients with ACS are still not known at present. Furthermore, the relationship of elevated BNP/Nt-proBNP with mortality but not with atherothrombotic complications of underlying disease makes it difficult to choose optimal therapeutic strategies based on plasma levels of these peptides. The remarkably high short- and long-term mortality rate associated with increases of BNP/Nt-proBNP elevations clearly show the need of further investigation to focus on this high-risk group of patients in order to clarify underlying pathomechanisms and to find optimal therapeutic approaches. PMID- 16363233 TI - Combinatorial peptides directed to inhibitory antibodies against human blood clotting factor VIII. AB - The development of antibodies against blood clotting factor VIII is a major complication affecting 20-30% of hemophilia A patients receiving replacement with FVIII concentrates. This study investigated generating peptides acting as broadly neutralizing agents to block factor VIII antibodies. These peptides were selected from dual positional scanning decapeptide libraries on cellulose membranes. From this library comprising 6.8 x 10(12) peptides we selected 468 peptides for further screening rounds. Finally we identified two decapeptides with the ability to block 8 out of 10 inhibitory antibodies from sera of patients with FVIII inhibitors demonstrated by competition assays. Sequence alignment of the peptides showed similarity with several domains in the FVIII molecule demonstrating the mimotope nature of the selected peptides. Our results show the efficiency of the combinatorial library approach and show the potential of combinatorial peptides to compete out polyclonal inhibitor IgG from a broad range of patients' sera. Combinatorial peptides could be novel and highly effective drug candidates for alternative treatment in patients with factor VIII inhibitors. PMID- 16363234 TI - Protein C Sapporo (protein C Glu 25 --> Lys): a heterozygous missense mutation in the Gla domain provides new insight into the interaction between protein C and endothelial protein C receptor. AB - Interaction of the gamma-carboxyglutamic acid (Gla) domain of protein C with endothelial protein C receptor (EPCR) is a critical step for efficient activation of protein C, though interactions by mutants in the Gla domain of protein C with EPCR have been rarely evaluated. We identified a 44-year-old Japanese woman with a history of recurrent thromboembolism as an inherited missense mutation, the first such case reported in Japan, which involved a protein C Gla 25 mutation. Total protein C antigen and Gla protein C antigen levels in the proband were normal. Protein C activity measured with an anticoagulant assay was reduced, whereas that measured with an amidolytic assay was normal. She was therefore phenotypically diagnosed as type IIb protein C deficiency. Direct sequencing of the PCR fragments revealed a heterozygous G to A transition at nucleotide position 1462 in exon 3, which predicted an amino acid substitution of Glu 25 by Lys. Her mother and one son were also heterozygous for this mutation. A molecular dynamics simulation of Gla 25-->Lys/EPCR complex in water suggested that the affinity between the molecules was decreased compared to the wild type Gla domain/EPCR complex. Since Gla 25 has been shown to play an important role in protein C function, not only in membrane phospholipid binding but also in binding to EPCR, our findings provide new insight into the mechanism by which the Glu 25- >Lys mutation induces type IIb protein C deficiency in individuals. PMID- 16363235 TI - Large deletions of the PROS1 gene in a large fraction of mutation-negative patients with protein S deficiency. AB - Protein S deficiency is an autosomal dominant disorder that results from mutations in the PROS1 gene. Conventional mutation detection techniques fail to detect a pathogenic PROS1 mutation in approximately 50% of cases. The present study investigates whether large deletions of PROS1 are found in families where mutations in the PROS1 gene have not been found despite sequencing. For this purpose,a dense set of SNP and microsatellite markers were used in segregation analysis to identify deletions. Large deletions were identified by this technique in three out of eight investigated families (38%). The deletions encompassed at least 35 kb, 437 kb and 449 kb respectively. The deletions were confirmed by quantitative PCR. Haplotype analysis showed that the three large deletions and the five other disease haplotypes were all different. All of the eight disease haplotypes co-segregated with protein S deficiency, but each of the five non deletion haplotypes were present also in normal individuals. IN CONCLUSION: Large deletions of PROS1 are relatively common in protein S deficiency patients and screening for large deletions in PROS1 mutation-negative individuals are therefore warranted. PMID- 16363236 TI - Differences in the clinically effective molar concentrations of four direct thrombin inhibitors explain their variable prothrombin time prolongation. AB - Four direct thrombin inhibitors (DTIs), lepirudin, bivalirudin, argatroban, and melagatran, differ in their ability to prolong the prothrombin time (PT). Paradoxically, the DTI in clinical use with the lowest affinity for thrombin (argatroban) causes the greatest PT prolongation. We compared the effects of these DTIs on various clotting assays and on inhibition of human and bovine factor Xa (FXa). On a mole-for-mole basis, lepirudin was most able to prolong the PT, activated partial thromboplastin time (APTT), and thrombin clotting time (TCT), whereas argatroban had the least effect. At concentrations that doubled the APTT (argatroban, 1 micromol/l; melagatran, 0.5 micromol/l; bivalirudin, 0.25 micromol/l; lepirudin, 0.06 micromol/l), the rank order for PT prolongation was: argatroban > melagatran > bivalirudin > lepirudin. Although the Ki's associated with inhibition of human FXa by melagatran (1.4 micromol/l) and argatroban (3.2 micromol/l) approach their therapeutic concentrations, inhibition of FXa did not appear to be a major contributor to PT prolongation, since argatroban also prolonged the PT of bovine plasma (despite a Ki for bovine FXa of 2,600 micromol/l). Only melagatran inhibited prothrombinase-bound FXa. We conclude that the differing effects of the DTIs on PT prolongation are primarily driven by their respective molar plasma concentrations required for clinical effect. DTIs with a relatively low affinity for thrombin require high plasma concentrations to double the APTT; these higher plasma concentrations, in turn, quench more of the thrombin generated in the PT, thereby more greatly prolonging the PT. PMID- 16363237 TI - Fibrinogen Saint-Germain II: hypofibrinogenemia due to heterozygous gamma N345S mutation. AB - We have identified a novel heterozygous fibrinogen gamma chain mutation, gammaN345S (Fibrinogen Saint-Germain II), in a subject with hypofibrinogenemia. There was no evidence by mass spectrometry of plasma fibrinogen containing the mutant chain. The hypofibrinogenemia was discovered in a 26-year-old man who experienced extensive deep venous thrombosis of the left leg associated with pulmonary embolism. Investigation of potential thromboembolic risk factors revealed heterozygosity of the factor V R506Q mutation (factor V Leiden) and heterozygosity of the prothrombin gene G20210A mutation. The hypofibrinogenemia may be contributory to the thrombophilic manifestations. PMID- 16363238 TI - D-dimer levels in combination with residual venous obstruction and the risk of recurrence after anticoagulation withdrawal for a first idiopathic deep vein thrombosis. AB - We assessed the predictive value of D-dimer levels in combination with residual venous obstruction (RVO) for recurrent venous thromboembolism (VTE) in a prospective cohort of outpatients after oral anticoagulant therapy (OAT) suspension for a first episode of idiopathic proximal deep vein thrombosis of the lower limbs during a 2-year follow-up. Patients (n=400) were enrolled on the day of OAT suspension when RVO was determined by compression ultrasonography (present in 48.6% of patients). D-dimer (cut-off value: 500 ng/mL) was measured 30+/-10 days afterwards (abnormal in 56.4% of patients). The overall recurrence rate was 16.7% (67/400; 95% confidence intervals - CI -: 13-21 %). The multivariate hazard ratio (HR) for recurrence was 3.32 (95% CI: 1.78-6.75; p<0.0001) for abnormal D dimer compared to normal D-dimer and 1.2 (95% CI:0.72-2.07; p>0.05) for RVO compared to absent RVO. The recurrence rate was 5.7% (95% CI:2-13%) and 10.4% (95% CI:6-18%), respectively, for normal D-dimer either without or with RVO, 22.9% (95% CI: 14-33%) and 25.9% (95% CI: 18-35%), respectively, for abnormal D dimer, either without or with RVO. When compared with normal D-dimer without RVO, the multivariate HR for recurrence was similar for abnormal D-dimer either with RVO (4.76 - 95% CI:1.78-12.8) or without RVO (4.3-95%:1.56-11.88). Abnormal D dimer at one month after OAT withdrawal is an independent risk factor for recurrent VTE, while RVO at the time of OAT withdrawal, either with normal or abnormal D-dimer after one month, does not influence the risk of recurrence. PMID- 16363239 TI - Plasma concentrations and importance of High Mobility Group Box protein in the prognosis of organ failure in patients with disseminated intravascular coagulation. AB - High Mobility Group Box chromosomal protein 1 (HMGB1) is a nuclear DNA-binding protein acting as a proinflammatory cytokine when released in the extracellular space from necrotic cells,activated macrophages and dendritic cells. HMGB1 acts on a specific receptor, RAGE (receptor for advanced glycation end-products), and induces prolonged inflammation, organ failure, septicaemia and death. The aim of the study was to determine the diagnostic value of plasma HMGB1 concentration and its role in the development of organ failure in patients with disseminated intravascular coagulation (DIC). Plasma HMGB-1 levels were measured in patients with suspected DIC and their relationships with DIC, organ failure and clinical outcome were determined. The study took place at the intensive care facility, Mie University School of Medicine and comprised 201 patients with suspected DIC. Plasma HMGB1 was below the detection limit in normal subjects, but moderately elevated in patients with infectious diseases (4.54 +/- 8.18 ng/ml, mean +/- SD), malignancies (2.15 +/- 5.34 ng/ml),and traumas (6.47 +/- 13.13 ng/ml). DIC was associated with significantly high plasma HMGB1 (14.05 +/- 12.56 ng/ml) in these patients. The highest HMGB1 levels were in patients with organ failure (8.29 +/- 10.99 ng/ml) and non-survivors (16.58 +/- 11.01 ng/ml). HMGB1 plasma levels correlated with the DIC score and sepsis-related organ failure assessment (SOFA) score. In conclusion, our data suggest that HMGB-1 is a potentially suitable prognostic marker of OF or DIC. PMID- 16363240 TI - Modulation of systemic hemostatic parameters by enoxaparin during gestation in women with thrombophilia and pregnancy loss. AB - Recurrent pregnancy loss (PL) is associated with maternal thrombophilia and prophylaxis with low molecular weight heparin (LMWH) can improve pregnancy outcome in this setting. The aim of this study was to investigate the modulation of systemic hemostatic parameters by enoxaparin in women with recurrent PL and to evaluate plasmatic parameters that would potentially enable monitoring LMWH prophylaxis effect during pregnancy. Study group included 87 women with thrombophilia and PL treated with enoxaparin 40 mg daily vs. 40 mg twice daily. The control group comprised 40 women with normal pregnancies. Blood samples have been collected throughout the period starting at 5-10 weeks of gestation until 6 10 weeks postpartum. The determined plasmatic markers included: anti-Xa activity, total and free tissue factor pathway inhibitor (TFPI), D-dimer, prothrombin fragment 1+2 (PT1+2), activated protein C resistance (APC-SR) and free protein S. Successful pregnancy outcome was recorded in 70 (80.5%) women treated with enoxaparin, without correlation to enoxaparin dosage. Seventeen women (19.5%) had pregnancy loss at 16+/-7 (6-32) weeks of gestation. Anti-Xa levels at 10-15 weeks of gestation were higher (0.39+/-0.38 u/ml) in the successful pregnancy outcome group compared to the abortion group (0.22+/-0.2 u/ml). Prophylactic anti-Xa activity levels (0.28+/-0.13 u/ml) were documented from 15 weeks of gestation until delivery in the successful pregnancy outcome group. Significant increase in anti-Xa, total TFPI and free TFPI levels (P<0.001) was achieved after beginning of LMWH prophylaxis in successful pregnancy outcome group but not in the abortion group. D-dimer and PT1+2 levels appeared to be significantly increased while APC SR and free protein S levels gradually decreased during pregnancy, with no difference between study groups. These results suggest that LMWH prophylaxis during pregnancy enables modulation of systemic hemostatic parameters via inhibition of factor Xa and increase in plasmatic total and free TFPI levels. PMID- 16363241 TI - Few predictors of massive deep vein thrombosis. AB - Factors that predispose to thrombus propagation from the femoropopliteal veins to the pelvic veins are poorly understood. Our goal was to determine whether there are characteristics that identify patients with massive deep vein thrombosis (DVT). We compared the 122 (2.5%) patients presenting with massive DVT (pelvic plus lower-extremity DVT) to the 4,674 (97.5%) patients with isolated lower extremity DVT from a prospective United States multicenter DVT registry. Patients with massive DVT were younger (59.4+/-18.9 years vs. 64.3+/-16.8 years; p<0.01), less likely to have hypertension (40% vs. 51%; p=0.02), and more likely to smoke (21% vs. 13%; p=0.02) and have ongoing radiation therapy (7% vs. 3%; p=0.02). The massive DVT group more commonly presented with extremity edema (80% vs. 69%; p<0.01) and erythema (21% vs. 12%; p<0.01) than the isolated lower-extremity DVT group. However, after multivariable logistic regression analysis, extremity erythema (adjusted odds ratio 1.86; 95% CI 1.13-3.04) was the only independent sequela of massive DVT and younger age (adjusted odds ratio 1.17 per decreasing decade of age; 95% confidence interval: 1.02-1.34) was the only independent predictor of massive DVT. Thrombus propagation from the femoropopliteal system cannot be reliably predicted using demographic or clinical characteristics. PMID- 16363242 TI - Venous thromboembolism and fractured neck of femur. AB - The post-operative incidence of venous thromboembolism (VTE) is high for patients undergoing hip fracture surgery. Proven prophylactic measures are available although underutilized due to concern on post-operative bleeding with use of anticoagulants. This study retrospectively reviewed the clinical incidence of VTE and utilisation of thromboprophylactic protocols over an eight year period. Demographic details, mechanism of injury, VTE risk factors, prophylactic modalities (mechanical and pharmacological), operation duration, mode of anaesthesia, hospital length of stay (LOS) and post-operative complications with particular attention to suspected deep vein thrombosis (DVT) and/or pulmonary embolism (PE) were analysed. Male to female ratio was 1:2.7 with a median age of 78 years (IQR: 70-86 years) and 83 years (IQR: 77-87 years) respectively (p<0.001). Median hospital LOS was 8 days (IQR: 5-13 days) and differed with mechanism of injury. The in-hospital incidence of VTE was 1.6% (95% CI: 1.1-2.5%) with a probably underestimated three month rate of 8.2% (95% CI: 5.3-12.4%). Non fatal PE was 0.5% (95% CI: 0.2-1.0%) in-hospital and 2.6% (95% CI: 1.2-5.5%) at three months. Fatal PE was 0.5% (95% CI: 0.2-1.0%) with a three month incidence of 0.4% (95% CI: 0.1-2.4%). The in-hospital VTE incidence was kept relatively low with use of prophylactic protocols with almost all patients receiving prophylaxis by the end of the study period. Given the five-fold out of hospital increase in incidence, consideration should be given to continue prophylaxis beyond hospital discharge in this high risk group of patients. PMID- 16363243 TI - Weaknesses in the classification criteria for antithrombotic-related major bleeding events. AB - We asked two physicians to review the medical records (electronic and paper) of 100 patients on antithrombotics. The physicians used published classification criteria to identify all of the bleeding events that the patients experienced. The goal of the review was to investigate whether the physicians would identify the same antithrombotic related major bleeding events (ARMBEs) for each patient. The correct identification and classification of multiple bleeding events is a prerequisite for studies of antithrombotic treatment practices during hospitalization that predispose patients to ARMBEs. In addition, we were interested in the reasons for disagreement between the physicians, so that we could find ways of improving their agreement. The reviewers identified 299 bleeding events for the 100 patients. They disagreed on whether 29 of the events represented an ARMBE occurring during hospitalization. With a kappa statistic of 0.49 (95% confidence interval, 0.31 to 0.66) the agreement was moderate. The reviewers most often disagreed either because they misinterpreted the data (12 events) or because the classification criteria for ARMBEs were not explicit enough (9 events). Disagreement took two main forms: either the reviewers disagreed on ARMBEs by not identifying the same bleeds (11 events) or by not applying the severity criteria appropriately (7 events). Because the main type of disagreement was not identifying the same bleeds, a study investigating the antithrombotic treatment practices that predispose patients to ARMBEs would be threatened. We therefore proposed supplementing the existing classification criteria with additional rules to avoid ambiguities in patients with multiple events. PMID- 16363244 TI - Analyses of cellular multimerin 1 receptors: in vitro evidence of binding mediated by alphaIIbbeta3 and alphavbeta3. AB - Multimerin 1 (MMRN1) is a large, soluble, polymeric, factor V binding protein and member of the EMILIN protein family. In vivo, MMRN1 is found in platelets, megakaryocytes, endothelium and extracellular matrix fibers, but not in plasma. To address the mechanism of MMRN1 binding to activated platelets and endothelial cells, we investigated the identity of the major MMRN1 receptors on these cells using wild-type and RGE-forms of recombinant MMRN1. Ligand capture, cell adhesion, ELISA and flow cytometry analyses of platelet-MMRN1 binding, indicated that MMRN1 binds to integrins alphaIIbbeta3 and alphavbeta3. Endothelial cell binding to MMRN1 was predominantly mediated by alphavbeta3 and did not require the MMRN1 RGD site or cellular activation. Like many other alphavbeta3 ligands, MMRN1 had the ability to support adhesion of additional cell types, including stimulated neutrophils. Expression studies, using a cell line capable of endothelial-like MMRN1 processing, indicated that MMRN1 adhesion to cellular receptors enhanced its extracellular matrix fiber assembly. These studies implicate integrin-mediated binding in MMRN1 attachment to cells and indicate that MMRN1 is a ligand for alphaIIbbeta3 and alphavbeta3. PMID- 16363245 TI - Biphasic effects of angiotensin II and receptor antagonism on aggregability and protein kinase C phosphorylation in human platelets. AB - In order to define the role of angiotensin II (AngII) receptor subtypes, AT1 and AT2, in platelet activation, we examined the effects of AngII and receptor antagonists on both aggregability and phosphorylation status of protein kinase C (PKC) isoforms in human platelets obtained from 56 healthy volunteers. AngII promoted both spontaneous and agonist (collagen and ADP) stimulated platelet aggregation at concentrations of 10 nM or less, but the promotion effects were lost at 100 nM. Antagonism of AT1 receptor inhibited the promotion effects of AngII at 10 nM or less. On the other hand, antagonism of AT2 receptor enhanced platelet aggregability modestly with AngII at 10 nM or less, and markedly with 100 nM AngII. Furthermore, with 10 nM AngII, phospho-PKCalpha/betaII expression in platelets was increased after collagen stimulation and was inhibited by antagonism of AT1 receptor. With 100 nM AngII, expression levels of phospho PKCalpha/ betaII remained low even after collagen stimulation but were markedly enhanced by antagonism of AT2 receptor. These findings suggest that at 10 nM or below, AngII promotes aggregability and PKC phosphorylation in human platelets through the AT1 receptor, which can be inhibited by AT1 receptor antagonists, but at higher concentrations, the promotion effects were lost through the opposing action of the AT2 receptor. The present study may provide an additional mechanism for AT1 receptor antagonism, which would provide clinical benefit to patients with stroke or cardiovascular disease accompanied by hypertension. PMID- 16363246 TI - Differential gene expression in activated monocyte-derived macrophages following binding of factor VIIa to tissue factor. AB - Factor VIIa/tissue factor (FVIIa/TF) interaction has been reported to induce intracellular signalling in cells constitutively expressing TF, independently of downstream activation of the coagulation cascade. It is unknown, however, whether binding of FVII to its cofactor TF alters the gene expression profile in cells which inducible express TF under inflammatory conditions. To address this issue, gene expression patterns in cultured LPS-stimulated monocyte-derived macrophages with or without exposure to FVIIa were compared by cDNA macro-array analysis. Of the 1176 genes examined on the array, a small set of six genes (IL-6, IL-8,TNF-a, GRO-beta alpha-thymosin, cathepsin H) were consistently up-regulated and one gene suppressed (alpha-antitrypsin) in response to FVIIa in activated monocyte-derived macrophages. Among the seven genes identified by array analysis, five genes were finally confirmed by real-time RT-PCR. Interestingly, all of these genes differentially regulated in response to FVIIa (GRO-beta, IL-6, IL-8, TNF-alpha and alpha-antitrypsin) are critical in inflammation. The changes in gene expression were reflected by corresponding changes in the protein concentrations of IL-6 and IL-8 as demonstrated by ELISA. Active site-inhibited FVIIa had no effect on gene expression indicating that FVIIa-induced gene alteration is dependent on the proteolytic activity of FVIIa. The FVIIa-induced alterations in gene expression were found to be TF-dependent but independent of downstream coagulation proteins like thrombin and FXa. In summary, this study demonstrates that binding of FVIIa to its cofactor TF enhances restricted pro-inflammatory genes in activated monocyte-derived macrophages. By up-regulation of chemokines critical for leukocyte recruitment, FVIIa/TF interaction on activated monocyte derived macrophages could be relevant to prepare monocytes/macrophages for extravasation and may represent a novel amplification loop of leukocyte recruitment. PMID- 16363247 TI - Pravastatin reduces thrombogenicity by mechanisms beyond plasma cholesterol lowering. AB - Inhibitors of 3-hydroxy-3-methylglutaryl coenzymeA reductase are widely used in the management and prevention of cardiovascular disease. In addition to its major activity, plasma lipid lowering, statins have shown a wide spectrum of additional effects that may contribute to their benefits in the prevention of cardiovascular disease. Our objective was to study whether treatment with a statin, pravastatin, could reduce thrombosis triggered by damaged vessels without changing plasma cholesterol levels. A cholesterol-clamp animal model was developed by feeding swine for 100 days on an hypercholesterolemic (HL) diet; in the last 50 days, they were randomly assigned to receive either placebo (HLC) or pravastatin (5 mg . kg(-1) . day(-1)) (HLP) in addition to the hypercholesterolemic diet. A normocholesterolemic control group (NLC) was simultaneously studied. There were no significant differences in total cholesterol, LDL and HDL plasma levels between the two groups; however, mural thrombosis triggered by both an eroded and disrupted vessel wall was significantly inhibited by pravastatin (P<0.05). Axial dependence analysis of platelet deposition revealed that pravastatin treatment reduced the increase in platelet deposition associated to the shear rate increase at the stenosis. Additionally, pravastatin treatment significantly reduced platelet membrane RhoA expression (P<0.05) and vascular wall tissue factor (TF) protein expression (P<0.05). In addition to its lipid lowering effects, pravastatin can reduce blood thrombogenicity by mechanisms independent of plasma cholesterol lowering. PMID- 16363248 TI - Local delivery of 17beta-estradiol improves reendothelialization and decreases inflammation after coronary stenting in a porcine model. AB - In the current study, we investigated the effect of local intravascular delivery of 17beta-estradiol (17beta-E) on subsequent in-stent neointimal hyperplasia. Twenty-seven stents were implanted in coronary arteries of juvenile swine. Coronary arteries were randomized to local treatment with 17beta-E or no drug therapy (control-vehicle treated). Twenty-eight days post-treatment, angiographic images revealed an improved minimal lumen diameter (2.2 +/- 0.2 vs. 1.3 +/- 0.2 mm, P < 0.005) and a reduction of late lumen loss (1.7 +/- 0.2 vs. 2.3 +/- 0.1 mm, P < 0.01) in 17beta-E-treated vessels compared to control-vehicle treated. Histological analyses showed a reduction of stenosis (51.49 +/- 6.75 vs. 70.86 +/ 6.24%, P < 0.05), mean neointimal thickness (0.51 +/- 0.07 vs. 0.83 +/- 0.14 mm, P < 0.05) and inflammation score (1.29 +/- 0.28 vs. 2.85 +/- 0.40, P < 0.05) in 17beta-E-treated arteries compared to control-vehicle treated arteries. Immunohistochemistry analyses revealed a reduction of proliferating smooth muscle cells and increased in-stent reendothelialization in 17beta-E-treated arteries. Finally, we observed a correlation between neointimal hyperplasia and inflammation score, which in turn, was inversely related to reendothelialization. Locally delivered, 17beta-E is inhibiting the inflammatory response and smooth muscle cells proliferation and improving vascular reendothelialization which together are contributing to reduce in-stent restenosis in a porcine coronary injury model. Together, these data demonstrate the potential clinical application of 17beta-estradiol to improve vascular healing and prevent in-stent restenosis. PMID- 16363249 TI - Complement C3 and C-reactive protein levels in patients with stable coronary artery disease. AB - The aim of this study was to determine whether complement C3 is an indicator of coronary artery disease (CAD). We measured plasma C3 and CRP levels in 278 patients undergoing coronary angiography for typical symptoms of CAD and 269 healthy age and sex matched controls. C3 levels were significantly higher in patients compared with controls (1.15 g/l and 0.92 g/l respectively; p<0.001). In the patient group, C3 levels correlated with BMI, fasting glucose, HbA1c, fibrinogen, CRP and HDL in both men and women. CRP levels were also higher in patients compared with controls (1.14 mg/l and 0.86 mg/l respectively; p=0.005) and correlated with markers of the metabolic syndrome. In a logistic regression model including C3, smoking, hypertension, cholesterol and diabetes, C3 was independently associated with CAD with an odds ratio of 3.20 for a 1 SD increase in C3 levels. In contrast, CRP was not independently associated with CAD in a similar regression analysis. In conclusion, both C3 and CRP plasma levels are elevated in patients with symptoms of CAD. However, C3 seems to be a better indicator of CAD than CRP in this study, suggesting that C3 could be an additional marker for risk stratification in atherosclerosis. PMID- 16363250 TI - Expressed isolated integrin beta1 subunit cytodomain induces endothelial cell death secondary to detachment. AB - Expression of isolated beta integrin cytoplasmic domains in cultured endothelial cells was reported to induce cell detachment and death. To test whether cell death was the cause or the consequence of cell detachment, we expressed isolated integrin beta1 cytoplasmic and transmembrane domains (CH1) in cultured human umbilical vein endothelial cells (HUVEC), and monitored detachment, viability, caspase activation and signaling. CH1 expression induced dose-dependent cell detachment. At 24 h over 90% of CH1-expressing HUVEC were detached but largely viable (>85%). No evidence of pro-caspase-8,-3, and PARP cleavage or suppression of phosphorylation of ERK, PKB and Ikappa-B was observed. The caspase inhibitor z VAD did not prevent cell detachment. At 48 h, however, CH1-expressing cells were over 50% dead. As a comparison trypsin-mediated detachment resulted in a time dependent cell death, paralleled by caspase-3 activation and suppression of ERK, PKB and Ikappa-B phosphoyrylation at 24 h or later after detachment. HUVEC stimulation with agents that strengthen integrin-mediated adhesion (i.e. PMA, the Src inhibitor PP2 and COMP-Ang1) did not prevent CH1-induced detachment. Expression of CH1 in rat carotid artery endothelial cells in vivo caused endothelial cell detachment and increased nuclear DNA fragmentation among detached cells. A construct lacking the integrin cytoplasmic domain (CH2) had no effect on adhesion and cell viability in vitro and in vivo. These results demonstrate that isolated beta1 cytoplasmic domain expression induces caspase independent detachment of viable endothelial cells and that death is secondary to detachment (i.e. anoikis). They also reveal an essential role for integrins in the adhesion and survival of quiescent endothelial cells in vivo. PMID- 16363251 TI - Plasma markers of angiogenesis in pregnancy induced hypertension. AB - This study tests the hypothesis that abnormalities in plasma indices of angiogenesis, such as Vascular Endothelial Growth Factor (VEGF) and angiopoietins (Ang-1, Ang-2), as well as their soluble receptors Flt-1 (sFlt-1) and Tie 2 (sTie 2) respectively, are present in women with in pregnancy-induced hypertension (PIH). We also measured platelet levels of VEGF and Ang-1 (pVEGF and pAng-1 respectively). We studied 69 consecutive women with PIH (34 without proteinuria, and 35 with proteinuria, i.e. preeclampsia) who were compared to 64 consecutive women with normotensive pregnancies and 30 normotensive non-pregnant women, in a cross-sectional study. Using ELISA, we measured levels of plasma VEGF, Ang-1 & 2, Tie-2 and sFlt-1, and also the levels of angiogenic markers within the platelet [platelet VEGF (pVEGF) and platelet Ang-1 (pAng1)] by lysing a fixed number of platelets with 0.5% tween. Results show that levels of plasma VEGF, Ang-1, Ang2, sFlt-1 and Tie-2 were significantly different between the study groups. Post hoc analyses revealed plasma Ang-1 was highest in the preeclampsia group (p<0.001), whilst Ang-2 was highest in the normotensive pregnant group (p-=0.018). Plasma Tie-2 was highest in the PIH group. VEGF levels were significantly different between the preeclampsia group and the PIH group (p<0.05). Platelet VEGF levels were higher in the non-pregnant group than in the pregnant group, but there were no significant differences in the platelet levels of Ang-1 between the different groups. Ang-2, sFlt-1 and Tie-2 were undetectable in the platelet lysate in any of the patient groups or controls. Blood pressure was a major determinant of the different angiogenic factors studied. Abnormal indices of angiogenesis are evident in PIH and preeclampsia, with higher levels of sFlt-1 and lower levels of VEGF; in PIH, increased levels of Ang-1 and Tie-2, but reduced Ang-2, are evident compared to normal pregnancy. These abnormalities may have implications for the pathogenesis of PIH and preeclampsia. PMID- 16363252 TI - Plasma markers of endothelial damage/dysfunction, inflammation and thrombogenesis in relation to TIMI risk stratification in acute coronary syndromes. AB - Risk stratification at presentation with acute coronary syndromes (ACS) on the basis of the TIMI risk score for unstable angina and non-ST-elevation myocardial infarction (UAP/NSTEMI) identifies patients at high risk of recurrent cardiac events and those who benefit from more aggressive treatment strategy. We hypothesised the following: (a) that a high TIMI risk score brings a greater degree of acute changes in endothelial damage/dysfunction (circulating endothelial cells [CECs], von Willebrand factor [vWf]), inflammation (interleukin 6, IL-6) and blood thrombogenicity (plasma tissue factor, TF); and (b) that these indices are higher in those with high TIMI risk score who experienced recurrent cardiac event at day 14 and day 30. TIMI risk scores were determined at admission and 48 hours later in 88 ACS patients (60 male, age 67+/-12 yrs) with UAP or NSTEMI. CECs, IL-6 and TF levels were measured at both time points and the acute change (delta) calculated. Patients were split into high (score > or =4) or low (<4) TIMI score groups. The composite end point of death, myocardial infarction, and refractory angina requiring revascularisation following 14 and 30 days' follow-up was ascertained. Fifty-eight patients with high TIMI risk score (mean 4.7) had significantly higher baseline and 48 h CEC, vWf, IL-6,TF and deltaTF levels, compared to low TIMI risk score (mean 2.4) patients (all p<0.05). Multivariate Cox regression analysis adjusted for clinical variables and TIMI risk score expressed as either continuous or categorical variable identified baseline CECs and deltavWf levels (both p< or =0.01) as independent predictors of subsequent cardiac events at both 14 days and 30 days. TIMI risk score for UA/NSTEMI identifies those patients with more profound vascular insult, inflammation and thrombogenicity that, in the 'high risk' patient group, predicts short-term outcomes, although vascular damage was the more sensitive predictor. These indices may further refine global risk stratification for short-term adverse cardiac events in these patients. PMID- 16363254 TI - High levels of homocysteine, lipoprotein (a) and plasminogen activator inhibitor 1 are present in patients with abdominal aortic aneurysm. AB - Over the last few years,there has been increasing interest in the investigation of the pathogenesis of AAA, and a role for some novel risk factors, in particular thrombophilic risk factors, has been suggested. The aim of this study was to evaluate a number of thrombophilic parameters in a large group of patients with AAA. In 438 patients with AAA, and in 438 healthy subjects, selected to be comparable for age and gender with patients and without instrumental evidence of AAA, a pattern of thrombophilic parameters [homocysteine (Hcy), lipoprotein (a) [Lp(a)], plasminogen activator inhibitor-1 (PAI-1), anticardiolipin antibodies (ACA), MTHFR C677T polymorphism, prothrombin gene G20210A variant and FactorV Leiden mutation] has been evaluated. A significant difference for Hcy, PAI-1 and Lp(a) plasma levels has been observed between patients and controls. After adjustment for the traditional cardiovascular risk factors, a significant increased risk of having AAA has been observed for high levels of Hcy (OR: 7.8; p<0.0001), Lp(a) (OR: 2.4; p<0.0001) and PAI-1 (OR: 3.2; p<0.0001). The association has been confirmed after exclusion of patients with other localization of atherosclerosis. Moreover, a significant association between larger abdominal aortic diameters and the number of thrombophilic parameters has been reported (r = 0.13; p = 0.005). In conclusion, a significant association between abnormal levels of some metabolic parameters related to thrombosis such as Hcy, Lp(a) and PAI-1 and AAA has been observed. PMID- 16363253 TI - Effective inhibition of experimental metastasis and prolongation of survival in mice by a potent factor Xa-specific synthetic serine protease inhibitor with weak anticoagulant activity. AB - Clinical and experimental evidence suggests that the blood coagulation system is involved in the dissemination of malignant tumors. Consequently, anticoagulant agents have been tested as metastasis suppressors in experimental models. Recently, we have found a close correlation between factor Xa (FXa)-specificity of a series of synthetic serine protease inhibitors and their anti-metastatic potential in a murine T-cell lymphoma metastasis model. Interference of such inhibitors with blood-coagulation may represent a major experimental and clinical obstacle. Here, we test anti-metastatic effects of a recently developed, highly specific 3-amidinophenylalanine-type FXa inhibitor, WX-FX4, with weaker anticoagulant activity when compared to well-established FXa inhibitors, such as DX-9065a, as measured by the activated partial thromboplastin time, prothrombin time, prothrombinase complex activity, and coagulation time. Treatment of mice with WX-FX4 (1.5 mg/kg twice daily) led to significant reduction of experimental liver metastasis of a syngeneic T-cell lymphoma in DBA/2 mice (> 90%), and of experimental lung metastasis of a human fibrosarcoma in CD1 nu/nu mice (> 60%). Due to its relatively low anticoagulant activity, daily treatment over 100 days was possible, leading to significant survival benefits without inducing bleeding anomalities. FXa-inhibitors with highly efficient anti-metastatic potential without coagulation-related side effects may represent important new tools as anticancer agents. PMID- 16363256 TI - A case of thrombosis at high altitude. PMID- 16363255 TI - Multiplex fluorescent analysis of four short tandem repeats for rapid haemophilia A molecular diagnosis. AB - Indirect molecular diagnosis of hemophiliaA (HA) is carried out by analyzing intragenic polymorphic markers described along the coagulation factorVIII (FVIII) gene. Several studies have demonstrated that the two commonly used intronic short tandem repeats (STR13 and STR22) located in the FVIII gene are highly informative for this task. Two extragenic markers closely linked to FVIII (DXS1073 and DXS1108) have also been described as valuable tools for gene tracking. The objective of the present work was to develop a rapid, single-tube automated method to simultaneously analyze these four STRs. Consistent amplification was achieved by quadruplex fluorescent PCR and the products were analyzed by capillary electrophoresis. Validation of the method included DNA analysis of 88 individuals from a control population, 45 HA patients and 32 individuals from 10 HA-affected families. Statistical study showed that the STR13, STR22 and DXS1108 loci were in significant linkage disequilibrium, whereas DXS1073 was not. Nevertheless, the combination of the four markers offered a high heterozygosity rate (>90%) that improved tracing of FVIII gene inheritance. Optimal results with application to single cells in a HA preimplantation genetic diagnosis (PGD) protocol demonstrated the sensitivity of the technique. In conclusion, the automated fluorescent method described is an extremely rapid, simple and highly informative one that is easy to standardize and allows direct comparison of results among different groups working with genetic counseling, prenatal diagnosis and PGD in HA-affected families. PMID- 16363257 TI - Unusual occurrence of left ventricular thrombus with normal left ventricular wall motion in a patient receiving tamoxifen therapy. PMID- 16363258 TI - Clinical significance of hypodysfibrinogenemia gamma 326 Cys --> Tyr? PMID- 16363259 TI - Effect of unfractionated heparin and long-term treatment with low-molecular weight heparin, bemiparin, on potassium levels. PMID- 16363260 TI - Lipoprotein complexed C-reactive protein in patients with coronary heart disease. PMID- 16363261 TI - Is elevated factor VIII a risk factor for venous thromboembolism in Canada? PMID- 16363262 TI - Rapid detection of the prothrombin C20209T transition by Light Cycler analysis. PMID- 16363263 TI - Actual improvements in the treatment of von Willebrand's disease: Wilate--double virus inactivation, high purity and convenience for patients. PMID- 16363264 TI - Management of otitis media with effusion in children. AB - Otitis media with effusion in children: B-ENT Guidelines. OME is highly prevalent among young children, with peak prevalences at around two and five years of age. Although serious complications are rare, the burden of OM is large with impaired quality of life and high direct and indirect socio-economic costs. To date, medical treatment of OME is not recommended because of the limited scientific evidence that this treatment is effective in the long term. Surgical candidacy for OME depends largely on hearing status, associated symptoms, the child's developmental risk and the anticipated chance of spontaneous resolution of the effusion. Ultimately, the recommendation for surgery must be individualized. PMID- 16363265 TI - Universal newborn hearing screening. AB - Hearing loss is one of the most common congenital anomalies, occurring in approximately 1-2 infants per 1000. Left undetected, hearing impairments in infants can negatively impact speech and language acquisition, academic achievement, social and emotional development. These negative impacts can be diminished and even eliminated through early intervention at or before 6 months of age. Reliable screening tests that minimize referral rates and maximize sensitivity and specificity are available. The goal of universal neonatal hearing screening is to maximize linguistic and communicative competence and literacy development for children who are hard of hearing or deaf. Audiologic and medical evaluations should be in progress before 3 months of age. Infants with confirmed hearing loss should receive intervention before 6 months of age from health care and education professionals with expertise in hearing loss and deafness in infants and young children. PMID- 16363266 TI - Guidelines to the management of epistaxis. AB - This article is a review of the literature on epistaxis and its treatment. Data were collected from MedLine until mid September 2005, and from others readings and books. Its first goal was to present to the Belgian ENT practitioners an overview, as complete as possible, of the modern concepts in etiologies, medical, conservative, surgical (including embolization), and adjunctive therapies of epistaxis. All these topics are discussed and commented, from a medico-surgical point of view, and also from a cost effectiveness one. PMID- 16363267 TI - Management of allergic rhinitis. AB - Due to its high and increasing prevalence, its impact on quality of life, the association with multiple comorbidities and the considerable socio-economic burden, allergic rhinitis is a major respiratory disorder and represents a global health concern. The ARIA working group has proposed a new classification for allergic rhinitis into intermittent or persistent, based on the duration of symptoms. The severity of allergic rhinitis is graded according to the impact of the disease on the quality of life. The diagnosis of allergic rhinitis involves a thorough history and clinical examination. In patients suspected of having persistent AR a complete and systematic nasal examination is an absolute requirement. Anterior rhinoscopy provides limited information. Nasal endoscopy is more useful, not to confirm AR but in particular to exclude other conditions, such as polyps, foreign bodies, tumours and septal deformations. To confirm the allergic origin of rhinitis symptoms, allergy tests must be performed. The first choice test is the skin prick test. Patients with allergic rhinitis should be evaluated for asthma and patients with asthma should be evaluated for rhinitis. A stepwise therapeutic approach is recommended based on the duration and severity of disease. The treatment of allergic rhinitis consists of allergen avoidance, pharmacotherapy and immunotherapy. PMID- 16363268 TI - Management of chronic rhinosinusitis without polyps in adults. AB - Chronic Rhinosinusitis (CRS) is a group of multifactorial diseases characterised by inflammation of the mucosa of the nose and paranasal sinuses with a history of at least 12 weeks of persistent symptoms and signs despite maximal medical therapy. IgE-mediated hypersensitivity and asthma are the two most important associated factors. Diagnosis of CRS is based upon the medical history and either an endoscopy or CT scan. Initial medical treatment usually consists of the avoidance of all the contributory factors and then nasal douching and nasal steroids. If medical treatment fails, endoscopic sinus surgery is considered. The authors review the most important factors associated with CRS, the diagnostic work-up and different treatment options. They also present a rationale for treatment. PMID- 16363269 TI - Management of nasal polyposis. AB - These guidelines are modified from the recent EAACI Position Paper. Nasal polyposis is characterized by an inflammatory process, the factors of which are summarized. Recently, Staphylococcus aureus enterotoxins have been identified to modify the disease. A classification system for polyps, grading systems and epidemiologic data are given, frequent comorbidities are discussed. The diagnostic management is based on endoscopy and CT scanning. A score of severity is proposed. The therapeutic management consists of the medical treatment options, which are given with evidence-based recommendations. Surgical treatment is indicated after failure of medical treatment and commonly performed by endoscopy. Nevertheless medical therapy must be continued after surgery to prevent recurrences. Algorithms of decision are finally proposed [corrected] PMID- 16363270 TI - Cancer of the nasal vestibule, nasal cavity and paranasal sinuses. AB - The usual clinical presentation of sinonasal tumours includes symptoms that are indistinguishable from inflammatory sinus disease, namely nasal airway obstruction, pain, and epistaxis. Abnormal V1 and/or V2 sensations are a strong indication of the possibility of tumour. Computed tomography is the most reliable and informative imaging tool for evaluating the cancers of the paranasal sinuses. Magnetic resonance imaging is essential for tumour mapping because of the excellent tissue characterisation and the possibility of differentiating between neoplasms and retained secretions. A wide variety of histologies may be encountered, although squamous cell carcinoma (SCCA) is the most common. Radiation is a common adjuvant to surgery. The response of sinonasal tract tumours to radiation therapy varies with the stage and histology of the tumour. Rehabilitation after surgical resection may be accomplished with prosthodontics or reconstructive flaps. Bony erosion of the orbital walls does not constitute an indication for orbital exenteration. Patients with tumour involvement of the skull base, either in the infratemporal fossa or at the fovea ethmoidalis and cribriform plate, should be considered for craniofacial resection. Management of these tumours requires a multimodal approach, involving surgery, radiation therapy and, increasingly in recent years, chemotherapy. Management should therefore be entrusted to multidisciplinary teams only. PMID- 16363271 TI - Smell disorders in ENT clinic. AB - Olfactory disorders may have several causes. Nasal polyposis or chronic sinusitis can result in nasal obstructions that block the access of odorants to the olfactory epithelium, and this can explain the development of olfactory disorders. On the other hand, when nasal endoscopy has revealed that the nasal cleft is free of inflammatory or tumoural disease, olfactory disorders may be explained by neuroepithelial or central nervous system disturbances. This paper will provide information about current approaches to smell disorders in otorhinolaryngology. Major causes will be reviewed as outcomes after medical or surgical treatment. An algorithm will also be given to standardise clinical investigations, including psychophysical olfactory testing, imaging and electrophysiological examinations. PMID- 16363272 TI - Management of stridor in neonates and infants. AB - Stridor is the sound caused by abnormal air passage into the lungs and can exist in different degrees and be caused by obstruction located anywhere in the extra thoracic (nose, pharynx, larynx, trachea) or intra-thoracic airway (tracheobronchial tree). Stridor may be congenital or acquired, acute, intermittent or chronic. Laryngotracheal inflammation (croup) is the most common cause of acute stridor. Laryngotracheomalacia is the most common cause of congenital, chronic stridor. Stridor is a clinical sign and not a diagnosis. The golden standard in the workup of stridor is an upper and lower airway endoscopy under general anaesthesia. Endoscopic examination under general anaesthesia requires a multidisciplinary approach and close cooperation between anaesthesiologist, paediatrician, ENT surgeon and nursing staff. Following this procedure, a place in the intensive care unit should be available for those cases presenting with stridor in which a definite diagnosis could not yet be established. Although important, pre-endoscopy assessment including history, physical examination and radiological examination, is only a guide to the type and degree of pathology found during endoscopy. About 1 out of 10 infants are found to have lesions in more than one anatomical site of the upper aerodigestive tract. PMID- 16363273 TI - Initial assessment of a neck mass in children. AB - OBJECTIVE: To propose national recommendations for the initial assessment of a mass in the neck in children. METHODS: Comprehensive review of the available literature and consensus discussion with national experts in the field. RESULTS: Consensus guidelines are proposed concerning the work up of children presenting with a mass in the neck. PMID- 16363274 TI - Initial work-up in head and neck squamous cell carcinoma. AB - OBJECTIVE: To propose national guidelines for the initial assessment of head and neck squamous cell carcinoma. METHODS: Comprehensive review of the literature and consensus discussion with national experts in the field. RESULTS: Consensus guidelines are proposed concerning the work-up of patients with a presumed diagnosis of a squamous cell carcinoma of the upper aero-digestive tract. PMID- 16363275 TI - Management of neck masses in adults. AB - Initial management of a neck mass in adults is a frequently encountered problem in ENT practice. The differential diagnosis with regards to clinical presentation, localization, imaging studies and cytology and/or histology is reviewed. An algorithm is provided to help the practioner. PMID- 16363276 TI - Current evidence and prospects for medical treatment of abdominal aortic aneurysms. AB - Abdominal aortic aneurysm (AAA) is a potentially fatal condition with risk of rupture increasing as maximum AAA diameter increases. It is agreed upon that open surgical or endovascular treatment is indicated if maximum AAA diameter exceeds 5 to 5.5 cm. Continuing aneurysmal degeneration of aortoiliac arteries accounts for significant morbidity, especially in patients undergoing endovascular AAA repair. Purpose of this review is to give an overview of the current evidence of medical treatment of AAA and describe prospects ofpotential pharmacological approaches towards prevention of aneurysmal degeneration of small AAAs and to highlight possible adjunctive medical treatment approaches after open surgical or endovascular AAA therapy. PMID- 16363277 TI - [Malignancy and thrombosis: a double-sided clinical relationship]. AB - Venous thromboembolism (VTE) is the second most common cause of mortality in cancer patients and also points towards unfavourable prognosis. In about 10% of patients with idiopathic VTE there is underlying malignancy. However, the efficacy of extensive tumour screening in those patients is not yet established. Numerous plasmatic and cellular components contribute to the phenomenon of hypercoagulability in cancer patients, including Cancer Procoagulant and activation of coagulation with high levels of coagulation factors. Cancer surgery carries an increased risk compared to non-cancer patients necessating more intensive and longer thromboprophylaxis in those patients. In medical patients active cancer is associated with increased risk for VTE. In the treatment of VTE, cancer patients have a significantly higher recurrence rate for VTE when treated with vitamin K-antagonists (VKA) compared to non-cancer patients. Compared to VKA the use of low molecular weight heparin for long-term secondary prevention is more effective than vitamin K-antagonists. Thus, there is a grade 1A recommendation to use low molecular weight heparin for cancer patients during the first 3-6 months of secondary prevention of VTE. Several studies indicate that low molecular weight heparin may also improve the prognosis of cancer patients quoad vitam, particularly for cancer patients at an earlier stage of disease; this needs to be confirmed in further studies. PMID- 16363278 TI - Pulsatility index is better than ankle-brachial doppler index for non-invasive detection of critical limb ischaemia in diabetes. AB - BACKGROUND: Diabetic polyneuropathy of the feet and legs obscures the diagnosis of critical limb ischaemia (CLI) because of lack of pain sensation. Hence, the Fontaine classification does not apply to these patients. Furthermore, many of them will exhibit medial arterial calcification, which invalidates the application of sphygmomanometry. This study was done to evaluate the pulsatility index (P1) assessed at the ankle arteries by colour Doppler ultrasonography as a non-invasive method to diagnose CLI in diabetic polyneuropathy. PATIENTS AND METHODS: 140 legs of 106 diabetic patients were studied who presented with polyneuropathy and painlessness of the feet; of these, 117 feet displayed an ulcer or gangrene. CLI was defined as the need for arterial revascularisation, as indicated by the physicians in charge on the basis of a) a foot lesion Wagner grade 1-5, and b) a positive arteriography. All patients were subjected to 4 vascular assessment techniques: digital subtraction arteriography, ankle-brachial Doppler index, systolic ankle blood pressure, and PI. RESULTS: Of the 140 legs, 61 (44%) were affected by CLI, and 76 (54%) by medial arterial calcification. A PI < 1.2 indicated CLI with a sensitivity of 0.87 and a specificity of 0.62. The sensitivity and specificity of ankle-brachial index < 0.9, and of systolic ankle pressure < 70 mm Hg to predict CLI was 0.71 and 0.42, and 0.30 and 0.89, respectively. CONCLUSIONS: The pulsatility index is a better noninvasive technique than the ankle-brachial Doppler index or the systolic ankle pressure to assess critical limb ischaemia in diabetic polyneuropathy. A pulsatility index < 1.2 at the ankle arteries is a reliable criterion for diagnosis of CLI in diabetic patients with polyneuropathy. PMID- 16363279 TI - The response of skin perfusion and of rheological and immunological variables to intravenous prostanoid administration in Raynaud's phenomenon secondary to collagenosis. AB - BACKGROUND: Prostanoids are used in the treatment of Raynaud's phenomenon and acral perfusion disorders secondary to collagenosis. In subjective terms, intravenous administration of these agents produces success in more than 50% of patients. The therapeutic outcome of clinical administration of alprostadil or iloprost may vary from individual to individual. PATIENTS AND METHODS: The following variables were analysed in a cross-over study in 27 patients with collagenosis and Raynaud's phenomenon: plasma viscosity and erythrocyte aggregation (rheological variables), partial pressure of oxygen and laser Doppler flowmetry in the finger region, and lymphocyte phenotyping and interleukin (IL) determinations (immunological variables). RESULTS: Laser Doppler flowmetry revealed significant differences between patients with secondary Raynaud's phenomenon and a control group of 25 healthy subjects. Laser Doppler readings did not change significantly as a result of the treatments. Therapy with iloprost produced a reduction in IL-1beta, L-selectin (CD 62 L) and IL-6. CONCLUSION: The change in immunological variables due to iloprost may explain the long-term effects of prostaglandins in the treatment of Raynaud's phenomenon. From our results it is not possible to infer any preference for iloprost or alprostadil. PMID- 16363280 TI - Risk factors for survival in patients with end-stage renal disease undergoing infrainguinal bypass surgery. AB - BACKGROUND: A poor longevity and high perioperative morbidity make lower extremity revascularization questionable in patients with end-stage renal disease (ESRD). Therefore, careful selection of patients for surgery is essential. Aim of this study was to assess negative predictors of survival in patients with ESRD undergoing infrainguinal bypass grafting for critical limb ischemia (CLI). PATIENTS AND METHODS: We reviewed the records of 49 consecutive patients with ESRD who underwent infrainguinal bypass grafting for limb salvage. Rates were computed with life-table analysis and compared by log-rank test. Effects of demographic and disease variables on the survival rate were evaluated by Cox proportional hazard regression model. RESULTS: Indications for surgery were rest pain in two (4.1%) and tissue loss in 4 7 patients (95.9%). Median follow up was 7.8 months (IQR, 2.43 to 16.23). Perioperative (30-day) morbidity and mortality for all patients were 6.1% and 12.2%, respectively. Primary and secondary patency at two years both were 81.4%. Cumulative survival rate at two years and four years were 24.9% and 9.3%, respectively Limb salvage rate and amputation-free survival rate at two and four years were 80.4%, 53.6%, 21.8% and 14.6%, respectively. Myocardial infarction and congestive heartfailure in the patients medical history both had an adverse effect on survival rate with a hazard ratio of 5.52 (95% CI, 1.94 to 15.69) and 3.12 (95% CI, 0.99 to 9.81), respectively. CONCLUSIONS: In the presence of myocardial infarction or congestive heart failure in the medical history survival rate is especially poor for patients with ESRD undergoing infrainguinal revascularization. Therefore, bypass surgery for CLI is hardly indicated in this group of patients. PMID- 16363281 TI - Automated software supported versus manual aorto-iliac diameter measurements in CT angiography of patients with abdominal aortic aneurysms: assessment of inter- and intraobserver variation. AB - BACKGROUND: Open surgical or endovascular abdominal aortic aneurysm (AAA) relies on precise preprocedual imaging. Purpose of this study was to assess inter- and intraobserver variation of software-supported automated and manual multi row detector CT angiography (MDCTA) in aortoiliac diameter measurements before AAA repair. PATIENTS AND METHODS: Thirty original MDCTA data sets (4 x 2mm collimation) of patients scheduled for endovascular AAA repair were studied on a dedicated software capable of creating two-dimensional reformatted planes orthogonal to the aortoiliac center-line. Measurements were performed twice with afour-week interval between readings. Data were analysed by two blinded readers at random order Two different measurement methods were performed: reader-assisted freehand wall-to-wall measurement and semi-automatic measurement. RESULTS: Aortoiliac diameters were significantly underestimated by the semi-automatic method as compared to reader-assisted measurements (p < 0.0031). Intraobserver variability of AAA diameter calculation was not significant (p > 0. 15) for reader-assisted measurements except for the diameter of the left common iliac artery in reader 2 (p = 0.0045) and it was not significant (p > 0. 14) using the semi-automatic method. Interobserver variability was not significant for AAA diameter measurements using the reader-assisted method and for proximal neck analysis with the semiautomatic method (p > 0.27). Relevant interobserver variation was observed for semi-automatic measurement of maximum AAA (p = 0.0007) and iliac artery diameters (p = 0.024). CONCLUSIONS: Dedicated MDCTA software provides a useful tool to minimize aortoiliac diameter measurement variation and to improve imaging precision before AAA repair. For reliable AAA diameter analysis the reader-assisted freehand measurement method is recommended to be applied to a set of reformatted CT data as provided by the software used in this study. PMID- 16363282 TI - Lysability of arterial thrombi assessed by magnetic resonance imaging. AB - BACKGROUND: Intravascular thrombi change in time due to retraction and organization, which is reflected in the appearance of magnetic resonance images of clots. We have hypothesized that MRI has the potential to improve patient selection for thrombolytic treatment. The aim of our study was to analyze occlusive arterial thrombi with MRI, and to correlate the MRI parameters with the therapeutic outcome in patients with occlusive atherothrombotic disease of the superficial femoral artery who were treated with catheter-directed thrombolysis by streptokinase. PATIENTS AND METHODS: We included 13 patients with subacute (2 weeks to 3 months old) occlusive arterial thrombi and 4 patients with chronic (more than 6 months old) arterial occlusions. We measured the MRI signal intensity on gradient echo images of 98 axial slices of the subacute occlusive thrombi and in 45 slices of 4 chronic thrombi. Following MRI, the patients with subacute history were treated with catheter-directed thrombolysis. RESULTS: Thrombolysis was successful in 11/13 patients. The normalized MRI signal intensity was significantly higher in the unsuccessfully treated thrombi than in the successfully treated thrombi (1.10 +/- 0.08 vs. 0.72 +/- 0.17, p < 0.003), but the subacute and chronic thrombi did not differ in signal intensity. CONCLUSIONS: High signal intensity of arterial thrombi on gradient echo MRI might predict resistance to thrombolytic therapy. PMID- 16363283 TI - Follow-up after deep venous thrombosis in azygos continuation. AB - BACKGROUND: To determine the sequelae of patients after deep venous thrombosis inpatients with azygos continuation defined as agenesis of the inferior vena cava with collateral flow. PATIENTS AND METHODS: Five patients post deep venous thrombosis in the context of azygos continuation were followed up clinically and with colour duplex ultrasonography. RESULTS: All five patients had to our knowledge after the initial deep venous thrombosis no further thromboembolic events. Three patients after isolated iliac thromboses are symptom free or nearly symptom free, two after more extended thromboses still sufferfrom venous claudication. Four patients are without anticoagulation, one patient is permanently orally anticoagulated. CONCLUSIONS: Azygos continuation may not influence the risk of recurrent venous thrombo-embolism nor the outcome of a deep venous thrombosis. Careful deep venous thrombosis prophylaxis in patients with azygos continuation may be sufficient when a risk factor is present but conclusions lack due to the small numbers of patients of enough supportive data. PMID- 16363284 TI - Ultrasonographic and FDG-PET imaging in active giant cell arteritis of the carotid arteries. AB - Inflammation of the arterial wall has been demonstrated by 18 Fluoro-Deoxyglucose PET imaging in patients with Takayasu's and temporal arteritis. We used ultrasonography and FDG-PET for structural and metabolic imaging of the carotid artery to diagnose giant cell arteritis without biopsy. In a 72 years old patient with isolated clinical and paraclinical signs of severe systemic inflammation ultrasonogaphy showed concentric hypoechogenic mural thickening of the carotidarteries and high FDG uptake in the left carotid, both axillary and subclavian arteries and the aorta. Clinical and paraclinical abnormalities showed a typical response to steroid treatment. In conclusion a combined approach using ultrasound and FDG-PET seems to be helpful in the diagnosis of GCA of large arteries particularly in patients with atypical manifestations of the disease and negative or unavailable biopsy of the temporal artery. PMID- 16363285 TI - Takayasu's arteritis and ulcerative colitis in a young non-Asian woman: a rare association. AB - The association between ulcerative colitis and Takayasu's arteritis has been well described in patients of Asian ethnicity. We present the third non-Asian case reported in the English literature, of a 37-year-old woman with a 4-year history of Takayasu's arteritis, who developed idiopathic ulcerative colitis. She was found to carry the human leucocyte antigens HLA-B52 and DR2, which have been previously noted to be associated with these inflammatory conditions, mainly in the Japanese population. Ulcerative colitis was steroid-dependent despite simultaneous administration of mesalazine. Azathioprime achieved remission of both diseases. The possible pathogenic association of the disorders is discussed. PMID- 16363286 TI - Intraaortic balloon pump counterpulsation after implantation of infrarenal and thoracoabdominal aortic protheses. AB - Highly complex vascular surgery interventions have nowadays become possible due to sophisticated operative techniques and modern intra- and postoperative anesthesiological strategies. Accordingly, the number of high risk vascular surgery interventions rises continuously and thus, the number of secondary complications after high risk interventions increases as well and requires likewise extraordinary treatment concepts. We report of a 68-year old patient who 6 months previously was operated on a ruptured abdominal aneurysm, before he was admitted to our institution for the treatment of a type IIIb (Crawford classification) thoracoabdominal aneurysm. Intraoperatively we implanted a 26 mm Dacron prosthesis which was anastomosed with the previously existing infrarenal graft. Postoperatively the patient suffered from a hemodynamically significant myocardial infarction and acute coronary catheter intervention was necessary. However, circulatory stability could not be reestablished by interventional measures and we therefore decided to implant the intraaortic balloon pump despite the presence of two synthetic aortic grafts. However, the chance of success of such a manoeuver as well as the effectiveness of intraprosthetic counterpulsation was unclear and our literature research undertaken to predict the risk of such a manouver was unsatisfactory. We therefore want to report this case and compile the literature dealing with perceptions and complications of intraaortic counterpulsation after the implantation of synthetic aortic prostheses, since such a treatment option comes to an increased clinical application in comparable constellations. PMID- 16363287 TI - An extraanatomic bypass method alternative to the obturator bypass. AB - Obturator bypass is the preferred method when infectious groin problems are challenging. However, this method can not be applied either in some special settings including the dissemination of the infection towards retroperitoneum and some technical difficulties in handling obturator bypass area (like fibrosis and inability to expose the obturator foramen). We present an alternative bypass method in two cases. PMID- 16363288 TI - A review on hepatitis C virus. PMID- 16363289 TI - Fascioliasis among local and imported sheep in Saudi Arabia: parasitological and serological diagnosis. AB - Eight-hundred-twelve local and imported sheep, slaughtered at Riyadh abattoir, were subjected for parasitological diagnosis of fascioliasis by detection of eggs in the stool and worms in the liver and for serological diagnosis by detection of circulating anti-Fasciola IgG (CAFIgG) and circulating Fasciola antigens (CFAgs) using the indirect ELISA and the double antibody sandwich ELISA respectively. Detection of eggs revealed 13.5% infection rate compared with 21.9% by detection of worms (p<0.001). Infection rate was significantly higher (p<0.001) among the imported sheep (15.1%) than among the local ones (4.96%). Positivity for CAFIgG was 64.3% among all sheep while that of CFAgs was 30.2% (p<0.001). Sheep passing Fasciola eggs showed positivity of 99.1% for CAFIgG and 87.3% for CFAgs (p<0.05). Sheep free from any parasitic eggs showed positivity of 61.2% for CAFIgG and 21.1% for CFAgs. There was a highly significant association (p<0.001) between positivity of both CAFIgG and CFAgs and positivity of eggs in the stool. PMID- 16363290 TI - Immunomodulatory properties of sand fly saliva and its role on vertebrate host. AB - The salivary glands of Phlebotomus papatasi and P. langeroni were investigated for their immunomodulatory properties on vertebrate hosts. Laboratory reared sand fly, were used for feeding on hamsters and for extraction of salivary gland. To determine the influence of the sand fly saliva homogenate (SGH) were performed ELISA using sera from injected hamsters by SGH. Frequent feeding of P. papatasi on hamsters were carried out to relate the anti-body titre with the biting rate. The mosquito Culex pipiens was allowed to feed on pre-exposed hamsters to P. papatasi bites. Antibodies correlated with the saliva of both species showed low titre and not related either to the time (after feeding and injection) or to the number of fed flies. Saliva of P. langeroni (non vector) recorded higher anti body titre than P. papatasi. The preliminary experiment of the mosquitoes' feeding (C. pipiens) on pre-exposed hamsters on its saliva showed cross reactions between biting of mosquitoes and sand flies. More investigations are needed to study the effects of sand fly saliva on human immune responses to evaluate the fly salivary proteins as vaccines for leishmaniasis and to prevent or decrease the sand flies biting or probably mosquitoes. PMID- 16363291 TI - The utility of lactoferrin in differentiating parasitic from bacterial infections. AB - Lactoferrin is an iron binding glycoprotein found in the 2ry granules of PMN. In order to determine the usefulness of such marker for neutrophilic activity in differentiating cases suffering from amoebic and bacillary dysentery, Schistosoma and bacterial UTI infections, we examined stool and urine specimens using anti lactoferrin antibodies (lactoferrin latex agglutination test: LFLA), compared with different standard gold techniques. Our results demonstrated that cases with either shigllosis or UTI revealed a high lactoferrin titer which was positively correllated with the number of PMN. In addition cases with Entamoeba histolytica or S. haematobium were characterized by relatively lower inflammatory process as expressed by mild lactoferrin titer which was also correlated with the PMN count. In addition, the findings of the present work indicated that LFLA was sensitive and specific when used alone and its sensitivity was augmented after coupling with other simple indirect methods of diagnosis. In conclusion, results described the reliability of using LFLA as a simple, rapid, sensitive method in differentiating, certain parasitic from bacterial diseases. PMID- 16363292 TI - Studies on Phyllobothrium lactuca (Cestoda: Phyllobothriidae) and Philometra salgadoi (Nematoda: Philometridae) parasitizing Boops boops from the Mediterranean Sea, Egypt. AB - A total of 30 specimens of Boops boops marine fishes were collected from Alexandria fish market. The fishes were dissected out for parasites. Phyllobothrium lactuca (Cestoda: Phyllobothriidae) was isolated from the intestine and Philometra salgadoi (Nematoda: Philimetridae) was isolated from the ovary. Both were identified using standard keys, and examined by Scan Electron Microscopy (SEM). Some morphological features reported were unique. The two parasites represent new hosts record of B. boops. PMID- 16363293 TI - Biocompatible dialysis membranes and oxidative stress in patients wih end-stage renal disease on maintenance haemodialysis. AB - Oxidative stress has been shown in (ESRD) patients specially those receiving regular haemodialysis (HD) in relation with an increased production of toxic free radicals due to membrane-induced complement leukocyte activation. An imbalance between oxidants and antioxidans has been suggested in uremic patients on HD. The respective influence of uremia and dialysis procedure has not been evaluated. Studies that have probed into the mechanism of oxygen radical production have implicated the bio-incompatibility of dialysis membranes. The effect of different dialysis membranes on lipid, lipoproteins, lipid peroxidation and total antioxidant capacity in ESRD patients on regular HD was studied. One hundred subjects were selected; 20 healthy controls, 20 chronic renal failure (CRF) patients on conservative drug management and 60 CRF patients on maintenance HD (20 dialyzed by polysulfone, 20 by hemophan and 20 by cuprophane membranes). All patients were matched for age, sex, gender and etiology of ESRD and HD patients for duration of dialysis. In addition to routine tests (Hb% and creatinine clearance in healthy control group and CRF patients on conservative management), total cholesterol, triglycerides, high density lipoprotein (HDL-C) and low density lipoprotein (LDL-C), apolipoprotein A (Apo A), apolipoprotein B (Apo B), serum malondialdehyde (MDA) and plasma total antioxidant status (TAS) were estimated. MDA was significantly higher and TAS was lower in uremic patients treated conservatively or by HD than in controls. MDA was significantly higher in HD than CRF patients on conservative management with least significant difference in HD patients treated by polysulfone followed by hemophan and then cuprophane membrane, while only cuprophane group showed lower levels of TAS compared to CRF patients on conservative management. HDL-C and Apo A was higher in polysulfone and hemophan than cuprophane group while triglyderide was lower. Polysulfone group showed lower levels of LDL-C than both cuprophane and hemophane groups thus providing less atherogenic lipid profile. There was a positive correlation between Hb% and TAS and a significant negative correlation between MDA and Hb%. There was a significant negative correlation between TAS and duration of dialysis in HD patients. In CRF patients on conservative management we obtained a significant positive correlation with TAS and a significant negative correlation with MDA. PMID- 16363294 TI - Human fascioliasis among immigrant workers in Saudi Arabia. AB - Animal fascioliasis has been reported in Saudi Arabia among imported and local sheep. The paper demonstrated the parasitological and clinical features of human fascioliasis in nine out of ten male immigrant manual workers with manifestations suggesting fascioliasis. The sedimentation and Kato-Katz techniques proved effect in diagnosing Fasciola species eggs in human stool. The common clinical features were abdominal distension, flatulence, tender right-upper quadrant and easy fatigability and the least was the tinge of jaundice. Others as right upper quadrant pains, colicky abdominal pains & vomiting, epi-gastric pain and mild fever, and tympanitic abdomen were encountered. Anaemia and eosinophlia were also encountered in the ten patients. Fascioliasis patients (nine) were successfully treated with Mirazid as two capsules (600 mg) on an empty stomach an hour before breakfast for six consecutive days. Follow-up clinically and parasitologically was available in only seven fascioliasis patients who were completely cured. Follow-up for the other two fascioliasis patients was out in hand. Other parasites recovered in the stained (eosin, iodine and Zeihl-Nelson stains) smear stool samples was Entamoeba histolytica, Giardia lamblia and Cryptosporidium parvum. Besides, three were free from intestinal protozoan. The results were discussed on the light of the other work carried out regionally. PMID- 16363295 TI - What is the best medium to identify the epidural space? PMID- 16363296 TI - [Identification of the epidural space: usual practice among Spanish anesthesiologists]. AB - OBJECTIVE: To determine what methods are habitually used by Spanish anesthesiologists to identify the epidural space. MATERIAL AND METHODS: Spanish anesthesiologists were asked to fill in an Internet questionnaire about the methods they had learned during residency training for identifying the epidural space, the method they currently use most often, and the one they consider best for residents to learn during training. RESULTS: Responses were received from 617 anesthesiologists. The techniques the respondents had learned during training were loss of resistance to air (LOR-A), 58.5%; LOR to saline (LOR-S), 31.9%; LOR and air bubble (LOR-B), 6.1%; and the hanging drop method, 2.4%. Those who had learned the LOR-A technique had changed in 14.2% of the cases, as had 28.4% of those who learned the LOR-S procedure. LOR-A is currently used by 59.2% of the respondents, LOR-S is used by 32.4%, and LOR-B by 6%. The respondents recommended that new residents physicians learn the LOR-A (48.5%) and LOR-S (37.8%) techniques. The LOR-B is recommended by 12.6% a much larger percentage of anesthesiologists than it is habitually used by. Of those who use the LORA technique habitually, 26.4% recommend that new residents use a different procedure. CONCLUSIONS: The procedure used most often by Spanish anesthesiologists to identify the epidural space is the LOR-A technique; however, over a quarter of Spanish users of LOR-A do not recommend it. PMID- 16363297 TI - [Brachial plexus block with levobupivacaine at the humeral canal: comparison of a small volume at high concentration with a large volume at low concentration]. AB - OBJECTIVE: To assess differences in the brachial plexus block in 2 groups who received the same dose of levobupivacaine: 1 group received a small volume of solution at high concentration and the other group received a large volume in solution at low concentration. MATERIAL AND METHODS: A prospective, randomized clinical trial enrolling 69 patients scheduled for wrist and/or hand surgery with a brachial plexus block with levobupivacaine in the humeral canal. Nerve stimulation was used to locate a response from the 4 terminal nerves in the brachial plexus. In the group receiving a larger volume, 10 mL of a solution of levobupivacaine at a concentration of 0.375% was used for each nerve. In the high concentration group receiving a smaller volume, levobupivacaine was used at a concentration of 0.75% in 5 mL for each nerve. Sensory latency was assessed by the pin prick technique. Motor block, the success rate (percentage), and duration of sensory and motor blockades were also evaluated. RESULTS: The full sensory block was significantly more efficacious in the large volume group than in the high concentration group (85.3% vs 51.6%, P = 0.003). A full motor block was reached in a small percentage of patients in both groups. There were no significant differences in latency or duration of block. CONCLUSIONS: The success rate was lower in the group receiving the smaller volume at a higher concentration. It is advisable to administer local anesthetics in larger volumes at lower concentrations to improve block quality. Latency and duration were similar in both groups. PMID- 16363298 TI - [Preoperative use of intravenous iron: a new transfusional therapy]. AB - INTRODUCTION AND OBJECTIVE: Improving preoperative hemoglobin levels reduces transfusion requirements. The availability in Spain of iron sucrose formulas with good safety profiles has added a new therapeutic option for treating anemia. The aim of this study was to demonstrate the usefulness of preoperative intravenous infusion of iron for improving hemoglobin level before elective major orthopedic surgery. PATIENTS AND METHODS: From the beginning of 2003 until July 2004, 27 consecutive patients scheduled to undergo major orthopedic surgery were given preoperative intravenous iron infusions because of intolerance to oral iron administration, poor intestinal absorption, chronic inflammatory anemia, or functional iron deficit. Twenty patients received preoperative intravenous epoetin alfa and iron and 7 received only intravenous iron because epoetin alfa therapy was ruled out due to cardiovascular or thromboembolic disease or because they had pure iron deficiency. Variables studied were age, sex, weight, type of surgery, iron dose, duration of treatment, reason for treatment, blood parameters, hemoglobin levels, and use of transfusions. RESULTS: The preoperative increase in hemoglobin was 1.7 g dL(-1) (range, 10.8-12.5) for patients who received only intravenous iron therapy and half of them were transfused, although inappropriately in 1 case. The increase in the intravenous epoetin alfa plus iron group was 1.7 g dL(-1) (range, 10.9-12.6) and only 25% were transfused. CONCLUSIONS: Intravenous iron therapy improved the preoperative hemoglobin levels in anemic patients scheduled for major orthopedic surgery. This is a new intravenous treatment approach to evaluate. PMID- 16363299 TI - [Anesthetic management of coagulation factor V deficiency]. AB - Factor V deficiency, or Owren's disease, is a rare inherited recessive autonomic disorder that is congenital. The bleeding in patients with this disease depends on plasma levels of factor V. Anesthetic management is similar to that used for patients with other coagulation disorders. On the one hand, it is useful to initiate infusion of plasma if purified concentrates are not available. On the other hand, it is important to monitor hemostasis carefully so that bleeding during postoperative recovery can be prevented. Plasma and antifibrinolytic drugs such as tranexamic acid should continue to be administered in the first few days after surgery. We report the case of a man with homozygotic factor V deficiency who underwent surgery for colon cancer. This article reviews the main aspects of anesthetic management of this coagulation disorder. PMID- 16363300 TI - [Anterior surgical approach to T12 and L1 in recurring vertebral hydatidosis: anesthetic management]. AB - A 47-year-old man with recurring vertebral hydatidosis was scheduled for surgical removal of cysts by an anterior approach. Anesthetic management included multimodal monitoring and prophylaxis for the most common neurological, hemodynamic, and respiratory complications, as well as for appropriate control of pain during and after surgery. The spine is a rare location for hydatid cysts. Treatment is surgical, although imidazoles are useful for prevention and protection against recurrence. The prognosis is good. PMID- 16363301 TI - [Hydroxyethyl starch 130/0.4 can be useful to prevent a hemodynamic response to a single dose of remifentanil]. AB - Intravenous propofol and remifentanil are often used in anesthesia. The combined use of these drugs tends to cause hemodynamic depression. We describe the absence of hemodynamic effects in response to infusion of propofol and remifentanil when hydroxyethyl starch (HES) 130/0.4 was also administered. During induction, because blood volume needed to be replaced, two patients aged 62 and 65 years received intravenous HES 130/0.4. They then received a single dose of 2 microg x kg(-1) of remifentanil during total intravenous anesthesia (TIVA) with propofol and remifentanil before placement of a Mayfield head holder. No changes in mean blood pressure or heart rate were observed in either patient after the remifentanil bolus when they have received HES 130/0.4 during TIVA with propofol and remifentanil HES 130/0.4 may play an active role in preventing a hemodynamic response to remifentanil bolus. This hypothesis should be tested in a randomized controlled trial. PMID- 16363302 TI - [Thoracoscopy with continuous positive airway pressure and collapse of only the operated lobe]. AB - We report a case of diagnostic thoracoscopy of a right lobe in a patient with left upper lobe atelectasis due to invasion by a bronchial tumor. We anticipated difficulty in maintaining the desired oxygenation while using the conventional strategy of collapsing the whole operated lung and ventilating only the opposite one. The solution was to apply continuous positive airway pressure to 2 right lobes while only collapsing the lobe to be biopsied. PMID- 16363303 TI - [Extubation of the difficult airway]. AB - Extubation, like intubation, is a critical moment in general anesthesia. There are no algorithms or ordered sequences of steps for extubation. Rather, the approach to take is strict observation of the patient in a setting equipped with monitors, material for managing the difficult airway, and experienced staff who should be able to establish access immediately, provide oxygen, and facilitate gas exchange, keeping the airway open and safeguarding it in case of a failed extubation attempt. This review will analyze the clinical conditions and pathophysiology associated with extubations at high risk of complications. We will describe strategies for extubating in situations in which a difficult airway is known or suspected. PMID- 16363304 TI - [Severe bradycardia and spinal anesthesia in a patient with undiagnosed sinus nodal dysfunction]. PMID- 16363305 TI - [Massive abdominal wall hematoma and hemperitoneum secondary to subcutaneous infusion of low molecular weight heparin]. PMID- 16363306 TI - [Intestinal ischemia due to closed abdominal injury: a case report]. PMID- 16363307 TI - [Intradural anesthesia for cesarean delivery of a patient with Takayasu disease]. PMID- 16363308 TI - [Antihemorrhagic prophylaxis in low risk surgery in Bernard-Soulier syndrome]. PMID- 16363309 TI - [On the first and second debuts of epidural anesthesia]. PMID- 16363310 TI - [First course in perioperative medicine at Hospital de la Princesa]. PMID- 16363311 TI - [How much longer must we put up with acute pain in our hospitals?]. PMID- 16363312 TI - Accidental dural puncture in an obstetric patient, continuous spinal analgesia for labor and postdural puncture headache. PMID- 16363313 TI - [Inflammatory vascular reaction to propofol]. PMID- 16363314 TI - Seventy-two hour gas exchange performance and hemodynamic properties of NOVALUNG iLA as a gas exchanger for arteriovenous carbon dioxide removal. AB - OBJECTIVE: Acute respiratory failure is complicated by acidosis and altered end organ perfusion. NOVA-LUNG iLA is an interventional lung assist (ILA) device for arteriovenous carbon dioxide removal (AVCO2R). The present study was conducted to evaluate the device for short-term CO2 removal performance and hemodynamic response. METHODS: Six adult sheep received cannulation of the jugular vein and carotid artery. The ILA-AVCO2R circuit was placed on the sheep for 72 hours. Hemodynamics and PaCO2 were measured; CO2 removal was calculated while varying sweep gas flow rates (Qg), device blood flow rates (Qb), and PaCO2. RESULTS: Hemodynamic variables remained normal throughout the 72 hour study. CO2 removal increased with increases in Qg or Qb. Mean CO2 removal was 119.3 ml/min for Qb 1 L/ min, Qg 5 L/min, and PaCO2 40-50 mmHg. PaCO2 was directly proportional to CO2 clearance (R=0.72, p <0.001). CONCLUSION: NOVALUNG iLA can provide near total CO2 removal with Qb 1-2 L/min, Qg 5 L/min, and minimal flow resistance (3.88+/-0.82 mmHg/L/min). PaCO2 correlates with CO2 removal and is dependent on Qb and Qg. PMID- 16363315 TI - Sedative clearance during extracorporeal membrane oxygenation. AB - The effects of polyvinyl chloride (PVC) tubing and membrane oxygenator (MO) on the concentrations of lorazepam and morphine in the neonatal extracorporeal membrane oxygenation (ECMO) circuit were evaluated using an in vitro model that included a closed ECMO circuit with a MO, heat exchanger, bladder and PVC tubing. The circuit was primed with blood, electrolytes, albumin and heparin and maintained at physiologic pH and temperature throughout by frequent measurement of blood gas pH and a temperature probe. Lorazepam and morphine were each studied separately in three separate, but identical circuits for 6 h on the day of circuit prime (new circuit) and then again for 6 h at 24 h (old circuit). Each circuit (new and old) was spiked once with lorazepam to a final concentration of 250 ng/mL or with morphine to a final concentration of 70 ng/mL in the circuit. Serial samples were drawn at baseline and every 30-60 min for 6 h at the site of injection and pre- and post-MO for each circuit. Lorazepam and morphine concentrations were analysed using gas chromatography with electron capture and gas chromatography with mass spectrometry, respectively. The concentrations of morphine and lorazepam at various sample sites and time points were expressed as a percentage of the original concentration. This single-dose study shows that up to 50% of a dose of lorazepam and 40% of a dose of morphine may be extracted by PVC and MO during bypass, depending on the age of the circuit. As the circuits become older, this amount could increase. These data may explain, in part, the higher doses of lorazepam and morphine required to sedate patients during ECMO. PMID- 16363316 TI - Acute renal failure following open heart surgery: risk factors and prognosis. AB - BACKGROUND: Acute renal failure (ARF) development after cardiac surgery carries high mortality and morbidity. METHODS: Out of 14437 consecutive patients undergoing open-heart surgery between January 1991 and May 2001, 168 (1.16%) developed postoperative ARF mandating hemodialysis. Possible perioperative risk factors, and the prognosis of this dreadful, often fatal complication were investigated. RESULTS: The mortality rate in this group was 79.7% (134 patients). The risk factors associated with postoperative ARF were advanced age (p=0.000), diabetes mellitus (p=0.000), hypertension (p=0.000), high preoperative serum creatinine levels (p=0.004), impaired left ventricular function (p= 0.002), urgent operation (p=0.000) or reoperation (p=0.007), prolonged cardiopulmonary bypass (CPB) (p =0.000) and aortic cross-clamp (ACC) (p =0.000) periods, level of hypothermia (p =0.000), concomitant procedures (p =0.000), low cardiac output state (p =0.000), re-exploration for bleeding or pericardial tamponade (p =0.000), and deep sternal or systemic infection (p = 0.000). Of those who could be discharged from hospital, renal functions were restored in 21 patients (12.5%); however, eight patients (4.7%) became hemodialysis dependent. The mean follow-up period was 5.7+/-3.2 years (range: 4 months to 13 years; a total of 195 patient-years), and 10-year survival was 58.6+/-10.2% in the discharged patients. CONCLUSIONS: ARF development after cardiac surgery often results in high morbidity and mortality. Recognizing risk factors permits the timely institution of proper treatment, which is the key to reducing untoward outcomes. PMID- 16363317 TI - A novel method of measuring cardiac output in infants following extracorporeal procedures: preliminary validation in a swine model. AB - In infants, technologies for obtaining rapid, quantified measurements of cardiac output (CO) following weaning from cardiopulmonary bypass (CPB) or extracorporeal membrane oxygenation are not readily available. A new technique to measure CO based on ultrasound velocity dilution is described. It utilizes reusable probes placed on the extracorporeal circuit that permits convenient measurement of CO prior to decannulation. This report provides preliminary validation data in an animal model. Three Yorkshire pigs (11-14 kg) were fully heparinized and cannulated via the right common carotid artery (cannula advanced to the aortic arch) and right atrium. Both the venous and arterial lines were instrumented with ultrasonic probes connected to a computer-monitoring system. A 'stopcock bridge' between the arterial and venous cannulas provided the access for saline injection and a controlled AV-shunt. For comparison, a vascular flow probe was fitted directly to the pulmonary artery (PA) in both animals and, for the larger animal, a PA catheter was inserted to obtain standard thermodilution measurements. Linear regression analysis revealed a correlation between the CO by ultrasound dilution (CO UD) technique and the vascular probe and PA thermodilution techniques to be R2 =0.94 and 0.81. This pilot study demonstrated that the CO UD technique correlates to other benchmarks of CO measurements. This novel technology has specific application in the field of pediatric open heart surgery in that it would allow the surgeon to accurately and inexpensively measure the CO of neonatal and pediatric patients before and after surgical manipulation of the heart without the need for placement of additional catheters or probes. PMID- 16363318 TI - Comparison of conventional extracorporeal circulation and minimal extracorporeal circulation with respect to microbubbles and microembolic signals. AB - The intention of minimal extracorporeal circulation (MECC) is to reduce priming volume and minimize contact of blood with polymers and air in a closed system. In contrast to conventional extracorporeal circulation (ECC), a venous reservoir is missing. Thus, air trapping is limited and avoidance of bubble embolism is a major concern. This study investigates microbubbles (MBB) number and size in the venous and arterial lines of ECC and MECC compared to the number of microembolic signals (MES) in the right and left middle cerebral artery (MCA). Twenty patients undergoing coronary surgery were operated either with conventional ECC (cardiotomy reservoir, Rotaflow pump, Quadrox oxygenator, Quart filter) or MECC (Quart filter, Rotaflow pump, Quadrox oxygenator). Number and size of MBB were monitored in the venous and arterial lines with an ultrasound Doppler system. MES in right and left MCAs were measured by transcranial Doppler (TCD) monitoring. Patients undergoing MECC had additional sealing of the venous cannula by a ligature at the site of its insertion into the right atrium. There were no significant differences between groups with respect to age, X-clamping, bypass time and number of distal anastomoses. The number of MES and MBB in the arterial line was comparable between the groups. On the venous side, MECC-perfusion shows a significantly lower number of MBB. This could be explained with the additional sealing of the venous cannula. Furthermore, our data indicate that the MBB-volume reaching the pump will also appear in the arterial outflow and into the patient's MCA. For this reason, the avoidance of air contamination is a major concern for surgeons, anaesthesiologists and perfusionists. PMID- 16363319 TI - Leukocyte removal efficiency of cell-washed and unwashed whole blood: an in vitro study. AB - Leukocyte filtration of the cardiopulmonary bypass (CPB) perfusate after cardiac surgery has evolved as an important technique to prevent effector functions mediated by activated leukocytes. However, little is known about the filtration efficiency. Therefore, an in vitro study was conducted to define the leukocyte removal rate of a transfusion leukocyte-depletion filter, using cell-washed and unwashed whole porcine blood. In addition, the influence of different cell washing protocols on the elimination rate of blood cells (leukocytes and platelets) was investigated. Fresh, diluted, pooled, heparinized, porcine blood was processed using either a high-flow (HF, n = 5) or quality-wash (QW, n = 5) protocol on a continuous auto-transfusion system, or was left unprocessed (control n=5). Thereafter, all samples were filtered using a transfusion leukocyte-depletion filter. Blood samples for measurement of hematocrit, white blood cell count, including leukocyte differentiation and platelet count, were taken before and after filtration. To compare the experimental groups, the removal rate was presented as the fraction of leukocytes or platelets removed per plasma volume. Cell washing significantly altered the fraction of leukocytes removed per plasma volume when compared to unprocessed blood (2.07 and 2.36 in the HF and QW groups, respectively, versus 1.34 in the control group, p = 0.008 for both). No statistically significant difference in leukocyte removal rate was observed between the different cell-washing protocols. The leukocyte differential count showed that, during all experiments, the neutrophils were removed most efficiently (99.7%). Overall, significantly more platelets were depleted after cell washing compared to the control group (1.47 and 1.60 in the HF and QW groups, respectively, versus 1.12 in the control group, p =0.008 and 0.032, respectively). Furthermore, the amount of blood that could be filtered using a single pass technique did not significantly differ between the experimental groups. However, a larger variation in the total amount of filtered blood was observed in the unprocessed group (570+/-398 mL) compared to the cell-washed groups (360+/-42 and 430+/-97 mL in the HF and QW groups, respectively). In conclusion, blood processing with an auto-transfusion system significantly enhances the leukocyte and platelet removal efficiency of the transfusion leukocyte-depletion filter that was studied. In particular, neutrophils were efficiently removed. PMID- 16363320 TI - A new practical technique to reduce allogeneic blood exposure and hospital costs while preserving clotting factors after cardiopulmonary bypass: the Hemobag. AB - Recent data independently linking allogeneic blood use to increased morbidity and mortality after cardiopulmonary bypass (CPB) warrants the study of new methods to employ unique and familiar technology to reduce allogeneic blood exposure. The Hemobag allows the open-heart team to concentrate residual CPB circuit contents and return a high volume of autologous clotting factors and blood cells to the patient. Fifty patients from all candidates were arbitrarily selected to receive the Hemobag (HB) therapy. A retrospective control group of 50 non-Hemobag (NHB) patients were matched to the HB group patient-by-patient for comparison according to surgeon, type of procedure, age, body surface area (BSA), body weight and CPB time. Many efforts to conserve blood (Cell Saver and ANH) were employed in both groups. Post-CPB cell washing of circuit contents was additionally employed in the control group. There were no significant differences between the HB and NHB groups in regard to patient morphology, pre-op cell concentrations, distribution of surgeon or procedures (41% valve, 16% valve/coronary artery bypass graft (CABG), balance CABG), pump and ischemic times and Bayes National Risk scores. The average volume returned to the patient from the HB was 817+/-198 mL (1 SD). Average processing time was 11 min. The Hemobag contained an average platelet count of 230+/-80 K/mm3, fibrinogen concentration of 413 +/- 171 mg/dl, total protein of 8.0+/-2.8 gm/dl, albumin of 4.4+/-1.2 gm/dl and hematocrit of 43+/-7%. Factor VII, IX and X levels in three HB contents averaged 259% greater than baseline. Substantial reductions were achieved in both allogeneic blood product avoidance and cost to the hospital with use of the HB. Infusion of the Hemobag concentrate appears to recover safely substantial proteins, clotting factor and cell concentration for all types of cardiac procedures, maintaining the security of a primed circuit. PMID- 16363321 TI - PVC-plasticizer DEHP in medical products: do thin coatings really reduce DEHP leaching into blood? AB - The hemocompatibility of artificial surfaces in extracorporeal blood circulation systems can be improved by coatings. According to the literature, heparin coatings should avoid the leaching of the plasticizer di(2-ethylhexyl) phthalate (DEHP) into the blood from components made from plasticized polyvinyl chloride (PVC). DEHP and its metabolites are known to impair the fertility of male rodents; effects on human fertility are assumed. Three different surface coatings with and without heparin were examined in a Chandler Loop model at 37 degrees C using fresh human blood to evaluate their hemocompatibility and barrier property to plasticizer. The levels of toxic oxidation products of DEHP generated in the blood, particularly, were found as high as in the uncoated tubing. The coatings improved the hemocompatibility, but are not safe protection against the hazardous metabolites of DEHP. For pregnant women, neonates and children, we would recommend using the available surface-coated plasticized PVC tubing sets, but free of DEHP. PMID- 16363322 TI - Cardiopulmonary bypass and mitral valve replacement during pregnancy. AB - Gravid patient cardiopulmonary bypass remains a high-risk procedure with regard to fetal preservation. Maternal mortality is similar to that of the nonpregnant female at 1.5-5%. However, fetal mortality remains high at 16-33%, with an average of 19% over the past 25 years, with no correlation to gestational age. Teratogenesis is a major consideration in the first trimester. Variations in the timing of surgical intervention, gestational age, maternal health status, type of procedure, pre- or postorganogenesis, perfusion protocol, and pharmaceutical therapy are all factors that can influence fetomaternal outcome. In this report, we present a literature review along with our experience of a 26-year-old female who developed complications with her pregnancy at approximately 17 weeks gestation, with adverse neurological sequelae. The patient was 152 cm in height and weighed 48 kg, with a calculated body surface area of 1.40 M2. She had no prior history of cardiac disease and, upon admission to our institution, presented with a declining health status in pulmonary edema and was treated medically, with an ultimate requirement for mitral valve replacement. The total cardiopulmonary bypass time was 99 min with an aortic crossclamp time of 83 min. The literature, as expected, is limited to case reports and reviews since a controlled clinical trial during pregnancy is nonexistent, using extracorporeal circulation. This greatly challenges the medical staff in managing such difficult cases, with an incidence of heart disease during pregnancy of 1.2-3.7%. PMID- 16363323 TI - Just how pathogenic to pets and humans are the bacteria in the concentrations found in raw pet foods. PMID- 16363324 TI - How should the CVMA help developing countries? PMID- 16363325 TI - An ethicist's commentary on the case of the veterinarian reprimanded for poor records in a cruelty case. PMID- 16363326 TI - Effects of propofol and fentanyl on duodenal motility activity in pigs. AB - To investigate the effects of propofol and fentanyl on the postprandial duodenal motility the intraluminal impedance technique was used. Six pigs were instrumented with a central venous catheter, a percutaneous enterogastrostomy (PEG), and an impedance catheter, which was introduced via the PEG into the duodenum through endoscopy. Over the following 3 d, duodenal motility was measured for 8-hour periods. Measurements were taken on each subject under 3 different sets of conditions: in the conscious unrestrained pig, during propofol sedation, and during sedation with propofol-fentanyl. Both, after morning feeding and during gastric nutrition via the PEG, duodenal feeding patterns and duodenal phase II of the migrating motor cycle were shortened during propofol and propofol fentanyl sedation. In contrast, the duration of phase I was prolonged by propofol and propofol-fentanyl. In conclusion, either propofol or propofolfentanyl sedation shortens duodenal feeding patterns, as well as phase II of the migrating motor cycle. PMID- 16363327 TI - Efficacy of vaccination against Fusobacterium necrophorum infection for control of liver abscesses and footrot in feedlot cattle in western Canada. AB - A randomized and blinded field trial was carried out to evaluate the efficacy of a Fusobacterium necrophorum bacterin for control of liver abscesses and footrot under commercial feedlot conditions in western Canada. Half of the vaccinated and half of the unvaccinated control animals had ad libitum access to a forage-based (ALF) growing diet. The other half of each group was limit-fed a grain-based (LFG) growing diet. The overall prevalence of A and A+ liver abscesses in this trial was 16.7%. A strong association was found between diet group and presence of A or A+ liver abscessation at slaughter. Diet group modified the effect of vaccination on the prevalence of liver abscesses at slaughter, and on the incidence of footrot during the feeding period. The odds that a vaccinated animal in the ALF group would have an A or A+ liver abscess at slaughter were less than 1/3 the odds that an unvaccinated animal in the same diet group would have an A or A+ liver abscess at slaughter (OR = 0.27, [95% CI: 0.07 to 1.02], P = 0.05). The overall incidence of footrot in this trial was 6.5%. The odds that a vaccinated animal in the ALF group would be treated for footrot were less than 1/5 the odds that an unvaccinated animal in the same group would be treated for foot-rot (OR = 0.18, [95% CI: 0.04 to 0.82], P = 0.03). Within the LFG group there were no differences between vaccinated and unvaccinated animals in the odds of an animal being treated for footrot, or in the odds of having an A or A+ liver abscess score at slaughter. This trial suggests that vaccination against F. necrophorum infection may have applications to decrease the prevalence of severe liver abscesses at slaughter and decrease footrot treatments in certain diet situations. PMID- 16363328 TI - The effect of gastrointestinal parasitism on blood copper and hemoglobin levels in sheep. AB - Endoparasitism is a problem in Trinidad, as it is in most tropical countries. Parasite infection has been suspected to contribute to the pathogenesis of swayback disease (which is also prevalent in Trinidad), but the mode of action has not been clearly defined, although it has been suggested that parasites interfere with the absorption of copper from the gastrointestinal tract. The objectives of the study were to assess the effect of endoparasitism on blood copper levels and hemoglobin (Hb) concentration in sheep in Trinidad. Copper was administered intramuscularly to parasite infected and noninfected animals. The results showed that parasitism has a depressing effect on blood copper and Hb levels, even when administered parenterally. It is concluded that parasitism can aggravate existing hypocupremia and possibly influence the expression of swayback disease. PMID- 16363329 TI - [A clinical case of hypospodias in a dog]. AB - A 1-year-old intact male West Highland White Terrier was presented for abnormal miction. A clinical diagnosis of perineal type of hypospodias was established. Establishment of the diagnosis and the surgical management are discussed. PMID- 16363330 TI - Ovarian vascular hamartoma in a cow. AB - A 5-year-old Holstein cow was examined for a unilateral enlarged ovary. A clinical diagnosis of ovarian tumor was established before a histological diagnosis of ovarian vascular hamartoma was made. Ovarian vascular hamartoma has not been reported previously in a live animal. PMID- 16363331 TI - Ovarian teratoma and endometritis in a mare. AB - An 8-year-old Arabian mare was admitted for a large ovarian anovulatory follicle. A clinical diagnosis of ovarian tumor and endometritis was established. Histological examinations revealed an ovarian teratoma and a grade II endometritis. Three months after unilateral ovariectomy, the mare was confirmed pregnant and eventually gave birth uneventfully. PMID- 16363332 TI - Eosinophilic keratitis and keratoconjunctivitis in a 7-year-old domestic shorthaired cat. AB - A 7-year-old, spayed female, domestic shorthair was presented with a several month history of an ulcerated right cornea with marked stromal edema and inflammation that was nonresponsive to therapy. A diagnosis of eosinophilic keratitis was made, based on cytologic examination of corneal scrapings. The lesion resolved after treatment with topical corticosteroids. PMID- 16363333 TI - Disaster recovery plan. PMID- 16363334 TI - Hot iron branding--not a reasonable requirement for international trade in live ruminants. AB - "Brand" means a permanent identification mark of which the letters, numbers and figures used are each three (3) inches or more in length or diameter and are humanely burned into the hide of a live animal with a hot iron or tattoo or caustic chemical substance and is to be considered in relation to its location on the animal and the term relates to both the mark and location. PMID- 16363335 TI - Soft tissue sarcomas in dogs. PMID- 16363336 TI - Diagnostic ophthalmology. Right corneal foreign body, secondary ulcerative keratitis, and anterior uveitis. PMID- 16363337 TI - Respiratory syncytial virus epidemiology in Turkey. AB - Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infections in infants and young children worldwide. This study was conducted to determine the prevalence of RSV among high-risk children admitted with respiratory symptoms in a developing country. This is a multicenter study conducted among children less than 24 months of age and admitted to the hospital with respiratory symptoms. The inclusion criteria included: lower respiratory tract symptoms on admission, gestational age less than 35 weeks, and admission age less than six months, or children less than 24 months of age with a diagnosis of bronchopulmonary dysplasia requiring medical treatment or intervention during the last six months or with an uncorrected congenital heart disease (other than patent ductus arteriosus). Nasopharyngeal samples were obtained with one of the three standard methods: nasopharyngeal aspirate, nasopharyngeal wash or nasopharyngeal swab. RSV antigen was determined by enzyme immunoassay using Abbott TESTPACK RSV (No. 8100/2027-16). Statistical analysis was performed using Student's t-test and chi-square test. In this study, 332 children (135 females, 40.7%; 197 males, 59.3%) were included, and the nasopharyngeal specimens of 98 (29.5%) children were determined to be RSV-positive. There were no differences in sex, age of gestation, age of admission, family education, number of siblings and smoking at home for RSV-positive and -negative cases. Furthermore, underlying disease and duration of hospital and intensive care unit stay were similar among groups. Only otitis media was more common among RSV-positive cases. No fatality at hospital was recorded. Frozen samples revealed more negative results. Most cases presented during winter and the number of RSV-positive cases was higher in cold and economically poor areas. Premature children and children with underlying medical condition acquire RSV irrespective of other sociodemographic risk factors, and most of them are hospitalized. Thus, an RSV vaccine seems the most effective mode of protection to decrease morbidity and mortality. PMID- 16363338 TI - An epidemic caused by measles virus type D6 in Turkey. AB - In this study, the extent of measles outbreak was investigated in the Idil and Cizre counties of Sirnak Province. New cases determined in patients who applied to primary care clinics and those detected during home visits were evaluated. In 2001, a total of 2,143 cases reported in Sirnak Province were signified as a probable outbreak. Three hundred and thirty-three patients in Cizre and 219 patients in Idil applied to the primary care clinics. Of the cases, in Cizre 8.4% (n=28) and in Idil 6.4% (n=14) were infants aged nine months and younger who had not yet been vaccinated. Totally, 17 new cases (8 in Cizre and 9 in Idil) in the exanthema phase were determined during home visits and these were considered as outbreak cases. Virus isolation was achieved in 12 cases. All isolates were sent to the Centers for Disease Control (CDC) for genotyping and classified as D6 group. In conclusion, measles epidemics are still seen in our country. Therefore, measles outbreaks necessitate intensive intervention by physicians who are employed in primary health care services. PMID- 16363339 TI - Influenza and respiratory syncytial virus morbidity among 0-19 aged group in Yunus Emre Health Center. AB - The objective of the study was to determine the morbidity of influenza and respiratory syncytial virus (RSV) infection in the 0-19 years of age group with influenza-like illness among the outpatient cases. From 20 January to 31 March 2003 a total of 123 subjects with upper respiratory tract infection attended Yunus Emre Health Center. Ninety-one subjects fit the case definition of influenza-like illness, which consisted of acute fever of more than 38 degrees C, cough, and sore throat. After obtaining their consent, nasal swabs were taken for isolation of influenza and RSV. Of these, 10 were influenza A virus, 6 were influenza B virus and 20 were RSV. All of influenza virus A was typed as subtype H3N2. The rates of influenza virus among 5-9 and 1-4 years of age groups and of RSV among 1-4 years of age group were high. The average number of absentee days of schoolchildren with influenza was 3.33 days and of those with RSV infection was 1.43 days; this rate was calculated as 2.25 days for the influenza-like illness. Continuous surveillance and influenza vaccination for target groups are recommended for beneficial effects of reducing influenza morbidity and mortality in the community. PMID- 16363340 TI - Cardiac dysrhythmias after transcatheter closure of ASD with Amplatzer device. AB - Transcatheter closure of atrial septal defect (ASD) has been used as an alternative to open heart surgery. Although transcatheter closure of ASD with the Amplatzer septal occluder is a safe and feasible method in pediatric patients, there is little published data on arrhythmia analysis following transcatheter device closure of secundum ASD. We evaluated cardiac dysrhythmias with 24-hour ambulatory electrocardiographic (ECG) monitoring after transcatheter closure of ASD with Amplatzer device. A total of 85 consecutive patients with ASD underwent transcatheter closure of secundum ASD with Amplatzer device between October 1998 and December 2003. The study involved 65 of these patients assessed by 24-hour ambulatory ECG monitoring. Seven patients were evaluated a second time by 24-hour monitorization. During the procedure, transient complete atrioventricular (AV) block was seen in two patients. One of them returned to normal sinus rhythm in catheterization lab and the other returned to normal sinus rhythm in two hours. Transient junctional rhythm was observed in another patient during the device placement. Twenty-four hour ambulatory ECG monitoring was performed on all patients after a mean four-month period (1-12 months). Holter recordings demonstrated rare supraventricular extrasystole in two patients, rare ventricular premature beats in two patients, and intermittent sinus arrest with sinus pause lasting <1.5 seconds in one patient, for a total of five patients (7.6%). In conclusion, dysrhythmias after transcatheter device closure of secundum ASD with Amplatzer device are rare and benign. We need further long-term follow-up to evaluate late dysrhythmias after the transcatheter device closure of secundum ASD. PMID- 16363341 TI - Analysis of MTHFR 1298A>C in addition to MTHFR 677C>T polymorphism as a risk factor for neural tube defects in the Turkish population. AB - Maternal folic acid intake in the periconceptional period is strongly related to reduction in recurrence and occurrence of birth defects involving the neural tube. Among the single nucleotide polymorphisms (SNPs) influencing the folate metabolism, the methylenetetrahydrofolate reductase (MTHFR) gene has been the one most exclusively studied. Many studies have reported significant association between MTHFR 677C>T and increased risk of neural tube defects (NTDs). Our previous study did not support this observation. The present study aimed to determine the prevalence of 1298A>C polymorphism in addition to 677C>T in the same Turkish population as a risk factor for NTDs. We genotyped case (95 offspring with NTDs, 80 mothers, 72 fathers) and control (93 healthy children) populations for MTHFR 677C>T and MTHFR 1298 A>C polymorphisms. The comparison demonstrated a significant increase in the 1298AA/677TT genotype frequency among mothers of offspring with NTDs (OR 5.23 [1.06-25.9]; p=0.067). The 677CT genotype was only 1.35 times higher than controls among mothers when 677C>T polymorphism was evaluated alone, while 677CT/1298AC in the current study demonstrated a 3.8 times increase in this risk. These observations led us to conclude that although not statistically significant, MTHFR 1298AC polymorphism might be a risk factor for the occurrence of NTDs in the Turkish population. PMID- 16363342 TI - Ten-year experience in management of diabetic ketoacidosis and ketosis: 140 episodes at pediatric age. AB - One hundred and forty episodes in 112 patients (58 boys) with diabetic ketoacidosis (96 episodes) and diabetic ketosis (44 episodes) were studied to elucidate the clinical and laboratory risk factors for altered level of consciousness at presentation and to analyze the outcome of a distinct protocol in the treatment of diabetic ketoacidosis. The patients were analyzed according to demographic data and clinical and laboratory findings at admission. The treatment protocol involved use of 0.45% sodium chloride (NaCl) in 2.5% dextrose as the initial fluid therapy following volume expansion. Dextrose content of the fluid was doubled once the serum glucose level fell below 250 mg/dl. The mean ages at presentation with diabetic ketoacidosis and ketosis were 10.3 +/- 4.4 and 10.2 +/- 4.0 years, respectively. Thirty-one percent of patients had altered consciousness at presentation. The level of consciousness correlated negatively with serum bicarbonate level (r=-0.485; p<0.001). A serum bicarbonate level below 15 mmol/L was a risk factor for altered consciousness. There was no correlation between effective osmolality and the level of consciousness. Serum effective osmolality above 320 mOsm/kg H2O did not appear to be a risk factor for altered consciousness. No mortality or any signs of clinical brain edema were observed in patients treated with the distinct treatment protocol. In conclusion, acidosis appears to be the major factor in the pathogenesis of altered consciousness at presentation. Serum effective osmolality does not seem to be a risk factor as suggested previously. Dextrose added to the infusion fluid early in treatment seems to prevent the development of brain edema, and this may be due to a protective effect of higher osmolality in the resultant solution. PMID- 16363343 TI - Risk factors for hospitalization in children with acute diarrhea: a case control study. AB - To determine the risk factors for hospitalization in 0 to 24-month-old children with acute diarrhea, a retrospective case-control study was done at the Diarrheal Diseases Training and Treatment Center, where all the cases are managed according to World Health Organization (WHO) criteria. For each hospitalized patient, the next two consecutive admissions were enrolled as the control group. Clinical history and detailed physical examination results on admission were obtained from the standard file for all patients. The two groups were found to be similar for age, sex, height, blood in stool, and duration of diarrhea on admission. However, more children in the hospitalized group had high purging rate, frequent vomiting, malnutrition, dehydration and fever, and were not breast-fed on admission compared to the control group in univariate analysis. In multiple logistic regression analysis, only high purging rate, presence of dehydration and absence of breast-feeding on admission were the statistically significant factors. Mothers should be advised to seek early medical attention if the child has high purging rate (more than 8/day) and to continue breast-feeding up to 24 months of age. Multi-center studies with a large number of cases should be done to clarify the possible risk factors for hospitalization in cases with acute diarrhea. PMID- 16363344 TI - Superoxide dismutase activity in colostrum, transitional and mature human milk. AB - Colostrum and mature human milk are rich sources of nutrients and contain biologically active molecules that are essential for specific antioxidant functions. The aim of the present study was to determine the activity of copper, zinc superoxide dismutase (CuZnSOD) and manganese superoxide dismutase (MnSOD) activity in different phases of lactation. Specific enzyme activity was determined in colostral milk (3rd-5th days after delivery), and in mature milk in the third week (15-20 days), and the fourth and seventh months of lactation. In the third week of lactation, the activity of CuZnSOD and MnSOD was significantly higher in comparison to the colostral phase. In the fourth month of lactation, the activity of both enzymes was suppressed, while in the seventh month of lactation the MnSOD activity was increased, and the CuZnSOD activity was not significantly changed. These findings show that the activities of superoxide dismutases significantly change during different phases of lactation. PMID- 16363345 TI - Pediatric obstructive sleep apnea syndrome and anesthetic management. AB - Sleep-related breathing disorders require special attention in children who spend a considerable time sleeping. Obstructive sleep apnea syndrome is characterized by episodes of upper airway obstruction during sleep. Symptoms include hyperactivity, enuresis, headache, failure to thrive, and increased respiratory effort and total sleep time. The most common cause is adenotonsillar hypertrophy. Coexisting diseases are obesity, neuromuscular and craniofacial anomalies, and Down's syndrome. Early diagnosis is important to minimize neurocognitive, cardiac and developmental complications. Polysomnography is the gold standard for diagnosis. Although the features of pediatric obstructive sleep apnea syndrome are distinctly different from that in adults, it may predispose to the adult type of the syndrome. As therapy concerns several surgical approaches as well as conservative techniques, anesthetic management calls for particular attention. Pre- and postoperative sedation must be performed cautiously and patients must be watched closely with respect to airway obstruction and hypoventilation. Difficult intubation must always be considered. PMID- 16363346 TI - Symptomatic and asymptomatic hypohidrosis in children under topiramate treatment. AB - Topiramate (TPM) has peculiar side effects such as speech difficulties, weight loss, oligohidrosis and hyperthermia. We present the frequency and severity of hypohidrosis in our patients under TPM treatment. One hundred and two patients treated with TPM were evaluated retrospectively. Five (8 months-15 years of age) of them experienced symptomatic hypohidrosis manifested with prolonged or intermittent fever. Pilocarpine iontophoresis sweat test had been performed on the five patients before they were managed, and no sweat had been collected in 4/5 cases. Of the 102 patients, 42 who started TPM treatment when the study was established were evaluated prospectively regarding oligohidrosis. First, they were questioned about whether hypohidrosis occurred after TPM. Of 42, 11 patients complained of hypohidrosis. A sweat test was then performed on these 11 patients. Sweat could not be obtained in 5/11, and increased chloride concentration was found in 4/11. However, sweat could be obtained in the patients who had no complaint of hypohidrosis. Pediatricians should be aware of this side effect of TPM to prevent nonrelevant and cumbersome investigations in patients with prolonged or intermittent fever onset during TPM treatment. Our findings suggest that 5% of patients would experience hyperthermia during TPM treatment. Hypohidrosis without hyperthermia would be more frequent. If it is possible to collect sweat in patients who get fever during TPM treatment, it would be highly probable that the fever is not due to hypohidrosis. Increased chloride concentration alone does not seem to be significant for the hyperthermia risk. PMID- 16363347 TI - Role of scintigraphy in congenital thyroid anomalies. AB - Thyroid scintigraphy using Tc-99m pertechnetate is a frequently performed procedure in routine nuclear medicine practice. The indications for thyroid scintigraphy are investigation of hyperthyroidism, nodularity of the gland, cause of thyroid stimulating hormone elevation and localization of an ectopic thyroid gland. In the pediatric population, the most common request is for the evaluation of neonatal hypothyroidism. This imaging procedure is helpful in the identification of the underlying cause as well as in making a differential diagnosis. Early diagnosis is essential for appropriate therapy planning in this age group, and thyroid scintigraphy provides important diagnostic data. This article is written to review the scintigraphic findings in various congenital thyroid anomalies and to underline its use in the differential diagnosis. PMID- 16363348 TI - Atypical presentation of spondylitis in a case with sickle cell disease. AB - Salmonella osteomyelitis is rarely seen and vertebral involvement by this organism is very rare, especially in childhood. Salmonella should be considered in the list of etiologic agents of osteomyelitis in sickle cell disease. Herein we report a five-year-old boy with sickle cell disease and Salmonella paratyphi B spondylitis who was diagnosed with atypical clinical findings of infection. We would like to emphasize the importance of differentiation between osteomyelitis and infarction even if there is no obvious sign of infection in the sickle cell patient even at atypical localization. PMID- 16363349 TI - Use of alendronate in the treatment of vitamin D intoxication in infants. AB - Vitamin D intoxication remains a challenging problem due to lack of an efficient treatment. Bisphosphonates, inhibitors of osteoclast-mediated bone resorption, however, have been proposed as a safe and effective alternative in the treatment of vitamin D intoxication. We present here two infants with vitamin D intoxication who were successfully treated with alendronate. We propose that oral alendronate is an alternative to intravenous bisphosphonates in the treatment of vitamin D intoxication in infancy. PMID- 16363350 TI - Rickets-like radiological and biochemical features of neonatal mucolipidosis II (I-cell disease): report of two cases. AB - In this paper, two cases with mucolipidosis type II (I-cell disease) (proven in one presenting newborn and presumed in an elder deceased brother) are presented. These infants showed severe skeletal changes with diffuse periosteal new bone formation in long bones and ribs, marked osteopenia, and resorption of scapula, clavicula, and mandible. There was also irregular demineralization of metaphyses of long tubular bones, as seen in rickets. The activities of serum alkaline phosphatase and parathyroid hormone were markedly elevated. Phosphorus was decreased. Serum 1,25-dihydroxyvitamin D was slightly elevated, but 25 hydroxyvitamin D and calcium were normal. Dysostosis multiplex resembling rickets and very high alkaline phosphatase activity were due to defective osteoblastic activity, but the mechanism of elevated parathyroid hormone was not clear. We conclude that early skeletal manifestation of mucolipidosis type II is not clearly identified and that differentiation from congenital rickets or congenital hyperparathyroidism could be difficult. It is speculated that hyperparathyroidism in these patients could be related to the calcium-sensing receptor malfunction in the parathyroid gland. PMID- 16363351 TI - Mammary ductal ectasia as cause of bloody nipple discharge in a 28-month-old boy. AB - Bloody nipple discharge occurs rarely in infancy and may be secondary to mammary ductal ectasia. Discharge commonly resolves spontaneously, and ultrasonography is a useful diagnostic technique to detect the cause of discharge. We report a 28 month-old boy who presented with unilateral bloody nipple discharge for one month without signs of infection or mass. Ultrasound examination showed a dilated duct in the retroareolar region. No atypical cells were present on cytologic evaluation of the discharge. Further diagnostic studies were avoided and the discharge ceased completely one month later. We conclude that bloody nipple discharge is usually a benign and self-limited process in infancy and that it is advisable to avoid unnecessary invasive investigations initially. Invasive diagnostic studies or surgery should be reserved for cases with a palpable mass, persistent discharge or equivocal ultrasonographic findings. PMID- 16363352 TI - A neonate with malignant ectomesenchymoma. AB - Malignant ectomesenchymoma is a rare tumor reported in head-neck, abdomen and perineal regions. It consists of mesenchymal and neuroectodermal elements. In this tumor group, neoplastic cells are differentiated into neuronal cells. It also has at least one malignant mesenchymal element, generally rhabdomyosarcoma. In this report we present a neonate with ectomesenchymoma. PMID- 16363353 TI - Progressive familial intrahepatic cholestasis with normal GGT level appearing with lichenification and enlargement of hands and feet. AB - Progressive familial intrahepatic cholestasis is a serious disease of the liver, known as Byler disease, characterized by hepatocellular cholestasis. Severe pruritus and high serum bile acid concentrations are the most important diagnostic criteria of this autosomal recessive inherited disease. Here, we present a five-year-old boy with lichenification and enlargement of hands and feet as a sign of progressive familial intrahepatic cholestasis due to severe pruritus. PMID- 16363354 TI - Variable expressivity of congenital longitudinal radial deficiency and spinal dysraphism in monozygotic twins. AB - We report monozygotic twins with longitudinal radial hypoplasia and low (L5) spina bifida occulta, but with differing severity. There is only one previous report of similar twins. We report these identical twins with the expression of longitudinal radial hypoplasia with mirror image pattern on contralateral sides, and the association of low spina bifida occulta. PMID- 16363355 TI - Prolonged cellulitis due to plant thorn. AB - We report the case of a five-year-old healthy boy who was admitted with abscess and soft tissue infection between the left wrist and the distal end of the forearm. Postero-anterior X-ray examination of the forearm showed a transverse line on the radius; however, further comparative radiographic examinations of the forearm were not compatible with fracture. Enterobacter cloacae was identified in the pus culture and initial intravenous empiric treatment with sulbactam ampicillin was continued. Although the microorganism was susceptible to the empiric antimicrobial agent, at the eighth day of the treatment, inflammatory drainage was still present so further evaluations were performed. Ultrasonography of the forearm and wrist revealed only collection; magnetic resonance imaging showed a foreign body on the anterior distal section of the left forearm. The patient underwent operation and a 22 mm wooden foreign body was removed. Detailed history after removing a tree branch particle revealed that the boy had recently fallen from a tree causing an abrasion over the left wrist. The postoperative course was uneventful and clinical response was excellent in two days. In view of this case report, we would like to emphasize the importance of medical history and imaging studies in cases of cellulitis that do not respond to appropriate antimicrobial therapy. Another point to be kept in mind is that Enterobacter cloacae, which is very rarely reported as a causative agent for cellulitis, could be the etiological agent in cases after plant thorn injuries. PMID- 16363356 TI - Mumps hydrocephalus ameliorated with external drainage. AB - Mumps is still one of the most common childhood diseases in areas where national immunization has not been implemented. Although central nervous system manifestation is not so uncommon, hydrocephalus secondary to mumps is very rare. In this report, we present a toddler who developed severe hydrocephalus during mumps infection, which resolved via timely cerebral spinal fluid (CSF) drainage. We would like to emphasize early intervention for CSF external drainage. PMID- 16363357 TI - Importance and functions of European grasslands. AB - The European agricultural policy is not simple and needs to accommodate also social and environmental requirements. Grassland will continue to be an important form of land use in Europe, but with increased diversity in management objectives and systems used. Besides its role as basic nutrient for herbivores and ruminants grasslands have opportunities for adding value by exploiting positive health characteristics in animal products from grassland and through the delivery of environmental benefits. In fact grasslands contribute to a high degree to the struggle against erosion and to the regularizing of water regimes, to the purification of fertilizers and pesticides and to biodiversity. Finally they have aesthetic role and recreational function as far as they provide public access that other agricultural uses do not allow. But even for grassland it is very difficult to create a good frame for its different tasks (1) the provision of forage for livestock, (2) protection and conservation of soil and water resources, (3) furnishing a habitat for wildlife, both flora and fauna and (4) contribution to the attractiveness of the landscape. Nevertheless it is the only crop, able to fulfil so many tasks and to fit so many requirements. PMID- 16363358 TI - Biodiversity and agro-ecology in field margins. AB - This multidisciplinary study investigates agro-ecological functions (nature conservation, agriculture, environment) and implications of newly created, mown sown and unsown field margin strips installed on ex-arable land to increase biodiversity. From conservational concern, the development of species rich field margin strips was not strongly affected by the installed type of margin strip since species diversity converged over time, whether strips were sown or not. Convergence between unsown and sown margin strips occurred also in terms of species composition: unsown and sown strips became similar over time. Mowing without removal of cuttings significantly reduced species richness, yielded more grassy margin strips and delayed similarity in species composition between sown and unsown margin strips. Species richness on the longer term was not significantly affected by light regime nor by disturbance despite significant temporary effects shortly after the disturbance event. On the contrary vegetation composition in terms of importance of functional groups changed after disturbance: the share of spontaneous species within functional groups increased resulting in higher similarity between the sown and unsown vegetation. Furthermore risk of invasion was highest in the disturbed unsown community on the unshaded side of a tree lane. A positive effect of botanical diversity on insect number and diversity was found. However the effects of botanical diversity on insect number was mediated by light regime. At high light availability differences between plant communities were more pronounced compared to low light availablilty. The abundance of some insect families was dependent on the vegetation composition. Furthermore light availability significantly influenced insect diversity as well as the spatial distribution of families. From agricultural concern, installing margin strips by sowing a species mixture and a mowing regime with removal of cuttings are good practices to diminish the risk of species ingrowth into adjacent crops by creeping roots and rhizomes. Seed dispersal was only problematic one year after the installation of the field margin strips particularly nearby the unsown margin strip and wind-borne seeds were dispersed over limited distances, mainly within 4 m of field margins. Annual herbage yield was not affected by mowing management. DM yield of sown/unsown communities converged over time. Compared to herbage from an intensively managed fertilized grassland, field margin herbage revealed a low feeding value, owing to a low crude protein content, a low digestibility and a high crude fibre content. The unsown community had a higher forage quality than communities sown to bred commercially available grass varieties. Both digestibility and crude protein content decreased over time irrespective of plant community or location. Mid June cuts were more productive than mid September cuts but digestibility and crude protein content was lower. The use of herbage from field margins as hay for horses or as a component in farmland compost are good alternatives. A strong relation was found between the distribution of pest insects and their antagonist families along field margin strips indicating a status of biological equilibrium. From environmental concern, field margin strips buffered boundary vegetation and watercourses against cropped areas loaded with high levels of mineral nitrogen. Margin strips reduced the mineral nitrogen content of the soil in the margin and mineral nitrogen loss during wintermonths. Mineral nitrogen loss was not affected by field margin type but by distance from the field crop. A minimal width of 5 m is necessary to reach an optimal reduction in mineral soil N and N losses. PMID- 16363359 TI - Production of auxin and related compounds by the plant parasitic nematodes Heterodera schachtii and Meloidogyne incognita. AB - Mass spectrometric analysis revealed the presence of auxin, mainly in conjugated form, in secretions of Heterodera schachtii and Meloidogyne incognita, with or without treatment with DMT or resorcinol. M. incognita showed the highest production rates, though treatment of M. incognita with resorcinol had a negative effect on auxin production. Analysis of auxin precursor molecules in lysates of H. schachtii, M. incognita and Caenorhabditis elegans suggested that auxin is most probably a degradation product of tryptophan and that auxin may be synthesized via several intermediates, including indole-3-acetamide which is an intermediate of a pathway so far only characterized in bacteria. Furthermore, high levels of anthranilate, a degradation product of tryptophan in animals, but possibly also a precursor for auxin were detected. PMID- 16363360 TI - Effect of plant density on competitiveness of Brassica napus, Sinapis alba and S. arvensis under water stress conditions. AB - Under Mediterranean climate, oilseed rape is subjected especially to the competition of weeds with respect to water. Herbicides registered for this crop do not effectively control species of the same family, in particular Sinapis alba and Sinapis arvensis. Moreover, there are no results of the effect of plant density on the competitiveness of these species. The purpose of this experiment was to determine if the competitiveness of the species varies according to the total density. The experiment was carried out in pots under greenhouse conditions, according to a replacement series method. Plant densities tested were 2, 4 and 8 plants per pot. The results of the replacement series diagram and those of relative crowding coefficients showed that Brassica napus was the most competitive, whatever the density is. This classification is explained primarily by leaf area. Indeed, the intraspecific competition due to B. napus has affected more its leaf area than the interspecific competition. Conversely, the intraspecific competition due to S. arvensis has less affected its leaf area than the interspecific competition. Regarding S. alba, the intraspecific competition effect was less severe than the interspecific competition effect due to B. napus and more severe than the interspecific competition effect due to S. arvensis on S. alba PMID- 16363361 TI - Competitiveness and dry matter allocation of oilseed rape (Brassica napus L.) and two mustards (Sinapis alba L. and S. arvensis L.) under water stress conditions. AB - In Morocco, oilseed rape is commonly exposed to mustards competition which are not totally controlled by herbicides. To understand the competitiveness of each species, growth parameters should be studied notably dry matter allocation. The objective of this study was to confirm the competitiveness of oilseed rape with regard to Sinapis alba and S. arvensis and to investigate how the dry matter is allocated. A pot experiment was undertaken with a quartz sand as substrate. Two plant densities were tested (one and two plants). The binary density was either a monoculture or a mixture. Half the pots were maintained at field moisture capacity and the other half was irrigated up to 70% of its water holding capacity. Dry matter allocation of each species at density two was compared to that of the same species at density one. Results of replacement series diagrams and those of the relative crowding coefficient (based on total dry matter) showed that Brassica napus was more competitive than S. alba. S. arvensis was the least competitive. Under competition, B. napus irrigated at water holding capacity allocated more dry matter to stem when compared to its dry matter at density one. Under the same condition, when reducing water supply, B. napus allocated more dry matter to leaves. In case of S. alba, dry matter percent in leaves and roots were respectively low and high in normally irrigated plant and inversely under water shortage. S. arvensis allocated high and low dry matter percent respectively to root than to leaves when sufficiently irrigated. But no clear tendency was noticed under water shortage, for this species. PMID- 16363362 TI - Unexpectedly high susceptibility of micropropagated rhubarb (Rheum rhaponticum L.) to spot disease caused by Ramularia rhei. AB - Rapid, large-scale micropropagation of a Rheum rhaponticum (rhubarb) breeding line, selected for the absence of autumn dormancy, produces changes typical of somaclonal variation. Among these is enhanced susceptibility to foliar infection by the spot disease pathogen Ramularia rhei, with consequential increased spore production on infected leaves. This provides an increased level of inoculum in the late season, leading to commercially unacceptable petiole spotting as spores are washed from the lamina on to the petiole surface. Infection of conventionally propagated plants does not exceed market acceptability. Consequently, to maintain the commercial value of this rhubarb selection micropropagation must be avoided, at least until an acceptably modified protocol is available. PMID- 16363363 TI - Impact of the nematophagous fungus Pochonia chlamydosporia on nematode and microbial populations. AB - The microbial and nematode populations associated with two plants (tomato and cabbage) inoculated with the nematophagous fungus, Pochonia chlamydosporia var. chlamydosporia or root knot nematode (Meloidogyne incognita), or both, were compared with those in unplanted controls. The dominant factor affecting culturable microbial populations was found to be the presence or absence of tomato plants. Generally microbial colony counts were lowest in unplanted soil, small increases were associated with cabbage and significantly greater numbers with tomato plants. Differences in microbial diversity (estimated from community profiles of carbon substrate utlisation, using Biolog) were observed between planted and unplanted soils, however, there were few differences between soils with either of the two plants. The presence of P. chlamydosporia was associated with a reduction in the numbers of plant parasitic nematodes (51%-78%) including the migratory ectoparasites, whereas free-living nematodes, culturable bacteria and bacterial populations assessed by Biolog were unaffected by the application of fungus. PMID- 16363364 TI - [Increased nuchal translucency with normal karyotype]. AB - Increased fetal NT thickness is a common phenotypic expression of trisomy 21 and other chromosomal abnormalities, but it is also associated with fetal death and a wide range of fetal malformations, deformations, dysgenesis, and genetic syndromes. This paper reviews the outcome of chromosomally normal fetuses with increased NT On the basis of these data it is possible to estimate, for each NT group, the chances of intrauterine survival and delivery of a healthy baby with no major abnormalities. These data are useful in counselling parents of pregnancies with increased fetal NT and in planning the appropriate follow-up investigations. In normal fetuses NT thickness increases with fetal crown-rump length (CRL). The median and 95th centile of NT at a CRL of 45 mm are 1.2, and 2.1mm and the respective values at CRL of 84mm are 1.9 and 2.7 mm. The 99th centile does not change significantly with CRL and it is about 3.5 mm. Increased NT, refers to a measurement above the 95th centile and the term is used irrespective of whether the collection of fluid is septated or not and whether it is confined to the neck or envelopes the whole fetus. After 14 weeks, increased NT usually resolves but in some cases it evolves into nuchal edema or cystic hygromas. PMID- 16363365 TI - [Conjoined twins--diagnostic problems, difficult decisions in choosing the most optimal management in one of the rarest pathologies of pregnancy]. AB - Conjoined twins are the rarest type of monozygotic, monochorionic, monoamniotic twins--incidence about 1:50000 - 1:100000. A conjoined twin pregnancy is a very special pathology in obstetrics and usually ends prematurely. 40% of twins are stillborn, while 35% die within 24 hours after delivery. The development of ultrasonography and in the last years also MRI, allows earlier detection and diagnosis of fetal malformation and enables precise evaluation of the anatomy, type of malformation and how the twins are conjoined. Chances of survival and way of delivery depends on the degree of fusion of the organs and possibilities of surgical separation. Obstetric care for a women with conjoined twins does not differ from that in a normal twin pregnancy, but the newborns require special medical care after delivery. We would like to stress the diagnostic problems and the difficult decisions that we encountered. The degree of conjoinment in this case excluded surgical separation of the twins. A multidisciplinary team of experts decided to end the pregnancy at 31 wks by cesarean section. Delivery at a later term could increase the risk of uterine rupture and surgical complications. PMID- 16363366 TI - [Genetic polymorphism of the calcitonin receptor gene and bone mineral density in Polish population of postmenopausal women]. AB - INTRODUCTION: In recent years the influence of genetic factors in the pathogenesis of osteopenia and osteoporosis was indicated. The investigations focused on the gene coding for calcitonin receptor. The goal of our analysis was to determine the genotype frequencies of AluI polymorphism of the calcitonin receptor gene (CTR) in the group of Polish postmenopausal women and its possible contribution to osteoporosis development. MATERIAL AND METHODS: 139 postmenopausal women with osteopenia (t-score value from -1.0 to -2.5) (mean age 58.5 +/- 5.9 years, mean age of menopause 49.8 +/- 3.9 years) have been investigated. AluI polymorphism of the CTR gene was determined using PCR/RFLP assay. We have analysed 3 subgroups: CC, CT, and TT. In each subgroup mean weight, height, body mass index (BMI), mean age of menopause and years since menopause (YSM) and parameters of bone turnover: bone mineral density (BMD), t score, index: young adults (YA) and--age matched (AM) have been analysed. Additionally the group of 138 selected women (mean age 26.5 +/- 4.3 years) as general population has been analysed. RESULTS: In investigated group the frequency of all 3 genotypes was determined as follows: CC: CT : TT = 8.6 : 45.3 : 46.1. Analysing BMD in particular subgroups the higher value for the CT genotype (0.967 +/- 0.161 g/cm2) was found. Similarly t-score (-1.94), YA (80.6%) and AM (90.8%) index were higher in CT genotype carriers. CONCLUSION: Our results suggest possible connection of the AluI polymorphism of the CTR gene with osteopenia and osteoporosis development. To confirm this tendency further investigations in the large number population are necessary. PMID- 16363367 TI - [Assessment of thyroid function in women treated for vomiting in early pregnancy]. AB - OBJECTIVES: Hyperthyroidism does not influence procreation ability significantly, but very often causes complications of the course of pregnancy and increases neonatal morbidity and mortality in newborns. Nausea and vomiting in the first trimester of pregnancy are very common complaints in these patients. DESIGN: The aim of the study was to prove, that assessment of the thyroid hormones in every vomiting woman in the first trimester allows to start the proper treatment of hyperthyreosis as early as possible and improves the prognosis. MATERIALS AND METHODS: Authors analyzed 22 pregnant women treated for vomiting during the first trimester in the Department of Obstetrics and 5 women treated in the Outpatient Clinic. RESULTS: In none of them hormonal dysfunction was diagnosed before pregnancy. In 21 patients free thyroxine (fT4) in serum was assessed. In 8 cases (38%) hyperthyreosis was diagnosed. Except the routine anti vomiting treatment, they received a beta-blocker 30 mg per day. It allowed to avoid hospitalization in three pregnant women treated in the outpatient clinic. Two patients, who had earlier two spontaneous abortions, delivered healthy babies. CONCLUSIONS: 1. The beta-blocker in treatment gives very good results and allows to avoid hospitalization in patients with hyperthyroidism and hyper emesis gravidarum. 2. In some patients vomiting may be the only symptom of sub clinical hyperthyroidism in early pregnancy. Assessment of the thyroid hormones levels and adequate treatment may give these women a chance for a healthy child. PMID- 16363368 TI - [Total and ionized magnesium concentration in the blood plasma and erythrocytic magnesium concentration of women in the third trimester of pregnancy with imminent preterm labor]. AB - OBJECTIVES: There are some data concerning magnesium concentration influence on the risk of preterm labor. The estimation of magnesium concentration changes may be useful in prevention of preterm labor. DESIGN: Therefore the aim of our study was to find out the correlation between magnesium concentration and the risk of preterm labor. MATERIALS AND METHODS: Total magnesium concentration and ionized magnesium concentration in blood plasma and erythrocytic magnesium concentration ware examined in the three groups of: 23 women in the third trimester of pregnancy with imminent preterm labor under tocolytic therapy; 20 women in the third trimester of physiologic pregnancy and 19 non-pregnant healthy women in the reproductive age. RESULTS: We discovered statistically confirmed differences (p < 0,05 ) in ionized magnesium concentration as well between the group of women in physiologic pregnancy and non-pregnant women and between the group of pregnant women with imminent preterm labor and non-pregnant women. CONCLUSIONS: Although there were no statistically confirmed differences in total magnesium concentration and erythrocytic magnesium concentration between the three groups of examined women, there were statistically confirmed differences in ionized magnesium concentration between the pregnant and non-pregnant women. Our results suggest that ionized magnesium concentration is better indicator of magnesium balance in human's body than total magnesium concentration. PMID- 16363369 TI - [Risk assessment for preterm delivery in pregnant women with active periodontitis]. AB - INTRODUCTION: Premature labor has been regarded as a highly problematic clinical event not only due to its relation with perinatal mortality but also in the light of the remote results of the prematurity. Therefore many ongoing studies concentrate on evaluation of potentially avoidable risk factors of premature labor. Relatively few studies have suggested chronic periodontitis as such a determinant. OBJECTIVES: Prospective evaluation of periodontium changes in the course of otherwise normal pregnancy. Assessment of potential influence of periodontium pathology, and its severity, on the obstetrical complications. The authors have made literature search on the relation of active periodontitis and premature labor. MATERIALS AND METHODS: Study involved 120 randomized gravidae attending electively dental clinic for a routine control. INCLUSION CRITERIA: single pregnancy, no preterm delivery in a history, mother's age 18 to 35 years, gestational age 10 to 20 weeks based on a last menstrual period date--confirmed on ultrasound. Every patient underwent dental examination twice--at the inclusion date and 48 hours after delivery. Patients were divided into three groups. Group I (n = 25)--healthy patients. Group II (n = 61)--patients with mild or moderate periodontitis. Group III (n = 39) patients with a severe periodontitis. Progression of lesions was recognized if the post-delivery dental assessment revealed increase of the pockets depth higher than 2mm and in at least 4 places compared with the initial findings. RESULT: In the course of otherwise normal pregnancy women with signs of inflammation within periodontium revealed progression of this pathology proportional to its severity in early pregnancy. This increase was associated with 4-fold higher rate of preterm deliveries in Group II and 12-fold in Group III compared with healthy patients. CONCLUSION: Active and severe periodontitis should be regarded as independent and highly influential risk factor of the preterm delivery. PMID- 16363370 TI - [Quantitative computerized tomographic densitometry for the evaluation of osteoporosis risk in menopausal women]. AB - OBJECTIVES: The aim of the study was retrospective analysis of osteoporosis risk in menopausal women. DESIGN: Retrospective analysis of bone mineral density of 2733 women was performed between 1997 - 1999 year. MATERIAL AND METHODS: Retrospective analysis of bone mineral density of 2733 women aged 16 to > 68 by quantitative computer tomography densitometry of L2-L4 vertebrae. RESULTS: The most rapid bone mineral density loss concerns postmenopausal women aged 59 to 63. By 18.7% comparing to the group 54-58 years and 59-63. CONCLUSIONS: 82% women who undergo densitometry is in the group over 44 years old. Bone mineral density loss is depending on age and concerns postmenopausal women, mainly. PMID- 16363371 TI - [Epidermal growth factor (EGF) in uterine leiomyoma during tumour growth]. AB - OBJECTIVES: Extracellular matrix is a place where various growth factors are bound and immobilised. It is expected that leiomyoma-associated remodelling of extracellular matrix in the uterus may be evoked by changes in the content of some growth factors. DESIGN: The amount and distribution of EGF in the normal myometrium and in the leiomyoma during various growth stages were investigated. MATERIAL AND METHODS: The assay of EGF was carried out with the use of ELISA commercial kit. SDS/polyacrylamide gel electrophoresis of tissue extracts followed by Western immunoblot was performed. RESULTS AND CONCLUSIONS: It was found that all investigated tissues contained endogenous EGE Extractability of EGF depended on type of extracting solvent. Only slight amount of EGF could be extracted by 1 M acetic acid. Much more EGF could be solubilized in 0.05M Tris/HCl, pH 7.6. Our results showed that EGF bound to the uterus (normal and leiomyoma) components of different molecular mass. It is of interest that some components of both, acidic and neutral extracts (myometrium and leiomyomas) were not able to bind exogenous 125I-labelled EGF. PMID- 16363372 TI - [Twin pregnancy with single fetal death due to triploidy--a case report]. AB - The incidence of multiple pregnancies is associated with the increased risk in maternal and fetal complications. Intrauterine death of one twin in the second trimester is a rare obstetric complication. Authors report a case of a twin pregnancy with triploidy of one fetus and no chromosomal anomaly of the other twin in a dichorionic diamniotic twin pregnancy. Amniocentesis at 16th weeks disclosed triploidy of this fetus who died afterwards at 20th week of gestation. The pregnancy was continued with special care of the mother and the alive fetus. The second twin was successfully delivered by cesarean section in the 41st week of pregnancy due to the intrauterine fetal distress. PMID- 16363373 TI - [Fetal survival after umbilical cord prolapse for more than three hours. A case report]. AB - We report a case of a 43-year-old woman, gravida 4, para 3, diagnosed with umbilical cord prolapse for more than three hours. Due to severe variable decelerations and fetal bradycardia, after counseling the patient decided to deliver by cesarean section. The infant, a 800-g male, spent 3 months in neonatal intensive care unit. At the ten-month follow-up examination he was neurologically intact, with no long-term complications secondary to the cord prolapse. PMID- 16363374 TI - [Ocular changes during pregnancy]. AB - Pregnancy is often associated with ocular changes. Most of them are specifically associated with the pregnancy itself, rare disturb the normal visual function and are transient in nature. The ocular effects of pregnancy may be divided into physiologic changes, pathologic conditions or modifications of pre-existing conditions. Pregnancy is associated with changes in corneal sensitivity and thickness, decreased tolerance to contact lenses, decreased intraocular pressure, can affect refractive error. Pathologic conditions reported to develop during pregnancy include hypertensive and vascular disorders, central serous chorioretinopathy, uveal melanoma and others. The most significant modified pre existing condition is diabetes mellitus. Pregnant women are at increased risk for the progression of preexisting proliferative diabetic retinopathy, and diabetic women should see an ophthalmologist before pregnancy or early in the first trimester. This review describes various ocular changes that occur during pregnancy and summarizes the ocular disturbances in these three categories. PMID- 16363375 TI - [Pulsed dose brachytherapy--a review of clinical applications in the treatment of gynecological malignancies]. AB - Pulsed Dose Rate (PDR) treatment is a new brachytherapy modality that combines physical advantages of High-Dose-Rate (HDR) technology (isodose optimization, radiation safety) with the radiobiological advantages of Low-Dose-Rate (LDR) brachytherapy. Pulsed brachytherapy consists of using a stronger radiation source than for LDR brachytherapy and is giving a series of short exposures of 10 to 30 minutes in every hour to approximately the same total dose in the same overall as with the LDR. Modern afterloading equipment offers some advantages over interstitial or intracavitary insertion of separate needles, tubes, seeds or wires. Isodose volumes in tissue can be created flexibly by a combination of careful placement of the catheter and adjustment of the dwell times of the computerized stepping source. Automatic removal of the radiation sources into a shielded safe eliminates radiation exposures to staff and visitors. Radiation exposure is also eliminated to the staff who formerly loaded and unloaded a multiplicity of radioactive sources into the catheters, ovoids, tubes etc. This retrospective study based on summarized clinical investigations analyses the feasibility, reasons for introducing PDR brachytherapy, preliminary results of PDR brachytherapy in treatment of gynecological malignancies and proposed treatment schema. PMID- 16363376 TI - [An open letter to all Polish Scientific Societies, government, and non government organizations about a nationwide program for cervical cancer prevention]. PMID- 16363377 TI - [Recommendations of the Polish Society of Obstetrics and Gynecology for prenatal care in normal pregnancy]. PMID- 16363378 TI - [Sonographic features of chromosomal defects in the second trimester of pregnancy]. AB - Each chromosomal defect has its own syndromal pattern of detectable abnormalities. The authors describe the sonographic features of trisomy 21 and other major chromosomal defects in the second trimesters of pregnancy. It is recommended that, when an abnormality/marker is detected at routine ultrasound examination, a thorough check is made for the other features of the chromosomal defect known to be associated with that marker; should additional abnormalities be identified, the risk is dramatically increased. In the case of apparently isolated abnormalities, the decision whether to carry out an invasive test depends on the type of abnormality. Minor fetal abnormalities or soft markers are common and they are not usually associated with any handicap, unless there is an underlying chromosomal defect. Routine karyotyping of all pregnancies with these markers would have major implications, both in terms of miscarriage and in economic costs. It is best to base counseling on an individual estimated risk for a chromosomal defect, rather than the arbitrary advice that invasive testing is recommended because the risk is 'high? The estimated risk can be derived by multiplying the a priori risk (based on maternal age, gestational age, history of previously affected pregnancies and, where appropriate, the results of previous screening by NT and/or biochemistry in the current pregnancy) by the likelihood ratio of the specific abnormality or marker. PMID- 16363379 TI - [Diagnostic difficulties of Toxoplasma gondii infection in pregnant women. Is it possible to explain doubts by polymerase chain reaction?]. AB - OBJECTIVES: Toxoplasma gondii infection during pregnancy is still a difficult problem in the contemporary perinatology. Difficulties met during interpretation of serological tests carried out in pregnant patients to detect Toxoplasmosis implies more and more frequent use of the Polymerase Chain Reaction (PCR). DESIGN: To evaluate the dependence between serological tests and quantity of the Toxoplasma gondii genomes in mothers' blood and amniotic fluid or neonatal blood, the quantitative PCR (q-PCR) method was applied. MATERIALS AND METHODS: The analysis was performed in 81 pregnant women. Maternal blood, amniotic fluid and newborns' umbilical blood samples were evaluated for the presence of Toxoplasma gondii DNA. IgG and IgM Toxoplasma gondii antibodies were evaluated by the ELISA method. RESULTS: High seroprevalence (51.9%) of the Toxoplasma gondii was confirmed. Toxoplasma gondii genetic material in blood and/or amniotic fluid was found in 33 patients. It was stated that quantity of the protozoa and anti-IgM presence in mothers' blood are the factors influencing significantly the Toxoplasma gondii manifestation in amniotic fluid. CONCLUSION: High suitability of PCR in diagnosis of Toxoplasmosis during pregnancy and vertical transmission was confirmed. PMID- 16363380 TI - [Transvaginal ultrasonography of the cervix length and the prediction of preterm delivery]. AB - OBJECTIVE: To determine if cervical shortening between 12 and 28 weeks gestation predicts risk of spontaneous preterm birth or cervical funneling requiring cerclage. STUDY DESIGN: We reviewed retrospectively all the patients who had transvaginal cervical ultrasound assessment during one year. 43 patients who had a second and early third trimester delivery or cerclage placement were chosen for the study. The control group consisted of 78 patients without signs of preterm labor and cervical shortening as assessed by transvaginal ultrasound. The results of the last ultrasound cervical length measurement were correlated with the likelihood of preterm delivery or cervical funneling requiring cerclage. RESULTS: 26 of 121 patients studied delivered preterm and 15 required cervical cerclage because of cervical shortening and funneling. The mean cervical length assessed by ultrasound was 23.22 +/- 3.07 mm in the group with preterm contractions and delivery and 21.99 +/- 7.05 mm in the group with cervical incompetence. In the normal pregnancy group mean cervical length was 35.59 +/- 3.07 mm. CONCLUSIONS: Cervical length of < 3.0 cm and further shortening of the cervix is predictive of preterm delivery or cervical incompetence requiring cerclage. PMID- 16363381 TI - [Diagnosis of papilloma viruses in cervical cancer in 414 women from Wielkopolska region by the immunohistochemical assessment]. AB - OBJECTIVES: Cervical cancer in Poland is second most common type of cancer, after breast cancer. There are known risk factors of cervical cancer development and the most serious one is human papilloma virus infection (HPV). DESIGN: The aim of our paper is present the result study 414 cervical cancer of women from Wielkopolska region treated at the Department of Gynecology and Obstetrics, Karol Marcinkowski University of Medical Sciences in Poznan. MATERIALS AND METHODS: In morphological study grading, staging and vascular invasion were estimated. In cervical cancer tissue papillomaviruses PCR method were used. In immunohistochemical study, expression of proteins checking the cell cycle, share in signal transduction to nucleus, cell receptors for steroid hormones and viruses oncogenic proteins were investigated. In the part of cancer gene mutation of p53 (60 cancers) i k-RAS (40 cancers) were searched. RESULTS: In cancers HPV 16/18 infected vascular invasion were more frequently (p < 0.013). No statistically significant difference in cellular proteins expression in the HPV16/18 positive cancers, HPV16/18 negative and cancers without HPV was observed. However significant difference were demonstrated in proteins expression depending from degree of cancer stage. CONCLUSIONS: The result of these studies suggest that super expression for EGFr is poor prognostic factor in the early stage of cancers (I-II0). PMID- 16363382 TI - [The correlation between prognostic factors and outcome of second look laparotomy in ovarian cancer]. AB - OBJECTIVES: The objective of this study was to verify the correlation between prognostic factors, positive or negative second look laparotomy (SLL) and 5-year survival in patients with ovarian cancer. MATERIAL: Between 1984 and 1993, 178 patients after primary surgery and first-line chemotherapy with complete clinical response underwent second look laparotomy. The correlation between clinical stage and grade of cancers, residual disease, age of patients in two group of patients: with positive and negative second look laparotomy were evaluated. RESULTS: 95 patients (53%) had positive SLL. The 5-years survival in this group was only 20%. Adverse prognostic factors were: advanced primary stage (IV-0% of 5-years survival), low grade of differentiation (9.1% of 5-years survival) and residual disease > 2cm (9.4% of 5-years survival). The 5-year survival in group with negative SLL was 78.3%. The analysed prognostic factors in this group were insignificant in predicting 5-year survival. CONCLUSIONS: This study confirms that the SLL can provide an important prognostic evaluation in patients without evidence of disease and allows surgical cytoreduction in group with positive SLL (more than 50% of patients). The analyzed prognostic factors in group with negative SLL were insignificant in predicting 5-year survival. The multicenter research for new prognostic factors in this group are required. PMID- 16363383 TI - [The internal jugular vein thrombosis during pregnancy with decreased level of protein S]. AB - The study presents a rare case of internal jugular vein thrombosis during pregnancy. The patient's only complaints were headaches which increased after conception. In the 11th week of pregnancy thrombosis of the internal left jugular vein was diagnosed. Since then, patient had been treated with low molecular weight heparin. Further course pregnancy, delivery and puerperium were not complicated. The decreased level of protein S was responsible for the internal jugular vein thrombosis in this case. PMID- 16363384 TI - [CCAM type II with no clinical symptoms before and after birth--diagnosed in the 21st week of pregnancy and child pediatric observation in the first 3 years of age--a case report]. AB - CCAM is rare congenital malformation of the respiratory tract. Presented case of CCAM type II was diagnosed based on the prenatal examination. In presented case without clinical signs after delivery diagnosis of CCAM type II was established based on prenatal diagnosis. This allowed after delivery to take up focused radiological diagnostics and further proper malformation monitoring. PMID- 16363385 TI - [Meconium as a new biological material for detecting intrauterine exposure to toxic substances]. AB - The use of licit and illicit drugs and exposure to other xenobiotic agents during pregnancy is common. These substances are known to have adverse effects on the pregnancy and fetus; however information on fetal exposure is sparse due to the lack of an appropriate measure of exposure. Meconium analysis is a new method for identifying in utero exposure of infants to a number of illicit and legal drugs, alcohol, nicotine, heavy metals, pesticides, congenital infections. It's testing is non-invasive, highly accurate and able to detect prior exposure in utero during 12-40 weeks of gestation. This has implications for toxicology to develop improved methods to identify exposed infants. PMID- 16363386 TI - [The role of insulin resistance and resistin in pathogenesis of diabetes mellitus with special attention to diabetes in pregnancy]. AB - During pregnancy, energy metabolism is altered to meet the increased energy demand by the rapidly growing fetus. Postprandial hyperglycemia is usually observed in gestation due to the decreased insulin sensitivity. The placenta derived hormones are partially responsible for the insulin resistance. The mechanism of this event in still unclear. The results of experimental studies suggest that resistin, a novel adipose derived hormone, is the factor which causes insulin resistance in animals. The recent data shows that resistin is also synthesized in human placenta. Hence, it is assumed that by modulation of cells sensitivity to insulin, resistin may contribute to development of diabetes during pregnancy. PMID- 16363388 TI - The surgeon general: why not have one? PMID- 16363387 TI - [Immunosuppressive drug therapy during pregnancy after kidney transplantation]. AB - The authors expounded present state of knowledge concerning immunosuppressive drugs therapy during pregnancy after kidney transplantation. Pregnancy is uncommon in women with end-stage renal disease treated with dialysis and in most cases it ends with pregnancy failure. Resuming the normal function of the ovaries after kidney transplantation substantially increases the chances of conception and successful pregnancy. The immunosuppression scheme and dosage of drugs used in pregnant women are vital to both the normal course of pregnancy and delivery of a healthy child. Considering the safety of the fetus it is acceptable to use prednisone, azathioprine, cyclosporine and tacrolimus. Due to the necessity to administer immunosuppressive drugs in relatively small doses, an important factor conditioning the normal course of pregnancy is maintaining a 1- or 2-year interval between the kidney transplantation and the conception. PMID- 16363389 TI - A consumptive finds salvation in nature. PMID- 16363390 TI - Neurologic consequences of cardiac arrest and preventive strategies. PMID- 16363391 TI - Doctors as athletes--review & commentary. PMID- 16363392 TI - A historical perspective on upper endoscopy at Rhode Island Hospital. PMID- 16363393 TI - Bio-emergency preparedness in Rhode Island: a role for urgent care centers? PMID- 16363394 TI - Creatine kinase: a review of its use in the diagnosis of muscle disease. PMID- 16363395 TI - Case presentations of the Brown Universty Department of Medicine Miriam Hospital Morbidity and Mortality Conference: a case of cardiac sarcoidosis. PMID- 16363396 TI - The Rhode Island HEALTH Web Query System. PMID- 16363397 TI - A physician's lexicon: the many words of diabetes mellitus. PMID- 16363398 TI - Is aging crummy deal? PMID- 16363399 TI - When memory fails: helping dementia patients remember God. PMID- 16363400 TI - Building a new culture of aging: revolutionizing long-term care. PMID- 16363401 TI - Mission Tsuanami relief: an eye-opening experience. PMID- 16363402 TI - Parish nursing meeting spiritual needs of elders near the end of life. PMID- 16363403 TI - Exploring advanced directives. PMID- 16363404 TI - The greatest of these is love. PMID- 16363405 TI - How can we help elders finish strong? PMID- 16363406 TI - Georgia hospitals offer hope and healing in tragedy. The state's health system reaches out to victims of Katrina. PMID- 16363407 TI - The power of the blue and white "H". PMID- 16363409 TI - A story you probably haven't heard. PMID- 16363408 TI - Finally: proposed rules for e-prescribing and EHR. PMID- 16363410 TI - Principles of peritoneal dialysis. PMID- 16363411 TI - Perceptions and experiences of pre-dialysis patients. AB - The literature shows that many complications including psychosocial, sexual and economic are associated with dialysis. However few studies have been conducted from the patients' perspective or have examined the pre-dialysis phase of established renal failure. The aim of this qualitative study was to explore the perceptions and experiences of pre-dialysis patients. Ten patients were interviewed, on one occasion, prior to starting dialysis. The interviews were conducted in a semi-structured manner in accordance with pre-defined interview topics. The findings of the study highlighted that participants had no clear expectations of dialysis treatment and generally expressed a sense of fatalism and lack of knowledge. This was highlighted by the main emergent theme, response to information. It became clear that most of the patients had received information, but had either misinterpreted, or been unable to absorb it. A number of specific issues including provision, quantity, interpretation and the timing of information appeared to be linked to the participants' inability to gain knowledge. These study findings highlight the importance of a patient centred, flexible education programme to enable patients to absorb and accurately interpret information. PMID- 16363412 TI - Simple pressure measurement is not reliable in detection of access stenosis in native AV fistulas. AB - Dynamic arterial and venous pressures (PA, PV) are used as the simplest tools to assess vascular access quality (VAQ). An increased PV over three consecutive dialyses is believed to indicate a stenosis, a rule devised for synthetic grafts (AVG) but not adequately validated for AV fistulas (AVF). In this study dynamic PV and static intra-access pressure (calculated by means of the simplified formula PIA=(PA+PV)/2) changes were evaluated in 46 accesses in which balloon angioplasty had to eventually be performed. The whole group consisted of 30 forearm AVF, 5 upper arm AVF and 11 AVG. Pressures were compared in each patient at a time of satisfactory access flow (QVA) and immediately before the angioplasty and pressure difference over that period (deltaPV, deltaPIA) evaluated. Despite a significant drop in QVA over the follow-up interval in both AVF and AVG, the mean deltaPV and deltaPIA in AVF were only several mm Hg and the chosen threshold limit of 20 mmHg was exceeded in approximately 10% of patients only. The results in the AVG group were, however, very different: The mean deltaPV and deltaPIA were close to 20 mmHg and almost 60% of patients in the AVG group exceeded this limit. Evaluation of PIA did not improve stenosis detection in either group. It is concluded that PV and/or PIA monitoring may be useful to detect a stenosis in AVG but not in AVF. PMID- 16363413 TI - On-line clearance monitoring for blood access management. AB - On-line Clearance Monitoring (OCM) provides frequent and precise information about urea clearance values during haemodialysis. In the case of blood access recirculation, it is presumed that urea clearance values on OCM would be lower and suspect to blood access malfunction. In order to check the relation between significantly lower urea clearance values and blood access recirculation, the Kt value (Clearance x time/min) for fifteen patients on OCM, including the patients with a low Kt/V in spite of their small urea distribution volume (V) was observed. Average urea clearance was calculated indirectly using Kt value (Kt/time in minutes = average clearance ml/min) and blood access recirculation tests performed using slow/stop flow two-needle, three samples method (urea method). After comparison of the recirculation percentage to clearance value, positive correlation between high recirculation and clearance reduction was noted. OCM alongside detection of haemodialysis inefficiency is also a practical instrument for blood access management between regular monitoring. Lower OCM urea clearance values demonstrate a possible blood access problem that can be confirmed with another method. When an OCM urea clearance reading is decreased by more than 25%, undiscovered access recirculation can be suspected. PMID- 16363414 TI - Anaemia as an early predictor for diabetic renal disease? A review of the literature. AB - Diabetes is the leading cause of end-stage renal disease in the western world and significant proportions of patients with diabetes mellitus develop renal complications. Anaemia is a key indicator of renal disease, and potentially may contribute to the pathogenesis of diabetes complications. Early correction of anaemia therefore has the potential to prevent or reverse other complications This review of the literature will examine anaemia in diabetic patients and seek to determine whether it is exclusive to diabetic nephropathy. PMID- 16363415 TI - A haemodialysis nutritional screening tool for nurses--a pilot study. AB - The purpose of the study was to pilot a nurse-performed nutritional screening tool (NST) for dialysis patients in order to identify nutritionally at-risk patients. Haemodialysis (HD) patients are at risk of nutritional-related problems. Nutritional screening by nurses may assist in the early recognition of and response to these problems. An NST was developed using 9 screening parameters. (BMI, weight change, poor appetite, GI symptoms, albumin, pre dialysis urea, K+, PO4++, HbA1c). The NST was compared with Standard Dietitian Assessment (SDA). 44 HD patients were screened with the NST and then with SDA. The tool showed sensitivity of 0.7 (95%CI+/- 0.21) and a specificity of 0.77 (95%CI+/-0.16). Reliability was low (alpha = .18). Alpha increased to 0.32 if pre dialysis urea was removed from the tool and increased to 0.48 if weight loss, appetite, K+ and PO4++ were used alone. The pilot study showed a low reliability of the NST compared with SDA. With further analysis and modifications, the NST has the potential to assist nutritional screening by nurses in dialysis centres that have limited dietetic access. PMID- 16363416 TI - Implementing nurse prescribing within the haemodialysis unit. AB - Patients within satellite haemodialysis units do not always have access to a medical practitioner. This may cause problems when prescriptions are required. Amendments to UK law to allow the introduction of supplementary prescribing came into force in 2003 allowing nurses with the appropriate experience, training and qualification to prescribe for their patients. Within a 14-station satellite unit a prescribing partnership has been successfully established. Clinical Management Plans have been implemented for haemodialysis patients. These include areas such as dialysis adequacy, access management, anti-coagulation, anaemia management, MRSA treatment and prophylaxis, antihypertensive therapy, calcium and phosphate control and exit site or line infection. 100% of patients within the unit have consented to the nurse prescribing for them under the Clinical Management Plan. Supplementary prescribing enhances nursing practice by empowering those who are best placed to make decisions regarding care and treatment for their patients. Patients appear confident in the ability of the nurse within the haemodialysis unit to prescribe competently. Nurse prescribing is of benefit to patient care, meeting the demands of an expanding patient population. It is recognition of the skill and experience required of haemodialysis nurses. PMID- 16363417 TI - Compliance in hypertension. AB - Despite improvements in the management of hypertension in the past several years, nearly 70% of patients with hypertension are not adequately controlled. Today's physician treating hypertension is armed with numerous efficacious antihypertensive agents. Thus, one of the major contributors to the large number of uncontrolled hypertensive patients appears to be non-compliance with prescribed regimens. Non-compliance is a universal characteristic and can affect all patients. However, the major problem of non-compliance is that it remains largely unrecognised in clinical practice. Several types of non-compliance exist with the 'drug holiday' being the most common. Good communication with the patient and the entire health care team is critical in increasing patient compliance. Long-acting medications that provide good blood pressure control beyond the 24-hour dosing period should be considered drugs of choice in non compliant hypertensive patients. This article will discuss different types of compliance, methods to measure compliance, and clinical consequences of non compliance in hypertension. PMID- 16363418 TI - A review of the updated European Best Practice Guidelines for the Management of Anaemia in Patients with Chronic Renal Failure. AB - The management of anaemia of Chronic Kidney Disease (CKD) has progressed significantly during the last decade. Research studies show that effective treatment of anaemia in adult patients with CKD can reduce cardiovascular complications slow progression of renal failure, and improve quality of life. To ensure adult patients receive optimum care, evidence-based practice and recognised guidelines and management strategies are needed to help reduce variations in clinical practice. In 1999 a working party of European Nephrologists developed the European Best Practice Guidelines (EBPG) for Management of Anaemia in Patients with Chronic Renal Failure to address this issue. Since the publication of the guidelines in 1999, over 3,000 papers relating to anaemia in CKD, have been published, many of which have significant implications for the practice of anaemia management. To ensure that the most up to date scientific evidence is available to those who manage anaemia in CKD the guidelines were revised in 2004. These guidelines, look at the anaemia management of adults with CKD and do not include guidelines for children. PMID- 16363419 TI - Evaluating staff and student experiences of multidisciplinary continuous professional development via distance-learning. AB - Continuous professional development (CPD) in caring for people with kidney disease is limited in some regions of the UK and within Europe generally. This is compounded for all by limited resources for course fees and the lack of study leave granted away from the clinical area for full-time courses. This is set against recommendations from National and European governments, and renal clinical guidelines concerning expectations of CPD and clinical competency levels of renal nurses (1-4). In the past renal practitioners have been trained in all areas of the renal speciality by Schools of Nursing linked to renal units based in large teaching hospitals. However, more recent changes in the structure of Health Care provision have led in some instances to a rationalising of post registration education delivery. PMID- 16363421 TI - [Reducing the proportion of patients with low physical activity]. PMID- 16363422 TI - [An outbreak of lymphogranuloma venereum in Western Europe]. AB - Lymphogranuloma venereum (LGV) is a sexually transmitted infection caused by Chlamydia trachomatis serotypes L1, L2, and L3. It is endemic in tropical areas, while seldom in industrialised countries. Since 2002, an outbreak of rectal LGV has been observed among men having sex with men, in Netherlands, France, and other countries of Western Europe. After an incubation period of 2 days to 2 months, LGV develops in three stages: ulcerated papule; inflammatory satellite lymph node and/or proctitis; chronic inflammatory complications (fistulas and sclerosis). Chlamydia trachomatis infection is confirmed by PCR and serovars is identified by RFLP of omp1 gene amplified by PCR. Recommended first line treatment is doxycycline for at least 21 days. PMID- 16363423 TI - [Pulmonary tuberculoma]. PMID- 16363424 TI - [Cardiovascular risk factors: on which elements to base the actions of prevention?]. PMID- 16363425 TI - [Cardiovascular risk factors: an update]. AB - Cardiology is the field of medicine where therapeutic progress has been the most outstanding during the last decades. But this progress can only be maintained if two conditions are fulfilled: firstly, patients should have time to access to acute care, secondly, if they survive, they should not recur. Today, this is possible because of the progress of primary and secondary prevention. Knowledge of risk factors is important and efficiency of therapeutic and preventive strategies has been demonstrated. As reported in recent studies, action against a limited set of classical risk factors would allow to reduce by half at least cardiovascular attack rates. To reach this goal, global cardiovascular risk should be considered. However, efficient results will only be obtained if these preventives measures are implemented as a deal between the physician and his patient, and if they last longer enough. PMID- 16363426 TI - [Metabolic syndrome and cardiovascular risk]. AB - Metabolic syndrome points out numerous vascular risk factors clustering together, due to a common mechanistic substratum, visceral obesity and insulin resistance. The definitions of the syndrome allow to identify individuals at risk of vascular disease and diabetes but actually do not add anything to the predictive value of the individual components of the syndrome. Metabolic syndrome could become a target for some specific therapeutic interventions. PMID- 16363427 TI - [Cardiovascular prevention and large clinical trials]. AB - Large clinical trials of the three last decades have transformed the therapeutic landscape in depth. After putting into question the multi-factorial approach and simplistic models such as that of isotropy, clinical trials have shown to which extent some drugs are an efficacious way of reducing the burden of premature cardiovascular events and of prolonging life. The optimal use of the results from large clinical trials requires a permanent updating process, which is not realistic for the non-specialist physician: dedicated and fitted interfacing tools have to be developed. Practitioners must be trained to use these new concepts and tools. PMID- 16363428 TI - [Cardiovascular prevention of high risk patients by hypolipidemic treatment]. AB - Afssaps guidelines 2005 for the treatment of dyslipidemic patients provide a valuable tool for optimal treatment in clear cut situations. However, for patients with intermediate cardiovascular risk or when it might be necessary to intensify hypolipidemic treatment, additional risk markers can be contributive for risk estimation (CRP, intima-media thickness) and it is usefull to take into account the physiopathology of dyslipidemia and subgroup analysis of clinical trials. An efficient cardiovascular prevention implies a screening of high cardiovascular risk subjects, an optimal treatment and conversely not to over treat low risk subjects. In France, convergent studies show under treatment of high risk subjects and over treatment of low risk dyslipidemic subjects. PMID- 16363429 TI - [AFSSAPS guideline for the treatment of dyslipidemia]. PMID- 16363430 TI - [Lifestyle and cardiovascular risk]. AB - This paper describes the relation between lifestyle and cardiovascular risk globally and for each lifestyle risk factor. In 3 large studies (Interheart, HALE project and the Nurses' health study), four risk factors have been identified: smoking, diet, low level of exercise and low or no alcohol consumption. Prevention of atherosclerotic diseases can be obtained by smoking cessation, diet modifications aimed at lowering body weight and having a Mediterranean style diet, continuing lifelong physical activity such as walking more than two hours a week. Alcohol consumption, although related to lower risk, cannot be prescribed as a preventive action. PMID- 16363431 TI - [Choosing the right medication for the right patient]. AB - Over the past 15 years, the efficacy of several classes of medications on the prevention and treatment of atherosclerosis has been documented. The objectives of medical therapy are triple: to avoid or control the evolution of the atherosclerotic process, control associated risk factors, and, whenever appropriate, to control symptoms. Four classes of medications have documented a major clinical benefit by decreasing mortality and severe adverse cardiovascular events: antiplatelet agents, beta-blockers, statins and ACE inhibitors. These medications should therefore constitute first line therapy. Beside the use of medications, life style changes (Mediterranean diet, physical exercise, smoking cessation) are also necessary. PMID- 16363432 TI - [Searching cardiovascular risk factors in MEDLINE]. PMID- 16363433 TI - [The health importance of physical activity]. PMID- 16363434 TI - [Health literacy and comorbidity. Reading is fundamental]. PMID- 16363435 TI - [National ranking exam. N10. What could happen at the exam?]. PMID- 16363436 TI - [Sudden vision loss]. PMID- 16363437 TI - [Chronic renal failure]. PMID- 16363438 TI - [Gastric tumours]. PMID- 16363439 TI - [Acute pancreatitis]. PMID- 16363440 TI - [A great and forgotten physician: Galen of Pergamon]. PMID- 16363441 TI - Occupational health in the construction industry. PMID- 16363442 TI - Mortality and morbidity among bridge and tunnel construction workers who worked long hours and long days constructing the Great Belt Fixed Link. AB - OBJECTIVES: This study aimed at estimating the mortality and morbidity of hospital treatment among bridge and tunnel workers who worked round the clock, long hours, and long weeks to construct the Great Belt Fixed Link. METHODS: A cohort of all Danish workers in the construction industry was followed for death and first hospitalization and outpatient or emergency ward treatment over 6 years. Standardized mortality and morbidity ratios (SMR) were calculated for selected diagnoses, and 5123 bridge and tunnel construction workers were compared with all 109 383 Danish construction workers. RESULTS: The comparison showed an overall SMR of 124 with a 95% confidence interval (95% CI) of 97-155. For infectious diseases and intestinal infectious diseases, the corresponding values were 156 (95% CI 132-184) and 167 (95% CI 117-230), respectively. For diseases of the nervous system, it was 138 (95% CI 118-160), and including nerve, nerve root and plexus disorders it was 135 (95% CI 104-171), for instance, mononeuropathies of the upper limbs (SMR 136, 95% CI 101-180). The SMR was 139 (95% CI 126-153) for circulatory diseases, 157 (95% CI 130-189) for ischemic heart disease, 129 (95% CI 114-146) for diseases of the respiratory system, 124 (95% CI 114-135) for diseases of the digestive system, and 115 (95% CI 108-123) for diseases of the musculoskeletal system and connective tissue, including other intervertebral disc disorders (than cervical) (SMR 130, 95% CI 109-154). CONCLUSIONS: Bridge and tunnel workers who work round the clock, long hours, and long weeks have a mortality rate that is as high as that of other construction workers, and they are treated more often in hospitals. PMID- 16363443 TI - Asbestos hazard in the Swedish construction industry--recent trends in mesothelioma incidence. AB - OBJECTIVES: The aim of this study was to analyze the incidence of pleural tumors among various categories of Swedish construction workers and to determine to what extent its change over time differs from that of the general male population. METHODS: Traditional methods have been used to study cancer incidence through 1998 in a cohort comprising 370 165 male workers examined in 1971-1992 by Bygghalsan, an organization at the time providing nationwide occupational health service. Incidence was assessed by linkage to the national cancer register. RESULTS: Swedish construction workers, particularly those heavily exposed to asbestos, had an excess incidence of pleural tumors in 1975-1998 [standardized incidence ratio (SIR) 3.16, 95% confidence interval (95% CI) 2.55-3.88]. The excess declined with subsequent follow-up periods and birth cohorts with the exception of the most recent period (SIR 3.83, 95% CI 2.64-5.38) and those borne in the 1930s. CONCLUSIONS: A possible decline in pleural tumors among men following the cessation of asbestos use 25 years earlier in the population at large may not be applicable to an end-user sector like construction work. In occupations charged with repairing and refurbishing work, there may even have been an increase lately. PMID- 16363444 TI - Development of a decision model to identify workers at risk of long-term disability in the construction industry. AB - OBJECTIVE: This study presents a decision model that predicts long-term disability among construction workers. METHODS: Risk factors were identified in two cohort studies among construction workers and evaluated in validation samples of smaller cohort studies among Dutch construction workers. The risk estimates (odds ratios) were used in a logistic regression model to calculate the probability of long-term disability in the next 4 years for a particular construction worker, subject to a specific combination of risk factors. The a priori probability was set equal to the overall long-term disability risk among the youngest construction workers (< 30 years) with a relatively short exposure history. RESULTS: According to literature findings, the risk estimate for work ability was set with the odds ratio at 2.0 for good work ability, 5.0 for moderate work ability, and 10.0 for bad work ability. Age-dependent risks were set at odds ratios of 1.5, 2.0, and 3.0 for the age groups of 30-34 years, 35-44 years, and 45-54 years, respectively. A sickness absence period of > or = 3 months had an odds ratio of 2.0, and severe musculoskeletal complaints had an odds ratio of 3.0. Since the number of construction workers older than 55 years was rather small and heavily biased by a healthy worker effect, it was decided to limit the applicability of the decision model to workers aged 20-55 years. The decision model used four risk factors and predicted a 40-fold difference in disability risk between construction workers with all four risk factors present (0.79) and those without any risk factor (0.02). CONCLUSIONS: The decision model presented the combined effect of different risk factors on the risk of an individual worker becoming disabled within 4 years. Evaluation studies will need to demonstrate whether the application of this decision model is helpful in identifying workers at risk for long-term disability and will facilitate appropriate intervention at the individual level. PMID- 16363445 TI - The conditions and perception of work during painting behind screening. AB - OBJECTIVE: This study attempted to reveal whether airborne solvent concentrations during painting behind screening are comparable to solvent concentrations during painting indoors, what factors potentially influence work conditions during painting behind screens, both positively and negatively, and what can be done to eliminate the disadvantages of screening. METHODS: Measurements of volatile organic compounds (VOC), climate, light, and noise were made at three sites during 1 day each. VOC were measured at another 10 sites, again during 1 day each. Questionnaires were sent to painters working in 305 firms. RESULTS: The results of the first series of measurements (exposure to VOC) ranged from 3.4 to 22.9 mg/m3 with a geometric mean of 11.7 and a geometric standard deviation of 2.1. This range was less than the level expected in well-ventilated rooms. In the second series of measurements, with a few exceptions, the concentrations of the individual compounds were below 1 mg/m3. In both series, the exposure index was generally < or = 0.05. The advantages of working behind screening were more certainty of work during the winter months, better temperature conditions, better quality of work, less draft, and less temperature fluctuation. CONCLUSIONS: Painting behind screening, as studied in this project, results in low exposure to VOC when compared with indoor situations. The main factors that need to be improved are temperature, lighting, view from the workplace, and ventilation. PMID- 16363446 TI - Quantitative risk assessment in relation to occupational exposure to polychlorinated biphenyls in the removal of old sealants from buildings. AB - OBJECTIVES: The objectives of the study were to assess exposure to polychlorinated biphenyls (PCB) among workers involved in the removal of old sealants containing PCB and to evaluate the related long-term health risks by means of quantitative risk assessment. METHODS: Workers' exposure via different exposure routes was estimated from air and material samples and exposure calculations. The health risks were evaluated using unit risks and a reference dose based on animal tests and evaluations made by the Environmental Protection Agency in the United States. RESULTS: The estimated exposure of the workers was higher than that of the general population. It was about 10-fold higher than the reference dose and average dietary intake (both 0.02 microg/kg-day). The calculated point estimate of excess cancer risk was 4.6 x 10(-4) cancer cases per lifetime. CONCLUSIONS: The estimated excess cancer risk among workers was low, although the PCB exposure of these workers was higher than the dietary intake of the general population. This quantitative risk assessment method can be used to evaluate the health risk of groups of workers by occupational health professionals. PMID- 16363447 TI - Risk assessment of silicosis and lung cancer among construction workers exposed to respirable quartz. AB - OBJECTIVES: The aim of this study was to assess the magnitude of the silicosis and cancer risk among construction workers. METHODS: In 1998, 1335 of 4173 invited construction workers with expected high cumulative exposure to quartz were studied for early signs of silicosis. In 2002 the study was repeated for 96 persons. Exposure measurements were performed among 34 construction workers. Silicosis risk was assessed by converting study results to the whole group of construction workers and by risk analysis based on exposure data combined with documented exposure response relations. Excess risk for cancer was also calculated from available exposure measures. RESULTS: The initial study among construction workers revealed a prevalence of 0.8% of workers with rounded opacities on chest X-rays. The follow-up showed a much higher percentage (12%) of persons with rounded opacities on X-rays. The results were confirmed by high resolution computed tomography. It was estimated that roughly 9% of the population initially studied (N = 1335) would have been observed with rounded opacities at follow-up. On the basis of the exposure data, a lifetime risk of silicosis above 5% is expected for workers exposed to levels above the occupational exposure limits. An excess lifetime risk for lung cancer is expected when workers are exposed to quartz levels above the occupational exposure limit. Due to the scarcity of exposure data, an estimation of the size of the group at risk is not yet possible. CONCLUSIONS: All available data indicate that construction workers exposed to quartz levels above occupational exposure limits are clearly at elevated risk of silicosis and other respiratory diseases. PMID- 16363448 TI - Dose-response associations between musculoskeletal disorders and physical and psychosocial factors among construction workers. AB - BACKGROUND: As musculoskeletal disorders are common in general populations and they therefore constitute a heavy burden to society, the identification of their determinants is important. The aim of this study was to identify dose-response associations for location-specific prevalences of musculoskeletal disorders with various physical exposures and psychosocial factors in a large group of workers. METHODS: The study was based on cross-sectional data from self-administered questionnaires filled out by 85 191 male employees in the Swedish construction industry. In addition to symptoms from nine musculoskeletal locations, the questionnaire addresses physical exposures and psychosocial factors. Logistic regression methods were used to study disorder prevalence in relation to scales of these exposures and factors. RESULTS: Compared with working rarely in a stooping or twisted posture, the odds ratio (OR) for lower back disorder with respect to often working in this posture was 3.05 [95% confidence interval (95% CI) 2.73-3.42]. The OR values for work with hands above the shoulders and for neck and shoulder disorders were 2.93 (95% CI 2.65-3.24) and 3.66 (95% CI 3.32 4.04), respectively. Among the psychosocial factors, sleeping problems and hurrying without reason were associated with symptoms in all of the body locations, the highest OR being found for neck symptoms and sleeping problems, 2.44 (95% CI 2.15-2.78). CONCLUSIONS: Work in awkward postures has strong location-specific dose-response associations. In contrast, some psychosocial factors, primarily those reflecting characteristics of individual persons, are strongly associated with symptoms in all body locations. Symptoms seem not to be associated with factors like support from workmates or supervisors or control of the work situation. All location-specific prevalence rates seem to increase strongly with age. PMID- 16363449 TI - Knee-straining work activities, self-reported knee disorders and radiographically determined knee osteoarthritis. AB - OBJECTIVES: Earlier studies indicate an increased risk of knee disorders in trades with knee-straining work positions, but no dose-response correlation has yet been documented. This study examined whether self-reported knee complaints and radiologically diagnosed knee osteoarthritis are dose-related to kneeling and squatting work positions. METHODS: Worktasks for floor layers and carpenters were video-recorded, and the time spent in knee-straining work positions was quantified. A questionnaire study included 133 floor layers, 506 carpenters, and 327 compositors, all without earlier acute knee trauma. Radiological examinations were carried out on a stratified sample (N = 150) from the questionnaire study. The individual exposure was calculated from the amount of knee strain quantified in the video-recorded worktasks, the self-reported time spent in the worktask, and the number of years in the trade. Statistical analyses were carried out with a binary logistic regression model, adjusting for age, body mass index, smoking, and knee-straining sports activities. RESULTS: The odds ratios for self-reported knee complaints and radiographically determined knee osteoarthritis were increased in relation to the degree of knee-straining work demands for workers with low-to-moderate, high, and very high exposure to knee-straining work demands in a comparison with the reference group, without knee-stressing work activities. CONCLUSIONS: The results indicate that there is a dose-response correlation between knee-straining work activities and the development of self-reported knee complaints and radiologically diagnosed knee osteoarthritis. PMID- 16363450 TI - Effectiveness of measures and implementation strategies in reducing physical work demands due to manual handling at work. AB - This review aimed at producing insight into the effectiveness of interventions to reduce the physical work demands associated with manual (materials) handling in the work situation and musculoskeletal symptoms in the longer-term. A systematic electronic literature search between 1990 and February 2003 was performed in the following databases: Embase, Medline, HSE-line, Nioshtic, and Nioshtic-2. Three inclusion criteria were applied, and altogether 44 studies were included for review. The interventions were divided into two categories, (ergonomic) measures and implementation strategies. One randomized controlled field study was found that established a causal effect for a combination of implementation strategies in reducing the physical work demands associated with manual handling and reducing acute musculoskeletal symptoms. All four of the controlled field studies showed a significant reduction in the physical work demands when lifting devices were part of the intervention. Two of these studies measured a significant reduction in low-back disorders in the longer term. Several uncontrolled pre-post studies showed effects in the same direction. Of the 26 implementation strategies, 21 that measured an improvement in the process variables (eg, aimed behavioral variables) used a participatory ergonomics approach, an education (or training) program or both with the direct involvement of workers. It was concluded that significant reductions in physical work demands and musculoskeletal symptoms were found when (mechanical) lifting devices were part of the intervention. The higher quality studies that showed improvement in behavior indicate the importance of using facilitating and educational strategies in the implementation of ergonomic measures. PMID- 16363451 TI - Physical load exposure at construction sites. AB - OBJECTIVES: This field study analyzed work-related causes of musculoskeletal disorders to investigate the structure of physical workload in different occupations in the construction industry and rank different tasks with respect to the load on the lumber spine. METHODS: An observation instrument (Arbeitswissenschaftliches Erhebungsverfahren fur Bauarbeiten) and a specially devised data retrieval system (Allgemeines Datenerfassungs- und Analysesystem fur Bauarbeit) provided material for a large database which allowed a differentiated analysis of load exposures. The study was comprised of data from 340 construction workers (bricklayers, scaffolders, carpenters, plumbers, and painters). On the basis of a regular daily worktime of 8 hours, specific statistical aspects were studied concerning manual materials handling, biomechanical pressure on lumbar disc L5/S1, and posture constraints during kneeling, squatting, bending and overhead positions. RESULTS: The scaffolders (13.7% of the regular daily worktime), bricklayers using bricks requiring two hands (7.1%), and carpenters (6.7%) handled weights of > 10 kg. With respect to lumbar disc L5/S1, the scaffolders and bricklayers often showed pressures in excess of 3.4 kN. Bricklaying required bent postures for 20.7-35.6% of the daily worktime. The painters (23.8%), plumbers (16.7%), and carpenters (7.2%) often worked in kneeling postures. The painters often used overhead positions (18.3%). The bricklayers and scaffolders had high frequencies of materials handling. The recovery time for this repetitive work was longer than threefold the load time. CONCLUSIONS: This study showed that it is possible to rank different construction tasks with respect to load exposure. In addition it was shown that preventive measures such as improved ergonomic design, organizational structure, training, and medical health care are needed. PMID- 16363452 TI - Conceptual framework for the implementation of interventions in the construction industry. AB - OBJECTIVES: The objective of this study was to explore the necessary steps to define the implementation of interventions aimed at reducing physical work demands due to manual materials handling in the construction industry. METHODS: A theoretical structured framework of six steps is outlined as a method for developing the implementation of interventions. In this framework, both the proposal for implementing the intervention measures and the context analysis are conditional. RESULTS: Application of the framework in the development of the implementation of interventions to reduce the physical work demands for bricklayers and bricklayers' assistants has been presented. CONCLUSIONS: The framework was found appropriate for defining implementation strategies and (ergonomic) measures in the construction industry. PMID- 16363453 TI - Efficiency in reducing lost-time injuries of a nurse-based and a first-aid-based on-site medical facility. AB - OBJECTIVES: The purpose of the study was to evaluate the efficiency of two standards for on-site medical facilities in reducing lost-time injuries during the construction of the link across the Oresund sound between Sweden and Denmark. One medical facility employed licensed nurses, who had advanced medical assistance at their disposal. The other medical facility utilized first-aid trained watchmen. The on-site medical facilities aimed both at providing immediate medical assistance to workers subjected to occupational injuries and at reducing lost worktime. METHODS: The distributions of injuries treated on-site (the worker resumed work after treatment) and injuries sent to hospitals or to specialists were compared for each type of injury and for each category of injured body part. RESULTS: The on-site medical facilities dealt, in particular, with the treatment of ocular injuries (21%), wounds (21%), and sprains or strains (15%). The study showed a statistically significant on-site treatment (and resume work) rate ratio of 3.3 between the nurse-based (76%) and the first-aid-based (23%) medical facility. CONCLUSIONS: The construction of the Oresund Link shows a need for on-site medical facilities, particularly at remote construction sites, and that it is essential that the medical personnel have both the qualifications and authorization to treat site-specific workplace injuries effectively in order to obtain high on-site treatment rates. PMID- 16363454 TI - The Galaxen model--a concept for rehabilitation and prevention in the construction industry. AB - The Galaxen model was developed during the late 1980s to provide rehabilitation and prevention activities in the construction industry. It handles around 1200 workers with long-term sick leave or partial disabilities annually, some 10% of whom annually leave Galaxen for an ordinary job without a wage subsidy. The model includes a decision by a rehabilitation board of representatives from the employers, the trade union, and the regional employment office, a rehabilitation plan, allotment of a case manager, wage subsidies from the State to the company, a search for a suitable job in relation to the partial disability. It also includes a preventive program with emphasis on practical ergonomics. The Galaxen model has proved to be a suitable means of rehabilitating construction workers and returning them to the workforce. The model was developed within the Swedish social security system but could well be adjusted to other contexts. PMID- 16363455 TI - My journal years, 1972 to 1977--a look back. PMID- 16363456 TI - A memoir of a sojourn as editor of The Canadian Journal of Psychiatry. PMID- 16363457 TI - Editing the Canadian Journal of Psychiatry 1995-2004. PMID- 16363458 TI - Do psychiatric journals have a future in the age of the Internet? PMID- 16363459 TI - Screening for complicated grief: when less may provide more. AB - OBJECTIVE: This study aimed to discover screening questions for initial assessments of psychiatric outpatients to identify which patients are likely experiencing complicated grief (CG). METHOD: We examined the responses of 235 outpatients to questionnaire items. One set came from the Texas Revised Inventory of Grief (TRIG). This set represented events cited in clinical literature as indicators or risk factors for CG. Another set came from a factor analysis of items that define CG. We determined relations between items and CG, analyzed sensitivity and specificity, and compared the 2 sets of items. RESULTS: The factor analysis items were superior. The 2 best items correctly identified nearly 90% of patients with and without CG. CONCLUSION: The items can be transformed into screening questions to be used in initial interviews. PMID- 16363460 TI - Child abuse, psychiatric disorder, and running away in a community sample of women. AB - OBJECTIVE: Studies involving clinical samples of adolescents indicate that running away is a common problem associated with substantial emotional impairment, yet little information is available from community-based samples. This study aimed to investigate runaway behaviour in female adolescents with and without a history of maltreatment in childhood. METHOD: We analyzed data from a province-wide community survey conducted in Ontario (n = 3760 girls and women, aged 15 to 64 years). Respondents returned a self-completed questionnaire regarding their history of maltreatment as children. Psychiatric disorder was assessed with a revised version of the Composite International Diagnostic Interview. RESULTS: Respondents who reported physical abuse only, sexual abuse only, and physical abuse with sexual abuse were 2 to 4 times more likely to report running away from home before age 16 years. Parental psychiatric disorder, respondent psychiatric disorder, respondent age, and income were also significant correlates of running away. CONCLUSIONS: Understanding the relation between exposure to maltreatment as a child, psychiatric disorder, and running away in women is important for determining ways to prevent a behaviour that puts youth at high risk for a wide range of difficulties. PMID- 16363461 TI - Completed suicides in a youth centres population. AB - OBJECTIVE: From 1995 to 2000, 422 youths, aged 18 years and under, died as a result of suicide in Quebec. More than one-third had received services from youth centres (YCs) at some point. This study sought to characterize a sample of those youths to improve services for this at-risk population. METHOD: From a retrospective study of YCs and coroner's office files, we investigated the clinical features of youths who had received YC services and died by suicide. We compared them with YC patients matched for age, sex, and geographic area who had reported suicidal behaviour or who had no such symptoms. RESULTS: Among those who committed suicide, we found a ratio of 3.8 boys for 1 girl, with a mean age of 16.8 years. Hanging, used by 73.6%, was the most frequent means; 53.6% had a previous suicide attempt. The group that committed suicide had more indicators of major depression, substance abuse, and disruptive behaviours, as well as more adverse events. CONCLUSIONS: Interventions should focus on screening for mental disorders and suicidal behaviours on the initial contact with YC services. This screening should be implemented through a medical multidisciplinary team that includes psychoeducational services. PMID- 16363462 TI - Neuroactive steroids in schizophrenia. AB - Schizophrenia is a psychiatric disorder with a complicated pathophysiology, involving many biochemical abnormalities in the brain. Because neuroactive steroids (NASs) modulate neurotransmitter systems that are implicated in the pathology of schizophrenia, recent research has focused on examining the role that NASs play in the illness. Although research in this area is relatively new, it appears that NASs may potentially be implicated in the pathophysiology of the illness. This paper reviews the current understanding of NASs, the research literature on NASs in schizophrenia and in animal models of the illness (including the effects of antipsychotic medication on NASs) and on the potential antipsychotic role of NASs themselves and, finally, discusses future directions for this area of schizophrenia research. PMID- 16363463 TI - The role of mental health legislation. PMID- 16363464 TI - Tardive dyskinesia in the era of typical and atypical antipsychotics. Part 2: Incidence and management strategies in patients with schizophrenia. AB - OBJECTIVE: Tardive dyskinesia (TD), the principal adverse effect of long-term conventional antipsychotic treatment, can be debilitating and, in many cases, persistent. We sought to explore the incidence and management of TD in the era of atypical antipsychotics because it remains an important iatrogenic adverse effect. METHODS: We conducted a review of TD incidence and management literature from January 1, 1965, to January 31, 2004, using the terms tardive dyskinesia, management, therapy, neuroleptics, antipsychotics, clozapine, olanzapine, risperidone, quetiapine, ziprasidone, and aripiprazole. Additional articles were obtained by searching the bibliographies of relevant references. We considered articles that contributed to the current understanding of both the incidence of TD with atypical antipsychotics and management strategies for TD. RESULTS: The incidence of TD is significantly lower with atypical, compared with typical, antipsychotics, but cases of de novo TD have been identified. Evidence suggests that atypical antipsychotic therapy ameliorates long-standing TD. This paper outlines management strategies for TD in patients with schizophrenia. CONCLUSION: The literature supports the recommendation that atypical antipsychotics should be the first antipsychotics used in patients who have experienced TD as a result of treatment with conventional antipsychotic agents. The other management strategies discussed may prove useful in certain patients. PMID- 16363465 TI - [Professional integration of individuals with a mental illness: an analysis of individual characteristics]. AB - A prospective study of individuals with a mental illness aimed to identify significant determinants of their integration in the work force. Regression and mean comparison analysis allowed exploring an individual's most typical characteristics that could explain his or her socio-professional integration, while considering several professional integration indicators. First, results suggest that linked to work (e.g. commitment to work) and socio-demographic variables (e.g. financial support received) can predict performing a work activity and obtaining competitive employment. Second, 2 additional variables can predict autonomy in employment, which are a feeling of efficiency in looking for work and a capacity to overcome obstacles to work integration. Third, cognitive and clinical variables are associated with a delay to start work. Fourth, the number of hours worked per week is explained by the feeling of efficiency in overcoming health problems and others motivational and psychosocial variables. Finally, it is important to clearly identify the professional integration indicators in order to determine the significant predictors. PMID- 16363466 TI - Re: The prevalence of psychological morbidity in West Bank Palestinian children. PMID- 16363467 TI - Forensic risk assessment and dangerous driving. PMID- 16363468 TI - Separation methods for toxic components in Traditional Chinese Medicines. AB - Traditional Chinese medicines (TCMs) with many unique functions for treating diseases have attracted the interest of people worldwide. They have been popularly utilized for therapy and health promotion in most Asian countries and even in many European and North American countries. However, it should be clearly noted that TCMs are mixtures with complicated composition usually containing hundreds, even thousands of chemically different constituents, and it is the multiple constituents that work synthetically to determine the ultimate effect of a formula of TCM. Meanwhile, some components with toxicity in some TCMs, having various negative effects on different parts of body, may do serious harm to people's health; such harm in particular requires our attention. In this article, applications of different chromatographic and electrophoretic techniques in the analysis of toxic components in TCMs in recent decades have been comprehensively reviewed and some hyphenated procedures (combinations of two kinds of measurement) applied in this field are also summarized. PMID- 16363469 TI - Determination of total gaseous lead in the atmosphere by honeycomb denuder/electrothermal atomic absorption spectrometry. AB - A technique is described for the determination of total gaseous lead in the atmosphere by honeycomb denuder collection, followed by an electrothermal atomic absorption spectrometry (ETAAS) measurement. The collection efficiency of the honeycomb denuder in which a solution containing 2% HNO3/2% glycerine/1% ammonium dihydrogenphosphate was coated for trapping the gaseous lead in the atmosphere was 98.8%. The linear absorbance response was obtained for a concentration range of 0-1.39 microg m(-3) of lead in the atmosphere. A precision of 4.8% RSD (peak height absorbance, n = 11) for an aqueous solution of 1 ng of lead standard, characteristic masses (CM) of 23 pg and detection limit (3sigma) of 54 pg for an aqueous solution of 0.01 ng lead standard was achieved with 100 microg ammonium dihydrogenphosphate as a chemical modifier. The average recovery of lead in three standard samples prepared by the independent digestion of NIST SRM 1648 (Urban Particulate Matter) using our analytical system was 97.8%. The total content of the gaseous lead in the atmosphere of our laboratories was 0.35-0.38 microg m( 3). PMID- 16363470 TI - New conventional coated-wire ion-selective electrodes for flow-injection potentiometric determination of chlordiazepoxide. AB - New chlordiazepoxide hydrochloride (Ch-Cl) ion-selective electrodes (conventional type) based on ion associates, chlordiazepoxidium-phosphomolybdate (I) and chlordiazepoxidium-phosphotungstate (II), were prepared. The electrodes exhibited mean slopes of calibration graphs of 59.4 mV and 60.8 mV per decade of (Ch-Cl) concentration at 25 degrees C for electrodes (I) and (II), respectively. Both electrodes could be used within the concentration range 3.16 x 10(-6)-1 x 10(-2) M (Ch-Cl) within the pH range 2.0-4.5. The standard electrode potentials were determined at different temperatures and used to calculate the isothermal coefficients of the electrodes, which were 0.00139 and 0.00093 V degrees C(-1) for electrodes (I) and (II), respectively. The electrodes showed a very good selectivity for Ch-Cl with respect to the number of inorganic cations, amino acids and sugars. The electrodes were applied to the potentiometric determination of the chlordiazepoxide ion and its pharmaceutical preparation under batch and flow injection conditions. Also, chlordiazepoxide was determined by conductimetric titrations. Graphite, copper and silver coated wires were prepared and characterized as sensors for the drug under investigation. PMID- 16363471 TI - Cu(II)-3-(5-chlor-2-hydroxy-3-sulfophenylazo)-6-(2,4,6-tribromophenylazo)-4,5 dihydroxynaphthalene-2,7-disulfonic Acid-Co(II) binuclear complexation and its application to the selective determination of cobalt at ng/ml level. AB - The chromophore, 3-(5-chlor-2-hydroxy-3-sulfophenylazo)-6-(2,4,6 tribromophenylazo)-4,5-dihydroxynaphthalene-2,7-disulfonic acid (CSTDD) was used to complex Cu(II) and Co(II) in aqueous solution at pH 9.43. A binuclear complex of Cu-CSTDD-Co was formed and showed a high selectivity for the determination of Co(II). The spectral correction technique was applied to characterize the complexes. The results showed the formation of complexes of Cu(CSTDD), Co(CSTDD)3 and Cu2(CSTDD)2Co. The quantitative analysis of Co(II) at ng/ml level was carried out by the light-absorption ratio variation approach (LARVA). The results showed that the technique is satisfactory to determine Co(II) at trace level in water samples with a detection limit of 2.3 ng/ml. PMID- 16363472 TI - Fast simultaneous determination of niobium and tantalum by Kalman Filter analysis with flow injection chemiluminescence method. AB - A fast and highly efficient Kalman Filter analysis-flow injection chemiluminescence (FI-CL) method was developed to simultaneously determine trace amounts of niobium and tantalum in geological samples. The method, without the boring process of separation and dear instruments, is suitable for field scene analysis. The mixed chemiluminescence kinetic curve was analyzed by a Kalman Filter (KF) in this method to realize the simultaneous determination of niobium and tantalum. Possible interference elements in the determination were investigated. Under the selected conditions, the detection limits (3sigma, n = 11) of niobium(V) and tantalum(V) were 2.1 x 10(-3) microg g(-1) and 4.0 x 10(-3) microg g(-1), respectively, and the relative standard deviations were 4.9% and 3.3% (n = 9). The method was applied to the determination of niobium and tantalum in geological samples with satisfactory results. PMID- 16363473 TI - Study on degradation characteristics of chitosan by pepsin with piezoelectric quartz crystal impedance analysis technique. AB - Piezoelectric quartz crystal impedance analysis technique was applied to study the chitosanolytic activity of pepsin. The method is based on the viscosity density reduction of chitosan solution during the enzymatic degradation process. Experiments examined the time courses of the variations of motional resistance (deltaR1) for a quartz crystal. By comparing the deltaR1 response curves under different degradation conditions, the effects of pH, temperature, enzyme and substrate concentration on the chitosanolytic activity of pepsin was investigated in detail. The results suggest that the optimum pH and temperature were 4.6 and 55 degrees C, respectively. Increasing aptly the enzyme or substrate concentration was in favor of the degradation of chitosan. Moreover, the influence of the degree of deacetylation (DD) on the enzymatic degradation was studied. The result indicates that chitosan with a lower DD was easier to be degrade compared with chitosan with a higher DD. Also, it was found that there was a good linear relationship between the deltaR1 response and the DD value. The regression equation was deltaR1 = 0.058 x DD-6.795 and the correlation coefficient was 0.987. PMID- 16363474 TI - Flotation-separation and ICP-AES determination of ultra trace amounts of copper, cadmium, nickel and cobalt using 2-aminocyclopentene-1-dithiocarboxylic acid. AB - A rapid flotation method for separation and enrichment of ultra trace amounts of copper(II), cadmium(II), nickel(II) and cobalt(II) ions from water samples is established. At pH 6.5 and with sodium dodecylsulfate used as a foaming reagent, Cu2+, Cd2+, Ni2+ and Co2+ were separated simultaneously with 2-aminocyclopentene 1-dithiocarboxylic acid (ACDA) added to 1 l of aqueous solution. The proposed procedure of preconcentration is applied prior to the determination of these four analytes using inductivity coupled plasma-atomic emission spectrometry (ICP-AES). The effects of pH, concentration of ACDA, applicability of different surfactants and foreign ions on the separation efficiency were investigated. The preconcentration factor of the method is 1000 and the detection limits of copper(II), cadmium(II), nickel(II) and cobalt(II) ions are 0.078, 0.075, 0.072 and 0.080 ng ml(-1), respectively. PMID- 16363475 TI - Flow-injection determination of ascorbic acid and cysteine simultaneously with spectrofluorometric detection. AB - A simple and sensitive spectrofluorometric method was developed for the simultaneous determination of ascorbic acid and cysteine by a flow-injection system. This method is based on the reduction of Tl(III) with ascorbic acid or cysteine in acidic media, producing fluorescence reagent, TlCl3(2-) (lambdaex = 227 nm, lambdaem = 419 nm). The injected sample solution was divided into two separate streams. The first stream was treated with Tl(III) at pH 3.0 and then passed through a 270 cm reaction coil to the flow cell of the spectrofluorometer, where the fluorescence intensity was measured. This signal is related to ascorbic acid and cysteine concentration. The second part of the injected sample solution was treated with Tl(III) in HCl solution and then passed through a 50 cm reaction coil to the flow cell and the fluorescence intensity was measured. This signal is related only to cysteine. Thus, the ascorbic acid content was determined directly by the difference according to the calibration curve. Ascorbic acid and cysteine can be determined in the range of 1 x 10(-6) to 5.0 x 10(-5) M, at a rate of 16 samples per hour. The limits of detection (S/N = 3) were 8 x 10(-7) M for ascorbic acid and 7 x 10(-7) M for cysteine. The influence of potential interfering substances was studied. The proposed method was successfully applied to the simultaneous determination of both analytes in real samples. PMID- 16363476 TI - Toluene/ter-butanol mixed solvent for the selective extraction of Cr(VI) from divalent heavy metals. AB - A solvent-extraction system comprising toluene/ter-butanol (ter-BuOH) mixed solvent as the organic phase was developed to selectively extract Cr(VI) from acidic chloride media in the presence of divalent metals, namely Cd(II), Co(II), Cu(II), Ni(II) and Zn(II) under 5 M CaCl2 salting-out conditions. Chromium(VI) was selectively extracted as a solvated ion-pair of [ter-BuOH2+ x CrO3Cl-] at ter BuOH mole fractions of between 0.1 and 0.6 (9.0-57.2% in volume). Divalent metals were extracted at ter-BuOH mole fraction over 0.6 with extraction percents of Co (< 20%), Cu (< 15%), Ni (< 10%) and Zn (< 20%). The concentrations of Ca2+, water and ter-BuOH in the organic phase and ter-BuOH in the aqueous phase were determined to find out the effects on the extraction of Cr(VI). The chemical species of Cr(VI) in acidic chloride media containing 5 M CaCl2 and 0.1 M HCl was confirmed to be the CrO3Cl- species. The effects of the acid, salt concentrations in the aqueous phase and the solvent composition of a mixed organic solvent on the extraction of Cr(VI) were evaluated. Based on the above studies, the extraction mechanism was elucidated and the optimum extraction conditions were determined. PMID- 16363477 TI - Resolution of biparametric mixtures using bead injection spectroscopic flow through renewable surface sensors. AB - A new, sensitive and simple bead injection spectroscopy-flow injection analysis (BIS-FIA) sensor with spectrophotometric detection, using a commercially available flow-cell, is described to the determination of biparametric mixtures. As an analytical model, the metallic mixture Cu(II) and Zn(II) has been chosen. The flow-cell (Hellma 138-OS) is filled by injecting in the flow system 300 microl of a homogeneous bead suspension of an anion exchanger gel (Sephadex QAE A 25) previously loaded with the chromogenic reagent 2-carboxyl-2-hydroxy-5 sulfoformazylbenzene (Zincon). A sequential reaction of Cu(II) and Zn(II) with Zincon to form two complexes is performed on the bead sensing support and the absorbance is monitored at 627 nm, after two successive injections from the mixture solution. The sample containing these metal ions is injected into the first carrier (deionized water, pH 5.9), and Cu(II) selectively reacts with Zincon on the beads, developing the analytical signal. Then, 600 microl of 2 M HCl is injected to decompose the complex, and the carrier solution is changed. At pH 11 (second carrier) both Cu(II) and Zn(II) react with the chromogenic reagent, the absorbance now corresponding to both analytes. The eluent is again injected to descompose both complexes. After three analyses the sensing bead surface is not regenerated. Then, beads are automatically discarded from the flow cell by reversal of the flow, and instantaneously transported out of the system. So the procedure exploits the combination of the concepts of flow-through renewable sensors with bead injection spectroscopy. Using a sample volume of 1000 microl, the calibration graph for Cu(II) is linear over the range 0.05 to 1 microg ml(-1) and for Zn(II) from 0.1 to 1.8 microg ml(-1) in the presence of each other. RSDs (%) lower than 5% are obtained for both analytes. The sensor is satisfactorily applied to individual determination or mixture resolution in waters, pharmaceuticals, soils and human hair samples. PMID- 16363478 TI - Mass spectroscopic approach to amino acids formation processes by UV irradiation to simple organic molecules in aqueous solution. AB - We have studied amino acid formation by UV (193 nm) irradiation to organic molecules (amines, alcohols and amides) in aqueous solution. Among several types of detected amino acids, small aliphatic amino acids (Gly and alpha-, beta-Ala and alpha-, beta-, gamma-ABA) were quantitatively identified. Among these small aliphatic amino acids, certain amino acids were formed in its free form, even before hydrolysis, contrary to the results of UV irradiation to a gas mixture of CO, NH3, and H2O, where amino acids were hardly detected before hydrolysis. The species distribution of identified amino acids showed a dependence on the starting organic molecules, and also on the presence of ammonia. The formation processes of the identified small aliphatic amino acids were investigated with the aid of electrospray ionization (ESI) MS and MS/MS measurements of photoproducts. Possible formation processes of these amino acid precursors from each starting molecules are proposed. By identifying the amino acid precursor, which has a chiral carbon atom, a new possibility is suggested for asymmetric photosynthesis of amino acid from achiral organic molecules. PMID- 16363479 TI - Nano-SIMS analysis of Mg, Sr, Ba and U in natural calcium carbonate. AB - Concentrations of minor (Mg and Sr) and trace (Ba and U) elements in four natural calcium carbonate samples were first analyzed by inductively coupled plasma mass spectrometry (ICP-MS) after chemical dissolution and calibrated against a standard dolomite. Their homogeneities were checked by in situ laser ablation (LA) ICP-MS with 10-20 spots. The carbonate samples were measured by using a high lateral resolution secondary ion mass spectrometer (Nano-SIMS NS50). A approximately 4 nA O- primary beam was used to sputter a 5-6-microm diameter crater on the sample surface, and secondary positive ions were extracted for mass analysis using an accelerating voltage of 8 kV and a Mattauch-Herzog geometry. A multi-collector system was adjusted to detect 26Mg+, 43Ca+, 88Sr+, 138Ba+, 238U16O+ and 238U16O2+ ions at the same time. A resolving power of 2500-5000 at 10% peak height was attained by an entrance slit set at 40 microm, and each exit slit at 50 microm with adequate flat-topped peaks. The observed 26Mg/43Ca, 88Sr/43Ca, 138Ba/43Ca and 238U16O2/43Ca ratios agreed well with those measured by LA-ICP-MS. Foraminifera shells were analyzed at 5-6 microm scale by Nano-SIMS. There was a large variation of the Mg/Ca ratios, up to +/- 38%, even in a single fragment of the shell, suggesting that although the ratios provide a useful paleoceanographic proxy at bulk scale, they may reflect a more complex pattern at < 10 microm scale. PMID- 16363480 TI - Tris(2-methyl-8-quinolinolato)gallium(III) as a fluorescent probe for sensitive silanol-testing. AB - The peak shape of tris(2-methyl-8-quinolinolato)gallium(III) by reversed-phase high-performance liquid chromatography was found to be very sensitive to trace amounts of silanol groups on the surface of octadecylsylanized silica gel (ODS silica gel). The variation of the peak of the gallium(III) complex can be used as a probe of the residual silanol groups in an ODS column. The chromatographic peak parameters of the complex were compared with the silanol activities output by some silanol-detecting tests using nitrogen-containing compounds as probes. The comparison was performed with several commercially available ODS columns and laboratory-packed columns in which the amount of silanol groups was controlled by mixing fully endcapped ODS materials and a non-endcapped ODS material. The peak height was the most effective parameter among the peak parameters, and much more sensitive than the silanol-detecting tests using nitrogen-containing compounds, in detecting a trace amount of silanol groups that could not be detected by other silanol-detecting tests. PMID- 16363481 TI - Determination of cadmium, zinc, nickel and cobalt in tobacco by reversed-phase high-performance liquid chromatography with 2-(8-quinolylazo)-4,5 diphenylimidazole as a chelating reagent. AB - A sensitive and selective method has been developed for the simultaneous determination of cadmium, zinc, nickel and cobalt. The method is based on the chelation of metal ions with 2-(8-quinolylazo)-4,5-diphenylimidazole (QAI) and the subsequent reversed-phase (RP) high-performance liquid chromatographic separation and spectrophotometric detection of the metal chelates. The chelates were separated on an RP column with acetonitrile-water containing ethylenediamine tetraacetic acid and sodium acetate (pH 7.5). Though Zn(II) and Cd(II) chelates with azo compounds were generally labile in the RP column, these chelates with QAI were successfully detected. When analyses were carried out at 575 nm and at 0.001 absorbance unit full scale, the peak height calibration curves were linear up to 2.0 ng for Cd(II), 2.4 ng for Zn(II), 0.14 ng for Ni(II) and 0.72 ng for Co(II) in 100-microL injections, respectively; the detection limits (3sigma, three times of the standard deviation for the blank signal) for Cd(II), Zn(II), Ni(II) and Co(II) were 4.8, 24, 2.4 and 7.2 pg in 100 microL of injected solution, respectively. The proposed method was successfully applied to the analysis of tobacco without any preliminary concentration or separation. PMID- 16363482 TI - Rapid analysis of polychlorinated biphenyls in the gas phase with resonance enhanced two-photon ionization: optimal injection of ions into the ion-trap storage/time-of-flight mass spectrometer. AB - LI-IT-TOFMS (laser ionization/ion-trap storage/time-of-flight mass spectrometry) is expected to be a powerful tool for environmental monitoring. In the research reported here, real-time LI-IT-TOFMS measurements were carried out on gaseous 2-4 chlorinated PCBs in order to evaluate the applicability of an environmental monitoring method. With respect to ion-trap storage for PCBs, we found that the effect was due to the driving RF voltage on the ring electrode in the ion trap. For PCBs ions produced by laser irradiation, we observed that it was more efficient to reach the center of the ion trap by using a gated RF voltage rather than by using a continuous RF voltage. The ion trajectories in the ion trap were simulated by SIMION 7.0. We found that the voltage of the exit end cap electrode affected both the number of ions trapped and the orbit of ions inside the trap cell. Optimization of this parameter was performed using both simulated and experimental results. The achievable PCBs sensitivity for real-time (1 min) measurement using the LI-IT-TOFMS method was found to be in the pptV range (<0.01 mg/m3N) by means of a comparison with the conventional gas sampling/GS-MS method. A satisfactory proportional relationship was confirmed between the laser-based and conventional results. PMID- 16363483 TI - Fluorescence analysis of p-hydroxymethamphetamine in urine by thin-layer chromatography. AB - A highly-sensitive analytical method for the detection of p hydroxymethamphetamine (pOHMA) in urine is presented. The proposed method combines liquid-liquid extraction with acetonitrile and solid-phase extraction by thin-layer chromatography (TLC) with oxidation using potassium hexacyanoferrate(III) and sodium hydroxide to detect the fluorophor of pOHMA. The detection limit for pOHMA is 10 ng (n = 3). The analysis of pOHMA in forensic samples is successfully performed, without interference from endogenous fluorophors, yielding concentrations in the appropriate range for methamphetamine abusers. PMID- 16363484 TI - HPLC determination of acetaminophen in saliva based on precolumn fluorescence derivatization with 12-(3,5-dichloro-2,4,6-triazinyl)benzo[d]benzo[1',2' 6,5]isoindolo[1,2-b][1,3]thiazolidine. AB - A high-performance liquid-chromatographic method for the determination of acetaminophen in saliva has been developed. This method is based on the precolumn derivatization of acetaminophen with 12-(3,5-dichloro-2,4,6 triazinyl)benzo[d]benzo[1',2'-6,5]isoindolo[1,2-b][1,3]thiazolidine, a new fluorescence derivatization reagent for phenolic compounds. The resulting derivative of acetaminophen is separated by isocratic elution on a reversed-phase column, and is fluorometrically detected at an emission wavelength of 560 nm with an excitation wavelength of 540 nm. The detection limit (signal-to-noise ratio = 3) was 0.1 microg/mL in saliva. The proposed method permits a highly sensitive and simple determination of acetaminophen in a small amount of saliva without any sample purification. PMID- 16363485 TI - Bisphenol A analog-imprinted polymers prepared by an immobilized template on a modified silica microsphere matrix. AB - Bisphenol A (BPA)-recognizing imprinted polymers were synthesized using a template immobilized on silica where the template was grafted to aminopropyl silica. The silica-template conjugate was co-polymerized with a cross-linker (ethylene glycol dimethacrylate) and a functional monomer (4-vinylpyridine or methacrylic acid). The synthesized silica-polymer composites were treated with an aqueous NH4HF2 solution to dissolve the silica matrix. The 4-vinylpyridine-based imprinted polymer showed strong binding affinity to BPA and structurally related compounds having two hydroxyl groups at the 4,4'-position in the bisphenol structure. PMID- 16363486 TI - Method performance study of the determination of total nitrogen in soy sauce by the Kjeldahl method. AB - The objective of this collaborative study was to evaluate the proposed method for determining the total nitrogen in soy sauce by the Kjeldahl method submitted to the Codex Alimentarius Commission for endorsement in accordance with the protocol for the design, conduct, and interpretation of method-performance studies. The digestive conditions of the proposed method are the addition of 10 mL of H2SO4, 10 g (8 g by using a block digester) of K2SO4, and 1 mL of 20% CuSO4 x 5H2O and 80 min boiling period after the liquid is cleared by a heating device. Seventeen laboratories participated, analyzing five soy sauce samples as blind duplicates. Since the volume sampling method used in the JAS (Japanese Agricultural Standard) method showed lower accuracy of data because of the density of soy sauce, the method of sampling by weight was adopted as the proposed method. The total amount of outlier data was within acceptable limits for method-performance studies (< or = 22.2%). Lysine and ammonium sulfate recoveries for all laboratories were > or = 98% and > or = 99% respectively. The RSDr (repeatability relative standard deviation) values ranged from 0.4 to 1.3%, and the RSDR (reproducibility relative standard deviation) values were from 0.8 to 1.9%. HORRAT (RSDR/predicted RSDR) for the reproducibility showed 0.2 to 0.4, indicating acceptable precision of the method and excellent analytical performance. PMID- 16363487 TI - Spectrophotometric determination of tranexamic acid in pharmaceutical bulk and dosage forms. AB - In this study, a simple, fast, accurate and sensitive spectrophotometric method has been developed for the determination of tranexamic acid in bulk and pharmaceutical preparations. The method is based on the reaction of ninhydrin with the primary amino group of tranexamic acid in the basic medium at pH 8.0. The reaction produces a bluish-purple color which absorbs maximally at 565 nm. Beer's law was obeyed in the range of 3-40 microg ml(-1) with molar absorptivity of 5.093 x 10(3) L mol(-1) cm(-1). The effects of various factors such as temperature, heating time, concentration of reagent, color stability and interferences were investigated to optimize the procedure. The results have been validated analytically and statistically. The proposed method has been applied for the determination of tranexamic acid in bulk and pharmaceutical preparations with good results. PMID- 16363488 TI - A simple and rapid in situ preconcentration method for trace ammonia nitrogen in environmental water samples using a solid-phase extraction followed by spectrophotometric determination. AB - A simple and rapid in situ preconcentration method for the spectrophotometric determination of trace ammonia nitrogen in environmental water samples has been developed based on solid-phase extraction using a small column packed with octadecyl group-bonded silica gel (Sep-Pak C18 cartridge). A water sample was taken into a graduated syringe for easy and simple operation and prevention of contamination immediately after sample collection. Ammonia in the sample was reacted with hypochlorite and thymol to be converted into indothymol blue; then the formed indothymol blue was collected as an ion pair between indothymol blue and tetrabutylammonium ion on a Sep-Pak C18 cartridge. The indothymol blue on the cartridge was stable for 4 days. The retained indothymol blue was easily eluted with a mixture of methanol and 0.01 mol/l sodium hydroxide solution. The color intensity due to the indothymol blue was spectrophotometrically measured at 725 nm. The proposed method was successfully applied to environmental water samples such as river water. PMID- 16363489 TI - To buy or not to buy: image devices challenge budgets. PMID- 16363490 TI - Resource markets nudge IV prices up, survey says. PMID- 16363491 TI - Distributor worker carries out daring storm rescue. PMID- 16363492 TI - Supply chainers are doers--military or civilian. PMID- 16363493 TI - Who are the most popular suppliers? Neoforma awards top performers. PMID- 16363494 TI - RFID technology helps facilities keep track of people and assets. PMID- 16363495 TI - [Peptic ulcer: current status and perspectives]. PMID- 16363496 TI - [Guidelines in the management of H. pylori infection in Japan]. PMID- 16363497 TI - [Significance of the Japanese guideline for the management of gastric ulcer]. PMID- 16363498 TI - [Present state and future view of the indications for H. pyloni eradication therapy]. PMID- 16363499 TI - [Establishment of 2nd line treatment regimen for H. pylori infection--future prospect for the application for national health insurance system in Japan]. PMID- 16363500 TI - [New classification of gastritis: an important aspect of H. pylori]. PMID- 16363501 TI - [H. pylori infection and gastric cancer: Japanese intervention trial]. PMID- 16363502 TI - [Translocation of cagA protein via type IV secretion system]. PMID- 16363503 TI - [Relationship between Helicobacter pylori CagA-SHP-2 interaction and gastric cancer]. PMID- 16363504 TI - [Structure and intoxication of VacA]. PMID- 16363505 TI - [Induction of IL-8 production by PicA, PicB]. PMID- 16363506 TI - [Characteristics and biological activity of NapA(neutrophil-activating protein A)]. PMID- 16363507 TI - [TLR4/MD -2-dependent recognition of H. pylori LPS during gastric inflammation]. PMID- 16363508 TI - [Mechanisms of inflammation induced by H. pylori-HSP60]. PMID- 16363509 TI - [Mechanisms of action of tipa (TNF-alpha inducing protein), a new carcinogenic factor, released from H. pylori]. PMID- 16363510 TI - [Helicobacter pylori SabA adhesin in Japan]. PMID- 16363511 TI - [Effects of the interleukin-8 -251A/T polymorphism on the transcriptional activity and gastric atrophy in Helicobacter pylori infection]. PMID- 16363512 TI - [Relationship of interleukin (IL)-17 and IL-8 levels in gastric mucosal damage of the Helicobacter pylori infected gastric ulcer patients]. PMID- 16363513 TI - [REG (regenerating gene) protein--roles of REG protein in H. pylori-induced gastritis and gastric cancers]. PMID- 16363514 TI - [Survival strategy of Helicobacter pylori in gastric mucosal environment: dynamic aspect of reactive oxygen and nitric oxide metabolism]. PMID- 16363515 TI - [Role of nitric oxide derived from inducible nitric oxide synthase in H. pylori infected gastric mucosal injury]. PMID- 16363516 TI - [Protective role of thioredoxin-1 in Helicobacter felis-induced gastritis]. PMID- 16363517 TI - [Roles of cyclooxygenase-2 in gastric mucosal defense in Helicobacter pylori infected gastric mucosa]. PMID- 16363518 TI - [Plasma and gastric ghrelin levels in subjects with Helicobacter pylori infection]. PMID- 16363519 TI - [NF-kappaB and MAPK-signaling pathways contribute to the gene expression and host response induced by Helicobacter pylori infection]. PMID- 16363520 TI - [Helicobacter pylori induces matrix metalloproteinase expression]. PMID- 16363521 TI - [Relationship between IL-7 and IL-1beta concentrations and gammadelta T cells counts in H. pylori infected-gastritis]. PMID- 16363522 TI - [Immune and inflammatory reaction to Helicobacter pylori in the stomach]. PMID- 16363523 TI - [H. pylori infection and epithelial cell apoptosis]. PMID- 16363524 TI - [Sonic hedgehog expression in H. pylori-colonized gastric mucosa of Mongolian gerbils]. PMID- 16363525 TI - [Helicobacter pylori infection and endoscopic gastric findings among a young Japanese population]. PMID- 16363526 TI - [Transmission route of H. pylori]. PMID- 16363527 TI - [Epidemiology of intrafamilial H. pylori infection]. PMID- 16363528 TI - [Infection route of Helicobacter pylori and preventive strategy against its infection]. PMID- 16363529 TI - [Long-term follow-up of H. pylori infection--from gastritis to gastric cancer]. PMID- 16363530 TI - [New findings on Helicobacter pylori genes: an overview]. PMID- 16363531 TI - [Drug resistant H. pylori in Japan: general remarks]. PMID- 16363532 TI - [Yearly change in occurrence of antibiotic-resistant Helicobacter pylori isolates in Shinshu University Hospital during 1998-2002]. PMID- 16363533 TI - [H. pylori motility analysis]. PMID- 16363534 TI - [Diagnostic methods for H. pylori infection and health insurance]. PMID- 16363535 TI - [Problem of 13C-urea breath test in the diagnosis of H. pylori after eradication]. PMID- 16363536 TI - [Helicobacter pylori stool antigen test]. PMID- 16363537 TI - [Evaluation of newly developed office-based stool tests for detecting Helicobacter pylori]. PMID- 16363538 TI - [Diagnosis of H. pylori infection by using magnifying endoscopy]. PMID- 16363539 TI - [Drug sensitivity test for Helicobacter pylori]. PMID- 16363540 TI - [H. pylori infection and abdominal symptoms]. PMID- 16363541 TI - [Gastric ulcer is one of the diseases for which Helicobacter pylori eradication is recommended]. PMID- 16363542 TI - [Duodenal ulcer is one of the diseases for which H. pylori eradication is recommended]. PMID- 16363543 TI - [Gastric MALT lymphoma is one of the diseases for which H. pylori eradication is recommended]. PMID- 16363544 TI - [Stomach after endoscopic mucosal resection of early gastric cancer for which H. pylori eradication is recommended]. PMID- 16363545 TI - [Atrophic gastritis is one of the diseases for which h. pylori eradication is recommended]. PMID- 16363546 TI - [Gastric hyperplastic polyp is one of the diseases for which H. pylori eradication is recommended]. PMID- 16363547 TI - [Non-ulcer dyspepsia (NUD) is one of the diseases for which H. pylori eradication is recommended]. PMID- 16363548 TI - [Gastroesophageal reflux disease (GERD). Helicobacter pylori eradication improves pre-existing reflux esophagitis in patients with duodenal ulcer disease]. PMID- 16363549 TI - [Idiopathic thrombocytopenic purpura (ITP) and H. pylori eradication]. PMID- 16363550 TI - [Extra-gastric lymphoma of MALT type and H. pylori eradication]. PMID- 16363551 TI - [Coronary artery disease and H. pylori eradication]. PMID- 16363552 TI - [Infection with Helicobacter pylori and chronic idiopathic urticaria]. PMID- 16363553 TI - [Chronic pruritic skin diseases and H. pylori eradication]. PMID- 16363554 TI - [Iron deficiency anemia in adolescents and H. pylori eradication]. PMID- 16363555 TI - [Cap polyposis and Helicobacter pylori]. PMID- 16363556 TI - [Recent topics on important drugs for H. pylori eradication: Omeprazole]. PMID- 16363557 TI - [Recent topics on important drugs for H. pylori eradication: Lansoprazole]. PMID- 16363558 TI - [Recent topics on important drugs for H. pylori eradication: Rabeprazole]. PMID- 16363559 TI - [Recent topics on important drugs for H. pylori eradication: Pantoprazole sodium]. PMID- 16363560 TI - [H. pylori eradication package set (Lansap)]. PMID- 16363561 TI - [Recent topics on important drugs for H. pylori eradication: Amoxicillin (AMPC)]. PMID- 16363562 TI - [Recent topics on important drugs for H. pylori eradication: Clarithromycin (CAM)]. PMID- 16363563 TI - [Recent topics on important drugs for H. pylori eradication: Metronidazole (MNZ)]. PMID- 16363564 TI - [Recent topics on important drugs for H. pylori eradication: Minocycline (MINO)]. PMID- 16363565 TI - [Effect of acid suppression in subjects with Helicobacter pylori infection]. PMID- 16363566 TI - [Concomitant dosing of a H2 receptor antagonist (H2RA) with a triple therapy increases the cure rate of Helicobacter pylori infection]. PMID- 16363567 TI - [Quality of ulcer healing and rebamipide]. PMID- 16363568 TI - [Evaluation of diagnostic method for Helicobacter pylori infection]. PMID- 16363569 TI - [Triple therapy for eradication of Helicobacter pylori in Japan]. PMID- 16363570 TI - [Adverse effects of Helicobacter pylori eradication and its prevention]. PMID- 16363571 TI - [Recent trends in first-line therapy for H. pylori infection in the world]. PMID- 16363572 TI - [Is antimicrobial susceptibility testing necessary before second-line treatment for Helicobacter pylori infection?]. PMID- 16363573 TI - [Susceptibility to antibiotics and drug metabolism in patients with H. pylori infection refractory to the initial treatment--therapeutic strategy based on susceptibility to CAM and CYP2C19 polymorphism]. PMID- 16363574 TI - [Second-line eradication therapy for H. pylori infection--PPI+AMPC+MNZ triple therapy]. PMID- 16363575 TI - [High dose dual PPI/AMPC therapy for the treatment of Helicobacter pylori infection after failure of usual standard triple PPI/AMPC/CAM therapy]. PMID- 16363576 TI - [Anti-Helicobacter pylori quadruple therapy]. PMID- 16363577 TI - [PPI plus bismuth-based quintuple therapy]. PMID- 16363578 TI - [Comparison between one-week and two-week regimens of new triple therapy in H. pylori eradication]. PMID- 16363579 TI - [Cetraxate hydrochloride (Neuer) induces the effectiveness of new triple therapy in H. pylori eradication]. PMID- 16363580 TI - [Investigation about 3rd line eradication of H. pylori]. PMID- 16363582 TI - [Eradication therapy for the improvement of quality of life in cases of Helicobacter pylori-positive functional dyspepsia]. PMID- 16363581 TI - [Helicobacter pylori eradication therapy for children]. PMID- 16363583 TI - [Eradication therapy for the remnant stomach infected with Helicobacter pylori ]. PMID- 16363584 TI - [Effects of Helicobacter pylori eradication on gastritis in patients with chronic liver disease]. PMID- 16363585 TI - [Eradication of Helicobacter pylori in patients with end-stage renal disease undergoing dialysis treatment]. PMID- 16363586 TI - [Long-term follow-up after H. pylori eradication]. PMID- 16363587 TI - [Development of reflux esophagitis after H. pylori eradication]. PMID- 16363588 TI - [Gastroduodenal erosions observed after Helicobacter pylori eradication]. PMID- 16363589 TI - [Changes in atrophy and intestinal metaplasia of the stomach after Helicobacter pylori eradication]. PMID- 16363590 TI - [Peptic ulcer recurrent in patients following successful eradication of Helicobacter pylori]. PMID- 16363591 TI - [Effect of maintenance therapy by H2RA on the recurrence of gastric and duodenal ulcer and on the occurrence of reflux esophagitis, gastric erosions, and duodenal erosions after H. pylori eradication]. PMID- 16363592 TI - [Long-term outcome in patients with gastric MALT lymphoma after H. pylori eradication]. PMID- 16363593 TI - [Characteristics of gastric cancer occurring after Helicobacter pylori eradication]. PMID- 16363594 TI - [Esophageal adenocarcinoma after Helicobacter pylori eradication]. PMID- 16363595 TI - [Obesity after the cure of H. pylori infection]. PMID- 16363596 TI - [Development of obesity and hyperlipidemia after eradication of Helicobacter pylori]. PMID- 16363597 TI - [Effect of probiotics on the prevention of diarrhea in Helicobacter pylori therapy]. PMID- 16363598 TI - [High risk group for gastric cancer related with H. pylori infection]. PMID- 16363599 TI - [Helicobacter pylori-induced enlarged fold gastritis is associated with an increased risk of gastric carcinoma]. PMID- 16363600 TI - [Nodular gastritis and gastric cancer]. PMID- 16363601 TI - [Gastric adenoma, hyperlastic polyp and fundic gland polyp]. PMID- 16363602 TI - [Relationship between NSAIDs and H. pylori in peptic ulcer diseases--is the eradication of H. pylori recommended in NSAIDs ulcer?]. PMID- 16363604 TI - [Influence of interleukin-1beta genetic polymorphism on H. pylori eradication therapy]. PMID- 16363603 TI - [Influence of cytochrome P450 2C19 genetic polymorphism on H. pylori eradication therapy]. PMID- 16363605 TI - [Effect of lactoferrin on H. pylori colonization]. PMID- 16363606 TI - [Effects of sulforaphane-rich broccoli sprouts on H. pylori-infected gastric mucosa]. PMID- 16363607 TI - [Immunotherapy using dendritic cells for Helicobacter pylori infection]. PMID- 16363608 TI - [Antibacterial activity of a Chinese herbal medicine, Gosyuyu (Wu-Chu-Yu), against Helicobacter pylori]. PMID- 16363609 TI - [Prospects of development of H. pylori vaccines]. PMID- 16363610 TI - [Relation of Helicobacter heilmannii to gastric mucosal damage as a model of zoonosis between men and pets]. PMID- 16363611 TI - [Influence of Helicobacter pylori infection on the serum lipids and pulse wave velocity]. PMID- 16363612 TI - [Pathogenesis of Helicobacter pylori-negative peptic ulcer]. PMID- 16363613 TI - [Helicobacter pylori negative atrophic gastritis]. PMID- 16363614 TI - [Low prevalence of Helicobacter pylori infection in patients with fundic gland polyps]. PMID- 16363615 TI - [H. pylori-negative gastric MALT lymphoma]. PMID- 16363616 TI - Hemodynamic evaluation of varicocele: the role of scrotal scintigraphy and Doppler ultrasonography in the prediction of postoperative seminal improvement. AB - AIM: The aim of this study was to evaluate the hemodynamics of varicocele using Doppler ultrasonography and scrotal scintigraphy, and to compare the value of these two methods in the prediction of seminal improvement after varicocelectomy. MATERIALS AND METHODS: A total of 40 men with left sided varicocele presented for surgery because of infertility of at least one year in duration. Preoperative and postoperative sperm counts and per cent motility were obtained. Dynamic scrotal scintigraphy and Doppler ultrasonography were performed in all patients. Three perfusion patterns according to the time-activity curves (TAC) generated from scrotal perfusion images were defined. Type 1: radioactivity shows faster accumulation and maintenance of a higher level on the left side than on the right side. Type 2: time-activity curve rises gradually to a higher level on the left than on the right. Type 3: time-activity curve increases symmetrically and slowly on both sides. The relationship between preoperative TAC patterns and postoperative seminal findings, and preoperative Doppler grades and postoperative seminal findings were investigated. RESULTS: Improvement in total motile sperm counts was not statistically significant (37.8% +/- 3.2% versus 45.2% +/- 8.5%) (p = 0.751). Following varicocelectomy, sperm concentration (million sperm per ml) increased from 16.9 +/- 3.3 to 26.6 +/- 8.6 (p = 0.015). According to the Doppler examinations, postoperative improvement in sperm concentration was demonstrated in patients with grade 1 varicocele (66%). Scintigraphic evaluation showed improvement in patients showing TAC-2 and TAC-3 patterns (63%). CONCLUSION: Local hemodynamics of varicoceles demonstrated by scintigraphy and Doppler seemed to be different. Grade 1, TAC-2 and TAC-3 patients may be better candidates for varicocelectomy. Scintigraphy and Doppler ultrasonography showed similar success rates in the prediction of improvement following varicocelectomy in the present study. PMID- 16363617 TI - Ongoing myocardial damage relates to cardiac sympathetic nervous disintegrity in patients with heart failure. AB - Iodine-123-metaiodobenzylguanidine (123I-MIBG) has been used to assess the integrity and function of the cardiac sympathetic nervous system in patients with heart failure. Heart-type fatty acid binding protein (H-FABP) is released into the circulation when the myocardium is injured, and H-FABP has been recently used as a novel marker for the diagnosis of ongoing myocardial damage. OBJECTIVE: The aim of the present study was to compare cardiac sympathetic nervous activity assessed by 123I-MIBG imaging with serum levels of H-FABP in patients with heart failure. METHODS: Fifty patients with chronic heart failure were studied. 123I MIBG imaging was carried out at 30 min (early) and 240 min (delayed) after the tracer injection. We measured serum levels of H-FABP using a sandwich enzyme linked immunosorbent assay. RESULTS: Heart to mediastinum (H/M) ratios of 123I MIBG decreased and washout rate increased with higher New York Heart Association (NYHA) functional class. H-FABP, norepinephrine and brain natriuretic peptide (BNP) levels increased as the severity of NYHA class advanced. Delayed H/M ratio was significantly correlated with H-FABP (r = -0.296, p = 0.029) and BNP (r = 0.335, p = 0.0213). Myocardial washout rate of 123I-MIBG was also correlated with H-FABP (r = 0.469, p < 0.001), norepinephrine (r = 0.433, p = 0.005), and BNP (r = 0.465, p = 0.001). CONCLUSIONS: These data suggest that cardiac sympathetic nervous activation was associated with ongoing cardiomyocyte damage characterized by an elevated serum level of H-FABP in patients with heart failure. 123I-MIBG imaging is an appropriate approach to evaluate non-invasively not only cardiac sympathetic nervous activity, but also latent ongoing myocardial damage in the failing heart. PMID- 16363618 TI - Optimum energy window setting on Hg-201 x-rays photopeak for effective Tl-201 imaging. AB - For more effective Tl-201 imaging, the location and width of the energy window set on the Hg-201 x-rays photopeak was investigated using Monte Carlo simulation and phantom experiments. We calculated energy spectra and investigated the amount of primary and scattered photons within various energy windows set on the x-rays photopeak. The energy resolution (ER) at 71 keV (the peak of the x-rays photopeak) was changed to 10%, 12%, 14% and 16%. The relationships between the energy window and the primary counts rate or the scatter fraction (= scattered counts/primary counts, SF) were obtained. By compromise between the primary counts rate and the SF for ER = 12%, the optimum energy window was determined as a wider off-peak window, 77 keV +/- 14.3% (66-88 keV). This off-peak window increased the primary counts rate by 12.5% and decreased the SF by -17% as compared with the conventional on-peak energy window (71 keV +/- 10%, 64-78 keV). When this off-peak widow acquisition was compared with the conventional on-peak window one on a gamma camera, planar and SPECT images using the off-peak widow clearly showed superior results qualitatively and quantitatively. PMID- 16363619 TI - Evaluation of the hemodynamics of the femoral head compared with the ilium, femoral neck and femoral intertrochanteric region in healthy adults: measurement with positron emission tomography (PET). AB - OBJECTIVE: Non-traumatic osteonecrosis of the femoral head (ONF) is considered to be a disease that occurs primarily due to ischemia of the femoral head, while its etiology and pathology are not fully understood. It is therefore necessary to identify the characteristics of the hemodynamics of the femoral head. In this study, the hemodynamics in the ilium and proximal regions of the femur, including the femoral head, was investigated using positron emission tomography (PET). METHODS: The subjects of this study consisted of 8 hip joints of four healthy male adults and 3 hip joints on the contralateral side of a femoral neck fracture, avulsion fracture of the greater trochanter and coxarthrosis (1 case each, all females) for a total of 11 hip joints of 7 subjects. The ages of the subjects ranged from 25 to 87 years (average age: 54 years). Blood flow was measured by means of the H215O dynamic study method and blood volume was measured by means of the 15O-labeled carbon monoxide bolus inhalation method. RESULTS: Blood flow was determined to be 9.1 +/- 4.8 ml/min/100 g in the ilium and among proximal regions of the femur (femoral head, neck and intertrochanteric region), 1.8 +/- 0.7 ml/min/100 g in the femoral head, 2.1 +/- 0.6 ml/min/100 g in the femoral neck, and 2.6 +/- 0.7 ml/min/100 g in the intertrochanteric region. In addition, blood volume was 4.7 +/- 1.3 ml/100 g in the ilium, and among proximal regions of the femur, 1.1 +/- 0.5 ml/100 g in the femoral head, 2.1 +/- 0.7 ml/100 g in the femoral neck, and 2.6 +/- 0.9 ml/100 g in the intertrochanteric region. The results showed that both blood flow and volume were lowest in the femoral head. Blood flow and volume were significantly lower in the proximal regions of the femur (femoral head, neck and intertrochanteric region) than in the ilium (p < 0.01). CONCLUSION: The present study demonstrated that the femoral head is in a hypoemic state as compared with other osseous tissue, indicating that even the slightest exacerbation of hemodynamics in the femoral head can trigger an ischemic condition culminating in ONF. PMID- 16363620 TI - Do short-time SPECT images of bone scintigraphy improve the diagnostic value in the evaluation of solitary lesions in the thoracic spine in patients with extraskeletal malignancies? AB - OBJECTIVE: Single photon emission computed tomography (SPECT) images provide many details of the anatomical structure. Also about bone scintigraphy, there are many reports of the improvement of diagnosis by SPECT images. Although SPECT is useful, it requires much time. So to perform SPECT for all cases is difficult in the clinical situation. Recently, due to technical improvements in gamma cameras, we can get SPECT images in a short time. We examined diagnosis of solitary hot spots of thoracic spine in cancer patients using short-time SPECT. And we considered whether short-time SPECT contributes to the precise diagnosis of the lesion. MATERIAL AND METHODS: We performed bone scintigraphy image acquisition and both planar and short-time SPECT of the chest. Short-time SPECT was acquired in 6 minutes. We selected 36 cases with malignancy, whose bone scintigraphy demonstrated a solitary accumulation hot spot in the thoracic spine. Three experienced radiologists in nuclear medicine and 4 beginners diagnosed the images. They interpreted planar, short-time SPECT and maximum intensity projection (MIP) view of the chest of each case. The observers' response data were analyzed with receiver operating characteristic (ROC) curve analysis. RESULTS: Of the three types of images, the Az (the area under ROC curve) values of short-time SPECT were the highest in all the observers except for only one beginner. Compared with experienced observers, beginners scored lower Az values of short-time SPECT. MIP images were constructed using SPECT data, but the Az values of MIP images were not higher than those of planar images. As to diagnosis, beginners tended to interprete most of the accumulations as metastatic lesions. CONCLUSION: Short-time SPECT can be helpful to some degree, but to provide greater benefit, the observers require considerable exercise and experience. PMID- 16363621 TI - Influence of photon scattering and attenuation on ROI analysis in brain perfusion single-photon emission tomographic imaging of normal subjects. AB - OBJECTIVE: The aim of this study was to evaluate the effect of scatter and attenuation correction in region of interest (ROI) analysis in brain perfusion single-photon emission tomography (SPECT) and to assess the influence of selecting the reference area on semi-quantification. METHODS: Ten normal subjects were enrolled and injected with 123I-iodoamphetamine to undergo simultaneous emission and transmission scanning for scatter and attenuation correction. We reconstructed three SPECT images from common projection data of each subject: with scatter correction and non-uniform attenuation correction, with scatter correction and uniform attenuation correction, and with uniform attenuation correction applied to data without scatter correction. A program for automated ROI drawing was used to set ROIs on various regions in brain images. Regional count ratios were compared in images with different correction procedures by using three different reference areas. RESULTS: The effect of the combination of scatter and attenuation correction was marked in the precentral, temporal, posterior, hippocampus and especially in the cerebellum. In contrast, it was not appreciable in the central and parietal areas. When using the cerebellar ROI as the reference, the count ratio varied widely depending on the correction procedures. On the other hand, the whole brain reference offered the least variation in the count ratio. CONCLUSIONS: The influence of photon scattering and attenuation was dependent on regions. Since the count in the cerebellar ROI is greatly affected by photon scattering and attenuation, nonuniform attenuation correction combined with scatter correction deserves consideration when using the cerebellar ROI as the reference. PMID- 16363622 TI - Comparison of 18FDG-PET with 99mTc-HMDP scintigraphy for the detection of bone metastases in patients with breast cancer. AB - OBJECTIVE: Bone is one of the most common sites of metastasis in breast cancer patients. Although bone scintigraphy is widely used to detect metastatic breast cancer, the usefulness of 18FDG-PET for detecting bone metastasis has not been clearly evaluated. The purpose of this study was to compare the diagnostic accuracy of 18FDG-PET with bone scintigraphy in detecting bone metastasis in breast cancer patients. METHODS: Forty-four women aged 35 to 81 years (mean, 56 years) with breast cancer were examined in this study. Both 18FDG-PET and bone scintigraphy were performed for each patient with 0-69 day intervals (mean, 11.5 days). The results of each image interpretation were compared retrospectively. Whole-body bones were classified into 9 anatomical regions. Metastases were confirmed at 45/187 regions in 14 patients by bone biopsy or clinical follow-up including other imaging techniques for a period of at least 6 months afterwards. RESULTS: On a region basis, the sensitivity, specificity, and accuracy of 18FDG PET were 84%, 99% and 95%, respectively. Although these results were comparable to those of bone scintigraphy, the combination of 18FDG-PET and bone scintigraphy improved the sensitivity (98%) and accuracy (97%) of detection. False negative lesions of bone scintigraphy were mostly bone marrow metastases and those of 18FDG-PET were mostly osteoblastic metastases. 18FDG-PET was superior to bone scintigraphy in the detection of osteolytic lesions (92% vs. 73%), but inferior in the detection of osteoblastic lesions (74% vs. 95%). CONCLUSIONS: This study shows that 18FDG-PET tends to be superior to bone scintigraphy in the detection of osteolytic lesions, but inferior in the detection of osteoblastic lesions. 18FDG-PET should play a complementary role in detecting bone metastasis with bone scintigraphy. PMID- 16363623 TI - Tear clearance measurement in patients with dry eye syndrome using quantitative lacrimal scintigraphy. AB - OBJECTIVE: The aim of this study was to evaluate the tear clearance in patients with dry eye syndrome using quantitative lacrimal scintigraphy. MATERIALS AND METHODS: We investigated 21 patients (42 eyes; 18 women, 3 men; mean age, 63.19 +/- 13.33 years) with dry eye syndrome. Additionally, for the sake of comparison, 12 normal subjects of the same age group (24 eyes; 10 women, 2 men; mean age, 68.25 +/- 2.63 years) were included. Lacrimal scintigraphy, Schirmer-1 test, BUT, and rose bengal ocular surface vital staining were performed in these cases. RESULTS: According to the results of lacrimal scintigraphy, the mean value of T 1/2 was 4.16 +/- 1.22 minutes and the mean value of RI was 14.15% +/- 2.30% in normal subjects. However, in patients with dry eye syndrome, these values were 20.59 +/- 1.97 minutes and 55.64% +/- 6.90%, respectively. Consistent with the results of ophthalmologic tests, the mean Schirmer-1 value was 12.46 +/- 2.10 mm, the mean value of BUT was 14.36 +/- 3.40 seconds, and the mean staining value of the rose bengal was 1.98 +/- 0.80 in normal subjects, whereas these values were 1.36 +/- 0.49 mm, 5.46 +/- 1.33 seconds, 6.62 +/- 0.86, respectively, in patients with dry eye syndrome. When we compared the results of lacrimal scintigraphy and the results of ophthalmologic tests, an inverse correlation was noted between both the T1/2 and RI values and both the Schirmer-1 and BUT values in all subjects (p < 0.001). However, there was a greater positive correlation between the rose bengal ocular surface staining value and both the T1/2 and RI values in all cases (p < 0.001). CONCLUSION: In the current study, it was concluded that although the lacrimal drainage system was normal, tear clearance was significantly delayed in dry eye patients. With this study, we have shown that quantitative lacrimal scintigraphy, which is an objective, practical, and noninvasive method, appears to be useful for the assessment of the tear clearance in patients with dry eye syndrome. PMID- 16363624 TI - An experimental study on O-[18F]fluoromethyl-L-tyrosine for differentiation between tumor and inflammatory tissues. AB - OBJECTIVE: O-[18F]fluoromethyl-L-tyrosine (18F-FMT) is a recently developed tumor detecting agent with simple preparation and high radiochemical yields. The aim of this study was to assess the potency of 18F-FMT for differentiating tumor and inflammatory tissues using an animal model with an implanted tumor and experimentally induced inflammatory foci. METHODS: An ascites hepatoma cell line, AH109A, turpentine oil and Staphylococcus aureus were inoculated subcutaneously into Donryu rats as a tumor model, aseptic inflammation model and bacterial infection model, respectively. The biodistribution of radioactivity was assessed in rats at 5, 10, 30, 60, and 120 min after injection with 18F-FMT. Dual tracer whole-body and macro autoradiographies were performed 60 min after injection with a mixture of 18F-FMT and 2-deoxy-D-[1-14C]glucose (14C-DG). RESULTS: Tumor uptake of 18F-FMT was on average 1.27% injected dose per gram of tissue (%ID/g) and 1.43% ID/g at 30 min and 60 min, respectively and significantly higher than that in other normal tissues, except the pancreas (3.48% ID/g at 60 min). The uptakes in the aseptic and bacterial inflammatory tissues were very low and were not different from those of the background tissues. Dual tracer whole-body and macro autoradiographic studies showed that tumor uptake of 18F-FMT was clearly higher than uptake by the other tissues, while 18F-FMT accumulated much less both in aseptic and bacterial inflammatory tissues. In contrast, the 14C-DG images showed high accumulations not only in tumors but also in aseptic and bacterial inflammatory tissues. CONCLUSION: 18F-FMT seems to be a promissing tracer for the differentiation between tumor and inflammation because of higher specificity to tumor. PMID- 16363625 TI - Localization of chyle leakage site in postoperative chylothorax by oral administration of I-123 BMIPP. AB - The authors present a 71-year-old woman who had a right chylothorax after right upper lobectomy for lung cancer. As the chylothorax was considered to be due to thoracic duct injury at the time of operation, lymphoscintigraphy was performed by oral administration of I-123 beta-methyl-iodophenyl pentadecanoic acid (BMIPP). After visualization of the stomach and intestine, abnormal accumulation of the radiotracer was found initially around the right pulmonary hilum and then spread laterally in the upper pleural cavity, indicating chyle leakage in the region of the right pulmonary hilum. Scintigraphic finding was well correlated with the subsequent thoracoscopic observation, showing chyle leakage from a lymphatic tributary near its confluence to the thoracic duct at the level of the azygos continuation. The disruption site was ligated by video-assisted thoracoscopic-surgery procedure with successful termination of the chyle leakage. Lymphoscintigraphy by oral administration of I-123 BMIPP is thought to be a useful method for localization of chyle leakage in patients with chylothorax induced by thoracic surgery. PMID- 16363626 TI - Increased accumulations of N-isopropyl-p-[123I]-iodoamphetamine related to tumefactive multiple sclerosis. AB - We present a 21-year-old woman with tumefactive multiple sclerosis (MS) that exhibited a rapidly progressive course. There were multiple tumor-mimicking contrast-enhance lesions with surrounding edema and mass effect on magnetic resonance imaging. Both early and delayed brain single photon emission computed tomography (SPECT) with N-isopropyl-p-[123I]-iodoamphetamine demonstrated increased accumulations of the tracer and a high retention on the lesions. The SPECT findings represent a diagnostic pitfall for distinguishing MS from malignant brain tumors in patients with intracranial tumor-like lesions. PMID- 16363627 TI - Improvement in Tc-99m HMPAO brain SPECT findings during donepezil therapy in a patient with pure akinesia. AB - A 58-year-old man presented with a history of disturbance in initiating gait. His history revealed meningoencephalitis five years prior to admission. Neurological examination included gait disturbance as difficulty in initiation and a hesitating speech with many freezing episodes and micrographia Magnetic resonance imaging (MRI) showed diffuse hyperintensity of frontal subcortical white matter on T2 weighted images. He was diagnosed with PA. L-Dopa up to the dosages of 1000 mg/ day and selegiline 10 mg/day were given. First brain SPECT using technetium 99m labeled D,L-hexamethylpropylene amine oxime (Tc-99m HMPAO) was performed when he was taking L-dopa and selegiline. In visual evaluation, hypoperfusion in bilateral frontoparietal cortex was seen (Fig. 2). Treatment with L-dopa and selegiline produced no benefit. Donepezil 10 mg/day was begun. This therapy regimen resulted in dramatic clinical improvement within several days that was confirmed by blinded raters who watched the patient's video recordings. During this response second brain perfusion SPECT study was repeated during donepezil therapy. Markedly increased perfusion in bilateral frontoparietal cortex was observed. This is the first case of PA to develop possibly after an episode of bacterial pneumococcal meningoencephalitis and who responded to donepezil as documented by changes in clinical findings and Tc-99m HMPAO brain SPECT studies. PMID- 16363628 TI - 111In-pentetreotide and 123I-MIBG for detection and resection of lymph node metastases of a carcinoid not visualized by CT, MRI or FDG-PET. AB - A patient with a history of a jejunal carcinoid and resection of liver metastases underwent CT, MRI and FDG-PET as well as somatostatin receptor scintigraphy using 111In-pentetreotide during follow-up. Octreoscan demonstrated one extrahepatic abdominal lesion with pathologic uptake, while the other imaging modalities failed to show a corresponding abnormality. For verification of this finding 123I MIBG scintigraphy was performed. The MIBG scan confirmed the octreotide positive lesion and showed an additional abdominal lesion in the SPECT study. According to the scintigraphic results, radioguided surgery (RGS) was implemented using 123I MIBG. This resulted in the intra-operative detection of two para- and pre-aortic lymph node metastases by the gamma probe and successful resection. An additional preaortal lymph node, suspicious by palpation, was also removed. Histopathology revealed metastases of a carcinoid tumor in all three specimens. In conclusion, the use of RGS facilitates successful removal of carcinoid metastatic lesions despite negative conventional imaging results. Secondly, 123I-MIBG scintigraphy may provide advantages over octreoscan for preoperative localization as well as radio-guided surgery of neuroendocrine metastatic lesions, if the involved site is located in proximity to highly octreotide-avid organs such as the kidneys or spleen. PMID- 16363629 TI - Radiosynthesis and in vivo evaluation of 11C-labeled 1,5-diarylpyrazole derivatives for mapping cyclooxygenases. AB - We prepared 11C-labeled 5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3 (trifluoromethyl)-1H-pyrazole ([11C]1) and 4-[5-(4-methoxyphenyl)-3 trifluoromethyl-1H-pyrazol-1-yl]benzenesulfonamide ([11C]2) for imaging COX-1 and COX-2 isoforms, respectively, by positron emission tomography. [11C]1 and [11C]2 were synthesized in high radiochemical yields by O-[11C]methylation with [11C]methyl triflate in acetone containing an equivalent of NaOH as a base with respect to the phenolic precursors. In vivo evaluation in rats bearing AH109A hepatoma demonstrated minimal specific binding of [11C] to COX-1 in peripheral organs, such as the spleen and small intestine. Carrier-saturable uptake of [11C]2 was found in the spleen, but COX-2-specific binding of [11C]2 was not identifiable in the brain, AH109A hepatoma or other peripheral organs, although ex vivo autoradiography showed regionally different distribution in the brain and AH109A. The results suggest that neither [11C]1 nor [11C]2 is a suitable radioligand for in vivo biomarkers of COX enzymes, mainly because of marked non specific binding. PMID- 16363630 TI - Assessment of the solid-state gamma camera to depict axillary sentinel lymph nodes in breast cancer patients. AB - PURPOSE: The solid-state gamma camera is now commercially available offering the advantages of a compact and portable system, currently used mainly in the cardiac region. We evaluate the ability of the solid-state gamma camera to depict axillary sentinel lymph nodes (SLNs) in breast cancer patients. MATERIALS AND METHODS: Preoperative SLN lymphoscintigraphy (LSG) was performed in 19 patients with breast cancer using the solid-state gamma camera. Immediately thereafter, we performed a second LSG using a single detector Anger-type gamma camera, and compared the findings from the two cameras. RESULTS: Concordant results were obtained in 12 (63%) patients with both cameras. In 4 (21%) patients, axillary SLNs were correctly identified only with the solid-state gamma camera. In these patients, the distance between the SLN and the radiopharmaceutical injection site was closer than that of patients who had concordant results (p = 0.001). CONCLUSION: We can depict correctly axillary SLNs with the solid-state gamma camera in comparison with the Anger-type gamma camera. This technique would be useful for assessing SLNs in breast cancer patients. PMID- 16363631 TI - Evaluating performance of a pixel array semiconductor SPECT system for small animal imaging. AB - OBJECTIVES: Small animal imaging has recently been focused on basic nuclear medicine. We have designed and built a small animal SPECT imaging system using a semiconductor camera and a newly designed collimator. We assess the performance of this system for small object imaging. METHODS: We employed an MGC 1500 (Acrorad Co.) camera including a CdTe semiconductor. The pixel size was 1.4 mm/pixel. We designed and produced a parallel-hole collimator with 20-mm hole length. Our SPECT system consisted of a semiconductor camera with the subject holder set on an electric rotating stage controlled by a computer. We compared this system with a conventional small animal SPECT system comprising a SPECT 2000H scanner with four Anger type cameras and pinhole collimators. The count rate linearity for estimation of the scatter was evaluated for a pie-chart phantom containing different concentrations of 99mTc. We measured the FWHM of the 99mTc SPECT line source along with scatter. The system volume sensitivity was examined using a flood source phantom which was 35 mm long with a 32-mm inside diameter. Additionally, an in vivo myocardial perfusion SPECT study was performed with a rat. RESULTS: With regards to energy resolution, the semiconductor camera (5.6%) was superior to the conventional Anger type camera (9.8%). In the count rate linearity evaluation, the regression lines of the SPECT values were y = 0.019x + 0.031 (r2 = 0.999) for our system and y = 0.018x + 0.060 (r2 = 0.997) for the conventional system. Thus, the scatter count using the semiconductor camera was less than that using the conventional camera. FWHMs of our system and the conventional system were 2.9 +/- 0.1 and 2.0 +/- 0.1 mm, respectively. Moreover, the system volume sensitivity of our system [0.51 kcps/(MBq/ ml)/cm] was superior to that of the conventional system [0.44 kcps/(MBq/ml)/cm]. Our system provided clear images of the rat myocardium, sufficient for practical use in small animal imaging. CONCLUSIONS: Our SPECT system, utilizing a semiconductor camera, permits high quantitative analysis by virtue of its low scatter radiation and high sensitivity. Therefore, this system may contribute to molecular imaging of small animals and basic medical research. PMID- 16363632 TI - [Epidemiology of dystonias]. PMID- 16363633 TI - [Genetic dissection of dystonia]. PMID- 16363634 TI - [Pathology of dystonias: functional anatomy of the basal ganglia]. PMID- 16363635 TI - [Pathophysiology of dystonia]. PMID- 16363636 TI - [Treatment of dystonia]. PMID- 16363637 TI - [Reflex responses from the sural nerve to tibialis anterior muscle in hemiplegic patients: the relation between the responses and Babinski sign]. AB - Stimulation of the sural nerve of healthy subjects induced short latency inhibition in the ipsilateral tibialis anterior muscle and facilitation in peroneal muscle. We examined lower limb muscle responses after stimulation of the sural nerve in 19 patients with hemiplegia caused by cerebro-vascular disease and compared them with the control responses. The sural nerve was stimulated electrically (3 or 5 square wave pulses of 0.5 ms repeated at 250 Hz) during weak tonic contraction. Stimulation was triggered to average the rectified surface electromyography (EMG) of the test muscle. Usually 100 - 200 sweeps were averaged. After stimulation, the tibialis anterior muscle on the affected side of the hemiplegic patients showed the patterns of inhibition, facilitation, and no response, whereas all responses on the unaffected side, except those of one patient, were inhibition. The peroneal muscle on both sides showed only facilitation as in the controls. Abnormal responses of the tibialis anterior muscle on the affected side were present in many patients who had the Babinski sign. Abnormal responses in the tibialis anterior muscle of the affected side may have been due to contributions by disinhibition of the flexor reflex, late recruited motor units or both. PMID- 16363638 TI - [Temporal changes of diffusion-weighted MR images in a patient with transient global amnesia]. AB - The pathogenesis of transient global amnesia (TGA) remains uncertain, although it is featured by characteristic clinical symptoms. We present a patient with TGA, in which diffusion-weighted MR image showed a punctuate hyperintense lesion in the left CA1 subfield of the hippocampus adjoining the lateral ventricle 24 hours after the onset. This lesion was observed with FLAIR image after 9 days from the onset, and therefore was considered to be a delayed ischemic lesion. A SPECT and PET studies 7 days after the onset showed a decrease of the CBF, CMRO2 and CMRglu, and was compatible with a delayed ischemic lesion. However, there were no cardiovascular risk factors nor abnormalities of vascular supply, and therefore the reason for microcirculatory disturbances remained unclear. PMID- 16363639 TI - [A case of carbon monoxide poisoning by explosion of coal mine presenting as visual agnosia: re-evaluation after 40 years]. AB - We re-evaluated a case of carbon monoxide poisoning presenting as visual agnosia who had been injured by explosion of Miike-Mikawa coal mine 40 years ago. In an early stage, his main neuropsychological symptoms were visual agnosia, severe anterograde amnesia, alexia, agraphia, constructional apraxia, left hemispatial neglect and psychic paralysis of gaze, in addition to pyramidal and extra pyramidal signs. At the time of re-evaluation after 40 years, he still showed visual agnosia associated with agraphia and constructional apraxia. Concerning visual agnosia, recognition of the real object was preserved, while recognition of object photographs and picture was impaired. Thus, this case was considered to have picture agnosia as he could not recognize the object by pictorial cues on the second dimensional space. MRI examination revealed low signal intensity lesions and cortical atrophy in the bilateral parieto-occipital lobes on T1 weighted images. Therefore, the bilateral parieto-occipital lesions are likely to be responsible for his picture agnosia. PMID- 16363640 TI - [Familial amyotrophic lateral sclerosis with rapid progression]. AB - We report a family with autosomal dominant (AD) motor neuron disease. A 41-year old man developed muscle weakness and fasciculation of the lower extremities. The weakness progressed to the upper extremities and bulbar muscles. The cerebrospinal fluid (CSF) protein level was slightly elevated. A nerve conduction study revealed reduced compound muscle action potentials, but conduction block was not observed. Electromyogram showed acute and chronic neurogenic changes. He was treated with intravenous immunoglobulin (IVIg) and methylprednisolone pulse therapy, but his condition rapidly deteriorated. He developed respiratory failure necessitating artificial ventilation within three months after the onset of the disease. His father developed muscle weakness and atrophy of the upper extremities at age 70, and his cousin developed muscle weakness of the legs at age 41. Their conditions rapidly progressed to quadriplegia. CSF and electrophysiological findings were similar to those of the proband. Treatments by steroid pulse therapy, IVIg, and plasmapheresis were not effective. The father and cousin also required artificial ventilation within 3-4 months from the onset of symptoms, and became locked-in state. Autosomal dominant amyotrophic lateral sclerosis (AD-ALS) was considered, but SOD1 gene mutation was not detected. The present pedigree may have familial ALS caused by a gene mutation other than SOD1. PMID- 16363641 TI - [A 76-year-old woman with personality change, dementia and parkinsonism]. AB - We report a patient who developed personality change, dementia and parkinsonism. The patient was a Japanese woman who died at age 76. She developed memory problems at age 63. At age 66, she started showing personality changes, and began having short-step gait and mask-like face. On admission to our hospital at age 68, neurological examination showed mild memory deficit and postural instability. Six months after discharge, she developed delusion, rigidity, tremor, and gait disturbance. Her condition relentlessly progressed and she became bedridden at age 71. CT scan revealed marked atrophy of the frontotemporal lobes with enlargement of the lateral and third ventricles. The patient died at the age of 76 years. The patient was discussed in a neurological CPC, and a chief discussant arrived at the conclusion that the patient had frontotemporal dementia. Some participants thought that she had Pick disease or diffuse Lewy body disease. Severe atrophy of the frontal lobe and anterior part of the brain was seen at autopsy. Neuropathological examination showed severe neuronal loss with gliosis in the substantia nigra, pallidum, thalamus, and hippocampus. Moderate loss of neurons with gliosis was seen in the frontal and anterior temporal cortex. Argyrophilic and tau-positive neuronal inclusions which showed various shapes including Pick body-like inclusions and globose type of neurofibrillary tangles, were seen in the cerebral cortex and caudate. Argyrophilic and tau-positive astrocytes were also observed in the cerebral cortex. The pathological diagnosis was an unusual form of frontotemporal lobar degeneration with various tau positive inclusions. PMID- 16363642 TI - [Gene-protein dynamics generates circadian clocks]. PMID- 16363643 TI - [Stochastic fluctuations in cellular signal transduction systems]. PMID- 16363644 TI - [What is the origin of cell diversity?]. PMID- 16363645 TI - [Turing reaction-diffusion model in developmental biology: application to limb skeletal pattern formation]. PMID- 16363647 TI - [Translational control in cytoplasmic mRNP granules]. PMID- 16363646 TI - [Cancer initiation as mini-evolution]. PMID- 16363648 TI - [Molecular mechanisms of how we see]. PMID- 16363649 TI - [Mechanisms of body axis formation in the spider embryo: transformation from radial to bilateral symmetry]. PMID- 16363650 TI - [Cancer stem cells in cancer cell lines]. PMID- 16363651 TI - [The Japanese translation and cultural adaptation of Expanded Prostate Cancer Index Composite (EPIC)]. AB - PURPOSE: To develop a Japanese version of the Expanded Prostate Cancer Index Composite (EPIC): originally designed to measure the Quality of Life of localized prostate cancer patients, after careful assessments of cross-cultural equivalence, face validity and practically. METHODS: We translated the original version that consisted of 50 items into a preliminary Japanese version. This multi-stage procedure included a forward-translation, back-translation and discussion with the original developer. Additionally, we tested the preliminary Japanese version on 11 localized prostate cancer patients and identified problems with its cross-cultural equivalence, practicality. Based on the findings of this pretest, we revised the Japanese version. Consensus by discussion among all researchers was obtained through out this process. RESULTS: The original developer reviewed the back-translation of the preliminary Japanese version: some wording was revised. In the pretest, the average age of patients was 68.8 years old. Four of the sexual subscale showed over 10 percent missing data. In five items, all patients chose identical answers. We conducted an in-depth qualitative investigation of these items. The average response time was 11.7 minutes. We revised the Japanese to reflect patients' opinions as much as possible. Items which were showed problems in terms of cross-cultural adaptation included questions measuring 'bother' and two items of the sexual subscale. The wordings of these items were revised so that Japanese patients could easier understand them. We ensured that the original developer's intentions remained the same. The original developed approved all revisions. CONCLUSION: We translated and adapted the original EPIC to the Japanese culture. The Japanese version of EPIC was found to be functional in the pretest. PMID- 16363652 TI - [Pharmacological evaluation of efficacy and safety of propiverine hydrochloride in patients of overactive bladder--relationship between urodynamic observation and propiverine pharmacokinetics--]. AB - PURPOSE: The clinical benefit of propiverine hydrochloride against overactive bladder was evaluated, and the relationships between urinary voiding functions and the pharmacokinetics were investigated by means of clinical pharmacology with PK/PD approach. PATIENTS AND METHODS: Total 7 patients suffering urgency with urinary frequency and incontinence received propiverine hydrochloride in doses of 10 mg qd or 20 mg qd for 4 weeks, and then the doses were switched in cross-over manner to continue the treatment for further 4 weeks. The urodynamic measurements as well as pharmacokinetic samplings were done before the medication, 4 weeks and 8 weeks after the starting medication, to examine the dose-response and concentration-response relationships. RESULTS: The volume at first desire to void increased according to dose increased, and the volume at first involuntary contraction tended to increase according to both dose and drug concentration in plasma. However, no apparent dose-response relationships were observed for maximum urinary flow rate and the detrusor pressure at the maximum urinary flow rate. The PK/PD analysis using Emax model suggested that, approximately 75 ng/mL of the propiverine concentration in plasma allowed the increase in the volume at first involuntary contraction for 50%. The urinary residual volume increased in dose-dependent manner only in the patients with severe grade of lower urinary tract obstruction, but scarcely increased in the patients with moderate grade or below. CONCLUSION: Propiverine hydrochloride improved the urinary voiding functions with a tendency to depend on both dose and concentration in plasma. After the administration of propiverine hydrochloride, the concentration in plasma will immediately reach the level at which the drug can increase in the volume at first involuntary contraction for 50%, and then the concentration level will sustain the effect ranging from 10% to 50% increase in bladder volume. Furthermore, the lower urinary tract obstruction will be a predictor of increase in urinary residual volume. PMID- 16363653 TI - [Correlation of change in prostate-specific antigen and testosterone following withdrawal of androgen ablation after combination of radiation and hormone therapy]. AB - PURPOSE: We investigated retrospectively that change of serum testosterone and PSA concentrations following withdrawal of androgen ablation after combination of radiation and hormone therapy for prostate cancer. SUBJECTS AND METHOD: Among prostate cancer patients who were treated with combination of radiation and hormone therapy from 1992, 42 patients who were measured with time in the concentration of testosterone after withdrawal of androgen ablation were selected. Their median age was 76 years old (62-84), their median PSA was 13.2 ng/ml (1.4-215.3), and clinical stage consisted of T1 (12 patients), T2 (12 patients) and T3 (18 patients). They were divided into three groups by recovery of testosterone after withdrawal of androgen ablation and examined with change of PSA. Three groups consisted of castration-level group (testosterone: less than 1.0 ng/ml), low group (1.0-2.0 ng/ ml) and normal group (2.0 ng/ml and more). RESULTS: There were 8 patients in castration-level group, 10 patients in low group and 24 patients in normal group. There were no significant differences in age, clinical stage and histological grade among each group. However, duration of hormone therapy in normal group was 4.7 months and shorter than durations in castration-level (38.5 months) and low group (26.6 months). The median time of the recovery to normal range of testosterone was 9.1 months. Only duration of hormone therapy influenced the recovery of testosterone in the multivariate analysis. Median changes of PSA after withdrawal of androgen ablation were 0 ng/ml (0-0.029) in castration-level group, 0.118 ng/ml (0-1.169) in low group and 0.427 ng/ml (0.047-4.358) in normal group. Change of PSA in normal group was significantly higher than castration-level and low groups. If the patients were defined as failure in whom PSA was rising during follow-up, positive or negative predictive value to predict failure, when setting a cutoff value to 0.2 ng/ml of PSA rise-width, were better than other cutoff values (0.1 or 0.3 ng/ml). CONCLUSION: Among prostate cancer patients treated with combination of radiation and hormone therapy, PSA is went up in some of them with recovery of testosterone after withdrawal androgen ablation. Rise-width of PSA may be important for one of the predictors of failure with measurement of testosterone. PMID- 16363654 TI - [Prognostic factors for PSA relapse of prostate cancer after endocrine therapy]. AB - PURPOSE: Advanced prostate cancer responds well to endocrine therapy initially, but soon becomes refractory and has a poor prognosis. We analyzed the prognostic factors of prostate cancer responding well initially to endocrine therapy with lowering of serum prostate specific antigen (PSA) level but later showing PSA relapse. MATERIALS AND METHODS: In prostate cancer patients newly diagnosed from January 1992 to December 2004 at our institution, there were 93 patients in that the PSA level of 10 ng/ml or more before therapy initially dropped below 10 ng/ml by endocrine therapy, but showed PSA relapse thereafter. We investigated the relationship between clinical stage, pathological differentiation, initial PSA, duration between initiation of therapy and PSA nadir, the value of PSA nadir, duration between initiation of therapy and PSA relapse, PSA doubling time (PSA DT) at relapse, PSA response three months after initiation of second line therapy and prognosis after PSA relapse. RESULTS: In Kaplan-Meier method, between all or some categories investigated showed significant difference in prognosis after PSA relapse. In multivariate analysis, the factors that significantly affected prognosis after PSA relapse were clinical stage, pathological differentiation, PSA nadir value, duration between initiation of therapy and PSA relapse and PSA response three months after initiation of second line therapy. CONCLUSION: We investigated the prognostic factors refractory to endocrine therapy. These results are useful in planning the therapy, and in explaining the status or future prospective of the disease to patients and families. PMID- 16363655 TI - [Efficacy of periprostatic nerve blockade during ultrasound-guided transrectal 10 core biopsy of the prostate]. AB - PURPOSE: We examined the efficacy and safety of periprostatic nerve blockade during transrectal ultrasound guided prostate biopsy. MATERIALS AND METHODS: Transrectal ultrasound guided 10 core biopsy of the prostate was performed in 116 consecutive men. From March 2002 to July 2003, 58 men underwent biopsy of the prostate without local anesthesia (control group). From August 2003 to March 2004, 58 men received periprostatic nerve blockade before prostate biopsies (anesthesia group). A 4-ml dose of 1% lidocaine was injected at 2 or 3 locations on each side of the prostate via a 23 gauge needle. Pain during biopsy was questioned using a 5-point Face scale and complications were also recorded. RESULTS: The average pain score during biopsy was 1.9 in the anesthesia group versus 3.1 in the control group (p<0.001). In the anesthesia group 20.7% of patients had a pain score 3 or greater than 3 versus 69% in the control group. The complication rate showed no significant difference between the two groups. CONCLUSIONS: Periprostatic nerve blockade is a safe and effective method of anesthesia for transrectal prostate biopsy. PMID- 16363656 TI - [Prostate cancer metastasized to thyroid cartilage: a case report]. AB - A 75-year-old male was diagnosed as prostate cancer (serum PSA: 4,772 ng/ml, Gleason score: 4 + 4 = 8) with multiple bone metastases. And he noticed a painless mass of the frontal neck a month before the diagnosis. Computed tomography of the neck showed a tumor in the thyroid cartilage. Biopsy of the neck tumor revealed metastasis of prostate cancer by positive PSA staining. Metastasis of malignant tumor to cartilaginous tissue is extremely rare because there are usually no vessels in it. Only 4 cases of the metastasis of prostate cancer to the thyroid cartilage have been reported. It was thought that tiny bone marrows were formed in the ossified cartilage and it caused hematogenous metastasis. PMID- 16363657 TI - [A case of therapy-related leukemia/myelodysplastic syndrome following treatment of refractory testicular germ cell tumor]. AB - We report a patient with a refractory testicular non-seminomatous germ cell tumor (NSGCT) who developed therapy-related leukemia (TRL) after undergoing salvage chemotherapy and multiple operations for repeat recurrences. Fifty months after the initial therapy, pancytopenia and myeloblasts were observed in the patient's peripheral blood while the patient was undergoing salvage chemotherapy for a fifth recurrence. A bone marrow examination showed evidence of myelodysplastic syndrome (MDS) and refractory anemia with excess of blasts in transformation (RAEB in T) under French-America-British (FAB) classification. Cytogenetic 5q-/7q abnormalities were also observed. The patient had received a total dose of 189g/m2 of Ifosfamide, 8,250mg/m2 of Etoposide and 1,450 mg/m2 of Cisplatin; therefore, he was diagnosed as having TRL/MDS. The patient has received induction chemotherapy for TRL with Cytarabine, Daunorubicin and Fludarabine while a bone marrow transplantation has been scheduled. Recently, TRL associated with chemotherapy are being reported with increasing frequency in the literature. Since early detection and treatment are necessary for the management of TRL, peripheral blood examinations should be performed after a diagnosis of refractory germ cell tumor has been made. If pancytopenia is detected, bone marrow and cytogenetic examinations should be immediately performed to rule out TRL. PMID- 16363658 TI - [Localized amyloidosis of the bladder treated effectively by occlusive dressing technique therapy using Dimethyl sulfoxide (DMSO): two case reports]. AB - The first case is a 50-year-old female. She was consulted to our hospital for further examination of right hydronephrosis that was shown by ultrasonography. Cystoscopy revealed tumorous lesion around the right orifice. Bladder tumor was suspected and transurethral biopsy of the bladder was performed. The second case is a 54-year-old man. He was consulted to our hospital because right hydronephrosis was shown by counterized tomography. A tumorous or stenotic lesion in the lower end of ureter was suspected. Cystoscopy revealed yellowish and thickened tissue in the trigonal area. Bladder amyloidosis was suspected and transurethral biopsy of the bladder was performed. The histopathological diagnosis indicated amyloidosis and systemic amyloidosis was excluded. Both cases were treated by occlusive dressing technique therapy using Dimethyl sulfoxide (DMSO) successfully. PMID- 16363659 TI - [Solitary fibrous tumor presenting as perivesical mass associated with hypoglycemia: a case report]. AB - Some non-islet tumors can induce hypoglycemia. We report a case of a solitary fibrous tumor (SFT) as perivesical mass associated with hypoglycemia. A 61-year old man was admitted to our department with recurrent hypoglycemic attack and lower abdominal discomfort, but no symptom of urination. Ultrasonography and magnetic resonance imaging demonstrated a giant heterogeneous mass in the pelvic and rightward shift of bladder, but the tumor's border was clear. Endocrinological analyses showed high serum levels of insulin-like growth factor II (IGF-II) and suppressed secretion of insulin. We removed perivesical tumor that weighed 720 g on 30th January in 2003. After the removal, serum IGF-II levels returned to normal and hypoglycemic attacks ceased. Pathological examination revealed well-circumscribed nodular mass composed of uniform spindle cells arranged in bundles and fascicles with varying amounts of collagen and reticulin fibers. Almost all of the tumor cells were immunohystochemically positive for vimentin and CD34, negative for cytokeratin. PMID- 16363660 TI - [Metastatic tumor of spermatic cord with elevation of serum human chorionic gonadotropin beta-subunit]. AB - A 62-year-old man was presented with a firm mass in right scrotum. Serum LDH and AFP were within normal range, but hCG-beta was elevated (2.3 ng/ml). Under the diagnosis of right testicular tumor, he underwent right radical orchiectomy. The specimen was a spermatic cord tumor with poorly differentiated adenocarcinoma. hCG-beta was still elevated postoperatively and gastric fiber revealed adenocarcinoma of the stomach. Histochemical staining for hCG-beta was positive in both tumor of the spermatic cord and stomach. Finally the tumor was diagnosed as metastatic tumor of spermatic cord from gastric cancer, causing the elevation of hCG-beta. PMID- 16363661 TI - [Subjective assessment of quality of life and quality of vision]. PMID- 16363662 TI - [Expression and function of receptors for advanced glycation end products in bovine corneal endothelial cells]. AB - Corneal endothelial cell loss is a change that occurs with age, but its mechanism is still unclear. We postulated that interaction between advanced glycation end product(AGE) and its receptors is implicated in the corneal endothelial cell loss with age. We investigated the expression of AGE receptors: receptors for AGE(RAGE) and galectin-3 in bovine corneal endothelial cells by reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry. In addition, we investigated the effect of AGE on the cultured corneal endothelial cells. Expression of RAGE and galectin-3 was detected in bovine corneal endothelial cells. Galectin-3 was important in the internalization of AGE. In contrast, RAGE was important in the generation of reactive oxygen species and induction of apoptosis. Based on these data, the interaction of AGE in aqueous humor and AGE receptors expressed on the corneal endothelial cells was speculated to have a role in the corneal endothelial cell loss with age. PMID- 16363663 TI - [The immunoregulatory role of local antigen-presenting cells in ocular inflammation]. AB - The immune privilege that exists in the eye is maintained by various mechanisms. One of the best studied is a form of systemic tolerance termed anterior chamber associated immune deviation (ACAID). We have recently shoun that the vitreous cavity(VC) also had the ability to induce tolerance and named this phenomenon 'vitreous cavity-associated immune deviation(VCAID). Ocular antigen-presenting cells(APCs) are known to be critical in inducing both ACAID and VCAID. In contrast to normal conditions, an inflamed eye no longer supports either ACAID or VCAID induction. We therefore elucidated the mechanism for terminating already established ocular inflammation. Murine experimental autoimmune uveitis (EAU) is a model of human uveitis. Ocular-infiltrating macrophages produce various cytokines/chemokines and damage tissue in EAU. We found that only the macrophage enriched cells from the eye produced RANTES(regulated upon activation normal T cells expressed and secreted) Neutralization of RANTES by specific antibodies in vivo exacerbated EAU. We also found that the ratio of ocular CD 4/CD 8 T cells was markedly increased after treatment. As a result, RANTES neutralization might exacerbate EAU by modulating the type of T cell subsets recruited to the eye. Our data provide insight into the immunoregulatory role of macrophages mediated by RANTES in the pathogenesis of ocular inflammation. Not all macrophage-derived chemokines may cause local inflammation, since RANTES produced by ocular macrophages appears to suppress experimental autoimmune uveitis. PMID- 16363664 TI - [Mechanisms of retinal photoreceptor cell fate determination]. AB - PURPOSE: Understanding the molecular mechanisms by which distinct cell fate is determined during organogenesis is a central issue in cell development and disease. Using conditional gene ablation, we show that in mice the transcription factor Otx2 is essential for retinal photoreceptor cell fate determination and the development of the pineal gland. METHODS AND RESULTS: In the Otx2 knockout mice, differentiating photoreceptor cells were converted to amacrine-like neurons, and the pinealocytes in the pineal gland were totally absent. In addition, promoter assays revealed that Otx2 transactivates Crx, which is required for terminal differentiation and maintenance of photoreceptor cells. Furthermore, retroviral gene transfer of Otx2 steered retinal progenitor cells toward becoming photoreceptors. CONCLUSION: Thus, Otx2 is a key regulatory gene for the cell fate determination of retinal photoreceptor cells. Our results revealed the key molecular steps required for photoreceptor cell fate determination and pineal gland development. PMID- 16363665 TI - [Role of corneal fibroblasts in the pathogenesis of ocular allergic diseases]. AB - The accumulation of eosinophils and the subsequent release and deposition of cytotoxic proteins by these cells are thought to play an important role in the development of corneal lesions in individuals with vernal keratoconjunctivitis(VKC). Stimulation of corneal fibroblasts with the T helper 2 (Th2) type cytokines, interleukin (IL) -4 or IL-13, induces expression of both the chemokine eotaxin and vascular cell adhesion molecule-1, which together mediate eosinophil infiltration into the cornea. Corneal fibroblasts express a high-affinity receptor complex for IL-4 and IL-13, and their stimulation by these cytokines also induces expression of thymus- and activation-regulated chemokine(TARC), which is a potent chemoattractant for Th2 cells. Dermal fibroblasts, but not lung fibroblasts, also express TARC, suggesting that corneal, dermal, and lung fibroblasts play different roles in the initiation of corresponding Th2 cell-mediated allergic conditions. Corneal fibroblasts thus not only maintain tissue structure but also contribute to the induction and amplification of ocular allergic inflammation. These observations suggest that the development of drugs that specifically inhibit the functions of corneal fibroblasts may provide a basis for new treatments for the corneal disorders associated with VKC. PMID- 16363666 TI - [Quantitative determination of vitamin A levels in frozen preserved human amniotic membrane]. AB - PURPOSE: Retinoids are associated with cell proliferation and differentiation. The levels of retinoids (vitamin A and its derivatives) in human amniotic membrane (AM) were measured to investigate whether endogenous retinoids in AM might contribute to the process of wound healing of the keratoconjunctival epithelium in AM transplantation. SUBJECTS AND METHODS: Retinoids were extracted from AM obtained from three patients following cesarean section. Retinoids including retinol (ROL), retinylaldehyde (RAL), retinoic acid(RA), and retinyl palmitate(RP) were analyzed using a reverse-phase high performance liquid chromatograph equipped with a multi-wavelength detector or fluorescence detector. RESULTS: ROL was identified and detected in AM at a concentration of 9.42 2.75(mean +/- standard deviation) ng/g wet tissue. No other retinoids, were detected. CONCLUSION: Among the retinoids tested, only retinol was detected at low levels in human AM tissues. Based on these results, endogenous retinoids in AM probably have little effect on wound healing of the keratoconjunctival epithelium in AM transplantation. PMID- 16363667 TI - [The VF-14 index of functional visual impairment in patients with corneal disease treated by phototherapeutic keratectomy]. AB - PURPOSE: To evaluate the Visual Function 14(VF-14) index of functional visual impairment in patients with corneal disease treated by phototherapeutic keratectomy (PTK). METHODS: Seventy-two patients with corneal disease who were treated by PTK participated in this study between March 1999 and September 2002. Demographic, ocular history, best-corrected visual acuity, and detailed ocular examination data were collected. Functional visual impairment information was obtained by questionnaire covering the following: VF-14, Cataract Symptom Score, and satisfaction with vision. RESULTS: The mean best-corrected visual acuity of the eye scheduled for PTK was 0.67+/-0.63(mean +/- standard deviation) the logarithm of the minimum angle resolution (logMAR) whereas that of the post operated eye was 0.34+/-0.38 logMAR. The mean VF-14 score, Cataract Symptom Score, and satisfaction with vision were improved significantly after the treatment. The satisfaction with vision was correlated with the VF-14 score and the Cataract Symptom Score. The VF-14 scores of all of the patients whose pre-PTK VF-14 scores were under 50 were improved after PTK. CONCLUSIONS: The VF-14 is a valid measure of functional visual impairment in patients with corneal disease treated by PTK. PMID- 16363668 TI - [Anterior segment fluorescein angiography for evaluating the effect of vitrectomy for neovascular glaucoma]. AB - PURPOSE: Using anterior segment fluorescein angiography(AFA), we evaluated the relationship between the effect of vitrectomy combined with endophotocoagulation for neovascular glaucoma and the postoperative angiographic changes. SUBJECTS AND METHODS: Seven eyes of 7 patients (six eyes of 6 patients had proliferative diabetic retinopathy and 1 eye of 1 patient had central retinal vein occlusion) with neovascular glaucoma underwent both vitrectomy combined with endophotocoagulation and AFA before and after surgery. RESULTS: Of the 7 cases, the three eyes(42.9%), ther received one vitreous surgery alone had a reduction in intraocular pressure (IOP). The other 4 with uncontrollable IOP underwent trabeculectomy additionally, and three of them achieved good control of IOP. AFA showed less dye leakage in the iris after surgery in all cases. Three of 4 eyes with less dye leakage in the chamber angle showed good IOP control. Two of 3 eyes without the change in dye leakage were treated additionally by trabeculectomy. CONCLUSION: AFA is a sensitive technique in the assessment of neovascularization in the anterior segment, so that it is useful in evaluating the effect of vitrectomy combined with endophotocoagulation for neovascular glaucoma. PMID- 16363669 TI - [Retinopathy during interferon treatment in combination with ribavirin for chronic hepatitis C]. AB - PURPOSE: To observe the retinopathy of the patients who received interferon/ribavirin for treatment of chronic hepatitis C. METHODS: We observed 6 patients(5 males and 1 female) who received interferon/ribavirin for treatment of chronic hepatitis C. Visual acuity tests and detailed fundus examinations were performed monthly during 6 months of interferon/ribavirin therapy. RESULTS: All patients developed soft retinal exudate and 5 developed retinal blot hemorrhage. None of the patients exhibited visual impairment or subjective symptoms during the treatment period, and the retinopathy disappeared or decreased in all patients. All of the patients in this study developed interferon retinopathy while undergoing interferon/ribavirin combination therapy. CONCLUSION: Because the combination of ribavirin with interferon may increase the incidence of interferon retinopathy, and cases of severe retinal complications have also been reported, careful fundus examinations should be performed during combination therapy, just as they are performed during conventional interferon therapy. PMID- 16363670 TI - [Influence of cataract and intraocular lens surgery on health-related quality of life]. AB - PURPOSE: To evaluate the vision-related quality of life(QOL) in patients undergoing cataract surgery with the Japanese version of the 25-item National Eye Institute Visual Function Questionnaire (NEI VFQ-25). SUBJECTS AND METHODS: Subjects were 110 patients [70.4 +/- 9.2 (mean +/- standard deviation) years old] undergoing bilateral cataract surgery. Best-corrected visual acuity was 20/30 or worse in both eyes. The VFQ-25 was recorded before and 2 months after phacoemulsification and foldable intraocular lens implantation, and the influence of various clinical parameters was assessed. The VFQ-25 was also recorded in 69 glaucoma patients with visual field defects in both eyes, and in 31 normal subjects. RESULTS: The VFQ-25 scores before cataract surgery were as low as the scores obtained in glaucoma patients. Cataract surgery significantly improved VFQ 25 scores to the level of normal subjects Statistically significant improvements were observed in subscales such as general vision, near activities, distance activities, driving, peripheral vision, color vision, social functioning, dependency, role difficulties, mental health, and total score (p<0.001, paired t test). The degree of improvement in VFQ-25 scores did not correlate with preoperative or postoperative visual acuity, uncorrected or corrected. The VFQ-25 scores improved more in younger patients, and improvement in the general vision subscale showed a significant negative correlation with age (r= -0.286, p=0.009, Pearson correlation coefficient). The degree of posterior subcapsular cataract in better eyes showed significant correlation with the improvement in subscales, such as distance activity (r =0.413, p<0.001), driving (r= 0.449, p=0.015), social functioning (r=0.308, p= 0.004), mental health (r=0.330, p=0.002), dependency (r=0.323, p=0.003), and total score (r=0.328, p=0.002). Other types of cataract had no correlation. Men tended to show more improvement in VFQ-25 scores by surgery than women, except for the dependency subscale. CONCLUSION: The current study quantitatively demonstrated that vision-related QOL is significantly impaired in patients with cataract, and that cataract surgery dramatically improves patients' QOL. PMID- 16363671 TI - [A case report illustrating the effectiveness of reading performance assessments in the diagnosis and treatment of psychogenic visual disturbances]. AB - PURPOSE: An optimal strategy for treating psychogenic visual disturbances in adults has not been established. We report a patient with psychogenic visual disturbances who recovered his visual acuity and showed an improvement in his reading performance after undergoing training based on a reading performance assessment. CASE: A 37-year-old man who had been diagnosed as having myopic macular degeneration was referred to our clinic. Three months after his initial diagnosis, no changes in his fundi were observed, but his visual acuity had significantly decreased and his peripheral field of vision had become severely restricted. In view of his tunnel vision, the discrepancy among the visual acuity results obtained by different test methods, the results of a reading assessment, objective eye examination data, and his behavioral patterns, we diagnosed a psychogenic visual disturbance in the patient and referred him to an ophthalmologist and a psychiatrist for follow-up care. In our low vision clinic, we assessed his visual function, including reading performance, and developed a training program including reading, writing, and computer skills. We also provided information to help the patient find a job. The training program included instructions on how to manipulate reading aids and how to select reading materials to maximize his vision; these instructions were effective. Nine months after his rapid decrease in visual acuity, the results of his visual function tests showed an improvement. The patient also became motivated to find a job. CONCLUSION: Reading assessments are a useful tool for diagnosing psychogenic visual disturbances in adults and for coping with functional vision impairment. PMID- 16363672 TI - [Cost-effectiveness analysis of QuantiFERON-TB 2nd generation used for detection of tuberculosis infection in contact investigations]. AB - PURPOSE: QuantiFERON-TB-2nd Generation (QFT) has recently been developed as an accurate tool for detecting tuberculosis infection regardless of past history of BCG vaccination. A cost-effectiveness analysis was made on the usefulness of QFT that was used in the contacts investigation of a group of subject exposed to tuberculosis infection. METHODS: A model was built assuming that a group of youngsters was exposed to an infection source with different degrees of intensity. The distribution of the tuberculin reaction of this group was assumed to be variable according to the history of BCG vaccination and tuberculin testing. Also, the distribution of tuberculin reaction size after the recent exposure is assumed to be different, as has been observed previously. The strategies for investigating this group included giving QFT to subjects having erythema size exceeding 30 mm, 20 mm, and 10 mm as compared with the strategy with the tuberculin test only, or the QFT only. The outcome variables calculated for each strategy were sensitivity and specificity, and predictive values in detecting tuberculosis infection; the number of indications for chemoprophylaxis, the number of tuberculosis patients averted, and the costs incurred in treating tuberculosis patients and chemoprophylaxis cases and testing with tuberculin and QFT were also considered. The sensitivity (specificity) of the QFT employed in the analysis was 89% (98%) based on our observations. RESULTS & CONCLUSION: It was confirmed that the additional use of QFT would greatly reduce the number of indications for chemoprophylaxis cases that have never been infected and that the use of QFT is cost effective in spite of its relatively high unit cost. It will be useful to decide on the eligibility of QFT testing, i.e., the minimal tuberculin reaction size of subjects to whom QFT is given, based on the assumption of pre-exposure distribution of tuberculin reaction size of the group. PMID- 16363673 TI - [Treatment results of multi drug resistant tuberculosis, a hospital based study]. AB - SETTINGS: Fukujuji Hospital, Japan. PURPOSE: To evaluate treatment results of multi drug resistant tuberculosis cases. METHOD: Retrospective review of 100 multi drug resistant tuberculosis cases who started treatment in 1990-1999 at Fukujuji Hospital. Life table analysis and analysis of longterm treatment results were done. RESULT: Observation for more than 3 years after starting treatment revealed cure in 62, chronic excreters in 2, and died cases with positive culture in 10, and treatment completed with shorter observation period in 8. The remaining 18 cases were either transfer-out, defaulted or died with other causes. The older age, cavitary lesions (bilateral>unilateral>no), resistance to more drugs, interruption of treatment due to side effects to second line drugs and the complication of diabetes mellitus were independent risk factors for unfavorable results. The treatment results improved with the increase of the number of effective drugs used for 6 months or more. The surgical intervention improved the proportion of favorable treatment results for those that were culture positive after 5 months of treatment. DISCUSSION: Treatment results of MDR TB was not satisfactory. Diabetics was the risk factor of unfavorable results. The conditions for the indication of surgical intervention among culture negative cases after 5 months of medical treatment needs to be further investigated. PMID- 16363674 TI - [A case of tuberculous peritonitis]. AB - We report a case of a 73-year-old man with tuberculous peritonitis. He had sought treatment at a clinic near his house for his fever and abdominal distension. Massive ascites were found and he was referred to our hospital. The endoscopy and abdominal CT scan performed on admission revealed no abnormal findings except the massive ascites. Mycobacterium tuberculosis (MT) DNA was detected in the ascitic fluid by polymerase chain reaction (PCR) and ascitic adenosine deaminase (ADA) activity was 127.6 U/l. He was diagnosed as tuberculous peritonitis and transferred to the Department of Respiratory Medicine. A chest CT scan showed predominant right pleural effusion with no other abnormal findings in bilateral lung fields. His sputum were all positive by smear acid-fast staining, MT DNA and culture on MT. His final diagnosis was tuberculous peritonitis, pulmonary tuberculosis, and tuberculous pleuritis. Treatment was started by anti tuberculosis drugs with combined use of isoniazid, rifampicin, ethambutol, and pyrazinamide. The therapy was continued for 6 months. The culture for MT (Mycobacteria Growth Indicator Tube) converted to negative after 2 weeks of treatment and the C-reactive protein level became normal after a month. The pleural effusion and ascites disappeared after 2 and 3 months, respectively. Tuberculous peritonitis is a relatively rare disease, however when we encounter unexplained ascites, MT PCR and the measurement of ADA should be done considering a rapid diagnosis of tuberculous peritonitis, before invasive diagnostic laparoscopy. PMID- 16363675 TI - [Present situation and control on emerging respiratory infectious diseases such as SARS and avian influenza]. AB - Infectious diseases have been recognized again due to appearing of emerging and re-emerging infectious diseases in the world. Most of them occur not only in developing countries but also in developed countries, and in Asian region. The pathogen is mainly virus and most of them are suspected zoonotic origin. SARS emerged in the world abruptly and disappeared in 2003. We have had many lessons and learn on control measures, public health, economic impacts, human rights, international cooperation and infectious diseases. The outbreaks of avian influenza among fowls have been occurred since 2004, and some fatal human cases infected with avian influenza virus are detected in Viet Nam, Thailand, Cambodia and Indonesia. Although the total number of human cases are still limited and human to human transmission mode is not yet detected, it has been concerned the possibility to shift new types of influenza for human as pandemic. It is necessary to recognize correctly on existing of infectious diseases, to enhance surveillance, to call partnerships among several sectors such as medical institutes, medical education institutes, research institutes and public health departments. Further, infectious disease control should tackle in global level. PMID- 16363676 TI - [Viral zoonoses: introduction]. PMID- 16363677 TI - [The present status and points of viral zoonoses]. AB - In recent years, viral zoonoses are causing public health problems in Japan and abroad. This is due to the outbreaks of emerging viral zoonoses at different localities in the world. Due to the globalization, these emerging viral zoonoses may invade into Japan. To prevent the invasion and spread of the emerging viral zoonoses, quarantine measures should be strengthened against movements of man and animals. Diagnostic tests and vaccines to the zoonoses should be developed. Epidemiological survey must be conducted against man and domestic and wild animals in the endemic regions. International cooperation and information exchange are essential to establish the preventive systems for the viral zoonoses. PMID- 16363678 TI - [Present status, measure and future problems on the zoonoses from imported animals and vectors]. AB - Revision of the protection law for infectious diseases was conducted on 2004, which had been settled on 1999. Practical zoonosis control was one of the hot spot in this revision and policy making was done according to the scientific risk assessment. Thus, the newly settled measure is divided into more complex methods from simple ban or quarantine, to those proportionate to the risk levels. They are importation ban of the risk animals, mooring of wild birds, notification of certain zoonosis from veterinarian, notification with health certification papers for the imported animals, active surveillance and new vector control and so on. Present status, measure and future problems on zoonoses derived from imported animals and vectors are discussed. PMID- 16363679 TI - [Surveillance strategy for viral zoonotic diseases]. AB - Public health surveillance is the ongoing systematic collection, analysis, interpretation, and dissemination of health data. In fact, public health surveillance has been defined quite succinctly as "information for action". Recently, many outbreaks of virus zoonoses are reported in the world, which include West Nile virus disease, avian influenza and severe acute respiratory syndrome (SARS). Public health surveillance is a powerful tool for prevention. However establishing of public health surveillance systems are very difficult for random sampling, sample size and cost. The surveillance of Japanese encephalitis virus activity has been effectively conducted by using seroprevalence of pigs for more than 20 years in Japan. Polio acute flaccid paralysis (AFP) surveillance has been also effectively conducted, although polio cases are decreasing in the world. It is important to understand the character of disease for constructing effective public surveillance systems. PMID- 16363680 TI - [Human influenza--recent outbreak of avian influenza A/H5N1 among human in Asian countries--]. AB - Since late 2003, avian influenza A/H5N1 virus has infected 100 adults and children in Vietnam, Thailand, Cambodia resulting in 54 deaths (54.0%). The avian H5N1 is becoming more infectious for people and capable of human-to-human transmission. Experts have warned that the widespread occurrence of the H5N1 could ignite a pandemic capable of killing millions of people around the world. However, calculating with any precision the risk or when it will strike is impossible, and yet the consequences of failing to respond to this uncertain threat could be dire. At this stage, time-series epidemiological and virological analysis on the shift of avian to human influenza viruses are quite important for control of the H5N1 outbreak and the pandemic. PMID- 16363681 TI - [Avian influenza virus]. AB - Influenza viruses are classified as A, B, or C based on the antigenicity of their nucleoproteins and matrix proteins. Influenza A viruses are further categorized into subtypes based on the antigenicity of two of their surface proteins, the hemagglutinin and neuraminidase. All avian influenza viruses are type A viruses. While the majority of avian influenza viruses are avirulent, viruses of a limited number of subtypes cause severe disease in birds. This difference in pathogenicity among avian influenza viruses is primarily determined by the amino acid sequence at the viral hemagglutinin cleavage site. Since 1997, there have been multiple reports of human infection by highly pathogenic avian influenza viruses. Acquisition of the ability to cause human-to-human transmission by avian influenza viruses will confront the human population with an influenza pandemic crisis. Here we review the molecular basis of pathogenicity of avian influenza viruses in humans. PMID- 16363682 TI - [SARS coronavirus]. AB - SARS is a new type of infectious pneumonia that emerged in China in November 2002. It spread around the world through an outbreak in Hong Kong in March 2003. Patients were reported in 29 countries, and around 800 people died, although a majority of these cases were in Asian countries. Within a month after the virus that causes SARS was isolated, its entire genome sequence became available. The result is the acknowledgement of a novel coronavirus, now called SARS coronavirus (SCoV), that is different from other already known existing human and animal coronaviruses. Since a number of scientists from distinct fields have participated in the research on this emerging virus, a plenty of information has become available, and now SARS-CoV has become one of the best-studied members among coronaviruses. In this review, I would like describe on SCoV virology by comparison with other well-studied coronaviruses. It covers the taxonomy, structure of virion, viral genome and proteins, unique replication strategy, receptors and viral pathogenesis. In the last part, I mention with my personal speculation about the origin and evolution of SCoV. PMID- 16363683 TI - [Clinical and epidemiological review of SARS]. AB - Severe acute respiratory syndrome (SARS) was caused by a previously unrecognized new type coronavirus. The disease probably first emerged in Guangdong China around November 2002, and it spread from there first to countries in Southeast Asia, and finally throughout the world. During the outbreak, more than 8,000 cases and about 800 deaths had been reported. The first sign of illness in most cases of SARS is fever, headache, malaise, or myalgia, and then majority of patients present severe atypical pneumonia. It appeared that disease spread most often in hospitals in health care workers, and patients and among family members. Given the severity of SARS, clinicians throughout the world empirically treated most patients with corticosteroids and ribavirin. PMID- 16363684 TI - [West Nile fever]. AB - West Nile virus was first detected in North America in 1999 and has spread throughout the United States and Canada and into Mexico and the Caribbean. The cases of encephalitis in New York were diagnosed as Saint Louis encephalitis which was endemic in the North America. However, dead crows and dead flamingo were found around the same time in New York. American birds seldom die by Saint Louis encephalitis virus infection. According to viral isolation and sequencing of the genome, the causative agent of unknown encephalitis was West Nile virus which was a member of Flaviviridae which cause fever, meningitis and encephalitis. West Nile virus is still active in North America in summer season, 1999 -2005. CDC enlightens people how to protect themselves and their community from West Nile virus. There are some reports that West Nile viral infections were occurred by blood transfusion, biologic products from blood and organ transfusion. PMID- 16363685 TI - [Development of new Japanese encephalitis vaccine]. AB - We have developed the new inactivated Japanese encephalitis vaccine derived from virus-infected Vero cells cultured on microcarriers. The safety and effectiveness of this new vaccine were compared with those of current mouse-brain-derived vaccine that have some intrinsic demerits such as possible contamination of adventitious agents. The results of pre-clinical and clinical trials suggested that the safety and effectiveness of new vaccine are approximate equivalent to those of currently conventional vaccine in humans and animals, and that the availability of Vero cell-derived vaccine is anticipated. PMID- 16363686 TI - [Clinical and epidemiological aspects of Japanese encephalitis]. AB - Japanese encephalitis has been well controlled in Japan. On the other hand, the Japanese encephalitis virus (JEV) transmission cycle still exists, as does the risk of exposure to JEV infection. Although Culex tritaeniorhynchus has been considered the major mosquito vector in Japan, recent surveys suggested Culex pipiens complex and Aedes albopictus as potential alternatives. Similarly, swine have reduced the role in amplification of JEV in paradomestic environments. Another surveys suggested attenuation of recent JEV strains and a shift in the major clinical manifestation caused by JEV infection from encephalitis to meningitis. This paper describes current situation of Japanese encephalitis in Japan from clinical and epidemiological aspects. PMID- 16363687 TI - [Nipah virus infections]. AB - Nipah virus (NiV) is a zoonotic paramyxovirus that was first recognized in 1999 as the causative agent of outbreaks of human encephalitis in Malaysia and Singapore, in association with severe respiratory and neurological disease in pigs. Since then, outbreaks of NiV encephalitis have also occurred in Bangladesh during 2001-2004, but without an association to infected swine or other animals. Although NiV infections typically result in acute encephalitis with high mortality, other clinical manifestations, including asymptomatic infection, relapsed encephalitis, and pulmonary disease, have been observed. The article will summarize the virology, epidemiology, clinical features, treatment, and control and prevention of NiV infections in humans. PMID- 16363688 TI - [Countermeasure against viral hemorrhagic fever at the border in Japan]. AB - Human have struggled against many infectious diseases such as cholera, plague, dysentery and yellow fever for a long time. And we have spent a lot of energy to control these infectious diseases and developed various tool for them. One of these efforts was Quarantine system that was established in 14th century in Europe. But during recent days, we are suffering from newly emerged diseases. These new infectious diseases are zoonosis and most of them are serious and highly infectious. Viral hemorrhagic fever such as Ebola hemorrhagic fever, Marburg hemorrhagic fever and Lassa fever are typical these emerging serious diseases, and these outbreak always have occurred in Africa and neighboring countries. Fortunately we have never experienced any case, but as these diseases are so serious, we are so nervous diseases entering in Japan. Against these serious diseases, in Japan, Quarantine Station are doing screening examination at airport and port by questionnaire and measuring body temperature, because these viral hemorrhagic fever patients show high fever. If people were suspected viral hemorrhagic fever at Quarantine Station at the border, they will be leaded to hospital for further examination and treatment as soon as possible. PMID- 16363689 TI - [Clinical aspects of viral hemorrhagic fever]. AB - Viral hemorrhagic fever (VHF) is defined as virus infections that usually cause pyrexia and hemorrhagic symptoms with multiple organ failure. VHF includes following viral infections: Ebola hemorrhagic fever (EHF), Marburg hemorrhagic fever (MHF), Crimean-Congo hemorrhagic fever (CCHF) and Lassa fever. In particular, the causative agents of EHF, MHF, CCHF, and Lassa fever are Ebola, Marburg, CCHF, Lassa viruses, respectively, and regarded as biosafety level-4 pathogens because of their high virulence to humans. Recently, relatively large outbreaks of EHF and MHF have occurred in Africa, and areas of EHF- and MHF outbreaks seem to be expanding. Although outbreaks of VHF have not been reported in Japan, there is a possibility that the deadly hemorrhagic fever viruses would be introduced to Japan in future. Therefore, preparedness for possible future outbreaks of VHF is necessary in areas without VHF outbreaks. PMID- 16363690 TI - [Epidemiology of rabies virus and other lyssaviruses]. AB - Rabies is a zoonosis that infects domestic and wild animals through close contacts with saliva from infected animals. The annual number of deaths worldwide caused by rabies is estimated approximately 55,000 by World Health Organization (WHO). There has been no indigenous rabies case in Japan since 1957; however, there was only one imported case, a traveler who was bitten by a stray dog in Nepal and died in 1970. Dogs in Asia and Africa remain the main reservoir and transmitter of rabies to humans. The others are mainly coyotes, foxes, jackals, mongooses, raccoons, skunks, wolves and bats. The efficacy of the current human and veterinary vaccines against emergent lyssaviruses should be evaluated because the newly discovered rabies-related viruses have been isolated from bats. PMID- 16363691 TI - [Clinical feature of human rabies]. AB - Rabies is one of the most typical zoonosis that has been well known since ancient ages. Although no rabies case has been reported since 1957 in Japan, there are many areas where rabies is yet endemic or epidemic. Usually men contract rabies through rabid animal bite. However, human-to-human transmission of rabies virus occurred through organ transplantations. Rabies causes fatal encephalitis in animals and humans and effective methods to treat rabies patients have not yet been available. The only means to escape rabies death is to receive the post exposure prophylaxis of rabies with rabies vaccine as soon after animal bite as possible. We should keep in mind that rabies is preventable but incurable. PMID- 16363692 TI - [Rabies contingency plan in Japan]. AB - In Japan, rabies has been culled out since 1957 thanks to the strong implementation of measures against rabies, such as vaccination of dogs, quarantine and control of wild dogs under the 'Rabies Prevention Law' enacted in 1950. Nevertheless one cannot deny the possibility of introduction of rabies into Japan in view of the recent increase in the international movements of people and animals. Should an outbreak of rabies be suspected now in Japan, the society would probably overreact due to a decreased awareness of risks and a lack of correct knowledge about this disease. Officials of the government and the municipalities, veterinarians and doctors should exchange correct information on rabies and on prevention control and raise their awareness, while providing also information to the public on a timely basis. Besides it is needless to say that it is important to set up a crisis management system allowing a quick and adequate response in case of an outbreak of rabies and to continue to implement appropriate prevention measures in normal times. PMID- 16363693 TI - [Infectious disease of simian herpes B virus]. AB - The epidemiology and the method of diagnosis were elucidated for simian herpes B virus (SHBV) infection. It is important that usefulness was demonstrated for the methods of DNA diagnosis having high sensitivity and specificity for SHBV and HSV 1, 2 types, and of detectable serological diagnosis for each specific antibody. The methods allowed the final diagnosis of the human infection due to the reactivation of latent SHBV and the HSV infection from human to monkey. These results would be able to become important and fundamental knowledge for the sero epidemiological analysis of the infectious stile. PMID- 16363694 TI - [Hantavirus infection]. AB - Hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) are rodent borne viral zoonosis caused by hantavirus infection. HFRS and HPS cause epidemic in Eurasia continent and North and South American continent, respectively. Various species of wild rodents play as a reservoir animal in nature. Laboratory rat derived HFRS epidemic has also been reported in several countries including Japan. Recently, HFRS and HPS were classified to category 4 diseases in the Japanese law in which patient must be reported to municipality office. Since rat has widely been used in many institutions, prevention and control of the animal infection are quite important for both occupational hazard and research accomplishment point of view. PMID- 16363695 TI - [Tick-borne encephalitis]. AB - Tick-borne encephalitis (TBE) is viral zoonosis transmitted by the infected ticks harboring TBE virus. Mortality rate reaches 30% of in some cases. More than 10,000 patients were reported annually in Europe and Far-East Asia. In 1993, the first case of TBE was reported in Japan, and TBE virus was isolated, which showed TBE virus is endemic in Japan. PMID- 16363696 TI - [Rift Valley fever]. AB - Rift Valley fever is transmitted by mosquito bites. The causative agent was isolated in 1931 from an infected sheep in Kenya's Rift Valley. In east Africa, outbreaks usually occur every 5 to 10 years, probably due to movement of the inter-tropical convergence zone. The many shallow depressions, "dambos" in east and southern Africa, filled with water during the rainy season are the main habitat for mosquito larva. Rift Valley fever was confined to the South of the Sahara until 1977 when a big outbreak occurred in Egypt. One of the factors believed responsible for the outbreak was the abundant water supply from canals of the newly constructed (Aswan? ) dam. PMID- 16363697 TI - [Prion disease as infectious disease transmissible from animals to human]. AB - Prion diseases such as bovine spongiform encephalopathy (BSE) have been recognized as zoonosis since the existence of variant Creutzfeldt-Jakob disease (vCJD) was reported in 1996. After then, BSE became a serious social problem all over the world. The incidence of BSE in EU and UK appears declining, and the vCJD incidence also shows a tendency to decrease. On the contrary, fears for the spread of BSE became actual problems: BSE occurrence outside of EU, introduction of BSE to other ruminants, and transmission of vCJD by blood transfusion. To prevent further spread of animal prion diseases and to reduce their risk to human being, active surveillance for animal prion diseases, removal of specified risk materials from food and feed chains, and effective feed regulation for livestock should be important. For the disclosure and elimination of prion-contaminated blood, materials for medical and pharmaceutical products, it is required to improve the sensitivity of prion detection methods. Furthermore, establishment of the therapeutics for human prion diseases is urgent problem. PMID- 16363698 TI - [Hepatitis E]. AB - Hepatitis E is an important public health disease in developing countries where sanitary conditions are not well established. In developed countries, it is sporadic and mainly "imported". The causative agent of hepatitis E, hepatitis E virus (HEV), is a single-stranded positive-sense RNA virus, that belongs to the genus Hepevirus in the family Hepeviridae. Recent development of specific HEV RNA and HEV antibody detection revealed cases of indigenous HEV infection. Many cases are associated with uptake of raw meat of HEV-infected animals. Although the natural life cycle of HEV is not known, new aspects of HEV infection, zoonosis, is emerged. PMID- 16363699 TI - [Liver regenerative therapy using glycoside-modified bone marrow]. AB - Recent studies have reported that bone marrow cells (BMCs) have the ability to generate functional hepatocytes. However, the efficiency with which BMC transplantation generates functional hepatocytes is rather low. We assumed that if BMCs accumulated directly in liver, the functional BMC-derived hepatocytes should increase more efficiently. An attempt was made to increase the accumulation of BMCs directly in liver through the interaction between hepatic asialoglycoprotein receptor (ASGPR) and galactose-exposing BMCs. Galactose exposing BMCs that expressed green fluorescent protein (GFP) were injected into Long-Evans-Cinnamon (LEC) rats, a Wilson's disease (WD) model, via the tail vein. The WD is an autosomal-recessive disorder characterized by impaired biliary copper excretion and copper toxicosis, all due to mutations in the atp7b gene. At 5 months after transplantation, GFP-expressing hepatocyte nodules accounted for 2.4% of total liver mass, and the normal ceruloplasmin was detectable in the sera of these LEC rats. These findings suggest that the functional BMC-derived hepatocytes can be generated and the new genes derived from BMCs, such as ATP7B and GFP, can be transferred to LEC rats by the direct accumulation of BMCs in liver without hematopoietic reconstitution in need of preparative lethal irradiation. PMID- 16363700 TI - [Significance of insulin secretion pattern lectured by "glinides" in the treatment of postprandial hyperglycemia]. AB - The mechanisms by which postprandial hyperglycemia is elicited were discussed through therapies of type 2 diabetes using "glinides". It has been believed that the earliest determinant of progression to type 2 diabetes is a loss of early insulin secretion, a defect which results in postprandial hyperglycemia and is often believed to reflect insulin resistance. To prove that, we improved insulin secretion pattern without increase of total amount of insulin secretion using glinide and assessed glucose response. Glinide which selectively enhances early meal-induced insulin secretion improved postprandial hyperglycemia, could provide a valuable treatment option in the prevention and treatment of type 2 diabetes. PMID- 16363701 TI - [Rapid diagnosis of donor infection and renal allograft contamination using a polymerase chain reaction technique and rapid shaking culture]. AB - Post transplant infection is one of the serious complications of the organ recipients. We detected the donor infections and allograft contaminations in a limited period of time by polymerase chain reaction (PCR) and rapid shaking culture (RSC). The pre-procurement blood from 86 possible renal donors as well as the preservation solution (PS) and renal pelvic urine (PU) from 158 grafts were examined in order to detect highly virulent organisms such as methicillin resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and fungi. The average age of donors was 48.8 years old. The average period between the admission and brain death was 4.9 days, and the average period between the brain death and cardiac arrest was 3.6 days. The inflammatory indices such as body temperature, white blood cell count and C-reactive protein increased to 37.9 +/- 1.1 degree C, 12,600 +/- 6300/microl, and 20.2 +/- 11.6 mg/dl, respectively. Following PCR and RSC, procurement operations of the three possible donors were cancelled because of systemic bacterial infections by MRSA or Bacteroides fragilis. Six out of 158 grafts were discarded due to the diagnoses of MRSA or Candida albicans in PS and/or PU. All the other 148 grafts were transplanted. Following transplant, no single infectious complication transmitted by the graft was noted. We conclude that PCR combined with RSC is very accurate and useful for detecting donor infections and allograft contaminations, which may cause severe complications in the recipients. PMID- 16363702 TI - [Clinical analysis of bladder cancer patients undergoing radical cystectomy]. AB - A retrospective clinicopathological study was performed on 96 bladder cancer patients who underwent radical cystectomy and pelvic lymphadenectomy at Aichi Cancer Center between April 1994 and July 2004. The follow-up period ranged from 4 to 109 months, with a mean of 39.1 months. The overall cancer-specific survival rates at 3, 5 and 7 years were 81.7%, 71.2% and 71.2%, respectively. The 5-year survival rate according to pathological stage was 97.3% for pT1 or lower, 66.4% for pT2, 47.6% for pT3, and 25.4% for pT4. Univariate analysis demonstrated the prognostic significance of pathological stage (p < 0.001), lymph node metastasis (p = 0.001), venous invasion (p < 0.001), lymphatic invasion (p = 0.010), preoperative creatinine clearance (p = 0.001) and preoperative hydronephrosis (p < 0.001). Multivariate analysis revealed strong independent correlations of venous invasion (p = 0.040) and preoperative hydronephrosis (p = 0.016) with prognosis. PMID- 16363703 TI - [A stage-dependent follow-up strategy after radical cystectomy for bladder carcinoma]. AB - With the aim of developing stage-dependent follow-up strategy after radical cystectomy for bladder carcinoma, the records of 111 patients with bladder carcinoma who underwent radical cystectomy during the period between 1986 and 2003, were reviewed for the date and site of recurrence. Intrapelvic recurrence developed in 3 out of 56 patients with pT1> or =, 1 of 22 with pT2 and 6 out of 33 with pT3< or = at a median of 34 (range 32-58), 28, 8 (4-51) months, respectively. Extrapelvic recurrence developed in 20 patients with pT1> or =, 4 with pT2 and 14 with pT3< or = at a median of 43 (20-66), 15.5 (13-20), 8 (2-46) months, respectively. Recurrence developed earlier and more frequently in patients with pT3< or = and pT2 than those with pT1> or =. A stage-specific approach to tumor surveillance after radical cystectomy for bladder carcinoma, taking into consideration the risk of recurrence, represents a new approach for efficiently detecting recurrence and reducing medical costs. Our results offer the possibility of a new stage-specific approach to tumor surveillance after radical cystectomy for bladder carcinoma, for efficiently detecting recurrence and reducing medical costs, taking into consideration the risk of recurrence. PMID- 16363704 TI - [Urachal anomalies and tumor: clinical investigation of 14 cases]. AB - Diseases associated with persistent urachus are relatively rare. During the past 9-year period, there have been 14 patients with urachal disease consisting of 10 with urachal abscess and 4 with urachal cancer. The 10 patients with urachal abscess consisted of 7 males and 3 females aged 19-77 years (mean, 46 years). The 4 patients with urachal cancer consisted of 2 males and 2 females aged 48-81 years (mean, 57 years). As symptoms, lower abdominal pain was frequently observed in the patients with persistent urachus with abscess and gross hematuria in those with urachal cancer. Echo and magnetic resonance imaging (MRI) were useful for visualizing the lesion. Computed tomogtaphic (CT) scanning could not visualize the lesion in 2 patients. Nine patients underwent MRI, which visualized the lesion in all of them. As urachal abscess, an umbilical fistula was observed in 3 patients, urachal cyst in 4, and urachal diverticulum in 1. The preoperative diagnosis was urachal cancer in 6 patients, and pathological examination showed 4 patients with adenocarcinoma, 1 with inflammatory granuloma, and 1 with pseudosarcoma. Urachal abscess was treated by resection of the abscess in 6 patients, transurethral resection in 1, and resection of the umbilicus and urachus and total cystectomy in the other. Of the patients with urachal cancer, 1 underwent total cystectomy and the other 3 underwent total urachal resection and partial cystectomy. In 2 patients with persistent urachus with abscess, the differentiation between abscess and malignant tumor was difficult. PMID- 16363705 TI - [Clinical experience of streptococcal balanoposthitis in 47 healthy adult males]. AB - Between January 2001 and December 2003, 189 adult patients with balanoposthitis were treated in my clinic. Swab culture from the affected lesion detected Streptococcus pyogens in 47 cases (PYO group), other bacterial species including Candida albicans in 93 (B group), no bacterial growth in 27 (NB group) and swab culture was not done because clinical symptom was trivial in 22 (N group). The PYO group had a significantly higher prevalence of purulent discharge (68.1%) and local pain (38.3%), compared to the B group (25.8% and 21.5%, respectively) or the NB group (33.3% and 11.1%, respectively). Phimosis was absent in 7 cases in the PYO group. The route of infection in the PYO group was considered to be predominantly via sexual contact (PYO group 78.7%, B group 52.7%, NB group 59.3%), especially through fellatio by commercial sex worker for the majority of the PYO patients. The latent period (from sexual contact to the onset of symptoms) was 3 days or less in 40.5% and 4 to 7 days in 35.1% in the PYO group, whereas it was more than a week or not remembered in the majority of the B and NB groups, disapproving the causal relationship with sexual contact in such cases. Treatment with tosufloxacin tosilate or amoxicillin was effective in most cases of streptococcal balanoposthitis. Our results suggest that Streptococcus pyogens is one of the causative organisms of sexually transmitted balanoposthitis. PMID- 16363706 TI - Renal transcatheter arterial embolization for autosomal dominant polycystic kidney disease: report of two cases. AB - We report the results of renal transcatheter arterial embolization (renal-TAE) in two patients with autosomal dominant polycystic kidney disease (ADPKD) treated with hemodialysis (HD). A 73-year-old man and a 65-year-old woman on HD visited our department complaining of abdominal fullness, abdominal pain and appetite loss. Abdominal computerized tomography (CT) revealed polycystic kidneys. Both patients underwent renal-TAE. Approximately 1 month later, the symptoms had improved and CT showed decreased cyst volume. These results suggest that renal TAE is a useful therapy for patients with ADPKD. PMID- 16363707 TI - [Oncocytoma associated with acquired cystic disease of kidney (ACDK): a case report]. AB - A case of renal oncocytoma associated with acquired cystic disease of kidney (ACDK) in a 56-year-old man is reported. He had received hemodialysis for 15 years because of chronic renal failure. Computed tomography (CT) was performed because of distention in the upper abdomen, revealing a right renal tumor. He underwent laparoscopic right nephrectomy, and was diagnosed with renal oncocytoma. There have been reported 8 cases of renal oncocytoma in hemodialysis patients, and our case was the third one associated with ACDK in the literature. PMID- 16363708 TI - [A case of primary retroperitoneal mucinous cystadenocarcinoma]. AB - A 41-year-old Japanese woman was admitted to our hospital for right abdominal mass. Various examinations revealed a retroperitoneal lymphangioma. Tumor resection was performed and pathological diagnosis was a mucinous cystadenocarcinoma. It seems that screening with the tumor markers will be helpful for the diagnosis, because it is very difficult to diagnose a retroperitoneal mucinous cystadenocarcinoma before the operation. PMID- 16363709 TI - [Invasive bladder cancer recurrenced 5 years after complete response status by chemotherapy and radiotherapy: a case report]. AB - A 72-year-old man had undergone trasucethral resection of bladder tumor (TUR-Bt) three times from 1990 to 1991 and he had been lost to follow with no recurrence from 1996, came to our hospital complaining of asymptomatic macrohematuria in May 1999. A bladder tumor existed around the right ureteral orifice with right hydronephrosis. MRI and TUR-Bt revealed that the cancer was transitional cell carcinoma (TCC) > small cell carcinoma, G3, pT3b. Because the patient insisted on bladder preservation, intra arterial chemotherapy with cisplatinum (CDDP) and epirubicin (EPI-adr) followed by radiotherapy with CDDP was performed. The treatment resulted in a clinical complete response (CR), and the bladder was preserved. In January 2004, an invasive bladder cancer recurred at the left lateral wall. This time, neoadjuvant intra-arterial chemotherapy with CDDP and EPI-adr, followed by radical cystectomy was performed. Histologically, the recurrent bladder cancer was TCC, G3, pT3b. PMID- 16363710 TI - [Inguinal lymph node metastasis of bladder carcinoma ten years after cystourethrectomy: a case report]. AB - A 79-year-old man had undergone radical cystourethrectomy for bladder carcinoma in January, 1989. Pathological report was Small cell carcinoma (SCC) >> transitional cell carcinoma (TCC), G2 > G3, pT4 (prostate), ew (-). Ten years later, follow-up computed tomography (CT) revealed swollen left inguinal lymph node in October 1998 and lymph node dissection was performed in January, 2000. The pathological report showed TCC, G2. Left inguinal lymph node metastases appeared again in January, 2001. Chest X-ray films showed multiple lung metastases in March, 2001. Three couses of MVAC (methotrexate, vinblastine, doxorubicin cisplatinum) chemotherapy had been performed since September, 2001 but were in effective. Papillary tumor was observed at external urethral meatus in September, 2002 and the biopsied specimens showed TCC, G1 > G2, pathologically. Finally he died of respiratory insufficiency in January, 2003. It is suggested that the recurrent TCC tumor in the urethral remnants might metastasize into the inguinal lymph nodes. PMID- 16363711 TI - [Ejaculatory disorder by alpha-1 adrenoceptor antagonist in patients with benign prostatic hyperplasia; retrospective comparison between naftopidil and tamsulosin]. AB - We cross-sectionally studied the incidence and impact on quality of life of ejaculatory disorders caused by alpha-1 adrenoceptor antagonists (tamsulosin or naftopidil) in patients with benign prostatic hyperplasia (BPH). By questionnaire, we queried 88 clinical BPH patients concerning ejaculatory disorders, who were treated with tamsulosin or naftopidil between February 1999 and August 2003. We investigated the difference in the incidence and types of disorders between the two drugs. Of the 88 patients, 63 (71.6%) had sexual activities during the treatment. The incidence of ejaculatory disorders was significantly higher in the tamsulosin group (30.0%) than in the naftopidil group (3.0%). Eighty percent of patients having the disorders noticed the absence or reduced volume of semen although they experienced orgasms. The median quality of life index concerning the disorder was rated as 4, "mostly dissatisfied." Our results indicated that ejaculatory disorders occurred more frequently in the tamsulosin group. We should inform patients about this adverse effect caused by alpha-1 adrenoceptor antagonists prior to treatment. PMID- 16363712 TI - [Inflammatory pseudotumor of the prostate: a case report]. AB - We report a rare case of inflammatory pseudotumor of the prostate. A 42-year-old man with a history of hematospermia and chronic prostatitis presented with difficulty in voiding. Cystoscopy demonstrated a large non-papillary tumor occupying the prostatic urethra along with two bladder stones. Magnetic resonance imaging (MRI) demonstrated a 7-cm prostatic mass protruding toward the bladder and the rectum. Transrectal biopsy of the prostate demonstrated a fibrous lesion containing inflammatory cells without evidence of malignancy. We performed transurethral resection of the prostatic lesion to release the bladder outlet obstruction, followed by cystolithotripsy. Histopathological examination of the surgical specimen demonstrated a benign fibromuscular lesion with spindle cell proliferation, leading to a diagnosis of inflammatory pseudotumor. Postoperatively, the patient voided normally without any signs of recurrence on follow-up at five months. Inflammatory pseudotumor is an unusual benign lesion of unknown etiology. Only 10 previous cases of the disease involving the prostate have been reported in English and Japanese literature. Prostatic involvement of inflammatory pseudotumor may show a presentation similar to malignant prostatic sarcoma. Thus, accurate identification of this benign process is important in order to avoid unnecessary radical surgery. PMID- 16363713 TI - [Prostate cancer with penile metastasis: a case report]. AB - A 77-year-old man presented with complaints of dysuria, nocturia and painless nodule on his penis. Laboratory examination revealed elevated serum prostate specific antigen (PSA) and CA19-9. Pathological examinations on prostate and penile biopsy specimens revealed prostate adenocarcinoma with penile metastasis. The patient was diagnosed as having prostate cancer stage D2 (T4N1M1) with bone, lymph node and penile metastases. There was no response to initial hormonal therapy with the surgical castration and diethylstilbestrol. However, decrease of the tumor size, as well as PSA and CA19-9 values were achieved after the combined chemotherapy with Estramustine, Paclitaxel and Carboplatin. PMID- 16363714 TI - [A case of spindle cell carcinoma of the penis]. AB - Spindle cell carcinoma of the penis is extremely rare. The present case report is the nineth case in the world literature. A 71-year-old man was referred to our institution for examination of a penile tumor that showed non-specific granuloma histopathologically. We performed biopsy and total tumor resection. The histopathological finding showed squamous cell carcinoma with spindle type cells constituting the majority of the tumor, and the switching over from squamous cell carcinoma to spindle cell carcinoma was clearly visible in the histopathological examination. Therefore, we diagnosed this as spindle cell carcinoma of penis and performed partial penectomy with inguinal lymph node dissection. Spindle cell carcinoma is an uncommon variant of squamous cell carcinoma. We report the histopathological findings including the data of p53 immunohistochemical study, and reviewed the eight previous reports of spindle cell carcinoma of the penis. PMID- 16363715 TI - Use of an aortic connector system for vein grafting: intermediate outcomes. AB - OBJECTIVE: The aortic connector system was used to minimize cerebrovascular complications when performing the proximal anastomosis of vein grafts during coronary artery bypass grafting (CABG). The goal of this study was to investigate the intermediate outcomes of patients undergoing CABG with the aortic connector system. METHODS: The aortic connector was used on nine patients undergoing CABG between November 2002 and July 2003. Intermediate outcomes of the patients were examined, and the results of coronary angiography, which were performed before patient discharge and at least 6 months after discharge, were evaluated. RESULTS: There were no operative deaths or cerebrovascular accidents. One patient died 9 months after discharge, one patient had angina, and the remaining seven patients were asymptomatic. When evaluating the results of angiography performed before patient discharge, two of the 21 distal vein graft anastomoses were occluded (patency rate, 90.5%), but there was no stenosis or occlusion at the proximal anastomoses sites that were performed using the aortic connector. When evaluating the results of the second angiography performed after patient discharge, four of the eight proximal anastomoses were patent, one was completely occluded, two had 90% stenosis and one had 75% stenosis. Further, four of the 18 distal anastomoses were occluded (patency rate, 77.8%). There was no significant difference in graft flow or device size when comparing patients with patent vein grafts and those with stenotic or occluded vein grafts. CONCLUSION: Intermediate outcomes of vein grafting using the aortic connector were suboptimal. Long-term outcome data are forthcoming. PMID- 16363716 TI - Mid-term results of open aortic valvotomy for infants with critical aortic stenosis: seven-year experience including delayed Ross strategy. AB - OBJECTIVES: The purpose of this study is to provide short- and mid-term results of open aortic valvotomy (OAV) for patients with critical aortic stenosis (AS). METHODS: Between December 1993 and June 1996, 6 patients with critical AS underwent an OAV in our unit. Their ages and body weights at operation ranged from 1 to 65 days (median age, 9 days) and from 2.4 to 5.7 kg (median weight, 3.3 kg), respectively. Peak pressure gradient and diameter of the aortic valve ranged from 25 to 111 mmHg (mean value, 79 mmHg) and from 4.6 to 7.5 mm (mean diameter, 6.1 mm), respectively. OAV comprised the valvular commissurotomy and excision of the myxomatous nodules with cardiopulmonary bypass. RESULTS: No early or late death occurred. Mean peak pressure gradient across the aortic valve was reduced to 33 mmHg (from 15 to 44 mmHg) with no aortic insufficiency in 2 patients and trivial insufficiency in 4. During the follow-up period of 6 to 9 years, 3 out of 6 patients required no reintervention. The other 3 patients required repeated valvotomy for recurrent stenosis within 0.2 to 1.3 years after the operation. Of these, 2 patients required the Ross procedure at 7 years of age or older, and another at 6 years of age awaits the Ross procedure. CONCLUSION: OAV for critical AS was effective without causing mortality or significant aortic insufficiency. Our current strategy comprising the initial OAV and "delayed Ross procedure" for recurrent stenosis with or without insufficiency is a promising therapeutic option for infants with critical AS. PMID- 16363717 TI - Intramyocardial foreign body: sewing needle with the uncommon clinical feature of constrictive pericarditis. AB - A rare case of intracardiac injury with a sewing needle causing constrictive pericarditis is reported. A 72-year-old woman was admitted with edema of the face and legs. A chest radiogram showed a needle-shaped metallic image overlying the cardiac silhouette. Computed tomography revealed the needle was located in the left ventricular myocardium, and also showed a thickened pericardium. Surgical removal of the needle and pericardectomy were performed simultaneously. PMID- 16363718 TI - Successful application of pulmonary arterial patch for coronary angioplasty late after arterial switch operation. AB - Pulmonary artery is infrequently utilized as a material for coronary patch angioplasty in children. We applied a pulmonary arterial patch for coronary angioplasty on an 8-year-old boy with total occlusion of the left main coronary artery late after an arterial switch operation. The pulmonary arterial patch was easy to handle and the immediate result after the operation was satisfactory. PMID- 16363719 TI - Spontaneous pneumothorax and a coexistent azygos lobe. AB - We present a primary spontaneous pneumothorax involving a possible diagnostic and surgical pitfall. A 25-year-old man with a previously identified azygos lobe was admitted to our hospital because of right primary spontaneous pneumothorax. A preoperative chest X-ray at the onset of the pneumothorax as well as after resolution by tube drainage showed an azygos fissure but no azygos vein. Chest computed tomography revealed the dislocated azygos vein external to the azygos fissure on the mediastinal side of the reexpanded upper lobe. Thoracoscopic bullectomies were uneventfully performed. The azygos arch was found dangling in the free inferior border of the mesoazygos. An azygos vein located in this position can be a potential surgical hazard especially in video-assisted thoracic surgery. This case suggests that the presence of an azygos lobe can be missed on chest X-ray when a pneumothorax occurs in a patient with an azygos lobe. PMID- 16363720 TI - Dissecting aortic aneurysm in a renal transplant recipient. AB - There have been few reports of surgical repair of acute aortic dissection in renal transplant recipients. The incidence, operative risk, or perioperative management of aortic dissection with functioning allografts remains unknown. Herein we report our experience in successful treatment of type I dissecting aortic aneurysm in a renal transplant patient. A 35-year-old man was admitted to our hospital complaining of severe chest pain. He had undergone a living renal transplant from his mother for chronic renal failure caused by immunoglobulin A nephropathy 11 years prior to admission. An immunosuppressive regimen had been maintained continuously. Preoperative chest computed tomography demonstrated a thoracic dissecting aortic aneurysm (DeBakey classification type I). An emergent graft replacement for the ascending aorta was placed under circulatory arrest. Although continuous hemodiafiltration was required postoperatively because of deteriorated renal function, he recovered uneventfully and his renal function returned to preoperative values. He was discharged on postoperative day 26 without any complications. PMID- 16363721 TI - Pulmonary blastoma. AB - We describe a resected pulmonary blastoma in an 84-year-old male, the oldest of previously reported patients. A chest X-ray showed a 2 cm-sized abnormal shadow in the left lung field. Five months later computed tomography demonstrated a well demarcated heterogenous mass, measuring 12 cm in diameter, in the left lower lobe of the lung. This mass was diagnosed as a carcinoma using echo-guided percutaneous biopsy. The patient underwent a left lower lobectomy. The resected specimen revealed the tumor to be a pulmonary blastoma arising from lung tissue. Preoperative diagnosis of pulmonary blastoma is extremely difficult because of the histological heterogeneity of tumor. Since it has been noted that pulmonary blastoma rapidly progresses in a short period of time, surgical treatment should be undertaken as quickly as possible when such a tumor is suspected. PMID- 16363722 TI - Surgical treatment of a retrosternal pseudoaneurysm arising from the posterior aortic root. AB - We report a case of a 60-year-old male with a retrosternal pseudoaneurysm arising from the posterior aortic root. He had undergone replacement of the aortic root and ascending aorta for an acute aortic dissection. His postoperative course was complicated with composite graft infection. During a redo surgery, femoro-femoral bypass was established prior to sternotomy via the right femoral artery and vein, and ten French cannulas were directly inserted into both carotid arteries through separate skin incisions for brain perfusion. When the sternum was divided, an occlusion balloon catheter introduced through the left femoral artery was inflated to maintain somatic perfusion without compromising clear vision of the operative field. Combined procedures with direct cannulation into both common carotid arteries and balloon occlusion of the distal aortic arch allowed us to safely perform replacement of the aortic root and ascending aorta in the patient with a retrosternal pseudoaneurysm. PMID- 16363723 TI - Aortobronchial fistula resulting from a mycotic pseudoaneurysm after treatment of an aortoesophageal fistula due to a thoracic aortic aneurysm. AB - A 58-year-old man was admitted for an aortoesophageal fistula (AEF) resulting from a thoracic aortic aneurysm. He underwent immediate in-situ prosthetic graft replacement, primary esophageal repair and wrapping of the aneurysm. Postoperative upper gastrointestinal endoscopy and computerized tomography (CT) findings were unremarkable. He was discharged on postoperative day (POD) 25. Three months after surgery, he was readmitted with complaints of worsening cough and hemoptysis. CT showed a thrombosed aneurysm adjacent to the left bronchus. Aortobronchial fistula due to mycotic pseudoaneurysm was suspected. The patient underwent immediate resection of the infected graft and prosthetic graft replacement positioned to avoid the infected area. The graft was wrapped with omentum. On POD 7, pleural empyema developed, and esophagography revealed a residual leak. Staged reconstruction of the esophagus was performed successfully. We conclude that even if the fistulous opening is small, simultaneous esophageal resection should be performed during the initial treatment of AEF. PMID- 16363724 TI - Primary cardiac angiosarcoma resection combined with right coronary artery bypass grafting. AB - A 41-year-old woman presented with symptoms of cardiac tamponade. Diagnosis of a primary malignant tumor in the right atrium without distant metastasis was made by echocardiography, computed tomography and coronary angiography. Aggressive surgery consisting of resection of the tumor including the right atrial wall together with the right coronary artery and sinus node was performed. This was followed by coronary artery bypass grafting with myocardial electrode implantation for reconstruction of the right atrial wall using an equine pericardium. Final diagnosis was of angiosarcoma by pathological examination. Consequently, the patient underwent additional radiotherapy and was discharged. She is currently in good health with no recurrence 1.5 years after surgery. PMID- 16363725 TI - [Current problems surrounding pneumonia]. PMID- 16363726 TI - [Bacterial pneumonia (with special reference to Streptococcus pneumoniae pneumonia)]. PMID- 16363727 TI - [Mycoplasma pneumonia]. PMID- 16363728 TI - [Chlamydia pneumonia]. PMID- 16363729 TI - [Legionella pneumonia]. PMID- 16363730 TI - [Pneumonia associated with influenza]. PMID- 16363731 TI - [Pulmonary tuberculosis]. PMID- 16363732 TI - [Infection by non-tuberculous acid-fast bacteria]. PMID- 16363733 TI - [Mycotic pneumonia]. PMID- 16363734 TI - [Parasitic lung disease]. PMID- 16363735 TI - [Q fever]. PMID- 16363736 TI - [Pneumonia among the aged]. PMID- 16363738 TI - [Ventilator-associated pneumonia]. PMID- 16363737 TI - [Nosocomial pneumonia]. PMID- 16363739 TI - [Pneumonia as an opportunistic infection]. PMID- 16363740 TI - [Guidelines for management of pneumonia]. PMID- 16363741 TI - [Pneumonia: progress in its diagnosis and treatment. Discussion]. PMID- 16363742 TI - [Acute disseminated encephalomyelitis (ADEM) with Elsberg syndrome: clinical report of two cases]. PMID- 16363743 TI - [Spontaneous intracranial hypotesion: demonstration of cerebrospinal fluid leakage by magnetic resonance myelography]. PMID- 16363744 TI - [32 year old male case of paraneoplastic non-herpetic limbic encephalitis with thymoma]. PMID- 16363745 TI - [HHV-6 encephalitis after unrelated cord blood transplantation in a patient with acute lymphoblastic leukemia]. PMID- 16363746 TI - [Approval of new antibacterial agents and postmarketing survey: important information for practitioners]. PMID- 16363747 TI - [Prevention of accidental falls by the aged]. PMID- 16363748 TI - [Familial amyloid polyneuropathy: variability of clinical presentations and therapeutic effects of liver transplantation]. PMID- 16363749 TI - [Metabolic syndrome and an intra-cellular glucocorticoid activating enzyme, 11 beta-HSD1]. PMID- 16363750 TI - [The current status of drug sensitivity tests for chemotherapy of lung cancer]. PMID- 16363751 TI - Resident job satisfaction and quality of life before and after work hour reform. AB - OBJECTIVE: To assess the effect of limiting work hours for obstetrics/gynecology residents on working conditions, job satisfaction and quality of life. STUDY DESIGN: A survey assessing hours worked, working conditions, job satisfaction and quality of life during residency was sent to 10 current residents and 25 residency graduates. Council on Resident Education in Obstetrics and Gynecology scores were also examined. RESULTS: The average hours worked per week dropped (89 to 82.5, p < 0.02). Hours of sleep obtained on call decreased (3 to 2, p < 0.008). Overall, 25% of subjects reported that their clinical judgment was adversely affected by a 24-hour shift and 64% by a 36-hour shift. Seven percent of subjects reported that a 24-hour shift affected their surgical performance, while 54% reported that their surgical skills were adversely affected by a 36 hour shift. Satisfaction with work and Council on Resident Education in Obstetrics and Gynecology scores did not vary significantly. There was an improvement in residents' satisfaction with personal life (2.8 to 1.8 average rating, p < 0.05). CONCLUSIONS: Accreditation Council of Graduate Medical Education reforms improved resident's satisfaction with their personal lives but not CREOG scores or satisfaction with work. PMID- 16363752 TI - Abruptio placentae associated with misoprostol use in women with preeclampsia. AB - OBJECTIVE: To evaluate complications associated with cervical ripening with vaginal administration of misoprostol and dinoprostone vaginal inserts in women with preeclampsia. preeclampsia. STUDY DESIGN: Retrospective study of patients with preeclampsia undergoing cervical ripening with vaginal misoprostol and dinoprostone vaginal inserts prior to labor induction. RESULTS: Among 203 patients with preeclampsia undergoing cervical ripening prior to induction, 95 received vaginal misoprostol, and 108 received dinoprostone. The incidence of uterine hyperstimulation requiring medical therapy and the need for emergency cesarean section due tofetal heart rate abnormalities were significantly higher among patients receiving misoprostol (22.1% versus 12.0%, p = 0.04, and 17.9% versus 8.3%, p = 0.03, respectively). The overall incidence of abruptio placentae was 7.4%, with a significantly higher incidence among those receiving misoprostol as compared to dinoprostone (13.7% versus 1.9%, p = 0.001). CONCLUSION: Among patients with preeclampsia undergoing cervical ripening prior to labor induction, there is a higher incidence of acute intrapartum complications (uterine hyperstimulation, cesarean section for fetal heart rate abnormalities and abruptio placentae) with vaginal misoprostol, as compared to dinoprostone, vaginal insert. PMID- 16363753 TI - Incidence of significant adhesions at repeat cesarean section and the relationship to method of prior peritoneal closure. AB - OBJECTIVE: To examine the effect of peritoneal closure at prior cesarean section on the occurrence of abdomino-pelvic adhesions at subsequent cesarean section. STUDY DESIGN: An observational chart review study was performed involving 191 cases of repeat cesarean section occurring from January 2001 through July 30, 2002. An adhesion score was created according to the following: a grade 1 score was assigned when no adhesions were present, a grade 2 when adhesions were present but no surgery required, a grade 3 when adhesiolysis was required and there was inability to exteriorize the uterus, and a grade 4 when adhesiolysis was required prior to delivery. Scores were assigned and tabulated as a result of the review of the operative records of 191 cases. When available, the operative record of prior cesarean section was examined to determine if visceral and parietal peritoneal closure had been performed at the prior operation. RESULTS: During the year and a half for which data were examined, significant abdominopelvic adhesions (grades 3 and 4) were encountered 21% of the time (40 of 191) at repeat cesarean section. Prior peritoneal closure resulted in significant adhesions in 1 of 18 patients compared to 17 of 40 patients when no prior peritoneal closure was recorded (Fisher's exact probability = 0.003). CONCLUSION: While preliminary, in the absence of any substantive benefit or published data regarding adhesionformation in cesarean section that contradict this finding, the practice of nonclosure of visceral and parietal peritoneum at cesarean section should be questioned. PMID- 16363754 TI - Cocaine use during pregnancy and the failure of vaginal birth after cesarean. AB - OBJECTIVE: To evaluate the failure rate of a trial of vaginal birth in women with singleton pregnancies and 1 prior cesarean section who reported using cocaine during the current pregnancy. STUDY DESIGN: The medical records of 9254 women who attempted a vaginal birth after cesarean (VBAC) at 16 community and tertiary care hospitals were retrospectively reviewed. VBAC was considered a failure if a woman who was initially considered suited to a trial of VBAC for whatever reason did not succeed in vaginal delivery and had to undergo a repeat cesarean section. We used stratified analysis as well as forward and backward logistic regression analysis to assess the relation between cocaine use during pregnancy and VBAC failure. RESULTS: Thirteen percent of patients who reported using cocaine had a significant 59% decrease in the odds for VBAC failure in the unadjusted analysis (OR = 0.41, 95% CI= 0.29-0.58) as compared to those who did not report using this substance. Nevertheless, after adjusting for other important predictors of failure and possible confounding variables, the use of cocaine in our final model showed no difference in the odds for VBAC failure (OR = 0.74, 95% CI = 0.49-1.13) as compared to the odds in women who did not report using the drug. CONCLUSION: We could not find any statistically significant difference in the odds for VBAC failure between patients who reported cocaine use during pregnancy and patients who reported those who did not. PMID- 16363755 TI - Sexual functioning after vaginal hysterectomy or transvaginal sacrospinous uterine suspension for uterine prolapse: a comparison. AB - OBJECTIVE: To examine changes in sexuality after total vaginal hysterectomy (TVH) or transvaginal sacrospinous uterine suspension (SSUS)for uterine prolapse. STUDY DESIGN: One hundred fifty-eight women with moderate to severe uterine prolapse undergoing TVH (78) or SSUS (80) were included in a prospective study from January 2001 to June 2002. All women were <50 years old and sexually active within the last 6 months before surgical intervention. None had major medical disorders. Sexual functioning before and 6 months after surgery was examined via a face-to-face questionnaire. Sexual interest, sexual satisfaction, frequency of sexual intercourse and frequency of orgasm were measured using an analogue scale. RESULTS: Of women undergoing TVH, 5.1% had decreased sexual interest, and 21% had less frequent orgasms postoperatively. For women undergoing SSUS, 13% had decreased sexual interest, and 20% had less frequent orgasms postoperatively. Frequency of orgasm was the only parameter that changed significantly after surgery in the 2 groups. All women with less frequent orgasms said that they were afraid of wound disruption or disease recurrence and so refrained from vigorous or exciting sexual intercourse. About four-fifths of the women in both groups accepted or were satisfied with their sexuality after surgery. For women undergoing TVH, 2.6% had increased frequency of orgasm, and 5% had better overall sexual satisfaction postoperatively. For women undergoing SSUS, 10% had increased sexual satisfaction postoperatively. There was a 2.5% and 2.6% increase in postoperative sexual interest in the SSUS and TVH groups, respectively. The sexual functioning scores were not different before or after surgical intervention in either group. CONCLUSION: There is a decrease in thefrequency of orgasm after both TVH and SSUS. However, there is no significant difference in postoperative sexual functioning between women with and without preservation of the uterus in correcting uterine prolapse. PMID- 16363756 TI - Trisomies and other chromosome abnormalities detected after positive sonographic findings. AB - OBJECTIVE: To evaluate the rate of trisomies and other chromosome abnormalities after positive ultrasound findings in thefirst and second trimester of pregnancy. STUDY DESIGN: The study investigated chromosome abnormalities detected in cases with prior abnormal ultrasoundfindings. During a 10-year period there were 1907 invasive interventions carried out with the purpose of chromosome analysis. The intervention was genetic amniocentesis in 1619 cases and chorionic villus sampling in 288. RESULTS: Karyotyping revealed 103 cases (5.4%) of chromosome abnormalities. Abnormalities with subcutaneous edema were examined: abnormal karyotype was found in 20% of cases with nonimmune hydrops, 48.1% of cases with cystic hygroma and 53.8% of cases with nonimmune hydrops and cystic hygroma together, 8.3% of cases with nuchal edema in the first trimester and 5.5% in the second trimester. The incidence of chromosome abnormalities in cases of cerebral anomalies was 6.3% of cases with ventricular dilatation, 3.6% of cases with choroid plexus cysts and 15.9% of cases with other cranial anomalies. Regarding abnormalities of the heart, isolated echogenic intracardiac focus and ventricular septal defects were not associated with chromosome abnormality, but, in conjunction with other positive ultrasound findings, the incidence of chromosome abnormalities was 7.9% and 26.7%, respectively. Other anomalies of the heart and large blood vessels showed an abnormal karyotype incidence of 18.2%. In cases of unilateral pyelectasis unassociated with other anomalies, the incidence of chromosome abnormalities was 1%. In cases of bilateral pyelectasis or pyelectasis associated with other anomalies, the incidence was 3%. In terms of anomalies of the abdominal wall and abdomen, the incidence of association with chromosome abnormalities was 9.5% in cases of omphalocele, 11.8% in cases of duodenal atresia and 5.7% in cases of echogenic bowel. In cases of short femur and humerus the rate of abnormal karyotypes was 16%. CONCLUSION: Ultrasound plays an important role in prenatal diagnosis. In cases of positive ultrasound findings, karyotyping is reasonable. PMID- 16363757 TI - Chromosomal studies on 2 mL of celomic fluid obtained during the fifth week of development in the timed-pregnant baboon model. AB - OBJECTIVE: To determine if chromosomal studies could be performed using 2 mL of celomicfluid obtained during the fifth postfertilization week in pregnant baboons. MATERIALS AND METHODS: Nine ultrasound-guided celocenteses were performed. The initial 0.5 mL of celomic fluid was discarded to decrease maternal cell contamination. Approximately 2 mL of celomic fluid was then collected. The fluid was centrifuged and the supernatant removed to a final volume of 0.5 mL. The celomic fluid sample was placed in either a small plastic flaskette chamber slide with a mix of 0.5 mL celomic fluid, 1 mL of Amniomax, and 1 mL of usedfibroblast culture medium to spread on the entire surface (n=4), or a 3.5 x 1 cm plastic Petri dish with a 24 x 30-mm glass coverslip to keep the 0.5 mL celomic fluid mixed with 1 mL of Amniomax (Invitrogen, Carlsbad, California) within a 1 cm2 area (n=5). The medium was changed on day 5 and thereafter every second to third day. The cells were harvested when the number of cells appeared sufficient for chromosomal analysis. RESULTS: Standard chromosomal studies were possible in 5 of the 9 celomicfluid samples. Mean (+/-SD) celomic fluid volume used for culture was 1.85 +/- 0.3 mL. Mean (+/-SD) time to karyotype result was 18.8 +/- 1.8 days. CONCLUSION: The findings of this study suggest that there are living cells at 36-42 days of embryonic development in the extraembryonic celomic fluid of primates and that they can be cultured for chromosomal studies. However, significant improvements in understanding the biology of cells present at 5 weeks after fertilization in celomic fluid are needed to improve culture conditions. PMID- 16363758 TI - Effect of prostaglandin E2 for cervical priming on fetal hemodynamics. AB - OBJECTIVE: To assess the effect of the prostaglandin E2 (PGE2) on fetal vascular blood flow during cervical priming. STUDY DESIGN: In a prospective, observational study, 32 pregnant women > or =37 weeks of gestation requiring PGE2 for induction of labor were recruited. The resistance indices (RIs) of umbilical artery, middle cerebral artery, renal artery and ductus venosum were measured before, 2 and 4 hours after insertion of a PGE2 vaginal pessary. RESULTS: There were no significant changes in RIs of various fetal vessels during cervical priming with PGE2. CONCLUSION: There was no significant change in fetal hemodynamics after vaginal prostaglandin administration. This finding provides further evidence to support the safety of prostaglandin for normal fetuses. PMID- 16363760 TI - Nitric oxide inhibition of the proliferation of ovarian endometriotic stromal cells in vitro. AB - OBJECTIVE: To investigate the effects of nitric oxide on endometrial cell proliferation and the effects of peritoneal fluid from women with and without endometriosis on the production of nitric oxide and upon the nature of nitric oxide-induced changes. STUDY DESIGN: Ovarian endometriotic and endometrial stromal cells and peritoneal fluid were obtained from endometriosis patients and controls. Cell proliferation was determined by [3H]thymidine incorporation, and nitrite was measured using Griess reagent. RESULTS: Sodium nitroprusside (SNP), a nitric oxide donor, reduced the proliferation of endometriotic and endometrial cells in primary culture in a dose-dependent manner. SNP-induced production of nitric oxide and the inhibitory effect of nitric oxide on stromal cell proliferation were reduced by peritoneal fluid from women with endometriosis, although stromal cell proliferation was still inhibited by SNP in the presence of peritoneal fluid. However, this SNP-induced inhibition of cell proliferation was unaffected by interleukin-8, 17-beta, estradiol or transforming growth factor beta1. CONCLUSION: Nitric oxide inhibits the proliferation of endometrial stromal cells in vitro, and this inhibitory effect is abrogated by peritoneal fluid from women with endometriosis. PMID- 16363759 TI - Routine history as compared to audio computer-assisted self-interview for prenatal care history taking. AB - OBJECTIVE: To compare endorsement rates obtained with audio computer-assisted self-interview versus routine prenatal history. STUDY DESIGN: A crosssectional study compared items captured with the routine history to those captured with a computer interview (computer screen displaying and computer audio reading questions, with responses entered by touch screen). The subjects were women (n=174) presenting to a public hospital clinic for prenatal care. RESULTS: The prevalence of positive responses using the computer interview was significantly greater (p < 0.01) than with the routine history for induced abortion (16.8% versus 4.0%), lifetime smoking (12.8% versus 5.2%), intimate partner violence (10.0% versus 2.4%), ectopic pregnancy (5.2% versus 1.1%) and family history of mental retardation (6.7% versus 0.6%). Significant differences were not found for history of spontaneous abortion, hypertension, epilepsy, thyroid disease, smoking during pregnancy, gynecologic surgery, abnormal Pap test, neural tube defect or cystic fibrosis family history. However, in all cases, prevalence was equal or greater with the computer interview. CONCLUSION: Women were more likely to report sensitive and high-risk behavior, such as smoking history, intimate partner violence and elective abortion, with the computer interview. The computer interview displayed equal or increased patient reporting of positive responses and may therefore be an accurate method of obtaining an initial history. PMID- 16363761 TI - Asymptomatic uterine rupture of a cornual pregnancy in the third trimester: a case report. AB - BACKGROUND: Reports of cornual pregnancy persisting until fetal viability and of ultrasound diagnosis of asymptomatic uterine rupture are rare. CASE: A 24-year old woman, gravida 6, para 5, presented for initial ultrasound evaluation at 28 5/7 weeks' gestation after registering late for prenatal care. Her history included 2 prior cesarean deliveries. Obstetric ultrasound evaluation revealed a clinically silent uterine rupture with a live fetus. The site of rupture was suspected to be the prior uterine scar. After completion of a course of antenatal corticosteroids, the patient underwent exploratory laparotomy, abdominal delivery of a live, male neonate, hysterectomy and appendectomy. The patient experienced an unremarkable postoperative course and was discharged on postoperative day 4. The infant was discharged on hospital day 83 with chronic lung disease, stage 1 retinopathy of prematurity, atrial septal defect and a right clubfoot; there were no additional findings at the 4-month follow-up. Pathologic examination demonstrated cornual implantation with subsequent uterine rupture. CONCLUSION: This case demonstrates the importance of maintaining a suspicion for ectopic pregnancy at advanced gestational ages and for uterine rupture even in the absence of symptoms. The case also illustrates that fetal survival can occur in cornual pregnancy. PMID- 16363762 TI - Second-trimester tubal pregnancy: a case report. AB - BACKGROUND: Second-trimester tubal pregnancy is an infrequent diagnosis, as these pregnancies often present with symptoms during the first trimester. CASE: A previously asymptomatic woman presented with pelvic pain and vaginal bleeding at 4 months' gestation and was found to have a live, 14-week, tubal pregnancy. CONCLUSION: Second-trimester symptoms, including nausea, vomiting, pelvic or abdominal pain, and vaginal bleeding, necessitate ultrasound to determine the pregnancy location, maintaining suspicion for a tubal or abdominal pregnancy. PMID- 16363763 TI - Endocervicosis of the bladder: a case report. AB - BACKGROUND: Endocervicosis of the bladder is a rare, benign lesion characterized by mucinous endocervical epithelium within the detrusor muscle of the bladder. CASE: A 48-year-old woman presented with a history of dysuria for the past week and pelvic pain for the past 6 months. Her history was significant for 2 prior cesarean sections. Transvaginal pelvic ultrasound revealed a mass protruding into the bladder. Cystoscopy and magnetic resonance imaging of the pelvis confirmed this finding. A total abdominal hysterectomy ing. A total abdominal hysterectomy and partial cystectomy were performed. Pathology of the bladder wall revealed endocervicosis. The patient remained symptom free at 6 months. CONCLUSION: Endocervicosis of the bladder wall is a rare lesion, and the diagnosis is difficult to make both clinically and pathologically. The diagnosis should be considered in any woman presenting with pelvic pain, dysuria and frequency. Gynecologists should be aware of this condition for proper diagnostic and surgical management. PMID- 16363764 TI - Tick-borne relapsing fever during pregnancy: a case report. AB - BACKGROUND: Tick-borne relapsing fever (TBRF) is uncommon. Vertical transmission in the gravid woman may result in spontaneous abortion, preterm delivery or perinatal mortality. We report a case in a pregnant woman who experienced a Jarisch-Herxheimer reaction (J-HR). CASE: A 24-year-old woman at 23 weeks' gestation had a relapsing fever of unknown origin and nonspecific complaints. Her peripheral blood smear demonstrated spirochetes consistent with TBRF caused by Borrelia. Treatment with antibiotics precipitated J-HR. Supportive measures and appropriate antibiotic therapy resulted in a clinical cure and normal term delivery. CONCLUSION: The diagnosis of TBRF should be considered in individuals with recurrent fevers, especially those who have been in mountainous regions where Borrelia hermsii exists. Individuals receiving antimicrobials should be closely monitored for a J-HR. PMID- 16363765 TI - Vulvar Kaposi's sarcoma in a woman with AIDS: a case report. AB - BACKGROUND: Kaposi's sarcoma is the most common malignancy among HIV patients. This systemic disease has an unpredictable clinical course and commonly manifests as a cutaneous face, trunk or lower extremity lesion. CASE: A 38-year-old, multiparous woman and AIDS patient presented with left vulvar edema and pain. The patient subsequently underwent surgery for incision and drainage of a possible abscess. Operative biopsy demonstrated Kaposi's sarcoma, which was treated with palliative radiation therapy. The patient was discharged in the care of hospice. CONCLUSION: Vulvar lesions may be thefirst sign of systemic Kaposi's sarcoma in the immunocompromised woman. Most vulvar lesions warrant careful evaluation and biopsy. PMID- 16363766 TI - [Recommendations on hypertension in children: the CHI/d project]. PMID- 16363767 TI - [Renal tolerability of non-steroidal drugs (NSAIDS) in newborn infants]. PMID- 16363768 TI - A review of cases of posterior urethral valves seen at the University College Hospital, Ibadan (Nigeria). AB - A cohort of 40 male children with Posterior Urethral Valves (PUV) seen in the Paediatric Nephrology/Urology Unit of the University College Hospital, Ibadan are presented. They were reviewed with the aim of determining the clinical course of the disease in a developing country where the facilities for active intervention are not always available. Even though 50% of the patients became symptomatic in the first week of life only 22.5% presented in the whole of the neonatal period. Thirty-seven and a half per-cent (37.5%) presented in the post-neonatal infancy period and the rest beond the first year of life. The interval between the onset of symptoms and definitive therapy was up to three years in some patients. Only 2 patients had antenatal diagnosis of the PUV by ultrasonography. The major renal complications were: (1) Urinary Tract Infections in 40%; (2) Acute Renal failure- 10%; (3) Chronic Renal failure--7%; 4) Type IV Renal Tubular Acidosis--10% (5) Sustained hypertension--4.8%. The extra-renal complications were anaemia (30%), malnutrition (10%). Five of the patients died giving a case fatality rate of 12.5%, mainly from overwhelming infections and renal failure. Many of our patients had delayed presentation even though symptomatic and that may partly explain the complications and the poor outcome seen in the short term. PMID- 16363769 TI - Mycoplasma pneumonias distribution, epidemiology and prevalence in a triennial survey. AB - OBJECTIVES: To evaluate: (1) the incidence of pneumonia monthly distribution; (2) the rate of pneumonitis due to Mycoplasma Pneumoniae; (3) the suitability of choosing an empirical-based antibiotic-therapy; (4) the need of a critical revision of Mycoplasma serological data. PATIENTS AND METHODS: We studied 188 patients admitted to the Palermo University Pediatrics Department, from september 1998 to august 2001, with admission diagnosis of pneumonia. RESULTS: The highest incidence of pneumonia was in december and march as in both months 28 cases occurred in the whole period 1998-2001 (average of 9.3 cases per each month). The highest rate of pneumonias by Mycoplasma Pneumoniae was in may with a total number of 11/21 cases (52.3%, average of 3.67 cases per month) in the above mentioned three-year period. CONCLUSIONS: Incidence of Mycoplasma pneumonitis is more represented in subjects that are at school age. Our study confirms the enormous variability of the prevalence of the infection by Mycoplasma Pneumoniae and how difficult it is to make an accurate diagnosis lacking standardized, rapid, specific and comparative methods. PMID- 16363771 TI - [Visual motor and visual defects in spina bifida]. AB - Among the numerous problems that Spina Bifida (SB) patients are faced with, impairments to the visual apparatus are often considered late and are not covered extensively in the literature. At the Pediatric Department of the University of Padua, an assessment of the visual function of 59 SB patients between 5 months and 26 years of age (29 male and 30 female) has been carried out by means of an ophthalmologic protocol As far as the alteration of the exstrinsic ocular mobility is concerned, 44% (26/59) of our patients revealed a manifest squint and only 3% (two patients) suffered from a latent squint. The most frequent type is a convergent squint (80%). The assessment of visual acuity made through Optotype was good in 82% of the cases (unlike what is commonly reported in the literature) and mild in 18%. None of the patients manifested hypovision. Refraction defects were present in 59% (34/59) of the patients. Regular ophthalmologic evaluations from birth or from diagnosis allow ophthalmologic treatments that are tailored to children suffering from SB and also enable them to reach and maintain a good visual standard and to observe the subtle symptoms of endocranial hypertension sooner. An early discovery and correct treatment of visual problems improves cognitive and motive performance as well as the autonomy of SB patients. PMID- 16363770 TI - [Immunological development in asymptomatic HIV infected children]. AB - In HIV infected children, CD45+CD4+ T lymph. reconstitution has been related to efficient thymopoiesis. Because human thymus undergoes spontaneous involution at a relatively young age, institution of antiretroviral therapy early in the course of infection has been recommended. 12 HIV vertically infected children aged 4-8 years were investigated T-cell subsets for four years. 7 children were naive for therapy (group NT); 5 experienced nucleoside analogues only (group T). CD45RA+ and CD45RO+ CD4+ values were compared to predicted values of healthy children. The two groups showed similar clinical and immunological baseline characteristics (CDC class N-A). Mean VL at t0 was 4.26 log10 (SD 0.71) in gr. NT and 4.01 log10 (SD 0.57) in gr. T; median CD4 T lymph values were 27% in gr. NT and 23.5% in gr. T. Median CD45RA+ values were 62.8% in gr. NT and 71.3% in gr. T. No differences in VL, CD4+ T lymph., CD45RA+, CD45RO+ were found in between groups or within each group at each time evaluation. Median CD45RA+ values were not different from predicted values of healthy children. None of the children changed CDC class during the study period. Although the number of subjects was small, our study evidenced the possibility of a normal immunological development in HIV-1 vertically infected asymptomatic children naive for HAART during the first decade, even in the presence of significant viremia. PMID- 16363772 TI - Initial experience with laparoscopic adrenal surgery in children: is endoscopic surgery recommended and safe for the treatment of adrenocortical neoplasms? AB - Through a review of 83 cases reported in literature, including our experience of two successful right laparoscopic adrenalectomies performed in a 3-year-old girl for androgen-secreting adenoma and in a 9-year-old male for pheochromocitoma, we have anaIyzed the indications, the techniques and the results of laparoscopic or retroperitoneoscopic adrenalectomy in children. Nineteen pheochromocytomas, 18 neuroblastomas, 15 adenomas, 12 ganglioneuromas, 9 hyperplasias, 2 carcinomas, 1 teratoma and 1 adrenogenital syndrome have been treated with excellent results (follow-up 1 month to 4 years) with no evidence of recurrence. The age range is from 1 month to 16 years with an equal female/male ratio. The tumor size range from 10 mm to 80 mm with approximately a mean of 40 mm. Sixty-three laparoscopies of which 54 lateral and 9 anterior have been performed versus 14 posterior retroperitoneoscopies. Regarding right adrenalectomy, 29 were laparoscopic and only 2 retroperitoneoscopic. With regards to left adrenalectomy, 23 were laparoscopic and 9 retroperitoneoscopic. Looking at bilateral lesions, these tumors were approached laparoscopically in 6 cases alternating both sides and 1 retroperitoneoscopically (this case was already operated on the other side). The operative time ranged from 25 minutes in newborn to 320 minutes in bilateral cases with an average of 120 minutes. Conversion rate is higher in retroperitoneoscopy (30%) (4 cases out of 13), than laparoscopy (6%) (4 cases out of 63). Specifically, for right retroperitoneoscopic adrenalectomy conversion rate was 100% (2 cases out 2), for left retroperitoneoscopic adrenalectomy 22.2% (2 cases out 9), for right laparoscopic adrenalectomy 6.8% (2 cases out of 29) and for left laparoscopic adrenalectomy 8.6% (2 cases out of 23). Only in 1 case were blood transfusions requested. Hospital stays ranged from 35 hours to 17 days with an average of 4 days. In conclusion the indications of endoscopic adrenalectomy in children are not different from those of traditional surgery and the well-known advantages of laparoscopic adrenal surgery should be applied to pediatric patients. It seems that there is no age and tumor size limits for a well-trained surgical team. Lateral transperitoneal approach is the most utilized with the child positioned in 90-degree flank decubitus. Laparoscopy is undoubtly preferred for right adrenalectomy (93% of cases), while for left adrenalectomy retroperitoneoscopy has been used in 39% of the cases. Considering the conversion rate and on the basis of our experience with adults, we recommend laparoscopic adrenalectomy for both right and left adrenal lesions, but we think that the pediatric surgeon should feel free to choose the approach in which he/she is more skilled. However, the best surgical result will be achieved if the pediatric and adult surgeon collaborate with their different experiences. Lastly, we suggest the use of new technological devices such as Ultracision Harmonic Scalpel which was a critical factor in our two successful right adrenalectomies. PMID- 16363773 TI - [Hypertension in children]. PMID- 16363774 TI - Determination of optical parameters of a twisted-nematic liquid crystal by phase sensitive optical heterodyne interferometric ellipsometry. AB - What is believed to be a novel phase-sensitive optical heterodyne interferometric ellipsometer is set up to characterize a twisted-nematic liquid crystal (TN-LC) by the elliptical parameters of the output polarization state. This ellipsometer presents the advantages of both polarized optical heterodyne interferometry and optical photometry, which introduce a polarization modulation that is capable of performing with high-sensitivity on phase detection in real time. The twist angle phi and the untwisted phase retardation gamma of TN-LC are measured precisely. The experimental results verify that a TN-LC can be treated as identical to an elliptical retarder. PMID- 16363775 TI - Simple multifrequency and phase-shifting fringe-projection system based on two wavelength lateral shearing interferometry for three-dimensional profilometry. AB - We propose a simple multifrequency spatial-carrier and phase-shifting fringe projection system based on two-wavelength lateral shearing interferometry (LSI). In this system a wedge-shaped plate lateral shearing interferometer is used and, owing to the presence of tilt, a finite number of fringes parallel to the direction of the shear appears; hence a significant spatial-carrier frequency is generated at the focus position. We further enhance the spatial-carrier frequency either by changing the wavelength of the laser light or by slight defocusing. A synthetic interferogram with low spatial-carrier frequency is obtained by use of laser light of two wavelengths simultaneously in the lateral shear interferometer. We obtain the phase-shifted fringe patterns from the same setup by simply moving the wedge plate in an in-plane parallel direction, using a linear translator. The fringe projection system was tested for measurement of the three-dimensional shape of a discontinuous object. The present system has many advantages; e.g., it is a common-path interferometry and hence is insensitive to external vibrations, is compact in size, and is relatively inexpensive. PMID- 16363776 TI - Mode propagation in optical nanowaveguides with dielectric cores and surrounding metal layers. AB - The mode spectrum in an optical nanowaveguide consisting of a dielectric-core layer surrounded by two identical metal layers is investigated. A simple model based on mode matching to predict the properties of mode propagation in such optical nanowaveguides is proposed. It is shown that quasi-TM00 and quasi-TM10 modes supported by an optical microstrip line do not have a cutoff frequency, regardless of the size of the metal strips, the thickness of the dielectric slab, and the cross-sectional shape. The transverse size of the TM00 mode supported by a nanosized microstrip line was found to be approximately equal to the transverse dimension of the microstrip line. In closed rectangular and elliptical nanowaveguides, i.e., in which all dielectric surfaces are covered with metal films, the cross-sectional shape of the waveguide should be stretched along one side to produce propagation conditions for the fundamental mode. PMID- 16363777 TI - Generalized matrix method for calculation of internal light energy flux in mixed coherent and incoherent multilayers. AB - A generalized matrix method to treat multilayer systems with mixed coherent and incoherent optical behavior is presented. The method is based on the calculation of the light energy flux inside the multilayer, whose internal light absorption is straightforwardly derived. The Poynting vector is used to derive the light energy flux in the case of a layer with coherent behavior. Multilayer structures with any distribution of layers with coherent or incoherent behavior can be treated, including the case of oblique incidence. Use of the light energy flux instead of the more commonly used light intensity permits the calculation of light absorption with a better accuracy and a much shorter computation time. PMID- 16363778 TI - Compact double-grating coupler between vertically stacked silicon-on-insulator waveguides. AB - We analyze a compact double-grating coupler that provides coupling through radiation modes between two vertically stacked silicon-on-insulator waveguides. The grating is sufficiently strong to be considered a one-dimensional photonic bandgap structure that facilitates a short coupling length. Simulations suggest that a 29% coupling efficiency is achievable in coupling light from one waveguide to another with 12.9 microm long binary gratings. We found that the coupling efficiency is enhanced by Fabry-Perot resonance between two gratings. The coupling efficiency can be increased by use of a blazed grating. We use the eigenmode expansion method to design and optimize the binary grating coupler, and the results are verified by use of the finite-difference time-domain method. PMID- 16363779 TI - Effects of birefringence on Fizeau interferometry that uses a polarization phase shifting technique. AB - Interferometers that use different states of polarization for the reference and the test beams can modulate the relative phase shift by using polarization optics in the imaging system. Thus the interferometer can capture simultaneous images that have a fixed phase shift, which can be used for phase-shifting interferometry. As all measurements are made simultaneously, the interferometer is not sensitive to vibration. Fizeau interferometers of this type have an advantage compared with Twyman-Green-type systems because they are common-path interferometers. However, a polarization Fizeau interferometer is not strictly common path when both wavefronts are transmitted by an optic that suffers from birefringence. The two polarized beams see different phases owing to birefringence; as a result, an error can be introduced in the measurement. We study the effect of birefringence on measurement accuracy when different polarization techniques are used in Fizeau interferometers. We demonstrate that measurement error is reduced dramatically and can be eliminated if the reference and test beams are circularly polarized rather than linearly polarized. PMID- 16363780 TI - Simple algorithm for the measurement of absorption coefficients of a two-layered medium by spatially resolved and time-resolved reflectance. AB - An inversion procedure for the recovery of absorption coefficients of a two layered semi-infinite diffusive medium by use of time-resolved reflectance measured at two different source-detector distances is proposed. The inversion procedure is based on the property of the photon diffusion equation; i.e., the solution of the diffusion equation for the time-resolved reflectance measured at a longer source--detector distance coincides with that measured at a shorter one by a proper temporal, spatial, and intensity transformation. This inversion procedure, used together with the results of one set of Monte Carlo simulations, is validated as working well when the values of the scattering coefficients of the two layers and the thickness of the first layer are within a range of interest in tissue optics. PMID- 16363781 TI - Time-resolved Hadamard fluorescence imaging. AB - We present a new concept for fluorescence lifetime imaging (FLIM) based on time resolved Hadamard imaging (HI). HI allows image acquisition by use of one single point detector without requiring a moving scanning stage. Moreover, it reduces the influence of detector noise compared with raster scanning. By use of Monte Carlo simulations it could be confirmed that Hadamard transformation may decrease the error in lifetime estimation and in general in fluorescence parameter estimation when the signal-to-noise ratio is low and detector dark noise is high. This concept may find applications whenever the performance of FLIM or similar methods is limited by high dark-count rates and when the use of a single-point detector is preferable. PMID- 16363782 TI - Cascaded holographic polymer reflection grating filters for optical-code-division multiple-access applications. AB - We evaluate the use of edge-illuminated holographic Bragg filters formed in phenanthrenequinone-doped poly(methyl methacrylate) for optical-code-division multiple-access (OCDMA) coding and decoding applications. Experimental cascaded Bragg filters are formed to select two different wavelengths with a fixed distance between the gratings and are directly coupled to a fiber-measurement system. The configuration and tolerances of the cascaded gratings are shown to be practical for time-wavelength OCDMA applications. PMID- 16363783 TI - Atmospheric wavefront phase recovery by use of specialized hardware: graphical processing units and field-programmable gate arrays. AB - To achieve the wavefront phase-recovery stage of an adaptive-optics loop computed in real time for 32 x 32 or a greater number of subpupils in a Shack-Hartmann sensor, we present here, for what is to our knowledge the first time, preliminary results that we obtained by using innovative techniques: graphical processing units (GPUs) and field-programmable gate arrays (FPGAs). We describe the stream computing paradigm of the GPU and adapt a zonal algorithm to take advantage of the parallel computational power of the GPU. We also present preliminary results we obtained by use of FPGAs on the same algorithm. GPUs have proved to be a promising technique, but FPGAs are already a feasible solution to adaptive-optics real-time requirements, even for a large number of subpupils. PMID- 16363784 TI - High-order cumulant-based blind deconvolution of Raman spectra. AB - The spectra recorded by a dispersion spectrophotometer are usually distorted by the response function of the instrument. To improve the resolving power, double or triple cascade spectrophotometers with narrow slits have been employed, but the total flux of the radiation available decreases accordingly, resulting in a low signal-to-noise ratio and a longer measuring time. The actual spectra can be restored approximately by mathematically removing the effects of the measuring instruments. Based on the Shalvi-Weinstein criterion, a (6, 2)-order normalized cumulant-based blind deconvolution algorithm for Raman spectral data is proposed. The actual spectral data and the unit-impulse response of the measuring instruments can be estimated simultaneously. By conducting experiments on real Raman spectra of some organic compounds, it is shown that this algorithm has a robust performance and fast convergence behavior and can improve the resolving power and correct the relative intensity distortion considerably. PMID- 16363785 TI - Parameterized code SHARM-3D for radiative transfer over inhomogeneous surfaces. AB - The code SHARM-3D, developed for fast and accurate simulations of the monochromatic radiance at the top of the atmosphere over spatially variable surfaces with Lambertian or anisotropic reflectance, is described. The atmosphere is assumed to be laterally uniform across the image and to consist of two layers with aerosols contained in the bottom layer. The SHARM-3D code performs simultaneous calculations for all specified incidence-view geometries and multiple wavelengths in one run. The numerical efficiency of the current version of code is close to its potential limit and is achieved by means of two innovations. The first is the development of a comprehensive precomputed lookup table of the three-dimensional atmospheric optical transfer function for various atmospheric conditions. The second is the use of a linear kernel model of the land surface bidirectional reflectance factor (BRF) in our algorithm that has led to a fully parameterized solution in terms of the surface BRF parameters. The code is also able to model inland lakes and rivers. The water pixels are described with the Nakajima-Tanaka BRF model of wind-roughened water surface with a Lambertian offset, which is designed to model approximately the reflectance of suspended matter and of a shallow lake or river bottom. PMID- 16363786 TI - Picosecond soliton transmission by use of concatenated gain-distributed nonlinear amplifying fiber loop mirrors. AB - Stable picosecond soliton transmission is demonstrated numerically by use of concatenated gain-distributed nonlinear amplifying fiber loop mirrors (NALMs). We show that, as compared with previous soliton transmission schemes that use conventional NALMs or nonlinear optical loop mirror and amplifier combinations, the present scheme permits a significant increase of loop-mirror (amplifier) spacing. The broad switching window of the present device and the high-quality pulses switched from it provide a reasonable stability range for soliton transmission. We also show that a soliton self-frequency shift can be suppressed by the gain-dispersion effect in the amplifying fiber loop and that soliton soliton interactions can be partially reduced by using lowly dispersive transmission fibers. PMID- 16363787 TI - Synthetic-aperture imaging laser radar: laboratory demonstration and signal processing. AB - The spatial resolution of a conventional imaging laser radar system is constrained by the diffraction limit of the telescope's aperture. We investigate a technique known as synthetic-aperture imaging laser radar (SAIL), which employs aperture synthesis with coherent laser radar to overcome the diffraction limit and achieve fine-resolution, long-range, two-dimensional imaging with modest aperture diameters. We detail our laboratory-scale SAIL testbed, digital signal processing techniques, and image results. In particular, we report what we believe to be the first optical synthetic-aperture image of a fixed, diffusely scattering target with a moving aperture. A number of fine-resolution, well focused SAIL images are shown, including both retroreflecting and diffuse scattering targets, with a comparison of resolution between real-aperture imaging and synthetic-aperture imaging. A general digital signal-processing solution to the laser waveform instability problem is described and demonstrated, involving both new algorithms and hardware elements. These algorithms are primarily data driven, without a priori knowledge of waveform and sensor position, representing a crucial step in developing a robust imaging system. PMID- 16363788 TI - Suppression of nonlinear frequency sweep in an optical frequency-domain reflectometer by use of Hilbert transformation. AB - A compensation technique for reducing the effect of nonlinear optical frequency swept in an optical frequency-domain reflectometer (OFDR) is proposed. The instantaneous sweep optical frequency of an OFDR laser source is directly obtained by analysis of the interference signal from an auxiliary interferometer with a Hilbert transformation. Beating OFDR data from a main interferometer are regenerated with respect to the measured instantaneous optical frequency. We show that this technique dramatically improves the spatial resolution of a conventional OFDR and can be applied to an optical frequency-domain medical imaging system to eliminate the problem of a nonlinear frequency sweep effect. PMID- 16363789 TI - Stable two-wavelength lasers by use of a double alpha-type fiber cavity with fiber grating mirrors. AB - For high-frequency (20-200 GHz) modulated light sources, we developed and investigated two-wavelength lasers using the double alpha-type fiber cavities with fiber grating mirrors. For variations of polarization states and pump powers, parallel alpha-type coupled cavity lasers were found to be more stable than serial lasers. PMID- 16363790 TI - Acceptable composition-ratio variations of a mixed crystal for nonlinear laser device applications. AB - A theory is developed to predict some crucial parameters that optimize the performance of mixed nonlinear crystals in nonlinear devices. These include acceptable variations of the composition ratio of the parent crystals and the optimal as well as acceptable interaction lengths for any interaction. The theory is successfully applied to make necessary predictions for the newly developed LiIn(Se(x)S(1-x))2 crystal for second-harmonic and optical parametric generation. PMID- 16363791 TI - Monolithic wavelength converter for ultraviolet light by use of a Gd(x)Y(1 x)Ca4O(BO3)3 crystal. AB - Gd(x)Y(1-x)Ca4O(BO3)3 (GdYCOB) is a promising nonlinear optical crystal that shows high effective nonlinearity d(eff), noncritical phase matching, and high chemical stability. We report on the fabrication and characteristics of a monolithic wavelength converter, which generates ultraviolet light by the incidence of a 1.064 microm near-infrared laser. The converter consists of GdYCOB for third-harmonic generation, KTiOPO4 (KTP) for second-harmonic generation, and a wave plate. GdYCOB has the advantage of an extremely wide angular acceptance bandwidth, whereas KTP exhibits a high effective nonlinear coefficient and a broad temperature bandwidth. Consequently the combination of these crystals results in highly efficient and stable ultraviolet conversion for constructing a compact and robust ultraviolet laser. PMID- 16363792 TI - The heme of cystathionine beta-synthase likely undergoes a thermally induced redox-mediated ligand switch. AB - Cystathionine beta-synthase (CBS) is a pyridoxal-5'-dependent enzyme that catalyzes the condensation of homocysteine and serine to form cystathionine. Human CBS is unique in that heme is also required for maximal activity, although the function of heme in this enzyme is presently unclear. The study presented herein reveals that the heme of human CBS undergoes a coordination change upon reduction at elevated temperatures. We have termed this new species "CBS424" and demonstrate that its formation is likely irreversible when pH 9 Fe(III) CBS is reduced at moderately elevated temperatures (approximately 40 degrees C and higher) or when pH 9 Fe(II) CBS is heated to similar temperatures. Spectroscopic techniques, including resonance Raman, electronic absorption, and variable temperature/variable field magnetic circular dichroism spectroscopy, provide strong evidence that CBS424 is coordinated by two neutral donor ligands. It appears likely that the native cysteine(thiolate) heme ligand is displaced by an endogenous neutral donor upon conversion to CBS424. This behavior is consistent with other six-coordinate, cysteine(thiolate)-ligated heme centers, which seek to avoid this coordination structure in the Fe(II) state. Functional assays show that CBS424 is inactive and suggest that the ligand switch is responsible for eliminating enzyme activity. When this investigation is taken together with other functional studies of CBS, it provides strong evidence that coordination of Cys52 to the heme iron is crucial for full activity in this enzyme. We hypothesize that cysteine displacement may serve as a mechanism for CBS inactivation and that second-sphere interactions of the Cys52 thiolate with surrounding residues are responsible for communicating the heme ligand displacement to the CBS active site. PMID- 16363793 TI - Structure of homeodomain-leucine zipper/DNA complexes studied using hydroxyl radical cleavage of DNA and methylation interference. AB - Homeodomain-leucine zipper (HD-Zip) proteins, unlike most homeodomain proteins, bind a pseudopalindromic DNA sequence as dimers. We have investigated the structure of the DNA complexes formed by two HD-Zip proteins with different nucleotide preferences at the central position of the binding site using footprinting and interference methods. The results indicate that the respective complexes are not symmetric, with the strand bearing a central purine (top strand) showing higher protection around the central region and the bottom strand protected toward the 3' end. Binding to a sequence with a nonpreferred central base pair produces a decrease in protection in either the top or the bottom strand, depending upon the protein. Modeling studies derived from the complex formed by the monomeric Antennapedia homeodomain with DNA indicate that in the HD Zip/DNA complex the recognition helix of one of the monomers is displaced within the major groove respective to the other one. This monomer seems to lose contacts with a part of the recognition sequence upon binding to the nonpreferred site. The results show that the structure of the complex formed by HD-Zip proteins with DNA is dependent upon both protein intrinsic characteristics and the nucleotides present at the central position of the recognition sequence. PMID- 16363794 TI - Time-resolved single tryptophan fluorescence in photoactive yellow protein monitors changes in the chromophore structure during the photocycle via energy transfer. AB - We show from time-resolved fluorescence intensity and depolarization experiments that the fluorescence of the unique tryptophan W119 of PYP is quenched by energy transfer to the 4-hydroxycinnamoyl chromophore. Whereas the intensity decay has a time constant of 0.18 ns in P, the decay in the absence of the cofactor (apo-PYP) has a single exponential lifetime of 4.8 ns. This difference in lifetime with and without acceptor can be explained quantitatively on the basis of energy transfer and the high-resolution X-ray structure of P, which allows an accurate calculation of the kappa2 factor. Fluorescence depolarization experiments with donor and acceptor indicate that both are immobilized so that kappa2 is constant on the fluorescence time scale. Using background illumination from an LED emitting at 470 nm, we measured the time-resolved fluorescence in a photostationary mixture of P and the intermediates I2 and I2'. The composition of the photostationary mixture depends on pH and changes from mainly I2 at low pH to predominantly I2' at high pH. The I2/I2' equilibrium is pH-dependent with a pKa of approximately 6.3. In I2 the lifetime increases to approximately 0.82 ns. This is not due to a change in distance or to the increase in spectral overlap but is primarily a consequence of a large decrease in kappa2. Kappa2 was calculated from the available X-ray structures and decreases from approximately 2.7 in P to 0.27 in I2. This change in kappa2 is caused by the isomerization of the acceptor, which leads to a reorientation of its transition dipole moment. We have here a rare case of the kappa2 factor dominating the change in energy transfer. The fluorescence decay in the light is pH-dependent. From an SVD analysis of the light/dark difference intensity decay at a number of pH values, we identify three species with associated lifetimes: P (0.18 ns), I2 (0.82 ns), and X (0.04 ns). On the basis of the pH dependence of the amplitudes associated with I2 and X, with a pKa of approximately 6.3, we assign the third species to the signaling state I2'. The absorption spectra of the 0.82 and 0.04 ns species were calculated from the pH dependence of their fluorescence amplitudes and of the photostationary light/dark difference absorption spectra. The lambda(max) values of these spectra (372 and 352 nm) identify the 0.82 and 0.04 ns components with I2 and I2', respectively, and validate the fluorescence data analysis. The mutant E46Q allows a further test of the energy transfer explanation, since lowering the pH in the dark leads to a bleached state with an increased spectral overlap but without the isomerization-induced decrease in kappa2. The measured lifetime of 0.04 ns is in excellent agreement with predictions based on energy transfer and the X-ray structure. PMID- 16363795 TI - Effects of charge-to-alanine substitutions on the stability of ribosomal protein L30e from Thermococcus celer. AB - The ability to rationally engineer a protein with altered stability depends upon the detailed understanding of the role of noncovalent interactions in defining thermodynamic properties of proteins. In this paper, we used T. celer L30e as a model to address the question of the role of charge-charge interactions in defining the stability of this protein. A total of 26 single-site charge-to alanine variants of this protein were generated, and the stability of these proteins was determined using thermal- and denaturant-induced unfolding. It was found that, although L30e is isolated from a thermophilic organism and is highly thermostable, some of the substitutions lead to a further increase in the transition temperature. Analysis of the effects of high ionic strength on the stabilities of L30e variants shows that the long-range charge-charge interactions are as important as the short-range (salt bridge) interactions. The changes in stabilities of the T. celer L30e protein variants were compared with the changes in the energy of charge-charge interactions calculated using different computational models. It was found that there is a good qualitative agreement between experimental and calculated data: for 70-80% (19-21 of 26, confidence p < 0.003) of the variants, computational models predict correctly the sign of the stability changes. In particular, computational models identify correctly those charged amino acid residue substitutions of which led to enhancement in thermostability. Thus, optimization of the charge-charge interactions might be a useful approach for the rational increase in protein stability. PMID- 16363796 TI - Distinct transport selectivity of two structural subclasses of the nodulin-like intrinsic protein family of plant aquaglyceroporin channels. AB - Major intrinsic proteins (MIPs) are a diverse class of integral membrane proteins that facilitate the transport of water and some small solutes across cellular membranes. X-ray structures of MIPs indicate that a tetrad of residues (the ar/R region) form a narrow pore constriction that constitutes the selectivity filter. In comparison with mammalian and microbial species, plants have a greater number and diversity of MIPs with greater than 30 genes encoding four phylogenetic subfamilies with eight different classes of ar/R sequences. The nodulin 26-like intrinsic protein (NIP) subfamily in Arabidopsis can be subdivided into two ar/R subgroups: the NIP subgroup I, which resembles the archetype of the family, soybean nodulin 26, and the NIP subgroup II, which is represented by the Arabidopsis protein AtNIP6;1. These two NIPs differ principally by the substitution of a conserved alanine (NIP subgroup II) for a conserved tryptophan (NIP subgroup I) in the helix 2 position (H2) of the ar/R filter. A comparison of the water and solute tranport properties of the two proteins was performed by expression in Xenopus laevis oocytes. Nodulin 26 is an aquaglyceroporin with a modest osmotic water permeability (P(f)) and the ability to transport uncharged solutes such as glycerol and formamide. In constrast, AtNIP6;1 showed no measurable water permeability but transported glycerol, formamide, as well as larger solutes that were impermeable to nodulin 26. By site-directed mutagenesis, we show that the H2 position is the crucial determinant that confers these transport behaviors. A comparison of the NIPs and tonoplast-intrinsic proteins (TIP) shows that the H2 residue can predict the transport profile for water and glycerol with histidine found in TIP-like aquaporins, tryptophan found in aquaglyceroporins (NIP I), and alanine found in water-impermeable glyceroporins (AtNIP6;1). PMID- 16363797 TI - Conformation coupled enzyme catalysis: single-molecule and transient kinetics investigation of dihydrofolate reductase. AB - Ensemble kinetics and single-molecule fluorescence microscopy were used to study conformational transitions associated with enzyme catalysis by dihydrofolate reductase (DHFR). The active site loop of DHFR was labeled with a fluorescence quencher, QSY35, at amino acid position 17, and the fluorescent probe, Alexa555, at amino acid 37, by introducing cysteines at these sites with site-specific mutagenesis. The distance between the probes was such that approximately 50% fluorescence resonance energy transfer (FRET) occurred. The double-labeled enzyme retained essentially full catalytic activity, and stopped-flow studies of both the forward and reverse reactions revealed that the distance between probes increased prior to hydride transfer. A fluctuation in fluorescence intensity of single molecules of DHFR was observed in an equilibrium mixture of substrates but not in their absence. Ensemble rate constants were derived from the distributions of lifetimes observed and attributed to a reversible conformational change. Studies were carried out with both NADPH and NADPD as substrates, with no measurable isotope effect. Similar studies with a G121V mutant DHFR resulted in smaller rate constants. This mutant DHFR has reduced catalytic activity, so that the collective data for the conformational change suggest that the conformational change being observed is associated with catalysis and probably represents a conformational change prior to hydride transfer. If the change in fluorescence is attributed to a change in FRET, the distance change associated with the conformational change is approximately 1-2 A. These results are correlated with other measurements related to conformation coupled catalysis. PMID- 16363798 TI - Identification of amino acid residues contributing to the mechanism of cooperativity in Escherichia coli D-3-phosphoglycerate dehydrogenase. AB - L-Serine inhibits the catalytic activity of Escherichia coli D-3-phosphoglycerate dehydrogenase (PGDH) by binding to its regulatory domain. This domain is a member of the ACT domain family of regulatory domains that are modulated by small molecules. A comparison of the phi and psi torsional angle differences between the crystal structures of PGDH solved in the presence and in the absence of L serine demonstrated a clustering of significant angle deviations in the regulatory domain. A similar clustering was not observed in either of the other two structural domains of PGDH. In addition, significant differences were also observed at the active site and in the Trp-139 loop. To determine if these residues were functionally significant and not just due to other factors such as crystal packing, mutagenic analysis of these residues was performed. Not unexpectedly, this analysis showed that residues that affected the kcat/Km were grouped around the active site and those that affected the serine sensitivity were grouped in the regulatory domain. However, more significantly, residues that affected the cooperativity of inhibition of activity were identified at both locations. These latter residues represent structural elements that participate in both the initial and the ultimate events of the transfer of cooperative behavior from the regulatory domain to the active site. As such, their identification will assist in the elucidation of the pathway of cooperative interaction in this enzyme as well as in the elucidation of the regulatory mechanism of the ACT domain in general. PMID- 16363799 TI - Substrate-assisted movement of the catalytic Lys 215 during domain closure: site directed mutagenesis studies of human 3-phosphoglycerate kinase. AB - 3-Phosphoglycerate kinase (PGK) is a two-domain hinge-bending enzyme. It is still unclear how the geometry of the active site is formed during domain closure and how the catalytic residues are brought into the optimal position for the reaction. Comparison of the three-dimensional structures in various open and closed conformations suggests a large (10 A) movement of Lys 215 during domain closure. This change would be required for direct participation of this side chain in both the catalyzed phospho transfer and the special anion-caused activation. To test the multiple roles of Lys 215, two mutants (K215A and K215R) were constructed from human PGK and characterized in enzyme kinetic and substrate binding studies. For comparison, mutants (R38A and R38K) of the known essential residue, Arg 38, were also produced. Drastic decreases (1500- and 500-fold, respectively), as in the case of R38A, were observed in the kcat values of mutants K215A and K215R, approving the essential catalytic role of Lys 215. In contrast, the R38K mutation caused an only 1.5-fold decrease in activity. This emphasizes the importance of a very precise positioning of Lys 215 in the active site, in addition to its positive charge. The side chain of Lys 215 is also responsible for the substrate and anion-dependent activation, since these properties are abolished upon mutation. Among the kinetic constants mainly the Km values of MgATP and 1,3-BPG are increased (approximately 20- and approximately 8 fold, respectively) in the case of the neutral K215A mutant, evidence of the interaction of Lys 215 with the transferring phospho group in the functioning complex. Weakening of MgATP binding (a moderate increase in Kd), but not of MgADP binding, upon mutation indicates an initial weak interaction of Lys 215 with the gamma-phosphate already in the nonfunctioning open conformation. Thus, during domain closure, Lys 215 possibly moves together with the transferring phosphate; meanwhile, this group is being positioned properly for catalysis. PMID- 16363800 TI - Ionization of zwitterionic amine substrates bound to monomeric sarcosine oxidase. AB - Monomeric sarcosine oxidase (MSOX) binds the L-proline zwitterion (pKa = 10.6). The reactive substrate anion is generated by ionization of the ES complex (pKa = 8.0). Tyr317 was mutated to Phe to determine whether this step might involve proton transfer to an active site base. The mutation does not eliminate the ionizable group in the ES complex (pKa = 8.9) but does cause a 20-fold decrease in the maximum rate of the reductive half-reaction. Kinetically determined Kd values for the ES complex formed with L-proline agree with results obtained in spectral titrations with the wild-type or mutant enzyme. Unlike the wild-type enzyme, Kd values with the mutant enzyme are pH-dependent, suggesting that the mutation has perturbed the pKa of a group that affects the Kd. As compared with the wild-type enzyme, an increase in charge transfer band energy is observed for mutant enzyme complexes with substrate analogues while a 10-fold decrease in the charge transfer band extinction coefficient is found for the complex with the L proline anion. The results eliminate Tyr317 as a possible acceptor of the proton released upon substrate ionization. Since previous studies rule out the only other nearby base, we conclude that L-proline is the ionizable group in the ES complex and that amino acids are activated for oxidation upon binding to MSOX by stabilization of the reactive substrate anion. Tyr317 may play a role in substrate activation and optimizing binding, as judged by the effects of its mutation on the observed pKa, reaction rates, and charge transfer bands. PMID- 16363801 TI - Mode of binding of methyl acceptor substrates to the adrenaline-synthesizing enzyme phenylethanolamine N-methyltransferase: implications for catalysis. AB - Here we report three crystal structure complexes of human phenylethanolamine N methyltransferase (PNMT), one bound with a substrate that incorporates a flexible ethanolamine side chain (p-octopamine), a second bound with a semirigid analogue substrate [cis-(1R,2S)-2-amino-1-tetralol, cis-(1R,2S)-AT], and a third with trans-(1S,2S)-2-amino-1-tetralol [trans-(1S,2S)-AT] that acts as an inhibitor of PNMT rather than a substrate. A water-mediated interaction between the critical beta-hydroxyl of the flexible ethanolamine group of p-octopamine and an acidic residue, Asp267, is likely to play a key role in positioning the side chain correctly for methylation to occur at the amine. A second interaction with Glu219 may play a lesser role. Catalysis likely occurs via deprotonation of the amine through the action of Glu185; mutation of this residue significantly reduced the kcat without affecting the Km. The mode of binding of cis-(1R,2S)-AT supports the notion that this substrate is a conformationally restrained analogue of flexible PNMT substrates, in that it forms interactions with the enzyme similar to those observed for p-octopamine. By contrast, trans-(1S,2S)-AT, an inhibitor rather than a substrate, binds in an orientation that is flipped by 180 degrees compared with cis-(1R,2S)-AT. A consequence of this flipped binding mode is that the interactions between the hydroxyl and Asp267 and Glu219 are lost. However, the amines of inhibitor trans-(1S,2S)-AT and substrate cis-(1R,2S)-AT are both within methyl transfer distance of the cofactor. These results suggest that PNMT catalyzes transfer of methyl to ligand amines only when "anchor" interactions, such as those identified for the beta-hydroxyls of p-octopamine and cis-AT, are present. PMID- 16363802 TI - Synergistic effects on escape of a ligand from the closed tryptophan synthase bienzyme complex. AB - Substrate channeling in the tryptophan synthase bienzyme complex is regulated by allosteric signals between the alpha- and beta-active sites acting over a distance of 25 A. At the alpha-site, indole is cleaved from 3-indole-D-glycerol 3'-phosphate (IGP) and is channeled to the beta-site via a tunnel. Harris and Dunn [Harris, R. M., and Dunn, M. F. (2002) Biochemistry 41, 9982-9990] showed that when the novel amino acid, dihydroiso-L-tryptophan (DIT), reacts with the beta-site, the alpha-aminoacrylate Schiff base, E(A-A), is formed and the enzyme releases indoline. The indoline produced exits the enzyme via the tunnel out the open alpha-site. When the alpha-site ligand (ASL) alpha-D,L-glycerol 3-phosphate (GP) binds and closes the alpha-site, indoline generated in the DIT reaction is trapped for a short period of time as the quinonoid intermediate in rapid equilibrium with bound indoline and the E(A-A) intermediate before leaking out of the closed enzyme. In this work, we use the DIT reaction and a new, high affinity, ASL, N-(4-trifluoromethoxybenzenesulfonyl)-2-amino-1-ethyl phosphate (F9), to explore the mechanism of ligand leakage from the closed enzyme. It was found that F9 binding to the alpha-site is significantly more effective than GP in trapping indoline in the DIT reaction; however, leakage of indoline from the enzyme into solution still occurs. It was also found that a combination of benzimidazole (BZI) and GP provided even more effective trapping than F9. The new experiments with F9 and the combination of BZI and GP provide evidence that the coincident binding of GP and BZI at the alpha-site exhibits a strong synergistic effect that greatly slows the leakage of indoline in the DIT reaction and enhances the trapping effect. This synergism functions to tightly close the alpha site and sends an allosteric signal that stabilizes the closed structure of the beta-site. These studies also support a mechanism for the escape of indoline through the alpha-site that is limited by ASL dissociation. PMID- 16363804 TI - Peptergents: peptide detergents that improve stability and functionality of a membrane protein, glycerol-3-phosphate dehydrogenase. AB - Toward enhancing in vitro membrane protein studies, we have utilized small self assembling peptides with detergent properties ("peptergents") to extract and stabilize the integral membrane flavoenzyme, glycerol-3-phosphate dehydrogenase (GlpD), and the soluble redox flavoenzyme, NADH peroxidase (Npx). GlpD is a six transmembrane spanning redox enzyme that catalyzes the oxidation of glycerol-3 phosphate to dihydroxyacetone phosphate. Although detergents such as n-octyl-beta D-glucpyranoside can efficiently solubilize the enzyme, GlpD is inactivated within days once reconstituted into detergent micelles. In contrast, peptergents can efficiently extract and solubilize GlpD from native Escherichia coli membrane and maintain its enzymatic activity up to 10 times longer than in traditional detergents. Intriguingly, peptergents also extended the activity of a soluble flavoenzyme, Npx, when used as an additive. Npx is a flavoenzyme that catalyzes the two-electron reduction of hydrogen peroxide to water using a cysteine sulfenic acid as a secondary redox center. The lability of the peroxidase results from oxidation of the sulfenic acid to the sulfinic or sulfonic acid forms. Oxidation of the sulfenic acid, the secondary redox center, results in inactivation, and this reaction proceeds in vitro even in the presence of reducing agents. Although the exact mechanism by which peptergents influence solution stability of Npx remains to be determined, the positive effects may be due to antioxidant properties of the peptides. Peptide-based detergents can be beneficial for many applications and may be particularly useful for structural and functional studies of membrane proteins due to their propensity to enhance the formation of ordered supramolecular assemblies. PMID- 16363803 TI - Use of urea and glycine betaine to quantify coupled folding and probe the burial of DNA phosphates in lac repressor-lac operator binding. AB - Thermodynamic analysis of urea-biopolymer interactions and effects of urea on folding of proteins and alpha-helical peptides shows that urea interacts primarily with polar amide surface. Urea is therefore predicted to be a quantitative probe of coupled folding, remodeling, and other large-scale changes in the amount of water-accessible polar amide surface in protein processes. A parallel analysis indicates that glycine betaine [N,N,N-trimethylglycine (GB)] can be used to detect burial or exposure of anionic (carboxylate, phosphate) biopolymer surface. To test these predictions, we have investigated the effects of these solutes (0-3 m) on the formation of 1:1 complexes between lac repressor (LacI) and its symmetric operator site (SymL) at a constant KCl molality. Urea reduces the binding constant K(TO) [initial slope dlnK(TO)/dm(urea) = -1.7 +/- 0.2], and GB increases K(TO) [initial slope dlnK(TO)/dm(GB) = 2.1 +/- 0.2]. For both solutes, this derivative decreases with an increase in solute concentration. Analysis of these initial slopes predicts that (1.5 +/- 0.3) x 10(3) A2 of polar amide surface and (4.5 +/- 1.0) x 10(2) A2 of anionic surface are buried in the association process. Analysis of published structural data, together with modeling of unfolded regions of free LacI as extended chains, indicates that 1.5 x 10(3) A2 of polar amide surface and 6.3 x 10(2) A2 of anionic surface are buried in complexation. Quantitative agreement between structural and thermodynamic results is obtained for amide surface (urea); for anionic surface (GB), the experimental value is approximately 70% of the structural value. For LacI-SymL binding, two-thirds of the structurally predicted change in amide surface (1.0 x 10(3) A2) occurs outside the protein-DNA interface in protein protein interfaces formed by folding of the hinge helices and interactions of the DNA binding domain (DBD) with the core of the repressor. Since urea interacts principally with amide surface, it is particularly well-suited to detect and quantify the extent of coupled folding and other large-scale remodeling events in the steps of protein-nucleic acid interactions and other protein associations. PMID- 16363805 TI - A cluster of basic amino acid residues in the gamma370-381 sequence of fibrinogen comprises a binding site for platelet integrin alpha(IIb)beta3 (glycoprotein IIb/IIIa). AB - Adhesive interactions of platelet integrin alpha(IIb)beta3 with fibrinogen and fibrin are central events in hemostasis and thrombosis. However, the mechanisms by which alpha(IIb)beta3 binds these ligands remain incompletely understood. We have recently demonstrated that alpha(IIb)beta3 binds the gamma365-383 sequence in the gammaC-domain of fibrin(ogen). This sequence contains neither the AGDV nor the RGD recognition motifs, known to bind alpha(IIb)beta3, suggesting the different specificity of the integrin. Here, using peptide arrays, mutant fibrinogens, and recombinant mutant gammaC-domains, we have examined the mechanism whereby alpha(IIb)beta3 binds gamma365-383. The alpha(IIb)beta3-binding activity was localized within gamma370-381, with two short sequences, gamma370ATWKTR375 and gamma376WYSMKK381, being able to independently bind the integrin. Furthermore, recognition of alpha(IIb)beta3 by gamma370-381 depended on four basic residues, Lys373, Arg375, Lys380, and Lys381. Simultaneous replacement of these amino acids and deletion of the gamma408AGDV411 sequence in the recombinant gammaC-domain resulted in the loss of alpha(IIb)beta3-mediated platelet adhesion. Confirming the critical roles of the identified residues, abnormal fibrinogen Kaiserslautern, in which gammaLys380 is replaced by Asn, demonstrated delayed clot retraction and impaired alpha(IIb)beta3 binding. Also, a mutant recombinant fibrinogen modeled after the naturally occurring variant Osaka V (gammaArg375 --> Gly) showed delayed clot retraction and reduced binding to purified alpha(IIb)beta3. These results identify the gamma370-381 sequence of fibrin(ogen) as the binding site for alpha(IIb)beta3 involved in platelet adhesion and clot retraction and define the new recognition specificity of this integrin. PMID- 16363806 TI - Proton motive force-dependent Hoechst 33342 transport by the ABC transporter LmrA of Lactococcus lactis. AB - The fluorescent compound Hoechst 33342 is a substrate for many multidrug resistance (MDR) transporters and is widely used to characterize their transport activity. We have constructed mutants of the adenosine triphosphate (ATP) binding cassette (ABC)-type MDR transporter LmrA of Lactococcus lactis that are defective in ATP hydrolysis. These mutants and wild-type LmrA exhibited an atypical behavior in the Hoechst 33342 transport assay. In membrane vesicles, Hoechst 33342 transport was shown to be independent of the ATPase activity of LmrA, and it was not inhibited by orthovanadate but sensitive to uncouplers that collapse the proton gradient and to N,N'-dicyclohexylcarbodiimide, an inhibitor of the F0F1-ATPase. In contrast, transport of Hoechst 33342 by the homologous, heterodimeric MDR transporter LmrCD showed a normal ATP dependence and was insensitive to uncouplers of the proton gradient. With intact cells, expression of LmrA resulted in an increased rate of Hoechst 33342 influx while LmrCD caused a decrease in the rate of Hoechst 33342 influx. Cellular toxicity assays using a triple knockout strain, i.e., L. lactis delta lmrA delta lmrCD, demonstrate that expression of LmrCD protects cells against the growth inhibitory effects of Hoechst 33342, while in the presence of LmrA, cells are more susceptible to Hoechst 33342. Our data demonstrate that the LmrA-mediated Hoechst 33342 transport in membrane vesicles is influenced by the transmembrane pH gradient due to a pH-dependent partitioning of Hoechst 33342 into the membrane. PMID- 16363807 TI - The PsbU subunit of photosystem II stabilizes energy transfer and primary photochemistry in the phycobilisome-photosystem II assembly of Synechocystis sp. PCC 6803. AB - The PsbU subunit of photosystem II (PSII) is one of three extrinsic polypeptides associated with stabilizing the oxygen evolving machinery of photosynthesis in cyanobacteria. We investigated the influence of PsbU on excitation energy transfer and primary photochemistry by spectroscopic analysis of a PsbU-less (or deltaPsbU) mutant. The absence of PsbU was found to have multiple effects on the excited state dynamics of the phycobilisome and PSII. DeltaPsbU cells exhibited decreased variable fluorescence when excited with light absorbed primarily by allophycocyanin but not when excited with light absorbed primarily by chlorophyll a. Fluorescence emission spectra at 77 K showed evidence for impaired energy transfer from the allophycocyanin terminal phycobilisome emitters to PSII. Picosecond fluorescence decay kinetics revealed changes in both allophycocyanin and PSII associated decay components. These changes were consistent with a decrease in the coupling of phycobilisomes to PSII and an increase in the number of closed PSII reaction centers in the dark-adapted deltaPsbU mutant. Our results are consistent with the assumption that PsbU stabilizes both energy transfer and electron transport in the PBS/PSII assembly. PMID- 16363808 TI - Characterization of the redox and metal binding activity of BsSco, a protein implicated in the assembly of cytochrome c oxidase. AB - Members of the Sco protein family are implicated in the assembly of the respiratory complex cytochrome c oxidase. Several possible roles have been proposed for Sco: a copper delivery agent, a site-specific thiol reductase, and an indicator of cellular redox status. Two cysteine residues (C45 and C49) in the sequence CXXXCP and a histidine (H135) approximately 90 residues toward the C terminus are conserved in Sco from bacteria, yeast, and humans. The soluble domain of Sco has a thioredoxin fold that is suggestive of redox activity for this protein. We have characterized the soluble domain of the Sco protein from Bacillus subtilis (i.e., sBsSco) for its redox reactivity and metal binding capacity. In oxidized sBsSco, the cysteines are present as an intramolecular disulfide. Oxidized sBsSco does not bind metal, but can be reduced in vitro to a metal-binding form. Reduction of the disulfide in sBsSco is accompanied by increased intrinsic fluorescence. The reducibility of the cystine is unchanged when the conserved histidine is mutated to alanine. Tight binding by reduced sBsSco is observed for Cu(II) by electronic absorption, intrinsic fluorescence, and EPR spectroscopies, and isothermal titration calorimetry with an observed stoichiometry of one Cu(II) ion per sBsSco and a KD of approximately 50 nM. Tight binding of Cu(I) and Ag(I) is observed by quenching of intrinsic tryptophan fluorescence. Cobalt(II) exhibits weak binding, whereas Ni(II) and Zn(II) do not appear to bind. The high-affinity binding of metals by BsSco is triggered by its redox state, and this property could be important for its function in vivo. PMID- 16363809 TI - Interstrand cross-link induction by UV radiation in bromodeoxyuridine-substituted DNA: dependence on DNA conformation. AB - DNA interstrand cross-links (ICL) can be induced both by natural products (e.g., psoralens with UVA) and by chemical agents, some of which are used in chemotherapy (e.g., Carboplatin and mitomycin C). Here, we report the formation of ICL by UV radiation in brominated DNA, but only for very specific conformations. The quantum yields for strand break and cross-link formation depend on the wavelength with a maximum near 280 nm. It is known that the photosensitization of DNA by bromodeoxyuridine (BrdUrd) results mainly from the electron affinity of bromine, leading to the irreversible formation of 2' deoxyuridin-5-yl radicals (dUrd*) upon the addition of an electron from an adjacent adenosine. It is well documented that the photolytic loss of the bromine atom is greatly suppressed in single-stranded DNA versus that in double-stranded DNA. To study this behavior, we have used two models of BrdUrd-mediated sensitization: one consists of a DNA duplex containing a bulge, formed by five mismatched bases, including the BrdUrd, and the other consists of completely duplex DNA. UV irradiation induces much higher levels of single-strand breaks (ssb) in the completely duplex DNA at the BrdUrd site compared to the DNA with a bulge. However, in completely duplex DNA, ssb appear only in the brominated strand, whereas in the bulged duplex DNA, ssb occur on both strands. Most importantly, we also observe formation of interstrand cross-links in bulged duplex DNA in the BrdUrd region. Thus, we propose that UV irradiation of cells containing BrdUrd incorporated randomly into duplex DNA will create many ssb, whereas BrdUrd present in DNA bulges or open regions in double-stranded DNA (transcription bubbles, replication forks) will lead to potentially lethal damage in both strands in the form of ICL. These findings may help explain the potent clinical antiviral activity of IdUrd and BrdUrd (e.g., IdUrd is used to treat eye infections caused by the herpes virus) and suggest that ICL formation may be a very specific probe for identifying single-stranded regions in the DNA of living cells. In addition, this model system provides an excellent means of introducing ICL for studies on their repair and biological consequences. PMID- 16363810 TI - Mechanism of translocation and kinetics of DNA unwinding by the helicase RecG. AB - RecG is a DNA helicase involved in the repair of damage at a replication fork and catalyzes the reversal of the fork to a point beyond the damage in the template strand. It unwinds duplex DNA in reactions that are coupled to ATP hydrolysis. The kinetic mechanism of duplex DNA unwinding by RecG was analyzed using a quantitative fluorescence assay based on the process of contact quenching between Cy3 and Dabcyl groups attached to synthetic three-way DNA junctions. The data show that the protein moves at a rate of 26 bp s(-1) along the duplex DNA during the unwinding process. RecG ATPase activity during translocation indicates a constant rate of 7.6 s(-1), measured using a fluorescent phosphate sensor, MDCC PBP. These two rates imply a movement of approximately 3 bp per ATP hydrolyzed. We demonstrate in several trapping experiments that RecG remains attached to DNA after translocation to the end of the arm of the synthetic DNA junction. ATPase activity continues after translocation is complete. Dissociation of RecG from the product DNA occurs only very slowly, suggesting strong interactions between them. The data support the idea that interactions of the duplex template arm with the protein are the major sites of binding and production of translocation. PMID- 16363811 TI - Parallels in rRNA processing: conserved features in the processing of the internal transcribed spacer 1 in the pre-rRNA from Schizosaccharomyces pombe. AB - Despite the large differences in their length and nucleotide composition, comparative analyses of the internal transcribed spacer 1 (ITS1) of widely divergent eukaryotes have suggested a simple core structure consisting of a central extended hairpin and lesser hairpin structures at the maturing junctions [Lalev, A. I., and Nazar, R. N. (1998) J. Mol. Biol. 284, 1341-1351]. In this study, the ITS1 in the pre-rRNA transcripts of Schizosaccharomyces pombe cells was examined with respect to structural features that underlie rRNA maturation. When plasmid-associated rRNA genes were expressed in vivo, a deletion of any major hairpin structure significantly reduced or eliminated both small and large subunit RNAs. Only changes in the central extended hairpin or junction regions, however, entirely eliminated plasmid-derived RNAs or resulted in elevated precursor levels. Structure-disrupting base substitutions within the RAC protein complex binding site in the extended hairpin indicated that the secondary structure was critical for rRNA maturation; composition or other changes with respect to the binding site had only modest effects. A similar disruption at the junction with the 18S rRNA also had striking effects on rRNA maturation, including a highly elevated level of unprocessed precursor and a surprisingly critical effect on 5.8S rRNA production. As previously observed with the 3' external transcribed spacer, the results are consistent with a maturation mechanism in which an initial cleavage in the 5' junction region may be directed by the RAC protein complex. Although not critical to rRNA processing, analyses of termini based on S1 nuclease protection as well as cleavage studies, in vitro, with Pac1 ribonuclease raise the possibility that in eukaryotes, as previously observed in bacteria, the RNase III homologues normally initiate the separation of the subunit RNAs. PMID- 16363812 TI - Hydration changes in the association of Hoechst 33258 with DNA. AB - The role of water in the interaction of Hoechst 33258 with the minor groove binding site of the (AATT)2 sequence was investigated using calorimetric and equilibrium constant measurements. Using isothermal titration calorimetry measurements, the heat capacity change for the reaction is -256 +/- 10 cal/(K mol of Hoechst). Comparison with the heat capacity changes based on area models supports the expulsion of water from the interface of the Hoechst-DNA complex. To further consider the role of water, the osmotic stress method was used to determine if the Hoechst association with DNA was coupled with hydration changes. Using four osmolytes with varying molecular weights and chemical properties, the Hoechst affinity for DNA decreases with increasing osmolyte concentration. From the dependence of the equilibrium constant on the solution osmolality, 60 +/- 13 waters are acquired in the complex relative to the reactants. It is proposed that the osmotic stress technique is measuring weakly bound waters that are not measured via the heat capacity changes. PMID- 16363813 TI - Efficient thrombin generation requires molecular phosphatidylserine, not a membrane surface. AB - Activation of prothrombin to thrombin is catalyzed by a "prothrombinase" complex, traditionally viewed as factor X(a) (FX(a)) in complex with factor V(a) (FV(a)) on a phosphatidylserine (PS)-containing membrane surface, which is widely regarded as required for efficient activation. Activation involves cleavage of two peptide bonds and proceeds via one of two released intermediates or through "channeling" (activation without the release of an intermediate). We ask here whether the PS molecule itself and not the membrane surface is sufficient to produce the fully active human "prothrombinase" complex in solution. Both FX(a) and FV(a) bind soluble dicaproyl-phosphatidylserine (C6PS). In the presence of sufficient C6PS to saturate both FX(a) and FV(a2) (light isoform of FV(a)), these proteins form a tight (Kd = 0.6 +/- 0.09 nM at 37 degrees C) soluble complex. Complex assembly occurs well below the critical micelle concentration of C6PS, as established in the presence of the proteins by quasi-elastic light scattering and pyrene fluorescence. Ferguson analysis of native gels shows that the complex migrates with an apparent molecular mass only slightly larger than that expected for one FX(a) and one FV(a2), further ruling out complex assembly on C6PS micelles. Human prothrombin activation by this complex occurs at nearly the same overall rate (2.2 x 10(8) M(-1) s(-1)) and via the same reaction pathway (50-60% channeling, with the rest via the meizothrombin intermediate) as the activation catalyzed by a complex assembled on PS-containing membranes (4.4 x 10(8) M(-1) s( 1)). These results question the accepted role of PS membranes as providing "dimensionality reduction" and favor a regulatory role for platelet-membrane exposed PS. PMID- 16363814 TI - What makes the bioactive lipids phosphatidic acid and lysophosphatidic acid so special? AB - Phosphatidic acid and lysophosphatidic acid are minor but important anionic bioactive lipids involved in a number of key cellular processes, yet these molecules have a simple phosphate headgroup. To find out what is so special about these lipids, we determined the ionization behavior of phosphatidic acid (PA) and lysophosphatidic acid (LPA) in extended (flat) mixed lipid bilayers using magic angle spinning 31P NMR. Our data show two surprising results. First, despite identical phosphomonoester headgroups, LPA carries more negative charge than PA when present in a phosphatidylcholine bilayer. Dehydroxy-LPA [1-oleoyl-3 (phosphoryl)propanediol] behaves in a manner identical to that of PA, indicating that the difference in negative charge between LPA and PA is caused by the hydroxyl on the glycerol backbone of LPA and its interaction with the phosphomonoester headgroup. Second, deprotonation of phosphatidic acid and lysophosphatidic acid was found to be strongly stimulated by the inclusion of phosphatidylethanolamine in the bilayer, indicating that lipid headgroup charge depends on local lipid composition and will vary between the different subcellular locations of (L)PA. Our findings can be understood in terms of a hydrogen bond formed within the phosphomonoester headgroup of (L)PA and its destabilization by competing intra- or intermolecular hydrogen bonds. We propose that this hydrogen bonding property of (L)PA is involved in the various cellular functions of these lipids. PMID- 16363815 TI - The cytoplasmic domains of phospholamban and phospholemman associate with phospholipid membrane surfaces. AB - Phospholamban (PLB) and phospholemman (PLM, also called FXYD1) are small transmembrane proteins that interact with P-type ATPases and regulate ion transport in cardiac cells and other tissues. This work has investigated the hypothesis that the cytoplasmic domains of PLB and PLM, when not interacting with their regulatory targets, are stabilized through associations with the surface of the phospholipid membrane. Peptides representing the 35 C-terminal cytoplasmic residues of PLM (PLM(37-72)), the 23 N-terminal cytoplasmic residues of PLB (PLB(1-23)), and the same sequence phosphorylated at Ser-16 (P-PLB(1-23)) were synthesized to examine their interactions with model membranes composed of zwitterionic phosphatidylcholine (PC) lipids alone or in admixture with anionic phosphatidylglycerol (PG) lipids. Wide-line 2H NMR spectra of PC/PG membranes, with PC deuterated in the choline moiety, indicated that all three peptides interacted with the membrane surface and perturbed the orientation of the choline headgroups. Fluorescence and 31P magic-angle spinning (MAS) NMR measurements indicated that PLB(1-23) and P-PLB(1-23) had a higher affinity for PC/PG membranes, which carry an overall negative surface charge, than for PC membranes, which have no net surface charge. The 31P MAS NMR spectra of the PC/PG membranes in the presence of PLM(37-72), PLB(1-23), and P-PLB(1-23) indicated that all three peptides induced clustering of the lipids into PC-enriched and PG-enriched regions. These findings support the theory that the cytoplasmic domains of PLB and PLM are stabilized by interacting with lipid headgroups at the membrane surface, and it is speculated that such interactions may modulate the functional properties of biological membranes. PMID- 16363816 TI - Amyloid fibril formation by bovine milk kappa-casein and its inhibition by the molecular chaperones alphaS- and beta-casein. AB - Caseins are a unique and diverse group of proteins present in bovine milk. While their function is presumed to be primarily nutritional, caseins have a remarkable ability to stabilize proteins, i.e., to inhibit protein aggregation and precipitation, that is comparable to molecular chaperones of the small heat-shock protein (sHsp) family. Additionally, sHsps have been shown to inhibit the formation of amyloid fibrils. This study investigated (i) the fibril-forming propensities of casein proteins and their mixture, sodium caseinate, and (ii) the ability of caseins to prevent in vitro fibril formation by kappa-casein. Transmission electron microscopy (TEM) and X-ray fiber diffraction data demonstrated that kappa-casein readily forms amyloid fibrils at 37 degrees C particularly following reduction of its disulfide bonds. The time-dependent increase in thioflavin T fluorescence observed for reduced and nonreduced kappa casein at 37 degrees C was suppressed by stoichiometric amounts of alphaS- and beta-casein and by the hydrophobic dye 8-anilino-1-naphthalene sulfonate; the inhibition of kappa-casein fibril formation under these conditions was verified by TEM. Our findings suggest that alphaS- and beta-casein are potent inhibitors of kappa-casein fibril formation and may prevent large-scale fibril formation in vivo. Casein proteins may therefore play a preventative role in the development of corpora amylacea, a disorder associated with the accumulation of amyloid deposits in mammary tissue. PMID- 16363817 TI - Synthesis, characterization, and cytotoxicity of a novel highly charged trinuclear platinum compound. Enhancement of cellular uptake with charge. AB - Charge delocalization (6+ to 8+) in "noncovalent" linear trinuclear platinum complexes produces compounds with cytotoxicity in some cases equivalent to cisplatin. The cellular uptake of a novel 8+ compound is greater than that of neutral cisplatin as well as other multinuclear Pt compounds. PMID- 16363818 TI - Unusual, bifurcated photoreactivity of a rhenium(I) carbonyl complex of triethynylphosphine. AB - Preparations of the first metal complexes of triethynylphosphine (TEP) are described. They are of the type fac-Re(bpy)(CO)(3)(TEP)(+) (1) and cis,trans [Re(bpy)(CO)(2)(TEP)L](n)(+) (CH(3)CN, n = 1, complex 2; Cl, n = 0, complex 3), where bpy is 2,2'-bipyridine. Complex 1 displays unusual photochemical behavior compared to analogous fac-[Re(bpy)(CO)(3)(PR(3))](+) complexes in that it emits from a state that has pi-pi* character but undergoes competitive photosubstitution of both TEP and CO. Density functional theory (DFT)/time dependent DFT calculations predict that the lowest emitting state should, in fact, have pi-pi* character. PMID- 16363819 TI - Synthesis and structural analysis of the first nanosized platinum-gold carbonyl/phosphine cluster, Pt13[Au2(PPh3)2]2(CO)10(PPh3)4, containing a Pt centered [Ph3PAu-AuPPh3]-capped icosahedral Pt12 cage. AB - The preparation and molecular structure of the initial nanosized platinum-gold carbonyl cluster, Pt(13)[Au(2)(PPh(3))(2)](2)(CO)(10)(PPh(3))(4) (1), are described. A comparative analysis reveals its pseudo-D(2)(h) geometry, consisting of a centered Pt(13) icosahedron encapsulated by two centrosymmetrically related bidentate [Ph(3)PAu-AuPPh(3)]-capped ligands along with 4 PR(3) and 10 CO ligands, to be remarkably similar to that of the previously reported Pt(17)(mu(2) CO)(4)(CO)(8)(PEt(3))(8) (2). Reformulation of 2 as Pt(13)[(PtPEt(3))(2)(mu(2) CO)](2)(CO)(10)(PEt(3))(4) emphasizes the steric/electronic resemblance of the bulky-sized bidentate [Ph(3)PAu-AuPPh(3)] and [(PtPEt(3))(2)(mu(2)-CO)] capping ligands in 1 and 2, respectively, as well as their identical electron counts of 162 cluster valence electrons for a centered Pt(13) icosahedron. We hypothesize that analogous steric effects of their ligand polyhedra in 1 and 2 play a crucial role along with electronic effects in the formation and stabilization of these two nanosized clusters that contain an otherwise unknown centered icosahedron of platinum atoms. PMID- 16363820 TI - A favorable case where an experimental electron density analysis offers a lead for understanding a specific fluxional process observed in solution. AB - A topological analysis of the electron density in the ketene complex (eta(5) MeC(5)H(4))(CO)(2)Mn[eta(2)-O=C=C((mu-eta(2)-CCPh)Co(2)(CO)(6))Ph] indicates a predisposition for the carbene component of the ketene ligand to bind the neighboring C atom of the adjacent CO ligand. PMID- 16363821 TI - [M(R-dmet)2] bis(1,2-dithiolenes): a promising new class intermediate between [M(dmit)2] and [M(R,R'-timdt)2] (M = Ni, Pd, Pt). AB - We report the first examples of metal dithiolenes belonging to the class [M(R dmet)(2)] [R-dmet = formally monoreduced N-substituted thiazolidine-2,4,5 trithione; R = Et, M = Ni (1), Pd (2), Pt (3)]. A comparative spectroscopic, electrochemical, and density functional theory theoretical investigation indicates that [M(R-dmet)(2)] complexes show features intermediate between those of the dithiolenes belonging to the previously reported classes [M(R,R' timdt)(2)] and [M(dmit)(2)] (R,R'-timdt = formally monoreduced N,N'-disubstituted imidazolidine-2,4,5-trithione; dmit = 2-thioxo-1,3-dithiole-4,5-dithiolato). UV vis-near-IR spectroscopy and cyclic voltammetry/differential pulsed voltammetry measurements performed on 1 and 3 proved that the new dithiolenes are stable as neutral, monoanionic, and bianionic species and feature a near-IR electrochromic absorption falling at about 1000 and 1250 nm for neutral and monoanionic species, respectively. PMID- 16363822 TI - Why are carborane acids so acidic? An electrostatic interpretation of Bronsted acid strengths. AB - Acidity of Bronsted acids is explained in terms of the electrostatic potentials of the corresponding conjugate bases. The electrostatic potential distribution on the zero-flux surface of the strongest isolable carborane anions is seen to provide a good measure of their acidities. Increasing value of the lowest minimum in the electrostatic potential is observed to be a signature of increasing acidity. PMID- 16363823 TI - Methoxide of an Anderson-type polyoxometalate and its conversion to a new type of species, [IMo9O32(OH)(OH2)3]4-. AB - A totally new type of polyoxometalate, [IMo(9)O(32)(OH)(OH(2))(3)](4)(-), has been synthesized by reacting [IMo(6)O(22)(OMe)(2)](3)(-) with water. The [IMo(9)O(32)(OH)(OH(2))(3)](4)(-) anion further transforms into [(IMo(7)O(26))(2)](6)(-), a molecular oxide that has a rutile core, in dry acetonitrile, while it stays intact for several hours in wet acetonitrile. PMID- 16363824 TI - Solid-state photochromic device based on nanocrystalline TiO2 functionalized with electron donor-acceptor species. AB - In this Communication, we report on a new type of solid-state photochromic device, S-TiO(2)-A, which is based on electron-transfer processes involving a molecular sensitizer (S) and an electron acceptor (A) coadsorbed onto the surface of nanocrystalline TiO(2). The electron-transfer process is mediated by the semiconductor conduction band and leads to a long-lived charge-separated state S(+)/A(-). The lifetime of this state can be controlled by oxygen diffusion through a polymeric coating deposited on the device. PMID- 16363825 TI - Reduction of the host cationic framework charge by isoelectronic substitution: synthesis and structure of Hg7Ag2P8X6 (X = Br, I) and Hg6Ag4P8Br6. AB - The first compounds, Hg(7)Ag(2)P(8)X(6) (X = Br, I) and Hg(6)Ag(4)P(8)Br(6), featuring the partial isoelectronic substitution of Hg(2+) for Ag(1+) in mercury pnicogen frameworks have been obtained and structurally characterized. The new compounds are the supramolecular assemblies built of the covalently bonded metal pnicogen frameworks trapping guests of different complexity. The frameworks feature the perfect ordering of Hg(2+) and Ag(1+) cations and contain P(2)(4)(-) and P(6)(6)(-) phosphorus clusters. The substitution of Hg(2+) with Ag(1+) leads to the reduction in charge of the host cluster-containing cationic matrix and concomitant replacement of the monatomic X(-) guest by a lesser amount of the AgBr(3)(2)(-) anions. PMID- 16363826 TI - Highly sensitive ammonium tetraazidoaurates(III). AB - The preparation and characterization of selected ammonium and methylammonium tetraazidoaurates(III) are reported. All ammonium salts were shown to be highly explosive materials. The first crystal structure of such an ammonium salt, that of [Me(4)N][Au(N(3))(4)], features polymeric units of the anion, which are linked by weak Au...Au interactions. PMID- 16363827 TI - Oxygen binding of water-soluble cobalt porphyrins in aqueous solution. AB - Water-soluble cobalt porphyrin 1Co and imidazole ligand 2 were synthesized. 1Co binds dioxygen in the presence of imidazole ligand 2 in aqueous solution. The formation of the oxygen adduct 2-1Co(O(2)) was studied using UV-vis and EPR spectroscopy. The impact of pH on the kinetic stability of the oxygen adduct was examined. PMID- 16363828 TI - mer-[Fe(pcq)(CN)3]-: a novel cyanide-containing building block and its application to assembling cyanide-bridged trinuclear FeIII2MnII complexes [pcq- = 8-(pyridine-2-carboxamido)quinoline anion]. AB - A new cyanide-containing building block K[Fe(pcq)(CN)(3)] [1; pcq(-) = 8 (pyridine-2-carboxamido)quinoline anion] containing a low-spin Fe(III) center with three cyanide groups in a meridional arrangement has been successfully designed and synthesized. Three cyanide-bridged trinuclear Fe(III)(2)Mn(II) complexes, [Fe(pcq)(CN)(3)](2)[Mn(CH(3)OH)(2)(H(2)O)(2)].2H(2)O (2), [Fe(pcq)(CN)(3)](2)[Mn(bipy)(2)].CH(3)OH.2H(2)O (3), and [Fe(pcq)(CN)(3)](2)[Mn(phen)(2)].CH(3)OH.2H(2)O (4), have been synthesized and structurally characterized. The magnetic susceptibilities of the three heterometallic complexes have been investigated. PMID- 16363829 TI - Secondary bonding interactions observed in two arsenic thiolate complexes. AB - Treatment of N-(2-mercaptoethyl)-1,8-naphthalimide (HL) with stoichiometric amounts of AsCl(3) and base affords AsL(2)Cl and AsL(3) complexes stabilized in part by secondary As...O bonding interactions. PMID- 16363830 TI - Synthesis and structural characterization of M(PMe3)3(O2CR)2(OH2)H2 (M = Mo, W): aqua-hydride complexes of molybdenum and tungsten. AB - Mo(PMe(3))(6) and W(PMe(3))(4)(eta(2)-CH(2)PMe(2))H undergo oxidative addition of the O-H bond of RCO(2)H to yield sequentially M(PMe(3))(4)(eta(2)-O(2)CR)H and M(PMe(3))(3)(eta(2)-O(2)CR)(eta(1)-O(2)CR)H(2) (M = Mo and R = Ph, Bu(t); M = W and R = Bu(t)). One of the oxygen donors of the bidentate carboxylate ligand may be displaced by H(2)O to give rare examples of aqua-dihydride complexes, M(PMe(3))(3)(eta(1)-O(2)CR)(2)(OH(2))H(2), in which the coordinated water molecule is hydrogen-bonded to both carboxylate ligands. PMID- 16363831 TI - Theoretical study of catalytic dinitrogen reduction under mild conditions. AB - Density functional theory results on key steps of Schrock's catalytic cycle are presented. The quantum chemical modeling of the dinitrogen-reducing reaction steps is conducted without simplifying the bulky HIPT [HIPT = 3,5-(2,4,6 iPr(3)C(6)H(2))(2)C(6)H(3)] substituents at the triamidoamine ligand. PMID- 16363832 TI - Metal-metal bond or isolated metal centers? Interaction of Hg(CN)2 with square planar transition metal cyanides. AB - Three adducts have been prepared from Hg(CN)(2) and square planar M(II)(CN)(4)(2)(-) transition metal cyanides (M = Pt, Pd, or Ni, with d(8) electron shell) as solids. The structure of the compounds K(2)PtHg(CN)(6).2H(2)O (1), Na(2)PdHg(CN)(6).2H(2)O (2), and K(2)NiHg(CN)(6).2H(2)O (3) have been studied by single-crystal X-ray diffraction, XPS, Raman spectroscopy, and luminescence spectroscopy in the solid state. The structure of K(2)PtHg(CN)(6).2H(2)O consists of one-dimensional wires. No CN(-) bridges occur between the heterometallic centers. The wires are strictly linear, and the Pt(II) and Hg(II) centers alternate. The distance d(Hg)(-)(Pt) is relatively short, 3.460 A. Time-resolved luminescence spectra indicate that Hg(CN)(2) units incorporated into the structure act as electron traps and shorten the lifetime of both the short-lived and longer-lived exited states in 1 compared to K(2)[Pt(CN)(4)].2H(2)O. The structures of Na(2)PdHg(CN)(6).2H(2)O and K(2)NiHg(CN)(6).2H(2)O can be considered as double salts; the lack of heterometallophilic interaction between the remote Hg(II) and Pd(II) atoms, d(Hg)(-)(Pd) = 4.92 A, and Hg(II) and Ni(II) atoms, d(Hg)(-)(Ni) = 4.61 A, is apparent. Electron binding energy values of the metallic centers measured by XPS show that there is no electron transfer between the metal ions in the three adducts. In solution, experimental findings clearly indicate the lack of metal metal bond formation in all studied Hg(II)-CN(-)-M(II)(CN)(4)(2)(-) systems (M = Pt, Pd, or Ni). PMID- 16363833 TI - MXAN analysis of the XANES energy region of a mononuclear copper complex: applications to bioinorganic systems. AB - The near edge XAS spectra of the mononuclear copper complex [Cu(TMPA)(OH(2))](ClO(4))(2) (1) have been simulated using the multiple scattering edge simulation package MXAN (or Minuit XANes). These simulations, which employ the muffin-tin (MT) approximation, have been compared to simulations generated using the finite-difference method (FDM) to evaluate the effect of MT corrections. The sensitivity of the MXAN method was tested using structural models that included several different variations on the bond angles and bond distances for the first-shell atoms of 1. The sensitivity to small structural changes was also evaluated by comparing MXAN simulations of 1 and of structurally modified [Cu(TMPA)(L)](n)(+) complexes [where L = -O-(F(8)TPP)Fe(III), -F, OPO(2)(O-p-nitrophenyl)Zn(II)(TMPA), and -NCMe] to the experimental data. The accuracy of the bond distances obtained from the MXAN simulations was then examined by comparison to the metrics of the crystal structures. The results show that MXAN can successfully extract geometric information from the edge structure of an XAS spectrum. The systematic application of MXAN to 1 indicates that this approach is sensitive to small structural changes in the molecule that are manifested in the XAS edge spectrum. These results represent the first step toward the application of this methodology to bioinorganic and biological systems. PMID- 16363834 TI - ((Fluoroformyl)imido)sulfuryl difluoride, FC(O)N=S(O)F2: structural, conformational, and configurational properties in the gaseous and condensed phases. AB - Structural, conformational, and configurational properties of the gaseous molecule ((fluoroformyl)imido)sulfuryl difluoride, FC(O)N=S(O)F(2), have been studied by vibrational spectroscopy (IR (gas) and Raman (liquid)) and quantum chemical calculations (HF, MP2, and B3LYP with 6-31+G* and 6-311+G* basis sets); in addition, the solid-state structure has been determined by X-ray crystallography. FC(O)N=S(O)F(2) exists in the gas phase as a mixture of a favored antiperiplanar-synperiplanar form (the S=O double bond antiperiplanar with respect to the C-N single bond, and the C=O group synperiplanar with respect to the S=N double bond) in equilibrium with less abundant antiperiplanar antiperiplanar, synclinal-synperiplanar, and synclinal-antiperiplanar structures. The crystalline solid at 163 K (monoclinic, P2(1)/c, a = 5.1323(7) A, b = 15.942(2) A, c = 16.798(2) A, beta = 95.974(3) degrees , Z = 12) consists of three similar antiperiplanar-synperiplanar forms. PMID- 16363835 TI - Luminescence ranging from red to blue: a series of copper(I)-halide complexes having rhombic {Cu2(mu-X)2} (X = Br and I) units with N-heteroaromatic ligands. AB - A series of Cu(I) complexes formulated as [Cu(2)(mu-X)(2)(PPh(3))(L)(n)] were prepared with various mono- and bidentate N-heteroaromatic ligands (X = Br, I; L = 4,4'-bipyridine, pyrazine, pyrimidine, 1,5-naphthyridine, 1,6-naphthyridine, quinazoline, N,N-dimethyl-4-aminopyridine, 3-benzoylpyridine, 4-benzoylpyridine; n = 1, 2). Single-crystal structure analyses revealed that all the complexes have planar {Cu(2)X(2)} units. Whereas those with monodentate N-heteroaromatic ligands afforded discrete dinuclear complexes, bidentate ligands formed infinite chain complexes with the ligands bridging the dimeric units. The long Cu...Cu distances (2.872-3.303 A) observed in these complexes indicated no substantial interaction between the two Cu(I) ions. The complexes showed strong emission at room temperature as well as at 80 K in the solid state. The emission spectra and lifetimes in the microsecond range were measured at room temperature and at 80 K. The emissions of the complexes varied from red to blue by the systematic selection of the N-heteroaromatic ligands (lambda(em)(max): 450 nm (L = N,N dimethyl-4-aminopyridine) to 707 nm (L = pyrazine)), and were assigned to metal to-ligand charge-transfer (MLCT) excited states with some mixing of the halide-to ligand (XL) CT characters. The emission energies were successfully correlated with the reduction potentials of the coordinated N-heteroaromatic ligands, which were estimated by applying a simple modification based on the calculated stabilization energies of the ligands by protonation. PMID- 16363836 TI - Synthesis, structure, and magnetic properties of Sr2NiOsO6 and Ca2NiOsO6: two new osmium-containing double perovskites. AB - Two new double perovskite oxides, Ca(2)NiOsO(6) and Sr(2)NiOsO(6), have been prepared as polycrystalline powders by solid state synthesis. The two oxides were structurally characterized by variable-temperature powder neutron diffraction. Ca(2)NiOsO(6) was found to adopt a monoclinic structure (P2(1)/n), while Sr(2)NiOsO(6) was found to be tetragonal (I4/m). Magnetic susceptibility measurements indicate that Ca(2)NiOsO(6) orders in a canted antiferromagnetic state at about 175 K while Sr(2)NiOsO(6) orders antiferromagnetically at about 50 K. PMID- 16363837 TI - A novel diamine adduct of zinc bis(2-thenoyl-trifluoroacetonate) as a promising precursor for MOCVD of zinc oxide films. AB - A novel diamine (N,N,N',N'-tetramethyletilendiamine) adduct of zinc bis(2-thenoyl trifluoroacetonate) has been synthesized in a single-step reaction. Single crystal X-ray diffraction studies of Zn(tta)(2).tmeda provide evidence of a mononuclear structure with a six-coordinated zinc ion. The thermal behavior of this adduct points to mass-transport properties suitable for its application to MOCVD processes. This novel compound has been successfully applied as a precursor for the deposition of ZnO films on (100) Si and quartz substrates. The good quality of the deposited films indicates that the adduct is a very attractive precursor for MOCVD applications. PMID- 16363838 TI - Controlling emission energy, self-quenching, and excimer formation in highly luminescent N--C--N-coordinated platinum(II) complexes. AB - A series of cyclometalated platinum(II) complexes have been prepared, [PtL(n)Cl], containing N--C--N-coordinating, terdentate ligands based on 1,3-dipyridylbenzene (HL(1)), incorporating aryl substituents at the central 5 position of the ligand. All of the new complexes are intensely luminescent in a degassed solution at 298 K (phi = 0.46-0.65 in CH(2)Cl(2)) with lifetimes in the microsecond range (7.9 20.5 micros). The introduction of the aryl substituents leads to a red shift in the lowest-energy, intense charge-transfer absorption band compared to [PtL(1)Cl] (401 nm in CH(2)Cl(2)), in the order H < mesityl < 2-pyridyl < 4-tolyl < 4 biphenylyl < 2-thienyl < 4-(dimethylamino)phenyl (431 nm in CH(2)Cl(2)), which correlates with the decreasing order of oxidation potentials. A similar order is also observed in the emission maxima, ranging from 491 nm for [PtL(1)Cl] to 588 nm for the 4-(dimethylamino)phenyl-substituted complex. The emission spectra of all of the complexes, except for the amino-substituted compound, are highly structured in a dilute solution in CH(2)Cl(2), and the emission is assigned to excited states of primarily (3)LC (ligand-centered) character. At higher concentrations, self-quenching accompanied by structureless excimer emission centered at 700 nm is observed, but the aryl groups attenuate the self-quenching compared to the parent compound [PtL(1)Cl], particularly for the most sterically hindered mesityl complex. The introduction of the strongly electron-donating 4 dimethylamino substituent leads to a switch in the nature of the lowest-energy excited state from (3)LC to one of primarily intraligand charge-transfer (ILCT) character in CH(2)Cl(2): this complex displays a structureless and much broader emission band than the other compounds and a high degree of positive solvatochromism. No excimer emission is observed in CH(2)Cl(2), and self quenching is an order of magnitude lower than that for the other complexes. However, in nonpolar solvents such as CCl(4), the ILCT state is destabilized, such that the (3)LC remains the lowest-energy excited state. Reversible switching between the ILCT and (3)LC states can also be achieved in a CH(2)Cl(2) solution by protonation of the amine, with an accompanying large change in the emission maxima of >100 nm. The X-ray structures of the biphenylyl- and methyl-substituted complexes are reported, together with those of the 2-pyridyl- and mesityl substituted ligands and the key synthetic intermediate 1-bromo-3,5-di(2 pyridyl)benzene. PMID- 16363840 TI - Syntheses, crystal structures, and spectroscopic and magnetic properties of [Mn2III(H2L1)(Cl4Cat)4.2H2O] and [Mn2III(H2L2)(Cl4Cat)4.2CH3CN.2H2O]infinity: temperature-dependent valence tautomerism in solution. AB - The synthesis, X-ray data, and electronic structures of two manganese(III) 1D polymers ligated by tetrachlorocatechol, [Mn(2)(III)(H(2)L(1))(Cl(4)Cat)(4).2H(2)O](infinity) (1) and [Mn(2)(III)(H(2)L(2))(Cl(4)Cat)(4).2CH(3)CN.2H(2)O](infinity) (2), are reported. The electronic structures of the complexes have been determined by UV-vis-near IR, IR, electron paramagnetic resonance (EPR), and magnetic susceptibility measurements. Both 1 and 2 are air stable in the solid state and in solution, unlike most of the previously reported o-quinone-chelated transition-metal complexes. Electronic spectroscopy exhibits a strong near-IR band near 1900 nm for both, suggesting the presence of a mixed-valence semiquinone-catecholate oxidation state of the catechol ligands, Mn(2)(III)(Cl(4)Cat)(2)(Cl(4)SQ)(2), together with the pure catecholate forms. The presence of this isomer was further supported by EPR and magnetic susceptibility measurements. The complexes undergo intramolecular electron transfer (valence tautomerism) upon an increase of the temperature involving the equilibrium Mn(2)(III)(Cl(4)Cat)(2)(Cl(4)SQ)(2) <==> Mn(2)(II)(Cl(4)SQ)(4). This phenomenon is reversible and is studied in solution using UV-vis-near-IR spectroscopy. PMID- 16363839 TI - Electronic structure study of seven-coordinate first-row transition metal complexes derived from 1,10-diaza-15-crown-5: a successful marriage of theory with experiment. AB - A detailed study of the electronic structure of seven-coordinate Mn(II), Co(II), and Ni(II) complexes with the lariat ether N,N'-bis(2-aminobenzyl)-1,10-diaza-15 crown-5 (L(1)) is presented. These complexes represent new examples of structurally characterized seven-coordinate (pentagonal bipyramidal) complexes for the Mn(II), Co(II), and Ni(II) ions. The X-ray crystal structures of the Mn(II) and Co(II) complexes show C(2) symmetries for the [M(L(1))](2+) cations, whereas the structures of the Ni(II) complexes show a more distorted coordination environment. The magnetic properties of the Mn(II) complex display a characteristic Curie law, whereas those of the Co(II) and Ni(II) ions show the occurrence of zero-field splitting of the S = 3/2 and 1 ground states, respectively. Geometry optimizations of the [M(L(1))](2+) systems (M = Mn, Co, or Ni) at the DFT (B3LYP) level of theory provide theoretical structures in good agreement with the experimental data. Electronic structure calculations predict a similar ordering of the metal-based beta spin frontier MO for the Mn(II) and Co(II) complexes. This particular ordering of the frontier MO leads to a pseudodegenerate ground state for the d(8) Ni(II) ion. The distortion of the C(2) symmetry in [Ni(L(1))](2+) is consistent with a Jahn-Teller effect that removes this pseudodegeneracy. Our electronic structure calculations predict that the binding strength of L(1) should follow the trend Co(II) approximately Mn(II) > Ni(II), in agreement with experimental data obtained from spectrophotometric titrations. PMID- 16363841 TI - Crystal structure, magnetic properties, and 57Fe Mossbauer spectroscopy of the two-dimensional coordination polymers [M(1,2-bis(1,2,4-triazol-4 yl)ethane)2(NCS)2] (MII = Fe, Co). AB - New coordination polymers of the formula [M(btre)(2)(NCS)(2)] (btre = 1,2 bis(1,2,4-triazol-4-yl)ethane; M(II) = Fe, Co) have been synthesized, and their crystal structures have been determined at 293 K by X-ray analysis. The Fe(II) compound (C(7)H(8)FeN(7)S(2)) crystallizes in the monoclinic space group P2(1)/n, a = 12.439(5) A, b = 8.941(2) A, c = 9.321(3) A, beta = 90.88(2) degrees , V = 1036.6(6) A(3), Z = 2, 3791 reflections [I > 3sigma(I)], R(F) = 0.036, wR2 = 0.123. The Co(II) compound is isostructural to the Fe(II) compound. The crystal structure consists of a 2D sheet in which the metal ions are linked by bis monodentate (N1, N1') 1,2,4-triazole ligands. The structure is stabilized by pi bond interactions between two adjacent sheets and by S...S interactions. Temperature-dependent SQUID, (57)Fe Mossbauer, and X-ray diffraction measurements indicate that [Fe(btre)(2)(NCS)(2)] retains a HS ground state upon cooling from 293 K down to 8 K. The surprising absence of spin-crossover behavior for this Fe(II)-1,2,4-triazole polymeric coordination compound that has been confirmed by pressure experiments up to approximately 12 kbar and by light irradiation experiments at 10 K is discussed on the basis of its structural features. Insight into the origin of the cooperative effects of the spin transition in [Fe(btr)(2)(NCS)(2)].H(2)O (btr = 4,4'-bis-1,2,4-triazole) is also given thanks to a re-evaluation of its distortion parameters in the high- and low-spin states. PMID- 16363842 TI - Inorganic-metalorganic hybrids based on copper(II)-monosubstituted Keggin polyanions and dinuclear copper(II)-oxalate complexes. Synthesis, X-ray structural characterization, and magnetic properties. AB - Reaction of in situ generated copper(II)-monosubstituted Keggin polyoxometalates and copper(II)-bipyridine-oxalate complexes in the corresponding alkaline acetate buffer led to the formation of hybrid metal organic-inorganic compounds K(2)[{SiW(11)O(39)Cu(H(2)O)}{Cu(2)(bpy)(2)(H(2)O)(2)(mu-ox)}(2)].14H(2)O (1), K(14)[{SiW(11)O(39)Cu(H(2)O)}{Cu(2)(bpy)(2)(mu ox)}](2)[SiW(11)O(39)Cu(H(2)O)].55H(2)O (2), (NH(4))(4)[{SiW(11)O(39)Cu(H(2)O)}{Cu(2)(bpy)(2)(mu-ox)}].10H(2)O (3), and Rb(4)[{SiW(11)O(39)Cu(H(2)O)}{Cu(2)(bpy)(2)(mu-ox)}].10H(2)O (4). Their structures have been established by single-crystal X-ray diffraction. The main structural feature of these compounds is the presence of copper(II) monosubstituted alpha-Keggin polyoxoanions as inorganic building blocks, on which the mu-oxalatodicopper metalorganic blocks are supported. Compound 1contains the discrete hybrid polyanion [{SiW(11)O(39)Cu(H(2)O)}{Cu(2)(bpy)(2)(H(2)O)(2)(mu ox)}(2)](2)(-), whereas the polymeric hybrid polyanion [{SiW(11)O(39)Cu(H(2)O)}{Cu(2)(bpy)(2)(mu-ox)}(2)](n)(4)(n)(-) gives a monodimensional character to compounds 2-4. Magnetic and EPR results are discussed with respect to the crystal structure of the compounds. DFT calculations on both the [Cu(2)(bpy)(2)(H(2)O)(4)(mu-ox)](2+) cationic complex and the metalorganic blocks have been performed in order to determine the optimized geometry and the magnetic coupling constants, respectively. PMID- 16363843 TI - X-ray diffraction and EXAFS studies of hydroxo-Cu(II) complexes based on a calix[6]arene-N3 ligand: evidence for a mononuclear-dinuclear equilibrium controlled by supramolecular features. AB - The formation of hydroxo complexes based on a calix[6]trisimidazole ligand is described. Deprotonation of the mononuclear Cu(II)-aqua complex gives rise to a dinuclear bis(mu-hydroxo) complex that has been characterized by X-ray diffraction analysis. Spectroscopic studies (EPR and UV-vis), conducted in dichloromethane solutions in the presence of various coordinating cosolvents (DMF, EtOH, or RCN) or with acetamide, revealed the coexistence of a mononuclear hydroxo species. The latter could be trapped by acetic acid to provide an acetato Cu(II) complex that presents close spectroscopic features. An EXAFS study first conducted on the hydroxo-Cu(II) complex led to an excellent fit for the dinuclear core. It then allowed for the characterization of the mononuclear acetato complex with an acetamide guest included in the calixarene cavity. Hence, this study illustrates the flexibility of calixarene-based ligands and the role of the second coordination sphere in the stabilization of acidic or basic Cu(II) complexes. PMID- 16363844 TI - Pyridylporphyrin metallacycles with a slipped cofacial geometry: spectroscopic, X ray, and photophysical characterization. AB - Treatment of the octahedral Ru(II) complex [trans,cis,cis-RuCl(2)(DMSO O)(2)(CO)(2)] with an equimolar amount of 5,10-bis(3'-pyridyl)-15,20 diphenylporphyrin (3'-cis-DPyP) yielded, upon selective replacement of the DMSO ligands, the neutral 2 + 2 metallacycle 2. NMR spectroscopy provided unambiguous evidence that only one highly symmetrical species, in which the two chromophores are held in a slipped cofacial arrangement by the external Ru(II) metal fragments, exists in solution. The unprecedented geometry of 2, and of the fully zincated analogue 2a, were confirmed in the solid state by X-ray structural investigations. The spatial arrangement of the two parallel chromophores in 2, with an interplanar distance of 4.18 A and a lateral offset (center-to-center distance) of 9.82 A, is reminiscent of those of the special pair of bacteriophylls in the reaction centers and of adjacent B850 units in the LH2 light-harvesting antenna systems of photosynthetic bacteria. For comparison, the X-ray structure of the corresponding metallacycle with 4'-cis-DPyP, 1a, is also reported. In 1a, the two porphyrins have an almost perfect coplanar arrangement. The semi-zincated metallacycles 1b and 2b, in which only one of the two chromophores bears an inner zinc atom, were prepared from 1 and 2, respectively, and isolated in pure form. Detailed photophysical investigations of the above porphyrin assemblies were performed. In particular, very fast photoinduced intercomponent energy transfer processes from the zinc porphyrin to the free-base unit were detected in the semi-metalated derivatives 1b and 2b (time constants: 14 and 12 ps, respectively). PMID- 16363845 TI - Between Ni(mnt)2 and Ni(tfd)2 dithiolene complexes: the unsymmetrical 2 (trifluoromethyl)acrylonitrile-1,2-dithiolate and its nickel complexes. AB - A novel 1,2-dithiolate ligand, that is, the 2-(trifluoromethyl)acrylonitrile-1,2 dithiolate, abbreviated here as tfadt, is prepared from the corresponding cyclic dithiocarbonate. This ligand, substituted with both a CN and a CF(3) group, is compared with the well-known maleonitrile- and bis(trifluoromethyl)ethane-1,2 dithiolates. The preparation, electrochemical properties, and X-ray crystal structures of the square-planar nickel complexes, in both their dianionic diamagnetic [Ni(tfadt)(2)](2)(-) and their monoanionic paramagnetic [Ni(tfadt)(2)](*)(-) forms, are reported, as n-Bu(4)N(+), PPh(4)(+), and (18 crown-6)Na(+) salts, respectively. In the [(18-crown-6)Na](2)[Ni(tfadt)(2)] salt, each CN moiety of the [Ni(tfadt)(2)](2)(-) dianion is coordinated to a (18-crown 6)Na(+) cation through a CN...Na interaction [N...Na = 2.481(3) A], affording an "axle with wheels" model where two MeOH molecules act as axle caps. On the other hand, in [(18-crown-6)Na][Ni(tfadt)(2)], each (18-crown-6)Na(+) cation is coordinated on both sides by the CN groups of two monoanionic [Ni(tfadt)(2)](*)( ) complexes with N...Na(+) distances at 2.434(5) and 2.485(4) A, giving rise to heterobimetallic chains with alternating (18-crown-6)Na(+) and [Ni(tfadt)(2)](*)( ) ions. These two examples demonstrate the attractive ability of the CN moieties in the [Ni(tfadt)(2)](2)(-)(,)(*)(-) complexes to coordinate metallic cationic centers. The paramagnetic salts of the anionic [Ni(tfadt)(2)](*)(-) complex follow Curie-type law in the 2-300 K temperature range, indicating the absence of intermolecular magnetic interactions in the solid state. The complexes are found in their trans form in all crystal structures, while density functional theory calculations establish that both forms have essentially the same energy. A cis trans interconversion process is observed by variable-temperature NMR on the dianionic [Ni(tfadt)(2)](2)(-) complex with a coalescence temperature T(c) of 260 K and a free energy of activation of 51-53 kJ mol(-)(1). PMID- 16363846 TI - Fluorocarbon soluble copper(II) carboxylate complexes with nonfluoroponytailed nitrogen ligands as precatalysts for the oxidation of alkenols and alcohols under fluorous biphasic or thermomorphic modes: structural and mechanistic aspects. AB - This fluorous biphasic catalysis (FBC) contribution was focused on the synthesis and characterization of new fluorous soluble R(f)-Cu(II) carboxylate complexes containing nonfluoroponytailed ligands and defines their role as precatalysts for the FBC oxidation of alkenols and alcohols in the presence of 2,2,6,6 tetramethylpiperidine-N-oxyl (TEMPO)/O(2). In this FBC approach, we have utilized the phase-switching technique of Vincent et al. (J. Am. Chem. Soc. 2002, 124, 12942) to solubilize the nonfluoroponytailed ligands, N-1,4,7-Me(3)TACN, 2, and N 1,4,7-pentamethyldiethylenetriamine (PMDETA), 3, by reaction with a fluorous solvent-soluble copper (II) dimeric complex, [Cu({C(8)F(17)(CH(2))(2)}(2)CHCO(2))(2)](2), 1. Moreover, the reaction of nonfluoroponytailed ligands 2 and 3 with 1 afforded new perfluoroheptane-soluble Cu(II) complexes, [Cu({C(8)F(17)(CH(2))(2)}(2)CHCO(2))(2)(2)], 4, and [Cu({C(8)F(17)(CH(2))(2)}(2)CHCO(2))(2) (3)], 5, respectively. The known Cu(II) complex, 1, was further characterized by electron paramagnetic resonance (EPR) spectroscopy confirming its dimeric structure, while 4 and 5 were characterized by elemental analysis, IR, diffuse reflectance UV-vis, and EPR spectroscopy. Furthermore, 1, 4, and 5 were evaluated as precatalysts for alkenol and alcohol oxidation. The oxidation reactions of alkenols and alcohols in the presence of TEMPO/O(2) proceeded under FBC conditions for 1, 4, and 5, but 1-octanol was unreactive under single-phase FBC conditions at 90 degrees C with TEMPO/O(2). The thermomorphic property of 5, soluble in chlorobenzene/toluene at 90 degrees C but insoluble at room temperature, was also evaluated in the selective oxidation of p nitrobenzyl alcohol to p-nitrobenzaldehyde. Plausible mechanisms concerning these FBC/thermomorphic oxidation reactions will be discussed. PMID- 16363847 TI - Cadmium(II) and zinc(II) complexes of pyrrole-appended oxacarbaporphyrin: a side on coordination mode of O-confused carbaporphyrin. AB - A pyrrole adduct of 5,20-diphenyl-10,15-di(p-tolyl)-2-oxa-21-carbaporphyrin [(H,pyr)OCPH]H(2) reacted with sodium ethanolate to yield 5,20-diphenyl-10,15 di(p-tolyl)-3-ethoxy-3-(2'-pyrrol)-2-oxa-21-carbaporphyrin [(EtO,pyr)OCPH]H(2). Subsequently, "true" O-confused oxaporphyrin with a pendant pyrrole ring [(pyr)OCPH]H was formed by the addition of acid to [(EtO,pyr)OCPH]H(2), which triggered an ethanol elimination. In the course of this process, the tetrahedral trigonal rearrangements originated at the C(3) atom. Insertion of zinc(II), cadmium(II), and nickel(II) into [(pyr)OCPH]H yielded [(pyr)OCPH]Zn(II)Cl, [(pyr)OCPH]Cd(II)Cl, and [(pyr)OCP]Ni(II). The formation of [(pyr)OCP]Ni(II) was accompanied by the C(21)H dehydrogenation step. The nickel(II) ion of [(pyr)OCP]Ni(II), coordinated to a dianionic macrocyclic ligand, is bound by three pyrrolic nitrogens and a trigonally hybridized C(21) atom of the inverted furan. The pyrrole-appended O-confused carbaporphyrin acts as a monoanionic ligand toward zinc(II) and cadmium(II) cations. Three nitrogen atoms and the C(21)H fragment of the inverted furan occupy equatorial positions. In (1)H NMR spectra, the unique inner C(21)H resonances of the inverted furan ring are located at 0.15 ppm for [(pyr)OCPH]Zn(II)Cl, and at 0.21 ppm for [(pyr)OCPH]Cd(II)Cl. The proximity of the furan fragment to the metal ion induces direct scalar couplings between the spin-active nucleus of the metal ((111/113)Cd) and the adjacent (1)H nucleus. The interaction of the metal ion and C(21)H was also reflected by significant changes in carbon chemical shifts ([(pyr)OCPH]Zn(II)Cl, 78.3 ppm; [(pyr)OCPH]Cd(II)Cl, 81.4 ppm; the free base, 101.3 ppm). The density functional theory (DFT) has been applied to model the molecular structures of zinc(II) and cadmium(II) complexes of O-confused oxaporphyrin with an appended pyrrole ring. The Cd...C(21) distance in the optimized structure exceeds the typical Cd-C bond lengths, but is much shorter than the corresponding van der Waals contact. PMID- 16363848 TI - A Mets motif peptide found in copper transport proteins selectively binds Cu(I) with methionine-only coordination. AB - Mets motifs, which refer to methionine-rich sequences found in the high-affinity copper transporter Ctr1, also appear in other proteins involved in copper trafficking and homeostasis, including other Ctrs as well as Pco and Cop proteins isolated from copper-resistant bacteria. To understand the coordination chemistry utilized by these proteins, we studied the copper binding properties of a peptide labeled Mets7-PcoC with the sequence Met-Thr-Gly-Met-Lys-Gly-Met-Ser. By comparing this sequence to a series of mutants containing noncoordinating norleucine in place of methionine, we confirm that all three methionine residues are involved in a thioether-only binding site that is selective for Cu(I). Two independent methods, one based on mass spectrometry and one based on rate differences for the copper-catalyzed oxidation of ascorbic acid, provide an effective K(D) of approximately 2.5 microM at pH 4.5 for the 1:1 complex of Mets7 PcoC with Cu(I). These results establish that a relatively simple peptide containing an MX(2)MX(2)M motif is sufficient to bind Cu(I) with an affinity that corresponds well with its proposed biological function of extracellular copper acquisition. PMID- 16363849 TI - Magnetism, electron paramagnetic resonance, electrochemistry, and mass spectrometry of the pentacopper(II)-substituted tungstosilicate [Cu5(OH)4(H2O)2(A alpha-SiW9O33)2]10-, a model five-spin frustrated cluster. AB - The dimeric, pentacopper(II)-substituted tungstosilicate [Cu(5)(OH)(4)(H(2)O)(2)(A-alpha-SiW(9)O(33))(2)](10)(-) (1) has been characterized by single-crystal X-ray diffraction, elemental analysis, IR, electrochemistry, magnetic measurements, electron paramagnetic resonance (EPR), and mass spectrometry (MS). Magnetization and high-field EPR measurements reveal that the pentameric copper core {Cu(5)(OH)(4)(H(2)O)(2)}(6+) of 1 exhibits strong antiferromagnetic interactions (J(a) = -51 +/- 6 cm(-)(1), J(b) = -104 +/- 1 cm( )(1), and J(c) = -55 +/- 3 cm(-)(1)) resulting in a spin S(T) = (1)/(2) ground state. EPR data show that the unpaired electron spin density is localized on the spin-frustrated apical Cu(2+) ion with g(zz) = 2.4073 +/- 0.0005, g(yy) = 2.0672 +/- 0.0005, g(xx) = 2.0240 +/- 0.0005, and A(zz) = -340 +/- 20 MHz (-0.0113 cm( )(1)). 1 can therefore be considered as a model system for a five-spin, electronically coupled, spin-frustrated system. Polyanion 1, which is stable over a wide pH domain (pH 1-7), was characterized by cyclic voltammetry (CV) in a pH 5 medium. Its CV was constituted by an initial two-step reduction of the Cu(2+) centers to Cu(0) through Cu(+), followed at more negative potential by the redox processes of the W centers. Controlled potential coulometry of 1 allows for the reduction of the five Cu(2+) centers, as seen by consumption of 10.05 +/- 0.05 electrons per molecule. Polyanion 1 triggers efficiently the electrocatalytic reduction of nitrate and nitrite, and it also catalyzes the reduction of N(2)O. To our knowledge, this is the first example of N(2)O catalytic reduction by a polyoxoanion. Fourier transform ion cyclotron resonance MS was used to unambiguously assign the molecular weight of the solution-phase species 1 and the oxidation states of the Cu atoms in the central {Cu(5)(OH)(4)(H(2)O)(2)}(6+) core. Infrared (IR) multiphoton dissociation MS/MS of 1 showed evidence of a condensation process similar to bronze formation at low irradiation intensity. Higher IR intensity resulted in the formation of stable fragments consistent with those previously observed in the solution chemistry of polyoxoanions. PMID- 16363850 TI - Acetylene cyclotrimerization by early second-row transition metals in the gas phase. A theoretical study. AB - The acetylene cyclotrimerization reaction mediated by the left-hand-side bare transition metal atoms Y, Zr, Nb, and Mo has been studied theoretically, employing DFT in its B3LYP formulation. The complete reaction mechanism has been analyzed, identifying intermediates and transition states. Both the ground spin state and at least one low-lying excited state have been considered to establish whether possible spin crossings between surfaces of different multiplicity can occur. Our results show that the overall reaction is highly favorable from a thermodynamic point of view and ground state transition states lie always below the energy limit represented by ground state reactants. After the activation of two acetylene molecules and formation of a bis-ligated complex, the reaction proceeds to give a metallacycle intermediate, as the alternative formation of a cyclobutadiene complex is energetically disfavored. All the examined reaction paths involve formation of a metallacycloheptatriene intermediate that in turn generates a metal-benzene adduct from which finally benzene is released. Similarities and differences in the behaviors of the considered four metal atoms have been examined. PMID- 16363851 TI - Structure-controlled solventless thermolytic synthesis of uniform silver nanodisks. AB - Monodisperse silver nanodisks are synthesized on the gram scale from a well characterized layered silver thiolate precursor via thermolysis at 180-225 degrees C under a N(2) atmosphere. XRD, TEM, HRTEM, and AFM analyses indicate that the nanodisks generated at 180 degrees C over 2 h have an average diameter of about 16.1 nm (sigma = +/-12%) and a thickness of 2.3 nm (sigma = +/-14%), and they lie on their (111) faces. The disk shape is considered to be predestined by the crystal structure of the precursor. Important aspects regarding the stability of the precursor, the thermolysis temperature, and the annealing time, as well as a possible conversion mechanism, are discussed. PMID- 16363852 TI - A kinetically controlled trans bifunctionalized organoimido derivative of the Lindqvist-type hexamolybdate: synthesis, spectroscopic characterization, and crystal structure of (n-Bu4N)2{trans-[Mo6O17(NAr)2]} (Ar = 2,6-dimethylphenyl). AB - A kinetically controlled trans bifunctionalized organoimido derivative of hexamolybdate, (n-Bu(4)N)(2){trans-[Mo(6)O(17)(NAr)(2)]} (Ar = 2,6 dimethylphenyl) 1, in which the two 2,6-dimethylphenylimido groups are bonded to hexamolybdate at the trans positions, has been successfully synthesized in ca. 60% yield under mild reaction conditions. Its trans structure has been confirmed by a single-crystal X-ray diffraction study. In the crystals, cluster anions of 1 self-assemble into a 3D netlike structure via two different kinds of C-H...O hydrogen bondings, in which 1D supramolecular rectangular channels containing tetrabutylammonium cations form along the a axis. Compound 1 has also been characterized by (1)H NMR, IR, and UV-vis spectroscopic studies. UV-vis-near-IR reflectance spectroscopy measurements reveal the compound's nature of semiconductivity with an optical energy gap of 2.55 eV. PMID- 16363853 TI - Synthesis, characterization, and luminescence properties of a new series of Eu3+ containing macrocycles. AB - The synthesis and structural characterization of a series of neutral Eu(3+) containing macrocyclic complexes, Eu(4)-Eu(7), are reported. The synthetic pathway herein allows for the size and functionality of the macrocycle to be tailored in one step from a common precursor (N,N' '-bis(p isothiocyanatobenzylcarbamoylmethyl)diethylenetriamine-N,N'N' '-triacetic acid, (3) in high yield. The macrocyclic ligands 4-7 have within their structure a bis amide derivative of diethylenetriaminepentaacetic acid (DTPA) functioning as the europium chelate that is bridged through thiourea groups by either a butyl (4), hexyl (5), octyl (6), or m-benzyl (7) linker. The two thiourea groups were designed into the host macrocycle to serve as hydrogen-bond donors to potential guest molecules that may alter the luminescence properties of the parent macrocycle. Characterization of the luminescence of Eu(4)-Eu(7) reveals an antenna effect from the ligand, and the luminescence lifetime data reveals the presence of one coordinated water molecule in aqueous solution. PMID- 16363854 TI - Synthesis, structures, and magnetic properties of the copper(II), cobalt(II), and manganese(II) complexes with 9-acridinecarboxylate and 4-quinolinecarboxylate ligands. AB - To explore the relationships between the structures of ligands and their complexes, we have synthesized and characterized a series of metal complexes with two structurally related ligands, 9-acridinecarboxylic acid (HL(1)) and 4 quinolinecarboxylate acid (HL(2)), [Cu(2)(mu(2)-OMe)(2)(L(1))(2)(H(2)O)(0.69)](n) 1, [Cu(2)(L(1))(4)(CH(3)OH)(2)] 2, [Cu(3)(L(1))(6)(CH(3)OH)(6)].3H(2)O 3, [Mn(3)(L(1))(6)(CH(3)OH)(6)].3H(2)O 4, [Co(3)(L(1))(6)(CH(3)OH)(6)].3H(2)O 5, [Cu(L(2))(2)](n) 6, [Mn(L(2))(2)(H(2)O)](n) 7, and [Co(L(2))(2)(H(2)O)](n) 8. 1 is a three-dimensional (3D) polymer with an interpenetrating NbO type network showing one-dimensional (1D) channels, whereas 2 and 3 take bi- and trinuclear structures, respectively, because of the differences in basicity of the reaction systems in preparing the three complexes. 4 and 5 have trinuclear structures similar to that of 3. In 1-5, ligand L(1) performs different coordination modes with N,O-bridging in 1 and O,O'-bridging in 2-5, and the metal ions also show different coordination geometries: square planar in 1, square pyramidal in 2, and octahedral in 3-5. 6 has a two-dimensional structure containing (4,4) grids in which L(2) adopts the N,O-bridging mode and the Cu(II) center takes square planar geometry. 7 and 8 are isostructural complexes showing 1D chain structures, with L(2) adopting the O,O-bridging mode. In addition, the intermolecular O-H...N hydrogen bonds and pi-pi stacking interactions further extend the complexes (except 1 and 6), forming 3D structures. The magnetic properties of 2-7 have been investigated and discussed in detail. PMID- 16363855 TI - Electrochemical and spectroscopic studies of the chloro and oxochloro complex formation of Nb(V) and Ta(V) in NaCl-AlCl3 melts. AB - The equilibrium constant for the chloro complex formation of Nb(V) NbCl6-<-- >NbCl5+Cl- (i) in NaCl-AlCl3 melts at 175 degrees C was found to be pKi = 2.86(5). The oxochloro complex formation of Nb(V) and Ta(V) in NaCl-AlCl3 melts at 175 degrees C could be explained by the following equilibria: MOCl4- <- >MOCl3+Cl- (ii) MOCl3<-->MOCl2(+)+Cl- (iii) where M = Nb and Ta. The equilibrium constants determined by potentiometric measurements with chlorine-chloride electrodes were, for M = Nb, pKii = 2.21(4) and pKiii = 3.95(5) and, for M = Ta, pKii = 2.743(15) and pKiii = 4.521(13). NbCl6- has two bands in the UV-vis region, a strong one at 34.7 x 10(3) cm-1 and a weaker one at 41.6 x 10(3) cm-1. The MOCl4- complexes showed in the case of Nb(V) absorption bands at 32.7 and 42.9 x 10(3) cm-1 and in the case of Ta(V) at 38.6 and 48.1 x 10(3) cm-1. PMID- 16363856 TI - Tetra-2,3-pyrazinoporphyrazines with externally appended pyridine rings. 3. A new highly electron-deficient octacationic macrocycle: tetrakis-2,3-[5,6-di{2-(N methyl)pyridiniumyl}pyrazino]porphyrazine, [(2-Mepy)8TPyzPzH(2)]8+. AB - A new octacationic macrocycle, tetrakis-2,3-[5,6-di{2-(N methyl)pyridiniumyl}pyrazino]porphyrazine, was obtained in its hydrated form as the water-soluble iodide salt. This compound, abbreviated as [(2 Mepy)(8)TPyzPzH(2)](I(8)).8H(2)O (2-Mepy = 2(N-methyl)pyridiniumyl moiety), was obtained by demetalation of the corresponding Mg(II) complex, [(2 Mepy)(8)TPyzPzMg(H(2)O)](I(8)).5H(2)O, which in turn was prepared from its corresponding neutral hydrated species tetrakis-2,3-[5,6-di(2 pyridyl)pyrazino]porphyrazinato(monoaquo)magnesium(II), [Py(8)TPyzPzMg(H(2)O)].4H(2)O, by reaction with CH(3)I in N,N-dimethylformamide. The quaternization reactions by using CH(3)I or methyl p-toluenesulfonate were also conducted on the monomeric precursor 2,3-dicyano-5,6-di(2-pyridyl)-1,4 pyrazine, [(CN)(2)Py(2)Pyz], with formation of the monoquaternized ion [(CN)(2)Py(2-Mepy)Pyz](+) neutralized by iodide and p-toluenesulfonate anions. Single-crystal X-ray work allowed elucidation of the structure of the two salt like species. The diquaternized ion [(CN)(2)(2-Mepy)(2)Pyz](2+) could also be obtained as a p-toluenesulfonate salt, but attempts at direct macrocyclization of this dicationic species were unsuccessful. The iodide salt [(2 Mepy)(8)TPyzPzH(2)](I(8)).8H(2)O is water-soluble, with different solubilities depending on the range of pH explored. It was established that the macrocycle [(2 Mepy)(8)TPyzPzH(2)](8+) undergoes facile deprotonation and behaves as a strong acid. Aggregation phenomena are observed for both the octacation [(2 Mepy)(8)TPyzPzH(2)](8+) and its corresponding centrally deprotonated species [(2 Mepy)(8)TPyzPz](6+). Nevertheless, both cationic moieties exist in their monomeric form under specific experimental conditions. UV-visible monitored titrations with NaOH provide information about the type of protonation/deprotonation equilibria which are complicated by the occurrence of aggregation phenomena. PMID- 16363857 TI - Tetra-2,3-pyrazinoporphyrazines with externally appended pyridine rings. 4. UV visible spectral and electrochemical evidence of the remarkable electron deficient properties of the new tetrakis-2,3-[5,6-di{2-(N methyl)pyridiniumyl}pyrazino]porphyrazinatometal octacations, [(2 Mepy)8TPyzPzM]8+ (M = MgII(H2O), CoII, CuII, ZnII). AB - Metal derivatives of the octacationic tetrakis-2,3-[5,6-di{2-(N methyl)pyridiniumyl}pyrazino]porphyrazine macrocycle [(2-Mepy)(8)TPyzPzH(2)](8+) (2-Mepy = 2-(N-methyl)pyridiniumyl ring) isolated as water-soluble hydrated iodide salts of the general formula [(2-Mepy)(8)TPyzPzM](I(8)).xH(2)O, (M = Mg(II)(H(2)O), Co(II), Cu(II), Zn(II); x = 2-5) were prepared from the corresponding neutral complexes [Py(8)TPyzPzM].xH(2)O previously reported. Reaction of these complexes with CH(3)I in N,N-dimethylformamide under mild conditions led to full quaternization of all eight pyridine N atoms and formation of the octacations [(2-Mepy)(8)TPyzPzM](8+). Clathrated water molecules could be eliminated from the species [(2-Mepy)(8)TPyzPzM](I(8)).xH(2)O by mild heating ( Co(I) process, but the site of electron transfer is reversed and the final product upon a further one-electron reduction is formulated as a Co(II) dianion as opposed to a Co(I) pi-anion radical. This sequence is similar to what was earlier reported for reduction of the same compound in pyridine. Reversible one-electron oxidations are also observed for the unmethylated species [Py(8)TPyzPzM].xH(2)O where M = Co(II) and Mn(II) in DMSO. Remarkably, the octacationic macrocycles [(2 Mepy)(8)TPyzPzM](I(8)).xH(2)O, (M = Mg(II)(H(2)O), Co(II), Cu(II), and Zn(II); x = 2-5) are more easily reduced at any step of the reduction than the corresponding unquaternized species with the same metal ion. This indicates a higher tendency to stepwise electron uptake after the quaternization process, which enhances the charge redistribution capability within the species formed by the electroreduction. PMID- 16363858 TI - Stereochemical investigations of a novel class of chiral phosphapalladacycle complexes derived from 1-[(2,5-dimethyl)phenyl]ethyldiphenylphosphine. AB - The phosphapalladacycle derived from 1-(2',5' dimethylphenyl)ethyldiphenylphosphine has been prepared in the optically active and racemic forms. The phosphine was synthesized as a racemate by the treatment of 1-chloro-1-(2',5'-dimethylphenyl)ethane with sodium diphenylphosphide in THF. The racemic phosphapalladacycle was subsequently obtained as the chloro-bridged dimer by the treatment of the phosphine with palladium(II) acetate followed by anion metathesis with lithium chloride. Alternatively, the phosphine could be optically resolved via metal complexation using (R,R)-bis(mu-chloro)bis{1-[1-(N,N dimethylamino)ethyl]naphthyl-C(2),N}dipalladium(II) as the resolving agent. An efficient separation of the resulting diastereomeric complexes was achieved by silica gel chromatography. The obtained optically resolved diastereomers were next subject to chemoselective removal of the (R)-N,N-(dimethylamino)-1-(1 naphthyl)ethylaminate auxiliary by treatment with concentrated hydrochloric acid. This process yielded the binuclear dimer complexes containing the resolved eta(1) P ligand. Cyclopalladation of the coordinated phosphine could next be performed by treatment of its eta(1)-P binuclear dimer with silver(I) hexafluorophosphate(V) in a dichloromethane/water mixture followed by treatment with lithium chloride, giving rise to a pair of optically pure enantiomeric dimers with [alpha](D) -322 and +319 degrees in CH(2)Cl(2). Despite the possibilities of the phosphine to attain a five- membered structure by ortho palladation or a six-membered ring formation by aliphatic C-H bond activation, only the former was observed. X-ray crystallographic data of the meso dimer and an acetylacetonate derivative indicated that the phosphapalladacycle alpha-C methyl substituent was axially located. The 2-D (1)H-(1)H ROESY spectrum of the acetylacetonate derivative further revealed that the phosphapalladacycle was conformationally rigid in CDCl(3). PMID- 16363859 TI - Dithiolate complexes of manganese and rhenium: X-ray structure and properties of an unusual mixed valence cluster Mn3(CO)6(mu-eta2-SCH2CH2CH2S)3. AB - Treatment of Mn(2)(CO)(10) with 3,4-toluenedithiol and 1,2-ethanedithiol in the presence of Me(3)NO.2H(2)O in CH(2)Cl(2) at room temperature afforded the dinuclear complexes Mn(2)(CO)(6)(mu-eta(4)-SC(6)H(3)(CH(3))S-SC(6)H(3)(CH(3))S) (1), and Mn(2)(CO)(6)(mu-eta(4)-SCH(2)CH(2)S-SCH(2)CH(2)S) (2), respectively. Similar reactions of Re(2)(CO)(10) with 3,4-toluenedithiol, 1,2-benzenedithiol, and 1,2-ethanedithiol yielded the dirhenium complexes Re(2)(CO)(6)(mu-eta(4) SC(6)H(3)(CH(3))S-SC(6)H(3)(CH(3))S) (3), Re(2)(CO)(6)(mu-eta(4)-SC(6)H(4)S SC(6)H(4)S) (4), and Re(2)(CO)(6)(SCH(2)CH(2)S-SCH(2)CH(2)S) (5), respectively. In contrast, treatment of Mn(2)(CO)(10) with 1,3-propanedithiol afforded the trimanganese compound Mn(3)(CO)(6)(mu-eta(2)-SCH(2)CH(2)CH(2)S)(3) (6), whereas Re(2)(CO)(10) gave only intractable materials. The molecular structures of 1, 3, and 6 have been determined by single-crystal X-ray diffraction studies. The dimanganese and dirhenium carbonyl compounds 1-5contain a binucleating disulfide ligand, formed by interligand disulfide bond formation between two dithiolate ligands identical in structure to that of the previously reported dimanganese complex Mn(2)(CO)(6)(mu-eta(4)-SC(6)H(4)S-SC(6)H(4)S). Complex 6, on the other hand, forms a unique example of a mixed-valence trimangenese carbonyl compound containing three bridging 1,3-propanedithiolate ligands. The solution properties of 6 have been investigated by UV-vis and EPR spectroscopies as well as electrochemical techniques. PMID- 16363860 TI - Synthesis and growth mechanisms of one-dimensional strontium hydroxyapatite nanostructures. AB - Strontium hydroxyapatite (SrHAp) nanowires with an aspect ratio of several hundreds were synthesized by controlling the growth conditions during a hydrothermal process. In the strontium phosphate system, it was found that the phase evolution changed with pH and that the aspect ratio of SrHAp was affected by the phases present before heating. Since the conditions for SrHAp nucleation prohibits one-dimensional growth, it was impossible to grow large-scale SrHAp nanowires using routine hydrothermal methods. Through thermodynamic considerations, the mechanisms of nanowire formation appear to involve the rapid release of the stored chemical potential in a metastable phase, which promotes the anisotropic growth of the most stable SrHAp nanostructures. Thereby, the conditions for both the nucleation of the SrHAp phase and the anisotropic growth were determined simultaneously, and considerable quantities of SrHAp nanowires were synthesized. PMID- 16363861 TI - X-ray structures and homolysis of some alkylcobalt(III) phthalocyanine complexes. AB - The first crystallographic data for sigma-bonded alkylcobalt(III) phthalocyanine complexes are reported. A single-crystal X-ray structure of CH(3)CH(2)Co(III)Pc (Pc = dianion of phthalocyanine) reveals that the solid consists of centrosymmetric face-to-face dimers in which the CH(3)CH(2)Co(III)Pc units retain their square pyramidal geometry. The structure appears to be the first one reported for a five-coordinate RCo(III)(chelate) complex with an electron deficient equatorial system. The Co-C bond in CH(3)CH(2)Co(III)Pc (2.031(5) A) is the longest found in five-coordinate RCo(III)(chel) complexes (R = simple primary alkyl group). Another X-ray study demonstrates that CH(3)Co(III)Pc(py) has a distorted octahedral geometry with axial bonds of very similar length to those in methylcobalamin. The axial bonds are shorter than those in its octaethylporphyrin analogue, in accordance with a weaker trans axial influence in six-coordinate complexes containing an electron-deficient phthalocyanine equatorial ligand. A different trend has been observed for five-coordinate RCo(III)(chel) complexes: electron-rich equatorial systems seem to make the Co-C axial bond shorter. Kinetic data for the homolysis of RCo(III)Pc complexes (R = Me, Et) in dimethylacetamide are also reported. Homolysis of ethyl derivatives is faster. The Co-C bond dissociation energies (BDEs) for the pyridine adducts of the methyl and the ethyl derivative are 30 +/- 1 and 29 +/- 1 kcal/mol, respectively. The BDE for CH(3)CoPc(py) is considerably lower than that for MeCbl despite the very similar lengths of the axial bonds in the two complexes. The results of this work do not support any correlation between the Co-C bond length and the bond strength as defined by BDE. PMID- 16363862 TI - Substituent effects on indium-phosphorus bonding in (4-RC6H4S)3In.PR'3 adducts (R = H, Me, F; R' = Et, Cy, Ph): a spectroscopic, structural, and thermal decomposition study. AB - The tris(arylthiolate)indium(III) complexes (4-RC(6)H(4)S)(3)In [R = H (5), Me (6), F (7)] were prepared from the 2:3 reaction of elemental indium and the corresponding aryl disulfide in methanol. Reaction of 5-7 with 2 equiv of the appropriate triorganylphosphine in benzene or toluene resulted in isolation of the indium-phosphine adduct series (4-RC(6)H(4)S)(3)In.PR'(3) [R = H, R' = Et (5a), Cy (5b), Ph (5c); R = Me, R' = Et (6a), Cy (6b), Ph (6c); R = F, R' = Et (7a), Cy (7b), Ph (7c)]. These compounds were characterized via elemental analysis, FT-IR, FT-Raman, solution (1)H, (13)C{(1)H}, (31)P{(1)H}, and (19)F (7a c) NMR spectroscopy, and X-ray crystallography (5c, 6a, 6c, and 7a). NMR spectra show retention of the In-P bond in benzene-d(6) solution, with phosphine (31)P{(1)H} signals shifted downfield compared to the uncoordinated ligand. The X ray structures show monomeric 1:1 adduct complexes in all cases. The In-P bond distance [2.5863(5)-2.6493(12) A] is influenced significantly by the phosphine substituents but is unaffected by the substituted phenylthiolate ligand. Relatively low melting points (88-130 degrees C) are observed for all adducts, while high-temperature thermal decomposition is observed for the indium thiolate reactants 5-7. DSC/TGA and EI-MS data show a two-step thermal decomposition process, involving an initial loss of the phosphine moiety followed by loss of thiolate ligand. PMID- 16363863 TI - Versatility of dithiophosphates in the syntheses of copper(I) complexes with bis(diphenylphosphino)alkanes: abstraction of chloride from dichloromethane. AB - Reactions of [Cu(CH(3)CN)(4)]X (X = PF(6), BF(4)) with bis(diphenylphosphino)methane (dppm = Ph(2)PCH(2)PPh(2)) and ammonium dialkyldithiophosphates, (NH(4))[S(2)P(OR)(2)] (R = Et, (i)Pr), yield a series of novel Cu(I) polynuclear complexes, trinuclear [Cu(3)(mu-dppm)(3)(mu(3) Cl){S(2)P(OEt)(2)}] (PF(6)) 1 and [Cu(3)(mu-dppm)(2){S(2)P(OR)(2)}(2)](PF(6)) (R = Et, 2; (i)Pr, 3), tetranuclear [Cu(4)(mu-dppm)(2) {S(2)P(OEt)(2)}(4)] 4, and hexanuclear [Cu(6)(mu-dppm)(2)(mu(4)-Cl){S(2)P(O(i)()Pr)(2)}(4)](BF(4)) 5. Similarly, the reaction of [Cu(2)(mu-L-L)(2)(CH(3)CN)(2)](PF(6))(2) (L-L, dppm, dppe = Ph(2)PCH(2)CH(2)PPh(2)) with (NH(4))[S(2)P(OR)(2)] yields dinuclear [Cu(2)(mu-dppm)(2){S(2)P(OR)(2)}(2)] 6 (R= (i)Pr, 6A; Et, 6B), trinuclear [Cu(3)(mu-dppe)(3)(mu-Cl)(2){S(2)P(O(i)Pr)(2)}] 9, and polymeric [Cu(mu(2) dppe){S(2)P(OR)(2)}](n) (R = Et, 7; (i)Pr, 8) complexes. The formation of 1 and 5 involved the abstraction of chloride from dichloromethane when the Cu/S(2)P(OR)(2) ratio exceeded 1, but when ratio was 1:1, no Cl abstraction occurred, as in compound 4. Compound 9, however, was obtained as a 12% byproduct in the synthesis of 8 using a 1:1:1 ratio of Cu/dppe/S(2)P(O(i)Pr)(2). The chloride binds to Cu atoms in a mu(3)-Cl mode by capping one face of the Cu(3) triangle of cluster 1. A mu(4)-Cl caps a single tetragonal face of the trigonal prism of cluster 5, and in the cluster 9, two chlorides bond in mu(2)-Cl modes. Both clusters 2 and 3 exhibit the mu(3)-S mode of bonding for dtp ligands. Only cluster 5 exhibited close Cu...Cu contacts (2.997-3.0238 A). All of compounds were characterized by single-crystal X-ray diffraction and pertinent crystallographic data for 1, 5, and 9 are are follows: (1) C(79)H(76)ClCu(3)F(6)O(2)P(8)S(2), triclinic, P, a = 11.213(1) A, b = 14.142(1) A, c = 25.910(2) A, alpha = 95.328(2) degrees , beta = 99.594(2) degrees , gamma = 102.581(2) degrees , V = 3918.2(6) A(3), Z = 2; (5) C(74)H(100)BClCu(6)F(4)O(8)P(8)S(8), monoclinic, P2(1)/n, a = 25.198(4) A, b = 15.990(3) A, c = 25.421(4) A, beta = 106.027(3) degrees , V = 9845(3)A(3), Z = 4; (9) C(84)H(86)Cl(2)Cu(3)O(2)P(7)S(2), monoclinic, C2/c, with a = 24.965(3) A, b = 17.058(2) A, c = 20.253(2) A, beta = 95.351(4) degrees , V = 8587.4(17)A(3), Z = 4. PMID- 16363864 TI - Novel rhenium chelate system derived from dimercaptosuccinic acid for the selective labeling of biomolecules. AB - This work is part of an effort to develop chelating agents for stable binding and easy conjugation of Re-188 to biologically interesting structures. Starting from the well-known in vivo stability of [(188)ReO(DMSA)(2)](-), we want to exploit this coordination system for the design of (188)ReO(V) chelates, which are stable toward reoxidation to perrhenate and toward ligand exchange under all conditions of radiopharmaceutical development. Therefore, a new type of tetradentate ligand has been synthesized by bridging two molecules of N,N'-diisobutyl-2,3 dimercaptosuccinamide with N-(3-aminopropyl)propane-1,3-diamine. The resulting stereoisomeric tetrathiolato S(4) ligand of composition ((i)()Bu)(2)N(O)C-C(SH) C(SH)-C(O)NH-(CH(2))(3)-NH-(CH(2))(3)-NHC(O)-C(SH)-C(SH)-C(O)N((i)Bu)(2) forms anionic five-coordinate oxorhenium(V) complexes by a ligand-exchange reaction of NBu(4)[ReOCl(4)] in methanol. In the absence of a base, the compounds were isolated as "betaine", [ReO(S(4))], with the protonated nitrogen of the bridge serving as an internal "counterion". Two representatives have been fully characterized in both the solid and solution states and found to adopt the expected square-pyramidal coordination geometry. The equatorial plane is formed by four thiolate sulfur atoms, whereas the oxygen occupies the apical position. The orientation of the metal oxo group is exo in relation to the carbamido groups in both isomers. Both complexes are stereoisomeric regarding the junction of the triamine chain. PMID- 16363865 TI - Synthesis of WO3 nanorods by reacting WO(OMe)4 under autogenic pressure at elevated temperature followed by annealing. AB - This article reports on the fabrication of WO(3) nanorods using an efficient straightforward synthetic technique, without a catalyst, and using a single precursor. The thermal dissociation of WO(OMe)(4) at 700 degrees C in a closed Swagelok cell under an air/inert atmosphere yielded W(18)O(49) nanorods. Annealing of W(18)O(49) at 500 degrees C under an air atmosphere led to the formation of pure WO(3) nanorods. The obtained products are characterized by morphological (scanning electron microscopy and transmission electron microscopy), structural (X-ray diffraction analysis, high-resolution scanning electron microscopy, and Raman spectroscopy), and compositional [energy dispersive X-ray and elemental (C, H, N, S) analysis] measurements. The mechanism of the formation of nonstoichiometric W(18)O(49) nanorods is supported by the measured analytical data and several control experiments. PMID- 16363866 TI - Nitric oxide production by visible light irradiation of aqueous solution of nitrosyl ruthenium complexes. AB - [Ru(II)L(NH(3))(4)(pz)Ru(II)(bpy)(2)(NO)](PF(6))(5) (L is NH(3), py, or 4-acpy) was prepared with good yields in a straightforward way by mixing an equimolar ratio of cis-[Ru(NO(2))(bpy)(2)(NO)](PF(6))(2), sodium azide (NaN(3)), and trans [RuL(NH(3))(4)(pz)] (PF(6))(2) in acetone. These binuclear compounds display nu(NO) at ca. 1945 cm(-)(1), indicating that the nitrosyl group exhibits a sufficiently high degree of nitrosonium ion (NO(+)). The electronic spectrum of the [Ru(II)L(NH(3))(4)(pz)Ru(II)(bpy)(2)(NO)](5+) complex in aqueous solution displays the bands in the ultraviolet and visible regions typical of intraligand and metal-to-ligand charge transfers, respectively. Cyclic voltammograms of the binuclear complexes in acetonitrile give evidence of three one-electron redox processes consisting of one oxidation due to the Ru(2+/3+) redox couple and two reductions concerning the nitrosyl ligand. Flash photolysis of the [Ru(II)L(NH(3))(4)(pz)Ru(II)(bpy)(2)(NO)](5+) complex is capable of releasing nitric oxide (NO) upon irradiation at 355 and 532 nm. NO production was detected and quantified by an amperometric technique with a selective electrode (NOmeter). The irradiation at 532 nm leads to NO release as a consequence of a photoinduced electron transfer. All species exhibit similar photochemical behavior, a feature that makes their study extremely important for their future application in the upgrade of photodynamic therapy in living organisms. PMID- 16363867 TI - Oxidation of thioglycolate by [Os(phen)3]3+: an unusual example of redox-mediated aromatic substitution. AB - The aqueous oxidation of thioglycolic acid (TGA) by [Os(phen)(3)](3+) (phen = 1,10-phenanthroline) is catalyzed by traces of ubiquitous Cu(2+) and inhibited by the product [Os(phen)(3)](2+). In the presence of dipicolinic acid (dipic), which thoroughly masks trace Cu(2+) catalysis, and spin trap PBN, the kinetics under anaerobic conditions have been studied in the pH range 1.82-7.32. The rate law is -d[Os(phen)(3)(3+)]/dt = k[TGA](tot)[Os(phen)(3)(3+)], with k = 2{(k(b)K(a1) + k(c)K(a1)K(i))[H(+)] + k(d)K(a1)K(a2)}/{[H(+)](2) + K(a1)[H(+)] + K(a1)K(a2)}; K(a1) and K(a2) are the successive acid dissociation constants of TGA, and K(i) is the tautomerization constant of two TGA monoanions. k(b) + k(c)K(i) = (5.9 +/- 0.3) x 10(3) M(-)(1) s(-)(1), k(d) = (1.6 +/- 0.1) x 10(9) M(-)(1) s(-)(1) at mu = 0.1 M (NaCF(3)SO(3)) and 25 degrees C. The major products in the absence of spin traps are dithiodiglycolic acid, [Os(phen)(3)](2+), and [Os(phen)(2)(phen tga)](2+), where phen-tga is phenanthroline with a TGA substituent. A mechanism is proposed in which neutral TGA is unreactive, the (minor) thiolate form of the TGA monoanion undergoes one-electron oxidation by [Os(phen)(3)](3+) (k(c)), and the dianion of TGA likewise undergoes one-electron oxidation by [Os(phen)(3)](3+) (k(d)). The Marcus cross relationship provides a good account for the magnitude of k(d) in this and related reactions of TGA. [Os(phen)(2)(phen-tga)](2+) is suggested to arise from a post-rate-limiting step involving attack of the TGA(*) radical on [Os(phen)(3)](3+). PMID- 16363868 TI - Coordination versatility of pyridine-functionalized N-heterocyclic carbenes: a detailed study of the different activation procedures. Characterization of new Rh and Ir compounds and study of their catalytic activity. AB - Three different reaction procedures for the coordination of N-n-butyl-N'-(2 pyridylmethyl)imidazolium salt have produced new N-heterocyclic complexes of Rh and Ir. The direct reaction of the imidazolium salt with [IrCl(cod)](2) provides a NHC-Ir(III)-H complex, while transmetalation from a silver-NHC complex and deprotonation with NEt(3) give new NHC complexes of M(I) and M(III) when reacting with [MCl(cod)](2) or [MCl(coe)(2)](2) (M = Rh, Ir). The crystal structures of the biscarbene Rh(III) and Ir(III) complexes are described. The catalytic properties of the compounds obtained have been tested in the hydrosilylation of acetylenes, the cyclization of acetylenic carboxylic acids, and hydrogen transfer to ketones. PMID- 16363869 TI - Synthesis and characterization of new calcium phenylphosphonates and 4 carboxyphenylphosphonates. AB - Three new calcium phenylphosphonates, CaC(6)H(5)PO(3).2H(2)O, Ca(3)(C(6)H(5)PO(3)H)(2)(C(6)H(5)PO(3))(2).4H(2)O, and CaC(6)H(5)PO(3).H(2)O, and two calcium 4-carboxyphenylphosphonates, Ca(HOOCC(6)H(4)PO(3)H)(2) and Ca(3)(OOCC(6)H(4)PO(3))(2).6H(2)O, were prepared. It was found that CaC(6)H(5)PO(3).2H(2)O transformed into previously known Ca(C(6)H(5)PO(3)H)(2) via Ca(3)(C(6)H(5)PO(3)H)(2)(C(6)H(5)PO(3))(2).4H(2)O in the presence of phenylphosphonic acid, and vice versa, Ca(C(6)H(5)PO(3)H)(2) turned into CaC(6)H(5)PO(3).2H(2)O in a weak basic medium. A similar relationship was found between Ca(HOOCC(6)H(4)PO(3)H)(2) and Ca(3)(OOCC(6)H(4)PO(3))(2).6H(2)O; i.e., Ca(3)(OOCC(6)H(4)PO(3))(2).6H(2)O transformed into Ca(HOOCC(6)H(4)PO(3)H)(2) in the presence of 4-carboxyphenylphosphonic acid. On the contrary, Ca(3)(OOCC(6)H(4)PO(3))(2).6H(2)O is formed from Ca(HOOCC(6)H(4)PO(3)H)(2) in the presence of ammonium as a weak base. The structure of Ca(HOOCC(6)H(4)PO(3)H)(2) was solved from X-ray powder diffraction data by an ab initio method using a FOX program. The compound is monoclinic, space group C2/c (No. 15), a = 49.218(3) A, b = 7.7609(4) A, c = 5.4452(3) A, beta = 128.119(3) degrees , and Z = 4. Its structure is one-dimensional with [Ca(2)(HOOCC(6)H(4)PO(3)H)(4)](infinity) ribbons forming basic building blocks. The ribbons are held together by hydrogen bonds between carboxylic groups. PMID- 16363872 TI - Conformational flexibility of the peptide hormone ghrelin in solution and lipid membrane bound: a molecular dynamics study. AB - Human ghrelin is a peptide hormone of 28 aminoacid residues, in which the Ser3 is modified by an octanoyl group. Ghrelin has a major role in the energy metabolism of the human body stimulating growth hormone release as well as food intake. Here we perform molecular dynamics simulations in explicit water and in a DMPC-lipid bilayer/water system in order to structurally characterize this highly flexible peptide and its lipid binding properties. We find a loop structure with residues Glu17 to Lys 20 in the bending region and a short alpha-helix from residues Pro7 to Glu13. The presence of a lipid membrane does not influence these structural features, but reduces the overall flexibility of the molecule as revealed by reduced root mean square fluctuations of the atom coordinates. The octanoyl-side chain does not insert into the lipid membrane but points into the water phase. The peptide binds to the lipid membrane with its bending region involving residues Arg15, Lys16, Glu17, and Ser18. The implications of these results for the binding pocket of the ghrelin receptor are discussed. PMID- 16363873 TI - Cation-mediated interplay of loops in chaperonin-10. AB - The ubiquitously occurring chaperonins consist of a large tetradecameric Chaperonin-60, forming a cylindrical assembly, and a smaller heptameric Chaperonin-10. For a functional protein folding cycle, Chaperonin-10 caps the cylindrical Chaperonin-60 from one end forming an asymmetric complex. The oligomeric assembly of Chaperonin-10 is known to be highly plastic in nature. In Mycobacterium tuberculosis, the plasticity has been shown to be modulated by reversible binding of divalent cations. Binding of cations confers rigidity to the metal binding loop, and also promotes stability of the oligomeric structure. We have probed the conformational effects of cation binding on the Chaperonin-10 structure through fluorescence studies and molecular dynamics simulations. Fluorescence studies show that cation binding induces reduced exposure and flexibility of the dome loop. The simulations corroborate these results and further indicate a complex landscape of correlated motions between different parts of the molecule. They also show a fascinating interplay between two distantly spaced loops, the metal binding "dome loop" and the GroEL-binding "mobile loop", suggesting an important cation-mediated role in the recognition of Chaperonin-60. In the presence of cations the mobile loop appears poised to dock onto the Chaperonin-60 structure. The divalent metal ions may thus act as key elements in the protein folding cycle, and trigger a conformational switch for molecular recognition. PMID- 16363874 TI - In silico structure-based design of a potent and selective small peptide inhibitor of protein tyrosine phosphatase 1B, a novel therapeutic target for obesity and type 2 diabetes mellitus: a computer modeling approach. AB - Protein Tyrosine Phosphatase 1B (PTP1B) has been shown to be a negative regulator of insulin signaling by dephosphorylating key tyrosine residues within the regulatory domain of the beta-subunit of the insulin receptor. Recent gene knockout studies in mice have shown the mice to have increased insulin sensitivity and improved glucose tolerance. Furthermore, these mice also exhibited a resistance to diet induced obesity. Inhibitors of PTP1B would have the potential of enhancing insulin action by prolonging the phosphorylated state of the insulin receptor. In addition, recent clinical studies have shown that the haplotype ACTTCAG0 of the PTPN1 gene, which encodes PTP1B, is a major risk contributor to type 2 diabetes mellitus (T2DM). Thus, there is compelling evidence that small molecule inhibitors of PTP1B may be effective in treating insulin resistance at an early stage, thereby leading to a prevention strategy for T2DM and obesity. Based on the crystal structure of the complex of PTP1B with a known inhibitor, we have identified a tetrapeptide inhibitor with the sequence WKPD. Docking calculations indicate that this peptide is as potent as the existing inhibitors. Moreover, the peptide is also found to be selective for PTP1B with a greatly reduced potency against other biologically important protein tyrosine phosphatases such as PTP-LAR, Calcineurin, and the highly homologous T Cell Protein Tyrosine Phosphatase (TCPTP). Thus the designed tetrapeptide is a suitable lead compound for the development of new drugs against type 2 diabetes and obesity. PMID- 16363875 TI - Protein structure evaluation using an all-atom energy based empirical scoring function. AB - Arriving at the native conformation of a polypeptide chain characterized by minimum most free energy is a problem of long standing interest in protein structure prediction endeavors. Owing to the computational requirements in developing free energy estimates, scoring functions--energy based or statistical- have received considerable renewed attention in recent years for distinguishing native structures of proteins from non-native like structures. Several cleverly designed decoy sets, CASP (Critical Assessment of Techniques for Protein Structure Prediction) structures and homology based internet accessible three dimensional model builders are now available for validating the scoring functions. We describe here an all-atom energy based empirical scoring function and examine its performance on a wide series of publicly available decoys. Barring two protein sequences where native structure is ranked second and seventh, native is identified as the lowest energy structure in 67 protein sequences from among 61,659 decoys belonging to 12 different decoy sets. We further illustrate a potential application of the scoring function in bracketing native-like structures of two small mixed alpha/beta globular proteins starting from sequence and secondary structural information. The scoring function has been web enabled at www.scfbio-iitd.res.in/utility/proteomics/energy.jsp. PMID- 16363876 TI - The increased flexibility of CDR loops generated in antibodies by Congo red complexation favors antigen binding. AB - The dye Congo red and related self-assembling compounds were found to stabilize immune complexes by binding to antibodies currently engaged in complexation to antigen. In our simulations, it was shown that the site that becomes accessible for binding the supramolecular dye ligand is located in the V domain, and is normally occupied by the N-terminal polypeptide chain fragment. The binding of the ligand disrupts the beta-structure in the domain, increasing the plasticity of the antigen-binding site. The higher fluctuation of CDR-bearing loops enhances antigen binding, and allows even low-affinity antibodies to be engaged in immune complexes. Experimental observations of the enhancement effect were supported by theoretical studies using L lambda chain (4BJL-PDB identification) and the L chain from the complex of IgM-rheumatoid factor bound to the CH3 domain of the Fc fragment (1ADQ-PDB identification) as the initial structures for theoretical studies of dye-induced changes. Commercial IgM-type rheumatoid factor (human) and sheep red blood cells with coupled IgG (human) were used for experimental tests aimed to reveal the dye-enhancement effect in this system. The specificity of antigen-antibody interaction enhanced by dye binding was studied using rabbit anti-sheep red cell antibodies to agglutinate red cells of different species. Red blood cells of hoofed mammals (horse, goat) showed weak enhancement of agglutination in the presence of Congo red. Neither agglutination nor enhancement were observed in the case of human red cells. The dye-enhancement capability in the SRBC-antiSRBC system was lost after pepsin-digestion of antibodies producing (Fab)2 fragments still agglutinating red cells. Monoclonal (myeloma) IgG, L lambda chain and ovoalbumin failed to agglutinate red cells, as expected, and showed no enhancement effect. This indicates that the enhancement effect is specific. PMID- 16363877 TI - Structural differentiation of the HIV-1 polyA signals. AB - The Human Immunodeficiency Virus Type 1 (HIV-1) encodes the polyadenylation (polyA) signal (AAUAAA) within the highly conserved untranslated region (UTR) at both 5' and 3' terminals of the viral transcript. In polyadenylation, an RNA transcript is cleaved and then elongated with adenine nucleotides while repression of the 5' signal and utilization of the 3' signal occurs. Because experimental studies have yet to analyze the structures of both 5' and 3' signals from a global perspective, other structural conformations involving these signals may exist and could be pivotal to understanding key functional processes. To distinguish the differential regulation of the 5' and 3' polyA signals, we studied the structural tendencies of both the 5' and 3' UTR in HIV-1. Through computational folding predictions of multiple HIV-1 strains using the Massively Parallel Genetic Algorithm (MPGAfold) capable of dynamically elucidating key alternative conformations, the 5' polyA signal was found to be dominantly occluded in a hairpin loop while the 3' polyA signal showed variability between hairpin and linear conformations with a propensity for the linear structure with an asymmetric internal loop. Furthermore, the energies and predictions of these structures indicate that the polyA signals have some metastable characteristics indicating an ability to switch into different conformations that can regulate viral function. PMID- 16363878 TI - Evidence for long poly(dA).poly(dT) tracts in D. discoideum DNA at high frequencies and their preferential avoidance of nucleosomal DNA core regions. AB - The eukaryote, Dictyostelium discoideum, has one of the most (A+T) rich genomes studied to date. Isolated nuclear D. discoideum DNA (AX3 strain) was used to qualitatively determine the frequency and length distribution of long (dA).(dT) homopolymer tracts in this genome, in comparison to the less (A+T) rich calf thymus and Schistosoma mansoni DNAs that had few observable long tracts. These experimental data accurately reflect the significantly elevated frequencies of long tracts found computationally within the D. discoideum intron and flanking sequences, but not exons. PCR amplification of long (dA).(dT) homopolymer tract containing sequences was carried out. Then experimental biotinylated (dT)18 probe hybridization to the PCR amplified DNA showed that the long (dA).(dT) homopolymer tracts were enriched in D. discoideum sequences only hundreds of base pair in length, under conditions where no equivalent hybridization was observed to S. mansoni DNA or calf DNA sequences. Similar probe hybridization to DNA isolated following micrococcal nuclease digestion of D. discoideum chromatin demonstrated that long (dA).(dT) homopolymer tracts were more highly enriched in nucleosomal DNA lengths that included the internucleosomal linker as compared to shorter linker free mononucleosomal lengths. This observation is in agreement with the frequency of tract spacing results calculated from GenBank sequence data. These frequency data indicate that adjacent long tracts plus the intervening spacer DNA are found at peak lengths (average 42 bp), exactly characteristic of the internucleosomal spacer region of D. discoideum chromatin and are in sufficient number to be found in nearly half of all nucleosomes. Compared to shuffled tract sequence controls, these lengths of adjacent long tracts plus the intervening spacer DNA were found to be significantly enriched. Lesser enrichments are observed at lengths corresponding to adjacent tracts being separated by nucleosomal core length DNA sequences (145-185 bp). These data strongly suggest that adjacent long tracts occur spaced at selected lengths so as to avoid the central core regions of nucleosomes and instead are found localized within internucleosomal DNA linker and core edge regions in D. discoideum chromatin. PMID- 16363879 TI - Data mining of molecular dynamics trajectories of nucleic acids. AB - Analysis, storage, and transfer of molecular dynamic trajectories are becoming the bottleneck of computer simulations. In this paper we discuss different approaches for data mining and data processing of huge trajectory files generated from molecular dynamic simulations of nucleic acids. PMID- 16363880 TI - 'Genomemark': detecting word periodicity in biological sequences. AB - Identifying and predicting the structural characteristics of novel repeats throughout the genome can lend insight into biological function. Specific repeats are believed to have biological significance as a function of their distribution patterns. We have developed 'GenomeMark,' a computer program that detects and statistically analyzes candidate repeats. Specifically, 'GenomeMark' identifies the periodic distribution of unique words, calculating their chi2 and Z-score values. Using 'GenomeMark,' we identified novel sequence words present in tandem throughout genomes. We found that these sequences have remarkable spacer sequence distributions and many were genome specific, validating the genome signature theory. Further analysis confirmed that many of these sequences have a specific biological function. The program is available from the authors upon request and is freely available for non-commercial and academic entities. PMID- 16363881 TI - Acid titrations of poly(dG-dC).poly(dG-dC) in aqueous solution and in a w/o microemulsion. AB - The model polynucleotide poly(dG-dC).poly(dG-dC) (polyGC) was titrated with a strong acid (HCl) in aqueous unbuffered solutions and in the quaternary w/o microemulsion CTAB/n-pentanol/n-hexane/water. The titrations, performed at several concentrations of NaCl in the range 0.005 to 0.600 M, were followed by recording the modifications of the electronic absorption and of the CD spectra (210< or = lambda < or =350 nm) upon addition of the acid. In solution, the polynucleotide undergoes two acid-induced transitions, neither of which corresponds to denaturation of the duplex to single coil. The first transition leads to the Hoogsteen type synG.C+ duplex, while the second leads to the C+.C duplex. The initial B-form of polyGC was recovered by back-titration with NaOH. The apparent pKa values were obtained for both steps of the titration, at all salt concentrations. A reasonably linear dependence of pKa1 and pKa2 from p[NaCl] was obtained, with both pKa values decreasing with increasing ionic strength. In microemulsion, at salt concentrations < or = 0.300 M, an acid-induced transition was observed, matching the first conformational transition recorded also in solution. However, further addition of acid led to denaturation of the protonated duplex. Renaturation of polyGC was obtained by back-titration with NaOH. At salt concentrations > 0.300 M, polyGC is present as a mixture of B-form and psi- aggregates, that slowly separate from the microemulsion. The acid titration induces at first a conformational transition similar to the one observed at low salt or in solution, then denaturation occurs, which is however preceded by the appearance of a transient conformation, that has been tentatively classified as a left-handed Z double helix. PMID- 16363882 TI - Kinetics of ligand binding to nucleic acids. AB - Ligand binding to nucleic acids (NA) is considered as a stationary Markov process. It is shown that the probabilistic description of ligand-NA binding allows one to describe not only the kinetics of the change of number of bound ligands at arbitrary fillings but also to calculate stationary values of the number of bound ligands and its dispersion. The general analysis of absorption isotherms and kinetics of ligand binding to NA make it possible to determine of rate constants of ligand-NA complex formation and dissociation. PMID- 16363884 TI - Therapy of chronic lymphocytic leukaemia with purine nucleoside analogues: facts and controversies. AB - Chronic lymphocytic leukaemia (CLL) is a neoplastic disease of unknown aetiology characterised by an absolute lymphocytosis in peripheral blood and bone marrow. The disease is diagnosed most commonly in the elderly with the median age at diagnosis being about 65 years. The purine nucleoside analogues (PNAs) fludarabine, cladribine (2-chlorodeoxyadenosine) and pentostatin (2' deoxycoformycin) are highly active in CLL, both in previously treated and in refractory or relapsed patients. These three agents share similar chemical structures and mechanisms of action such as induction of apoptosis. However, they also exhibit significant differences, especially in their interactions with enzymes involved in adenosine and deoxyadenosine metabolism. Recent randomised studies suggest that fludarabine and cladribine have similar activity in CLL. However, clinical observations indicate the existence of cross-resistance between fludarabine and cladribine. Patients who received PNAs as their initial therapy and achieved long-lasting response can be successfully retreated with the same agent. PNAs administered in combination with other chemotherapeutic agents and/or monoclonal antibodies may produce higher response rates, including complete response (CR) or molecular CR, compared with PNAs alone or other treatment regimens. Management decisions are more difficult in elderly patients because of the apparent increase in toxicity of PNAs in this population. In elderly patients, we recommend chlorambucil as the first-line treatment, with PNAs in lower doses in refractory or relapsed patients. Myelosuppression and infections, including opportunistic varieties, are the most frequent adverse effects in patients with CLL treated with PNAs. Therefore, some investigators recommend routine antibacterial and antiviral prophylaxis during and after PNA treatment. This review presents current results and treatment strategies with the use of PNAs in CLL, especially in elderly patients. PMID- 16363885 TI - Overactive bladder in the elderly: a guide to pharmacological management. AB - Overactive bladder (OAB) is a common condition characterised by the symptoms of urinary frequency and urgency, with or without urge incontinence and nocturia. The prevalence of OAB increases markedly with age in both men and women. OAB can have a detrimental effect on physical functioning and psychological well-being, as well as significantly reducing quality of life. Antimuscarinic therapy -- with or without behavioural therapy -- represents the most common treatment for patients with OAB. Several antimuscarinic agents are currently available for the treatment of OAB in adults, including oxybutynin, tolterodine, trospium chloride, darifenacin and solifenacin. The antimuscarinics all appear to exert their clinical effect through inhibition of the bladder muscarinic receptors, but they vary both in structure and in their functional profile. While efficacy has been demonstrated in adult populations (including patients >65 years of age), few studies have been reported specifically in a geriatric population, and antimuscarinics are often underutilised in the elderly despite the marked increase in the prevalence of OAB in this age group. One explanation for this apparent underuse of an effective treatment option may be concerns about the frequency of anticholinergic adverse events, such as dry mouth; the likelihood of detrimental CNS effects, including cognitive impairment and sleep disturbances; and the potential for harmful interactions with existing pharmacotherapy. When selecting an antimuscarinic agent for the management of an elderly patient presenting with OAB, in addition to considering evidence of clinical efficacy and tolerability, issues of safety specific to an older population should be borne in mind. In particular, the likelihood of detrimental CNS effects should be considered, including cognitive impairment and sleep disturbances, secondary to anticholinergic load. Oxybutynin and tolterodine have both been associated with cognitive adverse events and effects on sleep architecture and quality. In contrast, trospium chloride and darifenacin do not appear to be associated with cognitive adverse events and trospium chloride does not negatively affect sleep architecture or quality. Biotransformation by the cytochrome P450 (CYP450) system is an important step in the activation or elimination of a large number of drugs, including oxybutynin, tolterodine, darifenacin and solifenacin, raising the possibility of clinically relevant and potentially serious drug interactions. In elderly patients, such interactions are of particular relevance given the potential for declining activity of certain members of the CYP450 family combined with decreased hepatic blood flow, which can reduce first-pass metabolism and thus the bioavailability of drugs metabolised via this route. Of the antimuscarinic agents used to treat OAB, only trospium chloride is not extensively metabolised in the liver by the CYP450 system and is excreted largely as the active parent compound in the urine. This paper provides an overview of the pathophysiology of OAB and reviews current approaches to achieving a differential diagnosis and selecting appropriate treatment for the older patient. The pharmacology and clinical effects of current medication for the treatment of OAB symptoms in patients defined by the OAB pharmacology literature as 'elderly' are also reviewed. PMID- 16363887 TI - Solifenacin in overactive bladder syndrome. AB - Solifenacin is a bladder-selective, muscarinic (M(1) and M(3)) receptor antagonist. In animal studies, the selectivity of solifenacin for the bladder over the salivary glands was greater than that of tolterodine, oxybutynin, darifenacin or atropine. In large, 12-week, randomised, double-blind, multicentre clinical trials, solifenacin 5 and 10mg once daily improved symptoms of overactive bladder syndrome (OAB) [urinary urgency, frequency, incontinence and nocturia] and increased functional bladder capacity to a significantly greater extent than placebo. Solifenacin 5 or 10mg once daily was noninferior to tolterodine extended release (ER) 4mg daily for improving urinary frequency and had significantly greater efficacy than tolterodine ER for improving other symptoms of OAB (episodes of urgency, incontinence and urge incontinence) and increasing functional bladder capacity. At least half of all patients receiving solifenacin who were incontinent at baseline were continent by study end in the three comparative studies reporting this parameter. Health-related quality of life was significantly improved with once-daily solifenacin 5 or 10mg versus placebo, as assessed in two 12-week double-blind studies; the improvement was maintained during a 40-week extension study. Solifenacin was generally well tolerated; the most frequently reported adverse events were dry mouth, constipation and blurred vision. PMID- 16363888 TI - Solifenacin in overactive bladder syndrome: a viewpoint by Scott Serels. PMID- 16363889 TI - Solifenacin in overactive bladder: a viewpoint by Hashim Hashim. PMID- 16363890 TI - Endovascular repair of abdominal aortic aneurysms with a unibody stent-graft: 3 year results of the French Powerlink Multicenter Trial. AB - PURPOSE: To report the results of a prospective multicenter study on endovascular repair of abdominal aortic aneurysms (AAA) using the bifurcated Powerlink stent graft. METHODS: Between June 2000 and July 2001, endovascular AAA repair using the bifurcated Powerlink stent-graft was performed at 14 centers on 64 patients (61 men; mean age 70 years, range 56-90) fit for open repair. All procedures were attended by a proctor. Data were collected prospectively, and surveillance computed tomograms were reviewed by an independent center. RESULTS: Technical success was achieved in all cases. There was no postoperative death. During a minimum 3-year follow-up (mean 40.6 months, range 1-51), there were 3 (4.7%) stent-graft migrations associated with type I endoleaks. Two (3.1%) of these patients were converted to open repair, and 1 had a proximal extension implanted. No aneurysm rupture occurred, and no stent-graft degradation was observed. Six (9.4%) patients had secondary endoleak (3 type I and 3 type II). The limb occlusion rate was 3.1%. Six (9.4%) patients had secondary procedures. At 3 years, 58 (91.0%) patients were available for follow-up (4 deaths, the 2 conversions). There was a significant decrease between preoperative and 3-year mean AAA diameter (54.6+/-7 versus 47.6+/-10 mm, respectively; p<0.001). Aneurysm sac shrinkage was observed in 32 (55.2%) patients. Twenty-five (43.1%) patients had a stable aneurysm sac diameter. CONCLUSIONS: Endovascular repair using the bifurcated Powerlink stent-graft is safe and effective. Unibody design seems to confer advantages in terms of durability. These results need to be confirmed by longer follow-up and larger series. PMID- 16363891 TI - A variant deployment technique for the powerlink bifurcated endograft. AB - PURPOSE: To report an alternative technique to the dual-lumen catheter for deployment of the Powerlink stent-graft in patients with angulated sacs and calcified aortic bifurcations. A maneuver is also presented to retrieve the delivery system when it is snagged on the stent. TECHNIQUE: After cutdown of the right common femoral artery (CFA), a 9-F introducer sheath is placed percutaneously into the left CFA. A gooseneck catheter is introduced from the right CFA to capture a 0.035-inch hydrophilic guidewire inserted from the left. A 5-F straight catheter is passed over this guidewire from the left to the right CFA. In angulated aneurysm sacs, a 5-F Hunter catheter is introduced from the right femoral access to support a guidewire through the aneurysm to the suprarenal aorta. Then the guidewire is exchanged with a 0.035-inch Amplatz extra stiff wire, and the Hunter catheter is removed. In other cases, a 0.035-inch Amplatz extra stiff guidewire is placed up to the suprarenal aorta. The endograft delivery system is then deployed in the usual manner. A gooseneck snare is also useful in retrieving the delivery system when it is snagged on the stent at the endograft bifurcation. CONCLUSIONS: This variant technique facilitates the deployment of the Powerlink stent-graft when faced with angulated aneurysms or acute and calcified aortic bifurcations. A gooseneck catheter is helpful in retrieving the delivery system's "olive" after endograft placement. PMID- 16363886 TI - Overcoming gaps in the management of asthma in older patients: new insights. AB - Asthma is under-recognised and undertreated in older populations. This is not surprising, given that one-third of older people experience significant breathlessness. The differential diagnosis commonly includes asthma, chronic obstructive pulmonary disease (COPD), heart failure, malignancy, aspiration and infections. Because symptoms and signs of several cardiorespiratory diseases are nonspecific in older people and diseases commonly co-exist, investigations are important. A simple strategy for the investigation of breathlessness in older people should include a full blood count, chest radiograph, ECG, peak flow diary and/or spirometry with reversibility as a minimum. If there are major abnormalities on the ECG, an echocardiogram should also be performed. Diurnal variability in peak flow readings >or=20% or >or=15% reversibility in forced expiratory volume in 1 second, spontaneously or with treatment, support a diagnosis of asthma. Distinguishing asthma from COPD is important to allow appropriate management of disease based on aetiology, accurate prediction of treatment response, correct prognosis and appropriate management of the chest condition and co-morbidities. The two conditions are usually readily differentiated by clinical features, particularly age at onset, variability of symptoms and nocturnal symptoms in asthma, supported by the results of reversibility testing. Full lung function tests may not necessarily help in differentiating the two entities, although gas transfer factor is characteristically reduced in COPD and usually normal or high in asthma. Methacholine challenge tests previously mainly used in research are now also used widely and safely to confirm asthma in clinical settings. Interest in exhaled nitric oxide as a biomarker of airways inflammation is increasing as a noninvasive tool in the diagnosis and monitoring of asthma. Regular inhaled corticosteroids (ICS) are the mainstay of treatment of asthma. Even in mild disease in older adults, regular preventive treatment should be considered, given the poor perception of bronchoconstriction by older asthmatic patients. If symptoms persist despite ICS, addition of long-acting beta(2)-adrenoceptor agonists (LABA) should be considered. Addition of LABA to ICS improves asthma control and allows reduction in ICS dose. However, older people have been grossly under-represented in trials of LABA, many trials having excluded those >or=65 years of age. On meta-analysis, beta(2)-adrenoceptor agonists (both short acting and long acting) are associated with increased cardiovascular mortality and morbidity in asthma and COPD. While the evidence for excess cardiovascular mortality is stronger for short-acting beta(2)-adrenoceptor agonists, it would be prudent to exercise particular care in using beta(2)-adrenoceptor agonists (long acting and short acting) in those at risk of adverse cardiovascular outcomes, including older people. Regular review of cardiovascular status (and monitoring of serum potassium concentration) in patients taking beta(2)-adrenoceptor agonists is crucial. The response to LABA should be carefully monitored and alternative 'add-on' therapy such as leukotriene receptor antagonists (LRA) should be considered. LRA have fewer adverse effects and in individual cases may be more effective and appropriate than LABA. Long-term trials evaluating beta(2) adrenoceptor agonists and other bronchodilator strategies are needed particularly in the elderly and in patients with cardiovascular co-morbidities. There is no evidence that addition of anticholinergics improves control of asthma further, although the role of long-acting anticholinergics in the prevention of disease progression is currently being researched. Older patients need to be taught good inhaler technique to improve delivery of medications to lungs, minimise adverse effects and reduce the need for oral corticosteroids. Nurse-led education programmes that include a written asthma self-management plan have the potential to improve outcomes. PMID- 16363892 TI - Distal embolus protection with an intra-aortic filter during stent-graft repair of a severely atherosclerotic thoracic aortic aneurysm. AB - PURPOSE: To assess the technical feasibility and efficacy of distal embolus protection with an intra-aortic filter during stent-graft repair in a patient diagnosed with a shaggy aorta. TECHNIQUE: In a 75-year-old man with a severely atherosclerotic thoracic aortic aneurysm, stent-graft repair was combined with transposition of the arch branches to the ascending aorta under thoracotomy. A filtration-type embolus protection device with a nitinol basket and polyester fabric was introduced through a 12-F sheath and opened in the supraceliac aorta during the stent-graft procedure; it was safely pulled back into the sheath after the stent-grafts were implanted. The stent-graft repair was successful, and abundant atheromatous debris was captured in the filter. Neither procedure related embolic events nor neurological deficits were observed. CONCLUSIONS: Use of the intra-aortic filter device to prevent distal embolism during thoracic stent-graft repairs may be feasible and efficacious in severely atherosclerotic patients. PMID- 16363893 TI - Fenestrated and branched endografts: assessment of proximal aortic neck fixation. AB - PURPOSE: To investigate proximal fixation characteristics of different aortic endograft designs: a suprarenally placed fenestrated endograft, a modular branched endograft, an infrarenal endograft with suprarenal bare stent fixation, and the gold standard, a conventional hand-sewn anastomosis. METHODS: Ten human cadaveric aortas were obtained at autopsy and transected 20 mm below the renal arteries to mimic an infrarenal aneurysm neck. In random order, the infrarenal, fenestrated, and branched endografts were deployed into the aorta. Using a hydraulic material testing machine, longitudinal load was applied to the distal end of each endograft until migration occurred, thus defining the displacement force (DF). Subsequently, a hand-sewn infrarenal anastomosis was tested in a similar manner. RESULTS: The median DF was 4.67 N (3.82-6.37) for the infrarenal endograft, 9.17 N (8.03- 10.81) for the fenestrated endograft, and 16.95 N (14.78 19.67) for the branched endograft. The differences in DF between the infrarenal and fenestrated endografts and between the fenestrated and branched designs were statistically significant (both p=0.005). The median force to dislodge the graft from the conventional anastomosis was 89.16 N (71.24-105.23). CONCLUSIONS: Suprarenally placed endografts, especially with additional branch grafts, provide improved proximal fixation compared to an infrarenal endograft with suprarenal bare stent fixation. However, none of the tested endografts approached the optimal, time-proven fixation, the hand-sewn anastomosis. PMID- 16363895 TI - Surgical correction of ascending type a thoracic aortic dissection: simultaneous endoluminal exclusion of the arch and distal aorta. AB - PURPOSE: To describe repair of an ascending type A dissection combining an open ascending tube graft with simultaneous great vessel transposition and antegrade deployment of an endoluminal graft across the arch and into the descending thoracic aorta. CASE REPORT: A 50-year-old man was evaluated at an outside hospital and transferred to our service for treatment of an ascending aortic dissection with associated lower extremity ischemia. Imaging identified an aortic dissection extending from the aortic root to the aortic bifurcation and into the right common iliac artery. A hybrid procedure incorporating both open and endovascular techniques successfully repaired the dissection and aneurysm and restored blood flow to the extremity. CONCLUSIONS: Although less invasive procedures are sometimes appropriate for repair in the descending thoracic aorta, surgical correction of an ascending dissection and endoluminal exclusion of the arch and distal aorta may form the basis of future treatment strategies for complex aortic pathologies, possibly eliminating the need for hypothermic cardiac arrest. PMID- 16363894 TI - Infected endoluminal stent-graft: implications for endotension, late endoleaks, and prophylactic antibiotics. AB - PURPOSE: To retrospectively review a case of stent-graft infection that presented as continued aneurysm expansion after endoluminal repair. CASE REPORT: A 75-year old man with an asymptomatic 6.1-cm abdominal aortic aneurysm (AAA) was treated with a Talent stent-graft, which was complicated by postimplantation syndrome. At 1 year, a secondary intervention was performed for migration with type I endoleak. Despite the absence of a demonstrable endoleak thereafter, the AAA continued to expand until it was 9 cm in diameter and symptomatic. At this stage, 6 months after the secondary procedure, the graft was explanted and an axillobifemoral graft inserted. Propionibacterium acnes was cultured from all specimens of thrombus, aortic wall, and graft. The patient recovered and was asymptomatic until his death from myocardial infarction 6 months after discharge. CONCLUSIONS: Stent-graft infection may be a cause of unexplained endotension. Special culture techniques may be required to identify the infecting organism. Prophylactic antibiotics against skin organisms should be considered for all implantations and arterial diagnostic and therapeutic procedures traversing a stent-graft. PMID- 16363896 TI - Directed migration of smooth muscle cells to engineer plaque-resistant vein grafts. AB - PURPOSE: To test the hypothesis that controlled perivascular release of tissue plasminogen activator (tPA) can generate cleaved extracellular matrix (ECM) chemotactic gradients to guide the migration of vascular smooth muscle cells (SMCs) away from the lumen, thereby limiting neointima formation. METHODS: This hypothesis was tested in rabbit models in which the perivascular surface of vein bypass grafts was treated with microspheres releasing tPA (MS-tPA), microspheres containing no drug (MS-blank), or phosphate buffered saline (PBS). Vein graft segments harvested after 7 days were then evaluated for elastin content, proliferating SMCs, intima-to-media (I/M) ratio, and inflammation; late impact on neointima formation was also examined. RESULTS: The 7-day results demonstrated cleaved elastin gradients and proliferating SMCs that assumed a more peripheral distribution in the MS-tPA group than MS-blank and PBS controls (p<0.05). At 28 days, vein grafts treated with MS-tPA showed a mean I/M ratio (0.35+/-0.04) that was 63.5% lower than PBS controls (0.96+/-0.07, p<0.005) and 43.5% lower than MS blank specimens (0.62+/-0.08, p<0.05). CONCLUSIONS: Perivascular release of tPA modifies ECM gradients, directionally guides SMC migration away from the lumen, and limits neointima formation. PMID- 16363897 TI - Long-segment SFA stenting--the dark sides: in-stent restenosis, clinical deterioration, and stent fractures. AB - PURPOSE: To determine and compare the rates of in-stent restenosis, late clinical deterioration, and stent fractures in nitinol stents versus Wallstents implanted for suboptimal angioplasty in the superficial femoral artery (SFA). METHODS: Interrogation of an angioplasty database identified 286 consecutive patients (178 men; mean age 67+/-10 years, range 44-87) with severe claudication (n=254) or critical limb ischemia (n=32) who had stents implanted after suboptimal angioplasty over a 5-year period. Wallstents with a mean stented lesion length of 107+/-71 mm were implanted in 116 patients, while nitinol stents were used in 170 patients: 45 SMART stents (mean stented lesion length 139+/-88 mm) and 125 Dynalink/Absolute stents (mean stented lesion length 125+/-84 mm). Patients were followed for in-stent restenosis (>50%) by duplex ultrasound, clinical deterioration by at least 1 Fontaine stage compared to baseline, and stent fractures by biplanar radiography. RESULTS: In-stent restenosis rates at 1, 2, and 3 years were 46%, 66%, and 72% for Wallstents compared to 20%, 36%, and 53% for nitinol stents (p<0.001), respectively, without significant difference between the 2 nitinol stent groups (p=0.59). Clinical deterioration at 1, 2, and 3 years was found in 10%, 15%, and 18% with Wallstents versus 4%, 5%, and 5% with nitinol stents (p=0.014), respectively, without difference between the 2 nitinol stent groups (p=0.47). Fracture rates were 19% for Wallstents after a mean 43+/ 24 months, 28% for SMART stents after mean 32+/-16 months, and 2% for Dynalink/Absolute stents after a mean 15+/-9 months. CONCLUSIONS: Intermediate term in-stent restenosis remains a major problem even with current nitinol stent technology; however, clinical deterioration seems no matter of serious concern with SMART and Dynalink/Absolute stents. Stent fractures may be lower with Dynalink/Absolute stents, but randomized head-to-head comparisons are needed to validate these data. PMID- 16363898 TI - Sirolimus-eluting versus bare stents for bailout after suboptimal infrapopliteal angioplasty for critical limb ischemia: 6-month angiographic results from a nonrandomized prospective single-center study. AB - PURPOSE: To report the 6-month angiographic results from a prospective single center study investigating the efficacy and outcome of sirolimus-eluting stents used for bailout after infrapopliteal revascularization of patients with critical limb ischemia (CLI). METHODS: Twenty-nine patients (21 men; mean age 68.7 years) underwent infrapopliteal revascularization with bare metal stents (group B) implanted for bailout in 65 lesions (38 stenoses and 27 occlusions) in 40 infrapopliteal arteries. Another 29 patients (21 men; mean age 68.8 years) underwent infrapopliteal bailout stenting with sirolimus-eluting stents (group S) in 66 lesions (46 stenoses and 20 occlusions) in 41 vessels. Preliminary 6-month angiographic and clinical results were analyzed. RESULTS: Hyperlipidemia and symptomatic cardiac and carotid diseases were more pronounced in group S (p<0.05). Technical success was 96.6% (28/29 limbs) in group B versus 100.0% in group S (p=0.16). Six-month primary patency was 68.1% in group B versus 92.0% in group S (p<0.002). Binary in-stent and in-segment restenosis rates were 55.3% and 66.0%, respectively, in patients with bare stents versus 4.0% and 32.0%, respectively, in patients treated with the sirolimus-eluting stents (both p<0.001). The target lesion re-intervention rate at 6 months was 17.0% in group B versus 4.0% in group S (p=0.02). Limb salvage was 100% in both groups. Six-month mortality and minor amputation rates were 6.9% and 17.2%, respectively, in group B versus 10.3% and 3.4%, respectively, in group S (p=0.32 and p=0.04, respectively). CONCLUSIONS: Sirolimus-eluting stents seem to restrict neointimal hyperplasia in the infrapopliteal vascular bed. PMID- 16363899 TI - Does stent overlap influence the patency rate of aortoiliac kissing stents? AB - PURPOSE: To determine if the position of kissing stents in the distal aorta has any influence on the patency rate. METHODS: A retrospective review was conducted of 41 patients (22 men; median age 60.8 years, range 44-86) electively treated for atherosclerotic aortoiliac occlusive disease with angioplasty and kissing stents between January 1997 and January 2005. Two patient groups were defined by reviewing postinterventional anteroposterior radiograms: (1) patients in whom the proximal end of the kissing stents overlapped more than half of their angiographic width within the aorta ("crossing" group) and (2) patients in whom the proximal ends of the stents overlapped half of their width or less ("non crossing" group). RESULTS: At 2 years, the primary and assisted primary patency rates by life-table analysis were 60.8% and 69.4%, respectively, for the 35 patients included in the life-table analysis. There was no significant difference between the 16-patient "crossing" group and the 19-patient "non-crossing" group in terms of the baseline demographic, morphological, and procedural variables. The primary and assisted primary patency rates at 2 years for the "non-crossing" group were significantly higher (94.1% and 100%, respectively) compared to 33.2% and 45.3%, respectively, for the "crossing" group (p=0.01). CONCLUSIONS: Failure of kissing stents in the aortic bifurcation may be significantly increased by the overlap of the free proximal stent ends in the distal aorta. PMID- 16363900 TI - Below-the-knee angioplasty in patients with end-stage renal disease. AB - PURPOSE: To determine clinical efficacy of below-the-knee (BTK) angioplasty in patients with end-stage renal disease (ESRD). METHODS: Interrogation of a prospectively maintained database containing 2,659 patients treated at a tertiary referral hospital between February 1995 and June 2004 identified 29 ESRD patients (21 men; median age 69 years, IQR 10.12) who had 73 infrapopliteal atherosclerotic lesions treated in 38 ischemic limbs. The indication for treatment was intermittent claudication in 13 (34%) and critical limb ischemia in 25 (66%) limbs. BTK angioplasty was attempted either alone (n=18) or combined with an endovascular inflow procedure (n=20). Primary clinical success was defined as hemodynamic improvement (ABI increase >or=0.1) and/or symptomatic improvement (at least one clinical category). Cumulative rates were calculated according to the Kaplan-Meier estimate. RESULTS: Primary technical success reached 97%, whereas hemodynamic improvement was obtained in only 50% (19/38) of the limbs treated. The pedal arteries were severely diseased in all, and complete occlusion of the pedal arch was found in 58% (18/31) of limbs on completion angiography. Median follow-up was 5.9 months (IQR 11.5). Primary clinical success was 17%, 11%, 11%, and 11% in patients with BTK angioplasty alone and 53%, 45%, 45%, and 45% in patients with inflow procedures after 3, 6, 9, and 12 months, respectively (p=0.017). Limb salvage was 73% at 12 months. Subgroup analyses showed significantly better clinical results in men (p=0.003) and in patients on hemodialysis compared to peritoneal dialysis (p=0.037). CONCLUSIONS: Clinical efficacy of BTK angioplasty is limited in patients with ESRD because of the severely diseased pedal arteries. Further studies are warranted to define subgroups of patients likely to experience a more favorable outcome. PMID- 16363901 TI - Ultrasound-guided retrograde popliteal artery catheterization: experience in 174 consecutive patients. AB - PURPOSE: To assess the safety and efficacy of ultrasound (US) guidance during retrograde popliteal artery catheterization (RPAC) and identify the risk factors for late hemostasis and complications. METHODS: A retrospective review was undertaken of consecutive patients who underwent grayscale US-guided RPAC during the last 8 years. Chart review identified 174 patients (150 men; mean age 61+/-10 years, range 37-84) with 247 iliofemoral lesions, which were treated via US guided RPAC. Complications were classified as puncture-related versus angioplasty related and major versus minor. Risk factors for complications and late hemostasis were evaluated with logistic and linear regression analyses, respectively. RESULTS: All 234 US-guided RPACs were technically successful. No arteriovenous fistula (AVF) or dissection/thrombosis of the popliteal artery was observed. The mean time to hemostasis was 6.9+/-2.3 minutes. The presence of femoral stenosis (versus occlusion) and use of large (6 to 7-F) sheaths were significant risk factors for late hemostasis. In 234 procedures, 15 (6.4%) complications developed; 10 (4.3%) were puncture-related (3 major, 7 minor) and 5 (2.1%) were angioplasty-related (3 major, 2 minor). Intra-arterial fibrinolysis was found to be the only significant risk factor for overall and puncture-related complications. CONCLUSIONS: Our results suggest that US-guided RPAC is at least as safe as other RPAC methods described in the literature. In contrast to generally held concerns, hemostasis is easy to obtain, and multiple punctures and the use of large sheaths appear safe. These results should be taken into consideration during the selection of an access site for endovascular treatment of superficial femoral artery and tandem iliofemoral lesions. PMID- 16363902 TI - Endovascular brachytherapy after femoropopliteal balloon angioplasty fails to show robust clinical benefit over time. AB - PURPOSE: To determine if the short-term efficacy of adjunctive endovascular brachytherapy (EVBT) is maintained over time in patients undergoing balloon angioplasty (BA) of femoropopliteal atherosclerotic lesions. METHODS: To evaluate the long-term clinical and angiographic outcome of EVBT, 147 consecutive patients (82 men; mean age 70.8+/-8.5 years) with 147 treated limbs were randomized to BA with (n=72, 49%) or without (n=75, 51%) adjunctive EVBT (12 or 14-Gy from an (192)Ir source, no centering, a 5-mm reference depth). Sixty-eight (46%) limbs were treated for de novo and 79 (54%) for recurrent femoropopliteal lesions. Clinical follow-up at 1, 3, 6, and 12 months and annually thereafter included evaluation of symptoms, ankle-brachial index (ABI), and intra-arterial angiography for new/worsening symptoms or at follow-up between 2 and 5 years. Sustained clinical success was defined as improvement in ABI >or=0.1 and/or of symptoms without repeated target lesion revascularization. Angiographic restenosis was defined as >or=50% diameter reduction. Subgroup analysis was performed for de novo versus recurrent lesions. RESULTS: Mean clinical follow-up was 32.3+/-21.5 months. Angiographic follow-up was available in 83 (56%) patients (41 BA and 42 BA+EVBT) at a mean 31.8+/-20.7 months. Cumulative sustained clinical success rates at 1, 2, and 3 years, respectively, were 84.3%, 82.1%, and 76.4% after BA versus 82.4%, 69.8%, and 67.5% after BA+EVBT (p=0.26 by log-rank). Although the proportion of patients undergoing follow-up angiography was moderate, the freedom from angiographic restenosis at 1, 2, and 3 years was 70.7%, 63.1%, and 47.1% after BA versus 82.7%, 64.3%, and 64.3% after BA+EVBT (p=0.16 by log-rank). No differences were found between BA and BA+EVBT outcomes in patients with de novo versus recurrent femoropopliteal lesions. CONCLUSION: The seemingly beneficial short-term effects of BA+EVBT are not sustained in the longer term, with no robust clinical improvement after angioplasty of atherosclerotic de novo or recurrent femoropopliteal lesions at up to 5 years. PMID- 16363903 TI - Is there recanalization of the great saphenous vein 2 years after endovenous laser treatment? AB - PURPOSE: To report the 2-year single-center results of endovenous laser treatment (EVLT) for reflux in the great saphenous vein (GSV). METHODS: From January 2002 to January 2003, 85 symptomatic patients (56 women; mean age 49 years, range 27 80) underwent EVLT in 100 limbs. All patients were symptomatic, and the majority (67, 79%) had CEAP clinical class C2 venous disease. After treatment, they were monitored by clinical evaluation and duplex imaging. RESULTS: The initial treatment was completed in 93 limbs. Complications consisted of bruising (31%), tightness (17%), pain (14%), induration (2%), and superficial thrombophlebitis (2%). No severe complications were observed. Over a mean follow-up of 29 months (range 24-37), 3 patients died and 14 were lost to follow-up, leaving 88 (95%) and 76 (82%) limbs available for imaging surveillance at 1 and 2 years, respectively. At 3 months, treatment was anatomically successful in 84% of cases (78 complete occlusion, 7 partial occlusion, and 8 nonocclusion) and functionally successful in 89% (83 no reflux, 10 reflux). All technical failures and 73% (n=11) of the treatment failures occurred in the first half of the studied population, indicating a learning curve effect (p=0.015). Mean energy delivered per unit length was 39+/-8 J/cm (range 25-65) for successful treatment (n=78) and 30+/-10 J/cm (range 21-50) for failed treatment (n=15). No recanalization or recurrent GSV reflux after anatomically and functionally successful treatment was observed in 73 and 61 limbs at 1 and 2-year follow-up, respectively. CONCLUSIONS: EVLT is a feasible, safe, and fast procedure for eliminating GSV reflux and has excellent cosmetic results. Despite the learning curve, we believe that the treatment results are promising. When successful treatment is achieved by EVLT, a prospective follow-up of 2 years demonstrates durable results. PMID- 16363904 TI - Simultaneous superficial venous surgery and on-table subintimal angioplasty in the treatment of venous leg ulcers with arterial compromise. AB - PURPOSE: To determine the feasibility of synchronous superficial venous surgery and on-table subintimal angioplasty in the treatment of venous ulcers with arterial compromise. METHODS: Between January 1992 and December 2004, 9 patients (8 women; median age 83 years, range 72-92) underwent the synchronous procedure. No patient had deep venous reflux; 4 patients presented with rest pain and 2 were diabetic. Their medical records were retrospectively reviewed to gather data on clinical assessment, ankle-brachial index (ABI), lower limb arterial and venous duplex scans, and treatment variables. RESULTS: Angioplasty was technically successful (median ABI improvement 0.31, range 0.23-0.34) in all procedures, which lasted a median 85 minutes (range 60-160). The procedure was performed under general anesthesia in 7 patients, local in 1, and spinal in 1. Superficial venous surgery was performed first in 8 patients: 6 had great saphenous vein (GSV) ligation without stripping, followed by subintimal angioplasty, and 1 also had small saphenous vein (SSV) ligation. One patient underwent SSV ligation first, followed by angioplasty; another had GSV disconnection and stripping, followed by angioplasty. In 1 patient, angioplasty preceded GSV surgery. One patient required repeat angioplasty 1 week later for re-occlusion. All ulcers healed (median 4 months, range 1-36). Five patients died during a median 32-month follow-up (range 4-82) from unrelated causes; there was 1 ulcer recurrence at 3 years in a patient who declined further investigation and treatment. CONCLUSIONS: Simultaneous superficial venous surgery and on-table subintimal angioplasty is a safe and novel strategy in the management of a select subgroup of patients with venous ulcers and coexistent arterial compromise. PMID- 16363905 TI - False aneurysm formation following cutting balloon angioplasty in the renal artery of a child. AB - PURPOSE: To report the fortuitous discovery of a false aneurysm 7 years after cutting balloon angioplasty for severe renal artery fibromuscular dysplasia in a child. CASE REPORT: A 3-year-old girl with neurofibromatosis was referred to our institution because of high blood pressure (220/160 mmHg). Computed tomography identified coarctation of the aorta and severe bilateral renal artery stenoses. The coarctation was successfully resected. One month later, bilateral renal artery angioplasty with a 3-mm balloon was unsatisfactory, so a second angioplasty one month later was done with a 3.25-mm cutting balloon. This procedure was complicated by a minimal arterial rupture, which resolved spontaneously after inflation of a regular balloon. Normal blood pressure was restored. The child was lost to follow-up until 7 years later, when recurrent hypertension (200 mmHg systolic) prompted referral again. Arteriography showed a very severe stenosis on the right side and a 30-mm false aneurysm of the left renal artery at the rupture site. Due to her age, the patient underwent surgery, which brought the blood pressure under control. CONCLUSIONS: False aneurysm of the renal arteries is a rare complication of percutaneous angioplasty. In a child, the cutting balloon would appear to be contraindicated for concentric dysplastic stenoses that are resistant to regular balloon angioplasty. PMID- 16363906 TI - "Kissing aneurysms" of the anterior communicating artery treated with coil embolization. AB - PURPOSE: To present a case of kissing aneurysms of the anterior communicating artery treated with endovascular coil embolization and discuss the advantages and disadvantages of this technique compared with neck clipping. CASE REPORT: A 48 year-old man became drowsy and was admitted to the hospital; he had right hemiparesis and aphasia. Computed tomography revealed diffuse subarachnoid hemorrhage; diagnostic angiography identified an aneurysm at the left A1-A2 junction of the anterior communicating artery and another in the distal anterior cerebral artery (ACA). Endovascular coil embolization was performed on the same day. During the procedure, the 3-mm-diameter junctional aneurysm was successfully packed with coils, but an additional aneurysm was suspected; right carotid angiography following embolization of the left aneurysm clearly showed a mirror image aneurysm of the right A1-A2 junction. The right aneurysm was treated using the same technique. The broad-necked ACA aneurysm was unsuitable for embolization, so neck clipping was performed 5 weeks later. The patient was discharged to his home following complete recovery 7 weeks after the coil embolization. CONCLUSIONS: Kissing aneurysms are a rare and specific type of multiple aneurysms that require caution in diagnosis and surgical management. Endovascular treatment may be suitable because it does not involve dissection around the aneurysms. PMID- 16363907 TI - Regarding "Endovascular AAA repair with the aortomonoiliac endofit stent-graft: two years' experience". PMID- 16363908 TI - The global campaign to eliminate leprosy. PMID- 16363909 TI - Thyrotoxicosis and pregnancy. PMID- 16363910 TI - Moral imagination: the missing component in global health. PMID- 16363911 TI - Selection bias in meta-analyses of gene-disease associations. PMID- 16363912 TI - Tweaking microtubules to treat scleroderma. PMID- 16363913 TI - Breaking up (amyloid) is hard to do. PMID- 16363914 TI - Controlling the Spread of HIV/AIDS in the Indian Subcontinent. PMID- 16363915 TI - The statistical significance of suffering. PMID- 16363916 TI - Correction/clarification about FDA review documents. PMID- 16363918 TI - Improving health by investing in medical education. PMID- 16363920 TI - Influences on the feeding behavior of three mammals in the Maruyama Zoo: bears, elephants, and chimpanzees. AB - Modifications of feeding conditions are essential to the establishment of environmental enrichment in zoos. In this study, we attempted to increase the duration of feeding by varying feeding conditions, such as the spatial or temporal distribution of foods, while keeping the sorts and amounts of food the same. Subjects included nonhuman animals reared at the Maruyama Zoo in Sapporo, Japan: 3 bears (Ursus arctos), 2 elephants (Elephas maximus), and 5 chimpanzees (Pan troglodytes). Considering that the feeding ecologies of these animals differed a great deal from those in the wild, we used the focal animal sampling method. Consisting of the spatial dispersive and massed feeding conditions, the experiment with bears found feeding time increased more in the dispersive condition. In addition, the behavioral rhythm of alternate feeding and sleeping appeared. The experiment with elephants, consisting of the temporal dispersive and massed condition, increased feeding time under the massed condition. The experiment using chimpanzees reversed conditions used in the elephant study and increased feeding time under the dispersive condition. Results indicated that these simple modifications both influenced an increasing duration of feeding and affected behavioral patterns in a day. PMID- 16363919 TI - The evolution of senescence and post-reproductive lifespan in guppies (Poecilia reticulata). AB - The study of post-reproductive lifespan has been of interest primarily with regard to the extended post-menopausal lifespan seen in humans. This unusual feature of human demography has been hypothesized to have evolved because of the "grandmother" effect, or the contributions that post-reproductive females make to the fitness of their children and grandchildren. While some correlative analyses of human populations support this hypothesis, few formal, experimental studies have addressed the evolution of post-reproductive lifespan. As part of an ongoing study of life history evolution in guppies, we compared lifespans of individual guppies derived from populations that differ in their extrinsic mortality rates. Some of these populations co-occur with predators that increase mortality rate, whereas other nearby populations above barrier waterfalls are relatively free from predation. Theory predicts that such differences in extrinsic mortality will select for differences in the age at maturity, allocation of resources to reproduction, and patterns of senescence, including reproductive declines. As part of our evaluation of these predictions, we quantified differences among populations in post-reproductive lifespan. We present here the first formal, comparative study of the evolution of post-reproductive lifespan as a component of the evolution of the entire life history. Guppies that evolved with predators and that experienced high extrinsic mortality mature at an earlier age but also have longer lifespans. We divided the lifespan into three non-overlapping components: birth to age at first reproduction, age at first reproduction to age at last reproduction (reproductive lifespan), and age at last reproduction to age at death (post-reproductive lifespan). Guppies from high-predation environments live longer because they have a longer reproductive lifespan, which is the component of the life history that can make a direct contribution to individual fitness. We found no differences among populations in post-reproductive lifespan, which is as predicted since there can be no contribution of this segment of the life history to an individual's fitness. Prior work on the evolution of post reproductive lifespan has been dominated by speculation and correlative analyses. We show here that this component of the life history is accessible to formal study as part of experiments that quantify the different segments of an individual's life history. Populations of guppies subject to different mortality pressures from predation evolved differences in total lifespan, but not in post reproductive lifespan. Rather than showing the direct effects of selection characterizing other life-history traits, post-reproductive lifespan in these fish appears to be a random add-on at the end of the life history. These findings support the hypothesis that differences in lifespan evolving in response to selection are confined to the reproductive lifespan, or those segments of the life history that make a direct contribution to fitness. We also show, for the first time, that fish can have reproductive senescence and extended post reproductive lifespans despite the general observation that they are capable of producing new primary oocytes throughout their lives. PMID- 16363921 TI - Elephants in Thailand: determinants of health and welfare in working populations. AB - The Asian elephant (Elephas maximus) has played a prominent role in Thai history and society. However, in the face of modernization, elephant handlers have been struggling to justify their continued ownership. As a result, working elephants may still encounter situations in which their health and welfare are jeopardized. This study developed both a survey instrument and a visual assessment to describe and evaluate the health and living conditions of elephants encountered in a variety of work and living situations. These situations were found to be significantly associated with whether or not an elephant received proper husbandry or was in good body condition. These results may prove valuable in predicting the welfare of elephants according to work and living situations. PMID- 16363922 TI - The nuts and bolts of captive chimpanzee diets and food as enrichment: a survey. AB - Beyond planning for good nutrition, one of the Cornerstones of a successful captive-chimpanzee wellness program is the opportunity for the nonhuman primates to express normal feeding and foraging behaviors. This study presents the results of a survey of 11 institutions-8 zoological and 3 research facilities-given to ascertain how daily diets and feeding enrichments were accomplished. Information represents 806 captive chimpanzees, and entails a composite of captive chimpanzee daily diets, forage and browse materials, periodic treats, medicating and supplemental foods, alternative feeding techniques, and food devices. Results suggest facilities are moving away from timed feeding schedules, which tend to increase prefeeding levels of agonism, to feeding a wide variety of foods throughout the day and using a wide variety of feeding techniques. PMID- 16363923 TI - Beyond overpopulation: a comment on Zawistowski et al. and Salman et al. AB - The intentional production and destruction each year of millions of companion animals is a sobering fact. The need for meaningful statistical data on this phenomenon is urgent. No less pressing, however, is the need for a conceptual framework to make sense of the empirical findings. The conventional focus on overpopulation is inadequate and places unnecessary limits on the way in which the problem is approached. I propose in its place a market-based model that takes seriously the role of consumer demand and considers the flaws and inefficiencies distorting the market for companion animals. Shifting attention to the choices consumers make in acquiring and discarding animal companions suggests new possibilities for change and provides a useful focal point for further empirical work. PMID- 16363924 TI - A method to determine when active translocation of nonhuman primates is justified. AB - This article addresses translocation, or artificial dispersal, the movement of one or more organisms from one location to another, and focuses on the decisions to be made before translocation begins. Scientific, economic, or pragmatic reasons such as pest removal or conservation of biodiversity may account for undertaking translocation. When is translocation ethical, and how can that decision be determined? This article provides one quantitative and utilitarian method for evaluating these questions. Although this analysis may apply to any nonhuman species for which costs and benefits can be assessed, the examples in this article derive from the nonhuman primate literature. PMID- 16363925 TI - Jinxed lynx? PMID- 16363928 TI - Is there a future for zoos? PMID- 16363929 TI - Marine mammal welfare: an exploration of the world of marine mammals. PMID- 16363930 TI - Pain management and humane endpoints. PMID- 16363931 TI - Deprivation and enrichment in laboratory animal environments. PMID- 16363932 TI - Environmental enrichment for laboratory rodents: animal welfare and the methods of science. AB - Because of the difficulty of' establishing objective measures of laboratory rodents' psychological well-being, developing environmental enrichment programs that are actually beneficial to rodents destined to participate in laboratory research is particularly challenging. Many studies of effects of environmental complexity, social housing, and increases in cage size suggest that professional judgments as to the impact of diverse types of environmental enrichment on rodent welfare are not a reliable basis for evaluating the outcomes of enrichment programs for laboratory rodents. Successful enrichment programs will vary from one rodent species to another, between sexes, as well as between age classes. There is a need for objective, measurable goals for proposed environmental enrichment programs for rodents, as well a s for empirical investigations of the beneficial and detrimental consequences of proposed environmental manipulations. PMID- 16363933 TI - Kennel enrichment: exercise and socialization of dogs. AB - In the last 50 years, there has been a growing need for storage and management systems for the production and maintenance of large numbers of dogs. Unwanted dogs and strays, detained in kennels, stay for various lengths of time. Large kennels also produce dogs for sale as companion animals, for the service dog industry (police and guide dogs), for biomedical research, and for use by dog food companies. Across the United States, literally tens of thousands of dogs are born in kennels and spend their lives in kennels. The laboratory dog, the kennel dog, the service dog, and the companion dog are in an evolutionary transition period, accompanied by concomitant adaptation to stresses signaled by a high frequency of genetic disease and behavioral abnormalities. For kennel enrichment programs, such as socialization and exercise, the modern kenneled dog is a genetically moving target. Specific recommendations apply neither to all breeds nor to the variations within a single breed. PMID- 16363934 TI - Psychological well-being of nonhuman primates: a brief history. PMID- 16363935 TI - Pet overpopulation: data and measurement issues in shelters. AB - Data collection and analysis within animal shelters are critical to developing effective programs that reduce the number of dogs and cats euthanized each year. However, current data collection efforts are insufficient to identify the magnitude, dynamics, or root causes of euthanasia in animal shelters across the United States. The purpose of this study was to examine potential solutions to the underlying root causes of pet overpopulation, with 2 elements. The first, more explicit goal was to establish a baseline of shelter data, policies, and viewpoints through a detailed survey of 186 shelters, 12 site visits, and numerous interviews. The findings suggest large variation in local issues faced by shelters, as well as a nearly universal focus on sterilization as a solution. The greater objective, however, was to use this information as an impetus to improve the process by which shelters amalgamate information and effectively use it to target the most pressing needs within their communities. We believe the essential step is to provide shelters with an analytical tool that would yield informational benefits exceeding the cost of data collection. Such an improvement would have a positive spillover effect on researchers, donors, and others attempting to collect standardized, geographically scalable data. This article presents an overview of the survey findings, as well as a prototype of a tool to help improve data amalgamation and analysis efforts within shelters. PMID- 16363936 TI - Preliminary evaluation of porcine zona pellucida (PZP) immunocontraception for behavioral effects in feral horses (Equus caballus). AB - Successful management of captive populations of wild animals requires effective control of reproduction. Contraception is one tool for controlling reproduction of animals in zoos; however, the options available to the animal manager are limited. Contraceptives vary in efficacy, reversibility, and side effects, and thus may not be suitable for widespread use. One consideration when selecting a contraceptive is its potential for side effects on behavior, especially given the fact that reproduction plays such a prominent role in the biology of any species. To date, there have been few evaluations of contraceptives for behavioral effects, and those that have been conducted have focused on hormone-based contraceptives. This study sought to evaluate a novel method of population control, immunocontraception, for behavioral effects in a population of feral horses. Porcine zona pellucida (PZP) immunocontraception prevents fertilization of ova and does not alter normal hormone secretion patterns. It therefore should leave the animal behaviorally intact in terms of reproductive behavior. The study examined the behavior of 43 sexually mature mares on Assateague Island during the 1997 breeding season and, with help from Earthwatch volunteers, collected observations over a 3-month period. The study found no significant differences between treated and untreated mares in general activity budget, aggression given or received, and spatial relationships relative to the stallion. These preliminary findings indicate that PZP contraception seems to have no acute behavioral effects on the behavior of individuals. The study findings also suggest that PZP could be a desirable and effective management tool for captive species in which social behavior plays an integral role in group dynamics. Analyses of group level effects and population level effects are continuing. PMID- 16363937 TI - Responsible animal-based research: three flags to consider. AB - Flagging experiments is one way to reduce the amount of nonhuman animal-based research. Experiments flagged for the following 3 characteristics are highly questionable: few or no citations received in the years following publication, thus having little or no influence on subsequent research; large numbers of animals used; and invasive procedures, such as injections or anesthesia and surgery, used on animals who were then allowed to recover enough to be tested in some way. Selecting and analyzing a sample of articles describing such experiments (such as those by authors at Canadian universities) determines which journals, author-affiliated universities, and acknowledged funding sources have the most flags. One could also assess the work of individual scientists. Using recommendations supplied, efforts to reduce the suffering of animals and the waste of tax dollars can then focus especially on these entities. PMID- 16363938 TI - Animals in elementary school education in California. AB - The purpose of this project was to provide preliminary, descriptive data concerning the uses of nonhuman animals in northern California elementary school classrooms. The project comprised a mail survey that was sent to all elementary school teachers in Stanislaus County, California, followed by a workshop with survey respondents. Information gathered included that about animals and the use of animals, the learning objectives for the animal-related instructional activities, and the kinds of resources that would help teachers to improve or expand their existing animal-related activities or to create new ones. Most of the teachers were keeping either live animals or a variety of nonliving specimens in the classroom. They reported that animals and information about animals are popular and effective foci of instruction throughout the primary grades. The majority of respondents stated that live animals, in addition to providing formal lessons in science and cross-curricular instruction, are important for teaching children humane values. They also reported the many drawbacks of maintaining live classroom animals, such as the responsibility and cost of ongoing care, potential health and safety issues for the children, and concerns about the welfare of the animals. Because animals used in precollege education are not subject to legal regulations in the United States, some degree of oversight by the school or school district may help to avert potential problems. PMID- 16363939 TI - The International Conference on the use of humane endpoints in animal experiments for biomedical research. PMID- 16363940 TI - Companion dog temperament traits. AB - Owners rated their companion dogs (Canis faamiliarus) on the frequency of occurrence of 127 descriptions of behavior. We conducted an exploratory principal components analysis of these behaviors for 2,018 participants that produced 22 interpretable factors (or traits). Four related to aggression indicate that aggression is not a unitary concept. Three factors relate to play and 3 describe vocal behavior. Regression analyses examined the relation of sex and obedience training (OT) to factor scores. Some unexpected relations between OT and factor scores were identified. Frequency distributions for factor scores were positively skewed for undesirable traits and negatively skewed for more desirable traits. Some common beliefs about companion dog behavior problems, such as the relation between playing tug-of-war games and aggression, are challenged. The identified factors provide some insight into the structure of temperament in companion dogs and represent traits that can be measured for individual dogs, although further refinement is needed. PMID- 16363941 TI - Effects of selected behavioral enrichment devices on behavior of Western Lowland Gorillas (Gorilla gorilla gorilla). AB - Environmental complexity plays an integral role in the activity and psychological well-being of primates. The experiment described in this article evaluates the effects of nonintrusive, inexpensive, and easily managed behavioral enrichment devices on the behavior of a group of captive Western lowland gorillas. Devices used included cardboard boxes containing food items, paper bags containing food items, burlap rags, and willow and maple browse. The enrichment devices increased foraging, social play, and solitary play behaviors. Sedentary behaviors decreased. Rags, bags, browse, and boxes did not statistically decrease the incidence of regurgitation/reingestation or coprophagy. Depending on the type of enrichment item used, the effects on agonism and manipulation of enrichment item were variable. To make informed management decisions about the psychological well being of captive animals, it is important to objectively quantify and examine the influences on their behavior. PMID- 16363942 TI - Animal care courses: helping fulfill the mandate of animal care committees in Canada. AB - Animal Care Committees (ACCs) at Canadian universities and research centers operate under the aegis of the Canadian Council on Animal Care (CCAC) and its guidelines for the humane care and treatment of animals in teaching, research, and testing. All Canadian universities have at least one active committee. The committees are expected to assume an educative role beyond the provision of information concerning housing, maintenance, and appropriate conditions for the treatment of animals in research. This includes critical examination of the serious ethical issues involved in animal research within the context of the principles and practices endorsed by the CCAC. One-day animal care courses provided by ACCs at three Canadian universities are described. Comparisons are made between the content and structure of curricula and the ways these relate to the teaching and research mandate in each institution, focusing particularly on the teaching of ethics in each course. The implications for heightening awareness of ethical issues in animal research and improving the effectiveness of these courses are discussed. PMID- 16363943 TI - Bovine spongiform encephalopathy statement of possible relation with new variant Creutzfeldt-Jakob disease: effects on the welfare of United kingdom cattle. AB - Although measures to control bovine spongiform encephalopathy (BSE) had been in force in the United Kingdom for many years and had resulted in a marked decline in clinical cases, the announcement by the Secretary of State for Health on March 20, 1996, that a new variant form of Creutzfeldt-Jakob Disease may be linked with exposure to BSE, resulted in the introduction of several new control measures. These measures included a scheme banning human consumption of meat from cattle who were more than 30 months old (the so-called "over 30-month scheme;" OTMS), a subsidy for slaughter of calves, and additional inspections of abattoirs. The altered slaughter procedures and lack of rendering facilities meant an initial backlog of OTMS animals having to remain on farms. This placed pressures on accommodation and feed stocks, the latter being in short supply because of a poor grass growth the previous summer. Initially, the long delay before the removal of casualty animals from the farm resulted in increased summer mastitis problems for nonlactating cows. The export of calves to the European Union for veal production was banned, thereby allowing the cessation of a previously legal trade that was considered distasteful by many members of the general public. Financial concerns disturbed and continue to disturb affected farmers. PMID- 16363944 TI - Commentary on de Cock Buning: a United kingdom perspective. AB - de Cock Buning (1998) highlighted the existence of alternative and more favorable options available to xenotransplantation. Clearly, there is a need to emphasize a review of existing organ procurement programs worldwide. A tree interest in the welfare of animals encourages increased liaison between transplant communities throughout the world to discuss the experiences of various procurement programs. PMID- 16363945 TI - Commentary on de Cock Buning: animal welfare problems in xenotransplantation. PMID- 16363947 TI - EuroNICHE 10th Anniversary Conference. PMID- 16363948 TI - Moving: characteristics of dogs and cats and those relinquishing them to 12 U.S. animal shelters. AB - The National Council on Pet Population Study and Policy Regional Shelter Survey identified moving as the most often cited of 71 reasons for relinquishing dogs and the 3rd most common reason for relinquishing cats. Most relinquished companion animals were less than 3 years old and female. Dogs were most often intact, whereas cats were most often altered. Friends had given the majority of these companion animals to the relinquishers, who had obtained them at no cost. Most had lived with the relinquishers for less than 2 years. The majority of relinquishers were White and female, and had at least a high school education. Based on the U.S. population age distribution, young adults seem to be using shelters at a significantly higher rate than would be expected. These age groups are also more mobile, according to U.S. Census data. Therefore, educational efforts that target young, potentially mobile adults could decrease the number of animals relinquished. PMID- 16363949 TI - Behavioral indexes of poor welfare in laboratory rats. AB - A study compared the behavior of rats kept under standard conditions with the behavior of those from enriched, semi-enriched, and single-housed conditions to determine whether standard conditions have detrimental effects on nonhuman animal welfare. The subjects were 35 female Hooded Norway rats. Using an emergence box, open field, and Hebb-Williams maze, the study measured the subjects' behaviors. Housing conditions significantly affected the behavior of the rats as measured by all 3 procedures. The results of the rats from standard conditions were intermediate between those of the single-housed and the more enriched groups. Thus, enriching the standard environment would ameliorate behavioral deficits in rats kept under standard conditions. PMID- 16363950 TI - Effect of castration on rabbits housed in littermate pairs. AB - Conventional solitary caging for research rabbits precludes many postural and social behaviors. Several studies that have investigated enrichment strategies for rabbit does have found intact males usually too aggressive for paired or group housing. This study investigated the possibilities that male siblings will remain compatible if never separated from birth and that castration will improve compatibility among young sibling male rabbits. As a side benefit of pair housing for all rabbits, the larger cages (modified dog cages) allowed rest and other postures not possible in conventional single cages. In addition, there was no evidence of the stereotypies observed among individually housed rabbits. Castrated, post-weanling, male, Flemish Giant x Giant Chinchilla rabbit littermates demonstrated a significantly higher level of living together compatibly than did the intact controls. All rabbits chose to rest in close proximity or contact, although this preference was stronger in castrated males. During puberty, intact males displayed more serious aggression. We conclude that young adult littermate rabbits can live compatibly as pairs and that castration improves compatibility among males. Paired rabbits exhibit many postures and behaviors that are not possible in the small, solitary cages currently listed as minimum recommendations. PMID- 16363951 TI - Effects of inedible, manipulable objects on captive bears. AB - Bears in captivity often show abnormal behaviors such as stereotypies and periods of excessive inactivity. Most of the research aimed at reducing abnormal behaviors in bears has focused on feeding enrichment. However, the effects of feeding enrichment only seem to last as long as it takes a bear to consume the food. This study investigated the effects of inedible, manipulable objects on bears. Two polar bears (Ursus maritimus) received plastic floats. A sloth bear (Melursus ursinus) and a spectacled bear (Treniarctos ornatus) received plastic balls. During a 7-week period, polar bears nearly doubled their activity in the presence of toy floats, and the spectacled bear halved its pacing in the presence of toys. The sloth bear did not respond to the objects. These findings indicate that access to manipulable nonfood objects such as plastic toys can result in prolonged engagement in appetitive behaviors by polar bears. PMID- 16363952 TI - Housing density and growth in juvenile red-eared turtles. AB - Growth rates in juvenile turtles relate strongly to their health and welfare, both in the wild and in captivity. Newly hatched turtles in the wild are subject to such high levels of predation that, in some cases, as few as I % survive their first year of life. Juvenile turtles in captivity succumb more quickly to the effects of poor housing conditions and poor diets than do older adults. In both the wild and in captivity, turtles who grow quickly during their first year of life stand a better chance of survival to adulthood. Part of a long-term project to analyze the growth of juvenile turtles under laboratory conditions, this article seeks to develop a protocol for the successful maintenance of turtles in captivity by defining variables that affect health and survival. This article presents the findings of a study to determine whether variations in housing density affect the growth of juvenile red-eared turtles (Trachemys scripta elegans). PMID- 16363953 TI - Chimpanzees and retirement. AB - The subject of sanctuaries for chimpanzees has lately become the topic of a great deal of discussion (Brent, Butler, & Haberstroh, 1997; Committee on Long-Term Care of Chimpanzees, 1997; Dyke, Williams-Blangero, Mamelka, & Goodwin, 1995; Peterson & Goodall, 1993). In the United States, laboratories that use chimpanzees in research are facing a housing crisis. An increase in captive births caused by the initiation of the National Chimpanzee Breeding and Research Program in 1986 (Hobson, Graham, & Rowell, 1991), coupled with the diminished use of chimpanzees as experimental subjects, have led to a large population of chimpanzees considered to be surplus to demand (Blood, Wolfle, & Whitney, 1992). These chimpanzees, as well as an unknown number from the private sector, are candidates for what is currently being called retirement. PMID- 16363956 TI - Biology of animal stress: implications for animal well-being. PMID- 16363957 TI - Report on the 1998 ACLAM forum. PMID- 16363958 TI - Use of temporary and semipermanent enrichment objects by five chimpanzees. AB - At the Chimpanzee and Human Communication Institute, caregivers provide the 5 chimpanzees who reside there with many different forms of social, food, habitat, and object enrichment. In this study, we examined the chimpanzees' use of both semipermanent and temporary objects. Semipermanent objects included cargo nets, climbing structures, a treat mound, and other objects that were present at the chimpanzees' enclosure throughout the duration of this study. Each day, 50 temporary objects were placed in the chimpanzees' outdoor enclosure or indoor exercise rooms. Frequency of use was examined in 2 conditions: rotated and same. In the rotated condition, temporary objects were replaced with different temporary objects after 3 hr. In the same condition, temporary objects were presented for the entire day. Focal and scan sampling were used to record the chimpanzees' use of enrichment objects. Observers collected focal sample data to record the chimpanzees' initial reaction to objects when entering the indoor exercise rooms, outdoor enclosures, or both at 9:00 a.m. and 1.00 p.m. A total of 35 hr of focal data and 156 hr of scan data were collected over an 8-week period. Temporary object rotation increased the overall frequency of temporary object use both in the initial 15 min of focal sample data and during the following 6 hr of scan sample data for 4 of the chimpanzees. All of these chimpanzees used both semipermanent and temporary objects throughout the day. Each chimpanzee's pattern of use was unique. The results of this study emphasize the importance of temporary object rotation and presentation of both temporary and semipermanent objects to captive chimpanzee environments. PMID- 16363959 TI - Management of spent hens. AB - When hens kept for commercial egg production reach the end of their economically viable life span, they are usually transported to a processing plant for conversion into meat products. In this article, we review methods used in Canada and the United States for the catching and removing of these "spent" hens from the layer house and the subsequent loading of them onto vehicles for transport, unloading, and slaughter at the processing plant. We outline welfare concerns associated with these procedures with emphasis on bone fractures. We discuss the move toward on-farm euthanasia in North America and the humaneness of the various techniques under investigation. We conclude with recommendations aimed at increasing bone strength and improving the equipment and handling methods used in the movement and slaughter of spent hens. PMID- 16363960 TI - Segregated early weaning and welfare of piglets. AB - Segregated early weaning (SEW), introduced with the objectives of eradicating or controlling diseases and improving performance, is becoming increasingly popular among producers. The main features of SEW are weaning piglets early and moving the newly weaned piglets to a site isolated from the sows and other animals. In this article, we describe the key features of SEW and factors leading to its development. We review some of the main welfare issues involved with raising pigs in this way. We describe experiments that were crucial in the development of SEW and that showed positive effects on the health status and weight gain of piglets. We also review experiments that indicated welfare problems associated with this system. Specifically, we look at the effects of early weaning on the behavior and well-being of piglets. In addition, we look at other features of SEW, such as repeated cross-fostering and transport of young piglets. We provide directions for future research and offer recommendations for producers. PMID- 16363961 TI - Reasons for relinquishment of companion animals in U.S. animal shelters: selected health and personal issues. AB - In personal interviews, people surrendering their dogs and cats to 12 animal shelters in 4 regions of the country discussed their reasons for relinquishing their companion animals and answered questions about their own characteristics and those of their pet. The interviews identified 71 reasons for relinquishment. Personal issues lead the class of reasons for relinquishment of cats and ranked 3rd among those given for relinquishment of dogs. The top 3 health and personal issues cited for giving up cats were allergies of a family member to cats, owner's personal problems, and anew baby. For dogs, the top 3 reasons cited were lack of time for the dog, owner's personal problems, and allergies. Analysis of these health and personal issues suggests that education and counseling before and after acquisition of a pet, as well as the availability of temporary accommodations for pets during times of personal crisis, may reduce relinquishments. PMID- 16363962 TI - Animal abuse in Massachusetts: a summary of case reports at the MSPCA and attitudes of Massachusetts veterinarians. AB - This article reports information abstracted from 200 randomly sampled animal abuse complaints that the Massachusetts Society for the Prevention of Cruelty to Animals received in 1996, along with the results of 1 10 surveys that Massachusetts veterinarians completed concerning their experience with and attitudes and perceptions of animal mistreatment. In 1996, there were a total of 4,942 complaints of animal mistreatment, or 2.2 complaints per 1,000 households in Massachusetts. The majority of sampled complaints involved dogs (69.5%), cats (21.5%), or both. Almost all complaints involved husbandry-related neglect (62.0%), medical neglect (26.0%), or both. A violation of the law was observed by the investigating officer in 75 (37.5%) cases. The majority (78.9%) of veterinarians reported having observed at least 1 instance of animal abuse in their patients, although few encountered more than 5 cases (16.4%) during their years of practice. Almost all respondents (93.6%) agreed that veterinarians have an ethical responsibility to report suspicions of abuse, but a smaller number (44.5%) believed that this responsibility should be mandated. PMID- 16363964 TI - ECVAM workshop on alternatives to the use of animals in higher education. PMID- 16363965 TI - Population dynamics, overpopulation, and the welfare of companion animals: new insights on old and new data. AB - Americans profess a great love for their companion animals, and, indeed, their expenditures on food and other products for their dogs and cats would seem to confirm this. However, each year, many millions of dogs and cats are brought to animal shelters, where the majority are euthanized. Our inability to provide reasonably valid statistics related to this concern makes it difficult to offer a credible presentation on the need for a concerted effort to deal with the issue, design initiatives to ameliorate the problem, or evaluate progress and performance of these efforts. In this article we review some of the past efforts to document the scale of the "pet overpopulation" problem. We reexamine long-term shelter statistics from a single shelter system and present new data that reflect a recent cooperative effort to understand the origins and disposition of dogs and cats received by animal shelters. PMID- 16363966 TI - Human and animal factors related to relinquishment of dogs and cats in 12 selected animal shelters in the United States. AB - Since the 1940s, perceived companion animal overpopulation in the United States has been an important issue to the animal welfare community (Moulton, Wright, & Rinky, 1991). This surplus of animals has resulted in millions of dogs and cats being euthanized annually in animal shelters across the country. The nature and scope of this problem have been notoriously difficult to characterize. The number of animal shelters in the United Stares, the demographics of the population of animals passing through them, and the characteristics of per owners relinquishing animals are poorly understood. What portion of these animals are adopted or euthanized, why they are relinquished, and their source of acquisition are all questions for which there have been little data. Consequently, we are no closer to answering the fundamental question of how and why many animals are destroyed each year in shelters (Arkow, 1994). PMID- 16363967 TI - Cage toys reduce abnormal behavior in individually housed pigtail macaques. AB - As part of a behavioral intervention program that identifies and treats individual nonhuman primates exhibiting abnormal behavior, five individually housed pigtail macaques (Macaca nemestrina) were provided with multiple cage toys in an effort to reduce high levels of abnormal behavior. Ten 30-min observations of each subject were conducted during the baseline condition and again after novel toys were presented, both loose inside the cage and attached to the outside of the cage. The new toys were used during 27% of the observation time. Kong Toys were used most consistently by the macaques during the 5-week observation period. Significant decreases in abnormal behavior and cage-directed behavior, as well as significantly increased enrichment use, were evident after the toys were added. Several of the toys were destroyed quickly, and individual differences were evident in the levels of enrichment use and abnormal behavior. Providing multiple manipulable toys as enrichment for pigtail macaques was effective in reducing abnormal behavior and was an important part of an environmental enrichment program for monkeys who could not be housed socially. PMID- 16363968 TI - Welfare aspects of the transport by road of sheep and pigs. AB - That transport can be stressful to pigs and sheep has been inferred from behavioral and physiological measurements. Duration of journey is more likely to cause stress in pigs than in sheep, but loading and the start of travel are stressful to both. Vibration, related to vehicle design, and the jolting, shocks, and sudden impacts caused by road conditions and manner of driving, are probably of more importance than noise. Ventilation and stocking density can modulate the effects of ambient temperature, which may influence meat quality in pigs. Deprivation of food and water does not appear to stress sheep unduly, but this can become a serious welfare concern in the case of pigs. However, most journeys undertaken by pigs in the European Union are relatively short. In sheep, transport may follow soon after other stressful experiences such as weaning, shearing, handling, or marketing; the interaction of these factors in causing cumulative stress has not been studied. In pigs, fighting after the mixing of previously unacquainted animals is well known to be a welfare issue that can compromise meat quality. Considering the public interest in livestock transport and the large amounts of money involved, surprisingly little research has been done in the area, and more is urgently needed for legislation and welfare codes to be soundly based on scientific knowledge. PMID- 16363969 TI - Genetic engineering, welfare, and accountability. AB - Comments on the implications of genetic engineering for animal welfare. Welfare problems associated with techniques used to achieve genetic changes; Detrimental effects of genetic modifications to welfare; Modification of farm animals for biomedical products. Implications of genetic engineering for animal welfare are changing rapidly and need to be reviewed regularly. They include the welfare problems associated with techniques used to achieve genetic changes, which are similar to problems of other experimental approaches; these should be considered carefully, especially where techniques are used on a routine basis. When it comes to the genetic modifications themselves, some are detrimental to welfare, some are neutral, and some are beneficial; these results include direct effects of the intended change, side effects, and indirect effects. Currently, the two main applications are modification of farm animals for biomedical products--which appears to be largely neutral for welfare--and modification of mice as models for human disease, which results in suffering, often severe suffering. Beneficial applications are rare and still experimental or theoretical. The situation is similar with regard to the use of recombinant hormones and viruses; use of recombinant vaccines has potential for improving welfare, but may raise other ethical problems. Although few, if any, of these concerns are specific to genetic engineering, various factors combine to suggest that particular safeguards are needed in this field. These include the facts that changes can be produced rapidly and repeatedly, and that one of the driving forces behind genetic engineering is commercial exploitation of technology. In general, ethical evaluation still is done on a case-by-case basis, using the limited criteria seen as directly relevant to each case, rather than on a broader framework. There also is little public accountability, whereby the public can have confidence that such evaluation is being carried out properly. Calls for advisory "watchdog" committees to consider ethical questions on the use of animals are endorsed by this article. Furthermore, it is essential for public confidence in the safeguarding of animal welfare that the procedures of such committees should be well-publicized. PMID- 16363973 TI - Alternatives in monoclonal antibody production. PMID- 16363970 TI - Xenotransplantation: a problematic world behind a glamorous facade. AB - The discussion about the ethics of xenotransplantation seems to focus upon the benefits for individual patients and the potential risks for human society, in general, to contract a newly emerging retrovirus. In these risk-benefit considerations, the moral concern for the research animals involved appears to be absent. This is remarkable, because the presumed successful xenograft is not expected very soon. A lot of basic problems in pig and primate xenotransplantations still need solving. These new experiments in our own biomedical laboratories raise questions regarding animal welfare and ethical justification in the light of possible alternative strategies. In this article, I discuss some of the moral issues related to preclinical, fundamental xenotransplantation research. PMID- 16363974 TI - Thirty years after Brambell: whither animal welfare science? AB - It has been just over 30 years since the Brambell Committee issued its clarion call for research on the welfare of animals used in agriculture. What progress has farm animal welfare science made in those 30 years, and what challenges will it face in the next 30? In this article, I discuss the ways in which the Brambell report, with its emphasis on behavioral needs and suffering, both propelled and constrained the development of animal welfare science. The role that economic factors play and the ways in which they have imposed other kinds of constraints on scientists working with farm animals are also mentioned. Despite these constraints, animal welfare science has contributed to a number of significant improvements in the welfare of farm animals. There is, however, a growing sense that animal welfare science has reached an impasse and that ethical and scientific questions about animal welfare have become hopelessly entangled. In this context, I address what I view to be the principal challenge facing farm animal welfare science, namely to move the issue of beyond suffering to an evaluation of broader quality-of-life questions and their application to improvements in welfare. PMID- 16363975 TI - A new generation of antibodies. AB - Antibodies, proteins produced in animals that bind with high specificity and affinity to a seemingly limitless variety of biomolecules, have an essential role in clinical medicine and basic biological research. Methods to produce antibody fragments on the surface of bacterial viruses were surveyed for their ability to replace animal-dependent antibodies. The use of filamentous phage to display antibody fragments derived from semisynthetic antibody genes was found to produce proteins that bind to antigens with a variety, specificity, and affinity similar to those produced in animal systems. PMID- 16363976 TI - Computer simulation studies and the scientific method. AB - The scientific method is the formal procedure for all acceptable scientific endeavors. With this methodology, there is a continual interaction between theory, in the form of an hypothesis, and objective, experimental analysis. There is a new step in the scientific method that involves the use of computer models and simulation studies. When computer models are incorporated into hypothesis formulation, they can be used in simulation studies to test ideas before they are tried experimentally. An iterative feedback between these tests and current ideas allows for a preliminary refinement of hypotheses and development of more intelligent research protocols. In this way, computer simulation studies can serve as an intermediate step in the scientific method, reducing the number of animals used in biomedical experimentation. In this article we also explore other ways that computer simulation studies could limit the use of animals in biomedical research and education. PMID- 16363977 TI - Observations on assistance dog training and use. AB - Trained service dogs assist and also add pride, self-reliance, and personal satisfaction to an individual's daily life. However, well-bred, trained dogs are very expensive. To explore decreasing the cost of a service dog by increasing the number of dogs successfully completing training, the authors analyzed tasks that service dogs are expected to perform, measuring some of the inherent physical stresses the dogs encounter and considering training techniques to better prepare the dogs for more successful completion of tasks. Observing working dogs, the authors found that undue stress was placed on many of the dogs while performing tasks; that motivational gestures for performance were not fully extended; and that, many times, handlers did not instruct the dogs properly. The working dogs observed often were required to perform tasks that put them at risk of injury and performance failure. Important future considerations for training and handling working dogs will include proper training of handlers and a better understanding of the complexities of the instinctive and physical capabilities and limitation of dogs. PMID- 16363978 TI - History and future of animal welfare science. PMID- 16363983 TI - Editor's introduction to the Journal of Applied Animal Welfare Science. PMID- 16363984 TI - Destructible toys as enrichment for captive chimpanzees. AB - The use of destructible objects or toys as enrichment for nonhuman primates has had promising results in terms of increased use and positive behavioral effect. The purpose of this project was to determine the use and durability of a number of inexpensive, destructible toys provided one at a time or several at once. Nine singly caged chimpanzees were provided with 8 different toys made of plastic, vinyl, or cloth, and the frequency of use of the toys was determined during 15 min trials 2 times per day. A toy was removed when it was destroyed or when it was not contacted during 4 trials. The chimpanzees contacted the toys an average of 11 times per trial, and the use of the individual toys was significantly higher when provided 1 at a time rather than all at once. Use of the toys was fairly stable over time, and the toys remained in the cages an average of 3.2 days. The durability of the toys was related to the type of toy (e.g., more flexible cloth and vinyl toys lasted longer than rigid plastic toys). The destructible toys were used significantly more often than other permanent cage toys or televisions. Toy use was not related to age, level of abnormal behavior, or use of existing permanent toys or television. The implications of the results were related to the management of an environmental enrichment program and indicated that the provision of flexible, inexpensive toys 1 at a time can be an effective method of enrichment for captive chimpanzees. PMID- 16363985 TI - Unrelieved pain and distress in animals: an analysis of USDA data on experimental procedures. AB - Pain and distress are core issues in the field of animal experimentation and in the controversy that surrounds it. We sought to add to the empirical base of the literature on pain and distress by examining government data on experimental procedures that caused unrelieved pain and distress (UPAD) in animals. Of the species regulated by the U.S. Department of Agriculture (USDA), most of the approximately 100,000 animals subjected to UPAD during the year analyzed (1992) were guinea pigs and hamsters. Most of these animals were used in industry laboratories for various testing procedures, primarily vaccine potency testing. We discuss the limitations of the USDA data and recommend changes to the current reporting system. By identifying experimental procedures that cause UPAD in large numbers of USDA-regulated animals, the present analysis can be viewed as a means of identifying priorities for research and development of alternatives methods (replacements, reductions, and refinements). PMID- 16363986 TI - Development of a new production system for weaned and growing pigs. AB - The design of modem housing systems for farm animals calls for cooperation between scientists within different disciplines. This article describes the design of a new production system for weaned and growing pigs, as well as the analyses and working processes behind its development. The aim of the study was to develop an economically competitive system that also provided better welfare for the animals and a healthier environment for the workers. Analyses were carried out with emphasis on behavioral needs and building design and economy. The resulting system, the "safari system," consists of a number of straw flow pens of increasing size. Pigs are moved to larger and larger pens each week until slaughter. The main advantage is that pigs are not mixed during the growth period and that they are given more space, especially toward the end of the growth period when their space demand is high. Furthermore, the provision of straw for comfort and exploration and of wallow basins for cooling in the pens for the larger pigs were considered significant welfare improvements. An analysis revealed that the safari system would be economically competitive in comparison to traditional intensive systems. PMID- 16363987 TI - Abnormal behavior in caged birds kept as pets. AB - There are a limited number of studies dealing with abnormal behavior in caged birds kept as pets. However, these studies demonstrate the presence of abnormal behavior in both songbirds and parrots. Ethological studies on these birds, as well as studies on domestic and zoo birds, indicate that inappropriate rearing and housing conditions may lead to behavioral abnormalities. Together these data indicate that behavioral abnormalities occur among both wild-caught and domesticated pet birds. The severity and magnitude of these abnormalities is probably underestimated, and there is a need for systematic studies on the nature, origin, variability, species-specificity, and reversibility of behavioral problems in pet birds. Abnormal behavior in caged birds may to some extent be prevented and reduced by environmental enrichment. However, most enrichment studies are anecdotal and not based on a thorough analysis of the behavioral abnormalities, which may lead to measures resulting in a reduction of symptoms rather than the underlying causes. Although it is likely that several of these problems could be reduced by modifying rearing and housing conditions, the current insights into the causal mechanisms underlying abnormal behavior of domesticated and wild-caught pet birds are limited, as are the insights into the possibilities of preventing or curing abnormal behavior. PMID- 16363988 TI - On the psychological well-being of chimpanzees. PMID- 16363991 TI - The Second World Congress on Alternatives and Animal Use in the Life Sciences. PMID- 16363992 TI - Ca2+ release triggered by NAADP in hepatocyte microsomes. AB - NAADP (nicotinic acid-adenine dinucleotide phosphate) is fast emerging as a new intracellular Ca2+-mobilizing messenger. NAADP induces Ca2+ release by a mechanism that is distinct from IP3 (inositol 1,4,5-trisphosphate)- and cADPR (cADP-ribose)-induced Ca2+ release. In the present study, we demonstrated that micromolar concentrations of NAADP trigger Ca2+ release from rat hepatocyte microsomes. Cross-desensitization to IP3 and cADPR by NAADP did not occur in liver microsomes. We report that non-activating concentrations of NAADP can fully inactivate the NAADP-sensitive Ca2+-release mechanism in hepatocyte microsomes. The ability of thapsigargin to block the NAADP-sensitive Ca2+ release is not observed in sea-urchin eggs or in intact mammalian cells. In contrast with the Ca2+ release induced by IP3 and cADPR, the Ca2+ release induced by NAADP was completely independent of the free extravesicular Ca2+ concentration and pH (in the range 6.4-7.8). The NAADP-elicited Ca2+ release cannot be blocked by the inhibitors of the IP3 receptors and the ryanodine receptor. On the other hand, verapamil and diltiazem do inhibit the NAADP- (but not IP3- or cADPR-) induced Ca2+ release. PMID- 16363993 TI - Misincorporation of free m-tyrosine into cellular proteins: a potential cytotoxic mechanism for oxidized amino acids. AB - In vitro studies demonstrate that the hydroxyl radical converts L-phenylalanine into m-tyrosine, an unnatural isomer of L-tyrosine. Quantification of m-tyrosine has been widely used as an index of oxidative damage in tissue proteins. However, the possibility that m-tyrosine might be generated oxidatively from free L phenylalanine that could subsequently be incorporated into proteins as an L tyrosine analogue has received little attention. In the present study, we demonstrate that free m-tyrosine is toxic to cultured CHO (Chinese-hamster ovary) cells. We readily detected radiolabelled material in proteins isolated from CHO cells that had been incubated with m-[14C]tyrosine, suggesting that the oxygenated amino acid was taken up and incorporated into cellular proteins. m Tyrosine was detected by co-elution with authentic material on HPLC and by tandem mass spectrometric analysis in acid hydrolysates of proteins isolated from CHO cells exposed to m-tyrosine, indicating that free m-tyrosine was incorporated intact rather than being metabolized to other products that were subsequently incorporated into proteins. Incorporation of m-tyrosine into cellular proteins was sensitive to inhibition by cycloheximide, suggesting that protein synthesis was involved. Protein synthesis using a cell-free transcription/translation system showed that m-tyrosine was incorporated into proteins in vitro by a mechanism that may involve L-phenylalanine-tRNA synthetase. Collectively, these observations indicate that m-tyrosine is toxic to cells by a pathway that may involve incorporation of the oxidized amino acid into proteins. Thus misincorporation of free oxidized amino acids during protein synthesis may represent an alternative mechanism for oxidative stress and tissue injury during aging and disease. PMID- 16363994 TI - Enhanced levels of Pis1p (phosphatidylinositol synthase) improve the growth of Saccharomyces cerevisiae cells deficient in Rsp5 ubiquitin ligase. AB - The Rsp5 ubiquitin ligase plays a role in many cellular processes including the biosynthesis of unsaturated fatty acids. The PIS1 (phosphatidylinositol synthase gene) encoding the enzyme Pis1p which catalyses the synthesis of phosphatidylinositol from CDP-diacyglycerol and inositol, was isolated in a screen for multicopy suppressors of the rsp5 temperature sensitivity phenotype. Suppression was allele non-specific. Interestingly, expression of PIS1 was 2-fold higher in the rsp5 mutant than in wild-type yeast, whereas the introduction of PIS1 in a multicopy plasmid increased the level of Pis1p 6-fold in both backgrounds. We demonstrate concomitantly that the expression of INO1 (inositol phosphate synthase gene) was also elevated approx. 2-fold in the rsp5 mutant as compared with the wild-type, and that inositol added to the medium improved growth of rsp5 mutants at a restrictive temperature. These results suggest that enhanced phosphatidylinositol synthesis may account for PIS1 suppression of rsp5 defects. Analysis of lipid extracts revealed the accumulation of saturated fatty acids in the rsp5 mutant, as a consequence of the prevention of unsaturated fatty acid synthesis. Overexpression of PIS1 did not correct the cellular fatty acid content; however, saturated fatty acids (C(16:0)) accumulated preferentially in phosphatidylinositol, and (wild-type)-like fatty acid composition in phosphatidylethanolamine was restored. PMID- 16363997 TI - Explorative statistical analysis of planar point processes in microscopy. AB - Basic methods of explorative statistical analysis for stationary and isotropic planar point processes are briefly and informally reviewed. At the explorative level, planar point patterns may be characterized in terms of the intensity, the K-function and the pair correlation function. These second-order functions enable one to classify a given point process as completely random, clustering or repulsive. The repulsive behaviour may be quantified by an estimate of the hard core distance. In the exploratory approach, the statistics are essentially free from model assumptions. Second-order spatial functions have been estimated to characterize genuine planar point processes in the macroscopic domain, for example in forestry, geography and epidemiology. For light microscopy and transmission electron microscopy, two situations are distinguished, which may be summarized as the genuine planar case and the stereological case. In the genuine planar case, a direct interpretation of the results of spatial statistics is feasible. Here, monolayers in cell culture, intramembranous particles on freeze fracture specimens and amacrine cells of the retina are mentioned as examples. In the stereological case, point patterns are generated by sections through 3D structures. Here the observed point patterns may arise as the centres of sectional profiles of particles, or as centres of sectional profiles of spatial fibre processes. In both situations, exploratory spatial point process statistics allow a quantitative characterization of sectional images for the purposes of group comparisons and classification. Moreover, for spatial fibre processes it has recently been shown that the observed pair correlation function of the centres of the fibre profiles is an estimate of the reduced pair correlation function of the fibre process in 3D. Hence for fibre processes a stereological interpretation of point process statistics obtained from sections is an additional option. PMID- 16363995 TI - Cocaine-induced alterations in nucleus accumbens ionotropic glutamate receptor subunits in human and non-human primates. AB - Chronic cocaine and withdrawal induce significant alterations in nucleus accumbens (NAc) glutamatergic function in humans and rodent models of cocaine addiction. Dysregulation of glutamatergic function of the prefrontal cortical-NAc pathway has been proposed as a critical substrate for unmanageable drug seeking. Previously, we demonstrated significant up-regulation of NMDA, (+/-)-alpha-amino 3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptor subunit mRNAs and protein levels in the ventral tegmental area (VTA), but not the substantia nigra, of cocaine overdose victims (COD). The present study was undertaken to examine the extent of altered ionotropic glutamate receptor (iGluR) subunit expression in the NAc and the putamen in cocaine overdose victims. Results revealed statistically significant increases in the NAc, but not in the putamen, of NMDA receptor subunit (NR)1 and glutamate receptor subunit (GluR)2/3 wit trends in GluR1 and GluR5 in COD. These results extend our previous finding and indicate pathway-specific alterations in iGluRs in COD. In order to determine that changes were related to cocaine intake and not to other factors in the COD victims, we examined the effects of cocaine intravenous self-administration in rhesus monkeys for 18 months (unit dose of 0.1 mg/kg/injection and daily drug intake of 0.5 mg/kg/session). Total drug intake for the group of four monkeys was 37.9 +/- 4.6 mg/kg. Statistically significant elevations were observed for NR1, GluR1, GluR2/3 and GluR5 (p < 0.05) and a trend towards increased NR1 phosphorylated at serine 896 (p = 0.07) in the NAc but not putamen of monkeys self-administering cocaine compared with controls. These results extend previous results by demonstrating an up-regulation of NR1, GluR2/3 and GluR5 in the NAc and suggest these alterations are pathway specific. Furthermore, these changes may mediate persistent drug intake and craving in the human cocaine abuser. PMID- 16363999 TI - Tracking quasi-stationary flow of weak fluorescent signals by adaptive multi frame correlation. AB - We have developed a novel cross-correlation technique to probe quasi-stationary flow of fluorescent signals in live cells at a spatial resolution that is close to single particle tracking. By correlating image blocks between pairs of consecutive frames and integrating their correlation scores over multiple frame pairs, uncertainty in identifying a globally significant maximum in the correlation score function has been greatly reduced as compared with conventional correlation-based tracking using the signal of only two consecutive frames. This approach proves robust and very effective in analysing images with a weak, noise perturbed signal contrast where texture characteristics cannot be matched between only a pair of frames. It can also be applied to images that lack prominent features that could be utilized for particle tracking or feature-based template matching. Furthermore, owing to the integration of correlation scores over multiple frames, the method can handle signals with substantial frame-to-frame intensity variation where conventional correlation-based tracking fails. We tested the performance of the method by tracking polymer flow in actin and microtubule cytoskeleton structures labelled at various fluorophore densities providing imagery with a broad range of signal modulation and noise. In applications to fluorescent speckle microscopy (FSM), where the fluorophore density is sufficiently low to reveal patterns of discrete fluorescent marks referred to as speckles, we combined the multi-frame correlation approach proposed above with particle tracking. This hybrid approach allowed us to follow single speckles robustly in areas of high speckle density and fast flow, where previously published FSM analysis methods were unsuccessful. Thus, we can now probe cytoskeleton polymer dynamics in living cells at an entirely new level of complexity and with unprecedented detail. PMID- 16364000 TI - Two-photon fluorescence surface wave microscopy. AB - This paper demonstrates the principle of two-photon surface wave microscopy with a view to applications on biological samples. We describe a modified scanning optical microscope, which uses specially prepared coverslips. These coverslips are designed to support the propagation of surface waves capable of large field enhancements. We also discuss the beam conditioning necessary to ensure efficient use of the available illumination. Two-photon surface wave fluorescent excitation is demonstrated on fluorescent nanospheres, demonstrating a point spread function width of approximately 220 nm at an illumination wavelength of 925 nm. The potential of non-linear surface wave excitation for both fluorescence and harmonic imaging microscopy is discussed. PMID- 16364001 TI - Studies on depth-of-field effects in microscopy supported by numerical simulations. AB - Micrographs are two-dimensional (2D) representations of three-dimensional (3D) objects. When the depth-of-field of a micrograph is comparable with or larger than the characteristic dimension of objects within the micrograph, measured 2D parameters (e.g. particle number density, surface area of particles, fraction of open space) require stereological correction to determine the correct 3D values. Here, we develop a stereological theory using a differential approach to relate the 3D volume fraction and specific surface area to the 2D projected area and perimeter fractions, accounting for the influence of depth-of-field. The stereological theory is appropriate for random isotropic arrangements of non interpenetrating particles and is valid for convex geometries (e.g. spheres, spheroids, cylinders). These geometrical assumptions allow the stereological formulae to be expressed as a set of algebraic equations incorporating a single parameter to describe particle shape that is tightly bounded between 1.5pi and 2pi. The stereological theory may also be applied to arrangements of interpenetrating convex particles, and for this case, the resulting stereological formulae become identical to the formulae previously presented by Miles. To test the accuracy of the stereological theory, random computational arrangements of non-interpenetrating and interpenetrating spheres or cylinders are analysed, and the projected area and perimeter fractions are numerically determined as a function of depth-of-field. The computational results show very good agreement with the theoretical predictions over a broad range of depth-of-field, volume fraction and particle geometry for both non-interpenetrating and interpenetrating particles, demonstrating the overall accuracy of the stereological theory. Applications of the stereological theory towards analysis of biological tissues and extracellular matrix are discussed. PMID- 16364002 TI - Segmentation of intestinal gland images with iterative region growing. AB - A region growing algorithm for segmentation of human intestinal gland images is presented. The initial seeding regions are identified based on the large vacant regions (lumen) inside the intestinal glands by fitting with a very large moving window. The seeding regions are then expanded by repetitive application of a morphological dilate operation with a much smaller round window structure set. False gland regions (nongland regions initially misclassified as gland regions) are removed based on either their excessive ages of active growth or inadequate thickness of dams formed by the strings of goblet cell nuclei sitting immediately outside the grown regions. The goblet cell nuclei are then identified and retained in the image. The gland contours are detected by applying a large moving round window fitting to the enormous empty exterior of the goblet cell nucleus chains in the image. The assumptions based on real intestinal gland images include the closed chain structured goblet cell nuclei that sit side-by-side with only small gaps between the neighbouring nuclei and that the lumens enclosed by the goblet cell nucleus chains are most vacant with only occasional run-away nuclei. The method performs well for most normal and abnormal intestinal gland images although it is less applicable to cancer cases. The experimental results show that the segmentations of the real microscopic intestinal gland images are satisfactorily accurate based on the visual evaluations. PMID- 16364006 TI - Antigen presentation by human uterine epithelial cells to autologous T cells. AB - PROBLEM: Epithelial cells, as sentinels of immune protection in the endometrium, use innate immune mechanisms to protect against infection from pathogenic microbes. Our goal in this study was to assess the ability of human uterine epithelial cells to present antigen to cells of the adaptive immune system. METHOD OF STUDY: Highly purified preparations of uterine epithelial cells from 11 patients were assessed for their ability to present tetanus toxoid (TT) to autologous T cells. Leukocyte contamination in the epithelial cell preparations was numerically and functionally determined. Using confocal microscopy, epithelial cells were tested for the expression of CD40 and CD1d. RESULTS: Purified preparations of endometrial epithelial cells isolated from every patient presented TT recall antigen to autologous T cells. Leukocyte contamination of epithelial cell preparations was insignificant. Uterine epithelial cells express CD40 and CD1d. CONCLUSION: Antigen presentation is an additional aspect of uterine epithelial cell function in maintaining women's health. PMID- 16364007 TI - Alterations in syncytiotrophoblast cytokine expression following treatment with lipopolysaccharide. AB - PROBLEM: The placental syncytium is a differentiated cell type on the surface of the villus that has the potential to release cytokines directly to maternal blood. Responsiveness of this cell type to inflammatory compounds remains largely unelucidated. METHOD OF STUDY: Response to a pro-inflammatory (lipopolysaccharide, LPS) and an anti-inflammatory (dexamethasone, DEX) compound was studied in primary cultures of syncytiotrophoblasts (SCTs). Cells were incubated with and without LPS and DEX. Cytokine levels in conditioned media were determined by enzyme-linked immunosorbent assay and proteome arrays. RESULTS: LPS treatment induced a fourfold increase in interleukin-8 (IL-8) levels in SCTs. LPS enhanced the expression of both pro- and anti-inflammatory cytokines in SCTs. DEX treatment reduced IL-8 levels in control and LPS-treated cultures by 70-90%. CONCLUSION: Cytokine expression in SCTs was enhanced by LPS treatment and this effect was suppressed by glucocorticoid treatment. This suggests that inflammatory compounds may alter cytokine expression in the syncytium throughout gestation. PMID- 16364008 TI - Evidence for interleukin-10-mediated inhibition of cyclo- oxygenase-2 expression and prostaglandin production in preterm human placenta. AB - PROBLEM: Interleukin-10 (IL-10) is thought to be a key cytokine for the maintenance of pregnancy. Here we examined the expression profiles of IL-10 and cyclo-oxygenase-2 (COX-2), and the effect of IL-10 on COX-2 expression and prostaglandin release in the human placenta from preterm labor deliveries associated with chorioamnionitis. METHOD OF STUDY: Placental tissues from preterm labor and term labor deliveries were processed for ex vivo placental explant culture system. IL-10 expression was assessed by enzyme-linked immunosorbent assay (ELISA) and immunohistochemical (IHC) analysis. COX-2 expression was evaluated by IHC, Western blotting and reverse transcriptase-polymerase chain reaction. Prostaglandin E2 (PGE2) release was measured by ELISA. RESULTS: IL-10 was significantly reduced in chorioamnionitis-associated preterm labor as well as in term labor placental tissues compared with second trimester normal pregnancy samples obtained from elective terminations. Similar results were obtained with freshly isolated cytotrophoblasts from these deliveries. As expected, COX-2 mRNA was detected at significant levels in tissues from term and preterm labor deliveries compared with no labor term deliveries. Importantly, IL-10 inhibited COX-2 expression in cultured placental explants from preterm labor deliveries, but not from term labor samples. Inhibition of COX-2 expression coincided with reduced PGE2 release. CONCLUSION: These results demonstrate the importance of IL 10 in countering inflammation associated with preterm labor, and suggest that term and preterm parturition may, in part, represent different conditions. PMID- 16364009 TI - Comparison of T cell subsets between somatic cloned and normal cow. AB - PROBLEM: Somatic cloning technology is beneficial for genetically producing excellent animals. However, many developmental problems of somatically cloned animals have been described. Some of them may cause disorders of the immune system, resulting in the fluctuation of the proportion of white blood cells (WBC), different from that of normal animals in peripheral blood. METHOD OF STUDY: In Holstein- cloned and normal cows, the fluctuation of granulocytes, monocytes, B cells and T cells, and further T cell subsets (CD4+, CD8+, gammadelta, CD8+gammadelta and WC1+gammadelta T cell) in peripheral blood were analyzed in early lactation stage (ELS) and mid to late lactation stage (MLS) by flow cytometry using specific monoclonal antibodies for cell surface markers. RESULTS: In both ELS and MLS, there were no significant differences in the proportions of granulocytes, monocytes, B cells and T cells between cloned and normal cows. In T cell subsets, gammadelta and WC1+gammadelta T cells in cloned cows were significantly less frequent than in normal cows in ELS. The decreased proportions of gammadelta and WC1+gammadelta T cells recovered to the level of normal cows in MLS. CONCLUSIONS: The population of granulocytes, monocytes, B cells and T cells, and T cell subsets except for gammadelta and WC1+gammadelta T cells in cloned cows fluctuated in a manner similar to those of normal cows during lactation. In ELS, the proportions of gammadelta and WC1+gammadelta T cells temporarily declined in cloned cows, suggesting that cloned cows may fall into an immunosuppressive state in ELS. PMID- 16364010 TI - Maternal immunopotentiation affects caspase activation and NF-kappaB DNA-binding activity in embryos responding to an embryopathic stress. AB - PROBLEM: Increased embryonic resistance to teratogenic stresses as a result of maternal immunopotentiation is associated with a decrease in the intensity of teratogen-induced apoptosis in target embryonic structures. These findings suggest that this effect of maternal immunopotentiation might be realized through modification of the expression of molecules regulating the teratogen-induced apoptotic process. To examine this possibility, we evaluated caspases 3, 8 and 9 activation as well as nuclear factor (NF)-kappaB DNA-binding activity in the embryos of immunopotentiated mice exposed to cyclophosphamide (CP). METHODS OF STUDY: The rate of resorptions and the proportion of malformed fetuses in CP treated mice were recorded on day 19 of pregnancy. Activity of caspases was tested in cytoplasmic extracts collected from the embryonic brain 24 hr after CP treatment using appropriate fluorometric kits, whereas NF-kappaB DNA-binding activity was evaluated in nuclear extracts using the electrophoretic mobility shift assay. RESULTS: As in our previous studies, immunopotentiated CP-treated females exhibited a lower rate of resorptions or fetuses with open eyes than their non-immunopotentiated counterparts. In parallel, we observed that maternal immunopotentiation normalized the CP-induced activation of the tested caspases as well as the CP-induced suppression of NF-kappaB DNA-binding activity. CONCLUSIONS: As caspases act as inducers of apoptosis, and NF-kappaB acts in CP treated embryos as an apoptosis suppressor, the above results suggest that maternal immunopotentiation might affect embryonic sensitivity to embryopathic stresses via NF-kappaB- and caspases-associated pathways. PMID- 16364011 TI - Aspirin desensitization in the treatment of antiphospholipid syndrome during pregnancy in ASA-sensitive patients. AB - PROBLEM: Antiphospholipid syndrome (APS) is associated with thrombosis and poor pregnancy outcome in the presence of antiphospholipid antibodies (aPL). Patients with aPL have a high risk of foetal loss. However, with low-dose aspirin (acetylsalicylic acid; ASA) in combination with subcutaneous heparin, the chances of full-term delivery increase. Nevertheless, ASA treatment is avoided in pregnant, ASA-sensitive women with APS. METHODS: Rapid oral challenge desensitization to ASA was performed in four pregnant women with a history of APS and aspirin sensitivity. In three patients, desensitization was performed during pregnancy and before the next pregnancy in the fourth. Desensitization was carried out in the ICU using increasing doses of aspirin (0.1-125 mg) over a 24 hr period. RESULTS: Successful ASA desensitization was achieved in all the patients. No severe side effects occurred during the desensitization test. Only one patient required a small oral dose of antihistamines. CONCLUSIONS: Aspirin desensitization may be a safe alternative even during pregnancy if carefully monitored and permit patients with APS to receive treatment with ASA. This would constitute a new indication in pregnant women with APS and ASA sensitivity. PMID- 16364012 TI - The aryl hydrocarbon receptor nuclear translocator gene polymorphism in patients with recurrent miscarriage. AB - OBJECTIVE: The human aryl hydrocarbon receptor nuclear translocator gene (ARNT) is crucial for embryonic development. Knockout of ARNT is embryonic lethal. We thus hypothesized that some cases of recurrent miscarriage (RM) may be due to a polymorphism in the ARNT gene. METHODS: Polymerase chain reaction was used to compare the gene frequencies of a polymorphism in codon 511 of the ARNT gene in 170 women with idiopathic RM and 105 controls. RESULTS: Using the Hardy-Weinberg equilibrium calculation, the predicted ARNT genotype frequencies for the N511/N511, N511/D511, and D511/D511 genotypes were 0, 2, and 167 respectively. The observed frequencies were 0, 2, and 168 (NS). The N/511/D511 genotype was detected in 1.2% of cases and 2.8% of controls, and the D511/D511genotype was detected in 98.8% of cases and 97.2% of controls (NS). CONCLUSIONS: In our cohort of patients, the polymorphism in codon 511 of the ARNT gene is not associated with RM. PMID- 16364013 TI - Testicular isoform of angiotensin I-converting enzyme (ACE, CD143) on the surface of human spermatozoa: revelation and quantification using monoclonal antibodies. AB - PROBLEM: The elucidation of the role of angiotensin-converting enzyme (ACE, CD143) in the male fertility has been hampered by the absence of highly specific antibodies to the native testicular isoform (tACE). The quantification of tACE expression on human-ejaculated spermatozoa was performed using a novel panel of monoclonal antibodies (mAbs). METHOD OF STUDY: The expression of tACE on the surface of live and fixed human spermatozoa was analyzed by flow cytometry and immunocytochemistry using new mAbs to human tACE. RESULTS: Monoclonal antibodies 1E10 and 4E3 similarly revealed tACE on the surface of live and fixed spermatozoa. The high percentage of tACE-positive spermatozoa (median 81%) was revealed in the swim-up fraction of sperm. Antibody-induced tACE shedding occurs preferentially from live sperm with defective function and/or morphology. Testicular ACE is located on the plasma membrane of the post-acrosomal region, the neck and midpiece of normal spermatozoa, but showed a variable distribution on the defective cells. CONCLUSIONS: The new mAbs recognizing the C-terminal domain of human ACE are useful tools for quantification of tACE expression on human live and fixed spermatozoa and further adequate analysis of the tACE role in reproduction. PMID- 16364014 TI - Granulocyte-macrophage colony-stimulating factor levels in amniotic fluid before the onset of labor and during labor do not differ in normal pregnancies. AB - PROBLEM: Granulocyte-macrophage colony-stimulating factor (GM-CSF) at the implantation site may regulate invasion and differentiation of placental trophoblast. We evaluated whether GM-CSF levels in amniotic fluid during labor contributing to subsequent delivery differed from those before the onset of labor in normal pregnancies. METHOD OF STUDY: This study enrolled 36 Japanese women experiencing normal pregnancies with single fetuses who had no infection. Of these pregnancies, 18 were women during labor that led to subsequent term delivery (labors). The other 18 were women without labor underwent cesarean section (controls). These two groups (18 labors and 18 controls) were compared. The average gestational age at entry was 38-39 weeks of gestation. The women's ages and gestational ages did not differ significantly between the two groups. Amniotic fluid was collected and the GM-CSF levels were compared between two groups. The GM-CSF level was determined by the enzyme-linked immunosorbent assay method. RESULTS: There was no significant increase in GM-CSF levels in amniotic fluid during labor compared with that before the onset of labor. CONCLUSIONS: The GM-CSF in amniotic fluid may not promote the onset of labor at term and/or term labor contributing to subsequent delivery may not induce the production and secretion of GM-CSF into amniotic cavity. PMID- 16364015 TI - The effect of serum concentration of leukaemia inhibitory factor on in vitro fertilization treatment outcome. AB - PROBLEM: To evaluate the association of peripheral leukaemia inhibitory factor (LIF) levels on implantation and miscarriage rates after in vitro fertilization (IVF) treatment. METHODS: Prospective observational study of 120 randomly selected women who underwent IVF treatment. The concentration of LIF in serum was determined by enzyme-linked immunosorbent assay. RESULTS: There was no significant differences with regard to the systemic mean LIF concentration between the pregnant (42 patients, LIF: 11.55 pg/mL +/- 5.3 S.D.) and non pregnant (66 patients, LIF: 13.47 pg/mL +/- 5.1 S.D.) women after IVF treatment. Likewise, for those women who have positive pregnancy after IVF treatment, the systemic mean LIF levels were not significantly different between women who have an ongoing pregnancy (34 ongoing pregnancy, LIF: 11.26 pg/mL +/- 5.2 S.D.) and those who had miscarriage (eight miscarriage, LIF: 12.78 pg/mL +/- 5.6 S.D.). CONCLUSION: The systemic levels of LIF concentration have no association with implantation rate or miscarriage rate in women undergoing IVF treatment. Measuring serum LIF concentration prior to embryo transfer in IVF treatment has no predictive value of implantation rate or miscarriage rate. PMID- 16364016 TI - Eimeriosis in cattle: current understanding. AB - This report addresses various aspects of the protozoan parasite Eimeria which contribute to their increasing recognition as important protozoal pathogens in cattle. Among others, questions of parasite biology and epidemiology, its impact on host physiology, and control strategies are dealt with. The tenacious oocysts are found ubiquitously in the environment making an infection of calves and young cattle, the most susceptible age group, almost inevitable. Further development, comprising of asexual multiplication, the merogony, and a subsequent sexual stage, the gamogony, takes place within cells of the small and large intestines, after which numerous unsporulated oocysts are formed and shed with the faeces. Of the more than a dozen species, Eimeria bovis and Eimeria zuernii only are made responsible for severe clinical disease characterized by haemorrhagic diarrhoea with sometimes fatal outcome. To a lesser extent, Eimeria alabamensis also can cause clinical disease. Because of the damage inflicted on the intestinal tissue, the digestive process and overall homeostasis can become severely affected, even with absence of clinical disease, with adverse effects on animal welfare and performance. The consequent economic losses for the cattle industry are thus substantial. Active (species specific) immunity, both humoral and cellular, develops rapidly after first antigen contact, its intensity being dependent on the number of oocysts ingested. However, no absolute protection is achieved and even older animals can excrete oocysts, contributing to a state of endemic stability. For efficient control, exact diagnosis of the Eimeria species involved and the evaluation of animal management and husbandry practices are of utmost importance. Mixed infections are the rule and only an occurrence of pathogenic species, together with clinical symptoms, justifies the assumption of a coccidiosis. Proper hygiene regime and ensuring unfavourable conditions for oocyst survival in the environment (cool and dry, disinfection procedures) will help to reduce infection pressure on the herd. Of the various pharmacological compounds used to treat coccidiosis, those which act against the late developmental stages, administered during prepatency (metaphylactically rather than therapeutically), are currently considered most effective. PMID- 16364017 TI - Virus spread pattern within infected chicken farms using regression model: the 2003-2004 HPAI epidemic in the Republic of Korea. AB - During the 2003-2004 epidemic in Korea, the infection time and within-farm spread pattern of virus were analysed for the highly pathogenic avian influenza (HPAI) outbreak on chicken farms using regression models based on epidemiological data. Mortality observed on a given day had a positive linear association with time after initial infection. HPAI spread more rapidly on farms managed by employees and on farms with larger numbers of chicken houses in use. The disease spread more rapidly among layer chickens than among broilers. Using statistical model, we found that farmers recognize the abnormally high mortality resulting from HPAI approximately 5 days after infection. Without any intervention, entire flocks would die within 12 days of introduction of the HPAI virus to the infected farm. PMID- 16364018 TI - Long-term studies on maternal immunity for Aujeszky's disease and classical swine fever in wild boar piglets. AB - The aim of the studies was to fathom the duration and the role of maternal immunity for Aujeszky's disease (AD) and classical swine fever (CSF) in wild boar offspring. In one experiment, two wild boar sows were infected with a low pathogenic pseudorabies virus (PRV) in 1999. A total of 51 offspring was born between 1999 and 2002 and was monitored for PRV maternal antibodies. In a second experiment, the maternal immunity for CSF was analysed. Therefore, a sow was orally vaccinated against CSF using vaccine baits containing the live-attenuated C-strain vaccine. The vaccination took place in January 1999. The sow gave birth to four piglets in 2001 and to two piglets in 2002. With respect to maternal immunity for AD, some piglets reacted positive in the ELISA up to 27-week post partum while in the neutralization test antibodies were detected up to 15-week post-partum. The calculated half-life of neutralizing antibodies was 21 days. Regarding CSF, the neutralization titres of maternal antibodies dropped continuously reaching values of < or =10 ND50 20-week post-partum. After the 12th week post-partum, most of the sera reacted negative in the ELISA. However, after the third month, low levels of neutralization titres were still detectable. The results are discussed with respect to the epidemiology and control of both diseases in wild boar populations. PMID- 16364019 TI - Comparative study of the effects of several chemical and physical treatments on the activity of protease resistance and infectivity of scrapie strain 263K. AB - To study the influences of chemical and physical factors on the protease resistant activity in vitro and the infectivity in vivo of scrapie strain 263K, PrPSc from the hamsters infected intracerebrally with scrapie strain 263K were treated with several commonly used disinfection methods, including sodium hydroxide (NaOH), sodium hypochlorite (NaOCl), heating or autoclaving at 80, 100, 121 and 134 degrees C in the solutions with or without 3% sodium dodecyl sulphate (SDS). The protease resistance of PrPSc was analysed by a proteinase K (PK) digesting Western blot and the infectivity of PrPSc was analysed by intracerebral (i.c.) inoculation into experimental hamsters. The results showed that PrPSc signals were removed in the preparations treated with NaOH higher than 0.05 mol/l, NaOCl higher than 0.1%, autoclaved over 121 degrees C, or heated over 80 degrees C in the presence of 3% SDS. Animal challenges revealed that mixing with 2 mol/l NaOH or 2% NaOCl, autoclaving at 134 degrees C, as well as heating at 100 degrees C or autoclaving at 121 degrees C in the solutions with 3% SDS completely blocks the transmission of scrapie 263K in this experimental situation. It is obvious that the removal of PK resistance of PrPSc happened at relatively lower concentration chemicals or lower temperature, while elimination of the infectivity needs more vigorous conditions. Our data provide the useful evidences for several commonly used methods to inactivate TSEs agent and suggest that it is inappropriate to use PrPSc as a surrogate for TSEs infectivity in inactivation experiments. PMID- 16364020 TI - Seroprevalence of brucellosis, tularemia, and yersiniosis in wild boars (Sus scrofa) from north-eastern Germany. AB - Brucellosis and tularemia are classical zoonotic diseases transmitted from an animal reservoir to humans. Both, wildlife and domestic animals, contribute to the spreading of these zoonoses. The surveillance of the animal health status is strictly regulated for domestic animals, whereas systematic disease monitoring in wildlife does not exist. The aim of the present study was to provide data on the prevalence of anti-Brucella, anti-Francisella and anti-Yersinia antibodies in wild boars from North-Eastern Germany to assess public health risks. A total of 763 sera of wild boars from Mecklenburg-Western Pomerania hunted in 1995/1996 were tested using a commercially available Brucella suis ELISA, an in-house lipopolysaccharide (LPS)-based Francisella ELISA, and commercially available Western blot kits for the detection of anti-Francisella and anti-Yersinia antibodies. The Yersinia enterocolitica O:9 LPS is able to induce serological cross-reactions indistinguishable from brucellosis due to a similar immunodominant epitope in the Brucella O-polysaccharide. The Yersinia Western blot assay was, therefore, based on five recombinant Yersinia outer proteins which have been proved to be specific for the serodiagnosis of yersiniosis. Anti Brucella, anti-Francisella and anti-Yersinia antibodies were detected in 22.0%, 3.1%, and 62.6% of the wild boars, respectively. The high seroprevalence of tularemia and brucellosis in wild boars indicates that natural foci of these zoonoses are present in wildlife in Germany. However, the impact of transmission of zoonotic pathogens from wildlife to livestock is unknown. Only careful and systematic monitoring will help to prevent the (re)emergence of these zoonotic diseases in domestic animals and consequently human infection. PMID- 16364021 TI - Identification of an aadA2 gene cassette from Salmonella enterica subsp. enterica Serovar derby. AB - During a study on Salmonella enterica subsp. enterica serovar Derby from slaughter-age pigs in Brazil, two epidemiologically unrelated multi-resistant S. Derby isolates were found to carry a class 1 integron with a single gene cassette. Sequence analysis confirmed that this gene cassette harboured an aadA2 gene. The aadA2 gene codes for an aminoglycoside adenyltransferase, which mediates resistance to the aminoglycoside streptomycin and the aminocyclitol spectinomycin. Although aadA2 gene cassettes are widely distributed among Salmonella, database searches identified an AadA2 protein indistinguishable from that of S. Derby only in single isolates of S. enterica subsp. enterica Enteritidis from France and S. enterica subsp. enterica Typhimurium from Japan. Structural analysis of the 59-base element revealed at least one base pair difference between the 59-base element of the aadA2 cassette from S. Derby and any of the 59-base elements deposited in the databases. PMID- 16364022 TI - Emergence of fluoroquinolone resistance in Campylobacter jejuni in chickens exposed to enrofloxacin treatment at the inherent dosage licensed in Japan. AB - The objective of the present study was to determine whether selection of fluoroquinolone resistance could be easily induced in Campylobacter jejuni colonized chickens by treatment with enrofloxacin of representative fluoroquinolones at the inherent dosage licensed in Japan (50 ppm in drinking water for 3 days). In the case of isolates from chickens of study 1, an increase in the population of susceptible isolates appeared after the cessation of treatment and maintained throughout the experiments. On the contrary, our results of study 2 demonstrated that administration of enrofloxacin generated a rapid increase of fluoroquinolone resistance in C. jejuni showing the mutation of Asp 90-Asn in the gyrA gene. Present results indicate that the enrofloxacin treatment for broilers at the inherent dosage is able to select fluoroquinolone resistance in C. jejuni. We conclude that whatever enrofloxacin dosage is used, an emergence of fluoroquinolone resistant of C. jejuni occurs. PMID- 16364023 TI - Development of performance indicators for the bovine clinical salmonellosis surveillance network in France. AB - Performance indicators are variables used to calculate on a continuous basis the operational level of a surveillance network's priority activities. A 10-step process was developed to enable network coordinators to identify specific performance indicators to help them monitor their networks. The methodology was based on a listing of surveillance activities, the definition and choice of network's global priority objectives, the construction of performance indicators and monitoring tables and the implementation of the system within the network. This process was implemented for the bovine clinical digestive salmonellosis surveillance network in France (RESSAB). The process produced a list of 26 activities synthesized into 15 global objectives, from which 12 were retained as priority objectives. This made it possible to develop 12 performance indicators. Indicators retrospectively calculated for the year 2003 indicated that RESSAB was operating according to the objectives set by the network's stakeholders and its financial supporter. The methodology was implemented successfully and was not very time consuming (12 person-days) or expensive. The decision makers and actors in the network quickly appropriated the system. The methodology can thus be considered validated through this example. Nevertheless, the risks inherent in the use of performance indicators must be addressed by ensuring the relevance of the selected indicators through external assessment and by prioritizing an internal and participatory approach to avoid a misuse of the performance indicators. In addition, considering that performance indicators address only the operation of the network, relevance of surveillance procedure should be addressed by external evaluation. PMID- 16364024 TI - Occurrence of salmonella in the ileum, ileocolic lymph nodes, tonsils, mandibular lymph nodes and carcasses of pigs slaughtered for consumption. AB - This study evaluates the occurrence of Salmonella in pork carcasses and in some risk tissues (ileum, ileocolic and mandibular lymph nodes and tonsils), that can be involved in Salmonella contamination during slaughter. Salmonella was identified in 27 (26.7%) pigs and in 13 (12.9%) carcasses. From these positive carcasses, 69.2% presented the same serotype as that identified in the corresponding pig, which emphasize the pigs importance as a source of Salmonella during the slaughter, suggesting that measures should be taken at the level of pig production in order to reduce the slaughtering of Salmonella-positive animals. The highest value of Salmonella occurrence was reached in the ileocolic lymph nodes (18.8%) and in the ileum (13.9%), representing Salmonella potential faecal source during pork processing at the abattoir. In these samples, a high level of Salmonella was observed in the ileocolic lymph nodes in comparison with the ileum. The mandibular lymph nodes (12.9%) also presented a higher occurrence in comparison with the tonsils (9.9%). These results indicate that the lymph nodes analysis could be more sensitive in the detection of Salmonella than the closer drainage tissue. Otherwise, the presence of Salmonella in the lymph nodes indicates lymphatic spread of the organism, which reflects an increased risk of pork contamination. These results also indicate that, in order to achieve a better control of Salmonella contamination during the slaughter process, it is important to consider the improvement of the evisceration practices and the tonsils as well the extraction of mandibular lymph nodes after slaughter. PMID- 16364025 TI - Prevalence of Cryptosporidium spp. in asymptomatic sheep in family flocks from Mexico State. AB - To determine Cryptosporidium spp. prevalence in faeces from asymptomatic sheep, 37 family flocks in Mexico State were sampled. Data were collected by the interview method and Ziehl-Neelsen stain of faecal samples for Cryptosporidium identification. The independent group proportion hypothesis test was used for differences in prevalences between sheep farms. A 34.3% general prevalence was found, and prevalence in herds with >100 animals was significantly higher (40.6%). Correlation was found between flock size and Cryptosporidium spp. presence (odds ratio, OR, 2.57; confidence interval, IC(95%), 1.84-3.60). PMID- 16364026 TI - Hair growth inhibition by psychoemotional stress: a mouse model for neural mechanisms in hair growth control. AB - Stress has long been discussed controversially as a cause of hair loss. However, solid proof of stress-induced hair growth inhibition had long been missing. If psychoemotional stress can affect hair growth, this must be mediated via definable neurorendocrine and/or neuroimmunological signaling pathways. Revisiting and up-dating relevant background data on neural mechanisms of hair growth control, we sketch essentials of hair follicle (HF) neurobiology and discuss the modulation of murine hair growth by neuropeptides, neurotransmitters, neurotrophins, and mast cells. Exploiting an established mouse model for stress, we summarize recent evidence that sonic stress triggers a cascade of molecular events including plasticity of the peptidergic peri- and interfollicular innervation and neuroimmune crosstalk. Substance P (SP) and NGF (nerve growth factor) are recruited as key mediators of stress-induced hair growth-inhibitory effects. These effects include perifollicular neurogenic inflammation, HF keratinocyte apoptosis, inhibition of proliferation within the HF epithelium, and premature HF regression (catagen induction). Intriguingly, most of these effects can be abrogated by treatment of stressed mice with SP-receptor neurokinin-1 receptor (NK-1) antagonists or NGF-neutralizing antibodies - as well as, surprisingly, by topical minoxidil. Thus there is now solid in vivo-evidence for the existence of a defined brain- HF axis. This axis can be utilized by psychoemotional and other stressors to prematurely terminate hair growth. Stress induced hair growth inhibition can therefore serve as a highly instructive model for exploring the brain-skin connection and provides a unique experimental model for dissecting general principles of skin neuroendocrinology and neuroimmunology well beyond the HF. PMID- 16364027 TI - Defining the caspase-containing apoptotic machinery contributing to cornification in human epidermal equivalents. AB - Whether terminal differentiation/stratum corneum formation of keratinocytes (KCs) represents a form of programmed cell death, utilizing mediators of classical apoptosis, is unclear. Apoptosis, an evolutionarily conserved death process, is comprised of extrinsic and intrinsic pathways, which converge using caspase 3. To define upstream and downstream caspases involved in terminal differentiation, we utilized human epidermal equivalents (EEs). Using submerged cultures comprised of human KCs, EEs were sequentially analyzed before and after being raised to an air/liquid (A/L) interface at 3-24 h intervals. At each time point, EEs were analyzed morphologically and for specific enzyme activity to distinguish different initiator (caspases 1, 2, 8, 9) and effector caspases (3, 6, 7). Terminal differentiation began at 6-8 h, as defined by stratum corneum with loricirin expression and completed at 18-24 h producing an epidermis resembling normal skin. Enzyme activity for caspases 1, 2, 3, 6, 7, 8, and 9 (but not 4, 5) was enhanced (>two-fold nmol/mg/h) at 3-6 h compared with submerged cultures. Processing of caspase 14 occurred at 18 h, and cleaved caspase 14 was increased at 24 h. Activated caspase 3-positive and terminal deoxynucleotidyl transferase mediated nick end labeling-positive KCs were identified in EEs at 3-6 h corresponding to initiation sites of terminal differentiation. Addition of caspase inhibitors reduced levels of involucrin and loricrin in EEs raised to an A/L interface. We conclude caspases function as important death effectors strategically positioned at intersection of intrinsic and extrinsic pathways in KCs undergoing stratum corneum formation. PMID- 16364028 TI - Apoptosis induced by ultraviolet B in HPV-immortalized human keratinocytes requires caspase-9 and is death receptor independent. AB - Ultraviolet B (UVB) induces both apoptosis and skin cancer. We found that human keratinocytes (KC) immortalized by Human Papillomavirus (HPV)16 E6/E7 were sensitized to UVB-induced apoptosis, possibly representing a transient regression prone precancerous stage equivalent to actinic keratosis. To further examine which caspases are apical and essential, we utilized retroviral constructs expressing dominant-negative caspase-9 (caspase-9-DN) or Fas-associated protein with death domain (FADD)-DN as well as caspase inhibitor peptides. Caspase-9-DN and zLEHD-fmk both suppressed caspase-9, -3, and -8 activity after UVB exposure, as well as proteolytic processing of procaspase-3 into its active form, DNA fragmentation factor 45 cleavage, and internucleosomal DNA fragmentation. By contrast, stable expression of FADD-DN in HPV-immortalized KC did not inhibit UVB induced activation of caspases-9, -3, and -8 nor downstream apoptotic events, although inhibition of caspase-8 with zIETD-fmk attenuated apoptosis. This study indicates that caspase-9 activation is upstream of caspases-3 and -8 and that UVB induced apoptosis in HPV-immortalized human KC is death receptor (DR) independent and requires both caspase-9 upstream and caspase-8 downstream for maximal apoptosis. These studies further indicate that cell type as well as transformation state determine the sensitivity and mode of cell death (DR vs. mitochondrial apoptotic pathways) in response to UVB and explain the high regression rates of premalignant lesions. PMID- 16364029 TI - Expression of matrix metalloproteinase (MMP)-2, MMP-9, MMP-13, and MT1-MMP in skin tumors of human papillomavirus type 8 transgenic mice. AB - Human papillomaviruses (HPV) are small DNA viruses that induce a wide variety of hyperproliferative lesions in cutaneous and mucosal epithelia. It is proposed that HPV is involved in non-melanoma skin cancer development. We have previously shown that HPV8 transgenic mice spontaneously develop papillomatous skin tumors. Histology revealed epidermal hyperplasia, acanthosis and hypergranulosis and in some cases squamous cell carcinomas (SCC). Zymographic and immunoblot analysis of normal skin extracts identified increased amounts of matrix metalloproteinase (MMP)-9, MMP-13 and MT1-MMP in HPV8-positive mice compared with HPV8-negative animals. In situ gelatin zymography of tumor specimens displayed a strong proteolytic activity in papillomas, and SCC putatively attributed to the increased amounts of activated MMP-9 found in tissue extracts. In addition, immunoblot analysis revealed increased amounts of active MMP-13 and MT1-MMP in tumor extracts as compared with control extracts. Immunohistochemical stainings of SCC specimens depicted MMP-13 to be specifically expressed in stromal fibroblasts neighboring the tumor islands, whereas MT1-MMP was detected both in tumor cells and in stromal cells. Taken together, these results implicate a role for MMPs in the development of HPV8-induced cutaneous tumors. PMID- 16364030 TI - Effect of pimecrolimus vs. corticosteroids on murine bone marrow-derived dendritic cell differentiation, maturation and function. AB - Pimecrolimus (SDZ ASM981) is a non-steroid member of calcineurin inhibitors recently developed for the treatment of inflammatory skin diseases. In this study, we compared the effect of pimecrolimus and corticosteroids on the differentiation, maturation and function of murine bone marrow-derived dendritic cells (BM-DC). We added pimecrolimus at concentrations of 5-500 ng/ml or 0.5 ng/ml mometasone furoate at different timepoints to the BM-DC culture and checked (i) the number of matured cells, (ii) the expression of activation markers, (iii) the release of cytokines and (iv) the stimulatory capacity of the resulting BM-DC in vivo. Even at the highest concentration, pimecrolimus treatment resulted in only modest effects. In the pimecrolimus-treated culture, we observed a decrease in the numbers of matured cells but no significant effects on the expression of activation markers. The release of some inflammatory cytokines was reduced, but the stimulatory capacity in vivo was not affected. In contrast, mometasone furoate has pronounced effects on BM-DC at a concentration ten to 1000 times lower than those used with pimecrolimus. Furthermore, topical treatment of mice with clobetasole cream 0.05% resulted in almost complete depletion of splenic DC and a severe hyposplenia, while high-dose oral pimecrolimus treatment did not show any effects on the spleen or on splenic DC. These results support that pimecrolimus, unlike corticosteroids, has little effects on dendritic cells. To the best of our knowledge, this is the first study of this type with use of BM DC. PMID- 16364031 TI - Transduced monocyte/macrophages targeted to murine skin by UV light. AB - We have selectively targeted monocyte/macrophages overexpressing an immunomodulatory molecule, latency-associated peptide (LAP), a naturally occurring antagonist for transforming growth factor-beta1, to murine skin utilizing UV light to produce a cutaneous influx of transduced monocyte/macrophages. Bone marrow (BM) cells from BALB/c mice were transduced in vitro with a retroviral construct containing green fluorescent protein (GFP) for detection and human LAP (hLAP) as a test molecule. The transduced BM cells were then cultured in vitro with granulocyte-macrophage colony-stimulating factor (GM CSF) to produce differentiation to monocyte/macrophages. More than 80% of transduced BM cells were CD11b-positive and MOMA-positive by fluorescence activated cell-sorter analysis and secreted LAP by ELISA after 10 days of culture in granulocyte-macrophage colony-stimulating factor (GM-CSF). Transduced monocyte/macrophages containing either GFP or hLAP-GFP were then injected intravenously into BALB/c mice. One-half of recipients in each group were exposed to UVB (72 mJ) to induce monocyte/macrophage infiltration into skin. Recipients were sacrificed 60 h after UV irradiation. We found transduced cutaneous macrophages expressing GFP by examining with fluorescence microscopy frozen skin sections of recipient mice immunostained with antibodies to GFP and to macrophage marker F4/80. We identified hLAP sequences by polymerase chain reaction (PCR) of total DNA in recipient blood and UV-irradiated skin but not in unirradiated skin. LAP sequences were also detected at much lower levels in other organs (lung, spleen, and liver) by PCR. Therefore, we have shown that genetically altered monocytic cells can be injected intravenously and targeted to mouse skin by UV exposure. This macrophage-based gene-transfer method may be a potentially useful immunotherapeutic approach for delivering monocyte/macrophage-derived products to skin. PMID- 16364032 TI - Neurogenic components of trypsin- and thrombin-induced inflammation in rat skin, in vivo. AB - Activation of protease-activated receptors (PAR) can induce vasodilation (VD) and increase of vascular permeability either directly by stimulating endothelial cells or indirectly via activation of nociceptors and subsequent release of neuropeptides (neurogenic inflammation). We aimed to estimate the relative contribution of the two pathways for stimulation with endogenous activators of PAR-2 (trypsin) and of PAR-1, 3 and 4 (thrombin) using in vivo dermal microdialysis in rats. Protein extravasation (PE) was assessed by increase of protein concentration in the dialysate, and VD was quantified by laser Doppler scanning. Both trypsin (10(-8)-10(-4) M) and thrombin (10(-6), 10(-5.5) and 10( 5) M) provoked PE and local VD in a dose-dependent manner. Trypsin (10(-4) M) induced PE was inhibited by 87.2 +/- 21% due to the substance P (SP) NK1 receptor antagonist SR140333. VD was blocked by 58.15 +/- 10.1% in response to the calcitonin gene-related peptide (CGRP) receptor antagonist CGRP(8-37). By contrast, CGRP(8-37) did not affect thrombin-induced VD, while blockade of SP receptors prevented the PE elicited only by low doses of thrombin (10(-6) M), being ineffective at higher thrombin concentrations. In conclusion, intradermal trypsin elicits a neurogenic inflammation in rat, probably mediated via PAR-2 activation on nociceptors and subsequent SP and CGRP release. Thrombin-induced PE and VD are mediated mainly by a non-neurogenic mechanism. PMID- 16364033 TI - Phytocomponents of triterpenoids, oleanolic acid and ursolic acid, regulated differently the processing of epidermal keratinocytes via PPAR-alpha pathway. AB - Naturally occurring triterpenoids such as oleanolic acid (OA) and ursolic acid (UA) are known to have anti-inflammatory and anticarcinogenic activities in some types of cells. Although it has been reported that UA increases the amount of ceramide in keratinocytes, there is little study on the mechanism of triterpenoids involved in the differentiation of keratinocytes as well as their effects on epidermal permeability barrier. A study was therefore conducted to determine whether OA and UA could stimulate the differentiation of epidermal keratinocytes through peroxisome proliferator-activated receptor (PPAR)-alpha activation. This work was then extended to investigate the rate of formation of cornified envelope as a marker in the terminal differentiation of keratinocytes and the amount of transglutaminase in human keratinocytes treated with OA and UA. It was shown that OA induced the differentiation of keratinocytes, whereas UA had little effect. In addition, reporter gene assay using PPAR response element activity demonstrated that OA might be related to the increase of PPAR-alpha activity in CV-1 cells. Moreover, it enhanced the recovery of epidermal permeability barrier function as well as increased ceramides in epidermis after topical application. We therefore propose that the effect of OA on the stimulation of differentiation in epidermal keratinocytes seems to be highly related to activation of PPAR-alpha. PMID- 16364034 TI - Dietary constituents are able to play a beneficial role in canine epidermal barrier function. AB - Epidermal barrier function is a critical attribute of mammalian skin. The barrier is responsible for preventing skin-associated pathologies through controlling egress of water and preventing ingress of environmental agents. Maintaining the quality and integrity of the epidermal barrier is therefore of considerable importance. Structurally, the barrier is composed of two main parts, the corneocytes and the intercellular lamellar lipid. The epidermal lamellar lipid comprises mainly ceramides, sterols and fatty acids. Twenty-seven nutritional components were screened for their ability to upregulate epidermal lipid synthesis. Seven of the 27 nutritional components (pantothenate, choline, nicotinamide, histidine, proline, pyridoxine and inositol) were subsequently retested using an in vitro transepidermal diffusion experimental model, providing a functional assessment of barrier properties. Ultimately, the best performing five nutrients were fed to dogs at supplemented concentrations in a 12-week feeding study. Barrier function was measured using transepidermal water loss (TEWL). It was found that a combination of pantothenate, choline, nicotinamide, histidine and inositol, when fed at supplemented concentrations, was able to significantly reduce TEWL in dogs after 9 weeks. PMID- 16364035 TI - Nonsurgical factors affecting symptomatic outcomes of antireflux surgery. AB - A small number of patients will have persistent or new symptoms after antireflux surgery for gastroesophageal reflux disease (GERD). Most of these symptoms are due to recurrent reflux or some complication or side-effect of the operation. However, a few of these patients will be symptomatic without objective findings to explain these symptoms. The purpose of this review is to highlight potential non-surgical factors that may proceed to a poor symptomatic outcome after antireflux surgery. These factors include underlying esophageal pathophysiology, issues related to chronic pain and pain perception, personality and psychoemotional disorders, functional esophageal and/or bowel disorders, and the nocebo phenomenon. Awareness of these other causes can lead to more appropriate treatments. PMID- 16364036 TI - Serum concentrations of high-mobility group box chromosomal protein 1 before and after exposure to the surgical stress of thoracic esophagectomy: a predictor of clinical course after surgery? AB - High-mobility group box chromosomal protein 1 (HMGB-1) has recently been shown as an important late mediator of endotoxin shock, intra-abdominal sepsis, and acute lung injury. However, its role in the systemic inflammatory response syndrome after major surgical stress, which may lead to multiple organ dysfunction syndrome, has not been thoroughly investigated. We hypothesized that serum HMGB-1 participates in the pathogenesis of postoperative organ system dysfunction after exposure to major surgical stress. A prospective clinical study was performed to consecutive patients (n = 24) with carcinoma of the thoracic esophagus who underwent transthoracic esophagectomy with three field lymph node resection between 1998 and 2003 at Keio University Hospital, Japan. Serum HMGB-1 concentrations were measured by enzyme-linked immunosorbent assay. Preoperative serum HMGB-1 levels correlated with postoperative duration of SIRS, mechanical ventilation, and intensive care unit stay. Three of the 24 patients had serious postoperative complications: sepsis in two, and acute lung injury in one. Serum HMGB-1 levels in patients without complications increased within the first 24 h postoperatively, remained high during postoperative days 2-3, and then decreased gradually by postoperative day 7. In patients with serious complications, serum HMGB-1 was significantly higher than that found in patients without postoperative complications at every time point except postoperative day 2. Preoperative serum HMGB-1 concentration seems to be an important predictor of the postoperative clinical course. Transthoracic esophagectomy induces an increase in HMGB-1 in serum even in patients without complications. Postoperative serum HMGB-1 concentrations were higher in patients who developed complications, and may be a predictive marker for complications in this setting. PMID- 16364037 TI - Comparative genomic hybridization of esophageal adenocarcinoma and squamous cell carcinoma cell lines. AB - We compared whole genomic changes in cell lines generated from esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). To do so, we investigated chromosomal DNA copy number changes in four EAC cell lines and three ESCC cell lines using comparative genomic hybridization (CGH). Frequent gains of chromosome 5p, 8q, and 20q occurred in both ESCC and EAC cell lines, but gains of 3q, 5q, and 9q were mainly seen in ESCC cell lines; gain of chromosome 10q25-qtel was mainly seen in EAC cell lines. It was noticeable that 18q12 loss existed in 2 EAC and 1 ESCC cell lines in our study. The chromosomal abnormalities common to all of the cell lines may help to identify candidate genes related to both EAC and ESCC. The chromosome aberrations mainly seen in either EAC or ESCC cell lines are in keeping with their known different etiology and may lead to the identification of genes important for disease specific pathogenesis. PMID- 16364038 TI - Radiation therapy combined with cis-diammine-glycolatoplatinum (nedaplatin) and 5 fluorouracil for Japanese stage II-IV esophageal cancer compared with cisplatin plus 5-fluorouracil regimen: a retrospective study. AB - To evaluate the treatment outcome of radiotherapy combined with cis-diammine glycolatoplatinum (nedaplatin) plus 5-fluorouracil (5-FU) for esophageal cancer. From January 2000 to December 2004, a total of 12 esophageal cancer patients with locally advanced and metastatic esophageal cancer (stages II-IVB) were treated with radiation therapy (50.4 Gy) combined with nedaplatin (80 mg/m(2), bolus infusion) and 5-FU (800 mg/m(2)/24 h, continuous infusion for 4 days) (NDP group). We compared the data with those of patients during the same period receiving a different chemotherapy regimen consisting of cisplatin (75 mg/m(2), bolus infusion) and 5-FU (1000 mg/m(2)/24 h, continuous infusion for 4 days) (n = 29, CDDP group) combined with the same radiation therapy. The median survival period was 11.5 months in the NDP group and 13.1 months in the CDDP group. The overall survival rates at 1-, 2-, and 3-years were 40%, 13%, and 13% in the NDP group and 56%, 42%, and 8% in the CDDP group (P = 0.2472), respectively. Grade III and IV leukocytopenia was observed in six (50%) and none of the patients in the NDP group and 14 (48%) and seven (24%) in the CDDP group, respectively. Grade III thrombocytopenia was observed in three (25%) in the NDP group and four (14%) in the CDDP group. Radiation combined with nedaplatin and 5-FU is a safe and effective method for treating esophageal cancer. We recommend that NDP should be used rather than dose-reduction of CDDP combined with 5-FU in patients with impaired renal function as indicated by low creatinine clearance value (40-60 mL/min). PMID- 16364039 TI - Genes associated with telomerase activity levels in esophageal carcinoma cell lines. AB - Telomerase activity levels have been shown to correlate with tumor progression in several malignancies. However, the genetic regulation of telomerase activity levels is not fully understood. The aim of the present study has been to identify a gene expression profile, predicting correlation with the telomerase-activity test. Ten human esophageal carcinoma cell lines were investigated using the telomerase activity assay (TRAPeze) Telomerase Detection Kit), followed by further characterization using the GeneChip Human Genome U133A 2.0 Array (Affymetrics Inc., USA), including 14 500 human genes. Telomerase activity levels were detected in all cell lines with a broad range of activity levels. Using a high correlation coefficient, r > 0.90, the following genes were found to be positively correlated with telomerase activity levels: N-myristoyltransferase 2; ribosomal protein L3; retinoblastoma-like 2 (pRb2/p130); and cyclin G2. Only one gene was negatively correlated with telomerase activity levels, zinc finger protein 207. In conclusion, the present microarray data provide primary validation data indicating possible candidates for prognostic and prediction factors in esophageal cancer in relation to telomerase activity. PMID- 16364040 TI - Thoracic esophageal perforation: one surgeon's experience. AB - Recognition of the importance of early diagnosis and aggressive, definitive surgical intervention has brought about a dramatic decline in mortality related to distal esophageal perforation. In the following retrospective analysis, we have examined all cases of thoracic esophageal perforations diagnosed, consulted, and/or treated by one author (PDK) at the Inova Fairfax Hospital from June 1, 1988 through March 17, 2005. These cases consisted of 48 patients (34 male) with a mean age of 59.4 years (range, 20-92). Among 25 patients with early diagnosis (< or = 24 h), hospital survival was 92%, increasing to 96% when early diagnosis was combined with surgical treatment. Among the 23 patients with late diagnosis (> 24 h), hospital survival was 82.6%, increasing to 92.3% when treated with surgery. We recommend aggressive, definitive surgery for thoracic esophageal perforations, regardless of time of diagnosis. In the absence of phlegmon or implacable obstruction, primary repair offers excellent results with the shortest length of stay. Resection and reconstruction are the best choices in circumstances where significant phlegmon or distal obstruction render primary repair hazardous or inapplicable. Diversion, preferably with proximal and distal esophageal exclusion, may be necessary for patients too ill to undergo more formidable surgery. Conservative, medical therapy may be appropriate in patients with 'microperforations' with no continuing leak. Finally, comfort measures alone may be appropriate where circumstances merit no effort at resuscitation. PMID- 16364041 TI - Lower esophageal sphincter analysis using computerized manometry in patients with chagasic megaesophagus. AB - Due to the introduction of computer technology into manometry laboratories, three dimensional manometric images of the lower esophageal sphincter can be constructed based on radially oriented pressures, a method termed 'computerized axial manometry.' Calculation of the sphincter pressure vector volume using this method is superior to standard manometric techniques in assessing lower esophageal sphincter function in patients with gastroesophageal reflux disease and idiopathic achalasia. Despite similarities between idiopathic achalasia and chagasic esophagopathy found using clinical, radiological, and manometric studies, controversy around lower esophageal sphincter pressure persists. The goal of this study was to analyze esophageal motor disorders in Chagas' megaesophagus using computerized axial manometry. Twenty patients with chagasic megaesophagus (5 men, 15 women, and average age 50.1 years, range 17-64) were prospectively studied. For three-dimensional imaging construction of the lower esophageal sphincter, a low-complacency perfusion system and an eight-channel manometry probe with four radial channels placed in the same level were used. For probe traction, the continuous pull-through technique was used. Results showed that the lower esophageal sphincter of patients with chagasic megaesophagus have significantly elevated pressure, length, asymmetry, and vector volumes compared to those of normal volunteers (P < 0.05). Aperistalsis of the esophageal body waves was observed in all patients and contraction amplitude was lower than that in normal patients. We conclude that patients with chagasic megaesophagus have hypertonic lower esophageal sphincter and aperistalsis of the esophageal body. PMID- 16364042 TI - Tracheo-esophageal fistula: combined surgical and endoscopic approach. AB - We present a case of benign acquired tracheo-esophageal fistula caused by cuffed tracheal tube. The patient was septic and weaning from the mechanical ventilation was unlikely, so immediate repair of the tracheal and esophageal lesions was imperative. A silicone endoprosthesis was inserted in the tracheal lumen and the esophageal lesion was repaired with direct suture. After weaning from mechanical ventilation and recovery, the patient was discharged with a silicone tracheal stent. Three months later the stent was removed without complications or need for further treatment. This combined approach as never been published before, and can be a very useful alternative for the treatment of this rare but taxing complication. PMID- 16364043 TI - Diaphragmatic acute massive herniation after laparoscopic gastroplasty for esophagectomy. AB - Minimally invasive techniques are increasingly being used for oesophagectomy. Diaphragmatic hernia is a rare complication of gastroplasty in open surgery. One of the advantages of the laparoscopic technique, the lack of peritoneal adhesions, may lead to an increased rate of this complication. We report two cases of diaphragmatic acute massive herniation after laparoscopic gastroplasty for esophagectomy out of a series of 44 laparoscopic gastroplasties performed over 33 months. We discuss some technical aspects related to its occurrence. Prevention should include a limited crural division and fixation of the gastric tube to the diaphragmatic crura at primary surgery. PMID- 16364044 TI - Postpneumonectomy esophageal fistula: a combined radiological-endoscopic technique. AB - We report a case of a 70-year-old man who presented with a long-standing esophagocutaneous fistula following a pneumonectomy for aspergilloma. Major surgical procedures, including a pectoralis major flap reconstruction, a pedicled omental transposition and a radial forearm flap transposition, failed to obliterate the fistula. Seven years after initial surgery the esophagocutaneous fistula was successfully treated by means of a minimally invasive joint endoscopic and radiological technique. A radiographic catheter was passed through the fistula. The catheter and the guide wire were manipulated into the esophageal defect and into the upper esophagus. Under endoscopic vision, the catheter was then advanced over the guide wire and out of the patient's mouth. A T-tube was sutured to the catheter outside the mouth, pulled down through the esophagus, and into the esophageal defect and out through the chest wall, leaving the T-part of the tube within the esophagus. The patient made a good recovery and was discharged 7 days later. He was able to resume oral intake 3 weeks after the procedure. PMID- 16364045 TI - True carcinosarcoma of the esophagus. AB - Most esophageal carcinosarcomas are diagnosed as so-called carcinosarcoma, in which individual elements may be derived from a single common ancestor cell, and there have been a few reports describing true carcinosarcoma originating from two individual stem cells. We describe a case of esophageal carcinosarcoma exhibiting neoplastic osteoid formation. Immunoreactivity for vimentin and p53 was limited to only the sarcomatous component and was absent in the carcinomatous component. Furthermore, a point mutation in exon 7 of the p53 gene was observed only in the sarcomatous component. Both sarcoma and carcinoma cells distinctively metastasized to different lymph nodes. These observations led us to diagnose the esophageal tumor as a true carcinosarcoma. PMID- 16364046 TI - A missing denture's misadventure! AB - We report a late onset, benign, tracheoesophageal fistula in a 51-year-old man, due to an accidentally swallowed denture. In view of the extensive peri esophageal sepsis and fibrosis, he was managed by a subtotal esophagectomy and a cervical esophagogastric anastomosis. The tracheal defect was closed with the help of an intercostal muscle flap. This report also highlights the difficulty in identifying swallowed prosthetic dental material radiologically, when no metallic component is present. This fact was also responsible for the delay in diagnosis, eventually leading to the rare complication of a tracheoesophageal fistula. PMID- 16364048 TI - Pathophysiology and treatment of acute pancreatitis: new therapeutic targets--a ray of hope? AB - Acute pancreatitis is a life-threatening disease with putatively high mortality rates, particularly in the setting of systemic inflammatory response and multiple organ failure when superinfection of necrosis occurs. Although the APACHE II and Ranson score are widely accepted as clinical scores to predict the prognosis, current medical treatment is still based upon state of the art intensive care treatment largely unrelated to the pathogenesis of the disease. The mechanisms by which premature enzyme activation and autodigestion of the pancreatic gland is triggered and maintained are still ill-defined. It is well known that activation of chemokines, cytokines and pancreatic enzymes characterize the cause of the disease, but disease-phase specific treatment attempts have thus far not resulted in successful molecular based medical treatments. The current summary describes the novel understanding in the pathophysiology of acute pancreatitis with special emphasis on specific disease phases. It outlines promising and novel experimental and medical therapeutic approaches which might become clinical targets and successful strategies to significantly reduce pancreatitis-associated mortality rates. PMID- 16364047 TI - Pharmacological targets in gastro-oesophageal reflux disease. AB - Although proton pump inhibitors have become the mainstay of treatment in gastro oesophageal reflux disease (GORD), there are still unmet needs in the management of this very common disorder. For example, all current proton pump inhibitors have a relatively slow onset of action and their activity is limited mainly to the post-prandial period with far less effective inhibition of nocturnal acid secretion. In order to achieve more potent, rapid and sustained acid inhibition several compounds are currently under development, such as new proton pump inhibitors with a prolonged plasma half-life, potassium competitive ATPase blockers (PCABs), histamine H3 agonists, and gastrin antagonists. Acid suppression does not, however, cure the disease and relapses are frequently observed after discontinuation of proton pump inhibitor therapy. Among the different abnormalities involved in the pathophysiology of this multifactorial disease, transient lower oesophageal sphincter relaxations represent the major mechanism responsible for episodes of reflux. Baclofen, the prototype GABA(B) receptor agonist, is one of the most potent inhibitors of transient lower oesophageal sphincter relaxations identified. To date the transfer of these relaxation-controlling pharmacological agents into clinical practice has however been hampered by the occurrence of unacceptable side effects. Beside "anti relaxation therapy", the potential of novel prokinetics such as motilin agonists has been explored, especially since the motilin receptor has been cloned. Thus far the broad therapeutic value of prokinetics in GORD does, however, seem very limited in terms of efficacy with respect to oesophageal motility and acid exposure. Lastly, further research is necessary to better understand the complex mechanisms involved in oesophageal sensitivity and mucosal defence. PMID- 16364049 TI - Pharmacokinetics of esomeprazole following varying intravenous administration rates. AB - There are situations where the use of an oral proton pump inhibitors is not possible. In such situations an intravenous route is the preferred alternative. An intravenous formulation of esomeprazole has recently been developed. This study was designed to evaluate the pharmacokinetics and tolerability of single dose intravenous esomeprazole using different rates of administration. The study was an open randomised, cross-over design in healthy male and female (n = 24). Esomeprazole 40 mg intravenously was administrated as an infusion over 10, 15, 20 or 30 min., or esomeprazole 20 mg intravenously as an injection over 3 min. There was a wash-out period of at least 6 days between dose regimens. It was demonstrated that increasing the rate of intravenous infusion of esomeprazole 40 mg resulted in higher Cmax values (geometric means; 5.2-7.6 micromol/l), but the AUC values remained relatively constant (7.1-7.2 micromor/l). As expected esomeprazole 20 mg administered as a 3 min. intravenous injection had lower Cmax (3.6 micromol/l) and AUC (2.9 micromol.r/l) values than any of the infusions of esomeprazole 40 mg. Intravenous esomeprazole was well tolerated in this study. In conclusion, any variation in the infusion rate of esomeprazole 40 mg intravenously has little effect on the pharmacokinetics of esomeprazole in healthy volunteers, which provides flexibility in the choice of dosing regimens. PMID- 16364050 TI - Expression of inducible nitric oxide synthase in trigeminal ganglion cells during culture. AB - Nitric oxide (NO) is an important signalling molecule that has been suggested to be a key molecule for induction and maintenance of migraine attacks based on clinical studies, animal experimental studies and the expression of nitric oxide synthase (NOS) immunoreactivity within the trigeminovascular system. Sensitisation of the trigeminal system including the trigeminal ganglia neurones is believed to be involved in the pathway leading to migraine pain. In the present study, the NOS expression in rat primary trigeminal ganglia neurones was examined at different time points using immunocytochemistry, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting. In trigeminal ganglia cells not subjected to culture, endothelial (e) and neuronal (n) but not inducible (i) NOS mRNA and protein were detected. Culture of rat neurones resulted in a rapid axonal outgrowth of NOS positive fibres. At 12, 24 and 48 hr of culture, NOS immunoreactivity was detected in medium-sized trigeminal ganglia cells. Western blotting and RT-PCR revealed an up-regulation of inducible iNOS expression during culture. However, after culture only low levels of eNOS protein was found while no eNOS and nNOS mRNA and protein could be detected. The data suggest that iNOS expression may be a molecular mechanism mediating the adaptive response of trigeminal ganglia cells to the serum free stressful stimulus the culture environment provides. It may act as a cellular signalling molecule that is expressed after cell activation. PMID- 16364051 TI - Effects of concurrent exposure to tributyltin and 1,1-dichloro-2,2 bis (p chlorophenyl) ethylene (p,p'-DDE) on immature male Wistar rats. AB - Tributyltin and 1, 1-dichloro-2, 2 bis (p-chlorophenyl) ethylene (p,p'-DDE) have been ubiquitously distributed over the world. In Japan, p,p'-DDE and tributyltin are ingested through marine products, in which these substances are accumulated through bio-concentration and the food chain. However, the consequence of potential combined hazards of these substances remains unknown. Therefore, the effects of concurrent exposure to 125 ppm p,p'-DDE and 25 ppm tributyltin were investigated in immature male Wistar rats by oral administration during puberty. In this study, tributyltin promoted the growth of pubertal male rats, while p,p' DDE itself did not affect the growth but inhibited the growth enhancement by tributyltin. Furthermore, tributyltin reduced thymus weight but p,p'-DDE also prevented this weight reduction. Neither development of male sexual accessory organs nor sexual maturation was affected even by concurrent exposure to p,p'-DDE and tributyltin. No significant changes of serum testosterone, luteinizing hormone, follicle-stimulating hormone concentrations, and epididymal sperm numbers were observed with the administration of p,p'-DDE and/or tributyltin. These results indicate that sexual maturation, male reproductive organ development and sperm production is scarcely affected in immature male Wistar rats even by concurrent exposure to p,p'-DDE and tributyltin at a daily dose of ca. 2 mg/kg tributyltin and 10 mg/kg p,p'-DDE. Moreover, the simultaneous administration of p,p'-DDE with tributyltin counterbalanced the effects that were attributed to tributyltin alone. PMID- 16364052 TI - Effects of acute administration of and tachyphylaxis to alpha,beta-methylene ATP in the guinea-pig small intestine. AB - The aim of the present study was to assess the acute motility effects and desensitizing activity of the stable ATP analogue and P(2X) purinoceptor agonist alpha,beta-methylene ATP (alpha,beta-meATP) and the effect of alpha,beta-meATP desensitization on nerve-mediated cholinergic responses in the guinea-pig ileum in vitro. It was confirmed that alpha,beta-meATP (1-30 microM) causes neurally mediated, cholinergic (tetrodotoxin- and atropine-sensitive) longitudinal contractions. These responses were not influenced by the ganglionic blocking drug hexamethonium (50 microM), or a combination of the adrenergic neurone blocking drug guanethidine (3 microM), the opioid receptor antagonist naloxone (0.5 microM) and the nitric oxide synthase inhibitor N(G)-nitro-L-arginine (L-NOARG; 100 microM), but were strongly reduced or abolished by the P2 purinoceptor antagonist PPADS (30 microM) or by tachyphylaxis evoked by 10 microM alpha,beta meATP. The contractile effect of alpha,beta-meATP (3 microM) was moderately inhibited by 10 microM and strongly suppressed by 30 microM of NF 279, an antagonist predominantly affecting P2X1 purinoceptors, but left uninfluenced by the P2X(5,7) receptor antagonist Brilliant blue G. No relaxant effect of alpha,beta-meATP was detected in the concentration range of 1-30 microM. Tachyphylaxis to alpha,beta-meATP (1-10 microM) caused a moderate inhibition of the cholinergic (atropine-sensitive) contractile response of the ileum to electrical field stimulation (5 Hz for 5 sec.). This reduction was unaltered in the presence of guanethidine, naloxone and L-NOARG. Responses to nicotine (1 or 2 microM) were not reduced by alpha,beta-meATP tachyphylaxis. It is suggested that alpha,beta-meATP-sensitive P(2X) purinoceptors are involved in the prejunctional modulation of cholinergic neurotransmission between the myenteric plexus and longitudinal smooth muscle in the guinea-pig small intestine. PMID- 16364053 TI - Acute renal effects of intravenous bisphosphonates in the rat. AB - Bisphosphonates are potent osteoclast inhibitors that have been associated with renal toxicity following rapid intravenous administration of high doses, which was hypothesised to be due to precipitation of bisphosphonate aggregates or complexes in the kidney. Five studies were conducted in rats investigating the characteristics of bisphosphonate-related acute renal effects. These studies included single intravenous injections of the nitrogen-containing bisphosphonates (1) ibandronate (1-20 mg/kg), or (2) zoledronate (1-10 mg/kg); (3) a single nephrotoxic dose of the non-nitrogen-containing bisphosphonate, clodronate (2 x 200 mg/kg intraperitoneal injection); (4) a single low dose of ibandronate (1 mg/kg); (5) a single high dose of zoledronate (10 mg/kg). Clinical biochemistry and kidney histopathology were performed 1 and/or 4 days after bisphosphonate dosing. The proximal convoluted tubules were the primary target for renal injury. Tubular degeneration and single cell necrosis of the these tubules were observed with all three bisphosphonates on the fourth, but not the first day after dosing. Differences between the bisphosphonates in the type and/or localisation of the lesions were apparent. Granular deposits in the lumen of distal tubules were apparent with the highest dose of zoledronate (10 mg/kg). However, intraluminal debris was proteinaceous with no evidence of any precipitation of bisphosphonate, or formation of aggregates or complexes in the kidney. Generally, biochemical parameters of renal safety and urinary enzymes did not differ significantly from controls. In summary, bisphosphonate-related renal changes did not appear to be due to the precipitation, aggregation or complexation of bisphosphonate, and biochemical parameters of renal safety did not reliably detect this renal injury. PMID- 16364054 TI - In vitro comparison of clodronate, pamidronate and zoledronic acid effects on rat osteoclasts and human stem cell-derived osteoblasts. AB - In the present study we compared the first generation non-nitrogen-containing bisphosphonate, clodronate with second and third generation nitrogen-containing bisphosphonates, pamidronate and zoledronic acid in dynamic rat osteoclast resorption and apoptosis assays and in human mesenchymal stem cell-derived osteoblast assay. We found that due to high bisphosphonate-bone binding affinity, bone surface exposure to clodronate for 3 min. had maximal resorption inhibition. The mechanism of action of both clodronate and zoledronic acid involved osteoclast apoptosis, whereas pamidronate had only minor apoptotic effect at dosages, which readily inhibited resorption. Zoledronic acid was not metabolised into an intracellular ATP-analogue in vitro in contrast to clodronate. All bisphosphonates had a dose-dependent inhibitory effect on the human bone marrow mesenchymal stem cell (hMSC)-derived osteoblast calcium deposition. None of the compounds had inhibitory effect on hMSC differentiation. Zoledronic acid was the most potent of all three bisphosphonates in terms of both apoptosis induction and resorption inhibition. Zoledronic acid efficacy might thus use its capacity to trigger osteoclast apoptosis in an unknown, but similar manner to that of the non nitrogen-containing bisphosphonates. It appears that zoledronic acid has properties of both bisphosphonate classes and could well be the first member of a new class of bisphosphonates, by definition. PMID- 16364056 TI - Effect of itraconazole on the pharmacokinetics of atenolol. AB - Our objective was to evaluate the effect of itraconazole on the pharmacokinetics of atenolol in healthy volunteers. In a randomized cross-over study with two phases, 10 healthy volunteers had 200 mg itraconazole orally or placebo for 2 days b.i.d., and in the morning of day 3, one hour after the last ingestion of itraconazole or placebo, each subject received 50 mg atenolol. The plasma concentrations of atenolol and its excretion into urine were measured up to 33 hr. Blood pressures and heart rate were recorded up to 10 hr. Itraconazole had no statistically significant effect on any of the pharmacokinetic or pharmacodynamic variables of atenolol. If anything, itraconazole increased the area under the plasma concentration-time curve (+12%; P = 0.159), peak plasma concentration (+19%; P = 0.165), and amount of atenolol excreted into urine (+13%; P = 0.166) suggesting a slight increase of atenolol bioavailability. It can be concluded that itraconazole does not have a clinically relevant effect on the pharmacokinetics of atenolol. PMID- 16364055 TI - Prooxidant action of hinokitiol: hinokitiol-iron dependent generation of reactive oxygen species. AB - Hinokitiol (alpha-thujaplicin, 2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien-1 one), one of the tropolone compounds purified from the woods of Chamaecyparis and Thujopsis (hinoki and hiba), produced reactive oxygen species as a complex with transition metals. Hinokitiol/iron complex inactivated aconitase, the most sensitive enzyme to reactive oxygen, whereas it did not affect aldolase and glyceraldehyde 3-phosphate dehydrogenase. The inactivation of aconitase was iron dependent, and prevented by TEMPOL, a scavenger of reactive oxygen species and superoxide dismutase, suggesting that the hinokitiol/iron-mediated generation of superoxide anion is responsible for the inactivation of aconitase. Addition of hinokitiol effectively enhanced the ascorbate/copper-mediated formation of 8 hydroxy-2'-deoxyguanosine in DNA. Cytotoxic effect of hinokitiol can be explained by its prooxidant properties: hinokitiol/transition metal complex generates reactive oxygen species causing inactivation of aconitase and production of hydroxyl radical resulting in the formation of DNA base adduct. PMID- 16364057 TI - 36 kDa glycoprotein isolated from Rhus verniciflua stokes inhibits G/GO-induced mitochondrial apoptotic signal pathways in BNL CL.2 cells. AB - Rhus verniciflua Stokes is one of the medicinal plants traditionally used to heal and treat hepatic and inflammatory diseases. We found that a glycoprotein isolated from the fruit has a molecular weight of 36 kDa and consists of a carbohydrate component (38.75%) and a protein (61.25%), and that the glycoprotein has a strong scavenging activity against hydroxyl radicals without any pro oxidant activity in the cell-free system. In glucose/glucose oxidase (G/GO) induced BNL CL.2 cells, the results showed that Rhus verniciflua Stokes glycoprotein has dose-dependent blocking activities against G/GO-induced cytotoxicity and apoptosis, increasing the glutathione (GSH) peroxidase activity. In the activity of the mitochondrial apoptotic mediators (cytochrome c, caspases and poly(ADP-ribose)polymerase (PARP)), the glycoprotein (100 microg/ml) showed an inhibitory effect on cytochrome c release, caspase-9/3 activation, and PARP cleavage. Moreover, Rhus verniciflua Stokes glycoprotein has a stimulating effect on the nitric oxide production. Here, we speculate that this glycoprotein is one of the natural antioxidants and of the modulators of apoptotic signal pathways in BNL CL.2 cells. PMID- 16364058 TI - Anterior cingulate activity correlates with blood pressure during stress. AB - The anterior cingulate cortex presumptively regulates blood pressure reactions to behavioral stressors. There is little evidence in humans, however, that stressor evoked changes in blood pressure correlate with concurrent changes in anterior cingulate activity. Using fMRI, we tested whether changes in mean arterial blood pressure correlate with ongoing changes in blood oxygen level dependent (BOLD) activation in 9 women and 11 men who completed a stressful Stroop color-word interference task. Higher mean arterial pressure during the Stroop task correlated with greater BOLD activation in two regions of the cingulate cortex (perigenual and mid-anterior) and in other networked brain regions, including the insula, thalamus, and periaqueductal gray. These results support the hypothesis that the anterior cingulate cortex regulates blood pressure reactions to behavioral stressors in humans. PMID- 16364059 TI - fMRI in an oddball task: effects of target-to-target interval. AB - The amplitude of the P3 event-related potential (ERP) elicited by task-relevant target ("oddball") stimuli has been shown to vary in proportion to the length of time between targets. Here we use functional magnetic resonance imaging (fMRI) to identify neural systems modulated by target interval in a large sample of healthy adults (n=100) during performance of an auditory oddball task that included both target and novel stimuli. A positive relationship was found between target interval and hemodynamic activity in the anterior cingulate and in bilateral lateral prefrontal cortex, temporal-parietal junction, postcentral gryi, thalamus, and cerebellum. This modulation likely represents updating of the working memory template for the target stimuli. There was no such effect of novel interval, suggesting that neuronal modulation may only occur for task-relevant stimuli, possibly in the service of strategic resource allocation processes. PMID- 16364060 TI - Long-term memory is the representational basis for semantic verbal short-term memory. AB - The present study supports activation models of verbal short-term memory that include a semantic contribution to the retention process. Event-related brain potentials were used to probe the level of activation of semantic representations of a series of words in a delay interval following their presentation. The levels of activation were compared in two tasks: (1) a short-term memory task that involved a semantic judgment in the recall phase following the delay interval, and (2) a nonmemory control task. The level of semantic activation during the delay interval was higher in the short-term memory task, indicating that enhanced activation of semantic representations is involved in the short-term storage of verbal information. This result implies that activated long-term memory provides a representational basis for semantic verbal short-term memory, and hence supports theories that postulate that short- and long-term stores are not separate. PMID- 16364061 TI - Meaningful gestures: electrophysiological indices of iconic gesture comprehension. AB - To assess semantic processing of iconic gestures, EEG (29 scalp sites) was recorded as adults watched cartoon segments paired with soundless videos of congruous and incongruous gestures followed by probe words. Event-related potentials time-locked to the onset of gestures and probe words were measured in two experiments. In Experiment 1, participants judged the congruency between gestures and cartoons. Gestures elicited an N400-like component (gesture N450) that was larger for incongruent than congruent items, as well as a late positivity that was larger for congruent items. In Experiment 2, participants assessed the relatedness between probe words and preceding cartoon-gesture pairs. N450 effects to gestures were observed without overlapping positivity. These findings suggest that iconic gestures are subject to semantic processes analogous to those evoked by other meaningful representations, such as pictures and words. PMID- 16364062 TI - Auditory streaming affects the processing of successive deviant and standard sounds. AB - This study investigated the temporal relation between two early mechanisms of auditory information processing: the segregation of the auditory input into streams and the automatic deviance detection indicated by the mismatch negativity (MMN). To address this question the processing of successive deviant and standard tones within streaming and nonstreaming conditions was analyzed. In the streaming condition the amplitude reduction of MMN elicited by the second of two successive deviants was found to be smaller for successive deviants presented in different than in same streams. No corresponding MMN differences were obtained in a nonstreaming condition. These results demonstrate that stream segregation precedes deviance detection. Moreover, modulations of the N1 amplitudes elicited by successive standard tones in the streaming condition demonstrate that not only deviance-related processing but even initial sound processing is affected by streaming. PMID- 16364063 TI - An ERP indicator of processing relevant gestalts in masked priming. AB - Briefly presented arrows, made indistinguishable by masks that contain arrows, inversely prime responses to following visible arrows. This inverse effect might reflect general regularities of masked priming or be either due to the task relevant elements of the mask or to special features of arrows. Here we report a slow negative EEG potential recorded from the scalp above the visual cortex, which is evoked by masks that contain arrows. Even being evoked when arrows masks were presented in isolation, this "Nd-mask" appeared to be an obligatory response. Yet Nd-mask was enhanced when primes and targets were arrows and was reduced in the other cases, and even reversed its polarity with appropriate control stimuli. These findings provide support both for the special status of arrows and for the notion of mask relevance. Nd-mask might be one instance of negative EEG potentials evoked by stimuli with familiar gestalts. PMID- 16364064 TI - Heritability of background EEG across the power spectrum. AB - We estimated the genetic and nongenetic (environmental) contributions to individual differences in the background EEG power spectrum in two age cohorts with mean ages of 26.2 and 49.4 years. Nineteen-lead EEG was recorded with eyes closed from 142 monozygotic and 167 dizygotic twin pairs and their siblings, totaling 760 subjects. We obtained power spectra in 24 bins of 1 Hz ranging from 1.0 to 25.0 Hz. Generally, heritability was highest around the alpha peak frequency and lower in the theta and delta bands. In the beta band heritability gradually decreased with increasing frequency, especially in the temporal regions. Genetic correlations between power in the classical broad bands indicated that half to three-quarters of the genetic variance can be attributed to a common source. We conclude that across the scalp and most of the frequency spectrum, individual differences in adult EEG are largely determined by genetic factors. PMID- 16364065 TI - Effects of aging and working memory demands on prospective memory. AB - The current study used event-related brain potentials (ERPs) to examine the effects of aging, increasing the working memory demands of the ongoing activity, and a prospective memory load on the neural correlates of prospective remembering and target recognition. The behavioral data revealed that the success of prospective memory was sensitive to working memory load in younger, but not older, adults and that a prospective memory load had a greater effect on the performance of older adults than that of younger adults. The ERP data revealed age-related differences in the neural correlates of the detection of prospective cues, post-retrieval processes that support prospective memory, and target recognition. Our results support the hypothesis that there are age-related differences in the ability to recruit preparatory attentional processes that underlie prospective memory, and demonstrate that younger and older adults may recruit somewhat different neural generators to support prospective memory and working memory. PMID- 16364066 TI - Effects of variation in posture and respiration on RSA and pre-ejection period. AB - The extent to which variation in posture and respiration can confound pre ejection period and respiratory sinus arrhythmia (RSA) as indices of cardiac sympatho-vagal activity was examined. Within-subjects changes in these measures were assessed in 36 subjects during different postures and (paced) respiratory frequencies. Changes from supine to sitting to standing led to reduced RSA values and longer pre-ejection periods, reflecting the known decrease in vagal but not the increase of sympathetic activity. Multilevel path analysis showed that within subjects changes in sympatho-vagal balance were faithfully reflected by changes in interbeat interval, but imperfectly by changes in RSA and pre-ejection period. It was concluded that pre-ejection period should be stratified for posture and RSA for respiratory frequency to reliably index changes in sympatho-vagal balance when these factors are prone to change (e.g., during 24-h ambulatory recording). PMID- 16364067 TI - Gender and suppression of mid-latency ERP components during stress. AB - Substantial research evidence suggests that women may be more reactive to stress than men. This study examined the influence of gender and stress on suppression of the P50 and N100 components of the auditory event-related potential. During a stressor task, women (n=13) showed disrupted P50 and N100 suppression whereas men (n=15) exhibited only alterations in N100 suppression. Additionally, reduced skin conductance level during stress correlated with impaired P50 suppression and elevated Click 2 amplitude of the P50 response in women. These data suggest that gender differences in response to perceived stress may be an important factor to consider in studies relying upon the P50 suppression paradigm. PMID- 16364068 TI - Modulation of spinal reflexes by sexual films of increasing intensity. AB - Sexual arousal can be viewed as an emotional state generating sex-specific autonomic and general somatic motor system responses that prepare for sexual action. In the present study modulation of spinal tendious (T) reflexes by sexual films of varying intensity was investigated. T reflexes were expected to increase as a function of increased film intensity. Through use of a between-subjects design, participants were exposed to three erotic films of low, moderate, and high intensity or to three films of moderate intensity. Self-report and genital data confirmed the induction of increasing versus stable levels of sexual arousal. Exposure to the films of increasing intensity resulted in increasing T reflexes. The results indicate that T reflex modulation is sensitive to varying levels of sexual arousal and may be of use in research on behavioral mechanisms underlying appetitive motivation. PMID- 16364069 TI - Differentiation between protective reflexes: cardiac defense and startle. AB - Rise time and duration are two parametric characteristics of the eliciting stimulus frequently used to differentiate among psychophysiological reflexes. The present research varied the duration (study 1) and rise time (study 2) of an intense acoustic stimulus to dissociate cardiac defense and cardiac startle using the eyeblink response as the external criterion of startle. In each study, 100 participants were presented with five white noise stimuli of 105 dB under one of five duration (50, 100, 250, 500, and 1000 ms) or rise time (0, 24, 48, 96, and 240 ms) conditions. Cardiac defense was affected by stimulus duration, present only in the 500- and 1000-ms conditions, but not by stimulus rise time, present in all rise time conditions. Rise time affected blink startle, but did not selectively alter the short latency accelerative component of the heart rate response, thus questioning whether it reflects startle. PMID- 16364070 TI - The latent state-trait structure of resting EEG asymmetry: replication and extension. AB - Recent research on brain asymmetry suggested that resting electroencephalographic (EEG) asymmetry represents a superimposition of a trait-like asymmetry with state dependent fluctuations. The present study tested this hypothesis and additionally examined individual differences in state changes. A 61-channel EEG was collected from 59 participants in a resting state on three occasions of measurement. An analysis of latent state-trait models suggested that between 40% and 50% of the variance of anterior asymmetry measures was due to individual differences on a latent trait and approximately the same portion of the variance was due to occasion-specific fluctuations. A further analysis of true intraindividual change models indicated that there were large individual differences in intraindividual change over time. These data replicate previous findings and substantiate that resting asymmetry has trait and state properties. PMID- 16364075 TI - Hepatitis B virus (HBV) DNA levels and the management of HBV-infected health care workers. AB - Different guidelines exist for the management of hepatitis B virus (HBV)-infected health care workers (HCWs). Various HBV DNA levels are used as a cutoff level to determine whether an HBV-infected HCW is allowed to perform exposure-prone procedures (EPPs) or not. In this paper we discuss the factors that determine HBV DNA levels and the implications of different HBV DNA cutoff levels for EPP performing HCWs. If the level of HBV DNA in the serum of HCWs is used to determine acceptability for the conduct of EPPs, it is necessary to take into account the variability in time of HBV DNA levels in HBV carriers and the reliability and reproducibility of the molecular diagnostic test involved. The issue of standardization has to be addressed, before a universal, maximum level of viraemia for EPP performing HCWs can be introduced. PMID- 16364071 TI - Psychopathy, startle blink modulation, and electrodermal reactivity in twin men. AB - Psychopathy is a personality disorder with interpersonal-emotional and antisocial deviance facets. This study investigated these facets of psychopathy prospectively using normal-range personality traits in a community sample of young adult men who completed a picture-viewing task that included startle blink and skin conductance measures, like tasks used to study psychopathy in incarcerated men. Consistent with prior research, scores on the interpersonal emotional facet of psychopathy ("fearless dominance") were associated with deficient fear-potentiated startle. Conversely, scores on the social deviance facet of psychopathy ("impulsive antisociality") were associated with smaller overall skin conductance magnitudes. Participants high in fearless dominance also exhibited deficient skin conductance magnitudes specifically to aversive pictures. Findings encourage further investigation of psychopathy and its etiology in community samples. PMID- 16364076 TI - Combination hepatitis C virus antigen and antibody immunoassay as a new tool for early diagnosis of infection. AB - Reduction of the window period of hepatitis C virus (HCV) infection represents an important goal in the transfusional and diagnostic setting. A prototype assay designed to simultaneously detect circulating HCV antigen and anti-HCV, has been developed. Aim of this study was to evaluate the performance of this new assay in terms of specificity and sensitivity and to compare its efficacy with commercial assays. To evaluate the specificity of the assay, 400 samples from the general population and 100 'difficult' sera, negative for anti-HCV, were tested. To assess sensitivity, the new test was used on 76 PCR-positive and anti-HCV negative sera, seven natural or commercial seroconversion panels that included 17 RNA-positive and anti-HCV negative sera and 31 anti-HCV positive sera, 20 weak anti-HCV positive sera, 80 viraemic and anti-HCV-positive sera from patients infected with different subtypes and 10 sera from patients with HBV-HCV or HIV HCV co-infections. Of 500 anti-HCV negative samples, 499 (99.8%) were negative with a cut-off index <0.5, while one sample was within the grey zone. Of the 93 HCV-RNA positive and anti-HCV negative sera from patients and panels, 85 (91.4%) resulted positive, and one had the cut-off index in the grey zone. The reduction in the diagnostic window period observed with the new test and HCV-RNA assays were equal, on average, to 24 and 34.4 days respectively. All anti-HCV positive sera were positive. The new assay shows high sensitivity and specificity and could be a useful tool not only in the diagnostic setting, where procedures to reduce the window period, such as antigen or HCV-RNA detection, are not currently recommended, but also in the screening of blood donations, when nucleic acid technologies is not feasible because of costs, organization, emergency and/or logistic difficulties. PMID- 16364077 TI - Interaction between desialylated hepatitis B virus and asialoglycoprotein receptor on hepatocytes may be indispensable for viral binding and entry. AB - The cellular receptor for hepatitis B virus (HBV) infection has not yet been identified. The purpose of this study was to address the possibility of participation by desialylated HBV and the asialoglycoprotein receptor (ASGP-R) exclusively expressed on liver parenchymal cells, in infection. Assays for viral binding and entry were performed by culturing a hepatoblastoma cell line, HepG2, and HBV particles derived from the HBV carrier in the presence or absence of neuraminidase (NA). Viral binding and entry were clearly enhanced in the presence of NA, and the enhancement of the binding could be blocked by asialo-fetuin and ethylenediamine-tetraacetic acid (EDTA). In addition, covalently closed circular (CCC)-DNA, as a marker of infectivity, was detected in the presence of NA, but not in its absence. The optimal concentration of NA raised infectivity more than 1000 times. We concluded that this method makes it feasible to evaluate the infectivity of HBV in vitro and that ASGP-R may be a specific HBV receptor once viral particles are desialylated. PMID- 16364078 TI - Molecular epidemiology of hepatitis C virus (HCV) in Greece: temporal trends in HCV genotype-specific incidence and molecular characterization of genotype 4 isolates. AB - This study aimed to estimate the overall HCV genotype distribution and to reconstruct the HCV genotype-specific incidence in Greece during the recent decades. It also focused at the identification of genotype 4 subtype variability in Greek isolates. A total of 1686 chronically infected HCV patients with detectable serum HCV RNA by RT-PCR, belonging to different risk groups were studied. Amplified products from the 5'-noncoding region were typed using a commercially available assay based on the reverse hybridization principle. The HCV genotype-specific incidence was estimated using a previously described back calculation method. HCV genotype 1 was the most prevalent (46.9%) followed by genotype 3 (28.1%), 4 (13.2%), 2 (6.9%) and 5 (0.4%). A high prevalence of genotype 1 (66.3%) in haemophilia patients was recorded whereas HCV genotype 3 was found mainly among patients infected by I.V. drug use (58.2%). Data on the temporal patterns of HCV genotype-specific incidence in Greece revealed a moderate increase (1.3-1.6 times) for genotypes 1 and 4, and a decrease (1.5 times) for genotype 2 from 1970 to 1990, whereas there was a sharp (13-fold) increase for genotype 3. The molecular characterization of 41 genotype 4 HCV isolates belonging to various risk groups revealed that, subtype 4a was the most frequently detected (78%). Phylogenetic comparison of the Greek 4a isolates with all HCV-4a isolates reported worldwide so far revealed a topology which does not discriminate Greek isolates from the others. HCV-4 does not represent a recent introduction in Greece. PMID- 16364079 TI - Long-term evolution of serum and liver viral markers in patients treated for chronic hepatitis C and sustained response. AB - Few studies have analysed the evolution of HCV markers in chronic hepatitis C (CHC)-treated patients. We have evaluated the presence or absence of serum and liver HCV-RNA, the core antigen (HCV-cAg) and the loss of specific antibodies (anti-HCV), in long-term sustained responders (SR). One hundred and seventy-six patients (132 SR and 44 nonresponders (NR) were included in the study. HCV-RNA was determined in serum and liver by a commercial PCR-kit. HCV-Ag was determined by ELISA and specific antibodies against HCV by means of a commercial line immunoblot assay (LIA) technique. Serum HCVcAg was found positive in three (4.2%) SR and in one (4%) NR (NS). Four SR (3.6%) and 44 NR (100%) were also HCV-RNA (+) in liver tissue. Two patients were HCV-cAg (+). A good correlation was found between the serum levels of HCV-cAg and HCV-RNA (r = 0.847, P < 0.001). Specific antibodies (anti-HCV) were determined by LIA in 45 patients. A decrease was found in the number of patients who presented reactivity to bands E2 and NS4 when we compared SR with a follow-up of more than 5 years with NR and SR with a follow-up <5 years (P < 0.01 and 0.005). A good correlation was found between the HCV-cAg and HCV-RNA serum levels in CHC-treated patients (P < 0.001). Few SR (3.6%) had HCV-RNA in the liver, and HCV-cAg (1.8%) in serum. In SR with more than 5 years of follow-up a clear tendency exists in the trend to clarify the bands E2 and NS4 of anti-HCV in serum. PMID- 16364080 TI - Spontaneous viral clearance following acute hepatitis C infection: a systematic review of longitudinal studies. AB - A large number of studies have reported on spontaneous viral clearance rates in acute hepatitis C infection, however most have been small, and reported rates have varied quite widely. To improve the precision of the estimated rate of spontaneous viral clearance, a systematic review was conducted of longitudinal studies. Factors associated with viral clearance were also examined. Inclusion criteria for studies were: longitudinal assessment from time of acute hepatitis C; hepatitis C virus RNA analysis as determinant of viral clearance; untreated for acute hepatitis C. Information on study population, and factors that may influence viral clearance were extracted from each study. Viral clearance was defined among individuals with at least 6 months follow-up following acute hepatitis C. The number of subjects with viral clearance was expressed as a proportion for each study and a weighted mean for proportion was calculated. A total of 31 studies were examined. Study populations included nine studies of post-transfusion hepatitis, 19 of acute clinical hepatitis, and three of sero incident cases. In total, data was available for 675 subjects and the mean study population was 22 (range 4-67). The proportion with viral clearance ranged from 0.0 to 0.8, with a weighted mean of 0.26 (95% CI 0.22-0.29). Factors associated with viral clearance were female gender and acute clinical hepatitis C study population. Further studies are required to more clearly define predictors of clearance and guide therapeutic intervention strategies. PMID- 16364081 TI - The tandem-repeat polymorphism of the DC-SIGNR gene in HCV infection. AB - The C-type lectin DC-SIGNR has been shown to bind hepatitis C virus (HCV). Here, we analysed the tandem-repeat polymorphism of the DC-SIGNR gene with respect to intraindividual HCV replication. In a cross-sectional comparison HCV-infected patients (n = 430) and healthy subjects (n = 100) were genotyped for the DC-SIGNR polymorphism using PCR. The distribution of DC-SIGNR alleles did not differ significantly between the two groups. However, HCV-infected patients with 5-, 6-, and 7-repeat alleles had higher HCV-RNA levels when compared with carriers of 4- and 9-repeat alleles (P < 0.05). Thus, the DC-SIGNR polymorphism might affect HCV loads supporting the concept that DC-SIGNR contributes to HCV replication efficacy. PMID- 16364082 TI - Characterization of T-cell responses against immunodominant epitopes from hepatitis C virus E2 and NS4a proteins. AB - Successful clearance of hepatitis C virus (HCV) infection has been associated with strong cellular immune responses against viral antigens. However, although the magnitude of these responses is clearly important for viral eradication, more studies are needed to unravel the fine specificity of the protective anti-HCV immunity in infected patients. This was the aim of the present study. Overlapping peptides spanning the sequence of HCV E2 and NS4a proteins were used to stimulate T cells from patients with chronic hepatitis C divided into three groups: naive patients, patients who exhibited sustained response to interferon (IFN)-alpha therapy and patients who failed to respond to the treatment. Interleukin-2 production by stimulated cells was measured in each case. Patients with sustained response to therapy had stronger responses to E2 peptides than nonresponders, whereas naive patients demonstrated intermediate reactivity. In the case of NS4a, responses against peptides where similar in all groups of patients. Analysis of the peptides recognized by T cells showed that responses were broad and heterogeneous, and some immunodominant epitopes, preferentially recognized by patients exhibiting sustained response to treatment, were found. These results confirm the role of cellular immune responses in viral clearance, and stress the importance of immunodominant regions within HCV antigens. These viral sequences may represent valuable immunogens for preparation of therapeutic or prophylactic vaccines. PMID- 16364083 TI - Serum lipid pattern in chronic hepatitis C: histological and virological correlations. AB - Lipoproteins are closely connected to the process of hepatitis C virus (HCV) infection. The aim of this study was to evaluate the lipaemic profile in patients with chronic HCV infection, and to identify any association between serum lipid levels and viral load, HCV genotype or liver histology. Total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C) and triglycerides (TG) were measured in the sera of 155 patients with chronic HCV infection and 138 normal subjects, matched for age and sex. Viral parameters and liver histology were evaluated in HCV-infected patients. Serum TC (P < 0.0005), HDL-C (P < 0.0005) and LDL-C (P < 0.0005) were lower in chronic hepatitis C patients compared with controls. Grading score was positively correlated with TC and LDL-C. Patients with HCV genotype 3a had significantly lower levels of TC, HDL-C, LDL-C, higher viral load and higher frequency of hepatic steatosis than those with other genotypes. Logistic regression analysis identified genotype 3a (OR, 6.96; 95% CI, 2.17-22.32, P = 0.0011) as the only significant predictive variable associated with low serum cholesterol concentration. HCV infection is associated with clinically significant lower cholesterol levels (TC, LDL and HDL) when compared with those of normal subjects. This finding is more pronounced in patients infected with HCV genotype 3a. Further studies are necessary to define the pathophysiology of the relationship between lipid metabolism and HCV infection. PMID- 16364084 TI - Characterization of hepatitis B surface antigen strains circulating in the Kingdom of Cambodia. AB - A collection of hepatitis B surface antigen (HBsAg)-reactive serum specimens from the Pasteur Institute of Cambodia was investigated for the genotype, predicted serotype and the presence of diagnostically significant mutations in the surface protein. From a set of 794 samples, 15 were identified serologically to harbour possible HBsAg mutants and were investigated further. An additional 20 samples were included into the study for PCR and sequence analysis. Of the 22 samples which were HBV-DNA-positive, 16 were of genotype C with the remaining six being genotype B. Point mutations resulting in amino acid substitutions were noted in 10 samples. The majority of these mutations occurred outside the a determinant. PMID- 16364085 TI - Exposure to HAV infection in patients with chronic liver disease in Italy, a multicentre study. AB - We carried out a multicentre study on 2830 patients with chronic liver disease from 79 liver units (25 in northern, 24 in central and 30 in southern Italy) to evaluate naturally acquired immunity against hepatitis A virus (HAV) in relation to age, sex, geographical area of origin and entity of liver disease, and to define the strategy for specific vaccination. Antibody to HAV (anti-HAV) was detected in 1514 (53.5%) of the 2830 patients tested; the prevalence was 50.4% in males and 59.1% in females. Both in central and southern Italy the prevalence of anti-HAV positive subjects increased with increasing age from 43.3 and 44.7%, respectively, in the 0-30-year-old subjects to 80.1 and 68.3%, respectively, in those aged over 60 years. The overall prevalence was much lower in northern Italy, as were the variations from one age group to another, from 28.4% in the 0 30-year-old subjects to 38% in those aged over 60 years. 40.6% of patients with cirrhosis lacked naturally acquired protection against HAV; this percentage was higher in northern (60.5%) than in central (34.9%, P < 0.0001) and southern Italy (27.6%, P < 0.0001). The high prevalence of patients in Italy with chronic hepatitis or cirrhosis who lack naturally acquired immunity to HAV warrants the implementation of vaccination programmes against hepatitis A in such patients. PMID- 16364087 TI - Immunocytochemical investigation of immune cells within human primary and permanent tooth pulp. AB - AIM: The aim of this study was to determine whether there are any differences in the number and distribution of immune cells within human primary and permanent tooth pulp, both in health and disease. DESIGN: The research took the form of a quantitative immunocytochemical study. One hundred and twenty-four mandibular first permanent molars and second primary molars were obtained from children requiring dental extractions under general anaesthesia. Following exodontia, 10 microm-thick frozen pulp sections were processed for indirect immunofluorescence. Triple-labelling regimes were employed using combinations of the following: (1) protein gene product 9.5, a general neuronal marker; (2) leucocyte common antigen (LCA); and (3) Ulex europaeus I lectin, a marker of vascular endothelium. Image analysis was then used to determine the percentage area of immunostaining for LCA. RESULTS: Leucocytes were significantly more abundant in the pulp horn and mid-coronal region of intact and carious primary teeth, as compared to permanent teeth (P < 0.05, anova). Both dentitions demonstrated the presence of well localized inflammatory cell infiltrates and marked aborization of pulpal nerves in areas of dense leucocyte accumulation. CONCLUSIONS: Primary and permanent tooth pulps appear to have a similar potential to mount inflammatory responses to gross caries The management of the compromised primary tooth pulp needs to be reappraised in the light of these findings. PMID- 16364088 TI - Examining intention to control preschool children's sugar snacking: a study of carers in Uganda. AB - AIM: The aims of this study were to investigate parents' intention to control their children's sugar consumption and whether that behaviour is reported to occur in 3-5-year-old preschool children according to sociodemographics and attitudinal factors derived from the Theory of Planned Behaviour (TPB). DESIGN: Some 589 children aged 3-5 years (51% boys, response rate = 85%) attending nursery schools in Kampala Central (urban) and Nakawa (suburban), Uganda, were examined clinically for dental caries. A questionnaire to assess sociodemographic factors, sugar intake and the constructs of the TPB was completed by their parents'/caregivers in face-to-face interviews. RESULTS: Analyses of variance revealed more positive attitudes and stronger intention to control children's intake of sugared snacks in highly as compared to less highly educated parents. Independent of educational level, parents having children with caries perceived themselves to have less control over their child's intake of sugared snacks and perceived them to be more susceptible to tooth decay compared to parents of children without caries. In multiple linear regression, the TPB provided a significant prediction of intention with attitude (b = 0.16, P < 0.001), subjective norms (b = 0.18, P < 0.001) and perceived barriers (b = 0.11, P = 0.01), significant and reported sugar intake with attitudes (b = -0.10, P = 0.02), and perceived susceptibility (b = 16, P < 0.001) all significant. CONCLUSION: The TPB components predict parental intention to control sugar snacking and whether that behaviour occurs in preschool children. The strengths of parents' attitudes and reported level of child sugar snacking varied between diverse socioeconomic family groups. Implications for oral health education are discussed. PMID- 16364089 TI - An analysis of oral and maxillofacial pathology found in children over a 30-year period. AB - AIM: The vast majority of oral diseases are confined to oral tissues, but numerous underlying systemic conditions may present with signs and symptoms within the oral cavity. Since the epidemiology of diseases is variable between regions, the authors carried out Europe's first paediatric-based survey of oral and maxillofacial pathology specimens submitted for diagnosis. DESIGN: All entries for specimens from children between the ages of 0 and 16 years during the 30-year period from 1973 to 2002 were retrieved and compiled into 12 diagnostic categories. RESULTS: During the study period, 4406 (8.2%) specimens came from children between the ages of 0 and 16 years, with a male to female ratio of 1.01. The diagnostic category with the largest number of specimens was tooth pathology (22.1%), followed by salivary gland disease (19.1%) and mucosal pathology (12.1%). In all, there were 114 benign tumours of nonodontogenic origin, 43 odontogenic tumours and 31 malignant tumours. The most frequently diagnosed lesions were mucous extravasation cysts, which accounted for over 16% of cases. Periapical pathology in the form of a radicular cyst, residual cyst or chronic periapical granuloma formed almost 13% of all cases. CONCLUSIONS: This survey shows that, while nearly 10% of specimens submitted to the authors' laboratory are from children under 16 years of age, the majority of lesions are of a benign nature, requiring minimal intervention; less than 1% of cases comprise malignant lesions. Odontogenic tumours are relatively rare in this age group; however, certain lesions such as adenomatoid odontogenic tumour and ameloblastic fibroma occur predominantly in children and, therefore, remain an important diagnostic consideration. PMID- 16364090 TI - Differences in methodologies of measuring the prevalence of oral mucosal lesions in children and adolescents. AB - AIM: This review offers a brief summary of three of the more commonly surveyed and reported oral mucosal conditions found in children, and an appraisal of the variation between selected elements of prevalence study methodologies. DESIGN: Searches of electronic databases (i.e. MEDLINE, CINAHL, EMBASE and EBM Reviews) were conducted to find publications related to oral mucosal conditions in children. Studies were included and reviewed if they contained data on the prevalence of any of the following oral mucosal conditions: geographical tongue, oral ulceration and herpes labialis in children or adolescents (up to the age of 19 years). RESULTS: A total of 29 articles out of 333 met the inclusion criteria and were reviewed. Of these 29 reports, 18 considered geographical tongue, 12 herpes labialis and 10 oral ulceration, although a number of the surveys reported on more than one condition. The approaches used in these studies varied on a number of the potentially important areas associated with validity, comparability, and whether or not the findings may be extrapolated beyond the study population, such as the sampling frame, diagnostic criteria, training and calibration, and examination features. CONCLUSION: Although there were a number of surveys relating to the prevalence of geographical tongue, herpes labialis and oral ulcers in children and adolescents, reported prevalences varied and few studies were directly comparable in terms of the methodology applied. In particular, there was substantial variation between surveys in terms of the diagnostic criteria and method of detection employed although, in many instances, there was inadequate detail to allow full appraisal of the methodology. There is a need for more good-quality epidemiological studies in this area. PMID- 16364091 TI - An oral health education programme based on the National Curriculum. AB - AIM: The aim of this study was to develop and evaluate a teaching programme based on the national curriculum for use in a primary school setting. DESIGN: National Curriculum guidelines were combined with oral health education messages to draw up lesson plans for teachers to deliver. A questionnaire was used to demonstrate children's oral health knowledge prior to the teaching programme, and at 1 and 7 weeks following the programme. The study took place in inner-city, state-run primary schools in Manchester and North London, UK. The subjects were children between the ages of 7 and 8 years from Manchester (n = 58) and North London (n = 30). The main outcome measure was change in knowledge attributable to a newly developed teaching programme. RESULTS: The children in Manchester had a higher level of knowledge prior to the teaching programme. Following the teaching programme, children in both schools showed a significant improvement in dental health knowledge (P < 0.001). Seven weeks later, the Manchester children showed no significant loss of knowledge (P < 0.001). CONCLUSIONS: The aims of the National Curriculum were easily integrated with oral health messages. A more widely available teaching resource, such as the one described in this study, would be useful to encourage the teaching profession to take on oral health education without more costly input from dental professionals. PMID- 16364092 TI - The frequency of repeat general anaesthesia for teeth extractions in children. AB - AIM: The purpose of this study was to determine the frequency of repeat extractions under general anaesthesia (GA) in children. METHODS: The dental hospital records of patients attending for outpatient GA extractions at Liverpool University Dental Hospital, Liverpool, UK, between January and March 2003 were examined retrospectively. A data collection form was used to record the relevant information. RESULTS: A total of 278 patients with a mean age of 6.5 years (SD = 2.2 years) were seen for GA extractions. Of those, 33 patients (11.9%) with a mean age of 4.9 years (SD = 2 years) at the time of their initial GA had had a previous or would undergo a subsequent episode of GA extraction. The mean interval between repeat GA was 2.3 years (SD = 1.6 years). Fifteen cases (45.5%) had the repeat GA within 2 years. Radiographs were available as part of the assessment process for 84 (34.3%) of the 245 patients who had had a single episode of GA. However, of the 33 patients who had had a repeat GA, only seven (21.2%) had radiographs available at the time of the initial GA. Regarding the number of teeth extracted, a significant difference (P < 0.01) was found between the number of teeth extracted in patients who had had a single GA (mean = 4.6, SD = 2.5), compared with those extracted at the initial GA for the repeat GA group (mean = 3.2, SD = 2). CONCLUSION: The frequency of repeat GA is relatively low, but there is a need for appropriate treatment planning incorporating the use of radiographs to reduce this even further. PMID- 16364093 TI - Clinical performance of resin-bonded composite strip crowns in primary incisors: a retrospective study. AB - AIM: The purpose of this study was to assess retrospectively the longevity of resin-bonded composite strip crowns placed in primary maxillary incisors. DESIGN: Records for 200 out of 387 children, aged 22-48 months, treated in a private paediatric dental practice and who presented for follow-up after at least 24 months were included in the study. The parameters recorded at baseline and/or at follow-up were: habits, the number and location of the decayed surfaces, colour, texture, and chipping of the restoration. Radiographic evaluation of the restorations, the quality of the margins, and the presence of pulpal and/or periapical pathoses were recorded. RESULTS: More than 80% of the restorations were judged to be successful at the final follow-up examination. Only the number of carious surfaces of the tooth at baseline influenced the treatment outcome. The failure rate was higher in central incisors with four affected surfaces (P = 0.005), and in lateral incisors with four carious surfaces (P = 0.0003), than in those presenting one or two carious surfaces in both central and lateral incisors (P = 0.002). CONCLUSION: The high success rate of resin-bonded composite strip crowns with a 2-year follow-up seen in this study suggests that this treatment modality is an aesthetic and satisfactory means of restoring carious primary incisors in young children. The retention rate is lower in teeth with decay in three or more surfaces, particularly in children with a high caries risk. PMID- 16364094 TI - An unusual complication of mandibular distraction. AB - BACKGROUND: The authors present an unusual complication of mandibular distraction in a child with the curious condition of multiple pterygium syndrome is presented. CASE REPORT: The patient was a Caucasian male with severe pterygia in his neck. As a result of his limited mouth opening and restricted upper airway leading to obstruction, he underwent lengthening of his mandible by distraction, which significantly improved his breathing. During his follow-up, it was observed that an unusually elongated permanent molar was present in an abnormal position. CONCLUSION: This case highlights the need to carefully plan the sites for osteotomy and the potential for damage to the developing permanent dentition in young children. PMID- 16364095 TI - Periostitis ossificans (Garre's osteomyelitis) radiographic study of two cases. AB - BACKGROUND: Periostitis Ossificans (PO) is a non-suppurative type of Osteomyelitis, commonly occurring in children and young adults, in mandible. The most common cause for PO is periapical infection of mandibular first molar. Radiographically PO is characterized by the presence of lamellae of newly formed periosteal bone outside the cortex, giving the characteristic appearance of "onion skin". CASE REPORTS: Two male children 11 years of age reported to the Department of Oral Medicine with a painless and persistent bony hard swelling in the mandible, with a short duration (Figs 1, 5). Both the patients had grossly decayed mandibular permanent first molar tooth with periapical infection and buccal cortical plate expansion (Figs 2, 6). The radiographic study revealed different appearances, the Orthopantomograph of case I showed a single radiopaque lamella outside the lower cortical border, without altering original mandibular contour (Fig. 3) and in case II showed a newly formed bony enlargement on the outer aspect of the lower cortical border without altering the original mandibular contour (Fig. 7). Occlusal radiograph of both the patients showed two distinct radiopaque lamellae of periosteal bone outside the buccal cortex (Figs 4, 8). Kawai et al. classified PO of mandible into type I and type II, based on whether the original contour of mandible is preserved or not. Each type is further classified into two sub types (Table 1). In case I, the orthopantomographic appearance is characteristic of type I-1 (Fig. 3), but the appearance in occlusal radiograph is characteristic of type I-2 (Fig. 4). In case II, the appearances in both the radiographs are characteristic of type I-2 (Figs 7, 8). CONCLUSIONS: Apart from the typical onion skin appearance, PO shows various other radiographic appearances. The radiographic appearance of Periostitis Ossificans may reflect the duration, progression and the mode of healing of the disease process. The radiographic classification of PO depends on the type of radiographs taken for evaluation. PMID- 16364096 TI - Focal epithelial hyperplasia (Heck's disease): report of a case in a girl of Brazilian Indian descent. AB - Summary. Background. This report describes the case of a patient with focal epithelial hyperplasia (FEH), a rare but distinctive entity of viral aetiology with characteristic clinical and histopathological features. Case report. The condition is usually seen in children and adolescents of American Indian and Eskimo background. Surgical removal of papillomatous lesions is the treatment of choice, either for aesthetic reasons, or when the lesions interfere with function or are readily traumatized. Recurrence and the site of new lesions are unpredictable, and continued review of the patient is often necessary. The patient described here has been followed for 24 months without recurrences or changes in the aspect of the remaining lesions. Conclusion. This case highlights a possible genetic predilection for FEH, since the patient is a descent of a Brazilian Xavante Indian. PMID- 16364097 TI - Noonan syndrome with giant cell lesions. PMID- 16364103 TI - Benefit assessment of preventive medical check-ups in patients suffering from chronic granulomatous disease (CGD). AB - BACKGROUND: Chronic granulomatous disease (CGD) patients are susceptible to recurrent opportunistic infections and to recurrent or chronic inflammations of presumably non-infectious origin. Both types of manifestations are often accompanied by granuloma formation. Both can remain unnoticed until they deteriorate considerably and can become life-threatening if not treated in time. OBJECTIVE: To evaluate the effectiveness of regular follow-up visits in CGD patients. METHODS: Findings of imaging (ultrasound, radiographs, computer tomography, magnetic resonance imaging), lung function tests, histopathological and microbiological assessments of biopsies have been reviewed. RESULTS: A total of 32 CGD patients have been evaluated within 15 years. Of these eight patients have been checked regularly for more than 5 years. Early detection prior to specific clinical signs and consecutive treatment of six lung manifestations and one liver manifestation such as interstitial pneumonia with formation of small granulomas, formation of large granulomas, fibrosis, Aspergillus infections, and abscesses could presumably prevent considerable aggravation in seemingly healthy or mildly symptomatic patients. In contrast, patients without surveillance who presented with severe symptoms (seven manifestations) often had irreversible organ damage or even died. CONCLUSIONS: Regular follow-up visits can help prevent or mitigate clinical manifestations, improve life quality and expectancy and weigh indication for bone marrow transplantation in CGD patients. PMID- 16364104 TI - Audit of the multidisciplinary management of orbital infection secondary to sinusitis. PMID- 16364105 TI - National audit of continence care for older people: results of a pilot study*. AB - INTRODUCTION AND AIM: Bladder and bowel problems are common in the elderly and are associated with a considerable morbidity and impact on quality of life. Inequalities in service provision and access to services have been recognized but there has been no systematic approach to measuring the quality of continence care for older people. This study aimed to develop quality standards, to assess the reliability and utility of the resulting audit package and to report on the standards of care provided in primary care, secondary care and care home setting. METHOD: Fifteen sites in secondary care, primary care and in long-term care settings were randomly selected to pilot the audit package. Data collectors completed audit questionnaires relating to the structure [organization] of care, the outcomes of care, and the process of care for 20 subjects with urinary incontinence and 10 subjects with faecal or double incontinence. RESULTS: The audit tool was reliable (median kappa score of 0.7). Access to integrated continence services, as defined by Good Practice in Continence Services was inadequate. Eighty-five per cent of hospitals had no written policy for continence care. There were deficiencies in obtaining information, in carrying out basic and specialist examinations and investigations and in determining the cause of incontinence. There was a high prevalence of catheter use in secondary care settings. CONCLUSION: The pilot has indicated significant inadequacies in continence care and demonstrates that in many sites the National Service Framework milestone for integrated continence services has not been met. A national audit of continence care is required to determine the extent of inadequate continence care. PMID- 16364106 TI - National audit of continence care: laying the foundation. AB - INTRODUCTION: National audit provides a basis for establishing performance against national standards, benchmarking against other service providers and improving standards of care. For effective audit, clinical indicators are required that are valid, feasible to apply and reliable. This study describes the methods used to develop clinical indicators of continence care in preparation for a national audit. AIM: To describe the methods used to develop and test clinical indicators of continence care with regard to validity, feasibility and reliability. METHOD: A multidisciplinary working group developed clinical indicators that measured the structure, process and outcome of care as well as case-mix variables. Literature searching, consensus workshops and a Delphi process were used to develop the indicators. The indicators were tested in 15 secondary care sites, 15 primary care sites and 15 long-term care settings. RESULTS: The process of development produced indicators that received a high degree of consensus within the Delphi process. Testing of the indicators demonstrated an internal reliability of 0.7 and an external reliability of 0.6. Data collection required significant investment in terms of staff time and training. CONCLUSION: The method used produced indicators that achieved a high degree of acceptance from health care professionals. The reliability of data collection was high for this audit and was similar to the level seen in other successful national audits. Data collection for the indicators was feasible to collect, however, issues of time and staffing were identified as limitations to such data collection. The study has described a systematic method for developing clinical indicators for national audit. The indicators proved robust and reliable in primary and secondary care as well as long-term care settings. PMID- 16364107 TI - Clinical risk: a dangerous situation. PMID- 16364108 TI - Appraising the quality of randomized controlled trials: inter-rater reliability for the OTseeker evidence database. AB - RATIONALE AND AIMS: 'OTseeker' is an online database of randomized controlled trials (RCTs) and systematic reviews relevant to occupational therapy. RCTs are critically appraised and rated for quality using the 'PEDro' scale. We aimed to investigate the inter-rater reliability of the PEDro scale before and after revising rating guidelines. METHODS: In study 1, five raters scored 100 RCTs using the original PEDro scale guidelines. In study 2, two raters scored 40 different RCTs using revised guidelines. All RCTs were randomly selected from the OTseeker database. Reliability was calculated using Kappa and intraclass correlation coefficients [ICC (model 2,1)]. RESULTS: Inter-rater reliability was 'good to excellent' in the first study (Kappas >or= 0.53; ICCs >or= 0.71). After revising the rating guidelines, the reliability levels were equivalent or higher to those previously obtained (Kappas >or= 0.53; ICCs >or= 0.89), except for the item, 'groups similar at baseline', which still had moderate reliability (Kappa = 0.53). In study 2, two PEDro scale items, which had their definitions revised, 'less than 15% dropout' and 'point measures and variability', showed higher reliability. In both studies, the PEDro items with the lowest reliability were 'groups similar at baseline' (Kappas = 0.53), 'less than 15% dropout' (Kappas or =8). 2 propanol-based formulations were tested in a repeated open application test, 1 contained a mixture of emollients (0.81%, w/w). 2 aliquots of 0.7 ml of each formulation were applied twice per day over 2 weeks to the cubital fossa of each subject after random assignment of the preparations. Treatment areas were assessed before each application and 3 days postfinal application by visual inspection for erythema and dryness according to a standard scale. The sum score over all assessment time-points served as primary parameter. The mean sum score for erythema and dryness was significantly lower for the hand rub with emollients (0.8 +/- 2.4) in comparison with that for the hand rub without emollients (1.5 +/ 3.5; P = 0.022; Wilcoxon signed rank test). A comparison of the atopic and non atopic subjects revealed no significant difference for any of the products (P > 0.05; Mann-Whitney U-test). It is concluded that the addition of emollients to a propanol-based hand rub can significantly decrease irritant contact dermatitis under frequent-use conditions. PMID- 16364125 TI - Allergic contact dermatitis from cobalt in jewellery. PMID- 16364126 TI - Contact erythema multiforme from Primula obconica. PMID- 16364127 TI - Delayed-type hypersensitivity to mepivacaine with cross-reaction to lidocaine. PMID- 16364129 TI - Mentha pulegium contact dermatitis. PMID- 16364128 TI - N,N-dimethyl-n-dodecylamine caused contact dermatitis. PMID- 16364130 TI - Allergic contact dermatitis to Plectranthus amboinicus masquerading as chronic leg ulcer. AB - This report discusses a case of a 69-year-old woman who developed chronic non healing leg ulcers after long-term topical use of Plectranthus amboinicus. The ulcer was proven to be allergic contact dermatitis to P. amboinicus by a patch test. The ulcer healed after discontinuation of P. amboinicus. To the best of our knowledge, this is the first reported case of allergic contact dermatitis to P. amboinicus masquerading as chronic leg ulcer. PMID- 16364131 TI - Allergic contact dermatitis due to methyldibromo glutaronitrile in make-up removal wipes. PMID- 16364132 TI - Contact allergy to gold in patients with gold-plated intracoronary stents. PMID- 16364133 TI - Nocturnal ghrelin levels--relationship to sleep EEG, the levels of growth hormone, ACTH and cortisol--and gender differences. AB - Ghrelin, an endogenous ligand of the growth hormone (GH) secretagogue receptor, stimulates sleep, appetite and weight gain as well as the secretion of GH, adrenocorticotropic hormone (ACTH), cortisol in humans and rodents. The interaction between nocturnal ghrelin levels, sleep EEG and the secretion of these hormones was not investigated systematically so far. Furthermore conflicting data exist on gender differences in nocturnal ghrelin secretion. We examined simultaneously sleep EEG and the nocturnal levels of ghrelin, GH, ACTH and cortisol in young and middle-aged normal human subjects (eight males, eight females). A significant interaction between gender and the course of ghrelin concentration was observed to the interval between 20:00 and 23:00 hours. In males a continuous increase of ghrelin levels before sleep onset was found. In females, however, a rise of ghrelin during the night was missed. We found a trend suggesting a lower time spent in stage I sleep in subjects with high nocturnal ghrelin levels. Other systematic interactions between plasma ghrelin, sleep EEG and other hormones were not found. No peak in plasma ghrelin levels resembling the GH surge was observed. We suggest that under naturalistic conditions plasma ghrelin levels show no distinct interaction with sleep. PMID- 16364134 TI - Analysis of oscillatory patterns in the human sleep EEG using a novel detection algorithm. AB - The different brain states during sleep are characterized by the occurrence of distinct oscillatory patterns such as spindles or delta waves. Using a new algorithm to detect oscillatory events in the electroencephalogram (EEG), we studied their properties and changes throughout the night. The present approach was based on the idea that the EEG may be described as a superposition of stochastically driven harmonic oscillators with damping and frequency varying in time. This idea was implemented by fitting autoregressive models to the EEG data. Oscillatory events were detected, whenever the damping of one or more frequencies was below a predefined threshold. Sleep EEG data of eight healthy young males were analyzed (four nights per subject). Oscillatory events occurred mainly in three frequency ranges, which correspond roughly to the classically defined delta (0-4.5 Hz), alpha (8-11.5 Hz) and sigma (11.5-16 Hz) bands. Their incidence showed small intra- but large inter-individual differences, in particular with respect to alpha events. The incidence and frequency of the events was characteristic for sleep stages and non-rapid eye movement (REM)-REM sleep cycles. The mean event frequency of delta and sigma (spindle) events decreased with the deepening of sleep. It was higher in the second half of the night compared with the first one for delta, alpha and sigma oscillations. The algorithm provides a general framework to detect and characterize oscillatory patterns in the EEG and similar signals. PMID- 16364135 TI - Event-related potential measures of the disruptive effects of trains of auditory stimuli during waking and sleeping states. AB - Acoustic backup alarms have been reported to particularly disrupt sleep. The present study simulated backup alarms by presenting trains of five consecutive 500 ms duration audible tones, with the time between the onset of each tone being 1 s and the time between trains (offset to onset) between 15 and 20 s. In different conditions, the tones were set at either 80 or 60 dB sound pressure level (SPL). Twelve young adults spent two consecutive nights in the laboratory. Stimuli were presented only on the second night. Measures of traditional sleep architecture (sleep stages) were not affected by the acoustic trains. Event related potentials were also measured following presentation of the stimuli. In the waking state, the initial 80 dB stimulus elicited a large amplitude N1, peaking at about 100 ms, followed by a positive peak, P3, peaking at about 325 ms. N1 was attenuated following presentation of the 60 dB stimulus. The amplitude of N1 was much reduced following presentation of the subsequent second to fifth stimuli in the train. During non-rapid eye movement (NREM) sleep, the initial 80 dB stimulus elicited a large and later negativity (N350) that was reduced in amplitude for the 60 dB stimulus. A K-Complex (a composite N350 and a much larger N550) was elicited following 35% of the initial 80 dB tones and 12% of the initial 60 dB tones. The amplitude of N550 did not, however, significantly vary as a function of stimulus SPL. During REM sleep, N1 continued to be elicited by the initial louder stimulus, but the later positive wave was not apparent. A late negativity peaking at about 350 ms was, however, apparent. When queried the next morning, subjects rarely indicated that the stimulus presentations disturbed their sleep. This might be because of the absence of the late positivity. The presence of the long latency negativities (N350 and N550) might serve to protect sleep from obtrusive sound during sleep. PMID- 16364136 TI - CAP and arousals are involved in the homeostatic and ultradian sleep processes. AB - There is growing evidence that cyclic alternating pattern (CAP) and arousals are woven into the basic mechanisms of sleep regulation. In the present study, the overnight sleep cycles (SC) of 20 normal subjects were analyzed according to their stage composition, CAP rate, phase A subtypes and arousals. Individual SC were then divided into 10 normalized temporal epochs. CAP parameters and arousals were measured in each epoch and averaged in relation to the SC order. Subtypes A2 and A3 of CAP in non-rapid eye movement (NREM) sleep, and arousals, both in REM and NREM sleep when not coincident with a A2 or A3 phases, were lumped together as fast electroencephalographic (EEG) activities (FA). Subtypes A1 of CAP, characterized by slow EEG activities (SA), were analyzed separately. The time distribution of SA and FA was compared to the mathematical model of normal sleep structure including functions representing the homeostatic process S, the circadian process C, the ultradian process generating NREM/REM cycles and the slow wave activity (SWA) resulting from the interaction between homeostatic and ultradian processes. The relationship between SA and FA and the sleep-model components was evaluated by multiple regression analysis in which SA and FA were considered as dependent variables while the covariates were the process S, process C, SWA, REM-on and REM-off activities and their squared values. Regression was highly significant (P < 0.0001) for both SA and FA. SA were prevalent in the first three SC, and exhibited single or multiple peaks immediately before and in the final part of deep sleep (stages 3 + 4). The peaks of FA were delayed and prevailed during the pre-REM periods of light sleep (stages 1 + 2) and during REM sleep. SA showed an exponential decline across the successive SC, according to the homeostatic process. In contrast, the distribution of FA was not influenced by the order of SC, with periodic peaks of FA occurring before the onset of REM sleep, in accordance with the REM-on switch. The dynamics of CAP and arousals during sleep can be viewed as an intermediate level between cellular activities and macroscale EEG phenomena as they reflect the decay of the homeostatic process and the interaction between REM-off and REM on mechanisms while are slightly influenced by circadian rhythm. PMID- 16364137 TI - Functional brain imaging of a complex navigation task following one night of total sleep deprivation: a preliminary study. AB - Several neuroimaging studies have demonstrated compensatory cerebral responses consequent to sleep deprivation (SD), but all have focused on simple tasks with limited behavioral response options. We assessed the cerebral effects associated with SD during the performance of a complex, open-ended, dual-joystick, 3D navigation task (simulated orbital docking) in a cross-over protocol, with counterbalanced orders of normal sleep (NS) and a single night of total SD (approximately 27 h). Behavioral performance on multiple measures was comparable in the two sleep conditions. Functional magnetic resonance imaging revealed multiple compensatory SD > NS cerebral responses, including the posterior superior temporal sulcus [Brodmann area (BA) 39/22/37], prefrontal cortex (BA 9), lateral temporal cortex (BA 22/21), and right substantia nigra. Right posterior cingulate cortex (BA 31) exhibited NS > SD activity. Our findings extend the compensatory cerebral response hypothesis to complex, open-ended tasks. PMID- 16364138 TI - Investigating couples' sleep: an evaluation of actigraphic analysis techniques. AB - 'Blip' analysis, fast wavelet transformations (FWT) and correlation analysis have all been used to actigraphically assess the impact one person is having on another's sleep, yet no review exists as to the differences between, and applicability of, these methods for investigating couples' sleep. Using actigraphy data and audio sleep diaries collected from 18 couples, this paper provides such a review. This paper constructs and assesses two novel, analytical methods: Lotjonen's sleep/wake algorithm, and the partner impact on sleep wake analysis (PISWA). Both 'blip' analysis and correlation suggest that the strongest relationship between bed partners occurs on an epoch-to-epoch basis. However, 'blips' deal strictly with onset of movement and fail to incorporate strength and duration of movement. Conversely, correlation analysis incorporates some elements of strength and duration of movement but makes identification of onset problematic. FWT offer useful 'relativistic' pattern recognition, identifying onset, strength and duration of movement, but are difficult to quantify. Although audio diary data support the potential of Lotjonen's sleep/wake algorithm to identify sleep non-movement, sleep movement, wake non-movement (or quiet wakefulness) and wake movement, the problem remains that this method also relies on visualization. Of most promise, we argue, is the PISWA, which examines 'impact' of bed partners through incorporating elements of 'blip' analysis and the sleep/wake algorithm. PMID- 16364139 TI - Sleep latency testing as a time course measure of state arousal. AB - The purpose of this study was to determine how long the effects of a brief period of physiological arousal persisted using repeated sleep latency testing and measurement of heart rate. Thirteen normal sleeping young adults spent two non consecutive nights and the following days in the laboratory. On each day, subjects had five sleep latency measurements - at 09:00, 09:30, 10:00, 10:30, and 11:00 hours. The 09:00 test was a premanipulation baseline. Following this nap, subjects either walked for 5 min (on one day) or rested in bed for 10 min (on another day) prior to the 09:30 hours sleep latency test. Significant increases in sleep latency were found at 09:30, 10:00, and 11:00 hours following the single 5-min walk as compared with resting in bed (mean sleep latency after the walk was 11.7 min compared with 7.1 min for the resting condition). Heart rate was significantly higher throughout all of the postmanipulation naps following the walk. The elevated sleep latency is probably secondary to the changes in underlying physiological arousal as measured in this study by heart rate. PMID- 16364140 TI - Task-dependent differences in subjective fatigue scores. AB - The aim of the present study was to evaluate time-on-task effects on subjective fatigue in two different tasks of varying monotony during night-time testing (20:00 to 4:00 hours) in a sleep deprivation intervention. The experiment included eight test runs separated by breaks of approximately 20 min. Twenty healthy volunteers performed a driving simulator and the Mackworth clock vigilance task in four of the test runs each. Sequence of tasks was varied across subjects. Before and after each task, subjective sleepiness was assessed by means of the Karolinska sleepiness scale and subjective fatigue was rated on the Samn Perelli checklist. Fatigue and sleepiness significantly increased over the course of the night. Both tasks led to an increase in fatigue and sleepiness across test runs. However, this time-on-task effect was larger in the vigilance than in the driving simulator task. It is important to note that fatigue and sleepiness in one test run were not influenced by the task performed in the preceding test run, that is there were no cross-over effects. The results suggest that time-on-task effects superimpose circadian and sleep-related factors affecting fatigue. They depend on the monotony of the task and can be quantified by means of a design including separate test runs divided by breaks. PMID- 16364141 TI - Towards a valid, reliable measure of sleep effort. AB - A frequent clinical observation is that patients with insomnia strive to control their sleep. However, sleep is an involuntary physiological process, which cannot be placed under full voluntary control. Therefore, direct, voluntary attempts to control sleep may actually exacerbate and perpetuate insomnia. To date, no reliable scale has been available to test this hypothesis directly. Moreover, while sleep effort is a core International Classification of Sleep Disorders- Revised criterion for psychophysiological insomnia, clinicians lack a reliable measure with which to assess the construct. In this initial scale validation study, we present psychometric data for the Glasgow Sleep Effort Scale based on a relatively small but representative sample of patients with insomnia and good sleepers. The clinical and research value of the new scale is discussed and future research directions are described. PMID- 16364142 TI - Sleep loss-related decrements in planning performance in healthy elderly depend on task difficulty. AB - Prefrontal cortex (PFC)-related functions are particularly sensitive to sleep loss. However, their repeated examination is intricate because of methodological constraints such as practice effects and loss of novelty. We investigated to what extent the circadian timing system and the sleep homeostat influence PFC-related performance in differently difficult versions of a single task. Parallel versions of a planning task combined with a control group investigation were used to control for practice effects. Thirteen healthy volunteers (five women and eight men, range 57-74 years) completed a 40-h sleep deprivation (SD) and a 40-h multiple nap protocol (NAP) under constant routine conditions. Each participant performed 11 easy and 11 difficult task versions under either SD or NAP conditions. The cognitive and motor components of performance could be distinguished and analysed separately. Only by thoroughly controlling for superimposed secondary factors such as practice or sequence effects, could a significant influence of circadian timing and sleep pressure be clearly detected in planning performance in the more difficult, but not easier maze tasks. These results indicate that sleep loss-related decrements in planning performance depend on difficulty level, and that apparently insensitive tasks can turn out to be sensitive to sleep loss and circadian variation. PMID- 16364143 TI - Noradrenergic, serotonergic and GABAergic antagonists injected together into the XII nucleus abolish the REM sleep-like depression of hypoglossal motoneuronal activity. AB - Recently, we reported that the suppression of hypoglossal (XII) motoneuronal activity that occurs during the carbachol-induced, rapid eye movement (REM) sleep like state is abolished by the microinjection into the XII nucleus of a drug mix that antagonizes aminergic excitation and amino acid-mediated inhibition (prazosin, methysergide, bicuculline and strychnine). We now assess the role of glycinergic inhibition in the depression of XII motoneuronal activity and estimate the distribution of the antagonists around the XII nucleus at the time when they are effective. Towards the first goal, REM sleep-like episodes were elicited in urethane-anesthetized rats by 10 nl carbachol microinjections into the dorsomedial pons prior to, and at different times after, combined microinjections into the XII nucleus of only three antagonists (strychnine omitted). As in our previous study, the carbachol-induced depression of XII activity was abolished during tests performed 42-88 min after the antagonists, whereas other characteristic effects of carbachol (appearance of hippocampal theta, cortical activation, decreased respiratory rate) remained intact. The depressant effect of carbachol on XII motoneurons partially recovered after 2.5 h. Towards the second goal, using a drug diffusion model, we determined that the tissue concentrations of the antagonists at the time when they were effective were within the range of their selective actions, and the drugs acted within 0.9 1.4 mm from the injection sites, thus within a space containing XII motoneurons and their dendrites. We conclude that antagonism of alpha-adrenergic, serotonergic, and GABA(A) receptors are sufficient to abolish the REM sleep-like atonia of XII motoneurons. PMID- 16364144 TI - Improved detection of obstructive events in childhood sleep apnoea with the use of the nasal cannula and the differentiated sum signal. AB - The efficacy of the nasobuccal thermistor (NT) was compared with the nasal pressure cannula (NC) and the calibrated, time-differentiated respiratory inductance plethysmography sum signal (DS) in the detection of obstructive events in children during polysomnography (PSG). The overnight PSG of 20 consecutive referrals were selected for analysis. Obstructive events were scored in each study three times by one operator using a blinded procedure whereby either the NT, the NC or the DS was visible. The standard PSG channels were also visible. SPSS software was used for statistical analysis. Twenty patients aged 5 weeks to 16 years were studied. Agreement in obstructive apnoea-hypopnoea index (OAHI) was highest between the NT and NC, and the NC and DS. The NC signal was significantly more likely to be uninterpretable than the NT (P = 0.02) and this did not correlate with age. Event detection by the NT was significantly improved by the addition of either the NC (P = 0.01) or the DS (P = 0.001), while the NC stood alone unless the DS was added (P = 0.02). There was no significant difference in OAHI by the NC versus the DS. The NC detected significantly more OA than the NT or the DS (P = 0.04), while the DS trended towards detecting more OH. There was no significant difference in OAHI between any combination pair. The nasal cannula and differentiated sum signal perform better as measures of paediatric airflow than the NT. To optimize the detection of obstructive events in children we recommend using at least one, if not both these methods in paediatric sleep laboratories. PMID- 16364145 TI - Frequency of narcolepsy symptoms and other sleep disorders in narcoleptic patients and their first-degree relatives. AB - Narcolepsy is a rare neurological sleep disorder affecting around 0.05% of the general population. Genetic factors are known to have an important role in narcolepsy. However, because of its very low prevalence, it is difficult to have groups of comparison between first-degree relatives and general population subjects in order to identify a specific spectrum of disorders in these families. Consequently, from 157 Italian patients with narcolepsy, 263 first-degree relatives were recruited, two refused to participate. These family members were compared with a matched group of 1071 subjects selected from a sample of 3970 subjects representative of the general population of Italy (46 million inhabitants). Finally, 68 spouses of narcoleptic patients were used to assess for possible role of environmental factors. All subjects were interviewed by telephone using the Sleep-EVAL system. Nineteen cases of narcolepsy were discovered among the first-degree relatives of 17 probands (10.8%). Compared with the general population subjects, the relative risk of narcolepsy among female first-degree relatives was of 54.4 and of 105.1 among male first-degree relatives. First-degree relatives were also at higher risk for idiopatic hypersomnia (OR: 23.0), obstructive sleep apnea syndrome (OR: 6.8), adjustment sleep disorder (OR: 4.0), insufficient sleep syndrome (OR: 7.0), circadian rhythm disorders (OR: 2.5), REM behavior disorder (OR: 4.4), and sleep talking (OR: 2.0). The vulnerability to sleep disorders is very high in first-degree relatives and the link with different expressivity and severity of hypersomnia can be confirmed. PMID- 16364146 TI - Variability and predictability in sleep patterns of chronic insomniacs. AB - Sleep of chronic insomniacs is often characterized by extensive night-to-night variability. To date, no study has examined this variability with long series of daily sleep data. The present study examined night-to-night variability with a sample of 106 participants meeting DSM-IV diagnostic criteria for persistent primary insomnia. Participants completed daily sleep diaries for an average of 31 days (range: 18-56). Sleep efficiency, sleep onset latency and wake after sleep onset were derived from this measure. Despite evidence of extensive night variability, results showed that sleep patterns could be classified in three clusters. The first one was characterized by a high probability of having poor sleep, the second one by a low and decreasing probability, and the third one by a constant median probability of having a poor sleep, which is an unpredictable sleep pattern. In the first cluster, poor sleep was expected each night for patients with a predominance mixed insomnia including the three insomnia subtypes. In the second cluster, patients presented moderate insomnia, sleep onset latency below the threshold level and a predominance of sleep-maintenance insomnia. In the third pattern, poor nights seemed unpredictable for patients with moderate to severe insomnia associated with the lowest proportion of sleep maintenance insomnia. Overall, sleep was predictable for about two-thirds of individuals, whereas it was unpredictable for about one-third. These findings confirm the presence of extensive variability in the sleep of chronic insomniacs and that poor sleep may be predictable for some of them. Additional research is needed to characterize those sleep patterns in terms of clinical features and temporal course. PMID- 16364147 TI - Diurnal spectral EEG fluctuations in narcoleptic patients during rest and reaction time tasks. AB - Narcolepsy is associated with lowered vigilance. Diurnal variation in vigilance appears altered, but the exact nature of this change is unclear. It was hypothesized that the homeostatic sleep drive is increased in narcolepsy. Decreased levels of vigilance are reflected in low frequency band power in the electroencephalogram (EEG), so these frequencies were expected to be increased in the narcolepsy group. Furthermore, it was expected that low frequency power should increase over the day. Narcoleptic patients and healthy controls participated (36 participants in total); they were not allowed to take medication or naps on the experimental day. EEG was measured at 9:00, 11:00, 13:00, 15:00, and 17:00 hours, during rest and during reaction time tasks. In the narcolepsy group, alpha power was lower at rest at all times. Delta and theta power during rest and task performance increased steadily over the day in this group, from 11:00 hours onwards. Additionally, in the narcolepsy group beta1 and beta2 power during rest appeared increased at the end of the day. The effects in the lower frequency bands strongly suggest that vigilance is low at all times. The progressive increase in low frequency power indicates that the sleep drive is enhanced. It is not clear whether this pattern reflects an extreme state of low vigilance, or a pathological brain condition. The effects in the higher frequencies suggest that narcoleptic patients may make an effort to counteract their low vigilance level. PMID- 16364148 TI - Agrypnia excitata in a patient with progeroid short stature and pigmented Nevi (Mulvihill-Smith syndrome). AB - We report the video-polysomnographic sleep characteristics of a 25-year-old woman with the Mulvihill-Smith syndrome, a rare clinical condition characterized by progeria-like aspect, peculiar multiple pigmented nevi, low stature, and cognitive impairment. Among the various exams, two overnight video polysomnographic recordings were carried out; moreover, cerebral MRI and molecular analysis of the prion protein gene (PRNP) were also performed. The video-polysomnographic recordings showed the absence of clear sleep episodes but the presence of periods during which the patient had poor contact with the environment, stereotyped afinalistic movements of the upper limbs and hands, irregular or periodic breathing (with central apnea episodes), heart rate arrhythmia, and rapid eye movements. Cerebral MRI showed only diffuse mild enlargement of the cortical sulci and the molecular genetics analysis of the PRNP was normal. Our clinical and neurophysiological study seems to indicate that a particular condition of severe sleep disruption, similar to some extent to that reported in the fatal familial insomnia and in the Morvan fibrillary chorea, which has been indicated as Agrypnia Excitata in recent literature, might be associated with the Mulvihill-Smith syndrome. The inclusion of a detailed study on the sleep characteristics of eventual additional patients will certainly help our understanding of this rare condition. PMID- 16364151 TI - Regulatory T cells: which role in the pathogenesis and treatment of allergic disorders? AB - Several recent data suggest the importance of different types of cells in the regulation of T-cell effector-mediated immune responses. However, a unique specific marker for these cells has not yet been identified. Moreover, in addition to a dedicated functional lineage, even a 'plastic' phenotype of regulatory T cells seems to exist. The lack of a unique specific marker for regulatory T cells, as well as their heterogeneity, make it difficult to determine whether a defect of regulatory T cells plays a role in the pathogenesis of common allergic disorders. Novel therapeutic strategies based on the induction or potentiation of regulatory T cells able to hamper allergic inflammatory reactions are desirable, but their possible efficacy and safety are not yet known. At present, therapeutic strategies able to induce an immune deviation of allergen-specific type 2 T-helper responses to a less polarized phenotype rather than a general immune suppression appear more realistic. PMID- 16364150 TI - Themes in allergy. PMID- 16364153 TI - CD4+ CD30+ T cells perpetuate IL-5 production in Dermatophagoides pteronyssinus allergic patients. AB - BACKGROUND: Airway allergic diseases are regulated by interleukin (IL)-5, which causes infiltration of eosinophils into the bronchial epithelium, and by IL-4 which increases serum immunoglobulin E (IgE) production and promotes CD30 expression on Th cells. CD30 generates a costimulatory signal involved in apoptosis or cell proliferation, depending on the microenvironment. Our aims were: (i) to analyze if CD4+ CD30+ T cells from allergic patients proliferate in response to Dermatophagoides pteronyssinus, and (ii) if upon stimulation this cell population produces IL-4 and IL-5. METHODS: Peripheral blood mononuclear cell (PBMC) from 17 allergic rhinitis and mild allergic asthma patients and 12 healthy nonallergic individuals were stimulated with allergen in the presence or absence of anti-IL-4, anti-IL-5 or anti-IL-4Ralpha monoclonal antibodies (mAbs). TdT-mediated dUTP nick end-labeling (TUNEL) assay, 7-aminoactinomycin-D (7-AAD) intercalation, and flow cytometry were used to determine the CD4+ CD30+ blasts percentage, cell proliferation, apoptosis, and intracellular cytokines after 7 culture days. RESULTS: Cell proliferation induced with allergen showed that 90% of the allergen-stimulated blasts were CD4+, 50% of which were CD30+. Allergen stimulated PBMC showed a progressive increase (mean: from 7% to 23%) of CD4+ CD30+IFN-gamma+ and CD4+ CD30+IL-4+ blasts which diminished (mean: 6%) after 5 culture days. In contrast, CD4+ CD30+IL-5+ blasts showed a continuous progression (from 12% to 24%) that maintained after 7 culture days. The vast majority of CD4+ CD30+ blasts were negative to 7-AAD or TUNEL. Additionally, a significant decrease (34%) was observed in the number of CD4+ CD30+ blasts when IL-4 was neutralized. CONCLUSIONS: These data suggest that specific allergen stimulation of PBMC isolated from allergic patients generates a nonapoptotic CD4+ CD30+ blast subset that produces IL-5. PMID- 16364152 TI - CD28/CTLA-4--CD80/CD86 and ICOS--B7RP-1 costimulatory pathway in bronchial asthma. AB - Costimulatory molecules are cell surface glycoproteins that can direct, modulate and fine-tune T-cell receptor signals. The B7-1/B7-2--CD28/CTLA-4 and ICOS-B7RP-1 pathway provides key second signals that can regulate the activation, inhibition and fine-tuning of T-lymphocyte responses. The expression of B7-1/B7-2--CD28/CTLA 4 molecules on clinical samples from patients with asthma have been well studied, and the results indicate that different extents of these molecules are expressed on the surface of various cells, and that the concentrations of soluble form of these molecules are elevated in the sera of patients with asthma. There is a burst of papers describing an important role for B7-1/B7-2--CD28/CTLA-4 pathway in the Th1/Th2 balance. Similarly, ICOS stimulates both Th1 and Th2 cytokine production but may have a preferential role in Th2 cell development. Moreover, The B7-1/B7-2-CD28/CTLA-4 and ICOS-B7RP-1 pathway has been suggested of being involved in the development of airway inflammation and airway hyperresponsiveness. Further study of the functions of the pathways within the CD28/CTLA-4--CD80/CD86 and ICOS--B7RP-1 superfamily individually and their interplay should provide insights into the pathogenesis of asthma, and has great therapeutic potential for treatment of asthma. PMID- 16364154 TI - Variation of T-cell activation in allergic subjects during natural pollen exposure. AB - BACKGROUND: Allergic inflammation is characterized by a Th2 activation. However, little is known about dynamics of T-cell cytokine production during natural allergen exposure. The aim of this study was to assess the Th1/Th2 balance in cypress allergic patients compared with controls, and variations of this balance over the pollen season. METHODS: Twenty cypress allergic patients and 10 controls were studied, distributed during two consecutive pollen seasons. Cytokine production was assessed by flow cytometry and ELISA. The variation of cytokine production during the pollen season was analyzed among patients in four occasions, and the preseason values were compared with controls. IL-13 and IFN gamma-containing T cells were assessed among whole blood cells and PBMC. In addition the effect of specific stimulation by Juniperus ashei pollen extract was studied. RESULTS: Compared with controls, IL-13-producing T cells were increased in allergics in any case. By contrast, compared with controls, allergic IFN-gamma producing T cells were decreased in whole blood, but not in PBMC, and were increased after specific stimulation. During the season, an increase in IFN-gamma and a decrease in IL-13-producing T cells occurred in patients, whatever the culture conditions. CONCLUSION: These results show that the allergic T-cell activation is not limited to a Th2 profile: allergen-stimulated T cells are able to produce IFN-gamma at baseline, and the Th1/Th2 ratio increases during the pollen season. PMID- 16364155 TI - T cell and eosinophil activation in mild and moderate atopic and nonatopic children's asthma in remission. AB - BACKGROUND: Inflammation in the pathogenesis of asthma is associated with products of activated T cells and eosinophils. The aim of this study was to determine whether ongoing inflammation persists in children with different phenotypes of asthma despite the disease in remission. METHODS: Serum samples were collected from 68 children with atopic or nonatopic asthma in remission and from 15 healthy children. Soluble interleukin-2 receptor (sIL-2R), IL-2 and IL-4 were examined by using an enzyme-linked immunosorbent assay. Total and specific immunoglobulin E, and eosinophil cationic protein (ECP) were analysed by fluoroimmunoassay (Pharmacia CAP System). RESULTS: In patients with moderate persistent atopic asthma, sIL-2R was increased significantly when compared with mild persistent atopic asthma (P < 0.05). No changes of sIL-2R were seen in nonatopic asthmatics compared with atopics and controls. The level of IL-2 was elevated in moderate persistent atopic and nonatopic asthmatic children compared with controls (P < 0.05 and P < 0.05 respectively) and compared with mild persistent atopic asthmatics and mild persistent nonatopic asthmatics (P < 0.05 in both cases). The levels of IL-4 in most patients and controls remained below the sensitivity of the assay. Eosinophil cationic protein levels in moderate persistent atopic and nonatopic asthmatics were significantly higher than in mild persistent asthma severity cases (P < 0.001 and P < 0.01 respectively) and in healthy children (P < 0.01 in both cases). CONCLUSION: Changes in the concentration of sIL-2R, IL-2 and ECP reflect increased T cell and eosinophil activity in relation to the level of severity of asthma in atopic and nonatopic children, thereby proving the presence of persistent inflammation despite the absence of disease symptoms. PMID- 16364156 TI - Pholcodine stimulates a dramatic increase of IgE in IgE-sensitized individuals. A pilot study. AB - BACKGROUND: A previous study showed a relation between pholcodine (PHO) consumption, prevalence of IgE-sensitization to PHO, morphine (MOR) and suxamethonium (SUX) and anaphylaxis to neuromuscular blocking agents (NMBA). The purpose of this pilot study was to explore the effect on IgE production, in IgE sensitized and nonsensitized individuals, of exposure to cough syrup and environmental chemicals containing PHO, MOR and SUX related allergenic structures. METHODS: Serum concentrations of IgE and IgE antibodies to PHO, MOR and SUX allergens measured by ImmunoCAP (Pharmacia Diagnostics, Uppsala, Sweden) were followed after intake of cough syrup, or exposure to confectionary and other household chemicals containing various amounts of substances cross-reacting with PHO, MOR and SUX. RESULTS: Cough syrup containing PHO gave, in sensitized individuals, within 1-2 weeks, an increase of IgE of 60-105 times and of IgE antibodies to PHO, MOR and SUX in the order of 30-80 times. The tested confectionary did not have any similar stimulating effect but seemed to counteract the expected decrease of IgE. No effect was seen in nonsensitized individuals. The PHO stimulated IgE showed a nonspecific binding to ImmunoCAP with common allergens and glycine background ImmunoCAP that was up to 10-fold higher than that of monomeric myeloma-IgE at twice the concentration. CONCLUSIONS: It seems as cough syrups containing PHO have a most remarkable IgE boostering effect in persons IgE-sensitized to PHO, MOR and SUX related allergens. Household chemicals containing such allergenic epitopes seem capable of some, minor, stimulation. PMID- 16364157 TI - Cytokine and Ig-production by CG-containing sequences with phosphorodiester backbone and dumbbell-shape. AB - BACKGROUND: Immunostimulatory oligodeoxynucleotides (CpG-ODN) usually contain phosphorothioate (PS) backbones for nucleotide protection, which may result in some nonspecific side-effects like prolongation of coagulation time. OBJECTIVE: The aim of the present study was to investigate the immunomodulatory potential of DNA molecules without PS backbones. Thus, we designed phosphorodiester (PO) molecules with a dumbbell-like covalently-closed structure (dSLIM-30L1). METHODS: We analyzed their effects on peripheral blood mononuclear cells (PBMC) from spontaneous high and low immunoglobulin (Ig)E producer (allergic and nonallergic donors) in comparison with linear CpG-ODN (lin-30L1) with PS backbones, using enzyme-linked immunosorbent assay and flow cytometry. RESULTS: We observed a decrease of spontaneous IgE levels in PBMC from high IgE producer of approximately 27% with both dSLIM-30L1 and lin-30L1. In addition, both molecules enhanced the production of IgA, IgM and IgG1/IgG2, but with a slightly different pattern. Both molecules stimulated the secretion of the T(H)1-like cytokines interleukin (IL)-2, interferon-gamma and IL-12p40 and the pro-inflammatory cytokine IL-6. The immunomodulatory potential of dSLIM-30L1 and lin-30L1 was also effective in PBMC from nonallergic donors, as was confirmed for IL-2, IL-12p40, IgG1/IgG2 and IgM. CONCLUSION: Our data show an inhibition of IgE production but also enhancement of the inflammatory cytokine response in PBMC from allergic and nonallergic donors by covalently-closed PO-based dSLIM-30L1 with a pattern similar to that of linear PS-based lin-30L1, while avoiding PS-modifications and thus PS-mediated side-effects. Whether such molecules are useful for the treatment of allergic diseases will need further clarification by appropriate in vivo studies. PMID- 16364158 TI - Role of TLR4 in allergic sensitization to food proteins in mice. AB - Allergic sensitization to food proteins and other allergens is increasing in prevalence. One hypothesis for this increase is that the decreased rate of infections or exposure to microbial products leaves the immune system susceptible to inappropriate reactivity to innocuous antigens through the lack of development of regulatory cells. We hypothesized that constitutive Toll-like receptor (TLR)4 signaling (presumably via the commensal flora) could inhibit the development of allergic sensitization to food proteins. We tested this hypothesis by sensitizing TLR4+ and TLR4- mice on two genetic backgrounds, C3H and BALB/c, to two common food allergens [beta-lactoglobulin (betaLG) and peanut (PN)]. B-cell responses were not significantly influenced by TLR4 status. T-cell responses were Th2 skewed in TLR4-deficient C3H mice compared with TLR4 sufficient C3H mice, but this pattern of Th2 skewing was not observed in TLR4-deficient mice on a BALB/c background. In anaphylaxis-susceptible C3H mice, TLR4 deficiency was associated with increased severity of anaphylaxis to PN, and decreased severity of anaphylaxis to betaLG. In anaphylaxis-resistant BALB/c mice, TLR4 deficiency was not sufficient to render mice susceptible to PN-induced anaphylaxis. We conclude that although TLR4 status can influence T-cell responses and anaphylaxis severity, the nature of the influence is highly antigen- and strain-dependent. PMID- 16364159 TI - Comparison of roflumilast, an oral anti-inflammatory, with beclomethasone dipropionate in the treatment of persistent asthma. AB - BACKGROUND: Roflumilast is an oral, once-daily phosphodiesterase 4 inhibitor with anti-inflammatory activity in development for the treatment of asthma. Roflumilast was compared with inhaled beclomethasone dipropionate (BDP) in patients with asthma. METHODS: In a double blind, double-dummy, randomized, noninferiority study, 499 patients (forced expiratory volume in 1 s [FEV1] = 50 85% predicted) received roflumilast 500 microg once daily or BDP 200 microg twice daily (400 microg/day) for 12 weeks. Lung function and adverse events were monitored. RESULTS: Roflumilast and BDP significantly improved FEV1 by 12% (270 +/- 30 ml) and 14% (320 +/- 30 ml), respectively (P < 0.0001 vs baseline). Roflumilast and BDP also significantly improved forced vital capacity (FVC) (P < 0.0001 vs baseline). There were no significant differences between roflumilast and BDP with regard to improvement in FEV1 and FVC. Roflumilast and BDP showed small improvements in median asthma symptom scores (-0.82 and -1.00, respectively) and reduced rescue medication use (-1.00 and -1.15 median puffs/day, respectively; P < 0.0001 vs baseline). These small differences between roflumilast and BDP were not considered clinically relevant. Both agents were well tolerated. CONCLUSIONS: Once daily, oral roflumilast 500 microg was comparable with inhaled twice-daily BDP (400 microg/day) in improving pulmonary function and asthma symptoms, and reducing rescue medication use in patients with asthma. PMID- 16364160 TI - Are overweight asthmatics more difficult to control? AB - The relationship between asthma and obesity appears to be quite complex. The aim of this study was to assess the effect of excess weight on asthma control evolution in a cohort of asthmatics. A prospective database was set up, which enrolled adult asthmatics with persistent (mild, moderate or severe) asthma. The control of asthma was defined as a binary variable, acceptable or unacceptable. In order to evaluate the effect of body mass index (BMI; <25 or > or =25), data were analysed using a continuous time homogeneous Markov model in which the forces ruling the transition between the two health states were estimated. The following confounding covariates were also evaluated in the model: severity of asthma, current treatment with oral corticosteroids (OCS) and history of OCS over the year preceding inclusion. About 406 asthmatics were included who made a total of 1639 consultations; the median length of follow up was 182 days. Using a univariate model, overweight patients had a lower risk of transiting from the unacceptable to the acceptable health state (RR = 0.45; P < 0.01). The effect of weight remained significant (RR = 0.53; P < 0.01) in the multivariate model including the other covariates. Moreover, transition probabilities stabilized more rapidly for patients with BMI < 25 (200 vs 300 days). In this study, we thus demonstrated that there is an association between excess weight and transition from unacceptable to acceptable control. Because control of asthma clearly drives asthma management, this finding has consequences for defining original new strategies for managing asthma in overweight patients. PMID- 16364161 TI - Nitrogen washout slope in poorly controlled asthma. AB - Short-term control of asthma is often lacking even though forced expiratory volume in 1 s (FEV1) remains above normal value. Small airways are a potential key site of persistent inflammation and structural modification. Noninvasive assessment of small airways was found to be difficult, but the computerized single breath nitrogen washout test (SBNT) has been recently successfully reintroduced with this aim. Twenty-four asthmatics (13 females) of various severity but with normal FEV1 were compared with 24 healthy volunteers (13 females) and studied at steady-state after bronchodilatation (400 microg salbutamol). Spirometric values, plethysmographic data, phase III (slope of phase III of the SBNT, dN2) and IV [closing volume (CV), with closing capacity (CC) = CV + residual volume (RV)] of the SBNT were checked. Asthma severity, recent control, exacerbation rate, and therapy requirements were assessed on the basis of validated questionnaires (ACQ) and international guidelines. Patients were prospectively pooled into two equal groups according to their exacerbation rate. The reproducibility of the measurements obtained on 2 following days was assessed. All plethysmographic values, except total lung capacity (TLC), differentiated asthmatic patients from controls. The CC/TLC [124 (117-148) vs 117 (112-123), P = 0.04] and dN2 [110 (99-190) vs 94 (75-111), P = 0.02] were increased in asthma. The dN2 was significantly increased in patients with frequent exacerbations [100 (83-105) vs 195 (141-212), P = 0.0005]. A correlation was obtained between dN2 and recent asthma control (rho: 0.62; P = 0.003), number of exacerbations (rho: 0.71, P = 0.0008), and RV/TLC (rho: 0.49, P = 0.026). This study demonstrated that ventilation inequalities assessed by dN2 represent an important indicator of poor asthma control and high exacerbation rate in high symptom perceivers. New therapies focused on small airways should now be developed. PMID- 16364163 TI - ALOX5 promoter genotype, asthma severity and LTC production by eosinophils. AB - BACKGROUND: The number of Sp1-Egr1 binding tandem repeats at the ALOX5 promoter influences gene transcription and may modify the response to anti-leukotriene treatment. The relationship of ALOX5 variants to asthma severity and leukotriene production by eosinophils is unknown. OBJECTIVE: To characterize ALOX5 mRNA expression and cysteinyl-leukotriene production by eosinophils from individuals bearing ALOX5 promoter deletional variants and their association with the severity of childhood asthma. METHODS: Eosinophils from adult asthmatics bearing only variant alleles (with other than five tandem repeats on both chromosomes, non5/non5) or no variant alleles (5/5) were cultured in vitro and ALOX5 expression and leukotriene secretion were measured. A total of 621 children with mild or moderate-severe asthma were genotyped at the ALOX5 core promoter. RESULTS: Asthmatics with non5/non5 genotype expressed less ALOX5 mRNA and produced less LTC4 into culture supernatants than 5/5 individuals (6.4 +/- 2.0 and 20.0 +/- 5.0 pg/ml, n = 5; P < 0.05). More asthmatic children bearing non5/non5 genotype had moderate-severe asthma than children with the 5/5 genotype (5.3% vs. 1.4%, P = 0.008). Multivariate logistic regression identified ALOX5 promoter genotype as a significant predictor of disease severity (OR = 3.647, 95% CI: 1.146-11.608, P = 0.03). Consistent with these findings, children bearing the non5/non5 genotype had greater bronchomotor response to exercise as measured by the maximum fall after exercise and the area under the exercise curve (P < 0.05 for both). CONCLUSION: Our results suggest that children who express the asthma phenotype despite having a genetic variant that impairs their ability to express ALOX5 have more severe disease and thus are more likely to have asthma symptoms. PMID- 16364162 TI - 3-Nitrotyrosine, a marker of nitrosative stress, is increased in breath condensate of allergic asthmatic children. AB - BACKGROUND: Asthmatic patients have high exhaled nitric oxide (NO) levels. NO mediated inflammatory actions are mainly due to NO conversion into reactive nitrogen species, which can lead to nitrotyrosine formation. The aim of this study was to assess 3-nitrotyrosine (3-NT) levels in exhaled breath condensate (EBC) of asthmatic and healthy children and to investigate whether there is any relationship with exhaled NO (FE(NO)) and lung function. METHODS: The study included 20 asthmatic children (10 steroid-naive with intermittent asthma, 10 steroid-treated with unstable persistent asthma) and 18 healthy controls. They underwent FE(NO) measurement, EBC collection and spirometry. 3-NT was measured by a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method in isotopic dilution. RESULTS: The median EBC concentration of 3-NT (expressed as nitrotyrosine/tyrosine ratio x 100) in asthmatic children was fivefold higher than in healthy subjects [0.23% (0.12-0.32) vs 0.04% (0.02-0.06), P < 0.001] with no difference between steroid-naive and unstable steroid-treated asthmatic patients. FE(NO) levels were higher in asthmatic [44.6 ppb (36.0-66.0)] than in healthy children [7.5 ppb (6.0-8.8), P < 0.001]. No correlation was found among 3 NT, FE(NO) and lung function parameters. CONCLUSION: Nitrotyrosine is high in EBC of asthmatic children and could be considered as a noninvasive marker of nitrosative events in the airways. PMID- 16364164 TI - Sensitization to blackgram in patients with bronchial asthma and rhinitis: clinical evaluation and characterization of allergens. AB - BACKGROUND: Legumes are important causative agents of type I hypersensitivity in south Asia and Europe but such studies are lacking in Indian population. The present study investigates blackgram sensitization in asthma and rhinitis patients and identifies immunoglobulin E (IgE)-binding proteins. METHODS: Respiratory allergy patients were evaluated using standard questionnaire and skin prick tests (SPT) with common foods and aeroallergens. Blackgram-specific IgE level was estimated by enzyme-linked immunosorbent assay (ELISA) and sensitization was established by a double-blind, placebo-controlled food challenge (DBPCFC). The cross-reactivity of blackgram with other legumes was studied by immunobiochemical methods. RESULT: Of 816 patients, 35 gave history of blackgram hypersensitivity. From these, 16 patients were SPT positive and 14 showed elevated specific IgE (three times of negative control) to blackgram. DBPCFC established blackgram sensitivity in four of 14 patients. Immunoblotting with individual patient's sera recognized eight most prevalent allergens of 78, 56, 47, 43, 40, 30, 28 and 16 kDa. Roasted blackgram showed six major allergens whereas 47, 43 and 28 kDa proteins retained IgE reactivity upon boiling. Blackgram extract required 14 ng of self protein for 50% ELISA inhibition whereas roasted and boiled blackgram required 16 and 120 ng protein. ELISA and immunoblot inhibition show partial inhibition to blackgram proteins by lentil, limabean and pea. CONCLUSION: Blackgram induces IgE-mediated reactions in 1.7% of asthma and rhinitis patients and contains eight major IgE-binding components, of which six retained IgE reactivity after roasting. Blackgram shares allergenicity with lentil and limabean. PMID- 16364165 TI - Changes in airway inflammation following nasal allergic challenge in patients with seasonal rhinitis. AB - BACKGROUND: Seasonal allergic rhinitis could predispose to the development of chronic bronchial inflammation as observed in asthma. However, direct links between nasal inflammation, bronchial inflammation and airway responsiveness in patients with seasonal allergic rhinitis and without asthma are not fully understood. The aim of this study was to analyse the changes induced by allergic nasal challenge outside the pollen season in airway responsiveness and bronchial inflammation of patients with seasonal allergic rhinitis. METHODS: Nine patients were evaluated after either grass pollens or placebo nasal challenge in a randomized cross-over double-blinded trial. Nasal parameters were recorded hourly and airway responsiveness was assessed by methacholine challenge. Cytological examinations and cytokine measurements were performed in nasal lavage and induced sputum. Eosinophil activation was investigated by eosinophil-cationic protein expression and secretion. RESULTS: Airway responsiveness was increased after allergic nasal challenge. Total eosinophils and eosinophils expressing eosinophil cationic protein were increased in induced sputum after allergic nasal challenge. Both eosinophil number and eosinophil-cationic protein concentration in induced sputum were correlated to methacholine responsiveness. CONCLUSIONS: These results suggest that eosinophils participate to the bronchial inflammation in patients with seasonal allergic rhinitis following allergic nasal challenge outside the pollen season and might explain changes in airway responsiveness. PMID- 16364166 TI - High-efficiency vacuum cleaners increase personal mite allergen exposure, but only slightly. AB - BACKGROUND: High-efficiency particulate-arrest-filter vacuum cleaners are recommended to allergy sufferers although their use increases personal cat allergen exposure. We aimed to measure personal mite allergen exposure during vacuum cleaning by nasal air sampling and to compare exposures while vacuuming and emptying the vacuum cleaner bag. METHODS: Five brand new high-efficiency vacuum cleaners were selected. An old, previously used vacuum cleaner with its original microfilter in situ was used as a control. Nasal air samples were taken prior to and during vacuum cleaning in 10 homes. Samples were processed by HALOgen assay. Personal mite and cat allergen exposure was measured as the dust compartments were emptied. RESULTS: There was an increase in personal mite allergen exposure with vacuum cleaning, which approached significance (P = 0.058). There was no difference between the high-efficiency vacuum cleaners and the control vacuum cleaner (P = 0.141). When the dust compartments were emptied, personal mite and cat allergen exposure increased (P < 0.02). CONCLUSIONS. The increase in personal mite allergen exposure while vacuum cleaning in an area with mild to moderate mite allergen exposure is small. High-efficiency vacuum cleaners confer no benefit and cannot currently be recommended to allergy sufferers as a means of reducing personal mite allergen exposure. PMID- 16364167 TI - Turnip rape and oilseed rape are new potential food allergens in children with atopic dermatitis. AB - BACKGROUND: When skin prick testing (SPT) young children with atopic dermatitis (AD) for suspected food allergy, we frequently found positive reactions with turnip rape (Brassica rapa) and oilseed rape (Brassica napus). We performed food challenge to examine whether these children react clinically to turnip rape. METHODS: A total of 1887 children were screened with SPTs for sensitization to turnip rape and oilseed rape. Twenty-eight children with clearly positive SPT (> or =5 mm) were first subjected to labial challenge with turnip rape seeds followed, if negative, by open oral challenge for up to 7 days. Twenty-five children with AD but negative SPT to turnip rape and oilseed rape served as controls. RESULTS: Two-hundred and six (10.9%) children had positive SPT to turnip rape and/or oilseed rape. Twenty-five (89%) of 28 children showed a positive challenge reaction to turnip rape. Seventeen reacted with labial whealing, and eight in oral challenge with facial urticaria, flare-up of AD or abdominal symptoms. All 25 control children remained negative in the labial challenge. CONCLUSIONS: Turnip rape and oilseed rape seem to be new important food allergens in young children with AD. The modes of exposure to these allergens and the possible routes of sensitization remain to be established. PMID- 16364168 TI - Screening the allergenic repertoires of wheat and maize with sera from double blind, placebo-controlled food challenge positive patients. AB - BACKGROUND: Food allergy to wheat and maize is an increasing factor of deterioration of life quality, especially childhood and can, in rare cases, even induce anaphylaxis. Although omega-5 gliadin from wheat and maize lipid transfer protein have been characterized as major cereal allergens on the molecular level, the list of food allergens is far to be complete. METHODS: To identify the IgE binding repertoires of wheat and maize we screened respective cDNA libraries displayed on phage surface with sera from patients with a confirmed food allergy. The study included six patients with a positive double-blind, placebo-controlled food challenge (DBPCFC) to wheat, nine patients with a positive DBPCFC to maize, and six patients with anaphylactic reactions after ingestion of wheat. RESULTS: The enriched sequences encoding IgE-binding proteins showed heterogeneous repertoires for both, wheat and maize. The selected wheat repertoire yielded 12, the maize repertoire 11 open reading frames. Among these we identified allergens belonging to already characterized allergens families, such as gliadin, profilin and beta-expansin. Besides, we found novel proteins with high cross-reactive potential, such as thioredoxins, as well as sequences that had so far not been related to cereal allergy at all. The IgE-binding capacity of some selected proteins was evaluated in vitro and cross-reactivity was demonstrated by competition ELISA. CONCLUSION: With regard to the heterogeneity of the characterized sequences as well as to the biochemical nature of the new allergens detected we conclude that wheat and maize-related food allergy is more complex than so far anticipated. PMID- 16364169 TI - Cysteinyl-leukotrienes contribute to sputum eosinophil chemotactic activity in asthmatics. AB - BACKGROUND: Cysteinyl-leukotrienes are lipid derived mediators involved in asthma. They are able to stimulate eosinophil chemotaxis in vitro. Induced sputum from asthmatics has been shown to contain eosinophil chemotactic activity. The purpose of our study was to evaluate the contribution of cysteinyl-leukotrienes to sputum eosinophil chemotactic activity in asthmatics and to seek whether there might be differences between asthmatics free of inhaled corticosteroids vs those regularly receiving this treatment. METHODS: Twenty-two patients (11 corticosteroid free, mean FEV1 99% predicted, 11 corticosteroid-treated, mean FEV1 77% predicted) recruited from our asthma clinic underwent a sputum induction. Sputum was processed according to standard procedure. Eosinophil chemotactic activity contained in the fluid phase was assessed using Boyden microchamber model and expressed as chemotaxis index (CI). Cysteinyl-leukotrienes were measured in sputum supernatant by ELISA and their role in sputum eosionophil chemotactic activity was evaluated by using montelukast, a selective antagonist of a cys-LT1 receptor. RESULTS: Cysteinyl-leukotrienes were well detectable in sputum supernatants from both steroid-naive (247 +/- 42 pg/ml) and steroid treated (228 +/- 26 pg/ml) asthmatics. Sputum eosinophil chemotactic activity was indiscriminately present in both corticosteroid-naive (CI: 2.61 +/- 0.22) and corticosteroid-treated (2.98 +/- 0.35) asthmatics. Montelukast (100 microM) significantly inhibited the eosinophil chemotactic activity in both groups achieving a mean inhibition of 54.2 +/- 9.2% (P < 0.001) and 64.7 +/- 7.8% (P < 0.001) in steroid-naive and steroid-treated asthmatics respectively. CONCLUSION: Cysteinyl-leukotrienes actively participate in sputum eosinophil chemotactic activity found in asthmatics irrespective of whether they are or not under treatment with inhaled corticoids. PMID- 16364171 TI - Sensitization to pig albumin and gamma-globulin responsible for occupational respiratory allergy. PMID- 16364170 TI - Fungal contamination of bedding. AB - BACKGROUND: It is currently believed that most fungal exposure occurs external to the home. AIMS: To enumerate the fungal flora of used synthetic and feather pillows and the dust vacuumed from them, in the UK. METHODS: 10 pillows aged between 1.5 and >20 years in regular use were collected and quantitatively cultured for fungi. Swatches were taken from nine sections of the pillow and dust was also collected by vacuum from five pillows. Pillow vacuuming was carried out prior to pillow culture. All were cultured at room temperature, 30 and 37 degrees C for 7 days in broth before plating, and a subset were also cultured for 24 h in broth and then plated. Fungi were identified by standard morphological methods. RESULTS: The commonest three species isolated were Aspergillus fumigatus (n = 10), Aureobasidium pullulans (n = 6) and Rhodotorula mucilaginosa (n = 6). Another 47 species were isolated from pillows and vacuum dust. The number of species isolated per pillow varied from 4 to 16, with a higher number from synthetic pillows. Compared with the nonallergenic A. pullulans, more A. fumigatus was found in synthetic than feather pillows. CONCLUSIONS: We have examined pillows for fungal contamination, and show that the typical used pillow contains a substantial load of many species of fungi, particularly A. fumigatus. Given the time spent sleeping, and the proximity of the pillow to the airway, synthetic and feather pillows could be the primary source of fungi and fungal products. This has important implications for patients with respiratory disease, and especially asthma and sinusitis. PMID- 16364172 TI - Multiple drug intolerance including etoricoxib. PMID- 16364173 TI - Vesicular and bullous eczema in response to intravenous immunoglobulins (IVIG). PMID- 16364174 TI - Bronchospasm induced by inhalant corticosteroids: the role of ethanol. PMID- 16364175 TI - Internet-based Profiler system as integrative framework to support translational research. AB - BACKGROUND: Translational research requires taking basic science observations and developing them into clinically useful tests and therapeutics. We have developed a process to develop molecular biomarkers for diagnosis and prognosis by integrating tissue microarray (TMA) technology and an internet-database tool, Profiler. TMA technology allows investigators to study hundreds of patient samples on a single glass slide resulting in the conservation of tissue and the reduction in inter-experimental variability. The Profiler system allows investigator to reliably track, store, and evaluate TMA experiments. Here within we describe the process that has evolved through an empirical basis over the past 5 years at two academic institutions. RESULTS: The generic design of this system makes it compatible with multiple organ system (e.g., prostate, breast, lung, renal, and hematopoietic system,). Studies and folders are restricted to authorized users as required. Over the past 5 years, investigators at 2 academic institutions have scanned 656 TMA experiments and collected 63,311 digital images of these tissue samples. 68 pathologists from 12 major user groups have accessed the system. Two groups directly link clinical data from over 500 patients for immediate access and the remaining groups choose to maintain clinical and pathology data on separate systems. Profiler currently has 170 K data points such as staining intensity, tumor grade, and nuclear size. Due to the relational database structure, analysis can be easily performed on single or multiple TMA experimental results. The TMA module of Profiler can maintain images acquired from multiple systems. CONCLUSION: We have developed a robust process to develop molecular biomarkers using TMA technology and an internet-based database system to track all steps of this process. This system is extendable to other types of molecular data as separate modules and is freely available to academic institutions for licensing. PMID- 16364176 TI - Difficulties associated with outpatient management of drug abusers by general practitioners. A cross-sectional survey of general practitioners with and without methadone patients in Switzerland. AB - BACKGROUND: In Switzerland, general practitioners (GPs) manage most of the patients receiving methadone maintenance treatment (MMT). METHODS: Using a cross sectional postal survey of GPs who treat MMT patients and GPs who do not, we studied the difficulties encountered in the out-patient management of drug addicted patients. We sent a questionnaire to every GP with MMT patients (556) in the French-speaking part of Switzerland (1,757,000 inhabitants). We sent another shorter questionnaire to primary care physicians without MMT patients living in the Swiss Canton of Vaud. RESULTS: The response rate was 63.3%. The highest methadone dose given by GPs to MMT patients averaged 120.4 mg/day. When asked about help they would like to be given, GPs with MMT patients primarily mentioned the importance of receiving adequate fees for the care they provide. Secondly, they mentioned the importance of better training, better knowledge of psychiatric pathologies, and discussion groups on practical cases. GPs without MMT patients refuse to treat these patients mostly for emotional and relational reasons. CONCLUSION: GPs encounter financial, relational and emotional difficulties with MMT patients. They desire better fees for services and better training. PMID- 16364177 TI - Balance of IL-10 and interferon-gamma plasma levels in human visceral leishmaniasis: implications in the pathogenesis. AB - BACKGROUND: Leishmaniasis remains a serious public health problem in several parts of the developing world. Effective prophylactic measurements are hampered by imprecise comprehension of different aspects of the disease, including its immunoregulation. A better comprehension of immunoregulation in human VL may be useful both for designing and evaluating immunoprophylaxis. METHODS: To explore immunoregulatory mechanisms, 20 visceral leishmaniasis (VL) patients were evaluated during active disease and at different periods up to one year after treatment determining their plasma cytokine levels, clinical parameters (palpable spleen and liver) and antibody levels. RESULTS: Elevated plasma levels of IFN gamma and of IL-12 p40 were observed during active disease, significantly decreasing after treatment whereas in vitro Leishmania antigen-stimulated IFN gamma production by PBMC exhibited an inverse pattern being low during disease and increasing steadily thereafter. Absence of IFN-gamma activity is a hallmark of VL. The main candidate for blunting IFN-gamma activity is IL-10, a cytokine highly elevated in plasma with sharp decrease after treatment. Activity of IL-10 is inferred by high levels of anti-Leishmania specific IgG1 and IgG3. TGF-beta had elevated total, but not of active, levels lessening the likelihood of being the IFN-gamma counterpart. Spleen or liver size presented a steady decrease but return to normal values at only 120 days after treatment. Anti-Leishmania IgG (total and subclasses) levels and DTH or Leishmania-stimulated lymphocyte proliferation conversion to positive also present a slow decrease after treatment. IL-6 plasma levels were elevated in only a few patients. CONCLUSION: Taken together our results suggest that IFN-gamma and IL-10 are the molecules most likely involved in determining fate of disease. After treatment, there is a long delay before the immune profile returns to normal what precludes using plasma cytokine levels as criteria of cure as simpler clinical evaluations, as a palpable spleen or liver, can be used. PMID- 16364178 TI - Safer injections following a new national medicine policy in the public sector, Burkina Faso 1995-2000. AB - BACKGROUND: The common failure of health systems to ensure adequate and sufficient supplies of injection devices may have a negative impact on injection safety. We conducted an assessment in April 2001 to determine to which extent an increase in safe injection practices between 1995 and 2000 was related to the increased access to injection devices because of a new essential medicine policy in Burkina Faso. METHODS: We reviewed outcomes of the new medicine policy implemented in 1995. In April 2001, a retrospective programme review assessed the situation between 1995 and 2000. We visited 52 health care facilities where injections had been observed during a 2000 injection safety assessment and their adjacent operational public pharmaceutical depots. Data collection included structured observations of available injection devices and an estimation of the proportion of prescriptions including at least one injection. We interviewed wholesaler managers at national and regional levels on supply of injection devices to public health facilities. RESULTS: Fifty of 52 (96%) health care facilities were equipped with a pharmaceutical depot selling syringes and needles, 37 (74%) of which had been established between 1995 and 2000. Of 50 pharmaceutical depots, 96% had single-use 5 ml syringes available. At all facilities, patients were buying syringes and needles out of the depot for their injections prescribed at the dispensary. While injection devices were available in greater quantities, the proportion of prescriptions including at least one injection remained stable between 1995 (26.5%) and 2000 (23.8%). CONCLUSION: The implementation of pharmaceutical depots next to public health care facilities increased geographical access to essential medicines and basic supplies, among which syringes and needles, contributing substantially to safer injection practices in the absence of increased use of therapeutic injections. PMID- 16364179 TI - Assessment of the structure of the Hospital Anxiety and Depression Scale in musculoskeletal patients. AB - BACKGROUND: Research suggests there is a high prevalence of anxiety and depression amongst patients with chronic musculoskeletal pain, which can influence the effectiveness of rehabilitation programs. It is therefore important for clinicians involved in musculoskeletal rehabilitation programs to consider screening patients for elevated levels of anxiety and depression and to provide appropriate counselling or treatment where necessary. The HADS has been used as a screening tool for assessment of anxiety and depression in a wide variety of clinical groups. Recent research however has questioned its suitability for use with some patient groups due to problems with dimensionality and the behaviour of individual items. The aim of this study is to assess the underlying structure and psychometric properties of the HADS among patients attending musculoskeletal rehabilitation. METHODS: Data was obtained from 296 patients attending an outpatient musculoskeletal pain clinic. The total sample was used to identify the proportion of patients with elevated levels of anxiety and depression. Half the sample (n = 142) was used for exploratory factor analysis (EFA), with the holdout sample (n = 154) used for confirmatory factor analysis (CFA) to explore the underlying structure of the scale. RESULTS: A substantial proportion of patients were classified as probable cases on the HADS Anxiety subscale (38.2%) and HADS Depression subscale (30.1%), with the sample recording higher mean HADS subscales scores than many other patient groups (breast cancer, end-stage renal disease, heart disease) reported in the literature. EFA supported a two factor structure (representing anxiety and depression) as proposed by the scale's authors, however item 7 (an anxiety item) failed to load appropriately. Removing Item 7 resulted in a clear two factor solution in both EFA and CFA. CONCLUSION: The high levels of anxiety and depression detected in this sample suggests that screening for psychological comorbidity is important in musculoskeletal rehabilitation settings. It is necessary for clinicians who are considering using the HADS as a screening tool to first assess its suitability with their particular patient group. Although EFA and CFA supported the presence of two subscales representing anxiety and depression, the results with this musculoskeletal sample suggest that item 7 should be removed from the anxiety subscale. PMID- 16364181 TI - Providing education on evidence-based practice improved knowledge but did not change behaviour: a before and after study. AB - BACKGROUND: Many health professionals lack the skills to find and appraise published research. This lack of skills and associated knowledge needs to be addressed, and practice habits need to change, for evidence-based practice to occur. The aim of this before and after study was to evaluate the effect of a multifaceted intervention on the knowledge, skills, attitudes and behaviour of allied health professionals. METHODS: 114 self-selected occupational therapists were recruited. The intervention included a 2-day workshop combined with outreach support for eight months. Support involved email and telephone contact and a workplace visit. Measures were collected at baseline, post-workshop, and eight months later. The primary outcome was knowledge, measured using the Adapted Fresno Test of Evidence-Based Practice (total score 0 to 156). Secondary outcomes were attitude to evidence-based practice (% reporting improved skills and confidence; % reporting barriers), and behaviour measured using an activity diary (% engaging/not engaging in search and appraisal activities), and assignment completion. RESULTS: Post-workshop, there were significant gains in knowledge which were maintained at follow-up. The mean difference in the Adapted Fresno Test total score was 20.6 points (95% CI, 15.6 to 25.5). The change from post workshop to follow-up was small and non-significant (mean difference 1.2 points, 95% CI, -6.0 to 8.5). Fewer participants reported lack of searching and appraisal skills as barriers to evidence-based practice over time (searching = 61%, 53%, 24%; appraisal 60%, 65%, 41%). These differences were statistically significant (p = 0.0001 and 0.010 respectively). Behaviour changed little. Pre-workshop, 6% engaged in critical appraisal increasing to 18% post-workshop and 18% at follow up. Nearly two thirds (60%) were not reading any research literature at follow up. Twenty-three participants (20.2%) completed their assignment. CONCLUSION: Evidence-based practice skills and knowledge improved markedly with a targetted education intervention and outreach support. However, changes in behaviour were small, based on the frequency of searching and appraisal activities. Allied health educators should focus more on post-workshop skill development, particularly appraisal, and help learners to establish new routines and priorities around evidence-based practice. Learners also need to know that behaviour change of this nature may take months, even years. PMID- 16364180 TI - Pathophysiological classification of chronic rhinosinusitis. AB - BACKGROUND: Recent consensus statements demonstrate the breadth of the chronic rhinosinusitis (CRS) differential diagnosis. However, the classification and mechanisms of different CRS phenotypes remains problematic. METHOD: Statistical patterns of subjective and objective findings were assessed by retrospective chart review. RESULTS: CRS patients were readily divided into those with (50/99) and without (49/99) polyposis. Aspirin sensitivity was limited to 17/50 polyp subjects. They had peripheral blood eosinophilia and small airways obstruction. Allergy skin tests were positive in 71% of the remaining polyp subjects. IgE was<10 IU/ml in 8/38 polyp and 20/45 nonpolyp subjects (p = 0.015, Fisher's Exact test). CT scans of the CRS without polyp group showed sinus mucosal thickening (probable glandular hypertrophy) in 28/49, and nasal osteomeatal disease in 21/49. Immunoglobulin isotype deficiencies were more prevalent in nonpolyp than polyp subjects (p < 0.05). CONCLUSION: CRS subjects were retrospectively classified in to 4 categories using the algorithm of (1) polyp vs. nonpolyp disease, (2) aspirin sensitivity in polyposis, and (3) sinus mucosal thickening vs. nasal osteomeatal disease (CT scan extent of disease) for nonpolypoid subjects. We propose that the pathogenic mechanisms responsible for polyposis, aspirin sensitivity, humoral immunodeficiency, glandular hypertrophy, eosinophilia and atopy are primary mechanisms underlying these CRS phenotypes. The influence of microbial disease and other factors remain to be examined in this framework. We predict that future clinical studies and treatment decisions will be more logical when these interactive disease mechanisms are used to stratify CRS patients. PMID- 16364182 TI - Mechanomyographic amplitude and frequency responses during dynamic muscle actions: a comprehensive review. AB - The purpose of this review is to examine the literature that has investigated mechanomyographic (MMG) amplitude and frequency responses during dynamic muscle actions. To date, the majority of MMG research has focused on isometric muscle actions. Recent studies, however, have examined the MMG time and/or frequency domain responses during various types of dynamic activities, including dynamic constant external resistance (DCER) and isokinetic muscle actions, as well as cycle ergometry. Despite the potential influences of factors such as changes in muscle length and the thickness of the tissue between the muscle and the MMG sensor, there is convincing evidence that during dynamic muscle actions, the MMG signal provides valid information regarding muscle function. This argument is supported by consistencies in the MMG literature, such as the close relationship between MMG amplitude and power output and a linear increase in MMG amplitude with concentric torque production. There are still many issues, however, that have yet to be resolved, and the literature base for MMG during both dynamic and isometric muscle actions is far from complete. Thus, it is important to investigate the unique applications of MMG amplitude and frequency responses with different experimental designs/methodologies to continually reassess the uses/limitations of MMG. PMID- 16364184 TI - Takifugu obscurus is a euryhaline fugu species very close to Takifugu rubripes and suitable for studying osmoregulation. AB - BACKGROUND: The genome sequence of the pufferfish Takifugu rubripes is an enormously useful tool in the molecular physiology of fish. Euryhaline fish that can survive both in freshwater (FW) and seawater (SW) are also very useful for studying fish physiology, especially osmoregulation. Recently we learned that there is a pufferfish, Takifugu obscurus, common name "mefugu" that migrates into FW to spawn. If T. obscurus is indeed a euryhaline fish and shares a high sequence homology with T. rubripes, it will become a superior animal model for studying the mechanism of osmoregulation. We have therefore determined its euryhalinity and phylogenetic relationship to the members of the Takifugu family. RESULTS: The following six Takifugu species were used for the analyses: T. obscurus, T. rubripes, T. niphobles, T. pardalis, T. poecilonotus, and T. porphyreus. When transferred to FW, only T. obscurus could survive while the others could not survive more than ten days in FW. During this course of FW adaptation, serum Na+ concentration of T. obscurus decreased only slightly, but a rapid and large decrease occurred even in the case of T. niphobles, a peripheral fresh water species that is often seen in brackish river mouths. Phylogenetic analysis using nucleotide sequences of the mitochondrial 16S ribosomal RNA gene of each species indicated that the six Takifugu species are very closely related with each other. CONCLUSION: T. obscurus is capable of adapting to both FW and SW. Its genomic sequence shares a very high homology with those of the other Takifugu species such that the existing Takifugu genomic information resources can be utilized. These properties make "mefugu", which has drawn little attention from animal physiologists until this study, a useful model animal for studying the molecular mechanism of maintaining body fluid homeostasis. PMID- 16364183 TI - Self-rated health, work characteristics and health related behaviours among nurses in Greece: a cross sectional study. AB - BACKGROUND: Previous studies on self-rated health among nurses have indicated an association of low job satisfaction and stress in relation to poor self-rated health. The relationship between self rated health and the specific work characteristics and health related behaviours of nurses to our knowledge have not been adequately studied. OBJECTIVE: To investigate the health profile of nurses working in hospitals in North West Greece and to examine the associations between self rated health (SRH) and health related behaviours and work characteristics in this group of hospital employees. METHODS: A self-administered questionnaire was distributed to a random sample of 443 nurses working in all the hospitals in North West Greece. Regression analysis was used to examine the relationship of health related behaviours and work characteristics with self rated health among the nurses. RESULTS: A total of 353 responded to the questionnaire (response rate 80%) of which 311 (88%) were female and 42 (12%) male. The mean age (standard deviation) of the respondents was 36 years (5.6) and their mean years of working as nurses were 13.5 years (5.9). Almost half of the nurses' smoked, and about one third were overweight or obese. About 58% (206) of the nurses reported having poor health while 42% (147) reported having good health. Self-rated health was independently associated with gender, effort to avoid fatty foods and physical activity, according to multiple logistic regression analysis. CONCLUSION: The population studied presented a relatively poor health profile, and a high proportion of poor SRH. Though female gender and effort to avoid fatty foods were associated with poor SRH, and exercise and white meat consumption with good SRH, specific work characteristics were not associated with SRH. PMID- 16364186 TI - E-health: determinants, opportunities, challenges and the way forward for countries in the WHO African Region. AB - BACKGROUND: The implementation of the 58th World Health Assembly resolution on e health will pose a major challenge for the Member States of the World Health Organization (WHO) African Region due to lack of information and communications technology (ICT) and mass Internet connectivity, compounded by a paucity of ICT related knowledge and skills. The key objectives of this article are to: (i) explore the key determinants of personal computers (PCs), telephone mainline and cellular and Internet penetration/connectivity in the African Region; and (ii) to propose actions needed to create an enabling environment for e-health services growth and utilization in the Region. METHODS: The effects of school enrolment, per capita income and governance variables on the number of PCs, telephone mainlines, cellular phone subscribers and Internet users were estimated using a double-log regression model and cross-sectional data on various Member States in the African Region. The analysis was based on 45 of the 46 countries that comprise the Region. The data were obtained from the United Nations Development Programme (UNDP), the World Bank and the International Telecommunications Union (ITU) sources. RESULTS: There were a number of main findings: (i) the adult literacy and total number of Internet users had a statistically significant (at 5% level in a t-distribution test) positive effect on the number of PCs in a country; (ii) the combined school enrolment rate and per capita income had a statistically significant direct effect on the number of telephone mainlines and cellular telephone subscribers; (iii) the regulatory quality had statistically significant negative effect on the number of telephone mainlines; (iv) similarly, the combined school enrolment ratio and the number of telephone mainlines had a statistically significant positive relationship with Internet usage; and (v) there were major inequalities in ICT connectivity between upper-middle, lower middle and low income countries in the Region. By focusing on the adoption of specific technologies we attempted to interpret correlates in terms of relationships instead of absolute "causals". CONCLUSION: In order to improve access to health care, especially for the majority of Africans living in remote rural areas, there is need to boost the availability and utilization of e-health services. Thus, universal access to e-health ought to be a vision for all countries in the African Region. Each country ought to develop a road map in a strategic e-health plan that will, over time, enable its citizens to realize that vision. PMID- 16364185 TI - Thickness of the human cranial diploe in relation to age, sex and general body build. AB - BACKGROUND: Earlier studies have addressed the human total cranial vault thickness and generally found no correlation with sex, age or body weight. However, the thickness of the diploe has not been investigated. Our study has determined the diploeic thickness of the human cranial vault using modern autopsy material. METHODS: The diploeic bone thickness was measured in 64 individuals (43 males, 21 females) autopsied at our institute. The thickness was measured by X raying biopsies trephined at four specific locations on the skull. Complete medical records and pathologic autopsy results were available. RESULTS: There was a statistically significant difference in diploeic thickness between males and females in the frontal region only. Diploeic thickness was highly correlated with total cranial vault bone thickness, except for the left euryon in females. Subsequent analyses failed to reveal any correlations between the diploeic thickness and age and height and weight of the individual. CONCLUSION: Males overall have a thicker diploe, albeit this difference is statistically significant only in the frontal region. We could not discern any trends as pertains to diploeic thickness versus age, height or weight. Since the thickness of the diploe may be an important parameter in biomechanical modelling of the cranial vault, this means that the diploe can be built into such models based on the total cranial thickness, except for the frontal region where the sexual dimorphism must be taken into account. Our findings are consistent with previous studies relating the total cranial thickness to the same parameters, in that we found a high correlation between diploeic and total cranial thickness (except at the left euryon for females). Finally, we recommend that future studies try to incorporate CT or MR scan imaging, rather than point sampling, in order to achieve a total assessment of the dimensionalities of the diploe. PMID- 16364187 TI - T-cell signalling and immune system disorders. AB - T-cell receptor (TCR) engagement initiates intracellular signalling cascades that lead to T-cell proliferation, cytokine production and differentiation into effector cells. These cascades comprise an array of protein-tyrosine kinases, phosphatases, GTP-binding proteins and adaptor proteins that regulate generic and specialised functions. The integration of these signals is essential for the normal development, homeostasis and function of T cells. Defects in a single mediator can produce T cells that are unable to participate fully in an immune response and/or that mount an inappropriate response, which leads to immunodeficiency, autoimmunity or leukaemia/lymphomas. This review highlights some of the key players in T-cell signalling and their involvement in the development of various clinical disease states. Some of these immune-specific signalling proteins are attractive potential targets in the development of therapies to augment T-cell responses to antigen or tumours, and to treat immune cell disorders. PMID- 16364189 TI - Angiogenesis, l'enfant terrible of vascular biology is coming of age. PMID- 16364190 TI - Angiogenic and cell survival functions of vascular endothelial growth factor (VEGF). AB - Vascular endothelial growth factor (VEGF) was originally identified as an endothelial cell specific growth factor stimulating angiogenesis and vascular permeability. Some family members, VEGF C and D, are specifically involved in lymphangiogenesis. It now appears that VEGF also has autocrine functions acting as a survival factor for tumour cells protecting them from stresses such as hypoxia, chemotherapy and radiotherapy. The mechanisms of action of VEGF are still being investigated with emerging insights into overlapping pathways and cross-talk between other receptors such as the neuropilins which were not previously associated with angiogenesis. VEGF plays an important role in embryonic development and angiogenesis during wound healing and menstrual cycle in the healthy adult. VEGF is also important in a number of both malignant and non-malignant pathologies. As it plays a limited role in normal human physiology, VEGF is an attractive therapeutic target in diseases where VEGF plays a key role. It was originally thought that in pathological conditions such as cancer, VEGF functioned solely as an angiogenic factor, stimulating new vessel formation and increasing vascular permeability. It has since emerged it plays a multifunctional role where it can also have autocrine pro-survival effects and contribute to tumour cell chemoresistance. In this review we discuss the established role of VEGF in angiogenesis and the underlying mechanisms. We discuss its role as a survival factor and mechanisms whereby angiogenesis inhibition improves efficacy of chemotherapy regimes. Finally, we discuss the therapeutic implications of targeting angiogenesis and VEGF receptors, particularly in cancer therapy. PMID- 16364191 TI - Metabolic state signalling through central hypocretin/orexin neurons. AB - Hypothalamic neurons that produce the peptide transmitters hypocretins/orexins have attracted much recent attention. They provide direct and predominantly excitatory inputs to all major brain areas except the cerebellum, with the net effect of stimulating wakefulness and arousal. These inputs are essential for generating sustained wakefulness in mammals, and defects in hypocretin signalling result in narcolepsy. In addition, new roles for hypocretins/orexins are emerging in reward-seeking, learning, and memory. Recent studies also indicate that hypocretin/orexin neurons can alter their intrinsic electrical activity according to ambient fluctuations in the levels of nutrients and appetite-regulating hormones. These intriguing electrical responses are perhaps the strongest candidates to date for the elusive neural correlates of after-meal sleepiness and hunger-induced wakefulness. Hypocretin/orexin neurons may thus directly translate rises and falls in body energy levels into different states of consciousness. PMID- 16364192 TI - Embryonic stem cells and cardiomyocyte differentiation: phenotypic and molecular analyses. AB - Embryonic stem (ES) cell lines, derived from the inner cell mass (ICM) of blastocyst-stage embryos, are pluripotent and have a virtually unlimited capacity for self-renewal and differentiation into all cell types of an embryoproper. Both human and mouse ES cell lines are the subject of intensive investigation for potential applications in developmental biology and medicine. ES cells from both sources differentiate in vitro into cells of ecto-, endoand meso-dermal lineages, and robust cardiomyogenic differentiation is readily observed in spontaneously differentiating ES cells when cultured under appropriate conditions. Molecular, cellular and physiologic analyses demonstrate that ES cell-derived cardiomyocytes are functionally viable and that these cell derivatives exhibit characteristics typical of heart cells in early stages of cardiac development. Because terminal heart failure is characterized by a significant loss of cardiomyocytes, the use of human ES cell-derived progeny represents one possible source for cell transplantation therapies. With these issues in mind, this review will focus on the differentiation of pluripotent embryonic stem cells into cardiomyocytes as a developmental model, and the possible use of ES cell-derived cardiomyocytes as source of donor cells. PMID- 16364193 TI - Pharmacological basis of different targets for the treatment of atherosclerosis. AB - The development of atherosclerotic plaque is a highly regulated and complex process which occurs as a result of structural and functional alterations in endothelial cells, smooth muscle cells (SMCs), monocytes/macrophages, T lymphocytes and platelets. The plaque formation in the coronary arteries or rupture of the plaque in the peripheral vasculature in latter stages of atherosclerosis triggers the onset of acute ischemic events involving myocardium. Although lipid lowering with statins has been established as an important therapy for the treatment of atherosclerosis, partially beneficial effects of statins beyond decreasing lipid levels has shifted the focus to develop newer drugs that can affect directly the process of atherosclerosis. Blockade of renin angiotensin system, augmentation of nitric oxide availability, reduction of Ca(2+) influx, prevention of oxidative stress as well as attenuation of inflammation, platelet activation and SMC proliferation have been recognized as targets for drug treatment to control the development, progression and management of atherosclerosis. A major challenge for future drug development is to formulate a combination therapy affecting different targets to improve the treatment of atherosclerosis. PMID- 16364194 TI - Antisense- and RNA interference-based therapeutic strategies in allergy. AB - Modern therapeutic methods for manipulation of gene expression in allergic diseases have been receiving increased attention in the emerging era of functional genomics. With the growing application of gene silencing technologies, pharmacological modulation of translation represents a great advance in molecular therapy for allergy. Several strategies for sequence-specific post transcriptional inhibition of gene expression can be distinguished: antisense oligonucleotides (AS-ONs), ribozymes (RZs), DNA enzymes (DNAzymes), and RNA interference (RNAi) triggered by small interfering RNAs (siRNAs). Potential anti mRNA drugs in asthma and other allergic disorders may be targeted to cell surface receptors (adenosine A1 receptor, high-affinity receptor Fc-epsilon RI-alpha, cytokine receptors), adhesion molecules and ligands (ICAM-1, VLA-4), ion channels (calcium-dependent chloride channel-1), cytokines and related factors (IL-4, IL 5, IL-13, SCF, TNF-alpha, TGF-beta1), intracellular signal transduction molecules, such as tyrosine-protein kinases (Syk, Lyn, Btk), serine/ threonine protein kinases (p38 alpha MAPkinase, Raf-1), non-kinase signaling proteins (RasGRP4), and transcription factors involved in Th2 differentiation and allergic inflammation (STAT-6, GATA-3, NF-kappaB). The challenge to scientists is to determine which of the candidate targets warrants investment of time and resources. New-generation respirable AS-ONs, external guide sequence ribozymes, and RNA interference-based therapies have the potential to satisfy unmet needs in allergy treatment, acting at a more proximal level to a key etiopathogenetic molecular process, represented by abnormal expression of genes. Moreover, antisense and siRNA technologies imply a more rational design of new drugs for allergy. PMID- 16364195 TI - A comprehensive view of polyamine and histamine metabolism to the light of new technologies. AB - Polyamines and histamine are biogenic amines with multiple biological roles. In spite of the evidence for the involvement of both polyamines and histamine metabolism impairment in several highly prevalent pathological conditions, multiple questions concerning the molecular processes behind these effects remain to be elucidated. More comprehensive and systemic studies integrating molecular biology, biophysical and bioinformatics tools could contribute to accelerate the advances in this research area. This review is designed to underscore the main questions to be answered in polyamine and histamine research and how these new systemic approaches could help to find these answers. PMID- 16364196 TI - Animal disease models generated by genetic engineering of polyamine metabolism. AB - The polyamines putrescine, spermidine and spermine are natural components of all living cells. Although their exact cellular functions are still largely unknown, a constant supply of these compounds is required for mammalian cell proliferation to occur. Studies with animals displaying genetically altered polyamine metabolism have shown that polyamines are intimately involved in the development of diverse tumors, putrescine apparently has specific role in skin physiology and neuroprotection and the higher polyamines spermidine and spermine are required for the maintenance of pancreatic integrity and liver regeneration. In the absence of ongoing polyamine biosynthesis, murine embryogenesis does not proceed beyond the blastocyst stage. The last years have also witnessed the appearance of the first reports linking genetically altered polyamine metabolism to human diseases. PMID- 16364197 TI - Impact of electrical stimulation on three-dimensional myoblast cultures - a real time RT-PCR study. AB - Several focal skeletal muscle diseases, including tumours and trauma lead to a limited loss of functional muscle tissue. There is still no suitable clinical approach for treating such defects. A promising approach could be the tissue engineering of skeletal muscle. However, a clinically reliable differentiation stimulus for three-dimensional (3-D) cultures is necessary for this process, and this condition has not yet been established. In order to quantify and analyze the differentiation potential of electrical cell stimulation, primary myoblasts were stimulated within a 3-D fibrin- matrix. Gene expression of MyoD, myogenin and AChR-epsilon were measured by real-time RT-PCR over a time period of eight days, showing immediate down-regulation of all marker genes. For tissue engineering approaches, cell multiplication is crucial for acquisition of sufficient tissue volumes for reconstruction. Therefore, all experiments were performed with high and low passaged myoblasts, demonstrating higher transcript rates of marker genes in lowpassage cells. Our findings strongly suggest a reconsideration of electrical stimulation in muscle tissue engineering. PMID- 16364198 TI - Interstitial Cajal-like cells (ICLC) in human resting mammary gland stroma. Transmission electron microscope (TEM) identification. AB - We have previously shown the existence of ICLC in human resting mammary gland stroma by means of methylene blue (vital) staining and c-kit immunopositivity (immunofluorescence and immunohistochemistry). In addition, we reported the phenotype characteristics of these ICLC in vitro (primary cell cultures). Since the identification of ICLC outside the gut requires, at this moment, the obligatory use of TEM, we used this technique and provide unequivocal evidence for the presence of ICLC in the intralobular stroma of human resting mammary gland. According to the 'platinum standard' (10 TEM criteria for the certitude diagnosis of ICLC), we found interstitial cells with the following characteristics: 1. location: among the tubulo-alveolar structures, in the non epithelial space; 2. caveolae: approximately 2.5% of cell volume; 3. mitochondria: approximately 10% of cell volume; 4. endoplasmic reticulum: either smooth or rough, approximately 2-3% of cell volume; 5. cytoskeleton: intermediate and thin filaments, as well as microtubules are present; 6. myosin thick filaments: undetectable; 7. basal lamina: occasionally found; 8. gap junctions: occasionally found; 9. close contacts with targets: nerve fibers, capillaries, immunoreactive cells by 'stromal synapses'; 10. characteristic cytoplasmic processes: i) number: frequently 2-3; ii) length: several tens of mum; iii) thickness: uneven caliber, 0.1-0.5 microm, with dilations, but very thin from the emerging point; iv) aspect: moniliform, usually with mitochondria located in dilations; v) branching: dichotomous pattern; vi) Ca(2+) release units: are present; vii) network labyrinthic system: overlapping cytoplasmic processes. It remains to be established which of the possible roles that we previously suggested for ICLC (e.g. juxta- and/or paracrine secretion, uncommited progenitor cells, immunological surveillance, intercellular signaling, etc.) are essential for the epithelium/stroma equilibrium in the mammary gland under normal or pathological conditions. PMID- 16364199 TI - Bone induction by recombinant human osteogenic protein-1 (hOP-1, BMP-7) in the primate Papio ursinus with expression of mRNA of gene products of the TGF-beta superfamily. AB - Predictable bone induction in clinical contexts requires information on the expression and cross regulation of gene products of the transforming growth factor-beta (TGF-beta) superfamily elicited by single applications of each recombinant human bone morphogenetic/osteogenic proteins (BMPs/OPs). Using the calvarium and the rectus abdominis muscle of adult baboons Papio ursinus as a model for tissue induction and morphogenesis, this study investigated the induction of bone morphogenesis by gamma-irradiated hOP-1 delivered by gamma irradiated bovine insoluble collagenous bone matrix, the hOP-1 osteogenic device, for bone induction in heterotopic and orthotopic sites of the primate Papio ursinus and the expression patterns of OP-1, collagen type IV, BMP-3 and TGFbeta1mRNAs elicited by increasing single applications of doses of the hOP-1 osteogenic devices (0.1, 0.5 and 2.5 mg hOP-1/g of matrix) applied heterotopically in the rectus abdominis muscle and orthotopically in 48 calvarial defects of 12 adult baboons. Histology and histomorphometry on serial undecalcified sections prepared from the specimens harvested on day 15, 30 and 90 showed that all the doses of the hOP-1 osteogenic device induced bone formation culminating in complete calvarial regeneration by day 90. Type IV collagen mRNA expression, a marker of angiogenesis, was strongly expressed in both heterotopic and orthotopic tissues. High levels of expression of OP-1 mRNA demonstrated autoinduction of OP-1 mRNAs. Expression levels of BMP-3 mRNA varied from tissues induced in heterotopic vs. orthotopic sites with high expression in rapidly forming heterotopic ossicles together with high expression of type IV collagen mRNA. The temporal and spatial expressions of TGF-beta1 mRNAindicate a specific temporal transcriptional window during which expression of TGF-beta1 is mandatory for successful and optimal osteogenesis. The induction of bone by hOP-1 in Papio ursinus develops as a mosaic structure with distinct spatial and temporal patterns of gene expression of members of the TGF-beta superfamily that singly, synergistically and synchronously initiate and maintain tissue induction and morphogenesis. PMID- 16364200 TI - Molecular and cellular characterization during chondrogenic differentiation of adipose tissue-derived stromal cells in vitro and cartilage formation in vivo. AB - Human adipose tissue is a viable source of mesenchymal stem cells (MSCs) with wide differentiation potential for musculoskeletal tissue engineering research. The stem cell population, termed processed lipoaspirate (PLA) cells, can be isolated from human lipoaspirates and expanded in vitro easily. This study was to determine molecular and cellular characterization of PLA cells during chondrogenic differentiation in vitro and cartilage formation in vivo. When cultured in vitro with chondrogenic medium as monolayers in high density, they could be induced toward the chondrogenic lineages. To determine their ability of cartilage formation in vivo, the induced cells in alginate gel were implanted in nude mice subcutaneously for up to 20 weeks. Histological and immunohistochemical analysis of the induced cells and retrieved specimens from nude mice at various intervals showed obviously cartilaginous phenotype with positive staining of specific extracellular matrix (ECM). Correlatively, results of RT-PCR and Western Blot confirmed the expression of characteristic molecules during chondrogenic differentiation namely collagen type II, SOX9, cartilage oligomeric protein (COMP) and the cartilage-specific proteoglycan aggrecan. Meanwhile, there was low level synthesis of collagen type X and decreasing production of collagen type I during induction in vitro and formation of cartilaginous tissue in vivo. These cells induced to form engineered cartilage can maintain the stable phenotype and indicate no sign of hypertrophy in 20 weeks in vivo, however, when they cultured as monolayers, they showed prehypertrophic alteration in late stage about 10 weeks after induction. Therefore, it is suggested that human adipose tissue may represent a novel plentiful source of multipotential stem cells capable of undergoing chondrogenesis and forming engineered cartilage. PMID- 16364201 TI - Modulation of adenylyl cyclase activity in young and adult rat brain cortex. Identification of suramin as a direct inhibitor of adenylyl cyclase. AB - Adenylyl cyclase (AC) in brain cortex from young (12-day-old) rats exhibits markedly higher activity than in adult (90-day-old) animals. In order to find some possibly different regulatory features of AC in these two age groups, here we modulated AC activity by dithiothreitol (DTT), Fe(2+), ascorbic acid and suramin. We did not detect any substantial difference between the effects of all these tested agents on AC activity in cerebrocortical membranes from young and adult rats, and the enzyme activity was always about two-fold higher in the former preparations. Nevertheless, several interesting findings have come out of these investigations. Whereas forskolin- and Mn(2+)-stimulated AC activity was significantly enhanced by the addition of DTT, increased concentrations of Fe(2+) ions or ascorbic acid substantially suppressed the enzyme activity. Lipid peroxidation induced by suitable combinations of DTT/Fe(2+) or by ascorbic acid did not influence AC activity. We have also observed that PKC- or protein tyrosine kinase-mediated phosphorylation apparently does not play any significant role in different activity of AC determined in cerebrocortical preparations from young and adult rats. Our experiments analysing the presumed modulatory role of suramin revealed that this pharmacologically important drug may act as a direct inhibitor of AC. The enzyme activity was diminished to the same extent by suramin in membranes from both tested age groups. Our present data show that AC is regulated similarly in brain cortex from both young and adult rats, but its overall activity is much lower in adulthood. PMID- 16364202 TI - Inhibitory effect of acetylsalicylic acid on matrix metalloproteinase - 2 activity in human endothelial cells exposed to high glucose. AB - Matrix metalloproteinases play a major role in the process of angiogenesis, an important feature of diabetes complications, cancer or rheumatoid arthritis. High glucose concentrations were reported to augment metalloproteinase-2 secretion in some cell types. In the present study we investigated the influence of acetylsalicylic acid on metalloproteinase- 2 secretion and expression in endothelial cells cultured for one week in high glucose conditions (25 mM and 33 mM). Metalloproteinase-2 activity was evidenced by gel zymography, the protein was identified by Western blotting, and the gene expression was quantitated by RT PCR. The results indicated a marked inhibitory effect of acetylsalicylic acid at gene expression level (approximately 43%) and also at secretion level in samples of conditioned media (approximately 30%) and cellular homogenates (approximately 70%). This may suggest that acetylsalicylic acid could have a beneficial effect in preventing the angiogenic process that appears in diabetes complications. PMID- 16364203 TI - Evaluation of the ELOVL4 gene in a Chinese family with autosomal dominant STGD3 like macular dystrophy. AB - Stargardt disease-3 (STGD3) is an autosomal dominant juvenile-onset macular dystrophy characterized by progressive decreasing visual acuity, bilateral atrophic changes in the macula and absence of characteristic dark choroids. We identified a STGD3-like macular dystrophy pedigree by clinical examination. To explore whether the STGD3-like phenotype in the kindred is linked to ELOVL4 gene or associated with any other identified STGD gene, we extracted genomic DNA from leukocytes of peripheral blood from the available family members and 50 normal controls for mutation analysis. Then the exons of ELOVL4, RDS and the three exons of ABCR were amplified by polymerase chain reaction (PCR). All PCR products were screened for mutations by combination of denaturing high-performance liquid chromatography (DHPLC) analysis and DNA sequencing. No mutation was found in the exons of three candidate genes, but we obtained three non-pathogenic polymorphisms, IVS5-2533T-->A in ELOVL4, 558C-->T (Val106Val) and 1150G-->C (Glu304Gln) in RDS. And IVS5-2533T-->A is never shown in the previous references. These data suggested that there exist other unknown genes responsible for the STGD3-like phenotype in the pedigree. PMID- 16364204 TI - Amlodipine preserves the glomerular number in spontaneously hypertensive rats. AB - The calcium channel blockers have individual pharmacological and therapeutic properties that may vary, but as a group, they are effective antihypertensive agents in patients with renal disease. Their effects on the kidney may extend beyond BP reduction alone. Fifteen one-year-old male spontaneously hypertensive rats (SHR) were separated in three groups: Initial control group (IC), Final control group (FC, SHR received standard rat chow and fresh water ad libitum during 15 weeks), Amlodipine group (Aml, SHR) received 0.2 mg/kg/day of amlodipine in addition to food and water during 15 weeks. The glomerular number was estimated using the disector method. In the Control group, the BP level increased almost 20 per cent in the first six weeks (from 186 +/- 11 to 223 +/- 16 mmHg, p<0.01) and then BP level increased almost 15 percent until week 15 (from 223 +/- 16 to 258 +/- 20 mmHg, p<0.01). In the same period, the Aml group showed a progressively low BP, reaching a level almost 50 per cent lower in the week 15 than in the week 1 (from 190 +/- 15 to 101+/-8 mmHg, p<0.01). Amlodipine treatment significantly decreased the serum creatinine, more than 12 per cent lower than the FC group (from 70.4 +/- 6.2 to 61.4 +/- 5.2 micromol/L, p<0.05). However, proteinuria was not different when groups were compared. The FC group reached a glomerular number almost 20 percent smaller than the IC and Aml groups (from 35 x 10(3) +/- 7 x 10(3) in IC group, 34 x 10(3) +/- 4 x 10(3) in Aml group to 27 x 10(3) +/- 3 x 10(3) in FC group, p<0.05). A possible protective effect of amlodipine against the loss of glomeruli in SHR is a major additional action of amlodipine in the treatment of hypertension mainly when the renal lesion already exists. PMID- 16364206 TI - The telomere length dynamic and methods of its assessment. AB - Human telomeres are composed of long repeating sequences of TTAGGG, associated with a variety of telomere-binding proteins. Its function as an end-protector of chromosomes prevents the chromosome from end-to-end fusion, recombination and degradation. Telomerase acts as reverse transcriptase in the elongation of telomeres, which prevent the loss of telomeres due to the end replication problems. However, telomerase activity is detected at low level in somatic cells and high level in embryonic stem cells and tumor cells. It confers immortality to embryonic stem cells and tumor cells. In most tumor cells, telomeres are extremely short and stable. Telomere length is an important indicator of the telomerase activity in tumor cells and it may be used in the prognosis of malignancy. Thus, the assessment of telomeres length is of great experimental and clinical significance. This review describes the role of telomere and telomerase in cancer pathogenesis and the dynamics of the telomeres length in different cell types. The various methods of measurement of telomeres length, i.e. southern blot, hybridization protection assay, fluorescence in situ hybridization, primed in situ, quantitative PCR and single telomere length analysis are discussed. The principle and comparative evaluation of these methods are reviewed. The detection of G-strand overhang by telomeric-oligonucleotide ligation assay, primer extension/nick translation assay and electron microscopy are briefly discussed. PMID- 16364205 TI - Interstitial Cajal-like cells (ICLC) in human atrial myocardium. AB - We present here visual evidence for the existence of a new type of interstitial cells in human atrial myocardium: interstitial Cajal-like cells (ICLC). These cells fulfil the so-called 'platinum standard' (a set of 10 ultrastructural criteria for the positive diagnosis of ICLC). Conventional transmission electron microscopy (TEM), followed by reconstructions from serial photomicrographs, revealed typical ICLC with 2 or 3 long, moniliform processes (several tens of micrometers long and 0.1-0.5 microm thick), emerging from the (small) cell body. Cell processes dichotomously branch and have mitochondria (at the level of dilations), caveolae and Ca(2+) release units. Cell prolongations establish close spatial relationships between each other, as well as with capillaries, myocardial cells, and other connective tissue cells. Our preliminary data suggest that ICLC exist in rat ventricular myocardium, too. PMID- 16364207 TI - Progress in studies of huperzine A, a natural cholinesterase inhibitor from Chinese herbal medicine. AB - Huperzine A (HupA), a novel alkaloid isolated from the Chinese herb Huperzia serrata, is a potent, highly specific and reversible inhibitor of acetylcholinesterase(AChE). Compared with tacrine, donepezil, and rivastigmine, HupA has better penetration through the blood-brain barrier, higher oral bioavailability, and longer duration of AChE inhibitory action. HupA has been found to improve cognitive deficits in a broad range of animal models. HupA possesses the ability to protect cells against hydrogen peroxide, beta-amyloid protein (or peptide), glutamate, ischemia and staurosporine-induced cytotoxicity and apoptosis. These protective effects are related to its ability to attenuate oxidative stress, regulate the expression of apoptotic proteins Bcl-2, Bax, P53, and caspase-3, protect mitochondria, upregulate nerve growth factor and its receptors, and interfere with amyloid precursor protein metabolism. Antagonizing effects of HupA on N-methyl-D-aspartate receptors and potassium currents may also contribute to its neuroprotection as well. Pharmacokinetic studies in rodents, canines, and healthy human volunteers indicated that HupA was absorbed rapidly, distributed widely in the body, and eliminated at a moderate rate with the property of slow and prolonged release after oral administration. Animal and clinical safety tests showed that HupA had no unexpected toxicity, particularly the dose-limiting hepatotoxicity induced by tacrine. The phase IV clinical trials in China have demonstrated that HupA significantly improved memory deficits in elderly people with benign senescent forgetfulness, and patients with Alzheimer disease and vascular dementia, with minimal peripheral cholinergic side effects and no unexpected toxicity. HupA can also be used as a protective agent against organophosphate intoxication. PMID- 16364208 TI - Flavonoid-membrane interactions: possible consequences for biological effects of some polyphenolic compounds. AB - Flavonoids are found ubiquitously in higher plants and constitute an important component of the majority of people daily diets. The biological activities of flavonoids cover a very broad spectrum, from anticancer and antibacterial activities through to inhibition of bone resorption. In the present paper, the interactions between flavonoids and lipid bilayers as well as biological membranes and their components are reviewed, with special emphasis on the structure-activity relationships and mechanisms underlying the biological activity of flavonoids. PMID- 16364209 TI - Role of melatonin in Alzheimer-like neurodegeneration. AB - Alzheimer disease (AD), an age-related neurodegenerative disorder with progressive loss of memory and deterioration of comprehensive cognition, is characterized by extracellular senile plaques of aggregated beta-amyloid (Abeta), and intracellular neurofibrillary tangles that contain hyperphosphorylated tau protein. Recent studies showed that melatonin, an indoleamine secreted by the pineal gland, may play an important role in aging and AD as an antioxidant and neuroprotector. Melatonin decreases during aging and patients with AD have a more profound reduction in this hormone. Data from clinical trials indicate that melatonin supplementation improves sleep, ameliorates sundowning, and slows down the progression of cognitive impairment in Alzheimer patients. Melatonin efficiently protects neuronal cells from Abeta-mediated toxicity via antioxidant and anti-amyloid properties: it not only inhibits Abeta generation, but also arrests the formation of amyloid fibrils by a structure-dependent interaction with Abeta. Our recent studies have demonstrated that melatonin efficiently attenuates Alzheimer-like tau hyperphosphorylation. Although the exact mechanism is still not fully understood, a direct regulatory influence of melatonin on the activities of protein kinases and protein phosphatases is proposed. Additionally, melatonin also plays a role in protecting cholinergic neurons and in anti inflammation. Here, the neuroprotective effects of melatonin and the underlying mechanisms by which it exerts its effects are reviewed. The capacity of melatonin to prevent or ameliorate tau and Abeta pathology further enhances its potential in the prevention or treatment of AD. PMID- 16364210 TI - Growth-promoting activity of Hominis Placenta extract on regenerating sciatic nerve. AB - AIM: Extract of Hominis Placenta (HP) has been used in oriental medicine as an agent for improving physiological function. The present study was conducted to investigate whether HP treatment in an experimental sciatic nerve injury animal model produces growth-promoting effects on regenerating peripheral nerve fibers after injury. METHODS: After HP was injected into a sciatic nerve injury site, changes in protein levels were analyzed in the regenerating nerve area by Western blotting and immunofluorescence staining analyses. For quantitative assessment of axonal regeneration, a retrograde tracing technique was used to identify the neuronal cell bodies corresponding to regenerating axons, and the extent of neurite outgrowth in cultured dorsal root ganglia (DRG) sensory neurons prepared from animals that had experienced a sciatic nerve crush injury 7 d before neuron collection was analyzed. RESULTS: Induction levels of axonal growth-associated protein (GAP-43) in the injured sciatic nerves were elevated by HP treatment. HP treatment also upregulated cell division cycle 2 (Cdc2) protein levels in the distal stump of the injured sciatic nerve. Induced Cdc2 protein was detected in Schwann cells, suggesting that Cdc2 kinase activity may be involved in the growth promoting activity of regenerating axons via Schwann cell proliferation. Cell body measurement by retrograde tracing indicated that HP treatment produced significant increases in regenerating motor axons. Finally, HP treatment of cultured DRG sensory neurons significantly increased neurite arborization and elongation. CONCLUSION: HP promotes the regeneration of injured sciatic axons by upregulating the synthesis of regeneration-related protein factors such as GAP-43 and Cdc2. PMID- 16364211 TI - Soybean isoflavones alter parvalbumin in hippocampus of mid-aged normal female, ovariectomized female, and normal male rats. AB - AIM: To investigate the long-term effect of soybean isoflavones on changes in parvalbumin (PV) immunoreactivity in the hippocampus in normal female, ovariectomized (OVX) female and normal male rats. METHODS: Ten-month-old rats were assigned to one of 9 groups (n = 7 in each group) based on body weight using a randomized complete-block design. The groups were: control diet-treated females, OVX females, and males; 0.3 g/kg isoflavone-treated females, OVX females, and males; and 1.2 g/kg isoflavone-treated females, OVX females, and males. The PV immunostaining was conducted by using the standard avidin-biotin complex method. RESULTS: PV immunoreactivity and the number of PV-immunoreactive neurons in all the groups after isoflavone treatment were significantly changed in the hippocampal CA1 region and in the dentate gyrus, but not in the hippocampal CA2/3 region. PV immunoreactivity and the number of PV-immunoreactive neurons in the control diet OVX females were similar to those in the control diet, and were greater than those in the control diet normal females. PV immunoreactivity and the number of PV-immunoreactive neurons in all the isoflavone-treated groups decreased dose-dependently after isoflavone treatment. CONCLUSION: Long-term administration of isoflavones may induce a reduction of PV in interneurons in the hippocampal CA1 region and in the dentate gyrus. The reduction of PV in these regions suggests that the long-term administration of isoflavones may cause a change in calcium homeostasis in the hippocampal CA1 region and in the dentate gyrus. PMID- 16364212 TI - Toosendanin interferes with pore formation of botulinum toxin type A in PC12 cell membrane. AB - AIM: Botulinum neurotoxins (BoNT) abort the process of neurotransmitter release at presynaptic motor nerve terminals, causing muscle paralysis. The ability of botulinum toxin to produce its effect is dependent on the ability of the light chain to cleave the SNARE proteins associated with transmitter release. Translocation of the light chain protease through the heavy chain-formed channel is a pivotal step in the intoxication process. Toosendanin (TSN), a triterpenoid derivative extracted from a Chinese traditional medicine, has been demonstrated to be an effective cure for experimental botulism. This study was designed to explore the antibotulismic mechanisms of toosendanin. METHODS: The inside-out single-channel recording patch-clamp technique was used to record the BoNT/A induced currents in the presence and absence of TSN. RESULTS: Channel formation was delayed and the sizes of the channels were reduced in the TSN-treated PC12 cell membrane. CONCLUSION: The antibotulismic effect of TSN might occur via interference with toxin translocation. PMID- 16364213 TI - Arecoline inhibits catecholamine release from perfused rat adrenal gland. AB - AIM: To study the effect of arecoline, an alkaloid isolated from Areca catechu, on the secretion of catecholamines (CA) evoked by cholinergic agonists and the membrane depolarizer from isolated perfused rat adrenal gland. METHODS: Adrenal glands were isolated from male Sprague-Dawley rats. The adrenal glands were perfused with Krebs bicarbonate solution by means of a peristaltic pump. The CA content of the perfusate was measured directly using the fluorometric method. RESULTS: Arecoline (0.1-1.0 mmol/L) perfused into an adrenal vein for 60 min produced dose- and time-dependent inhibition in CA secretory responses evoked by acetylcholine (ACh) (5.32 mmol/L), 1.1-dimethyl-4-phenyl piperazinium iodide (DMPP) (100 micromol/L for 2 min) and 3-(m-choloro-phenyl-carbamoyl-oxy)-2 butynyl trimethyl ammonium chloride (McN-A-343) (100 micromol/L for 2 min). However, lower doses of arecoline did not affect CA secretion of high K(+) (56 mmol/L); higher doses greatly reduced CA secretion of high K(+). Arecoline also failed to affect basal catecholamine output. Furthermore, in adrenal glands loaded with arecoline (0.3 mmol/L), CA secretory response evoked by Bay-K-8644 (10 micromol/L), an activator of L-type Ca(2+) channels, was markedly inhibited, whereas CA secretion by cyclopiazonic acid (10 micromol/L), an inhibitor of cytoplasmic Ca(2+)-ATPase, was not affected. Nicotine (30 micromol/L), which was perfused into the adrenal gland for 60 min, however, initially enhanced ACh evoked CA secretory responses. As time elapsed, these responses became more inhibited, whereas the initially enhanced high K(+)-evoked CA release diminished. CA secretion evoked by DMPP and McN-A-343 was significantly depressed in the presence of nicotine. CONCLUSION: Arecoline dose-dependently inhibits CA secretion from isolated perfused rat adrenal gland evoked by activation of cholinergic receptors. At lower doses arecoline does not inhibit CA secretion through membrane depolarization, but at larger doses it does. This inhibitory effect of arecoline may be mediated by blocking the calcium influx into the rat adrenal medullary chromaffin cells without the inhibition of Ca(2+) release from the cytoplasmic calcium store. There seems to be a difference in the mode of action of nicotine and arecoline in rat adrenomedullary CA secretion. PMID- 16364214 TI - Effect of leukemia inhibitory factor on embryonic stem cell differentiation: implications for supporting neuronal differentiation. AB - AIM: Leukemia inhibitory factor (LIF), a pleiotropic cytokine, has been used extensively in the maintenance of mouse embryonic stem cell pluripotency. In this current work, we examined the effect of the LIF signaling pathway in embryonic stem (ES) cell differentiation to a neural fate. METHODS: In the presence of LIF (1000 U/mL), the production of neuronal cells derived from embryoid bodies (EB) was tested under various culture conditions. Inhibition of the LIF pathway was examined with specific inhibitors. The effects of cell apoptosis and proliferation on neural differentiation were examined. ES cell differentiation into three-germ layers was compared. RESULTS: Under various culture conditions, neuronal differentiation was increased in the presence of LIF. Blocking the LIF activated STAT3 signaling pathway with specific inhibitors abolished the neuronal differentiation of ES cells, whereas inhibition of the LIF-activated MEK signaling pathway impaired the differentiation of ES cells toward a glial fate. LIF suppressed cell apoptosis and promoted cell proliferation during ES cell differentiation. LIF inhibited the differentiation of ES cells to both mesoderm and extraembryonic endoderm fates, but enhanced the determination of neural progenitors. CONCLUSION: These results suggest that LIF plays a positive role during the differentiation of ES cells into neuronal cells. PMID- 16364215 TI - Display of aggregation-prone ligand binding domain of human PPAR gamma on surface of bacteriophage lambda. AB - AIM: To display the aggregation-prone ligand binding domain (LBD) of the human peroxisome proliferator-activated receptor gamma (PPARgamma) on the surface of bacteriophages to establish an easy screening assay for the identification of PPARgamma ligands. METHODS: Plasmids were constructed for the expression of the PPARgamma LBD as a fusion to the N-terminus of the g3p protein of filamentous phage or the C-terminus of the capsid protein D (pD) of phage lambda. The fusion proteins were expressed in E coli and solubility characteristics were compared. Polyclonal antibodies against the LBD as well as the pD protein were prepared for Western blot analysis and phage capture assay. RESULTS: The pD-LBD fusion protein was partially soluble, whereas the LBD-g3p fusion protein was detected only in the insoluble fraction. The pD-LBD fusion protein was efficiently incorporated in phage particles. Furthermore, the LBD was shown to be displayed on the surface of bacteriophage lambda. On average, the pD-LBD fusion protein accounted for 28% of the total pD protein in the lambda head capsid. CONCLUSION: The hydrophobic PPARgamma LBD was expressed as a soluble form of fusion protein in E coli and displayed on the surface of bacteriophage lambda when it was fused to the lambda pD protein. The lambda pD fusion system could be used for improving the solubility of proteins that tend to form inclusion bodies when expressed in E coli. The lambda phage particles displaying the LBD of PPARgamma may be of great value for the identification of novel PPARgamma ligands. PMID- 16364216 TI - Molecular dynamics simulations of interaction between protein-tyrosine phosphatase 1B and a bidentate inhibitor. AB - AIM: To investigate the dynamic properties of protein-tyrosine phosphatase (PTP) 1B and reveal the structural factors responsible for the high inhibitory potency and selectivity of the inhibitor SNA for PTP1B. METHODS: We performed molecular dynamics (MD) simulations using a long time-scale for both PTP1B and PTP1B complexed with the inhibitor SNA, the most potent and selective PTP1B inhibitor reported to date. The trajectories were analyzed by using principal component analysis. RESULTS: Trajectory analyses showed that upon binding the ligand, the flexibility of the entire PTP1B molecule decreases. The most notable change is the movement of the WPD-loop. Our simulation results also indicated that electrostatic interactions contribute more to PTP1B-SNA complex conformation than the van der Waals interactions, and that Lys41, Arg47, and Asp48 play important roles in determining the conformation of the inhibitor SNA and in the potency and selectivity of the inhibitor. Of these, Arg47 contributed most. These results were in agreement with previous experimental results. CONCLUSION: The information presented here suggests that potent and selective PTP1B inhibitors can be designed by targeting the surface residues, for example the region containing Lys41, Arg47, and Asp48, instead of the second phosphate binding site (besides the active phosphate binding site). PMID- 16364217 TI - Interleukin-1 receptor antagonist intervenes in signaling between different types of synoviocytes in rats with adjuvant arthritis. AB - AIM: To investigate the mechanisms of interleukin-1 receptor antagonist (IL-1ra) in the treatment of adjuvant arthritis (AA). METHODS: AA was induced in rats by treatment with Freunds complete adjuvant (FCA). Rats were given an intracutaneous injection of IL-1ra (2.5, 10, 40 mg/kg, 3 times per day) from d 14 to d 21 after immunization. Synoviocyte proliferation and the activity of IL-1 were determined by using MTT assay. Tumor necrosis factor alpha (TNF-alpha) and prostaglandin E(2) (PGE(2)) concentrations were measured by radioimmunoassay. The ultrastructure of synoviocytes was observed by using a transmission electron microscope. Phosphorylation of c-Jun N-terminal kinase (JNK), extracellular regulating kinase (ERK) and p38 kinase were detected by Western blot analysis. RESULTS: IL-1ra (10 and 40 mg/kg, ic, d 14-21) modulated the secondary inflammatory reaction (P < 0.01), ultrastructure of synoviocytes and mitogen activated protein kinase (MAPK) phosphorylation in AA rats. The administration of IL-1ra (10 and 40 mg/kg, ic, d 14-21) in AA rats significantly decreased the production of IL-1, PGE2 and TNF-alpha by macrophage-like synoviocytes (MLS) (P < 0.01). IL-1ra (2.5 mg/kg) also decreased the production of PGE2 (P < 0.01) and TNF-alpha (P < 0.05) by MLS in AA rats. The increased phosphorylation of MAPK and cell proliferation in fibroblast-like synoviocytes (FLS) stimulated by supernatants of MLS in AA rats was also inhibited by IL-1ra (10 and 40 mg/kg, ic, d 14-21). CONCLUSION: IL-1ra has anti-inflammatory effects because it modulates the ultrastructure of synoviocytes, decreases the production of pro-inflammatory mediators by MLS, and inhibits the phosphorylation of MAPK in FLS. PMID- 16364218 TI - Altered pharmacokinetics of zalcitabine by concurrent use of NSAIDs in rats. AB - AIM: To investigate the pharmacokinetic interactions between zalcitabine and nonsteroidal anti-inflammatory drugs (NSAIDs) in rats. METHODS: Zalcitabine was administered to rats via an iv injection (20 mg/kg) in the presence or absence of ketoprofen or naproxen (20 mg/kg), and the pharmacokinetic parameters were determined by using non-compartmental analysis. RESULTS: Compared with the control (zalcitabine alone), pretreatment with ketoprofen or naproxen 30 min prior to intravenous administration of zalcitabine significantly altered the pharmacokinetic profiles of zalcitabine in rats. Renal clearance of zalcitabine was reduced by approximately 3-4-fold in the presence of ketoprofen or naproxen. Consequently, systemic exposure (AUC) to zalcitabine in the rats pretreated with ketoprofen or naproxen was significantly greater than that for the control group given zalcitabine alone. The terminal plasma half-life of zalcitabine was also prolonged by 4-5-fold in the presence of ketoprofen or naproxen. CONCLUSION: The NSAIDs ketoprofen and naproxen effectively altered the pharmacokinetics of zalcitabine. Therefore, concomitant use of ketoprofen or naproxen in patients being treated with zalcitabine may necessitate close monitoring for potential drug interactions. PMID- 16364219 TI - Society for Reproductive Biology Founders' Lecture 2005. Control of metabolic cooperativity between oocytes and their companion granulosa cells by mouse oocytes. AB - Oocytes orchestrate the rate of follicular development and expression of genes in the surrounding granulosa cells. Oocytes are deficient in their ability to carry out some metabolic processes, such as glycolysis and amino acid uptake, and depend on the cooperation of granulosa cells to carry out these processes. In this dependency, the oocyte was previously considered a passive recipient of the nutritional support from granulosa cells. However, recent studies indicate an active role for the oocyte in controlling metabolic activity in granulosa cells. The ability of oocytes to control granulosa cell metabolism is achieved, at least in part, by regulating granulosa cell expression of genes encoding proteins involved in the metabolic processes. This review summarises current knowledge of intercellular communication between oocytes and granulosa cells from the perspective of oocyte control of gene expression in granulosa cells and metabolic cooperativity between the two cell types. The oocyte probably controls metabolism in granulosa cells to provide metabolites for its own development. In addition, we hypothesise that oocytes use their ability to regulate metabolic pathways in granulosa cells to orchestrate the rate of follicular development. PMID- 16364220 TI - Semen characteristics and sperm morphology in the Arabian leopard (Panthera pardus nimr) and how these vary with age and season. AB - The Arabian leopard is a critically endangered species. Since there are only an estimated 200 animals remaining in the wild, careful management of the captive population is necessary to minimise inbreeding. The objective of this study was to characterise sperm morphology and ejaculate quality in captive males. Semen was collected by electroejaculation from 8 adult captive male leopards (aged 2-16 years) during the summer and winter months, and semen parameters, including sperm morphology, were assessed. Two-year-old leopards showed lower total sperm counts per ejaculate than older animals and these counts declined at > 8 years. Ejaculates collected during the hot summer showed significantly lower sperm concentrations, total sperm counts, sperm motility and viability and percentage of spermatozoa showing normal morphology than ejaculates collected in the cooler winter. The results showed that the male leopard attains sexual maturity between 2 and 3 years of age and exhibits good semen quality until 8 years. Collection of semen for artificial breeding or banking would best be carried out in the cooler winter months. PMID- 16364221 TI - Validation of an experimental strategy for studying surface-exposed proteins involved in porcine sperm-oviduct contact interactions. AB - Previous experiments have shown that boar sperm survival in vitro is enhanced when co-incubated with a solubilised protein extract of porcine oviducal apical plasma membrane proteins. Here, we examine the hypothesis that the effects are mediated by direct oviduct-sperm contact and use in situ biotinylation of the oviducal epithelial surface to trace the surface-exposed biotinylated proteins through purification and solubilisation steps. We have also examined the effectiveness of mechanical scraping as a method of recovering oviducal epithelial proteins. We show that a subset of proteins originally exposed at the oviducal surface eventually bind to spermatozoa during incubation in vitro, but also show that a different protein subset is implicated if the sperm incubation is performed with proteins that had been biotinylated after (ex situ) extraction from the oviduct. Apical plasma membrane fractions biotinylated after purification contained many more biotinylated protein bands than preparations labelled before purification and multiple protein bands were eventually found to associate with spermatozoa. Although the evidence presented here supports the hypothesis that protein(s) anchored to the oviducal epithelium bind populations of spermatozoa directly and may have a role in the enhancement of sperm viability, it also shows that the choice of investigative technique exerts a major influence on experimental outcomes. PMID- 16364222 TI - Proteinase-activated receptors in ovine cervical function. AB - In sheep, inflammation not only functions in cervical dilation at parturition, but also plays an important part in the non-pregnant ewe cervix, as demonstrated by the high level of expression of interleukin (IL)-8 at oestrus. Ewes artificially induced to ovulate have significantly lower levels of IL-8 gene expression at oestrus compared with natural oestrus, indicating an inhibition of inflammation and function, offering an explanation for the low rates of conception in vaginally inseminated synchronised ewes. To identify potential pro inflammatory agents to combat the anti-inflammatory effects of hormonal synchronisation of oestrus, we have investigated the role of proteinase-activated receptor (PAR)-1 and PAR-2. To localise and measure the level of expression of these receptors, ovine-specific probes were derived for PAR-1 and PAR-2 and used for quantitative in situ hybridisation in the ovine cervix. Both PAR-1 and PAR-2 were expressed in the luminal epithelium of the cervix throughout the oestrous cycle, with expression being highest at oestrus. The gene expression of PAR-2 at oestrus was approximately 30% higher than that of PAR-1. Artificial synchronisation of oestrus by either an intravaginal progesterone sponge or prostaglandin F2+/- injections did not inhibit PAR-1 or PAR-2 expression at oestrus; rather, in the case of PAR-2, progesterone synchronisation increased it. Both synchronising procedures increased the expression of PAR-1 and PAR-2 during the luteal phase of the cycle. Therefore, agonists of PAR-1 and PAR-2 may be potentially useful pro-inflammatory agents countering the inhibition of inflammation by hormonal synchronisation. PMID- 16364223 TI - An efficient method of ovarian stimulation and in vitro embryo production from prepubertal lambs. AB - The production of embryos from prepubertal lambs is inefficient, partly resulting from the low developmental competence of prepubertal lamb oocytes, and partly because a high proportion of lambs fail to respond to hormone stimulation. The development of a hormone stimulation regimen that all lambs respond to would increase the efficiency of breeding from prepubertal animals. Using a hormone stimulation regimen consisting of oestradiol benzoate (50 microg), a norgestomet implant (1.5 mg), pregnant mare serum gonadotrophin (400 IU) and follicle stimulating hormone (130 mg) all lambs (n = 19) responded to hormone stimulation. Uterine and ovarian weight ranged from 2.8 to 7.2 g (11.8 +/- 0.7 g) and from 1.7 to 54.1 (12.5 +/- 2.9 g), respectively. The number of ovarian follicles and oocytes recovered ranged from 20.0 to 500.0 (118.2 +/- 29.2) and from 13.0 to 455.0 (82.0 +/- 24.2), respectively, and oocytes suitable for in vitro production were obtained from all 19 lambs. Uterine weight was related to both bodyweight and growth rate (P < 0.05), although ovarian weight and the number of ovarian follicles were not related to either bodyweight or growth rate. Oocyte cleavage varied between hormone-stimulated lambs (0.0-93.0%; P < 0.05), and 484/775 (62.2%) of the oocytes cultured cleaved. Oocytes from 17 of the 19 lambs (89.5%) developed to the blastocyst stage in vitro , and the proportion of zygotes forming a blastocyst (by Day 7) ranged from 0.0 to 66.7% for individual lambs. Overall, 33.9% of zygotes (n = 164) developed to the blastocyst stage, producing 8.6 +/- 2.8 blastocysts per lamb. PMID- 16364224 TI - Reproductive physiology and artificial insemination studies in wild and captive gerenuk ( Litocranius walleri walleri ). AB - Gerenuk antelope in North American zoos are descended from 28 founders imported from Kenya approximately 20 years ago. Intensive management is required to prevent inbreeding depression. Artificial insemination has potential for augmenting genetic management, but successful application requires a thorough understanding of species' reproductive norms. Semen collected from captive (n = 10) and wild (n = 6) gerenuk contained low numbers of morphologically normal spermatozoa (approximately 40%). Age, but not season, influenced (P < 0.05) the proportion of morphologically normal spermatozoa (mean +/- s.e.m., 12-17 months of age, 10.3 +/- 1.9%; 18-26 months of age, 34.4 +/- 6.2%; 3-6 years of age, 40.0 +/- 4.7%). Seasonality was investigated by analysing faecal testosterone and progesterone in males and females, respectively, by radioimmunoassays. Females cycled all year (ovarian cycle length, 18.7 +/- 0.9 days). Testosterone in males did not vary (P > 0.05) with time of year. Three females (3/9, 33%) became pregnant by insemination with 9.75-54.0 x 0(6) motile fresh or frozen sperm after oestrus synchronisation with two prostaglandin F(2alpha) injections, 12 days apart. One female inseminated with frozen-thawed sperm delivered a full-term stillborn calf after 213 days gestation. These results characterise gerenuk reproductive norms and indicate that artificial insemination may be a useful tool in the genetic management of gerenuk. PMID- 16364225 TI - Cobalt chloride, a hypoxia-mimicking agent, modulates redox status and functional parameters of cultured swine granulosa cells. AB - Hypoxia occurs physiologically during ovarian follicle growth; this deprivation represents a triggering stimulus for the production of vascular endothelial growth factor (VEGF) by proliferating granulosa cells, which are mostly responsible for the growth of the follicle. Moreover, the steroidogenic activity of these cells ensures a receptive environment for the implantation and development of the early embryo. The present paper reports the adaptive response of swine granulosa cells to cobalt chloride (CoCl(2), a chemical hypoxia mimicking agent. The effects of the treatment were evaluated on cell proliferation, steroidogenesis and VEGF production. In addition, because mithocondrial reactive oxygen species (ROS) are possibly involved in O(2) sensing, ROS levels and scavenging enzyme activity were investigated. In the present study, CoCl 2 had no effect on progesterone production, although it significantly reduced oestradiol synthesis. The addition of CoCl(2) to granulosa cell culture stimulated VEGF production and the generation of hydrogen peroxide. Chemical hypoxia had different effects on scavenger enzyme activities: the activity of superoxide dismutase was enhanced, that of peroxidase reduced and catalase activity was unaffected. The net result is a 'pro-oxidant' state, which appears to be possibly involved in the stimulation of VEGF production, thus inducing follicular angiogenesis. PMID- 16364227 TI - Studies on the cryopreservation of common wombat (Vombatus ursinus) spermatozoa. AB - In an attempt to develop a gamete-recovery protocol for the northern hairy nosed wombat ( Lasiorhinus krefftii ), spermatozoa were removed from the cauda epididymides of four common wombats ( Vombatus ursinus ) and cryopreserved following a variety of prefreeze storage conditions. Spermatozoa stored for 72 h at 4 degrees C within the testicle before cryopreservation tolerated the freeze thaw procedure remarkably well, resulting in a higher post-thaw viability (% motile P < 0.01; rate of movement P < 0.01; % live P < 0.01) than sperm recovered on the day of post-mortem, stored in a test tube for 72 h at 4 degrees C and then frozen. The effect of post-thaw dilution with Tris citrate fructose (TCF) diluent on the survival of epididymal common wombat spermatozoa was also investigated. Motility (P < 0.05), rate of sperm movement (P < 0.01) and the percentage of live spermatozoa (P < 0.05) were all significantly greater when spermatozoa were thawed and diluted immediately in TCF than when thawed without dilution. The present study also reports, for the first time, a successful pellet method of freezing wombat spermatozoa on dry ice; volumes of 0.25 and 0.5 mL resulted in higher post-thaw survival compared with 0.1-mL pellets. PMID- 16364226 TI - Corpus luteum life span and pituitary oestrogen and progesterone receptors in cyclic and gonadotrophin-releasing hormone-treated anoestrous ewes. AB - The present study investigated the pituitary oestrogen (ER) and progesterone (PR) receptor concentrations in ewes during the oestrous cycle in the breeding season (n = 19), and in anoestrous ewes treated with gonadotrophin-releasing hormone (GnRH) (n = 11) and anoestrous ewes treated with progesterone + GnRH (n = 11). The pituitary ER and PR concentrations at the expected time of ovulation and in the early and late luteal phases were measured by binding assay. The pattern of pituitary ER and PR concentrations in the progesterone + GnRH-treated ewes resembled the pattern found during the normal oestrous cycle, with ER and PR concentrations decreasing from the time of ovulation to the early luteal phase. In contrast, in ewes treated with GnRH alone, ER and PR concentrations increased in the early luteal phase, which may increase the inhibitory effects of steroid hormones on luteinising hormone secretion, ultimately leading to the development of subnormal luteal phases. PMID- 16364228 TI - Characterisation of the progesterone receptor on canine spermatozoa. AB - The present study was conducted to characterise and localise the progesterone receptor (PR) on canine spermatozoa. Using a progesterone-bovine serum albumin fluorescein isothiocyanate conjugate (PBF) and different monoclonal antibodies (C262 and NCL-PGR against the steroid binding domain and N-terminus of intracellular PR, respectively, and h151 against the hinge domain of the intracellular oestrogen receptor), the PR was identified on the plasma membrane over the acrosomal region. Two proteins (54 kDa and 65 kDa) were detected by recognition of the three monoclonal antibodies using Western blotting. PBF labelling was observed in the majority of cauda epididymal spermatozoa (63 +/- 4%), but this labelling was markedly reduced (33 +/- 17%) after the addition of canine seminal plasma. Over a 7-h capacitation, the proportion of ejaculated spermatozoa exhibiting PBF labelling (indicating the presence of the PR) increased from 18 +/- 10% (onset) to 59 +/- 7% by 5 h, where it plateaued. Progesterone (P 4 ) induced the acrosome reaction (AR) in a dose-dependent manner (0, 0.1, 1 and 10 ug/mL P 4 corresponding to 10 +/- 5%, 16 +/- 9%, 23 +/- 7% and 30 +/- 7%). Pre-treatment of capacitated spermatozoa with canine seminal plasma reduced the incidence of the P 4 -induced AR (12 +/- 5%). In addition, treatment with the monoclonal antibodies significantly reduced the incidence of the P 4 induced AR (10 microg/mL) in capacitated ejaculated spermatozoa from 19 +/- 6% to 11 +/- 4% (h151, 1 : 10) and 12 +/- 6% (C262, 1 : 10), respectively. A typical Scatchard plot revealed one binding with high affinity and low capacity, and another binding with low affinity and high capacity, suggesting at least two different characteristic PR. Taken together, these results demonstrate that P 4 induced the AR in a dose-dependent manner via functional transmembranal receptors in the acrosomal region of the canine sperm plasma membrane. The characteristics of this membrane receptor seem similar to those of other mammalian spermatozoa, and it shows structural homology to the intracellular PR. PMID- 16364229 TI - Glucocorticoids and serotonin alter glucocorticoid receptor mRNA levels in fetal guinea-pig hippocampal neurons, in vitro. AB - The hypothalamic-pituitary-adrenal (HPA) axis is susceptible to programming during fetal life. Such programming occurs, at least partially, at the level of the hippocampus. The hippocampus plays a central role in regulation of the HPA axis and release of endogenous glucocorticoids, via mediation of glucocorticoid negative feedback. Fetal exposure to synthetic glucocorticoids can permanently alter glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) levels within the hippocampus, and serotonin is thought to be involved in this process. In the present study, we hypothesised that dexamethasone, cortisol and serotonin exposure would modify GR mRNA expression within fetal guinea-pig hippocampal cultures. Cultures were derived from 40-day-old guinea-pig fetuses, and were exposed to 0, 1, 10 and 100 nM dexamethasone, cortisol or serotonin for 4 days. Expression of GR and MR mRNA was examined by in situ hybridisation followed by high-resolution silver emulsion autoradiography. Four-day exposure to dexamethasone (P < 0.05; 100 nM) or cortisol (P = 0.08; 100 nM) downregulated the expression of GR mRNA within neurons. There was no change in the expression of MR mRNA levels following cortisol treatment. Exposure to serotonin (100 nM) significantly increased GR mRNA levels in hippocampal neurons. We conclude that synthetic and endogenous glucocorticoids, as well as serotonin, can influence GR expression during hippocampal development and in this way may act to permanently programme HPA function. PMID- 16364230 TI - [Medicoeconomic evaluation of the treatment of morbid obesity by Swedish adjustable gastric banding (SAGB)]. AB - INTRODUCTION: Aim of the present study was to analyze the medicoeconomic impact of each treatment for obesity in France, taking into account morbid obesity associated comorbidities from a health insurance perspective and to calculate the cost-effectiveness ratio of SAGB compared to the non-surgical treatment for various kinds of patients, as well as the budget impact on a given cohort of patients. METHODS: The model studied surgery-eligible patients and compared the effectiveness of the SAGB treatment to the conventional treatment. The follow-up extended from 1 to 5 years. Costs analyzed were treatment-related direct medical costs, as well as potential comorbidities costs. The effectiveness is determined according to the BMI loss and its maintenance over time, together with the level of improved quality of life. RESULTS: An evaluation on a series of 1,000 patients shows that the treatment with SAGB is dominant (less expensive, more effective in terms of loss of BMI and its duration) compared to the conventional treatment for patients suffering from type II diabetes or obstructive sleep apnea. The same evaluation in terms of QALYs shows that the treatment with SAGB is dominant compared to the conventional treatment for patients with a BMI >or= 35 kg/m(2) and a type II diabetes, as well as for patients whose BMI >or= 40 kg/m(2) (with or without type II diabetes mellitus). PMID- 16364231 TI - Alarm timing, trust and driver expectation for forward collision warning systems. AB - In order to improve road safety, automobile manufacturers are now developing Forward Collision Warning Systems (FCWS). However, there has been insufficient consideration of how drivers may respond to FCWS. This driving simulator study focused on alarm timing and its impact on driver response to alarm. The experimental investigation considered driver perception of alarm timings and its influence on trust at three driving speeds (40, 60 and 70 mile/h) and two time headways (1.7 and 2.2 s). The results showed that alarm effectiveness varied in response to driving conditions. Alarm promptness had a greater influence on ratings of trust than improvements in braking performance enabled by the alarm system. Moreover, alarms which were presented after braking actions had been initiated were viewed as late alarms. It is concluded that drivers typically expect alarms to be presented before they initiate braking actions and when this does not happen driver trust in the system is substantially decreased. PMID- 16364232 TI - Mechanism of Gly-Pro-pNA cleavage catalyzed by dipeptidyl peptidase-IV and its inhibition by saxagliptin (BMS-477118). AB - Dipeptidyl peptidase-IV (DPP-IV) is a serine protease with a signature Asp-His Ser motif at the active site. Our pH data suggest that Gly-Pro-pNA cleavage catalyzed by DPP-IV is facilitated by an ionization of a residue with a pK of 7.2 +/- 0.1. By analogy to other serine proteases this pK is suggestive of His-Asp assisted Ser addition to the P1 carbonyl carbon of the substrate to form a tetrahedral intermediate. Solvent kinetic isotope effect studies yielded a D2Okcat/Km=2.9+/-0.2 and a D2Okcat=1.7+/-0.2 suggesting that kinetically significant proton transfers contribute to rate limitation during acyl intermediate formation (leaving group release) and hydrolysis. A "burst" of product release during pre steady-state Gly-Pro-pNA cleavage indicated rate limitation in the deacylation half-reaction. Nevertheless, the amplitude of the burst exceeded the enzyme concentration significantly (approximately 15-fold), which is consistent with a branching deacylation step. All of these data allowed us to better understand DPP-IV inhibition by saxagliptin (BMS-477118). We propose a two-step inhibition mechanism wherein an initial encounter complex is followed by covalent intermediate formation. Final inhibitory complex assembly (kon) depends upon the ionization of an enzyme residue with a pK of 6.2 +/- 0.1, and we assigned it to the catalytic His-Asp pair which enhances Ser nucleophilicity for covalent addition. An ionization with a pK of 7.9 +/- 0.2 likely reflects the P2 terminal amine of the inhibitor hydrogen bonding to Glu205/Glu206 in the enzyme active site. The formation of the covalent enzyme-inhibitor complex was reversible and dissociated with a koff of (5.5 +/- 0.4) x 10(-5) s(-1), thus yielding a Ki* (as koff/kon) of 0.35 nM, which is in good agreement with the value of 0.6 nM obtained from steady-state inhibition studies. Proton NMR spectra of DPP-IV showed a downfield resonance at 16.1 ppm. Two additional peaks in the 1H NMR spectra at 17.4 and 14.1 ppm were observed upon mixing the enzyme with saxagliptin. Fractionation factors (phi) of 0.6 and 0.5 for the 17.4 and 14.1 ppm peaks, respectively, are suggestive of short strong hydrogen bonds in the enzyme inhibitor complex. PMID- 16364233 TI - Probing interactions between the coagulants thrombin, Factor XIII, and fibrin(ogen). AB - Thrombin cleaves fibrinopeptides A and B from fibrinogen leading to the formation of a fibrin network that is later covalently crosslinked by Factor XIII (FXIII). Thrombin helps activate FXIII by catalyzing hydrolysis of the FXIII activation peptides (AP). In the current work, the role of exosites in the ternary thrombin FXIII-fibrin(ogen) complex was further explored. Hydrolysis studies indicate that thrombin predominantly utilizes its active site region to bind extended Factor XIII AP (FXIII AP 33-64 and 28-56) leaving the anion-binding exosites for fibrin(ogen) binding. The presence of fibrin-I leads to improvements in the K(m) for hydrolysis of FXIII AP (28-41), whereas peptides based on the cardioprotective FXIII V34L sequence exhibit less reliance on this cofactor. Surface plasmon resonance measurements reveal that d-Phe-Pro-Arg chloromethylketone-thrombin binds to fibrinogen faster than to FXIII a(2) and dissociates from fibrinogen more slowly than from FXIII a(2). This system of thrombin exosite interactions with differing affinities promotes efficient clot formation. PMID- 16364234 TI - Fucoidan a sulfated polysaccharide from brown algae is a potent modulator of connective tissue proteolysis. AB - Fucoidans are sulfated fucosylated polymers from brown algae cell wall that exhibit some heparin/heparan sulfate properties. We previously demonstrated that these polysaccharides were able in vitro to stimulate dermal fibroblast proliferation and extracellular matrix deposition. Here, we investigated the action of a 16kDa fucoidan fraction on parameters involved in connective tissue breakdown. This fucoidan is able to inhibit gelatinase A secretion and stromelysin 1 induction by interleukin-1beta on dermal fibroblasts in culture. Furthermore, we observed that fucoidan increases the rate of association of MMPs with their specific inhibitors namely TIMPs. Using tissue sections of human skin in ex vivo experiments, we evidenced that this polysaccharide was able to minimize human leukocyte elastase activity resulting in the protection of human skin elastic fiber network against the enzymatic proteolysis due to this serine proteinase. These results suggested that fucoidan could be used for treating some inflammatory pathologies in which uncontrolled extracellular matrix degradation takes place. PMID- 16364235 TI - Analysis of photosynthetic complexes from a cyanobacterial ycf37 mutant. AB - The Ycf37 protein has been suggested to be involved in the biogenesis and/or stability of the cyanobacterial photosystem I (PSI). With Ycf37 specific antibodies, we analyzed the localization of Ycf37 within the thylakoid membranes of the cyanobacterium Synechocystis sp. PCC 6803. Inspection of a sucrose gradient profile indicated that small amounts of Ycf37 co-fractionated with monomeric photosynthetic complexes, but not with trimeric PSI. Isolating 3xFLAG epitope-tagged Ycf37 by affinity-tag purification rendered several PSI subunits that specifically co-precipitated with this protein. Blue-native PAGE newly revealed two monomeric PSI complexes (PSI and PSI*) in wild-type thylakoids. The lower amount of PsaK present in PSI* may explain its higher electrophoretic mobility. PSI* was more prominent in high-light grown cells and interestingly proved absent in the Deltaycf37 mutant. PSI* appeared again when the mutant was complemented in trans with the wild-type ycf37 gene. In the Deltaycf37 mutant the amount of trimeric PSI complexes was reduced to about 70% of the wild-type level with no significant changes in photochemical activity and subunit composition of the remaining photosystems. Our results indicate that Ycf37 plays a specific role in the preservation of PSI* and the biogenesis of PSI trimers. PMID- 16364236 TI - Phenothiazine maleates stimulate MRP1 transport activity in human erythrocytes. AB - The expression of multidrug resistance-associated protein (MRP1) results in ATP dependent reduction of drugs' concentration in cancer cells, i.e., multidrug resistance (MDR). Since the majority of projects are concentrated on the search of the new MDR modulators, there are very few reports on drug-induced stimulation of MDR transporters activity. In the present work, by means of functional fluorescence assay we have shown that MRP1-mediated efflux of 2',7'-bis-(3 carboxypropyl)-5-(and-6)-carboxyfluorescein (BCPCF) out of human erythrocytes is stimulated by phenothiazine maleates that have been already identified as P glycoprotein inhibitors. Phenothiazine maleates-induced stimulation of ATP dependent uptake of 2',7'-bis-(3-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) into inside-out membrane vesicles prepared from erythrocyte membranes has been also demonstrated. Moreover, it was shown that phenothiazine maleates exerted stimulating effect on ATPase activity measured in erythrocyte membranes. To our best knowledge, this report is the first one demonstrating that compounds able to inhibit transport activity of P-glycoprotein can stimulate MRP1 transporter. We conclude that phenothiazine maleates probably exert their stimulatory effect on MRP1 by direct interaction with the protein at the site different from the substrate binding site. PMID- 16364237 TI - Sex differences in response to a panicogenic challenge procedure: an experimental evaluation of panic vulnerability in a non-clinical sample. AB - Epidemiological studies show that women are twice as likely as men to develop panic disorder (PD) during their lifetimes. Data from retrospective studies also suggest that women are more distressed by panic and other negative emotional states than men, and that this tendency may precede the development of PD. The present prospective study sought to expand this work by evaluating sex differences in the predisposition to panic in individuals without PD or other forms of psychopathology. Participants (N=96; 52 women) were exposed to 12 panicogenic inhalations of 20% CO2-enriched air while physiological (e.g., heart rate, electrodermal response, Frontalis EMG) and self-report (e.g., subject distress, fear, frequency and severity of DSM-IV panic symptoms) response domains were monitored. As expected, magnitude of autonomic responding failed to reliably discriminate between women and men. Yet, women reported more fear and panic immediately following the challenge procedure relative to men, and this sex difference persisted when assessed again 30 min post-challenge. Implications of these results are discussed broadly in the context of biopsychosocial accounts of sex differences in the clinical presentation of PD. PMID- 16364238 TI - Formation of albumin dimers induced by exposure to peroxides in human plasma: a possible biomarker for oxidative stress. AB - Human serum albumin (HSA) has one free thiol residue at Cys-34 that is likely oxidized by various reactive oxygen species (ROS). We attempted to identify the oxidation product of Cys-34 of HSA following exposure of plasma to ROS. Oxidation induced by tert-butyl hydroperoxide (t-BuOOH) of this free cysteine residue in HSA was observed in detail. Analysis of oxidized albumin in a partially purified fraction obtained by affinity column chromatography clearly revealed the formation of albumin disulfide dimers following t-BuOOH exposure. Albumin disulfide dimer formation was observed in normal plasma following treatment with various peroxides, as well as in untreated plasma from patients on hemodialysis using SDS-PAGE and Western blot analysis. The present results indicate that albumin dimers are oxidative products derived from peroxides, and that their presence in plasma might be a marker of oxidative stress as secondary metabolites of peroxidation. PMID- 16364239 TI - Interactions among the components of the interleukin-10 receptor complex. AB - We used fluorescence resonance energy transfer previously to show that the interferon-gamma (IFN-gamma) receptor complex is a preformed entity mediated by constitutive interactions between the IFN-gammaR2 and IFN-gammaR1 chains, and that this preassembled entity changes its structure after the treatment of cells with IFN-gamma. We applied this technique to determine the structure of the interleukin-10 (IL-10) receptor complex and whether it undergoes a similar conformational change after treatment of cells with IL-10. We report that, like the IFN-gamma receptor complex, the IL-10 receptor complex is preassembled: constitutive but weaker interactions occur between the IL-10R1 and IL-10R2 chains, and between two IL-10R2 chains. The IL-10 receptor complex undergoes no major conformational changes when cells are treated with cellular or Epstein-Barr viral IL-10. Receptor complex preassembly may be an inherent feature of Class 2 cytokine receptor complexes. PMID- 16364240 TI - Role of N-glycan-dependent quality control in the cell-surface expression of the AT1 receptor. AB - Most G protein-coupled receptors (GPCRs) are N-glycosylated proteins but the role of this post-translational modification in GPCR biosynthesis has not been extensively studied. We previously showed that the non-glycosylated AT(1) receptor is inefficiently expressed at the cell surface. In this study, we addressed whether AT(1) interacts with elements of the ER-based quality control processes. Interestingly, non-glycosylated AT(1) receptors associated with the molecular chaperones calnexin and HSP70, suggesting the importance of protein based interactions between these partners. We also demonstrate that ER mannosidase I participates in the acquisition of mature glycoforms and in the targeting of the AT(1) receptor to the membrane. Taken together, these results indicate that decreased cell-surface expression of the non-glycosylated receptor cannot be attributed to diminished interactions with molecular chaperones and that mannose trimming of the wild-type AT(1) receptor by ER mannosidase I plays a critical role in its cell-surface expression. PMID- 16364241 TI - Ran binding protein 9 interacts with Raf kinase but does not contribute to downstream ERK1/2 activation in skeletal myoblasts. AB - Raf kinase is the upstream activator of MEK1/2 leading to phosphorylation and activation of ERK1/2. Sustained activation of Raf represses skeletal muscle specific reporter gene transcription and formation of multinucleated myofibers. Inhibition of myogenesis by activated Raf involves downstream ERK1/2 as well as undefined mediators. To identify Raf-interacting proteins that may influence repression of muscle formation, a yeast two-hybrid screen was performed using a MEK1-binding defective Raf (RafBXB-T481A) as bait. Twenty cDNAs coding for Raf interacting proteins were identified including Ran binding protein 9 (RanBP9), a protein previously reported to interact with receptor tyrosine kinases. Forced expression of RanBP9 in myogenic cells did not alter myogenesis. Co-expression of RanBP9 with constitutively active RafBXB, but not RafBXB-T481A, synergistically inhibited MyoD-directed muscle reporter gene transcription. Knockdown of RanBP9 expression did not restore the differentiation program to Raf-expressing myoblasts. Thus, RanBP9 physically associates with Raf but does not substantially contribute to the inhibitory actions of the kinase. PMID- 16364242 TI - Curcumin induces growth-arrest and apoptosis in association with the inhibition of constitutively active JAK-STAT pathway in T cell leukemia. AB - Adult T cell leukemia is an aggressive and frequently fatal malignancy that expressess constitutively activated growth-signaling pathways in association with deregulated growth and resistance to apoptosis. Curcumin (diferuloylmethane) is a naturally occurring yellow pigment, isolated from the rhizomes of the plant Curcuma longa that has traditionally been used in the treatment of injury and inflammation. But the effect and mechanism of action of curcumin on T cell leukemia is not known. To investigate the antitumor activity of curcumin in T cell leukemia, we examined its effect on constitutive phosphorylation of JAK and STAT proteins, proliferation, and apoptosis in HTLV-I-transformed T cell lines. HTLV-I-transformed T cell leukemia lines, MT-2, HuT-102, and SLB-1, express constitutively phosphorylated JAK3, TYK2, STAT3, and STAT5 signaling proteins. In vitro treatment with curcumin induced a dose-dependent decrease in JAK and STAT phosphorylation resulting in the induction of growth-arrest and apoptosis in T cell leukemia. The induction of growth-arrest and apoptosis in association with the blockade of constitutively active JAK-STAT pathway suggests this be a mechanism by which curcumin induces antitumor activity in T cell leukemia. PMID- 16364243 TI - Heritable alteration in DNA methylation pattern occurred specifically at mobile elements in rice plants following hydrostatic pressurization. AB - Intrinsic DNA methylation pattern is an integral component of the epigenetic network in many eukaryotes. Exploring the extent to which DNA methylation patterns can be altered under a specific condition is important for elucidating the biological functions of this epigenetic modification. This is of added significance in plants wherein the newly acquired methylation patterns can be inherited through organismal generations. We report here that DNA methylation patterns of mobile elements but not of cellular genes were specifically altered in rice plants following hydrostatic pressurization. This was evidenced by methylation-sensitive gel-blot analysis, which showed that 10 out of 10 studied low-copy transposons and retrotransposons manifested methylation alteration in at least one of the 8 randomly chosen pressure-treated plants, whereas none of the 16 studied low-copy cellular genes showed any change. Both gel-blotting and genome-wide fingerprinting indicated that the methylation alteration in mobile elements was not accompanied by a general genetic instability. Progeny analysis indicated retention of the altered methylation patterns in most progeny plants, underscoring early occurrence of the alterations, and their faithful epigenetic inheritance. PMID- 16364244 TI - Leber's hereditary optic neuropathy is associated with the mitochondrial ND4 G11696A mutation in five Chinese families. AB - We report here the clinical, genetic, and molecular characterization of five Chinese families with Leber's hereditary optic neuropathy (LHON). Clinical and genetic evaluations revealed the variable severity and age-of-onset in visual impairment in these families. Strikingly, there were extremely low penetrances of visual impairment in these Chinese families. Sequence analysis of the complete mitochondrial genomes in these pedigrees showed the distinct sets of mtDNA polymorphism, in addition to the identical ND4 G11696A mutation associated with LHON. Indeed, this mutation is present in homoplasmy only in the maternal lineage of those pedigrees but not other members of these families. In fact, the occurrence of the G11696A mutation in these several genetically unrelated subjects affected by visual impairment strongly indicates that this mutation is involved in the pathogenesis of visual impairment. Furthermore, the N405D in the ND5 and G5820A in the tRNA(Cys), showing high evolutional conservation, may contribute to the phenotypic expression of G11696A mutation in the WZ10 pedigree. However, there was the absence of functionally significant mtDNA mutations in other four Chinese pedigrees carrying the G11696A mutation. Therefore, nuclear modifier gene(s) or environmental factor(s) may play a role in the phenotypic expression of the LHON-associated G11696A mutation in these Chinese pedigrees. PMID- 16364245 TI - A test of Chargaff's second rule. AB - In 1968, Chargaff and his colleagues discovered a rule in Bacillus subtilis: in single stranded DNA, A=T and C=G. This rule has since been confirmed many times in other bacterial and eukaryotic genomes. To the best of our knowledge, this rule has not been tested before in either single stranded DNA or RNA genomes. Over 3400 genomic sequences were examined here and included for the first time both double and single stranded DNA and RNA genomes. We found that: (1) with the exception of the organellar DNA, this parity rule holds for all types of double stranded DNA genomes and (2) that this rule fails to hold for other types of genomes. The parity rule appears to be a selective force on genome evolution and codon use. PMID- 16364247 TI - Improving effect of ethyl eicosapentanoate on statin-induced rhabdomyolysis in Eisai hyperbilirubinemic rats. AB - The effect of ethyl eicosapentanoate (EPA-E) on statin-induced rhabdomyolysis was investigated by co-administration of EPA-E and pravastatin (PV), as a typical statin, to Eisai hyperbilirubinemic rats (EHBR). It was confirmed that the plasma PV concentration was not affected by simultaneous administration of EPA-E, and there was no cumulative increase of PV during prolonged co-administration of EPA E and PV. Muscular degeneration was prominent (incidence 5/5; average grade 3.5 (range 2-4)) in EHBR treated with PV alone at 200 mg/kg/day for 14 days, but co administration of EPA-E at doses of 100, 300, and 1000 mg/kg/day decreased the average grades to 1.4 (range 0.3-3.0), 0.5 (0.2-1.0), and 0.6 (0.0-1.7), respectively. Creatine phosphokinase (CPK) and myoglobin levels in plasma were well correlated with the grade of skeletal muscle degeneration. Thus, EPA-E appears to reduce the severity of statin-induced rhabdomyolysis. PMID- 16364246 TI - Insulin and oleic acid increase PPARgamma2 expression in cultured mouse hepatocytes. AB - Hepatic PPARgamma expression is increased in several animal models of diabetes and obesity, and liver-specific overexpression of PPARgamma induces liver steatosis. The aim of this study was to investigate the regulation of PPARgamma expression in primary mouse hepatocytes. PPARgamma2, but not PPARgamma1, was up regulated by insulin and to a lesser extent by oleic acid. Insulin increased transcription of the PPARgamma2 gene via phosphatidylinositol 3-kinase activation. The PPARgamma agonist, rosiglitazone, increased PPARgamma2 expression, but not PPARgamma1, only in the presence of insulin. Also aP2 mRNA expression was induced by rosiglitazone to a higher degree in the presence of insulin, while acyl-CoA oxidase was increased independently of insulin. In summary, PPARgamma2 is increased in hepatocytes by oleic acid and insulin. These results may help to understand the regulation of PPARgamma expression in liver, which possibly plays a role in the development of liver steatosis. PMID- 16364248 TI - Enhanced transformation and chemosensitivity of NIH3T3 cells transduced with hepatoma up-regulated protein. AB - Hepatoma up-regulated protein (HURP) is a recently identified novel cell-cycle regulated gene. The HURP gene is overexpressed in human hepatocellular carcinoma and transitional cell carcinoma. The cellular function of HURP is not fully understood. In this study, the NIH3T3 cells transduced with the exogenous HURP gene manifested the general characteristics of tumor cells, which had higher growth rate in low-serum media and advanced ability of colony formation on agarose-based plates. Transduced HURP was capable of specifically enhancing the chemosensitivity of deoxycytosine analogs, such as gemcitabine, ARA-C, and 5-AZA CdR, but neither had an effect on the response of DNA intercalating agents, such as cisplatin, carboplatin, and doxorubicin, nor on the response of microtubule stabilizers, such as paclitaxel, docetaxel, and vinblastine. These results indicate that the HURP gene might be a potential oncogenic gene and capable of enhancing the chemosensitivity of deoxycytosine analogs in NIH3T3 cells. PMID- 16364249 TI - TP53 and P21 polymorphisms: response to cisplatinum/paclitaxel-based chemotherapy in ovarian cancer. AB - Ovarian cancer (OC) is the most lethal gynaecologic cancer and its standard treatment consists of platinum-based chemotherapy after cytoreductive surgery. The p53 protein plays a critical role on different cellular processes in response to DNA damage and it is responsible for transcriptional induction of the P21 gene. We have analysed 114 blood samples in order to investigate the effect of the TP53 codon 72 and the P21 3'UTR polymorphisms in response to cisplatinum/paclitaxel chemotherapy for OC treatment. The genotypes of the TP53 codon 72 and P21 3'UTR polymorphism were identified using AS-PCR and PCR-RFLP, respectively. Our results indicate that the TP53 P allele is associated with a worse prognosis (P=0.011) while P21 polymorphism genotypes did not reveal any statistically significant result (P>0.05). Furthermore, simultaneous carriers of the TP53 AA genotype and the P21 CC genotype demonstrate a longer progression free interval (P=0.020). This study suggests that the characterisation of a genetic profile can contribute to the definition of a better chemotherapy treatment. PMID- 16364251 TI - Transcriptional silencing of EphB6 receptor tyrosine kinase in invasive breast carcinoma cells and detection of methylated promoter by methylation specific PCR. AB - The receptor tyrosine kinase EphB6 is expressed at reasonable levels in normal breast cells. It shows decreased abundance in non-invasive breast carcinoma cells and is transcriptionally silenced in invasive breast carcinoma cells. We have characterized EphB6 promoter and correlated the expression of EphB6 transcript to differential methylation of the promoter region. The demethylation of promoter sequence in vivo by growth in media containing 5-aza-2'-deoxycytidine restores the expression of EphB6 to normal levels in breast carcinoma cells, and the ability of the promoter to initiate transcription of a reporter gene is lost after methylation of the promoter sequence. The promoter region has binding sites for various factors such as SP1 and p300. The specific methylation of CpG dinucleotides has allowed us to design primers that can selectively amplify the methylated promoter and thus facilitate identification of normal, non-invasive, and invasive breast cells. The potential significance of EphB6 to serve as a diagnostic and prognostic indicator is discussed. PMID- 16364250 TI - In vivo delivery of small interfering RNA targeting brain capillary endothelial cells. AB - Brain capillary endothelial cells (BCECs) play an important role in blood-brain barrier (BBB) functions and pathophysiologic mechanisms in brain ischemia and inflammation. We try to suppress gene expression in BCECs by intravenous application of small interfering RNA (siRNA). After injection of large dose siRNA with hydrodynamic technique to mouse, suppression of endogenous protein and the BBB function of BCECs was investigated. The brain-to-blood transport function of organic anion transporter 3 (OAT3) that expressed in BCECs was evaluated by Brain Efflux Index method in mouse. The siRNA could be delivered to BCECs and efficiently inhibited endogenously expressed protein of BCECs. The suppression effect of siRNA to OAT3 is enough to reduce the brain-to-blood transport of OAT3 substrate, benzylpenicillin at BBB. The in vivo siRNA-silencing method with hydrodynamic technique may be useful for the study of BBB function and gene therapy targeting BCECs. PMID- 16364252 TI - Volume-sensitive outwardly rectifying chloride channels are involved in oxidative stress-induced apoptosis of mesangial cells. AB - Volume-sensitive outwardly rectifying (VSOR) Cl- channels have been electrophysiologically identified in human and mouse mesangial cells, but the functional role of VSOR Cl- channels in mesangial cell apoptosis is not clear. The aim of the present study was to demonstrate the role of VSOR Cl- channels in oxidative stress-induced mesangial cell apoptosis. H2O2-induced Cl- currents showed phenotypic properties of VSOR Cl- channels, including outward rectification, voltage-dependent inactivation at more positive potentials, sensitivity to hyperosmolarity, and inhibition by VSOR Cl- channel blockers. Moreover, blockage of VSOR Cl- channels by DIDS (100 microM), NPPB (10 microM) or niflumic acid (10 microM) rescued mesangial cell apoptosis induced by H2O2. Treatment with 150 microM H2O2 for 2h resulted in significant reduction of cell volume, in contrast, nuclear condensation and/or fragmentation were not observed and the caspase-3 activity was also not increased. The early-phase alterations in cell volume were markedly abolished by pretreatment with VSOR Cl- channel blockers. We conclude that VSOR Cl- channels are involved in H2O2-induced apoptosis in cultured mesangial cells and its mechanism is associated with apoptotic volume decrease processes. PMID- 16364254 TI - Inhibition of mTOR signaling with rapamycin attenuates renal hypertrophy in the early diabetic mice. AB - Early diabetic nephropathy is characterized by renal hypertrophy that is mainly due to proximal tubular hypertrophy. Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase, and its signaling has been reported to regulate protein synthesis and cellular growth, specifically, hypertrophy. Therefore, we examined the effect of mTOR signaling on diabetic renal hypertrophy by using the specific inhibitor for mTOR, rapamycin. Ten days after streptozotocin-induced diabetes, mice showed kidney hypertrophy with increases in the phosphorylation of p70S6kinase and the expression of cyclin kinase inhibitors, p21(Cip1) and p27(Kip1), in the kidneys. The intraperitoneal injection of rapamycin (2 mg/kg/day) markedly attenuated the enhanced phosphorylation of p70S6kinase, the increment of cyclin-dependent kinase inhibitors, and renal enlargement without any changes of clinical parameters, including blood glucose, blood pressure, and food intake. Overexpression of a constitutive active form of p70S6kinase resulted in increased cell size of cultured mouse proximal tubule cells; thus, activation of p70S6kinase causes hypertrophy of proximal tubular cells. Our findings suggest that activation of mTOR signaling causes renal hypertrophy at the early stage of diabetes. PMID- 16364253 TI - AMPK activation increases fatty acid oxidation in skeletal muscle by activating PPARalpha and PGC-1. AB - AMP-activated protein kinase (AMPK) activation increases fatty acid oxidation in skeletal muscle by decreasing malonyl CoA concentrations. However, this may not explain the long-term effects of AMPK activation. Here we show that AMPK activation by 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) increases mRNA expression of PPARalpha target genes and PGC-1 in cultured muscle cells and mouse skeletal muscle, and that inhibition of PPARalpha and PGC-1 by siRNAs prevents AICAR-stimulated increase in fatty acid oxidation. These data suggest that a novel transcriptional regulatory mechanism involving PPARalpha and PGC-1 exists that is responsible for long-term stimulation of fatty acid oxidation in skeletal muscle by AICAR. PMID- 16364256 TI - Selection of ovine PrP high-producer subclones from a transfected epithelial cell line. AB - The hallmarks of prion diseases are the conversion of the normal prion into an abnormal protease resistant isoform and its brain accumulation. Purification of the native abnormal prion isoform for biochemical and biophysical studies has been hampered by poor recovery from brain tissue. An epithelial cell transfected with the ovine VRQ allele prion, called Rov9, has been used to select prion high producer cells by flow cytometry. The representative clone 4 described here produced 6.2 microg of cellular prion protein per mg of total protein extract, representing 8- to 10-fold the amount produced by the Rov9 parental cells. After exposure to the scrapie agent (PG128/98), clone 4 produced 2.6 microg of abnormal isoform per mg of total protein. When infected clone 4 cell cultures were treated with tunicamycin, 80% of the abnormal isoform was deglycosylated. The infectivity of the prions produced in clone 4 cultures was confirmed in a mouse bioassay. Such high-producer clones represent new tools for producing large amounts of glycosylated and/or non-glycosylated PrP(Sc) and for a powerful screening of clinical samples' infectivity. PMID- 16364255 TI - Cytoplasmic sequestration of cyclin D1 associated with cell cycle withdrawal of neuroblastoma cells. AB - The regulation of D-type cyclin-dependent kinase activity is critical for neuronal differentiation and apoptosis. We recently showed that cyclin D1 is sequestered in the cytoplasm and that its nuclear localization induces apoptosis in postmitotic primary neurons. Here, we further investigated the role of the subcellular localization of cyclin D1 in cell cycle withdrawal during the differentiation of N1E-115 neuroblastoma cells. We show that cyclin D1 became predominantly cytoplasmic after differentiation. Targeting cyclin D1 expression to the nucleus induced phosphorylation of Rb and cdk2 kinase activity. Furthermore, cyclin D1 nuclear localization promoted differentiated N1E-115 cells to reenter the cell cycle, a process that was inhibited by p16(INK4a), a specific inhibitor of D-type cyclin activity. These results indicate that cytoplasmic sequestration of cyclin D1 plays a role in neuronal cell cycle withdrawal, and suggests that the abrogation of machinery involved in monitoring aberrant nuclear cyclin D1 activity contributes to neuronal tumorigenesis. PMID- 16364257 TI - Endothelins in the cat petrosal ganglion and carotid body: effects and immunolocalization. AB - In response to hypoxia, chemoreceptor cells of the carotid body (CB) release transmitters, which acting on the petrosal ganglion (PG) neuron terminals, increase the chemoafferent discharge. Additionally, vasoactive molecules produced within the CB may modulate hypoxic chemoreception by controlling blood flow and tissue PO2. Endothelin-1 (ET-1) increases basal CB chemosensory discharges in situ, probably due to its vasoconstrictor action. However, the actions of ET-1 on PG neurons or its expression in the PG are not known. Using immunohistochemistry, we found that endothelin-like peptides are expressed in the cat PG and CB under normoxic conditions. Exogenous applications of ET-1 increased the chemosensory activity in the vascularly perfused CB but were ineffective on either the CB or PG superfused preparations, both of which are devoid of vascular control. Thus, our data indicate that the excitatory effect of ET-1 in the carotid chemoreceptor system appears to be mainly due to a vasoconstrictor effect in the CB blood vessels. PMID- 16364258 TI - Functional compensation by other voltage-gated Ca2+ channels in mouse basal forebrain neurons with Ca(V)2.1 mutations. AB - Tottering (tg/tg) and leaner (tg(la)/tg(la)) mutant mice exhibit distinct mutations in the gene encoding the voltage-activated Ca(2+) channel alpha(1A) subunit (CACNA1A), the pore-forming subunit of the Ca(V)2.1 (P/Q type) Ca(2+) channels. These mice exhibit absence seizures and deficiencies in motor control and other functions. Previous work in cerebellar Purkinje neurons has shown that these mutations cause dramatic reductions in calcium channel function. Because Purkinje cell somata primarily express the Ca(V)2.1 channels, the general decrease in Ca(V)2.1 channel function is observed as a profound decrease in whole cell current. In contrast to Purkinje cells, basal forebrain (BF) neurons express all of the Ca(2+) channel alpha(1) subunits, with Ca(V)2.1 contributing approximately 30% to the whole-cell current in wild-type (+/+) mice. Here, we show that whole-cell Ba(2+) current densities in BF neurons are not reduced in the mutant genotypes despite a reduction in the Ca(V)2.1 contribution. By blocking the different Ca(2+) channel subtypes with specific pharmacological agents, we found a significant increase in the proportion of Ca(V)1 Ca(2+) current in mutant phenotypes. There was no change in tissue mRNA expression of calcium channel subtypes Ca(V)2.1, Ca(V)2.2, Ca(V)1.2, Ca(V)1.3, and Ca(V)2.3 in the tottering and leaner mutant mice. These results suggest that Ca(V)1 channels may functionally upregulate to compensate for reduced Ca(V)2.1 function in the mutants without an increase in Ca(v)1 message. Single-cell reverse transcription polymerase chain reaction (RT-PCR) experiments in a subset of sampled neurons revealed that approximately 90% of the cells could be considered cholinergic based on choline acetyltransferase (ChAT) mRNA expression. PMID- 16364259 TI - Brain-derived neurotrophic factor enhances conditioned taste aversion retention. AB - Brain-derived neurotrophic factor (BDNF) has recently emerged as one of the most potent molecular mediators of not only central synaptic plasticity, but also behavioral interactions between an organism and its environment. Our previous studies on the insular cortex (IC), a region of the temporal cortex implicated in the acquisition and storage of conditioned taste aversion (CTA), have demonstrated that induction of long-term potentiation (LTP) in the projection from the basolateral nucleus of the amygdala (Bla) to the IC, previous to CTA training, enhances the retention of this task. Recently, we found that intracortical microinfusion of BDNF induces a lasting potentiation of synaptic efficacy in the Bla-IC projection of adult rats in vivo. In this work, we present experimental data showing that intracortical microinfusion of BDNF previous to CTA training enhances the retention of this task. These findings support the concept that BDNF may contribute to memory-related functions performed by a neocortical area, playing a critical role in long-term synaptic plasticity. PMID- 16364260 TI - Purkinje cell activity during classical eyeblink conditioning in decerebrate guinea pigs. AB - Purkinje cells are the sole output from the cerebellar cortex and play a critical role during classical eyeblink conditioning. The present study revealed for the first time a learning-related change in individual Purkinje cell activity during successive eyeblink conditioning in decerebrate guinea pigs which permitted continuous single unit recording from the simplex lobe of the cerebellar cortex. The pair-conditioned group received paired presentation of the conditioned stimulus (CS) and unconditioned stimulus (US) until the frequency of the conditioned response (CR) exceeded 80%. The control group received a comparable number of the CS and US in a pseudorandom fashion. Responses of Purkinje cells to the CS were classified into four types: excitatory, inhibitory, a combination of the two, or no response. Approximately half of the recorded cells from both groups changed their response type at various conditioning stages. The firing frequency of a Purkinje cell to the CS showed a tendency to decrease in the pair conditioned group, while it had a tendency to increase in the pseudoconditioned group. This learning-related difference in change of response type was attributable to a difference in the change between the no response and the inhibitory response types. Correlation analysis of the temporal pattern between the neural activity and the CR revealed that most of the cells that developed an inhibitory response by paired conditioning acquired the CR-related temporal pattern. These results suggest that the learning-related Purkinje cells gain an inhibitory response with a temporal pattern correlated with the CR topography. PMID- 16364261 TI - Affective modulation of somatosensory-evoked potentials elicited by tactile stimulation. AB - The present investigation was aimed to evaluate the influence of emotional valence on brain correlates of non-painful somatosensory processing. For this purpose, we examined changes on the somatosensory-evoked potentials (SEP) elicited by frequent and deviant tactile stimuli (probability 14%) when subjects were viewing affective pictures. Twenty healthy volunteers aged between 19 and 47 years old participated in the study. The P50, N80, and P200 components of the SEP, as well as the P200 component of the visual-evoked potentials (VEP) elicited by the affective pictures were analyzed. Overall, a significant P50 amplitude reduction was observed when subjects were viewing unpleasant pictures, in comparison to pleasant pictures. Furthermore, larger SEP amplitudes were obtained in response to the deviant than to the frequent stimuli. In addition, unpleasant pictures elicited larger P200 amplitudes of the VEP than pleasant. Data suggest that affective stimuli may modulate the early processing of somatosensory information in the brain, probably reflecting the existence of an adaptive perceptual/attentional mechanism to motivationally relevant stimuli. PMID- 16364262 TI - Partial deletion of glial cell line-derived neurotrophic factor (GDNF) in mice: Effects on sucrose reward and striatal GDNF concentrations. AB - Glial cell line derived neurotrophic factor (GDNF) has been reported to alter the reward value of abused substances such as alcohol and cocaine as well as neural circuitry underlying reward. The role of GDNF in reward was further characterized in the present study using operant procedures to determine the value of a natural reward, sucrose, in GDNF heterozygous (GDNF+/-) mice versus wild-type (WT) mice. Female mice were tested for 2 h daily for 10 days in operant chambers with 2 levers. Responses on the correct lever allowed 5-s access to a dipper cup containing 15% sucrose. GDNF+/- and WT mice did not differ with acquisition or accuracy of responding. GDNF+/- mice emitted more responses than WT mice for sucrose, suggesting enhanced reward value of sucrose in these mice. In a separate experiment, concentrations of GDNF protein in striatal tissue were determined at 4, 8, and 12 months of age and found to be 38%-68% lower in GDNF+/- than WT mice at all three ages. Together, the results are consistent with an emerging literature indicating that reduced GDNF levels augment reward and increased GDNF levels attenuate reward, suggesting that GDNF plays an important role in neural systems mediating reward. PMID- 16364270 TI - Enzymatic preparation of maltohexaose, maltoheptaose, and maltooctaose by the preferential cyclomaltooligosaccharide (cyclodextrin) ring-opening reaction of Pyrococcus furiosus thermostable amylase. AB - Specific-length maltooligosaccharides, particularly maltohexaose, maltoheptaose, and maltooctaose, were prepared from cyclomaltooligosaccharides (cyclodextrins, CDs) by the preferential cyclodextrin ring-opening reaction of an amylolytic enzyme from Pyrococcus furiosus. The enzyme primarily produces maltohexaose, maltoheptaose, and maltooctaose by hydrolyzing alpha-, beta-, and gamma-CD, respectively. This study aims to develop a high-efficiency synthesis of specific maltooligosaccharides at high-purity. [formula: see text] PMID- 16364263 TI - Peptidic delta opioid receptor agonists produce antidepressant-like effects in the forced swim test and regulate BDNF mRNA expression in rats. AB - Systemically active, nonpeptidic delta opioid receptor agonists have been shown to produce antidepressant and anxiolytic effects in animal models in rodents. In addition, delta agonists have been shown to increase expression of brain-derived neurotrophic factor (BDNF) mRNA, an effect of some antidepressants, which may be important for the clinical efficacy of antidepressant drugs. The present study examined whether a variety of peptidic delta agonists, DPDPE, JOM-13, a systemically active derivative of DPDPE, deltorphin II, and H-Dmt-Tic-NH-CH2-Bid could produce convulsions and antidepressant-like effects in the forced swim test. In addition, some of these compounds were examined for their influence on BDNF mRNA expression. All four agonists dose-dependently decreased immobility in the forced swim test, indicating an antidepressant-like effect. Only JOM-13 produced convulsions at doses required for antidepressant-like effects. In addition, DPDPE increased BDNF mRNA expression, as measured by in situ hybridization, in the frontal cortex. The antidepressant-like effect of the agonists in the forced swim test and the increase in BDNF mRNA expression produced by DPDPE were blocked by the delta antagonist naltrindole. Therefore, activation of the delta receptor by centrally administered peptidic agonists and intravenously administered JOM-13 produces behavioral antidepressant-like effects without producing convulsions, and some peptidic agonists can increase BDNF mRNA expression, however, not as consistently as the systemically active nonpeptidic agonists. PMID- 16364271 TI - Acylation of carbohydrates over Al2O3: preparation of partially and fully acylated carbohydrate derivatives and acetylated glycosyl chlorides. AB - Selective and per-O-acylation of carbohydrate derivatives using acyl chlorides and Al2O3, a solid support reagent, is reported. This protocol does not require the addition of any base or activator. This methodology has been further extended to the selective acylation of carbohydrate diols and the one-pot preparation of acetylated glycosyl chlorides direct from free reducing sugars. The yields obtained in most of the cases are excellent. PMID- 16364272 TI - Syntheses of oligomannosides in solution and on a soluble polymer support: a comparison. AB - The alpha-(1-->6)-linked and the alpha-(1-->2)-linked linear mannotetraose glycosides and, respectively, and the branched mannopentaoside [R=CH2(CH2)2CH2Cl] were synthesised by conventional methods in solution, using trichloroacetimidate donors, and the products were obtained in 39%, 42% and 40% overall yield, respectively. For comparative purposes, the same two linear tetrasaccharides were prepared by use of MPEG as a soluble polymer support, the yields being 34% and 14%, respectively. An attempted MPEG-supported synthesis of the branched pentasaccharide was unsuccessful. The merits and shortcomings of oligosaccharide syntheses on MPEG are discussed. PMID- 16364273 TI - Rapid detection of common cytochrome P450 2D6 alleles in Caucasians. AB - CYP2D6 is a highly polymorphic enzyme that mediates the metabolism of around 20% of all currently prescribed drugs. Genetic variability within CYP2D6 results in poor (PM), intermediate (IM), extensive (EM) and ultra-rapid metabolisers (UM) of CYP2D6 substrates. Here we describe an assay which is able to detect the major PM (CYP2D6*3, *4, *5, *6), IM (CYP2D6*9, *10, *41) and UM (CYP2D6*nxn) alleles found in Caucasians. This assay is performed in two stages. The first stage is a multiplex long-range PCR which is used to simultaneously screen for whole gene deletions and duplications while isolating CYP2D6 from the CYP2D gene cluster to avoid pseudogene contamination. In the second stage, individuals with one or more copies of CYP2D6 are genotyped for PM and IM alleles using a two-tube multiplex Amplification Refractory Mutation System (ARMS). The specificity and reliability of the multiplex long-range PCR and subsequent ARMS were confirmed using a panel of positive controls that had been previously validated by PCR-RFLPs and DNA sequencing. This two-stage assay offers a robust and cheap alternative to many currently available CYP2D6 genotyping approaches. Our entire assay, once patient DNA has been extracted, can be run within 7 h using 10 microl PCRs. PMID- 16364274 TI - Nucleosome autoantibodies. AB - The nucleosome is a large protein-nucleic acid complex involved in DNA packing and in controlling genetic information. Under circumstances described below, this component, normally sequestered in the cell nucleus, is released into the extracellular milieu and then is easily accessible to cells of the immune system. For still not completely understood reasons, nucleosomes become immunogenic under particular conditions. Thus, anti-nucleosome autoantibodies (autoAb) have been described in connective tissue diseases and especially in systemic lupus erythematosus (SLE). This review describes the mechanisms leading to nucleosome production and anti-nucleosome autoimmunity, as well as the pathogenesis associated with nucleosomes. PMID- 16364275 TI - Relationship between plasma HDL subclasses distribution and lipoprotein lipase gene HindIII polymorphism in hyperlipidemia. AB - BACKGROUND: Different high-density lipoprotein (HDL) subclasses have distinct but interrelated metabolic functions. HDL directly influences the atherogenic process, and changes in HDL subclasses distribution may be related to the incidence and prevalence of atherosclerosis. Lipoprotein lipase (LPL) is an important enzyme for hydrolysis of triglyceride-rich lipoproteins, and its activity is positively correlated with the plasma HDL cholesterol level. LPL gene HindIII polymorphism has been found associated with variations in lipid levels, but the impact on HDL subclasses distribution is less clearly established. METHODS: The relative apolipoprotein (apo) A-I contents (% apoA-I) of plasma HDL subclasses were determined by two-dimensional gel electrophoresis coupled with immunodetection and LPL gene HindIII polymorphism was assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 173 hyperlipidemic and 155 normolipidemic subjects. RESULTS: The frequencies of 495TT genotype and allele T were the highest both in the hyperlipidemic and control groups. Compared with the control group, the frequency of 495TT genotype was higher, while the frequencies of 495TG and 495GG genotypes were significantly lower (P<0.05) in the hyperlipidemic group. Two-dimensional gel electrophoresis and immunodetection showed that HDL subclasses distribution was altered in hyperlipidemia, and had a general shift toward smaller size. Compared with the control group, the hyperlipidemic group had significantly higher relative apoA-I contents of prebeta1-HDL, prebeta2-HDL, HDL3b and HDL3a (P<0.05) and lower HDL2a and HDL2b levels (P<0.001). In the hyperlipidemic group, allele T carriers' frequency was higher than that in the control group (P<0.05), and the genotype of 495TT showed higher levels of plasma TG, apoB100, TG/HDL-C ratio, relative apoA-I contents of prebeta1-HDL, HDL3b and lower HDL2a, HDL2b compared with that of the 495GG genotype subgroup (P<0.05). In the control group, the genotype of 495TT had higher plasma TG, HDL3c and lower HDL2a compared with that of 495GG subgroup (P<0.05). CONCLUSIONS: The 495TT genotype of LPL gene HindIII polymorphism was associated with changes of HDL subclasses distribution in Chinese population with hyperlipidemia. The particle size of HDL shifted toward smaller, which, in turn, indicated that RCT might be weakened and HDL maturation might be abnormal in hyperlipidemic subjects with 495TT genotype. PMID- 16364276 TI - Postprandial lipemic response and lipoprotein composition in subjects with low or high cholesterol absorption efficiency. AB - BACKGROUND: The purpose of this study was to investigate the effect of differences in cholesterol absorption efficiency on the postprandial lipemia and lipoprotein composition. METHODS: Fifteen healthy subjects were divided into low and high cholesterol absorbers on the basis of serum cholestanol to cholesterol ratio. A high-performance liquid chromatographic method with evaporative light scattering detection was developed for quantitation of free and esterified cholesterol, triglycerides and major phospholipids from the same lipid extract in two runs utilizing the same internal standard. RESULTS: The free cholesterol to phosphatidylcholine ratio of chylomicrons was higher in the high cholesterol absorption group. The total increase of cholesterol in combined chylomicron and very low density lipoprotein (VLDL) fraction was also higher in this group. Chylomicron free cholesterol and cholesterol ester responses correlated with fasting low density lipoprotein (LDL) cholesterol. VLDL and VLDL1 triglyceride responses correlated inversely with fasting insulin and homeostasis model assessment of insulin resistance. CONCLUSIONS: High cholesterol absorption efficiency was seen in chylomicrons as higher cholesterol to phosphatidylcholine ratio during the postprandial peak. Chylomicron cholesterol response was linked to fasting LDL cholesterol and low VLDL triglyceride response to fasting insulin. PMID- 16364277 TI - Free fatty acid concentrations in gallbladder bile collected from Chilean patients with gallstones. AB - OBJECTIVES: To evaluate the association between gallstones and biliary free fatty acids (FFAs) as the first attempt to clarify whether biliary FFAs relate to developing gallbladder cancer (GBC) in Chile, which has the highest mortality rate in the world. DESIGN AND METHODS: Gallbladder bile from 21 male and 129 female patients with gallstones from Santiago was collected, and their FFAs were measured. The results were compared with those observed in a similar previous study performed in Niigata and Kochi Prefectures which showed the highest and lowest mortality rates, respectively, for GBC in Japan. RESULTS: Palmitoleic and linolenic acids compositions in Santiago patients were significantly lower than those in Niigata and Kochi patients, though these compositions in Niigata patients were significantly lower than those in Kochi patients. The levels of these FFAs compositions decreased in the order of Kochi, Niigata, and Santiago patients. CONCLUSIONS: Decreased biliary FFAs compositions may relate to developing gallstones and subsequently GBC. PMID- 16364278 TI - Information leakage from logically equivalent frames. AB - Framing effects are said to occur when equivalent frames lead to different choices. However, the equivalence in question has been incompletely conceptualized. In a new normative analysis of framing effects, we complete the conceptualization by introducing the notion of information equivalence. Information equivalence obtains when no choice-relevant inferences can be drawn from the speaker's choice of frame. We show that, to support the normative implications traditionally attributed to framing effects, frames must be equivalent in this sense. We also present new evidence for McKenzie and Nelson's (2003) reference point hypothesis, which posits a tendency to cast descriptions in terms of what has increased relative to the reference point. This leakage of information about relative state violates information equivalence, and gives rise to a normative account of the most robust finding in the attribute framing literature - the valence-consistency of preference shifts. We argue that, more generally, valenced descriptions leak information about perceived valence. Such "implicit recommendations" may generalize the reference point explanation of the valence-consistent shift. Normative and psychological implications of the information leakage framework are discussed. PMID- 16364279 TI - The acquisition of allophonic rules: statistical learning with linguistic constraints. AB - Phonological rules relate surface phonetic word forms to abstract underlying forms that are stored in the lexicon. Infants must thus acquire these rules in order to infer the abstract representation of words. We implement a statistical learning algorithm for the acquisition of one type of rule, namely allophony, which introduces context-sensitive phonetic variants of phonemes. This algorithm is based on the observation that different realizations of a single phoneme typically do not appear in the same contexts (ideally, they have complementary distributions). In particular, it measures the discrepancies in context probabilities for each pair of phonetic segments. In Experiment 1, we test the algorithm's performances on a pseudo-language and show that it is robust to statistical noise due to sampling and coding errors, and to non-systematic rule application. In Experiment 2, we show that a natural corpus of semiphonetically transcribed child-directed speech in French presents a very large number of near complementary distributions that do not correspond to existing allophonic rules. These spurious allophonic rules can be eliminated by a linguistically motivated filtering mechanism based on a phonetic representation of segments. We discuss the role of a priori linguistic knowledge in the statistical learning of phonology. PMID- 16364280 TI - What does syntax say about space? 2-year-olds use sentence structure to learn new prepositions. AB - Children as young as two use sentence structure to learn the meanings of verbs. We probed the generality of sensitivity to sentence structure by moving to a different semantic and syntactic domain, spatial prepositions. Twenty-six-month olds used sentence structure to determine whether a new word was an object category name (This is a corp!) or a spatial-relational term (This is acorp my box!). We argue that children rely on the intimate relationship between nouns in sentences and semantic arguments of predicate terms: Noting that a new word takes noun arguments identifies the new word as a predicate term, and directs the child's attention to relations among its arguments. PMID- 16364281 TI - Re-visiting the competence/performance debate in the acquisition of the counting principles. AB - Advocates of the "continuity hypothesis" have argued that innate non-verbal counting principles guide the acquisition of the verbal count list (Gelman & Galistel, 1978). Some studies have supported this hypothesis, but others have suggested that the counting principles must be constructed anew by each child. Defenders of the continuity hypothesis have argued that the studies that failed to support it obscured children's understanding of counting by making excessive demands on their fragile counting skills. We evaluated this claim by testing two , three-, and four-year-olds both on "easy" tasks that have supported continuity and "hard" tasks that have argued against it. A few noteworthy exceptions notwithstanding, children who failed to show that they understood counting on the hard tasks also failed on the easy tasks. Therefore, our results are consistent with a growing body of evidence that shows that the count list as a representation of the positive integers transcends pre-verbal representations of number. PMID- 16364282 TI - Design of a flexible platform for execution of medical decision support agents in the intensive care unit. AB - This paper addresses the design of a generic and scalable platform for the execution of medical decision support agents in the intensive care unit (ICU). As will be motivated, medical decision support agents can impose high computational load and in practical setups a large amount of such agents are typically running in parallel. Future ICU systems will rely on extensive medical decision support. However, in current systems only one workstation is typically dedicated for the execution of medical decision support agents. Therefore, we propose an architecture based on middleware technology to allow for easy distribution of the agents along multiple workstations. The architecture allows for easy integration with a general ICU data flow management architecture. PMID- 16364283 TI - Hex homeobox gene controls the transition of the endoderm to a pseudostratified, cell emergent epithelium for liver bud development. AB - Little is known about the mechanism by which embryonic liver, lung, and pancreas progenitor cells emerge from the endodermal epithelium to initiate organogenesis. Understanding this process and its genetic control provides insight into ontogeny, developmental abnormalities, and tissue regeneration. We find that shortly after hepatic endoderm cells are specified, they undergo a transition from a columnar, gut morphology to a pseudostratified morphology, with concomitant "interkinetic nuclear migration" (INM) during cell division. INM is a hallmark of pseudostratified epithelia and the process used by neural progenitors to emerge from the neural epithelium. We find that the transition of the hepatic endoderm, but not the neural epithelium, to a pseudostratified epithelium is dependent upon the cell-autonomous activity of the homeobox gene Hex. In the absence of Hex, hepatic endoderm cells survive but maintain a columnar, simple epithelial phenotype and ectopically express Shh and other genes characteristic of the midgut epithelium. Thus, Hex promotes endoderm organogenesis by promoting the transition to a pseudostratified epithelium, which in turn allows hepatoblasts to emerge into the stromal environment and continue differentiating. PMID- 16364284 TI - Zebrafish Foxd3 is required for development of a subset of neural crest derivatives. AB - foxd3 encodes a winged helix/forkhead class transcription factor expressed in the premigratory neural crest cells of many vertebrates. We have investigated the function of this gene in zebrafish neural crest by a loss of function approach using antisense morpholino oligonucleotides and immunostaining for Foxd3 protein. Knockdown of Foxd3 expression produces deficits in several differentiated neural crest derivatives, including jaw cartilage, peripheral neurons, and glia, and iridophore pigment cells. Other derivatives, such as melanophore and xanthophore pigment cells are not affected. Reduction in the expression of several lineage specific markers becomes evident soon after the onset of neural crest migration, suggesting that Foxd3 knockdown affects these lineages at early stages in their development. In contrast, analysis of the expression of early neural crest markers indicates little effect on neural crest induction or initial emigration. Finally, cell transplantation suggests that with respect to dorsal root ganglia neurons the Foxd3 requirement is cell autonomous, although Foxd3 itself is not detectable in differentiated DRG neurons. These results suggest that in zebrafish Foxd3 may not be required for induction of neural crest identity but is necessary for the differentiation of a subset of neural crest cell fates, perhaps in precursors of particular neural crest lineages. PMID- 16364285 TI - Mouse Rab23 regulates hedgehog signaling from smoothened to Gli proteins. AB - Sonic hedgehog (Shh) signaling is required for the growth and patterning of many tissues in vertebrate embryos, but important aspects of the Shh signal transduction pathway are poorly understood. For example, the vesicle transport protein Rab23 is a cell autonomous negative regulator of Shh signaling, but the process affected by Rab23 has not been defined. Here, we demonstrate that Rab23 acts upstream of Gli transcription factors in patterning neural cell types in the spinal cord. Double mutant analysis indicates that the primary target of Rab23 is the Gli2 activator and that Rab23 and Gli3 repressor have additive effects on patterning. Analysis of Gli3 protein suggests that Rab23 also has a role in promoting the production of Gli3 repressor. Although the membrane proteins Patched and Smoothened change subcellular localization in response to Shh, double mutant analysis demonstrates that Rab23 does not work through either Patched or Smoothened. Instead, Rab23 appears to regulate subcellular localization of essential components of the Hedgehog pathway that act downstream of Smoothened and upstream of Gli proteins. PMID- 16364286 TI - FoxA3 and goosecoid promote anterior neural fate through inhibition of Wnt8a activity before the onset of gastrulation. AB - Formation of the nervous system initially requires the acquisition of neural identity, which is achieved through the inhibition of epidermalizing factors. A regional patterning then takes place within the neural plate through the activity of caudalizing factors. These two processes are tightly regulated early in development by the dorsal organizer. Here, we show that, in zebrafish embryos, two transcription factors, FoxA3 and Goosecoid, coexpressed at the dorsal blastula margin, are required for the definition of anterior neural fate. Their inactivation results in deletions of anterior head structures associated with an increase of Wnt8 activity at the dorsal blastula margin. These phenotypes can be fully rescued by overexpression of Wnt inhibitors or by inactivation of wnt8a. Altogether, foxA3 and goosecoid cooperate to promote formation of anterior neural tissue by protecting, as early as blastula stage, presumptive anterior neural cells from an irreversible caudalization by the posteriorizing factor Wnt8a. PMID- 16364287 TI - Mobility assessment and validation of toxic elements in tunnel dust samples- subway and road using sequential chemical extraction and ICP-OES/GF AAS measurements. AB - The modified BCR three step sequential extraction procedure has been applied to homogenized urban dust samples collected in sedimentation chambers of two Prague tunnels (road, Letna, subway, and station Museum), and to SRM 1648 urban particulate matter (NIST) to validate the applicability of the procedure. Analyte concentrations in the fourth step were calculated from the total content of analytes determined and verified before, in case of the SRM 1648 certified values have been utilized. Concentrations of chosen elements As, Cd, Cr, Mn, Ni, Pb, Zn (Cu, Fe) in each fraction were measured by ICP-OES and GF AAS (arsenic determination) methods. The work has been focused on (i) levels investigation of toxic elements in dust samples and their distribution following a three-stage sequential extraction procedure, (ii) finding differences between two tunnel samples utilizing the estimation of mobility of trace elements, some macro component comparisons and main mineralogical phases determinations by XRD analysis, (iii) validation of the proposed procedure using SRM 1648 Urban Particulate Matter (NIST) and comparative data obtained by a capillary electrophoresis (CE) method. PMID- 16364288 TI - Glycyl-glutamine inhibits nicotine conditioned place preference and withdrawal. AB - Glycyl-glutamine (Gly-Gln) is an inhibitory dipeptide synthesized from beta endorphin(1-31). Previously, we showed that Gly-Gln inhibits morphine conditioned place preference, tolerance, dependence and withdrawal. In this study, we tested whether Gly-Gln's inhibitory activity extends to other rewarding drugs, specifically nicotine. Rats were conditioned with nicotine (0.6 mg/kg, s.c.) for four days and tested on day five. Glycyl-glutamine (100 nmol i.c.v.) inhibited acquisition and expression of a nicotine place preference significantly. Cyclo(Gly-Gln) (100 nmol i.c.v. or 25 mg/kg i.p.), a cyclic Gly-Gln derivative, blocked expression of nicotine place preference but Gly-d-Gln (100 nmol i.c.v.) was ineffective. To study nicotine withdrawal, rats were treated with nicotine (9 mg/kg/day) for seven days and conditioned place aversion was induced with mecamylamine (1 mg/kg, s.c.). Glycyl-glutamine blocked acquisition of place aversion to mecamylamine but not U50,488, a kappa opioid receptor agonist. Glycyl glutamine thus inhibits the rewarding effects of nicotine and attenuates withdrawal in nicotine dependent rats. PMID- 16364289 TI - Nitric oxide and its modulators in chronic constriction injury-induced neuropathic pain in rats. AB - This study was conducted to examine the role of nitric oxide (NO) in peripheral neuropathy induced by chronic constriction injury of sciatic nerve of rats by using NO precursor, NO donors and nitric oxide synthase (NOS) inhibitors. Chronic constriction injury of sciatic nerve of rats resulted in peripheral neuropathy as confirmed by nociceptive behavioural tests using mechanical, thermal and cold allodynia. NO precursor, L-arginine and NO donors sodium nitroprusside, S-nitroso N-acetylpenicillamine potentiated the hyperalgesia and allodynia significantly suggesting proalgesic effect in neuropathic rats. Intracerebroventricular (i.c.v.) administration of rats with NOS inhibitors such as L-N(G)-nitroarginine methyl ester, N-iminoethyl lysine and 7-nitroindazole did not show any effect but i.p. administration of NOS inhibitors aminoguanidine, L-N(G)-nitroarginine methyl ester and 7-nitroindazole caused alleviation of pain. The study confirms the involvement of endogenously synthesized and exogenously administered NO in chronic constriction injury-induced neuropathy in rats. Significant increase in the levels of nitrate and nitrite in ligated sciatic nerve suggest that local up regulation of NO in the production and maintenance of neuropathic pain. In conclusion, initial attempt to manipulate L-arginine: NO pathway is indicative of therapeutic potential of these interventions in the management of neuropathic pain. PMID- 16364290 TI - Mode of action of gingerols and shogaols on 5-HT3 receptors: binding studies, cation uptake by the receptor channel and contraction of isolated guinea-pig ileum. AB - Ginger (rhizomes of Zingiber officinale) has been shown to exert potent anti emetic properties, but its mode of action has not yet been elucidated. Among its active constituents, [6]-, [8]- and [10]-gingerol as well as [6]-shogaol were shown in different in vivo studies to be at least partly responsible for the drug's anti-emetic properties. In an attempt to gain more insight into the mode of action of these compounds, three different in vitro models were used to investigate their effects on 5-HT(3) receptors (serotonin receptor subtype) in more detail: [(14)C]guanidinium influx into N1E-115 cells which express 5-HT(3) receptors, isotonic contractions of the isolated guinea-pig ileum and equilibrium competition binding studies using a radioactively labeled 5-HT(3) receptor antagonist ([(3)H]GR65630) (3-(5-methyl-1H-imidazol-4-yl)-1-(1-methyl-1H-indol-3 yl)-1-propanone). All four compounds inhibited the [(14)C]guanidinium influx through 5-HT(3) receptor channels as well as contractions of the guinea-pig ileum induced by SR57227A ((4-amino)-(6-chloro-2-pyridyl)l-piperidine hydrochloride), a highly selective 5-HT(3) receptor agonist. Both effects were concentration dependent, with the following order of potency for both models: [6]-shogaol> or =[8]-gingerol>[10]-gingerol> or =[6]-gingerol. All compounds showed also weak anticholinergic and antineurokininergic activities in the guinea-pig ileum (acetylcholine and substance P are mediators of the 5-HT(3) receptor effect). The vanilloid receptor did not seem to be involved derived from experiments using capsazepine. None of the tested ginger substances, however, was able to displace [(3)H]GR65630 from its binding site (5-HT(3) receptor) neither on intact N1E-115 cells nor on the purified membranes of HEK-293 cells over-expressing the h5-HT(3) receptor. It may be concluded that [6]-, [8]-, [10]-gingerol and [6]-shogaol exert their anti-emetic effect at least partly by acting on the 5-HT(3) receptor ion-channel complex, probably by binding to a modulatory site distinct from the serotonin binding site. This may include indirect effects via receptors in the signal cascade behind the 5-HT(3) receptor channel complex such as substance P receptors and muscarinic receptors; this needs further investigation since ginger is effective against motion sickness which is cured by some vanilloids and by anticholinergics such as scopolamine. PMID- 16364291 TI - Gene structure, localization and role in oxidative stress of methionine sulfoxide reductase A (MSRA) in the monkey retina. AB - MSRA (EC 1.8.4.6) is a member of the methionine sulfoxide reductase family that can reduce methionine sulfoxide (MetO) in proteins. This repair function has been shown to protect cells against oxidative damage. In this study we have assembled the complete gene structure of msrA and identified the presence of two distinct putative promoters that generate three different transcripts. These transcripts were cloned by 5'RACE and code for three MSRA isoforms with different N-termini. The different forms of MSRA target to distinct intracellular regions. The main MSRA transcript (msrA1) had been previously shown to target the mitochondria. MsrA2 and 3 originate from a second promoter and target the cytosol and nuclei. In the monkey retina msrA message was detected mainly in the macular RPE-choroid region while its activity was measured mainly in the soluble fractions of fractionated neural retina and RPE-choroid. The MSRA protein is found throughout the retina but is especially abundant at the photoreceptor synapses, ganglion and Muller cells. Interestingly, MSRA was not detected in the mitochondria of the photoreceptor inner segments. The RPE in the peripheral retina shows very low levels of expression but the RPE in the macular region is strongly labeled. Targeted silencing of msrA message rendered cultured RPE cells more sensitive to oxidative damage suggesting a role for MSRA in RPE protection against oxidative stress. Collectively these data suggest MSRA may play an important role in protecting macular RPE from oxidative damage. PMID- 16364292 TI - Hypoxia preconditioning protects corneal stromal cells against induced apoptosis. AB - The purpose of this study, was to determine whether hypoxia preconditioning can protect corneal stromal cells from UV stress and cytokine mediated apoptosis. Two models were implemented. First, primary cultured bovine corneal fibroblasts were preconditioned with 0.5-1.5% O2 for 4 hr and stressed with UV-irradiation or stimulation of Fas receptor. Second, bovine eyes were preconditioned with 0.5% O2 for 4 hr and stressed by epithelial scraping to induce anterior keratocyte apoptosis. Cell fate was analyzed at 4 hr after stress using quantitative TUNEL or condensed nuclei assays. Cell apoptotic rates in hypoxia preconditioned groups were significantly lower (50-80%) than that of normoxia control groups. Hypoxia prevented the degradation of the transcription factor HIF-1alpha. CoCl2 (100-200 microM), a chemical inducer of HIF-1alpha, also produced strong protection against UV and Fas induced apoptosis. Moreover, hypoxia preconditioned media protected cells against UV-induced apoptosis. These findings demonstrate that hypoxia preconditioning has a generalized protective effect against stromal fibroblast and keratocyte apoptosis and suggest that HIF-1alpha mediated expression and secretion of protective factors is involved. PMID- 16364293 TI - Photooxidation of A2-PE, a photoreceptor outer segment fluorophore, and protection by lutein and zeaxanthin. AB - A2-PE is a pigment that forms as a byproduct of the visual cycle, its synthesis from all-trans-retinal and phosphatidylethanolamine occurring in photoreceptor outer segments. A2-PE is deposited in retinal pigment epithelial (RPE) cells secondary to phagocytosis of shed outer segment membrane and it undergoes hydrolysis to generate the RPE lipofuscin fluorophores, A2E, iso-A2E and other minor cis-isomers of A2E. We have demonstrated that A2-PE can initiate photochemical processes that involve the oxidation of A2-PE and that, by analogy with A2E are likely to include the formation of reactive moieties. We also show that potential sources of protection against the photooxidation of A2-PE are the lipid-soluble carotenoids zeaxanthin and lutein (xanthophylls), that constitute the yellow pigment of the macula. Irradiation of A2-PE in the presence of lutein or zeaxanthin suppressed A2-PE photooxidation and in experiments in which we compared the antioxidant capability of zeaxanthin and lutein to alpha-tocopherol, the carotenoids were more potent. Additionally, the effect with zeaxanthin was consistently more robust than with lutein and when alpha-tocopherol was combined with either carotenoid, the outcome was additive. Lutein, zeaxanthin and alpha tocopherol were all efficient quenchers of singlet oxygen. We have also shown that lutein and zeaxanthin can protect against A2-PE/A2E photooxidation without appreciable consumption of the carotenoid by chemical reaction. This observation contrasts with the pronounced susceptibility of A2E and A2-PE to photooxidation and is of interest since lutein, zeaxanthin, A2E and A2-PE all have conjugated systems of carbon-carbon double bonds terminating in cyclohexenyl end-groups. The structural features responsible for the differences in quenching mechanisms are discussed. It has long been suspected that macular pigment protects the retina both by filtering high-energy blue light and by serving an antioxidant function. Evidence presented here suggests that the photochemical reactions against which lutein and zeaxanthin protect, may include those initiated by the A2-PE. Quantitative HPLC analysis revealed that in eyecups of C57BL/6J and BALB/cByJ mice, levels of A2-PE were several fold greater than the cleavage product, A2E. Taken together, these results may have implications with respect to the involvement of A2-PE formation in mechanisms underlying blue light-induced photoreceptor cell damage and may be significant to retinal degenerative disorders, such as those associated with ABCA4 mutations, wherein there is a propensity for increased A2-PE synthesis. PMID- 16364294 TI - Focus on Molecules: Pigment epithelium-derived factor (PEDF). PMID- 16364295 TI - Retinal vessel dilation following repletion of vitamin A deficiency. AB - We hypothesized that because depletion of vitamin A blocks the initiation of phototransduction, such inhibition of functional activation should lead to decrease retinal metabolism and perfusion. In a case study of a vitamin A depleted patient, we found that retinal vessel diameters, a surrogate measure of retinal perfusion, increased in concert with the restitution of electroretinographic function following vitamin A supplementation. When normalized to conditions after treatment, the relative magnitude of study parameters at presentation were: scotopic electroretinography B-wave amplitude 1.2%, photopic electroretrinography B-wave amplitude 23%, retinal vein diameter 88%, retinal artery diameter 94%. These observations support that activation of the visual process results in increased retinal metabolism and perfusion. PMID- 16364296 TI - The expression of advanced glycation endproduct receptors in rpe cells associated with basal deposits in human maculas. AB - Basal deposits within Bruch's membrane are associated with aging and age-related macular degeneration (AMD) although the factors causing their formation are incompletely understood. Advanced glycation endproducts (AGEs) accumulate in Bruch's membrane including basal deposits and drusen with aging. One mechanism by which AGEs alter a cell's phenotype is via AGE receptors. The purpose of this study was to immunolocalize and quantify the expression of AGE receptors by RPE cells associated with basal deposits or normal Bruch's membrane that were microdissected from human maculas. Postmortem eyes from 14 aged control donors and five donors with non-neovascular AMD were cryopreserved. RPE cells associated with normal Bruch's membrane or basal deposits were laser capture microdissected. The RNA was extracted and used for RT-qPCR to quantify the expression of RAGE, AGE R1, AGE R2, and AGE R3. Streptavidin alkaline phosphatase immunohistochemistry for these receptors was also performed and sections were bleached from 14 normal and nine AMD donors. RT-qPCR showed significant upregulation of RAGE, AGE R1, and AGE R3 in RPE cells overlying basal deposits compared to cells attached to morphologically normal Bruch's membrane. Immunohistochemical analysis for RAGE, AGER1, R2, and R3 showed diffuse, light staining of RPE cells and strong choriocapillaris staining in areas of normal Bruch's membrane. In areas of basal deposits, the RPE had more intense staining for RAGE and AGER1 compared to regions of normal Bruch's membrane. These results suggest that AGE receptors could influence the formation of basal deposits during aging and AMD. PMID- 16364297 TI - Upregulation of RAGE and its ligands in proliferative retinal disease. AB - We sought to study the presence of the receptor for advanced glycation endproducts (RAGE) and its ligands, advanced glycation endproducts (AGEs), S100/calgranulins and amphoterin (high mobility group box 1 protein; HMGB1), in the vitreous cavity and epiretinal membranes (ERMs) of eyes of patients with proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR). Undiluted vitreous specimens were collected from 30 eyes of 30 patients undergoing pars plana vitrectomy for repair of retinal detachment (RD) secondary to PDR (n = 15) or PVR (n = 15). The vitreous samples obtained from 10 eyes undergoing macular hole repair were used as controls. Epiretinal membranes were obtained from eight eyes with PDR and from 10 eyes with PVR. The levels of AGEs in the vitreous were measured using ELISA. The vitreous levels of soluble RAGE (sRAGE), S100/calgranulins and amphoterin were measured using Western blot analyses. The localization of RAGE and its ligands in ERMs was determined with immunohistochemistry. The vitreous levels of sRAGE were significantly increased in both PDR and PVR (p < or = 0.05) compared to control vitreous. In both PDR and PVR, the vitreous levels of AGEs (p < or = 0.01), S100/calgranulins (p < or = 0.05), and amphoterin (p < or = 0.01) were also elevated compared to control eyes. Expression of RAGE was detected in six of eight ERMs from eyes with PDR and eight of 10 ERMs from eyes with PVR. Many cells expressing RAGE also expressed vimentin, suggesting a glial cell origin. Ligands for RAGE were also detected in ERMs, with AGEs detected in five eyes with PDR and eight eyes with PVR. Similarly, S100 and amphoterin ERM expression was observed in six eyes with PDR; these ligands were also expressed in ERMs from eyes with PVR (8 and 7 cases, respectively). We conclude that RAGE and its ligands are increased in the vitreous cavity of eyes with PDR and PVR and are present in ERMs of eyes with these proliferative retinal disorders. These findings suggest a role for the proinflammatory RAGE axis in the pathogenesis of proliferative retinal diseases. PMID- 16364298 TI - Cerebral neurons of transgenic ALS mice are vulnerable to glutamate release stimulation but not to increased extracellular glutamate due to transport blockade. AB - Mechanisms of motor neuron loss in amyotrophic lateral sclerosis (ALS) are unknown, but it has been postulated that excitotoxicity due to excessive glutamatergic neurotransmission by decreased efficiency of glutamate transport may be involved in both familial (FALS) and sporadic ALS. Using microdialysis in vivo, we tested the effects of the glutamate transport inhibitor L-trans pyrrolidine-2,4-dicarboxylate (PDC) and of 4-aminopyridine (4-AP), which stimulates glutamate release from nerve endings, in the hippocampus and motor cortex of wild type (WT) and transgenic SOD1/G93A mice, an established model of FALS. Perfusion of 4-AP induced convulsions, expression of the inducible stress marker heat-shock protein 70 (HSP70) and hippocampal neuronal loss. These effects were similar in both WT and G93A mice, and, in both groups, they were prevented by the previous systemic administration of the NMDA receptor antagonist MK-801. In contrast, perfusion of PDC resulted in a large and long-lasting (2 h) increase of extracellular glutamate, but no convulsions, neuronal damage or HSP70 expression were observed in either the WT or the G93A mice. Our results demonstrate that SOD1 G93A mutation does not enhance the vulnerability to endogenous glutamate-mediated excitotoxicity in brain, neither by blocking glutamate transport nor by stimulating its release. Therefore, these data do not support the possibility that glutamate transport deficiency may be an important factor of brain neuronal degeneration in familial ALS. PMID- 16364299 TI - Dietary curcumin counteracts the outcome of traumatic brain injury on oxidative stress, synaptic plasticity, and cognition. AB - The pervasive action of oxidative stress on neuronal function and plasticity after traumatic brain injury (TBI) is becoming increasingly recognized. Here, we evaluated the capacity of the powerful antioxidant curry spice curcumin ingested in the diet to counteract the oxidative damage encountered in the injured brain. In addition, we have examined the possibility that dietary curcumin may favor the injured brain by interacting with molecular mechanisms that maintain synaptic plasticity and cognition. The analysis was focused on the BDNF system based on its action on synaptic plasticity and cognition by modulating synapsin I and CREB. Rats were exposed to a regular diet or a diet high in saturated fat, with or without 500 ppm curcumin for 4 weeks (n = 8/group), before a mild fluid percussion injury (FPI) was performed. The high-fat diet has been shown to exacerbate the effects of TBI on synaptic plasticity and cognitive function. Supplementation of curcumin in the diet dramatically reduced oxidative damage and normalized levels of BDNF, synapsin I, and CREB that had been altered after TBI. Furthermore, curcumin supplementation counteracted the cognitive impairment caused by TBI. These results are in agreement with previous evidence, showing that oxidative stress can affect the injured brain by acting through the BDNF system to affect synaptic plasticity and cognition. The fact that oxidative stress is an intrinsic component of the neurological sequel of TBI and other insults indicates that dietary antioxidant therapy is a realistic approach to promote protective mechanisms in the injured brain. PMID- 16364301 TI - Effects of post-ingestion and physical conditions on PCR amplification of host blood meal DNA in mosquitoes. AB - The effects of post-ingestion and physical conditions under which killed mosquitoes are stored on the success of detecting blood meal DNA of Anopheles stephensi and Culex quinquefasiatus were investigated by polymerase chain reaction (PCR) amplification at the human mitochondrial DNA cytochrome b (cytB) gene. Host DNA extracted from the blood meal up to 33 h post-ingestion in both species acts as an efficient template for PCR amplification. However, more DNA concentration is needed for meals digested for a longer time, and successful PCR amplification from meals digested for 36 h,dropped to a faint band. There were no differences between PCR success rate for samples stored at +4 or -20 degrees C, but less successful products were observed in samples kept at 4 degrees C for the periods longer than 30 h digestion. The results of this study are important for conducting malaria epidemiological studies that provide information about the degree of contact between human hosts and mosquito vectors, impact of vector controls such as bed nets and repellents, and the transmission dynamics of human malaria and other vector-borne diseases. PMID- 16364302 TI - P70 S6 kinase mediates tau phosphorylation and synthesis. AB - Currently, we found that the 70-kDa p70 S6 kinase (p70S6K) directly phosphorylates tau at S262, S214, and T212 sites in vitro. By immunoprecipitation, p-p70S6K (T421/S424) showed a close association with p-tau (S262 and S396/404). Zinc-induced p70S6K activation could only upregulate translation of total S6 and tau but not global proteins in SH-SY5Y cells. The requirement of p70S6K activation was confirmed in the SH-SY5Y cells that overexpress wild-type htau40. Level of p-p70S6K (T421/S424) was only significantly correlated with p-tau at S262, S214, and T212, but not T212/S214, in Alzheimer's disease (AD) brains. These suggested that p70S6K might contribute to tau related pathologies in AD brains. PMID- 16364300 TI - Plasmodium falciparum: Rosettes do not protect merozoites from invasion inhibitory antibodies. AB - Rosetting is a parasite adhesion phenotype associated with severe malaria in African children. Why parasites form rosettes is unknown, although enhanced invasion or immune evasion have been suggested as possible functions. Previous work showed that rosetting does not enhance parasite invasion under standard in vitro conditions. We hypothesised that rosetting might promote invasion in the presence of host invasion-inhibitory antibodies, by allowing merozoites direct entry into the erythrocytes in the rosette and so minimising exposure to plasma antibodies. We therefore investigated whether rosetting influences invasion in the presence of invasion-inhibitory antibodies to MSP-1. We found no difference in invasion rates between isogenic rosetting and non-rosetting lines from two parasite strains, R29 and TM284, in the presence of MSP-1 antibodies (P = 0.62 and P = 0.63, Student's t test, TM284 and R29, respectively). These results do not support the hypothesis that rosettes protect merozoites from inhibitory antibodies during invasion. The biological function of rosetting remains unknown. PMID- 16364303 TI - C-terminal truncation modulates both nucleation and extension phases of tau fibrillization. AB - Proteolytic post-translational modification has been proposed as an early stage event in the aggregation of tau protein and formation of neurofibrillary lesions in Alzheimer's disease. Caspases and other proteases cleave tau in vivo at discrete locations including Asp421 and Glu391. Both cleavage products are prone to aggregation relative to wild-type, full-length tau protein. To determine the mechanism underlying this effect, the fibrillization of tau truncated after Asp421 and Glu391 residues was characterized in a full-length four-repeat tau background using quantitative electron microscopy methods under homogeneous nucleation conditions. Both C-terminal truncations decreased critical concentration relative to full-length tau, resulting in more filament mass at reaction plateau. Moreover, truncation directly augmented the efficiency of the nucleation reaction. The results suggest the mechanism through which C-terminal proteolysis can modulate tau filament accumulation depending on whether it precedes or follows nucleation. PMID- 16364304 TI - Molecular determinants for interaction of SHEP1 with Cas localize to a highly solvent-protected region in the complex. AB - Protein-protein interactions between SHEP and Cas proteins influence cellular signaling through tyrosine kinases, as well as integrin-mediated signaling, and may be linked to antiestrogen resistance. Data from past studies suggests that association between SHEP and Cas proteins is critical for these cellular effects. In this study, the interacting domains of each protein were co-expressed in bacteria and a soluble stable complex was purified. Deuterium exchange mass spectrometry was used to define regions that are buried when SHEP1 is in complex with Cas. The results reveal four segments in SHEP1 that are highly protected, including a region (residues 619-640) that contains a key residue, tyrosine 635, required for association with Cas. This region is predominately hydrophilic, yet remains protected from solvent in the complex. PMID- 16364306 TI - RNA editing sites in plant mitochondria can share cis-elements. AB - RNA editing in flowering plant mitochondria alters numerous C nucleotides in a given mRNA molecule to U residues. To investigate whether neighbouring editing sites can influence each other we analyzed in vitro RNA editing of two sites spaced 30 nt apart. Deletion and competition experiments show that these two sites carry independent essential specificity determinants in the respective upstream 20-30 nucleotides. However, deletion of a an upstream sequence region promoting editing of the upstream site concomitantly decreases RNA editing of the second site 50-70 nucleotides downstream. This result suggests that supporting cis-/trans-interactions can be effective over larger distances and can affect more than one editing event. PMID- 16364305 TI - The membrane anchors of the heme chaperone CcmE and the periplasmic thioredoxin CcmG are functionally important. AB - The cytochrome c maturation system of Escherichia coli contains two monotopic membrane proteins with periplasmic, functional domains, the heme chaperone CcmE and the thioredoxin CcmG. We show in a domain swap experiment that the membrane anchors of these proteins can be exchanged without drastic loss of function in cytochrome c maturation. By contrast, the soluble periplasmic forms produced with a cleavable OmpA signal sequence have low biological activity. Both the chimerical CcmE (CcmG'-'E) and the soluble periplasmic CcmE produce low levels of holo-CcmE and thus are impaired in their heme receiving capacity. Also, both forms of CcmE can be co-precipitated with CcmC, thus restricting the site of interaction of CcmE with CcmC to the C-terminal periplasmic domain. However, the low level of holo-CcmE formed in the chimera is transferred efficiently to cytochrome c, indicating that heme delivery from CcmE does not involve the membrane anchor. PMID- 16364308 TI - Mitochondrial targeting of growth suppressor protein DLC2 through the START domain. AB - Deleted in liver cancer 2 (DLC2) is a candidate tumor suppressor frequently found to be deleted in hepatocellular carcinoma. In this study, we determined the subcellular localization of DLC2. Co-localization and biochemical fractionation studies revealed that DLC2 localized to mitochondria. In addition, the DLC2 containing cytoplasmic speckles were in proximity to lipid droplets. A DLC2 mutant containing the steroidogenic acute regulatory protein-related lipid transfer (START) domain only showed a localization pattern identical to that of DLC2. Taken together, we have provided the first evidence for mitochondrial localization of DLC2 through the START domain. These findings might have implications in liver physiology and carcinogenesis. PMID- 16364307 TI - The identification of a functional interaction between PKC and topoisomerase II. AB - Topoisomerase II plays an essential role in the segregation of chromosomes during cell division. It is also a major component of the nuclear matrix. Proteins that interact with and regulate this essential enzyme are of great interest. To investigate the role of proteins interacting with the N-terminal domain of the Saccharomyces cerevisiae topoisomerase II, we used a yeast two-hybrid protein interaction screen. We identified an interaction between the catalytic domain of the yeast protein kinase 1 enzyme (Pkc1) and the N-terminal domain of the S. cerevisiae topoisomerase II. The S. cerevisiae Pkc1 is the homologue of the mammalian calcium dependent PKC. PMID- 16364309 TI - ABC transporters involved in the transport of plant secondary metabolites. AB - Plants produce a large number of secondary metabolites, such as alkaloids, terpenoids, polyphenols, quinones and many further compounds having combined structures of those groups. Physiological roles of those metabolites for plants are still under investigation, but they play, at least in part, important functions as protectants for plant bodies against herbivores and pathogens, as well as from physical stresses like ultraviolet light and heat. In order to accomplish these functions, biosyntheses and accumulation of secondary metabolites are highly regulated in a temporal and spatial manner in plant organs, where they can appropriately accumulate. In this mini-review, I introduce the mechanism of accumulation and membrane transport of these metabolites, in particular, focusing on ATP-binding cassette transporters involved. PMID- 16364310 TI - Crystal structure of a plant immunophilin domain involved in regulation of MDR type ABC transporters. AB - We present the three-dimensional structure of the N-terminal FK506-binding protein (FKBP)-like domain of the immunophilin FKBP42 from Arabidopsis thaliana. The data provide the structural background for the explanation of key functional properties reported previously. PMID- 16364311 TI - Structure of the PRYSPRY-domain: implications for autoinflammatory diseases. AB - We determined the first structure of PRYSPRY, a domain found in over 500 different proteins, involved in innate immune signaling, cytokine signaling suppression, development, cell growth and retroviral restriction. The fold encompasses a 7-stranded and a 6-stranded antiparallel beta-sheet, arranged in a beta-sandwich. In the crystal, PRYSPRY forms a dimer where the C-terminus of an acceptor molecule binds to the concave surface of a donor molecule, which represents a putative interaction site. Mutations in the PRYSPRY domains of Pyrin, which are responsible for familial Mediterranean fever, map on the putative PRYSPRY interaction site. PMID- 16364312 TI - Identification of otubain 1 as a novel substrate for the Yersinia protein kinase using chemical genetics and mass spectrometry. AB - Yersinia encodes a protein kinase, YpkA, which disrupts the actin cytoskeleton. Using an approach termed chemical genetics, we identified a 36-kDa substrate for YpkA in both J774 lysates and bovine brain cytosol. Mass spectrometry analysis identified this substrate as FLJ20113, an open reading frame that corresponds to otubain 1, a deubiquitinating enzyme implicated in immune cell clonal anergy. We demonstrate that otubain 1 is phosphorylated by YpkA in vitro and interacts with YpkA and actin in vivo. Identification of otubain 1 as a YpkA substrate suggests that regulation of immune cell anergy may be a survival mechanism for Yersinia. PMID- 16364313 TI - Activation of pannexin 1 channels by ATP through P2Y receptors and by cytoplasmic calcium. AB - The ability for long-range communication through intercellular calcium waves is inherent to cells of many tissues. A dual propagation mode for these waves includes passage of IP3 through gap junctions as well as an extracellular pathway involving ATP. The wave can be regenerative and include ATP-induced ATP release via an unknown mechanism. Here, we show that pannexin 1 channels can be activated by extracellular ATP acting through purinergic receptors of the P2Y group as well as by cytoplasmic calcium. Based on its properties, including ATP permeability, pannexin 1 may be involved in both initiation and propagation of calcium waves. PMID- 16364314 TI - Modulation of MPP+ uptake by procyanidins in Caco-2 cells: involvement of oxidation/reduction reactions. AB - It is becoming increasingly evident that the absorption of certain nutrients and drugs and their effects are largely influenced by the concomitant ingestion of other substances. As various xeno- and endobiotics belong to the class of organic cations, the aim of this work was to study the modulation of the intestinal apical uptake of organic cations by diet procyanidins. Five procyanidin fractions with different structural complexity were obtained after fractionation of a grape seed extract. The effect of these compounds on 1-methyl-4-phenylpyridinium (MPP+) uptake was evaluated in Caco-2 cells. Apical uptake of 3H-MPP+ by Caco-2 cells was increased by a 60 min exposure to 600 microg ml(-1) of procyanidin fractions, that increase being positively related with procyanidins structural complexity. It was verified that 3H-MPP+ uptake increased with preincubation time. It was speculated that procyanidins were oxidized during preincubation, this change could interfered with transport activity. Tested oxidizing agents showed that the redox state of the transporter could affect its activity. Additionally, trans stimulation experiments showed that catechin and fraction I (the simpler fraction) can use the same transporter as MPP+. The results are compatible with the hypothesis of these compounds being competitive inhibitors of MPP+ transport. In conclusion, procyanidins are capable to modulate MPP+ apical uptake in Caco-2 cells, this transport being most probably modulated through oxidation-reduction phenomena. Interactions between these compounds and drugs present in the diet may affect their absorption and bioavailability. Both the concentration and complexity of the procyanidin compounds should be taken into account in medical practice. PMID- 16364315 TI - Human and yeast Hsp110 chaperones exhibit functional differences. AB - Hsp110 proteins constitute a heterogeneous family of abundant molecular chaperones, related to the Hsp70 proteins and exclusively found in the cytosol of eukaryotic organisms. Hsp110 family members are described as efficient holdases, preventing the aggregation and assisting the refolding of heat-denatured model substrates in the presence of Hsp70 chaperones and their co-chaperones. To gain more insights into the mode of action of this protein family we compared two homologues representing two subtypes of Hsp110 proteins, S. cerevisiae Sse1 and H. sapiens Apg-2, in their structural and functional properties in vitro. In contrast to previous publications both proteins exhibited intrinsic ATPase activities, which only in the case of Sse1 could be stimulated by the Hsp40 co chaperone Sis1. Similar to Hsp70 proteins ATP binding and hydrolysis induced conformational rearrangements in both Hsp110 proteins as detected by tryptophane fluorescence. However, nucleotide induced changes in the proteolytic digestion pattern were detected only for Sse1. Sse1 and Apg-2 thus show significant differences in their biochemical properties, which may relate to differences in their functional roles in vivo. PMID- 16364316 TI - Improved gating of a chimeric alpha7-5HT3A receptor upon mutations at the M2-M3 extracellular loop. AB - Acetylcholine-evoked currents of the receptor chimera alpha7-5HT3A V201 expressed in Xenopus oocytes are strikingly small when compared to the amount of alpha bungarotoxin binding sites detected at the oocyte membrane. Since the chimeric receptor is made of the extracellular N-terminal region of the rat alpha7 nicotinic acetylcholine receptor and the C-terminal region of the mouse 5-HT3A receptor, which includes the ion channel, we hypothesized that communication between these two regions was not optimal. Here, we show that mutating to aspartate several adjacent positions in the M2-M3 extracellular linker increases current amplitudes to different extents, thus confirming the important role of this region on receptor gating. PMID- 16364317 TI - The structure of the third intracellular loop of the muscarinic acetylcholine receptor M2 subtype. AB - We have examined whether the long third intracellular loop (i3) of the muscarinic acetylcholine receptor M2 subtype has a rigid structure. Circular dichroism (CD) and nuclear magnetic resonance spectra of M2i3 expressed in and purified from Escherichia coli indicated that M2i3 consists mostly of random coil. In addition, the differential CD spectrum between the M2 and M2deltai3 receptors, the latter of which lacks most of i3 except N- and C-terminal ends, gave no indication of secondary structure. These results suggest that the central part of i3 of the M2 receptor has a flexible structure. PMID- 16364318 TI - Cathepsin B and its interacting proteins, bikunin and TSRC1, correlate with TNF induced apoptosis of ovarian cancer cells OV-90. AB - Increasing evidence suggests that lysosomal cysteine proteases cathepsins contribute to the progression of cell apoptosis. Here we found that apoptosis of ovarian cancer cells OV-90 triggered by TNF was cathepsin B-depended. Two cathepsin B binding proteins, bikunin and TSRC1, were identified by yeast two hybrid method and the interactions were confirmed in vitro and in vivo. Overexpression of bikunin could suppress TNF-induced apoptosis of OV-90 cells, and TSRC1 overexpression had an opposite effect on apoptosis. The presented results suggest that cathepsin B and its interacting proteins, bikunin and TSRC1, are involved in the apoptotic pathway of ovarian cancer cells. PMID- 16364319 TI - Transformation-associated gene regulation by ATF6alpha during hepatocarcinogenesis. AB - We have previously reported that the endoplasmic reticulum (ER) stress-regulated transmembrane transcription factor 6 alpha (ATF6alpha) is implicated in the pathogenesis of hepatocellular carcinomas (HCCs). In order to further identify genes that are regulated by ATF6alpha, the global gene expression profiles of the ATF6alpha-transfected and untransfected HCC cell line, HLF, were analyzed. These results were then compared with the differential gene expression patterns of poorly differentiated HCC and control non-tumorous liver tissue. Our findings demonstrate that at least 18 genes are specifically upregulated by ATF6alpha, while another UPR mediator, XBP1 or ER-stress inducer, thapsigargin could partially stimulate or even repress some of them in HCC cells. Moreover, six of these identified genes contain potential ER stress-responsive elements and/or unfolded protein response elements in their 5' regulatory regions. PMID- 16364320 TI - Crystal structure of Escherichia coli SufC, an ABC-type ATPase component of the SUF iron-sulfur cluster assembly machinery. AB - SufC is an ATPase component of the SUF machinery, which is involved in the biosynthesis of Fe-S clusters. To gain insight into the function of this protein, we have determined the crystal structure of Escherichia coli SufC at 2.5A resolution. Despite the similarity of the overall structure with ABC-ATPases (nucleotide-binding domains of ABC transporters), some key differences were observed. Glu171, an invariant residue involved in ATP hydrolysis, is rotated away from the nucleotide-binding pocket to form a SufC-specific salt bridge with Lys152. Due to this salt bridge, D-loop that follows Glu171 is flipped out to the molecular surface, which may sterically inhibit the formation of an active dimer. Thus, the salt bridge may play a critical role in regulating ATPase activity and preventing wasteful ATP hydrolysis. Furthermore, SufC has a unique Q-loop structure on its surface, which may form a binding site for its partner proteins, SufB and/or SufD. PMID- 16364321 TI - Physical and functional interactions between STAT3 and Kaposi's sarcoma associated herpesvirus-encoded LANA. AB - The Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded latency-associated nuclear antigen (LANA) is known to modulate viral and cellular gene expression. We show that LANA directly associates with an interleukin-6 signal transducer, signal transducer and activator of transcription 3 (STAT3) and that LANA enhances the transcriptional activity of STAT3. Coimmunoprecipitation studies documented a physical interaction between LANA and STAT3 in transiently transfected 293T cells as well as the KSHV-infected primary effusion lymphoma (PEL) cell line. Furthermore, small-interfering RNA-mediated reduction of LANA expression decreased the STAT3-dependent transcription in KSHV-positive PEL cells, whereas overexpression of LANA enhanced STAT3 activity in KSHV-negative B lymphoma cells. These data demonstrate that LANA is a transcriptional co-activator of STAT3, and may have implications for the pathogenesis of KSHV-associated diseases. PMID- 16364322 TI - Expression of yeast transcriptional activator MSN1 promotes accumulation of chromium and sulfur by enhancing sulfate transporter level in plants. AB - MSN1 is a putative yeast transcriptional activator involved in chromium (Cr) accumulation. Here we show that overexpression of MSN1 enhances Cr and sulfur accumulation and Cr tolerance in transgenic tobacco. In addition, we found that expression of NtST1 (Nicotiana tabacum sulfate transporter 1) was elevated in MSN1- expressing transgenic tobacco, suggesting that chromate and sulfate are taken up via the sulfate transporter in plants. Supporting this, expression of NtST1 increased levels of Cr and S in Saccharomyces cerevisiae. Our findings suggest that yeast transcriptional activators can be used for developing effective metal remediators, and for improving the nutritional status of plants. PMID- 16364323 TI - Patterns of proopiomelanotropin and proopiocortin gene expression and of immunohistochemistry for gonadotropin-releasing hormones (lGnRH-I and III) during the life cycle of a nonparasitic lamprey: relationship to this adult life history type. AB - There are two adult life history types among lamprey species, nonparasitic and parasitic, with the former commencing the final interval of sexual maturation immediately after metamorphosis. There are no extensive studies that directly compare hormone profiles during the life cycles of nonparasitic and parasitic lamprey species, yet such data may explain differences in development, reproductive maturation, and feeding status. The present study uses immunohistochemistry to show the life cycle profiles for gonadotropin-releasing hormones (GnRH-I and -III) in the brain of the nonparasitic species, the American brook lamprey, Lampetra appendix, for comparison with the extensive, published, immunohistochemical data on these hormones in the parasitic species, the sea lamprey, Petromyzon marinus. The complete cDNAs for the two lamprey prohormones, proopiocortin (POC), and proopiomelanotropin (POM), were cloned for L. appendix and both nucleotide and deduced amino acid sequences were compared with those previously published for P. marinus. The POC and POM cDNAs for both species were used in expression studies, with Northern blotting, throughout their life cycles. Although GnRH-I and -III immunohistochemistry revealed a similar distribution of immunoreactive cells and fibers in the two species during the life cycles, a qualitative evaluation of staining intensity in L. appendix, implied early activity in the brains of metamorphosis of this species, particularly in GnRH-I. GnRH-III seems to be important in larval life and early metamorphosis in both species. A novel feature of this immunohistochemical study is the monthly observations of the distribution and relative intensity of the two GnRHs during the critical period of final sexual maturation that lead to spawning and then the spent animal. L. appendix POC and POM nucleotide sequences had 92.9 and 94.6% identity, respectively, with P. marinus POC and POM and there was an earlier increase in their expression during metamorphosis and postmetamorphic life. Since there was some correlation between the timing of metamorphic development, gonad maturation, and brain irGnRH intensity with POC and POM expression in L. appendix, it was concluded that these prohormones yield posttranslational products that likely play a substantial role in development and maturation events that lead to the nonparasitic adult life history of this species. PMID- 16364324 TI - Indications associated with increased cesarean section rates in a Swedish hospital. AB - OBJECTIVE: To analyze the indications and Robson classes associated with the rapid increase in cesarean section (CS) rates at Soder Hospital, Stockholm, Sweden, in the late 1990s. METHOD: Records of women who underwent CSs in 1994 and 1999 at Soder Hospital were retrospectively reviewed. Diagnostic frequency and Robson class, which takes into account characteristics such as parity, previous deliveries, prematurity, and fetal presentation, were compared for the 2 years. RESULTS: Suspected fetal distress (+1.6%; P = .0001), maternal request (+1.5%; P < .0001), and labor dystocia (+0.8%; P = .03) were associated with the increase in CS rates. The rate of CSs with cephalic presentation and spontaneous onset of labor at term, as well as the rate of CSs following induced labor or elective CSs, increases significantly in both nulliparas and multiparas (Robson classes 1 4) (P < .02). CONCLUSION: The increasing CS rate was due to maternal preference and lower thresholds of decision for physicians. PMID- 16364325 TI - Short-term therapy for mixed vaginal infections. PMID- 16364326 TI - Pregnancy outcomes in women with heart disease. PMID- 16364327 TI - Minilaparotomy and chromopertubation as an alternative to laparoscopy. PMID- 16364328 TI - Association between ectopic pregnancy and pelvic endometriosis. PMID- 16364329 TI - Towards a geology of evidence-based practice--a discussion paper. AB - We begin this paper with a consideration of the significance of a historical perspective in presentations of evidence-based practice in the nursing and medical literature. We suggest that whereas writers often produce coherent historical narratives as justification for particular views of the nature of EBP, an examination of its origins reveals no such signs of historical development or progress in our conception or understanding of it. We then explore alternative modes of thought for attempting to understand and critique the variety of definitions and descriptions of EBP to be found in the literature. We eventually reject the linear mode of historical thinking in favour of Deleuze's notion of rhizomatic thought and the metaphor of geology. Finally, we employ the rhizomatic mode of thinking and writing to construct a geology of evidence-based practice which attempts to expose and embrace contradictions in definitions and uses of the term rather than discount them in an authorised historical narrative written from the perspective of the dominant discourse. PMID- 16364330 TI - Muscle fascicle and series elastic element length changes along the length of the human gastrocnemius during walking and running. AB - Ultrasound imaging has recently been used to distinguish the length changes of muscle fascicles from those of the whole muscle tendon complex during real life movements. The complicated three-dimensional architecture of pennate muscles can however cause heterogeneity in the length changes along the length of a muscle. Here we use ultrasonography to examine muscle fascicle length and pennation angle changes at proximal, distal and midbelly sites of the human gastrocnemius medialis (GM) muscle during walking (4.5 km/h) and running (7.5 km/h) on a treadmill. The results of this study have shown that muscle fascicles perform the same actions along the length of the human GM muscle during locomotion. However the distal fascicles tend to shorten more and act at greater pennation angles than the more proximal fascicles. Muscle fascicles acted relatively isometrically during the stance phase during walking, however during running the fascicles shortened throughout the stance phase, which corresponded to an increase in the strain of the series elastic elements (SEEs) (consisting of the Achilles tendon and aponeurosis). Measurement of the fascicle length changes at the midbelly level provided a good approximation of the average fascicle length changes across the length of the muscle. The compliance of the SEE allows the muscle fascicles to shorten at a much slower speed, more concomitant with their optimal speed for maximal power output and efficiency, with high velocity shortening during take off in both walking and running achieved by recoil of the SEE. PMID- 16364331 TI - Comparison of the finite helical axis and the rectangular coordinate system in representing orthodontic tooth movement. AB - In orthodontics, tooth movement is typically described using the rectangular coordinate system (XYZ); however, this system has several disadvantages when performing biomechanical analyses. An alternative method is the finite helical axis (FHA) system, which describes movement as a rotation about and a translation along a single axis located in space. The purpose of this study was to examine differences between the FHA and the XYZ systems in analyzing orthodontic tooth movement. Maxillary canine retraction was done using sliding mechanics or a retraction spring with midpalatal orthodontic implants used as measuring references. Tooth movement calculated with the FHA was compared with the corresponding movement in the rectangular coordinate system weekly over a 2-month interval in eight patients. The FHA showed that sliding mechanics controlled rotation of the canine better than the retraction spring (Ricketts retractor), and that the Ricketts retractor controlled tipping better. Changes in the FHA direction and position vectors with time showed that the biomechanical forces are not uniform during the treatment period. In both mechanics, the FHA provided a simple biomechanical model for canine retraction. PMID- 16364332 TI - Dicarboxylic degradation products of nonylphenol polyethoxylates. Determination and structural elucidation in water samples by solid-phase extraction and gas chromatography-mass spectrometry after methylation. AB - A reliable method combining solid-phase extraction, derivatization and gas chromatography-chemical ionization mass spectrometry (GC-CI-MS) was developed for the measurement, in river and sewage effluent water, of four select model compounds of dicarboxylic metabolites (dm-CA(5-8)P1EC) and other dicarboxylic metabolites (CA(5-8)P1ECs) of nonylphenol polyethoxylates. These selected isomers were referred as dm-CA(5-8)P1ECs because they have an alpha,alpha-dimethyl configuration (expressed as "dm"), five to eight C atoms and a carboxyl group in the alkyl chain, and an ethoxy acetic acid group. The derivatization of terminal carboxyl groups was successful with (trimethylsilyl)diazomethane. The best extraction conditions were obtained using an Oasis HLB cartridge as a sorbent bed and 4 ml of MTBE/methanol (9:1, v/v) elution mixture. The method detection limits of 0.03-0.07 microg/l for dm-CA(5-8)P1ECs were attained in 500 ml pure water. The recovery was then evaluated for pure water, river and sewage effluent water samples. The high recoveries of typically >89% for each isomer indicated the high performance of the method. Although dm-CA(5-8)P1ECs were not detected in the collected water samples, 21 isomers of CA(5-8)P1ECs were identified by CI-MS and the tentative structures of six out of them were elucidated, mainly limited to the branch at alpha-C atom, by studying the EI-mass spectra. The relative concentrations of individual CA(5-8)P1EC metabolites were calculated based on dm CA(5-8)P1ECs. The results showed that the main degradation on the nonyl chain occurred via the elimination of two carbon-units and the concentrations in Japan were much lower than those in Taiwan and Italy. PMID- 16364333 TI - Determination of steroids in human plasma using temperature-dependent inclusion chromatography for metabolomic investigations. AB - Clinical and metabolomic investigations of complex human fluids require cost effective methodologies that can rapidly assess the steroid hormone milieu of individual samples. The efficiency of quantification of many steroids is limited using immunoassays as these methods can only measure a single component of biological samples and are dependent upon the specificity of the antiserum used in the protocol. In this study, we optimised the solid-phase extraction protocol for the extraction of a range of steroids of varied polarity from estetrol to progesterone from human plasma. The final SPE procedure for efficient extraction of steroids was a washing mixture of 5 ml of 30% methanol and an elution solvent of 2 ml of 100% methanol using 0.5 g C-18 cartridges. This protocol resulted in a high recovery rate, ranging from 85.2 to 99.9% for both the internal standard (7,8-dimethoxyflavone) and steroids of interest. We also improved the separation methodology of our previous work using temperature dependent inclusion chromatography with a mobile phase composition of 35% acetonitrile and 12 mM of beta-cyclodextrin at 29 degrees C. Under these conditions most of the fluid components including estetrol were detected in the first 10 min with progesterone appearing at 43 min. This method is simplistic, inexpensive and reproducible with the capabilities of accurate quantification of steroids. Therefore it could have numerous clinical and metabolomic applications. PMID- 16364334 TI - Chemometric treatment of vanillin fingerprint chromatograms. Effect of different signal alignments on principal component analysis plots. AB - This study describes the chemometric treatment of vanillin fingerprint chromatograms to distinguish vanillin from different sources. Prior to principal component analysis, which is used to discriminate vanillin from different origins, the fingerprints are aligned. Three alignment algorithms are tested, correlation optimized warping (COW), target peak alignment (TPA) and semi parametric time warping (STW). The performance of the three algorithms is evaluated and the effect of the different alignments on the PCA score plots is investigated. The alignment obtained with STW differs somewhat from that with COW and TPA. However, equivalent score plots were obtained regarding the different vanillin groups. PMID- 16364335 TI - Enantioseparation of 2-aryloxypropionic acids on chiral porous monolithic columns by capillary electrochromatography. Evaluation of column performance and enantioselectivity. AB - The enantioseparation of 2-aryloxypropionic acids by capillary electrochromatography was tested on columns with a monolithic stationary phase prepared from silanized fused-silica capillaries (100 microm I.D.) by in situ copolymerization of glycidyl methacrylate, ethylene glycol dimethacrylate and methyl methacrylate in the presence of formamide and 1-propanol as the porogen solvents. The porous chiral monolithic stationary phases were prepared by reaction of the epoxy-groups at the surface of the monolith with (+)-1-(4 aminobutyl)-(5R,8S,10R)-terguride. To attain the minimum HETP values for the enantiodiscrimination of 2-phenoxypropionic acid, the influence of the composition of polymerization solution on column total porosity and efficiency was investigated. Optimum mobile phase conditions were found for all analytes tested using acetonitrile-methanol mixtures containing triethylamine and acetic acid as the buffer components. Furthermore, the chemical and mechanical stabilities of the columns were satisfactory, allowing hundreds of analyses. PMID- 16364336 TI - Development of analytical methods for multiresidue determination of quinolones in pig muscle samples by liquid chromatography with ultraviolet detection, liquid chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry. AB - This paper presents a comparison between liquid chromatography with ultraviolet detection (LC-UV), liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods developed for the multiresidue determination of 8 quinolones, around their maximum residue levels (MRLs) in pig muscle. The procedure involves common extraction of the quinolones from the tissues by traditional extraction, a step for clean-up and preconcentration of the analytes by solid-phase extraction (SPE) and a subsequent liquid chromatographic analysis. The methods present satisfactory results of linearity, precision and limits of quantification much lower than the MRLs established by the European Union for quinolones in pig tissues. PMID- 16364337 TI - Structural transition of glucagon in the concentrated solution observed by electrophoretic and spectroscopic techniques. AB - Glucagon, a polypeptide hormone consisting of 29 amino acid residues, tends to form gel-like fibrillar aggregates, and the glucagon fibril, as well as other pathologically related fibrils including prion, amylin, and beta-amyloid, have been found to be cytotoxic through the activation of apoptotic signaling pathways. To understand the aggregation properties of glucagon fibril, we have characterized and compared the physicochemical properties of glucagon, secretin, a member of the glucagon superfamily, and amylin using analytical techniques including capillary electrophoresis (CE), circular dichroism (CD), FT-IR, FT Raman, transmission electron microscopy (TEM), and beta-sheet-imaging probe. Aging treatment of glucagon resulted in the formation of fibrillar aggregates in time- and concentration-dependent manner, and FT-IR and FT-Raman analyses showed the spectral shift of amide I band, suggesting the conformational changes from alpha-helix to beta-sheet structure. Interestingly, secretin, having high sequential and secondary structural homology with glucagon, did not generate the fibrillar aggregates at the conditions tested. In addition, we evaluated the association state of glucagon at various pHs raging from pH 2.0 to 3.5 using CE. Based on the CE data, the rate constants of glucagon aggregation were calculated to be 0.002 +/- 0.004/h and 0.080 +/- 0.011/h for aging at pH 2.0 and 3.5, respectively, suggesting the pH dependence of self-association. CE showed the potential to separate and detect the glucagon aggregates and intermediates during aging process. PMID- 16364339 TI - Performance limits of monolithic and packed capillary columns in high-performance liquid chromatography and capillary electrochromatography. AB - A method is proposed for the comprehensive characterization and comparison of columns in the high-performance liquid chromatographic (HPLC) and capillary electrochromatographic (CEC) modes. Using this approach, column parameters such as the number of plates, the eddy-diffusion and mass-transfer contributions to peak broadening, the permeability, and the analysis time are incorporated in a single graph and a comparison in terms of efficiency and speed is obtained. The chromatographic performance of silica-based and polymer-based monolithic capillary columns is discussed and a comparison is made with the performance of packed columns. Also, the potential of ultra-high-pressure liquid chromatography is discussed in this context. In the HPLC mode, the best results were obtained with silica monoliths; in the CEC mode, the low-density methacrylate-ester-based monoliths showed the best performance. PMID- 16364338 TI - Analysis of frankincense from various Boswellia species with inhibitory activity on human drug metabolising cytochrome P450 enzymes using liquid chromatography mass spectrometry after automated on-line extraction. AB - In our search for herbal remedies with inhibitory activity on cytochrome P450 (CYP) enzymes, we identified extracts of the gum-resin of Boswellia carteri, Boswellia frereana, Boswellia sacra and Boswellia serrata as equally potent, non selective inhibitors of the major drug metabolising CYP enzymes 1A2/2C8/2C9/2C19/2D6 and 3A4. LC/LC/ESI-MS fingerprint analyses of the boswellic acids 11-keto-beta-boswellic acid, alpha-boswellic acid, beta-boswellic acid and their 3-O-acylated derivatives were used for the authentication of the commercially obtained frankincense samples. Although the boswellic acids could be identified as moderate to potent inhibitors of the applied CYP enzymes, they are not the major CYP inhibitory principle of frankincense. PMID- 16364340 TI - Development of an analytical method using microchip capillary electrophoresis for the measurement of fluorescein-labeled salivary components in response to exercise stress. AB - We have developed an analytical method using microchip capillary electrophoresis (microchip CE) for the high-speed separation of fluorescein-labeled salivary components in response to exercise stress. Optimal separation was obtained using a borate buffer at pH 9.5 containing 10 mM beta-cyclodextrin and 1.0% (w/v) methylcellulose. To minimize individual differences in human saliva, such as viscosity, conductivity, and contaminants, the concentration of methylcellulose in the analytical conditions played a key factor. The optimized separation conditions produced identical electropherograms successfully despite of the use of different microchips made from quartz glass or poly-methylmethacrylate (PMMA). In addition, a practical application of bicycle ergometer stress was performed. Some components in human saliva showed a marked decrease after exercise stress. PMID- 16364341 TI - Preparative isolation and purification of four compounds from the Chinese medicinal herb rhizoma Anemarrhenae by high-speed counter-current chromatography. AB - High-speed counter-current chromatography (HSCCC) was applied to the separation and purification of mangiferin, neomangiferin, cis-hinkiresinol and (-)-4'-O methylnyasol from the Chinese medicinal herb rhizoma Anemarrhenae. Five hundred milligrams of crude extracts were separated by using n-butanol-acetic acid (1%) (1:1, v/v) as the two-phase solvent system and yielded 35.3 mg of neomangiferin and 245.4 mg of mangiferin. During this separation, cis-hinkiresinol and (-)-4'-O methylnyasol were still maintained in the stationary phase. The stationary phase was collected, evaporated to dryness and separated with light petroleum-ethyl acetate-methanol-water (1:1:1.2:0.8, v/v) and 1:1:1.4:0.6 (v/v) in gradient elution, which yielded 17.2 mg of cis-hinkiresinol and 12.4 mg of (-)-4'-O methylnyasol. The purities of mangiferin, neomangiferin, cis-hinkiresinol and (-) 4'-O-methylnyasol were 96.3, 98.0, 97.3 and 98.2%, respectively, as determined by HPLC. The chemical structures of these components were identified by 1H NMR and 13C NMR. PMID- 16364342 TI - Preparative separation of cichoric acid from Echinacea purpurea by pH-zone refining counter-current chromatography. AB - pH-zone-refining counter-current chromatography was successfully applied to the separation of cichoric acid from Echinacea Purpurea (L.) Moench. A 3.0 g quantity of sample was separated using the following two-phase solvent system: MtBE-CH3CN water (4:1:5, v/v), 10 mM trifluoroacetic acid in organic stationary phase and 10 mM ammonia in aqueous mobile phase. The obtained fractions were analyzed by high performance liquid chromatography and negative ion electrospray ionization mass spectrometry. Double separations were performed with the same solvent system yielding 563 mg cichoric acid at 95.6% purity. PMID- 16364343 TI - High-throughput analysis of bergamot essential oil by fast solid-phase microextraction-capillary gas chromatography-flame ionization detection. AB - The advantages of using a narrow-bore column in headspace solid-phase microextraction-gas chromatographic (HS-SPME-GC) analysis are investigated. An automated rapid HS-SPME-GC method for the determination of volatile compounds in a complex sample (bergamot essential oil) was developed. A low-capacity (7 microm) SPME fibre was employed, enabling a short equilibration time (15 min). The absorbed volatile compounds were then separated in 12.5 min on a 10 m x 0.1 mm I.D. capillary. The fast GC method was characterized by relatively moderate GC parameters (head pressure: 173 kPa; temperature program rate: 12 degrees C/min). The employment of the low-capacity fibre also suited the reduced sample capacity of the capillary employed, hence column overloading was avoided. Analytical repeatibility was determined in terms of retention times (maximum RSD: 0.32%) and peak areas (maximum RSD: 9.80%). The results obtained were compared to those derived from a conventional HS-SPME-GC (a 30 microm SPME fibre and 0.25 mm I.D. capillary were used) application on the same sample. In this respect, a great reduction of analytical time was obtained both with regard to the conventional SPME equilibration and GC run times, which both required 50 min. Peak resolution was altogether comparable in both applications. Although a slight loss in terms of sensitivity was observed in the rapid approach (generally within the 25-50% range), this did not impair the detection of all peaks of interest. Finally, the selectivities of the 30 and 7 microm fibres were evaluated and, as expected, these were in good agreement. PMID- 16364344 TI - Retention behavior of silica-bonded and novel polymeric reversed-phase sorbents in studies on flavones as chemotaxonomic markers of Scutellaria L. genus. AB - In this study, the effectiveness of both classical and novel polymeric sorbents used in solid-phase extraction (SPE) for isolation of pharmacologically active flavones (baicalin, luteolin, apigenin, wogonin and chrysin) from aerial parts of 13 species of Scutellaria L. (Skullcap) genus was assessed. The highest recoveries of hydrophilic (baicalin, luteolin) flavones for Oasis HLB were obtained, whereas for medium hydrophobic (apigenin) and hydrophobic (wogonin, chrysin) flavones better quantitative results for BakerBond phenyl cartridges were stated. Eluates were analysed using reversed-phase high-performance liquid chromatography with diode array detection (RP-HPLC-DAD). For the five target compounds the determined concentrations ranged from approximately 4 to approximately 15,500 microg/g dry wt. Very good linearities (r(2)>0.9995) of calibration curves were achieved for each flavone. The accuracy was below 5% for most compounds examined. This is the first method reported that enabled simultaneous qualification and quantitation of five flavones (being chemotaxonomic markers) in 13 species of Scutellaria L. genus. PMID- 16364345 TI - Comparison of analytical and semi-preparative columns for high-performance liquid chromatography-solid-phase extraction-nuclear magnetic resonance. AB - The application of analytical and semi-preparative columns in reversed-phase liquid chromatography-solid-phase extraction-nuclear magnetic resonance (HPLC-SPE NMR) was compared. The work was aiming at separating a higher sample amount in a single run and in this way to reduce the necessary NMR measurement time of separated compounds. Several parameters for compound separation and trapping procedures were optimised: flow rate of HPLC and make-up water pumps, choice of stationary phase cartridges and drying time. The separation and loadability of nine model compounds on analytical and semi-preparative columns was determined, as well as the focussing capacity of SH-type SPE cartridges. It was found that a semi-preparative column--or multiple peak trapping on analytical columns--gave better results than a standard 4.6mm analytical column for non-polar compounds (e.g. flavonoid aglycones, sesquiterpene lactones, non-polar terpenes, logP>2), but for polar compounds (logP<-2) did not offer any advantage over an analytical column, or was even disadvantageous. For intermediately polar compounds ( 2 0.99) over a concentration range of 0.16-0.80 microg/spot. Recovery from the HMP capsules is statistically equal to 100%. The precision of the method with respect to time and concentration is acceptable, with relative standard deviation (RSD) values of 1.28 and 1.49%, respectively. PMID- 16364348 TI - Tandem column for the simultaneous determination of arginine, ibuprofen and related impurities by liquid chromatography. AB - Ibuprofen arginate is a rapidly absorbed salt designed to promote more rapid onset of analgesia than commercially available forms of ibuprofen. Ibuprofen and arginine have very different polarities and this becomes in a chromatographic problem, further complicated with the determination of related compounds, which is necessary in stability assays of the pharmaceutical forms. The common solution is the employment of two separate methods, but this is time consuming. A LC method has been developed to determinate both compounds and related impurities in one run. Ibuprofen, arginine and three ibuprofen related impurities (B, E and J) have been baseline separated with isocratic conditions at pH 3.0 and run time under 20 min by employing a tandem combination of two different stationary phases: first a ZORBAX SB-C18 column from Agilent (250 mm x 4.6 mm and 5 microm) and downstream a SUPELCOSIL LC-NH2 column from Supelco (150 mm x 4.6 mm and 3 microm). The octadecyldiisobutylsilane column provides the separation of ibuprofen and its impurities by a hydrophobic mechanism, whereas aminopropyl column offers selective retention of arginine by dipolar interaction mechanism. Method has been successfully validated following ICH guidelines and it has been demonstrated to be reliable for arginine, ibuprofen and related impurities determination in sachets of two different dosages as pharmaceutical forms. Moreover, stress test has proved the selectivity of the method for degradation products, such as those that can emerge throughout long-term stability assays. PMID- 16364349 TI - N-linked oligosaccharide analysis of rat brain Thy-1 by liquid chromatography with graphitized carbon column/ion trap-Fourier transform ion cyclotron resonance mass spectrometry in positive and negative ion modes. AB - We have previously described the site-specific glycosylation analysis of rat brain Thy-1 by LC/multistage tandem mass spectrometry (MS(n)) using proteinase digested Thy-1. In the present study, detailed structures of oligosaccharides released from Thy-1 were elucidated by mass spectrometric oligosaccharide profiling using LC/MS with a graphitized carbon column (GCC-LC/MS). First, using model oligosaccharides, we improved the oligosaccharide profiling by ion trap mass spectrometry (IT-MS) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Sequential scanning of a full MS(1) scan with FT ICR-MS followed by data-dependent MS(n) with IT-MS in positive ion mode, and a subsequent full MS(1) scan with FT-ICR-MS followed by data-dependent MS(n) with IT-MS in negative ion mode enabled the monosaccharide composition analysis as well as profiling and sequencing of both neutral and acidic oligosaccharides in a single analysis. The improved oligosaccharide profiling was applied to elucidation of N-linked oligosaccharides from Thy-1 isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was demonstrated that Thy-1 possesses a significant variety of N-linked oligosaccharides, including Lewis a/x, Lewis b/y, and disialylated structure as a partial structure. Our method could be applicable to analysis of a small abundance of glycoproteins, and could become a powerful tool for glycoproteomics. PMID- 16364350 TI - Simultaneous determination of synthetic phosphodiesterase-5 inhibitors found in a dietary supplement and pre-mixed bulk powders for dietary supplements using high performance liquid chromatography with diode array detection and liquid chromatography-electrospray ionization tandem mass spectrometry. AB - A high-performance liquid chromatography-diode array detection (HPLC-DAD) method and a liquid chromatography-electrospray ionization tandem mass spectrometry (LC ESI-MS/MS) method were developed to screen for the presence of synthetic phosphodiesterase-5 (PDE-5) inhibitors and their analogues, namely sildenafil, vardenafil, tadalafil, homosildenafil, acetildenafil and hydroxyhomosildenafil. The methods were applied to pre-market samples submitted to the Health Sciences Authority of Singapore (HSA) for testing. One sample was in the form of capsules while six other samples were pre-mixed bulk powder samples for dietary supplements to be repackaged or formulated into the final dosage forms (usually capsules). Identification of PDE-5 inhibitors and their analogues was achieved by comparing individual peak retention times, UV spectra and mass spectra with those of reference standards. The seven samples were found to contain at least one of the following compounds: sildenafil, vardenafil, hydroxyhomosildenafil, homosildenafil and acetildenafil. The five compounds were simultaneously determined by LC-ESI-MS/MS in multiple reactions monitoring (MRM) scan mode. The method has been validated for accuracy, precision, linearity and sensitivity. PMID- 16364351 TI - Porous structure and fluid partitioning in polyethylene cores from 3D X-ray microtomographic imaging. AB - Using oil-wet polyethylene core models, we present the development of robust throat finding techniques for the extraction, from X-ray microtomographic images, of a pore network description of porous media having porosity up to 50%. Measurements of volume, surface area, shape factor, and principal diameters are extracted for pores and area, shape factor and principal diameters for throats. We also present results on the partitioning of wetting and non-wetting phases in the pore space at fixed volume increments of the injected fluid during a complete cycle of drainage and imbibition. We compare these results with fixed fractional flow injection, where wetting and non-wetting phase are simultaneously injected at fixed volume ratio. Finally we demonstrate the ability to differentiate three fluid phases (oil, water, air) in the pore space. PMID- 16364352 TI - Lamellar nanocomposites based on exfoliated SbP2O8- nanosheets and ionic polyacetylenes. AB - Novel lamellar nanocomposites composed of exfoliated SbP2O8- nanosheets and poly(N-methyl-2-ethynyl pyridinium iodide) (PNMe) or poly(N-octadecyl-2-ethynyl pyridinium bromide) (PNO) have been synthesized by exfoliation and re-stacking method. The XRD data of the nanocomposites SbP2O8-:PNMe and PNO:SbP2O8- indicate the formation of nanostructures containing SbP2O8- nanosheets and PNMe or PNO with interlayer expansions of 2.22 and 4.2 nm along the stacking direction of SbP2O8- nanosheets. Formation of these nanocomposites is further supported by the results obtained by FT-IR spectroscopy, scanning transmission electron microscopy and thermogravimetry. The values of electrical conductivity of the polymers and nanocomposites have also been measured. PMID- 16364353 TI - Characterization and adsorption properties of diatomaceous earth modified by hydrofluoric acid etching. AB - This work was a study of the chemical modification of diatomaceous earth (DE) using hydrofluoric acid (HF) solution. Under the experimental conditions investigated, it was found that HF under controlled conditions significantly etched inward into the interior of the existing pore structure in the clay mineral due to its high content of silica, leaving a framework possessing a larger BET surface area (ca. 10 m2 g(-1)) in comparison with that (ca. 4 m2 g( 1)) of its precursor (i.e., DE). Further, the results indicated that the HF concentration is a more determining factor in creating more open pores than other process parameters (temperature, holding time, and solid/liquid ratio). This observation was also in close agreement with the examinations by the silicon analysis, scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The adsorption kinetics and the adsorption isotherm of methylene blue onto the resulting clay adsorbent can be well described by a pseudo-second-order reaction model and the Freundlich model, respectively. PMID- 16364354 TI - Chemical potential of a hard sphere fluid adsorbed in model disordered polydisperse matrices. AB - We consider a model for adsorption of a simple fluid in disordered polydisperse adsorbents. The fluid consists of hard sphere particles. On the other hand, the adsorbents of this study are modeled as a collection of hard spheres with their diameter obeying a certain distribution function. Our focus is in the evaluation of the chemical potential of the fluid immersed in such a polydisperse material. It permits us to obtain porosity and pore size distribution for the adsorbent, as well as a set of adsorption isotherms. The latter have been calculated theoretically and by grand canonical Monte Carlo simulations. We observe that the width of assumed polydispersity distribution affects all the properties of the system. Nevertheless, the effect of matrix packing is dominant in determining adsorption for this class of models. We are convinced that the matrix structures generated via more sophisticated algorithms would exhibit stronger effects of polydispersity on the entire set of properties of adsorbed simple fluids. PMID- 16364355 TI - Self-assembly in evaporated polymer solutions: influence of the solution concentration. AB - Amorphous polymers were dissolved in chlorinated organic solvents and deposited on thin horizontal substrates. The solutions with various concentrations of polymers were deposited and evaporated under ambient conditions in a slow air current. Self-assembled oriented mesoscopically scaled patterns were observed. The patterns were studied with optical and atomic force microscopy. The concentration of the solution exerts a decisive influence on the mesoscopic cell characteristic dimension. Cell dimensions grow linearly with the polymer solution concentration for all kinds of tested polymers, chlorinated solvents and substrates. The dependence could be explained by a physical mechanism, based on the mass transport instability occurring under the intensive evaporation of the solvent. In situ FTIR study of the process was performed first. It was established with FTIR spectroscopy that film porosity is not due to water droplet condensation under evaporation. PMID- 16364356 TI - Breakup of carbon nanotube flocs in microfluidic traps. AB - The critical stress to break flocs of multiwalled-carbon nanotubes suspended in low-molecular-weight polymer fluid is measured in planar elongational flow. Through image analysis of aggregates and their fragments, the extension rate of the flow and the size and aspect ratio of the aggregates are measured in real time. While trapping an aggregate at the stagnation point of the device, the flow rate is increased continually and breaking events are recorded, establishing a correlation between aggregate size and the critical elongation stress for fragmentation sigma. This relationship resembles that for the breakup of flocs comprising spherical particles, yet it indicates that the strength of these fibrous flocs depends much more strongly on the local particle concentration. Fracture avalanches (or cascades) are also observed. PMID- 16364357 TI - Diffusioosmosis of electrolyte solutions in a fine capillary slit. AB - The steady diffusioosmotic flows of an electrolyte solution along a charged plane wall and in a capillary channel between two identical parallel charged plates generated by an imposed tangential concentration gradient are theoretically investigated. The plane walls may have either a constant surface potential or a constant surface charge density. The electrical double layers adjacent to the charged walls may have an arbitrary thickness and their electrostatic potential distributions are determined by the Poisson-Boltzmann equation. Solving a modified Navier-Stokes equation with the constraint of no net electric current arising from the cocurrent diffusion, electric migration, and diffusioosmotic convection of the electrolyte ions, the macroscopic electric field and the fluid velocity along the tangential direction induced by the imposed electrolyte concentration gradient are obtained semianalytically as a function of the lateral position in a self-consistent way. The direction of the diffusioosmotic flow relative to the concentration gradient is determined by the combination of the zeta potential (or surface charge density) of the wall, the properties of the electrolyte solution, and other relevant factors. For a given concentration gradient of an electrolyte along a plane wall, the magnitude of fluid velocity at a position in general increases with an increase in its electrokinetic distance from the wall, but there are exceptions. The effect of the lateral distribution of the induced tangential electric field and the relaxation effect in the double layer on the diffusioosmotic flow are found to be very significant. PMID- 16364358 TI - Effect of CuCl2 concentration on the aggregation and mineralization of Tobacco mosaic virus biotemplate. AB - Aggregation of the biotemplates in mineralization processes is a considerable obstacle in preparing well-dispersed bio-inorganic hybrid materials. In this study, aggregation and mineralization of Tobacco mosaic virus (TMV) biotemplates were investigated as a function of the copper precursor (CuCl2) concentration. The mean hydrodynamic radius of TMV in an aqueous CuCl2 solution was determined by dynamic light scattering for the monitoring of the TMV aggregation. At CuCl2 concentration of 0.5 mM or higher, the mean hydrodynamic radius of TMV increased dramatically indicating aggregation of the TMV particles. Numerical calculations on the long-range interaction energy between parallel model TMV particles agreed with the experimental observations for the TMV aggregation. Mineralization of copper precursors on the TMV biotemplates was achieved only at the CuCl2 concentrations that induced considerable aggregation of the biotemplate. From the numerical calculations and experimental results, it was concluded that a dense copper cluster deposition cannot be achieved without aggregation of TMV templates. PMID- 16364359 TI - Influence of interfacial features in the supramolecular assembly of 3,5 dihydroxybenzylalcohol based dendrimers on Si/SiO2 surfaces. AB - Self-organization of 3,5-dihydroxybenzylalcohol (DHBA) based dendrimers of generations 0-3 (G0-G3) on bare and functionalized single crystal silicon (Si/SiO2) surfaces has been examined. The underlying monolayer plays a significant role in the supramolecular assembly leading to ordered structures of DHBA (G0) and generation 1-3 (G1-G3) dendrimers at interfaces. Ordered hyperbranched structures are formed on surfaces containing self-assembled monolayers with complimentary features to the assembling molecules, whereas no such organized assemblies are observed on unfunctionalized surfaces. PMID- 16364360 TI - Children use vowels to help them spell consonants. AB - English spelling is highly inconsistent in terms of simple sound-to-spelling correspondences but is more consistent when context is taken into account. For example, the choice between ch and tch is determined by the preceding vowel (coach, roach vs. catch, hatch). We investigated children's sensitivity to vowel context when spelling consonants in monosyllabic nonwords. Second graders (7-year olds) tended to use vowel context correctly when spelling word-final consonants (codas). This use of context was progressively stronger for third and fifth graders as well as for college students. The increase is not due to differences in vocabulary because the contextual patterns are similar in reading materials targeted at all four age groups. Vowel letters (graphotactics) had a stronger influence than did vowel pronunciation. Children also used vowel context when spelling word-initial consonants (onsets); this effect was as strong for second graders as for adults. Thus, novice spellers take advantage of graphotactic information. PMID- 16364361 TI - The solvation interface is a determining factor in peptide conformational preferences. AB - The 21 residue polyalanine-based F(s) peptide was studied using thousands of long, explicit solvent, atomistic molecular dynamics simulations that reached equilibrium at the ensemble level. Peptide conformational preference as a function of hydrophobicity was examined using a spectrum of explicit solvent models, and the peptide length-dependence of the hydrophilic and hydrophobic components of solvent-accessible surface area for several ideal conformational types was considered. Our results demonstrate how the character of the solvation interface induces several conformational preferences, including a decrease in mean helical content with increased hydrophilicity, which occurs predominantly through reduced nucleation tendency and, to a lesser extent, destabilization of helical propagation. Interestingly, an opposing effect occurs through increased propensity for 3(10)-helix conformations, as well as increased polyproline structure. Our observations provide a framework for understanding previous reports of conformational preferences in polyalanine-based peptides including (i) terminal 3(10)-helix prominence, (ii) low pi-helix propensity, (iii) increased polyproline conformations in short and unfolded peptides, and (iv) membrane helix stability in the presence and absence of water. These observations provide physical insight into the role of water in peptide conformational equilibria at the atomic level, and expand our view of the complexity of even the most "simple" of biopolymers. Whereas previous studies have focused predominantly on hydrophobic effects with respect to tertiary structure, this work highlights the need for consideration of such effects at the secondary structural level. PMID- 16364362 TI - Insights on a new PDI-like family: structural and functional analysis of a protein disulfide oxidoreductase from the bacterium Aquifex aeolicus. AB - A potential role in disulfide bond formation in the intracellular proteins of thermophilic organisms has recently been attributed to a new family of protein disulfide isomerase (PDI)-like proteins. Members of this family are characterized by a molecular mass of about 26kDa and by two Trx folds, each comprising a CXXC active site motif. We report on the functional and structural characterization of a new member of this family, which was isolated from the thermophilic bacterium Aquifex aeolicus (AaPDO). Functional studies have revealed the high catalytic efficiency of this enzyme in reducing, oxidizing and isomerizing disulfide bridges. Site-directed mutagenesis experiments have suggested that its two active sites have similar functional properties, i.e. that each of them imparts partial activity to the enzyme. This similarity was confirmed by the analysis of the enzyme crystal structure, which points to similar geometrical parameters and solvent accessibilities for the two active sites. The results demonstrated that AaPDO is the most PDI-like of all prokaryotic proteins so far known. Thus, further experimental studies on this enzyme are likely to provide important information on the eukaryotic homologue. PMID- 16364363 TI - Involvement of polynucleotide kinase in a poly(ADP-ribose) polymerase-1-dependent DNA double-strand breaks rejoining pathway. AB - Efficient DNA double-strand break (DSB) repair is critical for the maintenance of genomic integrity. In mammalian cells, DSBs are preferentially repaired by the non-homologous end-joining pathway relying on DNA-PK activity, but other mechanisms may promote end-joining. We previously described a DSB repair pathway that requires synapsis of DNA ends by poly(ADP-ribose) polymerase-1 (PARP-1) and ligation by the XRCC1/DNA ligase III complex (XL). Here, the repair of non ligatable DNA ends by this pathway was examined in human cell extracts. The phosphorylation of the 5'-terminal end was shown to represent a limiting step for the repair process. Polynucleotide kinase (hPNK) was identified as the 5'-DNA kinase associated with the PARP-1-dependent end-joining pathway because (i) hPNK was co-recruited to DNA ends together with PARP-1 and XL, (ii) ligation of 5'-OH terminal breaks was compromised in hPNK-depleted extracts and restored upon addition of recombinant hPNK, and (iii) recombinant hPNK was necessary for end joining of 5'-OH terminal breaks reconstituted with the PARP-1/XL complex. Also, using an assay enabling us to follow the ligation kinetics of each strand of a DSB, we established that the two strands at the junction can be processed and joined independently, so that one strand can be ligated without a ligatable nick on the other strand at the DSB site. Taken together these results reveal functional parallels between the PARP-1 and DNA-PK-dependent end-joining processes. PMID- 16364364 TI - Equilibrium phi-analysis of a molten globule: the 1-149 apoflavodoxin fragment. AB - The apoflavodoxin fragment comprising residues 1-149 that can be obtained by chemical cleavage of the C-terminal alpha-helix of the full-length protein is known to populate a molten globule conformation that displays a cooperative behaviour and experiences two-state urea and thermal denaturation. Here, we have used a recombinant form of this fragment to investigate molten globule energetics and to derive structural information by equilibrium Phi-analysis. We have characterized 15 mutant fragments designed to probe the persistence of native interactions in the molten globule and compared their conformational stability to that of the equivalent full-length apoflavodoxin mutants. According to our data, most of the mutations analysed modify the stability of the molten globule fragment following the trend observed when the same mutations are implemented in the full-length protein. However, the changes in stability observed in the molten globule are much smaller and the Phi-values calculated are (with a single exception) below 0.4. This is consistent with an overall and significant debilitation of the native structure. Nevertheless, the fact that the molten globule fragment can be stabilised using as a guide the native structure of the full-length protein (by increasing helix propensity, optimising charge interactions and filling small cavities) suggests that the overall structure of the molten globule is still quite close to native, in spite of the lowered stability observed. PMID- 16364365 TI - Probing the mechanism of amyloidogenesis through a tandem repeat of the PI3-SH3 domain suggests a generic model for protein aggregation and fibril formation. AB - Aggregation of the SH3 domain of the PI3 kinase, both as a single domain and as a tandem repeat in which the C terminus of one domain is linked to the N terminus of another by a flexible linker of ten glycine/serine residues, has been studied under a range of conditions in order to investigate the mechanism of protein aggregation and amyloid formation. The tandem repeat was found to form amyloid fibrils much more readily than the single domain under the acidic conditions used here, and the fibrils themselves have higher morphological homogeneity. The folding-unfolding transition of the PI3-SH3 domain shows two-state behaviour and is pH dependent; at pH 3.6, which is near the pH mid-point for folding and only slightly below the isoelectric point of the protein, both the single domain and the tandem repeat spontaneously form broad distributions of soluble oligomers without requirement for nucleation. Under prolonged incubation under these conditions, the oligomers convert into thin, curly fibrils that interact with thioflavin-T, suggesting that they contain an organised beta-sheet structure. Under more acidic conditions (pH 2.0) where the proteins are fully denatured and carry a positive net charge, long, straight fibrils are formed in a process having a pronounced lag phase. The latter was found to be reduced dramatically by the addition of oligomers exceeding a critical size of approximately 20 molecules. The results suggest that the process of aggregation of these SH3 domains can take place by a variety of mechanisms, ranging from downhill formation of relatively amorphous species to nucleated formation of highly organised structures, the relative importance of which varies greatly with solution conditions. Comparison with the behaviour of other amyloidogenic systems suggests that the general mechanistic features outlined here are likely to be common to at least a wide variety of peptides and proteins. PMID- 16364366 TI - Cerebral arteriopathy with extracranial artery involvement in a patient with ulcerative colitis. AB - Arteriopathy of the central nervous system (CNS) complicated with ulcerative colitis is a rare condition, moreover the involvement of extracranial arteries has not been documented. An 18-year-old female complained of a severe pulsatile headache and nausea. She had been diagnosed and treated for ulcerative colitis for four years. Magnetic resonance imaging of the brain showed normal results; however, magnetic resonance angiography (MRA) revealed severe irregularity of the intracerebral arteries. After treatment with prednisolone, the patient fully recovered and the irregularity of the intracerebral arteries was dramatically improved. Vasculitis was strongly suggested as the cause of arteriopathy of the CNS in the present case. Involvement of extracranial arteries such as the carotid artery was also incidentally discovered by duplex ultrasonography and the HLA typing suggested genetic susceptibility to Takayasu's arteritis. Findings from our patient suggest that extracranial arterial involvement should be considered in the case of arteriopathy of the CNS associated with ulcerative colitis. PMID- 16364367 TI - Seed and pollen flow in expanding a species' range. AB - The distinct processes of gene flow via seeds and pollen in hermaphrodite plants provide a biological basis for interpreting their different roles in expanding a species' range. A species' range is primarily expanded through the colonization process by seed dispersal and followed by the joint effects of both seed and pollen flow. Here we examined the effects of seed and pollen flow on shaping a species' distribution in one-dimensional space. Our results demonstrate that pollen flow can enhance range expansion when immigrating genes are adaptive to recipient populations, but can shrink a species' range when immigrating genes are maladaptive. The incompletely purging of maladaptive genes from immigrating pollen grains at the gametophyte stage can reinforce the biological barrier to range expansion. The linkage disequilibria attained by immigrating seeds and pollen grains indirectly amplify the effects of the reaction component and further limit a species' range. The cumulative effect from multiple loci each with a small effect can be substantial on altering a species' range when these genes are maladaptive. These theoretical predictions can help understand the role of pollen flow that is incapable of colonizing new habitats in range expansion. PMID- 16364368 TI - Mixed-mode pattern in Doublefoot mutant mouse limb--Turing reaction-diffusion model on a growing domain during limb development. AB - It has been suggested that the Turing reaction-diffusion model on a growing domain is applicable during limb development, but experimental evidence for this hypothesis has been lacking. In the present study, we found that in Doublefoot mutant mice, which have supernumerary digits due to overexpansion of the limb bud, thin digits exist in the proximal part of the hand or foot, which sometimes become normal abruptly at the distal part. We found that exactly the same behaviour can be reproduced by numerical simulation of the simplest possible Turing reaction-diffusion model on a growing domain. We analytically showed that this pattern is related to the saturation of activator kinetics in the model. Furthermore, we showed that a number of experimentally observed phenomena in this system can be explained within the context of a Turing reaction-diffusion model. Finally, we make some experimentally testable predictions. PMID- 16364369 TI - Neonatal handling induces alteration in progesterone secretion after sexual behavior but not in angiotensin II receptor density in the medial amygdala: implications for reproductive success. AB - Neonatal handling affects the hypothalamus-pituitary-gonadal axis in female rats. Indeed, postnatal handling induces anovulatory estrous cycles and decreases sexual receptiveness. On the other hand, Angiotensin II (Ang II) infused into the medial amygdala (MeA) reduces sexual behavior in male and female rats. Considering this, and that gonadal steroid secretion after copulatory behavior is important for reproductive success, the purpose of the present study was to investigate whether the reduction in sexual receptiveness in neonatally handled female rats is mediated by changes in Ang II receptor density in MeA. Moreover, gonadal steroid secretion after sexual behavior was analyzed. Two groups of female Wistar rats were studied: nonhandled (pups were left undisturbed) and handled (pups were handled for 1 min once a day during the first 10 days of life). Once they were 80-85 days old in the evening of the proestrus day, sexual receptiveness was recorded and after that the animals were killed by decapitation. Trunk blood samples were collected, and plasma estradiol and progesterone were measured by radioimmunoassay. The brains were removed for Ang II receptor autoradiography in MeA. The decreased lordosis quotient in the neonatally handled group was confirmed in the present study. Neonatal handling also reduced the progesterone concentration in the plasma, but did not change the estradiol and the density of Ang II receptors in MeA. The reduced progesterone could be due to the decreased lordosis frequency of handled females. However, this decreased sexual receptiveness is not mediated by changes in Ang II receptors in MeA. PMID- 16364371 TI - Assessing PAH exposure in feral finfish from the Northwest Atlantic. AB - Polycyclic aromatic hydrocarbon (PAH) concentrations were examined in small finfish (<30 cm) represented by capelin, sand lance, American plaice, yellowtail flounder and herring collected opportunistically in various NAFO divisions. Analyses were performed on whole fish and in a portion of the samples; concentrations in internal organs were compared to the rest of the carcass. The effect of pool size, size differences within and between species, lipid content and location were examined to interpret PAH concentrations. Measurements were carried out before the development of the Hibernia oil fields and represent baseline levels for future comparison. Limits in assessing future risk that could be due to discharges of produced water or accidental oil spills are also discussed. Increasing knowledge on the bioaccumulation of PAH, on the production of bile metabolites, the formation of DNA-adducts and of the potential toxic effects associated with PAH will lead to better ecosystem management and protection for future generations. PMID- 16364372 TI - Chronic oil pollution harms Magellanic penguins in the Southwest Atlantic. AB - Petroleum pollution is a problem for seabirds along the Southwest Atlantic coast. Twenty-five groups from Salvador, Brazil (12 degrees 58'S) to San Antonio Oeste, Argentina (40 degrees 43'S) survey or rehabilitate sick or oiled seabirds. Four groups, one each in Brazil and Uruguay, and two in Argentina, kept counts of birds found alive and in need of rehabilitation. An average of 63.7% of the seabirds found were Magellanic penguins (Spheniscus magellanicus), with 3869 reported since 1987. Mainly adult penguins were found in Argentina (1605 of 2102 penguins of known age class) and Uruguay (158 of 197). Juveniles were most common in Brazil (234 of 325). Oil fouling was the most frequent cause of injury or sickness. The number of oiled penguins reported in their wintering range has greatly increased since the early 1990s and is strongly correlated with petroleum exports from Argentina. Our results show that chronic petroleum pollution is a problem for wildlife from Southern Brazil through Northern Argentina, and regulations and enforcement are failing to protect living resources. PMID- 16364373 TI - Nitrate and phosphate supplementation to increase toxin production by the marine dinoflagellate Alexandrium tamarense. AB - Alexandrium tamarense toxins have great value in biotechnology research as well as important in connection with shellfish poisoning. The influence of nitrate or nitrate and phosphate supplementation on cell biomass and toxin content were investigated in batch cultures. When cultures at low nitrate (88.2 microM NaNO(3)) were supplemented with 793.8 microM NaNO(3) at day 10 the cell density and cellular toxin contents were increased by 6-29% and 20-76%, respectively, compared with controls, and maximal values were 43,600 cells/ml (day 38) and 0.91 pg/cell (day 31). Supplementation with nitrate at day 14 or with nitrate and phosphate at day 10/14 to the cultures did not increase the cell density compared with the non-supplemented middle nitrate or high phosphate (108 microM NaH(2)PO(4)) cultures, respectively, but increased the cellular toxin contents by an average of 52%. The results showed that supplementation with nitrate or with nitrate and phosphate at different growth phases of the cultures increased toxin yield by an average of 46%. Supplementation with nitrate at selected times to maintain continuous low level of nitrate might contribute to the effective increase of toxin yield of A. tamarense. PMID- 16364374 TI - Developmental disorders in larvae of eelpout (Zoarces viviparus) from German and Swedish Baltic coastal waters. AB - Pregnant eelpout were collected for several years at German and Swedish Baltic coastal stations with different impact of pollutants and the prevalence of fry abnormalities compared between sites. Mortality of apparently normal developed larvae was observed with high variability in both German and Swedish sites. Malformation of larvae was a prominent disorder at all German stations with a prevalence ranging from about 50% up to a maximum of 90% in almost all analysed samples. At the Swedish stations malformation rates were significantly lower (range: 0-6%). Growth retardation of fry was clearly more prominent in eelpout from German coastal waters as well. Taken together the field data suggests that impaired larval development was associated with the state of environmental pollution at the sampling locations. PMID- 16364375 TI - An alternative approach to modelling relapse in cancer with an application to adenocarcinoma of the prostate. AB - This paper proposes an alternative approach to modelling relapse in cancer. In particular, the dynamic model for the tumor or biomarker will be subjected to a lower elastic boundary at which the process either will be absorbed or reflected. The likelihood of reflection then can be interpreted as the probability of relapse. This framework will be exemplified for prostatic cancer by extending the recently proposed stochastic model of Dayananda et al. [P.W.A. Dayananda, J.T. Kemper, M.M. Shvartsman, A stochastic model for prostate-specific antigen levels, Math. Biosci. 190 (2004) 113] that focussed on the dynamics of the prostate specific antigen (PSA) biomarker. Analytical results for the conditional density function, given a non-negative lower boundary, are obtained for the extreme cases of certain cure and of certain relapse. Simulations illustrate the relevance of the relapse probability and of the normal value of the biomarker for the design of treatment strategies. The paper thus points to two additional (patient specific) characteristics that might enter treatment design and monitoring of progress in therapy. PMID- 16364376 TI - Mutagenesis by man-made mineral fibres in the lung of rats. AB - The potential of two asbestos substitute mineral fibres--rock (stone) wool RW1 and glass wool MMVF10--to induce gene mutations, DNA strand breaks, inflammation and oxidative stress has been studied in rats. Male homozygous lamda-lacI transgenic F344 rats were intratracheally instilled with single doses of 1 and 2 mg/animal of fibres or with multiple doses of 2 mg/animal administered weekly on four consecutive weeks (8 mg in total). Exposure to RW1 fibres for 16 weeks significantly increased mutant frequency (MF) in the lung in a dose-dependent manner, while MMVF10 fibres did not exhibit any increase of MF at any dose. RW1 fibres gave a significant increase of MF at a dose of 1 mg. Four weeks after instillation, neither the single nor the multiple doses significantly increased MF for both fibre types. To investigate mechanisms for induction of mutations, other genotoxicity markers and parameters of inflammatory and oxidative damage were determined in relation to MF. A weak correlation of mutagenicity data with other genotoxicity parameters studied was observed. DNA strand breaks as measured by comet assay were increased in alveolar macrophages and lung epithelial cells of RW1 and MMVF10 treated rats. RWl fibres caused more extensive lung inflammation as measured by release of neutrophils into broncho-alveolar lavage fluid than MMVF10 fibres. The effects were observed 16 weeks post-exposure, indicating a persistence of the pathogenic process during the exposure period. Only minor differences in the extent of inflammatory processes were observed between the doses of 2 mg and 4 x 2 mg, suggesting that any threshold for inflammation lies below the dose of 2 mg. With the exception of the highest dose of MMVF10 fibres after 16 weeks of exposure, no significant increase of oxidative damage as measured by levels of malondialdehyde in lung tissue was observed. MMVF10 fibres caused weaker inflammation in the lung of rats and did not exhibit any mutagenic effect. We conclude that a weak but chronic inflammation (more likely than acute inflammation or direct oxidative damage) in the lung tissue of fibre treated rats characterized by moderate influx of inflammatory cells into BAL is probably responsible for the observed mutagenic effect of RW1 fibres. PMID- 16364378 TI - Visuomotor 'immunity' to perceptual illusion: a mismatch of attentional demands cannot explain the perception-action dissociation. AB - Recent findings of visuomotor immunity to perceptual illusions have been attributed to a perception-action division of labour within two anatomically segregated streams in the visual cortex. However, critics argue that such experimental findings are not valid and have suggested that the perception-action dissociations can be explained away by differential attentional/processing demands, rather than a functional dissociation in the neurologically intact brain: perceptual tasks require processing of the entire illusion display while visuomotor tasks only require processing the target that is acted upon. The present study examined whether grasping of the Muller-Lyer display would remain immune to the illusion when the task required the direction of attention or a related resource towards both Muller-Lyer shafts. Twelve participants were required to match and grasp two Muller-Lyer shafts bimanually (i.e. one with each hand). It was found that bimanual grasping was not significantly affected by the illusion, while there was a highly significant illusion effect on perceptual estimation by matching. Furthermore, it was established that this dissociation did not result from a differing baseline rate of change in manual estimation and grasping aperture to a change in physical object size. These findings provide further support for the postulated perception-action dissociation and fail to uphold the idea that grasping 'immunity' to the Muller-Lyer illusions merely represents an experimental artefact. PMID- 16364377 TI - Chronic administration of dehydroepiandrosterone sulfate (DHEAS) primes for facilitated induction of long-term potentiation via sigma 1 (sigma1) receptor: optical imaging study in rat hippocampal slices. AB - Dehydroepiandrosterone sulfate (DHEAS), one of the most abundant neurosteroids synthesized de novo in the nervous system, has well characterized effects on memory and cognitive performances. However, little is known about the underlying synaptic mechanisms. In this study, we investigated the effects of chronic administration of DHEAS (20 mg/kg for 7 days) on the plasticity of Schaffer collateral-CA1 synapses by applying an optical recording technique on the hippocampal slices stained with voltage-sensitive dyes. We report here that chronically administered DHEAS significantly facilitated the induction of frequency-dependent LTP, termed DHEAS-facilitated LTP. While tetanus of at least 50 pulses (at 100 Hz) were required to induce LTP in control rats, only 20 pulses were needed in DHEAS-treated animals. In contrast DHEA, the non-sulfated form of DHEAS, had no facilitating effect on the induction of LTP. We found that chronically administered DHEAS did not alter the presynaptic glutamate release in response to both single pulse and tetanic stimulation, suggesting that certain alterations happened in postsynaptic neurons. Co-administration of the sigma 1 (sigma1) receptor antagonists, haloperidol or NE100, with DHEAS completely inhibited the DHEAS-facilitated LTP. However, acute administration of sigma1 receptor antagonists to the slices did not affect the induction of DHEAS facilitated LTP, suggesting that sigma1 receptor is a key target of chronic actions of DHEAS but is not involved in the induction of DHEAS-facilitated LTP. Our findings provide evidence that chronically administered DHEAS plays a priming role in inducing a facilitated synaptic plasticity probably via a chronic activation of sigma1 receptor in rat hippocampal CA1 pyramidal cells. PMID- 16364379 TI - Effect of lithium on morphine state-dependent memory of passive avoidance in mice. AB - In the present study, effects of lithium chloride (LiCl) on morphine induced state-dependent memory of passive avoidance task were examined in mice. One-trial step-down paradigm was used for the assessment of memory retention in adult male NMRI mice. Administration of morphine (5 mg/kg) subcutaneously (s.c.) 30 min before training or testing induced impairment of memory performance. Injection of the same dose of the drug 30 min before testing restored memory retention impaired under pre-training morphine effect. Intraperitoneal (i.p.) injection of lithium, 60 min before training or prior to testing also impaired memory performance. Under the pre-training of morphine, the response of the opioid was restored when animals received LiCl (80 and 160 mg/kg) as pre-test injection. Pre training administration of lower dose of lithium (20 mg/kg), but not the higher doses of the drug (80 and 160 mg/kg) impaired memory retention in passive avoidance test. LiCl-induced impairment of memory retention was restored by pre test administration of morphine. In the animals receiving pre-training morphine, combined pre-test morphine and LiCl administration increased the restoration of memory by the opioid. It can be concluded that there may be a cross-state dependency between morphine and lithium. PMID- 16364380 TI - An activity indicator of acute withdrawal depends on amphetamine dose in rats. AB - A moderate dose of amphetamine (AMPH) produces hypoactivity around 20 h post administration. This hypoactivity may be an indicator of an acute withdrawal state. The purpose was to see how AMPH doses affected the expression of this hypoactivity and, by inference, AMPH-induced acute withdrawal. Rats were housed in individual open fields, with free access to food and water. Light-dark cycles were scheduled such that drug-elicited patterns could be readily detected. Animals first received a series of eight control treatments, and then a series of 10 experimental treatments spaced at 33-h intervals. Different experimental treatment groups received saline, 1.0 mg/kg, 2.0 mg/kg, or 4.0 mg/kg AMPH. The effects of these treatments on 33-h patterns of locomotor activity were observed. Control treatments produced no systematic time-dependent changes in activity beyond the first hour post-treatment. All doses of AMPH produced typical short term effects: They markedly increased locomotion and/or stereotypy during the first 3 to 6 h post-treatment. Acute and chronic administrations of the 2.0 and 4.0 mg/kg doses also produced similar changes in longer term activity patterns: They produced hypoactivity 20 h later, followed by a recovery of activity around hour 25 post-treatment. The timing of amphetamine-induced hypoactivity and acute withdrawal may be independent of dose over a wide range of doses. Time-dependent changes in AMPH-induced state may influence motivation and drug-related assessments. The methodology described here may provide an easy and rapid way to investigate the determinants of AMPH-induced hypoactivity and acute withdrawal. PMID- 16364381 TI - Apple polygalacturonase inhibiting protein1 expressed in transgenic tobacco inhibits polygalacturonases from fungal pathogens of apple and the anthracnose pathogen of lupins. AB - Extracts from apple fruit (cultivar "Granny Smith") inhibited the cell-wall degrading polygalacturonase (PG) activity of Colletotrichum lupini, the causal agent of anthracnose on lupins, as well as Aspergillus niger PG. Southern blot analysis indicated that this cultivar of apple has a small gene family of polygalacturonase inhibiting proteins (pgips), and therefore heterologous expression in transgenic tobacco was used to identify the specific gene product responsible for the inhibitory activity. A previously isolated pgip gene, termed Mdpgip1, was introduced into tobacco (Nicotiana tabacum) by Agrobacterium mediated transformation. The mature MdPGIP1 protein was purified to apparent homogeneity from tobacco leaves by high salt extraction, clarification by DEAE Sepharose and cation exchange HPLC. Purified MdPGIP1 inhibited PGs from C. lupini and PGs from two economically important pathogens of apple trees, Botryosphaeria obtusa and Diaporthe ambigua. It did not inhibit the A. niger PG, which was in contrast to the apple fruit extract used in this study. We conclude that there are at least two active PGIPs expressed in apple, which differ in their charge properties and ability to inhibit A. niger PG. PMID- 16364382 TI - Conceptions and uses of public health ideas by New Zealand government policymakers: report on a five-agency survey. AB - OBJECTIVES: This article describes New Zealand government policymakers' awareness of, attitudes toward and self-reported use of ideas about the determinants of health. Some comparison with an earlier Canadian study is provided. METHODS: Employees with policymaking and advisory responsibilities in government departments of health, social development, housing, education and finance were surveyed. RESULTS: More than half of the respondents (58%) were familiar with ideas about health determinants. Eighty percent felt that health determinants should be considered in all government policy, but that more practical information on effective policy interventions is needed. Commitment to the idea that the economy should take precedence over reducing health inequalities was low. CONCLUSIONS: There is a demand for literature on health determinants that is sector specific and which provides practical and proven information about effective interventions that influence health. PMID- 16364383 TI - Risk of transfusion-transmitted hepatitis C virus in a tertiary hospital in Nigeria. AB - OBJECTIVES: To evaluate the risk of probable iatrogenic hepatitis C virus (HCV) infection following transfusion of donor blood that has not been screened for HCV. STUDY DESIGN: Prospective study. METHODS: Screening for human immunodeficiency virus and hepatitis B virus is routine in the University of Benin Teaching Hospital. HCV screening was performed on transfused bags of donor blood selected at random. The detection of anti-HCV was based on the principle of double antigen sandwich immunoassay, in which purified recombinant antigens are employed sufficiently to identify anti-HCV. The outcomes of interest included the proportion of HCV-positive units of transfused donor blood, the source of blood and the total number of units of blood processed in the hospital blood bank. RESULTS: A total of 4532 units of donor blood were procured in the blood bank. Of these, 4132 units were certified as fit for transfusion following the hospital protocols. The sources of the transfused blood samples were commercial blood donors [89.2% (n = 3687)] and targeted donation [10.8% (n = 445)]. One hundred and ninety-two transfused blood samples were randomly screened for HCV, and 3% (n = 6) were found to be positive (95% confidence intervals 0.007-0.06). The likely risk of iatrogenic transfusion-related HCV infection was estimated to be 129 cases/year at the present rate of utilization of donor blood at the University of Benin Teaching Hospital. CONCLUSION: There is a risk of iatrogenic transfusion transmitted HCV in the study hospital. Hospitals in Nigeria should screen for HCV prior to allogeneic transfusion, which may help in avoiding transfusion-related HCV and its probable long-term effects. PMID- 16364384 TI - Non-response in a survey of cardiovascular risk factors in the Dutch population: determinants and resulting biases. AB - BACKGROUND: The aim of the research was to study the determinants of participation in a health examination survey (HES) which was carried out in a population that previously participated in a health interview survey (HIS) of Statistics Netherlands, and to estimate the effect of non-participation on both the prevalence of the main HES outcomes (risk factors for cardiovascular disease) and on relationships between variables. METHODS: Logistic regression was used to study the determinants of participation in the HES (n=3699) by those who had previously participated in the HIS (n=12,786). Linear models were used to predict the main outcomes in non-participants of the HES. Item non-response was handled by multiple imputation. RESULTS: HES participants had a higher socio-economic status and comprised more 'worried well', while the rural population were less likely to participate in the HES. Most predicted values of outcomes in HES non participants differed from those in HES participants, but much of this was due to differences in the age and gender composition of both groups. Taking age and gender differences into account, most predicted values of outcomes in the entire HIS population were within the 95% confidence intervals of the HES values, with the exception of body height in men and high-density lipoprotein cholesterol, fasting glucose and body weight in women. These differences are most likely to be due to the higher socio-economic status of HES participants. Relationships between HIS variables did not change significantly when using HES participants alone compared with all HIS participants. CONCLUSIONS: Despite a high rate of non participation, some bias, mostly small, was seen in the prevalence rates of the main outcome variables. Bias in the relationships between variables was negligible. PMID- 16364385 TI - Rubella seroprevalence among first-grade primary school students in a district in Istanbul, Turkey. AB - OBJECTIVES: Rubella vaccination is not included in the national immunization programme of Turkey. However, it is on the market and is recommended, mainly by the private sector, at the age of 12-15 months. Introduction of the rubella vaccine without achieving high coverage has the risk of shifting the mean age of rubella infection among the unvaccinated population to reproductive ages. The aim of this study was to evaluate the seroprevalance of rubella and the factors associated with seropositivity among primary school students in Umraniye, a district of Istanbul. METHODS: Blood samples were collected from 489 students in 70 primary schools of Umraniye. Parents completed a concise questionnaire assessing sociodemographic characteristics and measles, mumps and rubella vaccination status. RESULTS: Among the students screened, 163 (33.3%, 95% confidence intervals 29.2-37.7%) were susceptible to rubella. When sociodemographic variables were evaluated, paternal education was the only variable associated with seronegativity. As the level of paternal education increased, seronegativity rates decreased. This finding may suggest that rubella vaccination is practiced more commonly among children whose fathers are better educated. CONCLUSIONS: Surveillance and control of rubella and congenital rubella syndrome must be set as public health priorities in Turkey, particularly given that rubella vaccination is practiced partially in the country. PMID- 16364386 TI - p38 Mitogen-activated protein kinase mediates IL-8 induction by the ribotoxin deoxynivalenol in human monocytes. AB - The effects of the ribotoxic trichothecene deoxynivalenol (DON) on mitogen activated protein kinase (MAPK)-mediated IL-8 expression were investigated in cloned human monocytes and peripheral blood mononuclear cells (PBMC). DON (250 to 1000 ng/ml) induced both IL-8 mRNA and IL-8 heteronuclear RNA (hnRNA), an indicator of IL-8 transcription, in the human U937 monocytic cell line in a concentration-dependent manner. Expression of IL-8 hnRNA, mRNA and protein correlated with p38 phosphorylation and was completely abrogated by the p38 MAPK inhibitor SB203580. DON at 500 ng/ml similarly induced p38-dependent IL-8 protein and mRNA expression in PBMC cultures from healthy volunteers. Significantly increased IL-6 and IL-1beta intracellular protein and mRNA expression was also observed in PBMC treated with DON (500 ng/ml) which were also partially p38 dependent. Flow cytometry of PBMC revealed that DON-induced p38 phosphorylation varied among individuals relative to both threshold toxin concentrations (25-100 ng/ml) and relative increases in percentages of phospho-p38(+) cells. DON-induced p38 activation occurred exclusively in the CD14(+) monocyte population. DON was devoid of agonist activity for human Toll-like receptors 2, 3, 4, 5, 7, 8 and 9. However, two other ribotoxins, emetine and anisomycin, induced p38 phosphorylation in PBMC similarly to DON. Taken together, these data suggest that (1) p38 activation was required for induction of IL-8 and proinflammatory gene expression in the monocyte and (2) DON induced p38 activation in human monocytes via the ribotoxic stress response. PMID- 16364387 TI - Proteins in the saliva of the Ixodida (ticks): pharmacological features and biological significance. AB - The saliva of ticks (Suborder Ixodida) is critical to their survival as parasites. A tick bite should result in strong responses from the host defence systems (haemostatic, immune and inflammatory) but tick saliva appears to have evolved to counter these responses. We review current knowledge of tick saliva components, with emphasis on those molecules confirmed to be present in the secreted saliva but including some that have only been confirmed to be present in salivary glands. About 50 tick saliva proteins that are well described in the literature are discussed. These saliva components include enzymes, enzyme inhibitors, amine-binding proteins and cytokine homologues that act as anti haemostatic, anti-inflammatory or immuno-modulatory agents. Sequence comparisons are illustrated. The importance of tick saliva and the significance of the findings to date are also discussed. PMID- 16364388 TI - Identification of a novel VP4 genotype carried by a serotype G5 porcine rotavirus strain. AB - Rotavirus genome segment 4, encoding the spike outer capsid VP4 protein, of a porcine rotavirus (PoRV) strain, 134/04-15, identified in Italy was sequenced, and the predicted amino acid (aa) sequence was compared to those of all known VP4 (P) genotypes. The aa sequence of the full-length VP4 protein of the PoRV strain 134/04-15 showed aa identity values ranging from 59.7% (bovine strain KK3, P8[11]) to 86.09% (porcine strain A46, P[13]) with those of the remaining 25 P genotypes. Moreover, aa sequence analysis of the corresponding VP8* trypsin cleavage fragment revealed that the PoRV strain 134/04-15 shared low identity, ranging from 37.52% (bovine strain 993/83, P[17]) to 73.6% (porcine strain MDR 13, P[13]), with those of the remaining 25 P genotypes. Phylogenetic relationships showed that the VP4 of the PoRV strain 134/04-15 shares a common evolutionary origin with porcine P[13] and lapine P[22] rotavirus strains. Additional sequence analyses of the VP7, VP6, and NSP4 genes of the PoRV strain 134/04-15 revealed the highest VP7 aa identity (95.9%) to G5 porcine strains, a porcine-like VP6 within VP6 genogroup I, and a Wa-like (genotype B) NSP4, respectively. Altogether, these results indicate that the PoRV strain 134/04-15 should be considered as prototype of a new VP4 genotype, P[26], and provide further evidence for the vast genetic and antigenic diversity of group A rotaviruses. PMID- 16364389 TI - Conserved cell cycle regulatory properties within the amino terminal domain of the Epstein-Barr virus nuclear antigen 3C. AB - The gammaherpesviruses Rhesus lymphocryptovirus (LCV) and Epstein-Barr virus (EBV) are closely related phylogenetically. Rhesus LCV efficiently immortalizes Rhesus B cells in vitro. However, despite a high degree of conservation between the Rhesus LCV and EBV genomes, Rhesus LCV fails to immortalize human B cells in vitro. This species restriction may, at least in part, be linked to the EBV nuclear antigens (EBNAs) and latent membrane proteins (LMPs), known to be essential for B cell transformation. We compared specific properties of EBNA3C, a well-characterized and essential EBV protein, with its Rhesus counterpart to determine whether EBNA3C phenotypes which contribute to cell cycle regulation are conserved in the Rhesus LCV. We show that both EBNA3C and Rhesus EBNA3C bind to a conserved region of mammalian cyclins, regulate pRb stability, and modulate SCF(Skp2)-dependent ubiquitination. These results suggest that Rhesus LCV restriction from human B cell immortalization is independent of the conserved cell cycle regulatory functions of the EBNA3C protein. PMID- 16364390 TI - The color appearance of stimuli detected via short-wavelength-sensitive cones for breast cancer survivors using tamoxifen. AB - This study compared color appearance data for two populations of middle-aged women: (1) women using the selective estrogen receptor modulator tamoxifen as adjuvant therapy following successful treatment for early-stage breast cancer, and (2) healthy amenorrheic (peri- or post-menopausal) control subjects not using any hormonally acting medications. A 3-alternative forced-response paradigm (with "blue", "lavender", or "white" as the allowable responses) was used for assessing the perceived color of 440-nm foveal test stimuli superimposed on a larger 3.6 log td 580-nm background. In contrast to the control subjects, the majority of tamoxifen users described the test stimuli as "white", whether at threshold or slightly suprathreshold. Threshold-sensitivity measurements themselves appeared less effective for distinguishing between subject groups. The results show that a simple color-naming procedure can reveal alterations of foveal short-wavelength sensitive cone-mediated visual function that are common among tamoxifen users. PMID- 16364391 TI - The accessibility of spatial channels for stereo and motion. AB - Using fractal noise images, we measured the dependence of D(min) on the spatial passband (spatial frequency and orientation) over which information was correlated either between the eyes for stereo or between sequential frames for motion. Without affecting the amplitude spectrum of the noise stimulus we used idealized filters to scramble the phase of components outside a pre-defined passband. Using a simple Gaussian model in which performance depends on the signal/noise within a restricted spatial region, we obtained estimates of the bandwidth of the narrowest underlying spatial frequency and orientation spectral region subserving these two comparable tasks. Spatial bandwidths varied with peak spatial frequency but were very broad approximating the spectrum of the stimulus itself. Orientation properties of the underlying mechanisms were isotropic. These results suggest that the independent activity of individual narrowband spatial channels is not perceptually accessible for these tasks. PMID- 16364392 TI - Disparity-defined objects moving in depth do not elicit three-dimensional shape constancy. AB - Observers generally fail to recover three-dimensional shape accurately from binocular disparity. Typically, depth is overestimated at near distances and underestimated at far distances [Johnston, E. B. (1991). Systematic distortions of shape from stereopsis. Vision Research, 31, 1351-1360]. A simple prediction from this is that disparity-defined objects should appear to expand in depth when moving towards the observer, and compress in depth when moving away. However, additional information is provided when an object moves from which 3D Euclidean shape can be recovered, be this through the addition of structure from motion information [Richards, W. (1985). Structure from stereo and motion. Journal of the Optical Society of America A, 2, 343-349], or the use of non-generic strategies [Todd, J. T., & Norman, J. F. (2003). The visual perception of 3-D shape from multiple cues: Are observers capable of perceiving metric structure? Perception and Psychophysics, 65, 31-47]. Here, we investigated shape constancy for objects moving in depth. We found that to be perceived as constant in shape, objects needed to contract in depth when moving toward the observer, and expand in depth when moving away, countering the effects of incorrect distance scaling (Johnston, 1991). This is a striking example of the failure of shape constancy, but one that is predicted if observers neither accurately estimate object distance in order to recover Euclidean shape, nor are able to base their responses on a simpler processing strategy. PMID- 16364393 TI - Quantifying facial expression recognition across viewing conditions. AB - Facial expressions are key to social interactions and to assessment of potential danger in various situations. Therefore, our brains must be able to recognize facial expressions when they are transformed in biologically plausible ways. We used synthetic happy, sad, angry and fearful faces to determine the amount of geometric change required to recognize these emotions during brief presentations. Five-alternative forced choice conditions involving central viewing, peripheral viewing and inversion were used to study recognition among the four emotions. Two alternative forced choice was used to study affect discrimination when spatial frequency information in the stimulus was modified. The results show an emotion and task-dependent pattern of detection. Facial expressions presented with low peak frequencies are much harder to discriminate from neutral than faces defined by either mid or high peak frequencies. Peripheral presentation of faces also makes recognition much more difficult, except for happy faces. Differences between fearful detection and recognition tasks are probably due to common confusions with sadness when recognizing fear from among other emotions. These findings further support the idea that these emotions are processed separately from each other. PMID- 16364394 TI - Holistic processing unites face parts across time. AB - When complementary halves of different familiar faces are combined into a new face, there is interference in the identification of either half. This "composite face effect" has been taken as strong evidence that faces are processed holistically. Here, we demonstrate that this effect can persist when the two parts of a face are separated by up to 80 ms of visual noise, showing that the parts of a face interact not only spatially but also temporally. We suggest that the processing underlying robust identification accepts an accumulation of evidence over time. PMID- 16364395 TI - Object discrimination in pigeons: effects of local and global cues. AB - We trained two pigeons to report whether a pair of differently colored 2-D objects had two target dots on either one or both of the objects. Follow-up tests disclosed that the colored regions surrounding the task-relevant targets were necessary, but not sufficient to support the birds' discrimination. Moreover, when local and global color cues provided contradictory information, pigeons failed to discriminate the stimuli, suggesting that the birds attended to both local and global information. Finally, one bird learned the object discrimination in the absence of differential color cues suggesting that, with suitable training, pigeon can attend to entire objects. PMID- 16364396 TI - Partial preservation of rod and cone ERG function following subretinal injection of ARPE-19 cells in RCS rats. AB - We quantified rod- and cone-related electroretinogram (ERG) responses following subretinal injections of the human-derived retinal pigment epithelial (hRPE) cell line ARPE-19 at age P23 to prevent progressive photoreceptor loss in the Royal College of Surgeons (RCS) rat. Culture medium-injected eyes served as sham controls. At P60, in comparison with sham-injected eyes, all recordings from hRPE injected eyes showed preserved scotopic a- and b-waves, oscillatory potentials, double-flash-derived rod b-waves and photopic cone b-waves, and flicker critical fusion frequencies and amplitudes. Although the actual preservation did not exceed 10% of a-wave and 20% of b-wave amplitude values in non-dystrophic RCS and deteriorated rapidly by P90, rod- and cone-related ERG parameters were still recordable up to P120 unlike the virtually unresponsive sham-injected eyes. PMID- 16364397 TI - The two-pulse experiment and cross-correlation. AB - Rashbass [Rashbass, C. (1970). The visibility of transient changes of luminance. Journal of Physiology, 210, 165-186] presented pairs of flashes having various contrasts separated by a delay, and found that the thresholds for detecting the pairs fell on an ellipse. He fit the data using a model that computed the filtered energy of the pulses. Although this Rashbass model is phase-insensitive, many other experimental results show that humans can perform phase-sensitive detection consistent with a template-matching mechanism. We show that an observer who uses a form of template-matching produces thresholds that fall on an ellipse, just like the Rashbass model. The results from two-pulse experiments are consistent with the idea that humans cross-correlate the stimulus (signal or noise) with a filtered version of the expected signal rather than the signal itself. In symbols, we propose that observers compute integral r(t)[s(t) *h(t)]dt where r(t) is the received stimulus on a given trial [s(t)+n(t) or n(t)], s(t) is the signal, h(t) is the visual filter, and * is convolution. PMID- 16364398 TI - The impact of selected water quality parameters on the inactivation of Bacillus subtilis spores by monochloramine and ozone. AB - Selected water quality parameters-pH, dissolved organic carbon, turbidity (NTU), and temperature-were tested for their potential effects on ozone and monochloramine inactivation of Bacillus subtilis spores. In oxidant demand-free phosphate-buffer, temperature had the strongest influence on inactivation kinetics when using ozone, pH had a smaller but significant impact on B. subtilis spore inactivation with both monochloramine and ozone. Where monochloramine was applied, modeling and experimental measurements confirmed that dichloramine levels were too low to produce significant inactivation effects under these experimental conditions. It was demonstrated that oxidant demand-free phosphate buffer may not be an adequate environmental analogue for inactivation responses in natural waters. PMID- 16364399 TI - Organochlorinated pesticide multiresidues in surface sediments from Beijing Guanting reservoir. AB - For the analysis of organochlorine pesticides (OCPs) in sediments and water, a multiresidue analysis method based on gas chromatography with electron capture detection (GC/ECD) was developed. Solid-phase extraction (SPE) using Oasis HLB cartridges was also applied in sample extraction. Mean recoveries were satisfactory with 72-103% and 71-103% for water and sediment, respectively. Twenty-one OCPs were analyzed in water and surface sediment samples from Beijing Guanting reservoir. Total concentrations of 21 OCPs were 16.7-791 ng/l (mean 234 ng/l), 275-1,600 ng/l (mean 644 ng/l), 5250-33,400 ng/kg (mean 13,000 ng/kg) in surface water, pore water and sediment (dry weight), respectively. P, p'-DDE, delta-HCH, aldrin, gamma-HCH and beta-HCH are the most abundant compounds in water while p, p'-DDE, o, p'-DDT, beta-HCH, delta-HCH, p, p'-DDT and aldrin accounted for about 85% of total 21 OCPs in sediment in Beijing Guanting reservoir. Concentrations of OCPs were high both in water and sediment samples in Beijing Guanting reservoir for municipal water supply. The data provides information on the levels and sources of OCPs in Guanting reservoir. The study indicated that measures should be taken to decrease OCPs residues in order to improve the quality of municipal water supply in Beijing. PMID- 16364400 TI - A Bayesian method to estimate proportional payments of users in a wastewater treatment plant. AB - A simple Bayesian method to estimate proportional payments for using or constructing a wastewater treatment plant is presented. The method addresses the usually unknown individual participation in global pollution, of municipal and industrial wastewaters that are normally discharged into the same collecting pond. The individual contributions are calculated from the discharged volume of each source and its pollution percentage in the wastewater volume previous to the disposal. According to the method presented here, the costs in building and/or running the plant can be covered by proportional individual payments based on the individual contributions, in a more even way. The simplicity of the model allows to assess different strategies for the participants in a wastewater treatment system. A real case study in the Izta-Popo National Park, located in the Mexican States of Puebla, Mexico and Morelos, is analyzed. PMID- 16364401 TI - Influence of internal biogas production on hydrodynamic behavior of anaerobic fluidized-bed reactors. AB - Predictive models for describing the hydrodynamic behavior (bed-expansion and bed pressure gradient) of a three-phase anaerobic fluidized bed reactor (AFBR) was developed according to wake theory together with more realistic dynamic bed expansion experiments (with and without internal biogas production). A reliable correlation equation for the parameter k (mean volume ratio of wakes to bubbles) was also established, which is of help in estimating liquid hold up of fluidized beds. The experimental expansion ratio of three-phase fluidized beds (E(GLS)) was approximately 18% higher than that of two-phase fluidized beds (E(LS)); whereas the experimental bed-pressure gradient of the former [(-DeltaP/H)(GLS)] was approximately 9.3% lower than that of the latter [(-DeltaP/H)(LS)]. Both the experimental and modeling results indicated that a higher superficial gas velocity (u(g)) gave a higher E(GLS) and a higher E(GLS) to E(LS) ratio as well as a lower (-DeltaP/H)(GLS) and a lower (-DeltaP/H)(GLS) to (-DeltaP/H)(LS) ratio. As for the operation stability of the AFBR, the sensitivity of u(g) to expansion height (H(GLS)) and (-DeltaP/H)(GLS) is between the sensitivity of superficial liquid velocity and biofilm thickness. The model predictions of E(GLS), (-DeltaP)(GLS), and (-DeltaP/H)(GLS) agreed well the experimental measurements. Accordingly, the predictive models accounting for internal biogas production described fairly well the hydrodynamic behavior of the AFBR. PMID- 16364402 TI - Degradation of Pb--EDTA complex by a H(2)O(2)/UV process. AB - The degradation of PbEDTA in aqueous solution by a H(2)O(2)/UV process was studied. The effect of H(2)O(2) content, pH of the solution and the presence of nitrate were investigated. PbEDTA degradation by a H(2)O(2)/UV process was shown to be accompanied by simultaneous lead precipitation. PbEDTA was decomposed rapidly in acidic solutions while lead precipitation was achieved only when the pH of the solution was higher than 6. The presence of nitrate in significant amounts (0.04 M) inhibited remarkably the degradation of the complex and metal precipitation. The degradation of CdEDTA and ZnEDTA was also studied. It was found that the decomposition of metal-EDTA complex and metal removal by the H(2)O(2)/UV process depend greatly on the nature of the metal. CdEDTA and ZnEDTA were decomposed rapidly but metal precipitation was not achieved. The major by products of the degradation of metal-EDTA complexes observed were nitrilotriacetic acid (NTA), iminodiacetic acid (IDA), oxalic acid and nitrate. PMID- 16364403 TI - Uptake and transformation of phenol and chlorophenols by hairy root cultures of Daucus carota, Ipomoea batatas and Solanum aviculare. AB - Hairy root cultures of Daucus carota L., Ipomoea batatas L. and Solanum aviculare Forst were investigated for their susceptibility to the highly toxic pollutants phenol and chlorophenols and for the involvement of inherent peroxidases in the removal of phenols from liquid media. Roots of D. carota grew normally in medium containing 1000 micromol l(-1) of phenol, whilst normal growth of roots of I. batatas and S. aviculare was only possible at levels up to 500 micromol l(-1). In the presence of chlorophenols, normal root growth was possible only in concentrations not exceeding 50 micromol l(-1), except for I. batatas which was severely affected at all concentrations. Despite the reduction in biomass, the growth of S. aviculare cultures was sustained in medium containing up to 2000 micromol l(-1) of phenol or 2-chlorophenol, and up to 500 micromol l(-1) of 2,6 dichlorophenol. The amounts of phenol removed by the roots within 72 h of treatment were 72.7%, 90.7% and 98.6% of the initial concentration for D. carota, I. batatas and S. aviculare, respectively. For the removal of 2,6-dichlorophenol the values were, respectively, 83.0%, 57.7% and 73.1%. Phenols labelled with 14C were absorbed by the root tissues and condensed with highly polar cellular substances as well as being incorporated into the cell walls or membranes. The results suggest that S. aviculare, an ornamental plant, would be best suited for remediation trials under field conditions. PMID- 16364404 TI - Isomeric analysis of BTEXs in the atmosphere using beta-cyclodextrin capillary chromatography coupled with thermal desorption and mass spectrometry. AB - An analytical method capable of determining trace levels of BTEX-aromatics (benzene, toluene, ethylbenzene, m-, p- and o-xylenes) in the atmosphere with as high resolution as possible has been developed. The method is based on the preconcentration of air samples using a multibed tube (Carbopack C, Carbograph 1) at ambient temperature, followed by thermal desorption, and analysis of aromatic species by a beta-cyclodextrin capillary chromatography coupled with mass spectrometry. The resolution achieved was sufficient for individual separation of BTEXs as well as m- and p-xylenes. The BTEX-ratios have been determined in an air tunnel and in on-road, suburban and rural forest atmosphere. The ethylbenzene/m xylene ratios could provide a deep insight into anthropogenic related NMHC patterns at different locations and under different meteorological conditions and may reflect photochemical processes in the best way. PMID- 16364405 TI - Adaptive origins of primates revisited. AB - Interpretation of the adaptive profile of ancestral primates is controversial and has been constrained for decades by general acceptance of the premise that the first primates were very small. Here we show that neither the fossil record nor modern species provide evidence that the last common ancestor of living primates was small. Instead, comparative weight distributions of arboreal mammals and a phylogenetic reconstruction of ancestral primate body mass indicate that the reduction of functional claws to nails -- a primate characteristic that had up until now eluded satisfactory explanation - resulted from an increase in body mass to around 1000 g or more in the primate stem lineage. The associated shift to a largely vegetarian diet coincided with increased angiosperm diversity and the evolution of larger fruit size during the Late Cretaceous. PMID- 16364406 TI - ESR and U-series analyses of enamel and dentine fragments of the Banyoles mandible. AB - The Banyoles mandible presents a puzzle. Its anatomy has been described as pre Neandertal, but the travertine in which it was found has been dated to 45,000 +/- 4000 years. By this time, any pre-Neandertals had supposedly been absent from the European fossil record for more than 100,000 years. It was therefore proposed that the age of the travertine may represent a minimum age estimate, with the mandible possibly having been reworked from older deposits. We carried out a non destructive ESR analysis of an enamel fragment removed from a molar and performed a series of in situ laser ablation U-series analyses on dentine fragments adjacent to the enamel piece. The analyses resulted in an apparent combined ESR-U series age of 66,000 +/- 7000 years. The encasing travertine matrix was also analyzed for U-series isotopes and showed signs of U-mobilization. It cannot be excluded that the travertine matrix is older than the previously determined age. If the mandible was not reworked, then the combined ESR-U-series result on the tooth enamel would give its best age estimate. If, on the other hand, the mandible was reworked from another deposit, the actual ESR-U-series age will depend on the external dose rate from the previous matrix and the depth of its burial, which controls the degree of the attenuation of the cosmic dose rate over time. Considering a range of possible burial histories, the mean age of the mandible would lie somewhere between the combined ESR-U-series age and the previously determined age of the travertine matrix. Regarding the morphology of the mandible, a review of its features in the context of larger Neandertal samples indicates that the anatomy of the specimen is not incompatible with such a young age determination, although it further highlights morphological variation in the late Neandertal sample. PMID- 16364407 TI - Simulating effects of fire disturbance and climate change on boreal forest productivity and evapotranspiration. AB - We used a terrestrial ecosystem process model, BIOME-BGC, to investigate historical climate change and fire disturbance effects on regional carbon and water budgets within a 357,500 km(2) portion of the Canadian boreal forest. Historical patterns of increasing atmospheric CO2, climate change, and regional fire activity were used as model drivers to evaluate the relative effects of these impacts to spatial patterns and temporal trends in forest net primary production (NPP) and evapotranspiration (ET). Historical trends of increasing atmospheric CO2 resulted in overall 13% and 5% increases in annual NPP and ET from 1994 to 1996, respectively. NPP was found to be relatively sensitive to changes in air temperature (T(a)), while ET was more sensitive to precipitation (P) change within the ranges of observed climate variability (e.g., +/-2 degrees C for T(a) and +/-20% for P). In addition, the potential effect of climate change related warming on NPP is exacerbated or offset depending on whether these changes are accompanied by respective decreases or increases in precipitation. Historical fire activity generally resulted in reductions of both NPP and ET, which consumed an average of approximately 6% of annual NPP from 1959 to 1996. Areas currently occupied by dry conifer forests were found to be subject to more frequent fire activity, which consumed approximately 8% of annual NPP. The results of this study show that the North American boreal ecosystem is sensitive to historical patterns of increasing atmospheric CO2, climate change and regional fire activity. The relative impacts of these disturbances on NPP and ET interact in complex ways and are spatially variable depending on regional land cover and climate gradients. PMID- 16364408 TI - Assessment of heavy metal accumulation in two species of Tillandsia in relation to atmospheric emission sources in Argentina. AB - The ability of Tillandsia capillaris Ruiz and Pav. f. capillaris and Tillandsia permutata A. Cast. to accumulate heavy metals was evaluated in relation to potential atmospheric emission sources in Argentina. The sampling areas (n=38) were chosen in the province of Cordoba, located in the center of Argentina, and categorized according to land use, anthropogenic activities and/or distance to potential heavy metal emission sources. In each sampling site, pools of 40-50 individuals of each species were made from plants collected along the four cardinal directions. The concentrations of V, Mn, Fe, Co, Ni, Cu, Zn, Pb and Br of these samples were measured by Total Reflection X-Ray Fluorescence (TXRF) analysis with Synchrotron Radiation. Each species was submitted to a cluster analysis in order to discriminate different groups of heavy metals as tracers of natural or anthropogenic sources. A Contamination Factor (CF) was calculated using the concentrations of the elements in each sample compared to their concentrations in the control samples. Finally, the rank coefficients of correlation between the CFs and the categorical variables characteristic of each site (land use and anthropogenic load) were analyzed. A positive correlation was found for T. capillaris between the CFs of V, Mn, Co, Ni, Cu and Zn and the urban industrial category, whereas the CF values for Zn and Pb were positively correlated with the road category. In T. permutata there was a positive correlation between the CF of Zn and the urban-industrial category and the CF of Pb with the road category. We therefore conclude that T. capillaris is a more efficient metal accumulator in passive biomonitoring studies. PMID- 16364409 TI - Elemental gaseous mercury flux at the water/air interface over the Negro River basin, Amazon, Brazil. AB - The amount of dissolved gaseous mercury (DGM) released annually into the atmosphere by water bodies in each of the 18 major sub-basins of the Negro River tributaries has been estimated for the wet season and adopted for the entire year. Using Remote Sensors (RS) and the Geographic Information System (GIS), an estimate was made for the total flooded area during the period of the wet season (May-July), which corresponds to around 10% of the total basin area. The estimates of DGM evasive flux values were made during four scientific campaigns and ranged from 0.09 to 14 mug m(-2) y(-1). Mercury wet deposition to the same area was estimated using average values of the metal concentration in the rainwater (9.8 ng L(-1)), which varies from 17 to 27 microg m(-2) year(-1). Considering the flooded area of the Negro River Basin as 69,000 km(2), the total amount of mercury emitted by all bodies of water reaches nearly 0.26 ton year( 1), which represents not more than 2% of the total mercury found in wet deposition in this same area. PMID- 16364410 TI - Eco-efficiency evaluation of a smart window prototype. AB - An eco-efficiency analysis was conducted using indicators suitably defined to evaluate the performance of an electrochromic window acting as an energy saving component in buildings. Combining the indicators for various parameters (control scenario, expected lifetime, climatic type, purchase cost) significant conclusions are drawn for the development and the potential applications of the device compared to other commercial fenestration products. The reduction of the purchase cost (to 200 euros/m2) and the increase of the lifetime (above 15 years) are the two main targets for achieving both cost and environmental efficiency. An electrochromic device, implemented in cooling dominated areas and operated with an optimum control strategy for the maximum expected lifetime (25 years), can reduce the building energy requirements by 52%. Furthermore, the total energy savings provided will be 33 times more than the energy required for its production while the emission of 615 kg CO2 equivalent per electrochromic glazing unit can be avoided. PMID- 16364411 TI - Gender of offspring and maternal ovarian cancer risk. AB - OBJECTIVE: A single live birth compared to nulliparity significantly reduces the risk for ovarian cancer, but exactly how pregnancy reduces ovarian cancer risk is unknown. We sought to determine whether offspring gender, which differentially alters maternal hormonal milieu, may be associated with maternal ovarian cancer risk. METHODS: Parous women (n = 511) with incident ovarian cancer were compared to parous community controls (n = 1136) participating in a population-based case control study of ovarian cancer (Delaware Valley, 1994-1998). In subgroup specific models for women with one, two, or three births, multivariate logistic regression was used to assess the relationship between ovarian cancer and offspring gender, adjusting for age, race, education, oral contraceptives, breast feeding, tubal ligation, and ovarian cancer family history. RESULTS: Compared to having all girls, women with all boys tended to have a reduced risk of ovarian cancer (OR = 0.80 95% CI: 0.58, 1.10), while women with boys and girls conferred the greatest protection (OR = 0.58, 95% CI: 0.43, 0.79). Among women with two births, the association was observed for those with one boy and one girl (OR = 0.63, 95% CI: 0.40, 1.00), but not for those with two male offspring (OR = 1.12, 95% CI: 0.68, 1.85). This result was consistent among women with three births (OR = 0.42, 95% CI: 0.21, 0.84; OR = 0.47, 95% CI: 0.23, 0.95; OR = 0.49, 95% CI: 0.20, 1.21; for one, two, and three boys, respectively, compared to all girls). CONCLUSION: Compared to having all girls, bearing both male and female offspring may be associated with a decrease in maternal ovarian cancer risk, although the biologic relevance of this observation is unclear. PMID- 16364412 TI - Saphenous vein sparing during inguinal lymphadenectomy to reduce morbidity in patients with vulvar carcinoma. AB - OBJECTIVES: To compare short- and long-term morbidity associated with saphenous vein sparing versus ligation during inguinal lymphadenectomy for vulvar carcinoma. METHODS: A retrospective evaluation of patients with carcinoma of the vulva that underwent inguinal lymphadenectomy was performed. Operative reports were evaluated and patients were divided into those who had sparing of the saphenous vein versus ligation. Postoperative short- and long-term complications were compared between the two groups using Pearson chi squared analysis. RESULTS: There were a total of 49 inguinal lymphadenectomies performed on 29 patients. The saphenous vein was spared in 18 (37%) groin dissections compared to 31(63%) in which the saphenous vein was ligated. The two groups were similar in regards to clinical characteristics. All patients received closed suction drains and prophylactic antibiotics. Median number of nodes dissected was similar. Cellulitis was more common in the vein-ligated group compared to the vein-spared group (45% vs. 0%; P < 0.001). Wound breakdown occurred in 25% of dissections where the saphenous vein was ligated versus 0% in dissections where the vein was spared (P = or < 0.02). Short-term edema (< or = 6 months) was similar between vein-ligated and vein-spared groups (67% vs. 72%, P < 1.0). Subsequently, chronic lymphedema (> 6 months) persisted in 38% of the vein-ligated group compared to 11% in the vein-spared group (P < 0.05). The incidence of recurrent disease was similar in both groups (19.3 % vs. 22.2% P < 0.1). CONCLUSIONS: Routine preservation of the saphenous vein during inguinal lymphadenectomy for vulvar carcinoma may reduce the incidence of wound cellulitis, wound breakdown, and chronic lymphedema. PMID- 16364413 TI - Platinum(IV) complex with adamantylamine overcomes intrinsic resistance to cisplatin in ovarian cancer cells. AB - OBJECTIVES: The resistance of tumor cells to cisplatin remains a major cause of treatment failure in cancer patients. In this study, the ability of Pt(IV) complex with adamantylamine-LA-12 and its reduced counterpart with lower oxidation state Pt(II)-LA-9 to overcome intrinsic cisplatin resistance was investigated. METHODS: The ovarian adenocarcinoma SK-OV-3 cells were exposed to cisplatin, LA-9, or LA-12 for 72 h and the effects of drug concentrations that caused 10% or 50% inhibition of cell proliferation were determined. After 24-72 h of sustained exposure viability, apoptosis and inhibition of proliferation were analyzed. DNA synthesis and cell cycle analysis were performed simultaneously in order to determine the modulation of cell cycle after platinum complexes treatment. RESULTS: Lung Resistance-related Protein (LRP/MVP) was detected in SK OV-3 cells but not in the other two ovarian cancer lines with different sensitivity to cisplatin. LRP/MVP overexpression may be an important factor contributing to intrinsic cisplatin resistance. Interestingly, Pt(IV) complex-LA 12 had approximately 2.7-fold lower IC(50) concentration than LA-9 or cisplatin in SK-OV-3 cells. Moreover, LA-12 caused persistent accumulation of cells in S phase of the cell cycle while LA-9 and cisplatin treatment-induced S-phase arrest was transient and shifted to G(2)/M-phase at later intervals. Apoptosis seemed to be not the dominant type of cell death caused by such the derivatives, but it was the most intensive after LA-12 treatment. CONCLUSIONS: We found strong differences between effects of Pt(IV) complex-LA-12 and Pt(II) derivatives-LA-9 and cisplatin on cytokinetic parameters. Overall, LA-12 but not its reduced Pt(II) counterpart LA-9 is the compound effective in p53 null human ovarian cancer cells and it is able to overcome intrinsic cisplatin resistance in these cells. PMID- 16364414 TI - Effect of metabotropic glutamate receptor agonists and signal transduction modulators on feeding by a caterpillar. AB - Feeding in codling moth caterpillars was induced by the general glutamate receptor activator monosodium glutamate (MSG) and by three different mGluR agonists known to specifically stimulate different classes of vertebrate metabotropic glutamate receptors, including: (1S,3R)-ACPD, which stimulates group I mGluRs (2R,4R)-APDC, which stimulates group II mGluRs and L-AP4, which stimulates some group III mGluRs. Experiments exposing larvae to combinations of specific mGluR agonists and specific signal transduction modulators suggest that each tested mGluR uses a different signaling pathway. First, feeding stimulatory effects of (1S,3R)-ACPD were abolished by phospholipase C inhibitor, U 73122, but remained unaffected by adenylate cyclase activator, NKH 477, or phosphodiesterase inhibitor, Rolipram. Second, (2R,4R)-APDC induced feeding in presence of U 73122 or Rolipram, but lost its feeding stimulatory effects in presence of NKH 477. Finally, L-AP4 did not induce feeding in presence of Rolipram, but maintained its feeding stimulatory effects in presence of U 73122 or NKH 477. The activity of the general glutamate receptor activator MSG was abolished by NKH 477, and Rolipram. U 73122 did not affect MSG-stimulated feeding. These results suggest that transduction of MSG taste in the codling moth caterpillar relies mostly on cAMP-dependent signaling pathways. PMID- 16364415 TI - Changes in French people's misconceptions about hepatitis C, 1997-2003. AB - OBJECTIVE: The French health authorities instituted in 1999 a campaign to educate physicians, high-risk populations, and the public about hepatitis C (HC). To what extent has knowledge about HC increased among members of the French public? METHOD: A convenience sample of 706 French adults, aged 18 to 87, completed in 2003 a questionnaire designed to assess their knowledge of HC. Their responses were compared to those in 1997 of a different group of French adults on an identical questionnaire. RESULTS: Between 1997 and 2003, knowledge significantly improved on 13 of the 26 items for which strong expert-public differences were found in 1997 and significantly worsened on 3 items. CONCLUSIONS: France's public education campaign led to only a limited increase in knowledge about HC. Further public education is needed. PMID- 16364416 TI - Breeding soundness and libido examination of Belgian Blue and Holstein Friesian artificial insemination bulls in Belgium and The Netherlands. AB - Data on breeding soundness and libido evaluations in Belgian Blue (BB) bulls are scarce. The present study compared results of breeding soundness and libido evaluations of young BB bulls to young Holstein Friesian (HF) bulls prior to acceptance into an AI program. Breed differences for breeding soundness exist between BB and HF bulls, as 93.7% of the young BB bulls failed the breeding soundness examination (BSE) compared to 59.3% of the HF bulls (P=0.0005). Within the BB breed, differences were present between bulls of different ages, and bull selection for better fertility with increasing age apparently influenced the results. The number of reasons for which bulls failed the test differed between the age groups in the BB breed, whereas a tendency for more failure reasons in the BB breed was noticed in the breed comparison. The most important reasons for failure were sperm morphology and scrotal circumference (SC), but far more BB bulls failed for these traits compared to the HF breed (82.8% versus 56.0% and 43.8% versus 17.6% in the BB and the HF breed for sperm morphology (P=0.0005) and SC (P<0.0001), respectively). The high proportion of BB bulls with a substandard SC and poor sperm morphology might suggest an increased prevalence of testicular hypoplasia or degeneration within this breed. Concerning libido, the reaction time did not differ either between breeds or between age groups within the BB breed, whereas mounting enthusiasm, although not different between the two breeds, did decline with increasing age, probably due to the greater mating experience of the older bulls. All in all, libido did not seem to be different between the breeds. PMID- 16364417 TI - Validation of a rapid, large-scale assay to quantify ATP concentration in spermatozoa. AB - Quantification of ATP content in spermatozoa is a useful assay for evaluating sperm function; however, most detection methodology relies on assessing single samples. We have developed and validated a highly repeatable assay that permits simultaneous measurement of up to 78 samples. A key feature of this assay includes combination of a phosphatase inhibition and ATP extraction step that permits maximal detection of ATP and sample storage at -20 degrees C prior to assay. The assay was validated for spermatozoa from three different species, including turkey, rooster and boar. The sensitivity of the assay differed between avian and mammalian spermatozoa, with 2.5 x 10(6) spermatozoa being the lowest number of turkey and rooster spermatozoa that could be assayed compared to 2.5 x 10(5) boar spermatozoa. Concentrations of ATP in fresh turkey semen ranged from 2.14 to 15.6 nmol/10(9) spermatozoa; similarly, freshly collected rooster semen contained from 2.16 to 21.4 nmol ATP/10(9) spermatozoa. Evaluation of turkey semen that had been stored at 4 degrees C for 24 h revealed a decline in ATP concentrations (2.35 +/- 0.34 nmol ATP/10(9) spermatozoa). Likewise, cryopreserved rooster spermatozoa contained lower concentrations of ATP (0.05 +/- 0.01 nmol ATP/10(9) spermatozoa) than non-stored spermatozoa. Boar spermatozoa contained similar concentrations of ATP, whether fresh (74.2 +/- 8.1 pmol ATP/10(6) spermatozoa), stored for 1 day (77.0 +/- 8.1 pmol ATP/10(6) spermatozoa) or 5 days (81.96 +/- 8.1 pmol ATP/10(6) spermatozoa). For all three species, assay variation was low (inter-assay, 0.66-1.9% CV; intra-assay, 1.3% CV). PMID- 16364418 TI - The effect of a monensin controlled-release capsule on the incidence of retained fetal membranes, milk yield and reproductive responses in Holstein cows. AB - The objective of this study was to determine the effect of a monensin controlled release capsule on the incidence of retained fetal membranes (RFM), milk yield and conception rate at first service (CRFS). The study was conducted during May to August 2004 in a large dairy herd in Chihuahua, Mexico. Cows (n = 2,025) from 20 to 30 d before expected parturition were randomly assigned to either a treatment or a control group. The treated group (n = 1,010) received orally a capsule of monensin that released 335 mg/d for 95 d. Control cows (no capsule, n = 1,015) were randomly assigned and matched by parity. Outcome variables were the incidence of RFM, first three test day milk and CRFS. Monensin decreased the incidence of RFM by 3.9% only in multiparous cows. Therefore, multiparous cows treated with monensin were 0.78 times (95% C.I. = 0.71-0.86) as likely as control multiparous cows to develop RFM. Overall, milk yield was negatively affected by the RFM condition, but it was improved by monensin only in multiparous cows with RFM at test day 1. Conception rate at first service was depressed by 10.6% in multiparous cows with RFM. However, in RFM cows treated with monensin, CRFS improved to the level of treated cows without RFM. PMID- 16364419 TI - Curing-light attenuation in filled-resin restorative materials. AB - OBJECTIVE: To characterize the attenuation of the curing light in filled resin restorative materials (FRRMs) to aid understanding of curing depth. MATERIALS AND METHODS: One hundred and eighty materials of various shades from several manufacturers were tested in various ways. One set (66 materials) was used to determine the applicability of Lambert's Law using a quartz-tungsten-halogen curing light (Optilux 400, Demetron Research) by measuring the transmitted light with a dental radiometer (Cure Rite, EFOS) for successive thicknesses of ground 10mm diameter specimens from 3 to 0.5 mm in 0.5 mm steps. A second set (17 materials) were similarly tested with separate specimens from 1 to 5mm in thickness using a transmission densitometer (DT1405, RY Parry) fitted with a curing-light dichroic filter. For a third (overlapping) set (165 materials), the 1 mm pure (reflectance-free) optical density (D1 value) was determined from two specimens, approximately 1 and approximately 2 mm thick using the densitometer as above. From D1 the critical thickness (x(CRIT)), identified as depth of cure (DoC) for an excess surface exposure factor of 2, was calculated. RESULTS: Lambert's Law was found to hold with no evidence of appreciable differential absorption effects. Attenuation coefficient and D1 were significantly correlated (P < 1 x 10(-13)). D1 varied between about 0.23 and 0.72, for corresponding x(CRIT) values of 1.3 and 0.4 mm. There was no correlation between D1 and reflectance (P > 0.09), and no systematic effect due to shade letter, but a highly significant (P < 7.5 x 10(-8)), but weak (-0.066 mm/unit), correlation between shade number and D1. SIGNIFICANCE: Depth of cure can be calculated directly from the D1 value determined via simple optical density measurements on two specimens providing that (a) an irradiation time can be determined for the surface of a specimen to be "sufficiently" cured (i.e. for DoC = 0 precisely), and (b) an excess internal surface exposure ratio can be chosen such that the corresponding DoC is attained in a practicable irradiation time. PMID- 16364420 TI - Extracellular environment as one mediator of blue light-induced mitochondrial suppression. AB - OBJECTIVES: The current study tested the hypothesis that the extracellular environment mediates mitochondrial suppression of oral epithelial cells and fibroblasts by blue light. METHODS: We exposed Balb fibroblasts (Balb), normal human epidermal keratinocytes (NHEK), and oral squamous carcinoma cells (OSC2) to blue light (30-120J/cm2) in different cell-culture media and in phosphate buffered saline (PBS). Mitochondrial activity (MTT method) was used to assess cellular response 72 h post-light exposure. Cell-culture media were replaced or supplemented before or after light exposure to assess the variables of exposure time and medium degradation as mediators of blue light-induced effects. RESULTS: Mitochondrial activity of NHEK was not suppressed by exposure to blue light regardless of extracellular conditions. The mitochondrial activity of OSC2 and Balb cells was suppressed most when cells were exposed to light in cell-culture medium (versus PBS). Blue light suppressed mitochondrial activity more when irradiated medium remained in contact with the cells at least 1h, indicating a time-dependence of the medium effects. Neither a replacement nor a supplementation of medium components reduced blue light-induced mitochondrial suppression. SIGNIFICANCE: Our results suggest that tissue environments influence cellular responses to blue light and that these environments should be considered when assessing any biological effects of blue light during the photopolymerization of restorative resins. PMID- 16364421 TI - Fatigue testing of enamel bonds with self-etch and total-etch adhesive systems. AB - OBJECTIVE: Etching of enamel by self-etching adhesive systems is not as pronounced as with phosphoric acid employed with most total-etch adhesive systems. This may result in differences in the effectiveness of the bonds for the two types of systems. The aim of this study was to compare two such systems by bond strength and fatigue testing. MATERIALS AND METHODS: Shear bond strengths for Single Bond (SB) and Adper Prompt-L-Pop (PLP) were obtained on bovine enamel surfaces. Fatigue measurements were made with the same test fixtures using cyclical loading at 2 Hz for a maximum of 10(5) cycles or until the bond failed. Four selected loads between 40 and 70% of the shear bond strength were tested for each adhesive and the number of cycles to failure was recorded. S-N curves were constructed from the data and fatigue stress limits were determined. The Mann Whitney U-test and t-tests were used for statistical comparisons of the results. SEM analysis of resin tag formation into enamel surfaces was carried out. RESULTS: A significantly greater bond strength was found for SB (25.3 MPa) than for PLP (19.2). Sustainable stresses after 10(5) load cycles were substantially lower than the corresponding shear bond strengths and fatigue limits were found to be 8.4 MPa for PLP and 14.6 MPa for SB. The ratio of fatigue limit to bond strength was less for PLP (44%) than for SB (57%). Resin tag penetration into enamel was substantially less for PLP than for SB. SIGNIFICANCE: Fatigue testing in conjunction with bond strength testing can provide a better means for assessing the performance of adhesive systems used for bonding to enamel. PMID- 16364422 TI - Effect of temperature on the silane coupling agents when bonding core resin to quartz fiber posts. AB - OBJECTIVES: To evaluate the effect of different silane agents and air-drying temperatures on bond strength of translucent quartz fibre posts to composite resin. METHODS: The post surface was etched with 10 vol% hydrogen peroxide for 20 min. A two-liquid coupling agent containing 4-methacryolxyethyl trimellitate anhydride (4-META) and gamma-trimethoxysilyil propyl methacrylate (gamma-MPTS) and two pre-hydrolyzed single component silanes containing 3 methacryloxypropyltrimethoxysilane (3-MPS) and glycid-oxi-propyl-trimetil-oxi silane (GPS), respectively, were used for treating the fiber posts. Two different post-silanization drying temperatures were applied (21 and 38 degrees C). A dual cure composite resin (Core Paste XP) was selected to build-up the core around posts, obtaining cylindrical specimens that were serially cut in beams and subsequently loaded in tension (muTBS) at a cross-head speed of 1 mm/min until failure. Bond strength data were statistically analyzed by two-way ANOVA and Student-Newman-Keuls tests (alpha=0.05). RESULTS: Warm air-drying determined significantly higher bond strengths (p<0.001) for glycid-oxi-propyl-trimetil-oxi silane (11.6 MPa) and 4-methacryolxyethyl trimellitate anhydride/gamma trimethoxysilyil propyl methacrylate silane (11.7 MPa). These two systems exhibited lower bond strengths (6.9 and 8.8 MPa, respectively) than 3 methacryloxypropyltrimethoxysilane (11.0 MPa) when dried at 21 degrees C. No statistical differences were recorded for 3-methacryloxypropyltrimethoxysilane when drying at 21 or 38 degrees C. SIGNIFICANCE: The composition of the silane coupling agent in terms of acidic content, solvent rate or degree of hydrolysis may influence resin/post bond strength when dried at 21 degrees C. Drying at 38 degrees C most likely facilitates the evaporation of solvents present in the silane agent, resulting in increased bond strength of the composite resin to the fiber post. PMID- 16364423 TI - An investigation into the use of an anaerobic adhesive with two commercially available orthodontic brackets. AB - OBJECTIVES: The two objectives of this experiment were to determine the surface temperature of enamel following acid etching, rinsing and drying, and to see whether two commercially available orthodontic brackets could be bonded to enamel using an anaerobic adhesive. METHODS: Enamel surface temperature was determined in vivo using a surface temperature probe on a total of 60 patients. Stainless steel orthodontic brackets were bonded to human enamel using an anaerobic adhesive and a control orthodontic adhesive. The enamel was etched prior to bonding either with a solution of 37% o-phosphoric acid or, in the case of the anaerobic adhesive specimens, with a solution of 37% o-phosphoric acid containing copper (II) chloride. After bench curing the specimens were shear bond tested to failure and the load at debond recorded in each case. RESULTS: The bond test results were analyzed using median force to debond (N) and 95% confidence intervals, Kaplan-Meier survival probabilities and log-rank tests. CONCLUSIONS: After etching rinsing and drying the enamel surface temperature ranged from 21.54 to 24.19 degrees C, which is within the range suitable for anaerobic adhesive use. Bond testing to failure demonstrated that bracket base design affected the measured force to debond with both the anaerobic adhesive under test and the control adhesive. In addition, the anaerobic adhesive was affected by the material composition of the bracket base and curing time. After 1h of curing and using the Miniature Twin bracket, the measured force to debond exceeded the 10 min force to debond results of the control adhesive. SIGNIFICANCE: It is possible to bond commercially available orthodontic brackets to teeth using an anaerobic adhesive. PMID- 16364424 TI - The effect of ytterbium fluoride and barium sulphate nanoparticles on the reactivity and strength of a glass-ionomer cement. AB - OBJECTIVES: While clinical advantages of glass-ionomers include fluoride release and radiopacity, disadvantages include low strength, slow initial setting times and opacity. The addition of nanoparticles, in particular those containing fluoride and cross-linkable ions, may mitigate the disadvantages while further improving the advantages. This investigation evaluated the effects of the addition of ytterbium fluoride (YbF3) and barium sulphate (BaSO4) on the strength and reactivity of a commercial glass-ionomer cement. METHODS: YbF3 and BaSO4 nanoparticles were incorporated into the powder component of Riva SC (SDI Ltd., Bayswater, Australia) at 1, 2, 5, 10, 15, and 25% by weight. Capsules were assembled at a powder:liquid ratio of 2.9:1, activated and mixed, and the resultant pastes evaluated for working time, initial setting time, 24-h surface hardness and 24-h compressive strength. RESULTS: Working and initial setting times were reduced with the addition of YbF3. Addition of BaSO4 at low concentrations reduced working and initial setting times, but further addition delayed the setting reaction. Compressive strength decreased with the addition of either YbF3 or BaSO4, while surface hardness was slightly but insignificantly higher at 1-2% nanoparticles and then decreased with increasing nanoparticle concentrations. SIGNIFICANCE: Nanoparticles modified the setting characteristics, strength and surface hardness of a commercial glass-ionomer cement, and may be useful for refining the handling characteristics of these materials. Further improvements in powder blending may result in more significant improvements in mechanical properties. PMID- 16364425 TI - Non-inverse-square force-distance law for long thin magnets. AB - OBJECTIVE: There is a need for a means of characterizing permanent magnets for dental applications, in particular the force-distance relationship. Problems with determining this on the basis of elementary theory prompted consideration of a model system of long thin magnets. MATERIALS AND METHODS: (a) The force exerted by rod-shaped magnets (Alnico V, 3mm diameter, 170 mm long) perpendicular to a large steel plate, as a function of distance, at small separations (<5mm), was determined for 85 examples. (b) Numerical modelling for a "polar disc" or sheaf of dipoles was conducted to test an alternative physical representation to the normally assumed simple dipole. (c) Exploratory curve-fitting trials were undertaken on the basis of known boundary conditions and observation of actual behaviour. RESULTS: The inverse square law for a point-like pole was found not to apply to the experimental data in any region. The polar disc model was found to provide a very good fit to the experimental data over the whole range tested, but only when an offset ( approximately 1mm) of the pole position from the magnet face was assumed, and that this offset experienced an exponential-decay pole position relaxation (typically 0.4-1.5mm). The fitted pole strength was of the order of 0.18 mAm. SIGNIFICANCE: The implications of these results are that the commonly used elementary view of a simple dipole magnet is of little value for understanding the force-distance relationship and cannot be used as a basis for dental device characterization. PMID- 16364426 TI - Dynamics of composite polymerization mediates the development of cuspal strain. AB - OBJECTIVES: In the current study, we used electronic speckle pattern interferometry (ESPI) to measure tooth deformation in response to polymerization of five resin composites with a range of polymerization shrinkage. Our hypothesis was that composites with higher polymerization shrinkage should cause more cuspal strain as measured by ESPI. METHODS: Standardized MOD cavities were prepared and placed into the ESPI apparatus before the cavities were filled with composites (n=10). The ESPI apparatus was constructed to measure the out-of-plane displacement of the lingual cusps of the teeth during the polymerization of the restorative material. A thermocouple was attached to the specimen to monitor thermal changes throughout the polymerization process. RESULTS: Experiments with empty preparations demonstrated that the ESPI technique was temporally responsive and sensitive to dimensional changes. However, the correlation between polymerization shrinkage of composite resins and ESPI-measured tooth deformation was not straightforward. In particular, a flowable material did not deform the tooth significantly more that a conventional hybrid. Further, an experimental silorane material (with the lowest axial shrinkage) induced the least tooth deformation. SIGNIFICANCE: We concluded that ESPI is a viable method for assessing cuspal strain induced by shrinkage of bonded composite restorations, but that polymerization shrinkage data may overestimate shrinkage-induced tooth deformation. The rate of polymerization shrinkage appeared to mediate the development of cuspal strain. PMID- 16364427 TI - Chemical analysis and bonding reaction of RelyX Unicem and Bifix composites--a comparative study. AB - OBJECTIVES: The chemical and physical properties of the dual curing luting composites RelyX Unicem (3M ESPE) and Bifix (VOCO) were analyzed with regard to their elemental composition, surface morphology and polymerization reaction. The bonding of both materials to hydroxyapatite (HAp) was studied. METHODS: The main components were analyzed by XPS and EDX. The minor components were identified with ICP-OES. Moreover, the morphology was examined by SEM and the polymerization reaction products were investigated using GPC. XPS was also applied to study the bonding mechanisms to HAp. RESULTS: The inorganic product particles consist of an Al-Si-Na-glass network, which incorporates radiopaque strontium and barium for Bifix and strontium and lanthanum for RelyX Unicem. RelyX Unicem contains about 10% fluoride and 2% Ca(OH)(2), whereas Bifix comprises 2% fluoride. After polymerization, reaction products of 10(5)-10(6)g/mol were identified with RelyX Unicem. Both products contain mono- and oligomeric compounds. The reaction with HAp generates calcium atoms with a reduced binding energy. They act as an electron acceptor and show chemical interaction between the composite and HAp. With RelyX Unicem 86% of the calcium atoms reacted, compared to 65% with Bifix. SIGNIFICANCE: The intense chemical interaction of RelyX Unicem with HAp seems to be relevant to clinical aspects and explains the mechanical product properties. After setting, a polymer was found with RelyX Unicem but only monomeric/oligomeric products were identified at the surface of Bifix. PMID- 16364428 TI - Young, black, and male in foster care: relationship of negative social contextual experiences to factors relevant to mental health service delivery. AB - Among a small, cross-sectional sample of young Black males transitioning from foster care (n=74), this study explored the relationship of their negative social contextual experiences to two factors relevant to the delivery of mental health services to them: cultural mistrust of mental health professionals and attitudes toward seeking professional help. Three domains of young Black male's negative social contextual experiences were measured: proximal negative experiences, distal negative experiences, and negative imagery experiences. Results of multivariate analysis of covariance (MANCOVA) controlling for custody status, counselling status and history, and psychiatric history showed that young Black males reporting a high frequency of negative social contextual experiences reported significantly greater cultural mistrust of mental health professionals and significantly less positive attitudes toward seeking professional help for mental health problems than young Black males reporting a low frequency of negative social contextual experiences. Implications and future research directions are discussed. PMID- 16364429 TI - Biochemical consequences of alginate encapsulation: a NMR study of insulin secreting cells. AB - In this study we explore the biochemical consequences of alginate encapsulation on betaTC3 cells. (13)C NMR spectroscopy and isotopomer analysis were used to investigate the effects of encapsulation on several enzymatic processes associated with the TCA cycle. Our data show statistically significant differences in various enzymatic fluxes related to the TCA cycle and insulin secretion between monolayer and alginate-encapsulated cultures. The principal cause for these effects was the process of trypsinization. Embedding the trypsinized cells in alginate beads did not have a compounded effect on the enzymatic fluxes of entrapped cells. However, an additional small but statistically significant decrease in insulin secretion was measured in encapsulated cells. Finally, differences in either enzymatic fluxes or glucose consumption as a function of bead diameter were not observed. However, differences in T(2), assessed by (1)H NMR microimaging, were observed as a function of bead diameter, suggesting that smaller beads became more organized with time in culture, while larger beads displayed a looser organization. PMID- 16364430 TI - Sustained delivery of growth factors from methylidene malonate 2.1.2-based polymers. AB - The incorporation of growth factors into new methylidene malonate 2.1.2-based biocompatible polymeric blends of oligomers and polymers to improve their stability and controlled release was investigated. Five growth factors were used in this study: FGF2, PDGF, TGF-beta, NGF and GM-CSF. Formulation in poly(methylidene malonate 2.1.2) blends was achieved by a four-step optimized process, using different oligomers/polymers ratios. Once dried, formulations could be subsequently stored at 4 or 20 degrees C or immediately subjected to degradation in conditioned cell culture medium. Toxicity of blends and their degradation products were evaluated in several cell lines with MTT. Bioactivity and biospecificity of the formulated growth factors were investigated using MTT and immunohistochemical staining. Combined ELISA and crystal violet colorimetric assays were performed to analyze growth factors release. Limited toxicities were observed for unloaded poly(methylidene malonate 2.1.2) blends. Once optimized, growth factors formulations did not reveal lower bioactivities or loss of biospecificity. Moreover, a sustained release over a 21-day period with more than 90% of preserved bioactivity was reached. To conclude, dual growth factor delivery was made possible by the mean of poly(methylidene malonate 2.1.2) blends. These studies demonstrate the ability of methylidene malonate 2.1.2-based polymeric blends for the delivery of growth factors. PMID- 16364431 TI - Pattern stability under cell culture conditions--a comparative study of patterning methods based on PLL-g-PEG background passivation. AB - Despite the rapidly increasing number of publications on the fabrication and use of micro-patterns for cell studies, comparatively little is know about the long term stability of such patterns under cell culture conditions. Here, we report on the long-term stability of cellular patterns created by three different patterning techniques: selective molecular assembly patterning, micro-contact printing and molecular assembly patterning by lift-off. We demonstrate that although all three techniques were combined with the same background passivation chemistry based on assembly of a PEG-graft copolymer, there are considerable differences in the long-term stability between the three different pattern types under cell culture conditions. Our results suggest that these differences are not cell-dependent but are due to different (substrate-dependent) interactions between the patterned substrate, the passivating molecule and the serum containing cellular medium. PMID- 16364432 TI - Protein interactions with subcutaneously implanted biosensors. AB - Biofouling of in vivo glucose sensors has been indicated as the primary reason for sensitivity losses observed during the first 24 h after implant [Wisniewski N, Moussy F, Reichert WM. Characterization of implantable biosensor membrane biofouling. Fresen J Anal Chem 2000; 366(6-7): 611-621]. Identification of the biomolecules that contribute to these sensitivity perturbations is the primary objective of the research presented. Active needle-type glucose sensors were implanted in Sprague-Dawley rats for 24h, and then a proteomics approach was used to identify the substances absorbed to the sensors. MALDI-TOF mass spectrometry was the primary tool utilized to identify the biomolecules in sensor leachate samples and species absorbed directly on sensor membranes excised from explanted in vivo sensors. Not surprisingly serum albumin was identified as the primary biomolecule present, however, predominantly as endogenous fragments of the protein. In addition, several other biomolecule fragments, mainly less than 15 kD, were identified. Based on these findings, it is concluded that fragments of larger biomolecules infiltrate the sensor membranes causing diminished glucose diffusivity, thus decreasing in vivo sensitivity. PMID- 16364433 TI - Fabrication of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) microstructures using soft lithography for scaffold applications. AB - This paper reports two soft lithographic methods, micromolding and hot embossing, to produce biodegradable poly (3-hydroxybutyrate-co-3-ftydroxyhexanoate) (PHBHHx) arrays of microstructures for hosting and culturing cells in a local microenvironment by controlled shape. Silicon masters with high-aspect-ratio microfeatures were fabricated using KOH and DRIE anisotropic etching. These silicon masters were used as molds to construct PHBHHx microstructures using micromolding and hot embossing. Using silicon rather than conventional PDMS as molds allowed microstructures with feature size of 20 microm and height of 100 microm to be realized. PHBHHx microstructures with different configurations including circles, rectangles, and octagons were fabricated to investigate the effects of topography on cell culture. Mouse fibroblast cell lines L929 were cultured on PHBHHx microstructures in vitro to investigate the biocompatibility. This study demonstrates the feasibility of microfabrication of PHBHHx structures with micro-scale feature size using soft lithography, and the results show that PHBHHx microstructures can be created to mimic cellular microenvironment for cell culture, providing a convenient means to investigate relationships of microstructures and cell functions. PMID- 16364434 TI - The significance of infection related to orthopedic devices and issues of antibiotic resistance. AB - Over the last 15 years, with the advent of modern standards in the control of sterility within the operating room environment and adequate protocols of peri operative antibiotic prophylaxis, the incidence of infections associated to orthopedic implants has become very low. Nevertheless, the event of infection still represents one of the most serious and devastating complications which may involve prosthetic devices. It leads to complex revision procedures and, often, to the failure of the implant and the need for its complete removal. In orthopedics, for the enormous number of surgical procedures involving invasive implant materials, even if nowadays rare, infections have a huge impact in terms of morbidity, mortality, and medical costs. The difficult battle to prevent and fight bacterial infections associated to prosthetic materials must be played on different grounds. A winning strategy requires a clear view of the pathogenesis and the epidemiology of implant-related infections, with a special attention on the alarming phenomenon of antibiotic resistance. In this regard staphylococci are the prevalent and most important causative pathogens involved in orthopedic implant-related infections, and, thus, the main enemy to defeat. In this paper, we offer an overview of the complexity of this battleground and of the current and new, in our opinion most promising, strategies in the field of biomaterials to reduce the risks and counteract the establishment of implant infections. PMID- 16364435 TI - Involvement of calcium mobilization from caffeine-sensitive stores in mechanically induced cell cycle arrest in the dinoflagellate Crypthecodinium cohnii. AB - Mechanical loads can profoundly alter cell growth and cell proliferation. The dinoflagellates are especially sensitive to mechanical stimulation. Many species will be arrested in cell cycle in response to turbulence or shear stress. We demonstrate here that mechanical shaking and caffeine, the ryanodine-receptor agonist, induced an elevation of cytosolic calcium in the dinoflagellate Crypthecodinium cohnii. Dantrolene, a ryanodine-receptor antagonist, dose dependently inhibited both shaking-induced and caffeine-induced calcium release. Similar to the effect of mechanical shaking, caffeine alone dose-dependently and reversibly induced cell cycle arrest in dinoflagellates. Prolonged shaking substantially abolished the magnitude of caffeine-induced calcium release and vice-versa, suggesting that both agents released calcium from similar stores through ryanodine receptors. Fluorescence-conjugated ryanodine gave positive labeling, which could be blocked by ryanodine, in the cortice of C. cohnii cells. In addition, caffeine or shaking mobilized intracellular chlortetracycline (CTC) positive membrane-bound calcium, which could be similarly depleted by t-BuBHQ, a SERCA pump inhibitor. Prior treatment with shaking or caffeine also inhibited the ability of the other agent in mobilizing CTC-positive calcium. CTC-positive microsomal fractions could also be induced to release calcium by caffeine and cADPR, the ryanodinee receptor modulator. t-BuBHQ, but not calcium ionophores, induced cell cycle arrest, and the calcium chelator BAPTA-AM was unable to rescue caffeine-induced cell cycle arrest. These data culminate to suggest that mobilization or depletion of caffeine-sensitive calcium stores, but not calcium elevation per se, is involved in the induction of cell cycle arrest by mechanical stimulation. The present study establishes the role of caffeine-sensitive calcium stores in the regulation of cell cycle progression. PMID- 16364436 TI - Purification, characterization and cDNA cloning of a novel lipopolysaccharide binding lectin from the shrimp Penaeus monodon. AB - In invertebrates, C-type lectin plays an important role in innate immunity by mediating the recognition of pathogens to host cells and clearing microinvaders. A few C-type lectins have been identified from shrimps, but none of their gene or protein sequences is known to date. In this paper, a C-type lectin (named PmLec) specific for bacterial lipopolysaccharide was purified from the serum of the shrimp Penaeus monodon. The binding of PmLec to lipopolysaccharide was mainly mediated through the O-antigen. PmLec had a strong hemagglutinating and bacterial agglutinating activity as well as an opsonic effect that enhances hemocyte phagocytosis. The PmLec cDNA sequence was obtained from the cDNA library of P. monodon by polymerase chain reaction with the degenerated primer designed according to the amino-terminal residue sequence of purified PmLec. A 546-bp open reading frame was found to encode a putative protein comprising 182 amino acids and containing a preceding signal peptide of 17 amino acids. A C-type lectin domain existed in PmLec, but no glycosylation site was found. The recombinant PmLec protein expressed in Escherichia coli also showed the same agglutinating activity and opsonic effect as that of the native protein. This is the first report of a lectin cDNA from the shrimp. PmLec functions as a pattern-recognition protein and an opsonin in the shrimp, and it provides a clue to elucidate the role of lectin in the innate immunity of aquatic invertebrates at the molecular level. PMID- 16364437 TI - A calreticulin-like protein from endoparasitoid venom fluid is involved in host hemocyte inactivation. AB - During oviposition, most endoparasitoid wasps inject maternal factors into their hosts to interfere with host immune reactions and ensure successful development of their progeny. Since encapsulation is a major cellular defensive response of insects against intruding parasites, parasitoids have developed numerous mechanisms to suppress the host encapsulation capability by interfering with every step in the process, including recognition, adherence and spreading. In previous studies, components of Cotesia rubecula venom were shown to inhibit melanization of host hemolymph by interfering with the prophenoloxidase activation cascade and facilitate expression of polydnavirus genes. Here we report the isolation and characterization of another venom protein with similarity to calreticulin. Results indicate that C. rubecula calreticulin (CrCRT) inhibits hemocyte spreading behavior, thus preventing encapsulation of the developing parasitoid. It is possible that the protein might function as an antagonist competing for binding sites with the host hemocyte calreticulin, which mediates early-encapsulation reactions. PMID- 16364438 TI - Characterization of glycans on major histocompatibility complex class II molecules in channel catfish, Ictalurus punctatus. AB - The glycans associated with mammalian major histocompatibility complex (MHC) class II molecules have been studied extensively. Co-translational and post translational addition of sugar molecules to proteins confers many structural and modulatory functions. In the present study we characterized the glycans associated with MHC class II molecules in the channel catfish to compare glycosylation patterns in a teleost to those known to occur in mammals. This study made use of enzymatic methods and two-dimensional (2D) gel electrophoresis to characterize the N-linked sugars. Unlike mammalian T cells which expressed complex N-linked sugars, channel catfish derived 28S T cells were found to express high-mannose/hybrid N-glycans on class II molecules. However studies with Endoglycosidase H in conjunction with cell surface labeling on peripheral blood leukocytes revealed that catfish possess the machinery to modify the intermediate high-mannose sugars to complex type sugars. Nonetheless, the majority of the class II cell surface glycoproteins were of the high-mannose type. Resolution of catfish MHC class II molecules by 2D gel analyses revealed multiple bands for class II beta chains whereas class II alpha chains focused as a single spot. Glycosylation in the channel catfish, a premier model system for studying the immune system of teleosts, has significant differences from the glycosylation patterns characterized in mammalian systems, likely with functional implications. PMID- 16364439 TI - Histological changes in the liver of Oreochromis mossambicus (Cichlidae) after exposure to cadmium and zinc. AB - The toxic effects, of two heavy metals, cadmium (Cd) and zinc (Zn), on the histology of the liver of the southern African freshwater fish Oreochromis mossambicus, were investigated. The goal was to identify whether metal concentrations and exposure period influence the degree and nature of histological changes in the liver of exposed fish. Selected fish were exposed to a mixture of 5% concentrations of the LC(50) of cadmium and zinc and to a mixture of 10% concentrations of the LC(50) of cadmium and zinc, over both short- and long-term exposure periods. Similar histological changes occurred in the livers of specimens exposed to both 5% and 10% concentrations, indicating a definite toxic response to both the metal concentrations. These histological changes included hyalinization, hepatocyte vacuolation, cellular swelling, and congestion of blood vessels. The intensity of these histological changes was, however, influenced by the extent of the exposure period. PMID- 16364440 TI - Intracellular co-localization of SPLUNC1 protein with nanobacteria in nasopharyngeal carcinoma epithelia HNE1 cells depended on the bactericidal permeability increasing protein domain. AB - Epithelial surfaces constitute natural immunobarriers against environmental threats. These barriers are brimming with fluids that bind, transport, cleave or degrade bacterial cells and their endotoxic by-products. Saliva and the airway surface-lining fluid (ASL) comprise the important fluid constituents. Short palate, lung and nasal epithelium clone 1 (SPLUNC1) is a potential host defensive protein that is secreted from the submucosal gland to the saliva and nasal lavage fluid. However, its antimicrobial spectrum and antimicrobial mechanism is not clear. Through green fluorescence protein (GFP) mediated subcellular localization experiments in nasopharyngeal carcinoma (NPC) HNE1 cell line, we determined that the intracellular GFP-tagged SPLUNC1 protein binds to a miniscule microorganisms, approximately 50-400nm in size, after the bactericidal permeability increasing protein (BPI) domain was deleted, GFP-tagged truncated SPLUNC1 protein lost its function of binding to the miniscule microorganisms. We verified that these microorganisms are nanobacteria (NB) with a negative staining using transmitted electronic microscope (TEM) and immunofluorescent analysis using an NB-specific antibody. We isolated and cultured the NB from the cultured nasopharyngeal carcinoma epithelia HNE1 cell supernatant. We found that the NB did not absorb the Hoechst stain, even when we extended the staining time to 35min. However, with the time extension the larger sized NB (larger than 300nm) did stain positively. From the biopsy specimen of NPC, we also detected the NB, which can lead to the swelling of mitochondria in the infected host cells. We hypothesize that SPLUNC1 and NB co-localization is due to the GFP-tagged SPLUNC1 protein binding to the lipopolysaccharide (LPS) of the Gram-negative NB, which can play an important role in the host defense of nasopharyngeal epithelium. This research sheds new light on the mechanism of SPLUNC1 involvement in the host upper respiratory tract defense system. PMID- 16364441 TI - Blockade of interleukin-13-mediated cell activation by a novel inhibitory antibody to human IL-13 receptor alpha1. AB - Interleukin-13 (IL-13) is a cytokine with a crucial role in the development of allergic asthma. The IL-13 receptor shares the IL-4Ralpha subunit with the IL-4R system, but contains as a specific component the IL-13Ralpha1 chain. Blocking signal release by IL-13 without affecting IL-4 function is a potentially interesting therapeutical option for the treatment of asthma. Employing genetic immunization, we generated a set of novel monoclonal antibodies to the IL 13Ralpha1 receptor that proved very specific and efficient inhibitors of human IL 13 activity. Receptor binding antibodies were identified by their specific reactivity with both human monocytes and a murine pro-B cell line overexpressing human IL-13Ralpha1 by flow cytometry and cell ELISA. A luciferase reporter cell system based on STAT6-mediated promoter activation in murine Ba/F3 cells was employed to screen the antibodies for IL-13 antagonistic properties. Inhibitory antibody effects were quantified by interference with IL-13-dependent proliferation of TF-1 cells. The capability of blocking IL-13-driven responses of primary, inflammation-relevant cells was tested by Western blot analysis of STAT6 tyrosine phosphorylation and expression of 15-lipoxygenase in monocytes from fresh blood. The most potent inhibitory antibody identified, GM1E7, inhibited IL 13-driven gene activation and cell proliferation in immune cell lines with IC(50) values in the low nanomolar range. Both short-term (STAT6 activation) and long term (15-LO induction) responses of primary human blood cells to IL-13 were almost entirely blocked, whereas IL-4 effects remained virtually unaffected. GM1E7 is superior to available agents interfering with IL-13 activity in terms of specificity and efficiency and offers potential novel therapeutic perspectives for the treatment of allergic asthma. PMID- 16364442 TI - Copper deficiency increases fibulin-5 (DANCE/EVEC) but decreases cytochrome C oxidase VIb subunit expression in rat heart. AB - It has been well documented that dietary copper (Cu) deficiency causes a hypertrophic cardiomyopathy in rodent models. However, a possible alteration in gene expression has not been fully examined. The present study was undertaken to determine the effect of Cu deficiency on protein profiles in rat heart tissue with the combination of the isotope-coded affinity tag (ICAT) method and Western blotting analysis. Male Sprague-Dawley rats were fed diets that were either Cu adequate (6.0 microg Cu/g diet n=6) or Cu-deficient (0.3 microg Cu/g diet n=6) for 5 week. The ICAT analysis suggested that high-salt buffer (HSB) protein profiles from heart tissue of Cu-deficient rats were different from those of Cu adequate rats; seven major protein species differed by more than a 100% increase or a 50% decrease. With three available antibodies, our Western blotting analysis confirmed that there was an 85% increase in fibulin-5 (also known DANCE/EVEC) and a 71% decrease in cytochrome C oxidase (CCO) VIb subunit, but no change in succinate dehydrogenase complex (also known complex II) IP subunit in Cu deficient rat heart. Collectively, these data may be useful in deciphering the molecular basis for the impairments of function related to the hypertrophic cardiomyopathy of Cu-deficient rats. PMID- 16364443 TI - Synthesis, physicochemical and pharmacokinetic characterization of calcium uronates. AB - Four calcium compounds containing uronic acids (D(+)-galacturonic and D(+) glucuronic) in L:M ratio = 2 and 3 were isolated by applying novel (except for one complex) synthetic procedures. The compounds were characterized by elemental analysis, spectroscopic methods (diffuse reflectance and absorption UV-visible, IR, FIR), mass spectrometry, fast atom bombardment (FAB), thermal decomposition, thermogravimetry/derivative thermogravimetry (TG/DTG) data and differential scanning calorimetric studies (DSC). Two modes of water binding in the complexes, i.e., hydration and coordination-like, were established. Computer-aided analysis has shown that further investigations are needed in order to determine the applicability of calcium uronates as calcium carriers. PMID- 16364444 TI - Invasive group A streptococcal infection with pancarditis caused by a new emm type 12 allele of Streptococcus pyogenes. AB - Throughout the 1980s, a progressive increase in the incidence of Streptococcus pyogenes-related invasive infections has occurred. It has been suggested that a host-related immunogenetic background, as well as bacterial virulence factors may play an important role in the outcome of streptococcal infections. Here, we present the first case of pancarditis in the literature caused by direct bacterial invasion due to a new emm-type 12 allele of S. pyogenes in an immunocompetent patient. The pathogenesis of this invasive infection, as well as predictors of poor prognosis are discussed. PMID- 16364445 TI - Genetically modified plants and food hypersensitivity diseases: usage and implications of experimental models for risk assessment. AB - The recent advances in biotechnology in the plant industry have led to increasing crop production and yield that in turn has increased the usage of genetically modified (GM) food in the human food chain. The usage of GM foods for human consumption has raised a number of fundamental questions including the ability of GM foods to elicit potentially harmful immunological responses, including allergic hypersensitivity. To assess the safety of foods derived from GM plants including allergenic potential, the US FDA, Food and Agriculture Organization of the United Nations (FAO)/World Health Organization (WHO), and the EU have developed approaches for evaluation assessment. One assessment approach that has been a very active area of research and debate is the development and usage of animal models to assess the potential allergenicity of GM foods. A number of specific animal models employing rodents, pigs, and dogs have been developed for allergenicity assessment. However, validation of these models is needed and consideration of the criteria for an appropriate animal model for the assessment of allergenicity in GM plants is required. We have recently employed a BALB/c mouse model to assess the potential allergenicity of GM plants. We have been able to demonstrate that this model is able to detect differences in antigenicity and identify aspects of protein post-translational modifications that can alter antigenicity. Furthermore, this model has also enabled us to examine the usage of GM plants as a therapeutic approach for the treatment of allergic diseases. This review discusses the current approaches to assess the allergenic potential of GM food and particularly focusing on the usage of animal models to determine the potential allergenicity of GM foods and gives an overview of our recent findings and implications of these studies. PMID- 16364446 TI - Cannabinoids, opioids and eating behavior: the molecular face of hedonism? AB - Obesity represents nowadays one of the most devastating health threats. Published reports even project a decline in life expectancy of US citizens due to the rapidly increasing prevalence of obesity. This alarming increase is intimately linked with recent changes of environment and lifestyle in western countries. In this context, the rewarding or even addictive properties of popular food may represent one of the most serious obstacles to overcome for an effective anti obesity therapy. Therefore, in addition to molecular networks controlling energy homeostasis, now researchers are starting to define central nervous mechanisms governing hedonic and addictive components of food intake. A recently emerging body of data suggests that the endogenous cannabinoid and opioid systems both represent key circuits responding to the rewarding value of food. This review focuses on the role of these two systems for the homeostatic and hedonic aspects of eating behavior and includes their anatomical and functional interactions. Independent from the degree to which eating can be considered an addiction, cannabinoid and opioid receptor antagonists are promising anti-obesity drugs, since they are targeting both hedonic and homeostatic components of energy balance control. PMID- 16364447 TI - Two analytical methods to study the interaction of AGEs with cell surface proteins. AB - Advanced glycation end products (AGEs) are sugar-modified proteins that are known to appear in vivo and are suspected to be involved in the pathogenesis of several diseases. Although different cellular responses to AGEs can be measured in cell culture studies, knowledge about the nature of AGE-binding and their cell surface receptors is poor. In the present paper a method for the purification of AGE binding proteins from membrane fractions derived from different rat organs as well as a method for assaying the binding of fluorescein labelled AGEs to the surface of cells of different cell lines are described. The presence of more than 10 proteins interacting with AGEs could be shown in membrane fractions obtained from rat organs. Additionally, binding of AGE-modified BSA to different cells could be shown using fluorescence-labelled ligands in a flow cytometric approach. The presented methods provide an option to isolate AGE-interacting proteins which is a precondition for the identification of these proteins. Furthermore, the measurement of AGE-binding to cell surfaces bears the potential to gain a deeper understanding about the nature of AGE-binding to cell surface proteins and might be applied as a preliminary test before performing cell culture studies about AGE effects. PMID- 16364448 TI - Probing the limits of delay intolerance: preliminary young adult data from the Delay Frustration Task (DeFT). AB - Delay intolerance/aversion is one amongst a number of candidate neuropsychological endophenotypes for ADHD. Pilot data suggest that, because of potential ceiling effects, simple choice measures of delay tolerance used for children are probably not appropriate for adolescents and adults. The Delay Frustration Task (DeFT) is a new measure of delay intolerance, designed to be used in a similar form with adolescents and adults as well as children. In it delay frustration is indexed as the number and duration of responses made on a response key during a series of unpredictable and unsignalled delay periods, which interrupt the completion of a simple computer-based tests. The aim of this study was to provide preliminary data on the applicability of the task in a sample of young adults. The DeFT was administered to 49 male and female undergraduate students selected from a normal population-base. Their mean age was 23.14 (S.D.=1.54). Three measures of delay frustration were recorded across time intervals during the response window; the number of responses, their duration and their combined product (total time button was pressed) was calculated for each second interval bin during the post-response delay period. The AARS and HADS were used as screening questionnaires for ADHD and anxiety behaviour, respectively. The results indicated that young adults with high-ADHD symptoms scores pressed the button more than those with low ADHD scores during the post-response delay condition. While both groups increased responding across time within intervals this was significantly more marked in the high-ADHD symptom group. These effects became more pronounced when anxiety was controlled. Young adults with high-ADHD symptoms appear to be more sensitive to the imposition of unscheduled and unsignalled delay during a simple maths test. DeFT may provide a useful index of delay tolerance in young adults with ADHD. Future research needs to examine DeFT performance in different age groups and in clinical and non-clinical populations. PMID- 16364449 TI - Computerized image analysis for quantitative neuronal phenotyping in zebrafish. AB - An integrated microscope image analysis pipeline is developed for automatic analysis and quantification of phenotypes in zebrafish with altered expression of Alzheimer's disease (AD)-linked genes. We hypothesize that a slight impairment of neuronal integrity in a large number of zebrafish carrying the mutant genotype can be detected through the computerized image analysis method. Key functionalities of our zebrafish image processing pipeline include quantification of neuron loss in zebrafish embryos due to knockdown of AD-linked genes, automatic detection of defective somites, and quantitative measurement of gene expression levels in zebrafish with altered expression of AD-linked genes or treatment with a chemical compound. These quantitative measurements enable the archival of analyzed results and relevant meta-data. The structured database is organized for statistical analysis and data modeling to better understand neuronal integrity and phenotypic changes of zebrafish under different perturbations. Our results show that the computerized analysis is comparable to manual counting with equivalent accuracy and improved efficacy and consistency. Development of such an automated data analysis pipeline represents a significant step forward to achieve accurate and reproducible quantification of neuronal phenotypes in large scale or high-throughput zebrafish imaging studies. PMID- 16364450 TI - A low-cost solution to measure mouse licking in an electrophysiological setup with a standard analog-to-digital converter. AB - Licking behavior in rodents is widely used to determine fluid consumption in various behavioral contexts and is a typical example of rhythmic movement controlled by internal pattern-generating mechanisms. The measurement of licking behavior by commercially available instruments is based on either tongue protrusion interrupting a light beam or on an electrical signal generated by the tongue touching a metal spout. We report here that licking behavior can be measured with high temporal precision by simply connecting a metal sipper tube to the input of a standard analog/digital (A/D) converter and connecting the animal to ground (via a metal cage floor). The signal produced by a single lick consists of a 100-800 mV dc voltage step, which reflects the metal-to-water junction potential and persists for the duration of the tongue-spout contact. This method does not produce any significant electrical artifacts and can be combined with electrophysiological measurements of single unit activity from neurons involved in the control of the licking behavior. PMID- 16364451 TI - Pattern of impaired working memory during major depression. AB - BACKGROUND: The aim of this study was to assess working memory (WM) in patients with major depressive disorder (MDD), using a robust parametric WM task (the n back task). METHODS: Twenty patients with MDD and twenty healthy controls completed a visual version of the paradigm, comprising four levels of task difficulty (i.e. 0-, 1-, 2-, and 3-back). Performance accuracy and reaction time (RT) were measured at each difficulty level. RESULTS: In comparison with controls, patients with MDD exhibited slower RTs (F((1,38)) = 25.16, p < 0.001), and reduced accuracy (F((1,38)) = 5.93, p < 0.001). There was no diagnosis specific effect of task difficulty on performance accuracy. However, the faster response to memory (1-3-back) than to shadowing (0-back) tasks observed in controls was not as pronounced in patients. CONCLUSIONS: These observations support a relatively specific impairment of WM/central executive function in MDD, which may potentially mediate the diverse pattern of cognitive dysfunction noted in MDD. The parametric n-back task is applicable to subjects with MDD and yields results interpretable across the dimensions of task difficulty and performance in controls and patients. PMID- 16364452 TI - Psychiatric comorbidity including nicotine dependence among individuals with eating disorder criteria in an adult general population sample. AB - The goal was to analyze psychiatric disorders among individuals who satisfied at least one eating disorder criterion (EDC). The data derived from a cross sectional survey study with a probability sample of residents of a northern German area with 4075 study participants, aged 18-64 years (participation rate 70.2%). Face-to-face in-home computer-aided interviews (Composite International Diagnostic Interview) were used to assess the diagnostic criteria of eating disorders and nicotine dependence, alcohol dependence, depressive, anxiety, and somatoform disorders according to the Diagnostic and Statistical Manual (DSM-IV). Former or current smokers had an odds ratio (OR) of 1.7 (95% confidence intervals, CI, 1.1-2.4 and 1.1-2.6, respectively) for one EDC compared with never smokers. Subjects with a lifetime psychiatric disorder were more likely to have two or more EDC than individuals who never had the respective disorder (nicotine dependence: OR 2.5, CI 1.5-4.2; alcohol dependence or abuse: OR 2.4, CI 1.2-4.7; depressive disorders: OR 2.2, CI 1.4-3.4; anxiety disorders: OR 2.9, CI 1.9-4.5). To conclude, nicotine dependence, alcohol dependence or abuse, depressive disorders, and anxiety disorders are related to two or more EDC in this adult general population sample. PMID- 16364453 TI - Cortactin is required for integrin-mediated cell spreading. AB - Cortactin is an SH3 domain-containing protein that contributes to the formation of dynamic cortical actin-associated structures, such as lamellipodia and membrane ruffles. Here we show that expression of either the GFP-tagged N terminal or the C-teminal halves of cortactin inhibits significantly the spreading of COS7 cells on fibronectin. Introducing inactivating point mutation into the SH3 domain of the C-terminal half of cortactin suspends the dominant negative effect of the construct. In addition, a vector-based RNA interference was used to knock-down endogenous level of cortactin in cells. We demonstrate that cortactin deficient cells were not able to spread. These results suggest that cortactin is required for integrin-mediated signalling pathways. PMID- 16364454 TI - Regulation of expression and signalling modulator function of mammalian tribbles is cell-type specific. AB - The constant need to respond to changes in the environment is a common feature for all life forms. During evolution, a number of intracellular signal processing systems have evolved to fulfill this requirement. One of the most ancient such systems is the mitogen activated protein kinase (MAPK) signalling network, shared by all eukaryotes. Activation of MAPKs is key to regulation of mitosis and in cellular responses to stress or hormones, for instance. In addition, activity of this signalling system is essential during embryonic development. However, many aspects of MAPK mediated responses are strongly cell-type specific. A family of proteins, called tribbles have recently been described as novel regulators of MAPK function. Our group has previously shown that alterations in tribbles levels lead to profound changes in the activation of the various MAPKs. However, little is known about the cell-type specific aspects of regulation of tribbles expression. Here, we report that expression of all three members of the human tribbles family is dynamically controlled in response to inflammatory stimulation. This regulation, however, is strongly cell-type dependent. Our observations suggest regulation of tribbles expression may play an important role in the cell-type specific cellular responses, mediated by the MAPK network. PMID- 16364455 TI - How antibodies act as natural adjuvants. AB - Antibodies can act like adjuvants. They can potently enhance the antibody response, and in the case of IgG and IgE also the T cell response, to the very antigen they are specific for. In this review we will discuss the recent advances made in our understanding of the underlying mechanisms of antibody-mediated feedback enhancement. The immuno-stimulatory properties of IgM, IgG1, IgG2a, IgG2b, IgG3 and IgE will be reviewed in relationship to the complement system and Fc receptors and the physiological relevance will be discussed. PMID- 16364456 TI - Suppression of natural killer cell activity by morphine is mediated by the nucleus accumbens shell. AB - Despite a wealth of data indicating that morphine modulates immune status by acting at mu-opioid receptors in the brain, there is little known about how the opioid system interacts with other neurotransmitter systems to modulate specific immune parameters. The aim of the present study was to investigate whether dopaminergic projections to the nucleus accumbens are involved in morphine induced suppression of splenic natural killer (NK) cell activity. The results indicate that administration of the dopamine D1 antagonist SCH-23390 into the nucleus accumbens shell, but not core, blocked morphine's suppressive effect on NK activity in male Lewis rats. In support of these findings, the effect of morphine was also prevented by intra-accumbens microinfusions of the dopaminergic immunotoxin anti-DAT-saporin. Additionally, administration of the D1 agonist SKF 38393 into the nucleus accumbens shell produced reductions in splenic NK activity comparable to morphine, suggesting a critical role for D1 receptors in the modulation of NK activity. Collectively, these findings demonstrate that dopaminergic inputs to the nucleus accumbens are critically involved in opioid induced immunosuppression and suggest that opioid-induced increases in D1 receptor activation may have adverse consequences on immune status. PMID- 16364457 TI - Relationship between transducer type and low-frequency hearing loss for patients with ventilation tubes. AB - OBJECTIVE: To determine the relationship between the type of transducer used to perform pure-tone audiometry and the appearance of low-frequency hearing loss at 250Hz and 500Hz for patients with ventilation tubes. METHODS: Air conduction thresholds at 250Hz and 500Hz were measured using Telephonics TDH-49 supra-aural headphones and EARTONE 3-A insert earphones for patients with normal ears (N=16) and patients with ventilation tubes (N=114). Tympanometry was performed on each patient prior to audiometric testing. Audiometric test results obtained in normal ears were compared to results for patients with ventilation tubes. For analysis, the ventilation tube patients were separated into two groups, representative of ventilation tube type. RESULTS: Audiometric results obtained using the two transducer types at 250Hz and 500Hz revealed significant differences in threshold for patients with ventilation tubes. Thresholds obtained using insert earphones were generally worse than thresholds obtained using supra-aural headphones for this group. On average, difference in threshold was 14.15dB worse with insert earphones at 250Hz and 9.75dB worse with insert earphones at 500Hz for patients with Sheehy tubes. Average difference in threshold for patients with Donaldson tubes was 13.93dB worse with insert earphones at 250Hz and 8.93dB worse with insert earphones at 500Hz. In addition, thresholds were more variable for patients with ventilation tubes than normal ears at 500Hz. There were no significant differences in threshold for normal ears using both transducers. CONCLUSIONS: When performing pure-tone audiometry, choice of transducer can influence the accurate identification of a low-frequency hearing loss in patients with ventilation tubes. Low-frequency thresholds were generally worse using insert-style earphones to test subjects with tubes, resulting in the apparent identification of a hearing loss. However, with supra-aural headphones, no low frequency hearing loss existed. There were no significant differences in threshold values using either transducer in normal ears. PMID- 16364458 TI - Stressed-out endogenous cannabinoids relieve pain. AB - A variety of physical and psychological stressors induce analgesia by activating descending systems that project from the brain to the spinal cord. This stress induced analgesia (SIA) is mediated by distinct opioid- and non-opioid-dependent mechanisms. New evidence suggests that non-opioid SIA is mediated by two independent endocannabinoids within the midbrain. Furthermore, novel agents that disrupt breakdown of these endocannabinoids enhance non-opioid SIA and pave the way for novel therapies. PMID- 16364459 TI - Analysis of baculovirus aggregates using flow cytometry. AB - Aggregation of viral particles represents a significant problem for baculoviral stock processing and storage. Aggregation may also affect the results of viral particle counting. A method using flow cytometry was previously developed in our lab to measure the concentration of baculovirus particles produced in insect cell cultures. In the present study, the use of the flow cytometry method was extended to the detection of baculovirus aggregates. Flow cytometry analysis of freshly prepared baculovirus stocks, stained with SYBR Green, generally exhibited a single unimodal distribution; while, baculovirus stocks stored at 4 degrees C for a few months exhibited a bimodal distribution of the fluorescent intensity signal. The bimodal distribution was associated with a decrease in the size of the original viral population and an emergence of a new viral population with a high fluorescence intensity. Treatment of these samples with an endonuclease (Benzonase) confirmed that the new population observed in the flow cytometry analysis is not free cellular DNA. Filtration through 0.22 and 0.45 microm membranes of the stored samples prior to flow cytometry analysis confirmed that the high fluorescence intensity population involved particles larger than a single baculovirus. Exposing freshly amplified baculovirus stocks with a unimodal distribution to a pH of 5.3, a condition known to induce aggregation, showed the emergence of a second population with a bimodal distribution. These results suggest that flow cytometry analysis could be used to detect baculovirus aggregates. The aggregates were associated with high fluorescence intensity populations and the mean green fluorescence intensity of these populations could be used as an indicator of the mean aggregate size. PMID- 16364460 TI - Microinjection of ritanserin into the CA1 region of hippocampus improves scopolamine-induced amnesia in adult male rats. AB - The effect of ritanserin (5-HT2 antagonist) on scopolamine (muscarinic cholinergic antagonist)-induced amnesia in Morris water maze (MWM) was investigated. Rats were divided into eight groups and bilaterally cannulated into CA1 region of the hippocampus. One week later, they received repeatedly vehicles (saline, DMSO, saline+DMSO), scopolamine (2 microg/0.5 microl saline/side; 30 min before training), ritanserin (2, 4 and 8 microg/0.5 microl DMSO/side; 20 min before training) and scopolamine (2 microg/0.5 microl; 30 min before ritanserin injection)+ritanserin (4 microg/0.5 microl DMSO) through cannulae each day. Animals were tested for four consecutive days (4 trial/day) in MWM during which the position of hidden platform was unchanged. In the fifth day, the platform was elevated above the water surface in another position to evaluate the function of motor, motivational and visual systems. The results showed a significant increase in escape latencies and traveled distances to find platform in scopolamine treated group as compared to saline group. Ritanserin-treated rats (4 microg/0.5 microl/side) showed a significant decrease in the mentioned parameters as compared to DMSO-treated group. However, scopolamine and ritanserin co administration resulted in a significant decrease in escape latencies and traveled distances as compared to the scopolamine-treated rats. Our findings show that microinjection of ritanserin into the CA1 region of the hippocampus improves the scopolamine-induced amnesia. PMID- 16364461 TI - Selective increase of dark phase water intake in neuropeptide-Y Y2 and Y4 receptor knockout mice. AB - Neuropeptide-Y (NPY) is involved in the regulation of ingestive behaviour and energy homeostasis. Since deletion of the NPY Y2 and Y4 receptor gene increases and decreases food intake, respectively, we examined whether water intake during the light and dark phases is altered in Y2 and Y4 receptor knockout mice. The water consumption of mice staying in their home cages was measured by weighing the water bottles at the beginning and end of the light phase during 4 consecutive days. Control, Y2 and Y4 receptor knockout mice did not differ in their water intake during the light phase. However, during the dark phase Y2 and Y4 receptor knockout mice drank significantly more (46-63%, P<0.05) water than the control mice. The total daily water intake over 24 h was also enhanced. The enhanced water intake during the dark phase was not altered by the beta adrenoceptor antagonist propranolol or the angiotensin AT1 receptor antagonist telmisartan (each injected intraperitoneally at 10 mg/kg). These data indicate that NPY acting via Y2 and Y4 receptors plays a distinctive role in the regulation of nocturnal water consumption. While beta-adrenoceptors and angiotensin AT1 receptors do not seem to be involved, water intake in Y2 and Y4 receptor knockout mice may be enhanced because presynaptic autoinhibition of NPY release and inhibition of orexin neurons in the central nervous system are prevented. PMID- 16364462 TI - Intestinal bioavailability and biotransformation of 3,3',4,4'-tetrachlorobiphenyl (CB 77) in in situ preparations of channel catfish following dietary induction of CYP1A. AB - Previous studies with the catfish in situ perfused intestinal preparation have demonstrated a significant decline in the intestinal bioavailability of a coplanar polychlorinated biphenyl (PCB), 3,3',4,4'-tetrachlorobiphenyl (CB 77)(14C-TCB) dose in animals pre-exposed in vivo to TCB. This response was accompanied by CYP1A induction in the intestine, but little effect upon the oxidative metabolism of the subsequent in situ dose of [14C]-TCB. To ascertain the basis of these responses and the intestine specific contributions, the intestinal bioavailability and metabolism of [14C]-TCB were examined in the in situ intestinal preparation following in vivo exposure to beta-naphthoflavone (BNF; 0, 10 or 50 mg BNF/kg diet for 10 days), BNF was selected as a known inducer of CYP1A and as a compound with a structure unlikely to influence or directly partake in diffusion based TCB concentration gradients. Appreciable amounts of [14C]-TCB molar equivalents (Meq) reached the perfused circulation of the intestinal preparation for all treatments. While BNF pre-exposure elicited induction of CYP1A activities aryl hydrocarbon hydroxylase (AHH) (9.2-12.5-fold) and elicited modest morphological changes (muciparous) in the intestine these changes were not associated with alterations in [14C]-TCB Meq bioavailability. [14C]-TCB metabolism in the intestinal mucosa ranged between 0.54 and 1.27%, for all treatments. As with bioavailability, intestinal metabolism of [14C]-TCB was not significantly influenced in either extent or profile by induction of CYP1A activity as associated with BNF treatment. Four metabolites were found in mucosal sample extracts of which three were tentatively identified as 2-OH-TCB, 4-OH 3,3',4',5-TCB, and 4,4'-diOH-3,3',5,5' tetrachlorobiphenyl. A fourth unknown metabolite presented chromatographic characteristics suggestive of another dihydroxylated metabolite. These data when examined alone and compared to the literature suggest that the intestine may metabolize [14C]-TCB slowly and independent of CYP1A, resulting in somewhat different profiles than published for other organs. In addition, it is likely that previous [14C]-TCB bioavailability findings in the perfused intestine may be based on TCB concentration gradients rather than biotransformation. PMID- 16364463 TI - Entamoeba histolytica up-regulates the Cdc48-like protein, an AAA family member, during the activation of trophozoites with collagen type I and calcium. PMID- 16364464 TI - Transcriptional regulation of human CC chemokine CCL15 gene by NF-kappaB and AP-1 elements in PMA-stimulated U937 monocytoid cells. AB - CCL15 exerts biological effects on a variety of cells, including monocytes. NF kappaB has been reported to be involved in the transcription of the CCL15 gene. In this study, we have identified an AP-1 element located at -76/-65, which appears to regulate the transcription of the CCL15 gene. We also confirmed that the AP-1 factor binds to the element. Specific inhibitors for MAPK pathways and expression of dominant negative MKK4 or JNK1 reduced PMA-induced transcriptional activation of CCL15. Our findings indicate that transcription of the CCL15 gene is regulated by AP-1 and NF-kappaB through MEK and JNK MAPK pathways in monocytoid cells. PMID- 16364465 TI - Rottlerin inhibits stimulated enzymatic secretion and several intracellular signaling transduction pathways in pancreatic acinar cells by a non-PKC-delta dependent mechanism. AB - Protein kinase C-delta (PKC-delta) becomes activated in pancreatic acini in response to cholecystokinin (CCK) and plays a pivotal role in the exocrine pancreatic secretion. Rottlerin, a polyphenolic compound, has been widely used as a potent and specific PKC-delta inhibitor. However, some recent studies showed that rottlerin was not effective in inhibiting PKCdelta activity in vitro and that may display unspecific effects. The aims of this work were to investigate the specificity of rottlerin as an inhibitor of PKC-delta activity in intact cells and to elucidate the biochemical causes of its unspecificity. Preincubation of pancreatic acini with rottlerin (6 microM) inhibited CCK-stimulated translocation, tyrosine phosphorylation (TyrP) and activation of PKC-delta in pancreatic acini in a time-dependent manner. Rottlerin inhibited amylase secretion stimulated by both PKC-dependent pathways (CCK, bombesin, carbachol, TPA) and also by PKC-independent pathways (secretin, VIP, cAMP analogue). CCK stimulation of MAPK activation and p125(FAK) TyrP which are mediated by PKC dependent and -independent pathways were also inhibited by rottlerin. Moreover, rottlerin rapidly depleted ATP content in pancreatic acini in a similar way as the mitochondrial uncouplers CCCP and FCCP. All studied inhibitory effects of rottlerin in pancreatic acini were mimicked by FCCP (agonists-stimulated amylase secretion, p125(FAK) TyrP, MAPK activation and PKC-delta TyrP and translocation). Finally, rottlerin as well as FCCP display a potent inhibitory effect on the activation of other PKC isoforms present in pancreatic acini. Our results suggest that rottlerin effects in pancreatic acini are not due to a specific PKC-delta blockade, but likely due to its negative effect on acini energy resulting in ATP depletion. Therefore, to study the role of PKC-delta in cellular processes using rottlerin it is essential to keep in mind that may deplete ATP levels and inhibit different PKC isoforms. Our results give reasons for a more careful choice of rottlerin for PKC-delta investigation. PMID- 16364466 TI - Co-transfection with the osteogenic protein (OP)-1 gene and the insulin-like growth factor (IGF)-I gene enhanced osteoblastic cell differentiation. AB - Previous studies from this laboratory showed that the action of Osteogenic Protein-1 (OP-1, BMP-7) on osteoblastic cell differentiation could be enhanced by other protein factors, such as Insulin-like Growth Factor (IGF)-I. In the present study, we examined the effects of co-transfection with a combination of the OP-1 and the IGF-I gene on osteoblastic cell differentiation. The results first showed that fetal rat calvaria (FRC) cells transfected with the OP-1 gene under the control of the cytomegalovirus (CMV) promoter showed substantial production of the OP-1 protein. Transfected FRC cells also showed a DNA concentration-dependent increase in alkaline phosphatase (AP) activity, an osteoblastic cell differentiation marker. Von Kossa-positive nodules, a hallmark of bone formation in long-term cultures of bone-derived cells, were also observed in the transfected cells after 26 days in culture, whereas none were observed in control cells. Co-transfection of FRC cells with the combination of the OP-1 and the IGF I gene resulted in a synergistic stimulation of AP activity. The increase was DNA dose-dependent. The current data show that transfection of OP-1 gene into osteoblastic cells stimulates osteoblastic cell differentiation in vitro. The study further demonstrates the feasibility of employing gene transfer of a second gene in combination with an OP-1 vector to synergistically enhance OP-1 activity. PMID- 16364467 TI - Inflammation and virus in dilated cardiomyopathy as indicated by endomyocardial biopsy. AB - Polymerase chain reaction (PCR) testing of "quick-freeze" endomyocardial biopsy tissue for identification of evidence of viral presence is an important new tool in the investigation of unexplained dilated cardiomyopathy patients; and particularly those who are not improving with conventional treatment. Alternate therapeutic measures, including anti-viral treatment and immunosuppressive therapy, offer promise in contributing to left ventricular systolic function improvement. It is important to include PCR testing for a broad spectrum of cardiotropic viruses prior to the application of immunosuppressive therapy. PMID- 16364468 TI - Hypertrophic cardiomyopathy in a patient with Crohn's disease: a case report. AB - Cardiac manifestations of Crohn's disease are rare; the most common is pericarditis. In the present report we briefly describe a 56-year-old man with Crohn's disease who presented to the emergency department due to paroxysmal atrial flutter. A transthoracic echocardiographic study revealed asymmetric left ventricular hypertrophy without outflow pressure gradient. Reviewing the literature, hypertrophic cardiomyopathy has not been previously described in patients with Crohn's disease. PMID- 16364469 TI - Review of 609 patients with rheumatic fever in terms of revised and updated Jones criteria. AB - BACKGROUND: To investigate the findings and prognosis of rheumatic fever (RF) patients seen in the past 20 years and to compare the last two decades. METHODS: The medical records of all RF patients admitted to Gazi University Department of Pediatric Cardiology during 1982-2002 were reviewed. The decade from 1.1.1982 to 31.12.1991 was designated as 1980s and the following decade as the 1990s. RESULTS: Among the 609 RF cases, there was no difference between the two decades regarding mean age, male/female ratio, most of the minor manifestations and findings of the preceding streptococcal infection. As the rate of carditis declined in 1990s, rates of arthritis and chorea increased. Severity of carditis and admissions with reactivation decreased during 1990s. The two decades did not differ regarding mean age, gender ratio, pericarditis rate, number and type of valvular involvement and sequel of carditis cases. Severity of carditis and number of valvular involvement influenced the first-year prognosis. Almost one third of the arthritis cases had monoarthritis in both decades. Atypical cases with small-joint involvement were detected number of which increased during the 1990s. CONCLUSIONS: The two decades do not seem to differ regarding most of the manifestations of RF. More emphasis should be given to atypical cases such as small-joint involvement and monoarthritis and silent carditis. PMID- 16364470 TI - Sudden anabolic steroid abuse-related death in athletes. PMID- 16364471 TI - Is it necessary to add fibrate to statin therapy in the management of dyslipidemia of metabolic syndrome? AB - The available data have suggested a significant association between hypertriglyceridemia and cardiovascular disease. Although atherogenic dyslipidemia in patients with metabolic syndrome is characterized by high triglyceride, low HDL cholesterol and near normal LDL cholesterol levels, lowering LDL cholesterol is the first priority in treating dyslipidemia in patients with metabolic syndrome. The use of statins as monotherapy is still leaving some of these patients with mixed atherogenic dyslipidemia at high risk for coronary events. So, it seems beneficial to add a fibrate to statin therapy in the management of dyslipidemia of metabolic syndrome, especially in patients with inadequately corrected triglyceride levels with statin monotherapy. PMID- 16364472 TI - Painless acute aortic dissection presenting as discrete supravalvular aortic stenosis. PMID- 16364473 TI - Homocysteine is not associated with arterial thickness and stiffness in healthy middle-aged French volunteers. AB - BACKGROUND: To study the association between the total plasma homocysteine (tHcy) concentration and the carotid artery intima-medial wall thickness (IMT), pulse wave velocity (PWV) and the presence of arterial plaques in a French population. METHODS: Cross-sectional analysis of data from 556 male and 559 female middle aged participants (mean (+/-SD) age 59.6+/-4.7 years) provided by an ongoing intervention trial. RESULTS: Mean geometric tHcy concentration was higher for men than for women (10.6 vs. 8.5 micromol/L, p<0.001) and was associated in the expected direction with known determinants. The mean IMT was 0.71+/-0.1 mm for men and 0.69+/-0.1 mm for women (p<0.001), the mean PWV was, respectively, 12.0+/ 2.8 and 10.9+/-2.2 m/sec (p<0.001), and the percentages of subjects with plaques were, respectively, 40.8% and 22.7% (p<0.001). In men only, the age-adjusted mean IMT and PWV increased with an increasing tHcy concentration: the IMT was 0.71 mm in the first tHcy-quartile and 0.73 mm in the fourth tHcy-quartile (p for linear trend=0.03), the PWV values were, respectively, 11.6 and 12.4 m/sec (p for linear trend=0.01). These associations disappeared after adjustment for conventional cardiovascular disease risk factors. CONCLUSION: In this population, the tHcy concentration was not associated with measures of arterial thickness and stiffness. PMID- 16364474 TI - Neuromuscular implications in left ventricular hypertrabeculation/noncompaction. AB - This review focuses on recent advances in the association between left ventricular hypertrabeculation/noncompaction (LVHT), a form of unclassified cardiomyopathy, and neuromuscular disorders (NMD). So far, LVHT has been found in single patients with dystrophinopathy, dystrobrevinopathy, laminopathy, zaspopathy, myotonic dystrophy, infantile glycogenosis type II (Pompe's disease), myoadenylate-deaminase deficiency, mitochondriopathy, Barth syndrome, Friedreich ataxia, and Charcot-Marie-Tooth disease. Most frequently LVHT is found in patients with Barth syndrome and mitochondrial disorders. The prevalence of LVHT in NMD patients is not known. On the contrary, NMD can be detected in up to four fifths of the patients with LVHT. Because LVHT is associated with an increased risk of rhythm abnormalities and heart failure, it is essential to detect LVHT as soon as possible. Because of adequate therapeutic options, all patients with NMD should undergo a comprehensive cardiological examination as soon as their neurological diagnosis is established. In reverse, all patients with LVHT should undergo a comprehensive neurological investigation following the detection of LVHT. PMID- 16364475 TI - The management of acute myocardial infarction in developing countries. AB - Developing countries contribute a major share to the global burden of cardiovascular disease. Acute myocardial infarction (AMI) in particular remains one of the leading causes of death in the developing world as well as in the developed world. While the risk factors, management and outcome of AMI have been extensively studied in the developed world, limited data is available on this subject from developing countries. The current review looks at the prevalence of the classical coronary artery disease risk factors in developing countries and their association with myocardial infarction, as well as the management and outcome of AMI patients in these countries. PMID- 16364476 TI - Aortic elastic properties in athletes using anabolic-androgenic steroids. AB - The use of anabolic-androgenic steroids (AAS) has been linked to acute cardiovascular events in athletes. The purpose of the present study was to investigate the aortic elastic properties in athletes who had been self administering AAS compared with a group of athletes not using these drugs. Fourteen male bodybuilders using AAS and 27 male wrestlers (non-users) volunteered to the study. All subjects were placed in a mild recumbent position and the ascending aorta was recorded in the two-dimensional guided M-mode tracings. Although the aortic distensibility was found to be reduced in user athletes (2.1+/-1.1 vs. 3.8+/-1.4 cm(2) dyn(-1) 10(-6), p=0.01; 9.3+/-3.7 vs. 5.9+/-2.5, p=0.003, respectively). The results of this study indicate that aortic stiffness is increasing in athletes using AAS. PMID- 16364477 TI - Direct comparison of single-strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) to characterize a microbial community on the basis of 16S rRNA gene fragments. AB - Characterization of microbial communities using single-strand conformation polymorphism (SSCP) was compared with that using denaturing gradient gel electrophoresis (DGGE). This comparison was based on the V3-4 region (Escherichia coli positions: 341-806) of 16S rRNA gene of bacterial or archaeal communities obtained from a methanogenic bioreactor. Significant differences in the bacterial banding profiles were observed while attempting to detect the diversity of the community and its succession during the reactor operation. The SSCP produced a number of sharp bands and differentiated the bacterial community structures to which the DGGE gave an identical pattern. On the other hand, the SSCP and DGGE provided similar succession patterns for archaeal community. PMID- 16364478 TI - Evaluation of probe chemistries and platforms to improve the detection limit of real-time PCR. AB - A validated PCR-based Salmonella method targeting a 94-bp sequence of the ttr gene was used as a model to compare six different combinations of reporter and quencher dyes of a TaqMan probe, on three different instruments, to improve the detection limit in a real-time PCR assay with the aim of a same-day analysis. The use of locked nucleic acids (LNA) and Scorpion probes were also tested. The combination FAM-BHQ1 or Cy5-BHQ3, both dark quenchers, gave the best results (Cycle threshold (Ct) of 25.42+/-0.65 and 24.47+/-0.18 at 10(3) DNA copies). When comparing different probe technologies, the LNA probe (FAM-BHQ1) was the most sensitive with the strongest fluorescence signal (dR last 48066), resulting in 0.6 to 1.1 lower Ct values than a DNA TaqMan probe, and 1.9 to 4.0 lower Ct than the Scorpion system (FAM-BHQ1). The RotorGene real-time PCR instrument gave 0.4 1.0 lower Ct values (more sensitive) than the Mx3005p, and 1.5-3.0 lower than the ABI 7700. Using the LNA in a RotorGene instrument, we detected the following Salmonella DNA copies in 1-ml pre-enriched samples: fishmeal (100 copies), chicken rinse (100 copies) and pig feces (10 copies). The detection probability of the final assay on inoculated fecal samples was 100% at 2x10(4) copies per ml. In conclusion, the LNA probe with annealing temperature of 65 degrees C could be useful for more sensitive detection limits. PMID- 16364479 TI - Relationship of P300 single-trial responses with reaction time and preceding stimulus sequence. AB - Variation of single-trial P300 responses was studied both in relation to reaction times and to the preceding stimulus sequence in an auditory oddball paradigm. Single-trial responses were estimated with the Subspace regularization method that is based on Bayesian estimation and linear modeling. The results of the single-trial method were compared to those of averaging. Both methods showed that the latency of the P300 was shorter and its amplitude larger for faster than slower reaction times. The P300 latency was shorter for target tones that were preceded by a large number of standard tones compared to those preceded by a small number of standard tones. The P300 amplitude was statistically significantly affected by the stimulus sequence only when analyzed with conventional averaging. In-depth analysis of standard deviations showed that the variability of the P300 single-trial latencies could explain the differences between the two methods. Specifically, the regression analysis showed that the latency correlated negatively with the number of preceding standard tones and positively with the reaction time, whereas the P300 amplitude correlated positively with the number of the preceding standard stimuli and negatively with the reaction time. The analysis of the single-trial responses gives information about the behavior of the P300 component that is lost with conventional averaging. The method used in this study is independent of subjective decision making and can be used to model changes in the dynamical behavior of the P300 component objectively. PMID- 16364480 TI - Evidence of functional hyperemia in the rat hippocampus during mild treadmill running. AB - Exercise appears to improve hippocampal plasticity and cognitive function, leading us to postulate that exercise may activate hippocampal neurons, which in turn increase hippocampal cerebral blood flow (Hip-CBF). Recent studies using laser-Doppler flowmetry (LDF) have shown that Hip-CBF increases with behavior and locomotion, but it has not been examined whether these changes are due to neuronal activation. We aimed to examine whether functional hyperemia, an increase in cerebral blood flow in response to neuronal activation, can occur in the exercising rat hippocampus. We applied a treadmill running model of rats and LDF combined with microdialysis. Prolonged mild treadmill running (10 m/min) resulted in an increase in Hip-CBF of 26+/-9% versus basal level. When tetrodotoxin was infused via microdialysis into the loci where Hip-CBF was monitored, the increased Hip-CBF with running was completely suppressed. These results provide evidence that functional hyperemia occurs in the rat hippocampus during mild treadmill running and suggest that our running animal model may be useful for examining the underlying mechanisms of exercise-induced hippocampal functional hyperemia. PMID- 16364481 TI - A time course analysis of cyclooxygenase-2 suggests a role in spatial memory retrieval in rats. AB - We previously showed a role for COX-2 in spatial memory retention. In that study we investigated the effects of post-training intrahippocampal infusion of celecoxib as a COX-2-specific inhibitor on spatial memory retention. Those infusions impaired spatial memory retention in the Morris water maze. In the present study a time course analysis of role of COX-2 in spatial memory was conducted. Here stereotaxic surgery was employed for the bilateral implantation of guide cannulas into the CA1 region of the hippocampus. Training trials were started after recovery of the animals. Immediately after last trial of training on third day, the celecoxib (0.1M) was infused bilaterally and testing trials, were performed 1, 2, 3, and 7 days after celecoxib infusions. Significant alterations were observed in escape latency and traveled distance 2 and 3 days after celecoxib infusions. The maximum impairment was obtained 72 h after the infusions. The data suggests that the effect of celecoxib is transient and that its effect on performance is likely caused by a problem in memory retrieval. Quantification analyses of the immunostaining of COX-2-containing neurons in the dorsal hippocampus show that celecoxib infusions significantly reduced (P<0.05) COX-2 immunoreactivity for the animals that were tested 3 days after the drug infusion. Results from the behavioral study along with the findings from immunohistochemical analyses suggest that COX-2 has significant role in spatial memory retrieval. Moreover, the memory deficits induced by the infusions continuously persists for 3 days. PMID- 16364482 TI - Effect of supplementing terpenoid biosynthetic precursors on the accumulation of bilobalide and ginkgolides in Ginkgo biloba cell cultures. AB - The effect of precursor feeding on the production of bilobalide and ginkgolides was studied with suspension cell cultures of Ginkgo biloba. The precursors greatly influenced the productivity of bilobalide and ginkgolides. Precursor supplementation increased the accumulation of both bilobalide and ginkgolides, and with positive effect on cell growth. The GA accumulation by cell cultures was influenced by precursors upstream in the metabolism, whereas the BB accumulation was under the influence of downstream precursors of the terpenoid biosynthetic pathway. Furthermore, precursor feeding modified the ratios of the BB, GA and GB in cells and cell cultures of G. biloba. The studies also aid in understanding effect of precursor feeding on the bilobalide and ginkgolides biosynthetic pathway. PMID- 16364483 TI - Stabilized baculovirus vector expressing a heterologous gene and GP64 from a single bicistronic transcript. AB - The efficient scale-up of recombinant protein production in insect-cell bioreactors using baculovirus expression vectors is hampered by reductions in yield with increasing viral passage, the so-called passage effect. This phenomenon is characterized by the generation and subsequent accumulation of defective interfering baculoviruses (DIs), which interfere with the replication of genomically intact virus. A novel baculovirus expression vector is presented equipped with a bicistronic expression cassette that allows the simultaneous expression of the recombinant gene (GFP, first cistron) and an essential baculovirus gene (GP64, second cistron) from a single messenger RNA (mRNA). The translation of GP64 is mediated by an internal ribosome entry site (IRES) element from Rhopalosiphum padi virus (RhPV) while the native GP64 gene is deleted. In this way, a dominant selection pressure is placed on the entire bicistronic mRNA and hence on the maintenance of the foreign gene. The bicistronic expression vector was superior to the control baculovirus vector in that GFP expression remained at much higher levels upon continued virus passage. The versatility of this stabilized vector was demonstrated by its ability to propagate in a number of cell lines including Sf21, Sf9 and High Five cells. This novel baculovirus vector is especially valuable for large-scale recombinant protein production in insect-cell bioreactors where the number of viral passages is high. PMID- 16364484 TI - A nanocompartment system (Synthosome) designed for biotechnological applications. AB - A nanocompartment system based on two deletion mutants of the large channel protein FhuA (FhuA Delta1-129; FhuA Delta1-160) and an ABA triblock copolymer (PMOXA-PDMS-PMOXA) has been developed for putative biotechnological applications. FhuA is ideally suited for applications in biotechnology due to its monomeric structure, large pore diameter (39-46 A elliptical cross-section) that ensures rapid compound flux, and solved crystallographic structure. Two areas of application were targeted as proof of principle: (A) selective product recovery in nanocompartments and (B) enzymatic conversion in nanocompartments. Selective recovery of negatively charged compounds has been achieved on the example of sulforhodamine B by using positively charged polylysine molecules as trap inside the nanocompartment. Conversion in nanocompartments has been achieved by 3,3',5,5'-tetramethylbenzidine oxidation employing horseradish peroxidase (HRP). PMID- 16364485 TI - Alternative peptide-fusion proteins generated by out-of-frame mutations, just upstream ORFs or elongations in mutants of human hepatitis B viruses. AB - By various means including out-of-frame mutations, just upstream ORFs and elongations, additional peptide fusions could be generated by mutants of Human Hepatitis B Virus (HBV). Numerous frameshift mutations inducing long alternative open reading frames have been evidenced in all HBV genes. Interestingly, these mutants are frequently detected in severe liver diseases, but seldom in asymptomatic carriers. The high level of conservation of some of these sequences in spite of the fact that they could be generated by different types of mutations, as their presence in mutants found on various continents, suggest that these mutations could play a role. These mutants could combine two advantages, that related to the loss of a part of a wild-type protein and that related to the putative advantage conferred by the additional sequences. In addition, in numerous Asian genomes (more than 300 to date) pre-X or pre-pre-S regions were found just upstream to, respectively, the X and the pre-S1 genes. These two regions are translated with their respective genes in frame and recent studies have evidenced the transactivating role of the corresponding proteins. With some exceptions, these regions are genotype- and serotype-specific (C/adr). In addition, these mutants have been found principally in patients with severe hepatitis diseases, for example, hepatocarcinoma in more than one third of the cases. As additional sequences generated by HBV variants may be relevant for viral life cycle, persistence and pathogenesis, further investigations are necessary to give a clearer picture of the subject. PMID- 16364486 TI - [Early prediction of functional outcome one year after initial unilateral hemispheric stroke]. AB - BACKGROUND AND PURPOSE: Several prognostic factors have been identified for outcome after stroke. We conducted a study to determine early predictive factors of functional outcome one year after stroke and to evaluate which factors are independent predictors, with an aim of specifying the role of age, aphasia, unilateral neglect, cognitive impairment and family social support. METHODS: Observational cohort study of 156 patients. All patients admitted to the university hospital for initial unilateral hemispheric stroke were included. The study duration was two years (inclusion, one year, and follow-up, one year) . The initial evaluation of stroke was conducted at day 2 and day 15 and included the Motricity Index and Trunk Control Test, New Functional Ambulation Classification, Frenchay arm test, Mini-Mental State Examination, Boston Diagnostic Aphasia Examination, unilateral neglect evaluation, and depression. Data on functional recovery (Barthel Index) were collected at day 360. RESULTS: The average age of patients was 72 years. Age was correlated to social situation (P<0.01) and previous neurological impairment (P<0.01). A multiple regression analysis, including 14 initial clinical factors correlated with the Barthel Index score at day 360, revealed 4 independent early predictive factors of outcome: initial score of Barthel Index at day 2 and its progression from day 2 to day 15, disorders of the executive functions and previous neurological impairment. CONCLUSION: In our cohort, in accordance with previous studies, age, cognitive impairment, unilateral neglect, aphasia, depression and social situation are not independent factors of poor outcome after stroke as evaluated by the Barthel Index. PMID- 16364487 TI - Hepatotoxicity of antiretrovirals: incidence, mechanisms and management. AB - One of the toxicities linked to the use of antiretrovirals is the elevation of transaminases. Liver toxicity is a cause of morbidity, mortality, and treatment discontinuation in HIV-infected patients. While several antiretrovirals have been reported to cause fatal acute hepatitis, they most often cause asymptomatic elevations of transaminases. Liver toxicity is more frequent among subjects with chronic hepatitis C and/or B. The incidence of drug-induced liver toxicity is not well known for most antiretrovirals. The contribution of each particular drug to the development of hepatotoxicity in a HAART regimen is difficult to determine. Possible pathogenic mechanisms involved in hepatotoxicity are multiple, including direct drug toxicity, immune reconstitution in the presence of HCV and/or HBV co infections, hypersensitivity reactions with liver involvement, and mitochondrial toxicity. Other pathogenic pathways may be involved, such as insulin resistance caused by several antiretrovirals, which may contribute to the development of steatohepatitis. The management of liver toxicity is based mainly on its clinical impact, severity and pathogenic mechanism. PMID- 16364488 TI - Insulin resistance, adiponectin, cytokines in NASH: Which is the best target to treat? PMID- 16364490 TI - Effect of correcting transferrin saturation for body mass index in HFE C282Y homozygotes. PMID- 16364491 TI - The face of future hepatitis C antiviral drug development: recent biological and virologic advances and their translation to drug development and clinical practice. PMID- 16364492 TI - Cellular and virological mechanisms of HBV drug resistance. PMID- 16364493 TI - Bacteriophage origins of mitochondrial replication and transcription proteins. AB - Mounting evidence suggests that key components of the mitochondrial transcription and replication apparatus are derived from the T-odd lineage of bacteriophage rather than from an alpha-Proteobacterium, as the endosymbiont hypothesis would predict. We propose that several mitochondrial replication genes were acquired together from an ancestor of T-odd phage early in the evolution of the eukaryotic cell, at the time of the mitochondrial endosymbiosis. We further propose that at a later stage the single-subunit RNA polymerase, originally acquired for mitochondrial DNA replication, was co-opted to serve in mitochondrial transcription. PMID- 16364494 TI - EKB-569, a new irreversible epidermal growth factor receptor tyrosine kinase inhibitor, with clinical activity in patients with non-small cell lung cancer with acquired resistance to gefitinib. AB - EKB-569 is a potent, low molecular weight, selective, and irreversible inhibitor of epidermal growth factor receptor (EGFR) that is being developed as an anticancer agent. A phase 1, dose-escalation study was conducted in Japanese patients. EKB-569 was administered orally, once daily, in 28-day cycles, to patients with advanced-stage malignancies known to overexpress EGFR. Two patients with advanced non-small cell lung cancer with EGFR mutations and acquired gefitinib resistance from the phase 1 study are described in detail. Case #1 is a 63-year-old man with smoking history. He received treatment from 4 March 2004. Because he had no severe adverse events, a total of 10 courses of therapy were completed through December 16. Grade 2 skin rash and ALT elevation, and grade 1 diarrhea and nail changes developed. A chest CT scan on 4 August 2003 revealed multiple pulmonary metastases that had decreased in size. Case #2 is a 49-year old woman with no smoking history. She received therapy from 9 February 2004. She received a total of five courses of the therapy until 22 June 2004. Grade 3 nausea and vomiting and grade 1 diarrhea and dry skin developed. A chest CT scan on March 3 revealed multiple pulmonary metastases that had decreased in size. A brain MRI on March 4 showed that multiple brain metastases also had decreased in size. Based on RECIST criteria, they had stable disease but radiographic tumor regression was observed. PMID- 16364495 TI - The effect of silica on the degradation of organohalides in granular iron columns. AB - Dissolved silica species are naturally occurring, ubiquitous groundwater constituents with corrosion-inhibiting properties. Their influence on the performance and longevity of iron-based permeable reactive barriers for treatment of organohalides was investigated through long-term column studies using Connelly iron as the reactive medium. Addition of dissolved silica (0.5 mM) to the column feed solution led to a reduction in iron reactivity of 65% for trichloroethylene (TCE), 74% for 1,1,2-trichloroethane (1,1,2-TCA), and 93% for 1,1,1 trichloroethane (1,1,1-TCA), compared to columns operated under silica-free conditions. Even though silica adsorption was a gradual process, the inhibitory effect was evident within the first week, with subsequent decreases in reactivity over 288 days being relatively minor. Lower concentrations of dissolved silica species (0.2 mM) led to a lesser decrease (70%) in iron reactivity toward 1,1,1 TCA. The presence of dissolved silica species produced a shift in TCE product distribution toward the more highly chlorinated product cis-dichloroethylene (cis DCE), although it did not appear to alter products originating from the trichloroethanes. The major corrosion products identified were magnetite (Fe3O4) or maghemite (gamma-Fe2O3) and carbonate green rust ([Fe4(2+)Fe(2)3+(OH)12][CO(3).2H2O]). Iron carbonate hydroxide (Fe(II)1.8Fe(III)0.2(OH)2.2CO3) was only found in the silica-free column, indicating that silica may hinder its formation. A comparison with columns operated under the same conditions, but using Master Builder iron as the reactive matrix, showed that Connelly iron is initially less reactive, but performs better than Master Builder iron over 288 days. PMID- 16364496 TI - Effects of consuming mycoprotein, tofu or chicken upon subsequent eating behaviour, hunger and safety. AB - This study tested if: (1) a preload of mycoprotein and tofu consumed before a lunch meal have a greater effect on satiety when compared to a chicken preload, (2) the mycoprotein and tofu preloads, compared to chicken, are not associated with compensation or eating more food at a subsequent dinner meal. These hypotheses were tested in a controlled laboratory study using universal eating monitors to measure food intake and visual analogue scales to monitor hunger and satiety. Forty-two overweight adult females consumed three meals in the laboratory on 3 test days. At lunch, isocaloric pasta preloads, containing mycoprotein, tofu, or chicken, varied across the days in a balanced order. The findings of the study supported the two hypotheses. Mycoprotein and tofu preloads, in comparison to the chicken preload, were associated with lower food intake shortly after consuming the preload at lunch. Food intake following consumption of mycoprotein and tofu did not differ, and participants did not compensate for lower food intake at lunch by consuming more food at dinner. The findings suggest that mycoprotein and tofu have satiating properties that persist for several hours after a meal. These findings have significant implications for the development of foods that are low in kilojoules, but are also filling. PMID- 16364497 TI - Effects of hunger and visuo-spatial interference on imagery-induced food cravings. AB - The present study investigated the effects of hunger and visuo-spatial interference on imagery-induced food cravings. Forty-two women were randomly assigned to a hungry (no food for prior 4h) or not hungry condition. Participants were asked to form and maintain images of desired foods while looking at a blank computer screen (control condition) or performing a task designed to load the visuo-spatial sketchpad of working memory (dynamic visual noise). They then rated the vividness of their images and their craving intensity. Although hungry participants reported stronger food cravings, dynamic visual noise made images less vivid and cravings less intense, irrespective of participant hunger status. Thus concurrent visuo-spatial processing may offer a useful technique for treating problematic food cravings that are predominantly psychological in origin, as well as those that are hunger-driven. PMID- 16364498 TI - Responsivity to food stimuli in obese and lean binge eaters using functional MRI. AB - Functional neuroimaging was employed to study 10 obese and 10 lean healthy young right-handed women, divided equally into binge and non-binge eaters. Subjects were presented with visual and auditory stimuli of binge type foods, non-binge type foods, and non-food stimuli in the fMRI scanner. Brain areas activated by both the visual and auditory stimuli across all individual subjects within a particular group was observed only for the binge food stimuli in the obese binge eaters, in the right premotor area, involved in planning of motor behavior. For four of the five obese binge eaters, the activation was in the ventral premotor cortex adjacent to the oral region, and may reflect past or concurrent motor planning about eating binge foods. Because a random effects group analysis has not yet been completed, this should be considered a preliminary report. PMID- 16364499 TI - Circulating urotensin II levels in moderate to severe congestive heart failure: its relations with myocardial function and well established neurohormonal markers. AB - Urotensin II (UII) is a potent vasoactive cyclic peptide thought to play a role in myocardial hypertrophy and remodelling. We therefore determined UII plasma levels in congestive heart failure (CHF) patients and its relationship with the severity of the disease and well-established markers of left ventricular function. UII was significantly higher in CHF patients (n = 57) than in controls (n = 48) [geometric mean (pg/ml), 95% PI: 1.32 (0.67-2.59) versus 0.84 (0.31 1.61), p < 0.0001], was related to the functional class of the disease and correlated negatively with left ventricular ejection fraction (r = -0.316, P = 0.016). Furthermore, UII correlated significantly with Big-ET1 (r = 0.32, p = 0.03), BNP (r = 0.42, p = 0.005) but poorly with Nt-proANP (r = 0.28, p = 0.07). Our results suggest that UII could play a role in worsening the course of congestive heart failure and is associated with established markers of cardiovascular dysfunction. PMID- 16364500 TI - Tissue distribution and plasma clearance of heparin-binding EGF-like growth factor (HB-EGF) in adult and newborn rats. AB - Heparin-binding EGF-like growth factor (HB-EGF), a member of the epidermal growth factor (EGF) family, can protect intestinal epithelial cells from various forms of injury in vitro and attenuate intestinal ischemia/reperfusion damage in vivo. With the goal of eventual clinical use of HB-EGF to protect the intestines from injury in neonates, children, and adults, the pharmacokinetics and biodistribution of 125I-labeled HB-EGF were investigated. After intravenous bolus, HB-EGF had a distribution half-life of 0.8 min and an elimination half life of 26.67 min. After gastric administration, the bioavailability was 7.8%, with a 2.38 h half-life in the absorption phase and an 11.13 h half-life in the elimination phase. After intravenous dosing, most radioactivity was found in the plasma, liver, kidneys, bile, and urine, whereas it was mainly distributed in the gastrointestinal tract after intragastric administration. The degradation of 125I HB-EGF in plasma from newborn rats was lower than that in adult rats after gastric administration. This supports the feasibility of enteral administration of HB-EGF in the treatment of gastrointestinal diseases, including newborns afflicted with necrotizing enterocolitis. PMID- 16364502 TI - Effects of Alzheimer's peptide and alpha1-antichymotrypsin on astrocyte gene expression. AB - We employed gene array technology to investigate the effects of alpha1 antichymotrypsin (ACT), soluble or fibrillar Alzheimer's peptide (Abeta(1-42)) alone and the combination of ACT/Abeta(1-42) on human astrocytes. Using a 1.2 fold change as significance threshold, 398 astrocyte genes showed altered expression in response to these treatments compared to controls. Of the 276 genes affected by the ACT/soluble Abeta(1-42) combination, 195 (70.6%) were suppressed. The ACT/fibrillar Abeta(1-42) combination affected expression of 64 genes of which 58 (90.5%) were up-regulated. The most prominent gene expression changes in response to the ACT/soluble Abeta(1-42), were the down-regulation of at least 60 genes involved in transcription, signal transduction, apoptosis and neurogenesis. The ACT/fibril Abeta(1-42) increased the expression of genes involved in transcription regulation and signal transduction. Surprisingly, gene expression of astrocytes exposed to soluble or fibrillar Abeta(1-42) alone was largely unaffected. Thus, the molecular forms generated by the combination of ACT/Abeta(1 42) alter expression of astrocyte genes more profoundly in breadth and magnitude than soluble or fibrillar Abeta(1-42) alone, suggesting that pathogenic effects of Abeta(1-42) may occur as a consequence of its association with other proteins. PMID- 16364503 TI - [Cryptococcal infections in non-HIV infected patients. Study of four cases and review of literature]. AB - BACKGROUND: Cryptococcal infections are frequent in HIV-infected patients and are regularly looked after. This infection may occur in others immunosuppressives situations and, in those cases, diagnosis is often delayed. METHODS: We report four cases of cryptococcal infections in patients whose immunosuppression isn't related with HIV infection but due to chronic lymphocytic leukemia, giant cell temporal arteritis, gastric neoplasm and lupus. Diagnosis, prognostic and treatment are detailed. RESULTS: Four patients aged from 25 to 76 presented a cryptococcal infection (three meningitis). A woman died at the admission. Another died seven years later. The two others are still alive under treatment. When infected, all patients were immunodeficiency. CONCLUSION: Cryptococcal infection may occur in patients non-HIV-infected patients. Early detection is needed to improve prognostic. PMID- 16364504 TI - [Nitrofurantoin-induced lupus-like syndrome associated with hepatitis]. PMID- 16364505 TI - [An unusual diagnostic of quadriparesia: hyperkalemic paralysis. Report of four non-familial cases]. AB - INTRODUCTION: The classical cause of hyperkalemic paralysis is the hereditary hyperkalemic paralysis. Very rarely, secondary forms of hyperkalemic paralysis have been reported. EXEGESIS: Four cases of acute paralysis mimicking Guillain Barre syndrome in three cases and revealing severe hyperkalemia are presented. All the four patients had moderate chronic renal insufficiency. In two cases, the patients received spironolactone. One case was associated with lysis syndrome. All the 4 cases dramatically improved with the treatment of hyperkalemia. CONCLUSION: These cases pointed out the possibility for acute peripheral paralysis to reveal severe hyperkalemia. PMID- 16364506 TI - [Calcific uremic arteriolopathy (calciphylaxis): an independent vascular risk factor?]. PMID- 16364507 TI - [A possible complication of sickle-cell disease: pulmonary aspergillosis]. PMID- 16364508 TI - [Preeclampsia and antiphospholipid syndrome. Uterine artery Doppler evaluation]. AB - INTRODUCTION: Women with persistently high resistance in uterine arteries have an increased risk of the subsequent development of preeclampsia. Doppler investigation provides a non-invasive method for the study of the uteroplacental blood flow. In pregnant women the antiphospholipid syndrome is associated to an increased risk of preeclampsia and complications related to uteroplacental insufficiency, and the role of uterine artery Doppler is discussed. CURRENT KNOWLEDGE AND KEY POINTS: In normal pregnancy, impedance to flow in the uterine arteries decreases with gestation, as the likely consequence of the physiologic change of spiral arteries into low-resistance vessels. In women with preeclampsia or related complications, the abnormal persistence of high resistance to flow in the uterine arteries correlates with maternal and neonatal outcome. In one study including patients with antiphospholipid syndrome, a high resistance index in the uterine arteries at 22-24 weeks gestation strongly predicted the subsequent development of preeclampsia. In another study including patients with lupus anticoagulant, persistent bilateral notches at 22-24 weeks gestation may identify preeclampsia and fetal growth restriction with a high sensitivity, specificity, positive and negative predictive value. The treatment may improve the uteroplacental blood flow and is a possible confounding factor which needs further evaluation. FUTURE PROSPECTS AND PROJECTS; In patients with antiphospholipid antibodies a higher impedance has been observed in the uterine artery, suggesting a possible vascular dysfunction precluding to impaired trophoblastic invasion and placental thrombosis, as probable mechanisms in the complications. If confirmed, these findings might have important implications for the management of these patients. PMID- 16364509 TI - [Deafness, diabetes and cancer: chance association or etiological link?]. PMID- 16364510 TI - A comparison of problem-based learning and lecture-based learning in an adult health nursing course. AB - PURPOSE: This study aimed to compare the effects of the problem-based learning (PBL) method with the traditional lecture method on learning in the cardiorespiratory nursing section of the Adult Health Nursing course. METHOD: A pretest-posttest experimental design was used. A total of 71 second-year nursing students in a three-year nursing program in Korea participated: 35 students in the PBL group in the fall semester of 2002, and 36 students in the traditional lecture group in 2003. The seven PBL packages were developed by the authors, based on an analysis of relevant learning content and clinical scenarios. RESULTS: The level of knowledge in the PBL group was significantly higher than that of students in the lecture group (t=2.007, p=.045). All PBL students with higher and lower grades showed a significant increase in the posttest score. But in the lecture group, only students with higher grades showed a notable increase. No statistically significant difference was found between the PBL and lecture groups in the level of attitude toward learning (t=1.669, p=.100). Learning motivation was significantly higher in the PBL group (t=2.608, p=.012). CONCLUSION: Students in the PBL group gained more knowledge and had higher motivation toward learning compared to students in the lecture group. PMID- 16364511 TI - Introducing point and deletion mutations into the P/C gene of human parainfluenza virus type 1 (HPIV1) by reverse genetics generates attenuated and efficacious vaccine candidates. AB - The P/C gene of human parainfluenza virus type 1 (HPIV1) encodes a nested set of related accessory C proteins, C'/C/Y1/Y2, which have been shown in other paramyxoviruses to have a role in evasion of the type I interferon (IFN) response following virus infection. We previously demonstrated that a set of two amino acid substitutions, CR84G/HNT553A, and a separate amino acid substitution, CF170S, are independently attenuating for HPIV1 in African green monkeys (AGMs). However, in each case the attenuation (att) phenotype is vulnerable to reversion by a single nucleotide change back to wild type. Using reverse genetics, recombinant HPIV1 (rHPIV1) vaccine candidates were generated that were designed for increased genetic and phenotypic stability by: (i) creating a two-amino acid deletion and substitution at the site of the CF170S mutation, yielding CDelta170; (ii) introducing a six amino acid deletion in the N-terminal region of C, CDelta10-15; and (iii) combining these stable deletion mutations with the att CR84G/HNT553A mutation. The resulting rHPIV1 vaccine candidates were evaluated for attenuation in hamsters and AGMs and for immunogenicity and protective efficacy in AGMs. The CDelta10-15 mutation was attenuating in hamsters but not in AGMs, and likely will be of limited value for an HPIV1 vaccine. Conversely, the CR84G/HNT553A mutation set was attenuating in AGMs but not in hamsters. Thus, these two mutations demonstrated reciprocal host range phenotypes involving different regions of C. The CDelta170 mutation conferred a significant level of attenuation in hamsters and AGMs that closely resembled that of CF170S and will be of particular utility for vaccine development because it involves a deletion of six nucleotides rendering it highly refractory to reversion. The combination of the CR84G/HNT553A mutation set and the CDelta170 deletion mutation yielded a virus, rCR84G/Delta170 HNT553A, that exhibited a satisfactory level of attenuation in hamsters and AGMs and was immunogenic and highly protective against HPIV1 wt challenge. This virus will be evaluated clinically as a live intranasal HPIV1 vaccine, one that can be further attenuated as necessary by the introduction of additional stabilized att mutations previously developed in the L protein. PMID- 16364512 TI - A mutation inactivating the methyltransferase gene in avirulent Madrid E strain of Rickettsia prowazekii reverted to wild type in the virulent revertant strain Evir. AB - Rickettsia prowazekii Madrid E (E) strain is an effective vaccine, but can revert to virulent status when passaged in animals. The aim of this study is to identify the reverse mutation that may determine the virulence of R. prowazekii by comparing the genetic structures of E strain and its virulent revertant Evir strain. We determined that the gene (Rp028/Rp027) encoding the methyltransferase was mutated by frameshift in avirulent E strain but not in virulent revertant Evir strain and wild type virulent Breinl strain. We conclude that the mutation in the E strain gene reverts to wild type in the virulent revertant Evir strain. Whether the mutation plays an essential role in the attenuation of E strain needs to be further investigated. PMID- 16364513 TI - Development and evaluation of a Shigella flexneri 2a and S. sonnei bivalent invasin complex (Invaplex) vaccine. AB - Over 160 million cases of shigellosis occur annually worldwide, with the two most prevalent species being Shigella flexneri and S. sonnei. Protective immunity against Shigella infection is primarily directed at the lipopolysaccharide (LPS) of the homologous serotype, so it may be necessary to combine monovalent vaccines for multiple Shigella serotypes to construct a multivalent vaccine against predominant serotypes. Recently, we described a subcellular vaccine isolated from virulent S. flexneri, consisting of proteins (including the invasins IpaB and IpaC) and LPS, that protected mice and guinea pigs from homologous challenge. In the present study, a bivalent Invaplex vaccine consisting of S. flexneri 2a and S. sonnei Invaplex was used to intranasally immunize mice and guinea pigs to determine the bivalent vaccine's immunogenicity and protective capacity against challenge with either strain. Mice and guinea pigs immunized with the bivalent S. flexneri 2a/S. sonnei Invaplex vaccine produced serum IgA and IgG antibodies to S. flexneri LPS, S. sonnei LPS, the homologous Invaplex and the water extract antigens (invasins) as determined by ELISA. The immune responses in animals immunized with the bivalent vaccine were similar to responses in animals immunized with the monovalent Invaplex vaccines. Mice and guinea pigs immunized with the bivalent vaccine were protected from a lethal lung challenge (mice, P<0.001) or severe keratoconjunctivitis (guinea pigs, P< or = 0.002) after challenge with either S. flexneri 2a or S. sonnei. Animals immunized with monovalent Invaplex vaccines were protected (P<0.001) against the homologous agent at levels comparable to the bivalent vaccine. After challenge, immunized animals demonstrated boosts in antibody titers to LPS, water extract antigens and Invaplex. These studies indicate that the subcellular Invaplex vaccine will be readily adaptable to a multivalent vaccine approach for shigellosis. PMID- 16364514 TI - Analysis of costimulatory molecule expression on antigen-specific T and B cells during the induction of adjuvant-induced Th1 and Th2 type responses. AB - Previous studies show that the generation of maximal T cell responses requires B cell antigen presentation and the differential expression of costimulatory molecules by B cells may affect polarization of naive T cells to Th1 or Th2 phenotypes. We have therefore characterized the expression of activation and costimulatory molecules on antigen-specific T and B cells following immunisation with Alum or Alum/LPS to induce Th2 or Th1 responses in vivo. While antigen specific B cells show similar levels of activation with respect to MHCII upregulation following Th1 or Th2 induction, they differentially express costimulatory molecules. Although ICOS-B7RP-1 interactions were originally implicated in Th2 generation, surprisingly this receptor-ligand pair was only upregulated on antigen-specific T and B cells following Th1 induction. In conclusion, these studies indicate that during the generation of antigen-specific Th1 or Th2 responses, adjuvants induce differential costimulation in antigen specific B cells that may subsequently influence T cell polarization. PMID- 16364515 TI - Schwannomatosis: multiple schwannomas of the upper limb. AB - Schwannomas (also known as neurilemmomas) are neoplasms of the peripheral nerve sheaths. They are the most common benign tumour of peripheral nerves, yet they account for only 5% of all soft tissue tumours. Their incidence in the upper limb is variable, as is their clinical presentation. We present an unusual case of multiple schwannomas in the upper limb so that other surgeons can share our experience and suggest the correct diagnosis when confronted with a similar case. PMID- 16364516 TI - Effect of rotated head posture on dynamic vertebral artery elongation during simulated rear impact. AB - BACKGROUND: Elongation-induced vertebral artery injury has been hypothesized to occur during non-physiological coupled axial rotation and extension of head. No studies have quantified dynamic vertebral artery elongation during head-turned rear impacts. Therefore, we evaluated effect of rotated head posture vs. forward head posture at the time of impact on dynamic vertebral artery elongation during simulated rear impacts. METHODS: A whole cervical spine model with surrogate head and muscle force replication underwent either simulated head-turned (n = 6) or head-forward (n = 6) rear impacts of 3.5, 5, 6.5 and 8 g. Continuous dynamic vertebral artery elongation was recorded using custom transducer and compared to physiological values obtained during intact flexibility testing. FINDINGS: Average (SD) peak dynamic vertebral artery elongation of up to 30.5 (2.6) mm during head-turned rear-impact significantly exceeded (P < 0.05) the physiological beginning at 5 g. Highest peak elongation of 5.8 (2.1) mm during head-forward rear impact did not exceed physiological limit. Head-turned rear impact caused earlier occurrence of average peak vertebral artery elongation, 84.5 (4.2) ms vs. 161.0 (43.8) ms, and higher average peak vertebral artery elongation rate, 1336.7 (74.5) mm/s vs. 211.5 (97.4) mm/s, as compared to head forward rear impact. INTERPRETATION: Elongation-induced vertebral artery injury is more likely to occur in those with rotated head posture at the time of rear impact, as compared to head-forward. PMID- 16364517 TI - Intramuscular hemodynamics in bilateral erector spinae muscles in symmetrical and asymmetrical postures with and without loading. AB - BACKGROUND: Although attention has been paid to the relationship between the changes in blood circulation in erector spinae muscles and back pain, little is known about their hemodynamics in several various comparable postures with and without loading. Studies on hemodynamics of erector spinae muscles using near infrared spectroscopy have been performed on subjects and patients mainly in forward flexion positions. METHODS: Two near-infrared spectroscopes were used to measure oxygenated hemoglobin, deoxygenated hemoglobin, and total hemoglobin in bilateral erector spinae muscles at L2-3 in subjects in 9 postures, and holding no load, 10 kg or 20 kg in maximum flexed and lateral bending. Those three values in each posture and loading condition were expressed as a percentage of their corresponding values obtained in the standing upright position, and designated and statistically analyzed as %Oxy-Hb, %Deoxy-Hb and %Total-Hb, respectively. FINDINGS: %Total-Hb and %Oxy-Hb in maximum flexion were the most decreased. In maximum lateral bending, %Oxy-Hb only in the contralateral erector spinae muscles was decreased. When the load was 20 kg, the decreases in %Oxy-Hb were the largest in maximum flexion and lateral bendings. INTERPRETATION: Using two near-infrared spectroscopes allowed us to measure simultaneously the hemodynamics of bilateral muscles. They demonstrated different responses in each side. Asymmetrical posture and loading were accompanied by asymmetrical changes of the bilateral erector spinae muscles. Stretched muscle had less blood volume and oxygenation, both of which decreased with increasing load. These results showed that these postures and conditions might lead to fatigue of the ES muscles. PMID- 16364518 TI - The role of mesenchymal stem cells in haemopoiesis. AB - The ontogeny of haemopoiesis during fetal life and the differentiation of blood cells in adult life depend upon a fully competent microenvironment to provide appropriate signals via production of soluble factors and cell contact interactions. The cellular constituents of the microenvironment, also defined as the haemopoietic niche, largely derive from a common progenitor of mesenchymal origin. Mesenchymal stem cells (MSC), initially identified in adult bone marrow, have also been described in fetal haemopoietic tissues where they accompany the migration of haemopoietic development. Their precise identity remains ill-defined because of the lack of specific markers. Their ability to self-renew and differentiate into tissues of mesodermal origin (osteocytes, adipocytes, chondrocytes) and their lack of expression of haemopoietic molecules are currently the main criteria for isolation. In the bone marrow the most important elements of the niche appear to be osteoblasts, whilst a less defined population of fibroblasts regulates the maturation of immature T cells in the thymus. Recently, MSC have been shown to exert a profound immunosuppressive effect on polyclonal as well as antigen-specific T cell responses by inducing a state of division arrest anergy. Thus, the multipotent capacity of MSC, their role in supporting haemopoiesis, and their immunoregulatory activity make MSC particularly attractive for therapeutic exploitation. PMID- 16364519 TI - Human natural killer cell development and biology. AB - Natural killer cells are important innate immune effector cells with potentially broad applications in the treatment of human malignancy due to their ability to lyse neoplastic cells without the need for tumor-specific antigen recognition. Human NK cells can be divided into two functional subsets based on their surface expression of CD56; CD56(bright) immunoregulatory cells and CD56(dim) cytotoxic cells. In addition to functional differences, these NK cell subsets can be modulated differently by interleukin (IL)-2, which has permitted the development of lower dose, better tolerated IL-2 regimens for the in vivo expansion and activation of NK cells. The importance of early hematopoietic growth factors, such as c-kit ligand and flt-3 ligand, and their synergy with IL-15 in the development of human NK cells in the bone marrow has permitted the investigation of novel cytokine combinations for optimizing in vivo expansion of NK cell in the clinic. The importance of lymph nodes as a site for NK cell development has recently been elucidated. Furthermore, progress in the field of how NK cell recognize target cells via activating and inhibitory receptors, and how the balance of signals from these receptors can modulate NK cell activity has revolutionized our understanding of the selective killing of tumor cells by NK cells while sparing normal cells. In this review, we summarize current understanding of NK cell biology, and highlight how such knowledge may be translated to optimize the efficacy of using autologous or allogeneic NK cell for the immunotherapy of cancer. PMID- 16364520 TI - Metal concentrations in Kandalaksha Bay, White Sea (Russia) following the spring snowmelt. AB - Elevated concentrations of dissolved and particulate Cd, Cu, Pb and Zn have been determined in the waters of Kandalaksha Bay (White Sea, Russia), following the ice melt in the spring of 2000. Dissolved metal maxima in the surface waters were observed at some stations and concentrations generally decreased with depth. The suspended particulate matter (SPM) comprised a non-lithogenic fraction in the range 12-83%, and had elevated metal concentrations that showed no trend with depth or salinity and was compositionally distinct from the sediments. A log linear relationship existed between the concentrations of metals in sediments and in SPM and their respective Al concentrations, indicating a source of metal-rich particles, with low Al content, to the Bay. The results suggest that Kandalaksha Bay has been impacted by industrial activity on the Kola Peninsula and that restricted water exchange will hinder its recovery from metal contamination. PMID- 16364521 TI - Nitrogen balance and groundwater nitrate contamination: comparison among three intensive cropping systems on the North China Plain. AB - The annual nitrogen (N) budget and groundwater nitrate-N concentrations were studied in the field in three major intensive cropping systems in Shandong province, north China. In the greenhouse vegetable systems the annual N inputs from fertilizers, manures and irrigation water were 1358, 1881 and 402 kg N ha( 1) on average, representing 2.5, 37.5 and 83.8 times the corresponding values in wheat (Triticum aestivum L.)-maize (Zea mays L.) rotations and 2.1, 10.4 and 68.2 times the values in apple (Malus pumila Mill.) orchards. The N surplus values were 349, 3327 and 746 kg N ha(-1), with residual soil nitrate-N after harvest amounting to 221-275, 1173 and 613 kg N ha(-1) in the top 90 cm of the soil profile and 213-242, 1032 and 976 kg N ha(-1) at 90-180 cm depth in wheat-maize, greenhouse vegetable and orchard systems, respectively. Nitrate leaching was evident in all three cropping systems and the groundwater in shallow wells (<15 m depth) was heavily contaminated in the greenhouse vegetable production area, where total N inputs were much higher than crop requirements and the excessive fertilizer N inputs were only about 40% of total N inputs. PMID- 16364522 TI - Stressors and child and adolescent psychopathology: evidence of moderating and mediating effects. AB - This paper reviews studies that have tested for moderators or mediators of the relation between stressors and child and adolescent psychopathology. Many studies have tested for moderation, but results of research studying moderators have been inconclusive. There have been few theory-based studies and there have been few consistent findings. Far fewer studies have tested for mediation effects, but these studies have generally been theory-driven, have more often built upon one another in an incremental fashion, and have yielded consistent results. In particular, there is substantial evidence for the mediating role of family relationship in the relation between stressors and child and adolescent psychological symptoms. Future studies should integrate moderator and mediator research by testing for specific mediators in relation to particular moderating contexts, so that we can better understand the complex ways in which stressful life experiences affect the well-being of children and adolescents. PMID- 16364523 TI - Relapse to driving under the influence (DUI): a review. AB - Driving under the influence (DUI) is a major public health problem. In 2003, there were 17,401 alcohol-related crash fatalities. Although there has been a large decrease in the fatality rates over the past two decades, further progress has stalled in recent years. This plateau in the injury and death rates resulting from impaired driving has been attributed, in part, to the persistent or repeat DUI offender. Broadly defined, repeat offenders are those individuals who, following an initial DUI arrest, relapse to driving under the influence of alcohol and other drugs. In this paper, we first provide a brief overview of several models of DUI relapse. We then review the empirical literature on DUI relapse, the data describing characteristics of first-time and repeat DUI offenders, and, especially, studies that have evaluated the impact of legal sanctions and rehabilitation programs on subsequent DUI behavior. The data reveal that DUI offenders are a heterogeneous group, and that simple models relying on only one or two behavioral domains (e.g., driving characteristics, demographics) to explain DUI relapse are insufficient to account for the DUI behavior of offenders. To advance our understanding of DUI relapse, we argue for development and testing of multifactorial models focusing on the interplay of legal, social and psychological factors that describe and explain relapse among DUI offenders. By recognizing the heterogeneity within the offender population it will be easier for researchers and clinicians to identify subgroups that are at high-risk for relapse and which should be targeted by prevention and intervention programs. PMID- 16364524 TI - Relapse to alcohol and other drug use in treated adolescents: review and reconsideration of relapse as a change point in clinical course. AB - Relapse serves as an early warning sign of a failure to maintain desired behavioral change. Although rates of and time to relapse appear to be similar for treated adolescents and adults, this critical review of the literature on adolescent relapse begins with an overview of developmental differences between adolescent and adult substance users that need to be considered when studying relapse. A summary of rates of relapse in treated teens is followed by a review of results on the predictive validity of different relapse definitions. Next, characteristics of teens' initial relapse in terms of substances involved, situational context, and factors associated with relapse are described. Because relapse is ideally understood in the context of longer-term patterns of use, the review also discusses the extent to which early episodes of use mark clinically significant change points in post-treatment course, and how knowledge of longer term clinical course is essential to understanding the relapse process and mechanisms underlying the maintenance of behavioral change. PMID- 16364525 TI - Improved ELISA method for screening human antigen-specific IgE and its application for monitoring specific IgE for novel proteins in genetically modified foods. AB - For monitoring the occurrence of IgE antibody specific for novel proteins in genetically modified (GM) foods, ELISA is the most convenient method. The levels of IgE specific for recombinant proteins, phosphinothricin-N-acetyltransferase (PAT), CP4-EPSPS, and Cry9C were determined by ELISA using the sera from patients allergic to known allergens. Ovalbumin (OVA) and OVA-positive patient sera were used as positive control. In the ELISA, 20-fold-diluted sera tested were mostly negative for the specific IgE. However, the PAT-specific, but not CP4-EPSPS- or Cry9C-specific IgE in some patients was apparently higher than that of the healthy volunteers. To clarify the binding specificity of the antibody, we pre incubated the sera with soluble PAT, but the inhibition was marginal, suggesting that the binding was non-specific. Therefore, we used 1M NaCl as a washing buffer to remove IgE non-specifically bound to the coated PAT. This washing step efficiently decreased non-specific binding. In contrast, OVA-specific IgE binding to OVA-coated plate was not affected by the washing. Finally, in this pilot study significant levels of IgE antibodies specific for the three proteins were not detected in the sera of Japanese food-allergy patients. PMID- 16364526 TI - Does socio-economic advantage lead to a longer, healthier old age? AB - The effect of socio-economic disadvantage on mortality is well documented and differences exist even at older ages. However, whether this translates into differences in the quality of life lived at older ages is less well studied, and in particular in the proportion of remaining life spent without ill health (healthy life expectancy), a key UK Government target. Although there have been studies exploring socio-economic differences in disability-free life expectancy (DFLE) worldwide, these have tended to focus on a single measure of socio economic advantage, for example, education, race, social class or income, with the majority based on cross-sectional data from younger populations. In this prospective study we examine differences in DFLE and total life expectancy (TLE) at older ages using a range of measures of socio-economic advantage. We use a longitudinal study of 1480 participants aged 75 years or over in 1988 registered with a UK primary care practice, who were followed up until 2003 with measurements at up to seven time points. Disability was defined as difficulty with any one of five activities of daily living. The largest differences in DFLE for both men and women were found for housing tenure. Women aged 75 years living in owned or mortgaged property could expect to live 1 year extra without disability compared with those living in rented accommodation, while for men the difference was almost 1.5 years. The effect of socio-economic advantage on disability-free and total life expectancies appeared to be larger for men than women. In women, socio-economic advantage had more effect on DFLE than total life expectancy for all indicators considered, thus the socio-economically advantaged experienced a compression of disability. PMID- 16364527 TI - Could the peer group explain school differences in pupil smoking rates? An exploratory study. AB - Schools differ in the proportion of their pupils who smoke. Such differences transcend pupil intake characteristics and relate to the internal life of the school. Although adolescents' smoking behaviour has been associated with that of their peers, little consideration has been given to whether peer structures and processes contribute to school differences in pupil smoking rates. In two relatively deprived Scottish schools, one with a higher and one with a lower rate of pupil smoking, 13 and 15 year-olds were surveyed. Sociometric data and information on pupils' smoking behaviour and views were gathered. Twenty-five single-sex discussion groups were then held with a sub-sample of the 13 year-olds in order to explore in detail their views on smoking, smokers and fellow pupils. Findings showed that in the higher smoking school, pupils were more often in groups, smokers were identified as popular, and attitudes (especially among non smoking females) were more pro-smoking. In the lower smoking school, by contrast, there were more isolates and dyads, there were no popular smokers and attitudes (especially among non-smoking females) were much less pro-smoking. Thus, evidence suggests peer group structures and related influences could be one explanation for school differences in smoking, and that the popularity of smokers together with the views of non-smoking females may be particularly important in creating such differences. PMID- 16364528 TI - Legal and ethico-legal issues in e-healthcare research projects in the UK. AB - This report first gives an overview of e-healthcare research activity in the UK. It then summarises available information concerning legal hurdles and issues faced by these projects, provides an overview of work being carried out to investigate these hurdles, and considers some of the proposals put forward to overcome them. It is an initial scoping review (using both literary and web-based resources)-rather than a systematic or analytical review-of e-health which addresses specific strands relating to legal and ethico-legal issues and identifies activity in these areas. It is hoped that this preliminary scoping exercise will stimulate further research and analysis. PMID- 16364529 TI - Latencies of stimulus-driven eye movements are shorter in dyslexic subjects. AB - Eye movements latencies toward peripherally presented stimuli were measured in 10 year-old dyslexic and control children. Dyslexic subjects, previously found to be oversensitive to stimulation of the magnocellular channel, showed reduced latencies as compared to normally reading controls. An attention shifting task was also used which showed no group differences in latencies of eye movements. The data are discussed in terms of the hypothesis of magnocellular system alteration and attention dysfunction in dyslexia. Additionally, sex differences in eye movement latencies were found and are discussed. PMID- 16364530 TI - Exploration for unknown substances in rapeseed oil that shorten survival time of stroke-prone spontaneously hypertensive rats. Effects of super critical gas extraction fractions. AB - To identify the causative substances for the shortening of survival time by rapeseed (Canola) oil in stroke-prone spontaneously hypertensive rats (SHRSP), SHRSP were fed on a standard chow supplemented with 10 w/w% soybean oil (control), rapeseed oil, one of the fractions of rapeseed oil obtained by super critical gas extraction (SCE) under a pressure of 180-bar or 350-bar, at 40 degrees C, or the residue from the extraction (with 0.5% NaCl in drinking water). In another series of experiment, SHRSP were fed for 8 weeks on the above mentioned diets without salt loading and autopsied. Fatty acid compositions in these diets were similar, except in the soybean oil diet, and phytosterol contents were: (diet containing) 180-bar fraction>residue>rapeseed oil>350-bar fraction>soybean oil. Survival times in the rapeseed oil, 350-bar fraction and residue groups were shorter than, whereas that in the 180-bar fraction was similar to in the soybean oil group. In the 8-week feeding experiment, chronic nephropathy was found frequently in the groups other than the soybean oil group. The heart weights were higher in the rapeseed oil and residue groups. Cerebral necrosis was found in the residue group. Taken together, the followings are concluded, (1) Neither the fatty acid composition, nor the amount of phytosterols in the diets appeared to be decisive in the shortening of life. (2) SCE appeared to produce a safe (180-bar) fraction, though it failed to separate clearly the causative substances into specific fractions. (3) The factors that facilitate the genetic disease of SHRSP appear to exist in rapeseed oil. However, they might not be identical to those responsible for the life-shortening, since there were no findings common across the rapeseed oil, 350-bar and residue groups, which showed similar life-shortening. PMID- 16364531 TI - Effects of ethanol and diphenyl diselenide exposure on the activity of delta aminolevulinate dehydratase from mouse liver and brain. AB - Ethanol toxicity is affected by both environmental and inherited features. Since oxidative stress is an important molecular mechanism for ethanol-induced cellular damage, the concomitant exposure to ethanol and pro-oxidative or antioxidant compounds can alter its toxicity. Here, we investigate the effects of exposure to ethanol and/or diphenyl diselenide, an organochalcogen with antioxidant properties, on parameters related to oxidative stress (thiobarbituric acid reactive species-TBARS-and delta-aminolevulinate dehydratase-delta-ALA-D activity) in mouse liver and brain. In addition, the in vitro effects of ethanol and acetaldehyde on the activity of delta-ALA-D from human erythrocytes were also investigated. Both ethanol and diphenyl diselenide decreased hepatic delta-ALA-D activity and DL-dithiothreitol (DTT) reactivated this enzyme only after ethanol induced inhibition. Moreover, ethanol increased liver TBARS levels, independently of the presence of diphenyl diselenide treatment. Brain delta-ALA-D activity and TBARS levels were not changed by ethanol or diphenyl diselenide exposure. Under in vitro conditions, acetaldehyde was a more potent inhibitor of delta-ALA-D from human erythrocytes when compared to ethanol, demonstrating a dose-dependent effect. This study indicates that (1) hepatic delta-ALA-D is a molecular target for the damaging effect of ethanol under in vivo conditions; (2) diphenyl diselenide and ethanol seem to inhibit delta-ALA-D by different mechanisms; (3) acetaldehyde, a metabolite of ethanol, is probably the main molecule responsible for the inhibitory effects of the parent compound on delta-ALA-D. PMID- 16364532 TI - Safety studies on epigallocatechin gallate (EGCG) preparations. Part 1: genotoxicity. AB - Public interest in green tea has grown recently due to the potential health benefits from its consumption. Epigallocatechin gallate (EGCG), a principal polyphenolic component of green tea, is considered key to these healthful qualities. Although numerous studies have evaluated the anti-cancer effects of green tea and EGCG, few have examined the safety of EGCG consumption. The genotoxic potential of a concentrated EGCG preparation was tested in Salmonella and L5178Y tk+/- mouse lymphoma cell assays to further define the safety of Teavigo, a high-concentration EGCG extract of Camellia sinensis leaves produced by the same novel method. No mutagenic activity was detected in the bacterial system; however, a clastogenic 'trend' from the formation of hydrogen peroxide was noted in the murine cells. The oral administration of 500, 1000, or 2000 mg EGCG/kg to mice did not induce micronuclei formation in bone marrow cells. Similarly, administering 400, 800, or 1200 mg EGCG/kg/day in their diet for 10 days did not induce bone marrow cell micronuclei and produced plasma EGCG concentrations comparable to those reported in human studies. The intravenous injection of 10, 25 and 50 mg EGCG/kg/day to rats resulted in much higher plasma concentrations and demonstrated an absence of genotoxic effects. From these studies, it is concluded that Teavigo (EGCG) is not genotoxic. PMID- 16364533 TI - Genotoxic potential of by-products in drinking water in relation to water disinfection: survey of pre-ozonated and post-chlorinated drinking water by Ames test. AB - Mutagenic potential of drinking water samples derived from ranneywells was studied. 100-100 l of untreated (rough) and ozone-treated as well as chlorinated disinfected water were dropped on and adsorbed by macroreticular resin columns (Serdolit PAD-III and Amberlite XAD-2). The adsorbed material was desorbed by methanol and dichloromethane. After elimination of the solvents by vacuum distillation the adsorbed material was dissolved in dimethylsulfoxide. The mutagenic activity was tested in the Ames-Salmonella/rat liver microsome system. The tester strains were TA-98 and TA-100. The material adsorbed to Serdolit PAD III from rough and also disinfected water did not induce mutagenicity in case of the TA-98 tester strain, irrespective of activation by liver microsomes. However, the material adsorbed to Amberlite XAD-2 exerted mutagenic effect on the TA-98 tester strain, with and without liver microsome activation, both in case of rough and disinfected water. The TA-100 tester strain showed mutation after, but not without activation, when treated with the material adsorbed by either Serdolit PAD-III or Amberlite XAD-2, in case of rough water. Material derived from disinfected water and adsorbed to Serdolit PAD-III, caused mutation of the TA tester strain also only after activation. The material derived from disinfected water and adsorbed to Amberlite XAD-2 proved to be mutagenic to the TA-100 tester strain both without and after activation. Mutagenic activity was exerted by the amount of concentrates derived from 0.28 to 0.83 l of rough and 0.83-2.5l of disinfected water. The mutagenic activity of drinking water raises the possibility of carcinogenic effect, too. Search for alternative methods of water disinfection may be recommended. PMID- 16364534 TI - Conformation preserved in a weak-to-strong or strong-to-weak [PSI+] conversion during transmission to Sup35 prion variants. AB - The cytoplasmic [PSI(+)] element of budding yeast represents the prion conformation of translation release factor Sup35. Much interest lies in understanding how prions are able to generate variation in isogenic strains. Recent observations suggest that a single prion domain, PrD, is able to adopt several conformations that account for prion strains. We report novel PrD variants of Sup35 that convert weak [PSI(+)] to strong [PSI(+)], and vice versa, upon transmission from wild-type Sup35. During the transmission from wild-type Sup35 to variant Sup35s, no conformational changes were detected by proteolytic fingerprinting and the original [PSI(+)] strain was remembered upon return to wild-type Sup35. These findings suggest that during transmission to variant Sup35s, the [PSI(+)] phenotype is variable while the original conformation is remembered. A mechanism of "conformational memory" to remember specific [PSI(+)] conformations during transmission is proposed. PMID- 16364535 TI - Suitability of the vegetation types in Mexico's Tamaulipas state for the siting of hazardous waste treatment plants. AB - A land suitability study was carried out by applying a multiple-criteria technique to 12 different vegetation types in Mexico's Tamaulipas state to help select potentially suitable sites for hazardous waste treatment plants. Species richness, spatial distribution, and uniqueness were selected as the criteria for estimating a vegetation type's suitability. Using the analytical hierarchy process, we ranked and mapped vegetation types, then compared the results with rankings of the same vegetation types based only on their number of endemic species. The suitabilities of the various vegetation types were ordered in more or less the same way by both methods, except in two cases for which the results were very different. The method proved to be a useful tool despite the availability of only partial (mostly qualitative) information; under such circumstances, expert experience can be incorporated in the evaluation process to a limited degree. The technique described in this paper has a high potential to aid decisions when many opinions and options must be considered simultaneously. PMID- 16364536 TI - Laparoscopic sentinel lymph node dissection--a novel technique for the staging of prostate cancer. AB - OBJECTIVES: Pelvic lymph node metastases indicate a poor prognosis for prostate cancer patients. The aim of this study was to evaluate the suitability of laparoscopic radioisotope guided sentinel lymph node (SLN) dissection in staging of prostate carcinoma. METHODS: 28 patients with prostate cancer and intermediate or high risk for lymph node metastases considered for external beam radiotherapy underwent laparoscopic pelvic lymphadenectomy at our institution. For visualization of individual SLN distribution, an image fusion system consisting of a gamma-camera with integrated X-ray tube was used. During laparoscopic lymphadenectomy, SLN were identified using a laparoscopic gamma probe. RESULTS: Preoperative imaging and laparoscopic gamma probe allowed an excellent delineation of SLN. 57% (preoperative imaging) as well as 48% (intraoperative measurements) of SLN were found outside the obturator fossa. All SLN were removed successfully without intra- or postoperative complications. Despite extended lymphadenectomy, no significant lymphocele appeared. 10 lymph node metastases were found in 7 out of the 31 patients (23%) with 3 of the 10 metastases lying outside the obturator fossa representing the standard lymphadenectomy area. CONCLUSIONS: The present data demonstrate that laparoscopic SLN dissection is an excellent minimally invasive and technically feasible tool for staging of intermediate and high risk prostate cancer. PMID- 16364537 TI - Impact of race, age, income, and residence on prostate cancer knowledge, screening behavior, and health maintenance in siblings of patients with prostate cancer. AB - OBJECTIVE: This study evaluates self-reported changes in knowledge of prostate cancer (CaP), CaP screening behavior and other health-related activities in men whose brothers have been diagnosed with prostate cancer and to characterize those demographic subgroups of siblings at particular risk of failure to change their behavior. METHODS: 112 men were surveyed to self-assess their knowledge of CaP both before and after their brother's diagnosis. In addition, siblings were questioned with regard to other health-related behaviors before and after their brother's diagnosis. Demographic characteristics including age, race, income, and area of residence were also reported. Results were analyzed to determine how a brother's diagnosis with CaP affects changes in prostate cancer knowledge, screening behavior and other health related activities in these high-risk patients. RESULTS: The impact of a brother's diagnosis of CaP affects only 40% of siblings with regard to CaP knowledge. This knowledge appears to translate into increased screening behavior and other-health-related activity in these men. Unfortunately, more than half of men have no change in CaP knowledge and correspondingly no change in screening behavior. Siblings who are older, earn less, and live in rural areas have a lower baseline knowledge of CaP and are less likely to improve their self-assessed knowledge. On regression analysis, correlates of improvement in CaP knowledge included (1) those discussing with their primary physician (OR=6.6), (2) Caucasians (OR=2.7) and (3) non-smokers (OR=3.1). Improvements in prostate cancer knowledge were found to be predictive of subsequent participation in CaP screening and annual physical exams. CONCLUSIONS: Increased attention should be paid to siblings of CaP patients in efforts to improve their knowledge and thereby participation in screening as these patients are at an increased risk of development of this disease. Efforts should be made to educate siblings who are older, of lower income, African American, and rural residents with regard to CaP, as these subgroups appear less likely to change their knowledge and screening behavior despite their sibling's diagnosis. PMID- 16364538 TI - Cysteine 390 mutation of the TSH receptor modulates its ectodomain as an inverse agonist on the serpentine domain with decrease in basal constitutive activity. AB - Mutations of individual cysteine residues at codon 301, 390, 398 and 408 of the thyrotropin receptor (TSHr) to serine resulted in cell surface expression of only C301S and C390S mutants. C390S mutation was a silencing mutation with decreased basal constitutive activity. Although the C301S and C390S mutants did not show any significant TSH binding, they generated cyclic AMP upon TSH stimulation. These mutants were also able to interact with stimulating and blocking anti-TSHr antibodies. In fact, C390S receptor is a more sensitive tool for blocking antibody detection than wild type receptor. Introduction of C390S to activating mutations in the ectodomain (S281N), exloop (I486F) and transmembrane (D633H) segments could not mute/nullify receptor activation. These data indicate that the C390S ectodomain behaves as a more effective inverse agonist on the noisy transmembrane segment and suggest that the basal and activated states of the receptor operate through two independent pathways. PMID- 16364539 TI - Psychiatric manifestations of multiple sclerosis and acute disseminated encephalomyelitis. AB - It is unusual for acute disseminated encephalomyelitis and multiple sclerosis to present as purely psychiatric disorders. We report five patients with such demyelinating diseases and symptoms of psychosis, depression or anxiety. The importance of excluding demyelination as the basis for these psychiatric disturbances is emphasized, especially in the presence of unexplained neurologic findings. The possible relationship between psychiatric symptoms and demyelinating disorders is explored. PMID- 16364540 TI - The impact of the international subarachnoid aneurysm trial (ISAT) on the management of aneurysmal subarachnoid haemorrhage in a neurosurgical unit in the UK. AB - OBJECTIVE: To review the changes in the management of aneurysmal subarachnoid haemorrhage (SAH) in a single neurosurgical unit in the UK, following the publication of the international subarachnoid aneurysm trial (ISAT). METHODS: The presentation, investigations, treatments and outcome data of all patients admitted with SAH to the neurosurgical unit between February 2001 and May 2003 were prospectively recorded in a database. The total period studied was split in to three blocks, around the time of publication of the ISAT in October 2002 (period 1=February-December, 2001; period 2=January-September, 2002 and period 3=October 2002 to May 2003). RESULTS: Of the 177 patients admitted with presumed SAH, 130 patients with evidence of an aneurysm on angiograms were included in the study. The mean age was 53+/-1 years, 92 (71%) patients were WFNS grade 1 or 2 and 77 (60%) were Fischer grade 2 or 3. These parameters were unchanged over the study period. Overall, 60 patients (46%) underwent a craniotomy for clipping or wrapping of aneurysm, 60 (46%) underwent endovascular embolisation of the aneurysm and 10 patients (8%) were managed conservatively. Over the study periods 1-3, the proportion of patients undergoing open surgery decreased (from 51 to 31%) while endovascular treatment of aneurysms increased (35-68%; p<0.01). Over the same time points there was a non-significant trend towards better Glasgow outcome scores at 6 months follow-up. The management mortality for all WFNS grades of patients with SAH was eight deaths (14%). The mortality in the surgical group was 3 patients (5%) and there were no deaths in the endovascular group. Over the study periods 1-3, there was a decrease in the mean total duration of hospital stay (from 23.6 to 15.5 days; p<0.05) in WFNS grade 1 and 2 patients and this was related to a shorter duration of hospital stay in the endovascular than surgical group of patients (p<0.05). The mean delay in obtaining an angiogram increased over the study periods 1-3 (1.1-2.3 days; p<0.05). CONCLUSIONS: This observational study highlights the changing pattern of management of SAH and the potential difficulties that could be encountered. The proportion of patients undergoing endovascular treatment of aneurysms has increased following the publication of the ISAT study. The associated increase in the delay in obtaining an angiogram may reflect the increased workload encountered by the neuroradiologists. PMID- 16364541 TI - Early cellular events in multiple sclerosis. Intimations of an extrinsic myelinolytic antigen. AB - In a previous immunohistological study of tissues from unusually early cases of MS cluster analysis revealed a progression of demyelination through five distinct stages [Gay F, Drye T, Dick G, et al. The application of multifactorial cluster analysis in the staging of plaques in early multiple sclerosis. Identification and characterization of the primary demyelinating lesion. Brain 1997;120:1461 83]. Tissues from six of the earliest cases in this series contained regions of normal appearing white and grey matter in which well developed inflammatory events, concentrated in perivascular spaces, were found to extend locally into the perivascular parenchyma to envelop ostensibly intact myelin sheaths. The beginnings of myelin sheath lysis and phagocytosis were subsequently detected within these lesions and similar foci were found in subpial and in subependymal tissues. They were characterised by a spreading HLA Class II antigen expression on microglia, and by the presence of co-locating C3 complement-IgG complexes on capillary basement membranes, on microglial cell membranes and within the cytoplasm of large bodied activated astrocytes. Parenchymal lesions contained significantly few CD4+ T cells and showed no evidence of capillary leakage of plasma proteins. Despite the presence of complexed immunoglobulin and complement, opsonization of the myelin sheath could not be demonstrated. These observations point to the presence in early MS of a diffusing, complement-fixing, myelinolytic antigen, processed mainly within the Virchow-Robin spaces and distributed in the cerebrospinal and extracellular fluid compartments of the central nervous system. PMID- 16364542 TI - Acute limbic encephalitis: a new entity? AB - Clinical cases similar to herpes simplex virus (HSV) encephalitis have accumulated in Japan. Detailed examinations have failed to demonstrate HSV infection. Recently, these cases have been named "non-herpetic acute limbic encephalitis". Only a single autopsy case was so far reported in an abstract form, because many cases showed a good prognosis. The case presented here was that following fever, a 59-year-old woman developed disturbance of consciousness and uncontrollable generalized seizures. Brain MRI revealed abnormal signals in the bilateral medial temporal lobe and along the lateral part of the putamen. Autoantibody against the NMDA glutamate receptor (GluR) IgM-epsilon2 was detected in the serum, and the GluR IgG-delta2 antibody was positive in cerebrospinal fluid. She died 12 days after onset. An autopsy examination revealed scattered foci consisting of neuronal loss, neuronophagia and some perivascular lymphocytic infiltration in the hippocampus and amygdala, but no haemorrhagic necrosis in the brain. HSV-1, -2 and human herpes virus-6 were negative immunohistochemically. We believe that our autopsy case may contribute to understanding the neuropathological background of non-herpetic acute limbic encephalitis. PMID- 16364543 TI - Expression profile of heat shock protein 108 during retinal development in the chick. AB - In the developing chick retina, heat shock protein 108 (HSP108), which exhibits transferrin binding activity, has been demonstrated at the mRNA level, while transferrin shows two expression peaks. Here, we investigated the expression profile of HSP108 in the developing chick retina at the protein level. The localization of HSP108 in embryonic days 15 (E15), E18, and postnatal day 2 (P2) chick retina was examined immunohistochemically using monoclonal antibody 9G10 specific for chick HSP108, while the expression levels of HSP108 in developing chick retina from E12 to P2 and adult were measured by Western blot analysis. HSP108 was expressed in the ganglion cell layer, inner nuclear layer, outer plexiform layer, outer nuclear layer, inner segments of photoreceptors and retinal pigment epithelium. Two peaks of HSP108 expression were found at around E13 and E18, respectively. Since the two HSP108 peaks appeared to be correlated with the transferrin expression peaks during retinal development, HSP108 may be associated with iron metabolism during the development of the retina. PMID- 16364544 TI - Fractality of dendritic arborization of spinal cord neurons. AB - Skeletonized images of Golgi impregnated neurons from the human, monkey, cat and rat dorsal horns were subjected to fractal analysis. These neurons have sparse branching of dendrite arbors. It is noticed that, in certain neuronal samples, some authors report that scaling range of experimentally declared fractals is extremely limited and spanned approximately between 0.5 and 2.0 decades. In order to retain our hypothesis that neurons with dendrites of uncomplicated shapes can be considered fractal over three decades of scale, we conducted four procedures: (i) we used the box-counting method, (ii) we scaled the box sizes as a power of 2, (iii) we chose the coefficient of correlation, measuring the "goodness of fit" of experimental data points to regression straight line, to be equal to or larger than 0.995, and (iv) we pointed out that all the neurons analyzed have a single fractal dimension measuring a global fractality showing no linear regions. As a control, we used some cerebellar Purkinje cells whose dendrite trees show much more complex structure and profuseness of branching. Since, generally, the neuronal structure is among the most complex of all cellular morphologies, we believe that supporting this hypothesis we advance the neuroscience and fractal theory. PMID- 16364545 TI - Effects of fluoxetine on cellular immune response in stressed mice. AB - We studied the effects of fluoxetine, a non-tricyclic antidepressant drug that selectively inhibits re-uptake of serotonin by presinaptic neurons in the brain, on cellular immune responses in mice exposed to a chronic auditory stressor. The natural killer (NK) cell activity was reduced after 4, 8, 12, 16 and 20 days of stress exposure with a partial recovery on days 16 and 20. Daily treatment with fluoxetine partially reversed these adverse effects of stress in a dose-dependent manner. Significant differences appeared when fluoxetine was administered at 2 mg/kg and maximum effect was reached at doses of 5 mg/kg. The capacity of T cells to generate cytotoxic T-lymphocytes (CTL) in mixed lymphocyte cultures and in vivo was reduced after 4 days of stress application and this effect was partially reduced when mice were injected with 5 mg/kg of fluoxetine. Nevertheless, in our experiments, fluoxetine did not significantly affect the cellular immunity in unstressed mice. In conclusion, fluoxetine seems to partially recover the adverse effects of chronic stress on cellular immune response. PMID- 16364546 TI - Bcl-2 immunoreactivity in human cutaneous Meissner and Pacinian corpuscles. AB - The occurrence and distribution of Bcl-2, a protein involved in the death-life cell pathways, was investigated in the peripheral sensory nervous system of healthy adult humans, including lumbar dorsal root ganglia, nerve trunks and glabrous skin (to analyze sensory corpuscles) using Western blot and immunohistochemistry. The antibody used labelled a protein of 26 kDa of estimated molecular weight corresponding with Bcl-2. Immunohistochemistry showed that only a neuronal population in dorsal root ganglia, some axons in peripheral nerves and the axon supplying Meissner and Pacinian corpuscles contained Bcl-2, whereas peripheral glial cells (i.e. satellite glial cells, Schwann cell, and lamellar cells of sensory corpuscles) did not. These results suggest that in normal conditions, Bcl-2 is only present in some neuronal, but not glial, elements of the sensory peripheral nervous system. The functional significance, if any, of these results remains to be determined. PMID- 16364547 TI - Role of PEPT2 in glycylsarcosine transport in astrocyte and glioma cultures. AB - The aims of the current study were (1) to quantify the role of PEPT2 in the uptake of glycylsarcosine (GlySar) in cultured neonatal astrocytes and (2) to examine GlySar transport and PEPT2 expression in two glioma cell lines. The uptake of [(14)C]GlySar was measured in astrocytes cultured from neonatal mouse (PEPT2(+/+) and PEPT2(-/-)) and rat, as well as rat C6 and F98 glioma cells. PEPT2 expression was examined by reverse transcription-polymerase chain reaction (RT-PCR). Neonatal astrocytes from PEPT2(-/-) mice had a 94% reduction in [(14)C]GlySar uptake compared to wild type mice and there was no saturable transport. In PEPT2(+/+) mice, [(14)C]GlySar uptake was saturable (V(max) 58 +/- 12 pmol/mg/min, K(m) 107 +/- 46 microM, K(d) 0.043 +/- 0.004 microl/mg/min). In neonatal rat astrocytes, kinetic analysis also suggested that [(14)C]GlySar uptake was via a single transporter. The inhibitor profile and pH dependence of that transport process was consistent with PEPT2. In C6 and F98 glioma cells, [(14)C]GlySar uptake was markedly reduced ( approximately 96-98%) compared to that in neonatal astrocytes and this was reflected by an absence of PEPT2 mRNA expression. These results indicate that PEPT2 is the sole transporter involved in the uptake of GlySar into neonatal cultured astrocytes. However, PEPT2 mRNA appears to be absent from two glioma cell lines. PMID- 16364549 TI - Mechanisms of placebo analgesia: rACC recruitment of a subcortical antinociceptive network. AB - Placebo analgesia is one of the most striking examples of the cognitive modulation of pain perception and the underlying mechanisms are finally beginning to be understood. According to pharmacological studies, the endogenous opioid system is essential for placebo analgesia. Recent functional imaging data provides evidence that the rostral anterior cingulate cortex (rACC) represents a crucial cortical area for this type of endogenous pain control. We therefore hypothesized that placebo analgesia recruits other brain areas outside the rACC and that interactions of the rACC with these brain areas mediate opioid-dependent endogenous antinociception as part of a top-down mechanism. Nineteen healthy subjects received and rated painful laser stimuli to the dorsum of both hands, one of them treated with a fake analgesic cream (placebo). Painful stimulation was preceded by an auditory cue, indicating the side of the next laser stimulation. BOLD-responses to the painful laser-stimulation during the placebo and no-placebo condition were assessed using event-related fMRI. After having confirmed placebo related activity in the rACC, a connectivity analysis identified placebo dependent contributions of rACC activity with bilateral amygdalae and the periaqueductal gray (PAG). This finding supports the view that placebo analgesia depends on the enhanced functional connectivity of the rACC with subcortical brain structures that are crucial for conditioned learning and descending inhibition of nociception. PMID- 16364548 TI - Neural correlates of individual differences in pain-related fear and anxiety. AB - Although individual differences in fear and anxiety modulate the pain response and may even cause more suffering than the initiating physical stimulus, little is known about the neural systems mediating this relationship. The present study provided the first examination of the neural correlates of individual differences in the tendency to (1) feel anxious about the potentially negative implications of physical sensations, as measured by the anxiety sensitivity index (ASI), and (2) fear various types of physical pain, as indexed by the fear of pain questionnaire (FPQ). In separate sessions, participants completed these questionnaires and experienced alternating blocks of noxious thermal stimulation (45-50 degrees C) and neutral thermal stimulation (38 degrees C) during the collection of whole-brain fMRI data. Regression analyses demonstrated that during the experience of pain, ASI scores predicted activation of a medial prefrontal region associated with self-focused attention, whereas FPQ scores predicted activation of a ventral lateral frontal region associated with response regulation and anterior and posterior cingulate regions associated with monitoring and evaluation of affective responses. These functional relationships cannot be wholly explained by generalized anxiety (indexed by STAI-T scores), which did not significantly correlate with activation of any regions. The present findings may help clarify both the impact of individual differences in emotion on the neural correlates of pain, and the roles in anxiety, fear, and pain processing played by medial and orbitofrontal systems. PMID- 16364551 TI - An epidemiological study of gastrointestinal parasites of dogs from Southern Greater Buenos Aires (Argentina): age, gender, breed, mixed infections, and seasonal and spatial patterns. AB - A total of 2193 fecal samples from owned dogs were collected during the 2003-2004 period in Southern Greater Buenos Aires, and were evaluated for the presence of intestinal parasites by a flotation-centrifugation method. The overall prevalence was 52.4%, and the 11 species found were: Ancylostoma caninum (13%), Isospora ohioensis complex (12%), Toxocara canis (11%), Trichuris vulpis (10%), Sarcocystis sp. (10%), Giardia duodenalis (9%), Isospora canis (3%), Hammondia Neospora complex (3%), Dipilydium caninum (18 cases), Cryptosporidium sp. (5 cases), and Toxascaris leonina (1 case). There was no significant difference in the overall prevalence between genders (female = 50.4%, male = 54.6%), and breeds (pure = 52.3%, mixed = 53%), but prevalence in puppies (<1 year) was higher than in adult dogs (62.7% versus 40.8%, respectively). Only the prevalence of A. caninum differed between genders, with higher values for males. The prevalences of six of the parasite species showed a decreasing trend with increasing host age, and an inverse pattern was found for two other species. The prevalences of three protozoa were significantly higher in pure-breed dogs, and those of two nematodes were significantly higher in mixed-breed dogs. The prevalences of T. canis, A. caninum, and T. vulpis were spatially heterogeneous with a clear Southwest-Northeast gradient. Only prevalences of Sarcocystis sp. and G. duodenalis showed seasonal variation. The frequency distribution of the number of species per fecal sample did not differ from a random distribution. Results obtained throughout the world were discussed. PMID- 16364553 TI - The utility of the functional independence measure (FIM) in discharge planning for burn patients. AB - Determining burn patients' need for inpatient rehabilitation at discharge is difficult and an objective clinical indicator might aid in this decision. The functional independence measure (FIM) is a validated outcome measure that predicts the need for rehabilitation services. This study evaluated the utility of the FIM score for discharge planning in burn patients. A retrospective chart review and FIM score determination was performed on all major burn patients discharged from a regional adult burn centre between July 1, 1999 and June 30, 2000. From 164 adult burn patients discharged, 37 met the American Burn Association criteria for major burns. One patient had insufficient data. Therefore, 36 patients were studied (mean age 47.3 +/- 17.4 years, and mean body area burned 27.4 +/- 12.9%). All 17 patients with FIM scores greater than 110 were discharged home, and patients with FIM score of 110 or lower were discharged to another institution (rehabilitation hospital n = 14, other acute care hospital n = 4, or a nursing home n = 1) p < 0.0001. A discharge FIM score of 110 or lower was strongly associated with the need for inpatient rehabilitation, while a FIM score greater than 110 indicates the patient is independent enough to manage at home. Further prospective studies will be necessary to validate these findings. PMID- 16364554 TI - Identification of lumbar spinal neurons controlling simultaneously the prostate and the bulbospongiosus muscles in the rat. AB - Lumbar spinothalamic neurons in the lamina X of the L3-L4 spinal cord segment have been proposed to constitute the spinal ejaculation generator in male rats. Lumbar spinothalamic cells are immunoreactive for galanin and neurokinin-1 receptors. We previously showed that after injection of pseudorabies virus either in the bulbospongiosus muscle or in the prostate, retrogradely labeled cells in the L3-L4 segment also displayed galanin or neurokinin-1 receptor immunoreactivities, demonstrating a direct link between lumbar spinothalamic cells and two anatomical structures involved in the two phases of ejaculation i.e. the emission and the expulsion phases. In order to provide with a more precise anatomical support for the role of lumbar spinothalamic cells in controlling ejaculation, we injected simultaneously in male adult rats two strains of recombinant pseudorabies virus, expressing either beta-galactosidase (PRV-BaBlu) or green fluorescent protein (PRV-152) in the prostate and in the bulbospongiosus muscle, respectively. After 5 days, we performed multiple immunofluorescence experiments to detect PRV-BaBlu, PRV-152 and galanin or neurokinin-1 receptors in transverse sections of the L1-S1 segment. Double- and triple-labeled cells were counted using confocal laser scanning microscope. Double-labeled neurons with the two strains of pseudorabies virus were mainly found at the L3-L4 segment lateral to the central canal in lamina X and represented about 60% of the total number of pseudorabies virus-labeled neurons. All the double pseudorabies virus-labeled neurons also expressed lumbar spinothalamic and most of them neurokinin-1 receptor, identifying them as lumbar spinothalamic neurons. The convergence of retrograde labeling from prostate and bulbospongiosus muscle on the same lumbar spinothalamic cells strongly reinforce their role in the spinal control and coordination of the emission and expulsion of sperm. PMID- 16364555 TI - Alterations in cortisol negative feedback inhibition as examined using the ACTH response to cortisol administration in PTSD. AB - OBJECTIVE: Studies using the dexamethasone suppression test (DST) have demonstrated an enhanced negative feedback inhibition at the pituitary in PTSD, but have not provided information about central feedback effects, since dexamethasone (DEX) does not penetrate the brain well. The authors therefore examined the change in ACTH and cortisol before and after cortisol administration, which acts at central feedback sites in addition to peripheral targets. METHOD: Blood was obtained from 31 male veterans (18 with PTSD) before, and 8, 40 and 95 min following injection of 17.5 mg cortisol and placebo. RESULTS: A greater decline in ACTH was observed after cortisol injection in PTSD. CONCLUSIONS: Central as well as peripheral negative feedback inhibition may be altered in PTSD. PMID- 16364556 TI - A pilot study of motivation and treatment completion among American Indian adolescents in substance abuse treatment. AB - This study assessed intrinsic and extrinsic motivation toward treatment among American Indian adolescents 13-18 years old admitted to a residential substance abuse treatment program and the relationship of these measures to treatment completion. Participants completed a questionnaire regarding motivation towards treatment and a diagnostic interview. Participant treatment records were also reviewed. Being an older adolescent (16-18 years old), having higher scores on a scale measuring treatment readiness and endorsing a single question regarding concern about legal problems were all associated with treatment completion. The relationship of an additional scale measuring desire for help with substance use problems was mediated by the measure of treatment readiness. This pilot study is the first to assess the relationship of motivation toward substance abuse treatment completion among American Indian adolescents and provides preliminary evidence of the applicability of these concepts to this treatment population. These concepts and their related therapeutic approaches warrant further study in this population. PMID- 16364557 TI - Drinking games in female college students: more than just a game? AB - The present study examined the characteristics of drinking games (DG). Study participants were 164 female college students (Mean age=20; 18-23 years) who completed self-report measures of DG participation and alcohol-related behaviors. Results showed variations in popularity, type of alcohol consumed, and intoxication levels across DG. Findings also revealed that DG involvement (frequency of participation, amount consumed, perceived intoxication, and type of alcoholic beverage consumed) were associated with hazardous alcohol use. Prevention efforts and future research implications are discussed. PMID- 16364558 TI - Cartilage anlagen adapt in response to static deformation. AB - Connective tissue adaptation, including the development of cartilaginous anlagen into bones, is widely believed to be related to dynamic, intermittent load and stress histories. Static stresses, on the other hand, are generally believed deleterious in tissue adaptation. Using serial MRI in a natural human experiment (manipulation and corrective casting of infant clubfoot), we have observed casting produces two effects: (1) the well recognized change in relative positions of the hindfoot anlagen; (2) a newly recognized immediate shape change in the anlagen. These changes seemingly enhance the rate of growth of the anlagen and of the ossific nucleus. The shape change or deformation in the anlagen would occur as a result of alterations in the magnitudes and directions of loading from soft tissue attachments and muscle activity and would necessarily be associated with changes in the stress states within the anlagen and, when present, the ossific nuclei. Given the known role of load and stress history in tissue adaptation, we presume the reduced stress histories influence the enhanced growth rates. These observations contradict some current theories of tissue adaptation since static, rather than dynamic stresses play a crucial role in accelerating the growth and development of anlagen in the infant clubfoot. PMID- 16364559 TI - Meconium aspiration syndrome treatment - new approaches using old drugs. AB - Presently, modern medicine does not offer any disease-modifying treatment for meconium aspiration syndrome (MAS). Several medications with already established safety profiles when employed for similar or other conditions could be useful for MAS treatment. N-Acetylcysteine and DNAse have the capability to reduce viscosity and thickness of meconium by breaking disulfide bonds and slicing DNA, respectively. N-Acetylcysteine, antiprotease drugs, or low pH buffer solutions may have the capability to neutralize meconium's digestive enzymes responsible for lung damage in patients with MAS. All these compounds have great potential to reduce meconium's pathogenic properties which in turn could alleviate MAS severity. PMID- 16364560 TI - Cancer as a chaotic adaptation of the adult stem cell. PMID- 16364561 TI - Avian flu virus H5N1: no proof for existence, pathogenicity, or pandemic potential; non-"H5N1" causation omitted. PMID- 16364562 TI - Salmeterol and paroxetine: more evidence for paradoxical medicine. PMID- 16364563 TI - Perturbations in hypoxia detection: a shared link between hereditary and sporadic tumor formation? AB - The discovery and characterization of the von Hippel Lindau (VHL) syndrome has brought about tremendous advances in understanding the molecular mechanisms of renal cell carcinoma. VHL mutations are known to act through hypoxia inducible factor, which has a physiologic role in detecting hypoxia. Recent investigations into other hereditary forms of kidney cancer with mutations in genes involving energy metabolism and oxidative changes, such as fumarate hydratase, suggest that metabolic changes related to hypoxia detection may be a common mechanism of tumorigenesis. This implicates aberrations in the kidney's physiologic role in detection of hypoxia in tumor formation. Germline mutations of genes involved in energy metabolism and oxidative perturbations lead to tumors in other tissues that detect hypoxia, such as head and neck paragangliomas that occur in the area of the carotid body. Therefore, aberrations in physiologic detection of hypoxia that predispose to tumor formation may not be a mechanism unique to the kidney. Furthermore, inducers of hypoxic perturbations other than germline mutations in metabolic genes may predispose to cancers in organs that have a physiologic role in detecting hypoxia. Conditions that effectively lead to tissue hypoxia in hypoxia detecting tissues is one such mechanism. We propose that some of the common molecular and physiologic mechanisms in heritable forms of kidney cancer, namely detection of hypoxia, may play a role in the genesis of sporadic kidney cancer. We survey evidence suggesting that the mechanism of some recognized risk factors of kidney cancer, such as smoking and obesity, may be due in part to tissue hypoxia, reflecting physiologic detection of hypoxia gone awry. PMID- 16364564 TI - Contextual determinants of temporal control: Behavioral contrast in a free operant psychophysical procedure. AB - The question of how temporal control of responding might be influenced by contingency changes in other contexts was investigated. Each of three pigeons first was exposed to a two-component multiple schedule in which a two-key free operant psychophysical procedure operated in one component and a variable interval schedule operated in the other component. The variable-interval schedule then was changed to extinction while the free-operant psychophysical procedure remained unchanged. Finally, the variable-interval schedule was reintroduced. Response rates on the left key and the estimated temporal threshold under the free-operant psychophysical procedure increased for each pigeon when the alternate component schedule was changed to extinction and then decreased again when the variable-interval schedule was reintroduced. The results suggest one way that temporal control is affected by its context, and may be interpreted through the direct effects of overall reinforcement rate on temporal control mechanisms or the disruptive effects of alternative sources of reinforcement on temporally controlled behavior. PMID- 16364565 TI - The prevalence of drinking and driving in the United States, 2001-2002: results from the national epidemiological survey on alcohol and related conditions. AB - Traffic deaths and injuries are among the most frequent causes of deaths and disability worldwide. In the United States, the National Highway Transportation Safety Administration (NHTSA) reported that approximately 40% of all traffic fatalities were alcohol-related. Yet, information about the prevalence of drinking and driving behaviors of the U.S. general population is lacking. The purpose of this study was to examine the magnitude of driver-based (i.e., driving while drinking and driving after having too much to drink) and passenger-based (i.e., riding with a drinking driver and riding as a passenger while drinking) drinking and driving behaviors confronting contemporary America. The past-year prevalence data were stratified by major sociodemographic characteristics to identify important determinants of drinking and driving behaviors for further research. Data were derived from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC, n=43,093). In 2001-2002 there were 23.4 million, or 11.3%, of American adults who reported engaging in at least one of the four driver- or passenger-based drinking and driving behaviors. The prevalences of passenger-based drinking and driving behaviors were generally greater than those of the driver-based measures. For all four drinking and driving behaviors, age was inversely associated with the risk and males were at greater risk with the associated male-to-female ratios of approximately 3.0. Our data also suggested that Native Americans, individuals who were widowed/separated/divorced or never married, and those with greater than a high school education were also at greater risks of all drinking and driving behaviors. PMID- 16364566 TI - Facilitating entry into drug treatment among injection drug users referred from a needle exchange program: Results from a community-based behavioral intervention trial. AB - We evaluated a case management intervention to increase treatment entry among injecting drug users referred from a needle exchange program (NEP). A randomized trial of a strengths based case management (intervention) versus passive referral (control) was conducted among NEP attenders requesting and receiving referrals to subsidized, publicly funded opiate agonist treatment programs in Baltimore, MD. Logistic regression identified predictors of treatment entry within 7 days, confirmed through treatment program records. Of 247 potential subjects, 245 (99%) participated. HIV prevalence was 19%. Overall, 34% entered treatment within 7 days (intervention: 40% versus control: 26%, p=0.03). In a multivariate "intention to treat" model (i.e., ignoring the amount of case management actually received), those randomized to case management were more likely to enter treatment within 7 days. Additional "as treated" analyses revealed that participants who received 30 min or more of case management within 7 days were 33% more likely to enter treatment and the active ingredient of case management activities was provision of transportation. These findings demonstrate the combined value of offering dedicated treatment referrals from NEP, case management and transportation in facilitating entry into drug abuse treatment. Such initiatives could be implemented at more than 140 needle exchange programs currently operating in the United States. These data also support the need for more accessible programs such as mobile or office-based drug abuse treatment. PMID- 16364567 TI - Pharmacological content of tablets sold as "ecstasy": results from an online testing service. AB - PURPOSE: This study examined the pharmacological content of tablets sold as ecstasy, the variation of tablet content by geographic region, and change in tablet content between 1999 and 2005. METHODS: The sample was comprised of tablets anonymously submitted for laboratory testing between 1999 and 2005 (n=1214). Tablet height, width, geographic region, and year of submission were all used to predict the pharmacological content of the tablets. RESULTS: Overall, 39% of the tablets were comprised of MDMA only, 46% only contained substances other than MDMA and 15% were mixtures of MDMA and other substances. Tablet height and width were inversely related to tablet purity. Ecstasy tablets from California and Florida had decreased likelihoods of containing non-MDMA substances. The purity of tablets decreased over time, which was largely a result of an increasing number of tablets comprised of MDMA along with other substances. CONCLUSIONS: Ecstasy users may be putting themselves at increased risk of substance-induced anxiety, mood, and psychotic disorders by unknowingly ingesting substances other than MDMA. To decrease detrimental health effects, prevention programs should emphasize the impurity of ecstasy tablets and focus on the health impacts of these substances, particularly for populations at high-risk of substance-induced disorders. PMID- 16364568 TI - Longitudinal predictors of injection cessation and subsequent relapse among a cohort of injection drug users in Baltimore, MD, 1988-2000. AB - OBJECTIVE: To determine predictors of injection drug use cessation and subsequent relapse among a cohort of injection drug users (IDUs). METHODS: IDUs in Baltimore, MD were recruited through community outreach in 1988-1989. Among IDUs with at least three follow-up visits, parametric survival models for time to injection cessation (>or=6 months) and subsequent relapse were constructed. RESULTS: Of 1327 IDUs, 94.8% were African American, 77.2% were male, median age was 34 years, and 37.7% were HIV-infected. Among 936 (70.5%) subjects who ceased injection, median time from baseline to cessation was 4.0 years. Three-quarters subsequently resumed injection drug use, among whom median time to relapse was 1.0 year. Factors independently associated with a shorter time to cessation were: age <30 years, stable housing, HIV seropositivity, methadone maintenance treatment, detoxification, abstinence from cigarettes and alcohol, injecting less than daily, not injecting heroin and cocaine together, and not having an IDU sex partner. Factors independently associated with shorter time to injection relapse were male gender, homelessness, HIV seropositivity, use of alcohol, cigarettes, non-injection cocaine, sexual abstinence and having a longer time to the first cessation. CONCLUSIONS: This study provides strong support for targeting cessation efforts among young IDUs and severely dependent, unstably housed, and HIV-infected individuals. PMID- 16364569 TI - Self-Organizing Map (SOM) unveils and visualizes hidden sequence characteristics of a wide range of eukaryote genomes. AB - Novel tools are needed for comprehensive comparisons of interspecies characteristics of massive amounts of genomic sequences currently available. An unsupervised neural network algorithm, Self-Organizing Map (SOM), is an effective tool for clustering and visualizing high-dimensional complex data on a single map. We modified the conventional SOM, on the basis of batch-learning SOM, for genome informatics making the learning process and resulting map independent of the order of data input. We generated the SOMs for tri- and tetranucleotide frequencies in 10- and 100-kb sequence fragments from 38 eukaryotes for which almost complete genome sequences are available. SOM recognized species-specific characteristics (key combinations of oligonucleotide frequencies) in the genomic sequences, permitting species-specific classification of the sequences without any information regarding the species. We also generated the SOM for tetranucleotide frequencies in 1-kb sequence fragments from the human genome and found sequences for four functional categories (5' and 3' UTRs, CDSs and introns) were classified primarily according to the categories. Because the classification and visualization power is very high, SOM is an efficient and powerful tool for extracting a wide range of genome information. PMID- 16364570 TI - Duplication and extensive remodeling shaped POTE family genes encoding proteins containing ankyrin repeat and coiled coil domains. AB - The POTE family genes encode a highly homologous group of primate-specific proteins that contain ankyrin repeats and coiled coil domains. At least 13 paralogous POTE family genes are found on 8 human chromosomes (2, 8, 13, 14, 15, 18, 21 and 22), which can be sorted into 3 groups based on sequence similarity. We identified by a database search a group of additional human ankyrin repeat domain proteins, of which ANKRD26 and ANKRD30A are the best characterized; these are more distant homologs of POTE family proteins. A comprehensive comparison of the genomic organization indicates that ANKRD26 has the genomic structure of the possible ancestor of ANKRD30A and all POTE family genes. Extensive remodeling involving segmental loss and internal duplication appears to have reshaped the ANKRD30A and POTE family genes after the primal duplication of the ancestor gene. We also identified a mouse homolog of human ANKRD26, but failed to find a mouse homolog that bears the structural characteristics of any of the POTE family of proteins. The mouse Ankrd26 may serve as a useful model for the study of the function of human ANKRD26, ANKRD30A and POTE family proteins. PMID- 16364571 TI - Analysis of porcine differential gene expression following challenge with Salmonella enterica serovar Choleraesuis using suppression subtractive hybridization. AB - Swine-adapted Salmonella enterica subsp. enterica serovar Choleraesuis (S. Choleraesuis) is the pathogen most frequently isolated from diseased pigs and may affect host gene expression in a species-specific manner. To characterize the porcine transcriptional response to S. Choleraesuis infection, the mRNA profiles from the mesenteric lymph nodes of three non-infected and three experimentally infected pigs at 24 h post-inoculation were analyzed by suppression subtractive hybridization (SSH). Forty-four up-regulated and 44 down-regulated genes were revealed by differential cDNA screening of 384 forward and 288 reverse subtracted cDNA clones. The DNA sequence of the cDNA clones identified genes with a role in a variety of cellular functions as well as gene products of unknown function. Seven up-regulated genes (CXCL10, CXCR4, SDCBP, DNAJA1, HSPH1, HSP90 and ANXA5) and two functionally related genes (HSP70 and DNAJA4:pDJA1) were selected for further analysis based on their predicted roles in infection and immunity. Real time RT-PCR was performed using RNA collected from a time course of infection spanning from the acute phase (8 h) to the chronic phase (21 days) to confirm and quantitate the up-regulation of the SSH-enriched genes. Correlating with the clinical signs of infection (fever, diarrhea and lethargy), the most dramatic induction of gene expression for all nine genes occurred at 48 h post inoculation. This investigation further defines the porcine response to a host adapted strain of Salmonella by revealing the differential expression of genes with a role in a variety of host cellular functions including innate immunity and cytoskeleton regulation. PMID- 16364572 TI - Umbilical vein plasma concentrations of asymmetrical dimethylarginine are increased in male but not female neonates delivered preterm: a pilot study. AB - BACKGROUND: Infants born term have substantially elevated plasma concentrations of the endogenous nitric oxide synthase antagonist asymmetrical dimethylarginine (ADMA) that normalize with growth. The plasma levels of ADMA in preterm newborns are unknown. SUBJECTS AND METHODS: Plasma concentrations of ADMA, symmetrical dimethylarginine (SDMA) and L-arginine were analyzed from venous umbilical cord blood samples of 19 preterm and 21 term infants by high performance liquid chromatography. RESULTS: Male preterm newborns (n=11) had higher ADMA (median [95% confidence interval (CI)]: 1.90 [1.73-2.10] micromol/l) than females born preterm (n=8; 1.57 [1.24-1.69] micromol/l; p<0.005). In term born males (n=10) and females (n=11) ADMA was significantly lower than in preterm male infants (all p<0.005), and without sex differences. SDMA and L-arginine concentrations were comparable between all groups. ADMA correlated inversely with body weight in male preterm newborns (r=-0.67; p<0.03). CONCLUSION: Male neonates delivered preterm have significantly higher umbilical cord venous plasma concentrations of ADMA compared to female neonates and infants born term. The sex difference and the time course of elevated ADMA may play a role in development and warrant further investigation. PMID- 16364573 TI - Outcome at 2 years for very low birthweight infants in a geographical population: risk factors, cost, and impact of congenital anomalies. AB - AIM: To determine the type and rate of disability at 2 years of age in infants born in the geographically defined population of East Anglia with a birthweight less than 1500 g and to assess the risk factors for disability. STUDY DESIGN: A prospective cohort analysis from all eight neonatal units in East Anglia from 1993-1997 using a single database. METHODS: Local paediatricians assessed children at 2 years using the Health Status Questionnaire and data collection was centrally coordinated. RESULTS: Outcomes for 947 children, 99% of survivors, were available, 74 (7.8%) had severe disability and this was significantly associated with gestational age (p<0.0005), birthweight (p<0.0005) and sex (p=0.046). Major congenital abnormality contributed 27% of all severe disability. The overall cerebral palsy rate was 6.2%, nine children were blind and five had sensorineural hearing loss requiring aids. These children had a high level of use of community services with 19% of the cohort being referred to one or more community service. ELBW infants or those born <30 weeks gestation were 1.5 times and twice as likely to have moderate or severe disability and 2.3 and 5.4 times as likely to have cerebral palsy as those weighing 1000 to 1500 g or >30 weeks gestation. Boys were at higher risk of adverse outcome. CONCLUSIONS: The study was able to define the increased risk associated with being born at lower gestational age or lower birthweight and demonstrates successful ascertainment of outcomes for large local populations at a reasonable cost. PMID- 16364574 TI - A human physiological model describing acetone kinetics in blood and breath during various levels of physical exercise. AB - Physiologically based toxicokinetic (PBTK) modeling of human experimental data suggests difficulties to simultaneously describe the time courses of inhaled polar solvents in blood and breath, especially if exposures occur during physical exercise. We attribute this to the washin-washout effect in the airways. The aim was to develop a PBTK-model that explains the behavior of acetone in blood and exhaled air at different levels of physical exercise. The model includes exchange of inhaled solvent vapor with the blood flow via the mucosa and separate compartments to describe working and resting muscles. The developed model was contrasted to a traditional PBTK-model where the conducting airways were regarded as an inert tube. Our model predictions agrees well with experimentally observed acetone levels in both arterial blood and end- and mixed-exhaled air from 26 inhalation experiments conducted with 18 human volunteers at 0, 50, 100 and 150 W workload. In contrast, the inert-tube model was unable to describe the data. The developed model is to our knowledge the first which explains the toxicokinetics of acetone at such various levels of physical exercise. It may be useful in breath monitoring and to obtain more accurate estimates of absorbed dose during inhalation of polar volatiles. PMID- 16364575 TI - Which progestogen is more likely to increase the risk of fatal myocardial infarction: a combination of epidemiological and trial evidence. AB - OBJECTIVES: To evaluate the impact of metabolic effects of different progestogens on the risk of fatal myocardial infarction is evaluated. METHODS: The changes in (apo)lipoproteins obtained from a randomized trial of three hormone therapy regimens were applied to three models for predicting fatal myocardial infarction derived from the apolipoprotein-related mortality risk (AMORIS) study. In our trial, 487 postmenopausal women were randomized to oral estradiol, with sequential addition of two trimegestone (TMG) doses or norethisterone acetate (NETA), and studied at baseline and after 3, 6, and 12 months. RESULTS: The change from baseline in risk of fatal myocardial infarction, using AMORIS model 3, containing total cholesterol, triglycerides, and apolipoprotein AI, was a 10% reduction for the two TMG doses; NETA had no apparent impact. The differences between treatments were significant at all three time points. When apoB was added in AMORIS model 4, the difference between treatments (5% reduction in the two doses of TMG, compared to NETA) decreased over time, probably due to the effect of dropouts in the NETA group. CONCLUSIONS: This analysis shows different metabolic responses to progestogens in terms of risk of fatal myocardial infarction. Generalization of health benefits or adverse effects seen in trials of hormone therapy to other progestogens could be misleading. PMID- 16364576 TI - Evaluation of sodium alginate as drug release modifier in matrix tablets. AB - Alginates are useful natural polymers suitable for use in the design of pharmaceutical dosage forms. However, the effects of particle size, viscosity and chemical composition of alginates on drug release from alginate matrix tablets are not clearly understood. Hence, 17 grades of sodium alginate with different particle size distributions, viscosities and chemical compositions were used to prepare matrix tablets at various concentrations to screen the factors influencing drug release from such matrices. Particle size was found to have an influence on drug release from these matrices. Sodium alginate was subsequently classified into several size fractions and also cryogenically milled to produce smaller particle size samples. Cryogenic milling could be successfully applied to pulverize coarse alginate particles without changing the quality through degradation or segregation. This study showed the significance of each alginate property in modulating drug release: particle size is important in initial alginic acid gel barrier formation as it affected the extent of burst release; higher alginate viscosity slowed down drug release rate in the buffer phase but enhanced release rate in the acid phase; high M-alginate might be more advantageous than high-G-alginate in sustaining drug release; and, the effect of increasing alginate concentration was greater with larger alginate particles. This can serve as a framework for formulators working with alginates. Furthermore, the results showed that sodium alginate matrices can sustain drug release for at least 8 h, even for a highly water-soluble drug in the presence of a water-soluble excipient. PMID- 16364577 TI - Microdialysis sampling and the clinical determination of topical dermal bioequivalence. AB - Our objective was to determine whether the degree of variability associated with dermal microdialysis allows its practical application to determinations of bioequivalence of topically applied agents with a reasonable number of subjects. A statistical review of literature data was conducted to estimate the variances associated with subject-to-subject variability and the probe-to-probe variability within the subjects. In order to successfully utilise dermal microdialysis to establish bioequivalence of topically applied agents, particular care must be applied to study design. Due to the inherent variability between subjects, to maintain subject numbers at reasonable levels, each subject should act as their own control, thus removing the element of subject-to-subject variability from calculations of sample sizes. It is also recommended that measurements are made in duplicate in each subject to reduce the element of variability further. It is then possible to demonstrate, within 80-125% confidence limits and a subject population of approximately 20, that two formulations are bioequivalent. PMID- 16364578 TI - Development of pH-sensitive liposomes that efficiently retain encapsulated doxorubicin (DXR) in blood. AB - We have reported that targeted, pH-sensitive sterically stabilized liposomes are able to increase the cytotoxicity of DXR in vitro against B lymphoma cells, but the rate of release of DXR in plasma was too rapid to permit the results to be extended to in vivo applications. The purpose of the study reported here is two fold. First, to understand the mechanism of the rapid release of DXR from pH sensitive sterically stabilized liposomes (PSL) in human plasma. Second, to reformulate the above liposomes to improve their drug retention, while retaining their pH sensitivity. The stability of the PSL formulations in human plasma was evaluated by comparing the rate of release of encapsulated DXR with that of HPTS, a water-soluble fluorescent marker. Since DXR, but not HPTS, a water soluble-less membrane permeable fluorescence marker, was rapidly released from liposomes in the presence of plasma, the rapid release of DXR is likely caused by the diffusion of DXR molecules through the lipid bilayer, not by the disruption of the membrane. In order to develop more stable PSL formulations, various molar ratios of the membrane rigidifying lipid, hydrogenated soy HSPC and/or CHOL, were added to the lipid composition and the rate of release of encapsulated solutes and pH-sensitivity were evaluated. The compositions that showed the best drug retention and pH-sensitivity were a mixture of DOPE/HSPC/CHEMS/CHOL/mPEG(2000) DSPE at a molar ratio of 4:2:2:2:0.3 and DOPE/HSPC/CHEMS/CHOL at a molar ratio of 4:2:2:2. Our formulations, if targeted to internalizing antigens on cancer cells, may increase intracellular drug release rates within acidic compartment, resulting in a further increase in the therapeutic efficacy of targeted anticancer drug-containing liposomes. PMID- 16364580 TI - Changes in otoacoustic emissions and high-frequency hearing thresholds in children and adolescents. AB - With the aim of characterizing the loss of high frequency hearing sensitivity in children, hearing thresholds and otoacoustic emissions were measured in a group of 126 normal hearing children and adolescents aged from 6 to 25 years. The subjects were divided into four 5-year age groups. Hearing thresholds over a range of 125 Hz-12.5 kHz were similar in all age groups, the average hearing threshold at 16 kHz was significantly elevated in the oldest age group. The response values of transiently evoked otoacoustic emissions (TEOAEs) significantly declined with age; the decline was negatively correlated with the hearing loss at 16 kHz. Significantly larger TEOAE responses and average distortion-product otoacoustic emission (DPOAE) values at 6.3 kHz were present in the youngest group in comparison with the other three older groups. Spontaneous otoacoustic emissions (SOAEs) were present in 70.8% of the children (in either one or both ears) with the greatest prevalence in the 11-20-year-old subjects. In the 21-25-year-old group, the hearing loss at 16 kHz was significantly smaller in ears with SOAEs than in ears without SOAEs. The results demonstrate that the increase in the high frequency hearing threshold at 16 kHz, which starts at ages over 20 years, is correlated with a decrease in the TEOAE responses at middle frequencies. PMID- 16364581 TI - Insights into elevated distortion product otoacoustic emissions in sickle cell disease: comparisons of hydroxyurea-treated and non-treated young children. AB - Distortion product otoacoustic emissions (DPOAEs) were examined in 15 normal hearing African-American children between the ages of 6 and 14 years with homozygous sickle cell disease (SCD), who were on a regimen of hydroxyurea (HDU), a drug that reduces inflammatory processes and symptoms of SCD; a matched group of 15 African-American children with homozygous SCD not on HDU; and 15 African American children with normal hemoglobin. DPOAEs were evoked by 13 primary tone pairs with f2 frequencies ranging from 1000 to 4500 Hz. Increased DPOAE amplitudes, believed to be a precursor of eventual hearing loss, were evident in children with SCD who were not receiving HDU. Those taking HDU had DPOAE amplitudes similar to normal controls. These findings suggest that HDU, in addition to reducing symptoms of SCD, may play a role in inhibiting or preventing cochlear pathology and hearing loss in individuals with SCD. PMID- 16364582 TI - Representation of cerebral bridging veins in infants by postmortem computed tomography. AB - The postmortem diagnosis of shaken baby syndrome, a severe form of child abuse, may be difficult, especially when no other visible signs of significant trauma are obvious. An important finding in shaken baby syndrome is subdural haemorrhage, typically originating from ruptured cerebral bridging veins. Since these are difficult to detect at autopsy, we have developed a special postmortem computed tomographic (PMCT) method to demonstrate the intracranial vein system in infants. This method is minimally invasive and can be carried out conveniently and quickly on clinical computed tomography (CT) systems. Firstly, a precontrast CT is made of the infant's head, to document the original state. Secondly, contrast fluid is injected manually via fontanel puncture into the superior sagittal sinus, followed by a repeat CT scan. This allows the depiction of even very small vessels of the deep and superficial cerebral veins, especially the bridging veins, without damaging them. Ruptures appear as extravasation of contrast medium, which helps to locate them at autopsy and examine them histologically, whenever necessary. PMID- 16364583 TI - MR-guided intervention in women with a family history of breast cancer. AB - OBJECTIVE: A study was undertaken to assess the clinical value of magnetic resonance (MR) imaging-guided interventions in women with a family history, but no personal history of breast cancer. METHODS AND PATIENTS: Retrospective review was performed on 63 consecutive women who had a family history, but no personal history of breast cancer. A total of 97 lesions were referred for an MR-guided intervention. Standardized MR examinations (1.0 T, T1-weighted 3D FLASH, 0.15 mmolGd-DTPA/kg body weight, prone position) were performed using a dedicated system which allows vacuum assisted breast biopsy or wire localization. RESULTS: Histologic findings in 87 procedures revealed 9 (10%) invasive carcinomas, 12 (14%) ductal carcinomas in situ, 2 atypical ductal hyperplasias (2.5%) and 2 atypical lobular hyperplasias (2.5%). Sixty-two (71%) benign histologic results are verified by an MR-guided intervention, retrospective correlation of imaging and histology and by subsequent follow-up. In ten lesions the indication dropped since the enhancing lesion was no longer visible. Absent enhancement was confirmed by short-term re-imaging of the noncompressed breast and by follow-up. CONCLUSION: Malignancy was found in 24%, high-risk lesions in 5% of successfully performed MR-guided biopsy procedures. A 57% of MR-detected malignancies were ductal carcinoma in situ. In 10% of the lesions the intervention was not performed, since no enhancing lesion could be reproduced at the date of anticipated intervention. Such problems may be avoided if the initial MRI is performed in the appropriate phase of the menstrual cycle and without hormonal replacement therapy. PMID- 16364584 TI - Simultaneous determination of amoxicillin and ranitidine in rat plasma by high performance liquid chromatography. AB - A high-performance liquid chromatography method using ultraviolet detection at 230 nm for the simultaneous determination of amoxicillin and ranitidine in rat plasma has been validated. Plasma samples after pretreatment with acetonitrile to effect deproteinization were dried under N2 and reconstituted with water. The standard calibration curves for amoxicillin and ranitidine were linear (r2=0.9999) over the concentration range of 0.2-20 microg ml-1 and 0.03-6 microg ml-1 in rat plasma, respectively. The intra- and inter-day assay variability range for amoxicillin was 2.4-8.5% and 3.2-11.7%, and for ranitidine was 1.7-9.0% and 4.5-10.1%, respectively. This method has been successfully applied to a pharmacokinetic study after oral coadministration of amoxicillin and ranitidine to rats. PMID- 16364585 TI - Evaluation of the chemiluminescent nitrogen detector for solubility determinations to support drug discovery. AB - Solubility measurements using chemiluminescent nitrogen detection (CLND) has advantages of reduced compound requirement and increased throughput compared to UV-spectrophotometric and HPLC-based measurements. CLND with direct flow injection was evaluated for the measurement of thermodynamic solubility to support drug discovery. The limit of quantitation (LOQ), accuracy, and day-to-day reproducibility of the detector were measured. Measurements made on CLND were compared to those obtained from UV spectrophotometry and HPLC. Based on the results obtained, it was concluded that the CLND performs satisfactorily for discovery stage thermodynamic solubility measurements. PMID- 16364586 TI - The application of inductively coupled plasma mass spectrometry in pharmaceutical and biomedical analysis. AB - With the development of life science, pharmaceutical and biomedical analysis becomes more and more important in medical science. Further studies will be hopefully established if it is possible to use inorganic elemental standards or small organic compounds in the quantitative determination of all kinds of drugs, nucleotides and sulfur or phosphorus containing peptides and proteins at appropriate concentration with an acceptable accuracy. Since 1980, inductively coupled plasma mass spectrometry (ICP-MS) has emerged as a new and powerful analytical technique which is suitable for element and isotope analysis. It offers extremely wide detection range of element and co-analysis of most elements in the periodic table. Also, it can be applied to perform qualitative, semiquantitative, and quantitative analysis and isotopic ratios through mass-to electric charge ratio. With the help of ICP-MS, the struggle of searching for an excellent quantification technique in, e.g. drugs and proteomics has come appreciably close to an end. This review mainly focuses on the introduction of application of ICP-MS in pharmaceutical and biomedical analysis. Some problems in application and the handling strategies are simply presented at the end. PMID- 16364587 TI - Urea as an endogenous surrogate in human microdialysis to determine relative recovery of drugs: analytics and applications. AB - During in vivo microdialysis studies time-consuming and laborious bedside calibration methods, e.g. retrodialysis, have to be performed. To reduce the burden on the patient it would be desirable to establish a reliable, time-saving calibration technique to obtain the in vivo recovery describing the relative drug transfer across the membrane of the microdialysis probe. The performed study aimed to evaluate and validate the use of urea as an endogenous reference compound to determine relative in vivo recovery of anti-infectives, e.g. linezolid used herein as model drug. In order to meet the special requirements imposed by microdialysis to measure urea concentrations in very small sample volumes ( approximately 10 microL) a photometric assay in 96-well microtiter plates was established based on the method of Berthelot. Subsequently, concentration- and flow rate-dependence were evaluated in vitro to determine the relative recovery (RR) of urea. Finally, urea and linezolid concentrations in human microdialysis samples were measured. The developed assay was validated according to international guidelines and met all requirements. Relative in vitro recovery was found to be independent from concentration and dependent on flow rate. Subsequently, relative in vivo recovery of urea was correlated with relative in vivo recovery of linezolid obtained by the traditional retrodialysis method. In healthy volunteers, the mean ratio of the relative recovery of linezolid to the relative recovery of urea was 0.6 for the subcutaneous (s.c.: CV 33.4%, n = 48) and 0.7 for the intramuscular probe (i.m.: CV 18.8%, n = 40), respectively. In critically ill patients this ratio was 0.7 for both tissues (s.c.: CV 32.8%, n = 18; i.m.: CV 22.1%, n = 17). Successful calibration of the urea reference technique without the need to use in vitro data will further promote the application of microdialysis in clinical studies especially in critically ill patients, as it reduces the imposed burden to a minimum. PMID- 16364589 TI - Hepatic resection for malignant melanoma. AB - INTRODUCTION: Patients with metastatic melanoma have a poor prognosis with a median survival of approximately 6 months. There is a paucity of data regarding the natural history of the disease in the small subset of patients who are suitable for hepatic resection for metastatic disease confined to the liver. PATIENTS AND METHODS: Five patients were identified from a prospectively collected database of over 1000 liver resections performed in Basingstoke between 1986 and 2004. Clinical details and survival were analysed. RESULTS: Two patients died within 12 months of resection from extrahepatic disease. The other three patients are alive and disease-free at 76, 92 and 147 months, respectively. DISCUSSION: In carefully selected cases, hepatic resection for metastatic melanoma to the liver can result in long-term survival. Criteria for selection of patients as suitable for resection remain unclear. PMID- 16364588 TI - Separation of nicotinic acid and its structural isomers using 1-ethyl-3 methylimidazolium ionic liquid as a buffer additive by capillary electrophoresis. AB - The growing interest in application of ionic liquids (ILs) in analytical chemistry has been observed. The aim of presented investigation was to verify whether ILs would be a suitable modifier of the background electrolyte (BGE) for pharmaceutical analysis of the closely related drug analogues. The study demonstrates the use of 1-ethyl-3-methylimidazolium tetrafluoroborate (1E-3MI TFB) ionic liquid as modifiers in the separation of nicotinic acid and its structural isomers by capillary electrophoresis. Dependences of the ionic liquid concentration in a BGE on the separation parameters like migration time, resolution factor and width at peak's baseline have been compared. The separation mechanism involves the free imidazolium ions, which can interact with inner surface of the capillary wall. Increased 1E-3MI-TFB concentration to 150 mmol/L caused decrease of migration times of analytes, improve peaks shape and increase of separation performances. At this ionic liquid concentration in a BGE resolution factor between nicotinic and isonicotinic acids increased to 1.86. The proposed CE separation procedure is highly reproducible and can be applied in qualitative and quantitative analysis of carboxylic acids. PMID- 16364590 TI - [Practical anatomy]. PMID- 16364591 TI - Mothballs and obstetric anesthesia. PMID- 16364592 TI - [Block of the lumbar plexus and its branches]. PMID- 16364593 TI - Modeling of myocardial contractility using parameterized super-quadric SPECT images. AB - We developed methods to represent cardiac motility. Using an innovative model, we estimated several parameters of cardiac features. We implemented the parameterized super quadric model to visualize the motion of a left ventricle (LV) with OpenGL and Visual C++. We displayed myocardial wall thickening with a super-ellipsoidal model. The time frames in this model changed the measured thickening count. We also parameterized motility using the parameterized super quadric model. We analyzed the motility of the LV myocardium and tested its criteria using a validation study of seven normal subjects and 26 patients with prior myocardial infarction. To analyze motility, we used mean and variance of total motion during a cardiac cycle. The average of a normal subject was 0.46 and variance was 0.02. For patients, average and variance of motility were 0.59 and 0.08 respectively. Although the average value did not differ between normal subjects and patients, the variance differed significantly. Thus, we were able to estimate the difference between normal subjects and patients. In patients, motility was 128% higher than in normal subjects, and the variance was 328% higher. In the patient study, quantity of motion decreased rapidly in a stressed state. The visualization for contractility displayed 15 segment variables; we were able to rotate the locations of all points with a mouse interface. We were able to visualize most of the factors for cardiac motility and cardiac features. We expect that this model can distinguish between normal subjects and abnormal subjects, and that we can produce an exact analysis of momentum using this model. PMID- 16364594 TI - The role of intraoperative ultrasonography in zygomatic complex fracture repair. AB - Intraoperative assessment of the zygomatic arch is very important in achieving adequate repositioning. The correct alignment of the zygomatic arch indicates the proper position of the zygomatic bone and ensures adequate prominence of the lateral midfacial aspect. The aim of this study was to estimate the value of ultrasonography as an intraoperative repositioning control. In a clinical study of 25 patients, ultrasonography was employed for intraoperative visualization of the zygomatic arch before and after fracture repositioning. Twelve patients presented with isolated zygomatic arch fractures and 13 with combined fractures of the zygomatic bone and arch. The ultrasonographic findings were compared to the radiological and clinical findings. Ultrasonography was able to detect all fractures and dislocations of the zygomatic arch. It was possible to assess the repositioning in 24 out of 25 cases using ultrasonography. The ultrasound images were concordant with the radiographs. Clinical assessment by palpation only succeeded in isolated zygomatic arch fractures with an m-shaped impression, whereas it remained uncertain in nearly all cases with a different dislocation pattern. Ultrasonography was rapid and easy to perform, and is recommended as an intraoperative visualizing tool in all midfacial fractures with displacement of the zygomatic arch. PMID- 16364595 TI - A prospective comparison of octyl-2-cyanoacrylate and suture in standardized facial wounds. AB - Regarding the cosmetic results of wound closure using tissue adhesives as opposed to sutured wounds, most published studies are performed on children and with non standardized lacerations, which makes it difficult to transfer the results to facial incisional wounds in adults. There are also conflicting conclusions about the cosmetic results. The purpose of this randomized prospective study was to compare the cosmetic outcomes of wound closure with sutures and octyl-2 cyanoacrylate in standardized facial wounds in adults. To compare very similar facial wounds, the infraorbital cut (lower eyelid incision) was used in 45 patients. The assessment was performed by patients and surgeons blind to the data. Phase-shifting profilometry was used to measure the profile of the scars. No early complications were observed in any group. The wound depth in the tissue adhesive group was significantly greater than in the suture group. There were no differences in the overall cosmetic results among all patients in the two groups. Interestingly, in the younger patients the result of the sutured wounds was superior to that of the adhesive-treated patients. The Dermabond skin adhesive provides a means of closure of facial surgical wounds without early complications, but adjustment of the edges of the wound and the cosmetic result in younger patients are less successful than with thin sutures. PMID- 16364596 TI - Dyslexia as a possible aspect of neurocognitive impairment in schizophrenia. PMID- 16364597 TI - Putative association of the carboxy-terminal PDZ ligand of neuronal nitric oxide synthase gene (CAPON) with schizophrenia in a Colombian population. PMID- 16364598 TI - Self-reported mental health difficulties and subsequent risk for schizophrenia in females: a 5-year follow-up cohort study. AB - BACKGROUND: Patients with schizophrenia often report a history of premorbid mild to severe psychological distress. We investigated the association between self reported mental health difficulties and later psychiatric hospitalization for schizophrenia. METHODS: 13,357 females aged 17, mandatory assessed by the Israeli Draft Board were followed up over 5 years for psychiatric hospitalization by means of the Israeli National Psychiatric Hospitalization Case Registry. Seventeen females, judged healthy at Draft Board assessment, were hospitalized for schizophrenia or schizoaffective disorder over the follow-up period. RESULTS: There was a significant monotonic association between increasing self-reported mental health difficulties (psychological distress and increasing need for psychological counseling) and prevalence of schizophrenia [odds ratios over four levels: 1.56; 95% CI:1.04 to 2.34; chi2 (1) = 4.62, p = 0.03], after controlling for low IQ, immigration, SES, and presence of psychiatric disorders at age 17. Increasing severity of self-reported mental health difficulties was related to earlier age of first hospitalization [r = -0.48, p = 0.05]. CONCLUSIONS: Increased undifferentiated self-reported mental health difficulties are associated with increased risk of later hospitalization for schizophrenia prior to age 23 in females. This may reflect the prodromal phase of the illness. PMID- 16364599 TI - Enzyme-linked immunosorbent assay using bacterial recombinant proteins of human BP230 as a diagnostic tool for bullous pemphigoid. AB - BACKGROUND: By immunoblot analyses of normal human epidermal extracts, the 230kDa bullous pemphigoid antigen (BP230) is recognized by most bullous pemphigoid (BP) sera. We produced different recombinant glutathione-S-transferase-fusion proteins, which roughly presented N-terminal domain, central rod domain and C terminal domain of human BP230. OBJECTIVE: In the present study, we developed an enzyme-linked immunosorbent assay (ELISA) using the recombinant proteins for detection of anti-BP230 IgG antibodies and assessed the usefulness of this assay in conjunction with an anti-BP180 ELISA to establish the diagnosis of BP. METHODS: Using the bacterial recombinant proteins of N-terminal and C-terminal domains, we developed an ELISA. A receiver-operating-characteristic (ROC) analysis was performed to determine a cut-off value for the BP230 ELISA. RESULTS: By this BP230 ELISA, 173 (72.4%) of 239 BP sera were positive, while only one (1.1%) of 94 sera from pemphigus vulgaris and pemphigus foliaceus patients was positive and all the 109 normal control sera were negative. Thus, the sensitivity and specificity of the BP230 ELISA were 72.4 and 99.5%, respectively. Interestingly, while 54 (84.4%) of 64 BP sera in active stage and 113 (64.6%) of 175 BP sera in remission were positive in BP180 ELISA, 37 (57.8%) of 64 BP sera in active stage and 136 (77.7%) of 175 BP sera in remission were positive in BP230 ELISA. These results indicate that the titer of anti-BP230 antibodies is not related with disease activity in some BP cases. Most significantly, by combining the results of BP230 ELISA and BP180 ELISA, 232 (97.1%) of 239 BP sera were positive. CONCLUSION: The combination of BP230 ELISA and BP180 ELISA is the highly sensitive method for the diagnosis of BP. PMID- 16364600 TI - A unique monoclonal antibody 29A stains the cytoplasm of amniotic epithelia and cutaneous basement membrane. AB - BACKGROUND: The basic function of epithelia is to provide a boundary between tissue and its external environment, and is achieved by a wide variety of components including extracellular molecules. Multiple monoclonal antibodies raised against epithelial antigens have helped identify a range of distinct, novel protein epitopes. OBJECT: In this study, we raised a monoclonal antibody to detect a novel epithelial molecular component. METHODS: We have produced a mouse monoclonal antibody using normal human amniotic tissue as an immunogen. The monoclonal antibody was subsequently immunohistochemically screened, and the target antigen was cloned using an immunoscreening method. RESULT: In the course of the screening, we identified unique antibody staining patterns within the cytoplasm of a subset of amniotic cells at intervals within the normal placental epithelia. By immunoscreening, we identified this candidate gene as laminin receptor (LR). By dot blot analysis, this antibody reacted with recombinant LR. The same localization of the antigen and LR was proved by a double staining immunofluorescence test in the placenta. This monoclonal antibody unexpectedly demonstrated linear staining within the dermal-epidermal junction of normal human skin but failed to react within the keratinocyte cytoplasm. CONCLUSION: We have produced and characterized a novel monoclonal antibody 29A that recognizes an LR related molecule, which demonstrated a unique staining pattern. This monoclonal antibody might be a useful tool for further investigations into the epithelial tissues and the cutaneous basement membrane (BM). PMID- 16364601 TI - Genotyping of Candida albicans on the basis of polymorphisms of ALT repeats in the repetitive sequence (RPS). AB - BACKGROUND: Candida albicans is one of the most important etiologic agents causing superficial and deep fungal infections. For prevention of candidiasis, it is important to develop a rapid system that discriminates C. albicans at the strain level. OBJECTIVE: To develop a system that can identify C. albicans at the strain level. METHODS: Genomic DNAs were purified from 179 clinical isolates of C. albicans, and were used as templates for PCR amplification of 25S rDNA and ALT repeats in repetitive sequences (RPSs). PCR products generated from ALT repeats were digested with EcoRI and/or ClaI in order to study the relationships between restriction profiles and amplification profiles. RESULTS: One hundred and seventy nine clinical isolates were grouped into genotypes A (92 isolates), B (38 isolates) and C (49 isolates) on the basis of their 25S rDNA, and each was further classified into five types (types 3, 4, 3/4, 2/3/4 and 3/4/5) by PCR amplification targeting ALT repeats. Type 3 C. albicans constituted the majority of isolates in any genotypes (66.3% for genotype A, 76.3% for genotype B and 73.4% for genotype C). Each C. albicans type showed several amplification patterns, indicating the existence of subtypes. RFLP analysis revealed that restriction profiles of PCR products corresponded to amplification patterns from PCR. CONCLUSION: The present results indicate that PCR amplifications targeting 25S rDNA and ALT repeats are useful for rapid genotyping and distinction of C. albicans involved in superficial candidiasis. PMID- 16364602 TI - Evaluation of the Bactec microbial detection system for culturing miscellaneous sterile body fluids. AB - This study sought to evaluate the efficacy of the Bactec blood culture system for culturing sterile body fluids, in comparison with traditional culture methods. A total of 906 specimens were cultured using both conventional media and the Bactec detection system. Differences in numbers of isolates determined by each system were compared. Clinically significant microorganisms were isolated from 15.3% (139) of 906 specimens by both the Bactec system and conventional culture, whereas for 8.8% (80) of the specimens a positive culture was obtained by Bactec only. Of the 80 specimens in which growth was detected only in the Bactec blood culture system, 11.0% were comprised of cerebrospinal fluid, 9.9% peritoneal fluid, 4.9% pleural fluid, 4.5% synovial fluid and 9.1% bone marrow materials. Strains of Brucella melitensis, Neisseria meningitidis, Pseudomonas fluorescens and Rothia dentocariosa were cultured only by the Bactec system. In conclusion, the Bactec blood culture system might be advantageous for isolation of fastidious microorganisms such as Brucella, especially from cerebrospinal and peritoneal fluid specimens. PMID- 16364603 TI - Isolation and characterization of a 30 kD antifungal protein from seeds of Sorghum bicolor. AB - An antifungal protein of about 30,000 Da was isolated from seeds of Sorghum bicolor L. using chromatographic techniques, including gel filtration, ion exchange, and high-performance liquid chromatography in a reverse-phase column. This protein (termed 30 kD protein) showed a minimal inhibitory concentration of 36 microg/ml for Candida parapsilosis and C. tropicalis, and 18 microg/ml for C. albicans. The 30 kD protein inhibited adherence to the cover glass and formation of a germinative tube of C. albicans at concentrations over 300 and 150 microg/ml, respectively. Transmission electron microscopy of yeast forms of C. albicans after incubation with 18 mug/ml of the 30 kD protein for 24 h revealed marked ultrastructural changes in the fungus. No toxicity of the 30 kD protein to the culture of Hep2 cells at concentrations equal to or less than 1,000 microg/ml was observed. PMID- 16364604 TI - Transmembrane domain prediction and consensus sequence identification of the oligopeptide transport family. AB - Few polytopic membrane proteins have had their topology determined experimentally. Often, researchers turn to an algorithm to predict where the transmembrane domains might lie. Here we use a consensus method, using six different transmembrane domain prediction algorithms on six members of the oligopeptide transport family, all of which have been experimentally characterized. PSI-BLAST results indicate that the six chosen oligopeptide transport family members are distributed throughout most branches of the phylogram, suggesting that these members represent a broad view of the oligopeptide transport family. We combined the prediction algorithms with a multiple sequence alignment, and consensus transmembrane domains were assigned not only based on algorithmic output, but also based on conserved familial motifs found by analysis of the PSI-BLAST results. The consensus method combined with the "charge-difference rule" yields a model topology for the family containing 12 transmembrane domains with the N- and C-termini facing extracellular. PMID- 16364605 TI - Isolation of Mycobacterium septicum from the sputum of a patient suffering from hemoptoic pneumonia. AB - A rapidly growing mycobacterium was isolated in pure culture from the sputum of a 78-year-old woman suffering from hemoptoic pneumonia. The isolate exhibited an antimicrobial susceptibility pattern and a biochemical profile similar to that of Mycobacterium septicum ATCC 700731(T) and shared 100% 16S rRNA gene sequence similarity with this type strain over 1480 bp. Its partial rpoB sequence shared 98.2% similarity with the latter species, suggesting the isolate was representative of a sequevar of M. septicum. Contrary to the type strain, this isolate was resistant to azithromycin, erythromycin, vancomycin and tobramycin. This case illustrates the first isolation from sputum of M. septicum, a member of the Mycobacterium fortuitum group previously isolated once from bacteremia. Strain D13 has been deposited in the Collection de l'Institut Pasteur as CIP 108512. PMID- 16364606 TI - Survey of environmental biocontamination on board the International Space Station. AB - The International Space Station (ISS) is an orbital living and working environment extending from the original Zarya control module built in 1998. The expected life span of the completed station is around 10 years and during this period it will be constantly manned. It is inevitable that the ISS will also be home to an unknown number of microorganisms. This survey reports on microbiological contamination in potable water, air, and on surfaces inside the ISS. The viable counts in potable water did not exceed 1.0 x 10(2) CFU/ml. Sphingomonas sp. and Methylobacterium sp. were identified as the dominant genera. Molecular analysis demonstrated the presence of nucleic acids belonging to various pathogens, but no viable pathogens were recovered. More than 500 samples were collected at different locations over a period of 6 years to characterize air and surface contamination in the ISS. Concentrations of airborne bacteria and fungi were lower than 7.1 x 10(2) and 4.4 x 10(1) CFU/m3, respectively. Staphylococcus sp. was by far the most dominant airborne bacterial genus, whereas Aspergillus sp. and Penicillium sp. dominated the fungal population. The bacterial concentrations in surface samples fluctuated from 2.5 x 10(1) to 4.3 x 10(4) CFU/100 cm2. Staphylococcus sp. dominated in all of these samples. The number of fungi varied between 2.5 x 10(1) and 3.0 x 10(5) CFU/100 cm2, with Aspergillus sp. and Cladosporium sp. as the most dominant genera. Furthermore, the investigations identified the presence of several (opportunistic) pathogens and strains involved in the biodegradation of structural materials. PMID- 16364607 TI - A rationally assembled microbial community for growing Tagetes patula L. in a lunar greenhouse. AB - Well-defined plant-associated bacteria were used for growing French marigolds (Tagetes patula L.) in anorthosite, a substrate of low bioavailability, analogous to a lunar rock. The consortium was composed mainly of plant growth-promoting rhizobacteria and biocontrol agents that were used for seed inoculation. Simultaneously, the sterile substrate was inoculated with the siliceous bacterium Paenibacillus sp. IMBG156. The plant benefited from bacterial activity which resulted in stimulation of seed germination, better plant development, and finally in flowering of inoculated tagetes. In contrast, control plants grew poorly in sterile anorthosite and never flowered. Analysis of bacterial community composition showed that both species colonized plant roots, and there were no shifts in the consortium structure in the rhizosphere of French marigolds within 6 weeks. Paenibacillus sp. IMBG156 was able to release some elements (Ca, Fe, Si) from substrate anorthosite. It was assumed that a rationally assembled consortium of bacterial strains supported growth and development of the model plant under growth-limiting conditions, at least by means of bioleaching and delivering of essential nutritional elements to the plants, and by promoting plant growth. PMID- 16364608 TI - Surviving inside a macrophage: the many ways of Brucella. AB - Bacteria of the genus Brucella are intracellular pathogens capable of survival and replication within macrophages of mammalian hosts. Recent advances have shed light on virulence factors and host functions involved at various stages of the Brucella intracellular life cycle. This review focuses on how this pathogen uses multiple strategies to circumvent macrophage defense mechanisms and generate an organelle permissive for replication. PMID- 16364609 TI - Combination therapy using minocycline and coenzyme Q10 in R6/2 transgenic Huntington's disease mice. AB - Huntington's disease (HD) is a fatal neurodegenerative disorder of genetic origin with no known therapeutic intervention that can slow or halt disease progression. Transgenic murine models of HD have significantly improved the ability to assess potential therapeutic strategies. The R6/2 murine model of HD, which recapitulates many aspects of human HD, has been used extensively in pre-clinical HD therapeutic treatment trials. Of several potential therapeutic candidates, both minocycline and coenzyme Q10 (CoQ10) have been demonstrated to provide significant improvement in the R6/2 mouse. Given the specific cellular targets of each compound, and the broad array of abnormalities thought to underlie HD, we sought to assess the effects of combined minocycline and CoQ10 treatment in the R6/2 mouse. Combined minocycline and CoQ10 therapy provided an enhanced beneficial effect, ameliorating behavioral and neuropathological alterations in the R6/2 mouse. Minocycline and CoQ10 treatment significantly extended survival and improved rotarod performance to a greater degree than either minocycline or CoQ10 alone. In addition, combined minocycline and CoQ10 treatment attenuated gross brain atrophy, striatal neuron atrophy, and huntingtin aggregation in the R6/2 mice relative to individual treatment. These data suggest that combined minocycline and CoQ10 treatment may offer therapeutic benefit to patients suffering from HD. PMID- 16364610 TI - Spectroscopic and catalytic studies of lipases in ternary hexane-1-propanol-water surfactantless microemulsion systems. AB - A series of water-in-oil microemulsion systems formulated without surfactant were used to solubilize lipases from Rhizomucor miehei and Candida antarctica B. The effect of the system's composition on the velocity of enzymic reactions was investigated following a model esterification reaction. The interaction between enzymes and the microemulsion environment was studied by steady state fluorescence spectroscopy. The site of localization of the enzyme within the different microdomains of the dispersed phase was investigated by applying the fluorescence energy transfer technique. To determine the properties of the interface between water and organic solvent of the surfactantless microemulsion systems the Electron Paramagnetic Resonance (EPR) spectroscopic technique was applied. The results indicated that even at low water content, water-rich structures are formed. This was confirmed by conductivity measurements. By the addition of enzyme it was observed that when the aqueous phase of the surfactantless microemulsion systems exceeds 2% (v/v) the enzyme retains its catalytic activity, as it is located within the water pools that protect it from the organic solvent. These confined water phases show a propanol rich interface with hexane and their structure depends on the system's composition. PMID- 16364611 TI - The role of transporters in drug interactions. AB - Transport proteins play an important role in the adsorption, distribution and elimination of a wide variety of drugs. Therefore, it is not surprising that transporter-based drug interactions can occur in the clinic. These interactions can lead to changes in toxicity and/or efficacy of the affected drug. Here, we review such interactions and ask if these interactions could have been predicted from in vitro data. Conducting such in vitro-in vivo correlation is important for predicting future transporter-based drug interactions. PMID- 16364612 TI - Development and evaluation of an in vitro method for prediction of human drug absorption II. Demonstration of the method suitability. AB - The aim of the present work was to assess the actual suitability and general applicability of a new in vitro permeation method based on an appositely developed artificial membrane to evaluate and predict drug absorption potential. The proposed method was employed to evaluate the apparent permeability of a set of 21 structurally diverse drugs having very different solubility and permeability properties, covering the whole range of fraction absorbed in humans (F(a) from 13 to 100%); 13 of the drugs in this study were part of the list suggested by FDA for validation of in vitro permeation methods. An excellent linear correlation (R(2)=0.957) was obtained between artificial membrane apparent permeability and human absorption data in the whole range of F(a) values examined (including all the drugs belonging to the above FDA list), indicating the good predictive ability of the proposed method not only for highly absorbed hydrophobic compounds but also, differently from other in vitro permeation methods, for poorly or middling permeable drugs. The predictive ability of the new method was greater than those obtained for the same set of drugs with PAMPA and Caco-2 permeability literature data, probably due to the poor sensitivity of these methods towards hydrophilic drugs. The better performance of our artificial membrane was attributed to the hydrophilic nature of the support that, differently from the commonly used hydrophobic supports, offers less resistance to permeation of hydrophilic compounds. A comparison of permeation data of theophylline, ketoprofen, aciclovir and furosemide (selected, respectively, as models of I-IV BCS classes) obtained using a Caco-2 cell based dynamic method and the developed artificial membrane and the corresponding F(a) values in humans further confirmed the suitability of the proposed permeation method as predictor of the oral absorption of passively absorbed drugs. PMID- 16364613 TI - [Quality of life in HIV-infected children and adolescents under highly active antiretroviral therapy: change over time, effects of age and familial context]. AB - OBJECTIVE: To evaluate Quality of life (QoL) of HIV-infected children under highly active antiretroviral therapies, and its change over 18 months. MATERIALS AND METHODS: QoL was evaluated by self-administred questionnaires (french versions of AUQUEI, OK-ado, and compilation of both) in 23 young living with HIV/AIDS (6-15 yrs), under antiretroviral multitherapies, and re-evaluated 18 months later in 19 of them. RESULTS: At baseline, QoL in HIV-infected children adolescents was relatively good. The answers given to each items and the mean score from infected children were similar to those obtained in uninfected healthy children. Moreover, infected adolescents distinguished definitly from healthy adolescents, describing higher QoL. The mean satisfaction score from the whole group decreased between M0 and M18 (mainly in the youngests), and mainly concerned 3 fields (self-esteem, health and school) while their somatic health remained stable or improved. An effect of the familial context was also observed for these 3 fields. CONCLUSION: The progressive decline of QoL in HIV-infected children, and the surprising high and steady level of satisfaction over-time provided by the adolescents, underlined the frailty of this population and the need for a psychologic management associated to the medical follow-up. Such a multidisciplinary approach should take into account the preoccupations and difficulties of each age-class, those linked to the diagnosis itself, and to the familial or scolar contexts, in order to preserve QoL of this pediatric population, as far as possible, in a long term. PMID- 16364614 TI - [Magnetic resonance imaging features of desmoplastic cerebral ganglioglioma of infancy: report of 1 case]. AB - Desmoplastic infantile ganglioglioma is a rare intracranial tumor of infancy, characterized by solid and cystic component, voluminous size and supratentorial location. These tumors are diagnosed usually below the age of 2 years. We report 1 case of desmoplastic ganglioglioma in 13-year-old male. Computed tomography and magnetic resonance imaging diagnosed supratentorial mixed cystic and solid tumor, which presented as a large cystic component with intense contrast enhancement of a mural nodule. The tumor was surgically removed, and histology revealed desmoplastic ganglioglioma. The patient had a good follow up. This observation emphasizes the possibility of desmoplastic ganglioglioma in older infants. It mustn't be considered as a specific entity of very young age infant and must be recognized in older infant because it may be misdiagnosed as malignant glioma. Despite the pseudo malignant appearance, these tumors have a good prognosis after surgery and when excision is complete they don't led to recurrences. PMID- 16364615 TI - [Ifosfamide induced encephalopathy: 15 observations]. AB - INTRODUCTION: Ifosfamide is an alkylating agent used in the treatment of germ cell tumors, sarcomas and lymphomas. One of its main side effects is the encephalopathy of which the incidence may reach 30% in the literature, in adults and children just as well. OBJECTIVES: Based on both our experience and a review of the literature, we propose some recommendations for the management of this complication. PATIENTS AND METHODS: We report 15 encephalopathy cases in non brain tumor patients, which occurred between January 1987 and March 2002 in children from 2 to 17 years old, treated for solid tumors at the Institut Gustave Roussy. Ifosfamide was administered at a posology between 5.4 and 15 g/m(2)/course, associated with other antimitotics such as actinomycin D, etoposide or vincristine. RESULTS: Six patients experienced a grade III neurological toxicity according to the NCI classification, which developed as excess drowsiness lasting up to 36 hours. Six other patients developed grade IV neurotoxicity, including two comas resolving within 4 days and four short generalized convulsions. Three other children experienced grade II drowsiness. Brain MRIs were normal and EEG showed an aspecific encephalopathy tracing. This early central neurotoxicity appeared right from the first administration, and occurred immediately after the first injection or during the second or third day of treatment. It was most often reversible, usually 3 to 5 days after the last ifosfamide administration. Five patients were administered a treatment with Methylene Blue with a demonstrable efficacy in only one case. No death or neurological sequelae have been noted. Ifosfamide has been renewed after the neurological accident in 7 of those patients. Only 1 of those 7 patients developed grade IV neurotoxicity during the next course of treatment. In 2 of those 7 children, Methylene Blue was used in a prophylactic way. No neurological disorders have been noted during the next courses of treatment. DISCUSSION: In the literature, the following are described as risk factors for ifosfamide encephalopathy: advanced pelvic disease, previous cisplatyl treatment and renal failure. We have not found any of these predisposing factors in our series, but three of the fifteen patients had severe neurotoxicity associated with Vincristin during previous treatments. CONCLUSION: Facing a clinical diagnosis of ifosfamide encephalopathy, it is recommended to discontinue administration of ifosfamide and inject by intravenous route 50 mg Methylene Blue every 4 hours until the symptomatology recedes. The re-challenge of Ifosfamide is not contra-indicated and should be performed under prophylactic treatment with Methylene Blue by intravenous route at the dose of 50 mg every 6 hours. PMID- 16364616 TI - [Spontaneous rupture of a choledochal cyst in a 11-month-old girl]. AB - Choledochal cysts are rare congenital malformations of the biliary tract. Though most cysts are diagnosed incidentally, some present directly with complications. We report on the case of an 11-month-old girl admitted for abdominal pain, fever and vomiting. Ultrasonography revealed intraabdominal fluid and the absence of a choledochal cyst diagnosed 2 months earlier. Laparotomy for suspected rupture of a choledochal cyst was planned and a choledochojejunostomy with Roux-en-Y was performed. Spontaneous rupture of a choledochal cyst is rare and occurs most frequently in children under the age of 4. The exact cause is yet unknown and several factors have been implicated. The most probable cause is the combination of pancreatic reflux and epithelial irritation of a weakened cyst wall. Choledochal cysts should preferably be treated as soon as the child is 6 months old. Complete excision of the cyst is mandatory because of the risk of malignant transformation. PMID- 16364617 TI - Clinical decision-support for diagnosing stress-related disorders by applying psychophysiological medical knowledge to an instance-based learning system. AB - OBJECTIVE: An important procedure in diagnosing stress-related disorders caused by dysfunction in the interaction of the heart with breathing, i.e., respiratory sinus arrhythmia (RSA), is to analyse the breathing first and then the heart rate. Analysing these measurements is a time-consuming task for the diagnosing clinician. A decision-support system in this area would reduce the analysis task of the clinician and enable him/her to give more attention to the patient. We have created a decision-support system which contains a signal classifier and a pattern identifier. The system performs an analysis of the physiological time series concerned which would otherwise be performed manually by the clinician. METHODS: The signal-classifier, HR3Modul, classifies heart-rate patterns by analysing both cardio- and pulmonary signals, i.e., physiological time series. HR3Modul uses case-based reasoning (CBR), using a wavelet-based method for retrieving features from the signals. The system searches for familiar shapes in the signals by comparing them with shapes already stored. We have applied a best fit scheme for handling signals of different lengths, as the length of a breath is highly dynamic. We also apply automatic weighting to the features to obtain a more autonomous system. The classified heart signals indicate if a patient may be suffering from a stress-related disorder and the nature of the disorder. These classified signals are thereafter sent to the second subsystem, the pattern identifier. The pattern-identifier analyses the classified signals and searches for familiar patterns by identifying sequences in the classified signals. The identified sequences give clinicians a more complete analysis of the measurements, providing them with a better basis for diagnosis. RESULTS AND CONCLUSION: We have shown that a case-based classifier with a wavelet feature extractor and automatic weighting is a viable option for building a decision support system for the psychophysiological domain, as it is at par, or even outperforms other retrieval techniques and is less complex. PMID- 16364618 TI - On the role of methacrylic acid copolymers in the intracellular delivery of antisense oligonucleotides. AB - The delivery of active biomacromolecules to the cytoplasm is a major challenge as it is generally hindered by the endosomal/lysosomal barrier. Synthetic titratable polyanions can overcome this barrier by destabilizing membrane bilayers at pH values typically found in endosomes. This study investigates how anionic polyelectrolytes can enhance the cytoplasmic delivery of an antisense oligonucleotide (ODN). Novel methacrylic acid (MAA) copolymers were examined for their pH-sensitive properties and ability to destabilize cell membranes in a pH dependent manner. Ternary complex formulations prepared with the ODN, a cationic lipid and a MAA copolymer were systematically characterized with respect to their size, zeta potential, antisense activity, cytotoxicity and cellular uptake using the A549 human lung carcinoma cell line. The MAA copolymer substantially increased the activity of the antisense ODN in inhibiting the expression of protein kinase C-alpha. Uptake, cytotoxicity and antisense activity were strongly dependent on copolymer concentration. Metabolic inhibitors demonstrated that endocytosis was the major internalization pathway of the complexes, and that endosomal acidification was essential for ODN activity. Confocal microscopy analysis of cells incubated with fluorescently-labeled complexes revealed selective delivery of the ODN, but not of the copolymer, to the cytoplasm/nucleus. This study provides new insight into the mechanisms of intracellular delivery of macromolecular drugs, using synthetic anionic polyelectrolytes. PMID- 16364619 TI - n-6 and n-3 Long-chain polyunsaturated fatty acids in the erythrocyte membrane of Brazilian preterm and term neonates and their mothers at delivery. AB - Placental transfer of the long-chain polyunsaturated fatty acids (LCPUFA) arachidonic (AA) and docosahexaenoic (DHA) acids is selectively high to maintain accretion to fetal tissues, especially the brain. The objectives of the present study were to investigate the essential fatty acid (EFA) and LCPUFA status at birth of preterm and term Brazilian infants and their mothers, from a population of characteristically low intake of n-3 LCPUFA, and to evaluate the association between fetal and maternal status, by the determination of the fatty acid composition of the erythrocyte membrane. Blood samples from umbilical cord of preterm (26-36 weeks of gestation; n = 30) and term (37-42 weeks of gestation; n = 30) infants and the corresponding maternal venous blood were collected at delivery. The LCPUFA composition of the erythrocyte membrane and DHA status were similar for mothers of preterm and term infants. Neonatal AA was higher (P < 0.01) whereas its precursor 18:2n-6 was lower (P < 0.01) than maternal levels, as expected. There was no difference in LCPUFA erythrocyte composition between preterm and term infants, except for DHA. Term infants presented a worse DHA status than preterm infants (P < 0.01) and than their mothers (P < 0.01) at delivery. There was a negative correlation of neonatal DHA with maternal AA and a positive correlation between neonatal AA and maternal AA and 18:2n-6 only at term. These results suggest that the persistent low DHA maternal status, together with the comparatively better AA and 18:2n-6 status, might have affected maternal fetal transfer of DHA when gestation was completed up to term, and possibly contributed to the worse DHA status of term neonates compared with the preterm neonates. PMID- 16364620 TI - Aberrant expression of 5-lipoxygenase-activating protein (5-LOXAP) has prognostic and survival significance in patients with breast cancer. AB - 5-lipoxygenase (5-LOX)-activating protein, 5-LOXAP also known as LOX5AP or FLAP, is a protein that works closely with 5-LOX in regulating the metabolism of arachidonate. Some of the eicosanoid products of 5-LOX/5-LOXAP are known to play active roles in the function of cancer cells, including breast cancer cells. The current study investigated the expression of 5-LOXAP in clinical breast cancer and the prognostic impact of 5-LOXAP and 5-LOX in patients with breast cancer. A cohort of breast tumour tissues (n = 122) with normal background tissues (n = 32) were investigated. 5-LOXAP and 5-LOX transcripts were determined using RT-PCR and quantitative RT-PCR. Levels of the transcripts were analysed against clinical and pathological information. Breast tumour tissues had significantly higher levels of 5-LOX transcript compared with normal tissues (P = 0.015). The transcript was seen at significantly higher levels in node positive tumours than that in node negative tumours (P = 0.02). The prognostic significance was assessed using both a prognostic index and clinical outcome. Value of 5-LOXAP was first demonstrated when using the Nottingham Prognostic Index (NPI) as an indicator, in that patients with predicted poor prognosis had significantly higher levels of 5-LOXAP than patients with good prognosis (P = 0.0407). Furthermore, patients who died of breast cancer-related causes had significant higher levels of 5-LOXAP than those patients who remained disease free, following a median 10-year followup. A survival analysis has shown that high levels of 5-LOXAP were significantly correlated with overall survival (mean survival 109.6 month vs. 139.4 months, in tumour from patients with high and low levels of 5-LOXAP, P = 0.05). The same disadvantage of high levels of 5-LOXAP was also seen with disease-free survival (105.2 months vs. 135.6 months, P = 0.017). Analysis of 5-LOXAP together with 5 LOX transcript did not enhance the significance of the survival. However, when 5 LOXAP was considered together with 12-LOX, it improved the predictive power for both overall and disease-free survival (109.0 month vs. 143.1 months, P = 0.0156 for overall survival and 98.3 months vs. 141.3 months for disease-free survival, P = 0.0022). In conclusion, 5-LOXAP expression was aberrant in human breast cancer, particularly in aggressive tumours. Furthermore, 5-LOXAP had a significant prognostic value in patients with breast cancer. This identifies 5 LOXAP as a potential therapeutic target in breast cancer. PMID- 16364621 TI - Does Helicobacter pylori have a pathogenic role in bronchiectasis? AB - AIM: To investigate the presence of Helicobacter pylori (H. pylori) in bronchial biopsies of patients with bronchiectasis, by histochemical and immunochemical staining. SETTING: 800-bed tertiary university hospital. METHODS: Observational study. PATIENTS: forty-six patients with bronchiectasis in a stable clinical condition and 8 control patients. INTERVENTIONS: Serum samples determination of IgG levels for H. pylori by ELISA. Immunostaining with an anti-H. pylori antibody (NCL-HPp, Novocastra) of bronchial mucosa obtained by fiberoptic bronchoscopy from both patients with bronchiectasis and controls. RESULTS: Twenty-one out of 46 patients with bronchiectasis (46%) had positive serology for H. pylori. We obtained 40 bronchial biopsies in patients with bronchiectasis and 8 bronchial biopsies in control patients. No evidence of H. pylori was obtained in the bronchial samples of both patients and controls. CONCLUSIONS: The results of our study could not demonstrate the presence of H. pylori in bronchial specimens from patients with bronchiectasis. PMID- 16364622 TI - Asthma mortality, inhaled steroids, and changing asthma therapy in Argentina (1990-1999). AB - BACKGROUND: The asthma managing strategy has evolved and inhaled corticosteroids (ICS) are now the cornerstone of asthma treatment. Their effect on symptoms and exacerbations are well ascertained, whereas their effects on asthma mortality are still matter of debate. This ecological study evaluated asthma mortality rates and drug sales in the decade 1990-1999 in Argentina. METHODS: Mortality (overall and adjusted for 5-34 years) was obtained from the Argentinean Epidemiological Institute, and data on the sales of antiasthma drugs were provided by the International Marketing Survey. RESULTS: There was a significant change in drug sales in the spanned period: ICS +479%, inhaled beta2 agonists +32%, theophylline -63%. The crude and adjusted mortality rates were 3.38 and 0.72 in the 1980-1989 decade, and 2.58 (P<0.05) and 0.38 (P<0.01) in the subsequent one. There was inverse correlation between inhaled corticosteroid sale and age-adjusted (5-34) asthma mortality (r=-0.84; P=0.003), and the same with global mortality rates (r= 0.81; P=0.005). A positive correlation was also seen between theophylline sales and mortality. CONCLUSION: The increased sale of ICS and possibly the decrease of theophylline use seem to be the more relevant factors associated with decreased asthma mortality in Argentina. PMID- 16364623 TI - The nuclear envelope: form and reformation. AB - The membrane system that encloses genomic DNA is referred to as the nuclear envelope. However, with emerging roles in signaling and gene expression, these membranes clearly serve as more than just a physical barrier separating the nucleus and cytoplasm. Recent progress in our understanding of nuclear envelope architecture and composition has also revealed an intriguing connection between constituents of the nuclear envelope and human disease, providing further impetus to decipher this cellular structure and the dramatic remodeling process it undergoes with each cell division. PMID- 16364624 TI - Molecular details of formin-mediated actin assembly. AB - Formins are a large family of multi-domain polypeptides that form homodimers. The highly conserved formin homology 2 (FH2) domain and its neighboring formin homology 1 (FH1) domain, which are surrounded by regulatory domains, cooperate in rapidly assembling profilin-actin into long filaments while remaining continuously associated with the fast-growing barbed end. Recent biochemical, biophysical, theoretical and structural studies have concluded that diverse formins are mechanistically similar, but that the rates of various assembly states differ quantitatively, and have shed light on the mechanism of formin auto regulation and activation by Rho GTPases. PMID- 16364625 TI - PAR proteins and the cytoskeleton: a marriage of equals. AB - The PAR proteins are a group of widely conserved regulators of polarity, many of which are asymmetrically localized in polarized cells. Recent work shows that distinct modes of actomyosin- and microtubule-based transport contribute to the establishment of PAR asymmetries in different cell types. Cross-regulatory interactions among PAR proteins and with other conserved polarity complexes stabilize asymmetries once they form, and shape the evolution of PAR protein distributions in response to cytoskeletal transport or other polarizing inputs. The PAR proteins in turn modulate the actomyosin and microtubule cytoskeletons. In some cases, this is a form of feedback control, central to the establishment and maintenance of PAR asymmetries. In others, it underlies the elaboration of functional cell polarity. PMID- 16364626 TI - Small-scale medical waste incinerators--experiences and trials in South Africa. AB - Formal waste management services are not accessible for the majority of primary healthcare clinics on the African continent, and affordable and practicable technology solutions are required in the developing country context. In response, a protocol was established for the first quantitative and qualitative evaluation of relatively low cost small-scale incinerators for use at rural primary healthcare clinics. The protocol comprised the first phase of four, which defined the comprehensive trials of three incineration units. The trials showed that all of the units could be used to render medical waste non-infectious, and to destroy syringes or render needles unsuitable for reuse. Emission loads from the incinerators are higher than large-scale commercial incinerators, but a panel of experts considered the incinerators to be more acceptable compared to the other waste treatment and disposal options available in under-serviced rural areas. However, the incinerators must be used within a safe waste management programme that provides the necessary resources in the form of collection containers, maintenance support, acceptable energy sources, and understandable operational instructions for the incinerators, whilst minimising the exposure risks to emissions through the correct placement of the units in relation to the clinic and the surrounding communities. On-going training and awareness building are essential in order to ensure that the incinerators are correctly used as a sustainable waste treatment option. PMID- 16364627 TI - Effects of cement on redistribution of trace metals and dissolution of organics in sewage sludge and its inorganic waste-amended products. AB - The suitability of using cement-stabilized sludge products as artificial soils in earth works was evaluated. The sludge products investigated were cemented sludge, cement-treated clay-amended sludge (SS+MC), and cement-treated copper slag amended sludge (SS+CS). The leachability of lead (Pb), zinc (Zn), copper (Cu), and chromium (Cr) were assessed using the sequential extraction technique, toxicity characteristic leaching procedure (TCLP), NEN 7341 availability test, and column leaching test. The results indicated that Zn leachability was reduced in all the cement-stabilized sludge products. In contrast, Cu was transferred from the organic fraction to the readily leachable phases in the cement stabilized sludge products and therefore exhibited increased leachability. The increased Cu leachability could be attributed to dissolution of humic substances in the sludge as a result of elevated pH. Good correlation between dissolved organic carbon (DOC) and heavy metal leaching from the cement-stabilized sludge products was observed in the column leaching experiment. Even with a cement percentage as small as 12.5%, calcium silicate hydrate (C-S-H) was formed in the SS+MC and SS+CS products. Inclusion of the marine clay in the SS+MC products could reduce the leaching potentials of Zn, and this was the great advantage of the marine clay over the copper slag for sludge amendment. PMID- 16364628 TI - Release and trafficking of lipid-linked morphogens. AB - Wnt and Hedgehog family proteins are secreted morphogens that act on surrounding cells to pattern many different tissues in both vertebrates and invertebrates. The discovery that these proteins are covalently linked to lipids has raised the puzzling problem of how they come to be released from cells and move through tissue. A synergistic combination of biochemical, cell biological and genetic approaches over the past several years is beginning to illuminate both the forms in which lipid-linked morphogens are released from cells and the variety of molecular and cell biological mechanisms that control their dispersal. PMID- 16364629 TI - A comparison of viral RNA-dependent RNA polymerases. AB - Genome replication in picornaviruses is catalyzed by a virally encoded RNA dependent RNA polymerase, termed 3D. These viruses also use a small protein primer, named VPg, to initiate RNA replication. The recent explosion of structural information on picornaviral 3D polymerases has provided insights into the initiation of RNA synthesis and chain elongation. Comparing these data with results from previous structural analyses of viral RNA-dependent RNA polymerases that catalyze de novo RNA synthesis sheds light on the different strategies that these viruses use to initiate replication. PMID- 16364630 TI - A story with a good ending: tRNA 3'-end maturation by CCA-adding enzymes. AB - CCA-adding enzymes (tRNA nucleotidyltransferases) are responsible for the maturation or repair of the functional 3' end of tRNAs. These enzymes are remarkable because they polymerize the essential nucleotides CCA onto the 3' terminus of tRNA precursors without using a nucleic acid template. Recent crystal structures, plus three decades of enzymology, have revealed the elegant mechanisms by which CCA-adding enzymes achieve their substrate specificity in a nucleic acid template independent fashion. The class I CCA-adding enzyme employs both an arginine sidechain and backbone phosphates of the bound tRNA to recognize incoming nucleotides. It switches from C to A addition through changes in the size and shape of the nucleotide-binding pocket, which is progressively altered by the elongating 3' terminus of the tRNA. By contrast, the class II CCA-adding enzyme uses only amino acid sidechains, which form a protein template for incoming nucleotide selection. PMID- 16364631 TI - Prospective comparison of the impact on treatment decisions of whole-body magnetic resonance imaging and computed tomography in patients with metastatic malignant melanoma. AB - Patient management and treatment strategies for metastatic melanoma depend largely on the stage of metastatic disease. The aim of this study was to compare contrast-enhanced whole-body magnetic resonance imaging (wbMRI) and whole-body computed tomography (wbCT) to detect distant metastases for staging. A total of 43 patients (41 with completed wbCT and wbMRI examination) with known American Joint Committee on Cancer (AJCC) stage III-IV malignant melanoma were examined and 775 metastases were identified by both methods. Whole-body CT was able to detect 522 metastases, whereas wbMRI found 730 metastases. Whole-body CT identified 188 pulmonary metastases, compared with 143 metastases detected by wbMRI. In kidneys, adrenal glands and lymph nodes, respectively, wbCT and wbMRI detected the same number of lesions. Whole-body MRI detected more metastases than wbCT in liver (detection rate 122/199), spleen (26/54), subcutaneous tissue (39/61), muscle (4/11), bone marrow (23/132) and brain (15/25). Therapy was modified as a consequence of wbMRI findings in 10/41 (24%) patients. In conclusion, wbMRI detected clearly more malignant melanoma metastases in most organ systems with the exception of lung metastases. More accurate and complete staging by wbMRI has an impact on treatment strategy in about one-quarter of the patients. PMID- 16364632 TI - An economic evaluation of phosphorus recovery as struvite from digester supernatant. AB - Phosphorus can be recovered from wastewater through crystallisation of struvite, MgNH(4)PO(4).6H(2)O. Approximately 1 kg of struvite can be crystallised from 100 m(3) of wastewater. Crystallisation is profitable compared to chemical and biological removal of phosphorus due to savings from the reduction in (i) chemicals used for precipitation and sludge disposal; and (ii) downtime for cleaning unwanted struvite formed during chemical and biological removal. The struvite produced annually from a wastewater treatment plant that processed 100 m3/d, would be sufficient to apply on 2.6 ha of arable land, as fertilizer. If struvite were to be recovered from wastewater treatment plants worldwide, 0.63 million tons of phosphorus (as P(2)O(5)) could be harvested annually, reducing phosphate rock mining by 1.6%. Therefore, this technology could provide opportunities to recover phosphorus sustainably from waste streams and preserve phosphorus reserves. PMID- 16364633 TI - Select metal adsorption by activated carbon made from peanut shells. AB - Agricultural by-products, such as peanut shells, contribute large quantities of lignocellulosic waste to the environment each growing season; but few, if any, value-added uses exist for their disposal. The objective of this study was to convert peanut shells to activated carbons for use in adsorption of select metal ions, namely, cadmium (Cd2+), copper (Cu2+), lead (Pb2+), nickel (Ni2+) and zinc (Zn2+). Milled peanut shells were pyrolyzed in an inert atmosphere of nitrogen gas, and then activated with steam at different activation times. Following pyrolysis and activation, the carbons underwent air oxidation. The prepared carbons were evaluated either for adsorption efficiency or adsorption capacity; and these parameters were compared to the same parameters obtained from three commercial carbons, namely, DARCO 12x20, NORIT C GRAN and MINOTAUR. One of the peanut shell-based carbons had metal ion adsorption efficiencies greater than two of the three commercial carbons but somewhat less than but close to Minotaur. This study demonstrates that peanut shells can serve as a source for activated carbons with metal ion-removing potential and may serve as a replacement for coal based commercial carbons in applications that warrant their use. PMID- 16364634 TI - Composting rapidly reduces levels of extractable oxytetracycline in manure from therapeutically treated beef calves. AB - Oxytetracycline (OTC) is a broad-spectrum antibiotic used in livestock production. The widespread use and relative persistence of OTC may encourage development of antibiotic-resistant bacteria. The objective of this study was to determine whether composting would substantially reduce the concentration of OTC found in manure from medicated animals. The effect of OTC on composting was also investigated. Five beef calves were medicated for 5 days with 22 mg/kg/day of OTC. Approximately 23% of the OTC fed to the calves was recovered in the manure. Manure samples collected from calves prior to and after medication were mixed with straw and woodchips, and aliquots of the subsequent mixtures were treated in laboratory composters for 35 days. In addition, aliquots of the OTC-containing mixture were incubated at 25 degrees C or sterilized followed by incubation at 25 degrees C. The presence of OTC did not appear to affect composting processes. Within the first six days of composting, levels of extractable OTC in the compost mixture decreased from 115+/-8 microg/g dry weight to less than 6+/-1 microg/g dry weight (a 95% reduction). In contrast, levels of extractable OTC in room temperature incubated and sterilized mixtures decreased only 12-25% after 37 and 35 days, respectively. Levels of total heterotrophic bacteria and OTC-resistant bacteria in the finished compost mixture were roughly 30-fold higher and 10-fold lower, respectively, than levels in the mixture prior to composting. Although the basis of the OTC disappearance during composting is not known, the preponderence of OTC-sensitive bacteria and the decrease of OTC-resistant bacteria in the finished compost suggests that OTC residues have been rendered biologically inactive or unavailable. PMID- 16364635 TI - Biodegradation of bilge waste from Patagonia with an indigenous microbial community. AB - Oily residues that are generated in normal ship operation are considered hazardous wastes. A biodegradation assay with autochthonous microbiota of Bilge Waste Oily Phase (BWOP) was performed in a bioreactor under controlled conditions. Petroleum, diesel oil, and PAH degraders were isolated from bilge wastes. These bacteria belong to the genus Pseudomonas and are closely related to Pseudomonas stutzeri as shown by 16S rDNA phylogenetic analysis. The indigenous microbial community of the bilge waste was capable of biodegrading the BWOP (1% v/v) with biodegradation efficiencies of 70% for hexane extractable material (HEM), 68% for total hydrocarbons (TH) and 90% for total aromatics hydrocarbons (TA) in 14 days. Solid phase microextraction (SPME) was successfully applied to evaluate hydrocarbon evaporation in a control experiment and demonstrated a mass balance closure of 88%. The SPME and biodegradation results give useful information to improve and scale up the process for BWOP treatment. PMID- 16364636 TI - Optimization of lactose utilization in deproteinated whey by Kluyveromyces marxianus using response surface methodology (RSM). AB - Kluyveromyces marxianus Y-8281 yeast culture was utilized for the biological treatment of deproteinated whey wastewater in a batch system. Removal of lactose was optimized by the utilization of response surface methodology, RSM. The empirical model developed through RSM in terms of effective operational factors of medium pH, temperature, lactose and ammonia concentrations was found adequate to describe the treatment of deproteinated whey. Through the analysis, medium pH and temperature were found to be the most significant factors and an increment in both had a positive effect on lactose utilization, while lactose and ammonia concentrations had the least weight within the ranges investigated. Based on contour plots and variance analysis, optimum operational conditions for maximizing lactose removal were found to be 31 degrees C, 45 g/L whey powder concentration, 4 g/L total ammonium salt concentration and medium pH 6. Under the optimum operating conditions determined, 95% lactose removal was achieved after an 18-h fermentation. PMID- 16364637 TI - Stimulation on DNA triplet repeat strand slippage synthesis by the designed spirocycles. AB - The designed simpler chiral spirocyclic helical compounds that mimic the molecular architecture of the DNA bulge binder NCSi-gb have been prepared. It has been found that the synthesized spirocyclic compounds have strong stimulation effect on DNA slippage synthesis. Their stimulation activities on DNA strand slippage suggest that they may bind to or induce the formation of a non Watson Crick structure during in vitro replication of DNA triplet repeats. PMID- 16364638 TI - Aza-retinoids as novel retinoid X receptor-specific agonists. AB - A new structurally simple series of potent lipophilic aza-retinoids RXR agonists has been developed. SAR studies for the N-alkyl-azadienoic acids described here demonstrate that the RXR activity profile is sensitive to the N-alkyl chain length. Further, we have expanded the work to include azadienoic acids, which exhibited many accessible conformations leading to a better understanding of the SAR around the series. PMID- 16364639 TI - Optimization of a privileged structure leading to potent and selective human melanocortin subtype-4 receptor ligands. AB - Design and synthesis of potent MC4 selective agonists based on cyclohexylpiperidine derived cyclic urea, oxazolidinones, and sulfonamide based privileged structures are disclosed. PMID- 16364640 TI - 2-((1H-Azol-1-yl)methyl)-N-arylbenzamides: novel dual inhibitors of VEGFR-1/2 kinases. AB - Novel potent derivatives of (azol-1-yl)methyl-N-arylbenzamides with improved solubility (>3mM) are described as ATP-competitive inhibitors of vascular endothelial growth factor receptor 2 (VEGFR-2). Many compounds display VEGFR-2 inhibitory activity reaching IC(50)<100 nM in the enzymatic assay. The compounds also inhibit the related tyrosine kinase, VEGFR-1, with similar potencies. Several compounds containing bulky lipophilic substituents at the benzamide pharmacophore yielded 10- to 17-fold selectivity for the VEGFR-2 versus VEGFR-1 kinase. PMID- 16364641 TI - The existence of a second allosteric site on the M1 muscarinic acetylcholine receptor and its implications for drug design. AB - Fully flexible docking of KT5720, an allosteric modulator of the muscarinic receptors, was performed on a dynamic model of the M(1) muscarinic acetylcholine receptor. The results confirmed the existence of a second allosteric site, located on the intracellular face of the receptor. These results would be beneficial for the design of modulators of this receptor to be used as an effective alternative against the Alzheimer's disease. PMID- 16364642 TI - Synthesis and SAR of substituted tetrahydrocarbazole derivatives as new NPY-1 antagonists. AB - The SAR of a new series of tetrahydrocarbazole derivatives is described: the appropriate decoration of this template led to the identification of a new class of NPY-1 antagonists showing good in vitro potency and a promising in vivo pharmacokinetic profile in rat. PMID- 16364643 TI - (+)-12alpha-Hydroxysophocarpine, a new quinolizidine alkaloid and related anti HBV alkaloids from Sophora flavescens. AB - (+)-12alpha-Hydroxysophocarpine (8), a new quinolizidine alkaloid was isolated from the roots of Sophora flavescens, together with 10 known quinolizidine alkaloids, (+)-oxymatrine (1), (+)-matrine (2), (+)-9alpha-hydroxymatrine (3), (+)-allomatrine (4), (+)-oxysophocarpine (5), (-)-sophocarpine (6), (-)-9alpha hydroxysophocarpine (7), (+)-lehmannine (9), (-)-13,14-dehydrosophoridine (10), and (-)-anagyrine (11). Their structures were elucidated by spectroscopic methods, and the stereochemistry of 8 was confirmed by X-ray analysis. These alkaloids were tested for anti-hepatitis B virus (HBV) activity in vitro, compounds 5, 6, 9, and 10 showed significant anti-HBV activity with inhibitory potency against HBsAg secretion at 48.3-79.3% and that against HBeAg secretion at 24.6-34.6%. PMID- 16364645 TI - Management of chylous leakage after axillary lymph node dissection in a patient undergoing breast surgery. AB - A 71-year old woman who underwent a modified radical mastectomy for invasive ductal carcinoma of the left breast, developed postoperative chylous leakage. Though conservative management was uneffective, a direct surgical repair led to good results. Because the morbidity of a reoperation to the superficial chest wall is low, timely surgical treatment is therefore strongly recommended in cases of high output chylous leakage following a mastectomy. PMID- 16364646 TI - Breast reduction and subsequent cancer: a prophylactic perspective. AB - The aim of this paper was to evaluate the impact of breast-gland remodelling, for cosmetic or functional purposes, on cancer incidence during long-term post surgical follow-up. We reviewed the literature investigating the ratio between the amount of breast tissue resected and cancer incidence during follow-up. Our analysis of the published data suggested that hypertrophic breast remodelling decreases the risk of breast and other types of cancer in post-operative patients. The actual risk reduction for patients over 40 years of age is related to the weight of the surgical specimens during the previous operation. Our conclusions support the use of breast-reduction surgery as a preventive measure in patients complaining of symptomatic breast enlargement, especially those with a family history of breast cancer. PMID- 16364647 TI - Phosphatidylinositol 3,5-bisphosphate: metabolism and cellular functions. AB - Polyphosphoinositides (PPIn) are low-abundance membrane phospholipids that each bind to a distinctive set of effector proteins and, thereby, regulate a characteristic suite of cellular processes. Major functions of phosphatidylinositol 3,5-bisphosphate [PtdIns(3,5)P(2)] are in membrane and protein trafficking, and in pH control in the endosome-lysosome axis. Recently identified PtdIns(3,5)P(2) effectors include a family of novel beta-propeller proteins, for which we propose the name PROPPINs [for beta-propeller(s) that binds PPIn], and possibly proteins of the epsin and CHMP (charged multi-vesicular body proteins) families. All eukaryotes, with the exception of some pathogenic protists and microsporidians, possess proteins needed for the formation, metabolism and functions of PtdIns(3,5)P(2). The importance of PtdIns(3,5)P(2) for normal cell function is underscored by recent evidence for its involvement in mammalian cell responses to insulin and for PtdIns(3,5)P(2) dysfunction in the human genetic conditions X-linked myotubular myopathy, Type-4B Charcot-Marie Tooth disease and fleck corneal dystrophy. PMID- 16364648 TI - Quantitative chemical analysis of ocular melanosomes in the TEM. AB - Melanosomes in retinal tissues of a human, monkey and rat were analyzed by EDX in the TEM. Samples were prepared by ultramicrotomy at different thicknesses. The material was mounted on Al grids and samples were analyzed in a Zeiss 912 TEM equipped with an Omega filter and EDX detector with ultrathin window. Melanosomes consist of C and O as main components, mole fractions are about 90 and 3-10 at.%, respectively, and small mole fraction ratios, between 2 and 0.1 at.%, of Na, Mg, K, Si, P, S, Cl, Ca. All elements were measured quantitatively by standardless EDX with high precision. Mole fractions of transition metals Fe, Cu and Zn were also measured. For Fe a mole fraction ratio of less than 0.1at.% was found and gives the melanin its paramagnetic properties. Its mole fraction is however close to or below the minimum detectable mass fraction of the used equipment. Only in the human eye and only in the retinal pigment epitelium (rpe) the mole fractions of Zn (0.1 at.% or 5000 microg/g) and Cu were clearly beyond the minimum detectable mass fraction. In the rat and monkey eye the mole fraction of Zn was at or below the minimum detectable mass fraction and could not be measured quantitatively. The obtained results yielded the chemical composition of the melanosomes in the choroidal tissue and the retinal pigment epitelium (rpe) of the three different species. The results of the chemical analysis are discussed by mole fraction correlation diagrams. Similarities and differences between the different species are outlined. Correlation behavior was found to hold over species, e.g. the Ca-O correlation. It indicates that Ca is bound to oxygen rich sites in the melanin. These are the first quantitative analyses of melanosomes by EDX reported so far. The quantitative chemical analysis should open a deeper understanding of the metabolic processes in the eye that are of central importance for the understanding of a large number of eye-related diseases. The chemical analysis also allows a correlation with structural changes observed at the various regions of the eye. PMID- 16364649 TI - Phagocytosis, endosomal/lysosomal system and other cellularaspects of macrophage activation by Canova medication. AB - Canova is a homeopathic medication with immunomodulatory properties, recommended for diseases where the immune system is depressed. Our research aims to study the activation of mice peritoneal macrophages when submitted to in vivo and in vitro Canova treatment. Morphological parameters and acid phosphatase activity were analyzed using light and transmission electron microscopy. Differential interference contrast microscopy, including serial time acquisition in living cells, was also performed. The results demonstrated a greater spreading ability in Canova treated macrophages, a higher phagocytic activity of non-infective microorganisms (Saccharomyces cerevisiae and Tripanosoma cruzi epimastigotes) and a tendency to lower the phagocytic activity of the infective microorganisms T. cruzi trypomastigotes and Leishmania amazonensis, when compared with control cells. Acid phosphatase activity was analyzed and showed that Canova treatment stimulates an increase of the endosomal/lysosomal system. Treated macrophages that do or do not interact with yeast present a higher number of acid phosphatase marked vesicles compared to control cells. In contrast, the activity of tartrate resistant acid phosphatase (TRAP), is lower in Canova treated macrophages. The net results demonstrate that Canova medication is an effective stimulator of macrophage activity. PMID- 16364650 TI - Induction of apoptosis in cells expressing exogenous Hippi, a molecular partner of huntingtin-interacting protein Hip1. AB - To decipher the pathway of apoptosis induction downstream to caspase-8 activation by exogenous expression of Hippi, an interactor of huntingtin-interacting protein Hip1, we studied apoptosis in HeLa and Neuro2A cells expressing GFP-tagged Hippi. Nuclear fragmentation, caspase-1, caspase-8, caspase-9/caspase-6 and caspase-3 activation were increased significantly in Hippi expressing cells. Cleavage of Bid, release of cytochrome c and apoptosis inducing factor (AIF) from mitochondria were also increased in GFP-Hippi expressing cells. It was observed that caspase-1 and caspase-8 activation was earlier than caspase-3 activation and nuclear fragmentation. Expression of caspase-1, caspase-3 and caspase-7 was increased while anti-apoptotic gene Bcl-2 and mitochondrial genes ND1 and ND4 were reduced in Hippi expressing cells. Besides, the expression SDHA and SDHB, nuclear genes, subunits of mitochondrial complex II were decreased in GFP-Hippi expressing cells. Taken together, we concluded that Hippi expression induced apoptosis by releasing AIF and cytochrome c from mitochondria, activation of caspase-1 and caspase-3, and altering the expression of apoptotic genes and genes involved in mitochondrial complex I and II. PMID- 16364651 TI - The endocannabinoid 2-AG protects the blood-brain barrier after closed head injury and inhibits mRNA expression of proinflammatory cytokines. AB - Endocannabinoids are involved in neuroprotection through numerous biochemical pathways. We have shown that the endocannabinoid 2-arachidonoyl glycerol (2-AG) is released in mouse brain after closed head injury (CHI), and treatment with exogenous 2-AG exerts neuroprotection via the central cannabinoid receptor CB1. This process involves inhibition of inflammatory signals that are mediated by activation of the transcription factor NF-kB. The present study was designed to examine the effect of 2-AG on the blood-brain barrier (BBB) and the possible inhibition of the early expression of proinflammatory cytokines, which are implicated in BBB disruption. We found that 2-AG decreased BBB permeability and inhibited the acute expression of the main proinflammatory cytokines: TNF-alpha, IL-1beta and IL-6. It also augmented the levels of endogenous antioxidants. We suggest that 2-AG exerts neuroprotection in part by inhibition of the early (1-4 h) inflammatory response and augmentation of the brain reducing power. PMID- 16364652 TI - Discovery of histamine H3 receptor antagonistic property of simple imidazole-free derivatives: Preliminary pharmacological investigation. AB - The histamine H3 receptor subtype negatively modulates the release of various neurotransmitters such as histamine, glutamate, norepinephrine, acetylcholine and many others mainly in the CNS and H3 antagonists have been developed to treat central diseases characterized by neurotransmission disturbance such as schizophrenia, memory/learning and sleep disorders. In search for non-imidazole histamine H3 receptor antagonists, currently indicated as a promising class of H3 blockers, a series of simple alkylpiperidine derivatives has been studied to attain a preliminary pharmacological profile. The compounds were characterized in vitro in terms of binding affinity, antagonistic potency and selectivity at rodent H3 receptors. The imidazole-free derivatives possessed moderate to pronounced antagonistic potency at guinea-pig ileal H3 receptor consistent with binding affinity at rat brain H3 receptors and showed a favourable receptor selectivity profile. For the compound 5, with the highest affinity at rat H3 receptors, comparable values were calculated in binding (pKi = 8.35) and functional (pA2 = 8.22) assays in SK-N-MC cells stably expressing human H3 receptors. These findings indicate to extend the investigation to pharmacokinetic property and central effects to gain deeper knowledge on the pharmacological potential of this compound. PMID- 16364653 TI - Clustering transmembrane-agrin induces filopodia-like processes on axons and dendrites. AB - The transmembrane form of agrin (TM-agrin) is primarily expressed in the CNS, particularly on neurites. To analyze its function, we clustered TM-agrin on neurons using anti-agrin antibodies. On axons from the chick CNS and PNS as well as on axons and dendrites from mouse hippocampal neurons anti-agrin antibodies induced the dose- and time-dependent formation of numerous filopodia-like processes. The processes appeared within minutes after antibody addition and contained a complex cytoskeleton. Formation of processes required calcium, could be inhibited by cytochalasine D, but was not influenced by staurosporine, heparin or pervanadate. Time-lapse video microscopy revealed that the processes were dynamic and extended laterally along the entire length of the neuron. The lateral processes had growth cones at their tips that initially adhered to the substrate, but subsequently collapsed and were retracted. These data provide the first evidence for a specific role of TM-agrin in shaping the cytoskeleton of neurites in the developing nervous system. PMID- 16364654 TI - The final fates of neurogenin2-expressing cells include all major neuron types in the mouse retina. AB - The molecular mechanism underlying vertebrate retinal development is not well understood. To examine whether neurogenin2 (ngn2) expression determines cell fate in the retina, we mapped the final fates of cells that once expressed ngn2, using the conditional, binary CreER -LacZ system. We found LacZ+ cells in all 3 nuclear layers of the mouse retina and including all major types of neurons: photoreceptors, horizontal, bipolar, amacrine, and ganglion cells. The distribution of LacZ+ cells among the 3 nuclear layers closely resembled a theoretical distribution of total retinal cells. The temporal window in which each cell type was marked appeared nonrandom, and was similar to its birthdate and birth sequence. These data indicate that cells expressing ngn2 at some point in their life histories may later differentiate into a variety of cell types. PMID- 16364655 TI - Gi/o proteins: expression for direct activation enquiry. AB - G protein-mediated pathways are fundamental mechanisms of cell signaling. In this paper, the expression and the characterization of the alphai1, alphai3, alphao1, beta1, and gamma2 subunits of the human G protein are described. This approach was developed to evaluate the G protein activation profile of new compounds. pCR TOPO T7 vectors, engineered to contain the target sequences, were used to transform Escherichia coli competent cells. Subunits were over-expressed in a preparative scale as fusion proteins with a six-histidine tag, and subsequently purified by metal chelate chromatography. Afterward, the His-tag was removed by enterokinase digestion, and the secondary structures of the recombinant subunits were analyzed by circular dichroism. To assess the functionality of the subunits, the rate of GTP hydrolysis and GTPgammaS binding were evaluated both in the absence and in the presence of two modulators: the peptidic activator Mastoparan and the non-peptidic activator N-dodecyl-lysinamide (ML250). Tests were conducted on isolated alpha-subunit and on heterotrimeric alphabetagamma complex, alone or reconstituted in phospholipidic vesicles. Our results show that recombinant subunits are stable, properly folded and, fully active, which makes them suitable candidates for functional studies. PMID- 16364656 TI - Cloning and expression of superoxide dismutase from Mycobacterium bovis BCG. AB - We have previously purified the superoxide dismutase (SOD) of Mycobacterium bovis bacillus Calmette-Guerin (BCG), and there is no signal peptide necessary for protein exportation [S.K. Kang, Y.J. Jung, C.H. Kim, C.Y. Song, Extracellular and cytosolic iron superoxide dismutase from Mycobacterium bovis BCG, Clin. Diagn. Lab. Immunol. 5 (1998) 784-789]. In the present study, SOD gene of M. bovis BCG was cloned and expressed in Escherichia coli, and its complete nucleotide sequence and deduced amino acid composition were determined. The open reading frame from the GTG initiation codon was 621 base pair (bp) in length for the SOD structural gene. The ribosomal-binding sequences (GGAAGG) were 6-12 bp upstream from the initiation codon. The amino acid sequence, deduced from the nucleotide sequence, revealed that the SOD consists of 207 amino acids residues with a molecular weight of 22.8 kDa. The N-terminal amino acid sequence predicted from the nucleotide sequence showed that the structural gene of the SOD is not preceded by leader sequences. There were no cysteine residues in the deduced amino acid composition, indicating that the SOD does not consist of disulfide bonds. Analyses of both nucleotide and amino acid sequences of the SOD showed significant similarity to other pathogenic mycobacterial SODs. Furthermore, the results of fractionation and two-dimensional electrophoresis showed that SOD is also associated with cell membrane, suggesting that there might be a specific mechanism for exportation of SOD in M. bovis BCG as well as other pathogenic mycobacteria. Overexpressed SOD in E. coli was purified from the inclusion bodies, and the histidine tag was removed from the protein using enterokinase. Enzyme activity was then determined by gel staining analysis. PMID- 16364657 TI - Study of the localization of iron, ferritin, and hemosiderin in Alzheimer's disease hippocampus by analytical microscopy at the subcellular level. AB - Previous studies of the structure of core nanocrystals of ferritin (Ft) in the brains of patients with Alzheimer's disease (AD) have shown differences in the mineral compound in comparison with physiological Ft. Both Ft cores have a polyphasic composition but whereas the major phase in physiological Ft is hexagonal ferric iron oxide (ferrihydrite), the major phases in brain AD Ft are two cubic mixed ferric-ferrous iron oxides (magnetite and wustite). One of these (wustite) is similar to what is detected in hemosiderin (Hm) cores in primary hemochromatosis (Quintana, C., Cowley, J.M, Marhic, C., 2004. Electron nanodiffraction and high resolution electron microscopy studies of the structure and composition of physiological and pathological ferritin. J. Struct. Biol. 147, 166-178). We have studied, herein, the distribution of iron, Ft, and Hm in sections of AD hippocampus using analytical microscopy. Iron present in Ft cores was directly mapped in a nanoSIMS microscope and the iron distribution has been correlated with the constituent elements N, P, and S. Ft and Hm cores were visualized at an ultrastructural level in an analytical transmission electron microscope. In senile plaques, Ft was observed in the coronal region associated with a non-beta-amyloid component and in the periphery of plaques, together with Hm, in sulfur-rich dense bodies of dystrophic neurites. Hm was also found in lysosomes and siderosomes of glial cells. Ft was observed in the cytoplasm and nucleus of oligodendrocytes. Ft was particularly abundant in myelinated axons in association with oligodendrocyte processes. These findings provide new arguments to support the hypothesis of a dysfunction of Ft (with eventual degradation to Hm) in AD resulting in an increase of toxic brain ferrous ions that may contribute to the production of free radicals that induce both cellular oxidative stress and aged-related myelin breakdown associated with cognitive decline and AD (Bartzokis, G., 2004. Age-related myelin breakdown: a developmental model of cognitive decline and Alzheimer's disease. Neurobiol. Aging 25, 5-18). PMID- 16364658 TI - Magnetic resonance imaging of the siliceous skeleton of the demosponge Lubomirskia baicalensis. AB - The skeletal elements (spicules) of the demosponge Lubomirskia baicalensis were analyzed; they are composed of amorphous, non-crystalline silica, and contain in a central axial canal the axial filament which consists of the enzyme silicatein. The axial filament, that orients the spicule in its longitudinal axis exists also in the center of the spines which decorate the spicule. During growth of the sponge, new serially arranged modules which are formed from longitudinally arranged spicule bundles are added at the tip of the branches. X-ray analysis revealed that these serial modules are separated from each other by septate zones (annuli). We describe that the longitudinal bundles of spicules of a new module originate from the apex of the earlier module from where they protrude. A cross section through the oscular/apical-basal axis shows that the bundle rays are organized in a concentric and radiate pattern. High resolution magnetic resonance microimaging studies showed that the silica spheres of the spicules in the cone region contain high amounts of 'mobile' water. We conclude that the radiate accretive growth pattern of sponges is initiated in the apical region (cones) by newly growing spicules which are characterized by high amounts of 'mobile' water; subsequently spicule bundles are formed laterally around the cones. PMID- 16364659 TI - Intraoperative visualization of the pyramidal tract by diffusion-tensor-imaging based fiber tracking. AB - Functional neuronavigation allows intraoperative visualization of cortical eloquent brain areas. Major white matter tracts, such as the pyramidal tract, can be delineated by diffusion-tensor-imaging based fiber tracking. These tractography data were integrated into 3-D datasets applied for neuronavigation by rigid registration of the diffusion images with standard anatomical image data so that their course could be superimposed onto the surgical field during resection of gliomas. Intraoperative high-field magnetic resonance imaging was used to compensate for the effects of brain shift, which amounted up to 8 mm. Despite image distortion of echo planar images, which was identified by non linear registration techniques, navigation was reliable. In none of the 19 patients new postoperative neurological deficits were encountered. Intraoperative visualization of major white matter tracts allows save resection of gliomas near eloquent brain areas. A possible shifting of the pyramidal tract has to be taken into account after major tumor parts are resected. PMID- 16364660 TI - Gamma-band activity over early sensory areas predicts detection of changes in audiovisual speech stimuli. AB - Oscillatory activity in the gamma-band range in human magneto- and electroencephalogram is thought to reflect the oscillatory synchronization of cortical networks. Findings of enhanced gamma-band activity (GBA) during cognitive processes like gestalt perception, attention and memory have led to the notion that GBA may reflect the activation of internal object representations. However, there is little direct evidence suggesting that GBA is related to subjective perceptual experience. In the present study, magnetoencephalogram was recorded during an audiovisual oddball paradigm with infrequent visual (auditory /ta/ + visual /pa/) or acoustic deviants (auditory /pa/ + visual /ta/) interspersed in a sequence of frequent audiovisual standard stimuli (auditory /ta/ + visual /ta/). Sixteen human subjects had to respond to perceived acoustic changes which could be produced either by real acoustic or illusory (visual) deviants. Statistical probability mapping served to identify correlations between oscillatory activity in response to visual and acoustic deviants, respectively, and the detection rates for either type of deviant. The perception of illusory acoustic changes induced by visual deviants was closely associated with gamma band amplitude at approximately 80 Hz between 250 and 350 ms over midline occipital cortex. In contrast, the detection of real acoustic deviants correlated positively with induced GBA at approximately 42 Hz between 200 and 300 ms over left superior temporal cortex and negatively with evoked gamma responses at approximately 41 Hz between 220 and 240 ms over occipital areas. These findings support the relevance of high-frequency oscillatory activity over early sensory areas for perceptual experience. PMID- 16364661 TI - A Bayesian approach to modeling dynamic effective connectivity with fMRI data. AB - A state-space modeling approach for examining dynamic relationship between multiple brain regions was proposed in Ho, Ombao and Shumway (Ho, M.R., Ombao, H., Shumway, R., 2005. A State-Space Approach to Modelling Brain Dynamics to Appear in Statistica Sinica). Their approach assumed that the quantity representing the influence of one neuronal system over another, or effective connectivity, is time-invariant. However, more and more empirical evidence suggests that the connectivity between brain areas may be dynamic which calls for temporal modeling of effective connectivity. A Bayesian approach is proposed to solve this problem in this paper. Our approach first decomposes the observed time series into measurement error and the BOLD (blood oxygenation level-dependent) signals. To capture the complexities of the dynamic processes in the brain, region-specific activations are subsequently modeled, as a linear function of the BOLD signals history at other brain regions. The coefficients in these linear functions represent effective connectivity between the regions under consideration. They are further assumed to follow a random walk process so to characterize the dynamic nature of brain connectivity. We also consider the temporal dependence that may be present in the measurement errors. ML-II method (Berger, J.O., 1985. Statistical Decision Theory and Bayesian Analysis (2nd ed.). Springer, New York) was employed to estimate the hyperparameters in the model and Bayes factor was used to compare among competing models. Statistical inference of the effective connectivity coefficients was based on their posterior distributions and the corresponding Bayesian credible regions (Carlin, B.P., Louis, T.A., 2000. Bayes and Empirical Bayes Methods for Data Analysis (2nd ed.). Chapman and Hall, Boca Raton). The proposed method was applied to a functional magnetic resonance imaging data set and results support the theory of attentional control network and demonstrate that this network is dynamic in nature. PMID- 16364662 TI - Influence of tissue conductivity anisotropy on EEG/MEG field and return current computation in a realistic head model: a simulation and visualization study using high-resolution finite element modeling. AB - To achieve a deeper understanding of the brain, scientists, and clinicians use electroencephalography (EEG) and magnetoencephalography (MEG) inverse methods to reconstruct sources in the cortical sheet of the human brain. The influence of structural and electrical anisotropy in both the skull and the white matter on the EEG and MEG source reconstruction is not well understood. In this paper, we report on a study of the sensitivity to tissue anisotropy of the EEG/MEG forward problem for deep and superficial neocortical sources with differing orientation components in an anatomically accurate model of the human head. The goal of the study was to gain insight into the effect of anisotropy of skull and white matter conductivity through the visualization of field distributions, isopotential surfaces, and return current flow and through statistical error measures. One implicit premise of the study is that factors that affect the accuracy of the forward solution will have at least as strong an influence over solutions to the associated inverse problem. Major findings of the study include (1) anisotropic white matter conductivity causes return currents to flow in directions parallel to the white matter fiber tracts; (2) skull anisotropy has a smearing effect on the forward potential computation; and (3) the deeper a source lies and the more it is surrounded by anisotropic tissue, the larger the influence of this anisotropy on the resulting electric and magnetic fields. Therefore, for the EEG, the presence of tissue anisotropy both for the skull and white matter compartment substantially compromises the forward potential computation and as a consequence, the inverse source reconstruction. In contrast, for the MEG, only the anisotropy of the white matter compartment has a significant effect. Finally, return currents with high amplitudes were found in the highly conducting cerebrospinal fluid compartment, underscoring the need for accurate modeling of this space. PMID- 16364663 TI - New insights into the phylogeny of fig pollinators using Bayesian analyses. AB - The interaction between figs and fig pollinators is one of the most species specific mutualisms. Recently, phylogenies of both partners based on molecular data provided insights into a wide spectrum of co-evolutionary questions. However, for the phylogeny of fig pollinators, there are some discrepancies between different studies and left some relationships unresolved, especially for deep nodes. The phylogenetic uncertainties of pollinators prohibit our further understanding of the history of the mutualism. Here, we present phylogenetic analyses of a larger COI sequence dataset that includes previously published datasets and our sequences from 20 species using Bayesian method and maximum parsimony. The analyses using different methods share similar topologies. Bayesian analyses provide high level of confidence for most internal nodes in terms of posterior probability. This study also clarifies some discrepancies between previous studies. After rooting with Tetrapus, other pollinators split into two clades. Wiebesia and Blastophaga are at basal positions in respective clade. Ceratosolen is not monophyletic because Kradibia and Liporrhopalum fall inside this group. Three subgenera of Ceratosolen: subgen. Ceratosolen, subgen. Rothropus, and subgen. Strepitus are not supported. Therefore, Ceratosolen is suggested to be re-divided into three groups. Urostigma pollinators (including Dolichoris and Blastophaga psenes) are clustered together. The monophylies of Wiebesia, Blastophaga, Dolichoris are not supported in this analysis. This study also provides a new framework for re-evaluating character evolution and re inspecting the definition of some genera. PMID- 16364664 TI - Comparing alignment methods for inferring the history of the new world lizard genus Mabuya (Squamata: Scincidae). AB - The rapid increase in the ability to generate molecular data, and the focus on model-based methods for tree reconstruction have greatly advanced the use of phylogenetics in many fields. The recent flurry of new analytical techniques has focused almost solely on tree reconstruction, whereas alignment issues have received far less attention. In this paper, we use a diverse sampling of gene regions from lizards of the genus Mabuya to compare the impact, on phylogeny estimation, of new maximum likelihood alignment algorithms with more widely used methods. Sequences aligned under different optimality criteria are analyzed using partitioned Bayesian analysis with independent models and parameter settings for each gene region, and the most strongly supported phylogenetic hypothesis is then used to test the hypothesis of two colonizations of the New World by African scincid lizards. Our results show that the consistent use of model-based methods in both alignment and tree reconstruction leads to trees with more optimal likelihood scores than the use of independent criteria in alignment and tree reconstruction. We corroborate and extend earlier evidence for two independent colonizations of South America by scincid lizards. Relationships within South American Mabuya are found to be in need of taxonomic revision, specifically complexes under the names M. heathi, M. agilis, and M. bistriata (sensu, M.T. Rodrigues, Papeis Avulsos de Zoologia 41 (2000) 313). PMID- 16364665 TI - Uplift of the Tibetan plateau: evidence from divergence times of glyptosternoid catfishes. PMID- 16364666 TI - Intraoperative monitoring of penile and buttock blood flow during endovascular abdominal aortic aneurysm repair. AB - OBJECTIVE: The purpose of this study was to assess the pelvic circulation during endovascular abdominal aortic aneurysm repair (EVAR) with a new monitoring system measuring penile and buttock blood flow. METHODS: We measured penile brachial pressure index (PBI) during EVAR by pulse-volume-plethysmography (form PWV/ABItrade mark). We also measured bilateral gluteal tissue oxygen metabolism with near-infrared spectroscopy to provide a gluteal tissue oxygenation index (TOI). Twenty-two men who underwent aortouni-iliac stentgraft with crossover bypass for exclusion of abdominal aortic aneurysm were studied. Twelve patients underwent aorto-uni-common iliac artery stentgraft (CIA) and ten underwent aorto uni-external iliac artery stentgraft (EIA). RESULTS: In all patients, there was an immediate reduction in PBI during the EVAR procedure. After revascularization of the ipsilateral limb of the stent graft, the recovery of PBI was significantly less in EIA group. After the completion of crossover bypass, PBI in both groups recovered to the baseline values. In both groups there was a bilateral reduction in gluteal TOI during malperfusion of the internal iliac artery. After revascularization of ipsilateral limb of the stent graft, the ipsilateral TOI recovered to the baseline level in CIA patients, but recovery was incomplete in EIA patients. In contrast, contra-lateral TOI remained low in both groups after revascularization of ipsilateral limb of the stent graft. Only after completion of crossover bypass did the contra-lateral TOI recover to baseline level in both groups. CONCLUSIONS: Both TOI at the buttocks and PBI are a sensitive reflection of pelvic haemodynamics. Penile blood flow and bilateral gluteal blood flow are supplied via different circulations and both should be monitored for full assessment of the pelvic circulation. PMID- 16364668 TI - Spectral unraveling by space-selective Hadamard spectroscopy. AB - Spectral unraveling by space-selective Hadamard spectroscopy (SUSHY) enables recording of NMR spectra of multiple samples loaded in multiple sample tubes in a modified spinner turbine and a regular 5mm liquids NMR probe equipped with a tri axis pulsed field gradient coil. The individual spectrum from each sample is extracted by adding and subtracting data that are simultaneously obtained from all the tubes based on the principles of spatially resolved Hadamard spectroscopy. The well-known Hadamard spectroscopy has been applied for spatial selection and the method utilizes standard configuration of NMR instrument hardware. The SUSHY method can be easily incorporated in multi-dimensional multi tube NMR experiments. This method combines the excitation multiplexing, natural advantage of FTNMR, and sample multiplexing and offers high-throughput by reducing the total experimental time by up to a factor of four in a 4-tube mode. PMID- 16364667 TI - Mechanisms involved in the swelling of erythrocytes caused by Pacific and Caribbean ciguatoxins. AB - The mechanisms underlying the swelling of frog red blood cells (RBC), induced by Pacific (P-CTX-1) and Caribbean (C-CTX-1) ciguatoxins (CTXs), were investigated by measuring the length, width and surface of their elliptic shape. P-CTX-1 (0.5 to 5 nM) and C-CTX-1 (1 nM) induced RBC swelling within 60 min. The CTXs-induced RBC swelling was blocked by apamin (1 microM) and by Sr(2+) (1 mM). P-CTX-1 induced RBC swelling was prevented and inhibited by H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (27 microM), an inhibitor of soluble guanylate cyclase (sGC), and NOS blockade by NG methyl-l-arginine (l-NMA; 10 microM). Cytochalasin D (cytD, 10 microM) increased RBC surface and mimicked CTX effect but did not prevent the P-CTX-1-induced l-NMA-sensitive extra increase. Calculations revealed that P-CTX-1 and cytD increase RBC total surface envelop and volume. These data strongly suggest that the molecular mechanisms underlying CTXs-induced RBC swelling involve the NO pathway by an activation of the inducible NOS, leading to sGC activation which modulates intracellular cGMP and regulates L-type Ca(2+) channels. The resulting increase in intracellular Ca(2+) content, in turn, disrupts the actin cytoskeleton, which causes a water influx and triggers a Ca(2+)-activated K(+) current through SK2 isoform channels. PMID- 16364669 TI - The triple neurokinin-receptor antagonist CS-003 inhibits neurokinin A-induced bronchoconstriction in patients with asthma. AB - Neurokinin A (NKA) causes bronchoconstriction in asthmatic patients. In vitro both NK1 and NK2 receptors can mediate airway contraction. Moreover in guinea pigs, NK3 receptors facilitate cholinergic neurotransmission. Dual tachykinin NK1/NK2 receptor antagonism results in prevention of NKA-induced bronchoconstriction. We have now examined the effect of a single dose of the triple tachykinin receptor antagonist CS-003 on NKA-induced bronchoconstriction in asthmatics. A double blind, crossover, placebo-controlled trial in 16 mild asthmatics was performed. One single dose of CS-003 (200 mg, solution in distilled water) or matched placebo was given orally on the assessment days. NKA provocation tests were performed pre-dose and 1, 8 and 24 h after dosing. There was a significant shift to the right of the dose-response curve at 1 and 8 h after intake of CS-003. PC20 was not reached in 12/16 patients at 1h post-dose and in 5/16 patients at 8 h post-dose. This did not occur under placebo treatment. A single dose of 200 mg CS-003 protected significantly against NKA induced bronchoconstriction at 1 and 8h post-dose in mild asthmatics. PMID- 16364670 TI - Chilling-injury and disturbance of ion homeostasis in the coxal muscle of the tropical cockroach (Nauphoeta cinerea). AB - Adults of warm- and cold-acclimated tropical cockroaches, Nauphoeta cinerea were exposed to low temperatures of 0 or 5 degrees C for various time intervals (hours to days). Development of chilling-injury (defects in crawling and uncoordinated movements) and mortality during the exposure were assessed and correlated with the changes in concentrations of metal ions (Na(+), K(+) and Mg(2+)) in the haemolymph and coxal muscle tissue. Warm-acclimated insects entered chill-coma at both low temperatures. In their haemolymph, the [Na(+)] and [Mg(2+)] linearly decreased and [K(+)] increased with the increasing time of exposure. The rate of concentration changes was higher at 0 than at 5 degrees C. The concentration changes resulted in gradually dissipating equilibrium potentials across the muscle cell membranes. For instance, E(K) decreased from -49.8 to -20.7 mV during 7 days at 5 degrees C. Such a disturbance of ion homeostasis was paralleled by the gradual development of chilling-injury and mortality. Most of the cockroaches showed chilling-injury when the molar ratio of [Na(+)]/[K(+)] in their haemolymph decreased from an initial of 4.4 to 2.1-2.5. In contrast, the cold-acclimated cockroaches did not enter chill-coma. They maintained constant concentrations of ions in their haemolymph, constant equilibrium potentials across muscle cell membranes and the development of chilling-injury was significantly suppressed at 5 degrees C for 7 days. PMID- 16364671 TI - Laminopathies: multisystem dystrophy syndromes. AB - Laminopathies are a heterogeneous group of genetic disorders due to abnormalities in type A lamins and can manifest varied clinical features affecting many organs including the skeletal and cardiac muscle, adipose tissue, nervous system, cutaneous tissue, and bone. Mutations in the gene encoding lamins A and C (LMNA) cause primary laminopathies, including various types of lipodystrophies, muscular dystrophies and progeroid syndromes, mandibuloacral dysplasia, dilated cardiomyopathies, and restrictive dermopathy. The secondary laminopathies are due to mutations in ZMPSTE24 gene which encodes for a zinc metalloproteinase involved in processing of prelamin A into mature lamin A and cause mandibuloacral dysplasia and restrictive dermopathy. Skin fibroblast cells from many patients with laminopathies show a range of abnormal nuclear morphology including bleb formation, honeycombing, and presence of multi-lobulated nuclei. The mechanisms by which mutations in LMNA gene cause multisystem dystrophy are an active area of current investigation. Further studies are needed to understand the underlying mechanisms of marked pleiotropy in laminopathies. PMID- 16364672 TI - Long-term follow-up of Chinese patients who received delayed treatment for 6 pyruvoyl-tetrahydropterin synthase deficiency. AB - OBJECTIVES: 6-Pyruvoyl-tetrahydropterin synthase (PTPS) deficiency is the most important type of BH4 deficiency related to hyperphenylalaninemia. PTPS deficiency may not only cause a typical phenylketonuric phenotype, but is also accompanied by various neurological signs and symptoms due to impaired synthesis of catecholamines and serotonin. Reports of the long-term outcomes of these patients, especially after delayed onset of therapy, are few. STUDY DESIGN: We reviewed the characteristics of 10 PTPS-deficient patients whose treatment onset with tetrahydrobiopterin, L-DOPA, and hydroxytryptophan had been delayed. The relationships among clinical manifestations, biochemical findings, genotypes, and long-term outcomes were analyzed. RESULTS: We classified eight patients as having severe forms, and two as having moderate forms of PTPS deficiency. Improvements in neurological status and intelligence/developmental quotient (IQ/DQ) were observed in all patients, up to approximately 15 years of follow-up. One patient began walking and talking after 4 years of treatment. In patients with severe disease, the mean initial IQ/DQ was 45.40 +/- 13.94, and the final full-scale intelligence quotient (FIQ) score was 62.8 +/- 13.06 (p = 0.042), with a mean increment of 17.4 +/- 5.27 over 15.86 +/- 4.85 years of follow-up. Two patients with moderately severe disease had FIQ increases from 75 to 77 and from 76 to 80 points, respectively. CONCLUSIONS: The administration of neurotransmitters based on clinical response and adverse effects was beneficial in patients whose treatment of PTPS deficiency was delayed. Sustained clinical improvements were observed up to 15 years of follow-up. PMID- 16364673 TI - Diabetes, smoking, and other medical conditions in relation to Parkinson's disease risk. AB - Associations of Parkinson's disease (PD) with diabetes mellitus (DM) and other medical conditions were investigated in a case-control study of 352 newly diagnosed PD case and 484 control subjects. Men with DM had a significantly lower risk of PD, than men without DM (odds ratio (OR) = 0.52, 95% confidence interval (CI) = 0.28, 0.97), whereas the association in women was weaker (OR = 0.80, 95% CI = 0.35, 1.83). PD risk was reduced among male and female smokers, with and without diabetes. However, among diabetics, the PD risk was especially reduced in non-smoking men (OR = 0.09, 95% CI = 0.02, 0.44). Other medical conditions showed no strong relations with PD. PMID- 16364674 TI - Movement disorders and Creutzfeldt-Jakob disease: a review. AB - Movement disorders are reported in a significant number of patients within the course of Creutzfeldt-Jakob disease (CJD). Although myoclonus is more frequent, dystonia, choreoathetosis, tremor, hemiballismus, and atypical parkinsonian syndromes have also been reported. In this review, we report the principal movement disorders associated with CJD and evaluate their correlations with neuroradiological and neuropathological findings that could in fact suggest a basal ganglia dysfunction. Further studies are warranted in order to clarify these correlations. PMID- 16364675 TI - Treatment of levodopa-induced dyskinesias with donepezil. PMID- 16364676 TI - Single-blind clinical trial of psychotherapy for treatment of psychogenic movement disorders. AB - BACKGROUND: PMD are disabling, but lack any generally accepted treatment strategies. DESIGN/METHODS: Ten patients with PMD received treatment with Psychodynamic Psychotherapy. Patients were assessed with psychiatric rating scales and the movement disorder was rated by a blinded rater with the Psychogenic Movement Disorder Rating Scale (PMDRS). RESULTS: Total mean PMDRS (p = 0.0195), total PMDRS function scores (p = 0.0142), Hamilton depression scores (p = 0.009), Beck anxiety scores (p = 0.002), and GAF (p = 0.0083) all improved with psychotherapeutic intervention. CONCLUSIONS: Psychotherapy and appropriate use of adjunct psychiatric medication can be a successful intervention for PMD. PMID- 16364678 TI - Evaluation of proliferative potential in oral lichen planus and oral lichenoid lesions using immunohistochemical expression of p53 and Ki67. AB - A small proportion of cases diagnosed as oral lichen planus (OLP) and oral lichenoid lesions (OLL) can undergo malignant transformation. Some authors, however, stand that only dysplastic lichenoid lesions, not true OLP, have the potential to progress to oral squamous cell carcinoma (OSCC). The histologic diagnosis is a subjective resource and is not always accurate in differentiating OLP from OLL. Thus, this study attempted to evaluate the malignant potential of lesions diagnosed as OLP and as OLL without dysplasia. The Streptavidin-biotin method of immunohistochemistry was used for the staining with p53 and Ki67 in 22 cases of OLP and 27 cases diagnosed as OLL. Ki67 immunoexpression was not statistically different between OLP and OLL (p = 0.353), but, p53 staining showed a significant contrast (p = 0.036). A higher average of staining was detected in the group of OLP. The study showed that apparently a diagnosis of OLP or OLL makes no difference for the patient regarding malignant transformation, although in OLP p53 showed a higher index of expression, probably related to the intensity of inflammatory infiltrate. PMID- 16364677 TI - Stromal syndecan-1 expression is an adverse prognostic factor in oral carcinomas. AB - Syndecan-1, a transmembrane proteoglycan, may exert anti-proliferative effects and promote cell growth by binding various growth factors. Malignant epithelial cells often down-regulate their own syndecan-1 production, and are capable of inducing an aberrant syndecan-1 expression in stromal fibroid cells. Decreased tumor cell syndecan-1 levels in human head and neck squamous cell carcinomas indicate poor prognosis, however, no correlation between stromal syndecan-1 expression and clinical parameters has previously been established. By means of immunohistochemistry, we observed a decrease in tumor cell syndecan-1 reactivity in 33/39 oral carcinoma cases, the degree of which, however, correlated only weakly with the clinical outcome (p = 0.097). Conversely, stromal syndecan-1 positivity proved to be a significant risk factor of recurrence (Cox analysis, p = 0.03) and tumor-specific death (p = 0.023) within a 24-month period after operation. Taken together, stromal expression of syndecan-1 is a reliable factor of adverse prognosis in oral carcinomas. PMID- 16364679 TI - Mathematical models for the control of cystic echinococcosis. AB - Cystic echinococcosis (CE) caused by Echinococcus granulosus is a global public health problem. In many areas the disease is being diagnosed in increasing numbers, whilst in other areas it is re-emerging due to the collapse of public health programmes associated with socio-economic changes. Mathematical models of the transmission dynamics between animals can have an important role to play in developing control options. In particular the parasite is highly endemic in many lower income countries where resources to undertake an intensive control programme that has been successful in wealthy countries, such as New Zealand, are not available. Data from dogs and livestock have been collected and modelled from a number of different countries and regions. In Australia and New Zealand transmission modelling was first developed and these models have been refined using data from the Middle East and central Asia. The model indicates that relatively intense anthelmintic treatment of the dog population will result in a substantial decrease in the parasite population over time and has been supported by the results of control programmes. However, if the newly developed sheep vaccine is included in the control programme, then it should be possible to treat dogs less frequently to achieve the same result. This is due to the potentiating effects of attacking the parasite at two places in its life cycle. This should result in considerable cost savings over the use intensive anthelmintic treatment as the sole method of control. PMID- 16364680 TI - Expression of epithelial cellular adhesion molecule (Ep-CAM) in chronic (necro )inflammatory liver diseases and hepatocellular carcinoma. AB - Epithelial cell adhesion molecule (Ep-CAM) is expressed in a several epithelial tissues and carcinomas, but not on mature hepatocytes. Here, we analysed the expression of Ep-CAM in 230 patients suffering from various liver diseases like chronic hepatitis B and C (HBV and HCV infection), chronic autoimmune hepatitis (AIH), chronic alcoholic liver disease (ALD), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), hereditary hemochromatosis and dysplastic nodules (DNs) as well as hepatocellular carcinomas (HCCs) and cholangiocellular carcinomas (CCCs) by immunohistochemistry. De novo hepatocellular Ep-CAM expression was found in 75.9% of ALD (22/29), 63.6% of HCV (21/33) and 55.6% of each AIH and HBV cases (5/9 and 15/27, respectively). Lower Ep-CAM expression levels were observed for primary sclerosing liver diseases (PBC and PSC) with 25% (3/12) and 7.7% (1/13) of cases. Moreover, only 14.3% of HCCs (9/63) manifested expression, while all CCCs showed strong Ep-CAM expression (5/5). For DNs and hereditary hemochromatosis, Ep-CAM expression was found in 10 and 50% (3/30 and 2/4), respectively. In HBV and HCV, Ep-CAM expression correlated significantly with inflammatory activity as assessed by histological parameters and to the extent of fibrosis. In addition, for HCV also transaminase levels correlated significantly with Ep-CAM expression. Our results indicate that de novo Ep-CAM expression in hepatocytes is frequent in inflammatory liver diseases and is potentially linked to regenerative activity. CCCs and Ep-CAM positive HCCs may represent an attractive target group for Ep-CAM-directed immunotherapies, yet unwanted toxicity may limit the use of such strategies due to Ep-CAM expression in biliary epithelium and several chronic liver diseases such as HBV-and HCV hepatitis. PMID- 16364681 TI - Oxidative stress and hepatitis C viral infection. AB - The involvement of oxidative stress in the pathogenesis of hepatitis and hepatocellular carcinoma has been strongly suggested. Oxidative stress is produced by inflammatory processes that occur in hepatitis via immunological mechanisms. In addition, in hepatitis C virus (HCV) infectious disease, some role has been assigned to viral proteins in the induction of oxidative stress. In the presence of hepatic steatosis, insulin resistance and increased levels of some cytokines, all of which are also induced by viral protein expression, oxidative stress is enhanced in HCV infection. In this sense, the role of oxidative stress in the progression of chronic hepatitis and hepatocarcinogenesis is greater in hepatitis C than in other types of hepatitis such as hepatitis B or autoimmune hepatitis. The additive effects of oxidative stress caused by the inflammatory process and that induced by HCV proteins may, furthermore, exert synergistic effects with alterations in intracellular signaling systems such as mitogen activated protein kinases (MAPK), which are also induced by HCV proteins. These synergistic effects may be responsible for rare characteristics, that is, the high incidence and multicentric nature of hepatocarcinogenesis in HCV infection. PMID- 16364682 TI - Histopathological evaluation of liver biopsy specimens in children with chronic hepatitis B. AB - INTRODUCTION: Histopathological evaluation of the liver remains important diagnostic tool. OBJECTIVE: The aim of this study was to assess inflammatory activity, fibrosis and their correlation to the expression of viral antigens in the liver of children with chronic hepatitis B (CHB) before antiviral treatment. MATERIAL AND METHODS: The study included 190 liver biopsies of children aged 1.5 18 (mean 7.46+/-4.05 years) with CHB. The histopathological examination was based on the modified Knodell system. Additionally, immunomorphological analysis was performed in 125 specimens to detect HBsAg and HBcAg. RESULTS: Necroinflammatory activity was scored for mild in 109 children and moderate in 49. Fibrosis was scored for S1 in 90, S2 in 58 and S3-S4 in seven cases. Positive correlation between grading and staging was observed (chi(2)=77.65, p=0.000002). HBsAg was detected in 62 specimens, while HBcAg was found in the nuclei of 108 samples with cytoplasmic expression in 35-28% cases. No correlation of HBsAg expression to histopathological lesions was established whereas partial correlation of HBcAg expression with inflammatory infiltrate was confirmed. CONCLUSIONS: Progression of liver injury in children with CHB varies in severity. Necroinflammatory activity correlates with fibrosis. Expression of viral antigens is independent of histological changes, however confirms the etiology of liver disease. PMID- 16364683 TI - Divergent activities of interferon-alpha subtypes against intracellular hepatitis C virus replication. AB - BACKGROUNDS: Interferon (IFN)-alpha is represented by several structurally related subtypes that show different antiviral and anti-tumor effects. Here, we analyzed differential effects of IFN-alpha subtypes on intracellular hepatitis C virus (HCV) replication using HCV subgenomic replicon system as a model. METHODS: Huh7 and HeLa cells supporting expression of HCV replicon were treated with various concentrations of five recombinant human IFN-alpha subtypes 1, 2, 5, 8, and 10, and with IFN-alpha con1. The effects of IFNs on various cell-signaling pathways were assayed by using ISRE-, GAS-, AP1-, NF-kappa B-, CRE-, and SRE luciferase reporter plasmids. RESULTS: Each IFN-alpha subtype suppressed HCV replication in a dose-dependent manner. Among them, IFN-alpha8 was the most effective, while IFN-alpha1 was the least effective with 50% inhibitory concentrations of 0.123IU/ml versus 0.375IU/ml, respectively. These differential effects against HCV replication did not correlate with levels of the IFN responsive ISRE or GAS reporter activities, nor they did activate the other reporters, AP1, NF-kappa B, CRE and SRE. CONCLUSION: There were divergent effects of IFN-alpha subtypes against HCV replication that may be through JAK-STAT independent pathways. Exploring further mechanisms of action may elucidate IFN mediated cellular antiviral mechanisms. PMID- 16364684 TI - Altered response to rTMS in patients with Alzheimer's disease. AB - OBJECTIVE: In this study, we tested the excitability of cortical motor areas in patients with Alzheimer's disease. Because repetitive transcranial magnetic stimulation (rTMS) modulates cortical excitability, possibly by inducing a short term increase in synaptic efficacy, we used rTMS to investigate motor cortex excitability in patients with Alzheimer's disease. METHODS: We tested the changes in the size and threshold of motor evoked potential (MEP) and cortical silent period (CSP) duration evoked by focal rTMS delivered in 10 trains of 10 stimuli at 5Hz frequency and 120% rMth intensity in a group of patients with Alzheimer's disease, and age-matched controls. In a further session, rTMS was also delivered at 1Hz frequency (trains of 10 stimuli, 120% rMth). RESULTS: Whereas in control subjects, 5Hz-rTMS elicited normal MEPs that progressively increased in size during the train, in patients, it elicited MEPs that decreased in size. The increase in the duration of the CSP was similar in patients and healthy controls. One hertz rTMS left the MEP amplitude unchanged in patients and healthy controls. CONCLUSIONS: The lack of MEP facilitation reflects an altered response to 5Hz rTMS in patients with Alzheimer's disease. SIGNIFICANCE: Our rTMS findings strongly suggest an altered cortical plasticity in excitatory circuits within motor cortex in patients with Alzheimer's disease. PMID- 16364685 TI - The effect of cannabis use and gender on the visual steady state evoked potential. AB - OBJECTIVE: Cannabis use often induces subjective distortions of perception. However, little work has been done examining the electroencephalographic (EEG) correlates of early sensory processing in cannabis users. The present study therefore examined visual function in cannabis users as assessed via the steady state visual evoked potential. (SSVEP). METHODS: SSVEPs were examined in current cannabis users (n = 17; 59% male; mean age = 23.2 (S.D. = 5.3)) and drug-naive controls (n = 16; 38% male; mean age = 21.3 (S.D. = 3.1)) to periodic photic stimulation presented at 18 and 25 Hz. The visual SSVEP was quantified via spectral power and the phase-locking factor (PLF) at each frequency of stimulation. The transient N160 event-related potential (ERP) was also evaluated at stimulus onset. RESULTS: The results showed that for both frequencies, female subjects in general displayed larger visual SSVEPs. A significant gender X group interaction also occurred at 18 Hz of stimulation, and age of onset of cannabis use positively correlated with 18 Hz spectral power values. Finally, the transient N160 component was shown to be reduced in cannabis users, regardless of gender. CONCLUSIONS: The present study was the first to demonstrate altered visual SSVEPs in cannabis users, and extends previous research demonstrating increased steady state responses in female subjects. While decreased SSVEPs provide initial evidence of altered oscillatory properties in primary visual circuits, reduction of the transient N160 component suggests disruption of later stage visual processing in cannabis users. SIGNIFICANCE: These data provide evidence of cannabinoid modulation of sensory/perceptual function in the visual system, and indicates that cannabis use may affect the oscillatory properties of sensory cortical circuits. PMID- 16364686 TI - Cortical activation with deep brain stimulation of the anterior thalamus for epilepsy. AB - OBJECTIVE: We studied the relation between thalamic stimulation parameters and the morphology, topographic distribution and cortical sources of the cerebral responses in patients with intractable epilepsy undergoing deep brain stimulation (DBS) of the thalamus. METHODS: Bipolar and monopolar stimuli were delivered at a rate of 2 Hz to the anterior (AN, four patients), the dorsomedian (DM, four patients), and the centromedian nucleus (CM, one patient) using the programmable stimulation device (Medtronic ITREL II). Source modeling was carried out by using statistical non-parametric mapping of low-resolution electromagnetic tomography (LORETA) values. RESULTS: All patients demonstrated reproducible time-locked cortical responses (CRs) consisting of a sequence of components with latencies between 20 and 320 ms. The morphology of these CRs, however, was very heterogeneous, depending primarily on the site of stimulation. Following AN stimulation, cortical activation was most prominent in ipsilateral cingulate gyrus, insular cortex and lateral neocortical temporal structures. Stimulation of the DM mainly showed activation of the ipsilateral orbitofrontal and mesial and lateral frontal areas, but also involvement of mesial temporal structures. Stimulation of the CM showed a rather diffuse (though still mainly ipsilateral) increase of cortical activity. The magnitude of cortical activation was positively related to the strength of the stimulus and inversely related to the impedance of the electrode. CONCLUSIONS: The pattern of cortical activation corresponded with the hodology of the involved structures and may underscore the importance of optimal localization of DBS electrodes in patients with epilepsy. SIGNIFICANCE: The method of analyzing sources of CRs could potentially be a useful tool for titration of DBS parameters in patients with electrode contacts in clinically silent areas. Furthermore, the inverse relation of the cortical activation and the impedance of the electrode contacts might suggest that these impedance measurements should be taken into consideration when adjusting DBS parameters in patients with epilepsy. PMID- 16364687 TI - Muscle synergies involved in preparation to a step made under the self-paced and reaction time instructions. AB - OBJECTIVE: To investigate multi-muscle synergies during preparation to making a step and their changes with speed of stepping and method of step initiation, self paced vs. reaction-time. METHODS: Standing healthy persons performed stepping tasks under 3 instructions, 'comfortably, self-paced', 'very quick, self-paced', and 'as fast as possible to a visual signal'. Electromyographic (EMG) signals of 10 postural muscles were recorded and analyzed. The participants also performed standard tasks associated with releasing a load. These trials were used to identify muscle groupings (M-modes) associated with shifts of the center of pressure (COP) and relations between small changes in the M-modes and COP shifts. The framework of the uncontrolled manifold hypothesis was used to partition the EMG variance across stepping trials into two components that kept constant and changed the COP coordinates in the anterior-posterior (AP) and medio-lateral (ML) directions. RESULTS: Muscle synergies stabilizing COP trajectories in both AP and ML directions were observed in both the stepping and supporting legs. The synergies tended to weaken and disappear close to the moment of take-off. They showed only minor differences across the 3 tasks. In contrast, mechanical and EMG characteristics of postural adjustments to stepping were significantly larger and showed higher rates of change during quick steps and steps under the reaction time instruction. CONCLUSIONS: COP shifts associated with preparation to making a step are stabilized by multi-muscle synergies. These synergies are robust across stepping speed and methods of step initiation, self-paced and reaction-time. SIGNIFICANCE: Studies of multi-muscle postural synergies are important for understanding postural disorders in neurological patients and changes in postural strategies with age. PMID- 16364688 TI - Chaperone-related immune dysfunction: an emergent property of distorted chaperone networks. AB - Molecular chaperones (heat shock proteins) are important components of cellular networks, such as protein-protein and gene regulatory networks. Chaperones participate in the folding of immunologically important proteins, presentation of antigens and activation of the immune system. Here, we propose that chaperone related immune dysfunction might be more general than was previously thought. Mutations and polymorphism of chaperones and the regulators of their synthesis, heat shock factor-1, chaperone diseases, sick chaperones and chaperone overload might all affect (mostly impairing) immune responses. PMID- 16364689 TI - Screening and the prevention of gynecologic cancer: endometrial cancer. AB - Over the years, endometrial cancer has remained the most common gynecologic malignancy in the United States. Two categories of endometrial cancer exist: type I and type II. Type I cancers constitute the majority of cases of endometrial cancer, and the risk factors for this type have been studied in greatest detail. These cancers are driven by estrogen, and many of the risk factors are directly or indirectly linked to a state of excessive estrogen. Protective factors seem to be related to conditions that may result in decreased estrogen exposure. Cure rates for endometrial cancer remain high, mainly because of the early stage at which the majority of cases present. Warning signs of abnormal vaginal bleeding or discharge allow detection of these cancers in their early stages. Screening for these cancers is not effective and often leads to additional unnecessary tests; thus, it is not currently recommended in the general population. PMID- 16364690 TI - Screening for ovarian cancer in the general population. AB - Screening for ovarian cancer in the general population presents several unique challenges. Without a clearly identified premalignant state, efforts have focused on detection of early stage disease. Towards this end, investigators have focused on the use of serum markers and transvaginal ultrasound. CA125 determination is the most reliable serum marker in use, and utilization of serial measurements to calculate risk of cancer appears to have greater utility than evaluation of a single value. Multimodality screening focuses on combining serial CA125 measurement with transvaginal ultrasound follow-up for those with abnormal values. Large prospective trials, such as the United Kingdom Collaborative Trial of Ovarian Cancer Screening (UKCTOCS), are currently underway to assess the impact of various screening strategies on mortality, and to evaluate feasibility, acceptability, and morbidity of screening. Future research efforts will undoubtedly focus on promising techniques to examine the serum proteosome for patterns to identify early ovarian cancer. PMID- 16364691 TI - Familial ovarian cancer screening. AB - Familial predisposition accounts for approximately 10% of epithelial ovarian cancer. Identification and management of at-risk families is therefore an important area that bridges gynaecological and clinical genetic practice. The efficacy of screening for ovarian cancer in both high- and low-risk populations is currently of unproven benefit and is associated with some risks related to false-positive results. In contrast to the general population, preventive surgery is a realistic proposition for many women at high-risk. Although prophylactic bilateral salpingo-oophorectomy prevents ovarian and tubal cancer and reduces the risk of breast cancer, this option is unsuitable for women who have yet to complete their families or who are unwilling to undergo surgery. There is therefore a continued demand for screening in the high-risk population. This chapter discusses the limitations of screening, the circumstances under which screening is appropriate and current screening guidelines. Ongoing and future research that should help to provide additional information about this area is also reviewed. PMID- 16364692 TI - Quantitative comparison of intracellular trafficking and nuclear transcription between adenoviral and lipoplex systems. AB - To develop nonviral gene vectors that are sufficient for clinical application, it is necessary to understand why and to what extent nonviral vectors are inferior to viral vectors, which in general show a more efficient transfection activity. This study describes a systematic and quantitative comparison of the cellular uptake and subsequent intracellular distribution (e.g., endosome/lysosome, cytosol, and nucleus) of exogenous DNA transfected by viral and nonviral vectors in living cells, using a combination of TaqMan PCR and a recently developed confocal image-assisted three-dimensionally integrated quantification method. As a model, adenovirus (Ad) and Lipofectamine Plus (LFN) were used for comparison since they are highly potent and widely used viral and nonviral vectors, respectively. The findings indicate that the efficiency of cellular uptake for LFN is significantly higher than that for Ad. Once taken up by a cell, Ad exhibited comparable endosomal escape and slightly higher nuclear transfer efficiency compared with LFN. In contrast, LFN requires 3 orders of magnitude more intranuclear gene copies to exhibit a transgene expression comparable to that of the Ad, suggesting that the difference in transfection efficiency principally arises from differences in nuclear transcription efficiency and not from a difference in intracellular trafficking between Ad and LFN. PMID- 16364693 TI - CNS-directed AAV2-mediated gene therapy ameliorates functional deficits in a murine model of infantile neuronal ceroid lipofuscinosis. AB - The neuronal ceroid lipofuscinoses (Batten disease) are a group of inherited neurodegenerative diseases characterized by the progressive intralysosomal accumulation of autofluorescent material in many cells, visual defects, seizures, cognitive deficits, and premature death. Infantile neuronal ceroid lipofuscinosis (INCL) has the earliest onset ( approximately 1.5 years of age) and is caused by a deficiency in the lysosomal enzyme palmitoyl protein thioesterase-1 (PPT1). Currently there is no effective treatment for children with INCL. In this study, newborn PPT1-deficient mice received two (cortex), four (cortex and hippocampus), or six (cortex, hippocampus, and cerebellum) bilateral intracranial injections of AAV2-PPT1. The AAV-treated animals had localized increases in PPT1 activity, decreased autofluorescent material, improved histologic parameters, and increased brain mass. In addition, the treated animals had dose-dependent improvements in a battery of behavioral tests and improved interictal electroencephalographic tracings. However, there was neither a significant decrease in seizure frequency nor an increase in longevity even in INCL animals receiving six injections. These data suggest that early treatment of INCL using gene transfer techniques can be efficacious. However, higher levels or a broader distribution of PPT1 expression, or both, will be required for more complete correction of this neurodegenerative disease. PMID- 16364694 TI - Development and characterization of polyclonal antibodies against the aryl hydrocarbon receptor protein family (AHR1, AHR2, and AHR repressor) of Atlantic killifish Fundulus heteroclitus. AB - The aryl hydrocarbon receptor (AHR) and AHR repressor (AHRR) proteins regulate gene expression in response to some halogenated aromatic hydrocarbons and polycyclic aromatic hydrocarbons. The Atlantic killifish is a valuable model of the AHR signaling pathway, but antibodies are not available to fully characterize AHR and AHRR proteins. Using bacterially expressed AHRs, we developed specific and sensitive polyclonal antisera against the killifish AHR1, AHR2, and AHRR. In immunoblots, these antibodies recognized full-length killifish AHR and AHRR proteins synthesized in rabbit reticulocyte lysate, proteins expressed in mammalian cells transfected with killifish AHR and AHRR constructs, and AHR proteins in cytosol preparations from killifish tissues. Killifish AHR1 and AHR2 proteins were detected in brain, gill, kidney, heart, liver, and spleen. Antisera specifically precipitated their respective target proteins in immunoprecipitation experiments with in vitro-expressed proteins. Killifish ARNT2 co-precipitated with AHR1 and AHR2. These sensitive, specific, and versatile antibodies will be valuable to researchers investigating AHR signaling and other physiological processes involving AHR and AHRR proteins. PMID- 16364695 TI - Role of O(2)-sensitive K(+) and Ca(2+) channels in the regulation of the pulmonary circulation: potential role of caveolae and implications for high altitude pulmonary edema. AB - High altitude pulmonary edema (HAPE) is a potentially fatal complication in response to exposure to low O(2) at high altitudes. Hypoxia, by causing pulmonary vasoconstriction, increases pulmonary vascular resistance and pulmonary arterial pressure, both of which are features in the pathogenesis of HAPE. Uneven hypoxic pulmonary vasoconstriction is thought to be responsible for increased capillary pressure and leakage, resulting in edema. O(2)-sensitive ion channels are known to play pivotal roles in determining vascular tone in response to hypoxia. K(+), Ca(2+) and Na(+) channels are ubiquitously expressed in both endothelial and smooth muscle cells of the pulmonary microvasculature, subfamilies of which are regulated by local changes in P(O(2)). Hypoxia reduces activity of voltage-gated K(+) channels and down-regulates their expression leading to membrane depolarization, Ca(2+) influx in pulmonary artery smooth muscle cells (by activating voltage-dependent Ca(2+) channels) and vasoconstriction. Hypoxia up regulates transient receptor potential channels (TRPC) leading to enhanced Ca(2+) entry through receptor- and store-operated Ca(2+) channels. Altered enrichment of ion channels in membrane microdomains, in particular in caveolae, may play a role in excitation-contraction coupling and perhaps in O(2)-sensing in the pulmonary circulation and thereby may contribute to the development of HAPE. We review the role of ion channels, in particular those outlined above, in response to low O(2) on vascular tone and pulmonary edema. Advances in the understanding of ion channels involved in the physiological response to hypoxia should lead to a greater understanding of the pathogenesis of HAPE and perhaps in the identification of new therapies. PMID- 16364696 TI - Improved assay for R(-)-apomorphine with application to clinical pharmacokinetic studies in Parkinson's disease. AB - A high performance liquid chromatographic assay for the quantitative determination of apomorphine in human plasma is described. Sample clean-up and concentration was optimised using solid-phase extraction on C18 cartridges, enabling rapid and sensitive determination of apomorphine and potential metabolites. The limit of apomorphine quantification, using fluorescence detection, was 0.5 ng/mL. The assay was stability-indicating, and allowed the detection of analytes in the presence of commonly co-administered anti Parkinsonian drugs. Apomorphine was stable in frozen plasma containing 0.14% (w/v) ascorbic acid for 98 days, and through four freeze-thaw cycles. The assay has been used in clinical pharmacokinetic studies of apomorphine in patients with Parkinson's disease, and in preliminary studies of novel apomorphine delivery devices in volunteers. PMID- 16364697 TI - High sensitivity analysis of oxprenolol in urine by capillary electrophoresis with C18 packed on-line preconcentrator. AB - High sensitivity analysis of oxprenolol in spiked human urine has been performed by capillary zone electrophoresis (CZE) in ammonium formate buffer pH 2.5 using an uncoated capillary with 1cm length C18 on-capillary preconcentrator at the inlet side. The preconcentrator was fabricated in laboratory using the packing method and not encapped C18 5 microm particles as stationary phase material. The packed path was retained into the capillary by sintered stationary phase frits. Before running the CZE analysis, the oxprenolol was eluted from the preconcentrator by injecting a short plug of acetonitrile/water mixtures. With respect to classical CZE, the use of on-line preconcentrator widely increased the method sensitivity allowing the detection of the drug at 0.5 ng/mL (injected concentration). The method showed a linear response in the range of 1-150 ng/mL oxprenolol standard compound. The intra-day repeatability (n = 11) R.S.D. values for migration time, peak area and normalized peak area were 0.72%, 3.96% and 3.66%, respectively, while inter-day repeatability (n = 5 days) R.S.D. values were 2.74%, 9.41% and 9.83%, respectively. The method was successfully applied to the analysis of oxprenolol in extracted urine spiked at 250 pg/mL (oxprenolol LOQ concentration in urine). PMID- 16364698 TI - Quantitative determination of zebularine (NSC 309132), a DNA methyltransferase inhibitor, and three metabolites in murine plasma by high-performance liquid chromatography coupled with on-line radioactivity detection. AB - The metabolism of zebularine (NSC 309132), a novel agent that inhibits DNA methyltransferases, is still uncharacterized. To examine the in vivo metabolism of zebularine, an analytical method was developed and validated (based on FDA guidelines) to quantitate 2-[(14)C]-zebularine and its major metabolites in murine plasma. Zebularine and its metabolites uridine, uracil and dihydrouracil were baseline-separated based on hydrophilic interaction chromatography by using an amino column. The assay was accurate and precise in the concentration ranges of 5.0-100 microg/mL for zebularine, 2.5-50 microg/mL for uridine, 1.0-10 microg/mL for uracil and 0.5-5.0 microg/mL for dihydrouracil. This assay is being used to quantitate zebularine and its metabolites in ongoing pharmacokinetic studies of zebularine. PMID- 16364699 TI - Determination of lapatinib (GW572016) in human plasma by liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS/MS). AB - A sensitive method for the determination of lapatinib (GW572016) in human plasma was developed using high-performance liquid chromatographic separation with tandem mass spectrometric detection. Plasma samples (100 microL) were prepared using solid phase extraction (SPE) columns, and 6.0 microL of the reconstituted eluate was injected onto a Phenomenex CuroSil-PFP 3 mu analytical column (50 mm x 2.0mm) with an isocratic mobile phase. Analytes were detected with a PE SCIEX API 365 LC-MS/MS system at unit (Q1) and low (Q3) resolution in positive multiple reaction monitoring mode (m/z 581 (precursor ion) to m/z 364 (product ion) for lapatinib). The mean recovery for lapatinib was 75% with a lower limit of quantification of 15 ng/mL (S/N=11.3, CV< or =14%). This method was validated over a linear range of 100-10,000 ng/mL, and results from a 5-day validation study demonstrated good within-day and between-day precision and accuracy. This method has been used to measure plasma lapatinib concentrations in a Phase I study in children with cancer. PMID- 16364700 TI - Determination of difenidol hydrochloride by capillary electrophoresis with electrochemiluminescence detection. AB - A novel and sensitive method for the determination of difenidol hydrochloride has been established using capillary electrophoresis coupled with end-column electrogenerated chemiluminescence (ECL) detection, based on the ECL reaction of tris(2,2'-bypyridine)ruthenium(II) (Ru(bpy)(3)(2+)) with the tertiary amino groups of the difenidol analyte. Parameters that affect separation and detection were optimized. Calibration curve was linear over the range from 1 x 10(-6)M to 6 x 10(-5)M with a detection limit of 1 x 10(-7)M (S/N=3). Separation of difenidol hydrochloride from clomifene citrate and lidocaine was achieved using the proposed method. This method was successfully utilized to the assay of the active ingredients of the "difenidol hydrochloride" tablets and to the investigation on the interaction of difenidol hydrochloride with hemoglobin. The number of binding sites and the binding constant were estimated as (11.2 and 2.5) x 10(3)M(-1), respectively. PMID- 16364701 TI - Time discounting and the body mass index. Evidence from the Netherlands. AB - In many Western countries, the relative weight of people -- measured by the body mass index (BMI) -- has increased substantially in recent years, leading to an increasing incidence of overweight and related health problems. As with many forms of risky behavior, it is plausible that overweight is related to the individual discount rate. Increases in credit card debts, the rise in gambling and the development of a more hedonic life style, suggest that the average discount rate has increased over time. An increase in time discounting may be a contributing factor in the rise in BMI. Applying a large set of indicators for the individual discount rate from a Dutch survey, this paper analyzes whether changes in time discounting can account for differences in body mass between individuals at a given point in time and whether changes in the average individual discount rate can explain the remarkable increase in BMI experienced in recent years in the Netherlands. We find some evidence for a link between time discounting and differences in BMI between people, but this relationship depends strongly on the choice of the proxy for the discount rate. Giving our hypothesis the best chance, we analyze the development of the time discounting proxies that are most strongly related to BMI. We find no evidence for a change of these proxies over time. Our main conclusion therefore is that overweight might be related to the way people discount future health benefits, but the increase in BMI is more likely explained by shifts in other parameters that determine the intertemporal decisions regarding the trade-off of current and future health and satisfaction. PMID- 16364702 TI - Doxorubicin coupled to lactosaminated albumin: enhanced drug levels in rat hepatocarcinomas. PMID- 16364703 TI - Young adult outcomes of very-low-birth-weight children. AB - Information on the young adult outcomes of the initial survivors of neonatal intensive care has been reported from the United States, Canada, Australia, Great Britain and other European countries. The studies have varied with regard to whether they were regional or hospital-based, their birth-weight group and gestational age, rates of survival, socio-demographic background, and measures of assessment and types of outcome studied. Despite these differences the overall results reveal that neurodevelopment and growth sequelae persist to young adulthood. Very-low-birth-weight young adults have, with few exceptions, poorer educational achievement than normal-birth-weight controls, and fewer continue with post-high-school study. Rates of employment are, however, similar. There are no major differences in general health status, but the young adults demonstrate poorer physical abilities, higher mean blood pressure and poorer respiratory function. There is no evidence of major psychiatric disorder, although anxiety and depression are reported more often. The young adults report less risk-taking than control populations. They report fairly normal social lives and quality of life. When differences are noted they are usually due to neurosensory disabilities. Longer-term studies are needed to evaluate ultimate educational and occupational achievement. It will also be important to assess the effects of preterm birth, early growth failure and catch-up growth on later metabolic and cardiovascular health. PMID- 16364704 TI - Antenatal care in developing countries: the need for a tailored model. AB - In developed countries, 98% of all women receive prenatal care and 94% give birth under the supervision of skilled healthcare practitioners with timely access to appropriate emergency treatment if complications arise. In contrast, large numbers of pregnant women in Africa and Asia do not receive adequate prenatal care and lack skilled attendance at birth. In developing countries quality of prenatal care is often scarce: models of care adopted in the western world and exported to the developing world have not been monitored early enough to discover their weak points promptly. This blind attitude has transformed antenatal care into an empty and useless ritual, and explains why antenatal care programmes continue to be unsuccessful, being inappropriate to the specific situation. A mix of educational and cultural factors together with persistent lack of resources in a global critical situation all contribute to the poor results of antenatal care programmes. Antenatal care services should be free of charge, planned and implemented within the community, cost-effective, and should yield evidence-based quality care. They should also include information for the patient and family members, provide affordable treatment of existing conditions, and warrant referral for complications. PMID- 16364705 TI - Perinatal audit in low- and high-income countries. AB - Audit is a term used to include case reviews, criterion-based clinical audit, enquiries into maternal mortality and perinatal deaths, and near-miss reviews. The audit cycle consists of identifying cases, collecting information, analysing the results, formulating recommendations, implementing change and re-evaluating practice, and this cycle must be repeated regularly. Implicit in the process are standards against which practice is measured. These standards are becoming increasingly explicit and may be based on hospital protocols or regional or national guidelines. When protocols or guidelines are drawn up, this must be on the basis of multidisciplinary discussion and they need to be regularly updated as new evidence emerges. Audit does not need to be expensive, but it does need the support of all staff, including managers and clinicians. Staff must understand that its purpose is not to identify errors and punish mistakes but to improve clinical care. PMID- 16364706 TI - Midwifery training to improve ante- and perinatal health in low- and middle income countries of the former Soviet Union. AB - Whether in the community or in the hospital, high-quality midwifery care is the preferable model of care for mothers and babies at the first level of care. Countries with professional midwifery care within a supportive system have the best outcomes for mother and babies. The low- and middle-income countries of the former Soviet Union report some of the highest maternal mortality and neonatal mortality in the European region, yet childbirth occurs in institutions with 'skilled attendants' (96-100%). Specific characteristics of maternal and neonatal care in countries of the former Soviet Union include over-medicalization, inappropriate use of technology, unnecessary hospitalizations, and ineffective and/or harmful interventions. This article highlights two midwifery trainings developed specifically to change the maternal and newborn care practices in countries of the former Soviet Union: the Family Centred Maternity Care Training of Trainers and the World Health Organization Essential Antenatal, Perinatal and Postpartum Care Training. PMID- 16364707 TI - The effects of first gestation and lactation on bone metabolism in dairy goats and milk sheep. AB - The goal of the present study was to compare mobilization rate of calcium (Ca) from bone in pregnant and lactating goats and sheep. Blood samples were collected from goats and sheep monthly during pregnancy and at 1, 2, and 4 weeks postpartum (pp) and monthly during lactation until 6 months after parturition. Total bone mineral content (BMC) and total bone mineral density (BMD) were quantified using peripheral quantitative computed tomography at the same intervals as the blood was taken. Bone resorption was assessed by immunoassays quantitating two epitopes of the carboxyterminal telopeptide of type I collagen (ICTP, CTX). Bone formation was estimated by quantifying serum osteocalcin (OC) and bone-specific alkaline phosphatase (bAP). In addition, Ca and 1,25-dihydroxy vitamin D (1,25-VITD) concentrations were determined in serum. Mean ICTP and CTX concentrations of both animal species increased the first week after parturition. By the second week pp, the concentrations of both markers had decreased toward early gestation levels. In contrast, mean OC concentrations continually decreased until the 1st week pp. By the 2nd week pp, the mean concentrations of OC started to increase again. Mean bAP activities decreased during gestation and reached a nadir in the first week pp in goats and 4 weeks pp in sheep. Afterwards, mean bAP activities increased again in goats and sheep. 1,25-VITD concentrations peaked the first week pp and returned to early gestation values thereafter. Total BMC and BMD decreased from the 4th month of pregnancy until the 1st week pp in both species. Afterwards, BMC increased throughout the first month pp in goats and the first 3 months pp in sheep. BMD levels of sheep and goats returned to prepartum levels during lactation. The resorptive phase of bone remodeling is accelerated at parturition and in early lactation and is uncoupled from the process of bone formation. This allows the animal to achieve Ca homeostasis at the expense of bone. Increased bone remodeling during lactation may represent physiological mechanisms to help replace the maternal skeleton lost as the animal adapted to enormously increased Ca losses to the fetus and milk in late gestation and early lactation. PMID- 16364708 TI - Medical care costs of Paget's disease of bone in a privately insured population. AB - INTRODUCTION: Medical care costs are difficult to calculate in diseases such as Paget's disease because they have low detection rates and a wide range of clinical manifestations that commonly occur in aging patient populations. MATERIALS AND METHODS: Using 2001-2002 MarketScan Research databases, this study linked medical claims, prescription records, and encounter data on 2.8 million active and retired employees to create a longitudinal panel with 24 months of observation. Patients with Paget's disease were identified by ICD-9 code 731.0. Matched controls (MC) were identified through an exact match procedure using gender, age, and predicted Medicare costs estimated with a risk adjuster. Diagnostic and expenditure records were extracted for the sample and prevalence rates calculated for 20 conditions with well-documented associations to Paget's disease. Comorbidities and health care costs of Paget's disease patients were compared to those of the MCs, and the differences tested using Chi-square and t tests. RESULTS: Our study identified 244 matched pairs. The average age was 72.7 years; 50.8% were female. Significantly higher comorbidities (P < 0.05) were detected in Paget's disease patients relative to MCs for: pathological fractures (4.9% vs. 0.4%), heart murmurs (3.3% vs. 0.4%), low back pain (19.7% vs. 8.6%), spinal stenosis (16.4% vs. 9.8%), and hearing loss (13.5% vs. 5.7%), respectively. Biannual per patient outpatient costs were significantly higher in Paget's disease patients (Paget's disease $9301 vs. MC $6339, P < 0.05), especially for services associated with physician visits and diagnostic tests. Prescription costs for antiresportive agents and analgesics were also higher (Paget's disease $1115 vs. MC $507, P < 0.05). Inpatient costs (Paget's disease $16,144 vs. MC $21,480) were comparable. CONCLUSION: This study is the first to describe the excessive costs of Paget's disease, based on known patterns of disease expression, evaluation, and treatment. PMID- 16364709 TI - Biglycan deficiency increases osteoclast differentiation and activity due to defective osteoblasts. AB - Bone mass is maintained by a fine balance between bone formation by osteoblasts and bone resorption by osteoclasts. Although osteoblasts and osteoclasts have different developmental origins, it is generally believed that the differentiation, function, and survival of osteoclasts are regulated by osteogenic cells. We have previously shown that the extracellular matrix protein, biglycan (Bgn), plays an important role in the differentiation of osteoblast precursors. In this paper, we showed that Bgn is involved in regulating osteoclast differentiation through its effect on osteoblasts and their precursors using both in vivo and in vitro experiments. The in vivo osteolysis experiment showed that LPS (lipopolisaccharide)-induced osteolysis occurred more rapidly and extensively in bgn deficient mice compared to wild type (WT) mice. To further understand the mechanism of action, we determined the effects of Bgn on 1alpha, 25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3))-induced osteoclast differentiation and bone resorption in an co-culture of calvariae-derived pre-osteoblasts and osteoclast precursors derived from spleen or bone marrow. Time course and dose response experiments showed that tartrate-resistant acid phosphatase-positive multinuclear cells appeared earlier and more extensively in the co-cultures containing calvarial cells from bgn deficient mice than WT mice, regardless of the genotype of osteoclast precursors. The osteoblast abnormality that stimulated osteoclast formation appeared to be independent of the differential production of soluble RANKL and OPG and, instead, due to a decrease in osteoblast maturation accompanied by increase in osteoblastic proliferation. In addition to the imbalance between differentiation and proliferation, there was a differential decrease in secretory leukocyte protease inhibitor (slpi) in bgn deficient osteoblasts treated with 1,25-(OH)(2)D(3). These findings point to a novel molecular factor made by osteoblasts that could potentially be involved in LPS induced osteolysis. PMID- 16364710 TI - Effect of protein supplementation during a 6-month strength and conditioning program on areal and volumetric bone parameters. AB - BACKGROUND: Skeletal loading and proper nutrition are necessary for optimal bone health. The appropriate amount of dietary protein to maximize skeletal health, however, is under constant debate. OBJECTIVE: To determine if 6 months of protein supplementation in conjunction with a strength and conditioning training program improves areal and volumetric bone mineral density (BMD). DESIGN: Fifty-two apparently healthy males and females ages 18-25 years were randomized to protein supplement (PRO, Myoplex, EAS, Inc. Golden CO) containing 280 kcal, 42 g protein, 21 g carbohydrate, and 1.5 g fat) or calorically equivalent carbohydrate control (CS). All subjects participated in a 5 sessions/week strength and conditioning program. Volumetric and areal BMD measurements were made by peripheral quantitative computed tomography (pQCT) of the tibia and whole body DXA. pSSI a measure of torsional bone strength, based on structural and material properties was obtained by pQCT. RESULTS: Measurements at the 20% tibia by pQCT revealed that overall there were significant increases in cortical vBMD (4.3 +/- 1.3 mg/cm(3)), cortical area (1.9 +/- 0.6 cm(2)), cortical thickness (0.05 +/- 0.02 mm) and pSSI (67 +/- 24 mm(3)), and a decrease in endosteal circumference (- 0.5 +/- 0.2 mm) over the intervention period (all, P < 0.05). None of the changes in DXA measures were found to differ by group or sex, there was a trend for a greater increase in whole body BMC among the carbohydrate compared to protein supplemented group and a greater increase among males (16 +/- 8 g) compared to females (-9 +/- 9 g) (P = 0.06). CONCLUSIONS: The results of this study indicate that the consumption of additional protein does not improve measurements of vBMD or bone size during a 6-month strength and conditioning program. Longer duration studies may be necessary to determine the influence of increased dietary protein on bone in young adults. Males and females may have different bone responses to increased protein intake while participating in a strength and conditioning program. PMID- 16364711 TI - Real-time PCR: overview and applications. PMID- 16364712 TI - Error or "act of God"? A study of patients' and operating room team members' perceptions of error definition, reporting, and disclosure. AB - BACKGROUND: Calls abound for a culture change in health care to improve patient safety. However, effective change cannot proceed without a clear understanding of perceptions and beliefs about error. In this study, we describe and compare operative team members' and patients' perceptions of error, reporting of error, and disclosure of error. METHODS: Thirty-nine interviews of team members (9 surgeons, 9 nurses, 10 anesthesiologists) and patients (11) were conducted at 2 teaching hospitals using 4 scenarios as prompts. Transcribed responses to open questions were analyzed by 2 researchers for recurrent themes using the grounded theory method. Yes/no answers were compared across groups using chi-square analyses. RESULTS: Team members and patients agreed on what constitutes an error. Deviation from standards and negative outcome were emphasized as definitive features. Patients and nurse professionals differed significantly in their perception of whether errors should be reported. Nurses were willing to report only events within their disciplinary scope of practice. Although most patients strongly advocated full disclosure of errors (what happened and how), team members preferred to disclose only what happened. When patients did support partial disclosure, their rationales varied from that of team members. CONCLUSIONS: Both operative teams and patients define error in terms of breaking the rules and the concept of "no harm no foul." These concepts pose challenges for treating errors as system failures. A strong culture of individualism pervades nurses' perception of error reporting, suggesting that interventions are needed to foster collective responsibility and a constructive approach to error identification. PMID- 16364713 TI - Adrenocortical carcinoma extending into the inferior vena cava: presentation of a 15-patient series and review of the literature. AB - BACKGROUND: Involvement of the inferior vena cava (IVC) is a controversial risk factor for surgical treatment of adrenocortical carcinoma (ACC). This study aims to assess the outcome of an aggressive surgical policy for ACC extending into the IVC and discuss treatment strategies based on a review of the literature. METHODS: Over a 25-year period, 15 patients were treated for ACC extending into the IVC. The upper limit of the extension was the infrahepatic IVC in 2 patients, retrohepatic IVC in 6, and suprahepatic IVC in 7, including 4 with extension into the right atrium. Seven patients presented with concurrent metastases. The operative technique was thrombectomy (n = 13), partial resection with direct closure (n = 1), and total resection with replacement of the IVC (n = 1). Venous control was achieved by caval clamping alone (n = 4), hepatic vascular exclusion (n = 5), and the use of normothermic cardiopulmonary bypass or hypothermic circulatory arrest (n = 6). RESULTS: Two patients died postoperatively. Ten patients died of metastatic complications at 4 to 31 months. Median survival time was 8 months. Three patients were still alive after 24, 25, and 45 months of follow-up, one of whom was reoperated at 17 months for a local recurrence. No evidence of recurrent intravenous involvement was found during follow-up in any patient in whom complete resection was achieved. CONCLUSIONS: Our findings suggest that surgical treatment can be effective for management of ACC with extension into the IVC. Long-term prognosis is poor owing to delay in diagnosis, frequent associated metastatic disease and lack of effective adjuvant treatment. PMID- 16364714 TI - Contralateral cervical and mediastinal lymph node metastasis in medullary thyroid cancer: systemic disease? AB - BACKGROUND: The current American Joint Committee on Cancer/International Union Against Cancer classification designates cervical and mediastinal lymph nodes as regional lymph nodes. In a unilateral thyroid neoplasm, however, involvement of the contralateral cervical lymph node compartment or the mediastinal lymph node compartment, both of which have been designated "distant" lymph nodes, may serve as a surrogate parameter of distant metastases. METHODS: This institutional series consisted of 105 consecutive patients with medullary thyroid cancer who underwent systematic dissections of both distant lymph node compartments. RESULTS: Thirty-eight patients had no distant lymph node metastasis, 36 patients had involvement of only 1, and 31 patients of both distant lymph node compartments. Significant associations (P < .001) were seen on univariate analysis between the number of involved "distant" lymph node compartments (none, one, or both) and extrathyroidal extension (3%, 33%, and 58%), the number of positive lymph nodes (means of 3, 13, and 33), and distant metastasis (8%, 36%, and 61%). In a multivariate logistic regression model, only involvement of one or both "distant" lymph node compartments (versus no distant lymph node metastasis) remained significantly related to distant metastasis in a dose-dependent fashion. CONCLUSIONS: "Distant" lymph nodes in medullary thyroid cancer should be regarded as nonregional lymph nodes because their involvement is indicative of distant metastasis. PMID- 16364715 TI - Preferable operative age of choledochal dilation types to prevent patients with pancreaticobiliary maljunction from developing biliary tract carcinogenesis. AB - BACKGROUND: Pancreaticobiliary maljunction (PBM), which frequently accompanies choledochal dilation, is a high risk factor for biliary tract (gallbladder, bile duct) carcinoma because of the continuous reflux of pancreatic juice into the biliary tract. The aim of this study was to clarify the preferable operative age in PBM patients for the prevention of biliary tract carcinogenesis, with reference to the dilation types of bile ducts. METHODS: There were 165 PBM patients in total studied, including 92 pediatric patients (< or =15 y) (cystic, 63; spindle-like, 29; nondilation, 0) and 73 adult patients (>15 y) (cystic, 45; spindle-like, 18; nondilation, 10) who underwent operative excision of extrahepatic bile ducts or cholecystectomy. We investigated incidence by age of biliary tract malignancies and the risk according to types of dilation. RESULTS: In the pediatric group, no carcinoma case could be found preoperatively or postoperatively (mean follow-up period, 11.7 y). In the adult group, bile duct carcinomas could be detected in 6 cases of a cystic type (6 of 45; 13.3%) (3 preoperative, 3 postoperative). Among the bile duct carcinoma cases, the youngest patient was a 21-year-old woman who had undergone excision of an extrahepatic bile duct 3 years previously. Gallbladder carcinomas were detected in 16 patients: 3 of 45 cystic (6.7%), 6 of 18 spindle-like (33.3%), and 8 of 10 nondilation (80.0%), in whom the youngest patient was a 41-year-old woman with a spindle-like type. CONCLUSIONS: To prevent biliary tract carcinogenesis in PBM patients, cystic-dilated choledochus should be excised in childhood before the development to a precancerous stage. In spindle-like and nondilation types, cholecystectomy is absolutely necessary in early adulthood before age 40. PMID- 16364716 TI - Immunologic effects of hand-assisted surgery on peritoneal macrophages: comparison to open and standard laparoscopic approaches. AB - BACKGROUND: Exaggerated activation of peritoneal immunity after major abdominal surgery activates peritoneal macrophages (PMs), which may lead to a relative local immunosuppression. Although laparoscopy (L) is known to elicit a smaller attenuation of peritoneal host defenses, compared with open (O) surgery, effects of the hand-assisted (HA) approach have not been investigated to date. METHODS: Eighteen pigs underwent a transabdominal nephrectomy via O, HA, or L approach. PMs were harvested at 4, 12, and 24 hours through an intraperitoneal drain and stimulated in vitro with lipopolysaccharide. The production of interleukin-6 (IL 6) and tumor necrosis factor alpha (TNF-alpha) by the purified macrophage cultures was measured with the use of a standard enzyme-linked immunosorbent assay technique. Statistical comparison was performed by using analysis of variance and Student t test. RESULTS: In vitro lipopolysaccharide-induced IL-6 and TNF-alpha production by PMs increased over the 24-hour period in all 3 groups. Stimulated PMs harvested at 12 and 24 hours postoperatively secreted higher levels of IL-6 in the O group, compared with both the HA group (P = .02, P = .01) and L group (P = .04, P = .001). PMs harvested at 4, 12 and 24 hours postoperatively also produced more TNF-alpha in O group, compared with both the HA group (P = .03, P = .03, and P = .01) and L group (P = .01, P = .05 and P = .03). There was no significant difference between H and L groups in production of either cytokine. CONCLUSIONS: Abdominal surgery attenuates peritoneal host defenses regardless of the surgical approach employed. However, for the first time, we demonstrated that the HA approach, similar to laparoscopy, is superior to open surgery in the degree of PM activation. Overall, in addition to clinical benefits of minimal access, HA surgery may confer an immunologic advantage over laparotomy. PMID- 16364717 TI - Regression of intestinal metaplasia to cardiac or fundic mucosa in patients with Barrett's esophagus submitted to vagotomy, partial gastrectomy and duodenal diversion. A prospective study of 78 patients with more than 5 years of follow up. AB - BACKGROUND: Regression of intestinal metaplasia to cardiac mucosa in patients with Barrett's (BE) esophagus could alter the natural history of BE. OBJECTIVE: To determine the regression of intestinal metaplasia to cardiac mucosa in patients followed more than 5 years after operation, by repeated endoscopy with biopsy. MATERIAL AND METHODS: This prospective study included 78 patients with BE submitted to combined vagotomy, antrectomy (an antireflux procedure), and Roux-en Y gastrointestinal reconstruction with more than 60 months follow up. Patients were divided in 3 groups: (1) 31 with short-segment BE (< or =30 mm length); (2) 42 with long-segment BE (31 to 99 mm length); and (3) 5 with extra-long-segment BE (> or =100 mm). Each patient had at least three endoscopic procedures with multiple biopsies during a mean follow up of 95 months (range, 60-220 months). Acid and duodenal reflux were also evaluated. RESULTS: Sixty-four percent of patients with short segment BE had regression to cardiac mucosa at a mean of 40 months after operation. Sixty-two percent of patients with long segment BE had regression to cardiac mucosa at a mean of 47 months postoperatively. No regression occurred in the 5 patients with extra-long segment BE. In 20% of patients, regression to fundic mucosa occurred between 78 to 94 months after surgery. One patient progressed to low grade dysplasia, but no patient progressed to high-grade dysplasia or adenocarcinoma. Acid and duodenal reflux studies demonstrated that in asymptomatic patients, reflux was abolished; 90% of the patients had a Visick grade of 1 or 2. CONCLUSIONS: Vagotomy and antrectomy combined with duodenal bile diversion abolish acid and duodenal reflux into the distal esophagus in patients with BE, which is accompanied by a regression of BE from intestinal to cardiac or fundic mucosa in about 60% of patients. This regression is time dependent and varies directly with the length of BE. The potential for an antineoplastic effect, especially in young patients with long segment BE, suggests that this operation may become an attractive option as a definitive surgical treatment. Patients with short segment BE submitted to this procedure behave similar to patients submitted to Nissen fundoplication, and therefore in these patients, we do not advocate this complex operation. PMID- 16364718 TI - The effect of morphine on mast cell-mediated mucosal permeability. AB - BACKGROUND: The late phase of post-traumatic multisystem organ failure is associated with sepsis from organisms that normally reside within the gut's lumen. Morphine, a commonly employed analgesic in injured patients, is associated with intestinal stasis, bacterial overgrowth, and translocation when administered to rats. N-formyl-methionyl-leucyl-phenylalanine (FMLP), a toxic product of gram negative organisms, provokes an increase in mucosal permeability when infused into the ileal lumen of this species. The current study was designed to examine the effects of morphine on FMLP perturbation of the mucosal barrier of the ileum of rats and mice to an impermeant macromolecule, dextran 4400. The potential role of mucosal mast cells in the response to either agent alone or in combination was examined. METHODS: Intact and isolated segments of distal ileum of naive and sensitized (Trichinella spiralis and egg albumin) Sprague-Dawley rats were exposed to FMLP with or without morphine or doxantrazole, a mast cell-stabilizing agent. Isolated segments of distal ileum of mast cell-deficient mice also were studied. RESULTS: Mucosal exposure of distal ileal mucosa (intact and isolated, and naive and sensitized) to FMLP was associated with an increase in permeability to dextran 4400, which was completely ablated by morphine and doxantrazole. Sensitization was associated with a prolongation of the FMLP response. Ilea of mast cell-deficient mice (but not their wild type litter mates) were unresponsive to FMLP. CONCLUSIONS: Morphine antagonizes the provocative effect of FMLP on the mucosal barrier to dextran 4400 of the ilea of rats and mice. Intestinal mucosal mast cells play a central role in the process. PMID- 16364719 TI - Attenuation of proinflammatory gene expression and microcirculatory disturbances by endothelin A receptor blockade after orthotopic liver transplantation in pigs. AB - BACKGROUND: Endothelin-1 (ET-1), a very potent mediator of vasoconstriction, leads to microcirculatory disturbances and release of proinflammatory cytokines under pathophysiologic conditions. Our aim was to evaluate the effect of a selective ET(A)-receptor antagonist (ET(A)-RA) on cold ischemia/reperfusion (I/R) injury in a pig model. METHODS: Twenty pigs revealed orthotopic liver transplantation. The animals were randomized into 2 groups: control pigs received isotonic saline; the treated group received the selective ET(A)-RA BSF 208075 at the beginning of reperfusion. On postoperative days 4 and 7, animals were re laparotomized to obtain tissue specimens. Liver tissue samples were collected and quantitative mRNA expression for prepro-ET-1, ET(A) receptor, pro-IL-1beta, pro IL-6, pro-TNF-alpha, and endothelial nitric oxide synthase was analyzed using the TaqMan system. Additionally, immunohistochemical analysis for ET-1 was performed. Hepatic microcirculation was evaluated by laser Doppler flow measurement and partial pressure of oxygen and carbon dioxide measurements with the Paratrend sensor. Postischemic liver damage was monitored by measurement of liver enzymes and by histologic analysis using a semiquantitative scoring classification. RESULTS: Treatment with the ET(A)-RA significantly reduced the severity of I/R injury evidenced by lower serum AST, ALT and GLDH. Analysis of partial pressure of oxygen and blood flow revealed a significant improvement of capillary perfusion and blood flow in the treated group and was associated with a relevant reduction of tissue injury. One hour after reperfusion, quantitative RT-PCR revealed significantly lower expression of prepro-ET-1, ET(A) receptor, endothelial nitric oxide synthase, pro-TNF-alpha, pro-IL-1beta and pro-IL-6 in the therapy group. Immunohistochemical analysis demonstrated significantly reduced ET-1 immunostaining after therapy. Histologic investigation suggested less tissue damage in treated animals. CONCLUSIONS: Treatment with the selective ET(A)-RA BSF 208075 has protective effects on microcirculation after liver transplantation. ET(A)-RA not only affects the expression of vasoactive genes, but also decreases gene expression of proinflammatory cytokines such as TNF alpha, IL-1beta and IL-6. PMID- 16364720 TI - Altered growth patterns of colorectal liver metastases after thermal ablation. AB - BACKGROUND: Thermal ablation by radiofrequency or laser is used increasingly for the treatment of colorectal liver metastases. Recurrence after thermal ablation is common and occurs both locally and at distant sites. One possible cause of this recurrence may be a result of growth stimulation of micrometastases in the remaining liver. This study examined the impact of thermal ablation on growth patterns of hepatic micrometastases. METHODS: Colorectal liver metastases were induced in male CBA-strain mice via an intrasplenic injection of a murine-derived cancer cell line. Subtotal thermal ablation of the left posterior lobe of the liver (30% of total liver volume) was performed by neodymium yttrium-aluminum garnet laser 7 days after induction of metastases. The distribution, number, cross-sectional diameter, volume, and proliferation rate of established neoplasms were compared with controls at 21 days after tumor induction. The effect of thermal ablation of 7% of the total liver volume by laser on the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor 2 (FGF 2), transforming growth factor beta, and cellular proliferation (Ki-67 antigen) adjacent to the ablated site was assessed by immunohistochemistry in separate groups of animals at specific time points after therapy. RESULTS: Thermal ablation did not alter the overall volume, number, size, and proliferation rate of neoplasms 21 days after laser ablation. There were no extrahepatic metastases after therapy. The number of neoplasms in the regenerated posterior lobe was equivalent to control despite subtotal ablation (29 +/- 2 vs 27 +/- 2; P = NS). A greater amount of metastases occupied the regenerated thermal-ablated lobe compared with controls (55% +/- 4% vs 29% +/- 3%; P < .04). Thermal ablation stimulated liver proliferation adjacent to the treatment site at 12 hours compared with untreated controls. Stimulation peaked at 72 hours (20% +/- 1% vs 1% +/- 1%; P < .001) and persisted to 21 days after therapy. FGF-2 and VEGF expression increased in liver tissue adjacent to the ablation site compared with baseline, peaking at 12 hours (112% +/- 2% vs 102% +/- 1%; P < .001) and 72 hours (114% +/- 2% vs 101% +/- 1%; P < .001), respectively. CONCLUSIONS: Thermal ablation promotes the progression of micrometastases to form macroscopically detectable neoplasms in treated regenerating liver. This effect may relate to an increased expression of VEGF and FGF-2 adjacent to the treatment site. PMID- 16364721 TI - Insulin prevents oxidant-induced endothelial cell barrier dysfunction via nitric oxide-dependent pathway. AB - BACKGROUND: The rigorous maintenance of normoglycemia by the administration of insulin is beneficial to critically ill patients. Because insulin induces endothelial nitric oxide (NO) release, and the constitutive release of NO maintains normal microvascular permeability, the authors postulated that insulin would prevent peroxide (H(2)O(2))-induced endothelial barrier dysfunction, an effect dependent on endothelial NO synthase (eNOS) activity. METHODS: Murine lung microvascular endothelial cells (LMEC) grown to confluence on 8 micro pore polyethylene filters were exposed to media (control), H(2)O(2) (20 to 500 micromol/L), insulin (1 to 1,000 nmol/L) or insulin (100 nmol/L) + H(2)O(2) (10( 4)mol/L). Endothelial monolayer permeability was quantitated by measuring the transendothelial electrical resistance at 15-minute intervals for 120 minutes. Other cells were exposed to H(2)O(2) and insulin after pretreatment with a NO scavenger (PTIO), an eNOS inhibitor (L-NIO), or a phosphoinositol-3-kinase inhibitor (LY-294002). RESULTS: H(2)O(2) caused a concentration- and time dependent reduction in electrical resistance consistent with an increase in monolayer permeability. This effect was prevented by insulin. Inhibiting NO release (L-NIO, LY-294002) or scavenging NO (PTIO) abolished this protective effect. CONCLUSIONS: These data suggest that insulin may modulate endothelial barrier function during oxidant stress by inducing the release of NO. PMID- 16364722 TI - Microarray analysis of the differential effects of open and laparoscopic surgery on murine splenic T-cells. AB - BACKGROUND: Surgical trauma depresses cell-mediated immunity of a duration and magnitude proportional to the degree of injury. However, the cellular mechanism underlying this effect is poorly understood. Microarrays were used to survey gene expression in murine splenic T-cells after pneumoperitoneum and laparotomy. METHODS: C3H/HeJ mice were assigned randomly to undergo anesthesia alone, sham laparotomy, or CO(2) pneumoperitoneum and sacrificed 12 or 24 hours later. RNA was isolated from purified splenic T-cells and hybridized to Affymetrix oligonucleotide microarrays. RESULTS: Relative to anesthesia, 116 genes after pneumoperitoneum and 398 genes after laparotomy showed a > or =2-fold change in expression at 12 hours. One hundred thirty-two genes after pneumoperitoneum and 157 genes after laparotomy met those criteria at 24 hours. Comparing surgical modalities, 177 genes were increased and 15 decreased > or =2-fold after laparotomy relative to pneumoperitoneum at 12 hours, compared with 44 and 5 genes respectively at 24 hours. Expression changes for 8 genes were validated by quantitative real-time polymerase chain reaction. CONCLUSIONS: Laparotomy and pneumoperitoneum alter splenic T-cell gene expression, with the most extensive changes occurring 12 hours after laparotomy. This study is one of the first comprehensive genomic studies of the molecular effects of surgical manipulation on immune function. The genes identified are potential targets for modulating the immune response to surgery. PMID- 16364723 TI - Expression of cytokeratin-20 in pancreatic cancer: an indicator of poor outcome after R0 resection. AB - BACKGROUND: After complete removal of the neoplasm (R0 resection), approximately 80% of pancreatic cancer patients will die of the disease within 5 years. The expression panel of cytokeratins (CK) is linked closely with cell differentiation. The aim of the study was to investigate the expression of CK-20 in pancreatic cancer tissue and to correlate CK-20 expression with survival in R0 resected pancreatic cancer patients. METHODS: Tissue samples of 63 patients with pancreatic cancer were subjected to CK-20 reverse-transcription polymerase chain reaction. Thirty-four of 63 patients underwent R0 resection and were followed-up for survival statistics. From these 34 patients, 26 (76%) neoplasms were CK-20 positive and 8 (24%) samples were CK-20 negative. The mean follow-up period for the entire group was 17 months (range, 4-36 mo), the follow-up period in censored patients was 23 months (range, 10-36 mo). RESULTS: In the R0-resected group, 3 of 8 (38%) patients with CK-20-negative neoplasms, and 16 of 26 (62%) patients with CK-20-positive neoplasms (P = .15) died of recurrent disease. The median survival time of patients with CK-20-positive neoplasms was 13 months (range, 4-36), the median survival in R0-resected patients with CK-20-negative neoplasm was 26 months (range, 13-35; P = .06). The survival difference observed in patients with CK-20-negative neoplasms could not be attributed to intergroup variations in tumor stage or tumor grade. CONCLUSIONS: A majority of primary ductal pancreatic adenocarcinomas express CK-20. This seems to be associated with poorer survival in R0-resected patients. Our data suggest that ductal pancreatic adenocarcinomas negative for CK-20 constitute a subgroup of patients showing a more favorable disease outcome. The expression of CK-20 in resected pancreatic cancer may be of interest as a prognostic parameter. PMID- 16364724 TI - The transmanubrial approach: a new operative approach to cervicothoracic neuroblastoma in children. AB - BACKGROUND: Cervicothoracic neuroblastoma originates from the cervical sympathetic nerves and ganglia and thus presents a problem when dissecting the vascular and nervous elements of the subclavian region. The standard operation is based on thoracotomy or dual cervicotomy/thoracotomy, but these approaches do not provide optimal control of the subclavian vessels. We report our experience in children with cervicothoracic neuroblastoma by using a technique usually performed for apical lung cancer. METHODS: Four patients with localized cervicothoracic neuroblastoma with no N-myc amplification were resected after chemotherapy by this approach. The anatomic evaluation was performed preoperatively with angio-magnetic resonance imaging. This transmanubrial approach, performed through a manubrial L-shaped transection and first costal cartilage resection, affords excellent access to the subclavian region with safe control of the vessels and nerves and exposure of the first 4 thoracic intervertebral foramina. RESULTS: Removal of more than 90% of the tumor was possible in all cases. The postoperative course was uneventful in 3 cases, and the fourth patient with a left-sided tumor had a transient chylothorax. No recurrence occurred with a follow-up period of 8 to 32 months. CONCLUSIONS: The transmanubrial approach is an osteomuscular-sparing technique that seems particularly suitable for the treatment of these tumors, which require a resection that is as complete as possible to avoid postoperative chemotherapy and tumor relapse. PMID- 16364725 TI - Sergei S. Yudin: an untold story. PMID- 16364726 TI - Radiofrequency ablation of a liver metastasis complicated by extensive liver necrosis and sepsis caused by gas gangrene. PMID- 16364727 TI - Posterior perineal hernia--an unusual surgical complication! PMID- 16364728 TI - A large stone detected in Roux-en-Y jejunal limb 20 years after excision of congenital choledochal cyst. PMID- 16364730 TI - One hundred years of poliovirus pathogenesis. AB - Poliovirus was first isolated nearly 100 years ago in a landmark experiment that established the viral etiology of poliomyelitis. This discovery stimulated investigation of the pathogenesis of poliomyelitis in many laboratories. Nearly 50 years later, when two effective poliovirus vaccines were developed, the impetus to study poliovirus pathogenesis waned. The identification of the cell receptor for poliovirus, CD155, and its use in the development of transgenic mice susceptible to poliovirus revived interest in understanding how the virus causes disease. Experiments in CD155 transgenic mice have provided new information on the initial sites of virus replication in the host, how the virus spreads to the central nervous system through the blood and by axonal transport, the determinants of viral tropism, and the basis for the attenuation phenotype of the Sabin vaccine strains. Despite these advances, our understanding of poliovirus pathogenesis is still incomplete. The dilemma is not how to answer the remaining questions, but whether there will be sufficient time to do so before global eradication of poliomyelitis leads to cessation of research on the disease. PMID- 16364731 TI - Different receptors binding to distinct interfaces on herpes simplex virus gD can trigger events leading to cell fusion and viral entry. AB - One of the herpes simplex virus envelope glycoproteins, designated gD, is the principal determinant of cell recognition for viral entry. Other viral glycoproteins, gB, gH and gL, cooperate with gD to mediate the membrane fusion that is required for viral entry and cell fusion. Membrane fusion is triggered by the binding of gD to one of its receptors. These receptors belong to three different classes of cell surface molecules. This review summarizes recent findings on the structure and function of gD. The results presented indicate that gD may assume more than one conformation, one in the absence of receptor, another when gD is bound to the herpesvirus entry mediator, a member of the TNF receptor family, and a third when gD is bound to nectin-1, a cell adhesion molecule in the immunoglobulin superfamily. Finally, information and ideas are presented about a membrane-proximal region of gD that is required for membrane fusion, but not for receptor binding, and that may have a role in activating the fusogenic activity of gB, gH and gL. PMID- 16364732 TI - Avian sarcoma and leukosis virus-receptor interactions: from classical genetics to novel insights into virus-cell membrane fusion. AB - For over 40 years, avian sarcoma and leukosis virus (ASLV)-receptor interactions have been employed as a useful model system to study the mechanism of retroviral entry into cells. Pioneering studies on this system focused upon the genetic basis of the differential susceptibilities of different lines of chickens to infection by distinct subgroups of ASLV. These studies led to the definition of three distinct autosomal recessive genes that were predicted to encode cellular receptors for different viral subgroups. They also led to the concept of viral interference, i.e. the mechanism by which infection by one virus can render cells resistant to reinfection by other viruses that use the same cellular receptor. Here, we review the contributions that analyses of the ASLV-receptor system have made in unraveling the mechanisms of retroviral entry into cells and focus on key findings such as identification and characterization of the ASLV receptor genes and the subsequent elucidation of an unprecedented mechanism of virus-cell fusion. Since many of the initial findings on this system were published in the early volumes of Virology, this subject is especially well suited to this special anniversary issue of the journal. PMID- 16364733 TI - Paramyxovirus membrane fusion: lessons from the F and HN atomic structures. AB - Paramyxoviruses enter cells by fusion of their lipid envelope with the target cell plasma membrane. Fusion of the viral membrane with the plasma membrane allows entry of the viral genome into the cytoplasm. For paramyxoviruses, membrane fusion occurs at neutral pH, but the trigger mechanism that controls the viral entry machinery such that it occurs at the right time and in the right place remains to be elucidated. Two viral glycoproteins are key to the infection process-an attachment protein that varies among different paramyxoviruses and the fusion (F) protein, which is found in all paramyxoviruses. For many of the paramyxoviruses (parainfluenza viruses 1-5, mumps virus, Newcastle disease virus and others), the attachment protein is the hemagglutinin/neuraminidase (HN) protein. In the last 5 years, atomic structures of paramyxovirus F and HN proteins have been reported. The knowledge gained from these structures towards understanding the mechanism of viral membrane fusion is described. PMID- 16364734 TI - Class II virus membrane fusion proteins. AB - Enveloped animal viruses fuse their membrane with a host cell membrane, thus delivering the virus genetic material into the cytoplasm and initiating infection. This critical membrane fusion reaction is mediated by a virus transmembrane protein known as the fusion protein, which inserts its hydrophobic fusion peptide into the cell membrane and refolds to drive the fusion reaction. This review describes recent advances in our understanding of the structure and function of the class II fusion proteins of the alphaviruses and flaviviruses. Inhibition of the fusion protein refolding reaction confirms its importance in fusion and suggests new antiviral strategies for these medically important viruses. PMID- 16364735 TI - Poxvirus entry and membrane fusion. AB - The study of poxvirus entry and membrane fusion has been invigorated by new biochemical and microscopic findings that lead to the following conclusions: (1) the surface of the mature virion (MV), whether isolated from an infected cell or by disruption of the membrane wrapper of an extracellular virion, is comprised of a single lipid membrane embedded with non-glycosylated viral proteins; (2) the MV membrane fuses with the cell membrane, allowing the core to enter the cytoplasm and initiate gene expression; (3) fusion is mediated by a newly recognized group of viral protein components of the MV membrane, which are conserved in all members of the poxvirus family; (4) the latter MV entry/fusion proteins are required for cell to cell spread necessitating the disruption of the membrane wrapper of extracellular virions prior to fusion; and furthermore (5) the same group of MV entry/fusion proteins are required for virus-induced cell-cell fusion. Future research priorities include delineation of the roles of individual entry/fusion proteins and identification of cell receptors. PMID- 16364736 TI - Late budding domains and host proteins in enveloped virus release. PMID- 16364737 TI - Filovirus assembly and budding. AB - Filoviruses belong to the order of negative-stranded non-segmented RNA viruses and are classified into two genera, Ebola and Marburg viruses. They have a characteristic filamentous shape, which is largely determined by the matrix protein VP40. Although VP40 is the main driving force for assembly and budding from the host cell, the production of infectious virus involves an intricate interplay between all viral structural proteins in addition to cellular factors, e.g., those that normally function in multi-vesicular body biogenesis. As a consequence, assembly and budding steps are defined to specific cellular compartments, and the recent progress in understanding how the different components are assembled into stable enveloped virus particles is reviewed. PMID- 16364738 TI - Hepatitis delta virus. AB - Hepatitis delta virus (HDV) is a sub-viral agent that is dependent for its life cycle on hepatitis B virus (HBV). The help it obtains from HBV is limited to the sharing of envelope proteins. These proteins are needed to facilitate the assembly of the HDV genome into new virus particles, and in turn, to allow the attachment and entry of HDV into new host cells. In other respects, the replication of the small single-stranded circular RNA genome of HDV is independent of HBV. HDV genome replication produces two forms of a RNA-binding protein known as the long and small delta antigens (Ag). All other proteins needed for HDV genome replication, especially the RNA-directed RNA polymerase activity, are provided by the host cell. This mini-review article is a mixture of personal perspective and speculations about the future of HDV research. It starts with a brief overview of HDV and its replication, notes some of the major unresolved questions, and directs the interested reader to more detailed reviews. PMID- 16364739 TI - Conserved molecular systems of the Baculoviridae. AB - Although the Baculoviridae are a large and diverse family of viruses, they are united by a number of shared features that form the basis for their unique life cycle. These include the mechanism of cell entry, genome replication and processing, and late and very late gene transcription. In this review, the molecular systems that are conserved within the Baculoviridae and that are responsible these processes are described. PMID- 16364740 TI - Retroviral infection of non-dividing cells: old and new perspectives. AB - The dependence of retroviral replication on cell proliferation was described as early as 1958, although different classes of retroviruses are able to infect non dividing cells with different efficiencies. For example, the human immunodeficiency virus (HIV) and other lentiviruses infect most non-dividing cells nearly as well as dividing cells, while the gammaretroviruses such as the murine leukemia virus (MLV) cannot infect non-dividing cells, and other retroviruses have intermediate phenotypes. One exception to the ability of HIV to infect non-dividing cells involves resting CD4+ T cells in vitro where there are multiple restrictions. However, recent data show that there is massive infection of non-activated CD4+ T cell during acute infection which suggests that the situation is different in vivo. Finally, much work trying to explain the difference between HIV and MLV in non-dividing cells has focused on describing the ability of HIV to enter the nucleus during interphase. However, we suggest that events in the viral life-cycle other than nuclear import may be more important in determining the ability of a given retrovirus to infect non-dividing cells. PMID- 16364741 TI - Structural disorder within the replicative complex of measles virus: functional implications. AB - Measles virus belongs to the Paramyxoviridae family within the Mononegavirales order. Its non-segmented, single stranded, negative sense RNA genome is encapsidated by the nucleoprotein (N) to form a helical nucleocapsid. This ribonucleoproteic complex is the substrate for both transcription and replication. The RNA-dependent RNA polymerase binds to the nucleocapsid template via its co-factor, the phosphoprotein (P). In this review, we summarize the main experimental data pointing out the abundance of structural disorder within measles virus N and P. We also describe studies indicating that structural disorder is a widespread property in the replicative complex of Paramyxoviridae and, more generally, of Mononegavirales. The functional implications of structural disorder are also discussed. Finally, we propose a model where the flexibility of the disordered N and P domains allows the formation of a tripartite complex (N degrees-P-L) during replication, followed by the delivery of N monomers to the newly synthesized genomic RNA chain. PMID- 16364742 TI - Viral persistence: parameters, mechanisms and future predictions. AB - For a virus to persist, it must actively curtail the host's antiviral immune response. Here, we review the conceptual basis by which this can occur and discuss the subsequent fate of differentiated cells infected over long periods of time. We also consider how the compromised antiviral immune response can be revigorated or replaced with a potent response that purges the virus and thereby terminates persistent infection. PMID- 16364743 TI - The interferon response circuit: induction and suppression by pathogenic viruses. AB - Type I interferons (IFN-alpha/beta) are potent antiviral cytokines and modulators of the adaptive immune system. They are induced by viral infection or by double stranded RNA (dsRNA), a by-product of viral replication, and lead to the production of a broad range of antiviral proteins and immunoactive cytokines. Viruses, in turn, have evolved multiple strategies to counter the IFN system which would otherwise stop virus growth early in infection. Here we discuss the current view on the balancing act between virus-induced IFN responses and the viral counterplayers. PMID- 16364744 TI - Phosphoinositide 3-kinase: from viral oncoprotein to drug target. AB - The catalytic subunit p110alpha of the phosphoinositide 3-kinase (PI3K) and the serine-threonine protein kinase Akt have been extensively studied as retroviral oncoproteins. The experimental tools developed with the retroviral vectors are now being applied to PI3K mutations in human cancer. The most frequently occurring mutants of p110alpha are oncogenic in vitro and in vivo, show gain of enzymatic function, activate Akt, and their oncogenic activity is sensitive to rapamycin. The related isoforms p110beta, gamma and delta induce oncogenic transformation as wild-type proteins. Mutated p110alpha proteins are ideal drug targets. Identification of small molecule inhibitors that specifically target these mutant proteins is a realistic and urgent goal. PMID- 16364745 TI - Near death experiences: poxvirus regulation of apoptotic death. AB - Apoptosis, or programmed cell death, plays a critical role in the elimination of virus-infected cells. As a result, a growing number of viruses encode numerous potent anti-apoptotic proteins to counteract apoptosis in an effort to prolong their own survival. This review describes the numerous mechanisms by which poxviruses inhibit apoptosis thereby modulating life and death of the cell. PMID- 16364746 TI - Interaction of viruses with the mammalian RNA interference pathway. AB - It has been known for some time that plants and insects use RNA interference (RNAi) as nucleic acid-based immunity against viral infections. However, it was unknown whether mammalian cells employ the RNA interference pathway as an antiviral mechanism as well. Over the past years, it has become clear that a variety of viruses, first in plants but recently in insect and mammalian viruses as well, encode suppressors of the RNAi pathway arguing for an antiviral role of this machinery. More recent findings have revealed that certain viruses encode their own microRNAs or microRNA-like RNA molecules, which are processed by the mammalian RNAi machinery. Furthermore, host-encoded microRNAs have been shown to both silence and enhance intracellular levels of viral RNAs. These findings argue that interactions between the RNAi pathway and viral genomes can profoundly affect the outcomes of the viral life cycles and contribute to the pathogenic signatures of the infectious agents. PMID- 16364747 TI - Silencing suppression by geminivirus proteins. AB - RNA silencing is an RNA-directed gene regulatory system that is present in a wide range of eukaryotes, and which functions as an antiviral defense in plants. Silencing pathways are complex and partially overlapping, but at least three basic classes can be distinguished: cytoplasmic RNA silencing (or post transcriptional gene silencing; PTGS) mediated by small interfering RNAs (siRNAs), silencing mediated by microRNAs (miRNAs), and transcriptional gene silencing (TGS) mediated by siRNA-directed methylation of DNA and histone proteins. Recent advances in our understanding of different geminivirus silencing suppressors indicate that they can affect all three pathways, suggesting that multiple aspects of silencing impact geminivirus replication. PMID- 16364748 TI - Plant viral movement proteins: agents for cell-to-cell trafficking of viral genomes. AB - Plants viruses spread throughout their hosts using a number of pathways, the most common being movement cell to cell through plasmodesmata (PD), unique intercellular organelles of the plant kingdom, and between organs by means of the vascular system. Pioneering studies on plant viruses revealed that PD allow the cell-to-cell trafficking of virally encoded proteins, termed the movement proteins (MPs). This non-cell-autonomous protein (NCAP) pathway is similarly employed by the host to traffic macromolecules. Viral MPs bind RNA/DNA in a sequence nonspecific manner to form nucleoprotein complexes (NPC). Host proteins are then involved in the delivery of MPs and NPC to the PD orifice, and a role for the cytoskeleton has been implicated. Trafficking of NCAPs through the PD structure involves three steps in which the MP: (a) interacts with a putative PD docking complex, (b) induces dilation in the PD microchannels, and (c) binds to an internal translocation system for delivery into the neighboring cytoplasm. Viral genera that use this NCAP pathway have evolved a combination of a MP and ancillary proteins that work in concert to enable the formation of a stable NPC that can compete with endogenous NCAPs for the PD trafficking machinery. Incompatible MP-host protein interactions may underlie observed tissue tropisms and restricted infection domains. These pivotal discoveries are discussed in terms of the need to develop a more comprehensive understanding of the (a) three dimensional structure of MPs, (b) PD supramolecular complex, and (c) host proteins involved in this cell-to-cell trafficking process. PMID- 16364750 TI - Subviral agents associated with plant single-stranded DNA viruses. AB - Begomoviruses (family Geminiviridae) are responsible for many economically important crop diseases worldwide. The majority of these diseases are caused by bipartite begomovirus infections, although a rapidly growing number of diseases of the Old World are associated with monopartite begomoviruses. With the exception of several diseases of tomato, most of these are caused by a monopartite begomovirus in association with a recently discovered essential satellite component (DNA-beta). These begomovirus/satellite disease complexes are widespread and diverse and collectively infect a wide variety of crops, weeds and ornamental plants. Non-essential subviral components (DNA-1) originating from nanoviruses are frequently associated with these disease complexes, and there are tantalizing hints that further novel satellites may also be associated with some begomovirus diseases. DNA-beta components can be maintained in permissive plants by more than one distinct begomovirus, reflecting less stringent requirements for trans-replication that will undoubtedly encourage diversification and adaptation as a consequence of component exchange and recombination. In view of their impact on agriculture, there is a pressing need to develop a more comprehensive picture of the diversity and distribution of the disease complexes. A greater understanding of how they elicit the host response may provide useful information for their control as well as an insight into plant developmental processes. PMID- 16364751 TI - Yeast as a model host to dissect functions of viral and host factors in tombusvirus replication. AB - RNA replication is the central process during the infectious cycles of plus stranded RNA viruses. Development of yeast as a model host and powerful in vitro assays with purified replicase complexes, together with reverse genetic approaches make tombusviruses, small plant RNA viruses, excellent systems to study fundamental aspects of viral RNA replication. Accordingly, in vitro approaches have led to the identification of protein-RNA interactions that are essential for template selection for replication and assembly of the functional viral replicase complexes. Moreover, genome-wide approaches and proteomics analyses have identified a new set of host proteins that affected tombusvirus replication. Overall, rapid progress in tombusvirus replication has revealed intriguing and complex nature of virus-host interactions, which make robust replication of tombusviruses possible. The knowledge obtained will likely stimulate development of new antiviral methods as well as other approaches that could make tombusviruses useful tools in biotechnological applications. PMID- 16364749 TI - Translational control in positive strand RNA plant viruses. AB - The great variety of genome organizations means that most plant positive strand viral RNAs differ from the standard 5'-cap/3'-poly(A) structure of eukaryotic mRNAs. The cap and poly(A) tail recruit initiation factors that support the formation of a closed loop mRNA conformation, the state in which translation initiation is most efficient. We review the diverse array of cis-acting sequences present in viral mRNAs that compensate for the absence of a cap, poly(A) tail, or both. We also discuss the cis-acting sequences that control translation strategies that both amplify the coding potential of a genome and regulate the accumulations of viral gene products. Such strategies include leaky scanning initiation of translation of overlapping open reading frames, stop codon readthrough, and ribosomal frameshifting. Finally, future directions for research on the translation of plant positive strand viruses are discussed. PMID- 16364752 TI - Fifty-three years since Hershey and Chase; much ado about pressure but which pressure is it? AB - The events that occur at the initiation of phage infection are discussed, from adsorption through DNA ejection from the virion into the cell. A new model for DNA translocation is described that not only overcomes difficulties associated with previous models of DNA ejection but also provides a mechanism by which both single-stranded genomes and internal phage proteins can be transported from the virion into the cell cytoplasm. PMID- 16364753 TI - Replication-defective viruses as vaccines and vaccine vectors. AB - The classical viral vaccine approaches using inactivated virus or live-attenuated virus have not been successful for some viruses, such as human immunodeficiency virus or herpes simplex virus. Therefore, new types of vaccines are needed to combat these infections. Replication-defective mutant viruses are defective for one or more functions that are essential for viral genome replication or synthesis and assembly of viral particles. These viruses are propagated in complementing cell lines expressing the missing gene product; however, in normal cells, they express viral gene products but do not replicate to form progeny virions. As vaccines, these mutant viruses have advantages of both classical types of viral vaccines in being as safe as inactivated virus but expressing viral antigens inside infected cells so that MHC class I and class II presentation can occur efficiently. Replication-defective viruses have served both as vaccines for the virus itself and as a vector for the expression of heterologous antigens. The potential advantages and disadvantages of these vaccines are discussed as well as contrasting them with single-cycle mutant virus vaccines and replicon/amplicon versions of vaccines. Replication-defective viruses have also served as important probes of the host immune response in helping to define the importance of the first round of infected cells in the host immune response, the mechanisms of activation of innate immune response, and the role of the complement pathway in humoral immune responses to viruses. PMID- 16364754 TI - New viral vaccines. AB - Vaccination is the most effective medical intervention against diseases caused by human viral pathogens. Viral vaccines prevent or modify the severity of illness in the individual and interrupt or reduce the transmission of the pathogens to other susceptible people. Through these mechanisms, vaccines against smallpox, polio, measles and hepatitis B have had an enormous impact on world health over the last 50 years. Advances in basic virology and understanding of human immunity promise more progress in the control of human viral diseases as the 21st century begins. Some important targets, including human immunodeficiency virus, respiratory syncytial virus and hepatitis C virus present challenges that require more basic research. The purpose of this review is to highlight four new viral vaccines that have recently, or will soon demonstrate the effective translation of basic investigations into clinical benefits for disease control in healthy and high-risk populations. These vaccines include the live attenuated vaccines against the RNA viruses, rotavirus and influenza A and B, and vaccines against human papilloma virus and varicella-zoster virus, which are DNA viruses that cause morbidity and mortality through their capacity to establish persistent infection. Although only the influenza vaccine has been licensed in the United States, these other new tools for disease prevention are likely to be introduced within the next few years, with profound effects on the diseases that they cause. Hence, as Virology celebrates its 50th anniversary, it is appropriate to examine these examples of recent advances in viral vaccines. PMID- 16364755 TI - How the American Society for Virology was founded. AB - The American Society for Virology, the very first such Society to be formed anywhere, was founded at a meeting of some 40 virologists at Chicago O'Hare International airport on June 9, 1981. They met after a decade and a half of intense discussion that originated at the 9th International Congress of Microbiology in Moscow in 1966 when a small group of virologists requested the International Association of Microbiological Societies to form a Virology Section within IAMS, and this request was rejected. Virologists therefore held their own First International Congress of Virology in Helsinki in 1968 which was very successful and generated intense informal discussion among leading virologists in this country as to the desirability of founding an American society for virologists. Proposals were circulated and discussed which resulted in the informal Chicago meeting that created the mechanism for founding the ASV and organizing its 1st Annual Meeting at Cornell in Ithaca in August 1982. PMID- 16364756 TI - A genome-wide study of allelic imbalance in human testicular germ cell tumors using microsatellite markers. AB - Testicular germ cell tumors (TGCT) arise by multistep carcinogenesis pathways involving selective losses and gains of chromosome material. To locate cancer genes underlying this selection, we performed a genome-wide study of allelic imbalance (AI) in 32 tumors, using 710 microsatellite markers. The highest prevalence of AI was found at 12p, in line with previous studies finding consistent gain of the region in TGCTs. High frequency of AI was also observed at chromosome arms 4p, 9q, 10p, 11q, 11p, 13q, 16q, 18p, and 22q. Within 39 candidate regions identified by mapping of smallest regions of overlap (SROs), the highest frequency of AI was at 12p11.21 approximately p11.22 (62%), 12p12.1 approximately p13.1 (53%), 12p13.1 approximately p13.2 (53%), 11q14.1 approximately q14.2 (53%), 11p13 approximately p14.3 (47%), 9q21.13 approximately q21.32 (47%), and 4p15.1 approximately p15.2 (44%). Two genes known to be involved in cancer reside in these regions, ETV6 at 12p13.2 (TEL oncogene) and WT1 at 11p13. We also found a significant association (P = 0.02) between AI at 10q21.1 approximately q22.2 and higher clinical stage. This study contributes to the ongoing search for genes involved in transformation of germ cells and provides a useful reference point to previous studies using cytogenetic techniques to map chromosome changes in TGCTs. PMID- 16364757 TI - Cytogenetic and molecular cytogenetic characterization of the stable ovarian carcinoma cell line (OvBH-1). AB - Detailed characterization and identification of cancer cell lines is the basis for the credibility of experimental studies. Therefore, chromosomal analysis should be routinely included in the protocol of cell line characterization and in the protocols of experimental studies performed on cell lines. In 2000, our group established and characterized cytomorphologically and immunophenotypically a new cell line, OvBH-1, which was derived from the ascitic fluid cells of an untreated patient with ovarian clear-cell adenocarcinoma. The aim of the current study was to characterize OvBH-1 cytogenetically and to monitor its stability by comparison of morphologic, immunohistochemical, and cytogenetic features between the early (135) and late (385) passages. Conventional and molecular cytogenetic analyses (fluorescence in situ hybridization and spectral karyotyping) of OvBH-1 revealed the following hypotriploid karyotype with random translocations: der(2)t(2;13),der(4)t(4;22), der(5)t(2;5). Complex rearrangements involving chromosomes 3, 15, and 20 were also found. FISH analysis with a p53 probe indicated the deletion of this region in two out of three copies of chromosome 17. The morphologic and immunophenotypic features, as well as the karyotypes observed in OvBH-1 in passages 135 and 385, were comparable. The monoclonality of the cell line was confirmed in a single cell cloning experiment. Our study indicated that OvBH-1 is characterized by a distinct karyotype and remains stable over 250 passages. Taking into account its thermosensitivity, its unusual karyotype, and its stability, this line can be considered as a valuable model for various experimental studies. PMID- 16364758 TI - Bell-shaped nuclei dividing by symmetrical and asymmetrical nuclear fission have qualities of stem cells in human colonic embryogenesis and carcinogenesis. AB - Large cell nuclei with at least eight distinct morphologies have been discovered throughout the fetal gut (5-7 weeks), colonic adenomas, and adenocarcinomas, five of which are not present in the normal adult colon. The most remarkable nuclear forms are hollow bells, approximately 10-15 microns in height and about 7-10 microns in bell mouth diameter. When encased in tubular syncytia, these bell shaped structures divide symmetrically by an amitotic nuclear fission process resembling the separation of two paper cups. Seven other nuclear morphotypes emerge from the bell-shaped nuclei within the syncytia by asymmetrical amitotic nuclear fission. Cells containing these differentiated nuclear forms subsequently divide extra-syncytially by mitoses that form clonal populations of cells with identical nuclear morphotypes in embryos, adenomas, adenocarcinomas, and metastases. Cells with bell-shaped nuclei thus appear to be responsible for both net growth and differentiation in the embryonic gut, adenomas, and adenocarcinomas, and fulfill the requirements for post-embryonic stem cells in colon organogenesis and carcinogenesis. PMID- 16364759 TI - Genetic heterogeneity of the MYC oncogene in advanced juvenile angiofibromas. AB - Despite their benign histological appearance, juvenile angiofibromas sometimes exhibit an aggressive growth behavior. Molecular and genetic analyses have detected beta-catenin mutations and androgen receptor gene gains in this tumor. Because intensive cross-talk among beta-catenin, androgen receptor, and C-MYC has been detected recently, we analyzed expression of the C-MYC protooncogene (MYC) on the genetic, transcriptional and translational level in seven sporadic juvenile angiofibromas. Two-color in situ hybridization analyses for chromosome 8 and MYC found in all seven juvenile angiofibromas significant MYC losses. In the three advanced juvenile angiofibromas of this series (Fisch stages III and IV) additional significant MYC gains were observed demonstrating a genetic heterogeneity for the MYC protooncogene. In cases of genetic MYC heterogeneity, reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, Western blot investigations, and immunohistology showed increased C-MYC mRNA and protein levels. Semiquantitative RT-PCR analyses from laser microdissected endothelial cells and fibroblasts found no differences of C-MYC mRNA levels, leaving open the question of the neoplastic cell in juvenile angiofibromas. The finding of genetic MYC heterogeneity associated with C-MYC overexpression on the mRNA and protein level in advanced juvenile angiofibromas indicates involvement of the MYC oncogene in aggressive growth behavior. PMID- 16364760 TI - Multitarget fluorescence in situ hybridization and melanoma antigen genes analysis in primary bladder carcinoma. AB - Conventional urine cytology has a poor prognostic performance for detecting bladder cancer, particularly for low-grade tumors. Fluorescence in situ hybridization (FISH) for chromosomes altered in bladder cancer and testing for antigens selectively expressed in tumors are promising alternatives. This study investigated the use of FISH for detecting aneuploidy of chromosomes 3, 7, 17, and 9p21 and reverse transcriptase PCR (RT-PCR) for the expression of melanoma associated antigen (MAGE) genes for the diagnosis of bladder cancer in voided urine specimens. The two techniques were compared with cystoscopic bladder biopsy results in 47 patients with urothelial cancer and 15 patients with benign prostatic hyperplasia. FISH detected cancer in 42 of 47 patients (89.4%). This was significantly higher than the detection rate 30 of 47 patients (64.3%) by MAGE RT-PCR (P < 0.001). The sensitivity of FISH increased with histologic grade and stage of the tumors, correctly identifying 77.8% of pTa and pTis, 94.1% of pT1, and 100% of Pt2-4 tumors. MAGE, however, showed a decreased sensitivity in high grade advanced tumors; it was positive in 66.7% of pTa and pTis, 70.6% of pT1, and 50% of Pt2-4 tumors. Together, the tests correctly identified urothelial cancer in 46 of 47 patients (97.9%). Combined FISH and MAGE RT-PCR testing may offer a promising alternative to conventional urine cytology in screening high risk populations and in monitoring bladder cancer patients for recurrent tumor. PMID- 16364761 TI - EGFR intragenic loss and gene amplification in astrocytic gliomas. AB - We have studied EGFR gene amplification and allelic status of chromosome 7 in 68 tumors consisting of 34 WHO grade IV glioblastomas (26 primary and 8 secondary), 14 WHO grade III anaplastic astrocytomas, and 20 WHO grade II astrocytomas, by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP), quantitative PCR, and microsatellite analysis. EGFR gene amplification was present in 27 of these tumors (40%), and we identified allelic losses at 7p11 approximately p14 in 38 of the 68 cases (56%), including 17 tumors displaying loss for EGFR intragenic markers. The positive correlation (P < 0.05, chi(2)) between tumors with EGFR intragenic loss and EGFR gene amplification, frequently displaying the EGFR vIII form, suggests that EGFR gene rearrangement leading to intragenic loss is a molecular event that participates in the amplification process of this gene. PMID- 16364762 TI - Cytogenetic abnormalities in 106 oral squamous cell carcinomas. AB - We report karyotypic features of 106 short-term cultured oral squamous cell carcinomas (SCC), 51 new and 55 previously reported cases, with clonal chromosome aberrations. The major cytogenetic findings were as follows: simple karyotypic changes were present in 38 cases (36%) and 68 tumors (64%) displayed complex karyotypes. The most common numerical changes were +7, +8, +9, +16, +18, +20, and -4, -10, -13, -14, -18, -19, -21, -22, and -Y. Structural rearrangements frequently (43% of the breaks) affected the centromeric regions, resulting in the formation of isochromosomes and whole-arm translocations. Among the recurrent structural aberrations identified, the most common were i(1q), i(3q), i(5p), i(8q), del(16)(q22), and hsr. With the exception of chromosomal band 11q13, which was involved in 25 tumors, only centromeric or near-centromeric bands were commonly involved: 3p11 approximately q11 (59 cases), 8p11 approximately q11 (57), 1p11 approximately q11 (48), 13p11 approximately q11 (46), 5p11 approximately q11 (41), 14p11 approximately q11 (41), and 15p11 approximately q11 (37). Losses of genetic material dominated over gains. The most frequent imbalances included loss of 2q33 approximately qter, 3p, 4p, 6q, 8p, 10p, 11q, 13p, 14p, and 15p, and chromosomes 18, 21, 22, and Y, and gain of chromosomes 7 and 20, 8q, and 11q13. No major karyotypic differences could be discerned between the present series of oral SCC and a previously reported series of laryngeal SCC, indicating that common genetic pathways are involved in the initiation and progression of SCC irrespective of site of origin. PMID- 16364763 TI - Previously unidentified complex cytogenetic changes found in a pediatric case of solid-pseudopapillary neoplasm of the pancreas. AB - Solid pseudopapillary neoplasm of the pancreas (SPNP) is a rare tumor with low malignant potential found in adolescent girls and young women. The pathogenesis of SPNP remains uncertain and its management is controversial. Genetic changes associated with SPNP have seldom been reported. We describe here the cytogenetic investigation of a case of SPNP in a 13-year-old girl whose tumor cells revealed two unrelated clones: one clone characterized by complex karyotypic changes, including breakpoints in two common fragile sites at chromosome 2, band q33, and chromosome 4, band q31, and the second clone defined by partial monosomy for chromosome X. Loss of heterozygosity for HRAS was also identified by array comparative genomic hybridization (a-CGH). These cumulative changes seem insufficient for activation of cell transformation, but could possibly play a role in priming the cell for future mutagenic events. PMID- 16364764 TI - Study of chromosomal abnormalities in 11 cases of cervical dysplasia using comparative genomic hybridization on cotton-lint cervical samples. AB - Comparative genomic hybridization (CGH) allows the analysis of chromosomal imbalances without requiring cell cultures and is more reliable than conventional cytogenetic studies for detecting gains, losses, and amplified regions. To perform CGH on cervical lesions, some authors obtain the tumoral DNA from frozen or paraffin-embedded biopsies. Others use laser microdissected material from paraffin-embedded samples, followed by degenerate oligonucleotide primer polymerase chain reaction (DOP-PCR). In all these cases, surgery is required to obtain the sample. In our study, we obtained DNA from a cotton-lint cervical sample obtained from the pathological zone using a colposcopy technique. Chromosomal alterations were found in 9 (81%) of the 11 cases analyzed. The most frequent alterations affected the 3p12, 4q25, 5q15 approximately q21, and 18p11 regions. Satisfactory results have been observed when the cotton-lint cervical sample has been used as the source for obtaining DNA. In the laboratory, the manipulation of this type of sample obtained by a noninvasive system is much simpler, easier, and faster than the obtained with a conventional biopsy. PMID- 16364765 TI - Lack of association between DNA repair gene ERCC1 polymorphism and risk of lung cancer in a Chinese population. AB - The ERCC1 (Excision Repair Cross Complementation Group 1) gene is involved in the nucleotide excision repair pathway. This study was designed to examine whether ERCC1 Asn118Asn (G19007A) polymorphism, which has been associated with risk of some cancers among Caucasians, may be associated with risk of lung cancer in a Chinese population. ERCC1 Asn118Asn (G19007A) genotypes were determined in DNA samples from 151 cases and 143 controls. The distribution of genotypes between cases and controls was not associated with an increased risk of lung cancer (AA versus GG: adjusted OR (odds ratio) = 1.41, 95% CI (confidence interval) = 0.76 2.59; AG versus GG: adjusted OR = 0.78, 95% CI = 0.47-1.29; and AA + AG versus GG: adjusted OR = 0.93, 95% CI = 0.73-1.19). The frequency A (0.20) of the A allele was significantly lower among these Chinese controls than in the Caucasian control populations (A = 0.54-0.65) (All P < 0.001). No statistically significant effects of age, histological subtype or smoking were found. These findings suggest that ERCC1 Asn118Asn (G19007A) polymorphism may play a limited role for lung cancer in this Chinese population. PMID- 16364766 TI - Acute megakaryoblastic leukemia and loss of the RUNX1 gene. AB - Since the RUNX1 gene contributes to megakaryopoiesis and acquired trisomy 21 is the most frequent numerical chromosome anomaly in acute megakaryoblastic leukemia (AMLK), a systematic study of RUNX1 abnormalities was performed by fluorescence in situ hybridization in AMLK patients. Four abnormalities were detected among 15 patients. One copy of RUNX1 was completeley or partially lost in three patients and translocated onto Xq24 in the fourth. The possible consequences of RUNX1 haploinsufficiency are discussed. PMID- 16364767 TI - Jumping translocation of 17q11 approximately qter and 3q25 approximately q28 duplication in a variant Philadelphia t(9;14;22)(q34;q32;q11) in a childhood chronic myelogenous leukemia. AB - The virtually obligatory presence of the Philadelphia chromosome may suggest a causal homogeneity, but chronic myelogenous leukemia (CML) is a clinically heterogeneous disease. This may be a consequence of the variable BCR breakpoints on chromosome 22 and of nonrandom secondary chromosomal abnormalities. We present the case of a boy, age 12, investigated in blastic phase of CML. Karyotyping with conventional and multiplex fluorescence in situ hybridization (FISH and M-FISH) karyotyping, complemented with reverse transcriptase-polymerase chain reaction, identified a variant Philadelphia translocation t(9;14;22)(q34;q32;q11) involving a cryptic BCR/ABL fusion with formation of the p190(Bcr-Abl) oncoprotein. M-FISH revealed also an unbalanced jumping translocation of 17q11 approximately qter alternatively present on chromosomes 14 or 20, apparently hithertofore unreported in hematological malignancies. Another secondary aberration, dup(3)(q25q28), was revealed by multipoint interphase FISH (mpI-FISH). Gain of this region is known in adult hematological malignancies and solid tumors, suggesting its general involvement in tumor initiation or progression (or both), regardless of tissue origin. PMID- 16364768 TI - Frequency of chromosomal aneuploidies and deletions of the RB and TP53 genes in MALT lymphomas harboring the t(14;18)(q32;q21). AB - The t(14;18)(q32;q21) involving the MALT1/MLT and IGH genes has been identified recently as a recurrent abnormality in mucosa-associated lymphoid tissue (MALT) lymphomas. The frequency of secondary chromosomal aberrations in MALT lymphomas harboring the t(14;18) is largely unknown. We therefore analyzed six t(14;18) positive MALT lymphomas (five parotid, one conjunctiva) by interphase fluorescence in situ hybridization for aneuploidies of chromosomes 3, 7, 12, 18, and X, gains or disruption of the CMYC/8q24 and BCL6/3q27 genes, as well as deletions of the retinoblastoma and TP53 tumor suppressor genes. Except for one MALT lymphoma of the parotid with trisomy 3, neither aneuploidies nor deletions were detected in any of our cases. PMID- 16364769 TI - Cryptic 6p21.3 duplications and triplication involving HMGA1 partially masked by add 6p in four cases of myelodysplasia. AB - Rearrangements of 6p are frequent in both myeloid and lymphoid malignant hematological disorders. High-mobility group AT-hook 2 (HMGA2) rearrangements have been described in myelofibrosis with myeloid metaplasia (MMM) and also in myelodysplasia. High-mobility group A proteins are nonhistone nuclear proteins that bind DNA and regulate the transcriptional activity of many genes. We used FISH, with bacterial artificial chromosome RP11-513I15 probe, to study 16 cases of myeloid malignancies with chromosome 6 short arm rearrangements, most of them following myeloproliferative disorders. Among these we found two 6p21.3 duplications and one 6p21.3 triplication involving HMGA1 in four cases of myelodysplasia with and without myelofibrosis. In these four cases, duplications and triplication were partially masked at the cytogenetic level by a derivative chromosome 6 resulting from translocation with another chromosome. HMGA1 proteins have been recently found overexpressed in human leukemias, but to our knowledge this is the first reported duplication of HMGA1. PMID- 16364770 TI - FISH banding in tumor cytogenetics. PMID- 16364771 TI - The translocation (4;12)(q31;q21) in myelofibrosis associated with myelodysplastic syndrome: impact of the 12q21 breakpoint. PMID- 16364772 TI - Schwannoma with a reciprocal t(9;22)(q22;q13). PMID- 16364773 TI - Trisomy 4 as sole karyotypic abnormality in acute lymphoblastic leukemia: different clinical features and treatment response between B and T phenotypes? PMID- 16364775 TI - Pain management and anesthesia in veterinary dermatology. PMID- 16364776 TI - Lasers in veterinary dermatology. PMID- 16364777 TI - Updates in therapeutics for veterinary dermatology. PMID- 16364778 TI - Skin diseases of animals in shelters: triage strategy and treatment recommendations for common diseases. PMID- 16364779 TI - Recommendations for the management and treatment of dermatophytosis in animal shelters. PMID- 16364780 TI - Feline facial dermatoses. PMID- 16364781 TI - Canine and feline eosinophilic skin diseases. PMID- 16364782 TI - Atopy: new targets and new therapies. PMID- 16364783 TI - Food allergies: update of pathogenesis, diagnoses, and management. PMID- 16364784 TI - Bacterial skin diseases: current thoughts on pathogenesis and management. PMID- 16364785 TI - German Shepherd Dog pyoderma. PMID- 16364786 TI - Updates on the management of canine epitheliotropic cutaneous T-cell lymphoma. PMID- 16364787 TI - Update on canine demodicosis. PMID- 16364788 TI - Sebaceous adenitis. PMID- 16364789 TI - Vesicular cutaneous lupus. PMID- 16364791 TI - Repeated visual and computer-assisted carotid plaque characterization in a longitudinal population-based ultrasound study: the Tromso study. AB - In a longitudinal population-based ultrasound survey, we evaluated the reproducibility of carotid plaque detection, off-line vs. online visual classification of plaque echogenicity and computer-assisted plaque echogenicity (grey-scale median, GSM) classification and plaque area measurements. The number of paired observations in the reproducibility analyses was 107 in the baseline study and 83 in the follow-up study. In addition, 198 and 222 images were selected from the baseline and the follow-up study for GSM- and plaque-area analyses. The total number of plaque images (11,160) was used to obtain comparative reference values. Despite good agreement in the reproducibility study (kappa values ranging from 0.52 to 0.57), there was a substantial drift in online visual classification of plaque echogenicity during the survey period. Inter- and intraobserver agreement on computer-assisted GSM classification was substantial, with kappa values (95% CI) of 0.77 (0.73 to 0.80) and 0.79 (0.75 to 0.84), respectively. A systematic bias in plaque area measurements was observed. Visual online classification may introduce systematic measurement errors that are not intercepted in a reproducibility study of restricted duration. Computer-assisted off-line classification had better reproducibility. However, the method is influenced by measurement errors, both in the outlining of the plaque and in the standardization procedure. PMID- 16364792 TI - Predicting fetal growth restriction with liver volume by three-dimensional ultrasound: efficacy evaluation. AB - It is well-documented that fetal growth restriction (FGR) may have increased risks of perinatal morbidity and mortality. Early detection of FGR is crucial in prenatal care and daily practice. We undertook a prospective and cross-sectional study using quantitative 3-D ultrasound (US) to assess the efficacy of fetal liver volume (LV) in predicting FGR. During the study period, 42 fetuses with FGR and 375 fetuses without FGR were included for the LV assessment in utero by 3-D US. All the fetuses were singletons and had follow-up to delivery to ensure whether they were complicated with FGR or not. Our results revealed that fetal LV assessed by 3-D US can differentiate well fetuses with FGR from those without FGR. The sensitivity of fetal LV in predicting FGR was 97.6%, with specificity 93.6%, predictive value of positive test 63%, predictive value of negative test 99.7% and accuracy 94%. In conclusion, fetal LV assessed by quantitative 3-D US can be used to predict fetuses with FGR antenatally. Our data support that fetal LV assessment by 3-D US would be a useful test in detecting fetuses with FGR. PMID- 16364793 TI - Automatic timing of aortic valve closure in apical tissue Doppler images. AB - Ultrasound color tissue Doppler imaging (TDI) can be used to estimate velocities of moving left ventricular cardiac tissue. Aortic valve closure (AVC) can be observed as a notch in apical TDI velocity/time curves occurring after ejection, but before early relaxation. This work sought to evaluate automatic and automated algorithms using TDI for timing AVC. Mitral valve position and the time point of early relaxation were extracted and used to accomplish the task. To test the algorithms, phonocardiogram of the second heart sound was recorded simultaneously with TDI and used as a reference method. The algorithms were tested on apical views of 16 healthy subjects. In 98% of the cardiac cycles, the automatic algorithm estimated the time point of AVC within 25 ms of the reference. Automatic detection of AVC might save manual effort and provide a marker separating ejection and diastole for further automated analysis. PMID- 16364794 TI - B-mode compound imaging in mice. AB - Cross-sectional B-mode images were obtained from a dead mouse for a 360 degrees scan around the mouse using a 12-MHz linear array. For each cross-section, a set of aligned images was obtained after rotation about the isocenter, which were added to produce a single compound image. The compound images demonstrated a substantial improvement over single B-mode images, with uniform image quality, low noise and improved visualization of structures. This technique may be of interest in forming the basis for a new 3-D in vivo technique in the abdomen and pelvic regions, providing high-quality ultrasound images that are not dependent on operator skill. A further development worth pursuing for improved spatial resolution is reconstruction-based tomography. PMID- 16364795 TI - Strain estimation in abdominal aortic aneurysms from 2-D ultrasound. AB - The rupture risk of abdominal aortic aneurysms (AAAs) is routinely inferred from the maximum diameter of the AAA. However, clinical experience indicates that this criterion has poor accuracy and that noninvasive assessment of the elastic properties of the vessel might give better correspondence with the rupture risk. We have developed a method for analysis of circumferential strain in AAAs from sequences of cross-sectional ultrasound B-mode images. The algorithm is fast, semiautomatic and well-suited for real-time applications. The method was developed and evaluated using data from 10 AAA patients. The preliminary results demonstrate that the method is sufficiently accurate and robust for clinically acquired data. An important finding is that local strain values may exceed the circumferential average strain significantly. Furthermore, the calculated strain shows no apparent covariation with the diagnosed diameter. This implies that the method may give new and essential information on the clinical condition of the AAA. PMID- 16364796 TI - Stationary clutter rejection in echocardiography. AB - Clutter is one of the most problematic artifacts in echocardiography. It sometimes blocks substantial portions of the image, making the diagnosis in these areas difficult, if not impossible. This is, to our knowledge, the first study aimed solely at automatic clutter rejection, performed in postprocessing, without changing the data-acquisition method. The procedure is based on the fact that the motion of the organs causing most of the clutter (e.g., the ribcage and the lungs) is much slower than that of the cardiac muscle, so that the clutter shows very small changes during a single cardiac cycle. The algorithm has been successfully tested on a set of 16 cineloops in apical two-chamber and apical four-chamber views, belonging to 16 different patients. The results show a high probability of clutter detection, while maintaining a low probability for erroneous detection of pixels as clutter. PMID- 16364797 TI - How to distinguish between ischemic and nonischemic postsystolic thickening: a strain rate imaging study. AB - Ongoing myocardial thickening after aortic valve closure (postsystolic thickening = epsilonPST) is an established marker for the presence of segmental ischemia. However, epsilonPST may also be present in late activated segments and can be induced by pharmacological interventions or left ventricular pressure overload. The aim of this study was to determine if it is possible to distinguish between ischemic and nonischemic epsilonPST. In an experimental pig-model (n = 11) regional radial deformation was measured in the inferolateral wall during either normal perfusion or regional ischemia using ultrasonic strain rate imaging. Ischemia was induced by active hypoperfusion of the circumflex coronary artery territory. Measurements were made at 1. baseline, and during 2. theodrenalin infusion, 3. dobutamine infusion 4. esmolol infusion and 5. during a preload increase induced by saline infusion. In all segments where thickening was ongoing after aortic valve closure, the amount of epsilonPST was calculated as the difference of maximal strain minus systolic strain. In addition, peak strain rate during the isovolumetric relaxation period was extracted. During normal coronary perfusion, 73% of all segments (n = 40) developed epsilonPST. This physiological epsilonPST averaged 5 +/- 2% and was most frequently induced during the esmolol infusion (n = 11). Peak isovolumetric strain rate averaged -2.1 +/- 0.5 s(-1) in segments with physiological epsilonPST. During coronary hypoperfusion, 96% of the "at risk" segments developed epsilonPST. EpsilonPST in the ischemic segments averaged 14 +/- 3%, and was highest during the dobutamine infusion (25 +/- 4%) and lowest during the esmolol infusion (5 +/- 1%). In contrast to normally perfused segments, peak isovolumetric strain rate was positive in the ischemic segments and averaged 2.0 +/- 0.5 s(-1) in these pathologic segments with postsystolic strain. Using a cut-off value of > or = 0 s(-1) for isovolumetric strain rate, pathologic epsilonPST was detected with a sensitivity of 100% and a specificity of 87%. These experimental findings were confirmed by a subsequent clinical study with 6 patients with acute myocardial infarction (ischemic group) and 6 patients with arterial hypertension or aortic stenosis (nonischemic group). Ischemic and nonischemic postsystolic thickening can be precisely differentiated by extracting the polarity of the peak isovolumetric strain curve. PMID- 16364798 TI - Analysis of contrast in images generated with transient acoustic radiation force. AB - Several mechanical imaging methods are under investigation that use focused ultrasound (US) as a source of mechanical excitation. Images are then generated of the tissue response to this localized excitation. One such method, acoustic radiation force impulse (ARFI) imaging, utilizes a single US transducer on a commercial US system to transmit brief, high-energy, focused acoustic pulses to generate radiation force in tissue and correlation-based US methods to detect the resulting tissue displacements. Local displacements reflect relative mechanical properties of tissue. The resolution of these images is comparable with that of conventional B-mode imaging. The response of tissue to focused radiation force excitation is complex and depends upon tissue geometry, forcing function geometry (i.e., region of excitation, or ROE) and tissue mechanical and acoustic properties. Finite element method (FEM) simulations using an experimentally validated model and phantom experiments have been performed using varying systems, system configurations and tissue-mimicking phantoms to determine their impact on image quality. Image quality is assessed by lesion contrast. Due to the dynamic nature of ARFI excitation, lesion contrast is temporally-dependent. Contrast of spherical inclusions is highest immediately after force cessation, decreases with time postforce and then reverses, due to shear wave interaction with internal boundaries, differences in shear modulus between lesions and background and inertial effects. In images generated immediately after force cessation, contrast does not vary with applied force, increases with lesion stiffness and increases as the ROE size decreases relative to the size of the structure being imaged. These studies indicate that improved contrast in radiation force-generated images will be achieved as ROE size decreases; however, frame rate and thermal considerations present trade-offs with small ROE size. PMID- 16364799 TI - Investigating perfluorohexane particles with high-frequency ultrasound. AB - Submicron particles filled with liquid perfluorocarbon are currently being studied as a potential ultrasound-targeted contrast agent. The objective of this study was to evaluate the scattering properties of these particles. Sets of perfluorohexane-filled particles of different average sizes (300 nm to 1000 nm) were produced with a constant total volume fraction. The attenuation coefficient was measured in the 15- to 50-MHz frequency range and was found to increase smoothly with frequency and to be independent of the amplitude and bandwidth of the transmitted pulse. The values range from 0.31 to 0.64 dB/mm at 30 MHz for mean particle size ranging from 970 to 310 nm, respectively. The backscattering spectra of the particle solutions were measured and showed no sign of nonlinear scattering. The backscattering coefficient increased with the power 3.9 +/- 0.3 of the frequency. These results confirm that liquid perfluorocarbon droplets behave as linear Rayleigh scatterers. PMID- 16364800 TI - Surfactant-stabilized contrast agent on the nanoscale for diagnostic ultrasound imaging. AB - Ultrasound contrast agents (CA) are generally micron-sized stabilized gas bubbles, injected IV. However, to penetrate beyond the vasculature and accumulate in targets such as tumors, CA must be an order of magnitude smaller. We describe a method of achieving nanometer-sized, surfactant-stabilized CA by differential centrifugation. High g force was shown to destroy bubble integrity. Optimal conditions (300 rpm for 3 min) produced an agent with a mean diameter of 450 nm, which gave 25.5 dB enhancement in vitro at a dose of 10 microL/mL, with a 13 min half-life. In vivo, the CA produced excellent power Doppler and grey-scale pulse inversion harmonic images at low acoustic power when administered. In vivo dose response curves obtained in three rabbits showed enhancement between 20 and 25 dB for dosages above 0.025 mL/kg. These results encourage further investigation of the possible diagnostic and therapeutic benefits of using nanoparticles as CA, including passive targeting and accumulation in tumors. PMID- 16364801 TI - Spatial control of gas bubbles and their effects on acoustic fields. AB - Because microbubbles can enhance therapy, such as by cavitation or by thermal means, treatment could be confined with localization of microbubbles. This spatial control can be achieved by the vaporization of liquid-filled droplets present throughout the medium in a process known as acoustic droplet vaporization (ADV). Bubbles in the form of an orthogonal plane or "wall" can thus be created and can scatter ultrasound to enhance the proximal acoustic field while shielding distal tissues. To investigate the possible effects of a preexistent bubble wall, tissue-mimicking polyacrylamide gels embedded with perfluorocarbon droplets were insonified under various conditions. The preliminary results presented in this paper show that a bubble wall can successfully cause proximal ADV at approximately half the transmitted pressures that are required without the use of a bubble wall, while also serving as a viable shield against ADV and potential damage in distal areas. The results seen here in a gel medium are promising and suggest further development in vivo is needed. PMID- 16364802 TI - Disposition of perfluorobutane in rats after intravenous injection of Sonazoid. AB - The new ultrasound contrast agent Sonazoid was injected IV in rats at doses of 0.8 and 8 muL perfluorobutane (PFB)-containing microbubbles/kg body weight. Samples were obtained from blood, liver, spleen, fat, kidney, muscle, heart, lung and brain from both males and females and the PFB gas was analyzed using validated gas chromatography mass spectrometry methods. No differences were observed between genders or doses for any of the pharmacokinetic parameters. For all tissues, the highest concentrations were observed at the first time point (i.e., 5 min postinjection) (51% of injected dose in liver; total recovery of 69%). The highest concentrations of PFB in tissue were observed in spleen > liver > lung > kidney >> other tissues. At 24 h after dosing, the total amount of PFB remaining in the tissues was 1.9%. These data fit well with the finding that after a Sonazoid dose of 8 microL microbubbles/kg to male rats, more than 50% of the injected PFB was recovered in exhaled air by 20 min after dosing. During the first 24 h after administration, more than 96% of the PFB dose was recovered in exhaled air. PMID- 16364803 TI - Pulsed cavitational ultrasound therapy for controlled tissue homogenization. AB - Methods were investigated to acoustically control the extent to which cavitation mediated tissue homogenization is responsible for lesion formation in vitro. These results may guide potential therapeutic procedures that induce damage predominantly via mechanical disruption and, thereby, avoid limitations associated with thermal ablative modalities. Porcine myocardium was insonified at 750 kHz using pulse sequences consisting of high-amplitude pulses (22 MPa Pr) interleaved with variable-amplitude "sustaining" pulses (e.g., 6.9 MPa Pr), which were intended to provide sufficient acoustic input to maintain cavitation activity between primary pulses, but to increase the spatial peak temporal average intensity (I(SPTA)) only marginally. Using modest temporal-average intensities (e.g., I(SPTA) approximately 200 W/cm2), approximately 0.5 cm3 lesions were produced consisting of homogenate that could be irrigated away to reveal smooth cavities. The prevalence of homogenate in a given lesion was sensitive to both pulse-repetition frequency and sustaining pulse amplitude, suggesting the existence of optimum acoustic parameters for producing homogenized lesions largely via mechanical perturbation. PMID- 16364804 TI - Pluronic block copolymers: novel functions in ultrasound-mediated gene transfer and against cell damage. AB - Pluronics have been investigated as vectors for drug and gene delivery in vitro and in vivo and were demonstrated to have high efficiency for gene transfer in vivo. However, they alone do not enhance gene transfer in vitro. We examined three pluronics, F127, L61 and P85, for their effects on ultrasound (US)-mediated gene transfer in three cell lines, 3T3-MDEI, C2C12 and CHO. The polymers showed differential effects on cell viability and transfection efficiency in a dose dependent manner. All the polymers were unable to facilitate gene transfer when used alone, but enhanced US-mediated gene transfer significantly at concentrations around the critical micelle concentration in the three cell lines. F127 showed no significant toxicity at any concentration and protected the cells against US-mediated damage at a high concentration. L61 decreased cell viability significantly in a dose-dependent manner, whereas P85 showed mild toxicity when its concentration was at or above 0.05%. PMID- 16364805 TI - Low-intensity ultrasound induces angiogenesis in rat hind-limb ischemia. AB - We investigated the effect of low-intensity ultrasound (US) on tissue blood flow and angiogenesis after limb ischemia in vivo. Rats underwent surgical ligation of the femoral or the iliac arteries. Half the animals were exposed to low-intensity US (0.05 W/cm2) during three consecutive sessions. At 3 weeks postsurgery, limb perfusion was assessed using laser Doppler and angiography. Immunostaining and vascular endothelial growth factor (VEGF) messenger ribonucleic acid (mRNA) expression were performed 7 d postsurgery. US irradiation significantly improved limb perfusion in both ischemic models (p = 0.04). Angiography showed increased blood vessels in the moderate ischemia (p = 0.01), but not in the severe ischemia (p = 0.19). Histology demonstrated a significantly higher number of blood vessels and proliferating cells in US-irradiated moderate and severe ischemia (p = 0.002 and p = 0.03, respectively). VEGF mRNA was significantly higher in moderate ischemia (p = 0.02). No differences in apoptotic cell death were evident in the models. Low-intensity US significantly improved tissue blood flow and angiogenesis, irrespective of the extent of the ischemia. (E-mail: ). PMID- 16364806 TI - Sonocatalytic damage of bovine serum albumin (BSA) in the presence of nanometer anatase titanium dioxide (TiO2). AB - The nanometer anatase titanium dioxide (TiO2) (a kind of crystal type of TiO2) powder was adopted as the sonocatalyst for the damage of bovine serum albumin (BSA) used as a model protein by low-power ultrasound (US) (80 kHz, 80 W). The effects of several factors on the damage of BSA molecule were reviewed by means of ultraviolet destruction and circular dichroism spectra. It was found that the BSA molecule underwent destruction of the secondary structure and loss of the alpha-helical configuration to a certain extent under ultrasonic irradiation in the presence of nanometer anatase TiO2 powder and that the damage caused by US integrated with TiO2 was more serious than those by only US or only TiO2. Furthermore, the damage degree was aggravated with the increase of TiO2, added to saturation, and then it was slowly weakened with the excessive TiO2. When a suitable amount of acid or base was added into the BSA solution, the sonocatalytic damage was also aggravated. Because the functions of proteins are decided by their space configurations, the changes of the configurations might cause the forfeiture of their function, even the apoptosis or necrosis of cells. Perhaps, an effective method of killing cancer cells by sonocatalytic damage of protein molecules in the presence of nanometer anatase TiO2 could be obtained from these experimental results. PMID- 16364807 TI - [Tissue engineering: a new space of scientific expression and technological development for health]. PMID- 16364808 TI - [Legal framework relating to human tissues used for research ends]. AB - Development in cell and tissue engineering needs human tissue samples. If French jurisdiction concerning the human tissue sample collected in a therapeutic goal is well established, the French and European legal context concerning the scientific research is not clear and controversial. In our lab, we aim to conjugate the professional and the moral duty and to impose on our researchers the respect of strictly defined procedures. In order to organize the management of these biological resources, we chose not only to take into account the present legal context concerning the collection of tissues for research purposes, but also to precede the French legal framework by inspiring from good practice, concerning on one hand the conservation, the transformation and the transport of human tissues used to therapeutic ends (decree of December 29, 1998) and on the other hand, from the ethical recommendations of the european directives. It is why, we put some procedures in place to guarantee the donor's information, the staff's security, the confidentiality as well as the tracability. PMID- 16364809 TI - Mechanobiology of bone tissue. AB - In order to obtain bones that combine a proper resistance against mechanical failure with a minimum use of material, bone mass and its architecture are continuously being adapted to the prevailing mechanical loads. It is currently believed that mechanical adaptation is governed by the osteocytes, which respond to a loading-induced flow of interstitial fluid through the lacuno-canalicular network by producing signaling molecules. An optimal bone architecture and density may thus not only be determined by the intensity and spatial distribution of mechanical stimuli, but also by the mechanoresponsiveness of osteocytes. Bone cells are highly responsive to mechanical stimuli, but the critical components in the load profile are still unclear. Whether different components such as fluid shear, tension or compression may affect cells differently is also not known. Although both tissue strain and fluid shear stress cause cell deformation, these stimuli might excite different signaling pathways related to bone growth and remodeling. In order to define new approaches for bone tissue engineering in which bioartificial organs capable of functional load bearing are created, it is important to use cells responding to the local forces within the tissue, whereby biophysical stimuli need to be optimized to ensure rapid tissue regeneration and strong tissue repair. PMID- 16364810 TI - Controlling cell biomechanics in orthopaedic tissue engineering and repair. AB - Tissue engineering offers an alternative approach with great potential for the treatment or replacement of damaged tissues or organs. In contrast to current treatments, a small sample of cells can be collected from the patient and cultured in vitro, greatly increasing the number of cells available for engineering tissue implants. As a result, engineered tissue implants limit the problems associated with patient trauma and undesirable immune response currently observed in surgical treatments practised in tissue and organ replacement. Mechano-transduction is known to play an essential role in bone tissue remodelling and repair. At physiological magnitudes, the effects of secondary messenger pathways, their components and local mediators generated as a direct result of mechanical load are known to result in an elevation of specific matrix protein mRNAs. Up-regulation of matrix protein production is paramount to tissue formation. Thus, mechano-transduction offers a method of producing bone tissue in vitro. However, successful transduction of mechanical stimuli from a substrate to cells is reliant upon a number of factors including cell-substrate adhesion, scaffold material mechanics and the activation of membrane channels, for example voltage-operated calcium channels (VOCC). Our research focuses on the optimisation of mechano-transduction pathways for successful bone tissue engineering. In this paper, we focus on the effects of cell-substrate adhesion, attenuation of VOCC activation states and biological conditioning of cell scaffold constructs utilising bioreactors in relation to mechano-transduction induced bone tissue production. The effects of these factors on successful bone tissue formation observed in increased matrix protein synthesis due to the optimisation of mechano-transduction pathways is discussed. PMID- 16364811 TI - [Cartilage tissue engineering: state-of-the-art and future approaches]. AB - Lesions of the articular cartilage have a large variety of causes among which traumatic damage, osteoarthritis and osteochondritis dissecans are the most frequent. Returning damaged cartilage in articular joints back to a functionally normal state has been a major challenge for orthopaedic surgeons. This interest results in large part because cartilage defects cannot adequately heal themselves. Current techniques used in orthopaedic practice to repair cartilage give variable and unpredictable results. Bone marrow stimulation techniques such as abrasion arthroplasty, drilling and microfracture produce mostly fibrocartilage. Autologous osteochondral transplant systems (mosaicplasty) have shown encouraging results. Autologous chondrocyte transplantation has led to a hyaline articular cartilage repair but little is known about the predictability and reliability of the procedure. The rapidly emerging field of tissue engineering promises creation of viable substitutes for failing cartilage tissue. Current tissue engineering approaches are mainly focused on the restoration of pathologically altered tissue structure based on the transplantation of cells in combination with supportive matrices and molecules. Among natural and synthetic matrices, collagen and polysaccharidic biomaterials have been extensively used with promising results. Recently, interest has switched to the use of mesenchymal stem cells instead of chondrocytes. Tissue engineering offers the possibility to treat localised cartilage lesions. Genetic engineering techniques using genetically modified chondrocytes offer also the opportunity to treat diffuse cartilage lesions occurring in osteoarthritis or inflammatory joint diseases. Electroporation is specially a reliable and inexpensive technique that shares with electrochemotherapy an ability to target the chondrocytes despite the barrier effect of the extracellular matrix without viral vectors. The authors review recent research achievements and highlight the potential clinical applications of new technologies in the treatment of patients with cartilage injuries. PMID- 16364812 TI - Cardiovascular tissue engineering: state of the art. AB - In patients requiring coronary or peripheral vascular bypass procedures, autogenous arterial or vein grafts remain as the conduit of choice even in the case of redo patients. It is in this class of redo patients that often natural tissue of suitable quality becomes unavailable; so that prosthetic material is then used. Prosthetic grafts are liable to fail due to graft occlusion caused by surface thrombogenicity and lack of elasticity. To prevent this, seeding of the graft lumen with endothelial cells has been undertaken and recent clinical studies have evidenced patency rates approaching reasonable vein grafts. Recent advances have also looked at developing a completely artificial biological graft engineered from the patient's cells with surface and viscoelastic properties similar to autogenous vessels. This review encompasses both endothelialisation of grafts and the construction of biological cardiovascular conduits. PMID- 16364813 TI - [Skin substitutes reconstructed in the laboratory: application in burn treatment]. AB - The development of skin substitutes started 25 years ago with the cultivation of keratinocytes to replace the epidermis of extensively burned patients. It is now possible to reproduce in vitro the two layers of skin, epidermis and dermis. Cultured epidermises are now usually used in burn centers dealing with the more severe patients. They are provided by hospital or private laboratories. Dermal substrates are some collagen matrices, which act in vivo as a guide for the reconstruction of a neodermis. Living dermis include living fibroblasts. Different models are now available for clinical use. Living skin equivalent is obtained by coculturing fibroblast and keratinocytes on a collagen support. Clinical essays are going on for chronic wounds. We present the different skin equivalent models and their clinical applications. PMID- 16364814 TI - [3D cellular models: a powerful access to cutaneous physiology and to innovative developments of cosmetic active compounds]. AB - Two different anti-ageing cosmetic actives based respectively, on dermal compartment regeneration and on dermoepidermal reinforcement, have been developed using tissular engineering models. Conjointly use of different innovative three dimensional models such as reconstructed dermis and skin built with human cells coming from variable aged donors allowed first, a better understanding of matrix modifications due to ageing and second, the screening of active ingredients highly targeted to reverse observed damages. The validity of such three dimensional models has been then confirmed by in vivo studies on healthy volunteers. PMID- 16364815 TI - Multiple opioid receptors on immune cells modulate intracellular signaling. AB - In 1979, Joseph Wybran reported his insights into the existence of different opioid receptor subtypes on T-cells. He observed that morphine and methionine enkephalin had different effects on human T-cell rosetting to sheep red blood cells. Since that time, a wide array of laboratories have shown that opiate alkyloids and opioid peptides exert pleiotropic effects on immune cell function. These compounds are immunomodulators, modifying immune responses to extracellular stimuli such as mitogens, antigens, and antibodies that cross-link the T-cell receptor. It has been demonstrated that cells involved in host defense and immunity express mRNA transcripts encoding the various opioid receptors originally described in neuronal tissues. Molecular imaging approaches have demonstrated the regulated expression of both delta and kappa opioid receptors, predominantly on T-cells. Moreover, atypical opiate and opioid binding sites are present on these cells. This review will consider the evidence for both classical and atypical opioid receptors and their effects on signaling within immune cells; our emphasis is the T-cell and its delta opioid receptor. PMID- 16364816 TI - Visceral pain and public speaking stress: neuroendocrine and immune cell responses in healthy subjects. AB - Whereas responses to psychological stressors are well-characterized, little is known regarding responses to painful visceral stimuli. We analyzed the emotional, cardiovascular, neuroendocrine, and cellular immune responses to painful rectal stimulation and psychological stress in healthy individuals. Eleven healthy subjects were studied in three conditions on separate days: painful rectal distension, public speaking stress, and rest. Blood was drawn for endocrinological and immunological analyses; heart rate and blood pressure were measured continuously; state anxiety was assessed with a questionnaire (STAI-S). Anxiety scores were highest in the rectal distension condition. This was evident following rectal distension (mean STAI-S scores: 44.2+/-3.5 post-distension vs. 36.6+/-3.8 post-speech, p<.05), but anxiety was also elevated at baseline (41.6+/ 3.9 vs. 32+/-3.2 recovery, p<.01). This anticipatory effect was reflected by elevated baseline cortisol (p<.05) and baseline ACTH (p<.01) levels, as well as circulating lymphocytes and lymphocyte subsets, including decreased basal CD3+CD4+ cells (p<.05) and increased CD16+CD56+ cells (p=.06) compared to rest. Both public speech and rectal distension induced cardiovascular activation, but the effect was more pronounced following rectal distension (+63.8+/-9.4 mmHg in response to distension vs. +36.4+/-6.2 mmHg in response to speech for systolic BP, p<.05). Different response patterns were also observed in the distribution of circulating leukocytes and lymphocyte subsets, including CD16+CD56+ cells (p<.05). An acute visceral pain stimulus causes profound emotional, neuroendocrine, and immune cell responses, which are markedly affected by anticipatory anxiety. These findings may have implications for conditions associated with visceral hyperalgesia. PMID- 16364817 TI - Effects of insulin-like growth factor-I on cytokine-induced sickness behavior in mice. AB - Central administration of insulin-like growth factor-I (IGF-I) attenuates sickness behavior in response to the cytokine inducer lipopolysaccharide. The present study was designed to determine the respective roles of the two main proinflammatory cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta), in these effects. Male CD1 mice were injected into the lateral ventricle (i.c.v.) of the brain with optimal amounts of either TNFalpha (50 ng) or IL-1beta (2 ng) that induce sickness behavior. Behavioral responses to IGF-I (0, .1, and 1 microg) also given i.c.v. were measured at various time intervals before and after treatment with the two proinflammatory cytokines. Mice treated with TNFalpha and IL-1beta lost body weight and displayed equivalent reductions in social exploration and instances of immobility. At the dose of .1 microg, IGF I attenuated these signs of sickness in TNFalpha-but not in IL-1beta-treated mice. At the dose of 1 microg, IGF-I attenuated IL-1beta-induced immobility and the reduction in social exploration but had no effect on loss of body weight. These findings indicate that IGF-I is more potent in attenuating sickness behavior induced by TNFalpha than that caused by IL-1beta, which is consistent with the relative specificity of the TNFalpha/IGF-I interactions in the brain. PMID- 16364818 TI - How to cope with unreliable office blood pressure measurement? PMID- 16364819 TI - Misclassification of blood pressure by usual measurement in ambulatory physician practices. AB - BACKGROUND: Standardized measurement of blood pressure (BP) is widely recommended but rarely followed in usual clinical practice. METHODS: We compared the classification of hypertension status of 107 patients referred by family physicians for ambulatory BP monitoring (ABPM) and with elevated clinic BP when assessed by usual clinical office measurement, a trained hypertension research nurse using a standardized measurement protocol, or an ambulatory BP monitor. RESULTS: Usual clinic readings resulted in higher BP readings than those obtained by the research nurse: mean (95% confidence interval [CI]), 10.8 (8.0 to 13.6)/4.9 (2.9 to 6.9) mm Hg, the daytime ambulatory BP 7.7 (5.1 to 10.3)/5.1 (3.0 to 7.1), and the 24-h ambulatory BP 12.1 (9.6 to 14.6)/8.9 (6.9 to 10.9). The interpretation of whether the patient had a hypertensive versus normotensive reading in the usual clinic setting differed in 42% of patients relative to standardized nurse readings. CONCLUSIONS: Following standardized technique is important for correct classification of the BP status of patients. Use of usual or casual technique results in higher readings than standardized or ambulatory BP readings. This study indicates that significant improvement in the assessment of BP is required for diagnosis and optimal management of hypertension. Consideration strongly needs to be given to the development of alternative methods of assessing BP in clinical practice. PMID- 16364820 TI - Morning blood pressure hyper-reactivity is an independent predictor for hypertensive cardiac hypertrophy in a community-dwelling population. AB - BACKGROUND: Morning blood pressure (BP) surge seems to be a risk factor for cardiovascular events. Although physical activity after arising significantly affects morning BP surge, it has remained unclear whether morning BP surge after controlling for physical activity (morning BP reactivity) is associated with target organ damage. METHODS: We performed ambulatory BP monitoring with simultaneous actigraphy and echocardiography in 120 community-dwelling Japanese subjects. We determined the waking time by actigraphy, and defined morning BP surge (MBPS) as the average of systolic BP during the 2 h after awakening minus the average of systolic BPs during the 1 h that included the lowest sleep BP. The ratio of MBPS/(sum of the 2-h physical activity after the arising time)0.5 was calculated as the morning BP reactivity (MBPR). RESULTS: In all the subjects studied (n = 120), MBPR was positively associated with left ventricular (LV) mass index (r = 0.30, P = .001). The MBPR had a positive association with both 24-h BP variability (SD) (r = 0.373, P < .001) and awake BP variability (r = 0.20, P < .05). The MBP hyper-reactive group (the highest quartile [Q4] of MBPR: n = 30) had significantly higher LV mass index than the nonreactive group (the other quartiles [Q1 to 3]: n = 90) (140 v 113 g/m(2), P < .001). Even after controlling for age, body mass index, gender, and 24-h systolic BP, the MBP hyper-reactive status still remained a strong predictor for LV hypertrophy. CONCLUSIONS: Exaggerated MBPS, adjusted for physical activity, is associated with cardiac hypertrophy independent of ambulatory BP level in a community-dwelling population. PMID- 16364821 TI - True resistant hypertension: is it possible to be recognized in the office? AB - BACKGROUND: True resistant hypertension (RH) is defined as uncontrolled office and ambulatory blood pressure (BP) in spite of an optimal regimen with at least three antihypertensive drugs. The aim of this study is to identify, in the office, clinical, laboratory, electrocardiographic, and echocardiographic variables associated with the occurrence of true RH. METHODS: These variables were recorded in a cross-sectional study involving 497 resistant hypertensive patients diagnosed by ambulatory BP monitoring as true RH (63.0%) or white coat RH (37.0%). Statistical analysis included bivariate and multivariate logistic regression. RESULTS: In bivariate analysis, true RH patients were younger, more frequently men, and had significantly higher office BP than white coat RH patients. They also had higher prevalence of physical inactivity, heart failure, and retinopathy, higher fasting glycemia, 24-h proteinuria and albuminuria, and lower serum potassium. In addition, these patients had higher electrocardiographic Sokolow and Cornell voltages and echocardiographic left ventricular mass index and hypertrophy. In multivariate logistic regression the variables best associated with true RH were male sex (P = .026), office systolic BP > or =180 mm Hg (P = .016), fasting glycemia > or =7.0 mmol/L (P = .042), serum potassium <4.5 mmol/L (P = .037), abnormal microalbuminuria (P < .001), adjusted Cornell voltage > or =2.6 mV (P = .002), and echocardiographic left ventricular hypertrophy (P = .009). In an alternative simpler model, proteinuria substituted microalbuminuria and echocardiographic data was excluded. Both predictive models have areas under receiver operating characteristic curve of 0.70. CONCLUSIONS: True RH can be recognized in the office in selected RH patients. We propose a simple scoring system with these variables that can be used in clinical practice. PMID- 16364822 TI - Managing resistant hypertension: not just a question of numbers. PMID- 16364823 TI - Adiponectin and arterial stiffness. AB - BACKGROUND: Adiponectin, an anti-inflammatory adipocytokine, is reduced in hypertension, diabetes, and coronary artery disease (CAD). Arterial stiffness, as aortic pulse wave velocity (PWV) in hypertension and diabetes, and as augmentation index (AIx) in CAD, is independently associated with cardiovascular mortality. We explored the relationship between adiponectin and arterial stiffness in essential hypertension. METHODS: Seventy-six untreated patients, 34 women, aged 47 +/- 1 years, mean +/- SEM with essential hypertension, had blood pressure (BP), carotid-femoral PWV, AIx plasma adiponectin, and proinflammatory cytokine C-reactive protein (CRP) measured using ELISA technique after an overnight fast. Results were analyzed using univariate and multiple logistic regression analysis. RESULTS: There was a significant positive relationship between log adiponectin and AIx (r = 0.33, P < .005) and plasma HDL-cholesterol (r = 0.40, P < .001). In contrast there were significant negative relationships with PWV (r = -0.24, P < .05), transit time (r = -0.37, P < .001), and pulse pressure amplification (r = -0.30, P < .005) in addition to waist circumference, waist-to-hip ratio, height, and weight. In a stepwise regression model, the independent predictors of AIx were heart rate, height, mean arterial pressure, age, and gender (R2= 0.69, P < .0001) with no contribution from adiponectin. However, for PWV (R2= 0.59, P < .0001) the independent determinants were mean arterial pressure, age, and adiponectin. CONCLUSIONS: These results show a divergent relationship between adiponectin and arterial stiffness, negative for PWV, and positive for wave reflection (AIx). Anthropomorphic factors, particularly height, weight, and heart rate may influence the relationship to the latter. Adiponectin is an independent predictor of aortic PWV but not of AIx. PMID- 16364825 TI - Salt intake affects the relation between hypertension and the T-786C polymorphism in the endothelial nitric oxide synthase gene. AB - BACKGROUND: Recent genetic studies have shown an association between the T-786C polymorphism in the endothelial NO synthase gene and coronary artery diseases, but any possible association with hypertension has been controversial. Other studies indicate the effect of restricting salt intake differ depending on individual salt-sensitivity, and the mixture of different sensitivity in study subjects may obscure the results. The objective of this study was to investigate the gene-environment interaction between the salt intake and this polymorphism. METHODS: We genotyped 281 healthy men after excluding 37 men on hypertensive therapy (mean age 44.8 +/- 11.9 years) for the mutation, and evaluated their daily salt intake using a validated food frequency questionnaire. RESULTS: A quartile classification of salt intake revealed that the blood pressure of subjects with the mutation was significantly higher than that of subjects without the mutation, but only in the 4th quartile (the highest intake group). A multiple logistic regression analysis also showed that the presence of this mutation increased the risk of hypertension only in the 4th quartile (adjusted odds ratio = 6.38, P = .025). CONCLUSIONS: The presence of this mutation alone does not significantly increase the risk of hypertension. However, high salt intake interacts with the mutation and leads to a significant increase in the risk of hypertension. The T-786C mutation warrants being considered a candidate for further study with the aim of tailor-made hypertension prevention. PMID- 16364824 TI - Atherosclerosis and the Glu298Asp polymorphism of the eNOS gene in white patients with end-stage renal disease. AB - BACKGROUND: We investigated whether the eNOS G/T polymorphism (Glu298Asp variant) is linked to the severity of carotid atherosclerosis and whether it is independent of asymmetric dimethylarginine (ADMA) in determining vascular damage in patients with end-stage renal disease (ESRD). METHODS: The eNOS polymorphism, ADMA, carotid intima-media thickness (IMT), and carotid artery (CCA) wall-to lumen ratio (an indicator of arterial remodeling) were determined/measured in 131 patients with ESRD. RESULTS: Both in the co-dominant and dominant model approach, IMT as well as CCA wall-to-lumen ratio were directly related to the T allele (P < or = .009) and these relationships held true in multiple linear regression analyses including ADMA and traditional and emerging risk factors. The relationship between eNOS genotypes and CCA wall-to-lumen ratio was further analyzed by a categorical approach and in a multiple logistic regression analysis, the odds ratio (OR) of increased CCA wall-to-lumen ratio was strongly associated to the T allele (codominant model: GG, OR = 1; GT, OR = 2.1; TT, OR = 8.2; P for trend = .01; dominant model: GG, OR = 1; GT and TT, OR = 2.7; P = .02). CONCLUSIONS: The T allele of eNOS gene is an independent predictor of intimal lesions and vascular remodeling and it is associated with the severity of atherosclerosis independently of ADMA. PMID- 16364826 TI - Three-dimensional echocardiographic and magnetic resonance assessment of the effect of telmisartan compared with carvedilol on left ventricular mass a multicenter, randomized, longitudinal study. AB - BACKGROUND: The hypothesis that left ventricular hypertrophy regression in hypertension relates to blood pressure (BP) control and to non-antihypertensive activity of some drugs was tested by comparing the effects of telmisartan and carvedilol on 24-h mean ambulatory BP and left ventricular mass (LVM) regression, measured using three-dimensional echocardiography (3-DECHO) and magnetic resonance imaging (MRI). METHODS: A total of 82 patients with mild-to-moderate hypertension and an optimal echocardiographic acoustic window were randomized to receive once-daily telmisartan 80 mg or carvedilol 25 mg for 44 weeks. RESULTS: Ten patients withdrew from the study because office diastolic BP remained >90 mm Hg. The 24-h mean ambulatory systolic/diastolic BP reductions were similar in both treatment groups (telmisartan, from 159.6 +/- 10.2/97.8 +/- 5.4 to 128.6 +/- 6.5/78.2 +/- 5.8 mm Hg; carvedilol, from 157.8 +/- 11.1/95.7 +/- 11.9 to 128.2 +/ 5.6/78.7 +/- 5.2 mm Hg). However, night-time and last 6-h mean BP reductions were nonsignificantly greater with telmisartan. Using 3-DE, telmisartan (P< .001) and carvedilol (P< .001) progressively reduced LVM index by 21.97 +/- 5.84 (15.7%) and 12.31 +/- 3.14 (9.1%) g/m2, respectively, at week 44. Similar magnitudes of reductions were observed using MRI (15.5% and 9.6%, respectively). Reductions in LVM index achieved with telmisartan were statistically superior to carvedilol (P< or = .001). CONCLUSIONS: The superior LVM regression with telmisartan versus carvedilol suggests telmisartan has a mechanism that may be beyond that of lowering BP in hypertensive patients. PMID- 16364827 TI - Left atrial systolic force and cardiovascular outcome. The Strong Heart Study. AB - BACKGROUND: The force needed to fill the left ventricle (LV) in late diastole (left atrial systolic force [LASF]) is abnormal in diseased hearts. The goal of this study was to determine whether LASF adds to prognostic prediction of other markers of preclinical cardiovascular disease in a population with a high prevalence of hypertension and diabetes. METHODS: Doppler echocardiography was performed on 2808 participants of the Strong Heart Study (age 59.3 +/- 8.0 years, 62.5% women, 43.0% hypertensive, 46.7% diabetic, and 54.1% obese) without valvular abnormalities or previous cardiovascular events. The LASF was estimated from mitral orifice area and mitral peak A velocity. RESULTS: The LASF was correlated with older age, higher BP, body mass index, creatinine, serum glucose, insulin levels, and heart rate. After controlling for clinical covariates, LASF was independently associated with higher LV dimensions, LV mass, stroke volume, and cardiac output (all P < .01). In Cox regression analysis, greater LASF was associated with a higher rate of CV events (HR = 1.033, 95% CI = 1.005 to 1.061; P = .021), independently of demographic characteristics, risk factors, LV geometry, and transmitral diastolic pattern. CONCLUSIONS: In a population-based sample of middle-aged and elderly adults with a high prevalence of hypertension and diabetes and without prevalent cardiovascular disease, LASF was associated with geometric changes of the heart and with increased rate of combined fatal and nonfatal cardiovascular events. PMID- 16364829 TI - Effect of BQ-123, an endothelin antagonist, on renal hemodynamics, tubular function, vasoactive hormones, and blood pressure in healthy humans: a dose response study. AB - BACKGROUND: Endothelin contributes to the maintenance of vascular tonus in both the systemic circulation and in regional vascular beds. The purpose of the study was to measure the effect of the endothelin A antagonist BQ-123 on renal hemodynamics, tubular function, vasoactive hormones, and blood pressure (BP) in healthy men. METHODS: In a randomized, placebo-controlled, double-blind dose response study of 11 healthy men we measured the effect of BQ-123 on glomerular filtrations rate (GFR), renal plasma flow (RPF), fractional excretion of sodium (FENa), lithium clearance (CLi), BP, and plasma concentrations of renin (PRC), angiotensin II (Ang II), atrial and brain natriuretic peptides (ANP, BNP), and vasopressin (AVP). BQ-123 was infused intravenously at the rate of 0.1, 0.2, and 0.3 mg/kg for 1 h, and the effects were measured before, during, and after infusion. RESULTS: The GFR and RPF were not significantly changed by BQ-123. The FENa was increased (20%, medium dose), and CLi was unchanged. Systolic BP remained constant, whereas diastolic BP decreased (-6.3%, medium dose), and pulse rate increased (7.1%, medium dose). BQ-123 increased both PRC (62%, medium dose) and Ang II (70%, medium dose). The changes in FENa, diastolic BP, pulse rate, and Ang II gradually increased up to medium dose, and in PRC up to high dose. The ANP, BNP, and AVP were practically unchanged by BQ-123. CONCLUSIONS: Infusion of an endothelin A antagonist resulted in an increase in renal sodium excretion despite a stimulation of the renin-angiotensin system and a decrease in diastolic BP. PMID- 16364830 TI - B-type natriuretic Peptide is associated with both augmentation index and left ventricular mass in diabetic patients without heart failure. AB - BACKGROUND: B-type natriuretic peptide (BNP) increases the risk of death and cardiovascular events in patients without heart failure, even at BNP values within the "normal" range. The reasons for this are unclear. METHODS: We performed two separate studies (n = 33 and n = 129) on subjects with type 2 diabetes mellitus in whom frank left ventricular systolic dysfunction had been excluded to ascertain whether a high-normal BNP could be identifying either greater augmentation of the ascending aortic pressure wave, increased left ventricular mass, or a subtly lower left ventricular ejection fraction (but within the normal range). RESULTS: Our results demonstrate that an increased augmentation index is an independent predictor of BNP levels even when BNP levels are within the normal range (P = .006 in study one and P = .007 in study two). A high-normal BNP also correlated with increased left ventricular mass and (P = .021) with a subtly lower left ventricular ejection fraction (P < .001). CONCLUSIONS: We found that high-normal BNP levels identified increased augmentation of the ascending aortic pressure wave as well as subtle left ventricular abnormalities. PMID- 16364831 TI - Warfarin therapy and systolic hypertension in men with atrial fibrillation. AB - BACKGROUND: Warfarin decreases the activity of matrix Gla-protein and causes extensive arterial calcification leading to increased systolic blood pressure (SBP) and pulse pressure (PP) in rats. We performed post hoc analyses of the Stroke Prevention in Non-Rheumatic Atrial Fibrillation (SPINAF) trial, aiming to analyze the effects of warfarin on SBP and PP. METHODS: The SPINAF trial was a randomized, double-blind, placebo-controlled trial comparing warfarin and placebo in 525 subjects with nonrheumatic atrial fibrillation. After exclusions for inadequate follow-up, 284 subjects with average treatment lasting 23.7 months were available for analysis (144 given placebo and 140 warfarin). RESULTS: Baseline SBP, diastolic blood pressure (DBP), mean arterial pressure (MAP), and PP were 132 +/- 18, 81 +/- 9, 98 +/- 11, and 50 +/- 16 mm Hg in the warfarin group, and 133 +/- 21, 80 +/- 13, 97 +/- 14, and 54 +/- 15 mm Hg in the placebo groups (all P values, NS). There was no statistically significant difference between the placebo and warfarin groups in any of the BP parameters at any time point. However, stratified analyses showed a significant increase in PP in warfarin-treated subjects with a history of hypertension (4.9 +/- 13.1 mm Hg v 0.2 +/- 11.9 mm Hg in the placebo group, P = 0.022), and those with baseline SBP > or =140 mm Hg (4.8 +/- 18.9 mm Hg v -3.4 +/- 11.2 mm Hg in the placebo group, P = .038). A similar trend was noted in diabetic subjects but it was not statistically significant. CONCLUSIONS: Based on the study results, treatment with warfarin does not result in increased BP in men with atrial fibrillation. However, it is possible that subjects at higher baseline cardiovascular risk may be more susceptible to the chronic effects of warfarin on arterial hemodynamics. PMID- 16364832 TI - Lactobacillus helveticus fermented milk lowers blood pressure in hypertensive subjects in 24-h ambulatory blood pressure measurement. AB - BACKGROUND: The present study was carried out to evaluate the blood pressure (BP) lowering effect and the safety aspects of Lactobacillus helveticus LBK-16H fermented milk with high tripeptide doses on hypertensive subjects using 24-h ambulatory measurements (ABPM). METHODS: In a randomized, double blinded placebo controlled parallel group study, 94 hypertensive patients not receiving any drug treatment were given 150 mL twice daily of either L. helveticus LBK-16H fermented milk with a high concentration of tripeptides (Ile-Pro-Pro 7.5 mg/100 g and Val Pro-Pro 10 mg/100 g) or a control product, for 10 weeks after a 4-week run-in period. Twenty-four-hour ABPM were taken at the beginning and at the end of the intervention period. The average baseline systolic and diastolic BP values were 132.6 +/- 9.9/83.0 +/- 8.0 mm Hg in the L. helveticus group and 130.3 +/- 9.6 /80.2 +/- 7.0 mm Hg in the control group. RESULTS: There was a mean difference of -4.1 +/- 0.9 mm Hg in systolic (P = .001) and a -1.8 +/- 0.7 mm Hg in diastolic BP (P = .048) between the L. helveticus group and the control group. There was no difference in the sum of the adverse events (P = .820). CONCLUSIONS: Lactobacillus helveticus LBK-16H fermented milk containing bioactive peptides, in daily use, does have a BP-lowering effect in hypertensive subjects and is thus a potential for the dietary treatment of hypertension. PMID- 16364833 TI - Triple vasopeptidase inhibition normalizes blood pressure in conscious, unrestrained, and spontaneously hypertensive rats. AB - BACKGROUND: CGS 35601 is a potent triple vasopeptidase inhibitor (VPI) of angiotensin-converting enzyme (ACE), neutral endopeptidase (NEP), and endothelin converting enzyme (ECE). The aim of the study was to determine the effects of this VPI on the hemodynamic profile of conscious, instrumented, unrestrained spontaneously hypertensive rats (SHR), in comparison to selective inhibitors of ACE and ACE + NEP, than +ECE combined. Circulating plasma concentrations of vasoactive mediators and reactive oxygen species were measured. METHODS: Old SHR male were instrumented (arterial catheter) and placed in a metabolic cage for daily hemodynamic measurements and blood samplings. Seven days after surgery, SHR received 1) saline vehicle; 2) increasing doses of the triple CGS 35601 (0.01, 0.1, 1, and 5 mg/kg/d, intra-arterially (i.a.) infusion for 5 d/dose) followed by a 5-day washout period; 3) benazepril (ACE inhibitor), ACE inhibitor + CGS 24592 (NEP inhibitor) and ACE inhibitor + NEP inhibitor + CGS 35066 (ECE inhibitor) (1 or 5 mg/kg/d i.a. infusion for 5 d/combination) followed by a 5-day washout period. RESULTS: The lowest dose of CGS 35601 had no effect. Doses at 0.1, 1, and 5 mg/kg/d reduced mean arterial blood pressure by 10%, 22%, and 40%, respectively. Heart rate was unaffected in all groups. CGS 35601 decreased concentrations of angiotensin II (Ang II), endothelin-1 (ET-1), and pro-atrial natriuretic peptide (proANP), and increased those of big ET-1, atrial natriuretic peptide (ANP), bradykinin (BK), and hydrogen peroxide (H2O2) dose dependently. CONCLUSIONS: The blood pressure-lowering effect of this triple VPI was superior to that of the other VPI in this preclinical rat model of hypertension. Further experiments are needed to assess triple VPI to other combinations in other models with regard to efficacy and angioedema. Only then it may constitute a first-in class approach for the treatment of hypertension and other cardiovascular disorders. PMID- 16364834 TI - Prolonged melatonin administration decreases nocturnal blood pressure in women. AB - BACKGROUND: The nocturnal decline of blood pressure (BP) is almost coincident with the elevation of melatonin, which may exert vasodilatating and hypotensive effects. In this study we investigated whether prolonged nocturnal administration of melatonin could influence the daily rhythm of BP in women. METHODS: In a randomized double-blind study, 18 women, 47 to 63 years of age and with normal BP (N = 9) or treated essential hypertension (N = 9), received a 3-week course of a slow-release melatonin pill (3 mg) or placebo 1 h before going to bed. They were then crossed over to the other treatment for another 3 weeks. In each woman ambulatory BP was recorded for 41 h at baseline at the end of each treatment period. RESULTS: In comparison with placebo, melatonin administration did not influence diurnal BP but did significantly decrease nocturnal systolic (-3.77 +/- 1.7 mm Hg, P = .0423), diastolic (-3.63 +/- 1.3 mm Hg, P = .0153), and mean ( 3.71 +/- 1.3 mm Hg, P = .013) BP without modifying heart rate. The effect was inversely related to the day-night difference in BP. CONCLUSION: These data indicate that prolonged administration of melatonin may improve the day-night rhythm of BP, particularly in women with a blunted nocturnal decline. PMID- 16364835 TI - Potential use of melatonin as adjunct antihypertensive therapy. PMID- 16364836 TI - Effects of different classes of antihypertensive drugs on cerebral hemodynamics in elderly hypertensive patients. AB - BACKGROUND: This study was designed to estimate the effect of different classes of antihypertensive drugs on cerebral hemodynamics in elderly stroke-free, nondiabetic hypertensive patients by applying frequency-domain techniques. METHODS: A total of 60 hypertensive patients divided into treatment groups of 15 unmedicated (HT), 15 nifedepine sustained release form (CCB), 15 atenolol (BB), and 15 valsartan (AIIA), and 15 age-matched healthy volunteers (CON) were studied prospectively. Variability of arterial blood pressure (ABP) and middle cerebral artery flow velocity (MCAFV) detected by transcranial Doppler sonography were diffracted into very-low frequency (VLF, 0.016 to 0.04 Hz), low frequency (LF, 0.04 to 0.15 Hz), and high frequency (HF, 0.15 to 0.4 Hz). Cerebral hemodynamics was quantified by the ABP-MCAFV transfer function. RESULTS: The ABP and MCAFV were statistically different between CON and HT groups, but not significantly different among CON, CCB, BB, and AIIA groups. The LF phase and HF magnitude in the HT, CCB, and BB groups were significantly attenuated than in those of the CON group, but not statistically different between the AIIA and CON groups. There was no statistical difference in VLF and LF transfer magnitude among the five groups. CONCLUSIONS: Although cerebral vasomotor reserve and cerebral blood flow are impaired in unmedicated hypertensive patients, the cerebral autoregulatory response was preserved both in controlled and uncontrolled hypertension. Nifedipine, atenolol, and valsartan are all effective as monotherapy to control ABP and restore cerebral blood flow. Valsartan has the effect to normalize the changes in cerebral vasomotor reserve to a level similar as in age-matched healthy subjects. PMID- 16364837 TI - Pioglitazone-induced insulin sensitization improves vascular endothelial function in nondiabetic patients with essential hypertension. AB - BACKGROUND: It has been suggested that insulin resistance is involved in the impaired vascular endothelial function not only in diabetic patients but also in hypertensive patients. The present study assessed the hypothesis that primary treatment of insulin resistance may reverse endothelial dysfunction in hypertensive subjects. METHODS: Fifteen nondiabetic patients with essential hypertension were enrolled in this study. An oral glucose tolerance test (OGTT) and insulin sensitivity test were performed. Insulin sensitivity was determined with the steady-state plasma glucose (SSPG) method. Only subjects with insulin resistance (SSPG > or =8.3 mmol/L) were included. Endothelium-dependent vasodilation during reactive hyperemia (flow-mediated dilation) was evaluated using ultrasonography. Six months after treatment with the insulin-sensitizing agent pioglitazone (30 mg/day), these examinations were repeated in all subjects. RESULTS: Pioglitazone did not decrease fasting plasma glucose and hemoglobin A1c (HbA1c) in the nondiabetic subjects, although the area under the curve for glucose and insulin on OGTT significantly decreased. A marked decrease in SSPG was observed after pioglitazone treatment (10.7 +/- 1.4 to 7.9 +/- 2.1 mmol/L, P < .001). Endothelium-dependent vasodilation evaluated by flow-mediated dilation was also improved by pioglitazone (5.0% +/- 2.2% to 6.3% +/- 2.4%, P = .023). Furthermore, the increase in flow-mediated dilation was closely correlated with the decrease in SSPG (r = 0.72, P = .002) but not with the decrease in area under the curve for glucose or insulin on OGTT. Endothelium-independent dilation induced by glyceryl trinitrate was not altered by pioglitazone. CONCLUSIONS: The present findings demonstrate that pioglitazone improves endothelial function in nondiabetic hypertensive individuals with insulin resistance, and that the improvement is associated with the amelioration of insulin resistance itself rather than that of hyperglycemia or hyperinsulinemia. PMID- 16364838 TI - Does aldosterone-to-renin ratio predict the antihypertensive effect of the aldosterone antagonist spironolactone? AB - BACKGROUND: The recognition that some 10% to 15% of the hypertensive population may have aldosterone excess has increased the frequency of measurement of the aldosterone-to-renin ratio (ARR) and the use of aldosterone antagonists. Whether this ratio will predict the blood pressure (BP) response to spironolactone is not clear. METHODS: We correlated the BP response to spironolactone 50 mg/day to baseline ARR in 69 hypertensive patients (mean [+/-SD] age 57 +/- 2 years, 65% male), consisting of 39 subjects with long-standing hypertension (4.0 +/- 0.2 years) whose hypertension was uncontrolled on at least three antihypertensive medications and 30 previously untreated patients who were randomized in a cross over design to receive either spironolactone 50 mg/day or bendroflumethiazide 2.5 mg/day for 4 weeks. RESULTS: After 4 weeks of spironolactone, BP in patients with never-treated hypertension was reduced by 18 +/- 3 / 11 +/- 1 mm Hg. There was a highly significant correlation between log ARR and the fall in systolic BP (r = 0.69, P < .001) and diastolic BP (r = 0.45, P < .05). Nine of ten patients with low renin activity (< or =0.5 ng/mL/h) showed a >20-mm Hg fall in systolic BP. No such correlations were seen when BP was reduced by bendroflumethazide 2.5 mg. For patients with resistant hypertension, despite a BP reduction of 28 +/- 3 / 13 +/- 2 mm Hg after 14 weeks of spironolactone, there was no relationship between the reduction in BP and the ARR; however, subjects with pretreatment potassium <4.0 mmol/L had a greater response than those with levels > or =4.0 mmol/L (34 +/- 3 / 16 +/- 2 v 20 +/- 6 / 8 +/- 3 mm Hg, P < .05) CONCLUSIONS: Based on the study results, ARR and low renin activity may predict the response to spironolactone in never-treated hypertensive patients but not in patients taking antihypertensive drugs, possibly because of the effect of these agents on ARR. In such patients a trial of spironolactone is required to assess the BP response. PMID- 16364839 TI - Angiotensin receptor blockers and magnesium status in essential hypertension. PMID- 16364841 TI - Mutations of the epidermal growth factor receptor in non-small cell lung cancer - search and destroy. AB - The targeting of the ATP binding pocket of the epidermal growth factor receptor (EGFR) tyrosine kinase, by the small molecule drugs gefitinib and erlotinib, represents a promising new therapeutic strategy in non-small cell lung cancer. However, it is now apparent that only a subset of patients responds to such treatment. Two publications in early 2004 reported the presence of activating mutations in the EGFR tyrosine kinase gene conferring exquisite sensitivity to these drugs. Several publications have since reported prospective data consistent with this finding. This brief review summarises the mutation data from 15 such studies in terms of mutation frequency by clinicopathological features and correlation with response to tyrosine kinase inhibition. A new paradigm for the routine detection of such mutations is needed to facilitate patient selection for treatment and further studies. PMID- 16364842 TI - Of mice and men: mouse models and the molecular mechanisms of post-transplant coronary artery disease. PMID- 16364843 TI - Utility of histologic parameters in screening for antibody-mediated rejection of the cardiac allograft: a study of 3,170 biopsies. AB - BACKGROUND: Diagnostic criteria for antibody-mediated rejection (AMR) of the cardiac allograft have recently been proposed as part of the International Society for Heart and Lung Transplantation (ISHLT) biopsy grading scheme. Histologic features of vascular adherence of macrophages (VASC) and endothelial activation or swelling in capillaries (ENDO) are proposed as criteria to prompt the immunohistochemical investigation of biopsies for AMR. The aim of this study was to determine whether VASC and ENDO are adequate to act as screening parameters to trigger further AMR investigation. METHODS: We examined our database of biopsy findings where histologic vascular parameters as well as immunofluorescence (IF) to detect AMR were collected (n = 3,170). Histologic parameters were graded semi-quantitatively on a scale from 1 to 5, where 1 = absence and 5 = obvious and generalized presence of the finding. RESULTS: Seven hundred sixty-eight of 3,170 biopsies had IF findings diagnostic of AMR in the absence of cellular rejection (ISHLT = 0). ENDO had a sensitivity of 63% and a specificity of 80%. VASC had a sensitivity of 30% and specificity of 99%. Combining the interpretation of the 2 tests did not result in a significant improvement of test sensitivity. CONCLUSIONS: Neither ENDO, VASC nor the combination of the tests indicated sufficiently high sensitivity to serve as a screening tool before further diagnostic investigation for AMR. Immunohistochemical testing remains necessary in the majority of cases to identify AMR. PMID- 16364844 TI - Changes in myocardial vasoreactivity after drastic reduction of plasma fibrinogen and cholesterol: a clinical study in long-term heart transplant survivors using positron emission tomography. AB - BACKGROUND: Given the central importance of the microvasculature in heart transplant recipients, we investigated the possibility of increasing cardiac perfusion after reduction of low-density lipoprotein (LDL)-cholesterol, lipoprotein (a), C-reactive protein (CRP) and fibrinogen plasma levels after apheresis treatment in transplanted patients. METHODS: Ten long-term heart transplant recipients were examined with positron emission tomography (PET) to measure myocardial perfusion before and after a single heparin-mediated extracorporeal LDL/fibrinogen precipitation (HELP)-apheresis treatment. PET studies were performed the mornings before and after the apheresis treatment. Myocardial blood flow at rest and during adenosine-induced hyperemia was measured using (13)N-ammonia. RESULTS: HELP-apheresis reduced the plasma levels of LDL cholesterol, lipoprotein (a) and C-reactive protein by 48% (p < 0.001), fibrinogen by 42% (p = 0.02), plasma viscosity by 14% (p = 0.004) and erythrocyte aggregation by 28% (p < 0.02). Osmolality (<1%) and hematocrit (<2%) remained stable. A single apheresis treatment increased median corrected rest flow by 17.5% (p = 0.007) and median hyperemic flow by 27% (p = 0.02). Median coronary flow reserve increased by 8.1% (p = 0.09). Hyperemic flow after adenosine infusion increased plasma vascular endothelial growth factor levels only before HELP-apheresis (+60%), indicating better ischemic tolerance after apheresis (p = 0.01). CONCLUSIONS: Myocardial perfusion in transplanted hearts increases significantly after single HELP-apheresis treatment. The present study is only a proof of concept, providing complementary evidence to clinical long-term studies showing that cholesterol reduction either with statins and/or apheresis improves heart transplant outcome. PMID- 16364845 TI - Normalization of right ventricular performance and remodeling evaluated by magnetic resonance imaging at late follow-up of heart transplantation: relationship between function, exercise capacity and pulmonary vascular resistance. AB - BACKGROUND: Right ventricular (RV) dysfunction remains one of the most prominent complications during the period immediately after heart transplantation (HT); however, late adaptation of the RV has not been well described. The aim of our study was to evaluate RV function and remodeling using magnetic resonance imaging (MRI) and to correlate it with exercise capacity and also with hemodynamic data obtained before HT. METHODS: We prospectively evaluated RV function of 25 heart transplanted patients, without cardiac allograft vasculopathy, who were documented by negative dobutamine stress echocardiography during late follow-up (Group 1, 6 +/- 4.3 years) using MRI. We then compared Group 1 with a control group consisting of 10 patients, who were < or =1 year post-HT (Group 2), hemodynamically stable, and with the same pre-operative hemodynamic features as Group 1. Their pulmonary arterial systolic blood pressure (PSBP) varied from 17 to 67 mm Hg (43.2 +/- 15.3) and pulmonary vascular resistance (PVR) from 1.0 to 5.4 Wood units (2.5 +/- 1.12). The following parameters were studied: RV end diastolic volume (EDV) and systolic volume (ESV); stroke volume (SV); ejection fraction (EF); and mass (M). We also evaluated the VO2 peak and slope VE/VCO2 values during a treadmill test. Data were analyzed and correlated with the hemodynamic values of PVR and PSBP obtained pre-HT. RESULTS: In Group 1, treadmill evaluation data showed exercise VO2 peak (19.9 +/- 3.19 ml/kg/min) and slope VE/VCO2 (36.9 +/- 4.5) values comparable to those of sedentary individuals; RV variables according to MRI were within normal ranges, with the following mean values for Groups 1 and 2, respectively: RVEDV, 99.6 +/- 4.0 ml vs 127 +/- 16 ml (p = 0.03); RVESV, 42 +/- 2 ml vs 58.5 +/- 9 ml (p = 0.01); RVSV, 57 +/- 3 ml vs 71 +/- 10 ml (p = 0.1); RVEF, 58 +/- 1.4% vs 54 +/- 3.8% (p = 0.29); and RVM, 43.4 +/- 1.9 g vs 74 +/- 8.8 g (p = 0.001). There was no correlation between hemodynamic pulmonary values before HT or any other index of late RV performance, including RV remodeling and hypertrophy, in our study population (p = not significant). CONCLUSIONS: In contrast to what we would expect for heart transplant patients at late follow-up, the RV may adapt to pulmonary pressure and resistance, with reverse remodeling characterized by volume and mass reduction, leading to normalization of RV function despite abnormal hemodynamic pulmonary values being measured before HT. There was no influence on the low exercise capacity observed in these patients, in the absence of cardiac allograft vasculopathy. PMID- 16364846 TI - Early institution of mechanical support improves outcomes in primary cardiac allograft failure. AB - BACKGROUND: Primary graft failure (PGF) is the leading cause of early mortality after cardiac transplantation, accounting for 27.1% of deaths within 30 days. PGF is defined as severe dysfunction of the cardiac allograft without any obvious anatomic or immunological cause. The purpose of this study was to analyze our last 9 years of experience with cardiac transplantation to determine predictors of PGF and the influence on survival of our policy of early institution of mechanical circulatory support (MCS) in these patients. METHODS: Data on 214 consecutive cardiac transplants performed at The Alfred Hospital between January 1996 and August 2004 were reviewed. PGF was defined as right or left or biventricular failure manifesting as hypotension (systolic blood pressure <90 mm Hg), low cardiac output (cardiac index <2.0 liter/min/m2 and pulmonary capillary wedge pressure >20 mm Hg after coming off cardiopulmonary bypass despite inotropic support of up to 5 mug/min adrenaline and without any other obvious cause for the graft dysfunction. RESULTS: PGF developed in 51 patients (24%). Significant factors in the development of PGF were long ischemic time, which became significant over 4 hours (odds ratio, 1.43; p = 0.01) and increased donor age (odds ratio, 1.027; p = 0.045). Fifteen patients required mechanical support, and of these, 10 survived to leave hospital. CONCLUSIONS: PGF is the major cause of early mortality after cardiac transplantation. Significant risks for PGF are long allograft ischemic time and increased donor age. Once the patient has survived 30 days, however, the longer-term survival is not influenced by PGF. Our management strategy of early mechanical support has yielded good outcomes in this population with a high risk of early death. PMID- 16364847 TI - Routine computed tomography screening of the chest in high-risk cardiac transplant recipients may improve survival. AB - BACKGROUND: Multiple studies have demonstrated an increased incidence of lung cancer in the heart transplant population. We reviewed our cardiac transplantation experience with respect to the development of bronchogenic carcinoma and explored the role of routine chest computed tomography (CT) in its surveillance. METHODS: We performed a review of our cardiac transplantation experience, highlighting the incidence of lung cancer, and we analyzed our recent experience with screening chest CT in lung cancer surveillance in this patient group. RESULTS: Eighteen patients developed 20 cases of bronchogenic carcinoma for an incidence of 6.83%. In 10 cases, the patients underwent surgical resection; however, in the remaining cases, the patients were either treated with chemotherapy and/or radiation or they died before initiation of therapy. The actuarial 1-, 2- and 5-year overall survival rates were 49%, 29% and 13%, respectively. The median survival of patients who underwent surgical resection was 28 months (3 to 85 months), whereas the median survival of patients who were either ineligible for surgery or died before initiation of treatment was only 1 month (1 to 13 months). All patients diagnosed with lung cancer by chest CT underwent surgical resection; however, only 37.5% of patients diagnosed with lung cancer by chest X-ray were found at an appropriate stage for resection (p = 0.025). CONCLUSIONS: Cardiac transplant recipients have a significant risk of developing bronchogenic carcinoma. Routine chest CT screening in high-risk patients may enable clinicians to identify disease earlier, which is essential for the option of surgical resection and, therefore, prolonged survival. PMID- 16364848 TI - Effect of diabetes on short- and long-term outcomes after left ventricular assist device implantation. AB - BACKGROUND: Diabetes mellitus (DM) is a frequent co-morbidity in patients with congestive heart failure (CHF). Implantation of LVADs is an acceptable option for diabetic patients with end-stage heart failure, yet no previous study has specifically examined the clinical outcomes of this patient population. METHODS: A retrospective analysis was performed on all patients who underwent LVAD insertion at a single institution from June 17, 1996 to April 14, 2004. Patients were divided into 2 groups: diabetics (DM) and non-diabetics (NDM). The groups were compared with regard to demographics, etiology of heart failure, body mass index (BMI), intensive care unit (ICU) stay, early mortality (< or =30 days), bridge-to-transplantation rate and post-LVAD and post-transplant survival. RESULTS: Two hundred one patients were identified. Of these, 49 (24.4%) had DM. Compared with the NDM group, DM patients had a higher mean body mass index (30.1 +/- 6.0 vs 26.1 +/- 4.8, p < 0.001) and a higher proportion of hypertension (57.4% vs 19.7%, p < 0.001). Although post-LVAD survival was similar, post transplant survival in DM patients was significantly lower compared with NDM patients, with 1-, 3-, 5- and 7-year actuarial survival rates of 86.9%, 71.0%, 56.5% and 56.5% vs 90.5%, 88.4%, 83.0% and 80.7% (p = 0.020), respectively. CONCLUSIONS: Carefully selected diabetic patients can be successfully bridged to transplantation using LVADs with comparable outcomes to non-diabetic patients. Reasons for diminished post-transplant survival in this cohort are unclear and necessitate the conductance of multi-institutional studies to evaluate outcomes. PMID- 16364849 TI - Impact of left ventricular assist device (LVAD)-mediated humoral sensitization on post-transplant outcomes. AB - BACKGROUND: Humoral sensitization, defined as a panel-reactive antibody (PRA) screen of >10%, places heart transplant recipients at a greater risk of acute rejection and mortality. Previous studies have suggested an increased sensitization in left ventricular assist device (LVAD) recipients, although neither the impact of device selection nor the clinical importance of elevated PRA in these patients has been completely described. METHODS: Using the registry of the International Society for Heart and Lung Transplantation (ISHLT), we compared PRA levels in 7,686 heart transplant recipients to determine the impact of LVAD therapy on humoral sensitization, acute rejection and mortality. To determine the impact of device selection on sensitization, we compared data from the ISHLT registry as well as from our own institution. RESULTS: Elevated PRA levels were found in 16.6% of LVAD recipients, compared with 7.6% of non-LVAD controls (p < 0.0001). Sensitization differed by device type, being present in 21.9% of Thoratec recipients, 14.4% of HeartMate recipients, and 15.5% of Novacor recipients (p = 0.01). Despite these findings, LVAD use had no impact on rejection rates. LVAD use was associated with a small increase (4.4% and 4.3%, respectively) in 1- and 2-year mortality. CONCLUSIONS: These findings support the concept that mechanical circulatory support increases the rate of humoral sensitization. However, these differences in sensitization do not translate to substantial differences in the clinical outcomes of rejection and mortality. PMID- 16364850 TI - Assessment of recurrent heart failure associated with left ventricular assist device dysfunction. AB - BACKGROUND: Patients with advanced heart failure may require long-term support with an intracorporeal left ventricular assist device (LVAD) before cardiac transplant, while awaiting myocardial recovery, or during destination therapy. Compared with the diagnosis of native heart dysfunction, there is less experience with the assessment of recurrent heart failure after LVAD placement. METHODS: Ten patients (9 men, 1 woman; age, 58 +/- 11 years) were studied after LVAD placement. Six patients were studied because of recurrent heart failure; the remaining 4 had other indications for study and are reported here as controls. Cardiac catheterization, including LVAD and cannulae catheterization, and angiography were performed. RESULTS: Inflow cannula valve regurgitation by LVAD angiography was found in 3 cases. Patients with regurgitation had a mean increased resting LVAD rate of 105 beats/min (range, 90-120); LVAD output exceeded forward cardiac output (LVAD - thermodilution cardiac output = +3.7 liters/min [0.6-6.4]). Inflow cannula obstruction identified with a filling phase pressure gradient between the left ventricle and the LVAD was found in 3 additional patients. Patients with obstruction had decreased resting LVAD rates (50 beats/min, all patients); LVAD output was less than the forward cardiac output (LVAD - thermodilution cardiac output = -2.3 liters/min [-0.8 to -3.5]). Compared with those with inflow valve regurgitation, patients with cannula obstruction had higher pulmonary capillary wedge pressures; phasic left ventricular pressure variation was reduced. Patients with cannula dysfunction underwent surgical intervention, and 4 of 6 were long-term survivors. CONCLUSIONS: When heart failure recurs after LVAD placement, abnormalities of the inflow cannula are common. Cardiac catheterization can confirm the diagnosis before surgical intervention. Hemodynamic coupling between the left ventricle and the LVAD is increased with inflow valve regurgitation and reduced with cannula obstruction. PMID- 16364851 TI - Microarray-based gene expression profiling of retransplanted rat cardiac allografts that develop cardiac allograft vasculopathy. AB - We studied the expression of 9,906 genes in retransplanted rat cardiac allografts that developed cardiac allograft vasculopathy (CAV) with the use of DNA microarray and real-time reverse transcriptase-polymerase chain reaction. Although only a slight difference in the timing of the retransplantation induced the later development of CAV, 1,067 genes were differentially expressed in the allografts 1 day after retransplantation. Thus, the development of CAV was determined by a robust difference in gene expression soon after retransplantation, controlled by a slight difference in retransplantation timing. In contrast, only 26 genes showed significant upregulation in the later phase of CAV development, and the time-course of the induction of 16 genes was associated with CAV progression. Of these genes, 8 were induced in 2 different aortic allograft combinations, and the time-course of the induction was correlated with the development of transplant vasculopathy. Microarray-based gene expression profiling has the potential to elucidate the mechanism of experimental chronic cardiac allograft rejection. PMID- 16364852 TI - Elevated donor troponin levels are associated with a lower frequency of allograft vasculopathy. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV) is considered a major cause of morbidity and mortality in transplant recipients and may reflect immune-mediated endothelial injury in response to the donor heart. Elevated troponin levels in the donor serum might provide a marker for this phenomenon; therefore, we evaluated the relationship of donor troponin levels to the development of CAV. METHODS: A retrospective analysis of troponin levels was undertaken from cardiac donor patients, and transplant recipients were monitored for the development of vasculopathy by angiography (N = 171). RESULTS: Angiographically significant CAV developed in 6% of transplantation patients and troponin levels were inversely related to the severity of CAV. CONCLUSIONS: Elevated donor troponin levels are not associated with the development of CAV but rather with a significantly reduced long-term risk of developing CAV, suggesting a possible protective effect of donor released protein. PMID- 16364853 TI - Activation of mitogen-activated protein kinases during human lung transplantation. AB - BACKGROUND: Ischemia-reperfusion is one of the unavoidable steps in lung transplantation; it is associated with acute inflammatory responses and cell death. The intracellular signal transduction mechanisms of these events are largely unknown. We hypothesize that activation of mitogen-activated protein kinases (MAPKs) is one of the important signaling events during human lung transplantation. METHODS: Lung tissue biopsies were performed on 15 patients undergoing transplantation: after cold ischemic preservation; after warm ischemia (implantation); and after 1- or 2-hour reperfusion. The phosphorylation status of MAPK isoforms (ERK, p38-MAPK and JNK) was examined by Western blotting. RESULTS: Phosphorylation of ERK was dramatically increased during the first 2 hours of reperfusion. Phosphorylation of JNK also significantly increased at lower levels. In contrast, phosphorylation of p38 showed no significant changes. CONCLUSIONS: We speculate that the rapid and sustained activation of ERK and JNK during the early reperfusion period may contribute to acute inflammatory responses and cell death of lung grafts. PMID- 16364854 TI - Pleural space problems after living lobar transplantation. AB - BACKGROUND: We reviewed our experience with adult living lobar lung transplant (LL) recipients to assess whether size and shape mismatch of the donor organ to the recipient pre-disposes to the development of pleural space problems (PSP). METHODS: Eighty-seven LL were performed on 84 adult recipients from 1993 through 2003. Seventy-six patients had cystic fibrosis. Patient records were examined for PSP, defined as air leak or bronchopleural fistula for more than 7 days; pneumothorax, loculated pleural effusions, or empyema in 68 patients for which complete data were available. RESULTS: There were 24 PSP identified for an overall incidence of 35%. The most common PSP was air leak/bronchopleural fistula, accounting for 38% of PSP. The second most common PSP was loculated pleural effusion (21% of PSP). Empyema was uncommon (2 patients, 3% of total patients) in our series of patients despite the large population of cystic fibrosis patients. In 4 of these patients, computed tomography-guided drainage was used for loculated effusions after chest tube removal. Three LL patients underwent surgery for persistent air leak and required muscle flap repair. One of these required subsequent omental transfer. Two LL patients required decortication for empyema. Many patients with PSP could be managed without further surgical intervention (14/24 patients). Donor-recipient height mismatch was not significantly different between PSP and non-PSP patients (p = 0.53). CONCLUSIONS: The incidence of PSP in LL recipients is similar to that reported in the literature on cadaveric transplant recipients. The relatively small lobe in the potentially contaminated chest cavity of cystic fibrosis recipients does not significantly pre-dispose to development of empyema despite immunosuppression. Many PSP can be managed non-operatively, although early aggressive intervention for large air leaks and judicious chest tube management are essential for a good outcome. PMID- 16364855 TI - Pulmonary capillaritis in lung transplant recipients: treatment and effect on allograft function. AB - The clinical outcomes of lung transplant recipients presenting with post transplant pulmonary capillaritis have not been well described. We retrospectively reviewed 40 cases of biopsy-proven pulmonary capillaritis in lung transplant recipients. Patients presented with a clinical syndrome characterized by dyspnea, hypoxemia, abnormal chest X-ray, and a decrease in forced expiratory volume in 1 second (FEV1); 25% presented with hemoptysis, and 18% with fulminant respiratory failure. Therapy with intravenous corticosteroids resulted in clinical improvement in 17 cases (43%). A response to plasmapheresis was seen in 12 (67%) of 18 cases refractory to corticosteroids. There were 5 deaths within 3 months of diagnosis. Nine (82%) of 11 lung transplant recipients who presented with capillaritis within 4 weeks post-transplant were alive at 1 year; all but 1 patient achieved expected percent predicted FEV1 values. Only 3 (14%) of 21 who presented with capillaritis > 1 month after transplant had a >20% decrease in the FEV1 after 12 months. These results suggest that post-transplant pulmonary capillaritis is (1) likely a form of acute allograft rejection clinically and histologically distinct from typical acute rejection, (2) less responsive to corticosteroid therapy than typical acute rejection, and (3) not associated with long-term adverse effects on allograft function. PMID- 16364856 TI - Donor total lung capacity predicts recipient total lung capacity after size reduced lung transplantation. AB - BACKGROUND: Size-reduced lung transplantation has only recently undergone widespread use, especially in highly urgent cases. However, it is still not considered standard procedure at most centers. It has the potential to alleviate the donor organ shortage by allowing the use of oversized grafts for small and pediatric recipients. Limited data exist on pre-operative parameters predicting functional outcome after lung transplantation in general, especially after size reduced lung transplantation. METHODS: All 98 patients undergoing primary lung transplantation during a 2-year period, including 27 size-reduced lung transplantations, were analyzed retrospectively. Pre-operative functional parameters were-after correction of estimated values according to the amount of size reduction-correlated with post-operative functional assessment. Actual and predicted total lung capacity (TLC) of transplant recipients and predicted TLC of donors was compared with the best post-operative TLC achieved within 12 months after transplantation. RESULTS: Size-reduced lung transplantation was performed in 27 cases. Downsizing was achieved by lobar transplantation (n = 9), split-lung transplantation (n = 2) or peripheral segmental resection (n = 16). There was a statistically highly significant (p < 0.01) correlation between donor TLC and best recipient TLC achieved after transplantation (Pearson's correlation coefficient = 0.675). No statistically significant correlation was seen between pre-operative recipient actual TLC and best post-operative TLC (p = 0.87; Pearson's correlation coefficient = 0.415). In standard lung transplant recipients post-operative TLC was correlated with both donor predicted TLC (p < 0.01; Pearson's correlation coefficient = 0.509) and actual pre-operative recipient TLC (p < 0.01; Pearson's correlation coefficient = 0.667). CONCLUSIONS: Post-operative recipient TLC in size-reduced lung transplantation can be predicted by donor TLC rather than pre-operative recipient TLC. PMID- 16364857 TI - Monitoring of Epstein-Barr viral load in pediatric heart and lung transplant recipients by real-time polymerase chain reaction. AB - BACKGROUND: Elevation in Epstein-Barr virus (EBV) load measured in peripheral blood has been proposed as a marker for development of post-transplant lymphoproliferative disease (PTLD), but there are few published data examining this relationship. We report the longitudinal surveillance of EBV for all recipients of heart (HTx), heart-lung (HLTx) and lung (LTx) transplants at our institution. METHODS: The study population included all patients transplanted between January 2003 and July 2004. EBV load was serially measured in peripheral blood by real-time polymerase chain reaction (PCR). Results were correlated with recipient pre-transplant EBV status and development of PTLD. RESULTS: Forty-four transplant operations were performed, including 33 HTx, 6 HLTx and 5 LTx. Thirty two (73%) of the patients were EBV seropositive pre-transplant. Nineteen (44%) pediatric recipients developed EB viremia, including 17 HTx, 1 HLTx and 1 LTx. Eleven (58%) of these patients were EBV seropositive pre-transplant. EBV was first detected at a median of 30.5 days (range 2 to 81) post-transplant. The median peak EBV load in that group was 10,099 copies/ml (range 5,935 to 255,466) whole blood. One patient with cystic fibrosis post-LTx developed PTLD localized in the colon. This patient was EBV seronegative pre-transplant; peak EBV load was 14,513 copies/ml. Acute infectious mononucleosis was seen in 1 case. Positive pre transplant EBV status did not predict post-transplant EB viremia (positive predictive value 0.03). CONCLUSIONS: Contrary to earlier reports, our data demonstrate that a high EBV load does not lead to PTLD early post-transplant. These results do not support the practice of pre-emptively reducing immunosuppression in patients with raised EBV load. PMID- 16364858 TI - Cytomegalovirus infection presenting as bronchial polyps in lung transplant recipients. AB - BACKGROUND: Cytomegalovirus (CMV) is the most important opportunistic pathogen in lung transplant recipients and is associated with direct and indirect morbidity. Infection or disease with CMV is often diagnosed through detection of the virus in the blood, culture of bronchoalveolar lavage fluid, or histologic examination of lung tissue obtained on transbronchial biopsy. Endobronchial lesions involving the virus have received little attention, however. METHODS: We reviewed the records of 76 endobronchial biopsies obtained at our institution from January 1999 through October 2004 and performed a detailed examination of the specimens and clinical histories for all patients with biopsy evidence of bronchial CMV infection. RESULTS: We identified three patients with endobronchial CMV infection, manifested in each case as an endobronchial polyp. On histologic examination of the endobronchial biopsy specimens, cells with cytopathic effects characteristic or suggestive of CMV infection were present in two cases, and immunohistochemical staining confirmed the presence of CMV in all three. All patients were clinically well with normal chest X-rays before biopsy. Two of three patients had multiple subsequent episodes of CMV disease. CONCLUSIONS: To our knowledge, this is the first report of endobronchial polypoid CMV disease in lung transplant recipients. Our results suggest that biopsy of endobronchial lesions in pulmonary allografts should be pursued to establish a definitive etiology. PMID- 16364859 TI - Intravenous ribavirin is a safe and cost-effective treatment for respiratory syncytial virus infection after lung transplantation. AB - BACKGROUND: Community-acquired viral infections, such as respiratory syncytial virus (RSV), represent a risk factor for bronchiolitis obliterans syndrome (BOS), the major limiting factor for long-term survival after lung transplantation (LTx). RSV often presents with acute bronchiolitis and may be fatal in 10% to 20% of patients. Standard therapies for RSV include nebulized ribavirin with or without steroids, but are costly and inconvenient. We investigated the utility of intravenous (IV) ribavirin with steroids for the treatment of RSV infection after LTx. METHODS: RSV was identified in nasopharyngeal and throat swabs (NPS) using indirect fluorescent antibody (IFA) testing in 18 symptomatic patients, which was confirmed by viral culture in 14. Data were collected for the period between April 2002 and October 2004. The study included 10 men and 8 women, mean age 42 +/- 15 (range 18 to 63) years. Transplant procedures were 5 single LTx and 13 bilateral LTx. RSV diagnosis was made on Day 1,374 +/- 1,270 (range 61 to 4,598, median 935) post-operatively. Underlying diagnoses included cystic fibrosis (n = 9), emphysema (n = 7) and pulmonary fibrosis (n = 2). All 18 patients received intravenous (IV) ribavirin (33 mg/kg on Day 1 and 20 mg/kg/day thereafter in 3 divided doses) with oral prednisolone (1 mg/kg) until repeat NPS were negative for RSV on IFA. Median therapy was 8 days (6 to 15). RESULTS: The mortality rate was 0%. Mean FEV1 fell from 2.1 +/- 1.0 liter (0.7 to 3.7 liters) to 1.8 +/- 0.9 liter (0.5 to 3.6 liters) (p < 0.001), but recovered to 2.1 +/- 0.9 (0.7 to 3.7 liters) within 3 months and was maintained at follow-up of 521 +/- 328 days (141 to 1,023 days, median 302). Only 1 patient developed bronchiolitis obliterans syndrome (BOS). Complications included mild hemolytic anemia (blood hemoglobin fell from 122 +/- 22 [84 to 154] g/liter to 107 +/- 18 [75 to 138] g/liter, p = 0.02). Cost savings per 8-day course were $US15,913 when compared with nebulized therapy at 6 g/day (p < 0.001). CONCLUSIONS: This is the largest reported series of treated RSV cases after LTx and the first to show that therapy with IV ribavirin and oral corticosteroids is well tolerated and effective. Cost utility vs nebulized therapy has been established. Early diagnosis and management are essential to prevent airway epithelial injury and subsequent BOS. PMID- 16364860 TI - Association between cyclosporine concentrations at 2 hours post-dose and clinical outcomes in de novo lung transplant recipients. AB - BACKGROUND: The objective of this study was to investigate the relationship between cyclosporine (CsA) pharmacokinetic parameters and clinical outcomes after lung transplantation. METHODS: Data from 48 lung or heart/lung transplant recipients originally recruited to a randomized, prospective clinical trial of Sandimmune vs Neoral and followed for 12 months were included in this study. CsA dosing was based on the trough concentration. CsA concentrations at 0 (C0), 2 (C2), and 6 (C6) hours post-dosing were obtained at 1, 2, 3, 4, 13, 26, 39, and 52 post-operative weeks. Based on their average C2 levels in the first post transplant month, patients were stratified retrospectively into Low C2 (<1,000 microg/liter, n = 18), Intermediate C2 (1,000-1,500 microg/liter, n = 16) and High C2 (>1,500 microg/liter, n = 14) Groups. RESULTS: Cyclosporine C2 was the best single-point determinant (r2 = 0.934) for area-under-the-concentration-time curve (AUC(0-6 hours)) compared with C0 (r2 = 0.267) or C6 (r2 = 0.304). The mean +/- SD values of CsA C2 and AUC(0 to 6 hours) in the first year post-transplant were significantly lower in patients with >2 rejection episodes compared with those with < or =2 rejection episodes (C2: 875 +/- 546 microg/liter vs 1,114 +/- 633 microg/liter, p = 0.01; AUC(0-6 hours): 4,036 +/- 1,904 microg x hour/liter vs 4,870 +/- 2,182 microg x hour/liter; p = 0.01) whereas C0 and C6 did not differ. Patients in the Intermediate C2 Group were free from rejection episodes for a significantly longer duration (p < 0.001) and had significantly higher predicted forced expiratory volume in 1 second (%) values (p < 0.001) compared with the Low and High C2 Groups. The percentage of increase in serum creatinine concentration by the end of first month post-transplant was significantly higher in the Intermediate C2 Group (p < 0.003). CONCLUSIONS: CsA C2 concentrations correlated better with the incidence of multiple rejections after lung transplantation than did C0 or C6. C2 concentrations between 1,000 and 1,500 microg/liter within the first post-operative month may be associated with better graft outcomes and improved pulmonary function and worsened renal function. PMID- 16364861 TI - Sirolimus in cardiac transplantation: use as a primary immunosuppressant in calcineurin inhibitor-induced nephrotoxicity. AB - BACKGROUND: Calcineurin inhibitor (CNI) immunosuppressants are a major cause of renal dysfunction in cardiac transplant recipients, leading to increased morbidity and mortality. The aim of this study was to evaluate the efficacy and safety of CNI withdrawal and substitution with sirolimus as the primary immunosuppressant, and assess the effect on renal function in cardiac transplant recipients with CNI-induced renal impairment. METHODS: Thirty-four stable cardiac transplant recipients (range 1 to 14 years post-transplant) with CNI-induced nephrotoxicity (iothalamate clearance 25 to 50 ml/min) or cardiac allograft vasculopathy (CAV) were enrolled. Twelve patients (Group A) were prospectively enrolled for renal dysfunction. The remaining patients (n = 22, Group B) were converted to sirolimus on clinical grounds because of poor renal function or the presence of CAV. CNI was withdrawn gradually over 12 weeks. Sirolimus was started at 1 mg/day with titration over 2 weeks to achieve levels of 10 to 15 ng/ml. Echocardiograms and cardiac biopsies were performed to determine rejection. Adjunct immunosuppression was left unchanged. Follow-up iothalamate clearance was performed. A further 24 patients (Group C) were retrospective controls, stable (range 2 to 10 years post-transplant), and maintained on a standard CNI-based immunosuppressant regimen. RESULTS: Iothalamate clearance (C(i)) improved significantly (Group A baseline: 36.08 +/- 2.4 ml/min to 48.67 +/- 4.1 ml/min, p = 0.004; Group B baseline: 48.14 +/- 3.2 ml/min to 55.77 +/- 4.2 ml/min, p < 0.001) without exacerbating rejection or compromising cardiac function. By contrast, in controls, Group C, the baseline renal clearance declined from 40.04 +/- 1.86 ml/min to 34.63 +/- 1.6 ml/min over the course of 1 year (p < 0.01). CONCLUSIONS: Substitution of CNIs with sirolimus in cardiac transplant recipients is safe and effective and leads to an improvement in renal function, without compromise in cardiac function and rejection. PMID- 16364862 TI - A randomized, controlled trial of C0- Vs C2-guided therapeutic drug monitoring of cyclosporine in stable heart transplant patients. AB - BACKGROUND: Cyclosporine monitoring using 2-hour post-dose samples (C2) is thought to be more efficacious than using pre-dose levels (C0) in managing immunosuppression for transplant patients. We evaluated the effect of C2 monitoring on cyclosporine dose and clinical parameters in stable heart transplant patients. METHODS: 125 stable heart transplant patients were randomized to C0 or C2 monitoring of cyclosporine levels for a period of six months. All patients had both C0 and C2 samples taken, and clinicians were blinded to one of the samples depending on randomization. The primary endpoint was the relative change in cyclosporine (Neoral) dose during the study period and secondary endpoints were change in creatine clearance, mortality, infection, and acute rejection. RESULTS: There was a significant decrease in the cyclosporine dose for the C2 group as compared with the C0 group (-11 mg/day and -26 mg/day respectively, p = 0.0025). No proven rejection episodes occurred in either group and there was no significant difference in the incidence of infection (C0 6, C2 10; p = 0.14), the change in renal function (change in creatine clearance C(0) +0.54 ml/min; C2 -0.16 ml/min; p = 0.61), the number of blood tests or dose adjustments between groups over the study period. Analysis of the blinded samples revealed that the reduction of cyclosporine dose in the C2 group could not be accounted for by reduced immunosuppression . CONCLUSION: C2 monitoring allows a significant cyclosporine dose reduction without compromising patient outcome in stable heart transplant patients. Further studies are required to ascertain whether this dose reduction can be translated into clinical benefit. PMID- 16364863 TI - The effect of beta-blocker use on cyclosporine level in cardiac transplant recipients. AB - BACKGROUND: Beta-blockers are frequently used after cardiac transplantation for blood pressure control. There is no well-known interaction between beta-blockers and cyclosporine A (CsA). However, recent reports have suggested that carvedilol, but not metoprolol, modulates P-glycoprotein (P-gp), a membrane protein that regulates CsA absorption. We evaluated the effects of carvedilol and metoprolol on CsA level when initiated in cardiac transplant recipients. METHODS: Using our cardiac transplant database, we identified patients who were started on either carvedilol or metoprolol for blood pressure control. We then compared their CsA doses and levels before and within 2 weeks after the initiation of beta-blocker therapy. RESULTS: We found 20 patients taking metoprolol and 12 patients taking carvedilol. With initiation of metoprolol, CsA level decreased in 12 patients and increased in 8 patients. The mean CsA level before and after metoprolol initiation was 236 ng/ml and 253 ng/ml, respectively (p = 0.50). In an attempt to maintain a therapeutic CsA level, the mean CsA dose was not significantly adjusted (from a mean of 293 mg/day to a mean of 294 mg/day; p = 0.92). In the Carvedilol Group, CsA level increased in 10 of 12 patients. The mean CsA level before the initiation of carvedilol was 257 ng/ml. The mean CsA level after carvedilol initiation was 380 ng/ml (p = 0.009). In an attempt to maintain a therapeutic CsA level, the mean CsA dose was reduced by 10%, from a mean of 319 mg/day to a mean of 288 mg/day (p = 0.004). CONCLUSION: Carvedilol, but not metoprolol, was associated with a significant increase in CsA levels after initiation in cardiac transplant recipients. Although carvedilol and CsA do not interact at the level of cytochrome P450 system, it appears that carvedilol influences CsA levels through its effects on P-gp. An average reduction of 10% is necessary on the CsA dose upon initiation of carvedilol, and close follow-up of the level is essential. PMID- 16364864 TI - Itraconazole prophylaxis in lung transplant recipients receiving tacrolimus (FK 506): efficacy and drug interaction. AB - BACKGROUND: Itraconazole is often given for fungal prophylaxis to lung transplant recipients after transplantation. The aim of this study was to determine the extent of interaction between tacrolimus and itraconazole in lung transplant recipients and the efficacy of itraconazole prophylaxis. METHODS: The study group included 40 lung transplant recipients followed for at least 12 months. All received prophylactic itraconazole, 200 mg twice a day, for the first 6 months after transplantation. Tacrolimus levels and dosage requirements were compared during and after itraconazole therapy. Rejection rate, fungal infection rate, and renal function were assessed. The mean cost per daily treatment of the itraconazole/tacrolimus combination and tacrolimus alone was calculated. RESULTS: The mean tacrolimus dose during itraconazole treatment was 3.26 +/- 2.1 mg/day compared with 5.74 +/- 2.9 mg/day after itraconazole was stopped (p < 0.0001) for a mean total daily dose elevation of tacrolimus of 76%. When the cost of itraconazole was taken into account, the average total daily cost of the combined treatment was US5.86 dollars less than the treatment with tacrolimus alone. No differences in the rejection or fungal infection rate, or in renal toxicity, were observed between the periods with and without itraconazole treatment, although less positive fungal isolates were identified during itraconazole therapy. CONCLUSION: Prophylaxis therapy with itraconazole is highly effective. Itraconazole reduces the dose of tacrolimus and therefore lowers the cost of therapy without causing an increase in rejection rate and with renal function preservation. PMID- 16364865 TI - Transforming growth factor-beta/interleukin-2-induced regulatory CD4+ T cells prolong cardiac allograft survival in rats. AB - BACKGROUND: Treatment of naive CD4+ T cells in vitro with transforming growth factor-beta (TGF-beta) or TGF-beta/interleukin-2 (IL-2), combined with stimulation in a mixed lymphoid culture (MLC), has been shown to generate CD4+ CD25+ regulatory T cells. However, little is known about the effect of these regulatory T cells on cardiac allograft survival in vivo. METHODS: CD4+ CD25+ T cells were generated from Lewis (LEW) rat spleen through a primary MLC with TGF beta (10 ng/ml) or TGF-beta/IL-2 (10 U/ml). The effect of adoptive transfer of the CD4+ CD25+ T cells (5.0 x 10(7)) was evaluated using an animal model of ACI rat cardiac allograft survival in LEW recipients. RESULTS: The MLC with TGF-beta or TGF-beta/IL-2 generated CD4+ CD25+ regulatory T cells, which suppressed the cytotoxic activity of LEW spleen T cells against irradiated ACI spleen cells in vitro. Adoptive transfer of the CD4+ CD25+ regulatory T cells intravenously to naive syngeneic recipients significantly prolonged the ACI cardiac allograft survival (N = 6, 13.5 +/- 3.4 days) compared with the control group (N = 6, 5.0 +/- 0.6 days). CONCLUSIONS: Intravenous administration of CD4+ CD25+ regulatory T cells, successfully generated by TGF-beta/IL-2 treatment, had a significant effect on cardiac allograft survival in this rat model. Adoptive transfer of regulatory T cells may represent a novel approach for preventing allograft rejection. PMID- 16364866 TI - Contribution of mesenchymal progenitor cells to tissue repair in rat cardiac allografts undergoing chronic rejection. AB - BACKGROUND: Mesenchymal progenitor cells (MPC) have recently been demonstrated to actively migrate into cardiac allografts during chronic rejection. This study examines the role of MPC in tissue repair of heart allografts in a rat model of chronic rejection. METHODS: The potential of a rat MPC line (Ap8c3) to differentiate to myofibroblasts and cardiomyocytes was studied in differentiation cultures. Ap8c3 cells tagged with an enhanced green fluorescent protein (eGFP) reporter gene were engrafted into Fischer 344 (F344) recipients of Lewis (LEW) cardiac allografts. Development of intragraft MPC into scar-forming fibroblasts and cardiomyocytes was studied using immunohistochemistry. RESULTS: Ap8c3 cells contain fibroblast progenitors (FP) positive for P07 antibody. Transforming growth factor (TGF)-beta stimulation promoted FP to terminally differentiate into myofibroblasts, which express alpha-smooth muscle actin (alphaSMA). In cardiac differentiation culture, Ap8c3 cells were induced by 5-azatiditin (5-aza) to form tropomyosin+ myotubes, and to express mRNA encoding for cardiac troponin I (TnI) and alpha-myosin heavy chain (alphaMHC). Transfusion of eGFP+ Ap8c3 cells to F344 recipients resulted in migration of eGFP(+) cells into LEW heart allografts, as well as homing of the eGFP+ MPC to bone marrow. The majority of eGFP+ cells in the heart allografts appeared to be vimentin-expressing fibroblasts. Foci of eGFP+ myocardium were also detected in all heart allografts, with eGFP+ cardiomyocytes representing 4.8 +/- 1.2% of the allografted eGFP+ cells. CONCLUSIONS: The data suggest that rat MPC participate in tissue repair in heart allografts by giving rise to scar-forming myofibroblasts and cardiomyocytes. PMID- 16364867 TI - Regulation of fibroblasts by activated and non-activated immune cells. AB - BACKGROUND: Rejection of transplanted tissue is characterized by cell infiltration and interstitial edema. Graft fibroblasts and fibroblast products are partly involved in the regulation of both these phenomena. Knowledge about the mechanisms behind fibroblast activation may lead to new strategies to prevent rejection. This study investigated whether cells of the immune system have the capacity to regulate fibroblast activation. METHODS: Fibroblasts isolated from rejecting heart transplants or from normal heart tissue were cultured in the presence of supernatants of stimulated or non-stimulated immune cells. The immune cells were challenged either in vitro (incubation with phytohemagglutinin) or in vivo (organ transplantation). Fibroblast proliferation and hyaluronan production were measured. RESULTS: Normal, sub-confluent heart fibroblasts showed an increased proliferation rate in the presence of supernatants of activated immunocompetent cells, irrespective of if these cells had been stimulated in vitro or in vivo. As expected, proliferation rate and hyaluronan production were upregulated in fibroblasts isolated from rejecting tissue. However, supernatants of biopsy specimens obtained from non-rejecting organs (syngeneic transplants or normal hearts) had an inhibitory effect on the growth rate of confluent fibroblasts isolated from rejecting tissue. CONCLUSIONS: We conclude that graft infiltrating cells and immune cells activated in vitro have the capacity to stimulate fibroblasts, most probably as a result of the production and secretion of fibroblast-stimulating factors. PMID- 16364868 TI - Identification of male cardiomyocytes of extracardiac origin in the hearts of women with male progeny: male fetal cell microchimerism of the heart. AB - BACKGROUND: Fetal progenitor cells may cross the placenta during pregnancy, persist for decades in the maternal bloodstream, and find a microenvironment conducive to colonization in a variety of maternal solid organs. Whether extracardiac fetal progenitors are present in the heart of women with male issue is unknown. METHODS: The hearts from 2 non-pregnant women who had given birth to 2 and 3 male children, respectively, were studied. Myocardial specimens from 2 men and 2 women (without history of pregnancies) were used as controls. Real time polymerase chain reaction was performed to amplify the SRY gene located at the Y chromosome. Fluorescence in situ hybridization (FISH) with probes specific for X and Y chromosomes was combined with alpha-actin immunohistochemistry to identify cardiac muscle cells. Histocompatibility studies were conducted in both patients and their male relatives. RESULTS: The SRY gene was amplified in the myocardium of both patients. FISH analysis showed clear evidence of male cells with the typical cardiomyocyte phenotype within the myocardium. X- and Y-chromosome bodies in the nuclei were found in 0.25% and 0.20% of cells, respectively. Increased human leukocyte antigen compatibility was observed between patients and their sons. CONCLUSIONS: This study identified male cardiomyocytes of extracardiac origin, presumably fetal, in the hearts of 2 women with male progeny. Fetal progenitor cells may colonize the heart and under appropriate microenvironmental stimuli, differentiate into cardiomyocytes. PMID- 16364869 TI - Phenotypic assessment of endothelial microparticles in patients with heart failure and after heart transplantation: switch from cell activation to apoptosis. AB - BACKGROUND: Endothelial microparticles (EMPs) are sub-microscopic membrane vesicles that are shed from the surface of endothelial cells during activation, injury and/or apoptosis. Endothelial cells release phenotypically and quantitatively distinct endothelial microparticles (EMPs) in activation and apoptosis. Therefore, the phenotypic assessment of EMPs can provide useful information reflecting the nature of endothelial injury. We tested the hypothesis that heart transplantation (HT) modifies the pattern of endothelial injury seen in patients with congestive heart failure (CHF). METHODS: Flow cytometry was used to measure EMPs identified by E-selectin (CD62) and platelet-endothelial cell adhesion molecule 1 (CD31) in 23 patients with advanced heart failure and in 23 HT recipients. A cohort of 23 healthy individuals served as controls. RESULTS: Heart failure patients were found to have significantly higher levels of EMP62E (577 counts/microl) than controls (192 counts/microl) and post-transplant patients (152 counts/microl) (p < 0.0001). Levels of endothelial microparticles expressing CD31 were significantly different among study groups (analysis of variance [ANOVA], p = 0.001). Heart failure patients had significantly higher levels (1,526 counts/microl) than controls (395 counts/microl) (p < 0.01). Levels of EMP31 remained elevated after heart transplant (935 counts/microl) (p = non significant). The EMP62/EMP31 ratio, an index of activation (high ratio) or apoptosis (low ratio), was significantly different between the groups (ANOVA, p = 0.01). Post-transplant patients had significantly lower ratios (0.16) than CHF patients (0.38) and controls (0.49). CONCLUSIONS: Cardiac transplantation is associated with a different pattern of endothelial cell injury than that seen in heart failure. The phenotypic assessment of EMPs in post-transplant patients is consistent with increased apoptotic activity. PMID- 16364870 TI - Comparison of soluble glycoprotein 130 and cardiac natriuretic peptides as long term predictors of heart failure progression. AB - BACKGROUND: In patients with heart failure, B-type natriuretic peptides (BNP, N BNP) and atrial natriuretic peptide (ANP) are established prognostic markers. However, circulating interleukin (IL)-6-related cytokines and soluble glycoprotein 130 (sgp130), their common subunit for signal transduction, are also increased. We hypothesized that levels of circulating sgp130 and cardiac peptides provide independent prediction of worsening pump failure in the long term. METHODS: A series of 76 patients (77% male, 54 ischemic and 17 nonischemic, left ventricular ejection fraction 22% +/- 7%) had blood samples drawn for assay of sgp130, oncostatin-M, N-ANP, N-BNP, and BNP. A composite end point of worsening pump failure (requiring hospitalization, intravenous therapy, or urgent heart transplantation) and pump failure death was used for follow-up. RESULTS: During follow-up (up to 7 years), rate of worsening pump failure was 22.3%, including death. N-ANP (5666 +/- 3100 vs 7850 +/- 12164 fmol/ml), N-BNP (278 +/- 284 vs 250 +/- 297 pmol/ml), and oncostatin-M (15 +/- 28 vs 16 +/- 63 pg/ml) were similar in those who incurred worsening pump failure and in others. Mean sgp130 levels were 389 +/- 123 ng/ml in patients who developed worsening heart failure (Group A) and 289 +/- 123 ng/ml in stable patients (Group B; p < 0.0001). Mean BNP was 567 +/- 774 pg/ml in Group A and 307 +/- 324 pg/ml in Group B (p < 0.05). By using a cutoff value of 286 ng/ml for gp130 in Kaplan-Meier analysis, we found that the rate of freedom from worsening heart failure was significantly higher in patients below compared with patients above this cutoff point (p = 0.03). In univariate and multivariate Cox regression analysis, only sgp130 emerged as statistically significant (p < 0.001). CONCLUSIONS: In addition to BNP, sgp130 could be useful in identifying patients at high risk for heart failure progression. PMID- 16364871 TI - Response of doxorubicin-induced cardiomyopathy to the current management strategy of heart failure. AB - BACKGROUND: Doxorubicin (D) (Adriamycin) is a potent and efficacious chemotherapeutic agent in the treatment of various forms of cancer, but its use has been limited by the development of cardiac toxicity. Historically, D-induced cardiomyopathy (CMP) has been refractory to therapy. We report our experience with this form of CMP at the University of Alabama at Birmingham. METHODS: Twenty five patients (20 women, 5 men) with a clinical diagnosis of D-CMP were referred to our program from 1990 to 2003. Patient data were extracted from office charts. RESULTS: Patients were followed-up for 71 +/- 58 months. On presentation, the average left ventricular ejection fraction (LVEF) was 26 +/- 9.2%, and 88% of patients were New York Heart Association (NYHA) Class III or IV. Patients were treated with angiotensin-converting enzyme inhibitors (ACEi; n = 23) or angiotensin-receptor blocker (ARB; n = 2), and 15 were treated with a combination of ACEi and beta-blockers (BB). With medical therapy, LVEF improved significantly (26 +/- 9.2% vs 35 +/- 16.5%, p = 0.022), as did the NYHA class (p < 0.003). All survivors (n = 19) were NYHA Class I or II with medical therapy, with 10 (53%) being Class I. In the group of patients treated with ACEi + BB, there was a statistically significant improvement in LVEF (26 +/- 10.0% vs 37 +/- 17.6%, p = 0.028), which not seen in the ACEi group, with a strong trend toward normalization of LV function (47% vs 10%, p = 0.054). CONCLUSIONS: In the current era of management of heart failure, D-CMP carries a better prognosis than previously described. Early addition of BB may further improve LVEF. PMID- 16364872 TI - Surgical ventricular remodeling for patients with clinically advanced congestive heart failure and severe left ventricular dysfunction. AB - BACKGROUND: Surgical ventricular remodeling (SVR) is an accepted therapy for post infarction ventricular remodeling. Current literature on SVR outcomes has focused on heterogeneous populations with regard to left ventricular function and New York Heart Association (NYHA) class. We assessed outcomes after SVR in patients with advanced congestive heart failure (CHF) (NYHA Class III/IV) and a pre operative ejection fraction (EF) < or =20%. METHODS: Data were analyzed for 51 consecutive SVR patients from January 2002 to June 2004. Cardiac catheterization, echocardiography and magnetic resonance imaging (MRI) identified 62.7% (32 of 51) of patients with an EF < or =20%, with the majority having an EF < or =15% (65.6%; 21 of 32). Cox regression analysis was performed to determine predictors of mortality in patients with an EF < or =20%. Follow-up was 100% (32 of 32) complete. RESULTS: Mean age was 61.9 +/- 10.3 (range 40 to 80) years with a male:female ratio of 27:5. Operative mortality was 6.3% (2 of 32). Twenty-two percent (7 of 32) had concomitant mitral valve procedures. Follow-up demonstrated a statistically significant improvement in left ventricular volumes and EF in survivors. Cox regression analysis identified the following to be significant predictors of mortality: pre-operative left ventricular end-systolic volume index >130 ml/m2; pre-operative diabetes; and intra-aortic balloon pump usage. Pre operatively, all patients (32 of 32) were categorized as NYHA Class III/IV, with 69% (22 of 32) improving to NYHA Class I/II at follow-up (p < 0.01). Survival did not differ statistically between patients with an EF < or =20% and an EF >20% (n = 19). CONCLUSIONS: Our results indicate that SVR improves left ventricular function and functional status for patients with advanced CHF and a pre-operative EF < or =20%. Therefore, SVR is a viable surgical alternative for patients with severe left ventricular dysfunction. PMID- 16364873 TI - Angiotensin II type 1 receptor antagonist protects ventricular and coronary endothelial function after 24-hour heart preservation. AB - BACKGROUND: Angiotensin II type 1 (AT1) receptor antagonists may enhance the cyclic guanosine monophosphate-nitric oxide system and thereby attenuate ventricular and coronary endothelial dysfunction after heart preservation. METHODS: We used an isolated rabbit heart preparation perfused with blood from a support rabbit. The rabbit heart was excised, stored for 24 hours, and then perfused with blood from a support rabbit that was treated with an AT1 receptor antagonist (telmisartan; 5 mg/kg) or solvent. We evaluated the cardiac output with the working preparation, and coronary blood flow and coronary endothelial function with the Langendorff preparation. In addition, we measured the serum nitric oxide level in the coronary effluent. RESULTS: The Telmisartan Group showed higher plasma angiotensin II levels (928.6 +/- 136.2 vs 271.6 +/- 81.6 pg/ml, p < 0.01), better cardiac output (116.2 +/- 5.4 vs 88.8 +/- 7.1 ml/min, p < 0.05), and higher coronary blood flow (25.0 +/- 2.2 vs 14.9 +/- 1.3 ml/min, p < 0.01). The coronary blood flow in response to acetylcholine was higher in the Telmisartan Group (47.8 +/- 3.9 vs 28.0 +/- 2.1 ml/min, p < 0.01), but there was no difference in response to sodium nitroprusside. The Telmisartan Group showed higher serum nitric oxide levels in the coronary effluent (33.9 +/- 4.6 vs 20.6 +/- 3.3 mumol/liter, p < 0.05). CONCLUSIONS: Treatment with the AT1 receptor antagonist improved ventricular and endothelial function after 24-hour heart preservation. These data imply that AT1 activation plays a critical role in reperfusion injury. AT1 receptor blockade may be a promising strategy for long term heart preservation. PMID- 16364874 TI - Trends in lung pH and PO2 after circulatory arrest: implications for non-heart beating donors and cell culture models of lung ischemia-reperfusion injury. AB - BACKGROUND: A better understanding of lung tissue environment after circulatory arrest would allow more accurate cell culture models to study ischemia reperfusion lung injury and facilitate retrieval of lungs from non-heart-beating donors. METHODS: To establish the time course of changes in pH and PO2 in lung tissue after circulatory arrest, 12 Sprague-Dawley rats were sacrificed. After sternotomy, pH and PO2 microelectrodes were inserted into the lungs and sealed by application of Focal Seal. Rats were maintained at normothermia (37 degrees C). Two groups of rats (n = 6 atelectatic, n = 6 room air-inflated) were followed for 4 hours after arrest, when lung tissue adenine nucleotide levels were measured by chromatography and cell death was quantified by trypan blue exclusion. Human umbilical vein endothelial cells underwent simulated ischemia and 6 hours of cold storage by replacement of culture medium with cold Perfadex. Interleukin (IL)-6 and IL-8 were measured in medium 21 hours later by enzyme-linked immunosorbent assay (ELISA). RESULTS: In both groups of rats, lung [H+] increased linearly with time. In atelectatic lungs, PO2 fell precipitously, but in inflated lungs, PO2 decreased linearly for 60 to 75 minutes post-mortem and then became stable. After 4 hours at 37 degrees C, most parenchymal lung cells were dead in both groups. IL 6 and IL-8 levels increased significantly in medium of cultured endothelial cells subjected to cold storage without hypoxia. CONCLUSIONS: In room-air-inflated lungs maintained at 37 degrees C, oxygen consumption continues for at least 1 hour after circulatory arrest. Warm atelectasis is poorly tolerated. Hypothermic storage can induce elaboration of cytokines by endothelial cells in the absence of hypoxia. PMID- 16364876 TI - The effect of anti-inflammatory properties of mycophenolate mofetil on the development of lung reperfusion injury. AB - BACKGROUND: Lung ischemia-reperfusion injury (LIRI) is associated with an increased incidence of both primary graft failure and obliterative bronchiolitis. The immunosuppressant mycophenolate mofetil (MMF) has recently been shown to attenuate inflammatory injury in acute ischemia-reperfusion models via a mechanism that is presently unclear. These experiments studied the effects of MMF in a warm, in situ LIRI model, focusing on transcriptional regulation of pro inflammatory mediators. METHODS: Left lungs of rats were rendered ischemic for 90 minutes and reperfused for up to 4 hours. Treated animals received 10 mg/kg of intravenous MMF at 2 hours before ischemia. Left lung injury was quantitated by myeloperoxidase (MPO) content, permeability indices and bronchoalveolar lavage (BAL) inflammatory cell counts. Lungs were analyzed by electrophoretic mobility shift assay (EMSA) for transcription factor transactivation and by enzyme-linked immunoassay for BAL chemokine protein content. RESULTS: MMF significantly reduced lung vascular permeability indices, MPO content and alveolar leukocyte counts at 4 hours of reperfusion. There was significant attenuation of activator protein 1 (AP-1) and early growth response 1 (EGR-1) transactivation, whereas nuclear factor-kappaB (NF-kappaB) was unaffected. Reductions in bronchoalveolar lavage monocyte chemoattractant protein 1 (MCP-1) and cytokine-induced neutrophil chemoattractant (CINC) protein content were found at 4 hours of reperfusion. CONCLUSIONS: MMF limits lung ischemia-reperfusion-induced increases in vascular permeability and inflammatory cell sequestration in lung parenchyma and alveolar spaces. The protection is mediated at the transcriptional level via an attenuation of early EGR-1 and AP-1 transactivation, which was found to be associated with reduced late MCP-1 and CINC protein secretion. The use of MMF in concert with an agent that affects NF-kappaB activation may provide even further protection against lung reperfusion injury as multiple inflammatory pathways are inhibited. PMID- 16364875 TI - S-nitroso human serum albumin attenuates ischemia/reperfusion injury after cardioplegic arrest in isolated rabbit hearts. AB - BACKGROUND: Depletion of nitric oxide (NO) is associated with ischemia/reperfusion injury. The novel NO donor, S-nitroso human serum albumin (S NO-HSA), could bridge NO depletion during reperfusion in cardiac transplantation and minimize ischemia/reperfusion injury. METHODS: In an isolated erythrocyte perfused working heart model, rabbit hearts were randomly assigned after assessment of hemodynamic baseline values to receive S-NO-HSA (0.2 micromol/100 ml, n = 8), L-arginine (10 mmol/100 ml, n = 8) or albumin (control) (0.2 micromol/100 ml, n = 8). After 20 minutes of infusion, the hearts were arrested and stored in Celsior (4 degrees C) enriched with respective drugs for 6 hours, followed by 75 minutes of reperfusion. Hemodynamic values were assessed and biopsy specimens were taken to determine calcium-ionophore stimulated release of NO and superoxide. RESULTS: During early reperfusion, recovery of cardiac output (75% +/- 6% vs 49% +/- 5%, p < 0.05) and coronary flow (99% +/- 8% vs 70% +/- 5%, p < 0.05) were higher, and myocardial oxygen consumption was reduced in the S-NO HSA Group compared with Control (4.08 +/- 0.46 ml/min/0.1 kg vs 6.78 +/- 0.38 ml/min/0.1 kg, p < 0.01). At the end of the experiment cardiac output (53% +/- 5% vs 27% +/- 5%, p < 0.01) was higher and left atrial pressure (115% +/- 9% vs 150% +/- 8%, p < 0.05) was lower in the S-NO-HSA Group compared with Control. NO release was increased (1,040 +/- 50 nmol/liter and 1,070 +/- 60 nmol/liter vs 860 +/- 10 nmol/liter, p < 0.01) and superoxide release diminished (31 +/- 5 nmol/liter and 38 +/- 5 nmol/liter vs 64 +/- 5 nmol/liter, p < .01) in the S-NO HSA and L-arginine Groups compared with Control. CONCLUSION: S-NO-HSA improved hemodynamic functions after prolonged hypothermic cardiac arrest by supplementing NO and thereby decreasing ischemia/reperfusion injury. PMID- 16364877 TI - Does Perfadex affect outcomes in clinical lung transplantation? AB - BACKGROUND: The use of a low-potassium-based preservation solution improves gas exchange in experimental models of lung transplantation. However, its efficacy in reducing the incidence of primary graft dysfunction (PGD) and improving patient outcomes in the clinical setting is controversial. METHODS: In this study we measured: oxygenation index (OI); International Society of Heart and Lung Transplantation (ISHLT) PGD grades; extubation times; intensive care unit (ICU) and hospital length of stay; 30-day, 90-day and 1-year survival rates; and bronchiolitis obliterans syndrome (BOS)-free survival. We compared 115 consecutive (2001 to 2004) lung recipients who received allografts preserved with Perfadex, a low-potassium dextran (LPD) solution, and compared the results with the previous 116 consecutive (1999 to 2001) lung recipients who received allografts preserved with modified Euro-Collins (MEC) solution. Recipients were classified as having severe PGD (ISHLT Grade III) if the lowest arterial oxygenation (P) to fraction of inspired oxygen (F) (P/F ratio) within 48 hours post-transplantation was <200. RESULTS: Baseline characteristics of the 2 cohorts were similar except for recipient age (LPD 53.5 vs MEC 49.9 years; p = 0.03). There were no differences in donor age, gender, category of transplant, indication for transplant, use of cardiopulmonary bypass or pre-operative pulmonary artery pressures. When gas-exchange parameters were measured upon arrival to the ICU (T0), at 24 hours post-transplant (T24) and at 48 hours post transplant (T48), the only significant finding was that the incidence of ISHLT Grade III PGD at T24 was lower in the LPD group compared with the MEC group (8% vs 20%, p = 0.03). The incidence of severe PGD at other timepoints was not statistically different (LPD vs MEC: T0, 17% vs 26%; T0 to T48, 25% vs 31%). Both groups had similar extubation rates at 48 hours post-transplant (LPD 64% vs MEC 67%). The 30-day survival (LPD 93% vs MEC 95%), 90-day survival (LPD 89% vs MEC 89%), 1-year patient survival (LPD 80% vs MEC 77%) and 1-year BOS-free survival (LPD 70% vs MEC 74%) were not statistically different. CONCLUSIONS: Lung preservation with LPD as compared with MEC does not improve early gas exchange or impact 90-day and 1-year mortality. Continued investigation into lung preservation solution composition is necessary to reduce the incidence of PGD. PMID- 16364878 TI - Glucocorticoid administration reduces cardiac dysfunction after brain death in pigs. AB - BACKGROUND: Traumatic brain injury and subsequent brain death (BD) account for nearly half of all organ donors, yet only 33% of available hearts are transplanted. Alterations in multiple physiologic pathways after BD can lead to cardiac dysfunction and exclusion from transplantation. Triple hormone resuscitation with methylprednisolone, thyroid hormone and vasopressin has had inconsistent results in the effort to reduce cardiac dysfunction associated with BD, but individual analysis of these agents is limited. The hypothesis was that glucocorticoid administration alone could reduce BD-associated cardiac dysfunction. METHODS: Crossbred pigs (25 to 35 kg) had BD induced by sub-dural balloon inflation. Hemodynamics were measured for 360 minutes after BD. Negative cerebral perfusion pressures and decreased laser Doppler cerebral blood flow confirmed BD. Animals (n = 5/treatment group) received: saline (Group 1); 30 mg/kg methylprednisolone 2 hours before BD (Group 2); or 30 mg/kg methylprednisolone 1 hour after BD (Group 3). Repeated measures analysis of variance and unpaired t-tests were used for appropriate comparisons. RESULTS: Left ventricular (LV) pre-load recruitable stroke work (PRSW) decreased in untreated Group 1 over time (p < 0.001), whereas PRSW in animals treated with glucocorticoids, Groups 2 and 3, was not different from baseline at 360 minutes after BD. Diastolic function measured as LV -dP/dt (minimum derivative of the change in pressure over time) and tau (time constant of isovolumic relaxation) was also preserved 360 minutes after brain death by glucocorticoids in Groups 2 and 3 (p > 0.05). Oxygen delivery 360 minutes after BD was higher in Group 2 compared with Group 1 (p = 0.02) and Group 3 (p = 0.006). CONCLUSIONS: Glucocorticoid therapy before or after BD preserved LV systolic and diastolic function. Glucocorticoids administered after brain death might increase the number of hearts available for transplant by reducing brain death-associated cardiac dysfunction. PMID- 16364879 TI - Who returns to work after heart transplantation? AB - BACKGROUND: Detailed information regarding the work history of heart transplant patients is limited. Therefore, the work history and factors associated with return to work at 1 year after heart transplantation were examined in 237 heart transplant patients as part of a longitudinal quality-of-life study at two university medical centers. Patient characteristics were as follows: 81% male; 89% white; mean age 54 years (range 24 to 71); mean level of education 13 years; and 84% were married. METHODS: Data were collected using the following instruments: Work History tool; Rating Question Form; Heart Transplant Stressor Scale; Quality of Life Index; Sickness Impact Profile; Jalowiec Coping Scale; Social Support Index; Heart Transplant Symptom Checklist; and Chart Review Form. Frequency distributions, chi-square, t-tests and stepwise regression were used to examine the work history of patients. RESULTS: Pre-transplant, only 17% of patients were working as compared with 26% (61 of 237) working by 1 year after transplant (p = 0.003). Pre-transplant non-working patients (n = 197) were hospitalized more frequently, were more physically disabled, had more symptom distress, and rated their health as poorer. After heart transplant non-working patients (n = 176) had more rejection, infection and medical complications and more hospital days. Patients who were working either pre- or post-transplant were more likely to hold jobs that were less physically demanding. Factors significantly associated with return to work by 1 year after heart transplant were better functional ability, higher education, fewer endocrine problems, fewer acute rejection episodes and shorter heart transplant waiting time. CONCLUSIONS: Clinical and demographic variables influence return to work after heart transplantation. Knowledge of these variables provides the health-care team with information to assist patients in securing gainful employment. PMID- 16364880 TI - Health-related quality of life in two hundred-eighty lung transplant recipients. AB - BACKGROUND: Health-related quality of life (HRQoL) has increasingly been accepted as a supplementary outcome measure for patients after lung transplantation (LTx). METHODS: Using a retrospective cross-sectional study design, 280 LTx recipients (3 months to 14 years after LTx) were asked to assess their HRQoL using a generic (Quality of Life Profile for Chronic Diseases) questionnaire, which was previously validated for this specific population. In addition, the questionnaire was also performed by 155 healthy participants. RESULTS: All sub-scale findings ranged from 2.40 to 3.08 (0 to 4) for all patients after LTx. A significantly reduced HRQoL was reported by the sub-cohort of patients living 5 to 6 years with the allograft for all sub-scales (p < 0.006), except for Social Functioning, and was associated with the incidence of bronchiolitis obliterans syndrome (BOS; p < 0.05). Cystic fibrosis patients (p < 0.05), single-lung transplant recipients (p < 0.05) and patients of older age (p < 0.05) showed significantly decreased physical ability ratings. Patients who remained free of infection and late acute rejection episodes scored themselves significantly higher with regard to their Relaxation Capabilities (p < 0.05 for rejection; p < 0.05 for infection) and Social Functioning (p < 0.01 for rejection; p < 0.05 for infection) vs patients who experienced infection or rejection episodes. Comparisons with a normative cohort showed similar HRQoL scales for LTx patients and the healthy population, except with regard to Social Functioning (p < 0.01). CONCLUSIONS: Self-ratings for HRQoL were high for all dimensions for the entire sample, and remained relatively similar even for medium- and long-term survivors. HRQoL was dependent on incidence of infections, rejections and the onset of BOS. Despite differences in life expectancy of LTx patients compared with the healthy population, HRQoL self-ratings were within similar ranges. PMID- 16364881 TI - Depression and quality of life in patients living 10 to 18 years beyond heart transplantation. AB - BACKGROUND: The purpose of this study was to advance current understanding of factors that influence long-term quality-of-life (QoL) outcomes after heart transplantation, by addressing the influence of depression on perceived health status. METHODS: Data were collected from all recipients (n = 137) still alive at >10 years after transplantation. They completed the Short Form Health Survey (SF 36) inventory and the Beck Depression Inventory (BDI) questionnaire, while objective measures of health status were retrieved from medical records. All instruments used had acceptable reliability and validity. Data were analyzed using descriptive statistics, general linear regression models and survival analysis. RESULTS: We assessed 137 patients who received transplants between November 1985 and June 1994 in Pavia and have survived 10 to 18 years after transplantation (mean 13.64 years, SD 2.25). They rated their health as good and only the physical QoL (PCS) was impaired when compared with the general population. Thirty-two percent of patients experienced mood depressive symptoms in the long term after transplantation, indicating a low perceived QoL. Higher educational qualification (p = 0.049), being unemployed and receiving a disability pension (p = 0.001), high triglycerides levels (p = 0.020) and lack of physical activity (p < 0.001) were predictors of high BDI scores. CONCLUSIONS: Assessment of depression levels and better understanding of risk factors for psychiatric disorders in the long term after transplantation could be of benefit in predicting negative outcomes and allowing future developments in patient management. PMID- 16364883 TI - Heart transplantation for progressive cardiomyopathy as a manifestation of MELAS syndrome. AB - Mitochondrial diseases represent a heterogeneous group of disorders associated with a wide array of clinical manifestations. The presentation of patients with mitochondrial pathology largely depends upon the dysfunction of organ systems with large metabolic/energy requirements, including cardiac, neurologic, and musculoskeletal. In particular, mitochondrial myocardial disease can be progressive resulting in congestive heart failure and end-stage heart disease. This article reviews the role of heart transplantation for a particular variant of mitochondrial disorder, mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome, and discusses perioperative management issues related to transplantation for mitochondrial cardiomyopathies. PMID- 16364882 TI - Predictors of physical functional disability at 5 to 6 years after heart transplantation. AB - BACKGROUND: Few reports were found in the literature that examined predictors of physical functional disability after heart transplantation. Therefore, the purpose of this study was to (1) describe physical functional disability, (2) identify differences in physical functional disability by demographic and clinical variables, and (3) identify predictors of physical functional disability at 5 to 6 years after heart transplantation. METHODS: A nonrandom sample of 311 patients (approximately 60 years of age, 78% male, and 90% Caucasian) who were 5 to 6 years post-heart transplantation were investigated. Patients completed 8 reliable and valid quality-of-life instruments via self-report. Data analyses included descriptive statistics, chi-square, independent t-tests, correlations, and stepwise multiple regression. Level of significance was set at p = 0.05. RESULTS: The level of physical functional disability was low at 5 to 6 years after heart transplantation, yet 59% of patients reported having physical disability. Women experienced more functional disability than men, and patients with comorbidities (i.e., diabetes mellitus and orthopedic problems) experienced more functional disability than patients without these comorbidities. At 5 to 6 years after heart transplantation, 70% of variance in physical functional disability was explained by activities of daily living, symptoms, comorbidities, psychologic status, and resource utilization variables. CONCLUSIONS: At 5 to 6 years after heart transplantation, most patients experienced low levels of physical functional disability. Differences in physical functional disability were identified by both demographic characteristics and clinical variables. Predictors of physical functional disability included activities of daily living and symptoms, and clinical, psychologic, and resource utilization variables. Knowledge of factors related to physical disability long-term after heart transplantation provides direction for the development of strategies to assist patients to reduce their level of disability or function adequately despite their disability. PMID- 16364884 TI - Aorta-to-left-atrial shunting through a previously performed heterotopic heart transplant: an unusual complication of orthotopic re-transplantation. AB - Heterotopic heart transplantation is rarely performed today, except in cases of recipient-donor size mismatch and when addressing complications associated with recipient pulmonary artery hypertension. As in orthotopic cardiac transplantation, occurrence of allograft vasculopathy occasionally calls for re transplantation. Although orthotopic re-transplantation without removal of the heterotopic allograft has been described previously, we report the case of a patient who presented with an unusual complication. PMID- 16364885 TI - Cardiac resynchronization therapy: a potential option for congenitally corrected transposition of the great vessels. AB - The use of cardiac resynchronization therapy in patients with QRS prolongation (left-sided interventricular conduction delay) and symptomatic (New York Heart Association class III and IV) heart failure despite optimal medical therapy is well established. This case report describes the use of cardiac resynchronization therapy to treat symptomatic congestive heart failure in 2 patients with congenitally corrected transposition of the great vessels. PMID- 16364886 TI - Disseminated Aspergillus fumigatus infection with consecutive mitral valve endocarditis in a lung transplant recipient. AB - Aspergillus infection is a known complication of lung transplantation and remains associated with high mortality rates. The manifestation of the infection varies from simple colonization of the lung to disseminated complicated infections. Early Aspergillus infection has been rarely observed in a small number of lung transplant recipients; most cases occur during the late post-operative period. The pulmonary involvement has often been described as the first clinical localization of the disease. Although other various forms of Aspergillus infection are not uncommonly encountered after lung transplantation, Aspergillus mitral valve endocarditis is rare. We present a case of disseminated Aspergillus fumigatus infection with consecutive mitral valve endocarditis having developed 78 days after double-lung transplantation for cystic fibrosis. PMID- 16364887 TI - Fatal Scopulariopsis infection in a lung transplant recipient: a case report. AB - A case of a fatal Scopulariopsis infection is reported in a 63-year-old lung transplantation patient with a 7-week history of dyspnea that presented initially with pericardial and pleural fluid. Because of a respiratory arrest, the patient was intubated and received positive pressure ventilation. Later, obstructive shock developed due to a pericardial tamponade requiring pericardectomy and then urgent extracorporal membrane oxygenation. The working hypothesis was of disseminated cytomegalovirus infection because cultures from bronchoalveolar lavage and transbronchial biopsy specimens were positive. The pericardial biopsy specimen showed fungal hyphae. After the patient died from the infection, the fungus was identified as Scopulariopsis acremonium. This case report describes the first case, to our knowledge, of an insidious Scopulariopsis acremonium infection in an immunocompromised lung transplant patient, underscoring the importance of a direct, invasive approach and early treatment with anti-fungal therapy in immunocompromised patients. PMID- 16364888 TI - Thrombolytic therapy due to thrombus formation associated with left ventricular assist devices. PMID- 16364889 TI - Hemodynamic response during non-pulsatile vs pulsatile left ventricular assist device support. PMID- 16364890 TI - To think or not to think: synaptic activity and Abeta release. AB - Accumulation of beta-amyloid protein (Abeta) in the extracellular space of the brain has been hypothesized to be a culprit in the pathogenesis of Alzheimer's disease. In this issue of Neuron, Cirrito et al. describe a series of experiments demonstrating that extracellular Abeta levels are directly modulated by neuronal and synaptic activity. PMID- 16364891 TI - Dystonia and the nuclear envelope. AB - Mutations in torsinA cause dominantly inherited early-onset torsion dystonia in humans. In this issue of Neuron, Goodchild et al. show that torsinA knockout and knockin mice have similar phenotypes, which suggests that the mutant torsinA allele causes disease because it has decreased function. The experiments also highlight the possible role of nuclear envelope dynamics in maintaining normal neuronal function. PMID- 16364892 TI - GABAergic neurons: gate masters of the amygdala, mastered by dopamine. AB - A hyperdopaminergic state, such as stress, is associated with an increase in affective behavior. In this issue of Neuron, Marowsky and colleagues identify two clusters of paracapsular intercalated GABA neurons in amygdala slice preparations of GAD67-GFP mice. These GABA neurons mediate inhibition from cortical afferents to both the major input and output station of the amygdala, are inhibited during action of dopamine via D1 receptors, and are thus likely to represent important cellular players during dopaminergic disinhibition related to increased affective behavior. PMID- 16364893 TI - Key regulators in neuronal polarity. AB - Neurons are highly polarized cells, most of which develop a single axon and several dendrites. These two compartments acquire specific characteristics that enable neurons to transmit intercellular signals from several dendrites to an axon. A wealth of recent studies has shown that PI 3-kinase, Rho family GTPases, the Par complex, and cytoskeleton-related proteins participate in the initial events of neuronal polarization. Here, we review the role of polarity-regulating molecules and the potential mechanisms underlying the specification of an axon and dendrites. PMID- 16364894 TI - Spinal muscular atrophy: a deficiency in a ubiquitous protein; a motor neuron specific disease. AB - Spinal muscular atrophy (SMA) is a neurodegenerative disease in humans and the most common genetic cause of infant mortality. The disease results in motor neuron loss and skeletal muscle atrophy. Despite a range of disease phenotypes, SMA is caused by mutations in a single gene, the Survival of Motor Neuron 1 (SMN1) gene. Recent advances have shed light on functions of the protein product of this gene and the pathophysiology of the disease, yet, fundamental questions remain. This review attempts to highlight some of the recent advances made in the understanding of the disease and how loss of the ubiquitously expressed survival of motor neurons (SMN) protein results in the SMA phenotype. Answers to some of the questions raised may ultimately result in a viable treatment for SMA. PMID- 16364895 TI - Mapping cortical activity elicited with electrical microstimulation using FMRI in the macaque. AB - Over the last two centuries, electrical microstimulation has been used to demonstrate causal links between neural activity and specific behaviors and cognitive functions. However, to establish these links it is imperative to characterize the cortical activity patterns that are elicited by stimulation locally around the electrode and in other functionally connected areas. We have developed a technique to record brain activity using the blood oxygen level dependent (BOLD) signal while applying electrical microstimulation to the primate brain. We find that the spread of activity around the electrode tip in macaque area V1 was larger than expected from calculations based on passive spread of current and therefore may reflect functional spread by way of horizontal connections. Consistent with this functional transynaptic spread we also obtained activation in expected projection sites in extrastriate visual areas, demonstrating the utility of our technique in uncovering in vivo functional connectivity maps. PMID- 16364896 TI - Synaptic activity regulates interstitial fluid amyloid-beta levels in vivo. AB - Aggregation of the amyloid-beta (Abeta) peptide in the extracellular space of the brain is central to Alzheimer's disease pathogenesis. Abeta aggregation is concentration dependent and brain region specific. Utilizing in vivo microdialysis concurrently with field potential recordings, we demonstrate that Abeta levels in the brain interstitial fluid are dynamically and directly influenced by synaptic activity on a timescale of minutes to hours. Using an acute brain slice model, we show that the rapid effects of synaptic activity on Abeta levels are primarily related to synaptic vesicle exocytosis. These results suggest that synaptic activity may modulate a neurodegenerative disease process, in this case by influencing Abeta metabolism and ultimately region-specific Abeta deposition. The findings also have important implications for treatment development. PMID- 16364897 TI - Loss of the dystonia-associated protein torsinA selectively disrupts the neuronal nuclear envelope. AB - An enigmatic feature of many genetic diseases is that mutations in widely expressed genes cause tissue-specific illness. One example is DYT1 dystonia, a neurodevelopmental disease caused by an in-frame deletion (Deltagag) in the gene encoding torsinA. Here we show that neurons from both torsinA null (Tor1a(-/-)) and homozygous disease mutant "knockin" mice (Tor1a(Deltagag/Deltagag)) contain severely abnormal nuclear membranes, although non-neuronal cell types appear normal. These membrane abnormalities develop in postmigratory embryonic neurons and subsequently worsen with further neuronal maturation, a finding evocative of the developmental dependence of DYT1 dystonia. These observations demonstrate that neurons have a unique requirement for nuclear envelope localized torsinA function and suggest that loss of this activity is a key molecular event in the pathogenesis of DYT1 dystonia. PMID- 16364898 TI - Hindbrain rhombic lip is comprised of discrete progenitor cell populations allocated by Pax6. AB - The lower rhombic lip (LRL) is a germinal zone in the dorsal hindbrain productive of tangentially migrating neurons, streaming extramurally (mossy fiber neurons) or intramurally (climbing fiber neurons). Here we show that LRL territory, operationally defined by Wnt1 expression, is parceled into molecular subdomains predictive of cell fate. Progressing dorsoventrally, Lmx1a and Gdf7 expression identifies the primordium for hindbrain choroid plexus epithelial cells; Math1, for mossy fiber neurons; and immediately ventral to Math1 yet within Wnt1(+) territory, a climbing fiber primordium dominated by Ngn1-expressing cells. Elimination of Pax6 results in expansion of this Ngn1(+) progenitor pool and reduction in the Math1(+) pool, with accompanying later enlargement of the climbing fiber nucleus and reductions in mossy fiber nuclei. Pax6 loss also disrupts Msx expression cell-nonautonomously, suggesting Pax6 may influence LRL progenitor identity indirectly through potentiating BMP signaling. These studies suggest that underlying the diversity and proportions of fates produced by the LRL is a precise suborganization regulated by Pax6. PMID- 16364899 TI - Distinct roles for secreted semaphorin signaling in spinal motor axon guidance. AB - Neuropilins, secreted semaphorin coreceptors, are expressed in discrete populations of spinal motor neurons, suggesting they provide critical guidance information for the establishment of functional motor circuitry. We show here that motor axon growth and guidance are impaired in the absence of Sema3A-Npn-1 signaling. Motor axons enter the limb precociously, showing that Sema3A controls the timing of motor axon in-growth to the limb. Lateral motor column (LMC) motor axons within spinal nerves are defasciculated as they grow toward the limb and converge in the plexus region. Medial and lateral LMC motor axons show dorso ventral guidance defects in the forelimb. In contrast, Sema3F-Npn-2 signaling guides the axons of a medial subset of LMC neurons to the ventral limb, but plays no major role in regulating their fasciculation. Thus, Sema3A-Npn-1 and Sema3F Npn-2 signaling control distinct steps of motor axon growth and guidance during the formation of spinal motor connections. PMID- 16364900 TI - The ion channel narrow abdomen is critical for neural output of the Drosophila circadian pacemaker. AB - Circadian clocks consist of transcriptional feedback loops housed in interdependent pacemaker neurons. Yet little is known about the neuronal output components essential for rhythmic behavior. Drosophila mutants of a putative ion channel, narrow abdomen (na), exhibit poor circadian rhythms and suppressed daylight activity. We find that NA is expressed in pacemaker neurons and induced expression within circadian neurons is sufficient to rescue these mutant phenotypes. Selective na rescue in distinct pacemaker neurons influences rhythmicity and timing of behavior. Oscillations of the clock protein PERIOD are intact in na mutants, indicating an output role. Pore residues are required for robust rescue consistent with NA action as an ion channel. In na mutants, expression of potassium currents and the key neuropeptide PDF are elevated, the latter consistent with reduced release. These data implicate NA and the pacemaker neural network in controlling phase and rhythmicity. PMID- 16364901 TI - Photoinactivation of native AMPA receptors reveals their real-time trafficking. AB - AMPA receptors mediate the majority of the fast excitatory transmission in the central nervous system. Much evidence suggests that the fast trafficking of AMPA receptors into and out of the postsynaptic membrane underlies changes in synaptic strength thought to be necessary for higher cognitive functions such as learning and memory. Despite the abundance of research conducted in this area, a direct, real-time functional assay that measures the trafficking of native AMPA receptors has been lacking. Toward this aim, we use a photoreactive, irreversible antagonist of AMPA receptors, ANQX, to rapidly silence surface AMPA receptors and investigate directly the trafficking of native AMPA receptors in real time. We find that the most dynamic movement of AMPA receptors occurs by lateral movement across the surface of neurons. Fast cycling of surface AMPA receptors with receptors from internal stores does occur but exclusively at extrasynaptic somatic sites. The cycling of synaptic AMPA receptors only occurs on a much longer timescale with complete exchange requiring at least 16 hr. This cycling is not dependent on protein synthesis or action potential driven network activity. These data suggest a revised model of AMPA receptor trafficking wherein a large internal store of AMPA receptors exchanges rapidly with extrasynaptic somatic AMPA receptors, and these newly inserted AMPA receptors then travel laterally along dendrites to reside stably at synapses. PMID- 16364902 TI - Physiologic diversity and development of intrinsically photosensitive retinal ganglion cells. AB - Intrinsically photosensitive retinal ganglion cells (ipRGCs) mediate numerous nonvisual phenomena, including entrainment of the circadian clock to light-dark cycles, pupillary light responsiveness, and light-regulated hormone release. We have applied multielectrode array recording to characterize murine ipRGCs. We find that all ipRGC photosensitivity is melanopsin dependent. At least three populations of ipRGCs are present in the postnatal day 8 (P8) murine retina: slow onset, sensitive, fast off (type I); slow onset, insensitive, slow off (type II); and rapid onset, sensitive, very slow off (type III). Recordings from adult rd/rd retinas reveal cells comparable to postnatal types II and III. Recordings from early postnatal retinas demonstrate intrinsic light responses from P0. Early light responses are transient and insensitive but by P6 show increased photosensitivity and persistence. These results demonstrate that ipRGCs are the first light-sensitive cells in the retina and suggest previously unappreciated diversity in this cell population. PMID- 16364903 TI - Photoreceptor adaptation in intrinsically photosensitive retinal ganglion cells. AB - A rare type of mammalian retinal ganglion cell (RGC) expresses the photopigment melanopsin and is a photoreceptor. These intrinsically photosensitive RGCs (ipRGCs) drive circadian-clock resetting, pupillary constriction, and other non image-forming photic responses. Both the light responses of ipRGCs and the behaviors they drive are remarkably sustained, raising the possibility that, unlike rods and cones, ipRGCs do not adjust their sensitivity according to lighting conditions ("adaptation"). We found, to the contrary, that ipRGC sensitivity is plastic, strongly influenced by lighting history. When exposed to a constant, bright background, the background-evoked response decayed, and responses to superimposed flashes grew in amplitude, indicating light adaptation. After extinction of a light-adapting background, sensitivity recovered progressively in darkness, indicating dark adaptation. Because these adjustments in sensitivity persisted when synapses were blocked, they constitute "photoreceptor adaptation" rather than "network adaptation." Implications for the mechanisms generating various non-image-forming visual responses are discussed. PMID- 16364904 TI - Adenosine and ATP link PCO2 to cortical excitability via pH. AB - In addition to affecting respiration and vascular tone, deviations from normal CO(2) alter pH, consciousness, and seizure propensity. Outside the brainstem, however, the mechanisms by which CO(2) levels modify neuronal function are unknown. In the hippocampal slice preparation, increasing CO(2), and thus decreasing pH, increased the extracellular concentration of the endogenous neuromodulator adenosine and inhibited excitatory synaptic transmission. These effects involve adenosine A(1) and ATP receptors and depend on decreased extracellular pH. In contrast, decreasing CO(2) levels reduced extracellular adenosine concentration and increased neuronal excitability via adenosine A(1) receptors, ATP receptors, and ecto-ATPase. Based on these studies, we propose that CO(2)-induced changes in neuronal function arise from a pH-dependent modulation of adenosine and ATP levels. These findings demonstrate a mechanism for the bidirectional effects of CO(2) on neuronal excitability in the forebrain. PMID- 16364905 TI - A specialized subclass of interneurons mediates dopaminergic facilitation of amygdala function. AB - The amygdala is under inhibitory control from the cortex through the activation of local GABAergic interneurons. This inhibition is greatly diminished during heightened emotional states due to dopamine release. However, dopamine excites most amygdala interneurons, suggesting that this dopaminergic gate may be mediated by an unknown subpopulation of interneurons. We hypothesized that this gate is mediated by paracapsular intercalated cells, a subset of interneurons that are innervated by both cortical and mesolimbic dopaminergic afferents. Using transgenic mice that express GFP in GABAergic interneurons, we show that paracapsular cells form a network surrounding the basolateral complex of the amygdala. We found that they provide feedforward inhibition into the basolateral and the central amygdala. Dopamine hyperpolarized paracapsular cells through D1 receptors and substantially suppressed their excitability, resulting in a disinhibition of the basolateral and central nuclei. Suppression of the paracapsular system by dopamine provides a compelling neural mechanism for the increased affective behavior observed during stress or other hyperdopaminergic states. PMID- 16364906 TI - Odorant representations are modulated by intra- but not interglomerular presynaptic inhibition of olfactory sensory neurons. AB - Input to the central nervous system from olfactory sensory neurons (OSNs) is modulated presynaptically. We investigated the functional organization of this inhibition and its role in odor coding by imaging neurotransmitter release from OSNs in slices and in vivo in mice expressing synaptopHluorin, an optical indicator of vesicle exocytosis. Release from OSNs was strongly suppressed by heterosynaptic, intraglomerular inhibition. In contrast, inhibitory connections between glomeruli mediated only weak lateral inhibition of OSN inputs in slices and did not do so in response to odorant stimulation in vivo. Blocking presynaptic inhibition in vivo increased the amplitude of odorant-evoked input to glomeruli but had little effect on spatial patterns of glomerular input. Thus, intraglomerular inhibition limits the strength of olfactory input to the CNS, whereas interglomerular inhibition plays little or no role. This organization allows for control of input sensitivity while maintaining the spatial maps of glomerular activity thought to encode odorant identity. PMID- 16364908 TI - Protein structure prediction: inroads to biology. AB - In recent years, there has been significant progress in the ability to predict the three-dimensional structure of proteins from their amino acid sequence. Progress has been due to new methods to extract the growing amount of information in sequence and structure databases and improved computational descriptions of protein energetics. This review summarizes recent advances in these areas and describes a number of novel biological applications made possible by structure prediction. Despite remaining challenges, protein structure prediction is becoming an extremely useful tool in understanding phenomena in modern molecular and cell biology. PMID- 16364907 TI - Integration of endocannabinoid and leptin signaling in an appetite-related neural circuit. AB - Recently developed therapeutics for obesity, targeted against cannabinoid receptors, result in decreased appetite and sustained weight loss. Prior studies have demonstrated CB1 receptors (CB1Rs) and leptin modulation of cannabinoid synthesis in hypothalamic neurons. Here, we show that depolarization of perifornical lateral hypothalamus (LH) neurons elicits a CB1R-mediated suppression of inhibition in local circuits thought to be involved in appetite and "natural reward." The depolarization-induced decrease in inhibitory tone to LH neurons is blocked by leptin. Leptin inhibits voltage-gated calcium channels in LH neurons via the activation of janus kinase 2 (JAK2) and of mitogen activated protein kinase (MAPK). Leptin-deficient mice are characterized by both an increase in steady-state voltage-gated calcium currents in LH neurons and a CB1R-mediated depolarization-induced suppression of inhibition that is 6-fold longer than that in littermate controls. Our data provide direct electrophysiological support for the involvement of endocannabinoids and leptin as modulators of hypothalamic circuits underlying motivational aspects of feeding behavior. PMID- 16364909 TI - The putative Drosophila transcription factor woc is required to prevent telomeric fusions. AB - Woc is a Drosophila zinc finger protein that shares homology with the human polypeptides ZNF261 and ZNF198 implicated in mental retardation and leukemia syndromes. We show that mutations in the woc gene cause frequent telomeric fusions in Drosophila brain cells. Woc localizes to all telomeres and most interbands of polytene chromosomes. In interbands, Woc precisely colocalizes with the initiating forms of RNA polymerase II (Pol II). To characterize the role of woc in telomere maintenance, we analyzed its relationships with Su(var)205, cav, atm, and rad50, four genes that prevent telomeric fusions; Su(var)205 and cav encode HP1 and HP1/ORC Associated Protein (HOAP), respectively. woc mutants displayed normal telomeric accumulations of both HP1 and HOAP, and mutations in cav, Su(var)205, atm, and rad50 did not affect Woc localization on polytene chromosome telomeres. Collectively, our results indicate that Woc is a transcription factor with a telomere-capping function independent of those of Su(var)205, cav, atm, and rad50. PMID- 16364910 TI - Oligomeric initiator protein-mediated DNA looping negatively regulates plasmid replication in vitro by preventing origin melting. AB - Although DNA looping between the initiator binding sites (iterons) of the replication origin (ori) of a plasmid and the iterons located in a cis-acting control sequence called inc has been postulated to promote negative control of plasmid DNA replication, not only was definitive evidence for such looping lacking, but also the detailed molecular mechanism of this control had not been elucidated. Here, we present direct evidence showing that both the monomeric and the dimeric forms of the RepE initiator protein of F factor together promote pairing of incC-oriF sites by DNA looping. By using a reconstituted replication system consisting of 26 purified proteins, we show further that the DNA loop formation negatively regulates plasmid replication by inhibiting the formation of an open complex at the replication origin, thus elucidating a key step of replication control. PMID- 16364911 TI - Structural analysis of the anaphase-promoting complex reveals multiple active sites and insights into polyubiquitylation. AB - The anaphase-promoting complex/cyclosome (APC/C) is an E3 ubiquitin ligase composed of approximately 13 distinct subunits required for progression through meiosis, mitosis, and the G1 phase of the cell cycle. Despite its central role in these processes, information concerning its composition and structure is limited. Here, we determined the structure of yeast APC/C by cryo-electron microscopy (cryo-EM). Docking of tetratricopeptide repeat (TPR)-containing subunits indicates that they likely form a scaffold-like outer shell, mediating assembly of the complex and providing potential binding sites for regulators and substrates. Quantitative determination of subunit stoichiometry indicates multiple copies of specific subunits, consistent with a total APC/C mass of approximately 1.7 MDa. Moreover, yeast APC/C forms both monomeric and dimeric species. Dimeric APC/C is a more active E3 ligase than the monomer, with greatly enhanced processivity. Our data suggest that multimerisation and/or the presence of multiple active sites facilitates the APC/C's ability to elongate polyubiquitin chains. PMID- 16364912 TI - Localization of the coactivator Cdh1 and the cullin subunit Apc2 in a cryo electron microscopy model of vertebrate APC/C. AB - The anaphase-promoting complex/cyclosome (APC/C) is a ubiquitin ligase with essential functions in mitosis, meiosis, and G1 phase of the cell cycle. APC/C recognizes substrates via coactivator proteins such as Cdh1, and bound substrates are ubiquitinated by E2 enzymes that interact with a hetero-dimer of the RING subunit Apc11 and the cullin Apc2. We have obtained three-dimensional (3D) models of human and Xenopus APC/C by angular reconstitution and random conical tilt (RCT) analyses of negatively stained cryo-electron microscopy (cryo-EM) preparations, have determined the masses of these particles by scanning transmission electron microscopy (STEM), and have mapped the locations of Cdh1 and Apc2. These proteins are located on the same side of the asymmetric APC/C, implying that this is where substrates are ubiquitinated. We have further identified a large flexible domain in APC/C that adopts a different orientation upon Cdh1 binding. Cdh1 may thus activate APC/C both by recruiting substrates and by inducing conformational changes. PMID- 16364913 TI - Cell motility is controlled by SF2/ASF through alternative splicing of the Ron protooncogene. AB - Ron, the tyrosine kinase receptor for the Macrophage-stimulating protein, is involved in cell dissociation, motility, and matrix invasion. DeltaRon, a constitutively active isoform that confers increased motility to expressing cells, is generated through the skipping of exon 11. We show that abnormal accumulation of DeltaRon mRNA occurs in breast and colon tumors. Skipping of exon 11 is controlled by a silencer and an enhancer of splicing located in the constitutive exon 12. The strength of the enhancer parallels the relative abundance of DeltaRon mRNA and depends on a sequence directly bound by splicing factor SF2/ASF. Overexpression and RNAi experiments demonstrate that SF2/ASF, by controlling the production of DeltaRon, activates epithelial to mesenchymal transition leading to cell locomotion. The effect of SF2/ASF overexpression is reverted by specific knockdown of DeltaRon mRNA. This demonstrates a direct link between SF2/ASF-regulated splicing and cell motility, an activity important for embryogenesis, tissue formation, and tumor metastasis. PMID- 16364914 TI - p38-dependent phosphorylation of the mRNA decay-promoting factor KSRP controls the stability of select myogenic transcripts. AB - Transcriptional and posttranscriptional processes regulate expression of genetic networks in response to environmental cues. The extracellular signal-activated p38 MAP kinase (p38) pathway plays a fundamental role in conversion of myoblasts to differentiated myocytes. p38 phosphorylates specific transcription factors and chromatin-associated proteins promoting assembly of the myogenic transcriptome. Here, we demonstrate that p38 alpha and beta isoforms also control muscle-gene expression posttranscriptionally, by stabilizing critical myogenic transcripts. KSRP, an important factor for AU-rich element (ARE)-directed mRNA decay, undergoes p38-dependent phosphorylation during muscle differentiation. KSRP phosphorylated by p38 displays compromised binding to ARE-containing transcripts and fails to promote their rapid decay, although it retains the ability to interact with the mRNA degradation machinery. Overexpression of KSRP selectively impairs induction of ARE-containing early myogenic transcripts, without affecting p38-mediated transcriptional responses. Our results uncover an unanticipated role for KSRP in establishing a biochemical link between differentiation-activated p38 signaling and turnover of myogenic mRNAs. PMID- 16364915 TI - Multiple processing body factors and the ARE binding protein TTP activate mRNA decapping. AB - Decapping is a key step in mRNA turnover. However, the composition and regulation of the human decapping complex is poorly understood. Here, we identify three proteins that exist in complex with the decapping enzyme subunits hDcp2 and hDcp1: hEdc3, Rck/p54, and a protein in decapping we name Hedls. Hedls is important in decapping because it enhances the activity of the catalytic hDcp2 subunit and promotes complex formation between hDcp2 and hDcp1. Specific decapping factors interact with the mRNA decay activators hUpf1 and TTP, and TTP enhances decapping of a target AU-rich element (ARE) RNA in vitro. Each decapping protein localizes in cytoplasmic processing bodies (PBs), and overexpression of Hedls produces aberrant PBs and concomitant accumulation of a deadenylated ARE mediated mRNA decay intermediate. These observations suggest that multiple proteins involved in human decapping are important subunits of PBs and are activated on ARE-mRNAs by the protein TTP. PMID- 16364916 TI - Molecular basis for bacterial class I release factor methylation by PrmC. AB - Class I release factors bind to ribosomes in response to stop codons and trigger peptidyl-tRNA hydrolysis at the P site. Prokaryotic and eukaryotic RFs share one motif: a GGQ tripeptide positioned in a loop at the end of a stem region that interacts with the ribosomal peptidyl transferase center. The glutamine side chain of this motif is specifically methylated in both prokaryotes and eukaryotes. Methylation in E. coli is due to PrmC and results in strong stimulation of peptide chain release. We have solved the crystal structure of the complex between E. coli RF1 and PrmC bound to the methyl donor product AdoHCy. Both the GGQ domain (domain 3) and the central region (domains 2 and 4) of RF1 interact with PrmC. Structural and mutagenic data indicate a compact conformation of RF1 that is unlike its conformation when it is bound to the ribosome but is similar to the crystal structure of the protein alone. PMID- 16364917 TI - The SAXS solution structure of RF1 differs from its crystal structure and is similar to its ribosome bound cryo-EM structure. AB - Bacterial class I release factors (RFs) are seen by cryo-electron microscopy (cryo-EM) to span the distance between the ribosomal decoding and peptidyl transferase centers during translation termination. The compact conformation of bacterial RF1 and RF2 observed in crystal structures will not span this distance, and large structural rearrangements of RFs have been suggested to play an important role in termination. We have collected small-angle X-ray scattering (SAXS) data from E. coli RF1 and from a functionally active truncated RF1 derivative. Theoretical scattering curves, calculated from crystal and cryo-EM structures, were compared with the experimental data, and extensive analyses of alternative conformations were made. Low-resolution models were constructed ab initio, and by rigid-body refinement using RF1 domains. The SAXS data were compatible with the open cryo-EM conformation of ribosome bound RFs and incompatible with the crystal conformation. These conclusions obviate the need for assuming large conformational changes in RFs during termination. PMID- 16364919 TI - The role of docking interactions in mediating signaling input, output, and discrimination in the yeast MAPK network. AB - Cells use a network of mitogen-activated protein kinases (MAPKs) to coordinate responses to diverse extracellular signals. Here, we examine the role of docking interactions in determining connectivity of the yeast MAPKs Fus3 and Kss1. These closely related kinases are activated by the common upstream MAPK kinase Ste7 yet generate distinct output responses, mating and filamentous growth, respectively. We find that docking interactions are necessary for communication with the kinases and that they can encode subtle differences in pathway-specific input and output. The cell cycle arrest mediator Far1, a mating-specific substrate, has a docking motif that selectively binds Fus3. In contrast, the shared partner Ste7 has a promiscuous motif that binds both Fus3 and Kss1. Structural analysis reveals that Fus3 interacts with specific and promiscuous peptides in conformationally distinct modes. Induced fit recognition may allow docking peptides to achieve discrimination by exploiting subtle differences in kinase flexibility. PMID- 16364918 TI - Crystal structure of MC159 reveals molecular mechanism of DISC assembly and FLIP inhibition. AB - The death-inducing signaling complex (DISC) comprising Fas, Fas-associated death domain (FADD), and caspase-8/10 is assembled via homotypic associations between death domains (DDs) of Fas and FADD and between death effector domains (DEDs) of FADD and caspase-8/10. Caspase-8/10 and FLICE/caspase-8 inhibitory proteins (FLIPs) that inhibit caspase activation at the DISC level contain tandem DEDs. Here, we report the crystal structure of a viral FLIP, MC159, at 1.2 Angstroms resolution. It reveals a noncanonical fold of DED1, a dumbbell-shaped structure with rigidly associated DEDs and a different mode of interaction in the DD superfamily. Whereas the conserved hydrophobic patch of DED1 interacts with DED2, the corresponding region of DED2 mediates caspase-8 recruitment and contributes to DISC assembly. In contrast, MC159 cooperatively assembles with Fas and FADD via an extensive surface that encompasses the conserved charge triad. This interaction apparently competes with FADD self-association and disrupts higher order oligomerization required for caspase activation in the DISC. PMID- 16364920 TI - Wild-type and mutant B-RAF activate C-RAF through distinct mechanisms involving heterodimerization. AB - The protein kinase B-RAF is mutated in approximately 7% of human cancers. Most mutations are activating, but, surprisingly, a small number have reduced kinase activity. However, the latter can still stimulate cellular signaling through the MEK-ERK pathway because they activate the related family member C-RAF. We examine the mechanism underlying C-RAF activation by B-RAF. We show that C-RAF is activated in the cytosol in a RAS-independent manner that requires activation segment phosphorylation and binding of 14-3-3 to C-RAF. We show that wild-type B RAF forms a complex with C-RAF in a RAS-dependent manner, whereas the mutants bind independently of RAS. Importantly, we show that wild-type B-RAF can also activate C-RAF. Our data suggest that B-RAF activates C-RAF through a mechanism involving 14-3-3 mediated heterooligomerization and C-RAF transphosphorylation. Thus, we have identified a B-RAF-C-RAF-MEK-ERK cascade that signals not only in cancer but also in normal cells. PMID- 16364921 TI - Eaf3 chromodomain interaction with methylated H3-K36 links histone deacetylation to Pol II elongation. AB - Eaf3, a component of the NuA4 histone acetylase and Rpd3 histone deacetylase complexes, is important for the global pattern of histone acetylation in Saccharomyces cerevisiae. Preferential deacetylation of coding regions requires the Eaf3 chromodomain and H3-K36 methylation by Set2. The Eaf3 chromodomain interacts with methylated H3-K36 peptides, suggesting that this interaction leads to preferential association and histone deacetylation of the 3' portions of coding regions by the Rpd3 complex. However, the Eaf3 chromodomain and H3-K36 methylation do not significantly affect acetylation at promoters, suggesting that Eaf3 has a distinct function, presumably in the NuA4 complex. Lastly, Eaf3 inhibits internal initiation within mRNA coding regions in a manner similar to FACT and Spt6. Our results link the pattern of preferential deacetylation at coding regions to the underlying patterns of H3-K36 methylation and phosphorylation of the RNA polymerase II C-terminal domain, and ultimately to the mechanism by which repressive chromatin structure is restored after transcriptional elongation. PMID- 16364922 TI - Effects of benzyl and phenethyl isothiocyanate on P450s 2A6 and 2A13: potential for chemoprevention in smokers. AB - Isothiocyanates have been shown to be potent inhibitors of carcinogenesis in animals exposed to a number of chemical carcinogens including the tobacco specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). In this study the effects of benzyl isothiocyanate (BITC) and phenethyl isothiocyanate (PEITC), two naturally occuring isothiocyanates, on P450 2A6 and 2A13 were investigated. P450s 2A6 and 2A13 are thought to be the primary human P450 enzymes responsible for the in vivo metabolism of nicotine and NNK, respectively. In vitro, BITC and PEITC efficiently inhibited P450 2A6- and 2A13 mediated coumarin 7-hydroxylation. The inhibition of P450 2A6 and 2A13 by BITC was non-competitive with KI's of 4.1 and 1.3 microM, respectively. PEITC was a more potent inhibitor of both enzymes than BITC, with a KI of 0.37 microM for P450 2A6 and 0.03 microM for P450 2A13. P450 2A6-mediated metabolism of nicotine and P450 2A13-mediated alpha-hydroxylation of NNK were also inhibited significantly by these two isothiocyanates. Both BITC and PEITC were able to inactivate P450 2A6 and 2A13 in an NADPH-dependent manner potentially through the formation of adducts to the apoprotein. The potent inhibition of P450 2A6- and 2A13-mediated metabolisms together with the ability of BITC and PEITC to inactivate the enzymes suggests the possibility that these isothiocyanates could be developed as chemopreventive agents to protect smokers who are unwilling or unable to quit smoking against lung cancer. PMID- 16364923 TI - DNA damage levels in prostate cancer cases and controls. AB - This study used the alkaline Comet assay to evaluate whether basal or H2O2 induced DNA damage is associated with prostate cancer (CaP) risk. Using lymphocyte samples from 158 CaP cases and 128 controls, collected in an ongoing case-control study, our results showed that basal DNA damage did not differ between cases and controls. However, the H2O2-induced DNA damage level was significantly higher in incident cases (mean +/- SD; 6.61 +/- 4.43, n = 102) than controls (5.30 +/- 3.60, n = 128) or prevalent cases (4.47 +/- 3.19; n = 56). Incident cases with a positive smoking history had significantly higher H2O2 induced DNA damage than never-smokers (7.57 +/- 4.82 versus 4.52 +/- 2.40; P < 0.001). Above-median H2O2-induced DNA damage was associated with a 1.61-fold increase in CaP risk [95% confidence interval (CI) = 0.92-2.81], after adjustment for age, race, benign prostatic hyperplasia (BPH), smoking history and family history (FH). Using the lowest quartile of H2O2-induced DNA damage as the referent group, the adjusted ORs for the 25th, 50th and 75th quartiles were 0.90 (95% CI = 0.39-2.05), 1.06 (95% CI = 0.48-2.35) and 2.05 (95% CI = 0.96-4.37), respectively (P = 0.046, test for linear trend). The association between CaP and DNA damage was modified by age, smoking history, family history and body mass index. Our results suggest that DNA damage may be associated with CaP risk. However, larger case-control and follow-up studies are warranted to further evaluate the potential application of the alkaline Comet assay in CaP risk assessment and prevention. PMID- 16364924 TI - Prospective study of 8-oxo-7,8-dihydro-2'-deoxyguanosine excretion and the risk of lung cancer. AB - Oxidative damage to DNA may be important in carcinogenesis and a possible risk factor for lung cancer. The urinary excretion of products of damaged nucleotides in cellular pools or in DNA may be important biomarkers of exposure to relevant carcinogens reflecting the rate of damage in steady state and may predict cancer risk. Oxidation of guanine in DNA or the nucleotide pool may give rise to 8-oxo 7,8-dihydro-2'-deoxyguanosine (8-oxodG) for urinary excretion. Oxoguanine glycosylase (OGG1) is the base excision enzyme repairing 8-oxodG in DNA by release of 8-oxoguanine. In a nested case-cohort design we examined associations between urinary excretion of 8-oxodG and risk of lung cancer as well as potential interaction with the OGG1 Ser326Cys polymorphism in a population-based cohort of 25 717 men and 27 972 women aged 50-64 years with 3-7 years follow-up. We included 260 cases with lung cancer and a sub-cohort of 263 individuals matched on sex, age and smoking duration for comparison. Urine collected at entry was analysed for 8-oxodG by HPLC with electrochemical detection. The excretion of 8 oxodG was higher in current smokers, whereas OGG1 genotype had no effect. Overall the incidence rate ratio (IRR) (95% confidence interval) of lung cancer was 0.99 (0.80-1.22) per doubling of 8-oxodG excretion and there was no interaction with OGG1 genotype. However, among never-smokers (eight cases and eight sub-cohort members) the IRR was 11.8 (1.21-115) per doubling of 8-oxodG excretion. The association between 8-oxodG excretion and lung cancer risk among never-smokers suggests that oxidative damage to DNA nucleotides is important in this group. PMID- 16364925 TI - Overexpression of thymosin beta-4 renders SW480 colon carcinoma cells more resistant to apoptosis triggered by FasL and two topoisomerase II inhibitors via downregulating Fas and upregulating Survivin expression, respectively. AB - The present work was conducted to further examine the effects of thymosin beta-4 (Tbeta4) upregulation on the apoptosis of SW480 colon cancer cells induced by T cells and various chemotherapeutic agents because reduced susceptibility to the cytotoxicity of an anti-Fas IgM (CH-11) in Tbeta4-overexpressing cells has previously been reported by us. As expected, Tbeta4 overexpressers were also more resistant to the killing effect of FasL-bearing Jurkat T cells. On the other hand, pretreating these cells with an MMP inhibitor restored not only their Fas levels but also their sensitivity to CH-11, suggesting a pivotal role of MMP in downregulating Fas in Tbeta4 overexpressers. Interestingly, while the susceptibilities of Tbeta4 overexpressers to 5-FU and irinotecan remained unchanged, they were more resistant to doxorubicin and etoposide which triggered apoptosis via a mitochondrial pathway. Concordantly, activation of both caspases 9 and 3 in Tbeta4 overexpressers by the two aforementioned topoisomerase II inhibitors was dramatically abrogated which could be accounted mainly by an increased expression of Survivin, a critical anti-apoptotic factor. Finally, poor survival was found in stage III colon cancer patients whose tumors were stained positively by the anti-Survivin antibody. Thus, advantages such as immune evasion and resistance to anticancer drug-induced apoptosis acquired by colon cancer cells through Tbeta4 overexpression might facilitate their survival during metastasis and chemotherapy. PMID- 16364927 TI - Clarification of retraction. PMID- 16364928 TI - Practical threshold for micronucleated reticulocyte induction observed for low doses of mitomycin C, Ara-C and colchicine. AB - Micronucleus induction was studied for the DNA target clastogens mitomycin C (MMC) and 1-beta-D-arabinofuranosylcytosine (Ara-C), and also the non-DNA target aneugen colchicine (COL) in order to evaluate the dose-response relationship at very low dose levels. The acridine orange (AO) supravital staining method was used for microscopy and the anti-CD71-FITC based method was used for flow cytometric analysis. In the AO method, 2000 reticulocytes were analysed as commonly advised, but in the flow cytometric method, 2000, 20,000, 200,000 and 1,000,000 reticulocytes were analysed for each sample to increase the detecting power (i.e. sensitivity) of the assay. The present data show that increasing the number of cells scored increases the statistical power of the assay when the cell was considered as a statistical unit. Even so, statistically significant differences from respective vehicle controls were not observed at the lowest dose level for MMC and Ara-C, or the lower four dose levels for COL, even after one million cells were analysed. When the animal was considered as a statistical unit, only the top dose group for each chemical showed significant increase of micronucleated reticulocytes frequency. As non-linear dose-response curves were obtained for each of the three chemicals studied, these observations provide evidence for the existence of a practical threshold for the DNA target clastogens as well as the non-DNA target aneugen studied. PMID- 16364929 TI - Non-invasive ventilation in elderly patients with acute exacerbations of COPD: bringing pressure to bear. PMID- 16364930 TI - Systematic review of definitions and methods of measuring falls in randomised controlled fall prevention trials. AB - OBJECTIVE: to review systematically the range of case definitions and methods used to measure falls in randomised controlled trials. DESIGN/METHODS: a Cochrane review of fall prevention interventions was used to identify fall definitions in published trials. Secondary searches of various databases were used to identify additional methodological or theoretical papers. Two independent reviewers undertook data extraction, with adjudication by a third reviewer in cases of disagreement. SETTINGS: community-dwelling and institutionalised older persons. RESULTS: 90 publications met the predefined inclusion criteria. Of these, 44 provided no definition of the term fall. In the remainder, there were substantial variations in the definition and methods of measuring falls. Reporting periods ranged from 1 week to 4 years with only 41% using prospective data collection methods. CONCLUSION: the standard of reporting falls in published trials is poor and significantly impedes comparison between studies. The review has been used to inform an international consensus exercise to make recommendations for a core set of outcome measures for fall prevention trials. PMID- 16364931 TI - Co-morbidity in older patients with COPD--its impact on health service utilisation and quality of life, a community study. AB - BACKGROUND: co-morbidity has been shown to be an important consideration in COPD with an estimated prevalence of 84%. In the Netherlands, a weak association between health-related quality of life and lung function has been found, with a closer link to co-morbidity. OBJECTIVE: to determine the influence of co morbidity on quality of life and health service utilisation in older patients with COPD in the community. DESIGN: observational cohort study. SETTING: general practice in the North East of England that has a list size of 8300. PARTICIPANTS: 27 patients aged 70 years or above on the practice COPD register. MEASUREMENTS: data on age and sex, spirometry to confirm the diagnosis of COPD, questionnaires to assess quality of life, activities of daily living (ADLs) and co-morbidity. Health service utilisation was recorded by the number of primary and secondary care attendances in the previous year. RESULTS: 10 had mild, 12 had moderate, and 5 had severe disease. Mean age was 76 years. Quality of life (QOL), co-morbidity and health service utilisation measurements were not significantly different between COPD severity groups. There was a significant positive correlation between increasing co-morbidity and poor QOL (r = 0.45, P < 0.05), and significant negative correlation between co-morbidity and ADL scores (scored inversely), r = -0.54, P < 0.05. Significant negative correlation was found between co-morbidity and primary care attendances (r = -0.41, P < 0.05) and significant positive correlation between worsening QOL and secondary care attendances (r = 0.46, P < 0.05). CONCLUSIONS: co-morbidity has an important part to play in COPD assessment, more accurately reflecting QOL in our population. Health service utilisation did not correlate to forced expiratory volume (FEV1) defined COPD severity. PMID- 16364932 TI - Low bone mineral density measurements in care home residents--a treatable cause of fractures. AB - PURPOSE: to assess predictors of fracture risk and treatment for osteoporosis among elderly care home residents. SUBJECTS AND METHODS: DESIGN: cross-sectional survey; SETTING: residents of care homes in Newcastle upon Tyne, UK; PARTICIPANTS: representative sample from residential care (87), nursing homes (105) and specialist homes for elderly people with dementia [elderly mentally infirm (EMI)]: residential (124) and nursing (76); MAIN OUTCOME MEASURES: dual energy X-ray absorptiometry bone mineral density (BMD) at calcaneum; functional assessments, including cognition, using Mini-Mental State Examination (MMSE), Clifton Assessment Procedure for the Elderly-Behaviour Rating Score (CAPE-BRS) and Functional Assessment Staging Test (FAST) scores; current drug prescription. RESULTS: MMSE, CAPE, FAST (all ANOVA P < 0.001) and weight (ANOVA P < 0.02) were lower in EMI homes. Drugs with sedative effects (chi-square, P < 0.0001) were more likely and calcium and vitamin D (CaD) supplementation (chi-square, P < 0.02) less likely in EMI care. For residential care, the odds ratio (OR) for sedative drugs in EMI was 2.13 (95% CI 1.11-4.06) with no significant difference between nursing homes. For CaD supplementation, the OR for EMI nursing homes was 0.19 (95% CI 0.05-0.72) and for EMI residential homes 0.38 (NS to 95% CI 0.12 1.27). BMD was low: mean T-score was -2.29 (95% CI -2-48 to -2.09) and Z-score 0.96 (95% CI -1.16 to -0.76) with a prevalence of osteoporosis (T-score < -1.6) of 69.2%. MMSE and FAST scores did not predict BMD. In EMI residential care, a decrease of CAPE score by 5 points was associated with a decrease in T-score by 0.6 (95% CI 0.15-1.1). CONCLUSIONS: of the tools used to assess function, only CAPE predicted low BMD in EMI residential care. Rates of CaD supplementation are particularly low in EMI care, where risk factors for fracture were the greatest. We conclude that fracture risk is neglected in these homes, and targeted education and treatment are warranted. PMID- 16364933 TI - The role of selective decontamination of the digestive tract in acute stroke. AB - BACKGROUND: following a stroke, morbidity and mortality is high, with aspiration pneumonia being a common complication. OBJECTIVES: to determine the levels of colonisation by and isolation of aerobic Gram-negative bacteria (AGNB) in acute stroke patients and determine the effect of selective decontamination of the digestive tract (SDD) on oral flora and whether it reduces both morbidity and mortality after an acute stroke. DESIGN: a prospective, randomised, placebo controlled double blind trial. SETTING: acute stroke assessment units of three hospitals in the northwest of England. SUBJECTS: 203 patients admitted to hospital following a first acute stroke. METHODS: participants were randomised to SDD oral gel or placebo. Swallow was assessed on admission to hospital, and oral swabs were obtained thrice weekly. Demographic and clinical data were recorded. RESULTS: 203 patients (106 males and 97 females) participated, of whom 20 died during their hospitalisation, 19 withdrew and full follow-up was obtained for the remaining 164. A total of 122 AGNB were isolated in 105 samples from 48 patients. Abnormal swallow on admission was found in 58 patients (29%). A total of 34 patients carried a single gram-negative micro-organism that was present on one or more occasions. More than one AGNB was carried in 14 patients, and organisms were significantly more likely to be isolated from the placebo group than the active group during weeks 2 and 3 of treatment (P = 0.034, chi-squared). Seven patients in the placebo group and one in the treatment group developed pneumonia (P = 0.029, Fisher's exact test). CONCLUSIONS: high carriage of and colonisation by AGNB was found within this study, which was reduced by the addition of SDD. Although SDD reduced the presence of both organisms and documented episodes of pneumonia, mortality remained unchanged. PMID- 16364934 TI - Tracheal pH monitoring and aspiration in acute stroke. AB - BACKGROUND: aspiration can lead to chest infections, increased morbidity and mortality in stroke sufferers. It is important clinically and for research purposes to identify all patients who aspirate. At present, videofluoroscopy is the gold standard for detecting aspiration. The aim of this study was to investigate aspiration in acute stroke patients, who are safe for oral intake as assessed by bedside swallow test and videofluoroscopy, using tracheal pH monitoring. METHODS: thirty-four stroke patients admitted to the Acute Stroke Unit gave informed consent and underwent tracheal pH monitoring 4-19 days post stroke. A standardised acid meal was served. RESULTS: two traces were discarded. Nine of the 32 remaining studies showed a drop in tracheal pH <5.5 following ingestion of an acidic meal. Two patterns of lowered tracheal pH were observed: three cases showed a prolonged fall in pH to <5.5, which took over 15 minutes to return to baseline and six had acute falls in pH to <5.5, which rapidly recovered in under 4 minutes. In six the drop occurred immediately after the meal, and in three a delay was observed prior to the drop. CONCLUSION: tracheal acidification, which could represent aspiration, has been observed in 9 of 32 stroke patients assessed as safe to take diet and fluids orally by bedside assessment and videofluoroscopy. This is a preliminary investigation that provides information about tracheal pH monitoring in acute stroke patients. PMID- 16364935 TI - Impaired cognitive performance in asymptomatic peripheral arterial disease: relation to C-reactive protein and D-dimer levels. AB - BACKGROUND AND PURPOSE: asymptomatic peripheral arterial disease (APAD), a highly prevalent condition in the general older population, is associated with an increased risk of cerebrovascular events because of co-existing clinical or subclinical cerebral atherosclerosis. The purpose of this study was to investigate whether cognitive function is impaired in stroke- and transient ischaemic attack-free patients with APAD, and whether inflammatory and haemostatic markers are associated independently with neuropsychological performance. METHODS: cognitive performances of 164 well-functioning, community dwelling patients with APAD were compared with those of 164 age-, gender- and education-matched healthy control subjects on six neuropsychological tests. Levels of C-reactive protein (CRP), D-dimer and fibrinogen were also analysed in all participants. RESULTS: patients with APAD scored significantly worse (P < 0.0001) than control subjects on five cognitive tests assessing domains of verbal working memory, attention, perceptuomotor speed, mental flexibility, visuoconstructive skills and visual memory. Multiple linear regression analyses showed that CRP and D-dimer were significant, independent predictors of poorer performances on four and three cognitive tests, respectively, within patients with APAD. CONCLUSIONS: patients with APAD show cognitive impairment in a range of psychometric tests, and CRP and D-dimer appear to be independent negative predictors of some cognitive performances. These findings suggest the need for screening for APAD among at-risk subjects in order to identify patients to be treated for prevention of functional decline and dementia. They also support the hypothesis that inflammation and hypercoagulability are implicated in the pathophysiology of cognitive dysfunction associated with APAD. PMID- 16364936 TI - Falls and health status in elderly women following second eye cataract surgery: a randomised controlled trial. AB - OBJECTIVE: first eye cataract surgery reduces the risk of falling. Most cataracts in elderly people are bilateral. Some binocular functions (e.g. stereopsis) are associated with falls. We aimed to determine if second eye cataract surgery reduces the risk of falling and to measure associated health gain. STUDY DESIGN AND SETTING: we randomised 239 women over 70, who had been referred to a hospital ophthalmology department, with one unoperated cataract, to expedited (approximately 4 weeks) or routine (12 months wait) surgery. Falls were ascertained by diary, with follow-up every 3 months for 12 months. We measured health status after 6 months. RESULTS: visual function (especially stereopsis) improved in the operated group. Over 12 months follow-up, 48 (40%) operated participants fell at least once, 22 (18%) fell more than once. Forty-one (34%) unoperated participants fell at least once, 22 (18%) fell more than once. Rate of falling was reduced by 32% in the operated group, but this was not statistically significant (rate ratio 0.68, 95% CI 0.39, 1.19, P = 0.18). Confidence, visual disability and handicap all improved in the operated compared with the control group. CONCLUSION: second eye cataract surgery improves visual disability and general health status. The effect on rate of falling remains uncertain. PMID- 16364937 TI - Factors associated with the initial acceptance of hip protectors amongst older people in residential care. PMID- 16364938 TI - Non-invasive ventilation for respiratory failure due to acute exacerbations of chronic obstructive pulmonary disease in older patients. PMID- 16364939 TI - The Checklist of Nonverbal Pain Indicators (CNPI): testing of reliability and validity in Norwegian nursing homes. PMID- 16364940 TI - Does admission grip strength predict length of stay in hospitalised older patients? PMID- 16364941 TI - Myasthenia gravis--a rare presentation with tongue atrophy and fasciculation. AB - We report the case of an unusual presentation of myasthenia gravis with tongue atrophy and fasciculation. Myasthenia gravis is an autoimmune condition associated with weakness and fatigability of voluntary muscles. In >50%, the initial symptoms and signs are related to extraocular muscle weakness, such as diplopia or ptosis [Tsung K, Seggev JS. An unusual cause of dysphagia. West J Med 1995; 163: 159-60]. Rarely, it is known to affect bulbar muscles and can lead to dysphagia and respiratory compromise. PMID- 16364942 TI - An unfamiliar course of a familiar illness: a rare ophthalmic complication of urinary sepsis. AB - Uncommon complications may occur in the context of illnesses encountered frequently in later life. They may be difficult to both identify and manage in a drowsy, confused patient, particularly in the absence of collateral information. We present an unusual ocular complication in a patient with systemic sepsis from a urinary tract infection, an unquestionably common diagnosis in older patients admitted to hospital. PMID- 16364943 TI - Inclusion body myositis: an underdiagnosed myopathy of older people. AB - Inclusion body myositis (IBM), a condition characterised by progressive muscle weakness and inclusion bodies visible on muscle biopsy, is the most common type of myopathy in patients over 50 years of age. However, it is not only under diagnosed but frequently misdiagnosed as polymyositis and hence wrongly treated with steroids. In the evaluation of progressive weakness in older Caucasian males, IBM should be an important diagnostic consideration. Treatment-resistant 'polymyositis' in patients over 50 years of age is often IBM. If there is no histological confirmation, the diagnostic criteria allow for a category of 'possible IBM'. Sometimes, the diagnosis is missed because of the slow progression of the disease and a lack of suspicion on the part of physicians. The following case report and literature review will explore many of these issues. PMID- 16364944 TI - An unusual cause of pleural effusion. AB - Primary effusion lymphoma (PEL) is a unique clinicopathological entity associated with human herpesvirus-8 (HHV-8) infection, occurring almost exclusively in human immunodeficiency virus (HIV)-infected individuals. We report a rare case of HHV-8 negative PEL in an HIV-negative elderly patient who presented with pleural effusion. The patient was treated with CHOP and Rituximab. As opposed to the general poor outcome of this disease, our patient achieved complete remission and is still without signs of disease 11 months after the last treatment. PMID- 16364945 TI - Unmeasured aspects of the geriatric day hospital! PMID- 16364946 TI - Tube feeding in patients with dementia. PMID- 16364947 TI - Re: Vitamin D for older people: how much, for whom and--above all--why? PMID- 16364948 TI - Temperature acclimation of photosynthesis: mechanisms involved in the changes in temperature dependence of photosynthetic rate. AB - Growth temperature alters temperature dependence of the photosynthetic rate (temperature acclimation). In many species, the optimal temperature that maximizes the photosynthetic rate increases with increasing growth temperature. In this minireview, mechanisms involved in changes in the photosynthesis temperature curve are discussed. Based on the biochemical model of photosynthesis, change in the photosynthesis-temperature curve is attributable to four factors: intercellular CO2 concentration, activation energy of the maximum rate of RuBP (ribulose-1,5-bisphosphate) carboxylation (Vc max), activation energy of the rate of RuBP regeneration (Jmax), and the ratio of Jmax to Vc max. In the survey, every species increased the activation energy of Vc max with increasing growth temperature. Other factors changed with growth temperature, but their responses were different among species. Among these factors, activation energy of Vc max may be the most important for the shift of optimal temperature of photosynthesis at ambient CO2 concentrations. Physiological and biochemical causes for the change in these parameters are discussed. PMID- 16364949 TI - Plant responses to potassium deficiencies: a role for potassium transport proteins. AB - The availability of potassium to the plant is highly variable, due to complex soil dynamics, which are strongly influenced by root-soil interactions. A low plant potassium status triggers expression of high affinity K+ transporters, up regulates some K+ channels, and activates signalling cascades, some of which are similar to those involved in wounding and other stress responses. The molecules that signal low K+ status in plants include reactive oxygen species and phytohormones, such as auxin, ethylene and jasmonic acid. Apart from up regulation of transport proteins and adjustment of metabolic processes, potassium deprivation triggers developmental responses in roots. All these acclimation strategies enable plants to survive and compete for nutrients in a dynamic environment with a variable availability of potassium. PMID- 16364950 TI - Is C4 photosynthesis less phenotypically plastic than C3 photosynthesis? AB - C4 photosynthesis is a complex specialization that enhances carbon gain in hot, often arid habitats where photorespiration rates can be high. Certain features unique to C4 photosynthesis may reduce the potential for phenotypic plasticity and photosynthetic acclimation to environmental change relative to what is possible with C3 photosynthesis. During acclimation, the structural and physiological integrity of the mesophyll-bundle sheath (M-BS) complex has to be maintained if C4 photosynthesis is to function efficiently in the new environment. Disruption of the M-BS structure could interfere with metabolic co ordination between the C3 and C4 cycles, decrease metabolite flow rate between the tissues, increase CO2 leakage from the bundle sheath, and slow enzyme activity. C4 plants have substantial acclimation potential, but in most cases lag behind the acclimation responses in C3 plants. For example, some C4 species are unable to maintain high quantum yields when grown in low-light conditions. Others fail to reduce carboxylase content in shade, leaving substantial over-capacity of Rubisco and PEP carboxylase in place. Shade-tolerant C4 grasses lack the capacity for maintaining a high state of photosynthetic induction following sunflecks, and thus may be poorly suited to exploit subsequent sunflecks compared with C3 species. In total, the evidence indicates that C4 photosynthesis is less phenotypically plastic than C3 photosynthesis, and this may contribute to the more restricted ecological and geographical distribution of C4 plants across the Earth. PMID- 16364952 TI - Advanced access: more than just GP waiting times? AB - BACKGROUND: Advanced access has been forwarded as a strategy for reducing waiting times in primary care; however, previous evaluations have raised important issues regarding its appropriateness. OBJECTIVES: The objectives of this paper are to assess the impact of advanced access on patient access to primary care services, and its broader effects on stakeholders. METHODS: A quantitative analysis of appointment data on 462 practices implementing advanced access, together with qualitative analysis of open survey responses and interviews with 28 practice staff. Appointment data recorded time to third available appointment for GP and practice nurse, together with the percentage of patients seen on their day of choice. Themes were identified from the interviews and survey responses and related to issues identified in previous research. RESULTS: The implementation of advanced access was associated with reductions in time to see practice nurses as well as GPs, and increases in the proportion of patients being seen on their day of choice. Interviewee and survey responses suggested that practice population characteristics may impact on the model, and some patient groups may be disadvantaged from the changes in the appointment systems seen in this study. Whilst experiences were mixed, the potential for broader changes to working practices of all practice staff was evident. CONCLUSIONS: In general, these results suggest that advanced access can have a positive impact across several aspects of primary care services, and not just the availability of GP appointments. However, it also highlights some problems, in that waiting times worsened in some practices and there were concerns that some vulnerable groups may be disadvantaged. PMID- 16364953 TI - Modulation of beta oscillations in the subthalamic area during motor imagery in Parkinson's disease. AB - Activation of the basal ganglia has been shown during the preparation and execution of movement. However, the extent to which the activation during movement is related to efferent processes or feedback-related motor control remains unclear. We used motor imagery (MI), which eliminates peripheral feedback, to further investigate the role of the subthalamic area in the feedforward organization of movement. We recorded local field potential (LPF) activity from the region of the subthalamic nucleus (STN) in eight patients with Parkinson's disease off dopaminergic medication during performance of a warned reaction time task. Patients were instructed to either extend the wrist [motor execution (ME)], to imagine performing the same task without any overt movement (MI), or, in a subgroup, to perform a non-motor visual imagery (VI) task. MI led to event-related desynchronization (ERD) of oscillatory beta activity in the region of the STN in all patients that was similar in frequency, time course and degree to the ERD occurring during ME. The degree of ERD during MI correlated with the ERD in trials of ME and, like ME, was accompanied by a decrease in cortico-STN coherence, so that STN LFP activity during MI was similar to that in ME. The ERD in ME and MI were both significantly larger than the ERD in VI. In contrast, event-related synchronization (ERS) was significantly smaller in trials of MI, and even smaller in trials of VI, than during ME. The data suggest that the activity in the region of the human STN indexed by the ERD during movement is related to the feedforward organization of movement and is relatively independent of peripheral feedback. In contrast, sensorimotor feedback is an important factor in the ERS occurring in the STN area after completion of movement, consistent with a role for this region in trial-to-trial motor learning or the re establishment of postural set following movements. PMID- 16364954 TI - Cortical spreading depression and peri-infarct depolarization in acutely injured human cerebral cortex. AB - Electrocorticographic (ECoG) activity was recorded for up to 129 h from 12 acutely brain-injured human patients using six platinum electrodes placed near foci of damaged cortical tissue. The method probes ECoG activity in the immediate vicinity of the injured cortex and in adjacent supposedly healthy tissue. Six out of twelve patients displayed a total of 73 spontaneous episodes of spreading depression of the ECoG. Of the remaining 6 patients 1 displayed an episode of synchronous depression of ECoG during surgery. Using the same electrodes we also measured the slow potential changes (SPC) (0.005-0.05 Hz) to test the hypothesis that the ECoG depressions were identical to Leao's cortical spreading depression (CSD), and to be able to record peri-infarct depolarisations (PIDs) in electrically 'silent' cortical tissue. Changes in the SPC indicate depolarization of brain tissue. For the analysis, the SPCs were enhanced by calculating the time integral of the ECoG signal. Spreading ECoG depressions were accompanied at every single recording site by stereotyped SPCs, which spread across the cortical mantle at 3.3 (0.41-10) mm/min (median, range), i.e. at the same speed of spread as the depression of the ECoG activity. The amplitude of the SPCs was 0.06-3 mV. In 4 out of 6 patients the ECoG recovered spontaneously. In 2 patients we subsequently recorded recurrent SPCs, but without recovery of the initial ECoG background activity until 2-5 h later. This represents the first direct recording of PIDs in acutely injured human brain. Evidence from this and our previous study of 14 brain-injured patients suggests that CSDs in acute brain disorders occur at higher incidence in patients <30 years (83%) than above (33%). CSD was recorded in 4 out of 5 traumatic brain injury patients, and in 2 out of 7 patients with spontaneous haemorrhages. We conclude that the spreading ECoG depressions recorded in patients are identical to CSDs recorded in animal experiments. We furthermore provide direct electrophysiological evidence for the existence of PIDs and hence a penumbra in the human brain. We hypothesize that the depolarization events might contribute to tissue damage in acute disorders in the human brain. PMID- 16364955 TI - Multimodal imaging of brain reorganization in motor areas of the contralesional hemisphere of well recovered patients after capsular stroke. AB - Clinical recovery after stroke can be significant and has been attributed to plastic reorganization and recruitment of novel areas previously not engaged in a given task. As equivocal results have been reported in studies using single imaging or electrophysiological methods, here we applied an integrative multimodal approach to a group of well-recovered chronic stroke patients (n = 11; aged 50-81 years) with left capsular lesions. Focal activation during recovered hand movements was assessed with EEG spectral analysis and H2(15)O-PET with EMG monitoring, cortico-cortical connectivity with EEG coherence analysis (cortico cortical coherence) and corticospinal connectivity with transcranial magnetic stimulation (TMS). As seen from comparisons with age-matched controls, our patients showed enhanced recruitment of the lateral premotor cortex of the lesioned hemisphere [Brodmann area (BA) 6], lateral premotor and to a lesser extent primary sensorimotor and parietal cortex of the contralesional hemisphere (CON-H; BA 4 and superior parietal lobule) and left cerebellum (patients versus controls, Z > 3.09). EEG coherence analysis showed that after stroke cortico cortical connections were reduced in the stroke hemisphere but relatively increased in the CON-H (ANOVA, contrast analysis, P < 0.05), suggesting a shift of functional connectivity towards the CON-H. Nevertheless, fast conducting corticospinal transmission originated exclusively from the lesioned hemisphere. No direct ipsilateral motor evoked potentials (MEPs) could be elicited with TMS over the contralesional primary motor cortex (iM1) in stroke patients. We conclude that (i) effective recovery is based on enhanced utilization of ipsi- and contralesional resources, (ii) basic corticospinal commands arise from the lesioned hemisphere without recruitment of ('latent') uncrossed corticospinal tract fibres and (iii) increased contralesional activity probably facilitates control of recovered motor function by operating at a higher-order processing level, similar to but not identical with the extended network concerned with complex movements in healthy subjects. PMID- 16364956 TI - Clinical, pathological and genetic characterization of hereditary sensory and autonomic neuropathy type 1 (HSAN I). AB - Hereditary sensory and autonomic neuropathy type I (HSAN I) is the most frequent type of hereditary neuropathy that primarily affects sensory neurons. The genetic locus for HSAN I has been mapped to chromosome 9q22.1-22.3 and recently the gene was identified as SPTLC1, encoding serine palmitoyltransferase, long chain base subunit-1. Sequencing in HSAN I families have previously identified mutations in exons 5, 6 and 13 of this gene. We analysed the SPTLC1 gene for mutations in 8 families with HSAN I, 60 individuals with sporadic sensory neuropathy, 6 HSAN II families, 20 Charcot-Marie-Tooth type I families and 20 families with Charcot Marie-Tooth type II. Six HSAN I families and a single sporadic neuropathy case had an identical SPTLC1 mutation. No mutations were found in the other groups. Genetic haplotyping across the HSAN I critical region in 5 families and the sporadic case suggested a common founder. Several characteristics, previously not widely recognized were identified, including lack of penetrance of the SPTLC1 mutation in some individuals, variability in age of onset along with an earlier age of onset in younger generations, in some patients surprisingly early and often severe motor involvement and an earlier onset characterized by motor involvement with demyelinating features in males compared to females in 4 families. The sensory findings were often disassociated with prominent pain and temperature loss. Neurophysiology mainly showed a sensory axonal neuropathy but in many individuals there was electrical evidence of demyelination. Sural nerve biopsies from six affected individuals and the post-mortem findings in 1 case showed mainly axonal loss. This in depth study on the phenotype of HSAN I in 6 families and a single sporadic case with a common founder identifies a number of poorly recognized features in this disorder and highlights the clinical heterogeneity both within and between families suggesting the influence of other genetic and acquired factors. PMID- 16364957 TI - Neuropsychological sequelae of bacterial and viral meningitis. AB - Survivors of meningitis often complain about neurological and neuropsychological consequences. In this study, the extent of these sequelae was quantified and correlated to MRI findings. Neurological, neuropsychological and neuroradiological examinations were performed with adult patients younger than 70 years, 1-12 years after recovery from bacterial meningitis (BM; n = 59), or from viral meningitis (VM; n = 59). Patients with other potential causes for neuropsychological deficits (e.g. alcoholism) were carefully excluded. Patients were compared to 30 healthy subjects adjusted for age, gender and length of school education. With the exception of attention functions, both patient groups showed more frequently pathological results than the control group for all domains examined. Applying an overall cognitive sum score, patients after BM did not differ significantly in their performance from patients after VM. Separate analyses of various cognitive domains, however, revealed a higher rate of persistent disturbances in short-term and working memory after BM than after VM. Moreover, patients after BM exhibited greater impairment of executive functions. Associative learning of verbal material was also reduced. These deficits could not be ascribed to impaired alertness functions or decreased motivation in BM patients. Applying a logistic regression model, the neuropsychological outcome was related to the neurological outcome. Patients with a Glasgow Outcome Scale (GOS) of <5 had more frequently impaired test results for non-verbal learning and memory. GOS was also correlated with performance in executive functions. Brain volume was lower and ventricular volume was higher in the bacterial than in the VM group, and cerebral volume and the amount of white matter lesions of patients after BM were negatively correlated with short-term and working memory. In conclusion, patients after both BM and VM with favourable outcome showed affected learning and memory functions. More patients after BM than after VM displayed pathological short-term and working memory. BM resulted in poorer performance in executive functions, language, short-term memory and verbal learning/memory tests. As a result of neurological and neuropsychological sequelae, BM with a GOS > or = 4 led to decreased activities of daily living but only a minority of patients were disabled in a way that social functions were affected. The extent of neuropsychological sequelae of BM might have been overestimated in earlier studies which often had not been controlled for comorbidity factors such as alcoholism. PMID- 16364958 TI - Myelin-laden macrophages are anti-inflammatory, consistent with foam cells in multiple sclerosis. AB - Multiple sclerosis lesion activity concurs with the extent of inflammation, demyelination and axonal suffering. Pro-inflammatory myeloid cells contribute to lesion development, but the self-limiting nature of lesions implies as yet unidentified anti-inflammatory mechanisms. We addressed the hypothesis that myelin ingestion by myeloid cells induces a foamy appearance and confers anti inflammatory function. First, we show that myelin-containing foam cells in multiple sclerosis lesions consistently express a series of anti-inflammatory molecules while lacking pro-inflammatory cytokines. Second, unique location dependent cytokine and membrane receptor expression profiles imply functional specialization allowing for differential responses to micro-environmental cues. A novel human in vitro model of foamy macrophages functionally confirmed that myelin ingestion induces an anti-inflammatory programme. Foamy macrophages are unable to respond to prototypical inflammatory stimuli but do express molecules involved in suppression of inflammation. These findings provide novel insights into the mechanisms of lesion control and may open new roads to intervention. PMID- 16364959 TI - Long-term clinical outcome of somatostatin analogues for treatment of progressive, metastatic, well-differentiated entero-pancreatic endocrine carcinoma. AB - BACKGROUND: Knowledge of factors able to predict the clinical outcome of homogenous series of entero-pancreatic endocrine tumours treated with somatostatin analogues is poor. This study was aimed at identifying predictors for efficacy of somatostatin analogues at inhibiting tumour growth and modifying patients' survival during long-term follow-up. PATIENTS AND METHODS: 31 patients with entero-pancreatic well-differentiated endocrine carcinoma received long acting somatostatin analogues. All had progressive, metastatic disease (87% liver metastases, 38.7% distant extra-hepatic metastases). RESULTS: Response rate after 6 months of treatment was 45.2% (all disease stabilisation: 27.8% of pancreatic vs. 81.8% of intestinal tumours, P = 0.007). The predictors for non-response were: pancreatic tumour (OR 5.8), no previous surgery (OR 6.7), and the presence of distant extra-hepatic metastases, the latter being also confirmed by multivariate analysis (OR 10.0). Responders maintained stabilisation for 26.5 months, and none died during follow-up. Different survival curves were observed for patients, responding at 6 months compared to non-responders (P = 0.004), 3 year survival rate being 100% and 52.3%, respectively. CONCLUSIONS: Distant extra hepatic metastases are the major predictor of poor efficacy of somatostatin analogues in progressive, metastatic, well-differentiated entero-pancreatic endocrine carcinomas. Patients achieving response after 6 months of treatment, maintain it throughout a long-term follow-up. Non-responders after 6 months of treatment, have a worse survival, and should be considered for alternative treatments. PMID- 16364960 TI - Novel formulations of taxanes: a review. Old wine in a new bottle? AB - Over the past two decades, the taxanes have played a significant role in the treatment of various malignancies. However, the poor solubility of these compounds necessitates the inclusion of surfactant vehicles in their commercial formulations. Cremophor EL and polysorbate 80 have long comprised the standard solvent system for paclitaxel and docetaxel, respectively. A number of pharmacologic and biologic effects related to both of these drug formulations have been described, including clinically relevant acute hypersensitivity reactions and peripheral neuropathy. In addition, these solvents affect the disposition of intravenously administered solubilized drugs and leach plasticizers from polyvinylchloride infusion sets. A number of strategies to develop formulations of surfactant-free taxanes have been developed. They include albumin nanoparticles, polyglutamates, taxane analogs and prodrugs, emulsions, and lipsomes. An overview of these novel formulations of taxanes, their mechanisms of action, pharmacokinetics, dose and administration, adverse effects, and clinical efficacy will be discussed. PMID- 16364961 TI - Hydroxyapatite-TiO2 hybrid coating on Ti implants. AB - A hydroxyapatite (HA)-titania (TiO(2)) hybrid coating is developed to improve the biocompatibility of titanium (Ti) implants. The HA predeposited layer on Ti via electron beam (e-beam) evaporation is subsequently treated by micro-arc oxidation (MAO) to produce an HA-TiO(2) hybrid layer on Ti. The e-beam-deposited HA layer has a thickness of approximately 1 microm and was highly dense prior to MAO. By means of MAO treatment, a rough and porous TiO(2) layer is formed beneath the HA layer with a simultaneous local dissolution of the HA layer. Due to the HA precoating, high concentrations of Ca and P are preserved on the coating surface. The osteoblast-like cells on the hybrid coating layer grow and spread favorably. The cell proliferation rate on the hybrid coatings is not much different from that on pure Ti or simple MAO-treated Ti. However, the alkaline phosphatase (ALP) activity of the cells is significantly higher (p < 0.05) on the HA-TiO(2) hybrid coatings than on either the pure Ti or the simple MAO-treated specimen, suggesting that the cellular activity on the hybrid coatings is improved. PMID- 16364962 TI - The effect of hyaluronic acid-supplemented bone graft in bone healing: experimental study in rabbits. AB - Hyaluronic acid (HA) is one of the essential components of extracellular matrix, which plays a predominant role in tissue morphogenesis, cell migration, differentiation, and adhesion. Bone allografts are frequently used to repair and reconstruct bone defects. In this study, two cavities of 3 mm diameter and depth have been created in the right tibia of 30 mature rabbits in accordance with the principles of general surgery. One of the cavities in the tibia is filled with HA and bone graft and the other is filled with only spongiosal bone graft, for the purpose of control. On the 20th, 30th, and 40th days, rabbits have been sacrificed in equal numbers and defective regions have been extracted. The Kruskal-Wallis test was applied to the data obtained in the result of histopathologic survey of specimens. In conclusion, the cavities that have been filled with HA and bone graft have shown higher scores than the control group during every period of the study. PMID- 16364963 TI - Synthesis and evaluation of novel bifunctional oligomer-based composites for dental applications. AB - Five novel bifunctional oligomers containing both carboxylic acid and methacrylate groups are synthesized, characterized, and used to formulate compomers by mixing with strontium fluoroaluminosilicate glass powder at a filler level of 75% (by weight). Compressive strength (CS) of the cements and viscosity of the resin liquids are used as screening tools to find the optimal formulation. Diametral tensile (DTS) and flexural strengths (FS) are also determined. Results show that the oligomers derivatized with glycerol dimethacrylate exhibit higher CS than those with 2-hydroxyethyl methacrylate. The CS increases with increasing diluent content, filler level, and light-exposure time. During aging, the cement shows an increase of strength over 24 h and then remains unaltered for up to 3 months. The experimental compomer is 45 and 69% higher in CS, 35 and 174% higher in DTS, and 39 and 170% higher in FS, respectively, as compared to Dyract and Fuji II LC. PMID- 16364964 TI - Chemical-physical characterization and in vitro preliminary biological assessment of hyaluronic acid benzyl ester-hydroxyapatite composite. AB - HYAFF11 is a biocompatible, biodegradable benzyl ester of hyaluronic acid. However, in order to use it for orthopedic application, its mechanical performance needs to be improved. In this study, a novel composite based on HYAFF11 polymer matrix reinforced with hydroxylapatite (HA) has been developed. Its advantage is having a similar component of the mineral phase of bone resulting in favorable osteoconductive properties. The present study has examined the compressive mechanical and surface chemical-physical properties of the novel HYAFF11-HA composite. Preliminary biological investigations, including pH and cytotoxicity studies of the material extracts, have also been performed using an in vitro primary human osteoblast-like cell model. Moreover, protein, especially fibronectin adsorption has been investigated following incubation in culture medium and human plasma. The results show a grainy surface topography composed mainly of C, P, and Ca, with a Ca/P atomic ratio indicating HA on the composite surface. Mechanical analysis shows an improvement of the compressive properties of HYAFF11 matrix, both in the dry and swollen states, with values in the range of that of spongy bone. No cytotoxic effects and no inhibition of cell proliferation have been observed in the presence of the material extracts with pH values within acceptable ranges for cell vitality. Protein studies reveal a similar pattern, but a higher amount of fibronectin following incubation in human plasma when compared with culture medium. The results show that the novel HYAFF11 HA composite shows a great potential for application in orthopedic fields, especially as vertebral trabecular bone substitute. PMID- 16364965 TI - In vivo performance of biodegradable calcium phosphate glass ceramics using the rabbit model: histological and SEM observation. AB - Two MK5 (45CaO-45P(2)O(5)-5MgO-5K(2)O, in mol%) and MT13 (45CaO-37P(2)O(5)-5MgO 13TiO(2), in mol%) glasses are prepared in the meta- and pyrophosphate regions and crystallized to obtain MK5B and MT13B, respectively. MK5B was obtained by controlled crystallization, and MT13B by powder sintering. As a result of these heat treatment processes, the crystalline phases precipitated in the glassy matrix are KCa(PO(3))(3), beta-Ca(PO(3))(2), beta-Ca(2)P(2)O(7) and Ca(4)P(6)O(19) phases for MK5B and CaTi(4)(PO(4))(6), TiP(2)O(7), alpha- and beta Ca(2)P(2)O(7) phases for MT13B. To assess the in vivo biological behavior of these glass ceramics, a mixed granulometry in the range 250-355 mum and 355-425 mum with a ratio of 1/1 was implanted for 2, 4, and 12 weeks in the tibiae of Japanese white rabbits. The results showed that the in vivo behavior was strongly affected by their solubility. All implanted materials, MK5B and MT13B, and beta tricalcium phosphate (beta-TCP) as control material, showed signs of degradation in vivo. However, the levels of degradation were quite different throughout the implantation periods. The highest degradation was observed for MK5B glass ceramic and the lowest for MT13B with beta-TCP in-between. All implanted materials allow for new bone formation in the bone defect area. At the longest implantation period (12 weeks), the MT13B and beta-TCP materials were almost completely surrounded by new bone tissue, whereas MK5B showed some unfilled spaces. This behavior is discussed in terms of the high degradation observed in previous studies. PMID- 16364966 TI - Modifications on collagen structures promoted by 1,4-dioxane improve thermal and biological properties of bovine pericardium as a biomaterial. AB - Collagen is a widely used raw material for biomaterial manufacture, which generally depends on chemical modifications of this fibrillar protein with cross linking agents to improve biocompatibility and mechanical properties. However, cross-linking reduces the natural properties of collagen, such as low immune response, low toxicity as well as the ability to promote cellular growth and attachment. In this work, the modifications promoted by 1,4-dioxane solvent on the collagen present in native bovine pericardium (NBP) matrix routinely used in bioprosthesis manufacture, with or without subsequent cross-linking by glutaraldehyde, has been studied. The structural changes of NBP evaluated by scanning electron microscopy show that 1,4-dioxane induces a more homogeneous material by increasing aggregation of collagen fibers, while transmission scanning electron microscopy shows that natural collagen fibril arrangement, integrity, and the D-periodicity pattern are maintained by solvent treatments. Measurements of thermal stability and resistance to collagenase enzymatic digestion of NBP matrices treated with 1,4-dioxane show an increase in melting temperature and decrease in biodegradability, as compared to native pericardium. Cross-linking with glutaraldehyde improves all the analyzed NBP properties, which are not impaired by previous treatment with 1,4-dioxane. Histological evaluation of NBP submitted to 1,4-dioxane treatment shows lower lipid and cell contents and improvement in other morphologic characteristics compared to native pericardium. Altogether, these results suggest the use of 1,4-dioxane organic solvent as an alternative non-cross-linking treatment for direct utilization on rich collagen matrices, resulting in materials with improved biocompatibility and physicochemical properties suitable for tissue engineering. PMID- 16364967 TI - Effect of ganciclovir on the hydrolytic degradation of poly(lactide-co-glycolide) microspheres. AB - Ganciclovir (GCV)-loaded poly(lactide-co-glycolide) (PLGA) microspheres, 125 +/- 11 mum in diameter, are produced using the emulsification/solvent evaporation technique. The release rate of the drug is studied for 20 weeks in a phosphate buffered solution of pH 7 at 37 degrees C. The release of the drug shows a triphasic release pattern, i.e., an initial burst, a diffusive phase, and a second burst. The initial burst occurs within the first 2 days of immersion. After the burst, the release is by diffusion for up to 13 weeks, followed by another burst release, which signals the onset of bulk degradation of the PLGA polymer. The presence of GCV molecules decreases the hydrolytic rate of PLGA degradation. Gel permeation chromatography (GPC), differential scanning calorimetry (DSC), field emission scanning electron microscopy (FESEM), and ultraviolet (UV) spectroscopy are used to assess the hydrolytic degradation and drug release rate of the microspheres. PMID- 16364968 TI - Accuracy of coalescent likelihood estimates: do we need more sites, more sequences, or more loci? AB - A computer simulation study has been made of the accuracy of estimates of Theta = 4Nemu from a sample from a single isolated population of finite size. The accuracies turn out to be well predicted by a formula developed by Fu and Li, who used optimistic assumptions. Their formulas are restated in terms of accuracy, defined here as the reciprocal of the squared coefficient of variation. This should be proportional to sample size when the entities sampled provide independent information. Using these formulas for accuracy, the sampling strategy for estimation of Theta can be investigated. Two models for cost have been used, a cost-per-base model and a cost-per-read model. The former would lead us to prefer to have a very large number of loci, each one base long. The latter, which is more realistic, causes us to prefer to have one read per locus and an optimum sample size which declines as costs of sampling organisms increase. For realistic values, the optimum sample size is 8 or fewer individuals. This is quite close to the results obtained by Pluzhnikov and Donnelly for a cost-per-base model, evaluating other estimators of Theta. It can be understood by considering that the resources spent collecting larger samples prevent us from considering more loci. An examination of the efficiency of Watterson's estimator of Theta was also made, and it was found to be reasonably efficient when the number of mutants per generation in the sequence in the whole population is less than 2.5. PMID- 16364969 TI - Evaluation of the Texas 0.08 BAC law. AB - AIMS: The purpose of the present study was to assess the effects on alcohol involved traffic crashes and fatalities of the 0.08 blood alcohol concentration (BAC) per se law introduced in the state of Texas in 1999. METHOD: Data pertaining to alcohol-involved traffic crashes and fatalities were extracted from two datasets: the Fatality Analysis Reporting System (FARS) compiled by the National Highway Traffic Safety Administration (for the period January 1995 September 2002), and the Texas Department of Public Safety (DPS) reports of Alcohol Related Motor Vehicle Traffic Accidents and Casualties (for the period January 1995-December 2000). The data were analysed using time-series methods (ARIMA routines). The effects of the law on all drivers were assessed, along with the effects among gender, racial, and age subgroups and crash location (urban vs rural). RESULTS: Separate time-series analyses were conducted with all alcohol involved and fatal alcohol-involved crashes from the DPS dataset and fatal alcohol-involved crashes from the FARS dataset as the outcome variables. None of the effects for either the total sample or any of the subgroups analysed was statistically significant (this was true of both the FARS and DPS datasets). CONCLUSIONS: While there is a growing body of evidence that indicates that 0.08 BAC laws can be effective in reducing alcohol-involved traffic accidents and fatalities, the present study shows that this was not the case in Texas. Future research should move beyond the simple question of whether or not 0.08 BAC laws 'work' and instead explore in more detail the conditions, such as publicity and enforcement, under which the law does or does not contribute to a decline in alcohol-involved accidents and fatalities. PMID- 16364970 TI - A late complication of a patent foramen ovale amplatzer occluder device. PMID- 16364971 TI - Incidence of renal dysfunction over 6 months in patients with chronic heart failure due to left ventricular systolic dysfunction: contributing factors and relationship to prognosis. AB - AIMS: To determine the prevalence and incidence of renal dysfunction (RD) in patients with chronic heart failure (CHF), to identify contributory factors and predictors of worsening renal function (WRF), and to explore the relationship between RD and mortality. METHODS AND RESULTS: Prospective data on 1216 patients with CHF were analysed. The glomerular filtration rate (GFR) was used to determine renal function, and WRF was defined as an increase in serum creatinine of >26.5 micromol/L (>0.3 mg/dL). The prevalence of RD defined as a GFR of <60 mL/min was 57%. During 6 months, WRF occurred in 161 (13.0%) patients. Predictors of WRF were vascular disease, the use of thiazide diuretics, and a baseline urea >9 mmol/L. Two hundred and sixty-three (21.6%) patients died, and baseline RD and WRF both predicted a higher mortality (P<0.001), whereas an improvement in renal function over the first 6 months predicted a lower mortality (hazard ratio 0.8, 95% confidence interval 0.6-1.0). CONCLUSION: In ambulatory patients with CHF, RD is common, commonly deteriorates over a relatively short period of time, is unlikely to recover substantially, and augurs a poor prognosis. PMID- 16364972 TI - Prognostic value of intra-left ventricular electromechanical asynchrony in patients with hypertrophic cardiomyopathy. AB - AIMS: We sought to assess the indexes of myocardial activation delay, using Doppler myocardial imaging (DMI), as potential predictors of cardiac events in patients with hypertrophic cardiomyopathy (HCM). The distribution and magnitude of left ventricular (LV) hypertrophy are not uniform in patients with HCM, which results in heterogeneity of regional LV systolic function. METHODS AND RESULTS: The study population included 123 HCM patients (39.4+/-5.9 years) and 123 age- and sex-matched healthy subjects, followed up for 48.4+/-8.8 months. By use of pulsed DMI, the following regional parameters were evaluated in six different basal myocardial segments: myocardial peak velocities and systolic time intervals; myocardial intraventricular (intra-V-Del) and interventricular (inter V-Del) systolic delays. DMI analysis in HCM showed lower myocardial systolic and early-diastolic peak velocities of all the segments. As for time intervals, HCM showed significant inter- and intra-V delays (P<0.0001), whereas homogeneous systolic activation of the ventricular walls was assessed in controls. During the follow-up, 16 cardiac deaths (12 sudden deaths) were observed in HCM patients. InHCM, DMI intra-V-Del was the most powerful independent predictor of sudden cardiac death (P<0.0001). In particular, an intra-V-Del>45 ms is identified with high sensitivity and specificity in HCM patients at higher risk of ventricular tachycardia and sudden cardiac death (test accuracy: 88.8%). CONCLUSION: In HCM patients, DMI indexes of intra-V-Del may provide additional information for selecting subgroups of HCM patients at increased risk of ventricular arrhythmias and sudden cardiac death at follow-up. Accordingly, such patients may be more actively identified for early intensive treatment and survey. PMID- 16364973 TI - Aspirin and clopidogrel resistance: an emerging clinical entity. AB - Antiplatelet therapy is a cornerstone of cardiovascular medicine. Aspirin and clopidogrel have emerged as critical therapies in the treatment of cardiovascular disease. Despite their efficacy, patients on these medications continue to suffer complications. Millions of patients are currently on low-dose antiplatelet therapy but it is unknown how many of these patients are under-treated or on the wrong medication. Aspirin and clopidogrel resistance are emerging clinical entities with potentially severe consequences such as recurrent myocardial infarction, stroke, or death. The mechanism of resistance remains incompletely defined, but there are specific clinical, cellular, and genetic factors that influence therapeutic failure. These factors range from physicians who fail to prescribe these medications despite appropriate indications to polymorphisms of platelet membrane glycoproteins. Rapid and accurate diagnosis of antiplatelet resistance also remains an issue as new bedside tests are developed. By understanding the mechanism of therapeutic failure and by improving the diagnosis of this clinical entity, a new era of individualized antiplatelet therapy may arise with routine measurements of platelet activity in the same way that cholesterol, blood pressure, and blood sugar are followed, thus improving the care for millions of people. PMID- 16364974 TI - Expression and transcriptional regulation of the GnRH receptor gene in human neuronal cells. AB - GnRH, acts via the GnRH receptor (GnRHR), plays a pivotal role in human reproduction by stimulating the synthesis and secretion of gonadotropins from pituitary gonadotropes. Studies have also suggested that it has other extra pituitary functions. To date, the transcriptional regulation of human GnRHR gene in the brain remains largely unknown. Recently, the human cerebellar medulloblastoma cell line TE-671 is found to express GnRH. We report here for the first time that GnRHR is also expressed in this neuronal cell line. Treatment with GnRHR agonist stimulated the phosphorylation of both ERK1/2 and JNK in the cells. Moreover, transient transfection of various human GnRHR promoter luciferase constructs into the cells identified an upstream promoter region located between -2197 and -1018. Important cis-acting regulatory elements were found at -1300/-1018 and -2197/- 1900, as deletion of either region caused a dramatic decrease in the promoter activity. An upstream GnRHR promoter element was identified to be important for basal transcription in the human neuronal TE 671 cells, in contrast to the previous finding that a downstream promoter is responsible for the gonadotrope-specific expression. Furthermore, we showed that antide (GnRHR antagonist) significantly stimulated the GnRHR promoter activity and inhibition of protein kinase C (PKC) pathway by staurosporine could also up regulate the promoter activity in dose- and time-dependent manners. Taken together, these data suggest that activation of the GnRHR by interacting with GnRH may transcriptionally down-regulate itself via the PKC pathway in human neuronal cells. PMID- 16364975 TI - Ikaros is expressed in human extravillous trophoblasts and involved in their migration and invasion. AB - The transcriptional factor Ikaros was originally found to function as a key regulator of lymphocyte differentiation. In addition, we have reported that Ikaros regulates the human placental leucine aminopeptidase (P-LAP)/insulin regulated aminopeptidase (IRAP) gene in choriocarcinoma trophoblastic cells, suggesting that Ikaros might be involved in placental development, while even its presence in human placenta remains undetermined. We therefore sought to clarify the location and roles of Ikaros in human placenta. Immunohistochemical analysis showed modest Ikaris expression in syncytium, and intense expression in extravillous trophoblasts (EVTs) in first trimester placenta. Western blot analysis showed that villous trophoblasts principally expressed Ikaros-2/3, while Ikaros-x (Ikx) was predominantly expressed in cultured EVTs. Furthermore, to investigate the functional role of Ikx in EVTs, the EVT cell line HTR-8/SVneo was infected with a retrovirus vector expressing the hemagglutinin (HA)-tagged dominant negative isoform Ikaros-6 (Ik6), which prevents the DNA-binding activity of Ikx. Antibody against HA showed successful transduction of Ik6 in HTR-8/SVneo cells on immunocytochemistry and Western blotting. Transduction of Ik6 significantly reduced the migratory and invasive abilities of HTR-8/SVneo cells. These results suggest that Ikx is involved in migration and invasion of EVTs in early placentation. PMID- 16364982 TI - The influence of gender on parasympathetic vasodilatation in the submandibular gland of the rat. AB - Parasympathetic vasodilatation in the rat submandibular gland is mediated by nitric oxide-dependent and -independent mechanisms (prostacyclin and endothelium derived hyperpolarizing factor (EDHF)). The purpose of this study was to determine the influence of gender on the relative contributions of each pathway to nerve-stimulated vasodilatation. Absolute increases in perfusion (laser Doppler flowmetry) were similar in male and female rats (in arbitrary perfusion units: 6159+/-4530 and 5601+/-3877 at 2 Hz; 15645+/-6830 and 14848+/-6118 at 5 Hz; and 22418+/-7660 and 18878+/-5864 at 10 Hz). However, expressed as a percentage increase above resting values, stimulated perfusion was higher in males than in females (P<0.05). In males both Nomega-nitro-L-arginine methyl ester (L-NAME) and indomethacin partly blocked parasympathetic vasodilatation at all frequencies tested (P<0.05). In female rats significant reductions in nerve stimulated perfusion were observed only at 2 and 5 Hz, but the effects of L-NAME were greater than in males (-64 compared with -45% at 2 Hz and -45 compared with 33% at 5 Hz, P<0.05). Indomethacin by itself had no apparent effect in females. The combined effects of L-NAME and indomethacin were dependent on the order of administration and on gender. Following L-NAME, indomethacin had no further effect in males or females. L-NAME reduced indomethacin-resistant vasodilatation in males and females, but the added effect of indomethacin was more pronounced in males. Finally, atropine-resistant vasodilatation was partly blocked by L-NAME, and the remaining vasodilatation was abolished by spantide I (substance P receptor antagonist). We conclude that NO, products of cyclo-oxygenase activity and EDHF all play a role in parasympathetic vasodilatation, but that NO and EDHF are the major endothelium-derived vasodilators in the rat submandibular gland. In addition, when other pathways are blocked EDHF makes a greater contribution in females. Lastly, both vasoactive intestinal peptide and substance P contribute to the atropine-resistant vasodilatation. PMID- 16364983 TI - Regulation of sexual behaviour in male macaques by sex steroid modulation of the serotonergic system. AB - The view that androgen action is the primary impetus underlying male-typical behaviour has been irrevocably altered by the profound perturbations in social and sexual behaviour observed in recent models of oestrogen insufficiency in male mice. Evidence is also accumulating for an involvement of oestrogens in the modulation of neural systems that are thought to play important roles in male reproductive functioning. Specifically, the serotonergic system is implicated in diverse autonomic functions, most or all of which are sensitive to oestradiol as well. Although their interaction domains have yet to be examined in male primates, roles have been established for both oestrogen and serotonin in the regulation of male sexual behaviour. We used a blinded, sham-treated and self controlled, randomized, multitreatment cross-over design to test the hypothesis that male sexual behaviour is regulated by oestrogen modulation of the serotonergic system in intact male Japanese macaques. Regression analysis revealed that oestradiol and whole blood tryptophan, but not testosterone or 5alpha-dihydrotestosterone, had additive, independent effects on male potentia over a range of hormone concentrations, whereas androgens were confirmed to be the primary determinants of sexual motivation. We suggest that modulation of the serotonergic system by 'female hormones' may be fundamental to the regulation of male mating success in higher primates. This might also explain, at least in part, why significant correlations between steroid hormones and male copulatory behaviour have traditionally proven so elusive in this order, thereby warranting a re-evaluation of the current notion that male sexual behaviour has been emancipated from activational hormonal control in higher primates. PMID- 16364984 TI - EBV-associated nasal-type NK/T-cell lymphoma of the nasal cavity/paranasal sinus in a renal allograft recipient. PMID- 16364985 TI - Persistent and symptomatic post-transplant hyperparathyroidism: a dramatic response to cinacalcet. PMID- 16364986 TI - Prevention of clot formation during haemodialysis using the direct thrombin inhibitor melagatran in patients with chronic uraemia. PMID- 16364987 TI - Risk factors and outcome of focal and segmental glomerulosclerosis recurrence in adult renal transplant recipients. AB - BACKGROUND: Recurrence of nephrotic syndrome (NS) after renal transplantation for primary focal segmental glomerulosclerosis (FSGS) is a frequent and still unpredictable complication. However, risk factors for recurrence have not yet been clearly identified. METHODS: Data from 33 patients who underwent 35 renal transplantations for FSGS in two French centres are reported. RESULTS: Recurrent NS occurred in 12 transplant recipients (34%). A significantly higher number of patients in the group with recurrence (R group) compared with the group without recurrence (NR group) received cyclosporine for FSGS treatment before transplantation (83.3% vs 43.4%, P<0.02). Donors of R group recipients were significantly older than those of the non-NR group recipients (42.8 years vs 35 years, P<0.05). A higher number of patients from the R group required post transplantation dialysis (33.3% vs 17.4%, P = 0.002). Surprisingly, acute rejection occurred more frequently in patients of the NR group compared with the R group, although the difference was not significant. Among the 12 patients with NS relapse, 9 were treated with plasmapheresis. Graft loss related to recurrence occurred in 6 cases. The 5-year graft survival was significantly lower in patients with recurrent NS compared with patients without recurrence (57% vs 82%, P<0.001). CONCLUSION: This study confirms the benefit to identify in the future clinical or biological predictive risk factors for NS recurrence after renal transplantation. It also indicates that donor age is a reliable risk factor for recurrence in adult recipients and suggests for the first time a possible opposite relationship between recurrent FSGS and acute rejection. PMID- 16364988 TI - Is oxidative stress implicated in high bone turnover in end-stage renal disease (ESRD)? PMID- 16364989 TI - Treatment of subcapsular haematoma, a complication of extracorporeal shock wave lithotripsy (ESWL), by percutaneous drainage. PMID- 16364990 TI - Evaluation of peritoneal transport properties at onset of peritoneal dialysis and longitudinal follow-up. AB - BACKGROUND: The clinical determinants of baseline peritoneal membrane (PM) transport characteristics, as evaluated by a hypertonic peritoneal equilibration test (PET), remain ill-defined. Likewise, the longitudinal evolution of PM transport properties in peritoneal dialysis (PD) patients given automated PD (APD) and icodextrin still needs to be determined precisely. The aims of the present study were (1) to determine the clinical and biological factors affecting PM transport characteristics at PD onset and (2) to assess the longitudinal evolution of these markers. METHODS: Seventy-two consecutive patients performed a baseline 3.86% glucose dialysate PET and were enrolled. Subgroups of 35 and 18 patients underwent another PET 1 and 2 year(s) later, respectively, and were included in the longitudinal part. For each patient, clinical and biological data were reviewed and PM transport markers calculated. RESULTS: At onset of PD, angiotensin-converting enzyme (ACE) inhibitor intake (r = 0.31, P = 0.01), presence of a diabetes (r = 0.26, P = 0.03) and body surface area (BSA) (r = 0.26, P = 0.03) independently affected the mass transfer area coefficient (MTAC) of creatinine. Serum albumin (r = -0.46, P<0.001) and net ultrafiltration (r = 0.33, P = 0.009) inversely correlated with MTAC creatinine. Sodium sieving was inversely correlated with BSA (r = -0.33, P = 0.01). Serum albumin also inversely correlated with albumin clearance (r = -0.39, P = 0.02). Finally, the independent covariates that affected alpha2-macroglobulin clearance were age (P = 0.03), diabetes (P = 0.01) and the level of residual renal function (P<0.01). Serum albumin decreased with time on PD (P = 0.02). A rise in small solute transport and a decrease in net ultrafiltration, but no change in protein clearances, were also observed after 2 years of PD. CONCLUSIONS: Transport properties across the PM, as evaluated by MTAC creatinine and sodium sieving determinations, are correlated with anthropometric characteristics (BSA) and by comorbid conditions (witnessed by the presence of diabetes, a low serum albumin concentration and the prescription of an ACE inhibitor). The short-term evolution (2 years) of the PM transport properties of patients on APD and icodextrin is still characterized by a progressive increase in small solute transport and a loss of ultrafiltration capacity, as documented in ancient studies, but not with a modification in protein clearances. This conclusion merits, however, to be further evaluated in a larger cohort of PD patients after a longer follow-up. PMID- 16364991 TI - A puzzling case of high serum creatinine in a healthy woman. PMID- 16364992 TI - Diameter measurements of the forearm cephalic vein prior to vascular access creation in end-stage renal disease patients: graduated pressure cuff versus tourniquet vessel dilatation. AB - BACKGROUND: Preoperative assessment of forearm superficial venous diameter may predict early failure of newly created arteriovenous fistulas for haemodialysis access. However, early failure and non-maturation rates remain high (up to 30%) and reported cut-off diameters are inconsistent. We hypothesize that this inconsistency is due to differences in the methods used to achieve venous dilatation prior to diameter measurements and daily variation in superficial venous diameter. We furthermore hypothesize that the use of a cuff will lead to a better inter-observer agreement since the applied pressure can be precisely determined. The purpose of this study was to determine inter-observer agreement of superficial venous diameter measurement under venous congestion by using either a graduated pressure cuff or tourniquet and furthermore, to determine daily variations in superficial venous diameter. METHODS: Diameter measurements were performed by two observers on days 1 and 3, in 21 end-stage renal disease patients using either a cuff (60 mmHg) or tourniquet. Measurements were carried out in random order and observers were blinded for each other's results. Inter observer agreement was expressed as interclass correlation coefficients. Variance components analysis was used to determine possible causes of disagreement. RESULTS: Using a cuff, mean venous diameter was 1.8 mm (range, 0.7-3.3 mm). When a tourniquet was used, the mean diameter was 1.8 mm (range, 0.6-3.2 mm). Interclass correlation coefficients between observers were 0.76 and 0.74 for the use of a cuff and tourniquet, respectively. Diameter measurements were revealed to be observer independent. Variations in venous diameter were determined by the patient and the interaction of patient and day. Repeated assessment of venous diameter on different days revealed a variation coefficient of 26.4% when using a cuff, and 26.5% when using a tourniquet. CONCLUSIONS: Venous diameter assessment is observer and congestion method independent. Daily variations in forearm superficial venous diameters should be taken into account when defining and using cut-off diameters prior to vascular access surgery. PMID- 16364993 TI - Autoinfarction of the parathyroid gland diagnosed by power Doppler ultrasonography in a patient with secondary hyperparathyroidism. PMID- 16364994 TI - Estimation of glomerular filtration rate in patients with normal serum creatinine undergoing primary PCI: is it really normal? PMID- 16364995 TI - Obituary: Richard Doll. PMID- 16364996 TI - Is it time to abandon the food frequency questionnaire? PMID- 16364997 TI - Role of lipid peroxidation in the epidemiology and prevention of breast cancer. AB - We have recently proposed a common mechanistic pathway by which obesity and hypertension lead to increased renal cell cancer risk. Our hypothesis posits lipid peroxidation, which is a principal mechanism in rodent renal carcinogenesis, as an intermediate step that leads to a final common pathway shared by numerous observed risks (including obesity, hypertension, smoking, oophorectomy/hysterectomy, parity, preeclampsia, diabetes, and analgesics) or protective factors (including oral contraceptive use and alcohol) for renal cell cancer [Cancer Causes Control 2002;13:287-93]. During this exercise, we have noticed how certain risk factors for renal cell carcinoma are protective for breast cancer and how certain protective factors for renal cell carcinoma increase risk for breast cancer. Parity and oophorectomy, for example, are positively associated with renal cell carcinoma but are negatively associated with breast cancer. Similarly, obesity and hypertension are positively associated with renal cell carcinoma, but obesity is negatively associated with breast cancer in premenopausal women and hypertension during pregnancy is negatively associated with breast cancer. Furthermore, alcohol intake, negatively associated with renal cell carcinoma, is also positively associated with breast cancer. We propose here the possibility that lipid peroxidation may represent a protective mechanism in breast cancer. Although this runs counter to the conventional view that lipid peroxidation is a process that is harmful and carcinogenic, we present here the chemical and biological rationale, based on epidemiologic and biochemical data, which may deserve further consideration and investigation. PMID- 16364998 TI - Inflammation and endometrial cancer: a hypothesis. AB - Endometrial cancer is the most common gynecologic malignancy in the United States. Substantial epidemiologic data implicate an imbalance of estrogens and progestogens in the etiology of this disease. We propose that inflammation also plays a role in endometrial cancer development. Emerging laboratory data suggest that elevated levels of prostaglandin E(2) may underlie the transformation of normal endometrium to neoplastic tissue and that in vitro nonsteroidal anti inflammatory drugs may inhibit endometrial cancer cell growth. In this review, we suggest that the risk factors for endometrial cancer--unopposed estrogens, anovulation, polycystic ovary syndrome, excessive menstruation, early menarche, and late menopause--may be viewed as factors increasing the exposure of the endometrium to inflammation, whereas pregnancy and smoking, two likely protective factors, have the opposite effect. Chronic inflammation can induce rapid cell division, increasing the possibility for replication error, ineffective DNA repair, and subsequent mutations. A proinflammatory milieu can also directly increase estrogen production. Hence, inflammation may work in conjunction with or in addition to estrogen exposure in the development of endometrial cancer. PMID- 16364999 TI - A randomized trial of an intervention to increase fruit and vegetable intake in curatively treated patients with early-stage head and neck cancer. AB - The leading cause of death in patients who have had curatively treated early stage head and neck cancer is a second primary cancer of the upper aerodigestive tract (lung, esophagus, larynx, pharynx, and oral cavity cancers). Low fruit and vegetable intake has been associated with increased risk of primary head and neck cancer and the available data suggest that increasing intake following diagnosis may reduce the risk of a second primary cancer. The goal of this study was to develop and test an easily administered intervention to increase fruit and vegetable intake in these patients following diagnosis and treatment. The 6-month intervention was based on the Stage of Change model. Seventy-five early-stage head and neck cancer patients were randomized to either the intervention group or to the "blinded" control group, with diet change data available on 65 patients. Fruit and vegetable intake, assessed using a food frequency questionnaire, and plasma carotenoid concentrations were measured at baseline and at the end of the study period. The change in self-reported intake of fruit and vegetables (servings per day) over the study period was significantly greater (P = 0.009) in the intervention group (n = 35; +2.1) compared with the control group (n = 30; +0.5). Total plasma carotenoids, a biomarker of fruit and vegetable intake, increased by 70 nmol/L in the intervention group as compared with a reduction of 42 nmol/L in the control group, a relative difference of 12% (nonsignificant). An intervention that can be delivered in a physician's office resulted in a significant increase in intake of fruit and vegetables in early-stage head and neck cancer patients. PMID- 16365000 TI - Factorial validity and invariance of a survey measuring psychosocial correlates of colorectal cancer screening among African Americans and Caucasians. AB - BACKGROUND: Psychosocial constructs are widely used to predict colorectal cancer screening and are targeted as intermediate outcomes in behavioral intervention studies. Reliable and valid instruments for measuring general colorectal cancer screening psychosocial constructs are needed; yet, few studies have conducted psychometric analyses. This study replicated a five-factor structure for 16 theory-based, general colorectal cancer screening items measuring salience and coherence, cancer worries, perceived susceptibility, response efficacy, and social influence. In addition, we examined factorial invariance across race and sex. METHODS: African American and Caucasian patients (n = 1,413) attending an urban, primary care clinic were included in this study. These individuals completed a baseline survey as part of a colorectal cancer screening intervention trial. Single and multigroup confirmatory factor analyses using maximum likelihood estimation were done. RESULTS: The five-factor general colorectal cancer screening model provided excellent fit and was invariant across race-sex subgroups. CONCLUSIONS: The findings of invariance across sex and race subgroups support the use of these scales to measure group differences. PMID- 16365001 TI - No evidence of false reassurance among women with an inconclusive BRCA1/2 genetic test result. AB - BACKGROUND: Little is known about how women who receive an inconclusive result from BRCA1/2 testing interpret their result. Clinical observations suggest that some of them may be falsely reassured and, consequently, may not adhere to recommended surveillance. The purpose of this study is to evaluate whether women with inconclusive BRCA1/2 test results are falsely reassured. METHODS: Participants were adult women with a family history suggestive of a germ-line mutation in either the BRCA1 or the BRCA2 gene who underwent genetic testing in the context of the interdisciplinary research program INHERIT BRCAs. Data were collected using self-administered questionnaires at genetic counseling and 1 month after result disclosure. Reassurance was assessed through indicators of cancer risk perception, cancer worry, relief following result disclosure, painfulness of the test result, and its effect on quality of life. RESULTS: Five hundred women (105 carriers, 140 noncarriers, and 255 inconclusive) were included in this analysis. Compared to noncarriers, women with inconclusive results had higher cancer risk perception, were more worried about cancer, were less relieved by their test result, and perceived their quality of life as being more adversely affected by it. CONCLUSION: The differences observed between noncarriers and women who received an inconclusive result run counter to the hypothesis that the latter are falsely reassured following BRCA1/2 testing. For clinicians, our findings show the value of taking precautions to fully explain to women that inconclusive results do not rule out the possibility that they still may face a higher risk of developing breast and/or ovarian cancer. PMID- 16365002 TI - Urinary thymidine dimer as a marker of total body burden of UV-inflicted DNA damage in humans. AB - High levels of DNA damage are induced in human skin following exposure to UV radiation. Cyclobutane thymidine dimer (T = T) is the most common of these lesions, which are enzymatically removed as oligonucleotides from DNA and further degraded before excretion in urine. Analysis of such repair products in the urine could serve as a biomarker of total body burden of UV exposure. The aim of this study was to examine the kinetics of T = T excretion following a single tanning session in a commercial solarium and to validate the method by delivering different doses. Ten individuals used the solarium for a total of 35 sessions of body tanning. Urine was collected before UV exposure and daily thereafter (up to 5 or 11 days) and T = T was analyzed using a very sensitive and quantitative (32)P-postlabeling technique combined with high-performance liquid chromatography. Following exposure, T = T levels increased dramatically and reached a peak 3 days later; afterwards, the T = T levels gradually decreased. The total amount of T = T excreted differed about 5-fold among subjects given an equal dose. A 50% excretion time was calculated using the excretion data for the first 5 days and it was found to be between 55 and 76 hours for different individuals. There was a good correlation between the amount of T = T excreted during days 1 to 5 and the delivered UV dose. Reducing exposure time to 50% lowered the amount of T = T to 47%; if half of the lamps were covered, T = T decreased to 44%. Our data show that urinary T = T could be a suitable noninvasive biomarker for UV exposure; a finding which could also be applicable to studies in children. PMID- 16365003 TI - Sun protection and the development of melanocytic nevi in children. AB - Childhood sun exposure causes nevi (and melanoma), but there is little evidence regarding the effectiveness of sun protection strategies on the number of nevi. We previously found that boys but not girls receiving a school-based sun protection program had fewer nevi on their backs than controls. Here, we investigated whether specific program components (encouraging children to stay indoors in the middle of the day during summer, to wear clothing while outdoors, and to use sunscreen) were associated with fewer nevi. An observational analysis was done on data from a sun protection trial in 1,623 children in Perth, Australia. The outcome was number of nevi on the back 6 years after baseline, when the children were 12 years old. Information on sun protection was obtained by questionnaires 4 and 6 years after baseline. The data were analyzed by mixed effects linear regression. The time spent outdoors between 11 a.m. and 2 p.m. and the proportion of total time outdoors that was between these hours were positively associated with number of nevi. Ratios of mean counts for doubling the respective measures were 1.09 [95% confidence interval (95% CI), 1.05-1.12] and 1.10 (95% CI, 1.05-1.14). Children whose backs were covered <70% of the time while outdoors had 1.53 (95% CI, 1.34-1.75) times more nevi than children whose backs were always covered. Using sunscreen on the back when it was uncovered was not associated with number of nevi (P = 0.59). Children who stayed indoors in the middle of the day and wore clothing while outdoors had fewer nevi. PMID- 16365004 TI - Associations of mammographic density with dietary factors in Japanese women. AB - BACKGROUND: A high percentage of mammographic dense area has been strongly associated with a risk of beast cancer. The present cross-sectional study evaluated the relations of percent density with dietary factors, such as fats, protein, dietary fiber, and soy isoflavones. METHODS: Study subjects were 601 (348 premenopausal and 253 postmenopausal) Japanese women who were recruited from a mammographic screening center. The size of the total breast area and the dense area were measured quantitatively using an automated mammographic mass detection method. Intakes of nutrients were estimated with a validated semiquantitative food-frequency questionnaire. RESULTS: The crude means of the percent density were 39.2% and 18.9% in premenopausal and postmenopausal women, respectively. There were no significant associations of any dietary factors with the percent density in premenopausal women. In postmenopausal women, percent density was significantly positively associated with intakes of protein, total fat, and saturated fat after controlling for covariates; the increase in the means of percent density were 7.2%, 5.6%, and 9.2% in the highest versus lowest quartile of intakes for protein, total fat, and saturated fat, respectively (P for linear trend were 0.006, 0.04, and 0.01, respectively). Carbohydrate intake was inversely associated with percent density; the mean of percent density was 6.0% lower in the highest versus the lowest quartile of intake (P(trend) = 0.03). The associations of dietary factors with dense area were very similar to those with percent density. CONCLUSION: These dietary factors may have implications for the risk of breast cancer in postmenopausal women. PMID- 16365005 TI - Relationship of obesity and physical activity with C-peptide, leptin, and insulin like growth factors in breast cancer survivors. AB - INTRODUCTION: Obese and physically inactive breast cancer patients may have poorer survival compared with lighter weight and more active women. Several obesity-related and physical activity-related hormones and peptides may explain this association, including insulin, leptin, insulin-like growth factor-I (IGF I), and IGF-binding protein-3. Few studies have examined the associations between obesity, physical activity, and these hormones/peptides among breast cancer survivors. PURPOSE: To determine whether obesity and physical activity are associated with insulin, IGFs, and leptin levels in a population-based sample of 710 women diagnosed with in situ to stage IIIA breast cancer and enrolled in the Health, Eating, Activity, and Lifestyle Study. METHODS: We collected a blood sample and information on physical activity among women diagnosed 2 to 3 years earlier using an interview-administered questionnaire. Trained staff measured weight. C-peptide, leptin, and IGFs were assayed by RIA. Mean hormone levels within body mass index and physical activity categories were adjusted for confounders using analysis of covariance methods. RESULTS: We observed higher C peptide (P for trend = 0.0001) and leptin (P for trend = 0.0001) levels and lower IGF-I levels (P for trend = 0.0001) with higher levels of body mass index. We observed lower C-peptide (P for trend = 0.001) and leptin (P for trend = 0.001) levels and higher IGF-I (P for trend = 0.0037) and IGF-binding protein-3 (P for trend = 0.055) levels with higher levels of physical activity. CONCLUSIONS: Increasing physical activity and decreasing body fat may be a reasonable intervention approach toward changing insulin and leptin, thereby potentially influencing breast cancer prognosis. PMID- 16365006 TI - Early life factors and incidence of proliferative benign breast disease. AB - Proliferative benign breast disease is a marker of increased breast cancer risk, yet little is known about its etiology. Most studies of benign breast disease have focused only on risk factors in adulthood, despite growing evidence that factors in early life influence breast cancer risk. We explored the relations of several early life factors with incidence of proliferative benign breast disease among 71,896 premenopausal women in the Nurses' Health Study II who recalled their body fatness at young ages, physical activity in adolescence, birthweight, and history of being breastfed. Between 1991 and 1997, 901 of these women were identified as having proliferative benign breast disease from a centralized pathology review. Relative risks (RR) and 95% confidence intervals (95% CI) were estimated from Cox proportional hazards models. Greater childhood body fatness (ages 5-10) was associated with decreased risk of proliferative benign breast disease; the multivariate RR (95% CI) for the most overweight compared with the most lean was 0.61 (0.44-0.86; P(trend) < 0.0001) and remained significant after adjustment for current body mass index. Body mass index at age 18 was also inversely associated with incidence of proliferative benign breast disease, with a multivariate RR (95% CI) of 0.67 (0.52-0.88) for those who were > or =25 kg/m(2) compared with those who were <19 kg/m(2) (P(trend) = 0.001). There were no clear associations for physical activity in adolescence, birthweight, or being breastfed. These results indicate that premenopausal women who were heavier at young ages have lower incidence of proliferative benign breast disease, consistent with previous findings for breast cancer. PMID- 16365007 TI - Dairy, calcium, and vitamin D intake and postmenopausal breast cancer risk in the Cancer Prevention Study II Nutrition Cohort. AB - BACKGROUND: Calcium, vitamin D, and dairy products are highly correlated factors, each with potential roles in breast carcinogenesis. Few prospective studies have examined these relationships in postmenopausal women. METHODS: Participants in the Cancer Prevention Study II Nutrition Cohort completed a detailed questionnaire on diet, vitamin and mineral supplement use, medical history, and lifestyle in 1992 to 1993. After exclusion of women with a history of cancer and incomplete dietary data, 68,567 postmenopausal women remained for analysis. During follow-up through August 31, 2001, we identified 2,855 incident cases of breast cancer. Multivariate-adjusted rate ratios (RR) were calculated using Cox proportional hazards models. RESULTS: Women with the highest intake of dietary calcium (>1,250 mg/d) were at a lower risk of breast cancer than those reporting < or =500 mg/d [RR, 0.80; 95% confidence interval (95% CI), 0.67-0.95; P(trend) = 0.02]; however, neither use of supplemental calcium nor vitamin D intake was associated with risk. Consumption starting at two or more servings of dairy products per day was likewise inversely associated with risk (RR, 0.81; 95% CI, 0.69-0.95; P(trend) = 0.002, compared with <0.5 servings/d). The associations were slightly stronger in women with estrogen receptor-positive tumors comparing highest to lowest intake: dietary calcium (RR, 0.67; 95% CI, 0.51-0.88; P(trend) = 0.004); dairy products (RR, 0.73; 95% CI, 0.57-0.93; P(trend) = 0.0003), and dietary vitamin D (RR, 0.74; 95% CI, 0.59-0.93; P(trend) = 0.006). CONCLUSIONS: Our results support the hypothesis that dietary calcium and/or some other components in dairy products may modestly reduce risk of postmenopausal breast cancer. The stronger inverse associations among estrogen receptor-positive tumors deserve further study. PMID- 16365008 TI - Migration history, acculturation, and breast cancer risk in Hispanic women. AB - BACKGROUND: Migrant studies have shown that breast cancer risk increases in women who move from countries with low incidence rates to countries with high rates. We examined the influence of migration history and acculturation on breast cancer risk in Hispanic women ages 35 to 79 years. METHODS: In a population-based case control study conducted in the San Francisco Bay Area, information on migration history, language usage, and other risk factors for breast cancer was collected through an in-person interview for 991 cases and 1,285 controls. RESULTS: Breast cancer risk was 50% lower in foreign-born Hispanics than U.S.-born Hispanics. Risk increased with increasing duration of residence in the United States, decreasing age at migration, and increasing acculturation. Among long-term foreign-born residents, risk was lower among Hispanics who moved to the United States at age > or =20 years and those who spoke mostly Spanish. The difference in risk between third-generation or higher-generation Hispanics and recent migrants from rural areas was approximately 6-fold in postmenopausal women and 4 fold in premenopausal women. Adjustment for differences in the distribution of breast cancer risk factors greatly attenuated the associations with migration patterns in premenopausal women; reduced risks remained only in those who resided in the United States for <10 years or migrated at age > or =30 years. In postmenopausal women, a 25% to 30% lower risk remained among long-term residents and those who migrated to the United States before age 20 years. CONCLUSIONS: These findings suggest the importance of yet unidentified protective factors among both recent premenopausal migrants and postmenopausal migrants. PMID- 16365009 TI - Regional differences in breast cancer survival despite common guidelines. AB - PURPOSE: Despite a uniform regional breast cancer care program, breast cancer survival differs within regions. We therefore examined breast cancer survival in relation to differences in diagnostic activity, tumor characteristics, and treatment in seven Swedish counties within a single health care region. METHODS: We conducted a population-based observational study using a clinical breast cancer register in one Swedish health care region. Eligible women (n = 7,656) ages 40 to 69 years diagnosed with primary breast cancer between 1992 and 2002 were followed up until 2003. The 7-year relative survival ratio was used to estimate breast cancer survival. Excess mortality was modeled using Poisson regression to study differences in survival between counties. RESULTS: The 7-year relative survival for breast cancer patients was significantly lower (up to 7% in absolute risk difference) in one county (county A) compared with the others. This difference existed only among women diagnosed before 1998, ages 50 to 59 years, and was strongest among stage II breast cancer patients. Adjustment for amount of diagnostic activity eliminated the survival differences among the counties. The amount of diagnostic activity was also lower in county A during the same time period. After county A, during 1997-1998, began to adhere strictly to the regional breast cancer care program, neither any survival differences nor diagnostic activity differences were observed. INTERPRETATIONS: Markers of diagnostic activity explained survival differences within our region, and the underlying mechanisms may be several. Low diagnostic activity may entail later diagnosis or inadequate characterization of the tumor and thereby missed treatment opportunities. Strengthening of multidisciplinary management of breast cancer can improve survival. PMID- 16365010 TI - AIB1 polymorphisms predict aggressive ovarian cancer phenotype. AB - PURPOSE: The androgen receptor (AR) harbors a polymorphic CAG repeat sequence in exon 1, coding for a polyglutamine tract whose length inversely correlates with AR transactivation function. AIB1, an AR coactivator, expresses a similar polymorphic glutamine sequence within the carboxyl-terminal coding region. We hypothesized that genotypic variations in the androgen-signaling pathway promote aggressive epithelial ovarian cancer biology, and sought to examine the effect of AIB1 genotype on clinical outcome. EXPERIMENTAL DESIGN: Genotype analysis of the AIB1 CAG repeat region was done on 89 patients with epithelial ovarian cancer. Medical records were reviewed for clinicopathologic factors and survival. Data were examined using the chi(2) test and Kaplan-Meier survival and Cox regression analyses. RESULTS: We identified four AIB1 genotypes, with glutamine codon lengths of 26, 28, 29, and 30. Patients with a short AIB1 genotype (with < or =28 CAG repeats) showed statistically shorter time to disease recurrence compared to those with a long genotype (> or =29 CAG repeats; 15.0 versus 30.0 months; P = 0.01). Patients with short AIB1 also showed decreased overall survival (57.0 months) compared to those with a long genotype (median survival not yet reached; P = 0.02). When controlling for established prognostic factors, multivariate analysis identified the presence of a short AIB1 genotype as an independent poor prognostic factor for overall survival (P = 0.05). CONCLUSIONS: These data suggest that short AIB1 genotypes may promote aggressive malignant phenotypes of epithelial ovarian cancer. PMID- 16365011 TI - Regular analgesic use and risk of endometrial cancer. AB - BACKGROUND: Analgesic use has been implicated in the chemoprevention of a number of solid tumors, but thus far, no previous research has focused on the role of aspirin in endometrial cancer etiology. METHODS: We conducted a hospital-based case-control study of 427 women with primary, incident endometrial cancer, and 427 age- and residence-matched controls without benign or malignant neoplasms. All participants received medical services at Roswell Park Cancer Institute in Buffalo, NY, and completed a comprehensive epidemiologic questionnaire. Women who reported analgesic use at least once a week for at least 6 months were classified as regular users and served as the reference group throughout the analyses. We used unconditional logistic regression analyses to compute crude and adjusted odds ratios (OR) with corresponding 95% confidence intervals (CI). RESULTS: Compared with nonusers, regular aspirin users were not at reduced risk of endometrial cancer (adjusted OR, 0.91; 95% CI, 0.66-1.26), nor were women with the highest frequency, duration, or cumulative lifetime aspirin use. When the sample was divided by body mass index status, regular aspirin use was not associated with risk among women classified as normal weight or overweight, but a significant risk reduction was seen for obese women (adjusted OR, 0.50; 95% CI, 0.27-0.92). Significant decreases in risk were also observed for obese women with the greatest frequency, duration, and cumulative aspirin use. No significant associations in the overall sample or among obese women were noted for acetaminophen use. CONCLUSION: We observed no evidence of an overall chemoprotective effect of aspirin on endometrial cancer risk, but the significant risk reductions among obese women warrant further investigation. PMID- 16365012 TI - Relationship of benign gynecologic diseases to subsequent risk of ovarian and uterine tumors. AB - OBJECTIVE: Although endometriosis and uterine leiomyomas are common conditions, the extent to which either is associated with certain types of malignancies remains uncertain. METHODS: Using record linkage techniques, we assessed the relationships between hospital and outpatient admissions for endometriosis or leiomyomas and the development of ovarian and uterine cancers in Denmark between 1978 and 1998. Based on a population-based cohort exceeding 99,000 women, including 2,491 ovarian cancers, 860 borderline ovarian tumors, and 1,398 uterine cancers, we derived relative risks (RR) and 95% confidence intervals (95% CI) associated with overall and histology-specific tumor risks after adjustment for calendar time and reproductive characteristics. RESULTS: Endometriosis seemed to predispose to the development of ovarian cancer, with the association restricted to endometrioid or clear cell malignancies. Five or more years after the diagnosis of endometriosis, the RRs (95% CIs) were 2.53 (1.19-5.38) for endometrioid (7 exposed cases) and 3.37 (1.24-9.14) for clear cell (4 exposed cases) malignancies. Uterine leiomyomas were associated with increases in the risk of uterine malignancies, particularly sarcomas, where the RRs (95% CIs) were 20.80 (11.32-38.22) for women with 1 to 4 years of follow-up (11 exposed cases) and 5.70 (2.27-14.32) for those with more extended follow-up (5 exposed cases). CONCLUSION: In combination with clinical, pathologic, and molecular data, our results support that some endometriotic lesions may predispose to clear cell and endometrioid ovarian cancers. Uterine leiomyomas also showed a strong connection with subsequent uterine sarcomas, although it was difficult to decipher whether this reflected detection bias, shared risk factors, or an etiologic relationship. PMID- 16365013 TI - Associations between ERalpha, ERbeta, and AR genotypes and colon and rectal cancer. AB - Estrogen and androgens are thought to be involved in the etiology of colorectal cancer. We evaluate genetic variants of the estrogen receptor genes (ERalpha and ERbeta) and the androgen receptor gene (AR). We use data from two large case control studies of colon (n = 1,580 cases and 1,968 controls) and rectal (n = 797 cases and 1,016 controls) cancer. We evaluated the 351A >G XbaI polymorphism of ERalpha, the 1,082 G >A and CA repeat polymorphisms of ERbeta, and the CAG repeat of AR. Having two 25 or more CA repeats in ERbeta was associated with an increased relative risk of colon cancer in women [odds ratio (OR), 2.13; 95% confidence interval (95% CI), 1.24-3.64] but not in men (P(interaction) relative excess risk from interaction < 0.01; multiplicative = 0.03). Increasing number of AR CAG repeats was directly associated with colon cancer among men (OR, 1.28; 95% CI, 1.06-1.54), but not women (OR, 0.83; 95% CI, 0.68-1.02); the interaction P value for AR gene x sex was <0.01. Taking hormone replacement therapy (HRT) was associated with a reduced risk of colon cancer in the presence of the R allele of the ERbeta gene, whereas an R allele was associated with increased risk among postmenopausal women who did not take HRT. Postmenopausal women not using HRT who had > or =25 CA repeats of the ERbeta gene had over a 6-fold increased risk of colon cancer (OR, 6.71; 95% CI, 2.89-15.6). Our results suggest that the ERbeta gene is more important than ERalpha in the etiology of colorectal cancer. PMID- 16365014 TI - GSTP1 and GSTA1 polymorphisms interact with cruciferous vegetable intake in colorectal adenoma risk. AB - The possible interplay between cruciferous vegetable consumption, functional genetic variations in glutathione S-transferases (GST) M1, T1, P1, and A1, and colorectal adenomas, was investigated in a Dutch case-control study. The GSTM1 and GSTT1 deletion polymorphisms, and the single nucleotide polymorphisms in GSTP1 (A313G) and in GSTA1 (C-69T) were assessed among 746 cases who developed colorectal adenomas and 698 endoscopy-based controls without any type of colorectal polyps. High and low cruciferous vegetable consumption was defined based on a median split in the control group. High consumption was slightly positively associated with colorectal adenomas [odds ratio (OR) 1.15; 95% confidence interval, 0.92-1.44]. For GSTP1, a positive association with higher cruciferous vegetable intake was only apparent in individuals with the low activity GSTP1 genotype (GG genotype, OR 1.94; 95% confidence interval, 1.02 3.69). This interaction was more pronounced in men, with higher age and with higher meat intake. The GSTA1 polymorphism may have a modifying role as well: the OR for higher intake compared with lower intake was 1.57 (0.93-2.65) for individuals homozygous for the low expression variant (TT genotype). This seemed to be stronger with younger age and higher red meat intake. Cruciferous vegetable consumption and the combined GSTA1 and GSTP1 genotypes showed a statistically significant interaction (P = 0.034). The GSTM1 and GSTT1 genotypes did not seem to modify the association between cruciferous vegetable intake and colorectal adenomas. In conclusion, GSTP1 and GSTA1 genotypes might modulate the association between cruciferous vegetable intake and colorectal adenomas. PMID- 16365015 TI - Human papillomavirus 16, 18, and 33 infections and risk of prostate cancer: a Nordic nested case-control study. AB - Epidemiologic evidence of sexual history has emerged as a consistently found risk factor for prostate cancer. Some studies have reported an association between human papillomavirus (HPV) infections and prostate cancer. We did a nested case control study within cohorts of more than 200,000 men enrolled in three Nordic biobanking projects. Follow-up using cancer registry linkages identified 804 prospectively occurring prostate cancer cases. Four control subjects per case were randomly selected from eligible sets of matched subjects that were alive and free of cancer at the time of diagnosis of the corresponding case and were matched to cases on biobank cohort, age (+/-2 years), county of residence, and date of blood sampling (+/-2 months in the Finnish and Swedish cohorts, +/-6 months in the Norwegian cohort). The serum samples were analyzed by standard ELISAs for the presence of immunoglobulin G antibodies against HPV types 16, 18, and 33. The joint HPV-16/HPV-18/HPV-33 seroprevalence in the joint cohort was 13.4% (107 of 799) among cases and 14.0% (363 of 2,596) among controls (odds ratio, 0.94; 95% confidence interval, 0.74-1.19). There were no noteworthy differences when the data were analyzed by different HPV type, country, or antibody levels. Our data do not support an association between serologic markers of HPV-16, HPV-18, and HPV-33 infections and risk of prostate cancer. PMID- 16365016 TI - Quantitative analysis of plasma TP53 249Ser-mutated DNA by electrospray ionization mass spectrometry. AB - A mutation in codon 249 of the TP53 gene (249(Ser)), related to aflatoxin B(1) exposure, has previously been associated with hepatocellular carcinoma risk. Using a novel internal standard plasmid, plasma concentrations of 249(Ser) mutated DNA were quantified by electrospray ionization mass spectrometry in 89 hepatocellular carcinoma cases, 42 cirrhotic patients, and 131 nonliver diseased control subjects, all from highly aflatoxin-exposed regions of The Gambia. The hepatocellular carcinoma cases had higher median plasma concentrations of 249(Ser) (2,800 copies/mL; interquartile range: 500-11,000) compared with either cirrhotic (500 copies/mL; interquartile range: 500-2,600) or control subjects (500 copies/mL; interquartile range: 500-2,000; P < 0.05). About half (52%) of the hepatocellular carcinoma cases had >2,500 copies of 249(Ser)/mL plasma, corresponding to the prevalence of this mutation in liver tumors in The Gambia. In comparison, only 15% of control group and 26% of cirrhotic participants exceeded this level (P < 0.05). Further subset analysis revealed a statistically significant, quantitative relation between diagnosis of hepatocellular carcinoma and levels of 249(Ser) detected at 2,501 to 10,000 copies/mL plasma (odds ratio, 3.8; 95% confidence interval, 1.3-10.9) and at >10,000 copies/mL plasma (odds ratio, 62; 95% confidence interval, 4.7-820) when compared with control subjects and after adjusting for age, gender, recruitment site, hepatitis B and C serologic status, and total DNA concentration. Levels of >10,000 copies of 249(Ser)/mL plasma were also significantly associated with the diagnosis of hepatocellular carcinoma (odds ratio, 15; 95% confidence interval, 1.6-140) when compared with cirrhotic patients. Potential applications for the quantification of 249(Ser) DNA in plasma include estimation of long-term, cumulative aflatoxin exposure and selection of appropriate high-risk individuals for targeted intervention. PMID- 16365017 TI - Relationships between cigarette consumption and biomarkers of tobacco toxin exposure. AB - Epidemiologic studies show a dose-response relationship between cigarettes per day and health outcomes such as heart and lung disease, and health outcomes are related to some biomarkers of tobacco exposure. The objective of this study was to examine the relationships between cigarettes per day and levels of selected biomarkers of tobacco toxin exposure: carbon monoxide (CO), metabolites of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and polycyclic aromatic hydrocarbons [total 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanol (NNAL) and 1-hydroxypyrene (1-HOP), respectively], and total cotinine (cotinine plus cotinine-N-glucuronide). We did a cross-sectional analysis of merged data from (a) two clinical trials and (b) two cohorts of light smokers (total n = 400). The mean age of participants was 50.4 years and the range of cigarette consumption was 1 to 100/d; however, few subjects smoked >45 cigarettes/d (n = 12). Results show that levels of the biomarkers CO, total NNAL, and total cotinine increase with an increase in the number of cigarettes smoked per day, but not in a linear fashion. 1-HOP is a less discriminating biomarker as levels are relatively stable regardless of the number of cigarettes smoked per day. There is considerable variability in toxin measurement, especially at high levels of smoking. There was a significant correlation between cigarettes per day and total NNAL, 1-HOP, total cotinine, and CO. Total NNAL was highly significantly correlated with total cotinine and CO and also significantly correlated with 1-HOP. These findings suggest that the number of cigarettes smoked per day is not necessarily a reliable measure of toxin exposure and may underestimate tobacco toxin exposure at low levels of smoking or overestimate exposure at high levels of smoking. PMID- 16365018 TI - Longitudinal study of urinary phenanthrene metabolite ratios: effect of smoking on the diol epoxide pathway. AB - We have proposed that urinary phenanthrene metabolites could be used in a carcinogen metabolite phenotyping approach to identify individuals who may be susceptible to cancer induction by polycyclic aromatic hydrocarbons (PAH). In support of this proposal, we have developed methods for quantitation of r-1,t 2,3,c-4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene (PheT) and phenanthrols (HOPhe) in human urine. PheT is the end product of the diol epoxide metabolic activation pathway of PAH, whereas HOPhe are considered as detoxification products. In this study, we investigated the longitudinal consistency of these metabolites over time in smokers and nonsmokers and compared their levels. Twelve smokers and 10 nonsmokers provided urine samples daily for 7 days, then weekly for 6 weeks. Levels of PheT, HOPhe, and PheT/HOPhe ratios were relatively constant in most individuals, with mean coefficients of variation ranging from 29.3% to 45.7%. There were no significant changes over time in levels of the metabolites or in ratios. These results indicate that a single urine sample should be sufficient when comparing phenanthrene metabolites in different groups. PheT/HOPhe ratios were significantly higher in smokers than in nonsmokers, showing that smoking induces the diol epoxide metabolic activation pathway of phenanthrene. This finding is consistent with previous studies indicating that inducibility of PAH metabolism contributes to cancer risk in smokers. PMID- 16365019 TI - Active and passive smoking and lifestyle determinants of 8-oxo-7,8-dihydro-2' deoxyguanosine levels in human leukocyte DNA. AB - We investigated the effects of smoking and exposure to environmental tobacco smoke (ETS) on oxidative DNA damage by measuring 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo) levels in DNA of leukocytes of healthy donors (30 smokers, 29 nonsmokers, and 28 ETS-exposed subjects). Nonsmokers had lower 8 oxodGuo levels compared with smokers (5.94 +/- 0.87 x 10(-6) and 19.85 +/- 4.75 x 10(-6) 2-deoxyguanosine, respectively, means +/- SE, P = 0.00007). Subjects exposed to ETS had higher mean value of 8-oxodGuo compared with nonsmokers (9.18 +/- 1.53 x 10(-6) 2-deoxyguanosine, mean +/- SE), nonsignificant by univariate analysis (P = 0.074). Multiregression analysis indicated that the increase of 8 oxodGuo levels induced by ETS was significant (P = 0.045) and that coffee and tea consumption reduced DNA oxidation (P = 0.0053). Oxidative leukocyte DNA damage was positively correlated with plasma cotinine levels in ETS-exposed subjects (r = 0.47, P < 0.01, n = 28) and was increased by age in nonsmokers and ETS-exposed subjects (P = 0.049). The results seem to confirm that ETS exposure is capable of inducing some oxidative DNA damage in circulating leukocytes and that coffee and tea consumption might partially protect against smoking-induced oxidation damage. PMID- 16365020 TI - Interest in testing for genetic susceptibility to lung cancer among Black college students "at risk" of becoming cigarette smokers. AB - Receptivity to genetic testing for lung cancer susceptibility was assessed among African American college freshmen, who held attitudes favorable towards or had experimented with cigarette smoking. Students (n = 95) completed a telephone survey that assessed beliefs about genetics and lung cancer risk, interest in genetic testing, and expectations about the test outcome. Interest in being tested was moderately high (mean, 5; SD, 2.2; scale of 1-7) and highest among those who believed lung cancer was influenced by genetics (r = 0.22, P < 0.05) and those who expected to be at high risk (r = 0.27, P < 0.05). Overall, 34% thought if tested, the result would show high risk for lung cancer. In multivariate analyses, students' test result expectation was the only significant predictor of interest in testing. Those who believed the test would show them to be at higher risk were thrice more likely to be interested in testing than those who thought the test would show that they were at lower risk (odds ratio, 2.99; confidence interval, 1.03-8.64; P = 0.04). Future research is needed to understand how young adults will respond to genetic susceptibility feedback that confirms or contradicts their expectations about personal risks of smoking. PMID- 16365021 TI - Risk factors associated with the development of intestinal metaplasia in first degree relatives of gastric cancer patients. AB - Family relatives of gastric cancer patients have a higher risk of gastric cancer and premalignant gastric lesions. We sought to determine the risk factors associated with the presence of intestinal metaplasia in a large cohort of gastric cancer relatives. First-degree relatives of gastric cancer patients were invited for screening gastroscopy. Endoscopic gastric biopsies were obtained from the antrum and corpus. Gastric biopsies were analyzed for Helicobacter pylori infection, severity of inflammation, and presence of intestinal metaplasia. Stepwise logistic regressions were used to identify for risk factors associated with presence of intestinal metaplasia in cancer relatives. Two hundred seventy cancer relatives underwent screening endoscopy (median age, 42; 47% male and 48% siblings). Among them, 161 (59.6%) were H. pylori positive and 81 (30%) had confirmed intestinal metaplasia. The following factors were found to be associated with the presence of intestinal metaplasia: age, male sex, H. pylori infection, birth order, alcohol use, siblings with stomach cancer, childhood living conditions, and water supply. Individuals with intestinal metaplasia had more severe acute and chronic inflammation in the antrum and corpus (P < 0.003). With multiple logistic regression, H. pylori infection [odds ratio (OR), 3.23], male gender (OR, 2.09), age (OR, 1.07), and a history of gastric cancer in siblings (OR, 1.91) were independent factors associated with the development of intestinal metaplasia in cancer relatives. In conclusion, we have identified risk factors associated with gastric intestinal metaplasia in stomach cancer relatives, which may be useful in the understanding of gastric carcinogenesis in these high-risk individuals. PMID- 16365022 TI - The angiotensin I-converting enzyme gene insertion/deletion polymorphism is linked to early gastric cancer. AB - The insertion/deletion polymorphism of the angiotensin I-converting enzyme (ACE) gene has recently been linked to the pathogenesis and progression of human cancers. Using genomic DNA from 88 patients with early gastric cancer confined either to mucosa (pT(1a)) or submucosa (pT(1b)), we assessed the insertion (I) and deletion (D) polymorphism by PCR analysis and compared it with a large noncancer control population (n = 145). In the noncancer control group, the II genotype was observed in 33 (23%) individuals, whereas the ID and DD genotypes were found in 72 (50%) and 40 (27%) individuals, respectively. Interestingly, in the cancer group, we found the II genotype in six (7%) patients and the ID genotype in 46 (52%) individuals, whereas the DD genotype was observed in 36 (41%) individuals (P = 0.0034). Accordingly, the odds ratio for the II genotype was 0.20 [95% confidence interval (95% CI), 0.08-0.54; P = 0.009] and 0.55 for the ID/II genotype (95% CI, 0.31-0.96; P = 0.044) using the high-activity genotype DD as the reference category. No correlation was found among tumor type, tumor stage, the presence of Helicobacter pylori, and the ACE genotype. Our study provides further evidence that the ACE insertion/deletion gene polymorphism may be linked to the development of early gastric cancer. PMID- 16365023 TI - Androgen receptor and prostate-specific antigen gene polymorphisms and breast cancer in African-American women. AB - Several previous studies have found the CAG repeat polymorphism in exon 1 of the androgen receptor (AR) gene to be associated with breast cancer risk among some groups of Caucasian and Asian women. In a population-based case-control study of 488 African-American women (239 cases and 249 controls), we examined this polymorphism along with a polymorphism (-158 G/A) in an androgen-regulated gene (PSA) whose expression has been correlated with breast cancer prognosis. Overall, we did not observe any significant association between the CAG repeat polymorphism and breast cancer risk. However, among women with a first-degree family history of breast cancer, longer CAG repeats were associated with a significantly increased risk. Women carrying at least one longer allele [(CAG)n > or = 22] had a 3-fold increased risk compared to those with two shorter alleles (odds ratio, 3.18; 95% confidence interval, 1.08-9.36). There was no significant association between the PSA gene polymorphism and breast cancer risk, nor was there significant gene-gene interaction. In summary, our results further support that shorter CAG repeats (stronger AR transactivation activity) may reduce the risk of breast cancer, at least among some groups of women. Our data, however, are unable to provide evidence that PSA is the pathway through which the protective effect of androgens operates. PMID- 16365024 TI - Nipple fluid basic fibroblast growth factor in patients with breast cancer. AB - PURPOSE: It has been shown that early detection of breast cancer could save lives. Recently, there has been increasing interest in nipple fluid as a potential supplemental avenue for breast cancer diagnosis. EXPERIMENTAL DESIGN: In this study, we determined the levels of an angiogenic factor basic fibroblast growth factor (bFGF) in the nipple fluid of healthy subjects as well as patients with benign breast conditions, those at high risk for breast cancer, and patients with active breast cancer. ELISAs were used to measure bFGF. RESULTS: Nipple fluid bFGF levels were as follows (mean +/- SE): 158 +/- 17 pg/mL from benign breasts, 561 +/- 277 pg/mL from high-risk breasts, and 1,343 +/- 441 pg/mL from cancerous breasts. One-way ANOVA showed that the bFGF levels from cancerous breasts were significantly higher than those from benign and high-risk breasts (P = 0.0001 and P = 0.0193, respectively). After logarithmic transformation was applied to the data, high-risk breast bFGF levels were higher than those from benign breasts (P = 0.0028). With a cutoff level of 250 pg/mL, the sensitivity was 79.2%, specificity was 82.5%, and correct diagnosis was 66.4%. The area under the receiver operating characteristic curve was 0.86. CONCLUSIONS: We conclude that nipple fluid bFGF levels are progressively elevated in high-risk and cancerous breasts compared with benign breasts. The sensitivity and specificity of this test are promising compared with current breast cancer screening methods, and this test deserves further studies with larger clinical trials. Potential areas of usefulness include the detection of breast cancer risk or breast cancer, as well as the monitoring and/or prediction of the antiangiogenic effect of preventive therapies. PMID- 16365025 TI - Risk of non-Hodgkin lymphoma associated with polymorphisms in folate-metabolizing genes. AB - Genetic instability, including chromosomal imbalance, is important in the pathogenesis of lymphoproliferative disorders such as non-Hodgkin lymphoma (NHL). DNA synthesis and methylation, which are closely linked to folate metabolism and transport, may be affected by polymorphisms in genes involved in these pathways. Folate metabolism polymorphisms have been linked to acute lymphoblastic leukemia and colorectal cancer. To evaluate whether genetic variation in folate metabolism and transport may have a role in determining the risk of developing NHL, we analyzed several polymorphisms using DNA obtained as part of a large U.K. population-based case-control study of lymphoma. Polymorphisms studied include methylenetetrahydrofolate reductase (MTHFR) 677 C >T and 1298 A >C, methionine synthase (MTR) 2756 A>G, serine hydroxymethyltransferase (SHMT1) 1420 C >T, thymidylate synthase (TYMS) 1494del6 and 28-bp repeat, and reduced folate carrier (RFC) 80 G >A. Increased risks for NHL [odds ratio (OR), 1.48; 95% confidence intervals (CI), 1.12-1.97], and marginal zone lymphoma (OR, 3.38; 95% CI, 1.30 8.82) were associated with the TYMS 2R/3R variant. Marginal increased risks were also observed for diffuse large B cell lymphoma with the TYMS homozygous 6 bp deletion (OR, 1.61; 95% CI, 0.99-2.60) and for follicular lymphoma with RFC 80AA (OR, 1.44; 95% CI, 0.94-2.22) and TYMS 28-bp repeat 2R/3R (OR, 1.45; 95% CI, 0.96 2.2). We observed no association between NHL and haplotypes for MTHFR or TYMS. These findings are somewhat inconsistent with those of others, but may reflect differences in circulating folate levels between study populations. Thus, further investigations are warranted in larger series with dietary information to determine the roles that genetics and folic acid status play in the etiology of lymphoma. PMID- 16365026 TI - Data-dredging gene-dose analyses in association studies: biases and their corrections. AB - To examine the joint effect of multiple loci on disease risk, many case-control association studies used "gene-dose analyses." However, some researchers defined high-risk genotypes (or alleles) as those that have higher genotypic (allelic) frequencies in the case group compared with the control group in the study. This will lead to the total number of the "high-risk" genotypes (alleles) tending to be higher for the cases than for the controls as well, even if none of the studied loci were related to the disease. Monte-Carlo simulations done in this study showed that such a "data-dredging" gene-dose analysis could produce grossly biased results. A permutation correction method was proposed which could correct the biases very effectively. PMID- 16365027 TI - No association between residential exposure to petrochemicals and brain tumor risk. PMID- 16365028 TI - Hemoglobin A1c concentrations and risk of colorectal cancer in women. PMID- 16365029 TI - Cyclooxygenase 2 polymorphism (Val511Ala), nonsteroidal anti-inflammatory drug use and breast cancer in African American women. PMID- 16365030 TI - One-carbon metabolism and breast cancer risk: no association of MTHFR, MTR, and TYMS polymorphisms in the GENICA study from Germany. PMID- 16365031 TI - Insulin-like growth factor-I and mammographic breast density. PMID- 16365032 TI - Probing the functional link between androgen receptor coactivator and ligand binding sites in prostate cancer and androgen insensitivity. AB - The androgen receptor (AR) is a ligand-activated transcription factor required for male sex development and virilization and contributes to prostate cancer initiation and progression. High affinity androgen binding triggers conformational changes required for AR transactivation. Here we characterized naturally occurring AR gene mutations in the region of activation function 2 (AF2) that decrease or increase AR transcriptional activity by altering the region bounded by AF2 and the ligand binding pocket without affecting equilibrium androgen binding affinity. In the androgen insensitivity syndrome, germ line AR mutations increase the androgen dissociation rate and reduce AR FXXLF motif binding and the recruitment of steroid receptor coactivator (SRC)/p160 coactivator LXXLL motifs. In prostate cancer, somatic AR mutations in AF2 or near the bound ligand slow androgen dissociation and increase AR stabilization and coactivator recruitment. Crystal structures of the AR ligand binding domain bound to R1881 and FXXLF or LXXLL motif peptide indicate the mutations are proximal to the AF2 bound peptide, adjacent to the ligand pocket, or in a putative ligand gateway. The results suggest a bidirectional structural relay between bound ligand and coactivator that establishes AR functional potency in vivo. PMID- 16365033 TI - Solution structure and novel insights into the determinants of the receptor specificity of human relaxin-3. AB - Relaxin-3 is the most recently discovered member of the relaxin family of peptide hormones. In contrast to relaxin-1 and -2, whose main functions are associated with pregnancy, relaxin-3 is involved in neuropeptide signaling in the brain. Here, we report the solution structure of human relaxin-3, the first structure of a relaxin family member to be solved by NMR methods. Overall, relaxin-3 adopts an insulin-like fold, but the structure differs crucially from the crystal structure of human relaxin-2 near the B-chain terminus. In particular, the B-chain C terminus folds back, allowing Trp(B27) to interact with the hydrophobic core. This interaction partly blocks the conserved RXXXRXXI motif identified as a determinant for the interaction with the relaxin receptor LGR7 and may account for the lower affinity of relaxin-3 relative to relaxin for this receptor. This structural feature is likely important for the activation of its endogenous receptor, GPCR135. PMID- 16365034 TI - AIFsh, a novel apoptosis-inducing factor (AIF) pro-apoptotic isoform with potential pathological relevance in human cancer. AB - AIF is a main mediator of caspase-independent cell death. It is encoded by a single gene located on chromosome X, region q25-26 and A6 in humans and mice, respectively. Previous studies established that AIF codes for two isoforms of the protein, AIF and AIF-exB. Here, we identify a third AIF isoform resulting from an alternate transcriptional start site located at intron 9 of AIF. The resulting mRNA encodes a cytosolic protein that corresponds to the C-terminal domain of AIF (amino acids 353-613). We named this new isoform AIFshort (AIFsh). AIFsh overexpression in HeLa cells results in nuclear translocation and caspase independent cell death. Once in the nucleus, AIFsh provokes the same effects than AIF, namely chromatin condensation and large scale (50 kb) DNA fragmentation. In contrast, these apoptogenic effects are not precluded by the AIF-inhibiting protein Hsp70. These findings identify AIFsh as a new pro-apoptotic isoform of AIF, and also reveal that the first N-terminal 352 amino acids of AIF are not required for its apoptotic activity. In addition, we demonstrate that AIFsh is strongly down-regulated in tumor cells derived from kidney, vulva, skin, thyroid, and pancreas, whereas, gamma-irradiation treatment provokes AIFsh up-regulation. Overall, our results identify a novel member of the AIF-dependent pathway and shed new light on the role of caspase-independent cell death in tumor formation/suppression. PMID- 16365035 TI - Differential inhibition of Arabidopsis methionine adenosyltransferases by protein S-nitrosylation. AB - In animals, protein S-nitrosylation, the covalent attachment of NO to the thiol group of cysteine residues, is an intensively investigated posttranslational modification, which regulates many different processes. A growing body of evidence suggests that this type of redox-based regulation mechanism plays a pivotal role in plants, too. Here we report the molecular mechanism for S nitrosylation of methionine adenosyltransferase (MAT) of Arabidopsis thaliana, thereby presenting the first detailed characterization of S-nitrosylation in plants. We cloned three MAT isoforms of Arabidopsis and tested the effect of NO on the activity of the purified, recombinant proteins. Our data showed that incubation with GSNO resulted in blunt, reversible inhibition of MAT1, whereas MAT2 and MAT3 were not significantly affected. Cys-114 of MAT1 was identified as the most promising target of NO-induced inhibition of MAT1, because this residue is absent in MAT2 and MAT3. Structural analysis of MAT1 revealed that Cys-114 is located nearby the putative substrate binding site of this enzyme. Furthermore, Cys-114 is flanked by S-nitrosylation-promoting amino acids. The inhibitory effect of GSNO was drastically reduced when Cys-114 of MAT1 was replaced by arginine, and mass spectrometric analyses of Cys-114-containing peptides obtained after chymotryptic digestion demonstrated that Cys-114 of MAT1 is indeed S nitrosylated. Because MAT catalyzes the synthesis of the ethylene precursor S adenosylmethionine and NO is known to influence ethylene production in plants, this enzyme probably mediates the cross-talk between ethylene and NO signaling. PMID- 16365036 TI - Pyrithiamine as a substrate for thiamine pyrophosphokinase. AB - Thiamine pyrophosphokinase transfers a pyrophosphate group from a nucleoside triphosphate, such as ATP, to the hydroxyl group of thiamine to produce thiamine pyrophosphate. Deficiencies in thiamine can result in the development of the neurological disorder Wernicke-Korsakoff Syndrome as well as the potentially fatal cardiovascular disease wet beriberi. Pyrithiamine is an inhibitor of thiamine metabolism that induces neurological symptoms similar to that of Wernicke-Korsakoff Syndrome in animals. However, the mechanism by which pyrithiamine interferes with cellular thiamine phosphoester homeostasis is not entirely clear. We used kinetic assays coupled with mass spectrometry of the reaction products and x-ray crystallography of an equilibrium reaction mixture of thiamine pyrophosphokinase, pyrithiamine, and Mg2+/ATP to elucidate the mechanism by which pyrithiamine inhibits the enzymatic production of thiamine pyrophosphate. Three lines of evidence support the ability of thiamine pyrophosphokinase to form pyrithiamine pyrophosphate. First, a coupled enzyme assay clearly demonstrated the ability of thiamine pyrophosphokinase to produce AMP when pyrithiamine was used as substrate. Second, an analysis of the reaction mixture by mass spectrometry directly identified pyrithiamine pyrophosphate in the reaction mixture. Last, the structure of thiamine pyrophosphokinase crystallized from an equilibrium substrate/product mixture shows clear electron density for pyrithiamine pyrophosphate bound in the enzyme active site. This structure also provides the first clear picture of the binding pocket for the nucleoside triphosphate and permits the first detailed understanding of the catalytic requirements for catalysis in this enzyme. PMID- 16365037 TI - Regulation of de novo purine biosynthesis by methenyltetrahydrofolate synthetase in neuroblastoma. AB - 5-Formyltetrahydrofolate (5-formylTHF) is the only folate derivative that does not serve as a cofactor in folate-dependent one-carbon metabolism. Two metabolic roles have been ascribed to this folate derivative. It has been proposed to 1) serve as a storage form of folate because it is chemically stable and accumulates in seeds and spores and 2) regulate folate-dependent one-carbon metabolism by inhibiting folate-dependent enzymes, specifically targeting folate-dependent de novo purine biosynthesis. Methenyltetrahydrofolate synthetase (MTHFS) is the only enzyme that metabolizes 5-formylTHF and catalyzes its ATP-dependent conversion to 5,10-methenylTHF. This reaction determines intracellular 5-formylTHF concentrations and converts 5-formylTHF into an enzyme cofactor. The regulation and metabolic role of MTHFS in one-carbon metabolism was investigated in vitro and in human neuroblastoma cells. Steady-state kinetic studies revealed that 10 formylTHF, which exists in chemical equilibrium with 5,10-methenylTHF, acts as a tight binding inhibitor of mouse MTHFS. [6R]-10-formylTHF inhibited MTHFS with a K(i) of 150 nM, and [6R,S]-10-formylTHF triglutamate inhibited MTHFS with a K(i) of 30 nm. MTHFS is the first identified 10-formylTHF tight-binding protein. Isotope tracer studies in neuroblastoma demonstrate that MTHFS enhances de novo purine biosynthesis, indicating that MTHFS-bound 10-formylTHF facilitates de novo purine biosynthesis. Feedback metabolic regulation of MTHFS by 10-formylTHF indicates that 5-formylTHF can only accumulate in the presence of 10-formylTHF, providing the first evidence that 5-formylTHF is a storage form of excess formylated folates in mammalian cells. The sequestration of 10-formylTHF by MTHFS may explain why de novo purine biosynthesis is protected from common disruptions in the folate-dependent one-carbon network. PMID- 16365038 TI - The regulatory domain of SRK2E/OST1/SnRK2.6 interacts with ABI1 and integrates abscisic acid (ABA) and osmotic stress signals controlling stomatal closure in Arabidopsis. AB - ABI1 and ABI2 encode PP2C-type protein phosphatases and are thought to negatively regulate many aspects of abscisic acid (ABA) signaling, including stomatal closure in Arabidopsis. In contrast, SRK2E/OST1/SnRK2.6 encodes an Arabidopsis SnRK2 protein kinase and acts as a positive regulator in the ABA-induced stomatal closure. SRK2E/OST1 is activated by osmotic stress as well as by ABA, but the independence of the two activation processes has not yet been determined. Additionally, interaction between SRK2E/OST1 and PP2C-type phosphatases (ABI1 and ABI2) is not understood. In the present study, we demonstrated that the abi1-1 mutation, but not the abi2-1 mutation, strongly inhibited ABA-dependent SRK2E/OST1 activation. In contrast, osmotic stress activated SRK2E/OST1 even in abi1-1 and aba2-1 plants. The C-terminal regulatory domain of SRK2E/OST1 was required for its activation by both ABA and osmotic stress in Arabidopsis. The C terminal domain was functionally divided into Domains I and II. Domain II was required only for the ABA-dependent activation of SRK2E/OST1, whereas Domain I was responsible for the ABA-independent activation. Full-length SRK2E/OST1 completely complemented the wilty phenotype of the srk2e mutant, but SRK2E/OST1 lacking Domain II did not. Domain II interacted with the ABI1 protein in a yeast two-hybrid assay. Our results suggested that the direct interaction between SRK2E/OST1 and ABI1 through Domain II plays a critical role in the control of stomatal closure. PMID- 16365039 TI - Intracellular inclusions containing mutant alpha1-antitrypsin Z are propagated in the absence of autophagic activity. AB - Mutant alpha(1)-antitrypsin Z (alpha(1)-ATZ) protein, which has a tendency to form aggregated polymers as it accumulates within the endoplasmic reticulum of the liver cells, is associated with the development of chronic liver injury and hepatocellular carcinoma in hereditary alpha(1)-antitrypsin (alpha(1)-AT) deficiency. Previous studies have suggested that efficient intracellular degradation of alpha(1)-ATZ is correlated with protection from liver disease in alpha(1)-AT deficiency and that the ubiquitin-proteasome system accounts for a major route, but not the sole route, of alpha(1)-ATZ disposal. Yet another intracellular degradation system, autophagy, has also been implicated in the pathophysiology of alpha(1)-AT deficiency. To provide genetic evidence for autophagy-mediated disposal of alpha(1)-ATZ, here we used cell lines deleted for the Atg5 gene that is necessary for initiation of autophagy. In the absence of autophagy, the degradation of alpha(1)-ATZ was retarded, and the characteristic cellular inclusions of alpha(1)-ATZ accumulated. In wild-type cells, colocalization of the autophagosomal membrane marker GFP-LC3 and alpha(1)-ATZ was observed, and this colocalization was enhanced when clearance of autophagosomes was prevented by inhibiting fusion between autophagosome and lysosome. By using a transgenic mouse with liver-specific inducible expression of alpha(1)-ATZ mated to the GFP-LC3 mouse, we also found that expression of alpha(1)-ATZ in the liver in vivo is sufficient to induce autophagy. These data provide definitive evidence that autophagy can participate in the quality control/degradative pathway for alpha(1)-ATZ and suggest that autophagic degradation plays a fundamental role in preventing toxic accumulation of alpha(1)-ATZ. PMID- 16365040 TI - Laminin alpha5 is required for dental epithelium growth and polarity and the development of tooth bud and shape. AB - In tooth development, the oral ectoderm and mesenchyme coordinately and reciprocally interact through the basement membrane for their growth and differentiation to form the proper shape and size of the tooth. Laminin alpha5 subunit-containing laminin-10/11 (LM-511/521) is the major laminin in the tooth germ basement membrane. Here, we have examined the role of laminin alpha5 (Lama5) in tooth development using laminin alpha5-null mouse primary dental epithelium and tooth germ organ cultures. Lama5-null mice develop a small tooth germ with defective cusp formation and have reduced proliferation of dental epithelium. Also, cell polarity and formation of the monolayer of the inner dental epithelium are disturbed. The enamel knot, a signaling center for tooth germ development, is defective, and there is a significant reduction of Shh and Fgf4 expression in the dental epithelium. In the absence of laminin alpha5, the basement membrane in the inner dental epithelium becomes discontinuous. In normal mice, integrin alpha6beta4, a receptor for laminin alpha5, is strongly localized at the basal layer of the epithelium, whereas in mutant mice, integrin alpha6beta4 is expressed around the cell surface. In primary dental epithelium culture, laminin 10/11 promotes cell growth, spreading, and filopodia-like microspike formation. This promotion is inhibited by anti-integrin alpha6 and beta4 antibodies and by phosphatidylinositol 3-kinase inhibitors and dominant negative Rho-GTPase family proteins Cdc42 and Rac. In organ culture, anti-integrin alpha6 antibody and wortmannin reduce tooth germ size and shape. Our studies demonstrate that laminin alpha5 is required for the proliferation and polarity of basal epithelial cells and suggest that the interaction between laminin-10/11-integrin alpha6beta4 and the phosphatidylinositol 3-kinase-Cdc42/Rac pathways play an important role in determining the size and shape of tooth germ. PMID- 16365041 TI - Cathepsin S controls angiogenesis and tumor growth via matrix-derived angiogenic factors. AB - The cysteine protease cathepsin S is highly expressed in malignant tissues. By using a mouse model of multistage murine pancreatic islet cell carcinogenesis in which cysteine cathepsin activity has been functionally implicated, we demonstrated that selective cathepsin S deficiency impaired angiogenesis and tumor cell proliferation, thereby impairing angiogenic islet formation and the growth of solid tumors, whereas the absence of its endogenous inhibitor cystatin C resulted in opposite phenotypes. Although mitogenic vascular endothelial growth factor, transforming growth factor-beta1, and the anti-angiogenic endostatin levels in either serum or carcinoma tissue extracts did not change in cathepsin S or cystatin C-null mice, tumor tissue basic fibroblast growth factor and serum type 1 insulin growth factor levels were higher in cystatin C-null mice, and serum type 1 insulin growth factor levels were also increased in cathepsin S-null mice. Furthermore, cathepsin S affected the production of type IV collagen derived anti-angiogenic peptides and the generation of bioactive pro-angiogenic gamma2 fragments from laminin-5, revealing a functional role for cathepsin S in angiogenesis and neoplastic progression. PMID- 16365043 TI - Oligomeric structure of the carnitine transporter CaiT from Escherichia coli. AB - The carnitine transporter CaiT from Escherichia coli belongs to the betaine, choline, and carnitine transporter family of secondary transporters. It acts as an L-carnitine/gamma-butyrobetaine exchanger and is predicted to span the membrane 12 times. Unlike the other members of this transporter family, it does not require an ion gradient and does not respond to osmotic stress (Jung, H., Buchholz, M., Clausen, J., Nietschke, M., Revermann, A., Schmid, R., and Jung, K. (2002) J. Biol. Chem. 277, 39251-39258). The structure and oligomeric state of the protein was examined in detergent and in lipid bilayers. Blue native gel electrophoresis indicated that CaiT was a trimer in detergent solution. This result was further supported by gel filtration and cross-linking studies. Electron microscopy and single particle analysis of the protein showed a triangular structure of three masses or two parallel elongated densities. Reconstitution of CaiT into lipid bilayers yielded two-dimensional crystals that indicated that CaiT was a trimer in the membrane, similar to its homologue BetP. The implications of the trimeric structure on the function of CaiT are discussed. PMID- 16365042 TI - Targeted overexpression of sarcolipin in the mouse heart decreases sarcoplasmic reticulum calcium transport and cardiac contractility. AB - The role of sarcolipin (SLN) in cardiac physiology was critically evaluated by generating a transgenic (TG) mouse model in which the SLN to sarco(endoplasmic)reticulum (SR) Ca(2+) ATPase (SERCA) ratio was increased in the ventricle. Overexpression of SLN decreases SR calcium transport function and results in decreased calcium transient amplitude and rate of relaxation. SLN TG hearts exhibit a significant decrease in rates of contraction and relaxation when assessed by ex vivo work-performing heart preparations. Similar results were also observed with muscle preparations and myocytes from SLN TG ventricles. Interestingly, the inhibitory effect of SLN was partially relieved upon high dose of isoproterenol treatment and stimulation at high frequency. Biochemical analyses show that an increase in SLN level does not affect PLB levels, monomer to pentamer ratio, or its phosphorylation status. No compensatory changes were seen in the expression of other calcium-handling proteins. These studies suggest that the SLN effect on SERCA pump is direct and is not mediated through increased monomerization of PLB or by a change in PLB phosphorylation status. We conclude that SLN is a novel regulator of SERCA pump activity, and its inhibitory effect can be reversed by beta-adrenergic agonists. PMID- 16365044 TI - Follicular fluid high density lipoprotein-associated sphingosine 1-phosphate is a novel mediator of ovarian angiogenesis. AB - Angiogenesis plays an important role in the development of the ovarian follicle and its subsequent transition into the corpus luteum. Accordingly, follicular fluid is a rich source of mitogenic and angiogenic factors such as basic fibroblast growth factor and vascular endothelial growth factor secreted by granulosa cells. In the present study, we show that follicular fluid deprived of basic fibroblast growth factor or vascular endothelial growth factor by means of thermal denaturation or antibody neutralization retains its capacity to stimulate endothelial proliferation and angiogenesis. Mass spectrometric analysis of chromatographic fractions stimulating endothelial growth obtained from follicular fluid revealed that the heat-stable mitogenic activity is identical with the subfraction alpha of high density lipoproteins purified from follicular fluid (FF HDL). Further investigations demonstrated that sphingosine 1-phosphate (S1P), one of the lysophospholipids associated with HDL, accounts for the capacity of this lipoprotein to stimulate endothelial growth and the formation of new vessels. Activation of mitogen-activated protein kinase (p42/44(ERK1/2)), protein kinase C, and protein kinase Akt represent signaling pathways utilized by FF-HDL and S1P to induce endothelial proliferation and angiogenesis. We conclude that FF-HDL represents a novel mitogenic and angiogenic factor present in follicular fluid and that S1P is one of the FF-HDL lipid components accounting for this activity. PMID- 16365045 TI - The low density lipoprotein receptor-related protein 6 interacts with glycogen synthase kinase 3 and attenuates activity. AB - Glycogen synthase kinase 3 (GSK3) is a widely expressed Ser/Thr protein kinase that phosphorylates numerous substrates. This large number of substrates requires precise and specific regulation of GSK3 activity, which is achieved by a combination of phosphorylation, localization, and interactions with GSK3-binding proteins. Members of the Wnt canonical pathway have been shown to influence GSK3 activity. Through a yeast two-hybrid screen, we identified the Wnt canonical pathway co-receptor protein low density lipoprotein receptor-related protein 6 (LRP6) as a GSK3-binding protein. The interaction between the C terminus of LRP6 and GSK3 was also confirmed by in vitro GST pull-down assays and in situ coimmunoprecipitation assays. In vitro assays using immunoprecipitated proteins demonstrated that the C terminus of LRP6 significantly attenuated the activity of GSK3beta. In situ, LRP6 significantly decreased GSK3beta-mediated phosphorylation of tau at both primed and unprimed sites. Finally, it was also demonstrated that GSK3beta phosphorylates the PPP(S/T)P motifs in the C terminus of LRP6. This is the first identification of a direct interaction between LRP6 and GSK3, which results in an attenuation of GSK3 activity. PMID- 16365046 TI - Activation of the checkpoint kinase Rad53 by the phosphatidyl inositol kinase like kinase Mec1. AB - Saccharomyces cerevisiae Rad53, the ortholog of mammalian Chk2, is an essential protein kinase in DNA damage and DNA replication checkpoint pathways. Consecutive phosphatidyl inositol kinase-like kinase (PIKK)-dependent and PIKK-independent steps in activation of Rad53 are key steps for controlling and transmitting diverse downstream responses to DNA damage. However, these activities have not been demonstrated in vitro in defined systems. Here, we have shown that enzymatically dephosphorylated purified Rad53 autoactivates in vitro through a phosphorylation-dependent mechanism. Kinetic analysis demonstrated that autophosphorylation results in a more than 9-fold increase in protein kinase activity. Autophosphorylation was Rad53 concentration-dependent, indicating that the reaction follows an intermolecular mechanism. DNA damage induced oligomerization of a subset of Rad53 molecules in vivo. At low concentrations of Rad53, preincubation of Rad53 with immune complexes containing the Mec1/Ddc2 complex can activate Rad53 kinase activity. Our findings showed that Mec1/Ddc2 complexes can directly activate Rad53 through a phosphorylation-dependent mechanism, and more generally, supported the hypothesis that PIKKs regulate Chk2 orthologs through phosphorylation. Moreover, this work has substantiated a model for PIKK-independent amplification of Rad53 activation (and by extension, activation of other Chk2 orthologs) mediated by inter-Rad53 phosphorylation. PMID- 16365047 TI - Prolyl-isomerase Pin1 accumulates in lewy bodies of parkinson disease and facilitates formation of alpha-synuclein inclusions. AB - Parkinson disease (PD) is a relatively common neurodegenerative disorder that is characterized by the loss of dopaminergic neurons and by the formation of Lewy bodies (LBs), which are cytoplasmic inclusions containing aggregates of alpha synuclein. Although certain post-translational modifications of alpha-synuclein and its related proteins are implicated in the genesis of LBs, the specific molecular mechanisms that both regulate these processes and initiate subsequent inclusion body formation are not yet well understood. We demonstrate in our current study, however, that the prolyl-isomerase Pin1 localizes to the LBs in PD brain tissue and thereby enhances the formation of alpha-synuclein immunoreactive inclusions. Immunohistochemical analysis of brain tissue from PD patients revealed that Pin1 localizes to 50-60% of the LBs that show an intense halo pattern resembling that of alpha-synuclein. By utilizing a cellular model of alpha-synuclein aggregation, we also demonstrate that, whereas Pin1 overexpression facilitates the formation of alpha-synuclein inclusions, dominant negative Pin1 expression significantly suppresses this process. Consistent with these observations, Pin1 overexpression enhances the protein half-life and insolubility of alpha-synuclein. Finally, we show that Pin1 binds synphilin-1, an alpha-synuclein partner, via its Ser-211-Pro and Ser-215-Pro motifs, and enhances its interaction with alpha-synuclein, thus likely facilitating the formation of alpha-synuclein inclusions. These results indicate that Pin1-mediated prolyl isomerization plays a pivotal role in a post-translational modification pathway for alpha-synuclein aggregation and in the resultant Lewy body formations in PD. PMID- 16365048 TI - Target selectivity of vertebrate notch proteins. Collaboration between discrete domains and CSL-binding site architecture determines activation probability. AB - All four mammalian Notch proteins interact with a single DNA-binding protein (RBP jkappa), yet they are not equivalent in activating target genes. Parallel assays of three Notch-responsive promoters in several cell lines revealed that relative activation strength is dependent on protein module and promoter context more than the cellular context. Each Notch protein reads binding site orientation and distribution on the promoter differently; Notch1 performs extremely well on paired sites, and Notch3 prefers single sites in conjunction with a proximal zinc finger transcription factor. Although head-head sites can elicit a Notch response on their own, use of CBS (CSL binding site) in tail-tail orientation is context dependent. Bias for specific DNA elements is achieved by interplay between the N terminal RAM (RBP-jkappa-associated molecule/ankyrin region), which interprets CBS proximity and orientation, and the C-terminal transactivation domain that interacts specifically with the transcription machinery or nearby factors. To confirm the prediction that modular design underscores the evolution of functional divergence between Notch proteins, we generated a synthetic Notch protein (Notch1 ankyrin with Notch3 transactivation domain) that displayed superior signaling strength on the hes5 promoter. Consistent with the prediction that "preferred" targets (Hes1) should respond faster and at lower Notch concentration than other targets, we showed that Hes5-GFP was extinguished fast and recovered slowly, whereas Hes1-GFP was inhibited late and recovered quickly after a pulse of DAPT in metanephroi cultures. PMID- 16365049 TI - Nutritional studies in Victorian prisons. PMID- 16365050 TI - Helen Gertrude Oldham (1903-2004). PMID- 16365051 TI - Induction of hepatic antioxidant enzymes by phenolic acids in rats is accompanied by increased levels of multidrug resistance-associated protein 3 mRNA expression. AB - Phenolic acids are widespread in plant foods; they contain important biological and pharmacological properties, some of which were shown to be effective in preventing cancer. We investigated the modulatory effects of phenolic acids on an antioxidant system in male Sprague-Dawley rats. Rats were orally administrated gentisic acid (GEA), gallic acid (GA), ferulic acid (FA), and p-coumaric acid (p CA) at a dosage of 100 mg/kg body weight for 14 consecutive days. At this dose, the activities of hepatic superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase were greater after administration of all 4 phenolic acids compared with the control group (P < 0.05). The activities of these enzymes in the small intestine of rats were also significantly greater after GA and p-CA treatment compared with controls. The changes in hepatic CuZnSOD, GPx, and catalase mRNA levels induced by phenolic acids were similar to those noted in the enzyme activities. Oxidized glutathione levels were lower (P < 0.05) in the liver of all phenolic acid-supplemented rats, whereas reduced glutathione was markedly higher than in control rats, especially after administration of GA and p-CA. The liver homogenates obtained from rats that had been administered phenolic acids had higher oxygen radical absorbance capacity than those obtained from control rats. Immunoblot analysis revealed an increased total level of Nrf2, a transcription factor governing the antioxidant response element in phenolic acid supplemented rats. Phenolic acid-mediated antioxidant enzyme expression was accompanied by upregulation of multidrug resistance-associated protein Mrp3. These experiments show that modulation of phase II antioxidant enzymes and oxidative status in the liver by phenolic acids may play an important role in the protection against adverse effects related to mutagenesis and oxidative damage. PMID- 16365052 TI - Identification of the regulatory region of the L-type pyruvate kinase gene in mouse liver by hydrodynamics-based gene transfection. AB - Expression of L-type pyruvate kinase (L-PK) is upregulated in the liver by dietary carbohydrate. Previously, 3 carbohydrate/insulin response elements were identified in the 5'-flanking region of the L-PK gene up to bp -170. Studies of the 5'-flanking region beyond bp -183 in transgenic mice suggested that other regulatory elements may be present upstream of bp -183, but the positions of these elements were uncertain. In the present study, the existence of regulatory regions of the L-PK gene responding to stimulation by feeding was examined using in vivo hydrodynamics-based gene transfection (HT) in mouse liver. The firefly luciferase (FL) gene, fused with various lengths of the 5'-flanking region of the L-PK gene, was introduced into mouse liver by HT. The mice had free access to a high-carbohydrate diet. In liver homogenate, luciferase activity of pL-PK(-1467) FL (which included the 5'-flanking region from bp -1467 to +17), was markedly stimulated by feeding. 5'-Deletion up to bp -1065 caused only minor changes in luciferase activity, but further deletion up to bp -690 and bp -203 caused significant, gradual decreases in activity. Further analyses utilizing 5' deletion mutants indicated the existence of positive regulatory regions that respond to stimulation by feeding between bp -1065 and -945, and between -300 and -203 on the L-PK gene. These results suggest that unidentified cis-acting DNA elements exist in the upstream region of the L-PK gene, and that HT is a useful approach for detecting regulatory regions of genes expressed in the liver. PMID- 16365053 TI - Copper transport protein (Ctr1) levels in mice are tissue specific and dependent on copper status. AB - Studies were conducted to determine distribution of the copper transporter, Ctr1, a transmembrane protein responsible for cellular copper uptake, in adult mice and in suckling mice nursed by either copper-adequate (Cu+) or copper-deficient (Cu-) dams. Western immunoblot analyses, using immunopurified antibody, detected monomeric (23 kDa) and oligomeric forms of Ctr1 in the membrane fraction of several mouse organs. Immunohistochemical analyses detected abundant Ctr1 protein in liver canaliculi; kidney cortex tubules; small intestinal enterocytes; the choroid plexus and capillaries of brain; intercalated disks of heart; mature spermatozoa; epithelium of mammary ducts; and the pigment epithelium, outer limiting membrane, and outer plexiform layer of the retina. Duodenal Ctr1 distribution was different in the adult compared with suckling mice; adult mice demonstrated strong intracellular staining of the enterocyte, whereas apical staining predominated in suckling mice. In Cu- mice at postnatal d 16 (P16), Ctr1 staining was augmented in kidney, duodenum, and choroid plexus, compared with Cu+ mice. Brain immunoblot data indicated that Ctr1 protein in membrane fractions of Cu- mice was 56% higher than Cu+ mice. Cu- mice had lower hemoglobin (56% of Cu+), and lower copper concentration (% of Cu+) in liver (15%), brain (26%), and kidney (65%). These results suggest that Ctr1 protein is expressed in multiple tissues and found in higher levels in selected organs after perinatal copper deficiency. Enhanced Ctr1 levels and redistribution might compensate in part for the decrease in copper supply. Mechanisms for the enhancement in Ctr1 staining remain to be established. PMID- 16365054 TI - Rapamycin inhibits liver growth during refeeding in rats via control of ribosomal protein translation but not cap-dependent translation initiation. AB - We examined the role of the mammalian target of rapamycin (mTOR) in hepatic cell growth. To dissociate cell growth from cell proliferation, we employed an in vivo model of nonproliferative liver growth in rats, refeeding after 48 h of food deprivation. Starvation resulted in a decrease in liver mass, liver protein, and cell size, all of which were largely restored after 24 h of refeeding. Administration of the mTOR inhibitor, rapamycin, before the refeeding period partially inhibited the restoration of liver protein content. Refeeding was also associated with an increase in ribosomal protein S6 phosphorylation and phosphorylation of the eukaryotic initiation factor (eIF) 4E binding protein 1 (4E-BP1). 4E-BP1 phosphorylation was accompanied by a decrease in the abundance of the complex containing 4E-BP1 with eIF4E. These changes were prevented by rapamycin administration. However, association of eIF4E and eIF4G and eIF2alpha phosphorylation, both of which are stimulated by refeeding, were insensitive to rapamycin. The functional importance of these observations was confirmed by polysome fractionation, which showed that translation initiation of 5' oligopyrimidine tract-containing mRNAs, which encode ribosomal proteins, was inhibited by rapamycin, whereas translation of signal transducer and activator of transcription 1 (STAT1), a cap-dependent mRNA, was unaffected. The abundance of ribosomal proteins paralleled total protein content during refeeding in both control and rapamycin-injected rats. We conclude that accretion of liver protein during refeeding is dependent on mTOR-mediated activation of the translation of ribosomal proteins but not dependent on mTOR-mediated activation of cap-dependent translation initiation. PMID- 16365055 TI - Orally administered betaine has an acute and dose-dependent effect on serum betaine and plasma homocysteine concentrations in healthy humans. AB - Betaine, i.e., trimethylglycine, is linked to homocysteine metabolism. A 3-mo daily betaine supplementation decreased even normal plasma total homocysteine (tHcy) concentrations in humans. The pharmacokinetic characteristics and metabolism of betaine in humans have not been investigated in detail. The aim of this study was to assess the pharmacokinetics of orally administered betaine and its acute effect on plasma tHcy concentrations. Healthy volunteers (n = 10; 3 men, 7 women) with normal body weight (mean +/- SD, 69.5 +/- 17.0 kg), 40.8 +/- 12.4 y old, participated in the study. The betaine doses were 1, 3, and 6 g. The doses were mixed with 150 mL of orange juice and ingested after a 12-h overnight fast by each volunteer according to a randomized double-blind crossover design. Blood samples were drawn for 24 h and a 24-h urine collection was performed. Orally administered betaine had an immediate and dose-dependent effect on serum betaine concentration. Single doses of 3 and 6 g lowered plasma tHcy concentrations (P = 0.019 and P < 0.001, respectively), unlike the 1-g dose. After the highest dose, the concentrations remained low during the 24 h of monitoring. The change in plasma tHcy concentration was linearly associated with betaine dose (P = 0.006) and serum betaine concentration (R2 = 0.17, P = 0.025). The absorption and elimination of betaine were dose dependent. The urinary excretion of betaine seemed to increase with an increasing betaine dose, although a very small proportion of ingested betaine was excreted via urine. In conclusion, a single dose of orally administered betaine had an acute and dose dependent effect on serum betaine concentration and resulted in lowered plasma tHcy concentrations within 2 h in healthy subjects. PMID- 16365056 TI - Rat gastrointestinal tissues metabolize quercetin. AB - Quercetin and quercetin glycosides from food or dietary supplements appear in body tissues almost exclusively as glucuronated, sulfated, and methylated quercetin conjugates, suggesting that the in vivo bioactivity of quercetin may be due to its metabolites. In this study, pre- and postabsorptive metabolism of orally ingested quercetin was examined by comparing the metabolite pattern in gastrointestinal (GI) tissues, contents, and internal tissues. F344 rats (n = 6) were fed for 6 wk a diet containing 0.45% quercetin and the metabolite patterns were determined in the tissues and contents of stomach, small intestine, cecum, and colon and in liver, kidney, and plasma using LC-MS/MS. GI contents contained predominantly unmetabolized quercetin at 94-100%, whereas quercetin in GI tissues was present as 11 different sulfated, glucuronated, and methylated metabolites at 32% in stomach, 88% in small intestine, 27% in cecum, and 46% in colon. Quercetin was further metabolized postabsorption and found in liver, kidney, and plasma almost exclusively as sulfated methyl-quercetin glucuronide. The unique pattern of quercetin metabolites in each GI tissue indicates extensive biotransformation before absorption and distribution in rats. PMID- 16365057 TI - Factors affecting the bioavailability of soy isoflavones in humans after ingestion of physiologically relevant levels from different soy foods. AB - The precise role that isoflavones play in the health-related effects of soy foods, and their potential for adverse effects are controversial. This may be due in part to a lack of basic knowledge regarding their bioavailability and metabolism, particularly as it relates to the soy source. To date, there is little information concerning possible differences in the bioavailability of isoflavones derived from natural soy foods consumed at physiologically relevant intakes and whether age- or gender-related differences influence that bioavailability. In the current study of healthy adults [premenopausal (n = 21) and postmenopausal (n = 17) women and a group of men (n = 21)], we examined the effect of age, gender, and the food matrix on the bioavailability of isoflavones for both the aglycon and glucoside forms that are naturally present in 3 different soy foods, soy milk, textured vegetable protein, and tempeh. The study was designed as a random crossover trial so that all individuals received each of the 3 foods. The dose of isoflavones administered to each individual as a single bolus dose was 0.44 mg/kg body weight. Pharmacokinetic parameters were normalized to mg of each isoflavone ingested per kilogram body weight to account for differences in daidzein and genistein content between the diets. Serum isoflavone concentrations in all individuals and groups increased rapidly after the ingestion of each soy food; as expected, genistein concentrations exceeded daidzein concentrations in serum. In this small study, gender differences in peak concentrations of daidzein were observed, with higher levels attained in women. Consumption of tempeh (mainly isoflavone aglycon) resulted in higher serum peak levels of both daidzein (P < 0.001) and genistein (P < 0.01) and the associated area under the curve (P < 0.001 and P < 0.03, respectively) compared with textured vegetable protein (predominantly isoflavone glucosides). However, soy milk was absorbed faster and peak levels of isoflavones were attained earlier than with the other soy foods. Only 30% of the subjects were equol producers and no differences in equol production with age or gender were observed. PMID- 16365058 TI - Of the major phenolic acids formed during human microbial fermentation of tea, citrus, and soy flavonoid supplements, only 3,4-dihydroxyphenylacetic acid has antiproliferative activity. AB - Dietary flavonoids are poorly absorbed from the gastrointestinal tract. Colonic bacteria convert flavonoids into smaller phenolic acids (PA), which can be absorbed into the circulation and may contribute to the chemopreventive activity of the parent compounds. The purpose of our study was to determine whether flavonoids from green and black tea (GT, BT), citrus fruit with rutin (CF+R) and soy (S) supplements exposed to the same conditions in a dynamic in vitro model of the colon (TIM-2) will form the same phenolic acid products of microbial metabolism. About 600 mg of flavonoids from GT, BT, CF+R and S extracts were infused at t = 0 and 12 h into the TIM-2. Samples from the lumen and dialysate were collected at t = 0,4,8,12,16,24 and 28h. The flavonoid and PA concentrations were measured by HPLC and GC-MS. GT, BT, and CF+R formed 3-methoxy-4 hydroxyphenylacetic acid (3M4HPAA), 4-hydroxyphenyl acetic acid (4HPAA), 3,4 dihydroxyphenylacetic acid (3,4DHPAA), and 3-(3-hydroxyphenyl) propionic acid (3,3HPPA). BT flavonoids were also metabolized to 2,4,6-trihydroxybenzoic acid (2,4,6THBA) and CF+R flavonoids to 3-(4-hydroxy-3-methoxyphenyl) propionic acid (3,4H3MPPA), 3-hydroxyphenyl acetic acid (3HPAA) and a small amount of hippuric acid. After S infusion, we found 3M4HPAA and 4HPAA only. Among these phenolic acids, only 3,4DHPAA exhibited antiproliferative activity in prostate and colon cancer cells. 3,4DHPAA was significantly (P < 0.005) more inhibitory in colon cancer cells (HCT116) compared with an immortalized normal intestinal epithelial cell line (IEC6). In summary, fermentation by intestinal microbes of GT, BT, C+R, and S flavonoids resulted in the conversion to the same major phenolic acids. PMID- 16365059 TI - Colonic fermentation may play a role in lactose intolerance in humans. AB - The results of our previous study suggested that in addition to the small intestinal lactase activity and transit time, colonic processing of lactose may play a role in lactose intolerance. We investigated whether colonic fermentation of lactose is correlated with lactose intolerance. After 28 Chinese subjects had undergone 1 glucose (placebo) and 2 lactose challenges, consistent lactose tolerant (n = 7) and intolerant (n = 5) subjects with no complaints after glucose administration were classified on the basis of the 6-h symptom scores. Before the challenges, fecal samples were collected for in vitro incubation with lactose. The incubation was carried out in a static system under anaerobic conditions for 5 h during which samples were taken for measurement of short-chain fatty acids, lactate, lactose, glucose, and galactose. Fecal bacterial composition was determined by fluorescent in situ hybridization. The tolerant and intolerant groups did not differ in the rate or degree of hydrolysis of lactose or production of glucose and galactose. The intolerant group produced d- and l lactate, acetate, propionate, and butyrate significantly faster than the tolerant group. In the intolerant group, the amounts of acetate, propionate, butyrate, and l-lactate produced were higher than those in the tolerant group. Fecal bacterial composition did not differ between the 2 groups. The results indicate that the degree and rate of lactose hydrolysis in the colon do not play a role in lactose intolerance. However, after lactose is hydrolyzed, a faster and higher production of microbial intermediate and end metabolites may be related to the occurrence of symptoms. PMID- 16365060 TI - Increased colonic luminal synthesis of butyric acid is associated with lowered colonic cell proliferation in piglets. AB - Butyrate inhibits colonic cell proliferation in vitro but reportedly has an opposite effect in vivo. Because lactulose feeding decreases cecal cell proliferation, an effect attenuated by prefeeding inulin, we hypothesized that lactulose feeding would decrease colonic luminal synthesis of butyrate, and that prefeeding and cofeeding inulin would prevent this effect. Piglets (n = 31) were catheterized and randomly assigned to 1 of 4 groups: Control formula (C); control formula + lactulose (L); control formula + lactulose + inulin (L + I); and control formula + inulin (I). At 6 and 7 d postsurgery, the rate of cecal synthesis of butyrate, cecal cell proliferation and apoptosis, and cecal and distal colon butyrate concentration were measured. In groups C, L, L + I, and I, the rates of synthesis of butyrate (mean +/- SEM) were 10.6 +/- 3.2, 23.3 +/- 4.5, 12.4 +/- 3.6, and 14.6 +/- 4.0 micromol/min, respectively (Group Effect, P = 0.1; C vs. L, P = 0.03; L vs. L + I, P = 0.06). The cecal butyrate concentrations did not differ among the 4 groups and were 8.7 +/- 3.2, 2.4 +/- 0.8, 3.4 +/- 1.9, and 2.0 +/- 0.7 micromol/g dry wt, respectively. The total cecal cell proliferation index was higher in C than in L (P = 0.008) or I (P = 0.026) and was higher in L + I than in L (P = 0.013) or I (P = 0.046). The increased supply of butyrate to the cecum was associated with decreased cell proliferation, but cecal butyrate concentration did not reflect synthesis. PMID- 16365061 TI - Dietary fructooligosaccharides affect intestinal barrier function in healthy men. AB - In contrast to most expectations, we showed previously that dietary fructooligosaccharides (FOS) stimulate intestinal colonization and translocation of invasive Salmonella enteritidis in rats. Even before infection, FOS increased the cytotoxicity of fecal water, mucin excretion, and intestinal permeability. In the present study, we tested whether FOS has these effects in humans. A double blind, placebo-controlled, crossover study of 2 x 2 wk, with a washout period of 2 wk, was performed with 34 healthy men. Each day, subjects consumed lemonade containing either 20 g FOS or placebo and the intestinal permeability marker chromium EDTA (CrEDTA). On the last 2 d of each supplement period, subjects scored their gastrointestinal complaints on a visual analog scale and collected feces and urine for 24 h. Fecal lactic acid was measured using a colorimetric enzymatic kit. The cytotoxicity of fecal water was determined with an in vitro bioassay, fecal mucins were quantified fluorimetrically, and intestinal permeability was determined by measuring urinary CrEDTA excretion. In agreement with our animal studies, FOS fermentation increased fecal wet weight, bifidobacteria, lactobacilli, and lactic acid. Consumption of FOS increased flatulence and intestinal bloating. In addition, FOS consumption doubled fecal mucin excretion, indicating mucosal irritation. However, FOS did not affect the cytotoxicity of fecal water and intestinal permeability. The FOS-induced increase in mucin excretion in our human study suggests mucosal irritation in humans, but the overall effects are more moderate than those in rats. PMID- 16365062 TI - Soy isoflavones alter expression of genes associated with cancer progression, including interleukin-8, in androgen-independent PC-3 human prostate cancer cells. AB - High consumption of soy isoflavones in Asian diets has been correlated with a lower incidence of clinically important cases of prostate cancer. The chemopreventive properties of these diets may result from an interaction of several types of isoflavones, including genistein and daidzein. The present study investigated the effects of a soy isoflavone concentrate (ISF) on growth and gene expression profiles of PC-3 human prostate cancer cells. Trypan blue exclusion and [3H]-thymidine incorporation assays showed that ISF decreased cell viability and caused a dose-dependent inhibition of DNA synthesis, respectively, with 50% inhibition (IC50) of DNA synthesis at 52 mg/L (P = 0.05). The glucoside conjugates of genistein and daidzein in ISF were converted to bioactive free aglycones in cell culture in association with the inhibition of DNA synthesis. Flow cytometry and Western immunoblot analyses showed that ISF at 200 mg/L caused an accumulation of cells in the G2/M phase of the cell cycle (P < 0.05) and decreased cyclin A by 20% (P < 0.05), respectively. The effect of ISF on the gene expression profile of PC-3 cells was analyzed using Affymetrix oligonucleotide DNA microarrays that interrogate approximately 17,000 human genes. Of the 75 genes altered by ISF, 28 were upregulated and 47 were downregulated (P < 0.05). Further analysis showed that IL-8, matrix metalloproteinase 13, inhibin beta A, follistatin, and fibronectin mRNA levels were significantly reduced, whereas the expression of p21(CIP1), a major cell cycle inhibitory protein, was increased. The effects of ISF on the expression of IL-8 and p21(CIP1) mRNA and protein were validated at high and low ISF concentrations. Our data show that ISF inhibits the growth of PC-3 cells through modulation of cell cycle progression and the expression of genes involved in cell cycle regulation, metastasis, and angiogenesis. PMID- 16365063 TI - Flaxseed oil increases the plasma concentrations of cardioprotective (n-3) fatty acids in humans. AB - Alpha-linolenic acid (ALA) is a major dietary (n-3) fatty acid. ALA is converted to longer-chain (n-3) PUFA, such as eicosapentaenoic acid (EPA) and possibly docosahexaenoic acid (DHA). EPA and DHA are fish-based (n-3) fatty acids that have proven cardioprotective properties. We studied the effect of daily supplementation with 3 g of ALA on the plasma concentration of long-chain (n-3) fatty acids in a predominantly African-American population with chronic illness. In a randomized, double-blind trial, 56 participants were given 3 g ALA/d from flaxseed oil capsules (n = 31) or olive oil placebo capsules (n = 25). Plasma EPA levels at 12 wk in the flaxseed oil group increased by 60%, from 24.09 +/- 16.71 to 38.56 +/- 28.92 micromol/L (P = 0.004), whereas no change occurred in the olive oil group. Plasma docosapentaenoic acid (DPA) levels in the flaxseed oil group increased by 25% from 19.94 +/- 9.22 to 27.03 +/- 17.17 micromol/L (P = 0.03) with no change in the olive oil group. Plasma DHA levels did not change in either group. This study demonstrates the efficacy of the conversion of ALA to EPA and DPA in a minority population with chronic disease. ALA may be an alternative to fish oil; however, additional clinical trials with ALA are warranted. PMID- 16365064 TI - Beef tallow increases the potency of conjugated linoleic acid in the reduction of mouse mammary tumor metastasis. AB - Animal studies consistently show that dietary conjugated linoleic acid (CLA) reduces mammary tumorigenesis including metastasis. Relatively low concentrations of CLA are required for those effects, and a threshold level exists above which there is no added reduction. We reasoned that the concentration of CLA required to effectively alter mammary tumor metastasis may be dependent on the type of dietary fat because select fatty acids can enhance or suppress normal or malignant cell growth and metastasis. For this study, the diets (a total of 12 different groups) differed in fatty acid composition but not in energy from fat (40%). In experiments involving spontaneous metastasis, mice were fed for 11 wk; in experiments in which mice were injected i.v. with tumor cells, they were fed for 7 wk. Mice were then assessed for the effect of CLA concentration on mammary tumorigenesis. Mammary tumor growth was not altered, but metastasis was significantly decreased when beef tallow (BT) replaced half of a defined vegetable fat blend (VFB). That blend reflects the typical fat content of a Western diet. In addition, that same VFB:BT diet lowered the concentration of CLA required to significantly decrease mammary tumor metastasis from 0.1% of the diet to 0.05%. A diet in which corn oil replaced half of the VFB did not lower the threshold from 0.1 to 0.05%. In vitro, the main fatty acid in vegetable oil, linoleic acid, reduced the efficacy of CLA toxicity on mammary tumor cells in culture. Alternatively, fatty acids normally found in BT, such as oleic, stearic, and palmitic acids, either did not change or enhanced the cytolytic effects of CLA isomers on mouse mammary tumor cells in culture. These data provide evidence that dietary BT, itself with negligible levels of CLA, may increase the efficacy of dietary CLA in reducing mammary tumorigenesis. PMID- 16365065 TI - Fish oil affects blood pressure and the plasma lipid profile in healthy Danish infants. AB - Animal and epidemiologic studies indicate that early nutrition has lasting effects on metabolism and cardiovascular disease risk. In adults, (n-3) long chain PUFA (LCPUFA) from fish oils improve blood pressure, the lipid profile, and possibly cardiovascular disease mortality. This randomized trial is the first to investigate the effects of fish oil on blood pressure and the lipid profile in infancy. Healthy term 9-mo old infants (n = 83) were randomly assigned to 5 mL fish oil daily or no fish oil for 3 mo and to 2 different milk types. Before and after the intervention, blood pressure was measured with an oscillometric device, and blood was sampled for analysis of erythrocyte fatty acid composition and the plasma lipid profile. This paper examines the effects of the fish oil supplement, with adjustment for the effects of the milk intervention when relevant. The fish oil intervention increased erythrocyte (n-3) LCPUFA content (P < 0.001). At 12 mo, infants administered fish oil had a lower systolic blood pressure [adjusted mean difference (95% CI)] 6.3 mm Hg (0.9, 11.7) (P = 0.02), a 0.51 mmol/L (0.07, 0.95) higher plasma total cholesterol (P = 0.02), and a 0.52 mmol/L (0.02,1.01) higher LDL cholesterol (P = 0.04) than infants not administered fish oil. Plasma triacylglycerol was inversely associated with the erythrocyte content of eicosapentaenoic acid (r = 0.34, P < 0.01), a biomarker of fish oil dose. The observed effects of fish oil are in accordance with findings in adults. The long term health implications warrant further investigation. PMID- 16365066 TI - Consumption of soy isoflavones does not affect plasma total homocysteine or asymmetric dimethylarginine concentrations in healthy postmenopausal women. AB - Postmenopausal women are at increased risk for cardiovascular disease because many risk factors are aggravated by menopause. Phytoestrogens may modulate risk factors favorably, involving mechanisms similar to estrogen. The effect of phytoestrogens on the atherogenic amino acids homocysteine and asymmetric dimethylarginine (ADMA) was investigated in a controlled intervention study in healthy postmenopausal women. A multicenter, double-blind, crossover intervention trial in 89 postmenopausal women from Denmark, Germany, and the UK was performed. Subjects consumed fruit cereal bars with or without soy isoflavones (50 mg/d) for 8 wk each with an 8-wk washout period in between. Urinary phytoestrogens increased significantly after isoflavone intervention (P < 0.001). Isoflavone supplementation did not affect plasma total homocysteine or ADMA. For homocysteine, changes from baseline were 0.32 micromol/L (range: -0.31-0.92; 95% CI 0.13-0.72), and 0.29 micromol/L (range: -0.45-1.09; 95% CI 0.01-0.63, P = 0.286) for isoflavone treatment and placebo, respectively. For ADMA concentrations, changes from baseline were -0.02 micromol/L (range: -0.08-0.03; 95% CI -0.04-0.01, and 0.00 micromol/L (range: -0.05-0.03; 95% CI -0.03-0.01, P = 0.397) for isoflavone treatment and placebo, respectively. There was no association between plasma total homocysteine and ADMA. Changes from baseline in plasma ADMA and folate were negatively correlated (r = -0.18, P = 0.017). These results challenge the overall health effect of isoflavone supplementation in healthy postmenopausal women. PMID- 16365067 TI - Lycopene supplementation prevents smoke-induced changes in p53, p53 phosphorylation, cell proliferation, and apoptosis in the gastric mucosa of ferrets. AB - Cigarette smoking increases the risk for gastric cancer. Higher intakes or blood levels of lycopene are associated with a decreased risk of gastric cancer. However, the biological mechanisms by which lycopene may protect against gastric carcinogenesis are poorly understood. We evaluated the effects of lycopene supplementation on smoke-induced changes in protein levels of p53, p53 target genes (p21(Waf1/Cip1) and Bax-1), cell proliferation, and apoptosis in the gastric mucosa of ferrets. Ferrets were assigned to cigarette smoke exposure or to no exposure and to no, low-dose, or high-dose lycopene supplementation (2 x 3 factorial design) for 9 wk. Lycopene concentrations were significantly elevated in a dose-dependent manner in the gastric mucosa of ferrets supplemented with lycopene alone, but were markedly reduced in ferrets supplemented with lycopene and exposed to smoke. Although ferrets were given lycopene containing 95% all trans isomers, cis isomers were the predominant forms in the gastric mucosa. Total p53 and phosphorylated p53 levels were greater in ferrets exposed to smoke alone than in all other groups. Levels were approximately 300 and 500% of the controls, respectively. However, smoke-elevated total p53 and phosphorylated p53 were markedly attenuated by both doses of lycopene. p21(Waf1/Cip1), Bax-1, and cleaved caspase 3 were substantially decreased, whereas cyclin D1 and proliferating cellular nuclear antigen (PCNA) were increased in ferrets exposed to smoke alone. Lycopene prevented smoke-induced changes in p21(Waf1/Cip1), Bax 1, cleaved caspase 3, cyclin D1, and PCNA in a dose-dependent fashion. These data indicate that lycopene may prevent smoke exposure-induced changes in p53, p53 phosphorylation, p53 target genes, cell proliferation, and apoptosis in the gastric mucosa of ferrets. PMID- 16365068 TI - A lignan complex isolated from flaxseed does not affect plasma lipid concentrations or antioxidant capacity in healthy postmenopausal women. AB - A lignan complex rich in the plant lignan secoisolariciresinol diglucoside (SDG) was isolated from flaxseed. SDG is metabolized by the colonic microflora to the mammalian lignans enterodiol (END) and enterolactone (ENL), and was hypothesized to reduce plasma lipid concentrations and improve antioxidant capacity. The aim of this study was to investigate the effects of a lignan complex, providing 500 mg/d of SDG, on serum concentration and urinary excretion of ENL, plasma lipids, serum lipoprotein oxidation resistance, and markers of antioxidant capacity. Healthy postmenopausal women (n = 22) completed a randomized, double-blind, placebo-controlled, crossover study. Women consumed daily a low-fat muffin, with or without a lignan complex, for 6 wk, separated by a 6-wk washout period. Serum ENL concentration, urinary ENL excretion, plasma concentrations of total cholesterol (TC), LDL cholesterol (LDL-C), HDL cholesterol (HDL-C), triacylglycerol (TAG), serum lipoprotein oxidation lag time, plasma Trolox equivalent antioxidant capacity (TEAC), and ferric reducing ability of plasma (FRAP) were measured at the beginning and end of each intervention period. ENL concentrations in serum (P < 0.001) and ENL urinary excretion (P < 0.001) were significantly higher after the lignan complex intervention period compared with placebo. Plasma concentrations of TC, LDL-C, HDL-C, TAG, lipoprotein oxidation lag time, TEAC and FRAP were not affected. Daily consumption for 6 wk of a low fat muffin enriched with a lignan complex significantly increased serum ENL concentrations and urinary ENL excretion in healthy postmenopausal women, but had no effect on plasma lipid concentrations, serum lipoprotein oxidation resistance, or plasma antioxidant capacity. PMID- 16365069 TI - Dietary inulin intake and age can affect intestinal absorption of zinc and copper in rats. AB - Nondigestible inulin-type fructan intake can stimulate intestinal mineral absorption in both humans and animals. However, this stimulatory effect may depend on experimental conditions such as the duration of the experience, mineral levels in the diet, and the animal's physiological status. The aim of this study was to determine the effect of inulin intake on Zn and Cu absorption in rats at different ages. Male Wistar rats (n = 80) of 4 different ages (2,5, 10, and 20 mo) were randomly assigned to a control group or a group administered 3.75% inulin in their diet for 4 d followed by 7.5% inulin for 26 d. Absorption of Zn67 and Cu65 was determined on d 21 of the experiment by fecal monitoring using Zn67 and Cu65 isotopes. Zn and Cu status was also assessed. Absorption of Zn67 and Cu65 was significantly lower in 11- and 21-mo-old rats than in 3- and 6 mo-old rats. Moreover, inulin intake significantly increased Zn67 and Cu65 absorption. In conclusion, age and dietary inulin intake can significantly affect intestinal absorption of zinc and copper in rats. Further studies are required to explore this effect over longer periods of inulin intake and to test the effects of inulin in humans. PMID- 16365070 TI - Bread fortified with cholecalciferol increases the serum 25-hydroxyvitamin D concentration in women as effectively as a cholecalciferol supplement. AB - Fortification of foods is a feasible way of preventing low vitamin D status. Bread could be a suitable vehicle for fortification because it is a common part of diets worldwide. The bioavailability of cholecalciferol from bread is not known. We studied cholecalciferol stability, the concentration of the added cholecalciferol, the dispersion of cholecalciferol in bread, and the bioavailability of cholecalciferol from fortified bread. Three batches of fortified low-fiber wheat and high-fiber rye breads were baked; from each batch, 3 samples of dough and bread were analyzed for their cholecalciferol content. In a single-blind bioavailability study, 41 healthy women, 25-45 y old, with mean serum 25-hydroxyvitamin D concentration 29 nmol/L (range 12-45 nmol/L), were randomly assigned to 4 study groups. Each group consumed fortified wheat bread, fortified rye bread, regular wheat bread (control), or regular wheat bread and a cholecalciferol supplement (vitamin D control) daily for 3 wk. The daily dose of vitamin D was 10 mug in all groups except the control group. The vitamin dispersed evenly in the breads and was stable. Both fortified breads increased serum 25-hydroxyvitamin D concentration as effectively as the cholecalciferol supplement. Supplementation or fortification did not affect serum intact parathyroid hormone concentration or urinary calcium excretion. In conclusion, fortified bread is a safe and feasible way to improve vitamin D nutrition. PMID- 16365071 TI - Dietary vitamin A has both chronic and acute effects on vitamin A indices in lactating rats and their offspring. AB - To further investigate the effect of dietary vitamin A (VA) intake on milk VA concentrations and pup VA status, female rats were fed 2 concentrations of VA [0 (n = 9) or 50 micromol/kg diet (n = 10)] during pregnancy and lactation. Plasma retinol concentrations were significantly higher (30-40%) during lactation than before pregnancy or after weaning but were not influenced by dietary VA. In rats fed VA, VA concentrations during lactation were significantly higher in milk (1.5 3 times), mammary tissue (>100%), liver (4 times), pup plasma (20-40%), and pup liver (1.1-6.7 times). In Expt. 2, when VA intake was switched on d 7 of lactation from 0 to 50 micromol/kg, milk VA concentrations (2.24 +/- 0.42 micromol/L; mean +/- SD, n = 6) increased significantly (1.7 times) by d 9 to the same level as in rats administered 50 micromol/kg (6.04 +/- 0.60 micromol/L; n = 6). When VA was removed from the diet on d 7, concentrations declined significantly (by 50%) and by d 11 were the same as those in rats given 0 micromol/kg. We conclude that the rapid effect of changes in dietary VA intake are attributable to changes in the delivery of chylomicron VA to mammary tissue and milk. PMID- 16365072 TI - Branched-chain amino acid needs in children with mild-to-moderate chronic cholestatic liver disease. AB - Protein-energy malnutrition is prevalent in children with chronic cholestatic liver disease. Supplementation of branched-chain amino acids (BCAA) in infants and children with chronic liver disease has been associated with significant improvement in growth and nitrogen balance, suggesting that BCAA requirements are increased in chronic liver disease. The goal of the present study was to determine the total BCAA requirement in children with mild-to-moderate chronic cholestatic (MCC) liver disease using indicator amino acid oxidation (IAAO). Total BCAA requirements were determined in 6 children (6.3 +/- 3.7 y, mean +/- SD) with MCC liver disease. Children were randomly assigned to receive 7 graded intakes of total BCAA. Individual BCAA in the test diet were provided in the same proportions as those present in egg protein to minimize the potential interactive effects of individual BCAA on assessment of requirement. The total BCAA requirement was determined by measuring the oxidation of l-[1-13C] Phe to 13CO2 [F13CO2 in micromol/(kg x h)], after a primed, continuous oral administration of the tracer and using a 2-phase linear regression crossover regression analysis. The estimated mean requirement and the upper limit of the 95% CI for total BCAA establishing using the IAAO in children with MCC liver disease were 209 and 272 mg/(kg x d), respectively. Total BCAA estimated average requirements using the IAAO were significantly higher than mean requirements established previously for healthy children (P < 0.05). PMID- 16365073 TI - Excessive weight gain during pregnancy is associated with earlier termination of breast-feeding among White women. AB - High prepregnant BMI is associated with reduced initiation and duration of breast feeding (BF). To examine how gestational weight gain (GWG) might modify this association, over a 9-y period, we identified all women (n = 2783) who had attempted to breast-feed their newborns. From their medical records, we categorized them by prepregnant BMI [as underweight (<19.8 kg/m2), normal-weight (19.8-26.0 kg/m2), overweight (26.1-29.0 kg/m2) or obese ( >29.0 kg/m2)] and GWG [as below, within, or above the amount recommended by the Institute of Medicine]. Women with a normal BMI who gained within these recommendations served as the reference group in regression analyses, which were adjusted for confounding factors. Both normal-weight (P < 0.05) and obese (P < 0.01) women who exceeded the recommended GWG had higher odds of failing to initiate BF (defined as continuing to breast-feed at 4 d postpartum). Underweight (P < 0.05), overweight (P < 0.05), and obese (P < 0.01) women who exceeded the recommendations for GWG as well as obese women who gained within the recommendations (P < 0.01) had a higher risk of early discontinuation of exclusive BF. Only obese women who gained within or exceeded the recommendations (P < 0.01) for GWG had a higher risk of early discontinuation of any BF. Excessive GWG was associated with a measure of failure to initiate and/or sustain BF in all categories of prepregnant BMI. Thus, in addition to conceiving at a healthy weight, gaining the recommended amount of weight during pregnancy is also important for successful BF. PMID- 16365074 TI - Early programming of body composition and fat distribution in adolescents. AB - Size at birth and early postnatal growth are determinants of adult height and BMI. The aim of this study was to evaluate the effect of birth weight on body composition and fat distribution in a group of Spanish adolescents. Current body composition was assessed by both skinfold thickness and dual X-ray absorptiometry in 234 adolescents born at term (140 girls and 94 boys), now aged 13-18 y and living in the city of Zaragoza. Relative fat distribution was estimated using the ratio of the subscapular to triceps skinfolds (S:T). Birth weight and gestational age were assessed by a questionnaire. Birth weight was inversely associated with the S:T ratio (P < 0.05) in boys and directly associated with bone mass (P < 0.01) and fat-free mass (P < 0.05) in girls. This association was independent of factors such as age, Tanner stage, gestational age, socioeconomic status, physical activity, and height. In conclusion, our data support the hypothesis that impaired fetal growth, measured by birth weight, may be related to central fat distribution in boys and decreased bone and fat-free mass in girls. PMID- 16365075 TI - Trends in serum folate, RBC folate, and circulating total homocysteine concentrations in the United States: analysis of data from National Health and Nutrition Examination Surveys, 1988-1994, 1999-2000, and 2001-2002. AB - Folate intakes increased dramatically after folic acid fortification. We investigated the changes in serum folate, RBC folate, and total homocysteine (tHcy) concentrations utilizing data from National Health and Nutrition Examination Surveys (NHANES) 1988-2002. NHANES 1988-2002 were based on a stratified, multistage, probability sampling design conducted among civilian U.S. residents. The current study included 17,144, 17,213, and 11,415 measurements for serum folate, RBC folate, and tHcy, respectively. Overall, geometric mean serum folate concentrations were 149.6 and 129.8% higher in 1999-2000 and 2001-2002, respectively, than in 1988-1994 (P < 0.0001). Sex-, age-, and race-ethnicity adjusted serum folate was significantly lower in 2001-2002 than in 1999-2000 (10.4%, P < 0.0002). The prevalence of low serum folate decreased from 18.4% in 1988-1994 to 0.8% in 1999-2000 and to 0.2% in 2001-2002 (P < 0.0001). RBC folate increased from 391 nmol/L in 1988-1994 to 618 nmol/L in 1999-2000, and to 611 nmol/L in 2001-2002. Consequently, the prevalence of low RBC folate decreased from 45.8% in 1988-1994 to 7.3% in 1999-2000 and to 7.1% in 2001-2002 (P < 0.0001). Although, RBC folate status improved after folic acid fortification in all race-ethnicities, the prevalence of low RBC folate ( approximately 20.5%) continues to be high in non-Hispanic blacks. Age-, sex-, and race-ethnicity adjusted tHcy declined from 9.5 micromol/L in 1988-1994 to 7.6 mumol/L in 1999 2000 and to 7.9 micromol/L in 2001-2002. Although folic acid fortification contributed to significant improvement in folate status, serum folate concentrations have declined recently. This may be attributable to lower folic acid intakes. PMID- 16365076 TI - Dietary saturated fat intake is inversely associated with bone density in humans: analysis of NHANES III. AB - Mounting evidence indicates that the amount and type of fat in the diet can have important effects on bone health. Most of this evidence is derived from animal studies. Of the few human studies that have been conducted, relatively small numbers of subjects and/or primarily female subjects were included. The present study assessed the relation of dietary fat to hip bone mineral density (BMD) in men and women using NHANES III data (n = 14,850). Multivariate models using SAS callable SUDAAN were used to adjust for the sampling scheme. Models were adjusted for age, sex, weight, height, race, total energy and calcium intakes, smoking, and weight-bearing exercise. Data from women were further adjusted for use of hormone replacement therapy. Including dietary protein, vitamin C, and beta carotene in the model did not influence the outcome. Analysis of covariance was used to generate mean BMD by quintile of total and saturated fat intake for 4 sex/age groups. Saturated fat intake was negatively associated with BMD at several hip sites. The greatest effects were seen among men < 50 y old (linear trend P = 0.004 for the femoral neck). For the femoral neck, adjusted mean BMD was 4.3% less among men with the highest compared with the lowest quintile of saturated fat intake (BMD, 95% CI: highest quintile: 0.922 g/cm2, 0.909-0.935; lowest quintile: 0.963 g/cm2, 95% CI: 0.950-0.976). These data indicate that BMD is negatively associated with saturated fat intake, and that men may be particularly vulnerable to these effects. PMID- 16365077 TI - Consumption of coffee, but not black tea, is associated with decreased risk of premenopausal breast cancer. AB - Caffeine has been suggested as a possible risk factor for breast cancer, potentially through its effect of facilitating the development of benign breast disease. However, coffee and tea also contain polyphenols, which exhibit anticarcinogenic properties. A hospital-based, case-control study was conducted to evaluate the role of coffee, decaffeinated coffee, and black tea in breast cancer etiology. Study participants included 1932 cases with primary, incident breast cancer and 1895 hospital controls with nonneoplastic conditions. All participants completed a comprehensive epidemiological questionnaire. Among premenopausal women, consumption of regular coffee was associated with linear declines in breast cancer risk (P for trend = 0.03); consumers of >or=4 cups/d experienced a 40% risk reduction (odds ratio = 0.62, 95% CI 0.39-0.98). No clear associations between intake of black tea or decaffeinated coffee and breast cancer risk were noted among premenopausal women, although black tea was associated with a protective effect unique to a subsample of cases with lobular histology. Among postmenopausal women, breast cancer risk was not associated with consumption of coffee, tea, or decaffeinated coffee. Results among postmenopausal women did not differ by histologic subtype. Our findings support a protective effect of coffee intake on premenopausal, but not postmenopausal breast cancer risk. Further studies are warranted to confirm these findings. PMID- 16365078 TI - Low serum selenium and total carotenoids predict mortality among older women living in the community: the women's health and aging studies. AB - Selenium and the carotenoids play an important role in antioxidant defenses and in the redox regulation involved in inflammation. We tested the hypothesis that low selenium and carotenoids predict mortality in older women living in the community. Women who were enrolled in the Women's Health and Aging Studies I and II in Baltimore, MD (n = 632; 70-79 y old) had serum selenium and carotenoids measured at baseline and were followed for mortality over 60 mo. Median (minimum, maximum) serum selenium and carotenoids were 1.53 (0.73, 2.51) micromol/L and 1.67 (0.13, 9.10) micromol/L; 14.1% of the women died. The 5 major causes of death were heart disease (32.6%), cancer (18.0%), stroke (9.0%), infection (6.7%), and chronic obstructive pulmonary disease (5.6%). Adjusting for age, education, smoking, BMI, poor appetite, and chronic diseases, higher serum selenium [hazard ratio (HR) 0.71, 95% CI 0.56-0.90/1 SD increase in log(e) selenium; P = 0.005] and higher serum total carotenoids (HR 0.77, 95% CI 0.64 0.84/1 SD increase in log(e) total carotenoids; P = 0.009) were associated with a lower risk of mortality. Women living in the community who have higher serum selenium and carotenoids are at a lower risk of death. PMID- 16365079 TI - Psychosocial factors and socioeconomic indicators are associated with household food insecurity among pregnant women. AB - Household food insecurity has been associated with several negative health outcomes, yet little is known about the prevalence and correlates of household food insecurity during pregnancy. This study was conducted as part of the Pregnancy, Infection, and Nutrition prospective cohort study to identify risk factors of preterm birth. The USDA 18-item scale was used to assess the prevalence of food insecurity among pregnant women with incomes 400 microg/d had reduced NK cytotoxicity compared with those consuming a low-folate diet and no supplements (P = 0.02). Prompted by this observation, we assessed the presence of unmetabolized FA in plasma as a biochemical marker of excess FA. Unmetabolized folic acid was detected in 78% of plasma samples from fasting participants. We found an inverse relation between the presence of unmetabolized FA in plasma and NK cytotoxicity. NK cytotoxicity was approximately 23% lower among women with detectable folic acid (P = 0.04). This inverse relation was stronger among women >or= 60 y old and more pronounced with increasing unmetabolized FA concentrations (P-trend = 0.002). Because of the increased intake of FA in many countries, our findings highlight the need for further studies on the effect of long-term high FA intake on immune function and health. PMID- 16365082 TI - PYY[3-36] administration decreases the respiratory quotient and reduces adiposity in diet-induced obese mice. AB - In rodents, weight reduction after peptide YY[3-36] (PYY[3-36]) administration may be due largely to decreased food consumption. Effects on other processes affecting energy balance (energy expenditure, fuel partitioning, gut nutrient uptake) remain poorly understood. We examined whether s.c. infusion of 1 mg/(kg x d) PYY[3-36] (for up to 7 d) increased metabolic rate, fat combustion, and/or fecal energy loss in obese mice fed a high-fat diet. PYY[3-36] transiently reduced food intake (e.g., 25-43% lower at d 2 relative to pretreatment baseline) and decreased body weight (e.g., 9-10% reduction at d 2 vs. baseline) in 3 separate studies. Mass-specific metabolic rate in kJ/(kg x h) in PYY[3-36] treated mice did not differ from controls. The dark cycle respiratory quotient (RQ) was transiently decreased. On d 2, it was 0.747 +/- 0.008 compared with 0.786 +/- 0.004 for controls (P < 0.001); light cycle RQ was reduced throughout the study in PYY[3-36]-treated mice (0.730 +/- 0.006) compared with controls (0.750 +/- 0.009; P < 0.001). Epididymal fat pad weight in PYY[3-36]-treated mice was approximately 50% lower than in controls (P < 0.01). Fat pad lipolysis ex vivo was not stimulated by PYY[3-36]. PYY[3-36] decreased basal gallbladder emptying in nonobese mice. Fecal energy loss was negligible ( approximately 2% of ingested energy) and did not differ between PYY[3-36]-treated mice and controls. Thus, negative energy balance after PYY[3-36] administration in diet-induced obese mice results from reduced food intake with a relative maintenance of mass specific energy expenditure. Fat loss and reduced RQ highlight the potential for PYY[3-36] to drive increased mobilization of fat stores to help meet energy requirements in this model. PMID- 16365083 TI - In vivo urea kinetic studies in conscious mice. AB - Stable isotope studies in conscious mice have been limited by the invasive catheterization procedures and relatively large sample size required. We developed minimally invasive catheterization protocols that together with the ability to analyze small samples have allowed for the study of urea kinetics in conscious mice. A single dose of 15N15N-urea followed by multiple sampling in mice (n = 6) showed that a primary pool of urea exchanged rapidly [70.65 +/- 14.96 mmol/(kg x h)] with a secondary pool. The urea entry rate determined with this protocol was 3.36 +/- 0.30 mmol/(kg x h). Continuous infusion of 15N15N-urea (n = 6) achieved plateau enrichment values at 3.3 +/- 0.2.h from which the urea entry rate was determined by isotope dilution [3.24 +/- 0.23 mmol/(kg x h)]. The urea entry rate measured by the single dose or continuous infusion protocol did not differ (P = 0.76). The minimally invasive methods described allow us to study not only ureagenesis and urea cycle disorders in vivo, but also urea transport and transporter function and nitrogen metabolism in general in mouse models. This is especially relevant because mouse targeting technologies will likely facilitate the generation of organ and tissue specific nulls of the various urea cycle enzymes. PMID- 16365084 TI - Branched-chain amino acids: enzyme and substrate regulation. AB - The three branched-chain amino acids (BCAAs) are the most hydrophobic of the amino acids and play crucial roles in determining the structures of globular proteins as well as the interaction of the transmembrane domains of membranous proteins with phospholipid bilayers. However, the three BCAAs do not behave identically. In terms of protein secondary structure, valine and isoleucine exhibit a definite preference for the beta-structure, whereas leucine has a higher preference for the alpha-helix. Although mutation of one BCAA to another is commonly regarded as conservative, there are well-documented examples of such substitutions that have a significant effect on protein function. The occurrence of BCAA in nature is, therefore, attributable to their primary role in protein structure, not to their secondary metabolic roles. These functions are important for almost all proteins; therefore, BCAA commonly account for approximately 20 25% of most dietary proteins. Dietary BCAA largely escape first-pass splanchnic metabolism. The first steps in their catabolism are common to all three, involving the BCAA aminotransferase (BCAT) and branched-chain alpha-keto acid dehydrogenase (BCKD). Their further metabolism employs distinct pathways to different end-products (glucose and/or ketone bodies). However, the fact that the flux-generating step for the catabolism of the three BCAAs occurs at one of the common steps indicates that the production of these downstream products are not individually regulated and, hence, may not play important individual roles. The catabolism of the BCAAs is highly regulated by both allosteric and covalent mechanisms. BCKD is inhibited by phosphorylation and activated by dephosphorylation. Allosteric inhibition of the kinase by the branched-chain keto acids (BCKA) (particularly by alpha-ketoisocaproate) serves both as a mechanism for promoting the catabolism of excess quantities of these amino acids as well as for conserving low concentrations of these dietary essential amino acids. Cytosolic and mitochondrial isoenzymes of BCAT have been identified. They are thought to play an important role in brain neurotransmitter metabolism. PMID- 16365085 TI - The regulation of body and skeletal muscle protein metabolism by hormones and amino acids. AB - For many decades, it has been recognized that insulin, growth hormone, glucocorticoids, insulin-like growth factor 1, thyroid hormones, and other hormones regulate body protein metabolism. It has been more recently recognized, but not understood, that humor factors present in states of acute and chronic inflammation could have a strong impact on protein turnover. Most recently, the role of amino acids, acting as signaling molecules, has become increasingly clarified. In aggregate, these factors (together with neuromuscular activity) determine the balance of body protein mass. We will review some of these data, particularly focusing on amino acids, insulin, and the growth hormone axis and their actions in muscle and how these relate to whole-body protein metabolism. PMID- 16365086 TI - Structure of the blood-brain barrier and its role in the transport of amino acids. AB - Brain capillary endothelial cells form the blood-brain barrier (BBB). They are connected by extensive tight junctions, and are polarized into luminal (blood facing) and abluminal (brain-facing) plasma membrane domains. The polar distribution of transport proteins mediates amino acid (AA) homeostasis in the brain. The existence of two facilitative transporters for neutral amino acids (NAAs) on both membranes provides the brain access to essential AAs. Four Na(+) dependent transporters of NAA exist in the abluminal membranes of the BBB. Together these systems have the capability to actively transfer every naturally occurring NAA from the extracellular fluid (ECF) to endothelial cells and from there into circulation. The presence of Na(+)-dependent carriers on the abluminal membrane provides a mechanism by which NAA concentrations in the ECF of brain are maintained at approximately 10% those of the plasma. Also present on the abluminal membrane are at least three Na(+)-dependent systems transporting acidic AAs (EAAT) and a Na(+)-dependent system transporting glutamine (N). Facilitative carriers for glutamine and glutamate are found only in the luminal membrane of the BBB. This organization promotes the net removal of acidic- and nitrogen-rich AAs from the brain and accounts for the low level of glutamate penetration into the central nervous system. The presence of a gamma-glutamyl cycle at the luminal membrane and Na(+)-dependent AA transporters at the abluminal membrane may serve to modulate movement of AAs from blood to the brain. The gamma-glutamyl cycle is expected to generate pyroglutamate (synonymous with oxyproline) within the endothelial cells. Pyroglutamate stimulates secondary active AA transporters at the abluminal membrane, thereby reducing the net influx of AAs to the brain. It is now clear that BBB participates in the active regulation of the AA content of the brain. PMID- 16365087 TI - Signaling pathways and molecular mechanisms through which branched-chain amino acids mediate translational control of protein synthesis. AB - BCAAs stimulate protein synthesis in in vitro preparations of skeletal muscle. Likewise, the stimulation of protein synthesis in skeletal muscle produced by intake of a mixed meal is due largely to BCAAs. Of the three BCAAs, leucine is the one primarily responsible for the stimulation of protein synthesis under these circumstances. The stimulatory effect of leucine on protein synthesis is mediated through upregulation of the initiation of mRNA translation. A number of mechanisms, including phosphorylation of ribosomal protein S6 Kinase, eukaryotic initiation factor (eIF)4E binding protein-1, and eIF4G, contribute to the effect of leucine on translation initiation. These mechanisms not only promote global translation of mRNA but also contribute to processes that mediate discrimination in the selection of mRNA for translation. A key component in a signaling pathway controlling these phosphorylation-induced mechanisms is the protein kinase, termed the mammalian target of rapamycin (mTOR). The activity of mTOR toward downstream targets is controlled in part through its interaction with the regulatory-associated protein of mTOR (known as raptor) and the G protein beta subunit-like protein. Signaling through mTOR is also controlled by upstream members of the pathway such as the Ras homolog enriched in brain (Rheb), a GTPase that activates mTOR, and tuberin (also known as TSC2), a GTPase-activating protein, which, with its binding partner hamartin (also known as TSC1), acts to repress mTOR. Candidates for mediating the action of leucine to stimulate signaling through the mTOR pathway include TSC2, Rheb, and raptor. The current state of our understanding of how leucine acts on these signaling pathways and molecular mechanisms to stimulate protein synthesis in skeletal muscle is summarized in this article. PMID- 16365088 TI - Metabolism and physiological function of branched-chain amino acids: discussion of session 1. PMID- 16365089 TI - Modulations of muscle protein metabolism by branched-chain amino acids in normal and muscle-atrophying rats. AB - It has been shown that BCAAs, especially leucine, regulate skeletal muscle protein metabolism. However, it remains unclear how BCAAs regulate muscle protein metabolism and lead to anabolism in vivo. We examined muscle protein synthesis rate and breakdown rate simultaneously during BCAA infusion in muscle atrophy models as well as in normal healthy rats. Corticosterone-treated rats and hindlimb-immobilized rats were used as muscle atrophy models. Muscle protein synthesis rate and breakdown rate were measured as phenylalanine kinetics across the hindlimb. In anesthetized normal rats, BCAAs stimulated muscle protein synthesis despite low insulin concentration and did not suppress muscle protein breakdown. In corticosterone-treated rats, BCAAs failed to restore inhibited muscle protein synthesis, but reduced muscle protein breakdown. Immobilization of hindlimb increased muscle protein breakdown within a day. BCAAs did not change muscle protein metabolism, although essential amino acids (EAAs) suppressed muscle protein breakdown in hindlimb-immobilized rats. We also evaluated changes of fractional synthesis rate (FSR) of skeletal muscle protein during infusion of leucine alone or EAAs for 4 h in anesthetized normal rats. FSR showed a transient increase at 15-30 min of leucine infusion and then declined, whereas FSR stayed elevated throughout EAA infusion. We concluded that 1) BCAAs primarily stimulate muscle protein synthesis in normal rats independently of insulin; 2) EAAs are required to maintain the BCAA stimulation of muscle protein synthesis; and 3) The effects of BCAAs on muscle protein metabolism differ between atrophy models. PMID- 16365090 TI - Investigations of branched-chain amino acids and their metabolites in animal models of cancer. AB - Many of the features of BCAA metabolism in the tumor-bearing state are similar to the other disease states that feature involuntary weight loss and skeletal muscle atrophy. These states are generally characterized by altered BCAA availability (low BCAA intakes, elevated rates of BCAA oxidation, and gluconeogenesis), which are concurrent with activation of proteolysis and suppression of protein synthesis in skeletal muscle and ultimately lead to erosion of lean tissue mass. These features in turn imply BCAA deficiency compared with whole-body requirements and are the basis of suggestions for dietary supplementation with BCAA or their metabolites. Recent studies on BCAA supplementation in cancer focus on leucine and its derivative, beta-methyl beta-hydroxybutyrate, as regulators of skeletal muscle metabolism, although their relative efficacy is unknown. However, what would otherwise be a relatively straightforward consideration of amino acid supply and demand is confounded by the presence of the tumor and its potential utilization of BCAA for its proliferative and invasive activities. Positron emission tomography with (11)C-leucine, used for in vivo tumor imaging, points to the high avidity of tumor amino acid uptake. These features have incited research in opposing directions, probing BCAA deprivation, with a view to limiting tumor growth, as well as BCAA supplementation, with a view to supporting maintenance of host lean tissue. No clear conclusion is presently available from the sum of these efforts. Animal models with relevant clinical features are essential to determine if amino acid therapy can alter the balance between the host and the tumor in a manner that favors the host overall. PMID- 16365091 TI - Lessons from genetic disorders of branched-chain amino acid metabolism. AB - Genetic disorders of BCAA metabolism produce amino acidopathies and various forms of organic aciduria with severe clinical consequences. A metabolic block in the oxidative decarboxylation of BCAA caused by mutations in the mitochondrial branched-chain alpha-keto acid dehydrogenase complex (BCKDC) results in Maple Syrup Urine Disease (MSUD) or branched-chain ketoaciduria. There are presently five known clinical phenotypes for MSUD, i.e., classic, intermediate, intermittent, thiamin-responsive, and dihydrolipoamide dehydrogenase (E3) deficient, based on severity of the disease, response to thiamin therapy, and the gene locus affected. Reduced glutamate, glutamine, and gamma-aminobutyrate concentrations induced by the accumulation of branched-chain alpha-ketoacids in the brain cortex of affected children and neonatal polled Hereford calves are considered the cause of MSUD encephalopathies. The long-term restriction of BCAA intake in diets and orthotopic liver transplantation have proven effective in controlling plasma BCAA levels and mitigating some of the above neurological manifestations. To date, approximately 100 mutations have been identified in four (branched-chain alpha-ketoacid decarboxylase/dehydrogenasealpha [E1alpha], E1beta, dihydrolipoyl transacylase [E2], and E3) of the six genes that encode the human BCKDC catalytic machine. We have documented a strong correlation between the presence of mutant E2 proteins and the thiamin-responsive MSUD phenotype. We show that the normal E1 component possesses residual decarboxylase activity, which is augmented by the binding to a mutant E2 protein in the presence of the E1 cofactor thiamin diphosphate. Our results provide a biochemical model for the effectiveness of thiamin therapy to thiamin-responsive MSUD patients. PMID- 16365093 TI - Application of branched-chain amino acids in experimental animals: discussion of session 2. PMID- 16365092 TI - Branched-chain amino acid catabolism in exercise and liver disease. AB - Branched-chain alpha-keto acid dehydrogenase (BCKDH) complex, the enzyme catalyst for the second step of the BCAA catabolic pathway, plays a central role in the regulation of BCAA catabolism. The activity of the complex is regulated by a covalent modification cycle in which phosphorylation by BCKDH kinase inactivates and dephosphorylation by BCKDH phosphatase activates the complex. Many studies suggest that control of the activity of the kinase is a primary determinant of the activity of the complex. The kinase exists at all times in the mitochondrial matrix space in two forms, with a large amount being free and a smaller amount bound rather tightly to the BCKDH complex. Only the bound form of the kinase appears to be catalytically active and, therefore, responsible for phosphorylation and inactivation of the complex. alpha-Ketoisocaproate, the transamination product of leucine and the most important known physiological inhibitor of BCKDH kinase, promotes release of the kinase from the complex. alpha Chloroisocaproate, the analogue of leucine and the most potent known inhibitor of the kinase, is more effective than alpha-ketoisocaproate in promoting release of BCKDH kinase from the complex. Exercise and chronic liver disease (liver cirrhosis) likewise decrease the amount of the kinase bound to the complex in rat liver. The resulting activation of the BCKDH complex appears responsible for the increase in BCAA catabolism caused by exercise and liver cirrhosis. Our findings support the use of BCAA supplements for patients with liver cirrhosis. PMID- 16365094 TI - Branched-chain amino acid requirements in healthy adult human subjects. AB - There is now an expanding body of evidence to recommend, in the case of adult humans, the use of revised indispensable amino acid requirement values; these are approximately 2 to 3 times higher than the current international recommendations. The earlier methodologies for determining amino acid requirements, based on nitrogen balance, were criticized because of their design and the associated high energy intakes. The 1985 World Health Organization/Food & Agriculture Organization/United Nations University requirement for leucine has been demonstrated to be too low by short- and long-term (24-h) tracer-derived estimates of leucine oxidation and balance. The best values for leucine requirements come from 24-h direct amino acid oxidation (DAAO) and direct amino acid balance (DAAB) studies. Finally, we also collated all available data from studies on fed-state leucine oxidation with an adequate dietary adaptation period to assess the inflection on the leucine oxidation-leucine intake curve. The mean requirements for leucine, valine, and isoleucine are likely to be 40, 17-25, and 19 mg . kg(-1) . d(-1), respectively. This adds up to a total of approximately 84 mg . kg(-1) . d(-1), which is much lower than the lowest estimate of the total BCAA requirement of approximately 110 mg . kg(-1) . d(-1) made by the short-term indicator amino acid oxidation (IAAO) method, which determined the BCAA requirement from the pattern of oxidation of an indicator amino acid (phenylyalanine) at different levels of BCAA intake. An additional estimate of the leucine requirement was also made by a meta-analysis of all available 24-h DAAO/DAAB data from different studies. This resulted in a higher value for the leucine requirement than that obtained by the specific studies that utilized the 24-h DAAO/DAAB approach; however, even adding this value to the total BCAA requirement does not account for the difference in the total BCAA requirement estimates and the summed individual BCAA estimates. PMID- 16365095 TI - Branched-chain amino acids as fuels and anabolic signals in human muscle. AB - During exercise, there is an increase in amino acid (AA) oxidation accompanied by a depression in whole-body protein synthesis and an increase in protein breakdown. Leucine oxidation increases in proportion to energy expenditure, but the total contribution of BCAA to fuel provision during exercise is minor and insufficient to increase dietary protein requirements. When investigating the effects of AA on the control of muscle protein synthesis (MPS), we showed that increased availability of mixed AAs caused a rise in human MPS to about the same extent as complete meals. Leucine alone (and to some extent other essential, but not nonessential, AAs) can stimulate MPS for a short period, suggesting that leucine acts as a signal as well as a substrate. MPS stimulation by infused AAs shows tachyphylaxis, returning to basal rates after 2 h, possibly explaining why chronically elevated leucine delivery does not elevate MPS clinically. Increased availability of essential amino acids (EAAs) results in dose-related responses of MPS, but, in elderly subjects, there is blunted sensitivity and responsiveness associated with decreased total RNA and mRNA for signaling proteins and signaling activity. Increases of MPS due to EAAs are associated with elevation of signaling activity in the mammalian target of rapamycin (mTOR)/p70 ribosomal subunit S6 kinase eukaryotic initiation factor 4 binding protein 1 pathway, without requiring rises of plasma insulin availability above 10 microU/mL. However, at insulin of <5 microU/mL, AAs appear to stimulate MPS without increasing mTOR signaling. Further increasing availability of insulin to postprandial values increases signaling activity, but has no further effect on MPS. PMID- 16365096 TI - Branched-chain amino acids activate key enzymes in protein synthesis after physical exercise. AB - BCAAs (leucine, isoleucine, and valine), particularly leucine, have anabolic effects on protein metabolism by increasing the rate of protein synthesis and decreasing the rate of protein degradation in resting human muscle. Also, during recovery from endurance exercise, BCAAs were found to have anabolic effects in human muscle. These effects are likely to be mediated through changes in signaling pathways controlling protein synthesis. This involves phosphorylation of the mammalian target of rapamycin (mTOR) and sequential activation of 70-kD S6 protein kinase (p70 S6 kinase) and the eukaryotic initiation factor 4E-binding protein 1. Activation of p70 S6 kinase, and subsequent phopsphorylation of the ribosomal protein S6, is associated with enhanced translation of specific mRNAs. When BCAAs were supplied to subjects during and after one session of quadriceps muscle resistance exercise, an increase in mTOR, p70 S6 kinase, and S6 phosphorylation was found in the recovery period after the exercise with no effect of BCAAs on Akt or glycogen synthase kinase 3 (GSK-3) phosphorylation. Exercise without BCAA intake led to a partial phosphorylation of p70 S6 kinase without activating the enzyme, a decrease in Akt phosphorylation, and no change in GSK-3. It has previously been shown that leucine infusion increases p70 S6 kinase phosphorylation in an Akt-independent manner in resting subjects; however, a relation between mTOR and p70 S6 kinase has not been reported previously. The results suggest that BCAAs activate mTOR and p70 S6 kinase in human muscle in the recovery period after exercise and that GSK-3 is not involved in the anabolic action of BCAAs on human muscle. PMID- 16365097 TI - Branched-chain amino acids and central fatigue. AB - An account of the tryptophan (Trp)-5-hydroxytryptamine (5-HT)-central fatigue theory is provided and an explanation of how oral administration of BCAAs can decrease fatigue on the basis of this theory is given. The rate-limiting step in the synthesis of 5-HT is the transport of Trp across the blood-brain barrier. This transport is influenced by the fraction of Trp available for transport into the brain and the concentration of the other large neutral amino acids, including the BCAAs, which are transported via the same carrier system. During endurance exercise, there is an uptake of Trp by the brain, suggesting that this may increase the synthesis and release of 5-HT in the brain. Oral intake of BCAAs may reduce this uptake and also brain 5-HT synthesis and release, thereby delaying fatigue. Other hypotheses for the effect of BCAAs on central fatigue are included. PMID- 16365098 TI - Amino acids and muscle loss with aging. AB - Aging is associated with a progressive loss of muscle mass (sarcopenia), which increases the risks of injury and disability. Although the mechanisms of sarcopenia are not clearly elucidated, age-associated alterations in the muscle anabolic response to nutritional stimuli and a decline in protein intake may be significant contributing factors. The most recent findings regarding the role of nutritional intake on protein metabolism in the elderly will be reviewed. Specifically, aging is associated with changes in the muscle protein metabolism response to a meal, likely due to alterations in the response to endogenous hormones. Nonetheless, the older muscle is still able to respond to amino acids, mainly the essential and BCAAs, which have been shown to acutely stimulate muscle protein synthesis in older individuals. It is likely that this stimulatory effect of essential and BCAA is due to the direct effect of leucine on the initiation of mRNA translation, which is still present in older age, although it appears to be attenuated in aged animals. Recent data suggest that excess leucine may be able to overcome this age-related resistance of muscle proteins to leucine. For this reason, long-term essential amino acid supplementation may be a useful tool for the prevention and treatment of sarcopenia, particularly if excess leucine is provided in the supplement. PMID- 16365099 TI - Transamination of leucine and nitrogen accretion in human pregnancy and the newborn infant. AB - Kinetics of leucine and its oxidation were determined in human pregnancy and in the newborn infant, using stable isotopic tracers, to quantify the dynamic aspects of protein metabolism. These data show that in human pregnancy there is a decrease in whole-body rate of leucine turnover compared with nonpregnant women. In addition, data in newborn infants show that leucine turnover expressed as per kg body weight is higher compared with adults. The administering of nutrients resulted in a suppression of the whole-body rate of proteolysis. Because nonessential amino nitrogen is an important component of nutritional nitrogen and can be limiting for growth under certain circumstances, and because BCAA are an important source of nonessential amino nitrogen, we have examined the relations among the transamination of leucine, leucine N kinetics, and urea synthesis and glutamine kinetics in human pregnancy and newborn infants. In human pregnancy, early in gestation, there is a significant decrease in urea synthesis in association with a decrease in the rate of transamination of leucine. A linear correlation was evident between the rate of leucine reamination and urea synthesis during fasting in pregnant and nonpregnant women. In healthy-term newborn and growing infants, although the reamination of leucine was positively related to glutamine flux, leucine reamination was negatively related to urea synthesis, suggesting a redirection of amino N toward protein accretion. The regulatory mechanism involved in this redirection of nitrogen from irreversible loss to accretion remains under investigation. PMID- 16365100 TI - Branched-chain amino acids and immunity. AB - Although there has been great interest in the effects of amino acids on immune function, little is known about the impact of changes in BCAA availability on the ability of the immune system to function. Human immune cells incorporate BCAA into proteins and are able to oxidize BCAA. The immune system exists to protect the host from pathogenic invaders and from other noxious insults. Upon infection, there is a marked increase in demand for substrates by the immune system; these substrates provide energy and are the precursors for the synthesis of new cells, effector molecules, and protective molecules. Cell culture studies show that BCAA are absolutely essential for lymphocytes to synthesize protein, RNA, and DNA and to divide in response to stimulation. In mice, dietary BCAA restriction impairs several aspects of the immune function and increases the susceptibility to pathogens. Postsurgical or septic patients given BCAA intravenously showed improved immunity and this may relate to improved outcome. BCAAs are therefore absolutely essential for lymphocyte responsiveness and are necessary to support other immune cell functions. However, many aspects of BCAA and its effects on immune function have been understudied or not studied at all. More research is needed to understand the extent of the immune system's requirement for BCAA. It is likely that the essentiality of BCAA for the function of immune cells relates to protein synthesis. PMID- 16365101 TI - Application of branched-chain amino acids in healthy humans: discussion of session 3. PMID- 16365102 TI - Branched-chain amino acid enriched supplements as therapy for liver disease. AB - Altered amino acid metabolism is a hallmark of liver disease, characterized by low levels of circulating BCAAs and elevated levels of circulating aromatic amino acids, and methionine. Although overwhelming evidence indicates that the incidence of complications of liver disease increases with malnutrition, the reported impact of nutritional therapy, specifically BCAA supplementation, on outcomes in patients with liver disease has varied with the indication. Multiple small studies report the beneficial effects of BCAA supplementation, including improved metabolic profiles, as measured by protein sparing and/or normalization of respiratory quotients and clinical improvement of hepatic encephalopathy. Other studies have failed to show a clinical benefit of BCAA supplementation. The data concerning the impact of BCAA supplementation in prophylaxis of long-term morbidity and mortality in patients with cirrhosis is more promising and has been the subject of 2, large randomized controlled trials. In a study of 174 patients with advanced cirrhosis, who were randomized to either BCAA or 1 of 2 control arms, the combined event rates were seen to be significantly reduced in the BCAA supplementation arm, although this was not true for individual complications. In a more recent, larger, randomized controlled trial (n = 646) using a more palatable formulation, investigators demonstrated that long-term BCAA supplementation is associated with decreased frequency of hepatic failure and overall complication frequency. Both studies found improved nutritional status associated with BCAA supplementation. On balance, BCAA supplementation appears to be associated with decreased frequency of complications of cirrhosis and improved nutritional status when prescribed as maintenance therapy. Cost and palatability may limit the potential applicability of this treatment modality. PMID- 16365103 TI - Application of branched-chain amino acids in human pathological states: renal failure. AB - During renal failure, abnormalities of BCAA and branched-chain keto acid (BCKA) metabolism are due to both the lack of renal contribution to amino acid metabolism and the impact of renal failure and acidosis on whole-body nitrogen metabolism. Abnormal BCAA and BCKA metabolism result in BCAA depletion as reflected by low plasma BCAAs and cellular valine. BCAA metabolic disturbances can alter tissue activities, particularly brain function, and nutritional status. In dialysis patients, BCAA oral supplementation can induce an improvement of appetite and nutritional status. During chronic renal failure, the aims of nutritional interventions are to minimize uremic toxicity, avoid malnutrition and delay progression of kidney disease. BCAA and BCKA supplements have been proposed to decrease further protein intake while maintaining satisfactory nutritional status. In this setting, BCAAs or BCKAs have not been administrated solely but in association with other essential AA or keto analogs. Therefore, the proper effects of BCAAs and/or BCKAs have not been studied separately. Protein restriction together with keto acids and/or essential AAs has been reported to improve insulin sensitivity and hyperparathyroidism and to be compatible with a preservation of nutritional status. Nonetheless, a careful monitoring of protein calorie intake and nutritional status is needed. A recent meta-analysis concluded that reducing protein intake in patients with chronic renal failure reduces the occurrence of renal death by approximately 40% as compared with larger or unrestricted protein intake. The additional effect of essential amino acids and keto acids on retardation of progression of renal failure has not been demonstrated. PMID- 16365104 TI - Therapeutic use of branched-chain amino acids in burn, trauma, and sepsis. AB - Various experimental studies conducted in the 1970s demonstrated, at least in the physiological situation, the anabolic and/or anticatabolic properties of branched chain amino acids (leucine, valine, isoleucine) or their ketoacid derivatives. This led to several clinical studies in the late 1970s and early 1980s that aimed to evaluate the potential benefits of BCAA supplementation in nutritional support of the critically ill. The data on burn, trauma, and sepsis are, however, far from convincing. Besides significant discrepancies in their results and the fact that most of these studies involved very small populations of patients, few of them meet the current standards of therapeutic evaluation. However, some positive results in specific studies suggest that the underlying concept may be correct but that interpretation has been faulty. Indeed, we know now that while the BCAAs possess regulatory properties on protein metabolism, leucine is by far the most potent, while isoleucine and valine are inefficient. However, in the above mentioned studies, BCAA-supplemented nutrition very frequently supplied almost equivalent amounts of all 3 BCAAs. Moreover, several studies were performed without adequate basal nutritional support, which most probably hampered the correct metabolic utilization of these amino acids. Taken together, these factors mean that the demonstrations of BCAA efficacy were fortunate in the least. In contrast, more recently, leucine was demonstrated to positively affect protein synthesis in an experimental model of sepsis or burn. In parallel, 2 prospective controlled trials of BCAA supplementation in septic patients also demonstrated an improvement in patients' nutritional status and outcome. Thus, we should abandon the concept of BCAA-supplemented nutrition for a more promising leucine supplemented nutrition that requires further evaluation. PMID- 16365105 TI - Branched-chain amino acid-enriched nutritional support in surgical and cancer patients. AB - Prolonged surgical stress and advanced malignant disease lead to systemic catabolism characterized by depletion of muscle protein and oxidation of skeletal muscle BCAA. BCAA oxidation provides energy for muscle and other organs and is the precursor for amino acid synthesis to replenish alanine and glutamine depleted in catabolic states. Persistent excessive catabolism leads to skeletal muscle wasting, negative nitrogen balance, and immune compromise. BCAAs, especially leucine, stimulate protein synthesis, inhibit proteolysis (in cell culture models and in animals), and promote glutamine synthesis. A number of small and diverse clinical trials studied the effects of BCAA-enriched nutritional support in moderately to severely stressed surgical and cancer patients. The findings of these clinical trials have been inconsistent; some show improved nitrogen balance, increased skeletal muscle protein synthesis, and reduced skeletal muscle catabolism whereas others show no significant improvement. The value of these trials is compromised by small sample size, heterogeneous patients, poor study design, varying degrees of metabolic stress, and inappropriate endpoints. More recent trials that evaluate clinical outcomes in hepatocellular carcinoma patients show promising results; in addition to improving metabolic parameters, BCAA-enriched oral supplementation improved morbidity and quality of life in patients undergoing major liver resection and chemo-embolization. In summary, the role of BCAAs in the nutritional support of stressed surgical and cancer patients remains to be clearly defined, despite their potential beneficial biological properties. PMID- 16365106 TI - Potential importance of leucine in treatment of obesity and the metabolic syndrome. AB - Diets with total protein intake >1.5 g.kg(-1).d(-1) and carbohydrate intake <150 g/d are effective for treatment of obesity, type 2 diabetes, and the Metabolic Syndrome. These diets improve body composition and enhance glycemic control. During weight loss, protein-rich diets reduce loss of lean tissue and increase loss of body fat. Specific mechanisms to explain each of these clinical outcomes remain to be fully elucidated. We propose that keys to understanding the relationship between dietary protein and carbohydrates are the relationships between the branched-chain amino acid leucine and insulin and glucose metabolism. Leucine is known to interact with the insulin signaling pathway to stimulate downstream signal control of protein synthesis, resulting in maintenance of muscle protein during periods of restricted energy intake. Leucine also appears to modulate insulin signaling and glucose use by skeletal muscle. Whereas total protein is important in providing substrates for gluconeogenesis, leucine appears to regulate oxidative use of glucose by skeletal muscle through stimulation of glucose recycling via the glucose-alanine cycle. These mechanisms produce protein sparing and provide a stable glucose environment with low insulin responses during energy-restricted periods. PMID- 16365107 TI - Assessment of branched-chain amino Acid status and potential for biomarkers. AB - BCAAs are not synthesized in the body in humans, but they are crucial in protein and neurotransmitter synthesis. The protein anabolic role of BCAAs seems to be mediated not only by their important role as a promoter of the translation process (and possibly acting at the transcription level) but also by inhibition of protein degradation. Leucine may play a critical role in these signaling pathways. Supplementation with BCAAs spares lean body mass during weight loss, promotes wound healing, may decrease muscle wasting with aging, and may have beneficial effects in renal and liver disease. BCAA supplementation is extensively used in the athletic field with the assumption of improved performance and muscle mass. Measuring serum BCAAs has limited clinical utility beyond the controlled setting because levels are affected by a variety of clinical states, and optimal levels in these scenarios have not been completely elucidated. We discuss the effects diet, hormones, stress, aging, and renal or liver dysfunction have on BCAA levels and how understanding the biological effects of BCAAs may help to develop biomarkers of BCAA status. We also discuss potential biomarkers of BCAA status. PMID- 16365108 TI - Application of branched-chain amino acids in human pathological states: discussion of session 4. PMID- 16365109 TI - Symposium on branched-chain amino acids: conference summary. PMID- 16365111 TI - Informed consent--a contemporary myth? PMID- 16365112 TI - The management of complex soft-tissue defects after spinal instrumentation. PMID- 16365113 TI - Consent--an event or a memory? A judicial view. PMID- 16365114 TI - CT analysis of defects of the cement mantle and alignment of the stem: in vitro comparison of Charnley-Kerboul femoral hip implants inserted line-to-line and undersized in paired femora. AB - Using a modern cementing technique, we implanted 22 stereolithographic polymeric replicas of the Charnley-Kerboul stem in 11 pairs of human cadaver femora. On one side, the replicas were cemented line-to-line with the largest broach. On the other, one-size undersized replicas were used (radial difference, 0.89 mm sd 0.13).CT analysis showed that the line-to-line stems without distal centralisers were at least as well aligned and centered as undersized stems with a centraliser, but were surrounded by less cement and presented more areas of thin (< 2 mm) or deficient (< 1 mm) cement. These areas were located predominantly at the corners and in the middle and distal thirds of the stem. Nevertheless, in line-to-line stems, penetration of cement into cancellous bone resulted in a mean thickness of cement of 3.1 mm (sd 0.6) and only 6.2% of deficient and 26.4% of thin cement. In over 90% of these areas, the cement was directly supported by cortical bone or cortical bone with less than 1 mm of cancellous bone interposed. When Charnley-Kerboul stems are cemented line-to-line, good clinical results are observed because cement-deficient areas are limited and are frequently supported by cortical bone. PMID- 16365115 TI - Risk factors for failure after treatment of a periprosthetic fracture of the femur. AB - Periprosthetic fracture of the femur is an uncommon complication after total hip replacement, but appears to be increasing. We undertook a nationwide observational study to determine the risk factors for failure after treatment of these fractures, examining patient- and implant-related factors, the classification of the fractures and the outcome. Between 1979 and 2000, 1049 periprosthetic fractures of the femur were reported to the Swedish National Hip Arthroplasty Register. Of these, 245 had a further operation after failure of their initial management. Data were collected from the Register and hospital records. The material was analysed by the use of Poisson regression models. It was found that the risk of failure of treatment was reduced for Vancouver type B2 injuries (p = 0.0053) if revision of the implant was undertaken (p = 0.0033) or revision and open reduction and internal fixation (p = 0.0039) were performed. Fractures classified as Vancouver type B1 had a significantly higher risk of failure (p = 0.0001). The strongest negative factor was the use of a single plate for fixation (p = 0.001). The most common reasons for failure in this group were loosening of the femoral prosthesis, nonunion and re-fracture. It is probable that many fractures classified as Vancouver type B1 (n = 304), were in reality type B2 fractures with a loose stem which were not recognised. Plate fixation was inadequate in these cases. The difficulty in separating type B1 from type B2 fractures suggests that the prosthesis should be considered as loose until proven otherwise. PMID- 16365116 TI - Removal of acetabular bone in resurfacing arthroplasty of the hip: a comparison with hybrid total hip arthroplasty. AB - Resurfacing arthroplasty of the hip is being performed more frequently in the United Kingdom. The majority of these patients are younger than 55 years of age, and in this group the key benefits include conservation of femoral bone stock and the potential reduction in the rate of dislocation afforded by the larger resurfacing head. Early aseptic loosening is well recognised in patients younger than 55 years of age, and proponents of resurfacing believe that the improved wear characteristics of the metal-on-metal bearing may improve the long-term survival of this implant. There has been some concern, however, that resurfacing may not be conservative of acetabular bone. We compared a series of 33 consecutive patients who had a hybrid total hip arthroplasty with an uncemented acetabular component and a cemented femoral implant, with 35 patients undergoing a Birmingham hip resurfacing arthroplasty. We compared the diameter of the implanted acetabulum in both groups and, because they were not directly comparable, we corrected for patient size by measuring the diameter of the contralateral femoral head. The data were analysed using unpaired t-tests and analysis of covariance. There was a significantly larger acetabulum in the Birmingham arthroplasty group (mean diameter 56.6 mm vs 52.0 mm; p < 0.001). However, this group had a significantly larger femoral head diameter on the contralateral side (p = 0.03). Analysis of covariance revealed a significant difference between the mean size of the acetabular component implanted in the two operations. The greatest difference in the size of acetabulum was in those patients with a larger diameter of the femoral head. This study shows that more bone is removed from the acetabulum in hip resurfacing than during hybrid total hip arthroplasty, a difference which is most marked in larger patients. PMID- 16365117 TI - Notching of the femoral neck during resurfacing arthroplasty of the hip: a vascular study. AB - During hip resurfacing arthroplasty, excessive valgus positioning or surgical technique can result in notching of the femoral neck. Although mechanical weakening and subsequent fracture of the femoral neck are well described, the potential damage to the retinacular vessels leading to an ischaemic event is relatively unknown. Using laser Doppler flowmetry, we measured the blood flow in 14 osteoarthritic femoral heads during routine total hip replacement surgery, before and after notching of the femoral neck. In ten hips there was a reduction in blood flow of more than 50% from the baseline value after simulated notching of the femoral neck. Our results suggest that femoral head vascularity in the osteoarthritic state is similar to the non-arthritic state, where damage to the extraosseous vessels can predispose to avascular necrosis. Surgeons who perform resurfacing arthroplasty of the hip should pay careful attention to these vessels by avoiding excessive dissection around the femoral neck and/or notching. PMID- 16365118 TI - A comparison of bilateral uncemented total knee arthroplasty: simultaneous or staged? AB - In a series of 1304 patients (1867 knees), the results of simultaneous and staged bilateral total knee arthroplasty were compared with each other and with unilateral total knee arthroplasty. The bilateral procedures had a significantly higher rate of complications than unilateral procedures, almost entirely because of thromboembolic problems. However, this did not correspond to an increase in mortality. If a bilateral procedure was indicated, then a simultaneous procedure had no increased risk over a staged procedure. There was no increase in cardiovascular complications, the rate of deep-vein thrombosis or pulmonary embolism or mortality. The rate of infection was lower with a bilateral procedure and the overall revision rate was less than 1% in all groups. The prosthesis functioned as well in all groups in the medium and longer term periods. We feel that simultaneous bilateral total knee arthroplasty is a safe and successful procedure when compared with a staged bilateral procedure. It also has the added benefit of single anaesthetic, reduced costs and decreased total recovery time when compared to a staged bilateral procedure. For these reasons it should be considered as an option in the presence of bilateral knee joint disease. PMID- 16365119 TI - Limb alignment in computer-assisted minimally-invasive unicompartmental knee replacement. AB - Twenty patients underwent simultaneous bilateral medial unicompartmental knee arthroplasty. Pre-operative hip-knee-ankle alignment and valgus stress radiographs were used to plan the desired post-operative alignment of the limb in accordance with established principles for unicompartmental arthroplasty. In each patient the planned alignment was the same for both knees. Overall, the mean planned post-operative alignment was to 2.3 degrees of varus (0 degrees to 5 degrees ). The side and starting order of surgery were randomised, using conventional instrumentation for one knee and computer-assisted surgery for the opposite side. The mean variation between the pre-operative plan and the achieved correction in the navigated and the non-navigated limb was 0.9 degrees (sd 1.1; 0 degrees to 4 degrees ) and 2.8 degrees (sd 1.4; 1 degrees to 7 degrees ), respectively. Using the Wilcoxon signed rank test, we found the difference in variation statistically significant (p < 0.001). Assessment of lower limb alignment in the non-navigated group revealed that 12 (60%) were within +/- 2 degrees of the pre-operative plan, compared to 17 (87%) of the navigated cases.Computer-assisted surgery significantly improves the post-operative alignment of medial unicompartmental knee arthroplasty compared to conventional techniques in patients undergoing bilateral simultaneous arthroplasty. Improved alignment after arthroplasty is associated with better function and increased longevity. PMID- 16365120 TI - A prospective comparison of clamping the drain or post-operative salvage of blood in reducing blood loss after total knee arthroplasty. AB - We undertook a prospective, randomised study in order to evaluate the efficacy of clamping the drains after intra-articular injection of saline with 1:500 000 adrenaline compared with post-operative blood salvage in reducing blood loss in 212 total knee arthroplasties. The mean post-operative drained blood volume after drain clamping was 352.1 ml compared to 662.3 ml after blood salvage (p < 0.0001). Allogenic blood transfusion was needed in one patient in the drain group and for three in the blood salvage group. Drain clamping with intra-articular injection of saline with adrenaline is more effective than post-operative autologous blood transfusion in reducing blood loss during total knee arthroplasty. PMID- 16365121 TI - The Oxford medial unicompartmental knee replacement using a minimally-invasive approach. AB - This prospective study describes the complications and survival of the first 688 Phase 3 Oxford medial unicompartmental knee replacements implanted using a minimally-invasive technique by two surgeons and followed up independently. None was lost to follow-up. We had carried out 132 of the procedures more than five years ago. The clinical assessment of 101 of these which were available for review at five years is also presented. Nine of the 688 knees were revised: four for infection, three for dislocation of the bearing and two for unexplained pain. A further seven knees (1%) required other procedures: four had a manipulation under anaesthesia, two an arthroscopy and one a debridement for superficial infection. The survival rate at seven years was 97.3% (95% confidence interval 5.3). At five years, 96% of the patients had a good or excellent American Knee Society score, the mean Oxford knee score was 39 and the mean flexion was 133 degrees. This study demonstrates that the minimally-invasive Oxford unicompartmental knee replacement is a reliable and effective procedure. PMID- 16365122 TI - Who is the ideal candidate for autologous chondrocyte implantation? AB - We investigated the prognostic indicators for collagen-covered autologous chondrocyte implantation (ACI-C) performed for symptomatic osteochondral defects of the knee. We analysed prospectively 199 patients for up to four years after surgery using the modified Cincinnati score. Arthroscopic assessment and biopsy of the neocartilage was also performed whenever possible. The favourable factors for ACI-C include younger patients with higher pre-operative modified Cincinnati scores, a less than two-year history of symptoms, a single defect, a defect on the trochlea or lateral femoral condyle and patients with fewer than two previous procedures on the index knee. Revision ACI-C in patients with previous ACI and mosaicplasties which had failed produced significantly inferior clinical results. Gender (p = 0.20) and the size of the defect (p = 0.97) did not significantly influence the outcome. PMID- 16365123 TI - An equinus deformity of the ankle accounts for only a small amount of the increased forefoot plantar pressure in patients with diabetes. AB - Patients with diabetes mellitus may develop plantar flexion contractures (equinus) which may increase forefoot pressure during walking. In order to determine the relationship between equinus and forefoot pressure, we measured forefoot pressure during walking in 27 adult diabetics with a mean age of 66.3 years (sd 7.4) and a mean duration of the condition of 13.4 years (sd 12.6) using an Emed mat. Maximum dorsiflexion of the ankle was determined using a custom device which an examiner used to apply a dorsiflexing torque of 10 Nm (sd 1) for five seconds. Simple linear regression showed that the relationship between equinus and peak forefoot pressure was significant (p < 0.0471), but that only a small portion of the variance was accounted for (R(2) = 0.149). This indicates that equinus has only a limited role in causing high forefoot pressure. Our findings suggest caution in undertaking of tendon-lengthening procedures to reduce peak forefoot plantar pressures in diabetic subjects until clearer indications are established. PMID- 16365124 TI - Randomised controlled trials of immediate weight-bearing mobilisation for rupture of the tendo Achillis. AB - We performed two independent, randomised, controlled trials in order to assess the potential benefits of immediate weight-bearing mobilisation after rupture of the tendo Achillis. The first trial, on operatively-treated patients showed an improved functional outcome for patients mobilised fully weight-bearing after surgical repair. Two cases of re-rupture in the treatment group suggested that careful patient selection is required as patients need to follow a structured rehabilitation regimen. The second trial, on conservatively-treated patients, provided no evidence of a functional benefit from immediate weight-bearing mobilisation. However, the practical advantages of immediate weight-bearing did not predispose the patients to a higher complication rate. In particular, there was no evidence of tendon lengthening or a higher re-rupture rate. We would advocate immediate weight-bearing mobilisation for the rehabilitation of all patients with rupture of the tendo Achillis. PMID- 16365125 TI - Revision of failed total elbow arthroplasty with use of a linked implant. AB - We studied retrospectively the results of revision arthroplasty of the elbow using a linked Coonrad-Morrey implant in 23 patients (24 elbows) after a mean follow-up period of 55 months. According to the Mayo Elbow Performance Score, 19 elbows were satisfactory, nine were excellent and ten good. The median total score had improved from 35 points (20 to 75) before the primary arthroplasty to 85 points (40 to 100) at the latest follow-up. There was a marked relief of pain, but the range of movement showed no overall improvement. Two patients had a second revision because of infection and two for aseptic loosening. The estimated five-year survival rate of the prosthesis was 83.1% (95% confidence interval 61.1 to 93.3). Revision elbow arthroplasty using the Coonrad-Morrey implant provided satisfactory results but with complications occurring in 13 cases. PMID- 16365126 TI - The posterior Monteggia lesion with associated ulnohumeral instability. AB - The type II Monteggia (posterior) lesion is a rare injury which is sometimes associated with ulnohumeral instability. We have reviewed 23 of 28 patients with this injury. A clinical and radiographic assessment was undertaken at follow-up. Functional outcome scores, including the Broberg and Morrey Index and the Disabilities of the Arm, Shoulder or Hand (DASH), were used. The results from the six patients with associated posterior ulnohumeral dislocation were compared with 17 without ulnohumeral injury. Those with dislocation had reduced movement of the elbow and had outcome scores indicative of greater disability compared to those without associated dislocation. PMID- 16365127 TI - Clinical outcome of nerve injuries associated with supracondylar fractures of the humerus in children: the experience of a specialist referral centre. AB - Between 1998 and 2002, 37 neuropathies in 32 patients with a displaced supracondylar fracture of the humerus who were referred to a nerve injury unit were identified. There were 19 boys and 13 girls with a mean age of 7.9 years (3.6 to 11.3). A retrospective review of these injuries was performed. The ulnar nerve was injured in 19, the median nerve in ten and the radial nerve in eight cases. Fourteen neuropathies were noted at the initial presentation and 23 were diagnosed after treatment of the fracture. After referral, exploration of the nerve was planned for 13 patients. Surgery was later cancelled in three because of clinical recovery. Six patients underwent neurolysis alone. Excision of neuroma and nerve grafting were performed in four. At follow-up, 26 patients had an excellent, five a good and one a fair outcome. PMID- 16365128 TI - Allograft reconstruction for bone sarcoma of the tibia in the growing child. AB - The outcome of tibial allograft reconstruction after resection of a tumour is inconsistent and has a high rate of failure. There are few reports on the use of tibial allografts in children with open growth plates. We performed 21 allograft reconstructions (16 osteoarticular, five intercalary) in 19 consecutive patients between seven and 17 years of age. Two had Ewing's sarcoma, one an adamantinoma and 16 osteosarcoma, one with multifocal disease. Five patients have died; the other 14 were free from disease at the time of follow-up. Six surviving patients (eight allograft reconstructions) continue to have good or excellent function at a mean of 59 months (14 to 132). One patient has poor function at 31 months. The other seven patients have a good or excellent function after additional procedures including exchange of the allograft and resurfacing or revision to an endoprosthesis at a mean of 101 months (43 to 198). The additional operations were performed at a mean of 47 months (20 to 84) after the first reconstruction. With the use of allograft reconstruction in growing children, joints and growth plates may be preserved, at least partially. Although our results remain inconsistent, tibial allograft reconstruction in selected patients may restore complete and durable function of the limb. PMID- 16365129 TI - Obstructive sleep apnoea syndrome in morbidly obese children with tibia vara. AB - Morbid obesity and its association with obstructive sleep apnoea syndrome have been increasingly recognised in children. Orthopaedic surgeons are often the primary medical contact for older children with tibia vara, which has long been associated with obesity, but are unfamiliar with the evaluation and treatment of sleep apnoea in children. We reviewed all children with tibia vara treated surgically at one of our institutions over a period of five years. Thirty-seven patients were identified; 18 were nine years of age or older and 13 of these (72%) had morbid obesity and a history of snoring. Eleven children were diagnosed as having sleep apnoea on polysomnography. The incidence of this syndrome in the 18 children aged nine years or older with tibia vara, was 61%. All these patients required pre-operative non-invasive positive-pressure ventilation; tonsillectomy and adenoidectomy were necessary in five (45%). No peri-operative complications related to the airway occurred. There is a high incidence of sleep apnoea in morbidly obese patients with tibia vara. These patients should be screened for snoring and, if present, should be further evaluated for sleep apnoea before corrective surgery is undertaken. PMID- 16365130 TI - Comparing ultrasound measurements of neonatal hips using the methods of Graf and Terjesen. AB - In a prospective study, 232 neonates were examined sonographically using the methods of Graf and Terjesen. In order to determine the reproducibility of the methods, 50 hips were evaluated by two skilled examiners. In an inter-observer study, five physicians and five medical students evaluated 24 images, which were evaluated on ten occasions at two-weekly intervals by one of the authors. Statistical evaluation used the Bland-Altman approach. The neonates (110 females, 122 males) were less than four days old. The mean alpha angle was 62.4 degrees and mean femoral head cover was 55.4%. According to Graf's method, 1.3% of hips were pathological, compared with 4.1% according to Terjesen. Spearman's correlation coefficient between femoral head cover and alpha angles was 0.552. The Bland-Altman approach shows greater variation for femoral head cover than for alpha, if measured by experienced examiners. The Bland-Altman approach shows almost equal reproducibility for alpha and femoral head cover in the inter observer test, but better repeatability for alpha in the intra-observer test. The Graf results relate better than Terjesen's to the well-known frequency of 1% to 2% hip dysplasia in the European population. Kappa statistics indicate a fair agreement between the two methods. Inter-observer evaluation shows an equal reproducibility of both methods, whereas intra-observer tests reveal better repeatability with Graf's method. PMID- 16365131 TI - Assessment of a self-administration protocol for extended subcutaneous thromboprophylaxis in lower limb arthroplasty. AB - The risk of venous thromboembolism in patients following arthroplasty may be reduced by continuing chemical thromboprophylaxis for up to 35 days post operatively. This prospective cohort study investigated the compliance of 40 consecutive consenting patients undergoing lower limb arthroplasty with self administration of a recommended subcutaneous chemotherapeutic agent for six weeks after surgery. Compliance was assessed by examination of the patient for signs of injection, number of syringes used, and a self-report diary at the end of the six week period. A total of 40 patients, 15 men and 25 women, were recruited. One woman was excluded because immediate post-operative complications prevented her participation. Self-administration was considered feasible in 87% of patients (95% confidence interval (CI) 76 to 98) at the time of discharge. Among this group of 34 patients, 29 (85%) were compliant (95% CI 73 to 97). Patients can learn to self-administer subcutaneous injections of thromboprophylaxis, and compliance with extended prophylaxis to six weeks is good. PMID- 16365132 TI - Maintaining standards in surgical training: how effective is the accreditation by the Specialist Advisory Committee in the United Kingdom? AB - In order to assess the efficacy of inspection and accreditation by the Specialist Advisory Committee for higher surgical training in orthopaedic surgery and trauma, seven training regions with 109 hospitals and 433 Specialist Registrars were studied over a period of two years. There were initial deficiencies in a mean of 14.8% of required standards (10.3% to 19.2%). This improved following completion of the inspection, with a mean residual deficiency in 8.9% (6.5% to 12.7%.) Overall, 84% of standards were checked, 68% of the units improved and training was withdrawn in 4%. Most units (97%) were deficient on initial assessment. Moderately good rectification was achieved but the process of follow up and collection of data require improvement. There is an imbalance between the setting of standards and their implementation. Any major revision of the process of accreditation by the new Post-graduate Medical Education and Training Board should recognise the importance of assessment of training by direct inspection on site, of the relationship between service and training, and the advantage of defining mandatory and developmental standards. PMID- 16365133 TI - Bone morphogenetic protein 7 in the treatment of congenital pseudarthrosis of the tibia. AB - We describe a 13-year-old boy with atrophic tibial pseudarthrosis associated with neurofibromatosis who had undergone nine unsuccessful operations. Eventually, union was obtained by the use of bone morphogenetic protein 7 in conjunction with intramedullary stabilisation and autologous bone graft. PMID- 16365134 TI - A comparison of the shrinkage of commercial bone cements when mixed under vacuum. AB - The outcome of a cemented hip arthroplasty is partly dependent on the type of cement which is used. The production of an interface gap between the stem and the cement mantle as a result of shrinkage of the cement, may be a factor involved. Palacos R, Palacos LV (both with gentamicin), CMW 1, CMW 2, CMW Endurance (CMWE) and Simplex were prepared under vacuum and allowed to cure overnight in similar cylinders. The next day this volume was determined by the displacement of water. Shrinkage varied between 3.82% and 7.08% with CMWE having the lowest and Palacos LV the highest. This could be a factor to consider when choosing a cement for a shape-closed stem. PMID- 16365135 TI - Histomorphometric and mechanical analysis of the hydroxyapatite-bone interface after electromagnetic stimulation: an experimental study in rabbits. AB - We investigated the effect of stimulation with a pulsed electromagnetic field on the osseointegration of hydroxyapatite in cortical bone in rabbits. Implants were inserted into femoral cortical bone and were stimulated for six hours per day for three weeks. Electromagnetic stimulation improved osseointegration of hydroxyapatite compared with animals which did not receive this treatment in terms of direct contact with the bone, the maturity of the bone and mechanical fixation. The highest values of maximum push-out force (F(max)) and ultimate shear strength (sigma(u)) were observed in the treated group and differed significantly from those of the control group at three weeks (F(max); p < 0.0001; sigma(u), p < 0.0005). PMID- 16365136 TI - Osteogenic potential of cells in vitro derived from haemarthrosis of the knee induced by injury to the anterior cruciate ligament. AB - We have investigated whether cells derived from haemarthrosis caused by injury to the anterior cruciate ligament could differentiate into the osteoblast lineage in vitro. Haemarthroses associated with anterior cruciate ligament injuries were aspirated and cultured. After treatment with beta-glycerophosphate, ascorbic acid and dexamethasone or 1,25 (OH)(2)D(3), a significant increase in the activity of alkaline phosphatase was observed. Matrix mineralisation was demonstrated after 28 days and mRNA levels in osteoblast-related genes were enhanced. Our results suggest that the haemarthrosis induced by injury to the anterior cruciate ligament contains osteoprogenitor cells and is a potential alternative source for cell-based treatment in such injury. PMID- 16365137 TI - Methicillin-resistant Staphylococcus aureus in trauma and orthopaedic practice. PMID- 16365138 TI - Factors associated with nonunion in conservatively-treated type-II fractures of the odontoid process. PMID- 16365139 TI - Obesity: a peptide YY-deficient, but not peptide YY- resistant, state. PMID- 16365140 TI - Are there more than antibodies to the thyroid-stimulating hormone receptor that meet the eye in Graves' disease? PMID- 16365141 TI - Learning hunger: conditioned anticipatory ghrelin responses in energy homeostasis. PMID- 16365142 TI - Posttranslational, reversible O-glycosylation is stimulated by high glucose and mediates plasminogen activator inhibitor-1 gene expression and Sp1 transcriptional activity in glomerular mesangial cells. AB - Metabolic flux through the hexosamine biosynthetic pathway (HBP) is increased in the presence of high glucose (HG) and potentially stimulates the expression of genes associated with the development of diabetic nephropathy. A number of synthetic processes are coupled to the HBP, including enzymatic intracellular O glycosylation (O-GlcNAcylation), the addition of single O-linked N acetylglucosamine monosaccharides to serine or threonine residues. Despite much data linking flow through the HBP and gene expression, the exact contribution of O-GlcNAcylation to HG-stimulated gene expression remains unclear. In glomerular mesangial cells, HG-stimulated plasminogen activator inhibitor-1 (PAI-1) gene expression requires the HBP and the transcription factor, Sp1. In this study, the specific role of O-GlcNAcylation in HG-induced PAI-1 expression was tested by limiting this modification with a dominant-negative O-linked N-acetylglucosamine transferase, by overexpression of neutral beta-N-acetylglucosaminidase, and by knockdown of O-linked beta-N-acetylglucosamine transferase expression by RNA interference. Decreasing O-GlcNAcylation by these means inhibited the ability of HG to increase endogenous PAI-1 mRNA and protein levels, the activity of a PAI-1 promoter-luciferase reporter gene, and Sp1 transcriptional activation. Conversely, treatment with the beta-N-acetylglucosaminidase inhibitor, O-(2 acetamido-2-deoxy-d-glucopyranosylidene)amino-N-phenylcarbamate, in the presence of normal glucose increased Sp1 O-GlcNAcylation and PAI-1 mRNA and protein levels. These findings demonstrate for the first time that among the pathways served by the HBP, O-GlcNAcylation, is obligatory for HG-induced PAI-1 gene expression and Sp1 transcriptional activation in mesangial cells. PMID- 16365144 TI - Tuberculosis in children and adults: two distinct genetic diseases. AB - Disseminated disease in children and pulmonary disease in adults constitute two major epidemiological and clinical forms of tuberculosis. Paradoxically, only a small fraction of infected individuals develop clinical tuberculosis, typically one form of the disease or the other. Mendelian and complex genetic predispositions to tuberculosis were reported recently in children and adults, respectively. Here, we argue that tuberculosis and its clinical expression largely reflect the underlying human genetic background. PMID- 16365145 TI - Functionally distinct NKT cell subsets and subtypes. AB - Natural killer T (NKT) cells are a population of autoreactive cells that mediate both protective and regulatory immune functions. NKT cells comprise several subsets of cells, but it has been unclear whether these different NKT cell subsets possess distinct functions in vivo. New studies now demonstrate that subsets of NKT cells are indeed functionally distinct and that the specific functions of these cells may be dictated in part by organ-specific mechanisms. PMID- 16365146 TI - A nonclassical non-Valpha14Jalpha18 CD1d-restricted (type II) NKT cell is sufficient for down-regulation of tumor immunosurveillance. AB - The importance of immunoregulatory T cells has become increasingly apparent. Both CD4+CD25+ T cells and CD1d-restricted NKT cells have been reported to down regulate tumor immunity in mouse tumor models. However, the relative roles of both T cell populations have rarely been clearly distinguished in the same tumor models. In addition, CD1d-restricted NKT cells have been reported to play a critical role not only in the down-regulation of tumor immunity but also in the promotion of the immunity. However, the explanation for these apparently opposite roles in different tumor models remains unclear. We show that in four mouse tumor models in which CD1d-restricted NKT cells play a role in suppression of tumor immunity, depletion of CD4+CD25+ T cells did not induce enhancement of immunosurveillance. Surprisingly, among the two subpopulations of CD1d-restricted NKT cells, Valpha14Jalpha18+ (type I) and Valpha14Jalpha18- (type II) NKT cells, type I NKT cells were not necessary for the immune suppression. These unexpected results may now resolve the paradox in the role of CD1d-restricted NKT cells in the regulation of tumor immunity, in that type II NKT cells may be sufficient for negative regulation, whereas protection has been found to be mediated by alpha galactosylceramide-responsive type I NKT cells. PMID- 16365147 TI - E47 is required for V(D)J recombinase activity in common lymphoid progenitors. AB - Common lymphoid progenitors (CLPs) are the first bone marrow precursors in which V(D)J recombinase activity is up-regulated. Here, we show that loss of the transcription factor E47 produces a reduced CLP population that lacks V(D)J recombinase activity and D-J(H) rearrangements in vivo. Apart from a profound arrest before the pro-B cell stage, other downstream lymphoid progeny of CLPs are still intact in these mice albeit at reduced numbers. In contrast to the inhibition of recombinase activity in early B lineage precursors in E47-deficient animals, loss of either E47 or its cis-acting target Erag (enhancer of rag transcription) has little effect on recombinase activity in thymic T lineage precursors. Taken together, this work defines a role for E47 in regulating lineage progression at the CLP stage in vivo and describes the first transcription factor required for lineage-specific recombinase activity. PMID- 16365148 TI - Caspase-dependent immunogenicity of doxorubicin-induced tumor cell death. AB - Systemic anticancer chemotherapy is immunosuppressive and mostly induces nonimmunogenic tumor cell death. Here, we show that even in the absence of any adjuvant, tumor cells dying in response to anthracyclins can elicit an effective antitumor immune response that suppresses the growth of inoculated tumors or leads to the regression of established neoplasia. Although both antracyclins and mitomycin C induced apoptosis with caspase activation, only anthracyclin-induced immunogenic cell death was immunogenic. Caspase inhibition by Z-VAD-fmk or transfection with the baculovirus inhibitor p35 did not inhibit doxorubicin (DX) induced cell death, yet suppressed the immunogenicity of dying tumor cells in several rodent models of neoplasia. Depletion of dendritic cells (DCs) or CD8+T cells abolished the immune response against DX-treated apoptotic tumor cells in vivo. Caspase inhibition suppressed the capacity of DX-killed cells to be phagocytosed by DCs, yet had no effect on their capacity to elicit DC maturation. Freshly excised tumors became immunogenic upon DX treatment in vitro, and intratumoral inoculation of DX could trigger the regression of established tumors in immunocompetent mice. These results delineate a procedure for the generation of cancer vaccines and the stimulation of anti-neoplastic immune responses in vivo. PMID- 16365149 TI - Carbon monoxide ameliorates chronic murine colitis through a heme oxygenase 1 dependent pathway. AB - Heme oxygenase (HO)-1 and its metabolic product carbon monoxide (CO) play regulatory roles in acute inflammatory states. In this study, we demonstrate that CO administration is effective as a therapeutic modality in mice with established chronic colitis. CO administration ameliorates chronic intestinal inflammation in a T helper (Th)1-mediated model of murine colitis, interleukin (IL)-10-deficient (IL-10(-/-)) mice. In Th1-mediated inflammation, CO abrogates the synergistic effect of interferon (IFN)-gamma on lipopolysaccharide-induced IL-12 p40 in murine macrophages and alters IFN-gamma signaling by inhibiting a member of the IFN regulatory factor (IRF) family of transcription factors, IRF-8. A specific signaling pathway, not previously identified, is delineated that involves an obligatory role for HO-1 induction in the protection afforded by CO. Moreover, CO antagonizes the inhibitory effect of IFN-gamma on HO-1 expression in macrophages. In macrophages and in Th1-mediated colitis, pharmacologic induction of HO-1 recapitulates the immunosuppressive effects of CO. In conclusion, this study begins to elucidate potential etiologic and therapeutic implications of CO and the HO-1 pathway in chronic inflammatory bowel diseases. PMID- 16365150 TI - TLR9 regulates Th1 responses and cooperates with TLR2 in mediating optimal resistance to Mycobacterium tuberculosis. AB - To investigate the role of Toll-like receptor (TLR)9 in the immune response to mycobacteria as well as its cooperation with TLR2, a receptor known to be triggered by several major mycobacterial ligands, we analyzed the resistance of TLR9(-/-) as well as TLR2/9 double knockout mice to aerosol infection with Mycobacterium tuberculosis. Infected TLR9(-/-) but not TLR2(-/-) mice displayed defective mycobacteria-induced interleukin (IL)-12p40 and interferon (IFN)-gamma responses in vivo, but in common with TLR2(-/-) animals, the TLR9(-/-) mice exhibited only minor reductions in acute resistance to low dose pathogen challenge. When compared with either of the single TLR-deficient animals, TLR2/9( /-) mice displayed markedly enhanced susceptibility to infection in association with combined defects in proinflammatory cytokine production in vitro, IFN-gamma recall responses ex vivo, and altered pulmonary pathology. Cooperation between TLR9 and TLR2 was also evident at the level of the in vitro response to live M. tuberculosis, where dendritic cells and macrophages from TLR2/9(-/-) mice exhibited a greater defect in IL-12 response than the equivalent cell populations from single TLR9-deficient animals. These findings reveal a previously unappreciated role for TLR9 in the host response to M. tuberculosis and illustrate TLR collaboration in host resistance to a major human pathogen. PMID- 16365151 TI - Fine-tuning of immune responses by SLAM-related receptors. AB - The modulation of antigen receptor signals is important for a productive immune response. The main function of the recently identified members of the signaling lymphocyte activating molecule (SLAM)-related receptors (SRR) is the fine-tuning of immune cell activation. Disruption of SRR function is the cause for severe immune disorders such as X-linked lymphoproliferative syndrome (XLP), where XLP patients carry a mutation in SLAM-associated protein (SAP) (SH2D1A), an important adaptor molecule for the signal transduction of SRR. Recent data also suggest that SRR may play a role in autoimmune diseases and the function of hematopoietic stem and progenitor cells. Here, we review the current understanding of SRR function in different immune cells. PMID- 16365152 TI - Neutrophil chemorepulsion in defined interleukin-8 gradients in vitro and in vivo. AB - We report for the first time that primary human neutrophils can undergo persistent, directionally biased movement away from a chemokine in vitro and in vivo, termed chemorepulsion or fugetaxis. Robust neutrophil chemorepulsion in microfluidic gradients of interleukin-8 (IL-8; CXC chemokine ligand 8) was dependent on the absolute concentration of chemokine, CXC chemokine receptor 2 (CXCR2), and was associated with polarization of cytoskeletal elements and signaling molecules involved in chemotaxis and leading edge formation. Like chemoattraction, chemorepulsion was pertussis toxin-sensitive and dependent on phosphoinositide-3 kinase, RhoGTPases, and associated proteins. Perturbation of neutrophil intracytoplasmic cyclic adenosine monophosphate concentrations and the activity of protein kinase C isoforms modulated directional bias and persistence of motility and could convert a chemorepellent to a chemoattractant response. Neutrophil chemorepulsion to an IL-8 ortholog was also demonstrated and quantified in a rat model of inflammation. The finding that neutrophils undergo chemorepulsion in response to continuous chemokine gradients expands the paradigm by which neutrophil migration is understood and may reveal a novel approach to our understanding of the homeostatic regulation of inflammation. PMID- 16365153 TI - Modulation of specific surface receptors and activation sensitization in primary resting CD4+ T lymphocytes by the Nef protein of HIV-1. AB - The human immunodeficiency virus type 1 (HIV-1) pathogenicity factor Nef increases viral replication in vivo. In immortalized cell lines, Nef affects the cell surface levels of multiple receptors and signal transduction pathways. Resting CD4+ T lymphocytes are important targets for HIV-1 infection in vivo-they actively transcribe and express HIV-1 genes and contribute to the local viral burden and long-lived viral reservoirs in patients undergoing antiretroviral therapy. In vitro, this primary cell type has, however, thus far been highly refractory to experimental manipulation, and the biological activities exerted by HIV-1 Nef in these cells are largely unknown. Using nucleofection for gene delivery, we find that Nef induces a drastic and moderate down-regulation of CD4 and major histocompatibility complex type 1 (MHC-I), respectively, but does not alter surface levels of other receptors, the down-modulation of which has been reported in cell line studies. In contrast, Nef markedly up-regulated cell surface levels of the MHC-II invariant chain CD74. The effect of Nef on these three surface receptors was also detected upon HIV-1 infection of activated primary CD4+ T lymphocytes. Nef expression alone was insufficient to activate resting CD4+ T lymphocytes, but Nef modestly enhanced the responsiveness of cells to exogenous T cell activation. Consistent with such a signal transduction activity, a subpopulation of Nef localized to lipid raft clusters at the plasma membrane. This study establishes the analysis of Nef functions in these primary HIV target cells. Our data support the involvement of modulation of a defined set of cell surface receptors and sensitization to activation rather than an autonomous activation function in the role of Nef in HIV-1 pathogenesis. PMID- 16365154 TI - Dendritic cells during polymicrobial sepsis rapidly mature but fail to initiate a protective Th1-type immune response. AB - Polymicrobial sepsis is associated with immunosuppression caused by the predominance of anti-inflammatory mediators and profound loss of lymphocytes through apoptosis. Dendritic cells (DC) are potent antigen-presenting cells and play a key role in T cell activation. We tested the hypothesis that DC are involved in sepsis-mediated immunosuppression in a mouse cecal ligation and puncture (CLP) model, which resembles human polymicrobial sepsis. At different time-points after CLP, DC from the spleen and peripheral lymph nodes were characterized in terms of expression of costimulatory molecules, cytokine synthesis, and subset composition. Splenic DC strongly up-regulated CD86 and CD40 but not CD80 as soon as 8 h after CLP. In contrast, lymph node DC equally increased the expression of CD86, CD40, and CD80. However, this process of maturation occurred later in the lymph nodes than in the spleen. Splenic DC from septic mice were unable to secrete interleukin (IL)-12, even upon stimulation with CpG or lipopolysaccharide+CD40 ligand, but released high levels of IL-10 in comparison to DC from control mice. Neutralization of endogenous IL-10 could not restore IL-12 secretion by DC of septic mice. In addition, the splenic CD4+CD8- and CD4-CD8+ subpopulations were lost during sepsis, and the remaining DC showed a reduced capacity for allogeneic T cell activation associated with decreased IL 2 synthesis. Thus, during sepsis, splenic DC acquire a state of aberrant responsiveness to bacterial stimuli, and two DC subtypes are selectively lost. These changes in DC behavior might contribute to impaired host response against bacteria during sepsis. PMID- 16365155 TI - Mitochondrial Ca2+ flux is a critical determinant of the Ca2+ dependence of mast cell degranulation. AB - An increase in intracellular Ca2+ ([Ca2+]i) is necessary for mast cell exocytosis, but there is controversy over the requirement for Ca2+ in the extracellular medium. Here, we demonstrate that mitochondrial function is a critical determinant of Ca2+ dependence. In the presence of extracellular Ca2+, mitochondrial metabolic inhibitors, including rotenone, antimycin A, and the protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), significantly reduced degranulation induced by immunoglobulin E (IgE) antigen or by thapsigargin, as measured by beta-hexosaminidase release. In the absence of extracellular Ca2+; however, antimycin A and FCCP, but not rotenone, enhanced, rather than reduced, degranulation to a maximum of 76% of that observed in the presence of extracellular Ca2+. This enhancement of extracellular, Ca2+ independent degranulation was concomitant with a rapid collapse of the mitochondrial transmembrane potential. Mitochondrial depolarization did not enhance degranulation induced by thapsigargin, irrespective of the presence or absence of extracellular Ca2+. IgE antigen was more effective than thapsigargin as an inducer of [Ca2+]i release, and mitochondrial depolarization augmented IgE mediated but not thapsigargin-induced Ca2+ store release and mitochondrial Ca2+ ([Ca2+]m) release. Finally, atractyloside and bongkrekic acid [an agonist and an antagonist, respectively, of the mitochondrial permeability transition pore (mPTP)], respectively, augmented and reduced IgE-mediated Ca2+ store release, [Ca2+]m release, and/or degranulation, whereas they had no effects on thapsigargin-induced Ca2+ store release. These data suggest that the mPTP is involved in the regulation of Ca2+ signaling, thereby affecting the mode of mast cell degranulation. This finding may shed light on a new role for mitochondria in the regulation of mast cell activation. PMID- 16365156 TI - Gene expression changes by amyloid beta peptide-stimulated human postmortem brain microglia identify activation of multiple inflammatory processes. AB - A central feature of the inflammatory pathology in Alzheimer's disease is activated microglia clustered around aggregated amyloid beta (Abeta) peptide containing plaques. In vitro-cultured microglia can be activated to an inflammatory state by aggregated Abeta with the induction of a range of different neurotoxic factors and provide a model system for studying microglia Abeta interactions. Gene expression responses of human postmortem brain-derived microglia to aggregated Abeta were measured using whole genome microarrays to address the hypothesis that Abeta interactions with human microglia primarily induce proinflammatory genes and not activation of genes involved in Abeta phagocytosis and removal. The results demonstrated that Abeta activation of microglia induced a large alteration in gene transcription including activation of many proinflammatory cytokines and chemokines, most notably, interleukin (IL) 1beta, IL-8, and matrix metalloproteinases (MMP), including MMP1, MMP3, MMP9, MMP10, and MMP12. All of these genes could amplify ongoing inflammation, resulting in further neuronal loss. Changes in expression of receptors associated with Abeta phagocytosis did not match the changes in proinflammatory gene expression. Time-course gene expression profiling, along with real-time polymerase chain reaction validation of expression changes, demonstrated an acute phase of gene induction for many proinflammatory genes but also chronic activation for many other potentially toxic products. These chronically activated genes included indoleamine 2,3-dioxygenase and kynureninase, which are involved in formation of the neurotoxin quinolinic acid, and S100A8, a potential proinflammatory chemokine. These studies show that activation of microglia by Abeta induces multiple genes that could be involved in inflammatory responses contributing to neurodegenerative processes. PMID- 16365157 TI - Cell surface expression of intermediate filament proteins vimentin and lamin B1 in human neutrophil spontaneous apoptosis. AB - Neutrophils represent an important source of autoantigens for antineutrophil cytoplasmic antibody associated with vasculitis. To date, two cytoskeletal proteins, vinculin and vimentin, have been reported to be expressed on the cell surfaces of activated macrophages, platelets, and apoptotic T lymphocytes. However, such cell surface expression has never been studied in human neutrophils. As we recently demonstrated that different cytoskeletal proteins were cleaved in apoptotic neutrophils, we hypothesized that some of these were expressed on the cell surface of apoptotic neutrophils. Herein, we found that among vinculin, paxillin, gelsolin, vimentin, lamin B1, alpha-tubulin, and beta tubulin, only the two intermediate filament (INFIL) proteins, vimentin and lamin B1, are expressed on the cell surface of 24-h aged neutrophils [spontaneous apoptosis (SA)]. By monitoring intracellular expression of vimentin and lamin B1 during SA, we found that these two proteins were cleaved and that such cleavage was reversed by the pan caspase inhibitor N-benzyloxy-carbonyl-V-A-D-O methylfluoromethyl ketone (z-VAD-fmk). When neutrophil apoptosis was delayed or suppressed by lipopolysaccharide or the cytokines granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage (GM)-CSF, or interleukin-4, the loss of intracellular expression of vimentin and lamin B1 was prevented. The INFIL proteins were absent from the cell surface when neutrophil apoptosis was delayed. Addition of z-VAD-fmk significantly decreased the cell surface expression of vimentin and lamin B1 during SA. This study provides the first evidence that apoptotic neutrophils express cytoskeletal proteins on their surface, opening the possibility that these cells may participate in the development of autoantibodies directed against cytoskeletal proteins, a condition frequently reported in several inflammatory diseases. PMID- 16365158 TI - Involvement of CCR5 in the passage of Th1-type cells across the blood-retina barrier in experimental autoimmune uveitis. AB - Although the recruitment of T helper cell type 1 (Th1)/Th2 cells into peripheral tissues is essential for inflammation and the host response to infection, the traffic signals that enable the distinct positioning of Th1/Th2 cells are unclear. We have determined the role of CC chemokine receptor 5 (CCR5) in this using experimental autoimmune uveitis (EAU) as a model system. In EAU, Th1-like cells are preferentially recruited into the retina across the blood-retina barrier, partly as a result of expression of the adhesion molecules P-selectin glycoprotein ligand 1 and lymphocyte function-associated antigen-1 on these cells. CD3+ T cells, infiltrating the retina, also expressed the chemokine receptor CCR5, and CCR5 ligands, macrophage-inflammatory protein-1alpha (MIP 1alpha), MIP-1beta, and regulated on activation, normal T expressed and secreted (RANTES), were strongly expressed in the retina at peak EAU. Th1-like cells, polarized in vitro, expressed high levels of CCR5. The trafficking of these CCR5+ cells was examined by tracking them after adoptive transfer in real time in vivo at an early disease stage using scanning laser ophthalmoscopy. Treatment of the cells with antibody against CCR5 prior to transfer resulted in a reduction in their infiltration into the retina. However, rolling velocity, rolling efficiency, and adherence of the cells to retinal endothelium were not reduced. CCR5 is clearly important for Th1 cell recruitment, and this study demonstrates for the first time in vivo that CCR5 may act at the level of transendothelial migration rather than at the earlier stage of rolling on the endothelium. PMID- 16365159 TI - Ligand recognition and activation of formyl peptide receptors in neutrophils. PMID- 16365160 TI - Keratin 8 overexpression promotes mouse Mallory body formation. AB - Keratins 8 and 18 (K8/18) are major constituents of Mallory bodies (MBs), which are hepatocyte cytoplasmic inclusions seen in several liver diseases. K18-null but not K8-null or heterozygous mice form MBs, which indicates that K8 is important for MB formation. Early stages in MB genesis include K8/18 hyperphosphorylation and overexpression. We used transgenic mice that overexpress K8, K18, or K8/18 to test the importance of K8 and/or K18 in MB formation. MBs were induced by feeding 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). Livers of young K8 or K8/K18 overexpressors had no histological abnormalities despite increased keratin protein and phosphorylation. In aging mice, only K8 overexpressing livers spontaneously developed small "pre-MB" aggregates. Only K8 overexpressing young mice are highly susceptible to MB formation after short-term DDC feeding. Thus, the K8 to K18 ratio, rather than K8/18 overexpression by itself, plays an essential role in MB formation. K8 overexpression is sufficient to form pre-MB and primes animals to accumulate MBs upon DDC challenge, which may help explain MB formation in human liver diseases. PMID- 16365161 TI - Tricellulin constitutes a novel barrier at tricellular contacts of epithelial cells. AB - For epithelia to function as barriers, the intercellular space must be sealed. Sealing two adjacent cells at bicellular tight junctions (bTJs) is well described with the discovery of the claudins. Yet, there are still barrier weak points at tricellular contacts, where three cells join together. In this study, we identify tricellulin, the first integral membrane protein that is concentrated at the vertically oriented TJ strands of tricellular contacts. When tricellulin expression was suppressed with RNA interference, the epithelial barrier was compromised, and tricellular contacts and bTJs were disorganized. These findings indicate the critical function of tricellulin for formation of the epithelial barrier. PMID- 16365162 TI - The mobile nucleoporin Nup2p and chromatin-bound Prp20p function in endogenous NPC-mediated transcriptional control. AB - Nuclear pore complexes (NPCs) govern macromolecular transport between the nucleus and cytoplasm and serve as key positional markers within the nucleus. Several protein components of yeast NPCs have been implicated in the epigenetic control of gene expression. Among these, Nup2p is unique as it transiently associates with NPCs and, when artificially tethered to DNA, can prevent the spread of transcriptional activation or repression between flanking genes, a function termed boundary activity. To understand this function of Nup2p, we investigated the interactions of Nup2p with other proteins and with DNA using immunopurifications coupled with mass spectrometry and microarray analyses. These data combined with functional assays of boundary activity and epigenetic variegation suggest that Nup2p and the Ran guanylyl-nucleotide exchange factor, Prp20p, interact at specific chromatin regions and enable the NPC to play an active role in chromatin organization by facilitating the transition of chromatin between activity states. PMID- 16365163 TI - Yeast phosphatidylinositol 4-kinase, Pik1, has essential roles at the Golgi and in the nucleus. AB - Phosphatidylinositol 4-kinase, Pik1, is essential for viability. GFP-Pik1 localized to cytoplasmic puncta and the nucleus. The puncta colocalized with Sec7 DsRed, a marker of trans-Golgi cisternae. Kap95 (importin-beta) was necessary for nuclear entry, but not Kap60 (importin-alpha), and exportin Msn5 was required for nuclear exit. Frq1 (frequenin orthologue) also is essential for viability and binds near the NH2 terminus of Pik1. Frq1-GFP localized to Golgi puncta, and Pik1 lacking its Frq1-binding site (or Pik1 overexpressed in frq1Delta cells) did not decorate the Golgi, but nuclear localization was unperturbed. Pik1(Delta10-192), which lacks its nuclear export sequence, displayed prominent nuclear accumulation and did not rescue inviability of pik1Delta cells. A Pik1-CCAAX chimera was excluded from the nucleus and also did not rescue inviability of pik1Delta cells. However, coexpression of Pik1(Delta10-192) and Pik1-CCAAX in pik1Delta cells restored viability. Catalytically inactive derivatives of these compartment restricted Pik1 constructs indicated that PtdIns4P must be generated both in the nucleus and at the Golgi for normal cell function. PMID- 16365164 TI - Transition from hemifusion to pore opening is rate limiting for vacuole membrane fusion. AB - Fusion pore opening and expansion are considered the most energy-demanding steps in viral fusion. Whether this also applies to soluble N-ethyl-maleimide sensitive fusion protein attachment protein receptor (SNARE)- and Rab-dependent fusion events has been unknown. We have addressed the problem by characterizing the effects of lysophosphatidylcholine (LPC) and other late-stage inhibitors on lipid mixing and pore opening during vacuole fusion. LPC inhibits fusion by inducing positive curvature in the bilayer and changing its biophysical properties. The LPC block reversibly prevented formation of the hemifusion intermediate that allows lipid, but not content, mixing. Transition from hemifusion to pore opening was sensitive to guanosine-5'-(gamma-thio)triphosphate. It required the vacuolar adenosine triphosphatase V0 sector and coincided with its transformation. Pore opening was rate limiting for the reaction. As with viral fusion, opening the fusion pore may be the most energy-demanding step for intracellular, SNARE dependent fusion reactions, suggesting that fundamental aspects of lipid mixing and pore opening are related for both systems. PMID- 16365165 TI - Influences on neural lineage and mode of division in the zebrafish retina in vivo. AB - Cell determination in the retina has been under intense investigation since the discovery that retinal progenitors generate clones of apparently random composition (Price, J., D. Turner, and C. Cepko. 1987. Proc. Natl. Acad. Sci. USA. 84:156-160; Holt, C.E., T.W. Bertsch, H.M. Ellis, and W.A. Harris. 1988. Neuron. 1:15-26; Wetts, R., and S.E. Fraser. 1988. Science. 239:1142-1145). Examination of fixed tissue, however, sheds little light on lineage patterns or on the relationship between the orientation of division and cell fate. In this study, three-dimensional time-lapse analyses were used to trace lineages of retinal progenitors expressing green fluorescent protein under the control of the ath5 promoter. Surprisingly, these cells divide just once along the circumferential axis to produce two postmitotic daughters, one of which becomes a retinal ganglion cell (RGC). Interestingly, when these same progenitors are transplanted into a mutant environment lacking RGCs, they often divide along the central-peripheral axis and produce two RGCs. This study provides the first insight into reproducible lineage patterns of retinal progenitors in vivo and the first evidence that environmental signals influence the orientation of cell division and the lineage of neural progenitors. PMID- 16365166 TI - Expression of mutant huntingtin in glial cells contributes to neuronal excitotoxicity. AB - Huntington disease (HD) is characterized by the preferential loss of striatal medium-sized spiny neurons (MSNs) in the brain. Because MSNs receive abundant glutamatergic input, their vulnerability to excitotoxicity may be largely influenced by the capacity of glial cells to remove extracellular glutamate. However, little is known about the role of glia in HD neuropathology. Here, we report that mutant huntingtin accumulates in glial nuclei in HD brains and decreases the expression of glutamate transporters. As a result, mutant huntingtin (htt) reduces glutamate uptake in cultured astrocytes and HD mouse brains. In a neuron-glia coculture system, wild-type glial cells protected neurons against mutant htt-mediated neurotoxicity, whereas glial cells expressing mutant htt increased neuronal vulnerability. Mutant htt in cultured astrocytes decreased their protection of neurons against glutamate excitotoxicity. These findings suggest that decreased glutamate uptake caused by glial mutant htt may critically contribute to neuronal excitotoxicity in HD. PMID- 16365167 TI - Raf-1 sets the threshold of Fas sensitivity by modulating Rok-alpha signaling. AB - Ablation of the Raf-1 protein causes fetal liver apoptosis, embryonic lethality, and selective hypersensitivity to Fas-induced cell death. Furthermore, Raf-1 deficient cells show defective migration as a result of the deregulation of the Rho effector kinase Rok-alpha. In this study, we show that the kinase-independent modulation of Rok-alpha signaling is also the basis of the antiapoptotic function of Raf-1. Fas activation stimulates the formation of Raf-1-Rok-alpha complexes, and Rok-alpha signaling is up-regulated in Raf-1-deficient cells. This leads to increased clustering and membrane expression of Fas, which is rescued both by kinase-dead Raf-1 and by interfering with Rok-alpha or its substrate ezrin. Increased Fas clustering and membrane expression are also evident in the livers of Raf-1-deficient embryos, and genetically reducing Fas expression counteracts fetal liver apoptosis, embryonic lethality, and the apoptotic defects of embryonic fibroblasts. Thus, Raf-1 has an essential function in regulating Fas expression and setting the threshold of Fas sensitivity during embryonic life. PMID- 16365168 TI - Distinct roles of Akt1 and Akt2 in regulating cell migration and epithelial mesenchymal transition. AB - The Akt family of kinases are activated by growth factors and regulate pleiotropic cellular activities. In this study, we provide evidence for isoform specific positive and negative roles for Akt1 and -2 in regulating growth factor stimulated phenotypes in breast epithelial cells. Insulin-like growth factor-I receptor (IGF-IR) hyperstimulation induced hyperproliferation and antiapoptotic activities that were reversed by Akt2 down-regulation. In contrast, Akt1 down regulation in IGF-IR-stimulated cells promoted dramatic neomorphic effects characteristic of an epithelial-mesenchymal transition (EMT) and enhanced cell migration induced by IGF-I or EGF stimulation. The phenotypic effects of Akt1 down-regulation were accompanied by enhanced extracellular signal-related kinase (ERK) activation, which contributed to the induction of migration and EMT. Interestingly, down-regulation of Akt2 suppressed the EMT-like morphological conversion induced by Akt1 down-regulation in IGF-IR-overexpressing cells and inhibited migration in EGF-stimulated cells. These results highlight the distinct functions of Akt isoforms in regulating growth factor-stimulated EMT and cell migration, as well as the importance of Akt1 in cross-regulating the ERK signaling pathway. PMID- 16365169 TI - Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. AB - The intermediate filament (IF)-binding protein desmoplakin (DP) is essential for desmosome function and tissue integrity, but its role in junction assembly is poorly understood. Using time-lapse imaging, we show that cell-cell contact triggers three temporally overlapping phases of DP-GFP dynamics: (1) the de novo appearance of punctate fluorescence at new contact zones after as little as 3 min; (2) the coalescence of DP and the armadillo protein plakophilin 2 into discrete cytoplasmic particles after as little as 15 min; and (3) the cytochalasin-sensitive translocation of cytoplasmic particles to maturing borders, with kinetics ranging from 0.002 to 0.04 microm/s. DP mutants that abrogate or enhance association with IFs exhibit delayed incorporation into junctions, altering particle trajectory or increasing particle pause times, respectively. Our data are consistent with the idea that DP assembles into nascent junctions from both diffusible and particulate pools in a temporally overlapping series of events triggered by cell-cell contact and regulated by actin and DP-IF interactions. PMID- 16365171 TI - JCO and the Public Trust. PMID- 16365170 TI - Alpha4beta1-dependent adhesion strengthening under mechanical strain is regulated by paxillin association with the alpha4-cytoplasmic domain. AB - The capacity of integrins to mediate adhesiveness is modulated by their cytoplasmic associations. In this study, we describe a novel mechanism by which alpha4-integrin adhesiveness is regulated by the cytoskeletal adaptor paxillin. A mutation of the alpha4 tail that disrupts paxillin binding, alpha4(Y991A), reduced talin association to the alpha4beta1 heterodimer, impaired integrin anchorage to the cytoskeleton, and suppressed alpha4beta1-dependent capture and adhesion strengthening of Jurkat T cells to VCAM-1 under shear stress. The mutant retained intrinsic avidity to soluble or bead-immobilized VCAM-1, supported normal cell spreading at short-lived contacts, had normal alpha4-microvillar distribution, and responded to inside-out signals. This is the first demonstration that cytoskeletal anchorage of an integrin enhances the mechanical stability of its adhesive bonds under strain and, thereby, promotes its ability to mediate leukocyte adhesion under physiological shear stress conditions. PMID- 16365172 TI - Revisions of and clarifications to the ASCO conflict of interest policy. PMID- 16365173 TI - American Society Of Clinical Oncology: revised conflict of interest policy. PMID- 16365174 TI - Thalidomide and dexamethasone: a new standard of care for initial therapy in multiple myeloma. PMID- 16365175 TI - Intensive surveillance after stage II or III colorectal cancer: is it worth it? PMID- 16365176 TI - Lung cancer and race: equal treatment yields equal outcome among equal patients, but there is no equal treatment. PMID- 16365177 TI - When does the presence of the target predict response to the targeted agent? PMID- 16365178 TI - Phase III clinical trial of thalidomide plus dexamethasone compared with dexamethasone alone in newly diagnosed multiple myeloma: a clinical trial coordinated by the Eastern Cooperative Oncology Group. AB - PURPOSE: To determine if thalidomide plus dexamethasone yields superior response rates compared with dexamethasone alone as induction therapy for newly diagnosed multiple myeloma. PATIENTS AND METHODS: Patients were randomly assigned to receive thalidomide plus dexamethasone or dexamethasone alone. Patients in arm A received thalidomide 200 mg orally for 4 weeks; dexamethasone was administered at a dose of 40 mg orally on days 1 to 4, 9 to 12, and 17 to 20. Cycles were repeated every 4 weeks. Patients in arm B received dexamethasone alone at the same schedule as in arm A. RESULTS: Two hundred seven patients were enrolled: 103 were randomly assigned to thalidomide plus dexamethasone and 104 were randomly assigned to dexamethasone alone; eight patients were ineligible. The response rate with thalidomide plus dexamethasone was significantly higher than with dexamethasone alone (63% v 41%, respectively; P = .0017). The response rate allowing for use of serum monoclonal protein levels when a measurable urine monoclonal protein was unavailable at follow-up was 72% v 50%, respectively. The incidence rates of grade 3 or higher deep vein thrombosis (DVT), rash, bradycardia, neuropathy, and any grade 4 to 5 toxicity in the first 4 months were significantly higher with thalidomide plus dexamethasone compared with dexamethasone alone (45% v 21%, respectively; P < .001). DVT was more frequent in arm A than in arm B (17% v 3%); grade 3 or higher peripheral neuropathy was also more frequent (7% v 4%, respectively). CONCLUSION: Thalidomide plus dexamethasone demonstrates significantly superior response rates in newly diagnosed myeloma compared with dexamethasone alone. However, this must be balanced against the greater toxicity seen with the combination. PMID- 16365179 TI - Tumor necrosis factor-alpha-induced protein 3 as a putative regulator of nuclear factor-kappaB-mediated resistance to O6-alkylating agents in human glioblastomas. AB - PURPOSE: Pre-existing and acquired drug resistance are major obstacles to the successful treatment of glioblastomas. METHODS: We used an integrated resistance model and genomics tools to globally explore molecular factors and cellular pathways mediating resistance to O6-alkylating agents in glioblastoma cells. RESULTS: We identified a transcriptomic signature that predicts a common in vitro and in vivo resistance phenotype to these agents, a proportion of which is imprinted recurrently by gene dosage changes in the resistant glioblastoma genome. This signature was highly enriched for genes with functions in cell death, compromise, and survival. Modularity was a predominant organizational principle of the signature, with functions being carried out by groups of interacting molecules in overlapping networks. A highly significant network was built around nuclear factor-kappaB (NF-kappaB), which included the persistent alterations of various NF-kappaB pathway elements. Tumor necrosis factor-alpha induced protein 3 (TNFAIP3) was identified as a new regulatory component of a putative cytoplasmic signaling cascade that mediates NF-kappaB activation in response to DNA damage caused by O6-alkylating agents. Expression of the corresponding zinc finger protein A20 closely mirrored the expression of the TNFAIP3 transcript, and was inversely related to NF-kappaB activation status in the resistant cells. A prediction model based on the resistance signature enabled the subclassification of an independent, validation cohort of 31 glioblastomas into two outcome groups (P = .037) and revealed TNFAIP3 as part of an optimized four-gene predictor associated significantly with patient survival (P = .022). CONCLUSION: Our results offer strong evidence for TNFAIP3 as a key regulator of the cytoplasmic signaling to activate NF-kappaB en route to O6-alkylating agent resistance in glioblastoma cells. This pathway may be an attractive target for therapeutic modulation of glioblastomas. PMID- 16365180 TI - The effect of race on invasive staging and surgery in non-small-cell lung cancer. AB - PURPOSE: Black patients with early-stage non-small-cell lung cancer (NSCLC) have worse overall survival than white patients. Decreased likelihood of resection has been implicated. To isolate the effect of decision making from access to care, we used receipt of surgical staging as a proxy for access and willingness to undergo invasive procedures, and examined treatments and outcomes by race. PATIENTS AND METHODS: We examined registry and claims data of Medicare-eligible patients with nonmetastatic NSCLC in areas monitored by the Surveillance, Epidemiology, and End Results program from 1991 to 2001. Patients who obtained invasive staging, defined as bronchoscopy, mediastinoscopy, or thoracoscopy, were included. Logistic regression and Cox modeling calculated the odds of having staging and surgery, and survival outcomes. RESULTS: A total of 14,224 patients underwent staging, and 6,972 had surgery for lung cancer. Black patients were less likely to undergo staging (odds ratio [OR] = 0.75; 95% CI, 0.67 to 0.83), and once staged, were still less likely to have surgery than whites (OR = 0.55; 95% CI, 0.47 to 0.64). Survival for blacks and whites was equivalent after resection (hazard ratio = 1.02; P = .06). Staged black patients were less likely to receive a recommendation for surgery when it was not clearly contraindicated (67.0% v 71.4%; P < .05), and were more likely to decline surgery (3.4% v 2.0%; P < .05). CONCLUSION: Black patients obtain surgery for lung cancer less often than whites, even after access to care has been demonstrated. They are more likely not to have surgery recommended, and more likely to refuse surgery. Additional research should focus on the physician-patient encounter as a potential source of racial disparities. PMID- 16365181 TI - Similar outcomes between African American and non-African American patients with extensive-stage small-cell lung carcinoma: report from the Cancer and Leukemia Group B. AB - PURPOSE: Among patients with small-cell lung carcinoma, African Americans have lower survival rates than non-African Americans. We investigated whether the disparity in survival would persist when patients were treated with similar therapies (ie, phase II/III Cancer and Leukemia Group B [CALGB] trials). PATIENTS AND METHODS: We assessed 995 patients (928 non-African American and 67 African American) receiving chemotherapy in CALGB studies for extensive-stage small-cell lung cancer (ES-SCLC). Clinical and demographic characteristics, treatment received, and survival data were obtained from the CALGB database. The Cox proportional hazards model was used to assess the effect of race/ethnicity on survival after adjustment for other known prognostic factors. All statistical tests were two sided. RESULTS: The unadjusted survival distribution of African American patients was not significantly different from that of non-African American patients; median survival was 11.5 months (95% CI, 9.4 to 13.4 months) for African American patients versus 9.9 months (95% CI, 9.6 to 10.3 months) for non-African American patients. Multivariable adjustment for the effect of treatment arm, histology, and metastatic site at presentation did not alter the outcome for African American patients. Survival was similar even though African American patients were more likely to have a poorer performance status, present with significant weight loss, and be Medicaid recipients (20% v 6%), which is an indicator of lower socioeconomic status. CONCLUSION: African American patients tended to present with prognostic features associated with a worse survival. However, when offered equivalent therapy, the outcome for African American patients was the same as that observed for non-African American patients. PMID- 16365182 TI - Postoperative surveillance in patients with colorectal cancer who have undergone curative resection: a prospective, multicenter, randomized, controlled trial. AB - PURPOSE: Although systematic postoperative surveillance of patients with colorectal cancer has been demonstrated to improve survival, it remains unknown whether a more intensive strategy provides any significant advantage. This prospective, multicenter, randomized, controlled trial was aimed at comparing the efficacy of two different surveillance strategies in terms of both survival and recurrence resectability. PATIENTS AND METHODS: Patients with stage II or III colorectal cancer were allocated randomly to either a simple surveillance strategy including clinical evaluation and serum carcinoembryonic antigen monitoring, or an intensive strategy in which abdominal computed tomography or ultrasonography, chest radiograph, and colonoscopy were added. RESULTS: A total of 259 patients were included: 132 were observed according to the simple strategy and 127 were observed according to the intensive strategy. Both groups were similar with respect to baseline characteristics and rate and type of tumor recurrence. After a median follow-up of 48 months, there was no difference in the probability of overall survival in the whole series (hazard ratio [HR] = 0.87; 95% CI, 0.49 to 1.54; P = .62). However, the intensive strategy was associated with higher overall survival in patients with stage II tumors (HR = 0.34; 95% CI, 0.12 to 0.98; P = .045) and in those with rectal lesions (HR = 0.09; 95% CI, 0.01 to 0.81; P = .03), mainly due to higher rate of resectability for recurrent tumors. Colonoscopy was responsible for the detection of the highest proportion (44%) of resectable tumor recurrence in the intensive arm. CONCLUSION: A more intensive surveillance strategy improves the prognosis of patients with stage II colorectal cancer or those with rectal tumors. Inclusion of regular performance of colonoscopy seems justified up to the fifth year of follow-up, at least. PMID- 16365183 TI - Impact of vascular endothelial growth factor-A expression, thrombospondin-2 expression, and microvessel density on the treatment effect of bevacizumab in metastatic colorectal cancer. AB - PURPOSE: Bevacizumab is a monoclonal antibody to vascular endothelial growth factor-A (VEGF). In the pivotal trial in metastatic colorectal cancer (mCRC), addition of bevacizumab to first-line irinotecan, fluorouracil, and leucovorin (IFL) significantly prolonged median survival. The aim of these retrospective subset analyses was to evaluate VEGF, thrombospondin-2 (THBS-2), and microvessel density (MVD) as prognostic factors and/or predictors of benefit from bevacizumab. PATIENTS AND METHODS: In the pivotal trial, 813 patients with untreated mCRC were randomly assigned to receive IFL plus bevacizumab or placebo. Of 312 tissue samples collected (285 primaries, 27 metastases), outcome data were available for 278 (153 bevacizumab, 125 placebo). Epithelial and stromal VEGF expression were assessed by in situ hybridization (ISH) and immunohistochemistry on tissue microarrays and whole sections. Stromal THBS-2 expression was examined by ISH on tissue microarrays. MVD was quantified by Chalkley count. Overall survival was associated with these variables in retrospective subset analyses. RESULTS: In all subgroups, estimated hazard ratios (HRs) for risk of death were < 1 for bevacizumab-treated patients regardless of the level of VEGF or THBS-2 expression or MVD. Patients with a high THBS-2 score showed a nonsignificant improvement in survival following bevacizumab treatment (HR = 0.11; 95% CI, 0.02 to 0.51) compared to patients with a low score (HR = 0.65; 95% CI, 0.41 to 1.02); interaction analysis P = .22. VEGF or THBS-2 expression and MVD were not significant prognostic factors. CONCLUSION: These exploratory analyses suggest that in patients with mCRC addition of bevacizumab to IFL improves survival regardless of the level of VEGF or THBS-2 expression, or MVD. PMID- 16365184 TI - A role for p38 mitogen-activated protein kinase and c-myc in endothelin-dependent rat aortic smooth muscle cell proliferation. AB - We have demonstrated recently that endothelin (ET) stimulates rat aortic smooth muscle cell proliferation through an extracellular signal-regulated kinase (ERK) dependent mechanism. Approximately 70% of ET-dependent [3H]-thymidine incorporation in these cells signals through this system. In the present study, we show that the residual mitogenic activity requires an intact p38 mitogen activated protein kinase (p38 MAPK) system and increased c-myc gene expression. ET increased [3H]-thymidine incorporation in rat aortic smooth muscle cells approximately 5-fold. p38 MAPK inhibition with SB203580 or ERK/ERK kinase inhibition with PD98059 each effected approximately 70% inhibition in ET dependent DNA synthesis, whereas the combination led to nearly complete blockade of the ET effect. ET also increased c-myc RNA levels and c-Myc protein levels in these cells. The increment in c-Myc expression was blocked by SB203580 but not by PD98059. Use of antisense oligonucleotides directed against the translation start site of the c-myc transcript, but not scrambled oligonucleotide sequence, resulted in approximately 60% decrease in ET-dependent [3H]-thymidine incorporation. The combination of antisense c-myc and PD98059 resulted in near complete inhibition of ET-dependent DNA synthesis. Both ET and c-Myc increased expression and promoter activity of E2F, a transcription factor that has been linked to enhanced cell cycle activity. The ET-dependent increment in E2F promoter activity was suppressed after treatment with SB203580 or antisense c-myc but not by PD98059 or a scrambled oligonucleotide sequence. Collectively, these findings demonstrate that ET uses 2 complementary signal transduction cascades (ERK and p38 MAPK) to control proliferative activity of vascular smooth muscle cells. PMID- 16365187 TI - A novel mechanism contributing to development of Dahl salt-sensitive hypertension: role of the transient receptor potential vanilloid type 1. AB - To determine the role of the transient receptor potential vanilloid type 1 (TRPV1) channels in development of hypertension in Dahl salt-sensitive (DS) rats fed a high-salt diet (HS), male DS and Dahl salt-resistant (DR) rats were maintained on a low-salt diet (LS) or HS for 3 weeks. HS significantly increased systolic blood pressure in DS+HS rats compared with DS+LS, DR+HS, and DR+LS rats. Intravenous bolus injection of capsazepine (3 mg/kg), a selective TRPV1 antagonist, significantly increased mean arterial pressure in conscious DR+HS rats compared with DR+LS, DS+/-HS, and DS+/-LS rats. In contrast, capsaicin (10 or 30 microg/kg), a selective TRPV1 agonist, dose-dependently decreased mean arterial pressure in all of the groups with the most profound magnitude in DR+HS rats compared with the other 3 groups. TRPV1 expression in mesenteric resistance arteries and the renal cortex and medulla, calcitonin gene-related peptide levels in dorsal root ganglia, and calcitonin gene-related peptide-positive sensory nerve density in mesenteric resistance arteries were significantly decreased in DS+HS rats compared with DS+LS, DR+HS, and DR+LS rats. Taken together, our data indicate that the TRPV1 receptor is activated and its expression upregulated during HS intake in DR rats, which acts to prevent salt-induced increases in blood pressure. In contrast, TRPV1 expression and function are impaired in DS rats, which renders DS rats sensitive to salt load in terms of blood pressure regulation. PMID- 16365188 TI - Pancreatic polypeptide-fold peptide receptors and angiotensin II-induced renal vasoconstriction. AB - The Gi pathway augments renal vasoconstriction induced by angiotensin II in spontaneously hypertensive but not normotensive Wistar-Kyoto rats. Because the Gi coupled pancreatic polypeptide (PP)-fold peptide receptors Y1 and Y2 are expressed in kidneys and are activated by endogenous PP-fold peptides, we tested the hypothesis that these receptors regulate angiotensin II-induced renal vasoconstriction in kidneys from hypertensive but not normotensive rats. A selective Y1-receptor agonist [(Leu31,Pro34)-neuropeptide Y; 6 to 10 nmol/L] greatly potentiated angiotensin II-induced changes in perfusion pressure in isolated, perfused kidneys from hypertensive but not normotensive rats. A selective Y2-receptor agonist (peptide YY(3-36); 6 nM) only slightly potentiated angiotensin II-induced renal vasoconstriction and only in kidneys from hypertensive rats. Neither the Y1-receptor nor the Y2-receptor agonist increased basal perfusion pressure. BIBP3226 (1 micromol/L, highly selective Y1-receptor antagonist) and BIIE0246 (1 micromol/L, highly selective Y2-receptor antagonist) completely abolished potentiation by (Leu31,Pro34)-neuropeptide Y and peptide YY(3-36), respectively. Y1-receptor and Y2-receptor mRNA and protein levels were expressed in renal microvessels and whole kidneys, but the abundance was similar in kidneys from hypertensive and normotensive rats. Both Y1-receptor-induced and Y2-receptor-induced potentiation of angiotensin II-mediated renal vasoconstriction was completely abolished by pretreatment with pertussis toxin (30 microg/kg IV, blocks Gi proteins). These data indicate that, in kidneys from genetically hypertensive but not normotensive rats, Y1-receptor activation markedly enhances angiotensin II-mediated renal vasoconstriction by a mechanism involving Gi. Although Y2 receptors can also potentiate angiotensin II-mediated renal vasoconstriction via Gi, the effect is modest compared with Y1 receptors. These findings may have important implications for the etiology of genetic hypertension. PMID- 16365189 TI - Sodium bicarbonate cotransporter polymorphisms are associated with baseline and 10-year follow-up blood pressures. AB - The NaHCO3 cotransporter gene (SLC4A5) on chromosome 2 encodes a protein that transports sodium and bicarbonate across the cell membrane and regulates cellular pH. The National Heart, Lung, and Blood Institute Family Blood Pressure Program found linkage of blood pressure-related traits to the chromosomal region containing SLC4A5 and phenotype associations with single nucleotide polymorphisms (SNPs) in this gene. However, the results were inconsistent over various phenotypes and SNPs. Nevertheless, the evidence was strong enough to propose this gene as a blood pressure-related gene. To extend these findings, SLC4A5 SNPs were genotyped in an independent set of 96 Utah pedigrees of 1040 adult subjects at baseline, 760 of whom were followed longitudinally for 10 years. After adjusting for age, gender, body mass index, and polygenic correlations within pedigrees, SNP hcv1137534 was significantly associated with both systolic blood pressure and diastolic blood pressure (DBP) at baseline (unadjusted P=0.009 and P=0.043; respectively) and at 10-year follow-up (P=0.008 and P=0.007; respectively). In secondary tests of association of baseline-stressed blood pressure, hcv1137534 was borderline or significantly associated with DBP change during an isometric handgrip test (P=0.054), DBP change from supine to standing (P=0.020), and DBP change after a 50 degrees tilt (P=0.034). There was no evidence for compensation of abnormal SLC4A5 sodium transport by genotype-specific differences in sodium lithium countertransport, lithium-potassium cotransport, altered plasma sodium, chloride, or CO2 levels. Therefore, in these Utah pedigrees, the SLC4A5 gene was significantly associated with blood pressure and persisted after 10 years of follow-up. These results additionally confirm the involvement of SLC4A5 with blood pressure control, although the mechanism is still unclear. PMID- 16365190 TI - Regulation of peroxisome proliferator-activated receptor gamma activity by losartan metabolites. AB - Two active metabolites of the angiotensin type 1 (AT1) receptor blocker losartan have been described previously, EXP3174 and EXP3179. Whereas EXP3174 is the main antihypertensive AT1 receptor-blocking metabolite, the role of EXP3179 is widely unknown. Recently, a subgroup of AT1 receptor blockers has been identified as ligands for the peroxisome proliferator-activated receptor gamma (PPAR-gamma). Here we characterize the PPAR-gamma-activating properties of the 2 active losartan metabolites. PPAR-gamma activity was measured with a chimeric Gal4-DNA binding domain-hPPARgamma-ligand-binding domain (LBD) fusion protein on a Gal4 dependent luciferase reporter system. EXP3179 prominently induced the activation of the PPAR-gamma-LBD reaching a maximum at 100 micromol/L with a 7.1+/-1-fold induction (P<0.05 versus vehicle-treated cells). Maximum PPAR-gamma-LBD activation by EXP3179 reached 51% of the maximum response induced by the full PPAR-gamma agonist pioglitazone, identifying EXP3179 as a partial PPAR-gamma agonist. EXP3174 did not induce PPAR-gamma-LBD activation. EC50 values were calculated for PPAR-gamma-LBD activity (pioglitazone EC50: 0.88 micromol/L; EXP3179 EC50: 17.1 micromol/L; losartan EC50: >50 micromol/L). Consistent with the activation of PPAR-gamma, EXP3179 potently induced 3T3-L1 adipocyte differentiation, a typical PPAR-gamma-dependent cell function, and markedly stimulated PPAR-gamma target gene expression. EXP3174 failed to regulate differentiation or PPAR-gamma target gene expression. The present study characterizes the active losartan metabolite EXP3179 as a partial PPAR-gamma agonist. PPAR-gamma activation by EXP3179 may help us to understand the beneficial metabolic effects of losartan observed in clinical trials. PMID- 16365191 TI - Mineralocorticoid receptor activation causes cerebral vessel remodeling and exacerbates the damage caused by cerebral ischemia. AB - Mineralocorticoid receptor antagonists protect against ischemic cerebrovascular disease; this appears to be caused by changes in cerebral vessel structure that would promote blood flow. Therefore, we hypothesized that mineralocorticoid receptor activation with deoxycorticosterone acetate would cause deleterious remodeling of the cerebral vasculature and exacerbate the damage caused by cerebral ischemia. Six-week-old male Wistar rats were treated with deoxycorticosterone acetate (200 mg/kg) for 6 weeks. At 12 weeks of age, the deoxycorticosterone acetate-treated rats had elevated systolic blood pressure compared with age-matched controls (157+/-5.9 versus 124+/-3.1 mm Hg deoxycorticosterone acetate versus control; P<0.05). The area of ischemic damage resulting from middle cerebral artery occlusion was greater in the deoxycorticosterone acetate-treated rats than control (63.5+/-3.72 versus 46.6+/ 5.52% of the hemisphere infarcted, deoxycorticosterone acetate versus control; P<0.05). Middle cerebral artery structure was assessed using a pressurized arteriograph under calcium-free conditions. Over a range of intralumenal pressures, the lumen and ODs of the middle cerebral arteries were smaller in the deoxycorticosterone acetate-treated rats than the control rats (P<0.05). There was also an increase in the wall thickness and wall:lumen ratio in the vessels from deoxycorticosterone acetate-treated rats (P<0.05). The vessels from the deoxycorticosterone acetate-treated rats were stiffer than those from control rats as evidenced by a leftward shift in the stress/strain curve. These novel data suggest that mineralocorticoid receptor activation without salt loading and nephrectomy is sufficient to elicit deleterious effects on the cerebral vasculature that lead to inward hypertrophic remodeling and an increase in the ischemic damage in the event of a stroke. PMID- 16365192 TI - Angiotensin-converting enzyme 2 and angiotensin-(1-7): an evolving story in cardiovascular regulation. AB - This lecture summarizes the chronology and rationale that led to the discovery of angiotensin-(1-7) as a hormone that, in its own right, opposes the vasoconstrictor and proliferative actions of angiotensin II. The work discussed here additionally analyzes the newest findings on angiotensin-converting enzyme 2, the angiotensin-converting enzyme homologue that efficiently hydrolyzes angiotensin II into angiotensin-(1-7). Both components of this system may significantly influence our future perspective of the role of the renin angiotensin system, not just in terms of its role in the regulation of cardiovascular and renal function but, moreover, as regulators of a vast array of disease processes in which inflammation and immune mechanisms play a role. PMID- 16365193 TI - Views of the renin-angiotensin system: brilling, mimsy, and slithy tove. AB - The renin-angiotensin system plays a role in many physiological systems, as proven by the phenotype of angiotensin-converting enzyme (ACE) knockout mice. We have used homologous recombination to create novel lines of mice with limited and unusual expression patterns of ACE. These mice show that, as long as an animal can regulate renin, they can tolerate both unusual patterns and reduced expression of ACE. We have also created mice in which one of the two ACE catalytic sites is nonfunctional. These new lines of mice give great insight into the function of the renin-angiotensin system in blood pressure control, response to stress, hematopoiesis, and reproduction. PMID- 16365194 TI - Distribution of coronary artery calcium by race, gender, and age: results from the Multi-Ethnic Study of Atherosclerosis (MESA). AB - BACKGROUND: Coronary artery calcium (CAC) has been demonstrated to be associated with the risk of coronary heart disease. The Multi-Ethnic Study of Atherosclerosis (MESA) provides a unique opportunity to examine the distribution of CAC on the basis of age, gender, and race/ethnicity in a cohort free of clinical cardiovascular disease and treated diabetes. METHODS AND RESULTS: MESA is a prospective cohort study designed to investigate subclinical cardiovascular disease in a multiethnic cohort free of clinical cardiovascular disease. The percentiles of the CAC distribution were estimated with nonparametric techniques. Treated diabetics were excluded from analysis. There were 6110 included in the analysis, with 53% female and an average age of 62 years. Men had greater calcium levels than women, and calcium amount and prevalence were steadily higher with increasing age. There were significant differences in calcium by race, and these associations differed across age and gender. For women, whites had the highest percentiles and Hispanics generally had the lowest; in the oldest age group, however, Chinese women had the lowest values. Overall, Chinese and black women were intermediate, with their order dependent on age. For men, whites consistently had the highest percentiles, and Hispanics had the second highest. Blacks were lowest at the younger ages, and Chinese were lowest at the older ages. At the MESA public website (http://www.mesa-nhlbi.org), an interactive form allows one to enter an age, gender, race/ethnicity, and CAC score to obtain a corresponding estimated percentile. CONCLUSIONS: The information provided here can be used to examine whether a patient has a high CAC score relative to others with the same age, gender, and race/ethnicity who do not have clinical cardiovascular disease or treated diabetes. PMID- 16365195 TI - Electrocardiographic strain pattern and prediction of new-onset congestive heart failure in hypertensive patients: the Losartan Intervention for Endpoint Reduction in Hypertension (LIFE) study. AB - BACKGROUND: The ECG strain pattern of ST depression and T-wave inversion is strongly associated with left ventricular hypertrophy (LVH) independently of coronary heart disease and with an increased risk of cardiovascular morbidity and mortality in hypertensive patients. However, whether ECG strain is an independent predictor of new-onset congestive heart failure (CHF) in the setting of aggressive antihypertensive therapy in unclear. METHODS AND RESULTS: The relationship of ECG strain at study baseline to the development of CHF was examined in 8696 patients with no history of CHF who were enrolled in the Losartan Intervention for Endpoint Reduction in Hypertension (LIFE) study. All patients had ECG LVH by Cornell product and/or Sokolow-Lyon voltage criteria on a screening ECG, were treated in a blinded manner with atenolol- or losartan-based regimens, and were followed up for a mean of 4.7+/-1.1 years. Strain was defined as a downsloping convex ST segment with inverted asymmetrical T-wave opposite the QRS axis in lead V5 or V6. ECG strain was present in 923 patients (10.6%), and new-onset CHF occurred in 265 patients (3.0%), 26 of whom had a CHF-related death. Compared with patients who did not develop CHF, hypertensive patients who developed CHF were older; were more likely to be black, current smokers, and diabetic; were more like to have a history of myocardial infarction, ischemic heart disease, stroke, or peripheral vascular disease; and had greater baseline severity of LVH by Cornell product and Sokolow-Lyon voltage, higher baseline body mass indexes, higher serum glucose levels and albuminuria, similar baseline systolic and diastolic pressures, and reductions in diastolic pressure with treatment but greater reductions in systolic pressure. In univariate Cox analyses, ECG strain was a significant predictor of new-onset CHF (hazard ratio [HR], 3.27; 95% CI, 2.49 to 4.29) and CHF mortality (HR, 4.74; 95% CI, 2.11 to 10.64). In Cox multivariable analyses adjusting for baseline differences between patients with and without new-onset CHF, in-treatment differences in systolic and diastolic pressures, Sokolow-Lyon voltage, and Cornell product, and the impact of treatment with losartan versus atenolol on outcomes, ECG strain remained a significant predictor of incident CHF (HR, 1.80; 95% CI, 1.30 to 2.48) and CHF related death (HR, 2.78; 95% CI, 1.02 to 7.63). CONCLUSIONS: ECG strain identifies hypertensive patients at increased risk of developing CHF and dying as a result of CHF, even in the setting of aggressive blood pressure lowering. PMID- 16365196 TI - Disruption of the cathepsin K gene reduces atherosclerosis progression and induces plaque fibrosis but accelerates macrophage foam cell formation. AB - BACKGROUND: Cathepsin K (catK), a lysosomal cysteine protease, was identified in a gene-profiling experiment that compared human early plaques, advanced stable plaques, and advanced atherosclerotic plaques containing a thrombus, where it was highly upregulated in advanced stable plaques. METHODS AND RESULTS: To assess the function of catK in atherosclerosis, catK(-/-)/apolipoprotein (apo) E(-/-) mice were generated. At 26 weeks of age, plaque area in the catK(-/-)/apoE(-/-) mice was reduced (41.8%) owing to a decrease in the number of advanced lesions as well as a decrease in individual advanced plaque area. This suggests an important role for catK in atherosclerosis progression. Advanced plaques of catK(-/-)/apoE(-/-) mice showed an increase in collagen content. Medial elastin fibers were less prone to rupture than those of apoE(-/-) mice. Although the relative macrophage content did not differ, individual macrophage size increased. In vitro studies of bone marrow derived-macrophages confirmed this observation. Scavenger receptor mediated uptake (particularly by CD36) of modified LDL increased in the absence of catK, resulting in an increased macrophage size because of increased cellular storage of cholesterol esters, thereby enlarging the lysosomes. CONCLUSIONS: A deficiency of catK reduces plaque progression and induces plaque fibrosis but aggravates macrophage foam cell formation in atherosclerosis. PMID- 16365197 TI - Pharmacological activation of liver X receptors promotes reverse cholesterol transport in vivo. AB - BACKGROUND: Liver X receptors (LXRs) are ligand-activated transcription factors involved in the control of lipid metabolism and inflammation. Synthetic LXR agonists have been shown to inhibit the progression of atherosclerosis in mice, but the mechanism is uncertain. LXR agonism upregulates the genes encoding ATP binding cassette transporters A1 (ABCA1) and G1 (ABCG1) in macrophages, thus promoting efflux of cholesterol; it also upregulates liver and intestinal ABCG5 and ABCG8, helping to promote biliary and fecal excretion of cholesterol. Thus, LXR agonism may inhibit atherosclerosis through promotion of reverse cholesterol transport (RCT) in vivo, but this has not been proven. We previously described an in vivo method to trace the movement of cholesterol from 3H-cholesterol-labeled J774 macrophages into plasma, into liver, and ultimately into the bile and feces as free cholesterol or bile acids. In the present study we used this approach to test the hypothesis that administration of the synthetic LXR agonist GW3965 would increase the rate of macrophage RCT in vivo. METHODS AND RESULTS: Three different mouse models-wild-type C57BL/6 mice, LDLR/apobec-1 double knockout mice, and human apolipoprotein (apo)B/cholesteryl ester transfer protein (CETP) double transgenic mice-were treated with either vehicle or GW3965. Mice were injected intraperitoneally with 3H-cholesterol-labeled and cholesterol-loaded macrophages and monitored for the appearance of 3H-tracer in plasma, liver, and feces. Administration of GW3965 significantly increased the levels of 3H-tracer in plasma and feces in all 3 mouse models. CONCLUSIONS: These results demonstrate that administration of the LXR agonist GW3965 increases the rate of RCT from macrophages to feces in vivo. PMID- 16365198 TI - Standards for statistical models used for public reporting of health outcomes: an American Heart Association Scientific Statement from the Quality of Care and Outcomes Research Interdisciplinary Writing Group: cosponsored by the Council on Epidemiology and Prevention and the Stroke Council. Endorsed by the American College of Cardiology Foundation. AB - With the proliferation of efforts to report publicly the outcomes of healthcare providers and institutions, there is a growing need to define standards for the methods that are being employed. An interdisciplinary writing group identified 7 preferred attributes of statistical models used for publicly reported outcomes. These attributes include (1) clear and explicit definition of an appropriate patient sample, (2) clinical coherence of model variables, (3) sufficiently high quality and timely data, (4) designation of an appropriate reference time before which covariates are derived and after which outcomes are measured, (5) use of an appropriate outcome and a standardized period of outcome assessment, (6) application of an analytical approach that takes into account the multilevel organization of data, and (7) disclosure of the methods used to compare outcomes, including disclosure of performance of risk-adjustment methodology in derivation and validation samples. PMID- 16365199 TI - Images in cardiovascular medicine. Aortic endocarditis caused by inadvertent left ventricular pacemaker lead placement. PMID- 16365200 TI - Images in cardiovascular medicine. Infected patent ductus arteriosus. PMID- 16365201 TI - Images in cardiovascular medicine. A 360-degree aortic dissection tear mimicking massive aortic regurgitation. PMID- 16365202 TI - Letter regarding article by von Lewinski et al, "Insulin causes [Ca2+]i-dependent and [Ca2+]i-independent positive inotropic effects in failing human myocardium". PMID- 16365203 TI - Letter regarding article by Hemingway et al, "Does autonomic function link social position to coronary risk? The Whitehall II study". PMID- 16365204 TI - Retinal and cerebral microembolism during on-pump and off-pump coronary artery bypass graft surgery. PMID- 16365205 TI - Importance of pattern of alcohol consumption. PMID- 16365206 TI - Echocardiographic strain imaging for myocardial viability: an improvement over visual assessment? PMID- 16365207 TI - Retinal and cerebral microembolization during coronary artery bypass surgery: a randomized, controlled trial. AB - BACKGROUND: We sought to compare the effects on ophthalmic function of coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) and off-pump (OPCAB) grafting and to investigate whether retinal microvascular damage is associated with markers of cerebral injury. METHODS AND RESULTS: Retinal microvascular damage was assessed by fluorescein angiography and color fundus photography. Ophthalmic function was tested by the logarithm of the minimum angle of resolution visual acuity (VA), and cerebral injury, by transcranial Doppler ultrasound-detected emboli and S100 protein values. Twenty patients were randomized. Fluorescein angiography and postoperative VA could not be obtained for 1 CABG-CPB patient. Retinal microvascular damage was detected in 5 of 9 CABG CPB but in none of 10 OPCAB patients (risk difference, 55%; 95% confidence interval [CI], 23% to 88%; P=0.01). Color fundus photography detected microvascular damage in 1 CABG-CPB patient but in no OPCAB patients; this lesion was associated with a field defect, which remained after 3 months of follow-up. There was no difference in postoperative VA. Doppler high-intensity transient signals (HITS) were 20.3 times more frequent in the CABG-CPB than in the OPCAB group (95% CI, 9.1 to 45; P<0.0001). Protein S100 levels were higher in the CABG CPB than in the OPCAB group 1 hour after surgery (P<0.001). HITS were 14.7 times more frequent (95% CI, 3.5 to 62; P=0.001) and S100 level 2.1 times higher (95% CI, 1.3 to 3.5; P=0.005) when retinal microvascular damage was present. CONCLUSIONS: The relative frequency of retinal microvascular damage between groups shows the extent to which the risk of cerebral injury is reduced with OPCAB. Imaging of part of the cerebral circulation provides evidence to validate markers of cerebral injury. PMID- 16365208 TI - Binge drinking and mortality after acute myocardial infarction. AB - BACKGROUND: Moderate drinkers have a lower risk of mortality after myocardial infarction (MI). Although binge drinking has been associated with a higher risk of MI in some studies, its relation to prognosis after MI is uncertain. METHODS AND RESULTS: In a prospective, inception cohort study conducted at 45 US hospitals, 1935 patients hospitalized with a confirmed MI between 1989 and 1994 underwent detailed personal interviews. Patients reported their usual frequency of binge drinking of beer, wine, and liquor, defined as intake of 3 or more drinks within 1 to 2 hours, and were followed up for mortality for a median of 3.8 years. Of 1919 eligible patients, 250 (94% men) reported binge drinking during the prior year, and a total of 318 patients died during follow-up. Binge drinkers had a 2-fold higher risk of mortality than drinkers who did not binge (hazard ratio, 2.0; 95% confidence interval, 1.3 to 3.0). A comparison of 192 binge drinkers and 192 other patients matched on propensity scores yielded a similar result. The association between binge drinking and total mortality tended to be similar among patients whose usual alcohol intake was light or heavier and for binge drinkers who consumed beer, wine, or liquor. Usual alcohol intake was inversely associated with mortality, but binge drinking completely attenuated this relation. CONCLUSIONS: Our results suggest that alcohol consumption may be linked to potential hazards among patients who survive acute MI. Although moderate intake has been associated with lower mortality, binge drinking, even among light drinkers, appears to be associated with 2-fold higher mortality. PMID- 16365209 TI - Incremental value of strain rate analysis as an adjunct to wall-motion scoring for assessment of myocardial viability by dobutamine echocardiography: a follow up study after revascularization. AB - BACKGROUND: Assessment of myocardial viability based on wall-motion scoring (WMS) during dobutamine echocardiography (DbE) is difficult and subjective. Strain-rate imaging (SRI) is quantitative, but its incremental value over WMS for prediction of functional recovery after revascularization is unclear. METHODS AND RESULTS: DbE and SRI were performed in 55 stable patients (mean age, 64+/-10 years; mean ejection fraction, 36+/-8%) with previous myocardial infarction. Viability was predicted by WMS if function augmented during low-dose DbE. SR, end-systolic strain (ESS), postsystolic strain (PSS), and timing parameters were analyzed at rest and with low-dose DbE in abnormal segments. Regional and global functional recovery was defined by side-by-side comparison of echocardiographic images before and 9 months after revascularization. Of 369 segments with abnormal resting function, 146 showed regional recovery. Compared with segments showing functional recovery, those that failed to recover had lower low-dose DbE SR, SR increment (DeltaSR), ESS, and ESS increment (DeltaESS) (each P<0.005). After optimal cutoffs for the strain parameters were defined, the sensitivity of low dose DbE SR (78%, P=0.3), DeltaSR (80%, P=0.1), ESS (75%, P=0.6), and DeltaESS (74%, P=0.8) was better though not significantly different from WMS (73%). The specificity of WMS (77%) was similar to the SRI parameters. Combination of WMS and SRI parameters augmented the sensitivity for prediction of functional recovery above WMS alone (82% versus 73%, P=0.015; area under the curve=0.88 versus 0.73, P<0.001), although specificities were comparable (80% versus 77%, P=0.2). CONCLUSIONS: The measurement of low-dose DbE SR and DeltaSR is feasible, and their combination with WMS assessment improves the sensitivity of viability assessment with DbE. PMID- 16365210 TI - Myocardial strain analysis in acute coronary occlusion: a tool to assess myocardial viability and reperfusion. AB - BACKGROUND: This study proposes 2 new echocardiographic indices with potential application in acute coronary artery occlusion to differentiate between viable and necrotic myocardium and to identify reperfusion. We investigated whether the ratio between systolic lengthening and combined late and postsystolic shortening (L-S ratio) could identify viable myocardium and whether systolic myocardial compliance, calculated as systolic lengthening divided by systolic pressure rise, could identify necrotic myocardium. METHODS AND RESULTS: In anesthetized dogs, we measured left ventricular (LV) pressure and long-axis strain by Doppler echocardiography (SDE) and sonomicrometry. The left anterior descending coronary artery was occluded for 15 minutes with 3-hour reperfusion (n=6), for 4 hours with 3-hour reperfusion (n=6), or for 4 hours with no reperfusion (n=6). Myocardial work was quantified by pressure-segment length analysis, necrosis by triphenyltetrazolium chloride staining, and edema by water content. L-S ratio and systolic compliance were calculated by SDE. The L-S ratio ranged between 0.00 and 1.00 and was well correlated with regional myocardial work (r=0.77, P<0.0001). In entirely passive myocardium, the L-S ratio approached 1 and was similar in viable (0.88+/-0.02) and necrotic (0.81+/-0.03) myocardium. Compliance, however, was reduced in necrotic myocardium owing to edema (0.07+/-0.01%/mm Hg) but was preserved in viable myocardium (0.15+/-0.01%/mm Hg, P<0.05). Reperfusion of viable myocardium caused a reduction of the L-S ratio after 15 minutes (0.57+/ 0.06, P<0.05), reflecting recovery of function. Reperfusion of necrotic myocardium caused no change in the L-S ratio, but compliance was further reduced within 15 minutes (0.03+/-0.01%/mm Hg, P<0.05). CONCLUSIONS: Myocardial L-S ratio and compliance by SDE identified active contraction and necrosis, respectively. These indices should be tested clinically for assessment of myocardial viability and reperfusion. PMID- 16365211 TI - Comparison of valve structure, valve weight, and severity of the valve obstruction in 1849 patients having isolated aortic valve replacement for aortic valve stenosis (with or without associated aortic regurgitation) studied at 3 different medical centers in 2 different time periods. AB - BACKGROUND: Aortic valve replacement (AVR) for patients with aortic stenosis (AS) has now been available for 45 years. During this period, indications for the procedure have changed. METHODS AND RESULTS: Operatively excised stenotic aortic valves (with or without associated aortic regurgitation and without associated mitral valve disease) from 3 different medical centers (National Institutes of Health, Georgetown University Medical Center, and Baylor University Medical Center) were examined during 2 different time periods by the same physician to compare aortic valve structure, valve weight, age at operation, preoperative transvalvular peak pressure gradient, calculated aortic valve area, and whether simultaneous coronary artery bypass grafting (CABG) was performed. Compared with the first 3 decades (1961-1990) of AVR, patients having this operation during the fourth and fifth decades (1991-2004) had a lower frequency of congenitally malformed aortic valves, a higher frequency of tricuspid aortic valves, an older age, valves of lighter weight and lower transvalvular peak pressure gradients, and more often simultaneous CABG. CONCLUSIONS: Although patients having isolated AVR for AS in the present and last decade were older than in the first 3 decades of valve replacement surgery, congenitally malformed aortic valves continue to be more common than tricuspid aortic valves, but the degree of AS and therefore, valve weight was significantly lower than in the earlier decades. PMID- 16365212 TI - Diesel exhaust inhalation causes vascular dysfunction and impaired endogenous fibrinolysis. AB - BACKGROUND: Although the mechanisms are unknown, it has been suggested that transient exposure to traffic-derived air pollution may be a trigger for acute myocardial infarction. The study aim was to investigate the effects of diesel exhaust inhalation on vascular and endothelial function in humans. METHODS AND RESULTS: In a double-blind, randomized, cross-over study, 30 healthy men were exposed to diluted diesel exhaust (300 microg/m3 particulate concentration) or air for 1 hour during intermittent exercise. Bilateral forearm blood flow and inflammatory factors were measured before and during unilateral intrabrachial bradykinin (100 to 1000 pmol/min), acetylcholine (5 to 20 microg/min), sodium nitroprusside (2 to 8 microg/min), and verapamil (10 to 100 microg/min) infusions 2 and 6 hours after exposure. There were no differences in resting forearm blood flow or inflammatory markers after exposure to diesel exhaust or air. Although there was a dose-dependent increase in blood flow with each vasodilator (P<0.0001 for all), this response was attenuated with bradykinin (P<0.05), acetylcholine (P<0.05), and sodium nitroprusside (P<0.001) infusions 2 hours after exposure to diesel exhaust, which persisted at 6 hours. Bradykinin caused a dose-dependent increase in plasma tissue plasminogen activator (P<0.0001) that was suppressed 6 hours after exposure to diesel (P<0.001; area under the curve decreased by 34%). CONCLUSIONS: At levels encountered in an urban environment, inhalation of dilute diesel exhaust impairs 2 important and complementary aspects of vascular function in humans: the regulation of vascular tone and endogenous fibrinolysis. These important findings provide a potential mechanism that links air pollution to the pathogenesis of atherothrombosis and acute myocardial infarction. PMID- 16365213 TI - What a cardiologist needs to know about patients with human immunodeficiency virus infection. AB - PATIENT CASE: A 48-year-old man with human immunodeficiency virus (HIV) infection developed chronic chest pain that started after a bout of pneumonia. He has hypertension and has smoked cigarettes in the past. His current medications include Kaletra and Combivir. His total cholesterol was 331 mg/L, his HDL cholesterol was 27 mg/L, his triglycerides were 935 mg/L, and his LDL cholesterol could not be calculated. How should this patient be evaluated and managed? PMID- 16365214 TI - Acute heart failure syndromes: current state and framework for future research. PMID- 16365217 TI - Predictive models for cochlear implantation in elderly candidates. AB - OBJECTIVE: An aging American population carries a high prevalence of profound sensorineural hearing loss. We examined the performance of multichannel cochlear implant recipients in a large database of adult subjects. DESIGN: Nonconcurrent prospective study of a national cohort with multivariate regression analysis of preoperative and postoperative performance variables in multichannel cochlear implant recipients. We applied models of prediction established in previous studies to the observed results. SETTING: Referral centers with active cochlear implant programs. PATIENTS: Adolescents and adults with profound hearing loss (N = 749; age range, 14-91 years). MAIN OUTCOME MEASURE: Postoperative monosyllabic word recognition. RESULTS: The population 65 years and older demonstrated a clinically insignificant 4.6%-smaller postoperative word score compared with the population younger than 65 years. When duration of deafness exceeded 25 years, elderly recipients demonstrated higher word scores than their younger counterparts. A more significant factor affecting outcomes is the ratio of duration of deafness to age at implantation. CONCLUSIONS: Age at implantation carried relatively little predictive value for postoperative performance in subjects 65 years and older. Although a small decrement in mean speech recognition scores was evident, the clinical significance of this difference is questionable when all of the results observed in elderly patients are considered. A shorter percentage of life spent in severe-to-profound sensorineural hearing loss suggests a foundation of acoustic/auditory processing in the elderly cohort that may mitigate potential physiological effects associated with advanced age. This study confirms and extends previous observations that duration of profound deafness and residual speech recognition carry higher predictive value than the age at which an individual receives an implant. PMID- 16365218 TI - Deletion of and novel missense mutation in POU3F4 in 2 families segregating X linked nonsyndromic deafness. AB - OBJECTIVE: To analyze the physical manifestations and genetic features of 2 families segregating X-linked deafness, which is most commonly reported to be caused by mutations of the POU domain gene POU3F4 at the DFN3 locus. DESIGN: Computed tomographic study of the temporal bone in probands from each family, followed by mutation screening and deletion mapping of POU3F4 in family members. SETTING: Two midwestern genetics clinics. PARTICIPANTS: Two families with X linked deafness. MAIN OUTCOME MEASURES: Anomalies of the inner ear in the probands; results of gene mapping and severity and effects of hearing loss in the family members. RESULTS: In the first family, a large deletion was identified that includes POU3F4 and extends upstream approximately 530 kilobases; in the second family, a novel serine-to-leucine (S228L) amino acid mutation was identified in the POU-specific domain of POU3F4. Both the deletion and the missense mutation segregate with the clinical phenotype and are causally related to the deafness in these families. CONCLUSIONS: Deafness related to the POU3F4 gene is associated with dilation of the internal auditory canal and a spectrum of other temporal bone anomalies that range in severity from mild to severe dysplasia of the cochlea and semicircular canals. The consequence of these anomalies is a congenital mixed hearing loss, the sensorineural component of which progresses over time. Affected males can also present with vestibular dysfunction that is associated with delayed developmental motor milestones. Intrafamilial variability occurs. PMID- 16365219 TI - Lymphatic vessel density, nodal metastases, and prognosis in patients with head and neck cancer. AB - OBJECTIVE: To examine the relationship between intratumoral lymphatic vessel density and clinical and pathological variables in patients with head and neck squamous cell carcinoma. DESIGN: Archived paraffin-embedded biopsy specimens were sectioned and stained with hematoxylin-eosin and anti-LYVE-1 antibody, a highly specific marker for lymphatic endothelium. Tumor grade, infiltrating margin, inflammatory infiltrate, and percentage of tumor necrosis were noted and lymphatic vessel density measured using Chalkley point counting. SETTING: Tertiary care center at a university hospital. PATIENTS: A total of 168 previously untreated patients with advanced squamous cell carcinoma (73, larynx; 62, oropharynx; and 33, hypopharynx) treated with primary radiation (with or without planned neck dissection) and salvage surgery from 1992 to 1999. INTERVENTIONS: Measurement of intratumoral lymphatic vessel density in pretreatment tissue biopsy specimen. MAIN OUTCOME MEASURES: Disease-free and disease-specific survival, tumor occurrence, and nodal status. RESULTS: In patients with laryngeal carcinoma there was a significant relationship between the presence of intratumoral lymphatics and nodal metastases at presentation (P = .02) and poorly differentiated tumor grade (P = .02). Patients with high lymphatic vessel density also had a significantly worse disease-specific survival (P = .03). However, this difference was not significant with multivariate analysis. No significant relationship existed between the presence of intratumoral lymphatics and any of the clinical or pathological variables in oropharyngeal and hypopharyngeal carcinoma. CONCLUSIONS: In this patient sample, the development of intratumoral lymphatics in laryngeal carcinoma, but not in oropharyngeal or hypopharyngeal carcinoma, is associated with a spread of the tumor to regional lymph nodes. Detecting tumor lymphatic vessel proliferation is another step in the understanding of tumor biology, and the targeting of lymphatic growth may be of potential therapeutic benefit in selected patients with head and neck squamous cell carcinoma. PMID- 16365220 TI - Reliability of platelet-derived endothelial cell growth factor as a prognostic factor for oral and oropharyngeal carcinomas. AB - OBJECTIVE: To identify a strong prognostic biological marker for patients with oral and oropharyngeal squamous cell carcinomas. DESIGN: We evaluated the protein expressions of 26 tumor-associated factors, including cytokines and cytokine receptors (granulocyte colony-stimulating factor [G-CSF], interleukin 10 [IL-10], G-CSF receptor [G-CSFR], and IL-12 receptor); angiogenic factors (platelet derived endothelial cell growth factor [PD-ECGF] and vessel count); cell cycle related proteins (p27, cyclin D1, and cyclin E); apoptosis-related factors (wild type p53, Bax, Bcl-2, apoptotic index, Fas, and Fas ligand); oncogene proteins (c fos and c-Myc); cell-surface proteins (P-glycoprotein, multidrug resistance associated protein, nm23, and CD40); intracellular proteins (aryl hydrocarbon receptor nuclear translocator, aryl hydrocarbon receptor, and heat shock protein 27); and DNA mismatch-repair genes (protein encoded by human mutL homologue 1 and the human mutS homologue of the chromosome 2p gene) by means of immunohistochemical analysis. SETTING: Department of Otorhinolaryngology-Head and Neck Surgery, University of Fukui, Fukui, Japan. PATIENTS: Fifty-eight patients who underwent surgical resections of oral and oropharyngeal squamous cell carcinomas. RESULTS: A low-level PD-ECGF expression, a hypovascular count, or a low-level G-CSFR expression was associated with a favorable clinical outcome using the Kaplan-Meier method. Univariate analysis showed that PD-ECGF expression (odds, 4.19; P = .02), G-CSFR expression (odds, 4.10; P = .01), and vessel count (odds, 2.80; P = .04) had significant hazard rates. When multivariate analysis was performed on 6 factors, including sex, tumor size, lymph node metastasis, PD ECGF expression, G-CSFR expression, and vessel count, patients with a positive expression of PD-ECGF had the highest relative risk value for death due to the disease (odds, 4.94; P = .01). Also, G-CSFR was an independent prognostic indicator in the model (odds, 3.29; P = .04). No correlations between other factors and prognoses were detected. CONCLUSION: Expression of PD-ECGF was the most effective marker for making prognoses for oral and oropharyngeal squamous cell carcinomas, and G-CSFR expression was the second most effective among 26 tumor-associated factors. PMID- 16365221 TI - Combination nonviral murine interleukin 2 and interleukin 12 gene therapy and radiotherapy for head and neck squamous cell carcinoma. AB - OBJECTIVES: To demonstrate that the combination of nonviral murine interleukin (mIL) 2 and mIL-12 gene therapy and external beam radiation therapy (XRT) have an enhanced therapeutic effect for the treatment of head and neck squamous cell carcinoma (HNSCC) in an orthotopic murine model and to elucidate the mechanism of action. DESIGN: A randomized, controlled study in a murine HNSCC model. INTERVENTIONS: Tumors were established in the floor of the mouth in C3H/HeJ immunocompetent mice with the SCC VII cell line. These tumors were directly injected with single lipid-formulated mIL-2 or single polymer-formulated mIL-12 or a combination of them and with phosphate-buffered saline or vector without mIL 2 and mIL-12 gene as controls. Then the local tumor was radiated twice with a dose of 1 Gy the next day and injected again 4 days later. Antitumor responses, cytokine expression, and natural killer cell and cytolytic T-lymphocyte activity were assayed. Meanwhile, tumor sizes were measured before and after treatment and compared among the different treatment groups and the controls. RESULTS: The combination mIL-2 + mIL-12 + XRT demonstrated a significant increase in antitumor effects compared with single therapy or controls. Increased expression levels of primary and secondary cytokines were found in the group treated with mIL-2 + mIL 12, and this effect was preserved when mIL-2 and mIL-12 treatments were combined with XRT. Combination therapy significantly increased antitumor effects, T lymphocyte infiltration of CD4(+)and CD8(+), and the numerous necroses compared with monotherapy. CONCLUSIONS: Combination mIL-2 and mIL-12 gene therapy and XRT generates potent antitumor immune responses against HNSCC and significantly increases necrosis (apoptosis) in an orthotopic murine model of HNSCC. The nonviral mIL-2 and mIL-12 gene delivery system was well tolerated. Further optimization of treatment strategy for patients with HNSCC is warranted as well as consideration for human clinical trials. PMID- 16365222 TI - Idiopathic vocal cord palsies and associated neurological conditions. AB - OBJECTIVE: To retrospectively review the clinical case records of patients with idiopathic vocal cord palsies (VCPs) for the presence of preexisting or subsequent development of neurological disease, including multiple sclerosis, motor neuron disease, myasthenia gravis, cerebrovascular disease, and Guillain Barre syndrome. DESIGN: Retrospective case review of all patients with VCP presenting sequentially within a 45-month time span. SETTING: Tertiary referral center. PATIENTS: One hundred ninety-three patients with VCP. RESULTS: Thirty five cases of VCP (18.1%) were idiopathic. Eight (22.8%) resolved after a mean time of 5 months. A preexisting central nervous system condition was noted in 9 (25.7%) of 35 patients with idiopathic VCP. A subsequent central nervous system condition developed in 7 patients (20.0%). These included 2 cases of cerebrovascular accidents, 1 case of postpolio syndrome with respiratory failure, and 1 case of polyneuropathy secondary to paraneoplastic syndrome. CONCLUSIONS: A high frequency of neurological conditions was observed in adult patients initially presenting with idiopathic VCP. Patients with VCP but without overt neurological disease may also subsequently develop a serious neurological condition. Careful neurological evaluation of all patients with idiopathic VCP is recommended. PMID- 16365223 TI - Fibroblast transplantation in the airway: implications for subglottic stenosis. AB - OBJECTIVE: Because subglottic stenosis (SGS) represents one of the most challenging pathologies confronting the pediatric otolaryngologist, our laboratory is investigating the role fibroblasts play in mucosal scar formation in the course of SGS development. Our objective is to establish cell transplantation into the subglottic mucosal wound bed as a viable tool for examining the cellular processes that underlie the development of SGS. DESIGN: A series of 2 animal experiments, with animals assigned to a control, vehicle-only, or cell-treated group. SETTING: John G. Rangos Sr Research Center, Children's Hospital of Pittsburgh, Pittsburgh, Pa. SUBJECTS: Twenty-six New Zealand white rabbits. This animal model has been well established in the study of SGS formation. INTERVENTIONS: Fluorescently labeled exogenous fibroblasts were transplanted into the injured subglottis of the rabbits. RESULTS: Exogenous fibroblasts derived from fetal and adult dermis and subglottic mucosa were successfully transplanted into the injured subglottic mucosa of adult rabbits. Transplanted fibroblasts survived into the latter stages of wound healing (at 14 and 21 days) and appeared to be associated with a mild inflammatory cell influx and active remodeling of the mucosal wound bed. CONCLUSION: Cell transplantation is a viable tool for the study of fibroblast activity in the mucosal wound bed. PMID- 16365224 TI - Dornase alfa as postoperative therapy in cystic fibrosis sinonasal disease. AB - OBJECTIVE: To determine the benefit of nasally inhaled dornase alfa in patients with cystic fibrosis and nasal symptoms. DESIGN: Double-blind placebo-controlled trial. SETTING: Cystic Fibrosis Regional Center of Campania at the University of Naples "Federico II." PATIENTS: A total of 24 patients with cystic fibrosis and chronic sinusitis. INTERVENTIONS: Patients underwent sinonasal surgery during a 3 year period and received once-daily doses of either dornase alfa (2.5 mg) or hypotonic saline solution (5 mL) beginning 1 month after surgery and for a 12 month period. MAIN OUTCOME MEASURES: Primary outcomes were nasal-related symptoms and nasal endoscopic appearance; secondary outcomes were forced expiratory volume in 1 second, nasal computed tomography findings, and saccharine clearance test results. Patients were evaluated before and after treatment. RESULTS: After surgery, all outcomes were significantly improved for each treatment at 1 month (P<.05); primary outcomes were improved at 24 and 48 weeks in the group receiving dornase alfa (P<.05), and at 12 weeks in the group receiving placebo. Secondary outcomes were better in the dornase alfa group (P<.01) than in the placebo group at 12 months except for the saccharine clearance test results. In particular, median relative difference in forced expiratory volume in 1 second between dornase alfa and placebo was significantly improved in the dornase alfa group (P<.01). CONCLUSIONS: Nasally inhaled dornase alfa can be effective in patients with cystic fibrosis and sinonasal disease who do not respond to conventional therapy after surgical treatment. Further studies should be carried out to determine the long-term effect on sinus disease, recurrence of polyps, and quality of life. PMID- 16365225 TI - Aerodynamic effects of inferior turbinate reduction: computational fluid dynamics simulation. AB - OBJECTIVE: To investigate the aerodynamic consequences of conservative unilateral inferior turbinate reduction using computational fluid dynamics methods to accomplish detailed nasal airflow simulations. DESIGN: A high-resolution, finite element mesh of the nasal airway was constructed from magnetic resonance imaging data of a healthy man. Steady-state, inspiratory airflow simulations were conducted at 15 L/min using the techniques of computational fluid dynamics. INTERVENTION: Circumferential removal of 2 mm of soft tissue bulk along the length of the left inferior turbinate was modeled. MAIN OUTCOME MEASURES: Nasal airflow distribution and pressure profiles were computed before and after simulated left inferior turbinate reduction. RESULTS: Simulated inferior turbinate reduction resulted in a broad reduction of pressure along the nasal airway, including the regions distant from the inferior turbinate vicinity. In contrast, relative airflow changes were regional: airflow was minimally affected in the valve region, increased in the lower portion of the middle and posterior nose, and decreased dorsally. CONCLUSION: Use of computational fluid dynamics methods should help elucidate the aerodynamic significance of specific surgical interventions and refine surgical approaches to the nasal airway. PMID- 16365226 TI - Up-regulation of surfactant protein A in chronic sialadenitis. AB - OBJECTIVES: Salivary secretions play a critical role in maintaining the health of the oral cavity, which is the first gate of entry to the airways and thus is exposed to a variety of environmental insults. Surfactant protein A (SP-A) is a member of the collectin family and plays an important role in first-line airway defense. The objectives of this study were to examine the expression of SP-A messenger RNA and protein in human salivary glands and to investigate its up regulation during inflammatory conditions. DESIGN: Reverse transcription polymerase chain reaction was performed on salivary gland tissues from patients and a control group. The expression levels of SP-A to GAPDH (glyceraldehyde-3 phosphate dehydrogenase) transcripts were semiquantified by densitometry. We also characterized the cellular localizations of SP-A protein immunohistochemically. SETTING: Tertiary academic institution. PATIENTS: Ten patients with chronic sialadenitis and 10 patients with healthy salivary glands. RESULTS: Surfactant protein A messenger RNA and protein were detected in glands of patients who were healthy and in those with chronic sialadenitis. The expression levels of SP-A messenger RNA in the salivary glands of patients with chronic sialadenitis was significantly increased compared with those in healthy salivary glands. Immunohistochemical staining revealed SP-A immunoreactivity in the ductal epithelia of healthy salivary glands and in the salivary glands of those with chronic sialadenitis, and stronger immunoreactivity was observed in those with chronic sialadenitis tissues. CONCLUSIONS: Surfactant protein A is present in the salivary gland epithelium and is up-regulated in individuals with chronic sialadenitis. These results suggest that salivary gland SP-A may play an important role in the innate host defense of human salivary glands. PMID- 16365228 TI - Radiology quiz case 1. Benign osteoblastoma. PMID- 16365227 TI - Foregut duplication cyst of the hypopharynx. PMID- 16365229 TI - Radiology quiz case 2. Cutaneous dental sinus tract. PMID- 16365230 TI - Pathology quiz case 1. Adenocarcinoma with features of mucinous adenocarcinoma (MAC) and cystadenocarcinoma (CAC). PMID- 16365231 TI - Pathology quiz case 2. Nodular fasciitis. PMID- 16365233 TI - The Department of Surgery, University of California, San Francisco. PMID- 16365234 TI - Resection and primary anastomosis is a valid surgical option for infants with necrotizing enterocolitis who weigh less than 1000 g. AB - HYPOTHESIS: Primary anastomosis following intestinal resection is a valid surgical option in the treatment of infants with necrotizing enterocolitis (NEC) who weigh less than 1000 g. DESIGN: Retrospective case series. SETTING: Tertiary neonatal surgery referral center. PATIENTS: All infants with confirmed NEC weighing less than 1000 g admitted to our intensive care unit over 4 years. INTERVENTION: For infants requiring laparotomy and intestinal resection, primary anastomosis was performed whenever the clinical condition permitted. MAIN OUTCOME MEASURES: Short- and long-term survival, length of intensive care unit stay, and complications. RESULTS: Fifty-one infants with NEC who weighed less than 1000 g were admitted during the study period. Twelve infants underwent intestinal resection and primary anastomosis (median weight at surgery, 0.83 kg; range, 0.6 0.96 kg). One infant developed recurrent NEC, requiring further surgery, but there were no anastomotic leakages and no strictures. The median postoperative stay on our intensive care unit was 14 days (range, 2-49 days). All 12 infants survived their episode of acute NEC, and 8 are alive, with a median follow-up of 34.2 months (range, 4.7-48.4 months). Only 1 death was related to NEC. During the same period, 14 infants underwent intestinal resection and stoma formation. Ten survived the acute episode, and 6 are alive at a median follow-up of 24.0 months (range, 13.1-33.9 months). The median postoperative intensive care unit stay was 18 days (range, 2-74 days). Necrotizing enterocolitis-related complications occurred in 8 of these infants. CONCLUSIONS: The outcome of infants with NEC who weigh less than 1000 g and undergo primary anastomosis is comparable to that of infants treated using alternative techniques. It is, therefore, a valid surgical option in selected infants. The mortality in this group of infants is high. PMID- 16365235 TI - Reconstructive breast implantation after mastectomy for breast cancer: clinical outcomes in a nationwide prospective cohort study. AB - BACKGROUND: Clinical reports have raised concern about local complications following breast implantation used in reconstructive or cosmetic surgery, but there is a shortage of epidemiological studies in this area. OBJECTIVE: To assess in a prospective epidemiological manner the occurrence of short-term local complications in a nationwide implantation registry. DESIGN, SETTING, AND PARTICIPANTS: The Danish Registry for Plastic Surgery of the Breast prospectively collects preoperative, perioperative, and postoperative information on Danish women undergoing breast augmentation. Through the registry, we collected data on short-term local complications among 574 women who underwent postmastectomy reconstruction with breast implants from June 1, 1999, through July 24, 2003. MAIN OUTCOME MEASURES: Complication incidence rates. RESULTS: Thirty-one percent of the women who underwent initial implantation developed at least 1 adverse event. Forty-nine percent of the adverse events occurred within 3 months after implantation and 67% within 6 months. Surgical intervention was required after initial implantation among 21% of women, most frequently because of capsular contracture, asymmetry, or displacement of the implant. Thirty-six percent of women who underwent subsequent implantation experienced at least 1 adverse event, and 21% underwent surgical intervention to treat definitive complications or to optimize cosmetic result. CONCLUSIONS: Women who undergo postmastectomy breast implantation frequently experience short-term local complications. Surgical or medical intervention is commonly required during the reconstructive course, but reconstruction failure (loss of implant) is rare. However, when weighing benefits and risks associated with reconstruction, the patient should consider that breast reconstruction is a process involving planned and unplanned supplementary surgical correction to achieve the desired result. PMID- 16365236 TI - Small-bowel diaphragm disease: seven surgical cases. AB - HYPOTHESIS: Small-bowel diaphragm disease is an important source of gastrointestinal tract bleeding and subacute intestinal obstruction that may require surgical intervention. DESIGN: Case series. SETTING: Tertiary-care academic medical center. PATIENTS: Seven consecutive patients with histologically confirmed enteropathy induced by use of nonsteroidal anti-inflammatory drugs received treatment at our institution from February 2001 to February 2004. INTERVENTIONS: Laparotomy with small-bowel resection. MAIN OUTCOME MEASURES: Initial symptoms and signs, findings at diagnostic workup, type of medication and duration of use, operation performed, and intraoperative findings were evaluated. RESULTS: Patients were identified who had symptoms of subacute intestinal obstruction or signs of gastrointestinal tract bleeding and were determined to have used nonsteroidal anti-inflammatory drugs for differing periods. All 7 patients underwent extensive nondiagnostic radiologic and endoscopic examinations. The diagnosis of diaphragm disease was ultimately made at either video capsule endoscopy or laparotomy. Randomly distributed diaphragms throughout the jejunum and ileum necessitated resection in all cases. No patient has had a documented recurrence, although follow-up is short (range, 0-20 months). CONCLUSIONS: Nonsteroidal anti-inflammatory drugs are clearly linked to pathologic findings of diaphragm disease in both the upper and lower gastrointestinal tracts. Although rarely reported in the surgical literature, small-bowel diaphragm disease may be more common than thought and can manifest as gastrointestinal tract bleeding or obstruction. Diagnosis is difficult and may require laparotomy and small-bowel resection. PMID- 16365237 TI - Impact of surgical treatment on respiratory muscle dysfunction in symptomatic hyperparathyroidism. AB - HYPOTHESIS: We hypothesized that surgical treatment would improve respiratory muscle strength in symptomatic hyperparathyroidism (HPT). DESIGN: Prospective clinical trial. SETTING: A tertiary referral center. PATIENTS: Fifteen consecutive patients with symptomatic HPT and 10 with euthyroid multinodular goiter (control group) without a history of obstructive or restrictive lung disease. INTERVENTIONS: Forced vital capacity and forced expiratory volume in 1 second were measured before and 6 months after surgery to estimate respiratory muscle involvement. These measurements were compared with the reference values estimated individually in each patient. Mann-Whitney and Wilcoxon signed rank tests were used for statistical analysis, and P<.05 was considered statistically significant. MAIN OUTCOME MEASURES: Respiratory dysfunction in patients with symptomatic HPT, pulmonary function after parathyroidectomy, and the correlation between the preoperative serum parathyroid hormone and total serum calcium values and the impairment in pulmonary function. RESULTS: Preoperative forced vital capacity and forced expiratory volume in 1 second measurements were below the reference values in 11 (73%) and 9 (60%) patients, respectively. All the patients were normocalcemic, and forced vital capacity and forced expiratory volume in 1 second measurements significantly improved at postoperative month 6 (P = .001). No significant difference was detected in the control group. Improvement in pulmonary function correlated with preoperative serum calcium and parathyroid hormone values in patients with HPT (P<.05 and P<.001, respectively). CONCLUSIONS: Symptomatic HPT impairs inspiratory and expiratory components of respiratory function, and normalization of serum calcium levels after surgical treatment is associated with a significant improvement in lung function. PMID- 16365238 TI - Patterns of recurrence in patients with melanoma after radical lymph node dissection. AB - HYPOTHESIS: Previous interventions (excisional biopsy, incomplete dissection) in the regional basin that drain a melanoma site prior to definitive surgical procedures significantly increase the risk of melanoma recurrence in the surgical field. DESIGN: Retrospective analysis. SETTING: Tertiary care referral center. PATIENTS: One hundred forty-one consecutive patients who underwent radical lymph node dissection (RLND) either in the groin or the axilla owing to malignant melanoma were followed up for a median period of 41 months. INTERVENTIONS: All of the 141 patients received either elective or therapeutic RLND. Their medical records were analyzed for demographic data, disease history, previous treatments, recurrence patterns, and survival. MAIN OUTCOME MEASURES: Patterns of first recurrence after RLND and survival. RESULTS: Radical lymph node dissection was performed on 148 lymph node basins (141 patients; 86 axillae and 62 groins). Nineteen patients (13%) received previous open interventions in the lymph node basin (tampering) other than radical dissection. Radical lymph node dissection was performed prophylactically in 38 basins (26%), for palpable disease in 75 (51%), and for a positive sentinel node in 35 (24%). There were 74 failures (52%) of RLND: 51 patients (70%) with systemic disease, 12 (16%) with recurrence in the surgical field, 9 (11%) with in-transit metastases, and 2 (3%) with local recurrence. On multivariate analysis, the only significant predictors of recurrence after RLND were Breslow thickness of greater than 4 mm (P = .02), tampering (P = .01), and lymph node capsular invasion (P = .001). Tampering was the only independent prognosticator of failure in the surgical field, as tampering was noted in 10 (83%) of 12 patients with failure in the surgical field as compared with 6 (10%) of 62 patients with other types of first failures (P<.001). This effect did not translate into a survival difference (P = .54). Failure in the surgical field was not detected in any of the patients who underwent sentinel lymph node biopsy. CONCLUSIONS: Previous interventions (excisional biopsy, incomplete dissection) in the regional basin that drain a melanoma site prior to definitive surgical procedures significantly increase the risk of melanoma recurrence in the surgical field, and they should be avoided. Fine-needle aspiration and sentinel node biopsy, performed with strict surgical oncologic techniques, are safe with regard to failure in the surgical field. PMID- 16365239 TI - Advantages of mini-laparoscopic vs conventional laparoscopic cholecystectomy: results of a prospective randomized trial. AB - HYPOTHESIS: The use of smaller instruments during laparoscopic cholecystectomy (LC) has been proposed to reduce postoperative pain and improve cosmesis. However, despite several recent trials, the effects of the use of miniaturized instruments for LC are not well established. We hypothesized that LC using miniports (M-LC) is safe and produces less incisional pain and better cosmetic results than LC performed conventionally (C-LC). DESIGN: A patient- and observer blinded, randomized, prospective clinical trial. SETTING: A tertiary care, university-based hospital. PATIENTS: Seventy-nine patients scheduled for an elective LC who agreed to participate in this trial were randomized to undergo surgery using 1 of the 2 instrument sets. The criteria for exclusion were American Society of Anesthesiologists class III or IV, age older than 70 years, liver or coagulation disorders, previous major abdominal surgical procedures, and acute cholecystitis or acute choledocholithiasis. INTERVENTION: Laparoscopic cholecystectomy performed with either conventional or miniaturized instruments. MAIN OUTCOME MEASURES: Patients' age, sex, operative time, operative blood loss, intraoperative complications, early and late postoperative incisional pain, and cosmetic results. RESULTS: Thirty-three C-LCs and 34 M-LCs were performed and analyzed. There were 8 conversions (24%) to the standard technique in the M-LC group. No intraoperative or major postoperative complications occurred in either group. The average incisional pain score on the first postoperative day was significantly less in the M-LC group (3.9 vs 4.9; P = .04). No significant differences occurred in the mean scores for pain on postoperative days 3, 7, and 28. However, 90% of patients in the M-LC group and only 74% of patients in the C LC group had no pain (visual analog scale score of 0) at 28 days postoperatively (P = .05). Cosmetic results were superior in the M-LC group according to both the study nurse's and the patients' assessments (38.9 vs 28.9; P<.001, and 38.8 vs 33.4; P = .001, respectively). CONCLUSIONS: Laparoscopic cholecystectomy can be safely performed using 10-mm umbilical, 5-mm epigastric, 2-mm subcostal, and 2-mm lateral ports. The use of mini-laparoscopic techniques resulted in decreased early postoperative incisional pain, avoided late incisional discomfort, and produced superior cosmetic results. Although improved instrument durability and better optics are needed for widespread use of miniport techniques, this approach can be routinely offered to many properly selected patients undergoing elective LC. PMID- 16365240 TI - Significance of cellular distribution of ezrin in pancreatic cystic neoplasms and ductal adenocarcinoma. AB - HYPOTHESIS: Pancreatic cystic neoplasms and ductal adenocarcinoma manifest diverse clinical features and prognoses, which might be related to cellular distribution of ezrin modulated through various trophic molecules. DESIGN: Laboratory investigation and retrospective analysis. SETTING: Medical school affiliated university hospital. PATIENTS AND METHODS: Patients with solid pseudopapillary tumor (SPT) (n = 12), mucinous cystic neoplasm (MCN) (n = 18), intraductal papillary mucinous tumor (IPMT) (n = 18), and ductal adenocarcinoma (PA) (n = 73) of the pancreas were studied. Expression of epidermal growth factor (EGF) and its receptor (EGFR) and ezrin were determined using immunohistochemistry. Epidermal growth factor receptor and ezrin expression in sodium butyrate (SB)-treated PA cell line PANC-1 was determined using immunocytochemistry. Messenger RNA expression of ezrin in the PANC-1 cell line treated with SB was determined using reverse transcriptase-polymerase chain reaction. Multivariate analysis of survival of patients with PA was performed. RESULTS: None of 12 SPTs displayed synchronous expression of EGF and EGFR, while all 3 malignant SPTs displayed membranous ezrin expression. One of 18 MCNs displayed synchronous expression of EGF and EGFR, while 4 of 6 borderline malignant and 8 of 8 malignant MCNs displayed membranous ezrin expression. Two of 4 borderline malignant and 11 of 11 malignant IPMTs displayed synchronous expression of EGF and EGFR, and all borderline malignant and malignant IPMTs displayed membranous ezrin expression. Less differentiated PA displayed EGF, EGFR, and membranous ezrin expression more frequently compared with more differentiated PA. Epidermal growth factor receptor expression of PANC-1 cells decreased in an SB dose dependent manner, in which PANC-1 cells became more differentiated and membranous ezrin expression of PANC-1 cells decreased correspondingly. Messenger RNA expression of ezrin in PANC-1 cells also decreased in an SB dose dependent manner. Patients with PA with membranous ezrin expression had a poorer prognosis compared with those without (P = .02). CONCLUSIONS: Membranous translocation of ezrin might play a role during malignant transformation of SPT, MCN, IPMT, and PA, which are either dependent on (IPMT and PA) or independent of (SPT and MCN) the EGF-EGFR pathway. Membranous ezrin expression represents a prognostic factor for PA. PMID- 16365241 TI - Effect of surgeon and hospital characteristics on outcome after pyloromyotomy. AB - BACKGROUND: Previous studies have suggested that the outcome after pyloromyotomy is improved with increased surgeon experience. Others have proposed that infants with pyloric stenosis are best treated by specialty-trained pediatric surgeons or at children's hospitals. HYPOTHESIS: Surgeon and hospital characteristics affect complications, length of stay, and hospital charges after pyloromyotomy. DESIGN: Data for a nationally representative sample of infants (n = 1277) who underwent pyloromyotomy in 2000 in the United States were obtained from the Kids' Inpatient Database. Surgeon and hospital volumes were stratified into quintiles. Multivariate analyses were performed to analyze the impact of surgeon and hospital volume on length of stay, charges, and major operative complications using models that accounted for the hierarchical structure of patient-, surgeon-, and hospital-level covariates. RESULTS: No association between surgeon volume and either length of stay or charges was observed. Higher surgeon volume, however, was associated with fewer complications (P<.001). Surgeons with the highest volume had a 90% lower risk of complications than those with the lowest volume. Higher hospital volume was associated with shorter length of stay (P<.001). No association between hospital volume and either charges or risk of complications was observed. CONCLUSIONS: Higher surgeon and hospital volumes are associated with better outcome among infants who are treated for pyloric stenosis. Identification of aspects of medical and surgical treatment that account for this finding may lead to improvement in the outcome of infants undergoing pyloromyotomy. PMID- 16365242 TI - Accelerated growth of bariatric surgery with the introduction of minimally invasive surgery. AB - HYPOTHESIS: An increase in national utilization of bariatric surgery correlates with the dissemination of laparoscopic bariatric surgery. DESIGN: Evaluation of Nationwide Inpatient Sample data from 1998 through 2002. SETTING: National database. PATIENTS: A total of 188,599 patients underwent bariatric surgery for the treatment of morbid obesity. MAIN OUTCOME MEASURES: Annual total number of bariatric operations, the proportion of Roux-en-Y gastric bypass vs gastroplasty, the proportion of laparoscopic cases, postoperative length of stay, crude in hospital mortality, and the number of institutions that perform bariatric surgery. RESULTS: Between 1998 and 2002, the number of bariatric operations increased from 12,775 cases to 70,256 cases. The rate of bariatric surgery increased from 6.3 to 32.7 procedures per 100,000 adults. Laparoscopic bariatric surgery increased from 2.1% to 17.9%. The number of bariatric surgeons with membership in the American Society for Bariatric Surgery increased from 258 to 631, and the number of institutions that perform bariatric surgery increased from 131 to 323. During this 5-year period, the annual rate of laparoscopic bariatric surgery increased exponentially (by 44-fold) compared with a linear growth in open bariatric surgery (by 3-fold). CONCLUSIONS: Between 1998 and 2002, there was a 450% increase in the number of bariatric operations performed in the United States, a 144% increase in the number of American Society for Bariatric Surgery bariatric surgeons, and a 146% increase in the number of bariatric centers. The growth of laparoscopic bariatric surgery during this 5-year period greatly exceeds that of open bariatric surgery. PMID- 16365243 TI - Association of manganese superoxide dismutase expression with progression of carcinogenesis in Barrett esophagus. AB - HYPOTHESIS: The down-regulation of manganese superoxide dismutase (MnSOD) expression plays a role in the progression of Barrett esophagus (BE). DESIGN: An evaluation of 92 esophageal samples, including 17 patients with normal esophagus, 22 with intestinal metaplasia, 22 with indefinite/low-grade dysplasia, 16 with high-grade dysplasia (HGD), and 15 with esophageal adenocarcinoma were evaluated for MnSOD expression. We evaluated MnSOD expression using immunohistochemistry and graded it separately on a 2-category ordinal scale in relation to the mucosa and submucosa that ranged from 0 (no staining) to 3 (strong staining). The total grading score of MnSOD immunoreactivity was the addition of mucosa and submucosa intensity, from 0 (no immunoreactivity in any of the anatomic sites) to a maximum score of 6 (strong staining reaction in both of the histoanatomic sites). SETTING: Study subjects were recruited from the Barrett's Esophageal Registry at the University of Louisville, Louisville, KY. MAIN OUTCOME MEASURE: Manganese superoxide dismutase expression in established groups of progressive BE. RESULTS: Ninety-two samples were evaluated for MnSOD expression. The expression of MnSOD was found to be significantly reduced in samples with specialized intestinal metaplasia (mean score, 1.8), low-grade dysplasia (mean, 2.2), high-grade dysplasia (mean, 2.4), and esophageal adenocarcinoma (mean, 2.4) when compared with normal esophagus (mean, 3.9; P = .002). Manganese superoxide dismutase expression was similar for esophageal adenocarcinoma and high-grade dysplasia. CONCLUSIONS: Manganese superoxide dismutase expression is significantly reduced in patients with BE with high-grade dysplasia and esophageal adenocarcinoma. Manganese superoxide dismutase is related to the progression of BE and may represent one of the primary factors in oxidative stress protection. Further evaluation within genotypic expression and the role of antioxidants is needed in the effective screening and treatment of BE. PMID- 16365244 TI - Preexisting morbidity as an independent risk factor for perioperative acute thromboembolism syndrome. AB - HYPOTHESIS: Preexisting morbidities are risk factors for perioperative arterial or venous thromboembolic events and subsequent death within 30 postoperative days. DESIGN: Prospective cohort study. SETTING: University-affiliated general hospital. PATIENTS: A total of 21,903 surgery patients treated from January 1, 1991, through December 31, 2002. MAIN OUTCOME MEASURES: Independent risk factors for perioperative arterial or venous thromboembolic events. RESULTS: History of atrial fibrillation and coronary artery disease increased the risk of myocardial infarction (odds ratio [95% confidence interval], 4.3 [2.8-6.7]). History of stroke increased the risk of stroke (2.4 [1.4-4.1]) and death (4.7 [1.3-17.3]). Diabetes mellitus increased the risk of myocardial infarction (2.1 [1.3-3.2]), and hyperuricemia increased the risk of stroke (3.5 [1.2-9.8]), and both increased the risk of death (4.3 [1.3-14.1] and 11.8 [2.2-63.5], respectively). History of myocardial infarction increased the risk of deep vein thrombosis (7.7 [1.7-34.7]). Cancer increased the risk of all thromboembolism (2.4 [1.9-3.2]). Trend analysis showed that preexisting morbidities will increase 1.5-fold and thromboembolic events will increase 3-fold during the next decade. CONCLUSION: Cardiac and cerebrovascular diseases, metabolic diseases, and cancer are becoming increasingly high-risk comorbidities for perioperative acute thromboembolism syndrome. PMID- 16365245 TI - Transanal endoscopic repair of rectal anastomotic defect. AB - Surgeons often encounter difficulty when constructing a colorectal anastomosis in the "hostile pelvis." Examples include performing low anterior resection or colostomy takedown in the setting of prior radiation, severe inflammation, or a narrow pelvis. Circular staplers have made low anastomosis a viable alternative to permanent colostomy in these situations. However, the surgeon may occasionally be faced with the difficult decision of how to manage a gross disruption of a stapled anastomosis in a pelvis that will not permit anastomotic redo. The traditional approach to this would be creating a permanent colostomy. We describe an alternate approach: endoscopic suturing with protecting ileostomy. We have successfully applied this technique to 4 patients with gross anastomotic disruption in a hostile pelvis. All patients tolerated the procedure well and have maintained normal bowel function without the need for a permanent colostomy. PMID- 16365246 TI - Image of the month. Solid pseudopapillary tumor of the pancreas. PMID- 16365247 TI - Anastomosis of Riolan revisited: the meandering mesenteric artery. AB - The eponym anastomosis of Riolan suggests that Jean Riolan (1580-1657), a famous 17th century French anatomist, was the first to describe this mesenteric arterial connection between the superior and inferior mesenteric arteries. Riolan was a strong defender of traditional Galenic doctrine in medicine and proved a vigorous opponent of the new concept of the circulation of blood as exposed by William Harvey (1578-1657). As confirmed by examining his anatomy book published in 1649, it is unlikely that Riolan would have conceived an arterial collateral pathway in the mesocolon. He probably had observed vascular arcades running along the inner border of the colon. It was not until 1743 that Albrecht von Haller (1708-1777) gave a detailed description of the anatomy of the mesenteric arteries, referring to the arterial collateral connection between the superior and inferior mesenteric arteries as the Arcus Riolani in honor of an old master of anatomy. PMID- 16365248 TI - Primary retroperitoneal abscess extending to the calf. AB - A 77-year-old man with diabetes mellitus presented with a 2-month history of lumbago radiating to the right lower limb as well as high fever spikes. Physical examination revealed a distended abdomen with right lower quadrant tenderness. A computed tomographic scan of the abdomen revealed a large right retroperitoneal cavity containing an air-fluid level that was consistent with a gas-producing abscess. The patient began receiving intravenous antibiotics, but fever and abdominal pain persisted and a large, fluctuating, tender swelling appeared on the medial aspect of his right thigh and right calf. The patient underwent surgical exploration: a right lateral abdominal incision was performed, and the pus collection in retroperitoneal space was completely evacuated. We also made 3 separate incisions on the medial aspect of the right thigh and 1 incision on the upper calf, resulting in the drainage of pus. The patient made a slow but steady recovery. PMID- 16365250 TI - Efficacy of gabapentin in the management of pruritus of unknown origin. PMID- 16365251 TI - Alefacept for alopecia areata. PMID- 16365252 TI - Elimination of a community-acquired methicillin-resistant Staphylococcus aureus infection in a nurse with atopic dermatitis. PMID- 16365253 TI - Impact of obesity and smoking on psoriasis presentation and management. AB - OBJECTIVE: To study the impact of obesity and smoking on psoriasis. DESIGN: Cross sectional study. SETTING: University of Utah Department of Dermatology clinics. PATIENTS: A case series of patients with psoriasis enrolled in the prospective Utah Psoriasis Initiative (UPI) (which carefully performs phenotyping of patients with psoriasis) was compared with 3 population databases: the Behavioral Risk Factor Surveillance System of the Utah population, the 1998 patient-member survey from the National Psoriasis Foundation, and 500 adult patients who attend our clinics and do not have psoriasis (non-psoriatic population). RESULTS: The prevalence of obesity in patients within the UPI population was higher than that in the general Utah population (34% vs 18%; P<.001) and higher than that in the non-psoriatic population attending our clinics. Assessment of body image perception with a standardized diagram in the UPI group resulted in the median body image score of normal weight at 18 years of age and the onset of psoriasis, but it changed to overweight at the time of enrollment in the UPI. Thus, obesity appears to be the consequence of psoriasis and not a risk factor for onset of disease. We did not observe an increased risk for psoriatic arthritis in patients with obesity; furthermore, obesity did not positively or negatively affect the response or the adverse effects of topical corticosteroids, light-based treatments, and systemic medications. The prevalence of smoking in the UPI population was higher than in the general Utah population (37% vs 13%; P<.001) and higher than in the non-psoriatic population (37% vs 25%; P<.001). We found a higher prevalence of smokers in the obese population within the UPI than in the obese population within the Utah population (25% vs 9%; P<.001). CONCLUSIONS: Patients with psoriasis attending the University of Utah Dermatology Clinics were more likely to be obese and to smoke compared with non-psoriatic patients and more likely to be obese compared with other large cohorts with psoriasis. Smoking appears to have a role in the onset of psoriasis, but obesity does not. The high prevalence of obesity and smoking in a psoriasis cohort has not been previously noted; if confirmed, it supports the prediction that a significant portion of patients with psoriasis will have the comorbid conditions and public health issues of those with obesity and smoke. PMID- 16365254 TI - Prevalence and treatment of psoriasis in the United Kingdom: a population-based study. AB - OBJECTIVE: To measure the prevalence and treatment of psoriasis in the United Kingdom. DESIGN: Cross-sectional study to determine prevalence and cohort study to determine treatment patterns. SETTING: Outpatient practices of general practitioners. PATIENTS: We included in the analysis all patients who were registered with a general practitioner in the General Practice Research Database from 1987 to 2002. MAIN OUTCOME MEASURES: The prevalence and treatment of psoriasis. RESULTS: We identified 114 521 patients with psoriasis of a total population of 7 533 475 patients, yielding a prevalence of 1.5%. The prevalence of psoriasis increases more rapidly in young female patients compared with young male patients and declines significantly in patients 70 years and older, regardless of sex. Overall, 91.8% of patients with a diagnosis of psoriasis received a prescription for psoriasis treatment on or after the date of their first diagnostic code of psoriasis in the General Practice Research Database. Most of the patients (55.2%) received only 1 or 2 prescriptions for psoriasis in the first year after psoriasis was documented in the General Practice Research Database. CONCLUSIONS: The epidemiology of psoriasis in the General Practice Research Database population is similar to that of other epidemiologic studies of psoriasis performed in the United Kingdom, the United States, and other Western countries. Psoriasis carries a substantial burden given its high prevalence and its associated need for prescription therapy. Additional studies are necessary to determine why the prevalence of psoriasis increases more rapidly in female patients and to determine why the prevalence decreases in patients 70 years and older. PMID- 16365255 TI - Targeted UV-B phototherapy for plaque-type psoriasis. AB - OBJECTIVE: To determine whether targeted UV-B phototherapy is efficacious and safe in the treatment of localized psoriasis and whether there is a dose-response relationship. DESIGN: Randomized, evaluator-blind, controlled study. SETTING: Dermatology clinic in a large university-based hospital in Bangkok, Thailand. PATIENTS: Fourteen patients with stable, localized, plaque-type psoriasis. INTERVENTIONS: Patients were randomized to receive different fluences of targeted UV-B phototherapy 3 times weekly based on predetermined minimal erythema doses (MEDs). Treatment fluences were constant throughout the study period of 4 weeks. Follow-up was carried out until lesions returned to original state. MAIN OUTCOME MEASURES: Modified psoriasis area and severity index. RESULTS: All fluences of UV B produced some clinical improvement and were very well tolerated. Fluences ranging from 1 to 6 multiples of MEDs resulted in clearance of lesions in some patients with 6 MEDs producing clearance in 77% of patients. The number of treatments required to clear psoriatic lesions when 2 to 6 MEDs were used was 5.0 to 6.1 treatments. The only adverse events observed were erythema, which was asymptomatic in most subjects, and hyperpigmentation. CONCLUSIONS: Incoherent, targeted UV-B phototherapy is a safe and efficacious treatment modality for localized psoriasis. Its value in other UV-B responsive conditions should be further investigated. PMID- 16365256 TI - Dose-response and time-course characteristics of UV-A1 erythema. AB - OBJECTIVE: To determine the time course and dose-response characteristics of UV A1 erythema in the Tayside region of Scotland. DESIGN: Adult volunteers (skin types I and II [n = 13] and III and IV [n = 11]) were exposed to geometric dose series of UV-A1 irradiation from a high-output source on photoprotected lower back and inner forearm skin. SETTING: Photobiology unit in a university hospital. MAIN OUTCOME MEASURES: The minimal erythema dose (MED) was recorded visually and erythema was assessed objectively by erythema meter at 4, 8, 24, and 48 hours after exposure. RESULTS: Peak erythema (lowest visual MED) was seen at 8 hours on the back and arm in 11 subjects with skin types I and II and on the back at 8 hours in 9 subjects and on the arm at 4 hours in 10 subjects with skin types III and IV. The lowest median (range) MED was 20 J/cm(2) (14-56 J/cm(2)) on the back and 42 J/cm(2) (20 to >80 J/cm(2)) on the arm at 8 hours for subjects with skin types I and II and 28 J/cm(2) (20-112 J/cm(2)) at 8 hours on the back and 56 J/cm(2) (28-80 J/cm(2)) at 4 hours on the arm for subjects with skin types III and IV. The D(0.025), an objective measure that corresponds approximately to the visual MED, demonstrated a broad peak from 8 to 24 hours. CONCLUSIONS: Our local population is more erythemally sensitive to UV-A1 radiation than reports suggest. Daily dose regimens may risk cumulative erythema. Lower starting doses should be used in this population. The wide range of MEDs highlights the need for MED testing. PMID- 16365257 TI - Soak and smear: a standard technique revisited. AB - BACKGROUND: Atopic dermatitis, nummular eczema, chronic hand dermatitis, palmar plantar psoriasis, and xerotic eczema are common inflammatory skin conditions. They may be refractory to conventional topical and even systemic treatment. Little evidence is available that demonstrates the benefits of aggressive topical treatment of patients with these disorders. OBJECTIVE: To describe a simple, inexpensive, effective topical treatment with an accompanying patient educational sheet. DESIGN: A retrospective study of 28 patients referred to a tertiary care center for refractory chronic pruritic eruptions. Intervention with a plain water 20-minute soak followed by smearing of mid-strength to high-strength corticosteroid ointment led to clearing or dramatic improvement. RESULTS: Objective and symptomatic improvement was obtained from aggressive topical treatment. It was well accepted in this group of referral patients. CONCLUSIONS: Hydration for 20 minutes before bedtime followed by ointment application to wet skin and alteration of cleansing habits is an effective method for caring for several common skin conditions. Prospective studies are needed to further validate these findings. PMID- 16365258 TI - Skin excision and osteophyte removal is not required in the surgical treatment of digital myxoid cysts. AB - BACKGROUND: Digital myxoid cysts (DMCs) are ganglia of the adjacent distal interphalangeal joint (DIPJ) caused by leakage of fluid from the joint into the surrounding tissues. The connection between the DIPJ and the cyst can be identified by the injection of methylene blue into the DIPJ. However, the injection of methylene blue into the DIPJ is difficult and time-consuming. Based on this understanding of the cause of DMCs, we have used a surgical technique to treat DMCs without the need for skin excision. Herein, we have adapted the technique and demonstrated that precise leakage point identification is not required for treatment success, thus reducing the potential postoperative morbidity, reducing the operative time, and simplifying the surgical technique. DESIGN: This was a prospective, open, nonrandomized trial of therapy. A skin flap was designed to include the cyst and tissues from the cyst to the DIPJ. No skin excision was required, and no osteophyte removal was attempted. SETTING: University dermatology department. PATIENTS: Thirty-two consecutive symptomatic subjects with 26 finger DMCs and 6 toe DMCs. No patient had been previously treated. MAIN OUTCOME MEASURES: Clinical assessment postoperatively and recurrence rate after a minimum follow-up of 8 months. RESULTS: Of the 26 finger DMCs, 24 (92.3%) remained healed at 8 months; and of the 6 toe DMCs, 2 (33.3%) remained healed at 8 months. CONCLUSIONS: Digital myxoid cysts are caused by leakage of joint fluid from the DIPJ to the cyst. The leakage point is sealed in the healing process that occurs after a flap is raised and re-sited. The flap must be designed to include the undersurface of the cyst and the tissues between the DIPJ and the cyst. No skin excision or osteophyte removal is required. The procedure is not recommended for DMCs of the toes. PMID- 16365259 TI - Two novel TP63 mutations associated with the ankyloblepharon, ectodermal defects, and cleft lip and palate syndrome: a skin fragility phenotype. AB - BACKGROUND: Ankyloblepharon, ectodermal defects, and cleft lip and palate (AEC) syndrome is a rare autosomal dominant disorder caused by mutations in the sterile alpha motif region of TP63, a homologue of the tumor suppressor TP53. Recent structure-function studies have identified complexities in the genotype-phenotype correlation of the p63 syndromes. OBSERVATIONS: We report 2 sporadic cases of AEC syndrome in infants. Both patients demonstrated skin erosions with prominent scalp involvement. Histologic studies demonstrated mild basal layer vacuolization and rare dyskeratotic keratinocytes, with evidence of both acantholysis and cytolysis at the blister edge. Immunohistochemistry using anti-p63 monoclonal antibody demonstrated basal epidermal nuclear staining in both healthy control and patient tissue samples. Ultrastructural studies showed focal disruption of anchoring fibrils near the blister edge of one patient and normal desmosomes, hemidesmosomes, and basement membrane zone in the nonblistered skin of the other patient. The DNA analysis of each patient revealed 2 novel missense mutations in the TP63 gene that resulted in L514S and R555P amino acid substitutions within the sterile alpha motif region of the p63 protein. CONCLUSIONS: We report 2 novel TP63 mutations resulting in AEC syndrome. The R555P mutation is the most carboxy terminal of all the reported AEC missense mutations of p63. The presence of skin fragility, manifested as erosive skin lesions in body areas in addition to the scalp, is postulated to be an important diagnostic feature of AEC syndrome. PMID- 16365260 TI - Two brothers with mild congenital erythropoietic porphyria due to a novel genotype. AB - BACKGROUND: Congenital erythropoietic porphyria (CEP) is a rare autosomal recessive disease caused by the deficient activity of the heme biosynthetic enzyme, uroporphyrinogen III synthase (URO-synthase), and the accumulation of the nonphysiologic and phototoxic porphyrin I isomers. Clinical manifestations range from severe mutilation to mild erosions and blisters on sun-exposed areas. Evaluation of the URO-synthase mutation and residual enzyme activity has been correlated with the phenotypic expression of the disease. OBSERVATIONS: We describe 16- and 4-year-old brothers with CEP with a mild phenotype due to a novel genotype, one allele having a promoter mutation (-76G-->A) and the other having an exonic missense mutation (G225S). The father and a 4-year-old fraternal twin brother were carriers of the -76G-->A mutation, whereas the mother and a 15 year-old brother were carriers of the G225S mutation. Previous in vitro expression studies demonstrated that the G225S mutation severely decreased URO synthase activity to 1.2% of normal, whereas the promoter mutation decreased the activity to approximately 50% of wild type, accounting for the mild clinical phenotype. CONCLUSION: The mild disease phenotype in these patients is a further example of the clinical heterogeneity seen in CEP and is additional proof that in vitro enzyme expression studies provide dependable genotype-phenotype correlations. PMID- 16365261 TI - Relationship between smoking and the clinical severity of psoriasis. AB - OBJECTIVE: To evaluate the association between different components of smoking history and the clinical severity of psoriasis. DESIGN: A hospital-based cross sectional study. SETTING: Inpatient wards of a hospital for skin diseases in Rome, Italy. PATIENTS: A total of 818 adults with psoriasis. MAIN OUTCOME MEASURE: The Psoriasis Area and Severity Index was used to assess the clinical severity of psoriasis between February 21, 2000, and February 19, 2002. RESULTS: After adjustment for potential confounders (sex, age, body mass index, psychological distress, family history of psoriasis, duration of psoriasis disease, and alcohol consumption), high intensity of smoking (>20 cigarettes daily) vs a lower level of consumption (< or =10 cigarettes daily) was associated with a more than 2-fold increased risk of clinically more severe psoriasis (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.2-4.1). Cigarette-years, measured as the product of the intensity and duration (years) of smoking, significantly increased the risk of clinically more severe psoriasis after adjustment for confounding factors (OR,1.3; 95% CI, 1.0-1.6, for a 600-U increase in cigarette-years). Separate analyses for men and women showed that the effect of cigarette-years was stronger for women (OR, 1.8; 95% CI, 1.2-2.6, for a 400-U increase in cigarette-years) than for men (OR, 1.2; 95% CI, 0.9-1.6, for a 700-U increase in cigarette-years). CONCLUSION: Smoking is associated with the clinical severity of psoriasis and highlights the importance of smoking cessation in patients with psoriasis. PMID- 16365262 TI - Another paradigm lost or just a paradox? PMID- 16365263 TI - Advances in psoriasis. PMID- 16365264 TI - Skin erosions and wound healing in ankyloblepharon-ectodermal defect-cleft lip and/or palate. PMID- 16365265 TI - Multiple firm, painless erythematous papules with a yellowish hue. PMID- 16365266 TI - Bluish nodules on the trunk and arms. PMID- 16365267 TI - Bluish papule in a middle-aged man. PMID- 16365268 TI - Lax skin masses in a 47-year-old woman.. PMID- 16365269 TI - The skinny on psoriasis and obesity. PMID- 16365270 TI - AMEVIVE's advertisements: problematic? PMID- 16365271 TI - Successful treatment of plaque psoriasis with self-administered subcutaneous alefacept. PMID- 16365272 TI - A consideration of statistical association. PMID- 16365273 TI - Transmission of parthenium dermatitis by clothing. PMID- 16365274 TI - Intralesional immunotherapy for genital warts. PMID- 16365275 TI - Infliximab combined with methotrexate as long-term treatment for erythrodermic psoriasis. PMID- 16365277 TI - Dermoscopy of molluscum contagiosum. PMID- 16365278 TI - The 73-kDa heat shock cognate protein is a CXCR4 binding protein that regulates the receptor endocytosis and the receptor-mediated chemotaxis. AB - The CXCR4 chemokine receptor is a G protein-coupled receptor that plays an important role in leukocyte homing, cancer metastasis, and human immunodeficiency virus infection. In response to ligand stimulation, chemokine receptors undergo endocytosis through clathrin-coated vesicle (CCV). Uncoating of CCV, a process involving heat shock cognate protein and several other proteins, is critical for fusion of CCV to endosomal compartments. The present study demonstrated that CXCR4 was associated with the 73-kDa heat shock cognate protein (Hsc73) in human embryonic kidney 293 cells in response to ligand stimulation. Truncation of the carboxyl terminal domain of CXCR4 reduced the association with Hsc73 and a glutathione S-transferase-CXCR4 carboxyl terminal fusion protein associated with Hsc73 in vitro, suggesting involvement of the carboxyl terminal domain of the receptor in the interaction. In response to ligand stimulation, CXCR4 underwent internalization and colocalization with Hsc73, but the receptor endocytosis was blocked by knockdown of Hsc73 with RNA interference. Moreover, Hsc73 knockdown significantly reduced the CXCR4-mediated chemotaxis of U87 glioma cell lines. These findings suggest that Hsc73 plays a role in chemokine receptor trafficking and the receptor-mediated chemotaxis. PMID- 16365279 TI - Activation of dopamine D4 receptors induces synaptic translocation of Ca2+/calmodulin-dependent protein kinase II in cultured prefrontal cortical neurons. AB - One of the important targets of dopamine D4 receptors in prefrontal cortex (PFC) is the multifunctional Ca2+/calmodulin-dependent protein kinase II (CaMKII). In the present study, we investigated the effect of D4 receptor activation on subcellular localization of CaMKII. We found that activation of D4 receptors, but not D2 receptors, induced a rapid translocation of alpha-CaMKII from cytosol to postsynaptic sites in cultured PFC neurons. Activated CaMKII (Thr286 phospho CaMKII) was also redistributed to postsynaptic sites after D4 receptor stimulation. The translocation was blocked by inhibiting the phospholipase C/inositol 1,4,5-trisphosphate receptor/Ca2+ signaling. Point mutation of the calmodulin binding site (Ala302), but not the autophosphorylation site (Thr286), of alpha-CaMKII prevented the D4-induced CaMKII translocation. Moreover, D4 receptors failed to induce CaMKII translocation in the presence of an actin stabilizer, and D4 activation reduced the binding of CaMKII to F-actin. Concomitant with the synaptic accumulation of alpha-CaMKII in response to D4 receptor activation, a D4-induced increase in the CaMKII phosphorylation of alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor glutamate receptor 1 (GluR1) subunits and the amplitude of AMPA receptor-mediated excitatory postsynaptic currents was also observed. Thus, our results show that D4 receptor activation induces the synaptic translocation of CaMKII through a mechanism involving Ca2+/calmodulin and F-actin, which facilitates the regulation of synaptic targets of CaMKII, such as AMPA receptors. PMID- 16365280 TI - Ester derivatives of tournefolic acid B attenuate N-methyl-D-aspartate-mediated excitotoxicity in rat cortical neurons. AB - The effects of tournefolic acid B (TAB) and two ester derivatives, TAB methyl ester (TABM) and TAB ethyl ester (TABE), on N-methyl-D-aspartate (NMDA)-mediated excitotoxicity and the underlying mechanisms were investigated. Treatment with 50 microM NMDA elicited neuronal death by 48.7 +/- 5.1%, coinciding with the appearance of injured morphology. TABM (50 microM) attenuated the NMDA-induced cell death by 60.9 +/- 19.7%, and to a lesser extent by TABE. The NMDA-mediated activation of calpain was not affected by TABM and TABE, as determined by the cleavage of alpha-spectrin. NMDA increased the activity of caspases 2, 3, 6, 8, and 9 and reached the maximum after 8-h treatment. TABM and TABE abrogated NMDA induced activation of caspases 2, 3, 6, and 8 by approximately 80 to 90% and 50 to 60%, respectively, and to a higher extent for caspase 9. TABM and TABE also blocked the NMDA-mediated activation of caspase 12. Furthermore, TABM and TABE eliminated the NMDA-induced accumulation of superoxide anion (O2-*). NMDA evoked significant depolarization of mitochondria, whereas TABM elicited a mild decrease of mitochondrial membrane potential as determined by tetramethylrhodamine methyl ester perchlorate. NMDA treatment induced elevation of Ca2+ levels in cytosol, endoplasmic reticulum (ER), and mitochondria. TABM (50 microM) significantly diminished the NMDA-induced elevation of Ca2+ levels in mitochondria and ER but not cytosol. Therefore, TABM decreased mitochondrial membrane potential and attenuated the NMDA-mediated Ca2+-loading in ER and mitochondria. These events subsequently eliminated the accumulation of O2-* and blocked the activation of caspase cascade, thereby conferring their neuroprotective effects on NMDA mediated excitotoxicity. PMID- 16365281 TI - Opposing functions of spinal M2, M3, and M4 receptor subtypes in regulation of GABAergic inputs to dorsal horn neurons revealed by muscarinic receptor knockout mice. AB - Spinal muscarinic acetylcholine receptors (mAChRs) play an important role in the regulation of nociception. To determine the role of individual mAChR subtypes in control of synaptic GABA release, spontaneous inhibitory postsynaptic currents (sIPSCs) and miniature IPSCs (mIPSCs) were recorded in lamina II neurons using whole-cell recordings in spinal cord slices of wild-type and mAChR subtype knockout (KO) mice. The mAChR agonist oxotremorine-M (3-10 microM) dose dependently decreased the frequency of GABAergic sIPSCs and mIPSCs in wild-type mice. However, in the presence of the M2 and M4 subtype-preferring antagonist himbacine, oxotremorine-M caused a large increase in the sIPSC frequency. In M3 KO and M1/M3 double-KO mice, oxotremorine-M produced a consistent decrease in the frequency of sIPSCs, and this effect was abolished by himbacine. We were surprised to find that in M2/M4 double-KO mice, oxotremorine-M consistently increased the frequency of sIPSCs and mIPSCs in all neurons tested, and this effect was completely abolished by 4-diphenylacetoxy-N-methylpiperidine methiodide, an M3 subtype-preferring antagonist. In M2 or M4 single-KO mice, oxotremorine-M produced a variable effect on sIPSCs; it increased the frequency of sIPSCs in some cells but decreased the sIPSC frequency in other neurons. Taken together, these data strongly suggest that activation of the M3 subtype increases synaptic GABA release in the spinal dorsal horn of mice. In contrast, stimulation of presynaptic M2 and M4 subtypes predominantly attenuates GABAergic inputs to dorsal horn neurons in mice, an action that is opposite to the role of M2 and M4 subtypes in the spinal cord of rats. PMID- 16365282 TI - Loss of synaptic D1 dopamine/N-methyl-D-aspartate glutamate receptor complexes in L-DOPA-induced dyskinesia in the rat. AB - Glutamate-mediated mechanisms are related to the motor complications of L-DOPA therapy in Parkinson's disease (PD). In striatal postsynaptic densities (PSD), the dopamine D1 receptor (D1R) is part of an oligomeric complex with the glutamate N-methyl-D-aspartate receptor (NMDAR), determining the strength of corticostriatal transmission. We studied D1R/NMDAR complex alterations induced by L-DOPA in the 6-hydroxydopamine-lesioned rat model of PD. L-DOPA-treated hemiparkinsonian rats were determined to be dyskinetic or nondyskinetic based on behavioral testing. D1R/NMDAR assemblies containing NR1-C2 and NR2B subunits were decreased in the PSD of lesioned striatum. Short-term L-DOPA administration improved akinesia and restored the synaptic abundance of D1R, NR1-C2 and NR2B. Prolonged L-DOPA treatment also normalized synaptic D1R/NMDAR complexes in nondyskinetic rats, but remarkably reduced them in the dyskinetic group without changing their interaction. This decrease involved NR1-C2, NR1-C2', NR2A, and NR2B subunits. The composition of residual synaptic D1R/NMDAR complexes in dyskinetic rats may thus be different from that observed in lesioned rats, suggesting that expression of different motor dysfunctions might be related to the receptor profile at corticostriatal synapses. The levels of D1R/NMDAR complexes were unchanged in total striatal membrane proteins, suggesting that the decrease of these species in the PSD is likely to reflect an altered receptor trafficking. In human embryonic kidney 293 cells expressing the D1R/NMDAR, complex costimulation of both D1R and NMDAR, but not individual receptor activation, promoted internalization, suggesting that development of dyskinesias might be related to agonist-mediated down-regulation of the D1R/NMDAR complex at corticostriatal synapses. PMID- 16365283 TI - The mitochondrial theory of aging and its relationship to reactive oxygen species damage and somatic mtDNA mutations. PMID- 16365284 TI - The growth of business firms: theoretical framework and empirical evidence. AB - We introduce a model of proportional growth to explain the distribution P(g)(g) of business-firm growth rates. The model predicts that P(g)(g) is exponential in the central part and depicts an asymptotic power-law behavior in the tails with an exponent zeta = 3. Because of data limitations, previous studies in this field have been focusing exclusively on the Laplace shape of the body of the distribution. In this article, we test the model at different levels of aggregation in the economy, from products to firms to countries, and we find that the predictions of the model agree with empirical growth distributions and size variance relationships. PMID- 16365285 TI - Adoptive transfer of T cells modified with a humanized chimeric receptor gene inhibits growth of Lewis-Y-expressing tumors in mice. AB - In this study, human T cells were provided with a reactivity against the Lewis-Y (Le(Y)) carbohydrate antigen, which is overexpressed on 70% of epithelial-derived tumors, but not normally recognized by T cells. Antitumor reactivity was achieved by transduction of T cells with a gene encoding a cell-surface chimeric receptor composed of single-chain anti-Le(Y) antibody linked to an enhanced cytoplasmic signaling domain made up of CD28 and CD3-zeta. Importantly, the single-chain antibody was humanized to try to reduce potential problems of human anti-mouse antibody responses in patients receiving chimeric receptor-modified T cells in future clinical trials. T cells expressing the chimeric receptor were demonstrated to secrete cytokines and proliferate in response to receptor ligation and lysed Le(Y+) tumors in vitro. Another aspect of this study was the finding that no activity was observed against normal tissue, as represented by autologous neutrophils that express low levels of Le(Y). Significantly, systemic delivery of anti-Le(Y) T cells dramatically inhibited established s.c. human ovarian OVCAR-3 tumors (a recognized difficult model to treat) in mice. Finally, we demonstrated that anti-Le(Y) T cells preferentially expanded or accumulated in the tumor compared with control empty vector T cells, thereby providing mechanistic insight into the specific antitumor response. This study supports the use of humanized gene-modified T cells as a potential therapy for Le(Y+) malignancies. PMID- 16365286 TI - Indigestion is a plant's best defense. PMID- 16365287 TI - The identification and characterization of two phosphatidylinositol-4,5 bisphosphate 4-phosphatases. AB - Numerous inositol polyphosphate 5-phosphatases catalyze the degradation of phosphatidylinositol-4,5-bisphosphate (PtdIns-4,5-P(2)) to phosphatidylinositol-4 phosphate (PtdIns-4-P). An alternative pathway to degrade PtdIns-4,5-P(2) is the hydrolysis of PtdIns-4,5-P(2) by a 4-phosphatase, leading to the production of PtdIns-5-P. Whereas the bacterial IpgD enzyme is known to catalyze this reaction, no such mammalian enzyme has been found. We have identified and characterized two previously undescribed human enzymes, PtdIns-4,5-P(2) 4-phosphatase type I and type II, which catalyze the hydrolysis of PtdIns-4,5-P(2) to phosphatidylinositol 5-phosphate (PtdIns-5-P). Both enzymes are ubiquitously expressed and localize to late endosomal/lysosomal membranes in epithelial cells. Overexpression of either enzyme in HeLa cells increases EGF-receptor degradation upon EGF stimulation. PMID- 16365288 TI - Expression of a homodimeric type I cytokine receptor is required for JAK2V617F mediated transformation. AB - A recurrent somatic activating mutation in the nonreceptor tyrosine kinase JAK2 (JAK2V617F) occurs in the majority of patients with the myeloproliferative disorders polycythemia vera, essential thrombocythemia, myelofibrosis with myeloid metaplasia, and, less commonly, chronic myelomonocytic leukemia. We do not understand the basis for the specificity of the JAK2V617F mutation in clonal disorders of the myeloid, but not lymphoid, lineage, nor has the basis for the pleiotropic phenotype of JAK2V617F-associated myeloproliferative disorders been delineated. However, the presence of the identical mutation in patients with related, but clinicopathologically distinct, myeloid disorders suggests that interactions between the JAK2V617F kinase and other signaling molecules may influence the phenotype of hematopoietic progenitors expressing JAK2V617F. Here, we show that coexpression of the JAK2V617F mutant kinase with a homodimeric Type I cytokine receptor, the erythropoietin receptor (EpoR), the thrombopoietin receptor, or the granulocyte colony-stimulating-factor receptor, is necessary for transformation of hematopoietic cells to growth-factor independence and for hormone-independent activation of JAK-STAT signaling. Furthermore, EpoR mutations that impair erythropoietin-mediated JAK2 or STAT5 activation also impair transformation mediated by the JAK2V617F kinase, indicating that JAK2V617F requires a cytokine receptor scaffold for its transforming and signaling activities. Our results reveal the molecular basis for the prevalence of JAK2V617F in diseases of myeloid lineage cells that express these Type I cytokine receptors but not in lymphoid lineage cells that do not. PMID- 16365289 TI - A MAPK gene from Dead Sea fungus confers stress tolerance to lithium salt and freezing-thawing: Prospects for saline agriculture. AB - The Dead Sea is one of the most saline lakes on earth ( approximately 340 g/liter salinity) and is approximately 10 times saltier than the oceans. Eurotium herbariorum, a common fungal species, was isolated from its water. EhHOG gene, encoding a mitogen-activated protein kinase (MAPK) that plays an essential role in the osmoregulatory pathway in yeast and many other eukaryotes, was isolated from E. herbariorum. The deduced amino acid sequences of EhHOG indicated high similarity with homologous genes from Aspergillus nidulans, Saccharomyces cerevisiae, and Schizosaccharomyces pombe and contained a TGY motif for phosphorylation by MAPK kinase. When EhHOG was expressed in S. cerevisiae hog1Delta mutant, the growth and aberrant morphology of hog1Delta mutant was restored under high osmotic stress condition. Moreover, intracellular glycerol content in the transformant increased to a much higher level than that in the mutant during salt-stress situations. hog1Delta mutant complemented by EhHOG outperformed the wild type or had higher genetic fitness under high Li(+) and freezing-thawing conditions. The present study revealed the putative presence of a high-osmolarity glycerol response (HOG) pathway in E. herbariorum and the significance of EhHOG in osmotic regulation, heat stress, freeze stress, and oxidative stress. The Dead Sea is becoming increasingly more saline while the fungi living in it evolutionarily adapt to its high-saline environment, particularly with the extraordinarily high Li(+) concentration. The Dead Sea is potentially an excellent model for studies of evolution under extreme environments and is an important gene pool for future agricultural genetic engineering prospects. PMID- 16365290 TI - A 200-kb region of human chromosome 22q11.2 confers antipsychotic-responsive behavioral abnormalities in mice. AB - Human chromosome 22q11.2 has been implicated in various behavioral abnormalities, including schizophrenia and other neuropsychiatric/behavioral disorders. However, the specific genes within 22q11.2 that contribute to these disorders are still poorly understood. Here, we show that an approximately 200-kb segment of human 22q11.2 causes specific behavioral abnormalities in mice. Mice that overexpress an approximately 200-kb region of human 22q11.2, containing CDCrel, GP1Bbeta, TBX1, and WDR14, exhibited spontaneous sensitization of hyperactivity and a lack of habituation. These effects were ameliorated by antipsychotic drugs. The transgenic mice were also impaired in nesting behavior. Although Tbx1 has been shown to be responsible for many physical defects associated with 22q11.2 haploinsufficiency, Tbx1 heterozygous mice did not display these behavioral abnormalities. Our results show that the approximately 200-kb region of 22q11.2 contains a gene(s) responsible for behavioral abnormalities and suggest that distinct genetic components within 22q11.2 mediate physical and behavioral abnormalities. PMID- 16365291 TI - The role of microRNA genes in papillary thyroid carcinoma. AB - Apart from alterations in the RET/PTC-RAS-BRAF pathway, comparatively little is known about the genetics of papillary thyroid carcinoma (PTC). We show that numerous microRNAs (miRNAs) are transcriptionally up-regulated in PTC tumors compared with unaffected thyroid tissue. A set of five miRNAs, including the three most up-regulated ones (miR-221, -222, and -146), distinguished unequivocally between PTC and normal thyroid. Additionally, miR-221 was up regulated in unaffected thyroid tissue in several PTC patients, presumably an early event in carcinogenesis. Tumors in which the up-regulation (11- to 19-fold) of miR-221, -222, and -146 was strongest showed dramatic loss of KIT transcript and Kit protein. In 5 of 10 such cases, this down expression was associated with germline single-nucleotide changes in the two recognition sequences in KIT for these miRNAs. We conclude that up-regulation of several miRs and regulation of KIT are involved in PTC pathogenesis, and that sequence changes in genes targeted by miRNAs can contribute to their regulation. PMID- 16365292 TI - Perinatal anoxia degrades auditory system function in rats. AB - Little is known about the neural bases of the reduced auditory and cortical processing speeds that have been recorded in language-impaired, autistic, schizophrenic, and other disabled human populations. Although there is strong evidence for genetic contributions to etiologies, epigenetic factors such as perinatal anoxia (PA) have been argued to be contributors, or causal, in a significant proportion of cases. In this article, we explored the consequences of PA on this elementary aspect of auditory behavior and on auditory system function in rats that were briefly perinatally anoxic. PA rats had increased acoustic thresholds and reduced processing efficiencies recorded in an auditory behavioral task. These rats had modestly increased interpeak intervals in their auditory brainstem responses, and substantially longer latencies in poststimulus time histogram responses recorded in the primary auditory cortex. The latter were associated with degraded primary auditory cortex receptive fields and a disrupted tonotopy. These processing deficits are consistent with the parallel behavioral and physiological deficits recorded in children and adults with a history of language-learning impairment and autism. PMID- 16365293 TI - The immunomodulator glatiramer acetate augments the expression of neurotrophic factors in brains of experimental autoimmune encephalomyelitis mice. AB - Neurotrophins (NTs) such as BDNF, NT-3, and NT-4 are important modulators of neuronal function and survival. Their expression in the CNS after various insults is thus of major therapeutic consequence. Glatiramer acetate [(GA) Copaxone], an approved drug for the treatment of multiple sclerosis, has been shown to induce Th2/3 cells that accumulate in the CNS, expressing in situ antiinflammatory cytokines and BDNF. In the present study, we investigated whether s.c. injections of GA, applied at various stages of experimental autoimmune encephalomyelitis, affect the expression of NTs, particularly BDNF, in the brain. In untreated experimental autoimmune encephalomyelitis mice, the expression of NTs was elevated shortly after disease appearance but subsequently declined below that of naive mice. In contrast, GA treatment led to sustained augmentation in the expression of BDNF, NT-3, and NT-4 in various brain regions as demonstrated by histological analysis of immunostained brain sections. GA treatment, even when started 45 days after disease induction, restored the impaired level of NTs to that of healthy mice. BDNF elevation after GA treatment was demonstrated on both protein and mRNA levels. Prominent staining was manifested not only by infiltrating GA-induced T cells, but also by CNS resident cells (neurons and astrocytes), indicative of a bystander therapeutic effect. Of importance, in GA treated mice, intense BDNF expression was manifested by neuronal progenitors that migrated into lesions in injured regions. These results indicate that the immunomodulator GA exerts not only an antiinflammatory effect, but also enhances neuroprotection and regeneration of neural elements in the diseased brain. PMID- 16365294 TI - High-dimensional and large-scale phenotyping of yeast mutants. AB - One of the most powerful techniques for attributing functions to genes in uni- and multicellular organisms is comprehensive analysis of mutant traits. In this study, systematic and quantitative analyses of mutant traits are achieved in the budding yeast Saccharomyces cerevisiae by investigating morphological phenotypes. Analysis of fluorescent microscopic images of triple-stained cells makes it possible to treat morphological variations as quantitative traits. Deletion of nearly half of the yeast genes not essential for growth affects these morphological traits. Similar morphological phenotypes are caused by deletions of functionally related genes, enabling a functional assignment of a locus to a specific cellular pathway. The high-dimensional phenotypic analysis of defined yeast mutant strains provides another step toward attributing gene function to all of the genes in the yeast genome. PMID- 16365295 TI - Mechanistic insight into the allosteric activation of a ubiquitin-conjugating enzyme by RING-type ubiquitin ligases. AB - Ubiquitin-conjugating enzymes (E2s) collaborate with the ubiquitin-activating enzyme (E1) and ubiquitin ligases (E3s) to attach ubiquitin to target proteins. RING-containing E3s simultaneously bind to E2s and substrates, bringing them into close proximity and thus facilitating ubiquitination. We show herein that, although the E3-binding site on the human E2 UbcH5b is distant from its active site, two RING-type minimal E3 modules lacking substrate-binding functions greatly stimulate the rate of ubiquitin release from the UbcH5b-ubiquitin thioester. Using statistical coupling analysis and mutagenesis, we identify and characterize clusters of coevolving and functionally linked residues within UbcH5b that span its E3-binding and active sites. Several UbcH5b mutants are defective in their stimulation by E3s despite their abilities to bind to these E3s, to form ubiquitin thioesters, and to release ubiquitin at a basal rate. One such mutation, I37A, is distant from both the active site and the E3-binding site of UbcH5b. Our studies reveal structural determinants for communication between distal functional sites of E2s and suggest that RING-type E3s activate E2s allosterically. PMID- 16365296 TI - Integrity matters: linking nuclear architecture to lifespan. PMID- 16365297 TI - A dynamic switch between inhibitory and excitatory currents in a neuronal glutamate transporter. AB - Excitatory amino acid transporters (EAATs) terminate glutamatergic synaptic transmission and maintain extracellular glutamate concentrations in the central nervous system below excitotoxic levels. In addition to sustaining a secondary active glutamate transport, EAAT glutamate transporters also function as anion selective channels. Here, we report a gating process that makes anion channels associated with a neuronal glutamate transporter, EAAT4, permeable to cations and causes a selective increase of the open probability at voltages negative to the actual current reversal potential. The activation process depends on both membrane potential and extracellular glutamate concentration and causes an accumulation of EAAT4 anion channels in a state favoring cation influx and anion efflux. Gating of EAAT4 anion channels thus allows a switch between inhibitory currents in resting cells and excitatory currents in electrically active cells. This transporter-mediated conductance could modify the excitability of Purkinje neurons, providing them with an unprecedented mechanism for adaptation. PMID- 16365298 TI - Complex I deficiency primes Bax-dependent neuronal apoptosis through mitochondrial oxidative damage. AB - Dysfunction of mitochondrial complex I is a feature of human neurodegenerative diseases such as Leber hereditary optic neuropathy and Parkinson's disease. This mitochondrial defect is associated with a recruitment of the mitochondrial dependent apoptotic pathway in vivo. However, in isolated brain mitochondria, complex I dysfunction caused by either pharmacological or genetic means fails to directly activate this cell death pathway. Instead, deficits of complex I stimulate intramitochondrial oxidative stress, which, in turn, increase the releasable soluble pool of cytochrome c within the mitochondrial intermembrane space. Upon mitochondrial permeabilization by the cell death agonist Bax, more cytochrome c is released to the cytosol from brain mitochondria with impaired complex I activity. Given these results, we propose a model in which defects of complex I lower the threshold for activation of mitochondrial-dependent apoptosis by Bax, thereby rendering compromised neurons more prone to degenerate. This molecular scenario may have far-reaching implications for the development of effective neuroprotective therapies for these incurable illnesses. PMID- 16365299 TI - Deregulation of common genes by c-Myc and its direct target, MT-MC1. AB - In addition to its role in cancer, the c-Myc oncoprotein controls many normal cellular processes as a consequence of its function as a basic helix-loop-helix leucine zipper transcription factor. Determining which of the myriad genes under c-Myc control are relevant for these various roles is thus a major challenge. mt mc1 is a direct c-Myc target gene whose overexpression recapitulates multiple c Myc phenotypes, including transformation. Using transcriptional profiling, we now show that MT-MC1-overexpressing myeloid cells misregulate a total of 47 distinct transcripts, a large proportion of which are involved in signal transduction and/or cancer. Analysis of these genes reveals a consensus promoter structure consisting of multiple, often closely spaced c-Myc binding sites and three additional Wilm's tumor and Egr1-like motifs. More than one-third of MT-MC1 target genes are also clustered on six cancer-associated chromosomal loci. Most surprisingly, all of the transcripts examined also are regulated by c-Myc. Finally, an estrogen receptor-MT-MC1 fusion protein was used to establish that all examined transcripts were regulated directly by the chimeric protein. Our results thus indicate that MT-MC1 target genes largely comprise a subset of those regulated by c-Myc. We propose that the properties imparted by MT-MC1 are the result of its control of a small and select c-Myc target gene population. PMID- 16365300 TI - Femtochemistry of orange II in solution and in chemical and biological nanocavities. AB - In this work, we report on studies of the nature of the dynamics and hydrophobic binding in cyclodextrins and human serum albumin protein complexes with orange II. With femtosecond time resolution, we examined the proton-transfer and trans cis isomerization reactions of the ligand in these nanocavities and in pure solvents. Because of confinement at the ground state, the orientational motion in the formed phototautomer is restricted, leading to a rich dynamics. Therefore, the emission lifetimes span a large window of tens to hundreds of picoseconds in the cavities. Possible H-bond interactions between the guest and cyclodextrin do not affect the caged dynamics. For the protein-ligand complexes, slow diffusional motion ( approximately 630 ps) observed in the anisotropy decay indicates that the binding structure is not completely rigid, and the embedded guest is not frozen with the hydrophobic pocket. The ultrafast isomerization and decays are explained in terms of coupling motions between N-N and C-N stretching modes of the formed tautomer. We discuss the role of confinement on the trans-cis isomerization with the cavities and its relationships to frequency and time domains of nanostructure emission. PMID- 16365301 TI - Extreme conformational diversity in human telomeric DNA. AB - DNA with tandem repeats of guanines folds into G-quadruplexes made of a stack of G-quartets. In vitro, G-quadruplex formation inhibits telomere extension, and POT1 binding to the single-stranded telomeric DNA enhances telomerase activity by disrupting the G-quadruplex structure, highlighting the potential importance of the G-quadruplex structure in regulating telomere length in vivo. We have used single-molecule spectroscopy to probe the dynamics of human telomeric DNA. Three conformations were observed in potassium solution, one unfolded and two folded, and each conformation could be further divided into two species, long-lived and short-lived, based on lifetimes of minutes vs. seconds. Vesicle encapsulation studies suggest that the total of six states detected here is intrinsic to the DNA. Folding was severely hindered by replacing a single guanine, showing only the shortlived species. The long-lived folded states are dominant in physiologically relevant conditions and probably correspond to the parallel and antiparallel G-quadruplexes seen in high-resolution structural studies. Although rare under these conditions, the short-lived species determine the overall dynamics because they bridge the different long-lived species. We propose that these previously unobserved transient states represent the early and late intermediates toward the formation of stable G-quadruplexes. The major compaction occurs between the early and late intermediates, and it is possible that local rearrangements are sufficient in locking the late intermediates into the stably folded forms. The extremely diverse conformations of the human telomeric DNA may have mechanistic implications for the proteins and drugs that recognize G-rich sequences. PMID- 16365302 TI - Epidermal transit of replication-arrested, undifferentiated keratinocytes in UV exposed XPC mice: an alternative to in situ apoptosis. AB - The interplay among nucleotide excision repair, cell-cycle regulation, and apoptosis in the UV-exposed epidermis is extremely important to avoid mutations and malignant transformation. In Xpc(-/-) mice deficient in global genome nucleotide excision repair (GGR), a cell-cycle arrest of epidermal cells in late S-phase [with near-double normal diploid (4N) DNA content] was observed 48-72 h after UV exposure. This arrest resolved without apoptosis (96-168 h). We surmised that these arrested keratinocytes with persistent DNA damage were removed by epidermal turnover. In vivo BrdUrd pulse-chase labeling (>17 h after UV exposure) showed that DNA replication after UV exposure was resumed in Xpc(-/-) mice, but it did not reveal any evidence of retained BrdUrd-labeled S-phase cells in the basal layer of the epidermis at 72 h. Interestingly, by this time a maximum number of cytokeratin 10-negative and cytokeratin 5-positive cells had appeared in the suprabasal epidermal cell layers of UV-exposed Xpc(-/-) mice. Accumulation of these "basal cell"-like keratinocytes in the suprabasal layers was clearly aberrant and was not observed in WT and heterozygous mice. Flow cytometric analyses of single-cell suspensions from UV-exposed Xpc(-/-) epidermis further showed that the "near-4N" arrested cells retained cytokeratin 5 and lacked cytokeratin 10. Hence, we conclude that the arrested near-4N cells became detached from the basal layer without entering a proper differentiation program and were indeed subsequently lost through the epidermal turnover. This expulsion apparently constitutes an alternative route, different from in situ apoptosis, to eliminate DNA-damaged arrested cells from the epidermis. PMID- 16365303 TI - Brain estrogen deficiency accelerates Abeta plaque formation in an Alzheimer's disease animal model. AB - Much evidence indicates that women have a higher risk of developing Alzheimer's disease (AD) than do men. The reason for this gender difference is unclear. We hypothesize that estrogen deficiency in the brains of women with AD may be a key risk factor. In rapidly acquired postmortem brains from women with AD, we found greatly reduced estrogen levels compared with those from age- and gender-matched normal control subjects; AD and control subjects had comparably low levels of serum estrogen. We examined the onset and severity of AD pathology associated with estrogen depletion by using a gene-based approach, by crossing the estrogen synthesizing enzyme aromatase gene knockout mice with APP23 transgenic mice, a mouse model of AD, to produce estrogen-deficient APP23 mice. Compared with APP23 transgenic control mice, estrogen-deficient APP23 mice exhibited greatly reduced brain estrogen and early-onset and increased beta amyloid peptide (Abeta) deposition. These mice also exhibited increased Abeta production, and microglia cultures prepared from the brains of these mice were impaired in Abeta clearance/degradation. In contrast, ovariectomized APP23 mice exhibited plaque pathology similar to that observed in the APP23 transgenic control mice. Our results indicate that estrogen depletion in the brain may be a significant risk factor for developing AD neuropathology. PMID- 16365304 TI - Adaptation of nutrient supply to fetal demand in the mouse involves interaction between the Igf2 gene and placental transporter systems. AB - The mammalian fetus is unique in its dependence during gestation on the supply of maternal nutrients through the placenta. Maternal supply and fetal demand for nutrients need to be fine tuned for healthy growth and development of the fetus along its genetic trajectory. An altered balance between supply and demand can lead to deviations from this trajectory with long-term consequences for health. We have previously shown that in a knockout lacking the imprinted placental specific Igf2 transcript (P0), growth of the placenta is compromised from early gestation but fetal growth is normal until late gestation, suggesting functional adaptation of the placenta to meet the fetal demands. Here, we show that placental transport of glucose and amino acids are increased in the Igf2 P0(+/-) null and that this up-regulation of transport occurs, at least in part, through increased expression of the transporter genes Slc2a3 and Slc38a4, the imprinted member of the System A amino acid transporter gene family. Decreasing fetal demand genetically by removal of fetal Igf2 abolished up-regulation of both transport systems and reduced placental System A amino acid transport activity and expression of Slc38a2 in late gestation. Our results provide direct evidence that the placenta can respond to fetal demand signals through regulation of expression of specific placental transport systems. Thus, crosstalk between an imprinted growth demand gene (Igf2) and placental supply transporter genes (Slc38a4, Slc38a2, and Slc2a3) may be a component of the genetic control of nutrient supply and demand during mammalian development. PMID- 16365305 TI - Widespread genetic exchange among terrestrial bacteriophages. AB - Bacteriophages are the most numerous entities in the biosphere. Despite this numerical dominance, the genetic structure of bacteriophage populations is poorly understood. Here, we present a biogeography study involving 25 previously undescribed bacteriophages from the Cystoviridae clade, a group characterized by a dsRNA genome divided into three segments. Previous laboratory manipulation has shown that, when multiple Cystoviruses infect a single host cell, they undergo (i) rare intrasegment recombination events and (ii) frequent genetic reassortment between segments. Analyzing linkage disequilibrium (LD) within segments, we find no significant evidence of intrasegment recombination in wild populations, consistent with (i). An extensive analysis of LD between segments supports frequent reassortment, on a time scale similar to the genomic mutation rate. The absence of LD within this group of phages is consistent with expectations for a completely sexual population, despite the fact that some segments have >50% nucleotide divergence at 4-fold degenerate sites. This extraordinary rate of genetic exchange between highly unrelated individuals is unprecedented in any taxa. We discuss our results in light of the biological species concept applied to viruses. PMID- 16365306 TI - High-resolution protein folding with a transferable potential. AB - A generalized computational method for folding proteins with a fully transferable potential and geometrically realistic all-atom model is presented and tested on seven helix bundle proteins. The protocol, which includes graph-theoretical analysis of the ensemble of resulting folded conformations, was systematically applied and consistently produced structure predictions of approximately 3 A without any knowledge of the native state. To measure and understand the significance of the results, extensive control simulations were conducted. Graph theoretic analysis provides a means for systematically identifying the native fold and provides physical insight, conceptually linking the results to modern theoretical views of protein folding. In addition to presenting a method for prediction of structure and folding mechanism, our model suggests that an accurate all-atom amino acid representation coupled with a physically reasonable atomic interaction potential and hydrogen bonding are essential features for a realistic protein model. PMID- 16365307 TI - Increasing Ca2+ transients by broadening postsynaptic action potentials enhances timing-dependent synaptic depression. AB - Repeated induction of pre- and postsynaptic action potentials (APs) at a fixed time difference leads to long-term potentiation (LTP) or long-term depression (LTD) of the synapse, depending on the temporal order of pre- and postsynaptic activity. This phenomenon of spike-timing-dependent plasticity (STDP) is believed to arise by nonlinear processes that lead to larger calcium transients (and thus LTP) when presynaptic APs precede postsynaptic APs and smaller calcium transients (and thus LTD) when postsynaptic APs precede presynaptic APs. In contrast to predictions from such calcium-peak-detector models, we show that constitutively or artificially broadened APs in layer II/III pyramidal cells of entorhinal cortex (EC) lead to an increase in the dendritic calcium transient and shift the balance of STDP toward LTD. STDP in entorhinal pyramidal cells is NMDA-receptor dependent and modulated by the Ca(V)1Ca(2+) channel-blocker nifedipine. Results are consistent with an elaboration of the calcium-peak-detector model in which downstream signals from voltage-dependent Ca(2+) channels suppress LTP relative to LTD. Our results suggest that modulation of AP width is a potent way to adjust the rules of synaptic plasticity in the EC. PMID- 16365308 TI - Trans-activation of EphA4 and FGF receptors mediated by direct interactions between their cytoplasmic domains. AB - A yeast two-hybrid analysis has shown that the juxtamembrane region of FGF receptor 3 (FGFR3) interacts with the cytoplasmic domain of EphA4, which is a member of the largest family of receptor tyrosine kinases. Complex formation between the two receptors was shown to be mediated by direct interactions between the juxtamembrane domain of FGFR1, FGFR2, FGFR3, or FGFR4 and the N-terminal portion of the tyrosine kinase domain of EphA4. Activation of FGFR1 in transfected cells resulted in tyrosine phosphorylation of a kinase-negative EphA4 mutant and activation of EphA4 led to tyrosine phosphorylation of a kinase negative FGFR1 mutant. Moreover, both receptors stimulate tyrosine phosphorylation of the docking protein FRS2alpha and induce mitogen-activated protein kinase stimulation with a time course and intensity that depends on the ligand that is applied. We also demonstrate that FGF-receptor-mediated mitogen activated protein kinase stimulation is potentiated in cells costimulated with ephrin-A1. The direct interaction between EphA4 and FGFRs and the potentiation of FGF response that is induced by ephrin-A1 stimulation may modulate the biological responses that are mediated by these receptor families in cells or tissues in which the two receptors are coexpressed. PMID- 16365310 TI - Placing confidence limits on the molecular age of the human-chimpanzee divergence. AB - Molecular clocks have been used to date the divergence of humans and chimpanzees for nearly four decades. Nonetheless, this date and its confidence interval remain to be firmly established. In an effort to generate a genomic view of the human-chimpanzee divergence, we have analyzed 167 nuclear protein-coding genes and built a reliable confidence interval around the calculated time by applying a multifactor bootstrap-resampling approach. Bayesian and maximum likelihood analyses of neutral DNA substitutions show that the human-chimpanzee divergence is close to 20% of the ape-Old World monkey (OWM) divergence. Therefore, the generally accepted range of 23.8-35 millions of years ago for the ape-OWM divergence yields a range of 4.98-7.02 millions of years ago for human-chimpanzee divergence. Thus, the older time estimates for the human-chimpanzee divergence, from molecular and paleontological studies, are unlikely to be correct. For a given the ape-OWM divergence time, the 95% confidence interval of the human chimpanzee divergence ranges from -12% to 19% of the estimated time. Computer simulations suggest that the 95% confidence intervals obtained by using a multifactor bootstrap-resampling approach contain the true value with >95% probability, whether deviations from the molecular clock are random or correlated among lineages. Analyses revealed that the use of amino acid sequence differences is not optimal for dating human-chimpanzee divergence and that the inclusion of additional genes is unlikely to narrow the confidence interval significantly. We conclude that tests of hypotheses about the timing of human-chimpanzee divergence demand more precise fossil-based calibrations. PMID- 16365309 TI - The cannabinoid agonist WIN55,212-2 increases intracellular calcium via CB1 receptor coupling to Gq/11 G proteins. AB - Central nervous system responses to cannabis are primarily mediated by CB(1) receptors, which couple preferentially to G(i/o) G proteins. Here, we used calcium photometry to monitor the effect of CB(1) activation on intracellular calcium concentration. Perfusion with 5 microM CB(1) aminoalkylindole agonist, WIN55,212-2 (WIN), increased intracellular calcium by several hundred nanomolar in human embryonic kidney 293 cells stably expressing CB(1) and in cultured hippocampal neurons. The increase was blocked by coincubation with the CB(1) antagonist, SR141716A, and was absent in nontransfected human embryonic kidney 293 cells. The calcium rise was WIN-specific, being essentially absent in cells treated with other classes of cannabinoid agonists, including Delta(9) tetrahydrocannabinol, HU-210, CP55,940, 2-arachidonoylglycerol, methanandamide, and cannabidiol. The increase in calcium elicited by WIN was independent of G(i/o), because it was present in pertussis toxin-treated cells. Indeed, pertussis toxin pretreatment enhanced the potency and efficacy of WIN to increase intracellular calcium. The calcium increases appeared to be mediated by G(q) G proteins and phospholipase C, because they were markedly attenuated in cells expressing dominant-negative G(q) or treated with the phospholipase C inhibitors U73122 and ET-18-OCH(3) and were accompanied by an increase in inositol phosphates. The calcium increase was blocked by the sarco/endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin, the inositol trisphosphate receptor inhibitor xestospongin D, and the ryanodine receptor inhibitors dantrolene and 1,1' diheptyl-4,4'-bipyridinium dibromide, but not by removal of extracellular calcium, showing that WIN releases calcium from intracellular stores. In summary, these results suggest that WIN stabilizes CB(1) receptors in a conformation that enables G(q) signaling, thus shifting the G protein specificity of the receptor. PMID- 16365311 TI - Evidence by molecular profiling for a placental origin of infantile hemangioma. AB - The origin of the pathogenic endothelial cells in common infantile hemangioma is unknown. We show here that the transcriptomes of human placenta and infantile hemangioma are sufficiently similar to suggest a placental origin for this tumor, expanding on recent immunophenotypical studies that have suggested this possibility [North, P. E., et al. (2001) Arch. Dermatol. 137, 559-570]. The transcriptomes of placenta, hemangioma, and eight normal and diseased tissues were compared by hierarchical and nonhierarchical clustering analysis of >7,800 genes. We found that the level of transcriptome similarity between placenta and hemangioma exceeded that of any other tissue compared and paralleled that observed between a given tissue and its derived tumor, such as normal and cancerous lung. The degree of similarity was even greater when a subset of endothelial cell-specific genes was analyzed. Genes preferentially expressed in both placenta and hemangiomas were identified, including 17-beta hydroxysteroid dehydrogenase type 2 and tissue factor pathway inhibitor 2. These data demonstrate the value of global molecular profiling of tissues as a tool for hypothesis-driven research. Furthermore, it suggests that the unique self-limited growth of infantile hemangioma may, in fact, mirror the lifetime of placental endothelium. PMID- 16365312 TI - On the mechanism of sensing unfolded protein in the endoplasmic reticulum. AB - Unfolded proteins in the endoplasmic reticulum (ER) activate the ER transmembrane sensor Ire1 to trigger the unfolded protein response (UPR), a homeostatic signaling pathway that adjusts ER protein folding capacity according to need. Ire1 is a bifunctional enzyme, containing cytoplasmic kinase and RNase domains whose roles in signal transduction downstream of Ire1 are understood in some detail. By contrast, the question of how its ER-luminal domain (LD) senses unfolded proteins has remained an enigma. The 3.0-A crystal structure and consequent structure-guided functional analyses of the conserved core region of the LD (cLD) leads us to a proposal for the mechanism of response. cLD exhibits a unique protein fold and is sufficient to control Ire1 activation by unfolded proteins. Dimerization of cLD monomers across a large interface creates a shared central groove formed by alpha-helices that are situated on a beta-sheet floor. This groove is reminiscent of the peptide binding domains of major histocompatibility complexes (MHCs) in its gross architecture. Conserved amino acid side chains in Ire1 that face into the groove are shown to be important for UPR activation in that their mutation reduces the response. Mutational analyses suggest that further interaction between cLD dimers is required to form higher order oligomers necessary for UPR activation. We propose that cLD directly binds unfolded proteins, which changes the quaternary association of the monomers in the membrane plane. The changes in the ER lumen in turn position Ire1 kinase domains in the cytoplasm optimally for autophosphorylation to initiate the UPR. PMID- 16365313 TI - Structure of the mid-region of tropomyosin: bending and binding sites for actin. AB - Tropomyosin is a two-chain alpha-helical coiled coil whose periodic interactions with the F-actin helix are critical for thin filament stabilization and the regulation of muscle contraction. Here we deduce the mechanical and chemical basis of these interactions from the 2.3-A-resolution crystal structure of the middle three of tropomyosin's seven periods. Geometrically specific bends of the coiled coil, produced by clusters of core alanines, and variable bends about gaps in the core, produced by isolated alanines, occur along the molecule. The crystal packing is notable in signifying that the functionally important fifth period includes an especially favorable protein-binding site, comprising an unusual apolar patch on the surface together with surrounding charged residues. Based on these and other results, we have constructed a specific model of the thin filament, with the N-terminal halves of each period (i.e., the so-called "alpha zones") of tropomyosin axially aligned with subdomain 3 of each monomer in F actin. PMID- 16365314 TI - Correlation between evolutionary structural development and protein folding. AB - Evolution should have played important roles in determining folding mechanisms and structures of proteins. In this article we discuss how the folding mechanisms had been affected by the early stage of evolution through which the uniqueness of structure had developed. Although the process of such early-time evolution has remained a mystery, a plausible scenario is that the evolution of proteins toward the ordered structures was guided by functional selection pressure as demonstrated in vitro and in silico. We examine the in silico functional selection of sequences and show that there is a significant correlation between two different processes toward the unique 3D structure, the evolutionary development of structure through sequence selection, and the folding process of the resultant sequence. This finding could be rephrased as protein folding recapitulates the emergence of topology in the molecular evolution. The correlation suggests a guideline for engineering foldable proteins. PMID- 16365315 TI - Class II-restricted T cell receptor engineered in vitro for higher affinity retains peptide specificity and function. AB - The T cell receptor (TCR) alphabeta heterodimer determines the peptide and MHC specificity of a T cell. It has been proposed that in vivo selection processes maintain low TCR affinities because T cells with higher-affinity TCRs would (i) have reduced functional capacity or (ii) cross-react with self-peptides resulting in clonal deletion. We used the class II-restricted T cell clone 3.L2, specific for murine hemoglobin (Hb/I-E(k)), to explore these possibilities by engineering higher-affinity TCR mutants. A 3.L2 single-chain TCR (Vbeta-linker-Valpha) was mutagenized and selected for thermal stability and surface expression in a yeast display system. Stabilized mutants were used to generate a library with CDR3 mutations that were selected with Hb/I-E(k) to isolate a panel of affinity mutants with K(D) values as low as 25 nM. Kinetic analysis of soluble single chain TCRs showed that increased affinities were the result of both faster on rates and slower off-rates. T cells transfected with the mutant TCRs and wild type TCR responded to similar concentrations of peptide, indicating that the increased affinity was not detrimental to T cell activation. T cell transfectants maintained exquisite hemoglobin peptide specificity, but an altered peptide ligand that acted as an antagonist for the wild-type TCR was converted to a strong agonist with higher-affinity TCRs. These results show that T cells with high-affinity class II reactive TCRs are functional, but there is an affinity threshold above which an increase in affinity does not result in significant enhancement of T cell activation. PMID- 16365316 TI - Abnormal centrosome amplification in cells through the targeting of Ran-binding protein-1 by the human T cell leukemia virus type-1 Tax oncoprotein. AB - Human T cell leukemia virus type-1 (HTLV-1) is an oncogenic retrovirus etiologically causal of adult T cell leukemia. The virus encodes a Tax oncoprotein that functions in transcriptional regulation, cell cycle control, and transformation. Because adult T cell leukemia like many other human cancers is a disease of genomic instability with frequent gains and losses of chromosomes, to understand this disease it is important to comprehend how HTLV-1 engenders aneuploidy in host cells. In this regard, loss of cell cycle checkpoints permits tolerance of aneuploidy but does not explain how aneuploidy is created. We show here that HTLV-1 Tax causes abnormal centrosome fragmentation in the mitotic phase of the cell cycle. We report that Tax directly binds Ran and Ran-binding protein-1, locates to centrosomes/spindle poles, and causes supernumerary centrosomes. PMID- 16365317 TI - Opioid-induced tolerance and dependence in mice is modulated by the distance between pharmacophores in a bivalent ligand series. AB - Given the mounting evidence for involvement of delta opioid receptors in the tolerance and physical dependence of mu opioid receptor agonists, we have investigated the possible physical interaction between mu and delta opioid receptors by using bivalent ligands. Based on reports of suppression of antinociceptive tolerance by the delta antagonist naltrindole (NTI), bivalent ligands [mu-delta agonist-antagonist (MDAN) series] that contain different length spacers, and pharmacophores derived from NTI and the mu agonist oxymorphone, have been synthesized and evaluated by intracerebroventricular (i.c.v.) administration in the tail-flick test in mice. In acute i.c.v. studies, the bivalent ligands functioned as agonists with potencies ranging from 1.6- to 45-fold greater than morphine. In contrast, the monovalent mu agonist analogues were substantially more potent than the MDAN congeners and were essentially equipotent with one another and oxymorphone. Pretreatment with NTI decreased the ED(50) values for MDAN-19 to a greater degree than for MDAN-16 but had no effect on MDAN-21. Chronic i.c.v. studies revealed that MDAN ligands whose spacer was 16 atoms or longer produced less dependence than either morphine or mu monovalent control MA 19. On the other hand, both physical dependence and tolerance were suppressed at MDAN spacer lengths of 19 atoms or greater. These data suggest that physical interaction between the mu and delta opioid receptors modulates mu-mediated tolerance and dependence. Because MDAN-21 was found to be 50-fold more potent than morphine by the i.v. route (i.v.), it offers a previously uncharacterized approach for the development of analgesics devoid of tolerance and dependence. PMID- 16365318 TI - The glycosphingolipid globotriaosylceramide in the metastatic transformation of colon cancer. AB - The most devastating aspect of cancer is the emergence of metastases. Thus, identification of potentially metastatic cells among a tumor cell population and the underlying molecular changes that switch cells to a metastatic state are among the most important issues in cancer biology. Here we show that, although normal human colonic epithelial cells lack the glycosphingolipid globotriaosylceramide (Gb(3)), this molecule is highly expressed in metastatic colon cancer. In addition, a subpopulation of cells that are greatly enriched in Gb(3) and have an invasive phenotype was identified in human colon cancer cell lines. In epithelial cells in culture, Gb(3) was necessary and sufficient for cell invasiveness. Transfection of Gb(3) synthase, resulting in Gb(3) expression in noncancerous polarized epithelial cells lacking endogenous Gb(3), induced cell invasiveness. Furthermore, Gb(3) knockdown by small inhibitory RNA in colon cancer epithelial cells inhibited cell invasiveness. Gb(3) is the plasma membrane receptor for Shiga toxin 1. The noncatalytic B subunit of Shiga toxin 1 causes apoptosis of human colon cancer cells expressing Gb(3). Injections of the B subunit of Shiga toxin 1 into HT29 human colon cancer cells engrafted into the flanks of nude mice inhibited tumor growth. These data demonstrate the appearance of a subpopulation of Gb(3) containing epithelial cells in the metastatic stage of human colon cancer and suggest their possible role in colon cancer invasiveness. PMID- 16365319 TI - Evidence for an essential function of the N terminus of a small heat shock protein in vivo, independent of in vitro chaperone activity. AB - To investigate the mechanism of small heat shock protein (sHsp) function, unbiased by current models of sHsp chaperone activity, we performed a screen for mutations of Synechocystis Hsp16.6 that reduced the ability of the protein to provide thermotolerance in vivo. Missense mutations at 17 positions throughout the protein and a C-terminal truncation of 5 aa were identified, representing the largest collection of sHsp mutants impaired in function in vivo. Ten mutant proteins were purified and tested for alterations in native oligomeric structure and in vitro chaperone activity. These biochemical assays separated the mutants into two groups. The C-terminal truncation and six mutations in the alpha crystallin domain destabilized the sHsp oligomer and reduced in vitro chaperone activity. In contrast, the other three mutations had little effect on oligomer stability or chaperone activity in vitro. These mutations were clustered in the N terminus of Hsp16.6, pointing to a previously unrecognized, important function for this evolutionarily variable domain. Furthermore, the fact that the N terminal mutations were impaired in function in vivo, but active as chaperones in vitro, indicates that current biochemical assays do not adequately measure essential features of the sHsp mechanism of action. PMID- 16365321 TI - Application of the TIMI risk score for unstable angina and non-ST elevation acute coronary syndrome to an unselected emergency department chest pain population. AB - OBJECTIVES: Patients presenting with chest pain or related symptoms suggestive of myocardial ischemia, without ST-segment elevation (NSTE) on their presenting electrocardiograms, often present a diagnostic challenge in the emergency department (ED). Prompt and accurate risk stratification to identify those patients with NSTE chest pain who are at highest risk for adverse events is essential, however, to optimal management. Although validated and used frequently in patients already enrolled in acute coronary syndrome trials, the Thrombolysis in Myocardial Infarction (TIMI) risk score never has been examined for its value in risk stratification in an all-comers, non-trial-based ED chest pain population. METHODS: An analysis of an ED-based prospective observational cohort study was conducted in 3,929 adult patients presenting with chest pain syndrome and warranting evaluation with an electrocardiogram. These patients had TIMI risk scores determined at ED presentation. The main outcome was the composite of death, acute myocardial infarction (MI), and revascularization within 30 days. RESULTS: The TIMI risk score at ED presentation successfully risk-stratified this unselected cohort of chest pain patients with respect to 30-day adverse outcome, with a range from 2.1%, with a score of 0, to 100%, with a score of 7. The highest correlation of an individual TIMI risk indicator to adverse outcome was for elevated cardiac biomarker at admission. Overall, the score had similar performance characteristics to that seen when applied to other databases of patients enrolled in clinical trials and registries using a 14-day end point. CONCLUSIONS: The TIMI risk score may be a useful tool for risk stratification of ED patients with chest pain syndrome. PMID- 16365320 TI - P2Y2 receptor activates nerve growth factor/TrkA signaling to enhance neuronal differentiation. AB - Neurotrophins are essential for neuronal differentiation, but the onset and the intensity of neurotrophin signaling within the neuronal microenvironment are poorly understood. We tested the hypothesis that extracellular nucleotides and their cognate receptors regulate neurotrophin-mediated differentiation. We found that 5'-O-(3-thio)triphosphate (ATPgammaS) activation of the G protein-coupled receptor P2Y(2) in the presence of nerve growth factor leads to the colocalization and association of tyrosine receptor kinase A and P2Y(2) receptors and is required for enhanced neuronal differentiation. Consistent with these effects, ATPgammaS promotes phosphorylation of tyrosine receptor kinase A, early response kinase 1/2, and p38, thereby enhancing sensitivity to nerve growth factor and accelerating neurite formation in both PC12 cells and dorsal root ganglion neurons. Genetic or small interfering RNA depletion of P2Y(2) receptors abolished the ATPgammaS-mediated increase in neuronal differentiation. Moreover, in vivo injection of ATPgammaS into the sciatic nerve increased growth-associated protein-43 (GAP-43), a marker for axonal growth, in wild-type but not P2Y(2)(-/-) mice. The interactions of tyrosine kinase- and P2Y(2)-signaling pathways provide a paradigm for the regulation of neuronal differentiation and suggest a role for P2Y(2) as a morphogen receptor that potentiates neurotrophin signaling in neuronal development and regeneration. PMID- 16365322 TI - Out-of-hospital auricular acupressure in elder patients with hip fracture: a randomized double-blinded trial. AB - OBJECTIVES: Auricular acupressure is known to decrease the level of anxiety in patients during ambulance transport. The purpose of this randomized, double blind, sham control study was to determine whether auricular acupressure can decrease not only the level of anxiety but also the level of pain in a group of elder patients with acute hip fracture. METHODS: With the assistance of the Vienna Red Cross, 38 patients with acute hip fracture were enrolled into this study. Patients were randomized into two study groups: the true intervention group and the sham control group. Subjects in the true intervention group (n = 18) received bilateral auricular acupressure at three auricular acupressure points for hip pain. Patients in the sham group (n = 20) received bilateral auricular acupressure at sham points. Baseline demographic information, anxiety level, pain level, blood pressure, and heart rate were obtained before the administration of the appropriate acupressure intervention. The level of anxiety, level of pain, hemodynamic profiles, and level of satisfaction were reassessed once the patients arrived at the hospital. RESULTS: Patients in the true intervention groups had less pain (F = 28, p = 0.0001) and anxiety (F = 4.3, p = 0.018) and lower heart rate (F = 18, p = 0.0001) on arrival at the hospital than did patients in the sham control group. As a result, the patients in the true intervention group reported higher satisfaction in the care they received during the ride to the hospital. CONCLUSIONS: The authors encourage physicians, health care providers, and emergency rescuers to learn this easy, noninvasive, and inexpensive technique for its effects in decreasing anxiety and pain during emergency transportation. PMID- 16365323 TI - Lazaroid U-83836E improves tolerance to hemorrhagic shock and limb ischemia and reperfusion in rats and increases cardiac heat shock protein 72. AB - OBJECTIVES: Aminosteroids of the lazaroid type protect organs from ischemia reperfusion damage. The authors hypothesized that lazaroid U-83836E may be beneficial in a shock model with hemorrhage combined with limb ischemia. Furthermore, the authors hypothesized that lazaroids induce expression of heat shock proteins (HSPs) of the 72-kDa family. METHODS: Rats were divided into two groups (lazaroid and control groups, n = 8 each) and pretreated with the lazaroid U-83836E (5 mg/kg) or with vehicle intraperitoneally at 12 and 24 hours before experiments. At the time of the experiment, rats were anesthetized, and the femoral artery of each rat was cannulated. After 20 minutes of stabilization, blood was shed from each rat to bring its mean arterial pressure to 24-28 mmHg for 2 hours. Bilateral tourniquets were tightened proximally on the rat thighs during those 2 hours and then released. Shed blood plus equal amounts of Ringer acetate then were infused to restore normal blood pressure, followed by a continuous infusion of Ringer acetate, the rate of which was regulated to maintain blood pressure, until 30 minutes after start of resuscitation. Fluid resuscitation was stopped, and rats were observed for another 3.5 hours. At the end of the observation period, the rats' hearts were collected for immunoblot analysis of HSP72. Additional hearts were collected from similarly pretreated rats not undergoing the episode of hemorrhagic shock and fluid resuscitation. RESULTS: Pretreatment with U-83836E improved mean arterial blood pressure after hemorrhagic shock and fluid resuscitation (p = 0.02), combined with improvements in acid-base balance (improved base excess and standard bicarbonate; p = 0.02 and p = 0.01, respectively). Western blot of cardiac protein extracts demonstrated that lazaroid pretreatment increased expression of HSP72. CONCLUSIONS: Pretreatment with the lazaroid U-83836E improved outcome markers in this hemorrhagic shock model. The observed protection may be caused by increased expression of HSP72. PMID- 16365324 TI - Description and evaluation of a pilot physician-directed emergency medical services diversion control program. AB - OBJECTIVES: To describe the characteristics and feasibility of a physician directed ambulance destination-control program to reduce emergency department (ED) overcrowding, as measured by hospital ambulance diversion hours. METHODS: This controlled trial took place in Rochester, New York and included a university hospital and a university-affiliated community hospital. During July 2003, emergency medical services (EMS) providers were asked to call an EMS destination control physician for patients requesting transport to either hospital. The destination-control physician determined the optimal patient destination by using patient and system variables as well as EMS providers' and patients' input. Program process measures were evaluated to characterize the program. Administrative data were reviewed to compare system characteristics between the intervention program month and a control month. RESULTS: During the intervention month, 2,708 patients were transported to the participating hospitals. EMS providers contacted the destination-control physician for 1,866 (69%) patients. The original destination was changed for 253 (14%) patients. Reasons for redirecting patients included system needs, patient needs, physician affiliation, recent ED or hospital care, patient wishes, and primary care physician wishes. During the intervention month, EMS diversion decreased 190 (41%) hours at the university hospital and 62 (61%) hours at the community hospital, as compared with the control month. CONCLUSIONS: A voluntary, physician-directed destination control program that directs EMS units to the ED most able to provide appropriate and timely care is feasible. Patients were redirected to maximize continuity of care and optimally use available emergency health care resources. This type of program may be effective in reducing overcrowding. PMID- 16365325 TI - Profiles in patient safety: confirmation bias in emergency medicine. AB - Confirmation bias is a pitfall in emergency care and may lead to inaccurate diagnoses and inappropriate treatments and care plans. Because of the increasing severity and volume of emergency care, emergency physicians often must rely on heuristics, such as rule-out protocols, as a guide to diagnosing and treating patients. The use of heuristics or protocols can be potentially misleading if the initial diagnostic impression is incorrect. To minimize cognitive dissonance, clinicians may accentuate confirmatory data and ignore nonconfirmatory data. Clinicians should recognize confirmation bias as a potential pitfall in medical decision making in the emergency department. Reliance on the scientific method, Bayesian reasoning, metacognition, and cognitive forcing strategies may serve to improve diagnostic accuracy and improve patient care. PMID- 16365326 TI - Validation of probabilistic linkage to match de-identified ambulance records to a state trauma registry. AB - OBJECTIVES: To validate the accuracy of using probabilistic linkage for matching de-identified ambulance records to a state trauma registry. METHODS: This was a retrospective cohort analysis. Three thousand nine hundred nineteen true matches between ambulance and state trauma registry data from 1998 to 2003 were identified by deterministic matching on trauma identification number and verified by human review. Two thousand thirty-eight ambulance records from trauma patients not meeting criteria for a true match, and an identical number of trauma registry records randomly selected from the one local county served by a different EMS provider, were included as nonmatches. There were 17 variables considered for linkage, which included the following: age, gender, race, county, hospital, date, rural setting, call and arrival times, mechanism, penetrating injury, vital signs, intubation, and intoxication. Probabilistic linkage was used to link the two data sets, using seven different combinations of common variables (maximum, 17; minimum, 4). The sensitivity and specificity of identifying true matches and nonmatches (95% confidence intervals [95% CI]) were calculated for each combination of variables. RESULTS: Using all 17 available variables, 3,766 of 3,919 true matches were appropriately linked (sensitivity, 96.1%; 95% CI = 95.4% to 96.7%), with eight mismatches (specificity, 99.6%; 95% CI = 99.2% to 99.8%). Sensitivity fell below 95% with < 15 variables; however, sensitivity was very dependent on the inclusion of variables with high discriminatory power. Specificity remained >98% regardless of the number of variables included. CONCLUSIONS: Probabilistic linkage is a valid method for matching ambulance records to a trauma registry without the use of patient identifiers; however, the sensitivity of identifying true matches is critically dependent on the number and type of common variables included in the analysis. PMID- 16365327 TI - Predicting smoking stage of change among emergency department patients and visitors. AB - OBJECTIVES: Little is known about whether emergency department (ED) patients or those who accompany them (visitors) are interested in smoking cessation. The authors hypothesized that several variables would be associated with stage of change, including nicotine dependence, self-efficacy, presence of a smoking related illness, and anticipated cessation-related health improvement. METHODS: For two 24-hour periods, consecutive patients and visitors aged 18 years and older presenting to four Boston EDs were interviewed. The authors assessed a range of smoking-related constructs. Exclusion criteria included severe illness, cognitive insufficiency, and acute distress. RESULTS: One thousand ten subjects were screened (56% patients, 44% visitors). Two hundred thirty-seven (23%) subjects were current smokers, with 57% being in precontemplation, 31% in contemplation, and 12% in preparation stages. When ordinal regression was used, the variables most strongly associated with stage of change were as follows: self efficacy (odds ratio [OR] = 5.1; p < 0.001), anticipated cessation-related health improvement (OR = 2.7; p = 0.02), and having a smoking-related health problem (OR = 1.9; p = 0.08). CONCLUSIONS: Because many disenfranchised Americans use the ED as a regular source of health care, increased attention to smoking in the ED setting holds tremendous public health potential. This study's results reinforce the validity of the stage-of-change model within the ED setting. Developers of ED initiated interventions will have to consider the heterogeneity in stage of change when designing their treatments. PMID- 16365328 TI - Susceptibility of recent bacterial isolates to cefdinir and selected antibiotics among children with urinary tract infections. AB - BACKGROUND: Cefdinir, an extended-spectrum cephalosporin administered orally, is approved by the U.S. Federal Drug Administration for treatment of skin and respiratory tract infections. During the last two years at the authors' institution, this agent has been used as an off-label treatment for urinary tract infections in children. OBJECTIVES: To evaluate antimicrobial susceptibility testing data in children to determine whether there is support for this prescribing practice. METHODS: In this retrospective study (2003-2004), the authors compared the susceptibility patterns of urinary pathogens to cefdinir and selected antibiotics in children who were evaluated for urinary tract infections in an urban tertiary academic pediatric emergency department. Pathogens (community acquired vs. opportunistic or nosocomial) were categorized as susceptible, indeterminate, or resistant on the basis of antibiotic susceptibility breakpoints. The frequency of these categorizations for individual drugs was determined. RESULTS: Seven hundred five isolates were recovered from urine during the study period. Pathogens isolated most frequently were Escherichia coli, Klebsiella spp, and Proteus spp. Of 431 isolates retained in the data set, 412 (95.6%) were susceptible to cefdinir. This rate was comparable or superior to rates observed for other antibiotics: 49.4% for ampicillin, 84.9% for trimethoprim-sulfamethoxazole, 88.4% for cefazolin, 93.3% for nitrofurantoin, 94.2% for ticarcillin-clavulanate potassium, 97.5% for gentamicin, and 97.7% for ceftriaxone. Cefdinir, however, had lower activity (64.7%) against 17 bacterial isolates categorized as opportunistic or nosocomial pathogens. CONCLUSIONS: Cefdinir provides good coverage against common pathogens responsible for urinary tract infections in children and compares favorably with other oral and parenteral antibiotics that are used in the empiric treatment of this infection. PMID- 16365329 TI - Using queueing theory to increase the effectiveness of emergency department provider staffing. AB - OBJECTIVES: Significant variation in emergency department (ED) patient arrival rates necessitates the adjustment of staffing patterns to optimize the timely care of patients. This study evaluated the effectiveness of a queueing model in identifying provider staffing patterns to reduce the fraction of patients who leave without being seen. METHODS: The authors collected detailed ED arrival data from an urban hospital and used a Lag SIPP queueing analysis to gain insights on how to change provider staffing to decrease the proportion of patients who leave without being seen. The authors then compared this proportion for the same 39 week period before and after the resulting changes. RESULTS: Despite an increase in arrival volume of 1,078 patients (6.3%), an average increase in provider hours of 12 hours per week (3.1%) resulted in 258 fewer patients who left without being seen. This represents a decrease in the proportion of patients who left without being seen by 22.9%. Restricting attention to a four-day subset of the week during which there was no increase in total provider hours, a reallocation of providers based on the queueing model resulted in 161 fewer patients who left without being seen (21.7%), despite an additional 548 patients (5.5%) arriving in the second half of the study. CONCLUSIONS: Timely access to a provider is a critical dimension of ED quality performance. In an environment in which EDs are often understaffed, analyses of arrival patterns and the use of queueing models can be extremely useful in identifying the most effective allocation of staff. PMID- 16365330 TI - Influence of electromagnetic fields on function of automated external defibrillators. AB - OBJECTIVES: In this study, the authors tested whether electromagnetic interference (EMI) is able to impair correct electrocardiogram analysis and produce false-positive shock advice from automated external defibrillators (AEDs) when the true rhythm is sinus. METHODS: Nineteen healthy subjects were used to test five AEDs available on the Austrian market in a prospective, open, and sequence-randomized study. The primary outcome variable was the absolute number of shocks advised in the presence of EMI. The secondary outcome was the number of impaired analyses caused by incorrectly detected patient movements or electrode failure. RESULTS: Of 760 tests run, 18 (2.37%) cases of false-positive results occurred, and two of five AEDs recommended shocks in the presence of sinus rhythm. Of 760 tests run, no electrode failures occurred. There were 27 occurrences (3.55%) of motion detected by an AED in the presence of strong electromagnetic fields. CONCLUSIONS: AED models differ in their response to EMI; it may be useful to consider specific safety requirements for areas with such fields present. Working personnel and emergency medical services staff should be informed about potential risks and the possible need for patient evacuation before AEDs are attached and shock recommendations are followed. PMID- 16365331 TI - Accuracy of mild traumatic brain injury case ascertainment using ICD-9 codes. AB - OBJECTIVES: To determine the accuracy of mild traumatic brain injury (TBI) case ascertainment using International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) codes proposed by the Centers for Disease Control and Prevention (CDC) in a 2003 Report to Congress. METHODS: This was a prospective cohort study of all patients presenting to an urban academic emergency department (ED) over six months in 2003. A real-time clinical assessment of mild TBI was compared with the ICD-9 codes assigned after ED or hospital discharge for a determination of sensitivity and specificity. RESULTS: Of the 35,096 patients presenting to the ED, 516 had clinically defined mild TBI and 1,000 were assigned one or more of the mild TBI ICD-9 codes proposed by the CDC. The sensitivity of these codes was 45.9% (95% confidence interval [95% CI] = 41.3% to 50.2%) with a specificity of 97.8% (95% CI = 97.6% to 97.9%). CONCLUSIONS: The identification of mild TBI patients using retrospectively assigned ICD-9 codes appears to be inaccurate. These codes are associated with a significant number of false-positive and false-negative code assignments. Mild TBI incidence and prevalence estimates using these codes should be interpreted with caution. ICD-9 codes should not replace a clinical assessment for mild TBI when accurate case ascertainment is required. PMID- 16365332 TI - Publication bias of randomized controlled trials in emergency medicine. AB - OBJECTIVES: To determine the publication status and time to publication of randomized controlled trials (RCTs) that were presented at the Society for Academic Emergency Medicine (SAEM) meetings from 1995 to 2003. The impact of positive-outcome bias, time-lag bias, and gray literature bias also was assessed. METHODS: Retrospective cohort study of RCT abstracts presented at nine SAEM scientific meetings. Electronic searches identified publications from the abstracts. RESULTS: Of 4,399 abstracts, 383 (8.7%; 95% confidence interval [CI] = 7.8% to 9.5%) were identified as RCTs. One hundred ninety-four (50.7%; 95% CI = 45.7% to 55.7%) were subsequently published up to May 2004. The median time to publication was 32 months (95% CI = 23 to 41), with 59% of RCT abstracts published within five years of presentation. No evidence of positive-outcome bias or time-lag bias was identified; however, changes from abstract to manuscript were found. Manuscripts were less likely to endorse the experimental intervention than were abstracts (OR, 0.2; 95% CI = 0.0 to 0.6). CONCLUSIONS: The proportion of emergency medicine RCT abstracts published is slightly lower than that for other biomedical specialties; however, biases reported by investigators in other biomedical areas do not appear to be as problematic in emergency medicine research. Differences between conclusions from abstracts and manuscripts must be considered when employing meeting abstracts as a source of evidence for future research or for systematic reviews in emergency medicine. PMID- 16365333 TI - A test to detect chest tube kinking. AB - OBJECTIVES: To determine whether the Mac-technique test can detect kinking of the chest tube upon thoracostomy tube placement. METHODS: This was a prospective observational study that was conducted October 2000 through October 2001 in an urban Level 1 trauma center. There were 103 consecutive nonrandomized adult trauma patients who required immediate tube thoracostomy during their initial resuscitation who were entered into the study. The Mac-technique test was performed during standard tube thoracostomy insertion to the appropriate depth. The test involved grasping the external portion of the thoracostomy tube, turning it clockwise 180 degrees, and then releasing the tube. If the tube spontaneously spun back to its original position, the test was considered positive, and the tube was considered kinked. If the tube did not spontaneously spin back and stayed in position upon release, the test was considered negative. Regardless of the results of this test, the tube was secured, and a postprocedure chest radiograph was obtained. The criterion standard for determining a kinked chest tube was its appearance on this chest radiograph. RESULTS: A total of 103 chest tubes were placed by using the Mac-technique test. The test was positive in eight placements; four tubes were kinked on chest radiograph. The Mac-technique test was negative in 95 placements; four tubes were kinked on chest radiograph. The Mac technique had a sensitivity of 50% (95% confidence interval [CI] = 15.7% to 84.3%), a specificity of 95.8% (95% CI = 89.6% to 98.8%), a positive likelihood ratio of 11.9, a negative likelihood ratio of 0.52, and an odds ratio using Yates correction of 20.3 (95% CI = 4.1 to 102.1). CONCLUSIONS: On the basis of this study, a positive Mac-technique test is useful to detect chest tubes that are likely to be kinked after insertion and before securing. PMID- 16365334 TI - Prehospital 12-lead electrocardiography impact on acute myocardial infarction treatment times and mortality: a systematic review. AB - OBJECTIVES: Prehospital 12-lead electrocardiogram (PHECG) interpretation and advance emergency department (ED) notification may improve time-to-treatment intervals for a variety of treatment strategies to improve outcome in acute myocardial infarction. Despite consensus guidelines recommending this intervention, few emergency medical services (EMS) employ this. The authors systematically reviewed the literature to report whether mortality or treatment time intervals improved when compared with standard care. METHODS: The authors used the Cochrane strategy to search MEDLINE, EMBASE, Current Contents, Dissertation Abstracts, Cochrane Library, and Index of Scientific and Technical Proceedings. Bibliographies and grant-agency Websites were reviewed, and primary investigators and industry were contacted for published and unpublished studies. Inclusion criteria included PHECG and advance ED notification versus standard EMS care; controlled trials; English only; and evaluation of treatment time intervals, all-cause mortality, or both. Study selection was hierarchical, blinded, and independent. Agreement at each level of review was evaluated by using a kappa statistic. Study quality was measured with a validated scale and was interpreted by two independent reviewers. RESULTS: A total of 1,283 citations were identified, and five studies met the inclusion criteria. The weighted kappa for selection was 0.61 (standard error [SE], 0.045) for titles, 0.63 (SE, 0.051) for abstracts, and 0.79 (SE, 0.146) for full articles. Mean study quality measures by two independent reviewers were 6.0/15 and 5.5/15 (correlation coefficient, 0.85; p = 0.06). PHECG and advance ED notification increased the weighted mean on-scene time by 1.2 minutes (95% confidence interval [95% CI] = 0.84 to 3.2). The weighted mean door-to-needle interval was shortened by 36.1 minutes (95% CI = 9.3 to 63.0: range of means, 22-48 minutes vs. 50-97 minutes). One study reported all-cause mortality, with a statistically nonsignificant reduction from 15.6% to 8.4%. CONCLUSIONS: For patients with AMI, the literature would suggest that PHECG and advanced ED notification reduces in hospital time to fibrinolysis. One controlled trial found no difference in mortality with this out of-hospital intervention. PMID- 16365335 TI - Funding and publishing trends of original research by emergency medicine investigators over the past decade. AB - OBJECTIVES: To update the profile of author-reported funding of reports of original research published since 1994 in the four U.S. peer-reviewed general emergency medicine (EM) journals. METHODS: For the primary analysis, articles published in Academic Emergency Medicine, American Journal of Emergency Medicine, Annals of Emergency Medicine, and Journal of Emergency Medicine between 1994 and 2003 were reviewed by two emergency physicians trained in abstracting from the literature. Original research was identified; case reports, case series, abstracts, and meta-analysis were excluded. Articles were classified by author report of extramural funding as government (public), private nonprofit (foundation), for profit (industry), or multiple. Data are reported as percent per year per journal. A secondary analysis of MEDLINE was performed to identify publishing trends by EM investigators outside of these four journals over the same period. RESULTS: The primary analysis identified 5,728 articles; 3,278 (57%) were considered original research, with 32% of these reporting extramural funding. In 1994, there were 292 articles with 28% funded (28% public, 43% foundation, 17% industry, and 12% multiple). By 2003, there were 358 articles with 36% funded (42% public, 30% foundation, 18% industry, and 10% multiple). There was a trend toward increased extramural funding in all four journals (p = 0.007), with an estimated growth of 3.6% (95% confidence interval = 1.0% to 6.3%) per year over the past decade. The primary review identified 48% of all articles published by EM investigators. CONCLUSIONS: Author-reported extramural funding rates for original research have increased in the EM literature over the past decade. Foundations have funded the largest number of studies, with public (government) sources increasing in 2003. PMID- 16365336 TI - Early goal-directed therapy, corticosteroid, and recombinant human activated protein C for the treatment of severe sepsis and septic shock in the emergency department. AB - OBJECTIVES: To describe our experience with early goal-directed therapy (EGDT), corticosteroid administration, and recombinant human activated protein C (rhAPC) administration in patients with severe sepsis or septic shock and an Acute Physiology and Chronic Health Evaluation (APACHE) II score > or =25 in the emergency department (ED). METHODS: This was a retrospective case series of a prospectively maintained ED sepsis registry. Data are presented as median (25th, 75th percentile). The setting was an academic tertiary ED with approximately 60,000 annual patient visits. Patients with severe sepsis or septic shock and an APACHE II score > or =25 entered in an ED sepsis registry over a four-month period were included. Patients who received rhAPC in the intensive care unit were excluded. Central venous catheterization for central venous pressure and central venous oxygen saturation monitoring, antibiotics, fluid resuscitation, mechanical ventilation, vasopressors, inotropes, corticosteroids, and rhAPC were initiated by the emergency physicians and continued in the intensive care unit by intensivists. RESULTS: Twenty-four patients were enrolled. Patient characteristics were as follows: age, 79.5 (68.0, 83.5) years; APACHE II score, 31.5 (29.8, 36.0); ED length of stay, 6.5 (4.0, 10.5) hours; predicted mortality, 76.7% (71.9, 86.4); and in-hospital mortality, 45.8%. All patients received broad spectrum antibiotics, 54.2% completed EGDT, 33.3% received corticosteroids, and 33.3% received rhAPC. Time of antibiotic administration was 1.5 (1.0, 2.0) hours, time of central venous pressure/central venous oxygen saturation monitoring was 1.0 (0.5, 2.5) hour, and time of rhAPC administration was 9.5 (6.8, 10.5) hours after patients met criteria for severe sepsis or septic shock. In-hospital mortality of patients who received rhAPC in addition to other therapies was 25.0%. CONCLUSIONS: EGDT, corticosteroid administration, and rhAPC administration are feasible in the ED setting. While these evidence-based therapies individually have been shown to improve outcomes for patients with severe sepsis or septic shock, further studies are needed to examine their combined effectiveness during the early stages of this disease. PMID- 16365337 TI - Propofol for emergency department procedural sedation and analgesia: a tale of three centers. AB - OBJECTIVES: To characterize propofol procedural sedation and analgesia (PSA) encounters for a large patient population at multiple emergency department (ED) sites. The authors sought to assess the frequency of respiratory and cardiovascular events during propofol PSA within these settings. METHODS: This study was a prospective, descriptive series of a consecutive sample of ED patients receiving propofol for PSA at three study sites. Patients were monitored for PSA-related events, including predefined clinically relevant cardiovascular and respiratory events. Data collection was performed during PSA with a standardized data collection sheet unique to each site. RESULTS: Propofol was administered during PSA to 792 patients during the respective reporting period at each center. Indications for sedation included dislocation reduction (38%), cardioversion (10%), fracture reduction (35%), abscess incision and drainage (8%), computed tomography imaging (2%), and tube thoracostomy (1%). The cumulative rate of oxygen desaturation events for all study sites was 7.7% with a brief period of assisted ventilation with bag-valve mask in 3.9%. The cumulative rate of PSA-related hypotensive events was 3.5%. Increasing patient age and specific clinical procedure were clinical variables most associated with any propofol-related respiratory event. All PSA-related events resolved with supportive interventions during the PSA encounter. No patients required endotracheal intubation, prolonged observation, or admission for PSA-related complications. CONCLUSIONS: Propofol typically confers a deep sedation experience for ED PSA. The most common PSA events associated with propofol are respiratory related and appear consistent across these three practice settings. All propofol related PSA events resolved with brief supportive interventions in the ED with no adverse sequelae. PMID- 16365338 TI - Simulation-based morbidity and mortality conference: new technologies augmenting traditional case-based presentations. AB - The authors describe the use of a high-fidelity simulation laboratory to re create a patient encounter for the purposes of enhancing a morbidity and mortality conference. The use of two separate technologies were enlisted: a METI high-fidelity patient simulator to re-create the case in a more lifelike fashion, and an audience response system to collect clinical impressions throughout the case presentation and survey data at the end of the presentation. The re-creation of the patient encounter with all relevant physical findings displayed in high fidelity, with relevant laboratory data, nursing notes, and imaging as it occurred in the actual case, provides a more engaging format for the resident learner. This technological enhancement was deployed at a morbidity and mortality conference, and the authors report the impressions collected via the audience response system. Guidelines for those who wish to re-create this type of educational experience are presented in the discussion. PMID- 16365339 TI - Evidence-based therapies for sepsis care in the emergency department: striking a balance between feasibility and necessity. PMID- 16365340 TI - Percutaneous coronary intervention: recommendations for good practice and training. AB - Cardiologists undertaking percutaneous coronary intervention (PCI) are excited by the combination of patient and physician satisfaction and technological advance occurring on the background of the necessary manual dexterity. Progress and applicability of percutaneous techniques since their inception in 1977 have been remarkable; a sound evidence base coupled with the enthusiasm and ingenuity of the medical device industry has resulted in a sea change in the treatment of coronary heart disease (CHD), which continues to evolve at breakneck speed. This is the third set of guidelines produced by the British Cardiovascular Intervention Society and the British Cardiac Society. Following the last set of guidelines published in 2000, we have seen PCI activity in the UK increase from 33,652 to 62,780 (87% in four years) such that the PCI to coronary artery bypass grafting ratio has increased to 2.5:1. The impact of drug eluting stents has been profound, and the Department of Health is investigating the feasibility of primary PCI for acute myocardial infarction. Nevertheless, the changes in the structure of National Health Service funding are likely to focus our attention on cost effective treatments and will require physician engagement and sensitive handling if we are to continue the rapid and appropriate growth in our chosen field. It is important with this burgeoning development now occurring on a broad front (in both regional centres and district general hospitals) that we maintain our vigilance on audit and outcome measures so that standards are maintained for both operators and institutions alike. This set of guidelines includes new sections on training, informed consent, and a core evidence base, which we hope you will find useful and informative. PMID- 16365342 TI - National variations in cardiac service provision: how united is our kingdom? PMID- 16365343 TI - Images in cardiology. Multislice cardiac computed tomographic images of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA). PMID- 16365344 TI - Images in cardiology. Arterial tortuosity syndrome in a newborn. PMID- 16365345 TI - Images in cardiology. An unexpected complication during right jugular vein cannulation in a young adult. PMID- 16365347 TI - Images in cardiology. Aneurysm arising from the left sinus of Valsalva and rupturing into the left ventricle: a rare entity. PMID- 16365348 TI - Images in cardiology. Coronary air embolism after removal of central venous catheter. PMID- 16365341 TI - JBS 2: Joint British Societies' guidelines on prevention of cardiovascular disease in clinical practice. PMID- 16365349 TI - Influence of arterial compliance on presence and extent of ischaemia during stress echocardiography. AB - OBJECTIVE: To seek an association between total arterial compliance (TAC) and the extent of ischaemia at stress echocardiography. DESIGN: Cohort study. SETTING: Regional cardiac centre. METHODS: 255 consecutive patients (147 men; mean (SD) age 58 (8)) presenting for stress echocardiography for clinical indications were studied. Wall motion score index (WMSI) was calculated and ischaemia was defined by an inducible or worsening wall motion abnormality. Peak WMSI was used to reflect the extent of dysfunction (ischaemia or scar), and DeltaWMSI was indicative of extent of ischaemia. TAC was assessed at rest by simultaneous radial applanation tonometry and pulsed wave Doppler in all patients. RESULTS: Ischaemia was identified by stress echocardiography in 65 patients (25%). TAC was similar in the groups with negative and positive echocardiograms (1.08 (0.41) v 1.17 (0.51) ml/mm Hg, not significant). However, the extent of dysfunction was associated with TAC independently of age, blood pressure, risk factors, and use of a beta blocker. Moreover, the extent of ischaemia was determined by TAC, risk factors, and use of a beta blocker. CONCLUSION: While traditional cardiovascular risk factors are strong predictors of ischaemia on stress echocardiography, TAC is an independent predictor of the extent of ischaemia. PMID- 16365350 TI - Images in cardiology. Angiographic appearance of "tumour blush" produced by a large right atrial myxoma. PMID- 16365351 TI - Images in cardiology. A contraindication for internal mammary to coronary bypass. PMID- 16365352 TI - Transoesophageal echocardiographic assessment of mitral valve commissural morphology predicts outcome after balloon mitral valvotomy. AB - OBJECTIVE: To investigate the value of transoesophageal echocardiography in the assessment of commissural morphology and prediction of outcome after balloon mitral valvotomy (BMV). DESIGN: Prospective study. SETTING: Tertiary cardiac referral centre. PATIENTS: 72 consecutive patients (mean age 61.3 years, range 38 89 years) referred for BMV. INTERVENTIONS: Transoesophageal echocardiography was performed immediately before BMV and the mitral commissures were scanned systematically. Anterolateral and posteromedial commissures were scored individually according to whether non-calcified fusion was absent (0), partial (1), or extensive (2). Calcified commissures usually resist splitting and scored 0. Scores for each commissure were combined giving an overall commissure score for each valve of 0-4, higher scores reflecting increased likelihood of commissural splitting. Valve anatomy was also graded by the method of Wilkins et al, which does not include commissural assessment. MAIN OUTCOME MEASURES: Patients were divided into outcome groups: A (good) and B (suboptimal). "Good" was defined as final valve area > 1.5 cm2 with a > 25% increase in area and absence of severe mitral regurgitation judged by echocardiography. RESULTS: Valve area increased from a mean (SD) of 1.1 (0.28) cm2 to 1.8 (0.46) cm2. Commissure scores were higher in group A than in group B (p < 0.01), scores > or = 2 predicting a good outcome with positive and negative accuracy of 67% and 82%, respectively (p < 0.001). Commissure score was the strongest independent predictor of outcome. CONCLUSION: Transoesophageal echocardiographic assessment of commissural morphology predicts outcome after BMV, adding significantly to the Wilkins score. PMID- 16365353 TI - Images in cardiology. Dizziness and dyspnoea: psychiatry and cardiology. PMID- 16365354 TI - Images in cardiology. Progressive Stanford B aortic dissection treated by acute stenting. PMID- 16365355 TI - Images in cardiology. Unusual neurological manifestation of acute mitral valve endocarditis. PMID- 16365356 TI - Images in cardiology. Percutaneous removal of embolised vegetation from left main coronary artery. PMID- 16365357 TI - Heart and heart-lung transplantation for idiopathic restrictive cardiomyopathy in children. AB - OBJECTIVE: To review the outcome of cardiac transplantation for restrictive cardiomyopathy (RCM) in children and to assess the ability of new strategies to modulate the effects of high pulmonary vascular resistance. DESIGN: Retrospective case note analysis of all patients receiving a transplant for RCM. PATIENTS: 18 children with RCM referred for transplantation assessment to Great Ormond Street Hospital, London. RESULTS: Eight boys and 10 girls were referred for assessment. Median age at presentation was 5.0 (mean (SD) 6.1 (4.0)) years. Fourteen orthotopic and two heterotopic transplantations were performed and two patients were referred for heart-lung transplantation. Mean duration from diagnosis to transplantation was 3.3 (3.0) years. Three patients with haemodynamic decompensation before transplantation had increased morbidity in the postoperative period. No patients died while awaiting a transplant. Three patients died in the first year after transplantation, one within 30 days. Five patients received pre-transplantation prostacyclin for a mean duration of 57 (18) days. Transpulmonary gradient was reduced in four of the patients. Mean transpulmonary gradient was 27 (9.8) mm Hg before and 17 (6.7) mm Hg after treatment with prostacyclin (p < 0.05). CONCLUSION: Most children with RCM require transplantation within four years of diagnosis. Referral for transplantation assessment should precede haemodynamic decompensation. Increase of pulmonary vascular resistance is a variable problem but can be modulated with pre-transplantation prostacyclin. With these strategies, orthotopic transplantation is possible in the majority of cases. PMID- 16365358 TI - Images in cardiology. Right coronary artery embolism from a thrombus of the ascending aorta. PMID- 16365359 TI - Images in cardiology. An unusual cause of chest pain and ST segment elevation. PMID- 16365360 TI - Images in cardiology. Massive pulmonary embolism without haemodynamic compromise caused by the presence of a patent foramen ovale. PMID- 16365361 TI - Role of reactive oxygen species on the formation of the novel diagnostic marker ischaemia modified albumin. PMID- 16365362 TI - Site of successful slow pathway ablation relates to clinical tachycardia rate in patients with atrioventricular nodal re-entrant tachycardia. PMID- 16365363 TI - Images in cardiology. Myocardial ischaemia in a patient with anomalous left coronary artery from the pulmonary artery. PMID- 16365364 TI - Dark chocolate improves endothelial and platelet function. PMID- 16365365 TI - N-terminal brain natriuretic peptide is predictive of death after cardiac transplantation. PMID- 16365366 TI - Images in cardiology. Acute recoil in sirolimus eluting stent: real time, in vivo assessment with optical coherence tomography. PMID- 16365367 TI - Management of infective endocarditis. PMID- 16365369 TI - Role of real time 3D echocardiography in evaluating the left ventricle. PMID- 16365370 TI - Aortic dimensions and the risk of dissection. PMID- 16365371 TI - Magnetic resonance imaging of articular cartilage: trauma, degeneration, and repair. AB - The assessment of articular cartilage using magnetic resonance imaging has seen considerable advances in recent years. Cartilage morphologic characteristics can now be evaluated with a high degree of accuracy and reproducibility using dedicated pulse sequences, which are becoming standard at many institutions. These techniques detect clinically unsuspected traumatic cartilage lesions, allowing the physician to study their natural history with longitudinal evaluation and also to assess disease status in degenerative osteoarthritis. Magnetic resonance imaging also provides a more objective assessment of cartilage repair to augment the information obtained from more subjective clinical outcome instruments. Newly developed methods that provide detail at an ultrastructural level offer an important addition to cartilage evaluation, particularly in the detection of early alterations in the extracellular matrix. These methods have created an undeniably important role for magnetic resonance imaging in the reproducible, noninvasive, and objective evaluation and monitoring of cartilage. An overview of the advances, current techniques, and impact of magnetic resonance imaging in the setting of trauma, degenerative arthritides, and surgical treatment for cartilage injury is presented. PMID- 16365372 TI - Arthroscopic treatment of posterolateral elbow impingement from lateral synovial plicae in throwing athletes and golfers. AB - BACKGROUND: Although elbow pain is common in throwing athletes and golfers, posterolateral impingement from a hypertrophic synovial plica is a rare but possibly underdiagnosed condition. PURPOSE: To evaluate the clinical results of arthroscopic treatment of symptomatic lateral elbow plicae in this athletic population. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Twelve patients, 9 male and 3 female, whose mean age was 21.6 years (range, 17-33 years), were reviewed. There were 7 baseball pitchers, 2 softball players, and 3 golfers. All patients had diagnosed isolated lateral elbow plica; none had lateral epicondylitis, instability, osteochondritis dissecans, arthritis, loose bodies, or nerve conditions. The mean time from onset of symptoms to treatment was 9.25 months (range, 3-24.5 months). At a mean follow-up of 33.8 months (range, 24-65.5 months), patients were evaluated with a questionnaire and examination, based on the American Shoulder and Elbow Surgeons standardized elbow assessment and previously reported elbow outcome score. RESULTS: Posterolateral elbow pain was present in all patients. Fifty-eight percent (7 of 12 patients) complained of clicking or catching, whereas only 25% (3 of 12) experienced swelling or effusion. At elbow arthroscopy, a thickened synovial lateral plica was debrided in all cases. Ninety-two percent (11 of 12 patients) reported an excellent outcome with a mean elbow score of 92.5 points (maximum, 100 points). Return to competitive play averaged 4.8 months (range, 3-9.5 months). One patient with a fair outcome developed medial elbow instability that later required reconstructive surgery. CONCLUSION: Posterolateral elbow impingement from hypertrophic synovial plicae can cause significant pain and disability in throwing athletes and golfers. With careful diagnosis and exclusion of other elbow problems, treatment with arthroscopic debridement and focused rehabilitation is highly successful and allows these athletes to return to their previous level of play. PMID- 16365373 TI - Anterior cruciate ligament revision reconstruction: results using a quadriceps tendon-patellar bone autograft. AB - BACKGROUND: The quadriceps tendon is a viable graft source for revision anterior cruciate ligament reconstruction. PURPOSE: To determine the functional results and graft failure rates in knees in which the patellar tendon had been previously harvested or was unavailable, expanded tunnels precluded the use of a semitendinosus-gracilis graft, or patients requested autogenous tissues instead of allografts for revision reconstruction. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The authors observed 21 patients for a mean of 49 months postoperatively after revision anterior cruciate ligament reconstruction with a quadriceps tendon graft. The results were determined by KT-2000 arthrometer testing, the Cincinnati Knee Rating System, and the International Knee Documentation Committee Rating System. Fifteen knees required a concurrent procedure, including reconstruction of posterolateral structures in 5 knees, meniscal repairs in 5 knees, and high tibial osteotomy in 2 knees. RESULTS: Significant improvements occurred in symptoms (P < .0001), daily activities (P < .05), sports activities (P < .01), and the overall rating scores (P < .0001). Eighteen patients rated their knee condition as improved. Total mean anterior posterior displacements decreased from 8.4 +/- 3.1 mm preoperatively to 2.0 +/- 2.3 mm at follow-up (P < .001). On the International Knee Documentation Committee knee ligament rating, 17 knees were graded as normal or nearly normal, 3 were graded as abnormal, and 1 was graded as severely abnormal. CONCLUSION: The revision operation provided reasonable results in this group of complex knees. However, the functional and overall results were inferior to those reported for primary anterior cruciate ligament reconstruction. Many knees (90%) had compounding problems of articular cartilage damage, meniscectomy, varus malalignment, or additional ligamentous injury that most likely affected the results. PMID- 16365374 TI - Where should the femoral tunnel of a posterior cruciate ligament reconstruction be placed to best restore anteroposterior laxity and ligament forces? AB - BACKGROUND: Objective results of posterior cruciate ligament reconstruction are often less than satisfactory, with many patients exhibiting excessive posterior laxity. HYPOTHESIS: Changes in the position of the femoral tunnel within the posterior cruciate ligament's femoral footprint will significantly affect anteroposterior laxities and graft forces. STUDY DESIGN: Controlled laboratory study. METHODS: The posterior cruciate ligament's femoral origin was mechanically isolated in 13 fresh-frozen knee specimens, and the bone cap containing the ligament's insertion was attached to a load cell that recorded resultant force during tibial loading tests. Anteroposterior laxity (at +/- 200 N applied force) was also measured. Cast acrylic replicas of the bone cap were fabricated, with tunnels placed in anterolateral, central, and posteromedial regions of the footprint. A graft reconstruction was tested in each tunnel. RESULTS: Mean laxities with the anterolateral tunnel were +0.9 mm to +1.7 mm greater than normal between 0 degrees and 45 degrees of flexion. Mean laxities with the posteromedial tunnel were -2.4 mm to -3.7 mm less than normal between 10 degrees and 45 degrees of flexion. Mean laxities with the central tunnel were not significantly different from intact knee values, except at 0 degrees (0.9 mm greater). Mean graft forces with the anterolateral tunnel were normal for most modes of loading, whereas there were significant increases in graft forces with the posteromedial and central tunnels. CONCLUSION: The anterolateral tunnel reproduced normal posterior cruciate ligament force profiles but produced a knee that was more lax than normal between 0 degrees and 45 degrees of flexion. The central tunnel best matched intact knee laxities, but graft forces were higher than posterior cruciate ligament forces between 0 degrees and 45 degrees of flexion. The posteromedial tunnel overconstrained anteroposterior laxity approximately 2 to 4 mm between 0 degrees and 45 degrees of flexion and generated higher graft forces in the same flexion range. CLINICAL RELEVANCE: This study suggests that a posteromedial tunnel should not be used for single-bundle posterior cruciate ligament reconstruction. PMID- 16365375 TI - The role of hip muscle function in the treatment of patellofemoral pain syndrome. AB - BACKGROUND: Previous literature has associated hip weakness with patellofemoral pain syndrome. HYPOTHESIS: Improvements in hip strength and flexibility are associated with a decrease in patellofemoral pain. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Thirty-five patients with patellofemoral pain syndrome, aged 33 +mn; 16 years (29 women, 6 men; 43 knees), were evaluated and placed on a 6-week treatment program. Hip flexion, abduction, and adduction strengths, Thomas and Ober test results, and visual analog scale scores for pain with activities of daily living as well as with exercise were documented on initial evaluation and again 6 weeks later. Treatment consisted of strength and flexibility exercises primarily focusing on the hip. RESULTS: Hip flexion strength improved by 35% +/- 8.4% in 26 lower extremities treated successfully, compared with -1.8% +/- 3.5% in 17 lower extremities with an unsuccessful outcome (P < .001). Before treatment, there were positive Ober test results in 39 of 43 lower extremities; positive Thomas test results were seen in 31 of 43 lower extremities. A successful outcome with a concurrent normalized Ober test result was seen in 83% (20/24) of lower extremities, and successful outcomes with normalized Thomas test results were seen in 80% (16/20) of lower extremities. A combination of improved hip flexion strength (> 20%) as well as normal Ober and Thomas test results was seen in 93% of successfully treated cases (14/15 lower extremities), compared with 0% success (0/5 lower extremities) if there was no change in hip flexion strength (< 20%) and if Ober and Thomas test results remained positive. CONCLUSIONS: Improvements in hip flexion strength combined with increased iliotibial band and iliopsoas flexibility were associated with excellent results in patients with patellofemoral pain syndrome. PMID- 16365376 TI - The influence of intraoperative pretensioning on the chondroprotective effect of meniscal transplants. AB - BACKGROUND: Meniscal replacement has become more common in recent years. The meniscal transplant's ability to transfer load effectively depends on its ability to bear circumferential loading. HYPOTHESIS: Intraoperative pretensioning on the meniscal transplant sutures has a positive influence on meniscal transplants' chondroprotective effect. STUDY DESIGN: Controlled laboratory study. METHODS: Thirty-six sheep were divided into 6 groups (n = 6), subjected to a sham operation (group A), a meniscectomy (group B), or a meniscal autograft using tag sutures with different levels of pretensioning (group C, 0 N; group D, 20 N; group E, 40 N; group F, 60 N). Macroscopic (International Cartilage Repair Society score) and histologic evaluations (Mankin score) of the articular cartilage were performed after 6 months. RESULTS: Higher suture pretension (40 N, 60 N) resulted in less cartilage degeneration than in meniscectomized (P =.047; P =.036) and nonpretensioned (P =.028; P =.015) knees, with International Cartilage Repair Society scores of 1.63 +/- 0.57 and 1.66 +/- 0.51 in groups E and F, respectively, and scores of 2.40 +/- 0.27 and 2.68 +/- 0.46 observed after meniscectomy and meniscal transplantation with no pretensioning, respectively. Group F had a significantly better Mankin score of 6.66 +/- 2.15 (P =.05) compared with group D. Regarding criterion cells, trends toward less degeneration compared with meniscectomized and nonpretensioned knees (P = .054 and P =.055) were found. The coefficient of variation of the Mankin scores was greater than that of the International Cartilage Repair Society score. Group A had significantly better cartilage than all other groups. CONCLUSION: Adequate intraoperative pretensioning has a significant influence on the chondroprotective effect of meniscal transplants but did not prevent the development of articular cartilage degeneration. CLINICAL RELEVANCE: The results suggest that intraoperative pretensioning could improve the chondroprotective effect of meniscal transplantation. PMID- 16365377 TI - Massive genome erosion and functional adaptations provide insights into the symbiotic lifestyle of Sodalis glossinidius in the tsetse host. AB - Sodalis glossinidius is a maternally transmitted endosymbiont of tsetse flies (Glossina spp.), an insect of medical and veterinary significance. Analysis of the complete sequence of Sodalis' chromosome (4,171,146 bp, encoding 2,432 protein coding sequences) indicates a reduced coding capacity of 51%. Furthermore, the chromosome contains 972 pseudogenes, an inordinately high number compared with that of other bacterial species. A high proportion of these pseudogenes are homologs of known proteins that function either in defense or in the transport and metabolism of carbohydrates and inorganic ions, suggesting Sodalis' degenerative adaptations to the immunity and restricted nutritional status of the host. Sodalis possesses three chromosomal symbiosis regions (SSR): SSR-1, SSR-2, and SSR-3, with gene inventories similar to the Type-III secretion system (TTSS) ysa from Yersinia enterolitica and SPI-1 and SPI-2 from Salmonella, respectively. While core components of the needle structure have been conserved, some of the effectors and regulators typically associated with these systems in pathogenic microbes are modified or eliminated in Sodalis. Analysis of SSR specific invA transcript abundance in Sodalis during host development indicates that the individual symbiosis regions may exhibit different temporal expression profiles. In addition, the Sodalis chromosome encodes a complete flagella structure, key components of which are expressed in immature host developmental stages. These features may be important for the transmission and establishment of symbiont infections in the intra-uterine progeny. The data suggest that Sodalis represents an evolutionary intermediate transitioning from a free-living to a mutualistic lifestyle. PMID- 16365379 TI - Evolution of alternative splicing after gene duplication. AB - Alternative splicing and gene duplication are two major sources of proteomic function diversity. Here, we study the evolutionary trend of alternative splicing after gene duplication by analyzing the alternative splicing differences between duplicate genes. We observed that duplicate genes have fewer alternative splice (AS) forms than single-copy genes, and that a negative correlation exists between the mean number of AS forms and the gene family size. Interestingly, we found that the loss of alternative splicing in duplicate genes may occur shortly after the gene duplication. These results support the subfunctionization model of alternative splicing in the early stage after gene duplication. Further analysis of the alternative splicing distribution in human duplicate pairs showed the asymmetric evolution of alternative splicing after gene duplications; i.e., the AS forms between duplicates may differ dramatically. We therefore conclude that alternative splicing and gene duplication may not evolve independently. In the early stage after gene duplication, young duplicates may take over a certain amount of protein function diversity that previously was carried out by the alternative splicing mechanism. In the late stage, the gain and loss of alternative splicing seem to be independent between duplicates. PMID- 16365378 TI - Defining the mammalian CArGome. AB - Serum response factor (SRF) binds a 1216-fold degenerate cis element known as the CArG box. CArG boxes are found primarily in muscle- and growth-factor-associated genes although the full spectrum of functional CArG elements in the genome (the CArGome) has yet to be defined. Here we describe a genome-wide screen to further define the functional mammalian CArGome. A computational approach involving comparative genomic analyses of human and mouse orthologous genes uncovered >100 hypothetical SRF-dependent genes, including 10 previously identified SRF targets, harboring a conserved CArG element within 4000 bp of the annotated transcription start site (TSS). We PCR-cloned 89 hypothetical SRF targets and subjected each of them to at least two of several validations including luciferase reporter, gel shift, chromatin immunoprecipitation, and mRNA expression following RNAi knockdown of SRF; 60/89 (67%) of the targets were validated. Interestingly, 26 of the validated SRF target genes encode for cytoskeletal/contractile or adhesion proteins. RNAi knockdown of SRF diminishes expression of several SRF-dependent cytoskeletal genes and elicits an attending perturbation in the cytoarchitecture of both human and rodent cells. These data illustrate the power of integrating existing algorithms to interrogate the genome in a relatively unbiased fashion for cis-regulatory element discovery. In this manner, we have further expanded the mammalian CArGome with the discovery of an array of cyto-contractile genes that coordinate normal cytoskeletal homeostasis. We suggest one function of SRF is that of an ancient master regulator of the actin cytoskeleton. PMID- 16365380 TI - A genome-wide study of dual coding regions in human alternatively spliced genes. AB - Alternative splicing is a major mechanism for gene product regulation in many multicellular organisms. By using different exon combinations, some coding regions can encode amino acids in multiple reading frames in different transcripts. Here we performed a systematic search through a set of high-quality human transcripts and show that approximately 7% of alternatively spliced genes contain dual (multiple) coding regions. By using a conservative criterion, we found that in these regions most secondary reading frames evolved recently in mammals, and a significant proportion of them may be specific to primates. Based on the presence of in-frame stop codons in orthologous sequences in other animals, we further classified ancestral and derived reading frames in these regions. Our results indicated that ancestral reading frames are usually under stronger selection than are derived reading frames. Ancestral reading frames mainly influence the coding properties of these dual coding regions. Compared with coding regions of the whole genome, ancestral reading frames largely maintain similar nucleotide composition at each codon position and amino acid usage, while derived reading frames are significantly different. Our results also indicated that prior to acquisition of a new reading frame, the suppression of in frame stop codons in the ancestral state is mainly achieved by one-step transition substitutions at the first or second codon position. Finally, the selective forces imposed on these dual coding regions will also be discussed. PMID- 16365381 TI - Large-scale structure of genomic methylation patterns. AB - The mammalian genome depends on patterns of methylated cytosines for normal function, but the relationship between genomic methylation patterns and the underlying sequence is unclear. We have characterized the methylation landscape of the human genome by global analysis of patterns of CpG depletion and by direct sequencing of 3073 unmethylated domains and 2565 methylated domains from human brain DNA. The genome was found to consist of short (<4 kb) unmethylated domains embedded in a matrix of long methylated domains. Unmethylated domains were enriched in promoters, CpG islands, and first exons, while methylated domains comprised interspersed and tandem-repeated sequences, exons other than first exons, and non-annotated single-copy sequences that are depleted in the CpG dinucleotide. The enrichment of regulatory sequences in the relatively small unmethylated compartment suggests that cytosine methylation constrains the effective size of the genome through the selective exposure of regulatory sequences. This buffers regulatory networks against changes in total genome size and provides an explanation for the C value paradox, which concerns the wide variations in genome size that scale independently of gene number. This suggestion is compatible with the finding that cytosine methylation is universal among large-genome eukaryotes, while many eukaryotes with genome sizes <5 x 10(8) bp do not methylate their DNA. PMID- 16365382 TI - Design optimization methods for genomic DNA tiling arrays. AB - A recent development in microarray research entails the unbiased coverage, or tiling, of genomic DNA for the large-scale identification of transcribed sequences and regulatory elements. A central issue in designing tiling arrays is that of arriving at a single-copy tile path, as significant sequence cross hybridization can result from the presence of non-unique probes on the array. Due to the fragmentation of genomic DNA caused by the widespread distribution of repetitive elements, the problem of obtaining adequate sequence coverage increases with the sizes of subsequence tiles that are to be included in the design. This becomes increasingly problematic when considering complex eukaryotic genomes that contain many thousands of interspersed repeats. The general problem of sequence tiling can be framed as finding an optimal partitioning of non repetitive subsequences over a prescribed range of tile sizes, on a DNA sequence comprising repetitive and non-repetitive regions. Exact solutions to the tiling problem become computationally infeasible when applied to large genomes, but successive optimizations are developed that allow their practical implementation. These include an efficient method for determining the degree of similarity of many oligonucleotide sequences over large genomes, and two algorithms for finding an optimal tile path composed of longer sequence tiles. The first algorithm, a dynamic programming approach, finds an optimal tiling in linear time and space; the second applies a heuristic search to reduce the space complexity to a constant requirement. A Web resource has also been developed, accessible at http://tiling.gersteinlab.org, to generate optimal tile paths from user-provided DNA sequences. PMID- 16365383 TI - Identification by full-coverage array CGH of human DNA copy number increases relative to chimpanzee and gorilla. AB - Duplication of chromosomal segments and associated genes is thought to be a primary mechanism for generating evolutionary novelty. By comparative genome hybridization using a full-coverage (tiling) human BAC array with 79-kb resolution, we have identified 63 chromosomal segments, ranging in size from 0.65 to 1.3 Mb, that have inferred copy number increases in human relative to chimpanzee. These segments span 192 Ensembl genes, including 82 gene duplicates (41 reciprocal best BLAST matches). Synonymous and nonsynonymous substitution rates across these pairs provide evidence for general conservation of the amino acid sequence, consistent with the maintenance of function of both copies, and one case of putative positive selection for an uncharacterized gene. Surprisingly, the core histone genes H2A, H2B, H3, and H4 have been duplicated in the human lineage since our split with chimpanzee. The observation of increased copy number of a human cluster of core histone genes suggests that altered dosage, even of highly constrained genes, may be an important evolutionary mechanism. PMID- 16365384 TI - L1 integration in a transgenic mouse model. AB - To study integration of the human LINE-1 retrotransposon (L1) in vivo, we developed a transgenic mouse model of L1 retrotransposition that displays de novo somatic L1 insertions at a high frequency, occasionally several insertions per mouse. We mapped 3' integration sites of 51 insertions by Thermal Asymmetric Interlaced PCR (TAIL-PCR). Analysis of integration locations revealed a broad genomic distribution with a modest preference for intergenic regions. We characterized the complete structures of 33 de novo retrotransposition events. Our results highlight the large number of highly truncated L1s, as over 52% (27/51) of total integrants were <1/3 the length of a full-length element. New integrants carry all structural characteristics typical of genomic L1s, including a number with inversions, deletions, and 5'-end microhomologies to the target DNA sequence. Notably, at least 13% (7/51) of all insertions contain a short stretch of extra nucleotides at their 5' end, which we postulate result from template jumping by the L1-encoded reverse transcriptase. We propose a unified model of L1 integration that explains all of the characteristic features of L1 retrotransposition, such as 5' truncations, inversions, extra nucleotide additions, and 5' boundary and inversion point microhomologies. PMID- 16365385 TI - Transposon-free regions in mammalian genomes. AB - Despite the presence of over 3 million transposons separated on average by approximately 500 bp, the human and mouse genomes each contain almost 1000 transposon-free regions (TFRs) over 10 kb in length. The majority of human TFRs correlate with orthologous TFRs in the mouse, despite the fact that most transposons are lineage specific. Many human TFRs also overlap with orthologous TFRs in the marsupial opossum, indicating that these regions have remained refractory to transposon insertion for long evolutionary periods. Over 90% of the bases covered by TFRs are noncoding, much of which is not highly conserved. Most TFRs are not associated with unusual nucleotide composition, but are significantly associated with genes encoding developmental regulators, suggesting that they represent extended regions of regulatory information that are largely unable to tolerate insertions, a conclusion difficult to reconcile with current conceptions of gene regulation. PMID- 16365386 TI - Overexpression of phosphodiesterases in experimental autoimmune myasthenia gravis: suppression of disease by a phosphodiesterase inhibitor. AB - Myasthenia gravis (MG) and experimental autoimmune MG (EAMG) are T cell-dependent antibody-mediated autoimmune disorders, in which the nicotinic acetylcholine receptor (AChR) is the major autoantigen. DNA microarray analysis revealed increased levels of several phosphodiesterase (PDE) subtypes in lymph node cells (LNC) and muscles of EAMG rats compared with healthy controls. Quantitative real time PCR analysis indicated that EAMG is characterized by an increase of PDE subtypes 1, 3, 4, and 7 in LNC and of PDE subtypes 2, 3, 4, and 7 in muscles. Pentoxifylline (PTX), a general PDE inhibitor, inhibited the progression of EAMG when treatment started at either the acute or chronic stages of disease. This suppression was associated with down-regulation of humoral and cellular AChR specific responses, as well as down-regulation of PDE4, TNF-alpha, IL-18, IL-12, and IL-10 in LNC and of PDEs 1, 4, 7, and TNF-alpha in muscles. The expression of Foxp3, a transcription factor essential for CD4+CD25+ regulatory T cell function, was increased in splenocytes although the number of these cells remained unchanged. PTX also reduced the expression of the endopeptidase cathepsin-l, a marker of muscle damage, in EAMG muscles. This study demonstrates the involvement of PDE regulation in EAMG pathogenesis and suggests that PDE inhibitors may be considered for immunotherapy of MG. PMID- 16365387 TI - Shaping the T cell repertoire. PMID- 16365388 TI - In a radiation chimaera, host H-2 antigens determine immune responsiveness of donor cytotoxic cells. Nature 1977. 269: 417-418. PMID- 16365389 TI - Cutting edge: the AP-1 subunit JunB determines NK cell-mediated target cell killing by regulation of the NKG2D-ligand RAE-1epsilon. AB - The activating receptor NKG2D and its ligands RAE-1 play an important role in the NK, gammadelta+, and CD8+ T cell-mediated immune response to tumors. Expression levels of RAE-1 on target cells have to be tightly controlled to allow immune cell activation against tumors but to avoid destruction of healthy tissues. In this study, we report that cell surface expression of RAE-1epsilon is greatly enhanced on cells lacking JunB, a subunit of the transcription complex AP-1. Furthermore, tissue-specific junB knockout mice respond to 12-O-tetradecanoyl phorbol-13-acetate, a potent AP-1 activator, with markedly increased and sustained epidermal RAE-1epsilon expression. Accordingly, junB-deficient cells are efficiently killed via NKG2D by NK cells and induce IFN-gamma production. Our data indicate that the transcription factor AP-1, which is involved in tumorigenesis and cellular stress responses, regulates RAE-1epsilon. Thus, up regulated RAE-1epsilon expression due to low levels of JunB could alert immune cells to tumors and stressed cells. PMID- 16365390 TI - Cutting edge: extracellular high mobility group box-1 protein is a proangiogenic cytokine. AB - The chromosomal high mobility group box-1 (HMGB1) protein acts as a proinflammatory cytokine when released in the extracellular environment by necrotic and inflammatory cells. In the present study, we show that HMGB1 exerts proangiogenic effects by inducing MAPK ERK1/2 activation, cell proliferation, and chemotaxis in endothelial cells of different origin. Accordingly, HMGB1 stimulates membrane ruffling and repair of a mechanically wounded endothelial cell monolayer and causes endothelial cell sprouting in a three-dimensional fibrin gel. In keeping with its in vitro properties, HMGB1 stimulates neovascularization when applied in vivo on the top of the chicken embryo chorioallantoic membrane whose blood vessels express the HMGB1 receptor for advanced glycation end products (RAGE). Accordingly, RAGE blockade by neutralizing Abs inhibits HMGB1-induced neovascularization in vivo and endothelial cell proliferation and membrane ruffling in vitro. Taken together, the data identify HMGB1/RAGE interaction as a potent proangiogenic stimulus. PMID- 16365391 TI - A key regulatory role for histamine in experimental autoimmune encephalomyelitis: disease exacerbation in histidine decarboxylase-deficient mice. AB - Histamine can modulate the cytokine network and influence Th1 and Th2 balance and Ab-isotype switching. Thus, pharmacological blockade or genetic deletion of specific histamine receptors has been shown to reduce the severity of experimental autoimmune encephalomyelitis (EAE), a prototypic Th1-mediated disease with similarities to human multiple sclerosis. To study the comprehensive contribution of endogenous histamine to the expression of EAE, we attempted to induce EAE in histidine decarboxylase-deficient mice, which are genetically unable to make histamine. In this study, we show that EAE is significantly more severe in HDC-/-, histamine-deficient mice, with diffuse inflammatory infiltrates, including a prevalent granulocytic component, in the brain and cerebellum. Unlike splenocytes from wild-type mice, splenocytes from HDC-/- mice do not produce histamine in response to the myelin Ag, whereas production of IFN gamma, TNF, and leptin are increased in HDC-/- splenocytes in comparison to those from wild-type mice. Endogenous histamine thus appears to regulate importantly the autoimmune response against myelin and the expression of EAE, in this model, and to limit immune damage to the CNS. Understanding which receptor(s) for histamine is/are involved in regulating autoimmunity against the CNS might help in the development of new strategies of treatment for EAE and multiple sclerosis. PMID- 16365392 TI - Hydrolytic and nonenzymatic functions of acetylcholinesterase comodulate hemopoietic stress responses. AB - Glucocorticoid-initiated granulocytosis, excessive proliferation of granulocytes, persists after cortisol levels are lowered, suggesting the involvement of additional stress mediator(s). In this study, we report that the stress-induced acetylcholinesterase variant, AChE-R, and its cleavable, cell-penetrating C terminal peptide, ARP, facilitate granulocytosis. In postdelivery patients, AChE R-expressing granulocyte counts increased concomitantly with serum cortisol and AChE activity levels, yet persisted after cortisol had declined. Ex vivo, mononuclear cells of adult peripheral blood responded to synthetic ARP26 by overproduction of hemopoietically active proinflammatory cytokines (e.g., IL-6, IL-10, and TNF-alpha). Physiologically relevant ARP26)levels promoted AChE gene expression and induced the expansion of cultured CD34+ progenitors and granulocyte maturation more effectively than cortisol, suggesting autoregulatory prolongation of ARP effects. In vivo, transgenic mice overexpressing human AChE R, unlike matched controls, showed enhanced expression of the myelopoietic transcription factor PU.1 and maintained a stable granulocytic state following bacterial LPS exposure. AChE-R accumulation and the consequent inflammatory consequences can thus modulate immune responses to stress stimuli. PMID- 16365393 TI - Novel insights into the mechanism of action of FTY720 in a transgenic model of allograft rejection: implications for therapy of chronic rejection. AB - FTY720 is a high-affinity agonist at the sphingosine 1-phosphate receptor 1 that prevents lymphocyte egress from lymphoid tissue and prolongs allograft survival in several animal models of solid organ transplantation. In this study we used a recently developed adoptive transfer model of TCR transgenic T cells to track allospecific CD4+ T cell expansion and trafficking characteristics, cytokine secretion profiles, and surface phenotype in vivo in the setting of FTY720 administration. We report that FTY720 administration had no effect on alloantigen driven T cell activation, proliferation, acquisition of effector-memory function, or T cell apoptosis. However, FTY720 caused a reversible sequestration of alloantigen-specific effector-memory T cells in regional lymphoid tissue associated with a decrease in T cell infiltration within the allograft and a subsequent prolongation in allograft survival. Furthermore, delayed administration of FTY720 in a cardiac model of chronic allograft rejection attenuated the progression of vasculopathy and tissue fibrosis consistent with the hypothesis that FTY720 interrupts the trafficking of activated effector memory T cells. These data have important implications for targeting the sphingosine 1-phosphate receptor 1 in solid organ transplantation. PMID- 16365394 TI - The membrane-bound chemokine CXCL16 expressed on follicle-associated epithelium and M cells mediates lympho-epithelial interaction in GALT. AB - The recently identified CXCL16 has dual functions as a transmembrane adhesion molecule and a soluble chemokine. In this study we found that CXCL16 mRNA and protein were expressed constitutively on the follicle-associated epithelium covering Peyer's patches (PPs), isolated lymphoid follicles, and cecal patches, but minimally on the villous epithelium in the murine gastrointestinal tract. The CXCL16 receptor CXCR6/Bonzo was constitutively expressed on subpopulations of CD4+ and CD8+ T cells isolated from PPs. The expression of CXCR6/Bonzo on the PP T cells was up-regulated after stimulation with anti-CD3 and anti-CD28 mAbs. The activated PP T cells showed chemotactic migration in response to the soluble N terminal chemokine domain of CXCL16. Furthermore, the activated PP T cells selectively adhered to cells expressing murine CXCL16. To determine the physiological role of CXCL16 in GALT, we first carefully analyzed T cell distribution in PPs. T cells localized not only in the interfollicular region but also at a lesser frequency in the subepithelial dome (SED) and in the germinal center of lymphoid follicles. Consistently, the majority of the adoptive transferred activated T cells migrated into the SED and the interfollicular region. However, the neutralization of CXCL16 specifically reduced the migration of the adoptive, transferred, activated T cells into the SED of PPs. These data suggest that CXCL16 expressed on the follicle-associated epithelium plays an important role in the recruitment and retention of activated T cells in the SED and should, at least partially, be responsible for lymphocyte compartmentalization in GALT. PMID- 16365395 TI - Integrin alphaIIbbeta3 induces the adhesion and activation of mast cells through interaction with fibrinogen. AB - Integrin alphaIIb, a well-known marker of megakaryocyte-platelet lineage, has been recently recognized on hemopoietic progenitors. We now demonstrate that integrin alphaIIbbeta3 is highly expressed on mouse and human mast cells including mouse bone marrow-derived mast cells, peritoneal mast cells, and human cord blood-derived mast cells, and that its binding to extracellular matrix proteins leads to enhancement of biological functions of mast cells in concert with various stimuli. With exposure to various stimuli, including cross-linking of FcepsilonRI and stem cell factor, mast cells adhered to extracellular matrix proteins such as fibrinogen and von Willebrand factor in an integrin alphaIIbbeta3-dependent manner. In addition, the binding of mast cells to fibrinogen enhanced proliferation, cytokine production, and migration and induced uptake of soluble fibrinogen in response to stem cell factor stimulation, implicating integrin alphaIIbbeta3 in a variety of mast cell functions. In conclusion, mouse and human mast cells express functional integrin alphaIIbbeta3. PMID- 16365396 TI - Tumor-infiltrating dendritic cells are potent antigen-presenting cells able to activate T cells and mediate tumor rejection. AB - Dendritic cells (DC) are potent inducers of immune responses. DC have been shown to infiltrate tumors, but very little is known about the functional status of these naturally occurring tumor-infiltrating DC (TIDC). In this study, the status and function of TIDC from several types of mouse melanoma were investigated in detail. CD11c+/MHC II+ cells, consistent with a DC phenotype, were found in all of transplantable or spontaneous melanomas studied. These TIDC were predominantly myeloid (CD11c+/CD8alpha-/B220-) in nature with small numbers of plasmacytoid (CD11c+/B220+). TIDC had an intermediate maturation phenotype with some expression of costimulatory molecules and the capacity to take up particles. Upon culture overnight ex vivo, the TIDC markedly up-regulated the expression of costimulatory molecules and also increased IL-12 production. Importantly, such ex vivo-matured TIDC pulsed with OVA were able to migrate to lymph nodes, to activate naive OVA-specific CD4+ and CD8+ T cells, and to confer protection against a challenge with OVA-expressing tumor cells. In conclusion, melanomas are infiltrated by functional DC that can act as fully competent APC. These APC have the potential to be manipulated and may therefore represent a promising target for cancer immunotherapy. PMID- 16365398 TI - Premature expression of chemokine receptor CCR9 impairs T cell development. AB - During thymocyte development, CCR9 is expressed on late CD4-CD8- (double-negative (DN)) and CD4+CD8+ (double-positive) cells, but is subsequently down-regulated as cells transition to the mature CD4+ or CD8+ (single-positive (SP)) stage. This pattern of expression has led to speculation that CCR9 may regulate thymocyte trafficking and/or export. In this study, we generated transgenic mice in which CCR9 surface expression was maintained throughout T cell development. Significantly, forced expression of CCR9 on mature SP thymocytes did not inhibit their export from the thymus, indicating that CCR9 down-regulation is not essential for thymocyte emigration. CCR9 was also expressed prematurely on immature DN thymocytes in CCR9 transgenic mice. Early expression of CCR9 resulted in a partial block of development at the DN stage and a marked reduction in the numbers of double-positive and SP thymocytes. Moreover, in CCR9-transgenic mice, CD25high DN cells were scattered throughout the cortex rather than confined to the subcapsular region of the thymus. Together, these results suggest that regulated expression of CCR9 is critical for normal development of immature thymocytes, but that down-regulation of CCR9 is not a prerequisite for thymocyte emigration. PMID- 16365397 TI - Heterogeneous nuclear ribonucleoprotein P2 is an autoantibody target in mice deficient for Mer, Axl, and Tyro3 receptor tyrosine kinases. AB - Deficiencies in clearance of apoptotic cells predispose to the development of autoimmune disease. This is evident in mice lacking the receptor tyrosine kinases Tyro3, Axl, and Mer. Deficient mice exhibit an increased abundance of apoptotic cells in tissues and manifest diverse autoimmune conditions. To test these mice for the presence of autoantibodies to apoptotic cells, we generated spontaneous splenic B cell hybridomas and used a novel microscopy screen to detect Ab binding to apoptotic Jurkat cells. From hybridomas secreting IgG Abs reactive with apoptotic cells, we selected one that recreated the major serum specificity for apoptotic cells. The Ab LHC7.15 bound to an Ag that is differentially distributed between the nucleus and the cytoplasm in live and apoptotic cells. In late apoptotic cells, the Ag coalesces into aggregates that bleb from the cell surface. Immunopurification of the Ag, followed by mass spectrometry, identified a protein of 69 kDa whose partial sequence matched heterogeneous nuclear ribonucleoprotein P2. This multifunctional protein binds DNA, RNA, and several known ribonucleoprotein autoantigens. Our observations indicate that a ribonucleoprotein complex, formed and translocated to the cell surface in apoptosis, represents a potent stimulus for breaking tolerance and inducing systemic autoimmunity in mice with defective clearance of cell remnants. PMID- 16365399 TI - Improved vaccine protection from simian AIDS by the addition of nonstructural simian immunodeficiency virus genes. AB - An HIV-1 vaccine able to induce broad CD4+ and CD8+ T cell responses may provide long-term control of viral replication. In this study we directly assess the relative benefit of immunization with vaccines expressing three structural Ags (Gag, Pol, and Env), three early regulatory proteins (Rev, Tat, and Nef), or a complex vaccine expressing all six Ags. The simultaneous administration of all six Ags during vaccination resulted in Ag competition manifested by a relative reduction of CD8+ T cell and lymphoproliferative responses to individual Ags. Despite the Ag competition, vaccination with all six Ags resulted in a delay in the onset and a decrease in the extent of acute viremia after mucosal challenge exposure to highly pathogenic SIV(mac251). Reduced levels of acute viremia correlated with lower post-set point viremia and long-term control of infection. In immunized animals, virus-specific CD4+ T cell and lymphoproliferative responses were preserved during acute viremia, and the maintenance of these responses predicted the long-term virological outcome. Taken together, these results suggest that the breadth of the immune response is probably more important than high frequency responses to a limited number of epitopes. These data provide the first clear evidence of the importance of nonstructural HIV Ags as components of an HIV-1 vaccine. PMID- 16365400 TI - Granzyme B, a new player in activation-induced cell death, is down-regulated by vasoactive intestinal peptide in Th2 but not Th1 effectors. AB - Following antigenic stimulation and differentiation, Th1 and Th2 effector cells contribute differently to cellular and humoral immunity. Vasoactive intestinal peptide (VIP) induces Th2 responses by promoting Th2 differentiation and survival. In this study, we investigate the mechanisms for the protective effect of VIP against activation-induced cell death (AICD) of Th2 effectors. Surprisingly, microarray and protein data indicate that VIP prevents the up regulation of granzyme B (GrB) in Th2 but not Th1 effectors. This is the first report of GrB expression in Th cells and of its involvement in activation-induced apoptosis. The enhanced responsiveness of Th2 cells to VIP is probably due to the higher expression of VIP receptors. The effect of VIP on Th2 survival and GrB expression is mediated through the VIP receptors 1 and 2 and cAMP signaling through exchange protein activated by cAMP and, to a lesser degree, protein kinase A. In addition to effects on GrB, VIP also down-regulates Fas ligand (FasL) and perforin (Pfr) expression. The extrinsic Fas/FasL pathway and the intrinsic GrB-dependent pathway act independently in inducing AICD. The mechanisms by which GrB induces cell death in Th1/Th2 effectors include both fratricide and suicide. Fratricide killing, prevalent in wild-type cells, is calcium and Pfr dependent, whereas the cell death of Pfr-deficient Th cells involves Fas and GrB but is calcium independent. This study identifies GrB as a new significant player in Th1/Th2 AICD and characterizes two mechanisms for the protective effect of VIP on Th2 survival, i.e., the down-regulation of GrB and FasL expression. PMID- 16365401 TI - alpha-Lipoic acid inhibits inflammatory bone resorption by suppressing prostaglandin E2 synthesis. AB - alpha-Lipoic acid (LA) has been intensely investigated as a therapeutic agent for several pathological conditions, including diabetic polyneuropathy. In the present study, we examined the effects of LA on osteoclastic bone loss associated with inflammation. LA significantly inhibited IL-1-induced osteoclast formation in cocultures of mouse osteoblasts and bone marrow cells, but LA had only a marginal effect on osteoclastogenesis from bone marrow macrophages induced by receptor activator of NF-kappaB ligand (RANKL). LA inhibited both the sustained up-regulation of RANKL expression and the production of PGE2 induced by IL-1 in osteoblasts. In addition, treatment with either prostaglandin E2 (PGE2) or RANKL rescued IL-1-induced osteoclast formation inhibited by LA or NS398, a specific cyclooxygenase-2 (COX-2) inhibitor, in cocultures. LA blocked IL-1-induced PGE2 production even in the presence of arachidonic acid, without affecting the expression of COX-2 and membrane-bound PGE2 synthase. Dihydrolipoic acid (the reduced form of LA), but not LA, attenuated recombinant COX-2 activity in vitro. LA also inhibited osteoclast formation and bone loss induced by IL-1 and LPS in mice. Our results suggest that the reduced form of LA inhibits COX-2 activity, PGE2 production, and sustained RANKL expression, thereby inhibiting osteoclast formation and bone loss in inflammatory conditions. PMID- 16365402 TI - B7+ iris pigment epithelium induce CD8+ T regulatory cells; both suppress CTLA-4+ T cells. AB - Ocular pigment epithelia contribute to immune privilege by suppressing T cell activation and converting T cells into regulatory T regulatory cells (Tregs) that inhibit bystander T cell activation. Iris pigment epithelium (IPE) does so through direct cell-cell contact with naive T cells, and this suppressive contact is via interactions between B7 expressed constitutively on IPE cells and CTLA-4 expressed on a subpopulation of CD8+ T cells. We have now examined whether TGFbeta is required in this process. We report that IPE produces both soluble and membrane-bound active TGFbeta, but that only the latter is actually delivered to CD8+ T cells. In turn, these T cells become IPE Tregs by up-regulating their own expression of B7-1/B7-2 and soluble and membrane-bound TGFbeta. IPE Tregs through their expression of B7 are able to engage CTLA-4+ bystander T cells, and thus precisely, target delivery of membrane-bound TGFbeta. We propose that this mechanism of suppression via TGFbeta ensures that soluble active TGFbeta is not released into the ocular microenvironment where it can have unregulated and deleterious effects, including elevation of intraocular pressure and development of glaucoma. PMID- 16365403 TI - Allograft rejection requires STAT5a/b-regulated antiapoptotic activity in T cells but not B cells. AB - STATs play key roles in immune function. We examined the role of STAT5a/b in allograft rejection. STAT5a/b-deficient mice showed a 4-fold increased survival time of heart allografts (p < 0.01). Unlike wild type, purified STAT5a/b-/- T cells transferred to Rag1-/- recipients failed to mediate heart allograft rejection until supplemented with STAT5a/b-/- B cells. In vitro, STAT5a/b-/- T cells did not proliferate in response to Con A or alloantigens but entered apoptosis within 48 h (95%). Activated STAT5a/b-/- T cells showed increased expression of proapoptotic (caspases, DNA repair genes, TNF/TNFR-associated factor family genes) and decreased antiapoptotic mRNAs in microarrays, while Western blots confirmed reduced antiapoptotic Bcl-2 and elevated proapoptotic Bax protein expression. Interestingly, at 24 h postactivation, STAT5a/b+/+ and STAT5a/b-/- T cells produced similar levels of IL-2, IL-4, IL-10, and IFN-gamma mRNA; ELISPOT assay showed an equivalent number of IL-4- and IFN-gamma-producing T cells in both STAT5a/b+/+ and STAT5a/b-/- splenic populations. Sera from STAT5a/b+/+ and STAT5a/b-/- rejectors had donor-specific IgM, IgG1, IgG2a, and IgG2b Ab, while STAT5a/b deficiency had no impact on B cell survival or proliferation in response to LPS. Compared with allografts from STAT5a/b+/+ recipients, heart allografts from STAT5a/b-/- recipients had markedly reduced infiltration by CD4 and CD8 T cells but increased infiltration by B cells and dense endothelial deposition of C4d, a marker of humoral rejection. Thus, activated STAT5a/b-/- T cells produce cytokines prior to entering apoptosis, thereby promoting differentiation of B cells yielding donor-specific IgM and IgG Ab that mediate allograft rejection. PMID- 16365404 TI - Helminth-modified pulmonary immune response protects mice from allergen-induced airway hyperresponsiveness. AB - It has been shown that the presence of certain helminth infections in humans, including schistosomes, may reduce the propensity to develop allergies in infected populations. Using a mouse model of schistosome worm vs worm + egg infection, our objective was to dissect the mechanisms underlying the inverse relationship between helminth infections and allergies. We have demonstrated that conventional Schistosoma mansoni egg-laying male and female worm infection of mice exacerbates airway hyperresponsiveness. In contrast, mice infected with only schistosome male worms, precluding egg production, were protected from OVA induced airway hyperresponsiveness. Worm-infected mice developed a novel modified type 2 cytokine response in the lungs, with elevated allergen-specific IL-4 and IL-13 but reduced IL-5, and increased IL-10. Although schistosome worm-only infection is a laboratory model, these data illustrate the complexity of schistosome modulation of host immunity by the worm vs egg stages of this helminth, with the potential of infections to aggravate or suppress allergic pulmonary inflammation. Thus, infection of mice with a human parasitic worm can result in reduced airway inflammation in response to a model allergen. PMID- 16365405 TI - Differential gene expression in endometrium, endometrial lymphocytes, and trophoblasts during successful and abortive embryo implantation. AB - Prenatal mortality reaching 30% occurs during the first weeks of gestation in commercial swine. Mechanisms for this are unknown although poor uterine blood supply has been postulated. In other species, vascular endothelial growth factor, hypoxia-inducible factor 1-alpha, and IFN-gamma regulate gestational endometrial angiogenesis. Vascular endothelial growth factor and hypoxia-inducible factor 1 alpha are also important for placental angiogenesis while trophoblastic expression of Fas ligand is thought to protect conceptuses against immune mediated pregnancy loss. In this study, we document dynamic, peri-implantation differences in transcription of genes for angiogenesis, cytokine production, and apoptosis regulation in the endometrium, and laser capture microdissected endometrial lymphocytes and trophoblasts associated with healthy or viable but arresting porcine fetuses. In healthy implantation sites, endometrial gene expression levels differed between anatomic subregions and endometrial lymphocytes showed much greater transcription of angiogenic genes than trophoblasts. In arresting fetal sites, uterine lymphocytes had no angiogenic gene transcription and showed rapid elevation in transcription of proinflammatory cytokines Fas and Fas ligand while trophoblasts showed elevated transcription of IFN-gamma and Fas. This model of experimentally accessible spontaneous fetal loss, involving blocked maternal angiogenesis, should prove valuable for further investigations of peri-implantation failure of normally conceived and surgically transferred embryos in many species, including the human. PMID- 16365407 TI - Reduced ability of neonatal and early-life bone marrow stromal cells to support plasmablast survival. AB - In human infants (<1 year), circulating IgG Abs elicited in response to most T dependent Ags rapidly decline and return to baseline within a few months after immunization for yet-unknown reasons. In mice immunized between 1 and 4 wk of age, a limited establishment of the bone marrow (BM) pool of long-lived plasma cells is observed. In this study, we show that tetanus toxoid (TT)-specific plasmablasts generated in the spleen are efficiently attracted in vitro and in vivo toward early-life BM stromal cells, which express adult levels of CXCL12. Similarly, adoptively transferred TT plasmablasts efficiently reach the BM compartment of 2-wk-old and adult mice. In contrast, TT plasmablasts fail to persist in the early-life BM compartment, as indicated by the persistence of a significantly lower number of TT plasmablasts in the early-life compartment than in the adult BM compartment 48 h after transfer. This limited persistence is associated with an increased rate of in vivo apoptosis of TT-specific plasmablasts that have reached the early-life BM and with a significantly lower survival rate of TT-specific plasmablasts cocultured on early-life BM stromal cells compared with adult BM stromal cells. Thus, early-life BM stromal cells fail to provide the molecular signals that support plasmablast survival and differentiation into surviving plasma cells. PMID- 16365406 TI - The TLR-7 agonist, imiquimod, enhances dendritic cell survival and promotes tumor antigen-specific T cell priming: relation to central nervous system antitumor immunity. AB - Immunotherapy represents an appealing option to specifically target CNS tumors using the immune system. In this report, we tested whether adjunctive treatment with the TLR-7 agonist imiquimod could augment antitumor immune responsiveness in CNS tumor-bearing mice treated with human gp100 + tyrosine-related protein-2 melanoma-associated Ag peptide-pulsed dendritic cell (DC) vaccination. Treatment of mice with 5% imiquimod resulted in synergistic reduction in CNS tumor growth compared with melanoma-associated Ag-pulsed DC vaccination alone. Continuous imiquimod administration in CNS tumor-bearing mice, however, was associated with the appearance of robust innate immune cell infiltration and hemorrhage into the brain and the tumor. To understand the immunological mechanisms by which imiquimod augmented antitumor immunity, we tested whether imiquimod treatment enhanced DC function or the priming of tumor-specific CD8+ T cells in vivo. With bioluminescent, in vivo imaging, we determined that imiquimod dramatically enhanced both the persistence and trafficking of DCs into the draining lymph nodes after vaccination. We additionally demonstrated that imiquimod administration significantly increased the accumulation of tumor-specific CD8+ T cells in the spleen and draining lymph nodes after DC vaccination. The results suggest that imiquimod positively influences DC trafficking and the priming of tumor-specific CD8+ T cells. However, inflammatory responses induced in the brain by TLR signaling must also take into account the local microenvironment in the context of antitumor immunity to induce clinical benefit. Nevertheless, immunotherapeutic targeting of malignant CNS tumors may be enhanced by the administration of the innate immune response modifier imiquimod. PMID- 16365408 TI - The glycosaminoglycan-binding domain of decoy receptor 3 is essential for induction of monocyte adhesion. AB - Decoy receptor 3 (DcR3), a soluble receptor for Fas ligand, LIGHT (homologous to lymphotoxins shows inducible expression and competes with HSV glycoprotein D for herpes virus entry mediator, a receptor expressed by T lymphocytes), and TNF-like molecule 1A, is highly expressed in cancer cells and in tissues affected by autoimmune disease. DcR3.Fc has been shown to stimulate cell adhesion and to modulate cell activation and differentiation by triggering multiple signaling cascades that are independent of its three known ligands. In this study we found that DcR3.Fc-induced cell adhesion was inhibited by heparin and heparan sulfate, and that DcR3.Fc was unable to bind Chinese hamster ovary K1 mutants defective in glycosaminoglycan (GAG) synthesis. Furthermore, the negatively charged, sulfated GAGs of cell surface proteoglycans, but not their core proteins, were identified as the binding sites for DcR3.Fc. A potential GAG-binding site was found in the C terminal region of DcR3, and the mutation of three basic residues, i.e., K256, R258, and R259, to alanines abolished its ability to trigger cell adhesion. Moreover, a fusion protein comprising the GAG-binding region of DcR3 with an Fc fragment (DcR3_HBD.Fc) has the same effect as DcR3.Fc in activating protein kinase C and inducing cell adhesion. Compared with wild-type THP-1 cells, cell adhesion induced by DcR3.Fc was significantly reduced in both CD44v3 and syndecan 2 knockdown THP-1 cells. Therefore, we propose a model in which DcR3.Fc may bind to and cross-link proteoglycans to induce monocyte adhesion. PMID- 16365409 TI - Gap junction-mediated intercellular communication between dendritic cells (DCs) is required for effective activation of DCs. AB - Gap junctions, formed by members of the connexin (Cx) family, are intercellular channels allowing direct exchange of signaling molecules. Gap junction-mediated intercellular communication (GJIC) is a widespread mechanism for homeostasis in organs. GJIC in the immune system is not yet fully understood. Although dendritic cells (DC) reportedly form cell-to-cell contact between DCs in nonlymphoid and lymphoid organs, GJIC between DCs remains unknown. In this study we examined whether DCs form GJIC. XS52 and bone marrow-derived DCs (BMDCs) were tested for GJIC by counting intercellular transfer of Lucifer Yellow microinjected into a cell. Either DC became effectively dye-coupled when activated with LPS plus IFN gamma or TNF-alpha plus IFN-gamma. LPS- plus IFN-gamma-induced dye-coupling was mediated by DC-derived TNF-alpha. In addition, CpG plus IFN-gamma induced dye coupling in BMDCs, which was also mediated by DC-derived TNF-alpha. LPS- plus IFN gamma-induced activation of DCs (assessed by CD40 expression) was observed when there was cell-to-cell contact and was significantly blocked by heptanol, a gap junction blocker. These results indicate that cell-to-cell contact and GJIC are required for effective DC activation. In addition, heptanol significantly inhibited the LPS- plus IFN-gamma-induced up-regulation of the other costimulatory (i.e., CD80 and CD86) and MHC class II molecules expressed by BMDCs, and it significantly reduced their allostimulatory capacity. Among Cx members, Cx43 was up-regulated in dye-coupled BMDCs, and Cx mimetic peptide, a blocker of Cx-mediated GJIC, significantly inhibited the dye-coupling and activation, suggesting the involvement of Cx43. Thus, our study provides the first evidence for GJIC between DCs, which is required for effective DC activation. PMID- 16365410 TI - Regulation of myeloid cell function through the CD200 receptor. AB - Myeloid cells play pivotal roles in chronic inflammatory diseases through their broad proinflammatory, destructive, and remodeling capacities. CD200 is widely expressed on a variety of cell types, while the recently identified CD200R is expressed on myeloid cells and T cells. CD200 deletion in vivo results in myeloid cell dysregulation and enhanced susceptibility to autoimmune inflammation, suggesting that the CD200-CD200R interaction is involved in immune suppression. We demonstrate in this study that CD200R agonists suppress mouse and human myeloid cell function in vitro, and also define a dose relationship between receptor expression and cellular inhibition. IFN-gamma- and IL-17-stimulated cytokine secretion from mouse peritoneal macrophages was inhibited by CD200R engagement. Inhibitory effects were not universal, as LPS-stimulated responses were unaffected. Inhibition of U937 cell cytokine production correlated with CD200R expression levels, and inhibition was only observed in low CD200R expressing cells, if the CD200R agonists were further cross-linked. Tetanus toxoid-induced human PBMC IL-5 and IL-13 secretion was inhibited by CD200R agonists. This inhibition was dependent upon cross-linking the CD200R on monocytes, but not on cross-linking the CD200R on CD4+ T cells. In all, we provide direct evidence that the CD200-CD200R interaction controls monocyte/macrophage function in both murine and human systems, further supporting the potential clinical application of CD200R agonists for the treatment of chronic inflammatory diseases. PMID- 16365411 TI - The CD8+ T cell population elicited by recombinant adenovirus displays a novel partially exhausted phenotype associated with prolonged antigen presentation that nonetheless provides long-term immunity. AB - We have previously reported that the CD8+ T cell response elicited by recombinant adenovirus vaccination displayed a delayed contraction in the spleen. In our current study, we demonstrate that this unusual kinetic is a general phenomenon observed in multiple tissues. Phenotypic analysis of transgene-specific CD8+ T cells present 30 days postimmunization with recombinant adenovirus revealed a population with evidence of partial exhaustion, suggesting that the cells had been chronically exposed to Ag. Although Ag expression could no longer be detected 3 wk after immunization, examination of Ag presentation within the draining lymph nodes demonstrated that APCs were loaded with Ag peptide for at least 40 days postimmunization, suggesting that Ag remains available to the system for a prolonged period, although the exact source of this Ag remains to be determined. At 60 days postimmunization, the CD8+ T cell population continued to exhibit a phenotype consistent with partially exhausted effector memory cells. Nonetheless, these CD8+ T cells conferred sterilizing immunity against virus challenge 7-12 wk postimmunization, suggesting that robust protective immunity can be provided by CD8+ T cells with an exhausted phenotype. These data demonstrate that prolonged exposure to Ag may not necessarily impair protective immunity and prompt a re-evaluation of the impact of persistent exposure to Ag on T cell function. PMID- 16365412 TI - Expression of CD161 (NKR-P1A) defines subsets of human CD4 and CD8 T cells with different functional activities. AB - A subset of T cells in human peripheral blood expresses CD161 (NKR-P1A) receptors that are primarily associated with NK cells. In the current study we isolated blood T cell subsets according to the expression of CD161 and examined their contents of naive, central memory, and effector memory cells and their capacities for proliferation, cytokine secretion, and natural cytolysis. We found that CD4+CD161- and CD8+CD161- subsets contained predominantly naive T cells that secreted high levels of IL-2 after in vitro stimulation, and CD4+CD161int and CD8+CD161int subsets contained predominantly effector and central memory T cells that secreted high levels of IFN-gamma and TNF-alpha. All of these subsets showed vigorous proliferation after stimulation in vitro, but none had NK lytic activity. Unexpectedly, the CD8+CD161+ cells contained an anergic CD8alpha+CD8betalow/-CD161high T cell subset that failed to proliferate, secrete cytokines, or mediate NK lytic activity. PMID- 16365413 TI - Induction of antitumor immunity by CTL epitopes genetically linked to membrane anchored beta2-microglobulin. AB - Level and persistence of antigenic peptides presented by APCs on MHC class I (MHC I) molecules influence the magnitude and quality of the ensuing CTL response. We recently demonstrated the unique immunological properties conferred on APCs by expressing beta2-microglobulin (beta2m) as an integral membrane protein. In this study, we explored membrane-anchored beta2m as a platform for cancer vaccines using as a model MO5, an OVA-expressing mouse B16 melanoma. We expressed in mouse RMA-S cells two H-2Kb binding peptides from MO5, OVA257-264, and TRP-2181-188, each genetically fused with the N terminus of membranal beta2m via a short linker. Specific Ab staining and T cell hybridoma activation confirmed that OVA257-264 was properly situated in the MHC-I binding groove. In vivo, transfectants expressing both peptides elicited stronger CTLs and conferred better protection against MO5 than peptide-saturated RMA-S cells. Cells expressing OVA257-264/beta2m were significantly superior to OVA257-264-charged cells in their ability to inhibit the growth of pre-established MO5 tumors. Our results highlight the immunotherapeutic potential of membranal beta2m as a universal scaffold for optimizing Ag presentation by MHC-I molecules. PMID- 16365414 TI - Enhanced regulatory T cell activity is an element of the host response to injury. AB - CD4+CD25+ regulatory T cells (Tregs) play a critical role in suppressing the development of autoimmune disease, in controlling potentially harmful inflammatory responses, and in maintaining immune homeostasis. Because severe injury triggers both excessive inflammation and suppressed adaptive immunity, we wished to test whether injury could influence Treg activity. Using a mouse burn injury model, we demonstrate that injury significantly enhances Treg function. This increase in Treg activity is apparent at 7 days after injury and is restricted to lymph node CD4+CD25+ T cells draining the injury site. Moreover, we show that this injury-induced increase in Treg activity is cell-contact dependent and is mediated in part by increased cell surface TGF-beta1 expression. To test the in vivo significance of these findings, mice were depleted of CD4+CD25+ T cells before sham or burn injury and then were immunized to follow the development of T cell-dependent Ag-specific immune reactivity. We observed that injured mice, which normally demonstrate suppressed Th1-type immunity, showed normal Th1 responses when depleted of CD4+CD25+ T cells. Taken together, these observations suggest that injury can induce or amplify CD4+CD25+ Treg function and that CD4+CD25+ T cells contribute to the development of postinjury immune suppression. PMID- 16365415 TI - IL-27 limits IL-2 production during Th1 differentiation. AB - Although the ability of IL-27 to promote T cell responses is well documented, the anti-inflammatory properties of this cytokine remain poorly understood. The current work demonstrates that during infection with Toxoplasma gondii, IL-27R deficient mice generate aberrant IL-2 responses that are associated with the development of a lethal inflammatory disease. Because in vivo depletion of IL-2 prolongs the survival of infected IL-27R-/- mice, these data suggest that IL-27 curbs the development of immunopathology by limiting parasite-induced IL-2 production. Consistent with this hypothesis, IL-27R-/- CD4+ T cells produce more IL-2 than wild-type counterparts during in vitro differentiation, and when rIL-27 is introduced, it can suppress the expression of IL-2 mRNA and protein by the latter group. Additionally, these studies reveal that, like IL-27, IL-12 can inhibit IL-2 production, and although each employs distinct mechanisms, they can synergize to enhance the effect. In contrast, this property is not shared by closely related cytokines IL-6 and IL-23. Thus, while traditionally viewed as proinflammatory agents, the present findings establish that IL-27 and IL-12 cooperate to limit the availability of IL-2, a potent T cell growth and survival factor. Moreover, because the current studies demonstrate that both can induce expression of suppressor of cytokine signaling 3, a protein that tempers cytokine receptor signaling, they also suggest that IL-27 and IL-12 share additionally inhibitory properties. PMID- 16365416 TI - Virus or TLR agonists induce TRAIL-mediated cytotoxic activity of plasmacytoid dendritic cells. AB - Among dendritic cells, plasmacytoid dendritic cells (PDC) represent a functionally distinct lineage. Regarding innate immunity, PDC secrete large amounts of type I IFN upon viral exposure or stimulation by microbial products such as unmethylated CpG-motif containing oligo-DNA due to their selective expression of TLR7 and TLR9. We asked whether they could acquire cytotoxic functions during the early phases of infection or after activation with TLR7 or TLR9 agonists. In the present study, we describe a human PDC cell line called GEN2.2, derived from leukemic PDC, that shares most of the phenotypic and functional features of normal PDC. We show that after contact with the influenza virus, GEN2.2, as well as normal PDC, acquires TRAIL and killer activity against TRAIL-sensitive target cells. Moreover, we show that activation of GEN2.2 cells by CpG-motif containing oligo-DNA or R848 also induces TRAIL and endows them with the ability to kill melanoma cells. Therefore, PDC may represent a major component of innate immunity that could participate to the clearance of infected cells and tumor cells. This phenomenon could be relevant for the efficacy of TLR7 or TLR9 agonists in the therapy of infectious disease and cancer. PMID- 16365417 TI - The novel cytokine p43 induces IL-12 production in macrophages via NF-kappaB activation, leading to enhanced IFN-gamma production in CD4+ T cells. AB - Recently, we determined that p43, an auxiliary factor of mammalian multiaminoacyl tRNA synthetases, is secreted, and functions as a novel pleiotropic cytokine. In this study, we have attempted to characterize the effects of p43 on the generation of IL-12 in mouse macrophages. p43 was determined to induce significant IL-12 production from mouse macrophages in a dose-dependent manner. The stimulatory effect of p43 on the activation of IL-12p40 promoter was mapped to a region harboring an NF-kappaB binding site. The nuclear extracts from the p43-stimulated macrophages exhibited profound NF-kappaB DNA-binding activity, as determined by the EMSA. In addition, the p43-stimulated IL-12 induction and NF kappaB DNA-binding activity were significantly suppressed by caffeic acid phenethyl ester and BAY11-7082, both inhibitors of NF-kappaB activation, indicating that p43 induced the production of IL-12 in macrophages mainly via the activation of NF-kappaB. Importantly, p43 increased the level of IFN-gamma production in the Ag-primed lymph node cells, but had no effect on IL-4 levels. The addition of a neutralizing anti-IL-12p40 mAb to the cell cultures resulted in a decrease of the production of p43-enhanced IFN-gamma by the keyhole limpet hemocyanin-primed lymph node cells. Furthermore, coincubation with p43-pretreated macrophages enhanced the production of IFN-gamma by the keyhole limpet hemocyanin primed CD4+ T cells, thereby indicating that p43 may enhance IFN-gamma expression in CD4+ T cells via the induction of IL-12 production in macrophages. These results indicate that p43 may play an essential role in the development of the Th1 immune responses associated with cancer immunotherapy and protective immunity against intracellular pathogens. PMID- 16365418 TI - Cellular and molecular mechanisms of the selective regulation of IL-12 production by 12/15-lipoxygenase. AB - IL-12 drives type I immune responses and can mediate chronic inflammation that leads to host defense as well as disease. Recently, we discovered a novel role for 12/15-lipoxygenase (12/15-LO) in mediating IL-12p40 expression in atherosclerotic plaque and in isolated macrophages. We now demonstrate that 12/15 LO regulates IL-12 family cytokine production in a cell-type and stimulus restricted fashion. LPS-stimulated elicited peritoneal macrophages derived from 12/15-LO-deficient (Alox15) mice produced reduced IL-12 and IL-23 levels, but comparable amounts of several other inflammatory mediators tested. Furthermore, LPS stimulation triggered an increase in wild-type macrophage 12/15-LO activity, whereas pharmacological inhibition of 12/15-LO activity suppressed LPS-induced IL 12 production in wild-type macrophages. 12/15-LO-deficient macrophages also produced reduced levels of IL-12 in response to TLR2 stimulation, but not in response to CpG (TLR9) or CD40/CD40L-mediated activation. In contrast to our previous finding of reduced IL-12 production in the setting of atherosclerosis, we found that comparable IL-12 levels were produced in Alox15 and wild-type mice during an acute response to LPS in vivo. This paradox may be explained by normal production of IL-12 by 12/15-LO-deficient neutrophils and dendritic cells, which are major sources of IL-12 during acute inflammation. Finally, we detected selectively decreased association of the transcription factors IFN consensus sequence binding protein and NF-kappaB with the IL-12p40 promoter in 12/15-LO deficient macrophages. Taken together, these findings reveal a highly selective pathway to IL-12 production that may prove a useful target in chronic inflammation while sparing the acute response to infection. PMID- 16365419 TI - PU.1 regulates cathepsin S expression in professional APCs. AB - Cathepsin S (CTSS) is a cysteine protease that is constitutively expressed in APCs and mediates processing of MHC class II-associated invariant chain. CTSS and the Ets family transcription factor PU.1 are highly expressed in cells of both myeloid (macrophages and dendritic cells) and lymphoid (B lymphocytes) lineages. Therefore, we hypothesized that PU.1 participates in the transcriptional regulation of CTSS in these cells. In A549 cells (a human epithelial cell line that does not express either CTSS or PU.1), the expression of PU.1 enhances CTSS promoter activity approximately 5- to 10-fold. In RAW cells (a murine macrophage like cell line that constitutively expresses both CTSS and PU.1), the expression of a dominant-negative PU.1 protein and a short-interfering RNA PU.1 construct attenuates basal CTSS promoter activity, mRNA levels, and protein expression. EMSAs show binding of PU.1 to oligonucleotides derived from the CTSS promoter at two different Ets consensus binding elements. Mutation of these sites decreases the baseline CTSS activity in RAW cells that constitutively express PU.1. Chromatin immunoprecipitation experiments show binding of PU.1 with the CTSS promoter in this same region. Finally, the expression of PU.1, in concert with several members of the IFN regulatory factor family, enhances CTSS promoter activity beyond that achieved by PU.1 alone. These data indicate that PU.1 participates in the regulation of CTSS transcription in APCs. Thus, manipulation of PU.1 expression may directly alter the endosomal proteolytic environment in these cells. PMID- 16365420 TI - Inflammation induces myeloid-derived suppressor cells that facilitate tumor progression. AB - Epidemiological and experimental observations support the hypothesis that chronic inflammation contributes to cancer development and progression; however, the mechanisms underlying the relationship between inflammation and cancer are poorly understood. To study these mechanisms, we have transfected the mouse 4T1 mammary carcinoma with the proinflammatory cytokine IL-1beta to produce a chronic inflammatory microenvironment at the tumor site. Mice with 4T1/IL-1beta tumors have a decreased survival time and elevated levels of immature splenic Gr1+CD11b+ myeloid-derived cells. These myeloid suppressor cells (MSC) are present in many patients with cancer and inhibit the activation of CD4+ and CD8+ T lymphocytes. 4T1/IL-1beta-induced MSC do not express the IL-1R, suggesting that the cytokine does not directly activate MSC. Neither T or B cells nor NKT cells are involved in the IL-1beta-induced increase of MSC because RAG2-/- mice and nude mice with 4T1/IL-1beta tumors also have elevated MSC levels. MSC levels remain elevated in mice inoculated with 4T1/IL-1beta even after the primary tumor is surgically removed, indicating that the IL-1beta effect is long lived. Collectively, these findings suggest that inflammation promotes malignancy via proinflammatory cytokines, such as IL-1beta, which enhance immune suppression through the induction of MSC, thereby counteracting immune surveillance and allowing the outgrowth and proliferation of malignant cells. PMID- 16365421 TI - Ly9 (CD229)-deficient mice exhibit T cell defects yet do not share several phenotypic characteristics associated with SLAM- and SAP-deficient mice. AB - Signaling lymphocyte activation molecule (SLAM) family receptors are critically involved in modulating innate and adaptive immune responses. Several SLAM family receptors have been shown to interact with the adaptor molecule SAP; however, subsequent intracellular signaling is poorly defined. Notably, mutations in SLAM associated protein (SAP) lead to X-linked lymphoproliferative disease, a rare but fatal immunodeficiency. Although the SLAM family member Ly9 (CD229) is known to interact with SAP, the functions of this receptor have remained elusive. Therefore, we have generated Ly9-/- mice and compared their phenotype with that of SLAM-/- and SAP-/- mice. We report that Ly9-/- T cells exhibit a mild Th2 defect associated with reduced IL-4 production after stimulation with anti-TCR and anti-CD28 in vitro. This defect is similar in magnitude to the previously reported Th2 defect in SLAM-/- mice but is more subtle than that observed in SAP /- mice. In contrast to SLAM-/- and SAP-/- mice, T cells from Ly9-/- mice proliferate poorly and produce little IL-2 after suboptimal stimulation with anti CD3 in vitro. We have also found that Ly9-/- macrophages exhibit no defects in cytokine production or bacterial killing as was observed in SLAM-/- macrophages. Additionally, Ly9-/- mice differ from SAP-/- mice in that they foster normal development of NKT cells and mount appropriate T and B cell responses to lymphocytic choriomeningitis virus. We have identified significant phenotypic differences between Ly-9-/- mice as compared with both SLAM-/- and SAP-/- mice. Although Ly9, SLAM, and SAP play a common role in promoting Th2 polarization, Ly 9 is uniquely involved in enhancing T cell activation. PMID- 16365422 TI - Molecular, cellular, and antigen requirements for development of age-associated T cell clonal expansions in vivo. AB - T cell aging manifests itself both at the cellular (cell-autonomous defects in signaling) and at the population (age-related dysregulation of T cell homeostasis) levels. A prominent contributor to the latter is the appearance of T cell clonal expansions (TCE), with a potential to impair immune defense. In this study, we investigated molecular, cellular, and Ag requirements for TCE development. Of the mutant mice tested, old animals lacking MHC class I exhibited 7-fold fewer TCE than controls, with a 7-fold reduction in TCE. By contrast, animals lacking only one of the MHC class I molecules (Kb or Db), or IL-7R, or devoid of T cell renewal via adult thymectomy, all exhibited significant increases in TCE incidence. This increase directly correlated to lymphopenia, increased CD8 T cell turnover and an accumulation of memory-phenotype T cells. These data suggested that homeostatic cell division in the CD8 compartment enhances the formation of TCE. Repeated immunization with peptide/adjuvant did not result in an increase in Ag-specific TCE; however, adjuvant alone increased TCE incidence. In these experiments, therefore, nonspecific and/or homeostatic proliferation was more efficient in generating TCE in mice than repeated Ag driven stimulation, suggesting that many, if not most, TCE in specific pathogen free laboratory mice may be Ag-independent. PMID- 16365423 TI - Synergistic activation of macrophages via CD40 and TLR9 results in T cell independent antitumor effects. AB - We have previously shown that macrophages (Mphi) can be activated by CD40 ligation to become cytotoxic against tumor cells in vitro. Here we show that treatment of mice with agonistic anti-CD40 mAb (anti-CD40) induced up-regulation of intracellular TLR9 in Mphi and primed them to respond to CpG-containing oligodeoxynucleotides (CpG), resulting in synergistic activation. The synergy between anti-CD40 and CpG was evidenced by increased production of IFN-gamma, IL 12, TNF-alpha, and NO by Mphi, as well as by augmented apoptogenic effects of Mphi against tumor cells in vitro. The activation of cytotoxic Mphi after anti CD40 plus CpG treatment was dependent on IFN-gamma but not TNF-alpha or NO, and did not require T cells and NK cells. Anti-CD40 and CpG also synergized in vivo in retardation of tumor growth in both immunocompetent and immunodeficient mice. Inactivation of Mphi in SCID/beige mice by silica treatment abrogated the antitumor effect. Taken together, our results show that Mphi can be activated via CD40/TLR9 ligation to kill tumor cells in vitro and inhibit tumor growth in vivo even in immunocompromised tumor-bearing hosts, indicating that this Mphi-based immunotherapeutic strategy may be appropriate for clinical testing. PMID- 16365424 TI - Immunodomination in the evolution of dominant epitope-specific CD8+ T lymphocyte responses in simian immunodeficiency virus-infected rhesus monkeys. AB - Because the control of HIV-1 replication is largely dependent on CD8+ T lymphocyte responses specific for immunodominant viral epitopes, vaccine strategies that increase the breadth of dominant epitope-specific responses should contribute to containing HIV-1 spread. Developing strategies to elicit such broad immune responses will require an understanding of the mechanisms responsible for focusing CD8+ T lymphocyte recognition on a limited number of epitopes. To explore this biology, we identified cohorts of rhesus monkeys that expressed the MHC class I molecules Mamu-A*01, Mamu-A*02, or both, and assessed the evolution of their dominant epitope-specific CD8+ T lymphocyte responses (Gag p11C- and Tat TL8-specific in the Mamu-A*01+ and Nef p199RY-specific in the Mamu A*02+ monkeys) following acute SIV infection. The Mamu-A*02+ monkeys that also expressed Mamu-A*01 exhibited a significant delay in the evolution of the CD8+ T lymphocyte responses specific for the dominant Mamu-A*02-restricted SIV epitope, Nef p199RY. This delay in kinetics was not due to differences in viral load kinetics or magnitude or in viral escape mutations, but was associated with the evolution of the Mamu-A*01-restricted CD8+ T lymphocyte responses to the highly dominant SIV epitopes Gag p11C and Tat TL8. Thus, the evolution of dominant epitope-specific CD8+ T lymphocyte responses can be suppressed by other dominant epitope-specific responses, and this immunodomination is important in determining the kinetics of dominant epitope-specific responses. PMID- 16365425 TI - Specificity of CD4+CD25+ regulatory T cell function in alloimmunity. AB - CD4+CD25+ regulatory T cells (TRegs) are critical for the acquisition of peripheral allograft tolerance. However, it is unclear whether TRegs are capable of mediating alloantigen-specific suppressive effects and, hence, contributing to the specificity of the tolerant state. In the current report we have used the ABM TCR transgenic (Tg) system, a C57BL/6-derived strain in which CD4+ T cells directly recognize the allogeneic MHC-II molecule I-A(bm12), to assess the capacity of TRegs to mediate allospecific effects. In these mice, 5-6% of Tg CD4+ T cells exhibit conventional markers of the TReg phenotype. ABM TRegs are more effective than wild-type polyclonal TRegs at suppressing effector immune responses directed against I-A(bm12) alloantigen both in vitro and in vivo. In contrast, they are incapable of suppressing responses directed against third party alloantigens unless these are expressed in the same allograft as I-A(bm12). Taken together, our results indicate that in transplantation, TReg function is dependent on TCR stimulation, providing definitive evidence for their specificity in the regulation of alloimmune responses. PMID- 16365426 TI - SRC-like adaptor protein regulates B cell development and function. AB - The avidity of BCRs and TCRs influences signal strength during processes of lymphocyte development. Avidity is determined by both the intrinsic affinity for Ag and surface levels of the Ag receptor. The Src-like adaptor protein (SLAP) is a regulator of TCR levels on thymocytes, and its deficiency alters thymocyte development. We hypothesized that SLAP, which is expressed in B cells, also is important in regulating BCR levels, signal strength, and B cell development. To test this hypothesis, we analyzed the B cell compartment in SLAP-deficient mice. We found increased splenic B cell numbers and decreased surface IgM levels on mature, splenic B cells deficient in SLAP. Immature bone marrow and splenic B cells from BCR-transgenic, SLAP-deficient mice were found to express higher surface levels of IgM. In contrast, mature splenic B cells from BCR-transgenic mice expressed decreased levels of surface BCR associated with decreased calcium flux and activation-induced markers, compared with controls. These data suggest that SLAP regulates BCR levels and signal strength during lymphocyte development. PMID- 16365427 TI - An engineered human IgG1 antibody with longer serum half-life. AB - The serum half-life of IgG Abs is regulated by the neonatal Fc receptor (FcRn). By binding to FcRn in endosomes, IgG Abs are salvaged from lysosomal degradation and recycled to the circulation. Several studies have demonstrated a correlation between the binding affinity of IgG Abs to FcRn and their serum half-lives in mice, including engineered Ab fragments with longer serum half-lives. Our recent study extended this correlation to human IgG2 Ab variants in primates. In the current study, several human IgG1 mutants with increased binding affinity to human FcRn at pH 6.0 were generated that retained pH-dependent release. A pharmacokinetics study in rhesus monkeys of one of the IgG1 variants indicated that its serum half-life was approximately 2.5-fold longer than the wild-type Ab. Ag binding was unaffected by the Fc mutations, while several effector functions appeared to be minimally altered. These properties suggest that engineered Abs with longer serum half-lives may prove to be effective therapeutics in humans. PMID- 16365428 TI - neonatally primed lymph node, but not splenic T cells, display a Gly-Gly motif within the TCR beta-chain complementarity-determining region 3 that controls affinity and may affect lymphoid organ retention. AB - Ig-proteolipid protein 1 (Ig-PLP1) is an Ig chimera expressing the encephalitogenic PLP1 peptide corresponding to amino acid residues 139-151 of PLP. Newborn mice given Ig-PLP1 in saline on the day of birth and challenged 7 wk later with PLP1 peptide in CFA develop an organ-specific neonatal immunity that confers resistance against experimental allergic encephalomyelitis. The T cell responses in these animals are comprised of Th2 cells in the lymph node and anergic Th1 lymphocytes in the spleen. Intriguingly, the anergic splenic T cells, although nonproliferative and unable to produce IFN-gamma or IL-4, secrete significant amounts of IL-2. Studies were performed to determine whether the two populations display any structural differences in the TCR H chain variable region that could contribute to the differential affinity and retention in different organs. Responsive Th2 lymph node T cells and anergic splenic lymphocytes were immortalized, and the structures of their TCR Vbeta were determined. The results show that Vbeta and Jbeta usage was random, but the CDR3 regions of the lymph node cells had a conserved Gly-Gly motif. Analysis of TCR affinity/avidity correlated the Gly-Gly motif with lower affinity and retention of the Th2 cells in the lymph node. Also, it is suggested that a higher TCR affinity may be a contributing factor for the development of the neonatal Th1 response in the spleen. PMID- 16365429 TI - Uracil DNA glycosylase disruption blocks Ig gene conversion and induces transition mutations. AB - Ig gene conversion is most likely initiated by activation-induced cytidine deaminase-mediated cytosine deamination. If the resulting uracils need to be further processed by uracil DNA glycosylase (UNG), UNG inactivation should block gene conversion and induce transition mutations. In this study, we report that this is indeed the phenotype in the B cell line DT40. Ig gene conversion is almost completely extinguished in the UNG-deficient mutant and large numbers of transition mutations at C/G bases accumulate within the rearranged Ig L chain gene (IgL). The mutation rate of UNG-deficient cells is about seven times higher than that of pseudo V gene-deleted (psiV-) cells in which mutations arise presumably after uracil excision. In addition, UNG-deficient cells show relatively more mutations upstream and downstream of the VJ segment. This suggests that hypermutating B cells process activation-induced cytidine deaminase induced uracils with approximately one-seventh of uracils giving rise to mutations depending on their position. PMID- 16365430 TI - Nitration and inactivation of IDO by peroxynitrite. AB - IDO induction can deplete L-tryptophan in target cells, an effect partially responsible for the antimicrobial activities and antiallogeneic T cell responses of IFN-gamma in human macrophages, dendritic cells, and bone marrow cells. L tryptophan depletion and NO production are both known to have an antimicrobial effect in macrophages, and the interaction of these two mechanisms is unclear. In this study we found that IDO activity was inhibited by the peroxynitrite generator, 3-(4-morpholinyl)sydnonimine, in PMA-differentiated cytokine-induced THP-1 (acute monocytic leukemia) cells and IFN-gamma-stimulated PBMCs, whereas IDO protein expression was unaffected compared with that in untreated cells. Nitrotyrosine was detected in immunoprecipitated (IP)-IDO from PMA-differentiated cytokine-induced THP-1 cells treated with 3-(4-morpholinyl)sydnonimine, but not from untreated cells. Treatment of IP-IDO and recombinant IDO (rIDO) with peroxynitrite significantly decreased enzyme activity. Nitrotyrosine was detected in both peroxynitrite-treated IP-IDO and rIDO, but not in either untreated IP-IDO or rIDO. Peptide analysis by liquid chromatography/electrospray ionization and tandem mass spectrometry demonstrated that Tyr15, Tyr345, and Tyr353 in rIDO were nitrated by peroxynitrite. The levels of Tyr nitration and the inhibitory effect of peroxynitrite on IDO activity were significantly reduced in the Tyr15-to-Phe mutant. These results indicate that IDO is nitrated and inactivated by peroxynitrite and that nitration of Tyr15 in IDO protein is the most important factor in the inactivation of IDO. PMID- 16365431 TI - Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. AB - Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains pleckstrin and Src homology 2-like domains as well as a YXXQ motif in its C-terminal region. Our previous studies have demonstrated that STAP-2 binds to STAT3 and STAT5, and regulates their signaling pathways. In the present study, STAP-2 was found to positively regulate LPS/TLR4-mediated signals in macrophages. Disruption of STAP-2 resulted in impaired LPS/TLR4-induced cytokine production and NF-kappaB activation. Conversely, overexpression of STAP 2 enhanced these LPS/TLR4-induced biological activities. STAP-2, particularly its Src homology 2-like domain, bound to both MyD88 and IkappaB kinase (IKK) alphabeta, but not TNFR-associated factor 6 or IL-1R-associated kinase 1, and formed a functional complex composed of MyD88-STAP-2-IKK-alphabeta. These interactions augmented MyD88- and/or IKK-alphabeta-dependent signals, leading to enhancement of the NF-kappaB activity. These results demonstrate that STAP-2 may constitute an alternative LPS/TLR4 pathway for NF-kappaB activation instead of the TNFR-associated factor 6-IL-1R-associated kinase 1 pathway. PMID- 16365432 TI - Altered self-erythrocyte recognition and destruction in an inbred line of tilapia (Oreochromis aureus). AB - Carboxyfluorescein diacetate (cFDA)-stained autologous and syngeneic tilapia (Oreochromis aureus) erythrocytes are recognized by effector peripheral blood leukocytes and lysed after a short culture period of 4 h. The hemolysis level was evaluated by measuring the fluorescence of the released cFDA. The degree of lysis of stained target erythrocytes of 60 individuals revealed a trimodal distribution statistically stratified into three groups of low (LR), intermediate (IR), and high (HR) responders. Depletion of the majority of phagocytes from leukocytes lowered the lysis level of HR to that of LR. A highly significant increase of LR cytotoxicity was obtained after the addition of conditioned medium from HR but only in the presence of phagocytes. Genetic analysis of offspring from four crosses (IR x HR, IR x LR, HR x LR, and LR x LR) revealed a quantitative trait locus (QTL) segregating for the level of response linked to markers UNH207 and UNH231 on linkage group 6 of tilapia. Based on segregation analysis of 58 gynogenetic BIU-1 offspring, the distances from the centromere were estimated as 21.5, 11.5, and 9.0 cM for UNH207, UNH231, and the QTL, respectively. It is suggested that 1) self-target recognition and destruction requires both cFDA altered self-erythrocyte membrane and membrane structures normally present in autologous, syngeneic, and xenogeneic targets; 2) natural cytotoxic cells and/or macrophages are involved in erythrocyte lysis; and 3) the lysis level is codominantly inherited by a QTL segregating on tilapia linkage group 6. PMID- 16365433 TI - A signal adaptor SLAM-associated protein regulates spontaneous autoimmunity and Fas-dependent lymphoproliferation in MRL-Faslpr lupus mice. AB - Autoantibody production and lymphadenopathy are common features of systemic autoimmune disease. Targeted or spontaneous mutations in the mouse germline have generated many autoimmune models with these features. Importantly, the models have provided evidence for the gene function in prevention of autoimmunity, suggesting an indispensable role for the gene in normal immune response and homeostasis. We describe here pathological and genetic characterizations of a new mutant strain of mice, the mutation of which spontaneously occurred in the Fas deficient strain, MRL/Mp.Faslpr (MRL/lpr). MRL/lpr is known to stably exhibit systemic lupus erythematosus-like diseases. However, the mutant mice barely displayed autoimmune phenotypes, though the original defect in Fas expression was unchanged. Linkage analysis using (mutant MRL/lpr x C3H/lpr)F2 mice demonstrated a nucleotide insertion that caused loss of expression of small adaptor protein, signaling lymphocyte activation molecule (SLAM)-associated protein (SAP). SAP is known to be a downstream molecule of SLAM family receptors and to mediate the activation signal for tyrosine kinase Fyn. Recent studies have shown pleiotropic roles of SAP in T, B, and NK cell activations and NKT cell development. The present study will provide evidence for an essential role for SAP in the development of autoimmune diseases, autoantibodies, and lymphadenopathy in MRL/lpr lupus mice. PMID- 16365434 TI - Defining the origins and evolution of the chemokine/chemokine receptor system. AB - The chemokine system has a critical role in mammalian immunity, but the evolutionary history of chemokines and chemokine receptors are ill-defined. We used comparative whole genome analysis of fruit fly, sea urchin, sea squirt, pufferfish, zebrafish, frog, and chicken to identify chemokines and chemokine receptors in each species. We report 127 chemokine and 70 chemokine receptor genes in the 7 species, with zebrafish having the most chemokines, 63, and chemokine receptors, 24. Fruit fly, sea urchin, and sea squirt have no identifiable chemokines or chemokine receptors. This study represents the most comprehensive analysis of the chemokine system to date and the only complete characterization of chemokine systems outside of mouse and human. We establish a clear evolutionary model of the chemokine system and trace the origin of the chemokine system to approximately 650 million years ago, identifying critical steps in their evolution and demonstrating a more extensive chemokine system in fish than previously thought. PMID- 16365435 TI - Antimicrobial activity of native and synthetic surfactant protein B peptides. AB - Surfactant protein B (SP-B) is secreted into the airspaces with surfactant phospholipids where it reduces surface tension and prevents alveolar collapse at end expiration. SP-B is a member of the saposin-like family of proteins, several of which have antimicrobial properties. SP-B lyses negatively charged liposomes and was previously reported to inhibit the growth of Escherichia coli in vitro; however, a separate study indicated that elevated levels of SP-B in the airspaces of transgenic mice did not confer resistance to infection. The goal of this study was to assess the antimicrobial properties of native SP-B and synthetic peptides derived from the native peptide. Native SP-B aggregated and killed clinical isolates of Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and group B streptococcus by increasing membrane permeability; however, SP-B also lysed RBC, indicating that the membranolytic activity was not selective for bacteria. Both the antimicrobial and hemolytic activities of native SP-B were inhibited by surfactant phospholipids, suggesting that endogenous SP-B may not play a significant role in alveolar host defense. Synthetic peptides derived from native SP-B were effective at killing both Gram-positive and Gram-negative bacteria at low peptide concentrations (0.15-5.0 microM). The SP-B derivatives selectively lysed bacterial membranes and were more resistant to inhibition by phospholipids; furthermore, helix 1 (residues 7-22) retained significant antimicrobial activity in the presence of native surfactant. These results suggest that the role of endogenous SP-B in host defense may be limited; however, synthetic peptides derived from SP-B may be useful in the treatment of bacterial pneumonias. PMID- 16365436 TI - Internalizing antibodies to the C-type lectins, L-SIGN and DC-SIGN, inhibit viral glycoprotein binding and deliver antigen to human dendritic cells for the induction of T cell responses. AB - The C-type lectin L-SIGN is expressed on liver and lymph node endothelial cells, where it serves as a receptor for a variety of carbohydrate ligands, including ICAM-3, Ebola, and HIV. To consider targeting liver/lymph node-specific ICAM-3 grabbing nonintegrin (L-SIGN) for therapeutic purposes in autoimmunity and infectious disease, we isolated and characterized Fabs that bind strongly to L SIGN, but to a lesser degree or not at all to dendritic cell-specific ICAM grabbing nonintegrin (DC-SIGN). Six Fabs with distinct relative affinities and epitope specificities were characterized. The Fabs and those selected for conversion to IgG were tested for their ability to block ligand (HIV gp120, Ebola gp, and ICAM-3) binding. Receptor internalization upon Fab binding was evaluated on primary human liver sinusoidal endothelial cells by flow cytometry and confirmed by confocal microscopy. Although all six Fabs internalized, three Fabs that showed the most complete blocking of HIVgp120 and ICAM-3 binding to L-SIGN also internalized most efficiently. Differences among the Fab panel in the ability to efficiently block Ebola gp compared with HIVgp120 suggested distinct binding sites. As a first step to consider the potential of these Abs for Ab mediated Ag delivery, we evaluated specific peptide delivery to human dendritic cells. A durable human T cell response was induced when a tetanus toxide epitope embedded into a L-SIGN/DC-SIGN-cross-reactive Ab was targeted to dendritic cells. We believe that the isolated Abs may be useful for selective delivery of Ags to DC-SIGN- or L-SIGN-bearing APCs for the modulation of immune responses and for blocking viral infections. PMID- 16365437 TI - Interaction of mouse dendritic cells and malaria-infected erythrocytes: uptake, maturation, and antigen presentation. AB - Consistent with their seminal role in detecting infection, both mouse bone marrow derived and splenic CD11c+ dendritic cells (DCs) exhibited higher levels of uptake of Plasmodium chabaudi-parasitized RBCs (pRBCs) than of noninfected RBCs (nRBCs) as determined by our newly developed flow cytometric technique using the dye CFSE to label RBCs before coculture with DCs. To confirm that expression of CFSE by CD11c+ cells following coculture with CFSE-labeled pRBCs represents internalization of pRBC by DCs, we showed colocalization of CFSE-labeled pRBCs and PE-labeled CD11c+ DCs by confocal fluorescence microscopy. Treatment of DCs with cytochalasin D significantly inhibited the uptake of pRBCs, demonstrating that uptake is an actin-dependent phagocytic process. The uptake of pRBCs by splenic CD11c+ DCs was significantly enhanced after infection in vivo and was associated with the induction of DC maturation, IL-12 production, and stimulation of CD4+ T cell proliferation and IFN-gamma production. These results suggest that DCs selectively phagocytose pRBCs and present pRBC-derived Ags to CD4+ T cells, thereby promoting development of protective Th1-dependent immune responses to blood-stage malaria infection. PMID- 16365438 TI - The viral protein A238L inhibits TNF-alpha expression through a CBP/p300 transcriptional coactivators pathway. AB - African swine fever virus (ASFV) is able to inhibit TNF-alpha-induced gene expression through the synthesis of A238L protein. This was shown by the use of deletion mutants lacking the A238L gene from the Vero cell-adapted Ba71V ASFV strain and from the virulent isolate E70. To further analyze the molecular mechanism by which the viral gene controls TNF-alpha, we have used Jurkat cells stably transfected with the viral gene to identify the TNF-alpha regulatory elements involved in the induction of the gene after stimulation with PMA and calcium ionophore. We have thus identified the cAMP-responsive element and kappa3 sites on the TNF-alpha promoter as the responsible of the gene activation, and demonstrate that A238L inhibits TNF-alpha expression through these DNA binding sites. This inhibition was partially reverted by overexpression of the transcriptional factors NF-AT, NF-kappaB, and c-Jun. Furthermore, we present evidence that A238L inhibits the activation of TNF-alpha by modulating NF-kappaB, NF-AT, and c-Jun trans activation through a mechanism that involves CREB binding protein/p300 function, because overexpression of these transcriptional coactivators recovers TNF-alpha promoter activity. In addition, we show that A238L is a nuclear protein that binds to the cyclic AMP-responsive element/kappa3 complex, thus displacing the CREB binding protein/p300 coactivators. Taken together, these results establish a novel mechanism in the control of TNF-alpha gene expression by a viral protein that could represent an efficient strategy used by ASFV to evade the innate immune response. PMID- 16365439 TI - Effects of immunization with CCR5-based cycloimmunogen on simian/HIVSF162P3 challenge. AB - A synthetic cycloimmunogen targeting the HIV-1 coreceptor CCR5 was evaluated for its capacity to induce CCR5-specific Abs with anti-HIV-1 activity in cynomolgus macaques. The cyclic closed-chain dodecapeptide (cDDR5) mimicking the conformation-specific domain of human CCR5 was chemically prepared, in which the Gly-Glu dipeptide links the amino and carboxy termini of the decapeptidyl linear chain (Arg168 to Thr177) derived from the undecapeptidyl arch (Arg168 to Cys178) of extracellular loop-2 in CCR5. The immunization of cynomolgus macaques with the cDDR5-conjugated multiple-Ag peptide (cDDR5-MAP) induced anti-cDDR5 serum production for approximately 15 wk after the third immunization. The antisera raised against cDDR5-MAP reacted with both human and macaque CCR5s, and potently suppressed infection by the R5 HIV-1 laboratory isolate (HIV JRFL), R5 HIV-1 primary isolates (clade A:HIV 93RW004 and clade C:HIV MJ4), and a pathogenic simian/HIV (SHIV SF162P3) bulk isolate in vitro. To examine the prophylactic efficacy of anti-CCR5 serum Ab for acute HIV-1 infection, cynomolgus macaques were challenged with SHIV SF162P3. The cDDR5-MAP immunization attenuated the acute phase of SHIV SF162P3 replication. The geometric mean plasma viral load in the vaccinated macaques was 217.10 times lower than that of the control macaques at 1 wk postchallenge. Taken together, these results suggest that cDDR5-MAP immunization is an effective prophylactic vaccine strategy that suppresses and delays viral propagation during the initial HIV-1 transmission for the containment of HIV-1 replication subsequent to infection. PMID- 16365441 TI - Listeria monocytogenes activated p38 MAPK and induced IL-8 secretion in a nucleotide-binding oligomerization domain 1-dependent manner in endothelial cells. AB - Nucleotide-binding oligomerization domain (Nod) proteins serve as intracellular pattern recognition molecules recognizing peptidoglycans. To further examine intracellular immune recognition, we used Listeria monocytogenes as an organism particularly amenable for studying innate immunity to intracellular pathogens. In contrast to wild-type L. monocytogenes, the nonpathogenic Listeria innocua, or L. monocytogenes mutants lacking internalin B or listeriolysin O, poorly invaded host cells and escaped into host cell cytoplasm, respectively, and were therefore used as controls. In this study, we show that only the invasive wild-type L. monocytogenes, but not the listeriolysin O- or internalin B-negative L. monocytogenes mutants or L. innocua, substantially induced IL-8 production in HUVEC. RNA interference and Nod1-overexpression experiments demonstrated that Nod1 is critically involved in chemokine secretion and NF-kappaB activation initiated by L. monocytogenes in human endothelial cells. Moreover, we show for the first time that Nod1 mediated activation of p38 MAPK signaling induced by L. monocytogenes. Finally, L. monocytogenes- and Nod1-induced IL-8 production was blocked by a specific p38 inhibitor. In conclusion, L. monocytogenes induced a Nod1-dependent activation of p38 MAPK signaling and NF-kappaB which resulted in IL-8 production in endothelial cells. Thus, Nod1 is an important component of a cytoplasmic surveillance pathway. PMID- 16365440 TI - Helminth-primed dendritic cells alter the host response to enteric bacterial infection. AB - To examine whether intestinal helminth infection may be a risk factor for enteric bacterial infection, a murine model was established using the intestinal helminth Heligomosomoides polygyrus and a murine pathogen Citrobacter rodentium, which causes infectious colitis. Using this model we recently have shown that coinfection with the Th2-inducing H. polygyrus and C. rodentium promotes bacterial-associated disease and colitis. In this study, we expand our previous observations and examine the hypothesis that dendritic cells (DC) stimulated by helminth infection may play an important role in the regulation of the intestinal immune response to concurrent C. rodentium infection as well as in the modulation of the bacterial pathogenesis. We show that H. polygyrus infection induces DC activation and IL-10 expression, and that adoptive transfer of parasite-primed DC significantly impairs host protection to C. rodentium infection, resulting in an enhanced bacterial infection and in the development of a more severe colonic injury. Furthermore, we demonstrate that adoptive transfer of parasite-primed IL 10-deficient DCs fails to result in the development of a significantly enhanced C. rodentium-mediated colitis. Similarly, when the DC IL-10 response was neutralized by anti-IL-10 mAb treatment in mice that received parasite-primed DC, no deleterious effect of the parasite-primed DC on the host intestinal response to C. rodentium was detected. Thus, our results provide evidence to indicate that the H. polygyrus-dependent modulation of the host response to concurrent C. rodentium infection involves IL-10-producing DCs. PMID- 16365442 TI - Functional importance of regional differences in localized gene expression of receptors for IL-13 in murine gut. AB - IL-13 induces a STAT6-dependent hypercontractility of intestinal smooth muscle that is mediated by binding to the IL-13Ralpha1 component of the type 2 IL-4R that is linked to STAT6. IL-13 also binds to the IL-13Ralpha2 that is not linked to STAT6 and functions to limit the effects of IL-13 in vivo. In this study we assessed the contributions of regional and cellular differences in the distribution of the IL-13R components to the physiological regulation of smooth muscle function in wild-type mice and mice deficient in STAT6 or IL-13Ralpha2. The expression of IL-13 and IL-13Ralpha2 was higher in colon than in small intestine. Laser capture microdissection of specific cell types revealed that the expression of IL-13Ralpha2 was higher in the smooth muscle layer compared with levels in the epithelial cells of the mucosa. In contrast, there was a uniform distribution of IL-13alpha1 in smooth muscle, epithelia, and myenteric neurons. The significant hypercontractility of smooth muscle in mice deficient in IL 13Ralpha2, but not in STAT6, shows the physiological importance of IL-13 binding to IL-13Ralpha2. The pronounced differences in the expression of IL-13Ralpha2 suggest that the gut has developed sophisticated mechanisms for controlling the physiological and pathophysiological activities of IL-13. PMID- 16365443 TI - CD154 is essential for protective immunity in experimental salmonella infection: evidence for a dual role in innate and adaptive immune responses. AB - CD40-CD154 interactions are of central importance in the induction of humoral and cellular immune responses. In the present study, CD154-deficient (CD154-/-) mice were used to assess the role of CD40-CD154 interactions in regulating the immune response to a systemic Salmonella infection. Compared with C57BL/6 (CD154+/+) controls, CD154-/- mice were hypersusceptible to infection by an attenuated strain of Salmonella enterica serovar Typhimurium (S. typhimurium), as evidenced by decreased survival rate and mean time to death, which correlated with increased bacterial burden and persistence in target organs. CD154-/- mice exhibited a defect both in the production of IL-12, IFN-gamma, and NO during the acute phase of the disease and in the generation of Salmonella-specific Ab responses and Ig isotype switching. Furthermore, when CD154-/- animals were administered a sublethal dose of attenuated S. typhimurium and subsequently challenged with a virulent homologous strain, all mice succumbed to an overwhelming infection. Similar treatment of CD154+/+ mice consistently resulted in > or =90% protection. The lack of protective immunity in CD154-/- mice correlated with a decreased T cell recall response to Salmonella Ags. Significant protection against virulent challenge was conferred to presensitized CD154-/- mice by transfer of serum or T cells from immunized CD154+/+ mice. For best protection, however, a combination of immune serum and T cells was required. We conclude that intercellular communications via the CD40-CD154 pathway play a critical role in the induction of type 1 cytokine responses, memory T cell generation, Ab formation, and protection against primary as well as secondary Salmonella infections. PMID- 16365444 TI - IL-15 regulates CD8+ T cell contraction during primary infection. AB - During the course of acute infection with an intracellular pathogen, Ag-specific T cells proliferate in the expansion phase, and then most of the T cells die by apoptosis in the following contraction phase, but the few that survive become memory cells and persist for a long period of time. Although IL-15 is known to play an important role in long-term maintenance of memory CD8+ T cells, the potential roles of IL-15 in CD8+ T cell contraction are not known. Using an adoptive transfer system of OT-I cells expressing OVA257-264/Kb-specific TCR into control, IL-15 knockout (KO) and IL-15 transgenic (Tg) mice followed by challenge with recombinant Listeria monocytogenes expressing OVA, we found that the survival of CD44+CD62L-CD127- effector OT-I cells during the contraction phase is critically dependent on IL-15. In correlation with the expression level of Bcl-2 in OT-I cells, the number of OT-I cells was markedly reduced in IL-15 KO mice but remained at a high level in IL-15 Tg mice during the contraction phase, compared with control mice. In vivo administration of rIL-15 during the contraction phase in IL-15 KO mice inhibited the contraction of effector OT-I cells accompanied by up-regulation of Bcl-2 expression. Furthermore, enforced expression of Bcl-2 protected the majority of effector OT-I cells from death in IL-15 KO mice after infection. These results suggest that IL-15 plays a critical role in protecting effector CD8+ T cells from apoptosis during the contraction phase following a microbial infection via inducing antiapoptotic molecules. PMID- 16365445 TI - Neuronal nitric oxide synthase is necessary for elimination of Giardia lamblia infections in mice. AB - NO produced by inducible NO synthase (NOS2) is important for the control of numerous infections. In vitro, NO inhibits replication and differentiation of the intestinal protozoan parasite Giardia lamblia. However, the role of NO against this parasite has not been tested in vivo. IL-6-deficient mice fail to control Giardia infections, and these mice have reduced levels of NOS2 mRNA in the small intestine after infection compared with wild-type mice. However, NOS2 gene targeted mice and wild-type mice treated with the NOS2 inhibitor N-iminoethyl-L lysine eliminated parasites as well as control mice. In contrast, neuronal NOS (NOS1)-deficient mice and wild-type mice treated with the nonspecific NOS inhibitor NG-nitro-L-arginine methyl ester and the NOS1-specific inhibitor 7 nitroindazole all had delayed parasite clearance. Finally, Giardia infection increased gastrointestinal motility in wild-type mice, but not in SCID mice. Furthermore, treatment of wild-type mice with NG-nitro-L-arginine methyl ester or loperamide prevented both the increased motility and the elimination of parasites. Together, these data show that NOS1, but not NOS2, is necessary for clearance of Giardia infection. They also suggest that increased gastrointestinal motility contributes to elimination of the parasite and may also contribute to parasite-induced diarrhea. Importantly, this is the first example of NOS1 being involved in the elimination of an infection. PMID- 16365446 TI - Endotoxin-induced expression of murine bactericidal permeability/increasing protein is mediated exclusively by toll/IL-1 receptor domain-containing adaptor inducing IFN-beta-dependent pathways. AB - Antimicrobial effector proteins are a key mechanism for the innate immune system to combat pathogens once they infect the host. We report the identification and cloning of the mouse homologue of human bactericidal permeability/increasing protein (BPI). Mouse BPI is constitutively expressed in lymphatic organs and tissues as well as in mouse testis. Upon stimulation with different TLR ligands, mouse BPI is strongly expressed in granulocytes and, surprisingly, in bone marrow derived dendritic cells. Mouse BPI is most strongly induced by bacterial LPS through a signaling pathway that is completely dependent on TLR4-Toll/IL-1R domain-containing adaptor inducing IFN-beta. Functional studies revealed that mouse BPI does have the potential to neutralize LPS and inhibits bacterial growth. Mouse BPI is expressed in granulocytes and bone marrow-derived dendritic cells, and the transcriptional activation is controlled by TLRs. PMID- 16365447 TI - CD4+ T lymphocytes expressing CD40 ligand help the IgM antibody response to soluble pneumococcal polysaccharides via an intermediate cell type. AB - Streptococcus pneumoniae causes serious infections in children, the elderly, and immunocompromised patients. Protection against infections with S. pneumoniae is mediated through Abs against the capsular polysaccharides (caps-PS). We previously showed that the murine Ab response to caps-PS is dependent on CD40 CD40L interaction. In the present paper, we addressed the question of whether the CD40-CD40L-mediated modulation of the anti-caps-PS immune reaction is the result of a direct interaction between B lymphocytes and T lymphocytes or of an indirect interaction. SCID/SCID mice reconstituted with B lymphocytes from wild-type mice did not mount anti-caps-PS Abs. SCID/SCID mice reconstituted with B lymphocytes from wild-type mice and CD4+ T lymphocytes from wild-type mice but not CD4+ T lymphocytes from CD40L knockout mice stimulated the anti-caps-PS Ab response. This indicated that CD4+ T lymphocytes stimulated the anti-caps-PS Ab response in a CD40L-dependent manner. SCID/SCID mice reconstituted with B lymphocytes from CD40 knockout mice and CD4+ T lymphocytes from wild-type mice generated an anti caps-PS Ab response that could be inhibited by MR1, a blocking anti-CD40L Ab. These data indicated that CD4+ T lymphocytes stimulated the anti-caps-PS Ab response in an indirect way. Finally, lethally irradiated CD40 knockout mice reconstituted with bone marrow from wild-type mice mounted an anti-caps-PS Ab response that was comparable to the Ab response in wild-type mice, revealing that the required CD40 was on hemopoietic cells. In conclusion, we provide evidence that CD4+ T lymphocytes expressing CD40L stimulate the Ab response to soluble caps-PS by interacting with CD40-expressing non-B cells. PMID- 16365448 TI - Memory T cell populations in the lung airways are maintained by continual recruitment. AB - Effector memory T cell populations in the periphery play a key role in cellular immune responses to secondary infections. However, it is unclear how these populations are maintained under steady-state conditions in nonlymphoid peripheral sites, such as the lung airways. In this study, we show that LFA-1 expression is selectively down-regulated following entry of memory T cells into the lung airways. Using Sendai virus as a mouse model of respiratory virus infection, we use LFA-1 expression levels to demonstrate that effector memory T cell populations in the lung airways are maintained by continual recruitment of new cells from the circulation. The rate of memory cell recruitment is surprisingly rapid, resulting in replacement of 90% of the population every 10 days, and is maintained for well over 1 year following viral clearance. These data indicate that peripheral T cell memory is dynamic and depends on a systemic source of T cells. PMID- 16365449 TI - Identification and characterization of U83A viral chemokine, a broad and potent beta-chemokine agonist for human CCRs with unique selectivity and inhibition by spliced isoform. AB - Leukotropic human herpesvirus 6 (HHV-6) establishes a persistent infection associated with inflammatory diseases and encodes chemokines that could chemoattract leukocytes for infection or inflammation. HHV-6 variant A encodes a distant chemokine homolog, U83A, and a polymorphism promoting a secreted form was identified. U83A and three N-terminal modifications were expressed and purified, and activities were compared with a spliced truncated isoform, U83A-Npep. U83A efficiently and potently induced calcium mobilization in cells expressing single human CCR1, CCR4, CCR6, or CCR8, with EC50 values <10 nM. U83A also induced chemotaxis of Th2-like leukemic cells expressing CCR4 and CCR8. High-affinity binding, 0.4 nM, was demonstrated to CCR1 and CCR5 on monocytic/macrophage cells, and pretreatment with U83A or modified forms could block responses for endogenous ligands. U83A-Npep acted only as antagonist, efficiently blocking binding of CCL3 to CCR1 or CCR5 on differentiated monocytic/macrophage leukemic cells. Furthermore, CCL3 induction of calcium signaling via CCR1 and CCL1 induced chemotaxis via CCR8 in primary human leukocytes was inhibited. Thus, this blocking by the early expressed U83A-Npep could mediate immune evasion before finishing the replicative cycle. However, late in infection, when full-length U83A is made, chemoattraction of CCR1-, CCR4-, CCR5-, CCR6-, and CCR8-bearing monocytic/macrophage, dendritic, and T lymphocyte cells can facilitate dissemination via lytic and latent infection of these cells. This has further implications for neuroinflammatory diseases such as multiple sclerosis, where both cells bearing CCR1/CCR5 plus their ligands, as well as HHV-6A, have been linked. Applications also discussed include novel vaccines/immunotherapeutics for cancer and HIV as well as anti-inflammatories. PMID- 16365450 TI - Chemotactic responses of IL-4-, IL-10-, and IFN-gamma-producing CD4+ T cells depend on tissue origin and microbial stimulus. AB - Th1- and Th2-polarized immune responses are crucial in the defense against pathogens but can also promote autoimmunity and allergy. The chemokine receptors CXCR3 and CCR4 have been implicated in differential trafficking of IFN-gamma- and IL-4-producing T cells, respectively, but also in tissue and inflammation specific homing independent of cytokine responses. Here, we tested whether CD4+ T cells isolated from murine tissues under homeostatic or inflammatory conditions exhibit restricted patterns of chemotactic responses that correlate with their production of IFN-gamma, IL-4, or IL-10. In uninfected mice, IL-4-producing T cells preferentially migrated to the CCR4 ligand, CCL17, whereas IFN-gamma expressing T cells as well as populations of IL-4+ or IL-10+ T cells migrated to the CXCR3 ligand, CXCL9. All cytokine-producing T cell subsets strongly migrated to the CXCR4 ligand, CXCL12. We assessed chemotaxis of T cells isolated from mice infected with influenza A virus or the nematode Nippostrongylus brasiliensis, which induce a strong Th1 or Th2 response in the lung, respectively. Unexpectedly, the chemotactic responses of IL-4+ T cells and T cells expressing the immunosuppressive cytokine IL-10 were influenced not only by the strongly Th1 or Th2-polarized environments but also by their anatomical localization, i.e., lung or spleen. In contrast, IFN-gamma+ T cells exhibited robust chemotaxis toward CXCL9 and had the most consistent migration pattern in both infection models. The results support a model in which the trafficking responses of many effector and regulatory T cells are regulated as a function of the infectious and tissue environments. PMID- 16365451 TI - Doxycycline modulates nitric oxide production in murine lung epithelial cells. AB - Many effective therapeutic agents exhibit effects that are different from their intended primary mode of action. Antibiotics such as doxycycline and erythromycin A are no exception. They also display anti-inflammatory activity. Using LA4 murine lung alveolar epithelial cells, effects of doxycycline and erythromycin A on inducible NO synthase (iNOS) NO production as well as iNOS protein and mRNA production were investigated. Induction of iNOS was accomplished by treatment with cytomix (TNF-alpha, IL-1beta, and IFN-gamma each at 5 ng/ml). Production of NO or iNOS was not detected in controls with or without erythromycin A. In the presence of cytomix, erythromycin A did not decrease NO, nitrite, iNOS protein, or mRNA production. In contrast, doxycycline caused a dose-dependent decrease in NO, nitrite, iNOS protein, and mRNA production in cytomix-treated cells. Doxycycline at 30 mug/ml produced a 90% decrease in nitrite and NO production and a 52% decrease in iNOS mRNA transcription compared with cytomix treatment alone. Actinomycin D treatment suggests that doxycycline decreases stability of iNOS mRNA in cytomix-treated cells. To determine a mechanism for the decrease in iNOS expression, NF-kappaB and AP-1 transcription regulatory systems and p38 MAPK were examined. Doxycycline treatment gave no statistically significant change in NF kappaB activation but did decrease p38 MAPK protein in cytomix-treated cells by 50%, suggesting that p38 MAPK may be responsible for stabilization of iNOS mRNA. These results demonstrate that doxycycline decreases NO production from iNOS by destabilization of iNOS mRNA via decreased expression of p38 MAPK. PMID- 16365453 TI - Lipopolysaccharide induces cyclooxygenase-2 in intestinal epithelium via a noncanonical p38 MAPK pathway. AB - Necrotizing enterocolitis (NEC), a severe intestinal inflammation in neonates, occurs following bacterial colonization of the gut. LPS-induced production of inflammatory factors in immature enterocytes may be a factor in NEC. Previously, we described LPS-induced p38 MAPK-dependent expression of cyclooxygenase-2 (COX 2) in rat IEC-6 cells. In this study, we examine COX-2 expression in newborn rat intestinal epithelium and further characterize the mechanisms of COX-2 regulation in enterocytes. Induction of NEC by formula feeding/hypoxia increased phospho-p38 and COX-2 levels in the intestinal mucosa. Celecoxib, a selective COX-2 inhibitor, exacerbated the disease, suggesting a protective role for COX-2. COX-2 was induced in the intestinal epithelium by LPS in vivo and ex vivo. The latter response was attenuated by the p38 inhibitor SB202190, but not by inhibitors of ERK, JNK, or NF-kappaB. In IEC-6 enterocytes, COX-2 was induced by the expression of MAPK kinase 3 EE (MKK3EE), a constitutive activator of p38, but not of activators of ERK or JNK pathways. However, neither MKK3/6 nor MKK4, the known p38 upstream kinases, were activated by LPS. Dominant-negative MKK3 or MKK4 or SB202190 failed to prevent LPS-induced, p38-activating phosphorylation, ruling out important roles of these kinases or p38 autophosphorylation. LPS increased COX-2 and activating phosphorylation of p38 with similar dose-response. Blockade of LPS-induced expression of COX-2-luciferase reporter and destabilization of COX 2 message by SB202190 indicate that p38 regulates COX-2 at transcription and mRNA stability levels. Our data indicate that p38-mediated expression of COX-2 proceeds through a novel upstream pathway and support the role of the neonate's enterocytes as bacterial sensors. PMID- 16365452 TI - Lipoteichoic acid-induced nitric oxide production depends on the activation of platelet-activating factor receptor and Jak2. AB - NO production by macrophages in response to lipoteichoic acid (LTA) and a synthetic lipopeptide (Pam3CSK4) was investigated. LTA and Pam3CSK4 induced the production of both TNF-alpha and NO. Inhibitors of platelet-activating factor receptor (PAFR) blocked LTA- or Pam3CSK4-induced production of NO but not TNF alpha. Jak2 tyrosine kinase inhibition blocked LTA-induced production of NO but not TNF-alpha. PAFR inhibition blocked phosphorylation of Jak2 and STAT1, a key factor for expressing inducible NO synthase. In addition, LTA did not induce IFN beta expression, and p38 mitogen-activated protein serine kinase was necessary for LTA-induced NO production but not for TNF-alpha production. These findings suggest that Gram-positive bacteria induce NO production using a PAFR signaling pathway to activate STAT1 via Jak2. This PAFR/Jak2/STAT1 signaling pathway resembles the IFN-beta, type I IFNR/Jak/STAT1 pathway described for LPS. Consequently, Gram-positive and Gram-negative bacteria appear to have different but analogous mechanisms for NO production. PMID- 16365454 TI - Class IB-phosphatidylinositol 3-kinase (PI3K) deficiency ameliorates IA-PI3K induced systemic lupus but not T cell invasion. AB - Class I PI3K catalyzes formation of 3-poly-phosphoinositides. The family is divided into IA isoforms, activated by Tyr kinases and the IB isoform (PI3Kgamma), activated by G protein-coupled receptors. Mutations that affect PI3K are implicated in chronic inflammation, although the differential contribution of each isoform to pathology has not been elucidated. Enhanced activation of class IA-PI3K in T cells extends CD4+ memory cell survival, triggering an invasive lymphoproliferative disorder and systemic lupus. As both IA- and IB-PI3K isoforms regulate T cell activation, and activated pathogenic CD4+ memory cells are involved in triggering systemic lupus, we examined whether deletion of IB could reduce the pathological consequences of increased IA-PI3K activity. IB-PI3Kgamma deficiency did not abolish invasion or lymphoproliferation, but reduced CD4+ memory cell survival, autoantibody production, glomerulonephritis, and systemic lupus. Deletion of the IB-PI3Kgamma isoform thus decreased survival of pathogenic CD4+ memory cells, selectively inhibiting systemic lupus development. These results validate the PI3Kgamma isoform as a target for systemic lupus erythematosus treatment. PMID- 16365455 TI - Ligation of CD28 stimulates the formation of a multimeric signaling complex involving grb-2-associated binder 2 (gab2), SRC homology phosphatase-2, and phosphatidylinositol 3-kinase: evidence that negative regulation of CD28 signaling requires the gab2 pleckstrin homology domain. AB - Grb-2-associated binder (Gab)2 is a scaffolding adaptor protein that has been reported to promote growth factor and cytokine receptor signal transduction, but inhibit TCR-mediated signaling events. In this study, we show that ligation of CD28 by its natural ligand B7-1/CD80, induces tyrosine phosphorylation of Gab2 and its coassociation with Src homology phosphatase (SHP)-2 and class IA PI3K in Jurkat cells. Overexpression of wild-type Gab2 revealed a negative role in regulation of CD3/CD28 induction of the transcription factors NF-kappaB and AP-1. To characterize this inhibitory function further, we used Gab2 mutants unable to bind either PI3K or SHP-2 and a PH domain deletion mutant. Although PI3K has previously been implicated as necessary for Gab2-mediated inhibition of TCR signaling, Gab2 mutants defective in their ability to bind PI3K or SHP-2 retained their inhibitory function, whereas deletion of the PH domain ablated the inhibitory effect of Gab2. Together, these data demonstrate that CD28 stimulation of T cells is sufficient to induce an inhibitory multimeric signaling complex involving Gab2, SHP-2, and PI3K. Furthermore, the inhibitory capacity of Gab2 is strictly dependent upon the integrity of its PH domain, suggesting phosphoinositide-mediated membrane recruitment is important to Gab2 function in T cells. PMID- 16365456 TI - NF-kappaB and activator protein 1 response elements and the role of histone modifications in IL-1beta-induced TGF-beta1 gene transcription. AB - Abnormal expression of TGF-beta1 is believed to play an important role in the pathogenesis of a number of chronic inflammatory and immune lung diseases, including asthma, chronic obstructive pulmonary disease, and pulmonary fibrosis. Gene activation in eukaryotes requires coordinated use of specific cell signals, chromatin modifications, and chromatin remodeling. We studied the roles of the ubiquitous inflammatory transcription factors, NF-kappaB and AP-1, in activation of the TGF-beta1 gene and histone acetylation at the TGF-beta1 promoter. IL-1beta induced TGF-beta1 protein secretion and mRNA expression were prevented by actinomycin D and were attenuated by the inhibitor of kappaB kinase 2 inhibitor AS602868 and the JNK inhibitor SP600125, suggesting a degree of transcriptional regulation mediated by the NF-kappaB and AP-1 pathways. We demonstrated that IL 1beta activated the p65 subunit of NF-kappaB and the c-Jun subunit of AP-1. Using chromatin immunoprecipitation assays, we observed a sequential recruitment of p65 and c-Jun, accompanying ordered elevation of the levels of histone H4 and H3 acetylation and recruitment of RNA polymerase II at distinct regions in the native TGF-beta1 promoter. The specific NF-kappaB and AP-1 binding sites in the TGF-beta1 promoter were confirmed by an ELISA-based binding assay, and evidence for histone hyperacetylation in TGF-beta1 induction was supported by the observation that the histone deacetylase inhibitor trichostatin A enhanced basal and IL-1beta-induced TGF-beta1 mRNA expression. Our results suggest that IL-1beta stimulated transcription of TGF-beta1 is temporally regulated by NF-kappaB and AP 1 and involves histone hyperacetylation at distinct promoter sites. PMID- 16365457 TI - Molecular characterization of rat leukocyte P-selectin glycoprotein ligand-1 and effect of its blockade: protection from ischemia-reperfusion injury in liver transplantation. AB - P-selectin glycoprotein ligand-1 (PSGL-1) mediates the initial tethering of leukocytes to activated platelets and endothelium. We report molecular cloning and characterization of the rat PSGL-1 gene. A neutralizing Ab was generated, and its binding epitope was mapped to the N-terminal binding region of rat PSGL-1. We examined the effects of early PSGL-1 blockade in rat liver models of cold ischemia, followed by ex vivo reperfusion or transplantation (orthotopic liver transplantation (OLT)) using an anti-PSGL-1 Ab with diminished Fc-mediated effector function. In the ex vivo hepatic cold ischemia and reperfusion model, pretreatment with anti-PSGL-1 Ab improved portal venous flow, increased bile production, and decreased hepatocellular damage. Rat pretreatment with anti-PSGL 1 Ab prevented hepatic insult in a model of cold ischemia, followed by OLT, as assessed by 1) decreased hepatocellular damage (serum glutamic oxaloacetic transaminase/glutamic-pyruvic transaminase levels), and ameliorated histological features of ischemia/reperfusion injury, consistent with extended OLT survival; 2) reduced intrahepatic leukocyte infiltration, as evidenced by decreased expression of P-selectin, ED-1, CD3, and OX-62 cells; 3) inhibited expression of proinflammatory cytokine genes (TNF-alpha, IL-1beta, IL-6, IFN-gamma, and IL-2); and 4) prevented hepatic apoptosis accompanied by up-regulation of antiapoptotic Bcl-2/Bcl-xL protective genes. Thus, targeting PSGL-1 with a blocking Ab that has diminished Fc-mediated effector function is a simple and effective strategy that provides the rationale for novel therapeutic approaches to maximize the organ donor pool through the safer use of liver transplants despite prolonged periods of cold ischemia. PMID- 16365458 TI - Bacterial-responsive B lymphocytes induce periodontal bone resorption. AB - Host immune responses play a key role in periodontal diseases. We have found that B lymphocytes in human periodontal lesions bear abundant receptor activator of NF kappaB ligand (RANKL), a major factor in the regulation of osteoclast differentiation. The purpose of this study was to evaluate Actinobacillus actinomycetemcomitans-responsive B lymphocytes in their level of RANKL expression and their effects on periodontal bone resorption. Congenitally athymic Rowett rats received injections of formalin-fixed A. actinomycetemcomitans into the gingival papillae, and donor B cells from normal rats immunized with A. actinomycetemcomitans were transferred via tail vein injection. We demonstrated that B cells from A. actinomycetemcomitans-immunized animals had greater levels of RANKL expression and induced a significantly higher level of osteoclast differentiation from RAW 264.7 cells than did nonimmune B cells that were not Ag specific. This activity was eliminated by incubation with the RANKL decoy receptor osteoprotegerin fusion protein. A. actinomycetemcomitans-binding B cell (ABB) and RANKL-expressing B cells were recovered from the gingival tissues of recipient rats transferred with ABB, but not from recipients of PBS nonimmune B cells or A. actinomycetemcomitans nonbinding B cells. Also, recipients of ABB exhibited increased osteoclast formation on the alveolar bone surface and significant periodontal bone resorption. This effect was antagonized by injection of osteoprotegerin fusion protein into the local gingival tissues. In summary, this study suggests that B lymphocytes can contribute to increased periodontal bone resorption in the absence of T lymphocytes. This effect is associated with the up-regulation of RANKL expression. PMID- 16365459 TI - Innate stimuli accentuate end-organ damage by nephrotoxic antibodies via Fc receptor and TLR stimulation and IL-1/TNF-alpha production. AB - Innate stimuli are well recognized as adjuvants of the systemic immune response. However, their role in driving end-organ disease is less well understood. Whereas the passive transfer of glomerular-targeting Abs alone elicited minimal renal disease, the concomitant delivery of innate stimuli triggered severe nephritis, characterized by proliferative glomerulonephritis with crescent formation, and tubulointerstitial disease. Specifically, stimulating TLR2, TLR3, TLR4, and TLR5 by using peptidoglycan, poly(I:C), LPS, and flagellin, respectively, all could facilitate anti-glomerular Ab-elicited nephritis. In this model, innate and immune triggers synergistically activated several cytokines and chemokines, including IL-1, IL-6, TNF-alpha, and MCP-1, some of which were demonstrated to be absolutely essential for the development of renal disease. Genetic studies revealed that, whereas the innate trigger is dependent on TLR/IL-1R-associated kinase-mediated signaling, the immune component was contingent on FcR-mediated signals. Importantly, infiltrating leukocytes as well as intrinsic glomerular cells may both serve to integrate these diverse signals. Extrapolating to spontaneous immune-mediated nephritis, although the adaptive immune system may be important in generating end-organ targeting Abs, the extent of damage inflicted by these Abs may be heavily dependent on cues from the innate immune system. PMID- 16365461 TI - SC5 mAb represents a unique tool for the detection of extracellular vimentin as a specific marker of Sezary cells. AB - Circulating malignant Sezary lymphocytes result from a clonal proliferation of memory/activated CD4(+)CD45RO(+) T lymphocytes primarily involving the skin. Recently, the CD158k/KIR3DL2 cell surface receptor has been identified to phenotypically characterize these cells. We previously described a mAb termed SC5 that identifies an unknown early activation cell membrane molecule. It is expressed selectively by T lymphocytes isolated from healthy individuals upon activation, and by circulating Sezary syndrome lymphocytes. In addition, we found that SC5 mAb was reactive with all resting T lymphocytes once permeabilized, indicating that SC5 mAb-reactive molecule might present distinct cellular localization according to the T cell activation status. In this study, we show for the first time that SC5 mAb recognizes the intermediate filament protein vimentin when exported to the extracellular side of the plasma membrane of viable Sezary malignant cells. We demonstrate that SC5 mAb is unique as it reacts with both viable malignant lymphocytes and apoptotic T cells. As vimentin is also detected rapidly at the cell membrane surface after normal T lymphocyte activation, it suggests that its extracellular detection on Sezary cells could be a consequence of their constitutive activation status. Finally, as a probable outcome of vimentin cell surface expression, autoantibodies against vimentin were found in the sera of Sezary syndrome patients. PMID- 16365460 TI - Azathioprine suppresses ezrin-radixin-moesin-dependent T cell-APC conjugation through inhibition of Vav guanosine exchange activity on Rac proteins. AB - We have shown recently that the azathioprine metabolite 6-Thio-GTP causes immunosuppression by blockade of GTPase activation in T lymphocytes. In the present study, we describe a new molecular mechanism by which 6-Thio-GTP blocks GTPase activation. Although 6-Thio-GTP could bind to various small GTPases, it specifically blocked activation of Rac1 and Rac2 but not of closely related Rho family members such as Cdc42 and RhoA in primary T cells upon stimulation with alphaCD28 or fibronectin. Binding of 6-Thio-GTP to Rac1 did not suppress Rac effector coupling directly but blocked Vav1 exchange activity upon 6-Thio-GTP hydrolysis, suggesting that 6-Thio-GTP loading leads to accumulation of 6-Thio GDP-loaded, inactive Rac proteins over time by inhibiting Vav activity. In the absence of apoptosis, blockade of Vav-mediated Rac1 activation led to a blockade of ezrin-radixin-moesin dephosphorylation in primary T cells and suppression of T cell-APC conjugation. Azathioprine-generated 6-Thio-GTP thus prevents the development of an effective immune response via blockade of Vav activity on Rac proteins. These findings provide novel insights into the immunosuppressive effects of azathioprine and suggest that antagonists of the Vav-Rac signaling pathway may be useful for suppression of T cell-dependent pathogenic immune responses. PMID- 16365462 TI - Models of systemic lupus erythematosus: development of autoimmunity following peptide immunizations of noninbred pedigreed rabbits. AB - Reported in this study are the initial results from studies to develop rabbit models of systemic lupus erythematosus (SLE) by immunizations using two distinct peptides on branched polylysine backbones (multiple Ag peptide)-peptides. Eleven rabbits received a peptide from the Sm B/B' spliceosomal complex previously shown to be immunogenic in rabbits, and 13 rabbits received a peptide from the rabbit N methyl-d-aspartate receptor NR2b. All 24 animals in different generations of pedigreed, noninbred rabbits produced peptide-specific responses. Anti-nuclear autoantibody responses, including anti-dsDNA, were seen in 17 of 24 rabbits. To date, two rabbits have been observed to have seizure-like events and a third nystagmus. A model for eliciting development of SLE in genetically related yet heterogeneous rabbits may more closely resemble development of human SLE than do some models in inbred mice. Through selective breeding, it may also ultimately provide additional information about the genetics and etiology of SLE and serve as a model for assessing new treatment options. PMID- 16365464 TI - Summaries for patients. Comparison of yoga, exercise, and education for the treatment of chronic low back pain. PMID- 16365463 TI - Targeted expression of the human thyrotropin receptor A-subunit to the mouse thyroid: insight into overcoming the lack of response to A-subunit adenovirus immunization. AB - The thyrotropin receptor (TSHR), the major autoantigen in Graves' disease, is posttranslationally modified by intramolecular cleavage to form disulfide-linked A- and B-subunits. Because Graves' hyperthyroidism is preferentially induced in BALB/c mice using adenovirus encoding the free A-subunit rather than full-length human TSHR, the shed A-subunit appears to drive the disease-associated autoimmune response. To further investigate this phenomenon, we generated transgenic mice with the human A-subunit targeted to the thyroid. Founder transgenic mice had normal thyroid function and were backcrossed to BALB/c. The A-subunit mRNA expression was confirmed in thyroid tissue. Unlike wild-type littermates, transgenic mice immunized with low-dose A-subunit adenovirus failed to develop TSHR Abs, hyperthyroidism, or splenocyte responses to TSHR Ag. Conventional immunization with A-subunit protein and adjuvants induced TSHR Abs lacking the characteristics of human autoantibodies. Unresponsiveness was partially overcome using high-dose, full-length human TSHR adenovirus. Although of low titer, these induced Abs recognized the N terminus of the A-subunit, and splenocytes responded to A-subunit peptides. Therefore, "non-self" regions in the B-subunit did not contribute to inducing responses. Indeed, transgenic mice immunized with high dose A-subunit adenovirus developed TSHR Abs with thyrotropin-binding inhibitory activity, although at lower titers than wild-type littermates, suggesting down regulation in the transgenic mice. In conclusion, in mice expressing a human A subunit transgene in the thyroid, non-self human B-subunit epitopes are not necessary to induce responses to the A-subunit. Our findings raise the possibility that autoimmunity to the TSHR in humans may not involve epitopes on a cross-reacting protein, but rather, strong adjuvant signals provided in bystander immune responses. PMID- 16365465 TI - Summaries for patients. Comparison of 3 strategies to improve the care of patients with pneumonia treated in the emergency department. PMID- 16365466 TI - Comparing yoga, exercise, and a self-care book for chronic low back pain: a randomized, controlled trial. AB - BACKGROUND: Chronic low back pain is a common problem that has only modestly effective treatment options. OBJECTIVE: To determine whether yoga is more effective than conventional therapeutic exercise or a self-care book for patients with chronic low back pain. DESIGN: Randomized, controlled trial. SETTING: A nonprofit, integrated health care system. PATIENTS: 101 adults with chronic low back pain. INTERVENTION: 12-week sessions of yoga or conventional therapeutic exercise classes or a self-care book. MEASUREMENTS: Primary outcomes were back related functional status (modified 24-point Roland Disability Scale) and "bothersomeness" of pain (11-point numerical scale). The primary time point was 12 weeks. Clinically significant change was considered to be 2.5 points on the functional status scale and 1.5 points on the bothersomeness scale. Secondary outcomes were days of restricted activity, general health status, and medication use. RESULTS: After adjustment for baseline values, back-related function in the yoga group was superior to the book and exercise groups at 12 weeks (yoga vs. book: mean difference, -3.4 [95% CI, -5.1 to - 1.6] [P < 0.001]; yoga vs. exercise: mean difference, -1.8 [CI, -3.5 to - 0.1] [P = 0.034]). No significant differences in symptom bothersomeness were found between any 2 groups at 12 weeks; at 26 weeks, the yoga group was superior to the book group with respect to this measure (mean difference, -2.2 [CI, -3.2 to - 1.2]; P < 0.001). At 26 weeks, back-related function in the yoga group was superior to the book group (mean difference, -3.6 [CI, -5.4 to - 1.8]; P < 0.001). LIMITATIONS: Participants in this study were followed for only 26 weeks after randomization. Only 1 instructor delivered each intervention. CONCLUSIONS: Yoga was more effective than a self care book for improving function and reducing chronic low back pain, and the benefits persisted for at least several months. PMID- 16365467 TI - Empirical anti-Candida therapy among selected patients in the intensive care unit: a cost-effectiveness analysis. AB - BACKGROUND: Mortality from invasive candidiasis is high. Low culture sensitivity and treatment delay contribute to increased mortality, but nonselective early therapy may result in excess costs and drug resistance. OBJECTIVE: To determine the cost-effectiveness of anti-Candida strategies for high-risk patients in the intensive care unit (ICU). DESIGN: Cost-effectiveness decision model. DATA SOURCES: Published data to 10 May 2005, identified from MEDLINE and Cochrane Library searches, ICU databases, expert estimates, and actual hospital costs. TARGET POPULATION: Patients in the ICU with suspected infection who have not responded to antibacterial therapy. TIME HORIZON: Lifetime. PERSPECTIVE: Societal. INTERVENTIONS: Fluconazole, caspofungin, amphotericin B, or lipid formulation of amphotericin B given as either empirical or culture-based therapy and no anti-Candida therapy. OUTCOME MEASURES: Incremental life expectancy and incremental cost per discounted life-year (DLY) saved. RESULTS OF BASE-CASE ANALYSIS: Ten percent of the target population will have invasive candidiasis. Empirical caspofungin therapy is the most effective strategy but is expensive (295,115 dollars per DLY saved). Empirical fluconazole therapy is the most reasonable strategy (12,593 dollars per DLY saved) and decreases mortality from 44.0% to 30.4% in patients with invasive candidiasis and from 22.4% to 21.0% in the overall target cohort. RESULTS OF SENSITIVITY ANALYSIS: Empirical fluconazole therapy is reasonable for likelihoods of invasive candidiasis greater than 2.5% or fluconazole resistance less than 24.0%. For higher resistance levels, empirical caspofungin therapy is preferred. For low prevalences of invasive candidiasis, culture-based fluconazole is reasonable. For prevalences exceeding 60%, empirical caspofungin therapy is reasonable. For caspofungin to be reasonable at a prevalence of 10%, its cost must be reduced by 58%. LIMITATIONS: Less severe illness and limited use of broad-spectrum antimicrobial agents, typical of smaller hospitals, could result in a lower risk for invasive candidiasis. CONCLUSIONS: In patients in the ICU with suspected infection who have not responded to antibiotic treatment, empirical fluconazole should reduce mortality at an acceptable cost. The use of empirical strategies in low-risk patients is not justified. PMID- 16365468 TI - Meta-analysis: the efficacy of strategies to prevent organ disease by cytomegalovirus in solid organ transplant recipients. AB - BACKGROUND: Cytomegalovirus (CMV), the most common opportunistic viral infection in solid organ transplant recipients, is associated with substantial morbidity and mortality. PURPOSE: To assess the efficacy of universal prophylaxis and preemptive approaches in preventing CMV organ disease and other complications in solid organ transplant recipients. DATA SOURCES: Using no language restrictions, the authors searched the following databases from their inception through May 2005: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, and Evidence-Based Medicine. The authors also reviewed abstracts from scientific meetings and contacted experts and pharmaceutical companies. STUDY SELECTION: Randomized, controlled trials that evaluated antiviral strategies for preventing CMV and associated complications in solid organ transplant recipients were selected. DATA EXTRACTION: Two reviewers independently extracted and assessed the data. DATA SYNTHESIS: The authors found 17 trials involving 1980 patients. Compared with placebo or no therapy, both universal prophylaxis (odds ratio [OR], 0.20 [95% CI, 0.13 to 0.31]) and preemptive strategies (OR, 0.28 [CI, 0.11 to 0.69]) reduced CMV organ disease. However, only universal prophylaxis seemingly reduced CMV organ disease in subgroups of patients at highest risk (donors with positive CMV serostatus and recipients with negative CMV serostatus and induction with antibodies). Both strategies reduced the rate of allograft rejection. Only universal prophylaxis statistically significantly reduced bacterial and fungal infections (OR, 0.49 [CI, 0.36 to 0.67]) and death (OR, 0.62 [CI, 0.40 to 0.96]). Both acyclovir and ganciclovir statistically significantly prevented CMV organ disease in the universal prophylaxis trials. LIMITATIONS: Studies were of modest size, and few studies assessed all outcomes. Studies did not always provide data that discriminated between CMV organ disease and CMV syndrome or data about the timing of CMV organ disease. CONCLUSIONS: Universal prophylaxis and preemptive strategies are beneficial in preventing CMV organ disease in solid organ transplant recipients. Both strategies are associated with a reduction in allograft rejection, but current data suggest that only universal prophylaxis reduces bacterial and fungal infections and death. Acyclovir and ganciclovir are both effective for universal prophylaxis. PMID- 16365469 TI - Effect of increasing the intensity of implementing pneumonia guidelines: a randomized, controlled trial. AB - BACKGROUND: Despite the development of evidence-based pneumonia guidelines, limited data exist on the most effective means to implement guideline recommendations into clinical practice. OBJECTIVE: To compare the effectiveness and safety of 3 guideline implementation strategies. DESIGN: Cluster-randomized, controlled trial. SETTING: 32 emergency departments in Pennsylvania and Connecticut. PATIENTS: 3219 patients with a clinical and radiographic diagnosis of pneumonia. INTERVENTIONS: The authors implemented a project-developed guideline for the initial site of treatment based on the Pneumonia Severity Index and performance of evidence-based processes of care at the emergency department level. Guideline implementation strategies were defined as low (n = 8), moderate (n = 12), and high intensity (n = 12). MEASUREMENTS: Effectiveness outcomes were the rate at which low-risk patients were treated on an outpatient basis and the performance of recommended processes of care. Safety outcomes included death, subsequent hospitalization for outpatients, and medical complications for inpatients. RESULTS: More low-risk patients (n = 1901) were treated as outpatients in the moderate-intensity and high-intensity groups than in the low intensity group (high-intensity group, 61.9%; moderate-intensity group, 61.0%; low-intensity group, 37.5%; P = 0.004). More outpatients (n = 1125) in the high intensity group received all 4 recommended processes of care (high-intensity group, 60.9%; moderate-intensity group, 28.3%; low-intensity group, 25.3%; P < 0.001); more inpatients (n = 2076) in the high-intensity group received all 4 recommended processes of care (high-intensity group, 44.3%; moderate-intensity group, 30.1%; low-intensity group, 23.0%; P < 0.001). No statistically significant differences in safety outcomes were observed across interventions. LIMITATIONS: Twenty percent of eligible patients were not enrolled, and data on effectiveness outcomes were not collected before the trial. CONCLUSIONS: Both moderate-intensity and high-intensity guideline implementation strategies safely increased the proportion of low-risk patients with pneumonia who were treated as outpatients. The high-intensity strategy was most effective for increasing the performance of the recommended processes of care for outpatients and inpatients. PMID- 16365470 TI - New concepts in the pathophysiology of inflammatory bowel disease. PMID- 16365471 TI - Letter from New Orleans. PMID- 16365472 TI - The discovery of insulin: the Rochester, New York, connection. AB - The discovery of insulin in Toronto by Dr. Frederick G. Banting and colleagues has been well chronicled. The story of how insulin therapy was introduced into the United States has been less detailed. The first patient to be treated with insulin in the United States resided in Rochester, New York, a city with a then newly developed medical school that had also tried to recruit Dr. Banting. A series of letters from that period provides a description of the course of a juvenile patient with diabetes before and after the use of insulin as a therapeutic agent. PMID- 16365473 TI - Controlling the troll: management of cytomegalovirus infection after transplantation. PMID- 16365474 TI - The man with no heart. PMID- 16365475 TI - Screening for HIV. PMID- 16365476 TI - Unintended consequences. PMID- 16365477 TI - Unintended consequences. PMID- 16365478 TI - Tumor necrosis factor-alpha inhibitor therapy in erosive polyarthritis secondary to systemic sclerosis. PMID- 16365479 TI - Medicare coverage of angiotensin-converting enzyme inhibitors. PMID- 16365480 TI - Effect of an unstirred layer on the membrane permeability of anandamide. AB - To study the effect of an unstirred layer (UL), we have investigated the exchange efflux kinetics of anandamide at 0 degrees C, pH 7.3, from albumin-free as well as from albumin-filled human red blood cell ghosts to media of various BSA concentrations ([BSA](o)). The rate constant (k(m)) of unidirectional flux from the outer membrane leaflet to BSA in the medium increased with the square root of [BSA](o) in accordance with the existence of a UL, which is a water layer adjacent to the membrane that is not subject to the same gross mixing that takes place in the rest of the medium. From k(m), it is possible to calculate the rate constant of anandamide dissociation from BSA (k(1)) if we know the membrane binding of anandamide, the equilibrium dissociation constant of BSA-anandamide complexes, and the diffusion constant of anandamide. We estimated k(1) to be 3.33 +/- 0.27 s(-1). The net flux of [(3)H]anandamide is balanced by an equal and opposite movement of nonradioactive anandamide in exchange efflux experiments. This means that our results are also valid for uptake. We show that for anandamide with rapid membrane translocation, UL causes a significant resistance to cellular uptake. Depicting the rate of anandamide uptake as a function of equilibrium water phase concentrations results in a parabolic uptake dependence. Such apparent "saturation kinetics" is often interpreted as indicating the involvement of transport proteins. The validity of such an interpretation is discussed. PMID- 16365481 TI - Psychiatric genetics--the new era: genetic research and some clinical implications. AB - Impressive advances in the last decade have been made in the genetics and neuroscience of neuropsychiatric illness. Synergies between complex genetics, elaboration of intermediate phenotypes (Egan et al. (2004) Schizophrenia. London: Blackwell) and novel applications in neuroimaging (Bookheimer et al. (2000) N Engl J Med, 343, 450-456) are revealing the effects of positively associated disease alleles on aspects of neurological function. Genes such as NRG-1, DISC1, RGS4, COMT, PRODH, DTNBP1, G72, DAAO, GRM3 (Harrison and Weinberger (2005) Mol Psychiatry, 10, 40-68) and others have been implicated in schizophrenia along with 5-HTTPR (Ogilvie et al. (1996) Lancet, 347, 731-733; Caspi et al. (2003) Science, 301, 386-389) and BDNF (Geller et al. (2004) Am J Psychiatry, 161, 1698 1700) in affective disorders. As the genetics and complex neurocircuits of these and disorders are being untangled, parallel applications in pharmacogenomics and gene-based drug metabolism are shaping a drive for personalized medicine. Genetic research and pharmacogenomics suggest that the subcategorization of individuals based on various sets of susceptibility alleles will make the treatment of neuropsychiatric and other illnesses more predictable and effective. PMID- 16365482 TI - Surgery for wrist fractures in the elderly: for better form or better function? PMID- 16365483 TI - Fusion rate of anterior cervical plating after corpectomy. AB - PURPOSE: To evaluate the neurological recovery and fusion rate of patients with myelopathy who were treated with anterior corpectomy and anterior cervical plating. METHODS: The results of 17 cervical myelopathy patients who underwent decompression and anterior cervical plating were retrospectively reviewed at a mean follow-up of 2 years. RESULTS: By Kurokawa score, 82.4% of patients showed excellent-to-good results. The fusion rates of 2-level and 3-level anterior cervical corpectomy, and of anterior plate fixation were 100%. There were no implant- or graft-related complications. Transient dysphagia in 9 (52.9%) patients resolved after a mean of 3 months (range, 1-9 months). CONCLUSION: The use of anterior cervical plating after anterior corpectomy and fusion with autologous bone graft greatly enhances arthrodesis. The improved fusion rate and low complication rate associated with anterior cervical plating may justify its use in the treatment of cervical spondylotic myelopathy. PMID- 16365484 TI - Magnetic resonance imaging study of the craniocervical junction. AB - PURPOSE: To confirm the accuracy of the standard parameters of Chamberlain's line, McGregor's line, McRae's line, and Ranawat's line in the diagnosis of basilar impression in a normal Asian population using magnetic resonance imaging (MRI) scans of the craniocervical junction, and to evaluate the use of MRI in the initial diagnosis of basilar impression. METHODS: We retrospectively reviewed MRI scans of the craniocervical junction in 114 Thai patients (61 men and 53 women) aged 20 to 89 years to assess the lines of Chamberlain, McGregor, McRae, and Ranawat. Patients with injury, tumour, or metastasis in the craniocervial junction; congenital anomalies; or clinical symptoms and signs of basilar impression were excluded. RESULTS: 39 of the 114 patients had the odontoid tip extended above Chamberlain's line by 1 mm to 9 mm (mean, 2.89 mm). Only 7 men and 5 women had the tip of the odontoid process extended beyond the normal range of 5 mm. The mean distance of the odontoid tip extended past McGregor's line in male and female subjects was 0.33 mm (standard deviation [SD], 1.2 mm) and 0.17 mm (SD, 0.7 mm), respectively, markedly less than those reported previously, although one male patient's odontoid tip extended 8 mm above the McGregor's line. Regarding McRae's line, no patient had the odontoid tip extended above the foremen magnum. The mean Ranawat's line in male and female subjects was 15.75 mm (range, 12-19; SD, 1.71 mm) and 14.09 mm (range, 11-17; SD, 1.38 mm), respectively. CONCLUSION: After using MRI to confirm the standard parameters of basilar impression as previously formulated from lateral skull radiographs, we recommend using McRae's line and Ranawat's line in the initial screening for basilar impression for the Asian population. Chamberlain's line and McGregor's line are viable alternatives, but clinicians must be aware of the potential for false positives. PMID- 16365485 TI - Management of malignant atlanto-axial tumours. AB - PURPOSE: To review the management and outcomes of patients with malignant tumours of the atlanto-axial spine treated over a 4-year period at a tertiary hospital. METHODS: A total of 12 patients (9 female and 3 male) with a mean age of 56.5 years (range, 35-71 years) were consecutively enrolled. At the time of presentation, 7 patients had received treatment for a known primary malignancy of the lung (n=4), colon (n=1), nasopharynx (n=1), or breast (n=1). The atlanto axial spine was the first presentation of malignant disease in the remaining 5 patients, whose subsequent diagnoses were multiple myeloma (n=1), carcinoma of the thyroid (n=1), malignant fibrous histiocytoma (n=2), and adenocarcinoma of unknown origin (n=1). Preoperative, postoperative, and follow-up radiographs and computed tomographic and magnetic resonance imaging scans were analysed with regard to the extent of tumour involvement, other spinal metastases, instability, and evidence of cord compression. Surgeries were performed for surgically fit patients with a life expectancy of more than 3 months. RESULTS: All patients presented with severe suboccipital neck pain. Two had mild neurological deficits (Frankel grade D). Two patients were treated conservatively with rigid neck collars and survived for 3 and 11 months. Surgical stabilisation with posterior instrumentation was performed in 10 patients. One patient received additional anterior surgery for tumour debulking and bone grafting. No neurological complications were recorded in the series. Excellent relief of pain was achieved, and the mean visual analogue scale pain score of the operated patients improved from 9.3 to 1.9. The mean survival of 9 patients after surgery was 13.1 months, while one patient was still living at the time of review (at postoperative 11 months). CONCLUSION: Surgical stabilisation in selected patients provided pain relief and preserved ambulatory status before the deterioration of overall function. PMID- 16365486 TI - Pyogenic vertebral osteomyelitis: a review of 14 cases. AB - PURPOSE: To review and evaluate the management of 14 patients with pyogenic vertebral osteomyelitis at National University Hospital, Singapore between 1998 and 2001. METHODS: Demographic pattern, predisposing factors, clinical presentation, co-morbidities, microbiology, treatment, and complications of 14 patients were retrospectively reviewed. RESULTS: The mean age at presentation was 62.5 years and the male to female ratio was 6:1. The mean follow-up duration was 12.5 months. The most common predisposing condition was diabetes mellitus (n=5). The most common site of infection was the lumbar spine (n=8), followed by the thoracic (n=4) and cervical (n=2) spine. Staphylococcus aureus was the most common causative organism isolated (n=9), followed by methicillin-resistant S aureus (n=3), Pseudomonas pseudomallei (n=1), and Streptococcus agalactiae (n=1). 12 patients were treated surgically: 8 by an anterior approach, 3 by a posterior approach, and one by a combination of the 2. The mean period of antibiotic use was 11.4 weeks. One patient with melioidosis involving the T9 to T11 vertebrae caused by P pseudomallei died of empyema and septicaemia 22 months after presentation. CONCLUSION: Pyogenic vertebral osteomyelitis is not uncommon in the elderly, especially those with predisposing conditions such as diabetes mellitus. Computed tomography-guided needle biopsy is recommended to investigate causative microorganisms. Aggressive surgical debridement and prolonged antibiotic therapy were necessary in patients with methicillin-resistant S aureus, P pseudomallei, and S agalactiae. PMID- 16365487 TI - Clinical results of modified Mitchell's osteotomy for hallux valgus augmented with oblique lesser metatarsal osteotomy. AB - PURPOSE: To evaluate postoperative results of modified Mitchell's osteotomy and its combination with oblique metatarsal osteotomy for the treatment of hallux valgus. METHODS: A total of 93 feet of 53 patients (2 men and 51 women) with hallux valgus underwent modified Mitchell's osteotomy and were followed up for at least 5 years. Patients' age ranged from 17 to 83 years, and the duration of follow-up ranged from 5 years one month to 18 years 4 months. Modified Mitchell's osteotomy was performed on 53 feet in 31 patients (group A), whereas modified Mitchell's osteotomy augmented with oblique lesser metatarsal osteotomy was performed to the remaining 40 feet in 22 patients (group B). Postoperative results were assessed using a clinical assessment system developed by the Tokyo Medical University based on 5 categories: pain in the first metatarsophalangeal, deformity of the metatarsophalangeal, plantar callosity and/or metatarsalgia of lesser metatarsals, the use of commercially available shoes, and local inflammatory symptoms. RESULTS: Mean total score improved from 3.8 to 7.9 on a 10 point scale. Scores for 2 categories--plantar callosity and/or metatarsalgia and the use commercially available shoes--were significantly higher in group B at postoperative 5 years. Before surgery, at postoperative 3 weeks, and at postoperative 5 years, respectively, the mean hallux valgus angles were 34.2, 12.0, and 17.1 degrees; mean M1-M2 angles were 16.7, 7.4, and 8.7 degrees; mean M1-M5 angles were 34.9, 25.8, and 26.6 degrees; and mean sesamoid bone shifts were 8.7 mm, 4.3 mm, and 5.9 mm. CONCLUSION: Modified Mitchell's osteotomy shortens the length of the first metatarsal bone and thus relieves tension in soft tissues such as the adductor hallucis. Nonetheless, the procedure can induce metatarsophalangeal joint malalignment and metatarsalgia, and plantar callosity may develop or persist after surgery. Combining oblique metatarsal osteotomy of the lesser metatarsal bones is useful in patients with uneven metatarsal bone lengths and metatarsophalangeal joint malalignment. PMID- 16365488 TI - Radiographic assessment of congenital talipes equinovarus: strapping versus forced dorsiflexion. AB - PURPOSE: To compare 2 radiographic assessment methods for congenital talipes equinovarus: strapping the ankle with tapes versus dorsiflexion with a wooden block. METHODS: Anteroposterior and lateral radiographs were taken with the ankle strapped by tapes or dorsiflexed by a wooden block. Talocalcaneal and tibiocalcaneal angles were measured and analysed. RESULTS: 20 radiographic assessments were performed on 14 patients. From the anteroposterior view, the mean talocalcaneal angle with strapping was 19.4 degrees (range, 0-34.0 degrees; standard deviation [SD], 11.2 degrees). From the lateral view, the mean talocalcaneal angle with strapping was 22.0 degrees (range, 8.0-38.8 degrees; SD, 8.9 degrees) and with forced dorsiflexion was 21.0 degrees (range, 0-52.7 degrees; SD, 11.3 degrees), with a mean absolute difference of 9.4 degrees between the 2 groups. The mean tibiocalcaneal angle with strapping was 77.9 degrees (range, 43.2-120.0 degrees; SD, 18.8 degrees) and with forced dorsiflexion was 78.2 degrees (range, 29.5-134.0 degrees; SD, 24.7 degrees), with a mean absolute difference of 15.7 degrees between the 2 groups. The difference between the tibiocalcaneal measurements in the 2 groups was statistically significant (p=0.026). CONCLUSION: Similar results were obtained for talocalcaneal and tibiocalcaneal angles measured with strapping and forced dorsiflexion. However, the strapping method provides better stabilisation while radiographs are taken. PMID- 16365489 TI - Dall-Miles cable and plate fixation system in the treatment of periprosthetic femoral fractures: a review of 20 cases. AB - PURPOSE: To assess the effectiveness of Dall-Miles cables and plates in the treatment of periprosthetic femoral fractures around hip arthroplasties. METHODS: We retrospectively reviewed 20 cases of periprosthetic femoral fracture treated with the Dall-Miles cable and plate system between November 1999 and December 2002 in James Paget Hospital in the United Kingdom. Demographic data, fracture type, and surgical outcome of the patients were reviewed and assessed. RESULTS: Of the 20 cases of periprosthetic femoral fracture, one was of Vancouver type A, 15 were of type B1, and 4 were of type C. The mean age at operation was 78.5 years (range, 68-90 years) with a male to female ratio of 11:9. The mean time to union was 3.9 months (range, 3-8 months), and the mean follow-up period was 19.4 months (range, 1-4 years). 15 patients achieved satisfactory results, including 2 delayed unions that healed with a residual varus deformity of 15 degrees in the femoral component. Three patients developed deep-seated wound infections. No nonunion of fractures or fixation system failures occurred. CONCLUSION: The Dall Miles cable and plate fixation system alone is a sufficient treatment for most periprosthetic femoral fractures. In the management of B1 fractures, there was no significant difference in clinical outcome, despite the various combinations of cables and screws used for stabilisation of the plate. However, in cases of comminution at the fracture site, junctional grafting with morsellised autograft may be used in addition to internal fixation to accelerate union. PMID- 16365490 TI - Schwannomas of the hand and wrist: long-term results and review of the literature. AB - PURPOSE: To review the long-term results of 14 cases of histopathologically confirmed schwannoma of the hand and wrist treated with intracapsular tumour removal. METHODS: All operations were performed under loupe magnification and microsurgical dissection, and the tumour was removed after capsular (including endoneurium) incision. Patients were assessed for any residual symptoms. Sensory testing of light touch and pain was carried out. RESULTS: The mean age of the patients at the time of operation was 38.4 years (range, 11-78 years), and the mean follow-up period was 12.6 years (range, 3-36 years). Ten of the tumours were located in the median nerve distribution, whereas 4 in the ulnar nerve distribution. Four tumours were at the wrist level, 3 at the palm level, and 7 at the digital level (3 in the index finger). Only 5 patients were correctly diagnosed preoperatively. Neurological loss was not recovered in one patient in whom the tumour was resected with the digital nerve. No recurrence or muscle atrophy was noted at the latest follow-up. CONCLUSION: Meticulous dissection with magnification can achieve complete tumour removal without neurological loss or recurrence. Intracapsular tumour removal provides good results with a low complication rate. PMID- 16365491 TI - Periacetabular limb salvage for malignant bone tumours. AB - PURPOSE: To evaluate treatment outcomes in primary malignant periacetabular bone tumour removal and limb salvage with or without bone-graft reconstruction. METHODS: A total of 13 patients were treated for malignant periacetabular bone tumours at Siriraj Hospital, Bangkok, Thailand. The diagnoses were chondrosarcoma (n=8), Ewing's sarcoma (n=2), osteosarcoma (n=1), well-differentiated osteosarcoma (n=1), and malignant giant cell tumour (n=1). 11 patients did not undergo reconstruction following tumour resection; 2 patients received fibular bone grafts bridging the periacetabulum to the remaining sacrum. Adjuvant chemotherapy was administered for high-grade malignant tumours, and postoperative radiation therapy was performed on patients with a closed surgical margin. RESULTS: At a mean follow-up of 24.3 months (range, 8.9-43.9 months), 9 patients remained disease-free, 3 had died of the disease, and one was alive with disease. According to the Musculoskeletal Tumor Society classification system, the mean functional analysis at final follow-up was 68.7%. Patients who underwent internal hemipelvectomy experienced a subsequent leg-length discrepancy ranging from 3 to 10 cm. Four patients had complications (one each for deep wound infection, skin necrosis, seroma, and vascular spasms) and were successfully treated with multiple debridements and appropriate antibiotics. Three patients had local recurrences; one required a classic hemipelvectomy. CONCLUSION: Malignant periacetabular tumours are difficult to manage. Functional results of our patients with no reconstruction or with bone-graft bridging were fair. Patients undergoing internal hemipelvectomy may experience leg-length discrepancies, which can be balanced with shoe lifts. PMID- 16365492 TI - Midterm results of cemented Press Fit Condylar Sigma total knee arthroplasty system. AB - PURPOSE: To evaluate the midterm results of 50 patients who underwent total knee replacement using Press Fit Condylar (PFC) Sigma system. METHODS: We retrospectively reviewed 87 consecutive cases (50 patients with 37 bilateral cases) of PFC Sigma total knee replacement performed between January 1998 and December 1999. Patients were evaluated clinically and radiographically by an independent observer. The American Knee Society Score, Oxford Knee Score, and Knee Society radiographic assessment were used to rate knee function and to determine the satisfaction level of each patient. RESULTS: The mean age of the patients at the time of operation was 65 years (range, 41-85 years). The mean follow-up period was 5.4 years (range, 4.5-6.4 years). 44 patients (79 knees) were available for follow-up, 3 patients (3 knees) were lost to follow-up, and 3 patients (5 knees) died of unrelated causes. At the final follow-up, the mean Oxford Knee Score was 22. Using the American Knee Society Score, 88% of the knees were rated excellent, 4% good, 2% fair, and 6% poor. Five knees required revision surgery, the indications being infection in 4 knees and aseptic loosening in one knee. The survival rate of the implants was 94% at 6 years. CONCLUSION: The PFC Sigma total knee arthroplasty system has demonstrated good midterm results at our institution. PMID- 16365493 TI - Alkalinisation of local anaesthetics prescribed for pain relief after surgical decompression of carpal tunnel syndrome. AB - PURPOSE: To compare the duration of analgesia achieved following administration of buffered prilocaine versus plain prilocaine to patients undergoing surgical decompression of the median nerve. METHODS: 40 (32 female and 8 male; mean age, 50.5 years) patients who underwent surgical decompression of carpal tunnel syndrome were recruited. Patients were randomly allocated to 2 groups: the alkalinised group was given 10 ml of prilocaine hydrochloride 2% buffered with 1 ml of sodium bicarbonate 8.4%, whereas the non-alkalinised group received 10 ml of plain prilocaine hydrochloride 2% solution. Patients were asked to rate their comfort level at the operation site in the first 6 hours following surgery and after discharge from hospital using a visual analogue scale (VAS). The duration of analgesic effect was evaluated every 3 hours. Additional oral analgesia in the form of paracetamol 500 mg tablets was available to patients if required for break-through pain. RESULTS: Significantly lower VAS scores were reported by the alkalinised group during the first postoperative 12 hours. The change of VAS scores over time was significantly higher in the non-alkalinised group. The mean analgesic requirement for paracetamol tablets in the alkalinised and non alkalinised groups was 4 and 34, respectively. CONCLUSION: Buffered prilocaine provided a longer postoperative pain-free period for patients undergoing surgical decompression of the median nerve. It is easy, safe, and cost-effective. PMID- 16365494 TI - Comminuted refracture of the distal femur and condyles in patients with an intramedullary nail: a report of 5 cases. AB - Patients with in situ intramedullary nails sustaining a second distal femoral fracture following high-velocity trauma are infrequently reported in the literature. We report 5 such patients who sustained a second injury after 6 to 24 months, resulting in another fracture at the nail-tip level. These second fractures severely comminuted the femoral condyles. These patients required removal of the intramedullary nail, reconstruction of the comminuted femoral condyles, and stabilisation using an angled or buttress plate. Four of the 5 patients were followed up for a mean period of 36 months and were evaluated for knee status and outcome using Judet's criteria. Postoperative knee stiffness was common, and further interventions such as manipulation, hardware removal, arthrolysis, and quadricepsplasty were needed. One patient was lost to follow-up. The final results of the knees were good, with more than 100 degrees range of movement. It is important to adhere to established internal fixation protocols in the treatment of these complex nail tip fractures. PMID- 16365495 TI - Terrible triad of the elbow--role of the coronoid process: a case report. AB - Posterior dislocation of the elbow joint with fracture of both the radial head and coronoid process, the so-called 'terrible triad', is a complex and difficult to-treat injury. We report a case of a 34-year-old man with terrible triad of the elbow and associated injuries secondary to a fall. The patient was treated with radial head osteosynthesis, resection of the coronoid process fragments, and stabilisation of the ulnohumeral joint by Kirschner wire transfixation. The sequence of treatment steps is provided. The priority for treating multiple injuries should be established. The integrity of the coronoid process is essential for providing the elbow with maximum functionality. PMID- 16365496 TI - Iatrogenic bilateral pneumothorax arising from acupuncture: a case report. AB - Acupuncture is often regarded as innocuous. However, its complication can be serious and deadly if unattended. We report a case of iatrogenic bilateral pneumothorax after acupuncture therapy. Setting up a government regulatory body and using needles with safety design can prevent further inadvertent incidences from occurring. PMID- 16365497 TI - Bilateral anterior dislocation of the shoulders with proximal humeral fractures: a case report. AB - Bilateral simultaneous anterior dislocation of the shoulders with bilateral 3 part fracture of the proximal humeri is unusual. A 42-year-old man presented with pain and restriction of movement on both shoulders. He was injured by a heavy object falling over his back while he was leaning forward holding an overhead bar. His arms were abducted and externally rotated. The injury was not correctly diagnosed, and the patient was treated with repeated manipulations and splintage for 2 weeks. Radiological examination revealed bilateral anterior dislocation of the shoulders with displaced 3-part fractures of the proximal humeri involving the shaft, greater tuberosity, and head. The patient was treated with open reduction and internal fixation through a deltopectoral approach using multiple Kirschner wires. The shoulders were kept immobilised for 3 weeks until the removal of the wires. The patient was able to resume work 3 months after surgery. He had an excellent and comfortable range of motion in both shoulders at one-year follow-up. PMID- 16365498 TI - Missed diagnosis of triceps tendon rupture: a case report and review of literature. AB - Rupture of the triceps tendon is an uncommon injury, probably the rarest of all tendinous ruptures. We present a case of missed diagnosis of triceps tendon rupture. Such diagnosis should be considered when a patient presents with pain and swelling at the back of the elbow after a fall on an outstretched hand. PMID- 16365499 TI - Greater arc injury of the wrist with fractured lunate bone: a case report. AB - Carpal dislocation and fracture dislocation are uncommon and difficult to treat. Early diagnosis and treatment of such injuries are necessary to prevent progressive carpal instability and traumatic arthritis. Perilunate fracture dislocation is a combination of ligamentous and osseous injuries that involve the 'greater arc' of the perilunate. Despite being severe, these injuries often go unrecognised in the emergency department, leading to delayed diagnosis and treatment. We present a case of greater arc injury of the right wrist with fractures of the lunate and ulnar styloid without perilunate dislocation. This pattern of injury cannot be classified in the available literature on greater arc injury. PMID- 16365500 TI - Diabetic muscle infarction of the peroneus brevis: a case report. AB - Diabetic muscle infarction of the peroneus brevis is a rare complication of diabetes mellitus. It often presents with an acute leg pain without any obvious signs. Due to poor awareness of this condition, it is often misdiagnosed and results in anxiety of the patient and unnecessary investigations. It is ominous as it usually indicates vascular disease of the end organs. We report a case where the patient presented to several physicians for leg pain without any other clinical signs. The diagnosis was suspected on ultrasound and confirmed with biopsy. The symptoms improved dramatically after excision. Although most literature suggests conservative treatment by non-weight bearing and analgesia, the recovery period can last for more than 6 weeks, with a recurrence rate of 50%. We feel that there is a role for excision biopsy of the lesion for both diagnosis and treatment. In this case, symptoms improved remarkably and the patient was able to ambulate without pain 2 days after the operation without needing any analgesia. PMID- 16365501 TI - Osteoid osteoma treated with computed tomography-guided percutaneous radiofrequency ablation: a case series. AB - The nidi of osteoid osteoma are small and difficult to identify precisely; this can necessitate substantial resection of the surrounding normal bone. We applied percutaneous, image-guided radiofrequency ablation to treat 5 patients with osteoid osteoma. The precise location of each nidus was determined using a 3-mm thick spiral computed tomographic scan. The most appropriate needle pathway was selected to avoid major neural and vascular structures. A 14-gauge core needle was introduced into the nidus and then exchanged for a 17-gauge straight rigid electrode with a 1-cm exposed tip to heat the lesion to 90 degrees centigrade for 5 minutes. Four of the 5 patients were discharged on the day after surgery without any external supports, and were pain-free after a few days. The remaining patient had a pes equinus contracture induced by the penetration of the calf muscles through a posterior approach but was fully recovered 6 days later. The mean follow-up period was 20.6 months. No recurrences or late complications were observed in this series. This case series shows that the computed tomography guided percutaneous radiofrequency ablation is a simple, minimally invasive, and highly effective technique for the treatment of osteoid osteoma. PMID- 16365502 TI - Hydatid bone disease: a case report and review of the literature. AB - Hydatid disease may develop in almost any part of the body and can be identified with a combination of clinical history, imaging findings, and serologic results; however, the diagnosis of bone hydatidosis is primarily based on radiographic findings. Bone hydatid disease is often asymptomatic, and its diagnosis is usually made at an advanced stage when lesions have become extensive. We present a case of a 45-year-old woman who was admitted to the University Hospital, Stara Zagora, Bulgaria complaining of pain in her left tibia. Radiographs revealed an oval cyst with a diameter of 3.5 cm, located in the diaphyseal part of the tibia. The cyst was excised, and no recurrence was observed on follow-up. Functional outcome was excellent. PMID- 16365503 TI - Recurrent spinal meningioma: a case report with review of the literature. AB - Spinal meningioma generally responds favourably to surgical excision and has a low rate of recurrence. However, follow-up data on patients under 50 years of age are limited. We report a recurrence of intraspinal meningioma 18 years after the initial surgery in a 45-year-old woman. She presented with weakness in both lower limbs that had gradually progressed over the previous 8 months, and difficulty in walking for the last 2 months. She had no history of sensory loss or visceral involvement, but had undergone a major surgical intervention of the upper spine for a similar episode of weakness in both lower limbs 18 years earlier. She underwent a second surgery at the same site, and a greyish-white tumour was excised, which was histopathologically confirmed as recurrent psammomatous meningioma. The patient demonstrated complete neurological recovery in 8 months. PMID- 16365504 TI - What is the meaning of standard venous admixture formula results in septic patients? AB - The standard venous admixture formula is widely used in the bedside assessment of intrapulmonary shunt in intensive care units. The intrapulmonary shunt fraction calculated by the standard venous admixture formula is affected by the systemic oxygen extraction ratio and thus reflects both systemic and intrapulmonary shunts, especially in septic patients with decreased oxygen extraction ratios. The standard venous admixture formula may cause misestimation of the intrapulmonary shunt fraction, especially in septic patients. Inert gas rebreathing techniques and simultaneous measurement of cardiac output by thermodilution and oxygen consumption by indirect calorimetry may be useful in septic patients. PMID- 16365505 TI - Pulse oximetry in the oesophagus. AB - Pulse oximetry has been one of the most significant technological advances in clinical monitoring in the last two decades. Pulse oximetry is a non-invasive photometric technique that provides information about the arterial blood oxygen saturation (SpO(2)) and heart rate, and has widespread clinical applications. When peripheral perfusion is poor, as in states of hypovolaemia, hypothermia and vasoconstriction, oxygenation readings become unreliable or cease. The problem arises because conventional pulse oximetry sensors must be attached to the most peripheral parts of the body, such as finger, ear or toe, where pulsatile flow is most easily compromised. Since central blood flow may be preferentially preserved, this review explores a new alternative site, the oesophagus, for monitoring blood oxygen saturation by pulse oximetry. This review article presents the basic physics, technology and applications of pulse oximetry including photoplethysmography. The limitations of this technique are also discussed leading to the proposed development of the oesophageal pulse oximeter. In the majority, the report will be focused on the description of a new oesophageal photoplethysmographic/SpO(2) probe, which was developed to investigate the suitability of the oesophagus as an alternative monitoring site for the continuous measurement of SpO(2) in cases of poor peripheral circulation. The article concludes with a review of reported clinical investigations of the oesophageal pulse oximeter. PMID- 16365506 TI - A computer-based approach to assess the perception of composite odour intensity: a step towards automated olfactometry calibration. AB - The 2004 Nobel Prize in Physiology or Medicine laureates, Richard Axel and Linda Buck, have made smell a less enigmatic sense to study. In clinical routine, olfactory function is assessed using defined concentrations of a single defined substance, a setting which is uncommon in daily life. The present study was therefore conducted to evaluate the applicability of composite odours. Air was contaminated with different quantities of cyclohexanol, cyclohexanone and cyclohexane to generate 73 gas mixtures (one component: n = 21, two components: n = 40, three components: n = 12). The intensity of perception was estimated for each mixture by an average of 60.3 healthy individuals (4,403 assessments). An artificial neural network (ANN) was trained and validated using the contaminants' concentrations with the corresponding estimated intensities. The inter-rater variability was low, as 75.7% of the assessments did not exceed a difference beyond 0.5 from the corresponding median (considered correct predictions). The ANN correctly estimated 78.1% of the gas mixtures, and in terms of the regression task the ANN demonstrated a sufficient prediction performance (Pearson's correlation coefficient r = 0.883; R(2) = 0.757) and outperformed linear regression (r = 0.770; R(2) = 0.667). Evaluating extra ANNs for gas mixtures comprising one, two or three components, the predictive power did not decrease when complexity increased. The aforementioned results reflect nonlinearity in human perception. ANN technology helps simulate human perception of composite odour intensity which may be applicable to olfactometry calibration and systems biological mathematical modelling. The use of composite odours may represent real life problems more adequately than single substances. PMID- 16365507 TI - A longitudinal study of breath isoprene in healthy volunteers using selected ion flow tube mass spectrometry (SIFT-MS). AB - Thirty volunteers (19 males, 11 females) were recruited for a 6-month study of the volatile compounds in their exhaled breath using the selected ion flow tube mass spectrometry (SIFT-MS) analytical technique. Volunteers provided weekly breath samples between 8:45 am and 1 pm (before lunch), and the concentrations of several trace compounds were obtained. In this paper, we focus on the isoprene in alveolar breath, which was monitored by SIFT-MS using NO(+) precursor ions. The mean isoprene level for all samples was 118 parts per billion (ppb) with a standard deviation of 68 ppb and the range of values for breath samples given is 0-474 ppb. Variability in isoprene levels was similar in most volunteers. Isoprene levels increased immediately after moderate exercise, but returned to normal within 2-3 min for those few volunteers that were investigated. Cholesterol levels analysed for only three of the subjects were not obviously correlated with isoprene concentration in breath. Differences in isoprene levels were not directly correlated to gender, age or body mass index. PMID- 16365508 TI - The influence of flow rate on breathing circuit compliance and tidal volume delivered to patients in mechanical ventilation. AB - Assessment of the gas volume that actually reaches the airways during mechanical ventilation appears to be a difficult task because of the presence of the breathing circuit. Most ventilators measure tidal volume at ventilator level making the determination of circuit compliance a critical factor in estimating the actual tidal volume. Tubing compliance can be measured in several ways and its value, being strongly dependent on the compressibility of the gas, may significantly differ depending on the measurement procedure. This paper addresses the dependence of the circuit compliance on the flow rate, and a theoretical hypothesis on the pneumatic behaviour of the breathing circuit is formulated and experimentally validated, with several tests conducted in vitro on an infant analogue. The dependence of the compliance on the inspiratory flow is experimentally assessed, and differences of about 20% on the measured value in the common flow range utilized in infant ventilation have been found, with consequent estimation errors of the volume delivered. Experimental tests show that the correct value of the tidal volume actually delivered to the patient can be reliably estimated from measurements performed at the ventilator level if the circuit compliance is determined with the same flow rates that will be utilized in mechanical ventilation. PMID- 16365509 TI - Objective selection of signals for assessment of cerebral blood flow autoregulation in neonates. AB - A number of different system identification techniques have been proposed to assess dynamic cerebral autoregulation in critically ill patients. From these methods, the response to a standard stepwise change in blood pressure can be estimated. Responses lacking physiological consistency are a common occurrence and could be the consequence of particular system identification procedures or, alternatively, caused by measurements with a poor signal-to-noise ratio. A multi observer approach was adopted in this paper to classify cerebral blood flow velocity (CBFV) step responses to spontaneous changes in arterial blood pressure in a group of 43 neonates with a mean gestational age of 33.7 weeks (range 24-42 weeks) and a mean birthweight of 1,980 g (range 570-3,910 g). Three experienced observers independently analysed the estimated step responses in 191 recordings each lasting 100 s; for an autoregressive (ARX) model, 124 (65%) of the step responses were accepted by at least two of the three observers. Two other system identification methods, transfer function analysis and the moving average Wiener Laguerre model, gave 90 (45%) and 98 (51%) acceptable responses, respectively. Only 54 epochs (28%) were accepted with all three methods. With 88 (46%) responses rejected by at least two methods, it can be concluded that signal quality was the main reason for nonphysiological step responses. To avoid the need for subjective visual selection, an automatic procedure for classifying step responses was implemented leading to sensitivities and specificities in the range 85-90%, with respect to the agreement with subjective evaluations. Objective selection of CBFV step responses is thus feasible and could also be adapted for other physiological measurement techniques relying on system identification methods. PMID- 16365510 TI - Thermography and colour duplex ultrasound assessments of arterio-venous fistula function in renal patients. AB - Vascular and clinical assessments of arterio-venous fistula (AVF) function and access are important in patients undergoing or preparing to undergo renal dialysis. Objective assessment techniques include colour duplex ultrasound and more recently medical infrared thermography. Ideally, these should help assess problems relating to fistula failure or to vascular steal from the hand which can result from excessive fistula blood flow. The clinical value of thermography, as yet, has not been assessed for this patient group. The aims of this study were therefore to investigate the relationships between thermography skin temperature measurement and (a) quantitative ultrasound measurement of AVF blood flow, and (b) qualitative clinical assessment of vascular steal from the hands. Fifteen adult patients underwent thermal imaging of the upper limbs, colour duplex ultrasound to derive AVF blood flow from brachial artery blood flow measurements, and a clinical evaluation for vascular steal. Temperature measurements were extracted from the thermograms, including bilateral arm and hand (Fistula -- Non Fistula) differences, for comparison with derived AVF blood flow and steal grading. Derived AVF blood flow ranged from 30 to 1,950 ml min(-1), with a mean rate close to one litre per minute. Thermography detected the warmer superficial veins in proximity to the patent fistulas, with bilateral differences in fistula region skin temperature correlated with derived AVF blood flow (using maximum temperature measurements the correlation was +0.71 [p < 0.01]; and using mean temperature measurements the correlation was +0.56 [p < 0.05]). When thermography measurements were compared with the clinical assessment of steal the mean hand temperature differences separated steal from non-steal patients with an accuracy of greater than 90%. In summary, we have now demonstrated the potential clinical value of medical infrared thermography for assessing AVF function in renal patients. PMID- 16365511 TI - A novel multiple frequency stimulation method for steady state VEP based brain computer interfaces. AB - The objective is to increase the number of selections in brain computer interfaces (BCI) by recording and analyzing the steady state visual evoked potential response to dual stimulation. A BCI translates the VEP signals into user commands. The frequency band from which stimulation frequency can be selected is limited for SSVEP. This paper discusses a method to increase the number of commands by using a suitable combination of frequencies for stimulation. A biopotential amplifier based on the driven right leg circuit (DRL) is used to record 60 s epochs of the SSVEP (O(z)-A(1)) on 15 subjects using simultaneous overlapped stimulation (6, 7, 12, 13 and 14 Hzs and corresponding half frequencies). The power spectrum of each recording is obtained by frequency domain averaging of 400 ms SSVEPs and the spectral peaks were normalized. The spectral peaks of the combination frequencies of stimulation are predominant compared to individual stimulating frequencies. This method increases the number of selections by using a limited number of stimulating frequencies in BCI. For example, six selections are possible by generating only three frequencies. PMID- 16365512 TI - Effects of 50 Hz magnetic field exposure on human heart rate variability with passive tilting. AB - The question of whether power-frequency magnetic fields of strengths relevant to industrial exposure can affect heart rhythm remains controversial. Because the reported effects on heart rate (HR) are so small, procedures which can provoke changes in the sympathovagal balance in a controlled manner may have a greater capacity for identifying subtle field-related changes, if they do exist. We have investigated HR and heart rate variability (HRV) spectral indices in 20 volunteers subjected to a tilt from the supine position to 60 degrees , head up. The tilting procedure was carried out under two conditions, field (28 microT resultant, circularly polarized) and sham, in a balanced double-blind design. Subjects were instructed to breathe in time with an audible cue at 2.5 s intervals. Although the anticipated significant changes in HR and the high frequency (HF), low frequency (LF) and LF/HF ratio (log transformed) occur with tilting, there were no significant differences between corresponding measures with and without exposure to magnetic fields (tilt ln LF/HF ratio 0.94 +/- 0.19 and 0.95 +/- 0.20 for sham and field, respectively). There was also no evidence of a field-related trend in spectral alterations when the time following tilting was divided into three 256 s epochs. PMID- 16365513 TI - Detection of lactate threshold by including haemodynamic and oxygen extraction data. AB - To date, few attempts have been made to correlate cardiovascular variables to lactate threshold (L(T)). This study was designed to determine the relationship between the accumulation of blood lactate and several haemodynamic variables during exercise. Eight male volunteer cyclists performed an incremental test on an electromagnetically braked cycle-ergometer consisting of a 50 W linear increase in workload every 3 min up to exhaustion. Blood lactate was measured with a portable analyser during each exercise step. Oxygen consumption (VO(2)) and pulmonary ventilation were measured by means of a mass spectrometer while heart rate, stroke volume and cardiac output (CO) were assessed by impedance cardiography. The arterio-venous oxygen difference (A-V O(2) Diff) was obtained by dividing VO(2) by CO. By applying the D(max) mathematical method, L(T) and thresholds of ventilatory and haemodynamic parameters were calculated. The Bland and Altman statistics used to assess agreement between two methods of measurement were applied in order to evaluate the agreement between L(T) and thresholds derived from ventilatory and haemodynamic data. The main result was that most of the haemodynamic variables did not provide thresholds which could be used interchangeably with L(T). Only the threshold of A-V O(2) Diff showed mean values that were no different compared to L(T) together with limits of agreement that were not very wide between thresholds (below +/-25%). Hence of the haemodynamic parameters, A-V O(2) Diff appears to be the one most closely coupled with lactate accumulation and consequently it is also the most suitable for non-invasive calculation of the L(T). PMID- 16365514 TI - Neonatal cerebral cortical lesion abolishes the anxiolytic action of diazepam in adult rats: effects of location and extent of cortical lesion. AB - In our previous study, diazepam (DZP), a benzodiazepine receptor agonist, failed to suppress foot-shock-elicited ultrasonic vocalizations (USVs) in adult rats that had been neonatally lesioned in the neocortex. Because neonatal lesion of the neocortex did not influence the production of USVs, the presence of an anxiolytic mechanism of DZP is suggested apart from any anxiogenic mechanism in the brain. However, the previous study did not indicate any specific cortical regional lesions that impaired the normal development of the anxiolytic mechanism in the brain. The present study was undertaken in order to examine whether neonatal lesion of the neocortex, smaller and more localized than that in the previous study, abolishes the anxiolytic effect of DZP on foot-shock-elicited and air-puff-elicited USVs. A neonatal lesion about 2 mm diameter was made in the unilateral frontal cortex frontal to the hindlimb area or in the occipital cortex caudal to the hindlimb area. The attenuating effect of DZP on the USVs elicited by both aversive stimuli was found to be abolished only in the frontal cortex lesioned rats. This finding indicates that the frontal cortex is likely to be specifically involved in the normal development of the benzodiazepine-anxiolytic mechanism in the brain. PMID- 16365515 TI - Transgenic mice over-expressing dicarbonyl/L-xylulose reductase gene crossed with KK-Ay diabetic model mice: an animal model for the metabolism of renal carbonyl compounds. AB - Carbonyl compounds in the blood stream tend to accumulate in the kidney of diabetic or end stage renal failure subjects. Previously we isolated cDNA encoding dicarbonyl/L-xylulose reductase (DCXR) from a mouse kidney cDNA library. In the present study, transgenic (Tg) mice were generated to study the functional role of DCXR in the kidney. With a six-fold increase in the DCXR protein expression levels in the kidney, the homozygous Tg mice did not show any notable histological abnormalities. While the elevated DCXR expression was observed throughout the body, its renal distribution was similar to that of the endogenous DCXR protein, namely, the major expression site was the collecting tubules, along with moderate expression in other tubules and Bowman's capsule, but it was absent from the interstitial area and glomeruli. The Tg mice were crossed with KK-A(y) diabetic model mice to examine the role of DCXR in the progression of diabetic nephropathy. The resulting progeny, Tg/A(y), showed lighter body weight, lower levels of blood glucose, water uptake and creatinine clearance compared to their +/A(y) littermates. Although remarkable pathological differences were not observed at the microscopic level and in the renal accumulation of carboxymethyl lysine, the data imply that DCXR might function in the metabolism of glucose or carbonyl compounds, and play a protective role in a kidney which is under hyperglycemic pressure. The DCXR Tg mice and the Tg x KK-A(y) hybrid mice, therefore, serve as specific models for carbonyl metabolism in the kidney with diabetic background. PMID- 16365516 TI - Spontaneous ophthalmic diseases in 586 New Zealand white rabbits. AB - The purpose of this study was to investigate spontaneous eye disease in New Zealand White (NZW) rabbits, which are commonly used for toxicity tests, and to provide reference materials for pharmaceutical companies and research centers. A total of 586 NZW rabbits were randomly chosen without sex preference and were examined using ocular equipment, including a direct ophthalmoscope, an indirect ophthalmoscope, a slit-lamp biomicroscope, a focal illuminator, and a fundus camera. This study showed that the incidence rate of temporary cataracts, regarded as a change within normal variation, was 0.5% in the NZW rabbits. Regarding abnormal ophthalmic disease, blepharitis was the most commonly observed ocular disease. Other findings included cataract, conjunctivitis, choroidal hypoplasia, keratitis, corneal scarring, eyelid laceration, posterior synechiae, uveitis, dacryocystitis, and persistent pupillary membrane. In total, the incidence rate of ophthalmic diseases was 9.6%. Based on sex and age distributions, females had more ocular diseases than males, and rabbits were less susceptible to eye diseases as they got older. In this study, photographs were taken to document findings, such as normal fundus, normal variations, ophthalmic disease, and histopathologic examination. PMID- 16365517 TI - Inhibitory effects of trientine, a copper-chelating agent, on induction of DNA strand breaks in kidney cells of Long-Evans Cinnamon (LEC) rats. AB - The effects of treatment with trientine, a specific copper-chelating agent, on the accumulation of copper and induction of DNA strand breaks were investigated in Long-Evans Cinnamon (LEC) rats, an animal model for human Wilson's disease. Copper accumulated in the kidneys of LEC rats in an age-dependent manner from 12 to 18 weeks of age. When LEC rats were treated with trientine from 10 weeks of age, renal copper contents did not increase and were maintained at the same levels as those in 4-week-old LEC rats. Estimation of the amounts of DNA single strand breaks (SSBs) by comet assay showed that SSBs of DNA were induced in a substantial population of LEC rat renal cortex cells around 12 weeks of age and that the amounts of SSBs increased in an age-dependent manner from 12 to 18 weeks of age. When LEC rats were treated with trientine from 10 weeks of age, the observed number of cells with DNA damage decreased, suggesting that induction of SSBs of DNA was inhibited and/or SSBs were repaired during the period of treatment with trientine. The results show that SSBs of DNA in LEC rat kidney cells are induced prior to occurrence of clinical signs of hepatic injury and that treatment of LEC rats with trientine decreases the number of DNA strand breaks. PMID- 16365518 TI - Discrimination of recent ischemic myocardial changes in WHHLMI rabbits from the findings of postmortem degeneration. AB - To distinguish recent ischemic myocardial changes in myocardial infarction-prone Watanabe heritable hyperlipidemic (WHHLMI) rabbits from general postmortem myocardial degeneration, we examined hearts of WHHLMI rabbits after sudden death and postmortem hearts of non-atherogenic rabbits. Hearts of 8 WHHLMI rabbits were excised within 30 min of sudden death and hearts of 27 non-atherosclerotic rabbits were excised at designated periods after sacrifice. A large number of myocardial cells from WHHLMI rabbits exhibited features characteristic of ischemia (intercellular gap, intracellular edema, eosinophilia, disappearance of myocardial cells, indistinct nuclei, wavy myocardial fibers) simultaneously at regions close to proximal occluded coronary arteries. Although postmortem hearts of non-atherosclerotic rabbits exhibited similar characteristics, several features characteristic of autolyzed myocytes were also randomly observed in the left ventricle wall. Each feature was detected independently in myocardial cells or regions of the ventricle wall. In conclusion, we found several unique characteristics associated with myocardial infarction which enable discrimination between recent ischemic myocardial changes and myocardial degeneration following death. PMID- 16365519 TI - Safe and efficient collection of cytokine-mobilized peripheral blood cells from cynomolgus monkeys (Macaca fascicularis) with human newborn-equivalent body weights. AB - Hematopoietic stem cells in bone marrow can be mobilized into peripheral blood by cytokine administration. Cytokine-mobilized peripheral blood stem cells are of great use in clinical applications. We previously established a modified procedure for the collection of cytokine-mobilized peripheral blood cells from rhesus monkeys (Macaca mulata) using a commercially available apparatus originally developed for human subjects. In this study, we examined the efficacy and safety of this method with even smaller macaques, cynomolgus monkeys (Macaca fascicularis), which are equivalent to human newborns in body weight (mean = 3.3 kg). Using the manufacturer's unmodified protocol (n=6), one monkey died of cardiac failure and three developed severe anemia. In contrast, using our modified procedure (n=6), no such complication was observed in any animal. In addition, the harvested nuclear cell, mononuclear cell and CD34(+) cell counts were significantly higher with the modified method. The modified method should allow safe and efficient collection of cytokine-mobilized peripheral blood cells from non-human primates as small as human newborns in a non-invasive manner. PMID- 16365520 TI - Development of dry skin in the NOA mouse under individual housing conditions: a potentially useful animal model for evaluating moisturizing effects. AB - In a previous study, we reported the development of grossly observable dry skin in all of the Naruto Research Institute Otsuka Atrichia (NOA) mice that were housed individually. In the present study, dermal physiological function tests were conducted and the usefulness of this dry skin model for evaluating the efficacy of topical moisturizers was assessed. As a result, we have confirmed a marked reduction in the water content of the stratum corneum in these animals. Therefore, the development of dry skin in the NOA mouse strain under individual housing conditions may be expected to serve as a useful animal model for evaluating topical moisturizers. Specifically, the water content of the stratum corneum was restored in proportion to the oil content of the ointment base used to treat the animals, and the moisturizing effects of urea were confirmed in animals treated with urea-containing ointment. In addition, when the animals that had been housed individually were returned to group housing conditions, the water content of the stratum corneum was restored, with a corresponding improvement in dry skin. This finding suggests that socio-psychological factors are involved in the etiology of dry skin in individually housed NOA mice. PMID- 16365521 TI - Development of intestinal microbiota in mice and its possible interaction with the evolution of luminal IgA in the intestine. AB - The development of the intestinal microbiota and the evolution of the fecal IgA in mice were analyzed from 18 to 40 days old by PCR temperature gradient gel electrophoresis (TGGE) and ELISA, respectively. There were two events for the diversification of the intestinal microbiota from suckling to maturity. The first change occurred between days 21 and 22 after birth, when the diversity of the intestinal microbiota showed a remarkable increase at this time. The second change occurred from days 27 to 30 after birth, and the increase in the diversity of the intestinal microbiota ceased. The amount of fecal IgA decreased from days 18 to 20, remained low until day 22, on day 23, it recovered and then continued to increase. This study suggests that there are possible interactions between the development of intestinal microbiota and the evolution of intestinal secretion of IgA in mice, the same as in rats, although the second change in mice intestinal microbiota occurred a few days later than in rats. The decline in maternal IgA supply as the suckling period proceeded presumably allowed the bacterial colonization. As a consequence of this increase in bacterial colonization, the secretion of the self-SIgA was accelerated in the pups. PMID- 16365522 TI - Strain-dependent differences in locomotor activity after local brain irradiation with 30 GyE of carbon ions. AB - This study investigated strain differences in brain damage among male A/J, C57BL/6JNrs and C3H/HeNrs mice after local brain irradiation. Whole brains were irradiated with a single dose of 30 GyE carbon ion beams and then locomotor activity was determined as body heat of each animal. The daily locomotor activities of untreated mice differed among strains. Non-irradiated C57BL/6JNrs mice were more active than A/J mice. This variance became more obvious immediately after irradiation, when the activity of A/J and C3H/HeNrs mice diminished, whereas that of C57BL/6JNrs mice increased at the beginning of the active phase and remained elevated for three days after irradiation. The altered activities of all three strains of irradiated mice gradually recovered to normal within three to four days. PMID- 16365523 TI - Immunolocalization of steroidogenic enzymes in the fetal, neonatal and adult testis of the Shiba goat. AB - The localizations of steroidogenic enzymes (P450scc, 3 beta HSD, P450c17 and P450arom) in testes of Shiba goats were investigated by immunohistochemistry. P450scc, 3 beta HSD, P450c17 and P450arom were detected in all Leydig cells of adults. P450scc and P450c17 were observed in most Leydig cells in the fetus (90 days) and neonate (15 days). 3 beta HSD and P450arom were found in some Leydig cells of the fetus with weak immunostaining but the numbers of immunopositive Leydig cells and intense immunostaining were increased in Leydig cells of the neonate. These results suggest that Shiba goat testes have the ability to synthesize progestin, androgen and estrogen in the fetus, neonate and adult, and synthesis of these steroid hormones showed an age-related rise. PMID- 16365524 TI - Rescue from dwarfism by thyroid function compensation in rdw rats. AB - The rdw rat was initially reported as having hereditary dwarfism caused by pituitary dysfunction. Subsequent studies on the rdw rat, however, have demonstrated that the primary cause of rdw dwarfism is present in the thyroid gland but not in the pituitary gland. The primary cause of rdw rat disorders is a missense mutation of the thyroglobulin (Tg) gene by a one-point mutation. In the present study, we attempted to rescue the dwarfism of the rdw rats using a diet supplemented with thyroid powder (T-powder) and a thyroid graft (T-graft). The infants of the rdw rat were successfully raised to a mature stage body weight, accompanied by elevation of serum growth hormone (GH) and prolactin (PRL), by the T-powder. Furthermore, the T-graft successfully increased the body weight with fertility. The serum GH and PRL levels in the T-graft rdw rat significantly increased. The serum thyroid-stimulating hormone (TSH) levels in the T-graft rdw rat were significantly decreased but were significantly higher than those in the control rat. The GH and PRL mRNA expression in the rdw rat with the T-graft was virtually the same as that of the control, but the TSH beta mRNA differed from that of the control rats. Thus, the dwarfism in the rdw rat is rescued by thyroid function compensation, such as that afforded by T-powder and T-graft. PMID- 16365525 TI - WBN/Kob-Ht rats spontaneously develop dermatitis under conventional conditions: another possible model for atopic dermatitis. AB - WBN/Kob-Ht rats (Ht-rats) raised under conventional conditions spontaneously developed dermatitis. In this study, we carried out histopathological analysis to elucidate the pathological features of the dermatitis in Ht-rats. We then tried to detect Staphylococcus species recovered from the skin lesions of Ht-rats. We also measured the serum levels of total IgE, IL-4 and IFN-gamma in these rats. The histopathological data indicated that inflammatory cells had infiltrated the skin lesions. Staphylococcus aureus was recovered from the skin lesions, and the serum levels of total IgE and IL-4 were elevated in Ht-rats with dermatitis. These results suggest that dermatitis in Ht-rats is similar to that in the DS-Nh mice, which has recently been proposed as an animal model for human atopic dermatitis. PMID- 16365526 TI - stx2vha is the dominant genotype of Shiga toxin-producing Escherichia coli O157:H7 isolated from patients and domestic animals in three regions of China. AB - Shiga toxin-producing Escherichia coli(STEC) O157: H7 strains were isolated from domestic animals and patients from Xuzhou City, Jiangsu Province, China and the bordering Anhui and Henan Provinces and were examined for the stx genotype. Of 390 strains, 277 were identified as genotype stx2vha ; 41, stx2 ; 51, stx2-stx1 ; 1, stx2-stx2vha-stx1 ; 5, stx2-stx2vha ; and 15 were un-typeable. Of the 277 stx2vha-bearing isolates, 116 were isolated from goats; 42, cattle; 38, hens, and 35 from pigs. The study shows stx2vha is the dominant genotype and goats are an important reservoir. PMID- 16365527 TI - Legionella pneumophila serogroup 1 isolates from cooling towers in Japan form a distinct genetic cluster. AB - Thirty-one epidemiologically unrelated Legionella pneumophila serogroup 1 isolates (10 from cooling towers, 10 from public spas and/or hot spring baths, and 11 from patients) were analyzed by pulsed-field gel electrophoresis (PFGE) and sequence-based typing (SBT) using 6 loci, flaA, pilE, asd, mip, mompS, and proA. The results of PFGE and SBT analysis indicated that all 10 isolates from cooling towers clustered into a unique type, which was distinct from strains of other environmental sources. PMID- 16365528 TI - Evaluation of the Lon-deficient Salmonella strain as an oral vaccine candidate. AB - We evaluated the efficacy of CS2022 (the Lon protease-deficient mutant strain of Salmonella enterica serovar Typhimurium) as a candidate live oral vaccine strain against subsequent oral challenge with a virulent strain administered to BALB/c and C57BL/6 mice. CS2022 persistently resided in the spleen, mesenteric lymph nodes, Peyer's patches, and cecum of both strains of mice after a single oral inoculation with 1 x 10(8) colony-forming units. Finally, CS2022 almost disappeared from each tissue sample by week 12 in BALB/c mice, whereas CS2022 still resided in each tissue type at week 12 after inoculation of C57BL/6 mice. A significant increase in the serovar Typhimurium lipopolysaccharide-specific secretory immunoglobulin A (s-IgA), as measured for one of the mucosal immune responses, was detected in bile and intestinal samples of both strains of immunized mice at week 4 after immunization. In addition, the expression of gamma interferon mRNA in the spleens of both strains of immunized mice, especially those of C57BL/6 mice, was significantly increased at week 4 after immunization and was boosted during the following 5 days after the challenge was administered to the mice. Furthermore, peritoneal macrophages isolated from immunized mice at week 4 after immunization exhibited an increase in intracellular killing activity against both virulent and avirulent Salmonella. The present results suggested that salmonellae-specific s-IgA on the mucosal surfaces induced by immunization with CS2022 generally prevented mice from succumbing to an oral challenge with a virulent strain. Simultaneously, CS2022 promoted the protective immunity associated with macrophages in both strains of mice. PMID- 16365529 TI - Inhibitory effect of RNAi on Japanese encephalitis virus replication in vitro and in vivo. AB - Flaviviruses include many insect-mediated small viruses and still cause serious problems in the world. In humans, JEV can cause acute meningioencephalomyelitis, resulting in fatality rates of 5 to 40%. RNA-interference (RNAi) as an antiviral mechanism was originally discovered in plants and then found in the specific suppression of gene expression of other organisms such as Caenorhabditis elegans, Drosophila and vertebrates. As JEV is an RNA virus, RNAi could be a reasonable approach for therapeutic purposes to use against Japanese encephalitis. In this study, we examined the effect of RNAi on JEV replication. Viral reproduction in Vero cells was decreased to 7.2% and 39.0% of control by the transfection of small interference RNAs, JCR and JN3R at 250 n M, respectively. Under the transfection of 5 microg/ml pJRi which produces stem-loop RNAi, viral reproduction was decreased to about 10% of control. Western blot analysis indicated that RNAi inhibited the translation level. We used pJRi in the animal experiment. After the inoculation of viruses at 5 x 10(3) PFU, pJRi at 1.0 and 5.0 microg/g was injected into mice i.p. JEV-infected control mice (n=5) died within 15 days. pJRi (1.0 or 5.0 microg/g)-medicated mice survived 40 or 80% at 15 days. The data clearly indicate that pJRi has highly potent inhibitory activity against JEV replication in vivo. The results in vivo and in vitro provide evidence that JEV replication was efficiently inhibited by RNAi and RNAis could be used as an antiviral drug against JEV infection. PMID- 16365530 TI - Evaluation of the biofilm-forming ability and genetic typing for clinical isolates of Pseudomonas aeruginosa by enterobacterial repetitive intergenic consensus-based PCR. AB - Biofilm formation is an important phenotype associated with chronic Pseudomonas aeruginosa infections. In the present study, a total of 48 P. aeruginosa strains isolated from clinical specimens were examined for their biofilm-forming ability using a microtiter plate method. The different biofilm-forming abilities were demonstrated among the strains; however, most strains formed a larger biofilm than strain PAO1, a reference strain. The genetic typing was also carried out by enterobacterial repetitive intergenic consensus-based polymerase chain reaction. Although they were divided into five groups (A to E), most of the strains showing the higher biofilm-forming ability were found to be in groups D and E, suggesting a significant relationship between the biofilm-forming ability and the genetic group. PMID- 16365531 TI - Evaluation of the ESPLINE INFLUENZA A&B-N Kit for the diagnosis of avian and swine influenza. AB - The ESPLINE INFLUENZA A&B-N kit was evaluated for its applicability to the rapid diagnosis of influenza in chickens and pigs. The kit specifically detected viral antigens in tracheal swabs and tissue homogenates of the trachea, liver, spleen, and colon of chickens inoculated with a highly pathogenic avian influenza virus strain, A/chicken/Yamaguchi/7/04 (H5N1), at 48 hr post-inoculation (p.i.) as well as in the tracheal and cloacal swabs and tissue homogenates of dead chickens. For those infected with a low pathogenic strain, A/chicken/aq-Y-55/01 (H9N2), antigens were detected only in the samples from tracheal swabs and organs 1-4 days p.i. The kit also detected viral antigens in the nasal swabs of miniature pigs infected with swine and avian influenza viruses. The kit was found to be sensitive and specific enough for the rapid diagnosis of infections of influenza A virus in chickens and pigs. PMID- 16365532 TI - Increased expression of c-myc is associated with thymoma in rats infected with murine leukemia virus A8. AB - Infection of rats with Friend murine leukemia virus (Fr-MLV) clone A8 causes thymoma in all the animals within 7 weeks. The rapid induction of thymoma is associated with a unique enhancer structure in the U3 region of the A8-LTR. Our Southern blot analyses showed that the thymomas were oligo clonal. The A8-induced thymomas showed 3-to 11-fold overexpression of c-myc mRNA. These results suggest that provirus insertion into particular positions of the host genome is correlated with tumorigenesis after A8 infection and that up-regulation of c-myc plays an important role in the induction of thymoma. PMID- 16365533 TI - Analysis of the distribution of neuropathogenic retroviral antigens following PVC211 or A8-V infection. AB - A8-V and PVC211 are neuropathogenic strains of the Friend murine leukemia virus (Fr-MLV) that cause spongiosis in the rat brain after infection at birth. PVC211 exhibited stronger neuropathogenicity than A8-V, and induced more severe neurological symptoms such as hind-leg paralysis. These symptoms correlated with the neuropathological spread and intensity, which were more severe in the spinal cord of rats infected with PVC211 than in those infected with A8-V, without exhibiting neuropathological differences in other areas of the CNS. Interestingly, virus titers recovered from infected spinal cords were similar in PVC211 and A8-V infected animals. However, in the spinal cord infected with PVC211, glial cells attained higher immunohistochemical expression scores for the viral surface antigen, gp70 (Env) than in the A8-V infected spinal cord, although expression levels of viral antigens in blood vessel walls were similar in A8-V and PVC211 infections. Furthermore, many of those glial cells which carried viral antigens were found, by double immunostaining, to be microglia. The results suggested that the spread of viral antigen positive microglia plays an important role in forming the different neuro-pathogenicity observed in A8-V and PVC211 infections. PMID- 16365534 TI - Protective efficacy of nonpathogenic nef-deleted SHIV vaccination combined with recombinant IFN-gamma administration against a pathogenic SHIV challenge in rhesus monkeys. AB - We previously reported that a nef-deleted SHIV (SHIV-NI) is nonpathogenic and gave macaques protection from challenge infection with pathogenic SHIV-C2/1. To investigate whether IFN-gamma augments the immune response induced by this vaccination, we examined the antiviral and adjuvant effect of recombinant human IFN-gamma (rIFN-gamma) in vaccinated and unvaccinated monkeys. Nine monkeys were vaccinated with nef-deleted nonpathogenic SHIV-NI. Four of them were administered with rIFN-gamma and the other five monkeys were administered with placebo. After the challenge with pathogenic SHIV-C2/1, CD4(+) T-cell counts were maintained similarly in monkeys of both groups, while those of the unvaccinated monkeys decreased dramatically at 2 weeks after challenge. However, the peaks of plasma viral load were reduced to 100-fold in SHIV-NI vaccinated monkeys combined with rIFN-gamma compared with those in SHIV-NI vaccinated monkeys without rIFN-gamma. The peaks of plasma viral load were inversely correlated with the number of SIV Gag-specific IFN-gamma-producing cells. In SHIV-NI-vaccinated monkeys with rIFN gamma, the number of SIV Gag-specific IFN-gamma-producing cells of PBMCs increased 2-fold compared with those in SHIV-NI-vaccinated monkeys without rIFN gamma, and the NK activity and MIP-1alpha production of PBMCs were also enhanced. Thus, vaccination of SHIV-NI in combination with rIFN-gamma was more effective in modulating the antiviral immune system into a Th1 type response than SHIV-NI vaccination alone. These results suggest that IFN-gamma augmented the anti-viral effect by enhancing innate immunity and shifting the immune response to Th1. PMID- 16365535 TI - In New Jersey, an effort to reduce pressure ulcers. PMID- 16365541 TI - Atypical wounds: recognizing and treating the uncommon. PMID- 16365542 TI - Do patients with pressure ulcers benefit from oral zinc supplementation? PMID- 16365543 TI - 3 cases of aberrant wound healing in acute myelogenous leukemia. PMID- 16365544 TI - Concerns voiced over competitive bidding of support surfaces. PMID- 16365545 TI - The PUSH tool: a survey to determine its perceived usefulness. AB - OBJECTIVE: To determine the perceived usefulness of the Pressure Ulcer Scale for Healing (PUSH). PARTICIPANTS: A convenience sample identified through the National Pressure Ulcer Advisory Panel Web site as users or registered users of the PUSH tool. MAIN OUTCOME MEASURE: A survey instrument was developed to capture experience, ease of use, and perceived utility and weakness of the PUSH tool. RESULTS: Of 103 respondents, most (79) agreed or strongly agreed that PUSH required an appropriate amount of time to complete. It was also found to be reliable and easy to use and teach to others. Respondents were not as positive regarding usefulness, with 75% indicating that increased PUSH scores prompt patient and treatment reassessment. Respondents agreed or strongly agreed that improvement is possible in the size subscale (59%), the tissue type subscale (49%), and the exudate amount subscale (32%). Most commonly indicated for improvement was the addition of wound depth information. CONCLUSION: Respondents generally found PUSH easy to use and helpful in pressure ulcer management. Specific areas of improvement were also identified. PMID- 16365546 TI - Wound debridement with 25 kHz ultrasound. PMID- 16365551 TI - Imaging techniques for uveal melanoma. PMID- 16365547 TI - The role of therapeutic angiogenesis in tissue repair and regeneration. AB - PURPOSE: To provide the physician and registered professional nurse with an understanding of angiogenesis and an overview of therapeutic angiogenesis modalities used to manage wounds and other tissue repair situations. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in learning more about angiogenesis and therapeutic angiogenesis modalities to manage wounds and other tissue repair situations. OBJECTIVES: After reading the article and taking the test, the participant should be able to: PMID- 16365552 TI - Laser for choroidal melanoma. PMID- 16365553 TI - Future laser approaches in melanoma treatment. PMID- 16365554 TI - Proton teletherapy of uveal melanoma. PMID- 16365555 TI - Radioactive plaque therapy. PMID- 16365556 TI - Newer radiation modalities for choroidal tumors. PMID- 16365557 TI - The role of eyewall resection in uveal melanoma management. PMID- 16365558 TI - Neoadjuvant proton beam radiotherapy combined with subsequent endoresection of choroidal melanomas. PMID- 16365559 TI - Surgical techniques in enucleation: the role of various types of implants and the efficacy of pegged and nonpegged approaches. PMID- 16365560 TI - Metastatic uveal melanoma. PMID- 16365561 TI - Metastatic uveal melanoma therapy: current options. PMID- 16365562 TI - Molecular pathobiology of uveal melanoma. PMID- 16365564 TI - Endovaginal magnetic resonance imaging of the female urethra. AB - Magnetic resonance imaging (MRI) is playing an important role in the clinical evaluation of women presenting with urethral symptoms. Voiding cystourethrography, direct urethrography, and pelvic sonography provide limited information on abnormalities that are in continuity with the urethra. On the other hand, urethra and periurethral tissues can be noninvasively evaluated by high-resolution endocavitary MRI. Because of its multiplanar capability and high tissue contrast, endovaginal MRI is an extremely reliable diagnostic test in the evaluation of urethral abnormalities. In this article, the utility of endovaginal MRI in the detection and characterization of a wide spectrum of urethral pathologic conditions, such as congenital anomalies, diverticula, urethritis, and benign and malignant neoplasms, is discussed. PMID- 16365565 TI - Localization of prostate cancer using 3T MRI: comparison of T2-weighted and dynamic contrast-enhanced imaging. AB - OBJECTIVE: To compare dynamic contrast-enhanced imaging and T2-weighted imaging using a 3T MR unit for the localization of prostate cancer. METHODS: Twenty consecutive patients with biopsy-proven prostate cancer underwent both T2 weighted imaging and dynamic contrast-enhanced imaging. At T2-weighted imaging and dynamic contrast-enhanced imaging, the presence or absence of prostate cancer confined within the prostate without extracapsular or adjacent organ invasion was evaluated in the peripheral zones of base, mid-gland, and apex on each side. Final decisions on prostate cancer localization were made by consensus between two radiologists. Degrees of depiction of tumor borders were graded as poor, fair, or excellent. RESULTS: Prostate cancer was pathologically detected in 64 (53%) of 120 peripheral zone areas. The sensitivity, specificity, and accuracy for prostate cancer detection were 55%, 88% and 70% for T2-weighted imaging and 73%, 77%, and 75% for dynamic contrast-enhanced imaging, respectively. Three cancer areas were detected only by T2-weighted imaging, 15 only by dynamic contrast-enhanced imaging, and 34 by both T2-weighted imaging and dynamic contrast-enhanced imaging. A fair or excellent degree at depicting tumor border was achieved in 67% by T2-weighted imaging and in 90% by dynamic contrast enhanced imaging (P<0.05). CONCLUSIONS: Dynamic contrast-enhanced imaging at 3T MRI is superior to T2-weighted imaging for the detection and depiction of prostate cancer and thus is likely to be more useful for preoperative staging. PMID- 16365566 TI - Magnetic resonance imaging findings of small round cell tumors of the ovary: a report of 5 cases with literature review. AB - OBJECTIVE: To study the magnetic resonance imaging characteristics of the ovarian involvement of small round cell tumors. METHODS: Magnetic resonance imaging findings were retrospectively reviewed in 5 patients seen at this institution and in 7 patients reported in the literature with ovarian small round cell tumors. Laterality, signal intensity, existence of hemorrhage, calcification, necrosis, septa, cerebroid appearance, and peripheral cysts were evaluated. RESULTS: Seven patients had bilateral disease, and the others had unilateral disease. The signal intensity was low on T1-weighted imaging (T1WI) in all cases. Signal on T2 weighted imaging (T2WI) and the degree of contrast enhancement varied. No case showed hemorrhage or calcification. Only 1 case demonstrated necrosis. Septa were observed in 3 cases, and a cerebroid appearance was observed in 5 cases. Six patients of reproductive age had multiple small cysts at the periphery of the masses. CONCLUSION: Despite the limited number of cases, peripheral small cysts in the large lobulated solid mass seemed to be a characteristic of the ovarian small round cell tumors in patients of reproductive age. PMID- 16365567 TI - Computed tomography features of an intraductal polypoid mass: differentiation between hepatocellular carcinoma with bile duct tumor invasion and intraductal papillary cholangiocarcinoma. AB - OBJECTIVE: To determine the computed tomography (CT) features capable of differentiating hepatocellular carcinoma (HCC) with bile duct tumor invasion (BDTI) from intraductal cholangiocarcinoma (IDCC). METHODS: Multiphasic CT images of 14 patients with HCC with BDTI and 18 patients with IDCC were retrospectively reviewed. Analysis of the CT findings included the size, location, and margin of the intraductal mass; enhancement pattern of intraductal lesions; degree of ductal dilatation; presence of downstream ductal dilatation and ductal wall thickening; presence of a parenchymal mass and its size; continuity of the parenchymal mass with the intraductal mass; and liver cirrhosis. Objective evaluation of the enhancement patterns of intraductal tumors was done by measuring the CT attenuation coefficients of the tumors and the uninvolved hepatic parenchyma in each phase. Among these findings, statistically significant variables were then determined using the Fisher's exact test or Mann-Whitney U test. RESULTS: Significant variables that helped to differentiate HCC with BDTI from IDCC included the presence of a parenchymal mass, liver cirrhosis, and a hyperattenuating intraductal tumor on the hepatic arterial phase (HAP). On unenhanced images, the tumor-to-liver contrast of IDCC (16.7+/-8.1) was greater than that of HCC with BDTI (6.4+/-10.4), but on the HAP, that of HCC with BDTI (26.5+/-28.2) was greater than that of IDCC (5.9+/-18.7) (P<0.05). In addition, there was a significant difference in the enhancement ratio of the intraductal tumors on the portal venous phase between the 2 conditions (P=0.003). CONCLUSIONS: Several objective and subjective multiphasic CT findings may help to differentiate HCC with BDTI from IDCC. PMID- 16365568 TI - Does the CT whirl sign really predict small bowel volvulus?: Experience in an oncologic population. AB - The objective of this study was to determine the sensitivity of the CT "whirl sign" for the diagnosis of small bowel volvulus in patients who present with a clinical suspicion of intestinal obstruction. Between January 2002 and September 2004, 1,493 CT scans performed in 1,213 patients suspected of having small bowel obstruction were retrospectively reviewed by one attending radiologist with gastrointestinal subspecialization and one senior radiology resident. Multislice helical CT scans were performed after oral and intravenous contrast administration. All CT scans showing a combined vessel and bowel whirl appearance were identified. Other features recorded included the number of degrees of whirl rotation, direction of rotation, presence of bowel obstruction, and signs of ischemia. Diagnoses were determined at either surgery or clinicoradiographic follow-up. Surgical follow-up was available in 174 of the 1,213 patients. There were 460 males and 753 females ranging in age from 1 to 95 years (mean 59 years). A whirl sign was found in 33 of the 1,493 CT scans by reader 1 and in 13 of the 1,493 CT scans by reader 2. In 11 patients, surgery revealed small bowel volvulus (0.9%). Reader 1 detected 7 of the 11 volvuli (sensitivity 64%, specificity 98%, positive predictive value 21%, negative predictive value 99.7%). Reader 2 detected 3 of the 11 volvuli (sensitivity 27%, specificity 99%, positive predictive value 23%, negative predictive value 99.5%). The CT scans of the four remaining patients with volvulus not initially recognized by either reader were re-reviewed and were felt to contain whirl signs. Most patients with small bowel volvulus can be identified on CT through detection of a whirl sign. However, most whirl signs detected on CT will not prove to be indicative of small bowel volvulus. PMID- 16365569 TI - Multidetector CT urography: comparison of two different scanning protocols for improved visualization of the urinary tract. AB - The goal of this study was to evaluate different CT scanning protocols on the depiction of the urinary tract by multidetector CT. The authors retrospectively reviewed 55 consecutive patients who underwent CT scanograms. Two groups of patients were included: renal donors (n=29) and hematuria patients (n=26). For the renal donor protocol, 120 mL of iodinated contrast was injected and a CT scanogram was obtained after a 5-minute delay. For the hematuria CT urography protocol, 100 mL of contrast was followed by a 250 mL normal saline drip and CT scanograms acquired after an 8-minute delay. Urinary tracts from both imaging protocols were then divided into four segments and evaluated by consensus reading of two experienced radiologists rated on a scale of 0 to 2. Complete visualization of the renal pelvis and the proximal, middle, and distal ureter for the renal donor protocol was noted to be 86%, 57%, 45%, and 52% and for that of the CT urography protocol to be 75%, 65%, 40%, and 44%, respectively. Comparing scanograms of the renal donor protocol and the CT urography protocol, there was no statistically significant difference in the depiction of renal pelvis or the proximal, middle or distal ureteral region (P=0.1625, 0.3226, 0.8636, and 0.6145, respectively). The study demonstrates that there is no significant difference between the CT urography protocol and the renal donor protocol in the depiction of the urinary tract. PMID- 16365570 TI - Computed tomography features of nonalcoholic steatohepatitis with histopathologic correlation. AB - OBJECTIVE: This study was conducted to describe the computed tomography (CT) features of nonalcoholic steatohepatitis (NASH) and to evaluate if the CT features could be used to diagnose and stage NASH. METHODS: From 1994 until 2004, pathology records revealed 68 patients with NASH. Of these, 12 patients underwent CT scans before (n=6), on the same day as (n=3), or after (n=3) a liver biopsy. Using the same database, 9 patients with steatosis alone evaluated with a CT scan before (n=2), on the same day as (n=3), or after (n=4) the liver biopsy were selected as a control group. Two radiologists measured liver attenuation (compared with spleen) and assessed the pattern of steatosis, craniocaudal liver span, caudate-to-right lobe ratio, preportal space distance, and presence of porta hepatis lymph nodes and ascites. Biopsy specimens were assessed by a pathologist, and the degree of necroinflammatory activity, steatosis, and fibrosis was determined. Histopathologic and CT findings were compared between patients with NASH and patients with steatosis alone using the Mann-Whitney U test and Fisher exact test. RESULTS: In patients with NASH, the mean liver-to spleen attenuation ratio was 0.66 (range: 0.1-1.1). Steatosis was diffuse (n=9), geographic or nonlobar (n=2), or diffuse with an area of focal sparing (n=1). The liver craniocaudal span varied from 17.5 to 25.5 cm (mean=21.4 cm), and hepatomegaly was present in 11 (91.7%) patients. The caudate-to-right-lobe ratio (mean=0.43) and preportal space (mean=4.5 mm) were normal in all cases. Porta hepatis lymph nodes were present in 7 (58.3%) patients; their mean dimensions were 16 mmx11 mm. Ascites was absent in all patients. On histopathology, the degree of necroinflammatory activity was mild (n=9), moderate (n=1), or severe (n=2). The degree of steatosis was 33% to 66% (n=5) or >67% (n=7). All but 3 patients had fibrosis; 6 had focal nonbridging fibrosis, 1 had multifocal nonbridging fibrosis, and 2 had bridging fibrosis. There was a significant correlation between the degree of steatosis on pathologic examination and the liver-to-spleen attenuation ratio on CT (P=0.048). The severity of inflammation and stage of fibrosis on pathologic examination did not correlate with the CT features. Among patients with steatosis alone, the mean liver-to-spleen attenuation ratio was 0.80 (range: 0.3-1.2); the craniocaudal liver span varied from 12 to 20 cm (mean=16 cm); hepatomegaly was present in 2 (22.2%) patients; the caudate-to-right lobe ratio was normal in all patients, with a mean of 0.36 (range: 0.22-0.47); the preportal space distance was enlarged in 2 cases (mean=7.5 mm, range: 1-16 mm); porta hepatis lymph nodes were present in 7 (77.8%) patients, and their mean dimensions were 11 mmx8 mm (large axis range: 6 19 mm, short axis range: 4-14 mm); and no patient had ascites. There was a significant difference in the craniocaudal liver span between patients with NASH (mean=21 cm) and patients with steatosis (mean=16 cm) (P<0.05). The caudate-to right-lobe ratio was also significantly different between patients with NASH (mean=0.43) and patients with steatosis (mean=0.36) (P<0.05). There were no significant differences in liver-to-spleen attenuation ratios, measurements of preportal space, or the presence of porta hepatic lymph nodes. CONCLUSION: The CT features of NASH include steatosis, hepatomegaly, and porta hepatis lymph nodes, and the liver-to-spleen attenuation ratio correlated with the degree of steatosis on histopathology. Patients with NASH had a greater liver span and increased caudate-to-right-lobe-ratio compared with patients with steatosis alone. PMID- 16365571 TI - Does ultra-low-dose CT with a radiation dose equivalent to that of KUB suffice to detect renal and ureteral calculi? AB - The objective of this study was to evaluate the diagnostic yield of multislice CT using a radiation dose equivalent to that of conventional abdominal x-ray (KUB). One hundred forty-two patients were prospectively examined with ultrasound and a radically dose-reduced CT protocol (120 kV, 6.9 eff. mAs). Number and size of calculi, presence of urinary obstruction, and alternative diagnoses were recorded and confirmed by stone removal/discharge or by clinical and imaging follow-up. The mean effective whole-body dose was 0.5 mSv in men and 0.7 mSv in women. The sensitivity and specificity in detecting patients with calculi was 97% and 95% for CT and 67% and 90% for ultrasound. Urinary obstruction was similarly assessed, whereas CT identified significantly more alternative diagnoses than ultrasound (P<0.001). With regard to published data for standard-dose CT, the present CT protocol seems to be comparable in its diagnostic yield in assessing patients with calculi, and its radiation dose is equivalent to that of KUB. PMID- 16365572 TI - Oral contrast adherence to polyps on CT colonography. AB - Although oral contrast agents are known to improve the accuracy of CT colonography (CTC) by tagging fluid and stool, it is not well recognized that oral contrast also adheres to the surface of polyps. The authors' objective was to quantitate the frequency of contrast adhering to polyps. Three hundred thirty eight optical colonoscopy-proven polyps were identified on CTC of all of the 216 patients with polyps in a larger cohort of screening patients. CT scans of polyps were analyzed for adherent contrast (ie, a thin coat/adherent drops) in at least one view (prone/supine). Forty-six percent of the 312 polyps not touching a contrast pool had adherent contrast. Polyps with villous histology were significantly more likely to have adherent contrast (77% [20/26] vs. 43% [124/286], P<0.001). Oral contrast agents often tag polyp surfaces in a pattern that is distinct from internal tagging of adherent stool, which must be recognized during CTC interpretation. Polyps with villous histology show a higher rate of contrast adherence than nonvillous polyps. PMID- 16365573 TI - Characterization of normal ileocecal valve density on CT colonography. AB - The objective of this study was to evaluate normal ileocecal valve (ICV) attenuation and composition on CT colonography (CTC). Two hundred twelve patients who underwent CTC at the authors' institution were retrospectively identified. Two independent readers recorded three region-of-interest (ROI) measurements, which were used to determine mean ICV density. ICV attenuation measurements were compared with age and abdominal body fat using Pearson correlation. A subjective classification system was applied to characterize the relative fat and soft tissue density of the ICV. Mean ICV density was -26.3+/-14 HU (range -60.1 to 18.3 HU). There was no correlation between the fat content of the ICV and age (r=0.06) or body fat (r=-0.47). The majority of valves (83.5%) were classified as heterogeneously low density or heterogeneously high density, whereas a small fraction of valves (6.1%) were categorized as homogeneously low density. The results of this study suggest that ICVs have a varying range of densities, and this feature alone cannot be used to distinguish the ICV from a polyp or neoplastic lesion on CTC. PMID- 16365574 TI - Magnetic resonance imaging findings of tuberculous endometritis: a report of 2 cases. AB - The magnetic resonance imaging findings in 2 patients pathologically proven to have endometrial tuberculosis are reported. PMID- 16365575 TI - Lymphadenopathy in the mesenteric pedicle of the free jejunal flap: reactive lymphadenopathy, not metastatic. AB - OBJECTIVE: Reconstructive surgery using the free jejunal flap is sometimes performed to close surgical defects in patients undergoing pharyngolaryngectomy for laryngeal or hypopharyngeal cancer. The clinical significance of lymph nodes in the flap was retrospectively examined. METHODS: Fifteen patients had undergone a laryngectomy with free jejunal flap reconstruction between March 1996 and October 1999. The appearance and size of lymph nodes of the flap were examined by 3 radiologists. RESULTS: Lymph nodes were observed in 10 patients. Size increases were noted within 1 year after surgery but not beyond 1 year. The pathologic diagnosis of the excised nodes was reactive lymphadenopathy. CONCLUSIONS: Lymph nodes may occasionally become apparent in the free jejunal flap. There was no suggestion of metastatic lymph nodes when examined retrospectively. We speculate that this reaction was the result of environmental changes attributable to surgery. PMID- 16365576 TI - Neurilemmoma of the glans penis: ultrasonography and magnetic resonance imaging findings. AB - Neurilemmoma of the glans penis is rare, and no imaging findings have been reported. A case of neurilemmoma of the glans penis is presented. Ultrasonography (US) and magnetic resonance imaging revealed a well-defined small mass in the glans penis. The mass appeared hypoechoic on gray-scale US and hypervascular on color Doppler US. Magnetic resonance imaging revealed high signal intensity of the mass on a T2-weighted image and strong enhancement on a contrast-enhanced T1 weighted image. PMID- 16365577 TI - Integrated PET/CT and the dry pleural dissemination of peripheral adenocarcinoma of the lung: diagnostic implications. AB - OBJECTIVE: The aim of this study was to describe retrospectively the CT findings of dry pleural dissemination of peripheral lung adenocarcinoma, and to compare the mutual roles of PET and CT components of integrated PET/CT in the diagnosis of the disease. METHODS: The authors analyzed retrospectively the CT findings of pathologically proved dry pleural dissemination in 8 of 172 patients with peripheral adenocarcinoma of the lung. Subsequently, one radiologist and one nuclear medicine physician (unaware of the CT and pathologic results) evaluated together in a random order the integrated PET/CT of 172 adenocarcinoma patients (8 with dry pleural dissemination and 164 without). They recorded the presence of pleural dissemination using PET images only and using both PET and CT images. The diagnostic accuracies with respect to the presence of pleural dissemination were evaluated. RESULTS: The CT findings of dry pleural dissemination were pleural small nodules (n=8, 100%) (>or=6 in number in all patients; 198/204 nodules were <5 mm in diameter and 6/204 were 5-10 mm) and uneven (n=4, 50%) or band-like (n=3, 38%) fissural thickening. By PET only, the sensitivity, specificity, and accuracy of dry pleural dissemination were 25% (2/8), 90% (147/164), and 87% (149/172), respectively; by PET plus CT these were 100% (8/8), 100% (164/164), and 100% (172/172), respectively. CONCLUSIONS: The CT findings of dry pleural dissemination are multiple small pleural nodules and uneven pleural thickening. Dry pleural dissemination should be diagnosed using CT findings at integrated PET/CT because lesions causing pleural dissemination without pleural effusion are usually beyond PET resolution. PMID- 16365578 TI - Detection of pulmonary embolism using 16-slice multidetector-row computed tomography: evaluation of different image reconstruction parameters. AB - OBJECTIVE: To compare different image reconstruction parameters for detecting emboli of the pulmonary arteries according to anatomic levels using 16-slice multidetector-row computed tomography in patients suspected of having an acute pulmonary embolism (PE). METHODS: Sixty-two patients (33 male and 29 female) with a clinically suspected acute PE were included in the present study. Multidetector row computed tomography scans were performed using 16-mmx0.75-mm collimation. Based on the computed tomography data set, different image reconstruction parameters were used for each patient: axial slice thicknesses (STs) of 0.75, 2, 4, and 6 mm; axial maximum intensity projection (MIP) STs of 4 mm; and coronal STs of 2 and 4 mm. In joint reading fashion, 2 experienced radiologists reviewed examination findings regarding the presence and/or absence of a PE. The reference standard for visualization and detection of PEs was defined using the axial images with a 0.75-mm ST. RESULTS: In 23 of 62 patients, a PE was diagnosed. For main and lobar pulmonary arteries, the sensitivities and specificities were as follows: axial 2-mm images, 1.0/1.0; axial 4-mm images, 1.0/1.0; axial 6-mm images, 0.97/0.99; MIP 4-mm images, 0.95/0.99; coronal 2-mm images, 1.0/1.0; and coronal 4-mm images, 1.0/1.0. Regarding segmental and subsegmental pulmonary arteries, sensitivity and specificity varied: axial 2-mm images, 0.97/1.0; axial 4-mm images, 0.81/0.99; axial 6-mm images, 0.65/0.99; axial MIP 4-mm images, 0.63/0.99; coronal 2-mm images, 0.91/0.99; and coronal 4-mm images, 0.74/0.99. CONCLUSIONS: In detecting segmental and subsegmental PEs, only the axial images with an ST of 2 mm proved to have results comparable with the axial 0.75-mm images. Therefore, thin-slice collimation and ST are mandatory for visualization of segmental and subsegmental PEs in patients suspected of having an acute PE. PMID- 16365579 TI - Comprehensive assessment of lung CT attenuation alteration at perfusion defects of acute pulmonary thromboembolism with breath-hold SPECT-CT fusion images. AB - Regional computed tomography attenuation (CTA) alteration at perfusion defects in acute pulmonary thromboembolism (PTE) was comprehensively assessed using deep inspiratory breath-hold SPECT-CT fusion images. Subjects were 14 acute and 9 chronic PTE patients and 13 control subjects. Regional perfusion, CTA, and intravascular clots were correlated on deep-inspiratory breath-hold SPECT unenhanced/angiographic CT fusion images. Fusion images visualized hypo-CTA in 57% of the acute PTE patients, which preferentially occurred at extensively and severely decreased perfusion areas caused by central clots. CTA at 35 defects of acute PTE was significantly decreased compared with that of normal lungs (P<0.001), but the degree was less compared with chronic PTE (P<0.0001). Fusion images also revealed variable relationships of clots and regional perfusion/CTA in the distal lungs of each central clot. Fusion images provide important information about the actual effects of intravascular clots on peripheral perfusion/CTA and indicate that lung CTA can be decreased at perfusion defects in acute PTE. PMID- 16365580 TI - Frequent cause of the Macklin effect in spontaneous pneumomediastinum: demonstration by multidetector-row computed tomography. AB - OBJECTIVE: The purpose of this study was to evaluate the Macklin effect in patients with spontaneous pneumomediastinum by multidetector-row computed tomography (MDCT). METHODS: Chest computed tomography (CT) reports between July 2000 and January 2003 at our institution were retrospectively reviewed. Nine patients with spontaneous pneumomediastinum were identified, and their CT scans were evaluated to detect the Macklin effect. The patient diagnoses included interstitial lung diseases (n=4), bronchiolitis obliterans organizing pneumonia or pneumonia (n=3), asthma (n=1), and spontaneous pneumomediastinum without definite pulmonary disease (n=1). RESULTS: The Macklin effect was observed in 8 (89%) of 9 patients. In 4 patients, the Macklin effect was observed in perihilar and peripheral areas, and in 4 patients, it was observed only in the perihilar area. CONCLUSIONS: The Macklin effect can frequently be demonstrated in patients with spontaneous pneumomediastinum of nontraumatic respiratory causes by MDCT. A CT-proven Macklin effect may be useful in differentiating respiratory from other causes of pneumomediastinum. PMID- 16365581 TI - High-resolution computed tomography findings of lung cancer associated with idiopathic pulmonary fibrosis. AB - OBJECTIVE: The purpose of this study was to evaluate high-resolution computed tomography (HRCT) findings of lung cancer associated with idiopathic pulmonary fibrosis (IPF). METHODS: Thirty patients with lung cancer who had preceding IPF and were receiving regular follow-up between 1993 and 2002 were examined. Medical records, radiographs (including HRCT scans), and histologic slides were reviewed. RESULTS: In 28 of the 30 patients, the most common HRCT pattern of lung cancer was a nodular lesion with soft tissue attenuation. Nodule margins were well defined in 23 lesions (82.1%), associated with lobulation in 24 (85.7%), or characterized by spiculation in 14 (50%). Air bronchogram was observed in 16 lesions (57.1%). All nodules were located in the peripheral area of fibrotic lesions. Squamous cell carcinoma and adenocarcinoma were the most frequent histologic types. CONCLUSIONS: The typical HRCT findings of lung cancer were well defined nodular lesions with lobulation in peripheral areas of the lung. PMID- 16365582 TI - Multimodality diagnostic imaging of diffuse esophageal leiomyomatosis. AB - A case of diffuse esophageal leiomyomatosis is presented, with emphasis on the imaging findings across multiple contemporary diagnostic modalities. This entity represents a rare presentation of uncommon benign smooth muscle tumors of the esophagus. The characteristic clinical, histologic, and multimodality imaging findings may distinguish this benign tumor from its malignant counterpart leiomyosarcoma and from achalasia. PMID- 16365583 TI - Improvement of sensitivity and interrater reliability to detect acute stroke by dynamic perfusion computed tomography and computed tomography angiography. AB - OBJECTIVE: To assess the benefits of additional computed tomography perfusion (CTP) and computed tomography angiography (CTA) on the detection of early stroke, vessel occlusion, estimated infarct size, and interrater reliability. METHODS: Sixty-seven consecutive patients underwent nonenhanced computed tomography (CT) imaging, CTA, and CTP. The final diagnosis of stroke was made from follow-up neuroimaging. A first diagnosis was made on-site by the physician on duty. Three experienced neuroradiologists blinded to follow-up findings analyzed the data set off-line, evaluated CT for signs of acute stroke, and subsequently evaluated CTP and CTA for infarction-related perfusion deficits and vessel abnormalities. RESULTS: Computed tomography perfusion and CTA increased the time from CT start to diagnosis from 2 minutes to 10 minutes. Sensitivity to detect acute stroke increased significantly in all investigators from 0.46-0.58 to 0.79-0.90 compared with CT (<0.005). The interrater weighted kappa value increased from 0.35 to 0.64. Estimation of infarct size was not improved. CONCLUSION: Computed tomography perfusion and CTA provide an effective add-on to standard CT in acute stroke imaging by significantly increasing the sensitivity and reliability of infarct detection. PMID- 16365584 TI - Visualization of spinal cord motion associated with the cardiac pulse by tagged magnetic resonance imaging with particle image velocimetry software. AB - OBJECTIVE: The purpose of this study was to evaluate whether or not tagged magnetic resonance (MR) imaging with particle image velocimetry (PIV) software could reveal spinal cord motion clearly. METHODS: Six volunteers were enrolled in this study. Tagged MR imaging using fast spoiled gradient-recalled acquisition in the steady state with spatial modulation of the magnetization technique was performed using a 1.5-T MR system. RESULTS: Sagittal vector maps analyzed by PIV software revealed entire spinal cord motion sequentially during the cardiac cycle. The cervical spinal cord initially moved in a caudal direction and then continually oscillated from a cephalic-to-caudal direction. Each volunteer had a different cycle. In the thoracic spinal area, similar findings were observed, although they were slightly less clear than in the cervical area. CONCLUSION: Tagged MR imaging combined with PIV software, referred to as tagged MR image velocimetry, revealed spinal cord motion associated with the cardiac pulse, especially in the cervical spine. PMID- 16365585 TI - Brain stem and cerebellar findings in Joubert syndrome. AB - Joubert syndrome is often missed clinically and radiologically if not enough attention is paid to its subtle and variable clinical presentation and the imaging findings in the posterior fossa. The purpose of this paper is to illustrate the brain stem and cerebellar imaging findings in Joubert syndrome. Awareness of the clinical and neuroimaging findings in Joubert syndrome and maintenance of a high index of suspicion are essential to correctly diagnose this rare congenital malformation. PMID- 16365586 TI - Sinonasal computed tomography in patients with Wegener's granulomatosis. AB - The goal of this study was to describe pathologies of sinonasal CT in patients with Wegener's granulomatosis. Between 1993 and 2004, sinonasal CT was performed in 28 patients (15 male, 13 female) with Wegener's granulomatosis. The following imaging findings were assessed on the CT scans: mucosal thickening, subtotal opacification, air-fluid level, bony destruction, sclerosing osteitis, bony thickening, orbital mass, and saddle nose deformity. Of the 28 patients, 61% showed mucosal thickening in the nasal cavity and 75% in the paranasal sinuses. A subtotal opacification of the paranasal sinuses was detected in 25%, and of the mastoid cells in 7% of patients. Fifty-seven percent of patients had bony destruction of the nasal cavity, and 54% of the paranasal sinuses. CT revealed sclerosing osteitis of the paranasal sinuses in 21%, and of the mastoid cells in 18% of patients. Bony thickening of the paranasal sinuses was detected in 18%, and of the mastoid cells in 14% of patients. With respect to all imaging findings, the maxillary sinuses were the most frequently affected paranasal sinuses. The authors conclude that the following sinonasal CT findings are observed in patients with Wegener's granulomatosis: (1) bony destruction, mainly of the nasal cavity, maxillary sinuses, and mastoid cells; (2) sclerosing osteitis, mainly of the maxillary sinuses and mastoid cells; (3) bony thickening, mainly of the maxillary sinuses and mastoid cells; and (4) mucosal thickening, mainly of the maxillary sinuses. These findings are, however, nonspecific and should be combined with other clinicopathologic and laboratory studies to confirm the diagnosis of Wegener's granulomatosis. PMID- 16365587 TI - Early detection of subacute sclerosing panencephalitis by high b-value diffusion weighted imaging: a case report. AB - The authors report a case of subacute sclerosing panencephalitis in which the diagnosis was suggested by high b-value diffusion-weighted imaging (DWI) findings. The signal abnormalities were located asymmetrically at bilateral cerebral corticosubcortical regions. High b-value DWI showed these signal abnormalities as marked hyperintensity with decreased apparent diffusion coefficient values. The signal abnormalities were difficult to identify on other magnetic resonance imaging sequences, including routine DWI. High b-value DWI could be valuable for earlier detection of subacute sclerosing panencephalitis. PMID- 16365588 TI - Isolated hypoplasia of the tentorium. AB - Developmental defects of the tentorium represent a spectrum of disease with small defects and apertures that have little clinical significance on one end and hypoplasia or aplasia on the other. The latter is associated with serious central nervous system malformations, it is seldom isolated, and aside from the 3 cases described below, there have only been 2 case reports published on isolated hypoplasia. This condition is important to recognize so as to avoid unnecessary investigations and interventions. PMID- 16365589 TI - A prion disease--possible Gerstmann-Straussler-Scheinker disease: a case report. AB - A 50-year-old patient with a 6-month history of progressive cognitive and motor disability is presented. There were no myoclonic jerks on examination and no periodic sharp waves by electroencephalography. Imaging showed high signal on T2 weighted scans in the basal ganglia and posterior limbs of the internal capsules, with no restricted diffusion and parenchymal volume loss. A brain biopsy was performed. Western blot analysis revealed a protease-resistant prion protein fragment (PrP7-8), the molecular hallmark of Gerstmann-Straussler-Scheinker disease. PMID- 16365590 TI - Overall breast density in MR mammography: diagnostic and therapeutic implications in breast cancer. AB - The objective of this study was to assess the efficacy of MR mammography (MRM) in evaluating breast cancer extent in women with fatty or dense breasts, and its contribution to the therapeutic approach. The authors reviewed 97 carcinomas detected in 93 women (both symptomatic and from screening) that were classified in two groups according to breast density pattern. Mammography, ultrasound (US), and MRM were performed to evaluate size, extension of the in situ component, presence of multifocal/multicentric disease, and contralateral involvement. Results obtained on mammography plus US were balanced against MRM, considering pathologic analysis as the gold standard. For fatty breasts (n=47), exact measurement was found on mammography plus US and on MRM alone in 70%, underestimation on mammography plus US 23.5% and on MRM 11% (P=0.005). For dense breasts (n=50), exact measurement was found on mammography plus US in 40% and on MRM alone 68%, underestimation on mammography plus US 52% and on MRM 10% (P=0.005). Overall, good correlation (R>0.71) was found between pathologic and clinical size with all imaging methods; nevertheless, when evaluating multifocal/multicentric disease, a poor correlation was observed between histologic assessment and mammography plus US (R=0.52), but it was excellent with regard to MRM (R=0.99). In fatty breasts, the combination of mammography and US allows for a precise assessment of tumoral extension. However, these results show that in dense breasts, MRM is superior to mammography plus US, suggesting that its systematic use in this group of patients is justifiable. PMID- 16365591 TI - Adenoma of the nipple: correlation of magnetic resonance imaging findings with histologic features. AB - Adenoma of the nipple (nipple adenoma) is a rare breast tumor that can show various histologic features. We present magnetic resonance (MR) and other imaging findings of a case of nipple adenoma and correlate them with detailed histologic features of the tumor. The lesion showed early strong enhancement with rim enhancement on dynamic contrast-enhanced MR images, and internal portions of the lesion showed washout on delayed MR images. Histologically, the tumor showed expansive growth with surrounding dense collagenous tissue and large vessels. Internal portions of the lesion showed a relatively low degree of fibrosis compared with that of the surrounding area, and tiny vessels were observed. These features were clearly reflected in the MR findings. PMID- 16365592 TI - Value of whole-body turbo short tau inversion recovery magnetic resonance imaging with panoramic table for detecting bone metastases: comparison with 99MTc methylene diphosphonate scintigraphy. AB - OBJECTIVE: The objectives of this study were to assess the efficacy and reliability of whole-body turbo short tau inversion recovery (STIR) magnetic resonance imaging (MRI) for detecting skeletal metastasis and to compare the results with those of bone scintigraphy. METHODS: Twenty-six patients with primary cancer (mean age=56 years, age range: 34-75 years) were assessed for bone metastasis with whole-body MRI and bone scintigraphy. Eight bone regions in each patient were assessed (total of 208 sites) with each of these 2 techniques. A turbo STIR sequence and panoramic table were used during MRI. Whole-body MRI and scintigraphy findings were compared with biopsy or follow-up imaging results. RESULTS: After at least 12 months of follow-up, 9 patients had bone metastases in a total of 31 sites. Whole-body MRI showed 29 metastases (94%) in the total 208 skeletal sites investigated in the 26 patients. Bone scintigraphy revealed metastases in 16 (52%) of the 208 sites. CONCLUSION: Whole-body turbo STIR MRI is a reliable method for screening patients with suspected skeletal metastases. This technique is also advantageous in that it reveals extraskeletal organ and soft tissue metastases. PMID- 16365593 TI - Hemodialysis fistula occlusion: demonstration with 64-slice CT angiography. AB - The speed and resolution of 64-slice CT have resulted in new applications for CT angiography (CTA) owing to rapid data acquisition during the arterial phase, improved visualization of small vessels, and lengthened anatomic coverage. Extremity CT angiography is one such region. This case report shows the utility of multislice CTA for the evaluation of hemodialysis graft dysfunction. PMID- 16365594 TI - Sectional neuroanatomy of the middle thoracic spine (t5-t8) and chest. AB - This is the third in a series of articles on the spine. The first reviewed the anatomy of the neck. The second reviewed the upper thoracic spine and chest (T1 T4). A fourth article is planned for the lower thoracic spine. Procedures in the midthoracic spine include chest tube placement, trigger point injections, chemodenervation with botulinum toxin, video-assisted thoroscopic surgery, and spinal injections. Complications include pneumothorax, hemothorax, diaphragmatic irritation, sympathetic trunk irritation, postthoracotomy pain, and intradural abscesses. This article provides anatomically accurate schematics of innervations of the middle thoracic chest and spine (T5-T8) that can be used to interpret magnetic resonance images of the muscles and nerves. Cross-sectional schematics of the middle thoracic chest and spine were drawn as they appear on imaging projections. The relevant nerves were color coded. The muscles and skin surfaces were labeled and assigned the color of the appropriate nerves. An organized comprehensive map of the motor innervation of the middle thoracic chest and spine allows the physician to increase the accuracy and efficacy of interventional procedures. This could also assist the electromyographer in correlating the clinical and electrophysiologic findings with magnetic resonance images. PMID- 16365595 TI - Fully human therapeutic monoclonal antibodies. AB - Monoclonal antibody (mAb) therapy has been facilitated by a number of technologic advances over the past 30 years. Whereas hybridoma development of murine mAbs was requisite for the development of mAbs as drugs, the inherent immunogenicity of rodent sequences in humans has presented obstacles to the clinical application of mAbs. Sensitization to mAb therapeutics poses significant risk to the patient and may blunt the efficacy of these therapies. The advent of chimeric antibodies lessened but did not eliminate the rodent content of mAbs; thus, immunogenicity remained a concern. Further elimination of rodent sequences enabled the production of humanized mAbs, followed by current technology using phage display and, finally, transgenic mice technology, which allows for the generation of fully human therapeutic mAbs. The reduced immunogenicity of this new generation of mAbs is expected to enhance efficacy, safety, and ease of use. In addition to providing replacements for existing mAb drugs, new technologies have greatly facilitated the optimization and modification of mAbs, opening numerous therapeutic avenues. PMID- 16365596 TI - IL-12 + GM-CSF microsphere therapy induces eradication of advanced spontaneous tumors in her-2/neu transgenic mice but fails to achieve long-term cure due to the inability to maintain effector T-cell activity. AB - A single intratumoral injection of interleukin-12 and granulocyte-macrophage colony-stimulating factor-encapsulated microspheres induced the regression of advanced spontaneous mammary tumors, suppressed additional tumor development, and enhanced survival in her-2/neu transgenic mice. Posttherapy tumor eradication was dependent on both CD4+ and CD8+ T cells and correlated with the tumor infiltration kinetics of a transient effector T-cell response. Upon long-term monitoring, tumor regression was found to be temporary, and disease-free survival was not achieved despite the development of systemic anti-tumor cytotoxic T-cell memory and antibody responses. Repeated immunization of mice enhanced short-term tumor suppression, resulting in the complete regression of primary tumors in up to 40% of the mice, but did not improve long-term survival owing to recurrence. The failure of chronic therapy to achieve complete cure was associated with an inability to maintain the intensity of the posttherapy effector T-cell response in this model. PMID- 16365597 TI - Target antigen expression on a professional antigen-presenting cell induces superior proliferative antitumor T-cell responses via chimeric T-cell receptors. AB - Human T cells expressing tumor antigen-specific chimeric receptors fail to sustain their growth and activation in vivo, which greatly reduces their therapeutic value. The defective proliferative response to tumor cells in vitro can partly be overcome by concomitant CD28 costimulatory signaling. We investigated whether T-cell activation via chimeric receptors (chRec) can be further improved by ligand expression on antigen-presenting cells of B-cell origin. We generated Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTLs) expressing a CD19-specific chRec. These CTLs are provided with native receptor stimulation by autologous EBV-transformed B-lymphoblastoid cell lines (LCLs) but exclusively with chRec (CD19-specific) stimulation by allogeneic, human leukocyte antigen (HLA)-mismatched CD19+ LCLs. CD19zeta-transduced EBV specific CTLs specifically lysed both allogeneic EBV targets and CD19+ tumor cells through the chRec in a major histocompatibility complex-independent manner, while maintaining their ability to recognize autologous EBV targets through the native T-cell receptor. The transduced CTLs failed to proliferate in response to CD19+ tumor targets even in the presence of CD28 costimulatory signaling. By contrast, CD19 expressed on HLA-mismatched LCL-induced T-cell activation and long term proliferation that essentially duplicated the result from native receptor stimulation with autologous LCLs, suggesting that a deficit of costimulatory molecules on target cells in addition to CD28 is indeed responsible for inadequate chRec-mediated T-cell function. Hence, effective tumor immunotherapy may be favored if engagement of the chRec on modified T cells is complemented by interaction with multiple costimulator molecules. The use of T cells with native specificity for EBV may be one means of attaining this objective. PMID- 16365598 TI - Identification of H-2Db-specific CD8+ T-cell epitopes from mouse VEGFR2 that can inhibit angiogenesis and tumor growth. AB - Vascular endothelial growth factor receptor 2 (VEGFR2/KDR) plays a crucial role in tumor-associated angiogenesis and vascularization. It has been established that monoclonal antibodies against VEGFR2 can inhibit angiogenesis. In this study, two naturally processed CD8 T-cell epitopes (VILTNPISM and FSNSTNDILI) were identified from murine KDR. Cytotoxic T lymphocytes targeting endothelial cells could be directly monitored by KDR2 and KDR3 Elispots or major histocompatibility complex class I tetramer staining. Immunization with these two peptides effectively reduced angiogenesis and inhibited tumor growth in mouse models. Thus, vaccination with KDR peptides alone or in combination with other anti-angiogenesis agents may afford a novel immunotherapy for inhibition of tumor growth. PMID- 16365599 TI - Monoclonal antibodies targeted against melanoma and ovarian tumors enhance dendritic cell-mediated cross-presentation of tumor-associated antigens and efficiently cross-prime CD8+ T cells. AB - Dendritic cells (DCs) constitute very attractive vectors for cancer immunotherapy due to their ability to efficiently capture and present tumor antigens, which initiates tumor-directed T-cell responses. Because the initiation of cytotoxic anti-tumor immune responses requires the cross-presentation mechanism, antigen targeting to DCs represents a very important step in the chain of events that constitutes the cross-priming immune process. In the current study, we explored the ability of DCs loaded with antibody-coated melanoma and ovarian carcinoma tumor cells to cross-present tumor antigens to CD8+ T cells and elicit in vitro anti-tumor immune responses. Coating melanoma and ovarian cancer cells with monoclonal antibodies against different surface antigens (CD44, ME491, LFA-3, and CD24) expressed by the tumor cells promoted the cross-presentation of the tumor associated antigens as MART-1, gp100, tyrosinase, and NY-ESO-1 by DCs to CD8+ T. These tumor antigen-specific CD8+ T-cell populations resulting from the DC mediated cross-priming process were identified using specific immune tetramers and were a few fold larger than the ones generated using peptide-pulsed or apoptotic tumor cell-loaded DCs. The CD8+ T cells generated by DCs loaded with monoclonal antibody-coated tumor cells were cytotoxic against the primary melanoma and ovarian carcinoma cells. Thus, targeting monoclonal antibody-coated tumor cells to DCs is a novel method that opens new perspectives for immunotherapy strategies. PMID- 16365600 TI - Long-term preservation of antibody-dependent cellular cytotoxicity (ADCC) of natural killer cells amplified in vitro from the peripheral blood of breast cancer patients after chemotherapy. AB - Twenty percent of breast cancer adenocarcinomas overexpress the oncogene c-erb-2 that is recognized by the humanized anti-Her2/neu monoclonal antibody Herceptin. Results from clinical studies suggest that antibody-dependent cellular cytotoxicity (ADCC) is involved in the clinical response of Herceptin-treated patients. The purpose of the current study was to evaluate the possibility of amplifying in vitro the CD3-/CD16+ natural killer (NK) cell subset that mediates ADCC from breast cancer patients after chemotherapy. Peripheral blood mononuclear cells from six breast cancer patients taken 2 months after chemotherapy completion were co-cultured with an autologous irradiated Epstein-Barr virus transformed B-lymphoblastoid cell line (LCL) in the presence of interleukin-2 (IL 2) for 4-6 weeks. These LCL + IL2 activated cultures (ACs) were tested for ADCC potential, and their CD3/CD16 NK proportion was quantified. Among the ACs, the proportion of CD3-/CD16+ NK cells increased up to 64% over the first 2 weeks of culture and the ACs continued to expand for 1 month thereafter. Control and patient ACs displayed ADCC activity (tested in the presence of Rituximab against the autologous LCL to take into account any possible effect of inhibitory NK receptors) as well as against the MCF-7(Her2/neu) breast cancer cell line in the presence of Herceptin. This ADCC activity was maintained during the entire culture period. In conclusion, chemotherapy in breast cancer patients does not obviate the possibility of amplifying in vitro the NK cell subset that mediates ADCC. Consequently, adoptive transfer of lymphocytes mediating ADCC can be considered using this protocol to test its benefit in patients under Herceptin treatment. PMID- 16365601 TI - Intraclonal variability of VH genes in follicular lymphoma patients who have received anti-idiotypic immunotherapy. AB - Subclonal heterogeneity can affect idiotypic determinants present in the clonotypic immunoglobulin of B-cell follicular lymphomas (FLs) and may limit the effect of antilymphoma treatments performed by immunization of patients with their own tumor-associated idiotypic immunoglobulin. Idiotype-secreting hybridomas were obtained by fusion of tumor cells from 5 patients with FL, and the K6H6/B5 human heteromyeloma and rearranged VH genes from tumor samples and hybridomas were amplified, cloned, and sequenced. Sequences were aligned with germline genes and somatic mutations, intraclonal heterogeneity and genealogic relations of the B-cell clones in the different biopsy specimens were determined. The VH sequence of the progenitor clone was determined in samples of the tumoral population. Further diversification resulted in the presence of 2 to 6 subclones in 4 of the 5 samples studied. Only in 1 patient did the hypermutation mechanism introduce differences among most of the potential idiotopes present in individual subclones. The VH sequence of the hybridoma that provided the idiotypic-vaccine was identified in one of the tumor subclones in all cases. No relapse has been demonstrated in 3 of the 4 vaccinated patients (follow-up: 29-103 months). We conclude that despite potential differences in the idiotypic region expressed by individual tumor cells, at least some potential idiotopes may be preserved among all the tumor subclones in most cases studied. All vaccinated patients developed immune responses against the autologous tumor idiotypic immunoglobulin. Polyclonal anti-idiotypic immune responses induced with a vaccine obtained from 1 hybridoma may be effective against all the idiotypic variants present in the tumor population. PMID- 16365602 TI - Quick generation of fully mature dendritic cells from monocytes with OK432, low dose prostanoid, and interferon-alpha as potent immune enhancers. AB - Dendritic cells (DCs) are one of the promising tools for enhancing antigen specific immune responses in clinical settings. Many studies have been performed thus far to verify the efficacy of the DC vaccine in cancer patients; however, the responses have not always been satisfactory, partly because of DC incompetence. To obtain DCs potentially applicable for vaccination of cancer patients, our group sought to establish the strategy of DC generation mainly by modulating culture periods and maturation stimuli. Novel mature DCs that can be generated from monocytes within 3 days by using a combination of OK432 (Streptococcus pyogenes preparation), low-dose prostaglandin E2 (PGE2), and interferon-alpha (OPA-DCs) were developed. They strongly express CD83, CD86, and CCR7 and have potent ability to migrate to CCL21. In addition, they were able to activate natural killer and T helper 1 (TH1) cells and to induce peptide-antigen specific cytotoxic T lymphocytes more significantly than monocyte-derived DCs stimulated with a conventional cytokine cocktail of tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, and PGE2 (monocyte-conditioned medium [MCM]-mimic DCs). The profound ability of OPA-DCs to stimulate these effectors is attributable to their higher expression of IL-12p70, IL-23, and IL-27 than MCM mimic DCs, which was supported by the findings that the neutralization of IL 12p70 and IL-23 reduced the TH1 priming ability of OPA-DCs. Even when from advanced gastric or colonic cancer patients, OPA-DCs displayed abilities of migration and TH1 induction comparable to those from healthy subjects. Therefore, OPA-DCs may serve as a feasible vaccine with the potential to enhance TH1 dominant and cytolytic immune responses against cancers. PMID- 16365603 TI - Mechanism of anticancer host response induced by OK-432, a streptococcal preparation, mediated by phagocytosis and Toll-like receptor 4 signaling. AB - It has previously been reported by our group that Toll-like receptor (TLR) 4 is involved in anticancer immunity induced by OK-432, a Streptococcus-derived immunotherapeutic agent. However the detailed mechanism of the OK-432-induced immune response via TLR4 remained uncertain, because it may not be possible for OK-432, which consists of whole bacterial bodies, to bind directly to TLR4. In the current study, we conducted in vitro and in vivo experiments to investigate the hypothesis that OK-432 may first be captured and dissolved by phagocytes and that the active components released by the cells may then induce host responses via TLR4. TS-2 monoclonal antibody, which recognizes an active component of OK 432 designated OK-PSA was used in the current study. First, it was observed that OK-432-induced cytokine production by dendritic cells (DCs) and macrophages was significantly inhibited in vitro by cytochalasin B, a phagocytosis inhibitor. Immunofluorescence staining using TS-2 demonstrated that OK-432 was captured and dissolved by phagocytes. OK-PSA was detected in the supernatants derived from OK 432-treated DC culture by enzyme-linked immunosorbent assay using TS-2. Supernatants from OK-432-treated DC culture increased nuclear factor (NF)-kappaB activity in TLR4-expressing cells, and the increased activity was inhibited by TS 2 antibody. OK-432 itself did not activate NF-kappaB in these cells. In in vivo experiments, the anticancer effect of OK-432 was significantly inhibited by suppression of phagocytosis activity by cytochalasin B. In this case, the amount of OK-PSA, an active component of OK-432, in the sera was also reduced by cytochalasin B. These findings elucidated the mechanism mediated by phagocytosis and TLR4 signaling in the immune effect of OK-432. PMID- 16365604 TI - Multicenter phase 2 study of interleukin-2 and 13-cis retinoic acid as maintenance therapy in advanced non-small-cell lung cancer. AB - High serum levels of vascular endothelial growth factor (VEGF) are a poor prognostic factor for patients with advanced non-small-cell lung cancer (NSCLC). We have previously shown that low-dose interleukin (IL)-2 and 13-cis retinoic acid (RA) decreased VEGF and improved the immune function of patients with advanced tumors treated with chemotherapy. The primary end point of this study was to verify whether IL-2 and RA decreased serum VEGF in NSCLC patients showing a clinical benefit from chemotherapy. The secondary end point was the evaluation of clinical outcome. We treated 38 patients with advanced NSCLC who had a complete or partial response or disease stability to chemotherapy and had a median serum VEGF level of 508 ng/mL; as maintenance therapy, they received subcutaneous IL-2 (1.8 x 10(6) IU) and oral RA. Matched controls (n = 87) were selected from a large cohort of patients with a similar disease status, including clinical benefit from chemotherapy. The most common adverse events were mild cutaneous skin rash and fever. Serum VEGF decreased to a mean level of 152 ng/mL (P = 0.0002). A statistically significant improvement in immune function was observed (lymphocyte and natural killer cell numbers and CD4+/CD8+ ratio) with respect to baseline values and controls. An improvement in the clinical outcome was also observed compared with controls. These data show that the administration of low-dose subcutaneous IL-2 and oral RA to patients with advanced NSCLC showing a clinical benefit from chemotherapy is feasible with a low-toxicity profile, decreases VEGF, and seems to improve progression-free and overall survival. PMID- 16365605 TI - Phase 2 study of the g209-2M melanoma peptide vaccine and low-dose interleukin-2 in advanced melanoma: Cancer and Leukemia Group B 509901. AB - High-dose interleukin-2 (IL-2) is the only approved immunologic therapy for advanced melanoma, but response rates are low and significant toxicities limit treatment to otherwise healthy patients. g209-2M is a nanopeptide engineered to mimic an epitope of the gp100 melanocyte differentiation protein that is recognized in a human leukocyte antigen (HLA)-restricted manner by melanoma tumor infiltrating lymphocytes in some patients. Previous reports indicated that administration of the g209-2M peptide could induce g209-reactive circulating T cells in patients with melanoma and that the combination of g209-2M and high-dose IL-2 might be a more active treatment than high-dose IL-2 alone. Low-dose IL-2 is not active but has significant biologic effects, and because of a different toxicity profile, it can be offered to most patients. The primary objective of this cooperative group phase 2 study was to determine the activity of the combination of g209-2M and low-dose IL-2 in advanced melanoma. Twenty-six HLA appropriate patients with advanced melanoma received subcutaneous g209-2M peptide once every 3 weeks and subcutaneous IL-2 (5 million IU/m) daily for 5 days during the first and second weeks. Patients were monitored for tumor response, toxicity, and induction of g209-reactive circulating T cells. There were no objective responses. There were no toxic deaths and no grade 4 toxicities. More than half of the patients experienced some grade 2 toxicity and one quarter experienced grade 3 toxicity. There was no convincing evidence by enzyme-linked immunospot or tetramer analysis of induction of g209-reactive circulating T cells. The combination of g209-2M and low-dose IL-2 is safe and tolerable but inactive against advanced melanoma. Absence of evidence of immunization raises concerns for peptide-based immunization strategies with concurrent IL-2. PMID- 16365606 TI - Profiling providers on use of adjuvant chemotherapy by combining cancer registry and medical record data. AB - PURPOSE: Treatment information collected by cancer registries can be used to monitor the provision of guideline-recommended chemotherapy to colorectal cancer patients. Incomplete information may bias comparisons of these rates. We developed statistical methods that combine data from a registry and physicians' records to assess hospital quality. DATA: From California Cancer Registry data, we selected all patients (n=12,594) newly diagnosed with stage III colon cancer or stage II or III rectal cancer from 428 hospitals during the years 1994 to 1998. To assess rates and predictors of underreporting of chemotherapy, we surveyed physicians treating 1449 of these patients from 98 hospitals during the years 1996 to 1997. METHODS: Using Bayesian statistical models, we imputed unobserved treatments. We studied the impact of underreporting on provider profiling by comparing rankings, estimates, and credible intervals based only on registry data to those incorporating physician survey data. RESULTS: Analyses that account for incompleteness of reporting yielded wider credible intervals for provider profiles than those that ignored such incompleteness. Among the 109 (25%) hospitals in the highest quartile of chemotherapy rates according to the registry data, 16 were not so classified when incomplete reporting was taken into account. With the more comprehensive model, 12 hospitals could be identified that ranked in the top quartile with probability>0.90. CONCLUSION: Estimates of adjusted hospital chemotherapy rates based solely on cancer registry data overstate the precision of assessments of hospital quality. Using additional information from a physician survey and applying rigorous statistical models, better inferences can be drawn about provider quality. PMID- 16365607 TI - Do mechanisms that link addiction treatment patients to primary care influence subsequent utilization of emergency and hospital care? AB - BACKGROUND: Patients with drug use disorders are heavy users of emergency department (ED) and inpatient hospital care. This study examines whether formal mechanisms to link addiction treatment patients to primary medical care, either directly on site or by off-site referral-when compared with an absence of said mechanisms-might reduce these patients' use of ED and hospital services after substance abuse treatment. METHODS: We used longitudinal data from 6 methadone maintenance programs with 232 patients, 24 outpatient nonmethadone programs with 1202 patients, and 14 long-term residential programs with 679 patients in the National Treatment Improvement Evaluation Study. Multivariate logistic models controlling for health status and medical service utilization before treatment examined whether provision of medical services on- or off-site during treatment linkage led to reduced use of ED and hospital services in the year after treatment compared with no such provision. RESULTS: On-site delivery of primary care reduced subsequent ED and hospital use among patients in methadone maintenance and long-term residential compared with the nonlinkage condition but not in outpatient nonmethadone programs. Off-site referral for medical care reduced subsequent ED visits but not hospitalizations in long-term residential programs. CONCLUSIONS: These findings suggest that for some treatment modalities, stronger primary care linkage mechanisms decrease subsequent utilization of expensive ED and hospital services. Future study should examine the cost implications of these strong linkage mechanisms and ways to strengthen linkages to off-site medical care. PMID- 16365608 TI - Income disparities in the quality of life of cancer survivors. AB - BACKGROUND: Studies of cancer survivors usually report positive correlations between income and health-related quality of life (QoL). These correlations cannot necessarily be interpreted as income disparities because earnings and income are affected by health, as well as the reverse. OBJECTIVES: The goal of this study was to quantify income disparities in QoL among cancer survivors by using instrumental variables (IV) to assess and, if necessary, correct for reverse causality. METHODS: We constructed an instrumental variable for income from home ownership, sources of unearned income, marital status at diagnosis, and spousal characteristics. Then, we examined income's effect on QoL in regressions controlling for other clinical and demographic predictors of QoL. The data were from interviews in 2002 with a cohort of cancer survivors who were 25 to 62 years of age when diagnosed during 1997 to 1999. MEASURES: The Functional Assessment of Cancer Therapy-General (FACT-G) and the SF-12 measured QoL in multiple domains. Questions adapted from the Health and Retirement Study ascertained the ratio of annual family income to the poverty threshold in 2001. RESULTS: Endogeneity tests were sensitive to assumptions of linearity for the income-QoL relationship and the choice of QoL measure. Consistently estimated income disparities were significant in all QoL models. The income elasticity of QoL ranged from 2% to 10%. CONCLUSIONS: There are income-related disparities in the QoL of cancer survivors that cannot be explained away by the effect of health on earnings. High income patients are not only more likely to survive cancer, but they enjoy better QoL as survivors. PMID- 16365609 TI - Gender disparities in percutaneous coronary interventions for acute myocardial infarction in Pennsylvania. AB - BACKGROUND: It has been shown that women are at greater risk than men of not receiving screening and treatment services for coronary heart disease. The purpose of this research was to determine whether there were gender disparities in the use of percutaneous coronary interventions (PCI) in the treatment of acute myocardial infarction (AMI) in Pennsylvania in 2000 and, if so, whether outcomes were affected. METHODS: Data included 10,170 patients treated with PCI and 21,181 patients medically managed in Pennsylvania hospitals. Multivariate analyses were performed using logistic regression to estimate the impact of gender on PCI. In addition, we performed retrospective matching on propensity scores to compare outcomes for women who were treated with PCI to comparable groups of women and men. RESULTS: After controlling for age, race/ethnicity, severity at admission, location of infarct, and source of admission, women had 24% lower odds than men of receiving PCI (P<0.0001). In a propensity score-matched sample of 3023 women who received PCI and 3023 women who did not, women who received PCI were significantly less likely to die (2.3% vs. 10.4%, P<0.0001). In a second propensity score-matched sample of 3329 women and 3329 similar men who received PCI, the difference in mortality was not statistically significant (1.59% vs. 1.92%, P=0.39). CONCLUSIONS: These results suggest that the morbidity and mortality associated with AMI in women could be reduced by increased use of PCI and that more women admitted for AMI should receive consideration for PCI. PMID- 16365610 TI - Disenrollment from an acute/long-term managed care program (PACE). AB - BACKGROUND: The Program of All-Inclusive Care for the Elderly (PACE) is an acute/long-term managed care plan designed to care for the most frail and vulnerable Medicare beneficiaries. To our knowledge, this is a first study to examine patterns and predictors of disenrollment from PACE. OBJECTIVE: PACE, with its comprehensive delivery system, dual capitation, and a focus on the most vulnerable population, may be expected to achieve low rates of exit and little selective dissenrollment. This study examines whether these goals have been accomplished. RESEARCH DESIGN: The study includes 30 PACE programs and 14,657 individuals enrolled in them. Individual-level records, obtained from an administrative database, contain information on sociodemographics, caregiver support, health status and disability, medical history, service utilization, and disenrollment. Program-level variables also were included. Cox proportional hazard models, with time-varying and time-invariant covariates, were employed to predict time to disenrollment. RESULTS: Our findings show a low level of disenrollment. We find no increase in disenrollment risk by age, functional or cognitive impairment, Medicaid eligibility, or diagnoses. Certain characteristics (eg, nursing treatments) appear to reduce the disenrollment hazard, whereas others (eg, hospital admissions, private pay status) significantly increase it. The risk of disenrollment also increases with longer nursing home stays, until 80 90 days, whereupon it begins to decline. CONCLUSIONS: Both enrollee and program level attributes predict program disenrollment. Programmatic and quality of care improvements may be needed to further minimize disenrollment, particularly in programs experiencing rates that are substantially greater than the average. PMID- 16365611 TI - Therapeutic goal attainment in patients with hypertension and dyslipidemia. AB - BACKGROUND: Recent guidelines emphasize the need to assess and treat overall risk for cardiovascular disease through the concomitant management of multiple risk factors. We sought to ascertain treatment patterns and attainment of therapeutic goals in patients with isolated and concomitant hypertension and dyslipidemia, both with and without diabetes mellitus (DM) and symptomatic cardiovascular disease. METHODS: Inception cohorts of more than 41,000 newly diagnosed hypertension and dyslipidemia patients from 6 medical centers of the south central Veterans Affairs health care system were evaluated. Treatment patterns and goal attainment for low-density lipoprotein cholesterol (LDL-C; Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults guidelines: <160, <130, or 10 M, probably due to ion-pairing. Spectra at [M'OH](T) < 10 M have been interpreted quantitatively by assuming that the integrated peak area of the 620 cm(-1) mode is linearly proportional to [Al(OH)(4)(-)] at constant a(w) and that the only significant equilibrium in these systems is the formation of a dimer that can be represented as (Al(OH)(4))(2)(2-)(aq), although it may exist in an oxo-bridged form such as [(HO)(3)Al-O-Al(OH)(3)](2). The (aquated) species Na(+), OH(-), Al(OH)(4)(-), the dimer, and their ion-pairs, were sufficient to interpret all the Raman observations. No evidence was found for various other species that have been claimed to exist in concentrated alkaline aluminate solutions. PMID- 16365652 TI - Synthesis, crystal structure, magnetic properties and theoretical studies on a one-dimensional polynuclear copper(II) complex [Cu2(mu1,3-SCN)2(mu'1,3 SCN)2(MPyO)2]n. AB - A new one-dimensional polynuclear copper(II) complex [Cu(2)(mu(1,3) SCN)(2)(mu'(1,3)-SCN)(2)(MPyO)(2)](n)(where MPyO = 4-methylpyridine N-oxide) has been synthesized and its crystal structure determined by X-ray crystallography. In the complex there exist two kinds of bridging coordination modes, namely, mu(1,3)-SCN(-) equatorial-equatorial (EE) bridging ligand and micro'(1,3)-SCN(-) equatorial-axial (EA) bridging ligand. Two micro(1,3)-SCN(-) EE bridging ligands coordinate two copper(II) ions in a binuclear unit, and the S atoms from the micro'(1,3)-SCN(-) EA bridging ligands as axial coordinated atoms link the binuclear units into one-dimensional chains. The ESR spectra have been investigated, and variable temperature (4-300 K) magnetic measurements were analyzed using a binuclear Cu(ii) magnetic interaction formula and indicate the existence of strong antiferromagnetic coupling with 2J=- 216.00 cm(-1) between bridged copper(II) ions. Density functional calculations have been carried out on this binuclear unit, yielding a similar singlet-triplet splitting. The mechanism of strong antiferromangetic interaction is revealed according to the calculations. PMID- 16365654 TI - Stabilization of the tautomers HP(OH)2 and P(OH)3 of hypophosphorous and phosphorous acids as ligands. AB - Treatment of [CpRu(PPh(3))(2)Cl] 1 with the stoichiometric amount of H(3)PO(2) or H(3)PO(3) in the presence of chloride scavengers (AgCF(3)SO(3) or TlPF(6)) yields compounds of formula [CpRu(PPh(3))(2)(HP(OH)(2))]Y (Y = CF(3)SO(3) 2a or PF(6) 2b) and [CpRu(PPh(3))(2)(P(OH)(3))]Y (Y = CF(3)SO(3) 3aor PF(6) 3b) which contain, respectively, the HP(OH)(2) and P(OH)(3) tautomers of hypophosphorous and phosphorous acids bound to ruthenium through the phosphorus atom. The triflate derivatives 2a and 3a react further with hypophosphorous or phosphorous acids to yield, respectively, the complexes [CpRu(PPh(3))(HP(OH)(2))(2)]CF(3)SO(3) 4 and [CpRu(PPh(3))(P(OH)(3))(2)]CF(3)SO(3) 5 which are formed by substitution of one molecule of the acid for a coordinated triphenylphosphine molecule. The compounds 2 and 3 are quite stable in the solid state and in solutions of common organic solvents, but the hexafluorophosphate derivatives undergo easy transformations in CH(2)Cl(2): the hypophosphorous acid complex 2b yields the compound [CpRu(PPh(3))(2)(HP(OH)(2))]PF(2)O(2) 6, whose difluorophosphate anion originates from hydrolysis of PF(6)(-); the phosphorous acid complex 3b yields the compound [CpRu(PPh(3))(2)(PF(OH)(2))]PF(2)O(2) 7, which is produced by hydrolysis of hexafluorophosphate and substitution of a fluorine for an OH group of the coordinated acid molecule. All the compounds have been characterized by elemental analyses and NMR measurements. The crystal structures of 2a, 3a and 7 have been determined by X-ray diffraction methods. PMID- 16365653 TI - Lanthanide complexes of chiral 3 + 3 macrocycles derived from (1R,2R)-1,2 diaminocyclohexane and 2,6-diformyl-4-methylphenol. AB - The enantiopure amine macrocycle H(3)L, as well as the parent macrocyclic Schiff base H(3)L1, the 3 + 3 condensation product of (1R,2R)-1,2-diaminocyclohexane and 2,6-diformyl-4-methylphenol, are able to form mononuclear complexes with lanthanide(III) ions. The lanthanide(III) complexes of H(3)L have been studied in solution using NMR spectroscopy and electrospray mass spectrometry. The NMR spectra indicate the presence of complexes of low C(1) and C(2) symmetry. The (1)H and (13)C NMR signals of the Lu(III) complex obtained from H(3)L have been assigned on the basis of COSY, TOCSY, NOESY, ROESY and HMQC spectra. The NMR data reveal unsymmetrical binding of lanthanide(III) ion and the presence of a dynamic process corresponding to rotation of Lu(III) within the macrocycle. The [Ln(H(4)L)(NO(3))(2)](NO(3))(2)(Ln = Sm(III), Eu(III), Dy(III), Yb(III) and Lu(III)) complexes of the cationic ligand H(4)L(+) have been isolated in pure form. The X-ray analysis of the [Eu(H(4)L)(NO(3))(2)](NO(3))(2) complex confirms the coordination mode of the macrocycle determined on the basis of NMR results. In this complex the europium(III) ion is bound to three phenolate oxygen atoms and two amine nitrogen atoms of the monoprotonated macrocycle H(4)L(+), as well as to two axial bidendate nitrate anions. In the presence of a base, mononuclear La(III), Ce(III) and Pr(III) complexes of the deprotonated form of the ligand L(3 ) can be obtained. When 2 equivalents of Pr(III) are used in this synthesis Na(3)[Pr(2)L(NO(3))(2)(OH)(2)](2)NO(3).5H(2)O is obtained. The NMR, ES MS and an X-ray crystal model of this complex show coordination of two Pr(III) ions by the macrocycle L. The X-ray crystal structure of the free macrocycle H(3)L1 has also been determined. In contrast to macrocyclic amine H(3)L, the Schiff base H(3)L1 adopts a cone-type conformation resembling calixarenes. PMID- 16365655 TI - Molybdenum and copper catalysis of reductions by titanium(II) and titanium(III). AB - Reductions of vanadium(IV), benzoquinone, and tri-iodide, both by titanium(III) and by titanium(II), are catalyzed by molybdenum(VI). The VO(2+)-Ti(II) reaction is catalyzed by copper(II) as well. Reactions of Ti(II) with the oxidant in excess yield Ti(IV), as do reductions by Ti(III). Reactions proceed via competing uncatalyzed and catalyzed paths, with the latter components first order in catalyst. Kinetic patterns indicate that monomeric Mo(v) is the active species, but the dimeric Mo(v) species, [Mo(2)O(4)](2+), is without catalytic action. Catalytic constants pertaining to Ti(III) are remarkably similar to those for Ti(II), despite the 0.47 V difference in the standard potentials of the two reductants. PMID- 16365657 TI - Editorial. PMID- 16365656 TI - Synthetic and reaction chemistry of heteroatom stabilized boryl and cationic borylene complexes. AB - The synthesis, spectroscopic and structural characterization of the aryloxy and amino functionalized chloroboryl complexes (eta(5)-C(5)R(5))Fe(CO)(2)B(OMes)Cl (R = H, 2a ; R = Me, 3a) and (eta(5)-C(5)H(5))Fe(CO)(2)B(N(i)Pr(2))Cl (7a) are reported. Compound 2ais shown to be a versatile substrate for further boron centred substitution chemistry leading to the asymmetric boryl complexes (eta(5) C(5)H(5))Fe(CO)(2)B(OMes)ER(n) [ER(n) = OC(6)H(4)(t)Bu-4, 2c; ER(n) = SPh, 2d] with retention of the metal-boron bond. The reactivities of 2a, 3a and 7a towards the halide abstraction agent Na[BAr(f)(4)] have also been examined, in order to investigate the potential for the generation of cationic heteroatom-stabilized terminal borylene complexes. The application of this methodology to the mesityloxy derivatives and gives rise to B-F containing products, presumably via fluoride abstraction from the [BAr(f)(4)](-) counter-ion. By contrast, amino functionalized complex 7a is more amenable to this approach, and the thermally robust terminal aminoborylene complex [(eta(5) C(5)H(5))Fe(CO)(2)B(N(i)Pr(2))][BAr(f)(4)] (9) can be isolated in ca. 50% yield. The reactivity of 9towards a range of nucleophilic and/or unsaturated reagents has been examined, with examples of addition, protonolysis and metathesis chemistries having been established. PMID- 16365658 TI - Jane Thomas-Oates, University of York. AB - The Analyst profiles Jane Thomas-Oates, RSC/EPSRC Chair of Analytical Science at the University of York, UK. PMID- 16365659 TI - Advances and challenges in the identification of volatiles that mediate interactions among plants and arthropods. AB - The relatively new research field of Chemical Ecology has, over the last two decades, revealed an important role of plant-produced volatile organic compounds (VOCs) in mediating interactions between plants and other organisms. Of particular interest are the volatile blends that plants actively emit in response to herbivore damage. Various efforts are underway to pinpoint the bioactive compounds in these complex blends, but this has proven to be exceedingly difficult. Here we give a short overview on the role of herbivore-induced plant volatiles in interactions between plants and other organisms and we review methods that are currently employed to collect and identify key volatile compounds mediating these interactions. Our perspective on future directions of this fascinating research field places special emphasis on the need for an interdisciplinary approach. Joint efforts by chemists and biologists should not only facilitate the elucidation of crucial compounds, but can also be expected to lead to an exploitation of this knowledge, whereby ecological interactions may be chemically manipulated in order to protect crops and the environment. PMID- 16365660 TI - Investigation of selenium-containing root exudates of Brassica juncea using HPLC ICP-MS and ESI-qTOF-MS. AB - Selenium-containing root exudates were investigated in a known selenium accumulator model plant. Indian mustard (Brassica juncea) plants were grown hydroponically and supplemented with selenite (SeO(3)(2-)) in a 25% Hoagland's nutrient solution. Additive concentrations were 0, 1, 5 and 20 microg mL(-1) Se with five replicate plants per treatment level. Plants were exposed to the respective Se solutions for two weeks, then placed in deionized water for two more weeks. The hydroponic solutions were collected for analysis after the first two weeks of selenium supplementation (day 14) and twice during the deionized water period (days 21 and 28). Separation by ion-pairing high performance liquid chromatography was followed by inductively coupled plasma-mass spectrometry (ICP MS) for selenium specific detection. Chromatographic peaks unable to be identified by retention-time matching were collected for analysis by electrospray ionization mass spectrometry (ESI-MS). Additional chemical experiments were performed for structural elucidation. Several selenium-containing compounds were identified in the exudate-containing solution and two were identified as selenocystine and the selenosulfate (SSeO(3)(2-)) ion. The presence of dimethylselenide (CH(3)SeCH(3)) is also observed but cannot be attributed exclusively to plant exudation because plants were not grown in sterile conditions. Further, the incorporation of fortified selenoamino acids into peptide structures was found to occur under neutral pH conditions, suggesting that exuded enzymes might facilitate such a reaction. Finally, physiological differences resulting from selenium supplementations were noted and discussed. PMID- 16365661 TI - Incorporation of a DNAzyme into Au-coated nanocapillary array membranes with an internal standard for Pb(ii) sensing. AB - A Pb(ii)-specific DNAzyme has been successfully incorporated into Au-coated polycarbonate track-etched (PCTE) nanocapillary array membranes (NCAMs) by thiol gold immobilization. Incorporation of the DNAzyme into the membrane provides a substrate-bound sensor using a novel internal control methodology for fluorescence-based detection of Pb(ii). A non-cleavable substrate strand, identical to the cleavable DNAzyme substrate strand except the RNA-base is replaced by the corresponding DNA-base, is used for ratiometric comparison of intensities. The cleavable substrate strand is labeled with fluorescein, and the non-cleavable strand is labeled with a red fluorophore (Cy5 or Alexa 546) for detection after release from the membrane surface. This internal standard based ratiometric method allows for real-time monitoring of Pb(ii)-induced cleavage, as well as standardizing variations in substrate size, solution detection volume, and monolayer density. The result is a Pb(ii)-sensing structure that can be stored in a prepared state for 30 days, regenerated after reaction, and detect Pb(ii) concentrations as low as 17 nM (3.5 ppb). PMID- 16365662 TI - Planar nitric oxide (NO)-selective ultramicroelectrode sensor for measuring localized NO surface concentrations at xerogel microarrays. AB - A planar ultramicroelectrode nitric oxide (NO) sensor was fabricated to measure the local NO surface concentrations from NO-releasing microarrays of varying geometries. The sensor consisted of platinized Pt (25 microm) working electrode and a silver paint reference electrode coated with a thin silicone rubber gas permeable membrane. An internal hydrogel layer separated the Pt working electrode and gas permeable membrane. The total diameter of the sensor was 3.5 provokes a strong rebind of the template in both imprinted and non-imprinted polymers. This pH dependency can be explained as a selective chemical modification which reduces bind diversity following a model based on enolization by protonation of the specific cavities. Characterization of the adsorption isotherms showed good agreement with the Langmuir-Freundlich (LF) model, presenting quite homogeneous binding sites for a bulk material and high capacity in the imprinting pH range. In addition, the affinity spectrum (AS) method has been represented showing the typical profiles of LF isotherm for both sub-saturation and saturation levels, being in general agreement with the encountered values for fitting coefficients. The covalent molecular imprinted polymer has been successfully evaluated in a SPE process for further OTC determination in the certified mussel tissue (CRM 477). PMID- 16365670 TI - The electroanalytical detection of hydrazine: a comparison of the use of palladium nanoparticles supported on boron-doped diamond and palladium plated BDD microdisc array. AB - We show that both a random distribution of palladium nanoparticles supported on a BDD electrode or a palladium plated BDD microelectrode array can each provide a sensing platform for the electrocatalytic detection of hydrazine. The palladium nanoparticle modified electrode displays a sensitivity and limit of detection of 60 mA mol(-1) L and 2.6 microM respectively while the array has a sensitivity of 8 mA mol(-1) L with a detection limit of 1.8 microM. The beneficial cost implications of using palladium nano- or micro-particles in sensors compared to a palladium macroelectrode are evident. Interestingly the array of the nanoparticles shows similar sensitivity and limit of detection to the microelectrode array which probably indicates that the random distribution of the former leads to 'clumps' of nanoparticles that effectively act as microelectrodes. PMID- 16365671 TI - Measurement strategy and instrumental performance of a computer screen photo assisted technique for the evaluation of a multi-parameter colorimetric test strip. AB - A measuring strategy for the evaluation of a seven parameters colorimetric test using a computer screen photo-assisted technique (CSPT) is demonstrated. CSPT is a versatile approach aimed at point of care or home tests that uses regular computer sets and web cameras as the whole instrument. Issues such as the stability and the equivalency on different platforms of the determinations have been addressed in the present work. The method uses an embedded local reference simultaneously measured with the tests and solves the evaluation as a classification problem. The achieved performance tested along 580 classifications covering all the ranges of the assay, using synthetic samples, yielded 97.2% correct determinations compared with 89.7% for the case of colorimetric determinations. The errors were concentrated in only two parameters that show a significant correlation with a set of quality indices used to assess the performance of the classification. PMID- 16365672 TI - Effect of fingerprint conformation and spectral scaling on the performance of computer screen photo-assisted experiments. AB - The use of computer screens as controlled light sources and web cameras as image detectors (the so-called computer screen photo-assisted technique, CSPT) is an ubiquitous alternative for the evaluation of colorimetric quick tests at homes or in primary care units. The performance of CSPT for such evaluations depends on several factors, from which the most relevant are the composition of illuminating sequences and the conformation of CSPT substance signatures. In this work, with the aid of a CSPT model, the effect of the construction of the substance signatures on the classification performance of different representative substance sets is studied. The correlation of illuminating colors with such classification is investigated, allowing one to determine redundancy and limitations with respect to visible spectroscopy. The concept of spectral scaling is introduced and its properties compared with standard procedures. PMID- 16365673 TI - Detection of trace materials with Fourier transform infrared spectroscopy using a multi-channel detector. AB - FTIR spectroscopy is one of the most powerful methods for material characterization. However, the sensitivity of this analytical tool is often very limited especially for materials with weak infrared absorption or when spectral bands of the targeted trace material overlap with the spectral bands of major components. Fortunately, for heterogeneous samples, there is an opportunity to improve the sensitivity of detection by using an imaging approach. This paper explores the opportunity of enhancing the sensitivity of FTIR spectroscopy to detect trace amounts of materials using the FTIR imaging approach based on a focal plane array (FPA) detector. Model sample tablets of ibuprofen in hydroxypropyl methylcellulose (HPMC) have been used to exemplify the detection limits of FTIR spectroscopy using: (a) a conventional mercury cadmium telluride (MCT) detector and (b) a FPA detector. The sensitivity level was compared and it has been found that for this particular set of samples, the lowest concentration of ibuprofen in HPMC that can be detected using attenuated total reflection (ATR) measuring mode with the single element MCT detector was 0.35 wt% while using the FPA detector, the presence of drug has been detected in a sample that contains as little as 0.075 wt% of drug. The application of using this enhanced sensitivity offered by the multi-channel detector to probe trace amounts of drug particles left on the surface of a finger after handling a small amount of the drug has also been demonstrated. These results have broad implications for forensic, biomedical and pharmaceutical research. PMID- 16365674 TI - On-line dynamic extraction and automated determination of readily bioavailable hexavalent chromium in solid substrates using micro-sequential injection bead injection lab-on-valve hyphenated with electrothermal atomic absorption spectrometry. AB - A novel and miniaturized micro-sequential injection bead-injection lab-on-valve (microSI-BI-LOV) fractionation system was developed for on-line microcolumn soil extraction under simulated environmental scenarios and accurate monitoring of the content of easily mobilisable hexavalent chromium in soil environments at the sub low parts-per-million level. The flow system integrates dynamic leaching of hexavalent chromium using deionized water as recommended by the German Standard DIN 38414-S4 method; on-line pH adjustment of the extract by a 0.01 mol L(-1) Tris-HNO(3) buffer solution; isolation of the chromate leached from the matrix constituents onto a Q Sepharose strong anion-exchanger freshly packed into the microconduits of the microSI-assembly; air-segmented elution of the sorbed species by a 40 microL plug of 0.5 mol L(-1) NH(4)NO(3) (pH 8) eluent; and detection by electrothermal atomic absorption spectrometry (ETAAS). The effect of simulated acidic rain on the accessibility of chromate forms for plant uptake was also investigated. The proposed approach offers several advantages over conventional speciation/fractionation protocols in the batch mode, including immediate separation with concomitant preconcentration of the released chromate, minimization of Cr(vi) to Cr(iii) interconversion risks, enhanced accuracy, and non-existence of re-adsorption/re-distribution problems along with a detailed pattern of the kinetics of the leaching process. The reliability of the proposed method was evaluated via spiking of a moderately polluted agricultural soil material (San Joaquin Soil-Baseline Trace Element Concentrations) with water soluble Cr(vi) salts at different concentration levels. The potential of the microSI-BI-LOV set-up with renewable surfaces for flame-AAS determination of high levels of readily bioavailable chromate in contaminated soils is also addressed. PMID- 16365676 TI - Distinction of gases with a semiconductor sensor depending on the scanning profile of a cyclic temperature. AB - A gas-sensing system based on a dynamic nonlinear response is reported to improve the selectivity in the sensor response toward sample gases. A cyclic temperature composed of fundamental and second harmonics was applied to a SnO(2) semiconductor gas sensor and the resulting conductance of the sensor was analyzed by fast Fourier transformation (FFT). The dynamic nonlinear responses to the gas species were further characterized depending on the scanning profile of the temperature. These characteristic sensor responses under the application of second-harmonic perturbation were theoretically considered based on a reaction diffusion model for the semiconductor surface. PMID- 16365675 TI - Flow injection on-line solid phase extraction for ultra-trace lead screening with hydride generation atomic fluorescence spectrometry. AB - A flow injection (FI) on-line solid phase extraction (SPE) procedure for ultra trace lead separation and preconcentration was developed, followed by hydride generation and atomic fluorescence spectrometric (AFS) detection. Lead is retained on an iminodiacetate chelating resin packed microcolumn, and is afterward eluted with 2.5% (v/v) hydrochloric acid to facilitate the hydride generation by reaction with alkaline tetrahydroborate solution with 1% (m/v) potassium ferricyanide as an oxidizing (or sensitizing) reagent. The hydride was separated from the reaction medium in the gas-liquid separator and swept into the atomizer for quantification. The chemical variables and the FI flow parameters were carefully optimized. With a sample loading volume of 4.8 ml, quantitative retention of lead was obtained, along with an enrichment factor of 11.3 and a sampling frequency of 50 h(-1). A detection limit of 4 ng l(-1), defined as 3 times the blank standard deviation (3 sigma), was achieved along with a RSD value of 1.6% at the 0.4 microg l(-1) level. The procedure was validated by determining lead contents in two certified reference materials, and its practical applicability was further demonstrated by analysing a variety of biological and environmental samples. PMID- 16365677 TI - Piezoelectric immunosensor for bisphenol A based on signal enhancing step with 2 methacrolyloxyethyl phosphorylcholine polymeric nanoparticle. AB - An immunoassay in which BPA competed with a BPA-horseradish peroxidase conjugate for binding to anti-BPA antibodies, coupled to a piezoelectric (PZ) immunosensor, was able to detect 0.1 ng mL(-1) BPA. To enhance the sensitivity of the assay, we tested nanoparticles approximately 200 nm in diameter, coupled to anti-BPA antibodies, to increase the mass change on the surface of the immunosensor and thereby increase the frequency shift detected. This second step, using nanoparticles coated with anti-BPA antibodies, improved the sensitivity of the assay by approximately eight times at BPA concentrations below 10 ng mL(-1). Field emission-scanning electron microscopy (FE-SEM) showed that polymeric 2 methacrolyloxyethyl phosphorylcholine (MPC) nanoparticles coupled to antibodies remained monodisperse on the surface of the immunosensor and therefore produced stable signals in the immunosensors. Since the frequency shift detected in the assay mainly originated from the mass change on the surface of the PZ crystal, the colloidal stability of the antibody-conjugated particles used in the enhancement step played an extremely important role in achieving a stable and highly sensitive signal. PMID- 16365678 TI - Over-expression of tobacco NtHSP70-1 contributes to drought-stress tolerance in plants. AB - HSP70, a heat shock protein, is a molecular chaperone responsive to various environmental stresses. Here, NtHSP70-1 was a drought-/ABA-inducible gene. We monitored the expression of CaERD15 (early responsive to dehydration) with exposing plants to progressive drought stress. Its activity was used as an indicator of water-deficit conditions. To analyze the protective role of HSP70, we obtained transgenic tobacco plants that constitutively expressed elevated levels of the tobacco HSP70, NtHSP70-1, as well as transgenic plants containing either the vector alone or else having NtHSP70-1 in the antisense orientation. Plants with enhanced levels of NtHSP70-1 in their transgenic sense lines exhibited tolerance to water stress. Under progressive drought, the amount of leaf NtHSP70-1 was correlated with maintenance of optimum water content, with contents being higher in the leaves of dehydrated transgenic sense plants than in those of either the control (vector-only) or the transgenic antisense plants. Moreover, the expression of CaERD15 was considerably reduced in tobacco plants that over-expressed NtHSP70-1. These results suggest that elevated levels of NtHSP70-1 can confer drought-stress tolerance. PMID- 16365679 TI - Induction of a cytosolic pyruvate kinase 1 gene during the resistance response to Tobacco mosaic virus in Capsicum annuum. AB - Hot pepper (Capsicum annuum L. cv. Bugang) plants exhibit a hypersensitive response (HR) upon infection by Tobacco mosaic virus (TMV) pathotype P(0). Previously, to elucidate molecular mechanism that underlies this resistance, hot pepper cv. Bugang leaves were inoculated with TMV-P(0) and genes specifically up regulated during the HR were isolated by microarray analysis. One of the clones, Capsicum annuum cytosolic pyruvate kinase 1 (CaPK(c)1) gene was increased specifically in the incompatible interaction with TMV-P(0). The expression of CaPK(c)1 gene was also triggered not only by various hormones such as salicylic acid (SA), ethylene, and methyl jasmonate (MeJA), but also NaCl and wounding. These results suggest that CaPK(c)1 responds to several defense-related abiotic stresses in addition to TMV infection. PMID- 16365680 TI - Adenocarcinoma of the ileoanal pouch for ulcerative colitis--a complication of severe chronic atrophic pouchitis? AB - BACKGROUND: The appearance of a carcinoma in the ileal pouch after restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) for ulcerative proctocolitis is rare. Most of these adenocarcinomas previously described in literature develop from residual viable rectal mucosa. We report a case of an adenocarcinoma arising in all probability from the ileal pouch after malignant transformation of the ileal pouch mucosa based on a chronic atrophic pouchitis. PATIENT AND METHODS: A 34-year-old man developed an adenocarcinoma after a double stapled ileorectal J-pouch for ulcerative colitis (UC) proceeded from malignant ileal transformation. Before surgery, he had a 20-year history of UC refractory to medical therapy, but no occurrence of backwash ileitis, dysplasia or colitis associated illness. He experienced severe pouchitis after IPAA since the ileostomy closure. Carcinoma was ensured by endoscopy, and the patient underwent an abdominoperineal pouch extirpation combined with excision of perirectal tissues and anal canal. Histology after surgery showed a pT4,pN2(4/16)pM0,G3 adenocarcinoma with global severe chronic atrophic pouchitis (CAP), villous atrophy and malignant ileal transformation. No metaplasia of the rectal mucosa was found, not even malignant epithelial transformation of the anal canal. CONCLUSION: This case suggests that a malignant transformation of the ileal pouch mucosa may occur as a pure complication of severe CAP, even in the absence of backwash ileitis or a previous history of cancer. The absence of metaplasia of the rectal mucosa revealed the passage from CAP to dysplastic epithelium and to cancer. A multifactorial development of carcinogenesis is supposed, but we emphasize the importance of severe CAP, and that careful surveillance is needed in patients after IPAA. We must submit that this is just a case report and cannot stand for general cancer development in ulcerative colitis, but it may point out the risk factor of chronic inflammation and leads the surgeon to skillful working when building the pouch. PMID- 16365681 TI - Androgen receptor CAG and GGC polymorphisms in Mediterraneans: repeat dynamics and population relationships. AB - Microsatellite variation (CAG and GGC repeats) of the androgen receptor (AR) gene shows remarkable differences among African and non-African populations. In vitro studies have demonstrated an inverse relationship between the length of both microsatellites and AR activity. This fact may explain the observed association of the AR gene with prostate cancer and the strong ethnic differences in the incidence of this cancer. CAG and GGC genetic variation has been tested in a large set of populations from the Ivory Coast as well as 12 Mediterranean samples whose variation is described for the first time. The pattern of frequencies observed in the Ivory Coast agrees with data previously reported for other Sub Saharan populations. Concerning the Mediterranean variation, Sardinian samples are characterised by low genetic diversities, and Egyptian Siwa Berbers by a particular pattern of GGC frequencies. High and Middle Atlas Moroccan Berbers are the most closely related to the Sub-Saharan variation. For both the CAG and GGC repeats, the Ivory Coast and some Moroccan samples exhibit high frequencies of low size alleles (CAG under 18 repeats, and GGC under 15 repeats) that have been associated with prostate cancer. PMID- 16365682 TI - Phenotypic and molecular characterisation of a de novo 5q deletion that includes the APC gene. AB - We report on a 12-year-old female patient with mild dysmorphic signs, including bilateral epicanthal folds, low-set dysplastic ears, a short nose with anteverted nostrils, conically shaped fingers, generalised increase of subcutaneous fat, multiple fine venous teleangiectasia on her back, mild pectus carinatum, and a general muscular hypotonia. Cytogenetic analysis and fluorescence in situ hybridisation (FISH) studies using region-specific BAC and YAC clones indicated a de novo interstitial deletion of the long arm of chromosome 5, resulting in monosomy 5q21.1-q23.1. Molecular analysis of polymorphic markers helped to narrow down the breakpoints and demonstrated that the derivative chromosome 5 is of paternal origin. By using the same panel of polymorphic markers, a reinvestigation of a similar, already published, 5q deletion case [Raedle et al. (2001) Am J Gastroenterol 96:3016-3020] was performed, allowing a more detailed genotype-phenotype correlation. Phenotypic classification was also carried out. Several known genes, including APC and MCC, were found to map to the common deleted genomic segment. Genetic counselling based on the molecular analysis data was performed for the index family. PMID- 16365683 TI - ALDH2 and CYP2E1 genotypes, urinary acetaldehyde excretion and the health consequences in moderate alcohol consumers. AB - Deficiencies in mitochondrial low-Km aldehyde dehydrogenase (ALDH2) activity, and consequently high blood acetaldehyde levels, have been suggested to relate to various diseases in Japanese, including esophageal cancer. In the present study, 200 men aged 35-59 years randomly selected from an occupational population were analyzed for the association of ALDH2 genotypes and cytochrome P450-2E1 (CYP2E1) genotypes with the urinary excretion of acetaldehyde (which is bound to some chemicals in the urine) and with common alcohol-related health consequences. Urinary acetaldehyde excretion was increased, reflecting increased alcohol consumption even in this moderate alcohol-consuming population. Neither the ALDH2 nor the CYP2E1 genotypes showed significant influence on the elevation of urinary acetaldehyde excretion. Neither these genotypes nor urinary acetaldehyde concentration significantly affected blood pressure, serum aspartate aminotransferase and gamma-glutamyl transferase activities, or serum HDL cholesterol and lipid peroxide concentrations. It was concluded that acetaldehyde accumulates in moderate alcohol consumers irrespective of ALDH2 and CYP2E1 genotype, and that the implications of these genotypes and acetaldehyde accumulation in terms of common alcohol-related health consequences were obscure. The results also suggest that the carcinogenicity of acetaldehyde on esophageal mucosa depends greatly upon repeated exposure to high blood acetaldehyde, even through transient rather than chronic exposure. PMID- 16365684 TI - CPPD crystal deposition disease in patients with rheumatoid arthritis. AB - The aim of this study was to assess the frequency and the outcome of patients suffering from rheumatoid arthritis in which calcium pyrophosphate dihydrate (CPPD) crystal deposits were found to coexist in synovial fluid analysis. Such association was more frequent than previously believed with CPPD crystals found in 25.8% of 93 patients with rheumatoid arthritis. As a group, a trend toward a worse outcome was suggested by more frequent prostheses of the lower limb. PMID- 16365685 TI - Disease activity and functional changes of RA patients receiving different DMARDs in clinical practice. AB - The aim of this study was to compare the effectiveness of different disease modifying anti-rheumatic drugs (DMARDs) in improving disease activity and functional status in patients with rheumatoid arthritis (RA). One hundred and fifty-two Thai RA patients treated with at least one DMARD were enrolled in this 1-year cohort. Demographic characteristics, baseline and end-of-study data on disease activity and Health Assessment Questionnaire (HAQ) Disability Index of the subjects were compared among different DMARD options. Predictors of HAQ score were investigated using regression analysis. The results showed that the studied patients had established RA with mild to moderate activity. More than 85% were prescribed methotrexate (MTX) as single or combined DMARDs. At 1 year, improvement in most activity measures was experienced. However, all patients had functional declines. Patients taking antimalarial agents had the maximal rate of functional deterioration. Patients taking MTX-based DMARDs had significantly lower rate of functional decline than patients taking DMARDs without MTX (p=0.018). Only patients receiving MTX-based DMARDs had clinically meaningful improvement in HAQ score. The predictors of HAQ score at 1 year included baseline HAQ score and patient global assessment at end of study. In conclusion, although DMARD treatment was shown to improve disease activity in RA patients, functional deterioration was evident in our cohort. Thus, measures of functional status are more appropriate than measures of disease activity to evaluate treatment effects of DMARDs in established RA in clinical practice. MTX-based DMARDs should be prescribed where possible in RA patients with persisting activity due to their ability to delay functional deterioration. PMID- 16365686 TI - Is there a hypothalamic-pituitary-adrenal axis dysfunction in patients with familial Mediterranean fever? PMID- 16365687 TI - Early assessment of rapidly progressive interstitial pneumonia associated with amyopathic dermatomyositis. AB - Amyopathic dermatomyositis (ADM) is occasionally complicated by rapidly progressive interstitial pneumonia (RPIP), and in such cases, diffuse alveolar damage (DAD) is usually diagnosed at autopsy. Here, we present three patients with RPIP accompanied by ADM in whom lung disease was assessed at an early stage. High-resolution computed tomography (HRCT) carried out before the onset of dyspnoea revealed uniformly subpleural reticular opacity with faint ground-glass attenuation. At that stage, surgical lung biopsies from two patients showed histological patterns typical of cellular nonspecific interstitial pneumonia (NSIP). Despite pulse methylprednisolone and subsequent high-dose oral administration of prednisolone, lung disease progressed in all patients, with extensive areas of ground-glass opacity and consolidation observed in HRCT scans. DAD was confirmed histologically in one case. Additional administration of cyclosporine, pulse cyclophosphamide or high-dose intravenous administration of immunoglobulin rescued all patients. Our data suggest that ADM-associated interstitial pneumonia takes an aggressive course even when the radiological and histological features are consistent with NSIP. Aggressive combination therapy with high-dose steroids and immunosuppressive agents is required as early as possible for patients with this life-threatening disorder. PMID- 16365688 TI - Human foamy virus bel1 sequence in patients with autoimmune rheumatic diseases. AB - Since the association between human foamy virus (HFV) with rheumatic autoimmune diseases remains controversial, this study was designed to determine the relationship between HFV and systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), or progressive systemic sclerosis (PSS). The bel1 and Pol sequences of HFV were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) in plasma and by PCR in peripheral blood mononuclear cells (PBMC) from patients with SLE, RA, and PSS. Antibodies against Bel1 and Pol were assessed by enzyme-linked immunosorbent assay. Active HFV infections were detected by a Bel1 responsive indicator cell line. The bel1 sequence was detected in the plasma (SLE 59, RA 32, and PSS 63%) and PBMC (SLE 54, RA 71, and PSS 57%). However, active HFV infection existed only in patients with the bel1 sequence in both plasma and PBMC. In SLE patients, antibodies against Bel1 (7.1%) and Pol (4.5%) were also detected. The results suggest a possible association between HFV infection and these autoimmune rheumatic diseases. PMID- 16365689 TI - A case of systemic lupus erythematosus complicated by alveolar hemorrhage and cytomegalovirus colitis. AB - We report a rare case of systemic lupus erythematosus (SLE) complicated by alveolar hemorrhage and cytomegalovirus (CMV) colitis. Despite the successful treatment of lupus nephritis by steroid pulse therapy, the patient developed an acute alveolar hemorrhage 2 months later. Cyclophosphamide pulse therapy ameliorated the hemorrhage. One month later, she suddenly developed melena secondary to CMV colitis. Antiviral therapy was successful. We emphasize the importance of timely and precise differential diagnosis for successful management of complicated SLE. PMID- 16365690 TI - A multicenter study of patients with adult-onset Still's disease compared with systemic juvenile idiopathic arthritis. AB - Adult-onset Still's disease (AOSD) has often been regarded as the adult spectrum of systemic juvenile idiopathic arthritis (sJIA). The present study aims to compare the clinical and laboratory features, the disease course and the response to treatment in patients having AOSD with those having sJIA. Retrospective review of all available data that were filled out by adult and paediatric rheumatologists from six centers using a standard data extraction form was performed. A total of 95 patients with AOSD and 25 patients with sJIA were recruited for the study. The frequency of fever, rash, myalgia, weight loss and sore throat was higher in patients with AOSD. The pattern of joint involvement differed slightly. Laboratory findings were similar in both groups, except that liver dysfunction and neutrophilia were more common among adults. A multiphasic pattern dominated the childhood cases, whereas the most frequent course was a chronic one in adults. Corticosteroids and methotrexate were the most commonly employed therapy; however, chloroquine was another popular therapy in the adult group. We showed a difference in the rate of clinical and laboratory features between patients with AOSD and those with sJIA. AOSD and sJIA may still be the same disease, and children may simply be reacting differently as the result of the first encounter of the putative antigens with the immune system. PMID- 16365691 TI - Toxic effects of povidone-iodine on synovial cell and articular cartilage. AB - The objective of this study was to investigate the toxic effects of povidone iodine (PVI). A series of MTT assay of cultured synovial cells stimulated by PVI have been made. Test for assay of membrane-toxic effect of PVI and lactate dehydrogenase (LDH) has also been carried out. After making intraarticular injection into rats' hind knee joints with 0.1 ml of PVI every day for 1 week, we have extracted the knee joints and blood from rats in four divided groups on the 0, 1st, 3rd, and 7th day. Our microscopic analysis was performed that cartilage and synovium membrane were stained with hematoxylin, toluidine blue, and safranin O. Iodine, glutamic oxaloacetic transminase, creatine phosphokinase (CPK), and LDH in rat sera have been measured. We were able to confirm cytotoxicity in the synovial cells placed in PVI concentration of more than 6.25 x 10(-2)%. In comparison with the positive control, LDH, in this case, was released at the concentration level of 33.3%. Independent of period of stimulation, LDH was released the most in the PVI concentration of 6.25 x 10(-2). The histopathological analysis of articular cartilage has indicated clear morphological alterations in rats' synovium; especially, toluidine blue staining and safranin O staining were seen in the metachromasia of articular cartilage. However, no fissures or erosive changes in the articular cartilage, which were usually observable in the case of osteoarthritis, were observed. In the synovium inflammatory cells, which were mainly plasma cells and lymphoid cells, no neutrophils were infiltrated. The degree of infiltration has increased steadily day by day. Fibrin deposition has been observed even in the synovium of 1-day stimulation. On the 7th day, a serum total iodine concentration has been 544 microg/dl, 4,440 IU/ml for CPK concentration, and 2,049 IU/ml for LDH concentration, while the control concentrations have been 7.1 microg/dl, 491 IU/ml, and 700 IU/ml, respectively. Our experiment has disclosed that PVI preparation is strongly cytotoxic to synovial cell and articular cartilage. PMID- 16365692 TI - Retroperitoneal fibrosis and c-ANCA positivity. AB - Retroperitoneal fibrosis (RPF) is an uncommon collagen vascular disease characterized by a chronic nonspecific inflammation of the retroperitoneum, which can entrap and obstruct retroperitoneal structures. Although obscure, an autoimmune-mediated or vasculitic etiology has been hypothesized. In the current report, a case of RPF associated with the presence of antibodies against proteinase III, which responded effectively to immunosuppressive therapy, is presented. PMID- 16365693 TI - Familial transmission of genitovaginal prolapse. AB - Some females with little to no risk factors develop prolapse, while other females with multiple risk factors do not. It appears that some women may have a predisposition for prolapse in the setting of equivalent risk factors. We identified 10 patients younger than 55 years old with a family history of prolapse. Their average age was 37 years (range 27-51), the mean number of deliveries was 1.8, and their mean birth weight was 8 lbs. Genetic analysis of the inheritance pattern within these families demonstrated that pelvic organ prolapse segregated in a dominant fashion with incomplete penetrance in these families. Both maternal and paternal transmissions were observed. The relative risk to siblings of affected patients was five times that of the risk for the general population. Further investigation of these families may identify a genetic defect responsible for prolapse. PMID- 16365694 TI - Sensitivity and specificity of a new MRI method evaluating temporo-mandibular joint disc-condyle relationships: an in vivo study. AB - The aim of this study was to evaluate sensitivity and specificity of a new method to locate temporo-mandibular joint (TMJ) disc using magnetic resonance imaging (MRI) and analyze disc-condyle relationships, in asymptomatic subjects and patients with disc displacement. Twenty-nine sagittal MRI of 16 subjects, 8 asymptomatic volunteers and 8 subjects with anterior disc displacement, were carried out during controlled opening from intercuspal position up to a 25 mm opening. Selected sections were analyzed with a graphic computerized system of coordinates. The total surface area (TS) of disc section was separated into anterior surface area (AS) and posterior surface area. Areas were determined by computer. Two trained examiners drew images at random. The reliability of AS/TS ratio index was evaluated in a previous study. AS/TS ratio sensitivity (Se) and specificity (Sp) were calculated closed mouth, 5 mm open and 25 mm open mouth. Best sensitivity (Se=0.63) and specificity (Sp=0.81) were obtained when MRI was realized with closed mouth and 25 mm open mouth. Lower sensitivity was observed when MRI was performed either with closed mouth (Se=0.54) or 25 mm open mouth (Se=0.18). Lower specificity was observed with 5 mm open mouth (Sp=0.68). In conclusion, it was confirmed as well that MRI of anterior disc displacement should be performed with closed mouth and opened mouth. Thus, further studies are required to assess disc displacement and mechanical alterations and to evaluate the risk of direct damage on TMJ tissues. PMID- 16365695 TI - Branching patterns of the arterial branches supplying the middle vascular pedicle of the sternocleidomastoid muscle: a topographic anatomical study with surgical applications for the use of pedicles osteomuscular flaps. AB - When making a sternocleidomastoid (SCM) osteomuscular flap to include the clavicle and determining the rotation arc of the osteomuscular flap, it is very important to know the location and the origin of the superior thyroid artery and the distribution pattern of the SCM branch. Accordingly, in this study, the 50 SCM muscles and their arteries were dissected in 26 Korean cadavers, and the results were analyzed. The average distances from the origin of the superior thyroid artery to the clavicular and sternal heads of the SCM muscle were 87.6 mm (57.7-123.8 mm) and 131.2 mm (99.7-166.8 mm), respectively. The average distance from the origin of the superior thyroid artery to the SCM branch entering the SCM muscle was 30.1 mm (16.0-37.7 mm). After entering the SCM muscle, the SCM branches of the superior thyroid artery bifurcated into the clavicular and sternal branches at a point located an average of 58.8 mm (28.4-130.4 mm) above the clavicle. The distribution patterns of the superior thyroid artery were classified into six types based on the branching order and the dual supplies to the SCM muscle. Among them, type I in which the laryngeal branch first divided from the superior thyroid artery was the most common case (36%). PMID- 16365696 TI - Response of blood cell antioxidant enzyme defences to antioxidant diet supplementation and to intense exercise. AB - BACKGROUND: Exhaustive exercise induces oxidative stress. The cellular antioxidant defence systems have demonstrated great adaptation to chronic exercise. AIM: To establish the influence of the antioxidant diet supplementation on the erythrocyte and lymphocyte antioxidant enzyme activities in athletes at basal and post-exercise levels. METHODS: Fifteen amateur trained male athletes were randomly distributed in two groups: control and antioxidant supplemented (90 days' diet supplementation with 500 mg/day vitamin E and 30 mg/day beta-carotene, and the last 15 days also with 1 g/day vitamin C). The study was double blind. Maximal and submaximal exercise tests were performed after three months of diet supplementation. The study was developed during the training and competition season. RESULTS: The sportsmen of the supplemented group presented significantly higher plasmatic final levels of vitamin C, vitamin E and beta-carotene. Erythrocyte glutathione reductase activity significantly decreased in the placebo group but was maintained in the supplemented group after the three months studied. The erythrocyte superoxide dismutase activity increased after the training/competition period in the placebo group. Lymphocyte catalase and glutathione peroxidase activities increased significantly in the supplemented group after the supplementation period but were maintained in the placebo group. No effects of the antioxidant supplementation were observed in the erythrocyte antioxidant enzyme response to the exercise tests. The antioxidant supplementation induced a better adaptation of lymphocyte catalase after submaximal test. CONCLUSIONS: Lymphocytes showed higher sensibility to antioxidant supplementation, improving the response of antioxidant enzymes to training and to acute exercise. In erythrocytes the training adaptations were more important than the antioxidant supplementation effects. PMID- 16365697 TI - HIV knowledge and sexual risk behaviors of street children in Takoradi, Ghana. AB - A pilot survey was conducted among 100 street children aged 11-19 years and data collected regarding HIV knowledge, risk behaviors, home and sexual experiences, and factors that contributed to their relocation to the street environment. Data indicated that 80% of the street children had at least minimal knowledge of HIV and 54% perceived themselves to be at risk for contracting HIV. More than three fourths of the females, and over half of the boys reported ever having sex. There were significant differences between males and females for sexual activity, physical and sexual abuse, and prostitution, as girls were more likely to be sexually active, physically abused, and involved in prostitution. PMID- 16365698 TI - Synthesis and characterization of new fluorene-based singlet oxygen sensitizers. AB - The synthesis, photophysical characterization, and determination of singlet oxygen quantum yields (Phi(Delta)) for a class of fluorene derivatives with potential application in two-photon photodynamic therapy (PDT) is reported. It has been demonstrated that these compounds possess the ability to generate singlet oxygen (1O2) upon excitation. A photochemical method, using 1,3 diphenylisobenzofuran (DPBF) as 1O2 chemical quencher, was employed to determine the singlet oxygen quantum yields (Phi(Delta)) of the fluorene-based photosensitizers in ethanol. Phi(Delta) values ranged from 0.35 to 0.75. These derivatives may have potential application as two-photon photosensitizers when pumped via two-photon excitation in the near-IR spectral region. PMID- 16365699 TI - Can free energy calculations be fast and accurate at the same time? Binding of low-affinity, non-peptide inhibitors to the SH2 domain of the src protein. AB - The usefulness of free-energy calculations in non-academic environments, in general, and in the pharmaceutical industry, in particular, is a long-time debated issue, often considered from the angle of cost/performance criteria. In the context of the rational drug design of low-affinity, non-peptide inhibitors to the SH2 domain of the (pp60)src tyrosine kinase, the continuing difficulties encountered in an attempt to obtain accurate free-energy estimates are addressed. free-energy calculations can provide a convincing answer, assuming that two key requirements are fulfilled: (i) thorough sampling of the configurational space is necessary to minimize the statistical error, hence raising the question: to which extent can we sacrifice the computational effort, yet without jeopardizing the precision of the free-energy calculation? (ii) the sensitivity of binding free energies to the parameters utilized imposes an appropriate parametrization of the potential energy function, especially for non-peptide molecules that are usually poorly described by multipurpose macromolecular force fields. Employing the free energy perturbation method, accurate ranking, within +/-0.7 kcal/mol, is obtained in the case of four non-peptide mimes of a sequence recognized by the (pp60)src SH2 domain. PMID- 16365703 TI - Extraction of condensed tannins from cervid feed and feces and quantification using a radial diffusion assay. AB - A radial diffusion assay was employed to quantify condensed tannins (CT) in feed and feces of mule deer (Odocoileus hemionus) and white-tailed deer (O. virginianus). This biological assay measures the precipitation of bovine serum albumin (BSA), with the area of the precipitation ring being proportional to the amount of extracted CT applied to the well. CT extracted from the bark of white spruce (Picea glauca) was used as the standard. CT were extracted with 70% (v/v) aqueous acetone and precipitated with 50% (v/v) aqueous methanol or 70% (v/v) aqueous acetone. Functional range of CT weights for suitable ring measurement was 0.5-4.0 mg, and equilibrium was achieved within an incubation period of 24 hr. Methanol (50%) was a more effective precipitation solvent than acetone (70%) having 13 +/- 4% greater specific activity (P < 0.05) and superior capabilities for predicting CT content. Precipitation rings were evaluated on images magnified on a photocopier. Ring diameters measured on a 200% enlarged photocopy provided the most precise estimate of ring area (R2 = 0.98). This convenient method reduced analysis times and enhanced accuracy and precision of tannin quantification. Analytical consequences and future research requirements are considered. PMID- 16365704 TI - Rapid absorption of dietary 1,8-cineole results in critical blood concentration of cineole and immediate cessation of eating in the common brushtail possum (Trichosurus vulpecula). AB - The blood concentration of 1,8-cineole and its metabolites was measured in six male brushtail possums while they voluntarily fed on diets laced with varying concentrations of cineole for 3 d. On the third day, blood samples were collected during and after each bout of feeding for 3 hr. Blood cineole was measured by using headspace solid-phase microextraction (SPME), while cineole metabolites were measured by liquid-liquid extraction followed by gas chromatography-mass spectroscopy. Feeding patterns were measured by continual recording of residual food weight and time. Cineole absorption was rapid, resulting in a peak blood concentration at the end of each feeding bout. The blood concentration of cineole did not exceed a critical value (51.8 +/- 14.1 micromol/l) regardless of the concentration in the diet. Food and, therefore, cineole intake was regulated. The amount of food ingested in the first feeding bout decreased from 236 +/- 52 g on the control diet to 36 +/- 20 g on the 4% cineole diet. The amount of cineole ingested in the first bout (1.18 +/- 1.10 g) was the same regardless of the dietary concentration and was controlled by the size of the meal. Total food eaten during the 7-hr feeding session decreased by 64% from 368 +/- 94 g (control diet) to 131 +/- 52 g (4% diet). Total cineole intake increased from 2.47 +/- 0.60 g (1% diet) to 5.05 +/- 2.41 g (4% diet). Cineole metabolites accumulated throughout the sampling period and were generally still rising at the end of blood sampling period. Blood levels of metabolites were at least 10-fold higher than cineole levels. The immediate control of feeding seems to be regulated by blood levels of cineole, whereas metabolites are likely to be more important in regulating the chronic ingestion of cineole. PMID- 16365705 TI - Feeding responses of free-flying honeybees to secondary compounds mimicking floral nectars. AB - The role of secondary compounds (SC) in deterring herbivores and pathogens from vegetative parts of plants is well established, whereas their role in plant reproductive organs such as floral nectar is unclear. The present study aimed to reveal the response of free-flying honeybees to naturally occurring concentrations of four SC in floral nectar. We selected nicotine, anabasine, caffeine, and amygdalin, all of which are found in nectar of various plants. In repeated paired-choice experiments, we offered 20% sucrose solution as control along with test solutions of 20% sucrose with various concentrations of the above SC. Except for anabasine, naturally occurring concentrations of SC did not have a deterring effect. Furthermore, low concentrations of nicotine and caffeine elicited a significant feeding preference. SC can, therefore, be regarded as postingestive stimulants to pollinators, indicating that the psychoactive alkaloids in nectar may be a part of their mutualistic reward. Further studies are needed to test our hypothesis that psychoactive alkaloids in nectar impose dependence or addiction effects on pollinators. PMID- 16365706 TI - Nutrient composition of larval nectar secretions from three species of myrmecophilous butterflies. AB - A comparative chemical analysis of the larval nectar secretions and hemolymph from three unspecifically and facultatively ant-attended lycaenid species (Polyommatus coridon, P. icarus, and Zizeeria knysna) was performed by using high performance liquid chromatography techniques. Sucrose was the main sugar component in all three species. In half of the samples of P. coridon, it was accompanied by glucose, whereas other sugars occurred only rarely. In P. icarus and Z. knysna, melezitose was the second-most important component, followed by fructose and glucose. Total sugar contents were 43.6 +/- 14.8 g/l (mean +/- SD) for P. coridon, 74.2 g/l for P. icarus, and 68.3 +/- 22.6 g/l for Z. knysna. Up to 14 different identified amino acids were found in P. coridon nectar, with a total content of 9.7 +/- 3.4 g/l. Leucine was always the major component (contributing 50% of overall amino acid content). Other important amino acids were tyrosine, proline, arginine, and phenylalanine. P. icarus nectar contained up to six amino acids with a total content of 1.2 g/l, dominated by tyrosine and phenylalanine. Z. knysna nectar contained alanine and proline, with only 0.3 +/- 0.17 g/l total content. In the hemolymph of all species, up to 16 different amino acids occurred relatively regularly, with histidine dominating, followed by serine and proline. The amino acid pattern in hemolymph was considerably different from that of the nectar secretions. Larval diet weakly influenced P. coridon nectar sugars, and with a semisynthetic diet, a more homogeneous amino acid pattern was detected. Comparison with reports from other lycaenid species shows that secretions rich in amino acids are related to intimate, often obligate ant associations, whereas facultative, unspecific myrmecophiles rely on carbohydrates. PMID- 16365707 TI - The South African and Namibian populations of the resurrection plant Myrothamnus flabellifolius are genetically distinct and display variation in their galloylquinic acid composition. AB - The polyphenol contents and compositions in desiccated leaves of Myrothamnus flabellifolius plants collected in various locations in Namibia and South Africa were analyzed using UV spectroscopy and high-performance liquid chromatography mass spectrometry. A study of the genetic relatedness of these populations was also performed by determination of the DNA sequence of the intergenic spacer region between the psbA and the trnH genes in the chloroplast genome. Namibian M. flabellifolius plants contained significantly more polyphenols than South African plants. Namibian plants essentially contained a single polyphenol, 3,4,5-tri-O galloylquinic acid, whereas South African plants contained a variety of galloylquinic acids including 3,4,5-tri-O-galloylquinic acid together with higher molecular weight galloylquinic acids. Sequence analysis revealed a 1.4% divergence between Namibian and South African plants corresponding to the separation of these populations of approximately 4 x 10(6) years. The significance of the poly-phenol content and composition to the desiccation tolerance of the two populations is discussed. PMID- 16365708 TI - Phenological variation in chemical defense of the pipevine swallowtail, Battus philenor. AB - Larvae of the pipevine swallowtail, Battus philenor, feed on plants in the genus Aristolochia, which contains aristolochic acids, toxic alkaloids unique to the Aristolochiaceae. Pipevine swallowtails sequester these compounds and, as a consequence, are chemically defended against many natural enemies. In California, the primary aristolochic acid present in the butterfly is aristolochic acid I. Newly eclosed adult females possess greater amounts of these sequestered toxins compared to males. However, over the course of the flight season, the aristolochic acid content of females in the population declines, whereas male aristolochic acid content remains relatively constant. Transference of sequestered aristolochic acids to eggs by females might explain the decline of these sequestered chemical defenses observed over time. We found no evidence that males transfer aristolochic acids to females via the spermatophore. The possibility that females at the end of the flight season may be automimics of males is discussed. Temporal variation in the aristolochic acid defenses exhibited by this pipevine swallowtail population is both age- and sex-dependent. PMID- 16365709 TI - Calcium tartrate crystals in the midgut of the grape leafhopper. AB - Calcium tartrate crystals were observed in the midgut of grape leafhoppers. This unique compound was found for the first time in insects. The size of the crystals varied strongly between and within individuals with a mean length of 153 +/- 87 microm and a mean width of 71 +/- 46 microm. In addition, the number of crystals per individual showed a broad variation and ranged from 1 to 150 crystals/individual. The occurrence of calcium tartrate crystals as well as the number of crystals per individual followed the same seasonal pattern as seasonal vine leaf concentrations of tartaric acid found in a previous study, indicating that calcium tartrate is formed to neutralize the tartaric acid in the gut system. It further implies that the grape leafhopper, rather than being a pure phloem sucker, employs a mixed feeding strategy to satisfy its demands for calcium uptake. PMID- 16365710 TI - Chemically mediated host-plant selection by the milfoil weevil: a freshwater insect-plant interaction. AB - The milfoil weevil Euhrychiopsis lecontei is a specialist aquatic herbivore that feeds, oviposits, and mates on the invasive freshwater macrophyte Myriophyllum spicatum. We characterized the weevil's preference for M. spicatum, and through bioassay-driven fractionation, isolated and identified two chemicals released by M. spicatum that attract E. lecontei. Mass spectrometry and nuclear magnetic resonance spectroscopy were used to identify the attractive compounds as glycerol and uracil. Dose-response curves for glycerol and uracil indicated that weevil preference increased as sample concentration increased. Weevils were attracted to a crude sample of M. spicatum-released chemicals from 0.17 to 17 mg/l, to glycerol from 18 to 1800 microM (0.0017-0.17 mg/l), and to uracil from 0.015 to 15 microM (0.00014-1.4 mg/l). Although glycerol and uracil are ubiquitous, weevils are likely responding to high concentrations that are released as a result of the rapid growth of M. spicatum. Uracil concentration was greater in the exudates of M. spicatum than other Myriophyllum spp. E. lecontei was attracted to glycerol at a concentration similar to that at which terrestrial insects are attracted to sugar alcohols. This is the first example of a freshwater specialist insect being attracted to chemicals released by its host plant. Analysis of the water milfoil-weevil interaction provides further understanding as to how insects locate their host plants in aquatic systems. PMID- 16365711 TI - Unraveling the nature of individual recognition by odor in hermit crabs. AB - Individual recognition is a key element in the social life of many invertebrates. However, most studies conducted so far document that several species are capable of a "binary" discrimination among conspecifics, but not of a "true individual recognition." Our objective was to learn more about the mechanisms that underlie individual recognition by odor in hermit crabs by individuating some of its properties. Using Pagurus longicarpus Say 1817 as a model species, we conducted four series of experiments in which the response of every test crab (the "receiver") to the different odor treatments (emitted by a "sender") was evaluated from its investigative behavior toward an empty, high-quality shell. After having excluded the possibility that crabs chemically recognize familiar/unfamiliar shells and/or shells of high/low quality, we explored whether the receivers discriminate odors from two familiar senders and whether this discrimination also occurs with unfamiliar crabs. We also asked whether crabs form an association between the odor of a familiar sender and some of its relevant attributes, i.e., rank, size, and shell quality. Finally, the shells inhabited by familiar individuals were manipulated to modify the association between odor and shell quality. Results showed that: (1) there is no odor specific of a rank; (2) individual crabs discriminate their own odor from the odor of other individuals; (3) they can chemically discriminate between larger crabs inhabiting higher-quality shells and smaller crabs inhabiting lower-quality shells, provided that these crabs are familiar to them; (4) they associate the odor of an individual crab with the quality of the shell it inhabits; and (5) this association quickly changes when social partners switch to shells of different quality. These results indicate that the nature of chemical recognition in P. longicarpus is more refined than a simple binary system. The receiver appears able to associate a type of information from the sender with memories of past experiences, therefore suggesting the hermit crab's potential for relatively high-order knowledge about conspecifics. PMID- 16365712 TI - Pheromone components from body scales of female Anarsia lineatella induce contacts by conspecific males. AB - Pheromonal communication of adult peach twig borers, Anarsia lineatella Zeller (Lepidoptera: Gelechiidae), was reinvestigated based on recent findings that virgin female-baited traps were more attractive to mate-seeking males than a two component synthetic sex pheromone consisting of (E)-5-decen-1-yl acetate (1000 microg) and (E)-5-decen-1-ol (100 microg), suggesting that females use additional pheromone components. Hypothesizing that these additional components may be released from body parts other than abdominal sex pheromone glands, we extracted female body scales and analyzed aliquots by coupled gas chromatographic electroantennographic detection (GC-EAD) and GC-mass spectrometry. Eight straight chain and four methylated aliphatic hydrocarbons, as well as two acetates, all elicited responses from excised male antennae. In laboratory experiments with synthetic candidate pheromone components, a combination of octadecyl acetate, (R) 11-methyltricosane, and (S)-11-methyltricosane in the presence of gland-derived sex pheromone components were shown to elicit contact of female decoys by males. However, body pheromone components did not enhance attractiveness of sex pheromone components in field trapping experiments, suggesting that they are effective only at close range and that other stimuli are responsible for superior attractiveness of female-baited traps. PMID- 16365713 TI - Intersexual differences in chemical composition of precloacal gland secretions of the amphisbaenian Blanus cinereus. AB - Pheromonal communication seems to be especially important in amphisbaenians, a group of reptiles morphologically and functionally adapted to fossorial life. Both male and female amphisbaenians (Blanus cinereus) produce copious amounts of a secretion from the precloacal glands. Analysis by gas chromatography-mass spectrometry revealed that these secretions contain 29 major lipophilic compounds, including several steroids (mainly cholesterol and cholesteryl methyl ether), n-C9 to n-C18 carboxylic acids (more diverse in males), and methyl and long-chain (waxy)-type esters of carboxylic acids, along with squalene (mainly in males) and tocopherol (only in females). There were clear intersexual differences in the presence/absence of some compounds, such as some fatty acids, steroids, and tocopherol, and in the relative proportions of some shared compounds, such as squalene. These differences may explain how the sexes of this amphisbaenian discriminate one another based on scent alone. The abundance of steroids and waxy esters in secretions of both male and female amphisbaenians may be useful to scent mark underground tunnels to facilitate orientation by these organisms. PMID- 16365714 TI - New pheromone components of the grapevine moth Lobesia botrana. AB - Analysis of extracts of sex pheromone glands of grapevine moth females Lobesia botrana showed three previously unidentified compounds, (E)-7-dodecenyl acetate and the (E,E)- and (Z,E)-isomers of 7,9,11-dodecatrienyl acetate. This is the first account of a triply unsaturated pheromone component in a tortricid moth. The monoenic acetate (E)-7-dodecenyl acetate and the trienic acetate (7Z,9E,11) dodecatrienyl acetate significantly enhanced responses of males to the main pheromone compound, (7E,9Z)-7,9-dodecadienyl acetate, in the wind tunnel. The identification of sex pheromone synergists in L. botrana may be of practical importance for the development of integrated pest management systems. PMID- 16365715 TI - (S,S)-2,12-, (S,S)-2,13-, and (S,S)-2,14-diacetoxyheptadecanes: sex pheromone components of red cedar cone midge, Mayetiola thujae. AB - We identified, synthesized, and field-tested the sex pheromone of female red cedar cone midge Mayetiola thujae (Hedlin) (Diptera: Cecidomyiidae), a pest insect in red cedar Thuja plicata seed orchards. Coupled gas chromatographic (GC) electroantennographic detection analyses of pheromone extract revealed three components (A, B, C) that elicited responses from antennae of males, all of which occurred below the detection threshold of the mass spectrometer and thus had to be identified without spectroscopic data. Taking into account (1) their retention indices (RI) on three GC columns (DB-5, DB-23, and DB-210), (2) intercolumn RI differentials, and (3) the molecular structures of known cecidomyiid pheromones, we synthesized seven candidate pheromone components: 2,10-, 2,11-, 2,12-, 2,13-, 2,14-, 2,15- and 2,16-diacetoxyheptadecanes. Of these, 2,12-, 2,13-, and 2,14 diacetoxyheptadecane had RIs on all columns consistent with those of A, B, and C and elicited strong antennal responses when tested at picogram levels. In field experiments with the twelve stereoselectively synthesized stereoisomers, only the SS-stereoisomers of 2,12-, 2,13-, and 2,14-diacetoxyheptadecane attracted male M. thujae. The three-component SS-stereoisomer blend was more attractive than the 12 component blend of all stereoisomers, suggesting that one or several nonnatural stereoisomers are inhibitory. One-, two-, and three-component lures of the SS stereoisomers were equally effective in attracting male M. thujae, indicating redundancy in the pheromone. Identification of the M. thujae sex pheromone will allow development of pheromone-based monitoring, and possibly control, of M. thujae populations in T. plicata seed orchards. PMID- 16365716 TI - Electrophysiological and behavioral responses to chocolate volatiles in both sexes of the pyralid moths Ephestia cautella and Plodia interpunctella. AB - Volatiles from chocolate mediate upwind flight behavior in Ephestia cautella and Plodia interpunctella. We used gas chromatography with electroantennographic detection and found 12 active compounds derived from three different chocolate types, i.e., plain, nut-containing, and rum-flavored. Eight of the compounds were identified with mass spectrometry, and the activity of three compounds, ethyl vanillin, nonanal, and phenylacetaldehyde (PAA), was subsequently confirmed in both electrophysiological and behavioral assays. In the electroantennogram experiment, PAA and nonanal were consistently eliciting responses in both species and sexes. Ethyl vanillin was active in males of both species, and also in P. interpunctella females. E. cautella females showed no antennal activity in response to ethyl vanillin. All three volatiles were attractive to E. cautella males and P. interpunctella females in a flight tunnel. E. cautella females were significantly attracted only to ethyl vanillin. P. interpunctella males were attracted to PAA. Ethyl vanillin is a novel insect attractant, whereas both nonanal and phenylacetaldehyde mediate behavior in many insect species. A final experiment revealed that a blend of the three volatiles was required to induce landing in the flight tunnel bioassay, and that the landing rate was dependent on dose. The three-component blend attracted both sexes of P. interpunctella and females of E. cautella, whereas E. cautella males were not attracted. PMID- 16365717 TI - Phytotoxicity of Phytolacca americana leaf extracts on the growth, and physiological response of Cassia mimosoides. AB - We examined the allelochemical effects of control soil, native soil (treated soil), and leaf extracts of Phytolacca americana (pokeweed) on the germination rate and seedling growth of Cassia mimosoides var. nomame. We also studied the resulting changes in root-tip ultrastructure and peroxidase isozyme biochemistry. P. americana leaf extract inhibited seed germination, seedling growth, and biomass when compared to control and treated soil. Root and shoot growth in treated soil was stimulated relative to control soil, but root growth was inhibited by 50% in the leaf extract treatment. Biomass of C. mimosoides seedlings grown on leaf extract was reduced sevenfold when compared to the control seedlings. The amounts of total phenolic compounds in the leaf extract, treated soil, and control soil were 0.77, 0.14, and 0.03 mg l(-1), respectively. The root tips of C. mimsoides treated with leaf extracts of P. americana showed amyloplasts and large central vacuoles with electron-dense deposits inside them when compared to control root tips. The activity of guaiacol peroxidase (GuPOX) in whole plant, roots, and shoots of C. mimosoides increased as leaf extract increased; maximum activity was observed in extract concentrations of 75% and higher. Root GuPOX activity was three times higher than in shoots. Therefore, we conclude that inhibition of C. mimosoides growth is related to the phenolic compounds in the P. americana leaf extract and the ultrastructure changes in root tip cells and increased GuPOX activity is a response to these allelochemicals. PMID- 16365718 TI - Chemistry of the lichen Hypogymnia physodes transplanted to an industrial region. AB - Lichens produce a great number of secondary metabolites that participate in ecological interactions and respond to environmental changes. We examined the influence of heavy metal accumulations on lichen secondary metabolism. Thalli of Hypogymnia physodes were transplanted for 6 months to the Cracow-Silesia industrial region. Based on heavy metal accumulations in lichen, two of the investigated sites were classified as highly polluted. The highest concentrations of Cd, Pb, and Zn were found in lichens transplanted in the vicinity of a Zn-Pb smelter. Significant accumulations of Cr and Ni were detected in Hypogymnia transplanted near a chemical industry. Physodic, physodalic, hydroxyphysodic acids, and atranorin were identified and analyzed in extracts obtained from specimen samples. The most detrimental changes were observed in lichen transplanted into the vicinity of a chemical industry producing chromium, phosphor, and sulfur compounds that contained 340-fold higher Cr levels than control thalli. Decreases in the levels of physodic acid, hydroxyphysodic acid, and atranorin were detected, and one additional polar compound (probably product of degradation of lichen acids) appeared in the extract. The content of physodalic acid increased in every thalli sample transplanted, suggesting a possible role of this compound in defense against stress caused by accumulated pollutants. The levels of physodic acid decreased in thalli from both of the most polluted sites compared to those of the controls--but were not changed in thalli transplanted to less polluted sites. Our results illustrate that lichen compounds are sensitive to heavy metal accumulation and could be used as biomarkers in environmental studies. PMID- 16365721 TI - Healing of intrabony defects following treatment with an oily calcium hydroxide suspension (Osteoinductal). A controlled clinical study. AB - The purpose of the present clinical study was to evaluate the healing of deep intrabony defects following the application of an oily calcium hydroxide suspension (OCHS). Thirty patients suffering from chronic periodontitis, each of whom displayed one intrabony defect, were randomly treated with access flap surgery (AFS) and the application of OCHS (test) or with AFS alone (control). The following clinical parameters were recorded at baseline and at 6 months after therapy: plaque index, gingival index, bleeding on probing, probing depth (PD), gingival recession, and clinical attachment level (CAL). No differences in any of the investigated parameters were observed at baseline between the two groups. At 6 months after therapy, the test group showed a reduction in mean PD from 7.7+/ 1.5 to 2.9+/-0.9 mm (P<0.001) and a change in mean CAL from 9.6+/-2.1 to 5.5+/ 2.5 mm (P<0.001). In the control group, the mean PD was reduced from 6.9+/-0.9 to 3.7+/-0.9 mm (P<0.001) and the mean CAL changed from 8.5+/-2.5 to 6.4+/-2.7 mm (P<0.001). OCHS resulted in statistically significant higher PD reductions (P<0.01) and CAL gains (P<0.05) than AFS alone. Within the limits of the present study, it can be concluded that: (1) at 6 months after surgery both therapies resulted in statistically significant PD reductions and CAL gains and (2) treatment with OCHS resulted in statistically significant higher CAL gains than treatment with AFS alone. PMID- 16365722 TI - Successful treatment of fungus balls due to fluconazole-resistant Candida sake obstructing ureter stents in a renal transplant patient. AB - Reported here is the case of a 72-year-old renal transplant recipient with stenosis of the neo-ureter requiring stents, who was admitted to hospital with pyonephrosis caused by fungus balls. Fluconazole-resistant Candida sake was grown. Treatment with external drainage of the renal pelvis and intravenous and local administration of caspofungin resulted in relief of the obstruction. Eradication of the infection was achieved by surgical removal of the ureter with all stents and construction of a cysto-pyelostomy. PMID- 16365723 TI - Caspofungin treatment of Pneumocystis pneumonia during conditioning for bone marrow transplantation. PMID- 16365724 TI - Long-term mortality in a nationwide cohort of childhood-onset type 1 diabetic patients in Norway. AB - AIMS/HYPOTHESIS: We examined long-term total and cause-specific mortality in a nationwide, population-based Norwegian cohort of patients with childhood-onset type 1 diabetes. MATERIALS AND METHODS: All Norwegian type 1 diabetic patients who were diagnosed between 1973 and 1982 and were under 15 years of age at diagnosis were included (n=1,906). Mortality was recorded from diabetes onset until 31 December 2002 and represented 46,147 person-years. The greatest age attained among deceased subjects was 40 years and the maximum diabetes duration was 30 years. Cause of death was ascertained by reviews of death certificates, autopsy protocols and medical records. The standardised mortality ratio (SMR) was based on national background statistics. RESULTS: During follow-up 103 individuals died. The mortality rate was 2.2/1000 person-years. The overall SMR was 4.0 (95% CI 3.2-4.8) and was similar for males and females. For ischaemic heart disease the SMR was 20.2 (7.3-39.8) for men and 20.6 (1.8-54.1) for women. Acute metabolic complications of diabetes were the most common cause of death under 30 years of age (32%). Cardiovascular disease was responsible for the largest proportion of deaths from the age of 30 years onwards (30%). Violent death accounted for 28% of the deaths in the total cohort (35% among men and 11% among women). CONCLUSIONS/INTERPRETATION: Childhood-onset type 1 diabetes still carries an increased mortality risk when compared with the general population, particularly for cardiovascular disease. To reduce these deaths, attention should be directed to the prevention of acute metabolic complications, the identification of psychiatric vulnerability and the early detection and treatment of cardiovascular disease and associated risk factors. PMID- 16365725 TI - The effect of intrauterine environment and low glomerular number on the histological changes in diabetic glomerulosclerosis. AB - AIMS/HYPOTHESIS: We tested the hypothesis that diabetic glomerulosclerosis would develop more rapidly in animals with fewer glomeruli. METHODS: We studied the female offspring of Wistar rats that had been fed a low-protein diet (LPD) containing 6% protein or a normal-protein diet (NPD) containing 18% protein during pregnancy. Streptozotocin diabetes was induced at 12 weeks and animals were killed at 40 weeks. RESULTS: Non-diabetic LPD offspring were of lower birthweight than the NPD offspring (5.19+/-0.64 vs 6.45+/-0.67 g, p<0.001) and had fewer glomeruli (27,402+/-3,137 vs 34,203+/-6,471, p<0.05). Glomerular volume correlated inversely with glomerular number (r=-0.64, p=0.035), but total glomerular filtration surface area was reduced in the LPD animals (4,770+/-541 vs 5,779+/-1,302 mm(2), p=0.05). Other renal structural and functional parameters were similar. In LPD and NPD diabetic animals, glomerular volume and basement membrane width were significantly increased compared to their respective controls. Podocyte density was lowest in the LPD diabetic animals (not significant), and the area covered by each podocyte was greater in the LPD diabetic group (2.40+/-0.693 x10(-3) mm(2)) than in the LPD control group (1.68+/ 0.374 x10(-3) mm(2), p<0.001) and in the NPD diabetic animals (1.71+/-0.291 x 10( 3) mm(2), p<0.05). There was no difference in any other structural or functional parameter between the LPD and NPD diabetic animals. CONCLUSIONS/INTERPRETATION: A decrease in glomerular number was not deleterious to renal structure and function over 40 weeks in this animal model. Further work in models with progressive renal impairment and hypertension is necessary to clarify the impact of glomerular number on the development of renal disease. PMID- 16365726 TI - Proteinuria and metabolic syndrome as predictors of cardiovascular death in non diabetic and type 2 diabetic men and women. AB - AIMS/HYPOTHESIS: Proteinuria predicts cardiovascular disease (CVD), but it is unclear whether this is explained by the association of the metabolic syndrome with proteinuria. Therefore, we investigated proteinuria and the metabolic syndrome as independent predictors of CVD death in men and women. METHODS: The cohort comprised 574 non-diabetic men, 707 non-diabetic women, 371 diabetic men and 349 diabetic women, all free of CVD at baseline. Modified World Health Organization criteria were used to define the metabolic syndrome, and a urinary protein concentration of >or=0.1 g/l (or >or=0.2 g/l) to define proteinuria. The endpoint was CVD mortality during the 18-year follow-up. RESULTS: Among non diabetic men, CVD mortality per 1,000 person-years was as follows: no metabolic syndrome, no urinary protein group: 5.3; no metabolic syndrome, positive for urinary protein: 8.9; positive for metabolic syndrome, no urinary protein: 13.3; and positive for metabolic syndrome and urinary protein: 14.9. For non-diabetic women the corresponding values were: 0.9, 2.3, 4.9 and 7.9, respectively. Among diabetic men, CVD mortality per 1,000 person-years was 15.2, 32.5, 23.6 and 42.0 for the respective groups. Among diabetic women it was 25.3, 38.0, 26.3 and 40.3 (urinary protein in all cases defined as >or=0.1 g/l). In multivariate Cox models including both urinary protein and metabolic syndrome, the hazard ratios (HRs, 95% CI) of proteinuria for CVD mortality were 1.5 (0.9-2.4) in non-diabetic men, 1.8 (0.8-4.2) in non-diabetic women, 1.6 (1.0-2.6) in diabetic men and 1.6 (1.1 2.3) in diabetic women. Urinary protein as a continuous variable was associated with CVD mortality in all groups. The corresponding HRs for metabolic syndrome were: 1.6 (0.9-2.7), 4.0 (1.7-9.7), 1.5 (1.1-2.0) and 1.1 (0.8-1.5). CONCLUSIONS/INTERPRETATION: Proteinuria predicted CVD mortality independently of the presence of metabolic syndrome in non-diabetic and diabetic subjects. Metabolic syndrome predicted CVD mortality in non-diabetic women and in diabetic men, independently of the presence of proteinuria. PMID- 16365727 TI - Oncocytic modifications in rectal adenocarcinomas after radio and chemotherapy. AB - The purpose of the study is to highlight oncocytic modifications in rectal adenocarcinomas and evaluate a possible correlation with preoperative radiochemotherapy (RCT). Twenty-eight cases of advanced rectal carcinoma, treated preoperatively by 5-fluorouracil (200-225 mg/m(2)) and 44-46 Gy in 22-23 fractions, were studied. All patients underwent biopsy before RCT. Surgery was performed within 6 weeks after RCT. In all cases oncocytic modifications were searched for on hematoxylin and eosin (H&E) and at immunohistochemistry using an antimitochondrial antibody. In addition, in two cases, both pre- and post-RCT tissues were examined at electron microscopy. All tumors were adenocarcinomas. In pre-RCT biopsies, oncocytic changes were difficult to find on H&E, while the antimitochondrial antibody strongly stained numerous neoplastic cells (mean 48.4%). In post-RCT surgical specimens, oncocytic changes were detected in 24 out of 28 cases on H&E and the antimitochondrial antibody stained most of the residual neoplastic cells (mean 76.7%). Ultrastructural examination revealed large and bizarre mitochondria inside tumor cells both in pre- and post-RCT tissues. In conclusion, the present data suggest that rectal adenocarcinomas are "mitochondrion-rich" tumors. After preoperative RCT, residual neoplastic cells acquire a definite oncocytic phenotype. PMID- 16365728 TI - Re: Detection of JC virus sequences in colorectal cancers in Japan. PMID- 16365729 TI - Rhabdomyosarcoma: molecular diagnostics of patients classified by morphology and immunohistochemistry with emphasis on bone marrow and purged peripheral blood progenitor cells involvement. AB - Two histologically distinct subtypes of rhabdomyosarcomas (RMS), embryonal and alveolar, are different in many aspects, such as age distribution, primary site, and clinical outcome. We analyzed a group of 30 patients with RMS. The aim was to broaden the spectrum of diagnostic tools in evaluating the primary tumors, their recurrences and/or metastases, and to extend the diagnostic boundary to bone marrow and purged peripheral progenitor blood cell samples. We have performed the RT-PCR assay to analyze RMS for the presence of expression of MyoD1 gene and for the presence of chimeric transcripts PAX3/FKHR or PAX7/FKHR. MyoD1 gene expression was found in all 30 patients in samples from primary tumors. The chimeric transcripts PAX/FKHR were identified in 13 of 15 patients with alveolar RMS. Furthermore, the fusion transcript PAX7/FKHR was identified in 2 of 15 patients with RMS classified as embryonal by histology. Bone marrow samples (12) and peripheral blood progenitor cell specimens (13) in ten patients were examined by RT-PCR. We were able to identify 7 patients with bone marrow involvement and/or with contamination of peripheral blood progenitor cells by the tumor cells. We demonstrate that employing molecular diagnostics has an impact on staging, therapy monitoring and recognition of malignant cells at the tumor resection margins. PMID- 16365730 TI - False-positive FDG PET uptake--the role of PET/CT. AB - Positron emission tomography (PET) is a powerful molecular imaging technique for the human body-imaging applications currently available. As altered glucose metabolism is characteristic for many malignancies, FDG-PET is mostly used in oncology for staging and therapy control. Although PET is a sensitive tool for detecting malignancy, FDG uptake is not tumor specific. It can also be seen in healthy tissue or in benign disease as inflammation or posttraumatic repair and could be mistaken for cancer. The experienced nuclear medicine physician mostly manages to differentiate malignant from non-malignant FDG uptake, but some findings may remain ambiguous. In these cases, the difficulties in differentiating physiologic variants or benign causes of FDG uptake from tumor tissue can often be overcome by combined PET and CT (PET/CT) as anatomic information is added to the metabolic data. Thus, PET/CT improves the diagnostic accuracy compared to PET alone and helps to avoid unnecessary surgery/therapy. However, PET/CT involves other sources of artifacts that may occur when using CT for attenuation correction of PET or by patient motion caused by respiration or bowel movements. PMID- 16365731 TI - FLAIR imaging for multiple sclerosis: a comparative MR study at 1.5 and 3.0 Tesla. AB - The purpose of this study was (1) to identify the optimal TE for FLAIR-imaging at 3.0 T assessing three different echo times qualitatively and quantitatively and (2) to evaluate the diagnostic efficacy of high-field 3.0-T FLAIR imaging in comparison to conventional 1.5-T MRI in patients with multiple sclerosis (MS). Twenty-two patients with clinically definite MS underwent axial FLAIR imaging at 1.5 and 3.0 T. In 15 of these patients further FLAIR images with a TE of 100, 120 and 140 ms were acquired at 3.0 T. Imaging protocols were modified for 3.0 T using the increased SNR to acquire more and thinner slices while maintaining a comparable scan time. FLAIR images of either different TEs or different field strengths were ranked for each patient qualitatively by two observers. Signal intensity measurements were obtained in the gray and white matter, CSF and representative white matter lesions (WML). At 3.0 T, a TE of 100 and 120 ms proved superior in all qualitative categories when compared to 140 ms. In the quantitative assessment CNR of WML was highest for 120 ms (CNR: 19.8), intermediate for 100 ms (17.2) and lowest for 140 ms (15.3) (P<0.003). For lesion conspicuity and overall image quality, 3.0 T was judged superior to 1.5 T, whereas no difference was found for gray-white differentiation and image noise. With regard to artifacts, 3.0 T was inferior to 1.5 T. The CNR for WML was slightly lower at 3.0 T, but the difference was not significant (22.6 vs. 28.0, P=ns). However, significantly more WML were detected at 3.0 T than at 1.5 T (483 vs. 341, P<0.0001). The optimal echo time for FLAIR imaging at 3.0 T is 120 ms due to the significantly higher CNR of WML. By trading the higher SNR at 3.0 T for better spatial resolution, nearly the same CNR level could be maintained, increasing lesion detectability at 3.0 T compared to 1.5 T. Thus, high-field MRI may further strengthen the role of MRI as the most sensitive paraclinical test for the early diagnosis of MS. PMID- 16365732 TI - [The dry eye. Current concepts on classification, diagnostics, and pathogenesis]. AB - The etiology of dysfunctional tear syndrome ("dry eye") is multifactorial and complex. Recent evidence suggests an important role of androgens in regulating tear film secretion onto the ocular surface. In addition, inflammatory processes of the ocular surface seem to be the common final pathway of all chronic forms of dry eye. Novel concepts of pathogenesis and state-of-the-art diagnostic tools are discussed. In vivo confocal microscopy allows quantification of ocular surface inflammatory cells. This is of increasing importance for evaluation of anti inflammatory treatments in dry eye patients. PMID- 16365733 TI - Analysis of avian bone response to mechanical loading. Part two: Development of a computational connected cellular network to study bone intercellular communication. AB - Mechanical loading-induced signals are hypothesized to be transmitted and integrated by connected bone cells before reaching the bone surfaces where adaptation occurs. A computational connected cellular network (CCCN) model is developed to explore how bone cells perceive and transmit the signals through intercellular communication. This is part two of a two-part study in which a CCCN is developed to study the intercellular communication within a grid of bone cells. The excitation signal was computed as the loading-induced bone fluid shear stress in part one. Experimentally determined bone adaptation responses (Gross et al. in J Bone Miner Res 12:982-988, 1997 and Judex et al. in J Bone Miner Res 12:1737-1745, 1997) are correlated with the fluid shear stress by the CCCN, which adjusts cell sensitivities (loading and signal thresholds) and connection weights. Intercellular communication patterns extracted by the CCCN indicate the cell population responsible for perceiving the loading-induced signal, and loading threshold is shown to play an important role in regulating the bone response. PMID- 16365734 TI - Multiscale analysis of the coupled effects governing the movement of interstitial fluid in cortical bone. AB - A multiscale approach (periodic homogenization) is carried out to model osteon's behaviour, and especially the coupled phenomena that govern its interstitial fluid movement. Actions of electro-osmotic and osmotic motions in addition to the classical Poiseuille flow are studied at the mesoscale of the canaliculus and within the micropores of the collagen-apatite matrix. Use of this fully coupled modelling leads to a comparison of these different effects. Limitation of a classical Darcian description of the fluid flow at the two scales is so studied. For each of these studies a special attention is given to the pore's geometry influence and to their electrical and hydraulic properties. PMID- 16365735 TI - Emergency treatment of eye burns: which rinsing solution should we choose? AB - BACKGROUND: In the treatment of eye burns few data on the comparative application of rinsing solutions exist. We present experiments in vitro and ex vivo on the pH changes that can be achieved in alkali eye burns with currently distributed and propagated rinsing fluids like water, saline solution, Cederroth Eye Wash Solution (including borate buffer), Diphoterine, Ringers lactate solution and phosphate buffer. METHODS: Titration curves in beakers are compared with ex vivo experiments on isolated rabbit eyes. We exposed eyes to burns from filter paper soaked in 2 mol NaOH, continuously measuring the anterior chamber pH by means of a micro pH electrode placed near the endothelium. In each experiment--repeated five times--the corneal burn of 20 s in 2 mol NaOH was followed by a period of 15 min of rinsing under a defined flow of 66 ml/min. RESULTS: We found highly significant differences in intracameral pH related to different types of rinsing solutions. The return of the intracameral pH to normal was not achieved by any of the rinsing fluids, but the best results were noted for of Cederroth Eye Wash Solution (Cederroth Industrial Products, Upplands Vaasby, Sweden) and the Diphoterine- and Previn solutions (Prevor, Cologne, Germany). Water played an intermediate role whereas saline and phosphate buffer were not efficient at lowering intracameral pH after alkali burns. CONCLUSION: In alkali burns we recommend efficient buffering solutions. The tested isotonic phosphate buffer (PBS) was not effective at buffering the intraocular pH. Water was found to be much less efficient than Previn, Diphoterine or Cederroth Eye Wash solution in balancing intraocular pH. PMID- 16365736 TI - Vitreous prolapse and IOL dislocation during intravitreal injection of triamcinolone acetonide. AB - BACKGROUND: To report on procedure-related anterior segment complications during intravitreal injections. METHODS: In a prospective interventional case series, 614 eyes received a total of 723 intravitreal injections of about 20 mg triamcinolone acetonide (in 0.2 ml) after paracentesis and aqueous humor drainage for various indications. RESULTS: In three eyes (0.49% of all eyes) a vitreous prolapse occurred during the injection. In one eye, the vitreous prolapse was combined with dislocation of the intraocular lens (IOL). All three eyes were pseudophakic, showing an posterior capsule defect, and the IOL located in the ciliary sulcus. They were treated by translimbal vitrectomy, and one eye with reposition of the IOL. No other procedure-related postoperative complications were observed during injection or follow-up (7.8+/-7.1 months). CONCLUSIONS: Intravitreal injections may cause a vitreous prolapse into the anterior chamber with or without IOL decentration or dislocation in predisposed eyes. Ophthalmologists should be aware of this possible complication and inform patients at risk. PMID- 16365737 TI - Safety profile of the intravitreal streptokinase-plasmin complex as an adjunct to vitrectomy in the rabbit. AB - BACKGROUND: The generation of an atraumatic posterior vitreous detachment (PVD), a common goal in vitreoretinal surgery, is a challenge, particularly in children and young trauma patients. Plasmin has been proposed as an adjunct to vitrectomy to enzymatically generate a PVD. Low doses of streptokinase-activated plasmin were tested in human pilot studies. This dose-escalation study assesses the safety range of intravitreal human streptokinase-plasmin in rabbits. METHODS: Plasminogen was isolated from human plasma by affinity chromatography, followed by activation with streptokinase (1:1), to generate the streptokinase-plasmin complex. Enzyme doses from 0.1-7 activity units (AU, in 0.1 ml) were injected into the mid-vitreous of 35 eyes; six control eyes were injected with balanced salt solution (BSS, 0.1 ml). Thirty minutes after injection, a two-port vitrectomy was performed. Fundus and slit lamp examinations were performed on days 1 and 7. On days 2 and 7, bright flash electroretinography was performed and compared with preoperative recordings. Some animals receiving higher doses of streptokinase-plasmin (1-7 AU) were followed clinically and with electroretinography for up to 9 months. RESULTS: A mild-to-moderate inflammatory response was seen in both control and plasmin-treated eyes on day 1, but had disappeared completely by day 7 in most eyes. In the 7 AU group, inflammation was stronger and more protracted. Two of three eyes from this group developed wrinkling of the medullary rays; one of them showed discoloration and traction at the medullary rays in the late follow-up. Electroretinograms (ERGs) of vitrectomized control eyes showed the following changes from preoperative values: 48 h, a-wave -11.10% [no significant (n.s.)], b-wave -14.62% (P=0.046); 7 days, a wave +9.18% (n.s.), b wave +11.69% (n.s.). For the enzyme-treated eyes: 48 h: a wave -20.43% (P<0.001), b-wave -9.57% (p<0.001); 7 days: a wave -14.21% (P<0.001), b-wave +2.48% (P<0.001). There was no evidence of dose-dependent ERG changes in enzyme-treated eyes at doses up to 5 AU. Groups of up to 3 AU were investigated by light and transmission electron microscopy, without evidence of toxicity. CONCLUSION: Streptokinase-plasmin doses up to 3 AU were found to be safe when injected into rabbit eyes followed by vitrectomy. PMID- 16365738 TI - Genuine modern analogues of Precambrian stromatolites from caldera lakes of Niuafo'ou Island, Tonga. AB - Calcareous or dolomitic, often secondarily silicified, laminated microbial structures known as stromatolites are important keys to reconstruct the chemical and biotic evolution of the early ocean. Most authors assume that cyanobacteria associated microbialitic structures described from Shark Bay, Western Australia, and Exuma Sound, Bahamas, represent modern marine analogues for Precambrian stromatolites. Although they resemble the Precambrian forms macroscopically, their microstructure and mineralogical composition differ from those characterizing their purported ancient counterparts. Most Precambrian stromatolites are composed of presumably in situ precipitated carbonates, while their assumed modern marine analogues are predominantly products of accretion of grains trapped and bound by microbial, predominantly cyanobacterial, benthic mats and biofilms and only occasionally by their physicochemical activity. It has therefore been suggested that the carbonate chemistry of early Precambrian seawater differed significantly from modern seawater, and that some present-day quasi-marine or non-marine environments supporting growth of calcareous microbialites reflect the hydrochemical conditions controlling the calcification potential of Precambrian microbes better than modern seawater. Here we report the discovery of a non-marine environment sustaining growth of calcareous cyanobacterial microbialites showing macroscopic and microscopic features resembling closely those described from many Precambrian stromatolites. PMID- 16365739 TI - A novel role for proline in plant floral nectars. AB - Plants offer metabolically rich floral nectar to attract visiting pollinators. The composition of nectar includes not only sugars, but also amino acids. We have examined the amino acid content of the nectar of ornamental tobacco and found that it is extremely rich (2 mM) in proline. Because insect pollinators preferentially utilize proline during the initial phases of insect flight and can reportedly taste proline, we determined whether honeybees showed a preference for synthetic nectars rich in proline. We therefore established an insect preference test and found that honeybees indeed prefer nectars rich in the amino acid proline. To determine whether this was a general phenomenon, we also examined the nectars of two insect-pollinated wild perennial species of soybean. These species also showed high levels of proline in their nectars demonstrating that plants often produce proline-rich floral nectar. Because insects such as honeybees prefer proline-rich nectars, we hypothesize that some plants offer proline-rich nectars as a mechanism to attract visiting pollinators. PMID- 16365740 TI - Predicting memory performance in normal ageing using different measures of hippocampal size. AB - A number of different methods have been employed to correct hippocampal volumes for individual variation in head size. Researchers have previously used qualitative visual inspection to gauge hippocampal atrophy. The purpose of this study was to determine the best measure(s) of hippocampal size for predicting memory functioning in 102 community-dwelling individuals over 80 years of age. Hippocampal size was estimated using magnetic resonance imaging (MRI) volumetry and qualitative visual assessment. Right and left hippocampal volumes were adjusted by three different estimates of head size: total intracranial volume (TICV), whole-brain volume including ventricles (WB+V) and a more refined measure of whole-brain volume with ventricles extracted (WB). We compared the relative efficacy of these three volumetric adjustment methods and visual ratings of hippocampal size in predicting memory performance using linear regression. All four measures of hippocampal size were significant predictors of memory performance. TICV-adjusted volumes performed most poorly in accounting for variance in memory scores. Hippocampal volumes adjusted by either measure of whole-brain volume performed equally well, although qualitative visual ratings of the hippocampus were at least as effective as the volumetric measures in predicting memory performance in community-dwelling individuals in the ninth or tenth decade of life. PMID- 16365742 TI - Editorial. PMID- 16365741 TI - HLA class I and class II frequencies in patients with cutaneous malignant melanoma from southeastern Spain: the role of HLA-C in disease prognosis. AB - Available data have led to a controversy on the relationship between human leukocyte antigen (HLA) and cutaneous malignant melanoma susceptibility or prognosis. Moreover, the influence of HLA-C on melanoma has not yet been well established. Therefore, the aim of the current study was to analyze the possible influence of the HLA system on melanoma susceptibility and prognosis in the Spanish population. For this purpose, HLA-A and HLA-B serotyping and HLA-C, HLA DRB1, and HLA-DQB1 genotyping by polymerase chain reactions using sequence specific oligonucleotide (PCR-SSO) and sequence-specific primer (PCR-SSP) were performed in 174 melanoma patients and 227 ethnically matched controls. The number of controls was increased up to 356 for HLA-C typing. Patients were stratified according to the histological subtypes of melanoma, sentinel lymph node status, tumor thickness, and ulceration of primary lesion. No HLA-A, HLA-B, HLA-DRB1, or HLA-DQB1 relationship with melanoma was observed for susceptibility or disease prognosis. However, the analysis of HLA-C locus showed that individuals homozygous for HLA-C(Lys80) were significantly more frequent within the patient than the control group. Remarkably, individuals homozygous for group 2 HLA-C alleles (HLA-C(Lys80)) seem to be associated with metastatic progression of melanoma. In contrast, we found a negative association between group 1 HLA-C alleles (HLA-C(Asn80)) and disease susceptibility or metastasis development. In conclusion, although an association with HLA-A, HLA-B, HLA-DRB1, or HLA-DQB1 was not demonstrated, the study of the HLA-C locus revealed that the analysis of the dimorphism at position 80 in the alpha1 helix may help to evaluate the risk and prognosis of melanoma in our population. PMID- 16365743 TI - [Preoperative radiochemotherapy of advanced resectable cancer of the oral cavity with cisplatin vs paclitaxel/carboplatin. Analysis of two multimodality treatment concepts]. AB - PURPOSE: The purpose of simultaneous chemoradiotherapy is to increase local tumor control and to decrease the incidence of distant metastases. Platinum-based chemotherapy regimens are widely accepted as standard treatment in advanced head and neck cancer. Due to their remarkable activity as well as radiosensitizing effects, taxanes play in increasing role in multimodality treatment concepts of solid tumors. MATERIAL AND METHODS: Since 1989 a total of 85 patients with advanced squamous cell carcinoma of the oral cavity and oropharynx were recruited to participate in two different prospective clinical trials of preoperative chemoradiation. Both treatment concepts (study A: cisplatin 12.5 mg/m(2) plus radiation 36 Gy vs study B: Taxol 40 mg/m(2)/carboplatin AUC 1.5 plus radiation 40 Gy) were compared and further statistical analysis including descriptive analysis as well as univariate and multivariate analysis was performed. RESULTS: At the time of the analysis a total of 47 patients (55.3%) were alive, 38 patients (44.7%) died from tumor-related causes. The overall survival for both patient groups at 5 years was 45.1% (study A) and 84.9% (study B) and after 10 years 32.4% for study group A. A comparative analysis of both groups revealed a highly significant survival benefit at 4 years for study B (p=0.0015). In various calculation models of the multivariate analysis the regimen containing Taxol/carboplatin (study B) was superior to the cisplatin regimen (study A). CONCLUSION: Based on a retrospective analysis of two different concepts of preoperative chemoradiation and supported by recent data from the literature it is justified to conclude that taxane-based combination regimens are more effective compared to cisplatin-based regimens. PMID- 16365744 TI - Magnetic resonance imaging appearance of the flexor carpi radialis tendon after harvest in ligamentous reconstruction tendon interposition arthroplasty. AB - OBJECTIVE: To determine whether the post-harvest magnetic resonance (MR) imaging appearance of flexor carpi radialis (FCR) tendons, harvested during ligamentous reconstruction tendon interposition (LRTI) of the thumb carpometacarpal (CMC) joint arthroplasty, is consistent with tendon regeneration. DESIGN: Operative reports and patient medical records for all patients undergoing LRTI arthroplasty between 1995 and 2003 at our institution were reviewed. MR images of the patients' forearms and wrists were obtained and interpreted by two musculoskeletal radiologists. Using the flexor carpi ulnaris (FCU) tendon as an internal standard, the extent of FCR tendon regeneration was expressed as a percentage by dividing the volume of regenerated FCR tendon by the volume of the FCU tendon. PATIENTS: Fourteen patients who had the full thickness of the FCR tendon harvested and who were available for MR imaging were identified and included in the study. RESULTS AND CONCLUSIONS: At least partial regeneration of the FCR tendon occurred in 11 of the 14 patients (79%). Of these, 2 patients (14%), demonstrated complete, or nearly complete regeneration. Partial regeneration of the FCR tendon was seen in 9 of the 14 patients (64%). In 3 patients (21%), there was no appreciable regeneration of the FCR tendon. Among patients who underwent full-thickness harvest of the FCR tendon for LRTI arthroplasty of the first CMC joint, the follow-up MR imaging appearance of the flexor carpi radialis tendon was consistent with tendon regeneration in 79% of those examined. PMID- 16365745 TI - Estimation of pelvic tilt on anteroposterior X-rays--a comparison of six parameters. AB - OBJECTIVE: To compare six different parameters described in literature for estimation of pelvic tilt on an anteroposterior pelvic radiograph and to create a simple nomogram for tilt correction of prosthetic cup version in total hip arthroplasty. DESIGN: Simultaneous anteroposterior and lateral pelvic radiographs are taken routinely in our institution and were analyzed prospectively. The different parameters (including three distances and three ratios) were measured and compared to the actual pelvic tilt on the lateral radiograph using simple linear regression analysis. PATIENTS: One hundred and four consecutive patients (41 men, 63 women with a mean age of 31.7 years, SD 9.2 years, range 15.7-59.1 years) were studied. RESULTS: The strongest correlation between pelvic tilt and one of the six parameters for both men and women was the distance between the upper border of the symphysis and the sacrococcygeal joint. The correlation coefficient was 0.68 for men (P<0.001) and 0.61 for women (P<0.001). Based on this linear correlation, a nomogram was created that enables fast, tilt-corrected cup version measurements in clinical routine use. CONCLUSION: This simple method for correcting variations in pelvic tilt on plain radiographs can potentially improve the radiologist's ability to diagnose and interpret malformations of the acetabulum (particularly acetabular retroversion and excessive acetabular overcoverage) and post-operative orientation of the prosthetic acetabulum. PMID- 16365746 TI - An unusual variant of intraneural ganglion of the common peroneal nerve. AB - A highly unusual variant of an intraneural ganglion of the common peroneal nerve in a 30-year-old male is presented. There was extrusion of the contents of the cyst into the substance of the nerve, dissecting between the fibres and expanding the nerve in such a way that it mimicked an intraneural tumour clinically, radiologically and histologically. A comprehensive review of the entity is undertaken. PMID- 16365747 TI - Efficacy of diverse high-dose regimens followed by autologous peripheral blood stem cell transplantation in consecutive multiple myeloma patients: a single centre analysis over a 12-year period. PMID- 16365748 TI - The gap between evidence-based medicine and daily practice in the management of paediatric asthma. A pharmacy-based population study from The Netherlands. AB - OBJECTIVE: We evaluated the adherence to national guidelines for the treatment of asthma in childhood. METHODS: Prescriptions for anti-asthma medication for children (0-14 years of age) were retrieved from the InterAction DataBase (IABD) for the year 2002. These were compared with recommendations found in national guidelines. RESULTS: Anti-asthma medication was prescribed for 3,612 children (5%) of the paediatric population. Inhaled medication was prescribed for 3,554 (98%) children. In 1,940 of 1,993 (97%) of the children under the age of 6 years pressurised metered dose inhalers (pMDIs) were given. Short-acting beta2-agonists had not been prescribed in the previous 2-year period in 559 children (15%), 543 children older than 8 years (36%) did not receive a prescription for a dry powder inhalator and 239 children (7%) had more than one type of inhalator. Long-acting beta2-agonists were prescribed in 396 children, but without concomitant inhaled corticosteroids (ICS) in 35 children (9%). CONCLUSIONS: Inhalation therapy as the method of choice in asthma therapy and the use of pMDI in preschool children are widely accepted in the Netherlands. Not all children have been prescribed bronchodilators. Some children have more than one type of inhaler device and others use long-acting beta2-agonists not in combination with ICS. Although national and international guidelines about the treatment of asthma in children offer evidence-based advice, important principles are not followed. Effective interventions aimed at implementing existing guidelines into daily practice are urgently needed. PMID- 16365749 TI - Influence of weather on emergency transport events coded as stroke: population based study in Japan. AB - Studying the relation between incidence of stroke and weather is difficult because it requires large-scale community-based data collection. Despite the lack of strong evidence that weather conditions influence stroke incidence, many clinicians feel that meteorological conditions influence the onset of stroke. This study examined whether emergency events related to stroke are influenced by meteorological factors and was based on computerized records of emergency medical transport services in a Japanese city during the period January 1992-December 2003. A total of 53,585 patients transported for an event coded as stroke were analyzed in relation to meteorological factors such as temperature, humidity, and barometric pressure. Poisson regression analysis was applied to clarify the influence of daily meteorological conditions on the daily incidence of emergency transport due to events coded as stroke. Ordinary least squares regression analysis was used to evaluate the influence of weather, defined as the combination of meteorological parameters, on the occurrence of emergency transport due to events coded as stroke. Daily mean ambient temperature and daily mean relative humidity showed a statistically significant negative effect on the incidence of the emergency transport events for both men and women (P<0.001). Daily mean barometric pressure was not significantly related to these events. The occurrence of a holiday was negatively related to the incidence (P<0.001). Dry weather and cool weather were likely to shift the circadian curve of the incidence upward. Thus, occurrence of emergency transport due to events coded as stroke is likely to be associated with weather conditions. PMID- 16365750 TI - Subclinical effects of prenatal methylmercury exposure on cardiac autonomic function in Japanese children. AB - OBJECTIVES: The subclinical effects of prenatal exposure to methylmercury from fish consumption on the cardiac autonomic function were assessed in 136 Japanese 7-year-old children recruited for this study. METHODS: Samples of child's hair and dry umbilical cord preserved were collected, and hair mercury and cord tissue methylmercury concentrations were determined as current and prenatal exposure biomarkers, respectively. Cardiac autonomic indicators of parasympathetic and sympathetic activities were calculated from the electrocardiographic RR intervals measured. RESULTS: In the children, the cord tissue methylmercury (0.017-0.367, median 0.089 microg/g) was not significantly correlated with the hair mercury (0.43-6.32, median 1.66 microg/g). The cord tissue methylmercury was related negatively to parasympathetic components of cardiac autonomic indicators (P < 0.05) and positively to sympathovagal indices (P < 0.05), even after correction for possible confounders such as age and sex, although the hair mercury was not significantly correlated with any cardiac autonomic indicators. CONCLUSIONS: Despite the potential limitations involved in the retrospective study, these findings suggest that prenatal methylmercury exposure (median of estimated maternal hair mercury at parturition, 2.24 microg/g) may be associated with reduced parasympathetic activity and/or sympathovagal shift. PMID- 16365751 TI - Examination of the influence of personal traits and habituation on the reporting of complaints at experimental exposure to ammonia. AB - OBJECTIVE: The aim of the study is to examine the impact of personal traits and habituation on the intensity of self reported health symptoms and complaints. METHODS: Forty-three healthy male volunteers were exposed to ammonia vapours in concentrations of 0, 10, 20, 20/40, and 50 ppm on five consecutive days. To explore a possible influence of habituation on the perception of symptoms, the study group was divided into 30 men who were not familiar with the smell of ammonia, and ten participants regularly exposed to ammonia at the workplace. The perceived acute health symptoms and self-reported changes of well-being were assessed by the German version of a questionnaire of the Swedish Performance Evaluation System (SPES). Personal traits were ascertained with the positive and negative affectivity schedule (PANAS) and the Freiburger Personlichkeits Inventar (FPI). RESULTS: There are significant associations between personal traits and the amount of complaints. Subjects with high positive affectivity report less respiratory and irritative complaints, whereas persons with negative affectivity report significantly more olfactory and respiratory symptoms. In general, the strength of these correlations decreases with higher exposure levels. At ammonia concentration above 20 ppm, these associations were no more statistically significant. During the daily exposures, the score of symptoms did not vary significantly. The perceived intensity of health symptoms and annoyance increased with the concentration of ammonia, while the self-reported dimensions of well being decreased. Workers familiar with the smell of ammonia vapours reported less symptoms compared to naive subjects. CONCLUSIONS: Habituation to ammonia vapours as well as personal traits influence the reporting of complaints particularly at low exposure. Both factors should be considered in the examination of chemosensory irritative compounds. PMID- 16365752 TI - Somatosensory graviception inhibits soleus H-reflex gain in humans during walking. AB - To investigate the effects of gravity-related somatosensory information on spinal human reflexes, the soleus H-reflex was recorded in ten healthy subjects walking on a treadmill at 2.0 km/h on land and in water. The modulation pattern of the soleus H-reflex was determined in ten different phases of the step cycle. While the subjects were walking in water, the background electromyographic activity (BGA) of the soleus was lower than that on land; on the other hand, the soleus H reflex amplitude while the subjects were walking in water showed no significant differences throughout the step cycle compared with that while the subjects were on land; the phase-dependent soleus H-reflex modulation pattern was well preserved while walking in water. There was a linear relationship between the BGA and the H-reflex amplitude in each condition; however, the soleus H-reflex gain while walking in water was significantly higher than that on land. These findings suggest that the somatosensory graviception can markedly reduce the spinal reflex excitability. Our findings are discussed in relation to human gait; therefore, further studies are needed to clarify the effect of somatosensory graviception on human neural mechanisms. PMID- 16365753 TI - Methylenetetrahydrofolate reductase polymorphism and minor increase of risk for oral cancer. AB - PURPOSE: We investigated whether the mutant methylenetetrahydrofolate reductase (MTHFR) increases risk for oral cancer. The common germ-line mutation C677T in the MTHFR gene significantly diminishes specific activity of the enzyme, which is responsible for the circulating form of folate. Folate deficiency is associated with increased risk for thrombosis, as well as for several types of cancer, through disruption of DNA methylation, DNA synthesis and deficient DNA repair. METHODS: We searched for the C677T mutation by restriction fragment analysis of PCR products in DNA samples of 110 patients with oral squamous cell carcinoma and 120 healthy controls of comparable ethnicity, age and sex. RESULTS: The number of heterozygotes was significantly different in the two groups (P<0.005), as well as in subgroups of patients with or without a positive family history for cancer, compared to normal controls (P<0.01 and P<0.005, respectively). Furthermore, the subgroup of patients with a positive family history for thrombophilia had a significant increase both in the frequencies of mutant alleles (P<0.01) and heterozygotes (P<0.001) in comparison to normal controls. CONCLUSIONS: The obtained results suggest that the MTHFR mutation is a minor contributing factor in oncogenesis in the oral region, in conjunction with low dietary uptake of folate. PMID- 16365754 TI - Autonomic dysfunction in systemic lupus erythematosus. AB - The objectives were to study the frequency and pattern of autonomic dysfunction in systemic lupus erythematosus (SLE). Fifty-one patients of SLE and 30 age- and sex-matched healthy controls were studied prospectively using a standard battery of noninvasive tests. Autonomic symptoms were seen in 37% patients. On laboratory testing incipient dysfunction was seen in 9 (18%) cases and 1 (3%) control, while atypical involvement was seen in 11 (21%) cases and 6 (20%) controls. Autonomic dysfunction did not correlate with disease duration, lupus activity, disease damage, any particular organ involvement or the presence/absence of peripheral neuropathy. Autonomic neuropathy is not uncommon in lupus and may exist independent of peripheral neuropathy. There are no specific clinical predictors. The clinical significance of autonomic dysfunction detected by laboratory testing warrants longitudinal studies. PMID- 16365755 TI - Macrophage-like synoviocytes display phenotypic polymorphisms in a serum-free tissue-culture medium. AB - Synovial macrophages play an outstanding role in many rheumatic diseases. However, traditional serum-containing tissue-culture techniques hamper in vitro studies due to fibroblast activation not found in vivo. The objective of this study was to examine dissociated synovial cells in a macrophage-selective, serum free tissue-culture medium. Osteoarthritis synovial tissue (n=11) was cultured in Iscove's Modified Dulbecco's Medium (IMDM) with 10% fetal bovine serum (FBS) and compared to a serum-free, insulin-supplemented medium. After 9-11 and 19-21 days in vitro, immunohistochemistry was performed for macrophage/lymphocyte markers and cell division. Cytokine profiles were determined by RT-PCR. In serum, cells with a bipolar morphology rapidly proliferated. Respectively, 14.34+/-12.94% and 13.25+/-12.66% expressed CD68 and HLA-DR. These markers further decreased after one passage. In serum-free medium, proliferation was infrequent, and cells with diverse morphologies expressed 83.10+/-6.80% and 55.03+/-6.88% CD68 and HLA-DR respectively. CD14 was rare, and lymphocytes were missing. Both cultures expressed interleukin-6 and interleukin-8. This novel serum-free method permits the culture of distinct CD68/HLA-DR associated phenotypes. PMID- 16365756 TI - An integrated map of Oryza sativa L. chromosome 5. AB - The developments of molecular marker-based genetic linkage maps are now routine. Physical maps based on contigs of large insert genomic clones have been established in several plant species. However, integration of genetic, physical, and cytological maps is still a challenge for most plant species. Here we present an integrated map of rice (Oryza sativa L.) chromosome 5, developed by fluorescence in situ hybridization mapping of 18 bacterial artificial chromosome (BAC) clones or PI-derived artificial chromosome (PAC) clones on meiotic pachytene chromosomes. Each BAC/PAC clone was anchored by a restriction fragment length polymorphism marker mapped to the rice genetic linkage map. This molecular cytogenetic map shows the genetic recombination and sequence information of a physical map, correlated to the cytological features of rice chromosome 5. Detailed comparisons of the distances between markers on genetic, cytological, and physical maps, revealed the distributions of recombination events and molecular organization of the chromosomal features of rice chromosome 5 at the pachytene stage. Discordance of distances between the markers was found among the different maps. Our results revealed that neither the recombination events nor the degree of chromatin condensation were evenly distributed along the entire length of chromosome 5. Detailed comparisons of the correlative positions of markers on the genetic, cytological, and physical maps of rice chromosome 5 provide insight into the molecular architecture of rice chromosome 5, in relation to its cytological features and recombination events on the genetic map. The prospective applications of such an integrated cytogenetic map are discussed. PMID- 16365757 TI - Isolation of S-locus F-box alleles in Prunus avium and their application in a novel method to determine self-incompatibility genotype. AB - This study characterises a series of 12 S-locus haplotype-specific F-box protein genes (SFB) in cherry (Prunus avium) that are likely candidates for the pollen component of gametophytic self-incompatibility in this species. Primers were designed to amplify 12 SFB alleles,including the introns present in the 50 untranslated region;sequences representing the S-alleles S1, S2, S3, S4, S40, S5,S6, S7, S10, S12, S13 and S16 were cloned and characterized. [The nucleotide sequences reported in this paper have been submitted to the EMBL/GenBank database under the following accession numbers: PaSFB1(AY805048), PaSFB2 (AY805049), PaSFB3 (AY805057),PaSFB4 (AY649872), PaSFB40 (AY649873), PaSFB5(AY805050), PaSFB6 (AY805051), PaSFB7 (AY805052),PaSFB10 (AY805053), PaSFB12 (AY805054), PaSFB13(AY805055), PaSFB16 (AY805056).] Though the coding regions of six of these alleles have been reported previously,the intron sequence has previously been reported only for S6. Analysis of the introns revealed sequence and length polymorphisms. A novel, PCR-based method to genotype cultivars and wild accessions was developed which combines fluorescently labelled primers amplifying the intron of SFB with similar primers for the first intron of S-RNase alleles. Intron length polymorphisms were then ascertained using a semi-automated sequencer. The convenience and reliability of this method for the determination of the self-incompatibility (SI) genotype was demonstrated both in sweet cherry cultivars representing alleles S1 to S16 and in individuals from a wild population encompassing S-alleles S17 to S22. This method will greatly expedite SI characterisation in sweet cherry and also facilitate large-scale studies of self-incompatibility in wild cherry and other Prunus populations. PMID- 16365758 TI - Mapping of barley homologs to genes that regulate low temperature tolerance in Arabidopsis. AB - We investigated the allelic nature and map locations of Hordeum vulgare (barley) homologs to three classes of Arabidopsis low temperature (LT) regulatory genes CBFs, ICE1, and ZAT12-to determine if there were any candidates for winterhardiness-related quantitative trait loci (QTL). We phenotyped the Dicktoo x Morex (DxM) mapping population under controlled freezing conditions and in addition to the previously reported 5H-L Fr-H1 QTL, observed three additional LT tolerance QTLs on 1H-L, 4H-S, and 4H-L. We identified and assigned either linkage map or chromosome locations to 1 ICE1 homolog, 2 ZAT12 homologs, and 17 of 20 CBF homologs. Twelve of the CBF genes were located on 5H-L and the 11 with assigned linkage map positions formed 2 tandem clusters on 5H-L. A subset of these CBF genes was confirmed to be physically linked, validating the map position clustering. The tandem CBF clusters are not candidates for the DxM LT tolerance Fr-H1 QTL, as they are approximately 30 cM distal to the QTL peak. No LT tolerance QTL was detected in conjunction with the CBF gene clusters in Dicktoo x Morex. However, comparative mapping using common markers and BIN positions established the CBF clusters are coincident with reported Triticeae LT tolerance and COR gene accumulation QTLs and suggest one or more of the CBF genes may be candidates for Fr-H2 in some germplasm combinations. These results suggest members of the CBF gene family may function as components of winter-hardiness in the Triticeae and underscore both the importance of extending results from model systems to economically important crop species and in viewing QTL mapping results in the context of multiple germplasm combinations. PMID- 16365759 TI - Comparison of phenotypic and molecular distances to predict heterosis and F1 performance in Ethiopian mustard (Brassica carinata A. Braun). AB - Predicting heterosis and F1 performance from the parental generation could largely enhance the efficiency of breeding hybrid or synthetic cultivars. This study was undertaken to determine the relationship between parental distances estimated from phenotypic traits or molecular markers with heterosis, F1 performance and general combining ability (GCA) in Ethiopian mustard (Brassica carinata). Nine inbred lines representing seven different geographic regions of Ethiopia were crossed in half-diallel. The nine parents along with their 36 F1s were evaluated in a replicated field trail at three locations in Ethiopia. Distances among the parents were calculated from 14 phenotypic traits (Euclidean distance, ED) and 182 random amplified polymorphic DNA (RAPD) markers (Jaccard's distances, JD), and correlated with heterosis, F1 performance and GCA sum of parents (GCAsum). The correlation between phenotypic and molecular distances was low (r=0.34, P< or =0.05). Parents with low molecular distance also had low phenotypic distance, but parents with high molecular distance had either high, intermediate or low phenotypic distance. Phenotypic distance was highly significantly correlated with mid-parent heterosis (r=0.53), F1 performance (r=0.61) and GCA (r=0.79) for seed yield. Phenotypic distance was also positively correlated with (1) heterosis, F1 performance and GCA for plant height and seeds plant(-1), (2) heterosis for number of pods plant(-1), and (3) F1 performance for 1,000 seed weight. Molecular distance was correlated with GCAsum (r=0.36, P< or =0.05) but not significantly with heterosis and F1 performance for seed yield. For each parent a mean distance was calculated by averaging the distances to the eight other parents. Likewise, mean heterosis was estimated by averaging the heterosis obtained when each parent is crossed with the other eight. For seed yield, both mean ED and JD were significantly correlated with GCA (r=0.90, P< or =0.01 for ED and r=0.68, P< or =0.05 for JD) and mean heterosis (r=0.79, P< or =0.05 for ED and r=0.77, P< or =0.05 for JD). In conclusion, parental distances estimated from phenotypic traits better predicted heterosis, F1 performance and GCA than distances estimated from RAPD markers. PMID- 16365761 TI - Quantitative trait loci analysis for the developmental behavior of Soybean (Glycine max L. Merr.). AB - Quantitative trait loci (QTLs) identified so far in soybean were mainly derived in the final stage of plant development, which did not apply to the exploitation of genetic effects that were expressed during a specific developmental stage. Thus, the aim of this study was to identify conditional QTLs associated with yield traits at a specific developmental interval of soybean plant. The 143 recombinant inbred lines developed from the cross of soybean cultivars 'Charleston' and 'Dongnong 594' were used for the developmental QTLs analysis of pod number in the main stem and plant height by composite interval mapping method combined with mixed genetic model. The results indicated that the number and type of QTLs and their genetic effects for the two agronomic traits were different in a series of measuring stages. A total of 10 unconditional QTLs in 6 linkage groups and 5 conditional QTLs in 3 linkage groups were identified for the pod number of the main stem, while 13 unconditional QTLs in 7 linkage groups and 12 conditional QTLs in 6 linkage groups were identified for plant height. Many QTLs that were detected in the early stages were different from those detected at the later stages. Some QTLs existed only at one stage and others existed across two or three stages. Five marker intervals (satt509-satt251, sat_099-sat_113, sat_113 OPAW19_4, satt457-OPC10_85, sat_095-OPBA08_5) were proven to be associated both with the development of pod number in the main stem and the development of plant height. The present study suggested that the development of pods and plant height in soybean were governed by time-dependent gene expression. PMID- 16365760 TI - A major gene mapped on chromosome XII is the main factor of a quantitatively inherited resistance to Meloidogyne fallax in Solanum sparsipilum. AB - Meloidogyne fallax is an emerging pest in Europe and represents a threat for potato production. We report the mapping of genetic factors controlling a quantitative resistance against M. fallax identified in the Solanum sparsipilum genotype 88S.329.15. When infected, this genotype develops a necrotic reaction at the feeding site of the juveniles and totally prevents their development to the female stage. A "F1" diploid progeny consisting of 128 individuals was obtained using the potato (S. tuberosum) dihaploid genotype BF15 H1 as female progenitor. Sixty-eight hybrid genotypes displayed necrosis at the feeding site of the juveniles and 60 other genotypes showed no defence reaction. This suggested a monogenic control of the resistance. However, when considering the number of nematode females developed in their roots, a continuous distribution was observed for both "necrotic" and "non-necrotic" hybrid genotypes, indicating a polygenic control of the resistance. A linkage map of each parental genotype was constructed using AFLP markers. The necrotic reaction (NR) was mapped as a qualitative trait on chromosome XII of the resistant genotype 88S.329.15. Quantitative trait locus (QTL) analysis for the number of nematode females developed per "F1" plant genotype was performed using the QTL cartographer software. No QTL was detected on the linkage map of the susceptible parent. A QTL explaining 94.5% of the phenotypic variation was mapped on chromosome XII of the resistant progenitor. This QTL, named MfaXIIspl, was mapped in a genomic region collinear to the map position of the Mi-3 gene conferring resistance to Meloidogyne incognita in tomato. It corresponds to the NR locus. PMID- 16365762 TI - [Methods of burn treatment. Part I: general aspects]. AB - Burns and scalds are common injuries that present with a wide range of severity. Correct evaluation of a burn's depth and extent is essential for adequate treatment, not only initially but also for late results. The depth of a burn is classified as first-to-third degree, and its extent can be deducted from specific tables. As a generalised haemodynamic reaction, a capillary leak allows fluid and colloidal substances to leave the intravasal system. This can lead to hypovolemic shock. In the first 24 h, only cristalloid fluid according to Baxter's formula should be administered. Transfer to a burn centre is indicated in accordance with well-defined guidelines. Concomitant injuries, especially inhalation traumata, need to be diagnosed and treated early. By activation of the immune response, a sepsis-like immune response syndrome can occur,resulting in bacterial translocation and colonisation with high mortality rates. PMID- 16365763 TI - [Genetics and pathophysiology of obesity]. AB - Obesity is one of the major health problems in developed countries. More than 35% of adults in Germany are considered to be overweight (BMI 25-29.9 kg/m(2)) and approximately 15-20% to be obese (BMI >30 kg/m(2)). Overweight and obesity result from chronic disruption of energy balance. Recent progress in understanding of appetite control and energy expenditure has elucidated a complex integrated system of energy homoeostasis. Although changes in nutritional habits and reduction of physical activity are the main characteristics leading to the strong increase in obesity, it is now recognized that obesity is not only caused by a lack of will power, but can be a consequence of metabolic defects. As obesity results from a genetic make-up favoring weight gain in an "obesigenic" environment the elucidation of the underlying molecular mechanisms might be translated in novel therapeutic options in the future. PMID- 16365764 TI - [Check-up-examinations in internal medicine]. AB - A growing body of evidence supports or rejects preventive interventions in asymptomatic adults. Thus, counseling for smoking cessation and some immunizations have been shown to be cost-effective. Evidence supports screening for body weight (obesity), hypertension, hyperlipidemia, cervical cancer, colorectal cancer and breast cancer. Screening for lung, pancreatic and ovarian cancer has no effect on outcome and should not be performed. Controversial preventive interventions include general screening for diabetes mellitus in the young adult, thyroid disorders and prostate cancer. Physicians should be aware of a possible hidden agenda in patients presenting for a check-up. PMID- 16365765 TI - Propensity for paternal inheritance of de novo mutations in Alexander disease. AB - De novo dominant mutations in the GFAP gene have recently been associated with nearly all cases of Alexander disease, a rare but devastating neurological disorder. These heterozygous mutations must occur very early in development and be present in nearly all cells in order to be detected by the sequencing methods used. To investigate whether the mutations may have arisen in the parental germ lines, we determined the parental chromosome bearing the mutations for 28 independent Alexander disease cases. These cases included 17 different missense mutations and one insertion mutation. To enable assignment of the chromosomal origin of the mutations, six new single nucleotide polymorphisms in the GFAP gene were identified, bringing the known total to 26. In 24 of the 28 cases analyzed, the paternal chromosome carried the GFAP mutation (P < 0.001), suggesting that they predominantly arose in the parental germ line, with most occurring during spermatogenesis. No effect of paternal age was observed. There has been considerable debate about the magnitude of the male to female germ line mutation rate; our ratio of 6:1 is consistent with indirect estimates based on the rate of evolution of the sex chromosome relative to the autosomic chromosomes. PMID- 16365766 TI - Phage display systems and their applications. AB - Screening phage display libraries of proteins and peptides has, for almost two decades, proven to be a powerful technology for selecting polypeptides with desired biological and physicochemical properties from huge molecular libraries. The scope of phage display applications continues to expand. Recent applications and technical improvements driving further developments in the field of phage display are discussed. PMID- 16365767 TI - Length-dependent deactivation of ventricular trabeculae in the bivalve, Spisula solidissima. AB - Shortening-deactivation has been identified and characterized in ventricular trabeculae of the bivalve, Spisula solidissima (Heterodonta, Mactridae). This muscle had ultrastructural similarities to vertebrate smooth muscle. Deactivation was defined as the fraction of maximal force lost during a contraction when a muscle is shortened rapidly (by a quick-release, QR) to a known length, relative to a control isometric contraction at that same length. The magnitude of deactivation was dependent on the size of the release and the point at which the release was applied during the cycle of contraction. QR/quick-stretch (QS) perturbations at the same point during the contraction resulted in negligible deactivation. The magnitude of deactivation was independent of shortening rate. Deactivation was attenuated by applying caffeine (100 microM) and blocked with high extracellular Ca(2+) (56 mM). The Ca(2+) ionophore, A23187 (10 microM), augmented deactivation as did the positive inotrope serotonin (100 nM). Treatment with ryanodine (5 microM) had no significant effect on deactivation. These results suggest that a reduction in Ca(2+) at the contractile element and/or sequestration of Ca(2+) may occur during shortening. Deactivation may minimize the magnitude of work done during active shortening of bivalve cardiac muscle, particularly against the low afterload exhibited in the bivalve peripheral circulatory system. Intracellular Ca(2+) fluxes during sudden length perturbations may explain the effect of stretch on action potential duration in the bivalve heart, as shown previously. PMID- 16365768 TI - [Observations of normal and ACL-deficient knee joints after stress MRI]. AB - BACKGROUND: The Lachman test is the most reliable clinical test for diagnosing rupture of the anterior cruciate ligament (ACL). Previous X-ray studies have presented a "radiologic Lachman test". Recently anterior tibial translation was demonstrated using open access MRI. Two methods were developed to transfer a similar technique to a more widely available closed MRI. METHODS: Using closed MRI we investigated 22 knees in 21 patients with pure rupture of the ACL. Anteriorly and posteriorly directed shear forces were applied to the tibiofemoral joint at 20 degrees flexion either by positioning a 9-kg load on the distal femur (method 1) or performing a semi-manual Lachman test with a custom-made orthosis (method 2). RESULTS: Both methods produced relative anterior tibial translation in both compartments of the normal and ACL-deficient knee which could be measured on sagittal images. They were greater laterally than medially and in injured than in uninjured knees. However, instability of the medial compartment predicted clinical and symptomatic instability as translation was posterior to positions achieved in normal knees during the active and passive flexion arc. CONCLUSION: A Lachman sign can be produced in a closed magnet with different methods and findings can be used for more precise information regarding kinematics and degree of instability and could be helpful if surgical treatment is necessary. PMID- 16365769 TI - Complex memories in honeybees: can there be more than two? AB - Foraging honeybees are likely to learn visual and chemical cues associated with many different food sources. Here, we explore how many such sources can be memorized and recalled. Marked bees were trained to visit two (or three) sugar feeders, each placed at a different outdoor location and carrying a different scent. We then tested the ability of the bees to recall these locations and fly to them, when the training scents were blown into the hive, and the scents and food at the feeders were removed. When trained on two feeder locations, each associated with a different scent, the bees could correctly recall the location associated with each scent. However, this ability broke down when the number of scents and feeder locations was increased to three. Performance was partially restored when each of the three training feeders was endowed with an additional cue, namely, a distinct colour. Our results suggest that bees can recall a maximum of two locations when each is associated with a different scent. However, this number can be increased if the scent cues are augmented by visual cues. These findings have implications for the ways in which associations are established and laid down in honeybee memory. PMID- 16365770 TI - A molecular-dynamics simulation study of diffusion of a single model carbonic chain on a graphite (001) surface. AB - Molecular-dynamics simulations have been used to study the diffusion of a short single model carbonic chain on the graphite (001) surface. The calculated diffusion coefficient (D) first increases, then decreases with increasing chain length (N). This abnormal behavior is similar to polymer lateral diffusion at the solid-liquid interface. Furthermore, we have studied the relation between the mean-square gyration radius and N. [Figure: see text]. PMID- 16365771 TI - Mesoscopic simulation study on a weakly charged block polyelectrolyte in aqueous solution. AB - In this paper, we use density functional theory to study the effect of the charge of solvophilic beads and concentration on the mesoscale structures of polyelectrolyte solution. The polyelectrolyte A6B12A6 was selected as the triblock polymer, and the solvophobic B blocks have no charges, while the solvophilic A blocks are charged. The simulation results showed: at higher concentration (above 50% systems), relatively small charges on the solvophilic block do not alter the bicontinuous phase inherent to uncharged solution, but at moderate concentrations (50% system), even though the charge per solvophilic bead is very small, the order lamellar structures become disturbed. [Figure: see text]. PMID- 16365772 TI - Prediction of beta-strand packing interactions using the signature product. AB - The prediction of beta-sheet topology requires the consideration of long-range interactions between beta-strands that are not necessarily consecutive in sequence. Since these interactions are difficult to simulate using ab initio methods, we propose a supplementary method able to assign beta-sheet topology using only sequence information. We envision using the results of our method to reduce the three-dimensional search space of ab initio methods. Our method is based on the signature molecular descriptor, which has been used previously to predict protein-protein interactions successfully, and to develop quantitative structure-activity relationships for small organic drugs and peptide inhibitors. Here, we show how the signature descriptor can be used in a Support Vector Machine to predict whether or not two beta-strands will pack adjacently within a protein. We then show how these predictions can be used to order beta-strands within beta-sheets. Using the entire PDB database with ten-fold cross-validation, we have achieved 74.0% accuracy in packing prediction and 75.6% accuracy in the prediction of edge strands. For the case of beta-strand ordering, we are able to predict the correct ordering accurately for 51.3% of the beta-sheets. Furthermore, using a simple confidence metric, we can determine those sheets for which accurate predictions can be obtained. For the top 25% highest confidence predictions, we are able to achieve 95.7% accuracy in beta-strand ordering. [Figure: see text]. PMID- 16365773 TI - A new shoulder orthosis for paralysis of the trapezius muscle after radical neck dissection: a preliminary report. AB - Despite recent advances using more conservative approaches, standard classical radical neck dissection is still one of the most frequently performed procedures in head and neck cancer patients who have advanced metastatic neck disease. The trapezius muscle paralysis following division of the spinal accessory nerve results in severe pain and cosmetic disturbance related to malalignment of the shoulder joint. The objective of this study is to report our results with a newly developed orthosis to prevent and correct shoulder dysfunction following standard radical neck dissection. Thirty-four patients who underwent standard radical neck dissection as a part of their surgical treatment from 1997-2002 were rehabilitated by the shoulder orthosis. Beginning 2 weeks after surgery, the patients were allowed to use their orthosis. By using a standard questionnaire, the pain and activity scores were recorded at the 1st, 3rd, 6th and 12th months postoperatively. Six patients were excluded from the study, of whom two succumbed to their disease and four discontinued the use of the orthosis. Of 28 patients included in the study, 20 (72%) were completely pain free within 3 months following the surgery. Four patients (14%) noted their pain level as tolerable, and four patients (14%) reported no considerable gain in the pain threshold and/or physical activity levels. Despite the fact that the active abduction range increased only 5 to 20 degrees , the relief of pain and improved malalignment of the scapula and consequently clavicle and humerus led to functional gains, which increased the patients' endurance. At the end of the study, 23 patients (82%) were able to return to their previous jobs or activity levels. Current preliminary reports suggest that this orthosis can be recommended to prevent significant disability in patients with trapezius palsy due to ablative cancer surgery or other reasons. PMID- 16365774 TI - [Systemic therapy of chronic venous diseases]. AB - The therapy of chronic venous insufficiency is multifactorial. Compression, interventional and operative approaches are available along with the possibility of systemic treatment. The efficacy of systemic venotonic medications, mostly phytotherapeutic agents, is controversial. Nonetheless in a number of clinical and laboratory studies, an effect was seen after use for 8-12 weeks. When administered appropriately, venotonic agents can show anti-edematous, anti inflammatory, anti-oxidative, proteolytic effects as well as reducing capillary leakage. Furthermore they increase vein tone and lymph flow. Venotonic agents should be used if compression therapy alone is either not sufficient, contraindicated or not tolerable. They can be useful as a temporizing measure until surgical intervention is performed. Some of them can even be used in pregnancy, but the indications are very strict. PMID- 16365775 TI - [Fetal lung development on MRT. Normal course and impairment due to premature rupture of membranes]. AB - A well-organized interplay between many molecular factors as well as mechanical forces influence fetal lung development. At the end of this complex process a sufficiently sized and structurally mature organ should ensure the postnatal survival of the newborn. Besides prenatal ultrasonography, magnetic resonance imaging (MRI) can now be used to investigate normal and pathological human lung growth in utero. Oligohydramnios, due to premature rupture of membranes (PROM), is an important risk factor for compromised fetal lung growth. In these situations MR volumetry can be used to measure the size of the fetal lung quite accurately. Together with the evaluation of lung signal intensities on T2 weighted sequences, fetuses with pulmonary hypoplasia can be readily detected. PMID- 16365776 TI - [Fetal MRI of pathological brain development]. AB - Because of the superior tissue contrast, high spatial resolution, and multiplanar capabilities, fetal magnetic resonance imaging (MRI) can depict fetal brain pathologies with high accuracy. Pathological fetal brain development may result from malformations or acquired conditions. Differentiation of these etiologies is important with respect to managing the actual pregnancy or counseling future pregnancies. As a widened ventricular system is a common hallmark of both maldevelopment and acquired conditions, it may cause problems in the differential diagnosis. Fetal MRI can provide detailed morphological information, which allows refinement of the diagnosis of ventricular enlargement in a large number of cases. Systematic work-up of morphological details that may be recognized on MR images provides an approach for achieving a correct diagnosis in cases of ventricle enlargement. PMID- 16365777 TI - UV-vis spectroscopic study directly detecting inorganic phosphorus in urine and our reagent kit. AB - The determination of inorganic phosphorus in human urine is very important, since it has diagnostic value in some clinical cases. Here we apply a simple, sensitive and direct method to determine inorganic phosphorus in urine. This new ensemble is prepared by adding ytterbium chloride and pyrocatechol violet in a 2:1 molar ratio in an aqueous solution of 10 mM 2-[4-(2-hydroxyethyl)-1 piperazinyl]ethanesulfonic acid buffer at pH 7.0. The addition of the urine sample turned the blue ensemble yellow and altered the UV-vis absorption spectra. The ensemble exhibits excellent selectivity for inorganic phosphorus over other constituents of urine. We validate the accuracy of our method by the standard procedure (molybdenum blue assay for phosphate). The detection results are basically consistent with normal excretion of phosphate. Furthermore, we fabricated a new kind of inorganic phosphorus reagent kit, which enables us to inspect phosphate concentrations of urine with the naked eye. Fit for all kinds of various clinic uses, our reagent kit is a hopeful substitute for the molybdenum reagent kit. PMID- 16365778 TI - Determination of pharmaceuticals in environmental waters by liquid chromatography/electrospray ionization/tandem mass spectrometry. AB - A high-performance liquid chromatography/electrospray ionization/tandem mass spectrometry (LC/MS-MS) method was developed, validated and used to characterize pharmaceutical inputs in the Grand River watershed, Ontario, Canada. Twenty-seven antibiotics and neutral pharmaceuticals were extracted from aqueous environmental samples in a single step using 13C6-labeled sulfamethazine phenyl as a method surrogate and analyzed by LC/MS-MS. Method detection limits were in the range of 20-1,400 ng/L for the compounds of interest with recoveries from 51 to 130%. Using this method, pharmaceutical profiles in tributaries flowing into the Grand River were investigated using samples collected from seven agricultural sites and one urban site. Quality control data collected during the 5-month field study period showed good method performance. Concentrations of pharmaceuticals in source water with heavy agricultural input and surface waters downstream with urban inputs were determined to examine the effect of agricultural and urban inputs to surface water quality. Only human prescription drugs were found from urban site samples taken at both high-flow (spring and fall) and low-flow (summer) events in 2003, indicating minimal agricultural input. We also found for the first time the presence of the ionophore monensin in the environment as well as the occurrence of the human prescription drug carbamazepine in surface waters receiving primarily agricultural inputs. Possible causes and analytical solutions for some poor recoveries are discussed with respect to extraction pH, total organic carbon and metal ions in the water samples. The ability to extract 27 pharmaceuticals in a single solid-phase extraction step from diverse environmental matrices such as agricultural tile drain, tributary and surface water samples, followed by a single LC/MS-MS analysis, enabled the effective delivery of quality data. PMID- 16365779 TI - Optimizing the analytical performance and construction of ion-selective electrodes with conducting polymer-based ion-to-electron transducers. AB - All-solid-state ion-selective electrodes that use a conducting polymer as the ion to-electron transducer have emerged as one of the most promising classes of all solid-state potentiometric sensors in recent years. This is largely because it has many analytical advantages, including high response stability, which is unique in the field of internal-solution-free ion-selective electrodes. This paper reviews the considerable progress that has been made in this area of sensing in recent years, in terms of detection limits, selectivity coefficients and novel construction methods. PMID- 16365780 TI - Interaction of the mitotic kinesin Eg5 inhibitor monastrol with P-glycoprotein. AB - Monastrol is the first characterised small molecule inhibitor of the motor protein Eg5 involved in bipolar mitotic spindle assembly. Eg5 localises to microtubules in mitosis, but not to interphase microtubules, suggesting that Eg5 inhibitors may be useful to specifically target proliferating tumour tissue, thereby avoiding dose-limiting neuropathy observed with other antimicrotubule agents like taxanes or vinca alkaloids. Because other antimicrotubule agents fail in multidrug resistance associated with P-glycoprotein (Pgp) over-expression, we investigated the interaction of monastrol with Pgp in vitro. By means of the calcein assay (with P388/dx cells and primary porcine brain capillary endothelial cells) and confocal laser-scanning microscopy (with L-MDR1 cells) we demonstrated that monastrol is a weak inhibitor of Pgp in vitro, with f2 values being about two orders of magnitude greater than those of the well-known inhibitors verapamil and quinidine. Monastrol also induces Pgp in vitro as measured by mRNA expression in LS180 cells after incubation with monastrol. However, its effect is weak compared to rifampicin. Whilst it reveals weak inhibitory and inductive characteristics, monastrol appears to be not transported by Pgp, as indicated by the lack of difference in the antiproliferative effect of this compound in cell lines with and without over-expression of Pgp. The observed interaction profile of monastrol with Pgp is promising for the development of other more potent Eg5 inhibitors. PMID- 16365781 TI - Concerning the paper by Mall et al., entitled 'Temperature-based death time estimation with only partially environment conditions' (Int J Legal Med (2005) 119:185-194). PMID- 16365782 TI - Contralateral muscle fatigue in human quadriceps muscle: evidence for a centrally mediated fatigue response and cross-over effect. AB - The purpose of this investigation was to examine the effects of voluntary muscular fatigue in one lower limb and determine whether a 'cross-over' of fatigue is evident in the contralateral limb. Twenty-eight subjects (13 males and 15 females) performed a series of voluntary and evoked isometric contractions of both the dominant (exercised) and non-dominant (non-exercised) leg extensor muscles, prior to and after a fatigue protocol consisting of a 100-s sustained maximal isometric contraction (MVC) performed by the dominant limb only. Force values and surface electromyography (EMG) from the vastus lateralis muscle were obtained allowing for the determination of twitch and compound action potential (M-wave) values. Maximal twitch tension and peak-to-peak amplitude were significantly decreased after the fatigue test in the dominant limb, as was maximal voluntary force (approximately 65 N reduction), EMG activity (approximately 0.1 mV decrease) and voluntary activation (approximately 17% decline). However, no significant changes were observed in the non-dominant limb with respect to twitch and M-wave properties nor in MVC force. The voluntary activation of the non-dominant limb decreased significantly by 8.7% after the fatigue test, which was performed only on the dominant limb. The results of the present study suggest that the decrease in force production in the exercised limb was primarily related to peripheral fatigue mechanisms, with central fatigue making a lesser contribution. Centrally mediated mechanisms appear to be the sole contributor to fatigue in the non-exercised limb suggesting an anticipatory fatigue response and a 'cross-over' of central fatigue between the exercised and non-exercised contralateral limb. PMID- 16365783 TI - Feature binding and episodic retrieval in blindness for congruent stimuli: evidence from analyses of sequential congruency. AB - Targets are identified more poorly when presented during a congruent cued response than during an incongruent cued response (blindness effect). The authors investigated sequential trial dependencies in the blindness effect. The results show that the size of the blindness effect depends both on the previous cued response-target congruency relationship and on repetition of events from the preceding trial. This finding suggests that cued responses and targets become linked together in a single episodic trace; repeating one of these events from the preceding trial activates the other. Depending on whether the activated representation matches or conflicts with events on the current trial, target identification performance is either facilitated or impaired. Implications for action planning and feature binding are discussed. PMID- 16365784 TI - The role of back muscle endurance, maximum force, balance and trunk rotation control regarding lifting capacity. AB - Evaluation of lifting capacity is widely used as a reliable instrument in order to evaluate maximal and safe lifting capacity. This is of importance in regard to planning rehabilitation programs and determining working ability. The aim of this study was to investigate the influence of basic functions on the lifting capacity measured by the progressive isoinertial lifting evaluation (PILE) and the functional capacity evaluation (FCE) tests in a lower (floor to waist) and an upper (waist to shoulder) setting and compare the two test constructs. Seventy four female subjects without acute low back pain underwent an examination of their lifting capacities and the following basic functions: (1) strength and endurance of trunk muscles, (2) cardiovascular endurance, (3) trunk mobility and (4) coordination ability. A linear regression model was used to predict lifting capacity by means of the above-mentioned basic functions, where the F statistics of the variables had to be significant at the 0.05 level to remain in the model. Maximal force in flexion showed significant influence on the lifting capacity in both the PILE and the FCE in the lower, as well as in the upper, lifting task. Furthermore, there was a significant influence of cardiovascular endurance on the lower PILE and also of endurance in trunk flexion on the lower FCE. Additional inclusion of individual factors (age, height, weight, body mass index) into the regression model showed a highly significant association between body height and all lifting tasks. The r (2) of the original model used was 0.19/0.18 in the lower/upper FCE and 0.35/0.26 in the lower/upper PILE. The model r (2) increased after inclusion of these individual factors to between 0.3 and 0.4. The fact that only a limited part of the variance in the lifting capacities can be explained by the basic functions analyzed in this study confirms the assumption that factors not related to the basic functions studied, such as lifting technique and motor control, may have a strong influence on lifting capacity. These results give evidence to suggest the inclusion of an evaluation of lifting capacity in clinical practice. Furthermore, they raise questions about the predictive value of strength and endurance tests in regard to lifting capacity and work ability. PMID- 16365785 TI - Are changes in pain induced by myofeedback training related to changes in muscle activation patterns in patients with work-related myalgia? AB - The objective of this explorative study was to investigate to what extent changes in perceived pain, induced by myofeedback training, are correlated to changes in muscle activation patterns. Thirty subjects with work-related myalgia received myofeedback training. Before (T(0)), directly after (T(1)) and 4 weeks or, in a subset of patients, 3 months after (T(2)) this training, surface electromyography (sEMG) measurements of the upper trapezius muscle were performed during standardized computer tasks; a typing and a stress task. Besides this, visual analogue scales (VAS) were filled in to assess the levels of pain in the neck and shoulders. From the sEMG, root mean square (RMS) and relative rest time (RRT, i.e. the percentage of time RMS is below a certain threshold) were used for data analysis. The relationships between RRT, RMS and VAS at T(0) as well as for the changes between T(1)-T(0) and T(2)-T(0) were investigated using Spearman correlation coefficients. The results revealed no significant correlations between VAS and RMS both at baseline (range R = -0.22 to 0.17) and for the observed changes (range R = -0.33 to 0.32). Also, for VAS and RRT, low correlations were found for baseline (range R = -0.27 to 0.21) and for changes between T(1)-T(0) (range R = -0.02 to 0.38). However, for the changes between T(2)-T(0), correlation coefficients for the VAS for the shoulder and the RRT of the right trapezius during both the typing and stress tasks were significant at the P = 0.05 level, whereas the correlation coefficients for the VAS for the neck and both the left and right trapezii during the stress task approached significance (P = 0.05 and P = 0.1, respectively). These results suggest that decreases in pain observed at long term follow up after myofeedback training might occur as a result of an increased ability to relax but not as a result of decreased muscle activation level. However, the largest correlation found was 0.6. This means that the maximal explained variance (R (2)) is low (36%), and that there are also other processes than the changes in muscle activation that contribute to changes in perceived pain. PMID- 16365786 TI - Genetics, cell cycle and cell expansion in organogenesis in plants. PMID- 16365787 TI - Bimodal distribution of flowering time in a natural hybrid population of daylily (Hemerocallis fulva) and nightlily (Hemerocallis citrina). AB - Time of flower anthesis in a day is thought to evolve in response to the time of pollinator activities. We studied blooming and withering time in natural populations of daylily (Hemerocallis fulva), nightlily (Hemerocallis citrina) and their hybrids, and also in an artificially obtained array of the F1 hybrids. Blooming time of H. fulva varied from 4:30 to 7:30 and H. citrina varied from 16:30 to 20:30. In a natural hybrid population, blooming time and withering time showed discontinuous bimodal distribution in spite that morphological traits of flowers showed continuous unimodal variation. Most F1 hybrids showed diurnal flowering. These findings indicate that only a few genes have strong phenotypic effect on the determination of flowering time in Hemerocallis, and suggest that the evolution from a H. fulva-like ancestor to H. citrina was not a continuous process by accumulation of minute mutations. PMID- 16365788 TI - Recurrent mutation of the KIF21A gene in Japanese patients with congenital fibrosis of the extraocular muscles. AB - PURPOSE: To report recurrent mutation of the KIF21A gene in three Japanese families in which some members have congenital fibrosis of the extraocular muscles type 1 (CFEOM1), and to describe the clinical characteristics of the families. METHODS: Standard ocular examinations were performed on 18 normal and affected members of three unrelated families. To detect mutations, we determined the DNA sequence of exons 8, 20, and 21 and the splice sites of the KIF21A gene. RESULTS: All affected members had a heterozygous mutation of the KIF21A gene in exon 21 (R954W). Clinically, each patient had congenital bilateral ptosis, an infraducted primary position of each eye, and the inability to raise either eye above midline. CONCLUSIONS: The KIF21A gene mutation R954W was detected in the patients with CFEOM1 screened in this study, all of whom were Japanese, reflecting similar reports from Europe, America, the Middle East, and Japan. We suggest that mutations of the KIF21A gene contribute to the development of CFEOM1 regardless of ethnicity. We also found that the delimitation of the KIF21A gene mutation site enabled us to efficiently detect the KIF21A gene mutation despite the large number of KIF21A gene exons. PMID- 16365789 TI - Properties of corneas reconstructed with cultured human corneal endothelial cells and human corneal stroma. AB - PURPOSE: To examine the properties of corneas tissue-engineered with cultured human corneal endothelial cells (HCEC) and human corneal stroma. METHODS: Primary HCEC cultures were established from endothelial cell layer explants and propagated on culture dishes coated with bovine corneal endothelial extracellular matrix. A cell suspension of HCEC at the fifth passage was transferred onto human corneal stroma deprived of endothelial cells, and the corneas were gently centrifuged to enhance cell attachment. The cell density of the tissue-engineered corneas was examined after staining with alizarin red and trypan blue. The tissue engineered corneas were histologically examined by light and electron microscopy. The pump function of the tissue-engineered corneas was measured using an Ussing chamber. RESULTS: The mean endothelial cell density of four tissue-engineered corneas was 2380 +/- 264 cells/mm(2) (mean +/- SD). HCEC on the tissue-engineered corneas had a morphology similar to HCEC in vivo. The pump function parameters of the tissue-engineered corneas were 55%-75% of those of normal corneas. CONCLUSIONS: HCEC on the tissue-engineered corneas have morphology and cellular density similar to HCEC in vivo, whereas the pump function of the tissue engineered corneas was lower than in normal corneas. PMID- 16365791 TI - How spatial orientation of Japanese text affects fixation points in patients with bilateral macular atrophy. AB - PURPOSE: To ascertain the retinal area used by patients with bilateral macular atrophy when reading Japanese text of different character sizes written horizontally or vertically. In addition, to determine fixation points as part of the first of a series of studies designed ultimately to enhance the quality of life of these patients through the improvement of reading acuity. METHODS: Seventeen patients (34 eyes) with bilateral macular atrophy were tested to determine the retinal area employed for reading (R fixation point). Sentences were arranged either horizontally or vertically and projected onto the retina using a scanning laser ophthalmoscope. We also determined the fixation point using microperimetry (M fixation point). The positional relationships between these two fixation points and the scotoma were examined. RESULTS: The R and M fixation points were the same in 20 of the 34 eyes. Multiple R fixation points were found in 11 eyes. The R fixation point was frequently positioned above the lesion when reading horizontally (nine eyes), while it was often positioned in the area nasal to (eight eyes) or temporal to (six eyes) the lesion when reading vertically. CONCLUSIONS: Fixation points changed frequently in these patients with bilateral macular atrophy depending on the spatial orientation of the text. These data should be used in the future to help patients learn how to use the preferred retinal locus to improve their reading skills and enhance their quality of life. PMID- 16365790 TI - Study of effects of antiglaucoma eye drops on N-methyl-D-aspartate-induced retinal damage. AB - PURPOSE: To study the effects of antiglaucoma eye drops on N-methyl-D-aspartate (NMDA)-induced retinal damage. METHODS: Several antiglaucoma eye drops, beta blockers, alpha/beta-blockers, an alpha1-blocker, an alpha2-agonist, and a prostaglandin derivative, were topically administrated to NMDA-treated rat eyes daily for 2 weeks, and the retinal thickness, the number of retrograde-labeled retinal ganglion cells (RGCs), and the results of a cDNA microarray analysis were studied. RESULTS: Intravitreal administration of NMDA caused a significant decrease in the thickness of the retinal layers and induced upregulation of glial fibrillary acidic protein (GFAP). Topical administration of beta-blockers (timolol, betaxolol, and carteolol) and a prostaglandin derivative (latanoprost) showed almost no significant effects on retinal thickness, the number of RGCs, or expression of GFAP. In contrast, the alpha/beta-blockers (nipradilol and levobunolol), the alpha1-blocker (bunazosin HCl), and the alpha2-agonist (brimonidine) showed preservation effects on retinal thickness and the number of RGCs, and marked suppression of NMDA-induced upregulation of GFAP. Among 1101 genes related to cellular regulatory mechanisms, the expression of two genes, both for insulin-like growth factors, (IGF-1) and ErbB3, was altered upon administration of the alpha/beta-blockers, the alpha1-blocker, and the alpha2 agonist. CONCLUSION: Our present study suggests that modulations of the alpha adrenergic receptor, alpha1-blocking and alpha2-stimulation, by antiglaucoma eye drops may cause beneficial effects on NMDA-induced retinal damage in the rat. PMID- 16365792 TI - Topographic characteristics of the optic nerve head measured with scanning laser tomography in normal Japanese subjects. AB - PURPOSE: Few studies have been performed regarding the topographic information obtained with the Heidelberg retina tomograph (HRT) in normal Japanese. In this study, we examined the factors influencing optic disc parameters and hemisphere symmetry obtained with the HRT in normal Japanese subjects. METHODS: Mean values and the standard deviation range for the main HRT parameters were evaluated in 223 eyes of 223 normal Japanese subjects. The influence of age, sex, and disc size on HRT topographic data was analyzed. The superior and inferior hemisphere topographic parameters were compared. RESULTS: Disc area showed a significant difference by sex (P = 0.0493). Rim volume (r = -0.208, P = 0.019), height variation contour (r = -0.275, P = 0.001), and mean retinal nerve fiber layer (RNFL) thickness (r = -0.366, P = 0.001) declined with age. All parameters except height variation contour and mean RNFL thickness showed a clinically significant correlation with disc size (-0.159 < r < 0.719, P < 0.01). Cup parameters in the superior hemisphere were significantly greater than those in the inferior hemisphere. In contrast, rim parameters in the superior hemisphere were significantly smaller than those in the inferior hemisphere. CONCLUSIONS: Some factors, namely, sex, age, and disc size, affected the optic disc parameters in the HRT measurements. Possible parameter asymmetry between the two hemispheres should be considered in normal eyes. PMID- 16365793 TI - Posterior aspect of the orbital septum is reinforced by ligaments. AB - PURPOSE: To investigate the ligaments reinforcing the posterior aspect of the orbital septum. METHODS: Sixteen upper eyelids of eight cadavers of Asians were dissected. Ten were subjected to gross dissections to investigate the ligaments attached to the posterior aspect of the orbital septum and to investigate the relationships with the associated ligamentous structures, and six were used for histological sections to elucidate the ligament anchoring sites in the septum. RESULTS: The ligaments were attached to the posterior aspect of the orbital septum in both upper and lower eyelids in all cases. Some septa in the upper eyelids were supported by the lower-positioned transverse ligament in the lateral area. In all cases, part of the Lockwood ligament was attached to the posterior aspect of the orbital septum in the lower eyelids. Histologically, the ligaments were anchored to the posterior aspect of the orbital septum. CONCLUSIONS: The ligaments were attached to the posterior aspect of the orbital septum. These ligaments, in cooperation with the associated ligaments, are thought to complement the fragility of the orbital septum. PMID- 16365794 TI - Relationship between the s-wave amplitude of the multifocal electroretinogram and the retinal nerve fiber layer thickness in glaucomatous eyes. AB - PURPOSE: To determine whether a significant correlation exists between the amplitude of the s wave of the multifocal electroretinogram (mfERG) and the retinal nerve fiber layer thickness (RNFLT) in glaucomatous eyes. METHODS: Twenty three eyes of 23 patients with glaucoma were studied. In all eyes, the severity of the defects in the upper visual field differed significantly from the severity of those in the lower half. Patients having visual field halves with mean deviation (MD) values lower than -5 dB were placed in group A, and patients having visual field halves with MD values greater than -5 dB were placed in group B. The mfERGs were elicited by 37 stimulus elements alternating from white to black in a pseudorandom binary m-sequence at a frequency of 9.4 Hz. The mfERGs in the upper and lower visual field halves were summed to yield upper and lower averaged waves. The GDx variable corneal compensator and optical coherence tomography were used to measure the RNFLT. RESULTS: The retinal nerve fiber layer was significantly thinner in group A than in group B. There was a significant correlation between the RNFLT and the MD values of visual field defects. The s wave amplitude was significantly smaller in group A than in group B. The s-wave amplitude also correlated significantly with the MD and the RNFLT. CONCLUSION: The significant correlations between the s-wave amplitude and the MD, and between the s-wave amplitude and the RNFLT, indicate that the s-wave receives significant contributions from the retinal ganglion cells and their axons. PMID- 16365795 TI - Application of thermo-setting gel as artificial vitreous. AB - PURPOSE: In this study, we produced a rabbit model and investigated the safety of intravitreous injection of a thermo-setting gel (TG) to determine whether TG can be used as artificial vitreous. METHODS: Ten male Japanese white rabbits were used. After performing vitrectomy in a unilateral eye, we injected 1 ml of WTG 127 into the vitreous cavity. The contralateral control eye was not given ophthalmic solution or surgery. Each eye was examined and intraocular pressure (IOP) and the electroretinogram (ERG) were evaluated. On day 28, all eyes were enucleated and examined. RESULTS: No abnormal findings and no elevation of IOP were observed. On the ERG, no significant difference in the latency and amplitude of either the a wave or b wave was observed. Histopathological examination of the retinal tissue showed no abnormalities. In the presence of a retinal tear, under the detached retina a drift of TG through the tear was observed in a few animals. CONCLUSIONS: In a rabbit model, the safety of using an intravitreous injection of thermo-setting gel as artificial vitreous was confirmed by ophthalmoscopic, electrophysiological, and histological studies for a relatively short observation period. However, TG injection cannot be expected to provide a tamponade effect. PMID- 16365796 TI - Static ocular counterroll: video-based analysis after minimizing the false torsion factors. AB - PURPOSE: To determine the validity and usefulness of a newly developed measurement method of static ocular counterrolling (s-OCR) that eliminates false torsion factors and to test the Jampel hypothesis that s-OCR does not exist. METHODS: A lightweight measurement device, consisting of a video camera, a coaxial light source, and a laser pointer projecting a fixation target on the wall, was fixed to a subject's head by means of a mouthpiece. In 11 healthy adults (mean age: 30 +/- 15 years), digital images of the right eye were captured while the subject kept his head tilted at a randomly selected angle ranging from 0 degrees to 50 degrees . By a frame-by-frame analysis of movements of the corneal light reflex and the iris patterns, OCR was evaluated. RESULTS: Torsional eye movement in the opposite direction to head tilt was found in all subjects. The amount of torsion continuously increased until the head-tilt angle reached 40 degrees. The average (+/- SD) amplitude of a fitted sine curve was 7.6 +/- 3.2 degrees (range: 4.3 degrees-10.3 degrees), and the individual amplitude was significantly larger than the test-retest repeatability of the measurement (+/ 1.7 degrees). CONCLUSIONS: The measurement method used in this study provided good test-retest repeatability and ease of application. The characteristics of torsional eye movements that we observed after minimizing the false-torsion factors agree with previous reports supporting the existence of s-OCR. PMID- 16365797 TI - Orbital pseudotumor as an initial manifestation of multicentric Castleman's disease. AB - BACKGROUND: To report on a patient with multicentric Castleman's disease (MCD) who showed an orbital pseudotumor as an initial manifestation. CASE: A Japanese male patient, 65 years old, initially exhibited swelling of the left eyelid. Computed tomography and magnetic resonance imaging showed an infiltrating orbital mass around the left eye. OBSERVATIONS: The patient was treated several times with short courses of oral corticosteroids without significant response. Eight years after the first examination, he was referred to our neuroophthalmology clinic owing to persistent eyelid swelling. He exhibited an accelerated erythrosedimentation rate, polyclonal hypergammaglobulinemia, and multiple cervical and thoracic mediastinal lymphadenopathies. Histopathological examination revealed mixed-type MCD from a cervical lymph node and chronic inflammatory cell infiltration from eyelid swelling. Four months after his first visit, the patient's condition was complicated by fever, general fatigue, and hypoxemia. A cell marker study of the swollen lymph node showed monoclonality, indicating the development of non-Hodgkin's lymphoma. CONCLUSIONS: The clinical course in this case suggests that an inflammatory reaction in the orbital tissue was initially present, before MCD, and that non-Hodgkin's lymphoma subsequently developed. PMID- 16365798 TI - Trans-Tenon's retrobulbar injection of triamcinolone acetonide for diffuse diabetic macular edema. AB - PURPOSE: To determine whether a trans-Tenon's retrobulbar injection of triamcinolone acetonide (TA) is a safe and effective treatment for diffuse diabetic macular edema. METHODS: Thirty-nine eyes of 30 diabetic patients with persistent macular edema were treated with 20 mg of TA injection. Central macular thickness (CMT) determined by optical coherence tomography (OCT) and visual acuity were evaluated before the injection and at 1, 2, 3, and 6 months, and up to 1 year in some eyes, after the injection. RESULTS: The CMT decreased significantly from 478 +/- 129 microm (mean +/- SD) before injection to 316 +/- 102 microm at 1 month, 307 +/- 104 microm at 2 months, and 275 +/- 89 microm at 3 months after a single injection of TA. A 20% reduction of CMT from the initial value was maintained by a single injection of TA in 27 of 39 eyes (69.2%) at 3 months, in 14 of 22 eyes (63.6%) at 6 months, and in 5 of 7 eyes at 12 months. A recurrence of macular edema was observed in 10% of the eyes at 3 months, and in 22.7% at 6 months. The 17 eyes in which vitrectomy had been carried out had a more significant improvement in CMT than the eyes without vitrectomy. CONCLUSION: A 20-mg trans-Tenon's retrobulbar TA injection is a safe and effective treatment for diabetic macular edema. PMID- 16365799 TI - Ring-shaped subretinal fibrinous exudate in central serous chorioretinopathy. AB - PURPOSE: To clarify the pathologic changes of the ring-shaped, grayish white subretinal fibrinous exudate in central serous chorioretinopathy (CSC), we examined patients with fluorescein angiography (FA) and optical coherence tomography (OCT). METHODS: Using OCT, we prospectively examined 13 consecutive patients (12 men, one woman) (13 eyes) with acute CSC. The fundi were scanned by OCT through the ring-shaped, grayish white lesion. RESULTS: FA showed focal dye leakage at the center of the ring-shaped, grayish white subretinal exudates. In the area of the exudates, OCT showed moderately or highly reflective masses bridging the neurosensory retina and the retinal pigment epithelium in all eyes. Because the area of the pellucid core of the ring-shaped exudate showed an optically clear space beneath the neurosensory retina, the subretinal reflective masses were separated on cross-sectional images in all 13 eyes. Pigment epithelium detachment was observed in the area of the grayish white lesion by OCT and FA in 7 of the 13 eyes. CONCLUSIONS: In eyes with a ring-shaped, grayish white exudate in CSC, OCT and FA findings indicate that the pellucid core of the exudate is the route of the exudative fluid from the choroid into the subretinal space. A surrounding grayish white lesion seems to be fibrinous exudate that has accumulated in the subretinal space. PMID- 16365800 TI - A nontraumatic macular hole in a 10-year-old girl. AB - BACKGROUND: Full-thickness macular holes usually develop in the elderly population. To the best of our knowledge, there has been no written report of a nontraumatic macular hole in a pediatric patient. CASE: A 10-year-old girl noticed decreased central vision in her left eye without any history of trauma. OBSERVATIONS: Fundus examination of the left eye revealed a full-thickness macular hole and a thin fibrous membrane on the superior peripapillary retina. She underwent standard macular hole surgery with stripping of the membrane, resulting in closure of the hole. CONCLUSIONS: A full-thickness macular hole may develop in pediatric patients. Although the etiology of the macular hole in the present patient is unclear, tangential traction induced by contraction of the peripapillary membrane, presumed to be an incomplete regression of the Bergmeister papilla, might have been responsible for the formation of the macular hole. PMID- 16365801 TI - Comparative study on the combined effects of bunazosin and nipradilol or timolol on intraocular pressure in normotensive rabbits. AB - PURPOSE: To compare the ocular hypotensive effect of nipradilol and timolol in combination with bunazosin in rabbits. METHODS: The intraocular pressure (IOP) in normal rabbits was measured using an applanation pneumatonograph. Nipradilol, timolol, and bunazosin were instilled, individually or in combination, into the inferior conjunctival sac. RESULTS: Nipradilol (0.25%), timolol (0.5%), and bunazosin (0.01%) individually lowered IOP. The IOP-lowering effects of both nipradilol and timolol were significantly enhanced by the combined application of bunazosin (0.01%). In the presence of 5% timolol or 0.1% bunazosin, IOP was further lowered by the addition of nipradilol. The IOP-lowering effect of nipradilol was partly inhibited by pretreatment with c-PTIO (10 mM), a nitric oxide (NO)-trapping agent. CONCLUSIONS: The present study demonstrated that the IOP-lowering effects of nipradilol are due to beta- and alpha1-blocking and NO donating actions, and bunazosin has an additive effect on the IOP-lowering effect of nipradilol or timolol. PMID- 16365802 TI - Massive macular hard exudates associated with branch retinal vein occlusion. AB - PURPOSE: To report four cases of branch retinal vein occlusion (BRVO) with the complication of serous retinal detachment (SRD). METHODS: We retrospectively studied four eyes of four patients with macular edema and macular hard exudates associated with midperipheral BRVO. Visual acuity, ophthalmoscopy, fluorescein angiography, and optical coherence tomography findings were evaluated. Three of the four eyes underwent laser photocoagulation in the BRVO area 1 month after the initial visit. RESULTS: Macular edema consisted of SRD without cystoid macular edema in all eyes. Late-phase fluorescein angiography showed extensive dye leakage in the BRVO area. When SRD was resolved 4 months after the initial examination, hard exudates had increased in the macular area. Although macular hard exudates decreased 1 year after the initial examination, visual acuity remained under 20/20 because of macular atrophy. CONCLUSIONS: SRD is one type of macular edema observed in BRVO. In macula-spared midperipheral BRVO, the SRD originates from a vascular leaking area, and there is a high risk that massive macular hard exudates will develop, which may affect visual recovery. PMID- 16365803 TI - Relation of posterior staphyloma in highly myopic eyes with macular hole and retinal detachment. AB - PURPOSE: To investigate whether the type of posterior staphyloma (PS) affects the development of macular hole and retinal detachment (MHRD) in high myopia. METHODS: Included in this retrospective study were 28 highly myopic eyes with MHRD (MHRD group) and 47 highly myopic eyes without MHRD (control group). The presence or absence of PS and the type of PS according to the classification of Curtin, the postoperative visual acuity, and the retinal reattachment rate were investigated. RESULTS: The percentage of eyes with PS was significantly higher in the MHRD group than in the control group (P < 0.001). The rate of type II PS was significantly higher in the MHRD group (P = 0.01). There were no significant differences among the types of PS regarding the retinal reattachment rates and visual outcome. CONCLUSIONS: Eyes with type II PS have a higher risk for the development of MHRD. However, the type of PS did not seem to affect the anatomic and visual outcome. PMID- 16365804 TI - Charles Bonnet syndrome associated with a first attack of multiple sclerosis. AB - BACKGROUND: We treated a rare case of Charles Bonnet syndrome (CBS) manifested during temporary blindness in both eyes caused by optic neuritis associated with a first attack of multiple sclerosis (MS). CASE: A 66-year-old Japanese woman became completely blind for 3 months due to optic neuritis after a first attack of MS. During the blind period, she experienced vivid visual hallucinations for about 2 weeks. OBSERVATIONS: The patient had no psychiatric disorders or cognitive impairments; therefore, the visual hallucinations during the period of blindness were indicative of CBS. Unexpectedly, the hallucinations disappeared without treatment following her recovery of vision. CONCLUSIONS: Although rare, visual impairment during a first attack of MS can be associated with visual hallucinations indicative of CBS. The hallucinations can disappear spontaneously with the recovery of vision without treatment. PMID- 16365805 TI - Permanent loss of vision following cosmetic rhinoplastic surgery. PMID- 16365806 TI - Herpes zoster panuveitis progression despite acyclovir treatment in a patient following bone marrow transplantation. PMID- 16365808 TI - A young Japanese man with relentless placoid chorioretinitis. PMID- 16365807 TI - Bacterial endophthalmitis after triamcinolone acetonide-assisted pars plana vitrectomy. PMID- 16365809 TI - Intermuscular transverse ligament goes under the orbital part of the lacrimal gland and attaches to the lateral orbital wall. PMID- 16365810 TI - A case of Mooren's ulcer associated with a pterygium. PMID- 16365811 TI - Iridociliary melanocytoma with suspected pulmonary metastasis. PMID- 16365812 TI - Central serous retinal detachment detected by optical coherence tomography in Leber's idiopathic stellate neuroretinitis. PMID- 16365813 TI - Sclerokeratitis in Takayasu's arteritis: a case report. PMID- 16365815 TI - Laparoscopic hepatectomy: indications and outcomes. AB - We outline the indications, evaluate the degree of invasiveness, and analyze the outcomes of laparoscopic hepatectomy, mainly in the treatment of hepatocellular carcinoma (HCC). The important considerations in determining indications for laparoscopic hepatectomy include tumor size, type, and location. Nodular tumors smaller than 4 cm or pedunculated tumors smaller than 6 cm are suitable candidates. Concerning location, tumors in the lower segment or the left lateral segment are suitable. Regarding operative method, laparoscopic hepatectomy involving either partial hepatectomy or left lateral segmentectomy is a feasible, less invasive procedure. Operative time in our recent laparoscopic hepatectomy patients has decreased, with less bleeding. Furthermore, laparoscopic hepatectomy is less invasive than conventional hepatectomy on evaluation by the Estimation of Physiolic Ability and Surgical Stress (E-PASS) scoring system. Patients recovered more quickly after laparoscopic hepatectomy, which allowed shorter hospitalization. Both the 5-year survival rate for HCC and the survival rate without recurrence were nearly identical to those of open conventional hepatectomy, although further analysis will be necessary to reach definitive conclusions. In conclusion, laparoscopic hepatectomy avoids the disadvantages of standard hepatectomy in properly selected patients and is beneficial for patient quality of life, because it is a minimally invasive procedure when indications are strictly followed. PMID- 16365814 TI - Analysis of risk factors for massive intraoperative bleeding during laparoscopic splenectomy. AB - BACKGROUND/PURPOSE: Laparoscopic splenectomy is occasionally converted to open surgery due to massive intraoperative bleeding. The aim of this study was to identify the risk factors for massive bleeding during laparoscopic splenectomy. METHODS: Fifty-three patients underwent laparoscopic splenectomy. The indications were hematologic disease in 25 patients, liver cirrhosis in 17 patients, and other conditions in 11 patients. Univariate analysis was conducted with Fisher's exact test, and multivariate analysis was conducted with a stepwise logistic regression model. RESULTS: None of the patients required open surgery. Blood loss of more than 800 ml was defined as massive intraoperative bleeding. Univariate analysis showed significant risk factors for massive bleeding to be liver cirrhosis, portal hypertension, splenomegaly, Child class, and preoperative platelet count. Independent risk factors in the multivariate analysis were portal hypertension and Child class. CONCLUSIONS: Careful attention to intraoperative bleeding during laparoscopic splenectomy is necessary for patients with portal hypertension and/or deteriorated liver function. PMID- 16365816 TI - Technical aspects in the laparoscopic management of complicated common bile duct stones. AB - The management of common bile duct (CBD) stones traditionally required open laparotomy and bile duct exploration. With the advent of endoscopic and laparoscopic technology in the latter half of last century, endoscopic retrograde cholangiopancreatography (ERCP) and laparoscopic cholecystectomy (LC) has become the mainstream treatment for CBD stones and gallstones in most medical centers around the world. However, in certain situations, ERCP cannot be feasible because of difficult cannulation and extraction. ERCP can also be associated with potential serious complications, in particular for complicated stones requiring repeated sessions and additional maneuvers. Since our first laparoscopic exploration of the CBD (LECBD) in 1995, we now adopt the routine practice of the laparoscopic approach in dealing with endoscopically irretrievable CBD stones. The aim of this article is to describe the technical details of this approach and to review the results from our series. PMID- 16365817 TI - Laparoscopic pancreatic surgery. AB - In the past, in the pancreas, a minimally invasive technique was only used for diagnostic laparoscopy in evaluating periampullary malignancy. Recent advances in operative techniques and instrumentation have empowered surgeons to perform virtually all procedures in the pancreas, including the Whipple procedure. Some of these procedures represent the most sophisticated application of minimally invasive surgery, and their outcomes are reportedly better than those of conventional open approaches. In addition to the evaluation of resectability in periampullary malignancy, palliative procedures, including biliary bypasses and gastrojejunostomy, can be performed laparoscopically. Although it is reportedly feasible to perform a Whipple procedure laparescopically, no benefit of the laparoscopic approach over the conventional open approach has been documented. Laparoscopic distal pancreatectomy, with or without preserving the spleen, is technically easier than the Whipple procedure, and is more widely accepted. Indications for laparoscopic distal pancreatectomy include cystic neoplasms and islet-cell tumors located in the pancreatic body or tail. Complications of acute and chronic pancreatitis may be treated with the use of surgical laparoscopy. When infected necrotizing pancreatitis is identified, surgical intervention for drainage and debridement is required. According to the type and location of infected necrotizing pancreatitis, three laparoscopic operative approaches have been reported: infracolic debridement, retroperitoneal debridement, and laparoscopic transgastric pancreatic necrosectomy. When internal drainage is indicated for a pseudocyst, a minimally invasive technique is a promising option. Laparoscopic pseudocyst gastrostomy, cyst jejunostomy, or cyst duodenostomy can be performed, depending on the size and location of the pseudocyst. Especially when a pseudocyst is located in close contact with the posterior wall of the stomach, it is best drained by a pseudocyst gastrostomy, which can also be done with the use of an intragastric operative technique. PMID- 16365818 TI - Controversies in staging of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the most common primary cancer of the liver. As the natural history of HCC is better delineated, treatment strategies are in constant evolution. Classic staging systems based on histopathology following resection are often inadequate because the majority of patients present with advanced disease. The dissatisfaction with anatomic staging systems has resulted in the emergence of several new clinical staging systems, which attempt to integrate tumor biology and the underlying function of the nondiseased liver. Tragically, the byproduct of multiple staging strategies is confusion for physicians, patients, and clinical researchers; this undermines the principles on which staging systems are created. To address this issue, a consensus conference was organized in 2002 to identify the best staging strategy for HCC. The purpose of this article is to review the current clinical and pathologic staging strategies and to highlight the recommendations from the consensus conference. PMID- 16365819 TI - Hepatolithiasis and intrahepatic cholangiocarcinoma: carcinogenesis based on molecular mechanisms. AB - Hepatolithiasis is more frequently seen in East Asian countries than in Western countries, and it is well known to represent a high-risk state for intrahepatic cholangiocarcinoma. Intrahepatic cholangiocarcinoma is an aggressive tumor that shows a dismal outcome even after resection. Cancer results from multistep carcinogenesis; however, the precise molecular mechanisms involved in the genetic alterations in cancer remain unknown. The accumulation of alterations in cancer related genes leads to disruptions in cell-cycle regulation and also to continuous cell proliferation. The present review provides an overview of cancer related genes in intrahepatic cholangiocarcinogenesis arising in hepatolithiasis. Further study of molecular mechanisms in hepatolithiasis-related intrahepatic cholangiocarcinoma, and the delineation of the influence of the genes involved should lead to our understanding of cholangiocarcinogenesis. PMID- 16365820 TI - Continuous vertical hemimattress suture for biliary-enteric anastomosis. AB - The continuous hemivertical mattress suture technique for biliary-enteric anastomosis has not been well reported in the literature. We used the technique with a double-armed monofilament absorbable suture (Glycomer 631) for 32 anastomoses in 31 patients. There was one anastomotic leakage (3.1%). The mean follow-up period was 683 days, during which time no patient developed anastomotic stenosis or cholangitis. The technique provided satisfactory results with lower cost than one-layer interrupted sutures. PMID- 16365821 TI - Peritoneal "pillowcase" for the displaced spleen post-distal pancreatectomy. AB - BACKGROUND PURPOSE: Spleen-preserving distal pancreatectomy, although rarely performed, has become an option for treatment of distal pancreatic neoplasms. However, the spleen, once mobilised, is often free to tort. A reliable method of fixing the spleen would be a useful adjunct to this procedure. METHODS: We describe the use of a new technique to fix the spleen using a peritoneal "pillowcase" to envelop the spleen, thus preventing its torsion. RESULTS: This patient has done well, with no problems encountered intraoperatively or postoperatively, using this new technique of spleen fixation. CONCLUSIONS: The use of a peritoneal "pillowcase" to envelope the spleen following distal pancreatectomy has proven a useful adjunct to this somewhat difficult surgical procedure. It may be applied in other cases where spleen fixation is necessary. PMID- 16365822 TI - Laparoscopic gastric bypass for gastric outlet obstruction is associated with smoother, faster recovery and shorter hospital stay compared with open surgery. AB - BACKGROUND/PURPOSE: Laparoscopic gastric bypass for relief of gastric outlet obstruction (GOO) is feasible and safe. However, comparative data to confirm the benefits of the laparoscopic approach remain scarce. METHODS: Between 1998 and 2003, 26 patients underwent 15 laparoscopic (surgeon A) and 12 open (surgeon B) gastrojejunostomies (GJs) for GOO. The indications for surgery included malignant (n = 17) and benign (n = 10) diseases. RESULTS: There were no conversions to open surgery in the laparoscopic group, and no operative mortality occurred in either group. The groups were comparable for age, sex, American Society of Anesthesiology (ASA) score, frequencies of previous abdominal surgery and of malignant or benign disease, and type of GJ fashioned. There were no differences between the laparoscopic and open groups with regard to the operating time (median, 90 vs 111 min; P = 0.113), and patients receiving intraoperative blood transfusion. However, laparoscopic surgery was associated with significantly shorter durations of postoperative intravenous hydration (60 vs 234 h; P = 0.001), opiate analgesia (49 vs 128 h; P = 0.025), and hospital stay (3 vs 15 days; P = 0.005). Operative morbidity occurred more frequently following open surgery (33% vs 13%; P = 0.219). CONCLUSIONS: Laparoscopic GJ for the relief of GOO is associated with a smoother and more rapid postoperative recovery and shorter hospital stay compared with open surgery. In experienced hands, the laparoscopic approach to GJ should become the new gold standard. PMID- 16365823 TI - Serosal invasion in TNM staging of mass-forming intrahepatic cholangiocarcinoma. AB - BACKGROUND/PURPOSE: The Liver Cancer Study Group of Japan established a tumor nodule-metastasis (TNM) staging system for mass-forming intrahepatic cholangiocarcinoma, with T determined by tumor number and size and vascular or serosal invasion. Serosal invasion is not considered in the designation established by the International Union Against Cancer. METHODS: Sixty-three patients who underwent hepatic resection for mass-forming intrahepatic cholangiocarcinoma were investigated retrospectively, with the investigation including univariate and multivariate analyses of potential prognostic factors. RESULTS: By log-rank test, tumor size more than 3.0 cm, vascular invasion, lymph node metastasis, intrahepatic metastasis, and involved resection margin, but not serosal invasion, were associated significantly with poor prognosis. Even in patients with serosal invasion, the postoperative outcome was much better in those without than in those with vascular invasion. Multivariate analysis identified vascular invasion, lymph node metastasis, and an involved resection margin as independent prognostic factors. When serosal invasion was excluded from tumor staging, the 5-year survival rates became more clearly stratified: 100% in those with stage I disease, 62% in those with stage II, 25% in those with stage III, and 7% for patients with stage IV. CONCLUSIONS: Serosal invasion showed no survival impact after hepatic resection for mass-forming intrahepatic cholangiocarcinoma. When serosal invasion was omitted from the TNM staging proposed by the Liver Cancer Study Group of Japan, stratification of postoperative survival between stages was more effective. PMID- 16365825 TI - Laparoscopic hepatic resection for completely exophytic hepatocellular carcinoma on cirrhosis. AB - Laparoscopic surgery is a relatively new option for the treatment of hepatocellular carcinoma (HCC) on cirrhosis. To date, there have been only a few reports of this option for this pathology in the literature, probably because of the intra operative difficulties related to the treatment of this pathology (even at laparotomy) and because of the problems related to the minimally invasive approach (technical difficulties, complicated management of the bleeding, lack of dedicate tools, and fear of gas embolism). In this article we report four patients from our whole series (23 laparoscopic liver resections for HCC) who underwent a laparoscopic resection for completely exophytic HCC on cirrhosis, located in segment IV in two patients, and in segment III and segment V respectively, in the other two. The mean operative time was 116 min (range, 90 150 min). The Pringle maneuver was never performed. No blood transfusions were needed. No postoperative complications occurred, neither ascites, nor jaundice, nor encephalopathy. Postoperative liver function returned to the preoperative level within 3 days. Food intake started on postoperative day 2. The patients were discharged on postoperative days 5 (one patient), 6 (two patients), and 7 (one patient) after uncomplicated courses. In our opinion, limited laparoscopic liver resections could be considered, at present, to be the best option for the treatment of extremely rare protruding HCC on cirrhosis. We believe that a minimally invasive approach can minimize the postoperative morbidity rate, which is still too high in this group of patients. Our experience confirmed that nonanatomical limited resections or anatomical left lateral segmentectomies for HCC on cirrhosis are feasible and safe in the hands of surgeons trained in both open liver surgery and advanced laparoscopic surgery. PMID- 16365824 TI - Primary liver cancer with bidirectional differentiation into hepatocytes and biliary epithelium. AB - A 70-year old Japanese man with hepatitis B infection developed a 4.8-cm liver tumor in the right lobe. Computed tomographic findings suggested hepatocellular carcinoma or combined hepatocellular and cholangiocarcinoma. After right hepatectomy, the cut surface of the resected specimen showed a whitish tumor with indistinct borders. Microscopically, the tumor consisted of pleomorphic, eosinophilic cells forming thickened trabeculae, suggesting hepatocellular carcinoma, and also tumor cells forming glandular structures, suggesting adenocarcinoma. Cells comprising both the trabeculae and the glands were immunoreactive for cytokeratin (CK) 19. In some gland-forming cells, varying immunoreactivity for CK7 and carcinoembryonic antigen (CEA) was observed. In the cytoplasm of cells in the thickened trabeculae, hepatocyte paraffin (HepPar) 1 and alpha-fetoprotein (AFP) were focally reactive. Some of these tumor cells were also reactive for CK19. Accordingly, we concluded that this tumor was a combined hepatocellular and cholangiocarcinoma whose cells had undergone divergent differentiation into hepatocytes and biliary epithelium. Because the tumor expressed markers of both phenotypes, it may have originated from cells intermediate between hepatocytes and biliary epithelium. A few reported cases have shown similar histologic features. PMID- 16365826 TI - Mucinous cystic neoplasm of the pancreas during pregnancy: the importance of proper management. AB - We describe a case of huge mucinous cystic tumor of the pancreas in a 26-year-old woman during pregnancy. Ultrasonography demonstrated a well-delimited cystic mass in the left upper abdominal quadrant, suggestive of benignity. Magnetic resonance imaging showed a large cystic mass resembling a mucinous cystic tumor of the pancreas. After this assessment the patient underwent surgical exploration and a huge cystic tumor of the pancreas was discovered. The tumor was enucleated and distal pancreatectomy was performed. The resected margin of the specimen was free of tumor. In this case report we discuss the management of mucinous pancreatic tumors during pregnancy and we briefly review the previously reported cases of mucinous pancreatic tumors in pregnant patients. We conclude that surgical resection of these tumors should be strongly considered in pregnancy. Removal of the tumor appears to be a safe procedure without harmful effects to the fetus. PMID- 16365827 TI - Improvement of surgical margin with a coupled saline-radio-frequency device for multiple colorectal liver metastases. AB - Complete resection of colorectal liver metastases (LM) has been the only curative treatment. However, when LM are multiple and bilobar, only a few patients are candidates for curative surgery. We report on a 53-year-old woman with synchronous multiple and bilobar LM from sigmoidal cancer who became resectable after a multimodal strategy including preoperative systemic chemotherapy and two step surgery. The spectacular decrease in tumor size after systemic chemotherapy led us to perform two-step surgery, including right portal-vein ligation and left liver metastasectomies, with a coupled saline-radiofrequency device, in order to improve the surgical margin. An extended right hepatectomy was performed later to remove the remaining right liver lesions. The patient was discharged after 28 days without major complication and was recurrence-free 14 months later. We conclude that improving the surgical margin with a coupled saline-radiofrequency device is feasible and effective, avoiding small remnant liver even after multiple tumorectomies. The multimodal strategy, including preoperative chemotherapy, two-step surgery, and tumorectomies, using a coupled saline radiofrequency device, could increase the number of patients with diffuse bilobar liver metastases who can benefit from liver resection. PMID- 16365828 TI - Mesenchymal hamartoma of the liver in adulthood: immunohistochemical profiles, clinical and histopathological features in two patients. AB - Mesenchymal hamartoma is an uncommon cystic mass of the liver which occurs primarily in children. There are a few reports of its occurrence in adulthood. Here, we present two cases in female patients, 54 and 51 years old. Radiological examinations in both patients showed multiple cystic lesions in the liver. Surgically, total cystectomy was performed in the first patient, while an unroofing procedure was done in the second patient (due to misdiagnosis of the lesion as a simple cyst of the liver). On microscopic examinations of the lesion in each patient, a multilocular cyst was observed, lined by flattened epithelium and surrounded by a mesenchymal component composed of mature connective tissue, arterial and venous vascular structures, peripheral nerve bundles, and ductal structures. An immunohistochemical panel consisting of desmin, smooth-muscle actin, S-100, vimentin, CD34, carcinoembryonic antigen, pancytokeratin, cytokeratin 7, cytokeratin 8, cytokeratin 17, cytokeratin 18, cytokeratin 19, and cytokeratin 20 was applied to paraffin sections. Immunoreactivity for cytokeratin 7 and cytokeratin 19 was observed in cystic epithelium and ductal structures. Focal and patchy desmin immunoreactivity was observed in connective tissue. S-100 was positive only in peripheral nerve bundles. In conclusion, mesenchymal hamartoma of the liver in adulthood is a localized tumoral abnormality that precedes birth, and which has delayed clinical presentation. These lesions seems to be related to a maturation process. During this period of maturation, immature edematous stroma rich in mucopolysaccharides may convert to mature paucicellular hyalinized connective tissue. This maturation process may be also related to loss of premalignant potential of these tumors. PMID- 16365829 TI - Emotional support levels can predict physical functioning and health related quality of life among elderly Taiwanese with hip fractures. AB - INTRODUCTION: There has been increasing interest in the role emotional support plays on recovery and the perceptions of health. However, the vast majority of studies have been based on data from Western nations. Little is known about hip fractured elders in Asian countries. This study has examined the influences of emotional support on functional recovery, and health-related quality of life (HRQoL) among elderly patients in Taiwan. METHODS: Data were gathered from 126 hip fractured elders, in a teaching hospital in northern Taiwan. Eleven items from the Medical Outcome Study (MOS) Social Support Survey were administered, in order to measure emotional support at 1 month after discharge. The Chinese Barthel Index (CBI), Lawton and Brody's instrumental activities of daily living (IADL) scale, and MOS Short Form-36 Taiwan version were administered at the end of months 1, 3, and 6 after discharge. RESULTS: After controlling for covariates, greater levels of emotional support predicted a better recovery of the activities of daily living (ADL) (beta=0.23, P=0.04) and IADL (beta=0.03, P=0.03), and better HRQoL in seven of eight dimensions (P ranged from <0.0001 to 0.04), except social function, emotional role , and physical role during the first 6 months after hospital discharge. CONCLUSIONS: Emotional support can have a significant impact on the recovery of self- care ability, and different aspects of HRQoL during the first 6 months after discharge for hip fractured elders in Taiwan. Clinical implications were made, on the basis of these findings. PMID- 16365830 TI - Relationship of circulating MMP-2, MMP-1, and TIMP-1 levels with bone biochemical markers and bone mineral density in postmenopausal Chinese women. AB - INTRODUCTION: Osteoblast-derived matrix metalloproteinase (MMP)-2, MMP-1 and tissue inhibitor of metalloproteinase (TIMP)-1 have been shown to play a role in bone metabolism by degrading the bone matrix. METHODS: The present study was performed to investigate the relationships between serum MMP-2, MMP-1, or TIMP-1 levels and bone mineral density (BMD), as well as bone biochemical markers, in 297 Chinese postmenopausal women aged 42-80 years. RESULTS: We found a significant negative weak correlation between MMP-2 and BMD at various skeletal regions. After adjustment for age and BMI, the correlation with BMD at the femoral neck and total hip disappeared. Multiple linear stepwise regression analysis showed that MMP-2 was not a determinant factor for BMD. The significant positive correlations between MMP-2 and bone cross-linked N-telopeptides of type I collagen (NTX), alkaline phosphatase (BAP), and osteocalcin (OC) and were found, and remained significant after adjustment for age and BMI. Moreover, serum MMP-2 concentrations were significantly higher in postmenopausal women with osteoporosis than in age-matched normal controls. There were no significant correlations between MMP-1, TIMP-1 and BMD. There were no significant relationships between MMP-1 and BAP, OC, and NTX. The associations between TIMP-1 and BAP and OC were not specific and constant. CONCLUSIONS: In conclusion, our results suggest that circulating MMP-2 and markers of bone turnover are correlated, and serum MMP-2 levels may rise with increase in bone turnover. PMID- 16365831 TI - A possible smoking susceptibility locus on chromosome 11p12: evidence from sex limitation linkage analyses in a sample of Australian twin families. AB - Many twin studies have identified sex differences in the influence of genetic and environmental factors on smoking behaviors. We explore the evidence for sex differences for smoking initiation and cigarette consumption in a sample of Australian twin families, and extend these models to incorporate sex differences in linkage analyses for these traits. We further examine the impact of including or excluding non-smokers in genetic analyses of tobacco consumption. Accounting for sex differences improved linkage results in some instances. We identified one region suggestive of linkage on chromosome 11p12. This locus, as well as another region identified on chromosome 6p12, replicates regions identified in previous studies. PMID- 16365832 TI - New approaches to the genetic analysis of neuroticism and anxiety. AB - The completion of the human genome project and the complementary genome projects for other species has broadened the scope for novel bioinformatic approaches to quantitative trait locus (QTL) identification. A key issue for quantitative trait nucleotide (QTN) identification is progressing from a large QTL peak, spanning perhaps 50 cM and many hundreds of genes, to a gene or nucleotide variant which is responsible for that QTL effect. The complementary use of mouse models to dissect large syntenic loci in humans is a powerful method for reducing QTL intervals to the order of 1 Mb. This paper presents an overview of the approaches used in our laboratory to ultra-fine map QTLs for anxiety-related traits, and to identify quantitative trait genes (QTG). As new genetic techniques and statistical approaches arise, we are getting closer to identifying those long sought after QTNs. PMID- 16365834 TI - Heteroditopic Rebek's imide directs the reactivity of homoditopic olefins within desolvated quaternary assemblies in the solid state. PMID- 16365833 TI - Regression-based association analysis with clustered haplotypes through use of genotypes. AB - Haplotype-based association analysis has been recognized as a tool with high resolution and potentially great power for identifying modest etiological effects of genes. However, in practice, its efficacy has not been as successfully reproduced as expected in theory. One primary cause is that such analysis tends to require a large number of parameters to capture the abundant haplotype varieties, and many of those are expended on rare haplotypes for which studies would have insufficient power to detect association even if it existed. To concentrate statistical power on more-relevant inferences, in this study, we developed a regression-based approach using clustered haplotypes to assess haplotype-phenotype association. Specifically, we generalized the probabilistic clustering methods of Tzeng to the generalized linear model (GLM) framework established by Schaid et al. The proposed method uses unphased genotypes and incorporates both phase uncertainty and clustering uncertainty. Its GLM framework allows adjustment of covariates and can model qualitative and quantitative traits. It can also evaluate the overall haplotype association or the individual haplotype effects. We applied the proposed approach to study the association between hypertriglyceridemia and the apolipoprotein A5 gene. Through simulation studies, we assessed the performance of the proposed approach and demonstrate its validity and power in testing for haplotype-trait association. PMID- 16365835 TI - Sphingolactones: selective and irreversible inhibitors of neutral sphingomyelinase. PMID- 16365836 TI - Derivatives of octaethynylphenazine and hexaethynylquinoxaline. PMID- 16365837 TI - Fluoromorphic substrates for fatty acid metabolism: highly sensitive probes for mammalian medium-chain acyl-CoA dehydrogenase. PMID- 16365838 TI - Unusual lone pairs in tellurium and their relevance for superconductivity. PMID- 16365839 TI - Biocatalytic synthesis of poly(L-lactide) by native and recombinant forms of the silicatein enzymes. PMID- 16365840 TI - Thermosensitive core-shell particles as carriers for ag nanoparticles: modulating the catalytic activity by a phase transition in networks. PMID- 16365841 TI - Nanohybrids composed of quantum dots and cytochrome P450 as photocatalysts. PMID- 16365842 TI - Label-free, real-time glycosyltransferase assay based on a fluorescent artificial chemosensor. PMID- 16365843 TI - DNA-templated self-assembly of two-dimensional and periodical gold nanoparticle arrays. PMID- 16365844 TI - Formation of cyclic sulfinates and sulfinamides through homolytic substitution at the sulfur atom. PMID- 16365845 TI - Synthesis of 2,4,6-trimethylphenylcalcium iodide and degradation in THF solution. PMID- 16365846 TI - Superlinear scaling in master-slave quantum chemical calculations using in-core storage of two-electron integrals. AB - We describe the implementation of a parallel, in-core, integral-direct Hartree Fock and density functional theory code for the efficient calculation of Hartree Fock wave functions and density functional theory. The algorithm is based on a parallel master-slave algorithm, and the two-electron integrals calculated by a slave are stored in available local memory. To ensure the greatest computational savings, the master node keeps track of all integral batches stored on the different slaves. The code can reuse undifferentiated two-electron integrals both in the wave function optimization and in the evaluation of second-, third-, and fourth-order molecular properties. Superlinear scaling is achieved in a series of test examples, with speedups of up to 55 achieved for calculations run on medium sized molecules on 16 processors with respect to the time used on a single processor. PMID- 16365847 TI - Characterization and redox properties of cytochrome c552 from Thermus thermophilus adsorbed on different self-assembled thiol monolayers, used to model the chemical environment of the redox partner. AB - The structure of cytochrome c552 (Cyt-c552) from Thermus thermophilus shows many differences to other c-type cytochromes. The rich lysine domain close to the heme does not exist in this cytochrome, allowing us to postulate that the interaction with its redox partner must be different to the cytochrome c/cytochrome c oxidase interaction. We report a study of Cyt-c552 adsorbed on self-assembled monolayers (SAMs) of functionalized alkanethiols used to mimic the chemical properties of its redox partner (ba3-oxydase). Hydrophilic (-COOH), polar (-OH), hydrophobic ( CH3), and mixed (-OH/-CH3) SAMs grafted on roughened silver electrodes were characterized by X-ray photoelectron spectroscopy. Surface enhanced resonance Raman spectroscopy (SERRS) was employed to determine the structure and the redox properties (E degrees and number of transferred electron) of the heme of Cyt-c552 adsorbed on roughened silver electrodes coated by the different SAMs. The surface that most closely models the environment of the ba3-oxidase is a mixed SAM formed by 50% polar [Ag-(CH2)5-CH2OH] and 50% hydrophobic [Ag-(CH2)5-CH3] alkanethiols. Only the native form B1(6cLS) of Cyt-c552 is detected by SERRS when the protein is adsorbed on such a surface that promotes a protein orientation favorable for the electron transfer (number of transferred electron = 1). We shall discuss the differences and similarities of the electron-transfer mechanism of Cyt-c552 compared to cyt-c. PMID- 16365848 TI - Investigating oligonucleotide hybridization at subnanomolar level by surface plasmon resonance biosensor method. AB - We have optimized surface plasmon resonance (SPR) biosensor technology for a rapid, direct, and low-consumption label-free multianalyte screening of synthetic oligonucleotides (ONs) with modified internucleotide linkages potentially applicable in antisense therapy. Monitoring of the ONs hybridization is based on the formation of complex between the natural oligonucleotide probe immobilized on the sensor surface and the ON in solution in contact with the sensor surface. An immobilization chemistry utilizing the streptavidin-biotin interaction was employed to obtain desired ligand density and high hybridization efficiency. It was demonstrated that the sensor is capable of detecting complementary 23-mer ONs in concentrations as low as 0.1 nM with high specificity and reproducibility. PMID- 16365849 TI - E230Q mutation of the catalytic subunit of cAMP-dependent protein kinase affects local structure and the binding of peptide inhibitor. AB - The active site of the mammalian cAMP-dependent protein kinase catalytic subunit (C-subunit) has a cluster of nonconserved acidic residues-Glu127, Glu170, Glu203, Glu230, and Asp241-that are crucial for substrate recognition and binding. Studies have shown that the Glu230 to Gln mutant (E230Q) of the enzyme has physical properties similar to the wild-type enzyme and has decreased affinity for a short peptide substrate, Kemptide. However, recent experiments intended to crystallize ternary complex of the E230Q mutant with MgATP and protein kinase inhibitor (PKI) could only obtain crystals of the apo-enzyme of E230Q mutant. To deduce the possible mechanism that prevented ternary complex formation, we used the relaxed-complex method (Lin, J.-H., et al. J Am Chem Soc 2002, 24, 5632-5633) to study PKI binding to the E230Q mutant C-subunit. In the E230Q mutant, we observed local structural changes of the peptide binding site that correlated closely to the reduced PKI affinity. The structural changes occurred in the F-to G helix loop and appeared to hinder PKI binding. Reduced electrostatic potential repulsion among Asp241 from the helix loop section and the other acidic residues in the peptide binding site appear to be responsible for the structural change. PMID- 16365850 TI - Theoretical studies on electron delocalization in diaminoguanidine. AB - Ab initio MO and density functional methods have been employed to understand the electronic structure and second-order delocalizations in diaminoguanidine. The potential energy surfaces of the three different positional isomers of diaminoguanidine (DAG1, DAG2, and DAG3) have been studied in detail, which suggest greater stability for DAG1 over the other isomers. Natural Population Analysis (NPA) has been employed to understand the primary and the secondary electron delocalizations in diaminoguanidine. The absolute proton affinity of diaminoguanidine is greater than both guanidine as well as aminoguanidine due to the extra stability from the intramolecular interactions in protonated diaminoguanidine. The increase in the electron delocalization upon protonation in diaminoguanidine is similar to that of guanidine and aminoguanidine. PMID- 16365851 TI - Cutaneous melanoma in postmenopausal women after nonmelanoma skin carcinoma: the Women's Health Initiative Observational Study. AB - BACKGROUND: An elevated risk for cutaneous melanoma has been reported in individuals with nonmelanoma skin carcinoma (NMSC), but to the authors' knowledge, this association has not been prospectively studied in a large, multigeographic population of postmenopausal women. METHODS: The association between NMSC and the incidence of cutaneous melanoma was assessed in the Women's Health Initiative Observational Study involving 67,030 non-Hispanic white postmenopausal women ages 50-79 years and who were free of prior other cancers at baseline. Cancer history, demographics, and previous and current risk exposures were determined by questionnaires at baseline and follow-up. Participants' reports of incident cutaneous melanoma collected annually were confirmed by physician review of medical records. Cox proportional hazards analyses were used to assess the relation of prior NMSC with incident cutaneous melanoma. RESULTS: In age-adjusted analysis, women with a history of NMSC but no other malignancy (n = 5552) were found to be 2.41 times more likely to develop cutaneous melanoma over a mean 6.5 years compared with women who had no history of NMSC (95% confidence interval [95% CI], 1.82-3.20). In a multivariate analysis, women with a history of NMSC and no other cancer history at baseline were 1.70 times more likely to develop cutaneous melanoma compared with women without NMSC (95% CI, 1.18-2.44). CONCLUSION: The results of the current study provide evidence and further defines the magnitude of increased risk for cutaneous melanoma in postmenopausal non-Hispanic white women with a history of NMSC. PMID- 16365852 TI - Adjuvant therapy in pediatric patients with completely resected renal cell carcinoma. PMID- 16365853 TI - A syndrome of irreversible leukoencephalopathy following pediatric allogeneic bone marrow transplantation. AB - BACKGROUND: Despite decreases in overall mortality following bone marrow transplantation (BMT), a number of complications such as neurotoxicity have been described and often associated with immunosuppressive agents. The syndrome of reversible posterior leukoencephalopathy has been described in patients receiving cyclosporin and FK-506. We report here a subset of children who developed a syndrome of previously undescribed irreversible leukoencephalopathy following allogeneic BMT. PATIENTS AND METHODS: Between 1996 and 2002, 138 pediatric patients received an allogeneic BMT at Lucile Salter Packard Children's Hospital at Stanford. Six cases of irreversible leukoencephalopathy were observed. Cases were defined as children who exhibited progressive and continued, severe neurologic deterioration lasting greater than 2 weeks and consistent with non localizing, central nervous system abnormalities. Medical records and magnetic resonance images (MRIs) were reviewed. RESULTS: Median age of the affected patients at BMT was 7.8 years. All six received cyclosporine, and [corrected] one had elevated drug levels. Encephalopathy occurred at a median of 53 days (range 14-77) following BMT. Symptoms at onset of leukoenceophalopathy included confusion and altered mental status, sluggish pupillary responses, abnormal movements, and seizures. Two patients died during their neurologic decline. Four patients remain alive with persistent encephalopathy. MRI showed abnormalities in all patients including periventricular or subcortical white matter involvement in all, and basal ganglia lesions in three. CONCLUSIONS: We report a syndrome of irreversible neurologic deficits and cerebral white matter abnormalities following allogeneic BMT, yet not associated with elevated cyclosporin levels. A precise mechanism for this syndrome is lacking and warrants further consideration. PMID- 16365854 TI - Population demography of Northern muriquis (Brachyteles hypoxanthus) at the Estacao Biologica de Caratinga/Reserva particular do Patrimonio Natural-Feliciano Miguel Abdala, Minas Gerais, Brazil. AB - The 957-ha forest at the Estacao Biologica de Caratinga/Reserva Particular do Patrimonio Natural-Feliciano Miguel Abdala, in Minas Gerais, Brazil, supports one of the largest known populations of the critically endangered northern muriqui (Brachyteles hypoxanthus). We combine long-term data on one group that has been monitored since 1982 with new data obtained on the other three groups since 2002 and 2003 to describe the demographic structure of this population, evaluate its potential for future growth, and predict how dispersal and competitive regimes may change in response to current demographic conditions. As of January 2005, the 226 individuals in the population were divided into four mixed-sex groups with 37 77 members, and an all-male unit whose eight males maintained transient associations with two of the mixed-sex groups. Although 51.77% of the population was female, the sex ratio among adults and subadults was female-biased (0.75), while that among immatures was male-biased (1.47). Consistent with expectations from mean interbirth intervals, 64.18% of adult females gave birth in 2003 and 2004. However, by January 2005, only 52.31% of adult females were still carrying infants <24 months of age due to unusually high infant mortality. First-year survivorship among the 25 infants born in the population in 2003 was only 76%, considerably lower than previously documented in the longest-studied group. High female fecundity is indicative of a healthy population, but the current male bias in births will result in a decline in the population growth rate within two decades, and may increase levels of male reproductive competition and alter dispersal patterns. PMID- 16365855 TI - Fractures in late medieval skeletal populations from Serbia. AB - Bone fractures were analyzed from skeletal remains of 861 adult individuals from six cemeteries dating to the Late Medieval period in Serbia. Results of the study were compared to other cemetery populations (635 individual skeletons) of the same date and region in an attempt to understand fracture patterns. The association of types of fractures and their prevalence with sex, age at death, cemetery site, and information deriving from historical sources are discussed. Results showed that the long bone fracture frequency was 0.7%, and the majority of the fractures were the result of direct force. This rate is similar to some studies of contemporary British skeletal samples. However, it is much lower than for some other Old World sites. Cranial vault fractures had a rate of 6.7%, and of the facial skeleton, 1.3%; the frontal bone was the most affected of bones of the cranial vault. Injuries were more common on the upper extremities (0.8%) compared to the lower (0.6%). However, the fibula was the most fractured bone (2.8%), followed by the ulna (2.4%). This pattern is similar to three of six Late Medieval urban sites in Britain. These findings suggest that this rural community was exposed to a low risk of trauma, probably related mostly to accidents sustained during farming, and rarely to interpersonal violence. PMID- 16365856 TI - Do human parents face a quantity-quality tradeoff?: evidence from a Shuar community. AB - A number of evolutionary theories of human life history assume a quantity-quality tradeoff for offspring production: parents with fewer offspring can have higher biological fitness than those with more. Direct evidence for such a tradeoff, however, is mixed. We tested this assumption in a community of Ecuadorian Shuar hunter-horticulturalists, using child anthropometry as a proxy for fitness. We measured the impact of household consumer/producer (CP) ratio on height, weight, skinfold thicknesses, and arm and calf circumferences of 85 children and young adults. To control for possible "phenotypic" correlates that might mask the effect of CP ratio on anthropometry, we also measured household garden productivity, wealth, and social status. Regression models of the age standardized variables indicated a significant negative impact of CP ratio on child growth and nutrition. The age-standardized height and weight of children in households with the largest CP ratio (10) were 1.38 and 1.44 standard deviations, respectively, below those of children in households with the smallest CP ratio (2). Surprisingly, garden productivity, wealth, and status had little to no effect on the fitness proxies. There was, however, an interesting and unexpected interaction between status and sex: for females, but not males, higher father status correlated significantly with higher values on the proxies. PMID- 16365857 TI - Cortical bone remodeling rates in a sample of African American and European American descent groups from the American Midwest: comparisons of age and sex in ribs. AB - This study employs regression analysis to explore population and sex differences in the pattern of age-associated bone loss, as reflected by histomorphometric variables that are measures of intracortical and endocortical bone remodeling. A comparison of an African American sample from the Washington Park Cemetery in St. Louis, Missouri, and a European American rib sample composed of cadavers, autopsies, and forensic cases from Missouri reveals the existence of complex age associated patterns for differences in measures of intracortical remodeling and cortical area. Females from the two samples express similar bone dimensions and dynamics. The African American females appear to lose more bone than their male counterparts, but this difference is absent in the European American sample. When age-associated patterns are considered, it is in the younger cohorts that African Americans exhibit greater relative cortical area than European Americans, but this is reversed in the older ages, when the latter group manifests greater bone mass. The European American males consistently differ in the slopes and intercepts for the variables compared to the other groups, and differences are highly significant with African American females, with the former group maintaining bone mass while the latter exhibit a more rapid bone loss. Achieving larger relative cortical area due to smaller endosteal area, coupled with better bone quality due to lower intracortical porosity early in life, may be a mechanism by which African Americans, especially females, maintain adequate bone mass in older ages, which buffers them from bone loss and related fragility fractures despite higher rates of intracortical remodeling and endosteal expansion later in life. These results suggest that both genetic and environmental factors are responsible for the differences in bone remodeling and bone mass observed between these samples. PMID- 16365858 TI - Conservation biology of Malagasy strepsirhines: a phylogenetic approach. AB - The phylogenetic diversity of extant lemurs represents one of the most important but least studied aspects of the conservation biology of primates. The phylogenetic diversity of a species is inversely proportional to the relative number and closeness of its phylogenetic relatives. Phylogenetic diversity can then be used to determine conservation priorities for specific biogeographic regions. Although Malagasy strepsirhines represent the highest phylogenetic diversity among primates at the global level, there are few phylogenetic data on species-specific and regional conservation plans for lemurs in Madagascar. Therefore, in this paper the following questions are addressed for extant lemurs: 1) how does the measure of taxonomic uniqueness used by Mittermeier et al. (1992 Lemurs of Madagascar; Gland, Switzerland: IUCN) equate with an index of phylogenetic diversity, 2) what are the regional conservation priorities based on analyses of phylogenetic diversity in extant lemurs, and 3) what conservation recommendations can be made based on analyses of phylogenetic diversity in lemurs? Taxonomic endemicity standardized weight (TESW) indices of phylogenetic diversity were used to determine the evolutionary component of biodiversity and to prioritize regions for conserving lemur taxa. TESW refers to the standardization of phylogenetic diversity indices for widespread taxa and endemicity of species. The phylogenetic data came from recent genetic studies of Malagasy strepsirhines at the species level. Lemur species were assigned as being either present or absent in six biogeographic regions. TESW indices were combined with data on lemur complementarity and protected areas to assign conservation priorities at the regional level. Although there were no overall differences between taxonomic ranks and phylogenetic rankings, there were significant differences for the top-ranked taxa. The phylogenetic component of lemur diversity is greatest for Daubentonia madagascariensis, Allocebus trichotis, Lepilemur septentrionalis, Indri indri, and Mirza coquereli. Regional conservation priorities are highest for lemurs that range into northeast humid forests and western dry forests. Expansion of existing protected areas in these regions may provide the most rapid method for preserving lemurs. In the long term, new protected areas must be created because there are lemur species that: 1) are not found in existing protected areas, 2) exist only in one or two protected areas, and 3) are still being discovered outside the current network of protected areas. Data on the population dynamics and feeding ecology of phylogenetically important species are needed to ensure that protected areas adequately conserve lemur populations in Madagascar. PMID- 16365859 TI - Health of infants in an Imperial Roman skeletal sample: perspective from dental microstructure. AB - This study examines general health in the first year of life of a population of 127 subadults from the Imperial Roman necropolis of Isola Sacra (2nd-3rd century ACE). Health status was determined by analyzing 274 deciduous teeth from these children for Wilson bands (also known as accentuated striae), microscopic defects caused by a disruption to normal enamel development arising from some generalized external stressor. While macroscopic enamel defects, or hypoplasias, have long been used as proxies of general population health, we believe that this is the first population-wide study of microscopic defects in deciduous teeth. We used microstructural markers of enamel to attach very precise chronologies to Wilson band formation that allowed us to calculate maximum prevalence (MAP) and smoothed maximum prevalence (SMAP) distributions to portray what we believe to be a realistic risk profile for a past population of children. There appear to be two periods of high prevalence, the first beginning around age 2 months and continuing through month 5, and the second higher period beginning around month 6 and continuing through month 9. These results are discussed in light of historical records of Roman childhood rearing practices. PMID- 16365860 TI - On the trail of pulmonary tuberculosis based on rib lesions: results from the Human Identified Skeletal Collection from the Museu Bocage (Lisbon, Portugal). AB - In the last 20 years, studies on human identified skeletal collections have revealed a significant relationship between new bone formation on the visceral surface of ribs and pulmonary tuberculosis (TB). To improve methods of differential diagnosis of respiratory diseases in archaeological skeletons, an investigation was conducted on 197 individuals from the Human Identified Skeletal Collection of the Museu Bocage (Lisbon, Portugal). This sample included 109 males and 88 females who lived during the 19th-20th centuries, with ages at death ranging from 13-88 years. The skeletons were grouped according to cause of death: 1) pulmonary TB (N = 84); 2) pulmonary non-TB diseases (N = 49); and 3) a control group (N = 64) composed of individuals randomly selected among the extrapulmonary non-TB causes of death. The ribs, sterna, scapulae, and clavicles were macroscopically observed. New bone formation on the visceral surface of ribs was recorded in 90.5% (76/84) of individuals who died from pulmonary TB, in 36.7% (18/49) with a pulmonary non-TB disease as cause of death, and in 25.0% (16/64) of the control group. These differences were statistically significant (P < 0.001). Furthermore, in individuals with pulmonary TB, the bony lesions presented mainly as lamellar bone on the vertebral end of the upper and middle thoracic rib cage. Proliferative alterations also occurred on one sternum and in nine clavicles and eight scapulae. This work strongly supports the results of similar studies performed on other documented collections, suggesting that new bone formation on ribs, although not pathognomonic, is a useful criterion for the differential diagnosis of pulmonary TB. PMID- 16365861 TI - Effect of mobility on femur midshaft external shape and robusticity. AB - This study investigates differences in femur midshaft shape, robusticity, and sexual dimorphism derived from external measurements between a broad range of prehistoric and historic North American populations with different subsistence strategies and inferred levels of mobility. The sample was divided into six groups to test whether observed femur midshaft variables follow the patterns predicted based on archaeologically and historically determined subsistence and mobility data. The results suggest significant variation in femur midshaft shape and robusticity in all populations, and that inferred mobility levels do not correspond consistently with femur midshaft structure in either males or females. Results do, however, support the prediction that sexual dimorphism is generally greater in more mobile populations. PMID- 16365862 TI - Childhood acute lymphoblastic leukemia in the age of genomics. AB - The recent sequencing of the human genome and technical breakthroughs now make it possible to simultaneously determine mRNA expression levels of almost all of the identified genes in the human genome. DNA "chip" or microarray technology holds great promise for the development of more refined, biologically-based classification systems for childhood ALL, as well as the identification of new targets for novel therapy. To date gene expression profiles have been described that correlate with subtypes of ALL defined by morphology, immunophenotype, cytogenetic alterations, and response to therapy. Mechanistic insights into treatment failure have come from the definition of mRNA signatures that predict in vitro chemoresistance, as well as differences between blasts at relapse and new diagnosis. New bioinformatics tools optimize data mining, but validation of findings is essential since "over-fitting" the data is a common danger. In the future, genomic analysis will be complemented by evaluation of the cancer proteome. PMID- 16365863 TI - Correlation between phenotypic heterogeneity and gene mutational characteristics in familial hemophagocytic lymphohistiocytosis (FHL). AB - BACKGROUND: Classification of familial hemophagocytic lymphohistiocytosis (FHL) into FHL2, FHL3, and other subtypes based on genetic abnormalities has recently become possible. We studied the phenotypic differences among these subtypes in Japan. METHODS: Forty patients clinically diagnosed with FHL were analyzed. Perforin abnormality was screened by flow cytometric analysis and/or DNA sequencing in these patients, and those without perforin abnormalities were further examined for the presence of mutations in the Munc13-4 gene by DNA sequencing. The correlation between clinical features and genetic subtypes was investigated. RESULTS: Of the 40 HLH patients, 11 showed perforin gene mutations (classified as FHL2) and ten had Munc13-4 gene mutations (FHL3), but neither mutation was noted in 19 patients (non-FHL2/3). Although the majority of the patients developed the disease before the age of 1 year, the onset in three FHL2 patients with missense mutations was late (7, 11, and 12 years). Incidence of deficient natural killer cell activity was higher in FHL2 patients (9/9 FHL2, 4/9 FHL3, and 6/17 non-FHL2/3; P = 0.005). The serum levels of ferritin and soluble interleukin-2 receptor were significantly higher in FHL2 patients with nonsense perforin mutations compared to other subgroups (P < or = 0.05). Epstein-Barr virus infection was involved in 8 of the 40 HLH patients: one FHL2, one FHL3, and six non-FHL2/3. CONCLUSIONS: Although clinical features of FHL3 appear to be homogeneous, the heterogeneous clinical features of FHL2 depend upon the nature of perforin gene mutations. Characterization of the non-FHL2/3 group with regard to FHL1 or other novel gene mutations remains to be conducted. PMID- 16365864 TI - A case series of pediatric primary central nervous system lymphoma: favorable outcome without cranial irradiation. AB - PURPOSE: To describe the outcome of childhood primary central nervous system lymphoma (PCNSL) treated with chemotherapy alone or with chemotherapy plus cranial radiotherapy (CRT). METHODS: Retrospective chart review of children with PCNSL at six tertiary care pediatric centers. RESULTS: Eight immunocompetent and four immunocompromised children were included. Ten children received chemotherapy alone without CRT, with most receiving high-dose methotrexate and high-dose cytarabine. Five year event-free survival (EFS) in this group was 70.0 +/- 14.5%. Two children received chemotherapy plus CRT; one relapsed and died while the other is alive in remission. Three children died, including two from relapsed disease. The other child with human immunodeficiency virus infection died of an opportunistic infection while in remission following chemotherapy alone. CONCLUSIONS: Most children with PCNSL can achieve long-term remissions with chemotherapy alone without CRT. Multi-center prospective studies are needed to confirm or refute these results in a larger number of patients. PMID- 16365865 TI - May-Thurner syndrome (iliac vein compression) and thrombosis in adolescents. AB - May-Thurner syndrome refers to anatomic compression of the left iliofemoral vein by the overriding right iliac artery. We report three adolescents who presented to our pediatric hospital with iliac vein thrombosis and were diagnosed with May Thurner syndrome. Each received catheter-directed thrombolysis followed by balloon angioplasty to restore flow. Two patients had endovascular stents placed. The procedures were well tolerated, without major complications. Additional thrombophilic risk factors were identified in each patient. Though uncommon, pediatric hematologists should consider May-Thurner syndrome in adolescents who present with a left lower extremity thrombosis. Aggressive therapy may be warranted due to the risk of post-thrombotic syndrome. PMID- 16365866 TI - Effect of storage upon material properties of lyophilized porcine extracellular matrix derived from the urinary bladder. AB - Xenogeneic extracellular matrices (ECMs) have been developed as off-the-shelf biologic scaffolds that have been effectively used in preclinical and clinical applications for tissue reconstruction. Such materials must be suitable for terminal sterilization and capable of storage for extended periods of time without significant changes in material properties and bioactivity. Material properties of interest for ECM scaffolds include hydrostatic permeability index (PI), uniaxial maximum load and elongation, maximum tangential stiffness (MTS), suture retention strength (SRS), and ball-burst strength (BBS). The present study evaluated these material properties for lyophilized forms of an ECM scaffold derived from the porcine urinary bladder, termed urinary bladder matrix (UBM), that was terminally sterilized by e-beam irradiation at 22 kGy and stored at room temperature (RT; 20-24 degrees C) or refrigerated temperature (REFT; 4-8 degrees C) for up to 12 months. UBM devices showed no change in SRS, BBS, and hydrostatic PI after the evaluation period. Lyophilized devices stored at RT showed an increase in maximum load and MTS while devices stored at REFT showed an increase in maximum elongation after 1 year of storage (p < 0.05). These results indicate that structural changes in the UBM device may slowly occur as a function of prolonged storage and storage temperature. PMID- 16365867 TI - Structure, magnetizability, and nuclear magnetic shielding tensors of bis heteropentalenes. IV. Dihydrophospholophosphole isomers. AB - The geometry of the heteropentalenes formed by two phosphole units has been determined at the DFT level. The magnetic susceptibility and the nuclear magnetic shielding at the nuclei of these systems have also been calculated using gauge including atomic orbitals and a large Gaussian basis set to achieve near Hartree Fock estimates. A comparative study of the various isomers, of their flattened analogs, and of the parent phosphole molecule, shows that the [3,4-c] isomer is the most aromatic system in the set considered, assuming diatropicity and degree of planarity as indicators, even if it is the less stable in terms of total molecular energy. Plots of magnetic field-induced current densities confirm diatropicity of P-containing bis-heteropentalenenes, showing, however, significant differences from the analogous systems with distinct heteroatoms. The maps give evidence of spiral flow nearby C-C bonds, compatible with prevalent distortive behavior of pi electrons exalted by pyramidalization at P, and competing against the sigma electron compression, which would favor planar structure. PMID- 16365868 TI - Gaseous reaction mechanism of C2F radical with water. AB - The kinetic properties of the carbon-fluorine radicals are little understood except those of CFn (n =1-3). In this article, a detailed mechanistic study was reported on the gas-phase reaction between the simplest pi-bonded C2F radical and water as the first attempt to understand the chemical reactivity of the C2F radical. Various reaction channels are considered. The most kinetically competitive channel is the quasi-direct hydrogen-abstraction route forming P5 HCCF + OH. At the CCSD(T)/6-311+G(2d,2p)//B3LYP/6-311G(d,p)+ZPVE, CCSD(T)/6 311+G(3df,2p)//QCISD/6-311G(d,p)+ZPVE and Gaussian-3//B3LYP/6-31G(d) levels, the overall H-abstraction barriers (4.5, 4.7, and 4.2 kcal/mol) for the C2F + H2O reaction are comparable to the corresponding values (5.5, 3.7, and 5.7 kcal/mol) for the analogous C2H + H2O reaction. This suggests that C2F is a reactive radical like the extensively studied C2H, in contrast to the situation of the CF and CF2 radicals that have much lower reactivity than the corresponding hydrocarbon species. Thus, the C2F radical is expected to play an important role in the combustion processes of the carbon-fluorine chemistry. Furthermore, addition of a second H2O can catalyze the reaction with the H-abstraction barrier significantly reduced to a marginally zero value (0.5 kcal/mol). This is also indicative of the potential relevance of the title reactions in the low temperature atmospheric chemistry. PMID- 16365869 TI - Parallel coupled perturbed CASSCF equations and analytic CASSCF second derivatives. AB - A parallel algorithm for solving the coupled-perturbed MCSCF (CPMCSCF) equations and analytic nuclear second derivatives of CASSCF wave functions is presented. A parallel scheme for evaluating derivative integrals and their subsequent use in constructing other derivative quantities is described. The task of solving the CPMCSCF equations is approached using a parallelization scheme that partitions the electronic hessian matrix over all processors as opposed to simple partitioning of the 3 N solution vectors among the processors. The scalability of the current algorithm, up to 128 processors, is demonstrated. Using three test cases, results indicate that the parallelization of derivative integral evaluation through a simple scheme is highly effective regardless of the size of the basis set employed in the CASSCF energy calculation. Parallelization of the construction of the MCSCF electronic hessian during solution of the CPMCSCF equations varies quantitatively depending on the nature of the hessian itself, but is highly scalable in all cases. PMID- 16365870 TI - Mutations of perforin and Munc13-4 do not mark HLH by NK defects. PMID- 16365871 TI - Migraine and MTHFR C677T genotype in a population-based sample. AB - OBJECTIVE: Migraine with aura is associated with increased risk of stroke. The MTHFR C677T genotype has been associated with increased risk of migraine in selected clinical samples and with elevated homocysteine, a risk factor for stroke. We assessed the association of the MTHFR C677T variant with migraine and the mediating effect of cardiovascular risk factors and metabolic markers of genotype status. METHODS: We compared adult migraineurs with aura (MA; n = 187), without aura (MO; n = 226), and nonmigraineurs (n = 1,212) from the population based Genetic Epidemiology of Migraine study. RESULTS: Compared with the wild type genotype, the T/T genotype was associated with increased odds of MA (odds ratio [OR], 2.05; 95% confidence interval, 1.2-3.4; p < 0.006), with a trend of increasing numbers of T alleles (OR, 1.40; 95% confidence interval, 1.1-1.8; p < 0.007). ORs were slightly attenuated after adjusting for homocysteine. INTERPRETATION: Risk of MA is associated with MTHFR C674T homozygosity, independent of other cardiovascular risk factors. PMID- 16365872 TI - SEPN1: associated with congenital fiber-type disproportion and insulin resistance. AB - OBJECTIVE: Our first objective was to determine whether SEPN1 gene mutations are a cause of congenital fiber-type disproportion (CFTD), a rare form of congenital myopathy in which relative hypotrophy of type 1 (slow twitch) muscle fibers is the principal abnormality on histology. Second, we investigated an association between SEPN1-related myopathy and insulin resistance. METHODS: We sequenced SEPN1 in five unrelated CFTD patients with scoliosis and respiratory muscle weakness and screened an additional 22 CFTD patients for abnormalities in SEPN1 by Western blotting and restriction digest for the 943G-->A mutation. We performed oral glucose tolerance tests (OGTTs) in eight SEPN1-related myopathy patients. RESULTS: Two sisters with CFTD were homozygous for the 943G-->A SEPN1 mutation and had clinical features typical of previously reported patients with SEPN1-related myopathy. Five of eight SEPN1-related myopathy patients had abnormalities on OGTT suggestive of insulin resistance. INTERPRETATION: SEPN1 is the second genetic cause of CFTD and the first cause of autosomal recessive CFTD to be identified to our knowledge. CFTD is the fourth clinicopathological presentation that can be associated with mutations in SEPN1. Insulin resistance may be a specific, previously unrecognized aspect of SEPN1-related myopathy. PMID- 16365873 TI - Molecular basis of interaction between NG2 proteoglycan and galectin-3. AB - Previous work has demonstrated the ability of the NG2 proteoglycan, a component of microvascular pericytes, to stimulate endothelial cell motility and morphogenesis. This function of NG2 depends on formation of a complex with galectin-3 and alpha3beta1 integrin to stimulate integrin-mediated transmembrane signaling. In addition, the co-expression of galectin-3 and NG2 in A375 melanoma cells suggests that the malignant properties of these cells may be affected by interaction between the two molecules. Here, we extend the theme of co-expression and interaction of NG2 and galectin-3 to human glioma cells. We also establish a molecular basis for the NG2/galectin-3 interaction. The C-terminal carbohydrate recognition domain of galectin-3 is responsible for binding to the NG2 core protein. Within the NG2 extracellular domain, the membrane-proximal D3 segment of the proteoglycan contains the primary binding site for interaction with galectin 3. The interaction between galectin-3 and NG2 is a carbohydrate-dependent one mediated by N-linked rather than O-linked oligosaccharides within the D3 domain of the NG2 core protein. These studies establish a foundation for attempts to reduce the aggressive properties of tumor cells by disrupting the NG2/galectin-3 interaction. PMID- 16365874 TI - Cellular and molecular mechanisms of nicotine's pro-angiogenesis activity and its potential impact on cancer. AB - The present study examined the mechanisms of nicotine's effect on angiogenesis and its impact on tumor growth. Nicotine demonstrated significant (P<0.01) stimulation of the release of endothelial cell growth factor, basic fibroblast growth factor (b-FGF) but not vascular endothelial growth factor (VEGF). In a concentration-dependent manner, nicotine induced endothelial cell tube formation. Additionally, in the chick chorioallantoic membrane (CAM) model of angiogenesis, nicotine effectively induced the generation of new blood vessels (P<0.01), an effect that is mediated via b-FGF. The pro-angiogenesis effect of nicotine in the CAM model was maximally blocked by either anti-integrin alphavbeta3 or inhibitor of mitogen activated protein kinase (MAPK, ERK 1/2). In the CAM tumor implant model, nicotine doubled (P<0.01) the growth rate of breast, colon, and lung cancer. These data indicated that the pro-angiogenesis effect is mediated via b FGF and induced through the nicotinic receptor, alphavbeta3 integrin, and MAPK. PMID- 16365875 TI - Rad9, an evolutionarily conserved gene with multiple functions for preserving genomic integrity. AB - The Rad9 gene is evolutionarily conserved. Analysis of the gene from yeast, mouse and human reveal roles in multiple, fundamental biological processes primarily but not exclusively important for regulating genomic integrity. The encoded mammalian proteins participate in promoting resistance to DNA damage, cell cycle checkpoint control, DNA repair, and apoptosis. Other functions include a role in embryogenesis, the transactivation of multiple target genes, co-repression of androgen-induced transcription activity of the androgen receptor, a 3'-5' exonuclease activity, and the regulation of ribonucleotide synthesis. Analyses of the functions of Rad9, and in particular its role in regulating and coordinating numerous fundamental biological activities, should not only provide information about the molecular mechanisms of several individual cellular processes, but might also lend insight into the more global control and coordination of what at least superficially present as independent pathways. PMID- 16365876 TI - Chromatin remodeling in DNA replication. AB - Chromatin remodeling complexes have evolved to solve a very basic problem for eukaryotic cells accommodation of the genome to fit the dimensions of the nucleus without loss of access to the DNA molecule. In the nucleus, DNA is wrapped around histones to form nucleosomes and other higher order compact chromatin structures. Chromatin remodeling complexes enable highly regulated access to DNA sequences in the context of chromatin, and it is well known that these complexes are involved in regulation of transcription. However, gene expression is not the only process that occurs in the nucleus. DNA has to be replicated, recombined, and repaired. In this regard, it is notable that the recent discoveries have linked ATP dependent remodeling complexes to DNA damage repair. These results have raised challenging questions about the possible versatility of chromatin remodeling complexes in other nuclear activities, particularly in DNA replication, since a number of recent studies have suggested a connection between this essential cellular process and chromatin remodeling. However, the chromatin remodeling events regulating DNA replication have not been extensively investigated. The aim of this prospect is to summarize recent studies that implicate chromatin remodeling in DNA replication and to address potential roles of chromatin remodeling at various stages of eukaryotic DNA replication. PMID- 16365877 TI - Gas-phase ion chemistry in organometallic systems. AB - This review essentially deals with positive ion/molecule reactions occurring in gas-phase organometallic systems, and encompasses a period of time of approximately 7 years, going from 1997 to early 2004. Following the example of the excellent review by Eller & Schwarz (1991; Chem Rev 91:1121-1177), in the first part, results of reaction of naked ions are presented by grouping them according to the neutral substrate, while in the second part, ligated ions are grouped according to the different ligands. Whenever possible, comparison among similar studies is attempted, and general trends of reactivities are evidenced. PMID- 16365878 TI - Microtubule disassembly and inhibition of mitosis by a novel synthetic pharmacophore. AB - Microtubule drugs, which block cell cycle progression through mitosis, have seen widespread use in cancer chemotherapies. Although microtubules are subject to regulation by signal transduction mechanisms, their pharmacological modulation has so far relied on compounds that bind to the tubulin subunit. A new microtubule pharmacophore, diphenyleneiodonium, causing disassembly of the microtubule cytoskeleton is described here. Although this synthetic compound does not affect the assembly state of purified microtubules, it profoundly suppresses microtubule assembly in vivo, causes paclitaxel-stabilized microtubules to cluster around the centrosomes, and selectively disassembles dynamic microtubules. Similar to other microtubule drugs, this new pharmacophore blocks mitotic spindle assembly and mitotic cell division. PMID- 16365879 TI - UVB-induced production of 1,25-dihydroxyvitamin D3 and vitamin D activity in human keratinocytes pretreated with a sterol delta7-reductase inhibitor. AB - The skin fulfills an important role in the vitamin D photo-endocrine system. Epidermis is not only the site of vitamin D3 photoproduction. In addition, epidermal keratinocytes contain the vitamin D receptor (VDR) and possess 25 hydroxylase and 1alpha-hydroxylase activity indicating that all components of the vitamin D system are present. We investigated whether these components cooperate in inducing vitamin D activity upon treatment with physiological UVB doses. Upon irradiation, 24-hydroxylase mRNA was induced in keratinocytes pretreated with a sterol Delta7-reductase inhibitor (BM15766) whereby the 7-dehydrocholesterol content increased by 300-fold. Transfection experiments with a vitamin D response element containing construct confirmed VDR-dependent gene activation. Furthermore, the UVB-dependent induction of 24-hydroxylase was blocked by the cytochrome-P450 inhibitor ketoconazole. The 24-hydroxylase inducing photoproduct was transferable to unirradiated keratinocytes by medium and cellular homogenates of UVB-irradiated, BM15766-pretreated cells and was identified as 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] by high-performance liquid chromatography with tandem mass spectrometric detection. Addition of vitamin D binding protein blunted UVB-induced 24-hydroxylase suggesting the possibility of a paracrine or autocrine role for 1,25(OH)2D3. In conclusion, epidermal keratinocytes can produce vitamin D3, convert it to 1,25(OH)2D3 and respond to it upon UVB irradiation in the absence of exogenous 7-dehydrocholesterol and therefore contain a unique and complete photo-endocrine vitamin D system. PMID- 16365880 TI - Subcomplexes of human ATP synthase mark mitochondrial biosynthesis disorders. AB - OBJECTIVE: METHODS: We describe biochemically and clinically relevant aspects of mitochondrial ATP synthase, the enzyme that supplies most ATP for the cells energy demand. RESULTS: Analyzing human Rho zero cells we could identify three subcomplexes of ATP synthase: F1 catalytic domain, F1 domain with bound natural IF1 inhibitor protein, and F1-c subcomplex, an assembly of F1 domain and a ring of F(O)-subunits c. Large amounts of F1 subcomplexes accumulated also in mitochondria of patients with specific mitochondrial disorders. By quantifying the F1 subcomplexes and other oxidative phosphorylation complexes in parallel, we were able to discriminate three classes of defects in mitochondrial biosynthesis, namely, mitochondrial DNA depletion, mitochondrial transfer RNA (tRNA) mutations, and mutations in the mitochondrial ATP6 gene. INTERPRETATION: The relatively simple electrophoretic assay used here is a straightforward approach to differentiate between various types of genetic alterations affecting the biosynthesis of oxidative phosphorylation complexes and will be useful to guide molecular genetic diagnostics in the field of mitochondrial neuromuscular disorders. PMID- 16365881 TI - Caspase inhibition in apoptotic T cells triggers necrotic cell death depending on the cell type and the proapoptotic stimulus. AB - CD95 (Fas/Apo-1) triggers apoptotic cell death via a caspase-dependent pathway. Inhibition of caspase activation blocks proapoptotic signaling and thus, prevents execution of apoptosis. Besides induction of apoptotic cell death, CD95 has been reported to trigger necrotic cell death in susceptible cells. In this study, we investigated the interplay between apoptotic and necrotic cell death signaling in T cells. Using the agonistic CD95 antibody, 7C11, we found that caspase inhibition mediated by the pancaspase inhibitor, zVAD-fmk, prevented CD95 triggered cell death in Jurkat T cells but not in A3.01 T cells, although typical hallmarks of apoptosis, such as DNA fragmentation or caspase activation were blocked. Moreover, the caspase-independent cell death in A3.01 cells exhibited typical signs of necrosis as detected by a rapid loss of cell membrane integrity and could be prevented by treatment with the radical scavenger butylated hydroxyanisole (BHA). Similar to CD95-induced cell death, apoptosis triggered by the DNA topoisomerase inhibitors, camptothecin or etoposide was shifted to necrosis when capsase activation was inhibited. In contrast to this, ZVAD was fully protective when apoptosis was triggered by the serpase inhibitor, Nalpha tosyl-phenyl-chloromethyl ketone (TPCK). TPCK was not protective when administered to anti-CD95/ZVAD-treated A3.01 cells, indicating that TPCK does not possess anti-necrotic activity but fails to activate the necrotic death pathway. Our findings show (a) that caspase inhibition does not always protect apoptotic T cells from dying but merely activates a caspase-independent mode of cell death that results in necrosis and (b) that the caspase-inhibitor-induced shift from apoptotic to necrotic cell death is dependent on the cell type and the proapoptotic stimulus. PMID- 16365882 TI - Cerebral folate deficiency and leukoencephalopathy caused by a mitochondrial DNA deletion. AB - OBJECTIVE: Our aim was to describe a child with an incomplete form of Kearns Sayre syndrome who presented profound cerebrospinal fluid (CSF) folate deficiency and his response to folinic acid supplementation METHODS: CSF 5 methyltetrahydrofolate was analyzed by HPLC with fluorescence detection and mitochondrial DNA deletions by southern blot hybridization. RESULTS: Cranial magnetic resonance imaging showed a leukoencephalopathy. Profound CSF 5 methyltetrahydrofolate deficiency was observed with normal blood folate values and decreased CSF/serum folate ratio, suggesting a transport defect across the blood-brain barrier. Folinic acid treatment was established, and after 1 year clinical response to folinic supplementation was remarkable, with almost normal white matter image. INTERPRETATION: The clinical response after folinic therapy highlights the need for the study of cerebral folate deficiency in patients with mitochondrial disorders and white matter lesions. PMID- 16365883 TI - Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells. AB - Gravity acts permanently on organisms as either static or dynamic stimulation. Understanding the influence of gravitational and mechanical stimuli on biological systems is an intriguing scientific problem. More than two decades of life science studies in low g, either real or modeled by clinostats, as well as experimentation with devices simulating different types of controlled mechanical stimuli, have shown that important biological functions are altered at the single cell level. Here, we show that the human leukemic line FLG 29.1, characterized as an osteoclastic precursor model, is directly sensitive to gravitational unloading, modeled by a random positioning machine (RPM). The phenotypic expression of cytoskeletal proteins, osteoclastic markers, and factors regulating apoptosis was investigated using histochemical and immunohistochemical methods, while the expression of the corresponding genes was analyzed using RT-PCR. A quantitative bone resorption assay was performed. Autofluorescence spectroscopy and imaging were applied to gain information on cell metabolism. The results show that modeled hypogravity may trigger both differentiation and apoptosis in FLG 29.1 cells. Indeed, when comparing RPM versus 1 x g cultures, in the former we found cytoskeletal alterations and a marked increase in apoptosis, but the surviving cells showed an osteoclastic-like morphology, overexpression of osteoclastic markers and the ability to resorb bone. In particular, the overexpression of both RANK and its ligand RANKL, maintained even after return to 1 x g conditions, is consistent with the firing of a differentiation process via a paracrine/autocrine mechanism. PMID- 16365884 TI - Light-induced exocytosis in cell development and differentiation. AB - Calcium-dependent exocytosis of fluorescently labeled single secretory vesicles in PC12 cells and primary embryonic telencephalon cells can be triggered by illumination with visible light and imaged by TIRF or epifluorescence microscopy. Opsin 3 was identified by quantitative PCR expression analysis as the putative light receptor molecule for light-induced exocytosis. In primary chicken telencephalon cells, light-induced exocytosis is restricted to a specific period during embryonic development, and involves fusion of rather large vesicles. Strictly calcium-dependent exocytosis starts after a delay of a few seconds of illumination and lasts for up to 2 min. We analyzed the frequency, time course and spatial distribution of exocytotic events. Exocytosis in PC12 cells and telencephalon cells occurs at the periphery or the interface between dividing cells, and the duration of single secretion events varies considerably. Our observation strongly supports the idea that light induced exocytosis is most likely a mechanism for building plasma membrane during differentiation, development and proliferation rather than for calcium-dependent neurotransmitter release. PMID- 16365885 TI - Modified p27 Kip1 is efficient in suppressing HER2-mediated tumorigenicity. AB - Cyclin-dependent kinase (CDK) inhibitor p27 Kip1, a haplo-insufficient tumor suppressor, is downregulated by oncogenic signal of HER2, a receptor tyrosine kinase oncogene. HER2 promotes mitogenic growth and transformation of cancer cells. HER2 signaling can enhance p27 Kip1 ubiquitination, thereby promoting p27 degradation and subsequent activation of CDK activity. p27 ubiquitination and degradation is enhanced by JAB1 binding as well as by phosphorylation on Thr187. In this study, we generated modified p27 proteins, which are mutated at Thr 187 or deleted at JAB1 binding domain. We applied these modified p27 genes as novel anticancer agents for HER2-overexpressing cells under the control of a tetracycline (tet)-regulated gene expression system. Induction of p27 T187A and p27 T187A DeltaJAB inhibits HER2-activated cell growth, CDK2 activity, cell proliferation, and transformation. Significantly, a modified protein (p27 T187ADeltaJAB) reduced the tumor volume in a HER2-overexpressing tumor model efficiently. These findings demonstrate the applicability of employing modified p27 proteins as a therapeutic intervention in HER2-overexpressing cancers. PMID- 16365886 TI - Stokes polarimetry imaging of rat-tail tissue in a turbid medium using incident circularly polarized light. AB - BACKGROUND AND OBJECTIVES: We describe a Stokes polarimetry imaging technique that quantifies the polarization properties of remitted light backscattered from a sample. STUDY DESIGN/MATERIALS AND METHODS: Right- and left-circularly polarized near-infrared light was used to illuminate rat-tail tissue embedded in turbid gelatin. RESULTS: The degree of linear polarization (DoLP) and degree of circular polarization (DoCP) image-maps indicate that increasing the depth of the rat tail within the turbid medium and varying the rat-tail geometry and orientation relative to the light source affected the contrast between structures and adjacent tissue layers. CONCLUSION: Stokes polarimetry imaging shows that the intervertebral discs and soft tissue regions of rat tails strongly depolarize incident circularly polarized light. Tendon regions remit light with a more linear form due to birefringence. Both DoLP and DoCP image-maps provide contrast between tissue structures. When differentiating between unpolarized light and light with low DoCP or DoLP, the polarization of backscattered light from the turbid medium must to be taken into consideration. PMID- 16365887 TI - Treatment of hypertrophic scars and keloids with a radiofrequency device: a study of collagen effects. AB - BACKGROUND AND OBJECTIVE: To determine the efficacy and safety of the ThermaCool TC radiofrequency system for treatment of hypertrophic and keloid scars and evaluate treatment associated collagen changes. MATERIALS AND METHODS: Six subjects with hypertrophic and four with keloid scars were treated with the ThermaCool device: one-third of the scar received no treatment (control), one third received one treatment and one-third received two treatments (4-week interval). Scars were graded before and then 12 and 24 weeks after treatment on symptoms, pigmentation, vascularity, pliability, and height. Biopsies were taken from four subjects with hypertrophic scars and evaluated with hematoxylin and eosin (H & E) staining, multiphoton microscopy, and pro-collagen I and III immunohistochemistry. RESULTS: No adverse treatment effects occurred. Clinical and H & E evaluation revealed no significant differences between control and treatment sites. Differences in collagen morphology were detected in some subjects. Increased collagen production (type III > type I) was observed, appeared to peak between 6 and 10 weeks post-treatment and had not returned to baseline even after 12 weeks. CONCLUSION: Use of the thermage radiofrequency device on hypertrophic scars resulted in collagen fibril morphology and production changes. ThermaCool alone did not achieve clinical hypertrophic scar or keloid improvement. The collagen effects of this device should be evaluated further in order to optimize its therapeutic potential for all indications. PMID- 16365888 TI - Porcine skin ED50 damage thresholds for 2,000 nm laser irradiation. AB - BACKGROUND AND OBJECTIVES: To gain refinement in safe-exposure limits, indicated by the maximum permissible exposure (MPE) limits, the minimum visible lesion thresholds for three spot sizes (5-15 mm) and four exposure durations (0.25-2.5 seconds) were determined for the skin at 2,000 nm continuous wave laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: A series of experiments were conducted in vivo on female Yucatan mini-pigs to determine the ED50 damage thresholds for 2,000 nm continuous wave laser irradiation. The study employed Gaussian laser beam exposures with spot diameters (1/e2) of 4.83, 9.65, and 14.65 mm and exposure durations of 0.25, 0.5, 1.0, and 2.5 seconds as a function of laser power. The effect of each irradiation was evaluated within 1 minute after irradiation and the final determination was made at 48 hours post-exposure. Probit analysis was conducted to estimate the dose for 50% probability of laser induced damage (ED50), defined as persistent redness at the site of irradiation for the mini-pig skin after 48 hours. RESULTS: The MPE spot size and exposure duration trends for 2,000 nm laser exposure is consistent for exposure diameters less than 3.5 mm. However, for larger exposure diameters of 4.83, 9.65, and 14.65 mm and exposure duration longer than 0.25 second, the current MPEs are bigger than one tenth of our damage thresholds. For Gaussian laser profile, which is common for many laser output irradiance distributions, lower energy is required to generate a lesion on skin for smaller spot sizes and shorter exposure duration. On the other hand, for spot sizes greater than 4.83 mm and exposure duration over 0.25 second, the average radiant exposure at threshold is inversely proportional to spot size. The irradiance-time and temperature-time power law at the threshold were investigated as well and showed that the irradiance-time power law was a close approximation to estimate laser irradiance at ED50 damage threshold. CONCLUSIONS: The thresholds study shows that consideration for lowering the MPE standards should be explored as the laser beam diameter becomes larger than 3.5 mm. Based on the limited experimental data, the duration and size dependences of the ED50 damage thresholds could be described by an empirical equation: Irradiance at the threshold = (5.669-1.81xspot diameter)xexposure duration -0.794. PMID- 16365889 TI - Use of lasers and light-based therapies for treatment of acne vulgaris. AB - Over the last two decades, lasers and light-based therapies have been developed to treat a wide variety of cutaneous maladies. Given the prevalence and number of patients who suffer from refractory acne, alternatives to existing care are constantly sought after. In this review, we discuss the evidence currently available to justify the use of laser and light-based modalities and conclude that in combination therapy, such approaches provide a safe and effective treatment for acne vulgaris. PMID- 16365890 TI - Clinical efficacy of semiconductor laser application as an adjunct to conventional scaling and root planing. AB - BACKGROUND AND OBJECTIVES: The aim of the in vitro study was to examine the clinical efficacy of semiconductor laser periodontal pocket irradiation as an adjunct to conventional scaling and root planing. MATERIALS AND METHODS: Twenty two healthy patients with a need of periodontal treatment (15 women, 7 men, mean age 45.0 +/- 10.8 years) with at least four teeth in all quadrants, were included. All of them underwent a conventional periodontal treatment including scaling and root planing. Using a split mouth design, two randomly chosen quadrants (one upper and the corresponding lower one) were subsequently treated with an 809 nm GaAlAs laser operated at a power output of 1.0 Watt using a 0.6 mm optical fiber. The teeth in the control quadrants were rinsed with saline. The clinical outcome was evaluated by means of plaque index (PI), gingival index (GI), bleeding on probing (BOP), sulcus fluid flow rate (SFFR), Periotest (PT), probing pocket depth (PPD), and clinical attachment loss (CAL) at baseline and at 3 months after treatment. A total of 492 teeth in both groups were evaluated and differences between the laser and the control teeth were analyzed using the Wilcoxon test (P < 0.05). RESULTS: Teeth treated with the laser revealed a significantly higher reduction in tooth mobility, pocket depth, and clinical attachment loss. Twelve percent of the teeth in the laser group showed an attachment gain of 3 mm or more, compared to 7% in the control group. An attachment gain of 2-3 mm was found in 24% of the teeth in the laser group and 18% in the control group. No significant group differences, however, could be detected for the plaque index, gingival index, bleeding on probing, and the sulcus fluid flow rate. CONCLUSIONS: The higher reduction in tooth mobility and probing depths is probably not predominantly related to bacterial reduction in the periodontal pockets but to the de-epithelization of the periodontal pockets leading to an enhanced connective tissue attachment. The application of the diode laser in the treatment of inflammatory periodontitis at the irradiation parameters described above is a safe clinical procedure and can be recommended as an adjunct to conventional scaling and root planing. PMID- 16365892 TI - PGF2alpha increases FGF-2 and FGFR2 trafficking in Py1a rat osteoblasts via clathrin independent and importin beta dependent pathway. AB - Previous studies showed that prostaglandin F2alpha (PGF2alpha) stimulated fibroblast growth factor-2 (FGF-2) and fibroblast growth factor receptor 2 (FGFR2) cytosolic and nuclear accumulation, however, the endocytic pathway has not been elucidated. This study demonstrates that although PGF2alpha increased the formation of clathrin-coated structures in Py1a rat osteoblasts, they were not involved in FGF-2 and FGFR2 trafficking. PGF2alpha increased binding of FGF-2 and FGFR2 and co-localization of reactive sites in addition to nuclear translocation at the nuclear pore complex level. FGF-2 and FGFR2 were in close spatial correlation with importin beta, further supporting nuclear import of the FGF-2/FGFR2 complex. Immunogold and immunofluorescence techniques as well as Western blotting demonstrated increased importin beta protein labeling in response to PGF2alpha. Similar to PGF2alpha, phorbol 12-myristate 13-acetate (PMA) also increased importin beta protein. These data strongly suggest that prostaglandins may regulate osteoblast metabolism via FGF-2/FGFR2/importin beta nuclear trafficking. PMID- 16365893 TI - Caspase-dependent and -independent cell death induced by 3-nitropropionic acid in rat cortical neurons. AB - Mitochondria play a critical role in cell death by releasing apoptogenic factors, such as cytochrome c and apoptosis-inducing factor (AIF), from the intermembrane space into the cytoplasm. Because mitochondrial dysfunction has been shown to be involved in several neurodegenerative diseases, mitochondrial toxins are largely used to model these disorders. These include 3-nitropropionic acid (3-NP), an irreversible inhibitor of succinate dehydrogenase, which has been used to model Huntington's disease and was previously reported by us to induce apoptotic cell death through caspase activation. In the present study, we evaluated the involvement of caspase-independent neuronal cell death induced by 3-NP (1 mM) and the effect of z-VDVAD-fmk, an inhibitor of caspase-2, using cortical neurons in culture. Our results highly suggest that 3-NP induces both caspase-dependent and independent cell death. We showed that z-VDVAD-fmk prevented both caspase-2 and 3-like activities evoked by 3-NP, but only partly prevented chromatin fragmentation/condensation. However, z-VDVAD-fmk did not avoid 3-NP-induced release of cytochrome c or AIF from mitochondria nor did it affect the levels of mitochondrial Bax. Furthermore, 3-NP-mediated decrease in plasma membrane integrity was not affected by z-VDVAD-fmk. Under these conditions, the inhibitor prevented the caspase-dependent cell death. PMID- 16365895 TI - Expression of LINE-1 retroposons is essential for murine preimplantation development. AB - In higher eukaryotes, reverse transcriptase (RT) activities are encoded by a variety of endogenous retroviruses and retrotransposable elements. We previously found that mouse preimplantation embryos are endowed with an endogenous RT activity. Inhibition of that activity by the non nucleosidic inhibitor nevirapine or by microinjection of anti-RT antibody caused early embryonic developmental arrest. Those experiments indicated that RT is required for early development, but did not identify the responsible coding elements. We now show that microinjection of morpholino-modified antisense oligonucleotides targeting the 5' end region of active LINE-1 retrotransposons in murine zygotes irreversibly arrests preimplantation development at the two- and four-cell stages; the overall level of functional RT is concomitantly downregulated in arrested embryos. Furthermore, we show that the induction of embryo developmental arrest is associated with a substantial reprogramming of gene expression. Together, these results support the conclusion that expression of LINE-1 retrotransposons is required for early embryo preimplantation development. PMID- 16365894 TI - RANK ligand expression in heat shock factor-2 deficient mouse bone marrow stromal/preosteoblast cells. AB - Heat Shock Proteins (HSP) are molecular chaperones activated upon cellular stress/stimuli. HSP gene expression is regulated by Heat Shock Factors (HSF). We have recently demonstrated a functional role for heat shock factor-2 (HSF-2) in fibroblast growth factor-2 (FGF-2)-induced RANK ligand (RANKL), a critical osteoclastogenic factor expression on stromal/preosteoblast cells. In the present study, we show that FGF-2 treatment did not induce RANKL expression in HSF-2-/ stromal/preosteoblast cells. Interestingly, HSF-2 deficiency resulted in rapid induction of alkaline phosphatase (ALP) activity and osteocalcin mRNA expression in these cells. Furthermore, FGF-2 did not induce osteoclast formation in co culture of normal mouse spleen cells and HSF-2-/-stromal/preosteoblast cells. Electron microscopy analysis demonstrated that osteoclasts from HSF-2-/-mice have poorly developed ruffled borders. These data further confirm that HSF-2 plays an important role in FGF-2-induced RANKL expression in stromal/preosteoblast cells. HSF-2 deficiency has pleotropic effects on gene expression during osteoblast differentiation and osteoclastogenesis in the bone microenvironment. Novel therapeutic agents that modulate HSF-2 activation may have therapeutic utility against increased levels of FGF-2 and bone destruction associated with pathologic conditions. PMID- 16365896 TI - Sharing the diagnostic process in the clinical teaching environment: a case study. AB - Revealing or visualizing the thinking involved in making clinical decisions is a challenge. A case study is presented with a visual implement for sharing the diagnostic process. This technique adapts the Bayesian approach to the case presentation. Pretest probabilities and likelihood ratios are gathered to obtain post-test probabilities of every question considered in a patient. The technique advocates for the judicious use of diagnostic options based on the best evidence available. Clinical decisions, presentation skills, and critical evaluation of the literature are also important elements considered in this method. PMID- 16365897 TI - Design and implementation of training to improve management of pediatric overweight. AB - INTRODUCTION: Clinicians report a low proficiency in treating overweight children and using behavioral management strategies. This paper documents the design and implementation of a training program to improve clinicians' skills in the assessment and behavioral management of pediatric overweight. METHODS: Two one hour CME trainings were designed using published guidelines, research findings, and expert committee recommendations. The trainings were provided to clinicians of a managed care pediatric department, utilizing novel screening and counseling tools, and interactive exercises. Surveys and focus groups were conducted 3 and 6 months post intervention to examine clinician attitudes and practices regarding the screening and counseling tools. RESULTS: Post intervention, the majority of clinicians agreed that the clinical practice guidelines (Pediatric Obesity Practice Resource) and BMI-for-age percentile provided useful information for clinical practice. Clinicians reported an increased utilization of the recommended screening tools and changes in office practices to implement these tools. They offered suggestions to improve the ease of use of the tools and to overcome perceived clinician and/or patient barriers. DISCUSSION: A brief, cost effective, multi-faceted training and provision of counseling tools were perceived as helpful to clinical practice. Useful lessons were learned about tool design and ways to fit tools into practice. Training the entire health care team is advantageous to the adoption of new tools and practices. PMID- 16365898 TI - Contents of a core library in continuing medical education: a Delphi study. AB - INTRODUCTION: In developing their professional competence, those who are interested in the practice of continuing medical education (CME) should recognize the knowledge base that defines their field. This study systematically identifies and organizes a list of books and journals comprising a core library (100 books/15 journals) for CME professionals. METHODS: The Delphi method was applied to elicit and combine the judgments of a fifty member panel considered knowledgeable about the CME field. The panelists participated in three iterations of the survey to first identify and then rank order nominated works. Separate ranked lists were created for books and journals. RESULTS: Forty-four participants completed the study (88% response rate). 268 books and 34 journals were identified. Mean ratings ranged from 4.78 (high) to 1.50 (low). DISCUSSION: The results of the study reflect the panel's judgment. The list is not definitive; instead, it describes what a select group of individuals knowledgeable about the CME field considered important. The list should therefore be seen as a general guide and a resource to facilitate decision-making, not as a prescription for creating a library. PMID- 16365899 TI - Defining and assessing the competence of health care professionals in France. AB - INTRODUCTION: During the past decade, the public, health care professionals, and governments have shown much interest in ensuring that professionals, specifically physicians, are skilled in their work and competent in managing health care organizations. The need to assess competence in the health care sector was explored, with the aim of proposing a policy for monitoring and assessing competence during active professional life. METHODS: A literature search and semistructured interviews were conducted. The 16 health care professions listed in the French health care code were included. The main questions were: What is your organization's definition of competence? What are the principal elements that define competence in health care activities? How can a system for assessing competence be implemented? Which methods for such a system are most appropriate, based on foreign experiences? Who are the players in the field of competence? And how can organizations participate in monitoring competence? RESULTS: 265 people representing 148 French organizations were interviewed. Competence in health care should be defined as follows: "professional competence is based on the initial diploma, the implementation of effective continuing education, a minimal professional activity and a regular peer review process." There was an agreement on the basic elements of competence, on the responsibilities of public institutions and professional organizations, and on the need to work together. DISCUSSION: We have shown that in France health care professionals would like to have a better system that allows them to exchange more information on the main health care issues; this is a serious requirement for most professionals. PMID- 16365900 TI - Effect of CME on primary care and OB/GYN treatment of breast masses. AB - INTRODUCTION: CME program planners are being asked to move beyond assessments of knowledge to assessing the impact of CME on practice and patient outcomes. METHODS: We conducted a pre-post analysis of administrative data from 107 physicians, nurse practitioners (NPs), or physician's assistants (PAs) who attended one or two continuing medical education (CME) programs (an in-person, mainly didactic session on breast complaints in women, or an individual mentorship with general surgeons) between August 2002 and March 2003. We examined associations between the number of trainings and attempted breast mass aspirations or general surgery referrals for breast masses; individual training and breast mass aspiration attempts or general surgery referrals; and provider type and attempted breast mass aspirations. Generalized linear mixed models were used to model dichotomous outcomes. RESULTS: Clinicians who participated in individual trainings performed more breast mass aspirations after training (odds ratio (OR) 3.07, [95% confidence interval 1.10-8.54]). Participants who completed two trainings performed more breast mass aspirations after training (OR 2.33, [1.19-4.57]), while those who completed just one did not (OR 1.34, [0.39, 4.58]) but the effect started after the first training and did not strengthen after the second training. NPs and PAs attempted more aspirations after training (OR 6.1, [1.54, 24.1]), whereas physicians did not (OR 0.89 [0.36, 2.22]). Training was not associated with a change in referrals to general surgery. Referral appropriateness, pre-training readiness to change, and previous training in breast mass aspiration were not assessed. DISCUSSION: Attempts to aspirate breast masses may increase after CME training. Individual training may be more effective than group training in increasing the likelihood of attempted aspirations. PMID- 16365901 TI - Evaluation of a training to improve management of pediatric overweight. AB - INTRODUCTION: Despite widespread concern about pediatric obesity, health care professionals report low proficiency for identifying and treating this condition. This paper reports on the evaluation of pediatric overweight assessment and management training for clinicians and staff in a managed care system. The training was evaluated for its impact on assessment practices and utilization of management tools. METHODS: A delayed-control design was utilized to measure the effects of two 60-minute interactive Continuing Medical education (CME) trainings for the pediatric health care teams. Chart abstraction was conducted at 0-, 3- and 6-months after training, recording the proportion of charts containing the recommended assessment methods and management tools. RESULTS: The training was associated with a significant increase in the utilization of some tools and practices, including charting BMI-for-age percentile (p < 0.001) and using a nutrition and activity self-history form (p < 0.001). Overall, from baseline to 3 months post training, charting BMI-for-age percentiles increased from zero to 25.2% and utilization of the self-history form increased from zero to 35.3%. These increases were sustained at 6-months post training. Other tools guiding clinician counseling were less widely utilized, although a behavioral prescription pad was used with 20% of overweight patients. DISCUSSION: A modest investment in clinician and staff training designed to be feasible in a clinical setting was associated with substantial increases in the use of appropriate tools and practices for the assessment and management of pediatric overweight. Such training may help to augment and improve the processes of pediatric health care delivery for addressing overweight. The training provides a viable model for future CME efforts in other health care settings. PMID- 16365902 TI - Continuing professional development: racial and gender differences in obstetrics and gynecology residents' perceptions of mentoring. AB - INTRODUCTION: Having a mentor during residency is often linked to greater success in professional development. The present study compares changes in the percentage of residents with mentors in 1999 and in 2004, while considering current residents' perceptions of their mentors, with particular attention focused on what role race and gender might play in resident-mentor interactions. METHOD: A survey was administered to 4, 721 residents who took the 2004 Council on Resident Education in Obstetrics and Gynecology in-training examination. Data are reported for respondents from four racial categories: white, African American, Hispanic, and Asian/Pacific Islander. Results were compared to those of a similar survey administered in 1999. Responses were analyzed by chi-square analysis and univariate analyses of variance. RESULTS: The response rate was 97%. Most residents (64.9%) reported having a mentor. White female residents were least likely to have a mentor. Compared to results from 1999, the percentage of residents with a mentor, and the percentage of residents with female mentors, has increased. For all residents, personal rapport, knowledge of the field, and similarity in professional interests were the three most important factors in choosing a mentor. The proportion of residents reporting explicit discussions about career options has declined since 1999. DISCUSSION: Ethnic minorities are more likely than white residents to have a mentor, and to report that their mentors provide helpful advice. Although the proportion of residents with a mentor has increased since 1999, the quality of the mentoring relationship is meeting resident expectations but not exceeding them. PMID- 16365904 TI - Concise enantio- and diastereoselective total syntheses of fumagillol, RK-805, FR65814, ovalicin, and 5-demethylovalicin. PMID- 16365905 TI - Oligo-(1-->2)-alpha-D-glucopyranosyl-(1-->2)-beta-D-fructofuranosides form tight sugar coils. PMID- 16365906 TI - Widely applicable Pd-catalyzed trans-selective monoalkylation of unactivated 1,1 dichloro-1-alkenes and Pd-catalyzed second substitution for the selective synthesis of E or Z trisubstituted alkenes. PMID- 16365907 TI - Design, synthesis, and preliminary biological evaluation of a DNA methyltransferase-directed alkylating agent. PMID- 16365908 TI - Cationic P-S-X cages (X=Br, I). AB - The first condensed-phase preparation of ternary P-Ch-X cations (Ch=O-Te, X=F-I) is reported: [P5S3X2]+, [P5S2X2]+, and [P4S4X]+ (X=Br, I). [P5S3X2]+ is formed from the reaction of the Ag+/PX3 reagent with P4S3. The [P5S3X2]+ ions have a structure that is related to P4S5 by replacing P=S by P+--X and S in the four membered ring by P(X). We provide evidence that the active ingredient of the Ag+/PX3 reagent is the (H2CCl2)Ag-X-PX2+ cation. The latter likely reacts with the HOMO of P4S3 in a concerted HOMO-LUMO addition to give the P5S3X2+ ion as the first species visible in situ in the low-temperature 31P NMR spectrum. The [P5S3X2]+ ions are metastable at -78 degrees C and disproportionate at slightly higher temperatures to give [P5S2X2]+ and [P4S4X]+, probably with the extrusion of 1/n (PX)n (X=Br, I). All six new cage compounds have been characterized by multinuclear NMR spectroscopy and, in part, by IR or Raman spectroscopy. The [P5S2X2]+ salts have a nortricyclane skeleton and were also characterized by X ray crystallography. The structure of the [P4S4X]+ ion is related to that of P4S5 in that the exo-cage P=S bond is replaced by an isoelectronic P+--X moiety. PMID- 16365910 TI - Discounting and cost-effectiveness in NICE - stepping back to sort out a confusion. AB - Brouwer and colleagues [1] argue that the reasons for specifying an equal discount rate for health outcomes and costs in the recent guidance on methods of technology appraisal issued by the National Institute for Clinical Excellence (NICE) [2] is both opaque and wrong. They argue that a lower rate should apply to health outcomes like QALYs. It is also claimed that the guidance on discounting represents a step backwards, that is both inconsistent with current theoretical insights and will prejudice the outcome of cost-effectiveness studies of preventive interventions.The reasoning behind the use of equal discount rates for costs and health outcomes is indeed not well developed in the published guidance. Nor does it reflect the debate that underpinned the guidance. We therefore welcome the opportunity to explain more completely the rationale in the minds of the principal authors of the current guidance. PMID- 16365912 TI - Conformational analysis of a potent SSTR3-selective somatostatin analogue by NMR in water solution. AB - The three-dimensional structure of a potent SSTR3-selective analogue of somatostatin, cyclo(3-14)H-Cys(3)-Phe(6)-Tyr(7)-D-Agl(8)(N(beta) Me, 2-naphthoyl) Lys(9)-Thr(10)-Phe(11)-Cys(14)-OH (des-AA(1, 2, 4, 5, 12, 13)[Tyr(7), D Agl(8)(N(beta) Me, 2-naphthoyl)]-SRIF) (peptide 1) has been determined by (1)H NMR in water and molecular dynamics (MD) simulations. The peptide exists in two conformational isomers differing mainly by the cis/trans isomerization of the side chain in residue 8. The structure of 1 is compared with the consensus structural motifs of other somatostatin analogues that bind predominantly to SSTR1, SSTR2/SSTR5 and SSTR4 receptors, and to the 3D structure of a non selective SRIF analogue, cyclo(3-14)H-Cys(3)-Phe(6)-Tyr(7)-D-2Nal(8)-Lys(9) Thr(10)-Phe(11)-Cys(14)-OH (des-AA(1, 2, 4, 5, 12, 13)[Tyr(7), D-2Nal(8)]-SRIF) (peptide 2). The structural determinant factors that could explain selectivity of peptide 1 for SSTR3 receptors are discussed. PMID- 16365913 TI - Regulation of T cells and cytokines by the interleukin-10 (IL-10)-family cytokines IL-19, IL-20, IL-22, IL-24 and IL-26. AB - The family of IL-10-related cytokines includes several human members, IL-19, IL 20, IL-22, IL-24 and IL-26, and a series of herpesviral and poxviral paralogs. Some of these cytokines share common receptor subunits. In this study, we investigated the effects of these cytokines on naive T cell differentiation, antigen-specific T cell suppression, survival ad expression of surface markers in comparison to IL-10 and cytomegalovirus (CMV)-IL-10. Human CD45RA(+) T cells were stimulated in the presence of IL-10-family cytokines in sequential 12-day cycles. After three to four cycles of stimulation, IL-10 and CMV-IL-10 led to increased IFN-gamma and IL-10 but decreased IL-4 and IL-13. Interestingly, long-term exposure of T cells to IL-19, IL-20 and IL-22 down-regulated IFN-gamma but up regulated IL-4 and IL-13 in T cells and supported the polarization of naive T cells to Th2-like cells. In contrast, neutralization of endogenous IL-22 activity by IL-22-binding protein decreased IL-4, IL-13 and IFN-gamma synthesis. The antigen-specific suppressor activity of IL-10 and CMV-IL-10 was not observed for any of the other IL-10-family cytokines. These data demonstrate that IL-19, IL-20 and IL-22 may participate in T cell-mediated diseases by distinct regulation of T cell cytokine profiles. PMID- 16365915 TI - Alpha-stabilization of carbanions: fluorine is more effective than the heavier halogens. PMID- 16365914 TI - A novel lipid A from Halomonas magadiensis inhibits enteric LPS-induced human monocyte activation. AB - Lipopolysaccharide (LPS) endotoxin is the bacterial product responsible for the clinical syndrome of Gram-negative septicemia and endotoxic shock. During sepsis, microbial antigens, such as LPS, activate monocytes and macrophages to produce several pro-inflammatory cytokines, among which tumor necrosis factor-alpha (TNF alpha) appears to be very important for the development of endotoxic shock. The endotoxic properties of LPS principally reside in the lipid A (LIP A) component, which is the primary immunostimulatory center of Gram-negative bacteria. In recent years there has been a continuous effort to identify molecules able to antagonize the deleterious effects of endotoxic shock. In this study we show that a novel LIP A fraction from the LPS of Halomonas magadiensis (Hm), a Gram negative extremophilic and alkaliphilic bacterium, significantly inhibits the synthesis of TNF-alpha by human monocytes activated by Escherichia coli LPS. LIP A from Hm exerts these effects by interfering with E. coli LPS for activation of Toll-like receptor 4 expressed in human cells. This result defines Hm LIP A as a novel class of LPS antagonist whose structural features could be utilized for the design of compounds for the treatment of Gram-negative sepsis. PMID- 16365916 TI - Chemistry of superheavy elements. AB - The number of chemical elements has increased considerably in the last few decades. Most excitingly, these heaviest, man-made elements at the far-end of the Periodic Table are located in the area of the long-awaited superheavy elements. While physical techniques currently play a leading role in these discoveries, the chemistry of superheavy elements is now beginning to be developed. Advanced and very sensitive techniques allow the chemical properties of these elusive elements to be probed. Often, less than ten short-lived atoms, chemically separated one atom-at-a-time, provide crucial information on basic chemical properties. These results place the architecture of the far-end of the Periodic Table on the test bench and probe the increasingly strong relativistic effects that influence the chemical properties there. This review is focused mainly on the experimental work on superheavy element chemistry. It contains a short contribution on relativistic theory, and some important historical and nuclear aspects. PMID- 16365917 TI - Pronounced supramolecular order in discotic donor-acceptor mixtures. PMID- 16365918 TI - EHPred: an SVM-based method for epoxide hydrolases recognition and classification. AB - A two-layer method based on support vector machines (SVMs) has been developed to distinguish epoxide hydrolases (EHs) from other enzymes and to classify its subfamilies using its primary protein sequences. SVM classifiers were built using three different feature vectors extracted from the primary sequence of EHs: the amino acid composition (AAC), the dipeptide composition (DPC), and the pseudo amino acid composition (PAAC). Validated by 5-fold cross tests, the first layer SVM classifier can differentiate EHs and non-EHs with an accuracy of 94.2% and has a Matthew's correlation coefficient (MCC) of 0.84. Using 2-fold cross validation, PAAC-based second layer SVM can further classify EH subfamilies with an overall accuracy of 90.7% and MCC of 0.87 as compared to AAC (80.0%) and DPC (84.9%). A program called EHPred has also been developed to assist readers to recognize EHs and to classify their subfamilies using primary protein sequences with greater accuracy. PMID- 16365919 TI - Heuristic algorithm for off-lattice protein folding problem. AB - Enlightened by the law of interactions among objects in the physical world, we propose a heuristic algorithm for solving the three-dimensional (3D) off-lattice protein folding problem. Based on a physical model, the problem is converted from a nonlinear constraint-satisfied problem to an unconstrained optimization problem which can be solved by the well-known gradient method. To improve the efficiency of our algorithm, a strategy was introduced to generate initial configuration. Computational results showed that this algorithm could find states with lower energy than previously proposed ground states obtained by nPERM algorithm for all chains with length ranging from 13 to 55. PMID- 16365920 TI - Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system. AB - v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl beta-D thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression. PMID- 16365921 TI - Species diversity of Lachnum (Helotiales, Hyaloscyphaceae) from temperate China. AB - Twenty-three temperate China species of Lachnum, Lachnum abnorme, L. angustum, L. brevipilosum, L. calosporum, L. calyculiforme, L. carneolum, L. ciliare, L. controversum, L. flavidulum, L. cf. fushanese, L. indicum, L. kumaonicum, L. lushanese, L. minutum, L. montanum, L. cf. pteridophyllum, L. pygmaeum, L. sclerotii var. sclerotii, L. sclerotii var. sichuanense, L. subpygmeaum, L. tenuissimum, L. virgineum and L. willisii are reported, whose main characteristics are given in a formula of the described species, some of which are discussed below. PMID- 16365922 TI - Promoter trapping in Magnaporthe grisea. AB - Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M. grisea. A library of 1,077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures of M. grisea. Three transformants grew slowly, 5 transformants decreased in conidiation and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis. PMID- 16365923 TI - Culture of Spirulina platensis in human urine for biomass production and O(2) evolution. AB - Attempts were made to culture Spirulina platensis in human urine directly to achieve biomass production and O(2) evolution, for potential application to nutrient regeneration and air revitalization in life support system. The culture results showed that Spirulina platensis grows successfully in diluted human urine, and yields maximal biomass at urine dilution ratios of 140 approximately 240. Accumulation of lipid and decreasing of protein occurred due to N deficiency. O(2) release rate of Spirulina platensis in diluted human urine was higher than that in Zarrouk medium. PMID- 16365924 TI - Study on NO(2) absorption by ascorbic acid and various chemicals. AB - Study on NO(2) absorption aimed at seeking a better NO(2) absorption chemical at pH 4.5 approximately 7.0 for application to existing wet flue gas desulfurization (FGD). The results from the double-stirred reactor indicated that ascorbic acid has very high absorption rate at this pH range. The rate constant of ascorbic acid reaction with NO(2) (0 approximately 1,000 x 10(-6) mol/mol) is about 3.54 x 10(6) mol/(Ls) at pH 5.4 approximately 6.5 at 55 degrees C. PMID- 16365925 TI - Field demonstration of reduction of lead availability in soil and cabbage (Brassica Chinensis L.) contaminated by mining tailings using phosphorus fertilizers. AB - A field demonstration of reduction of lead availability in a soil and cabbage (Brassica Chinensis L.) contaminated by mining tailings, located in Shaoxing, China was carried out to evaluate the effects of applications of phosphorus fertilizers on Pb fractionation and Pb phyto-availability in the soil. It was found that the addition of all three P fertilizers including single super phosphate (SSP), phosphate rock (PR), and calcium magnesium phosphate (CMP) significantly decreased the percentage of water-soluble and exchangeable (WE) soil Pb and then reduced the uptake of Pb, Cd, and Zn by the cabbage compared to the control (CK). The results showed that the level of 300 g P/m(2) soil was the most cost-effective application rate of P fertilizers for reducing Pb availability at the first stage of remediation, and that at this P level, the effect of WE fraction of Pb in the soil decreased by three phosphorus fertilizers followed the order: CMP (79%)>SSP (41%)>PR (23%); Effectiveness on the reduction of Pb uptake by cabbage was in the order: CMP (53%)>SSP (41%)>PR (30%). Therefore our field trial demonstrated that it was effective and feasible to reduce Pb availability in soil and cabbage contaminated by mining tailings using P fertilizers in China and PR would be a most cost-effective amendment. PMID- 16365926 TI - Nutritional quality, functional properties and anti-nutrient compositions of the larva of Cirina forda (Westwood) (Lepidoptera: Saturniidae). AB - Determination of the proximate, nutritionally valuable minerals and anti-nutrient compositions in larvae of Cirina forda (W) showed that they contained high levels of protein, (55.50%+/-1.20%) with ash, moisture, fat, and carbohydrate levels being (10.26%+/-0.01%), (10.85%+/-0.38%), (4.68%+/-0.01%) and (18.70%+/-0.84%) respectively. Fibre was not detected. Protein solubility was higher in alkaline media than in acidic media while the isoelectric points were pH 4, 6 and 9 indicating that C. forda may be useful in food formulations involving foods like meat products. Phosphorus had the highest value of (215.54+/-0.21) mg/100 g while manganese had the lowest value of (1.14+/-0.10) mg/100 g. Copper, Co, Pb, Cr and Ni were not available. Water absorption capacity was (300%+/-0.15%), oil absorption capacity was (358.44%+/-0.21%) and foaming stability was (3%+/-0.00%). The results of anti-nutritional analysis revealed that oxalate and phytic acid contents were (4.11+/-0.05) mg/100 g and (1.02+/-0.00) mg/100 g respectively and that these values fell within nutritionally accepted values. Tannin was not detected in C. forda. C. forda, a rich source of animal nutrients, usable in human diets and animal feeds formulations. PMID- 16365927 TI - Effects of two medicinal plants Psidium guajava L. (Myrtaceae) and Diospyros mespiliformis L. (Ebenaceae) leaf extracts on rat skeletal muscle cells in primary culture. AB - Crude decoction, aqueous and ethanolic extracts of two medicinal plants (Psidium guajava and Diospyros mespiliformis), widely used in the central plateau of Burkina Faso to treat many diseases were evaluated for their antagonistic effects on caffeine induced calcium release from sarcoplasmic reticulum of rat skeletal muscle cells. These different extracts showed a decrease of caffeine induced calcium release in a dose dependent manner. Comparison of the results showed that Psidium guajava leaf extracts are more active than extracts of Diospyros mespiliformis and that crude decoctions show better inhibitory activity. The observed results could explain their use as antihypertensive and antidiarrhoeal agents in traditional medicine, by inhibiting intracellular calcium release. PMID- 16365928 TI - Structural changes of oviduct of freshwater shrimp, Macrobrachium nipponense (Decapoda, Palaemonidae), during spawning. AB - The structural change of the oviduct of freshwater shrimp (Macrobrachium nipponense) during spawning was examined by electron microscopy. The oviduct wall structural characteristics seem to be influenced significantly by the spawning process. Before the parturition and ovulation, two types of epithelial cells (types I and II) are found in the epithelium. The free surfaces of type I and type II cells have very dense long microvilli. Under the type I and type II cells, are a relatively thick layer of secreting material and a layer of mostly dead cells. After ovulation, two other types of epithelial cells (types III and IV) are found in the oviduct wall epithelium. The free surface of type III cells only has short microvilli scattered on the surface. The thick layer with secreting material and the dead cell layer disappeared at this stage. In some type III cells, the leaking out of cytoplasm from broken cell membrane led to the death of these type III cells. The transformation of all four types of epithelial cells was in the order: IV-->I-->II-->III. PMID- 16365929 TI - Bread wheat varieties as influenced by different nitrogen levels. AB - Experiment was conducted to determine the effect of different nitrogen levels on four bread wheat varieties (Triticum aestivum L.) viz. Inqilab-91, Daman-98, Dera 98 and Punjab-96 at Gomal University, Dera Ismail Khan (NWFP), Pakistan during 2000 approximatey 2001. The experiment was laid out in split plot design having four replications using a net plot size of 2 m x 5 m. Nitrogen doses used were 0, 50, 100, 150 and 200 kg/ha. The results showed that different nitrogen levels had significant effects on plant height, total number of plants/m(2), number of grains/spike, number of spike/m(2), spike weight, biological yield, grain yield and grain protein content. Maximum plant height, total number of plants/m(2), number of spikes/m(2), spike weight, biological yield and grain protein content were observed at 200 kg N/ha. Among wheat varieties Daman-98 had maximum plant height, spike weight, grains/spike, 1000-grain weight, biological yield and grain yield. Inqilab-91 had heavier grains and the most grain protein content, while Dera-98 had the maximum plant population and spikes/m(2). Grain yield and biological yield were statistically similar at doses of 150 kg N/ha and 200 kg N/ha. However, dose of 200 kg N/ha, compared to dose of 150 Kg N/ha, significantly increased the protein content. PMID- 16365930 TI - Relationship between malt qualities and beta-amylase activity and protein content as affected by timing of nitrogen fertilizer application. AB - The effects of different timing of N fertilizer application at the same rate on grain beta-amylase activity, protein concentration, weight and malt quality of barley were studied. Grain beta-amylase activity and protein concentration were significantly higher in treatments where all top-dressed N fertilizer was applied at booting stage only or equally applied at two-leaf stage and booting stage than in the treatment where all top-dressed N fertilizer was applied at two-leaf age stage only. On the other hand, grain weight and malt extract decreased with increased N application at booting stage. There were obvious differences between barley varieties and experimental years in the grain and malt quality response to the timing of N fertilizer application. It was found that grain protein concentration was significantly and positively correlated with beta-amylase activity, but significantly and negatively correlated with malt extract and Kolbach index. The effect of grain protein concentration on malt quality was predominant over the effect of grain beta-amylase activity. PMID- 16365931 TI - ABCD predicts the future for mini-strokes. PMID- 16365932 TI - Nuclear stress test may stress nuclear detectors. PMID- 16365933 TI - Special coverage of the 43rd IDSA meeting. Study finds negative impact of smoking on HIV infection and immune activation. AB - Research dating from the late 1990's has found that abnormal immune activation can lead to lipodystrophy syndrome, which now is broken down to lipoatrophy and lipohypertrophy, an investigator says. PMID- 16365934 TI - FDA notifications. FDA approves pediatric dosing for ritonavir. PMID- 16365935 TI - Thinking outside the cell. PMID- 16365936 TI - Paget's disease and bisphosphonates. PMID- 16365937 TI - Anonymity vanishes for sperm donors. PMID- 16365938 TI - Hong Kong battens down the hatches. PMID- 16365939 TI - Reconstructing Afghanistan's health system. PMID- 16365940 TI - Frustration over science at the policy high table. PMID- 16365941 TI - Biologists' alarm bells ring louder. PMID- 16365942 TI - Human genome carve-up continues. PMID- 16365943 TI - Repetition priming: Is music special? AB - Using short and long contexts, the present study investigated musical priming effects that are based on chord repetition and harmonic relatedness. A musical target (a chord) was preceded by either an identical prime or a different but harmonically related prime. In contrast to words, pictures, and environmental sounds, chord processing was not facilitated by repetition. Experiments 1 and 2 using single-chord primes showed either no significant difference between chord repetition and harmonic relatedness or facilitated processing for harmonically related targets. Experiment 3 using longer prime contexts showed that musical priming depended more on the musical function of the target in the preceding context than on target repetition. The effect of musical function was decreased, but not qualitatively changed, by chord repetition. The outcome of this study challenges predictions of sensory approaches and supports a cognitive approach of musical priming. PMID- 16365944 TI - Correspondences between microcalcification projections on two mammographic views acquired with digital systems. AB - In this paper, we have proposed an algorithm for automatic matching of MC projections viewed on two mammograms of the same breast. The implemented algorithm consists in three steps. From five morphological features of the MC, a similarity function was built between each MC of the first image of the pair, and each MC of the second image. These values quantified the resemblance between each pair of MC and permitted, for a given MC of the first image, to sort the MC of the second image which could be matched to it. From the geometry of the system providing the pairs of images being analysed, we derived some geometrical constraints that must be satisfied by corresponding MC. In order to take into account the fact that due to breast deformation, the corresponding MC is often off the classical epipolar line constructed on the basis of stereovision assumption, we instead consider an epipolar strip on both side of the approximated epipolar line. The MC of the second image which was out of that strip was eliminated. Then a coefficient was applied to the remaining MC that took into account their distances to the approximated epipolar line. Finally, the selection procedure was used to pick out the right pair. In order to test our algorithm, we compared the result it yielded with those coming from two operators who matched a number of MC with confidence. A concordance of 78.43% was obtained between confident manual matching and automatic matching. Since this algorithm was designed to be part of a tool for 3D reconstruction of microcalcification clusters, we reconstructed some clusters with manual matching and automatic matching and compared the shapes of the clusters obtained in these two ways. The resemblance in some cases was very good and average in a number of others. This suggests that our algorithm should be improved. Therefore, apart from ongoing effort to introduce other constraints, we believe that taking into account the third view provided by the imaging system could be of great help. We shall soon explore this possibility. Overall, we believe that despite the failure of our tool in some cases it can already at this stage be used with some confidence. PMID- 16365945 TI - When hearing turns into playing: movement induction by auditory stimuli in pianists. AB - In this study, pianists were tested for learned associations between actions (movements on the piano) and their perceivable sensory effects (piano tones). Actions were examined that required the playing of two-tone sequences (intervals) in a four-choice paradigm. In Experiment 1, the intervals to be played were denoted by visual note stimuli. Concurrently with these imperative stimuli, task irrelevant auditory distractor intervals were presented ("potential" action effects, congruent or incongruent). In Experiment 2, imperative stimuli were coloured squares, in order to exclude possible influences of spatial relationships of notes, responses, and auditory stimuli. In both experiments responses in the incongruent conditions were slower than those in the congruent conditions. Also, heard intervals actually "induced" false responses. The reaction time effects were more pronounced in Experiment 2. In nonmusicians (Experiment 3), no evidence for interference could be observed. Thus, our results show that in expert pianists potential action effects are able to induce corresponding actions, which demonstrates the existence of acquired action-effect associations in pianists. PMID- 16365946 TI - What makes a route appear longer? An experimental perspective on features, route segmentation, and distance knowledge. AB - Five experiments performed in a desktop virtual-reality setting investigated the influence of environmental features--that is, noticeable landmarks along the route--on distance estimation. Landmarks were of two types: Either they simply "filled" the route or they "filled" and also segmented it, thereby inducing a hierarchical structuring of the route. Previous research had left the question open of whether a filling or a segmenting feature leads to an overestimation of a distance along the route. Our experiments showed different results dependent on the kind of space: If an environment was learned from a route perspective, filling and segmenting environmental features led to overestimations of distances, while the segmenting of a route induced by a grouping of similar features did not. If the environment was learned from a map that afforded a survey perspective, route structuring induced through a segmenting feature or by phenomenal grouping led to an overestimation of distances, whereas features that merely filled the route did not. PMID- 16365947 TI - Lag-1 sparing in the attentional blink: benefits and costs of integrating two events into a single episode. AB - When people monitor a visual stream of rapidly presented stimuli for two targets (T1 and T2), they often miss T2 if it falls into a time window of about half a second after T1 onset--the attentional blink. However, if T2 immediately follows T1, performance is often reported being as good as that at long lags--the so called Lag-1 sparing effect. Two experiments investigated the mechanisms underlying this effect. Experiment 1 showed that, at Lag 1, requiring subjects to correctly report both identity and temporal order of targets produces relatively good performance on T2 but relatively bad performance on T1. Experiment 2 confirmed that subjects often confuse target order at short lags, especially if the two targets are equally easy to discriminate. Results suggest that, if two targets appear in close succession, they compete for attentional resources. If the two competitors are of unequal strength the stronger one is more likely to win and be reported at the expense of the other. If the two are equally strong, however, they will often be integrated into the same attentional episode and thus get both access to attentional resources. But this comes with a cost, as it eliminates information about the targets' temporal order. PMID- 16365948 TI - Stimulus similarity decrements in children's working memory span. AB - Two experiments investigated the impact of the relationship between processing and storage stimuli on the working memory span task performance of children aged 7 and 9 years of age. In Experiment 1, two types of span task were administered (sentence span and operation span), and participants were required to recall either the products of the processing task (sentence-final word, arithmetic total) or a word or digit unrelated to the processing task. Experiment 2 contrasted sentence span and operation span combined with storage of either words or digits, in tasks in which the item to be remembered was not a direct product of the processing task in either condition. In both experiments, memory span was significantly greater when the items to be recalled belonged to a different stimulus category from the material that was processed, so that in sentence span tasks, number recall was superior to word recall, and in operation span tasks, word recall was superior to number recall. Explanations of these findings in terms of similarity-based interference and response competition in working memory are discussed. PMID- 16365949 TI - By which name should I call thee? The consequences of having multiple names. AB - The nominal competitor effect suggests that, when a person has two names associated with them, recall of either name is more difficult than if they just had one name. Drawing on a connectionist framework, this effect could arise either if multiple names were represented as being connected to a single person identity node (PIN), or if multiple names were represented as being connected via one-to-one links to multiple PINs. Whilst the latter has intuitive appeal, results from two experiments support the former architecture. Having two names connected to a single PIN not only gives rise to a nominal competitor effect (Experiment 1), but also gives rise to a familiarity enhancement effect (Experiment 2). These empirical results are simulated using an extension of Bredart, Valentine, Calder, and Gassi's (1995) connectionist architecture, which reveals that both effects hold even when the association of both names to the PIN is unequal. These results are presented in terms of a more complete model for person recognition, and the representation of semantic information within such a model is examined. PMID- 16365950 TI - Phonological similarity effects in verbal complex span. AB - Phonological similarity effects were used to assess the role of acoustic coding in verbal complex span, a processing-plus-storage measure found to correlate significantly with aspects of complex cognition. Three experiments demonstrated consistent effects of phonological similarity on listening span. These effects appeared relatively insensitive to manipulations of task materials (Experiment 1) and differences in processing task demands (Experiments 2 and 3). The results were interpreted as reflecting a significant role for the phonological loop in supporting verbal complex span and a multicomponent view of working memory, as tapped by these tests. Phonological similarity did not significantly interact with aspects of the tasks varied across Experiments 1 to 3, suggesting a relative robustness of the effect. However, variation in the phonological similarity effect sizes across Experiments 1 to 3 supports the suggestion that task demands and characteristics have the potential to disrupt the phonological similarity effect and, by implication, the reliance on a phonological code. PMID- 16365951 TI - Causal and noncausal conditionals: an integrated model of interpretation and reasoning. AB - We present an integrated model for the understanding of and the reasoning from conditional statements. Central assumptions from several approaches are integrated into a causal path model. According to the model, the cognitive availability of exceptions to a conditional reduces the subjective conditional probability of the consequent, given the antecedent. This conditional probability determines people's degree of belief in the conditional, which in turn affects their willingness to accept logically valid inferences. In addition to this indirect pathway, the model contains a direct pathway: Availability of exceptional situations directly reduces the endorsement of valid inferences. We tested the integrated model with three experiments using conditional statements embedded in pseudonaturalistic cover stories. An explicitly mentioned causal link between antecedent and consequent was either present (causal conditionals) or absent (arbitrary conditionals). The model was supported for the causal but not for the arbitrary conditional statements. PMID- 16365952 TI - Relations between emotion, illusory word perception, and orthographic repetition blindness: tests of binding theory. AB - This study reports effects of meaning and emotion (taboo vs. neutral words) on an illusory word (IW) phenomenon linked to orthographic repetition blindness (RB). Participants immediately recalled rapid serial visual presentation (RSVP) lists consisting of two critical words (C1 and C2) containing shared letters, followed by a word fragment: for example, lake (C1) brake (C2) ush (fragment). For neutral critical words, participants often recalled C1, but not C2 or the fragment, reporting instead a nonoccurring or illusory word: here, brush (a blend of C2 and the fragment). Forward RB (defined as reduced report of orthographically similar C2s) was more common for neutral than for taboo C2s, and taboo IWs were reported significantly more often than were neutral IWs. Moreover, when both C2 and the potential IW were taboo, a new phenomenon emerged: Participants reliably reported both the IW and the intact C2. These and other results supported a binding theory of the IW phenomenon and orthographic RB. PMID- 16365953 TI - John Raven. PMID- 16365954 TI - Standards of evidence: criteria for efficacy, effectiveness and dissemination. AB - Ever increasing demands for accountability, together with the proliferation of lists of evidence-based prevention programs and policies, led the Society for Prevention Research to charge a committee with establishing standards for identifying effective prevention programs and policies. Recognizing that interventions that are effective and ready for dissemination are a subset of effective programs and policies, and that effective programs and policies are a subset of efficacious interventions, SPR's Standards Committee developed overlapping sets of standards. We designed these Standards to assist practitioners, policy makers, and administrators to determine which interventions are efficacious, which are effective, and which are ready for dissemination. Under these Standards, an efficacious intervention will have been tested in at least two rigorous trials that (1) involved defined samples from defined populations, (2) used psychometrically sound measures and data collection procedures; (3) analyzed their data with rigorous statistical approaches; (4) showed consistent positive effects (without serious iatrogenic effects); and (5) reported at least one significant long-term follow-up. An effective intervention under these Standards will not only meet all standards for efficacious interventions, but also will have (1) manuals, appropriate training, and technical support available to allow third parties to adopt and implement the intervention; (2) been evaluated under real-world conditions in studies that included sound measurement of the level of implementation and engagement of the target audience (in both the intervention and control conditions); (3) indicated the practical importance of intervention outcome effects; and (4) clearly demonstrated to whom intervention findings can be generalized. An intervention recognized as ready for broad dissemination under these Standards will not only meet all standards for efficacious and effective interventions, but will also provide (1) evidence of the ability to "go to scale"; (2) clear cost information; and (3) monitoring and evaluation tools so that adopting agencies can monitor or evaluate how well the intervention works in their settings. Finally, the Standards Committee identified possible standards desirable for current and future areas of prevention science as the field develops. If successful, these Standards will inform efforts in the field to find prevention programs and policies that are of proven efficacy, effectiveness, or readiness for adoption and will guide prevention scientists as they seek to discover, research, and bring to the field new prevention programs and policies. PMID- 16365955 TI - Screening for cancer: testicular and prostate cancer. PMID- 16365956 TI - Saunas and your health. PMID- 16365957 TI - Grapefruit and medication: a cautionary note. PMID- 16365958 TI - On call. I had a hip replacement operation eight years ago and I've had a wonderful result. Now I'm ready to have my other hip done. Last time, I had blood thinning shots in the hospital, but this time my orthopedist also wants me to take a pill at home for a month. Is it really necessary? PMID- 16365959 TI - Canadian developments. Recommendations published concerning non-disclosure of HIV status. AB - The conclusions from an expert working group on persons who fail to disclose their HIV status were published in the 1 March 2005 edition of the Public Health Agency of Canada's Communicable Disease Report. The recommendations recognize that legal and ethical considerations must inform and guide both policy and practice as they relate to non-disclosure of HIV/AIDS. The expert working group favoured a graduated response model based on a public health approached to the issue rather than a criminal law approach. PMID- 16365960 TI - Canadian developments. Criminal charges laid in three new situations. PMID- 16365961 TI - Canadian developments. Buprenorphine now approved for the treatment of opiate addiction. PMID- 16365962 TI - Saskatchewan: Mandatory "bodily substances" testing legislation passed. PMID- 16365963 TI - First evaluation of Vancouver safe injection facility. PMID- 16365964 TI - Officials in two Canadian cities express support for safe injection sites. PMID- 16365965 TI - New Brunswick: sex education curriculum adds abstinence statement. PMID- 16365966 TI - Many HIV-positive people do not get enough to eat. PMID- 16365967 TI - ABC in Uganda: success or subterfuge? AB - Since the enactment of the US President's Emergency Plan for AIDS Relief (PEPFAR) in 2003, US officials have repeatedly pointed to Uganda as an example of the effectiveness of the "ABC" (Abstain, Be faithful, use Condoms) approach to HIV prevention. Uganda is one of few African countries to have experienced a decline in national HIV prevalence, from approximately 15 percent in the early 1990s to an estimated six percent today. While scholars continue to debate the reasons for this, the US government has attributed the decline to increased abstinence, faithfulness and condom use among Ugandans, and thus has made ABC the centerpiece of its global HIV prevention strategy. PMID- 16365968 TI - UN leadership and harm reduction: a rough road. AB - An unprecedented coalition of over 300 harm reduction, human rights and HIV/AIDS focused NGOs from around the world worked together in support of harm reduction in the lead-up to the annual session of the UN Commission on Narcotic Drugs (CND) in March 2005. PMID- 16365969 TI - United States: Funding restrictions threaten sex workers' rights. AB - Recent developments concerning the US government's restrictive policies on HIV/AIDS funding have drawn attention to how the government's mandatory "anti prostitution pledge" endangers the lives of sex workers and trafficking victims. PMID- 16365970 TI - Russian Federation: Going backwards on drug policy. AB - On 6 April 2005, the Russian government submitted a bill to the Duma (the national parliament) which, if passed, would turn back many of the recent reforms of Russia's drug policy. PMID- 16365971 TI - India: New patent law may restrict access to HIV/AIDS treatments. AB - On 4 April 2005, India's Patents (Amendment) Act, 2005 (the Act) received presidential assent. The Act, which represents a major shift in Indian patent policy, raises serious concerns that the supply of generic drugs to developing countries could decline. PMID- 16365972 TI - Taiwan: AIDS NGOs fight to keep human rights law. AB - Thirty-seven government deputies, all members of the ruling Democratic Progress Party (DPP), have recently proposed a bill to eliminate Article 6-1 of Taiwan's AIDS Prevention and Control Act (1997). PMID- 16365973 TI - Tenofovir trials raise ethical issues. AB - The development of safe, effective and accessible prevention methods has become one of the most urgent global public health needs. Whether the antiretroviral drug tenofovir (Viread), a nucleotide reverse transcriptase inhibitor, is appropriate for use as pre-exposure prophylaxis (PREP) for the prevention of HIV infection is currently being studied in a number of clinical trials. However, recent controversies over perceived defects in trial design and implementation, and inadequate consultation with the communities involved, threaten to prevent ongoing research unless closer collaboration between researchers and activists can be established. PMID- 16365975 TI - Holland: companies to offer life insurance to people living with HIV/AIDS. PMID- 16365974 TI - US: developments in the treatment of HIV-positive prisoners in two states. AB - Legal actions have been launched in Alabama and Mississippi to address living conditions and medical care of HIV-positive prisoners in state prisons. These were the only two states to allow complete segregation of HIV-positive prisoners in state prisons into the 1990s. The two cases highlight the ways in which the courts have been involved in supervising prison conditions in the United States. PMID- 16365976 TI - Nepal: Sexual minorities group facing possible ban. PMID- 16365977 TI - Australia: new free trade agreement with US targets drug prices. PMID- 16365978 TI - Scottish Prison Service to halt mandatory drug testing. PMID- 16365979 TI - HIV/AIDS in the courts--Canada. Federal Court overturns negative Pre-Removal Risk Assessment of HIV-positive failed refugee claimant. AB - An HIV-positive failed refugee claimant from Mexico has successfully challenged a Pre-Removal Risk Assessment (PRRA) decision by a Canadian immigration officer. PMID- 16365980 TI - HIV/AIDS in the courts--Canada. Argentinean couple living with HIV denied Convention refugee status. AB - On 25 January 2005, the Federal Court of Canada upheld an Immigration and Refugee Board (IRB) decision refusing to grant refugee status to an HIV-positive man and his same-sex partner. Pablo Sergio Gonzalez and his partner, Nelson Horacio De Robles, feared persecution by the government of Argentina because of Gonzalez's past political activity. PMID- 16365981 TI - HIV/AIDS in the courts--Canada. Landlord found to have discriminated based on HIV status. AB - The British Columbia Human Rights Tribunal (the Tribunal) has allowed a claim for discrimination against a Vancouver landlord who refused to rent an apartment to a couple, both of whom were living with HIV/AIDS. PMID- 16365982 TI - HIV/AIDS in the courts--Canada. Supreme Court finds settlement money attributable to past disability benefits taxable. AB - The Supreme Court of Canada recently held that the portion of a lump sum settlement from a lawsuit attributable to past disability insurance benefits should be included as taxable income. PMID- 16365983 TI - HIV/AIDS in the courts--Canada. Court affirms that severance exclusion for workers with disabilities violates Charter. AB - The Ontario Court of Appeal has upheld a lower court decision declaring a severance provision of the provincial Employment Standards Act (the Act) to be unconstitutional. Section 58(5)(c) of the Act, which denies severance pay to employees whose jobs are terminated due to illness or injury, was found to be of no force and effect by the Ontario Divisional Court in January 2004. On 4 May 2005, the Court of Appeal affirmed that the provision contravenes section 15(1) of the Canadian Charter of Rights and Freedoms. PMID- 16365984 TI - HIV/AIDS in the courts--Canada. Tribunal finds Quebec ministry's refusal to cover HIV medication "not a serious question". PMID- 16365985 TI - Australia: HIV-positive visa applicants successfully appeal refusals based on medical inadmissibility. AB - Two visa applicants have successfully appealed decisions to refuse their applications on the basis of medical inadmissibility, claiming that their HIV status does not present significant costs to the Australian health care system.[ PMID- 16365986 TI - UK: House of Lords upholds deportation order. AB - On 5 May 2005, the House of Lords rejected a 30-year old HIV-positive woman's appeal of her deportation order. The woman had argued that expelling her to Uganda, a country where access to HIV medication and medical care was uncertain, constituted a violation of guarantees against inhuman treatment in the European Convention (the Convention). The House of Lords ruled that deporting an HIV positive asylum seeker is not a violation of the Convention absent "exceptional circumstances." PMID- 16365987 TI - Sweden's compulsory confinement order declared a violation of liberty guarantee. AB - On 25 January 2005, the European Court of Human Rights unanimously held that the Swedish government violated an HIV-positive man's right to liberty when it placed him under a detention order for several years. PMID- 16365988 TI - UK: legal action on needle exchange programs in prisons dismissed. AB - In April 2005, a judicial review application against the UK Home Secretary for his failure to introduce needle exchange programs in prisons in England and Wales was dismissed by a judge at the Royal Courts of Justice (Administrative Court Division). PMID- 16365989 TI - Europe: Court declares admissible complaint about lack of medical assistance in Russian detention facility. AB - On 3 March 2005, the European Court of Human Rights found admissible an application by Viktor Vasilyevich Khudobin, a Russian national, who claimed that his arrest and detention by Russian authorities had violated several provisions of the European Convention on Human Rights (the Convention). PMID- 16365990 TI - South Africa: book publisher ordered to pay damages for disclosing women's HIV status. AB - The High Court of South Africa found that a book publisher had violated the constitutional rights of three HIV-positive women by publishing their names and HIV-positive status. In their claim, the women named as defendants the book's author, a prominent journalist, the book's subject, politician Patricia de Lille, and the publisher, New Africa Book Publishers. PMID- 16365991 TI - Criminal law and HIV transmission or exposure: four new cases. PMID- 16365992 TI - Australia: refusal to provide tattooing services found not discriminatory. PMID- 16365993 TI - Japan: High Court upholds health official's conviction on one charge, dismisses another. PMID- 16365994 TI - New Zealand: woman denied compensation for exposure to HIV. PMID- 16365995 TI - XI Mediterranean Congress of Rheumatology, Heraklion, Crete, 22-24 September 2005. Abstracts. PMID- 16365996 TI - Evaluation of environmental data for identification of Anopheles (Diptera: Culicidae) aquatic larval habitats in Kisumu and Malindi, Kenya. AB - This research evaluates the extent to which use of environmental data acquired from field and satellite surveys enhances predictions of urban mosquito counts. Mosquito larval habitats were sampled, and multispectral thermal imager (MTI) satellite data in the visible spectrum at 5-m resolution were acquired for Kisumu and Malindi, Kenya, during February and March 2001. All entomological parameters were collected from January to May 2001, June to August 2002, and June to August 2003. In a Poisson model specification, for Anopheles funestus Giles, shade was the best predictor, whereas substrate was the best predictor for Anopheles gambiae, and vegetation for Anopheles arabensis Patton. The top predictors found with a logistic regression model specification were habitat size for An. gambiae Giles, pollution for An. arabensis, and shade for An. funestus. All other coefficients for canopy, debris, habitat nature, permanency, emergent plants, algae, pollution, turbidity, organic materials, all MTI waveband frequencies, distance to the nearest house, distance to the nearest domestic animal, and all land use land cover changes were nonsignificant. MTI data at 5-m spatial resolution do not have an additional predictive value for mosquito counts when adjusted for field-based ecological data. PMID- 16365997 TI - No evidence for bacteriophage WO orf7 correlation with Wolbachia-induced cytoplasmic incompatibility in the Culex pipiens complex (Culicidae: Diptera). AB - Gene flow between populations of Culex pipiens L. is relevant to observed differences in disease transmission, insecticide resistance, behavior, and physiology. Intracellular Wolbachia bacteria have been hypothesized to affect gene flow in insects. Specifically, Wolbachia cause a form of embryonic mortality known as cytoplasmic incompatibility (CI) in crosses between individuals with different Wolbachia types. Incompatibility in Culex is exceptional in that it represents the most complex CI pattern known, and yet Culex populations are not infected with divergent Wolbachia strains. This has led to the hypothesis that extrachromosomal factors such as phages or mobile genetic elements may be involved in determining CI phenotype. Recent molecular characterization of Culex laboratory strains has identified variation in the orf7 locus of the Wolbachia associated bacteriophage WO. Here, crosses between eight Culex strains differing in their orf7 type were conducted to examine for the hypothesized involvement of bacteriophage WO in determining CI in Culex. Although crossing results show examples of compatibility, partial compatibility, and incompatibility, the results fail to show a correlation between the CI phenotypes and orf7 type. Specific examples include high egg hatch resulting in crosses between Culex strains that differ significantly in their orf7 type and low egg hatch resulting in crosses between Culex strains with similar orf7 types. Thus, the phage orf7 locus alone cannot predict CI type in the Culex strains examined in this study. However, rejection of the hypothesized role of WO phage in Culex CI will require the characterization of additional phage loci. PMID- 16365998 TI - Intragenomic heterogeneity of internal transcribed spacer rDNA in neotropical malaria vector Anopheles aquasalis (Diptera: Culicidae). AB - Intragenomic heterogeneity of the internal transcribed spacer (ITS) array was investigated in Anopheles aquasalis Curry mosquitoes from two geographic locations in each of Brazil and Venezuela, and one in Suriname. Polymerase chain reaction-amplified copies of the ITS were cloned and sequenced. The length of the entire array ranged from 782 to 990 bp, with most variation due to microsatellite insertions in ITS1. We detected 40 different ITSL sequences and 15 different ITS2 sequences of the 71 to 72 clones examined. The sequence divergence within localities ranged from 0.002 to 0.043 for ITS1 and from 0 to 0.006 for ITS2. Point mutations were common to both spacer regions, but dinucleotide microsatellite repeats were restricted to ITS1. Sequences from neither ITS1 nor ITS2 had a diagnostic distribution or were informative in distinguishing these populations, providing additional support for the status of An. aquasalis as a single species. PMID- 16365999 TI - Susceptibility of Psorophora confinnis (Diptera: Culicidae) to infection with epizootic (subtype IC) and enzootic (subtype ID) Venezuelan Equine encephalitis viruses. AB - To test the hypothesis that adaptation to epizootic mosquito vectors mediates the emergence of Venezuelan equine encephalitis virus (family Togaviridae, genus Alphavirus, VEEV) from enzootic progenitors, the susceptibility of the epizootic vector Psorophora confinnis (Lynch-Arribalzaga) to epizootic versus enzootic strains was evaluated. Artificial bloodmeals containing subtype IC strains isolated during the 1962-1964, 1992-1993, and 1995 Venezuelan/Colombian epizootics and closely related Venezuelan enzootic subtype ID strains were used to compare mosquito infectivity and transmission potential. Strains from the smaller 1992-1993 epizootic showed lower or equal infectivity and replication compared with enzootic viruses and to strains isolated during the larger 1962 1964 and 1995 epizootics. These experiments failed to provide evidence that Ps. confinnis selects for epizootic VEEV viruses with higher infectivity, as has been shown for Aedes (Ochlerotatus) taeniorhynchus (Wiedemann). Nonetheless, its high susceptibility, abundance in enzootic and epizootic regions, and feeding behavior suggest that Ps. confinnis is an important bridge vector for both enzootic and epizootic VEEV. PMID- 16366000 TI - Capillary tube feeding system for studying tick-pathogen interactions of Dermacentor variabilis (Acari: Ixodidae) and Anaplasma marginale (Rickettsiales: Anaplasmataceae). AB - A capillary tube feeding (CTF) system was adapted for studying the interaction between Dermacentor variabilis (Say) and the rickettsial cattle pathogen Anaplasma marginale Theiler. A. marginale undergoes a complex developmental cycle in ticks that begins in midguts and ends by transmission from salivary glands. In this CTF system, male D. variabilis were fed A. marginale-infected blood or cultured tick cells. Ticks that fed on highly rickettsemic calves developed midgut and salivary gland infections as detected by PCR, whereas ticks that were fed from capillary tubes on the same blood developed only midgut infections. An unexpected result of capillary tube feeding was that antibodies against the A. marginale adhesin, major surface protein la, enhanced midgut infections and caused cell culture-derived A. marginale to infect midguts. Another unexpected result was the infection of the midguts of the nonvector tick Amblyomma americanum (L.), after capillary tube feeding on infected bovine blood. The gut cell response of ticks to A. marginale, as determined from SDS-polyacrylamide gel electrophoresis protein profiles, did not differ when ticks were fed infected or uninfected cells from capillary tubes. Selected protein bands, as identified by tryptic digestion-mass spectrometry, contained mostly proteins of bovine origin, including bovine albumin, undigested alpha- and beta-chain hemoglobin and hemoglobin fragments. Although infection of ticks by A. marginale CTF system was not the same as infection by feeding on cattle, the results obtained demonstrated the potential use of this system for identifying aspects of pathogen-vector interactions that are not readily recognized in naturally feeding ticks. PMID- 16366001 TI - Isolation of viruses from mosquitoes (Diptera: Culicidae) collected in the Amazon Basin region of Peru. AB - As part of a comprehensive study on the ecology of arthropod-borne viruses in the Amazon Basin region of Peru, we assayed 539,694 mosquitoes captured in Loreto Department, Peru, for arboviruses. Mosquitoes were captured either by dry ice baited miniature light traps or with aspirators while mosquitoes were landing on human collectors, identified to species, and later tested on Vero cells for virus. In total, 164 virus isolations were made and included members of the Alphavirus (eastern equine encephalomyelitis, Trocara, Una, Venezuelan equine encephalomyelitis, and western equine encephalomyelitis viruses), Flavivirus (Ilheus and St. Louis encephalitis), and Orthobunyavirus (Caraparu, Itaqui, Mirim, Murutucu, and Wyeomyia viruses) genera. In addition, several viruses distinct from the above-mentioned genera were identified to the serogroup level. Eastern equine encephalomyelitis virus was associated primarily with Culex pedroi Sirivanakarn & Belkin, whereas Venezuelan equine encephalomyelitis virus was associated primarily with Culex gnomatos Sallum, Huchings & Ferreira. Most isolations of Ilheus virus were made from Psorophora ferox (Von Humboldt). Although species of the Culex subgenus Melanoconion accounted for only 45% of the mosquitoes collected, 85% of the virus isolations were made from this subgenus. Knowledge of the viruses that are being transmitted in the Amazon Basin region of Peru will enable the development of more effective diagnostic assays, more efficient and rapid diagnoses of clinical illnesses caused by these pathogens, risk analysis for military/civilian operations, and development of potential disease control measures. PMID- 16366002 TI - Shiga toxin 2 and lipopolysaccharide cause monocytic THP-1 cells to release factors which activate platelet function. AB - Platelet and monocyte activation may contribute to hemolytic anemia, thrombocytopenia and renal failure associated with the hemolytic uremic syndrome (HUS) caused by Escherichia coli O157:H7. Since Shiga toxins (Stxs) and lipopolysaccharide (LPS) from this bacterium are implicated in the pathogenesis of HUS, we examined whether stimulation of THP-1 human monocytic cells by Shiga toxin 2 (Stx2) and LPS can lead to the activation of platelet function. We now show that Stx2 causedTHP-1 cells to release the chemokines IL-8, MDC, and RANTES and that the presence of LPS further stimulated this release. IL-8 was produced in greatest amount and was an effective co-agonist for inducing platelet aggregation. Primary human monocytes also released large amounts of IL-8 in response to LPS and Stx2. Factors released byTHP-1 cells exposed to Stx2 and LPS activated platelet function as evidenced by increased aggregation, serotonin secretion, P-selectin exposure and by the formation of stable platelet-monocyte aggregates. Our data therefore show that monocytes exposed to E.coli-derived Stx2 and LPS release factors which activate platelet function. PMID- 16366003 TI - Modulation of the intestinal response to ionizing radiation by anticoagulant and non-anticoagulant heparins. AB - Endothelial dysfunction is involved in radiation responses in many normal tissues, including intestine. Endothelium-directed interventions ameliorate intestinal radiation injury (radiation enteropathy) in animal models, and anecdotal reports also suggest a beneficial effect of heparin. This study assessed low molecular weight heparin as an intestinal radiation response modifier. Rats underwent localized small bowel irradiation. Groups of rats were treated with saline, nadroparin (3 mg/kg/d), or a non-anticoagulant heparin (SR80258, 3 mg/kg/d), from 3 days before to 2 weeks after irradiation. The intestinal radiation response was assessed 2 weeks and 6 weeks after irradiation using quantitative histology; morphometry, and cellular and molecular end-points. Compared to vehicle-treated controls, nadroparin significantly exacerbated structural radiation injury, neutrophil infiltration, and TGFbeta and collagen I immunoreactivity levels 2 weeks after irradiation. SR80258 was associated with increased TGFbeta levels, but the other parameters did not reach statistical significance. At 6 weeks, structural, cellular, and molecular injury was similar in the three experimental groups. Heparin, in contrast to antiplatelet agents and direct thrombin inhibitors, does not ameliorate, but exacerbates acute intestinal radiation toxicity. These data underscore the importance of heparin as an inhibitor of physiological anti-inflammatory mechanisms during tissue injury, as well as the non-anticoagulant effects of heparin. Moreover, these data may have implications for the use of heparin during radiation therapy. PMID- 16366004 TI - Occupational injuries among construction workers treated in a major metropolitan emergency department in the United States. AB - OBJECTIVES: The aim of this study was to profile construction workers' injuries for more information about the causes of nonfatal construction worker injuries and identify injury trends for further investigations and prevention programs. METHODS: An injury-tracking program for emergency departments was established in 1990 to gather the data needed for the study. Profiles were obtained for 2916 construction workers' injuries that were identified on hospital registration forms at the George Washington University Emergency Department in Washington, DC, from November 1990 through October 1997. Laborers and construction workers who did not specify a trade were combined, and together they made up the largest group--29% of the injured workers. RESULTS: The leading cause of injury was contact with cutting or piercing objects-most often pieces of metal, razors, knives, power tools, and nails. Workers striking against objects or being struck by objects (including falling objects) accounted for the second-largest group of injuries, and the third leading injury circumstance was falling--either from a height or on the same level. Detailed injury statistics are presented by trade, showing patterns of injury that reflect tasks of these trades and which injuries predominated in each trade. Although many previous reports have described construction workers' injuries, very few have provided detailed data by trade. CONCLUSIONS: The details presented in this analysis allow for a better understanding of the injury circumstances and provide a starting point for injury prevention programs. PMID- 16366005 TI - [Adult-onset hypophosphatemic osteomalacia]. PMID- 16366006 TI - Effect of source spectral shape on task-based assessment of detection and resolution in optical coherence tomography. AB - We demonstrate the effect of the spectral shape of broadband light sources in a task-based approach for assessment of signal detection and resolution in optical coherence tomography. We define two binary tasks: The signal is either present or absent and the signal can be either resolved or not. In a transparent sample bounded by two uniform interfaces we study the minimum detectable change in the index of refraction as well as the minimum resolvable distance between the layers in correlation with the source spectral shape and power. Results show that the area under the receiver operating curve (AUC) for a signal-detection task is not affected by the shape of the spectrum but solely by its optical power, whereas spectral shaping has an effect, which we quantify, on the AUC for the resolution task. Moreover, the AUC is demonstrated in relation to the concept of system sensitivity for a signal-detection task. PMID- 16366007 TI - Motivational interviewing and clinical psychiatry. AB - OBJECTIVES: Our objectives were as follows: 1) to survey the literature on motivational interviewing (MI), "a client-centered yet directive method for enhancing intrinsic motivation to change by exploring and resolving client ambivalence" and a well-established method of brief intervention, especially in the field of addictions treatment; 2) to review hypothesis about its mode of action; and 3) to discuss its possible impact on clinical psychiatry, in particular, on teaching communication skills. METHOD: Literature reviews and metaanalyses of numerous clinical trials of MI for addictions treatment have already been published and are briefly summarised. So far, no literature survey exists for MI applied to psychiatric patients. This review is limited to a synthesis of the articles relevant to psychiatry and to comments based on our team's experiences with MI. RESULTS: There is no evidence that MI achieves better results than our other established techniques for treating addictions; it may simply work faster. The explanation for the method's rapid effectiveness remains speculative. Outcomes concerning the application of MI to psychiatric patients, although preliminary, are promising. Methods of assessing the integrity of MI treatment are more developed than in most psychotherapies, which permits the learning progress of trainees to be measured. CONCLUSIONS: MI offers a complement to usual psychiatric procedures. It may be worthwhile to teach it, not only for addictions but also for other broad treatment issues, such as enhancing patients' medication compliance and professionals' communication skills. Questions remain concerning MI's feasibility in psychiatry settings. PMID- 16366008 TI - Breast cancer among airline cabin attendants. PMID- 16366009 TI - [Celiac disease]. PMID- 16366010 TI - For the record... PMID- 16366011 TI - Proceedings of the 16th Novo Nordisk Symposium on Growth Hormone and Endocrinology, Vienna, Austria, April 1-2, 2005. PMID- 16366013 TI - Research in Medical Education. Proceedings of the 44th Annual Conference, November 6-9, 2005, Washington, DC, USA. PMID- 16366012 TI - Proceedings and abstracts of the VII Symposium Progress in Clinical and Forensic Toxicology, 2-4 June 2005, Krakow, Poland. PMID- 16366014 TI - [Block formation in Citellophilus Tesquorum altaicus Ioff, 1936, fleas in relation to the sex of an insect]. AB - Males and females of C. Tesquorum altaicus, the main vector of plague in its natural focus, were experimentally studied in different seasons (spring, summer) for their capacity of block formation. The experiments used 2 groups of fleas: the insects that have been infected in last autumn and hibernated and <> insects, a that year's breeding. The investigations demonstrated that the hibernated fleas had insignificant differences (P>0.05) in the rates of block formation by sex in spring and early summer. Among the <> fleas, a that year's breeding, the males with a gizzard block considerably exceeded the females in number (P<0.05) in July-August (a period of an intensified epizootic process in the focus). By seasons, the rates of block formation were 7.0 to 21.7% in males and 2.0 to 14.0% in females. PMID- 16366015 TI - [Organization and implementation of prophylactic measures in the epidemiological foci of tick-borne encephalitis in the Volyn region]. PMID- 16366016 TI - [Possible influences of climatic changes on the spread and intensity of foci of some human helminthiases]. AB - Climatic changes on the Earth and in some of its regions have caused and will cause alterations in the natural and social factors which may influence the circulation of causative agents of helminthiases that develop both on land and in water with the participation of hydrobiont. These include changes in the size of the first and second intermediate hosts, in the hydrological conditions of water reservoirs, in the amount of precipitation, soil moisture, heat, etc. Therefore, the expected global warming may alter the areas of some helminths, the intensity of foci, etc. PMID- 16366017 TI - Medical abortion: expanding access to safe abortion and saving women's lives. PMID- 16366018 TI - Medical abortion: a fact sheet. PMID- 16366020 TI - National Hospital Ambulatory Medical Care Survey: 2003 outpatient department summary. AB - OBJECTIVES: This report describes ambulatory care visits to hospital outpatient departments (OPDs) in the United States. Statistics are shown on selected hospital, clinic, patient, and visit characteristics, as well as selected trends in OPD visits since 1993. The report highlights variation in use across the major types of OPD clinics surveyed. METHODS: The data shown in this report were collected from the 2003 National Hospital Ambulatory Medical Care Survey (NHAMCS). NHAMCS is a national probability sample survey of visits to emergency and outpatient departments of non-Federal, short stay, and general hospitals in the United States. Sample data are weighted to produce annual national estimates. RESULTS: During 2003, an estimated 94.6 million visits were made to hospital OPDs in the United States, about 33.1 visits per 100 persons. This rate represents a 35-percent increase since 1993, although rates have been stable since 1999. Infants under 12 months of age had a visit rate of 88.7 visits per 100 persons, a rate that increased by 23 percent since 1993. Increasing trends in OPD visit rates were found for persons 50-64 years of age (up by 30 percent), 13-21 years of age (up by 32 percent), 22-49 years of age (up by 34 percent), and 1-12 years of age (up by 71 percent). Females had higher OPD visit rates than males (39.6 compared with 26.4 visits per 100 persons), and black or African American persons had higher OPD visit rates than white persons (59.7 compared with 29.9 visits per 100 persons). Medicaid and State Children's Health Insurance Program patients used OPDs for preventive care services more frequently than private pay patients. The preventive care visit rate by Hispanic and Latino patients was twice the rate by non-Hispanic patients. Diphtheria, tetanus, and acellular pertussis (DTaP) was the most frequently provided vaccine to children under age 18 years. Between 1993 94 and 2003, the proportion of visits involving only mid-level providers increased from 5.9 to 12.6 percent of visits. PMID- 16366021 TI - [Opportunities for nurses]. PMID- 16366022 TI - [Nobody should lose out now]. PMID- 16366023 TI - [New tariffs: waiting--but keeping the eyes open]. PMID- 16366024 TI - [Computer-generated communication assistance]. PMID- 16366025 TI - [Nursing involved in structural changes]. PMID- 16366026 TI - [The Bologna process in nursing education]. PMID- 16366027 TI - [Patients should not suffer pain]. PMID- 16366028 TI - [New dimensions of professionalism in Europe]. PMID- 16366029 TI - [Creating modules in the area of development]. PMID- 16366030 TI - [Nursing initiative for education for girls in Africa]. PMID- 16366031 TI - [Nursing 2006: Congress for rights, management and practice]. PMID- 16366032 TI - [Equal treatment]. PMID- 16366033 TI - [Pathobiochemical importance of phospholipases for the release of mast cell mediators]. AB - We investigated the influence of polyunsaturated fatty acids on the activity of the cytosolic phospholipase A2 (cPLA2) in the canine mastocytoma cell line C2 as a model for canine atopic dermatitis (CAD). Cells were cultured in a basic medium or in media supplemented with different fatty acids (14 microM) for eight days. The supplemented fatty acids were linoleic acid (18:2n6), alpha-linolenic acid (18:3n3), gamma-linolenic acid (18:3n6) and docosahexaenoic acid (22:6n3). We measured enriched concentrations of the added fatty acid, their delta6 desaturated and elongated products. However, delta5-desaturated products were not increased. Culturing of C2 in 18:3n3 supplemented medium reduced the cPLA2 activity. Furthermore in these cells and in C2 cultured in 22:6n3 supplemented medium decreased the cPLA2 activity after stimulation. The reduced cPLA2 activity by the changed fatty acid pattern of C2 cultured in 18:3n3 or 22:6n3 possibly explain the beneficial effects of these fatty acids in CAD because increased cPLA2 activity is accompanied by enhanced release of proinflammatory type 2 prostaglandins and type 4 leukotrienes. PMID- 16366034 TI - [Contents of the glucose transporters GLUT1 and GLUT4 in oxidative and glycolytic muscles of goat kids and adult goats]. AB - The objective of this study was to elucidate, whether the impaired insulin sensitivity with regard to glucose utilisation in ruminating goats compared with suckling goat kids may be due to a reduced expression of the glucose transporters GLUT1 and GLUT4 in skeletal muscle. Muscle samples were removed from red, oxidative muscles (M. masseter, diaphragm) and white, glycolytic muscles (M. longissimus dorsi, and M. semitendinosus) of five goat kids fed with milk exchanger and nine adult goats of different stages of life. Samples were analysed for their GLUT1 and GLUT4 contents. The muscles were characterised metabolically by measuring the activities of isocitrate dehydrogenase (ICDH) and of lactate dehydrogenase. In all four analysed muscles the GLUT4 contents of adult goats were significantly (p < 0.05) lower than in goat kids. Significant differences concerning the GLUT4 contents in skeletal muscles were not detected in adult goats of different stages of life. The GLUT1 contents differed to a lower extend between goat kids and adult goats. The results of this investigation indicate that the impaired insulin sensitivity of adult compared with suckling ruminants is accompanied by or leads to a decreased GLUT4 expression. PMID- 16366035 TI - [Large intestinal epithelium and short-chain fatty acids. Expected, unexpected, unexplained findings]. AB - Short-chain fatty acids (SCFA) are present in hindgut contents in high concentrations. SCFA are generated and also absorbed rapidly in the large intestine. Absorption results from diffusion of the undissociated and lipid soluble form or in exchange for bicarbonate. The controversial concepts concerning the extent of diffusion or the exchange for bicarbonate are partly due to contradictory findings and unequal mechanisms in different species and in different segments of the large intestine as well as in the different methods used. An almost neutral pH microclimate at the luminal surface is of importance for absorption of SCFA. The apical membranes of the epithelial cells in caecum and in proximal colon of guinea pigs are an substantial barrier for the diffusion of SCFA. After withdrawal of butyrate for only one hour from the perfusion or incubation solutions a massive apoptosis had been observed during the in situ perfusion of segments of guinea pig large intestine and also in in vitro studies with isolated epithelia in Ussing-chambers. In vitro apoptotic bodies and cells are emitted at the epithelial surface. However, in vivo induced by butyrate deprivation resident macrophages were tightly clustered below the surface epithelium. Aided by cytoplasmatic projections these macrophages phagocytose and transport apoptotic material from the epithelial intercellular spaces into their bodies. Obviously macrophages can be overloaded by this massive apoptosis, and some of the undigested apoptotic fragments are emitted into the lamina propria. Formation of a colitis ulcerosa may originate from these released undigested apoptotic bodies. Furthermore hints indicate aetiological interrelations between deprivation of butyrate and colon cancer. Butyrate-paradox denotes the contrarian effects on apoptosis and cell proliferation after addition or deprivation of butyrate in cultures of large intestinal tumour-cell-lines in comparison with the healthy, intact epithelium. PMID- 16366036 TI - [High intestinal transport activity for nucleosides in cattle: a synopsis]. AB - In ruminants large amounts of nucleic acids associated with the microbial cell mass leaving the fore-stomach system via the abomasum enter the small intestine. In dairy cows this amounts to 100-200 g microbial nucleic acids per day. These nucleic acids are very efficiently digested in the small intestine whereby nucleosides were found to be the main degradation products. Therefore, this review centers mainly on the mechanisms of intestinal absorption of nucleosides and their role in nucleic acid digestion. Furthermore, metabolism of nucleosides in intestinal epithelial cells and its bearing for nucleoside absorption and salvage of nucleosides for nucleic acid synthesis in various tissues is considered. According to own studies using isolated intestinal brush-border membrane (BBM) vesicles (BBMV) from dairy cows purine and pyimidine nucleosides are transported actively by two separate Na+ co-transport systems (N1 and N2) across the bovine BBM, whereby transport activity in the small intestine decreases from proximal to distal. Guanosine and inosine appeared to be transported exclusively by N1 while thymidine and cytidine appeared to be transported exclusively by N2. Uridine and adenosine had an affinity to both transporters. In comparison to findings in man and rabbit, transport capacity (Vmax) of N1 and N2 in the BBM of cows was more than 10-fold higher. Similar findings were obtained in BBMV isolated from the small intestine of veal calves with rudimentary forestomach development in regard to nucleoside transport. Therefore, the high intestinal transport activity for nucleosides seems to be a genetically fixed property in the bovine, which is not related to a functional fore-stomach system. PMID- 16366038 TI - [The morphology of domestic pigs in middle Europe during the 16th to 18th century]. AB - The external of swine in Mideurope during the 16th to 18th century will be described based on a few publications and various pictures (120). Long legged, slender, bristly pigs with long snouts and prick ears dominated during the 16th century, sometimes, however, compact animals with shorter snouts and larger prick or lope ears have been depicted (Tab. 1). During the 17th century no correct descriptions of the pigs are avaiable. The pictures show similar types as in the century before (tab. 1). In pig herds the heterogenity especially of heads and ears in pigs kept in this time is demonstrated. During the 18th century agriculturists postulate long pigs as well as compact types. On the (few) pictures in this century more compact animals with short snouts and long ears have been presented, partly from England probably with the influence of imported asiatic pigs. From the 2 dominating types of land races in Mideurope at the end of the 18th century (small, prick eared or large with lope ears) the former may be derived from the medieval mideuropean pig, while the second one may arise from imports from mediterranean countries and crossings. PMID- 16366037 TI - [Utilization of glucose and long-chain fatty acids in lactating dairy cows fed a fat-enriched diet]. AB - The fate of carbon from long-chain fatty acids and glucose in dairy cows which were fed with protected fat was studied using stable isotope technique. The experiment was carried out on two groups of dairy cows (n=16 in each group) during the first 15 weeks of the lactation period. The cows were fed isoenergetic and isoproteinogenous diets based on corn silage. About 1.8 kg of tapioca starch in the diet of the starch group was substituted by about 0.7 kg of rumen protected fat (Ca salts of palm oil and soybean oil) in the diet of the fat group. The carbon atoms of dietary fat were naturally depleted in 13C as compared to carbon atoms of starch. Daily milk performance and lactose output were significantly (P < 0.05) higher among the cows fed with fat diet. In comparison to the starch group, the enrichment of milk fat with 13C was significantly lower, while that of breath CO2 was significantly higher in the fat group (P < 0.05). This means the fatty acids were incorporated into milk fat in preference to metabolic oxidation. Further studies showed that blood glucose is oxidized to a lower extent and is used for the synthesis of lactose to a higher proportion if the cows were fed with the fat diet. The glucose entry rate into the body glucose pool was not different between the diets. In conclusion, the dietary fatty acids perform a glucose sparing effect and improve the glucose supply for the mammary gland. PMID- 16366039 TI - [Pharmaco-toxicological and clinical studies with colocynth pulp extracts (Extr. colocynthidis fructus)]. AB - Colocynth pulp extract is a long-serving laxative. Contesting the official characterizations "drastic irritant action, no longer defensible" by suitable pharmacotoxicologic studies, extracts of the drug with increasing concentrations of the effective constituant "Cucurbitacins" were prepared in order to define efficacy ranges lethal to rats and mice. The extract Koloquinthentrockenextrakt Alpha with the highest content of Cucurbitacins (23,2 % delta 232,64 mg/g) permitted the definition of the LD50 for female (tentative because of death inhibition under maximal doses) and male rats; the mean LD50 = 281,8 and 525,6 mg/kg extract, respectively, equivalent to 66 and 122 mg/kg Cucurbitacins. This corresponds to 660- to 1220-fold therapeutic doses. Repeated administrations of 10- and 50-fold therapeutic doses to rats for 30 days produced no negative effects. The symptoms of rodent poisoning are described in detail. Pharmacologic doses were not toxic on rat liver slices, did not influence breathing and circulation parameters in guinea pigs (under the maximal dose of 41,6 mg/kg Cucurbitacins, 3/10 animals died of breathing failure) nor the behaviour of mice, nor were they mutagen (Ames test). Colocynth pulp extract weakly inhibited the growth of MDA-MB435 mamma carcinoma cells, but had no influence on the growth of B16 mouse melanoma cells. P388 mouse leukemia cells and L 929 mouse fibroblasts were not significantly influenced. High doses of Colocynth pulp extract inhibited diuresis and electrolyte excretion in rats. The Cucurbitacins E and I were rapidly metabolized in S9-supernatants of rat livers. A dried ethanolic Salvia fruit extract alleviated the toxicity of lethal doses of Colocynth pulp extract when administered simultaneously. A field study with 200 patients and a phase I study with 60 volunteers were conducted in Germany with Colocynth pulp extract from April to October 1998, andfrom December 2002 to March 2003, respectively. Data on the tolerance of the highest allowed dose and of a half-maximal dose administered to volunteers for 14 days in comparison to placebo, as well as data on the efficacy of a treatment course of 3 days of patients with obstipation, were to be gained. Clinical laboratory investigations of volunteers gave no indication of pathological changes even under the highest dose. In patients with obstipation and associated complaints, the administration for 3 days at maximum led to an increased frequency of bowel movements. At the same time, the discomforts accompanying obstipation were significantly relieved. Patients with obstipation defined tolerance as good. Volunteers, on the other hand, judged the tolerance of the drug significantly inferior ("good" - "average") to that of placebo ("very good" - "good"). The low risk potential of Colocynth pulp extract documented in pharmaco-toxicological studies was confirmed during administration to humans. PMID- 16366040 TI - Pharmacokinetics and comparative bioavailability of two vinpocetine tablet formulations in healthy volunteers by using the metabolite apovincaminic acid as pharmacokinetic parameter. AB - The objective of this study was to evaluate the pharmacokinetics of apovincaminic acid, the main metabolite of vinpocetine ((3alpha, 16alpha) -eburnamenine-14 carboxylic acid ethyl ester, CAS 42971-09-5), and to assess the average bioequivalence of two immediate release formulations of 10 mg vinpocetine tablets in 24 healthy male volunteers. The relative bioavailability of the test (generic) product (Vimpocetina) with respect to the reference product was determined in a single dose, randomized, crossover study. A simple, rapid specific and reliable high performance liquid chromatographic method coupled with mass spectrometry detection has been developed and validated for vinpocetine and apovincaminic acid. However, only the concentrations of the metabolite could be used for bioequivalence determinations because the concentrations of the parent drug were too low to be accurately measured in the biological matrix. The compartmental analysis of the metabolite's appearance-disappearance in blood shows similarity with first-order kinetics of a drug extravascularly administered. The apparent pharmacokinetic constants were determined. The mean values for the Cmax were 49.5 (+/- 16) ng/ml for test and 51.4 (+/- 14) ng/ml for the reference product. The mean values for the AUC0-infinity were 95 (+/- 29) ng/ml x h for test and 96.9 (+/- 26) ng/ml x h for reference, respectively. The 90 % confidence intervals for test/reference mean ratios of the plasma pharmacokinetic variables Cmax and AUC0 infinity lie between 0.83-1.08 and 0.88-1.08, respectively, which is within the conventional bioequivalence range of 80-125 % (Schuirman test). The difference between Tmax of the test and reference products was statistically non-significant (Friedman test). The test product is therefore bioequivalent to the reference product with respect to the rate and extent of apovincaminic acid pharmacokinetics. PMID- 16366041 TI - Efficacy and tolerability of a fixed combination of thyme and primrose root in patients with acute bronchitis. A double-blind, randomized, placebo-controlled clinical trial. AB - In a double-blind, randomized, placebo-controlled, multicenter, prospective study, the clinical efficacy and tolerability of a fixed combination of thyme fluid extract and primose root tincture (Bronchicum Tropfen) was investigated at a dosage of 30 drops (1 ml), taken orally five times daily. 150 outpatients (97 women, 53 men) suffering from acute, not previously treated bronchitis, lasting for less than 48 h, were randomized and treated with either verum (75 patients: 45 women, 30 men) or placebo (75 patients: 52 women, 23 men) over a time period of 7-9 days. 17 patients were excluded from the per-protocol (PP) collective because of either withdrawal from the trial (n = 2) or violations regarding examination time points and/or intake of the study medication (n = 15). The primary outcome criterion for efficacy assessment was the decrease of the Bronchitis Severity Score (BSS) at the end of the study compared to baseline. In the verum group, the BSS decreased from 12.0 +/- 4.4 points at baseline to 1.0 +/ 2.1 at study end compared to a decrease from 11.7 +/- 4.3 points at baseline to 6.5 +/- 4.8 at study end in the placebo group. The inter-group difference of 5.8 points was highly significant (p < or = 10(-3)) in favour of the verum medication [Intention-to-treat (ITT) analysis]. At the end of the study, significantly more patients were symptom free in the verum group (58.7%) than in the placebo group (5.3%) as compared by the ITT analysis (secondary outcome criterion). For both parameters, the PP analysis showed comparable results. The results for the concomitant variables of efficacy support the high superiority of the verum medication compared to placebo. The therapeutic effect was more pronounced the stronger the severity of the acute bronchitis was (as proofed by a stratified evaluation based on severity grade). The tolerability was very good in both groups; neither serious adverse events nor clinically relevant findings in the safety parameters were observed. A total of 7 adverse events occurred, 2 in the verum group and 5 in the placebo group. One of the two adverse events in the verum group was considered to be possibly related to the intake of the study medication. Neither serious nor unknown adverse drug reactions were observed. Two drop-outs occurred during the study related to adverse events, both in the placebo group. In the global safety assessment, the tolerability of both medications was rated as "good" or "very good" by more than 90% of the patients and physicians. In the present study, the fixed combination of thyme fluid extract and primrose root tincture was well tolerated and resulted in a clinically relevant and more pronounced decrease of the bronchitis symptoms (primary outcome criterion) and in shortening the duration of acute bronchitis (secondary outcome criterion) when compared to placebo. PMID- 16366042 TI - Mechanisms involved in the anti-inflammatory effect of a standardized willow bark extract. AB - A standardized willow bark extract (STW 33-I) has been examined to clarify its possible mechanism of action as an anti-inflammatory agent. Various facets have been investigated in two inflammation models: the 6-day air pouch model in rats, representing the acute state and the adjuvant induced arthritis representing the chronic one. Parameters included leukocytic infiltration, levels of cytokines and prostanoids in blood, and effects on cyclo-oxygenase (COX)-1 and/or COX-2 enzymes as well as effects involving free radical production. The effect of the extract was compared at two dose levels with comparable anti-inflammatory doses of acetylsalicylic acid (CAS 50-78-2, ASA) as a non-selective COX inhibitor, and celecoxib (CAS 169590-42-5) as a selective COX-2 inhibitor. On a mg/kg basis, the extract was at least as effective as ASA in reducing inflammatory exudates and in inhibiting leukocytic infiltration as well as in preventing the rise in cytokines, and was more effective than ASA in suppressing leukotrienes, but equally effective in suppressing prostaglandins. On COX-2, STW 33-I was more effective than ASA. The present findings show that STW 33-I significantly raises GSH (reduced glutathione) levels, an effect which helps to limit lipid peroxidation. The extract was more potent than either ASA or celecoxib. Higher doses of the extract also reduced malondialdehyde levels and raised shows definite superiority to either ASA or celecoxib in protecting the body against oxidative stress. It is therefore evident that STW 33-I is at least as active as ASA on all the parameters of inflammatory mediators measured, when both are given on a similar mg/kg dose. Considering, however, that the extract contains only 24% salicin (molecular weight 286.2), while ASA has a molecular weight of 180.3, it follows that on a molar basis of salicin vs salicylate, the extract contains less than a sixth of the amount of salicin as the amount of salicylate in ASA. Thus it appears that STW 33-I with its lower "salicin" content than an equivalent dose of ASA, is at least as active as ASA on the measured parameters, a fact that leads one to speculate that other constituents of the extract contribute to its overall activity. The presence of polyphenols in STW 33-I probably plays a significant role in enhancing its free radical scavenging properties. The fact that STW 33-I was superior to ASA in this respect would suggest that the extract may have a better anti-inflammatory effect than ASA on a weight to weight basis, with possibly less side effects. PMID- 16366043 TI - Bisphosphonates, hypercalcemia of malignancy, and osteonecrosis of the jaw. PMID- 16366044 TI - Impression success using the dual arch technique. AB - Taking an accurate impression and pouring a model that duplicates the clinical situation is the goal of every restorative dentist. The chance of having a restoration that fits the tooth properly increases as more care, skill, and judgment are applied to these early steps in restoration construction. When used as described above with any of the trays mentioned, the dual arch impression technique can be of great value in restorative dentistry. PMID- 16366045 TI - Avoiding supervised neglect. PMID- 16366046 TI - Eat this way. PMID- 16366047 TI - Split increment horizontal layering: a simplified placement technique for direct posterior resin restorations. AB - A new simplified technique for restoring moderate-to-large Class I occlusal cavities has been proposed. This technique utilizes split increment horizontal placement of composite resin, combined with shade or chroma stratification. It is believed that this technique will minimize the detrimental effects of polymerization shrinkage on the adhesive interface and cavity walls. PMID- 16366048 TI - Antimicrobial resistance of Staphylococcus aureus and oral streptococci strains from high-risk endocarditis patients. AB - This study sought to determine the antimicrobial susceptibility of Staphylcoccus aureus and viridans group streptococci strains collected from the forearm skin and saliva of 30 patients at high risk of endocarditis. Agar susceptibility tests of antibiotics routinely utilized in dentistry were used to verify antimicrobial resistance of bacterial strains. Of the Staphylcoccus aureus strains, 50% were resistant to ampicillin, 53.3% to amoxicillin, 60.0% to penicillin G, 13.3% to amoxicillin/clavulanate, 20.0% to azithromycin, 27.6% to clarithromycin, 23.3% to erythromycin, 3.3% to cefazolin, and 6.7% to clindamycin. Regarding streptococci, 16.7% of the strains were resistant to ampicillin, 16.7% to amoxicillin, 23.3% to azithromycin, 23.3% to clarithromycin, 30.0% to erythromycin, 13.3% to cefazolin, 26.7% to clindamycin, 16.7% to penicillin G, and 3.3% to amoxicillin/clavulanate. Pathogens associated with bacterial endocarditis exhibited elevated resistance rates against the antibiotics used for prophylaxis in dentistry. PMID- 16366049 TI - Using a diode laser to uncover dental implants in second-stage surgery. AB - A soft tissue diode laser offers an alternative technique for uncovering dental implants. This article presents two cases in which four dental implants were uncovered using a soft tissue laser. This technique provides an efficient and patient-friendly method to perform second-stage implant surgery, safely allowing a faster rehabilitative phase. PMID- 16366050 TI - Intraseptal anesthesia: a review of a relevant injection technique. AB - Although overshadowed by intraosseous anesthesia and the periodontal ligament injection, intraseptal anesthesia remains a useful local anesthesia technique for general dentists. Intraseptal anesthesia can be employed with safety and efficacy as an alternative to conventional local infiltration or regional nerve block injections. It also can serve as an adjunctive technique when conventional techniques fail to achieve adequate local anesthesia. This article reviews the intraseptal anesthesia technique, including its indications and limitations. PMID- 16366051 TI - Parotid duct sialolithiasis in a patient with Down syndrome--case report. AB - Sialolithiasis is the phenomenon of blockage of the salivary glands. It frequently is associated with swelling, pain, and infection of the affected gland. Clinically, sialolithiasis manifests as an increase in the size of the affected gland and increased salivary secretion that results in pain during eating. It occurs mainly in the submandibular gland and less frequently in the parotid gland. This article presents a case involving a 23-year-old woman with Down syndrome who demonstrated sialolithiasis in the parotid duct gland. The sialolith was radiographed and removed surgically. PMID- 16366052 TI - Success or failure of dental implants? A literature review with treatment considerations. AB - This study reviews the literature concerning the success or failure of dental implants and provides the general dentist with information to decide whether to recommend dental implant therapy to a patient. The authors conducted an extensive literature search for articles relating to dental implant failure. Metaanalyses and multi-center studies were predominant in the selection. Predictors of dental implant success or failure were gleaned from various articles and presented in the form of text and tables. The main predictors for implant success are the quantity and quality of bone, the patient's age, the dentist's experience, location of implant placement, length of the implant, axial loading, and oral hygiene maintenance. Primary predictors of implant failure are poor bone quality, chronic periodontitis, systemic diseases, smoking, unresolved caries or infection, advanced age, implant location, short implants, acentric loading, an inadequate number of implants, parafunctional habits and absence/loss of implant integration with hard and soft tissues. Inappropriate prosthesis design also may contribute to implant failure. PMID- 16366053 TI - Dental implants in the HIV-positive patient--case report and review of the literature. AB - The association between HIV infection and the success of osseointegrated dental implants is unknown. Recent reports in the dental literature show no difference between HIV-positive and HIV-negative patients in terms of infection rate after surgical procedures. A case report documenting the successful placement and osseointegration of eight endosseous dental implants in an HIV-positive patient is presented. This article also reviews the complications of implant procedures and the outcomes of surgical procedures in HIV-positive patients. PMID- 16366054 TI - Microleakage within endodontically treated teeth using a simplified root canal preparation technique: an in vitro study. AB - This study evaluated the effects of two canal preparation methods on leakage in endodontically treated teeth, comparing the relative leakage rates for conventionally prepared teeth and those instrumented and obturated with the Ultradent Endo-Eze system. Two groups (n=11) of freshly extracted human premolars (and some canines) with closed apices and single roots were used. The roots in the first group were instrumented with conventional stainless steel hand files and obturated with gutta-percha and Sealapex. The experimental group were instrumented and obturated using the Ultradent Endo-Eze system. Radiographic examination showed no difference in the level or quality of root canal fill for teeth prepared conventionally and those prepared using the Endo-Eze system. Leakage followed a linear trend for both groups of teeth but the rate of leakage increase was approximately twice as high (that is, 80% greater) for the conventional group than for the Endo-Eze teeth. This difference was not statistically significant due to the wide variability in the conventional group data. It was noted that canal preparation time for the Endo-Eze teeth was approximately 50% shorter than that required for conventional root canal therapy. PMID- 16366055 TI - Exophytic and ulcerated nodule of tongue. Pyogenic granuloma. PMID- 16366056 TI - Evidence and context. PMID- 16366057 TI - International challenges in health--can we succeed? PMID- 16366058 TI - Outbreak of campylobacteriosis following pre-cooked sausage consumption. AB - OBJECTIVES: A small outbreak of campylobacteriosis involving three cases was investigated in terms of Campylobacter types present in the suspect food (pre cooked sausages) and clinical samples from the cases. METHOD: Foods and faecal samples from people involved in the incident, which occurred in Christchurch, New Zealand, were tested for the presence of Campylobacter and identification of the species made. Isolates were typed by Penner serotyping and macrorestriction analysis using pulsed-field gel electrophoresis (PFGE). Investigations were conducted as to whether the contamination was on the surface or the interior of the sausages. RESULTS: All isolates from food and faecal samples were identified as C. jejuni and were indistinguishable from one another by the typing methods employed. Only the surfaces of the sausages were contaminated. Three other isolates of an indistinguishable subtype were isolated from campylobacteriosis cases in Christchurch occurring over approximately the same period. CONCLUSIONS AND IMPLICATIONS: Given the rarity of the subtype isolated from the three family members and the three other cases, it is possible that the outbreak was larger than the initial investigation revealed. It is likely that the sausages were contaminated after they had been cooked by the retailer and were not reheated prior to consumption. This report illustrates the role of cross-contamination in an outbreak with an unusual food vehicle for campylobacteriosis. Physical separation of cooked and raw product is necessary to prevent recurrences of outbreaks similar to the one described here. PMID- 16366059 TI - A cluster of thyrotoxicosis associated with consumption of a soy milk product. AB - OBJECTIVE: To determine whether a New Zealand cluster of thyrotoxicosis with low uptake on scintiscan was associated with soy milk consumption. METHOD: A case control study was conducted, with controls matched by age, sex, and region. RESULTS: All cases (5/5) and one control (1/30) consumed soy milk before the index date. The cases all drank a brand of soy milk that had unexpectedly high levels of iodine. In a simple unmatched unadjusted analysis, the lower bound of the 95% confidence interval for the odds ratio was 19. CONCLUSION: This cluster was associated with consumption of a soy milk product. The thyrotoxicosis was probably due to iodine-induced thyrotoxicosis. IMPLICATIONS: This investigation raises issues about quality control in manufacturing processes and the monitoring of food products for their safety. It also raises issues about public and health professional awareness of the potential health effects from food additives in some processed foods. PMID- 16366060 TI - The public health implications of a sporadic case of culture-proven Legionnaires' disease. AB - Legionella pneumophila is an important cause of community-acquired pneumonia. Domestic sources of infection have been increasingly recognised among community acquired cases. This report summarises the public health investigations and management of a single community-acquired case of Legionnaires' disease in Queensland, commenced in the context of a suspected outbreak. Legionellae from the case's domestic water supply were indistinguishable from the clinical isolate. The implications for future investigation of sporadic cases are discussed. PMID- 16366061 TI - Soil as a source of Legionella pneumophila serogroup 1 (Lp1). AB - OBJECTIVE: To investigate the potential source of a case of Legionnaires' disease caused by an unusual serotype of Legionella pneumophila serogroup 1 (Lp1) in regional Victoria in May 2001. METHOD: Epidemiological and environmental investigation of the source of infection of a case of Legionnaires' disease in regional Victoria in May 2001. RESULTS: Extensive environmental investigations did not reveal any cooling water tower systems close to the residence or the shopping centre that the case visited prior to illness. The sputum culture and a soil sample from the field at the plant nursery where the case worked prior to illness were both positive for Legionella pneumophilia serogroup 1, MDU pulsovar 97:103. CONCLUSION: Legionella pneumophila has been found in soil and was further found to be associated with a case of Legionella pneumophila. IMPLICATIONS: Public health authorities should consider exposures to soil and potting mixes when investigating cases of Legionella pneumophila where the case has no apparent association with cooling towers. Safe gardening practices should be promoted among the community. PMID- 16366062 TI - Assessment of a register-based rheumatic heart disease secondary prevention program in an Australian Aboriginal community. AB - OBJECTIVE: To assess specific performance indicators relating to a register-based acute rheumatic fever and rheumatic heart disease (ARF/RHD) prevention program in a remote Australian Aboriginal community in order to identify the most appropriate avenues for improvements in delivery of services. METHODS: Information kept on the central ARF/RHD register was compared with an amalgamated dataset from three other sources. The community clinic charts of identified patients were reviewed for information regarding accuracy of diagnosis and the number of doses of benzathine penicillin received in the last year. Specific follow-up arrangements were assessed and compared with practice guidelines. RESULTS: The central ARF/RHD register contained the names of 58 of the 72 (81%) people identified in the community as eligible for inclusion. Only 42% (22/52) of people receiving antibiotic prophylaxis had received 80% or more of the recommended doses in the previous year; service delivery was significantly better for females than males (p = 0.004). Individuals in priority category 1 (most severe disease) were found to be receiving follow-up and investigation according to guidelines. About half the people in categories 2 (moderate disease) and 3 (mild disease) had been inadequately investigated and/or missed out on follow-up appointments. CONCLUSIONS: The ARF/RHD prevention program in this large remote Aboriginal community is struggling to deliver services to a substantial proportion of people who require them. Specific interventions, especially those related to men's health, may be required to correct the problems. PMID- 16366063 TI - Use of the Jones Criteria in the diagnosis of acute rheumatic fever in an Australian rural setting. AB - OBJECTIVE: To answer the question, are the Jones Criteria being used appropriately in the diagnosis of acute rheumatic fever (ARF) by non-specialist medical staff in a remote Australian setting? METHODS: The medical records of all patients discharged from Katherine Hospital (Northern Territory) with a diagnosis of ARF between January 2000 and April 2004 were retrospectively reviewed for adherence to the Jones Criteria. Data were also collected on specialist follow-up and need for transfer to a tertiary hospital. RESULTS: Twenty-five patients had a diagnosis of ARF and all were Aboriginal or Torres Strait Islander. Thirty-two per cent did not fulfil the Jones Criteria and of these 63% were recurrent cases. Eighty-eight per cent received specialist follow-up and of those who did not fulfil the Jones Criteria, all were diagnosed as ARF by the specialist. Only 20% required transfer to a tertiary hospital for higher-level care. CONCLUSION: The Jones Criteria are being used appropriately to diagnose initial episodes of ARF but less successfully in recurrent episodes. Specialist follow-up is essential but acute episodes can be managed in remote settings, reducing the need to transfer patients to tertiary care with resultant patient dislocation and social isolation. IMPLICATIONS: The diagnosis of ARF results in long-term penicillin prophylaxis. This is a major public health undertaking that requires correct diagnosis. This study demonstrates that the Jones Criteria are being used appropriately to diagnose ARF in a remote setting. The ability to diagnose and treat Indigenous patients within their local region reduces social isolation and creates a more positive health care experience. PMID- 16366064 TI - Estimating influenza vaccine effectiveness in an outbreak when anti-viral medications were used as a control measure. AB - OBJECTIVES: To estimate influenza vaccine effectiveness (VE) from an outbreak of influenza A in an aged care facility in which neuraminidase inhibitors were used as part of outbreak control measures. METHODS: The outbreak occurred between 25 December 2001 and 21 January 2002. Neuraminidase inhibitors were used to control the outbreak. Residents and staff with respiratory symptoms were tested for influenza using RT-PCR and/or serology. Vaccine effectiveness (VE) was estimated for the prevention of laboratory-confirmed influenza. RESULTS: Nineteen of 42 (45%) residents and four of 29 (14%) staff were laboratory-confirmed as cases. The outbreak was caused by an influenza A (H3N2) strain, antigenically matched to that season's vaccine. The VE for preventing laboratory-confirmed influenza infection was 61% (95% CI 41-73) among residents and 100% (95% CI 63-100) among staff. CONCLUSIONS: The VE estimates calculated from this outbreak were consistent with other published results. Outbreaks of influenza in institutions provide a good opportunity to review influenza VE, but use of antiviral medications as control measures may affect interpretation of findings. PMID- 16366065 TI - Modelling the impact of vaccination on the epidemiology of varicella zoster virus in Australia. AB - OBJECTIVE: To model the impact of universal varicella vaccination in Australia. METHODS: The results of an Australia-wide serosurvey for varicella zoster virus (VZV) immunity were used to parameterise realistic, age-structured deterministic models (RAS) developed by Brisson and colleagues. We examined the impact of a vaccination program for one-year-olds alone, and with a catch-up campaign for 11 year-olds, on the incidence of varicella and zoster, using Australia's population structure. Morbidity was then determined by calculating the number of hospital in patient days. RESULTS: Infant vaccination is predicted to reduce the incidence of varicella. However, zoster incidence is expected to increase initially, assuming exposure to varicella boosts immunity to zoster. Accumulated morbidity from both varicella and zoster is predicted to remain above that expected without vaccination for the first 70 years of an infant program (assuming 90% coverage with boosting for 20 years). However, after 70 years the net health savings from vaccination are predicted to increase substantially. CONCLUSIONS AND IMPLICATIONS: Infant vaccination is expected to be a successful long-term commitment to reducing morbidity associated with VZV infection in Australia. PMID- 16366066 TI - An evaluation of public health interventions during a pertussis outbreak in the ACT, 2003. AB - OBJECTIVE: To assess whether issuing information alerts increased the proportion of pertussis cases notified within the 21-day infectious period during a pertussis outbreak in the Australian Capital Territory (ACT) in 2003. METHOD: The study included all persons resident in the ACT for whom a notification of pertussis was received with onset of disease between 1 March and 31 December 2003. Cases notified within 21 days of onset were called 'early notifications'. The effectiveness of information alerts was assessed by comparing the proportion of early notifications before and after each alert and over the course of the outbreak. MAIN OUTCOME MEASURE: The proportion of early notifications. RESULTS: The proportion of early notifications ranged from 1/36 (2.7%) prior to the first intervention to 29/52 (55.7%) after the final intervention (p < 0.001). CONCLUSIONS: This study supports the hypothesis that information alerts by ACT Health increase the proportion of cases notified within the infectious period of 21 days. IMPLICATIONS FOR PUBLIC HEALTH PRACTICE: Early diagnosis of pertussis cases (within the infectious period) has the potential to limit transmission and protect susceptible infants. PMID- 16366067 TI - Correct procedures and cutting corners: a qualitative study of women's occupational health and safety in a beauty therapy industry. AB - OBJECTIVE: To examine the context of occupational health and safety related to blood-borne communicable diseases practice. METHODS: A case study approach using qualitative semi-structured interviews with five key informants who represented different sectors of the beauty therapy industry in South Australia. RESULTS: Four main themes were identified: (i) exposure to blood and blood-borne communicable diseases; (ii) prevention in practice; (iii) OH&S problems; and (iv) industry needs. CONCLUSION: Key OH&S issues in the beauty therapy industry include: power relationships between employers and employees, equipment costs, the need for more continuing education, and monitoring of practitioners. IMPLICATIONS: Economic constraints, continuing education, and government regulation of the beauty therapy industry are highlighted as significant areas for further consideration in addressing the OH&S needs of practitioners and their clients. PMID- 16366068 TI - Health professionals' knowledge, practice and opinions about fetal alcohol syndrome and alcohol consumption in pregnancy. AB - OBJECTIVE: To measure the knowledge, attitudes and practices of health professionals regarding fetal alcohol syndrome (FAS) and alcohol use during pregnancy. METHOD: A postal survey of a representative random sample of health professionals was conducted in Western Australia (WA) in 2002/03. 1,143 (79%) of 1,443 eligible health professionals completed the survey (87 Aboriginal Health Workers, 286 allied health professionals, 537 community nurses, 170 general practitioners and 63 obstetricians). RESULTS: Of 1,143 health professionals, 12% identified all four essential diagnostic features of FAS. Most (95%) had never diagnosed FAS. Although 82% believed that making a diagnosis of FAS might improve treatment plans and 85% agreed FAS was preventable, 53% said the diagnosis might be stigmatising. Only 2% felt very prepared to deal with FAS and most wanted information for themselves and their clients. Of the 659 health professionals caring for pregnant women, only 45% routinely ask about alcohol use in pregnancy, only 25% routinely provide information on the consequences of alcohol use in pregnancy and only 13% provide advice consistent with NHMRC guidelines on alcohol consumption in pregnancy. CONCLUSION: Health professionals have identified the need for educational materials for themselves and their clients. IMPLICATIONS: FAS is likely to be under-ascertained in Australia due to a lack of knowledge of FAS by health professionals. Until this lack of knowledge is addressed, opportunities for diagnosis and prevention of FAS will be limited. PMID- 16366069 TI - How to calculate standard errors for population estimates based on Australian National Health Survey data. AB - The National Health Surveys (NHS), conducted regularly by the Australian Bureau of Statistics, are designed to obtain benchmark information on a range of health related issues and to enable the monitoring of health trends over time. Confidentialised Unit Record Files (CURFs), containing detailed individual-level NHS data, are available from the ABS. When using the CURF data to make an estimate relating to the Australian population, the precision of the estimate should also be calculated, but due to the complexity of the survey design and the limited survey design information in the CURF, this is not straightforward. This paper explains three different methods for calculating standard errors for population estimates obtained using CURF data: (i) using the estimates of relative standard errors published by the ABS; (ii) using the replicate weights provided by the ABS in the CURF; and (iii) assuming the survey design is a simple random sample. The most appropriate method will depend on which survey is being analysed and the type of population estimate; this is discussed in detail. Worked examples for each method using data from the 2001 NHS CURF are provided, together with Stata 9 code illustrating the implementation of each method. PMID- 16366070 TI - Mortality among prisoners: how accurate is the Australian National Death Index? AB - OBJECTIVE: The effectiveness of the National Death Index (NDI) in identifying deaths among different populations has been studied before. Prisoners represent a segment of the population that might be expected to be difficult to link. The objective of this research was to evaluate the accuracy of the Australian NDI to ascertain the mortality status of prisoners in New South Wales, Australia. METHODS: Data on 311 inmates who had died in custody between January 1988 and March 2003, and on 7,558 inmates known to be alive at the end of May 2003, were matched to the NDI database. Matching was undertaken using a probabilistic linkage method followed by a clerical review. The underlying causes of death reported by the NDI were compared against those reported by the NSW Coroner and held at the Legal Branch of the Department of Corrective Services. RESULTS: Linkage correctly identified 275 of the 311 known deaths (sensitivity: 88.4%). Of the 7,558 prisoners known to be alive in custody at the end of May 2003, linkage to the NDI incorrectly identified 23 as dead (specificity: 99.7%). Cause of death was accurately reported by the NDI for 95.5% of deaths identified by the linkage. CONCLUSION: The results of this study show that the NDI is an accurate and feasible means for evaluating mortality among prisoners. The nature of the prisoner population makes the linkage a difficult task for the NDI. The success of the data linkage depends on the extent and accuracy of information contained in the index file and type and quality of information available. PMID- 16366071 TI - Do university ethics committees adequately protect public health researchers? AB - OBJECTIVE: This paper aims to examine whether university human research ethics committees (HRECs) proactively seek to protect members of the research team as well as study subjects in their written documentation. METHODS: A content analysis of 37 Australian university HREC application forms and attachments was undertaken. Each form was allocated to one of four predetermined categories. RESULTS: Of the 37 forms, only three included an explicit request for the applicant to reflect on all possible aspects of safety of the researchers (physical, psychological and emotional). CONCLUSION: Few HRECs have taken issues of possible harm to researchers into account in their documentation. It is recommended that HRECs explicitly recognise potential risks to researchers, especially those engaged in exploration of sensitive topics, in their processes of approving human research. It is also recommended that researchers consider the possible implications of undertaking this type of research and ensure strategies are in place to minimise these risks. PMID- 16366072 TI - Sample size is beside the point in policy development research. PMID- 16366073 TI - Feeling bad about immunising children. PMID- 16366074 TI - The case: Transient patellar dislocation. PMID- 16366075 TI - A white man's take on diversity. PMID- 16366076 TI - Smart orthopedic implants. PMID- 16366077 TI - A new technique of crossed pins in supracondylar elbow fractures in children. PMID- 16366078 TI - The effect of volume and pressure on femoral cement grade. PMID- 16366079 TI - Staying in rhythm: identifying risk factors for Torsade de pointes. PMID- 16366080 TI - Leg salvage after gunshot wound to the distal femur: patience is a virtue. PMID- 16366081 TI - Unicondylar knee arthroplasty: past and present. PMID- 16366082 TI - Acetabular reconstruction in revision hip surgery using femoral head block allograft. AB - This prospective study analyzed the clinical and radiological results of 140 consecutive cases of acetabular revision using large frozen femoral head allografts and cemented all-polyethylene acetabular components. Mean follow-up was 10 years (range: 5-16 years). Thirty patients died, seven were lost to follow up, and 26 had failed and undergone further surgery. Nineteen failures were due to aseptic failure and collapse of the graft. Kaplan-Meier survival analysis calculated a mean survival at 10 years of 88.5% for revision for any reason. We compared all reported techniques of acetabular reconstruction for similar defects and recommend a surgical strategy based on the available evidence, but weighted towards a preference to reconstitute bone stock rather than removing further bone in the revision situation. PMID- 16366083 TI - Extramedullary versus intramedullary tibial alignment guides for total knee arthroplasty. AB - This retrospective study evaluated whether intramedullary or extramedullary tibial alignment guides are more accurate in assuring correct tibial component positioning in total knee arthroplasties (TKAs). Fifty-five TKAs were performed with the only difference being whether an intramedullary (31) or extramedullary (24) cutting guide was used. Pre- and postoperative tibiofemoral angle, tibial component alignment angle, and femoral component alignment angles were measured and the results were statistically analyzed. We also performed a subgroup analysis on the preoperative varus knees to determine if the presence of a varus deformity would have better results with either the intramedullary or extramedullary cutting guide. The 2-sample t test was used to compare the results of the two groups. The results showed no statistical difference in alignment of the tibial component when using the intramedullary or extramedullary guides. PMID- 16366084 TI - The reverse sural neurocutaneous flap for hindfoot and ankle coverage: experience and review of the literature. AB - Soft-tissue defects about the ankle are common in orthopedic surgery. The wounds usually involve exposed bone or hardware, making skin grafting a poor option. Free tissue transfer remains a mainstay of treatment. Recently, the reverse sural neurocutaneous flap has gained popularity. At our institution, 11 fasciocutaneous flaps have been performed for defects averaging 53cm2. All flaps achieved stable coverage. One patient required repeat skin grafting. Given its reliability, easy dissection, and outpatient capability, this flap should be considered for coverage in this region. PMID- 16366085 TI - Total hip arthroplasty with satin finish, tapered stems. AB - Subsidence of femoral hip prostheses with a rough surface has been associated with osteolysis and loosening. However, recent evidence has suggested that smooth finish tapered stems may not incur these problems. An experimental monobloc satin finish, tapered femoral component was designed to subside within the cement. There were 49 femoral components implanted, and patients averaged a 1.8-mm subsidence at the prosthesis-cement interface with a follow-up of 6.76 years. There were no failures attributable to aseptic loosening at the cement-bone interface. This study supports the use of a satin finish, tapered femoral component in cemented total hip arthroplasty. PMID- 16366086 TI - Vancomycin-resistant Enterococcus faecium osteoarthritis following Staphylococcus aureus hip infection. PMID- 16366087 TI - Reconstruction of a true ulnar artery aneurysm in a 4-year-old patient with radial artery agenesis. PMID- 16366088 TI - Simultaneous fractures of all seven cervical vertebrae. PMID- 16366089 TI - Genetic polymorphism in Plasmodium falciparum vaccine candidate antigens. AB - Malaria is still a major public health problem in many tropical and subtropical countries. Malaria vaccine is highly desirable as an adjunct to existing malaria control measures. The polymorphisms in malaria vaccine candidates antigens might be a hurdle in developing an effective vaccine. The present article reviews the genetic polymorphism in several antigens expressed on the parasite surface, which are targets for immunological responses of the host and are good candidates for vaccine development against P. falciparum. Variable regions of most genes are generally dimorphic probably as a result of intragenic recombinations. Each allele in turn shows polymorphism resulting from point mutations, or other mechanisms. Several antigens like merozoite surface protein-1 and 2 (MSP-1 and MSP-2) and S antigen show high polymorphism while in others like circumsporozoite protein (CSP), apical membrane antigen-1 (AMA-1) and erythrocyte binding antigen 175 (EBA-175) functional constraints limit the degree of polymorphism. Polymorphism reported in these genes is discussed. PMID- 16366090 TI - Clinicopathological prognostic factors of adult granulosa cell tumor of the ovary -a study of 37 cases. AB - Adult granulosa cell tumor (GCT) of the ovary is the most common malignancy amongst the sex cord stromal tumors. Clinical stage, age, tumor size, residual disease and several histologic factors have been reported to be of prognostic significance. There were 839 ovarian malignancies at our institute during the period from 1998 to 2002. Out of 54 granulosa cell tumors, 40 cases were evaluable which includes 37 cases of adult GCT and 3 cases of juvenile GCT. They were studied retrospectively. Majority of patients presented with abdominal symptoms and FIGO stage I. All patients were treated surgically and 62.1% of adult GCTs were given post-operative chemotherapy. Clinical stage, presence of residual disease and tumor volume were the most important prognostic factors. Age of patient, menstrual status, post-operative chemotherapy, mitosis or histological patterns were of little significance in our study. PMID- 16366091 TI - Significance of apoptosis in prostatic intraepithelial neoplasia and carcinoma. AB - This study was carried out to assess the significance of apoptosis in prostatic intraepithelial neoplasia (PIN) and prostate cancer. A total of 120 prostatic specimens were studied in the department of Pathology [corrected] JNMC, Aligarh. The rate of apoptosis in PIN and prostate cancer was examined by quantifying the number of apoptotic bodies per hundred cells (apoptotic index) on haematoxylin and eosin stained histological sections [corrected] A significant correlation was noted between increasing apoptotic indices and increasing Gleason grades within a cancer. PMID- 16366092 TI - Invasive fungal infections in renal allograft recipients. AB - Invasive fungal infections contribute significantly to morbidity and mortality in renal allograft recipients. We identified 29 cases of invasive mycoses on histological and/or cytological examination, out of the total 1231 renal transplants performed at our centre over a period of last 15 yrs (1989-2003). A detailed clinical analysis was performed. The time interval between transplant and the occurrence of invasive fungal infection ranged from 15 days to 10.5 yrs. Candida and Aspergillus were the most frequent offenders (66%); Candida alone accounting for 45% of the cases. The most common risk factors were post transplant Cytomegalovirus infection, diabetes mellitus and episodes of acute rejection. Fine needle aspiration cytology, bronchoalveolar lavage and esophageal brush smears aided in prompt diagnosis. Disseminated infection was associated with a high mortality (80%). Management of renal transplant recipients requires identification of risk factors and early clinical suspicion of infection. The role of prophylaxis needs further evaluation. PMID- 16366093 TI - Glomerular basement membrane thickness in normal adults and its application to the diagnosis of thin basement membrane disease: an Indian study. AB - Microscopic hematuria of non-urologic origin warrants ultrastructural study of renal biopsy. Thinning and variations in the texture of glomerular basement membrane (GBM) are difficult to be recognized under light microscope; transmission electron microscope (TEM) therefore plays a vital role in identifying such changes. Ultrastructural morphometry is a valuable diagnostic aid when GBM is suspected of being abnormally thin. In an effort to determine the normal GBM thickness (GBMT) in Indian adults and to determine the cutoff value of GBMT for a diagnosis of thin basement membrane disease (TBMD), we determined GBM thickness in 25 normal adults. Postmortem biopsies of 25 normal adults (16 males and 9 females) aging between 18-58 years were included in the study. GBM thickness was determined through ultrastructural morphometry on accurately enlarged electron micrographs as harmonic mean of 50 orthogonal intercepts across the GBM in each case. Study revealed a mean GBM thickness of 321 nm with a standard deviation (SD) of 28 nm. Mean-2SD (321-56), that is 265 nm, was fixed as cutoff value of GBMT for the diagnosis of TBMD. A systematic split study of control subjects revealed thicker GBM (329+/-38 nm) in higher age group (35-60 years) as compared to GBMT (316+/-21 nm) in lower age group (18-30 years). Males in higher age group also revealed thicker GBM (males: 343+/-39 nm versus females: 300+/-12 nm). Ten patients with non-urologic hematuria and having GBMT<265 nm were diagnosed as cases of TBMD. Patients with TBMD revealed significantly attenuated GBM as compared to age and sex matched controls (214 +/- 40 nm versus 311 +/- 17 nm; p<0.0005). No overlap was observed in the distribution of GBMT in patients of TBMD and age and sex matched controls. Ultrastructural morphometry is the ultimate and appropriate method for diagnosing TBMD. PMID- 16366094 TI - Renal angiomyolipomas--a study of 18 cases. AB - Angiomyolipoma is a distinctive neoplasm composed of an intimate admixture of three components, viz. mature adipocytes, smooth muscle cells and blood vessels. This study was undertaken to better define the various morphological patterns of angiomyolipoma and their immunohistochemical profile. The paraffin blocks and slides of 18 cases of renal angiomyolipoma, accessioned over a period of 8 years from Tata Memorial Hospital, were reviewed. There were 2 men and 16 women in the age range of 17 to 68 years. Pre-operative fine needle aspiration cytology (FNAC) was performed in 6 cases, of which 5 were erroneously diagnosed as renal cell carcinomal sarcoma. Histologically, 14 cases revealed conventional histology. The remaining 4 cases were particularly misleading posing diagnostic problems due to variant patterns (leiomyomatous variant-3 cases and epithelioid variant- 1 case). Apart from the usual histology, necrosis, giant cells, and varying degrees of nuclear pleomorphism were present in three of the cases. Immunohistochemistry performed in 16 cases revealed immunoreactivity for HMB45 in all the 16 tumors (100%), estrogen receptor (ER) and progesterone receptor ((PR) in 2 cases (12.5%) and 6 cases (37.5%) respectively. PMID- 16366095 TI - Comparative study of microscopic detection methods and haematological changes in malaria. AB - Quantitative buffy coat (QBC) technique, thick smears, thin smears and conventional buffy coat smears were compared for malarial parasite detection. Of 200 blood samples studied, 70 (35%) samples were positive by QBC technique, 62 (31%) samples by thick smears, 50 (25%) samples by thin smears and only 34 (17%) samples were positive by conventional buffy coat technique. 8 (4%) samples detected by QBC technique alone, had low parasitic index. Haematological analysis was performed on samples positive for malarial parasite. Anaemia was present in 66 (94.28%) samples of which 37 (56.06%) were Plasmodium falciparum, 21 (31.81%) were Plasmodium vivax and 8 (12.12%) had mixed infection (Plasmodium falciparum and Plasmodium vivax). 35 (50%) cases showed normocytic normochromic anaemia. Majority of the samples showed normal total and differential leukocyte count. Thrombocytopenia was found in 49 (70%) samples of which 33 (67.34%) were Plasmodium falciparum. PMID- 16366096 TI - Meningiomas: report of a hospital-based registry. AB - This is a hospital-based epidemiologic study of meningiomas. Of 1321 central nervous system tumours, meningiomas constituted 21% of the cases, being the second largest category of a single histologic type after astrocytomas. Of the 267 meningiomas studied, 247 were intra-cranial (92.5%). The age of the patients varied between 6 to 84 years. Histological subclassification is presented and treatment schedules discussed. 261 (98%) meningiomas were histologically benign and 5 were malignant meningiomas (1.9%). A 5-year follow-up was available in most cases, with the help of which it was possible to understand the biological behaviour of various sub-types and the influence of other parameters such as location and treatment schedules. Of note was the fact, that out of 261 patients with benign meningiomas, 11 succumbed in the immediate post-operative period and in 8 of these cases, the tumour was located at the base of the skull. PMID- 16366097 TI - Disseminated Strongyloides stercoralis in AIDS: a report from India. AB - We report a fatal case of disseminated strongyloidiasis masquerading clinically as stage IV caecal malignancy diagnosed at post mortem by needle necropsy. The parasite was seen in the smears from CSF, pleural fluid, ascitic fluid, splenic aspirate, lung aspirate and aspirates from caecal area. Enteric organisms like Group D streptococci and candida sp were also associated. We believe that this is the first report of widespread dissemination of S. stercoralis in AIDS from India. PMID- 16366098 TI - Congenital fibrosarcoma of the upper extremity--a case report and review of literature. AB - Fibrosarcoma is rare in pediatric age group. Its diagnosis in infants may be difficult clinically and histologically. A twenty-day infant presented with a rapidly growing mass in the right forearm. Histologically a gray white fleshy mass was seen occupying the entire forearm and distal arm. The tumor showed sheets of closely packed spindle cells with minimal pleomorphism. The cells were positive for vimentin, negative jor smooth muscle actin, desmin and S-100. The diagnosis of congenital fibrosarcoma was made. Most cases of infantile fibrosarcomas present in the first year of life and although they have an alarming presentation, in children they have a much better prognosis as compared to adults. Diagnosis can be made on histology with immunohistochemistry. PMID- 16366099 TI - Lipomatous metaplasia occurring within meningiomas--two case reports. AB - Lipomatous meningiomas have recently been described as rare well defined variants of meningiomas characterized by meningothelial cells which undergo a metaplasia to adipocytes. Presence of intratumoral fat can sometimes cause confusion in preoperative radiological diagnosis. We report two such histologically unusual cases of meningiomas with significant adipose tissue metaplasia. PMID- 16366100 TI - Tay Sachs disease: an autopsy case report. AB - This report describes a case report of a postmortem performed on a 5-year old patient of Tay-Sachs disease, presenting with failure to thrive, muscular flaccidity, and cherry-red spots on macula on fundoscopy. There was no history of similarly affected sibling or any other family member. The diagnosis was confirmed by enzyme studies. At postmortem, there was no organomegaly. The brain, on microscopy, showed vacuolated swollen neurons. PMID- 16366101 TI - Clear cell carcinoid tumour of lung--a case report. AB - An unusual case of pulmonary clear cell carcinoid tumour in a fifteen year old boy is being described. The differential diagnoses considered in this case are described. This is one of the uncommon varieties of carcinoid of lung reported in the world literature. PMID- 16366102 TI - Subcutaneous metastases of sarcomatoid mesothelioma with its differential diagnosis on fine needle aspiration--a case report. AB - Metastasis of mesothelioma of the pleura, to the skin and subcutis is an extremely rare occurrence. A 25 year old woman, who had undergone chemotherapy, partial excision of tumor followed by radiotherapy of sarcomatoid mesothelioma of the pleura, presented three months later with painless widespread subcutaneous nodules. FNAC of these nodules reveled pleomorphic malignant spindle shaped cell with epithelioid morphology. The subcutis is a particularly rare site of metastatic sarcomatoid mesothelioma. It is essential to differentiate neoplasm metastatic to the skin and subcutis from primary and benign lesions of the same region. FNAC is accurate and efficient, in conjugation with clinical history, and it also prevents surgical biopsy in the diagnosis of metastatic subcutaneous lesion. To our knowledge, this is the first case, reported till date, in which the sarcomatoid mesothelioma metastasized to the subcutaneous tissue and was diagnosed by fine needle aspiration cytology (FNAC). PMID- 16366103 TI - Fatty acid oxidation disorder as a cause of sudden infant death syndrome (SIDS)- a case report. AB - Although the cause of sudden infant death syndrome (SIDS) remains unknown, extensive studies over the last 10 years have begun to shed some light on this family tragedy. 5% of all cases of SIDS are caused by fatty acid oxidation disorders. We report a case of fatty acid oxidation disorder causing SIDS. PMID- 16366104 TI - Childhood hepatic angiosarcoma--a case report. AB - Hepatic angiosarcoma (HAS) is an extremely rare liver tumor in children. We report a case of childhood HAS in a six year old girl who presented with acute abdominal pain and fever with a mass in epigastrium. Left hepatic lobectomy was performed with a clinical diagnosis of hepatoblastoma. Histopathological examination revealed features typical of hepatic angiosarcoma. The case is presented for its rarity and to discuss the interrelation between infantile hemangioendothelioma (IHE) and HAS. PMID- 16366105 TI - Kikuchi-Fujimoto disease--a case report. AB - The etiology of cervical lymphadenopathy is multifactorial including infections and tumoral lesions. Histiocytic necrotising lymphadenitis or Kikuchi-Fujimoto's disease is a rare cause of cervical lymphadenopathy which does not have specific clinical signs. We report herewith a case of Kikuchi's disease (KD) diagnosed by histopathology. PMID- 16366106 TI - Cystic lymphangioma as mesenteric tumor--a case report. AB - Cystic lymphangioma is an uncommon mesenteric tumor usually reported in children. We report a case of 14 year old female who presented with dull aching abdominal pain. At laparotomy a cystic tumor was found in mesentery, which was attached to bowel loops. The histopathology examination showed features of cystic lymphangioma. Although pre-operative diagnosis is usually possible on CT or MRI, confirmatory diagnosis of this lesion requires laparotomy followed by histopathology. PMID- 16366107 TI - Sertoli-Leydig cell tumor of ovary with heterologous element: a case report. AB - Sertoli-Leydig cell tumors are uncommon tumors of ovary accounting for about 1% of sex cord stromal tumors. They constitute between 0.1% to 0.5% of all the primary ovarian neoplasms. The majority of reported cases are moderately or poorly differentiated combined Sertoli-Leydig cell tumors. A rare case of Sertoli Leydig cell tumor with heterologous element in a 14 year girl is described. The heterologous component comprised smooth muscle, an uncommon element, that was seen in 60 to 70% of the tumor area. The epithelial element of the tumor consisted of solid, tubular and foci of retiform pattern. Both these features imply a poor prognosis. PMID- 16366108 TI - Dedifferentiated papillary carcinoma of thyroid in an adolescent girl--a case report. AB - Dedifferentiated papillary carcinoma of thyroid shows combined histopathological features of classical papillary carcinoma and anaplastic carcinoma. This is a very rare tumour in adolescence. This prompted the authors to report this case. PMID- 16366109 TI - Angiosarcoma of scalp: a case report. AB - Cutaneous angiosarcomas are rare and constitute less than 1% of all malignant mesenchymal tumours. Most angiosarcomas of skin arise in the following clinical settings: 1. face and scalp of elderly 2. following lymphoedema and 3. post radiation. We present a case of an elderly man presenting with scalp lesion of 18 months duration. Histological examination of the biopsy revealed features of an angiosarcoma, which showed imunohistochemical positivity for factor VIII related antigen. PMID- 16366110 TI - Angioimmunoblastic T-cell lymphoma obscured by concomitant florid epithelioid cell granulomatous reaction--a case report. AB - Epithelioid cell granuloma occurs in association with many neoplasms including lymphoma. However they have rarely obscured the microscopic features of a lymphoma. We report on a case where a florid epithelioid cell granulomatous reaction caused difficulty in interpretation and delayed the final diagnosis of a case of peripheral T cell lymphoma of the angioimmunoblastic type. We draw attention to this unusual phenomenon because of its practical implications. PMID- 16366111 TI - Fine needle aspiration cytology of adenomatoid tumour--a case report with review of literature. AB - Adenomatoid tumours are neoplasms of male and female genital tract with the epididymis being the most common site. They also occur in uterus, fallopian tube, and ovary. These benign tumours are asymptomatic or cause mild symptoms and a palpable mass. Fine needle aspiration of these tumours is very useful to differentiate malignant from benign lesions and helps to avoid unnecessary aggressive surgical procedures. FNAC of these benign epididymal tumours is diagnostic, rapid, reliable, conclusive and cost effective. We are reporting a case of adenomatoid tumour of epididymis in a 41 year old male patient, diagnosed by FNAC and confirmed by histopathology. PMID- 16366112 TI - Placental site trophoblastic tumor. AB - Placental site trophoblastic tumor is very rare. It is a gestational trophoblastic neoplasm which follows normal or molar pregnancy. It is usually confined to uterus & 15-20% behave in a malignant fashion. Clinically patients present with abnormal excessive bleeding or amenorrhoea. Though several trophoblastic lesions & tumors enter in the differential diagnosis, microscopically the diagnosis is usually straightforward in hysterectomy specimens. Hysterectomy is the treatment with or without chemotherapy. Progress after surgery should be monitored by placental lactogenic hormone. PMID- 16366113 TI - Isolation of Enterococcus faecalis from a case of nonpuerperal breast abscess. AB - Nonpuerperal breast abscess (NPBA) has different etiology as compared to the mastitis occurring in post partum women. The condition presents either as acute suppurative infection or chronic type. Organisms usually implicated are Staphylococcus aureus, coagulase negative staphylococci, and anaerobes. Mostly the infection is polymicrobial in nature. Herein, we report the isolation of Enterococcus faecalis from a case of acute suppurative NPBA. PMID- 16366114 TI - FNAC in a case of NHL presenting initially as nodal infarction. AB - Lymph node infarction is rare and can occur in either nonneoplastic or neoplastic conditions. Fine needle aspiration cytology (FNAC) of infarction preceding lymphoma has not been described earlier. A 26-year-old male, was referred to the cytology laboratory for FNAC of bilateral axillary lymph nodes. FNA smears showed uniform looking ghost cells. There were no viable cells. A biopsy was advised which also showed extensive coagulative necrosis. Five weeks later, right cervical lymph nodes also appeared and FNA smears showed discrete monomorphic population of immature lymphoid cells. A cytologic diagnosis of infarction in a case of non-Hodgkin's lymphoma (NHL) was made and subsequently confirmed by histopathologic examination. Our case indicates that such cases should be followed up closely and repeated aspirations should be done to prevent a delayed diagnosis of lymphoma. PMID- 16366115 TI - Application of bleach method to improve sputum smear microscopy for the diagnosis of pulmonary tuberculosis. AB - In an active surveillance study, the bleach concentration method to improve sputum smear microscopy for the diagnosis of pulmonary tuberculosis was applied in chest symptomatics in Mahidpur block of Ujjain district in Madhya Pradesh, India. We purposely selected twenty villages with population of approximately 20,000 individuals. 664 sputum specimens from 297 chest symptomatics were collected. Ziehl-Neelsen staining was performed on direct sputum smears, smears made after bleach method and modified Petroff's method. Out of 297 chest symptomatics, 16 cases (5.38%) were positive by direct microscopy, 27 cases (9.09%) were positive by bleach method, 22 cases (7.40%) were positive by modified Petroff's method. The bleach method is safe, cheap, easy and sensitive. It can be applied for improved detection of Mycobacterium tuberculosis in hospitals and laboratories especially in settings where mycobacterial culture facilities are not available. The implementation of bleach method clearly improves case detection and can be a useful contribution in the National Tuberculosis Control Program. PMID- 16366116 TI - The seroepidemiological study on cytomegalovirus in women of child-bearing age with special reference to pregnancy and maternal-fetal transmission. AB - We screened 500 women of childbearing age belonging to different socioeconomic class for the presence of IgM antibodies against cytomegalovirus (CMV) infection by ELISA. Among these were 70 pregnant women, positive for CMVspecific IgM antibodies, whose newborns were also tested for the same. IgM positivity was found to be 5.4% (27/500) while 2.2% (11/500) gave equivocal results. There was an increasing trend in IgM positivity with age, decreasing socioeconomic status and increasing parity. Prevalence rate was more in women from rural as compared to those of urban area. Among the pregnant women, in the higher income group, 4 (14.28%) had CMV specific IgM antibodies as compared to 9 (21.43%) of low income. Congenital infection occurred more often (14.28 vs 7.14%) in infants in low income group. Signs and symptoms compatible with acute CMV infection were found in 7.69% (1/13) women and 27.78% (5/18) newborns. Thus the need for screening and protection against this infection is further emphasized. PMID- 16366117 TI - Standardization of parvovirus B19 DNA extraction from serum and quantitative PCR. AB - Human parvovirus B19 (B19) is a newly emerging virus causing a wide spectrum of clinical conditions. The corner stone of diagnosis of acute B19 infection is demonstration of anti-B19 IgM antibodies and its DNA by PCR. Sera of patients infected with B19 acutely or persistently shows intense viraemia (up to 10(12) virus particles/ ml) but extraction of B19 DNA from serum is a problematic task. There is no single standardized, cost-effective in-house method, which can successfully extract DNA of B19 from serum samples and which has been subsequently tested repeatedly by many investigators over time. We describe here an efficient in-house method of extraction of B19 DNA from serum and quantitate extracted DNA by PCR. Firstly, a quantitative PCR was done using 3 microl of a cloned B19 DNA (33.3 pg/ml) mixed in 47 microl of sterile distilled water which were further diluted from 10(-1) up to 10(-7) to find the lower limit of DNA detection. To mimic human serum samples with known quantity of B19, 3 ml of cloned B19 DNA (33.3 pg/ml) were mixed with 47ml of fetal calf serum (FCS; free of B19) and were similarly log diluted from 10(-1) to 10(-7) in 50 ml volume. In house method of DNA extraction from serum (FCS+B19 DNA) were then performed followed by quantitative PCR. In both the cases, we were able to detect B19 DNA up to 10(-4) dilution which contained 0.6 fg of B19 DNA corresponding to 12 B19 genome equivalents/PCR reaction or 2.4 x 10(2) genome quivalents/ml of serum. Further, the entire procedure was repeated two more times and identical results were obtained confirming its reproducibility. Using this in-house method we extracted and amplified B19 DNA successfully from sera of clinically suspected cases of B19 infection (data not shown). Compared to other studies, the sensitivity of our in-house method was found to be superior hence recommended for developing countries as commercial kits are too costly. PMID- 16366118 TI - Is Campylobacter involved in antibiotic associated diarrhoea? AB - Campylobacter jejuni is an important cause of acute bacterial diarrhoea. In developing countries like India, children gain immunity early during infancy. However, the incidence is higher in non-immune hosts. Antibiotic use destabilizes the gut flora and can inhibit the local immune responses, thereby compromising resistance to a variety of infections. It is not yet known whether antibiotic intake can also precipitate C. jejuni enteritis as the infectious dose is low and attack rates are high. We made a preliminary study to determine the prevalence of C. jejuni in hospitalized patients receiving antibiotics for various ailments. One hundred and thirty eight stool samples submitted for Clostridium difficile toxin assay were additionally cultured for C. jejuni in blood-free campylobacter selectivity agar. All suspected colonies were subjected to Gram staining, oxidase, catalase and nalidixic acid sensitivity tests. Confirmation of C. jejuni was done by the hippurate hydrolysis test. Of the 138 faecal samples investigated, 14 (10.1%) grew C. jejuni and 11 of them belonged to adults. Two of these 14 samples were also positive for C. difficile toxin. Though not as yet reported, C. jejuni may also be involved in antibiotic associated diarrhoea due to lowered immunity in the host. It may cause enteritis either by itself or in synergy with C. difficile infection. PMID- 16366119 TI - Beta-lactamase producing Pseudomonas aeruginosa in hospitalised patients. AB - Beta lactamase continues to be the leading cause of resistance to beta lactam antibiotics in gram-negative bacteria. A total of 50 clinical isolates of Pseudomonas aeruginosa were studied to determine the prevalence of ESBL production in hospital strains and also to study their susceptibility to various other antimicrobial agents. ESBL production was observed in a total of 18/50 (36%) of cases. Most of the ESBL positive isolates showed resistance to 3rd generation cephalosporins including multidrug resistance (MDR) to antibiotics like piperacillin, nalidixic acid, ciprofloxacin, levofloxacin, gentamicin and tobramycin. The ESBL producers however showed good susceptibility to drugs like meropenem, gatifloxacin and amikacin. PMID- 16366120 TI - Isolation, distribution and prevalence of various species of enterococci isolated from clinical specimens in a tertiary care hospital. AB - Various clinical specimens were processed to find the prevalence rate of enterococci and to identify the species of clinical isolates of enterococci. Screening of various clinical specimens revealed that enterococci were prevalent in 2% of the total specimens, with urine and wound to be the major site of isolation. Conventional test scheme proposed by Facklam and Collins and commercially available systems Rapid ID 32 Strep (biomereiux) were successfully used to speciate enterococcal strains. Five species of enterococci were identified in the study from a set of 396 cultures, with E. faecalis (79.79%), and E. faecium (11.11%) predominating. E. hirae (3.03%), E. gallinarum (3.03%), and E. casseliflavus (3.03%), were the other members of Enterococcus species identified. PMID- 16366121 TI - Spectrum and antibiotic susceptibility pattern of bacterial isolates from conjunctival swabs. AB - Conjunctivitis is the most common disease of conjunctiva which occurs by exogenous or endogenous source. Bacterial conjunctivitis is the most common type of this infection for which emperical treatment is started without knowing the etiological agents because prior cultures are generally not taken. We conducted this survey to study the etiological agents of conjunctivitis in a total of 6763 swabs obtained over a period of four years (2001 to 2004). The prevalence of bacterial conjunctivitis was found to be 20.4% with a predominance of Staphylococcus aureus (87.2%) followed by Streptococcus pneumoniae (4.7%) and gram negative rods (E.coli + Klebsiella spp. + Pseudomonas spp.) in 8.1% swabs. For S. aureus, amikacin showed maximum sensitivity (73.2%) while for S. pneumoniae, cefotaxime (90.6%) and cefazolin (89%) were found to be highly sensitive drugs. For gram negative rods, ceftazidime (74.1%) and cefotaxime (67%) were found to be preferred options. Thus it is concluded that bacterial conjunctivitis in our hospital is predominated by S. aureus. The bacterial isolates obtained from conjunctiva have not yet attained a high level of antibiotic resistance and hence averts the need to unnecessarily start new/latest generations of antibiotic eyedrops or antibiotic combination which may lead to acquisition of resistance by these bacteria. PMID- 16366122 TI - Prevalence of Trichomonas vaginalis infection in and around Surat. AB - Trichomonas culture method was used for the diagnosis of trichomoniasis along with gram staining and wet mount preparation. We selected 51 urban women and 51 rural women in this study. Samples were taken from both symptomatic and asymptomatic individuals. Out of 102 samples 35 samples were positive for Trichomonas vaginalis by culture method. 20 women were positive for Trichomonas vaginalis by wet mount and 15 by gram staining technique. So it is a fact that culture of Trichomonas vaginalis is a "gold standard" in the diagnosis of trichomoniasis. As we had the opportunity to compare both urban and rural women in our study we were able to come to the conclusion that incidence of Trichomonas vaginalis is more in urban women than in rural women. PMID- 16366123 TI - Application of bleach method in diagnosis of extra-pulmonary tuberculosis. AB - Extra pulmonary tuberculosis (TB) comprises 15% of the total tuberculosis cases. Bleach concentration method for demonstration of Acid fast bacilli (AFB) has been recently described for sputum. The aim of this study is to apply this method for demonstration of AFB in material obtained from extra-pulmonary sites and to correlate with cytology and conventional Ziehl Neelsen (Z N) staining. A total of 55 samples were studied from clinically suspected cases of extrapulmonary TB which included, FNA lymph nodes (17), abscesses drained from various body parts (18), body fluids (18) and skin scrapping (2). All the samples were processed for routine cytology, conventional ZN staining and bleach method followed by ZN staining. Out of 55 samples, 24(43.40%) were indicative of tuberculosis on cytology, 12(21.8%) were positive for AFB on conventional ZN staining, while the positivity increased to 39(70.90%) by Bleach method. Bleach solution is inexpensive and readily available in hospitals and its application has been proved in pulmonary tuberculosis. However to the best of our knowledge this is a pioneer study applied to the extra-pulmonary samples and the results of the present study shows improved detection of AFB. PMID- 16366124 TI - TORCH infection in women with bad obstetric history--a pilot study in Kumaon region. AB - TORCH infections in the mother are transmissible to fetus in the womb or during the birth process and cause a cluster of symptomatic birth defects. In mother they are inapparent or asymptomatic and hence difficult to diagnose clinically. Over a nine months period 20 pregnant women with bad obstetric history were studied. Seropositivity of Toxoplasma, rubella, CMV, and HSV infections (TORCH) were demonstrated by the presence of IgM and IgG antibodies by ELISA method. It was found that, IgM antibodies were positive in 4 cases (20%) for Toxoplasma, 4 cases (28.6%) for rubella and 4 cases (26.7%) for CMV and HSV each. IgG antibodies were positive in 11cases (55%) for Toxoplasma, 10 cases (66.6%) for rubella, 14 cases (93%) for CMV and 11 (73%) for HSV. Therefore all antenatal cases with BOH should be routinely screened for TORCH as early diagnosis and appropriate intervention will help in proper management and fetal outcome. PMID- 16366125 TI - Exercise echocardiography in coronary artery disease. AB - Exercise echocardiography is one of the stress echocardiography modalities, widely used nowadays, mainly in the patients with coronary artery disease. It is a first-line technique for the diagnosis of coronary disease and for prognostication in various patients' subsets, including patients with chronic coronary artery disease, patients with recent revascularization or myocardial infarction and women. This method relies on the fact that in the presence of a significant coronary stenosis, an adequate level of physical exertion produces a mismatch between oxygen demand and supply, resulting in a wall motion abnormality in the area supplied by that stenotic artery. The wall motion abnormalities that develop are detectable by echocardiography and are a marker of the degree of ischemia. The most commonly employed forms of exercise in conjunction with echocardiography are immediate post exercise imaging on a treadmill and exercising during supine or upright bicycle ergometry. Echocardiograms can be interpreted qualitatively, with a description of the recorded wall motion abnormalities, or quantitatively, calculating a wall motion score. The overall accuracy of exercise echocardiography in the diagnosis of coronary artery disease is about 80-90%. Exercise echocardiography has been shown to provide useful information for the purpose of risk stratification and prediction of coronary events after acute myocardial infarction, after angioplasty and after coronary artery bypass grafting. In women, the accuracy of electrocardiographic stress testing is less than in men and stress echocardiography offers a better alternative in the diagnosis of ischemic heart disease. For the patients with an intermediate pre-test probability of having ischemic heart disease and a resting electrocardiogram which precludes an optimal interpretation of the electrocardiographic stress test (digoxin use, left ventricular hypertrophy with repolarization abnormalities, preexcitation, or left bundle-branch block), exercise echocardiography represents a good alternative to radionuclide imaging. PMID- 16366126 TI - Magnesium orotate--experimental and clinical evidence. AB - Magnesium orotate dihydrate (MO) has the sum formula C10H6MgN4O8 x 2H2O and a MG of 370.52. The salt is poorly soluble in water and hence does not bind gastric acid nor does it exhibit noteworthy laxative effects upon oral administration in contrast to easily dissociable Mg salts. As a source of magnesium (Mg), MO is indicated for the oral treatment of extracellular Mg deficiency. Orotic acid (OA), the second active ingredient of MO, is a key intermediate in the biosynthetic pathway of pyrimidines and is shown to improve the energy status of injured myocardium by stimulating, a.o., the synthesis of glycogen and ATP. Myocardial energy-rich phosphate levels are decreased during hypoxic conditions; subsequently, intracellular Mg is depleted and lost via the urine. Since binding sites for Mg (ATP) are provided by OA it can be classified as "Mg-fixing agent". Accordingly MO is also indicated for the treatment of Mg depletion as convincingly shown in animal experiments and also in coronary heart patients undergoing e.g. aortocoronary bypass surgery. PMID- 16366127 TI - Absence of correlation between QRS duration and echographic parameters of ventricular desynchronization. Can we still trust the electrocardiographic criteria? AB - The aim of the study was to assess if echographic ventricular desynchronization parameters measured in patients with dilated cardiomyopathy (DCM) and severe heart failure are correlated with the duration of the QRS on surface electrocardiogram. METHODS: This study included 51 pts aged 58.8 +/- 7.4 years with idiopathic DCM. The following parameters were measured: QRS duration; effective contraction time (ECT) measured as the interval between QRS onset and closure of aortic valve, interventricular delay (IVD) measured as the time between onset of aortic and pulmonary flow, left ventricular mechanical delay (LVD) as the time from maximal interventricular septum contraction and posterior wall contraction, posterior (P), lateral (L) and posterolateral (PL) wall delays, as the time from QRS onset to maximal wall contraction. Regional postsystolic contraction was defined in a given wall as the difference (contraction delay - ECT)> 50ms. RESULTS: 29 pts presented complete LBBB, 22 pts had QRS duration < 120 ms. 39 pts had a postsystolic contraction of the PL wall (32 pts of the L wall and 26 pts of the P wall). 16 pts with QRS duration <120 had a postsystolic contraction of the PL wall (as for the LBBB the rest of 39 pts). In 40 pts the sequence of regional ventricular contraction was: P-L-PL wall (16 pts with QRS<120). LVD was > 100 ms in 36 pts (26 pts with LBBB and 10 with QRS<120). 27 of pts with LBBB and 6 with QRS <120 ms presented IVD> 30ms. There was no correlation between the QRS duration and the parameters listed above. CONCLUSIONS: In DCM pts there is no correlation between the duration of the QRS and echocardiographic parameters of ventricular desynchronization. Mechanical ventricular de-synchrony can be observed in patients with QRS duration < 120 ms. PMID- 16366128 TI - Results of a long-term training program after acute myocardial infarction. AB - BACKGROUND: It is not well known what should be the benefits of a prolonged physical rehabilitation program after an acute myocardial infarction. METHODS: Our study is an open, randomized comparison of a long-term versus a standard rehabilitation program. Sixty-seven patients with acute myocardial infarction were included in an outpatient physical rehabilitation program of 6-8 weeks. Of these, 22 randomly selected patients continued the program until the 36th month (Group A). Twenty-five of the others were rechecked after 36 months, and represented the controls (Group B). For both groups, at the end of this period, a cycloergometer exercise test evaluated the exercise capacity of subjects and an echocardiogram was performed to determine left ventricular systolic and diastolic function. RESULTS: The maximal exercise capacity increased from 147 +/- 13.8 W to 178 +/- 16.4 W in Group A (p < 0.01), but it decreased from 144 +/- 13.2 to 132 +/- 12.8 W in group B. Functional aerobic impairment decreased from 29 +/- 2.7% to 22 +/- 2.1% in Group A, but it increased from 26 +/- 2.5% to 37 +/- 3.8% in Group B. The ejection fraction and diastolic function parameters were not significantly modified during the 36 months, for both groups. CONCLUSION: Long term physical rehabilitation is useful in patients after an acute myocardial infarction to increase effort capacity, but left ventricular performance is not significantly changed. PMID- 16366129 TI - Global myocardial index in patients with multisite pacing. AB - Global myocardial index (GMI) is a sensitive echocardiographic indicator of overall cardiac function. The aim of this study was to compare the GMI changes induced by Bi-V pacing to the variations in clinical, electrocardiographic and echocardiographic parameters currently used for evaluation of ventricular resynchronization in order to test the potential marker of positive clinical response. PATIENTS AND METHODS: This study included 35 consecutive patients (pts), aged 65.7+/-7.5 years, implanted with an endocavitary Bi-V pacemaker for drug refractory severe heart failure. NYHA class was evaluated before implantation and at day 30. QRS duration (QRSd), echocardiographic interventricular delay (IVD), and GMI were determined in the paced and non-paced situation at day 1, 7 and 30 after implantation. RESULTS: NYHA class improved from 3.5+/-0.3 to 2.4+/-0.4 (p<0.01). GMI improved significantly (p<0.0001) after Bi-V in all pts from 1.25 +/- 0.5 to 0.77 +/- 0.34 and remained stable all along the study. GMI decrease (dGMI) in pts with NYHA functional class improvement > 0.5 was 0.52 +/- 0.16while dGMI in pts with NYHA functional class improvement < or = 0.5 was 0.21+/- 0.19. dGMI was statistically correlated with IVD shortening (r=0.67, p<0.0001) but not with QRSd shortening. CONCLUSION: The GMI improves during Bi-V pacing, mostly in responders pts and the improvement remains unchanged after the implant. The GMI decrease induced by Bi-V pacing is correlated with the IVD shortening and NYHA functional class improvement. PMID- 16366130 TI - Streptokinase-induced hypotension has no detrimental effect on patients with thrombolytic treatment for acute myocardial infarction. A substudy of the Romanian Study for Accelerated Streptokinase in Acute Myocardial Infarction (ASK ROMANIA). AB - The Streptokinase (SK) regimen (1.5 MU/60 minutes) has remained unchanged for the past 20 years in patients with ST-segment elevation acute myocardial infarction (STEMI) due to fear of hypotension (a specific effect of this thrombolytic agent) and of hemorrhagic complications. OBJECTIVE: To evaluate the influence of the Streptokinase-induced hypotension (SK-hTA) on the rate of coronary reperfusion (CR), incidence of cardiogenic shock (CS), 30-day mortality and incidence of stroke in patients (pts.) with STEMI. The SK-hTA was defined as decrease of the systolic blood pressure with at least 20% within the first 20 min. after the start of the SK infusion. METHODS: A group of 837 pts. (age 20-90) with thrombolytic treatment, with three "accelerated" SK regimens within the first 6 hours after the onset of STEMI and enrolled in the Romanian open, prospective, non-randomised study for accelerated SK in STEMI (ASK-ROMANIA) have been included. The SK regimens consisted in infusing of the standard dose of 1.5 M.U. either in 30 min. (regimen SK1.5/30, 173 pts).) or in 20 min. (regimen SK1.5/20, 377 pts.) or of the half dose (0.75 M.U.) in 10 min. followed by a new infusion of 0.75 M.U. after 50 min. only if no bed-side signs of CR have been recorded (regimen SK 0.75/10, 287 pts.). The speed of the SK infusion was maintained in all pts. experiencing SK-hTA. All pts. received aspirin and heparin or enoxaparin if not contraindicated. Three noninvasive CR criteria have been used: 1. Rapid cessation of the chest pain. 2. Rapid decrease of the ST segment elevation by more than 50% of the initial value. 3. Rapid increase of the CK and CK-MB with a peak within the first 12 hrs. RESULTS: SK-hTA appeared in 372 pts. (44.55%) at 9+/-5 min after the start of the SK infusion. In this subgroup the rate of CR was 74.46%, non-significantly higher than the one of 68.81% registered in pts. without SK-hTA (p=0.071). SK-hTA disappeared in all patients after 16+/-6 minutes without a specific therapy. Fourteen pts. with SK-hTA (3.76%) and 16 pts. without SK-hTA (3.44%) developed CS after thrombolysis ( non-significant difference). The global in-hospital mortality was 10.21% in pts. with SK-hTA and 9.89% in pts. without this side effect (non-significant difference). The incidences of hemorrhagic and ischemic strokes were 0.26% (1 patient) respectively 0.52% (2 pts.) in the SK-hTA subgroup and 0.43% (2 pts.) respectively 0.64% (3 pts.) in the subgroup without SK-hTA. CONCLUSIONS: 1. Despite a very high incidence (44.55%) the SK-hTA has not a detrimental effect in pts. treated with accelerated SK regimens for STEMI. 2. Streptokinase can be rapidly administered without an increased risk. PMID- 16366131 TI - The tachyarrhythmic cardiomyopathy--a rare entity in childhood. AB - Persistent disturbances of rhythm with high heart rate may lead to reversible depression of left ventricular function and biventricular failure. This phenomenon may appear also in children with incessant supraventricular tachycardia. Because in most cases these patients are long time asymptomatic the diagnosis of these arrhythmias is very difficult and frequently is belated. In case of correct treatment of the arrhythmia, the cardiac function improves rapidly. OBJECTIVE: To present a number of cases of incessant supraventricular tachycardia in children, which has led to tachyarrhythmic cardiomyopathy and heart failure. MATERIAL AND METHOD: A number of six patients (pts.), aged between 3 years 10 m - 16 y, hospitalized between January 1st, 2001 - January 1st , 2004 were taken in study. All patients had long crisis of supraventricular tachycardia with incessant character. A clinical exam, an ECG, a chest X-ray film and an echocardiography were made to all patients. An electrophysiologic study was made in two cases. The emergency therapy was the drug therapy. RESULTS: At the moment of hospitalization all patients had a depression of the contractile function of the myocardium. After the control of the arrhythmia the ejection fraction recovered in a period between 3 months and one year. CONCLUSION: The disturbances of rhythm may induce a depression of the contractile function of the myocardium, which is reversible in many cases. The correct antiarrhythmic treatment led to the improvement of the heart failure in this case. PMID- 16366132 TI - Study of the efficiency of the head-up tilt test with nitroglycerin challenge in the diagnosis of vasovagal syncope. AB - Syncope is defined as a transient, self-limited loss of consciousness. It is an important cause of morbidity in general population and the vasovagal syncope (VVS) is a common clinical problem which often leads to hospital admission, multiple office visits, and performing of many diagnostic tests. Head-up tilt table testing (HUTT) is a widely used diagnostic tool, with proved efficiency in diagnosing the many types of VVS. The aim of our study was to assess the efficacy of the HUTT with sublingual nitroglycerin (NTG) challenge for the diagnosis of vasovagal syncope. PATIENTS AND METHODS: The study groups consisted of 72 patients (37 women, 35 men), mean age (+/-SD) 38.5 +/- 15.7 years, referred to our clinic for syncope of unknown origin and 16 healthy volunteers as control subjects (9 women and 7 men) with a mean age 26 +/- 6.5 years and no history of syncope. The HUTT protocol was performed in the morning after an overnight fast, with a 15 minutes supine equilibration phase followed by a 30 minutes drug free tilt phase at an angle of 70 degrees. If no positive response was recorded, 400 micrograms of NTG spray were given sublingually and the HUTT continued for 20 minutes. RESULTS: The HUTT was positive in 58 patients (80.5%) and negative in 14 (19.5%) while in the control subjects 4 (25%) had positive HUTT response. The sensitivity of the HUTT protocol with nitroglycerin challenge used in our study was 81% and the specificity was 75% (p<0.0001). CONCLUSION: The described HUTT protocol appears to be a simple and efficient tool for the diagnosis of syncope associated with normal ECG and no signs of organic heart disease. The sublingual nitroglycerin challenge was safe to use and showed no side effects while keeping the sensitivity and specificity of the test to an accepted level. PMID- 16366133 TI - Amiodarone-induced thyroid dysfunction in cardiac patients from areas with iodine deficiency. AB - Amiodarone via iodine excess can determine thyroid dysfunction. AIM: to assess thyroid dysfunction in patients treated with amiodarone, according to previous daily iodine intake. MATERIAL AND METHODS: 63 patients treated with amiodarone were assessed. 11 of 63 were resident in a moderate iodine deficient area. Thyroid stimulating hormone (TSH), free and total thyroxine (T4) and total triiodothyronine (T3) were measured. Thyroid ultrasonography, color flow Doppler sonography (CFDS), radioiodine uptake (RAIU) at 2 and 24 hours were also performed. RESULTS: Amiodarone-induced thyrotoxicosis developed in 31 patients (49.2%); 17 patients (27%) remained euthyroid. Patients from iodine deficient areas developed more frequent hyperthyroidism (91% vs. 40.4%), at significant lower cumulative doses of amiodarone, and never hypothyroidism. Overt hyperthyroidism prevails (29/31 patients). Frequency of amiodarone-induced thyrotoxicosis type I was 19% (12/63), type II 12.7% (8/63), and of mixed forms 17.5% (11/63), without significant differences between the two geographical areas. There are no peculiar features of amiodarone-induced thyrotoxicosis versus common hyperthyroidism, but there is a tendency at recurrence of tachyarrhythmias at lower levels of T3 than in common hyperthyroidism. Amiodarone-induced hypothyroidism developed in 15 patients (23.8%) and none was from iodine deficient areas, but almost half show high levels of antithyroid peroxidase antibodies (ATPO). Subclinical hypothyroidism prevails (11/15 patients). CONCLUSION: a predictor for amiodarone-induced thyroid dysfunction is iodine deficiency in nutrition. PMID- 16366134 TI - Epidemiology of acute myocardial infarction in Romanian county hospitals: a population-based study in the Baia Mare district. AB - The epidemiological characteristics, the methods used in treating the patients, as well as the evolution of the myocardial infarction are not known in the Romanian county and municipal hospitals. MATERIAL AND METHODS: Six hundred and fifty one cases of acute myocardial infarction were retrospectively recorded in the Emergency County Hospital of Baia Mare from 1.01.1999 till 30.09.2003. Data on treatment delay, therapeutic strategies, duration of hospitalization and outcome were collected. The study was completed by a postal investigation which was carried out from the 1st of October until the 31st of December 2003. This investigation consisted in one application form (multiple choice answers were requested) which had been handed out to the patients who survived the acute episode and left the hospital. RESULTS: The first medical care took place for 60.9% of the patients in hospital. About 72% of these patients were male and the mean age was 66 +/- 12 years. The types of myocardial infarction with ST-segment elevation STE-MI represented 67%, followed by 29.5% with no ST-segment elevation NSTE-MI, and 3.5% with undetermined ECG. The maximum values of Killip class during the first 48 hours were: I in 55% of the patients, II in 23%, III in 14% and IV in 9% of them. The thrombolytic therapy was done in 31% of the patients who were recommended to receive a reperfusion treatment. During their first period in hospital, 37% of the patients had an effort test and 2.45% were transferred to a tertiary care hospital for invasive procedures. The overall median hospital stay was 9 days. The mortality in hospital was 13.6%, respectively 8.1% in the study group who received the reperfusion therapy and 15.2% among those who had followed the standard care. The results of the mail survey revealed that only 62% of the patients who answered the questionnaire (N=498) had regular cardiologic check ups and 43% underwent a treatment with beta blockers and 12% a treatment with statins. Out of these patients 16% had a revascularization procedure that was performed in a specialised University Hospital. The overall mortality in the responder group was 35.5% with an estimate 7.5% year rate. CONCLUSIONS: The analysis of the present data and especially the rate of mortality suggest until two times higher values in comparison to the European average. These results demonstrate the fact that it is imperative to generalize the thrombolytic therapy in all the hospitals in Romania and give a wider access to the myocardial revascularization therapies. PMID- 16366135 TI - Recurrent atrial myxoma. AB - We have chosen this case of sporadic atrial myxoma for our presentation because it had a particular evolution, with recurrence at 8 years after surgical excision (echocardiography was performed every year) and a particular diagnostic means - at echocardiographic follow-up, the patient being asymptomatic. This presentation, together with a review of literature included in the article, emphasizes the importance of a careful postoperative follow-up of the patients and the existence of some particular aspects of the evolution and symptomatology of recurrent atrial myxoma. PMID- 16366136 TI - New insights into the pathogenesis and prognosis of aortic sclerosis. AB - Aortic valve disease includes a wide spectrum of conditions, from leaflets thickening to hemodynamically significant aortic stenosis and is one of the most important causes of morbidity and mortality worldwide. Studies performed during the last years bring consistent evidence that these lesions of the aortic valves share remarkable similarities with atherosclerotic lesions. They also reveal new risk factors for its development, which are superposed in fact with the major risk factors for atherosclerosis. Moreover, large prospective studies reported an increased risk for cardiovascular events and for total and cardiovascular mortality in patients with aortic sclerosis. Hence, recent papers also propose therapeutic targets, using statins for delaying the progression of the disease. PMID- 16366137 TI - Microvolt T wave alternans (MTWA)--a new non-invasive predictor of sudden cardiac death. AB - Over 10 million people are at high risk of sudden cardiac death (SCD). In order to stratify the risk of SCD due to ventricular tachyarrhythmia, additional methods are required. In the past years, the development of implantable cardioverter/defibrillator (ICD) has become an effective therapeutic option for patients at high risk for SCD, albeit ICD is both expensive and invasive. The Microvolt T-Wave Alternans Test (MTWA, Cambridge Heart Inc.) is a non-invasive method with a stronger positive predictive value in the assessment of arrhythmic risk, when compared to invasive electrophysiological studies (EPS). MTWA has been proved at least an equivalent or even a more sensitive risk predictor of ventricular tachyarrhythmia and/or SCD. MTWA may be considered as a first-line screening test for patients with ventricular arrhythmia history, in order to discriminate the high risk / low risk subgroups, prior to invasive EPS and/or treatment. PMID- 16366138 TI - Cardiovascular risk and rheumatoid arthritis: from mechanisms of atherosclerosis to therapeutic approach. AB - Large epidemiological studies showed that one of the most important causes of death in patients with rheumatoid arthritis (RA) is represented by cardiovascular disease. Thus, the presence of RA is associated with an increased risk of the occurrence of stable angina, myocardial infarction, heart failure and stroke. However, studies performed during the last years failed to bring us clear evidence regarding the role of traditional cardiovascular risk factors (hyperlipidemia, diabetes mellitus, hypertension, smoking and obesity) in the pathogenesis of cardiovascular disease in these patients. Recently, the role of inflammation and its mediators not only in the atherosclerosis plaque development but also in the mechanisms of vulnerable plaque was clearly demonstrated. From this point of view, recent studies showed that inflammatory cells and mediators of inflammation are both markers of an increased cardiovascular risk and unfavorable cardiovascular outcome, and also cardiovascular risk factors that act in an active manner in the processes that promote atherosclerosis. Taking into account the fact that RA is a systemic inflammatory status, recent reports demonstrated the involvement of inflammation mediators in connection with prothrombotic factors and endothelial dysfunction in the development of cardiovascular disease in RA patients. There are only scarce data in the literature regarding the benefice of cardiovascular risk reduction therapies in this group. Further studies are required for the refinement of the cardiovascular risk stratification algorithms and for the improvement of the cardiovascular risk management in rheumatoid arthritis. PMID- 16366139 TI - Chronic inflammatory rheumatismal diseases--paraclinical diagnosis. AB - The rheumatismal diseases constitute a major problem of public health, the pathology of the locomotor system representing today the main cause of invalidity in the world. The inflammatory chronic rheumatisms affect the young population, and, between the onset of the disease and diagnosis (approximately one year), the patient does not accuse major clinical complaints. In this context, the present work proposes a presentation of the main paraclinic methods and techniques, which may direct the clinician in giving a correct and early diagnosis of the main chronic inflammatory rheumatismal disorders. The paraclinic methods and techniques for assessing the intensity and evolution of the chronic rheumatismal inflammatory processes may be classified as follows: genetic, biological, histological, and imagistic. The inflammatory reaction is a nonspecific long-term reaction of the body defense systems, as response to the continuous and recurrent aggression. It is characterized by the predominance of the lymphocytes and macrophages at the level of the focus infectious and an intense process of neoangiogenesis and fibroblastic proliferation accompanies it. The paraclinic methods and techniques for assessing the intensity and evolution of the chronic rheumatismal inflammatory processes may be classified as follows: genetic, biological, histological, and imagistic. PMID- 16366140 TI - Ankylosing spondylitis--a systemic disease. AB - Ankylosing spondylitis (AS), the prototype of seronegative spondylarthropathies, is in fact a systemic disease, causing numerous extraskeletal manifestations that have a significant influence on patients vital prognostic. This article presents the principal systemic manifestations of AS, pointing out pathogenetic, clinical and therapeutic aspects. PMID- 16366141 TI - A Romanian instrument to facilitate bone density measurement indication in postmenopausal women. AB - The aim of the study is to find and valid a clinical instrument which identifies the women that need a bone density measurement because of their high risk for osteoporosis. MATERIAL AND METHOD: A number of 356 women were enrolled in this study. They filled in a study formular and their bone density was measured by DEXA exam (dual energy X-ray absorbtiometry) in the lumbar spine and proximal femur. Statistical correlations between the dependent variable minimal T-score and the other variables (clinical risk factors) were established. The most significant osteoporosis risk factor were included in a classification tree for the dependent variable minimal T-score with 5 splits, 5 independent splitting variables and 6 terminal knots. RESULTS: The most significant osteoporosis predictors included in our classification tree are: current weight, decreasing of height, sedentary life, alcohol consumption, number of deliveries. In predicting osteoporosis, the classification tree has high values both for sensitivity and for specificity (88.2%, respectively 74.11%).The osteoporosis prevalence is very high (74.34%) within the group predicted as having osteoporosis (53.7% of all women). The osteoporosis prevalence was 11% within the group predicted as having osteopenia. CONCLUSIONS: The classification tree facilitates a bone density measurement strategy and establishes the criteria for initiation of the treatment in high risk osteoporosis cases. It allows postponing the DEXA-exam for low risk women and selective measurement of the bone density for intermediar osteoporosis risk women. PMID- 16366142 TI - Clinical relevance of antikeratin antibodies in rheumatoid arthritis and symmetric polyarthritis associated with hepatitis C infection. AB - Chronic hepatitis C virus (HCV) has been linked to extrahepatic autoimmune phenomena. In addition a variety of autoantibodies were found in patients with HCV. This study was performed to assesss the clinical relevance of antikeratin antibodies in rheumatoid arthritis (RA) and in patients with symmetric polyarthritis associated with hepatitis C infection. Serum antikeratin antibodies were evaluated in 3 different groups of patients; all were rheumatoid factor (RF) seropositive: Group 1: 31 patients with HCV associated symmetric polyarthralgia or arthritis. Group 2: 28 patients with RA (modified ACR criteria for probable RA). Group 3: 16 patients with autoimmune disorders other than RA. Seventeen healthy individuals matched for age and sex served as controls. In our study, 75 patients who were rheumatoid factor positive (measured by ELISA, the cutoff was established to 20 U/mL) were tested for antikeratin antibodies using an indirect immunofluorescence technique with 1:10 serum dilution. Antikeratin antibodies were detected in 18/28 (64%) patients with true RA and only 3/31 (9%) patients with HCV-related arthritis (p < 0.0001). Antikeratin antibodies were observed in 3/16 (18%) patients of group 3 (p < 0.05). Antikeratin antibodies were not found in the sera of the healthy controls. PMID- 16366143 TI - Association between primary Sjogren's syndrome and non-Hodgkin's lymphoma. AB - Sjogren's Syndrome (SS) is one of the most frequent autoimmune disorders, which affects approximately 1% of the population. It occurs in patients of all ages, but especially females during the fourth and fifth decades of life with a female/male ratio of 9:1. The main target of this disease are the exocrine glands that are infiltrated progressively by lymphocytes and finally destroyed, leading to decreased exocrine secretion. Thus primary SS is usually defined as xerophtalmia (dry eye) and xerostomia (dry mouth) accompanied in 60% of cases by parotid swelling [1]. The most serious complication of Sjogren's Syndrome is the high risk of the occurrence of non-Hodgkin's Lymphoma. PMID- 16366144 TI - Lupus-like disease with anticardiolipin antibodies and lupus anticoagulant as a cause for atypical Eaton-Lambert syndrome and peripheral nerve disease: a case report. AB - In the practice of neurology, the type of clinical involvement suggests the site of the lesion and the mechanism beneath. Sometimes, the symptoms can be delusive, turning the diagnostic approach to a wrong path and raising the necessity of an algorithm considering the less probable entities. Dysimmunity as mechanism of neurological disease involving both the neuromuscular junction and peripheral nerves is frequently insidious and difficult to suspect on clinical basis alone. We report the case of a 67-year-old male with atypical Eaton-Lambert syndrome and mononeuropathy probably in relation with lupus-like entity. The patient has also high titers of anticardiolipin antibodies and lupus anticoagulant. We are considering the diagnostic algorithm before an isolated and atypical neurological presentation and reviewing the main neurological manifestations in lupus-like and autoimmune systemic disease. We raise the difficulty to classify an inflammatory connective tissue disease in the absence of other pathologic features than autoantibodies and isolated neurological symptoms and discussing the main therapeutic issues. PMID- 16366145 TI - [Electroencephalographic correlates of brain states during verbal learning: Communication II. Characteristics of spatial synchronization of EEG]. PMID- 16366146 TI - [Psychophysiological parameters of the functional state in adolescents at different stages of puberty during intense informational loading]. PMID- 16366147 TI - [Characteristics of the theta and alpha EEG bands in adolescents of 15-17 years old]. PMID- 16366148 TI - Heart rate differences between targets and non targets in intuition tasks. AB - This study reports the results of one experiment and a replication, aimed at investigating heart rate changes related to a pure intuition task. In each experiment, twelve subjects were required to guess which of the four pictures presented in sequence for about 10 seconds, was the target. Each subject performed 20 trials. In each trial the target was automatically selected using a pseudo-random algorithm. Heart rate was recorded (see Method section for details) during the pictures presentation. In the first experiment, a statistical significant increment of heart rate associated to targets with respect non targets was observed. The replication experiment with new twelve subjects confirmed the data obtained in the main experiment. These findings support the hypothesis that heart rate is related not only to overt but also to covert cognitive activity such as that involved in intuition tasks, giving convergent evidence to the models describing our intuitive cognitive activity as a double, partial independent information processing system. PMID- 16366149 TI - [Assessment of heart autonomic control on the basis of spectral analysis of heart rate variability]. PMID- 16366150 TI - [Voluntary control of thoracic and abdominal respiratory movements]. PMID- 16366151 TI - [Seasonal parameters in newborns under conditions of the European north]. PMID- 16366152 TI - [Heart rate variability in subjects with increased motor activity and athletes]. PMID- 16366153 TI - [Electrocardiographic manifestations of chronic physical overexertion in athletes]. PMID- 16366154 TI - [Comparative analysis of the results of a stepped running test performed in the vertical and horizontal positions]. PMID- 16366155 TI - [Biochemical markers of bone tissue metabolism in cosmonauts after a prolonged spaceflight]. PMID- 16366156 TI - [Informative value of temperature parameters in different areas of the human body for correcting body thermal imbalance during extravehicular activities in cosmos]. PMID- 16366157 TI - [On the possibility of correction of the peripheral nervous system pathology by physical factors emerging during the functioning of EPR, NMR, and NGR]. PMID- 16366158 TI - [Intestinal endotoxin in regulation of hemostasis activity and in pathogenesis of the DIC syndrome]. PMID- 16366159 TI - Cardioprotection by aldosterone receptor antagonism in heart failure. Part I. The role of aldosterone in heart failure. AB - In recent years understanding of the role of aldosterone has expanded beyond the known classic effects of promoting renal sodium retention and potassium and magnesium loss. It is now well documented that aldosterone causes myocardial and perivascular fibrosis, blocks the myocardial uptake of norepinephrine, and increases plasminogen activator inhibitor levels. In conjunction with angiotensin II, aldosterone causes vascular damage, endothelial dysfunction, and decreased vascular compliance. Therefore, the renin-angiotensin-aldosterone system (RAAS) plays a major role in the development of both hypertension and heart failure and is therefore, a key target for therapeutic interventions. Commonly prescribed medications for control of hypertension and congestive heart failure are inhibitors of the RAAS, including angiotensin converting enzyme inhibitors (ACE I) and Angiotensin II (A-II) receptor antagonists. There is a well-documented increase in aldosterone levels that occurs over several months during chronic treatment with an ACE-I or A-II receptor antagonist. Such suppression of circulating aldosterone however, is transient, as exemplified by the term "escape" used to describe the phenomenon. This rebound of aldosterone even occurs when patients receive both an ACE-I and A-II receptor antagonist. In addition, ACE-I and A-II receptor antagonists are less effective in controlling BP in the estimated 60% of hypertensive patients who are salt (volume) sensitive and more prone to hypertension-associated morbidity such as black patients and type 2 diabetics. Thus chronic and complete blockade of aldosterone action requires an aldosterone receptor antagonist. The "Randomized Aldactone Evaluation Study" (RALES) trial results in patients with severe heart failure NYHA class III or IV and a left ventricular ejection fraction of no more than 35 percent showed that administration of a sub-hemodynamic dose of spironolactone (25 mg a day) as an add on therapy to ACE-I plus standard treatment resulted in a significant mortality reduction due both to decreased death from progressive heart failure and sudden cardiac death. These findings support the pivotal role of aldosterone in the pathophysiology of progressive heart failure. Although it is an effective antialdosterone agent, widespread use of spironolactone in humans is limited by its tendency to produce undesirable sexual side effects. At standard doses, impotence and gynaecomastia can be induced in men, whereas pre-menopausal women may experience menstrual disturbances. Data on a selective aldosterone receptor antagonist, eplerenone, appear promising for the effective blockade of aldosterone and its harmful effects without the sexual disturbances of spironolactone. Recently Eplerenone was successfully introduced for the treatment of hypertension and heart failure. Growing number of experimental studies are finding a broader role for Aldosterone in driving the pathophysiology of both heart failure and hypertension. When added to conventional therapy aldosterone receptor blockers show benefits which are in addition to those conferred by ACE-I and/or AII receptor blockers. PMID- 16366160 TI - [Problems of studying executive function, regulation, and control of mental activity in humans]. PMID- 16366161 TI - [Method for investigating the patellar reflex under isometric conditions]. PMID- 16366162 TI - [Parameters of EEG in children of early preschool age with consequences of perinatal lesions of the CNS]. PMID- 16366163 TI - [Changes in the spatial-contrast sensitivity during work with liquid-crystal monitors and with spectral correction of vision]. PMID- 16366164 TI - [Gender differences in subjective time perception and sensitivity to antianxiety drugs]. PMID- 16366165 TI - [Usefulness of rational approaches in cardiology]. PMID- 16366166 TI - [Modification of a hydraulic device for controlled banding of the trunk of the pulmonary artery in dogs]. AB - Different congenital or acquired diseases increase the right ventricular systolic pressure. The elevation is achieved by pulmonary artery banding with linen tape or hydraulic devices. We introduce a new hydraulic device to be used in experimental subjects of less than 15 kilograms. METHODS: An experimental laboratory phase was conducted. In this phase, the hydraulic devices were tested against pressures as high as 80 mm Hg; later on the hydraulic devices were placed on the dogs' main pulmonary artery; the right ventricular systolic pressure, pulmonary artery pressure, and their differences were obtained. The values are expressed in average +/- standard deviation. RESULTS: The average weight of the dogs was 12.6 +/- 2.2 kg. The average diameter found in the main pulmonary artery was 12.4 +/- 1.4 mm and 27.5 +/- 1.6 mm length. Right ventricular systolic pressure of basal condition to maximum pressure obtained was 25.5 +/- 1.9 mm Hg vs 50.3 +/- 6.9 mm Hg, p < 0.0001; pulmonary artery pressures under the same conditions were 24.7 +/- 2 mm Hg vs 21.5 +/- 6.9 mm Hg, p < 0.043; and the differences between them were 0.8 +/- 1.4 mm Hg vs 28.8 +/- 4.2 mm Hg, p < 0.0001, developed by the hydraulic device respectively. CONCLUSIONS: The hydraulic device allows attaining a controlled acute increase in right ventricular systolic pressure. PMID- 16366167 TI - [Myocardial regeneration in Ambystoma mexicanum after surgical injury]. AB - Ventricular resection of the heart of Ambystoma mexicanum was performed and the type of tissue that restored the lesion and if it is by hypertrophy or hyperplasia of myocardium, were evaluated. Masson's trichrome stain indicated that 5 days after resection, the gap was occupied with a blood clot surrounded by collagen fibres (83 +/- 6%) and muscle (10 +/- 3%) and the rest of area (7 +/- 2%) free of tissue. A proportion of 50 +/- 4 and 90 +/- 2% was muscular tissue, 10 and 30 days after injury. The evaluation with bis-Benzimide indicated cell proliferation in the injured area. The double immunohistochemistry for alpha sarcomeric actin and proliferating cell nuclear antigen indicated that the tissue that occupied the injury-produced gap was originated by cardiomyocyte proliferation, which presented a maximum of 68%, 5 day after injury. Our results indicate that the myocardium of A. mexicanum recovers its structure through cardiomyocyte hyperplasia and suggest that the myocardial regenerative capacity is higher than the reported for adult mammals (1%) and other non-mammalian vertebrates (32%). This characteristic makes A. mexicanum a suitable model to study the mechanisms that regulate per se, the myocardial regeneration in adult vertebrates in vivo. PMID- 16366168 TI - [Prothrombotic state in early stages of chronic Chagas' disease. Its association with thrombotic risk factors]. AB - OBJECTIVE: The aim of this study was to explore the presence of prothrombotic state in early stages of chronic Chagas' disease with serum markers of thrombosis and fibrinolysis, and to investigate it's association with thrombotic risk factors for venous thromboembolic disease. PATIENTS AND METHODS: Forty two patients with chronic Chagas' disease were compared with 21 healthy volunteers. Thrombotic markers used were fragment 1 + 2, ATM complex, fibrinogen/fibrin degradation products, D-dimer and beta-thromboglobulin. Fibrinolysis was evaluated with euglobulin lysis time, tissue plasminogen activator and it's inhibitor levels. A thrombophilic screening was performed. Antithrombin and protein C were determined by functional methods, as well as free fraction of protein S, resistance to activated protein C, factor V Leiden R506Q mutation, prothrombin G20210A mutation, homocysteine and antiphospholipid antibodies: lupus and anticardiolipin antibodies isoforms IgG and IgM. RESULTS: In chronic Chagas' disease patients, statistically significant differences were observed in thrombotic markers: fragment 1 + 2 (p < 0.0001), ATM complex (p < 0.0001), fibrinogen/fibrin degradation products (p < 0.05) and D-dimer (p < 0.05). beta thromboglobulin did not reach statistically significant difference (p = 0.06). Statistically significant differences (p < 0.0001) were found only in euglobulin lysis time, a non specific fibrinolytic marker. Specific fibrinolytic markers tissue plasminogen activator and it's inhibitor, however, did not show statistically significant differences among studied groups. CONCLUSIONS: Eighty six percent of patients had positive thrombophilic screening for at least one thrombophilic risk factor. Thrombophilic risk factors were inherited in 39% and acquired in 83% of the patients. PMID- 16366169 TI - [Antibodies against T. cruzi in patients with dilated cardiomyopathy in Tuxtla Gutierrez, Chiapas]. AB - Chagas disease is caused by the flagellate protozoan T: cruzi. Seroepidemiological surveys in Chiapas, Mexico have shown seropositive individuals, therefore, we searched for people affected by the chronic form of Chagas disease which involves the heart, causing a chronic, progressive and fatal disease called Chronic Chagasic Cardiopathy (CCC). MATERIAL AND METHODS: To establish the frequency of CCC we studied 28 patients seen at the Hospital General Regional "Dr. Rafael Pascacio Gamboa" during October 2002 through October 2003 in Tuxtla Gutierrez, Chiapas, the State capital city, with diagnosis of dilated cardiomyopathy (DC), a serological survey for antibodies against T. cruzi was done. This hospital cares for people from all parts of Chiapas, Mexico. Clinical diagnosis of DC was established there and blind serological studies were performed in Mexico City. RESULTS: Fifteen out of 28 DC patients (54%) had anti T. cruzi antibodies. All of them came from poor rural villages and they had heart failure and/or arrhythmia or heart blockade on EKG. CONCLUSIONS: This observation suggest that in Chiapas were Chagas disease is endemic, there are CCC patients. Any case with a clinical diagnosis of DC should be tested for antibodies against T. cruzi. The low socioeconomic status, culture and environment in this Mexican State favour the presence and transmission of this parasitic disease. PMID- 16366170 TI - [Levosimendan in patients with acute cardiac failure]. AB - OBJECTIVE: To assess levosimendan efficacy in acute cardiac failure. METHODS: We included 25 patients with acute cardiac failure and partial conventional therapy response defined as persistence of low cardiac output with inotropic support. We started levosimendan at loading dose and continuous infusion for 24 hours, recording hemodynamic data, as well as clinical variables. RESULTS: All patients showed clinical and hemodynamic improvement. Pulmonary wedge capillary pressure (PWCP) decreased from 18 cmH2O (11-30 cmH2O) to 14 cmH2O (8-15 cmH2O) at 12 hours (p = 0.3) and to 13 cmH2O (8-12 cmH2O) at the end of the infusion period (p = 0.2). Cardiac index increased from 2.1 L/min/m2 (1.8-3.5 L/ min/m2) to 2.86 L/min/m2 (2-4.3 L/min/m2) at 12 hours (p = 0.03) and to 3.08 L/min/m2 at the end of the infusion period (p = 0.02). Tachycardia higher than 120 bpm was present in six patients. No major hypotensive events were present. Mortality to the present is of eight patients. CONCLUSION: Levosimendan infusion improved hemodynamics and clinical variables in acute cardiac failure with few side effects in this small series of patients. PMID- 16366171 TI - [Leucocytosis associated with higher incidence of adverse cardiovascular events in myocardial infarcts]. AB - In acute phase leukocytes has been related with higher incidence of major adverse cardiovascular events, (MACE) this evidence in AMI ST elevation after mechanical reperfusion is poor. We study prospectively this group to relationship among mechanical reperfusion, leukocytes and MACE. Two groups were considered < 10,000/microL or > 10,000/microL; 271 patients had full inclusion criteria in a 5 year period; 93 had < 1 0,000/microL leukocytes. (8,300 +/- 1,254/microL) and 178 > 10,000/microL. (13,810 +/- 3,192/microL, p 0.0001). We did not observe any difference between both groups regarding demographic characteristics. At beginning leukocytosis group had higher flow TIMI 0 - 1 incidence (89% vs 75%, p 0.004) and in-hospital major cardiovascular adverse events (32% vs 14%, p 0.001) and in follow-up (5% vs 2%, p 0.04). Logistic regression model include > 60 years old, diabetes, extensive anterior or inferior infarction, TIMI flow 0, 1, or 2, cardiogenic shock, leukocytosis and neuthrophilia, had close relationship with mortality (p = 0.0007, RM 1.40, IC 95% 0.410 - 4.841). Multiple regression that include leukocytosis and neuthrophlia had stronger correlation with major cardiovascular adverse events (mortality, r = 0.34 and cardiogenic shock, r = 0.27) and abnormal TIMI flow (r = 0.20). CONCLUSION: Our results confirm close relationship among leukocitosis, thrombosis and major cardiovascular adverse events and extend this knowledge to acute phase and follow- up in acute myocardial infarction ST elevation under percutaneous coronary intervention. These results could be considered as evidence that connecting between endotelial dysfunction (inflammation-atherothrombosis) and cardiovascular disease. PMID- 16366172 TI - [Double sequential electrical transthoracic shocks for refractory atrial fibrillation]. AB - BACKGROUND: Clinical studies have shown that transthoracic cardioversion of atrial fibrillation is dependent on achieving adequate current flow to the heart, which is dependent on transthoracic impedance. When multiple standard cardioversion fails to restore sinus rhythm in patients with atrial fibrillation the double sequential transthoracic shock may be an alternative. METHODS AND RESULTS: Twenty one consecutive patients with paroxysmal or persistent atrial fibrillation refractory to at least two initial high energy 360 J or 200-300 and 360 J monophasic shocks underwent double sequential shocks with 720 J by means two defibrillators. Mean age was 64 +/- 11 years and mean weight 97 +/- 19 kg (range, 49 to 112). Duration of atrial fibrillation was present < or = 3 months in 76%. Arterial hypertension was present in 38% and lone atrial fibrillation in 33%. Mean left atrial size was 4.5 +/- 0.7 cm (range, 3.5 to 6.0). Sinus rhythm was achieved in 19 (90.4%). Two refractory to biphasic shocks with a median 1,050 J (range, 660 to 1,440 J) without major complications. Multivariate analysis identified duration of atrial fibrillation, > 90 days (RR 0.96, CI 0.95-0.98 p = 0.02) and body weight, 101 +/- 11 kg (RR 0.64, CI 0.46-0.90 p = 0.01) variables independently associated with cardioversion unsuccessful. Patient weight, p = 0.002 was the univariate predictor of unsuccessful cardioversion. High energy cardioversion does not cause cardiac damage evidenced from cardiac troponin T estimation. CONCLUSION: For refractory atrial fibrillation to conventional cardioversion double sequential transthoracic shocks represents a safe and highly efficacious alternative and may have a general applicability. PMID- 16366173 TI - [Association of biventricular resynchronizer and implantable cardioverter defibrillator]. AB - Congestive heart failure (HF) remains a major and growing public health problem despite recent therapeutical developments. Thirty to sixty percent of patients with dilated cardiomyopathy (DCM) die suddenly from cardiac arrhythmias. Cardiac resynchronization therapy (CRT) and implantable cardioverter defibrillator (ICD) therapy are effective treatments for HF with a wide QRS and for ventricular arrhythmias respectively. Several trials are currently being performer to evaluate the cardiac resynchronization and implantable cardioverter defibrillator therapy with good results. The objective of this paper is to report the first three patients, in Mexico, that have received this combined therapy. In one patient, a three cameral pacemaker was associated with a unicameral ICD and the other two received a device with both functions. Patients were men, aged 63, 65, and 54 years, two of them with previous myocardial infarct and functional class II to IV of the NYHA. Left ventricular ejection fraction was of 25% in two patients and of 35% in the other. All patients improved their functional class and LVFE, two patients presented discharges of the ICD. PMID- 16366174 TI - [Hemorrhagic pericarditis and cardiac tamponade in systemic lupus erythematosus. A case report]. AB - Systemic lupus erythematosus is a chronic inflammatory autoimmune disorder that can affect any organ or system. Although pericarditis is the most frequent cardiac manifestation of this entity, usually is not a life threatening situation. Pericardial effusion causing cardiac tamponade is a very rare complication in lupus, with an incidence less than 2%. We report a case of pericardial tamponade due to SLE with severe hemodynamic involvement in a 21-year old woman associated to rapidly progressive glomerulonephritis, acute pancreatitis, acute acalculous cholecystitis, pleural effusion, hematologic, cutaneous and neurologic lupus activity. Recognition of this rare manifestation of SLE may be life saving. PMID- 16366175 TI - [Pacemaker lead implant via the persistent left superior vena cava]. AB - BACKGROUND: Persistent left superior vena cava (PLSVC) is a structural, asymptomatic and infrequent anomaly, present in 0.5% of the general population. Typically the diagnosis reveals itself unexpectedly at the time of pacemaker implantation, when it acquires anatomic relevancy. Several techniques are used for the transvenous approach to enter the central venous circulation; the left subclavian vein has become a common access site for electrode implantation and, occasionally, one can find an anomalous venous structure such as a PLSVC. Placement of electrodes through this anomalous venous structure can prove challenging, if not impossible. The present report tries to explore aspects of transimplantation diagnosis from a practical point of view. It also address the knowledge of anatomy, implant technique and radiographic orientation. CASE PRESENTATION: Twenty-six-year-old woman with confirmed Symptomatic Sick Sinus Syndrome variety Sinus Arrest. The diagnosis of PLSVC was discovered unexpectedly at the time of the transvenous approach. The tip for the diagnosis was the "unusually medial" position of the lead, and the venogram showed the venous traject towards the coronary sinus and drainage into the right atrium. An active fixation screw-in electrode was positioned in the antero-superior margin of the free wall of the right atrium. After 24 months of successful placement of the pacemaker, the patient is asymptomatic. DISCUSSION: PLSVC is a rare congenital vascular abnormality. Besides its association with congenital anomalies, its most relevant clinical implication is the association with disturbances of cardiac rhythm, impulse formation and conduction. The ontogenetic development of the sinus node, the atrioventricular node, and the His bundle might be heavily influenced by the lack of regression of the left cardinal vein. When isolated, the PLSVC is usually not recognized until left superior approach to the heart is required, when it becomes a relevant anatomic finding. In fact, it can complicate the positioning of left-sided pacemaker and cardioverter-defibrillator leads. In patients with poor handling through the coronary sinus, a right approach is recommended after visualization of a right superior vena cava entering the right atrium by echocardiography or contrast venography since its absence or hypoplasia (which is reported in 10% of the cases with PLSVC) may represent a major obstacle and would suggest an epicardial implantation. CONCLUSION: Today, the preferred approach for pacemaker lead implantation is via the left subclavian vein and the operator must be aware of this venous anomaly that may technically complicate the electrode positioning. This kowledge may be useful for other medical specialties that require the implant of left sided transvenous subclavian catheters, like in critical care settings, nephrology, onco-hematology, and anesthesiology. PMID- 16366176 TI - [Cardiac rhabdomyoma surgically treated with success. Review of literature]. AB - The primary cardiac tumors are inusual, the incidence varies in all the ages between 0.005 to 0.05%. In pediatrics patients the incidence is 0.27%. The more frequent tumors during the childhood are the cardiac rhabdomyomas. These tumors are considered benigns. The clinical expression is wide, in the most the cases, the patients are asymptomatic and are detected by murmurs. In the prenatal age are manifested by arrhythmias or hydrops fetalis. The neonates and children may be show cardiac arrhythmias, low cardiac index and sudden cardiac death. The association with tuberous sclerosis had been reported in 81%. We present a neonate with cardiac rhabdomyoma diagnosed in the newborn period when he was asymptomatic, however in the follow-up he developed cardiac failure by obstruction in the out flow tract of the right ventricle. He underwent open cardiac surgery to resect the obstruction. Five months after surgery, the patient remain asymptomatic. PMID- 16366177 TI - [The centennial of blood coagulation doctrine]. AB - For centuries, the mystery surrounding blood coagulation stimulated the curiosity of researchers. The knowledge about this function has increased notably in the last century and has permitted to understand numerous physiopatho logical phenomena in several hemorrhagic and thrombotic diseases, and has made it possible to develop diverse drugs of proved efficacy for prevention and therapy. All this was initiated in 1905, when Paul Morawitz published an extensive monograph on the four factors of blood coagulation known until then (fibrinogen, thrombin, thrombokinase, and calcium). In that work, he proposed a blood coagulation model in two stages: thrombin generation and fibrinogen coagulation. In the 1940s a true golden age in coagulation was started with the results of Quick's prothrombin time test, described in 1936, and since then the most employed coagulation test. Besides numerous factors involved in this function were discovered and classified. The discovery and introduction of heparin and coumarin in the anticoagulant therapeutics opened a measureless panorama for the arrival of new antithrombotic drugs. By the middle of the XX century, the mechanism of coagulation had been practically deciphered and diverse models based on sequential enzymatic function were proposed, called first chain reactions and in cascade reactions later. In the second half of the XX century, numerous regulatory mechanisms of coagulation were identified and diverse laboratory tests appeared that have allowed highly precise diagnoses of a variety of diseases. Blood separation techniques have permitted to produce factor concentrates for clinical use. With the identification of the genes that encode the synthesis of coagulation factors, it has been possible to produce them by means of molecular biology techniques, being the most significant the production of factors VIII, IX, and VII. The present model of coagulation is based on tissular factor activation and the participation of cells, concepts that were already implicit in the classical theory of Morawitz. PMID- 16366179 TI - One on one. PMID- 16366178 TI - [Levosimendan: a new strategy in the treatment of heart failure]. AB - Congestive heart failure is a long standing health issue. Traditionally, heart failure has been treated with a wide array of drugs such as diuretics, digitalis, catecholamine and non catecholamine inotropics, although treatment with these drugs bears adverse effects, such as the generation of arrhythmia and even death. A new class of drugs has recently exerted a positive impact on the treatment of patients with heart failure; these are the calcium sensitizers that enhance myocardial contractility without increasing cytosolic calcium. Levosimendan is a calcium sensitizer that, besides increasing contractility, has a vasodilating effect due to the activation of K(ATP) channels, being both mechanisms responsible for an advantageous therapeutic option. Different studies have proven the efficiency and safety profile of the drug on various scenarios and populations; thereby considering levosimendan a real and safe alternative treatment for patients with acute or chronic ventricular failure that need intravenous pharmacological support. PMID- 16366180 TI - Getting the upper hand. PMID- 16366181 TI - Team solution. PMID- 16366182 TI - Optional but necessary. PMID- 16366183 TI - Gate control. PMID- 16366184 TI - Building bridges. PMID- 16366185 TI - 'Tis the season for legal checks 'n' balances. PMID- 16366186 TI - Creating a doctoring profession. PMID- 16366187 TI - Challenges to confront diabetic retinopathy and retinopathy of prematurity. PMID- 16366188 TI - Amniotic membrane transplantation in ocular surface disorders. AB - In this prospective study, 81 eyes of 70 patients diagnosed with various ocular surface disorders were enrolled to document the use of amniotic membrane transplantation in various ocular surface disorders. Detailed history and ocular examination was done. Ocular photographs and consent from all patients were taken. Fluorescein staining and impression cytology was done preoperatively and postoperatively in selected cases. Amniotic membrane was prepared from the placenta of a donor (consent taken and negative for infectious disorders), after separating amnion from chorion. It was washed with antibiotic solutions, transferred over nitrocellulose paper and stored in Dulbecco's modified Eagle's minimum essential medium at -80 degrees C. Recipient bed was prepared by removing the fibrovascular pannus and necrosed conjunctiva. Amniotic membrane was transplanted with the epithelial side up and sutured. Sixty-four eyes had good result by clinical evaluation or impression cytology findings, 5 eyes later required limbal stem cell culture and transplantation. All the 3 eyes had failure of the fornix reconstruction and 5 eyes had recurrence of the pterygium. Amniotic membrane provides lower recurrence rate in cases of recurrent pterygium. Alkali injuries are more dangerous but showed good response to amniotic membrane transplantation combined with limbal autografting or ex-vivo expansion and later transfer. Initial proper assessment of limbal involvement, conjunctival necrosis and corneal involvement is the key to the management of acute cases. Contracted sockets showed no improvement. Shield ulcers and persistent epithelial defect and ocular surface defects secondary to tumour excision showed excellent results. PMID- 16366189 TI - Retinopathy of prematurity. AB - Retinopathy of prematurity (ROP) is a vasoproliferative disorder of the retina that occurs in premature and low birth weight babies. Stages of ROP are 1, 2, 3, 4A, 4B and 5. There are three zones I, II and III. Screening is essential to detect ROP. Whom, when and how to screen ROP has been narrated in a nutshell. Once threshold ROP is detected, it is essential to treat within 72 hours. Retinal ablation of the avascular retina is done using laser photocoagulation or cryotherapy. Rush disease is primarily a zone I ROP with extraretinal proliferation. It requires aggressive laser treatment with larger number of high intensity closely applied spots to stop the progression. Surgical management is advocated in stages IVA and IVB ROP by scleral buckling. Stage V requires complex vitreoretinal surgery. It is important to promote awareness about the disease and to ensure timely management to prevent blindness. PMID- 16366190 TI - A study on micro-albuminuria--an independent risk factor for vasculopathy in diabetes mellitus. AB - Diabetes is becoming a serious threat to combat felt among the doctors of the world. India, in partcular is a vulnerable country contributing maximum number of cases to the global diabetic pool. Macro-and microvascular complications are important causes of mortality and morbidity. Micro-albuminuria is a surrogate marker for detetion of vasculopathy for which early detection and aggressive treatment can reduce mortality and morbidity. A small study was conducted to identify the usefulness of this marker in our setting which can be used as a cost effective tool for detecting the dreadful complication early. This study has demonstrated that in presence of micro-albuminuria the vascular complications of diabetes definitely increase and it is more relevant in cases of type 2 diabetes irrespective of other parameters rendering it to be a independent risk factor. It also indicates that presence of this marker along with vasculopathy is time dependent ie, more the duration of the disease more is the complication. PMID- 16366191 TI - Ocular involvement in diabetes mellitus. AB - Diabetes is a worldwide medical problem and is a significant cause of morbidity and mortality. It has considerable impact on both the patient and the society because it typically affects individuals in their most productive years. It is also one of the leading causes of blindness and visual impairment. A person with diabetes has 25 times the risk of blindness compared to a non-diabetic. This article reviews the variety of ways in which the eye and its adnexa can be involved in diabetes mellitus. PMID- 16366192 TI - Orbital rhinosporidiosis. AB - Rhinospordiosis, a chronic granulomatous disease of the mucocutaneous tissue, is endemic in India and other parts of South East Asia. It commonly affects the upper respiratory tract, but involvement of other sites has also been reported. A 27-year-old male patient reported with features simulating severe obital cellulitis underwent emergency managemt for his systemic problems. Subsequent CT scan and histopathological examination confirmed the diagnosis of rhinoporidiois. Definitive surgical treatment was done to remove the suspected papillomatous mass which led to marked improvement of the patient. PMID- 16366193 TI - Hurler's disease. AB - Hurler's disease, also known as mucopolysaccharidoses 1H, an autosomal recessive disease due to enzyme alpha-iduronidase deficiency has generally a spectrum of findings involving the eye, skeletal abnormality, organomegaly, joint stiffness, hernia, mental retardation and cardiovascular abnormalities. The two cases presented here are related to each other as brother of 9 years age and sister of 7 years of age with their parents had consanguineous marriage. On ocular examination in both the cases there was dense bilateral corneal opacities. Glycosaminoglycan's level was found to be 4 mg/mmol cr in the first case and 5 mg/mmol cr in the second case. In the first case no ocular treatment was planned and in the second case penetrating keratoplasty was the choice of management. PMID- 16366194 TI - Study of efficacy and safety of losartan (losar) in Asian Indian hypertensives. AB - Although there has been widespread dissemination of knowledge about hypertension, it remains poorly treated in most populations. Systemic hypertension is associated with increased risk for coronary artery disease, stroke, nephrosclerosis, peripheral vascular disease, etc. The treatment of hypertension includes non-pharmacological measures and the specific drug therapy. Losartan potassium is an orally active, non-peptide angiotensin II receptor antagonist. It is the first of this new class of drugs introduced for clinical use in hypertension. Data was obtained of 347 patients from 140 general physicians. The study revealed that losartan potassium is used in the treatment of mild to moderate hypertension with excellent to good response in 98.8% of the cases. Mild adverse reactions were reported in 5.8% of the cases. None of the adverse reactions were severe enough which required discontinuation of therapy or needed hospitalisation. Thus, the present postmarketing surveillance study confirms the safety and efficacy of losartan potassium in Indian population. PMID- 16366195 TI - Comparative randomised active drug controlled clinical trial of a herbal eye drop in computer vision syndrome. AB - A comparative double-blind placebo-controlled clinical trial of a herbal eye drop (itone) was conducted to find out its efficacy and safety in 120 patients with computer vision syndrome. Patients using computers for more than 3 hours continuously per day having symptoms of watering, redness, asthenia, irritation, foreign body sensation and signs of conjunctival hyperaemia, corneal filaments and mucus were studied. One hundred and twenty patients were randomly given either placebo, tears substitute (tears plus) or itone in identical vials with specific code number and were instructed to put one drop four times daily for 6 weeks. Subjective and objective assessments were done at bi-weekly intervals. In computer vision syndrome both subjective and objective improvements were noticed with itone drops. Itone drop was found significantly better than placebo (p<0.01) and almost identical results were observed with tears plus (difference was not statistically significant). Itone is considered to be a useful drug in computer vision syndrome. PMID- 16366196 TI - Anaemia free India campaign. PMID- 16366197 TI - The effect of cold exposure on the hormonal and metabolic responses to sleep deprivation. AB - OBJECTIVE: The purpose of this study was to determine the effect of 12 degrees C cold exposure for 180-minutes on the hormonal responses of sleep-deprived individuals. METHODS: Participants underwent 2 cold-air trials: 1 after a normal night of sleep (ie, 6-8 hours) and 1 after 33 hours of sleep deprivation (SDEP). A venous blood sample was taken at baseline and then at 90-and 180-minute cold exposure time points. Repeated-measures analysis of variance was used to determine significance between a normal night of sleep and SDEP for norepinephrine, epinephrine, cortisol, insulin, thyroid hormones triiodothyronine and thyroxine, glucose, and non-esterifled fatty acids. RESULTS: There was no significant main effect for time, trial, or interaction for insulin, thyroid hormones, epinephrine, cortisol, and glucose (P < or = .05). A significant main effect for time for norepinephrine and non-esterified fatty acids was demonstrated (P < .001). DISCUSSION: The lack of significant differences in the hormonal and metabolic responses to cold exposure combined with SDEP may have been because of an ability of the individual to continue to respond despite the environmental stressor or the physiological effect elicited from cold exposure, thereby possibly masking physiological responses of SDEP. CONCLUSIONS: On the basis of these data, SDEP combined with protracted cold exposure apparently was not a great enough stressor to cause a differential response in the hormonal and metabolic parameters. PMID- 16366198 TI - Unique occupational hazards of Alaska: animal-related injuries. AB - OBJECTIVE: During 1992-2000, an average of 40 fatal occupational injuries and 12,400 nonfatal occupational injuries and illnesses related to animals were recorded each year in the United States, most involving domestic farm animals. Although Alaska has a relatively small farming industry, it supports several industries that require workers to regularly be in contact with animals. This study examines the pattern and characteristics of animal-related occupational injuries in Alaska. METHODS: Two data sources were accessed: the Alaska Trauma Registry for nonfatal injuries requiring hospitalization and the Alaska Occupational Injury Surveillance System for fatal injuries. The case definition included events in which the source of the injury was an animal or animal product (Occupational Injury and Illness Classification Manual source code 51). RESULTS: In Alaska during 1991-2000, there were 43 animal-related occupational injuries requiring hospitalization and 25 animal-related fatalities. There were only 2 fatal events: 1 bird-strike aircraft accident killing 24 military personnel and 1 bear attack. The majority of the nonfatal injury events were related to marine wildlife (n = 20), with the rest related to either domesticated (n = 11) or nondomesticated (n = 12) mammals. Of events reporting a hospital charge (23 of 43), the average cost was over dollar 9700 per person. CONCLUSIONS: The catastrophic aircraft crash increased bird-control efforts near airports around the state. The nonfatal animal-related injuries have received less notice, although they result in thousands of dollars in hospital costs and lost workdays. Fishing-industry workers in particular should be made aware of potential injuries and educated on how to treat them when away from definitive medical care. PMID- 16366199 TI - Recreational injuries in Washington state national parks. AB - OBJECTIVES: The objectives of this study were to identify the number and types of recreational injuries sustained by visitors to Mount Rainier National Park and Olympic National Park in Washington State and to compare the nature of injuries sustained by children compared with adults. METHODS: We retrospectively reviewed case incident reports obtained by rangers in Mount Rainer National Park and Olympic National Park between 1997 and 2001. Data collected included victim age, gender, date of injury, activity preinjury, type of injury, and mechanism of injury. RESULTS: There were 535 cases of recreational wilderness injuries (including 19 total deaths), yielding a rate of 22.4 injuries per million visits. The mean age of injury victims was 34 years. Males were more likely to sustain injury than were females (59% vs 41%). Most injuries occurred during summer months between noon and 6:00 PM, and 90% occurred during daylight hours. The most common preinjury activities included hiking (55%), winter sports (15%), and mountaineering (12%), and the most common types of injuries included sprains, strains and soft tissue injuries (28%), fractures or dislocations (26%), and lacerations (15%). A total of 121 (23%) of the injuries occurred in children (<18 years of age). There were 19 deaths in the 2 national parks (18 men, 1 woman); all victims were adults. Hiking (58%) and mountaineering (26%) were the most common activities at the time of death. Mechanism of death included falls (37%), medical (eg, myocardial infarction) (21%), drowning (5%), and suicide (5%). CONCLUSIONS: The most common type of injury was soft tissue injury, and injuries occurred most commonly while hiking, during daylight hours, and in the summer. Preinjury activities and types of injuries were different in children compared with adults. Knowledge of how and when injuries occur in national parks can assist in determining what resources are needed to help provide a safer environment for park visitors. This study may also aid prevention strategies in the national parks, guide training of rangers, aid in the preparation of first aid kits, and further the education of people who participate in wilderness activities. PMID- 16366200 TI - Physiological responses of ultraendurance athletes and nonathletes during an attempt to summit Denali. AB - OBJECTIVE: To compare altitude responses of 2 ultraendurance athletes and 2 nonathletes during a 2-week expedition on Denali (Mount McKinley). METHODS: The severity of acute mountain sickness (AMS) symptoms (Lake Louise AMS guidelines) and pulmonary function parameters (forced vital capacity, forced expiratory volume in 1 second, peak expiratory flow) as well as resting heart rate and arterial oxygen saturation measurements were taken during the climb. Baseline measurements were made at 375 m, and field tests were performed at altitudes of 2200 m, 2400 m, 3000 m, 3400 m, 4100 m, 4300 m, and 10 m. RESULTS: Nonathletes reported moderate AMS symptoms at altitudes up to and including 3000 m, whereas ultraendurance athletes reported moderate AMS symptoms at altitudes above 3000 m. Considerable daily variation existed in pulmonary function measures within and between groups; however, the largest shift from baseline and between groups occurred at 3000 m where ultraendurance athletes had increased and nonathletes had decreased peak expiratory flow and forced vital capacity. Resting heart rate increased and arterial oxygen saturation decreased with altitude. CONCLUSIONS: Highly aerobically fit individuals may be more susceptible to delayed and more prolonged onset of AMS than are moderately fit individuals. Pulmonary function, although highly variable, also may be dissimilar between these groups. PMID- 16366201 TI - Puncture wounds by driftwood catfish during bucket baths: local habits of riverside people and fish natural history in the Amazon. AB - OBJECTIVE: To assess the prevalence of stings by small spiny driftwood catfish (caratai) of the genus Centromochlus (Auchenipteridae) accidentally caught in buckets during bucket bathing by riverside people along the Brazilian Amazon and to determine the probability of catching specimens of these fish during random throws of a bucket into the river. METHODS: We interviewed 27 adult residents living at the confluence of the Negro and Solimoes rivers in Brazil regarding whether or not they had ever been stung by driftwood catfish while bucket bathing. To assess the likelihood of catching catfish in bathing buckets, we randomly threw a typical plastic bucket used for bathing in 4 series of 10 throws into the river at dusk or night around a floating house. RESULTS: Seventeen of the 27 subjects (63%) reported being injured by driftwood catfish during bucket bathing. Three individuals (17.6%) had been injured 2 to 3 times, totaling 23 puncture accidents. All stings occurred at dusk or early night. In the 4 series of 10 bucket throws, we caught 3 driftwood catfish (in 1 series we did not catch any fish). Thus, the chance of catching a driftwood catfish in a single bucket throw at dusk was slightly less than 10%. CONCLUSIONS: The prevalence of stings by driftwood catfish to people bucket bathing in this section of the Brazilian Amazon is high, partly because of the relatively high chances of catching these small catfish during random throws of a bathing bucket into the river. PMID- 16366202 TI - Resuscitation from hemorrhagic shock using rectally administered fluids in a wilderness environment. AB - We report the successful use in a wilderness environment of rectally administered oral rehydation fluid to resuscitate a patient who was in shock. The subject was a 21-year-old Nepali man who had experienced a major upper gastrointestinal hemorrhage. PMID- 16366203 TI - Difficult decisions at altitude: the management of an acutely dyspneic porter at 5000 meters. AB - We report a case of a porter who presented with acute dyspnea during an expedition in Nepal at 5000 m above sea level. We present the difficulties involved in making the correct diagnosis in this patient under austere conditions and discuss the difficult decisions that must be made in caring for such a patient. PMID- 16366204 TI - Water intoxication: a possible complication during endurance exercise--a commentary. PMID- 16366206 TI - Fat desire and fat intake at extreme altitude: observations from the "On Top Everest '89" Nutrition Research Expedition. PMID- 16366205 TI - Water intoxication: a possible complication during endurance exercise, 1985. PMID- 16366207 TI - Pacemakers and avalanche transceivers: a cautionary tale. PMID- 16366208 TI - Will a 'perfect storm' help control rising orthopedic implant costs? PMID- 16366209 TI - More ORs call for financial disclosure with product, equipment requests. PMID- 16366210 TI - VHA acquires firm known for gainsharing. PMID- 16366211 TI - Katrina teaches disaster planning lessons. PMID- 16366212 TI - Mix shifts toward high-demand implants. PMID- 16366213 TI - Matrix caps pricing for ortho implants. PMID- 16366214 TI - A small hospital tackles implant costs. PMID- 16366215 TI - A strategy for review of new products. PMID- 16366216 TI - Should ASCcs hire RNs from the hospital? PMID- 16366217 TI - A plan for managing problem employees. PMID- 16366218 TI - The development of a Master of Public Health Program with an initial focus on urban and immigrant health at the State University of New York, Downstate Medical Center. AB - The State University Downstate Medical Center initiated a Master of Public Health (MPH) degree program in July 2001 following planning efforts that began in 1995. Twelve Students entered the program in June 2002. Currently, eighty students are enrolled in the program and eighteen have graduated from it in 2004 and 2005. With an initial focus on urban and immigrant health, the program aims to train public health professionals who can assist in addressing through population-based interventions the health issues of Brooklyn's 2,465,326 people, of whom 38.5% are immigrants to the United States. Starting with four courses in the summer 2002 semester, the program now offers twenty-four courses over the three semesters of the academic year. The program is housed in the Department of Preventive Medicine and Community Health of the College of Medicine and is part-time in nature for most students. In addition to completing required course work, students must also complete a 250-hour practicum experience in which they apply theoretical knowledge in a public health practice setting. Student practicum experiences play a vital role in linking the program to communities and serve as conduits for the initiation of further community based collaboratives. This article describes the challenges encountered in initiating an MPH program in an academic medical center, the importance of both intramural and community support to its success, and the vital role it plays in addressing the health issues of various communities. The program became a leading priority of the Strategic Plan of the Downstate Medical Center in 2000, and received the full support of Downstate's then new president, Dr. John C. LaRosa. This prioritization and support proved essential to the rapid development of the program. The Downstate MPH program offers a concurrent degree to medical students who are able to complete both degrees in a four year period. The Alumni Fund of the College of Medicine provides each MD/MPH student with a one-time scholarship which covers a quarter of the MPH tuition. Concurrent MPH degrees are also offered for graduate students enrolled in occupational therapy, nursing, and several other health programs. The Council on Education for Public Health (CEPH) conducted an accreditation site visit of the Downstate MPH program in December 2004. On June 10, 2005, the CEPH Board accredited the program for 5 years. PMID- 16366219 TI - Travel-related savings through a rural, clinic-based automated drug dispensing system. AB - For rural patients, access to pharmacies is often impeded by geographical and financial factors. The number of available pharmacies may be limited, and the extensive travel needed to access them is time consuming, potentially dangerous and, due to rising fuel prices, increasingly expensive. We describe the benefits of a clinic-based, automated medication dispensing system (ADDS) implemented in two rural communities in the Adirondack Mountain region of New York State. To estimate distance saved by utilizing the ADDS at the clinic, data from 268 consecutive patient encounters was imported into a mapping software package. Clinic Trip and Pharmacy Trip distances were calculated, with Miles saved estimated as the difference. Cost savings estimates were generated, with results reported as percentage of median per capita money income by county. Patients lived significantly closer to both clinics than to nearest traditional pharmacies, and saved an average travel distance of 32.6 and 18.0 miles per encounter. Cost estimates suggest travel-related savings of dollar 12.23 and dollar 6.75 per encounter, or 3.4% and 1.7% of weekly income. Considering the median out of pocket expense (dollar 13.31) for acquiring a prescription, patients traveling more than 35 miles round trip to obtain medications spent more on travel than on the medication itself. Placement of ADDS can significantly reduce the travel distances required for drug procurement in rural communities, and can provide acutely needed medications without travel-related delay. Such systems can provide considerable travel-related savings to the rural poor, where travel-related costs can exceed average drug costs. PMID- 16366220 TI - Sun protection policies and practices at child care centers in Massachusetts. AB - We assessed the relationship between sun protection policies and practices at child care centers in Massachusetts. We hypothesized that centers with sun protection policies were more likely to have regular sun protection practices in place compared to centers without these policies. We conducted a telephone survey with directors or assistant directors at 327 child care centers during the summer of 2002. The main outcome measure was sun protection practices, which included time spent outside during mid-day and the use of sunscreen, hats, and protective clothing by the majority of children assessed over the last 5 program days. The 36-item survey also inquired about the center's sun protection policy and included demographic questions. Most centers (73%) reported having a written sun protection policy. Sun protection policies were positively associated with reported sunscreen (chi squared = 14.63, p = 0.0001) and hat use (chi squared = 30.98, p < 0.0001) and inversely associated with time outside (chi squared = 10.76, p = 0.001). Seventy-seven percent of centers followed recommended sunscreen practices. However, centers were far less likely to have recommended hat use (36%) and protective clothing (1.5%) practices. A formal sun protection policy may be an effective way to increase sun protection practices in the child care setting. Further research should assess this relationship in other states. Improving and expanding existing state regulations may be a reasonable strategy to increase sun protection at child care centers. PMID- 16366221 TI - Modeling robust oscillatory behavior of the hypothalamic-pituitary-adrenal axis. AB - A mathematical model of the hypothalamic-pituitary-adrenal (HPA) axis of the human endocrine system is proposed. This new model provides an improvement over previous models by introducing two nonlinear factors with physiological relevance: 1) a limit to gland size; 2) rejection of negative hormone concentrations. The result is that the new model is by far the most robust; e.g., it can tolerate at least -50% and +100% perturbations to any of its parameters. This high degree of robustness allows one, for the first time, to model features of the system such as circadian rhythm and response to hormone injections. In addition, relative to its closest predecessor, the model is simpler; it contains only about half of the parameters, and yet achieves more functions. The new model provides opportunities for teaching endocrinology within a biological or medical school context; it may also have applications in modeling and studying HPA axis disorders, for example, related to gland size dynamics, abnormal hormone levels, or stress influences. PMID- 16366222 TI - A model for surface EMG generation in volume conductors with spherical inhomogeneities. AB - Most models for surface electromyography (EMG) signal generation are based on the assumption of space-invariance of the system in the direction of source propagation. This assumption implies the same shape of the potential distribution generated by a source in any location along the propagation direction. In practice, the surface EMG generation system is not space invariant and, therefore, the surface signal detected along the direction of the muscle fibers may significantly change shape along the propagation path. An important class of nonspace invariant systems is that of volume conductors inhomogeneous in the direction of source propagation. In this paper, we focused on inhomogeneities introduced by the presence of spheres of different conductivities with respect to the tissue where they are located. This effect may prove helpful to model the presence of glands, vessels, or local changes in the conductivity of a tissue. We present an approximate analytical solution that accounts for an arbitrary number of spheres in an arbitrary complex volume conductor. As a representative example, we propose the solution for a planar layered volume conductor, comprised of fat and muscle layers with spherical inhomogeneities inside the fat layer. The limitations of the approximations introduced are discussed. The model is computationally fast and constitutes an advanced means for the analysis and interpretation of surface EMG signal features. PMID- 16366223 TI - Comprehensive pharmacokinetic model of insulin Glargine and other insulin formulations. AB - In this paper, a comprehensive pharmacokinetic model for different insulin formulations including insulin Glargine is developed based on the model proposed by Trajanoski et al. (1993). Current models show limitations for insulin Glargine due to the appearance of an uncharacteristic peak in the concentration-time evolution of plasma insulin that does not coincide with real experimental data. This important limitation has been solved in this paper by introducing a new virtual insulin state called the bound state, in addition to the dimeric and hexameric ones. Trying to describe the retarded action of insulin Glargine, the modeling idea behind this approach is that immediately after the subcutaneous injection all the insulin resides in the bound state, and only then small amounts of insulin in the hexameric form disengage from the bound state. For the model evaluation different simulation results are compared. Using experimental data published by Lepore et al. (2000), the developed model turned out to be capable of at least qualitatively predicting the concentration-time profile of plasma insulin. Both exogenous insulin flow simulations and spatial diffusion simulations show the plausibility and correct implementation of the derived model. Considering all these simulation results, the here presented new pharmacokinetic model demonstrates to be able to reproduce real patient behavior simulating even complete insulin regimes including long-acting, intermediate and short-acting insulin formulations. PMID- 16366224 TI - Modeling common dynamics in multichannel signals with applications to artifact and background removal in EEG recordings. AB - Removing artifacts and background electroencephaloraphy (EEG) from multichannel interictal and ictal EEG has become a major research topic in EEG signal processing in recent years. We applied for this purpose a recently developed subspace-based method for modeling the common dynamics in multichannel signals. When the epileptiform activity is common in the majority of channels and the artifacts appear only in a few channels the proposed method can be used to remove the latter. The performance of the method was tested on simulated data for different noise levels. For high noise levels the method was still able to identify the common dynamics. In addition, the method was applied to real life EEG recordings containing interictal and ictal activity contaminated with muscle artifact. The muscle artifacts were removed successfully. For both the synthetic data and the analyzed real life data the results were compared with the results obtained with principal component analysis (PCA). In both cases, the proposed method performed better than PCA. PMID- 16366225 TI - Real-time separation of perfusion and oxygenation signals for an implantable sensor using adaptive filtering. AB - In this paper, an adaptive filtering algorithm to separate signals due to perfusion and oxygenation has been developed using an 810-nm source, in addition to 660-nm and 940-nm sources, as an internal reference due to its limited oxygen sensitivity. The newly developed algorithm was tested using Monte Carlo simulated data to prove the effectiveness of the 810-nm reference and adaptive algorithm. Following the simulation, an in vitro model was developed to test the algorithm that used a blood flow through system wrapped with tissue. The system had the ability to isolate the effects of perfusion and oxygenation and the algorithm accurately captured the changes in these signals with reliable consistency. Using the serosal surface of the swine jejunum, in vivo data was also taken to analyze the algorithms response to fluctuating perfusion levels like that seen in hemorrhaging or failing transplants. The algorithm was able to extract the perfusion information from the oxygenation information in this in vivo study. Overall, it was shown that an adaptive filtering algorithm using an 810-nm reference has provided a means to separate oxygenation and perfusion. PMID- 16366226 TI - Current density imaging and electrically induced skin burns under surface electrodes. AB - The origin of electrical burns under gel-type surface electrodes is a controversial topic that is not well understood. To investigate the phenomenon, we have developed an excised porcine skin-gel model, and used low-frequency current density imaging (LFCDI) to determine the current density (CD) distribution through the skin before and after burns were induced by application of electrical current (200 Hz, 70% duty cycle, 20-35 mA monophasic square waveform applied to the electrodes for 30-135 min). The regions of increased CD correlate well with the gross morphological changes (burns) observed. The measurement is sensitive enough to show regions of high current densities in the pre-burn skin, that correlate with areas were burn welts were produced, thus predicting areas where burns are likely to occur. Statistics performed on 28 skin patches revealed a charge dependency of the burn areas and a relatively uniform distribution. The results do not support a thermal origin of the burns but rather electro-chemical mechanisms. We found a statistically significant difference between burn area coverage during anodic and cathodic experiments. PMID- 16366227 TI - Iris-based cyclotorsional image alignment method for wavefront registration. AB - In refractive surgery, especially wavefront-guided refractive surgery, correct registration of the treatment to the cornea is of paramount importance. The specificity of the custom ablation formula requires that the ablation be applied to the cornea only when it has been precisely aligned with the mapped area. If, however, the eye has rotated between measurement and ablation, and this cyclotorsion is not compensated for, the rotational misalignment could impair the effectiveness of the refractive surgery. To achieve precise registration, a noninvasive method for torsional rotational alignment of the captured wavefront image to the patient's eyes at surgery has been developed. This method applies a common coordinate system to the wavefront and the eye. Video cameras on the laser and wavefront devices precisely establish the spatial relationship between the optics of the eye and the natural features of the iris, enabling the surgeon to identify and compensate for cyclotorsional eye motion, whatever its cause. PMID- 16366228 TI - A hierarchical statistical modeling approach for the unsupervised 3-D biplanar reconstruction of the scoliotic spine. AB - This paper presents a new and accurate three-dimensional (3-D) reconstruction technique for the scoliotic spine from a pair of planar and conventional (postero anterior with normal incidence and lateral) calibrated radiographic images. The proposed model uses a priori hierarchical global knowledge, both on the geometric structure of the whole spine and of each vertebra. More precisely, it relies on the specification of two 3-D statistical templates. The first, a rough geometric template on which rigid admissible deformations are defined, is used to ensure a crude registration of the whole spine. An accurate 3-D reconstruction is then performed for each vertebra by a second template on which nonlinear admissible global, as well as local deformations, are defined. Global deformations are modeled using a statistical modal analysis of the pathological deformations observed on a representative scoliotic vertebra population. Local deformations are represented by a first-order Markov process. This unsupervised coarse-to-fine 3-D reconstruction procedure leads to two separate minimization procedures efficiently solved in our application with evolutionary stochastic optimization algorithms. In this context, we compare the results obtained with a classical genetic algorithm (GA) and a recent Exploration Selection (ES) technique. This latter optimization method with the proposed 3-D reconstruction model, is tested on several pairs of biplanar radiographic images with scoliotic deformities. The experiments reported in this paper demonstrate that the discussed method is comparable in terms of accuracy with the classical computed-tomography-scan technique while being unsupervised and while requiring only two radiographic images and a lower amount of radiation for the patient. PMID- 16366229 TI - 3-D reconstruction of microcalcification clusters using stereo imaging: algorithm and mammographic unit calibration. AB - The three-dimensional (3-D) shape of microcalcification clusters is an important indicator in early breast cancer detection. In fact, there is a relationship between the cluster topology and the type of lesion (malignant or benign). This paper presents a 3-D reconstruction method for such clusters using two 2-D views acquired during standard mammographic examinations. For this purpose, the mammographic unit was modeled using a camera with virtual optics. This model was used to calibrate the acquisition unit and then to reconstruct the clusters in the 3-D space after microcalcification segmentation and matching. The proposed model is hardware independent since it is suitable for digital mammographic units with different geometries and with various physical acquisition principles. Three dimensional reconstruction results are presented here to prove the validity of the method. Tests were first performed using a phantom with a well-known geometry. The latter contained X-ray opaque glass balls representing microcalcifications. The positions of these balls were reconstructed with a 16.25 microm mean accuracy. This very high inherent algorithm accuracy is more than enough for a precise 3-D cluster representation. Further validation tests were carried out using a second phantom including a spherical cluster. This phantom was built with materials simulating the behavior of both mammary tissue and microcalcifications toward Xrays. The reconstructed shape was effectively spherical. Finally, reconstructions were carried out for real clusters and their results are also presented. PMID- 16366230 TI - Automatic intrinsic DNA curvature computation from AFM images. AB - Critical information on several biological processes such as DNA-protein interactions and DNA transcription can be derived from analysis of DNA curvature. Under thermal perturbation, the curvature is composed of static and dynamic contributions, thus, can be described as the sum of intrinsic curvature and a fluctuation contribution. Without considering thermal agitations, the DNA curvature is reducible to the intrinsic component, which is a function of the DNA nucleotide sequence only. In this paper, we present an automated algorithm to determine the DNA intrinsic curvature profiles and the molecular spatial orientations in Atomic Force Microscope images. The algorithm allows to reconstruct the intrinsic curvature profile by filtering the thermal contribution. It detects fragment orientation on atomic force microscope images without labels with a percentage of correct molecular-orientation detection of 96.79% in computer-generated benchmarks, for molecules with a high curvature peak. The automated algorithm reconstructs the intrinsic curvature profile of DNA molecules with a mean square error of 3.8122 x 10(-4) rads over a profile with a central peak value of 0.196 rads, and 6.1 x 10(-3) rads over a curvature profile with two symmetric peaks of about 0.08 rads. Moreover, it correctly detects the location of the peaks in the molecules with a deviation of about 1% of molecule length. PMID- 16366231 TI - Multifrequency transducer for microemboli classification and sizing. AB - The classification of circulating microemboli as gaseous or particulate matter is essential to establish the relevance of the detected embolic signals. Until now, Doppler techniques have failed to determine unambiguously the nature of circulating microemboli. Recently, a new approach based on the analysis of radio frequency (RF) signal and using the nonlinear characteristics of gaseous bubbles to classify emboli was investigated. The main limitation of these studies was the requirement of two separate transducers for transmission and reception. This paper presents a multi-frequency transducer with two independent transmitting elements and a separate receiving part with a wide frequency band. The transmitting elements are positioned in a concentric design and cover a frequency band between 100 and 600 kHz. The receiving part consists of a polyvinylidene fluoride layer. The new transducer has been tested in vitro using gaseous emboli. It could correctly classify and size air emboli with diameters ranging from 10 microm to 105 microm. PMID- 16366232 TI - Electrode polarization impedance in weak NaCl aqueous solutions. AB - In this paper, we characterize the polarization impedance behavior of several common metals in diluted NaCl solution operated at low current densities. The objective was to provide a useful reference for those wishing to calculate the electrode polarization impedance in diluted NaCl solutions. Serial equivalent resistance (R) and capacitance (C) for silver, aluminum, gold, platinum, and medical stainless-steel were measured as a function of frequency (10(-2)-10(3) Hz) and NaCl concentration (2.4-77.0 mmol/L). The ratio of electrode polarization impedance with respect to the bulk resistance was calculated and plotted against concentration for each metal. Such a ratio shows the effect of the electrode polarization contribution as a function of electrolyte concentration when the bulk resistance of the solution changes. All metals showed a decrease of serial resistance Rp and capacitance Cp as a function of frequency. The medical stainless-steel electrode showed largest impedance values at lower frequencies compared to the other electrodes, and was concentration independent at all frequencies. Aluminum had smallest polarization impedance at low frequencies. Pure gold and platinum behaved similar with the exception that the serial resistance for gold showed a lower value at higher frequencies. PMID- 16366233 TI - Noninvasive measurement of heartbeat, respiration, snoring and body movements of a subject in bed via a pneumatic method. AB - We have developed a noninvasive pneumatics-based system by which to measure heartbeat, respiration, snoring, and body movements of a subject in bed. A thin, air-sealed cushion is placed under the bed mattress of the subject and the small movements attributable to human automatic vital functions are measured as changes in pressure using a pressure sensor having an almost flat frequency response from 0.1 to 5 kHz and a sensitivity of 56 mV/Pa. Using the newly developed system, heartbeat, respiration, apnea, snoring and body movements are clearly measured. In addition, the optimal signal-to-noise (S/N) ratio by which to evaluate the reliability of the heart rate measurement is presented. Heart rates were measured for four different body postures, 13 different subjects, four different bed mattresses, and three different sensor positions. For these measurements, the S/N ratios ranged from 15.9 to 23.5 dB, and so were determined to be reliable. PMID- 16366234 TI - Audio-biofeedback for balance improvement: an accelerometry-based system. AB - This paper introduces a prototype audio-biofeedback system for balance improvement through the sonification using trunk kinematic information. In tests of this system, normal healthy subjects performed several trials in which they stood quietly in three sensory conditions while wearing an accelerometric sensory unit and headphones. The audio-biofeedback system converted in real-time the two dimensional horizontal trunk accelerations into a stereo sound by modulating its frequency, level, and left/right balance. Preliminary results showed that subjects improved balance using this audio-biofeedback system and that this improvement was greater the more that balance was challenged by absent or unreliable sensory cues. In addition, high correlations were found between the center of pressure displacement and trunk acceleration, suggesting accelerometers may be useful for quantifying standing balance. PMID- 16366235 TI - Short- and long-term joint symbolic dynamics of heart rate and blood pressure in dilated cardiomyopathy. AB - Autonomic cardiovascular control involves complex interactions of heart rate and blood pressure. In patients with dilated cardiomyopathy (DCM), this control is impaired and parameters for its quantification might be of prognostic importance. In this paper, we introduce methods based on joint symbolic dynamics (JSD) for the enhanced analysis of heart rate and blood pressure interactions. To assess the coarse-grained dynamics beat-to-beat changes of heart rate and blood pressure are encoded in symbol strings. Subsequently, the distribution properties of short symbol sequences (words) as well as the scaling properties of the whole symbol string are assessed. The comparison of joint symbolic heart rate and blood pressure dynamics in DCM (n = 75) with those in healthy controls (n = 75) showed significant changes. Both, the distribution of words and the scaling properties indicate a loss in heart rate dynamics associated with blood pressure regulation in DCM. In conclusion, the analyses of short- and long-term JSDs provide insights into complex physiological heart rate and blood pressure interactions and furthermore reveal patho-physiological cardiovascular control in DCM. PMID- 16366236 TI - Decomposition of three-dimensional medical images into visual patterns. AB - In this paper, we present a method for the decomposition of a volumetric image into its most relevant visual patterns, which we define as features associated to local energy maxima of the image. The method involves the clustering of a set of predefined bandpass energy filters according to their ability to segregate the different features in the image, thus generating a set of composite-feature detectors tuned to the specific visual patterns present in the data. Clustering is based on a measure of statistical dependence between pairs of frequency features. We will illustrate the applicability of the method to the initialization of a three-dimensional geodesic active model. PMID- 16366237 TI - Supervised nonlinear dimensionality reduction for visualization and classification. AB - When performing visualization and classification, people often confront the problem of dimensionality reduction. Isomap is one of the most promising nonlinear dimensionality reduction techniques. However, when Isomap is applied to real-world data, it shows some limitations, such as being sensitive to noise. In this paper, an improved version of Isomap, namely S-Isomap, is proposed. S-Isomap utilizes class information to guide the procedure of nonlinear dimensionality reduction. Such a kind of procedure is called supervised nonlinear dimensionality reduction. In S-Isomap, the neighborhood graph of the input data is constructed according to a certain kind of dissimilarity between data points, which is specially designed to integrate the class information. The dissimilarity has several good properties which help to discover the true neighborhood of the data and, thus, makes S-Isomap a robust technique for both visualization and classification, especially for real-world problems. In the visualization experiments, S-Isomap is compared with Isomap, LLE, and WeightedIso. The results show that S-Isomap performs the best. In the classification experiments, S-Isomap is used as a preprocess of classification and compared with Isomap, WeightedIso, as well as some other well-established classification methods, including the K nearest neighbor classifier, BP neural network, J4.8 decision tree, and SVM. The results reveal that S-Isomap excels compared to Isomap and WeightedIso in classification, and it is highly competitive with those well-known classification methods. PMID- 16366238 TI - Intelligent robust control for uncertain nonlinear time-varying systems and its application to robotic systems. AB - This paper addresses the problem of designing adaptive fuzzy-based (or neural network-based) robust controls for a large class of uncertain nonlinear time varying systems. This class of systems can be perturbed by plant uncertainties, unmodeled perturbations, and external disturbances. Nonlinear H(infinity) control technique incorporated with adaptive control technique and VSC technique is employed to construct the intelligent robust stabilization controller such that an H(infinity) control is achieved. The problem of the robust tracking control design for uncertain robotic systems is employed to demonstrate the effectiveness of the developed robust stabilization control scheme. Therefore, an intelligent robust tracking controller for uncertain robotic systems in the presence of high degree uncertainties can easily be implemented. Its solution requires only to solve a linear algebraic matrix inequality and a satisfactorily transient and asymptotical tracking performance is guaranteed. A simulation example is made to confirm the performance of the developed control algorithms. PMID- 16366239 TI - A biologically inspired active compliant joint using local positive velocity feedback (LPVF). AB - Starting from studies which revealed that positive feedback is found in the control system for walking in arthropods, we have constructed a new positive feedback driven joint that can be used for solving compliant motion tasks. We propose two different joint constructions each of which shows passive compliance. Based on these joints we introduce three different local positive velocity feedback (LPVF) controllers and discuss their properties in the context of motion generation in closed kinematic chains. The third circuit named undelayed dLPVF is used for the control of a compliant planar manipulator which turns a crank. Our concept is of highly decentralized nature and follows the idea of embodiment. In our case this means that a process which is controlled by LPVF controllers reveals its nature when the controllers interact with this process. PMID- 16366240 TI - Modeling of driver's collision avoidance maneuver based on controller switching model. AB - This paper presents a modeling strategy of human driving behavior based on the controller switching model focusing on the driver's collision avoidance maneuver. The driving data are collected by using the three-dimensional (3-D) driving simulator based on the CAVE Automatic Virtual Environment (CAVE), which provides stereoscopic immersive virtual environment. In our modeling, the control scenario of the human driver, that is, the mapping from the driver's sensory information to the operation of the driver such as acceleration, braking, and steering, is expressed by Piecewise Polynomial (PWP) model. Since the PWP model includes both continuous behaviors given by polynomials and discrete logical conditions, it can be regarded as a class of Hybrid Dynamical System (HDS). The identification problem for the PWP model is formulated as the Mixed Integer Linear Programming (MILP) by transforming the switching conditions into binary variables. From the obtained results, it is found that the driver appropriately switches the "control law" according to the sensory information. In addition, the driving characteristics of the beginner driver and the expert driver are compared and discussed. These results enable us to capture not only the physical meaning of the driving skill but the decision-making aspect (switching conditions) in the driver's collision avoidance maneuver as well. PMID- 16366241 TI - Solving minimum distance problems with convex or concave bodies using combinatorial global optimization algorithms. AB - Determining the minimum distance between convex objects is a problem that has been solved using many different approaches. On the other hand, computing the minimum distance between combinations of convex and concave objects is known to be a more complicated problem. Most methods propose to partition the concave object into convex subobjects and then solve the convex problem between all possible subobject combinations. This can add a large computational expense to the solution of the minimum distance problem. In this paper, an optimization based approach is used to solve the concave problem without the need for partitioning concave objects into convex pieces. Since the optimization problem is no longer unimodal (i.e., has more than one local minimum point), global optimization techniques are used. Simulated Annealing (SA) and Genetic Algorithms (GAs) are used to solve the concave minimum distance problem. In order to reduce the computational expense, it is proposed to replace the objects' geometry by a set of points on the surface of each body. This reduces the problem to an unconstrained combinatorial optimization problem, where the combination of points (one on the surface of each body) that minimizes the distance will be the solution. Additionally, if the surface points are set as the nodes of a surface mesh, it is possible to accelerate the convergence of the global optimization algorithm by using a hill-climbing local optimization algorithm. Some examples using these novel approaches are presented. PMID- 16366242 TI - A Weighted Sum Validity Function for clustering with a Hybrid Niching Genetic Algorithm. AB - Clustering is inherently a difficult problem, both with respect to the construction of adequate objective functions as well as to the optimization of the objective functions. In this paper, we suggest an objective function called the Weighted Sum Validity Function (WSVF), which is a weighted sum of the several normalized cluster validity functions. Further, we propose a Hybrid Niching Genetic Algorithm (HNGA), which can be used for the optimization of the WSVF to automatically evolve the proper number of clusters as well as appropriate partitioning of the data set. Within the HNGA, a niching method is developed to preserve both the diversity of the population with respect to the number of clusters encoded in the individuals and the diversity of the subpopulation with the same number of clusters during the search. In addition, we hybridize the niching method with the k-means algorithm. In the experiments, we show the effectiveness of both the HNGA and the WSVF. In comparison with other related genetic clustering algorithms, the HNGA can consistently and efficiently converge to the best known optimum corresponding to the given data in concurrence with the convergence result. The WSVF is found generally able to improve the confidence of clustering solutions and achieve more accurate and robust results. PMID- 16366243 TI - Texture image retrieval using new rotated complex wavelet filters. AB - A new set of two-dimensional (2-D) rotated complex wavelet filters (RCWFs) are designed with complex wavelet filter coefficients, which gives texture information strongly oriented in six different directions (45 degrees apart from complex wavelet transform). The 2-D RCWFs are nonseparable and oriented, which improves characterization of oriented textures. Most texture image retrieval systems are still incapable of providing retrieval result with high retrieval accuracy and less computational complexity. To address this problem, we propose a novel approach for texture image retrieval by using a set of dual-tree rotated complex wavelet filter (DT-RCWF) and dual-tree-complex wavelet transform (DT-CWT) jointly, which obtains texture features in 12 different directions. The information provided by DT-RCWF complements the information generated by DT-CWT. Features are obtained by computing the energy and standard deviation on each subband of the decomposed image. To check the retrieval performance, texture database D1 of 1856 textures from Brodatz album and database D2 of 640 texture images from VisTex image database is created. Experimental results indicates that the proposed method improves retrieval rate from 69.61% to 77.75% on database D1, and from 64.83% to 82.81% on database D2, in comparing with traditional discrete wavelet transform based approach. The proposed method also retains comparable levels of computational complexity. PMID- 16366244 TI - Dynamic algorithms for the shortest path routing problem: learning automata-based solutions. AB - This paper presents the first Learning Automaton-based solution to the dynamic single source shortest path problem. It involves finding the shortest path in a single-source stochastic graph topology where there are continuous probabilistic updates in the edge-weights. The algorithm is significantly more efficient than the existing solutions, and can be used to find the "statistical" shortest path tree in the "average" graph topology. It converges to this solution irrespective of whether there are new changes in edge-weights taking place or not. In such random settings, the proposed learning automata solution converges to the set of shortest paths. On the other hand, the existing algorithms will fail to exhibit such a behavior, and would recalculate the affected shortest paths after each weight-change. The important contribution of the proposed algorithm is that all the edges in a stochastic graph are not probed, and even if they are, they are not all probed equally often. Indeed, the algorithm attempts to almost always probe only those edges that will be included in the shortest path graph, while probing the other edges minimally. This increases the performance of the proposed algorithm. All the algorithms were tested in environments where edge-weights change stochastically, and where the graph topologies undergo multiple simultaneous edge-weight updates. Its superiority in terms of the average number of processed nodes, scanned edges and the time per update operation, when compared with the existing algorithms, was experimentally established. The algorithm can be applicable in domains ranging from ground transportation to aerospace, from civilian applications to military, from spatial database applications to telecommunications networking. PMID- 16366245 TI - Modeling and deadlock avoidance of automated manufacturing systems with multiple automated guided vehicles. AB - An automated manufacturing system (AMS) contains a number of versatile machines (or workstations), buffers, an automated material handling system (MHS), and is computer-controlled. An effective and flexible alternative for implementing MHS is to use automated guided vehicle (AGV) system. The deadlock issue in AMS is very important in its operation and has extensively been studied. The deadlock problems were separately treated for parts in production and transportation and many techniques were developed for each problem. However, such treatment does not take the advantage of the flexibility offered by multiple AGVs. In general, it is intractable to obtain maximally permissive control policy for either problem. Instead, this paper investigates these two problems in an integrated way. First we model an AGV system and part processing processes by resource-oriented Petri nets, respectively. Then the two models are integrated by using macro transitions. Based on the combined model, a novel control policy for deadlock avoidance is proposed. It is shown to be maximally permissive with computational complexity of O (n2) where n is the number of machines in AMS if the complexity for controlling the part transportation by AGVs is not considered. Thus, the complexity of deadlock avoidance for the whole system is bounded by the complexity in controlling the AGV system. An illustrative example shows its application and power. PMID- 16366246 TI - Reachability of nonsynchronized choice Petri nets and its applications. AB - A new local structure called a second-order structure was proposed to generate a new class of nets called synchronized choice nets (SNC). SNC covers well-behaved free choice nets. Reachability is no longer P-Space hard problem, but can be solved with polynomial time complexity. How to extend them to non-SNC and its application to deadlock detection in flexible manufacturing systems are discussed. PMID- 16366247 TI - A layered goal-oriented fuzzy motion planning strategy for mobile robot navigation. AB - Most conventional motion planning algorithms that are based on the model of the environment cannot perform well when dealing with the navigation problem for real world mobile robots where the environment is unknown and can change dynamically. In this paper, a layered goal-oriented motion planning strategy using fuzzy logic is developed for a mobile robot navigating in an unknown environment. The information about the global goal and the long-range sensory data are used by the first layer of the planner to produce an intermediate goal, referred to as the way-point, that gives a favorable direction in terms of seeking the goal within the detected area. The second layer of the planner takes this way-point as a subgoal and, using short-range sensory data, guides the robot to reach the subgoal while avoiding collisions. The resulting path, connecting an initial point to a goal position, is similar to the path produced by the visibility graph motion planning method, but in this approach there is no assumption about the environment. Due to its simplicity and capability for real-time implementation, fuzzy logic has been used for the proposed motion planning strategy. The resulting navigation system is implemented on a real mobile robot, Koala, and tested in various environments. Experimental results are presented which demonstrate the effectiveness of the proposed fuzzy navigation system. PMID- 16366248 TI - Monitoring and fault diagnosis of hybrid systems. AB - Many networked embedded sensing and control systems can be modeled as hybrid systems with interacting continuous and discrete dynamics. These systems present significant challenges for monitoring and diagnosis. Many existing model-based approaches focus on diagnostic reasoning assuming appropriate fault signatures have been generated. However, an important missing piece is the integration of model-based techniques with the acquisition and processing of sensor signals and the modeling of faults to support diagnostic reasoning. This paper addresses key modeling and computational problems at the interface between model-based diagnosis techniques and signature analysis to enable the efficient detection and isolation of incipient and abrupt faults in hybrid systems. A hybrid automata model that parameterizes abrupt and incipient faults is introduced. Based on this model, an approach for diagnoser design is presented. The paper also develops a novel mode estimation algorithm that uses model-based prediction to focus distributed processing signal algorithms. Finally, the paper describes a diagnostic system architecture that integrates the modeling, prediction, and diagnosis components. The implemented architecture is applied to fault diagnosis of a complex electro-mechanical machine, the Xerox DC265 printer, and the experimental results presented validate the approach. A number of design trade offs that were made to support implementation of the algorithms for online applications are also described. PMID- 16366249 TI - Wavelet-based combined signal filtering and prediction. AB - We survey a number of applications of the wavelet transform in time series prediction. We show how multiresolution prediction can capture short-range and long-term dependencies with only a few parameters to be estimated. We then develop a new multiresolution methodology for combined noise filtering and prediction, based on an approach which is similar to the Kalman filter. Based on considerable experimental assessment, we demonstrate the powerfulness of this methodology. PMID- 16366250 TI - An experimental bias-variance analysis of SVM ensembles based on resampling techniques. AB - Recently, bias-variance decomposition of error has been used as a tool to study the behavior of learning algorithms and to develop new ensemble methods well suited to the bias-variance characteristics of base learners. We propose methods and procedures, based on Domingo's unified bias-variance theory, to evaluate and quantitatively measure the bias-variance decomposition of error in ensembles of learning machines. We apply these methods to study and compare the bias-variance characteristics of single support vector machines (SVMs) and ensembles of SVMs based on resampling techniques, and their relationships with the cardinality of the training samples. In particular, we present an experimental bias-variance analysis of bagged and random aggregated ensembles of SVMs in order to verify their theoretical variance reduction properties. The experimental bias-variance analysis quantitatively characterizes the relationships between bagging and random aggregating, and explains the reasons why ensembles built on small subsamples of the data work with large databases. Our analysis also suggests new directions for research to improve on classical bagging. PMID- 16366251 TI - A hierarchical particle swarm optimizer and its adaptive variant. AB - A hierarchical version of the particle swarm optimization (PSO) metaheuristic is introduced in this paper. In the new method called H-PSO, the particles are arranged in a dynamic hierarchy that is used to define a neighborhood structure. Depending on the quality of their so-far best-found solution, the particles move up or down the hierarchy. This gives good particles that move up in the hierarchy a larger influence on the swarm. We introduce a variant of H-PSO, in which the shape of the hierarchy is dynamically adapted during the execution of the algorithm. Another variant is to assign different behavior to the individual particles with respect to their level in the hierarchy. H-PSO and its variants are tested on a commonly used set of optimization functions and are compared to PSO using different standard neighborhood schemes. PMID- 16366252 TI - An improved robust fuzzy-PID controller with optimal fuzzy reasoning. AB - Many fuzzy control schemes used in industrial practice today are based on some simplified fuzzy reasoning methods, which are simple but at the expense of losing robustness, missing fuzzy characteristics, and having inconsistent inference. The concept of optimal fuzzy reasoning is introduced in this paper to overcome these shortcomings. The main advantage is that an integration of the optimal fuzzy reasoning with a PID control structure will generate a new type of fuzzy-PID control schemes with inherent optimal-tuning features for both local optimal performance and global tracking robustness. This new fuzzy-PID controller is then analyzed quantitatively and compared with other existing fuzzy-PID control methods. Both analytical and numerical studies clearly show the improved robustness of the new fuzzy-PID controller. PMID- 16366253 TI - Pose estimation for augmented reality applications using genetic algorithm. AB - This paper describes a genetic algorithm that tackles the pose-estimation problem in computer vision. Our genetic algorithm can find the rotation and translation of an object accurately when the three-dimensional structure of the object is given. In our implementation, each chromosome encodes both the pose and the indexes to the selected point features of the object. Instead of only searching for the pose as in the existing work, our algorithm, at the same time, searches for a set containing the most reliable feature points in the process. This mismatch filtering strategy successfully makes the algorithm more robust under the presence of point mismatches and outliers in the images. Our algorithm has been tested with both synthetic and real data with good results. The accuracy of the recovered pose is compared to the existing algorithms. Our approach outperformed the Lowe's method and the other two genetic algorithms under the presence of point mismatches and outliers. In addition, it has been used to estimate the pose of a real object. It is shown that the proposed method is applicable to augmented reality applications. PMID- 16366254 TI - Bayesian model averaging of Bayesian network classifiers over multiple node orders: application to sparse datasets. AB - Bayesian model averaging (BMA) can resolve the overfitting problem by explicitly incorporating the model uncertainty into the analysis procedure. Hence, it can be used to improve the generalization performance of Bayesian network classifiers. Until now, BMA of Bayesian network classifiers has only been performed in some restricted forms, e.g., the model is averaged given a single node-order, because of its heavy computational burden. However, it can be hard to obtain a good node order when the available training dataset is sparse. To alleviate this problem, we propose BMA of Bayesian network classifiers over several distinct node-orders obtained using the Markov chain Monte Carlo sampling technique. The proposed method was examined using two synthetic problems and four real-life datasets. First, we show that the proposed method is especially effective when the given dataset is very sparse. The classification accuracy of averaging over multiple node-orders was higher in most cases than that achieved using a single node-order in our experiments. We also present experimental results for test datasets with unobserved variables, where the quality of the averaged node-order is more important. Through these experiments, we show that the difference in classification performance between the cases of multiple node-orders and single node-order is related to the level of noise, confirming the relative benefit of averaging over multiple node-orders for incomplete data. We conclude that BMA of Bayesian network classifiers over multiple node-orders has an apparent advantage when the given dataset is sparse and noisy, despite the method's heavy computational cost. PMID- 16366255 TI - Adaptive output feedback tracking control for a class of uncertain nonlinear systems using neural networks. AB - This correspondence addresses the problem of designing robust tracking control for a class of uncertain nonlinear MIMO second-order systems. An adaptive neural network-based output feedback tracking controller is constructed such that all the states and signals involved are uniformly bounded and the tracking error is uniformly ultimately bounded. Only the output measurement is required for feedback. The implementation of the neural network basis functions depends only on the desired reference trajectory. Therefore, the intelligent adaptive output feedback controller developed here possesses the properties of computational simplicity and easy implementation. A simulation example of controlling mass spring-damper mechanical systems is made to confirm the effectiveness and performance of the developed control scheme. PMID- 16366256 TI - Improved global robust asymptotic stability criteria for delayed cellular neural networks. AB - This paper considers the problem of global robust stability analysis of delayed cellular neural networks (DCNNs) with norm-bounded parameter uncertainties. In terms of a linear matrix inequality, a new sufficient condition ensuring a nominal DCNN to have a unique equilibrium point which is globally asymptotically stable is proposed. This condition is shown to be a generalization and improvement over some previous criteria. Based on the stability result, a robust stability condition is developed, which contains an existing robust stability result as a special case. An example is provided to demonstrate the reduced conservativeness of the proposed results. PMID- 16366257 TI - Stability analysis of fuzzy control systems subject to uncertain grades of membership. AB - This paper presents relaxed stability conditions for fuzzy control systems subject to parameter uncertainties. As the parameter uncertainties introduce uncertain grades of membership to the fuzzy control systems, the favorable property offered by sharing the same premises in the fuzzy plant models and fuzzy controllers cannot be employed to enhance the stabilization ability of the fuzzy control systems. To widen the applicability of the fuzzy control approach, fuzzy control systems subject to uncertain grades of membership will be investigated. New relaxed stability conditions will be derived to guarantee the stability of this class of fuzzy control systems. A numerical example will be given to show the effectiveness of the proposed approach. PMID- 16366259 TI - Modeling uncertainty in clinical diagnosis using fuzzy logic. AB - This paper describes a fuzzy approach to computer-aided medical diagnosis in a clinical context. It introduces a formal view of diagnosis in clinical settings and shows the relevance and possible uses of fuzzy cognitive maps. A constraint satisfaction method is introduced that uses the temporal uncertainty in symptom durations that may occur with particular diseases. The method results in an estimate of the stage of the disease if the temporal constraints of the disease in relation to the occurrence of the symptoms are satisfied. A lightweight fuzzy process is described and evaluated in the context of diagnosis of two confusable diseases. The process is based on the idea of an incremental simple additive model for fuzzy sets supporting and negating particular diseases. These are combined to produce an index of support for a particular disease. The process is developed to allow fuzzy symptom information on the intensity and duration of symptoms. Results are presented showing the effectiveness of the method for supporting differential diagnosis. PMID- 16366258 TI - From fuzzy logic toward plurimonism: the science of active and empathic observation. AB - Plurimonism is a new philosophy and method of science. It holds that the revolution in computer science and artificial intelligence has reached the point that all the sciences in general can now account for the complex relations of an irreducible plurality of unique observers engaged in describing the same event. Plurimonism seeks to describe the conscious and unconscious relations of the scientific observer during the act of observation of a given event while preserving the historical uniqueness and indivisible identity of each such observer. Using the framework of plurimonism, we mathematically formulate the problem of empathy. This self-reflective mathematical model entails four components of the empathic process involving two observers. They are: 1) the self; 2) the self's-other; 3) the other; and 4) the other's-self. It measures the degree of accuracy of the therapist-observer's empathy, as well as conscious and unconscious processes involved in the patient-observer's idealization and the therapist-observer's confidence in clinical psychotherapy. Ratings are obtained from both patient and therapist from four different points of view. The plural views of the patient's global assessment of functioning (GAF) are from: 1) the therapist's view (TGAF); 2) the patient's view (PGAF); 3) the therapist empathic view (TEGAF), which represents the therapist's estimate of PGAF; and 4) the patient's empathic estimate of the TGAF. The GAF scale is the standard dimensional 100-point-scale measure used in psychiatry for recording a patient's functioning. The patient's estimate of the therapist's degree of accuracy as well as the therapist's confidence in his or her empathic accuracy is also represented. Three formulae are presented that describe the degree of the therapist's empathic accuracy, the patient's over-idealization/under idealization, and the therapist's over-confidence/under-confidence. The concept of empathy is here restricted to mean the degree to which one observer can take the point of view of another observer when both are observing the same thing. PMID- 16366260 TI - Modeling gene expression networks using fuzzy logic. AB - Gene regulatory networks model regulation in living organisms. Fuzzy logic can effectively model gene regulation and interaction to accurately reflect the underlying biology. A new multiscale fuzzy clustering method allows genes to interact between regulatory pathways and across different conditions at different levels of detail. Fuzzy cluster centers can be used to quickly discover causal relationships between groups of coregulated genes. Fuzzy measures weight expert knowledge and help quantify uncertainty about the functions of genes using annotations and the gene ontology database to confirm some of the interactions. The method is illustrated using gene expression data from an experiment on carbohydrate metabolism in the model plant Arabidopsis thaliana. Key gene regulatory relationships were evaluated using information from the gene ontology database. A new regulatory relationship concerning trehalose regulation of carbohydrate metabolism was also discovered in the extracted network. PMID- 16366261 TI - Transcranial ultrasonography system for visualizing skull and brain surface aided by fuzzy expert system. AB - A conventional ultrasonography system can noninvasively provide human tissue and blood flow velocity information with real-time processing. In general, since the human skull prevents the disclosure of brain anatomy, we usually placed the sensor at the anterior and superior attachment site of the upper ear (the posterior temporal window) in adults. Due to this limitation, the conventional system cannot obtain transcranial information from arbitrary places in the skull. This paper describes a transcranial sonography system that can visualize the shape of the skull and brain surface from any point to examine skull fracture and brain disease such as cerebral atrophy and epidural or subdural hematoma. In this system, we develop anatomical knowledge of the human head, and we employ fuzzy inference to determine the skull and brain surface. To evaluate our method, three models are applied: the phantom model, the animal model with soft tissue, and the animal model with brain tissue. In all models, the shapes of the skull and the brain tissue surface are successfully determined. Next, the method is applied to two adults. As a result, we have determined the skin surface, skull surface, skull bottom, and brain tissue surface for the subjects' foreheads. Consequently, our system can provide the skull and brain surface information using three dimensional shapes. PMID- 16366262 TI - Modeling gunshot bruises in soft body armor with an adaptive fuzzy system. AB - Gunshots produce bruise patterns on persons who wear soft body armor when shot even though the armor stops the bullets. An adaptive fuzzy system modeled these bruise patterns based on the depth and width of the deformed armor given a projectile's mass and momentum. The fuzzy system used rules with sinc-shaped if part fuzzy sets and was robust against random rule pruning: Median and mean test errors remained low even after removing up to one fifth of the rules. Handguns shot different caliber bullets at armor that had a 10%-ordnance gelatin backing. The gelatin blocks were tissue simulants. The gunshot data tuned the additive fuzzy function approximator. The fuzzy system's conditional variance V[Y/X = x] described the second-order uncertainty of the function approximation. Handguns with different barrel lengths shot bullets over a fixed distance at armor-clad gelatin blocks that we made with Type 250 A Ordnance Gelatin. The bullet-armor experiments found that a bullet's weight and momentum correlated with the depth of its impact on armor-clad gelatin (R2 = 0.881 and p-value < 0.001 for the null hypothesis that the regression line had zero slope). Related experiments on plumber's putty showed that highspeed baseball impacts compared well to bullet armor impacts for large-caliber handguns. A baseball's momentum correlated with its impact depth in putty (R2 = 0.93 and p-value < 0.001). A bullet's momentum similarly correlated with its armor-impact in putty (R2 = 0.97 and p-value < 0.001). A Gujarati-Chow test showed that the two putty-impact regression lines had statistically indistinguishable slopes for p-value = 0.396. Baseball impact depths were comparable to bullet-armor impact depths: Getting shot with a .22 caliber bullet when wearing soft body armor resembles getting hit in the chest with a 40-mph baseball. Getting shot with a .45 caliber bullet resembles getting hit with a 90-mph baseball. PMID- 16366263 TI - Proteome analysis: a successful approach for functional research in recalcitrant non-model crops. PMID- 16366264 TI - Inhibition of SFO1, a putative E3 ubiquitin ligase from Arabidopsis, increases susceptibility to both host and non-host pathogenic fungi. PMID- 16366265 TI - Impact of non typical food matrice and cell density on Bacillus cereus emetic toxin production. PMID- 16366266 TI - Detection of N-acyl-L-homoserine lactones degradation by Bacillus spp. PMID- 16366267 TI - beta-Elimination of carrot pectin: towards a better understanding of carrot texture during thermal processing. PMID- 16366268 TI - Fire risk assessment in savanna ecosystems with multi-temporal satellite data. PMID- 16366269 TI - Soil moisture retrieval from optical and thermal spaceborne remote sensing. PMID- 16366270 TI - Optimal use of animal manure as a fertilizer. PMID- 16366271 TI - Detection of polymorphisms in the myostatin gene in Belgian Pietrain pigs. PMID- 16366272 TI - Milk odd and branched-chain fatty acids to predict ruminal methanogenesis in dairy cows. PMID- 16366273 TI - Heterologous production of human beta-defensin-2 in Arabidopsis thaliana. PMID- 16366274 TI - Simulating drift by CFD: sensitivity analysis of machine and environmental parameters. PMID- 16366275 TI - An optical tongue based on ATR-FTIR spectroscopy to taste tomatoes. PMID- 16366276 TI - Hyperspectral virtual imaging system of a Fagus sylvatica stand. PMID- 16366277 TI - Molecular engineering of an endoxylanase enzyme towards inhibitor insensitivity. PMID- 16366278 TI - Detection and isolation method for bacterial lysozyme inhibitors by use of lysozyme affinity chromatography. PMID- 16366279 TI - Isolation and partial characterisation of the protein fraction of brewers' spent grain. PMID- 16366280 TI - Cryopreservation and abiotic stress tolerance in potato: a proteomic approach. PMID- 16366281 TI - Antibiotic induced autolysis in Escherichia coli is independent of the level of lysozyme inhibitor ivy, but increases with plasmid insertion. PMID- 16366282 TI - The orotidine monophosphate decarboxylase gene of Myrothecium gramineum: cloning and sequence analysis. PMID- 16366283 TI - Evaluation of different methods for nucleic acids quantification. PMID- 16366284 TI - Production of L-ribulose by dehydrogenation of ribitol with Gluconobacter oxydans. PMID- 16366285 TI - Performance of silica filled poly (1-trimethylsilyl-1-propyne) nanocomposites. PMID- 16366286 TI - Measurement and modelling of water desorption isotherms for pasta at different temperatures. PMID- 16366287 TI - Dissection of the linker-2 intervening sequence in the transcriptional regulation of the Escherichia coli carab operon. PMID- 16366288 TI - Intergeneric crossings in Ranunculaceae: chromosome doubling of Ranunculus asiaticus. PMID- 16366289 TI - Development of ecological indicator methods based on macroinvertebrates for river quality assessment in Ecuador. PMID- 16366290 TI - Climate indices for Yazd region in Iran. PMID- 16366291 TI - High pressure mediated lysogenic conversion of Escherichia coli with Shiga-toxin encoding bacteriophage. PMID- 16366292 TI - Reflectance indices indicative for changes in water contents of chicory leaves. PMID- 16366293 TI - Model-based fuzzy controlled greenhouse cooling ventilation system. PMID- 16366294 TI - Decontamination of minimally processed vegetables by chlorine dioxide gas. PMID- 16366295 TI - Development of an atmospheric plasma process for the fabrication of biofunctional coatings. PMID- 16366296 TI - Peptidomics of Caenorhabditis elegans: in search of neuropeptides. PMID- 16366297 TI - Influence of ultrasonic waves during bleaching process on volatile compounds of Tunisian olive oil. PMID- 16366298 TI - Shape modelling of fruit by image processing. PMID- 16366299 TI - Eco-climatic image segmentation based on time series. PMID- 16366300 TI - Differences in extent of lipolysis in red or white clover and ryegrass silages in relation to polyphenol oxidase activity. PMID- 16366301 TI - Development and predation of Picromerus bidens (Heteroptera: Pentatomidae) on Spodoptera littoralis (Lepidoptera: Noctuidae) at different constant temperatures. PMID- 16366302 TI - Multiplicative scatter correction data linearization for on-line spectroscopy application. PMID- 16366303 TI - Virtual tissue generation of fruit tissue using Voronoi tessellations. PMID- 16366304 TI - Cu and Zn distribution in a constructed wetland for treatment of agricultural wastewater. PMID- 16366305 TI - Phytoextraction of Cd and Zn with Salix dasyclados Loden. PMID- 16366306 TI - Antimicrobial activity of selected phytochemicals against the pig gut flora. PMID- 16366307 TI - Quorum sensing dependent production of antimicrobial component influences establishment of E. coli in dual species biofilms with Serratia plymuthica. PMID- 16366308 TI - Development of a biological water quality assessment system for the Sumberjaya watershed in Sumatra (Indonesia). PMID- 16366309 TI - A pH dependent mechanism for inactivation of gram-negative bacteria by hen egg white and lambda lysozymes under high pressure. PMID- 16366310 TI - Predictive ecohydrological modelling using the random forest algorithm. PMID- 16366311 TI - A study of the L-ascorbate biosynthetic capacity of apple fruit. PMID- 16366312 TI - Framework and indicators for the quantification of erosion reduction by tree roots. PMID- 16366313 TI - The effect of headspace renewal in a temporary immersion bioreactor (TIB) on plantain shoot proliferation and quality. PMID- 16366314 TI - Comparison of taste and aroma of integrated and organic apple fruit. PMID- 16366315 TI - A general approach for recombinant protein expression in E. coli. PMID- 16366316 TI - Germination, growth and dry matter accumulation of wild almond seedlings from stratified and scarified seeds. PMID- 16366317 TI - Biological and conventional production of 14 varieties of durum wheat in the semi arid areas of Tunisia. PMID- 16366318 TI - Analysis of CO2 efflux rate and xylem CO2 partial pressure of young tree stems during the dormant season. PMID- 16366319 TI - Functional analysis of Arabidopsis thaliana plant defensins. PMID- 16366320 TI - Isobaric-isothermal degradation kinetics of (6R,S) 5-formyltetrahydrofolic acid. PMID- 16366321 TI - Enzymatic firming of high-pressure frozen strawberry halves by means of exogenous pectinmethylesterase. PMID- 16366322 TI - Humic fingerprinting of soil organic matter as a proxy for palaeo-reconstruction. PMID- 16366323 TI - Experimentally implanted populations of Hyacinthoides non-scripta: results after 45 years. PMID- 16366324 TI - A discrete element model for simulation of a spinning disc fertilizer spreader. PMID- 16366325 TI - Use of specific milk fatty acids as diagnostic tool for rumen acidosis in dairy cows. PMID- 16366326 TI - Agricultural policy and environment--need and impacts of agricultural support in India. PMID- 16366327 TI - EDC-mediated DNA attachment to nanocrystalline diamond. PMID- 16366329 TI - Predicting the occurrence of benthic species in the North Sea. PMID- 16366328 TI - High throughput enzymatic taste biosensor. PMID- 16366330 TI - Effective approaches to improve water use efficiency in dryland farming. PMID- 16366331 TI - Development of population models for Esox lucius and Tinca tinca in Flanders based on classification tree methods. PMID- 16366332 TI - Antioxidant enzyme activity during low temperature storage of potato tuber. PMID- 16366333 TI - Mechanism of severe water stress induced alterations in assimilate partitioning and transport in tomato plants. PMID- 16366334 TI - Hydroxyapatite reinforced poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3 hydroxyvalerate) based degradable composite bone plate. AB - Poly(3-hydroxybutyrate) (P3HB), its co-polymers with 3-hydroxyvalerate (HV) (PHBV8 and PHBV22), and their hydroxyapatite (HAp) containing composites (5 and 15%, w/w) were prepared by injection molding. PHBV bone plates with low valerate contents and 15% (w/w) HAp appear to have better mechanical properties than the others. Flexural strengths of 15% (w/w) HAp-loaded P3HB, PHBV8 and PHBV22 were 78.28, 63.45 and 39.38 MPa, respectively. Tensile strengths of 15% (w/w) HAp loaded P3HB, PHBV8 and PHBV22 were 18.99, 15.44 and 11.02 MPa, respectively. For the ageing test, bone plates were incubated in phosphate-buffered saline PBS (0.1 M, pH 7.4) at 37 degrees C and at pre-determined time points they were removed and subjected to a three-point bending test. Incubation in PBS caused a sharp decrease in the mechanical properties within the first 24 h, followed either by a gradual decrease or no change for a period of about 1 month. SEM results showed that there was no significant material erosion in the 4-week incubation period. P3HB loaded with 15% HAp appeared to yield the most suitable bone plate, insofar as mechanical properties are concerned with potential for further testing in vivo. PMID- 16366335 TI - Preparation and antibacterial test of chitosan/PAA/PEGDA bi-layer composite membranes. AB - Chitosan/poly(acrylic acid)/poly(ethylene glycol) diacrylate (PEGDA) composite membranes with a bi-layer configuration were prepared and their potential application as an antibacterial material was examined. A two-step process was adopted. A dope consisting of PEGDA, acrylic acid (AA) and a photoinitiator was cast and then UV-cured on a glass substrate to form a mechanically strong, dense membrane. Subsequently, the membrane was coated with a layer of solution composed of chitosan, AA and water. As the majority of AA diffused downwards into the supporting layer underneath, chitosan coagulated with residual AA to form a nano layer on the top surface by means of UV irradiation. Low-voltage field-emission scanning electron microscopy was used to observe the membrane morphology and to measure the thickness of the top layer. Contact angle measurements indicated a top layer mixed with chitosan and poly(acrylic acid), which was confirmed by chemical composition analysis of X-ray photon spectroscopy. The antibacterial activities of the formed membranes were tested both with respect to a Gram negative (Escherichia coli) and a Gram-positive (Staphylococcus aureus) bacteria. PMID- 16366336 TI - In vitro and in vivo degradation of non-woven materials made of poly(epsilon caprolactone) nanofibers prepared by electrospinning under different conditions. AB - The aim of this study was to prepare non-woven materials from a biodegradable polymer, poly(epsilon-caprolactone) (PCL) by electrospinning. PCL was synthesized by ring-opening polymerization of epsilon-caprolactone in bulk using stannous octoate as the catalyst under nitrogen atmosphere. PCL was then processed into non-woven matrices composed of nanofibers by electrospinning of the polymer from its solution using a high voltage power supply. The effects of PCL concentration, composition of the solvent (a mixture of chloroform and DMF with different DMF content), applied voltage and tip-collector distance on fiber diameter and morphology were investigated. The diameter of fibers increased with the increase in the polymer concentration and decrease in the DMF content significantly. Applied voltage and tip-collector distance were found critical to control 'bead' formation. Elongation-at-break, ultimate strength and Young's modulus were obtained from the mechanical tests, which were all increased by increasing fiber diameter. The fiber diameter significantly influenced both in vitro degradation (performed in Ringer solution) and in vivo biodegradation (conducted in rats) rates. In vivo degradation was found to be faster than in vitro. Electrospun membranes were more hydrophobic than PCL solvent-casted ones; therefore, their degradation was a much slower process. PMID- 16366337 TI - Core-shell microspheres by dispersion polymerization as promising delivery systems for proteins. AB - Functional poly(methyl methacrylate) core-shell microspheres were prepared by dispersion polymerization. An appropriate selection of experimental parameters and in particular of the initiator and stabilizer amount and of the medium solvency power allowed a monodisperse sample as large as 600 nm to be prepared. To this purpose, low initiator concentration, high steric stabilizer amount and a low solvency power medium were employed. The microspheres present a core-shell structure in which the outer shell is constituted by the steric stabilizer which affords carboxylic groups able to interact with basic proteins, such as trypsin, whose adsorption is essentially driven by the carboxylic group density in the microsphere shell. Finally, fluorescent microspheres were prepared for biodistribution studies and shown to be readily taken up by the cells both in vitro and in vivo. These results suggest that these microspheres are promising delivery systems for the development of novel protein-based vaccines. PMID- 16366338 TI - Chemical modification of chitosan by phosphorylation: an XPS, FT-IR and SEM study. AB - In the present work, the surface of chitosan membranes was modified using a phosphorylation method carried out at room temperature. Phosphorylation may be of particular interest in materials for orthopaedic applications, due to the cation exchange properties of phosphate functionalities. Phosphate groups chelate calcium ions, thus inducing the deposition of an apatite-like layer known to improve the osteoconduction of polymer-based implants. Additionally, the negatively charged phosphate functionalities, together with the positively charged amine groups from chitosan, are expected to provide chitosan with an amphoteric character, which may be useful as a combinatorial therapeutic strategy, by simultaneously allowing the immobilization of signalling molecules like growth factors. Phosphorylation was carried out at room temperature using the H3PO4/Et3PO4/P2O5/butanol method. Surface characterization was performed by XPS, ATR-FT-IR, and SEM. Cross-sections were analyzed by SEM fitted with EDS. The phosphate content increased with the reaction time, as shown by XPS and ATR-FT IR, a P/N atomic ratio of 0.73 being obtained after 48 h of treatment. High resolution XPS spectra regarding C1s, O1s, N1s and P2p are discussed. The introduction of a neutralization step led to a reduction of P content, which pointed out to the presence of phosphates ionically bound to protonated amines, in addition to phosphate esters. EDS analysis of cross-sections revealed a gradual P reduction up to 50% towards the inner part of the membrane. PMID- 16366339 TI - Fabrication using a rapid prototyping system and in vitro characterization of PEG PCL-PLA scaffolds for tissue engineering. AB - In the field of tissue engineering new polymers are needed to fabricate scaffolds with specific properties depending on the targeted tissue. This work aimed at designing and developing a 3D scaffold with variable mechanical strength, fully interconnected porous network, controllable hydrophilicity and degradability. For this, a desktop-robot-based melt-extrusion rapid prototyping technique was applied to a novel tri-block co-polymer, namely poly(ethylene glycol)-block poly(epsilon-caprolactone)-block-poly(DL-lactide), PEG-PCL-P(DL)LA. This co polymer was melted by electrical heating and directly extruded out using computer controlled rapid prototyping by means of compressed purified air to build porous scaffolds. Various lay-down patterns (0/30/60/90/120/150 degrees, 0/45/90/135 degrees, 0/60/120 degrees and 0/90 degrees) were produced by using appropriate positioning of the robotic control system. Scanning electron microscopy and micro computed tomography were used to show that 3D scaffold architectures were honeycomb-like with completely interconnected and controlled channel characteristics. Compression tests were performed and the data obtained agreed well with the typical behavior of a porous material undergoing deformation. Preliminary cell response to the as-fabricated scaffolds has been studied with primary human fibroblasts. The results demonstrated the suitability of the process and the cell biocompatibility of the polymer, two important properties among the many required for effective clinical use and efficient tissue engineering scaffolding. PMID- 16366340 TI - Poly(ethylene glycol) hydrogels cross-linked by hydrolyzable polyrotaxane containing hydroxyapatite particles as scaffolds for bone regeneration. AB - Poly(ethylene glycol) (PEG) hydrogels cross-linked by a hydrolyzable polyrotaxane containing hydroxyapatite particles (PRX-HAp) were developed as scaffolds for bone regeneration. Five scaffolds with various composition of the polyrotaxane, PEG and HAp particles were prepared to examine cell adhesion in vitro using rat primary cultured osteoblast. Cells were observed to attach well on a PRX-HAp that have the same weight ratio of the polyrotaxane and HAp particles at 7 days after seeding. These results indicate that HAp particles are necessary for cell adhesion and survival, but a higher ratio of the particles is not suitable for cell adhesion. The composites of rat osteoblast and the PRX-HAp were implanted subcutaneously in syngeneic rats and harvested at 5 weeks after implantation. In histological analysis, osteoblast-like cells became arrayed along the surface of the PRX-HAp, and osteoid-like tissues were observed in the region between a queue of osteoblast-like cells and PRX-HAp. These images are similar to intramembranous ossification, and it is expected that bone regeneration occurs on the surface of the PRX-HAp. This study strongly suggests the great potential of the PRX-HAp as scaffolds for bone regeneration. PMID- 16366341 TI - Medical care for the Nigerian school child. PMID- 16366342 TI - Medical laboratory services in western Nigeria. PMID- 16366343 TI - The control of malaria in a holoendemic region. PMID- 16366344 TI - Africa, abortion and the law. PMID- 16366345 TI - Observations on bone marrow and peripheral leucocyte cultures of Burkitt's lymphoma patients: a preliminary report. PMID- 16366346 TI - Colon by-pass operation for chronic oesophageal obstruction. PMID- 16366347 TI - Human skin lesions in East Central State, Nigeria due to the larvae of Amblyomma variegatum (Fabricius, 1794). PMID- 16366348 TI - Complications and prognostic factors in pneumonia among Nigerians. PMID- 16366349 TI - Burns of the feet. PMID- 16366350 TI - Treatment of psychiatric illness in a general hospital in Nigeria. PMID- 16366351 TI - Clinical evaluation of moduretic in the treatment of arterial hypertension. PMID- 16366352 TI - Clinical trial of rifampicin in the treatment of pulmonary tuberculosis in Lagos. PMID- 16366353 TI - A case of acute appendicitis in pregnancy. PMID- 16366354 TI - Locked twins. PMID- 16366355 TI - Lassa fever: a new appraisal. PMID- 16366356 TI - [Respiratory syncytial virus infection]. AB - Human respiratory syncytial virus (RSV) is the most common worldwide cause of lower respiratory tract infections (LRI) in infants less than 12 months of age. RSV isolates can be divided into group A and B. In addition, there were many genotypes within each group, and these genotypes have evolved global setting with temporal and geographic clustering. Many cellular genes encoding cytokines and chemokines which are activated by RSV infection has now been focused for the elucidation of pathophysiology of RSV LRI. The prophylaxis against RSV infection by vaccination has been unsuccessful because of its adverse effects. No valuable anti-RSV drugs for clinical use have been yet developed. Therefore RSV LRI has been treated mainly symptomatically. Recently humanized anti-RSV F protein monoclonal antibody was developed and prescribed for prevention in high-risk infants such as premature ones and those with chronic lung and congenital heart diseases. It reduced the incidence of hospitalization significantly. PMID- 16366357 TI - [Symptoms of food-borne diseases and gastroenteritis in Kyushu, Japan]. AB - In this study we analyzed the symptoms of gastroenteritis or food-borne disease caused by the 10 most prevalent pathogens: Norovirus, Salmonella, Vibrio parahaemolyticus, Campylobacter jejuni, Clostridium perfringens, Shiga toxin producing Escherichia coli (STEC), enterotoxigenic E. coli (ETEC), Shigella sonnei/flexneri (Shigella), Staphylococcus aureus, and emetic-type Bacillus cereus. The symptoms diarrhea, vomiting, fever, abdominal pain, and headache, and the incubation period in 646 cases in 10 districts of Kyushu between January 2000 and December 2004 were recorded. The pathogen with the shortest mean incubation period was B. cereus (0.8 h), and was followed by S. aureus (3.3 h), C. perfringens (10.7 h) and V. parahaemolyticus (16.4 h). All the patients infected with B. cereus and S. aureus developed symptoms within 6 hours, and those infected with V. parahaemolyticus and C. perfringens developed symptoms within 24 hours. Bloody diarrhea was associated with STEC and Shigella, but rare with other pathogens. Vomiting was associated with almost all cases of S. aureus and B. cereus infection, and occurred in 71.5% of the Norovirus cases and 56.1% of the V. parahaemolyticus cases. Vomiting was less common in the C. perfringens (22.0%) and the ETEC and STEC (both about 5%). Bloody diarrhea, abdominal pain, and vomiting were statistically significantly more common with STEC 0157 infection than with STEC non-0157 infection. Since the cases analyzed in this study included all degrees of illness, mild to severe, and a wide range of ages, the information obtained will serve as a good reference material for administrative and laboratory work when an outbreak takes place. PMID- 16366358 TI - [Annual changes in antimicrobial susceptibility and macrolide resistance of Streptococcus pyogenes from 1995 through 2004]. AB - This aim of this study was to reveal annual changes in antibiotic susceptibility, especially the macrolide susceptibility of Streptococcus pyogenes. A total of 755 strains of S. pyogenes were clinicaly isolated from throat swabs of children from 1995 through 2004 in Chiba Municipal Kaihin Hospital. All isolates were fully susceptible to benzylpenicillin, cefotaxim and cefaclor. The rate of resistance to erythromycin (EM) was over 10% every year after 2001 and 19% in 2004, and the rate of high resistance (MIC > or =16 microg/mL) has been increasing. A significant increase in EM resistance was observed over a 10-year period. There were 118 strains (15.6%) that persisted after treatment with beta-lactams. In the past few years it has been discovered that some S. pyogenes can be internalized by human cells of respiratory tract origin and survive within them. Since beta lactams do not reach high intracellular concentrations, this ability of S. pyogenes may be related to treatment failure. Since macrolides can enter eukaryotic cells and remain active in intracellular compartments, they will be effective for these S. pyogenes. In case of pharyngitis which againist treatment with beta-lactams, there is a possibility macrolides are effective. Macrolides may be effective in pharyngitis resistant to treatment with beta-lactams. However, macrolide resistance is not rare, susceptibility must be tested. PMID- 16366359 TI - [Eleven cases of co-infection with influenza type A and type B suspected by use of a rapid diagnostic kit and confirmed by RT-PCR and virus isolation]. AB - In the 2004/05 influenza season there were epidemics of influenza caused by several types of viruses (type B and A (H3) viruses, and type B, A (H3), and A (H1) viruses) in many areas of Japan. In such epidemics a single individual could be co-infected with several influenza viruses. From February to March in 2005, we examined 15 patients who were positive for influenza type A and B viruses when tested with a rapid diagnostic kit. The type A (H3) and B influenza virus genes were successfully amplified by RT-PCR in 10 of the 15 patients, confirming that they were co-infected with type A (H3) and B viruses. The type A (H1) and B virus genes were successfully amplified in another patient, confirming that the patient was co-infected with type A (H1) and B viruses. By contrast, 2 patients were clearly positive for type A and B viruses according to the rapid diagnostic kit, but positive for type B virus alone by RT-PCR. No influenza virus genes were detected by RT-PCR in the remaining 2 patients. To isolate one type from a mixture of two different types of influenza viruses in a specimen, we neutralized one of the types with type-specific antiserum, and isolated the other with MDCK (+) cells. The results obtained by virus isolation were identical to those obtained by RT-PCR. Influenza viruses corresponding to the results of RT-PCR were isolated from 9 of the 11 patients in which isolation was attempted. No viruses were isolated from the 2 patients in whom no virus genes were detectable by RT PCR. Based on these results we concluded that 11 of 15 patients who were positive for type A and B viruses according to the rapid diagnostic kit were co-infected with type A (H3) or A (H1) and B virus. When several types of influenza viruses are prevalent, as in the 2004/05 influenza season, the possibility of a patient being co-infected with more than one type of influenza virus should be considered. PMID- 16366360 TI - [Investigation of Streptococcus pneumoniae and Haemophilus influenzae isolated from pediatric outpatients nationwide with a respiratory tract infection at the first consultation (2002-2003)--proportion of resistant strains and sensitivity to oral antibacterial agents]. AB - The propotions of resistant strains and sensitivity to oral antibacterial agents were determined for 468 strains of Streptococcus pneumoniae and 557 strains of Haemophilus influenzae isolated from pediatric outpatients with a respiratory tract infection at the first consultation at 20 medical institutions nationwide in the predefined 4-week period during November 2002 and June 2003. PRSP of the S. pneumoniae strains, accounted for 27%, and BLNAR accounted for 35% of the H. influenzae strains. Against the S. pneumoniae strains, faropenem showed the best MIC90 value, 0.5 microg/mL, of the beta-lactam antibacterial agents, followed by cefditoren-pivoxil, and among the macrolide and ketolide antibacterial agents, telithromycin showed the best MIC90 value, 0.12 microg/mL. Against the H. influenzae strains, cefditoren-pivoxil showed the best MIC90 value, 0.25 microg/mL, among the beta-lactam antibacterial agents, and among the macrolide and ketolide antibacterial agents, azithromycin showed the best activity, followed by telithromycin. It seems necessary to take these results into consideration when choosing drugs for treatment. It will be necessary to survey the trends in resistant strains and the clinical efficacy of various oral antibacterial agents in the future. PMID- 16366361 TI - [Yersinia pseudotuberculosis type 4a infection meeting the diagnostic criteria for Kawasaki disease complicated by disseminated intravascular coagulation]. AB - We report a case of Yersinia pseudotuberculosis (Y. ptbc) infection complicated by disseminated intravascular coagulation (DIC) that presented as Kawasaki disease (KD). A 9-year-old girl had been well until two days before, when she developed a fever, exanthem, and abdominal pain. An erythematous macular rash was observed in the perineum, and she had a strawberry tongue. The patient was admitted to Kawasaki Medical School Hospital because the macular rash spread over her entire body, and edema of her hands and conjunctivitis subsequently developed. Echo cardiography showed dilation of the left coronary artery. Thrombocytopenia and an elevated total fibrin degeneration product level were noted on the third hospital day, and the prothronmbin and partial-thromboplastin times were prolonged. Her clinical presentation was typical of KD and DIC. A stool culture and a blood culture were negative. Serologic tests were positive for antibodies to Y. ptbc. The antibody titer against Y. ptbc-derived mitogen was not elevated after her recovery. Y. ptbc infection should be considered in an older child whose clinical findings fulfill the criteria for KD complicated by DIC. PMID- 16366362 TI - [Clinical analysis of home oxygen therapy in patients with pneumoconiosis at Iwamizawa Rosai Hospital]. AB - Although home oxygen therapy (HOT) has become one of the standard therapies in the management of chronic respiratory failure, especially for patient with hypoxia, there are very few studies about HOT in patients with pneumoconiosis. Therefore, we clinically investigated the characteristics of 97 patients with pneumoconiosis and 30 patients with COPD on home oxygen therapy (HOT) followed at Iwamizawa Rosai Hospital during the period from 1990 to 2003. The mean survival time in patients with pneumoconiosis (42 months) by the Kaplan-Meier method was shorter than in patents with COPD (75 months). Among the patients with pneumoconiosis divided by chest radiography classification of pneumoconiosis, there was no significant difference in mean survival time. The survival time in pneumoconiosis patients with dyspnea grade IV was shorter than in patients with dyspnea grade III. PMID- 16366363 TI - [Clinical and pathological findings of patients with interstitial lung disease associated with antisynthetase]. AB - Autoantibodies against aminoacyl-tRNA synthetase (ARS) are known to associate with myositis, arthritis, and interstitial lung disease (ILD). Anti Jo-1 antibody is a common diagnostic tool of this syndrome and is available on a commercial basis, while other anti-ARS antibodies such as anti-EJ, anti-PL7 and anti-OJ antibodies are less commonly examined. The aim of this study was to investigate the clinicopathological characteristics of 9 ILD patients who were positive for anti-ARS antibodies including anti-EJ, anti-PL7 and anti-Jo-1 antibodies. In physical examination, muscle pain and/or skin rash (including mechanic's hand) were observed frequently. In laboratory findings, levels of myogenic enzymes were slightly elevated, and anti-nucleus antibody was detected in most cases. Air space consolidation with traction bronchiectasis, ground glass opacities and characteristic volume loss of both lower lobes were observed in radiological examination. Organization in alveolar spaces and lymphoid alveolitis that did not destroy existing pulmonary structures were main pathological findings. Immunosupressive therapy was effective and the prognosis appears satisfactory in these cases. Although anti-ARS antibodies had been measured as a marker of myositis, we suggest that they are useful as markers of interstitial pneumonia. PMID- 16366364 TI - [A case of lung cancer with cranial neuropathy as the first sign of onset due to metastatic leptomeningeal carcinomatosis]. AB - A 68-year-old man suffered right facial palsy and left deafness, however, his condition was considered to be idiopathic and he was followed. Three months later, bloody sputum and hoarseness caused him to be admitted to our hospital. An abnormal shadow was detected in the right upper lung field and adenocarcinoma of the lung with multiple brain metastases was diagnosed. He underwent gamma-knife radiosurgery for the brain lesions and subsequent systemic chemotherapy consisting of combined carboplatin and paclitaxel, which were not effective. Subsequently various neurological symptoms appeared, such as muscle weakness of the extremities, dizziness, and gait disturbance. Adenocarcinoma cells confirmed in the cerebrospinal fluid were similar to those in the obtained by transbronchial curetting. Whole-brain irradiation was performed, however, the neurological symptoms worsened and he died. Leptomeningeal carcinomatosis is difficult to diagnose while the patient is alive. It is thought that cranial neuropathy due to leptomeningeal carcinomatosis is a rare form of onset for lung cancer. PMID- 16366365 TI - [A case of Clostridium difficile colitis associated with paclitaxel and carboplatin chemotherapy for lung cancer]. AB - A 41-year-old man admitted complaining of dyspnea was found to have lung adenocarcinoma (T4N3M1, stage IV) originating from S1+2. He underwent chemotherapy with carboplatin (CBDCA) and paclitaxel (PTX) and partial remission was obtained. However, on the 11th day of the fourth course of chemotherapy, he developed lower abdominal pain, grade 3 bloody diarrhea and grade 2 vomiting. The stool contained Clostridium difficile (CD) toxin and stool culture revealed C. difficile growth. We diagnosed CD colitis. Fortunately his symptoms recovered by fasting and fluid replacement until the next day. It has been reported that CD colitis occurrs in approximately 2% of patients with gynecological cancers receiving PTX-including chemotherapy. We thus speculate that the CD colitis of the present case was due to PTX. Although the mechanism of CD colitis by chemotherapeutical agents remains undetermined, direct damage to intestinal mucosa or changes in intestinal bacterial flora are possible. As severe CD colitis can be life threatening, we have to be aware of the possibility of it occurring in patients undergoing chemotherapy. PMID- 16366366 TI - [An adult case of asymptomatic congenital tracheal stenosis]. AB - A 42-year-old woman was admitted with abnormal chest radiographs. Though interstitial pneumonia associated with dermatomyositis was diagnosed, her chest radiograph also revealed a narrowed trachea about 6 mm in diameter. Bronchoscopy showed that her trachea lacked a membranous posterior segment and O-shaped complete tracheal rings were present throughout the trachea, indicating congenital tracheal stenosis. Congenital tracheal stenosis is a rare disorder and is usually recognized in the first few weeks of life, but the patient had no history of dyspnea or recurrent pneumonia. This case suggests that among healthy people there are a very few who have asymptomatic congenital tracheal stenosis. PMID- 16366367 TI - [Legionnaires' disease with acute renal failure caused by Legionella pneumophilla serogroup 4]. AB - A 77-year-old man who had fever and chest pain was admitted to a neighboring hospital on a diagnosis of pneumonia. Chest X-ray film finding deteriorated despite treatment with 2 g cefotaxime per day. Because of accompanying acute renal failure, he was transferred to our hospital. Hemodialysis with intravenous administration of erythromycin and meropenem resulted in recovery from acute renal failure, and his general condition improved. Because of liver dysfunction, erythromycin was changed to pazufloxacin. Although he was negative for Legionella urinary antigen determined with a rapid assay kit, Binax NOW, his serum titer for Legionella pneumophila serogroup 4 was elevated. Finally, a diagnosis of Legionnaires' disease caused by Legionella pneumophila serogroup 4 was established. PMID- 16366368 TI - [Familial summer-type hypersensitivity pneumonitis--case report and review of literature]. AB - A 37-year-old-woman was admitted to our hospital because of chest bilateral reticular shadow with fever, cough, general malaise and exertional dyspnea in the summer. A diagnosis of summer-type hypersensitivity pneumonitis (SHP) was made by radiological, serological and histological examinations. Her 10-year-old daughter had chest reticular shadows and similar symptoms. These two patients were positive for serum anti-Trichosporon cutaneum (T. asahii, T. mucoides) antibodies and T. asahii was identified from cultured samples obtained from their house. They recovered spontaneously after hospitalization or isolation from the antigen. We reviewed the clinical features in sixteen families with familial SHP reported in Japan. Children aged under 15 years old accounted for 34% and there was no gender difference among patients. This finding differs from the conventionally defined features of patients with SHP. Measurements of serum KL-6, SP-D and SP-A seem to be useful for auxiliary diagnosis and monitoring the disease activity of SHP, especially in pediatric cases who cannot undergo invasive evaluation. PMID- 16366369 TI - [A case of hypersensitivity pneumonitis caused by inhalation of Mycobacterium avium from a home bath with a circulating water system]. AB - A 26-year-old man presented with complaints of exertional dyspnea and cough. The patient has already been given corticosteroids at a previous hospital. Chest CT revealed small centrilobular nodules with diffuse ground-glass opacities in both lungs. Lung biopsy specimens at thoracoscopy revealed non-necrotizing granulomas, patchy foci of mononuclear cell infiltration and fibrous thickening of alveolar septa, and Masson's bodies in bronchioles. Sputum culture showed the growth of Mycobacterium avium complex (MAC). Culture of water from the bath tub of his home showed MAC. Administration of antituberculous drugs and corticosteroids, and avoidance of bathing at home resulted in the improvement of his symptoms and CT findings. We believe the case is hypersensitivity pneumonitis to MAC in an immunocompetent patient, simulating hot tub lung. Hypersensitivity pneumonitis caused by MAC is rare in Japan. PMID- 16366370 TI - [A familial case of summer-type hypersensitivity pneumonitis possibly associated with bird breeder's lung diagnosed by bronchoalveolar lavage fluid]. AB - Case 1: A 32-year-old woman had cough and exertional dyspnea in August 2002, and chest computed tomographic scan revealed diffuse centrilobular nodules. Bronchoalveolar lavage fluid (BALF) showed a high proportion of lymphocytes with a decreased CD 4/CD 8 ratio. Transbronchial lung biopsy (TBLB) specimens showed alveolitis. Summer-type hypersensitivity pneumonitis was diagnosed on the basis of positive findings of anti-Trichosporon antibodies in the serum. Case 2: A 64 year-old man, the father of Case 1, also had cough and exertional dyspnea in August 2003. He had been in close contact with pigeons. Chest computed tomographic scan revealed bilateral map-like ground-glass opacities predominantly in the upper lobes. BALF showed a high proportion of lymphocytes with a decreased CD 4/CD 8 ratio. TBLB specimens showed alveolitis, granuloma and Masson body in the air spaces. Specific IgG and IgA antibodies against Trichosporon asahii, IgA antibodies against Trichosporon mucoides, and IgA antibodies against pigeon dropping extracts were found only in the BALF but not in the serum. Although a positive finding of returning-home provocation test was definitive in diagnosing summer-type hypersensitivity pneumonitis, he was also suspected of having bird fancier's lung. PMID- 16366371 TI - [A case of lung adenocarcinoma of the lung with disappearance of brain metastasis by re-treatment with gefitinib]. AB - BACKGROUND: Tumor response rate to gefitinib by previously treated patients with advanced non-small-cell lung cancer was approximately 20%. However, there are few reports about effect of re-treatment with gefitinib. CASE: A 40-year-old man was given a diagnosis of adenocarcinoma of lung (c-T2N3M1). Response was not obtained with chemotherapy (paclitaxel + carboplatin, vinorelbine + gencitabin). He developed brain metastasis and received whole brain radiation therapy. He was then given gefitinib. Reduction of the primary tumor, brain metastasis, pulmonary metastasis and liver metastasis was seen. Recurrence of pulmonary metastasis and liver metastasis was discovered 8 months after treatment with gefitinib. Therefore, treatment by gefitinib was stopped. Response was not achieved with chemotherapy (docetaxel and amrubicin). At the request of the patient, retreatment with gefitinib was resumed. Disappearance of brain metastasis was achieved. However, primary tumor, pulmonary metastasis and liver metastasis progressed. CONCLUSION: We reported a case whose brain metastases disappeared with re-treatment with gefitinib. This report is thought to be useful in terms of information about the acquired resistance of gefitinib. Further investigations are needed to determine the indication of re-treatment with gefitinib. PMID- 16366372 TI - Tumor ablation: treatment and palliation using image-guided therapy. PMID- 16366373 TI - Percutaneous ablation of kidney tumors in nonsurgical candidates. AB - Although resection currently remains the standard of care for renal carcinoma, the search for less invasive treatments has led to alternative surgical approaches. Even less invasive, and appropriate for many groups of patients, is percutaneous radiofrequency ablation, which induces tumor necrosis via lethal hyperthermia. Multiple series of renal tumors treated with percutaneous ablation in vivo and left in situ have been published; these series reveal that for small renal tumors, radiofrequency ablation results in complete necrosis at imaging in 79% to 100% of cases. Because current results come from tumors left in situ with short postablation follow-up, long-term results are necessary to compare outcomes to surgical standards. Complication rates are lower than those following partial nephrectomy. Future reports will shed light on the long-term outcomes of percutaneous ablation and the relative advantages and disadvantages of various technologies for thermal ablation. PMID- 16366374 TI - Radiofrequency ablation in lung cancer: promising results in safety and efficacy. AB - Only about 15% of patients diagnosed with lung carcinoma each year are surgical candidates, either due to advanced disease or comorbidities. The past decade has seen the emergence of minimally invasive therapies using thermal energy sources: radiofrequency, cryoablation, focused ultrasound, laser, and microwave; radiofrequency ablation (RFA) is the best developed of these. Radiofrequency ablation is safe and technically highly successful in terms of initial ablation. Long-term local control or complete necrosis rates drop considerably when tumors are larger than 3 cm, although repeat ablations can be performed. Patients with lung metastases tend to fare better with RF lung ablation than those with primary lung carcinoma in terms of local control, but it is unclear if this is related to smaller tumor size at time of treatment, lesion size uniformity, and sphericity with lung metastases, or to differences in patterns of pathologic spread of disease. The effects of RFA on quality of life, particularly dyspnea and pain, as well as long-term outcome studies are generally lacking. Even so, the results regarding RF lung ablation are comparable to other therapies currently available, particularlyfor the conventionally unresectable or high-risk lung cancer population. With refinements in technology, patient selection, clinical applications, and methods of follow-up, RFA will continue to flourish as a potentially viable stand-alone or complementary therapy for both primary and secondary lung malignancies in standard and high-risk populations. PMID- 16366375 TI - Percutaneous ablation: safe, effective treatment of bone tumors. AB - Percutaneous radiofrequency ablation (RFA) of osteoid osteomas has replaced surgical excision as the preferred method for treatment of these benign lesions, due to high effectiveness and low morbidity. Both RFA and cryoablation are safe and effective for palliation of pain due to metastatic disease in patients who have failed conventional therapies. These image-guided treatments can be performed precisely, allowing safe treatment of complex metastatic tumors. A single ablation treatment is effective in most patients, is well tolerated, and provides a long duration of pain relief PMID- 16366376 TI - Expanded treatment of hepatic tumors with radiofrequency ablation and cryoablation. AB - Three definitive treatment options are available for patients with hepatic tumors: hepatic resection, tumor ablation, and hepatic transplantation. Ablative techniques--including radiofrequency ablation (RFA) and cryoablation--have greatly improved physicians' ability to definitively treat patients with primary and secondary hepatic tumors. Both RFA and cryoablation are safe and well tolerated, but the effectiveness for local tumor eradication depends on many factors, including tumor size, location, number, and type. The choice of ablation modality is based on user and institutional biases. Assessing outcome after ablation is difficult because few studies with good long-term follow-up have evaluated local recurrence, disease-free survival, and overall survival after ablation. This and other limitations make it difficult to draw meaningful conclusions. PMID- 16366377 TI - [Experimental study on simultaneous selective reinnervation of the adductors and the abductor muscle for the treatment of the laryngeal paralysis]. AB - OBJECTIVE: The aim of this study is to estimate the value of a new surgical procedure in the treatment of the chronic unilateral laryngeal paralysis. METHODS: The recurrent laryngeal nerve of the left side of the dog was totally cut and served as a model of unilateral laryngeal paralysis at the first step of the research. The adductor and abductor branches of the recurrent laryngeal nerve were then, selected and cut. Afterwards, they were micro-sutured respectively with one branch of ansa cervicalis and phrenic nerve immediately (group 1) and 4 months later (group 2). Six months after this reinnervation, the laryngeal physiologic function of the lateral crico-arytenoid muscle (LCA) and the posterior crico-arytenoid muscle (PCA) have been checked by the methods of electromyography (EMG) and direct laryngoscopy. All the data have been analysed by the statistic methods. RESULTS: Among all the data of EMG, only the wave amplitude of action potential of the LCA muscle of the group 2 was diminished (p < 0.05). Under the direct laryngoscopy, the adductor movement of the left vocal cord of the group 2 was also lightly reduced. But the adductor and abductor movements of the left vocal cord were synchronous with the mouvements of the right vocal cord. CONCLUSION: Though the result of nervous reinnervation of a four month's laryngeal paralysis was not so good by comparison with that of an immediate reinnervation, this surgical procedure can however on the clinical point of view, reach a satisfactory level. The duration maximum of the reinnervation operation after laryngeal paralysis, is, at the present, not clear. It is necessary for us to make further studies. PMID- 16366378 TI - Polymorphisms of the human OGG1 gene in laryngeal cancer: implications in radiotherapy response and survival. AB - The human OGG1 (hOGG1) gene encodes a DNA glycosylase involved in the excision repair of 8-hydroxy-2'-deoxyguanine (8-OH-dG) from oxidatively-damaged DNA. Ser326Cys polymorphism in the hOGG1 gene is involved in the repair of 8 hydroxyguanine in oxidatively damaged DNA, and appears to be related to susceptibility to certain smoking and alcohol-related orolaryngeal cancers. OBJECTIVE: To analyse if hOGG1 Ser326Cys (exon 7: m6) polymorphism is associated with tumour localization, T, stage and histologic differentiation, and if radiotherapy results were influenced by this polymorphism. MATERIAL AND METHOD: Blood samples were obtained before treatment from seventy one patients with laryngeal cancer and screened by a PCR-RFLP method. RESULTS: Although hOGG1 gene is important in DNA repair mechanisms, no significant association was observed between hOGG1 Ser326Cys (exon 7: m6) polymorphism, tumour characteristics and radiotherapy results. CONCLUSION: So the analysis of this polymorphism is not important for treatment decision in laryngeal cancer patients. PMID- 16366379 TI - [Reconstruction after tumour's excision in lip's cancer: report of 100 cases]. AB - OBJECTIVE: Cancer of the lips is a frequent pathology, it represents 1/5 of cancers of the upper aerodigestive tract. Its objectives are to present the experience of the plastic and reconstructive surgery unit of the universitary hospital of Casablanca. MATERIAL AND METHOD: The authors present a retrospective study about 100 cases of lips cancer treated between January 1999 and December 2002. RESULTS: The average age is 59 years. The sex-ratio is 7.3. Eighty-five percent of our patients were smokers and 22% were alcoholics, the association alcoholic and smoker was found in 20%. In our study 71% were classified T1-T2 and 29% as T3-T4. The tumoral location was the lower lip in 82% and the upper lip in 10%. The histological study showed a squamous cell carcinoma type in 85% and a basal-cell carcinoma type in 14% of cases. After the tumour's excision, the reparation was done by cross-lip flap in 25%, the naso-lateral flap in 16% and the Gillies flap in 8%. The functional result was good in 92% of cases and the aesthetic result was excellent in 60% of cases and good in 30% of cases. The survival rate at 2 year was 90%. CONCLUSION: Actually, lip's cancer treatment is essentially surgical. The reconstructive techniques are various, depending on the size and location of the loss of substance. PMID- 16366380 TI - [Endobronchial lipomas and their treatment: one case report]. AB - PURPOSE: Starting from a clinical observation of an endobronchial lipoma and in the light of worldwide literature, the aim of this case report is to specify the epidemiology, the key points of the diagnosis and the advantage of endoscopy both in the diagnosis and the treatment of these rare benign bronchopulmonary tumours. CLINICAL CASE: Recent pneumopathy in the right superior lobe bronchial area in a 73 year old female, symptomatic with cought. Biopsy exeresis gave the histological diagnosis. DISCUSSION AND CONCLUSION: The clinical symptoms due to bronchial obstruction associated with the conventional radiological and computed tomography signs orientate the diagnosis definitively obtained through the anatomopathological study. This histological study is carried out from biopsies performed either during a bronchial fibroscopy or during a tracheobronchoscopy. By confronting the benign macroscopic aspect and the data from the extemporaneous anatomopathological study of the lesion, endoscopy provides an early-stage diagnosis and makes it possible to perform a concomitant and conservative treatment before the stage of irreversible broncho-pulmonary complications. It also avoids the risks and sequelae of classical surgery by thoracotomy. PMID- 16366381 TI - [Pseudoaneurysm of the internal carotid artery revelated by epistaxis: report of two cases]. AB - OBJECTIVES: Pseudoaneurysms of the carotid artery are a rare but serious cause of epistaxis. They can be rapidly life-threatening. Usually resulting of a surgery or a traumatism, pseudoaneurysms can express themselves by recurrent epistaxis. Management of these epistaxis is discussed. METHODS: The authors present two cases and effect a review of the literature. RESULTS: The first patient presented with a pseudoaneurysm few days after a transsphenoidal surgery. The second patient presented with a pseudoaneurysm fourteen years after a radio-surgical treatment of an ethmoidal and frontal epidermoid carcinoma. Diagnosis was suspected during nasal endoscopy and confirmed in both cases with angio-CT and arteriography. The first patient underwent a selective embolization. The second patient had to have a carotid occlusion due to a sudden severe haemorrhage, resulting from a pseudoaneurysm rupture following the embolization attempt. CONCLUSION: Review of the literature confirms the indication of endovascular intervention. The optimal management is carotid occlusion, provided the patient can tolerate carotid test occlusion and the circle of Willis is satisfactory. In some cases, a preservative treatment may be possible, which could be either pseudoaneurysm embolization or the placement of a covered stent. PMID- 16366382 TI - What is the effect of dizziness on the quality of life for patients with Meniere's disease? AB - OBJECTIVE: To evaluate the impact of dizziness on quality of life (QOL) in Meniere's disease (MD) patients. METHODS OF STUDY: Fifty definite MD patients were submitted to the Dizziness Handicap Inventory (DHI), in and out of crisis. The DHI scores were correlated to age, sex, race, disease duration, unilateral/bilateral labyrinth involvement, duration/frequency of vertigo attacks, vestibular and auditory function, aural fullness, tinnitus, imbalance and functional disabilities. Statistical analysis was performed by ANOVA. RESULTS: During crisis, all DHI aspects scores were statistically higher (p < 0.001) than out of crisis. Out of crisis, a statistically significant correlation was found between physical, functional and total scores and bilateral involvement. CONCLUSIONS: QOL impairment is worse during a Meniere's crisis. Out of crisis, QOL impairment is greater in patients with bilateral involvement. PMID- 16366383 TI - A report of two cases of unilateral vestibulopathie after systemic ototoxic treatment. AB - PURPOSE OF THE STUDY: To present the cases of two patients that developed unilateral vestibular hypofunction after systemic treatment with ototoxic agents, although they manifested symptoms more typical of bilateral vestibular hypofunction. MATERIAL AND METHODS: The patients were seen after having recovered from their initial illnesses, and both of them denied suffering any spells of vertigo, loss of hearing during the treatment or tinnitus. RESULTS: In both patients, oscillopsia and vestibular ataxia were of varying intensity. Bedside vestibular examination, caloric and rotatory chair testing, and vestibular evoked myogenic potentials were congruent with a complete unilateral loss of vestibular function. Audiometry was normal in one case whereas in the other, there was a moderate bilateral sensorineural loss of hearing that was present before treatment and that did not change during the course of the treatment. CONCLUSION: The existence of unilateral vestibular loss was an unsuspected finding but after careful bedside examination, it was confirmed through extensive vestibular testing. However, this infrequent finding responded very well to vestibular rehabilitation. Different mechanisms are proposed to explain this phenomenon, although there is still no clear evidence of which may account for the responses observed. PMID- 16366385 TI - Influence of the relapse and remission periods on hearing status in children with minimal change nephrotic syndrome. AB - OBJECTIVE: Minimal change nephrotic syndrome (MCNS) is characterized by the onset of NS (Nephrotic Syndrome) without systemic disease, hypocomplementemia, or other serious signs of renal disease. Hearing status is not very well known in MCNS. Our objective was to address this question and to find out remission and relapse periods of the syndrome would affect the hearing of the patients. METHODS AND PATIENTS: Otologic status of 26 children with clinical MCNS was investigated in relapse and remission periods using audiometry and tympanometry. The pure tones that were obtained at the frequencies 250, 500, 1000, 2000, 4000 and 6000 Hz were noted. Pure tone averages (PTAs) were calculated at 500, 1000, 2000 and 4000 Hz frequencies. RESULTS: In both remission and relapse periods, PTA of the patients did not change and was 13 dB. The frequency specific pure tone results were not significantly different between the right and left ears of the patients as well as between the remission and relapse periods (p > 0.05). In the relapsing and remission periods, type A tympanogram was encountered in 86.4% and 92.3% of the ears, respectively. Type B tympanogram was encountered in 11.5% and 3.8% of the ears in the relapsing and remission periods, respectively. Type C tympanogram was encountered in 3.8% of the ears both in the relapsing and remission periods. Differences between the tympanometry results were not significant (p > 0.05). CONCLUSION: MCNS in childhood is not associated with an alteration in the hearing status, both in remission and relapse periods of the disease. PMID- 16366386 TI - [Comparative results of type II ossiculoplasty: incus transposition versus titanium PORP (Kurz)]. AB - OBJECTIVE: The goal of this study was to compare the anatomical and functional results of two groups of patients having an ossiculoplasty type II, either with the incus, or with a PORP Bell Vario prosthesis (Kurz). MATERIALS AND METHODS: Retrospective study concerning 98 patients having had an ossiculoplasty type II with the incus (Incus group) and 50 patients having received a PORP titanium prosthesis Bell Vario from Kurz (Titanium group). RESULTS: The rate of extrusion was small (1% Incus group and 4% Titanium group). Average preoperative air bone gap of patients who received the Titanium prosthesis was 21.2 dB and 42.8 dB for the incus group. The average postoperative air bone gap was 16.9 dB Titanium group and 25.5 dB Incus group (p < 0.05). The gain was 18.7dB in the Incus group and 4.3 dB in Titanium group (p < 0.05). The difference between pre and post air conduction thresholds in both groups was statistically significant (p < 0.05). The post-op airbone gap was less on 20 dB in 62% of cases (Incus group) and 61% (Titanium group). There was no significant loss of the bone conduction thresholds in each group. CONCLUSION: Audiometric results obtained among patients with an incus transposition are better than those obtained with a PORP Titanium prosthesis. These results are to be appreciated by taking account of the greatest proportion of cholesteatoma and of reoperation in the Titanium group. The PORP titanium Bell Vario prosthesis (Kurz) is a material well tolerated by the middle ear as shown by the low rate extrusion. Its qualities (easy and precise measurement, easy positioning) make a material of choice of it when the patient's incus is not usable. PMID- 16366384 TI - [The outcome of treatments for carcinoma of the external auditory canal]. AB - OBJECTIVE: A retrospective analysis of management and survival of patients treated for temporal bone carcinoma. PATIENTS AND METHODS: Thirty patients underwent treatment for carcinoma of the temporal bone. Twenty-five squamous cell carcinomas, 1 melanoma, 2 basocellular carcinomas and 2 adenoid cystic carcinomas were treated. Thirteen patients were treated before for the same disease. RESULTS: Staging revealed 12 T1 and T2, 6 T3 and 12 T4 tumours. The mean follow up was 5 years (2-276 months). The Kaplan Meier survival curves showed survival rates at 2 years of 82%, 67% and 32%, and at 5 years of 82%, 67% and 17%, respectively for the stages T1 or T2, T3 and T4. At the end of follow up at 9 years the survival rates were 66%, 66% and 17% for the stages T1 or T2, T3 and T4 respectively. Overall stages a complete remission was found in 65% and 23%, and deceased was 35% and 77%, respectively for the primary treatment group and the salvage surgery group. CONCLUSION: Long-term prognosis of the carcinoma of the external auditory canal mainly depends on the stage and primary treatment. Surgery (lateral temporal bone or subtotal temporal bone resection, both in combination with a neck dissection and a parotidectomy) and adjuvant radiotherapy is the treatment of choice for part of stage T1 and all T2 and T3 tumours. The improved survival (65%) of patients treated de novo compared with those treated with salvage surgery (23%) suggests that early referral and aggressive primary surgical treatment with postoperative radiotherapy offer the greatest chance of cure. PMID- 16366387 TI - [Reinforcing tympanoplasty with cartilage mosaic (differences from the palisade technique)]. AB - OBJECTIVES: To demonstrate the interest of cartilage mosaic tympanoplasty in the reconstruction of the tympanic membrane. MATERIALS AND METHODS: Retrospective series of 103 patients; the authors analyzed their results after cartilage mosaic tympanoplasty, with a mean follow up of 3.5 years (3 to 7 years). They explain the surgical technique in detail. The average gain was calculated on the difference of the pre-and post-operative thresholds in air conduction on the four frequencies 500 Hz, 1000 Hz, 2000 Hz, 4000 Hz. RESULTS: The authors detail and analyze their anatomical and audiologic results. The total rate of closing of the tympanic membrane is 93.2%, accounting for 96 perforations closed among 103. A tympanic retraction sitting apart from the reinforced zone was noted in 0.97%, representative only one case of 103. Then the total rate of success is 92.23%, representing 8 failures of 103. The average pre- and post-operative air bone gap were 26.5 dB and 14.6 dB. The average gain was 12.5 dB (extremes of 0 dB to 40 dB). CONCLUSION: The authors show the great reliability of cartilage mosaic tympanoplasty to reconstruct the tympanic membrane. They widen the indications with all types of tympanoplasties. Especially with the recurrent perforations, and the perforations evolving in an inflammatory context and/or dysfunction of the Eustation tube. Their results show a major interest to use this technique in the anterior and inferior perforations, and whatever the ossicular chain status. PMID- 16366388 TI - [New technique of myringoplasty with cartilage in suspension--preliminary results]. AB - The authors describe a surgical technique of simple myringoplasty with a cartilage graft which can be done as a day surgery. The main point is the suspension of this graft by a suture (Vicryl 6-0) to the skin of the external auditory meatus. The preliminary results on twenty seven patients are successful with a rate of complete closure which is 92.6%. This technique must be reserved for dry, limited, nonmarginal perforations and without ossicular problem or connection with the handle of the malleus. PMID- 16366389 TI - [Focus on the BPPV: Semont maneuver for the horizontal canal and anterior canal- differential diagnosis]. PMID- 16366390 TI - Activation of tachykinin system in the pedunculopontine tegmental nucleus suppresses hippocampal theta rhythm in urethane-anesthetized rats. AB - The pedunculopontine tegmental nucleus (PPN) is one of the reticular generators of the hippocampal theta rhythm. The PPN neuronal circuitry related to theta generation involves its cholinergic, GABA-ergic and glutamatergic components. Here we provide data indicating that the PPN tachykinin system may also be a part of this circuitry. In the experimental model of the tail-pinch elicited hippocampal theta in urethane-anesthetized rats (implanted with bilateral recording electrodes in the stratum moleculare of the upper blade of the dentate gyrus and with injection cannula unilaterally inserted into the PPN) it was found that intra-PPN microinjection of Substance P (SP) and [d-Pro2, d-Phe7, d-Trp9] Substance P (DPDPDT) caused suppression of the theta and enhancement of the delta activity in the hippocampal EEG. Accordingly, there was approximately a 50% (SP) 70% (DPDPDT) decline of the peak power in the theta frequency range and a decrease by 0.4 Hz in the corresponding peak frequency (DPDPDT only) in both hippocampi. The circuitry through which SP exerts its effect in the PPN can be only hypothetical at present. We suggest SP-evoked activation (either direct or indirect through the glutamatergic inputs) of the GABA interneurons which may tonically inhibit PPN outputs to the other theta-relevant structures. PMID- 16366391 TI - Effects of maternal thiamine deficiencies on the pyramidal and granule cells of the hippocampus of rat pups. AB - To understand the implication of thiamine deficiency in the neuronal atrophy and cell death we undertook to induce thiamine (B1 vitamine) deficiency during three essential periods of the ontogenesis of rat central nervous system (CNS). Female rats were fed with a thiamine deprived diet during the gestation and lactation, and the fetuses and pups were alternately exposed to prenatal, perinatal or postnatal thiamine deficiencies. On the 45th postnatal day, histological studies were done on the brains of the pups and the structure of the hippocampus was analyzed. The effects of each treatment were assessed by measuring the size and the density of cell nuclei throughout the dentate gyrus and fields CA4, CA3 and CA1 of the hippocampal formation. The hippocampus showed a regional vulnerability in the pups exposed to maternal thiamine deficiencies. It appears that the thiamine deficiency decreased nuclear density (27.20%) more severely than nuclear size (10.56%) in the fetal hippocampus. Consequently, the major part of the teratogenic effects of thiamine deficiency was cellular death, rather than cellular atrophy. PMID- 16366392 TI - Maximizing the entropy of histogram bar heights to explore neural activity: a simulation study on auditory and tactile fibers. AB - Neurophysiologists often use histograms to explore patterns of activity in neural spike trains. The bin size selected to construct a histogram is crucial: too large bin widths result in coarse histograms, too small bin widths expand unimportant detail. Peri-stimulus time (PST) histograms of simulated nerve fibers were studied in the current article. This class of histograms gives information about neural activity in the temporal domain and is a density estimate for the spike rate. Scott's rule based on modem statistical theory suggests that the optimal bin size is inversely proportional to the cube root of sample size. However, this estimate requires a priori knowledge about the density function. Moreover, there are no good algorithms for adaptive-mesh histograms, which have variable bin sizes to minimize estimation errors. Therefore, an unconventional technique is proposed here to help experimenters in practice. This novel method maximizes the entropy of histogram-bar heights to find the unique bin size, which generates the highest disorder in a histogram (i.e., the most complex histogram), and is useful as a starting point for neural data mining. Although the proposed method is ad hoc from a density-estimation point of view, it is simple, efficient and more helpful in the experimental setting where no prior statistical information on neural activity is available. The results of simulations based on the entropy method are also discussed in relation to Ellaway's cumulative-sum technique, which can detect subtle changes in neural activity in certain conditions. PMID- 16366393 TI - On the role of mask structure in subliminal priming. AB - Choice reaction times to visual stimuli may be influenced by preceding subliminal stimuli (primes). Some authors reported a straight priming effect i.e., responses were faster when primes and targets called for the same response than when they called for different responses. Other authors found a reversed pattern of results. Our results suggest that the sign of the priming effect depends on mask structure. Inverse priming was obtained only for masks containing the searched for feature even though informational content of the masks was neutral. With masks of irrelevant structure, straight priming effects were found. Thus, masks are not passive stimuli whose roles are limited to rendering the prime invisible. Processing of the mask may interact with prime and target processing. Implications of the results are discussed for two hypotheses trying to account for straight and inverse priming (the self-inhibition hypothesis and object updating hypothesis). PMID- 16366394 TI - The need of neuroinformatic approach in functional neurophysiology. AB - Despite the enormous growth in the number of facts and regularities observed in neuroscience, the current state of the field does not allow their integration in coherent theoretical frameworks. Neuroinformatics is a new research field devoted to development of neuroscience data bases together with computational models and analytical tools for sharing, integration and analysis of experimental data and the advancement of theories of nervous system function. From 1996 the OECD countries promote fostering neuroinformatics by eliminating the barriers that prevent cooperation and by providing incentives to potential participants. This activity was formulated in the Report on Neuroinformatics from The Global Science Forum Neuroinformatics Working Group of the OECD (2002) as well as in several printed publications by the Neuroinformatics group (Amari et al. 2002, Eckersley et al. 2003). This article summarizes the ideas presented in the Report and stresses the importance of analytical and modeling approach to functional neurophysiology. PMID- 16366395 TI - Local classifiers for evoked potentials recorded from behaving rats. AB - Dynamic states of the brain determine the way information is processed in local neural networks. We have applied classical conditioning paradigm in order to study whether habituated and aroused states can be differentiated in single barrel column of rat's somatosensory cortex by means of analysis of field potentials evoked by stimulation of a single vibrissa. A new method using local classifiers is presented which allows for reliable and meaningful classification of single evoked potentials which might be consequently attributed to different functional states of the cortical column. PMID- 16366396 TI - Time-frequency-space localization of epileptic EEG oscillations. AB - This paper presents a hybrid method for localization of oscillatory EEG activity. It consists of two steps: multichannel matching pursuit with complex Gabor dictionary, and LORETA inverse solution. Proposed algorithm was successfully applied to the localization of epileptogenic EEG in a single patient. PMID- 16366397 TI - Analysis of multichannel biomedical data. AB - Nowadays, there is a common practice in biomedical research to perform multiple time series recordings. In the first part of this paper, basic information about analysis of such multichannel biomedical data is given. A short overview of important differences between single-channel, two-channel and multichannel data sets is presented and various coherence functions are reported. Causal relations between channels are investigated by means of the Directed Transfer Function (DTF) and its dynamic version, the Short-Time Directed Transfer Function (SDTF). The introduced formalism was used to analyze 12-channel human electrocorticogram (ECoG) records. Preliminary results of a study of causal dependence in beta and gamma frequency bands in two patients performing a motor task are reported. Specific characteristics in activity propagation consistent for both subjects for different rhythms were found. PMID- 16366398 TI - Novelty based feedback regulation in artificial neural networks. AB - In this paper we present a theoretical framework for novelty based feedback regulation in artificial neural networks. Novelty is assessed on the basis of monitoring the coherence of network dynamics. The result of novelty detection is dynamically coupled to parameters that control the dynamics of the recognition process. The paper presents a new measure of novelty detection--the strength of the local field--and presents new simulation results concerning novelty detection. It also integrates previously published models and simulation results into a general dynamical model of feedback regulation. PMID- 16366399 TI - [Preoperatory anxiety in children: psychological evaluation of premedication and parental presence efficacy]. AB - OBJECTIVES: The induction of anaesthesia for surgery is a stressful event for child. To treat pre-operative anxiety in children pharmalogical methods (premedication) and behavioural methods (the presence of parents during the induction of anesthesia) are used in combination with each other. The purpose of this investigation was to study the effect of two interventions to alleviate preoperative anxiety of 39 children who are undergoing minor surgery. We studied the influence of some psichological and social characteristics of child and parents on operative period. METHODS: We studied 39 italian speaking children aged 2-14 years ASA 1. We used STAI, a semi structured interview, with questions in the social-demographical area, a valuation scale of the behaviour of the child completed by the anesthesist, a questionaire after surgery for the parent. The descriptive analysis has been performed taking into account the nature of variables (frequency distributions for nominal and ordinal variables; means and standard deviations for cardinal variables). RESULTS: The results show that the age of the child is strongly conditional to the level of stress: negative reactions are more usual in children of pre-school age. The sedative premedication is not an important factor in relieving anxiety, and this result contrasts with published studies, stating the premedication is more effective than presence of a parent. We obtained interesting results by relating the level of stress in the child with other variables: which parent, the level of stress according to the STAI, level of education, type of residence of parents. CONCLUSIONS: Mother's presence during induction resulted of paramount importance. This study shows that the premedication alone is not effective. Some social demographical factors of the parents have influence. PMID- 16366400 TI - [Natural history of prenatally diagnosed hydronephrosis: possible recurrence of dilatation after spontaneous reduction, in cases with pelvic diameter larger than 20 mms, recommends long term follow-up]. AB - BACKGROUND: Long term follow-up of a prenatally diagnosed hydronephrosis usually extends no longer than the first two years of life. During this period spontaneous reduction occurs in most of the dilatations, not sustained by obstruction or reflux. Late recurrence of hydronephrosis is considered to be unusual. The aim of the present work has been to verify the risk of recurrent hydronephrosis long time after reduction and to identify factors associated to recurrence. MATERIALS AND METHODS: In a seven years period (1992-99) 276 patients with hydronephrosis unrelated to reflux, duplex kidney, megaureter or vesical obstruction have been observed. Among them, 231 were referred after prenatal diagnosis. Hydronephrosis of grade III or more was recorded in 73/231 with a pelvic diameter > or = 15 mms at ultrasonography (US) and a normal counter lateral kidney. Surgical treatment was elected in 39 cases on the basis of a separate function < 40%, deterioration during follow-up, or occurrence of clinical symptoms. Thirthy four cases were treated conservatively and four were lost at follow. Among the remaining 30 cases, 14 had a pelvic diameter lager than 20 mms. They were followed for a mean of 16.6 months (range 7-26) and spontaneous significant reduction of pelvic dilatation was recorded in all of them. RESULTS: Patients were recalled after a mean of 32.1 months from the last US. In 3 cases among 14 with a pelvic diameter larger than 20 mms a recurrent severe hydronephrosis was found respectively at 39, 56, and 68 months. In two of them, isotopic scans documented a reduced separate function. The third case reported recurrent symtpoms. A pyeloplasty was performed in all cases. CONCLUSIONS: Spontaneous reduction in most of the prenatally detected cases of neonatal hydronephrosis within the first two months of life make follow-up beyond this term controversial, whenever complete disappearance of pelvic dilatation has been documented. The Authors report three cases with severe hydronephrosis (pelvic diameter > 20 mms) among a group of patients treated conservatively. Long time after spontaneous reduction, recurrent hydronephrosis was documented in all by US, associated to deterioration of separate function and symptoms. Long term follow-up is recommendable in severe cases of prenatally detected hydronephrosis cases, even after reduction, to warrant from recurrence and renal damage. PMID- 16366401 TI - [Epidemiology of tonsillectomy and/or adenoidectomy in Italy]. AB - OBJECTIVES AND METHODS: Aim of the study was to describe frequency, causes, regional variations, setting and risk of mortality of tonsillectomy and adenoidectomy in Italy. The study is based on hospital discharge data for the years 1998-2000 provided by the Ministry of Health. RESULTS: During the year 2000, 61.280 tonsillectomy with or without adenoidectomy and 32.655 adenoidectomy alone were performed in Italy. ICD-9-CM codes reported on discharge abstracts indicated that the most frequent causes of tonsillectomy were chronic tonsillitis (45%) and hypertrophy of tonsils and adenoids (43%). The total tonsillectomy rate was 10.6 x 10.000 (CI 10.5-10.7) in 2000, and it was stable throughout the study period. We observed a wide geographical variability of regional tonsillectomy rates, standardised by age and sex. They ranged from 3.5 x 10.000 (CI 3.1-4.0) in Basilicata to 19.0 (CI 18.6-19.5) in Piemonte. We found an inverse correlation (r = -0.50) between regional tonsillectomy rates for tonsillitis and minimum temperatures recorded in capitals of the regions. Most operations were performed in the acute setting with a hospital stay longer than one day. The mortality risk associated to surgery was estimated to be at least of one case over 95.000 operations. CONCLUSIONS: The observed variability of regional tonsillectomy rates, only partly explained by weather differences, may be ascribed to disagreement among physicians. Inappropriate variations and setting were the two main reasons conducive to the development of the guidelines "Clinical and organisational appropriateness of tonsillectomy and/or adenoidectomy in Italy", in the frame of LINCO project and of the Italian National Program for Guidelines. PMID- 16366402 TI - The lazy bladder syndrome: a possible urodynamic evolution in patients with idiopatic detrusor and pelvic floor overactivity. AB - OBJECTIVE: To correlate the urge syndrome due to bladder overactivity and the lazy bladder syndrome, demonstrating that, at least in some cases, the lazy bladder may be the final stage of the evolution of an overactive bladder when associated with overactivity of the pelvic floor during micturition. PATIENTS AND METHODS: From January 1998 to December 1999, 38 children, 30 females and 8 males, 5 to 16 yrs. old (median 7.4 yrs), presenting with urge symptoms and never treated before, have been evaluated with repeated urodynamic investigations. At presentation all the patients underwent complete baseline urodynamics including evaluation of free flow, EMG, cystometrogram, subtracted detrusor pressure and flow/pressure studies. Flow/EMG was repeated every four months during the treatment period and full urodynamic investigation every year. RESULTS: The first urodynamic study showed that 17 patients were affected by pure detrusor overactivity with good detrusor-sphincter co-ordination, while 21 presented both detrusor and pelvic floor muscles overactivity. All the patients have been treated with oxibutinin (0.3-0.5 mg/Kg. in 3 divided doses) and a timed voiding program with the help of a frequency-volume chart. At the end of the study 16 out of the 17 children with pure overactive bladder were cured (94%), while only 12 of the 21 patients with both bladder and sphincter overactivity were clinically and urodinamically normal Nine girls showed a progressive shift towards the development of a lazy bladder syndrome (capacious, hypocontractile bladder with large post-voiding residual and a non relaxing sphincter during micturition). Clinically this shift was signalled by recurrent urinary tract infections. CONCLUSIONS: The transition from an overactive bladder to a hypocontractile one has been reported previously, but it has not been extensively investigated in children and its causes are largely hypothetical. In our cases this phenomenon occurred only in girls that, initially showed both detrusor urethral sphincter overactivity, therefore we can reasonably conceive that, at least in some cases, the lazy bladder syndrome may be due to prolonged and repeated high pressure bladder contractions opposed by a non relaxing sphincter that may lead to a progressive loss of the contractile properties of detrusor muscle fibres. The rather rapid shift observed in our patients might have been favoured by the use of anticholinergic medications. PMID- 16366403 TI - [Severe combined immunodeficiencies: a case of adenosine-deaminase deficit]. AB - Severe combined immunodeficiencies (SCID) are a group of rare genetic disorders characterized by profoundly defective T lymphocyte. We described in a two months old male a case of SCID with ADA deficiency. With this new case report we summarize recent developments in immunodeficiencies therapy, aiming to induce to bear in mind this disorder, despite its rarity, in differential diagnosis of infections, particularly respiratory or gastrointestinal infections. PMID- 16366404 TI - Partial idiopathic epilepsy: recovery after allergen-free diet. AB - The hypothesis that certain foods or allergens might bring about convulsions has been suggested repeatedly in the literature over the last century. Some clinical studies have highlighted an unusually high prevalence of allergic disorders in patients with epilepsy. This paper reports the consistent disappearance of partial idiopathic epilepsy symptoms in a nine-year-old patient as a result of diet free of cow's milk protein. This case appears to confirm the possible role of food allergy in certain types of epilepsy in patients of pediatric age. PMID- 16366405 TI - [Multiple sclerosis and acute disseminated encephalomyelitis in children: a difficult diagnostic differentiation. Report of a young boy with early onset of the disease]. AB - We report the case of an eight year old boy who developed suddenly acute left sided hemiparesis syndrome. Brain magnetic resonance imaging (MRI) showed multiple white matter lesions. Therefore we considered in the differential diagnosis multiple sclerosis (MS) and acute disseminated encephalomyelitis (ADEM). The patient received intravenous immunoglobulin (IVIG), 1 g/kg/d over 2 days with complete regression of clinical symptoms. No relapses occurred within six months, although brain magnetic resonance imaging studies found new white matter lesions, suggesting multiple sclerosis with very early onset. PMID- 16366406 TI - [Rhino-pharyngeal and middle ear diseases in children]. PMID- 16366407 TI - [Acute disseminated encephalomyelitis and multiple sclerosis]. PMID- 16366408 TI - [Prophylaxis of group B streptococcal infections in the birth centers of Emilia Romagna]. AB - OBJECTIVES: In the last decade several guidelines for prevention of neonatal group B streptococcal invasive disease have been published, mainly based on administration of intrapartum antibiotics. The spread of such recommendations yielded a reduction of the early-onset disease. The aim of the study was to investigate the practices for prophylaxis of neonatal infection in our region during the 2000 and to standardize them according to the new available evidence. METHODS: We conducted a multicenter study in Emilia Romagna, sending by mail a detailed questionnaire to the 28 birth centers of our region. RESULTS: Fifteen centers answered to the questionnaire. The practices were often differing from the recommendations of scientific societies. The most sensitive methods to identify colonized women were not widely used. Colonized infants were frequently treated with antibiotics and discharged later from the nursery. The incidence of neonatal invasive diseases was low, but most centers did not regularly collect a blood culture before antibiotic treatment is started. CONCLUSIONS: Repeated meetings among centers promoted the diffusion of information, the implementation of a shared protocol and the spread of the practices. The results of such meetings will be evalued in the next months. PMID- 16366409 TI - [Efficacy of early cerebral MR in the detection of brain lesions in high risk preterm infants compared with conventional US]. AB - To identify the efficacy of early cerebral MR, performed in the first month of birth, in the detection of brain lesions in high risk preterm infants, compared with conventional US, we recruited into the study a group of 30 preterm infants born at or below a gestational age of 30 weeks, who had a pathologic scan. The findings on US were compared with those of the early MR scan, performed in the same days, the results of which were considered as the final diagnosis. The value of cranial US as a predictor of MR signal intensity was assessed by calculating sensitivity, specificity, positive and negative predictive values. Agreement between two investigations was evaluated by calculating the K coefficient. US showed 33 haemorrhagic lesions in 25 preterms; MR showed 27 haemorrhagic lesions in 22 infants: in 16 cases MR gave the same results of US. Cranial US was reliable in detecting lesions such as GLH and IVH, but less sensitive in the definition of their size and distribution. Sensitivity of US for haemorrhagic lesions was 96.3%, PPV 78.8%, K coefficient 0.55 (p < 0.001). About the White Matter, cranial US demonstrated 20 lesions in 20 preterms; MR showed 16 lesions in 16 infants: in 3 cases MR was agree to US. US showed high reliability in the detection of cystic lesions, but significant limitations in the demonstration of non-cystic injury. We founded that normal WM echogenicity on US is not a good predictor of normal WM signal intensity on MR (30%). Sensitivity of US for WM lesions was 81.3%, PPV 65%, K coefficient 0.23 (p = 0.04). Finally US showed 4 lesions in other brain locations, MR confirmed 3 of them and discovered other 10. Sensitivity of US for these lesions was 23.1%, PPV 75%, K coefficient 0.21 (p = 0.11). We founded that cranial US is a good method for detecting GLH, IVH, HPI and severe WM lesions (cystic PVL), but it can miss non-cystic PVL, punctate haemorrhages, WMD and lesions in other brain locations, that, on the other hand, MR detects clearly. PMID- 16366410 TI - [Congenital megacolon in neonates and infants: impact of early, one-stage repair on morbidity and surgical complications]. AB - Hirschsprung's disease (HD) is a major cause of congenital bowel obstruction in children. Classical management includes stoma creation, a pull-through procedure, and restoration of bowel continuity. Colostomy may be also pulled through directly at second operation. Since the first report of a single stage (SS) primary pull-through without colostomy for HD, in 1980, several patients have been treated according to this procedure, even in neonatal period. All of them had a favourable outcome with better compliance of families and less hospital stay. The Authors reviewed the clinical records of 22 patients selected among 36 treated for HD between 1992 and 2000; in all, disease extended to rectum-sigma descendent; all cases of "total aganglionosis" or "ultra short" cases were excluded. Fifteen patients underwent SS repair: 11 neonates were operated at mean age of 69 days (range 14-172). In four infants diagnosis was made later (mean age 15 +/- 9 months) and SS repair was performed 8 days after (range 4-14). Seven patients underwent multiple stage (MS) repairs for clinical reasons: pull-through of the previous colostomy was performed at mean age of 171 days (range 47-330). A Duhamel procedure with an Endo-GIA linear stapler was performed in all No differences in gestational age or birth weight were found in both groups. Complications were observed only in the MS group (2 related to colostomy, 1 to postoperative adhesions). The Authors conclude that primary SS treatment of HD is safe and effective even in newborn patient, reduce familiar stress and hospital stay. The lower morbidity and complication rates could be explained by the limited bowel manipulation. Accurate histological-hystochemical diagnosis and meticulous preoperative bowel management should be part of the management. PMID- 16366411 TI - [Primary bladder diverticula in children: report of 15 cases]. AB - The Authors report their experience in primitive bladder diverticula treatment during the last six years. They identify criteria for a general classification of this clinical entity and limited to primitive forms they divide in 4 groups. This classification is related to clinical and topographycal criteria, aiming to delineate a new diagnostic and therapeutic pathway. PMID- 16366412 TI - [Transanal endorectal pull-through with or without laparoscopic assistance? Development of an experimental model]. AB - The transanal approach is a relatively new technique for surgery of Hirschsprung's disease. The purpose of this experimental study was to evaluate and compare the transanal endorectal pull-through when it takes place with laparoscopic assistance, according to Soave-Georgeson procedure and without laparoscopic assistance. We used two separated groups of animals. In the 1st group the transanal endorectal pull-through was performed by retrograde transanal mucosectomy with laparoscopic devascularization of the colon and in the 2nd group the transanal endorectal pull-through was performed without laparoscopic assistance. In the 1st group the transitional zone of the colon was determined laparoscopically while in the 2nd group during the perineal devascularization of the bowel. In both groups end to end coloanal anastomosis was performed in the same way. The postoperative course was uneventful, no severe long-term problems were observed. Four months after operation the experimental subjects of the 1st group had 3-4 discharges in 24 hours. While the experimental subjects of the 2nd group had 5-7 discharges in 24 hours. Manometric resting anal sphincters pressure in the 1st group averaged 77 +/- 16 mmHg compared with 74 +/- 15 mmHg in the 2nd group. The transanal colectomy technically can be performed with or without laparoscopic assistance. However the advantages of laparoscopic assistance are the following: 1) more exact determination of transitional zone; 2) easier separation of peritoneal reflection, 3) better control of final position of the colon; 4) minor retraction on the perineal muscles. PMID- 16366414 TI - [Imaging preterm babies' brain with MRI]. PMID- 16366413 TI - Annular pancreas combined with distal stenosis. A report of four cases and review of the literature. AB - BACKGROUND: Congenital duodenal obstruction (CDO) in combination with more distal duodenal obstructions is a rare anomaly occurring in 4% of neonates with duodenal atresia. The experience of two European Pediatric Centers in treatment of congenital double duodenal obstruction (CDDO) is reported and the pertinent literature is reviewed. MATERIALS AND METHODS: During the last 15 years a total of 86 neonates were operated upon for CDO at the department of pediatric surgery of "St. Sophia" Children's Hospital of Athens in Greece and the department of pediatric surgery of "Federico II" Children's University Hospital of Naples in Italy; four of them had a CDDO. These ones presented with nonbilious vomiting and the plain film of the abdomen showed the typical "double bubble". Our cases with CDDO presented annular pancreas causing complete obstruction of the second part of the duodenum and dilatation of the duodenum distal to this obstruction due to an additional congenital stenosis (two cases) or a membranous web (two cases). A diamond shaped duodeno-duodenal (DDD) anastomosis was carried out to relieve the proximal obstruction and a Heinecke-Mikulicz plasty was used to relieve the distal stenosis. RESULTS: All patients with double obstruction underwent successful surgery with no complications. The postoperative course was uneventfuL An upper gastrointestinal barium study at one month postoperatively showed no blind loop, megaduodenum, anastomotic stenosis or malfunction. CONCLUSIONS: a) The combination of duodenal atresia with annular pancreas and distal duodenal stenosis or web is extremely rare. b) Patency of the duodenum distal to the usual obstruction should always be checked in order to avoid misdiagnosis of this combination. PMID- 16366415 TI - [Special considerations for the prevention of perinatal group B streptococcal disease]. PMID- 16366416 TI - Prevalence of DRB1*01 and DRB1*04 alleles in a group of patients with rheumatoid arthritis living in Liguria. AB - A major component of genetic susceptibility to rheumatoid arthritis (RA) appears to be explained by inheritance of HLA-DRB1 alleles. Multiple HLA-DRB1 alleles (DRB1*0401, *0404, *0405, *0408, *0101, *102, *1001 and *1402) encoding a shared epitope at amino acid positions 70-74 are associated with susceptibility to RA. There is ethnic variation in the clinical expression of RA and in both the frequency and type of HLA-DRB1 alleles carrying the shared epitope. We evaluated the prevalence of the alleles of HLA-DRB1 locus encoding for SE in 42 outpatients with RA attending the Rheumatology Center of the University of Genoa, Bruzzone Institute, and living in Liguria. A control group was composed of Italian marrow donors. DNA genotyping was performed using a low-resolution polymerase chain reaction technique for characterization of the families of HLA-DRB1 alleles for each of the 42 patients studied. Subsequently, subjects with *01 and *04 haplotype were tested with high-resolution HLA-DRB 1 typing to characterize the *01 and *04 alleles. No statistically significant differences were found in the prevalence of RA-associated single alleles *01 and *04 in the study group or in the control group. In contrast, the sum of susceptibility *04 alleles studied by resolution typing was strongly related to RA in the study group in comparison with the control group. PMID- 16366418 TI - Attenuated insulin response and normal insulin sensitivity in lean patients with ankylosing spondylitis. AB - Chronic low-grade inflammation is associated with insulin resistance. The aim of this study was to determine insulin response to intravenous glucose load and insulin sensitivity in patients with ankylosing spondylitis (AS). Fourteen nonobese male patients with AS and 14 matched healthy controls underwent frequent sampling intravenous glucose tolerance test (FSIVGTT). Insulin secretion and insulin sensitivity were calculated using the computer-minimal and homeostasis model assessment 2 (HOMA2) models. Fasting glucose, insulin, cholesterol, high density lipoprotein and low-density lipoprotein cholesterol, triglyceride levels, HOMA2, glucose effectiveness, insulin sensitivity and insulin response to FSIVGTT did not differ between patients and controls. Tumor necrosis factor-alpha and interleukin (IL)-6 concentrations tended to be higher in AS patients than in controls. Second-phase beta-cell responsiveness was 37% lower (p = 0.05) in AS patients than in controls. A negative correlation was found between the percentage of beta-cell secretion and IL-6 in all subjects (r = -0.54, p = 0.006). We found normal insulin sensitivity but attenuated glucose utilization in the second phase of FSIVGTT in AS patients. Our results indicate that elevated IL 6 levels may play a pathophysiological role in attenuating beta-cell responsiveness, which may explain the association between elevated IL-6 levels and increased risk for type 2 diabetes. PMID- 16366417 TI - Methotrexate-induced acute lung injury in a patient with rheumatoid arthritis. AB - Methotrexate (MTX)-induced acute lung injury developed in a female patient with rheumatoid arthritis. She was successfully treated with high-dose glucocorticoid therapy. During her hospital stay, the serum concentration of surfactant protein (SP)-D, which was markedly elevated on admission, was finally normalized and the disease resolved. However, the serum concentration of Klebs von den Lungen (KL)-6 remained high. Although the mechanisms of lung injury by MTX have not been well defined, serial measurements of serum SPD might be useful for the clinical evaluation of drug-induced acute lung injury. PMID- 16366419 TI - Phase II study of a novel oral formation of 5-fluorouracil in combination with low-dose cisplatin as preoperative chemotherapy of oral squamous cell carcinoma. AB - TS-1 is a novel oral 5-fluorouracil containing tegaful (prodrug of 5-FU) and two biochemical modulators. These modulators feature effect-enhancing and adverse reaction-reducing activity. We investigated the histological response and toxicities of combination chemotherapy with TS- 1 and low-dose cisplatin and evaluated its usefulness as preoperative chemotherapy Forty-four newly diagnosed patients with stage Il-IV oral squamous cell carcinoma were enrolled in this study from February 2002 to April 2004. Patients were administered TS-1 80 mg/m2/day (days 1-14) and cisplatin 5 mg/m2/day (days 1-5 and 8-12) followed by radical surgery within 2 weeks. The histopathological effect of chemotherapy, which was a surrogate endpoint of this trial, was evaluated with surgical or biopsy specimens. The rate of histological antitumor effect was as follows: complete response (CR) 36.4%, partial response (PR) 25.0%, minor response (MR) 18.1% and no change (NC) 20.5%. The rate of histological response (CR + PR) was 61.4%. The CR rate of effective cases was 59.3%. The main toxicities occurred in bone marrow and the digestive tract. The incidence of severe toxicity such as grade 3 or 4 was 4.5% in anemia, 9% in leukocytopenia, 11.4% in neutropenia, 4.5% in thrombocytopenia and 2.3% in anorexia, diarrhea and urticaria. Most patients showed no toxicity or mild toxicities. TS- 1 with low-dose cisplatin has highly effective antitumor activity and mild toxicities. In particular, the CR rate was very high. It is suggested that this regimen is suitable for neoadjuvant chemotherapy. We expect that this chemotherapy will contribute to avoidance of surgery for small tumors (stages I and II) and will enable function-preserving surgery for advanced tumors. PMID- 16366420 TI - High bone-binding capacity of ibandronate in hemodialysis patients. AB - Bisphosphonates are a potential therapy for osteoclast-mediated bone disease, such as renal osteodystrophy. This study evaluated ibandronate bone-binding in patients with secondary hyperparathyroidism and renal osteodystrophy and examined whether there is a correlation with bone metabolism parameters. Sixteen patients with end-stage renal disease and secondary hyperparathyroidism receiving regular hemodialysis were recruited to this 12-week trial. Intravenous ibandronate 2 mg was administered for 5 min every 4 weeks directly after hemodialysis. Ibandronate levels were measured 15 min after infusion and at trough levels before the next hemodialysis. Serological markers of bone metabolism were also measured. After the first infusion, the peak ibandronate level was 154 +/- 75.1 ng/ml and the trough level was 2.7 +/- 1.7 ng/ml. At week 12, peak and trough ibandronate levels were 164.8 +/- 89.9 ng/ml and 3.2 +/- 2.6 ng/ml, respectively. Ibandronate bone uptake was 98.0% at first application and 98.4% at week 12. In patients with remaining diuresis, ibandronate urine excretion was < 0.001% of the administered dose. There was no correlation of ibandronate bone-binding with parameters of osteoclast activity or parathyroid hormone (PTH). The correlation with markers of osteoblast activity was significant but weak. Ibandronate had a bone-binding capacity of approximately 98% in hemodialysis patients. After repeated dosing ibandronate bone-uptake remained stable and was independent of osteoclast activity or PTH levels. Due to the high bone-binding of ibandronate in these patients, a 2 mg dose of intravenous ibandronate is equivalent to a 4-5 mg dose of ibandronate in patients with normal renal function. PMID- 16366421 TI - Efficacy of a novel calcium/potassium salt of (-)-hydroxycitric acid in weight control. AB - The weight-loss efficacy of a novel, water-soluble, calcium-potassium salt of (-) hydroxycitric acid (HCA-SX) was re-examined in 90 obese subjects (BMI: 30-50.8 kg/m2). We combined data from two previously reported randomized, double-blind, placebo-controlled clinical studies in order to achieve a better statistical evaluation based on a larger population. This re-examination of data also allowed us to reflect more intensely on various aspects of weight loss studies. Subjects were randomly divided into three groups: group A received a daily dose of HCA-SX 4, 667 mg (providing 2,800 mg HCA per day); group B was given a daily dose of a combination of HCA-SX 4,667 mg, niacin-bound chromium (NBC) 4 mg (providing 400 microg elemental chromium), and Gymnema sylvestre extract (GSE) 400 mg (providing 100 mg gymnemic acid); and group C received a placebo in three equally divided doses 30-60 min before each meal. All subjects were provided a 2,000 kcal diet/day and participated in a supervised walking program for 30 min/day, 5 days/week. Eighty-two subjects completed the study. At the end of 8 weeks, in group A, both body weight and BMI decreased by 5.4%, low-density lipoprotein and triglycerides levels were reduced by 12.9% and 6.9%, respectively, while high density lipoprotein levels increased by 8.9%, serum leptin levels decreased by 38%, serotonin levels increased by 44.5% and urinary excretion of fat metabolites increased by 32-109%. Group B demonstrated similar beneficial changes, but generally to a greater extent. No significant adverse effects were observed. The combined results confirm that HCA-SX and, to a greater degree, the combination of HCA-SX plus NBC and GSE reduce body weight and BMI, suppress appetite, improve blood lipid profiles, increase serum leptin and serotonin levels and increase fat oxidation more than placebo. We conclude that dosage levels, timing of administration, subject compliance and bioavailability of HCA-SX significantly affect results and that when taken as directed, HCA-SX is a highly effective adjunct to healthy weight control. PMID- 16366422 TI - Identification of human papillomavirus DNA in melanoma biopsy specimens of Greek population. AB - The aim of this retrospective study was to evaluate melanoma biopsy specimens from the Greek population living in the prefecture of Larissa for the presence of human papillomavirus (HPV) DNA and to determine the possible relationship between HPV and clinical outcome in these patients. Twenty-eight melanoma biopsy specimens, 20 from primary cutaneous melanoma and eight from melanoma metastasis were obtained from 28 patients. The biopsy samples were formalin-fixed and paraffin wax-embedded. The control group consisted of three junctional melanocytic nevi, histologically confirmed, and three punch biopsies from normal skin that were obtained from six healthy individuals. The presence and types of HPV DNA were assessed by the amplification of a fragment of the LI region by consensus primer polymerase chain reaction (PCR) combined with restriction fragment length polymorphism analysis (RFLPA). In each biopsy specimen that was evaluated, HPV 6, HPV 11, HPV 16 and HPV 18 positive controls from genital HPV lesions were included. Five of 28 (17.85%) biopsy melanoma specimens were positive for HPV DNA. Conversely, HPV was not detected in any of the biopsy specimens of the control group (0/6). HPV viral type 16 was found in two samples and HPV 6 DNA in three. Our results regarding the possible relationship between melanoma and HPV DNA were not statistically significant (p > 0.05). These findings suggest that ultraviolet sun exposure remains the main cause of melanoma in our region. The role of cutaneous HPV infection in the pathogenesis of melanoma remains elusive. PMID- 16366423 TI - Is there an association between clinical symptoms and intestinal parasitic infections? AB - Intestinal parasitic infections are major causes of childhood diseases in rural regions. The significant causes of this problem are the insufficient resources to improve the infrastructure in these regions and inadequate health services in both the diagnosis and treatment of diseases. The aim of this study was to assess the association between intestinal parasitic infections and clinical symptoms of infected children. Cellophane tape and stool samples of 100 children, aged between 7-10 years, from two villages of Manisa city, were examined for intestinal parasites and their symptoms were recorded. Although some symptoms, such as abdominal pain, anorexia and fatigue were found to be common in infected children, no significant relationship was found between symptoms and intestinal parasitic infections. PMID- 16366424 TI - Evaluation of hepatitis B surface antigen, anti-hepatitis C virus and anti-human immunodeficiency virus antibodies and syphilis seropositivity in blood donors: six years' seropositivity. AB - This retrospective study was carried out to evaluate the seropositivity of hepatitis B surface antigen (HbsAg), anti-hepatitis C virus (HCV), anti-human immunodeficiency virus (HIV) and syphilis in blood donors in Manisa Government Hospital. Data were evaluated in 10,189 blood donors between April 1, 1997, which is the time from which regular records began to be collected, and April 1, 2003. The blood samples of the blood center from April 1, 1997, to January 1, 1998, were examined via the card method, those between January 1, 1998, and January 1, 2002, were examined via micro enzyme-linked immunosorbent assay (ELISA) method and the rest were evaluated with macro ELISA methods. In blood donors, the positivity of HbsAg, anti-HCV anti-HIV and the rapid plasmin reagin test were 2.95%, 0.68%, 0.00% and 0.16%, respectively. PMID- 16366425 TI - 1-800-chronic disease management. PMID- 16366426 TI - No easy answers to "ten hard questions". PMID- 16366427 TI - No easy answers to "ten hard questions. PMID- 16366428 TI - No easy answers to "ten hard questions". PMID- 16366429 TI - No easy answers to "ten hard questions". PMID- 16366430 TI - No easy answers to "ten hard questions". PMID- 16366431 TI - No easy answers to "ten hard questions". PMID- 16366432 TI - No easy answers to "ten hard questions". PMID- 16366433 TI - No easy answers to "ten hard questions". PMID- 16366434 TI - No easy answers to "ten hard questions". PMID- 16366435 TI - No easy answers to "ten hard questions". PMID- 16366436 TI - No easy answers to "ten hard questions". PMID- 16366437 TI - Your clinical questions answered. PMID- 16366438 TI - A refresher on Medicare and concurrent care. PMID- 16366439 TI - Look beyond your practice's bottom line. PMID- 16366440 TI - Take your personal digital assistant to the next level. PMID- 16366441 TI - Choosing the right practice entity. PMID- 16366442 TI - Improving chronic illness care: lessons learned in a private practice. PMID- 16366444 TI - [Should disease management be feared?]. PMID- 16366443 TI - So how are you doing? PMID- 16366445 TI - [Should disease management be feared? (1): hospital care]. AB - The goals of disease management are: (1) an integrated health care delivery system; (2) knowledge-based care; (3) elaborate information systems; (4) continuous quality improvement. In-hospital disease management and, more specifically, critical pathways, establish standardized care plans, set goals and time actions to reach these goals. They can reduce variations in practice patterns and resource utilization without compromising quality of care. Such strategies participate to quality improvement programs in hospitals when they involve and empower all actors of a given process of care, are not imposed from outside, and use sound and rigorous development and evaluation methods. PMID- 16366446 TI - [Should disease management be feared? (2): outpatient care]. AB - Outpatient disease management is a multidisciplinary team intervention for managing complex processes of chronic diseases, in order to improve healthcare quality and decrease process variations. Interventions are based on: (1) evidence based guidelines; (2) educational programs; (3) close patient follow-up. This can be achieved by telephone follow-up, by outpatient clinic programs, or by homecare visits performed by case managers. For the management of patients with chronic heart failure, disease management programs have resulted in a 25% decrease in hospitalization and in reduced costs. In our Swiss health care system, however, a majority of patients are taken care of by private practitioners; thus, the involvement of these physicians in the development and in the realization of disease management programs will be key to their success. PMID- 16366447 TI - [Should B-type natriuretic peptide (BNP) be used in daily practice?]. AB - Each diagnostic test should be systematically evaluated before acceptation for use in daily practice. The principles for validating a diagnostic test are reviewed using B-type natriuretic peptide (BNP) as an example. BNP seems promising to diagnose symptomatic left-sided heart failure in the emergency department, especially when the diagnosis remains uncertain (intermediate clinical probability). However, before its widespread use, its efficacy on clinical outcome measures such as reduction of time to treatment or in-hospital length of stay should be more clearly demonstrated through pragmatic studies. PMID- 16366448 TI - [Educational strategies to modify physician behaviours and health care outcomes]. AB - The current evolution of medicine requires a sustained effort in continuing medical education to maintain an evidence-based practice. To support this process, several educational strategies are available, including guideline diffusion. Their critical analysis reveals that the dissemination of passive information by conferences or by unsolicited materials is generally ineffective to modify the behaviours in practice or patient outcomes. More effective interventions are practice- and community-based rather than didactic, include an evaluation of learning needs, and enhance active participation in multiple activities. Before investing time and money into an educational activity, one should consider the evidence of its efficacy and make an oriented choice, according to the goals aimed and the available resources. PMID- 16366449 TI - [Improving patient's medication knowledge at hospital discharge]. AB - The period following hospital discharge is a time of vulnerability regarding the continuity of care and carries the highest risk of patient errors and confusion with medications. Strategies to improve knowledge and understanding of medications at discharge are necessary and may result in safer use, better compliance, and reduced health care expenditures. In our service of general internal medicine, a structured patient-centered discharge interview, performed by the resident caring for the patient and reinforced by a simple and individualized treatment card, increased patients' knowledge of their discharge medications. PMID- 16366450 TI - [Nurse-physician collaboration: a determinant of the quality of care?]. AB - In hospital practice, the most elementary form of interdisciplinary care is the collaboration between nurses and physicians. Physicians rate the quality of collaboration with nurses higher than nurses themselves. Studies on the impact of interdisciplinary interventions on quality of care evaluate complex interventions and it is impossible to distinguish the effects of interdisciplinary care from other facets of the intervention. Two recent studies suggest that a better collaboration between physicians and nurses may improve patient outcomes, and reduce length of stay and hospital costs. What is meant by collaboration should be better defined to allow developing strategies for training health care professionals to collaborate efficiently and for evaluating their impact on health care quality. PMID- 16366451 TI - [Radical prostatectomy: open or "minimally invasive"? Our experience with open surgery and related comments]. AB - Radical prostatectomy remains the gold standard for treatment of localised prostate cancer. Standardisation of the open retro-pubic anatomic prostatectomy by P Walsh allows skilled but not expert surgeons to achieve a high standard of performance. Learning curve is short with this technic, with minor morbidity. Rates of incontinence are low and impotency is now rather uncommon in the younger patient while oncological control is optimal for histologicaly organ confined cancer. "Mini invasive technics", laparoscopy and robot-assisted laparoscopy, have a longer learning curve, including a higher rate of complications that are unusual with open surgery. Operating time remains longer, costs are superior to the open technic and oncological control is not yet clearly validated while rates of classical late complications are not lower. Consequently, most urologist still prefer the open approach PMID- 16366452 TI - [Is quality control efficacious?]. PMID- 16366453 TI - [Vaccination status in children: positive effects of introducing preschools visits by an attending physician]. PMID- 16366454 TI - [A billion dollars released by Geneva]. PMID- 16366455 TI - [Coca-Cola is causing allergy to obesity]. PMID- 16366456 TI - [Some news about developing an antimalarial vaccine]. PMID- 16366457 TI - [The appeal of generalists "who practice in cities where the revolt rumbles"]. PMID- 16366458 TI - [Calculating the impact of global warming on human health]. PMID- 16366459 TI - [Embezzlement?]. PMID- 16366460 TI - [Analysis on chemical and physical properties of Beijing super dust storm in spring of 2004]. AB - On March 27, 2004 a super dust storm occurred in Beijing. The concentration of the aerosol and the element concentration of the aerosol were analyzed. The velocity of wind and radiation of ground were also measured. TSP increased 300 - 400 percent. PM2.5 increased 200 - 300 percent, but the concentration of the fine particle decreased. The elements like Na, Mg, Al, Mn and Fe mainly distribute in coarse particles. The contaminated elements such as Zn and Pb mainly distribute in fine particles. The contaminated elements mainly come from local source. The total radiation reduced 37.8 percent. With the increased of the velocity of the wind, the concentration of the coarse particle increased notability, the concentration of the fine particle decreased. PMID- 16366461 TI - [Catalytic performance of Ce/Zr series catalysts on soot combustion]. AB - Catalytic performances of Ce/Zr series catalysts (Ce(x)Zr(1-x)O2) on soot combustion and the influence of feed gas were investigated by TG and TPO. The catalytic activity is high, and affects by the Ce/Zr ratio. The concentration of O2 affects the speed-limited step during the process of soot combustion. H2O showed no effect on the catalytic activity for soot combustion on Ce(0.5)Zr(0.5)O2. NO could promote soot combustion by presenting NO2, a more powerful oxidant than O2, and the ignition temperature of soot decreased 30 degrees C. Results of TG and TPO show that the beta species oxygen on the catalyst take part in the combustion process. PMID- 16366462 TI - [Preparation, characterization and three way catalytic performance for Pd/CZ/Al2O3 catalyst]. AB - Pd/CZ/Al2O3 catalyst was prepared by impregnating a noble metal solution to the support CZ/Al2O3 which was prefabricated by co-impregnation. The investigation results show that Pd/CZ/Al2O3 has a superior three-way catalytic performance, which is comparable to Pd/CZ for the fresh sample and a better one after thermal ageing. Based on the XRD, BET and TPR characterizations, the internal relationship between catalytic performance, composition and structure was discussed. The relatively high activity after thermal ageing is ascribed to the maintenance of the Strong Metal-Support Interaction (SMSI). PMID- 16366463 TI - [Emission and source characterization of monoaromatic hydrocarbons from coke production]. AB - Monoaromatic hydrocarbons (MAHs) from indigenous and industrial coking processes are studied in Shanxi province. They are sampled on the top of coke ovens and in the chimneys using stainless steel canister and determined by GC/MSD after preconcentration with liquid nitrogen. Benzene, toluene and xylene are the main components among MAHs emitted from coking processes. Benzene and the total MAHs concentrations were as high as 3421.0 microg/m3 and 4 865.9 microg/m3 in the air from indigenous coking, 548.7 microg/m3 and 1 054.8 microg/m3 in the oventop air from industrial coking, and 1 376.4 microg/m3 and 1 819.4 microg/m3 in stack gas from industrial coking, respectively. The MAHs concentrations vary greatly during the indigenous coking process, which in the prophase (from firing to 10 days) is obviously higher than in the anaphase (10 days to quenching the coke). In industrial coking the MAHs in the oventop air are highest when charging the coal and next when transferring the hot coke, but in stack gas they are highest when charging coal and lowest when transferring the coke. Benzene, toluene, ethylbenzene and xylene (BTEX) in industrial coking samples show good linearity, indicating that MAHs in industrial coking might come predominantly from coal pyrolysis; but BTEX distribute dispersedly in indigenous coking samples, indicating that its emission might be affected by many factors. In all samples BTEX ratios especially high B/E ratio, is unique among MAHs sources, and might be helpful to characterize pollution from coking. PMID- 16366464 TI - [Ecological distributions of airborne fungi in outdoor environments in Beijing, China]. AB - An investigation on fungal types, concentrations, and their dynamic variation in outdoor environments was carried out in three different functional areas around one year in Beijing. Results show that the fungal concentrations varied widely and the average was (1164.8 +/- 73.2) CFU x m(-3), ranging from 23.6CFU x m(-3) to 13 959.5 CFU x m(-3). The most common culturable airborne fungi in all seasons and all functional areas were Cladosporium , nonsporulating fungi, Alternaria, Penicillium and Aspergillus. The most dominant fungus was Cladosporium, which contributed to more than 1/3 of the total. The fungal levels in culture and education region (CER) and garden green region (GGR) were highest in the fall and summer, and lowest in the winter and spring, while the seasonal variation in main traffic line (MTL) was not significant. The fungal concentrations in CER and GGR were significantly higher than in MTL (p <0.05). No statistically significant difference exists between CER and GGR. PMID- 16366465 TI - [Assessment on ecological security spatial differences of west areas of Liaohe River based on GIS]. AB - Ecological security assessment and early warning research have spatiality; non linearity; randomicity, it is needed to deal with much spatial information. Spatial analysis and data management are advantages of GIS, it can define distribution trend and spatial relations of environmental factors, and show ecological security pattern graphically. The paper discusses the method of ecological security spatial differences of west areas of Liaohe River based on GIS and ecosystem non-health. First, studying on pressure-state-response (P-S-R) assessment indicators system, investigating in person and gathering information; Second, digitizing the river, applying fuzzy AHP to put weight, quantizing and calculating by fuzzy comparing; Last, establishing grid data-base; expounding spatial differences of ecological security by GIS Interpolate and Assembly. PMID- 16366466 TI - [Integrated fuzzy evaluation of water eutrophication based on GIS in the Taihu Lake]. AB - Supported by geographic information system and geostatistics, the application of fuzzy mathematics and analytic hierarchy process for water eutrophication evaluation was discussed. Taking Taihu Lake as an example, the research selected total phosphorus, total nitrogen, chlorophyll a, COD, BOD5, DO and transparence as evaluation index. After geostatistical analysis of the datum of monitoring site, the values of evaluation indices were estimated in the whole research area. Given that, the different dependence functions were developed for these indices and the function values were calculated. Furthermore, according to the principle of analytic hierarchy process, the weight of every index was calculated, then integrated evaluation value was obtained for the whole research area and the evaluation map for water eutrophication was drawn. The result shows that the level of nutrition is the highest in the north and north-west of lake, which is hypertrophic, that is meso-eutrophic in the middle and that is the lowest in the south-east of lake, which is mesotrophic. PMID- 16366467 TI - [Ecological protection and sustainable utilization of Erhai Lake, Yunnan]. AB - Economic development and increase of population pressure have caused a series of ecological environmental problems of Erhai Lake. These problems include: (1) Quickening of eutrophication process, (2) Decrease of water level and water resources, (3) Habitat deterioration of lakeside zone, and (4) Overfishing and slow depletion of aboriginal fish. Pollutant loading of Erhai Lake is as follows: COD(Cr) 3 008 t x a(-1), TP 137.31 t x a(-1), TN 1 426.35 t x a(-1). According to the mestrophic target of water quality, loading of nitrogen and phosphorus is far above environmental capacity of Erhai Lake. Erhai Lake is now in a pivotal and hypersensitive period of trophic states change, and the position is very critical. Therefore, some countermeasures to solve the problems are presented as follows: (1) Defining the dominant functions of Erhai Lake, (2) Paying attention to the adjustment of the industrial structure and distribution in the course of urbanization, (3) Setting up lakeside zone reserve, (4) Strengthening the control of tourism pollution, (5) Properly adjusting the water level of Erhai Lake, and (6) Some ecological engineering measures for water resources protection in the basin should be taken through collecting and treating of urban sewages, ecological rehabilitating of the main inflowing rivers, constructing of ecological agricultures and improving of rural environment, ecological restoring of aquatic ecosystem, and soil and water conservation. PMID- 16366468 TI - [Calculation of environmental flows in river reaches based on ecological objectives]. AB - Based on the identifying the ecological objectives, environmental flows in river reaches is calculated after the relation between parameters of objectives, and river discharges is determined. The ecological objectives are determined in a two step process: theobjective will be determined for the critical period of the year, then the temporal variation will be defined. Considering the compatibility between the different kinds of environmental flow requirements, the strictest objective is settled to be the ecological objectives for the critical period of the year. The temporal variation of the natural river discharge monthly is settled to be the temporal variation of ecological objectives. In the studies of the environmental flows in the river reaches downstream for Guanting reservoir in the Yongding River of Haihe River Basin, the requirements of velocities for spawning of the fish in April are regarded as the ecological objectives in the most critical period because this also is the period of highest water demand for irrigation. The relation between objectives and river discharge is identified using historical data at the river station. The results indicate that the minimum, medium and ideal lever of annual environmental flow requirements are 1.56 x 10(8) m3, 5.97 x 10(8) m3 and 11.02 x 10(8) m3, about 7.19%, 27.51% and 50.78 % of the natural river discharge respectively. The ratio of water requirements monthly should be 20 % in the flood period (Aug.) and be 20% in biological propagation period in spring (Apr. - Jun.). PMID- 16366469 TI - [Characteristic analysis on the anions and pH profiles of rainwater of Zhoushan Archipelago]. AB - Rainwater samples were collected at Zhoushan Archipelago in 2002 - 2003, 4 main anion concentrations (F-, Cl-, NO3-, SO4(2-)) and pH value of rainwater sample were determined. The concentration variation range and character were depicted. Principal component analysis (PCA) and factor analysis (FA) were used to acquire a further knowledge on the characteristics of anions and pH value. Distribution character of samples was clearly depicted in the new PC1-PC2 ordinate generated by orthogonal projection and the reasons that caused the outliers in samples were discussed. In order to get the latent variables which most distinguishably effect the sample distribution, data profiles were investigated by factor analysis: two latent variables were responsible for the sample distribution within sufficient analysis accuracy, SO4(2-) -NO3- factor (FA1), called as "anthropogenic factor", and Cl- (FA2), called as "marine factor", which corresponding to PC1 and PC2 in orthogonal projection respectively. Results from factor analysis were more practical than from orthogonal projection, that is, the reasons attributed the outlier sample distribution at Zhoushan Archipelago were ascribed into two latent variables: anthropogenic factor (SO4(2-) -NO3-) and marine factor (Cl-). PMID- 16366470 TI - [Eutrophication and pollution level of microcystin in Dianshan Lake]. AB - A survey was conducted in Dianshan Lake to study the eutrophication indexes including total phosphorus (TP), total nitrogen (TN), pH, temperature, diaphaneity and chlorophyll-a level and dominant algae in seasons. The impacts of temperature, light, nitrogen and phosphorus on growth of and microcystin LR production by Microcystis aeraginosa strain under laboratory conditions were studied. Relationship between algal cell density and concentration of microcystin LR were studied. Results suggest that water in Dianshan Lake was eutrophicated. The suitable seasons for algae growth are the end of spring and summer. The annual average of TP and TN were 1.93 mg/L and 0.18 mg/L respectively. And 93.5 and 92.2 percent of TP and TN were higher than the criteria for the third class water body. Significant impact from agriculture was indicated since the peak of algae laged one month after the maxium use of fertilizer. The dominant algae in Dianshan Lake were cyanobacteria, bacillariophyta, cryptophyta and euglenophyta. Microcystis, anabaena and synedra, which excrete toxins and indicate water pollution, and are dominant algae species in summer. M. aeraginosa strain had a biggest growth rate at temperature of 25 degrees C and light intensity of 3 0001x, while microcystin LR production contents reached maximum at 20 degrees C and 5000lx respectively. The optimum TP and TN concentrations for growth of and toxin production by M. aeraginosa were found to be 650 micromol/L and 6.5 micromol/L respectively. TP is suspected to be the limiting factor for the growth of algae both in field and laboratory conditions. Positive correlations between total microcystin LR concentrations and algae cell density or M. aeraginosa cell densities are found. The algae cell density can be used to predict the level of algal toxins in water. PMID- 16366471 TI - [Distribution and species of mercury in water and sediments from Huangpu River]. AB - Levels of total mercury, soluble mercury and particle mercury in water of Huangpu River change greatly, their average values are (0.4 +/- 0.44) ng/mL, (0.27 +/- 0.42)ng/mL, (0.13 +/- 0.10) ng/mL respectively. Mercury in water is mainly in the form of soluble mercury. Average mercury content in sediment of Huangpu River is relative high and up to (204.03 +/- 97.41) ng/g, with a range of 70.52 ng/g - 387.30 ng/g. Mercury content is high in the middle reach of Huangpu River, especially in section of Xidu-Nanpu Bridge, and low in upstream and downstream. Distribution of mercury is hightly related with distribution of industry plants and farming. Locations with high mercury content in sediment are in the downstream of locations with high mercury content in water. Mercury (in sediments) is mainly in the form of residue, exchangeable ions, and humics-bound, seldom is in the form of carbonate-bound. Contrary to residue-bound mercury, exchangeable mercury is low in the middle reach, and high in upstream and downstream. There mainly are residue-bound mercury and little humics-bound mercury, exchangeable mercury, and carbonate-bound mercury in sediment in profile, and the residue-bound mercury increases irregularly with depth. Nearing the Mouth of Yangtze River, mercury in sediment becomes more active. PMID- 16366472 TI - [Effect of environment and nutrient factors on the content of nitrogen and phosphorus in two duckweeds species: Spirodela polyrrhiza and Lemna aequinoctialis]. AB - Effects of temperature, light density, nitrogen level and phosphorus level of medium to the content of two species Spirodela polyrrhiza and Lemna aequinoctialis were researched. The results showed that the content of nitrogen in two species kept constant when the concentration of nitrogen was higher than 3 mg x L(-1) and temperature higher than 25 degrees C. It decreased under lower concentrations of nitrogen and lower temperature. Photon flux density had no effect on it. But weak photon flux density and low temperature led to reduce the content of phosphorus of duckweed. The content of phosphorus in two duckweed species kept constant when photon flux density was higher than 8 000 1x and temperature higher than 25 degrees C. The relationship between the content of phosphorus and concentrations of phosphorus in medium could be described by Monod model. The saturated phosphorus content of Spirodela polyrrhiza was higher than Lemna aequinoctialis. PMID- 16366473 TI - [Property of adsorption about ammonium on suspended mineral matters in eutrophic water]. AB - Static experiments were performed to investigate the adsorption characteristics of ammonium by suspended mineral matters (SPM). Three kinds of suspended mineral matters including montmorillonite, kaolin and the particles separated from the sediments of the Taihu Lake were used as the sorbent. The experiments show that the equation could be gotten within 30 min, and that the ammonium adsorption on mineral particles conformed to Herry isotherm. The parting coefficient of the adsorption about ammonium could reach 548.30, 287.36, 191.27 L/kg in experimental condition that there were 1 000 mg/L mineral particles and 1.0 mg/L ammonium in water with pH 7.00. Some effectors such as temperature, pH and salinity of the water on the adsorption about ammonium were observed. The results show that the quantity of adsorbed ammonium slightly decrease with the increasing of temperature and salinity, and greatly do with increasing of pH. The quantity decrease rapidly also with the increasing of the content of mineral particles below 1 000 mg/L. PMID- 16366474 TI - [Characterization of isolated fractions of NOM from Songhua River]. AB - Natural organic matter (NOM) was concentrated from Songhua River using a reverse osmosis system. The hydrophobic fraction (HyO), hydrophilic fraction (HyI), fulvic acid (FA) and humic acid (HA) were fractionated by the XAD resin, and a commercial humic acid (CHA) was used for comparison. The FA fraction predominant in natural water and account for the large part of DOC, and the aromaticity of these fractions followed the order HA> HyO> FA> HyI. A weak relationship exists between the E4/ E6 and aromaticity. FTIR spectra revealed that the degree of saturation among these fractions was in the order of HyO = FA > HyI > HAM=CHA. Gel chromatograph results showed that the weaker ultraviolet-adsorption fraction of NOM contributed to the most part of molecular weight. Potentiometric titration indicated that the total organic acidity was in the range of 9.84 - 12.15 meq/g, and carboxylic acidity decrease in the order of CHA> HyI > HyO > NOM > FA > HA. 1H NMR spectra demonstrated that NOM and the isolated fractions were predominated by aliphatic functional groups. PMID- 16366475 TI - [Effect of organic acids adsorption on catalytic ozonation with metal oxides]. AB - The effectiveness of ozonation, adsorption and catalytic ozonation with ferric hydroxide (FeOOH) and cerium dioxide (CeO2) as catalysts on removal of several organic acids were investigated. The organic acids selected were benzoic acid (BA), phthalic acid (PA), commercial humic acid (CHA), and humic acid of Songhua River water (SHA) concentrated with reverse osmosis (RO). FeOOH was active in catalytic ozonation of the non-adsorbable BA, with 45% of BA removal higher than ozonation alone. It was inactive in case of ozonation of PA, the adsorption of which on FeOOH caused 5% reduction of its concentration. CeO2 enhanced ozonation of PA that was strongly adsorbed, but not for the non-adsorbed BA. Oxalate ions as product of oxidation of PA were also removed by CeO2 catalytic ozonation. As adsorption of humic acids on CeO2 is stronger than that on FeOOH, its catalytic removal by CeO2 was more than by FeOOH. The activity of catalytic ozonation of humic acids was scavenged in case of competition for adsorption sites on the catalysts by inorganic anions. PMID- 16366476 TI - [O3/H2O2 oxidation processes of cyclops of zooplankton inactivation in water]. AB - The inactivation effects of ozone, hydrogen peroxide, O3/H2O2 processes on Cyclops of zooplankton were discussed. It was found that O3/H2O2 possessed more favorable inactivation effect on Cyclops compared with the others, in distilled water with 1.0 mg/L of O3 and 4 mg/L of H2O2 the inactivation effect was 100% after 30 min contacting; ozone also had favorable inactivation effect, the inactivation effect is 80% by 1.0 mg/L of ozone alone; while hydrogen peroxide didn't, there was almost no inactivation effect by 4 mg/L of H2O2. The best inactivation condition for O3/H2O2 processes, that was, two oxidants were added sequentially with interval in range of 30 to 60 seconds, for them ozone at first. And the influencing factors under various conditions such as the dose of hydrogen peroxide, organic substance content, pH value, etc. were researched and reported. It was found that the effect didn't changed largely with H2O2 on the range of 4 mg/L to 10 mg/L, and inactivation effects on Cyclops of O3/H2O2 were not sensitive to pH value, while it declined obviously as the organic content increased. Finally, synergic removal effect on Cyclops with O3/H2O2 preoxidation followed by clarification process was investigated. The results show that the removal effect on Cyclops is further strengthened by clarification processes after O3/H2O2 pre-oxidation. PMID- 16366477 TI - [Toxicity and influencing factors of liquid chlorine on chironomid larvae]. AB - The excessive propagation of Chironomid larvae (red worm) in the sedimentation tanks is a difficult problem for the normal function of waterworks. The toxic effect of liquid chlorine on the different instar larvae of Chironomid was studied using distilled water as test sample. Furthermore, the effect of pH value, organic matter content, ammonia nitrogen, and algae content on toxicity of liquid chlorine was observed. The results show that the tolerance of Chironomid larvae to liquid chlorine is strengthened with the increase in instar. The 24h semi-lethal concentration (LC50) of liquid chlorine to the 4th instar larvae of Chironomid is 3.39 mg/L. Low pH value and high algae content are helpful to improve the toxic effect of liquid chlorine to Chironomid larvae. In neutral water body, the increase in organic matter content results in the decrease in the death rate of Chironomid larvae. The toxicity of liquid chlorine differs greatly in different concentrations of ammonia nitrogen. The death rate of the 4th instar larvae of Chironomid in raw water is higher by contrast with that in sedimentation tanks water for 24h disposal with various amount of liquid chlorine. PMID- 16366478 TI - [Modeling of inactivation kinetics of E. coli with free chlorine and monochloramine]. AB - The inactivation of Escherichia coli with free chlorine and monochloramine was studied using bench-scale experiments in 0.03 mol/L phosphate buffered saline with pH 7.0 and at 20 degrees C. Inactivation data were fitted to Chick's law, Chick-Watson, Hom, Rational, Hom-Power law, Selleck model. A three-parameter Rational model incorporating a first-order disappearance term for free chlorine residual was found to best describe the observed inactivation of E. coli, and for monochloramine the two-parameter Selleck model was the best fit model. PMID- 16366479 TI - [Characteristics and influencing factors of ammonia contamination from production of monosodium glutamate]. AB - Fluxes of ammonia contamination from production of monosodium glutamate (MSG) were quantitatively observed and relationships with some important influencing factors were statistically analyzed. The study indicates that there is a big difference in the release of ammonia at each step of MSG production including sewage treatment, depending on technical processes. The flux of ammonia released in the process of fermentation was highest, up to 880 mg x (L x min)(-1), and that in the process of refining sugar was lowest, only 0.07 mg x (L x min)(-1). Fluxes of NH3 emission during the whole process of MSG production are significantly related with the concentration of ammonium-nitrogen (NH4+ -N) in discharged solution linearly, and with concentrations of nitrate-nitrogen (NO3- N) including nitrite-nitrogen (NO2- -N) in discharged solution exponentially. Although there is no linear relationship between fluxes of NH3 emission during the whole process of MSG production and pH values in discharged solution, it is significantly related with fluxes of NH3 emission during the process of sewage treatment. PMID- 16366480 TI - [Study on ultrasound-assisted decolorization of methyl orange with Fenton reagent]. AB - Employed with ultrasound, Fenton reagent, ultrasound-Fenton methods, performance and mechanism of decolorization and COD removal were investigated for methyl orange. Results indicated that little decolorization phenomenon was observed with ultrasound solely. For 400mg/L methyl orange solution, 67.51% decolorization ratio and 63.15% COD removal ratio were obtained with Fenton reagent for 30 min. Combined the same operation conditions with ultrasonic field, decolorization ratio and COD removal ratio are 86.91% and 80.32%, respectively. Ultrasound and Fenton reagent have synergistic action on degradation of methyl orange solution. It is oxidative reaction for methyl orange solution with Fenton reagent in strong acidic condition, and it is reduction process for the degradation when pH>5. PMID- 16366481 TI - [Degradation mechanism of acid red B by cathodic oxidation]. AB - Acid red B (ARB) solution with pH = 3 was electrolyzed in a two-chamber cell using Pt/C gas diffusion electrode (GDE) as cathode. The color and COD removal ratio in cathode chamber were 94.2% and 66.8% respectively; and the color and COD removal ratio in anode chamber were 73.3% and 56.6% respectively, which indicated that O2 can be reduced to H2O2 and x OH in the cathode chamber, then ARB was degraded. Through IR and GC-MS analysis for the intermediates of ARB in the cathode chamber, 20 intermediates have been detected, including 14 esters, 3 acids and 3 compounds with -NO2 or N-OH groups and the probable degradation pathway of ARB in the cathode chamber was given. PMID- 16366482 TI - [Preparation of spherical lignin cation adsorption resin with black pulping liquor]. AB - Spherical lignin cation adsorption resin was obtained by two-step method with black liquor of paper mill and characterized by the Scanning electron microscopy (SEM) and Fourier transform infrared spectrometry (FTIR). Firstly, the spherical lignin beads were prepared with black liquor of paper mill by applying reverse phase suspension polymerization technique, and the preparation conditions were also optimized. The suitable conditions include 200 r/min of stirring speed, 90 degrees C of reaction temperature, 1.0 h of reaction time and use epoxy chloropropane (1.5% by weight of lignin) as cross-linking agent, the ideal volume ratio between oil phase and water phase was 3:1 by using kerosene oil as disperse phase, and the perfect dispersant agent was Tween 80 (3% by weight of lignin). Secondly, the spherical lignin cation adsorption resin was developed by grafting the acrylamide onto the back-bone of the spherical lignin beads. The best concentration of acrylamide was 0.72 mol/L at ambient temperature for 2.0 h, and the ideal initiator system was the Fenton reagent of H2O2/Fe2+. Under the above conditions, the ion exchange capacity of the prepared resin could reach 1.640 5 mmol/g. PMID- 16366483 TI - [Photocatalytic degradation of reactive red X-3B by immobilized P25 in a contact oxidation reactor]. AB - The immobilized photocatalyst, P25 supported by glass fiber, was prepared by calcining method. The effect of calcining temperature on the photocatalytic activity and the effect of initial pH and inorganic anions on the photocatalytic degradation of Reactive Red X-3B were explored in a new photocatalytic contact oxidation reactor. The results show that catalyst activity is affected drastically by calcining temperature, the optimal temperature is 300 degrees C; the SEM photos of immobilized catalyst indicated that P25 adhered to glass fiber firmly; The Reactive Red X-3B was more easily degraded in acid solution, when initial pH increasing from 2.0 to 10.0, the degradation rate of Reactive Red X-3B decreased from 90.06% to 42.71%; inorganic anions, CO3(2-) and SO4(2-), had little effect on the degradation of Reactive Red X-3B. However, the degradation of Reactive Red X-3B significantly decreased, 17.03% down, in the presence of Cl . So it was a good alternate scenario to conduct photocatalytic oxidation by using immobilized P25 in a contact oxidation reactor. PMID- 16366484 TI - [Detection of Escherichia coli in wastewater based on enzyme immunoassay]. AB - This research described the fast detection method based on ELISA for the E. coli in drain of wastewater treatment plants. The optimized conditions, such as reaction format (sandwich or direct), the concentration of HRP-E. coli conjugate, anti-HPR antibody and pretreatment of E. coli, were studied. Those results showed that the linear range of detection for E. coli was 10 CFU/ mL - 6 x 10(4) CFU/mL. This method shows the potential ability for the technics control of wastewater treatment plants. Comparing with conventional methods, it is a convenient and sensitive detection method with low cost. PMID- 16366485 TI - [Cell-surface hydrophobicity and degradation characteristics of hydrophobic hydrocarbon degrading bacteria]. AB - Three high efficient hydrocarbon degrading bacteria named HDB-1, HDB-2, HDB-3 were separated from the oil contaminant soil. This paper studied on the cell surface hydrophobicity of these three bacteria and environment influence factor by MATH,and the results show that the xylene-water two-phase system is suitable for studying the cell-surface hydrophobicity of three bacteria; the cell-surface hydrophobicities of HDB-1, HDB-2 and HDB-3 are respectively 68.8%, 57.4% and 64.1%; the hydrophobicity changed with the difference of cultivation and time,carbon source, temperature and pH value; the removal ratio of 1 000 mg/L oil content for 6 day can reach 91.6%, 64.5% and 79.8%. The hydrophobicity is related definitely with the degradation of the organic pollutant, and the degradation rate of hydrophobic organic by the bacteria of high hydrophobicity is quicker than that by the bacteria of low hydrophobicity. PMID- 16366486 TI - [Decolorization of synthetic dyes with a new isolated yeast Candida krusei]. AB - A new yeast isolate Y-G-1, identified as Candida krusei, capable of degrading Reactive Brilliant Red K-2BP, was obtained from soil by screening experiments. This strain gave a maximal decolorization (99%) for Reactive Brilliant Red K-2BP (200 mg/L) after incubation of 12 h. In order to get this decolorization effect, the optimal inoculation volume of Candida krusei was not less than 5%, the optimal pH of culture medium was within a range from 4 to 9, and the concentrations of (NH4)2SO4 and glucose were not lower than 0.1% and 0.5%, respectively. The research results on the mechanism of decolorization show that Candida krusei could biodegrade Reactive Brilliant Red K-2BP. In addition, Candida krusei could also decolorize 62% - 96% of another 9 dyes (50mg/ L). Specially for azo dyes, including Weak Acid Brilliant Red B, Reactive Black KN-B and Reactive Red M-3BE, their decolorization rates were over 90 %, and for triphenylmethane dye, Mordant Blue B, it was up to 93%. All these results indicate that this strain have potential application in dye wastewater treatment. PMID- 16366487 TI - [Research of microbio-remediation of oil-contaminated wetland in Liaodong Bay]. AB - To investigate the effect of microbes on oil degradation, we use the method of introducing selected bacteria to the soil samples. Experiment shows that the combined bacteria are preferable to the single ones because of their synergetic functions. The optimum condition for oil degradation was controlled at pH 8.0, 25 degrees C. Some surfactants are helpful to breed bacteria, thus improves the oil degradation. After adding TW-80 (300 mg x kg(-1)) for 8 days, the number of bacteria was amplified 6.22 times and the oil degradation rate was enlarged 20.1%. Adequate amount of H2O2 is beneficial for microbes to decompose oil, but its overdose may slay bacteria. 400 mg x L(-1) H2O2 each time is suitable dose. In 11 days after 7 000 mg x L(-1) H2O2 was added entirely, the oil degradation rate rose greatly from 39.81% (without H2O2) up to 66.79%. PMID- 16366488 TI - [Culture and biodegradation performance for phenol-degrading bacterium in high phenol concentration]. AB - The activated sludge was acclimated by the inorganic salt medium containing phenol as the sole carbon source. The phenol-degrading bacterium obtained now can degrade completely phenol at initial concentration of 1 700mg/L at 30 degrees C for 15h, and COD can be degraded by 96.9%. Some factors effecting degradation of phenol and COD were studied, which include initial phenol concentration, quantity of bacterium, temperature and pH. Four strains of bacterium were isolated from the mixed phenol-degrading bacterium. It is shown that the degradation effect of mixed bacterium is superior to that of sole strain. PMID- 16366489 TI - [Bioflocculant producing capability by two strains of Bacillus sp. in diversified carbon sources]. AB - Two strains of Bacillus sp. F2 and F6 could produce bioflocculants with high flocculating effects by pure culture and mixed culture. Many kinds of carbon sources could be utilized. Organic compound which molecular weight under 200 such as saccharide, alcoholic aldehyde, ester and organic acid were favorable for producing bioflocculant. And some low-cost biomass scrap could be used as good carbon sources for industrialized production of bioflocculants, such as waste beet molasses, cellulose's fermentation residue and sewage of hydrogen producing reactor. PMID- 16366490 TI - [Capability and stability of degrading rice straw of composite microbial system MC1]. AB - The capability of degrading rice straw of lignocellulolytic composite microbial system MC1 was investigated under different methods of preservation and temperatures treatments of 80 degrees C to 95 degrees C, and stability of composite microbial system MC1 was studied through the method of Denaturing Gradient Gel Electrophoresis (DGGE). The results indicate that the rice straw of 2% dry weight of medium can be degraded completely at 50 degrees C within 10 days under static culture. After 9 days inoculating MC1, the dry weight of rice straw, cellulose, hemicellulose and lignin content was degraded by 81% , 99%, 74% and 51%, respectively. Capability of cellulose degrading and stability of composite microbial system MC1 is sustained under 4 years of continuing subculture, 4 years of dry preservation at room temperature, 4 years of preservation at - 20 degrees C, 1 year of liquid preservation at room temperature and at 4 degrees C, and treatment of 90 degrees C for 30 min, respectively. Plate culture results show that composite microbial system MC1 are consisted of bacteria. The main DNA bands are not changed by the method of 16S rDNA PCR-DGGE after culture of six months so that microbial composition of MC1 is very stable. PMID- 16366491 TI - [Enzymatic hydrolysis of steam-exploded rice straw in membrane bioreactor]. AB - An ultrafiltration membrane bioreactor consisted of 1 - 6 enzymatic vessel was used to increase the reducing sugar concentration. The effects of enzyme loading, the number of enzymatic vessel and the dilution rate on enzymatic hydrolysis of steam-exploded rice straw were studied. The results show that the best condition is: 20FPU/g, four enzymatic vessels and 0.075/h (D). Compared with traditional batch reaction, the conversion of substrate increased significantly from 18% - 21% to 39.5%. Compared with the ultrafiltration membrane bioreactor consisted of one enzymatic vessel, the production of reducing sugar increased from 0.25 g/ g to 0.4g/g, and the final reducing sugar concentration in the product stream increased from 4.56g/L to 27.23g/L. PMID- 16366492 TI - [Experimental research on the anti-wind erosion of typical grasslands]. AB - Wind erosion is one of the most important factors influencing the stability and functions of typical grassland ecosystem and one of the main reasons causing grassland degradation. In order to understand the effects of the human disturbances to the ability of anti-wind erosion of the typical grasslands, a wind tunnel experiment was made and the wind erosion rates (WER) and wind erosion amount (WEA) were selected to illustrate the effects. From the experiment, it can be found that banned-grazing reduced the WEA and the WER markedly. At the same wind speed, the longer the banned-grazing period, the less the WEA and the WER; the WEA and the WER in the grassland cultivation are more than those of the natural grasslands and the differences increase with wind speed. There exists a linear relationship between WER and the disturbance intensity. The results show that grazing and cultivation can weaken the ability of anti-wind erosion of typical grassland ecosystem, and banned-grazing can strengthen the ability. Therefore, in typical grassland areas, taking the active banned-grazing measures is the most effective approach to stop the degradation and trigger the rehabilitation of typical grassland ecosystem. PMID- 16366493 TI - [Influence of PO4(3-) and citrate on REE accumulation and fractionation in wheat]. AB - This paper has studied the influence of phosphate (Pi, one of inorganic ligands) and citrate (Cit, one of organic ligands) on accumulation and fractionation of REEs in wheat based on aqueous culture, added with extraneous mixed REEs (MRE) and ICP-MS analysis technology. The results show that initial phosphate (Pi) solution of different levels followed by exposure to fixed-MRE solution has no significant effects on accumulation of the total concentrations of REEs (sigma REE) in the wheat roots, but it decrease the REE dramatically in the wheat leaves. Simultaneous culture of wheat with mixture of MRE and citrate solution caused obvious decreases of the sigma REE both in wheat roots and leaves. Compared to the control (no Pi or citrate was added), the distribution and fractionation characters of MRE had M-type tetrad effect and MREE enrichment in wheat roots, and W-type tetrad effect and HREE enrichment in wheat leaves. Different levels of Pi had no significant effects on the tetrad effect of MRE, but it notable increased the enrichment of HREE in wheat leaves. Added with citrate of different levels led the fractionation of REE decreasing gradually in wheat roots and leaves, as the concentration of citrate > or = 150 micromol x L( 1), light REE (LREE) enrichment both existed in the roots and leaves. PMID- 16366494 TI - [Influence of arbuscular Mycorrhizal fungi on growth and cu uptake of Elsholtzia splendens]. AB - A pot culture experiment was carried out to study the influence of arbuscular mycorrhizal (AM) fungi on the growth and Cu uptake of E. splendens. Five levels of Cu (0, 50, 100, 200, 400mg x kg(-1)) and two AM fungal inocula, Glomus caledonium 90036 (36) and Acaulospora mellea ZZ (ZZ), and one nonmycorrhizal inoculum (CK), were applied to the soil under sterilized conditions. The plants were harvested after 60 days of growth. Although decreasing with the increase of Cu levels, root infection rates of all the mycorrhizal plants were >50%, which showed that AM fungi colonized E. splendens roots easily. Shoot and root dry matter yields were all increased by two AM inocula at all Cu levels. Shoot Cu concentrations in the plants treated with two AM inocula increased at 200mg x kg( 1) Cu level and below, but did not change significantly at 400mg x kg(-1) Cu level. Root Cu concentrations in the plants treated with two AM inocula did not change at 0 mg x kg(-1) Cu level, but decreased at 50 mg x kg(-1) Cu level and above. In comparison with nonmycorrhizal plants, shoot Cu uptake by mycorrhizal plants all increased at any Cu level, while root Cu uptake increased only at 100 mg x kg(-1) Cu level. In general, AM fungal inoculation can increase Cu translocation from roots to shoots and shoot Cu uptake by E. splendens. PMID- 16366495 TI - [As-hyperaccumulation of Pteris vittata L. as influenced by as concentrations in soils of contaminated fields]. AB - As soil arsenic concentration might have great effect on arsenic accumulation of hyperaccumulator Pteris vittata L. , three sample areas with different As pollution level in the soils are investigated in Chenzhou, Hunan Province, where Pteris vittata L. grows broadly. Great variation of As concentrations is found to be not only among different samples in the same sampling areas but also between rhizosphere and bulk soil of P. vittata L. So it is suggested that rhizosphere soil should be sampled to study the effect of soil contamination on plant accumulation. The As concentration in frond of P. vittata L. increased dramatically at lower As level but decreased slowly when As concentration higher than 1 000 mg/kg. Arsenic translocation factors of P. vittata L. were greater than 1 in all samples in this study, whereas As concentration in shoots in lower As contaminated soil and bioaccumulation factors in higher As contaminated soil are fail to meet the definition of As hyperaccumulator. PMID- 16366496 TI - [Environmental quality of soil polluted by mercury and lead in polymetallic deposit areas of western Hunan Province]. AB - The environmental quality of soil polluted by mercury and lead in typical polymetallic deposit areas of western Hunan province and its natural and human causes were studied. It was found that western Hunan province is a high regional geochemical abnormal region on mercury and lead. The concentrations of mercury and lead in soils in mercury deposit areas were 1 315 and 3.1 times higher than that of the average background of soils in China respectively, while those in soils in lead-zinc deposit areas were 14.8 and 16.1 times higher. Using geoaccumulation index developed by Muller, it was found that either mercury deposit area or lead-zinc deposit area in western Hunan province is polluted by the two elements, the majority soils in mercury deposit areas are seriously polluted by mercury with middle polluted or light polluted by lead, while the majority soils in lead-zinc deposit areas are middle polluted or middle-heavily polluted by lead and middle-heavily polluted by mercury. The sources of metal pollutants in soils in the deposit areas are natural and human causal, the spatial distribution of mercury and lead are dominated by original geochemical situation but seriously affected by human mining activities. PMID- 16366497 TI - [Rapid and high-efficient composting process of municipal sewage sludge]. AB - Based on the static composting process of the municipal sewage sludge (MSS), the paper demonstrates the dynamic composting process of the MSS from Shanghai Quyang Municipal Wastewater Treatment Plant. The dynamics of different operational and evaluating indices during the treatment process are discussed, such as water content, temperature, pH, dissolved organic carbon (DOC) and germination index (GI) and so on. It is concluded that the water content of materials, temperature,DOC and GI can be interpreted as the evaluating indices of the maturation of MSS. Under optimized condition, when the water content of initial materials was adjusted to 60 % +/- 2 % with appropriate sawdust and recycled materials, the results indicated that the temperature of composting materials could reach > 55 degrees C rapidly and last 4 days, thus 100% of the pathogens would be killed out. In the end of composting process, the water content of outputs was reduced exactly to about 35%, DOC could be decomposed by about 50%. T heincompact, odorless, and almost mature output could meet the land application control standard of MSS and the class A standard of US EPA. PMID- 16366498 TI - [Studies of on-site night soil and kitchen garbage treatment]. AB - The biological treatment technique of collection at source and disposition on site of night soil and kitchen garbage were presented. By design project of overall technics, the lab-scale experiments were performed. It was revealed that water consumption of vacuum closestool was about 1 L/time. It consumed 0.4- 0.6L water to shred 1 kg kitchen garbage. Night soil covered 40%, kitchen garbage covered 60% in the influent. Water was controlled at about 93%, the C:N ratio was about 25:1, pH was between 6.2 and 7.3, the optical blend frequency was 6h/d and the overall solid retention time was 28 days in anaerobic digestion reactor. The COD removal rate of mixed supernatant was 91% in anaerobic baffled reactor. It was identified that these phosphorus strains and potassium strains were Bacillus. sp, and biological activated fertilizer was obtained by mixed these strains with digestion sludge which had been dehydrated and deodorized. These strains ability of forming phosphorus and potassium were determined, and the concentration of phosphorus increased 67.5%, potassium increased 33.4%. PMID- 16366499 TI - [Phosphorus characteristics in leachate from soils irrigated with livestock wastewater]. AB - Phosphorus fractions could enter water body from surface runoff and leachate due to excessive irrigation of centralized farm wastewater. Organic P is more mobile than inorganic P in the soil profile and represents a significant proportion of P present in leachate from irrigated farm soils. A set of parallel experiments were conducted to compare the characteristics of organic phosphorus in leachate. The experiment was established in a complete randomized block design with nine replicates. The plots received different combinations of P fertilizer and different rates of pig slurry, i. e. 100, 200,300, 400 t x (hm2 x a)(-1), accordingly, the phosphorus was added to the plots was 6.2, 12.4, 19.2, 24.8 kg x (hm2 x a)(-1), respectively. Leachate was collected over a year period and analyzed for different P fractions. Physico-chemical fractionation of P in leachate indicated that the majority of the P loss from the irrigated soil occurred in unreactive particulate (77% - 90%) P forms. 31P nuclear magnetic resonance analysis of eight leachate samples indicated that unreactive P was mainly comprised of monoester and diester forms of organic P. The presence of phosphomonoesterase and phosphodiesterase activity in leachate resulted in hydrolysis of 9% - 29% of total unreactive P (TUP), indicating that some of the monoesters and diesters can be eventually hydrolyzed into inorganic P forms during P transport. To the treatment P45 + F200, enzyme hydrolysis showed that 33% of the TUP was present as labile monoester P (LMP), followed by 17% as inositol hexakisphosphate (IHP) and 9% as diesters (phospholipids and nucleic acids). The results suggest that LMP, IHP and diesters are an important component of organic P leaching from the irrigated soil. The identification of these organic P forms will improve our understanding of the mechanisms responsible for their release from soils, so that specific mitigation strategies can be implemented at the P source. PMID- 16366500 TI - [Prediction and influence factors of the ramp's noise of the entrance or exit of garages]. AB - Some typical entrances/exits of the underground garages are chosen in urban residential areas. On the basis of the optimization of the positions of the noise sampling points and the groupings of the synchronous sampling points, by means of the acoustical analysis of the noise samples, the relation of the correlative factors, among the ramps' noise of the entrances or exits of the garages, the structure, grade, shape of the ramps, upgrade and downgrade, is studied. The prediction model of the ramp's noise influence of the entrance or exit of the garage is established through amending the noise influence of the entrance or exit of the even concrete road. PMID- 16366501 TI - Human nutritional science on stress control. AB - Severe and prolonged stress exposure impairs homeostatic mechanisms, particularly associated with the onset of depressive illness. The establishment of functional foods that correctly regulate stress response must be an important and new field for human nutrition. This exciting science has been vigorously achieving in the 21st century COE program of University of Tokushima Graduate School. PMID- 16366502 TI - On-line microdevice for stress proteomics. AB - The handling of the cells or tissues is essential for proteomics research or drug screening, where labor is not avoidable. The steps of cell wash, protein extraction, protein denaturing are complicated procedures in conventional method using centrifugation and pipetting in the laboratory. This is the bottle-neck for proteome research. To solve these problems, we propose to utilize the nanotechnology, which will improve the proteomics methodology. Utilizing the nanotechnology, we developed a novel microseparation system, where centrifugation and pipetting are needless. This system has a nanostructured microdevice, by which the cell handling, protein extraction, and antibody assay can be performed. Since cell transfer is needless, all cells are corrected without any loss during the cell-pretreatment procedures, which allowed high reproducibility and enabled the detection of low amount of protein expression. Utilizing the microdevice, we analyzed the stress induced proteins. We further succeeded the screening of food that was useful for immunity and found that an extraction from seaweed promoted the apoptosis of T-lymphoblastic cells. Here, we present an on-line microdevice for stress proteomics. PMID- 16366503 TI - Biomarkers for oxidative stress: measurement, validation, and application. AB - Biomarkers are essential for assessment of oxidative stress and evaluation of antioxidant capacity in vivo. Total hydroxyoctadecadienoic acid (HODE) and 7 hydroxycholesterol measured after reduction and saponification of biological fluids may be used as reliable biomarker. PMID- 16366504 TI - The quest for brain disorder biomarkers. AB - The identification of disease markers in tissues and body fluids requires an extensive and thorough analysis of its protein constituents. In our efforts to identify biomarkers for affective and neurological disorders we are pursuing several different strategies. On one hand we are using animal models that represent defined phenotypes characteristic for the respective disorder in humans. In addition, we are analyzing human specimens from carefully phenotyped patient groups. Several fractions representing different protein classes from human cerebrospinal fluid obtained by lumbar puncture are used for this purpose. Our biomarker identification efforts range from classical proteomics approaches such as two dimensional gel electrophoresis and mass spectrometry to phage display screens with cerebrospinal fluid antibodies. PMID- 16366505 TI - IL-18; a cytokine translates a stress into medical science. AB - Psychological/physical stresses have been reported to exacerbate auto-immune and inflammatory diseases. To clarify a mechanism by which non-inflammatory stresses disrupt host defenses, responses to immobilization stress in mice were investigated, focusing on the role of a multifunctional cytokine, interleukin-18 (IL-18). In the adrenal cortex, the stress induced IL-18 precursor proteins (pro IL-18) via ACTH and a superoxide-mediated caspase-1 activation pathway, resulting in conversion of pro-IL-18 to the mature form which was released into plasma. Inhibitors of caspase-1, reactive oxygen species and P38 MAPK prevented stress induced accumulation of plasma IL-18. These inhibitors also blocked stress induced IL-6 expression. This, together with the observation that IL-6 was not induced in stressed-IL-18 deficient mice, showed that IL-6 induction by stress is dependent on IL-18. In stressed organisms, IL-18 may influence pathological and physiological processes. Controlling the caspase-1 activating pathway to suppress IL-18 levels may provide preventative means against stress-related disruption of host defenses. PMID- 16366506 TI - Approach to novel functional foods for stress control: 1. Toward structure activity relationship and data mining of food compounds by chemoinformatics. AB - Over the past several decades, thousands of natural organic compounds have been isolated from foods, and the information about their structures and biological activities has been accumulated. Despite the long research history of food products and their potential great promise as a medicine and functional food, the systematic research and development have not been carried out extensively. For understanding of the comprehensive structure-activity relationship, we have developed the database system of flavonoids, which are frequently found in vegetables and fruits. We also plan to execute a virtual screening of flavonoids with antidepressant activity by using the database and an advanced chemoinformatic tool. In this section, we will briefly describe (1) flavonoid database and (2) virtual screening of antidepressant compounds. PMID- 16366507 TI - Approach to novel functional foods for stress control 2. Microarray assessment of exercise in healthy volunteers. AB - DNA microarray was used to measure stress response by exercise in peripheral blood leukocytes. Aerobic exercise did not alter mRNA pattern or urinary secretion of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Strenuous exercise increased urinary secretion of 8-OHdG and altered mRNA pattern in microarray. These results suggest that moderate exercise, i. e. aerobic exercise, did not show any change in 8-OHdG, an oxidative stress marker, or mRNA expression in the leukocytes, which might reflect whole body neurohormanal changes. In addition, strenuous exercise produced quite different expression pattern from those of psychological stress. PMID- 16366508 TI - Approach to novel functional foods for stress control 3. Establishment of stress resistant rat model and its mechanism. AB - A stress-resistant rat model was introduced. SPORTS (Spontaneously-Running Tokushima-Shikoku) rats showed significantly shorter time of immobility in the forced swim test compared to control Wister rats. Increase norepinephrine concentration secondary to decreased activity of monoamine oxidase A (MAOA) in hippocampus was observed in this model rats. This model rats are considered to be useful for studying the mechanism of psychological stress. PMID- 16366509 TI - Approach to novel functional foods for stress control 4. Regulation of serotonin transporter by food factors. AB - Serotonin transporters (SERTs) are pre-synaptic proteins specialized for the clearance of serotonin following vesicular release at central nervous system (CNS) and enteric nervous system synapses. SERTs are high affinity targets in vivo for antidepressants such as serotonin selective reuptake inhibitors (SSRIs). These include 'medical' psychopharmacological agents such as analgesics and antihistamines, a plant extract called St John's Wort (Hypericum). Osteoclasts are the primary cells responsible for bone resorption. They arise by the differentiation of osteoclast precursors of the monocyte/macrophage lineage. The expression of SERTs was increased in RANKL-induced osteoclast-like cells. Using RANKL stimulation of RAW264.7 cells as a model system for osteoclast differentiation, we studied the direct effects of food factor on serotonin uptake. The SSRIs (fluoxetine and fluvoxamine) inhibited markedly (approximately 95%) in serotonin transport in differentiated osteoclast cells. The major components of St. John's Wort, hyperforin and hypericine were significantly decreased in serotonin transport activity. Thus, a new in vitro model using RANKL induced osteoclast-like cells may be useful to analyze the regulation of SERT by food factors and SSRIs. PMID- 16366510 TI - Approach to novel functional foods for stress control 5. Antioxidant activity profiles of antidepressant herbs and their active components. AB - Oxidative stress is frequently mentioned in relation to the neurodegenerative diseases. This study examined the effect of three herb extracts, Hypericum perforatum, Ginkgo biloba L. and Apocynum venetum L., and their components on lipid hydroperoxide-induced oxidative stress in PC-12 cells. Among them, the extract of Apocynum venetum and its components showed the remarkable inhibitory effect, indicating that this herb extract serves as a protective agent against lipid peroxidation-related oxidative stress in CNS. Oxidative stress may be associated with the progress of depression, as this extract has been proposed to be an effective antidepresant herb. PMID- 16366511 TI - Stress and nutrition in relation to excess development of chronic disease in Puerto Rican adults living in the Northeastern USA. AB - Although health disparities are well documented among minority populations, they have not been fully explained by socio-economic status. We have demonstrated that Puerto Rican elders in Massachusetts are significantly more likely to have physical disability, depression, cognitive impairment, diabetes and other chronic health conditions than do non-Hispanic white elders living in the same neighborhoods. This suggests that the disparity is not due only to physical or neighborhood location, and that other factors must be influencing these differences. In that study, we also showed that the Puerto Rican elders had diets that were limited in diversity and were relatively low in micronutrient content. In our ongoing cohort study within our Boston Puerto Rican Center for Population Health and Health Disparities, we are investigating the relationships between psychosocial stress, its effect on physiologic burden or "allostatic load" and, in turn, how this is associated with the functional outcomes previously identified as areas of health disparity: depression, cognitive impairment and functional limitation. We further propose that the association between life stress, physiologic response and chronic conditions is modified by nutritional status, with a focus on B vitamins and antioxidant vitamins. PMID- 16366512 TI - Control of oxidative stress and metabolic homeostasis by the suppression of postprandial hyperglycemia. AB - Repeated mental stress may lead to chronic alterations in cortisol and catecholamine concentrations and to insulin resistance. Furthermore, chronically elevated cortisol concentrations may favour the development of abdominal obesity and of the metabolic syndrome. Oxidative stress impairs glucose uptake in muscle and fat and correlates with BMI. Obese subjects with type 2 diabetes, especially soon after the onset of diabetes, usually exhibit postprandial hyperglycemia with delayed hyperinsulinemia. It is recognized that insulin resistance causes postprandial hyperglycemia; however, it is also possible that impairment of early insulin secretion in response to an oral glucose load is the reason why postprandial hyperglycemia occurs. Since even modest increases in postprandial glucose values can be a risk factor for cardiovascular disease. Therefore, the effects of palatinose based functional food which reduces postprandial hyperglycemia and hyperinsulinemia were investigated in rats. This novel food definitely reduced visceral fat accumulation and improved insulin sensitivity. Therefore, it is suggested that functional food which suppresses postprandial glucose level is beneficial for both stress and metabolic controls. PMID- 16366513 TI - Assessment of human stress and depression by DNA microarray analysis. AB - Precise assessment of stress is an imminent issue to deal with stress-related social, medical and psychological problems. Psychological stress is known to stimulate the neuroendocrine, sympathetic nervous, and immune systems. By analyzing mRNA expression levels in leukocytes, which express receptors for hormones, neurotransmitters, growth factors, cytokines, and other stress related signals, levels of stress may be adequately measured. In a series of studies, our group has developed a cDNA microarray specifically designed to measure the mRNA levels of stress-related genes in peripheral blood leukocytes. This microarray enabled us to sensitively detect the response to psychological stress. In addition, our preliminary study suggests that the array could differentiate patients with depression from sex- and age-matched control subjects. PMID- 16366514 TI - Neuroimaging in human dystonia. AB - Functional neuroimaging, such as positron emission tomography (PET) and functional magnetic resonance imaging (fMRI), provides a valuable technique for detecting regional changes in brain metabolic activity associated with human disease. These techniques have been applied in different dystonic disorders including primary generalized dystonia and dopa-responsive dystonia (DRD), as well as focal dystonic syndromes such as torticollis, writer's cramp, and blepharospasm. A common finding is abnormality of the basal ganglia and associated outflow pathways to sensorimotor cortex and other regions involved with motor performance. Other recent imaging research has utilized diffusion based MRI techniques to localize distinct microstructural abnormalities in dystonia patients and gene carriers. This presentation will focus on an integrated approach to understanding the pathophysiology of this genetic and biochemically diverse disorder. PMID- 16366515 TI - Molecular dissection and anatomical basis of dystonia: X-linked recessive dystonia-parkinsonism (DYT3). AB - Pathological findings in dystonia have been unclear. X-linked recessive dystonia parkinsonism (XDP, DYT3), endemic in the Panay island, the Philippines, is characterized by the clinical onset with dystonia followed by parkinsonism. It provides a unique opportunity to explore the anatomical basis of dystonia, because it has discernible pathological changes even at its early phase of dystonia. After extensive searches for the anatomical basis in XDP, we found selective loss of striosomal neurons in the striatum in dystonic patients' brain. Because striosomal neurons inhibit nigrostriatal dopaminergic neurons via GABAergic innervation, the striosomal lesion could account for dopamine excess in the striatum, which in turn causes a hyperkinetic state or dystonia. We also identified the causative gene as one of the general transcription factor genes, TAF1. XDP has certain similarities to Huntington disease not only in pathological and clinical findings, but also the molecular mechanism, which disturbs expression of genes essential for striatal neurons, such as DRD2. Therapeutic intervention may become possible through pharmacological measures that affect gene expression. PMID- 16366517 TI - Construction of a novel database for flavonoids. AB - Flavonoids are polyphenolic compounds that exist ubiquitously in foods of plant origin. Flavonoids show various interesting biological activities, such as removal of oxygen radical, anti-cancer action, improvement of high blood pressure, antibacterial, antibiotic and anti-allergy actions. So far, over 4000 structurally unique flavonoids have been isolated from plant sources. Recently, databases of chemicals have been utilized in to help chemical and biological researches, however the comprehensive database of flavonoids with information about structural, biological and physicochemical properties not yet available. We have constructed the integrated database of flavonoids for nutrition research. PMID- 16366516 TI - Aripiprazole, a novel antipsychotic agent: dopamine D2 receptor partial agonist. AB - It is obvious that DA is an important neurotransmitter in vivo. It is involved in a variety of physiological processes such as mental processes, motor function and hormone regulation. In this context, it is quite understandable that a DA D2 receptor antagonist that inhibits the DA D2 receptor regardless of the state of activity of dopaminergic neurotransmission and inhibit the physiological function of DA can have a variety of adverse effects. In contrast to DA D2 antagonists, aripiprazole acts as an antagonist at the DA D2 receptor in the state of excessive dopaminergic neurotransmission, while it acts as an agonist at the DA D2 receptor in the state of low dopaminergic neurotransmission, and thus attempts to bring the state of dopaminergic neurotransmission to normal. This activity of aripiprazole to regulate dopaminergic neurotransmission is physiologically reasonable, and can be regarded as a stabilizing effect, for which aripiprazole is called a dopamine system stabilizer. PMID- 16366518 TI - Determination of solute lipophilicity by reversed-phase high-performance liquid chromatography (RP-HPLC). AB - Lipophilicity was evaluated by the high-throughput RP-HPLC method. A set of 40 solutes with well-defined solvatochromic parameters were selected in this study. The chromatographic results show that, under the conditions of study, the lipophilicity index log kw was highly correlated with the experimental log Poct. PMID- 16366519 TI - Evaluation of anti-stress nutrients in the endothelial cells with fluorescence indicator. AB - Oxidative stress has emerged as an important pathogenic factor in the development of long-term complications, such as hypertension, atherosclerosis, nephropathy, and cancer. Taking many antioxidants from natural food may be effective to prevent us from those diseases. We have attempted to evaluate the effect of improvement by dietary antioxidants on the endothelial dysfunction induced by hyperglycemia. Fluorescence indicators for reactive oxygen species and nitric oxide were employed to the evaluation. The combination of those fluorescence indicators could be powerful tool to evaluate the effect of anti-stress nutrients on both oxidative stress and endothelial dysfunction. PMID- 16366520 TI - Virtual screening models for finding novel antidepressants. AB - Virtual screening was carried out against various biological targets related to depression by support vector machine classification using the atom-type descriptors. The models were effective as over 75 and 95% of the molecules in external test datasets could be correctly classified, depending on target. Antidepressant compounds had predicted activity against 2.3 targets, on average. An introduction is given to virtual screening and the results of classification experiments are presented. PMID- 16366521 TI - Effects of forced swimming stress on rat brain function. AB - Chronic stress has been reported to be an essential factor for depression. In this study, the effect of forced swimming stress on neurotransmitters and cellular signaling pathway contributing to brain functions was investigated using the forced swimming test (FST) in order to understanding of mechanisms to regulate stress signals in brain. Antidepressant drug, imipramine, significantly reduced the immobility time of male rats in the FST by 85% at a dose of 15 mg/kg for 2 weeks. This result indicated that the swimming stress caused a depressed state in the rats without administration of imipramine. Swimming stress significantly lowered the serotonergic ratio and also markedly enhanced the phosphorylation of ERK1/2 in the hypothalamus region compared to the rats without FST. These phenomena may be included in key mechanisms of the development of depression. PMID- 16366522 TI - Hemodynamic changes in the prefrontal cortex during mental works as measured by multi channel near-infrared spectroscopy (NIRS). AB - To investigate the brain activation in the prefrontal cortex (PFC) during mental works, we examined blood oxygenation changes of healthy subjects by using multi channel near infrared spectropcopy (NIRS). It was directly confirmed that the PFC was activated during mental tasks in vivo and it was suggested that distribution of the activation in the PFC is different among healthy individuals. PMID- 16366523 TI - Oxygen binding to tyrosinase from streptomyces antibioticus studied by laser flash photolysis. AB - Tyrosinases catalyze the o-hydroxylation of monophenols (monophenolase activity) and the oxidation of o-diphenols to o-quinones (diphenolase activity) and possess a dinuclear copper active site. The O2 binding kinetics of oxytyrosinase is studied by flash-photolysis measurements, and the O2 binding rate constant (kO2) is obtained as kO2 = 13 +/- 3 muM-1 s-1. Small molecules, such as carbon monoxide and p-nitrophenol (a substrate-analogue inhibitor), are demonstrated to affect O2 binding kinetics. The activation enthalpy of the rate-limiting step of O2 binding is calculated by the temperature dependence of kO2 to be 12.8 +/- 2.6 kcal/mol. PMID- 16366524 TI - Defining long-range order and local disorder in native alpha-synuclein using residual dipolar couplings. AB - Intrinsically unstructured proteins play key biochemical roles in a vast range of normal and pathological processes. To study these systems, it is necessary to invoke an ensemble of rapidly interconverting conformations. Residual dipolar couplings (RDCs) are particularly powerful probes of the behavior of unfolded proteins, reporting on time and ensemble-averaged conformations up to and beyond the millisecond time scale. In this study, we present a novel interpretation of RDCs in unfolded systems that simultaneously defines long-range structural order and local conformational sampling. This approach is used to describe the structure and dynamics of alpha-Synuclein (alphaS), a protein that is strongly implicated in the development of Parkinson's disease (PD), allowing unambiguous detection of strongly populated conformers containing long-range contacts between the N- and C-terminal domains. The structural model combines two features required for the description of alphaS in solution: local conformational fluctuation based on random sampling of residue-specific phi/psi distributions, and long-range contacts induced by the presence of nonbonding interactions between domains that are distant in primary sequence. Both aspects are found to be necessary for the reproduction of the nonaveraged RDCs from alphaS. Although RDCs have previously been shown to report on local conformational preferences in unstructured proteins, this study shows the additional sensitivity of these measurements to the presence of long-range order in highly flexible systems. PMID- 16366525 TI - Direct measurement of barrier heights in protein folding. AB - A serious limitation in the study of protein folding reactions resides in the lack of experimental methods to measure the absolute height of the free energy barrier. This is particularly unfortunate given that if folding barriers are small, as theory predicts, it might be possible to resolve folding mechanisms directly. Here we explore the performance of a recently developed method to extract folding barriers from equilibrium differential scanning calorimetry (DSC) experiments. To test the method, we compare the thermodynamic barrier heights for 15 proteins obtained from available DSC data with the folding rates measured in kinetic experiments. The correlation between these two parameters is very good (r2 = 0.9) and has a slope consistent with the same energy scale. These results confirm that it is possible to measure free energy barriers for natural proteins from equilibrium DSC experiments. Furthermore, the measured barrier heights are small (<8 RT), in general, and marginal or nonexisting for fast-folding proteins. PMID- 16366526 TI - Entrapping of exohedral metallofullerenes in carbon nanotubes: (CsC60)n@SWNT nano peapods. AB - Exohedral C60-based metallofullerenes, CsC60, have been synthesized and successfully encapsulated into single-wall carbon nanotubes (SWNTs) in high yield by reducing C60 molecules into anions. High-resolution transmission electron microscopy (HRTEM) images and in situ electron energy loss spectroscopy (EELS) indicate that Cs atoms and C60 molecules align within SWNTs as CsC60 exohedral metallofullerenes, and that the formal charge state of encaged CsC60 is expressed as Cs+1C60-1. The present peapods with the exohedral metallofullerenes provide a new insight and the possibility to fine-tune the electronic and transport properties of carbon nanotubes. PMID- 16366527 TI - 1JNH values show that N1...N3 hydrogen bonds are stronger in dsRNA A:U than dsDNA A:T base pairs. AB - Here, we show that 1JNH values are on average 0.4 Hz less negative for double stranded RNA A:U than for DNA A:T base pairs, which, according to existing theory, suggests that RNA N1...N3 hydrogen bond distances are about 0.02 A shorter than those of DNA. Also, there is a statistically relevant trend between 1JNH and 2hDelta13C2 values, which supports the original hypothesis that 2hDelta13C2 values are also sensitive to hydrogen bond distances. Finally, a context dependence is observed for these values, which suggests that hydrogen bonding and base-stacking interactions are coupled. PMID- 16366528 TI - perpendicular organization of macromolecules: synthesis and alignment studies of a soluble poly(iptycene). AB - We describe herein a polymeric material that prefers to align perpendicular to a stretch-aligned polymer host in the solid state. Poly(iptycene) poly-1 was synthesized from monomer 1 under hyperbaric techniques via a Diels-Alder polymerization. Polarized excitation spectra of the anthracene end groups in this material in a stretch-aligned, solution-cast poly(vinyl chloride) (PVC) film showed that the poly(iptycene) prefers to align normal (counter aspect ratio) to the stretching direction of the PVC. This is explained by a "threading" mechanism, whereby the PVC intercalates through the internal free volume presented by poly-1, similar to effects observed in small molecule iptycenes under similar conditions. PMID- 16366529 TI - Nickel-catalyzed cross-coupling reaction of aryl fluorides and chlorides with grignard reagents under nickel/magnesium bimetallic cooperation. AB - Nickel-catalyzed cross-coupling of Grignard reagents with aryl (poly)fluorides or (poly)chlorides can be achieved efficiently in the presence of a new triarylphosphine ligand bearing a nearby hydroxy group. The high reactivity and the unique chemoselectivity (ArF > ArOTf > ArSR) of the catalysis have been attributed to synergy of nickel and magnesium atoms preorganized on the ligand, as has been surmised on the basis of theoretical modeling of the reaction mechanism. PMID- 16366530 TI - Guest-dependent negative thermal expansion in nanoporous prussian blue analogues M(II)Pt(IV)(CN)6.x{H2O} (0 < or = x < or = 2; M = Zn, Cd). AB - The guest-dependent thermal expansion behavior of the nanoporous Prussian Blue analogues MIIPtIV(CN)6.x{H2O} (0 or =100 donors per acceptor. AB - Anthracene derivatives self-assemble into fibers with a high molecular order, as is evidenced by probing the structure with energy-trapping tetracene analogues. Efficient energy transfer processes involving tens to hundreds of donors per acceptor and high emission quantum yields are outstanding characteristics displayed in these self-assembled fibers. PMID- 16366533 TI - Spin-frustrated organic-inorganic hybrids of lindgrenite. AB - We report the syntheses and magnetic properties of hybrid organic-inorganic materials that represent layer-expanded versions of the rare mineral lindgrenite (Cu3(OH)2(MoO4)2). The structures of these compounds feature one-dimensional chains of alternating corner- and edge-sharing Cu(II) triangles. By expanding the inorganic layers of lindgrenite with 4,4'-bipyridine, spin-frustrated antiferromagnetism is revealed by a change of the spin frustration parameter f from 1.2 in lindgrenite to 19.4 in (4,4'-bpy)Cu3(OH)2(MoO4)2. PMID- 16366534 TI - Fast carbon-carbon bond formation by a promiscuous lipase. AB - Lipase B from Candida antarctica was redesigned to catalyze the promiscuous reaction of carbon-carbon bond formation. Mutation of the catalytic serine to alanine afforded a mutant that catalyzed Michael additions of 1,3-dicarbonyls to alpha,beta-unsaturated carbonyl compounds at high specific rates, such as 4000 s 1. The enzyme-catalyzed Michael addition reaction followed saturation kinetics and showed substrate inhibition. The designed enzyme showed high rate enhancements with a catalytic proficiency higher than 108, which is on the same level as that observed for enzymes with native substrates. PMID- 16366536 TI - A fluorobenzene adduct of Ti(IV), and catalytic carboamination to prepare alpha,beta-unsaturated imines and triaryl-substituted quinolines. AB - A rare fluorobenzene adduct of group 4, [(nacnac)Ti=NAr(FC6H5)][B(C6F5)4] (nacnac = [ArNC(tBu)]2CH, Ar = 2,6-iPr2C6H3), has been isolated and structurally characterized. This highly electron-deficient system engages readily in imine metathesis and catalyzes carboamination reactions involving diphenylacetylene and aldimines to afford alpha,beta-unsaturated imines as well as triaryl-substituted quinolines. The latter product results from cyclization of the corresponding electron-rich alpha,beta-unsaturated imine. PMID- 16366535 TI - A reagentless signal-on architecture for electronic, aptamer-based sensors via target-induced strand displacement. AB - Thrombin binding stabilizes the alternative G-quadruplex conformation of the aptamer, liberating the methylene blue (MB)-tagged oligonucleotide to produce a flexible, single-stranded DNA element. This allows the MB tag to collide with the gold electrode surface, producing a readily detectable Faradaic current at thrombin concentrations as low as approximately 3 nM. PMID- 16366537 TI - A chiral luminescent Au16 ring self-assembled from achiral components. AB - A luminescent supramolecular chiral Au16 ring with 4.822 nm perimeter that self assembled from a tetrameric array of achiral Au2 units is described. Intra- and intermolecular Au...Au interactions play an important role in directing its chiral self-assembly. PMID- 16366538 TI - Budding dynamics of multicomponent tubular vesicles. AB - Real-time budding dynamics of multicomponent, tubular lipid vesicles was investigated. By using a fluorescence microscope, three typical growth modes of the buds were observed, corresponding, respectively, to bud growth through coalescence between flat patches, a bud and a patch, as well as that of two buds. The spatial and temporal scales measured in the observation were used to estimate the bending rigidity of the membrane. In the late stage, the continuing coalescence between the buds resulted in large shape deformation of the vesicles, from tubular to spherical vesicles, and the number of the buds decayed with time as N approximately t-2/3. This scaling relation was observed for the first time in experiment and confirmed early theoretical predictions. Our observation showed a difference between the diffusivity of the buds on the lipid membrane and that of the embedded membrane proteins. PMID- 16366539 TI - Metal-organic frameworks with exceptionally high capacity for storage of carbon dioxide at room temperature. AB - Metal-organic frameworks (MOFs) show high CO2 storage capacity at room temperature. Gravimetric CO2 isotherms for MOF-2, MOF-505, Cu3(BTC)2, MOF-74, IRMOFs-11, -3, -6, and -1, and MOF-177 are reported up to 42 bar. Type I isotherms are found in all cases except for MOFs based on Zn4O(O2C)6 clusters, which reveal a sigmoidal isotherm (having a step). The various pressures of the isotherm steps correlate with increasing pore size, which indicates potential for gas separations. The amine functionality of the IRMOF-3 pore shows evidence of relatively increased affinity for CO2. Capacities qualitatively scale with surface area and range from 3.2 mmol/g for MOF-2 to 33.5 mmol/g (320 cm3(STP)/cm3, 147 wt %) for MOF-177, the highest CO2 capacity of any porous material reported. PMID- 16366540 TI - The inverted cucurbit[n]uril family. AB - We report the isolation, characterization, and recognition behavior of iCB[6] and iCB[7], which are diastereomers of CB[6] and CB[7], respectively, containing a single inverted glycoluril unit. Product resubmission experiments establish that these inverted CB[n] are intermediates in the mechanism of CB[n] formation. As a consequence of the inverted glycoluril ring, these inverted cucurbiturils possess a permanent dipole moment, are slightly smaller than their diastereomers, show distinctive selectivity in their recognition behavior, and report directly on the contents of their hydrophobic cavity. PMID- 16366541 TI - Gold(I)-catalyzed stereoselective olefin cyclopropanation. AB - A triphenylphosphinegold(I)-catalyzed cyclopropanation of olefins using propargyl esters as gold(I)-carbene precursors is reported. This reaction provided the basis for the use of a DTBM-SEGPHOS gold(I) complex as a catalyst in the enantioselective (up to 94% ee) preparation of vinyl cyclopropanes with high cis selectivity. PMID- 16366542 TI - Highly selective aerobic oxidation of alcohol catalyzed by a gold(I) complex with an anionic ligand. AB - Highly selective aerobic oxidation of alcohols to carbonyl compounds was catalyzed by a gold(I) complex with an anionic ligand in solution. PMID- 16366543 TI - Air-assisted addition of grignard reagents to olefins. A simple protocol for a three-component coupling process yielding alcohols. AB - Silylmethyl, tertiary-alkyl, alkenyl, and aryl Grignard reagents underwent intermolecular addition to olefins, such as styrenes, conjugated dienes, and enynes under an air atmosphere to give homologated alcohols. For example, (trimethylsilyl)methylmagnesium chloride and alpha-methylstyrene in ether at room temperature under dry air directly furnished 2-phenyl-4-(trimethylsilyl)-2 butanol in good yield. As the Grignard addition to olefins under argon with rigorous exclusion of O2 did not proceed at all, the above reaction should involve a radical mechanism: an alkyl radical generated by the aerial oxidation of the Grignard reagent adds to olefin, which is followed by oxygenation. Representative examples of this transformation, where products were obtained in good to excellent diastereo- or regioselectivity, are also disclosed. PMID- 16366545 TI - Engineering Pt in ceria for a maximum metal-support interaction in catalysis. AB - Conventional supported metal catalysts are metal nanoparticles deposited on high surface area oxide supports with a poorly defined metal-support interface. Typically, the traditionally prepared Pt/ceria catalyzes both methanation (H2/CO to CH4) and water-gas shift (CO/H2O to CO2/H2) reactions. By using simple nanochemistry techniques, we show for the first time that Pt or PtAu metal can be created inside each CeO2 particle with tailored dimensions. The encapsulated metal is shown to interact with the thin CeO2 overlayer in each single particle in an optimum geometry to create a unique interface, giving high activity and excellent selectivity for the water-gas shift reaction, but is totally inert for methanation. Thus, this work clearly demonstrates the significance of nanoengineering of a single catalyst particle by a bottom-up construction approach in modern catalyst design which could enable exploitation of catalyst site differentiation, leading to new catalytic properties. PMID- 16366544 TI - Enzymatic cyclization of dioxidosqualene to heterocyclic triterpenes. AB - Oxidosqualene cyclases normally produce triterpenes from 2,3-(S)-oxidosqualene (OS) but also can cyclize its minor companion (3S,22S)-2,3:22,23-dioxidosqualene (DOS). We explored DOS cyclization in plant triterpene synthesis using a recombinant lupeol synthase (LUP1) heterologously expressed in yeast. Incubation of LUP1 with 3S,22S-DOS gave epoxydammaranes epimeric at C20 and a 17,24 epoxybaccharane in a 4:2:3 ratio. The products reflected a new mechanistic paradigm for DOS cyclization. The structures were determined by NMR and GC-MS, and recent errors in the epoxydammarane literature were rectified. Some DOS metabolites are likely candidates for regulating triterpenoid biosynthesis, while others may be precursors of saponin aglycones. Our in vivo experiments in yeast generated substantial amounts of DOS metabolites in a single enzymatic step, suggesting a seminal role for the DOS shunt pathway in the evolution of saponin synthesis. Quantum mechanical calculations revealed oxonium ion intermediates, whose reactivity altered the usual mechanistic patterns of triterpene synthesis. Further analysis indicated that the side chain of the epoxydammarenyl cation intermediate is in an extended conformation. The overall results establish new roles for DOS in triterpene synthesis and exemplify how organisms can increase the diversity of secondary metabolites without constructing new enzymes. PMID- 16366546 TI - Rhodium-catalyzed reductive aldol reactions using aldehydes as the stoichiometric reductants. AB - Chelated acyl rhodium hydrides, generated from the addition of [Rh(dppe)]ClO4 to beta-sulfide-substituted aldehydes, can function as the stoichiometric reductants in reductive aldol processes. Unsaturated nitriles, esters, and ketones can be used as enolate equivalents, and a variety of simple alpha- and beta-substituted aldehydes can be employed. The use of a second, more electrophilic, aldehyde allows three-component reactions to be performed. PMID- 16366547 TI - Expedient synthesis of cyclopropane alpha-amino acids by the catalytic asymmetric cyclopropanation of alkenes using iodonium ylides derived from methyl nitroacetate. AB - A highly enantioselective (up to 97.5% ee) and diastereoselective (95:5 dr trans/cis) Cu(I)-catalyzed cyclopropanation of alkenes using phenyliodonium ylide generated in situ from iodosobenzene and methyl nitroacetate is reported. The cyclopropanation took place with high enantioselectivity for a wide range of alkenes, and the reaction was performed at room temperature. 1-Nitrocyclopropyl esters are versatile building blocks to access the corresponding cyclopropane amino esters and aminocyclopropanes in two and three steps, respectively, from commercially available products. PMID- 16366548 TI - Allosteric control of oligonucleotide hybridization by metal-induced cyclization. AB - A bis(terpyridine) modified single-stranded DNA smoothly forms a stable cycle by Fe2+-assisted ring closure. A novel, complex type of allosteric behavior is observed with Zn2+, which reversibly off-regulates binding of a complementary oligonucleotide only in a narrow concentration range. PMID- 16366549 TI - Synthesis of multiwalled carbon nanotubes through a modified Wolff-Kishner reduction process. AB - We report the synthesis of highly crystallized multiwalled carbon nanotubes through a modified Wolff-Kishner reduction process at a low temperature of 180 degrees C without adding the conventional catalysts of Fe/Co/Ni into the reaction vessel. The as-synthesized carbon nanotubes are about 10-40 nm in diameter and several tens of micrometers in length. The experimental results indicate that a high concentration of NaOH is essential to the formation of carbon nanotubes. This technique opens a new route for the synthesis of CNTs and other carbon nanostructured materials. PMID- 16366550 TI - A solution to the 2-pyridyl organometallic cross-coupling problem: regioselective catalytic direct arylation of pyridine N-oxides. AB - Direct arylation reactions of pyridine N-oxides occur in excellent yield with complete selectivity for the 2-position with a wide range of aryl bromides. This reactivity permits the use of inexpensive, commercially available, and bench stable pyridine N-oxides as replacements for problematic 2-metallapyridines in palladium-catalyzed cross-coupling reactions. PMID- 16366551 TI - The chlorate-iodine clock reaction. AB - A clock reaction produced by mixing chlorate and iodine solutions in perchloric acid media is reported. This is the first example of a clock reaction using chlorate as a reagent. Increasing chlorate and acid concentration reduces the induction period. Changing the initial iodine concentration does not affect the length of the induction period. The discovery of this clock reaction opens the possibility that a new family of oscillation reactions can be built using chlorate as reagent. PMID- 16366552 TI - Metathesis and metallotropy: a versatile combination for the synthesis of oligoenynes. AB - We have demonstrated that the combined use of enyne metathesis and metallotropic [1,3]-shift of the corresponding alkynyl ruthenium carbenes is a powerful synthetic tool to construct oligoenynes. In this reaction, alkynyl carbene intermediates formed from an initial ring-closing metathesis reaction (RCM) undergo repetitive [1,3]-shifts and RCMs to give the final products. Linear poly 1,3-diynes containing repeating functionality of the type -[XCH2CCCCCH2]n- generated long-chain conjugated oligoenynes up to n = 5. PMID- 16366553 TI - Synthesis and characterization of stabilized subnanometric cobalt metal particles. AB - Subnanometric cobalt metallic particles, with an average size of 0.8 nm and an estimated number of 50 atoms, have been stabilized in the confined spaces within the nanopores of crystalline molecular sieves. Remarkably, these clusters show a rapid vanishing of the magnetization as the temperature is increased from 10 to 20 K because of the ferromagnetic-paramagnetic transition together with thermal fluctuations of the remaining moment. This dramatic reduction of the transition temperature is due to strong finite size effects. Such behavior, predicted for very small metallic particles, was never observed before due to the inherent difficulty in achieving subnanometric stable metallic particles. PMID- 16366554 TI - Hydroxylation of phenolic compounds by a peroxodicopper(II) complex: further insight into the mechanism of tyrosinase. AB - The dicopper(I) complex [Cu2(MeL66)]2+ (where MeL66 is the hexadentate ligand 3,5 bis-{bis-[2-(1-methyl-1H-benzimidazol-2-yl)-ethyl]-amino}-meth ylbenzene) reacts reversibly with dioxygen at low temperature to form a mu-peroxo adduct. Kinetic studies of O2 binding carried out in acetone in the temperature range from -80 to -55 degrees C yielded the activation parameters DeltaH1(not equal) = 40.4 +/- 2.2 kJ mol(-1), DeltaS1)(not equal) = -41.4 +/- 10.8 J K(-1) mol(-1) and DeltaH( 1)(not equal) = 72.5 +/- 2.4 kJ mol(-1), DeltaS(-1)(not equal) = 46.7 +/- 11.1 J K(-1) mol(-1) for the forward and reverse reaction, respectively, and the binding parameters of O2 DeltaH degrees = -32.2 +/- 2.2 kJ mol(-1) and DeltaS degrees = 88.1 +/- 10.7 J K(-1) mol(-1). The hydroxylation of a series of p-substituted phenolate salts by [Cu2(MeL66)O2]2+ studied in acetone at -55 degrees C indicates that the reaction occurs with an electrophilic aromatic substitution mechanism, with a Hammett constant rho = -1.84. The temperature dependence of the phenol hydroxylation was studied between -84 and -70 degrees C for a range of sodium p cyanophenolate concentrations. The rate plots were hyperbolic and enabled to derive the activation parameters for the monophenolase reaction DeltaH(not equal)ox = 29.1 +/- 3.0 kJ mol(-1), DeltaS(not equal)ox = -115 +/- 15 J K(-1) mol(-1), and the binding parameters of the phenolate to the mu-peroxo species DeltaH degrees(b) = -8.1 +/- 1.2 kJ mol(-1) and DeltaS degrees(b) = -8.9 +/- 6.2 J K(-1) mol(-1). Thus, the complete set of kinetic and thermodynamic parameters for the two separate steps of O2 binding and phenol hydroxylation have been obtained for [Cu2(MeL66)]2+. PMID- 16366555 TI - Intramolecular metathesis of a vinyl group with vinylidene C=C double bond in Ru complexes. AB - The cationic complex {[Ru]=C=CHCPh2CH2CH=CH2}BF4 (3a, [Ru] = (eta5 C5H5)(PPh3)2Ru) in solution transforms to {[Ru]=C=CHCH2CPh2CH=CH2}BF4 (4a) via a new metathesis process of the terminal vinyl group with the C=C of the vinylidene group which is confirmed by 13C labeling studies. This transformation is irreversible as revealed by deuteration and decomplexation studies. The cationic complex {[Ru]=C=CHCPh2CH2CMe=CH2}BF4 (3b) undergoes a cyclization process yielding 6b containing a eta2-cyclic allene ligand which is fully characterized by single-crystal X-ray diffraction analysis. Analogous complexes 4a' and 6b' ([Ru] = (eta5-C5H5)(dppe)Ru) containing dppe ligands were similarly obtained from protonation of the corresponding acetylide complexes via formation of vinylidene intermediate. Protonation of the acetylide complex containing a terminal alkynyl group [Ru]-CCCPh2CH2CCH (2c) generates the vinylidene complex {[Ru]=C=CHCPh2CH2CCH}BF4 (3c) which again undergoes an irreversible transformation to give {[Ru]=C=CHCH2CPh2CCH}BF4 (4c) possibly via a pi coordinated alkynyl complex followed by hydrogen and metal migration. No similar transformation is observed for the analogous dppe complex 3c'. With an extra methylene group, complex {[Ru]=C=CHCPh2CH2CH2CH=CH2}BF4 (3d) and complex {[Ru]=C=CHCPh2CH2Ph}BF4 (3e) are stable. The presence of a gem-diphenylmethylene moiety at the vinylidene ligand with the appropriate terminal vinyl or alkynyl group along with the correct steric environment implements such a novel reactivity in the ruthenium vinylidene complexes. PMID- 16366556 TI - Aza-cope rearrangement-mannich cyclizations for the formation of complex tricyclic amines: stereocontrolled total synthesis of (+/-)-gelsemine. AB - A detailed examination of the use of aza-Cope rearrangement-Mannich cyclization sequences for assembling the azatricyclo[4.4.0.0(2,8)]decane core of gelsemine is described. Iminium ions and N-acyloxyiminium ions derived from endo-oriented 1 methoxy- or 1-hydroxybicyclo[2.2.2]oct-5-enylamines do not undergo the first step of this sequence, cationic aza-Cope rearrangement, to form cis hydroisoquinolinium ions. However, the analogous base-promoted oxy-aza-Cope rearrangement does take place to form cis-hydroisoquinolones containing functionality that allows iminium ions or N-acyloxyiminium ions to be generated regioselectively in a subsequent step. Mannich cyclization of cis hydroisoquinolones prepared in this way efficiently assembles the azatricyclo[4.4.0.0(2,8)]decane unit of gelsemine. Using a sequential base promoted oxy-aza-Cope rearrangement/Mannich cyclization sequence, gram quantities of azatricyclo[4.4.0.0(2,8)]decanone 18, a central intermediate in our total of (+/-)-gelsemine, were prepared from 3-methylanisole in 12 steps and 16% overall yield. PMID- 16366557 TI - Use of the intramolecular Heck reaction for forming congested quaternary carbon stereocenters. Stereocontrolled total synthesis of (+/-)-gelsemine. AB - Intramolecular Heck reactions of alpha,beta-unsaturated 2-haloanilides derived from azatricyclo[4.4.0.0(2,8)]decanone 5 efficiently install the congested spirooxindole functionality of gelsemine. Depending upon the Heck reaction conditions and the nature of the beta-substituent, either products having the natural or unnatural configuration of the spirooxindole group are formed predominantly. Efforts to elaborate the hydropyran ring of gelsemine from the endo-oriented nitrile substituent of pentacyclic Heck product 18 were unsuccessful. Important steps in the ultimately successful route to (+/-) gelsemine (1) are as follows: (a) intramolecular Heck reaction of tricyclic beta methoxy alpha,beta-unsaturated 2-iodoanilide 68 in the presence of silver phosphate to form pentacyclic product 69 having the unnatural configuration of the spirooxindole fragment, (b) formation of hexacyclic aziridine 80 from the reaction of cyanide with intermediate 79 containing an N-methoxycarbonyl-beta bromoethylamine fragment, (c) introduction of C17 by ring-opening of the aziridinium ion derived from aziridine 80, and (d) base-promoted skeletal rearrangement of pentacyclic equatorial alcohol 82 to form the oxacyclic ring and invert the spirooxindole functional group to provide hexacyclic gelsemine precursor 83. PMID- 16366558 TI - Sigma-borane coordinated to nickel(0) and some related nickel(II) trihydride complexes. AB - The reactions of the complexes [(dcype)NiH]2, 1, [(dippe)NiH]2, 2, and [(dtbpe)NiH]2, 3, with a mixture of BEt3 and Super-Hydride (LiHBEt3) afforded sigma-borane nickel(0) compounds of the type [(dcype)Ni(sigma-HBEt2)], 4, [(dippe)Ni(sigma-HBEt2)], 5, [(dtbpe)Ni(sigma-HBEt2)], 6, respectively, with the concomitant formation in each case of [(dcype)2Ni2)(H)3][BEt4], 7, [(dippe)2Ni2(H)3][BEt4], 8 and [(dtbpe)2Ni2(H)3][BEt4], 9, respectively. X-ray crystal structures are reported for 4 and 8. The reaction of BEt3 and LiHBEt3 was also reviewed in detail. PMID- 16366559 TI - C-H activations at iridium(I) square-planar complexes promoted by a fifth ligand. AB - In the presence of ligands such as acetonitrile, ethylene, or propylene, the Ir(I) complex [Ir(1,2,5,6-eta-C8H12)(NCMe)(PMe3)]BF4 (1) transforms into the Ir(III) derivatives [Ir(1-kappa-4,5,6-eta-C8H12)(NCMe)(L)(PMe3)]BF4 (L = NCMe, 2; eta2-C2H4, 3; eta2-C3H6, 4), respectively, through a sequence of C-H oxidative addition and insertion elementary steps. The rate of this transformation depends on the nature of L and, in the case of NCMe, the pseudo-first-order rate constants display a dependence upon ligand concentration suggesting the formation of five-coordinate reaction intermediates. A similar reaction between 1 and vinyl acetate affords the Ir(III) complex [Ir(1-kappa-4,5,6-eta-C8H12){kappa-O-eta2 OC(Me)OC2H3}(PMe3)]BF4 (7) via the isolable five-coordinate Ir(I) compound [Ir(1,2,5,6-eta-C8H12){kappa-O-eta2-OC(Me)OC2H3}(PMe3)]BF4 (6). DFT (B3LYP) calculations in model complexes show that reactions initiated by acetonitrile or ethylene five-coordinate adducts involve C-H oxidative addition transition states of lower energy than that found in the absence of these ligands. Key species in these ligand-assisted transformations are the distorted (nonsquare-planar) intermediates preceding the intramolecular C-H oxidative addition step, which are generated after release of one cyclooctadiene double bond from the five coordinate species. The feasibility of this mechanism is also investigated for complexes [IrCl(L)(PiPr3)2] (L = eta2-C2H4, 27; eta2-C3H6, 28). In the presence of NCMe, these complexes afford the C-H activation products [IrClH(CH=CHR)(NCMe)(PiPr3)2] (R = H, 29; Me, 30) via the common cyclometalated intermediate [IrClH{kappa-P,C-P(iPr)2CH(CH3)CH2}(NCMe)(PiPr3)] (31). The most effective C-H oxidative addition mechanism seems to involve three-coordinate intermediates generated by photochemical release of the alkene ligand. However, in the absence of light, the reaction rates display dependences upon NCMe concentration again indicating the intermediacy of five-coordinate acetonitrile adducts. PMID- 16366560 TI - Regioselectively trisilylated hexopyranosides through homogeneously catalyzed silane alcoholysis. AB - The iridium complex [Ir(COD)(PPh3)2]+ SbF6- reacts with tert-butyldimethylsilane in DMA to form [IrH2(Sol)2(PPh3)2]+ SbF6-, which is an active catalyst for the regioselective di- and trisilylation of a series of representative methyl hexopyranosides, beta-1,6-anhydrohexopyranosides and 1,3,5-O-methylidene inositol. The corresponding 2,3,6- and 2,4,6-silylated glycosides are obtained in a separable mixture of 47-89% (2,3,6-isomers) and 9-25% (2,4,6-isomers) yield in a single-pot reaction. The 2,4-disilylated derivatives of mannosan, galactosan, and 1,3,5-O-methylidene inositol as well as persilylated levoglucosan are accessible in >85% yield by this method. The homogeneous nature of the catalysts is a prerequisite for the effective di-/trisilylation, as nanoparticle colloid catalysts generated in situ from Pd2(dba)3 (approximately 1.5 nm average particle size) or Ru2Cl5(MeCN)7 (approximately 0.65 nm average particle size) result in only low yields. PMID- 16366561 TI - Mechanistic impact of water addition to SmI2: consequences in the ground and transition state. AB - The mechanistic impact of water addition to SmI2 on the ground state and rate limiting transition state structures in the reduction of benzyl bromide was determined using UV-vis spectroscopy, cyclic voltammetry, vapor pressure osmommetry, and stopped-flow spectrophotometric studies. The results obtained from these studies show that, upon addition of water, SmI2 in THF (or DME) becomes partially water-solvated by displacing metal-coordinated solvent. Further addition of water displaces remaining bound solvent and induces a monomer-dimer equilibrium of the SmI2-water complex. Concomitant with this process, a thermodynamically more powerful reductant is created. Rate studies on the reduction of benzyl bromide by SmI2-water are consistent with reaction occurring through a dimeric transition state with the assembly of the activated complex requiring an equivalent of water at low concentrations but not at higher concentrations. The mechanistic complexity of the SmI2-water system shows that simple empirical models describing the role of water in SmI2-mediated reductions are likely to contain a high degree of uncertainty. PMID- 16366563 TI - Pressure independence of stereomutation and fragmentation in the bis spirocyclobutanes, anti- and syn-2,9-dicyanodispiro[5.0.5.2]tetradeca-1,8-dienes. AB - Pressure dependent rate constants and volumes of activation for stereomutational interconversions of the cyclobutanes, anti- and syn-2,9 dicyanodispiro[5.0.5.2]tetradeca-1,8-dienes (anti-6 and syn-6), and for their fragmentation to 1-cyano-3-methylenecyclohexene (5) have been determined. Although fragmentations might have been expected to have larger and thus more positive volumes of activation than stereomutation, both processes have essentially identical volumes of activation within experimental uncertainties at 50.1 degrees C over the pressure range, 1-3000 bar: DeltaV(not equal) = (+7.4 to +9.9) +/- 2.0 cm3 mol(-1). While these positive values are consistent with the rate of entry into the hypothetical caldera being determined by breaking the weakest bond in the cyclobutanes, the insensitivity of the product-determining exit channels argues against the obligatory second bond-breaking being a significant factor in fragmentation. PMID- 16366562 TI - Structural evolution of the chromophore in the primary stages of trans/cis isomerization in photoactive yellow protein. AB - We have studied the structural changes induced by optical excitation of the chromophore in wild-type photoactive yellow protein (PYP) in liquid solution with a combined approach of polarization-sensitive ultrafast infrared spectroscopy and density functional theory calculations. We identify the nuC8-C9 marker modes for solution phase PYP in the P and I0 states, from which we derive that the first intermediate state I0 that appears with a 3 ps time constant can be characterized to have a cis geometry. This is the first unequivocal demonstration that the formation of I0 correlates with the conversion from the trans to the cis state. For the P and I0 states we compare the experimentally measured vibrational band patterns and anisotropies with calculations and find that for both trans and cis configurations the planarity of the chromophore has a strong influence. The C7=C8 (C9=O)-S moiety of the chromophore in the dark P state has a trans geometry with the C=O group slightly tilted out-of-plane, in accordance with the earlier reported structure obtained in an X-ray diffraction study of PYP crystals. In the case of I0, experiment and theory are only in agreement when the C7=C8-(C9=O)-S moiety has a planar configuration. We find that the carboxylic side group of Glu46 that is hydrogen-bonded to the chromophore phenolate oxygen does not alter its orientation on going from the electronic ground P state, via the electronic excited P state to the intermediate I0 state, providing conclusive experimental evidence that the primary stages of PYP photoisomerization involve flipping of the enone thioester linkage without significant relocation of the phenolate moiety. PMID- 16366564 TI - Carbocation-forming reactions in ionic liquids. AB - A number of trifluoroacetates, mesylates, and triflates have been studied in ionic liquids. Several lines of evidence indicate that all of these substrates react via ionization to give carbocationic intermediates. For example, cumyl trifluoroacetates give mainly the elimination products, but the Hammett rho+ value of -3.74 is consistent with a carbocationic process. The analogous exo-2 phenyl-endo-3-deutero-endo-bicyclo[2.2.1]hept-2-yl trifluoroacetate gives an elimination where loss of the exo-hydrogen occurs from a cationic intermediate. 1 Adamantyl mesylate and 2-adamantyl triflate react to give simple substitution products derived from capture of 1- and 2-adamantyl carbocations by the residual water in the ionic liquid. The triflate derivative of pivaloin, trans-2 phenylcyclopropylcarbinyl mesylate, 2,2-dimethoxycyclobutyl triflate, the mesylate derivative of diethyl (phenylhydroxymethyl)-thiophosphonate, and Z-1 phenyl-5-trimethylsilyl-3-penten-1-yl trifluoroacetate all give products derived carbocation rearrangements (kDelta processes). anti-7-Norbornenyl mesylate gives products with complete retention of configuration, indicative of involvement of the delocalized 7-norbornenyl cation. 1,6-Methano[10]annulen-11-yl triflate reacts in [BMIM][NTf2] to give 1,6-methano[10]annulen-11-ol, along with naphthalene, an oxidized product derived from loss of trifluoromethanesulfinate ion. Analogous loss of CF3SO2- can be seen in reaction of PhCH(CF3)OTf. Ionic liquids are therefore viable solvents for formation of carbocationic intermediates via kC and kDelta processes. PMID- 16366565 TI - A stereoelectronic effect on turn formation due to proline substitution in elastin-mimetic polypeptides. AB - Stereoelectronic effects have been identified as contributing factors to the conformational stability of collagen-mimetic peptide sequences. To assess the relevance of these factors within other protein structural contexts, three polypeptide sequences were prepared in which the sequences were derived from the canonical repeat unit (Val-Pro-Gly-Val-Gly) of the protein material elastin. These elastin-mimetic polypeptides, elastin-1, elastin-2, and elastin-3, incorporate (2S)-proline, (2S,4S)-4-fluoroproline, and (2S,4R)-4-fluoroproline, respectively, at the second position of the elastin repeat. Calorimetric and spectroscopic investigations of these three polypeptides indicate that the incorporation of the substituted proline residues had a dramatic effect upon the self-assembly of the corresponding elastin peptide. The presence of (2S,4R)-4 fluoroproline in elastin-3 lowered the temperature of the phase transition and increased the type II beta-turn population with respect to the parent polypeptide, while the presence of (2S,4S)-4-fluoroproline in elastin-2 had the opposite effect. These results suggest that stereoelectronic effects could either enhance or hinder the self-assembly of elastin-mimetic polypeptides, depending on the influence of the proline analogue on the energetics of the beta-turn conformation that develops within the pentapeptide structural repeats above the phase transition. Density functional theory (DFT) was employed to model three possible turn types (betaI-, betaII-, and inverse gamma-turns) derived from model peptide segments (MeCO-Xaa-Gly-NHMe) (Xaa = Pro, 4S-F-Pro, or 4R-F-Pro) corresponding to the turn-forming residues of the elastin repeat unit (Val-Pro Gly-Val-Gly). The results of the these calculations suggested a similar outcome to the experimental data for the elastin-mimetic polypeptides, in that type II beta-turn structures were stabilized for peptide segments containing (2S,4R) fluoroproline and destabilized for segments containing (2S,4S)-fluoroproline relative to the canonical proline residue. PMID- 16366566 TI - A quadruply hydrogen bonded heterocomplex displaying high-fidelity recognition. AB - An exceptionally strong quadruply hydrogen-bonded complex is formed between 2,7 diamido-1,8-naphthyridine 3 (DAN) and the butylurea of guanosine 6 (UG) in chloroform. The UG unit can be prepared in four steps from guanosine on a 10 g scale in excellent yields without chromatographic purification. The association constant (Kassoc approximately 5 x 10(7) M(-1)) for the UG.DAN complex determined by fluorescence energy transfer from the naphthyridine unit of 3 to coumarin 343 covalently linked UG (18) is among the highest reported for a neutral DNA base pair analogue. The weak self-association of DAN (Kdimer < 10 M(-1)) and UG (Kdimer ca. 200-300 M(-1)) means that the UG.DAN complex forms with unparalleled fidelity. PMID- 16366567 TI - Base pairing and replicative processing of the formamidopyrimidine-dG DNA lesion. AB - The 2,6-diamino-4-hydroxy-5-formamidopyrimidine of 2'-deoxyguanosine (FaPydG) is one of the major DNA lesions found after oxidative stress in cells. To clarify the base pairing and coding potential of this major DNA lesion with the aim to estimate its mutagenic effect, we prepared oligonucleotides containing a cyclopentane based analogue of the DNA lesion (cFaPydG). In addition, oligonucleotides containing the cyclopentane analogue of 2'-deoxyguanosine (cdG), and oligonucleotides containing 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) were synthesized. The thermodynamic stability of duplexes containing these building blocks and all canonical counterbases were determined by concentration dependent melting-point measurements (van't Hoff plots). The data reveal that cFaPydG greatly destabilizes a DNA duplex (DeltaDeltaG degrees (298K) approximately 2-4 kcal mol(-1)). The optimal base pairing partner for the cFaPydG lesion is dC. Investigation of duplexes containing dG and cdG shows that the effect of substituting the deoxyribose by a cyclopentane moiety is marginal. The data also provide strong evidence that the FaPydG lesion is unable to form a base pair with dA. Our computational studies indicate that the syn-conformation required for base pairing with dA is energetically unfavorable. This is in contrast to 8-oxodG for which the syn-conformation represents the energetic minimum. Kinetic primer extension studies using S. cerevisiae Pol eta reveal that cFaPydG is replicated in an error-free fashion. dC is inserted 2-3 orders of magnitude more efficiently than dT or dA, showing that FaPydG is a lesion which retains the coding potential of dG. This is also in contrast to 8-oxodG, for which base pairing with dC and dA was established. PMID- 16366568 TI - Electronically coupled MM quadruply-bonded complexes (M = Mo or W) employing functionalized terephthalate bridges: toward molecular rheostats and switches. AB - Toluene solutions of M2(O2C(t)Bu)4 (M = Mo, W; 2 equiv) react with a range of functionalized terephthalic acids, HO2CArCO2H (Ar = C6H4, C6F4, C6Cl4, C6H2-2,5 Cl2, C6H2-2,5-(OH)2, C6H3-2-F), to give [(tBuCO2)3M2]2[mu-O2CArCO2]. These compounds show intense ML(bridge)CT absorptions in the visible region of the electronic spectrum, and the terephthalate bridge serves to electronically couple the two M2 units via interactions between the M2 delta and bridge pi orbitals. Electronic structure calculations reveal how the degree of electronic coupling is controlled by the dihedral angles between the terephthalate C6 ring and the two CO2 units and the degree of interaction between the M4 delta MOs and the LUMO of the bridge. Both of these factors are controlled by the aryl substituents, and collectively these determine the thermochromism displayed by these complexes in solution together with the physical properties of the oxidized radical cations as determined by electrochemical studies (CV, DPV), UV-vis-NIR and EPR spectroscopic methods. PMID- 16366569 TI - Pressure enhanced conductivity in bis-1,2,3-thiaselenazolyl dimers. AB - A synthetic sequence to salts of N-alkylated pyridine-bridged 1,2,3 thiaselenazolo-1,2,3-thiaselenazolylium cations [2]+ (R1 = Me, Et; R2 = H) is described. The corresponding radicals 2 (R1 = Me, Et; R2 = H) can be generated from the cations by chemical or electrochemical reduction. Crystals of the two radicals are isostructural and consist of interpenetrating pi-stacked arrays of closed-shell Se-Se sigma-bonded dimers [2]2 laced together with numerous short intermolecular Se- - -Se, Se- - -S, and Se- - -N contacts. Variable-temperature magnetic, conductivity, and near-infrared optical measurements indicate that the bulk materials behave as small band gap semiconductors with room-temperature conductivities sigma(RT) near 10(-6) S cm(-1) and thermal activation energies Ea of 0.32 eV (R1 = Me) and 0.36 eV (R1 = Et). LMTO band structure calculations on both compounds are consistent with this interpretation. The application of external pressure leads to dramatic increases in conductivity; at 4 GPa sigma(RT) reaches a value near 10(-1) S cm(-1) for R1 = Me and 10(-2) S/cm for R1 = Et. The conductivity remains activated for both compounds, but for R1 = Me the activation energy Ea is reduced to near 0.03 eV at 5 GPa, suggestive of a weakly metallic state. PMID- 16366570 TI - Conformational dynamics of the isoalloxazine in substrate-free p-hydroxybenzoate hydroxylase: single-molecule studies. AB - p-Hydroxybenzoate hydroxylase (PHBH) is a homodimeric enzyme in which each subunit noncovalently binds one molecule of FAD in the active site. PHBH is a model system for how flavoenzymes regulate reactions with oxygen. We report single-molecule fluorescence studies of PHBH in the absence of substrate that provide data consistent with the hypothesis that a critical step in substrate binding is the movement of the isoalloxazine between an "in" conformation and a more exposed or "open" conformation. The isoalloxazine is observed to move between these conformations in the absence of substrate. Studies with the Y222A mutant form of PHBH suggest that the exposed conformation is fluorescent while the in-conformation is quenched. Finally, we note that many of the single molecule-fluorescence trajectories reveal a conformational heterogeneity, with populations of the enzyme characterized by either fast or slow switching between the in- and open-conformations. Our data also allow us to hypothesize a model in which one flavin in the dimer inhibits the motion of the other. PMID- 16366571 TI - Electrochemical definitions of O2 sensitivity and oxidative inactivation in hydrogenases. AB - A new strategy is described for comparing, quantitatively, the ability of hydrogenases to tolerate exposure to O2 and anoxic oxidizing conditions. Using protein film voltammetry, the inherent sensitivities to these challenges (thermodynamic potentials and rates of reactions) have been measured for enzymes from a range of mesophilic microorganisms. In the absence of O2, all the hydrogenases undergo reversible inactivation at various potentials above that of the H+/H2 redox couple, and H2 oxidation activities are thus limited to characteristic "potential windows". Reactions with O2 vary greatly; the [FeFe] hydrogenase from Desulfovibrio desulfuricans ATCC 7757, an anaerobe, is irreversibly damaged by O2, surviving only if exposed to O2 in the anaerobically oxidized state (which therefore affords protection). In contrast, the membrane bound [NiFe]-hydrogenase from the aerobe, Ralstonia eutropha, reacts reversibly with O2 even during turnover and continues to catalyze H2 oxidation in the presence of O2. PMID- 16366572 TI - Quantitative analysis of backbone dynamics in a crystalline protein from nitrogen 15 spin-lattice relaxation. AB - A detailed analysis of nitrogen-15 longitudinal relaxation times in microcrystalline proteins is presented. A theoretical model to quantitatively interpret relaxation times is developed in terms of motional amplitude and characteristic time scale. Different averaging schemes are examined in order to propose an analysis of relaxation curves that takes into account the specificity of MAS experiments. In particular, it is shown that magic angle spinning averages the relaxation rate experienced by a single spin over one rotor period, resulting in individual relaxation curves that are dependent on the orientation of their corresponding carousel with respect to the rotor axis. Powder averaging thus leads to a nonexponential behavior in the observed decay curves. We extract dynamic information from experimental decay curves, using a diffusion in a cone model. We apply this study to the analysis of spin-lattice relaxation rates of the microcrystalline protein Crh at two different fields and determine differential dynamic parameters for several residues in the protein. PMID- 16366573 TI - Utilization of a combination of weak hydrogen-bonding interactions and phase segregation to yield highly thermosensitive supramolecular polymers. AB - Supramolecular polymerization, i.e., the self-assembly of polymer-like materials through the utilization of the noncovalent bond, is a developing area of research. In this paper, we report the synthesis and investigation of nucleobase terminated (N6-anisoyl-adenine and N4-(4-tert-butylbenzoyl)cytosine) low molecular weight poly(THF) macromonomers (<2000 g mol(-1)). Even though the degree of interaction between the nucleobase derivatives is very low (<5 M(-1)) these macromonomers self-assemble in the solid state to yield materials with film and fiber-forming capability. While the mechanical properties of films of both materials show extreme temperature sensitivity, resulting in the formation of very low viscosity melts, they do behave differently, which is attributed to the nature of the self-assembly controlled by the nucleobase. A combination of FT-IR, WAXD, and rheological experiments was carried out to further investigate the nature of the self-assembly in these systems. The studies demonstrate that a combination of phase segregation between the hard nucleobase chain ends and the soft poly(THF) core combined with aromatic amide hydrogen bonding is utilized to yield the highly thermosensitive supramolecular polymeric materials. In addition, analysis of the data suggests that the rheological properties of these supramolecular materials is controlled by the disengagement rate of the nucleobase chain ends from the "hard" phase, which, if shown to be general, provides a design criteria in the development of more thermally responsive materials. PMID- 16366574 TI - Quantum dot-based multiplexed fluorescence resonance energy transfer. AB - We demonstrate the use of luminescent quantum dots (QDs) conjugated to dye labeled protein acceptors for nonradiative energy transfer in a multiplexed format. Two configurations were explored: (1) a single color QD interacting with multiple distinct acceptors and (2) multiple donor populations interacting with one type of acceptor. In both cases, we showed that simultaneous energy transfer between donors and proximal acceptors can be measured. However, data analysis was simpler for the configuration where multiple QD donors are used in conjunction with one acceptor. Steady-state fluorescence results were corroborated by time resolved measurements where selective shortening of QD lifetime was measured only for populations that were selectively engaged in nonradiative energy transfer. PMID- 16366575 TI - Probing shapes of bichromophoric metal-organic complexes using ion mobility mass spectrometry. AB - Ion mobility mass spectrometry (IM-MS) was used to probe the structures of several metal complexes carrying pendant chromophores. The three complexes investigated were the copper(II) complex Cu(DAC)2+ (DAC = 1,8-bis(9 methylanthracyl)cyclam, cyclam = 1,4,8,11-tetraazacyclotetradecane), the N nitrosylated ligand DAC-NO, and the Roussin's red salt ester (mu-S,mu-S') protoporphyrin-IX-bis(2-thioethyl ester)tetranitrosyldiiron (PPIX-RSE). From the IM-MS data coupled with theoretical calculations, it was found that [Cu(II)(DAC - H)]+ exists as a single conformer, with one anthracenyl group above the cyclam and the other below, similar to the crystal structure of Cu(II)(DAC)2+. The metal free N-nitrosylated ligand (DAC-NO + H)+ has two conformations: one family of structures has one anthracenyl group above the cyclam and one below, while the other has both anthracenyl groups on the same side of the cyclam. These observations are consistent with 1H NMR data for the neutral DAC-NO complex that indicate the presence of two geometric isomers in solution. The third species, PPIX-RSE, has a porphyrin chromophore covalently linked to an Fe2S2(NO)4 cluster for use as a precursor for the photochemical delivery of nitric oxide in single- and two-photon excitation processes. Ion mobility indicates the presence of two (PPIX-RSE + H)+ conformations, consistent with the previous interpretation of the bimodal fluorescence lifetime decay seen for PPIX-RSE. DFT structures, in good agreement with the IM-MS cross sections, indicate two "bent" conformations with the planes of the porphyrin and Fe2S2 rings at different angles with respect to each other. PMID- 16366577 TI - Influence of thermal fluctuations on interfacial electron transfer in functionalized TiO2 semiconductors. AB - The influence of thermal fluctuations on the dynamics of interfacial electron transfer in sensitized TiO2-anatase semiconductors is investigated by combining ab initio DFT molecular dynamics simulations and quantum dynamics propagation of transient electronic excitations. It is shown that thermal nuclear fluctuations speed up the underlying interfacial electron transfer dynamics by introducing nonadiabatic transitions between electron acceptor states, localized in the vicinity of the photoexcited adsorbate, and delocalized states extended throughout the semiconductor material, creating additional relaxation pathways for carrier diffusion. Furthermore, it is shown that room-temperature thermal fluctuations reduce the anisotropic character of charge diffusion along different directions in the anatase crystal and make similar the rates for electron injection from adsorbate states of different character. The reported results are particularly relevant to the understanding of temperature effects on surface charge separation mechanisms in molecular-based photo-optic devices. PMID- 16366576 TI - Site-selective metal binding by designed alpha-helical peptides. AB - It is known that the designed alpha-helical peptide family TRI [(Ac-G(LKALEEK)4G CONH2)], containing single site substitution of a cysteine for a leucine, is capable of binding Cd(II) within a three-stranded coiled coil. The binding affinity of cadmium is dependent upon the site of substitution, with cysteine incorporated at the a site leading to cadmium complexes of higher affinity than when a d site was modified. In this work we have examined whether this differential binding affinity can be expressed in a di-cysteine-substituted peptide in order to develop site specificity within a designed system. The peptide TRI L9CL19C was used to determine whether significant differences in binding affinities at nearly proximal sites could be achieved in a short designed peptide. On the basis of 113Cd, 1H NMR, and circular dichroic spectroscopies, we have shown that 1 equiv of Cd(II) binds exclusively at the a site. Only after that position is filled does the second site become populated. Thus, the TRI system represents the first example where stoichiometrically equivalent peptides with different sequences form the framework for designing molecular assemblies that show site-specific ion recognition. We propose that the distinct metal affinities are due to the cysteine conformers at different substitution points along the peptide. Furthermore, we have shown that site selectivity in biomolecules can be encoded into relatively short peptides with helical sequences and, therefore, do not necessarily require the extensive protein scaffolds found in natural systems. PMID- 16366578 TI - Synthesis of stable dodecaalkoxy derivatives of hypercloso-B12H12. AB - The scope and limitations of the alkylation of [closo-B12(OH)12]2- using a series of fourteen alkyl and aralkyl halides and two p-toluenesulfonic acid esters in the presence of N,N-diisopropylethylamine have been investigated. The dodecaalkoxy-closo-dodecaborate products, [closo-B12(OR)12]2-, and their hypercloso two-electron oxidation products have been explored. The species [closo B12(OR)12]2- containing 26 cage-bonding electrons may undergo two reversible, sequential, one-electron oxidation processes, producing a 25-electron radical anion and a 24-electron neutral species. Several oxidizing reagents were investigated for the chemical oxidation of [closo-B12(OR)12]2- and [hypercloso B12(OR)12]- to [hypercloso-B12(OR)12]. Both FeCl3.6H2O and K3Fe(CN)6 in 90/5/5 ethanol/acetonitrile/H2O were found to be the reagents of choice. The reverse reaction leading from the neutral species to the radical anion and subsequently to the dianion was achieved using sodium borohydride in ethanol. A variety of alkoxyl derivatives have been synthesized by heating the reactants for extended periods of time in acetonitrile at the reflux temperature. The use of elevated reaction temperatures attained by employing moderate argon pressure (autoclave) over the reaction mixture led to drastic reductions in reaction times and increased efficiency. X-ray diffraction studies of substituted dodecabenzyl ether derivatives proved that 2(2-) has approximate Ih symmetry while hypercloso-2, -3, -9, -11, -12, and -13 have approximate D3d point group symmetry due to Jahn Teller distortion from Ih. PMID- 16366579 TI - Simulations of stochastic sensing of proteins. AB - We have performed Langevin dynamics and Poisson-Nernst-Planck calculations to simulate detection of proteins by genetically engineered alpha-hemolysin channels. In the recent stochastic sensing experiments, one end of a flexible polymer chain is permanently anchored inside the protein channel at a specified location, and the other end undergoes complexation with an analyte. Our simulations, using coarse-grained modeling, reproduce all essential qualitative results of the electrophysiology measurements of stochastic sensing. In addition, the underlying macromolecular mechanisms behind stochastic sensing are revealed in vivid details. The entropic fluctuations of the conformations of the tethered polymer chain dictate crucially the unique signatures of the ionic current trace of the channel and provide design rules for successful stochastic sensing. The origin of strong fluctuations in the ionic current of the channel is found to arise from the obstruction of the entrance at the beta-barrel of the channel by the fluctuating segments of the tether. Silencing of the pore is due to the suppression of conformational fluctuations of the chain, and the permanent blockade of ionic current is due to the threading of the tether through the channel. The onset of silencing and permanent blockade of the channel current cannot necessarily be attributed to the capture of analytes. In order for detection events to be timed accurately, the length and anchoring location of the tether must be tuned appropriately. PMID- 16366580 TI - Semiconductor rings fabricated by self-assembly of nanocrystals. AB - Three types of nonlinear organization schemes for creating ringlike semiconductors have been elucidated with the self-assembly of nanocrystals in aqueous phase under one-pot conditions. The synthetic architecture is based on morphological controls of primary nanocrystals which possess an intrinsic hexagonal symmetry. As most semiconducting materials have similar hexagonal (or cubic) crystal symmetries, in principle, the self-assembling processes revealed in this work should be extendable to other semiconductor nanocrystals. PMID- 16366581 TI - In situ spectroscopic study of the oxidation and reduction of Pd(111). AB - Using a photoemission spectroscometer that operates close to ambient conditions of pressure and temperature we have determined the Pd-O phase diagram and the kinetic parameters of phase transformations. We found that on the (111) surface oxidation proceeds by formation of stable and metastable structures. As the chemical potential of O2 increases chemisorbed oxygen forms followed by a thin surface oxide. Bulk oxidation is a two-step process that starts with the metastable growth of the surface oxide into the bulk, followed by a first-order transformation to PdO. PMID- 16366582 TI - Location, acid strength, and mobility of the acidic protons in Keggin 12 H3PW12O40: a combined solid-state NMR spectroscopy and DFT quantum chemical calculation study. AB - Solid-state 13C NMR experiments and quantum chemical Density Functional Theory (DFT) calculations of acetone adsorption were used to study the location of protons in anhydrous 12-tungstophosphoric acid (HPW), the mobility of the isolated and hydrated acidic protons, and the acid strength heterogeneity of the anhydrous hydroxyl groups. This study presents the first direct NMR experimental evidence that there are two types of isolated protons with different acid strengths in the anhydrous Keggin HPW. Rotational Echo DOuble Resonance (REDOR) NMR experiments combined with quantum chemical DFT calculations demonstrated that acidic protons in anhydrous HPW are localized on both bridging (Oc) and terminal (Od) atoms of the Keggin unit. The CP/MAS NMR experiments revealed that the isolated acidic protons are immobile, but hydrated acidic protons are highly mobile at room temperature. The isotropic chemical shift of the adsorbed acetone suggested that the acid strength of the H(H2O)n+ species in partially hydrated HPW is comparable to that of a zeolite, while the acidity of an isolated proton is much stronger than that of a zeolite. Isolated protons on the bridging oxygen atoms of anhydrous HPW are nearly superacidic. PMID- 16366583 TI - Cation dependence of chloride ion complexation by open-chained receptor molecules in chloroform solution. AB - Seventeen peptides, most having the sequence GGGPGGG, but differing in the C- and N-terminal ends, have been studied as anion-complexing agents. These relatively simple, open-chained peptide systems interact with both chloride and the associated cation. Changes in the N- and C-terminal side chains appear to make little difference in the efficacy of binding. NMR studies suggest that the primary interactions involve amide NH contacts with the chloride anion, and CD spectral analyses suggest a concomitant conformational change upon binding. Changes in binding constants, which are expected in different solvents, also suggest selective solvent interactions with the unbound host that helps to preorganize the open-chained peptide system. Significant differences are apparent in complexation strengths when the heptapeptide chain is shortened or lengthened or when the relative position of proline within the heptapeptide is varied. PMID- 16366584 TI - A general organocatalyst for direct alpha-functionalization of aldehydes: stereoselective C-C, C-N, C-F, C-Br, and C-S bond-forming reactions. Scope and mechanistic insights. AB - The development of a general organocatalyst for the alpha-functionalization of aldehydes, via an enamine intermediate, is presented. Based on optically active alpha,alpha-diarylprolinol silyl ethers, the scope and applications of this catalyst for the stereogenic formation of C-C, C-N, C-F, C-Br, and C-S bonds are outlined. The reactions all proceed in good to high yields and with excellent enantioselectivities. Furthermore, we will present mechanistic insight into the reaction course applying nonlinear effect studies, kinetic resolution, and computational investigations leading to an understanding of the properties of the alpha,alpha-diarylprolinol silyl ether catalysts. PMID- 16366585 TI - Raising the conductivity of crystalline polymer electrolytes by aliovalent doping. AB - Polymer electrolytes, salts dissolved in solid polymers, hold the key to realizing all solid-state devices such as rechargeable lithium batteries, electrochromic displays, or SMART windows. For 25 years conductivity was believed to be confined to amorphous polymer electrolytes, all crystalline polymer electrolytes were thought to be insulators. However, recent results have demonstrated conductivity in crystalline polymer electrolytes, although the levels at room temperature are too low for application. Here we show, for the first time, that it is possible to raise significantly the level of ionic conductivity by aliovalent doping. The conductivity may be raised by 1.5 orders of magnitude if the SbF6- ion in the crystalline conductor poly(ethylene oxide)6:LiSbF6 is replaced by less than 5 mol % SiF6(2-), thus introducing additional, mobile, Li+ ions into the structure to maintain electroneutrality. PMID- 16366587 TI - Controlling molecular crystal polymorphism with self-assembled monolayer templates. AB - The control of crystal polymorphism is a long-standing issue in solid-state chemistry, which has many practical implications for a variety of commercial applications. At least four different crystalline forms of 1,3-bis(m-nitrophenyl) urea (MNPU), a classic molecular crystal system, are known to crystallize from solution in various concomitant combinations. Herein we demonstrate that the introduction of gold-thiol self-assembled monolayers (SAMs) of substituted 4'-X mercaptobiphenyls (X = H, I, and Br) into the crystallization solution can serve as an effective means to selectively template the nucleation and growth of alpha , beta-, and gamma-MNPU phases, respectively. Polymorph control in the presence of SAM surfaces persists under a variety of solution conditions and consistently results in crystalline materials with high phase purity. The observed selectivity is rationalized on the basis of long-range two-dimensional geometric lattice matching and local complementary chemical interactions at the SAM/crystal interfaces. PMID- 16366586 TI - Structure and function of GDP-mannose-3',5'-epimerase: an enzyme which performs three chemical reactions at the same active site. AB - GDP-mannose-3',5'-epimerase (GME) from Arabidopsis thaliana catalyzes the epimerization of both the 3' and 5' positions of GDP-alpha-D-mannose to yield GDP beta-L-galactose. Production of the C5' epimer of GDP-alpha-D-mannose, GDP-beta-L gulose, has also been reported. The reaction occurs as part of vitamin C biosynthesis in plants. We have determined structures of complexes of GME with GDP-alpha-D-mannose, GDP-beta-L-galactose, and a mixture of GDP-beta-L-gulose with GDP-beta-L-4-keto-gulose to resolutions varying from 2.0 to 1.4 A. The enzyme has the classical extended short-chain dehydratase/reductase (SDR) fold. We have confirmed that GME establishes an equilibrium between two products, GDP beta-L-galactose and GDP-beta-L-gulose. The reaction proceeds by C4' oxidation of GDP-alpha-D-mannose followed by epimerization of the C5' position to give GDP beta-L-4-keto-gulose. This intermediate is either reduced to give GDP-beta-L gulose or the C3' position is epimerized to give GDP-beta-L-4-keto-galactose, then C4' is reduced to GDP-beta-L-galactose. The combination of oxidation, epimerization, and reduction in a single active site is unusual. Structural analysis coupled to site-directed mutagenesis suggests C145 and K217 as the acid/base pair responsible for both epimerizations. On the basis of the structure of the GDP-beta-L-gulose/GDP-beta-L-4-keto-gulose co-complex, we predict that a ring flip occurs during the first epimerization and that a boat intermediate is likely for the second epimerization. Comparison of GME with other SDR enzymes known to abstract a protein alpha to the keto function of a carbohydrate identifies key common features. PMID- 16366588 TI - Diazonium-protein adducts for graphite electrode microarrays modification: direct and addressed electrochemical immobilization. AB - Diazonium cation electrodeposition was investigated for the direct and electro addressed immobilization of proteins. For the first time, this reaction was triggered directly onto diazonium-modified proteins. Screen-printed (SP) graphite electrode microarrays were studied as active support for this immobilization. A 10-microelectrode (eight working electrodes, 0.2 mm2 each; one reference; and one auxiliary) setup was used to study the addressing possibilities of the method. These electrode microarrays were shown to be able to covalently graft diazonium cations through electrochemical reduction. Cyclic voltammetry and X-ray photoelectron spectroscopy were used to characterize the electrochemical grafting onto our SP graphite surface and suggested that a diazonium monolayer was deposited. Rabbit and human immunoglobulins (IgGs) were then chemically coupled to an aniline derivative (4-carboxymethylaniline), followed by diazotation to form an aryl diazonium function available for the electrodeposition. These modified proteins were both successfully electro-addressed at the surface of the graphite electrodes without cross-talk or interference. The immuno-biochip obtained using this novel approach enabled the specific detection of anti-rabbit IgG antibodies with a detection limit of 50 fmol of protein. A promising strategy to immobilize markedly different biological entities was then presented, providing an excellent spatial specificity of the electro-addressing. PMID- 16366590 TI - Semi-rational design of (north)-methanocarba nucleosides as dual acting A(1) and A(3) adenosine receptor agonists: novel prototypes for cardioprotection. AB - Ring-constrained adenosine analogues have been designed to act as dual agonists at tissue-protective A(1) and A(3) adenosine receptors (ARs). 9-Ribosides transformed into the ring-constrained (N)-methanocarba-2-chloro-5'-uronamides consistently lost affinity at A(1)/A(2A)ARs and gained at A(3)AR. Among 9 riboside derivatives, only N(6)-cyclopentyl and 7-norbornyl moieties were extrapolated for mixed A(1)/A(3) selectivity and rat/human A(3)AR equipotency. Consequently, 2 was balanced in affinity and potency at A(1)/A(3)ARs as envisioned and dramatically protected in an intact heart model of global ischemia and reperfusion. PMID- 16366591 TI - Human P2Y(6) receptor: molecular modeling leads to the rational design of a novel agonist based on a unique conformational preference. AB - Combining molecular dynamics (MD) in a hydrated phospholipid (DOPC) bilayer, a Monte Carlo search, and synthesis of locked nucleotide analogues, we discovered that the Southern conformation of the ribose is preferred for ligand recognition by the P2Y(6) receptor. 2'-Deoxy-(S)-methanocarbaUDP was found to be a full agonist of the receptor and displayed a 10-fold higher potency than that for the corresponding flexible 2'-deoxyUDP. MD results also suggested a conformational change of the second extracellular loop consequent to agonist binding. PMID- 16366592 TI - Conversion of the potent delta-opioid agonist H-Dmt-Tic-NH-CH(2)-bid into delta opioid antagonists by N(1)-benzimidazole alkylation(1). AB - N(1)-Alkylation of 1H-benzimidizole of the delta agonist H-Dmt-Tic-NH-CH(2)-Bid with hydrophobic, aromatic, olefinic, acid, ethyl ester, or amide (1-6) became delta antagonists (pA(2)=8.52-10.14). delta- and micro-Opioid receptor affinities were high (K(i)delta=0.12-0.36 nM and K(i)micro=0.44-1.42 nM). Only delta antagonism (pA(2)=8.52-10.14) was observed; micro agonism (IC(50)=30-450 nM) was not correlated with changes in alkylating agent or delta antagonism, and some compounds yielded mixed delta antagonism/micro agonism. PMID- 16366593 TI - Design, synthesis, and evaluation of pharmacological properties of cinnamic derivatives as antiatherogenic agents. AB - A series of cinnamic and phosphonocinnamic derivatives have been synthesized and their ability to inhibit cell-mediated LDL oxidation and oxidized LDL-induced cytotoxicity was investigated. Electron-donating substituents surrounding the necessary 4-OH group of the aromatic ring showed the best results. Among the different series tested, amide 1, thioester 5c, phosphonoester 7c, and the fluorophosphonocinnamic analogue 12c exhibited a potent inhibitory effect against LDL oxidation (and subsequent toxicity) mediated by cultured human microvascular endothelial cells (HMEC-1), with an efficacy comparable to that observed with probucol. Beside this indirect protective effect, these compounds exhibited a direct protective effect against the toxicity of previously oxidized LDL in HMEC 1. These data suggest that the newly synthesized cinnamic compounds should protect against early events (cell-mediated LDL oxidation) occurring within the vascular wall in atherosclerosis. PMID- 16366594 TI - Charge dependence of cellular uptake and selective antitumor activity of porphyrazines. AB - Porphyrazines (pzs), or tetraazaporphyrins, can be viewed as porphyrinic macrocycles in which the porphyrin meso (CH) groups are replaced by nitrogen atoms; as such, it can be anticipated that pzs would show similar biocompatibility and biodistribution to those of porphyrins. However, distinctive chemical and physical features of the pzs differentiate them from either the porphyrins or phthalocyanines, in particular making them excellent candidates as optical imaging/therapeutic agents. The novelty of the pzs requires that we first determine how specific structures selectively alter biological function, leading to the development of "rules" that will be used to predict future biologically functional pzs. In the first of these studies, we present here a correlation of pz charge with biocompatibility for a suite of three pzs-neutral, negative, and positive. Confocal fluorescence microscopy and proliferation/viability measurements disclose that the three pzs differ in their toxicity, uptake, and localization in A549 human lung adenocarcinoma cells and WI-38 VA13 normal cells. Interestingly, the negatively charged pz exhibits selective dark toxicity in pulmonary adenocarcinoma cells. PMID- 16366595 TI - Estradiol-adenosine hybrid compounds designed to inhibit type 1 17beta hydroxysteroid dehydrogenase. AB - The steroidogenic enzyme type 1 17beta-hydroxysteroid dehydrogenase (17beta-HSD) is involved in the synthesis of estradiol (E(2)), a hormone well-known to stimulate the growth of estrogen-sensitive tumors. To obtain compounds able to control E(2) formation, two moieties were linked with a methylene side chain: an adenosine moiety for interacting with the cofactor-binding site and an E(2) moiety for interacting with the substrate-binding site. When tested as inhibitors of type 1 17beta-HSD, the hybrid compounds inhibited the reductive activity (E(1) into E(2)) with IC(50) values ranging from 52 to 1,000 nM. The optimal side-chain length was determined to be eight methylene groups for a 16 beta-orientation. The presence of two components (E(2) and adenosine) is essential for good inhibition, since 16 beta-nonyl-E(2) and 5-nonanoyl-O-adenosine, two compounds having only one of the components, did not inhibit the enzyme. Moreover, the 3D-structure analysis of EM-1,745 complexed with type 1 17beta-HSD showed key interactions with both substrate- and cofactor-binding sites. PMID- 16366596 TI - Binding of rasagiline-related inhibitors to human monoamine oxidases: a kinetic and crystallographic analysis. AB - Monoamine oxidases A and B (MAO A and B) catalyze the degradation of neurotransmitters and represent drug targets for the treatment of neurodegenerative disorders. Rasagiline is an irreversible, MAO B-selective inhibitor that has been approved as a novel anti-Parkinson's drug. In this study, we investigate the inhibition of recombinant human MAO A and MAO B by several rasagiline analogues. Different substituents added onto the rasagiline scaffold alter the binding affinity depending on the position on the aminoindan ring and on the size of the substituent. Compounds with a hydroxyl group on either the C4 or the C6 atom inhibit both isozymes, whereas a bulkier substituent such as a carbamate is tolerated only at the C4 position. The 1.7 A crystal structure of MAO B in complex with 4-(N-methyl-N-ethyl-carbamoyloxy)-N-methyl-N-propargyl-1(R) aminoindan shows that the binding mode is similar to that of rasagiline with the carbamate moiety occupying the entrance cavity space. 1(R)-Aminoindan, the major metabolic product of rasagiline, and its analogues reversibly inhibit both MAO A and MAO B. The crystal structure of N-methyl-1(R)-aminoindan bound to MAO B shows that its aminoindan ring adopts a different orientation compared to that of rasagiline. PMID- 16366597 TI - Induced fit in mouse acetylcholinesterase upon binding a femtomolar inhibitor: a molecular dynamics study. AB - A molecular dynamics simulation of mouse acetylcholinesterase (mAChE) complexed with syn-TZ2PA6, a femtomolar AChE inhibitor, is compared to a simulation of unliganded mAChE. The simulation of the complex was initiated by placing the inhibitor in its bound conformation of the crystal complex into a structure of unliganded mAChE selected from preliminary protein-ligand docking results. During a 2 ns period, the enzyme subsequently displayed a substantial "induced fit" response to yield a conformation very similar to that obtained by crystallography (Bourne et al. Proc. Natl. Acad. Sci. U.S.A. 2004, 101, 1449-1454). In this conformation of unique nature, the Trp 286 side chain of the enzyme flips out of the hydrophobic core and becomes highly solvent exposed. The imidazole ring of His 287 is almost orthogonal relative to its position in the unliganded enzyme, creating a stable pi stacking arrangement with the Trp 286 side chain. Other major deviations among the active site residues include side chain conformational changes of Trp 86, Tyr 133, Tyr 337, and Phe 338. These residues in the complex deviate from their positions in unliganded mAChE to better accommodate the inhibitor in the active site gorge. PMID- 16366598 TI - (6,7-Dimethoxy-2,4-dihydroindeno[1,2-c]pyrazol-3-yl)phenylamines: platelet derived growth factor receptor tyrosine kinase inhibitors with broad antiproliferative activity against tumor cells. AB - A series of (6,7-dimethoxy-2,4-dihydroindeno[1,2-c]pyrazol-3-yl)phenylamines has been optimized to preserve both potent kinase inhibition activity against the angiogenesis target, the receptor tyrosine kinase of Platelet-Derived Growth Factor-BB (PDGF-BB), and to improve the broad tumor cell antiproliferative activity of these compounds. This series culminates in the discovery of 17 (JNJ 10198409), a compound with anti-PDGFR-beta kinase activity (IC(50)=0.0042 microM) and potent antiproliferative activity in six of eight human tumor cell lines (IC(50) < 0.033 microM). PMID- 16366599 TI - Kynurenamines as neural nitric oxide synthase inhibitors. AB - To find new compounds with potential neuroprotective activity, we have designed, synthesized, and characterized a series of neural nitric oxide synthase (nNOS) inhibitors with a kynurenamine structure. Among them, N-[3-(2-amino-5 methoxyphenyl)-3-oxopropyl]acetamide is the main melatonin metabolite in the brain and shows the highest activity in the series, with an inhibition percentage of 65% at a 1 mM concentration. The structure-activity relationship of the new series partially reflects that of the previously reported 2-acylamido-4-(2-amino 5-methoxyphenyl)-4-oxobutyric acids, endowed with a kynurenine-like structure. Structural comparisons between these new kinurenamine derivatives, kynurenines, and 1-acyl-3-(2-amino-5-methoxyphenyl)-4,5-dihydro-1H-pyrazole derivatives also reported confirm our previous model for the nNOS inhibition. PMID- 16366600 TI - N-substituted cis-4a-(3-hydroxyphenyl)-8a-methyloctahydroisoquinolines are opioid receptor pure antagonists. AB - N-Substituted cis-4a-(3-hydroxyphenyl)-8a-methyloctahydroisoquinolines (6a-g) were designed and synthesized as conformationally constrained analogues of the trans-3,4-dimethyl-4-(3-hydroxyphenyl)piperidine (4) class of opioid receptor pure antagonists. The methyloctahydroisoquinolines 6a-g can exist in conformations where the 3-hydroxyphenyl substituent is either axial or equatorial, similar to the (3-hydroxyphenyl)piperidines 4. The 3-hydroxyphenyl equatorial conformation is responsible for the antagonist activity observed in the (3-hydroxyphenyl)piperidine antagonists. Single-crystal X-ray analysis of 6a shows that the 3-hydroxyphenyl equatorial conformation is favored in the solid state. Molecular modeling studies also suggest that the equatorial conformation has lower potential energy relative to that of the axial conformation. Evaluation of 6a-g in the [(35)S]GTP-gamma-S in vitro functional assay showed that they were opioid receptor pure antagonists. N-[4a-(3-Hydroxyphenyl)-8a-methyl-2-(3 phenylpropyl)octahydroisoquinoline-6-yl]-3-(piperidin-1-yl)propionamide (6d) with a K(e) of 0.27 nM at the kappa opioid receptor with 154- and 46-fold selectivity relative to those of the micro and delta receptors, respectively, possessed the best combination of kappa potency and selectivity. PMID- 16366601 TI - Novel indole-based peroxisome proliferator-activated receptor agonists: design, SAR, structural biology, and biological activities. AB - The synthesis and structure-activity relationship studies of novel indole derivatives as peroxisome proliferator-activated receptor (PPAR) agonists are reported. Indole, a drug-like scaffold, was studied as a core skeleton for the acidic head part of PPAR agonists. The structural features (acidic head, substitution on indole, and linker) were optimized first, by keeping benzisoxazole as the tail part, based on binding and functional activity at PPARgamma protein. The variations in the tail part, by introducing various heteroaromatic ring systems, were then studied. In vitro evaluation led to identification of a novel series of indole compounds with a benzisoxazole tail as potent PPAR agonists with the lead compound 14 (BPR1H036) displaying an excellent pharmacokinetic profile in BALB/c mice and an efficacious glucose lowering activity in KKA(y) mice. Structural biology studies of 14 showed that the indole ring contributes strong hydrophobic interactions with PPARgamma and could be an important moiety for the binding to the protein. PMID- 16366602 TI - A new bisintercalating anthracycline with picomolar DNA binding affinity. AB - A new bisintercalating anthracycline (WP762) has been designed, in which monomeric units of daunorubicin have been linked through their amino groups on the daunosamine moieties using an m-xylenyl linker. Differential scanning calorimetry and UV melting experiments were used to measure the ultratight binding of WP762 to DNA. The binding constant for the interaction of WP762 with herring sperm DNA was determined to be 7.3 (+/-0.2) x 10(12) M(-1) at 20 degrees C. The large favorable binding free energy of -17.3 kcal mol(-1) was found to result from a large negative enthalpic contribution of -33.8 kcal mol(-1) and an opposing entropic term (-TDeltaS = +16.5 kcal mol(-1)). A comparative molecular modeling study rationalized the increased binding by the m-xylenyl linker of WP762 positioning in the DNA minor groove compared to the p-xylenyl linker found in WP631, the first bis-anthracycline of this type. The cytotoxicity of WP762 was compared to that of other anthracyclines in Jurkat T lymphocytes. These studies, together with an analysis of the cell-cycle traverse in the presence of WP762, suggest that in these cells the new drug is more cytotoxic than the structurally related WP631. PMID- 16366603 TI - Comparison of the NO and HNO donating properties of diazeniumdiolates: primary amine adducts release HNO in Vivo. AB - Diazeniumdiolates, more commonly referred to as NONOates, have been extremely useful in the investigation of the biological effects of nitric oxide (NO) and related nitrogen oxides. The NONOate Angeli's salt (Na(2)N(2)O(3)) releases nitroxyl (HNO) under physiological conditions and exhibits unique cardiovascular features (i.e., positive inotropy/lusitropy) that may have relevance for pharmacological treatment of heart failure. In the search for new, organic-based compounds that release HNO, we examined isopropylamine NONOate (IPA/NO; Na[(CH(3))(2)CHNH(N(O)NO]), which is an adduct of NO and a primary amine. The chemical and pharmacological properties of IPA/NO were compared to those of Angeli's salt and a NO-producing NONOate, DEA/NO (Na[Et(2)NN(O)NO]), which is a secondary amine adduct. Under physiological conditions IPA/NO exhibited all the markers of HNO production (e.g., reductive nitrosylation, thiol reactivity, positive inotropy). These data suggest that primary amine NONOates may be useful as HNO donors in complement to the existing series of secondary amine NONOates, which are well-characterized NO donors. PMID- 16366604 TI - Synthesis, biological activity, and preliminary pharmacokinetic evaluation of analogues of a phosphosulfomannan angiogenesis inhibitor (PI-88). AB - The phosphosulfomannan 1 (PI-88) is a mixture of highly sulfated oligosaccharides that is currently undergoing clinical evaluation in cancer patients. As well as its anticancer properties, 1 displays a number of other interesting biological activities. A series of analogues of 1 were synthesized with a single carbon (pentasaccharide) backbone to facilitate structural characterization and interpretation of biological results. In a fashion similar to 1, all compounds were able to inhibit heparanase and to bind tightly to the proangiogenic growth factors FGF-1, FGF-2, and VEGF. The compounds also inhibited the infection of cells and cell-to-cell spread of herpes simplex virus (HSV-1). Preliminary pharmacokinetic data indicated that the compounds displayed different pharmacokinetic behavior compared with 1. Of particular note was the n-octyl derivative, which was cleared 3 times less rapidly than 1 and may provide increased systemic exposure. PMID- 16366605 TI - Methyl substitution on the piperidine ring of N-[omega-(6-methoxynaphthalen-1 yl)alkyl] derivatives as a probe for selective binding and activity at the sigma(1) receptor. AB - The N-(6-methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)propyl and N-(6 methoxynaphthalen-1-yl)propyl derivatives as well as their upper homologous butyl derivatives of various methylpiperidines were prepared. The piperidine moiety bearing monomethyl or geminal dimethyl groups was employed as a probe to explore sigma-subtype affinities and selectivities by radioligand binding assays at sigma(1) and sigma(2) receptors and the Delta(8)-Delta(7) sterol isomerase (SI) site. 4-Methyl derivative 31 was the most potent sigma(1) ligand (K(i)=0.030 nM) with a good selectivity profile (597-fold and 268-fold relative to sigma(2) receptor and SI site, respectively), whereas 3,3-dimethyl derivative 26 (K(i)=0.35 nM) was the most selective (680-fold) relative to the sigma(2) receptor. Both compounds can be proposed as tools for PET experiments. Furthermore, the naphthalene compounds 26, 28, 31, and 33 demonstrated antiproliferative activity in rat C6 glioma cells (EC(50) = 15.0 microM for 33), revealing a putative sigma(1) antagonist activity and opening a useful perspective in tumor research and therapy. PMID- 16366606 TI - New potential anticancer agents based on the anthranilic acid scaffold: synthesis and evaluation of biological activity. AB - The synthesis and anticancer activity of new compounds designed on the anthranilic acid scaffold are reported. The antiproliferative activity was assayed by the National Cancer Institute in established in vitro and in vivo anticancer experimental models. Structural variations based on the flufenamic acid motif afforded a series of (hetero)aryl esters of N-(2 (trifluoromethyl)pyridin-4-yl)anthranilic acid, which showed in vitro growth inhibitory properties against human tumor cell lines in nanomolar to low micromolar concentrations. The pyridinyl ester 25 exhibited very potent in vitro antiproliferative efficacy, with a chemosensitive profile showing a number of GI(50) values at concentrations lower than 10(-7) M in the full panel of human tumor cell lines. Compound 25 was also tested in vivo as a potential anticancer agent in the hollow fiber assay and in human tumor xenografts, showing moderate inhibitory properties. Analysis of biological activities and the COMPARE procedure was utilized to support putative biochemical mechanisms implicated with the antiproliferative activity. PMID- 16366607 TI - 2-(Benzimidazol-2-yl)quinoxalines: a novel class of selective antagonists at human A(1) and A(3) adenosine receptors designed by 3D database searching. AB - The Cambridge Structural Database (CSD) was searched through two 3D queries based on substructures shared by well-known antagonists at the A(1) and A(3) adenosine receptors (ARs). Among the resulting 557 hits found in the CSD, we selected five compounds to purchase, synthesize, or translate synthetically into analogues better tailored to interact with the biological targets. Binding experiments using human ARs showed that four out of five tested compounds turned out to be antagonists at the A(1)AR or A(3)AR with K(i) values between 50 and 440 nM. Lead optimizations of 2-(benzimidazol-2-yl)quinoxalines (BIQs, 3) gave the best results in terms of potency and selectivity at the A(1) and A(3) ARs. Particularly, 2-(4-ethylthiobenzimidazol-2-yl)quinoxaline (3e) exhibited K(i) values at the A(1)AR, A(2A)AR, and A(3)AR of 0.5, 3440, and 955 nM, respectively, whereas 2-(4-methylbenzimidazol-2-yl)quinoxaline (3b) displayed at the same ARs K(i) values of 8000, 833, and 26 nM, respectively. PMID- 16366608 TI - Synthesis and evaluation of cyclic secondary amine substituted phenyl and benzyl nitrofuranyl amides as novel antituberculosis agents. AB - In an ongoing effort to develop new and potent antituberculosis agents, a second generation series of nitrofuranyl amides was synthesized on the basis of the lead compound 5-nitrofuran-2-carboxylic acid 3,4-dimethoxybenzylamide. The primary design consideration was to improve the solubility and consequently the bioavailability of the series by the addition of hydrophilic rings to the benzyl and phenyl B ring core. The synthesis of 27 cyclic, secondary amine substituted phenyl and benzyl nitrofuranyl amides is described and their activity against Mycobacterium tuberculosis reported. The series showed a strong structure activity relationship as the benzyl nitrofuranyl amides were significantly more active than similarly substituted phenyl nitrofuranyl amides. Para-substituted benzyl piperazines showed the most antituberculosis activity. Compounds in the series were subsequently selected for bioavailability and in vivo testing. This study led to the successful discovery of novel compounds with increased antituberculosis activity in vitro and a better understanding of the requisite pharmacological properties to advance this class. PMID- 16366609 TI - Synthesis of tricyclic 1,3-oxazin-4-ones and kinetic analysis of cholesterol esterase and acetylcholinesterase inhibition. AB - A series of thieno[1,3]oxazin-4-ones and thieno[1,3]thiazin-4-ones were synthesized and investigated as inhibitors of the alpha/beta hydrolases cholesterol esterase (CEase) and acetylcholinesterase (AChE). The introduction of a cycloaliphatic five- or six-membered ring fused at the thiophene was favorable for CEase inhibition. Such compounds were analyzed as true alternate substrate inhibitors. 6,7-Dihydro-2-(dimethylamino)-4H,5H-cyclopenta[4,5]thieno[2,3 d][1,3]oxazin-4-one (33) exhibited a K(i) value of 630 nM and excelled in its low susceptibility to CEase-catalyzed degradation. Compound 33 and its analogues did not inhibit AChE. The introduction of a tetrahydropyrido ring with bulky hydrophobic substituents at the basic nitrogen provided inhibitors of AChE which were completely inactive toward CEase. 7-Benzyl-5,6,7,8-tetrahydro-2-(N-3,4 dimethoxybenzyl-N-methylamino)-4H-pyrido[4',3':4,5]thieno[2,3-d][1,3]oxazin-4-one (21) had the IC(50) value of 330 nM for AChE inhibition. A residual enzymatic activity at an infinite inhibitor concentration and thus a catalytically active ternary enzyme-substrate-inhibitor complex was concluded. To specify kinetic parameters of inhibition, a new method was derived to characterize selected thieno[1,3]oxazin-4-ones as hyperbolic mixed-type inhibitors of AChE. PMID- 16366611 TI - Can a single-reference approach provide a balanced description of ground and excited states? A comparison of the completely renormalized equation-of-motion coupled-cluster method with multireference quasidegenerate perturbation theory near a conical intersection and along a photodissociation coordinate in ammonia. AB - We calculated the two lowest electronically adiabatic potential energy surfaces of ammonia in the region of the conical intersection and at a sequence of geometries along which one of the N-H bonds is broken. We employed both a multireference (MR) method and a single-reference (SR) method. The MR calculations are based on multiconfiguration quasidegenerate perturbation theory (MC-QDPT) with a 6-311+G(3df,3pd) basis set. The SR calculations, carried out with the same basis, employ the completely renormalized equation-of-motion coupled-cluster method with singles and doubles, and a noniterative treatment of triples, denoted CR-EOMCCSD(T). At 91 geometries used for comparison, including geometries near a conical intersection, the surfaces agree to 7% on average. PMID- 16366612 TI - Aromaticity in stable tiara nickel thiolates: computational and structural analysis. AB - Quantum chemical calculations as well as crystallographic analyses show that the Ni(n) rings in the tiara Ni thiolates, [NiS2]n and [Ni(SR)2]n (n = 3-6), have highly symmetric polygonal structures. We find that such structural features primarily arise from the effective delocalization of the d-orbital electrons across the Ni(n) rings leading to bond length equalization and thereby aromaticity. We introduce the d-orbital aromaticity for the first time to explain the experimentally observed polygonal structures of these cyclic metal rings bridged by thiol linkages. PMID- 16366610 TI - Preparation and biological evaluation of indole, benzimidazole, and thienopyrrole piperazine carboxamides: potent human histamine h(4) antagonists. AB - Three series of H(4) receptor ligands, derived from indoly-2-yl-(4-methyl piperazin-1-yl)-methanones, have been synthesized and their structure-activity relationships evaluated for activity at the H(4) receptor in competitive binding and functional assays. In all cases, substitution of small lipophilic groups in the 4 and 5-positions led to increased activity in a [(3)H]histamine radiolabeled ligand competitive binding assay. In vitro metabolism and initial pharmacokinetic studies were performed on selected compounds leading to the identification of indole 8 and benzimidazole 40 as potent H(4) antagonists with the potential for further development. In addition, both 8 and 40 demonstrated efficacy in in vitro mast cell and eosinophil chemotaxis assays. PMID- 16366613 TI - Syntheses, structures, and photoisomerization of (E)- and (Z)-2-tert-butyl-9 (2,2,2-triphenylethylidene)fluorene. AB - "Sterically geared" 9-(2,2,2-triphenylethylidene)fluorene (1) is of potential interest as a photoactive moiety in molecular devices, and the 2-tert-butyl derivative (6) has been synthesized to investigate photoisomerization. E and Z stereoisomers of 6 were separated and identified by X-ray crystallography. The tert-butyl group does not introduce additional strain, and its close proximity to the trityl group in the Z isomer suggests an attractive van der Waals interaction. The UV spectra of (E)-6 and (Z)-6 are nearly identical, showing absorption bands that are similar to those of fluorene occurring at wavelengths longer than 240 nm. Photoisomerization of 6 was investigated at 266, 280 and 320 nm. Solutions initially containing only (E)-6 or (Z)-6 were irradiated with pulsed laser light, monitoring isomerization by 1H NMR spectroscopy. Negligible photodecomposition was observed when the solutions were agitated by N2 ebullition. Experimental data were fitted to theoretical curves to obtain photoisomerization quantum yields (phi(ZE) and phi(EZ)) ranging from 0.04 to 0.09. This first photoisomerization study of a dibenzofulvene reveals significant quantum yields, despite theoretical prediction of inefficient or negligible isomerization of the parent hydrocarbon, fulvene. Thermal isomerization of 6 at 270 degrees C (t(1/2) = 120 min) was also followed by 1H NMR spectroscopy, resulting in an estimated activation energy (deltaG(double dagger)) of 43 kcal/mol. PMID- 16366615 TI - Time-resolved EPR study of the photophysics and photochemistry of 1-(3 (methoxycarbonyl)propyl)-1-phenyl[6.6]C61. AB - Time-resolved (TR) EPR was used to study the photophysics and photochemistry of 1 (3-(methoxycarbonyl)propyl)-1-phenyl[6.6]C61 (M1). The CW TREPR spectra of M1 in the photoexcited triplet state, frozen in a rigid matrix and in liquid solution at room temperature, were compared with those of 3C60. The introduction of the substituent on C60 has a striking effect on the spectra of the triplets, which is attributed to the lifting of the orbital degeneracy by the reduction in symmetry. Fourier transform (FT) EPR was used in an investigation of electron-transfer reactions in liquid solutions mediated by 3M1. Of particular interest was the system of M1/chloranil (CA)/perylene (Pe). Photoexcitation of M1 is found to lead to the formation of the chloranil anion radical and the perylene cation radical. From the chemically induced dynamic electron polarization (CIDEP) patterns in the FTEPR spectra and the dependence of the reaction kinetics on reactant concentrations, it was deduced that CA- is formed by two competing pathways following photoexcitation of M1: (1) direct electron transfer from 3M1 to CA followed by electron transfer from Pe to M1+ and (2) energy transfer from 3M1 to Pe followed by oxidative quenching of 3Pe by CA. In both pathways, M1 acts as a light-energy harvester and mediator of electron-transfer reactions from Pe to CA without itself being consumed in the process, that is, as a photocatalyst. It is found that the functionalization of C60 makes its triplet state a worse electron donor and acceptor, but it has no significant effect on the triplet energy transfer reaction. PMID- 16366614 TI - Identifiability of models for intramolecular two-state excited-state processes with added quencher and coupled species-dependent rotational diffusion. AB - The parameters describing the kinetics of excited-state processes can possibly be recovered by analysis of the fluorescence decay surface measured as a function of the experimental variables. The identifiability analysis of a photophysical model assuming errorless time-resolved fluorescence data can verify whether the model parameters can be determined and may lead to the minimal experimental conditions under which this is possible. In this work, we used the method of similarity transformation to investigate the identifiability of three kinetic models utilized to describe the time-resolved fluorescence of reversible intramolecular two-state excited-state processes in isotropic environments: (1) model without added quencher, (2) model with added quencher, (3) model with added quencher coupled with species-dependent rotational diffusion described by Brownian reorientation. Without a priori information, model 1 is not identifiable. For model 2, two sets of quenching rate constants and combinations of excited-state deactivation/exchange rate constants are possible, but they cannot be allocated to a specific excited-state species. For both sets, upper and lower limits on the excited-state deactivation/exchange rate constants can be obtained. For model 3, both spherically and cylindrically symmetric rotors, with no change in the principal axes of rotation in the latter, are considered. The fluorescence delta response functions I(parallel)(t) and I(perpendicular)(t), for fluorescence polarized parallel and perpendicular, respectively, to the electric vector of linearly polarized excitation, are used to define the sum S(t) identically equal to I(parallel)(t) + 2 I(perpendicular)(t) and the difference D(t) identically equal to I(parallel)(t) - I(perpendicular)(t). The identifiability analysis is performed using the S(t) and D(t) functions. Also for model 3, two sets of kinetic parameters (i.e., quenching rate constants, combinations of deactivation/exchange rate constants, and rotational diffusion coefficients) exist, but these parameters cannot be assigned unequivocally to a specific species. For the three models, an infinite number of alternative spectroscopic parameters associated with excitation and emission are found. PMID- 16366616 TI - Vibrational energy flow rates for cis- and trans-stilbene isomers in solution. AB - Transient electronic absorption following excitation of the first C-H stretching overtone (2nu(CH)) or a C-H stretch-bend combination (nu(CH) + nu(bend)) monitors the flow of vibrational energy in cis-stilbene and in trans-stilbene. Following a rapid initial rise as energy flows into states interrogated by the probe pulse, the absorption decays with two time constants, which are about a factor of 2 longer for the cis-isomer than for the trans-isomer. The decay times for cis stilbene are tau2(cis) = (2.6 +/- 1.5) ps and tau3(cis) = (24.1 +/- 2.1) ps, and those for trans-stilbene are tau2(trans) = (1.4 +/- 0.6) ps and tau3(trans) = (10.2 +/- 1.1) ps. The decay times are essentially the same in different solvents, suggesting that the relaxation is primarily intramolecular. The two decay times are consistent with the sequential flow of energy through sets of coupled states within the molecule, and the difference in the rates for the two isomers likely reflects differences in coupling among the states arising from the different structures of the isomers. The similarity of the time evolution following excitation of the first C-H overtone at 5990 cm(-1) and the stretch bend combination at 4650 cm(-1) is consistent with a subset of states, whose structure is similar for the two vibrational excitation energies, controlling the observed flow of energy. PMID- 16366617 TI - Free electron transfer from xanthenyl- and fluorenylsilanes (Me3 or Ph3) to parent solvent radical cations: effects of molecule dynamics. AB - Parent radical cations of nonpolar solvents (alkanes and alkyl chlorides) ionize 9-(trimethylsilyl)xanthenes and 9-(trimethylsilyl)fluorenes in a diffusion controlled electron transfer. The actual electron jump as the deciding part of the process does not require a defined encounter complex, and therefore the reactants are not subjected to any geometry optimization. Considering the molecule dynamics of the donors, bending motions of the silyl group are concerted with fluctuations of the highest occupied molecular orbital electrons. Ionizing such a standing conformer mixture creates metastable (microsecond) as well as dissociative donor radical cations. A mobility restriction of the benzylic silane group in positions vertical to the phenyl plane stabilizes the radical cations and accounts for a declining amount of dissociative radical cations, which undergo C-Si bond fragmentation in the order benzylsilane > xanthenylsilane > fluorenylsilane. PMID- 16366618 TI - Charge transfer processes in conjugated triarylamine-oligothiophene perylenemonoimide dendrimers. AB - The synthesis and charge transfer properties of triarylamine-oligothiophene perylenemonoimide dendrimers, TPA(T2-PMI)3 and TPA(T4-PMI)3, are described. The fluorescence quantum yields indicate strong emission quenching by electron transfer [phi(THF) = 0.004 for TPA(T2-PMI)3, phi(THF) = 0.003 for TPA(T4-PMI)3, and phi(THF) = 0.8 for PMI]. Moreover, with the increase of the solvent polarity, the quantum yields decrease indicating that the A+* D-* (acceptor/donor) couple is more stabilized. The femtosecond transient absorption spectra show a very fast charge separation process (approximately 2 ps; k(cs) approximately 5 x 10(11) s( 1)) and a charge recombination of more than 1 order of magnitude slower (approximately 50 ps; k(cr) approximately 2 x 10(10) s(-1)), as observed from the rise time and decay of the radical anion and radical cation absorption bands. The analysis of the transient absorption spectroscopy and of the energetics of the process using Marcus theory indicates that in the electron transfer process the thiophene unit is the first electron donor. The triarylamine is not functioning as a second electron donor, as also substantiated by the absence of an effect of the addition of acid on the emission intensity of the dendrimers. The presence of the triarylamine and/or the proximity of the oligothiophenes does improve the donor capabilities of the oligothiophene unit slightly and enhances its conjugation as seen in the absorption spectra and the transients of the radial cations. These results can be used for a better design of molecular materials for, e.g., photovoltaic applications. PMID- 16366619 TI - Spectroscopy and femtosecond dynamics of 7-N,N-diethylamino-3-hydroxyflavone. The correlation of dipole moments among various states to rationalize the excited state proton transfer reaction. AB - Comprehensive excitation behaviors of 7-N,N-diethylamino-3-hydroxyflavone (I) have been investigated via steady state, temperature-dependent emission, and fluorescence upconversion to probe the excited-state intramolecular proton transfer (PT) reaction. Upon excitation, I undergoes ultrafast (<<120 fs), adiabatic type of charge transfer (CT), so that the dipolar vector in the Franck Condon excited state is much different from that in the ground state. In polar solvents such as CH2Cl2 and CH3CN, early relaxation dynamics clearly reveals the competitive rates between solvent relaxation and PT dynamics. After reaching thermal equilibrium, a relatively slow, solvent-polarity-dependent rate (a few tens of picoseconds(-1)) of PT takes places. Firm support of the early relaxation dynamics is rendered by the spectral temporal evolution, which resolves two distinct bands ascribed to CT and PT emission. The results, in combination with ab initio calculations on the dipolar vectors for various corresponding states, led us to conclude that excited-state normal (N*) and excited proton-transfer tautomer (T*) possesses very different dipole orientation, whereas the dipole orientation of the normal ground state (N) is between that of N* and T*. PT is thus energetically favorable at the Franck-Condon excited N*, and its rate is competitive with respect to the solvent relaxation dynamics induced by CT. Unlike the well-known PT system, 4'-N,N-diethylamino-3-hydroxyflavone, in which equilibrium exists between solvent-equilibrated N(eq)* and T(eq)*, N(eq)* --> T(eq)* PT for I is a highly exergonic, irreversible process in all solvents studied. Further temperature-dependent studies deduce a solvent-polarity perturbed energy barrier of 3.6 kcal/mol for the N(eq)* --> T(eq)* PT in CH3CN. The proposed dipole-moment-tuning PT mechanism with the associated relaxation dynamics is believed to apply to many PT molecules in polar, aprotic solvents. PMID- 16366620 TI - Ground- and excited-state double proton transfer in lumichrome/acetic acid system: theoretical and experimental approach. AB - Experimental time-resolved spectral and photon counting kinetic results confirm formation of an isoalloxazinic excited state via excited-state double proton transfer (ESDPT) catalyzed by a carboxylic acid molecule that forms a hydrogen bond complex with the parent alloxazine molecule. This isoalloxazinic tautomer manifests itself as a distinct long-lived emissive species formed only in such alloxazine derivatives that were not substituted at the N1 nitrogen atom, being a product of the excited-state reaction occurring from the alloxazinic excited state. Theoretical calculations support the idea that the ESDPT occurs by the concerted mechanism. The calculated activation barrier in the excited state is much lower than the same barrier in the ground state and even disappears for the HOMO-1 to LUMO excitation, which explains the fact that the reaction takes place in the excited-state only. The reaction rate estimated from the emission kinetics is ca. 1.4 x 10(8) dm3 mol(-1) s(-1) in ethanolic solutions of lumichrome with added acetic acid. PMID- 16366621 TI - Intra- vs intermolecular photoinduced electron transfer reactions of a macrocyclic donor-acceptor dyad. AB - The synthesis, structural characterization, and photophysical behavior of a 14 membered tetraazamacrocycle with pendant 4-dimethylaminobenzyl (DMAB) and 9 anthracenylmethyl groups is reported (L3, 6-((9-anthracenylmethyl)amino)-trans 6,13-dimethyl-13-((4-dimethylaminobenzyl)amino)-1,4,8,11 tetraazacyclotetradecane). In its free base form, this compound displays rapid intramolecular photoinduced electron transfer (PET) quenching of the anthracene emission, with both the secondary amines and the DMAB group capable of acting as electron donors. When complexed with Zn(II), the characteristic fluorescence of the anthracene chromophore is restored as the former of these pathways is deactivated by coordination. Importantly, it is shown that the DMAB group, which remains uncoordinated and PET active, acts only very weakly to quench emission, by comparison to the behavior of a model Zn complex lacking the pendant DMAB group, [ZnL2]2+ (Chart 1). By contrast, Stern-Volmer analysis of intermolecular quenching of [ZnL2]2+ by N,N-dimethylaniline (DMA) has shown that this reaction is diffusion limited. Hence, the pivotal role of the bridge in influencing intramolecular PET is highlighted. PMID- 16366622 TI - Linear dichroism in the X-ray absorption spectra of linear n-alkanes. AB - The nature of the linear dichroism in the near-edge X-ray absorption fine structure (NEXAFS) spectra of linear n-alkanes is a matter of long-standing controversy. Linear dichroism in the carbon 1s --> sigma*(C-C) transition has been interpreted within a building block model and a molecular orbital model, leading to two different descriptions for the angular dependence of this feature. When used for measurement of molecular orientation, the application of these two different models will lead to different results. We have explored the linear dichroism in the carbon 1s NEXAFS spectra of single crystals of the linear n alkane hexacontane (n-C60H122). An analysis of the angular dependence in this spectrum shows that the transition dipole moment associated with the carbon 1s - > sigma*(C-C) transition is oriented along the macromolecular chain axis, contradicting the predictions of the building block model. However, other transitions are observed in the sigma*(C-H) and the sigma*(C-C) bands that are orthogonal to the dominant transitions for each band. We also observe that radiation damage can be manifest in the form of molecular reorientation in highly ordered organic thin films. PMID- 16366623 TI - Reaction path of UV photolysis of matrix isolated acetyl cyanide: formation and identification of ketenes, zwitterion, and keteneimine intermediates. AB - Irradiations at lambda > 180 nm and lambda > 230 nm of CH3COCN (CD3COCN) trapped in an argon matrix at 10 K have been performed and monitored by Fourier transform infrared spectroscopy. For both wavelengths, the first photoreaction product is acetyl isocyanide. At lambda > 180 nm acetonitrile and methyl isocyanide are obtained as final products. They are formed by photolysis of acetyl isocyanide and acetyl cyanide, respectively. Unstable intermediates, such as ketene:HCN and ketene:HNC complexes, H2CNCH zwitterion, and H2C2NH keteneimine, not observed in the gas phase, are trapped and identified at our experimental temperature. The complexes have an L-shaped structure with a hydrogen bond between the oxygen atom of ketene and the hydrogen atom of HCN or HNC. A pathway process is proposed and compared with the ones determined in the ground state by ab initio calculations. PMID- 16366624 TI - Phenyl-coinage metal (Ag, Au) complexes: an anion photoelectron spectroscopy and density functional study. AB - The important intermediate phenyl-coinage metal complexes (Ag(m)C6H5(-), Au(m)C6H5(-)), which are produced from the reactions between coinage metal clusters formed by laser ablation and the benzene molecules seeded in argon carrier gas, are studied by PES (photoelectron spectroscopy) and DFT (density functional theory). The EAs (adiabatic electron affinities) of these complexes are obtained from PES at both 308 and 193 nm photon energies and show odd-even alternation. Calculations with DFT are carried out on the structural and electronic properties of Ag(m)C6H5(-) and Au(m)C6H5(-); the adiabatic detachment energy and the calculated DOS (density of states) for the ground state of a given anion are in good agreement with the experimental PES results. The observed spectra are also compared with those of the pure coinage metal clusters, which reveal that there are some similarities between them and the phenyl acts like an additional metal atom in the clusters. Furthermore, the bonding between phenyl and metal is analyzed, suggesting that phenyl group binds perpendicularly on metal clusters through C-M sigma bond. PMID- 16366625 TI - Nonresonant optical activity of isolated organic molecules. AB - Cavity ring-down polarimetry (CRDP) has been exploited to interrogate the nonresonant optical activity (or circular birefringence) of prototypical organic compounds in the vapor phase, thereby revealing the intrinsic chiro-optical response evoked from isolated (solvent-free) molecules. Specific polarization rotation parameters have been measured at two distinct excitation wavelengths (355 nm and 633 nm) for a variety of gas-phase species drawn from the terpene, epoxide, and alkane/alkene families, with complementary solution-phase polarimetric studies serving to highlight the pronounced influence of solute solvent interactions. Time-dependent linear response calculations performed at high levels of density functional theory have been enlisted to unravel the structural and electronic origins for observed behavior. Aside from elucidating the complex solvation processes that mediate chiro-optical phenomena taking place in condensed media, this study affords a critical assessment for emerging ab initio predictions of nonresonant optical activity and for their promising ability to assist in the determination of absolute molecular stereochemistry. PMID- 16366626 TI - Coordination of ScO+ and YO+ by multiple Ar, Kr, and Xe atoms in noble gas matrixes: a matrix isolation infrared spectroscopic and theoretical study. AB - The combination of matrix isolation infrared spectroscopic and quantum chemical calculation results provide strong evidence that scandium and yttrium monoxide cations, ScO+ and YO+, coordinate multiple noble gas atoms in forming noble gas complexes. The results showed that ScO+ coordinates five Ar, Kr, or Xe atoms, and YO+ coordinates six Ar or Kr and five Xe atoms in solid noble gas matrixes. Hence, the ScO+ and YO+ cations trapped in solid noble gas matrixes should be regarded as the [ScO(Ng)5]+ (Ng = Ar, Kr, or Xe), [YO(Ng)6]+ (Ng = Ar or Kr) or [YO(Xe)5]+ complexes. Experiments with dilute krypton or xenon in argon or krypton in xenon produced new IR bands, which are due to the stepwise formation of the [ScO(Ar)(5-n)(Kr)n]+, [ScO(Kr)(5-n)(Xe)n]+ (n = 1-5), [YO(Ar)(6-n)(Kr)n]+ (n = 1-6), and [YO(Ar)(6-n)(Xe)n]+ (n = 1-4) complexes. PMID- 16366627 TI - Electronic structure of the hydroxo and methoxo oxometalate anions MO3(OH)- and MO3(OCH3)- (M = Cr, Mo, and W). AB - The electronic structure of the mononuclear hydroxo MO3(OH)- and methoxo MO3(OCH3)- Group 6 oxometalate anions (M = Cr, Mo, and W) were examined by photodetachment photoelectron spectroscopy and electronic structure calculations at the density functional and CCSD(T) levels of theory. All of the anions exhibited high electron binding energies (>4.9 eV), with the lowest-energy detachment features arising from oxygen 2p-based orbitals. The combined experimental and theoretical results allowed the change in molecular orbital energy levels to be investigated as a function of metal (Cr, Mo, or W) and ligand (-OH, -OCH3). A number of fundamental thermodynamic properties of the anions and corresponding neutrals were predicted on the basis of the theoretical calculations. The calculations indicate high O-H bond dissociation energies for MO2(OR)(O-H) (R = H, CH3) and MO3(O-H), consistent with their high Bronsted acidities (just below that of H2SO4 in the gas phase) and the high ionization energies of their conjugate base anions. This suggests that the corresponding radicals should readily abstract H atoms from organic molecules. PMID- 16366628 TI - Dissociative photodetachment dynamics of solvated iodine cluster anions. AB - Photoelectron-photofragment coincidence spectroscopy of I- (CO2), I- (NH3), I- (H2O), I- (C6H5NH2), and I- (C6H5OH) clusters was used to study the dissociative photodetachment (DPD) dynamics at 257 nm. Photodetachment from all five clusters was observed to yield bound neutral clusters as well as the DPD products of the iodine atom and the molecular solvent. Photoelectron images and kinetic energy spectra were recorded in coincidence with both the translational energy released between dissociating neutral products and stable neutral clusters. The variation of the photoelectron angular distributions in the clusters was measured, revealing significant perturbations relative to I- for I- (H2O) and I- (C6H5NH2). Product branching ratios for stable versus dissociative photodetachment and photodetachment to the I(2P(3/2)) and I(2P(1/2)) states are reported. The measurements reveal a dependence of the DPD dynamics on the final spin-orbit state of iodine in the cases of I- (C6H5NH2) and I- (CO2) and a threshold detachment process in I- (C6H5NH2). PMID- 16366629 TI - Size-dependent carbon monoxide adsorption on neutral gold clusters. AB - We report on experiments probing the reactivity of neutral Au(n) clusters, n = 9 68, with carbon monoxide. The gold clusters are produced in a pulsed laser vaporization cluster source, operated at room temperature (RT) or at liquid nitrogen temperature (LNT), pass through a low-pressure reaction cell containing CO gas, and are subsequently laser ionized. The reaction probabilities are determined by recording mass abundance spectra with time-of-flight mass spectrometry. The main observations are a strong temperature dependence and a remarkable size dependence. Upon cooling of the cluster source to LNT, the reactivity increases substantially. At LNT, the reaction probabilities for Au(n) with the first CO molecule are about a factor 10 higher than at RT. Moreover, adsorption of two, three, and even four CO molecules is observed, in contrast to RT clusters which at most adsorb one CO molecule. This temperature dependence is related to the lifetime of the cluster-molecule complexes, being much longer for cold clusters. The observed striking size dependence is similar at both temperatures and is discussed in terms of the electronic structure effects. PMID- 16366630 TI - Empty level structure in phenyl and benzyl isocyanates. AB - The energies of the lowest-lying anion states of phenyl (C6H5N=C=O) and benzyl (C6H5CH2N=C=O) isocyanates have been determined experimentally in the gas phase for the first time using electron transmission spectroscopy (ETS), and their localization properties have been evaluated using HF/6-31G, MP2/6-31G*, and B3LYP/6-31G* calculations. The lowest-lying anion state of phenyl isocyanate, mainly of benzene ring character but with some contribution also from the N=C=O pi-system, lies at significantly higher energy than that of other benzenes substituted by pi-functionals, such as benzaldehyde or styrene. The scaling with the use of suitable empirical equations of the virtual orbital energies (VOEs) for orbitals with predominantly pi*(ring) character calculated for the neutral state molecules leads to vertical attachment energies (VAEs) which closely correspond to those determined experimentally, whereas those calculated for the predominantly pi*(CO) and pi*(NC) orbitals (3rd and 4th LUMO, respectively) are significantly different from the corresponding measured values notwithstanding the fact that the calculations reproduce the shortening of the N=C and C=O double bonds. PMID- 16366631 TI - Very fast tunneling in the early stage of reaction dynamics. AB - The difference between quantum and classical survival probabilities for molecular dissociation dynamics in the time domain, which arises mainly from quantum mechanical tunneling, has interesting characteristics that are not noticed through the counterpart in energy domain. It is shown that the early stage undergoes a fast tunneling, while the later stage is characterized with a long lasting slow tunneling. The mechanism of this behavior is analyzed in terms of a quasi-semiclassical theory featuring the geometrical distribution of the so called tunneling points. In particular, the role of dynamical tunneling is discussed as a phenomenon that typifies the time dependence of tunneling dynamics. It is predicted that these tunneling characteristics will be reflected in the isotope effect and should be experimentally observable. PMID- 16366632 TI - FTIR studies of O(3P) atom reactions with CSe2, SCSe, and OCSe. AB - The overall rate coefficients of the reactions of CSe2, SCSe, and OCSe with O(3P) atom have been determined to be k(CSe2) = (1.4 +/- 0.2) x 10(-10) cm3 molecule( 1) s(-1), k(SCSe) = (2.8 +/- 0.3) x 10(-11) cm3 molecule(-1) s(-1), and k(OCSe) = (2.4 +/- 0.3) x 10(-11) cm3 molecule(-1) s(-1) at 301-303 K using Fourier transform infrared (FTIR) absorption spectroscopy. The measurements have been accomplished by calibrating against the literature value of the rate coefficient for O(3P) with CS2 (4 x 10(-12) cm3 molecule(-1) s(-1)). A product channel giving OCSe in 32.0 +/- 4.2% yield has been found for the O + CSe2 reaction. Although CO was also detected, its generation could be attributed to subsequent reactions of OCSe with O atoms. The corresponding reaction for O + SCSe gives OCS and OCSe as observable products, with their yields given as 32.2 +/- 4.5 and 30.2 +/- 3.3%, respectively. Computational studies using UB3LYP/aug-cc-PVTZ methods have been used particularly to determine the reaction pathways for the channels in which OCS or OCSe is produced. PMID- 16366633 TI - Rate constant and activation energy measurement for the reaction of atomic hydrogen with thiocyanate and azide in aqueous solution. AB - Arrhenius parameters for the reaction of hydrogen atoms with azide and thiocyanate in aqueous solution have been determined using electron pulse radiolysis and electron paramagnetic resonance free induction decay attenuation measurements. Absolute values for SCN-, N3(-), and HN3 were well-described over the temperature range of 9-81 degrees C by the equations log k5 = (12.03 +/- 0.12) - [(21.05 +/- 0.66 kJ mol(-1))/2.303RT], log k10 = (12.75 +/- 0.21) - [(18.43 +/- 1.22 kJ mol(-1))/2.303RT], and log k15 = (11.59 +/- 0.12) - [(21.44 +/- 0.69 kJ mol(-1))/2.303RT], corresponding to room temperature (22 degrees C) rate constants of (2.07 +/- 0.03) x 10(8), (3.15 +/- 0.08) x 10(9), and (6.31 +/- 0.05) x 10(7) M(-1) s(-1) and activation energies for these chemicals of 21.05 +/ 0.66, 18.4 +/- 1.2, and 21.44 +/- 0.69 kJ mol(-1), respectively. The similarity of these three measured activation energies, taken together with the available information on reaction products, suggests a similar reaction mechanism, which is proposed to be an initial hydrogen atom adduct formation in these molecules, followed by single bond breakage. PMID- 16366634 TI - Atmospheric chemistry of dimethyl phosphonate, dimethyl methylphosphonate, and dimethyl ethylphosphonate. AB - Rate constants for the reactions of OH radicals and NO3 radicals with dimethyl phosphonate [DMHP, (CH3O)2P(O)H], dimethyl methylphosphonate [DMMP, (CH3O)2P(O)CH3], and dimethyl ethylphosphonate [DMEP, (CH3O)2P(O)C2H5] have been measured at 296 +/- 2 K and atmospheric pressure using relative rate methods. The rate constants obtained for the OH radical reactions (in units of 10(-12) cm3 molecule(-1) s(-1)) were as follows: DMHP, 4.83 +/- 0.25; DMMP, 10.4 +/- 0.6; and DMEP, 17.0 +/- 1.0, with a deuterium isotope effect of k(OH + DMMP)/k(OH + DMMP d9) = 4.8 +/- 1.2. The rate constants obtained for the NO3 radical reactions (in units of 10(-16) cm3 molecule(-1) s(-1)) were as follows: DMHP, < 1.4; DMMP, 2.0 +/- 1.0; and DMEP, 3.4 +/- 1.4. Upper limits to the rate constants for the O3 reactions of < 8 x 10(-20) cm3 molecule(-1) s(-1) for DMHP and < 6 x 10(-20) cm3 molecule(-1) s(-1) for DMMP and DMEP were determined. Products of the reactions of OH radicals with DMHP, DMMP, and DMEP were investigated in situ using atmospheric pressure ionization mass spectrometry (API-MS) and, for the DMMP and DMEP reactions, Fourier transform infrared (FT-IR) spectroscopy. API-MS analyses showed the formation of products of molecular weight 96 and 126, attributed to CH3OP(O)(H)OH and (CH3O)2P(O)OH, respectively, from DMHP; of molecular weight 110, attributed to CH3OP(O)(CH3)OH, from DMMP; and of molecular weight 124 and 126, attributed to CH3OP(O)(C2H5)OH and (CH3O)2P(O)OH, respectively, from DMEP. FT-IR analyses showed formation (values given are % molar yields) of the following: from DMMP, CO, 54 +/- 6; CO2, 5 +/- 1 in dry air; HCHO, 3.9 +/- 0.7; HC(O)OH, < 1.4 in dry air; RONO2, approximately 4; and formate ester, approximately 8; and from DMEP, CO, 50 +/- 7; CO2, 11 +/- 4; CH3CHO, 18 +/- 8; HCHO, < 7; HC(O)OH, < 6; RONO2, < or = 5; and formate ester, 5.0 +/- 1.5. Possible reaction mechanisms are discussed. PMID- 16366635 TI - Atmospheric chemistry of propionaldehyde: kinetics and mechanisms of reactions with OH radicals and Cl atoms, UV spectrum, and self-reaction kinetics of CH3CH2C(O)O2 radicals at 298 K. AB - The kinetics and mechanism of the reactions of Cl atoms and OH radicals with CH3CH2CHO were investigated at room temperature using two complementary techniques: flash photolysis/UV absorption and continuous photolysis/FTIR smog chamber. Reaction with Cl atoms proceeds predominantly by abstraction of the aldehydic hydrogen atom to form acyl radicals. FTIR measurements indicated that the acyl forming channel accounts for (88 +/- 5)%, while UV measurements indicated that the acyl forming channel accounts for (88 +/- 3)%. Relative rate methods were used to measure: k(Cl + CH3CH2CHO) = (1.20 +/- 0.23) x 10(-10); k(OH + CH3CH2CHO) = (1.82 +/- 0.23) x 10(-11); and k(Cl + CH3CH2C(O)Cl) = (1.64 +/- 0.22) x 10(-12) cm3 molecule(-1) s(-1). The UV spectrum of CH3CH2C(O)O2, rate constant for self-reaction, and rate constant for cross-reaction with CH3CH2O2 were determined: sigma(207 nm) = (6.71 +/- 0.19) x 10(-18) cm2 molecule(-1), k(CH3CH2C(O)O2 + CH3CH2C(O)O2) = (1.68 +/- 0.08) x 10(-11), and k(CH3CH2C(O)O2 + CH3CH2O2) = (1.20 +/- 0.06) x 10(-11) cm3 molecule(-1) s(-1), where quoted uncertainties only represent 2sigma statistical errors. The infrared spectrum of C2H5C(O)O2NO2 was recorded, and products of the Cl-initiated oxidation of CH3CH2CHO in the presence of O2 with, and without, NO(x) were identified. Results are discussed with respect to the atmospheric chemistry of propionaldehyde. PMID- 16366636 TI - Protonation thermochemistry of selected hydroxy- and methoxycarbonyl molecules. AB - The gas-phase basicities of a representative set of hydroxy- and methoxycarbonyl compounds (hydroxyacetone, 1, 3-hydroxybutanone, 2, 3-hydroxy-3-methylbutanone, 3, 1-hydroxy-2-butanone, 4, 4-hydroxy-2-butanone, 5, 5-hydroxy-2-pentanone, 6, methoxyacetone, 7, 3-methoxy-2-butanone, 8, 4-methoxy-2-butanone, 9, and 5 methoxy-2-pentanone, 10) were experimentally determined by the equilibrium method using Fourier transform ion cyclotron resonance and high-pressure mass spectrometry techniques. The latter method allows the measurement of proton transfer equilibrium constants at various temperatures and thus the estimate of both the proton affinities and the protonation entropies of the relevant species. Quantum chemical calculations at the G3 and the B3LYP/6-311+G(3df,2p)//6-31G(d) levels of theory were undertaken in order to find the most stable structures of the neutrals 1-10 and their protonated forms. Conformational and vibrational analyses have been done with the aim of obtaining a theoretical estimate of the protonation entropies. PMID- 16366637 TI - Excited state properties of 7-hydroxy-4-methylcoumarin in the gas phase and in solution. A theoretical study. AB - TDDFT/B3LYP and RI-CC2 calculations with different basis sets have been performed for vertical and adiabatic excitations and emission properties of the lowest singlet states for the neutral (enol and keto), protonated and deprotonated forms of 7-hydroxy-4-methylcoumarin (7H4MC) in the gas phase and in solution. The effect of 7H4MC-solvent (water) interactions on the lowest excited and fluorescence states were computed using the Polarizable Continuum Method (PCM), 7H4MC-water clusters and a combination of both approaches. The calculations revealed that in aqueous solution the pi pi* energy is the lowest one for excitation and fluorescence transitions of all forms of 7H4MC studied. The calculated excitation and fluorescence energies in aqueous solution are in good agreement with experiment. It was found that, depending on the polarity of the medium, the solvent shifts vary, leading to a change in the character of the lowest excitation and fluorescence transition. The dipole-moment and electron density changes of the excited states relative to the ground state correlate with the solvation effect on the singlet excited states and on transition energies, respectively. The calculations show that, in contrast to the ground state, the keto form has a lower energy in the pi pi* state as compared to enol, demonstrating from this point of view the energetic possibility of proton transfer from the enol to the keto form in the excited state. PMID- 16366638 TI - Density functional theoretical investigation on influence of heterosubstitution and benzannelation on the thermal 6pi electrocyclization of cis cyclononatetraene. AB - Thermal 6pi electrocyclization of cyclononatetraene (CNT), its hetero-substituted analogues, and its benzannelated derivatives have been investigated by using the B3LYP method employing 6-31G* and 6-311+G** basis sets. The results indicate that heterosubstitution and benzannelation influence the rate of cyclization. Nucleus independent chemical shifts (NICS), conceptual density functional theory (DFT) based reactivity descriptors, group electronegativity values, and barriers to planarity provide complementary evidence for the predicted rate of cyclization. The available experimental data are in good agreement with the computed values. PMID- 16366639 TI - Solvation properties of N-substituted cis and trans amides are not identical: significant enthalpy and entropy changes are revealed by the use of variable temperature 1H NMR in aqueous and chloroform solutions and ab initio calculations. AB - The cis/trans conformational equilibrium of N-methyl formamide (NMF) and the sterically hindered tert-butylformamide (TBF) was investigated by the use of variable temperature gradient 1H NMR in aqueous solution and in the low dielectric constant and solvation ability solvent CDCl3 and various levels of first principles calculations. The trans isomer of NMF in aqueous solution is enthalpically favored relative to the cis (deltaH(o) = -5.79 +/- 0.18 kJ mol(-1)) with entropy differences at 298 K (298 x deltaS(o) = -0.23 +/- 0.17 kJ mol(-1)) playing a minor role. The experimental value of the enthalpy difference strongly decreases (deltaH(o) = -1.72 +/- 0.06 kJ mol(-1)), and the contribution of entropy at 298 K (298 x deltaS(o) = -1.87 +/- 0.06 kJ mol(-1)) increases in the case of the sterically hindered tert-butylformamide. The trans isomer of NMF in CDCl3 solution is enthalpically favored relative to the cis (deltaH(o) = -3.71 +/ 0.17 kJ mol(-1)) with entropy differences at 298 K (298 x deltaS(o) = 1.02 +/- 0.19 kJ mol(-1)) playing a minor role. In the sterically hindered tert butylformamide, the trans isomer is enthalpically disfavored (deltaH(o) = 1.60 +/ 0.09 kJ mol(-1)) but is entropically favored (298 x deltaS(o) = 1.71 +/- 0.10 kJ mol(-1)). The results are compared with literature data of model peptides. It is concluded that, in amide bonds at 298 K and in the absence of strongly stabilizing sequence-specific inter-residue interactions involving side chains, the free energy difference of the cis/trans isomers and both the enthalpy and entropy contributions are strongly dependent on the N-alkyl substitution and the solvent. The significant decreasing enthalpic benefit of the trans isomer in CDCl3 compared to that in H2O, in the case of NMF and TBF, is partially offset by an adverse entropy contribution. This is in agreement with the general phenomenon of enthalpy versus entropy compensation. B3LY/6-311++G** and MP2/6-311++G** quantum chemical calculations confirm the stability orders of isomers and the deltaG decrease in going from water to CHCl3 as solvent. However, the absolute calculated values, especially for TBF, deviate significantly from the experimental values. Consideration of the solvent effects via the PCM approach on NMF x H2O and TBF x H2O supermolecules improves the agreement with the experimental results for TBF isomers, but not for NMF. PMID- 16366640 TI - Influence of multi-atom bridging ligands on the electronic structure and magnetic properties of homodinuclear titanium molecules. AB - The electronic structure and magnetic properties of homodinuclear titanium(III) molecules with bridging ligands from groups 14, 15, and 16 are examined. Single- and multireference methods with triple-zeta plus polarization basis sets are employed. Dynamic electron correlation effects are included via second-order multireference perturbation theory. Isotropic interaction parameters are calculated, and two of the complexes studied are predicted to be ferromagnetic based on multireference second-order perturbation (MRMP2) theory, using the TZVP(fg) basis set. Zero-field splitting parameters are determined using spin orbit coupling obtained from complete active space (CAS) self-consistent field (SCF) and multiconfigurational quasi-degenerate perturbation theory (MCQDPT) wave functions. Three Breit-Pauli-based spin coupling methods were employed: full Breit-Pauli (HSO2), the partial two-electron method (P2E), and the semiempirical one-electron method (HSO1). PMID- 16366641 TI - The mechanism of photochemical 1,3-silyl migration of allylsilane. AB - The photochemical reaction mechanisms of model compounds for 4-tert-butyl-1-(4 phenylphenyl)-1-(1,1-dimethylallyl)silacyclohexane are investigated using a complete active space comprised of six electrons in six orbitals with the standard 6-31G(d) basis set. It is concluded that the stereochemistry in the photochemical 1,3-silyl migrations of allylsilanes has a retention preference, in accord with the Woodward-Hoffmann rules. The calculated conical intersection (CI) structure suggests a dissociation path to radicals in addition to a 1,3-shift path. The bulkiness and rigidness of a silacyclohexane moiety does not affect the stereochemistry, but a slightly elongated Si-C bond length in the CI structure would promote the dissociation path. PMID- 16366642 TI - B3LYP and MP2 calculations of the enthalpies of hydrogen-bonded complexes of methanol with neutral bases and anions: comparison with experimental data. AB - To perfect a method for building a theoretical hydrogen-bond basicity scale, the enthalpy of hydrogen bonding between methanol and thirteen neutral and anionic bases (MeOH, MeNH2, Me2NH, Et2NH, Me3N, Et3N, Br-, CN-, SH-, Cl-, HCOO-, MeO-, F ) was calculated by DFT and ab initio methods. The theoretical results were compared to selected experimental ones. It appears that B3LYP/6-31+G(d,p) calculations are satisfactory for optimizing the geometry of complexes and giving a general order of basicity. However, they are deficient for reproducing the large effect of alkyl groups on the hydrogen-bond basicity of amines. This deficiency is explained by intermolecular perturbation theory calculations, which show that the alkylation of nitrogen dramatically increases the dispersion energy component not taken into account by the B3LYP functional. Of the methods considered, only MP2/aug-cc-pVTZ calculations are capable of reproducing the binding enthalpy within the experimental error for the first-row acceptor atoms N, O, and F, and of accounting for dispersion effects created by alkylation at the hydrogen-bond acceptor site. PMID- 16366643 TI - Singlet and triplet products of the geminate recombination of a radical pair with a single magnetic nucleus (I = 1/2). AB - The double-channel recombination and separation of the photochemically created singlet radical pair is investigated, taking into account the spin conversion in a zero magnetic field and the arbitrary initial distance between the radicals. The quantum yields of the singlet and triplet products and the free radicals production are found analytically, assuming that the recombination of the diffusing radicals occurs at contact. All the yields are related to the singlet and triplet populations of the recombining radical pair, subjected to spin conversion and contact exchange interaction. The general analytical expressions for the quantum yields are specified for the particular limits of the weak and strong exchange. They are greatly simplified in the case of polar solvents, especially at the contact start. A close similarity is obtained with the results of a previously developed incoherent model of spin conversion, provided that the conversion rate is appropriately related to the hyperfine coupling constant. PMID- 16366644 TI - Accurate ab initio binding energies of alkaline earth metal clusters. AB - The effects of basis set superposition error (BSSE) and core-correlation on the electronic binding energies of alkaline earth metal clusters Y(n) (Y = Be, Mg, Ca; n = 2-4) at the Moller-Plesset second-order perturbation theory (MP2) and the single and double coupled cluster method with perturbative triples correction (CCSD(T)) levels are examined using the correlation consistent basis sets cc-pVXZ and cc-pCVXZ (X = D, T, Q, 5). It is found that, while BSSE has a negligible effect for valence-electron-only-correlated calculations for most basis sets, its magnitude becomes more pronounced for all-electron-correlated calculations, including core electrons. By utilizing the negligible effect of BSSE on the binding energies for valence-electron-only-correlated calculations, in combination with the negligible core-correlation effect at the CCSD(T) level, accurate binding energies of these clusters up to pentamers (octamers in the case of the Be clusters) are estimated via the basis set extrapolation of ab initio CCSD(T) correlation energies of the monomer and cluster with only the cc-pVDZ and cc-pVTZ sets, using the basis set and correlation-dependent extrapolation formula recently devised. A comparison between the CCSD(T) and density functional theory (DFT) binding energies is made to identify the most appropriate DFT method for the study of these clusters. PMID- 16366645 TI - Theoretical investigation of the dihydrogen bond linking MH2 with HCCRgF (M = Zn, Cd; Rg = Ar, Kr). AB - An ab initio computational study of the properties of four linear dihydrogen bonded complexes pairing MH2 (M = Zn, Cd) with HCCRgF (Rg = Ar, Kr) was undertaken at the MP2/DGDZVP level of theory. The calculated complexation energies of the linear complexes vary between 6.5 kJ/mol for M = Zn to 8.5 kJ/mol for M = Cd. Equilibrium interatomic H...H distances are roughly 2.07 A for all four complexes. The red shifts of the H-C stretching frequency of HCCRgF correlate nicely with the interaction energies. PMID- 16366646 TI - Natural energy decomposition analysis: extension to density functional methods and analysis of cooperative effects in water clusters. AB - Natural energy decomposition analysis (NEDA) is a method for partitioning molecular interaction energies into physically meaningful components, including electrical interaction, charge transfer, and core repulsions. The method is a numerically stable procedure that was originally developed for analyzing Hartree Fock (HF) wave functions based on the localized orbital description of natural bond orbital analysis. In this work, we extend NEDA to treat charge densities from density functional theory (DFT) calculations, replacing the intermolecular exchange (EX) component of the HF analysis with an exchange-correlation (XC) component. DFT/NEDA is applied to hydrogen bonding interactions and cooperative effects in water clusters. Electrical interactions and charge transfer contribute importantly to hydrogen bonding. Comparison of HF and DFT results reveals that the exchange and correlation effects of DFT slightly enhance the extent of charge transfer and core repulsions in the water clusters. Cooperative stabilization of the cyclic water trimer and tetramer is considered by performing a many-body expansion of the interaction energy. Natural energy decomposition analysis of this expansion suggests that charge transfer is the leading source of cooperative stabilization. Polarization effects have only marginal influence on cooperativity. PMID- 16366647 TI - Radical-molecule reactions HCO/HOC + C2H2: mechanistic study. AB - A detailed computational study is performed on the unknown radical-molecule reactions between HCO/HOC and acetylene (C2H2) at the CCSD(T)/6 311G(2d,p)//B3LYP/6-311G(d,p)+ZPVE, Gaussian-3//B3LYP/6-31G(d), and Gaussian 3//MP2(full)/6-31G(d) levels. For the HCO + C2H2 reaction, the most favorable pathway is direct C-addition forming the intermediate HC=CHCH=O followed by a 1,3 H-shift leading to H2C=CHC=O, which finally dissociates to the product C2H3 + CO. The overall reaction barrier is 13.8, 10.5, and 11.3 kcal/mol, respectively, at the three levels. The quasi-direct H-donation process to produce C2H3 + CO with barriers of 14.0, 14.1, and 14.1 kcal/mol is less competitive. Thus only at higher temperatures could the HCO + C2H2 reaction play a role. In contrast, the HOC + C2H2 reaction can barrierlessly generate C2H3 + CO via the quasi-direct H donation mechanism proceeding via a prereactive complex with OH...C2 hydrogen bonding. This is suggestive of the potential importance of the HOC + C2H2 reaction in both combustion and interstellar processes. However, the direct C addition channel is much less competitive. For both reactions, the possible formation of the intriguing interstellar molecules propadiene and propynal is also discussed. The present theoretical study represents the first attempt to probe the reaction mechanism between HOC and pi-systems. Future laboratory investigations on both reactions (particularly HOC + C2H2) are recommended. PMID- 16366648 TI - Triangular halogen trimers. A DFT study of the structure, cooperativity, and vibrational properties. AB - Triangular halogen trimers (RX)3, where X = Br, I and R represents H, H3C, H2FC, HF2C, F3C, CH2=CH, CH[triple bond]C, and Ph, have been investigated using the density functional theory in the Perdew, Burke, and Ernzerhof method. We report herein the optimized geometries of the stable structures, their vibrational frequencies, and binding energies with the two- and three-body terms. All trimer structures possess a cyclic array of halogen atoms in the type II approach by virtue of the nonspherical atomic charge distribution around the halogens. The Br...Br interactions in trimers are very weak, whereas the I...I interactions in trimers are relatively stronger. While all bromine trimers and most of iodine trimers are predicted to be noncooperative, three of iodine trimers show weak cooperativity. The analysis of vibration modes reveals that all halogen trimers exhibit no especially remarkable frequency shifts. It is also shown that the electrostatic contribution plays a major role in the halogen...halogen interactions in halogen trimers. In contrast to bromine trimers, the relative contribution of charge-transfer component to the halogen...halogen interactions becomes more important for iodine trimers. PMID- 16366649 TI - A theoretical study of 31P NMR chemical shielding models for concentrated phosphoric acid solution. AB - Calculations on the hydrates, dimer, and trimer of phosphoric acid were carried out in an effort to obtain a viable model of the phosphorus NMR chemical shielding in 85% phosphoric acid solution. The theoretical approaches used the gauge-including-atomic-orbital (GIAO) 6-311+G(nd,p) basis set at both scaled density functional theory (sB3LYP) and estimated infinite order Moller-Plesset (EMPI) approaches and with the aug-cc-pvtz basis in the sB3LYP approach. Shieldings and hydrogen bonding stabilization energies are similar in the three approaches and indicate that the faster sB3LYP/6-311+G(nd,p) approach can be used with larger systems. The changes in shielding compared to the isolated species are small and suggest that the undissociated acid dihydrate could serve as a model entity for modeling the phosphorus shielding in concentrated phosphoric acid solution. PMID- 16366650 TI - Quantum chemical study of the mechanism of reaction between NH (X 3sigma-) and H2, H2O, and CO2 under combustion conditions. AB - Reactions of ground-state NH (3sigma-) radicals with H2, H2O, and CO2 have been investigated quantum chemically, whereby the stationary points of the appropriate reaction potential energy surfaces, that is, reactants, products, intermediates, and transition states, have been identified at the G3//B3LYP level of theory. Reaction between NH and H2 takes place via a simple abstraction transition state, and the rate coefficient for this reaction as derived from the quantum chemical calculations, k(NH + H2) = (1.1 x 10(14)) exp(-20.9 kcal mol(-1)/RT) cm3 mol(-1) s(-1) between 1000 and 2000 K, is found to be in good agreement with experiment. For reaction between triplet NH and H2O, no stable intermediates were located on the triplet reaction surface although several stable species were found on the singlet surface. No intersystem crossing seam between triplet NH + H2O and singlet HNO + H2 (the products of lowest energy) was found; hence there is no evidence to support the existence of a low-energy pathway to these products. A rate coefficient of k(NH + H2O) = (6.1 x 10(13)) exp(-32.8 kcal mol(-1)/RT) cm3 mol(-1) s(-1) between 1000 and 2000 K for the reaction NH (3sigma-) + H2O --> NH2 (2B) + OH (2pi) was derived from the quantum chemical results. The reverse rate coefficient, calculated via the equilibrium constant, is in agreement with values used in modeling the thermal de-NO(x) process. For the reaction between triplet NH and CO2, several stable intermediates on both triplet and singlet reaction surfaces were located. Although a pathway from triplet NH + CO2 to singlet HNO + CO involving intersystem crossing in an HN-CO2 adduct was discovered, no pathway of sufficiently low activation energy was discovered to compare with that found in an earlier experiment [Rohrig, M.; Wagner, H. G. Proc. Combust. Inst. 1994, 25, 993.]. PMID- 16366651 TI - Theoretical study of the stability of lithium atoms in alpha-rhombohedral boron. AB - The stability of lithium atoms in alpha-rhombohedral boron was investigated by first-principles calculations of total energies and molecular dynamics (MD) simulations. In the case of a low concentration (1.03 at. %), Li at the center of the icosahedral B12 site (the I-site) had a negative binding energy, which suggests Li at the I-site is unstable. However, MD simulations at temperatures below 750 K indicated that Li is still confined in the B12 cage under these conditions, which means Li at the I-site is metastable. Over 800 K, Li began to move away from the B12 site and settled at the tetrahedral site (the T-site) or at the octahedral site (the O-site). Li at the T-site also had a negative binding energy, but MD simulations indicated it was metastable up to 1400 K and did not move to other sites. Li at the O-site was energetically the most favorable, having a positive binding energy. In the case of a high concentration (7.69 at. %), the I-site changed to an unstable saddle point. At this concentration, the T site was metastable and the O-site became the most stable. In MD simulations at 1400 K, Li atoms at the O-site never jumped to other sites regardless of concentration. Considering these facts, the diffusion coefficient of Li in alpha rhombohedral boron would have to be very small below 1400 K. PMID- 16366652 TI - CLOPPA-IPPP analysis of electronic mechanisms of intermolecular (1h)J(A,H) and (2h)J(A,D) spin-spin coupling constants in systems with D-H...A hydrogen bonds. AB - The electronic origin of intermolecular (2h)J(A,D) and (1h)J(A,H) couplings is discussed by means of the CLOPPA-IPPP approach in several model complexes with D H...A hydrogen bonds. It is found that the origin of these couplings is mainly due to the interaction between the acceptor sigma lone pair and vacant molecular orbitals localized in the D-H...A moiety, regardless of the donor and acceptor nuclei. The problem of the larger absolute value of (2h)J(A,D) compared to (1h)J(A,H) is also addressed. PMID- 16366653 TI - Ab initio characterization of electron transfer coupling in photoinduced systems: generalized Mulliken-Hush with configuration-interaction singles. AB - To calculate electronic couplings for photoinduced electron transfer (ET) reactions, we propose and test the use of ab initio quantum chemistry calculation for excited states with the generalized Mulliken-Hush (GMH) method. Configuration interaction singles (CIS) is proposed to model the locally excited (LE) and charge-transfer (CT) states. When the CT state couples with other high lying LE states, affecting coupling values, the image charge approximation (ICA), as a simple solvent model, can lower the energy of the CT state and decouple the undesired high-lying local excitations. We found that coupling strength is weakly dependent on many details of the solvent model, indicating the validity of the Condon approximation. Therefore, a trustworthy value can be obtained via this CIS GMH scheme, with ICA used as a tool to improve and monitor the quality of the results. Systems we tested included a series of rigid, sigma-linked donor-bridge acceptor compounds where "through-bond" coupling has been previously investigated, and a pair of molecules where "through-space" coupling was experimentally demonstrated. The calculated results agree well with experimentally inferred values in the coupling magnitudes (for both systems studied) and in the exponential distance dependence (for the through-bond series). Our results indicate that this new scheme can properly account for ET coupling arising from both through-bond and through-space mechanisms. PMID- 16366654 TI - Effects of peripheral substituents and axial ligands on the electronic structure and properties of cobalt porphyrins. AB - The effects of peripheral substituents and axial ligands (L) on the electronic structure and properties of cobalt tetraphenylporphyrin (CoTPP) have been studied using DFT methods. Various density functionals were tested, and the ground state of each system was determined by considering several possible low-lying states. The ground states of the fully fluorinated CoTPPF28(L)2 complexes with L = THF, Py, and Im were identified to be high-spin (4E(g)) by the meta-GGA functional tau HCTH, which contains the kinetic energy density tau, in agreement with experimental measurements. All the pure GGA functionals, including the recently developed mPBE, OPBE, and HCTH/407, show more or less overestimation of the relative energies of the high-spin states. The energy gap between the 2A(1g) and 4E(g) states is insignificant (approximately 0.1 eV) and varies in the order L = Py < L = THF < L = Im. The results and their trend are consistent with 19F NMR studies which show partial population of the 4E(g) state in CoTPPF28(THF)2 and CoTPPF28(Py)2 and a complete conversion to the high-spin state in CoTPPF28(1 MeIm)2. Upon coordination by two very strong field axial CO ligands, CoTPPF28(CO)2 becomes low-spin, as in unligated CoTPPF(x). The influence of the peripheral substituents and axial ligands on the ionization potentials, electron affinities, and CoTPPF(x)-(L)2 binding strength was also investigated in detail. PMID- 16366655 TI - Theoretical study of (CH...C)- hydrogen bonds in CH(4-n)X(n) (X = F, Cl; n = 0, 1, 2) systems complexed with their homoconjugate and heteroconjugate carbanions. AB - This work deals with a theoretical study of the (CH...C)- hydrogen bonds in CH4, CH3X, and CH2X2 (X = F, Cl) complexed with their homoconjugate and heteroconjugate carbanions. The properties of the complexes are calculated with the B3LYP method using the 6-311++G(d,p) or 6-311++G(2df,2p) basis sets. The deprotonation enthalpies (DPE) of the CH bond or the proton affinities of the carbanions (PA(C-) are calculated as well. All the systems with the exception of the CH4...CHCl2(-) one are characterized by a double minimum potential. In some of the complexes, the (CH(b)...C)- hydrogen bond is linear. In other systems, such as CH3F...CH2F- and CH3F...CHF2(-), there is a large departure from linearity, the systems being stabilized by electrostatic interactions between the nonbonded H of the neutral molecule and the F atom of the carbanion. In the transition state, the (CH(b)...C)- bond is linear, and there is a large contraction of the intermolecular C...C distance. The binding energies vary within a large range, from -1.4 to -11.1 kcal mol(-1) for the stable complexes and -8.6 to -44.1 kcal mol(-1) for the metastable complexes. The energy barriers to proton transfer are between 5 and 20 kcal mol(-1) for the heteroconjugate systems and between 3.8 and 8.3 kcal mol(-1) for the homoconjugate systems. The binding energies of the linear complexes depend exponentially on 1.5DPE - PA(C-), showing that the proton donor is more important than the proton acceptor in determining hydrogen bond strength. The NBO analysis indicates an important electronic reorganization in the two partners. The elongations of the CH bond resulting from the interaction with the carbanion depend on the occupation of the sigma*(CH(b)) antibonding orbitals and on the hybridization of the C bonded to H(b). The frequency shifts of the nu(CH)(A1) stretching vibration range between 15 and 1150 cm(-1). They are linearly correlated to the elongation of the CH(b) bond. PMID- 16366656 TI - Study of the dative bond in 2-aminoethoxydiphenyl borate at various levels of theory: another poor performance of the B3LYP method for B-N dative bonds. AB - Aminoethoxydiphenyl borate (2-APB), 1, is a potent inhibitor of store-operated calcium entry channels (SOCCs). Other SOCC inhibitors are being investigated as promising pharmacological agents for a variety of conditions. Though toxic, 2-APB could be useful in the development of additional inhibitors, but its preferred binding structure must first be determined. Thus, we performed ab initio calculations to study the conformers and the strength of the dative bond of 2 APB. As a first step, we performed a series of computations at various levels of theory. We obtained vastly different dissociation energies for the dative bond depending on whether we used MP2 or B3LYP (7-10 kcal/mol different). This discrepancy has previously been observed for other B-N dative bonds by Gilbert, who found that the MP2 values were in much better agreement with experimental values (Gilbert, T. M. J. Phys. Chem. A 2004, 108, 2550-2554). Since we lacked experimental data for comparison, we performed CCSD(T) calculations and found them to have similar results to those from MP2. Thus, we conclude that MP2 is more accurate for 2-APB. The dissociation free energy at the MP2 level is 7 kcal/mol and indicates that the dative bond conformer will be the predominant structure in the gas phase. The dissociation energy is comparatively low due to the electron donation from the oxygen atom to the boron atom and due to the ring strain in the dative bond conformer. PMID- 16366657 TI - Hydrolysis of fluorosilanes: a theoretical study. AB - Hydrolysis and condensation of simple trifluorosilanes, HSiF3 and MeSiF3, was studied by quantum mechanical methods. Hydrolysis of fluorosilanes is highly endothermic. The Gibbs free energy of the first reaction step in the gas phase is 31.4 kJ/mol, which corresponds to an equilibrium constant of 10(-6). Hydrolysis of the subsequent fluorine atoms in trifluorosilanes is thermodynamically more unfavorable than the first step of substitution. No significant difference in thermodynamics of hydrolysis was found between HSiF3 and MeSiF3. The activation energy for hydrolysis by a water dimer is significantly lower than that for hydrolysis by a water monomer. The former reaction is also less unfavorable thermodynamically, due to a high binding energy of the HF-H2O complex formed as a product of hydrolysis. Self-consistent reaction field (SCRF) calculations show that hydrolysis of trifluorosilanes in aqueous medium has lower activation energy than in the gas phase. It is also thermodynamically less unfavorable, due to better solvation of the products. Homofunctional condensation of HSiF2OH is thermodynamically favored. The equilibrium mixture for hydrolysis/condensation of RSiF3 in water is predicted to contain ca. 2.3% disiloxane (HF2Si)2O, if 100-fold excess of water relative to silane is assumed. Further hydrolysis of (HF2Si)2O is negligible. The thermodynamics of fluorosilane hydrolysis contrasts with that of chlorosilanes, where both hydrolysis and condensation are strongly favorable. Moreover, in the case of trichlorosilanes each subsequent hydrolysis step is more facile, leading to the product of full hydrolysis, RSi(OH)3. PMID- 16366658 TI - Direct dynamics study on the hydrogen abstraction reaction CH2O + HO2 --> CHO + H2O2. AB - We present a direct ab initio dynamics study on the hydrogen abstraction reaction CH2O + HO2 --> CHO + H2O2, which is predicted to have four possible reaction channels caused by different attacking orientations of HO2 radical to CH2O. The structures and frequencies at the stationary points and the points along the minimum energy paths (MEPs) of the four reaction channels are calculated at the B3LYP/cc-pVTZ level of theory. Energetic information of stationary points and the points along the MEPs is further refined by means of some single-point multilevel energy calculations (HL). The rate constants of these channels are calculated using the improved canonical variational transition-state theory with the small curvature tunneling correction (ICVT/SCT) method. The calculated results show that, in the whole temperature range, the more favorable reaction channels are Channels 1 and 3. The total ICVT/SCT rate constants of the four channels at the HL//B3LYP/cc-pVTZ level of theory are in good agreement with the available experiment data over the measured temperature ranges, and the corresponding three parameter expression is k(ICVT/SCT) = 3.13 x 10(-20) T(2.70) exp(-11.52/RT) cm3 mole(-1) s(-1) in the temperature range of 250-3000 K. Additionally, the flexibility of the dihedral angle of H2O2 is also discussed to explain the different experimental values. PMID- 16366659 TI - Graph theory for fused cubic clusters of water dodecamer. AB - The stable structures of the fused cubic water cluster (H2O)12 are examined using graph theoretical techniques and ab initio calculations. The calculations are obtained by scanning the symmetry of digraph structures of hydrogen-bond network spanning 12 oxygen atom vertexes. Using the Polya theorem the cycle index expressions for 12 vertexes and 20 edges of a cuboid in point-group symmetry D(4h) are developed. A total of 91 energy-allowed fused cubic structures are obtained, which are classified by 8 point-group symmetries: 1 D(2h), 2 S4, 5 C4, 1 D2, 11 C2, 10 C(i), 1 C(s), and 60 C1. An energy level diagram of the structures reveals 14 bands that correspond to 14 unique two-colored graphs derived from the distributions of four free hydrogens of the cluster. PMID- 16366660 TI - Intramolecular hydrogen bonding and hydrogen atom abstraction in gas-phase aliphatic amine radical cations. AB - The intramolecular hydrogen atom abstraction by the nitrogen atom in isolated aliphatic amine radical cations is examined experimentally and with composite high-level ab initio methods of the G3 family. The magnitude of the enthalpy barriers toward H-atom transfer varies with the shape and size of the cyclic transition state and with the degree of substitution at the nitrogen and carbon atoms involved. The lower barriers are found for 1,5- and 1,6-abstraction, for chairlike transition states, for abstraction reactions in ionized primary amines, and for abstraction of H from tertiary carbon atoms. In most cases, the internal energy required for 1,4-, 1,5-, and 1,6-hydrogen atom abstraction to occur is less than that required for gas-phase fragmentation by simple cleavage of C-C bonds, which explains why H-atom transfer can be reversible and result in extensive H/D exchange prior to the fragmentation of many low-energy deuterium labeled ionized amines. The H-atom transfer to nitrogen is exothermic for primary amine radical cations and endothermic for tertiary amines. It gives rise to a variety of distonic radical cations, and these may undergo further isomerization. The heat of formation of the gauche conformers of the gamma-, delta-, and epsilon distonic isomers is up to 25 kJ mol(-1) lower than that of the corresponding trans forms, which is taken to reflect C-H-N hydrogen bonding between the protonated amino group and the alkyl radical site. PMID- 16366661 TI - Controversy on the first ionization potential of PuO2 (nearly) settled by new experimental evidence. AB - The first ionization potential of the PuO2 molecule was for a long time considered to be 4-5 eV higher than that of UO2. This feature could hardly be explained by the most advanced "ab initio" calculations, which, on the other hand, provide satisfactory results for other actinide oxides. From recent experiments, performed with different techniques, a lower ionization potential of approximately 7 eV was measured, in better agreement with the theoretical predictions. Our recent experiments, where thermally produced ions were measured, make it possible to formulate an accurate relation between the ionization potential of PuO2 and that of PuO: I0(PuO2) = I0(PuO) + 0.42 +/- 0.005 eV. The present uncertainty of I0(PuO) leads to the final assessment, 6.2 < or = I0(PuO2) < or = 6.6 eV, whereby the upper limit is more in line with the aforementioned recent measurements. Considering the still existing uncertainties, one can conclude that these results remove major doubts on the validity of the current theoretical predictions. However, the very small ionization cross section of PuO2 by low-energy electron collisions, which led to the previous spurious assessment of the ion appearance potential, has still an unexplained cause. PMID- 16366663 TI - Comment on "The reaction of acetylene with hydroxyl radicals". PMID- 16366662 TI - 13C NMR relaxation study of molecular motions in tetraphenyltin and tetra(p tolyl)tin in solution. AB - The Woessner approach is applied to the 13C relaxation data for tetraphenyltin (1) and tetra(p-tolyl)tin (2) in CDCl3 solution over the temperature range 5-42 degrees C to obtain correlation times for rotational motions and hence the activation barriers. Quantum mechanical computations were carried out to obtain the rotational energy barriers for comparison. For 2 the relaxation data indicate (1) slower ring rotation than in 1, (2) highly hindered internal rotation of the methyl group. IR and chemical shift data support the hypothesis of hyperconjugation of the methyl correlated with interaction between the pi electrons and the 5d orbitals of tin in the (p-tolyl)Sn moiety to account for the hindrances to the rotations of the ring and the methyl. The activation barrier for the tolyl group rotation is found to be much higher than that for the phenyl rotation. However, the Woessner approach yields an anomalously high barrier for the methyl rotation. An explanation based on correlated rotations of the tolyl ring and the methyl is offered. PMID- 16366664 TI - Monitoring lipase-catalyzed interesterification for bulky fat modification with FT-IR/NIR spectroscopy. AB - This work demonstrates the application of FT-IR and FT-NIR spectroscopy to monitor the enzymatic interesterification process for bulky fat modification. The reaction was conducted between palm stearin and coconut oil (70:30, w/w) with the catalysis of Lipozyme TL IM at 70 degrees C in a batch reactor. The blends and interesterified fat samples in liquid form were measured by attenuated total reflectance based FT-IR (spectra region, 1516-781 cm(-1)) and transmission mode based FT-NIR (spectra region, 5369-4752 cm(-1)) with the temperature of both controlled at 70 degrees C. The samples in solid form were also measured by reflectance-based FT-NIR (spectra regions, 7037-6039 and 5995-5612 cm(-1)) at room temperature. Calibrations of FT-IR and FT-NIR for conversion degrees (evaluated by triglyceride profile), solid fat contents (SFC), and dropping points of interesterified products were carried out by using partial least squares regression. High correlations (r > 0.96) were obtained from cross validations of the data estimated by FT-IR, FT-NIR, and the above-mentioned conventional analytical methods, except for correlations (r = 0.90-0.95) between FT-IR and SFC profiles. Overall, FT-NIR spectroscopy coupled with transmission mode measured at 70 degrees C had the highest correlations, which also had the closest conditions to the sampled products in the process, indicating a great potential for implementation as an on-line control for monitoring the enzymatic interesterification process. PMID- 16366665 TI - Comparison of several methods for the extraction of DNA from potatoes and potato derived products. AB - Eight methods were compared for the extraction of DNA from raw potato tubers, and nine methods were evaluated for the extraction of DNA from dehydrated potato slices, potato flakes, potato flour, potato starch, and two ready-to-eat potato snack foods. Extracts were assessed for yield using a fluorescence-based DNA quantification assay. Real-time amplification of an endogenous gene, sucrose synthase (sus), was used to assess extract and template quality. A CTAB-based method extracted the highest DNA yields from the tuber material. An in-house method, which utilized the Kingfisher magnetic particle processor, yielded the highest template quality from the tubers. For most of the tuber samples, the Kingfisher and CTAB methods recovered the highest levels of amplifiable sus. DNA yields for potato-derived foods generally decreased with the extent that the product had been processed. The methods that utilized the magnetic particle processor delivered the highest template quality from one of the snack products that was particularly high in fat. For most of the remaining processed products, the levels of amplifiable target DNA recovered were roughly correlated with total DNA recovery, indicating that overall yield had greater influence over sus amplification than template quality. The Wizard method was generally the best method for the extraction of DNA from most of the potato-derived foods. PMID- 16366666 TI - A rapid liquid chromatography electrospray ionization mass spectrometry(n) method for evaluation of synephrine in Citrus aurantium L. samples. AB - Immature bitter orange fruit and its extracts have been introduced into the market as an alternative to Ephedra in weight loss products. However, the safety of the immature bitter orange fruit and its extracts is a debated argument due to the presence of synephrine, a constituent known as a sympathomimetic agent. In this paper, we describe the development of a new, rapid, and simple liquid chromatography-electrospray ionization-tandem mass spectrometry method devoted to the quantitative determination of synephrine in bitter orange samples, containing a high quantity of synephrine, and sweet orange samples, known to contain a low level of synephrine but at the same time being one of the main synephrine sources in a normal human diet. Two bitter orange dry extracts containing 5 and 6% sSynephrine and 10 sweet orange samples have been analyzed. Between the sweet orange samples, six were fresh oranges and four were fresh-squeezed juices; in these samples, the synephrine levels ranged from 0.00128 to 0.00349%. PMID- 16366667 TI - Simplified extraction of ginsenosides from American ginseng (Panax quinquefolius L.) for high-performance liquid chromatography-ultraviolet analysis. AB - Four methods were tested for extraction and recovery of six major ginsenosides (Rb1, Rb2, Rc, Rd, Re, and Rg1) found in roots of American ginseng (Panax quinquefolius): method A, sonication in 100% methanol (MeOH) at room temperature (rt); method B, sonication in 70% aqueous MeOH at rt; method C, water extraction (90 degrees C) with gentle agitation; and method D, refluxing (60 degrees C) in 100% MeOH. After 0.5-1 h, the samples were filtered and analyzed by high performance liquid chromatography (HPLC)-UV. A second extraction by methods C and D was done, but 85-90% of ginsenosides were obtained during the first extraction. Lyophilization of extracts did not influence ginsenoside recovery. Method D resulted in the highest significant recoveries of all ginsenosides, except Rg1. Method C was the next most effective method, while method A resulted in the lowest ginsenoside recoveries. Method B led to similar recoveries as method C. All methods used one filtration step, omitted time-consuming cleanup, but maintained clear peak resolution by HPLC, and can be used for quantitative screening of ginsenosides from roots and commercial ginseng preparations. PMID- 16366668 TI - Rapid determination of diphenylamine residues in apples and pears with a single multicommuted fluorometric optosensor. AB - In this work, a single flow injection multicommuted system using solid-surface fluorescence spectroscopy has been explored for the determination of diphenylamine in apples and pears. The native fluorescence signal of diphenylamine retained on the solid support (C18 silica gel) was used for its determination (lambda(exc)/lambda(em) 291/372 nm). The sample treatment consists of a liquid-liquid extraction with acetonitrile followed by a cleanup step using primary-secondary amines. The use of a continuous flow assembly implementing multicommutation, based on a set of three-way solenoid valves controlled by appropriate software, provides the automatic control of sample, carrier, and eluting solution with remarkable advantages in relation to conventional flow injection approaches. Using an optimized sampling time, the proposed method was linear in the range 0.25-5 mg kg(-1) with a detection limit of 0.06 mg kg(-1) and RSD (percent) values better than 3%. The proposed method was successfully applied to the analysis of diphenylamine in different apple and pear samples fortified at different concentrations, and recoveries between 78 and 104% were found. The results obtained illustrate the usefulness of the proposed method for the screening and evaluation of postharvest treatment of crops possibly containing diphenylamine. PMID- 16366669 TI - Liquid chromatography-electrospray ionization-tandem mass spectrometry method for the determination of epoxidized soybean oil in food products. AB - Epoxidized soybean oil (ESBO) is widely used as a plasticizer and stabilizer in such polymers [poly(vinyl chloride) in particular] commonly adopted for manufacturing of gaskets of the lids for glass jars and plastic films for food packaging. Human exposure to ESBO and its derivatives is likely to occur over a lifetime with a significant variation according to life stage. A reversed phase liquid chromatography interfaced with electrospray ion trap tandem mass spectrometry method for the determination of ESBO in foods was developed. A simple sample treatment procedure entailing the use of an extraction step with dichloromethane without any further cleanup was proved. Chromatographic separation was performed using two C18 columns with an aqueous acetic acid acetone-acetonitrile mixture as the mobile phase under gradient conditions. The method was validated in terms of detection limits (4 mg kg(-1)), quantitation limits, linearity (established over 2 orders of magnitude), recovery (good mean recoveries, higher than 90% for all of the signals detected), precision (RSD% < 8), and trueness. The applicability of the method to the determination of ESBO in different food matrices (in particular those rich in edible oil) was demonstrated, and the performances were compared to those reachable by the commonly well-known gas chromatography-mass spectrometry procedure. PMID- 16366670 TI - Analysis of tea catechins in human plasma by high-performance liquid chromatography with solid-phase extraction. AB - Accurate monitoring of tea catechins in biological samples might provide a means of better evaluation of their benefits. The aim of the present study was to develop a rapid method for extracting tea catechins from human plasma samples with a solid-phase extraction technique and to subsequently measure their concentrations using an HPLC system. A human plasma sample spiked with known concentrations of the analyte standards was passed through a Waters Oasis HLB cartridge. After repeated washing, tea catechins were eluted with 70% dimethylformamide containing 0.1% phosphoric acid, and the resulting eluate was injected into an HPLC system. Analytes were separated on a reverse-phase C18 column using an isocratic mobile phase and detected electrochemically. The coefficient of variation for inter- and intraday reproducibility was less than 5.0% and 6.4%, respectively. Linearity was established for the concentration range of 0.01-1.0 microM. The method was successfully applied to measure tea catechin concentrations in the plasma of two healthy subjects who received a single ingestion of a green tea beverage. The proposed method enables the rapid and accurate quantitation of plasma tea catechins and might prove useful for the evaluation of beneficial health effects of tea consumption. PMID- 16366671 TI - Isolation of 4-methylthio-3-butenyl glucosinolate from Raphanus sativus sprouts (kaiware daikon) and its redox properties. AB - The most promising among glucosinolates (GLs) are those bearing in their aglycon an extra sulfur function, such as glucoraphasatin (4-methylthio-3-butenyl GL; GRH) and glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE). The GRE/GRH redox couple is typically met among secondary metabolites of Raphanus sativus L. and, whereas GRE prevails in seeds, GRH is the major GL in full-grown roots. During the 10 days of sprouting of R. sativus seeds, the GRE and GRH contents were determined according to the Eurpean Union official method (ISO 9167-1). In comparison to the seeds, the GRE content in sprouts decreased from about 90 to about 12 micromol g(-1) of dry weight (dw), whereas a 25-fold increase--from about 3 to 76 micromol g(-1) of dw--of the GRH content was measured. An efficient pure GRH gram-scale production process from R. sativus (kaiware daikon) sprouts resulted in significant yield improvement of up to 2.2% (dw basis). The reaction of GRH with both H2O2 and ABTS*+ radical cation was investigated. Whereas H2O2 oxidation of GRH readily resulted in complete transformation into GRE, ABTS*+ caused complete decay of the GL. Even though not directly related to its radical scavenging activity, the assessed reducing capacity of GRH suggests that R. sativus sprouts might possess potential for health benefits. PMID- 16366672 TI - Metabolism of the olive oil phenols hydroxytyrosol, tyrosol, and hydroxytyrosyl acetate by human hepatoma HepG2 cells. AB - To study the potential hepatic metabolism of olive oil phenols, human hepatoma HepG2 cells were incubated for 2 and 18 h with hydroxytyrosol, tyrosol, and hydroxytyrosyl acetate, three phenolic constituents of olive oil. After incubation, culture media and cell lysates were hydrolyzed with beta glucuronidase and sulfatase and analyzed by LC-MS. In vitro methylation, glucuronidation, and sulfation of pure phenols were also performed. Methylated and glucuronidated forms of hydroxytyrosol were detected at 18 h of incubation, together with methylglucuronidated metabolites. Hydroxytyrosyl acetate was largely converted into free hydroxytyrosol and subsequently metabolized, yet small amounts of glucuronidated hydroxytyrosyl acetate were detected. Tyrosol was poorly metabolized, with <10% of the phenol glucuronidated after 18 h. Minor amounts of free or conjugated phenols were detected in cell lysates. No sulfated metabolites were found. In conclusion, olive oil phenols can be metabolized by the liver as suggested by the results obtained using HepG2 cells as a hepatic model system. PMID- 16366673 TI - Effects on the flavor and oxidative stability of stripped soybean and sunflower oils with added pure tocopherols. AB - Effects of tocopherols on the oxidative stability of stripped vegetable oils were studied by adding pure tocopherols--alpha, beta, gamma, and delta--in their naturally occurring proportions in soybean and sunflower oils to the triacylglycerols (TAG) of soybean and sunflower oils. Soybean and sunflower oils were purified by stripping all minor constituents, leaving the triacylglycerols. Pure tocopherols in the proportion typical of sunflower oil--high alpha, low gamma, and low delta--were added to purified sunflower oil and to purified soybean oil. Pure tocopherols in the proportion typical of soybean oil--low alpha, high gamma, and high delta--were added to the purified oils. Oils were subjected to accelerated autoxidation using oven storage at 60 degrees C in the dark and accelerated photooxidation at 7500 lx light intensity at 30 degrees C. Oxidation levels of aged oils were measured by the formation of both peroxides and volatile compounds and by flavor analysis. Results from substituting the tocopherol profile from one oil type to another varied on the basis of whether they were oxidized in the dark or in the light. For example, during autoxidation in the dark, soybean oil with the typical soybean tocopherol profile had the lowest levels of peroxides and total volatile compounds, whereas sunflower oil with the sunflower tocopherol profile had the highest levels. In flavor analyses of the same oils, sunflower oil with the soybean tocopherol profile was the most stable. Soybean oil with the profile of sunflower tocopherols was the least stable in dark oxidation. In contrast to the data from autoxidation in the dark, addition of tocopherols typical of sunflower oil significantly improved light stability of both oil types compared to the addition of soybean tocopherols to sunflower oil. The tocopherol profile typical of soybean oil was significantly more effective in inhibiting autoxidation in the dark; however, the tocopherol profile typical of sunflower oil inhibited light oxidation significantly more than the soybean tocopherol profile. PMID- 16366674 TI - Investigation of Australian olive mill waste for recovery of biophenols. AB - Olive mill waste is a potential source for the recovery of phytochemicals with a wide array of biological activities. Phytochemical screening of hexane, methanol, and water extracts revealed a diversity of compounds, perhaps overlooked in previous studies through intensive cleanup procedures. Methanol and water extracts contained large amounts of biophenols, and further testing of polar extraction solvents, including ethyl acetate, ethanol, propanol, acetone, acetonitrile, and water/methanol mixtures, highlighted the latter as the solvent of choice for extraction of the widest array of phenolic compounds. Stabilization of the resulting extract was best achieved by addition of 2% (w/w) sodium metabisulfite. Quantitative data are reported for nine biophenols extracted using 60% (v/v) methanol in water with 2% (w/w) sodium metabisulfite. Six compounds had recoveries of greater than 1 g/kg of freeze-dried waste: hydroxytyrosol glucoside, hydroxytyrosol, tyrosol, verbascoside, and a derivative of oleuropein. PMID- 16366675 TI - Sweet and sour cherry phenolics and their protective effects on neuronal cells. AB - The identification of phenolics from various cultivars of fresh sweet and sour cherries and their protective effects on neuronal cells were comparatively evaluated in this study. Phenolics in cherries of four sweet and four sour cultivars were extracted and analyzed for total phenolics, total anthocyanins, and their antineurodegenerative activities. Total phenolics in sweet and sour cherries per 100 g ranged from 92.1 to 146.8 and from 146.1 to 312.4 mg gallic acid equivalents, respectively. Total anthocyanins of sweet and sour cherries ranged from 30.2 to 76.6 and from 49.1 to 109.2 mg cyanidin 3-glucoside equivalents, respectively. High-performance liquid chromatography (HPLC) analysis revealed that anthocyanins such as cyanidin and peonidin derivatives were prevalent phenolics. Hydroxycinnamic acids consisted of neochlorogenic acid, chlorogenic acid, and p-coumaric acid derivatives. Glycosides of quercetin, kaempferol, and isorhamnetin were also found. Generally, sour cherries had higher concentrations of total phenolics than sweet cherries, due to a higher concentration of anthocyanins and hydroxycinnamic acids. A positive linear correlation (r2 = 0.985) was revealed between the total anthocyanins measured by summation of individual peaks from HPLC analysis and the total anthocyanins measured by the pH differential method, indicating that there was in a close agreement with two quantifying methods for measuring anthocyanin contents. Cherry phenolics protected neuronal cells (PC 12) from cell-damaging oxidative stress in a dose-dependent manner mainly due to anthocyanins. Overall results showed that cherries are rich in phenolics, especially in anthocyanins, with a strong antineurodegenerative activity and that they can serve as a good source of biofunctional phytochemicals in our diet. PMID- 16366676 TI - Absorption and bioavailability of artepillin C in rats after oral administration. AB - Artepillin C (AC), an active ingredient of Brazilian propolis, permeates intact across Caco-2 cells by transcellular passive diffusion. The permeation of AC across Caco-2 cells is as efficient as that of phenolic acids and the microbial metabolites of poorly absorbed polyphenols, which are actively absorbed by the monocarboxylic acid transporter (MCT) (Biochim. Biophys. Acta 2005, 1713, 138 144). Here, the absorption of orally administered AC in rats has been studied to evaluate its pharmacokinetics and bioavailability in vivo in comparison with those of p-coumaric acid (CA), a substrate of MCT. Rats were given 100 micromol/kg of body weight of AC or CA, and blood was subsequently collected from the portal vein and abdominal artery. AC, CA, and their metabolites were quantified by coulometric detection using HPLC-ECD. The serum concentration of intact AC and CA in the portal vein peaked at 5-10 min after administration, with a C(max) of 19.7 micromol/L for AC and 74.8 micromol/L for CA. The area under the curve (AUC) for intact AC and CA in the portal vein was calculated from the serum concentration as 182.6 and 3057.3 micromol.min.L(-1), respectively. The absorption efficiency of CA was about 17-fold higher than that of AC. Furthermore, the bioavailability of CA was about 278-fold higher than that of AC, and the ratio of AUC in the abdominal artery to AUC in the portal vein was 0.04 and 0.70, for AC and CA, respectively. Thus, AC is likely to be more susceptible to hepatic elimination than is CA. The bioactive compound of AC in vivo should be investigated further. PMID- 16366677 TI - Natural compounds derived from foods modulate nitric oxide production and oxidative status in epithelial lung cells. AB - The effects of natural antioxidants on nitric oxide (NO) modulation and oxidative status were determined in rat epithelial lung cells (L-2). Cells were stimulated with cytokines and treated with one of the following: resveratrol, soybean saponin group B (SSB), quercetin, genistein, olive leaf polyphenol concentrate (OLPC), or N-acetyl-L-cystein (NAC). NAC had no effect on NO levels, whereas resveratrol and OLPC were found to be effective in reducing nitrite levels, modifying iNOS mRNA, and decreasing free radical production. OLPC affected the levels of MnSOD while resveratrol did not, indicating that they act via different pathways. Quercetin and genistein reduced nitrite levels without affecting iNOS levels, presumably by scavenging NO. SSB did not affect nitrite levels, but exposure did reduce iNOS mRNA expression and protein levels, possibly due to antioxidant activity. Naturally occurring antioxidants, in particular resveratrol and OLPC, may have therapeutic potential in the treatment of inflammatory diseases. PMID- 16366678 TI - Phenolic antioxidants (hydrolyzable tannins, flavonols, and anthocyanins) identified by LC-ESI-MS and MALDI-QIT-TOF MS from Rosa chinensis flowers. AB - Rosa chinensis (Yuejihua) is a well-known ornamental plant, and its flowers are commonly used in traditional Chinese medicine. Methanolic crude extracts of dried R. chinensis flowers were used for simultaneous determination of phenolic constituents by liquid chromatography-mass spectrometry (LC-MS) and matrix assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS). A total of 36 known and unknown phenolics were identified as hydrolyzable tannins, flavonols, and anthocyanins, mainly including gallotannins (mono-, di-, or trigalloylglucopyranosides), ellagitannins, quercetin, quercetin/kaempferol mono- and diglycosides, and cyanidin/pelargonidin diglycosides. MALDI-QIT-TOF MS was applied not only to verify most phenolics isolated and identified by LC-MS but also to tentatively identify two ellagitannins (rugosins B and C) not isolated and unidentified by LC-MS. This study is the first to demonstrate the rapid and successful use of MALDI-QIT-TOF MS and LC-MS to directly and simultaneously identify phenolics in the crude extracts of R. chinensis flowers without any purification. The antioxidant activity of the crude extracts from R. chinensis flowers was also measured with three assay methods. The results showed that the phenolic antioxidants from R. chinensis flowers exhibited very strong radical scavenging effect and antioxidant power. High levels of flavonols and hydrolyzable tannins might be important bioactive principles in the dried R. chinensis flowers. PMID- 16366679 TI - Nicotine carboxylate insecticide emulsions: effect of the fatty acid chain length. AB - The effect of fatty acid chain length on nicotine carboxylate insecticide emulsions has been studied in terms of particle size, interfacial tension, nicotine encapsulation on emulsion droplets, and bioactivity. The particle size of the nicotine emulsion and the interfacial tension at the nicotine carboxylate oil phase (0.03 M)--Tween 80 aqueous phase (0.001 M) were affected in a similar way by the change in the fatty acid chain length, which was correlated by the packing conformation of Tween 80 and nicotine carboxylate molecules as obtained by AM1 theoretical calculations. The amount of encapsulated nicotine inside the nicotine carboxylate emulsion droplets influenced the insecticide bioactivity of nicotine; this relationship was explained in terms of the acid value of the different fatty acids used to prepare the nicotine formulation. PMID- 16366680 TI - Triterpenoid glycosides from leaves of Medicago arborea L. AB - Eighteen triterpene saponins (1-18) from Medicago arborea leaves have been isolated and their structures elucidated by spectroscopic, spectrometric (1D and 2D NMR, FAB-MS, ESI-MS/MS), and chemical methods. They have been identified as glycosides of medicagenic, zanhic, and 2beta-hydroxyoleanolic acids, soyasapogenol B, bayogenin, and 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid. Twelve of them, identified as 3-O-beta-D-glucopyranosyl-28-O-[alpha-L arabinopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid (3), 3-O-beta-D-glucopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4) [alpha-L-arabinopyranosyl-(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranoside] zanhic acid (4), 3-O-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranosyl(1-->2)-beta-D-glucopyranosyl]-2beta-hydroxyoleanolic acid (5), 3 O-beta-D-glucuronopyranosyl-28-O-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranoside]medicagenic acid (6), 3-O-beta-D-glucuronopyranosyl-28-O-[beta D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranoside]bayogenin (9), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside] 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid (10), 3-O-beta-D glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1- >3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (12), 3 O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L arabinopyranoside(1-->3)]-alpha-L-rhamnopyrano-syl(1-->2)-alpha-L arabinopyranoside]zanhic acid (13), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D xylopyrano-syl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranoside]zanhic acid (14), 3-O-[alpha-L-arabinopyranosyl-(1-->2)-beta-D glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4) [beta-D-apiofurano-syl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranoside]zanhic acid (16), 3-O-[beta-D-glucopyrano-syl(1-->2)-beta-D glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranosyl(1- >3)]-alpha-L-rhamno-pyranosyl (1-->2)-alpha-L-arabinopyranoside]zanhic acid (17), and 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyrano side]medicagenic acid (18), are reported as new natural compounds. The presence of the aldehydic group on the sapogenin moiety of saponin 10 is discussed in the framework of a possible elucidation of the biosynthesis of these metabolites. PMID- 16366681 TI - Heterologous expression of type I antifreeze peptide GS-5 in baker's yeast increases freeze tolerance and provides enhanced gas production in frozen dough. AB - The demand for frozen-dough products has increased notably in the baking industry. Nowadays, no appropriate industrial baker's yeast with optimal gassing capacity in frozen dough is, however, available, and it is unlikely that classical breeding programs could provide significant improvements of this trait. Antifreeze proteins, found in diverse organisms, display the ability to inhibit the growth of ice, allowing them to survive at temperatures below 0 degrees C. In this study a recombinant antifreeze peptide GS-5 was expressed from the polar fish grubby sculpin (Myoxocephalus aenaeus) in laboratory and industrial baker's yeast strains of Saccharomyces cerevisiae. Production of the recombinant protein increased freezing tolerance in both strains tested. Furthermore, expression of the GS-5 encoding gene enhanced notably the gassing rate and total gas production in frozen and frozen sweet doughs. These effects are unlikely to be due to reduced osmotic damage during freezing/thawing, because recombinant cells showed growth behavior similar to that of the parent under hypermosmotic stress conditions. PMID- 16366682 TI - Distortion of genetically modified organism quantification in processed foods: influence of particle size compositions and heat-induced DNA degradation. AB - Milling fractions from conventional and transgenic corn were prepared at laboratory scale and used to study the influence of sample composition and heat induced DNA degradation on the relative quantification of genetically modified organisms (GMO) in food products. Particle size distributions of the obtained fractions (coarse grits, regular grits, meal, and flour) were characterized using a laser diffraction system. The application of two DNA isolation protocols revealed a strong correlation between the degree of comminution of the milling fractions and the DNA yield in the extracts. Mixtures of milling fractions from conventional and transgenic material (1%) were prepared and analyzed via real time polymerase chain reaction. Accurate quantification of the adjusted GMO content was only possible in mixtures containing conventional and transgenic material in the form of analogous milling fractions, whereas mixtures of fractions exhibiting different particle size distributions delivered significantly over- and underestimated GMO contents depending on their compositions. The process of heat-induced nucleic acid degradation was followed by applying two established quantitative assays showing differences between the lengths of the recombinant and reference target sequences (A, deltal(A) = -25 bp; B, deltal(B) = +16 bp; values related to the amplicon length of the reference gene). Data obtained by the application of method A resulted in underestimated recoveries of GMO contents in the samples of heat-treated products, reflecting the favored degradation of the longer target sequence used for the detection of the transgene. In contrast, data yielded by the application of method B resulted in increasingly overestimated recoveries of GMO contents. The results show how commonly used food technological processes may lead to distortions in the results of quantitative GMO analyses. PMID- 16366683 TI - Biochemical study of leaf browning in minimally processed leaves of lettuce (Lactuca sativa L. var. acephala). AB - A series of biochemical parameters, including the concentration of total ascorbic acid (ASA(tot)) and the activities of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidases (PODs), was investigated during cold storage (72 h at 4 degrees C in the dark) in fresh-cut (minimally processed) leaves of two lettuce (Lactuca sativa L. var. acephala) cultivars differing in the susceptibility to tissue browning: Green Salade Bowl (GSB), susceptible, and Red Salade Bowl (RSB), resistant. The two cultivars showed differences also at the biochemical level. The content in ASA(tot) increased in RSB, as a consequence of increased DHA concentration; conversely, ASA(tot) diminished in GSB, in which ASA was not detectable after 72 h of storage, thus suggesting a disappearance of ascorbate (both ASA and DHA) into nonactive forms. The antioxidant capacity (as determined by using FRAP analysis) decreased significantly during storage in RSB, while a strong increase was observed in GSB. PAL activity increased soon after processing reaching a maximum by 3 h, then it declined to a relatively constant value in RSB, while in GSB it showed a tendency to decrease in the first few hours from harvest and processing. POD activity, at least for chlorogenic acid, increased significantly during storage only in GSB. PMID- 16366684 TI - Application of fluorescence spectroscopy in the evaluation of light-induced oxidation in cheese. AB - Light-induced oxidation of semihard cheese has been evaluated by fluorescence spectroscopy. The cheese was packaged in two packaging materials and exposed to different storage conditions, which included light/dark storage, oxygen availability, and storage time (0, 4, 7, 14, 21, 42, 70, or 84 days). Fluorescence excitation-emission matrices (EEM) were analyzed by PARAFAC, which gave an estimation of the pure excitation and emission spectra of the fluorophores and the concentrations of these. This analysis showed the presence of components such as tryptophan, tyrosine, vitamin A, fluorescent oxidation products, and riboflavin. Effects of packaging material, light or dark storage, and storage time were seen. However, there was no effect of the oxygen availability on the fluorescence measurements. The score values obtained by the PARAFAC models and chemical and physical measurements were analyzed together by principal component analysis (PCA). The loadings showed a separation of the variables into three groups; the first group was related to oxidation, the second group was related to the degradation of both riboflavin and vitamin A, and the third group was linked to the protein structure. PMID- 16366685 TI - Isomeric influence on the oxidative coloration of phenolic compounds in a model white wine: comparison of (+)-catechin and (-)-epicatechin. AB - The reactions of (+)-catechin and (-)-epicatechin with glyoxylic acid were studied in a model white wine solution. When the reactions were performed in darkness at 45 degrees C, the (-)-epicatechin concentration decreased more rapidly than that of (+)-catechin, and the (-)-epicatechin sample had twice the 440 nm absorbance of the (+)-catechin sample after the 14 day incubation period. The main pigments generated were identified as xanthylium cation pigments regardless of the isomeric character of the phenolic compound. Using a combination of absorbance and ion current data, the xanthylium cation pigments generated from (-)-epicatechin were found to have combined molar absorptivity coefficients 1.8 times that of the xanthylium cation pigments generated from (+) catechin. The implication of these results on the development of an index of white wine oxidation susceptibility is discussed. PMID- 16366686 TI - Kinetics of acrylamide formation and elimination during heating of an asparagine sugar model system. AB - The kinetics of acrylamide (AA) was analyzed by heating a simple model system consisting of asparagine and glucose, fructose, or sucrose (0.01 M, pH 6) at temperatures between 140 and 200 degrees C. The AA concentration appeared to be the net result of simultaneous formation and elimination. A general kinetic model describing the AA yield was identified, and kinetic parameters were obtained by nonlinear regression on the nonisothermally derived data. On the basis of kinetic parameters, the AA formation appeared to proceed faster and to be more temperature sensitive in the asparagine-glucose than in the asparagine-fructose model system. The AA elimination kinetics, on the other hand, was similar. Significantly less AA was formed in the asparagine-sucrose model system as compared to the model systems with glucose or fructose. PMID- 16366687 TI - Effect of screwcap and cork closures on SO2 levels and aromas in a Sauvignon Blanc wine. AB - The development of a Sauvignon Blanc wine sealed under screwcap and cork was undertaken using different fill heights and initial levels of free SO2 (20, 25, and 30 mg/L) over 2 years. More SO2 was lost for wines under cork over the first 3 months, corresponding to a higher level of dissolved oxygen at bottling. From this time wines under cork and screwcap lost SO2 at a similar rate and retained dissolved CO2 equally well, indicating that both types of closure presented a similar effective barrier to gas movement. After 2 years in the bottle, the different treatments retained similar levels of the volatile thiols 3 mercaptohexyl acetate (3MHA) and 3-mercaptohexanol (3MH) responsible for fruity aromas, with initial SO2 levels having no effect, but the thiol concentrations were 18-23% lower under cork, which may be due to absorption of volatiles into the cork. Levels of polyphenols such as caftaric acid and the absorbance at 420 nm were the same for wines under cork and screwcap, whereas some indication was given that more oxidation occurred with a lower level of initial free SO2. Although the different treatments were not readily distinguished by a sensory panel, the data for individual wines showed a positive correlation between passion fruit descriptors and levels of 3MHA and 3MH. PMID- 16366688 TI - Separation of muscle membrane from alkali-solubilized fish muscle proteins. AB - Treatment with Ca2+ and citric acid improved membrane removal from muscle homogenates solubilized at pH 10.5 by centrifugation at 4000 g for 15 min. The percentage of phospholipid removed from muscle homogenates increased with increasing Ca2+ concentrations at 1 mM citric acid. More than 85% phospholipid and 45% protein in the muscle homogenates were removed at Ca2+ concentrations of >20 mM in the presence of 1 mM citric acid. At 8 mM Ca2+, addition of citric acid at 5 mM improved phospholipid removal to approximately 78% from 58% in its absence. Because treatment with 8 mM Ca2+ alone can remove significant amounts of phospholipid, it is likely that Ca2+ played the major role in membrane removal in muscle homogenates solubilized at pH 10.5. PMID- 16366689 TI - Anhydrous goat's milk fat: thermal and structural behavior. 1. Crystalline forms obtained by slow cooling. AB - The thermal and structural behaviors of anhydrous goat's milk fat (AGMF) have been determined as a function of temperature using a powerful technique allowing simultaneous time-resolved synchrotron X-ray diffraction as a function of temperature (XRDT) and high-sensivity differential scanning calorimetry (DSC) measurements from the same sample. This first paper, aiming at the characterization of the physical properties of AGMF, we examine crystalline organizations made by triacylglycerols (TG) upon slow cooling at /dT/dt/ = 0.1 degrees C/min from 45 to -20 degrees C in order to approach system equilibrium. Three overlapped exotherms were observed by DSC upon cooling, whereas four endotherms were found on the subsequent heating at 1 degrees C/min. XRDT evidenced that AGMF crystallizes under four different lamellar structures, two with double-chain length packings at 41.5 and 38.2 angstroms and two with triple chain lengths of 72 and 64.7 angstroms stacking. Simultaneous wide-angle XRDT has shown that initial nucleation mainly occurs in a packing of beta' type from approximately 26 degrees C, although some transient presence of alpha was detected. The absence of polymorphic transition, on heating, until final melting (approximately 40 degrees C) demonstrated the relative stability of the structures formed. PMID- 16366690 TI - Isolation and enzymatic digestion of body complex of soybean seed. AB - The body complex of the soybean seed (BCSS) was isolated from the single cells (27.2%) by a sequential procedure of autoclaving with water, cellulase digestion for the primary cell wall, pectinase digestion for the secondary cell wall, and defatting with hexane washing. Its characteristics were then investigated. The defatted BCSS (DBCSS) consisted of protein (76.5%) and mannose-rich carbohydrates (3.2%). Screening of the food-processing protease for the digestion of DBCSS was carried out, and a kind of alkaline protease was selected. The inner protein of DBCSS was easily extracted with 0.1 M sodium carbonate buffer, pH 10, and the insoluble shell of the body complex (SDBCSS) was left. SDBCSS consisted of hydrophobic amino acid-rich protein. SDBCSS was easily digested by the selected alkaline protease. SDBCSS was dissolved by boiling with sodium dodecyl sulfate mercaptoethanol, and it was found to consist of a protein of approximately 3 kDa. The high enzymatic digestion including the selected protease for soybean seed and defatted soybean meal was carried out; both were extracted and digested with a yield of >99.5%. The final indigestible residue was found as paired hexagonal and filamentous organs of the soybean cells. PMID- 16366691 TI - Carotenoid, chlorophyll, and chlorophyll-derived compounds in grapes and port wines. AB - Carotenoids and chlorophyll-derived compounds in grapes and Port wines were investigated by HPLC-DAD and HPLC-DAD-MS (ESP+) analysis. A total of 13 carotenoid and chlorophyll-derived compounds are formally reported in grapes, 3 are identified for the first time, pheophytins a and b and (13Z)-beta-carotene, and 3 others remain unknown. In Port wines 19 compounds with carotenoid or chlorophyll-like structures are present, 8 still unidentified. The young wines showed higher total carotenoid content and chlorophyll-like compounds compared to aged Ports, with lutein and beta-carotene as major carotenoids. Among samples analyzed of monovarietal Vitis vinifera L. cultivar wines produced with the five most important Douro varieties, Tinta Roriz contained the highest levels of carotenoids and Touriga Franca the lowest. The forced-aging study indicated that lutein was more sensitive to temperature than beta-carotene. Additionally, aged wines showed higher ratios of beta-carotene/lutein concentrations compared to new Ports. Rates of degradation of chlorophyll derivative compounds were higher than those for carotene and lutein. PMID- 16366692 TI - Protection of originality of Tokaji Aszu: amines and organic acids in botrytized wines by high-performance liquid chromatography. AB - Amine and organic acid composition of Aszu wines from the Tokaj region of Hungary, nonbotrytized Hungarian wines from different regions, and foreign botrytized wines were analyzed by high-performance liquid chromatography. Hungarian and foreign wines (36 Hungarian and seven foreign botrytized wines) were compared in different ways by calculation of ratios of given amine compounds, analyses of variance, principal component, and discriminant analysis. In wines, putrescine and in some samples 3-methyl-butylamine and/or phenyl ethylamine were found in remarkable concentrations, while in botrytized wines four other amines were verified in high concentration. Good separation between Aszu and foreign botrytized wines was found by calculation of the amine component's ratio. The first two principal components of the principal component analysis accounted for 77 and 84% of the total variance in the data of amines and acids, respectively. The component scores of samples grouped according to Aszu, foreign botrytized, and nonbotrytized wines. Linear discriminant analysis was used for differentiation of Aszu, foreign botrytized, and normal wines. Using nine amines and two acids as variables, the correct classification was 97.6%. On the basis of results, an objective evaluation method can be elaborated for quality control in order to protect the authenticity and origin of wine specialties made from botrytized grapes. PMID- 16366693 TI - Sensory and foaming properties of sparkling cider. AB - The effect of yeast strain and aging time on the chemical composition, analytical, and sensory foam properties of sparkling ciders has been studied. The analytical foam parameters (foamability, HM; Bikerman coefficient, sigma; and foam stability time, T(s)) were significantly influenced by aging and yeast strain. The sensory attributes (initial foam, foam area persistence, bubble size, foam collar, and overall foam quality) improved with aging time. Likewise, the yeast strain positively influenced the assessment of initial foam, foam area persistence, number of bubble chains, and overall foam quality. Significant and positive correlations were found between alcoholic proof, dry extract, total and volatile acidities, total phenols and total proteins, and sigma, whereas HM was negatively correlated with specific gravity, alcoholic proof, dry extract, and total proteins. PMID- 16366694 TI - Flavone C-glycosides from leaves of Oxalis triangularis. AB - The flavone C-glycosides luteolin 6-C-(2''-O-beta-xylopyranosyl-beta glucopyranoside) (1), apigenin 6-C-(2''-O-alpha-rhamnopyranosyl-beta glucopyranoside) (2), apigenin 6-C-(2''-O-beta-xylopyranosyl-beta glucopyranoside) (3), apigenin 6-C-(2''-O-(6'''-(E)-caffeoylglucoside)-beta glucopyranoside) (4), and apigenin 6-C-(2''-O-(6'''-(E)-p-coumaroylglucoside) beta-glucopyranoside) (5) have been isolated from the purple leaves of Oxalis triangularis. Compound 4 is new, while 5 has previously been isolated from Cucumis sativus after treatment with silicon and infection with Sphaerotheca fuliginea. Signal duplication in the NMR spectra of 2, 4, and 5 revealed the presence of rotameric conformers, created by rotational hindrance at the C(sp3) C(sp2) glucosyl-flavone linkage in these flavone C-glycosides. PMID- 16366695 TI - Natural variability of metabolites in maize grain: differences due to genetic background. AB - Understanding the impact of genetic diversity on crop biochemical composition is a prerequisite to the interpretation and potential relevance of biochemical differences experimentally observed between genotypes. This is particularly important in the context of comparative safety assessments for crops developed by new technologies such as genetic engineering. To interrogate the natural variability of biochemical composition, grain from seven maize hybrids grown at four geographically distinct sites in Europe was analyzed for levels of proximates (fat, protein, moisture, ash, and carbohydrates), fiber, amino acids, fatty acids, four vitamins, nine minerals, and secondary metabolites. Statistical evaluation of the compositional data at the p < 0.05 level compared each hybrid against every other hybrid (head-to-head) for all analytes at each site and then across all sites to understand the factors contributing to variability. Of the 4935 statistical comparisons made in this study, 40% (1986) were found to be significant. The magnitude of differences observed, as a percent, ranged between 0.84 and 149% when all individual sites and the combined sites were considered. The large number of statistically significant differences in the levels of these analytes between seven commercial hybrids emphasizes the importance of genetic background and environment as determinants of the biochemical composition of maize grain, reflects the inherent natural variability in those analytes across a representative sampling of maize hybrids, and provides a baseline of the natural range of these nutritional and antinutritional components in maize for comparative compositional assessments. PMID- 16366696 TI - Fatty acid composition of traditional and novel forages. AB - Managing the fatty acid composition of grazing ruminant diets could lead to meat and milk products that have higher conjugated linoleic acid (CLA) concentrations, but forage fatty acid dynamics must be more fully understood for a range of forages before grazing systems can be specified. The fatty acid profiles of 13 different forages, including grasses, legumes, and forbs, grown under greenhouse conditions, were determined. Three separate harvests, at 3-week intervals, were made of each plant material. alpha-Linolenic [C18:3, 7.0-38.4 mg g(-1) of dry matter (DM)], linoleic (C18:2, 2.0-10.3 mg g(-1) of DM), and palmitic (C16:0, 2.6 7.5 mg g(-1) of DM) acids were the most abundant fatty acids in all species at each harvest, together representing approximately 93% of the fatty acids present. Concentrations of fatty acids declined as plants developed, but the fractional contribution of each fatty acid to total fatty acids remained relatively stable over time. Grasses had a uniform composition across species with a mean of 66% of total fatty acids provided by C18:3, 13% by C18:2, and 14% by C16:0. The fractional contribution of C18:3 to total fatty acids was lower and more variable in forbs than in grasses. Intake of fatty acid by grazing ruminants would be affected by the forage species consumed. PMID- 16366697 TI - Assessment of antioxidant activity of cane brown sugars by ABTS and DPPH radical scavenging assays: determination of their polyphenolic and volatile constituents. AB - Seven cane brown sugars (four from La Reunion, two from Mauritius, and one from France) were investigated for their polyphenol content and volatile composition in relation to their free radical scavenging capacity determined by ABTS and DPPH assays. The thin layer coated on the sugar crystal was extracted by Soxhlet extractor with dichloromethane. The volatile compounds of brown sugars were studied by GC-MS, and 43 compounds were identified. The total phenolic content of brown sugars was determined according to the Folin-Ciocalteu method. Phenolic compounds were quantified in the brown sugar extracts by LC-UV-ESI-MS. Brown sugar aqueous solutions exhibited weak free radical scavenging activity in the DPPH assay and higher antioxidant activity in the ABTS assay at relatively high concentration. The brown sugar extracts showed interesting free radical scavenging properties despite the low concentration of phenolic and volatile compounds. Sugar is a common foodstuff traditionally used for its sweetening properties, which might be accompanied by antioxidant properties arising from molecules (polyphenols, Maillard products) other than sucrose of the cane brown sugars. PMID- 16366698 TI - In vitro sensitivity of Botrytis cinerea to anthraquinone and anthrahydroquinone derivatives. AB - The effect on mycelial growth of the fungus Botrytis cinerea of a set of structurally related tricyclic hydroquinones [9,10-dihydroxy-4,4-dimethyl-2,3,5,8 tetrahydroantracen-1(4H)-one and 9,10-dihydroxy-4,4-dimethyl-5,8-dihydroanthracen 1(4H)-one derivatives] and tricyclic quinones [4,4-dimethylanthracen-1,9,10(4H) trione derivatives] was studied. In general, the anthraquinones presented higher activity than the anthrahydroquinones. Anthraquinone and anthrahydroquinone derivatives with methyl groups on the A ring showed higher antifungal activity than the unsubstituted ones, 4,4,6,7-tetramethyl-(4H)-anthracene-1,9,10-trione being the most active compound of this set. The presence of a polar group such as hydroxymethyl reduced the activity. The effect of two anthrahydroquinones and two anthraquinones on the conidia germination of the fungus was also determined. Anthrahydroquinones did not affect the germination. The most active compound was 4,4-dimethylanthracene-1,9,10(4H)-trione, with 100% inhibition of germination at 7 h of incubation. These results again suggest that the structure of the anthraquinones is important in exerting an antifungal effect on B. cinerea. Furthermore, possible mechanisms of action of compound 4,4-dimethylanthracene 1,9,10(4H)-trione were studied. This compound did not produce lipoperoxidation of membrane and did not induce the formation of oxygen reactive species, but it was able to permeabilize the plasmatic membrane of B. cinerea, increasing the phosphorus concentration in the intracellular medium. PMID- 16366699 TI - Evaluation of residues of essential oil components in honey after different anti varroa treatments. AB - Apiary trials on the use of three different treatments (Apilife Var, thymol solution in olive oil, and thymol solution in ethanol) for the control of Varroa destructor were conducted in Aragon (northeastern Spain). For the evaluation of the presence of residues of these treatments in honey an analytical method was developed. The method is applied to analyze honey samples before and after treatments with the acaricides mentioned. A solid-phase extraction on trifunctional silane SPE C18 cartridge and gas chromatography separation using a flame ionization detector allow reliable and precise determination of residues of thymol, menthol, eucalyptol, and camphor in honey. The results indicate that camphor is present in only low concentrations, residues of eucalyptol or menthol were not found at all, and only thymol left residues in high concentrations. Residues of thymol found in honey collected from the beehives ranged from 0.75 to 8.20 microg/g for Apilife Var, from 0.03 to 6.30 microg/g for thymol solution in olive oil, and from 0.05 to 6.20 microg/g for thymol solution in ethanol. Even so, natural treatments can be considered to be good alternatives for synthetic acaricides, especially because they do not represent a sanitary risk. PMID- 16366700 TI - Residues of spiroxamine in grapes following field application and their fate from vine to wine. AB - Dissipation of the fungicide spiroxamine in grapes of two vine varieties, Roditis and Cabernet Sauvignon, exposed to field treatments was evaluated. Vines of a grape vineyard located in central Greece were sprayed once or twice with a commercial formulation of the fungicide at 30 g a.i./hL. Residues in grapes, must, and wine were determined by gas chromatography/IT-MS after extraction with cyclohexane-dichloromethane (9:1), with a limit of quantitation 0.02 mg/kg in grapes and 0.012 mg/kg in wine. Under field conditions, spiroxamine dissipation on grapes was faster during the first 2 weeks and then slower to the sixth week. About 7 days after application, half of the initial spiroxamine concentration remained on the grapes; the respective proportion at 42 days was about 10%. At 14 and 35 days, residues were lower than 0.44 and 0.22 mg/kg, respectively, values below the maximum residue levels set by the European Union (1 mg/kg). Spiroxamine residues transferred from grapes into the must and through the vinification process into the wine were also studied. Mean transfer factors of 0.26 and 0.55 were found from grapes into wine for the wines obtained without maceration and with maceration, respectively. Residues in wine, prepared from grapes with a spiroxamine content of 0.11-0.20 mg/kg, varied from <0.026 to 0.09 mg/kg. Spiroxamine diastereomer B was found to dissipate slower than diastereomer A in the field as well as during the vinification process. PMID- 16366701 TI - Fenton's oxidation of food processing wastewater components. Kinetic modeling of protocatechuic acid degradation. AB - The oxidation of protocatechuic acid (PA), a typical phenol-type compound present in food processing wastewater, has been carried out by means of Fenton's reagent. Both the H2O2 and Fe(II) initial concentrations increase the PA degradation rate. Temperature also enhances the PA conversion when raised from 283 to 313 K, a further increase to 323 K results in a lower PA removal. Increasing the PA initial concentration leads to a decrease of conversion values but an opposite effect in terms of removal rate. pH values in the range 3-4 resulted in the total inhibition of the oxidation process. Similar PA depletion rates were experienced regardless of the oxidation state of the catalyst (ferrous or ferric iron). Additionally, an attempt based on the classic Fenton's chemistry plus some other stages accounting for the Fe(II) regeneration from Fe(III) and the inefficient H2O2 decomposition was conducted to model the process. PMID- 16366702 TI - Synthesis of two major toxaphene components and their photostabilities. AB - The synthesis of 2-endo,3-exo,5-endo,6-exo,8,9,10-heptachlorobornane (B7-1001, Hp Sed) and 2-endo,3-exo,5-endo,6-exo,8,9,9,10-octachlorobornane (B8-1412) is described. Both compounds are components of toxaphene, an insecticide that has been widely used in the past. B7-1001 is an important toxaphene congener, comprising up to 99% of total toxaphene concentrations found in fish and sediment samples from treated lakes. B8-1412 is also a significant component of toxaphene contamination in samples from biota. In synthesizing the compounds, 2-exo,3 endo,6-endo,8,9,10-hexachlorobornane (B6-913) was obtained by reduction of the well-known toxaphene component P 32 (B7-515, 2,2,5-endo,6-exo,8,9,10 heptachlorobornane), which was itself isolated from the chlorination products of (+)-camphene. Chlorination of B6-913 provided B7-1001 in 49.5% yield, and P 32 and four other heptachlorobornanes were also detected in the reaction mixture. Structures of two of the heptachlorobornanes were elucidated by MS and NMR as 2 exo,3-endo,6-endo,8,9,9,10-heptachlorobornane (B7-1461) and 2-exo,3,3,6 endo,8,9,10-heptachlorobornane (B7-1303). B8-1412 was isolated from the product mixture obtained by chlorination of 2-exo,3-endo,6-endo,8,9,9,10 heptachlorobornane. Photolysis experiments at lambda = 254 nm revealed that B6 913 is photochemicaly the most stable compound of the seven toxaphene compounds studied, with a t(1/2) of 213 h. B7-1001, having a t(1/2) of 82 h, was the second most stable compound. B8-1412 was degraded more rapidly, with a t(1/2) of 28.8 h, than B7-1001, but was still much more stable than P 50 (B9-1679, 2-endo,3-exo,5 endo,6-exo,8,8,9,10,10-nonachlorobornane), which had a t(1/2) of 9.4 h, despite its reputation as a very persistant compound. Under the same experimental conditions hexachlorobenzene (HLB) and octachlorodibenzodioxine (OCDD) were consumed very quickly with t(1/2) = 0.0025 and 0.0015 h, respectively. PMID- 16366703 TI - Influence of phosphate and copper on reductive dechlorination of thiobencarb in California rice field soils. AB - The potential for reductive dechlorination of the herbicide thiobencarb (TB) by microbes and its prevention in saturated anaerobic rice field soils was examined in laboratory microcosms. TB is effective in controlling both annual grasses and broadleaf weeds. In anoxic microcosms, TB was effectively degraded within 30 days to its dechlorinated product, deschlorothiobencarb (DTB), in two Sacramento Valley rice field soils. TB dechlorination, and subsequent degradation, followed pseudo-zero- (lag phase) and first-order (degradation phase) kinetics. Logistic regression analysis (r2 > 0.841) produced a half-life (t(1/2)) in nonsterile soils ranging from 10 to 15 days, which was also observed when microcosms were amended with low concentrations (<3 mg L(-1)) of copper (Cu2+; as the fungicides Cu(OH)2 and CuSO4.5H2O). High Cu2+ concentrations (>40 mg L(-1)) were added to the microcosms to determine if copper toxicity to dechlorinating microbes is concentration dependent within the range used. After 30 days, the low-copper amended soils closely resembled the nonsterile experiments to which no Cu2+ was added while the high-copper-amended microcosms were similar to the sterile experiment. Microcosms were also separately amended with 5.7 g L(-1) phosphate (PO4(2-); as KH2PO4), a nutrient regularly applied to rice fields. Phosphate amended experiments also showed TB degradation, but no DTB formation, indicating the phosphate played a role, possibly as a microbial inhibitor or an alternative electron acceptor, in limiting the dechlorination of TB. In summary, TB dechlorination was inhibited at high Cu(OH)2, CuSO4.5H2O, and KH2PO4 concentrations. PMID- 16366704 TI - Plant phenolics behave as radical scavengers in the context of insect (Manduca sexta) hemolymph and midgut fluid. AB - To evaluate the prooxidant versus antioxidant properties of plant phenolics toward leaf-feeding caterpillars, quenching of the stable ABTS radical by five phenolics was measured in two physiological contexts: hemolymph and midgut fluid. Addition of tannic acid, chlorogenic acid, quercetin, or catechin to Manduca sexta (L.) gut fluid increased its total antioxidant capacity by 12-45%, with tannic acid and quercetin being the most powerful in this regard. The antioxidant contribution of the phenolics increased with longer (30-60 min) incubation time in gut fluid. Chlorogenic acid and caffeic acid exhibited the weakest antioxidant activity in gut fluid. The total antioxidant capacity of hemolymph is considerably less than that of gut fluid, and in hemolymph chlorogenic and caffeic acids sometimes acted as mild prooxidants, particularly after longer incubation periods (30-60 min), although this trend was not statistically significant. Tannic acid, catechin, and quercetin behaved as antioxidants in hemolymph. These results suggest that many phenolics have radical scavenging activity in the digestive tract, but some may have more detrimental effects after absorption into the hemolymph compartment. PMID- 16366705 TI - Cosorption of atrazine and a lauryl polyoxyethylene oxide nonionic surfactant on smectite. AB - Commercial atrazine formulations commonly contain nonionic surfactants that serve as solubilizing and wetting agents for enhancing the stability and efficacy of the herbicide. The fate of atrazine in soils has been extensively investigated; yet, few studies have considered the effects of formulation components on the fate of atrazine in soils. In this study, we investigated the influence of the nonionic surfactant, Brij 35 (Brij), on the sorption of atrazine on Ca- and K saturated samples of a reference smectite, Panther Creek (PC). In general, Brij concentrations of 50 and 200 mg L(-1) had little effect on atrazine sorption, but sorption was substantially inhibited by Brij concentrations of 2100 mg L(-1). For Brij concentrations of 6300 mg L(-1), atrazine sorption was intermediate between that observed for the 200 and 2100 mg L(-1) Brij systems. Brij molecules themselves were very strongly sorbed by PC, with sorption maxima exceeding 200 g kg(-1). X-ray diffraction analysis of Brij-treated PC indicated that the sorbed Brij was intercalated into interlayers of the smectite. At Brij concentrations of 2100 mg L(-1), Brij competed with atrazine for interlayer sorption sites. In contrast, at the initial Brij concentration of 6300 mg L(-1), the clay interlayers were largely filled with Brij, and excess Brij probably accumulated on external surfaces of the clay as surface micelles. We hypothesize that atrazine partitioning into surfactant micelles on external surfaces of the clay led to enhanced retention by the solid phase. PMID- 16366706 TI - Changes in the concentration of yeast-derived volatile compounds of red wine during malolactic fermentation with four commercial starter cultures of Oenococcus oeni. AB - The effects of malolactic fermentation (MLF) on the concentration of volatile compounds released by yeasts during the production of red wine were investigated by inoculation with four commercial starters of Oenococcus oeni. Volatile compounds in wine at the end of MLF were extracted, analyzed by GC-MS and GC, and compared with those extracted form a noninoculated reference sample. Several esters known to play a role in the aroma profile of red wine, such as C4-C8 ethyl fatty acid esters and 3-methylbutyl acetate, were found to increase with MLF, and their final concentration was dependent on the bacterial starter employed for the induction of MLF. The overall increase of ethyl fatty acid esters was generally larger than the one observed for acetate esters. Ethyl lactate, 3 hydroxybutanoate, 2-phenylethanol, methionol, and gamma-butyrolactone were also increased by bacterial metabolism. The impact of MLF on other volatiles or red wine, including several higher alcohols, fatty acids, and nitrogen compounds, was generally negligible. PMID- 16366707 TI - Characterization of volatile compounds in chilled cod (Gadus morhua) fillets by gas chromatography and detection of quality indicators by an electronic nose. AB - Volatile compounds in cod fillets packed in Styrofoam boxes were analyzed during chilled storage (0.5 degrees C) by gas chromatography (GC)-mass spectrometry and GC-olfactometry to screen potential quality indicators present in concentrations high enough for detection by an electronic nose. Photobacterium phosphoreum dominated the spoilage bacteria on day 12 when the fillets were rejected by sensory analysis. Ketones, mainly 3-hydroxy-2-butanone, were detected in the highest level (33%) at sensory rejection, followed by amines (TMA) (29%), alcohols (15%), acids (4%), aldehydes (3%), and a low level of esters (<1%). The electronic nose's CO sensor showed an increasing response with storage time coinciding with the production of ethanol and 2-methyl-1-propanol that were produced early in the storage, followed by the production of 3-methyl-1-butanol, 3-methyl-butanal, 2,3-butandiol, and ethyl acetate. Lipid-derived aldehydes, like hexanal and decanal, were detected in similar levels throughout the storage time and contributed to the overall sweet odors of cod fillets in combination with other carbonyls (3-hydroxy-2-butanone, acetaldehyde, 2-butanone, 3-pentanone, and 6-methyl-5-heptene-2-one). PMID- 16366708 TI - Chemical composition and antioxidant, antimicrobial, and antifungal activities of the essential oil of Achillea ligustica all. AB - The chemical composition of the essential oil from flowering tops of Achillea ligustica All. was studied. Samples were collected in different localities of Sardinia (Italy) and hydrodistilled both with Clevenger-type and with simultaneous distillation-extraction apparatus. The yields ranged between 0.88 +/ 0.06 and 0.43 +/- 0.02% (vol/dry wt). The essential oils were analyzed by GC-MS, and a total of 96 components were detected. From a qualitative point of view, irrelevant differences between samples were observed. Strong chemical variability depending on the origin of the samples was observed. The major compounds found were santolina alcohol (6.7-21.8%, for the first time detected in A. ligustica), borneol (3.4-20.8%), sabinol (2.1-15.5%), trans-sabinyl acetate (0.9-17.6%), alpha-thujone (0.4-25.8%), and, among sesquiterpenes, viridiflorol (0.7-3.6%). No significant differences were detected between essential oils extracted by hydrodistillation and simultaneous distillation-extraction with CH2Cl2 and n hexane. Antioxidant activity as DPPH radical scavenging activity was expressed in TEAC and ranged between 0.40 and 0.88 mmol/L. The antimicrobial and antifungal activities were investigated on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Penicillium commune, Fusarium oxysporum, Rizoctonia solani, and Aspergillus flavus, showing low activity. PMID- 16366709 TI - Characterization of volatiles in different dry gins. AB - A HS-SPME method coupled to GC-MS was developed and applied for the qualitative and semiquantitative characterization of distilled gin volatile fraction. Sampling, chromatographic conditions, and method performances were evaluated, and the developed method was applied as a comparative study of some of the most popular commercial London Dry Gins and other gins with geographic denominations. During this study, 70 components of the gins' volatile fraction were isolated, tentatively identified or identified by reference compounds. They were mainly represented by mono- and sesquiterpenic compounds, which were quantitatively determined. The comparative study of London Dry Gins and gins with geographical indication permitted clear differentiation between the gins with geographical indication and one of the London Dry Gins tested. PMID- 16366710 TI - Haze formation in model beer systems. AB - The interaction of a haze-active protein (gliadin) and a haze-active polyphenol (tannic acid) was studied in a model beer system in order to investigate the principle mechanisms of haze formation at low temperatures. Low concentrations (g/L) of tannic acid, high concentrations of gliadin, and comparatively high temperatures lead to maximum haze values. When considered on a molar basis, the greatest haze levels are displayed at an approximate 1:1 equivalence of polyphenol and protein. The greater part of haze formation was completed within 0.5 h, irrespective of the concentration of gliadin, the concentration of tannic acid, and the temperature of the model solution. PMID- 16366711 TI - Tailoring of Atlantic salmon (Salmo salar L.) flesh lipid composition and sensory quality by replacing fish oil with a vegetable oil blend. AB - Atlantic salmon (Salmo salar L.) juveniles were fed either 100% fish oil (FO), 75% vegetable oil (VO), or 100% VO throughout their life cycle to harvest weight followed by a finishing diet period when all groups were fed 100% FO. The two experimental VO diets were tested at two different locations (Scotland and Norway) against the same control diet (100% FO). The VO blend was composed of rapeseed oil, palm oil, and linseed oil using capelin oil as a control for fatty acid class compositions. Flesh fatty acid profiles were measured regularly throughout the experiment, with the times of sampling determined by changes in pellet size/lipid content and fish life stage. Growth and mortality rates were not significantly affected by dietary fatty acid compositions throughout the life cycle, except during the seawater winter period in Norway when both growth and protein utilization were increased in salmon fed 100% VO compared to 100% FO. Flesh fatty acid composition was highly influenced by that of the diet, and after the finishing diet period the weekly intake recommendations of very long chain n 3 polyunsaturated fatty acid (VLCn-3 PUFA) for human health were 80 and 56% satisfied by a 200 g meal of 75% VO and 100% VO flesh, respectively. No effect on flesh astaxanthin levels was observed in relation to changing dietary oil sources. Sensory evaluation showed only minor differences between salmon flesh from the dietary groups, although prior to the finishing diet period, flesh from 100% VO had less rancid and marine characteristics and was preferred over flesh from the other dietary groups by a trained taste panel. After the finishing diet period, the levels of typical vegetable oil fatty acids in flesh were reduced, whereas those of VLCn-3 PUFA increased to levels comparable with a 100% FO fed salmon. No differences in any of the sensory characteristics were observed between dietary groups. By blending VOs to provide balanced levels of dietary fatty acids, up to 100% of the fish oil can be replaced by the VO blend without compromising growth or flesh quality. At the same time, 75% of the dietary fish oil can be replaced without compromising flesh VLCn-3 PUFA content, thereby providing a beneficial nutritional profile for human consumption. PMID- 16366712 TI - Binding of the pepper alkaloid piperine to bovine beta-lactoglobulin: circular dichroism spectroscopy and molecular modeling study. AB - The pepper alkaloid piperine is a nontoxic, natural dietary compound with a broad range of physiological activity. The present work is the first demonstration of its interaction with a mammalian protein. Circular dichroism (CD) spectroscopy was used to reveal and analyze the binding of piperine to a lipocalin protein. Induced CD spectra measured in pH 7.7 phosphate buffer at 37 degrees C demonstrated reversible, non-covalent association of piperine with bovine beta lactoglobulin (BLG), the major whey protein in milk. The binding parameters (K(a) approximately 8 x 10(4) M(-1), n = 0.8) determined from the CD titration data showed no significant differences between the piperine binding properties of the two main genetic variants of BLG (A and B). The vanishing extrinsic CD signal obtained upon acidification of the piperine-BLG sample solution (Tanford transition) suggested that the ligand binds in the central hydrophobic cavity of the beta-barrel. The cavity binding concept was further supported by a CD displacement experiment using palmitic acid, the well-known hydrophobic ligand of BLG. Molecular docking calculations showed that piperine can be efficiently accommodated within the calyx of BLG. Additional molecular modeling calculations indicated that the beta-barrel of human tear lipocalin, human serum retinol binding protein, and human neutrophil gelatinase associated lipocalin might also accommodate a piperine molecule. PMID- 16366713 TI - Elicitation of resveratrol in peanut kernels by application of abiotic stresses. AB - Application of no postharvest stress or by size reduction, grinding, chopping, or slicing and exposure to UV light or ultrasound were investigated for the elicitation of trans-resveratrol, total phenolic compounds, and antioxidant activity (AOA) in peanut kernels. AOA and total phenolic compounds did not increase in stressed peanuts over incubation times 0-48 h at 25 degrees C; however, an increase in trans-resveratrol concentration over time occurred. Slicing (2 mm), ultrasound exposure for 4 min at 25 degrees C, and incubation for 36 h produced the highest level of trans-resveratrol synthesis in peanuts, increasing concentrations from 0.48 +/- 0.08 microg/g in untreated peanuts to 3.96 +/- 0.96 microg/g. AOA in stressed peanuts was negatively correlated with trans-resveratrol concentration, indicating that as AOA decreased trans resveratrol concentration increased. PMID- 16366714 TI - Morphological, structural, thermal, and rheological characteristics of starches separated from apples of different cultivars. AB - The starches were separated from unripe apples of five cultivars (Criterion, Ruspippum, Red Spur, Skyline Supreme, and Granny Smith) and evaluated using scanning electron microscopy (SEM), gel permeation chromatography (GPC), X-ray diffraction, differential scanning calorimetry (DSC), and dynamic viscoelasticity. SEM showed the presence of round granules as well as granules that had been partially degraded, probably by amylases. The starch granules in different apple starches ranged between 4.1 and 12.0 mum. Debranching of starch with isoamylase and subsequent fractionation of debranched materials by GPC revealed the presence of an apparent amylose, an intermediate fraction (mixture of amylose and amylopectin), long side chains of amylopectin, and short side chains of amylopectin in the range of 28-35.2, 3.6-4.4, 20-21.3, and 39.9-47.1%, respectively. The swelling power of starches ranged between 14.4 and 21.3 g/g. X ray diffraction of apple starches showed a mixture of A- and B-type patterns. All apple starches showed peak intensities lower than that observed for normal corn and potato starch, indicating the lower crystallinity. The transition temperatures (onset temperature, T(o); peak temperature, T(p); and conclusion temperature, T(c)) and enthalpy of gelatinization (deltaH(gel)) determined using DSC ranged between 54.7 and 56.2 degrees C, between 57.1 and 59.1 degrees C, between 60.2 and 63.5 degrees C, and between 3.3 and 4.2 J/g, respectively. The viscoelastic properties of starch from different cultivars measured during heating and cooling using a rheometer differed significantly. Red Spur and Criterion starches with larger granule size showed higher G' and G' ' values, whereas those containing smaller size and amylolytically degraded granules showed lower G' and G' '. PMID- 16366715 TI - Determination of the secondary structure of Kluyveromyces lactis beta galactosidase by circular dichroism and its structure-activity relationship as a function of the pH. AB - The secondary structure of Kluyveromyces lactis beta-galactosidase was determined by circular dichroism. It is mainly a beta-type protein, having 22% beta-turns, 14% parallel beta-sheet, 25% antiparallel beta-sheet, 34% unordered structure, and only 5% alpha-helix. The structure-activity relationship as a function of the pH was also studied. The pH conditions leading to the highest secondary structure content (100% ellipticity) of the enzyme was found at pH 7.0; at pH 6.5-7.0, the percent ellipticity decreased slightly, suggesting little structural change, but the activity decreased significantly, probably because of variations in critical residues. On the other hand, at pH's above 7.0, a more noticeable change in ellipticity was observed due to structural changes; the CD analysis showed a small increase in the helical content toward higher pH, whereas the maximum activity was found at pH 7.5, meaning that the changes produced in the secondary structure at this pH favored the interaction between the enzyme and the substrate. PMID- 16366716 TI - Evaluation of the site specific protein glycation and antioxidant capacity of rare sugar-protein/peptide conjugates. AB - Protein-sugar conjugates generated in nonenzymatic glycation of alpha-lactalbumin (LA) with rare sugars [D-allose (All) and D-psicose (Psi)] and alimentary sugars as controls [D-glucose (Glc) and D-fructose (Fru)] were qualitatively determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Mass spectra revealed that the extent of glycation at lysine residues on LA with D-aldose molecules was very much higher than that of glycation with d-ketose molecules. To identify the specific site of glycation, the peptide mapping was established from protease V8 digestion, using a combination of computational cutting of proteins and MALDI-TOF-MS. As compared to peptide mapping, three and seven glycation sites were located in the primary structure of LA-ketose and LA-aldose conjugates, respectively. On the other hand, the antioxidant activities of protein-sugar conjugates and their peptic hydrolysates were investigated by 1,1-diphenyl-2-picrylhydrazyl radical scavenging method. The antioxidant activities of proteins/peptides glycated with rare sugars were significantly higher than those modified with the control sugars. The results indicated that the glycation degree and position were not markedly different between rare sugar and corresponding control sugar, but the antioxidant properties of protein and its hydrolysate were significantly enhanced by modifying with rare sugar. PMID- 16366717 TI - The effect of thermal history on the maillard reaction in a glassy matrix. AB - To test whether the extent of physical aging affected the reaction rate, Maillard reaction kinetics were studied in glassy model preservation systems subjected to two different thermal histories. The glass transition temperature and physical aging of the matrix were determined using differential scanning calorimetry, and the normalized heat capacities were modeled using the Tool-Narayanaswamy-Moynihan approach. Samples prepared using the different thermal histories initially had different degrees of aging, but these were practically indistinguishable after 10 h under the reaction conditions (65 degrees C); the samples underwent rapid structural relaxation at that temperature. The reaction of glucose and lysine in an amorphous trehalose/sucrose matrix was followed using spectrophotometric and chromatographic analysis. A difference in reaction rate could only be distinguished in the rate of consumption of glucose, which was approximately 20% faster in the minimally aged matrix; no significant differences were seen in any other indicator of reaction. PMID- 16366718 TI - Immobilized zinc affinity chromatography of pectin hydroxamic acids for purification of trypsin inhibitors from soybean and sweet potato. AB - Commercial pectin (with a 94% degree of esterification, DE94) suspended in methanol was reacted with methanolic alkaline hydroxylamine at room temperature for 20 h to prepare pectin hydroxamic acids (PHAs). The prepared PHA was coupled to the epoxy-activated Sepharose 6B gel to get immobilized PHA resins. The immobilized PHA resin was then balanced in column with 2 mM ZnCl2 in 50 mM Tris HCl buffer (pH 7.9) to test the immobilized Zn-PHA gel as solid phase for immobilized metal affinity chromatography for the purification of trypsin inhibitors (TIs) from soybean and sweet potato. Using TI activity staining, it was found that purified TIs from the commercial soybean and sweet potato after trypsin affinity column purification could be adsorbed onto an immobilized Zn-PHA affinity column and eluted by 100 mM EDTA in 10 mM Tris-HCl buffer (pH 7.9). The immobilized Zn-PHA affinity column was used for TI purifications from crude extracts of sweet potato. The recovery of TI activity for one step was 90%, with 19.74-fold purification increase. PMID- 16366719 TI - Purification and characterization of Ocimum basilicum L. polyphenol oxidase. AB - A partial characterization of polyphenol oxidase (PPO) activity in Ocimum basilicum L. is described. PPO in O. basilicum L. was extracted and purified through (NH4)2SO4 precipitation, dialysis, and a Sepharose 4B-l-tyrosine-p aminobenzoic acid affinity column. The samples obtained from (NH4)2SO4 precipitation and dialysis were used for the characterization of PPO. At the end of purification by affinity chromatography, 11.5-fold purification was achived. The purified enzyme exhibited a clear single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular mass of the enzyme was estimated to be approximately 54 kDa. The contents of total phenolic and protein of O. basilicum L. extracts were determined. The total phenolic content of O. basilicum L. was determined spectrophotometrically according to the Folin Ciocalteu procedure and was found to be 280 mg 100 g(-1) on a fresh weight basis. The protein content was determined according to the Bradford method. The enzyme showed activity to 4-methylcatechol, catechol, and pyrogallol substrates, but not to tyrosine. Therefore, of these three substrates, 4-methylcatecol was the best substrate due to the highest V(max)/K(m) value, followed by pyrogallol and catechol. The optimum pH was at 6, 8, and 9 for 4-methylcatechol, catechol, and pyrogallol, respectively. The enzyme had an optimum temperature of 20, 40, and 50 degrees C for 4-methylcatechol, catechol, and pyrogallol, respectively. It was found that optimum temperature and pH were dependent on the substrates studied. The enzyme activity with increasing temperature and inactivation time for 4 methylcatechol, catechol, and pyrogallol substrates decreased due to heat denaturation of the enzyme. PMID- 16366720 TI - Pro-oxidative characteristics of trout hemoglobin and myoglobin: a role for released heme in oxidation of lipids. AB - The molecular mass of trout myoglobin was 16017 Da based on electrospray ionization mass spectrometry. A Root effect (low oxygen affinity at pH 6.3) was determined in trout hemoglobin but not myoglobin. At pH 6.3, myoglobin autoxidized more rapidly (3.5-fold) as compared to anodic hemoglobin. Anodic hemoglobin was a better catalyst of lipid oxidation in washed cod muscle as compared to myoglobin at pH 6.3. This suggested that some process other than met heme protein formation was the rate-limiting step in lipid oxidation processes. Heme loss rates were determined using the apomyoglobin mutant H64Y prepared from sperm whale. Anodic hemoglobin released its heme group much more rapidly than myoglobin. In comparisons of anodic and cathodic hemoglobins, heme loss rate better predicted the onset of lipid oxidation than autoxidation rate. These studies collectively suggest that heme dissociation has a primary role in the ability of different heme proteins to promote lipid oxidation processes. PMID- 16366721 TI - Degradation of endogenous and exogenous genes of roundup-ready soybean during food processing. AB - Roundup-Ready soybeans have been genetically modified to resist the effects of the herbicidal glyphosate and have become the most prevalent transgenic crop in the world. In this work, Roundup-Ready soybeans were used as raw material to study the effects of critical processing procedures such as grinding, cooking, blending, homogenization, sterilization, and spray-drying on the length of DNA fragments of an endogenous gene (lectin) and an exogenous gene (epsps) examined in material from three soybean foods of bean curd, soy milk, and soy powder and from samples taken during their processing. The results showed that various processing procedures caused degradations of both the endogenous and exogenous genes to different degrees. In the grinding procedure, endogenous gene DNA was degraded from 1883 to approximately 836 bp, and exogenous gene DNA was degraded from 1512 to approximately 408 bp. In the blending and squeeze-molding procedures, exogenous gene DNA was also degraded from about 408 to 190 bp, but there was no obvious action on the endogenous gene. After the endogenous and exogenous genes had been degraded to some degree, such as 836 and 408 bp, respectively, they were not evidently affected by cooking procedure at 100 degrees C for 15 min. However, the endogenous gene was further considerably degraded from around 836 to 162 bp in the sterilization procedure at 121 degrees C for 30 s. The effect of the homogenization step on endogenous and exogenous genes was similar to that of the cooking procedure. The coagulation procedure, principally a biochemical reaction, did not greatly affect the exogenous gene but did affect endogenous gene, reducing DNA size from about 836 to 407 bp. Furthermore, the spray-drying procedure, a process of physical shearing, high temperature, and sudden high pressure, distinctly caused degradation of both the lectin and epsps genes, rapidly decreasing the sizes from about 836 to 162 bp for the endogenous gene and from about 408 to 190 bp for the exogenous gene. PMID- 16366722 TI - Alkylating potential of potassium sorbate. AB - A kinetic study of the alkylating potential of potassium sorbate (S)-a food preservative used worldwide-in 7:3 water/dioxane medium was performed. The following conclusions were drawn: (i) Potassium sorbate shows alkylating activity on the nucleophile 4-(p-nitrobenzyl)pyridine (NBP), a trap for alkylating agents with nucleophilic characteristics similar to those of DNA bases, (ii) The NBP alkylation reaction complies with the rate equation r = k(alk)[H+][S][NBP]/(K(a) + [H+]), K(a) being the sorbic acid dissociation constant and k(alk) the rate constant of NBP alkylation by the undissociated acid. In the range of pH 5-6, the alkylation time ranges between 18 days (pH 5.2) and >1 month (pH > or = 6). (iii) NBP alkylation occurs through a reaction with deltaH# = 78 kJ mol(-1), which is much higher than those of NBP alkylation by stronger alkylating agents. (iv) The absorption coefficient of the sorbate-NBP adduct was determined to be epsilon = 204 M(-1) cm(-1) (lambda = 580 nm), this value being rationalized in terms of the adduct structure. (v) The results can help to establish suitable expiration times for products preserved with potassium sorbate. PMID- 16366723 TI - Antioxidant activity of cysteine, tryptophan, and methionine residues in continuous phase beta-lactoglobulin in oil-in-water emulsions. AB - Proteins dispersed in the continuous phase of oil-in-water emulsions are capable of inhibiting lipid oxidation reactions. The antioxidant activity of these proteins is thought to encompass both free radical scavenging by amino acid residues and chelation of prooxidative transition metals; however, the precise mechanism by which this occurs remains unclear. In this study, the oxidative stability of cysteine, tryptophan, and methionine residues in continuous phase beta-lactoglobulin (beta-Lg) in a Brij-stabilized menhaden oil-in-water emulsion was determined. The presence of low concentrations of continuous phase beta-Lg (250 and 750 microg/mL) significantly inhibited lipid oxidation as determined by lipid hydroperoxides and thiobarbituric acid reactive substances analysis. It was observed that cysteine oxidized before tryptophan in beta-Lg, and both residues oxidized before lipid oxidation could be detected. No oxidation of the methionine residues of beta-Lg was observed despite its reported high oxidative susceptibility. It is conceivable that surface exposure of amino acid residues greatly affects their oxidation kinetics, which may explain why some residues are preferentially oxidized relative to others. Further elucidation of the mechanisms governing free radical scavenging of amino acids could lead to more effective applications of proteins as antioxidants within oil-in-water food emulsions. PMID- 16366724 TI - Strecker type degradation of phenylalanine by 4-hydroxy-2-nonenal in model systems. AB - The reaction of 4-hydroxy-2-nonenal, an oxidative stress product, with phenylalanine in acetonitrile-water (2:1, 1:1, and 1:2) at 37, 60, and 80 degrees C was investigated to determine whether 4-hydroxy-2-alkenals degrade amino acids, analogously to 4,5-epoxy-2-alkenals, and to compare the reactivities of both hydroxyalkenals and epoxyalkenals for production of Strecker aldehydes. In addition to the formation of N-substituted 2-pentylpyrrole and 2-pentylfuran, the studied hydroxyalkenal also degraded phenylalanine to phenylacetaldehyde with a reaction yield of 17%. The reaction mechanism is suggested to be produced through the corresponding imine, which is then decarboxylated and hydrolyzed. This reaction also produced a conjugated amine, which both may be one of the origins of the produced 2-pentyl-1H-pyrrole and may contribute to the development of browning in these reactions. 4-Hydroxy-2-nonenal and 4,5-epoxy-2-decenal degraded phenylalanine in an analogous extent, which is likely a consequence of the similarity of the degradation mechanisms involved. These results suggest that different lipid oxidation products are able to degrade amino acids; therefore, the Strecker type degradation of amino acids produced by oxidized lipids may be quantitatively significant in foods. PMID- 16366725 TI - Antioxidant and pro-oxidant activities of aqueous extracts and crude polyphenolic fractions of rooibos (Aspalathus linearis). AB - Unfermented rooibos tea is known to contain higher levels of total polyphenols and flavonoids than its fermented counterpart, making it the obvious choice for the preparation of flavonoid-enriched fractions. Evaluation of aqueous extracts and crude polyphenolic fractions of unfermented and fermented rooibos showed anti and/or pro-oxidant activities, using a linoleic acid-Tween-buffer emulsion for lipid peroxidation and the deoxyribose degradation assay, based on a Fenton reaction model system containing FeCl3-EDTA and H2O2 for the generation of hydroxyl radicals. Except for the ethyl acetate fraction, with the highest total polyphenol (TP) content and offering the least protection presumably due to pro oxidant activity, the inhibition of lipid peroxidation by the samples correlated moderately with their TP content in a linear relationship (r = 0.896, P < 0.01). Using the deoxyribose degradation assay, the pro-oxidant activity of the aqueous extracts and their crude polymeric fractions (0.1 mg/mL in the reaction mixture) was linear with respect to their dihydrochalcone (aspalathin and nothofagin) (r = 0.977, P = 0.023) and flavonoid (r = 0.971, P = 0.029) content. Pro-oxidant activity was demonstrated for pure aspalathin. Using the same assay, but with ascorbate added to regenerate Fe3+ to Fe2+, the aqueous extract and crude polymeric fraction of fermented rooibos displayed hydroxyl radical scavenging activity. Fermentation (i.e., oxidation) of rooibos decreased the pro-oxidant activity of aqueous extracts, which was contributed to a decrease in their dihydrochalcone content. The in vitro pro-oxidant activity displayed by flavonoid enriched fractions of rooibos demonstrates that one must be aware of the potential adverse biological properties of potent antioxidant extracts utilized as dietary supplements. PMID- 16366726 TI - Detection of transgenic and endogenous plant DNA fragments in the blood, tissues, and digesta of broilers. AB - The aim was to determine the fate of transgenic and endogenous plant DNA fragments in the blood, tissues, and digesta of broilers. Male broiler chicks (n = 24) were allocated at 1 day old to each of four treatment diets designated T1 T4. T1 and T2 contained the near isogenic nongenetically modified (GM) maize grain, whereas T3 and T4 contained GM maize grain [cry1a(b) gene]; T1 and T3 also contained the near isogenic non-GM soybean meal, whereas T2 and T4 contained GM soybean meal (cp4epsps gene). Four days prior to slaughter at 39-42 days old, 50% of the broilers on T2-T4 had the source(s) of GM ingredients replaced by their non-GM counterparts. Detection of specific DNA sequences in feed, tissue, and digesta samples was completed by polymerase chain reaction analysis. Seven primer pairs were used to amplify fragments ( approximately 200 bp) from single copy genes (maize high mobility protein, soya lectin, and transgenes in the GM feeds) and multicopy genes (poultry mitochondrial cytochrome b, maize, and soya rubisco). There was no effect of treatment on the measured growth performance parameters. Except for a single detection of lectin (nontransgenic single copy gene; unsubstantiated) in the extracted DNA from one bursa tissue sample, there was no positive detection of any endogenous or transgenic single copy genes in either blood or tissue DNA samples. However, the multicopy rubisco gene was detected in a proportion of samples from all tissue types (23% of total across all tissues studied) and in low numbers in blood. Feed-derived DNA was found to survive complete degradation up to the large intestine. Transgenic DNA was detected in gizzard digesta but not in intestinal digesta 96 h after the last feeding of treatment diets containing a source of GM maize and/or soybean meal. PMID- 16366727 TI - Meat and ascorbic acid can promote Fe availability from Fe-phytate but not from Fe-tannic acid complexes. AB - This study utilized an in vitro digestion/Caco-2 cell model to determine the levels of ascorbic acid (AA) and "meat factor" needed to promote Fe absorption from Fe complexed with phytic acid (PA) or tannic acid (TA). AA reversed the inhibition of Fe absorption by PA beginning at a molar ratio of 1:20:1 (Fe:PA:AA) but essentially had no effect on the Fe complexed with TA. Fish also reversed the inhibition of Fe uptake by PA but not by TA. TA and fish decreased total Fe solubility. Iron in the presence of PA was highly soluble. AA, but not fish, increased the percentage of soluble Fe as Fe2+ in the presence of both inhibitors. The results indicate that monoferric phytate is a form of Fe that can be available for absorption in the presence of uptake promoters. In contrast, a TA-Fe complex is much less soluble and unavailable in the presence of promoters. PMID- 16366728 TI - Phytate, calcium, iron, and zinc contents and their molar ratios in foods commonly consumed in China. AB - A total of 60 food samples commonly consumed in China were analyzed for phytate using the anion-exchange method and for calcium, iron, and zinc using atomic absorption spectrophotometry. The foods analyzed included those based on cereal grains and soybean. Phytate contents expressed on a wet weight basis ranged from 0 for foods made from starches to 1878 mg/100 g for dried stick-shaped soybean milk film. The calcium contents were between 2.08 mg/100 g for ground corn and 760.67 mg/100 g for diced fried soybean curd. The lowest values of iron and zinc were 0.04 mg/100 g for Panjin pearl rice cooked with discarding extra water and 0.08 mg/100 g for potato and bean starches, while the highest values of iron and zinc were observed in dried stick-shaped soybean milk film. Although many foods were relatively rich in calcium, zinc, and iron, many also contained a higher level of phytate. Of the 60 food samples, 34 foods had a phytate/calcium molar ratio >0.24, 53 foods had a phytate/iron molar ratio >1, 31 foods had a phytate/zinc molar ratio >15, and only 7 foods had a phytate x calcium/zinc >200. Phytate in foods impair the bioavailability of calcium, iron, and zinc, which to some extent depends upon food processing and cooking methods. PMID- 16366729 TI - Wild artichoke prevents the age-associated loss of vasomotor function. AB - Endothelial dysfunction, which is more often observed in conduit arteries such as the aorta, carotid, femoral, and brachial arteries, is largely due to alterations in cellular signal transduction initiated by an escalating cycle of damage triggered by oxidative stress. This phenomenon is exacerbated in the elderly, where a progressive loss of vascular endothelial function and concurrent loss of vasomotor control is frequent. In a previous study, we demonstrated that the wild artichoke (Cynara cardunculus) is able to increase the production of the vasorelaxant factor nitric oxide by cultured aortic endothelial cells. We now extended that study to verify (1) the vasorelaxant potential of C. cardunculus on isolated rat aortic rings and (2) whether the vasomodulating properties of C. cardunculus are maintained in vivo, after administration to aged rats. The results demonstrate that the wild artichoke and its main components, namely, luteolin and apigenin, improve aortic relaxation when added to the incubation bath. Moreover, the feeding of wild artichoke [10 mg (kg of polyphenols)(-1) day( 1)] to aged rats significantly restores proper vasomotion, to a degree similar to that observed in young animals. This study provides further justification to the advice to consume wild greens as part of a balanced diet and suggests that close attention should be paid to the diet of the elderly, because it can effectively modulate important parameters of cardiovascular risk. PMID- 16366730 TI - Transformation of organoarsenical species by the microflora of freshwater crayfish. AB - A study of the transformation of arsenic species by the microflora of the freshwater crayfish Procambarus clarkii was carried out. The study of the degradation of AB (arsenobetaine) was performed in aerobic conditions in two culture media (tryptic soy broth and saline medium) at two temperatures (30 and 8 degrees C). The microflora transformed AB into TMAO (trimethylarsine oxide), DMA (dimethylarsinate), MA (methylarsonate), and an unidentified compound (U1). The quickest transformations were carried out by microflora from hepatopancreas incubated in saline medium at 30 degrees C. The individualized study of other arsenic species [AC (arsenocholine), TETRA (tetramethylarsonium ion), TMAO, DMA, and MA] was also performed in saline medium. The only transformation observed was of AC into AB. The bacteria possibly responsible for AB degradation were isolated, identified by phenotypic and genotypic methods, and individually assayed for AB transformation. Only isolates allocated to the species Pseudomonas putida were able to metabolize AB. PMID- 16366731 TI - Suppressive effects of ethanolic extracts from propolis and its main botanical origin on dioxin toxicity. AB - Suppressive effects of ethanolic extracts prepared from propolis group 12 and its main botanical origin (leaf bud of Baccharis dracunculifolia) on transformation of the aryl hydrocarbon receptor (AhR), the initial action of dioxin toxicity, were investigated. It was found that suppressive effects of propolis on AhR transformation were relatively higher than those of resins of its botanical origin in cell-free system and in Hepa-1c1c7 cells. When the composition of chemical ingredients was measured, propolis contained slightly higher amounts of flavonoid aglycones as compared with its botanical origin with the same characteristics. Moreover, antiradical activity, one of the typical biological activities of flavonoids, in propolis was also slightly higher than that in its botanical origin. These results indicate that not only propolis but also its botanical origin contains high amounts of flavonoid aglycones and that both of them are useful dietary sources for flavonoids with a potency to prevent dioxin toxicity. PMID- 16366734 TI - Activation of NK cells by an endocytosed receptor for soluble HLA-G. AB - Signaling from endosomes is emerging as a mechanism by which selected receptors provide sustained signals distinct from those generated at the plasma membrane. The activity of natural killer (NK) cells, which are important effectors of innate immunity and regulators of adaptive immunity, is controlled primarily by receptors that are at the cell surface. Here we show that cytokine secretion by resting human NK cells is induced by soluble, but not solid-phase, antibodies to the killer cell immunoglobulin-like receptor (KIR) 2DL4, a receptor for human leukocyte antigen (HLA)-G. KIR2DL4 was constitutively internalized into Rab5 positive compartments via a dynamin-dependent process. Soluble HLA-G was endocytosed into KIR2DL4-containing compartments in NK cells and in 293T cells transfected with KIR2DL4. Chemokine secretion induced by KIR2DL4 transfection into 293T cells occurred only with recombinant forms of KIR2DL4 that trafficked to endosomes. The profile of genes up-regulated by KIR2DL4 engagement on resting NK cells revealed a proinflammatory/proangiogenic response. Soluble HLA-G induced secretion of a similar set of cytokines and chemokines. This unique stimulation of resting NK cells by soluble HLA-G, which is endocytosed by KIR2DL4, implies that NK cells may provide useful functions at sites of HLA-G expression, such as promotion of vascularization in maternal decidua during early pregnancy. PMID- 16366735 TI - Dynamic remodeling of dendritic arbors in GABAergic interneurons of adult visual cortex. AB - Despite decades of evidence for functional plasticity in the adult brain, the role of structural plasticity in its manifestation remains unclear. To examine the extent of neuronal remodeling that occurs in the brain on a day-to-day basis, we used a multiphoton-based microscopy system for chronic in vivo imaging and reconstruction of entire neurons in the superficial layers of the rodent cerebral cortex. Here we show the first unambiguous evidence (to our knowledge) of dendrite growth and remodeling in adult neurons. Over a period of months, neurons could be seen extending and retracting existing branches, and in rare cases adding new branch tips. Neurons exhibiting dynamic arbor rearrangements were GABA positive non-pyramidal interneurons, while pyramidal cells remained stable. These results are consistent with the idea that dendritic structural remodeling is a substrate for adult plasticity and they suggest that circuit rearrangement in the adult cortex is restricted by cell type-specific rules. PMID- 16366737 TI - Neurodegeneration from mitochondrial insufficiency: nutrients, stem cells, growth factors, and prospects for brain rebuilding using integrative management. AB - Degenerative brain disorders (neurodegeneration) can be frustrating for both conventional and alternative practitioners. A more comprehensive, integrative approach is urgently needed. One emerging focus for intervention is brain energetics. Specifically, mitochondrial insufficiency contributes to the etiopathology of many such disorders. Electron leakages inherent to mitochondrial energetics generate reactive oxygen free radical species that may place the ultimate limit on lifespan. Exogenous toxins, such as mercury and other environmental contaminants, exacerbate mitochondrial electron leakage, hastening their demise and that of their host cells. Studies of the brain in Alzheimer's and other dementias, Down syndrome, stroke, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, Huntington's disease, Friedreich's ataxia, aging, and constitutive disorders demonstrate impairments of the mitochondrial citric acid cycle and oxidative phosphorylation (OXPHOS) enzymes. Imaging or metabolic assays frequently reveal energetic insufficiency and depleted energy reserve in brain tissue in situ. Orthomolecular nutrients involved in mitochondrial metabolism provide clinical benefit. Among these are the essential minerals and the B vitamin group; vitamins E and K; and the antioxidant and energetic cofactors alpha-lipoic acid (ALA), ubiquinone (coenzyme Q10; CoQ10), and nicotinamide adenine dinucleotide, reduced (NADH). Recent advances in the area of stem cells and growth factors encourage optimism regarding brain regeneration. The trophic nutrients acetyl L-carnitine (ALCAR), glycerophosphocholine (GPC), and phosphatidylserine (PS) provide mitochondrial support and conserve growth factor receptors; all three improved cognition in double-blind trials. The omega-3 fatty acid docosahexaenoic acid (DHA) is enzymatically combined with GPC and PS to form membrane phospholipids for nerve cell expansion. Practical recommendations are presented for integrating these safe and well-tolerated orthomolecular nutrients into a comprehensive dietary supplementation program for brain vitality and productive lifespan. PMID- 16366736 TI - Sirt1 regulates insulin secretion by repressing UCP2 in pancreatic beta cells. AB - Sir2 and insulin/IGF-1 are the major pathways that impinge upon aging in lower organisms. In Caenorhabditis elegans a possible genetic link between Sir2 and the insulin/IGF-1 pathway has been reported. Here we investigate such a link in mammals. We show that Sirt1 positively regulates insulin secretion in pancreatic beta cells. Sirt1 represses the uncoupling protein (UCP) gene UCP2 by binding directly to the UCP2 promoter. In beta cell lines in which Sirt1 is reduced by SiRNA, UCP2 levels are elevated and insulin secretion is blunted. The up regulation of UCP2 is associated with a failure of cells to increase ATP levels after glucose stimulation. Knockdown of UCP2 restores the ability to secrete insulin in cells with reduced Sirt1, showing that UCP2 causes the defect in glucose-stimulated insulin secretion. Food deprivation induces UCP2 in mouse pancreas, which may occur via a reduction in NAD (a derivative of niacin) levels in the pancreas and down-regulation of Sirt1. Sirt1 knockout mice display constitutively high UCP2 expression. Our findings show that Sirt1 regulates UCP2 in beta cells to affect insulin secretion. PMID- 16366738 TI - Adverse effects of dietary fructose. AB - The consumption of fructose, primarily from high-fructose corn syrup (HFCS), has increased considerably in the United States during the past several decades. Intake of HFCS may now exceed that of the other major caloric sweetener, sucrose. Some nutritionists believe fructose is a safer form of sugar than sucrose, particularly for people with diabetes mellitus, because it does not adversely affect blood-glucose regulation, at least in the short-term. However, fructose has potentially harmful effects on other aspects of metabolism. In particular, fructose is a potent reducing sugar that promotes the formation of toxic advanced glycation end-products, which appear to play a role in the aging process; in the pathogenesis of the vascular, renal, and ocular complications of diabetes; and in the development of atherosclerosis. Fructose has also been implicated as the main cause of symptoms in some patients with chronic diarrhea or other functional bowel disturbances. In addition, excessive fructose consumption may be responsible in part for the increasing prevalence of obesity, diabetes mellitus, and non-alcoholic fatty liver disease. Although the long-term effects of fructose consumption have not been adequately studied in humans, the available evidence suggests it may be more harmful than is generally recognized. The extent to which a person might be adversely affected by dietary fructose depends both on the amount consumed and on individual tolerance. With a few exceptions, the relatively small amounts of fructose that occur naturally in fruits and vegetables are unlikely to have deleterious effects, and this review is not meant to discourage the consumption of these healthful foods. PMID- 16366739 TI - Seasonal variations of selected cardiovascular risk factors. AB - This article reviews research on selected biomarkers of cardiovascular risk - cholesterol and other lipids, C-reactive protein (CRP), fibrinogen, homocysteine in the attempt to determine the existence of a predictable seasonal chronobiological pattern of variation. Studies dating as far back as the 1930s have reported seasonal variations in cholesterol levels. Statistically significant seasonal changes in lipid levels have been found in individuals irrespective of the country where the research has been conducted, and irrespective of the age, sex, ethnicity, and baseline lipid levels of the study subjects. While not all studies have been in complete agreement on either the amplitude (degree of seasonal change) or month/s of highest lipid levels, a strong winter/summer difference has been found in most studies. Existing evidence for an independent effect of season in variation of CRP is weak. Studies have consistently reported significant seasonal variations in fibrinogen levels. While other biological factors clearly interact to affect fibrinogen variability, seasonality appears to be an independent source of variability. Evidence from several studies points to a lack of seasonal variability in homocysteine levels. Although seasonal variability is just one source of periodicity influencing biological function and assessments in clinical practice, for some biomarkers, including lipids and fibrinogen, it is a source of variability that warrants consideration prior to a decision to treat and in assessing response to interventions. PMID- 16366740 TI - The effect of an herbal supplement containing black tea and caffeine on metabolic parameters in humans. AB - OBJECTIVE: The objective of this study was to test an herbal supplement containing black tea (the fully oxidized form of Camellia sinensis) and caffeine for stimulation of thermogenesis. METHODS/MATERIALS: A double-blind, placebo controlled, crossover study was conducted on 16 healthy, weight-stable, non smoking subjects, ages 21-55 years, with body mass index (BMI) of 20-30 kg/m2, and on no medications other than oral contraceptives or hormone replacement therapy. Subjects had no caffeine for 48 hours, no exercise for 24 hours, and no food for 12 hours before each visit. Area under the curve (AUC) for resting metabolic rate (RMR), respiratory quotient (RQ), blood pressure, pulse rate, and temperature were measured. At each visit RMR was measured at baseline and at one and two hours following oral administration of a supplement containing principally 600 mg black tea extract (60 percent polyphenols, 20 percent caffeine) and 442 mg guarana extract (36 percent caffeine) or matching placebo. RESULTS: The RMR and systolic blood pressure (SBP) AUCs increased significantly (p less than 0.02 and p less than 0.01, respectively) in the herbal supplement group compared to placebo. The AUC increase in RMR over the two-hour test period was 77.19 kcal/24 hr2 +/- 120.10 kcal/24 hr2 with an average rise of 52.38 +/- 29.52 kcal/24 hrs. The AUC rise in SBP over two hours was 10.3 mm Hg/hr +/- 14 mm Hg/hr. The average rise in SBP over two hours was 3.7 mm Hg +/- 4.4 mm Hg. DISCUSSION: The herbal supplement increased metabolic rate without changing substrate oxidation. The rise in SBP was consistent with the amount of caffeine the supplement contained. PMID- 16366741 TI - Melatonin. Monograph. PMID- 16366742 TI - Indole-3-carbinol. Monograph. PMID- 16366743 TI - Looking for foes and friends: perceptual and emotional factors when finding a face in the crowd. AB - In a face-in-the-crowd setting, the authors examined visual search for photographically reproduced happy, angry, and fearful target faces among neutral distractor faces in 3 separate experiments. Contrary to the hypothesis, happy targets were consistently detected more quickly and accurately than angry and fearful targets, as were directed compared with averted targets. There was no consistent effect of social anxiety. A facial emotion recognition experiment suggested that the happy search advantage could be due to the ease of processing happy faces. In the final experiment with perceptually controlled schematic faces, the authors reported more effective detection of angry than happy faces. This angry advantage was most obvious for highly socially anxious individuals when their social fear was experimentally enhanced. PMID- 16366744 TI - Sensation seeking and the aversive motivational system. AB - Sensation seeking (SS) has traditionally been viewed as a phenomenon of the appetitive motivational system. The limited SS research exploring contributions from the aversive motivational system reveals greater anxious reactivity to dangerous activities among low sensation seekers. The present study extends this line of work by comparing levels of fear and anxiety during anticipation of predictable and unpredictable aversive stimuli across high- and low-SS groups. Low sensation seekers displayed greater fear-potentiated startle (FPS) to predictable aversive stimuli, and only those low on SS showed FPS and skin conductance response effects during experimental contexts in which aversive stimuli were delivered unpredictably. Findings implicate enhanced apprehensive anticipation among those low on SS as a potential deterrent for their participation in intense and threatening stimulus events. PMID- 16366745 TI - Emotional stimuli, divided attention, and memory. AB - The emotion-memory literature has shown that negative emotional arousal enhances memory. S. A. Christianson (1992) proposed that preattentive processing could account for this emotion-memory relationship. Two experiments were conducted to test Christianson's theory. In Experiment 1, participants were exposed to neutral and negative arousing slides. In Experiment 2, participants were exposed to neutral, negative arousing, and positive arousing slides. In both experiments, the aforementioned variable was factorially combined with a divided-attention or non-divided-attention condition. The authors predicted that, in contrast to the nondivided condition, dividing attention would adversely impact neutral and positive stimuli more than negative stimuli. The hypothesis was supported; participants recalled more high negative-arousal slides than positive or neutral slides when their attention was divided rather than nondivided. PMID- 16366746 TI - Abstract stimuli associated with threat through conditioning cannot be detected preattentively. AB - Studies of anxiety suggest that threat stimuli can be identified preattentively, but this conclusion is questionable because of possible low-level perceptual confounds. Two experiments used visual search tasks in which abstract shapes were conditioned to carry neutral or negative valence. Experiment 1 found generally faster responses to threat-associated abstract stimuli but no evidence that they were detected preattentively, irrespective of trait anxiety level. A similar pattern was found in Experiment 2, in which individuals high in snake or spider fear showed no evidence of preattentive detection of abstract stimuli associated with their feared object. In contrast, implicit behavioral measures showed significant effects of conditioning, demonstrating that targets associated with threat were negatively evaluated in these experiments. PMID- 16366747 TI - Measuring affective clarity indirectly: individual differences in response latencies of state. AB - This research investigated a new method to measure momentary affective clarity indirectly, which is based on latencies of responses to state affect items. Three studies revealed that this indirect measure of momentary clarity demonstrated high reliability and stability as well as convergent and predictive validity. The indirect measure was associated with dispositional clarity when the concept of clarity was activated before measuring response latencies (Studies 1 and 2) and was related to self-reports of momentary clarity (Study 3). Furthermore, Study 3 demonstrated that indirectly measured clarity decreased after an affectively complex film. Indirectly, but not directly, measured momentary clarity predicted a more positive affective state at the end of the study. This effect was mediated by affect regulation. PMID- 16366748 TI - Mood-congruent attentional bias in dysphoria: maintained attention to and impaired disengagement from negative information. AB - Attentional bias to negative information has been proposed to be a cognitive vulnerability factor for the development of depression. In 2 experiments, the authors examined mood-congruent attentional bias in dysphoria. In both experiments, dysphoric and nondysphoric participants performed an attentional task with negative, positive, and neutral word cues preceding a target. Targets appeared either at the same or at the opposite location of the cue. Overall, results indicate that dysphoric participants show maintained attention for negative words at longer stimulus presentations, which is probably caused by impaired attentional disengagement from negative words. Furthermore, nondysphoric participants maintain their attention more strongly to positive words. These results are discussed in relation to recent developments in the pathogenesis and treatment of depression. PMID- 16366749 TI - Defocused attention in depressed mood: evidence from source monitoring. AB - The authors suggest that depressed mood is associated with a defocused mode of attention, allowing irrelevant information to be noticed and processed more than in nondepressed states. Working on a source monitoring task, subclinically depressed college students selected with the Beck Depression Inventory (A. T. Beck, 1967; D. Kammer, 1983) had better memory for irrelevant stimulus aspects than nondepressed control students. However, depressed students' performance on the relevant stimulus aspects was unimpaired. These results are in conflict with a capacity reduction view of depressed mood and support the hypothesized altered, defocused mode, in which attentional resources are more evenly allocated across various aspects of the materials. The results are discussed within the framework of adaptive functions of emotional states. PMID- 16366750 TI - The worried mind: autonomic and prefrontal activation during worrying. AB - To study the psychophysiological correlates of worrying, the authors recorded heart rate, respiratory sinus arrhythmia (RSA), skin conductance level, and alpha electroencephalographic asymmetry in healthy males during baseline, relaxation, worry induction, and anticipation of an impromptu speech task. Compared with baseline, relaxation, and anticipation, worrying was associated with greater heart rate and lower RSA. Worrying was further characterized by higher skin conductance levels compared with baseline but lower levels than during anticipation. Finally, worrying was associated with relatively greater left frontal activity compared with anticipation. Trait public speaking anxiety was positively correlated with left frontal activity during worrying. These results support the notion that worrying is a unique emotional state that is different from fearful anticipation. PMID- 16366751 TI - Implicit self-attitudes predict spontaneous affect in daily life. AB - In 2 studies, the authors examined the degree to which implicit self-attitudes predicted people's spontaneous affective experiences in daily life. Across both studies, implicit attitudes toward the self (as measured by Implicit Association Tests) strongly predicted negative feeling states (as measured by computerized experience-sampling procedures), suggesting that implicit self-attitudes may be linked to changes in undifferentiated negative affect. Explicit attitudes toward the self generally did not account for these relations. Findings extend understanding of the factors that contribute to experienced affect and are the first to empirically link implicit self-attitudes with phenomenological affective experience in real-life settings over time. PMID- 16366752 TI - Mental imagery and emotion: a special relationship? AB - A special association between imagery and emotion is often assumed, despite little supporting evidence. In Experiment 1, participants imagined unpleasant events or listened to the same descriptions while thinking about their verbal meaning. Those in the imagery condition reported more anxiety and rated new descriptions as more emotional than did those in the verbal condition. In Experiment 2, 4 groups listened to either benign or unpleasant descriptions, again with imagery or verbal processing instructions. Anxiety again increased more after unpleasant (but not benign) imagery; however, emotionality ratings did not differ after a 10-min filler task. Results support the hypothesis of a special link between imagery and anxiety but leave open the question of whether this also applies to other emotions. PMID- 16366753 TI - Facial attractiveness is appraised in a glance. AB - Those who are physically attractive reap many benefits--from higher average wages to a wider variety of mate choices. Recent studies have investigated what constitutes beauty and how beauty affects explicit social judgments, but little is known about the perceptual or cognitive processing that is affected by aesthetic judgments of faces and why beauty affects our behavior. In this study, the authors show that beauty is perceived when information is minimized by masking or rapid presentation. Perceiving and processing beauty appear to require little attention and to bias subsequent cognitive processes. These facts may make beauty difficult to ignore, possibly leading to its importance in social evaluations. PMID- 16366754 TI - Influence of emotion on memory for temporal information. AB - Contextual information, such as color and spatial location, has been found to be better remembered for emotional than for neutral items. The current study examined whether the influence of emotion extends to memory for another fundamental feature of episodic memory: temporal information. Results from a list discrimination paradigm showed that (a) item memory was enhanced for both negative and positive pictures compared with neutral ones and was better for negative than for positive pictures and (b) temporal information was better remembered for negative than for positive and neutral pictures, whereas positive and neutral pictures did not differ from each other. These findings are discussed in relation to the processes involved in memory for temporal information. PMID- 16366755 TI - The use of ecological momentary assessment to test appraisal theories of emotion. AB - Although appraisal theories have received strong empirical support, there are methodological concerns about the research, including biased recall, heuristic responding, ethical issues, and weak and unrealistic induction of emotions in laboratories. To provide a more ecologically valid test of appraisal theories, the authors used ecological momentary assessment, in which the emotions and appraisals of Singaporean police officers were measured online over the course of an ordinary workday. The research focused on happiness. Support was obtained for predictions, demonstrating the generalizability of appraisal theories to a nonlaboratory setting and circumventing the shortcomings of previously used methodologies. Also, evidence was obtained that happiness was reported primarily in association with a specific combination of 3 relevant appraisals: high pleasantness, high perceived control, and low moral violation. PMID- 16366756 TI - Affective speech elicited with a computer game. AB - To determine the degree to which emotional changes in speech reflect factors other than arousal, such as valence, the authors used a computer game to induce natural emotional speech. Voice samples were elicited following game events that were either conducive or obstructive to the goal of winning and were accompanied by either pleasant or unpleasant sounds. Acoustic analysis of the speech recordings of 30 adolescents revealed that mean energy, fundamental-frequency level, utterance duration, and the proportion of an utterance that was voiced varied with goal conduciveness; spectral energy distribution depended on manipulations of pleasantness; and pitch dynamics depended on the interaction of pleasantness and goal conduciveness. The results suggest that a single arousal dimension does not adequately characterize a number of emotion-related vocal changes, lending weight to multidimensional theories of emotional response patterning. PMID- 16366757 TI - The influence of caffeine on nicotine's discriminative stimulus, subjective, and reinforcing effects. AB - Caffeine may acutely alter the discriminative stimulus and subjective effects of nicotine, perhaps explaining the association of coffee intake with smoking status. In this study, smokers were initially trained to discriminate 20 microg/kg nicotine by nasal spray from placebo (0). Then, generalization of nicotine discrimination was tested, using both 2- and 3-choice ("novel" option) procedures, across a range of doses (0-20 microg/kg) following pretreatment with 0, 2.5, and 5.0 mg/kg caffeine p.o. Nicotine reinforcement was assessed after the end of generalization testing using a choice procedure. Caffeine pretreatment did not alter nicotine discrimination and self-administration. Caffeine and nicotine influenced some subjective and cardiovascular responses, but there were no interaction effects except for diastolic blood pressure. These results do not support the notion that caffeine acutely alters nicotine's discriminative stimulus, subjective, or reinforcing effects. PMID- 16366759 TI - Predictors of outcome in LAAM, buprenorphine, and methadone treatment for opioid dependence. AB - This study examined (1) predictors of treatment outcome for opioid-dependent participants in a single-site controlled trial comparing methadone, buprenorphine, and LAAM treatments and (2) the extent to which various subpopulations of patients may have more successful outcomes with each medication. The relationships between patient demographics, drug use history, and psychological status and outcome measures of treatment retention, opiate use, and cocaine use were assessed. We believe this study to be the first to demonstrate that predictors of treatment success appear to be largely similar in LAAM, buprenorphine, and methadone treatment for opioid dependence. We did not find any factors that would strongly guide selection of one medication over others. PMID- 16366758 TI - Combined effects of venlafaxine, nicotine replacement, and brief counseling on smoking cessation. AB - In this study, 147 smokers were randomly assigned to receive either venlafaxine or placebo in conjunction with behavioral counseling (9 weekly sessions) and transdermal nicotine replacement therapy (22 mg/day). Patients began medication 2 weeks before quitting and continued for 18 weeks after quitting, with the daily dose titrated from 150 to 225 mg. in response to symptoms of negative affect and relapse. The results showed no main effect of treatment on abstinence. Post hoc analysis revealed that both at the end of treatment and at the 1-year follow-up smokers consuming less than a pack of cigarettes a day benefited from the addition of venlafaxine to the treatment regimen. Venlafaxine also reduced negative affect for all smokers for up to 6 weeks postcessation. The findings suggest that venlafaxine could have some role to play in the treatment of lighter smokers, in addition to the expected benefits of nicotine replacement therapy and behavioral counseling. PMID- 16366760 TI - Patterns of opioid and cocaine co-use: a descriptive study in a Canadian sample of untreated opioid-dependent individuals. AB - This study examined prevalence and patterns of co-use of opioids and cocaine in regular users of illicit opioids (N = 729) recruited from 5 Canadian cities. Fifty-seven percent (n = 417) reported having used both opioids and cocaine in the month and week preceding the interview; of these, 73% (n = 304) were able to identify a typical pattern of daily co-use. In a typical day, injectors of opioids and cocaine (n = 119) and injectors of opioids who inhaled cocaine (n = 111) showed stable opioid use but variable cocaine use, which peaked at 21 hr. Overall, 30% of the individuals used both drugs exclusively in a sequential fashion, 35% reported taking opioids and cocaine within the same hour, and 35% reported taking them together at the same time or mixing them. These findings indicate that different individuals display different patterns of opioids and cocaine co-use. PMID- 16366761 TI - Differences in impulsivity and risk-taking propensity between primary users of crack cocaine and primary users of heroin in a residential substance-use program. AB - Crack cocaine use is more associated with impulsivity and a propensity to take risks than heroin use, yet no studies have examined this relationship in the absence of acute drug effects. The current study examined impulsivity (using the Delay Discounting Task) and risk-taking propensity (using the Balloon Analogue Risk Task) across independent groups of primary crack cocaine users with minimal heroin use (n = 16) and primary heroin users with minimal crack cocaine use (n = 11) in residential treatment, with all participants drug abstinent during participation. Crack cocaine users evidenced greater levels of impulsivity and risk-taking propensity, with only the difference in impulsivity persisting after controlling for age and gender. These data hold potential theoretical importance in understanding differences between crack cocaine and heroin users, as the findings cannot be attributed solely to acute pharmacological drug effects. PMID- 16366762 TI - Effects of repeated-dose isradipine on the abuse liability of cocaine. AB - Despite preclinical studies suggesting that isradipine may antagonize the abuse liability of cocaine, pretreatment with sustained-release isradipine did not reduce euphoric mood in cocaine-using volunteers. This double-blind, within subject, crossover laboratory study determined whether maximal dose-loading with isradipine could antagonize effects of cocaine in 12 cocaine-dependent research volunteers administered intravenous cocaine doses (0, 0.325, and 0.65 mg/kg) on different days after 5 days of treatment with isradipine or placebo. Isradipine dose was 30 mg sustained release nightly plus 15 mg immediate release 2 hr before cocaine infusion. Cocaine produced dose-related increases in cocaine's subjective effects and a behavioral measure of reinforcement. Isradipine enhanced, rather than antagonized, subjective effects, indicating that isradipine does not antagonize cocaine's abuse liability in dependent research volunteers. PMID- 16366764 TI - Drugs, memory, and metamemory: a dose-effect study with lorazepam and scopolamine. AB - This experiment was designed to use the graded dose-related amnesia produced by the benzodiazepine lorazepam (1.0, 2.0 mg/70 kg, oral) and the anticholinergic scopolamine (0.3, 0.6 mg/70 kg, subcutaneous) as a tool to explore the cognitive and neurochemical mechanisms underlying metamemory in the judgment of learning paradigm, with a placebo-controlled independent groups design in healthy volunteers (n = 12/group). Results provide evidence for a pharmacological dissociation between effects on memory versus metamemory (relative accuracy of item-by-item monitoring) across a range of levels of memory performance and suggest that the drugs selectively impair those aspects of metamnemonic monitoring that require participants' awareness of their overall current state of functioning (absolute accuracy of prospective item-by-item monitoring, prospective global monitoring) but not those that rely solely on assessment of individual item characteristics (relative accuracy of item-by-item monitoring). PMID- 16366763 TI - Cocaine improves inhibitory control in a human model of response conflict. AB - The present study was designed to test the acute effects of cocaine on behavioral control in the presence and absence of motivational conflict. Adults (N = 14) with a history of stimulant use received oral cocaine hydrogen chloride (0, 100, 200, and 300 mg) and performed a cue-dependent go/no-go task to measure inhibitory and activational mechanisms of behavioral control either with or without motivated conflict between the inhibition and the activation of responses. Cocaine improved response inhibition in both conflict conditions, as evident by a decrease in inhibitory failures following active doses. The current study provides a useful model to investigate the effects of other drugs reported to have performance-enhancing effects. PMID- 16366765 TI - Reinforcing effects of diazepam under anxiogenic conditions in individuals with social anxiety. AB - Diazepam (DZ) reinforcement was tested under anxiogenic (public speaking) and neutral (computer task) conditions. Individuals with social anxiety disorder (n = 11) and healthy controls (n = 11) participated in two 5-session phases. Each phase used a standard choice procedure (2 sample, 3 choice sessions) comparing 10 mg DZ and placebo. During the public speaking condition, DZ preference was greater among the participants with social anxiety compared with controls (81.8% vs. 36.4%; p < .05). Participants with social anxiety also gave DZ significantly higher crossover values on the multiple choice procedure under the speech condition compared with the computer condition. Subjective effects indicated that DZ reduced anxiety when levels were elevated during the speech in socially anxious participants. These results suggest that DZ reinforcement may occur under conditions of heightened anxiety by bestowing therapeutic efficacy. PMID- 16366766 TI - The relation between social problems and substance use in adolescent boys: an investigation of potential moderators. AB - This study investigated the relation between social problems and adolescent male substance use in the context of other potential moderating variables. Two hundred eighty adolescent boys completed measures of social problems, cognitive distortions, constructive thinking, affiliation with delinquent peers, and multiple aspects of substance use. Results revealed that social problems were not directly related to most substance use variables. Cognitive distortions moderated the relation between social problems and substance use in social situations, such that the relation was significant only at high levels of cognitive distortions. Constructive thinking and affiliation with delinquent peers were both related to substance use, but neither served a moderating function. It is concluded that the widely accepted relation between social difficulties and adolescent male substance use may be better understood in the context of cognitive variables. PMID- 16366767 TI - Novelty seeking and drug use: contribution of an animal model. AB - Although sensation seeking or novelty seeking is a reliable predictor of drug use in humans, individual differences in free-choice novelty seeking in animal models have generally failed to predict drug use. In the current article, hierarchical multiple regression analyses were used on data collected from a large sample of rats. Rats were screened on measures of inescapable and free-choice novelty tests and then were trained to lever press for sucrose or intravenous amphetamine. Although scores from the inescapable novelty test weakly predicted responding for amphetamine, the addition of free-choice novelty preference scores into the regression analyses significantly improved the predictive models. These results indicate that, similar to evidence in humans, individual differences in novelty seeking may be able to predict drug use in rats. PMID- 16366768 TI - Incentive program decreases no-shows in nontreatment substance abuse research. AB - This study examined the use of a low-cost incentive program to decrease the rate of unanticipated no-shows in a nontreatment study of the genetics of substance dependence. Low-cost retail items (such as calling cards or gift certificates) were offered contingent on attendance at the first scheduled research appointment. Although the intervention did not result in an increase in the rate of attendance at appointments, it reduced the likelihood of unanticipated no shows by almost 50%. This reduction was accompanied by a significant increase in advance cancellations. Despite limitations due to the study's A-B design, this research demonstrates that it may be possible to use modest incentives to increase the efficiency of research enrollment and increase service provision for substance abusers. PMID- 16366770 TI - Acoustic mechanisms of note-type perception in black-capped chickadee (Poecile atricapillus) calls. AB - Acoustic communication in black-capped chickadees (Poecile atricapillus) has been studied intensively, the "chick-a-dee" call being among the most well described. This call consists of 4 note types; chickadees perceive these notes as open-ended categories and do so in a continuous manner, with As more similar to Bs and Bs more similar to Cs. Acoustic features contributing to the note-type differentiation are unknown. Recent analyses suggested that certain acoustic features may play a role in note-type classification. Here, the authors tested black-capped chickadees in an operant-conditioning paradigm to determine which features were controlling note-type perception. The results suggest that the note pitch and the frequency modulation in the initial portion of the note control the perception of note types. PMID- 16366769 TI - Factors influencing the prevalence and handedness for throwing in captive chimpanzees (Pan troglodytes). AB - Humans throw right-handed, and it has been suggested that the neurophysiological demands of aimed throwing may have served as a precursor to the evolution of left hemisphere specialization for linguistic functions. Although there are descriptions of throwing by wild and captive chimpanzees (Pan troglodytes), systematic observations of aimed throwing and handedness have not been reported. In this article, evidence of population-level right-handedness for throwing is reported in 2 samples of captive chimpanzees. It is further reported that right handed throwing is more pronounced than other measures of handedness in captive chimpanzees. The implications of these findings are discussed in the context of theories relating throwing to the evolution of lateralization for language functions. PMID- 16366771 TI - Life history and social learning: megapode chicks fail to acquire feeding preferences from conspecifics. AB - Australian brush-turkeys, Alectura lathami, are birds with an unusual life history: The young receive no parental care and first encounter conspecifics at an unpredictable age. Brush-turkey chicks that were 3-4 days old were presented with a robot model that appeared to feed from a distinctively colored dish. In control training trials, chicks saw a robot standing next to a different dish and scanning from side to side. Chicks expressed a strong tendency to feed from dishes of the type indicated by the pecking robot, but this effect proved ephemeral. Brush-turkeys hence appear to show no social learning under conditions that inculcate stable preferences in other galliforms such as chickens, suggesting that life history plays an important role in the evolution of learning. PMID- 16366772 TI - Further evidence that mice learn a win-shift but not a win-stay contingency under water-escape motivation. AB - Previous research (C. Locurto, C. Emidy, & S. Hannan, 2002) indicated that mice quickly learned a water-escape task under a win-shift contingency but did not exceed chance-level performance under a win-stay contingency. We examined the robustness of this conclusion in two experiments by varying procedural and temporal aspects of that earlier experiment. Results of both experiments indicated that the preference for win-shift learning in mice under water-escape motivation could not be attributed to procedural or design features of that earlier study and were independent of the influence of intertrial interval, normally a variable that produces strong effects on learning. In neither experiment did subjects exposed to a win-stay contingency perform at above-chance levels. PMID- 16366773 TI - How prosimian primates represent tools: experiments with two lemur species (Eulemur fulvus and Lemur catta). AB - The authors examined how 2 lemur species (Eulemur fulvus and Lemur catta) reason about tools. Experiment 1 allowed subjects to use 1 of 2 canes to retrieve an inaccessible food reward. Lemurs learned to solve this problem as quickly as other primates. Experiment 2 then presented subjects with novel tools differing from the originals along one featural dimension. Subjects attended more to tools' sizes than to their colors and made no distinction between tools' shapes and textures. Experiments 3 and 4 presented problems in which some of the tools' orientations had to be modified relative to the food. Subjects performed well on these problems, sometimes modifying the position of the tool. These results are discussed in light of the performance of other primates on this task. PMID- 16366774 TI - Recent use of signs by chimpanzees (Pan Troglodytes) in interactions with humans. AB - In light of the controversy about the linguistic properties of chimpanzee signing behavior, the recent sign use of 5 chimpanzees (Pan troglodytes) with long histories of sign use was analyzed while they interacted with longtime human companions. Four corpora from 1992 to 1999 consisting of 3,448 sign utterances were examined. The chimpanzees predominantly used object and action signs. There was no evidence for semantic or syntactic structure in combinations of signs. Longer combinations showed repetition and stringing of object and action signs. The chimpanzees mostly signed with an acquisitive motivation. Requests for objects and actions were the predominant communicative intentions of the sign utterances, though naming and answering also occurred. This recent sign use shows multiple differences with (early) human language. PMID- 16366775 TI - Housing pregnant mice (Mus musculus) in small groups facilitates the development of odor-based homing in offspring. AB - Infant mice (Mus musculus) born to dams housed in isolation throughout pregnancy (IsoPreg) begin differentially approaching homenest bedding over clean bedding on Postnatal Day 6. Offspring of dams housed with 2 other potentially pregnant conspecifics (SocPreg) display such homing behavior on Day 4. Earlier onset of homing reflects facilitated olfactory responsiveness in SocPreg pups, rather than qualitative or quantitative differences in IsoPreg versus SocPreg nest odors, body growth, or motoric capabilities. Exposing pregnant IsoPreg dams to SocPreg bedding also accelerated homing onset in the offspring, though not to the same extent as the full social context. Thus, it appears that the facilitation of homing is mediated through the pregnant dam by a combination of chemical cues and other social stimuli. PMID- 16366776 TI - Reach and posture hand preferences during arboreal feeding in Sifakas (Propithecus sp.): a test of the postural origins theory of behavioral lateralization. AB - Sixteen sifakas (11 Propithecus verreauxi coquereli, 2 Propithecus verreauxi verreauxi form majori, and 3 Propithecus tattersalli) were videotaped as they fed on leaves in an arboreal context. The hand used to feed and the hand used to maintain postural stability was coded. For each subject, the lateral bias of the hand used to feed was opposite the hand used in postural support. Seven sifakas displayed no bias for feeding or posture-related hand use, 7 sifakas displayed significant feeding-related reach preferences for pulling branches to the mouth (5 left- and 2 right-hand preferences), and 9 sifakas exhibited significant hand preferences for postural support (2 left-, 7 right-hand preferent). Although these data do not strongly support the postural origins theory of behavioral lateralization, the modal preference pattern for sifakas that displayed significant hand preferences for posture and feeding involve a left bias for feeding and a right bias for postural support. s reserved). PMID- 16366777 TI - Female Japanese quail (Coturnix japonica) mated with males that harassed them are unlikely to lay fertilized eggs. AB - Results of previous studies of courtship and mating in Japanese quail (Coturnix japonica) suggest that females avoid conspecific males because, while courting and mating, males engage in behaviors that are potentially injurious to females. However, prior experiments provided no direct evidence that females avoided harassing males. Here the authors show that a female quail choosing between a previous sex partner and an unfamiliar male avoids the former if he engaged in relatively many potentially injurious acts while courting and mating, (Experiments 1 and 2) and that males behaving aggressively toward mates are less likely than are gentler males to fertilize the females' eggs (Experiment 3). Male sexual harassment appears to be a tactic both aversive to female quail and relatively ineffective in fertilizing them. PMID- 16366778 TI - Snake mating systems, behavior, and evolution: the revisionary implications of recent findings. AB - Sexual selection and mating systems profoundly influence the behavior and psychology of animals. Using their own studies of green anacondas (Eunectes murinus) and reviewing other recent studies, the authors conclude that incomplete data derived from a few well-studied snake species have led to general acceptance of polygyny as the dominant mating system in snakes. New data on behavior, paternity, and life history in a diverse taxonomic array of snakes support the view that polyandry is not only common in snakes but may have been the ancestral mating system. This interpretation helps to explain many seemingly paradoxical behavioral differences between lizards and snakes, such as the lack of territorial systems in most snakes and their frequent female-biased sexual size dimorphism. PMID- 16366779 TI - Goldfish (Carassius auratus) matching geometric and featural cues: a reinterpretation of some of the data of Vargas, Lopez, Salas, and Thinus-Blanc (2004). AB - Vargas, Lopez, Salas, and Thinus-Blanc showed that goldfish (Carassius auratus) can use both geometric and featural cues in relocating a target corner in a rectangular enclosure. When featural cues (arrangement of striped walls) were put in conflict with geometric cues, results differed according to target location during training. Vargas, Lopez, et al. explained the results of their cue conflict in terms of 2 different strategies: mapping and cue guidance. I provide an alternative, more parsimonious interpretation in which the same strategy of attempting to match as many cues as possible applies to both cases. PMID- 16366780 TI - Different ways of encoding geometric information by goldfish (Carassius auratus). AB - On the basis of results of a probe trial in 2 different experiments, K. Cheng (2005) has proposed a common mechanism for orientation in fish trained in both a map-like or relational procedure and a directly cued procedure. However, K. Cheng's model is inconsistent with previous results of goldfish (Carassius auratus) trained in these 2 tasks. Given that K. Cheng's proposal assumes that fish choose the goal by using a matching strategy in which they try to match as many properties as possible, including geometric and featural properties, future research is necessary to clarify what properties of the environmental space are codified and used for navigation. PMID- 16366781 TI - Attention to combined attention in new world monkeys (Cebus apella, Saimiri sciureus). AB - Co-orientation by capuchin (Cebus apella) and squirrel (Saimiri sciureus) monkeys in response to familiar humans abruptly switching the direction of their visual attention was recorded. Co-orientation occurred more frequently overall in capuchins than squirrel monkeys. Capuchins showed a tendency to habituate within trials involving consecutive attention switches performed by 2 different people, whereas squirrel monkeys co-oriented more when the 2nd attention switch was by a 2nd actor. These results suggest variable attention-processing abilities in New World monkeys, including differences in summation of attention by others. PMID- 16366782 TI - On the role of object representations in substitution masking. AB - Three experiments were conducted to investigate the role of object representations in object substitution masking (OSM). OSM occurs when a very sparse mask is presented simultaneously with a target stimulus and the target offsets first, leaving the mask to linger in the display for some time. Results confirm earlier claims that there is an isolatable object-level component to OSM and indicate that a target can be protected from OSM if, prior to offsetting, it can be represented as a distinct object from the mask. When the mask was presented as sliding past the target (Experiment 1), as jiggling independently of the target (Experiment 2), or in a different color from the target (Experiment 3), OSM was reduced or eliminated. This suggests that OSM reflects basic updating processes that allow the perception of continuity of object identity over change. PMID- 16366783 TI - Categorical and coordinate relations in faces, or Fechner's law and face space instead? AB - E. E. Cooper and T. J. Wojan (2000) applied the categorical-coordinate relations distinction to faces on the basis of a finding that two-eyes-up versus one-eye-up distortions had opposite effects in between-class (face normality) and within class (face identity) tasks. However, Cooper and Wojan failed to match amount of metric change between their 2 deviation types and tested only 1 deviation level. In the present study, eyeheight was shifted (e.g., both eyes up or both eyes down vs. one eye up and one eye down) parametrically (11 levels) and normality and identity ratings obtained. There was no evidence of categorical changes in perception where these would have been predicted by Cooper and Wojan's theory. In all cases, the relationship between physical and perceived distortion followed Fechner's law. Differences across distortion types in Fechner threshold (the minimum deviation altering perceived normality or identity) are explained in terms of the variability associated with different dimensions in face space. PMID- 16366784 TI - Evidence of separable spatial representations in a virtual navigation task. AB - Three experiments investigated spatial orientation in a virtual navigation task. Subjects had to adjust a homing vector indicating their end position relative to the origin of the path. It was demonstrated that sparse visual flow was sufficient for accurate path integration. Moreover, subjects were found to prefer a distinct egocentric or allocentric reference frame to solve the task. "Turners" reacted as if they had taken on the new orientation during turns of the path by mentally rotating their sagittal axis (egocentric frame). "Nonturners," by contrast, tracked the new orientation without adopting it (allocentric frame). When instructed to use their nonpreferred reference frame, both groups displayed no decline in response accuracy relative to their preferred frame; even when presented with reaction formats based on either ego or allocentric coordinates, with format unpredictable on a trial, both groups responded highly accurately. These findings support the assumption of coexisting spatial representations during navigation. PMID- 16366785 TI - Inhibition of return to an occluded object depends on expectation. AB - Inhibition of return (IOR) is indexed by slower reaction times to targets presented at previously attended locations or objects. If a moving object is occluded, some studies find IOR, others do not. Four experiments examined whether this inconsistency hinges on the observer's expectation as to whether the object continues to exist at the end of its motion sequence. Results showed that observer expectation is a powerful determining factor: IOR occurs only if the observer expects the object to continue to exist. In contrast, if the object is not occluded, IOR occurs only if the object remains on view immediately before the target is presented. It was concluded that 2 factors, object continuity and observer expectation, mediate both location- and object-based IOR. PMID- 16366786 TI - How two share a task: corepresenting stimulus-response mappings. AB - Previous research has shown that individuals unintentionally adjust their behavior to others by mimicking others' actions and by synchronizing their actions with others. This study investigated whether individuals form a representation of a coactor's task when the context does not require interpersonal coordination. Pairs of participants performed a reaction time (RT) task alongside each other, responding to 2 different dimensions of the same stimulus. Results showed that each actor's performance was influenced by the other's task. RTs on trials that required a response from both participants were slowed compared with trials that required only a response from 1 actor. Similar results were observed when each participant knew the other's task but could not observe the other's actions. These findings provide evidence that shared task representations are formed in social settings that do not require interpersonal coordination and emerge as a consequence of how a social situation is conceptualized. PMID- 16366787 TI - Gender recognition from point-light walkers. AB - Point-light displays of human gait provide information sufficient to recognize the gender of a walker and are taken as evidence of the exquisite tuning of the visual system to biological motion. The authors revisit this topic with the goals of quantifying human efficiency at gender recognition. To achieve this, the authors first derive an ideal observer for gender recognition on the basis of center of moment (J. E. Cutting, D. R. Proffitt, & L. T. Kozlowski, 1978) and, with the use of anthropometric data from various populations, show optimal recognition of approximately 79% correct. Next, they perform a meta-analysis of 21 experiments examining gender recognition, obtaining accuracies of 66% correct for a side view and 71% for other views. Finally, results of the meta-analysis and the ideal observer are combined to obtain estimates of human efficiency at gender recognition of 26% for the side view and 47% for other views. PMID- 16366788 TI - The developmental roots of the speed-accuracy trade-off. AB - The speed of adult reaching movements is lawfully related to the distance of the reach and the size of the target. The authors had 7-, 9-, and 11-month-old infants reach for small and large targets to investigate a possible relation between the emergence of this speed-accuracy trade-off and the improvements in infants' ability to pick up tiny objects. By 7 months of age, infants slowed down their reaches for smaller objects. The authors concluded that it was not the ability to use a precision grip that facilitated the speed-accuracy trade-off but rather the other way around. The slowing down toward the end of the movement might set the conditions for the development of fine distal control of the hand. PMID- 16366789 TI - Visual search asymmetry with uncertain targets. AB - The underlying mechanism of search asymmetry is still unknown. Many computational models postulate top-down selection of target-defining features as a crucial factor. This feature selection account implies, and other theories implicitly assume, that predefined target identity is necessary for search asymmetry. The authors tested the validity of the feature selection account using a singleton search task without a predefined target. Participants conducted a target-defined and a singleton search task with a circle (O) and a circle with a vertical bar (Q). Search asymmetry was observed in both tasks with almost identical magnitude. The results were not due to trial-by-trial feature selection, because search asymmetry persisted even when the target was completely unpredictable. Asymmetry in the singleton search was also observed with more complex stimuli, Kanji characters. These results suggest that feature selection is not necessary for search asymmetry, and they impose important constraints on current visual search theories. PMID- 16366790 TI - Boundary extension: the role of magnification, object size, context, and binocular information. AB - Boundary extension is a tendency to remember close-up scenes as if they extended beyond the occluding boundaries. The authors explored the contributing factors using brief retention intervals and computer-generated images. Boundary extension turns out to be more complex than previously thought and is not linked to the effects of image magnification and field-of-view changes. Although this is consistent with the idea that boundary extension is the product of the activation of a mental schema that provides information of what is likely to exist outside the picture boundaries, the authors also found that properties of the object at the center of the picture can affect boundary extension independently of the information at the boundaries. In a test of boundary extension using stereograms, the effect does not seem to depend on amount of perceived depth, suggesting a weaker link to perception of space than previously hypothesized. PMID- 16366791 TI - Representation of lexical form: evidence from studies of sublexical ambiguity. AB - The authors examined the role of intermediate, sublexical representations in spoken word perception. In particular, they tested whether flaps, which are neutralized allophones of intervocalic /t/s and /d/s, map onto their underlying phonemic counterparts. In 2 shadowing tasks, the authors found that flaps primed their carefully articulated counterparts, and vice versa. Because none of the flapped stimuli were lexically ambiguous (e.g., between rater and raider), these results provide evidence that such priming is sublexically mediated. Therefore, the current study provides further insights into when underlying form-based representations are activated during spoken word processing. In particular, the authors argue that phonological ambiguity, inherent in their flapped stimuli, is one of the conditions leading to the activation of underlying representations. PMID- 16366792 TI - Dissociations in perceptual learning revealed by adult age differences in adaptation to time-compressed speech. AB - When presented with several time-compressed sentences, young adults' performance improves with practice. Such adaptation has not been studied in older adults. To study age-related changes in perceptual learning, the authors tested young and older adults' ability to adapt to degraded speech. First, the authors showed that older adults, when equated for starting accuracy with young adults, adapted at a rate and magnitude comparable to young adults. However, unlike young adults, older adults failed to transfer this learning to a different speech rate and did not show additional benefit when practice exceeded 20 sentences. Listeners did not adapt to speech degraded by noise, indicating that adaptation to time compressed speech was not attributable to task familiarity. Finally, both young and older adults adapted to spectrally shifted noise-vocoded speech. The authors conclude that initial perceptual learning is comparable in young and older adults but maintenance and transfer of this learning decline with age. PMID- 16366793 TI - Auditory feedback in music performance: the role of melodic structure and musical skill. AB - Five experiments explored whether fluency in musical sequence production relies on matches between the contents of auditory feedback and the planned outcomes of actions. Participants performed short melodies from memory on a keyboard while musical pitches that sounded in synchrony with each keypress (feedback contents) were altered. Results indicated that altering pitch contents can disrupt production, but only when altered pitches form a sequence that is structurally similar to the planned sequence. These experiments also addressed the role of musical skill: Experiments 1 and 3 included trained pianists; other experiments included participants with little or no musical training. Results were similar across both groups with respect to the disruptive effects of auditory feedback manipulations. These results support the idea that a common hierarchical representation guides sequences of actions and the perception of event sequences and that this coordination is not acquired from learned associations formed by musical skill acquisition. PMID- 16366794 TI - Preview search and contextual cuing. AB - We investigated the effect of contextual cuing (M. M. Chun & Y. Jiang, 1998) within the preview paradigm (D. G. Watson & G. W. Humphreys, 1997). Contextual cuing was shown with a 10-item letter search but not with more crowded 20-item displays. However, contextual learning did occur in a preview procedure in which 10 preview items were followed by 10 new items. Repeating the new items alone did not generate contextual learning, but repeating the preview items alone did, as long as they had a consistent spatial relation with the target. This was not merely due to the onset of the preview items being associated with the target location. No learning effect took place with a preview of homogeneous items that competed less for selection with new stimuli. The results provide evidence for old items being processed in preview search and providing a context for subsequent search of new items. PMID- 16366795 TI - Illusion effects on grasping are temporally constant not dynamic. AB - The authors tested whether the effects of the Ebbinghaus illusion on grasping are corrected during late phases of the movement. Surprisingly, the grasp aperture was corrected neither under no-vision (N = 52) nor under full-vision (N = 48) conditions. The authors show that previous reports of a correction (e.g., S. Glover & P. Dixon, 2002a) are due to 2 artifacts: (a) inclusion of time points at which the target object was already touched and (b) erroneous statistics. This removes the central evidence on which S. Glover and P. Dixon's (2001a) planning control model of action is based. In addition, the authors' results can help to refine more classic notions of motor control (e.g., R. Woodworth, 1899). In consequence, the authors reject S. Glover and P. Dixon's (2001a) planning-control model but not classic online-control theories. PMID- 16366796 TI - The role of feedback information for calibration and attunement in perceiving length by dynamic touch. AB - Two processes have been hypothesized to underlie improvement in perception: attunement and calibration. These processes were examined in a dynamic touch paradigm in which participants were asked to report the lengths of unseen, wielded rods differing in length, diameter, and material. Two experiments addressed whether feedback informs about the need for reattunement and recalibration. Feedback indicating actual length induced both recalibration and reattunement. Recalibration did not occur when feedback indicated only whether 2 rods were of the same length or of different lengths. Such feedback, however, did induce reattunement. These results suggest that attunement and calibration are dissociable processes and that feedback informs which is needed. The observed change in variable use has implications also for research on what mechanical variables underlie length perception by dynamic touch. PMID- 16366797 TI - Reorienting in images of a three-dimensional environment. AB - Adult humans searched for a hidden goal in images depicting 3-dimensional rooms. Images contained either featural cues, geometric cues, or both, which could be used to determine the correct location of the goal. In Experiment 1, participants learned to use featural and geometric information equally well. However, men and women showed significant differences in their use of distant featural cues and the spontaneous encoding of geometric information when trained with features present. Transformation tests showed that participants could use either the color or the shape of the features independently to locate the goal. Experiment 2 showed that participants could use either configural or surface geometry when searching for the goal. However, their weighing of these geometric cues was dependent on initial training experience. PMID- 16366798 TI - Emotional devaluation of distracting patterns and faces: a consequence of attentional inhibition during visual search? AB - Visual search has been studied extensively, yet little is known about how its constituent processes affect subsequent emotional evaluation of searched-for and searched-through items. In 3 experiments, the authors asked observers to locate a colored pattern or tinted face in an array of other patterns or faces. Shortly thereafter, either the target or a distractor was rated on an emotional scale (patterns, cheerfulness; faces, trustworthiness). In general, distractors were rated more negatively than targets. Moreover, distractors presented near the target during search were rated significantly more negatively than those presented far from the target. Target-distractor proximity affected distractor ratings following both simple-feature and difficult-conjunction search, even when items appeared at different locations during evaluation than during search and when faces previously tinted during search were presented in grayscale at evaluation. An attentional inhibition account is offered to explain these effects of attention on emotional evaluation. PMID- 16366799 TI - Task demands control acquisition and storage of visual information. AB - Attention and working memory limitations set strict limits on visual representations, yet researchers have little appreciation of how these limits constrain the acquisition of information in ongoing visually guided behavior. Subjects performed a brick sorting task in a virtual environment. A change was made to 1 of the features of the brick being held on about 10% of trials. Rates of change detection for feature changes were generally low and depended on the pick-up and put-down relevance of the feature to the sorting task. Subjects' sorting decision suggests that changes may be missed because of a failure to update the changed feature. The authors also explore how hand and eye behavior are coordinated for strategic acquisition and storage of visual information throughout the task. PMID- 16366800 TI - Hyperspecificity in visual implicit learning: learning of spatial layout is contingent on item identity. AB - Humans conduct visual search faster when the same display is presented for a 2nd time, showing implicit learning of repeated displays. This study examines whether learning of a spatial layout transfers to other layouts that are occupied by items of new shapes or colors. The authors show that spatial context learning is sometimes contingent on item identity. For example, when the training session included some trials with black items and other trials with white items, learning of the spatial layout became specific to the trained color--no transfer was seen when items were in a new color during testing. However, when the training session included only trials in black (or white), learning transferred to displays with a new color. Similar results held when items changed shapes after training. The authors conclude that implicit visual learning is sensitive to trial context and that spatial context learning can be identity contingent. PMID- 16366801 TI - The efficiency of feature-based subitization and counting. AB - The enumeration of small numbers of objects (approximately 4) proceeds rapidly, accurately, and with little effort via a process termed subitization. Four experiments examined whether it was possible to subitize the number of features rather than objects present in a display. Overall, the findings showed that when features are presented randomly and are uncorrelated with object numerosity, efficient enumeration is not possible. This suggests that the visual system does not have parallel access to multiple feature maps and that subitization processes operate exclusively on representations coding the locations of objects. The data are discussed with respect to theories of visual enumeration and search. PMID- 16366802 TI - Delayed attentional engagement in the attentional blink. AB - Observers often miss the 2nd of 2 visual targets (first target [T1] and second target [T2]) when these targets are presented closely in time; the attentional blink (AB). The authors hypothesized that the AB occurs because the attentional response to T2 is delayed by T1 processing, causing T2 to lose a competition for attention to the item that follows it. The authors investigated this hypothesis by determining whether the AB is attenuated when T2 is precued. The results from 4 experiments showed that the duration and magnitude of the AB were substantially reduced when T2 was precued. The observed improvement in T2 report did not occur at the expense of T1 report, suggesting that processing of T1 was already completed or was at least protected when the cue was presented. The authors conclude that, during the AB, there is a delay between detection and the selection of target candidates for consolidation in short-term memory. PMID- 16366803 TI - Perception of objects in natural scenes: is it really attention free? AB - Studies have suggested attention-free semantic processing of natural scenes in which concurrent tasks leave category detection unimpaired (e.g., F. Li, R. VanRullen, C. Koch, & P. Perona, 2002). Could this ability reflect detection of disjunctive feature sets rather than high-level binding? Participants detected an animal target in a rapid serial visual presentation (RSVP) sequence and then reported its identity and location. They frequently failed to identify or to localize targets that they had correctly detected, suggesting that detection was based only on partial processing. Detection of targets was considerably worse in sequences that also contained humans, presumably because of shared features. When 2 targets were presented in RSVP, a prolonged attentional blink appeared that was almost eliminated when both targets were detected without being identified. The results suggest rapid feature analysis mediating detection, followed by attention demanding binding for identification and localization. PMID- 16366804 TI - Selective attention and the perception of an attended nontarget object. AB - Although many theories of attention assume that attending to an object results in the processing of all its feature dimensions, there has been no direct evidence that the irrelevant dimensions of an attended nontarget object are encoded. This article explores factors that modulate such processing. In 6 experiments, participants made a speeded response to a probe preceded by a prime that varied in 2 dimensions. Their reaction times to the probe were influenced by the response compatibility between the relevant and irrelevant dimensions of the prime. Furthermore, the effect was observed only when attention was directed to a nonlocation object feature and when participants' reaction times were relatively long. These results suggest that the effect of attention on a nontarget object is more complex than was previously understood. PMID- 16366805 TI - Updating target location at the end of an orienting saccade affects the characteristics of simple point-to-point movements. AB - Six results are reported. (a) Reaching accuracy increases when visual capture of the target is allowed (e.g., target on vs. target off at saccade onset). (b) Whatever the visual condition, trajectories diverge only after peak acceleration, suggesting that accuracy is improved through feedback mechanisms. (c) Feedback corrections are smoothly implemented, causing the corrected and uncorrected velocity profiles to exhibit similar shapes. (d) Initial kinematics poorly predict final accuracy whatever the condition, indicating that target capture is not the only critical input for feedback control. (e) Hand and eye final variability are unrelated, suggesting that gaze direction is not a target signal for arm control. (f) Extent errors are corrected without modification of movement straightness; direction errors cause path curvature to increase. Together these data show that movements with straight paths and bell-shaped velocity profiles are not necessarily ballistic. PMID- 16366807 TI - Changes in drug use during young adulthood: the effects of parent alcoholism and transition into marriage. AB - The present study examined the changes in drug use during the transition from emerging adulthood into young adulthood among a community sample of children of alcoholics (COAs) and demographically matched non-COAs. Consistent with national data, the non-COAs significantly decreased their drug use during this time, but the COAs did not significantly decrease their use. On the basis of role compatibility theory, the authors next examined whether marital status mediated or moderated this difference between COAs and non-COAs in linear drug use growth trajectories. In support of mediation, the authors found that COAs were significantly less likely than non-COAs to be currently married and that, for male participants, marriage was significantly associated with greater decreases in drug use during the mid to late 20s. PMID- 16366806 TI - Similarities and differences of longitudinal phenotypes across alternate indices of alcohol involvement: a methodologic comparison of trajectory approaches. AB - Extant developmental research distinguishing young adults who moderate versus persist in alcohol consumption has not consistently evaluated the domain of alcohol involvement being modeled, making it difficult to compare findings across studies. In the present study, the authors characterized the developmental course of 5 indices of alcohol involvement using a prospective (6-wave) sample of 377 young adults (Year 1 age = 18.52 years; 55% female; 51% with family history of alcoholism) over 11 years. Growth mixture models were applied to each measure. Despite similarity in trajectory shape, predicted prevalences varied, and the consistency of trajectory classifications across alternate indices revealed low agreement. Correlates of drinking course, however, were somewhat robust across alcohol index. The finding that trajectories are conditional on the specific indices used suggests that it may be hazardous to generalize across alternate indices of alcohol involvement. PMID- 16366808 TI - Brief relationship therapy for alcoholism: a randomized clinical trial examining clinical efficacy and cost-effectiveness. AB - The purpose of this study was to examine the clinical efficacy and cost effectiveness of brief relationship therapy (BRT), a shortened version of standard behavioral couples therapy (S-BCT), with alcoholic male patients (N = 100) and their nonsubstance-abusing female partners. Participants were randomly assigned to 1 of 4 treatment conditions: (a) BRT, (b) S-BCT, (c) individual-based treatment (IBT), or (d) psychoeducational attention control treatment (PACT). Equivalency testing revealed that, compared with those assigned to S-BCT, participants who were randomly assigned to BRT had equivalent posttreatment and 12-month follow-up heavy drinking outcomes. Moreover, at 12-month follow-up, heavy drinking and dyadic adjustment outcomes for patients who received BRT were superior to those of patients who received IBT or PACT. BRT was significantly more cost effective than the S-BCT, IBT, or PACT. PMID- 16366809 TI - Randomized study of combined universal family and school preventive interventions: patterns of long-term effects on initiation, regular use, and weekly drunkenness. AB - This study reports findings on a combined family and school-based competency training intervention from an in-school assessment 2.5 years past baseline, as a follow-up to an earlier study of substance initiation. Increased rates of observed alcohol use and an additional wave of data allowed evaluation of regular alcohol use and weekly drunkenness, with both point-in-time and growth curve analyses. Thirty-six rural schools were randomly assigned to (a) a combined family and school intervention condition, (b) a school-only condition, or (c) a control condition. The earlier significant outcome on a substance initiation index was replicated, and positive point-in-time results for weekly drunkenness were observed, but there were no statistically significant outcomes for regular alcohol use. Discussion focuses on factors relevant to the mix of significant longitudinal results within a consistent general pattern of positive intervention control differences. PMID- 16366810 TI - Risk and protective factors for substance use among African American high school dropouts. AB - Risk and protective factors that predict substance use were investigated with 318 African American high school dropout youths who completed the 1992 follow-up of the National Educational Longitudinal Study of 1988. A conceptual model linking positive family relationships and religious involvement to youths' substance use and conventional peer affiliations through a positive life orientation was examined with structural equation modeling. Positive life orientation, which included optimism and conventional goals for the future, fully mediated the influence of family relationships on conventional peer affiliations. Religious involvement directly predicted conventional peer affiliations and positive life orientation. Conventional peer affiliations mediated the other variables' influence on substance use. PMID- 16366811 TI - Moving beyond the keg party: a daily process study of college student drinking motivations. AB - Theoretical models of alcohol consumption assert that young adults endorse multiple drinking motives, including drinking to cope with negative experiences and to enhance positive experiences. Social contacts may be important to both pathways. This study applied daily process methodology to determine the relationship between college student drinking in different contexts and daily social contacts and moods. Each afternoon for 3 weeks, 122 undergraduates (43% men, 57% women) logged onto a secure Web site during specified hours to report daily activities, moods, and contacts. Hierarchical linear modeling analyses provided support for motivational models and the context-specific nature of motivated drinking. Individual differences were revealed for each motivation. These findings highlight the importance of studying within-person processes using daily process designs. PMID- 16366812 TI - Concordance among biological, interview, and self-report measures of drug use among African American and Hispanic adolescents referred for drug abuse treatment. AB - This study examined the concordance among urine assays, interview measures, and self-report measures of marijuana and cocaine use among 190 drug abusing/dependent African American and Hispanic adolescents and their families at 3 assessment points of an 18-month randomized clinical trial study. Results demonstrated concordance among urine assays, a calendar method self-report measure (Timeline Follow Back [TLFB]), and a noncalendar method self-report measure (Adolescent Drug Abuse Diagnosis Scale). Diagnostic criteria of marijuana and cocaine abuse/dependence from a clinical structured interview (Diagnostic Interview Schedule for Children [DISC]) also converged, albeit weakly, with self report measures. Adolescent and parent reports on DISC marijuana abuse/dependence diagnostic criteria were related; however, collateral findings for DISC cocaine abuse/dependence diagnostic criteria were equivocal. Differences in concordance among biological and self-report cocaine use measures were found for baseline TLFB assessments among African American participants. Implications for future use and refinement of adolescent drug use assessments are discussed. PMID- 16366813 TI - A follow-up psychometric analysis of the self-regulation questionnaire. AB - Self-regulation skills, which subsume goal-directed behavior and short-term delay of gratification for long-term gains, have been shown to be differentially related to alcohol consumption and alcohol-related consequences. J. M. Brown, W. R. Miller, and L. A. Lawendowski (1999) described the Self-Regulation Questionnaire (SRQ), and K. B. Carey, D. J. Neal, and S. E. Collins (2004) provided preliminary psychometric evidence for the SRQ and proposed a short version (SSRQ) of the measure. The goals of this study were to further examine the psychometric properties of the SSRQ. Participants (N = 237) were recruited from an introductory psychology course and completed a questionnaire packet that included the SSRQ. Factor analyses indicated that the SSRQ showed 2 distinct factors, an Impulse Control factor and a Goal Setting factor. Validity evidence showed differential patterns of relations between these 2 subscales and measures of self-control, alcohol use, and alcohol-related consequences. PMID- 16366815 TI - The effects of nicotine on attention and working memory in never-smokers. AB - The subjective and physiological effects of nicotine in nicotine-naive individuals are consistent across studies, though the cognitive effects are variable: Positive, negative, or no effects have been reported. Assessing specific cognitive processes (e.g., alerting, orienting, executive function, and phonological and visuospatial working memory) may help reduce this variability. This within-subject study (N = 20) was designed to assess the effect of nicotine gum (0, 2, or 4 mg) on subjective, physiological, and cognitive measures. Dose dependent increases in dysphoria and heart rate were observed, though nicotine did not influence any aspect of attention or working memory. Future studies should take into account the difference in effect sizes for cognitive versus physiological/subjective measures and maximize power (e.g., increase sample size) accordingly. PMID- 16366814 TI - Children's hedonic judgments of cigarette smoke odor: effects of parental smoking and maternal mood. AB - Age-appropriate tasks were used to assess 3- to 8-year-old children's liking, identification, and preference for a variety of odors, including that of exhaled cigarette smoke. Children whose parents smoke took longer to decide whether they liked the cigarette odor and were significantly more likely to prefer the odor of cigarette to the neutral and unfamiliar odor of green tea compared with children of nonsmokers. Among children of smokers, relative preferences for the cigarette odor were related to maternal mood disturbance and depression scores. These findings suggest that some early learning about cigarette smoke odor is based on sensory experiences at home and anchors it to the emotional context in which their mothers smoke. PMID- 16366816 TI - Ever users versus never users of a "less risky" cigarette. AB - The authors surveyed 173 smokers who purchased and 114 smokers who knew about but did not purchase Eclipse--a tobacco industry product that is advertised as a reduced risk cigarette. Most low-tar/nicotine cigarette users who purchased Eclipse believed Eclipse was safer than low-tar/nicotine cigarettes both for their own health (73%) and for the health of others around them (86%). Additionally, many viewed Eclipse as a step toward quitting (53%). The authors did not identify robust predictors of Eclipse use. At 6 months follow-up, Eclipse users were as likely to have tried to quit as nonusers; however, the small sample size does not rule out the possibility that Eclipse use undermines quit attempts. The authors conclude that almost all users believe Eclipse is safer than low-tar cigarettes. PMID- 16366817 TI - Sex differences in intrinsic aptitude for mathematics and science?: a critical review. AB - This article considers 3 claims that cognitive sex differences account for the differential representation of men and women in high-level careers in mathematics and science: (a) males are more focused on objects from the beginning of life and therefore are predisposed to better learning about mechanical systems; (b) males have a profile of spatial and numerical abilities producing greater aptitude for mathematics; and (c) males are more variable in their cognitive abilities and therefore predominate at the upper reaches of mathematical talent. Research on cognitive development in human infants, preschool children, and students at all levels fails to support these claims. Instead, it provides evidence that mathematical and scientific reasoning develop from a set of biologically based cognitive capacities that males and females share. These capacities lead men and women to develop equal talent for mathematics and science. PMID- 16366818 TI - Graduate study in psychology: 1971-2004. AB - This article traces the evolution of graduate study in psychology in the United States and Canada during the past 3 decades. The authors summarize the 2003-2004 characteristics of graduate programs and departments in psychology and compare these data to those obtained in 1970-1971, 1979-1980, and 1992-1993. The most recent data were obtained from the 2005 edition of Graduate Study in Psychology and were based on 495 institutions, 601 departments, and 1,970 graduate programs. Information is presented for both entire departments (e.g., faculty characteristics, student profiles, admission criteria, Graduate Record Examination and grade point averages, tuition costs, financial assistance) and individual graduate programs (e.g., areas of study, number of applicants, acceptance rates, enrollment statistics, retention rates). Particular attention is devoted to the emergence of PsyD programs in professional psychology, the ascendancy of neuroscience programs in academic psychology, and the steady rise in acceptance rates across programs over the past 33 years. PMID- 16366819 TI - Determination and documentation of the need for practice guidelines. PMID- 16366829 TI - Relationship between U.S. News and World Report's and the National Research Council's ratings/rankings of psychology departments. PMID- 16366830 TI - Evolutionary psychology and false confession. PMID- 16366831 TI - Some thoughts on the "staircase to terrorism". PMID- 16366832 TI - Historical conflict and incitement also provoke the journey to terrorism. PMID- 16366833 TI - Psychological processes and "the staircase to terrorism". PMID- 16366834 TI - More problems with gap closing philosophy and research. PMID- 16366836 TI - Highlights & happenings: response to Hurricane Katrina exposes emergency preparedness problems. PMID- 16366837 TI - Interview with Penny Bevan, MD, Head of the Emergency Preparedness Division, Department of Health, United Kingdom,(Interview by Madeline Drexler). PMID- 16366838 TI - National strategy for Pandemic Influenza and the HHS Pandemic Influenza Plan: thoughts and comments. PMID- 16366839 TI - Biological threat characterization research: a critical component of national biodefense. AB - Biological warfare (BW) threat assessments identify and prioritize BW threats to civilian and military populations. In an ideal world, they provide policymakers with clear and compelling guidance to prioritize biodefense research, development, testing, evaluation, and acquisition of countermeasures. Unfortunately, the biodefense community does not exist in an ideal world. National security professionals responsible for crafting BW threat assessments often are challenged by factors that limit the clarity and/or timeliness of those assessments. Moreover, the potential for life science advances to enhance threats enabled by state programs and the possibility that non-state actors may pursue crude but effective BW methodologies will drastically expand the scope of the perceived threat. Appropriate investment of federal biodefense funds will require some mechanism for validating and prioritizing present and future threats. Ideally, such a mechanism will incorporate empirical data targeted to elucidate actual hazards. In this regard, the Department of Homeland Security's creation of a Biological Threat Characterization Program for the technical validation of threat agents will be a valuable addition to the nation's overall biodefense strategy. This article articulates the need for a coordinated national biological threat characterization program, discusses some of the principal challenges associated with such research, and suggests a few options for their resolution. PMID- 16366840 TI - Leveraging bioterrorism preparedness for non-bioterrorism events: a public health example. AB - When a local health department in Virginia learned that a deceased hospital-based nurse had worked for several months with undiagnosed and untreated active tuberculosis, it mounted an extraordinary effort to find, screen, test, and potentially treat numerous contacts. In responding to this challenge, it adapted plans, concepts, and equipment that had been recently developed or acquired for responding to acts of bioterrorism. The improved coordination and integration with community partners and participating agencies, fostered through bioterrorism preparedness planning, were keys to success. Using procedures developed and exercised to distribute prophylactic medication in the Strategic National Stockpile, the health department shifted philosophically from a program-specific response to a more integrated approach. Implementation of this mass tuberculosis screening program was based on the principles of the Incident Command System (ICS). Owing to the efficiency of the operation, more than 2,500 people were quickly screened, and the rate of return for skin test readings was 91.6%, ranking it extremely high compared to the benchmarks. Overall, 5.9% of those tested were found to be infected with tuberculosis, and no cases of active tuberculosis were identified. This outcome demonstrated public health's improved ability to react, as a result of bioterrorism preparedness activities, to "traditional" public health mass events and non-bioterrorism emergencies. PMID- 16366841 TI - A psychosocial risk assessment and management framework to enhance response to CBRN terrorism threats and attacks. AB - Evidence in the disaster mental health literature indicates that psychosocial consequences of terrorism are a critical component of chemical, biological, radiological, and nuclear (CBRN) events, both at the clinical level and the normal behavioral and emotional levels. Planning for such psychosocial aspects should be an integral part of emergency preparedness. As Canada and other countries build the capacity to prevent, mitigate, and manage CBRN threats and events, it is important to recognize the range of social, psychological, emotional, spiritual, behavioral, and cognitive factors that may affect victims and their families, communities, children, the elderly, responders, decision makers, and others at all phases of terrorism, from threat to post-impact recovery. A structured process to assist CBRN emergency planners, decision makers, and responders in identifying psychosocial risks, vulnerable populations, resources, and interventions at various phases of a CBRN event to limit negative psychosocial impacts and promote resilience and adaptive responses is the essence of our psychosocial risk assessment and management (P-RAM) framework. This article presents the evidence base and conceptual underpinnings of the framework, the principles underlying its design, its key elements, and its use in the development of decision tools for responders, planners, decision makers, and the general public to better assess and manage psychosocial aspects of CBRN threats or attacks. PMID- 16366842 TI - The willingness of U.S. Emergency Medical Technicians to respond to terrorist incidents. AB - A nationally representative sample of basic and paramedic emergency medical service providers in the United States was surveyed to assess their willingness to respond to terrorist incidents. EMT's were appreciably (9-13%) less willing than able to respond to such potential terrorist-related incidents as smallpox outbreaks, chemical attacks, or radioactive dirty bombs (p<0.0001). EMTs who had received terrorism-related continuing medical education within the previous 2 years were twice as likely (OR=1.9, 95% CI 1.9, 2.0) to be willing to respond to a potential smallpox dissemination incident as those who indicated that they had not received such training. Timely and appropriate training, attention to interpersonal concerns, and instilling a sense of duty may increase first medical provider response rates. PMID- 16366843 TI - Updating the International Health Regulations. AB - First adopted in 1951, the International Health Regulations (IHR) provide the international legal framework for efforts to prevent and control the cross-border spread of communicable diseases. In 1995, after outbreaks of emerging infections had rendered the IHR increasingly obsolete, the 192 member states of the World Health Organization (WHO) requested a major updating of the regulations to adapt them to the highly mobile, globalized world of the 21st century. After negotiations in 2004 and 2005, the revised IHR text was adopted unanimously by the World Health Assembly, WHO's highest policymaking body. This article reviews the 2005 regulations and discusses their implications for the international response to natural epidemics and to incidents involving the accidental or deliberate release of biological or chemical agents or radiological materials. PMID- 16366844 TI - Evaluation of public health interventions for Anthrax: a report to the secretary's council on Public Health Preparedness. AB - To aid in understanding how best to respond to a bioterror anthrax attack, we analyze a system of differential equations that includes a disease progression model, a set of spatially distributed queues for distributing antibiotics, and vaccination (pre-event and/or post-event). We derive approximate expressions for the number of casualties as a function of key parameters and management levers, including the time at which the attack is detected, the number of days to distribute antibiotics, the adherence to prophylactic antibiotics, and the fraction of the population that is preimmunized. We compare a variety of public health intervention policies in the event of a hypothetical anthrax attack in a large metropolitan area. Modeling assumptions were decided by the Anthrax Modeling Working Group of the Secretary's Council on Public Health Preparedness. Our results highlight the primary importance of rapid antibiotic distribution and lead us to argue for ensuring post-attack surge capacity to rapidly produce enough anthrax vaccine for an additional 100 million people. PMID- 16366845 TI - Eight strategies to engage industry in biosecurity. AB - A significant portion of the domestic bio-research base-and the one most likely to provide translational research-is not engaged in biodefense. Despite the fact that more than one-third of all life science researchers are employed in commercial bio-research, fewer than 3% of the 1,500 U.S. bio-technology companies are involved in biosecurity initiatives. The bio-tech industry has largely not aligned itself to play an integral role in biosecurity, but there are a few policy changes that could dramatically alter this balance. These include engaging and motivating the bio-technology middle class, seeding secondary markets, focusing on system solutions, providing reagents and standards, aligning communications, and prioritizing translational research. By reaching out, policymakers can span the current chasm between the bio-industry and government, build a stable biodefense industrial base, establish solid working relationships, and secure better services and products. The rewards would be significant for government and industry alike. PMID- 16366846 TI - Bulls, Bears, and Birds: Preparing the Financial Industry for an Avian Influenza Pandemic. AB - Bulls, Bears, and Birds: Preparing the Financial Industry for an Avian Influenza Pandemic was a half day symposium on avian influenza for senior leaders and decision makers from the financial sector with responsibility for business continuity, health, and security. The event brought together experts and leaders from the medical, public health, business continuity, and financial communities to appraise financial industry leaders on the threat of avian influenza and to offer suggestions regarding what the financial industry could do to prepare and respond. PMID- 16366851 TI - Foodborne disease trends and reports. PMID- 16366852 TI - Evolution of foodborne pathogens via temperate bacteriophage-mediated gene transfer. AB - Temperate bacteriophages have always been central to the evolution of bacteria, although their importance has been consistently underestimated compared to transformation and conjugation. In the last 20 years, as more gene and genome sequences have become available and researchers have more accurately determined bacteriophage populations in the environment, we are gaining a clearer picture of their role in the past and potential role in the future. The transductive and lysogenic capacities of this class of bacteriophages have contributed to the evolution and shaping of emerging foodborne pathogenic bacteria through the dissemination of virulence and antibiotic resistance genes. For example, the genome sequences of Shigella dysenteriae, Escherichia coli O157:H7, and the Stxencoding bacteriophages demonstrate the critical role bacteriophage-mediated gene transfer events played in the evolution of these high-profile human pathogens. In this review, we describe the basic genetic exchange mechanisms mediated by temperate bacteriophages and how these mechanisms have been central to the dissemination of virulence genes, such as toxins and antibiotics from one species to another (the shiga-like toxins, and multiple antibiotic resistance dissemination in Salmonella are used as specific examples). Data demonstrating the role of bacteriophages in the spread of antimicrobial resistance in bacteria, including interspecies transduction, are also presented. That temperate bacteriophages play a role in the on-going evolution of emerging pathogenic bacteria is obvious, but it is also clearly an on-going process with a breadth that must be appreciated as well as studied further if we are to be able to foresee what new challenges will arise to imperil food safety. PMID- 16366853 TI - Susceptibility of Escherichia coli from growing piglets receiving antimicrobial feed additives. AB - Concerns regarding an apparent association between the use of antimicrobial feed additives (AFAs) in food animal production and a concomitant increase in antimicrobial drug resistance among zoonotic enteropathogens have provided the impetus to propose cessation of their use. While AFAs have been used in food animal production for nearly 50 years, the future use of AFAs will require an understanding of the effects of different classes of antimicrobials on the antimicrobial resistance of commensal flora. The present study examines the effect of three AFAs (apramycin, carbadox, and chlortetracycline) on the antimicrobial susceptibility of Escherichia coli in growing piglets and on animal performance. Three replicate trials were conducted using growing piglets fed standard diets with and without antimicrobial feed additives (AFAs). Fecal samples were cultured selectively for E. coli at regular intervals from all piglets from birth to market and antimicrobial susceptibility testing of E. coli isolates was performed using a replica-plate screening method and a broth microdilution method. While resistance to tetracycline in E. coli varied widely by sample, group, and trial, a significant increase in the percentage of resistant isolates was observed in piglets receiving AFAs when compared to controls (p < 0.0001). Resistance to apramycin increased in E. coli from piglets fed apramycin when compared to controls (p < 0.0001). However, upon removal of apramycin, resistance in E. coli declined to baseline levels by day 75. Piglets receiving AFAs demonstrated improved feed efficiency during phase 4 (p < 0.001), and higher average daily gains in phases 3 and 4 (p < 0.0001). This study suggests that antimicrobial resistance to AFAs in E. coli is drug-dependent and that some antimicrobials may be suitable for continued use in feeds during specified growth periods without concern for persistence of resistant E. coli populations. PMID- 16366854 TI - Effect of porcine-derived mucosal competitive exclusion culture on antimicrobial resistance in Escherichia coli from growing piglets. AB - While use of antimicrobial drugs in livestock production has made a significant impact on animal health, welfare, and productivity, interest in suitable alternatives such as pre/probiotics, organic acids, and cultures of normal flora or "competitive exclusion" cultures from young animals has increased significantly in the wake of the antimicrobial resistance issue. The present study was undertaken to determine the effect of porcine-derived mucosal competitive exclusion (PCE) culture on both the antimicrobial susceptibility of commensal E. coli and on growth performance in piglets. Two replicate trials were conducted using growing piglets fed standard antimicrobial-free production diets. Piglets in the treatment group were orally dosed with PCE (10(10) cfu/mL) twice within 24 h of birth, at weaning, and 18-24 h post-weaning; control group piglets were dosed with sterile broth as a placebo. Fecal samples from all piglets were cultured for commensal E. coli at dosing times and when feed type was changed. A significantly higher proportion of E. coli from PCE-treated piglets demonstrated resistance to tetracycline (p < 0.0001), and streptomycin (p < 0.0001) when compared to controls. Resistance to streptomycin resistance in E. coli from piglets treated with PCE culture was variable, returning to baseline levels by day 21 (weaning). Piglets treated with the PCE culture demonstrated improved feed efficiencies when compared to control piglets (p < 0.005) during feeding of the starter and first growth diets. The PCE culture used in the present study had previously been shown to effectively exclude Salmonella in pigs. To the best of the authors' knowledge, this is the first report characterizing the effect of a competitive exclusion culture on antimicrobial resistance of commensal E. coli. PMID- 16366855 TI - A comparison of the antimicrobial activity of garlic, ginger, carrot, and turmeric pastes against Escherichia coli O157:H7 in laboratory buffer and ground beef. AB - The antimicrobial effects of garlic, ginger, carrot and turmeric pastes against Escherichia coli O157:H7 in laboratory buffer and model food system were investigated. Turmeric paste, fresh carrot, ginger and garlic pastes from roots, and commercial ginger and garlic paste were heated alone or with buffered peptone water (BPW) or ground beef at 70 degrees C for 7 min. All samples were inoculated with a three strain cocktail of overnight cultures of E. coli O157: H7 and stored at 4 degrees C and 8 degrees C for 2 weeks. Each paste exhibited different antimicrobial effects alone and in ground beef or BPW at 4 degrees C and 8 degrees C for 2 weeks. Commercial ginger paste and fresh garlic paste showed the strongest antimicrobial activity with complete inactivation of E. coli O157:H7 in the paste at 3 days at 4 degrees C and 8 degrees C. Carrot and turmeric pastes did not show any antimicrobial activity both at 4 degrees C and 8 degrees C. Commercial garlic showed antimicrobial activity at both 4 degrees C and 8 degrees C (about 1 log CFU/g reduction) in the paste. However, fresh ginger paste showed antimicrobial activity only at 8 degrees C. Only commercial ginger paste had antimicrobial activity in BPW at 4 degrees C for 2 weeks. However, commercial ginger paste showed antimicrobial activity in ground beef at 3 days and after (about 1-2 log CFU/g) compared to control samples at 8 degrees C for 2 weeks. Fresh garlic paste showed antimicrobial activity only in BPW (1.3 log CFU/g) at 8 degrees C. These results indicate that the antimicrobial activity of these pastes is decreased in ground beef and laboratory buffer. PMID- 16366856 TI - Characterization of Escherichia coli and Salmonella enterica from cattle feed ingredients. AB - The objectives of this study were (1) to evaluate the frequency with which feed ingredients or mixed feeds in cattle feedlots were contaminated with Escherichia coli and Salmonella spp. and (2) to evaluate the antimicrobial susceptibility patterns of non-type-specific Escherichia coli and Salmonella spp. recovered from feed ingredients or mixed feeds. Approximately 30 individual samples were collected from each of several feed commodities present on two cattle feedlots each month for 1 year. Half of the samples were cultured for Escherichia coli, and the other half were cultured for Salmonella spp. E. coli was recovered from 48.2% (516/1070) of the samples and from all feed ingredient types at least once. Salmonella spp. were recovered from 5.3% (57/1070) of samples. Overall, 40.3% (207/514) of E. coli isolates and 54.4% (31/57) of Salmonella spp. isolates were susceptible to all antimicrobials tested in the panel. Bacterial contamination of feed ingredients used at cattle feedlots with enteric bacteria is relatively common. In some cases, the enteric organisms are resistant to one or more antimicrobials. Feed ingredients may be a source of genetic elements associated with antimicrobial resistance for feedlot cattle. To be successful in minimizing foodborne pathogens and antimicrobial resistance in the cattle feedlot setting, it is important to consider the myriad of potential sources of these organisms or genetic elements. PMID- 16366857 TI - Antimicrobial resistance and class 1 integrons in pathogenic Escherichia coli from dairy farms. AB - The goal of this study was to assess the prevalence of antimicrobial resistance and class 1 integrons, including integron-associated genes, in 24 Escherichia coli isolates from dairy farms. Escherichia coli isolates (n = 14) from dairy cows with mastitis (ECDM), Shiga toxin-producing (STEC) O157:H7 from cull dairy cow fecal samples (n = 9) and bulk tank milk (n = 1) were evaluated for sensitivity to 19 antimicrobial agents used commonly in human and/or veterinary medicine. Multiplex PCR was used to determine presence of genes associated with class 1 integrons (intI1, qacEDelta1, and sulI1). Class 1 integrons were found only in eight of 10 isolates (one STEC O157:H7 and seven ECDM) that demonstrated antimicrobial resistance, and seven of these were resistant to two or more antimicrobial agents. Eight of 10 STEC O157:H7 and six of 14 ECDM were susceptible to all commonly used antibiotics. Five ECDM demonstrated multiple resistances to four or more antibiotics. Most of the 24 isolates examined exhibited resistance against sulfamethoxazole, followed by streptomycin and tetracycline. STEC O157:H7 strains had less prevalence of antibiotic resistance and integron carriage than ECDM. The multiplex PCR method developed for detection of intI1, qacEDelta1, and sulI1 can be used routinely for monitoring presence of these genes. Class 1 integrons were found in eight of 10 E. coli strains that demonstrated antimicrobial resistance; seven of these were resistant to two or more antibiotics. It appears that integrons played a role in the incidence of antimicrobial resistance of the strains used in this study. PMID- 16366858 TI - Survival, injury, and virulence of freeze-stressed plasmid-bearing virulent Yersinia enterocolitica in ground pork. AB - The effect of freezing at -20 degrees C on survival, injury, stability of the virulence plasmid, and expression of virulence-associated determinants of Yersinia enterocolitica (YEP(+)) in ground pork was assessed. The viable counts on nonselective and selective media after 12 weeks of freezing were similar to counts obtained before freezing, indicating that the YEP(+) strain survived the freezing process. There was no evidence of freeze-stress injury. Results of a PCR assay targeting plasmid-associated virF gene and virulence determinants indicate that YEP(+) subjected to freezing are potentially capable of causing food-borne illness and freezing is not a substitute for safe handling and proper cooking of pork. PMID- 16366859 TI - Prevalence and characterization of Escherichia coli O26 and O111 in retail minced beef in Ireland. AB - Verocytotoxigenic Escherichia coli (VTEC) O157 are recognized as bacterial pathogens with significant public health impact. However, other serogroups, including O26, O111, O103, and O145, have the potential to cause the same spectrum of illness. In this study, 800 minced (ground) beef samples covering a large geographical region in Ireland were collected and tested for Escherichia coli (E. coli) O26 and E. coli O111 by conventional microbiological protocols. Two minced beef samples (0.25%) tested positive for E. coli O26, indicating fecal contamination. None of these isolates possessed verocytotoxin-encoding genes, (vt1/vt2 also known as stx1/stx2), the hemolysinencoding gene (hlyA), or the E. coli attachment-effacement (eae) gene, as determined by polymerase chain reaction (PCR). None of the beef samples analyzed contained E. coli O111. Although the E. coli O26 isolates were nonvirulent, the presence of these isolates in raw minced beef is an indication of fecal contamination and therefore potentially of public health significance. PMID- 16366860 TI - Resistance to oxyiminocephalosporins mediated by blaTEM-52 genes in Salmonella typhimurium from humans in England and Wales. AB - The emergence of antimicrobial resistance among Salmonella is a matter of great public health concern, more so in the case of extended-spectrum cephalosporins, since these antimicrobials are normally regarded as the drugs of choice for complicated cases of infection. This study was designed to determine the occurrence of resistance mediated by the presence of extended-spectrum beta lactamases (ESBL) enzymes belonging to the TEM family. Only two isolates were detected after analysis of the 278,308 Salmonella isolates from the last 10 years. In both cases, the gene involved was a bla (TEM-52)-like, and infections were linked with foreign travel. ESBL-TEM enzymes remain very rare in Salmonella in England and Wales, and no domestic cases have been detected to date. PMID- 16366861 TI - Eye motion parameters correlate with level of experience in video-assisted surgery: objective testing of three tasks. AB - BACKGROUND: Laparoscopic skills vary with experience and training; however, objective measures to ascertain the level of training have not yet been established. New technology allows noninterfering measurement of eye motion parameters that correlate with attention and distraction during visually oriented tasks. Our objective was to apply this new technology in the setting of video assisted surgery to evaluate eye motion parameters among surgeons of varying experience. MATERIALS AND METHODS: Subjects with various levels of laparoscopic experience (novice, intermediate, and expert) were fitted with a noninvasive, Food and Drug Administration approved, eye motion monitoring device. The device was used to measure and record parameters of eye motion, including saccadic rate (SR), standardized peak velocity (PV), standardized saccadic amplitude (SA), and the duration of gaze fixation (FD), during the performance of 3 basic laparoscopic tasks on a laparoscopic training station. RESULTS: A total of 24 subjects (3 groups of 8 each) participated in this study. Experience level was found to have a main significant effect on SR (P = 0.047) and PV (P = 0.028). Two way ANOVA demonstrated that experience level approached significance for SA (P = 0.058) and FD (P = 0.055). CONCLUSION: The advancement of laparoscopic techniques and instrumentation relies, in part, on expanding the current understanding of operator/instrument interactions. This places an increasing demand on objective methods of monitoring such interactions during laparoscopy. Our study demonstrates a significant difference in eye motion parameters in surgeons with differing levels of experience. Further testing is needed in actual clinical settings to determine the importance of eye motion during surgery. PMID- 16366862 TI - Postoperative adhesion formation after laparoscopic uterine horn resection in a porcine model: comparison of five instruments. AB - PURPOSE: To evaluate the postoperative adhesion formation caused by instruments used in gynecologic laparoscopic surgery and to determine the optimal instruments to use to reduce adhesions. MATERIALS AND METHODS: Seventeen juvenile pigs underwent laparoscopic bilateral resection of the uterine horns under general anesthesia and pneumoperitoneum. The laparoscopic procedures were carried out using monopolar electrocautery (ME) (n = 8), an electrothermal bipolar vessel sealer (EBVS) (n = 6), an ultrasonically activated scalpel (UAS) (n = 6), a loop type ligature (LTL) with a steel scalpel for severing the tissues (n = 6), and an automatic stapling device (ASD) (n = 8). Second- look laparotomy was performed 14 days postoperatively, and the degree of postoperative adhesions was scored from 0 to 6. RESULTS: The mean and range of adhesion scores were 0.00 with EBVS, 0.13 (range, 0-1) with ASD, 0.33 (range, 0-2) with LTL, 1.17 (range, 0-3) with UAS, and 3.13 (range, 2-6) with ME. We found a statistically significant difference in the extent of postoperative adhesion formation associated with these 5 instruments (P < 0.001, Kruskal-Wallis test). CONCLUSION: Adhesion formation increased in the order EBVS < ASD < LTL < UAS < ME. Our study strongly suggests that surgical instruments can be selected to reduce postoperative adhesion formation, a particular concern in women of reproductive age. PMID- 16366863 TI - Endoscopic extraperitoneal inguinal hernia repair with double mesh: indications, technique, complications, and results. AB - Although totally extraperitoneal laparoscopic hernia repair has the same benefits attributed to the traditional preperitoneal prosthetic surgical repair, this procedure is not used widely because of perceived difficulty in dissection. Since one of the most common causes of hernia recurrence in this procedure is inadequate lateral inferior and medial inferior mesh fixation, we have introduced a double-mesh technique in an effort to reduce the rate of recurrence. Our procedure is a variation of the totally extraperitoneal laparoscopic inguinal hernia repair and provides a more secure inguinal floor by adjusting the second mesh to the area of weakness. We describe the laparoscopic inguinal hernia repair by the extraperitoneal double-mesh technique performed in 53 selected patients with very large indirect hernias and extremely large bilateral or recurrent hernias. The mean operative time was 74 minutes for unilateral hernias and 110 minutes for bilateral hernias. The median follow-up time was 65 months (range, 9 97 months) with no recurrences, neuralgia, or bleeding complications. We believe that this technique offers perfect positioning of the meshes and provides the most secure inguinal floor. Therefore, the method is presented for consideration in the laparoscopic repair of large indirect, direct, or recurrent hernias. PMID- 16366864 TI - Laparoscopic transperitoneal adrenalectomy using the LigaSure vessel sealing system. AB - BACKGROUND: Laparoscopic adrenalectomy is being performed with increasing frequency in the surgical treatment of adrenal tumors. Among laparoscopic approaches to the adrenal glands, the transperitoneal access is preferred. Along with advances in technology, different energy systems have been utilized. Laparoscopic adrenalectomy has become easier with the use of the LigaSure vessel sealing system (Valleylab, Boulder, Colorado). MATERIALS AND METHODS: Between January 2002 and August 2004, 23 laparoscopic transperitoneal adrenalectomies were performed in 22 patients using the LigaSure vessel sealing system: 16 of the patients were female, 6 were male and the mean age was 44 years (range, 17-70 years). Indications for surgery were non-functioning adenoma (n = 10), pheochromocytoma (n = 4), Cushing's syndrome (n = 5), Conn's syndrome (n = 2), and lymphoma (n = 1). The mean diameter of lesions was 4 cm (range, 1-7 cm). The distribution was 12 left, 9 right, and 1 bilateral adrenalectomies. RESULTS: The mean operative time for unilateral adrenalectomies was 57 minutes (range, 30-75 minutes). The operative time for the patient with Cushing's disease in whom a bilateral adrenalectomy was performed was 180 minutes. All operations were completed laparoscopically. A nonsteroidal anti-inflammatory drug (Lornoxicam) was sufficient for postoperative analgesia. Oral feeding was started 6 hours postoperatively. When used, drains were removed on postoperative day 1. The mean postoperative hospital stay was 1.5 days (range, 1-3 days). Wound infections developed in two patients with Cushing's syndrome. There was no mortality. Histopathologic examination of specimens revealed a cortex adenoma in 16 cases (10 of which was nonfunctional), a pheochromocytoma in 4 cases, a bilateral cortical hyperplasia in 1 case, and a lymphoma in 1 case. CONCLUSION: Laparoscopic adrenalectomy is an established method in the treatment of adrenal masses. Laparoscopic adrenalectomy as well as other laparoscopic procedures has become easier with the introduction of new energy systems. Vascular control and dissection of the gland by Liga- Sure is feasible. It makes the procedure easier and eventually shortens the operation time. PMID- 16366865 TI - A comparison of anterior and posterior approaches for the surgical treatment of pancreatic pseudocyst using laparoscopic cystogastrostomy. AB - BACKGROUND: Laparoscopic treatment of pancreatic pseudocyst allows for definitive drainage with faster recovery. Although many groups have reported their experience with an anterior approach, only a few have done so with a posterior approach. This paper compares the approaches, analyzing their potential benefits and pitfalls. MATERIALS AND METHODS: Seven females and one male underwent laparoscopic cystgastrostomy to treat pancreatic pseudocysts. The anterior approach was performed by opening the stomach anteriorly, localizing the pseudocyst ultrasonographically, draining the cyst with a needle and, via the same opening, using a stapler to form a cystgastrostomy. The posterior approach was performed by directly visualizing the posterior gastric wall and the pseudocyst, opening and draining the cyst with a needle, and using a stapler and running sutures for closure. RESULTS: All patients had gallstone pancreatitis. Cystgastrostomy via the anterior approach was used in 4 patients and via the posterior approach in 4 patients. Dense adhesions required one attempted posterior cystgastrostomy to be converted to an anterior approach. The mean age of the anterior group was 38 years (range, 18-58 years) and hospital stay was 6 days (range, 4-8 days): for the posterior group, mean age was 42 years (range, 40 44 years) and length of stay was 3 days (range, 2-4 days). CONCLUSION: Although both approaches had good results with no complications and short hospital stays, the posterior approach is safer, with a more precise cyst visualization and dissection that permits more tissue to be sent for histopathologic examination. Furthermore, the posterior approach?s larger anastomosis would seem to yield fewer occlusions, which are commonly seen with the anterior approach. The anterior approach is easier to learn, but it requires the opening of the anterior stomach and the use of ultrasound. PMID- 16366866 TI - Laparoscopic repair of ventral and incisional hernias: our experience in 150 patients. AB - BACKGROUND: Incisional hernias develop in 2 to 20% of laparotomy incisions. Approximately 100,000 ventral hernias are operated on each year in the United States. The reported recurrence rates for open repair are 25-52% for primary and 12.5-19% for mesh repair. Compared to the open technique, laparoscopic repair has low rates of complications and recurrence, greater patient acceptance, and shorter hospital stay. MATERIALS AND METHODS: Between April 1999 and April 2004, we treated 150 patients (111 female, 39 male) with ventral and incisional hernias laparoscopically. Data concerning the age and sex of patients, the location, number, and size of fascial defects, the type of hernias and their contents, the size and type of meshes used in repair, operative time, length of hospital stay, and postoperative complications were collected. RESULTS: Of the 150 patients, 92 had incisional hernias and 58 had umbilical hernias. Polypropylene was used in 85 cases, dual mesh in 40 cases, and composite mesh in 25 cases. The mean age of the patients was 56.0 years (range, 33-81 years). Mean operative time was 63 minutes (range, 30-125 minutes). Mean postoperative hospital stay was 2.5 days (range, 1 15 days). The postoperative complication rate was 8.6% (seroma, paralytic ileus, small bowel injury, and suture-site neuralgia). The mean follow-up period was 32 months (range, 4-60 months). There was a 3% recurrence rate. Three subileus cases were detected during follow-up. CONCLUSION: The laparoscopic approach to ventral and incisional hernias is safe, feasible, and a good alternative to the open approach. Our results are comparable with those of other reports in the literature. PMID- 16366867 TI - Bilateral laparoscopic radical nephrectomy for renal tumors in patients with acquired cystic kidney disease. AB - PURPOSE: We describe our experience with simultaneous bilateral laparoscopic radical nephrectomy performed in patients with acquired cystic kidney disease (ACKD) and renal tumors. MATERIALS AND METHODS: Between June 2000 and September 2002, 10 patients with ACKD underwent simultaneous bilateral laparoscopic radical nephrectomy for renal lesions suspicious for carcinoma. The lesions were discovered during pretransplant evaluation in 9 patients and incidentally in 1 renal transplant recipient. A 3- or 4-port transperitoneal approach was used for each side to mobilize the kidney and secure the renal hilum. Both specimens were extracted through a midline supraumbilical incision. Operative time, blood loss, analgesic requirements, hospital stay, and convalescence and recurrence rates were determined. RESULTS: The mean age of the patients was 41.6 years (range, 29 47 years). Mean operative time was 6.5 hours (range, 4.5-9.7 hours) and mean estimated blood loss was 164 cc (range, 50-300 cc). There was one intraoperative complication-a clotted arteriovenous (AV) graft; and 2 postoperative complications-1 fluid overload and 1 adrenal insufficiency. The average length of hospital stay was 3.1 days (range, 2-4 days) and mean convalescence was 2.8 weeks (range, 1-6 weeks). All cancers were confined to the kidneys and there has been no recurrence during follow-up ranging from 6 to 26 months. CONCLUSION: Bilateral laparoscopic radical nephrectomy in end-stage renal disease patients is safe and feasible. The advantages of the laparoscopic approach include minimal intraoperative blood loss, shorter hospital stay, minimal postoperative pain, and a rapid return to normal activity. The laparoscopic technique offers an effective, minimally invasive therapeutic alternative to open surgery in high risk end-stage renal disease patients. PMID- 16366868 TI - Hand-assisted laparoscopic low anterior resection for rectal carcinoma. AB - Laparoscopic low anterior resection for rectal carcinoma has never been widely accepted among general surgeons because of the technical difficulties encountered during pelvic dissection. We describe our technique of hand-assisted laparoscopic low anterior resection (HAL-LAR) for rectal carcinoma using the Lapdisc abdominal wall sealing device (Hakko Medical, Tokyo, Japan, and Ethicon Endo- Surgery, New Brunswick, New Jersey) which results in pelvic dissection almost equivalent to the laparotomic operation. Thirteen patients with rectal adenocarcinoma (lower edge less than 15 cm from the anal verge) underwent laparoscopic low anterior resection, including 8 standard laparoscopic low anterior resections (SL-LAR) and 5 HAL-LAR. The mean operative time in the HAL-LAR group (211 +/- 48 min) was significantly shorter than in the SL-LAR group (311 +/- 78 min) (P = 0.0268). The mean intraoperative blood loss in the HAL-LAR group (37 +/- 45 g) was less than that in the SL-LAR group (198 +/- 177 g) (P = 0.075). The mean distal margin in the HAL-LAR group (23 +/- 4.5 mm) was longer than in the SL-LAR group (15 +/- 13.1 mm) (P = 0.2199). One patient in the SL-LAR group was found to have anastomotic recurrence in the staple suture line 10 months after surgery and died from cancer 24 months after surgery. One patient in the SL-LAR group was converted to open surgery because the distal margin was insufficient. In the HAL LAR group, there were no intra- or postoperative complications, no conversion to open surgery, and no recurrence after surgery. PMID- 16366869 TI - Laparoendoscopic techniques for occult gastrointestinal bleeding. AB - Up to 5% of gastrointestinal bleeds occur between the ligament of Treitz and the ileocecal valve. These patients present with occult bleeding and pose diagnostic and therapeutic challenges. Currently, an array of technology exists for diagnostic purposes, including upper and lower endoscopies, capsule endoscopy, nuclear scans, angiography, and intraoperative endoscopy. All of these modalities have advantages and disadvantages. However, the diagnostic gold standard for occult gastrointestinal bleeding does not exist. We present a case of an 18-year old male with occult gastrointestinal bleeding to illustrate the variety of available modalities. Initially, the patient underwent upper, lower, and push endoscopies. Subsequently, he had a nuclear bleeding scan, video capsule endoscopy, and an angiogram. Ultimately, the patient had a diagnostic laparoscopy with mobilization of the terminal ileum and right colon. The bowel was exteriorized and an intraoperative endoscopy was performed through a small bowel enterotomy. Multiple arteriovenous malformations (AVMs) were ascertained and resected. This case is presented in detail and the technique of intraoperative endscopy is reviewed. The diagnostic literature regarding AVMs is also reviewed. PMID- 16366870 TI - Transcervical video-assisted resection of Zenker's diverticulum. AB - We report a novel technique for resection of symptomatic Zenker's diverticulum. Two consecutive patients underwent a transcervical video-assisted dissection and resection of Zenker's diverticular pouch. Both patients exhibited good anatomical and functional results at 10 months follow-up. PMID- 16366871 TI - Laparoscopic resection of a giant liver hemangioma. AB - We present a case report of hepatic resection performed laparoscopically for a giant symptomatic hemangioma. The patient was a 50-year-old woman who was diagnosed with a giant hemangioma 20 cm in diameter. As in the case of our patient, laparoscopic resection of hepatic hemangiomas can be performed successfully, even when dealing with giant hemangiomas. However, a thorough preoperative assessment and a structured approach, in our case including preoperative embolization, is crucial for success. PMID- 16366872 TI - Laparoscopic management of gallstone presenting as obstructive gangrenous appendicitis. AB - We present an unusual case of a 55-year-old man with symptoms of recurrent appendicitis. Laparoscopy revealed a 1.5 cm gallstone impacted at the base of the appendix, leading to gangrenous appendicitis. This patient did not have any features of gallstone ileus. On imaging he had an inflammatory mass in the region of the right iliac fossa with a hyperintense shadow in the cecal area which was reported as an appendicolith. There was no demonstrable cholelithiasis or biliary enteric fistula. There were dense omental adhesions in the pericholecystic area on laparoscopy. The case was successfully managed by laparoscopic appendectomy with retrieval of the gallstone. No surgery was undertaken for the gallbladder. Diagnosis was confirmed by biochemical analysis of the stone, which contained calcium bilirubinate and cholesterol. A gallstone obstructing the appendicular lumen is a very rare etiology of gangrenous perforation of the appendix peritonitis. This case was successfully managed laparoscopically. PMID- 16366873 TI - Endoscopic ultrasound diagnosis and laparoscopic excision of an omental lymphangioma. AB - Omental lymphangiomas are rare intra-abdominal tumors. We describe a case of an omental lymphangioma which appeared on computed tomography scan as a possible gastric duplication cyst. The lesion was correctly diagnosed preoperatively via endoscopic ultrasound and complete excision of the cyst was performed laparoscopically. PMID- 16366874 TI - Conservative treatment of a huge abdominal wall hematoma after laparoscopic appendectomy. AB - We report a case of a 30-year-old male patient who presented with a huge abdominal wall hematoma 60 minutes after laparoscopic appendectomy. During surgery there had been no sign of abdominal wall bleeding. We found a lateral abdominal wall hematoma caused by rupture of the abdominal epigastric artery after trocar insertion. We conclude that trocar injury is a potentially preventable complication in laparoscopic surgery if certain precautions are taken. Abdominal wall hematoma secondary to injury of an epigastric vessel can be successfully treated with conservative management. PMID- 16366875 TI - Laparoscopic bilateral hernia repair using fibrin sealant: technical report of two cases. AB - Minimally invasive surgery is widely used in hernia repair given its advantages such as minimal disturbance to the surrounding tissues, shorter hospital stay, and promising long-term results. Efforts are still being made to make this minimally invasive procedure even more minimal. New tissue adhesives avoid the use of foreign materials and the postoperative pain that might be attributed to staples. We present the first two cases of bilateral inguinal hernia repair performed with a totally extraperitoneal procedure using fibrin sealant instead of staples for the fixation of the mesh. PMID- 16366876 TI - Presidential Address from the International Pediatric Endosurgery Group (IPEG) 14th Annual Congress for Endosurgery in Children, Venice Lido, Italy, June 1-4, 2005. PMID- 16366877 TI - Novel uses of surgical robotics in head and neck surgery. AB - PURPOSE: To demonstrate the utility of robotically assisted approaches in head and neck surgery. MATERIALS AND METHODS: Two teenage patients, one with a solitary thyroid nodule who was scheduled for a right thyroid lobectomy and the other with intractable seizures who was scheduled for placement of a vagal nerve stimulator were offered the option of a robotically assisted technique using a transaxillary endoscopic approach. RESULTS: Both procedures were completed successfully using the da Vinci surgical system (Intuitive Surgical, Sunnyvale, California). A 12 mm telescope and 5 mm instruments were used. There was sufficient mobility of the robotic arms despite the small working space. There were no complications, minimal pain in the axillary incisions, and patient satisfaction was high. Operative times were 4.5 and 4.2 hours, respectively. CONCLUSION: Transaxillary, endoscopic, robotically assisted approaches to the head and neck are feasible. The addition of robotics improves surgical dexterity in a difficult-to-reach anatomic region. Patient satisfaction appears high because of the avoidance of a cervical incision. PMID- 16366878 TI - Laparoscopic evaluation of the contralateral groin in pediatric inguinal hernia patients: a comparison of 70- and 120-degree endoscopes. AB - BACKGROUND: Transinguinal laparoscopic groin evaluation using a 70-degree endoscope can obviate the need for a second incision when attempting to identify a contralateral patent processus vaginalis (PPV) during open repair of a symptomatic pediatric inguinal hernia. This technique can be technically unsatisfactory when a medial veil of peritoneum obscures adequate visualization of the internal inguinal ring. This study compared 70- degree and 120-degree endoscopes in identification of a contralateral PPV in the same patients. MATERIALS AND METHODS: From September 2000 to October 2003, 81 patients with known inguinal hernias underwent open hernia repair and transinguinal laparoscopic evaluation of the contralateral side. The patients were 62 male, 19 female; mean age 26 months (range, 1 month-10 years); mean weight 11.7 kg (range, 2-33 kg). There were 53 right side hernias and 28 left side. Mean operative time was 43 minutes. Fifty seven patients (70%) had one or more risk factors for developing a contralateral inguinal hernia (49 were younger than 1 year old and 19 were ex-premature). Nineteen patients underwent concurrent procedures (7 circumcisions, 10 hydrocelectomies, 1 orchidopexy, 1 appendectomy). RESULTS: Using the 70-degree endoscope, a medial veil of peritoneum made visualization of the internal inguinal ring impossible in 14 patients (17%) and difficult in an additional 5 patients (6%). Visualization with the 120-degree endoscope was deemed to be superior in 46 (57%), equal in 35 patients (43%), and inferior in none. Overall, contralateral PPVs were detected in 31 patients (38%) with the 120 degree endoscope and in only 23 patients (28%) with the 70-degree endoscope. Had we used only the 70-degree endoscope, 8 PPVs (10%) would have been missed. None of the negative 120-degree endoscope evaluations have developed symptomatic contralateral inguinal hernias. CONCLUSION: In this trial, transinguinal laparoscopic evaluation using the 120-degree endoscope provided superior visualization and identification of contralateral PPVs. This new technique obviates the need for a separate abdominal wall puncture, reduces missed contralateral PPVs, and should be considered for use during pediatric inguinal hernia repair. PMID- 16366880 TI - Primary thoracoscopic repair of diaphragmatic hernia with pericostal sutures. AB - The surgical management of congenital diaphragmatic hernias has traditionally been via laparotomy or thoracotomy. Although laparoscopic and thoracoscopic repairs have been described, most reports are in older infants. We describe a method for primary thoracoscopic repair applied in the immediate neonatal period when no posterolateral rim of diaphragm exists. This simple technique for placing the pericostal sutures is a useful adjunct in the thoracoscopic management of diaphragmatic hernias. PMID- 16366879 TI - Video-assisted thoracoscopy for diaphragmatic plication: experimental study in a canine model. AB - BACKGROUND: Plication of a nonfunctional hemidiaphragm usually restores altered ventilatory mechanics. This study compared two techniques in performing diaphragmatic plication: video-assisted thoracoscopy (group A) and thoracotomy (group B). MATERIALS AND METHODS: Twenty dogs with induced paralysis of the right hemidiaphragm were randomly assigned to one of the two groups. Evaluations were performed before and after plication of the respiratory frequency (f) and lung area (LA) of the affected side. Operative time, time to resumption of walking, ingestion of fluids and solid food, pain intensity, and postoperative complications were measured. RESULTS: Group A had less pain after the surgery (P < 0.0001), earlier fluid ingestion (P < 0.05), and earlier resumption of walking (P < 0.019). Four weeks after the procedure, LA was similar in both groups, while a significant decrease in f was recorded in group A (P < 0.02). The remaining evaluated variables showed no differences. CONCLUSION: Both approaches were effective. Pain recorded in the postoperative period was less and recovery was faster in group A. Complications and surgical times were similar. The video assisted thoracoscopy is a safe and efficient option for performing diaphragmatic plication in dogs. PMID- 16366881 TI - Laparoscopic splenopexy of a pediatric wandering spleen by creation of a retroperitoneal pocket. AB - Wandering spleen is a rare condition in which the spleen is attached by a long, vascular pedicle and lacks its usual peritoneal attachments and supporting ligaments. This condition predisposes the spleen to torsion and infarction. We report the case of a 2-year-old boy with a history of intermittent abdominal pain and early satiety who presented with abdominal pain and severe gastric distention. A work-up including computed tomographic and ultrasonographic imaging suggested a wandering spleen. The diagnosis was confirmed during laparoscopy, and splenopexy was performed by anchoring the spleen in a retroperitoneal pocket in the left upper quadrant at the level of the tenth rib. To our knowledge, this is the first reported case of minimally invasive splenopexy for wandering spleen that involves the creation of a retroperitoneal pocket without the use of mesh. The authors believe that this is a safe and effective method that takes advantage of laparoscopy and avoids the risk of infection and complications associated with the use of synthetic material. PMID- 16366882 TI - Benefits of the thoracoscopic approach for short- or long-gap esophageal atresia. AB - OBJECTIVE: To evaluate the feasibility and results of thoracoscopy in various types of esophageal atresia (EA). MATERIALS AND METHODS: From April 2001 to August 2002, 5 patients with EA were treated by thoracoscopy. Their mean gestational age was 38 weeks and mean birth weight was 2700 g. Two patients had short-gap atresia with tracheo-esophageal fistula (type III according to Ladd's classification). Three had long-gap atresia: 2 with low fistula to the carina (type IV) and 1 without fistula (type I). Patients were placed in a prone position with the right side elevated at 80 degrees . Four intrapleural ports were necessary. The fistula when present was dissected and sutured with intrathoracic knots and esophageal anastomosis performed in the same manner. RESULTS: Positive airway pressure increased in all patients after insufflation but was kept in a safe range to prevent lung injury. An esophageal anastomosis was performed in 3 cases (2 short gaps and 1 long gap). Oral feeding started on day 6, and their mean length of hospital stay was 14 days. For one child with type IV EA, the anastomosis was impossible because of a long gap confirmed by an immediate thoracotomy. The ends were just approximated. A "spontaneous" fistula developed, and normal feeding was possible 2.5 months later. For the child with type I EA, the pouches could be only approximated at 2 months of age. A spontaneous fistula developed with a stenosis. A redo anastomosis by open surgery allowed for normal feeding. CONCLUSION: The thoracoscopic repair of an esophageal atresia is a reasonable choice for experienced surgeons treating patients, including those with long gaps. PMID- 16366884 TI - Assessment of personality and demographic aspects of cohabitation and marital success. AB - The outcomes of 68 marriages of four-year duration were compared on the basis of whether the partners had or had not cohabited premaritally. Background characteristics and personality data were assessed on those couples when there were newly married, and they were followed-up four years later to determine their current marital status, level of satisfaction, difficulty with various problem areas and the number of children born to them. No reliable differences on marital satisfaction or divorce rates between premarital cohabitors and noncohabitors were found. Cohabitors who divorced did so while reporting less marital distress than noncohabitors who divorced. Premarital cohabitors had significantly fewer children than noncohabitors. Degree of difficulty experienced on various problem areas differed between the groups. Certain personality and background variables predicted marital success differently for the two groups. Using the same selected set of six predictor variables, multiple regression equations were generated for cohabitors and noncohabitors. The cohabitors equation (R2 = .56) predicted marital success significantly more effectively than the noncohabitors equation (R2 = .28), When comparing the tri-weight vectors for these two equations, none of the predictor variables had the same influence in both groups. Theoretical implications of the findings were discussed. PMID- 16366885 TI - Effects of "favorability" and type of assessment device upon acceptance of general personality interpretations. AB - College students' acceptance of general personality interpretations was evaluated as a function of type of test and assessment device employed and interpretation favorability. Unlike previous studies, only interpretation favorability influenced acceptance. The effect of interpretation favorability upon acceptance, however, was shown to be due to the higher base-rate accuracy of favorable, relative to unfavorable, interpretations. It was concluded that the universal validity of general interpretations may supersede the effects of situational factors upon acceptance. PMID- 16366886 TI - Fakability of an empirically derived selection instrument. AB - Fakability of an empirically derived selection instrument developed by criterion keying for a specific occupation and organization was investigated. Ninety-four subjects completed the test under both Honest and Fake instructions. Instructions to simulate a highly motivated job applicant produced significant score increases on 7 of 10 scales and significant changes in narrative statements of predicted job performance based on these scores. Theoretical and practical implications are discussed. The caveat to use tests only when the subject is motivated to answer honestly is repeated. PMID- 16366896 TI - Toward a construct validation of the Rorschach. AB - The debate over the validity of the Rorschach has been lengthy but unresolved. Major reasons for the lack of consensus and conclusion are the absence of agreement about, how validation research should be conducted and the failure to identify appropriate criteria for subsequent evaluation. A construct validation model for the Rorschach is presented, with guidelines for proper implementation. The difficulties in establishing the optimal relationship between theory, research design, and clinical practice arc discussed. Recommendations for a resolution of these difficulties are presented. PMID- 16366897 TI - Canonical versus factor analytic perspectives on the structure of associations between the MMPI and the Buss-Durkee Hostility Inventory. AB - A canonical correlational analysis of MMPI profiles versus Buss-Durkee hostility and aggression scales was conducted, resulting in the identification of a general hostility-aggression-maladjustment dimension and three relatively specific aggression-personality relationships. These findings were contrasted with those obtained from a factor analysis of the combination of both sets of scores. Though often used in previous studies of associations between the Buss-Durkee Hostility Inventory and other measures, factor analysis was seen to be less than optimally suited to such a purpose. It was concluded that canonical correlation is preferable to factor analysis when seeking to clarify the multivariate structure of relationships between two sets of variables. PMID- 16366898 TI - Measurement of supernatural belief: sex differences and locus of control. AB - Although we live in an age dominated by science and technology, there exists an increasingly popular anti-science sentiment. This study describes the development of a scale to assess the degree of personal acceptance of supernatural causality versus acceptance of scientific explanation. In addition to the psychometric data concerning validity and reliability of the scale, data are presented which showed the personality factor of supernaturalism to be independent of orthodox religious attitudes. Results indicated a significantly greater supernatural acceptance for women, and a positive relation of supernaturalism with external locus of control. PMID- 16366899 TI - On the two-dimensional nature of the Marlowe-Crowne Social Desirability Scale. AB - Ramanaiah, Schill, and Leung (1977) reported empirical evidence supporting Millham's (1974) hypothesis that attribution and denial are two separate components of the MC-SD scale. This paper presents the results of two studies conducted to test the hypothesis that the results obtained by Ramanaiah et al. (1977) may be attributed to method variance caused by the keying direction in the MC-SD attribution and denial subscales. The first study was concerned with the development of balanced attribution and denial scales, while the second study investigated whether the results from Ramanaiah et al., which used the original unbalanced attribution and denial scales, could be replicated with the balanced scales developed in Study I. The results strongly supported the tested hypothesis, indicating that the two subscales are measuring essentially the same construct. PMID- 16366900 TI - A microgenetic approach to the integration of contrasexual characteristics. AB - Assessed relationships between introversion-extraversion and the integration of contrasexuality. Pictorial stimuli containing contrasexual or congruent sexual content were presented tachistoscopically at increasing durations, with recognition time scores and the rated emotional distance levels of percept descriptions selected as indices of integration. The following hypotheses were advanced: (1) Subjects shown contrasexual pictures would require more presentations to form a stimulus-bound percept than would subjects shown congruent pictures; (2) Extraverts would require more presentations of the contrasexual pictures to form a stimulus-bound percept than would introverts; (3) Extraverts would manifest more emotional distance when describing a stimulus bound contrasexual percept than would introverts. Hypotheses I and 3 received clear support, while Hypothesis 2 received indirect confirmation at early stages of percept development (prior to correct recognition). Results were discussed in relation to (l) Jung's theory of introversion-extraversion, focusing on content factors of the stimuli, and (2) Berlyne's arousal model, focusing on collative properties of the stimuli. Comparisons were drawn between the present microgenetic approach and a similar perceptual method, and between the present theoretical formulation and the concept of psychological androgyny. PMID- 16366901 TI - Data discrepancies in the Personal Orientation Inventory Manual. AB - The Personal Orientation Inventory Manual appears to contain a number of errors specifically related to the calculation of the scores for the time ratio and support ratio. Some are of a relatively minor nature, but several are important because they are directly related to the standards whereby individuals are classified as self-actualized, normal, or nonself-actualized. Although part of the discrepancy seems to be due to the number of questions left unanswered by subjects, this does not account for the total difference. At present, it is unclear why the inconsistencies exist. PMID- 16366902 TI - How and why people accept personality feedback. AB - The present experiment examined a model and a supplementary explanation of feedback acceptance. The model consisted of inventory responding under a yea saying response set and the explanation stated that feedback is accepted because it is accurate and persuasive. "Neurotic" and stable undergraduates (n = 120) took several personality inventories and then rated the accuracies of personality "interpretations." As predicted, the model mirrored every detail of feedback acceptance; also, the accuracy of interpretations related most simply to their acceptance and the undergraduates changed their self-perceptions in the direction of the interpretations. Contrary to predictions, individual differences in persuasibility did influence feedback acceptance. PMID- 16366912 TI - But it's only an inkblot. AB - The nature of the Rorschach is discussed using data from three temporal consistency studies with children of different ages, plus two other studies in which the retest reliability question was approached by experimental manipulation. It is postulated that the test response reflects the basic psychological styles or response tendencies that have developed within the personality, plus representations of the more private world of the person. The challenge for interpretation and research is to differentiate which of the test elements reflect each of these features, and to determine how they merge to create the response during a very brief time interval. PMID- 16366913 TI - Adaptive regression of two types. AB - Adaptive regression is considered to be of two types: a relatively uncommunicable private type and a relatively communicable public type. Private adaptive regression was operationalized by hypnotic suggestibility and by a questionnaire. Public adaptive regression was operationalized by two measures of esthetic sensitivity. It was hypothesized that the psychodynamics of the two types are as follows: Hypothesis 1: Private adaptive regression involves engaging in primary process because of secondary process motivation. Hypothesis 2: Public adaptive regression involves engaging in primary process because of secondary process motivation. Hypothesis 3: Public adaptive regression involves secondary process shaping of primary process to a greater extent than does private adaptive regression. Measures of engaging in primary process were derived from inkblot responses. Hypotheses 1 and 3 were supported: Hypothesis 2 was not. PMID- 16366914 TI - A note on the use of the Hand Test in cross-cultural research: comparison of Iranian, Chinese and American students. PMID- 16366915 TI - The development of sexual identity. AB - The development of sexual identity was studied in 686 individuals, using a fantasy measure of sexual identity that does not depend on learned sex-role stereotypes. The results showed a marked differentiation between the sexes at pre adolescence, a loss of differentiation during the freshman year of college that most closely resembled the lack of differentiation in sexual identity of preschool children, and a reemergence of a clear sexual identity during the last years of college. Possible biological, social context, and psychodynamic maturational explanations for this pattern of development are discussed. PMID- 16366916 TI - Ego development and sociometrically evaluated maturity: an investigation of the validity of the Washington University Sentence Completion Test of Ego Development. AB - The reported study was an attempt to validate the Washington University Sentence Completion Test of Ego Development (WU-SCT), using a broad, sociometric index of maturity as the criterion variable. Sixty college women living in a sorority house completed the WU-SCT and evaluated one another's readiness for mature functioning in each of four adult social roles: career, marriage, parenthood, and community involvement. The reliabilities and the intercorrelations of the ratings suggested that peers distinguished two facets of maturity. While results support the hypothesized relationship between WU-SCT scores and a global sociometric index of maturity, the test's substantial correlations with indices relating to relatively impersonal roles (career and community) and apparent lack of correlation with indices relating to intimate interpersonal roles (marital and parental) suggests that the validity of the WU-SCT may be less broad than Loevinger's theory would imply. Several alternative interpretations of the data are suggested. PMID- 16366917 TI - Physical attractiveness, romantic love, and equity restoration in dating relationships. AB - Measures of physical attractiveness, romantic love, and dominance were given to a sample of 123 dating couples. Contrary to expectation, attractive subjects were not loved more than those judged as less attractive. As suggested by equity theory, however, subjects who believed that their partners were the more attractive member of the dyad loved their partners more (p < .05) and indicated greater submission in their relationships (p < .01) than those who believed that they were the more attractive member. The results suggest that as the dating relationship progresses, the relative difference in attractiveness between partners may become a more important determinant of attraction than overall level of attractiveness. PMID- 16366918 TI - Relations between level of ego development and adjustment pattern in adolescents. AB - The present study was designed to assess the relation between Loevinger's hierarchy of ego development stages, measured by the Washington University Sentence Completion Test, and adjustment patterns as measured by the Minnesota Multiphasic Personality Inventory. A total of 150 high school students between the ages of 14 and 15, 75 boys and 75 girls, served as subjects. It was hypothesized that certain adjustment patterns were more prevalent at certain points of the ego development hierarchy than at others, specifically: (a) hypochondriasis and psychopathic deviance at the ego stages below conformity; (b) hysteria at the conformist ego level; and (c) obsessive-compulsiveness and paranoia at the ego stages above conformity. All of these hypotheses, except that regarding psychopathic deviance, were supported by the data. One unpredicted relationship, a tendency for depression to be most characteristic of the conformist ego group, was found. PMID- 16366919 TI - Use of local norms to improve high-point code-type concordance of two short forms of the MMPI. AB - Criticisms of short forms of the MMPI have been based on claims of unsatisfactory high-point code-type concordance rates. The use of local norms has been suggested as a possible solution for these low concordance rates. In this study local norms for three client populations were developed for the MMPI-168 and Faschingbauer's abbreviated MMPI. These local norms did not significantly increase high-point code-type concordance. Possible explanations for these findings are discussed and suggestions for future research offered. PMID- 16366920 TI - Conversion equations for modified scoring of the MMPI-168. AB - For a sample of 300 patients who had been administered the Minnesota Multiphasic Personality Inventory (MMPI), the MMPI-168 was extracted from the full MMPI and scored to incorporate those items normally excluded by Form R keys. MMPI-168 correlations with the full MMPI ranged from .80 to .97 with a mean of .90, indicating satisfactory statistical validity, and modified scoring was shown to improve predictability for Pa and Sc. Using these data, substitution equations for transforming MMPI-168 raw scores to estimates of full-scale scores were calculated. These transformations did not differ greatly from those reported in previous research except on Pa and Sc, where additional items increase scale length substantially. PMID- 16366933 TI - A Test-Operate-Test-Exit model for Rorschach scoring. PMID- 16366934 TI - Methodological, developmental, and sex biases in the Social Self-Esteem test? Modest, no, and no. AB - Data on the Social Self-Esteem (SSE) test were examined for two potential method artifacts, sex differences, and developmental effects. A single SSE item was administered to 2009 secondary school students as part of an in-class, self administered questionnaire. Across four orders of presentation, the measure performed according to construct-derived predictions. Modest order effects emerged when the conditions were examined separately. No evidence for random response was found. No grade by sex interaction was found. Separate trends for grade and sex effects were weak and contrary to the predictions derived from an earlier critique of the measure. It was suggested that use of the SSE be limited to research applications aggregating individuals' responses. PMID- 16366935 TI - Independent CPI masculinity and femininity scales: psychological correlates and a sex-role typology. AB - The meanings of masculinity and femininity as measured by independent scales (MSC and FMN) on the CPi are explored, and a sex-role typology is constructed. The CPI and ACL were administered to 287 college students. Correlations of MSC and FMN with 18 CPI measures showed both masculinity and femininity to be positive traits. Masculinity included self-confidence, mental alertness, assertiveness, leadership, and poise; femininity included socialization, self-control, development of ethical standards, and emotional sensitivity. A sex-role typology was formed, based on joint MSC and FMN scores, and each sex-role type was described, based on its relationship with 18 measures on the CPI and 300 adjectives on the ACL. The typology showed that the high MSC/high FMN group described themselves in favorable terms. The low MSC/low FMN group appeared unhappy, somewhat withdrawn, and distrustful. The self-description of high MSC/low FMN and low MSC/high FMN groups corresponded to stereotypic views of masculinity and femininity, respectively. The similarity of these findings with research using different approaches suggests that consistent pictures of masculinity, femininity, and sex-role types are emerging. PMID- 16366936 TI - Equating the social desirability of Bem Sex-Role Inventory masculinity and femininity subscales. AB - Two experiments investigated the social desirability of Bem Sex-Role Inventory (BSRI) items. Mean social desirability of the masculinity subscale was significantly higher than that of the femininity subscale in both experiments. A revision of the BSRI femininity subscale was suggested which replaced three items of questionable social value with three more socially desirable items and virtually eliminated the social desirability difference between masculinity and femininity subscales. PMID- 16366937 TI - Absorption in imaginings, sex-role orientation, and the recall of dreams by males and females. AB - Questionnaire measures of dream recall frequency, number of sensory qualities in dreams and emotion in dreams were correlated with several imaginal ability and personality variables in male and female college students. Absorption (i.e., subjects' degree of involvement in such activities as daydreaming, watching a movie, etc.) was the most important predictor of dream variables in females. In males sex-role orientation contrary to stereotype (i.e., femininity) was the only variable significantly related to a dream variable (i.e., frequency of dream recall). The implications of these sex differences for research on dream recall are discussed. PMID- 16366938 TI - Item format and the structure of the Eysenck Personality Inventory: a replication. AB - A seven-choice response format for personality inventories allows finer distinctions by subjects than the traditional two-choice format. The Eysenck Personality Inventory was employed in the present study to test the hypothesis that use of the expanded format would result in a clearer indication of test structure. A principal components analysis with a varimax rotation retained two components for the two-choice format, clearly identifiable as Eysenck's "Neuroticism" and "Extraversion" which accounted for 16% of the variance. The expanded format retained four components accounting for 28% of the variance. PMID- 16366939 TI - Birth order, family structure and personality variables. AB - The birth order and family structure variables assessed included first or only child, position in multi-child family, position in dyadic family, age of parents at birth of given child, whether reared by one or both parents, whether reared under permissive or authoritarian family atmosphere. The personality variables assessed included Sociability, Anxiety, Dominance, Superego, Phlegmatic Temperament, Involvement, Self-Regard and other scales measured by the Howarth Personality Questionnaire. One hundred seventy female and 142 male undergraduates were tested. Significant relationships found included: anxiety lower in firstborns; superego higher in firstborns; persistence higher in those not dominated by elder sibling; phlegmatic temperament higher in permissive families; involvement and phlegmatic temperament lower in single parent families; self pride higher in the elder of a sibling pair; individual tolerance higher in children reared by both rather than single parent. PMID- 16366940 TI - Inventory responding models people's acceptance of feedback 'derived' from tests and from interviews. AB - Sixty-four undergraduates were individually and randomly given one of four assessment procedures: The Rorschach, Minnesota Multiphasic Personality Inventory (MMPI), Structured Interview, or Unstructured Interview. Students then completed a series of 7-point Likert-type rating scales indicating their perception of the assessment. One week later students individually received identical personality descriptors, presented either as personality "feedback" or as a personality "test". Finally, students completed a second Likert-type rating task, again indicating their faith in the assessment and perception of the examiner's skill. Layne and Michels' model that inventory responding parallels people's acceptance of Barnum feedback was supported. Descriptor favorability significantly enhanced acceptance of both "feedback" and inventory items, while type of prior assessment had no significant effect on either behavior. Student perceptions of assessment accurately paralleled characteristics of each type of assessment. Finally, student faith in the assessment and perception of the examiner's skill failed to significantly increase from before to after receiving "feedback" It was concluded that inventory responding accurately models the Barnum Effect and that people accept feedback rationally, not gullibly. PMID- 16366941 TI - The wit: a personality analysis. AB - Fifty-eight undergraduate psychology students were group tested on 12 variables thought to be related to the successful prediction of wit. In addition to a score on wit, scores on creativity, verbal fluency, need for play, need for impulsivity, self-confidence, dominance, exhibition, change, autonomy, personal adjustment, authoritarianism, and sex group were obtained. The results were analyzed in a stepwise multiple regression analysis. A significant Pearson product-moment correlation was obtained between wit and creativity (p < .01) and between wit and personal adjustment (p < .05). The multiple regression analysis indicated that creativity was the best single significant predictor of wit. Personal adjustment, in combination with creativity, provided the next highest significant contribution. The results are discussed and suggestions for further research are posited. PMID- 16366942 TI - A commentary on Adolf Eichmann's Rorschach. AB - A Rorschach was administered to Adolf Eichmann in Jerusalem in 1961 at the time of his trial for war crimes. The recent publication of this protocol offers an opportunity to compare his personal world against opinions formed about him by observers at his trial. Various professionals certified Eichmann as a banal, ordinary man, and a societal theory was proposed about Nazism based in part on impressions of Eichmann as an uncomplicated man. This commentary examines Eichmann's protocol and provides an opinion that in several important respects his record includes features uncharacteristic of an ordinary, banal mind. PMID- 16366956 TI - The assessment of Transactional Analysis ego states via the Adjective Checklist. AB - Transactional Analysis (TA) conceptualizes personality in terms of five functional ego states: Critical Parent (CP), Nurturing Parent (NP), Adult (A), Free Child (FC), and Adapted Child (AC). The present investigation was designed to develop a procedure for the assessment of the relative strength of ego states from the responses of persons who are not familiar with TA theory. Fifteen TA experts assigned ego-state ratings to the 300 items of the Adjective Check List (ACL) with high interjudge agreement being found. Using the judges' mean raft rigs, a system was developed for obtaining ego-state scores for any selected set of ACL adjectives. The use of the system was illustrated by its application to the ACL self-descriptions of 100 university students which were analyzed in terms of "egograms" in which the five functional ego states were expressed relative to one another. A test-retest study involving an additional 154 students demonstrated stable individual differences in ego-state scores. Mean egograms of female and male students did not differ from one another but showed appreciable differences from the expert judges' ratings of the "psychologically healthy person." The potential Usefulness of the procedure in clinical practice and in testing TA theory is discussed. PMID- 16366957 TI - Development of the Mental Health Locus of Control and Mental Health Locus of Origin Scales. AB - The Mental Health Locus of Control (MHLC) Scale is an area-specific measure of locus of control expectancies designed to predict mental health related behaviors, particularly those occurring in treatment situations. The discriminant validity of the MHLC, in contrast with Rotter's 1-E measure of generalized expectancies, was demonstrated with two mental health related variables: beliefs concerning etiology of psychopathology, and information about abnormal psychology. Beliefs about etiology were measured by the Mental Health Locus of Origin (MHLO) Scale. The primary hypothesis, that individuals with 'endogenous' beliefs about etiology tend to have 'external' expectations for the client's role in treatment, was supported by a statistically significant (p < .001) correlation between the MHLC and MHLO Scales. PMID- 16366958 TI - Scales measuring people's perception of themselves as origins and pawns. AB - Scales measuring people's perception of themselves as origins (determining their own behavior) or pawns (their behavior being determined by forces beyond their control) were devised using content analysis of verbalizations. Scoring categories were based on the work of Heider (1955). Reliability was satisfactory. Normative data were obtained from 528 people, each experiencing one of nine life situations. Pawn and origin perception differed across situations and these differences were correlated with independent ratings of how people would perceive the situations. Pawn perception was more stable over time than origin perception but both varied over time. Origin scores were relatively independent of measures of negative affect. Pawn scores were associated with hostility directed inward, ambivalent hostility and total anxiety. Associations were found with some forms of Coping behavior and people's reactions to a relative's serious or less serious illness. Origin and pawn scores were not negatively correlated, indicating the inappropriateness of regarding them as opposite ends of a single dimension of causal perception. PMID- 16366971 TI - A comparison of the Bem Sex-Role Inventory and the Heilbrun Masculinity and Femininity Scales. AB - This study compares two instruments which have recently been devised to measure sex-role identification, Heilbrun's Masculinity and Femininity Scales and the Bem Sex-Role Inventory. Correlations between the masculine and feminine scales of these instruments were significant for male but not female subjects; intrascale comparisons found no relationship between the Bem scales but moderate correlations between the Heilbrun scales for male subjects. There was agreement between the two measures in classifying approximately 47% of the subjects into one of the four sex-role categories. Misclassification occurred primarily on categories which have been found to show considerable overlap in personality characteristics. PMID- 16366972 TI - Parental locus of control and the assessment of children's personality characteristics. AB - The relationships between different parental locus of control orientations and their children's personality characteristics of locus of control, anxiety, intelligence, achievement, and behavioral adjustment were examined for 134 fourth grade children and their parents. Correlational results showed that mothers' locus of control was more Consistently related to their children's Characteristics than was the fathers' locus of control although this depended somewhat on the sex of the children. Further analyses revealed that parents who were both high in external locus of control had children who scored high in anxiety and low in intelligence. However, sets of parents in which at least one of the spouses was internally controlled had children who did not differ in their personality characteristics. Results indicate that further investigative research is warrented on how parenting attitudes and behaviors affect the development of such personality characteristics in children. PMID- 16366973 TI - The relationship of the Marlowe-Crowne Scale and its components to defensive preferences. AB - Two lines of recent research have interpreted the Marlowe-Crowne Social Desirability Scale (MC-SDS) as a measure of either need to avoid social disapproval or need to protect and maintain vulnerable self-esteem. In addition, it has been suggested that two component (Attribution and Denial) scores derived from the MC-SDS are differentially related to defensiveness. The present study tested several hypotheses regarding the relationship of the MC-SDS and its components to relative preference for five classes of defensive responses, as measured by the Defense Mechanisms Inventory. Results among females best supported a disapproval-avoidance interpretation of the MC-SDS. while no support was found for the proposed differential validity of the attribution, and denial components. Support for such differential validity was, however, obtained among males. Results for this group suggest that high denial component scorers will opt for protecting vulnerable self-esteem even in the face of possible social disapproval, while high attribution scorers are primarily concerned with avoiding such disapproval. Implications for use of the MCSDS and its components as measures of defensiveness are discussed and suggestions for additional research are presented. PMID- 16366974 TI - Individual differences in Pollyannaism. AB - The Pollyanna Principle states that people process pleasant information more accurately and efficiently than less pleasant information. This study examined whether different measures of Pollyanna tendencies are correlated with each other. Fourteen measures of Pollyannaism were derived for 133 students. The results showed that subjects who rated themselves as optimistic or happy also showed Pollyannaism on other measures of happiness, believed that the events in their lives were pleasant, gave themselves positive ratings on personality characteristics, recalled pleasant words more often than unpleasant words, supplied more free associations to pleasant stimuli than to unpleasant stimuli, listed pleasant items first, and judged that pleasant words were more frequent in the English language. PMID- 16366975 TI - Personality correlates and sex differences in the topological representations of self and others. AB - This investigation attempted to identify the underlying components of the Social Self-Esteem measure. Although devised to function as a brief nonverbal measure of self-esteem applicable to a wide range of special populations, understanding of the measure remained limited. Relationships of this measure to related personality constructs and socioeconomic variables were examined to develop a clearer picture of what the Social Self-Esteem measure is actually assessing. Results indicated that the personality factor Capacity for Status was significantly related to the measure's assessment only among males. it was suggested that the Social Self-Esteem measure assesses some aspect Of what may be termed an individual's status ideal. The usefulness of differentiating scoring levels on the measure is demonstrated. Reasons for the lack of measurement usefulness among females were discussed, in terms of continued reliance among females upon significant others for status satisfaction. PMID- 16366976 TI - Another look at personal validation. AB - Students' ability to validate the results from their own psychological tests was examined in two experiments. In Experiment I, sophomore, senior, and graduate students were presented with their actual and inverted profiles from the California Psychological Inventory (CPI) and the Students were asked to select their actual profiles. The senior and graduate students could reliably select their own profiles while the sophomore students could not. In Experiment 2, sophomore students were presented with their actual and inverted profiles from the CPI and the Differential Aptitudes Test (DAT) and the students were asked to select their actual profiles. The students were able to reliably select their actual profiles for both tests. The students were much more likely to select their own DAT profile than CPI profile, and they were more confident in their selection of the actual DAT profile. PMID- 16366990 TI - Paul E. Meehl: Distinguished Contribution Award in New York, September 2, 1979. PMID- 16366977 TI - The common space of personality traits and vocational interests. AB - Investigated relationships between personality traits and vocational interests. Cattell's (Research and Consultations Center of Educational Personnel, 1976) CAQ Part I and the Ramak interest inventory (Meir, 1975) based upon Roe's (1956) occupational classification system were administered to a sample of 397 university applicants. Canonical Correlation Analysis and Smallest Space Analysis were used to test two hypotheses (a) relationships exist between personality traits and vocational interests; and (b) personality traits which characterize occupational profiles are arranged in circular order corresponding to the configuration Of the vocational fields which represent those occupations. Both hypotheses were supported by the data. PMID- 16366991 TI - Personality characteristics of androgynous and sex-typed females. AB - Personality characteristics of androgynous and sex-typed females identified by the gem Sex-Role Inventory were studied using the California Psychological Inventory. The results indicated that sex-typed females showed personality characteristics associated with traditional sex-role stereotypes while androgynous females showed both masculine nd feminine characteristics. There was an indication that androgynous and masculine females were better adjusted socially than feminine or undifferentiated females. PMID- 16366992 TI - Stability of results across many studies: sex differences on the personal orientation inventory. AB - An analysis of sex differences on the Personal Orientation Inventory (POI) was used to illustrate a method to assess the stability of results across many studies. It was argued that sex should not be Used as a moderator as significant differences between males and females were not found consistently across six studies using the POI. PMID- 16366993 TI - Is the acquiescent response style problem not so mythical after all? Some results from a successful balanced F scale. AB - It is shown that Rorer's exoneration of the F scale from acquiescent response style contamination is dependent on the finding-that various acquiescence measures fail to intercorrelate. When acquiescence is measured as the total score on adequate balanced scales scored without reversals, significant internal reliability is found. It is found, in fact, even with scales that are not particularly ambiguous. It is concluded that some scales are not responded to meaningfully by some people and if these people are not to be confounded with real high scores, balancing against acquiescence is still needed. PMID- 16366994 TI - Checking the reliability and validity of HSPQ profiles. AB - Developed a scale designed to detect invalid response patterns on the High School Personality Questionnaire (HSPQ). Using a large sample of protocols drawn from the HSPQ standardization group, ten items were found to satisfy the necessary criterion for inclusion in the resulting "random" or "validity" (RV) scale. When applied to separate cross-validation groups of nearly 2,000 valid and randomly generated response protocols, RV was found to correctly classify 79% of the profiles. As a further test of the scale's utility, 100 profiles classified as valid and 100 classified as invalid were selected for comparison. In the invalid group, KR-20 reliabilities of the 14 trait scales were almost uniformly lowered, Also, for the invalid group the correlations among the trait scales were attenuated and altered to the extent that the underlying factor structure of the test was upset. PMID- 16367006 TI - Some old and new directions in Rorschach testing. AB - The perennial dispute between clinicians and researchers as to the validity of the Rorschach is discussed. It is suggested both groups are correct in that the Rorschach provides both poor psychometric data as seen by the researcher and rich idiographic data as seen by the clinician. The freedom of the Rorschach response process is viewed as largely responsible for both these situations. Some possible new directions for Rorschach usage and research are discussed which may improve on the quality of the idiographic information provided by the Rorschach test. PMID- 16367007 TI - The Mosher Guilt Scales: a construct validity extension. AB - The three subscales of the Mosher Guilt Scales (sex guilt, hostility guilt, and morality conscience guilt) and several personality and attitude measures were ad ministered to college subjects in an effort to examine the construct validity of the Mosher Guilt Scales. The variables that were compared with the guilt subscales were hostility, anxiety, religious orthodoxy, self-esteem, and the importance of religious and economic values in one's life. The results indicated that both the construct validity of the Mosher Guilt Scales and the need for the continued use of its three subscales were supported. Among the most crucial findings of the study were that both of the religiosity measures were positively correlated with all three guilt subscales, and that economic values were negatively correlated with both sex guilt and morality-conscience guilt. PMID- 16367008 TI - Personality characteristics associated with aerobic exercise in adult males. AB - Using an index of aerobic conditioning 23 adult male exercisers and 23 nonexercisers were identified. During individual interviews with each subject basic demographic data were obtained and blood pressure, resting pulse rate, and body fat were measured. All subjects completed Form A of the Sixteen Personality Factor Questionnaire (16PF). Statistically significant differences occurred on six primary factors and four Secondary dimensions of the 16PF. In comparison to the inactive subjects the exercisers, all of whom were joggers or runners, were more reserved, expedient, suspicious, forthright, liberal, and self-sufficient on the primary traits, and were more alert and independent, less discreet, and evidenced lower super-ego strength on the broader secondary patterns. The statistically significant training effects were lower pulse rate and less body fat, but did not include blood pressure. PMID- 16367009 TI - A comparison of statistical infrequency and subjective judgment as criteria in the measurement of originality. AB - The Alternate Uses test was administered to a sample of 60 college students. Three different criteria of originality were used in scoring: subjective judgment, statistical infrequency, and random numbers. Correlations among the three methods exceeded .67 (p < .01). The explanation for this finding is that the originality scores are strongly influenced by the number of responses (i.e., ideational fluency). Because the total originality score is based on a sum of the originality weightings for each response, individuals who give many responses are more likely to get high originality scores. Further analysis demonstrated that the subjective and statistical infrequency scoring techniques are not correlated when ideational fluency is controlled. Implications for the measurement of originality are discussed. PMID- 16367023 TI - Are mood states bipolar? AB - Aims were (a) to test whether or not mood state factors are bipolar when response styles are controlled, and (b) to test whether the mood states conform to a circular order Intensity ratings were collected from 204 college students on a 63 adjective scale defining nine moods. Another sample of 127 rated their moods on modified four-category certainty scales. Factor analyses of the 63 adjective correlations disclosed seven monopolar factors. When acquiscence was controlled, the analysis yielded four bipolar and one monopolar mood states. The analyses of the data collected on the modified ratings form, which minimized response bias, disclosed three bipolar moods (composed-anxious, energetic-tired, agreeable grouchy) and two monopolar states (cheerful and dejected). The hypothesis of a circular order of mood states received some support. PMID- 16367024 TI - The measurement of locus of control in black preschool and primary school children. AB - Developed a form of the Preschool and Primary Nowicki-Strickland locus of control scale (PPNSIE) for use with black children (PPNSIE-B). The psychometric data suggest that the new scale is worthy of further examination and development. PMID- 16367025 TI - The universality of generalized personality statements. AB - Researchers for three decades have assumed that generalized personality statements are nearly universally valid. In this study, undergraduate students indicated whether generalized personality statements were "true" or "false" as applied to themselves; then these same students and a group of graduate students estimated the percentage of college students who would answer each generalized statement "true." Only 7 of the 12 commonly-used, generalized personality statements were found to be nearly universally valid among the undergraduates; the other 5 were answered "true" by fewer than two-thirds of the undergraduates. Both groups undergraduates and graduate students were very accurate in estimating the percentage of students who would respond "true" to each statement. Thus, many supposed generalized statements are not nearly universally valid, and students are cognizant of the degree of universality of generalized statements. Results of prior research based on inappropriate assumptions about generalized statements must be viewed cautiously. PMID- 16367026 TI - Runners and meditators: a comparison of personality profiles. AB - Although both running and meditating have been considered as healthful and enlightening, no data have yet been published comparing the personality profiles of the two groups. A sample of 48 runners and 43 meditators, all males between the ages of 40 and 60, were given the Cattell 16 PF Test Profile and the results were compared. The meditators were significantly more assertive and enthusiastic than the runners. They also appear as Significantly less conscientious and controlled than the runners, as well as more experimenting and suspicious. PMID- 16367027 TI - Personality characteristics associated with aerobic exercise in adult females. AB - Using an index of aerobic conditioning 27 adult female joggers and 25 nonexercisers were identified. During individual interviews with each subject basic demographic data were obtained and blood pressure, vital capacity, resting pulse rate, and body fat were meaSured. All subjects completed Form A of the Sixteen Personality Factor Questionnaire (16PF). Statistically significant differences occurred on two primary factors, and one secondary dimension of the 16PF. The probability of three marginally significant differences out of 24 comparisons is well within the range of expected chance fluctuation. Thus, the study provided no evidence that personality characteristics are predisposing factors in the adoption of an aerobic jogging program by young adult females. The only statistically significant training effect was lower pulse rate; blood pressure, vital Capacity, and body fat were similar for the two groups. Finally, the results of this study of females were compared to those of a previous investigation of male joggers. PMID- 16367028 TI - Inventory responding as a model of people's acceptance of personality interpretations. AB - The hypothesis was tested that inventory responding modeled the acceptance of personality feedback. A barnum group (n=40) was asked to rate the personal accuracies of a list of personality inventory items and then an equivalent list of bogus personality feedback. The two lines of evidence which supported the model were (a) that the correlation between their inventory ratings and their feedback ratings was not only Significant (p < .001) but achieved a ceiling magnitude, as evidenced by a control group's (n = 40) data; and (b) that the variables influencing, inventory responding exerted an equal influence upon feedback acceptance, Contrary to the model, it was found that feedback was accepted more highly than were the inventory items. Conclusions were that inventory responding, as a model of feedback acceptance, is accurate with respect to individual differences but somewhat inaccurate with respect to overall levels of acceptance due to the additional influences of persuasion. It was also concluded that clients' acceptance or rejection of personality feedback is not evidence of the validity or invalidity, respectively, of assessment devices. A reformulated model of personality feedback acceptance was proposed. PMID- 16367029 TI - Ambiversion: characteristics of midrange responders on the Introversion Extraversion continuum. AB - Sixty-nine midrange responders on the Introversion-Extroversion (IE) scale of the Eysenck Personality Questionnaire (EPQ) were given an experimental measure to assess tendency toward ambivalence (i.e., both strongly introverted and strongly extraverted responses) vs. moderation (i.e., midrange responses), and degree of importance given to IE types of activities. The participants were divided into those showing ambivalence vs. those showing moderate responses, and into those reporting high vs. low importance of such activities. As was predicted, the ambivalent group showed significantly higher scores on the neuroticism scale of the EPQ, while the low importance group tended to show more psychoticism, but not to a significant degree. Both of the midrange scores were unrelated to the Bem Sex Role Inventory. Based on the findings, at least two groups are posited in the midrange of the IE dimension: an ambivalent group with mixed strong introversive and extraversive tendencies, and an ambiverted group with midrange scores. PMID- 16367030 TI - Defensive style and performance on objective personality measures. AB - Two studies on the relations between various styles of defense and the power or validity of self-reports on objective personality questionnaires. The power of individual items on the Rotter Internal-External Locus of Control scale to discriminate between subjects defined as internals or externals by a median-cut procedure using the total score was not affected by the presence Or absence of a projective defensive style. However, the presence of repressive tendencies did appear to restrict item power. Prediction from two achievement scales taken from Gough's California Psychological Inventory was not influenced by level of repression, was affected somewhat adversely by a projective style, and was most clearly contaminated by a rationalizing defense. It was proposed that the differential effects of defensive styles upon the validity of personality tests depended upon the convergence or divergence of the cognitive operations involved in the type of defense and in the type of personality measure. PMID- 16367032 TI - The Kramer-Froehle controversy: a contribution to construct validity of the FIRO B questionnaire. AB - Replicated the conditions established by Kramer in his attempt to contribute to the construct validation of the FIRO-B. Froehle's apparent later replication produced significantly different results from Kramer's original study. In replicating Kramer's design this researcher wished to establish whether the earlier failure to replicate was due to a difference in design or to an actual lack of construct validity of the FIRO-B. Kramer's findings were supported and an alternative explanation for the difference in Kramer's and Froehle's findings is discussed. PMID- 16367031 TI - Internal control, powerful others, and chance: a confirmation of Levenson's factor structure. AB - The Internal-External control scales of Rotter and Levenson, and the Social Desirability scale of Marlowe and Crowne were completed by 71 male and 85 female students. Factor analysis clearly confirmed the three-factor structure underlying Levenson's questionnaire, while further analyses revealed close agreement with her findings on scale characteristics and interrelationships. Some new evidence for the multidimensionality of Rotter's scale was also presented. PMID- 16367043 TI - Two kinds of radicals: a discriminant analysis of a projective test. AB - Rorschach tests were administered to 36 subjects grouped according to ethnicity and political ideology. Pairwise discriminant analyses of scores significantly differentiated both Jewish and non-Jewish radicals from nonradicals in a manner consistent with previous survey findings. PMID- 16367044 TI - The assessment of ambitious-narcissistic character style with three projective tests: the Early Memories, TAT, and Rorschach. AB - This study describes the construction of scales designed to assess ambitious narcissistic character style on three projective tests: the Early Memories, TAT, and Rorschach. The main aim of the study was to evaluate the reliability and validity of these scales. A secondary aim was to demonstrate the feasibility of assessing particular character styles with projective tests commonly available to clinicians and researchers. Forty male college students volunteered as subjects. The overall reliability of the three projective scales was found to be acceptable. The scales were then tested for validity in two ways. First, they were intercorrelated and found to show a pattern suggesting a common dimensionality. Secondly, the scales were found to successfully differentiate subjects rated by clinically-trained raters as ambitious-narcissistic in style from those subjects rated as non-ambitious-narcissistic. PMID- 16367045 TI - 'Fear of success' imagery as a reaction to sex-role inappropriate behavior. AB - Cue effects on TAT imagery, often considered evidence of fear of success, were examined. The cue generally used to assess this motive (Horner, 1968) was modified to depict success in an occupation traditionally considered appropriate for females (ballet). This new cue and Horner's original "medical student"cue produced very different results in fear of success imagery, the modified cue eliciting significantly less of this imagery from females and more from males. A substantial portion of what has been interpreted as "fear of success" imagery appears to involve anxiety about engaging in achievement activities that have traditionally been considered inappropriate to one's sex-role. The implications of this finding for female achievement conflict are discussed. PMID- 16367046 TI - Relationships among projective and direct verbal measures of achievement motivation. AB - The present study was undertaken to explore ways projective arid nonprojective verbal measures of achievement motivation elicit comparable and dissimilar responses in a "Third World" sample of entrepreneurs as a test of McClelland's theory of economic growth. Results were generally consistent with the theory of achievement motivation, and suggested that high scores on n Ach were associated with high rates of industrial and agricultural output; low scores were associated with static or declining business. TAT measurement was the best predictor in that it correlated 0.68 and 0.48 with agricultural and industrial production. Results further lend support to past findings: namely, that Various achievement measures appear to be measuring dissimilar constructs. PMID- 16367047 TI - Factor structure of Rabin's Child Study Inventory. AB - Factor analysis of responses by 213 persons to Rabin's Child Study Inventory result in five reliable factors. These five, termed Parental Fatalism, Parental Nurturance, Children's Independence, Parental Instrumentalism, and Children's Happiness, are similar to Rabin's original four parental motives of Altruism, Fatalism, Narcissism, and Instrumentalism. Two basic dimensions by which the five factors may be characterized are (a) whether the parent or child is the object, and (b) whether the feeling is personal or impersonal. PMID- 16367048 TI - Measures of motivation. AB - Tested four new scales and replicated earlier findings of the Orientation and Motivation Inventory with 159 boys and 166 girls in four grades of a high school. Each of the 12 scales was subdivided for analyses into two halves. The intercorrelations among the 24 scale halves were analyzed by the method of principal components to identify 11 factors for boys and 10 for girls. Ten of the twelve postulated dimensions were confirmed in the varimax solution for each sex. A factor analysis of the correlations among the 12 scale scores disclosed five similar second order dimensions for boys and for girls. These were hypothesized to reflect Status Seeking, Belief in own Potency, Seeking and Helping People, Theoretical Interests. and Risk-Taking preference. PMID- 16367049 TI - Three aspects of self-disclosure as they relate to quality of adjustment. AB - Subjects completed two measures of actual self-disclosure (one for intimacy of self-presentation and one for inwardness of self-presentation)and a self-report of past self-disclosure. and California Psychological Inventory. For both sexes the two measures of actual disclosure correlated positively but neither instrument related positively to the self-report measure. Females disclosed more than males on both measures of actual disclosure. Quadratic, inverted-U relationships were hypothesized for the self-disclosure and CPI scales, but by and large they were not found. Linear associations were noted, the major ones being a positive pattern of correlations between the inwardness of actual disclosure and measures of flexible autonomy, responsibility and socialization for the males, a negative set of relations between the intimacy of actual disclosure and scales indicative of good interpersonal adjustment for the females, and a positive relationship between the self-report measure and scales indicative of social poise, extraversion and socialization for both sexes. PMID- 16367063 TI - Teaching psychological assessment: training issues and teaching approaches. AB - Clinical psychology graduate programs need to pay attention to important issues involved in teaching psychological assessment, if graduate students are to be adequately prepared. Recent studies have suggested a gap between academic training in psychological assessment and internship expectations. Graduate students are not as well prepared as many internship settings would like. In addition to learning the fundamentals, students need help in dealing with issues of categorizing individuals, using tests in helpful ways, taking responsibility for decision-making, and developing a balance between critical evaluation of tests and appreciation of their usefulness. Teaching approaches for dealing with these issues are discussed. The importance of academic and clinical agencies working together is stressed. Goals for graduate training in assessment are suggested. PMID- 16367064 TI - Personality correlates of Rorschach scoring determinants: hypotheses derived from structural analysis. AB - A paradigm was presented whereby the meaning of some major Rorschach scoring determinants could be explicated in terms of a classification system based on Structural Analysis which indicates whether a response is intellectually or emotionally oriented and where it is located on a behavior-fantasy continuum. Implications of this analysis for resolving some interpretive issues and generating behavioral predictions were discussed. PMID- 16367065 TI - The temporal stability of some Rorschach features. AB - One hundred nonpatient adults, screened for evidence of personality disorganization, were retested after a three-year interval to study the temporal consistency of the Rorschach. In general, the correlational analysis for 19 basic variables studied, and a directionality analysis for five ratios, illustrate a considerable sturdiness over time. It is postulated that these variables can be separated into situational related indices (state variables), and more stable scores indicative of durable response styles (trait variables), based on the assumption that variables with lower correlations would identify state variables, while the higher correlations would signify the trait features. Nine of the 19 variables yield retest correlations in excess of. PMID- 16367066 TI - An investigation of the reliability of the Rosenzweig Picture-Frustration (P-F) Study, Children's Form. AB - Presents data on the retest and split-half reliability of the Rosenzweig Picture Frustration (P-F) Study, Children's Form, for two groups of subjects (aged 10-11 and 12-13 years, respectively), each group tested twice at an interval of three months. It was found that the retest reliability for all scoring categories (except O-D) were statistically significant (.01 or .05 level). GCR proved stable for the younger group (r = .53) but not for the older group. The difference here may be due to the less comfortable retest conditions prevailing for the older subjects. Reliability by retest was consistently higher than by the less appropriate split-half method, but the latter proved, nevertheless, to be in the main favorable. PMID- 16367067 TI - Quality of drawing as a factor in the interpretation of figure drawings. AB - Examined the drawings of 32 poliomyelitis patients and their matched controls to see whether figure drawings primarily reflect the subject's projection of psychological state, ability to draw, or some combination of these two factors. An overview of the literature is also given. Drawings from disabled and nondisabled subjects were reliably rated for quality, with no significant quality difference found between groups. Analyses of variance were then used to compare the drawings on several different measures of drawing size, completion and movement that might be assumed on the basis of the literature, to reflect the subjects' projection of disability status. Results showed that quality of drawing was a significant factor in 13 of the 17 comparisons while disability status proved to be a significant factor in only one of the 17 comparisons. There were no significant interactions. Therefore, the overall findings are consistent with the hypothesis that quality of drawing-rather than projective mechanisms-may at times be the overwhelming determinant of clinical and research findings with figure drawings. PMID- 16367069 TI - A note on "MMPI scale development methodology". AB - Clopton's recent article (1978) in JPA calls attention to methodological problems in the construction of special inventories from the MMPI pool, but overlooks the major problem. It stems from the theoretical correlation between psychopathology and the construct to be measured by the special inventory. This contamination renders the usual validity demonstration questionable. It is pointed out that there is a good number of special inventories that are not subject to the methodological problems noted by Clopton. PMID- 16367070 TI - Sex role conformity in homosexual and heterosexual males. AB - Nine principal components were extracted from the scores of 26 homosexual and 26 heterosexual males matched on age, education, occupation status, and sampling frame on Adjective Check List, Bem Sex Role Inventory, Chapin Social Insight Test, Experience Inventory, and Sixteen Personality Factor Questionnaire scales selected to represent a variety of "masculine" and "feminine" sex role attributes. The four most general varimax factors, accounting for 48% of the total variance, were: I) "Feminine Openness," II) "Masculine Boldness," III) "Reflection," and IV) "Novelty Seeking." Factor scores were constructed and indicated that the homosexual sample was significantly more identified with Factor I (p < .00l; 17% of total variance). There was no significant difference between the samples on Factor II, nor on the remaining two general factors which were not sex typed. PMID- 16367071 TI - Effects of three conditions of administration on Bem Sex Role Inventory scores. AB - In order to assess the validity of the technique employed in selecting items for the Bern Sex Role Inventory (BSRI) 58 male and 62 female undergraduates took the BSRI under three different sets of instructions. Both males and females were able to produce extremely masculine and extremely feminine BSRI profiles which differed (p < .01) from their own BSRI profiles. The results support Bem's (1974) assertion that the BSRI Masculinity and Femininity scales are comprised of items which are consistent with widely held sex-role sterotypes. PMID- 16367072 TI - Further evidence on 16 PF distortion scales. AB - Norms were obtained on a nationally representative sample of 4830 adult men and women for two empirically developed faking indices for Cattell's 16 Personality Factor Questionnaire (16PF) by Winder, Karson, and O'Dell (1975). These data provided convincing evidence that the cutoff suggested in the original work for the faking good scale was far too liberal and would routinely classify more than half of all 16 PF protocols as invalid. The faking bad cutoff appeared to be approximately correct. The correlations of the faking scales with the 16 PF primary factors were highly congruent with those reported in the development study and provided additional validity evidence for the two indices. Both indices were found to be sufficiently reliable to permit adjustments to be made in the primary trait scales when distortion is above average. PMID- 16367073 TI - The development of the Telic Dominance Scale. AB - The development of a 42-item measure of a number of personality features derived from the theory of psychological reversals is reported. These personality features are (a) the extent to which a person is serious-minded, (b) the extent to which a person plans ahead and organizes himself in the pursuit of goals, and (c) the extent to which a person seeks to avoid arousal. These features together are seen to constitute a personality trait which is described as telic dominance. Data concerning the test-retest reliability, criterion-related and construct validity of the Telic Dominance Scale are presented, together with the scale and scoring key. PMID- 16367074 TI - Objective personality assessment of children: an exploratory study of the Personality Inventory for Children (PIC) in a child psychiatric setting. AB - Some initial data are presented on the clinical utility of a new multidimensional objective personality inventory, the Personality Inventory for Children (PIC), Tabulation of symptoms and family characteristics independently derived from the chart data generated by 79 preadolescent outpatient evaluations resulted in an average of 12 correlates for each of the 16 profile scales. These correlates provided substantial support for the basic interpretive intent of at least 14 of these scales, as well as additional serendipitous construct validity in the form of PIC scale-parent descriptor relationships. The presentation of case findings of two adolescents with dissimilar PIC profiles demonstrated the potential value of this diagnostic instrument. PMID- 16367088 TI - Differential effects of ethnic membership, sex, and occupation on the California Psychological Inventory. AB - This study investigated the effects of race, sex, and occupation on college students' responses to the California Psychological Inventory. Analyses of variance of the standard CPI scales indicated numerous race, sex, and race X sex effects. A comparison of the results of factor analyses of the responses of black and white subjects (within each sex) indicated a similarity in factor structure between the two female groups, but not for the two male groups. These findings suggested that, due to these major differences in scale means and factor structure, care be taken in interpreting scores from the CPI when they are obtained from nonwhite populations. PMID- 16367089 TI - Sex differences and androgyny in fantasy content. AB - Fantasy stories were composed by 153 undergraduates (93 females, 60 males) who also responded to the Bem Sex Role Inventory. The fantasy stories were collected by group administration of a Thematic Apperception Test. The stories were content analyzed along 17 imagery categories. Males and females differed significantly in ten of these categories. The results indicate a substantial continuity of sex differences in fantasy content between earlier decades and the mid 1970s. However, women had relatively more imagery of a self-assertive, pleasurable, and careerist nature than had been found in earlier studies. The fantasies of sex typed persons were more situation-bound and more sexual than those of androgynous persons. Sex-typed persons appear to experience limitations in fantasy production which parallel their limitations in overt behavior. PMID- 16367090 TI - Irrational beliefs and personality. AB - This study investigated the relationship between irrational beliefs and the nonpathological personality characteristics origence and intellectence described by Welsh (1972, 1975b). The Adjective Check List (ACL) and the Irrational Beliefs Test (IBT) were administered to 319 college students. Data were analyzed for the entire sample and for males and females separately. In each case, two combinations of scores yielding significant canonical correlation coefficients were produced. The results offer a refinement to Ellis' personality model and suggest that irrational beliefs are insufficient as indications of psychopathology. PMID- 16367091 TI - The Mehrabian measures of achieving tendency: are separate male and female scales necessary? AB - The 1969 male and female Mehrabian measures of achieving tendency are among the best-known measures of resultant achievement motivation. This study addressed the question of whether unique male and female items are necessary. Pearson product moment correlation analyses on 83 male and 184 female undergraduates showed that females' scores on the unique male items correlated positively with their scores on the original female scale, and males' scores on the unique female items correlated positively with their scores on the original male scale. Coefficient alpha reliabilities of the original male and female forms were found to be acceptable. Cluster analyses showed that the Mehrabian items cluster in very similar ways for both males and females. PMID- 16367092 TI - Love, sex roles, and psychological health. AB - College students were administered a series of questionnaires designed to determine the association between psychological health, involvement in a romantic relationship, and the quality of love in a relationship. As predicted, subjects who had been involved in at least one love relationship scored significantly higher on a measure of self-actualization than individuals who had never been in love. In addition, it was found that level of self-actualization directly correlated with the degree of healthy love (Maslow's B-love) among individuals who had been involved in a romantic relationship. Among individuals who had terminated their relationship, those who demonstrated higher levels of self actualization felt less resentment toward their ex-lover. Furthermore it was discovered that females show a higher level of B-love than males, but contrary to predictions the length of a romantic relationship did not influence B-love. It is concluded that the results of this study are essentially consistent with Maslow's theories about self-actualization, hierarchy of needs, and healthy love. PMID- 16367093 TI - The Mosher sex guilt scale and the college population: a methodological note. PMID- 16367094 TI - The discriminative validity of the Whitaker Index of Schizophrenic Thinking. AB - An attempt was made to assess the discriminative validity of the WIST with a sample of psychiatric inpatients. Through the use of a standardized structured interview and a diagnostic system for schizophrenia based on the use of discriminant function analysis with nonpathognomic symptom combinations, a reliable and valid system was used to establish the criterion diagnosis. Approximately 63% of patients reliably diagnosed as schizophrenic were detected via the WIST while only 14% of nonschizophrenic patients scored in the schizophrenic range on the WIST. The assets and liabilities of using the WIST to diagnose schizophrenia are discussed. PMID- 16367107 TI - Individualized assessment and phenomenological psychology. AB - Although there is growing openness to tailoring of assessment procedures and reports to the particular client, these efforts typically have been sporadic and incomplete. This article reviews a systematic approach to individualized assessment, one whose practices are referred to as collaborative, contextual, and interventional. Clinical examples of these practices are presented in terms of their grounding in phenomenological psychology. Prior to that, themes such as intentionality, situatedness, dialectics, structuralism, and hermeneutics are introduced briefly. Phenomenological psychology as such is not seen here as necessary for all individualized practices, but it is seen as a critical touchpoint for development of theory and further practices. PMID- 16367108 TI - Dimensions of Picture Identification Test need associations. AB - To find a simple and more general structure for need association matrices obtained from Picture Identification Test (PIT) results, PIT need association matrices of eight college student groups were analyzed by INDSCAL, a multidimensional scaling technique. A common three dimensional structure was found. Based on the scale locations of the 22 needs, the dimensions were labeled the Combative, the Personal, and the Competitive dimensions. The INDSCAL model was used as a target for six other comparison groups including male and female English college students, male addicts and controls, and male delinquents and controls. There was enough similarity between the dimensional structure of the need associations of the comparison and target groups to assume that all groups shared the same basic structure of need associations. There were enough differences, however, to suggest that personality characteristics of groups may be related to deviations in the associative structure of their motivation systems. Applications of the techniques to clinical and personality evaluations of groups and individuals were discussed. PMID- 16367109 TI - Test-retest reliability of the Hand Test with normal subjects. AB - The Hand Test (Wagner, 1962) was administered to 71 subjects; 14 days later these subjects were again administered the Hand Test. Results indicated the Hand Test is a highly reliable measure of an individual's behavioral action tendencies. PMID- 16367110 TI - Acquiescence and the Bem Sex-Role Inventory. AB - The Bem Sex-Role Inventory (BSRI) and the Couch and Keniston Agreement Response Scale (ARS) were administered to 70 male and 72 female undergraduate students. Using the ARS scores as the dependent variable in a 2 (gender) X3 (sex-role identification) unweightedmeans factorial analysis of variance, the results showed that the two main effects and the interaction. effect were not statistically significant (F< I in each instance) indicating the lack of relationship between the BSRI and acquiescent responding. PMID- 16367111 TI - Repression-sensitization and nonverbal expressiveness. AB - Investigated the relationship of status on Byrne's repression-sensitization dimension to novel measures of nonverbal' expressiveness under stress and nonstress conditions, where subjects' perceptions of the purpose of the dependent measures were varied. Forty subjects were divided into sensitizer and repressor groups on the basis of Repression-Sensitization (R-S) scores. Subjects viewed either an anxiety-producing industrial accident film or a control film. Subjects were then told that the purpose of subsequent testing was either the assessment of creativity or of personality, as measured by a new projective test. The 2 X 2 X 2 factorial design enabled the investigation of predictions based on ego defensive interpretations of repression-sensitization and Lefcourt's "attitude towards emotionality" view. Although the study failed to provide strong support for either interpretation in full, the data were viewed as reconcilable with Lefcourt's theory where repressors, but not sensitizers, are-concerned. PMID- 16367123 TI - Measurement of delay of gratification. AB - To determine if psychoanalytic and social learning theorists are referring to the same phenomenon when they speak of "delay of gratification", 26 college undergraduates were divided into high and low groups on the basis of their scores on a "money-spending" questionnaire (a social learning measure). The difference between these two groups with respect to Rorschach M responses (the psychoanalytic measure) was nonsignificant. It was suggested that psychoanalytic and social learning theorists are probably referring to different phenomena when they speak of "delay of gratification." PMID- 16367124 TI - Personality differences between established and less-established male and female creative artists. AB - Past research on personality constructs of creative individuals has focused on men rather than women and has neglected the essentially social aspect of being considered creative. Therefore, personality variables of 60 male and 60 female creative artists were described using the California Psychological Inventory (CPI). Male and female groups were further divided equally into two subgroups reflecting a difference in degree of social acceptance of their work. Relationships between the four groups were explored by multiple discriminant analysis. The major conclusion noted was that creative females tend to be more like their creative male counterparts, in terms of CPI scale variation, than are males and females in general. PMID- 16367125 TI - Masculinity and femininity as factors in feminism. AB - The relationship between feminism and sex roles was explored in two studies. In female college students, sex-role types were measured by Baucom's (1976) MSC and FMN scales and by Bem's Sex-Role Inventory (Bem, 1974). Feminism was measured by the Attitude Toward Women Scale (AWS, Short Form) (Spence & Helmreich, 1972) and the Women's Liberation Scale (WLS) (Goldberg, 1976). Two groups of women were predicted to be more profeminist than others. (a) those scoring high on both masculinity and femininity and (b) those scoring high on masculinity and low on femininity. Results of the study supported both hypotheses using the AWS but not the WLS. In the second study, MSC and FMN served as the basis for forming sex role types, and feminism was measured by participation in NOW. Again, a large number of women high on masculinity and low on femininity were feminists: however, women high on both masculinity and femininity were underrepresented in NOW. Different results in the two studies are discussed in terms of the different ways that feminism was measured. PMID- 16367126 TI - Development and construct validation of a measure of attitudes toward public exposure to sexual stimuli. AB - Developed a brief, reliable, and valid measure of attitudes toward public exposure to sexual stimuli. Both advocates and opponents of public exposure to such stimuli typically cite presumed effects upon children. Consequently many of the Likert format items in the several versions of the scale deal with prescriptive and prescriptive beliefs about the exposure of children to sex related stimuli. Over a three-year period five different groups of respondents participated in the study. Both the longer and the shorter versions of the scale administered to these groups appear to have acceptable reliabilities. In an attempt to provide construct validation information, the relationships between the Acceptance of Public Sexuality Scale and several measures of traditionalism were examined. As expected, scores on the scale were inversely related to measures of traditionalism and positively related to measures of modernity Possible uses of the scale were discussed. PMID- 16367127 TI - A note on the actuarial interpretation of WAIS profile patterns. AB - The present study offers additional information on the actuarial interpretation of WAIS profile patterns. Specifically, a sample of 242 WAIS protocols was investigated for cases which met all the previously established classification criteria for a profile pattern called Cluster IV. Two cases were identified, and their unique relationships to previously delineated personality and behavioral descriptors were described. The results suggest that the Cluster IV profile pattern is stable and can be reliably associated with certain personality descriptors, and further, that age acts as a moderator variable which alters the interpretation of this pattern. PMID- 16367128 TI - The role of affective assessment in intelligence testing. AB - Based on the tenets of logical learning theory and the research in its support, it was predicted that subjects (87 seventh and eighth graders) would score higher on subtests of the Wechsler Intelligence Scale for Children which they had assessed positively than on those which they had negatively assessed. This superiority on liked over disliked subtests was also predicted to be greater for Black than for White subjects and for lower class, compared with middle class, subjects. Subjects performed significantly better on their liked than on their disliked subtests, p < .01, supporting the telic human image advanced by logical learning theory. No racial or social class differences in this effect were seen. PMID- 16367141 TI - The effects of mood upon imaginal thought. AB - The effects of mood upon imaginal thought were explored with a highly trained undergraduate female hypnotic subject. She was hypnotically programmed to experience free-floating anxiety or pleasure in varying degrees just before the exposure of combinations of three Blacky Pictures, and to produce dreamlike imagery in response to the Blacky stimuli while under sway of the mood. Data from 98 dream trials, separated by amnesia, indicated that the affective states clearly influenced imaginal processes. Blind ratings by a psychoanalyst showed the anxiety moods to be more closely associated with primary-process features characteristic of nocturnal dreams, whereas pleasure had a relatively higher incidence of daydreamlike ratings. Empirical analysis of themes yielded significant relationships of anxiety to physical injury to the self and verbal aggression toward others; pleasure was associated with circular movements and overt sex themes. PMID- 16367155 TI - The Hand Test: fifteen years later. AB - The Hand Test, a projective device developed about 15 years ago, has gained rapidly in popularity among clinical and school psychologists. A variety of reliability and validity studies have been conducted with the instrument across a broad spectrum of clients and settings, A considerable amount of developmental work has been conducted on the Hand Test since its inception. Research on this test was reviewed and discussed with the conclusion that the Hand Test has merit in many clinical applications for children, adolescents and adults. The instrument has potential in diverse assessment situations and is used today by a wide spectrum of practitioners. Research points toward the Hand Test as being a valuable quantitative multidimensional instrument which predicts behaviors. PMID- 16367156 TI - The validity of MMPI interpretations based on the Minimult and the FAM. AB - In this study 29 clinicians provided Q-sort descriptions for 36 patients' MMPI profiles prepared in the standard way or prepared according to Kincannon's (1968) Minimult or Faschingbauer's (1974) FAM. For each patient, correlations between Q sorts based on the standard profile and each short form profile were computed as a measure of interpretive similarity between the long form and short form. Correlations between Q sorts by independent raters both interpreting the long form profile served as the standard for comparison. Data analyses revealed that at least for the FAM, interpretive similarity between the short form and long form MMPI may be sufficient to warrant its clinical use for individual profile interpretation. PMID- 16367157 TI - MMPI scale development methodology. AB - A multitude of special MMPI scales have been developed without sufficient consideration for the methodology of empirical scale construction. Suggestions are presented in this article regarding the selection of items for a new MMPI scale and the tasks needed for the evaluation of a new scale. An investigator who develops a new MMPI scale should consider (a) the need for K corrections of scale scores, and (b) the possibility that the information provided by the special scale can also be obtained from the standard MMPI scales. PMID- 16367158 TI - Cross-cultural second order factor structures of the 16PF. AB - Cross-cultural personality research has generated a great amount of data on individual difference patterns in diverse cultures. One of the major instruments used in this research has been Cattell's 16PF. A major question in this research is whether the underlying personality structure is equivalent for different cultures. The present study evaluated the second order factor structure of the 16PF in 101 subjects of European ancestry and 117 subjects of Japanese ancestry. The factor structure for the Japanese was significantly different from that of the caucasian group. The caucasian results did not differ from those reported by Cattell and his associates. The implications of these results for personality theory and for cross-cultural evaluation were briefly discussed. PMID- 16367159 TI - The development and evaluation of self-esteem measuring instruments. AB - It is argued that a major failing with most existing measures of self-esteem is that they do not even attempt to take into account the value system of the individual subject. A relatively straightforward method of measuring self-esteem was proposed based on an aggregate of a subject's self-ratings in different areas of his life weighted by the relative importance of those life areas to the subject. This approach is used to develop a measure of self-esteem for Australian tertiary students. A self-esteem questionnaire was also developed, based on life areas important to other Australian college students. An evaluation of these two instruments is then reported based on 235 Psychology students at Melbourne University. Some implications of this approach to the structure of self-esteem itself are considered. PMID- 16367174 TI - Handwriting as a correlate of extraversion. AB - The validity of graphologists' claims that handwriting is a measure of extraversion was examined in this study. Three handwriting measures, line slope, letter slant, and letter width were chosen, and the relationship between these measures and extraversion as measured by the Eysenck Personality Inventory was studied. The subjects were 58 tertiary students. No significant correlations between the handwriting measures and extraversion were found, nor were there significant intercorrelations between the three handwriting measures. Thus the results did not support the claim that the three handwriting measures were valid indices of extraversion. PMID- 16367175 TI - The relationship between sexual identification and the use of defense mechanisms. AB - A distinction is made between two levels of sexual identification. The deeper level was measured by May's Deprivation/Enhancement fantasy pattern, while the surface level was assessed using the Masculinity-Femininity of the Strong Vocational Interest Blank. These measures, while clearly differentiating between college males and females, were found to be unrelated to each other. Moreover, the deeper level was found to be related to the use of "masculine" and "feminine" defense mechanisms. PMID- 16367189 TI - Construct validity of the Rosenzweig Picture-Frustration Study. AB - Projective and semiprojective techniques are both tools and tests. A schema for assessing the validity of projective devices is proposed: (1) Primary, consisting of (a) construct validity i.e., the soundness of the techniques vis-a-vis its underlying concepts; and (b) criterion-related validity, i.e., the concordance between tool results and specified external criteria. (2) Secondary, viz., pragmatic validity, i.e., the usefulness of the instrument in applied fields. Discussion then focuses on the construct validity of the conceptually-based, semiprojective Rosenzweig P-F Study. specifically the evidence on (a) the self consistency of its conceptual dimensions; (b) the sequential. social development of the Types and Directions of Aggression; (c) levels of behavior elicited; (d) sensitivity to scorable changes after induced frustration: and (e) physiological correlates oft he aggression categories. In each of these areas evidence exists in support of the validity of the P-F Study. PMID- 16367190 TI - On the validity of graphoanalysis: a rejoinder to Crumbaugh's Reply. AB - Crumbaugh (1977) reexamines an earlier study showing no validity for graphoanalysis (Vestewig, Santee, & Moss, 1976), and from considerations of the methodology and from reanalysis of the data findings some validity for the system. This rejoinder points out conceptual and statistical errors in his reply, and reaffirms the conclusion of the earlier study. PMID- 16367191 TI - The validity of locus of control dimensions for Chicano populations. AB - The multidimensional locus of control literature supported the tenability of five factorial dimensions: a) luck/fate, b) leadership/success, c) academics, d) politics, and e) respect. Contending that the contradictory locus of control findings involving Chicano populations may be due to methodological inadequacies, the purpose of the present study was to empirically determine the appropriateness of the five categories for comparing Chicano and Anglo populations. This was done by factor analyzing the responses of 203 Anglo and 244 Chicano undergraduates to Rotter's (1966) 1-E scale separately, and then comparing the corresponding factor pairs by using Cliff's (1966) congruence procedure. The luck/fate and leadership/success factors show substantial invariance across the two samples, whereas the cultural equivalence of the remaining three factors is somewhat questionable. The findings are discussed in relation to current knowledge of cross-cultural differences between Anglo and Chicano populations. PMID- 16367202 TI - An assessment of the Myers-Briggs Type indicator. AB - A comprehensive assessment of the Myers-Briggs Type indicator is presented, including a description of the four personality scales measured by the instrument, a summary of the scoring process, and an extensive review of intercorrelation studies, reliability studies, and validity studies conducted with the Indicator. Results of the studies indicate that the Myers-Briggs Type Indicator is an adequately reliable self-report inventory. The Extraversion Introversion Sensation-Intuition, and Thinking-Feeling scales appear to be relatively independent of each other, measuring dimensions of personality which seem to be quite similar to those postulated by Carl Jung. PMID- 16367203 TI - The influence of defensive styles upon the predictive validity of the Thematic Apperception Test. AB - This study investigated the extent to which defensive styles of individuals influence the predictive validity of their responses to a projective test. College subjects (n = 34) were exposed to laboratory procedures which provided the basis for identifying preferred style of dealing with aversive self-relevant information. Those who demonstrated the tendency to deny negative information about themselves and attribute it to others (projectors) gave responses to selected Thematic Apperception Test cards which were positively correlated with dependency and achievement criteria. In contrast, those who tended to deny negative information by selective forgetting (repressors) provided projective test responses which were negatively correlated with the two behavioral criteria. Although statistical significance was not consistently achieved, the trend of results supported the validity of Thematic Apperception Test performance of projectors or nonrepressors and inverse prediction from the responses of repressors or nonprojectors. PMID- 16367204 TI - Ego development and conformity: a construct validity study of the Washington University Sentence Completion Test. AB - Loevinger's conception of ego development predicts a curvilinear relation between it and conformity. Using 107 private-school boys in grades 8, 9, and 11, self report, peer-nomination, and experimental (Willis) measures of conformity were given. An archival measure, number of demerits, was also obtained. As predicted, the self-report and archival measures show maximum conformity between the Self Protective and Conscientious Stages as measured by the sentence completion test. These results are not artifacts of age or IQ. The Willis test does not correlates significantly any other variable, and the peer ratings were methodologically suspect. Thus one aspect of the conception of ego development is partially confirmed. Conformity. as measured by these typical tests, is shown not to be a unitary variable. PMID- 16367205 TI - Construction and cross-validation of an MMPI Black-White scale. AB - A robust set of MMPI items, sensitive to the effect of race (Blacks versus Whites), was constructed by compiling the results of six prior studies. Thirty two items comprised the MMPI B-W scale. The scale was cross-validated on psychiatric patients and police cadets. Statistically significant mean differences were found between races in both groups, with 83% and 60% correct racial classification, respectively. These findings suggested that when used as a clinical instrument, the MMPI might pick up nonpathology-related racial variance contributing to scale devotion biased against the black respondent, but the degree of nonpathology-related racial scale score covariance has yet to be determined. For future research, the B-W scale was proposed as a set of standard queries in studies assessing connotative meaning for Black-White contrast groups, and to index "functional" as opposed to "objective" race membership. PMID- 16367206 TI - The world hypotheses scale: rationale, reliability and validity. AB - Theoretical descriptions are presented for four different sets cognitive assumptions concerning the fundamental meaning of events. These cognitive assumptions or world hypotheses are called formism, mechanism, organicism, and contextualism. The development of a 12-item scale to measure individual preferences for these world hypotheses is described. Finally, several empirical studies are reported in which diverse behavioral correlates have been found for individuals preferences among world hypotheses. Specifically, significant results have been obtained for choice of careers, stability of friendships, and success in both group and individual therapies. PMID- 16367215 TI - The Felt Figure Replacement Technique as a personality assessment device: validity reconsidered. AB - Conclusions from an earlier study testing the validity of the Felt Figure Replacement Technique (FFRT) were questioned and three new measures of FFRT performance were developed. The reliability of these three measures, and the conventional measure of distance error, was checked using a sample of 54 female and 49 male undergraduates at Oregon State University. These subjects also completed an established personality inventory. Three of the four FFRT measures tested proved to be reliable, and it was found that models combining the three reliable FFRT measures could yield successful predictions on the personality dimensions for females but not for males; although males and females perform similarly on the FFRT. Both groups made the same kinds of errors relative to the standard placement. Also, males and females differ on only one of the three reliable FFRT measures. It was concluded that the FFRT may yet prove a valuable assessment approach, and the implications of these conclusions for personality assessment were discussed. PMID- 16367216 TI - A content validity study of the defense mechanism inventory. AB - The content validity of the Defense Mechanism Inventory was tested by 20 raters who evaluated each item in terms of which 15 defenses and three ego threats it represented. Items purportedly measuring the global defense categories of principalization, turning against self, and reversal, achieved relatively high rater agreement (over 70%) while projection and turning against Object fared poorly (29% and 39% respectively). Differential content validity was found in the levels and areas of the Inventory, indicating that the context in which items appear affect their representativeness of defensive behaviors. The individual defense mechanisms were disproportionately represented by the Inventory. Ratings suggested that aggressiveness was the major ego threat being measured by the items. Most of the problems appear correctable through rewriting many of the items. PMID- 16367217 TI - Differential effectiveness of two classification procedures on the Bem Sex Role Inventory. AB - Two procedures-a simple median split and a difference/median split method-were used to classify 102 male and 129 female college students into masculine, feminine, androgynous, and undifferentiated sex role orientations on the basis of their Bern Sex Role Inventory scores. Differences in classification occurred among nearly one-third of the subjects, and as expected, the difference/median split procedure was more successful in predicting sex-role ideology. Using this procedure, masculine-typed females were found to be most profeminist in their attitudes, while low masculinity females were most conservative. No significant differences were obtained for the male groups using either procedure. Future research with the BSRI and related sex-role measures should employ similar comparisons to determine the relative validity of the two procedures. PMID- 16367218 TI - Validation of three measures of fear of success. AB - Three new instruments designed to measure fear of success are the Horner, Tresemer, Berens, and Watson empirically derived fantasy-based scoring system, the Pappo Fear of Success Questionnaire (FOS) and the Zuckerman and Allison Fear of Success Scale (FOSS). In the absence of evidence that scores on these instruments are positively correlated or that the instruments do not measure fear of failure, correlations were calculated between these three instruments and between these instruments and the Alpert-Haber Debilitating Anxiety Scale (DAS). It was found that only the FOS and FOSS fear of success instruments were correlated positively and significantly. However, all three fear of success instruments were correlated positively at a significant level with the DAS. PMID- 16367219 TI - The Zax Information Profile: a measure of college adjustment. AB - Evaluated the effectiveness of the Zax Information Profile (ZIP), a personality test based on general fund of information, in discriminating between college students who sought mental health services and those who did not, Three successive classes of freshmen students at the Eastman School of Music of the University of Rochester were administered the ZIP. Help-seeking students were matched with non-help-seeking students for sex, age, and intellectual ability. The hypothesis that ZIP scores would differentiate between these two groups was supported by the data. The potential use of the ZIP as a predictor of college adjustment and as a useful entrance screening device was discussed. PMID- 16367229 TI - Piagetian cognitive development and primary process thinking in children. AB - On the hypothesis that, as children develop from the preoperational to the concrete level of operations, there would be a corresponding increase in the effective control of primary process ideation, 37 white, middle-class children, from 5 to 8 years old, were individually tested on eight Piagetian tasks and the Rorschach test. Piagetian Development was significantly correlated with Holt's Primary Process Manual measures of (a) Form Level, (b) Defense Effectiveness, (c) Adaptive Regression, (d) Primary Process Level 2, (e) Content Level 2, (f) Control/Defense Plus, and (g) Control/Defense Total. An analysis of variance demonstrated significant primary process differences between preoperational, transitional and concrete operational groups. Correlations between the eight Piaget tasks and the constructed mean Piaget score ranged from .65 to .91. The negative findings of other researchers, which contrast with the positive results in this study, may be explained by the narrower age ranges of their samples. PMID- 16367230 TI - Fingertip aura and interpersonal attraction. AB - Concluding from our survey of the literature that fingertip auras (Kirlian effect) might be associated with interpersonal attraction, four hypotheses were advanced to test this assertion. It was hypothesized that individuals would respond with bigger auras to (1) opposite-sex photographers as compared to same sex photographers, (2) to seductive opposite-sex photographers as opposed to normally behaving opposite-sex photographers, (3) to opposite-sex unknown peers as opposed to same-sex unknown peers, and (4) to liked as opposed to disliked same-sex persons. All hypotheses except (2) were supported. The second hypothesis was significant in a direction contrary to hypothesis. Fingertip auras are seen as a promising measurement device in the study of interpersonal attraction. PMID- 16367231 TI - First-order factor structure of the Personality Research Form. AB - To establish the first-order factor structure of the Personality Research Form, a high school sample of 327 boys and a college sample of 215 men and women were administered Form AA. Principal component analyses were made, in both samples, of the 20 scales subdivided into two subsets. Nine factors were found to be equivalent in the college and high school data. Comparison with Edwards and Abbott analyses revealed substantial agreement. It was concluded that 12 scales could be used to measure the constructs embodied in the Personality Research Form. PMID- 16367232 TI - A short measure of Eriksonian ego identity. AB - Developed a new measure of Erikson's (1950, 1959) concept of ego identity which would meet the following criteria: (a) that it be short, with an acceptable level of reliability; (b) that it be objective and easily scored; and (c) that it be free of response set contamination. A 12-item scale (Ego Identity Scale) with a forced choice format was constructed. Two further studies were conducted to test the validity of the scale. These studies showed that the Ego Identity Scale correlated as predicted with measures Of locus of control, intimacy, dogmatism, Tomkins' Left, occupational commitment, and political commitment. It also correlated in the predicted direction with measures of interpersonal trust and moral commitment though these correlations did not reach statistical significance. PMID- 16367233 TI - The common import component scoring of the Edwards Personal Preference Schedule. AB - Development of a new method of scoring the Edwards Personal Preference Schedule to yield component scores for each variable. Applications to clinical practice, research, and teaching are described. PMID- 16367234 TI - Discriminant and convergent validity of the SCL-90 in psychiatric inpatients. AB - Examined the convergent and discriminant validity of the SCL-90 in a group of 113 psychiatric inpatients and determined the degree of reactivity to several common response sets. The nine SCL-90 dimensions were found to correlate with analogous measures from other tests, and thus showed convergent validity, but were also found to correlate with nonanalogous measures, an indication of low discriminant validity. In addition, the dimensions correlated with the three MMPI validity scales, suggesting their reactivity to response bias. These findings were interpreted as indicating that the SCL-90 has limited use with psychiatric inpatients at present, although it may be useful as a brief screening device for disposition and referral. PMID- 16367246 TI - A measure of individual modernity. AB - Individual modernity may be defined as a syndrome of attitudes and beliefs, including progressivism, secularity, optimism, future-oriented perspectives, and a sense of personal efficacy. Measures of individual modernity have frequently been used in sociological and cross-cultural inquiry, but less attention has been paid to these devices by psychologists. An attempt was therefore initiated to construct- a new, brief index of modernity suitable for group administration and as free as possible from nuisance relationships to generalized distrust and anomie. An 8-item scale appearing to possess the desired attributes was developed on samples of 80 males and 139 females. Tentative norms were derived from a new sample of 766 subjects, and preliminary interpretational information was presented. PMID- 16367247 TI - Comparing the diagnostic validity of an abbreviated and standard MMPI. AB - The diagnostic efficacy of the MMPI-168 and standard MMPI was compared with a sample of psychiatric inpatients. Using traditional psychiatric measures and projective testing as a basis for the criterion diagnoses, no significant differences were found between the two MMPI forms. Rationale for the methodology employed was discussed. PMID- 16367248 TI - Tolerance for ambiguity and self-actualization. AB - This study was designed to test the hypothesis, implicit in several areas of previous research, that high tolerance for ambiguity is a cognitive style which reflects high levels of underlying psychological health. Using Self-Actualization scores on the Tennessee Self Concept Scale, 36 adult subjects were defined as relatively high or low in self-actualization with equal numbers of males and females in each group. All subjects then performed individually on the Rorschach test and the Rorschach protocols were rated reliably for degree of tolerance for ambiguity. It was found that regardless of sex the high self-actualization group exhibited a significantly higher level of tolerance for ambiguity than the low self-actualization group. The finding was interpreted as confirmation of the study's hypothesis. PMID- 16367258 TI - Primary process thinking in the same children at two developmental levels. AB - Analysis of primary process thinking of the same children at kindergarten level and at grade four revealed that amount of primary process thinking was relatively constant with development, contrary to predictions based on psychoanalytic theory. However, the two facets of primary process thinking, i.e., drive related ideation and formal deviations of thought follow different lines of development with the former increasing and the latter decreasing in quality and type with age. PMID- 16367259 TI - Validating a measure of children's others-concept through population and behavior variables. AB - Data are presented to support the Paired Hands Test (PHT) as a valid measure of the others-concept in children. This construct is defined as a person's general expectancies or perceptions about other people along a positive-negative continuum. Behavioral data are reported which replicate findings from earlier, smaller studies that children who have relatively high others-concepts are likely to interact more positively in small, unsupervised groups than children having relatively low others-concepts. Population data are also given, based upon 1,235 PHT test scores broken down by such variables as sex and socioeconomic status. Additionally, content validity inherent in the nature of the test is discussed. The conclusion drawn is that the data can be best explained by employing the others-concept as a theoretical construct measured by the PHT. PMID- 16367260 TI - Refining computerized test interpretations: an in-depth approach. AB - Comprehensive individual psychological evaluations of a small sample of subjects were used to identify shortcomings and suggest improvements in a computerized system for interpreting 16PF and CAQ scores. This approach is offered as a supplement, rather than an alternative, to the usual large sample correlational method. It is argued that such procedures mask desirable refinements by substituting a large number of subjects for detailed and accurate assessment of individual cases. Results of this analysis supported the general accuracy of the computerized system, but revealed gaps in the information provided and suggested alternatives for perfecting the system. PMID- 16367261 TI - Testwiseness: some evidence for the effect of personality testing on subsequent test results. AB - The assumption is presented of the test-taker as a hypothesis-generating organism who can become "testwise." Testwiseness is defined as a stable skill, acquired by test-taking experiences, by which an individual can make test responses conform to a desired response pattern. Forty-three college students completed two forms of The Personality Research Form (PRF) and a rank ordering of their predicted personality need pattern. Results show significantly higher correlations of PRF predictions in the second administration. Analyses show PRF profiles, not predictions, to have been modified. Furthermore, high testwise subjects had higher needs for Understanding and Nurturance, and lower needs for Aggression and Defendence than low testwise persons. The importance of considering testwiseness, given trends in society encouraging access to psychological records, is discussed. PMID- 16367271 TI - Order effects in the administration of personality measures: the case of the Self Consciousness Scale. AB - The pitfalls of serial administration of personality measures as a research selection device were explored. Three samples of subjects were administered the Self-Consciousness Scale (Fenigstein, Scheier, & Buss, 1975) either first or last in a series of questionnaires. As predicted, those who responded to the measure last obtained higher private self-consciousness scores. However, the strength of the effect was moderated by the number of other measures included in the testing. Unexpectedly, public self-consciousness scores also were influenced by order of administration of the measure. Subjects who were administered the scale last evidenced lower public self-consciousness scores although this effect reached significance in only one of three samples of subjects. The broader implications of these findings for researchers who employ mass test administration procedures to select subjects for personality research are considered. PMID- 16367272 TI - Construction and validation of a measure of authoritarian personality. AB - This paper reports the results of a series of studies in which the construction and validation of a behavioral measure of authoritarianism is reported. Factor analysis of already established behavioral scales (e,g., Ray's (1976) Directiveness scale; the Buss-Durkee (1957) Hostility inventory, etc.) and items especially written for the study resulted in a Revised F scale with a reliability (Cronbach's alpha) of .79 and a rpN of -.45. The items loaded significantly on four factors, namely, Dominance/Leadership, Achievement motivation. Interpersonal conflict and Verbal hostility. Furthermore, validity studies revealed the scale to possess satisfactory validity. It is suggested that this measure be seriously considered as an alternative indicator of levels of authoritarianism. PMID- 16367283 TI - Distinguished Contribution Award: the phantasy behind the face. PMID- 16367274 TI - Researchers, sex differences, and the Holtzman Inkblot Technique: a reply to Rice, Greenfield, Alexander, and Sternbach. AB - Two aspects of a previous article by Rice, Greenfield, Alexander, and Sternbach (1976)--their errors with regard to a previous article by Thorpe and Swartz, and their statement regarding sex differences on Holtzman Inkblot Technique variables -are described and discussed. PMID- 16367284 TI - Ruminations on teaching projective assessment: an ideology, specific usages, teaching practices. AB - This paper was written in reaction to current dissatisfaction with projective techniques. A credo for use of projective techniques emphasizes accountability, phenomenological equality, and an eventual "giving-away" of assessment procedures through education. The primary legitimate usage is nonpsychometric; an interview focused on growth in various contexts. Secondary usages include examples of procedural or methodological effects to provide specific information. Training practices germane to the credo and legitimate usages are presented. PMID- 16367285 TI - The elusive homosexual: a reply to Stone and Schneider. AB - In a rebuttal of the Stone and Schneider validation study of the Wheeler signs of homosexuality in the Rorschach, it was argued that (a) homosexuality is not a proper clinical diagnosis, (b) homosexuals do not differ in psychological functioning from hetereosexuals, and (c) the Wheeler signs represent an atomistic approach of use only to those defensive clinicians unwilling to encounter the unique character of their clients. PMID- 16367286 TI - Exposure duration as a confounding methodological factor in projective testing. AB - Exposure duration has been found to have a strong impact on the affective value attributed to a variety of stimuli. The purpose of the study reported here was to examine the impact of this variable in projective testing, and im particular in the affective value subjects attribute to the TAT cards. Using 34 undergraduate students, a linear decrease in pleasantness was found as a function of exposure duration. Factors leading to variation in exposure duration might therefore be confounded with the interpretations made of a subject's protocol. It was suggested that the time a subject views a projective test stimulus should be controlled in the administration. PMID- 16367287 TI - An empirical comparison of Maslow's and Murray's needs systems. AB - The Work Motivation Inventory (WMI), a measure of Maslow's hierarchy of needs, and the Edwards Personal Preference Schedule (EPPS), a measure of Murray's manifest needs, were administered to 372 undergraduates. The two instruments were compared using canonical analysis. The analysis revealed three significant relationships between components of the two instruments. The first relationship supported Maslow's need hierarchy in general and its measurement by the WMI. The second suggested a fluctuating relationship between giving and receiving help and the levels of Maslow's hierarchy. The third relationship suggested that need for Achievement is associated with the intermediate levels of Maslow's hierarchy. PMID- 16367288 TI - Value changes in third generation Japanese Americans. AB - To compare the acculturation of third generation Japanese Americans with a contemporary Caucasian college student population 275 Japanese Americans (106 males, 169 females), and 346 Caucasian American college students (136 males, 210 females), were given the Edwards Personal Preference Schedule, which describes personality in terms of 15 needs. The results indicated that while acculturation has taken place among the Japanese Americans, when their scores are compared with a contemporary Caucasian college sample, the Japanese Americans still express a significantly higher need for order, succorance, and abasement, and a significantly lower need for change and heterosexuality. PMID- 16367289 TI - Measurement of ambiguity tolerance. AB - Presented definitions for the construct of ambiguity tolerance. The measure of ambiguity tolerance (MAT-50) had high internal reliability (r = .88) and high test-retest reliability (r = .86) over a 10-to-12 week period. A content analysis of the measure and a subjective analysis by 20 graduate students indicated adequate content validity. A multivariate comparison with two other ambiguity measures, two rigidity measures, and a short dogmatism measure provided strong evidence for criteria-related validity. Finally, four independent empirical studies showed good construct validity. PMID- 16367290 TI - Diagnostic sex-role bias: how can we measure it? AB - A new instrument was designed to measure sex-role bias towards children. The instrument, the Chasen Diagnostic Sex-Role Bias Scale, is composed of case histories focusing on the "somewhat unhealthy" aspects of two main factors of behavior: activity (composed of independence and aggression); and passivity (composed of dependence and nonaggression). Validity and reliability coefficients were reported as well as a formula by which to use the scale to diagnose individual bias, in addition to group bias. In using the scale on a national sample of school psychologists, it was determined that differences in Diagnostic Sex-Role Bias scores exist, as do wide variations about what is considered healthy and unhealthy behavior among girls and boys. PMID- 16367291 TI - The Activity Preference Questionnaire: is the forced choice format necessary? AB - While the Activity Preference Questionnaire (APQ) appears to provide unusually pure and subtle measures of anxiety proneness, its forced choice format involves several drawbacks. Data are presented which suggest that a rating format can be used which avoids the disadvantages of the forced choice format but incorporates its advantages. PMID- 16367302 TI - Change in college students' attitude toward child-bearing from 1971 to 1973. AB - Two samples of college students, one obtained in 1971 and one in 1973, were asked to draw a picture of a happily married couple. Some subjects were asked to draw the couple after two years of marriage and some after five years of marriage. The pictures were scored for the presence or absence of children as an estimate of the attitude that children are necessary for a happy marriage. A large decrease in the "presence of children" from the 1971 to 1973 sample was interpreted as indicating that fewer college students now accept the notion that children are necessary for marital happiness. PMID- 16367303 TI - Parents' predictions of their children's perceptions of family relations. AB - Thirty children from 15 families were administered the Bene-Anthony Family Relations Test. Their parents then assigned test responses as they imagined their children had. With only two exceptions, both involving first born boys, parents predicted their children's responses beyond chance expectation. However, significant discrepancies were found between children's responses and their parents' predictions. Parents tended to overestimate their own centrality in the family perceptions of their children, particularly perceptions involving negative feelings, and to underestimate children's negative feelings involving their siblings. PMID- 16367304 TI - A test of the Zucherman heterosexual scales. AB - Female and male undergraduate students enrolled in the introductory psychology course were administered the Zuckerman heterosexual scale as part of a larger battery of tests. The ordering of items for males was very similar to Zuckerman's with two minor exceptions. Female subjects, on the other hand, reported more sexual experiences of all kinds than did Zuckerman's sample, and two major differences in ordering Were noted. The coefficients of reproducibility for males and females were .928 and .954 respectively which were almost identical to those of Zuckerman. The rank-order correlation between the ranking of the items in the male and female scales was .958. PMID- 16367305 TI - A note on the PMT as an objective test of achievement motivation. AB - Studied the properties of the PMT when used with 102 male cadets of the Australian Army's Officer Training Unit. Internal consistency of the PMT was high, while intercorrelations of the test with other measures in the battery supported the validity of the PMT as a measure of achievement motivation, but indicated its limitation in a practical situation of officer selection. The English version of the PMT would benefit from a rewriting of Items 81 and 88. PMID- 16367319 TI - Solomon design analysis of multiple-choice Rorschach animal content. AB - The Solomon four-group design was used to study the effects of a persuasive message on a selected multiple-choice Rorschach index--animal content. The independent variable elicited behavior in a predictable manner. Pretesting as a main effect was not significant, but as an interactional effect obviated the effect of the persuasive message. Although knowledge of test rationale can elicit behavior that conforms to experimental demand characteristics, some subjects nonetheless acted in defiance of such information. A condition for defiance in this experimental arrangement, however, was pretesting. Other possibilities regarding the study of compliance behavior and the use of pathognomonic indicators were suggested. Ethical issues were raised. PMID- 16367320 TI - Sources of male chauvinism in the TAT. AB - Potential sources of antifemale bias in TAT stimuli were evaluated by having 358 undergraduate subjects rate 17 male and 17 female TAT figures on 7-point anchored scales. Data from the five independent rating conditions were examined by 2 x 2 ANOVA. Biases toward greater Mental Health and Intelligence for female figures were seen to be insufficient counterbalancers of biases toward greater Cultural Favorability and Identification for male figures. Achievement status was rated equivalently. TAT stimuli appeared to show a "built in" source of male chauvinism systematically "pulling" male-sex identification. Potential for unfavorable clinical evaluation was seen to be greater for female TAT subjects compared with male subjects. PMID- 16367321 TI - Need associations of male prisoners: a multi-dimensional assessment of Murray needs. AB - Previous studies have revealed quantitative differences between delinquents and nondelinquents in the strength of Murray needs. To provide clarification and elaboration of such past findings, need associations derived from the Picture identification Test were analyzed by t test and discriminant analysis for a group of 112 male prisoners and 112 male nonincracerated normals matched with the prisoners for age and education. Greater than chance (.05 level) numbers of association score differences were obtained between the two groups. Differences in need association scores were interpreted in light of past findings. Need association differences were revealed for the Achievement, Affiliation, Sex, and Order needs among others. PMID- 16367322 TI - An investigation of the Mini-Mult validity scales. AB - Examined the ability of the Mini-Mult validity scales to detect invalid MMPI profiles. When 34 invalid MMPI profiles were rescored with the Mini-Mult only 17 of the 34 profiles invalidated by the full MMPI were detected with the Mini-Mult. This included 14 of 27 profiles invalidated by an elevated F scale; 2 of 4 profiles invalidated by an elevated L scale and 1 of 3 profiles invalidated by an elevated K scale. Only 14 of 27 profiles invalidated by an F-K ratio of K11 were detected. When new conversion values for the Mini-Mult were utilized, the detection rate improved considerably for the F scale and the F-K ratio. PMID- 16367323 TI - Relations between embedded figures test performance and dimensions of the I-E scale. AB - Relations between an embedded figures measure of field independence and various dimensions of Rotter's internal-external locus of control scale were investigated. The general absence of significant correlations suggests that these variables are essentially unrelated. PMID- 16367324 TI - Validity of the Internal-External Scale in its relationship with political position. AB - The study was designed to interpret previous reports of a relationship between left wing political position and externality on Rotter's (1966) Internal-External (I-E) Scale. I-E items phrased in the first person were associated with a criterion measure of locus of control, and externality on the scale predicted inferior performance in a skill-controlled task. However, political position was related to neither the criterion measure of locus of control nor to successful performance in a skill-controlled task. Thus, there was no evidence that political position was related to locus of control in spite of its relationship with the I-E Scale. There was discussion of implications for the validity of the I-E Scale and for studying relationships between personality and political variables. PMID- 16367329 TI - Further validation of a measure of individual modernity. AB - SUMMARY: Individual modernity may be defined as a cluster of beliefs and attitudes including progressivism, autonomy, secularism, and a sense of personal efficacy. An 8-item scale to assess this constellation was previously published. The present analysis reports new validational data. Factor analysis showed the scale to be significantly loaded (.74) on a modernity dimension, with minimal loadings (-.14 and -.16) on factors for alienationanomie and authoritarianism. For 60 assessees modernity correlated .35 with criterion u ratings by observers. Interviewers' Q sort and adjectival descriptions of 360 subjects stressed independence and breadth of interests for high-scorers versus conventionality and narrowness of interests for low-scorers. These descriptions also suggested that women ranking high on modernity will be less favorably perceived than men of comparable standing. PMID- 16367340 TI - Further investigation of Coleman's Level Hypothesis. AB - Volunteers (16 male and 17 female) from introductory psychology courses were used to investigate Coleman's rationale in support of the Levels Hypothesis. Projective stimuli from parts of the Rorschach, TAT, and Sentence Completion techniques were presented, and subjects asked to rate how much the stimuli and their responses said something about themselves, their fathers, or mothers, to determine their awareness of the personal relevance of the stimuli and responses. Results indicated that both the stimulus and response ratings varied as expected at the .01 level of significance, thus supporting Coleman's rationale. PMID- 16367341 TI - Some structural considerations in the relationship between M and empathy. AB - After reviewing some of the difficulty around defining empathy, this paper draws attention to three underemphasized structural factors in the empathic tie between self and other. The following distinctions are drawn: (a) empathy vs. fusion with the object: (b) empathy vs. a narcissistic attachment to the object: and (c) empathy vs. an intuitive but fragmented orientation to part-objects. The relationship between Rorschach's M and empathy is reexamined in light of these considerations, and it is concluded that the experience of kinaesthesia on the Rorschach reflects some, but not all, of the components of empathy. PMID- 16367342 TI - Validity and student acceptance of a graphoanalytic approach to personality. AB - Forty-eight handwriting specimens were rated by 6 handwriting analysts on 16 personality variables. Correlations of analysts ratings and personality inventory scores were nonsignificant. Analysis of variance showed that there were significant differences between the analysts on their judgments of 10 of the 15 traits. Subjects were also given the reports prepared for them and a bogus report and asked to rate the analysts' assessments for accuracy. Subjects were able to choose 2 of the 15 traits with significant accuracy on the real report when considering individual traits. There were no significant differences between the accuracy ratings on the real and bogus reports on overall rating. It was concluded that the analysts could not accurately predict personality from handwriting. PMID- 16367343 TI - Susceptibility to faking of a sentence completion test of ego development. AB - Effects of faking on a sentence completion test. of ego development, explored in five experiments, partially confirmed the developmental hypothesis that persons can lower their ego level test scores but not increase them. When faking high on retest, subjects scores either stayed the same or increased by about a half step. Only intensive study of ego development seemed genuinely to increase scores. When faking a bad impression or a low ego level, subjects' scores decreased, typically to the pre-Conformist level. Unexpectedly, attempts to make a good impression lowered high scores, especially for women, Because the test is susceptible to factors which can artificially lower scores, careful planning and evaluation of testing conditions must accompany test use. PMID- 16367344 TI - Authoritarian personalities, 1950-1973. AB - This study concerns a comparison of American college graduates of the post Kent State era with those of the post Hitler era as to authoritarian type personality. A short 17-item scale for authoritarian personality traits was devised and found to correlate with the F Scale. This was administered to 314 college graduates in 1950. It was again administered to 400 graduates of the same college in 1973. The results show that the 1973 group is less authoritarian than the 1950 group. PMID- 16367345 TI - The dimensions of nonauthoritarianism. AB - The converse of authoritarianism has not been conceptualized very clearly though scales purporting to measure nonauthoritarianism are among the most commonly used in. personality inventories. Using the Autonomy scale of the Omnibus Personality Inventory (a derivative of the original California F scale), as administered to a sample population of beck and white college freshmen, the dimensions of nonauthoritarianism were explored. A factor analysis of the scale creates three components (anti-authoritarianism, anti-conventionalism and open-mindedness) that are conceived of as successive phases in the weakening of authoritarianism. This phase hypothesis is supported with evidence from Guttman scales and a correlational analysis of how black and lower-class as compared to white and middle-class students shift from one phase to another over the course of the freshman year. PMID- 16367361 TI - Construct validity of the Holtzman Inkblot Anxiety and Hostility scores. AB - The group version of the Holtzman Inkblot Technique (HIT), the IPAT Anxiety Scale Questionnaire (ASQ), the Manifest Hostility Scale (MHS), and the Hand Test (HT) were administered to 72 college subjects in an attempt to clarify the nature of the HIT Anxiety (Ax) and Hostility (Hs) scores. The Ax score was found to correlate significantly with the ASQ total score which represents both overt and covert anxiety. The Hs score was found to correlate significantly with the MHS, which is sensitive to moderate levels of verbal hostility, but not with the HT Acting Out Ratio (AOR), which is strictly a measure of overt aggressiveness. The HIT Ax and Hs scores were concluded to relate most closely with measures that encompass a wide spectrum of the variable in question. PMID- 16367362 TI - Quantitative difference in the experience of state-trait anxiety in emotionally disturbed and normal children. AB - In order to investigate the state-trait anxiety distinction in emotionally disturbed and normal children, the responses of 120 emotionally disturbed children and 126 normal children to the State-Trait Anxiety Inventory for Children (Spielberger, 1973) were factor analyzed separately. For the emotionally disturbed group two A-State and two A-Trait factors were found with the A-State factors accounting for significantly more of the variance than did A-Trait factors. For the normal group four A-State and two A-Trait factors were found, but there was not a significant difference between the amount of variance accounted for by A-State and A-Trait factors. Results were discussed as supporting the state-trait distinction and as having several implications for future research on anxiety. PMID- 16367370 TI - Recent research trends in testing. AB - Recent trends in test research as measured by references in Buros' Mental Measurements Yearbooks were investigated. The 24 tests with the greatest number of publications were identified, and growth trends of five major tests (Rorschach, MMPI, TAT, Stanford-Binet, and SVIB) were shown. The correlation between test publications and test usage was .25. The authors speculated that future research trends might include increasing psychometric attention to behavioral assessment and test modifications for minorities. Based on a survey of psychologists' judgments of test quality, the authors concluded that good psychometric properties do not test leaders make. PMID- 16367371 TI - A measure of needs to assess the stimulus characteristics of TAT cards. AB - The stimulus demands of TAT cards were evaluated by means of a TAT-Adjective Rating Scale. Seventeen needs were measured through adjective ratings. Mean needs were reported for 191 males and 85 females. Variability was used to develop a measure of stimulus ambiguity. Also investigated was (a) the partitioning of variance for subjects, cards, needs and their interactions, and (b) sex differences were evaluated by means of analysis of variance. PMID- 16367372 TI - The predictive efficacy of three MMPI short forms. AB - In order to investigate the predictive efficacy of three MMPI short forms, the standard form answer sheets of 50 inmate volunteers and 44 clinic outpatients were scored separately for Faschingbauer's, Hugo's and Mini-Mult scales. T-score correlations and t tests between the standard form and each short form, as well as comparisons using Lacks' criteria were reported. Results indicated that the Faschingbauer form was superior to the other two forms. Results were discussed in terms of the populations tested and in reference to the different methodologies used in developing each of the short forms. PMID- 16367373 TI - A system for assessing personal responsibility: validity, reliability and rater trainability. AB - Sixty-two subjects completed the California Psychological Inventory, the Rotter External-Internal locus of control scale and an audio-taped discussion of their personal problems. The audio-taped problems were rated on a five-point level of personal responsibility scale and were compared with the scores on the California Personality Inventory and the Internal-External locus of control scale in a correlation matrix which was subjected to a factor analysis. The results from these analyses supported the hypothesis that the Personal Responsibility Rating System has construct validity as a measure of psychological health. Study II assessed the trainability of the Personal Responsibility System. With a four-hour training program it was found that graduate students could be taught to rate personal responsibility in a reliable manner. PMID- 16367374 TI - Psychodynamic inferences from the Strong Vocational Interest Blank. AB - A systematic psychodynamic approach to personality appraisal using the Strong Vocational Interest Blank is presented. The psychoanalytic concepts of transformation of infantile impulses, ego style, and structural self representation are discussed in relation to issues of vocational choice. A method for making psychodynamic inferences from the SVIB is then put forth. To illustrate the method, "blind" personality evaluations, of three late adolescents, using only their SVIB profiles, are presented. The results of each appraisal are discussed in relation to independent information about the client and the counseling process. PMID- 16367375 TI - Subjective ratings of Cattell's 16 Personality Factors. AB - The Sixteen Personality Factors Questionnaire (16PF) was administered to 69 subjects. Each subject was asked to rate himself on the 16 personality factors on the 16PF. The 16PF scores and the self-ratings were compared by canonical analysis. Three significant relationships were identified. The relationships primarily involved indices of social interaction from the 16PF and indices of internal traits from the self-ratings. PMID- 16367376 TI - Social desirability scale values of locus of control scale items. AB - Three groups of subjects rated each item in Rotter's I-E Scale for its social desirability value. Social desirability scale values (SDSV) of paired items were compared with one another. Results indicated that paired items are not sufficiently similar in their SDSVs, thus enabling subjects to respond on the basis of social desirability. All three groups rated internal items as being significantly more socially desirable than external items. The correlations between the SDSV ratings of the three groups were highly significant. The results indicate that the scale, as it is currently used, is seriously deficient. PMID- 16367377 TI - Dimensionality of three adult, objective locus of control scales. AB - The current conceptualization of the locus of control construct as unidimensional was challenged. The Rotter I-E scale, James I-E scale, and Adult Nowicki Strickland I-E scale were examined with regard to their reported psychometric properties. The three scales were factor analyzed using a principal components method with varimax rotation. Results indicated a multifactor makeup for the three scales. It was suggested that attempts to measure a generalized expectancy for control be abandoned and that, instead, measurement attempts focus on the development of instruments specific to the behavioral and situational domains for which behavioral understanding, prediction, and control are sought. PMID- 16367378 TI - Buffers items, insight, and the Marlowe-Crowne Social Desirability Scale. AB - The influence of buffer items on response to the Marlowe-Crowne Social Desirability Scale (MC) was examined. Thirty-four college student subjects received MC questionnaires with intermixed extraversion and neuroticism items, while 35 others received separate MC and extraversion-neuroticism questionnaires. MC means under the two conditions differed nonsignificantly, and confidence interval estimation spoke against the likelihood of any appreciable buffer item effect. Little insight into the purpose of MC was exhibited--less than half that previously reported by Shrauger (1972)--nor was it related to the buffer item manipulation. PMID- 16367387 TI - Cognitive anxiety: a method of content analysis for verbal samples. AB - The work of such psychologists as Kelly, McReynolds, Epstein, and Lazarus suggested the need for a measure of cognitive anxiety and provided a definition of that construct. A method of content analysis of verbal samples was devised and found to have adequate interjudge reliability. Normative data for five groups of subjects were provided. The validity of the measure as representative of a reaction to being unable to anticipate and integrate experience meaningfully was demonstrated in (a) the higher scores of groups of subjects who were currently coping with new experiences than those who were not, (b) the significant correlation of its scores with a state rather than trait anxiety measures, (c) the variability of its scores over time as observed in a generalizeability study, and (d) the higher scores of subjects when they were dealing with experiences for which meaningful anticipation was relatively difficult. PMID- 16367388 TI - Filler items and social desirability in Rotter's locus of control scale. AB - Three experiments were conducted to determine whether the use of filler items in Rotter's I-E scale fulfills its stated objective of obscuring the purpose of the scale. Subjects were undergraduate psychology students. Experiment 1 compared the effects of filler items and sex of subject on 145 subjects I-E scores. Experiment 2 examined effects of filler items and sex on I-E responses of 210 subjects under standard and fake good instructions. Experiment 3 examined subjects' ability to discern the purpose of the I-E scale, with and without the filler items. Filler items did not affect I-E score, impede subjects from faking good, or obscure knowledge of the purpose of the I-E scale. Sex differences occurred in faking good, knowledge and certainty of correctness of knowledge. PMID- 16367389 TI - The self-consciousness scale: a discriminant validity study. AB - A recent Self-Consciousness Scale (SCS) measures Private and Public Self Consciousness and Social Anxiety. The present research was intended to establish discriminant validity for the two Self-consciousness components with respect to several other variables. 105 male undergraduates completed the SCS, the Otis Test of Mental Ability, an abbreviated Edwards Personal Preference Schedule (measuring need for achievement), a Test Anxiety Questionnaire, and the EASI Temperament Survey. Private Self-consciousness did not correlate significantly with any other measure. Public Self-consciousness correlated weakly with the temperaments of Sociability and Emotionality. The Self-consciousness components of the SCS thus appear to be relatively independent of the other measures tested. PMID- 16367390 TI - A psychometric investigation of Eysenck's personality inventory. AB - Eysenck's personality inventory (EPI) was administered to 653 females and 666 males. Item analyses were carried out for the subsamples and the combined sample. In addition hierarchical oblique factor analysis was applied to the data. The purpose of both item and factor analysis was to examine the psychometric properties of the EPI in regard to (a) the homogeneity of the scales, in terms both of internal and external item-scale relationships (b) the univocality of the scales assessed in factor analytic terms. It was found that the neuroticism scale was superior to the extraversion scale in both these respects. PMID- 16367399 TI - The medical model revisited: a humanistic reply. PMID- 16367400 TI - Influences of examiner differences on rorschach productivity in children. AB - Rorschach test protocols for a matched sample of male and female subjects in the child and adolescent age range were scored for total responses. The data were analyzed for evidence of interactions between sex of experimenter and sex and age of subject. Consistent differences in total numbers of responses elicited by different examiners were identified. The productivity of both male and female subjects was found to be significantly more variable for male experimenters than for female experimenters. Alternative explanations for the results include greater structuring of the test situation by female experimenters or greater variability of behavior by male experimenters in relation to different subjects. That these explanations can also account for the sex interactions reported by others is recognized. PMID- 16367401 TI - A group version of the Stroop Color and Word Test. AB - The uses of the Stroop Color and Word Test have been limited because of the lack of a standardized form, particularly one which could be used for both individual and group administrations as demanded by the requirements of a research project. The present version of the test counted the number of items completed by the subject in a 45-second time period. In the individual version the subject said the words out loud while in the group version he said the words to himself. Results indicated that the two versions were equivalent to each other as well as to forms requiring the subject to read a whole page of items. It was felt that a group form would greatly enhance the research capabilities of the Stroop Test. PMID- 16367402 TI - A test of the dimensionality assumptions of Rotter's Internal-External scale. AB - Examined two assumptions about the dimensionality of Rotter's I-E scale: First, the bipolarity of the two statements within each item pair; second, the unidimensionality of the overall construct. A revised I-E scale was constructed in which each of the statements from the original I-E scale was presented separately and the 367 subjects could accept or reject them independently. Correlations between statements within item pairs were low anti negative with only seven exceeding a value of -.20. The responses to the statements were factor analyzed resulting in six interpretable factors when rotated. Of the 23 item pairs, there were 11 pairs which loaded on the same factor in opposite directions while the other 12 loaded on different factors or on the same factor but in the same direction. A subsample consisting of 152 subjects also took the original I-E scale. The correlations between factor scores on the 6 factors and the original I E score varied from .15 to .52. Both assumptions regarding Rotter's I-E scale were found untenable. Instead, it is suggested that the forced choice format be abandoned and the construct be thought of as a second order factor. PMID- 16367403 TI - Psychobiographical assessment. AB - Psychobiography, the analysis of public persons by competent clinicians, is discussed as a possible assessment technique. Its re-emergence has emanated from recent attempts to psychoanalyze President Nixon, Adolph Hitler, and Emily Dickinson. Its position in relation to traditional personality assessment is considered as well as major previous efforts. Psychobiography is now a part of the curriculum at several leading universities and suggestions are made for future work. Because clinically unsophisticated writers from other disciplines are currently pursuing the trend, clinical psychologists should become involved to provide society with a more accurate psychological perspective. PMID- 16367406 TI - Relationships Between the CPI, MMPI, and locus of control in a nonacademic environment. AB - Rotter's Internal-External (I-E) locus of control scale, the MMPI, and the CPI were administered to a sample of 100 males heterogeneous in age and education level. Relationships between I-E and the other measures were examined and a substantial number of significant correlations were found. Considerable variance in I-E scores could be accounted for by similar variance in MMPI and CPI responses, but a discriminant analysis failed to enable reliable predictions of I E from other scales. The findings qualify the interpretation of correlational relationships found in previous research concerning I-E and suggest specific directions for future investigations. PMID- 16367407 TI - Adjective check list descriptions of Freud and Jung. AB - Personality characteristics of Freud and Jung were assessed by 17 members of a psychology class familiar with biographical and autobiographical material of these subjects by means of Gough's Adjective Check List, Terms used in describing the men and ACL profiles scored on composite lists were consistent with expectation, Blind interpretation of profiles by the test author accorded with knowledge of the subjects. On special scales for "intellectence" and "origence" the subjects fell high on both dimensions which is typical of creative persons. PMID- 16367414 TI - Retest reliability of the Rosenzweig Picture-Frustration Study and similar semiprojective techniques. AB - Surveys the evidence for the reliability of the Adult, Adolescent and Children's Forms of the Rosenzweig Picture-Frustration (P-F) Study. Analysis of variance and split-half methods, which assume item homogeneity and internal consistency, may be appropriate to psychometric tests, but they are not apposite to projective and semi-projective techniques. The implications of applying such criteria are critically examined, with the P-F Study as an example to demonstrate their nonapplicability. Investigations on retest (total test) reliability from the laboratory of the senior author and from those of other investigators demonstrate statistically significant consistency for the main scoring categories of the P-F, especially those relating to directions of aggression. Implications for other semiprojective techniques are expounded. PMID- 16367415 TI - Influence of instructions on expectancy effects in Rorschach testing. AB - Videotaped Rorschach instructions from a previous study in which two experimenters had biased subjects for high productivity and two experimenters had biased subjects for low productivity were played (audio or combined audio-visual channels) to matched subjects who then took the free association portion of the test. Biased results were obtained for one experimenter only. These did not interact with communication channels. It was concluded that experimenter expectancy effects in Rorschach testing are not transmitted simply by exposing subjects to biased experimenters' instructions, but instead depend upon experimenter attributes which may or may not manifest themselves during the instructional phase. Limited evidence suggesting that expectancies are transmitted as effectively by audio as by a combination of audio and visual cues was obtained. PMID- 16367428 TI - Learning and maintaining rules for decreasing judgment accuracy. AB - A meeting of Psychologists Against Tough-Hearted and Offensive Scientists (PATHOS) was convened to redress the damage done by judgment researchers who have degraded the art of clinical decision making. A special task force drafted a rebuttal and mission statement, which is presented in its entirety. PMID- 16367429 TI - A methodological note on content analysis: estimates of reliability. AB - The reliability of measurements obtained from a dictionary-based form of content analysis was investigated in this experiment by tape-recording subjects as they spoke about a predetermined topic in two separate sessions 1 week apart. Subjects were randomly assigned to speak in one of two contexts. The content of transcripts of these sessions was analyzed using adaptations of the Harvard III Psychosociological Dictionary and the General Inquirer content analysis program (Stone, Dunphy, Smith, & Ogilvie, 1966). Reliability was high over time, both within and between sessions, and there were few differences observed between contexts. Results are discussed in terms of their implications for the use of content analysis in personality research. PMID- 16367430 TI - Measures of eating-related characteristics for the general population: relationships with temperament. AB - A questionnaire that assessed a broad range of eating-related characteristics for unselected, normal subjects was factor analyzed in a two-step process proposed by Comrey (1984). Twelve "factored homogeneous item dimensions" were identified first and yielded three primary-level factors in a second factor analysis: Predisposition to Obesity (including Dieting and Preoccupation with, and Fear Of, Gaining Weight), Uncontrollable Urges to Eat (including Eating Momentum Beyond Control, Food a Panacea and Constant Temptation, and Secret Binging), and Predisposition to Anorexia (including Insufficient Eating Obvious to Others, Food Phobia, Inability to Eat, and Vomiting After Meals). The three primary-level factors were positively intercorrelated and exhibited significant positive, though weak, correlations with a measure of trait arousability. Also, weak results tentatively indicated that individuals with more pleasant and/or more arousable temperaments were less likely to be overweight. Subjects reported sharply higher levels of food consumption when feeling "depressed" (i.e., bored, lonely, sad) than when feeling "distressed" (i.e., uncomfortable, anxious, in pain). Uncontrollable Urges to Eat correlated positively and significantly with self-reports of food consumption while depressed, showing that those lacking control over eating ate especially more while feeling bored, lonely, or sad. Predisposition to Obesity correlated negatively and significantly with self reports of food consumption while distressed, showing that those tending more toward obesity ate less while upset or anxious. PMID- 16367432 TI - Where have we been and where are we going? Reflections on 50 years of personality assessment. PMID- 16367434 TI - Bruno Klopfer: a Rorschach pioneer. AB - Bruno Klopfer is perhaps best known for his pioneering work on the Rorschach Inkblot Test. His efforts in developing and popularizing projective techniques were to have a profound impact on the development of psychological personality testing. In fact, prior to the introduction of Exner's comprehensive Rorschach scoring system, the Klopfer scoring system was the most popular among practicing clinicians (Exner & Exner, 1972, p. 13). Professionals in clinical psychology and psychiatry are certainly very indebted to Bruno Klopfer. PMID- 16367435 TI - Rorschachbound: a 50-year memoir. AB - Celebrating the 50th anniversary of the Journal of Personality Assessment evokes memories of years past when I participated in its birth. Therefore, in recording what I consider significant events in Rorschach's stormy history, I am placing them within an autobiographical framework. Originally introduced into America by David Levy in the early 1920s, the Rorschach attracted students from many disciplines who revolted against the "trait" psychology that pervaded the psychological scene at the time. For 30 years, we accepted Rorschach enthusiastically as a global method that could study the human personality as a whole and in depth. In the late 1960s, we were challenged because of repeated failures in validating research. Interest declined in its development and application. Since 1970, however, it has emerged with more strength and vigor on firmer theoretical foundations and sounder methodology. Although the method is one of the most frequently required, requested, needed, recommended, and used in many clinical settings, it is still challenged today, chiefly because of overemphasis on the nomothetic approaches in research and failure to consider interactions between personality dynamics, behavior, and situational and sociocultural variables. Those of us whose lives have long been entwined with the history of the Rorschach should step back from the contemporary canvas and contemplate the future of our method, t suggest that we develop alternative conceptual models and greater diversity of research methods and modes of thought. The Journal of Personality Assessment can enhance the future of the Rorschach by giving greater prominence to clinically oriented research, notably to the idiographic and interactional aspects of the method. In so doing, the journal can help the Rorschach reach its real goal of complete scientific validity. PMID- 16367436 TI - The Lowenfeld Mosaic Test: review of its use. AB - The Lowenfeld Mosaic Test (LMT), despite rich potential for yielding information about ways in which subjects experience themselves and the world, has for many years gone unstudied and unused. A forthcoming book on this test, the first in over 20 years, may generate some renewed interest in it. As background, this article reviews the history and development of the LMT. PMID- 16367437 TI - MAPS of the Harvard Yard. AB - The Make-A-Picture-Story (MAPS) Test was given to 14 undergraduates who were studied intensively at the Harvard Psychological Clinic by Henry Murray and his colleagues from 1959 to 1962. A case study of one of these subjects is presented to illustrate the use of the MAPS in drawing inferences about personality characteristics. PMID- 16367438 TI - Personality assessment and Native Americans. AB - Personality assessment services for Native Americans have been culturally inappropriate and historically underutilized as a consequence. A framework for personality assessment is presented including components of relevant cultural knowledge, assessment techniques, assessor characteristics, and relationship style. Emic and etic approaches are described as serving different assessment functions. Acculturation measures and awareness of acculturation effects on different instruments provide temporary palliatives. A biopsychosocial model for service-delivery with linkages to family, tribal, county, state, and federal resources would be desirable and potentially effective. Ameliorations in techniques and service-delivery cannot substitute for genuine professional commitment to recruitment and training of indigenous assessment service providers. PMID- 16367439 TI - The MCMI and DSM-III: a brief rejoinder to Million (1985). AB - Millon (1985) presented eight criticisms of the article by Widiger, Williams, Spitzer, and Frances (1985) on the MCMI as a measure of DSM-III. This article is a brief rejoinder. The major point we wish to make is that one should be cautious in interpreting the MCMI as a measure of DSM-III disorders because there has not yet been any research published on the relationship between the MCMI and DSM-III. We believe this position has not been refuted by Millon's critique. PMID- 16367440 TI - The Ascription of Responsibility Questionnaire: development and empirical extensions. AB - This article describes the development of the Ascription of Responsibility Questionnaire (ARQ). In Study 1, item analytic and factor analytic procedures were used, with a data set based on 251 subjects, to reduce an initial pool of 92 Likert Scale items to 41 items that were structured by four clear, independent ascription tendencies, or ways in which people ascribe responsibility. In Study 2, this factorial structure was confirmed on a sample of 403 subjects. From the responses in both studies, reliabilities of the four ascription of responsibility scales were assessed. On the basis of a pooled sample, norms were developed. Construct validity assessment was begun by correlating scores, in both studies, on the ascription of responsibility scales with a number of conceptually related standardized and semistandardized scales, as well as with a number of demographic and personological variables that might be related to attributional and attitudinal tendencies. These and further construct validation results obtained by comparative analytic methods are presented and discussed. PMID- 16367441 TI - Androgeny, flexibility, and individualism. AB - Questionnaire data from 195 college students were examined to clarify the relationships among masculinity, femininity, and androgyny and self-perceived flexibility and individualism. Analysis of variance (ANOVA) and hierarchical regression showed no evidence for unique positive "androgyny" effects, while higher masculinity was associated with flexibility and individualism, particularly among women. Contrary to expectations, "androgynous" men were more rigid and conforming, while the cross-typed were most flexible and independent. PMID- 16367442 TI - The circumplexity of Leary's Interpersonal Circle: a multidimensional scaling perspective. AB - To investigate the circulant structure of the 16 scales comprising the Interpersonal Checklist (ICL; Laforge & Suczek, 1955), intercorrelation matrices for mate and female undergraduate ICL scores were subjected to multidimentional scaling analysis. Results showed the following: (a) while the 16 ICL scales fall in a rough circular array, measurement gaps exist in the friendly-dominant and hostile-submissive quadrants; (b) sixteenths A-B-P and L-M-N were misarranged; (c) stress coefficients for a three-dimensional solution were in an acceptable range. Implications of these findings for future research and anchoring of the ICL to Kiesler's (1983) "1982 Interpersonal Circle" are discussed. PMID- 16367444 TI - Two objective measures of self-esteem. AB - Two scales were constructed to assess self-esteem, conceptualized as reflecting (a) feelings of competence and efficacy, and (b) perceived positive appraisal from significant others. To control for response bias a paired choice format was chosen for the items constructed. A buffer scale designed to measure social assertiveness was also included. Data were collected on three samples of high school boys. The item intercorrelations were subjected to principal component analyses followed by Varimax rotations. In each of the three analyses factors of Confidence, Popularity (Social Approval), and Social Assertiveness emerged. The revised self-esteem scales, each defined by 11 items, have been shown to have acceptable reliability and some concurrent validity based on correlations with the well-known Rosenberg Self-Esteem Scale. PMID- 16367445 TI - The Body Esteem Scale: a convergent and discriminant validity study. AB - This research was intended to further establish convergent and discriminant validity for a recent Body Esteem Scale (BES) which measures different dimensions of body satisfaction in young adults. One hundred and fifty-four male and 193 female undergraduates completed the BES, the Body Consciousness Questionnaire (BCQ), the Self-Esteem Scale, and several questions pertaining to exercise, food intake, and attractiveness. Good convergent and discriminant validity was demonstrated by the Male Upper Body Strength and Physical Condition subscales and by the female Weight Concern, Physical Condition, and Sexual Attractiveness subscales. PMID- 16367446 TI - Objectivism in information utilization: theory and measurement. AB - A self-report scale was constructed and validated that measures individual differences in objectivism--the tendency to base one's judgments and beliefs on empirical information and rational considerations. Validity data showed that, compared to people who score low on the Objectivism Scale, highly objective individuals enjoy thinking more, rely more on observable facts when making decisions, and place less emphasis on subjective and intuitive styles of decision making. Among graduate students in psychology, objectivism correlated positively with ratings of research-oriented careers, but negatively with ratings of mental health careers; also, highly objective students were more critical of nonobjective psychological assessment techniques and placed greater importance on research. Objectivism also predicted preferences for newspaper articles, college course selections, and the criteria respondents use when making decisions. PMID- 16367447 TI - Inconsistent responding to repeated MMPI items: is its major cause really carelessness? AB - Previous authors have recommended using the number of inconsistent responses given to repeated MMPI items as a measure of the carelessness with which an individual has responded to the test. Such a recommendation assumes that a respondent who reads the items carefully must necessarily give an identical response to both presentations of the same item. Contrary to this assumption of a single cause, three potential causes of inconsistency--maladjustment, carelessness, and indecision--were investigated among 202 normal undergraduates who responded to the whole MMPI twice. The results suggest that indecision may be a more important cause of inconsistency than carelessness, thereby calling into question the use of inconsistencies on repeated MMPI items as a "carelessness" scale. Implications regarding the processes involved in responding to personality items are discussed. PMID- 16367448 TI - Parental representations and orality. AB - One hundred ninety-three undergraduate male subjects were administered a Rorschach orality measure (Masling, Rabie, & Blondheim, 1967), and completed copies of Blatt, Wein, Chevron, and Quinlan's (1981) Parental Representations measure. A significant correlation between orality and quality of parental representations was found, with high oral subjects giving more negative descriptions of the mother than low orals. No relationship between orality and the conceptual level of parental descriptions was found. The implications of these findings are discussed in the context of psychodynamic formulations regarding the oral personality. PMID- 16367451 TI - Society for Personality Assessment, Inc. Bruno Klopfer Distinguished Contribution Award. Co-recipient, Stephen A. Appelbaum. PMID- 16367449 TI - The Separation-Individuation Test of Adolescence. AB - The procedures undergone to establish the validity of the Separation- Individuation Test of Adolescence (SITA) are described. The test consists of six scales designed to measure key dimensions of adolescent separation-individuation. Each scale was subjected to three stages of validation: theoretical-substantive, internal-structural, and external-criterion. The results of data with 305 adolescents are reported. Although particular modifications from the original form are planned, appreciative levels of validity were established for the SITA's various scales. PMID- 16367452 TI - Using early memories as a projective technique--the cognitive perceptual method. AB - This paper will present the cognitive-perceptual method of interpreting early memories together with the first published sequence interpretation of a set of early memories using this method. The latter is especially apt to present in this issue, considering Walter Ktopfer's stress on sequence interpretation of the Rorschach. Attention is also directed to the formation of a cross-disciplinary group that will investigate autobiographical memory in general and early memories in particular. PMID- 16367453 TI - A service-delivery paradigm for personality assessment. AB - This paper describes a human science approach to the delivery of the feedback component of assessment services. The feedback process is now central to a collaborative assessment that is integrated with intervention. Research has suggested that subject, situational, and assessor variables are relevant for feedback, but such research has been unsystematic. A service-delivery paradigm for feedback of assessment findings is described that includes four levels of increasing client participation: information, emotional contingencies, transfer of power, and functioning of an assessment system. Communication of assessment findings has been incomplete historically because motivation for feedback, resource accessibility for feedback utilization, and criteria for adequate feedback have been considered as unrelated ingredients. The components of Antonovsky's Sense of Coherence construct--meaningfulness, manageability and comprehensibility--provide one conceptualization for interaction of these ingredients. It is now feasible to do systematic research on assessment that includes feedback of assessment findings and the service-delivery process. PMID- 16367454 TI - Assessing offenders: dying for reprieve. AB - As is true for those who face death, the trauma of arrest and conviction for a crime and subsequent imprisonment takes the individual through a series of psychological stages, In the offender, these defenses are to some extent prescribed by the culture. The nature of and the person's adaptation to this process has not been previously recognized and argues for an assessment approach that emphasizes developmental rather than static formulations. Examination and treatment implications are discussed. PMID- 16367455 TI - The early development of the Freedman-Leary-Coffey Interpersonal System. PMID- 16367456 TI - Symposium: interpersonal circumplex models (1948-1983). PMID- 16367457 TI - Symposium: interpersonal circumplex models: 1948-1983 (commentary). PMID- 16367458 TI - Directions psychology takes. PMID- 16367459 TI - Humor and interpersonal attraction. AB - A humor test composed of cartoons, comic strips, and jokes was administered to 30 college couples (26 single, 4 married) who rated them for humor. Subjects also stated how much they loved and liked their partner, their probability of marrying the partner, and filled out Rubin's Liking and Love Scales. The hypotheses were that similarity of rating of the humorous stimuli would be associated with loving, liking, and predisposition to marry. Hypotheses were confirmed. PMID- 16367460 TI - The Sentence Completion Method--recent research. AB - The present article reviews recent work with the Sentence Completion Method (SCM) and examines the method's effectiveness as an assessment instrument. Studies in which the SCM was employed in testing hypotheses concerning personality and its development, as well as reports that deal with maladaptive functioning and psychopathology are covered. Also reviewed is some work that investigates the nature of the method itself. The overall conclusion is that the SCM is flexible, readily adaptable in various research and clinical settings, and to be recommended for continuing use and application. PMID- 16367461 TI - Personality assessment from the view of logical learning theory. AB - The gradual transition from clinical assessment to basic personality research in the author's professional development is reviewed, Thanks to the understanding and encouragement of Walter G. Klopfer the author found it possible to extend a line of teleological theorizing that has proven empirically viable for several years to date. This approach to human description is known as logical learning theory. The essentials of logical learning theory are reviewed and its application to personality assessment is highlighted. PMID- 16367462 TI - Psychological assessment and Miranda rights. AB - The Supreme Court has indicated that a criminal defendant must be advised of the right to remain silent and to be represented by an attorney, and a confession is ruled inadmissible if it is obtained without the knowledge of those rights. A psychologist may be placed in a position of being asked to advise the court as to the defendant's ability to comprehend and knowingly waive these rights. Three brief cases are presented in which meaningful comprehension was the issue, and a description is provided of the approach taken to responding to the question raised. PMID- 16367463 TI - Vivid persuasion in the courtroom. AB - When witnesses at a trial offer testimony that is vivid, it may be more persuasive than the same testimony offered in a pallid manner. In this paper we elucidate three categories of variables (inferential, attentional/memorial, and affective) that are likely to mediate the effects of the vividness of testimony on jury judgments. These variables are then used to explain discrepant findings among mock juror experiments investigating vividness effects. Finally, we discuss the implications of vividness effects for the legal system. PMID- 16367464 TI - What is the client's best interest? professional roles in the Criminal Justice System. PMID- 16367466 TI - Walter G. Klopfer, PhD. 1923-1985. PMID- 16367467 TI - Personality and self-actualizing profiles of assertive people. AB - This study tested the hypothesis that assertive and nonassertive people have different personality profiles and self-actualizing values. Results indicated that assertive and nonassertive groups differed significantly, in terms of their personality profiles based on the Personality Research Form, and their self actualization profiles based on the Personal Orientation Inventory, supporting the tested hypothesis. PMID- 16367470 TI - A brief inquiry into deviant answers to selected Grayson items of The Minnesota Multiphasic Personality Inventory. PMID- 16367471 TI - Evaluation of the BSRI masculine and feminine items using desirability and stereotype ratings. AB - This study empirically tested Bem's (1974) assumption that the BSRI Masculinity and Femininity scales measure sex-typed standards of desirable behavior for men and women in American society. The adequacy of items in the two scales was evaluated by Bem's (1974) criteria using two types of desirability and stereotype ratings. Results obtained in all the experimental conditions except one involving Bem's desirability instructions and Bem's rating scale did not support the tested assumption. Implications of these results for revising the BSRI scales were discussed. PMID- 16367472 TI - Pursuing and distancing: the construct and its measurement. AB - Although the concept of interpersonal pursuing and distancing has been introduced and used clinically, the lack of a reliable and valid measure has deterred its more formal investigation and relevance to personality theory. The Pursuing Distancing (P-D) Scale, a 92-item measure of the behavioral expression of these characteristics, was developed. Evidence for internal consistency as well as internal support of construct validity is presented and discussed. An 80-item revised scale is available from the authors and is currently being evaluated in a variety of external validation settings. PMID- 16367473 TI - The personality of chessplayers. PMID- 16367474 TI - Response to two presentations of the Sentence Completion Test of ego development. AB - This was a study of the differences among individuals on their performance on oral and written versions of the Washington University Sentence Completion Test of ego development. The study included 192 subjects from three age groups: College age, community residents, and retired people. No oral versus written differences were found in scores. Gender was the only factor to exhibit a consistently significant effect on Sentence Completion Test score differences. PMID- 16367475 TI - Orality and latency of volunteering to serve as experimental subjects: a replication. AB - Comparison of Rorschach orality scores for undergraduate subjects who signed up to participate in psychology experiments during the first (n = 217) and second (n = 87) halves of the semester revealed that early participators report a significantly (p = .03) greater number of oral Rorschach responses (X = 3.765) than late participators (X = 3.218). This result confirms an earlier serendipitous finding (Masling, O'Neill, & Jayne, 1981) that volunteering to participate in required experiments is not a chance event and can, in fact, be predicted using methods derived from psychoanalytic theory. Our finding supports the utility of psychoanalytic principles in predicting important aspects of nonlaboratory behavior. PMID- 16367476 TI - Construct validity of the Concern For Appropriateness scale. AB - Construct validity of the Concern For Appropriateness scale (CFA; Lennox & Wolfe, 1984), a measure of the tendency to adopt protective self-presentation styles, is examined using a multitrait-multimethod approach. Self-ratings on the CFA and ratings by suitemates constitute the two methods, and concern for appropriateness, interpersonal trust, and boredom susceptibility are assessed with each. Multitrait-multimethod analysis shows that the CFA scale exhibits convergent validity against peer ratings and discriminant validity against boredom susceptibility, the only other variable demonstrating convergence. Results indicate that the CFA exhibits construct validity and might therefore prove useful as an operationalization for self-presentation tendencies. PMID- 16367477 TI - Discriminant measures for desperate love. AB - Viewing desperate love as marked by urgency, neediness, dependence, a great need for reciprocation, indealization, and affective extremes, personality traits leading to desperate love were examined. A sample of 251 undergraduates, in desperate love, nondesperate love, and random groups, completed several questionnaires examining characteristic qualities of self and important others and attitudes toward love (Desperate Love Scale, Marlowe-Crowne Social Desirability Scale, Knox-Sporakowski Attitudes Toward Love Scale, Characteristic Qualities Questionnaire). Findings support both the primary hypothesis that people can be differentiated in their experience of desperate love based upon significantly different responses when describing characteristic qualities of self and important others and the secondary hypothesis that those tending toward desperate love have a more romantic attitude toward love. The study can be taken as an indication that latent qualities, as well as manifest behavior, differentiate someone who experiences desperate love. PMID- 16367479 TI - The reliability of the Thematic Apperception Test. AB - Controversy over the TAT's reliability may stem largely from the mis-application of traditional psychometric measures, which are inappropriate to this test. The TAT is implicitly based on a multiple regression model, for which coefficient alpha is not appropriate. Also, test-retest correlations may be adversely affected by the standard instructions to write a "creative" story. In a test retest study, 47 high school students retook the TAT after a year with instructions designed to break the implicit set to produce a new and different story from that previously written. The test-retest correlations were r = .48 (need for affiliation) and .56 (need for intimacy), or approximately the same as those for, e.g., the MMPI, 16PF, and CPI, It was demonstrated that this high stability over time was not due to subjects' recalling and repeating previous responses. Finally, it was shown that alpha considerably underestimated the test retest reliability, contrary to assumptions of classical psychometrics. PMID- 16367480 TI - MMPI Scales for measuring Eysenck's personality factors. AB - Four new MMPI Scales were constructed to identify Eysenck's personality factors of psychoticism, extraversion, and neuroticism as well as a lie score. The scales showed good construct validities against the Eysenck Personality Questionnaire and the Friedman Overlap Scales developed from items in the MMPI. The study also supported the construct validity of the Friedman overlap scales. PMID- 16367481 TI - A modification in scoring the ABC Scale. AB - The scoring procedures for the Articulation of the Body-Concept (ABC) Scale to use with three-, four- and five-year-old children were modified to determine their degree of field-dependence-independence. A total of 300 children, who obtained a score of one or better on the ABC Scale, were administered the Preschool Embedded Figures Test (PEFT) and the ABC Scale. The ABC Scale was scored using its scoring procedures and the scoring procedures for the Goodenough Harris Drawing Test (GHDT). Scores on the PEFT, ABC Scale and GHDT were correlated with each other. Correlations among all cognitive measures ranged from .88 to .99, which were high and statistically significant (p < .0001). Therefore, the scoring procedures for the GHDT can be substituted in scoring the ABC Scale in assessing young children's field-dependence-independence. PMID- 16367482 TI - Differential correlates of irrational belief. AB - One hundred and ten students completed a battery which included Jones' Irrational Belief Test (IBT), and measures of subjective well-being, affect, alienation, personality, goal characteristics, and goat attainment. Correlations and canonical composites showed a general relationship between irrational beliefs and emotional difficulty, but the specific pattern differed for subgroups of IBT scales. Correlations supported the construct validity of IBT scales, but suggested they also reflect motives, conflicts, affects and personality characteristics. Interdependence of these processes was hypothesized and use of subscore composites rather than total IBT scores was suggested. PMID- 16367483 TI - Age, gender, and interpersonal behavior development using the FIRO-BC. AB - It has been argued that many human behaviors follow predictable developmental patterns, or stages. The FIRO-BC was given to 9- through 13-year-old children (n = 282) to test for a stage-like progression in interpersonal behavior development. The data presented here failed to evidence an age-related progression. However, gender differences did appear in 11-year-olds and increased dramatically by age 13. Geographical differences were found in that the present data differed significantly from the normative data presented in the FIRO manual. The data also support the notion that, at least within the southern population tested. interpersonal behavior may be more related to puberty than has been previously supposed. PMID- 16367484 TI - A short scale to measure self-righteousness. AB - Two studies were conducted which created and tested a scale to measure self righteousness. Self-righteousness was defined as the conviction that one's behaviors or beliefs are correct, especially in contrast to alternate behaviors or beliefs. In the first study, a 4-item scale to measure general self righteousness was derived which demonstrated adequate levels of internal consistency and was related to dogmatism and ambiguity intolerance. In the second study, the items were worded to be specific to running a race. This study succeeded in demonstrating that the scale possessed predictive validity. This study also demonstrated the usefulness of rewording the general items to make them specific about the content of self-righteousness. PMID- 16367485 TI - Efficiency of local versus standard MMPI norms: a comment. AB - This note shows that evidence considered by Wooten (1984) to support claims of diagnostic superiority of standard over local MMPI norms, may mainly reflect the fact that a large proportion of the group tested by Wooten's consisted of persons with emotional, behavioral problems. Reevaluation of Wooten's data suggests a range of conditions under which Wooten's diagnostic criterion used with local norms is superior to the same criterion used with standard norms, and another range of conditions under which this criterion used with either local or standard norms is inferior to base rates. PMID- 16367486 TI - Measuring activism regarding the nuclear arms race. AB - A 14-item questionnaire measure of activism regarding the nuclear arms race is described. Scale development analyses were conducted on 227 subjects, and internal consistency reliabilities ranged from .56 to .91 for scales assessing four aspects of activism on this issue. High correlations were obtained between these brief scales and corresponding measures derived from a 50-item long form. Responses of five subject groups were also compared: workers in defense industries, participants in a Republican party meeting, graduate students in psychology, religious teachers, and peace activists. Applications of this measure for study of the behavioral component of responses to the nuclear arms race are discussed. PMID- 16367490 TI - Rorschach content interpretation in post-traumatic stress disorders: a reply to Kowitt. PMID- 16367491 TI - Interpersonal affect in Thematic Apperception Test responses: a scoring system. AB - Affect toward "parental," "spousal," and "nonspousal" figures in TAT responses was evaluated. Positive and negative scores ranging from +2 to -2 were assigned to each story. Norms were based on frequent responses stimulus-pull and quality Of responses Test-retest rendered .88 similarity in affect and 86% in orientational others categories. Interscorer's agreements on affective scores ranged from .93 to .97 and interpersonal categories ranged from 96% to 98%. Higher positive "spousal" than "parental" and "nonspousal" affect was hypothesized to associate with marital adjustment. Thirty married Caucasian middle-class couples were individually interviewed. Cards 1, 2, 4, 6 GF, 10 13 FM and 16 as well as the Dyadic. Adjustment Scale were administered. Results, significant at .001 level, were discussed in terms of development in interpersonal choices. PMID- 16367492 TI - A literature review of subtle-obvious items on the MMPI. AB - This article chronologically reviews the literature relating to subtle and obvious items of the Minnesota Multiphasic Personality Inventory. Wiener's original development of subtle and obvious subscales, Seeman's support for the concept of subtlety, and Duffs arguments concerning empirical versus face validity are discussed. Later research exploring demand characteristics are then reviewed, such as the paradoxical relationships under differing instruction sets and population issues. The work from Auburn University researchers on the development of a 5-point subtlety scale, the interaction with personality factors, and individual scale research is critiqued. PMID- 16367493 TI - The Satisfaction With Life Scale. AB - This article reports the development and validation of a scale to measure global life satisfaction, the Satisfaction With Life Scale (SWLS). Among the various components of subjective well-being, the SWLS is narrowly focused to assess global life satisfaction and does not tap related constructs such as positive affect or loneliness. The SWLS is shown to have favorable psychometric properties, including high internal consistency and high temporal reliability. Scores on the SWLS correlate moderately to highly with other measures of subjective well-being, and correlate predictably with specific personality characteristics. It is noted that the SWLS is Suited for use with different age groups, and other potential uses of the scale are discussed. PMID- 16367494 TI - Assessing mood with affect-sensitive tasks. AB - It is hypothesized that when an individual experiences a change in mood there are concomitant changes in performance at cognitive and psychomotor tasks. The present study attempts to identify tasks which are mood-sensitive. The nature of affect-sensitive tasks will yield information about how mood influences behavior. Further, to the extent that relationships between mood and affect-sensitive tasks are found, clients may be motivated to change their mood by a knowledge of expectable improvements in performance. Ten tasks and a mood-adjective checklist were administered to 106 subjects. Four of the tasks showed statistically significant relationships with self-reported mood. Results were promising for further investigations of affect-sensitive tasks. PMID- 16367495 TI - Human versus computer interviewing. AB - Responses to counterbalanced human and computer interviews were examined for Gordon's Survey of Interpersonal Values to determine whether such responses would differ. Of an initial 72 male and female subjects, 52 completed both interviews scheduled one week apart. A 3-way MANOVA revealed significant response differences between interviewer type. However, neither significant sex or interviewer order differences, nor significant interactions were found. The results suggest that the interpretive validity of responses obtained in a computer interview may be threatened. The authors additionally suggest that the observed differences may be due to an interaction between interviewer type and test anxiety level. PMID- 16367499 TI - Megatrends in personality assessment: toward a human science professional psychology. AB - Societal megatrends have assessment counterparts--superempiricism, biopsychosocial perspective, person-centering, population-specificity, self assessment--which are used to examine the contemporary status of professional psychology. A human science approach to research, training, and practice can exemplify a profession that acknowledges these megatrends. The nurture of competent professional psychologists requires a reinvestment in Boulder Model ideology, a human science that is shared with consumers on the basis of values which communicate caring and understanding. PMID- 16367500 TI - The architect as person: a Rorschach image. AB - Analysis of the Rorschachs of 40 eminent male architects collected in 1959 revealed a highly consistent personality profile. The picture that emerges is of a person who appears to be ambitious, highly sensitive, intellectually and emotionally energetic, and reasonably well adjusted socially. His thinking is well oriented in reality. He prefers large scale conceptualizing to mundane 'concerns. There is evidence of conflict, tension, and a concern with emotional control. Feminine and exhibitionistic interests are exceptionally high and sources of inspiration appear to reside in unresolved sexual conflicts. The attitude toward the world is positive. The eminent male American architect at the turn of the Sixties believed in his own ability to create a truly superior environment. His conscious concerns were with creation rather than with self confrontation. PMID- 16367501 TI - Personality and "success" among a selected group of lawyers. AB - As part of a larger study, eleven subjects--male, Caucasian, native-born, early 70s, lawyers, life-long (since 1922) participants in the Terman Study of the Gifted--were interviewed. From the edited tapes, seven qualified raters evaluated Various measures of life success and completed a 100-item Q-sort for all subjects. In general, the very highest successes were related to contentment, self-confidence, openness and spontaneity, a wide range of cultural interests, and relative freedom from pervasive feelings of hostility, irritability and dissatisfaction. The general results are corroborated by Terman Study data available for these same subjects for years 1922, 1927, and 1940. Some implications of these findings for child-rearing, marriage and the possible governance of one's life are explored. PMID- 16367502 TI - Individual differences in hedonic capacity. AB - Hedonic capacity--an individuals ability to experience pleasurable affect--is important, not Only in accounting for the development of psychopathology but in understanding normal personality as well. In this article, the development and validation of three scales of hedonic capacity are described. One scale consists of MMPI items, a second consists of CPI items, and the third combines items from both inventories. These scales demonstrated acceptable internal consistency and test-retest reliability. Initial support for the construct validity of the scales was also obtained. Because they have been constructed from the MMPI and CPI, the scales described in this article can be used to address important questions in research on schizophrenia, depression, and normal personality with a wide variety of data that has already been collected. PMID- 16367503 TI - Anality and Type A coronary-prone behavior pattern. AB - The extent of overlap between the descriptively similar Type A coronary-prone behavior pattern and the psychoanalytic anal character was investigated empirically. Pearson product-moment correlations were computed on the scores of three samples of male introductory psychology, students on the Kline Ai3 anality scale and the Jenkins Activity Survey. As predicted three positive correlations resulted, two significant and the third a trend with an overall combined significant probability for the three samples (p < .005). The implications of these findings for the Type A literature are discussed. PMID- 16367504 TI - A modification of the Fenigstein, Scheier, and Buss Self-Consciousness Scales. AB - Confirmatory factor analysis was employed to examine the structure of the Fenigstein, Scheier, and Buss (1975) Self-consciousness Scales. Several weaknesses were identified and modifications were made to improve the representation of the underlying structure of the data. Five items were found to be relatively unreliable and were dropped. Two separate but correlated private self-consciousness dimensions, self-reflectiveness and internal state awareness, were identified. The resulting four-factor self-consciousness model fit the data more adequately than the three-factor model of Fenigstein et al. PMID- 16367505 TI - Validation of an automated 16PF administration procedure. AB - Examined the effects of a computer-based administration procedure on reliability and validity of a personality questionnaire. Utilizing a counterbalanced repeated measures design, Form A of the 16PF was administered to 80 undergraduates who had been randomly assigned to either computer-computer, computer-booklet, booklet computer, or booklet-booklet administration conditions. Subjects also completed a self-report anxiety measure before and after each administration of the 16PF. Multivariate analyses revealed no significant differences in score reliability, validity, or self-reported anxiety among the four administration conditions. Small yet significant differences were noted with respect to ratings of the assessment experience. Subjects in the two groups which were exposed to both administration formats rated the computer procedure as more positive in comparison to the booklet format. Due to the student sample, replication of these findings with treatment-seeking clients is needed. PMID- 16367508 TI - Rorschach object relations and fantasy themes incorrectly scored as determinants. AB - Determinants are perceptual features of inkblots, but movement, reflections, pairs, and subdivisions of shading, scored as determinants, actually represent associative elaborations. Such misclassification of determinants obfuscates the response process and makes it difficult to evaluate the validity of inferences. To the extent that these scores are useful, they can be improved upon by fully developed content analysis. Movement, reflections, and pairs can best be understood as manifestations of object relations content themes, as scored in systems developed by Blatt, Krohn & Mayman, and Urist. Determinant scores for reflections and pairs should be eliminated. Shading should not be subdivided into texture, vista, and diffuse shading, since these are content themes without clear psychological significance or clear empirical support. PMID- 16367509 TI - Personality correlates of the Matching Familiar Figures Test in adults. AB - The Matching Familiar Figures Test (MFFT) measures a dimension of cognitive style known as reflection-impulsivity. Interpretations of the MFFT sometimes supplement its cognitive implications with one of two personality models, impulsivity and/or ego-resilience. Both models have received some research support from studies that used preadolescent subjects. The applicability of these two personality models to adults was evaluated in this research. In three studies on college students, one personality characteristic within the ego-resilience position, independent oriented achievement, was found to differentiate between impulsive and reflective scorers on the MFFT. No support was found for the impulsivity position. Implications were drawn for the influence of personality on the MFFT. PMID- 16367510 TI - An MMPI-based behavior descriptor/personality trait list. AB - Descriptive terms associated with elevated MMPI scales were abstracted from 10 MMPI reference works. Those corresponding to the 300-word Gough Adjective Check List were retained to construct a new behavior descriptor/personality trait list. PMID- 16367511 TI - A revised measure of approval motivation and its relationship to social desirability. AB - Existing measures of approval motivation attempt to assess this dimension of personality indirectly through evaluation of socially desirable response tendencies. The Martin-Larsen Approval Motivation (MLAM) Scale takes a more direct approach by focusing on individual differences in need for social approval. Since the original version of the MLAM scale is subject to acquiescence response bias, fully balanced and partially balanced scales were created to minimize this response style. Results indicated that the revised measures arc equivalent to the original version and that balancing affects the magnitude but not the pattern of correlates with other scales. Comparisons with the Marlowe Crowne Social Desirability (MCSD) Scale revealed a divergent pattern of correlates with several measures of personality suggesting a basic difference between the MLAM and MCSD in their conceptualization of approval motivation. PMID- 16367512 TI - Sex differences in affective responses as evidenced on the Bene-Anthony Test. AB - This study investigates changes in a child's perception of the relationship with male and female parents during the course of the critical ages of 5, 6, 7, and 8. Correspondingly, since the paper concerns itself with nonpathological normal children, it also provides normative data on the Bene-Anthony Family Relations Test. Findings indicate variable gender and age differences in the feelings experienced by children aged 5, 6, 7, and 8 towards male and female parents. There are differences between male and female children in their attitudes toward mother and father, and there are significant changes within the age ranks. Sweeping norms which do not take age and gender into account are inappropriate on the basis of our findings. PMID- 16367513 TI - A multidimensional loneliness measure: the Loneliness Rating Scale (LRS). AB - Interest in the topic of loneliness has resulted in the development of a number of global, unidimensional instruments used for measuring current levels of loneliness. A new, multidimensional scale, the Loneliness Rating Scale (LRS), is introduced. This 40-item scale assesses the frequency and intensity of particular affects reported by lonely individuals. Results of a factor analytic procedure suggest that the LRS contains four affective dimensions: Depletion, Isolation, Agitation, and Dejection. The theoretical implications of these results are presented, along with information concerning the usage and scoring of the LRS. PMID- 16367514 TI - The measurement of generalized self-efficacy: a study of construct validity. PMID- 16367515 TI - The discrepancy between private and public selves: public self-consciousness and its correlates. PMID- 16367517 TI - Some test correlates of high level creativity in architects. AB - The Rorschachs of 40 eminent architects, intensively studied at the Institute of Personality Assessment and Research (California) in 1959, were analyzed in terms of styles of responding (cognitive-affective controls). Level of creative talent (assessed by a competent jury of peers) was not related to styles of responding but the Rorschach data give us reason to believe that subjects with high degrees of repressive control may be more limited in creative expression. Qualitative analysis of Rorschach content suggests drive level is related to creative excellence. PMID- 16367518 TI - Exploring fantasies of TAT reliability. AB - It has been argued that low TAT stability estimates are an artifact of retest instructions to be original and creative. This view was supported in a recent study by Winter and Stewart (1977) who found that test-retest reliability was higher when subjects were told on the second occasion to write the same stories than when they were told to write different stories. This study attempted to replicate Winter and Stewart's results and also to determine whether the similarity of stories did vary as hypothesized. The replication was only partially successful. More importantly, it was concluded that while instructions to write similar stories did improve reliability, the increase reflected only subjects' memory capabilities and not greater stability in motive expression. While true TAT stability may still be commonly underestimated, altering retest instructions to improve reliability does not appear to be the solution. PMID- 16367519 TI - Warrior, general, and president: Dwight David Eisenhower and his earliest recollections. AB - The earliest recollections of Dwight David Eisenhower, supreme military commander of allied forces during World War II and 34th president of the United States, are analyzed to provide a deeper understanding of the man and his motives. The analysis emphasizes a cognitive/perceptual method of early memory interpretation presented by Bruhn and Last (1982) in a theoretical paper. PMID- 16367520 TI - The measurement of role taking in young adults. AB - Sociometry is presented as an alternative way of measuring role taking, specifically, an alternative to an often used projective measure, the Role-Taking Task (RTT). Study 1 assesses role taking in 161 college students. In this study, a sociometric measure, the Peer Role-Taking Questionnaire (PRTQ) was shown to correlate with class year, group status, social choice, and friendship. In contrast, the RTT correlated weakly with grade point average and some personality test measures. In Study 2, the PRTQ, and not the RIT, was associated with interaction skills in a sample of 39 college nursing students. The PRTQ and RTT did not correlate with each other in either study. The validity and reliability of these two role-taking measures are discussed. It is suggested that researchers need to examine the adequacy of the role-taking measures they use. PMID- 16367521 TI - Barnum meets the computer: a critical test. AB - The effects of type of assessment device (computer vs. human) on subjects' acceptance of a favorable or unfavorable personality feedback were investigated. Forty-four French-speaking subjects were administered a fictitious personality questionnaire and were asked to evaluate how appropriate the personality feedback was as a description of themselves and people in general. Contrary to predictions, there was no interaction between the type of assessment device and the favorability factors. Two previous findings were nevertheless replicated: (a) in all conditions the favorable description was better accepted than the unfavorable one and, (b) subjects rated the favorable description as being more appropriate for themselves than for people in general. PMID- 16367522 TI - Application of the consensus Rorschach to couples. AB - The consensus method of administering the Rorschach is described and the literature reviewed. The consensus method is viewed as an adaptation of the Rorschach as a form of behavioral assessment, especially as regards couple interaction. Materials on two couples are presented as illustrations. PMID- 16367525 TI - Item validity of the Myers-Briggs Type Indicator. AB - The present study presents a brief summary of four extensive psychometric analyses of the Myers-Briggs Type Indicator (MBTI) items. Positive empirical evidence supports the MBTI item validity. However, several measurement issues on item construction were raised to caution the future users. PMID- 16367526 TI - Inattention to history yields faulty conceptions of Nazi leaders. AB - Harrower (1976) as well as Miale and Seizer (1975) partly base their analyses of the Nazi leadership on the contention that Hjalmar Schacht, Minister of Economics for the Third Reich, was a typical Nazi. Those analyses are called into question by the contention that Schacht was not a Nazi at all. Further, it is argued that the failure to understand Schacht is not the only misjudgment made by the two sets of authors. The same literature that suggests Schacht was not a Nazi indicates that the theses of Harrower and Miale-Selzer are too simple. PMID- 16367527 TI - MMPI Scale Five: its meaning, or lack thereof. AB - As a widely used test of personality and psychopathology, the MMPI play an important part in making decisions that affect people's lives. The addition of "cookbooks" and, more recently, computer-generated output based on MMPI scale scores provide even the most unsophisticated user with what appear to be credible and valid interpretations of scale and profile variations. A comprehensive review of the literature on MMPI Scale Five (MJ) calls into question generalizations arising from scores on the scale. Empirical research provides no consistent meaning for any score level on the scale. Attempts to map the structure of the scale result in numerous factors and account for, at best, 30% of the variance. Discussion focuses on the shortcomings of the scale as well as the assumptions on which it was developed. Conclusions suggest that the concept of sex difference itself is inappropriately defined and establishes artificial boundaries. PMID- 16367528 TI - Factor analysis and construct validity of the Narcissistic Personality Inventory. AB - Three studies are reported which provide evidence for the validity of the Narcissistic Personality Inventory (NPI). Factor analysis of the NPI in Study 1 revealed four factors which were labelled: Exploitativeness/Entitlement, Leadership/Authority, Superiority/Arrogance, and Self-absorption/Self-admiration. In Study 2, scores on the NPI were correlated with basic dimensions of personality, and with relevant self-variables. Narcissism scores were positively related to dominance, exhibitionism, extraversion, self-esteem, and self monitoring, among others; and negatively related to abasement, deference, and social anxiety, among others. Correlations between the NPI factors and personality variables are also examined. In Study 3, peer ratings of narcissism were obtained and it was found that these were strongly related to scores on the NPI. Taken together, the three studies provide considerable evidence for the construct validity of the NPI, and avenues for future research are suggested. PMID- 16367529 TI - Narcissism and empathy: validity evidence for the Narcissistic Personality Inventory. AB - Three empathy questionnaires were correlated with scores on the Narcissistic Personality Inventory (NPI), and the predicted negative relationship was observed in two of the three cases. In addition, the Exploitiveness/Entitlement (E/E) subscale of the NPI was inversely related to all three empathy scales and directly related to another hypothesized measure of narcissism. Unexpected negative correlations between the E/E subscale and social desirability scales were interpreted as evidence of E/E subscale sensitivity to the manipulative dimension of narcissistic relationship styles. These data further support the construct validity of the NPI in general and of the Exploitiveness/Entitlement subscale in particular. PMID- 16367530 TI - The efficient assessment of need for cognition. AB - A short form for assessing individual differences in need for cognition is described. PMID- 16367531 TI - Factor analyses of two 'attitude toward gender role' questionnaire. AB - Two questionnaires which purport to assess attitudes toward gender role, the Maferr Inventory of Feminine Values and the Sex Stereotype Questionnaire, were submitted to a factor analysis. While each analysis of the five response set items yielded interpretable factors with unequivocal item scores, the obtained factors are not consistent with the a priori assignment of items to "groupings" suggested by the manuals for these instruments. The implications of using questionnaires with questionable psychometric properties is discussed. PMID- 16367532 TI - The development of an Assertiveness Self-Report Inventory. AB - The development of a self-report measure of assertiveness is described. The Assertiveness Self-Report Inventory (ASRI) was devised with concern for endorsement frequency, internal consistency, and independence from social desirability response bias. The scale's reliability and convergent and discriminant validity are adequate. Further tests of validity support the use of the scale as a measure that taps behavior related to the construct of assertiveness. PMID- 16367537 TI - A comparison of English and Spanish (Nunez) translation of the MMPI. AB - As more Hispanics seek help in mental health centers, the need arises for a reliable Spanish translation of the MMPI. This present research is a preliminary attempt to study the validity of the Nunez translation of the MMPI, the translation most widely used in Latin America. This translation and the English MMPI were administered to a volunteer sample of 18 teenaged bilingual Hispanic women. A split-plot factorial ANOVA with one between-subjects factor(form of test) was used to analyze the K-corrected T scorns from each scale separately. On scales F, K, Hs, Pa,.and Sc, the Spanish means were significantly (p < .05) higher than the English means. Order of administration had no significant effect, and there was no significant interaction effect. These findings demonstrate that the Nunez translation cannot be used interchangeably with the English form of the MMPI. PMID- 16367538 TI - Effects of instructions and rating scales on item selection for the BSRI scales. AB - Walkup and Abbott (1978) stated that Edwards and Ashworth's (1977) failure to replicate Bem's (1974) selection of items for the Masculinity and Femininity Scales of the Bern Sex Role Inventory (BSRI) may be attributed to differences in the instructions and anchored rating scales used in the two studies. The present study tested the hypothesis that presence of various interaction effects involving instructions and rating scales would influence the acceptability of items for the BSRI Masculinity and Femininity Scales. Results based on the evaluation of individual items by Bem's item selection criteria in each of the four experimental conditions obtained by systematically manipulating two instructions (Bem's and Edwards' instructions) and two rating scales (Bem's and Edwards' rating scales) and also those based on the analysis of variance of item mean desirability ratings from the four experimental conditions supported the hypothesis. PMID- 16367539 TI - Creativity profile of university and conservatory dance students. AB - What are dance students' creative potential, Special personality characteristics and preferences for high energy activities? Using standardized and original paper and pencil instruments, comparisons were made among dance students in university and conservatory settings, college students and art students. The results demonstrated that dance students are creative, show strong preference for dynamic figures, and have distinct personality characteristics. The university dance students differed from the conservatory students in several important ways. PMID- 16367540 TI - Students' perception of actual, trivial, and inaccurate personality feedback. AB - Examined the ability of college students at two levels of defensiveness to discriminate among three types of personality feedback: actual personality feedback based on the student's test scores, trivial feedback which is generally true of all students, and inaccurate feedback which was the exact opposite of the student's actual test scores. The students could reliably discriminate the three types of feedback on the dimensions of uniqueness, usefulness, accuracy, and as a source of new information about themselves. The results are discussed in the context of the Barnum literature to the effect that students can offer valid perceptions of personality descriptions of themselves. PMID- 16367547 TI - The Family Responsibility Index: a behavioral measure of marital work allocation. AB - An instrument was developed to measure behaviorally the division of family-role responsibilities between spouses. Currently available marital role inventories, attitudinal scales and sex role surveys represent inadequate or inappropriate methods for measuring actual performance. The Family Responsibility Index (FRI) was designed to assess a spouse's degree of behavioral participation in 54 specific tasks divided among 10 areas of typical family responsibilities. A balance of traditionally masculine and traditionally feminine tasks was approximated. Sixty-one dual career family couples completed the instrument in two separate administrations. Results showed moderately high reliability of report between spouses and acceptable concurrent validity. PMID- 16367548 TI - A metric for integrating theoretically related but statistically uncorrelated constructs. AB - When considering psychological constructs in a pairwise fashion, a 2 x 2 category system, based on median splits, is often adopted for analytic convenience. If theoretical focus is on an individual's integration of the two constructs, then categorical procedures are subject to information loss and unreliability in high density regions of the data. A continuous metric is proposed which ameliorates these problems and is suitable for correlational analyses. The construct of psychological androgyny is used for illustrative purposes. PMID- 16367549 TI - The relationship between two measures of self-actualization. AB - The Personal Orientation Dimensions have been introduced as a refinement and extension of concepts of self-actualizing first measured by the Personal Orientation Inventory. The two inventories are theoretically and empirically compared and contrasted. Overall, there is little to favor the POD. It adds little, if anything, to what the POI provides. The recommendation is that users keep using the POI and not use the POD. PMID- 16367550 TI - Comment on 'Measuring Machiavellianism with Mach V: a psychometric investigation'. AB - A recent article (Shea & Beatty, 1983) attacks the reliability, triadic-choice format, and validity of the Mach V. The issues of meaningful versus statistical criteria, differing perspectives of bias, and reliability-validity independence are raised in challenge. Although the Mach V may have some construct validity problems, Shea and Beatty's call for a new measure of Machiavellionism is not supported by their article. PMID- 16367551 TI - A multidimensional assessment of the validity of the Personal Orientation Inventory. AB - The present study investigated the validity of a global measure of self actualization derived from the POI. The POI Inner-directed scale was related to numerous objective indices of self-actualizing characteristics (e.g., self esteem, rigidity, empathy). Bivariate and multivariate correlational techniques presented a consistent picture of the self-actualizing dimensions tapped by the POI Id scale (e.g., self-esteem) as well as the self-actualizing characteristics unrelated to Id scale scores (e.g., machiavellianism). The results help to clarify the specific nature of the attributes measured by the Id scale, partially supporting its construct validity as an omnibus measure of actualizing tendencies. Finally, a new, face-valid measure of self-actualization is introduced which proved as valid as the POI as an index of self-actualization, with a considerable gain in parsimony. PMID- 16367552 TI - Personality assessment: practice and teaching for the next decade. AB - This paper describes some loci for personality assessment practices in the next decade. There are new domains for description of Personality Disorders, stress and coping skills, and individual modernity and acculturation. An emphasis on psychological health assessment is consistent with primary and secondary intervention. A focus on competence and awareness of environmental demands leads to a rationale for assessment components of a psychological health evaluation. Education for the general public and training for professional assessors are relevant teaching goals. Competence, fairness, and an adequate repertoire of tools are requisite for professionals. PMID- 16367553 TI - Rorschach reliability: toward a training model for interscorer agreement. AB - Little or no research has been done with Rorschach Scoring Systems to standardize instructional materials and methods which would yield high interscorer agreement by the conclusion of a time-limited course of study in a classroom setting. Clinical applications and research with the Rorschach would benefit from training methods which yield high interscorer reliability. Training materials based on the Perceptanalytic system were standardized on 250 graduate level psychology students over an 11-year span of time, using one class per year. The final training materials were replicated using three classes (n = 58) to test the stability of the system. Data analysis on interscorer agreement involved 20,042 comparisons of pairs of scores and summaries at two levels of difficulty: Intermediate and Advanced. Overall interscorer agreement was consistently high for all replications. PMID- 16367554 TI - Relationships between Rorschach variables as relevant to the interpretation of structural data. AB - The protocols of 100 adult nonpatients who had been administered the Rorschach twice in a long term temporal consistency study were used in this study. Separate intercorrelational matrices were calculated for each testing with R partialed. Means of the pairs of partial correlations were calculated, the square root of which is considered the best available estimate of the correlation between each pair of variables. Fifteen of the 120 partial correlations are considered to be significant. The findings are discussed in terms of their potential relevance for configurational interpretation of structural data. PMID- 16367555 TI - A method for coping with overall productivity when making between-group category comparisons. AB - A straightforward method for dealing with the effect of response productivity when comparing individual scoring categories between groups was presented along with an example. It was contended that percentage comparisons based on group data can circumvent the reliability problems associated with percentage scores derived from single protocols. Other problems connected with ratio and percentage scores were discussed. PMID- 16367556 TI - Studies of Jungian typology: III. Personality and marriage. AB - Jung's theory of psychological types provided the conceptual framework for a study of personality characteristics implicated in marital relationships. Partners in 20 couples completed the Myers-Briggs Type Indicator and a series of open-ended questions describing their marital relationships. As predicted, (a) type similarity was more characteristic than type dissimilarity, particularly on sensation-intuition and thinking-feeling preferences, (b) sensation-intuition preferences were reflected in perceptions of qualities valued in self and spouse, (c) thinking-feeling preferences were significantly related to reported satisfaction in marriage, and (d) intuitives were more open to changes in the relationship than were sensing types. Marital problems were highly diverse, and were not related in any clear way to type patterns in individuals or dyads. PMID- 16367558 TI - Measurement of intimacy attitudes: the Intimacy Attitude Scale--revisited. AB - The study evaluated the reliability and validity of a revised version of the Intimacy Attitude Scale originally developed by Amidon. The Cronbach alpha measure of internal consistency showed alpha values between .78 and .86 in five different groups. The test-retest correlation was .57 which improved to .84 after exclusion of three extreme scores. Validity was examined by correlating with several variables. It was concluded that the revised Intimacy Attitude Scale is a reliable instrument with Some degree of Validity for use in research, diagnosis, and training. PMID- 16367559 TI - Typical vs. maximal measures of personological variables. AB - Recent research has focused on various ways in which self-report personality measures might be improved. The present investigation continues this line of research by studying the effects of typical-maximal ratings, trait-consistency, familiarity, and interpersonal liking on self-reports of four personological variables behavior aggression, trait-aggression, trait-dominance, and trait friendliness. The results revealed that maximal ratings were clearly superior to typical ratings. Furthermore, maximal ratings attenuated the effects of the moderator variable of trait-consistency. Subject-peer familiarity and interpersonal liking also demonstrated significant effects. Implications for personality assessment are discussed. PMID- 16367565 TI - The reliability and concurrent validity of alternative methods for assessing ego development. AB - This study demonstrates that an unstructured interview (INT) and the Thematic Apperception Test (TAT) are suitable alternatives to Loevinger's Sentence Completion Test of Ego Development. Seventy subjects were solicited from six groups varying widely with respect to age and education level. Each subject was asked to complete the SCI, an INT and the TAT. Two raters using Loevinger and Wessler's self-training exercises and Loevinger, Wessler, and Redmore's scoring manual rated subjects' responses to each instrument. Reliability was found between raters and concurrent validity between instruments. Subjects scoring high on the SCT were found to score higher on the INT and TAT. PMID- 16367566 TI - Peer ratings of males and females possessing different sex-role identities. AB - Two investigations were conducted to explore peer ratings of males and females exhibiting different sex roles. In the first study, 160 males and females representing four sex-role groups were rated by close, same sex friends on Gough's Adjective Check List. The results indicated that for both males and females, the four sex-role groups were perceived differently by their friends. I n the second investigation, peer rated adjectives which differentiated between the sex-role groups in Study I were evaluated on a positive/negative dimension; self-rated adjectives differentiating between the four sex-role groups in an earlier study (Baucom, 1980) were evaluated similarly. The findings showed that the peer-rated and self-rated adjectives which differentiated androgynous males and females from the other sex-role groups were viewed more positively than adjectives describing any other group: peer-rated and self-rated adjectives describing undifferentiated persons were consistently viewed negatively. Significant differences between the evaluations of the peer-rated and self-rated adjectives suggest that conclusions about the effects of sex roles are likely to vary, depending upon whether self-report inventories or peer ratings are investigated. Cautions are presented regarding generalizations from only one method of measurement. PMID- 16367567 TI - Factor analysis of the interpersonal trust scale with a noncollege population. AB - This study tested the invariance of the factorial structure of Rotter's Interpersonal Trust Scale (ITS) in a noncollege population. Exploratory factor analysis of the responses of 214 volunteers yielded three factors interpreted as Exploitation, Sincerity, and Institutional Trust. This factor solution was cross validated in a confirmation sample of 196 volunteers. Cosines between corresponding factors were high across samples and across sex. The similarity was pointed out between the present factor structure and the results of earlier studies with college students. Implications for possible refinement of the ITS were discussed. PMID- 16367568 TI - Measuring Machiavellianism with Mach V: a psychometric investigation. AB - This study re-examines the reliability of the Mach V Scale using two different scoring procedures: Christie's scoring system and an alternative scoring system by Rogers and Semin. Using commonly accepted standards, results for 123 business managers indicate that the Mach V Scale is not a reliable instrument. Regardless of the scoring Scheme or the method of assessment employed, reliability estimates found in this study are well below conventionally accepted criteria and are also well below reliability estimates previously reported in the literature. In addition, the investigation indicates that although the alternative scoring system appears to increase the precision of the Mach V Scale, the effect of this increase is negligible in hypothesis testing. PMID- 16367569 TI - Measures of autonomy. AB - Two studies examined the validity of 2 CPI autonomy scales developed by Kurtines (1973, 1978), In the first study, a criterion rating of autonomy based upon observer descriptions of adults (n = 314) was determined from the California Q set. This 10-item criterion was validated in a second study with college sophomores (n = 142) Using the criterion with a combined sample. of subjects, individuals high or low on autonomy were identified. Personality profiles of each group were determined from their responses to the CPI. Personality dimensions associated with ascendancy, self-assurance, interpersonal adequacy, and achievement orientation differentiated the two criterion groups. Correlation and multiple regression analyses indicated that Kurtines' first (1973) autonomy scale by comparison .predicted these personality dimensions Substantially better than his second (1978) scale, It was concluded, however, that neither scale adequately measured the global personality construct Of autonomy because (a) each scale assessed autonomous rule compliance to the exclusion of autonomous rule defiance, and (b)each scale measured only socially desirable aspects of autonomy. PMID- 16367574 TI - Who published what about whom? AB - Each study (n = 1,170) in the Journal of Personality Assessment and its predecessor (1968 through 1982) was reviewed to determine the affiliation of authors, personality measures commonly used, type of study methodologies employed, and characteristics of subjects studied. Generally, there were no large trends or shifts in any of these across the 15-year survey period. The Rorschach continues to be a common subject for study, as does the MMPI; authors have typically been from academic rather than applied settings; validity and normative studies have been most common; and the most frequently used subjects have been college students. Objective tests have increased in frequency in JPA articles, while projectives, except for the Rorschach, have declined; and the majority of research subjects have been drawn from nonpathological populations. Implications for editorial policies and reader interest are discussed. PMID- 16367575 TI - Shifts and constancies in Rorschach responses as a function of culture and language. AB - This study examined the relationship between Korean and American cultures and languages on Rorschach responses. The Rorschach Inkblot Test was individually administered to 20 Korean monolinguals, 20 American monolinguals, 20 Korean bilingual immigrants, and 10 American bilingual missionaries returned from Korea. The bilingual subjects took the test in Korean and in English. The results showed some shifts as well as some constancies in Rorschach responses as a function of culture. The results on the effect of language on Rorschach responses were less clear. PMID- 16367576 TI - Scale properties of the Holtzman inkblots. AB - Scaling techniques were presently applied to perceptions of inkblots, to empirically delineate the relationship between their stimulus properties and the nature of verbal associations elicited in projective testing. The Holtzman Inkblot Technique (HIT) was administered in group form to a relatively diverse group of college students. Subjects also individually rated the similarity of pairs of the HIT inkblots. Similarity judgments were analyzed via a multidimensional scaling (MDS) approach which recovered dimensions of variations among blots. The MDS procedures also captured variation across subjects in their utilization of blot stimulus properties. MDS solutions generally reflected differences among the blots along dimensions of physical characteristics of the blots. Differences in responsiveness of subjects to these characteristics appeared to reliably reflect meaningful substantive distinctions among subjects, many of which were not captured by traditional HIT variables. Implications were discussed in terms of further MDS applications and possible re-evaluation of HIT variables or procedures. PMID- 16367577 TI - Control orientation and the illusion of control. AB - Tested the hypothesis that individuals with an external locus of control are sensitive to the occurrence of noncontingency and will not display the illusion of control. Internals, who view outcomes as response dependent, should succumb to the illusion. One hundred thirty-eight students were asked to judge the amount of control they had over the onset of a noncontingent green light in one of two conditions: 25% green light onset and 75% green light onset. Both internals and externals demonstrated the illusion of control: they rated themselves as having more control when they received a green light on 75% of the trials, despite the fact that green light onset was response independent. These findings suggest that externals are doubly impaired. Their bias toward perceived response-outcome independence is given up when it would be most helpful, that is, when environmental cues suggest control in an objectively uncontrollable situation. PMID- 16367578 TI - Detecting (in)consistencies in personality: reconciling intuitions and empirical evidence. AB - After Mischel's (1968) critique of the traditional concept of personality, others have attempted to resolve the apparent discrepancy between intuitive (and theoretical) notions of consistency in the behavior patterns of individuals, and the available empirical evidence, much of which seems to suggest that the intuitive/theoretical notions are erroneous. Virtually all of these attempts have been grounded ultimately in some variation of the individual differences paradigm that has long dominated empirical personality research. In contrast, the present article suggests that this apparent discrepancy results from an attempt to reconcile essentially idiographic intuitions with aggregate empirical findings. Going beyond previous conceptual discussions of this point, the present article offers an empirical illustration of the problem, and suggests that the intuitions vs. empirical evidence discrepancy regarding questions of (in)consistencies in personality reflects the "incorrectness" of neither, but rather the fact that the intuitions and empirical evidence speak to fundamentally different questions. The implications of this fact for programmatic, theoretically oriented personality research are emphasized. PMID- 16367579 TI - Assessing variability in circumplex models of personality. AB - This paper describes a new methodology for assessing variability in circumplex models of personality. Leary's (1957) method for assessing such variability within his system of interpersonal diagnosis is discussed and critiqued. The authors then propose a new methodology, which is (a) consistent with assumptions underlying circumplex orderings of variables, and (b) generalizable to other circumplex models of personality and/or interpersonal communications. PMID- 16367580 TI - SACRAL: toward the meaning and measurement of friendliness. AB - Friendliness was regarded as a construct or dispositional tendency consisting of four components: Self-concept (S), the person's beliefs about self related to peer relationships; Accessibility (AC), giving behaviors involving attention and respect; Rewardingness (R), the giving of more tangible rewards, such as money and compliments; and Alienation (AL), personal beliefs about acceptance and the world as a friendly place. A 40-item questionnaire was developed to assess these components of friendliness. Studies of the questionnaire with college students and children supported its internal consistency, test-retest reliability, concurrent validity, construct validity, and discriminant validity. Although virtually all respondents reported themselves as friendly, high SACRAL scorers, when compared with low scorers, less frequently experience loneliness and in a variety of contexts claim to be more apt to engage in friendly behaviors. The impression that emerges of relatively low scorers on SACRAL is of people whose professed actions are somewhat inconsistent with their personal beliefs. PMID- 16367581 TI - Motives, values and moral judgments. AB - This study examined McClelland's (1981) hypothesis that operant and respondent measures of personality are orthogonal and assess different dimensions of personality structure. An operant measure of motives, a sentence completion test, and two respondent measures of cognitive schema variables, the Rokeach Value Survey and the Defining issues Test, were administered to 311 subjects. As predicted and in support of McClelland's hypothesis, 69% of the correlations between the respondent measures were significant at the alpha .05 level of probability versus 14% of the correlations between the operant and respondent measures. Further, within the domain of cognitive schema variables, it was theoretically possible to predict which values would correlate with different levels of moral judgment. The findings were discussed in terms. of their implications for person x situation models of social interaction and the prediction of criterion variables from typologies of personality. PMID- 16367584 TI - A meta-analysis of the reliability and validity of the Rorschach. AB - The results of a meta-analysis of Rorschach studies indicate that reliabilities in the order of .83 and higher and validity coefficients of .45 or .50 and higher can be expected for the Rorschach--when hypotheses supported by empirical or theoretical rationales are tested using reasonably powerful statistics. Three important determinants of variance accounted for in a variety of Rorschach scores were identified in 530 statistics from 39 papers published in the Journal of Personality Assessment from 1971 to 1980. The a priori theoretical or empirical evidence determining the likelihood of obtaining significant results, and the power of the statistic used to measure the results, as well as the interaction between the likelihood of results and the power of the statistic used, were all significant determinants of the proportion of variance accounted for in the Rorschach measures reported. PMID- 16367585 TI - Scoring the Rorschach: a constructive critique of Exner's Comprehensive Rorschach System. AB - John Exner's rationale for scoring the Rorschach and his decision to score "blend" responses by assigning equal weights are critically reviewed. The resultant exclusion of new responses made during the Inquiry of the Free Association responses is viewed as a major deficiency in the Comprehensive Rorschach System. A change to Klopfer's "Main-Additional" scoring method is recommended as a corrective. PMID- 16367586 TI - Card pull in projective testing. AB - Card pull, the tendency of the test stimuli to evoke or predispose certain cognitive and affective responses in the subject, is essential to the understanding and practice of projective testing. The basic instruction in projective testing employs the subjunctive mood to catalyze the dialectic between actuality and potentiality inherent in the projective stimulus. The subject is encouraged to play with the stimulus, to modify fact, to forsake the literal and the concrete, and to toy with the reality-stuff of the stimulus. Various conceptions of projective stimuli are reviewed and considered nondefinitive of projective testing. Projective testing is tentatively defined as a state of mind, a temporary but adaptive collusion shared between tester and subject which involves the former inviting the latter to modify a sample of reality. PMID- 16367587 TI - Locus of control and peer status: a scale for Children's Perceptions of Social Interactions. AB - A total of 985 third through sixth grade children participated in the development of a 48-item Locus of Control Scale for Children's Perceptions of Social Interactions (LOC-CPSI). Scores are based on the number of positive, negative, and total social reinforcers a child attributes to his or her own behavior (internal control). A social desirability subscale is included. Reliability coefficients were comparable to or greater than those of existing locus of control measures. Convergent validity was demonstrated by low, significant correlations with other locus of control measures. Internal scores correlated significantly with sociometric measures of popularity and predicted peer status better than other locus of control measures. Theoretical implications and potential clinical application are discussed. PMID- 16367588 TI - Body-image, personal space, and self-esteem: a field study. AB - In an earlier study, the author reported changes in body boundary perception and interaction distance resulting from the experimental manipulation of self-esteem. The present study attempted to replicate these laboratory findings in a field setting. The test battery included a self-esteem questionnaire and was administered to Army recruits before and after participation in a leadership training course. Results showed that while self-esteem had been modified predictably by performance, interaction distance was only partially affected, and the boundary indices remained unchanged. Results were discussed in relation to immediacy effects on the evaluation of competence, and their implications for field as compared to laboratory research. PMID- 16367589 TI - Sex-typing and cross-sex-typing in "androgynous" subjects. AB - Bem Sex-Role Inventory protocols from 848 subjects were categorized as androgynous or nonandrogynous according to each of three current scoring systems. Androgynous protocols by each scoring system were then examined for sex differences in trait item endorsements. The t-score method generated androgynous subjects who were cross-sex-typed, while androgynous subjects defined by the median split method tended to be same-sex-typed. Androgynous subjects by the intersect method presented the fewest number of sex differences. These method specific versions of androgyny appear linked to the use of summated Masculinity and Femininity subscales of the BSRI. PMID- 16367590 TI - CPQ personality factors of upper elementary gifted students. AB - The results of administering the Children's Personality Questionnaire (CPQ) to gifted students in the fourth, fifth, and sixth grades indicated that they had personality profiles similar to the norm group. Gifted males were more controlled, socially precise, self-disciplined, and compulsive than females, described as more casual and careless of social rules. Also, gifted females were more assertive and independent than males who had scores suggesting a more obedient, mild, or conforming personality. In comparing personality factors across grades, fourth graders had profiles significantly different from sixth graders. PMID- 16367599 TI - An error in citation of the Lb Scale of the MMPI. PMID- 16367591 TI - Cathectic orientation, goal setting and mood. AB - Investigated the reliability and validity of a new verbal cathexis test, based on objectively scored achievement data. Criteria were based on interviews about goal orientations and mood ratings, performed three times a day for a week. Goals were scored analogously with the cathexis test, viz. in the three dimensions hetero homo, active-passive, and positive-negative. Average mood scores and mood variabilities were estimated. The Oedipal and combined cathexis adjustment indices correlated with goal ratings. Moods correlated with cathectic balance indices. Strong correlations were found to exist between average moods and goal ratings in both active-passive and positive-negative dimensions, as well as sex differences in mood variability and Oedipal orientation as measured by the cathexis test. PMID- 16367600 TI - Artistic style and personality in creative painters. AB - The present study attempted to demonstrate a correspondence between artistic style and the personality of the creator. Personality style, defined as a cognitive-affective system of defenses and controls, was identified for 20 renowned painters through an analysis of their individual Rorschach protocols. Their creative work was classified independently by a jury of 4 persons (one art critic; two eminent painters and the junior author) into stylistic categories similar to those elaborated by the senior author for the Rorschach. The overall correspondence achieved was statistically significant (.72, p < .01), with individual correlations ranging from .75 to .86 (p < .01). PMID- 16367601 TI - A voice-operated method for administering the MMPI. AB - Paper/pencil psychological tests such as the Minnesota Multiphasic Personality Inventory (MMPI) are difficult to complete independently by patients with limited hand function. This paper describes a voice recognition system developed to allow patients with no hand function to take the MMPI independently. Profiles generated by this system are not statistically different from those generated by standard MMPI booklet format. Limitations and possible extensions of the system as it currently exists are discussed. PMID- 16367602 TI - The Cognitive Bias Questionnaire: further evidence. AB - Provided further evidence for the validity of the Cognitive Bias Questionnaire (Krantz & Hammen, 1979) as a measure of depressive cognitions. Forty depressed students completed the Cognitive Bias Questionnaire as well as general measures of depression and measures of depressed mood. In addition, independent raters counted frequency of negative and positive self-statements generated during a tape recorded self-description, and rated Concurrent depressive tone, The results replicate previous findings of convergent and discriminant validity for the Cognitive Bias Questionnaire. Furthermore, depressed-distorted responses and nondepressed-nondistorted responses were significantly correlated with independently assessed depressive verbal behaviors. PMID- 16367603 TI - Personality attributes of ultramarathoners. AB - The Self-Motivation Inventory (SMI) and four subscales from the Philosophies of Human Nature were given to 50 male ultramarathoners who were matched for age and sex with runners who had never raced any distance longer than 10 miles, and with a group of nonrunners. No significant personality differences were obtained among the three groups. Experienced ultramarathoners were Significantly. more skeptical about the goodness of human nature than inexperienced ones. PMID- 16367611 TI - The concurrent validity of child Rorschach interpretations. AB - Rorschach records were obtained by 10 examiners from 59 preschool children. These records were reliably scored and interpreted. Interpretation consisted of preparing a list of Rorschach concepts for each child. An additional number of Rorschach concepts of equal frequency in this sample were added to each child's list of concepts. Half of the concepts on each child's list were checked for agreement by school personnel. The children were described with consistency across judges and over time by the same judges. The school personnel were able to differentiate concepts that described each child from nonrelevant concepts. The Rorschach concepts provided data that was validated by teacher perception of these children. This methodology for Rorschach validation makes use of interpretive concepts derived directly from data rather than from reports. PMID- 16367612 TI - Male-oriented vs. balanced-as-to-sex thematic apperception tests. AB - Measures of need Achievement, need Affiliation, and need Power were obtained for 124 working managers (62 female, 62 male) using both McClelland's traditional male-oriented thematic apperception test (TAT) and a set of picture cues balanced as-to-sex. Two-way analysis of variance was performed in a 2 x 2 factorial design (sex by version of TAT). F tests were conducted to test significance at the .05 level. Results indicate both sets are interchangeable in determining need Achievement and need Power distinctions between managerial women and men, Absolute scores are also comparable. For need Affiliation, however, motivation scores are not comparable and interchangeability is clouded, suggesting cue modification may be needed. PMID- 16367613 TI - Fear of success: the construction and validation of a measuring instrument. AB - This study constructed an instrument which identifies people who fear success (FOS) in academic areas and conducted a laboratory study which tested the instrument's predictive validity. The theoretical formulation of the FOS phenomenon leads to the expectation that individuals high in the fear of success would demonstrate the following characteristics: low self-esteem, a preoccupation with the evaluative aspects of situations, a competitive orientation, repudiation of competence, and self-sabotage at the approach or attainment of success. The fear of success questionnaire is an 83-item scale with a reliability of .90. In a 2 x 2 factorial design, subjects identified by the FOS questionnaire were given either Success or nonsuccess feedback after completing the first of two equivalent reading tests. The results of the laboratory experiment indicated that there is strong clear evidence of self-sabotage under conditions of success among subjects who scored high on the FOS questionnaire. PMID- 16367614 TI - Variations of the Bender-Gestalt Test: implications for training and practice. AB - Although the Bender-Gestalt Visual Motor Test has been a popular and influential clinical assessment instrument for the last half century, authoritative opinion and a substantial research literature are often inconsistent or contradictory. This paper documents variations in designs, administration procedures, forms, and applications to particular populations. A careful review of scoring systems indicates the current status of objective approaches as contrasted with interpretation that is global, intuitive, and subjective. Some implications of these variations and interpretive procedures for training and assessment practice are discussed. The major conclusions of the review are summarized in seven points. PMID- 16367615 TI - The Common Belief Inventory for Students: a measure of rationality in children. AB - The Common Belief Inventory for Students (CBIS) was developed to create a measure of rationality in children. The CBIS was formulated within Ellis's conceptual framework of Rational Behavior Training. It was constructed to yield a total score as well as a score for each of Ellis's 11 irrational ideas. A pretest/post test design was employed over experimental and control groups (n = 1,226), Subjects in the experimental group were exposed to one hour of Rational Behavior Training each week for six weeks. Results indicated that the items were reliable indicators of the total test score and the belief scores. Belief components were reliable indicators of the total test score. Total test reliability estimates were adequate. The CBIS also appeared sensitive to the use of RBT methods to enhance rational thinking. PMID- 16367616 TI - A three dimensional test for White's effectance motive. AB - A scale was developed based on Robert W. White's construct of effectance motivation. Effectance was conceptualized as involving three dimensions--Effect on Self, Effect on Objects, and Effect on People. It was found that reliabilities for the three subscales were good. Validity was determined using three methods: criterion groups, multitrait-multimethod matrix, and convergent and discriminant validity. Validity was supported under all three methods. PMID- 16367617 TI - Personality structure of Mexican Americans using the Comrey Personality Scales. AB - The Comrey Personality Scales (CPS) were used to examine the personality structure of 176 Mexican American college students. A majority of the eight factors maintained substantial loadings and corresponded closely with Comrey's normative sample as well as with Brazilian and New Zealand samples. Two factors, Conformity and Orderliness, showed the weakest fit across all four groups. Overall, the CPS shows adequate measurement properties for "normal" personality assessment among Mexican Americans and is preferable to instruments such as the Minnesota Multiphasic Personality Inventory (MMPI). However, the external validity of the CPS with clinical populations must first be determined prior to its use to assess psychopathology among Mexican Americans. PMID- 16367620 TI - Sex differences in primary process thinking and flexibility in problem-solving in children. AB - The relationship between primary process thinking and flexibility in problem solving was investigated. Specifically, it was hypothesized from psychodynamic theory that primary process integration on the Rorschach (Holt's scoring system) would be positively related to the capacity to shift problem-solving strategies on the Luchins' water-jar test. The Rorschach and water-jar test were individually administered to 47 third grade children. The hypothesis was confirmed for boys in that the Adaptive Regression score on the Rorschach was significantly related to performance on Luchins' water-jar test (r = .64, df = 16, p < .01). This relationship remained significant when IQ was partialed out (r = .59, df = 16, p < .01). There was no relationship between primary process integration and flexibility in problem solving for girls (r = .02, df = 26, NS). These results and sex differences were replicated with a second sample of children. Consistent sex differences also were found in the amount of primary process material expressed. Girls showed a significantly smaller percentage of primary process material than boys. Primary process integration was also related to Iowa test achievement scores, independent of IQ. This finding is consistent with previous studies. PMID- 16367621 TI - Primary process patterning in college students' inkblot responses. AB - Holzman Inkblots were administered under standard instructions and under instructions to respond imaginatively. Protocols were scored according to Holt's system for scoring primary process. Subcategories of primary process proved to be reliable and to differ in prominence. A stable pattern of primary process, characterizing college students' inkblot responses, appears to represent students' intuition of what it means to be imaginative. PMID- 16367622 TI - Intimacy motivation and psychosocial adjustment: a longitudinal study. AB - Longitudinal data of 57 middle-aged men from the Grant Study Adult Development were analyzed in terms of nine indices of psychosocial adjustment and four social motives: achievement, power, affiliation, and intimacy motivation. The Thematic Apperception Test (TAT), administered in 1950-52, was scored for the four motives. Psychosocial adjustment, determined by ratings made primarily with reference to life history data gathered between 1950 and 1967, was comprised of scores on income level, occupational promotion, occupational enjoyment, days of sick leave, marital enjoyment, regular vacations, pastimes with friends, drug or alcohol misuse, and psychiatric visits. High intimacy motivation at age 30 was significantly associated with better adjustment 17 years later. The results are discussed in terms of contemporary theories of psychosocial adaptation. PMID- 16367623 TI - Human figure drawing size as a measure of self-esteem. AB - The relationship between self-esteem and human figure drawing size was investigated. Seventy-six adolescents completed the Rosenberg Self-Esteem Scale and drew the human figure. For males only. there was a significant linear relationship between self-esteem and width of human figure drawing and a significant curvilinear relationship between self-esteem and height and self esteem and area of human figure drawing. The latter findings lend support to a hypothesis regarding the possibility among low self-esteem subjects of a bimodal distribution with respect to size. PMID- 16367624 TI - Sex-role orientation and psychological adjustment: comparison of MMPI profiles among college women and housewives. AB - Androgynous, masculine, feminine, and undifferentiated women, as determined by the Bern Sex Role Inventory, were compared on the three validity and ten clinical scales of the MMPI. Two samples of women were included: Female college students and a group of upper middle-class suburban housewives. No significant differences emerged in the housewife sample. Among college women, androgynous and masculine women did not differ in adjustment, and both groups were better adjusted (i.e., less depressed, anxious, tense, and less socially introverted) than the undifferentiated women. A cross-validation study with a comparable student sample essentially replicated the results of the first study. Rather than supporting Bem's (1974) hypothesis of better psychological adjustment in androgynous persons, results were interpreted as providing support for the notion that masculinity is associated with equally good adjustment as androgyny in college women. PMID- 16367625 TI - Development of the Revised Sex-Role Behavior Scale. AB - This study describes the construction of the Revised Sex-Role Behavior Scale (SRBS-2), a 240-item self-report inventory comprised of male-valued, female valued, and sex-specific items in four interest/behavior areas: Recreational and leisure activities, vocational preferences, social interaction, and marital behaviors. Two-hundred sixteen items were administered to 520 undergraduate males and females under three instructional sets. One-third of the subjects compared the typical young adult man and woman on the items, one-third rated the desirability of the items for men and women, and the remaining subjects rated themselves. Subjects in this latter condition also rated the 160 items from the SRBS-1. Typical and desirability ratings were used to select items to add to the SRBS-1 male-valued (more typical of men but desirable for both sexes), female valued, and sex-specific (more typical of one sex and desirable only for that sex) scales. The resulting overall and area subscales had adequate internal consistency. Intercorrelations among the area subscales showed a fair degree of consistency across behavior areas, suggesting that global sex-role behavior assessments may be obtained with the overall SRBS-2 scales but that more detailed and accurate assessments require the use of separate area subscale scores. PMID- 16367626 TI - A method for measuring "primitive" and "advanced" elements in pleasures and aversions. AB - Subjects were asked to list pleasures and aversions actually experienced, and pleasures and aversions based on their "innermost fantasies." A 7-point rating scale was developed to rate "primitive" and "advanced" elements in these pleasure aversion responses. The "primitive" pole of the scale was keyed with the phrase "gut-level experiencing" and the "advanced" pole with "detached objective thinking." Reality Pleasure, Fantasy Pleasure, Reality Aversive, Fantasy Aversive, and Total scores were derived by having trained raters rate each subject's response with the 7-point rating scale. In Study 1, 25 females and 25 males provided pleasure-aversion responses and the data analyzed in a mixed 2 (male-female) x 2 (reality-fantasy) x 2 (pleasure-aversive) ANOVA. The most important finding was a highly significant reality-fantasy x pleasure-aversive interaction: subjects tended to "progress" (give more advanced responses) going from reality to fantasy in their pleasure responses, while exactly the opposite occurred with the aversive responses. Also, as predicted, reality aversive responses tended to be more "advanced" than reality pleasure responses. Study 2 was similar in design, but several additional personality measures were administered to assess the construct validity of the pleasure-aversion scores. As in Study l, the reality-fantasy x pleasure-aversive interaction was highly significant and again reflected a pattern of "progression" going from reality to fantasy for pleasure responses and "regression" for aversive responses. The correlation analysis also produced several interesting findings with sex serving as a major moderator variable. PMID- 16367630 TI - Body-image, personal space and self-esteem: preliminary evidence for "focusing" effects. AB - A repeated-measures design was used to investigate the effects of a self-esteem manipulation on measures of body-image and personal space. Significant interaction effects were obtained for penetration, body-cathexis, and simulated interaction distance, and the results confirmed the prediction that subjects whose self-esteem had been enhanced would obtain significantly higher body cathexis, and significantly lower penetration and interaction distance scores than those whose self-esteem had been reduced. Although the interaction effect was nonsignificant, similar trends were observed for barrier. Figure drawing and spatial orientation appeared to be unrelated to self-perception. Factor analysis showed that the tests were unrelated, but the self-esteem manipulation had a "focusing" effect on boundary and interaction distance. Overall, the results provide experimental support for the defensive distancing hypothesis. PMID- 16367631 TI - Some reflections on "MMPI short forms: a literature review". AB - Stevens and Reilley (1980) have suggested that recent changes in the methodological approaches for developing short forms of the MMPI make them an expanding and fruitful area of research. Since Stevens and Reilley did not address several important issues in the development and use of Short forms, these issues are reviewed and suggestions are presented for future research with short forms. PMID- 16367632 TI - Factor analytic study of the Piers-Harris Children's Self-Concept Scale. AB - A factor analysis of the Piers-Harris Self-Concept Scale was conducted with a biracial u sample of children over a wide age range. The Scale was completed by 348 10-17 year old students in a rural community. Seven factors of 10 had sufficient item loadings to be interpretable. Six matched those found in the manual and an additional factor was found relating to Aggression. Race, sex, and age relationships are also reported for these seven factors. PMID- 16367633 TI - The assessment of social intimacy. AB - Research exploring the psychological importance of closeness with others has been hampered by the absence of a reliable and valid measure of this variable. The development of the Miller Social Intimacy Scale (MSIS), a 17-item measure of the maximum level of intimacy currently experienced, is presented. Evidence for internal consistency and test-retest reliability as well as for convergent, discriminant and construct validity is discussed in the context of the need for further scientific exploration of this important phenomenon. PMID- 16367634 TI - Sex-roles of ideal opposite sexed persons for college males and females. AB - Relationships between sex-roles as measured by the Bem Sex-Role Inventory (BSRI) and the ideal sex-role of persons of the opposite sex as measured by a modified BSRI were investigated for 169 college males and 204 college females. The distributions of sex-roles were different for men and women, as were the distributions of ideal sex-roles of opposite sexed persons. Androgynous (A), feminine (F) and masculine (M) men identified F women as ideal, while undifferentiated (U) men identified U women as ideal. M women preferred M men, and U women preferred U men. Effects of ideal sex-role of the two sexes on sexual attraction were discussed. PMID- 16367635 TI - The development of a Money Attitude Scale. AB - On the basis of the clinical and theoretical literature, 62 items in the domains of security, retention, and power-prestige were generated. Factor analysis yielded five factors: power-prestige, retention-time, distrust, quality, and anxiety. A 29-item Money Attitude Scale (MAS) was developed on the basis of four of these factors. Reliability measures, as well as personality and psychopathological correlations, were determined and discussed. PMID- 16367636 TI - Changes in identification during adolescence: a personal construct theory approach. AB - The Role Construct Repertory Grid was administered individually to groups of eight subjects representing the early, mid and late adolescence age ranges. The Mid-Adolescence Group showed significantly greater conceptual distances between ideal-self and actual-self than either of the other groups. Differences between the age groups on self-integration plots and a measure of self-isolation were also observed. Evidence was found of decreasing identification with parents as a function of age. PMID- 16367637 TI - A self-concept measure for preschool and early primary grade children. AB - The article describes the development of the Preschool and Primary Self-Concept scale. Subjects of the standardization sample were 109 boys and 127 girls, ages 4 to 9, enrolled in preschool and primary educational programs. Boys described themselves as significantly stronger O and bigger than girls. Girls described themselves as significantly more go d than boys. Older children described themselves as significantly more active than younger children. Factor analysis of the scale revealed an evaluative and gender dimension. Subjects of the validation study were 46 first graders. Satisfactory convergent and discriminant validity were demonstrated with the multitrait-multimethod technique. PMID- 16367638 TI - Personality description from the WAIS: a comparison of systems. AB - The present study compares an atheoretical, actuarial method for obtaining personality and behavioral descriptions from WAIS profile patterns to the Personality Assessment System--the theoretical system developed by J. Gittinger for the same purpose. Interest was focused on both the determination of group membership by the two systems, and on the resultant personality descriptions. The PAS theory based sorting systems did identify the target group of the actuarial approach well, and the personality descriptors were found to have distinct resemblance, though certainly not complete overlap. PMID- 16367643 TI - Sex and gender bias in self-report personality disorder inventories: item analysis of the MCMI-II, MMPI, and PDQ-R. AB - There has been considerable controversy and research regarding sex bias in the diagnosis of personality disorders, but little has involved self-report inventories. Thus this study investigated items from the Millon Clinical Multiaxial Inventory-II (Millon, 1987), the Minnesota Multiphasic Personality Inventory (Morey, Waugh, & Blashfield, 1985), and the Personality Diagnostic Questionnaire-Revised (Hyler & Rieder, 1987). Subjects (N = 189) completed the Histrionic, Dependent, Antisocial, and Narcissistic scales from these inventories, along with the Bem Sex Role Inventory (Bem, 1974) and the Symptom Checklist-90-Revised (Derogatis, 1977). Items were considered to evidence sex or gender bias if they (a) failed to correlate with dysfunction and (b) exhibited sex or gender role differences. At least 13 items evidenced sex bias (76 items using a more liberal threshold). The majority were from Narcissistic scales; few Histrionic items evidenced sex or gender bias. Implications with respect to sex bias assessment and item construction are discussed. PMID- 16367644 TI - Diagnosing personality disorders: an examination of the MMPI-2 and MCMI-II. AB - Personality disorders are highly prevalent in clinical populations and affect outcomes across all forms of intervention. This investigation examined the diagnostic efficiency of two widely used, self-report measures of personality disorder (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989; MCMI-II; Millon, 1987), as compared to a structured interview (SCID-II; Spitzer et al., 1987) diagnosis. The measures were administered to 150 residential and outpatient volunteer subjects. Persons with primary organic or psychotic-spectrum disorders were excluded from participation. Results were variable across disorders measured, with low to moderate levels of diagnostic agreement observed. The MCMI II appears to be a more sensitive measure, whereas the MMPI-2 is more specific. The two self-report measures demonstrated greater convergence with each other than with the interview measure. Both the MMPI-2 and MCMI-II were more accurate at identifying the absence of a given disorder. Although overall diagnostic powers exist at acceptable levels. the results suggest that diagnoses generated by self-report versus interview are not interchangeable. PMID- 16367645 TI - How (not) to evaluate the comparability of MMPI and MMPI-2 profile configurations: a reply to Humphrey and Dahlstrom. AB - Humphrey and Dahlstrom (1995) presented a study on the comparability of MMPI/MMPI 2 (Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) profiles in which they concluded that "the bases for clinical interpretation derived from the MMPI and MMPI-2 profiles were sufficiently at variance to require different conclusions" [sic] (p. 2). In this brief critique, we identify procedural and data-analytical deficiencies that invalidate Humphrey and Dahlstrom's argument. Their blanket recommendation based on this argument, namely, that clinicians routinely plot both MMPI and MMPI-2 profiles, is unwarranted. PMID- 16367646 TI - A 'big five' scoring system for the item pool of the adjective check list. AB - The item pool of the Adjective Check List (ACL; Gough & Heilbrun, 1980) is widely used as a means of capturing the personal characteristics associated with various target groups (e.g., women vs. men, young adults vs. old adults). The purpose of this research was to develop a system for scoring the ACL items in terms of the five-factor model of personality. In Study 1, five groups of introductory psychology students served as judges, with each group of approximately 100 persons rating the 300 ACL items for one of the five factors. The ratings of each factor were highly reliable. When corrected for favorability, the intercorrelations among the five factors were quite low, as expected, except for the positive correlation of Openness and Extraversion. Good convergence was found between our ratings and the indicative and counterindicative items identified by John's (1989) graduate student judges. In Study 2, convergent validity was demonstrated between the five-factor scores obtained from self-descriptive ACLs and corresponding factor scores obtained from Costa and McCrae's NEO-PI-R and NEO FFI instruments (Costa & McCrae, 1992). Data from earlier cross-cultural studies of gender and age stereotypes were rescored using the new ACL-FF system to illustrate its potential utility as a research tool. PMID- 16367647 TI - A note on the assessment of idealization. AB - In a series of studies using the Lerner Defense Scale (H. Lerner & P. Lerner, 1980) for assessing primitive defenses, inconsistent and at times perplexing findings have been reported regarding the defense of idealization. A major study examining personality variables, including defense, in a nonclinical population ballet students-and comparing them to those of patients suffering eating disorders, yielded findings related to the defense of idealization and its assessment. Herein, the results of that study are reviewed and then discussed in light of recent psychoanalytic formulations regarding idealization. PMID- 16367648 TI - Depressive experiences questionnaire: an empirical exploration of the underlying theory. AB - This article addresses the theoretical conceptualization of depressive typology proposed by Blatt (1974) by analyzing selected items on the Depressive Experiences Questionnaire (DEQ) in a 21-item modified version. Items were selected by judges and by factor loading criteria to be most theoretically characteristic of the dimensions they are meant to represent: anaclitic and introjective depression. Two independent samples, a female inpatient sample and a female college sample, were used. Principal components analysis of these 21 items revealed only 5 anaclitic items and 7 introjective items that loaded on their respective factors for both samples. Inspection of these items suggests that the anaclitic depressive experience is characterized by discomfort with interpersonal separation, whereas the introjective experience is characterized by negative self evaluation with respect to self-imposed standards. Other putative aspects of these two depressive dimensions were not supported by this study; particularly, guilt and self-blame were not associated with introjective depression. PMID- 16367649 TI - Confirmatory factor analyses of seven locus of control measures. AB - Confirmatory factor analyses with congeneric models were used to examine whether seven common locus of control instruments were measuring the same single construct. The subjects were 191 adolescents, and the analyses were based on the intercorrelations among the scales as reported by Furnham (1987). In addition to the one-factor congeneric model, other first- and second-order factor models were posited to reflect the effects due to target subject (adult vs. child), dimensionality (uni- vs. multidimension), and scaling method (yes/no, forced choice, 7-point). A recently developed strategy in multitrait-multimethod analyses was also adopted in the analyses. In general, results of this study did not substantiate the claim that all scales were measuring the same construct. Even among the more strongly correlated unidimensional measures for children, the support for a one-factor model was still rather weak. Furthermore, results did not justify the categorization of the measures by their target subject, dimensionality, and scaling method. PMID- 16367650 TI - The Perception of Teasing Scale (POTS): a revision and extension of the Physical Appearance Related Teasing Scale (PARTS). AB - This article describes the development and validation of a revised measure for the assessment of history of being teased about physical appearance. Study 1 involved the preliminary psychometric evaluation of the questionnaire on a sample of 227 college women, Two factors emerged: Weight-Related Teasing (WT) and Teasing About Abilities/Competencies (Competency Teasing; CT). The integrity of the factor structure of these two scales was established in Study 2 with a sample of 87 college women. Internal consistency ratios in this sample were also found to be acceptable. In Study 3, 92 college women were administered measures of body image, eating disturbance, and self-esteem to test for convergence with the Perception of Teasing Scale. Subjects also rated teasing items for frequency and effect (e.g., how upset they were by the teasing). Two-week test-retest reliabilities for these measures were acceptable. Weight-Related Teasing correlated to a greater degree with other measures than Competency Teasing. Regression analyses revealed the importance of a frequency versus effect dimension. Findings are discussed in light of recent research on developmental factors in body image and eating disturbance. PMID- 16367651 TI - The relationship of the Yale-Brown Obsessive Compulsive Scale (YBOCS) to other measures of obsessive compulsive symptoms in a nonclinical population. AB - The Yale-Brown Obsessive Compulsive Scale (YBOCS; Goodman, Price, Rasmussen, & Mazure, 1989a) is an interview-based rating scale measuring severity of obsessive compulsive disorder (OCD). Because it is independent of the number and type of OCD symptoms and minimizes confounding with other types of symptoms, it has become the "gold standard" for assessing the outcome of behavioral and pharmacological treatments. This study was designed to further validate the YBOCS in relation to self-report measures of obsessive compulsive phenomena in a nonclinical population. Among a group of 45 female college students, the three primary YBOCS measures (obsessions, compulsions, and total score) were internally consistent and correlated moderately to strongly with self-report measures of obsessive compulsive phenomena that have been used in previous research. The compulsive subscale of the YBOCS showed the lowest correlation with self-report measures sharing only 25% of the common variance. This measure is appropriate for use with nonclinical samples and may prove superior to other instruments for detecting the presence and severity of obsessive and compulsive symptoms. PMID- 16367652 TI - Maria Rickers-Ovsiankina: a Russian expatriate in America. A review of Rorschach Psychology. AB - This article is a review and reassessment of Rorschach Psychology, edited by Maria Rickers-Ovsiankina. The various chapters in the book are written with an attempt to tie Rorschach conceptualization to academic psychology and to various theoretical approaches. A wide range of theories are considered, including a gestalt psychology approach, an experiential-phenomenological approach, and several psychodynamic approaches (e.g., Freudian and Jungian theory). In addition, information is included about Rickers-Ovsiankina's early life and background, and the contents of many of the individual chapters are described and evaluated. PMID- 16367656 TI - Detection of faking on the MMPI-2: differentiation among faking-bad, denial, and claiming extreme virtue. AB - The purpose of this study was (a) to evaluate the ability of the existing validity indices of the Minnesota Multiphasic Personality Inventory (MMPI-2) in detecting faking. bad and faking-good profiles, (b) to differentiate between two strategies for faking-good (denial and claiming extreme virtue), (c) to determine the effectiveness of the new MMPI-2 validity scale, the S scale, in detecting people's tendency to present themselves in a superlative manner, and (d) to explore the response strategy under conditions in which individuals we known to be distorting their responses to the MMPI-2 Participants were 167 undergraduate, college students who were administered the test under different conditions, and 50 hospitalized, psychiatric patients. Mean profile configurations and optimal cutoff scores obtained in this study were similar to those reported in previous studies. Accurate identification of faked profiles was achieved. The new S scale and its subscales, especially S2 (Serenity), S3 (Contentment With Life), and S5 (Denial of Moral Flaws), showed particular promise in detecting faking-good (denial and claiming extreme virtue) profiles, Participant's debriefing provided valuable information about the participants' understanding of the instructions and their deception strategies. The attempts to differentiate between two strategies for faking-good, denial and claiming extreme virtue, were not successful, However, these results should not be taken to indicate that the particular strategies assessed in this study cannot be differentiated. Methodological issues reviewed suggest that further research might yield more understanding of the nature of any deception efforts made by respondents. In addition, future, research is needed to discover if the findings from this study generalize to other clinical settings and populations. PMID- 16367657 TI - Detection of a Cry for Help on the MMPI-2: an analog investigation. AB - We compared Minnesota Multiphasic Personality Inventory (MMPI-2) results from 2 groups of mildly to moderately disturbed psychiatric outpatients (N=60) answering under either Standard or Cry for Help instructions. Results from previously completed intake MMPI-2 protocols were obtained for each participant. The two groups were comparable on intake MMPI-2 variables and demographic characteristics. Analysis of feedback data from the second, experimental observation indicated that the two groups had equivalent self-estimates of understanding of their role and success in simulating their role. Significant differences were found between the two groups on their experimental MMPI-2 protocols. The Cry for Help group had significantly higher scores on F, F-K, Fb, Ds2, and Fp scales as well as significantly lower scorns on the K scale. The Cry for Help group also had significantly higher scores on all clinical scales with the exception of 5, which was not tested. Cutting scores derived by Rogers, Sewell, and Ustad (1995) for the detection of a Cry for Help in outpatients were cross-validated with fairly positive results. Although these results are promising, particularly in light of the shrinkage expected on cross-validation, further research is needed in this area. PMID- 16367658 TI - Levels of relatedness within the dependency factor of the depressive experiences questionnaire for adolescents. PMID- 16367659 TI - Five scales in search of a construct: exploring continued attachment to parents in college students. AB - The multitude of measures and differences across self-report indices of continued parental attachment raise questions regarding the validity and meaning of these scales. The purpose of this study was to examine the convergent and construct validity of 5 measures of continued parental attachment. Five attachment scales and 6 personality scales were administered to 216 undergraduate students. Factor analyses and correlational analyses indicate that the 5 attachment measures differentiate healthy from pathological bonding with parents, the construct being assessed has multiple dimensions, and scores on these attachment measures are correlated with personify variables as would be expected. These attachment scales appear to assess a construct that is related to attachment, although they may be more illustrative of the general affective quality of relationships. Subsequent assessment efforts should employ more specific ratings and measure concrete behavioral manifestations of attachment to most effectively examine the construct. PMID- 16367660 TI - Development and effects of an MMPI--A K-correction procedure. AB - Although the results of research on the effectiveness of the K-correction factor have been inconclusive, this procedure has been widely used with adult respondents to correct for defensiveness or underreporting of symptomatology on the Minnesota Multiphasic Personality Inventory. Although the K-correction procedure was incorporated into the Minnesota Multiphasic Personality Inventory 2, the Minnesota Multiphasic Personality Inventory-Adolescent (MMPI-A) was developed based exclusively on the use of non-K-corrected T scores. This study derived age-appropriate K-weights for the MMPI-A to determine the degree to which the use of this procedure could improve test accuracy in the classification of participants into normal and clinical groups. Discriminant function analyses were performed to determine the K-weight that, when combined with basic scale raw score values, optimally predicted normal versus clinical status for each of the eight basic clinical scales. Hit rate analyses were utilised to assess the degree to which K-corrected T scores resulted in improvements in classification accuracy in contrast to standard MMPI-A non K-corrected norms. Results indicate that the adoption of K-correction procedure for the MMPI-A does not result in systematic improvements in test accuracy and the current findings do not support the clinical use of a K-correction factor in interpreting MMPI-A protocols. PMID- 16367661 TI - Contributions of cognitive science to the Rorschach Technique: cognitive and neuropsychological correlates of the response process. AB - This article presents an understanding of the Rorschach Technique in terms of emergent models from cognitive science. We propose a linkage between cognitive psychology and neuropsychology in understanding the operations that underlie the Rorschach response process. Contemporary information processing models are described. The Rorschach Technique is conceived of as a complex process involving all areas of the cerebral hemispheres, encompassing various aspects of visual attention and perception, object recognition, associative memory, language production, and executive functioning. Exner's model of the response process is delineated, including both Association and Inquiry phases, in terms of requisite underlying neuropsychological abilities and anatomical substrates. The question of the Rorschach Technique's status, utility, and potential as a neuropsychological assessment, tool is discussed. Understanding the Rorschach in terms of contemporary cognitive psychological and neuropsychological models heightens appreciation of the technique's complexity and provides a heuristic and conceptual foundation for empirical research. PMID- 16367666 TI - Current perspectives on psychoanalytic Rorschach assessment. AB - In this article, I present as clear and as updated an account as possible of several of the essential features that comprise a psychoanalytic approach to the Rorschach. Specific attention is accorded the assessment frame, the language of assessment, sources of assessment information, and differing approaches to the interpretation of Rorschach data. The article comes out of a Rorschach tradition begun by Rapaport, extended by Schafer and Mayman, and presently being updated by Leichtman and Smith. PMID- 16367668 TI - The nature of the Rorschach task. AB - This article offers a critique of traditional theories of the Rorschach as a perceptual task, proposes an alternative conception of the task as one of visual representation, and demonstrates the manner in which the latter theory provides a superior explanation of the distinctive quality of the Rorschach stimuli, the ways in which participants and examiners understand the test, the assumptions underlying Rorschach scoring, and the manner in which young children master the test. PMID- 16367669 TI - The interpretive process in Rorschach testing. AB - In this article, two complimentary and interwoven approaches to Rorschach interpretation are described. The first, a more structural approach, emphasizes test responses, the response process, and test rationales. Theorists with this emphasis focus primarily on the structural features of a test record?the scores and their interrelations. Interpretations drawn from the structural features of a record themselves tend to have a structural emphasis. A second approach, referred to as experiential, is more subjective and phenomenological. Based on the examiner's empathy, this approach is applied to other aspects of a Rorschach protocol such as content of responses and the testee's more spontaneous and off handed comments. Each of these interpretive approaches must be given its due and integrated with interpretations drawn from the other. PMID- 16367671 TI - MMPI-Rorschach interrelationships: proposed criteria for evaluating explanatory models. AB - An extensive empirical literature, spanning 50 years and 45 published investigations, leads to the conclusion that the Rorschach and the Minnesota Multiphasic Personality Inventory (MMPI) bear little or no meaningful relationship to each other. An inherent tension exists between these empirical findings and the widespread clinical practice of integrating MMPI and Rorschach results. At least three perspectives have been advanced concerning the relationship between these two instruments. One view holds that variables from the two measures will demonstrate significant patterns of convergence only in carefully designed research studies that involve specific, well-constructed and theoretically derived predictions. A second view postulates that although variables from the Rorschach and MMPI do not bear a high intercorrelation with each other, these variables may be combined to contribute significantly to the prediction of outcome variance. In this view, the combined use of the two instruments yields clinically useful increases in incremental validity not achievable by the use of either instrument in isolation. A third approach is reflected in the proposition that although the MMPI and Rorschach do not produce significant interrelationships under general conditions, meaningful relationships may be yielded under specific psychometric conditions, for example, under conditions in which the response styles are consistently displayed across those two instruments. This article will review research evidence related to each of these perspectives, with particular emphasis on the importance of clearly specifying the criteria to be applied in evaluating these models. The concepts of parsimony and heuristic value are proposed for these evaluation purposes. PMID- 16367672 TI - Remarks on method for MMPI-Rorschach studies. AB - An impressionistic list of methodological considerations that may be of consequence in Minnesota Multiphasic Personality Inventory (MMPI)-Rorschach studies is presented. These include the potential effects of the multidimensionality of the basic clinical MMPI scales in comparison studies, issues of deception and response style in such studies, and the rote of theory in guiding the selection of variables in the search for MMPI-Rorschach correspondences. The recently established necessity of controls for the total number of Rorschach responses provides the basis for suggestions for further controls in MMPI-Rorschach studies, including those for level of distress and for the minimization/exaggeration of psychopathology. An effort is made to contrast these two instruments in terms borrowed from information theory as a way of illuminating their complementary aspects, and suggesting means by which each may contribute to an enhanced understanding of the other. PMID- 16367673 TI - Exploring MMPI-Rorschach relationships. AB - Limitations exist in the current literature examining Minnesota Multiphasic Personality Inventory (MMPI)-Rorschach interrelationships. In this article I attempt to identify several issues to be addressed by researchers in this area. These include restricting conclusions about MMPI-Rorschach relationships to research using the Comprehensive System; considering the relative reliability and validity of these tests; comparing the predictive power of the Rorschach and MMPI; examining the effects extreme outliers have on statistical analyses; and determining the appropriate level of analysis for research, the level of individual test scores, or psychological constructs. The implications of these issues for research design, statistical analysis, and applied clinical work are discussed. PMID- 16367674 TI - Assessment feedback integrating MMPI-2 and Rorschach findings. AB - Many clinicians are committed to giving feedback to clients about assessment results, but puzzle over how to integrate Minnesota Multiphasic Personality Inventory-2 (MMPI-2) and Rorschach findings when talking to clients. When the two tests agree, findings may be combined and assessors may use the language of clients' Rorschach percepts to frame findings from the MMPI-2 or the Structural Summary. When the Rorschach shows more disturbance than the MMPI-2, assessors may discuss "levels of personality," praise clients for their usual coping mechanisms, and raise the possibility of underlying problems. When the Rorschach depicts less psychopathology than the MMPI-2, clinicians may talk with Clients about factors influencing them to present as needing help. Alternatively, assessors may hypothesize that clients constrict emotionally in interpersonal, unstructured situations. PMID- 16367675 TI - The Rorschach and MMPI: toward a more scientifically differentiated understanding of cross-method assessment. AB - Reasons for Minnesota Multiphasic Personality Inventory (MMPI) and Rorschach disagreement at the nomothetic level are explored. Building on an understanding of measurement distinctions from other sciences, it is proposed that the Rorschach and MMPI procedures are differentially sensitive to unique manifestations of personality. By necessity, each method is then also recognized as having a limited scope of effectiveness, such that neither can provide a complete picture of personality in its full complexity. Drawing on the more extensive self-report literature, the idiosyncracies and limited scope of the self-report method are documented. Finally, an ideographically rooted, cross method, configural model is proposed for validation research in personality assessment. Several examples consistent with this approach are drawn from the assessment literature and discussed. PMID- 16367676 TI - Data and issues to consider in reconciling self-report and the Rorschach. AB - Archer and Krishnamurthy (1993a, 1993b) systematically reviewed the Minnesota Multiphasic Personality Inventory (MMPI)-Rorschach research and found that there is minimal if any association between the two tests. In response to this review, data are reported demonstrating a positive association between various Rorschach variables and objective criteria. Data gleaned from published and yet unpublished sources suggest that the Rorschach is related to various self-report, other report, and behavioral criteria, with children and adults, for psychosis and other constructs. Data are also presented suggesting that part of the disagreement between the two tests may be due to different types of response bias and positive self-presentations on the two tests. Research findings pertinent to differential validity as a function of the number of Rorschach responses protocol are also explored. I conclude with research recommendations regarding test specific understandings of response bias and Rorschach protocol complexity as a moderator variable. PMID- 16367677 TI - Construct validation of scales derived from the Rorschach method: a review of issues and introduction to the Rorschach rating scale. AB - I suggest the main goal of Rorschach validation should be a refined understanding of what each score means. Toward this end, I review general issues in construct validity, hurdles unique to the Rorschach, and general limitations with validation criteria. I then recommend two approaches for improving criteria so they can begin approximating the gold standards that are necessary for a refined understanding of what scores actually measure. The first is a method for improving expert clinical judgment, and the second is a method for aggregating data across diverse judges. Finally, the Rorschach Rating Scale (RRS) is presented as a criterion tool to be used with either of these approaches to validation. The RRS is a fairly comprehensive summary of the constructs thought to be measured by various Rorschach scoring systems. The utility of the RRS for research and training are discussed, as are other practical, theoretical, and psychometric issues in its application. PMID- 16367678 TI - Personal problem-solving scoring of TAT responses: known-groups validation. AB - Two studies employed a known-groups validation strategy to evaluate a Thematic Apperception Test (TAT) scoring system purported to measure personal problem solving skills, the Personal Problem-Solving System (PPSS). In Study 1 clinicians rated the records of 46 mental health outpatients for the presence of personal problem-solving skills deficits. Verbatim TAT transcripts from these patients were blindly scored using the PPSS. Participants predicted to demonstrate problem solving deficits obtained lower PPSS scores. In Study 2 a psychiatric sample (n = 47) and a community-based comparison group (n = 47) completed a life history questionnaire, a checklist of problems currently experienced, a measure of psychiatric symptoms, and responded to 3 TAT cards. TAT responses were again blindly scored using the PPSS. In contrast to the comparison group, psychiatric patients checked a greater number of current problems, endorsed more psychiatric symptoms, and obtained lower scores on the PPSS. A discriminant function analysis using PPSS scores correctly classified 72% of these participants. PPSS scores predicted group membership even after controlling for differences in age, education, the number of problems experienced, and psychiatric symptoms. This combination of variables correctly classified 92% of the participants. Results of both studies are interpreted as supporting the discriminant validity of the PPSS. PMID- 16367681 TI - Testing the bipolarity of the Jungian functions. AB - In this article, I offer an alternative explanation for recent studies that have questioned the assumed bipolarity of the Jungian functions measured by the Myers Briggs Type Indicator (MBTI) due to the failure to observe negative correlations between the hypothesized opposite functions when a Likert format is used in place of the MBTI's forced-choice format (Cowan, 1989; Girelli & Stake, 1993; Loomis & Singer, 1980). It was suggested that the acquiescence response set might be partially responsible. A new instrument was constructed using agree-disagree items. Negative correlations between the hypothesized opposite functions were not observed. To deal with the acquiescence response set, participants were ranked on the number of statements endorsed, and the scores for the middle third of participants were analyzed again For these participants, correlations of -.67 and -.69 between each pair of theoretically opposite functions were found. I conclude thai this largely refutes criticisms of the bipolarity assumption, but there is still reason to develop a measure not relying on the assumption. PMID- 16367682 TI - The assessment of trust orientation. AB - The development of the Trust Inventory, a 40-item measure of trust in generalized others (Generalized Trust) and romantic partners (Partner Trust) is described. A third conceptualization of trust in friends and family members (Network Trust) is also discussed College students (N = 1,229) participated in five stages of test construction and validation. Results indicated that the Trust Inventory scales are reliable, both internally and temporally, and that the Partner Trust and Generalized Trust Scales demonstrate both concurrent and construct validity. The resulting inventory is unique in its capacity to assess these types of trust simultaneously. Evidence supporting the discriminant validity of the Network Trust was mixed, whereas factor analytic treatments of Trust Inventory items supported the distinctiveness of Network Trust as compared to Partner and Generalized Trust, the Network Trust Scale correlated to roughly the same degree as the other two scales with several variables of differential theoretical relevance. Thus, little evidence supporting the incremental validity of Network Trust was observed Discussion focuses on the evidence suggesting the validity of interpretations of Generalized and Partner Trust and considers the issue of additional research regarding Network Trust. PMID- 16367683 TI - Searching for conscientiousness on the MMPI-2. AB - It has been argued that the revised Minnesota Multiphasic Personality Inventory (MMPI-2) fails to measure the Big Five dimension of conscientiousness (C) an assertion supported by the research finding that none of the factors underlying the MMPI correlate substantially with the C factor of the NEO Personality Inventory (NEO-PI)-a popular operationalization of the Big Five model. In this study we reconsidered the relationship between the MMPI-2 item pool and the C factor of the NEO-PI-R (Form S). Collections of MMPI-2 C markers did not correlate much higher than .5 with the NEO-PI-R factor scale in cross-validation hold-out samples. Most pervasive, however, was the finding that MMPI-2 item markers of C were also strongly (in many cases, more highly) related to the NEO PI-R factor of neuroticism (N). Efforts to purify collections of MMPI-2 C markers, first by careful item selection and then by suppression of N variance, were met with limited success. Intercorrelations among NEO-PI-R scales suggest that the difficulty in finding markers that discriminate between N and C extends beyond any shortcomings of the MMPI-2 item pool. PMID- 16367685 TI - Editor's note: series Coda. PMID- 16367684 TI - Is the Rorschach welcome in the courtroom? AB - A survey of 7,934. recent federal and state court cases in which psychologists presented Rorschach testimony revealed only 6 in which the appropriateness of the Rorschach was challenged and just one in which the testimony was not admitted into evidence. There is accordingly good reason to believe that, despite occasionally voiced concerns to the contrary, the Rorschach is welcome in the courtroom. PMID- 16367686 TI - Return to the source: Rorschach's psychodiagnostics. PMID- 16367690 TI - Culturally competent assessment practice in the United States. AB - Assessment practice with multicultural populations in the United States requires cultural competence. Culturally competent assessment includes culture-specific styles of service delivery, use of the client's first language, and an evaluation of the client as a cultural being prior to test administration using cultural orientation categories. This information is essential for understanding health/illness beliefs, to indicate the possible presence of culture-specific symptoms, and to select standard or culture-specific tests. Assessment ethics recognize the desirability of alteration or modification of standard tests by translation and development of new norms. Cross-cultural construct validity, response sets, and cultural variables are also research priorities. Culture specific tests should be constructed and validated for some clients as determined by cultural orientation status. Feedback of test findings to clients and significant others should be consistent with cultural expectations. Professional and societal benefits of cultural competence in assessment are suggested. PMID- 16367691 TI - A new measure for an important construct: the attachment and object relations inventory. AB - This article describes the development of the Attachment and Object Relations Inventory (AORI). The AORI, a dimensional measure based on factor structure, was developed in response to the psychometric problems typical to attachment style measures that are categorical in nature. Test-retest reliability and internal consistency were high. A factor analysis of the 75 AORI items confirmed the existence of at least 6 factors and explained approximately 50% of the item variance. Convergent and discriminant validity of the AORI was supported through appropriate relationships with measures of depression, anxiety, and personality, and other measures of attachment and object relations. PMID- 16367692 TI - Reliability of an idiographic Q-sort measure of defense mechanisms. AB - Despite the important insights the concept of defense mechanisms may offer to our understanding of human behavior, no standardized definitions of defense mechanisms have been universally accepted. Inconsistencies in the definition and conceptualization of defense mechanisms has limited the practical utility of research involving these constructs. In addition, lack of interrater reliability, use of anecdotal evidence, and reliance on self-reports has retarded their investigation. Conducting methodologically rigorous investigations with a psychometrically sound instrument is the first step in addressing some of the issues concerning defense mechanisms and their theoretical postulates. This study was conducted to determine the reliability associated with the Defense-Q, an observer-based Q-sort measure of defense mechanisms. Thirty participants who had undergone an interpersonally stressful interview (the Type A Structured Interview; Rosenman, 1978) were rated by 11 trained coders, both for their use of the 25 defense mechanisms and for their ego strength. Reliability was assessed using Cronbach's alpha. Individual defense mechanisms demonstrated reliability ranging from .28 (undoing) to .92 (humor), with an average reliability of .73. Coder reliability ranged from .63 to .76, with an average of .69. These results indicate that defense mechanisms can be reliably assessed by the Defense-Q. Reliability of the Defense-Q is compared to existing observational measures of defense mechanisms. PMID- 16367693 TI - Evaluating the similarity of MMPI-2 and MMPI profiles: reply to Dahlstrom and Humphrey. AB - We review issues that have arisen in exchanges with Dahlstrom and Humphrey (Dahlstrom & Humphrey, 1996; Humphrey & Dahlstrom, 1995) about assessing the comparability of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) and the MMPI. We point out the limitations of Q correlations (without contending that D(2) is "the only legitimate function of profile comparability," as Dahlstrom and Humphrey, 1996, p. 350, claim we do), and explain why Dahlstrom and Humphrey's (1996) new Q-correlational results, correctly interpreted, are consistent with our own previous observations and conclusions. We stress again the importance of both overall profile elevation and profile "definition" in making code-type assignments. Nonrestrictive code types ignore these profile characteristics, and their use needlessly lowers MMPI-2/MMPI code-type congruences and raises the incidence of profile misinterpretations. Our recommendation of well-defined MMPI 2 code types stands. PMID- 16367694 TI - Relationships among affect in play, interpersonal themes in fantasy, and children's interpersonal behavior. AB - Relationships among children's affective expression in fantasy play, interpersonal themes in projective stories, and concurrent interpersonal behaviors were investigated in 49 children. Participants completed a play task, a projective storytelling task, and a brief IQ measure. Peer and teacher ratings provided measures of interpersonal functioning. Results found that access to and comfort with affect is related to the ability to think in interpersonal ways. Neither affective expression in play nor interpersonal themes in projective stories were related to actual interpersonal behavior. PMID- 16367695 TI - John Exner and the book that started it all: a review of The Rorschach Systems. AB - This article is a review and a reassessment of Exner's (1969) The Rorschach Systems. The book reviews the history of the development of the Rorschach, primarily in the United States, including a detailed analysis of the progressive development of systems devised by Klopfer, Beck, Piotrowski, Hertz, Rapaport, and Schafer. The final versions of each system are compared with each other, and with Rorschach's own system. The development of the book is linked to Exner' s personal relationship with Klopfer, Beck, and Hertz. The book is described as the basic forerunner of Exner's Comprehensive System. PMID- 16367699 TI - Introduction to the special series--interpersonal theory and the interpersonal circumplex: Timothy Leary's legacy. PMID- 16367700 TI - An informal history of the interpersonal circumplex tradition. AB - Circular representations of human nature can be traced from ancient astrology to Leonardo Da Vinci to the modern interpersonal circumplex (IPC) tradition. which began with Harry Stack Sullivan. The Kaiser Foundation Psychology Research Group operationalized key concepts of Sullivan's and Timothy Leary (1957) provided extensions of Sullivanian theory that formed the conceptual basis for contemporary circumplex models The current generation of circumplex investigators is exceptionally well represented by the contributors to this special series. The IPC can serve as an integrative cognitive framework for making sense of diverse realms that are well outside its original focus of convenience. The visual and psychological appeal of this powerful model is briefly illustrated. PMID- 16367701 TI - The interpersonal circle as a heuristic model for interpersonal research. AB - In this article, I review major advances in the development of the interpersonal circle and its measurement, as delineated by Timothy Leafy (1957) and his colleagues. In my early work (Lorr & McNair, 1963, 1965), an interpersonal Behavior Inventory consisting of manifest behavioral statements was constructed. The 14 categories were found to fit a circular order in several samples. Studies using LaForge and Suczek's (1955) Interpersonal Check List, and Wiggins's (1979) Interpersonal Adjective Scales further established the value of Leary's conceptions and extended them. Several major theorists, researchers, and clinicians including Benjamin, Carson, Horowitz, Kiesler, and Wiggins have been inspired by Leary and, in many cases, have extended his ideas in ways not originally envisioned. Recent research (e.g., Hofstee, de Raad, & Goldberg, 1992; Millon, 1987) demonstrated the pertinence of the interpersonal conception of personality to the 5-factor model and personality disorders. PMID- 16367702 TI - Interpersonal circumplex. AB - The social and intellectual climate of the late 1940s and early 1950s in America helped nourish humanistic, person-centered views of human behavior. During that time, psychologists such as Gordon Allport, Abraham Maslow, David McClelland, Harry Murray, and Carl Rogers emphasized the positive growth potential in human character. The psychiatrist Harry Stack Sullivan proposed that personality can best be understood within the context of interpersonal transactions, and he provided a practical, street-smart understanding of psychiatric symptoms that was quite an advance over the traditional medical and psychoanalytic viewpoints. These ideas, along with the concept of dimensionalizing traits rather than categorizing them, inspired my colleagues and I to conduct our cooperative work on the interpersonal circumplex, which culminated in the publication of my monograph. Interpersonal Diagnosis of Personality (Leary, 1957). PMID- 16367703 TI - Comparability of MMPI and MMPI-2 profile patterns: Ben-Porath and Tellegen's inappropriate invocation of Mahalanobis's D2 function. AB - Ben-Porath and Tellegen (1995) claimed that the data in the article by Humphrey and Dahlstrom (1995) were improperly analyzed by means of Q correlations between raw scores earned by the individuals in the forensic sample to establish pattern comparability between the original Minnesota Multiphasic Personality Inventory (MMPI; Hathaway & McKinley, 1943) and the Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989), which were then contrasted with Q correlations between the corresponding T-score patterns. Ben-Porath and Tellegen (1995) contended that the Q correlation is affected by random factors and that a generalized distance function, D2, is the only legitimate index of profile comparability. Data are presented here to show that the Q correlation serves as a reliable index of pattern comparability, relatively unaffected by differences in profile elevation. The Mahalanobis (1936) D2 index is too heavily weighted with differences in profile elevation serve as the proper index of equivalence in profile patterning. The findings in the Humphrey and Dahlstrom (1995) article were based on appropriate data-analytic procedures because the primary concern is their investigation was the extent to which the patterns of T-score profiles from the original MMPI and the MMPI-2 are comparable when the raw-score patterns are virtually identical. PMID- 16367707 TI - MMPI-2 clinical correlates for ten common codes. AB - A number of researchers have called for the establishment of a Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) correlate literature that is based directly on investigations of the revised MMPI. The purpose of this study was to examine clinical correlates found for 9 commonly occurring 2-point codes, each of which contained a minimum of 20 patients, and for 82 profiles that were within-normal limit (WNL) in a sample of 289 female and 308 male psychiatric inpatients. Major findings indicated that descriptors identified for MMPI-2 based codes were generally similar to the established literature for corresponding codes derived from the original MMPI (Hathaway & McKinley, 1967). In addition, correlates identified for the WNL profile suggest that these patients are less likely to exhibit more chronic symptomatology and to have psychological histories marked by less severe psychological disturbance than other inpatients. Overall, the results of this study represent an initial step in the creation of an MMPI-2 correlate literature. PMID- 16367708 TI - Detecting fake-bad MMPI-A profiles. AB - This study examined the extent to which the validity scales of the Minnesota Multiphasic Personality Inventory-Adolescent (MMPI-A; Butcher et al., 1992) could identify adolescents who were faking-bad. This study also examined the extent to which the validity scales could differentiate between nonclinical adolescents instructed to fake-bad and both clinical and nonclinical adolescents who received standard instructions. Participants were 58 male and 80 female high school students and 58 male and 80 female adolescents from clinical settings. Results indicated that the mean profiles resembled those previously reported for adults on the MMPI (Graham, 1987) and MMPI-2 (Graham, Watts, & Timbrook, 1991) and for adolescents on the original MMPI (Archer, Gordon, & Kirchner, 1987). Accurate identification of students who were faking-bad was achieved. It was possible to differentiate between the clinical adolescents and nonclinical adolescents who were faking, and it was possible to differentiate between nonclinical adolescents who were faking and nonclinical adolescents with standard instructions; however, different cutoff scores were needed for these two discriminations. Optimal cutoff scores were presented. PMID- 16367709 TI - Validation of the Clance Imposter Phenomenon Scale. AB - The Clance Impostor Phenomenon Scale (CIPS; Clance, 1985) was compared to the newly developed Perceived Fraudulence Scale (Kolligian & Sternberg, 1991). The two scales were found to have high internal consistency and to correlate in a similar manner with other measures. Further, discriminant validity evidence for the Impostor Phenomenon (IP) was provided by comparing the CIPS to measures of depression, self-esteem, social anxiety, and self-monitoring. The IP was related to, but substantially discriminable from, these constructs. Finally, construct validity evidence for the CIPS was provided through principal components analysis that yielded three stable factors: Fake, Discount, and Luck. PMID- 16367710 TI - Personological evaluation of Clance's Imposter Phenomenon Scale in a Korean sample. AB - The purpose of this study was both to determine if the Impostor Phenomenon (IP) can be reliably and validly assessed in a Korean context and if so, evaluate the construct within the context of Jungian typology and the 5-factor model of personality. A sample of 654 Korean men and women were selected from 4 major Korean cities and administered the Clance Impostor Phenomenon Scale (CIPS; Clance & Imes, 1978) along with the Myers-Briggs Type Indicator (MBTI, Form G; Myers & McCaulley, 1985) and NEO Personality Inventory-Revised (NEO-PI-R; Costa & McCrae, 1992). Results indicated that the CIPS was very reliable, and the pattern of correlates suggested impostors to be introverted types on the MBTI. Results with the NEO-PI-R showed impostors to be very high on neuroticism and low on conscientiousness. This pattern of correlates is similar to other performance inhibiting constructs such as fear of success and fear of failure. It was argued that IP be construed more as a motivational style than as a distinct clinical syndrome. The IP seems to be less pervasive in Korea than America and these cross cultural implications were discussed. PMID- 16367711 TI - The attention to body shape scale: a new measure of body focus. AB - An abnormal degree of attention to one's body shape has been implicated in the development and maintenance of eating pathology. This article reports on three preliminary reliability and validity studies of the Attention to Body Shape Scale (ABS). Although brief, the ABS was found to be internally consistent and relatively stable over 2 weeks for both women and men. As expected, women were more body-focused on the ABS than were men. Preliminary validation studies on college women had encouraging results. Lending evidence of convergent validity, the ABS correlated with measures of eating pathology and dietary restraint. Moreover, although the ABS correlated with a measure of general-appearance orientation, regression analyses showed that the former was more closely linked than the latter to eating pathology. As evidence of discriminant validity, the ABS failed to correlate with dysphoria, body weight, or the tendency to eat in response to emotional or environmental pressures. As predicted by theory, high scores on the ABS were associated with body-image distortion. However, contrary to predictions, the ABS failed to moderate the impact of induced anxiety on body image on two of three measures. The implications of these findings and suggestions for future work are discussed. PMID- 16367712 TI - Maximizing internal and external validity in MMPI malingering research: a study of a military population. AB - The authors investigated the effectiveness of various commonly used Minnesota Multiphasic Personality Inventory (MMPI; Hathaway & McKinley, 1943) indices of exaggeration and malingering in detecting suspected malingering in a military sample of 121 enlisted men. To maximize external validity, only men undergoing psychological evaluation were used as participants. Forty-one participants were identified as suspected malingerers through multiple criteria and were contrasted with schizophrenic-spectrum and clinic outpatient groups. To improve internal validity, the 41 suspected malingering participants were asked to retake the test without exaggerating. Results revealed that there were many false positives and fewer, but nonetheless many, false negatives with standard malingering indices. It appeared that the Gough Dissimulation scale (Gough, 1947) might hold the most promise as a measure of malingering, but other scales are also useful. Individual comparisons between different samples and implications for MMPI-2 (Butcher et al., 1989) are presented. PMID- 16367713 TI - Comparing three new MMPI-2 randomness indices in a novel procedure for random profile derivation. AB - This study compared three new validity indices; /F-Fb/, VRIN+/F-Fb/, and F+Fb+/F Fb/; to the F, back-F, and VRIN according to their effectiveness in detecting degrees of profile randomness. Participants completed the Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, and Kaemmer, 1989) according to standardized instructions. All profiles starting at Items 142, 284, or 426 were substituted with computer-generated random responses to produce 75%, 50%, and 25% random profiles, respectively. Twenty-five profiles were derived using 100% computer-generated items. Finally, one unaltered set of profiles was designated as 0% random. This generated five groups of MMPI-2 profiles, differing according to the degree of profile randomness (i.e., 0%, 25%, 50%, 75%, and 100%). Results showed that the F and /F-Fb/ indices were unable to distinguish authentic from all degrees of randomness, whereas the F+Fb+F-Fb scale could reliably distinguish all levels of randomness. The actuarial analysis found back-F and F+Fb+/F-Fb/ misclassified the least number of profiles, suggesting the latter index should be included in the evaluation of MMPI-2 profile randomness. PMID- 16367716 TI - Ideographic aspects of injury memories: applying contextual theory to the Comprehensive Early Memories Scoring System--revised. PMID- 16367714 TI - Psychometric characteristics of the MMPI-2 Cook-Medley Hostility scale. AB - The Minnesota Multiphasic Personality Inventory (MMPI; Hathaway & McKinley, 1943) Cook-Medley Hostility scale (Ho) has been studied a great deal because of its relation to coronary disease and mortality. However, little research has been conducted with the Ho scale on the revised MMPI (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989). This study examined the psychometric characteristics of the MMPI-2 Ho scale. Only nine of the original 50 Ho items were slightly changed in the revision. Ho scores were highly correlated with MMPI 2 scales CYN, K, TPA, and ASP, supporting the interpretation of Ho as a measure of cynicism. High correlations with other MMPI-2 scales also suggested that Ho is related to general psychopathology and negative affectivity. Male Ho scorers were rated by their spouses as hotheaded, bossy, demanding, and argumentative. For women, Ho scale scores were less strongly associated with ratings of overt hostility. Principal components analysis of Ho revealed four underlying dimensions: Cynicism, Hypersensitivity, Aggressive Responding, and Social Avoidance. PMID- 16367717 TI - Origin and history of the Thematic Apperception Test images. AB - The origin and history of each of the images appearing on the 1943/1971 edition of the Thematic Apperception Test is presented. This information corrects and supplements that provided by Murray (1943/1971) in the test manual. Authorship of the test, including the contributions of Christiana Morgan, is discussed. The material presented expands the historical record and may serve as an aid in test interpretation. PMID- 16367718 TI - Differential relationships of objective and projective dependency scores to self reports of interpersonal life events in college student subjects. AB - Eighty-three undergraduate subjects (58 women and 25 men) participated in a prospective study in which they (a) completed widely used objective and projective measures of dependency, and then (b) provided monthly reports of the frequency and impact of various types of life events during a 1-semester (3 month) period. As expected, subjects' projective dependency scores predicted their frequency estimates and impact ratings of interpersonal life events but were unrelated to frequency estimates and impact ratings of other types of life events (e.g., achievement-related, legal). Objective dependency scores were unrelated to all life event frequency estimates and impact ratings. Findings are discussed in the context of recent theoretical frameworks that distinguish implicit dependency needs (which are assessed via projective measures) from self attributed dependency needs (which are assessed via self-report tests). The importance of the type of dependency measure used in studies of the dependency life events relationship is emphasized. PMID- 16367719 TI - Validity of the MMPI subtle items. AB - This article presents a review and critique of research on the Minnesota Multiphasic Personality Inventory (MMPI) subtle items in regard to their validity as measures of their scales' characteristics. The main categories of focus are their convergent and discriminative validity, the paradoxical effect, and the issue of whether removing these items would improve MMPI validity. Important problems in the methodologies are noted and discussed in some detail. These methodological concerns and the conflicting results across studies obscure firm conclusions on the validity of the subtle items, although strong support is lacking. However, more evidence is needed before deciding to remove these items from the MMPI; therefore, this article provides recommendations for future research. PMID- 16367720 TI - Psychological implications of the Psychopathy Q-sort. AB - The construct validity of the Psychopathy Q-sort (PQS; Reise & Oliver, 1994) was investigated by correlating it with an array of observer-based and self-report personality measures in a sample of 350 men and women assessed at the Institute of Personality Assessment and Research (IPAR). High discriminant validity of the PQS was indicated by a pattern of significant correlations with self-report measures of the Cluster B personality disorder scales of the Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev.; American Psychiatric Association, 1980) and the absence of correlations with scales belonging to Clusters A and C of the manual. The PQS also correlated positively with the California Psychological Inventory measures of social poise and assurance and correlated negatively with measures of normative control of impulse. Gender differences in psychopathy were explored using Adjective Check List (Gough & Heilbrun, 1983) ratings provided by IPAR staff observers and by 76 pairs of spouses. PMID- 16367721 TI - Comparability and validity of computerized adaptive testing with the MMPI-2. AB - The comparability and validity of a computerized adaptive (CA) Minnesota Multiphasic Personality Inventory-2 (MMPI-2) were assessed in a sample of 571 undergraduate college students. The CA MMPI-2 administered adaptively Scales L, E the 10 clinical scales, and the 15 content scales, utilizing the countdown method (Butcher, Keller, & Bacon, 1985). All subjects completed the MMPI-2 twice, with three experimental conditions: booklet test-retest, booklet-CA, and conventional computerized (CC)-CA. Profiles across administration modalities show a high degree of similarity, providing evidence for the comparability of the three forms. Correlations between MMPI-2 scales and other psychometric measures (Beck Depression Inventory; Symptom Checklist-Revised; State-Trait Anxiety and Anger Scales; and the Anger Expression Scale) support the validity of the CA MMPI-2. Substantial item savings may be realized with the implementation of the countdown procedure. PMID- 16367722 TI - The development and validation of the Body-Image Ideals Questionnaire. AB - The Body-Image Ideals Questionnaire (BIQ) was developed as a unique attitudinal body-image assessment that considers one's perceived discrepancy from and degree of investment in personal ideals on multiple physical attributes. Reliability and validity of the 20-item instrument were examined for a sample of 284 college women. The results indicated that the BIQ consists of two relatively distinct and internally consistent Discrepancy and Importance subscales, as well as their multiplicative composite. The subscales' respective convergent validities vis-a vis extant body-image measures and specific facets of personality (i.e., public self-consciousness and perfectionism) and psychosocial adjustment (i.e., social anxiety, depression, and eating disturbance) were confirmed. Evidence also supported the incremental validity of multiple self-ideal discrepancies. Effects due to socially desirable responding were inconsequential. Directions for needed basic and clinical research were identified. PMID- 16367723 TI - Clarifying body-image disturbance: analysis of a multidimensional model using structural modeling. AB - A multidimensional model of body-image disturbance was tested. The model incorporated the concepts of body-size distortion, preference for thinness, body dissatisfaction, and fear of fatness as predictors of restrictive eating. The LISREL 7 program was used to perform a structural modeling analysis of the theoretical model. A total of 175 women participated in the study (54 eating disordered patients and 121 undergraduate students). The results supported the hypothesized four-dimensional model relative to alternative one-, two-, and three dimensional models. Body dissatisfaction appeared to be directly affected by both body-size distortion and preference for thinness. Fear of fatness was found to be the best predictor of restrictive eating. The results appeared consistent across the clinical and nonclinical samples. These data may help resolve many of the current controversies in the body-image literature. The results also suggested the need to develop more sound assessment instruments for fear of fatness. PMID- 16367726 TI - The Rorschach: projective technique or psychometric test? AB - Various approaches to the Rorschach Technique are described in terms of the idiographic-nomothetic axis and the perceptual-content axis. It is suggested that it is most productive to view the Rorschach as a projective tool, with perceptual scoring a secondary factor. Current efforts at objectification of the Rorschach are not seen as useful as efforts to enhance its projective qualities. Some possible ways are discussed in which the projective value of the instrument can be maximized. PMID- 16367728 TI - Integrative Rorschach interpretation. PMID- 16367727 TI - Putting your eggs in the content analysis basket: a response to Aronow, Reznikoff, and Moreland. AB - A response is made to the essay by Aronow, Reznikoff, and Moreland (this issue) on the superiority of the content analysis approach to the Rorschach. Their argument is seen as giving insufficient emphasis to the importance of information obtained from empirically based, quantified methods of Rorschach interpretation. Content analysis without score-based interpretation is not sufficiently reliable for clinical use. Also, the argument of Aronow and colleagues that the Rorschach is predominantly a projective technique is refuted. Rorschach interpretation that deemphasizes empirically based scores is as insufficient as interpretation that excludes content analysis. PMID- 16367730 TI - A survey of graduate students' experiences, interests, and attitudes about learning the Rorschach. AB - A questionnaire about the experiences, interests, and attitudes of graduate students learning the Rorschach was sent to 235 student affiliates of the Society for Personality Assessment. The questionnaire asked for information concerning their Rorschach training in (a) class instruction, (b) adjunctive instruction (i.e., extraclass supervision and experiences at practicum sites), (c) research, and (d) personal experiences and views concerning this training. Seventy-one percent (166) of the questionnaires were returned. Results indicate that almost half the respondents gave from zero to four Rorschachs in their first course, that only half the students took a second course on the Rorschach, and that the Exner Comprehensive System was learned by a majority of the students. Students felt somewhat unsure of their skills concerning the formulation of Rorschach findings into concepts that could be communicated in a written report. Students reported that they lacked instruction concerning the research findings and application of the Rorschach. Students also did not feel grounded well enough in theory. Specific recommendations based on the information gathered from this survey are offered to improve teaching practices in the Rorschach as well as in personality-assessment training. PMID- 16367731 TI - Pigeons, people, and pigeon-holes. AB - Contemporary theorizing about the nature of personality has neglected typological formulations. Instead, reliance has been placed on multidimensional geometric models that fail to capture the crucial configural nature of personality structure and functioning. As a result, there has been little progress in the development of a comprehensive taxonomy of human personalities or in the establishment of a personological systematics. There is evidence that many psychodiagnostic formulations rely on implicit sets of categories or taxa of individuals together with the various forms of psychopathology that are associated with these types. Some reasons for this neglect are pointed out, related to a general aversion to "pigeon-holing" people and the risk of applying stereotypes rather than theorotypes. However, the recent development of powerful taxometric procedures for locating personological taxa is noted and the potential benefits for the science and art of personality assessment to be gained from a comprehensive personological taxonomy are discussed. PMID- 16367732 TI - Domains and facets: hierarchical personality assessment using the revised NEO personality inventory. AB - Personality traits are organized hierarchically, with narrow, specific traits combining to define broad, global factors. The Revised NEO Personality Inventory (NEO-PI-R; Costa & McCrae, 1992c) assesses personality at both levels, with six specific facet scales in each of five broad domains. This article describes conceptual issues in specifying facets of a domain and reports evidence on the validity of NEO-PI-R facet scales. Facet analysis-the interpretation of a scale in terms of the specific facets with which it correlates-is illustrated using alternative measures of the five-factor model and occupational scales. Finally, the hierarchical interpretation of personality profiles is discussed. Interpretation on the domain level yields a rapid understanding of the individual interpretation of specific facet scales gives a more detailed assessment. PMID- 16367733 TI - A new method for detecting brain disorder by measuring perseveration in personality inventory responses. AB - A 130-item personality inventory (the Psychological Screening Inventory, Lanyon, 1970) was used to obtain a novel measure of perseveration coupled with affirmative set. Acquiescent Perseveration (AP), a quantitative behavioral metric, is the sum of the lengths of True (T) response runs 4 and longer, doubling T runs 9 and longer. With male subjects, a one-way ANOVA comparison of heterogeneous organics with five non-organic control groups (normals, neurotics, sociopaths, personality disorders, and schizophrenics) showed mean AP differences greater than .001 versus every control group. The measure, requiring no scoring key, was successfully cross-validated with six similar female groups. AP means for non-organics showed no increase across 5 decades of age. In small supplementary studies AP was related inversely to Shipley Abstraction (Shipley, 1940, 1946) scores and directly to the Halstead-Reitan Impairment Index (Reitan & Davison, 1974). AP, uncorrected for age, IQ or education, matches the best single cognitive neuropsychological tests in distinguishing organics from psychiatric patients. Inventory response sequences contain previously unrecognized latent data sensitive to neuropathology. PMID- 16367734 TI - A five-year follow-up on the temporal stability of the Ego Impairment Index. AB - This study investigated the temporal stability of the Ego Impairment Index (EII) over a 5-year period. The EII is based on the Comprehensive System (Exner, 1993) and has shown promise as a measure of internal disturbance and psychopathology (Perry & Braff, 1994; Perry & Viglione, 1991; Perry, Viglione, & Braff, 1992). This study successfully recalled 17 subjects from an original sample of 46 who initially suffered from Major Depression, Melancholic-Type 5 years earlier and were treated with antidepressant medication. These subjects did not differ significantly from those Subjects who were not successfully recalled. The Rorschach and a variety of other measures were administered to the subjects. The results revealed impressive temporal consistency over 5 years: a rank-order, test retest correlation of .68. The EII was also correlated with some of the measures of overall adaptation. These results are consistent with the notion that the EII is a stable trait-measure of psychopathology. Limitations because of the small subgroup of subjects, although not unusual in longitudinal research, are noted. PMID- 16367735 TI - Personal problem-solving scoring of the TAT: sensitivity to training. AB - The Thematic Apperception Test (TAT; Murray, 1943) is one of the most frequently taught and administered projective instruments in the United States. Nevertheless, formal scoring systems are not often used when evaluating story responses; thus, the reliability and validity of response interpretations remain controversial. The sensitivity-to-training of a procedure that uses the TAT to assess personal problem-solving skills, namely the Personal Problem-Solving System (PPSS; Ronan, 1990), was evaluated. In Experiment 1, undergraduate students received either detailed (n = 30) or minimal (n = 29) training in the problem-solving skill of generating alternative solutions to personal problems and subsequently wrote responses to three TAT cards. In Experiment 2, subjects received either detailed (n = 22) or minimal (n = 22) training in a utility model of decision making and thereafter responded to the same three TAT cards. Subjects who received detailed training, when compared to their minimally trained counterparts, reported an increase in their knowledge of the relevant problem solving skill, and this increased knowledge influenced their responses to the TAT cards as measured by the PPSS scores. Implications for the construct validity of the PPSS are discussed. PMID- 16367736 TI - Comparing five alternative factor-models of the Student Jenkins Activity Survey: separating the wheat from the chaff. AB - The Student Jenkins Activity Survey (SJAS; Glass, 1977) is the most popular instrument for measuring Type A behavior in college undergraduates, and a variety of different unidimensional and multidimensional models for scoring the SJAS have been employed in the past. This study compared five alternative factor-models developed for the short (21-item) form of the SJAS in terms of their overall goodness of fit, factorial invariance, and factor reliabilities, using the data of two independent samples of undergraduates. Indices of relative fit for the five models ranged from .64 to .94 (median = .85), but only one unidimensional model and one multidimensional model achieved a minimum satisfactory goodness of fit (.90 or greater) for both samples. Reconceptualizing these models within a broader nomological net sheds light on their meaning and highlights existing gaps in the measurement of multifaceted Type A behavior. PMID- 16367738 TI - Differential convergence of self-report and informant data for multidimensional personality questionnaire traits: implications for the construct of negative emotionality. AB - Construct validation work on Tellegen's (1982) Multidimensional Personality Questionnaire (MPQ) resulted in further inferences about Negative Emotionality. Two hundred thirty-two students were rated by three knowledgeable informants, yielding a total of 928 participants. The median monotrait correlation of MPQ primary scores with summed observer ratings was .48, and all were significant, p < .01. These data show higher self-report to informant rating convergences in the Positive Emotionality (Extroversion) domain than in the Negative Emotionality (Neuroticism) domain. Furthermore, in the Negative Emotionality domain, peers, mothers, and fathers were not equivalent as classes of raters. Stress Reaction ratings showed uniformly lower levels of convergence with self-report (relative to Positive Emotionality traits) across all rater classes. For Alienation, peer and maternal ratings were comparable, but paternal ratings correlated significantly less with self-report scores than did maternal ratings. And finally, with Aggression, peer ratings correlated significantly higher with self report than either maternal or paternal ratings. These findings, taken in the context of the literature, have implications for a hierarchical model of Negative Emotionality, support inferences about the display of cues of Negative Emotionality, and offer new cautions for rating-based assessment. PMID- 16367739 TI - Intracellular localization of p40, a protein identified in a preparation of lysosomal membranes. AB - Unlike lysosomal soluble proteins, few lysosomal membrane proteins have been identified. Rat liver lysosomes were purified by centrifugation on a Nycodenz density gradient. The most hydrophobic proteins were extracted from the lysosome membrane preparation and were identified by MS. We focused our attention on a protein of approx. 40 kDa, p40, which contains seven to ten putative transmembrane domains and four lysosomal consensus sorting motifs in its sequence. Knowing that preparations of lysosomes obtained by centrifugation always contain contaminant membranes, we combined biochemical and morphological methods to analyse the subcellular localization of p40. The results of subcellular fractionation of mouse liver homogenates validate the lysosomal residence of p40. In particular, a density shift of lysosomes induced by Triton WR-1339 similarly affected the distributions of p40 and beta-galactosidase, a lysosomal marker protein. We confirmed by fluorescence microscopy on eukaryotic cells transfected with p40 or p40-GFP (green fluorescent protein) constructs that p40 is localized in lysosomes. A first molecular characterization of p40 in transfected Cos-7 cells revealed that it is an unglycosylated protein tightly associated with membranes. Taken together, our results strongly support the hypothesis that p40 is an authentic lysosomal membrane protein. PMID- 16367740 TI - Hepatitis C virus (HCV) core protein enhances the immunogenicity of a co delivered DNA vaccine encoding HCV structural antigens in mice. AB - In the present study, recombinant HCV (hepatitis C virus) core proteins enhanced the immune response elicited by a co-delivered DNA vaccine encoding HCV core and envelope proteins. A mixture of the plasmid pIDKE2 and Co.173, a protein comprising the first 173 amino acids of HCV core, in particular induces a strong humoral response, including antibodies that recognized peptides representing hypervariable region I from different viral isolates. Moreover, positive lymphoproliferative responses against the HCV structural proteins, encoded by the plasmid, were detected after two doses with this mixture. When the HCV core protein used in the mixture with pIDKE2 was Co.120, a protein comprising the first 120 amino acids of the viral antigen, a strong humoral response and a positive lymphoproliferative response were also detected. The effectiveness of this formulation was tested in vivo by measuring the protection against infection with a recombinant vaccinia virus expressing HCV core protein. A 2 log reduction in vaccinia-virus titre was observed in mice immunized with the mixture of pIDKE2 and Co.120. Humoral and cellular immune responses elicited for the mixture of pIDKE2 with either Co.173 or Co.120 was stronger and more diverse than those generated by individual components. In conclusion, our results indicate that formulations comprising both DNA constructs and protein subunit vaccine candidates are able to elicit strong humoral and cellular immunity against several antigens. Particularly, HCV core protein might be used as a feasible vehicle/adjuvant for DNA vaccines. PMID- 16367741 TI - Molecular and functional characterization of a novel gonadotropin-releasing hormone receptor isolated from the common octopus (Octopus vulgaris). AB - GnRH (gonadotropin-releasing hormone) plays a pivotal role in the regulation of reproduction in vertebrates through interaction with a specific receptor. Previously, we isolated a GnRH homologue, oct-GnRH, from the common octopus (Octopus vulgaris). In the present study, we have identified a GnRH receptor (oct GnRHR) specific for oct-GnRH from Octopus brain. Oct-GnRHR includes domains and motifs typical of vertebrate GnRH receptors. The intron-inserted positions are conserved between oct-GnRHR and the chordate GnRHR genes. The oct-GnRHR expressed in Xenopus (South African clawed frog) oocytes was responsive to oct-GnRH, but not to any other HPLC fractions of the Octopus brain extract. These results show that oct-GnRHR is an authentic receptor for oct-GnRH. Southern blotting of reverse-transcription PCR products revealed that the oct-GnRHR mRNA was widely distributed in the central and peripheral nervous systems and in several peripheral tissues. In situ hybridization showed that oct-GnRHR mRNA was expressed in some regions involved in autonomic functions, feeding, memory and movement. Oct-GnRH was shown to induce steroidogenesis of testosterone, progesterone and 17beta-oestradiol in Octopus ovary and testis, where oct-GnRHR was abundantly expressed. These results suggest that oct-GnRH, like its vertebrate counterparts, acts as a multifunctional neurotransmitter, neuromodulator and hormone-like factor, both in Octopus central nervous system and peripheral tissues, and that both structure and functions of the GnRH family are, at least partially, evolutionarily conserved between octopuses and chordates. PMID- 16367742 TI - Heterologous production in Wolinella succinogenes and characterization of the quinol:fumarate reductase enzymes from Helicobacter pylori and Campylobacter jejuni. AB - The epsilon-proteobacteria Helicobacter pylori and Campylobacter jejuni are both human pathogens. They colonize mucosal surfaces causing severe diseases. The membrane protein complex QFR (quinol:fumarate reductase) from H. pylori has previously been established as a potential drug target, and the same is likely for the QFR from C. jejuni. In the present paper, we describe the cloning of the QFR operons from the two pathogenic bacteria H. pylori and C. jejuni and their expression in Wolinella succinogenes, a non-pathogenic -proteobacterium. To our knowledge, this is the first documentation of heterologous membrane protein production in W. succinogenes. We demonstrate that the replacement of the homologous enzyme from W. succinogenes with the heterologous enzymes yields mutants where fumarate respiration is fully functional. We have isolated and characterized the heterologous QFR enzymes. The high quality of the enzyme preparation enabled us to determine unequivocally by analytical ultracentrifugation the homodimeric state of the three detergent-solubilized heterotrimeric QFR enzymes, to accurately determine the different oxidation reduction ('redox') midpoint potentials of the six prosthetic groups, the Michaelis constants for the quinol substrate, maximal enzymatic activities and the characterization of three different anti-helminths previously suggested to be inhibitors of the QFR enzymes from H. pylori and C. jejuni. This characterization allows, for the first time, a detailed comparison of the QFR enzymes from C. jejuni and H. pylori with that of W. succinogenes. PMID- 16367743 TI - Cleavage of focal adhesion proteins and PKCdelta during lovastatin-induced apoptosis in spontaneously immortalized rat brain neuroblasts. AB - We have previously shown that lovastatin induces apoptosis in spontaneously immortalized rat brain neuroblasts. Focal adhesion proteins and protein kinase Cdelta (PKCdelta) have been implicated in the regulation of apoptosis. We found that lovastatin exposure induced focal adhesion kinase, Crk-associated substrate (p130(Cas)), PKCdelta cleavage and caspase-3 activation in a concentration dependent manner. Lovastatin effects were fully prevented by mevalonate. The cleavage of p130(Cas) was almost completely inhibited by z-DEVD-fmk, a specific caspase-3 inhibitor, and z-VAD-fmk, a broad spectrum caspase inhibitor, indicating that cleavage is mediated by caspase-3. In contrast, the lovastatin induced cleavage of PKCdelta was only blocked by z-VAD-fmk suggesting that PKCdelta cleavage is caspase-dependent but caspase-3-independent. Additionally, z VAD-fmk partially prevented lovastatin-induced neuroblast apoptosis. The present data show that lovastatin may induce neuroblast apoptosis by both caspase dependent and independent pathways. These findings may suggest that the caspase dependent component leading to the neuroblast cell death is likely to involve the cleavage of focal adhesion proteins and PKCdelta, which may be partially responsible for some biochemical features of neuroblast apoptosis induced by lovastatin. PMID- 16367744 TI - Effect of mutations K97A and E128A on RNA binding and self assembly of papaya mosaic potexvirus coat protein. AB - Papaya mosaic potexvirus (PapMV) coat protein (CP) was expressed (CPdeltaN5) in Escherichia coli and showed to self assemble into nucleocapsid like particles (NLPs). Twenty per cent of the purified protein was found as NLPs of 50 nm in length and 80% was found as a multimer of 450 kDa (20 subunits) arranged in a disk. Two mutants in the RNA binding domain of the PapMV CP, K97A and E128A showed interesting properties. The proteins of both mutants could be easily purified and CD spectra of these proteins showed secondary and tertiary structures similar to the WT protein. The mutant K97A was unable to self assemble and bind RNA. On the contrary, the mutant E128A showed an improved affinity for RNA and self assembled more efficiently in NLPs. E128A NLPs were longer (150 nm) than the recombinant CPdeltaN5 and 100% percent of the protein was found as NLPs in bacteria. E128A NLPs were more resistant to digestion by trypsin than the CPdeltaN5 but were more sensitive to denaturation by heat. We discuss the possible role of K97 and E128 in the assembly of PapMV. PMID- 16367745 TI - The proline-rich protein palladin is a binding partner for profilin. AB - Palladin is an actin-associated protein that has been suggested to play critical roles in establishing cell morphology and maintaining cytoskeletal organization in a wide variety of cell types. Palladin has been shown previously to bind directly to three different actin-binding proteins vasodilator-stimulated phosphoprotein (VASP), alpha-actinin and ezrin, suggesting that it functions as an organizing unit that recruits actin-regulatory proteins to specific subcellular sites. Palladin contains sequences resembling a motif known to bind profilin. Here, we demonstrate that palladin is a binding partner for profilin, interacting with profilin via a poly proline-containing sequence in the amino terminal half of palladin. Double-label immunofluorescence staining shows that palladin and profilin partially colocalize in actin-rich structures in cultured astrocytes. Our results suggest that palladin may play an important role in recruiting profilin to sites of actin dynamics. PMID- 16367746 TI - Diversity and junction residues as hotspots of binding energy in an antibody neutralizing the dengue virus. AB - Dengue is a re-emerging viral disease, affecting approx. 100 million individuals annually. The monoclonal antibody mAb4E11 neutralizes the four serotypes of the dengue virus, but not other flaviviruses. Its epitope is included within the highly immunogenic domain 3 of the envelope glycoprotein E. To understand the favorable properties of recognition between mAb4E11 and the virus, we recreated the genetic events that led to mAb4E11 during an immune response and performed an alanine scanning mutagenesis of its third hypervariable loops (H-CDR3 and L CDR3). The affinities between 16 mutant Fab fragments and the viral antigen (serotype 1) were measured by a competition ELISA in solution and their kinetics of interaction by surface plasmon resonance. The diversity and junction residues of mAb4E11 (D segment; V(H)-D, D-J(H) and V(L)-J(L) junctions) constituted major hotspots of interaction energy. Two residues from the D segment (H-Trp96 and H Glu97) provided > 85% of the free energy of interaction and were highly accessible to the solvent in a three-dimensional model of mAb4E11. Changes of residues (L-Arg90 and L-Pro95) that statistically do not participate in the contacts between antibodies and antigens but determine the structure of L-CDR3, decreased the affinity between mAb4E11 and its antigen. Changes of L-Pro95 and other neutral residues strongly decreased the rate of association, possibly by perturbing the topology of the electrostatic field of the antibody. These data will help to improve the properties of mAb4E11 for therapeutic applications and map its epitope precisely. PMID- 16367747 TI - Characterization of a recombinantly expressed proteinase K-like enzyme from a psychrotrophic Serratia sp. AB - The gene encoding a peptidase that belongs to the proteinase K family of serine peptidases has been identified from a psychrotrophic Serratia sp., and cloned and expressed in Escherichia coli. The gene has 1890 base pairs and encodes a precursor protein of 629 amino acids with a theoretical molecular mass of 65.5 kDa. Sequence analysis suggests that the peptidase consists of a prepro region, a catalytic domain and two C-terminal domains. The enzyme is recombinantly expressed as an active approximately 56 kDa peptidase and includes both C terminal domains. Purified enzyme is converted to the approximately 34 kDa form by autolytic cleavage when incubated at 50 degrees C for 30 min, but retains full activity. In the present work, the Serratia peptidase (SPRK) is compared with the family representative proteinase K (PRK) from Tritirachium album Limber. Both enzymes show a relatively high thermal stability and a broad pH stability profile. SPRK possess superior stability towards SDS at 50 degrees C compared to PRK. On the other hand, SPRK is considerably more labile to removal of calcium ions. The activity profiles against temperature and pH differ for the two enzymes. SPRK shows both a broader pH optimum as well as a higher temperature optimum than PRK. Analysis of the catalytic properties of SPRK and PRK using the synthetic peptide succinyl-Ala-Ala-Pro-Phe-pNA as substrate showed that SPRK possesses a 3.5-4.5-fold higher kcat at the temperature range 12-37 degrees C, but a fivefold higher Km results in a slightly lower catalytic efficiency (kcat/Km) of SPRK compared to PRK. PMID- 16367748 TI - The 1.8 A crystal structure of a proteinase K-like enzyme from a psychrotroph Serratia species. AB - Proteins from organisms living in extreme conditions are of particular interest because of their potential for being templates for redesign of enzymes both in biotechnological and other industries. The crystal structure of a proteinase K like enzyme from a psychrotroph Serratia species has been solved to 1.8 A. The structure has been compared with the structures of proteinase K from Tritirachium album Limber and Vibrio sp. PA44 in order to reveal structural explanations for differences in biophysical properties. The Serratia peptidase shares around 40 and 64% identity with the Tritirachium and Vibrio peptidases, respectively. The fold of the three enzymes is essentially identical, with minor exceptions in surface loops. One calcium binding site is found in the Serratia peptidase, in contrast to the Tritirachium and Vibrio peptidases which have two and three, respectively. A disulfide bridge close to the S2 site in the Serratia and Vibrio peptidases, an extensive hydrogen bond network in a tight loop close to the substrate binding site in the Serratia peptidase and different amino acid sequences in the S4 sites are expected to cause different substrate specificity in the three enzymes. The more negative surface potential of the Serratia peptidase, along with a disulfide bridge close to the S2 binding site of a substrate, is also expected to contribute to the overall lower binding affinity observed for the Serratia peptidase. Clear electron density for a tripeptide, probably a proteolysis product, was found in the S' sites of the substrate binding cleft. PMID- 16367749 TI - Antifungal effects and mechanism of action of viscotoxin A3. AB - Viscotoxins are cationic proteins, isolated from different mistletoe species, that belong to the group of thionins, a group of basic cysteine-rich peptides of approximately 5 kDa. They have been shown to be cytotoxic to different types of cell, including animal, bacterial and fungal. The aim of this study was to obtain information on the cell targets and the mechanism of action of viscotoxin isoform A3 (VtA3). We describe a detailed study of viscotoxin interaction with fungal derived model membranes, its location inside spores of Fusarium solani, as well as their induced spore death. We show that VtA3 induces the appearance of ion channel-like activity, the generation of H2O2, and an increase in cytoplasmic free Ca2+. Moreover, we show that Ca2+ is involved in VtA3-induced spore death and increased H2O2 concentration. The data presented here strongly support the notion that the antifungal activity of VtA3 is due to membrane binding and channel formation, leading to destabilization and disruption of the plasma membrane, thereby supporting a direct role for viscotoxins in the plant defence mechanism. PMID- 16367750 TI - Optimal observability of sustained stochastic competitive inhibition oscillations at organellar volumes. AB - When molecules are present in small numbers, such as is frequently the case in cells, the usual assumptions leading to differential rate equations are invalid and it is necessary to use a stochastic description which takes into account the randomness of reactive encounters in solution. We display a very simple biochemical model, ordinary competitive inhibition with substrate inflow, which is only capable of damped oscillations in the deterministic mass-action rate equation limit, but which displays sustained oscillations in stochastic simulations. We define an observability parameter, which is essentially just the ratio of the amplitude of the oscillations to the mean value of the concentration. A maximum in the observability is seen as the volume is varied, a phenomenon we name system-size observability resonance by analogy with other types of stochastic resonance. For the parameters of this study, the maximum in the observability occurs at volumes similar to those of bacterial cells or of eukaryotic organelles. PMID- 16367751 TI - Parvalbumin deficiency in fast-twitch muscles leads to increased 'slow-twitch type' mitochondria, but does not affect the expression of fiber specific proteins. AB - Parvalbumin (PV), a small cytosolic protein belonging to the family of EF-hand calcium-binding proteins, is highly expressed in mammalian fast-twitch muscle fibers. By acting as a 'slow-onset' Ca2+ buffer, PV does not affect the rapid contraction phase, but significantly contributes to increase the rate of relaxation, as demonstrated in PV-/- mice. Unexpectedly, PV-/- fast-twitch muscles were considerably more resistant to fatigue than the wild-type fast twitch muscles. This effect was attributed mainly to the increased fractional volume of mitochondria in PV-/- fast-twitch muscle, extensor digitorum longus, similar to levels observed in the slow-twitch muscle, soleus. Quantitative analysis of selected mitochondrial proteins, mitochondrial DNA-encoded cytochrome oxidase c subunit I and nuclear DNA-encoded cytochrome oxidase c subunit Vb and F1-ATPase subunit beta revealed the PV-/- tibialis anterior mitochondria composition to be almost identical to that in wild-type soleus, but not in wild type fast-twitch muscles. Northern and western blot analyses of the same proteins in different muscle types and in liver are indicative of a complex regulation, probably also at the post-transcriptional level. Besides the function in energy metabolism, mitochondria in both fast- and slow-twitch muscles act as temporary Ca2+ stores and are thus involved in the shaping of Ca2+ transients in these cells. Previously observed altered spatio-temporal aspects of Ca2+ transients in PV-/- muscles are sufficient to up-regulate mitochondria biogenesis through the probable involvement of both calcineurin- and Ca2+/calmodulin-dependent kinase II dependent pathways. We propose that 'slow-twitch type' mitochondria in PV-/- fast muscles are aimed to functionally replace the slow-onset buffer PV based on similar kinetic properties of Ca2+ removal. PMID- 16367752 TI - Dissociation/association properties of a dodecameric cyclomaltodextrinase. Effects of pH and salt concentration on the oligomeric state. AB - As an effort to elucidate the quaternary structure of cyclomaltodextrinase I-5 (CDase I-5) as a function of pH and salt concentration, the dissociation/association processes of the enzyme were investigated under various pH and salt conditions. Previous crystallographic analysis of CDase I-5 indicated that it existed exclusively as a dodecamer at pH 7.0, forming an assembly of six 3D domain-swapped dimeric subunits. In the present study, analytical ultracentrifugation analysis suggested that CDase I-5 was present as a dimer in the pH range of 5.0-6.0, while the dodecameric form was predominant at pH values above 6.5. No dissociation of the dodecamer was observed at pH 7.0 and the above. Gel filtration chromatography showed that CDase I-5 dissociated into dimers at a rate of 8.58 x 10(-2) h(-1) at pH 6.0. A mutant enzyme with three histidine residues (H49, H89, and H539) substituted with valines dissociated into dimers faster than the wild-type enzyme at both pH 6.0 and 7.0. The tertiary structure indicated that the effect of pH on dissociation of the oligomer was mainly due to the protonation of H539. Unlike the pH-dependent process, the dissociation of wild-type CDase I-5 proceeded very fast at pH 7.0 in the presence of 0.2-1.0 M of KCl. Stopped-flow spectrophotometric analysis at various concentrations of KCl showed that the rate constants of dissociation (kd) from dodecamers into dimers were 5.96 s(-1) and 7.99 s(-1) in the presence of 0.2 M and 1.0 M of KCl, respectively. PMID- 16367753 TI - The three desulfoglucosinolate sulfotransferase proteins in Arabidopsis have different substrate specificities and are differentially expressed. AB - Sulfotransferases (SOTs) catalyse the transfer of a sulfate group from 3' phosphoadenosine 5'-phosphosulfate (PAPS) to an appropriate hydroxy group of various substrates with the parallel formation of 3'-phosphoadenosine 5' phosphate. In Arabidopsis thaliana, 18 SOT proteins (AtSOT) have been identified. Three of them, AtSOT16, AtSOT17 and AtSOT18, catalyse the sulfation of desulfoglucosinolates. The proteins were expressed in Escherichia coli, purified by affinity chromatography and used for enzyme kinetic studies. By establishing two types of enzyme assay using both 35S-labelled and unlabelled PAPS, separation of the products by HPLC, and detection of the products by monitoring radioactivity or UV absorption, the substrate specificities of the three AtSOT proteins were determined. They show different maximum velocities with several desulfoglucosinolates as substrates and differ in their affinity for desulfobenzylglucosinolate and PAPS. The sequences encoding AtSOT18 were amplified from Arabidopsis ecotypes C24 and Col0; the two expressed proteins differ in two out of 350 amino acids. These amino-acid variations led to different substrate specificities. Exchange of one of the two amino acids in AtSOT18 from C24 to the respective amino acid in AtSOT18 from Col0 gave the C24 protein the same substrate specificity as the wild-type AtSOT18 protein from Col0. All three desulfoglucosinolate AtSOT proteins are localized in the cytoplasm, as demonstrated by transient expression of fusion constructs with the green fluorescent protein in Arabidopsis protoplasts. Northern blot analysis indicated differential expression of the three AtSOT genes in plant organs and tissues at different developmental stages and during a light/darkness cycle. High (500 microM) and low (50 microM) sulfate concentrations in the medium did not influence the levels of expression. PMID- 16367754 TI - Occurrence of glucosylsucrose [alpha-D-glucopyranosyl- (1-->2)-alpha-D glucopyranosyl-(1-->2)-beta-D-fructofuranoside] and glucosylated homologues in cyanobacteria. Structural properties, cellular contents and possible function as thermoprotectants. AB - Little is known about the structure and function of oligosaccharides in cyanobacteria. In this study, a new class of saccharides from Nostoc was identified by MS and NMR techniques, consisting of alpha-D-glucopyranosyl-(1-->2) [alpha-D-glucopyranosyl-(1-->2)]n-beta-D-fructofuranosides ranging from the trisaccharide (n = 1) to decasaccharide (n = 8). In Nostoc ellipsosporum the cell content of saccharides increased 10-20-fold after heat stress (1 day, 40 degrees C) or during prolonged cultivation. Under these conditions the abundance of homologues of higher molecular mass (> pentasaccharide) increased and finally exceeded that of homologues of lower molecular mass including sucrose. Total intracellular content of the saccharides after heat stress was 5-10 mg x (g dry weight)(-1) corresponding to intracellular concentrations of 0.25-0.5% (w/v). A possible role of the oligosaccharides identified is in the protection of enzymes against heat inactivation. Whereas amylase from Nostoc was only weakly protected by the decasaccharide, alpha-amylase from porcine pancreas was more efficiently stabilized by the octasaccharide and decasaccharide. Evidence is presented for the widespread occurrence of the newly identified saccharides in cyanobacteria. The results are discussed including previous reports on cyanobacterial oligosaccharides and with respect to possible functions of these compounds in the living cell. PMID- 16367755 TI - Structure of amyloid beta fragments in aqueous environments. AB - Conformational studies on amyloid beta peptide (Abeta) in aqueous solution are complicated by its tendency to aggregate. In this study, we determined the atomic level structure of Abeta(28-42) in an aqueous environment. We fused fragments of Abeta, residues 10-24 (Abeta(10-24)) or 28-42 (Abeta(28-42)), to three positions in the C-terminal region of ribonuclease HII from a hyperthermophile, Thermococcus kodakaraensis (Tk-RNase HII). We then examined the structural properties in an aqueous environment. The host protein, Tk-RNase HII, is highly stable and the C-terminal region has relatively little interaction with other parts. CD spectroscopy and thermal denaturation experiments demonstrated that the guest amyloidogenic sequences did not affect the overall structure of the Tk RNase HII. Crystal structure analysis of Tk-RNase HII(1-197)-Abeta(28-42) revealed that Abeta(28-42) forms a beta conformation, whereas the original structure in Tk-RNase HII(1-213) was alpha helix, suggesting beta-structure formation of Abeta(28-42) within full-length Abeta in aqueous solution. Abeta(28 42) enhanced aggregation of the host protein more strongly than Abeta(10-24). These results and other reports suggest that after proteolytic cleavage, the C terminal region of Abeta adopts a beta conformation in an aqueous environment and induces aggregation, and that the central region of Abeta plays a critical role in fibril formation. This study also indicates that this fusion technique is useful for obtaining structural information with atomic resolution for amyloidogenic peptides in aqueous environments. PMID- 16367756 TI - The natural mutation by deletion of Lys9 in the thrombin A-chain affects the pKa value of catalytic residues, the overall enzyme's stability and conformational transitions linked to Na+ binding. AB - The catalytic competence of the natural thrombin mutant with deletion of the Lys9 residue in the A-chain (deltaK9) was found to be severely impaired, most likely due to modification of the 60-loop conformation and catalytic triad geometry, as supported by long molecular dynamics (MD) simulations in explicit water solvent. In this study, the pH dependence of the catalytic activity and binding of the low molecular mass inhibitor N-alpha-(2-naphthylsulfonyl-glycyl)-4 amidinophenylalanine-piperidine (alpha-NAPAP) to the wild-type (WT) and deltaK9 thrombin forms were investigated, along with their overall structural stabilities and conformational properties. Two ionizable groups were found to similarly affect the activity of both thrombins. The pKa value of the first ionizable group, assigned to the catalytic His57 residue, was found to be 7.5 and 6.9 in ligand-free deltaK9 and WT thrombin, respectively. Urea-induced denaturation studies showed higher instability of the deltaK9 mutant compared with WT thrombin, and disulfide scrambling experiments proved weakening of the interchain interactions, causing faster release of the reduced A-chain in the mutant enzyme. The sodium ion binding affinity was not significantly perturbed by Lys9 deletion, although the linked increase in intrinsic fluorescence was lower in the mutant. Essential dynamics (ED) analysis highlighted different conformational properties of the two thrombins in agreement with the experimental conformational stability data. Globally, these findings enhanced our understanding of the perturbations triggered by Lys9 deletion, which reduces the overall stability of the molecule, weakens the A-B interchain interactions, and allosterically perturbs the geometry and protonation state of catalytic residues of the enzyme. PMID- 16367757 TI - Induction of PPARbeta and prostacyclin (PGI2) synthesis by Raf signaling: failure of PGI2 to activate PPARbeta. AB - A role for the nuclear receptor peroxisome proliferator-activated receptor-beta (PPARbeta) in oncogenesis has been suggested by a number of observations but its precise role remains elusive. Prostaglandin I2 (PGI2, prostacyclin), a major arachidonic acid (AA) derived cyclooxygenase (Cox) product, has been proposed as a PPARbeta agonist. Here, we show that the 4-hydroxytamoxifen (4-OHT) mediated activation of a C-Raf-estrogen receptor fusion protein leads to the induction of both the PPARbeta and Cox-2 genes, concomitant with a dramatic increase in PGI2 synthesis. Surprisingly, however, 4-OHT failed to activate PPARbeta transcriptional activity, indicating that PGI2 is insufficient for PPARbeta activation. In agreement with this conclusion, the overexpression of ectopic Cox 2 and PGI2 synthase (PGIS) resulted in massive PGI2 synthesis but did not activate the transcriptional activity of PPARbeta. Conversely, inhibition of PGIS blocked PGI2 synthesis but did not affect the AA mediated activation of PPARbeta. Our data obtained with four different cell types and different experimental strategies do not support the prevailing opinion that PGI2 plays a significant role in the regulation of PPARbeta. PMID- 16367758 TI - Reactivity of the heme-dioxygen complex of the inducible nitric oxide synthase in the presence of alternative substrates. AB - Single turnover reactions of the inducible nitric oxide synthase oxygenase domain (iNOSoxy) in the presence of several non alpha-amino acid N-hydroxyguanidines and guanidines were studied by stopped-flow visible spectroscopy, and compared with reactions using the native substrates L-arginine (L-arg) or N(omega)-hydroxy-L arginine (NOHA). In experiments containing dihydrobiopterin, a catalytically incompetent pterin, and each of the studied substrates, L-arg, butylguanidine (BuGua), para-fluorophenylguanidine (FPhGua), NOHA, N-butyl- and N-(para fluorophenyl)-N'-hydroxyguanidines (BuNOHG and FPhNOHG), the formation of a iron(II) heme-dioxygen intermediate (Fe(II)O2) was always observed. The Fe(II)O2 species then decayed to iron(III) iNOSoxy at rates that were dependent on the nature of the substrate. Identical reactions containing the catalytically competent cofactor tetrahydrobiopterin (BH4), iNOSoxy and the three N hydroxyguanidines, all exhibited an initial formation of an Fe(II)O2 species that was successively converted to an Fe(III)NO complex and eventually to high-spin iron(III) iNOSoxy. The formation and decay kinetics of the Fe(III)NO complex did not vary greatly as a function of the N-hydroxyguanidine structure, but the formation of Fe(III)NO was substoichiometric in the cases of BuNOHG and FPhNOHG. Reactions between BH4-containing iNOSoxy and BuGua exhibited kinetics similar to those of the corresponding reaction with L-arginine, with formation of an Fe(II)O2 intermediate that was directly converted to high-spin iron(III) iNOSoxy. In contrast, no Fe(II)O2 intermediate was observed in the reaction of BH4 containing iNOSoxy and FPhGua. Multi-turnover reaction of iNOS with FPhGua did not lead to formation of NO or to hydroxylation of the substrate, contrary to reactions with BuGua or L-arg. Our results reveal how different structural and chemical properties of NOS substrate analogues can impact on the kinetics and reactivity of the Fe(II)O2 intermediate, and support an important role for substrate pKa during NOS oxygen activation. PMID- 16367759 TI - Identification of yeast aspartyl aminopeptidase gene by purifying and characterizing its product from yeast cells. AB - Aspartyl aminopeptidase (EC 3.4.11.21) cleaves only unblocked N-terminal acidic amino-acid residues. To date, it has been found only in mammals. We report here that aspartyl aminopeptidase activity is present in yeast. Yeast aminopeptidase is encoded by an uncharacterized gene in chromosome VIII (YHR113W, Saccharomyces Genome Database). Yeast aspartyl aminopeptidase preferentially cleaved the unblocked N-terminal acidic amino-acid residue of peptides; the optimum pH for this activity was within the neutral range. The metalloproteases inhibitors EDTA and 1.10-phenanthroline both inhibited the activity of the enzyme, whereas bestatin, an inhibitor of most aminopeptidases, did not affect enzyme activity. Gel filtration chromatography revealed that the molecular mass of the native form of yeast aspartyl aminopeptidase is approximately 680,000. SDS/PAGE of purified yeast aspartyl aminopeptidase produced a single 56-kDa band, indicating that this enzyme comprises 12 identical subunits. PMID- 16367760 TI - DNA mediated disassembly of hRad51 and hRad52 proteins and recruitment of hRad51 to ssDNA by hRad52. AB - Purified human Rad51 and Rad52 proteins exhibit multiple oligomeric states, in vitro. Single-stranded DNA (ssDNA) renders high molecular weight aggregates of both proteins into smaller and soluble forms that include even the monomers. Consequently, these proteins that have a propensity to interact with each other's higher order forms by themselves, start interacting with monomeric forms in the presence of ssDNA, presumably reflecting the steps of protein assembly on DNA. In the same conditions, DNA binding assays reveal hRad52-mediated recruitment of hRad51 on ssDNA. Put together, these studies hint at DNA-induced disassembly of higher-order forms of Rad51 and Rad52 proteins as steps that precede protein assembly during hRad51 presynapsis on DNA, in vitro. PMID- 16367761 TI - Active site residues governing substrate selectivity and polyketide chain length in aloesone synthase. AB - Aloesone synthase (ALS) and chalcone synthase (CHS) are plant-specific type III poyketide synthases sharing 62% amino acid sequence identity. ALS selects acetyl CoA as a starter and carries out six successive condensations with malonyl-CoA to produce a heptaketide aloesone, whereas CHS catalyses condensations of 4 coumaroyl-CoA with three malonyl-CoAs to generate chalcone. In ALS, CHS's Thr197, Gly256, and Ser338, the active site residues lining the initiation/elongation cavity, are uniquely replaced with Ala, Leu, and Thr, respectively. A homology model predicted that the active site architecture of ALS combines a 'horizontally restricting' G256L substitution with a 'downward expanding' T197A replacement relative to CHS. Moreover, ALS has an additional buried pocket that extends into the 'floor' of the active site cavity. The steric modulation thus facilitates ALS to utilize the smaller acetyl-CoA starter while providing adequate volume for the additional polyketide chain extensions. In fact, it was demonstrated that CHS like point mutations at these positions (A197T, L256G, and T338S) completely abolished the heptaketide producing activity. Instead, A197T mutant yielded a pentaketide, 2,7-dihydroxy-5-methylchromone, while L256G and T338S just afforded a triketide, triacetic acid lactone. In contrast, L256G accepted 4-coumaroyl-CoA as starter to efficiently produce a tetraketide, 4-coumaroyltriacetic acid lactone. These results suggested that Gly256 determines starter substrate selectivity, while Thr197 located at the entrance of the buried pocket controls polyketide chain length. Finally, Ser338 in proximity of the catalytic Cys164 guides the linear polyketide intermediate to extend into the pocket, thus leading to formation of the hepataketide in Rheum palmatum ALS. PMID- 16367762 TI - Characterization of rat cathepsin E and mutants with changed active-site residues and lacking propeptides and N-glycosylation, expressed in human embryonic kidney 293T cells. AB - To study the roles of the catalytic activity, propeptide, and N-glycosylation of the intracellular aspartic proteinase cathepsin E in biosynthesis, processing, and intracellular trafficking, we constructed various rat cathepsin E mutants in which active-site Asp residues were changed to Ala or which lacked propeptides and N-glycosylation. Wild-type cathepsin E expressed in human embryonic kidney 293T cells was mainly found in the LAMP-1-positive endosomal organelles, as determined by immunofluorescence microscopy. Consistently, pulse-chase analysis revealed that the initially synthesized pro-cathepsin E was processed to the mature enzyme within a 24 h chase. This process was completely inhibited by brefeldin A and bafilomycin A, indicating its transport from the endoplasmic reticulum (ER) to the endosomal acidic compartment. Mutants with Asp residues in the two active-site consensus motifs changed to Ala and lacking the propeptide (Leu23-Phe58) and the putative ER-retention sequence (Ser59-Asp98) were neither processed nor transported to the endosomal compartment. The mutant lacking the ER retention sequence was rapidly degraded in the ER, indicating the importance of this sequence in correct folding. The single (N92Q or N324D) and double (N92Q/N324D) N-glycosylation-deficient mutants were neither processed into a mature form nor transported to the endosomal compartment, but were stably retained in the ER without degradation. These data indicate that the catalytic activity, propeptides, and N-glycosylation of this protein are all essential for its processing, maturation, and trafficking. PMID- 16367763 TI - Recombinant hemoglobin betaG83C-F41Y. AB - We have engineered a stable octameric hemoglobin (Hb) of molecular mass 129 kDa, a dimer of recombinant hemoglobin (rHb betaG83C-F41Y) tetramers joined by disulfide bonds at the beta83 position. One of the major problems with oxygen carriers based on acellular hemoglobin solutions is vasoactivity, a limitation which may be overcome by increasing the molecular size of the carrier. The oxygen equilibrium curves showed that the octameric rHb betaG83C-F41Y exhibited an increased oxygen affinity and a decreased cooperativity. The CO rebinding kinetics, auto-oxidation kinetics, and size exclusion chromatography did not show the usual dependence on protein concentration, indicating that this octamer was stable and did not dissociate easily into tetramers or dimers at low concentration. These results were corroborated by the experiments with haptoglobin showing no interaction between octameric rHb betaG83C-F41Y and haptoglobin, a plasma glycoprotein that binds the Hb dimers and permits their elimination from blood circulation. The lack of dimers could be explained if there are two disulfide bridges per octamer, which would be in agreement with the lack of reactivity of the additional cysteine residues. The kinetics of reduction of the disulfide bridge by reduced glutathione showed a rate of 1000 M(-1) x h( 1) (observed time coefficient of 1 h at 1 mM glutathione) at 25 degrees C. Under air, the cysteines are oxidized and the disulfide bridge forms spontaneously; the kinetics of the tetramer to octamer reaction displayed a bimolecular reaction of time coefficient of 2 h at 11 microM Hb and 25 degrees C. In addition, the octameric rHb betaG83C-F41Y was resistant to potential reducing agents present in fresh plasma. PMID- 16367764 TI - Identification of a cis-acting element required for dendritic targeting of activity-regulated cytoskeleton-associated protein mRNA. AB - The mRNA encoding activity-regulated cytoskeleton-associated protein (Arc) is known to be targeted to dendritic regions that have received strong synaptic inputs. However, the cis-acting elements in Arc mRNA that mediate dendritic targeting have not been identified. To identify the dendritic targeting element (DTE) in rat Arc mRNA, we expressed reporter mRNAs containing various regions of Arc in primary hippocampal neurones and analysed their subcellular distribution by in situ hybridization. Here, we report that the 3'-untranslated region of rat Arc mRNA contains a 350-nucleotide DTE with strong dendritic targeting activity and another 370-nucleotide sequence with weaker dendritic targeting activity. The 350-nucleotide DTE does not share any obvious sequence similarity with other known DTEs previously reported. PMID- 16367765 TI - Bi-directional regulation of postsynaptic cortactin distribution by BDNF and NMDA receptor activity. AB - Abstract Cortactin is an F-actin-associated protein which interacts with the postsynaptic scaffolding protein Shank at the SH3 domain and is localized within the dendritic spine in the mouse neuron. Green fluorescent protein (GFP)-based time-lapse imaging revealed cortactin redistribution from dendritic cytoplasm to postsynaptic sites by application of brain-derived neurotrophic factor (BDNF). This response was mediated by mitogen-activated protein (MAP) kinase activation and was dependent on the C-terminal SH3 domain. In contrast, activation of N methyl-D-aspartate (NMDA) receptors induced loss of cortactin from postsynaptic sites. This NMDA-dependent redistribution was blocked by an Src family kinase inhibitor. Conversely, increasing Src family kinase activity induced cortactin phosphorylation and loss of cortactin from the postsynaptic sites. Finally, blocking of endogenous BDNF reduced the amount of cortactin at the postsynaptic sites and an NMDA receptor antagonist prevented this reduction. These results indicate the importance of counterbalance between BDNF and NMDA receptor-mediated signalling in the reorganization of the postsynaptic actin cytoskeleton during neuronal development. PMID- 16367766 TI - Transient decrease in F-actin may be necessary for translocation of proteins into dendritic spines. AB - It remains poorly understood as to how newly synthesized proteins that are required to act at specific synapses are translocated into only selected subsets of potentiated dendritic spines. Here, we report that F-actin, a major component of the skeletal structure of dendritic spines, may contribute to the regulation of synaptic specificity of protein translocation. We found that the stabilization of F-actin blocked the translocation of GFP-CaMKII and inhibited the diffusion of 3-kDa dextran into spines (in 2-3 weeks cultures). Neuronal activation in hippocampal slices and cultured neurons led to an increase in the activation (decrease in the phosphorylation) of the actin depolymerization factor, cofilin, and a decrease in F-actin. Furthermore, the induction of long-term potentiation by tetanic stimulation induced local transient depolymerization of F-actin both in vivo and in hippocampal slices (8-10 weeks), and this local F-actin depolymerization was blocked by APV, a N-methyl-D-aspartate (NMDA) receptor antagonist. These results suggest that F-actin may play a role in synaptic specificity by allowing protein translocation into only potentiated spines, gated through its depolymerization, which is probably triggered by the activation of NMDA receptors. PMID- 16367768 TI - Differential expression of calcineurin A subunit mRNA isoforms during rat hippocampal and cerebellar development. AB - Calcineurin (protein phosphatase 2B) is a calcium-dependent serine-threonine phosphatase. It has diverse roles and is centrally involved in synaptic plasticity. The catalytic A subunit of calcineurin has three isoforms, alpha, beta and gamma. Their expression and ontogeny in the brain has not been systematically investigated; such data become important with a report that PPP3CC, the gene encoding calcineurin Agamma, is a susceptibility gene for schizophrenia, and the finding that its expression is decreased in the disorder. We used in situ hybridization histochemistry to measure the relative transcript abundance of calcineurin Agamma and the other catalytic isoforms, Aalpha and Abeta, during development of the Sprague-Dawley rat hippocampus and cerebellum. All three isoforms are present in both regions at all time points [embryonic day 19 (E19) to postnatal day 42 (P42)] and undergo developmental regulation, but differ in their ontogenic profile. Calcineurin Aalpha and Abeta mRNAs increased from E19 through to adulthood, whereas Agamma mRNA was most highly expressed during early developmental stages. Calcineurin Aalpha and Abeta mRNAs positively correlated with synaptophysin mRNA (a synaptic marker), whilst Agamma mRNA was either unrelated to, or negatively correlated, with this transcript. These data confirm that all three calcineurin A subunits are expressed in the rodent brain, and indicate that calcineurin Agamma may have different roles than Aalpha and Abeta. The data also suggest a potential importance of calcineurin Agamma in neurodevelopment, and in the genetically influenced neurodevelopmental disturbance that is thought to underlie schizophrenia. PMID- 16367767 TI - Local activation of the nitric oxide/cyclic guanosine monophosphate pathway in growth cones regulates filopodial length via protein kinase G, cyclic ADP ribose and intracellular Ca2+ release. AB - Nitric oxide (NO) is a gaseous messenger that has been shown to affect growth cone motility and neurite outgrowth in several model systems, but how NO brings about its effects is not understood. We have previously demonstrated that global and long-term application of NO to Helisoma trivolvis B5 neurons results in a transient increase in filopodial length, decrease in filopodial number and decrease in neurite outgrowth, all of which are mediated via soluble guanylyl cyclase (sGC) and involve an increase in the intracellular Ca2+ concentration [S. Van Wagenen & V. Rehder (1999)Journal of Neurobiology, 39, 168-185; K.R. Trimm & V. Rehder (2004) European Journal of Neuroscience, 19, 809-818]. The goal of the current study was twofold: to investigate the effects of short-term NO exposure on individual growth cones and to further elucidate the downstream pathway through which NO exerts its effects. Local application of the NO donor NOC-7 for 10-20 ms via puffer micropipette resulted in a transient increase in filopodial length and a small decrease in filopodial number. We show evidence that these effects of NO are mediated via sGC, protein kinase G and cyclic ADP ribose, resulting in the release of Ca2+ from intracellular stores, probably of the ryanodine-sensitive type. These results suggest that growth cones expressing sGC are highly sensitive to local and short-term exposure to NO, which they may experience during pathfinding, and that the stereotyped response of transient filopodial elongation seen in B5 neurons in response to NO requires intracellular Ca2+ release. PMID- 16367769 TI - An investigation into the potential for activity-dependent regeneration of the rubrospinal tract after spinal cord injury. AB - We tested whether regeneration of transected rubrospinal tract (RST) axons is facilitated by a prolonged electrical stimulation of these axons. A peripheral nerve was grafted to the transected RST at the cervical level (C4/5) of adult rats, providing a permissive environment for regeneration of rubrospinal axons. Direct antidromic stimulation of the RST was applied immediately after grafting through a microwire inserted just rostral to the RST lesion, using a 1-h 20-Hz supramaximal stimulation protocol. Stimulation caused no direct damage to rubrospinal axons, and was sufficient to recruit the entire rubrospinal tract. In control animals that had a nerve graft and implanted microwire with no stimulation, there were 42.7 +/- 10.2 rubrospinal neurons regenerated into the graft at 8 weeks, as assessed by retrograde labelling. In test animals that were stimulated there were 28.2 +/- 7.4 back-labelled neurons, not significantly different from control, indicating that this stimulation did not improve the regenerative capacity of rubrospinal neurons. Furthermore, reverse-transcriptase polymerase chain reaction and in situ hybridization for brain-derived neurotrophic factor (BDNF) and/or growth-associated protein-43 (GAP-43) expression in rubrospinal neurons revealed no significant difference between stimulated and unstimulated groups at 48 h after injury, with either 1 or 8 h of stimulation. In summary, direct stimulation of the injured RST axons for the periods tested does not increase expression of GAP-43 and BDNF, and ultimately does not promote regeneration of these central nervous system axons. PMID- 16367770 TI - Chondroitinase ABC combined with neural stem/progenitor cell transplantation enhances graft cell migration and outgrowth of growth-associated protein-43 positive fibers after rat spinal cord injury. AB - We previously reported that the transplantation of neural stem/progenitor cells (NSPCs) can contribute to the repair of injured spinal cord in adult rats and monkeys. In some cases, however, most of the transplanted cells adhered to the cavity wall and failed to migrate and integrate into the host spinal cord. In this study we focused on chondroitin sulfate proteoglycan (CSPG), a known constituent of glial scars that is strongly expressed after spinal cord injury (SCI), as a putative inhibitor of NSPC migration in vivo. We hypothesized that the digestion of CSPG by chondroitinase ABC (C-ABC) might promote the migration of transplanted cells and neurite outgrowth after SCI. An in vitro study revealed that the migration of NSPC-derived cells was inhibited by CSPG and that this inhibitory effect was attenuated by C-ABC pre-treatment. Consistently, an in vivo study of C-ABC treatment combined with NSPC transplantation into injured spinal cord revealed that C-ABC pre-treatment promoted the migration of the transplanted cells, whereas CSPG-immunopositive scar tissue around the lesion cavity prevented their migration into the host spinal cord in the absence of C-ABC pre-treatment. Furthermore, this combined treatment significantly induced the outgrowth of a greater number of growth-associated protein-43-positive fibers at the lesion epicentre, compared with NSPC transplantation alone. These findings suggested that the application of C-ABC enhanced the benefits of NSPC transplantation for SCI by reducing the inhibitory effects of the glial scar, indicating that this combined treatment may be a promising strategy for the regeneration of injured spinal cord. PMID- 16367771 TI - Suppression of fibrous scarring in spinal cord injury of rat promotes long distance regeneration of corticospinal tract axons, rescue of primary motoneurons in somatosensory cortex and significant functional recovery. AB - Traumatic injury of the central nervous system results in formation of a collagenous basement membrane-rich fibrous scar in the lesion centre. Due to accumulation of numerous axon-growth inhibitory molecules the lesion scar is considered a major impediment for axon regeneration. Following transection of the dorsal corticospinal tract (CST) at thoracic level 8 in adult rats, transient suppression of collagenous scarring in the lesion zone by local application of a potent iron chelator and cyclic adenosine monophosphate resulted in the delay of fibrous scarring. Treated animals displayed long-distance growth of CST axons through the lesion area extending for up to 1.5-2 cm into the distal cord. In addition, the treatment showed a strong neuroprotective effect, rescuing cortical motoneurons projecting into the CST that normally die (30%) after thoracic axotomy. Further, anterogradely traced CST axons regenerated through both grey and white matter and developed terminal arborizations in grey matter regions. In contrast to controls, injured animals receiving treatment showed significant functional recovery in the open field, in the horizontal ladder and in CatWalk locomotor tasks. We conclude that the fibrous lesion scar plays a pivotal role as a growth barrier for regenerating axons in adult spinal cord and that a delay in fibrotic scarring by local inhibition of collagen biosynthesis and basement membrane deposition is a promising and unique therapeutic strategy for treating human spinal trauma. PMID- 16367772 TI - Nestin-expressing cells and their relationship to mitotically active cells in the subventricular zones of the adult rat. AB - Nestin is a protein that is thought to be expressed in neural stem cells; however, there is a paucity of data on nestin expression in vivo, and little is known of the relationship between nestin and mitotically active cell populations in the subventricular zones (SVZ). In this study, the subventricular zone of the third ventricle contained a high proportion of cells that expressed nestin, while there were significantly fewer cells that expressed nestin in the SVZ of the lateral ventricles. In contrast, bromodeoxyuridine (BrdU) immunoreactivity was the diametric opposite, being higher in the SVZ of the lateral ventricle than in the SVZ of the third ventricle. Morphological and anatomical evidence suggests that nestin-expressing cells in these two areas may be different cell types. In a separate set of experiments, an acute localized lesion was induced adjacent to one of the ventricles. While the number of BrdU cells and Ki-67 cells in the SVZs increased with this manipulation, the number of nestin-expressing cells did not change significantly. These data indicate that the expression of nestin does not correlate with mitotic activity in cells of the SVZs under either normal or inflammatory conditions. It is hypothesized that nestin-expressing cells in the SVZs may give way to transit amplifying cells that in turn give way to immature neurons or glia. These transit-amplifying cells may have a much higher rate of mitosis than nestin-positive cells and may react to neural damage by increasing their rate of proliferation. PMID- 16367773 TI - Late N-methyl-D-aspartate receptor blockade rescues hippocampal neurons from excitotoxic stress and death after 4-aminopyridine-induced epilepsy. AB - The intrahippocampal perfusion of 4-aminopyridine (4-AP) in the rat produces immediate seizures and delayed neuronal death, due to the overactivation of N methyl-D-aspartate (NMDA) receptors by endogenous glutamate released from nerve endings. With the same time course, 4-AP also induces the expression of the cell stress marker heat shock protein 70 (HSP70) in the contralateral non-damaged hippocampus. We have used this experimental model to study the mechanisms of the delayed neuronal stress and death. The NMDA receptor antagonist (+)-5-methyl 10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate (MK-801), administered intraperitoneally 30 or 60 but not 120 min after 4-AP perfusion, when animals show intense electroencephalography epileptiform activity, prevented the delayed neurodegeneration whereas the seizures continued for about 3 h as in the control animals. With an identical time window, MK-801 treatment also modified the pattern of HSP70 expression; the protein was expressed in the protected perfused hippocampus but no longer in the undamaged contralateral hippocampus. The possible role of Ca2+ in the delayed cell death and HSP70 expression was also studied by coperfusing the intracellular Ca2+ chelator 1,2 bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid tetrakis(acetoxymethyl ester) with 4-AP. This treatment resulted in protective and HSP70 effects very similar to those of MK-801. These results suggest that the seizures are not linked to neurodegeneration and that NMDA receptors need to be continuously overactivated by endogenous glutamate for at least 60 min in order to induce delayed neuronal stress and death, which are dependent on Ca2+ entry through the NMDA receptor channel. PMID- 16367774 TI - Differential responses to NMDA receptor activation in rat hippocampal interneurons and pyramidal cells may underlie enhanced pyramidal cell vulnerability. AB - Hippocampal interneurons are generally more resistant than pyramidal cells to excitotoxic insults. Because NMDA receptors play a crucial role in neurodegeneration, we have compared the response to exogenous NMDA in CA1 pyramidal cells and interneurons of the stratum oriens using combined whole-cell patch-clamp recording and ratiometric Ca2+ imaging. In voltage-clamp, current clamp or in nominally Mg2+-free medium, NMDA (10 microM; 3-5 min exposure in the presence of tetrodotoxin) induced a markedly larger inward current and Ca2+ rise in pyramidal cells than in interneurons. Pyramidal cells also showed a more pronounced voltage dependence in their response to NMDA. We hypothesized that this enhanced response to NMDA receptor activation in pyramidal cells could underlie their increased vulnerability to excitotoxicity. Using loss of dye as an indicator of degenerative membrane disruption, interneurons tolerated continuous exposure to a high concentration of NMDA (30 microM) for longer periods than pyramidal cells. This acute neurodegeneration in pyramidal cells was independent of intracellular Ca2+, because high intracellular BAPTA (20 mM) did not prolong survival time. Thus, a plausible explanation for the enhanced sensitivity of pyramidal neurons to excitotoxic insults associated with cerebral ischemia is their greater response to NMDA receptor activation, which may reflect differences in NMDA receptor expression and/or subunit composition. PMID- 16367775 TI - Effects of M-current modulators on the excitability of immature rat spinal sensory and motor neurones. AB - M-currents have been shown to control neuronal excitability in a variety of central and peripheral neurones. Here we studied the effects of specific M current modulators on the excitability of spinal neurones and their response to synaptic activation. Experiments were performed in vitro using the hemisected spinal cord from 7- to 11-day-old rats. Intracellular recordings were obtained from lumbar deep dorsal horn and motor neurones. Neuronal excitability was assessed by applying outward current pulses and synaptic responses were elicited by activation of a lumbar dorsal root. The M-current antagonist 10,10-bis(4 pyridinylmethyl)-9(10H)-anthracenone (XE-991) and the agonist retigabine were superfused at 10 microM. Retigabine produced hyperpolarization and a large decrease in the excitability of motor (7/7) and dorsal horn neurones (11/12). The effects of retigabine were fully reversed by XE-991. XE-991 induced depolarization of most neurones tested and a large increase in the excitability of motor neurones (7/7) but only a weak increase in the excitability of a proportion of dorsal horn neurones (4/10). The effects of XE-991 were partly reversed by retigabine. Consistent with their effects on neuronal excitability, retigabine showed a general depressant effect on synaptic transmission, whereas XE-991 showed the opposite tendency to potentiate responses to dorsal root stimulation, particularly in motor neurones. The results show that retigabine can depress spinal excitability and the transmission of nociceptive information. Results also indicate a post-synaptic expression of functional M-currents in most motor neurones and a considerable proportion of deep dorsal horn neurones. PMID- 16367776 TI - Physiological characterization, localization and synaptic inputs of bursting and nonbursting neurons in the trigeminal principal sensory nucleus of the rat. AB - A population of neurons in the trigeminal principal sensory nucleus (NVsnpr) fire rhythmically during fictive mastication induced in the in vivo rabbit. To elucidate whether these neurons form part of the central pattern generator (CPG) for mastication, we performed intracellular recordings in brainstem slices taken from young rats. Two cell types were defined, nonbursting (63%) and bursting (37%). In response to membrane depolarization, bursting cells, which dominated in the dorsal part of the NVsnpr, fired an initial burst followed by single spikes or recurring bursts. Non-bursting neurons, scattered throughout the nucleus, fired single action potentials. Microstimulation applied to the trigeminal motor nucleus (NVmt), the reticular border zone surrounding the NVmt, the parvocellular reticular formation or the nucleus reticularis pontis caudalis (NPontc) elicited a postsynaptic potential in 81% of the neurons tested for synaptic inputs. Responses obtained were predominately excitatory and sensitive to glutamatergic antagonists DNQX and/or APV. Some inhibitory and biphasic responses were also evoked. Bicuculline methiodide or strychnine blocked the IPSPs indicating that they were mediated by GABA(A) or glycinergic receptors. About one-third of the stimulations activated both types of neurons antidromically, mostly from the masseteric motoneuron pool of NVmt and dorsal part of NPontc. In conclusion, our new findings show that some neurons in the dorsal NVsnpr display both firing properties and axonal connections which support the hypothesis that they may participate in masticatory pattern generation. Thus, the present data provide an extended basis for further studies on the organization of the masticatory CPG network. PMID- 16367777 TI - SK channels control the firing pattern of midbrain dopaminergic neurons in vivo. AB - A vast body of experimental in vitro work and modelling studies suggests that the firing pattern and/or rate of a majority of midbrain dopaminergic neurons may be controlled in part by Ca2+-activated K+ channels of the SK type. However, due to the lack of suitable tools, in vivo evidence is lacking. We have taken advantage of the development of the water-soluble, medium potency SK blocker N-methyl laudanosine (CH3-L) to test this hypothesis in anaesthetized rats. In the lateral ventral tegmental area, CH3-L iontophoresis onto dopaminergic neurons significantly increased the coefficient of variation of their interspike intervals and the percentage of spikes generated in bursts as compared to the control condition. The effect of CH3-L persisted in the presence of a specific GABA(A) antagonist, suggesting a direct effect. It was robust and reversible, and was also observed in the substantia nigra. Control experiments demonstrated that the effect of CH3-L could be entirely ascribed to its blockade of SK channels. On the other hand, the firing pattern of noradrenergic neurons was much less affected by CH3-L. We provide here the first demonstration of a major role of SK channels in the control of the switch between tonic and burst firing of dopaminergic neurons in physiological conditions. This study also suggests a new strategy to develop modulators of the dopaminergic (DA) system, which could be of interest in the treatment of Parkinson's disease, and perhaps other diseases in which DA pathways are dysfunctional. PMID- 16367778 TI - Effects of extracellular delta-aminolaevulinic acid on sodium currents in acutely isolated rat hippocampal CA1 neurons. AB - The effects of delta-aminolaevulinic acid (ALA) on voltage-gated sodium channel (VGSC) currents (I(Na)) in acutely isolated hippocampal CA1 neurons from 10- to 12-day-old Wistar rats were examined by using the whole-cell patch-clamp technique under voltage-clamp conditions. ALA from 0.01 microm to 20 microm was applied to the recorded neurons. Low concentrations of ALA (0.01-1.0 microM) increased I(Na) amplitude, whereas high concentrations of ALA (5.0-20.0 microM) decreased it. The average I(Na) amplitude reached a maximum of 117.4 +/- 3.9% (n = 9, P < 0.05) with 0.1 microM ALA, and decreased to 78.1 +/- 3.8% (n = 13, P < 0.05) with 10 microm ALA. ALA shifted the steady-state activation and inactivation curves of I(Na) in the hyperpolarizing direction with different V0.5, suggesting that ALA could depress the opening threshold of the voltage gated sodium channel (VGSC) and thus increase the excitability of neurons through facilitating the opening of VGSC. The time course of recovery from inactivation was significantly prolonged at both low and high concentrations of ALA, whereas either low or high concentrations of ALA had no significant effect on the attenuation of I(Na) during stimulation at 5 Hz, indicating that the effect of ALA on VGSC is state-independent. Furthermore, we found that application of ascorbic acid, which blocks pro-oxidative effects in neurons, could prevent the increase of I(Na) amplitude at low concentrations of ALA. Baclofen, an agonist of GABAb receptors, induced some similar effects to ALA on VGSC, whereas bicuculline, an antagonist of GABAa receptors, could not prevent ALA-induced effects on VGSC. These results suggested that ALA regulated VGSC mainly through its pro-oxidative effects and GABAb receptor-mediated effects. PMID- 16367779 TI - Dopamine regulates melanopsin mRNA expression in intrinsically photosensitive retinal ganglion cells. AB - In mammals a subpopulation of retinal ganglion cells are intrinsically photosensitive (ipRGCs), express the photopigment melanopsin, and play an important role in the regulation of the nonimage-forming visual system. We have recently reported that melanopsin mRNA and protein levels in the rat retina are under photic and circadian control. The aim of the present work was to investigate the mechanisms that control melanopsin expression in the rat retina. We discovered that dopamine (DA) is involved in the regulation of melanopsin mRNA, possibly via dopamine D2 receptors that are located on these ipRGCs. Interestingly, we also discovered that pituitary adenylate cyclase-activating peptide (PACAP) mRNA levels are affected by DA. Dopamine synthesis and release in the retina are regulated by the rod and the cone photoreceptors via retinal circuitry; our new data indicate that DA controls melanopsin expression, indicating that classical photoreceptors may modulate the transcription of this new photopigment. Our study also suggests that DA may have an important role in mediating the light signals that are used for circadian entrainment and for other responses that are mediated by the nonimage-forming visual system. PMID- 16367781 TI - Olfactory receptors on the cockroach antenna signal odour ON and odour OFF by excitation. AB - A morphologically identifiable type of olfactory sensillum on the antenna of the American cockroach contains a pair of ON and OFF cells that responds oppositely to changes in the concentration of fruit odours. The odour of lemon oil was used to study the accuracy with which these cells can discriminate between rapid step like, ramp-like and oscillating changes in odour concentration. The discharge rates of both cells are not only affected by the actual concentration at particular instants in time (instantaneous concentration) but also by the rate at which concentration changes. The impulse frequency of the fruit odour ON cell is high when odour concentration is high, but higher still when odour concentration is also rising. Conversely, the impulse frequency of the fruit odour OFF cell is high when odour concentration is low and higher still when odour concentration is also falling. Thus, the effect of odour concentration on the responses of both cells is reinforced by the rate of change. Sensitivity to the rate of concentration change becomes greater when the rate is low. Because of the high sensitivity to low rates of change, these cells are optimized to detect fluctuations in fruit odour concentration. Whereas the ON cell signals the arrival and presence of fruit odour, the OFF cell detects its termination and absence. These cells provide excitatory responses for both increase and decrease in fruit odour concentration and may therefore reinforce contrast information. PMID- 16367780 TI - Direct pyrogenic input from prostaglandin EP3 receptor-expressing preoptic neurons to the dorsomedial hypothalamus. AB - Fever is induced by a neuronal mechanism in the brain. Prostaglandin (PG) E2 acts as a pyrogenic mediator in the preoptic area (POA) probably through the EP3 subtype of PGE receptor expressed on GABAergic neurons, and this PGE2 action triggers neuronal pathways for sympathetic thermogenesis in peripheral effector organs including brown adipose tissue (BAT). To explore pyrogenic efferent pathways from the POA, we determined projection targets of EP3 receptor expressing POA neurons with a special focus on rat hypothalamic regions including the dorsomedial hypothalamic nucleus (DMH), which is known as a center for autonomic responses to stress. Among injections of cholera toxin b-subunit (CTb), a retrograde tracer, into hypothalamic regions at the rostrocaudal level of the DMH, injections into the DMH, lateral hypothalamic area (LH) and dorsal hypothalamic area (DH) resulted in EP3 receptor immunolabelling in substantial populations of CTb-labeled neurons in the POA. Bilateral microinjections of muscimol, a GABA(A) receptor agonist, into the DMH and a ventral region of the DH, but not those into the LH, inhibited thermogenic (BAT sympathetic nerve activity, BAT temperature, core body temperature and expired CO2) and cardiovascular (arterial pressure and heart rate) responses to an intra-POA PGE2 microinjection. Further immunohistochemical observations revealed a close association of POA-derived GABAergic axon swellings with DMH neurons projecting to the medullary raphe regions where sympathetic premotor neurons for febrile and thermoregulatory responses are localized. These results suggest that a direct projection of EP3 receptor-expressing POA neurons to the DMH/DH region mediates febrile responses via a GABAergic mechanism. PMID- 16367782 TI - Electrophysiological and behavioural characterization of gustatory responses to antennal 'bitter' taste in honeybees. AB - We combined behavioural and electrophysiological experiments to study whether bitter taste is perceived at the antennal level in honeybees, Apis mellifera. Our behavioural studies showed that neither quinine nor salicin delivered at one antenna at different concentrations induced a retraction of the proboscis once it was extended in response to 1 M sucrose solution delivered to the opposite antenna. Bees that extended massively their proboscis to 1 M sucrose responded only partially when stimulated with a mixture of 1 M sucrose and 100 mM quinine. The mixture of 1 m sucrose and 100 mM salicin had no such suppressive effect. No behavioural suppression was found for mixtures of salt solution and either bitter substance. Electrophysiological recordings of taste sensillae at the antennal tip revealed sensillae that responded specifically either to sucrose or salt solutions, but none responded to the bitter substances quinine and salicin at the different concentrations tested. The electrophysiological responses of sensillae to 15 mM sucrose solution were inhibited by a mixture of 15 mM sucrose and 0.1 mM quinine, but not by a mixture of 15 mM sucrose and 0.1 mM salicin. The responses of sensillae to 50 mM NaCl were reduced by a mixture of 50 mm NaCl and 1 mM quinine but not by a mixture of 50 mM NaCl and 1 mM salicin. We concluded that no receptor cells for the bitter substances tested, exist at the level of the antennal tip of the honeybee and that antennal bitter taste is not represented as a separate perceptual quality. PMID- 16367783 TI - Calcium concentration changes during sensory transduction in spider mechanoreceptor neurons. AB - Most mechanoreceptor neurons encode mechanical signals into action potential trains within the same cell. Evidence suggests that intracellular calcium ion concentration, [Ca2+], increases during mechanotransduction, either by direct entry through mechanically activated channels or indirectly through voltage activated calcium channels. However, little is known about the amounts of calcium involved or its roles in mechanotransduction. We estimated [Ca2+] in mechanoreceptor neurons of the spider, Cupiennius salei, during mechanical stimulation using Oregon Green BAPTA-1, and a single-compartment model of [Ca2+] as a function of action potential firing rate. Resting [Ca2+] was approximately 400 nM and increased to up to 2 microM at 30 action potentials/s. Similar levels of resting and stimulated [Ca2+] were obtained in the cell soma, axon and two parts of the sensory dendrite, including the region immediately adjacent to the site of sensory transduction. The time constant of rise and fall of [Ca2+] was 1 5 s in the dendrite and axon, but up to 15 s in the soma. Calcium elevation was dependent on action potentials and could not be induced by the receptor potential alone. Blockade of voltage-activated calcium channels by nickel ions prevented calcium increase, but thapsigargin, which empties intracellular calcium stores, had no effect. Estimates of calcium entry per action potential from fluorescence changes agreed approximately with estimates based on action potential voltage time profile and previous reports of calcium channel properties. This first report of calcium levels during transduction in spiking mechanoreceptors suggests that calcium signaling plays important roles in primary somatosensory neurons. PMID- 16367784 TI - A population-based model of the nonlinear dynamics of the thalamocortical feedback network displays intrinsic oscillations in the spindling (7-14 Hz) range. AB - The thalamocortical network is modelled using the Wilson-Cowan equations for neuronal population activity. We show that this population model with biologically derived parameters possesses intrinsic nonlinear oscillatory dynamics, and that the frequency of oscillation lies within the spindle range. Spindle oscillations are an early sleep oscillation characterized by high frequency bursts of action potentials followed by a period of quiescence, at a frequency of 7-14 Hz. Spindles are generally regarded as being generated by intrathalamic circuitry, as decorticated thalamic slices and the isolated thalamic reticular nucleus exhibit spindles. However, the role of cortical feedback has been shown to regulate and synchronize the oscillation. Previous modelling studies have mainly used conductance-based models and hence the mechanism relied upon the inclusion of ionic currents, particularly the T-type calcium current. Here we demonstrate that spindle-frequency oscillatory activity can also arise from the nonlinear dynamics of the thalamocortical circuit, and we use bifurcation analysis to examine the robustness of this oscillation in terms of the functional range of the parameters used in the model. The results suggest that the thalamocortical circuit has intrinsic nonlinear population dynamics which are capable of providing robust support for oscillatory activity within the frequency range of spindle oscillations. PMID- 16367785 TI - Cardiovascular effects of noradrenaline microinjected into the dorsal periaqueductal gray area of unanaesthetized rats. AB - The periaqueductal grey area (PAG) is a mesencephalic region that is involved in the modulation of cardiovascular changes associated with behavioural responses. Among the neurotransmitters present in the PAG, noradrenaline (NA) is also known to be involved in central nervous system cardiovascular regulation. In the present study we report the cardiovascular effects of the microinjection of NA into the dorsal portion of the PAG (dPAG) of unanaesthetized rats and the peripheral mechanism involved in their mediation. Injection of NA in the dPAG of unanaesthetized rats evoked a dose-dependent pressor response accompanied by bradycardia. The magnitude of the pressor responses was higher at more rostral sites in the dPAG and decreased when NA was injected into the caudal portion of the dPAG. The responses to NA were markedly reduced in urethane-anaesthetized rats. The pressor response was potentiated by i.v. pretreatment with the ganglion blocker pentolinium and blocked by i.v. pretreatment with the vasopressin antagonist dTyr(CH2)5(Me)AVP. The results suggest that activation of noradrenergic receptors within the dPAG can evoke pressor responses, which are mediated by acute vasopressin release. PMID- 16367786 TI - Single-unit responses of serotonergic medullary raphe neurons to cardiovascular challenges in freely moving cats. AB - Single-unit activity of serotonergic neurons in the nuclei raphe obscurus (NRO) and raphe pallidus (NRP) were recorded in conjunction with heart rate in freely moving cats in response to systemic administration of vasoactive drugs and to graded haemorrhage. Bolus administration of phenylephrine hydrochloride and sodium nitroprusside (20 microg/kg, i.v.) produced a marked, transient reflex bradycardia (-42 b.p.m.) and tachycardia (+60 b.p.m.), respectively. The activity of NRO/NRP serotonergic neurons remained unchanged after phenylephrine and nitroprusside administration. The administration of hydralazine (1 mg/kg, i.v.), a long-acting vasodilator, produced sustained tachycardia (+60 b.p.m.), which was not accompanied by changes in neuronal activity, despite prolonged reflex activation of the sympathetic nervous system. The initial withdrawal of up to 15% of total blood volume increased heart rate (+12 b.p.m.), whereas the removal of 22.5% of total blood decreased heart rate (-44 b.p.m.). The activity of NRO/NRP serotonergic neurons remained unaltered throughout graded haemorrhage trials, despite the changes in sympathetic outflow. Thus, serotonergic NRO and NRP neurons appear to be insensitive to alterations in blood pressure and baroreceptor activity, and this lack of responsiveness does not support a specific role for these cells in cardiovascular regulation. Furthermore, these neurons do not appear to be involved in physiological mechanisms underlying alterations in autonomic outflow invoked by hypertension and hypotension. Taken within the context of our previous work, the present data suggest that medullary serotonergic neurons may modulate autonomic outflow, but only in relation to their primary role in motor control. PMID- 16367787 TI - Effects of systemically administered cocaine on sensory responses to peri threshold vibrissae stimulation: individual cells, ensemble activity, and animal behaviour. AB - Previous studies have shown that systemic administration of cocaine transiently alters stimulus-evoked responses of ventral posteromedial (VPM) thalamic neurons. Results from these single-unit electrophysiological studies revealed that cocaine was equally likely to augment or attenuate the magnitude of sensory evoked responses following threshold level stimulation of peripheral receptive fields. In an attempt to clarify the impact of cocaine administration on sensory signal processing, we examined the drug's effects on responses of individual neurons and ensembles of VPM thalamic neurons to sensory stimuli, and performance of subjects in a sensory detection behavioural task. Extracellular responses of single (n = 1 cell/rat) or multiple VPM thalamic neurons (n = 10-40 cells/rat) were monitored before and after cumulative doses of cocaine (0.25-2.0 mg/kg i.v.). Neuronal responses were characterized by assessing the response profile to a range of peri threshold-level deflections of the optimal whisker on the contralateral face. Drug effects on stimulus-response curves, determined from quantitative analysis of spike train data, indicated that whereas cocaine elicits variable effects at the single cell level, the stimulus-evoked response of the recorded population was likely to increase following lower (0.25-1.0 mg/kg i.v.) doses of cocaine. Furthermore, cocaine preferentially enhanced responses to smaller magnitude deflections of vibrissa, altering the response profile from a mode that accurately conveyed stimulus strength to one that increased detection at the expense of discrimination. Finally, a similar pattern emerged in a behavioural paradigm involving rats trained to detect variable amplitude whisker pad stimulation, suggesting a common action of cocaine that may contribute to the drug's addictive properties. PMID- 16367788 TI - Alterations of arcuate nucleus neuropeptidergic development in contactin deficient mice: comparison with anorexia and food-deprived mice. AB - A mutation in the Contactin-1 gene results in an ataxic and anorectic phenotype that is apparent by postnatal day 10 and lethal by postnatal day 19 [Berglund et al. (1999) Neuron 24, 739-750]. The resemblance of this phenotype with the anorexia (anx/anx) mouse mutation prompted us to investigate the hypothalamic neurochemistry of Contactin knock-out (KO) mice. Contactin was expressed in the hypothalamic neuropil of wild-type (WT) but not Contactin KO mice. In the KO condition, neuropeptide Y (NPY) and agouti-related protein (AgRP) immunoreactivity (IR) accumulated in the somata of arcuate nucleus neurons, whereas IR for these neuropeptides as well as for alpha-melanocyte-stimulating hormone (alpha-MSH) decreased in the corresponding axon projections. These changes in the pattern of neuropeptide expression in the Contactin-deficient hypothalamus were similar but more pronounced than those found in anx/anx mice. Increased levels of NPY and AgRP and decreased concentrations of pro opiomelanocortin mRNA in arcuate neurons accompanied these changes. In relating these alterations a 24-h food deprivation period, we observed in 3-week-old WT mice an elevation of NPY- and AgRP-IR in the perikarya of arcuate neurons without notable reduction of NPY- or AgRP-IR in nerve fibers, suggesting that the decrease of arcuate projections can be associated with postnatal anorectic phenotype. Our data implicate Contactin in the postnatal development of the NPY/AgRP and alpha-MSH arcuate neurons and suggest that similar to anx/anx mutant mice, compromised orexigenic signaling via NPY/AgRP neurons may contribute to reduced food intake by the Contactin-mutant animals. PMID- 16367789 TI - Striatal muscarinic receptor antagonism reduces 24-h food intake in association with decreased preproenkephalin gene expression. AB - Cholinergic interneurons of the striatum respond to motivationally relevant stimuli and are involved in appetitive learning. However, there has been relatively little inquiry into the role of striatal acetylcholine in food motivation. Here we show in rats that a single infusion of the muscarinic receptor antagonist scopolamine (0, 5.0 or 10.0 microg/0.5 microL bilaterally) potently reduced 24-h food intake following injections into either the ventral or dorsal striatum, without affecting water intake. Furthermore, muscarinic receptor blockade induced reliable and widespread reductions in striatal preproenkephalin, but not preprodynorphin, mRNA expression. These data suggest a novel role for striatal acetylcholine in modulating feeding behavior via its effects on enkephalin gene expression. As prior research indicates a critical role for striatal enkephalin in consummatory behaviors and palatability, we hypothesize that cholinergic interneurons assist in translating hypothalamic energy state signals into food-directed behaviors via their regulation of striatal opioid peptides. PMID- 16367790 TI - Down-regulation of metabotropic glutamate receptor 1alpha in globus pallidus and substantia nigra of parkinsonian monkeys. AB - Enhanced glutamatergic neurotransmission via the subthalamopallidal or subthalamonigral projection seems crucial for developing parkinsonian motor signs. In the present study, the possible changes in the expression of metabotropic glutamate receptors (mGluRs) were examined in the basal ganglia of a primate model for Parkinson's disease. When the patterns of immunohistochemical localization of mGluRs in monkeys administered systemically with 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) were analysed in comparison with normal controls, we found that expression of mGluR1alpha, but not of other subtypes, was significantly reduced in the internal and external segments of the globus pallidus and the substantia nigra pars reticulata. To elucidate the functional role of mGluR1 in the control of pallidal neuron activity, extracellular unit recordings combined with intrapallidal microinjections of mGluR1-related agents were then performed in normal and parkinsonian monkeys. In normal awake conditions, the spontaneous firing rates of neurons in the pallidal complex were increased by DHPG, a selective agonist of group I mGluRs, whereas they were decreased by AIDA, a selective antagonist of group I mGluRs, or LY367385, a selective antagonist of mGluR1. These electrophysiological data strongly indicate that the excitatory mechanism of pallidal neurons by glutamate is mediated at least partly through mGluR1. The effects of the mGluR1-related agents on neuronal firing in the internal pallidal segment became rather obscure after MPTP treatment. Our results suggest that the specific down-regulation of pallidal and nigral mGluR1alpha in the parkinsonian state may exert a compensatory action to reverse the overactivity of the subthalamic nucleus-derived glutamatergic input that is generated in the disease. PMID- 16367791 TI - Effector-independent voluntary timing: behavioural and neuroimaging evidence. AB - We investigated effector-independent aspects of voluntary motor timing, using behavioural measurements and functional magnetic resonance imaging. Two types of temporal pattern were investigated; one isochronous, the other a metric, rhythmic sequence of six temporal intervals. Each pattern was performed using tapping movements with the left or right index fingers, or rhythmic speech on one syllable. Deviations from the ideal temporal pattern in the rhythmic sequence tasks were consistent between the three different effectors, within subjects. This suggests that the same representation of the rhythm was used to time the movements with all effectors. To reveal brain regions involved in such effector independent timing, we localized the overlap in brain activity when the rhythmic sequence was performed with the different effectors. Activity was found in the mesial and lateral premotor cortices, posterior and anterior regions of the superior temporal gyrus and the inferior frontal cortex. Subcortical activations were in the left globus pallidus, the vermis and bilaterally in the cerebellar hemispheres (lobule VI) and the thalamus. The overlap in activity between the isochronous tasks included the same set of brain regions, except for the basal ganglia and the thalamus. Rhythmic sequences had significantly higher activity in mesial premotor cortex, the left superior temporal gyrus and the cerebellum, than had isochronous movements. These findings reveal a set of brain regions likely to be involved in effector-independent representations of temporal patterns in voluntary motor timing. A subset of these regions plays important roles for the organization of rhythmic sequences of several intervals. PMID- 16367792 TI - Premotor neuronal plasticity in monkeys adapting to a new dynamic environment. AB - Recent evidence indicates that premotor cortex (PM) in addition to their well established motor functions, also play a role in nonmotor processes such as spatial attention and working memory. In the present study, neuronal activities in dorsal PM (PMd) and ventral PM (PMv) were recorded in a force field adaptation task. This study found that PM neurons show learning-related plasticity and that a neuron demonstrates either one type or multiple types of properties (i.e. kinematic, dynamic, and memory). The current study reveals that memory properties could be displayed by one or a combination of the cell activity parameters [i.e. average firing rate (AFR), dynamic range (DR), and preferred direction (PD)]. A predominant percentage of cells displayed memory properties with AFR or AFR plus other parameters. This study investigated the memory properties vs. the time sequence of the task trial [i.e. delay time (DT), movement time (MT), and target holding time (THT)] and found that: (i) most neurons display memory properties only in one time window; (ii) few neurons display memory properties in three time windows, and (iii) there are significantly more cells showing memory properties during MT than during any other time windows. There are cells that show memory I (changing their tuning curves in the force field and retaining those changes after the force field was removed), memory II (changing their tuning curves after the force field was removed), or both properties. Significantly more cells display one type of memory property (memory I or memory II) rather than both types of memory properties (memory I and memory II). PMID- 16367793 TI - The role of prefrontal cortex in object-in-place learning in monkeys. AB - Previous ablation studies in monkeys suggest that prefrontal cortex is involved in a wide range of learning and memory tasks. However, monkeys with crossed unilateral lesions of frontal and temporal cortex are unimpaired at concurrent object-reward association learning but are impaired at conditional learning and the implementation of memory-based performance rules. We trained seven monkeys preoperatively on an associative learning task that required them to associate objects embedded in unique complex scenes with reward. Three monkeys then had crossed unilateral lesions of frontal and inferior temporal cortex and the remaining monkeys had bilateral prefrontal cortex ablation. Both groups were severely impaired postoperatively. These results show that both bilateral prefrontal cortex ablation and frontal-temporal disconnection impair associative learning for objects embedded in scenes. The results provide evidence that the function of frontal-temporal interactions in memory is not limited to conditional learning tasks and memory-dependent performance rules. We propose that rapid object-in-place learning requires the interaction of frontal cortex with inferotemporal cortex because visual object and contextual information which is captured over multiple saccades must be processed as a unique complex event that is extended in time. The present results suggest a role for frontal-temporal interaction in the integration of visual information over time. PMID- 16367795 TI - Neural stem and progenitor cells: biology and clinical potential, part two. Proceedings of a symposium. July 2004. Cork, Ireland. PMID- 16367794 TI - Compartmentalization of calcium entry pathways in mouse rods. AB - Photoreceptor metabolism, gene expression and synaptic transmission take place in a highly polarized structure consisting of the ellipsoid, subellipsoid, cell body and synaptic terminal regions. Although calcium, a key second messenger, regulates cellular functions throughout the photoreceptor, the molecular mechanisms underlying local region-specific action of Ca2+ in photoreceptors are poorly understood. I have investigated the compartmentalization of voltage dependent and independent Ca2+ channels in mouse photoreceptors. Transient receptor potential channels isoform 6 (TRPC6), a putative store-operated Ca2+ channel, was selectively localized to the cell body of rods. By contrast, voltage operated Ca2+ channels were expressed in the synaptic terminal and in the ellipsoid/subellipsoid regions. Likewise, Ca2+ store transporters and channels were strongly associated with the subellipsoid region. A moderate TRPC6 signal was observed in cell bodies of bipolar, amacrine and ganglion cells, but was absent from both plexiform layers. These results suggest that Ca2+ entry mechanisms in mammalian photoreceptors and bipolar cells are highly compartmentalized, consistent with local, region-specific activation of Ca2+ dependent processes. PMID- 16367796 TI - Astrocytes and NG2-glia: what's in a name? AB - Classic studies recognize two functionally segregated macroglial cell types in the central nervous system (CNS), namely astrocytes and oligodendrocytes. A third macroglial cell type has now been identified by its specific expression of the NG2 chondroitin sulphate proteoglycan (NG2-glia). These NG2-glia exist abundantly in both grey and white matter of the mature CNS and are almost as numerous as astrocytes. It is well established that NG2-glia give rise to oligodendrocytes. However, the majority of NG2-glia in the adult CNS proliferate very slowly and are non-motile. Both astrocytes and NG2-glia display a stellate morphology and express ion channels and receptors to neurotransmitters used by neurons. Both types of glia make intimate contacts with neurons in grey and white matter, and their functional differences and similarities are only beginning to be unravelled. Recent observations emphasize the need to examine the relationship between astrocytes and NG2-glia, and address the question of whether they represent overlapping or two distinct glial cell populations. To be of any relevance, this classification must relate to specific functions in the neural network. At present, the balance of evidence is that NG2-glia and astrocytes are functionally segregated populations. PMID- 16367797 TI - Synantocytes: the fifth element. AB - Classic studies have recognized neurons and three glial elements in the central nervous system (CNS) - astrocytes, oligodendrocytes and microglia. The identification of novel glia that specifically express the NG2 chondroitin sulphate proteoglycan (CSPG) raises the possibility of a fifth element. Until recently, all NG2-expressing glia were considered to be oligodendrocyte precursor cells (OPCs) that persist in the adult CNS to generate oligodendrocytes throughout life. However, this narrow view of the function of 'NG2-glia' is being challenged. The majority of NG2-expressing glia in the adult CNS are a distinct class of cells that we have called 'synantocytes' (from the Greek synanto for contact). Synantocytes are stellate cells, with large process arborizations, and are exquisitely related to neurons. Individual cells traverse white and grey matter and form multiple contacts with neurons, astrocytes, oligodendrocytes and myelin. Synantocytes are an integral component of the 'tetrapartite' synapse, and provide a potential integrative neuron-glial communications pathway. Neuronal activity, glutamate and adenosine triphosphate (ATP) act on synantocyte receptors and evoke raised intracellular calcium. It remains to be seen whether this serves a physiological function, but synantocytes may be specialized to monitor signals from neurons and glia, and to respond to changes in the integrity of the CNS via their specific contacts and ion channel and receptor profiles. The general consequences of synantocyte activation are proliferation and phenotypic changes, resulting in glial scar formation, or regeneration of oligodendrocytes, and possibly neurons. PMID- 16367798 TI - NG2-expressing cells as oligodendrocyte progenitors in the normal and demyelinated adult central nervous system. AB - The mammalian adult central nervous system (CNS) is known to respond rapidly to demyelinating insults by regenerating oligodendrocytes for remyelination from a dividing precursor population. A widespread population of cells exists within the adult CNS that is thought to belong to the oligodendrocyte lineage, but which do not express proteins characteristic of mature myelinating oligodendrocytes, such as myelin basic protein (MBP) and 2,3-cyclic nucleotide 3-phosphodiesterase (CNP). Instead, these cells have phenotypic characteristics of a more immature stage of the oligodendrocyte lineage. They express the NG2 chondroitin sulphate proteoglycan, in addition to O4 and the platelet-derived growth factor alpha receptor, all widely accepted as markers for oligodendrocyte progenitor cells (OPCs) throughout development. However, NG2+ cells residing in the adult CNS do not resemble embryonic or neonatal NG2+ cells in terms of their morphology or proliferation characteristics, but instead represent a unique type of glial cell that has the ability to react rapidly to CNS damage. In this review, we present the evidence that adult NG2+ cells are part of the oligodendrocyte lineage and are capable of giving rise to new oligodendrocytes under both normal and demyelinating conditions. We also review the literature that these cells may have multiple functional roles within the adult CNS, notwithstanding their primary role as OPCs. PMID- 16367799 TI - NG2: a component of the glial scar that inhibits axon growth. AB - NG2 is a high-molecular-weight chondroitin sulphate proteoglycan found on the surfaces of oligodendrocyte precursor cells (OPCs). Here we review the history and biology of OPCs with an emphasis on their functions after experimentally induced CNS injury. Injury to brain or spinal cord results in the rapid accumulation of NG2-expressing OPCs in the glial scar that forms at the injury site. The glial scar is considered a biochemical and physical barrier to successful axon regeneration and the functional properties of NG2 suggest that it, along with other macromolecules, participates in the creation of this growth inhibitory environment. NG2 is an important target for therapies designed to promote successful axon regrowth. PMID- 16367800 TI - Instructive niches: environmental instructions that confound NG2 proteoglycan expression and the fate-restriction of CNS progenitors. AB - Cellullar deficits are replenished within the central nervous system (CNS) by progenitors to maintain integrity and recover function after injury. NG2 proteoglycan-expressing progenitors replenish oligodendrocyte populations, but the nature of NG2 proteoglycan may not indicate a restricted population of progenitors. After injury, restorative spatiotemporal cues have the potential ability to regulate divergent fate-choices for NG2 progenitors, and NG2 progenitors are known to produce multiple cell types in vitro. Recent data suggest that NG2 expression is attenuated while protein levels remain high within injurious tissue; thus, NG2 expression is not static but transiently controlled in response to a dynamic interplay of environmental cues. Therefore, NG2 proteoglycan expression could label newly generated cells or be inherited by resident cell populations that produce oligodendrocytes for remyelination, astrocytes that provide trophic support and other cells that contribute to CNS function. PMID- 16367801 TI - NG2-expressing cells in the nervous system: role of the proteoglycan in migration and glial-neuron interaction. AB - The NG2 glycoprotein is a type I membrane protein expressed in the developing and adult central nervous system (CNS) by subpopulations of glia including oligodendroglial precursor cells (OPCs), and in the developing CNS additionally by pericytes. In the mouse CNS, expression of NG2 protein is already observed at embryonic day 13 and peaks between postnatal days 8 and 12. NG2+ cells persist in grey and white matter in adult mouse brain: cells in the developing and adult brain show clear differences in migration, cell-cycle length and lineage restriction. Several groups have provided evidence that subpopulations of NG2+ cells can generate neurons in vivo. Neuronal stimulation in the developing and adult hippocampus leads to Ca2+ signals in apposing NG2+ glia, suggesting that these cells may modulate synaptic activity, and NG2+ cells often ensheath synapses. The structure of the protein with two N-terminal LamininG/Neurexin/Sex hormone-binding globulin domains suggests a role in adhesion. The C-terminal PSD 95/DiscsLarge/Zona Occludens-1 (PDZ)-binding motif has been found to associate with several PDZ proteins including the Glutamate Receptor Interacting Protein GRIP: NG2 may thus act to position AMPA receptors on glia towards sites of neuronal glutamate release. Furthermore, the NG2 proteoglycan plays a role in cell migration and spreading and associates with actin-containing cytoskeletal structures. PMID- 16367802 TI - Characterization of the rhesus monkey superior olivary complex by calcium binding proteins and synaptophysin. AB - This study was performed in order to characterize the main nuclei of the rhesus monkey superior olivary complex by means of antibodies against the calcium binding proteins parvalbumin, calbindin and calretinin and the synaptic vesicle protein synaptophysin. These markers revealed the neuronal morphology and organization of nuclei located within the rhesus monkey superior olivary complex. The architectural details included the distribution of axonal terminals on neurons. The medial superior olivary nucleus was present as a column of neurons. No clear segregation of calretinin-positive terminals was noticed on the medial and lateral dendritic fields of these neurons. The lateral superior olivary nucleus was characterized by a distinct nuclear shape. Calretinin-, parvalbumin- or calbindin-positive terminals contacted somata and dendrites. The medial nucleus of trapezoid body could be clearly differentiated as a distinct region in the rhesus monkey superior olivary complex. Somata of that nucleus showed calbindin- and parvalbumin-labelling whereas somatic calyces of Held were reavealed by calretinin and synaptophysin labelling. The results are discussed with respect to the processing of acoustic information in primate species and their ability to hear high and low frequencies, which is reflected by anatomical correlates. PMID- 16367803 TI - Fetal programming of fat and collagen in porcine skeletal muscles. AB - Connective tissue plays a key role in the scaffolding and development of skeletal muscle. Pilot studies carried out in our laboratory have shown that the smallest porcine littermate has a higher content of connective tissue within skeletal muscle compared with its largest littermate. The present study investigated the prenatal development of intralitter variation in terms of collagen content within connective tissue and intramuscular fat of the M. semitendinosus. Twenty-three pairs of porcine fetuses from a Large White-Landrace origin were used aged from 36 to 86 days of gestation. The largest and smallest littermates were chosen by weight and the M. semitendinosus was removed from each. Complete transverse muscle sections were stained with Oil Red O (detection of lipids) and immunocytochemistry was performed using an antibody to collagen I. Slides were analysed and paired t-Tests revealed the smallest littermate contained a significantly higher proportion of fat deposits and collagen I content compared with the largest littermate. Recent postnatal studies showing elevated levels of intramuscular lipids and low scores for meat tenderness in the smallest littermate corroborate our investigations. It can be concluded that the differences seen in connective tissue elements have a fetal origin that may continue postnatally. PMID- 16367804 TI - Changes in adipocytes and dendritic cells in lymph node containing adipose depots during and after many weeks of mild inflammation. AB - The time course and cellular basis for inflammation-induced hypertrophy of adipose tissue were investigated over 20 weeks in mature male rats. Mild inflammation was induced by subcutaneous injection of 20 microg lipopolysaccharide into one hind-leg three times/week for 4 or 8 weeks, followed by up to 12 weeks 'rest' without intervention. Mean volume and frequency of apoptosis (TUNEL assay) were measured in adipocytes isolated from sites defined by their anatomical relations to lymph nodes, plus numbers of CCL21-stimulated lymph node-derived and adipose tissue-derived dendritic cells. Experimental inflammation increased dendritic cells and adipocyte apoptosis in the locally stimulated popliteal depot and the lymphoid tissue-associated regions of the contralateral popliteal and mesentery and omentum. Responses declined slowly after inflammation ended, but all measurements from the locally stimulated popliteal depot, and the omentum, were still significantly different from controls after 12 weeks rest. The locally stimulated popliteal adipose tissue enlarged by 5% within 4 weeks and remained larger than the control. We conclude that prolonged inflammation induces permanent enlargement, greater adipocyte turnover and increased dendritic cell surveillance in the adjacent adipose tissue and the omentum. The experiment suggests a mechanism for selective hypertrophy of lymphoid tissue-associated adipose tissue in chronic stress and inflammatory disorders, including impaired lymph drainage, Crohn's disease and HIV-associated lipodystrophy, and a link between evolutionary fitness, sexual selection and aesthetically pleasing body symmetry. It would be useful for further study of molecular mechanisms in inflammation-induced local hypertrophy of adipose tissue and development of specific therapies that avoid interference with whole-body lipid metabolism. PMID- 16367805 TI - Ultrastructural changes in the interhaemal membrane and junctional zone of the murine chorioallantoic placenta across gestation. AB - The mouse is an extremely useful experimental model for the study of human disease owing to the ease of genetic and physiological manipulation. A more detailed knowledge of murine placental development will, we hope, increase our understanding of the pathogenesis of placentally related complications of human pregnancy. The murine placenta consists of two main fetally derived compartments: the labyrinthine zone and the junctional zone. Exchange in the labyrinthine zone takes place across an interhaemal membrane comprising an outer layer of cytotrophoblast cells and two inner layers of syncytial trophoblast. The cytotrophoblast layer thins as gestation advances, and in addition becomes highly perforated after embryonic day (E)12.5. Furthermore, as gestation advances cytotrophoblast nuclear volume and DNA content increase, suggesting the formation of labyrinthine trophoblast giant cells. The syncytial layers become increasingly microvillous, enlarging the surface area for exchange. Separate basement membranes support the syncytium and the fetal capillary endothelium throughout gestation, although these appear to fuse where the capillaries are closely approximated to the trophoblast. The junctional zone consists of two principal trophoblast cell types, spongiotrophoblasts and invasive glycogen cells, yet the functions of each remain elusive. Spongiotrophoblasts vary in their appearance even when not fully differentiated, but a striking feature is the extensive endoplasmic reticulum of the more mature cells. Early glycogen cells are distinguished by the presence of electron-dense glycogen granules, and large amounts of surrounding extracellular matrix. Later the accumulations of glycogen granules occupy almost all the cytoplasm and there are few organelles. This is the first study to use both scanning and transmission electron microscopy in an ultrastructural description of murine placental development and is complementary to contemporary genetic investigations. PMID- 16367806 TI - Atypical development of Sertoli cells and impairment of spermatogenesis in the hypogonadal (hpg) mouse. AB - Testes of hypogonadal (hpg) mice show arrested postnatal development due to congenital deficiencies of gonadotrophin-releasing hormone (GnRH) and gonadotrophin synthesis and secretion. Follicle-stimulating hormone (FSH), androgen or oestrogen treatment restore qualitatively normal spermatogenesis in hpg testes. Understanding the cellular and molecular changes accompanying hormone induced spermatogenesis in hpg mice requires detailed morphological analyses of the germ cells and Sertoli cells in the untreated hpg testis. We compared seminiferous epithelial cytology in adult hpg, immature and adult wild-type mice using unbiased optical disector-based stereology, immunolocalization of Sertoli cell microtubules (MT), espin (a component of the blood-testis barrier), markers of Sertoli cell maturity (p27(kip1) and WT-1), and electron microscopy. Hpg testes had marked reductions in weight, seminiferous cord volume and length, and severe spermatogenic impairment with germ cells per testis < 1% of adult wild type testes. Sertoli cell nuclei expressed WT-1 in hpg testes, but often were centrally located, similar to 9-14-day-old wild-type testes, and they expressed p27(kip1), indicating that hpg Sertoli cells were post-mitotic. Hpg testes had significantly (P < 0.05) reduced Sertoli cells per testis (0.56 million) compared with 10-day wild-type (1.15 million) and adult wild-type testes (2.06 million). Immunofluorescence labelling of normal adult Sertoli cells showed supranuclear MT columns and basally located espin, but these features were absent in 10-day-old and hpg Sertoli cells. Hpg Sertoli cells showed pleomorphic nuclear ultrastructure with mature-type nucleoli, similar to normal adult-type Sertoli cells, but hpg Sertoli cells exhibited incomplete tight junctions that lacked ectoplasmic specializations. We conclude that in hpg mice, chronic gonadotrophin insufficiency restrains Sertoli cell proliferation and maturation, forming pseudo adult-type Sertoli cells that are incapable of supporting germ cell proliferation and maturation. PMID- 16367807 TI - A stereological study of the renal glomerular vasculature in the db/db mouse model of diabetic nephropathy. AB - In diabetic nephropathy, glomerular hypertrophy is evident early in response to hyperglycaemia. Alterations of capillary length and vascular remodelling that may contribute to glomerular hypertrophy and the subsequent development of glomerulosclerosis remain unclear. The present study used the db/db mouse model of Type 2 diabetes to examine the glomerular microvascular changes apparent with long-term diabetic complications. Unbiased stereological methods and high resolution light microscopy were used to estimate glomerular volume, and glomerular capillary dimensions including length and surface area in 7-month-old db/db diabetic mice and age-matched db/m control mice. The db/db diabetic mice showed significant glomerular hypertrophy, corresponding with elevated blood glucose levels, and increased body weight and kidney weight, compared with db/m control mice. Glomerular enlargement in db/db mice was associated with increases in the surface area (5.387 +/- 0.466 x 10(4) microm2 vs. 2.610 +/- 0.287 x 10(4) microm2; P < 0.0005) and length (0.3343 +/- 0.022 x 10(4) microm vs. 0.1549 +/- 0.017 x 10(4) microm; P < 0.0001) of capillaries per glomerulus, compared with non-diabetic mice. Stereological assessment at the electron microscopic level revealed increased glomerular volume density of mesangial cells and mesangial matrix, and thickening of the glomerular basement membrane in db/db mice. These results demonstrate that glomerular hypertrophy evident in advanced diabetic nephropathy in this model is associated with increased length and surface area of glomerular capillaries. The contribution of angiogenesis and vasculogenesis to the glomerular microvascular alterations in response to hyperglycaemia remain to be determined. PMID- 16367810 TI - Effects of repeated low-dose UVB irradiation on the hyphal growth of Candida albicans. AB - Ultraviolet B light (UVB) can have negative phototropic effects on fungi. Candida albicans is often found on human skin exposed to UVB. Therefore, it is of medical interest to know whether a negative phototropic response to UVB irradiation can support an invasive growth of this potentially dangerous agent. In our study we investigated how repeated irradiation with low doses of UVB can influence the hyphal growth of C. albicans. Six randomly chosen strains of C. albicans were tested. Formation of hyphae was induced and maintained within transparent agar plates. The fungi were exposed to UVB three times daily for 7 days from either the observe or the reverse side during incubation. The wavelength spectrum was in the range of 310-315 nm, single doses were between 0.0018 and 0.432 J cm(-2). After 7 days the morphology and growth direction of C. albicans cells were determined microscopically. All six strains showed a common and dose-dependent response to UVB irradiation: the progression of hyphal growth was inhibited, no phototropic effects were seen and as a new finding an increased formation of blastospores was observed. We conclude that an irradiation of human skin colonized by C. albicans with doses of UVB that can occur under natural or artificial conditions is unlikely to trigger skin invasion by C. albicans. PMID- 16367811 TI - Incubation of human chorioamniotic membranes with Candida albicans induces differential synthesis and secretion of interleukin-1beta, interleukin-6, prostaglandin E, and 92 kDa type IV collagenase. AB - Ascendant colonization of pathogenic microorganisms from the vagina to the uterus is strongly associated to preterm labour and premature rupture of membranes. This study evaluated the secretion of interleukin (IL)-1beta, tumour necrosis factor (TNF)alpha, IL-6, prostaglandin E(2) (PGE(2)), and metalloproteinases 9 and 2 by the human chorioamnion stimulated with Candida albicans. Chorioamniotic membranes were obtained after delivery by elective Cesarean section from women at 37-40 weeks of gestation without evidence of active labour. The membranes were mounted in Transwell devices that form two independent compartments, which allow testing the individual responses and contributions of the amnion and choriodecidua. One million CFU ml(-1) of C. albicans was added to either the amniotic or choriodecidual surface and secretions of the markers were measured in both compartments using specific enzyme-linked immunosorbent assay and zymography. Fetal membranes followed different secretion patterns of proinflammatory cytokines depending on the side to which the stimulus was applied. IL-1beta was produced in higher amounts in the presence of C. albicans when applied to the choriodecidual side; TNFalpha and IL-6 secretion did not change in either the amnion or choriodecidual region. PGE(2) synthesis depicted a different pattern, the amniotic tissue was more responsive than the choriodecidual tissue, and this response tended to be higher even when only the amniotic side was stimulated. Matrix metalloproteinases (MMP)-9 increased after stimulation, being the choriodecidua its main source. Selective stimulation with C. albicans induced a differential secretion of IL-1beta, PGE(2), and MMP-9, resulting from a cooperative and bidirectional communication between the amnion and the choriodecidua. PMID- 16367812 TI - An in vitro evaluation of the adhesion of Candida species to oral and lung tissue cells. AB - The analysis of the adherence capacity of fungi to surfaces of both oral tissue and different tissues would be of interest in the fungal dissemination as an oral and systemic pathogen. We developed an in vitro adenosine triphosphate (ATP) based assay technique to extract the cellular and fungal ATP separately, which allowed the quantitative evaluation of the adhesion of the yeast to monolayers of human gingival epithelial cells (GEC), gingival fibroblasts (GF) and pulmonary fibroblasts (PF). Seven oral isolates of Candida species (three of Candida albicans, three of Candida tropicalis and one of Candida glabrata) were used in the study. The adherent level of the Candida species varied depending on both the isolates and the cell origins, although all the Candida isolates had a significantly higher level of adherence to GEC than to GF except the single isolate of C. tropicalis. Whereas the adherent level of the five isolates to GEC was significantly higher than that to PF, the adherent level of the remaining two isolates of C. tropicalis to GEC was significantly lower than that to PF. These results suggest that candidal adherence to host tissue cells should be regulated in an isolate-dependent and cell-origin-dependent manner, and that the phenomena may be involved in the colonisation and/or dissemination of the fungi. PMID- 16367813 TI - Comparison of polar lipids from yeast and mycelial forms of Candida albicans and Candida dubliniensis. AB - The aim of the present study was to compare polar lipids of yeast and mycelial forms of both Candida albicans and Candida dubliniensis. Cultures were harvested from Lee's medium after incubation at 37 degrees C for 48 h. Yeast and mycelial forms were washed, separated from one another, dried and lipids extracted and prepared for fast atom bombardment mass spectrometry analysis in the negative-ion mode. For fatty acids, differences between the yeast and mycelial forms were greater for C. dubliniensis than for C. albicans. For the phospholipid families, phosphatidic acid and phosphatidylethanolamine, differences between yeast and mycelial forms were greater for C. dubliniensis than for C. albicans. For both species, it is concluded that both fatty acid and phospholipid molecular species compositions differ according to whether the cells are in the yeast or mycelial form. PMID- 16367814 TI - Cockroaches as carriers of fungi of medical importance. AB - The fungal pathogenic flora of the external surface of 103 cockroaches (Periplaneta americana) collected from the intensive care unit of a hospital were investigated. In this study, a high percentage of test cockroaches (93.2%) were found to carry fungi of medical importance. The main fungi isolated were species of Candida, Aspergillus and Penicillium. Information about the carriage of pathogenic fungi by cockroaches in hospital environment is scanty. The results suggest that cockroaches can play a role in dissemination of fungi, which they can carry on their external surface. PMID- 16367815 TI - Comparison of diagnostic methods in the diagnosis of dermatomycoses and onychomycoses. AB - Direct microscopic examination of potassium hydroxide (KOH)-prepared specimens is the simplest, cheapest method used for the diagnosis of mycotic infections of the skin. However, KOH preparations have been reported to have 5-15% of false negative results, possibly because of the low visibility of scant, scattered fungal material of the nail scrapings and because the detection of fungal elements depends on the skill of the observer [Arch Dermatol133 (1997) 1317; Clin Microbiol Rev8 (1995) 240]. We compared two different KOH-based staining methods in order to obtain reliable results in shorter time than expected for cultures. A total of 124 patients with suspect diagnosis of dermatomycosis or onychomycosis were enrolled. Two scrapings from the same lesion of each patient were stained with KOH-Chlorazole and KOH-Acridine Orange (AO), respectively; cultural examination of the same specimen was considered as diagnostic gold standard. The two methods showed neither significantly different sensitivity nor specificity; however, for onychomycoses, we observed a slightly higher sensitivity for KOH Chlorazole and a higher specificity for KOH-AO. We suggest the use of both techniques in order to improve detection of fungal infection, especially for onychomycoses. PMID- 16367816 TI - Invasive fungal sinusitis of isolated sphenoid sinus in immunocompetent subjects. AB - Invasive fungal sinusitis of the paranasal sinuses in a healthy immunocompetent person is uncommon. Isolated involvement of any paranasal sinus, particularly sphenoid sinus is rare. In this study, five immunocompetent patients who had no nasal complaints but obscure symptoms of headache and orbital symptoms such as diplopia, retro-orbital pain and loss of vision were diagnosed to be having fulminant fungal sinusitis of the sphenoid sinus. Three patients had aspergillosis and two patients had mucormycosis. These patients initially presented to neurologists and ophthalmologists because they had no ENT complaints. The diagnosis was made on endoscopy, radiology and histopathology. They were treated aggressively according to the standard protocols. The purpose of this paper is to bring to light the changing clinical spectrum of invasive fungal sinusitis. It can occur in immunocompetent patients and in the form of isolated sphenoid sinus involvement. PMID- 16367817 TI - Pseudomembranous and obstructive Aspergillus tracheobronchitis - optimal diagnostic strategy and outcome. AB - Pseudomembranous and obstructive Aspergillus tracheobronchitis (PMATB/OATB) are still considered to be refractory to therapy and to have a fatal outcome. To evaluate the optimal diagnostic strategy and to describe factors affecting the outcome of PMATB and OATB. Retrospective analysis of four new cases of PMATB and OATB combined with 16 previously reported cases over a 10-year period (1995 2004). Among the four new cases reported and the 16 published cases, four patients survived their infection. The mortality rate was significantly higher in the group of ventilated patients [94% (15 of 16 patients)] than in the group of non-ventilated patients [25% (1 of 4 patients), P < 0.05, Fisher's exact test]. In all 20 patients, diagnosis was established by bronchoscopy. Culture examination of mucous plugs was positive in 8 of 10, culture of the tracheobronchial aspirate was positive in 8 of 12, and bronchoalveolar lavage was diagnostic in 7 of 13 patients. All bronchoscopic techniques were complementary in improving the yield of bronchoscopy. However, microscopy of mucous plugs and/or necrotic material was the best diagnostic modality [positive in 94% (17 of 18 patients)]. Prognosis of PMATB and OATB remains poor. Microscopy of respiratory specimens is the most sensitive tool to confirm the diagnosis. The characteristic appearance of the disease makes it possible to start antifungal therapy immediately. PMID- 16367818 TI - Candida endocarditis in neonates: report of five cases and review of the literature. AB - Candidal endocarditis is an uncommon and serious complication of invasive Candida infection in neonates. The aim of this study was to further characterise candidal endocarditis in neonates. Between 1995 and 2000, 56 patients were diagnosed with Candida bloodstream infections (CBSI) in the Neonatal Intensive Care Unit of Schneider Children's Medical Center of Israel. Five of them (9%) developed mycetoma of the right atrium. None of the patients had congenital heart disease or a central venous catheter in the right heart at the time of diagnosis. All were treated with amphotericin B alone or in combination with other antifungals, without surgical intervention. One patient died of the disease and one died later of polymicrobial sepsis and necrotizing enterocolitis. A review of the literature since 1980 yielded an additional 25 cases of candidal endocarditis. For the whole sample (n = 30) survival rate was 73.1%. Six of the 10 patients treated with antifungal agents and surgery survived (60%), compared with 13 of the 20 patients treated only medically (65%) (P = 1.0). Candida endocarditis in neonates differs from fungal endocarditis in adults in risk factors, clinical presentation and outcome. As the outcome of surgical and medical treatment are comparable, antifungal therapy alone may be a valid therapeutic option in high-risk cases. PMID- 16367819 TI - Etiology of fungaemia and catheter colonisation in Argentinean paediatric patients. AB - Yeast strains obtained from blood cultures and catheters from intensive care units (ICU) and hospitalised oncology paediatrics were studied. Yeast were the first cause of catheter colonisation (51/627), and the third cause of bloodstream infection (44/6065). In catheter, the most frequent species were Candida albicans (34%), C. parapsilosis (27.7%) and C. tropicalis (15%). In blood, C. albicans (40.8%), C. parapsilosis (26.6%), C. tropicalis (15%). Malassezia furfur and Malassezia sympodialis were isolated from catheters from ICU patients. All isolates were susceptible to amphotericin B, 88.8% to itraconazole and 91.9% to fluconazole. Candida albicans and C. tropicalis strains resistant to fluconazole and itraconazol were detected. These results reveal a change in the predominant role of C. albicans as cause of candidemia in hospitalised children and the emergence of antifungal resistant species. These variations emphasise the importance of performing a permanent surveillance to observe and assess them. PMID- 16367820 TI - Onychomycosis is no longer a rare finding in children. AB - Onychomycosis was considered uncommon in children. This survey was carried out to estimate the frequency of fungal nail infections in children and adolescents (0 18 years of age) attending our clinic in the last decade and gain more insight into the aetiology and clinics of this entity in the paediatric age group. This study is based on data obtained from 2320 children and adolescents suspected of superficial fungal infection. Onychomycosis was diagnosed in 99 cases, representing 19.8% of all mycologically confirmed superficial mycoses (500 cases) in our material. Fingernail onychomycosis was recognized in 52 (10.4%) cases; children under 3 years of age were predominantly involved. Candida albicans was the most common isolated pathogen. Toenail onychomycosis concerned 47 (9,4%) patients; the incidence increased steadily with increasing age. Trichophyton rubrum was the most common aetiological agent with respect to toenail infection followed by T. mentagrophytesvar. interdigitale and T. mentagrophytes var. granulosum. The majority of fungal nail infections were characterized clinically by distal and lateral subungual onychomycosis. The growing trend towards the frequency of toenail and fingernail onychomycosis in children and adolescents was found in the last decade in north Poland. The results of our study show that onychomycosis in prepubertal children is not exceptional and should be considered in differential diagnosis of nail plate disorders. PMID- 16367821 TI - Initial Candida dubliniensis isolate in Candida spp. positive haemocultures in Turkey between 2001 and 2004. AB - Candida dubliniensis which was first recognized in 1995 can be easily misidentified because of its phenotypic similarities with Candida albicans. In this study blood samples of patients from various departments of Ankara University Medical Faculty between January 2001-June 2004 were investigated for the distribution of Candida spp. and the presence of C. dubliniensis. Culture positive 67 fungi were included to the study. Phenotypic tests such as chlamydospore formation, colony morphology on Staib agar, growth at 45 degrees C, carbohydrate assimilation profiles were investigated for identification and differentiation of C. dubliniensis from C. albicans. To confirm the results polymerase chain reaction were used for suspected C. albicans and C. dubliniensis isolates. Among 38 germ tube and chlamydospore forming isolates, 37 of them were found as C. albicans and one as C. dubliniensis. The incidence of C. dubliniensis in our hospital is still low, this is the first C. dubliniensis isolate as an agent of candidaemia reported from Turkey. PMID- 16367822 TI - Aetiological agents of tinea capitis in Tehran (Iran). AB - A total of 1568 patients with suspected tinea capitis were examined for causative fungal agents between 1994 and 2001. Laboratory examination confirmed tinea capitis in 209 patients. Males were affected more frequently (67.5%) than females (32.5%) and in both sexes, those who were 3-11 years old, were more infected. Trichophyton violaceum was the most common aetiological agent (37.3%) followed by Trichophyton schoenleinii (21.5%), Microsporum canis (18.6%), Trichophyton verrocosum (14.8%), Trichophyton tonsurans (5.3%), Trichophyton rubrum (1%), Microsporum gypseum (1%) and Trichophyton mentagrophytes (0.5%). A higher incidence of the disease was found to be correlated with larger family and class size. The findings are discussed in relation to different socioeconomic and hygienic backgrounds of the children. PMID- 16367823 TI - Smoking and cryptococcosis in AIDS patients. PMID- 16367826 TI - Rapid parallel evolutionary changes of gene transcription profiles in farmed Atlantic salmon. AB - Farmed salmon strains have been selected to improve growth rates as well as other traits of commercial interest but the 2 million farmed salmon escaping annually may enhance the risk of extinction of wild populations through genetic and ecological interactions. Here, we compare the transcription profiles of 3557 genes in the progeny of farmed and wild Atlantic salmon from Norway and Canada grown in controlled conditions, and demonstrate that five to seven generations of artificial selection led to heritable changes in gene transcription profiles, the average magnitude of the differences being 25% and 18% for at least 1.4% and 1.7% of the expressed genes in juvenile salmon from Norway and Canada, respectively. Moreover, genes showing significant transcription profile differences in both farmed strains (16%) all exhibited parallel changes. These findings, along with the identification of several genes whose expression profiles were modified through artificial selection, provide new insights into the molecular basis of parallel evolution, and suggest how gene flow from farmed escapees may affect the genetic integrity of wild populations. PMID- 16367827 TI - Life history correlates of inbreeding depression in mandrills (Mandrillus sphinx). AB - Inbreeding depression reflects the negative consequences of increased homozygosity at genes that affect fitness. We investigate inbreeding depression in a semi-free-ranging colony of mandrills (Mandrillus sphinx), using high quality pedigree data, comprising five maternal generations and 20 years of morphological and demographic data. We examine the relationship between inbreeding coefficients and four fitness correlates: two growth parameters (mass and height for age) and longevity in both sexes, and age at first conception in females. Inbreeding was correlated with both growth parameters, but only in females, with inbred females being smaller than noninbred females. Inbreeding was also correlated significantly with age at first conception, with inbred females giving birth earlier in life than noninbred females. We suggest that sex-biased maternal investment may explain this sex-differential response to inbreeding, although the lack of a significant association between inbreeding and growth in males may also be due to the provisioned nature of the colony. The surprising relationship between age at first conception and inbreeding may be related to smaller adult size in inbred females, or to their being less able to escape from male sexual coercion. PMID- 16367828 TI - Comparative population structure of Cynopterus fruit bats in peninsular Malaysia and southern Thailand. AB - The extent to which response to environmental change is mediated by species specific ecology is an important aspect of the population histories of tropical taxa. During the Pleistocene glacial cycles and associated sea level fluctuations, the Sunda region in Southeast Asia experienced concurrent changes in landmass area and the ratio of forest to open habitat, providing an ideal setting to test the expectation that habitat associations played an important role in determining species' response to the opportunity for geographic expansion. We used mitochondrial control region sequences and six microsatellite loci to compare the phylogeographic structure and demographic histories of four broadly sympatric species of Old World fruit bats in the genus, Cynopterus. Two forest-associated species and two open-habitat generalists were sampled along a latitudinal transect in Singapore, peninsular Malaysia, and southern Thailand. Contrary to expectations based on habitat associations, the geographic scale of population structure was not concordant across ecologically similar species. We found evidence for long and relatively stable demographic history in one forest and one open-habitat species, and inferred non-coincident demographic expansions in the second forest and open-habitat species. Thus, while these results indicate that Pleistocene climate change did not have a single effect on population structure across species, a correlation between habitat association and response to environmental change was supported in only two of four species. We conclude that interactions between multiple factors, including historical and contemporary environmental change, species-specific ecology and interspecific interactions, have shaped the recent evolutionary histories of Cynopterus fruit bats in Southeast Asia. PMID- 16367829 TI - The relative influence of natural selection and geography on gene flow in guppies. AB - Two general processes may influence gene flow among populations. One involves divergent selection, wherein the maladaptation of immigrants and hybrids impedes gene flow between ecological environments (i.e. ecological speciation). The other involves geographic features that limit dispersal. We determined the relative influence of these two processes in natural populations of Trinidadian guppies (Poecilia reticulata). If selection is important, gene flow should be reduced between different selective environments. If geography is important, gene flow should be impeded by geographic distance and physical barriers. We examined how genetic divergence, long-term gene flow, and contemporary dispersal within a watershed were influenced by waterfalls, geographic distance, predation, and habitat features. We found that waterfalls and geographic distance increased genetic divergence and reduced dispersal and long-term gene flow. Differences in predation or habitat features did not influence genetic divergence or gene flow. In contrast, differences in predation did appear to reduce contemporary dispersal. We suggest that the standard predictions of ecological speciation may be heavily nuanced by the mating behaviour and life history strategies of guppies. PMID- 16367830 TI - Efficiency of model-based Bayesian methods for detecting hybrid individuals under different hybridization scenarios and with different numbers of loci. AB - Accurate detection of offspring resulting from hybridization between individuals of distinct populations has a range of applications in conservation and population genetics. We assessed the hybrid identification efficiency of two methods (implemented in the STRUCTURE and NEWHYBRIDS programs) which are tailored to identifying hybrid individuals but use different approaches. Simulated first- and second-generation hybrids were used to assess the performance of these two methods in detecting recent hybridization under scenarios with different levels of genetic divergence and varying numbers of loci. Despite the different approaches of the methods, the hybrid detection efficiency was generally similar and neither of the two methods outperformed the other in all scenarios assessed. Interestingly, hybrid detection efficiency was only minimally affected by whether reference population allele frequency information was included or not. In terms of genotyping effort, efficient detection of F1 hybrid individuals requires the use of 12 or 24 loci with pairwise F(ST) between hybridizing parental populations of 0.21 or 0.12, respectively. While achievable, these locus numbers are nevertheless higher than the number of loci currently commonly applied in population genetic studies. The method of STRUCTURE seemed to be less sensitive to the proportion of hybrids included in the sample, while NEWHYBRIDS seemed to perform slightly better when individuals from both backcross and F1 hybrid classes were present in the sample. However, separating backcrosses from purebred parental individuals requires a considerable genotyping effort (at least 48 loci), even when divergence between parental populations is high. PMID- 16367831 TI - Natal philopatry does not lead to population genetic differentiation in Buller's albatross (Thalassarche bulleri bulleri). AB - Genetic variability in the only two existing populations of Buller's albatross (Thalassarche bulleri bulleri) was assessed using six polymorphic microsatellite loci. Large biological samples were obtained from both the Snares (n = 99) and the Solander Islands (n = 27). Several measures of genetic differentiation including F(ST) and R(ST) and a principal coordinates analysis (PCO) suggest a complete absence of genetic structure between three breeding colonies on the Snares Islands, and between them and one breeding colony on the Solander Islands. Mark/recapture studies of recently banded albatross chicks on the Snares found high natal philopatry in T. b. bulleri, but the microsatellite DNA data suggest that sufficient gene flow still occurs between all four breeding colonies to maintain a genetically homogeneous population overall. PMID- 16367832 TI - Estimating population size of endangered brush-tailed rock-wallaby (Petrogale penicillata) colonies using faecal DNA. AB - The brush-tailed rock-wallaby (Petrogale penicillata) is an endangered species in southeastern Australia and many of the remaining populations are declining. The steep rocky habitat and shy nature of the species make it difficult to obtain data on population parameters such as abundance and recruitment. Faecal pellet counts from scat plots are commonly used to monitor population trends but these are imprecise and difficult to relate to absolute population size. We conducted a noninvasive genetic sampling 'mark-recapture' study over a 2-year period to identify individuals from faecal DNA samples and estimate the population size of four brush-tailed rock-wallaby colonies located in Wollemi National Park, New South Wales. Scat plots in rock-wallaby colonies were used as sample collection points for this study. Two separate population estimates were carried out for three of the colonies to determine if we could detect recruitment and changes in population size. We determined that there was one large colony of an estimated 67 individuals (95% confidence interval: 55-91) and three smaller colonies. Monitoring of the smaller colonies also detected possible population size increases in all three. Our results indicate that faecal DNA analysis may be a promising method for estimating and monitoring population trends in this species particularly when used with a traditional field survey method. PMID- 16367833 TI - Population structure of brush-tailed rock-wallaby (Petrogale penicillata) colonies inferred from analysis of faecal DNA. AB - Genetic data obtained using faecal DNA were used to elucidate the population structure of four brush-tailed rock-wallaby (Petrogale penicillata) colonies located in Wollemi National Park, New South Wales. The results suggested that the four sampled colonies are genetically differentiated and do not form a panmictic unit. Based on assignment tests, approximately 5% of sampled individuals were inferred to be dispersers and both male and female migrants were detected. Multilocus spatial autocorrelation analyses provided evidence for increased philopatry among females compared to males within the largest colony in the valley. Females in close spatial proximity were more genetically similar than expected under a random distribution of females, and females separated by more than 400 m were less genetically similar than expected. In contrast, there was no evidence of a significant clustering of related males. This suggests that within colony dispersal is male biased. We also investigated the best strategies for conserving genetic diversity in this population. All of the four sampled colonies were found to contain distinct components of the genetic diversity of the Wolgan Valley P. penicillata population and loss of any colony is likely to result in the loss of unique alleles. Conservation and management plans should take into account that these colonies represent genetically differentiated discrete subpopulations. This approach is also the best strategy for maintaining the genetic diversity of the populations in this valley. PMID- 16367834 TI - Microsatellites reveal a lack of structure in Australian populations of the diamondback moth, Plutella xylostella (L.). AB - The diamondback moth, Plutella xylostella, is renowned for developing resistance to insecticides and causing significant economic damage to Brassica vegetable crops throughout the world. Yet despite its economic importance, little is known about the population structure and movement patterns of this pest both at local and regional scales. In Australia, the movement patterns and insecticide resistance status of P. xylostella infesting canola, vegetables, forage brassicas and weeds have fundamental implications for the management of this pest. Here we use six polymorphic microsatellite loci to investigate population structure and gene flow in Australian populations of P. xylostella. Samples of P. xylostella from New Zealand, Malaysia, Indonesia and Kenya were also scored at these loci. We found no evidence of population structure within Australia, with most populations having low inbreeding coefficients and in Hardy-Weinberg equilibrium. In addition, a sample from the North Island of New Zealand was indistinguishable from the Australian samples. However, large genetic differences were found between the Australia/New Zealand samples and samples from Kenya, Malaysia and Indonesia. There was no relationship between genetic distance and geographic distance among Australian and New Zealand samples. Two of the loci were found to have null alleles, the frequency of which was increased in the populations outside the Australia/New Zealand region. We discuss these results with reference to insecticide resistance management strategies for P. xylostella in Australia. PMID- 16367835 TI - Bayesian analyses of admixture in wild and domestic cats (Felis silvestris) using linked microsatellite loci. AB - Methods recently developed to infer population structure and admixture mostly use individual genotypes described by unlinked neutral markers. However, Hardy Weinberg and linkage disequilibria among independent markers decline rapidly with admixture time, and the admixture signals could be lost in a few generations. In this study, we aimed to describe genetic admixture in 182 European wild and domestic cats (Felis silvestris), which hybridize sporadically in Italy and extensively in Hungary. Cats were genotyped at 27 microsatellites, including 21 linked loci mapping on five distinct feline linkage groups. Genotypes were analysed with structure 2.1, a Bayesian procedure designed to model admixture linkage disequilibrium, which promises to assess efficiently older admixture events using tightly linked markers. Results showed that domestic and wild cats sampled in Italy were split into two distinct clusters with average proportions of membership Q > 0.90, congruent with prior morphological identifications. In contrast, free-living cats sampled in Hungary were assigned partly to the domestic and the wild cat clusters, with Q < 0.50. Admixture analyses of individual genotypes identified, respectively, 5/61 (8%), and 16-20/65 (25-31%) hybrids among the Italian wildcats and Hungarian free-living cats. Similar results were obtained in the past using unlinked loci, although the new linked markers identified additional admixed wildcats in Italy. Linkage analyses confirm that hybridization is limited in Italian, but widespread in Hungarian wildcats, a population that is threatened by cross-breeding with free-ranging domestic cats. The total panel of 27 loci performed better than the linked loci alone in the identification of domestic and known hybrid cats, suggesting that a large number of linked plus unlinked markers can improve the results of admixture analyses. Inferred recombination events led to identify the population of origin of chromosomal segments, suggesting that admixture mapping experiments can be designed also in wild populations. PMID- 16367836 TI - Hybrid origin and differentiation of two tetraploid Achillea species in East Asia: molecular, morphological and ecogeographical evidence. AB - Achillea (Asteraceae-Anthemideae) offers classical models for speciation by hybridization and polyploidy. Here, we test the suspected allotetraploid origin of two species, Achillea alpina and Achillea wilsoniana between phylogenetically distinct lineages in East Asia. A total of 421 AFLP bands from 169 individuals and 19 populations of five 2x- and two 4x-species were obtained. The data set was analysed with a newly developed model that accounts for polyploidy and assumes lack of recombination between the parental chromosome sets (i.e. disomic inheritance). A. alpina and A. wilsoniana then appear to be allotetraploids between Achillea acuminata-2x (sect. Ptarmica) and Achillea asiatica-2x (sect. Achillea). The two 4x-species share 44% and 48% of their AFLP bands with A. acuminata-2x, and 39% and 38% with A. asiatica-2x, respectively. Eight plastid haplotypes (A-H) were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses. A. alpina-4x and A. wilsoniana-4x share haplotype F only with A. asiatica-2x. This is consistent with the hybrid origin(s) involving the latter as the maternal ancestor. This result corroborates our previous DNA sequence data, where A. alpina-4x and A. wilsoniana-4x are also placed close to A. asiatica-2x. Morphology, ecology, and amplified fragment length polymorphism (AFLP) profiles of the two 2x-species are distinct, whereas the two 4x-species, grouped as A. alpina aggregate, form a nearly continuous link between them. Considering all evidence, this 4x-aggregate is regarded as the product of a hybridization between genetically distant 2x-ancestors limited to China and adjacent areas: one A. acuminata-like, and the other A. asiatica-like. The allopolyploid A. alpina agg. exhibits considerable morphological variation and ecological flexibility, and has expanded throughout eastern Asia and to northern North America, far beyond the ranges of their presumed 2x-ancestors. PMID- 16367837 TI - Effects of historical climate change, habitat connectivity, and vicariance on genetic structure and diversity across the range of the red tree vole (Phenacomys longicaudus) in the Pacific Northwestern United States. AB - Phylogeographical analyses conducted in the Pacific Northwestern United States have often revealed concordant patterns of genetic diversity among taxa. These studies demonstrate distinct North/South genetic discontinuities that have been attributed to Pleistocene glaciation. We examined phylogeographical patterns of red tree voles (Phenacomys longicaudus) in western Oregon by analysing mitochondrial control region sequences for 169 individuals from 18 areas across the species' range. Cytochrome b sequences were also analysed from a subset of our samples to confirm the presence of major haplotype groups. Phylogenetic network analyses suggested the presence of two haplotype groups corresponding to northern and southern regions of P. longicaudus' range. Spatial genetic analyses (samova and Genetic Landscape Shapes) of control region sequences demonstrated a primary genetic discontinuity separating northern and southern sampling areas, while a secondary discontinuity separated northern sampling areas into eastern and western groups divided by the Willamette Valley. The North/South discontinuity likely corresponds to a region of secondary contact between lineages rather than an overt barrier. Although the Cordilleran ice sheet (maximum approximately 12,000 years ago) did not move southward to directly affect the region occupied by P. longicaudus, climate change during glaciation fragmented the forest landscape that it inhabits. Signatures of historical fragmentation were reflected by positive associations between latitude and variables such as Tajima's D and patterns associated with location-specific alleles. Genetic distances between southern sampling areas were smaller, suggesting that forest fragmentation was reduced in southern vs. northern regions. PMID- 16367838 TI - Conflicting patterns of mitochondrial and nuclear DNA diversity in Phylloscopus warblers. AB - Molecular variation is often used to infer the demographic history of species, but sometimes the complexity of species history can make such inference difficult. The willow warbler, Phylloscopus trochilus, shows substantially less geographical variation than the chiffchaff, Phylloscopus collybita, both in morphology and in mitochondrial DNA (mtDNA) divergence. We therefore predicted that the willow warbler should harbour less nuclear DNA diversity than the chiffchaff. We analysed sequence data obtained from multiple samples of willow warblers and chiffchaffs for the mtDNA cytochrome b gene and four nuclear genes. We confirmed that the mtDNA diversity among willow warblers is low (pi = 0.0021). Sequence data from three nuclear genes (CHD-Z, AFLP-WW1 and MC1R) not linked to the mitochondria demonstrated unexpectedly high nucleotide diversity (pi values of 0.0172, 0.0141 and 0.0038) in the willow warbler, on average higher than the nucleotide diversity for the chiffchaff (pi values of 0.0025, 0.0017 and 0.0139). In willow warblers, Tajima's D analyses showed that the mtDNA diversity, but not the nuclear DNA diversity, has been reduced relative to the neutral expectation of molecular evolution, suggesting the action of a selective sweep affecting the maternally inherited genes. The large nuclear diversity seen within willow warblers is not compatible with processes of neutral evolution occurring in a population with a constant population size, unless the long-term effective population size has been very large (N(e) > 10(6)). We suggest that the contrasting patterns of genetic diversity in the willow warbler may reflect a more complex evolutionary history, possibly including historical demographic fluctuations or historical male-biased introgression of nuclear genes from a differentiated population of Phylloscopus warblers. PMID- 16367839 TI - European colonization by the spined loach (Cobitis taenia) from Ponto-Caspian refugia based on mitochondrial DNA variation. AB - In the last 20 years, new species, asexual reproduction, polyploidy and hybridization have all been reported within the genus Cobitis. An understanding of the current distribution and baseline phylogeographical history of 'true' nonhybrid Cobitis species is crucial in order to unravel these discoveries. In the present work, we investigated the phylogeography of the spined loach, Cobitis taenia, using 1126 bp of the mitochondrial cytochrome b gene from 174 individuals collected at 47 sites. In total, 51 haplotypes that differed at 49 positions (4.35%) were detected. We deduce that C. taenia survived European glaciations in at least three refugees in the Ponto-Caspian area. Two of these refugees each provided a major lineage that recolonized Europe in separate directions: one westward to England and the other spreading north into Russia before moving west. A third (minor) lineage that contributed little to the recolonization of Europe was also revealed--remaining near its Black Sea refuge. However, more recent history was difficult to resolve with colonization from a more western refugium during the last glacial maximum (LGM) a distinct possibility. Nested clade analysis indicates a pattern of restricted gene flow with isolation by distance at the first two levels and overall. Unlike many other European freshwater fish species, the Danube is not part of the current distribution of C. taenia, nor was it used as either a refuge or a source of colonization of Europe. Low genetic diversity within C. taenia suggests that its colonization of Europe is relatively recent. Demographic analyses revealed a history of recent expansion and isolation by distance. PMID- 16367840 TI - Gene lineages and eastern North American palaeodrainage basins: phylogeography and speciation in salamanders of the Eurycea bislineata species complex. AB - Contemporary North American drainage basins are composites of formerly isolated drainages, suggesting that fragmentation and fusion of palaeodrainage systems may have been an important factor generating current patterns of genetic and species diversity in stream-associated organisms. Here, we combine traditional molecular phylogenetic, multiple-regression, nested clade, and molecular-demographic analyses to investigate the relationship between phylogeographic variation and the hydrogeological history of eastern North American drainage basins in semiaquatic plethodontid salamanders of the Eurycea bislineata species complex. Four hundred forty-two sequences representing 1108 aligned bases from the mitochondrial genome are reported for the five formally recognized species of the E. bislineata complex and three outgroup taxa. Within the in-group, 270 haplotypes are recovered from 144 sampling locations. Geographic patterns of mtDNA-haplotype coalescence identify 13 putatively independent population-level lineages, suggesting that the current taxonomy of the group underestimates species-level diversity. Spatial and temporal patterns of phylogeographic divergence are strongly associated with historical rather than modern drainage connections, indicating that shifts in major drainage patterns played a pivotal role in the allopatric fragmentation of populations and build-up of lineage diversity in these stream-associated salamanders. More generally, our molecular genetic results corroborate geological and faunistic evidence suggesting that palaeodrainage connections altered by glacial advances and headwater erosion occurring between the mid-Miocene and Pleistocene epochs explain regional patterns of biodiversity in eastern North American streams. PMID- 16367841 TI - Comparative phylogeographic summary statistics for testing simultaneous vicariance. AB - Testing for simultaneous vicariance across comparative phylogeographic data sets is a notoriously difficult problem hindered by mutational variance, the coalescent variance, and variability across pairs of sister taxa in parameters that affect genetic divergence. We simulate vicariance to characterize the behaviour of several commonly used summary statistics across a range of divergence times, and to characterize this behaviour in comparative phylogeographic datasets having multiple taxon-pairs. We found Tajima's D to be relatively uncorrelated with other summary statistics across divergence times, and using simple hypothesis testing of simultaneous vicariance given variable population sizes, we counter-intuitively found that the variance across taxon pairs in Nei and Li's net nucleotide divergence (pi(net)), a common measure of population divergence, is often inferior to using the variance in Tajima's D across taxon pairs as a test statistic to distinguish ancient simultaneous vicariance from variable vicariance histories. The opposite and more intuitive pattern is found for testing more recent simultaneous vicariance, and overall we found that depending on the timing of vicariance, one of these two test statistics can achieve high statistical power for rejecting simultaneous vicariance, given a reasonable number of intron loci (> 5 loci, 400 bp) and a range of conditions. These results suggest that components of these two composite summary statistics should be used in future simulation-based methods which can simultaneously use a pool of summary statistics to test comparative the phylogeographic hypotheses we consider here. PMID- 16367842 TI - Beringian origins and cryptic speciation events in the fly agaric (Amanita muscaria). AB - Amanita muscaria sensu lato has a wide geographic distribution, occurring in Europe, Asia, Africa, Australia, New Zealand, and North, Central and South America. Previous phylogenetic work by others indicates three geographic clades (i.e. 'Eurasian', 'Eurasian-alpine' and 'North American' groups) within A. muscaria. However, the historical dispersal patterns of A. muscaria remained unclear. In our project, we collected specimens from arctic, boreal and humid temperate regions in Alaska, and generated DNA sequence data from the protein coding beta-tubulin gene and the internal transcribed spacer (ITS) and large subunit (LSU) regions of the ribosomal DNA repeat. Homologous sequences from additional A. muscaria isolates were downloaded from GenBank. We conducted phylogenetic and nested clade analyses (NCA) to reveal the phylogeographic history of the species complex. Although phylogenetic analyses confirmed the existence of the three above-mentioned clades, representatives of all three groups were found to occur sympatrically in Alaska, suggesting that they represent cryptic phylogenetic species with partially overlapping geographic distributions rather than being allopatric populations. All phylogenetic species share at least two morphological varieties with other species, suggesting ancestral polymorphism in pileus and wart colour pre-dating their speciations. The ancestral population of A. muscaria likely evolved in the Siberian-Beringian region and underwent fragmentation as inferred from NCA and the coalescent analyses. The data suggest that these populations later evolved into species, expanded their range in North America and Eurasia. In addition to range expansions, populations of all three species remained in Beringia and adapted to the cooling climate. PMID- 16367843 TI - A tale of two siblings: multiple paternity in big brown bats (Eptesicus fuscus) demonstrated using microsatellite markers. AB - In many bat species, the opportunity for sperm competition or other mechanisms of post-copulatory paternity biasing is thought to be great, due to the long delay between copulation and fertilization, demonstrated sperm storage capabilities, and observed promiscuity. We present the results of the first study to assess whether litters of big brown bats (Eptesicus fuscus) containing dizygotic twins share the same father. We sampled 26 mother-offspring triads from three colonies in Indiana and Illinois, as well as 299 additional adults (237 females and 62 males) from these colonies and six other maternity colonies in the same area in 1997-1998. All individuals were genotyped at nine highly variable autosomal microsatellite loci and one X-linked locus. We assessed multiple paternity using autosomal and X-linked locus exclusions, and using maximum-likelihood methods. All methods confirmed multiple paternity within litters, and the maximum likelihood analyses indicated that almost half of the sampled litters were composed of maternal half-siblings rather than full-siblings. Our results highlight the potential importance of post-copulatory mechanisms of paternity determination in the mating system of big brown bats, and have important implications for gene flow and population structuring in this species. PMID- 16367844 TI - Selective acquisition of novel mating type and vegetative incompatibility genes via interspecies gene transfer in the globally invading eukaryote Ophiostoma novo ulmi. AB - The Dutch elm disease fungus Ophiostoma novo-ulmi, which has destroyed billions of elm trees worldwide, originally invaded Europe as a series of clonal populations with a single mating type (MAT-2) and a single vegetative incompatibility (vic) type. The populations then rapidly became diverse with the appearance of the MAT-1 type and many vegetative incompatibility types. Here, we have investigated the mechanism using isolates from sites in Portugal at which the rapid evolution of O. novo-ulmi populations from clonality to heterogeneity was well established. We show by genetic mapping of vic and MAT loci with AFLP markers and by sequence analysis of MAT loci that this diversification was due to selective acquisition by O. novo-ulmi of the MAT-1 and vic loci from another species, Ophiostoma ulmi. A global survey showed that interspecies transfer of the MAT-1 locus occurred on many occasions as O. novo-ulmi spread across the world. We discuss the possibility that fixation of the MAT-1 and vic loci occurred in response to spread of deleterious viruses in the originally clonal populations. The process demonstrates the potential of interspecies gene transfer for facilitating rapid adaptation of invasive organisms to a new environment. PMID- 16367845 TI - Molecular evidence for a founder effect in invasive house finch (Carpodacus mexicanus) populations experiencing an emergent disease epidemic. AB - The impact of founder events on levels of genetic variation in natural populations remains a topic of significant interest. Well-documented introductions provide a valuable opportunity to examine how founder events influence genetic diversity in invasive species. House finches (Carpodacus mexicanus) are passerine birds native to western North America, with the large eastern North American population derived from a small number of captive individuals released in the 1940s. Previous comparisons using amplified fragment length polymorphism (AFLP) markers found equivalent levels of diversity in eastern and western populations, suggesting that any genetic effects of the founder event were ameliorated by the rapid growth of the newly established population. We used an alternative marker system, 10 highly polymorphic microsatellites, to compare levels of genetic diversity between four native and five introduced house finch populations. In contrast to the AFLP comparisons, we found significantly lower allelic richness and heterozygosity in introduced populations across all loci. Three out of five introduced populations showed significant reductions in the ratio of the number of alleles to the allele size range, a within-population characteristic of recent bottlenecks. Finally, native and introduced populations showed significant pairwise differences in allele frequencies in every case, with stronger isolation by distance within the introduced than native range. Overall, our results provide compelling molecular evidence for a founder effect during the introduction of eastern house finches that reduced diversity levels at polymorphic microsatellite loci and may have contributed to the emergence of the Mycoplasma epidemic which recently swept the eastern range of this species. PMID- 16367846 TI - Does disturbance enhance genotypic diversity in clonal organisms? A field test in the marine angiosperm Zostera marina. AB - Physical disturbance has often been invoked to control genotypic diversity in sessile clonal organisms, yet experimental evidence is lacking. I studied the effects of physical disturbance on genet dynamics and genotypic diversity in a clonal marine angiosperm, Zostera marina (eelgrass). In replicated plots of 1 m2, the vegetation canopy was removed in gaps of zero (control), 25%, 50% and 75% of the area (n = 6 replicates). Before removal and during two consecutive years, the genotypic composition was determined using genetic markers (DNA microsatellites) in a 5 x 5 pixel grid per plot. An aggregate index of genet dynamics summarizing recruitment, increase, loss and decrease of clones was maximal at intermediate disturbance levels (quadratic polynomial P = 0.02). Physical disturbance also increased the occurrence of new genotypes, possibly reflecting recruitment (linear model, P < 0.05). Contrary to expectations, there was no competitive advantage of more heterozygous genotypes over less heterozygous ones. In the absence of disturbance, in particular, clones with lower individual heterozygosity were more likely to increase in area over a 1-year time period than more heterozygous ones, while there was no such correlation in plots with disturbance (logistic model, P(disturbance x heterozygosity) = 0.036). Undisturbed plots revealed background recruitment independent of canopy gaps, suggesting that Z. marina exhibits a strategy of continual recruitment. Effects of experimental disturbance (linear or quadratic) on clonal diversity were not detectable. Instead, initial (pretreatment) clonal diversity accounted for between 68% and 91% of the variance in diversity, indicating remarkable resilience of genotypic diversity in the face of physical disturbance. PMID- 16367847 TI - Matching the origin of an invasive weed for selection of a herbivore haplotype for a biological control programme. AB - The Florida Everglades have been invaded by an exotic weed fern, Lygodium microphyllum. Across its native distribution in the Old World tropics from Africa to Australasia it was found to have multiple location-specific haplotypes. Within this distribution, the climbing fern is attacked by a phytophagous mite, Floracarus perrepae, also with multiple haplotypes. The genetic relationship between mite and fern haplotypes was matched by an overarching geographical relationship between the two. Further, mites that occur in the same location as a particular fern haplotype were better able to utilize the fern than mites from more distant locations. From a biological control context, we are able to show that the weed fern in the Everglades most likely originated in northern Queensland, Australia/Papua New Guinea and that the mite from northern Queensland offers the greatest prospect for control. PMID- 16367849 TI - A review of the effects of stannous fluoride on gingivitis. AB - AIM: To review the literature on the effects of stannous fluoride on gingivitis. MATERIAL AND METHODS: The Medline and cochrane central register of controlled trials were searched up to August 2005 to identify appropriate studies. The primary outcome measure was gingivitis. RESULTS: Independent screening of titles and abstracts of 542 papers resulted in 36 publications (inter-reviewer e score of 0.76), out of which 15 papers finally fulfilled the criteria of eligibility. For SnF(2)dentifrices, a statistically significant reduction in gingivitis was noted in comparison with control (weighted mean difference (WMD) of 0.15 (gingival index) and 0.21 (gingivitis severity index) (test for heterogeneity p<0.00001, I(2)=91.1% and p=0.03, I(2)=80.1%, respectively)). With regard to plaque reduction inconsistent results existed. On using the plaque index no differences were found, whereas meta-analysis of the Turesky index provided a WMD of 0.31 (p=0.01, test for heterogeneity p<0.0001, I(2)=91.7%). Because of insufficient data, a meta-analysis for SnF(2)mouth rinse and dentifrice/mouthrinse formulations was not performed. CONCLUSIONS: The use of SnF(2) dentifrices results in gingivitis and plaque reduction when compared with a conventional dentifrice. The precise magnitude of this effect was difficult to assess because of a high level of heterogeneity in study outcomes. PMID- 16367850 TI - A method to study sustained antimicrobial activity of rinse and dentifrice components on biofilm viability in vivo. AB - AIM: To develop an improved method for quantitative assessment of antimicrobial efficacy and substantivity of mouth rinses and dentifrices on in vivo treated plaque. MATERIAL AND METHODS: Nine- and 72-h-old plaques were formed in volunteers carrying out standardized hygiene using NaF-containing dentifrice. Plaques were collected before (baseline) in vivo treatment with dentifrices or chlorhexidine mouth rinse, immediately post-treatment and after 1 or 6 h, dispersed in demineralized water and stained with live/dead stain after which bacteria were enumerated. Dispersed baseline plaques were treated with dentifrices or chlorhexidine to determine antimicrobial efficacy against planktonic bacteria. RESULTS: Baseline plaques revealed 56-41% viable organisms in 9- and 72-h-old plaques, respectively. Treatment of planktonic (dispersed baseline plaque) bacteria resulted in 1-4% viable organisms. Chlorhexidine mouth rinse and dentifrices produced strong immediate antimicrobial effects, but after 1 or 6 h, the proportion of viable organisms in 9-h-old plaques rebounded significantly with only chlorhexidine mouth rinse retaining significant efficacy. Seventy-two-hour-old plaques were less susceptible to antimicrobials, although dentifrices appeared more effective after 6 h than initially, whereas efficacy of chlorhexidine rinse continued to drop with time post-treatment. CONCLUSIONS: The proposed method holds promise for assessment of both immediate and retained antimicrobial actions of oral treatments against dental plaque in vivo. PMID- 16367851 TI - Cynical hostility as a determinant of toothbrushing frequency and oral hygiene. AB - AIM: Our aim was to investigate whether cynical hostility, self-reported toothbrushing frequency and objectively assessed levels of oral hygiene were associated. MATERIAL AND METHODS: The present study sample consisted of 4156 30 64-year-old dentate Finns. The questionnaire and the home interview included information about socioeconomic and sociodemographic factors and behavioural variables, such as toothbrushing frequency, dental attendance, smoking and cynical hostility. The level of oral hygiene was assessed during a clinical oral examination. The chi(2) test and ordinal logistic regression analyses were used. RESULTS: After controlling for sociodemographic and socioeconomic factors, smoking habits and reported dental attendance, the subjects belonging to the lowest cynical hostility level were found to brush their teeth significantly more often and to have better oral hygiene compared with those belonging the highest cynical hostility level. Toothbrushing frequency was adjusted when oral hygiene was used as the outcome variable. The association of cynical hostility with toothbrushing frequency and the oral hygiene seems to be partly dependent on the level of education. CONCLUSIONS: Cynical hostility is a psychosocial risk marker for the frequency and quality of toothbrushing and it could be a connecting trait between general health behaviour and oral health behaviour. PMID- 16367852 TI - Bad breath and periodontal disease: how related are they? PMID- 16367853 TI - Oral malodor-related parameters in the Chinese general population. AB - OBJECTIVES: To estimate the prevalence of halitosis in the Chinese population and to assess the relationships between halitosis and oral health, social and behavioural factors. METHODS: The correlation between the incidence of oral malodor and oral health was surveyed in a sample of 2000 individuals (1000 males and 1000 females) aged 15-64 years residing in urban and rural areas. Malodor was measured with both organoleptic measurements and with a portable sulphide monitor. Assessment of oral health included decayed, missing and filled teeth (DMFT), periodontal status, dental plaque, and tongue coating. Behavioural and social factors related with oral health or halitosis were also investigated. RESULTS: The prevalence of halitosis was 27.5% according to the organoleptic score. The level of volatile sulphur compounds (VSCs) in mouth air was significantly lower in males and in some of the age groups after lunch. Age and location of residence (rural or urban areas) did not influence the VSCs concentration in mouth air. The amount of tongue coating played the most important role in increasing VSCs concentration in mouth air, followed by periodontal status and plaque index values. DMFT, social, and behavioural factors did not contribute to halitosis. CONCLUSIONS: Tongue coating score, modified sulcus bleeding index and calculus index were factors significantly related to oral malodor in this study. PMID- 16367854 TI - The effect of wear on ultrasonic scaler tip displacement amplitude. AB - AIM: During clinical usage, scaler tips may become worn and reduced in length. It is unknown what effect wear has on the magnitude of scaler tip vibrations when they are utilized under typical clinical loads. The aim of this investigation was to assess the effect of simulated wear on ultrasonic scaler tip displacement amplitude, using a scanning laser vibrometer. MATERIALS AND METHODS: A Cavitron SPS (Dentsply) ultrasonic generator and three scaler insert designs (FSI-100, FSI 1000 and FSI-SLI-10S) were selected for the investigation. Tip vibration displacement amplitude was assessed unloaded and then contacting against tooth surfaces with loads of 0.5 and 1.0 N. Tips were then ground down by 1 mm and then 2 mm and scans were repeated. RESULTS: For all tips, load and length were found to be significant variables (p<0.0001). The scaler tips showed a fall in displacement amplitude with a reduction in tip length. However, all scaler tips showed variability in the amount oscillation that occurred. This was most pronounced with FSI-SLI-10S. CONCLUSIONS: This investigation demonstrated that tip wear could affect the performance of dental ultrasonic scaler inserts by reducing their vibration displacement amplitude. Clinicians should be aware of this variability, which may be significant enough to affect clinical procedures. PMID- 16367855 TI - Immediate effect of instrumentation on the subgingival microflora in deep inflamed pockets under strict plaque control. AB - OBJECTIVE: To investigate (1) reduction in the number of microorganisms obtained directly after subgingival instrumentation, (2) rate of bacterial re-colonization during 2 weeks, under supragingival plaque-free conditions. MATERIALS AND METHODS: Effects of subgingival instrumentation were measured at one deep pocket in 22 patients (11 smokers and 11 non-smokers). Immediately after initial therapy, experimental sites, under strict plaque control, were instrumented subgingivally. Microbiological evaluation was performed at pre-instrumentation, immediate post-instrumentation and 1 and 2 weeks post-instrumentation. RESULTS: Mean total anaerobic colony forming units (CFUs) dropped from 3.9 x 10(6) before to 0.09 x 10(6) immediately following instrumentation. Significant reductions were found for Tannerella forsythia, Micromonas micros, Fusobacterium nucleatum and spirochetes. Significant reductions were not observed for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and Campylobacter rectus. Except for spirochetes, no reduction in prevalence of specific periodontal bacteria was found immediately after instrumentation. During follow-up, mean total CFU tended to increase. Prevalence of periodontal bacteria further reduced. No effect of smoking was found. CONCLUSION: Results indicate that subgingival mechanical cleaning in itself, has a limited effect, in actually removing bacteria. The subsequent reduction in prevalence of specific periodontal bacteria shows that it is apparently difficult for these species to survive in treated pockets. PMID- 16367856 TI - Impact of antiseptics on one-stage, full-mouth disinfection. PMID- 16367857 TI - Loss of molars in periodontally treated patients: a retrospective analysis five years or more after active periodontal treatment. AB - OBJECTIVES: Evaluation of tooth loss in molars and prognostic factors for molar survival. MATERIAL AND METHODS: Five hundred and five molars in 71 patients (mean age 46 years; 40 females) were evaluated. The following inclusion criteria were required: periodontal therapy of at least one molar, at least 5 years of supportive periodontal therapy, and baseline assessment of furcation involvement (FI). RESULTS: At baseline 200 of 505 molars exhibited no FI, 116 degree I, 122 degree II, and 67 degree III FI. Twenty-seven molars did not receive periodontal treatment; 127 molars were subjected to non-surgical therapy, and 227 to flap surgery. Tunnel preparation was performed on 14 molars, root resection on 20, regenerative therapy on 57 teeth, and 33 molars were extracted. During the average follow-up period of 107 months 38 molars were lost additionally. Molars with degree III FI had the highest mortality. A multi-level proportional hazard model revealed smoking, baseline bone loss, number of molars left, and degree III FI as risk factors influencing the retention time of molars. CONCLUSION: Overall periodontal therapy results in a good prognosis of molars. Degree III FI leads to a significant deterioration of prognosis. Beyond FI smoking, baseline bone loss, and number of molars left influence molar survival. PMID- 16367858 TI - Microbial colonization patterns predict the outcomes of surgical treatment of intrabony defects. AB - AIM: To explore the impact of bacterial load and microbial colonization patterns on the clinical outcomes of periodontal surgery at deep intrabony defects. MATERIALS AND METHODS: One hundred and twenty-two patients with advanced chronic periodontitis and at least one intrabony defect of >3 mm were recruited in 10 centres. Before recruitment, the infection control phase of periodontal therapy was completed. After surgical access and debridement, the regenerative material was applied in the test subjects, and omitted in the controls. At baseline and 1 year following the interventions, clinical attachment levels (CAL), pocket probing depths (PPD), recession (REC), full-mouth plaque scores and full-mouth bleeding scores were assessed. Microbial colonization of the defect-associated pocket was assessed using a DNA-DNA checkerboard analysis. RESULTS: Total bacterial load and counts of red complex bacteria were negatively associated with CAL gains 1 year following treatment. The probability of achieving above median CAL gains (>3 mm) was significantly decreased by higher total bacterial counts, higher red complex and T. forsythensis counts immediately before surgery. CONCLUSIONS: Presence of high bacterial load and specific periodontal pathogen complexes in deep periodontal pockets associated with intrabony defects had a significant negative impact on the 1 year outcome of surgical/regenerative treatment. PMID- 16367859 TI - Autogenous bone graft in conjunction with enamel matrix derivative in the treatment of deep periodontal intra-osseous defects: a report of 13 consecutively treated patients. AB - AIM: The purpose of the present study was to investigate the effectiveness of a regenerative procedure based on supra-crestal soft tissue preservation in association with combined autogenous bone (AB) graft/enamel matrix derivative (EMD) application in the treatment of deep periodontal intra-osseous defects. METHODS: Thirteen consecutively treated patients, seven females and six males, aged 30-65 years, three smokers, were included. A total of 15 deep, one- to two wall intra-osseous defects were selected. Immediately before surgery and 6 months after surgery, pocket probing depth (PPD), clinical attachment level (CAL), and gingival recession (REC) were recorded. RESULTS: PPD amounted to 9.4+/-1.8 mm before surgery, and decreased to 4.7+/-1.2 mm post-surgery (p<0.0000). CAL varied from 10.5+/-2.0 mm pre-surgery to 6.2+/-1.7 mm post-surgery (p<0.0000), with CAL gain averaging 4.3+/-1.4 mm. Fourteen (93.3%) defects presented CAL gain >/=3 mm. REC change was 0.4+/-0.7 mm. CONCLUSIONS: Results from the present study indicated that a regenerative procedure based on supra-crestal soft tissue preservation and combined AB/EMD treatment leads to a clinically and statistically significant improvement of soft tissue conditions of deep periodontal intra-osseous defects. PMID- 16367860 TI - Soft tissue myxoma: report of an unusual case located on the gingiva. AB - BACKGROUND: Soft tissue myxoma of the oral cavity is rare. Only three cases of myxoma of gingiva have been reported in the literature. We present a case of soft tissue myxoma arising from the left maxillary adherent gingiva in a 42-year-old male [correction of female] patient. METHODS: Histological examination showed spindle-shaped cells in a myxoid stroma. Immunohistochemical stains with S-100 protein were negative, while those with vimentin were positive. RESULTS: Clinical examination revealed a soft tissue mass, with tense elastic consistency on palpation. The overlying mucosa was normal and healthy. A clinical diagnosis of fibroma was given. Histological examination showed spindle-shaped and stellate cells, arranged in a myxoid fibrous stroma, with collagen fibres distributed uniformly. Scattered islands or strands of inactive odontogenic epithelium were present. On the basis of the histological and immunohistochemical findings, the final diagnosis was soft tissue myxoma. CONCLUSIONS: Further studies are necessary to clarify the origin and histogenesis of this lesion. PMID- 16367861 TI - Contribution of host lipids to Toxoplasma pathogenesis. AB - As an actively dividing organism, the intracellular parasite Toxoplasma gondii must adjust the size and composition of its membranes in order to accommodate changes due to housekeeping activities, to commit division and in fine to produce new viable progenies. Lipid inventory of T. gondii reveals that the biological membranes of this parasite are composed of a complex mixture of neutral and polar lipids. After examination of the origin of T. gondii membrane lipids, three categories of lipids can be described: (i) lipids scavenged by T. gondii from the host cell; (ii) lipids synthesized in large amounts by the parasite, independently from its host cell; and (iii) lipids produced de novo by the parasite, but whose synthesis does not come close to satisfying the entire parasite's needs. These latter must be adeptly acquired from the host environment. To this end, T. gondii diverts a large variety of lipid precursors from host cytoplasm and efficiently manufacture them into complex lipids. This rather remarkable reliance on host lipid resources for parasite survival opens new avenues to restrict parasite growth. Indeed, parasite starvation can be induced upon deprivation from essential host lipids. Lipid analogues with anti proliferative properties are voraciously taken up by the parasites, which results in parasite membrane defects, and ultimately death. PMID- 16367862 TI - Carbon metabolism of intracellular bacteria. AB - Bacterial metabolism has been studied intensively since the first observations of these 'animalcules' by Leeuwenhoek and their isolation in pure cultures by Pasteur. Metabolic studies have traditionally focused on a small number of model organisms, primarily the Gram negative bacillus Escherichia coli, adapted to artificial culture conditions in the laboratory. Comparatively little is known about the physiology and metabolism of wild microorganisms living in their natural habitats. For approximately 500-1000 species of commensals and symbionts, and a smaller number of pathogenic bacteria, that habitat is the human body. Emerging evidence suggests that the metabolism of bacteria grown in vivo differs profoundly from their metabolism in axenic cultures. PMID- 16367863 TI - Molecular motors hijacking by intracellular pathogens. AB - Cargoes are transported intracellularly along cytoskeletal tracks composed of actin or tubulin. Their movement involves the action of molecular motor proteins that generate directed movement along microtubules or actin filaments. The three classes of molecular motors--kinesins, dyneins and myosins--are involved in a multiplicity of biological movements such as mitosis, positioning of organelles, intracellular transports and also vesicular sorting through membrane tubulation and fission and delivery to their target compartment. Intracellular pathogens use this molecular machinery to reach their site of replication, to leave their host or to control the dynamics of membrane exchanges with their replication compartment. PMID- 16367864 TI - Magnetic resonance imaging of Klebsiella pneumoniae-induced pneumonia in mice. AB - In vivo imaging of small animals is a rapidly developing field. However, the potential of global imaging of infectious processes in animal models remains poorly explored. We used magnetic resonance imaging (MRI) to follow the development and regression of inflammatory lesions caused by infection by Klebsiella pneumoniae in mouse lungs. A virulent strain caused an intense inflammation within 2 days in the whole lungs, while an avirulent strain did not show significant changes. Mice infected with the virulent strain and subsequently treated with antibiotics presented a severe inflammation localized mainly in the left lung that disappeared after a week. The lesions observed by MRI correlated with the damage seen by histological analysis and a 3D representation of the tissue allowed better visualization of the development and healing of inflammatory lesions. MRI thus represents a powerful technique to study in vivo the interactions between a pathogen and its host in real time. PMID- 16367865 TI - Galectin-3 binds to Helicobacter pylori O-antigen: it is upregulated and rapidly secreted by gastric epithelial cells in response to H. pylori adhesion. AB - Helicobacter pylori causes gastritis and some infections result in peptic ulceration, gastric adenocarcinoma or gastric lymphoma. A critical step in the pathogenesis of these diseases is the ability of H. pylori to adhere to gastric epithelial cells. A role for the lipopolysaccharide O-antigen side-chain in this process has previously been identified. In this study, evidence is presented that the receptor recognized by the O-antigen side-chain is galectin-3, a beta galactoside-binding lectin. A variety of functions have been ascribed to galectin 3 including modulation of extracellular adhesion and chemotaxis of monocytes and neutrophils. Expression of galectin-3 is upregulated by gastric epithelial cells following adhesion of H. pylori, suggesting that in addition to colonization this protein also plays a role in the host response to infection. Upregulation of galectin-3 is inhibited by treating gastric epithelial cells with the mitogen activated protein kinase (MAPK) inhibitors U0126 or PD098059 and does not occur in cells infected with either H. pylori cagE or cagA isogenic mutants. This implies that H. pylori-mediated expression of galectin-3 is dependent on delivery of CagA into the host cell cytosol and the subsequent stimulation of MAPK signalling. A further consequence of H. pylori adhesion is that it elicits a rapid release of galectin-3 from infected cells. A role for this phenomenon in initiating the trafficking of phagocytic cells to the site of infection is discussed. PMID- 16367866 TI - Host protein interactions with enteropathogenic Escherichia coli (EPEC): 14-3 3tau binds Tir and has a role in EPEC-induced actin polymerization. AB - Enteropathogenic Escherichia coli (EPEC) cause infantile diarrhoea and are characterized by their ability to produce attaching and effacing lesions on the surface of intestinal epithelial cells. EPEC employ a filamentous type III secretion system to deliver effector molecules that subvert mammalian cell function to generate actin- and cytokeratin-rich pedestals beneath adherent bacteria. Tir is a major effector protein that is delivered to the plasma membrane of the eukaryotic cell where it acts as the receptor for the bacterial adhesin intimin. Host cell proteins that are recruited to the site of intimate attachment include focal adhesion and cytoskeletal proteins that contribute to pedestal formation. We have used Tir as bait in a yeast two-hybrid screen to identify the protein 14-3-3tau as a binding partner. 14-3-3 proteins are a family of adaptor proteins that modulate protein function in all eukaryotic cells. Here we demonstrate that the tau isoform (also known as theta) of 14-3-3 can bind specifically to Tir in a phosphorylation-independent manner, and that the interaction occurs during the infection process by co-immunoprecipitation of the partners from infected HeLa cell extracts. 14-3-3tau is recruited to the site of the pedestal (3 h after infection) and can decorate attached EPEC in the later stages of the infection process (5-7 h). Pedestal formation can be impaired by depletion of cellular 14-3-3tau using small interfering RNAs. This study indicates a direct functional role for the 14-3-3tau:Tir interaction and is the first to demonstrate the association of a host protein with the surface of EPEC. PMID- 16367867 TI - Resistance of fluorescent-labelled Actinobacillus actinomycetemcomitans strains to phagocytosis and killing by human neutrophils. AB - Neutrophils are initially the predominant cells involved in the host defence of bacterial infections, including periodontal disease. Aggressive periodontitis is associated with Actinobacillus actinomycetemcomitans, a Gram-negative capnophilic microorganism. Infections caused by A. actinomycetemcomitans are not resolved by the host immune response despite the accumulation of neutrophils at the site of inflammation. To better understand the role of natural host defence mechanisms in A. actinomycetemcomitans infections, the interaction of phenotypically diverse strains of this pathogen with human neutrophils was assessed directly using techniques such as genetic labelling with the gene for green fluorescent protein, fluorescence-activated cell sorting and fluorescence imaging. The study included clinical isolates of A. actinomycetemcomitans represented by self-aggregating, biofilm-associated and isogenic planktonic variants. Data obtained showed that complement-mediated phagocytosis of A. actinomycetemcomitans was generally inefficient regardless of strain-specific serotype or leukotoxin production. Furthermore, the majority of ingested bacteria remained viable after exposure to neutrophils for 1 h. Interestingly, uptake of antibody-opsonized bacteria resulted in the rapid cell death of neutrophils. This was in contrast to ingestion of complement-opsonized bacteria, which did not affect neutrophil viability. The methods used in this study provided reliable and reproducible results with respect to adherence, phagocytosis and killing of A. actinomycetemcomitans when encountering human neutrophils. PMID- 16367868 TI - Elevated mitogen-activated protein kinase signalling and increased macrophage activation in cells infected with a glycopeptidolipid-deficient Mycobacterium avium. AB - Mycobacterium avium is a significant cause of morbidity and mortality in AIDS patients. M. avium can be isolated as three major morphotypes: smooth-transparent (SmT ), smooth-opaque (SmO) and rough (Rg). Studies indicate that many Rg isolates lack or have modified glycopeptidolipids (GPLs). GPLs are major surface constituents of the M. avium cell wall and heterogeneity in their carbohydrate moieties has been used to classify M. avium into different serotypes, with serotypes 1, 4 and 8 being isolated with high frequency from AIDS patients. However, it is unclear what role GPLs play in M. avium pathogenicity. To begin to address how the absence of GPLs affects M. avium-macrophage interaction, we used the well-characterized M. avium 2151 SmT and Rg isolates which differ in GPL expression. We found macrophages infected with the Rg compared with SmT M. avium 2151 showed prolonged activation of the mitogen-activated protein kinases (MAPKs) p38 and ERK1/2. Macrophages infected with the Rg 2151 also showed increased tumour necrosis factor-alpha (TNF-alpha) production. Interestingly, TNF-alpha secretion by macrophages infected with SmO or SmT 2151 was dependent on p38, ERK1/2 and NF-kappaB while TNF-alpha secretion by Rg 2151-infected macrophages was dependent on NF-kappaB but not the MAPKs. Rg 2151-infected macrophages also produced increased levels of IL-6, IL-12, MCP-1 and RANTES relative to macrophages infected with SmT 2151. These results indicate that M. avium 2151 deficient in GPLs promote increased macrophage activation. This disparity in cellular activation stems from a quantitative and qualitative difference in the macrophage signalling response to the Rg and SmT M. avium 2151. PMID- 16367869 TI - Helicobacter pylori CagA transfection of gastric epithelial cells induces interleukin-8. AB - To determine the effect of Helicobacter pylori CagA expression on interleukin-8 (IL-8) induction in AGS cells, cagA and five of its fragments from strains 147A and 147C that vary in the 3' repeat region were cloned into the eukaryotic expression plasmid pSP65SRalpha. IL-8, but not RANTES or IL-Ibeta, levels were increased in AGS cells transfected with 147A-cagA and to a greater extent with 147C-cagA, compared with negative controls. The 5' b fragment from the two strains had similar effects, but the 3' d and e fragments from 147C CagA had greater effects than those from 147A-CagA. When the Western CagA-specific sequence (WSS) of 147C-cagA was replaced with East Asian CagA-specific sequence (ESS) and cloned into pSP65SRalpha as an East/West chimera, there was no significant effect on IL-8 production. Use of specific inhibitors indicates that Src kinase activation, and the mitogen-activated protein (MAP) kinase and NF kappaB pathways are the major intermediates for CagA effects on IL-8 induction, but the p38 MAP kinase pathway has little effect. These results indicate a direct CagA effect on IL-8 induction by gastric epithelial cells, and indicate signal pathway loci that can be targeted for amelioration. PMID- 16367870 TI - Cytolysin-dependent delay of vacuole maturation in macrophages infected with Listeria monocytogenes. AB - The bacterial pathogen Listeria monocytogenes (Lm) evades the antimicrobial mechanisms of macrophages by escaping from vacuoles to the cytosol, through the action of the cytolysin listeriolysin O (LLO). Because of heterogeneities in the timing and efficiency of escape, important questions about the contributions of LLO to Lm vacuole identity and trafficking have been inaccessible. Expression of cyan fluorescent protein (CFP)-labelled endocytic membrane markers in macrophages along with a yellow fluorescent protein (YFP)-labelled indicator of Lm entry to the cytosol identified compartments lysed by bacteria. Lm escaped from Rab5a negative, lysosome-associated membrane protein-1 (LAMP1)-negative, Rab7-positive, phosphatidylinositol 3-phosphate [PI(3)P]-positive vacuoles. Lm vacuoles did not label with YFP-Rab5a unless the bacteria were first opsonized with IgG. Wild-type Lm delayed vacuole fusion with LAMP1-positive lysosomes, relative to LLO deficient Lm. Bacteria prevented from expressing LLO until their arrival into LAMP1-positive lysosomes escaped inefficiently. Thus, the LLO-dependent delay of Lm vacuole fusion with lysosomes affords Lm a competitive edge against macrophage defences by providing bacteria more time in organelles they can penetrate. PMID- 16367871 TI - Expression of a Porphyromonas gingivalis lipid A palmitylacyltransferase in Escherichia coli yields a chimeric lipid A with altered ability to stimulate interleukin-8 secretion. AB - In Escherichia coli the gene htrB codes for an acyltransferase that catalyses the incorporation of laurate into lipopolysaccharide (LPS) as a lipid A substituent. We describe the cloning, expression and characterization of a Porphyromonas gingivalis htrB homologue. When the htrB homologue was expressed in wild-type E. coli or a mutant strain deficient in htrB, a chimeric LPS with altered lipid A structure was produced. Compared with wild-type E. coli lipid A, the new lipid A species contained a palmitate (C16) in the position normally occupied by laurate (C12) suggesting that the cloned gene performs the same function as E. coli htrB but preferentially transfers the longer-chain palmitic acid that is known to be present in P. gingivalis LPS. LPS was purified from wild-type E. coli, the E. coli htrB mutant strain and the htrB mutant strain expressing the P. gingivalis acyltransferase. LPS from the palmitate bearing chimeric LPS as well as the htrB mutant exhibited a reduced ability to activate human embryonic kidney 293 (HEK293) cells transfected with TLR4/MD2. LPS from the htrB mutant also had a greatly reduced ability to stimulate interleukin-8 (IL-8) secretion in both endothelial cells and monocytes. In contrast, the activity of LPS from the htrB mutant bacteria expressing the P. gingivalis gene displayed wild-type activity to stimulate IL-8 production from endothelial cells but a reduced ability to stimulate IL-8 secretion from monocytes. The intermediate activation observed in monocytes for the chimeric LPS was similar to the pattern seen in HEK293 cells expressing TLR4/MD2 and CD14. Thus, the presence of a longer-chain fatty acid on E. coli lipid A altered the activity of the LPS in monocytes but not endothelial cell assays and the difference in recognition does not appear to be related to differences in Toll-like receptor utilization. PMID- 16367872 TI - Inhibition of mitogen-activated protein kinase signalling by Bacillus anthracis lethal toxin causes destabilization of interleukin-8 mRNA. AB - Bacillus anthracis must overcome host innate immune defences to establish a systemic anthrax infection. This is facilitated in part by lethal toxin (LT), a secreted virulence factor that consists of a cell-binding moiety, protective antigen (PA), and an enzymatic moiety, lethal factor (LF). PA binds cells through protein receptors and mediates the delivery of LF to the cytosol. LF is a protease that cleaves amino-terminal fragments from mitogen-activated protein kinase kinases (MAPKKs), preventing phosphorylation of their downstream targets. Here we report that LT reduces the amount of interleukin (IL)-8 produced and secreted by human endothelial cells. The reduction of IL-8 levels by LT was not attributable to reduced expression from the IL-8 promoter, but resulted from destabilization of IL-8 mRNA. Destabilization by LT was mediated through the 3' untranslated region of the IL-8 transcript and could be mimicked by pharmacological inhibitors of MAPK pathways. LT diminished the induction of IL-8 mRNA and protein by lipopolysaccharide, indicating that the toxin can impair the ability of these cells to initiate an immune response. Destabilization of a cytokine transcript represents a new interference strategy used by either a bacterial or viral pathogen to reduce cytokine expression and may help B. anthracis to evade host immune defences. PMID- 16367873 TI - Specific host genes required for the killing of Klebsiella bacteria by phagocytes. AB - The amoeba Dictyostelium discoideum shares many traits with mammalian macrophages, in particular the ability to phagocytose and kill bacteria. In response, pathogenic bacteria use conserved mechanisms to fight amoebae and mammalian phagocytes. Here we developed an assay using Dictyostelium to monitor phagocyte-bacteria interactions. Genetic analysis revealed that the virulence of Klebsiella pneumoniae measured by this test is very similar to that observed in a mouse pneumonia model. Using this assay, two new host resistance genes (PHG1 and KIL1) were identified and shown to be involved in intracellular killing of K. pneumoniae by phagocytes. Phg1 is a member of the 9TM family of proteins, and Kil1 is a sulphotransferase. The loss of PHG1 resulted in Dictyostelium susceptibility to a small subset of bacterial species including K. pneumoniae. Remarkably, Drosophila mutants deficient for PHG1 also exhibited a specific susceptibility to K. pneumoniae infections. Systematic analysis of several additional Dictyostelium mutants created a two-dimensional virulence array, where the complex interactions between host and bacteria are visualized. PMID- 16367874 TI - Chlamydia trachomatis enters a viable but non-cultivable (persistent) state within herpes simplex virus type 2 (HSV-2) co-infected host cells. AB - Epidemiological and clinical studies have shown that double infection with herpes simplex virus type 2 (HSV-2) and Chlamydia trachomatis occurs in vivo. We hypothesized that co-infection would alter replication of these agents. To test this hypothesis, HeLa cells were infected with C. trachomatis serovar E, followed 24 h later by HSV-2 strain 333. Transmission electron microscopic (TEM) analyses indicated that, by 10 h after HSV addition, reticulate bodies (RBs) in co infected cells were swollen, aberrantly shaped and electron-lucent. In infectious titre assays, HSV-2 co-infection abrogated production of infectious chlamydial progeny. Western blot analyses indicated that accumulation of chlamydial major outer membrane protein (MOMP) was decreased by HSV co-infection while accumulation of chlamydial heat-shock protein 60-1 (HSP60-1) was increased. Polymerase chain reaction (PCR) experiments indicated that chlamydial genome copy number was unaltered by HSV-2 superinfection. Semi-quantitative, reverse transcription PCR (RT-PCR) experiments demonstrated that levels of chlamydial groEL, ftsK, ftsW, dnaA and unprocessed 16S rRNA transcripts were not changed by HSV-2 super-infection. These data indicate that HSV-2 superinfection drives chlamydia into a viable but non-cultivable state, which is the hallmark of persistence. Because chlamydial HSP60-1 has been associated with immunopathology in vivo, these results also suggest that disease severity might be increased in co-infected individuals. PMID- 16367875 TI - A mosquito-specific protein family includes candidate receptors for malaria sporozoite invasion of salivary glands. AB - We describe a previously unrecognized protein family from Aedes and Anopheles mosquitoes, here named SGS proteins. There are no SGS homologues in Drosophila or other eukaryotes, but SGS presence in two mosquito genera suggests that the protein family is widespread among mosquitoes. Ae. aegypti aaSGS1 mRNA and protein are salivary gland specific, and protein is localized in the basal lamina covering the anatomical regions that are preferentially invaded by malaria sporozoites. Anti-aaSGS1 antibodies inhibited sporozoite invasion into the salivary glands in vivo, confirming aaSGS1 as a candidate sporozoite receptor. By homology to aaSGS1 we identified the complete complement of four SGS genes in An. gambiae, which were not recognized in the genome annotation. Two An. gambiae SGS genes display salivary gland specific expression like aaSGS1. Bioinformatic analysis predicts that SGS proteins possess heparin-binding domains, and have among the highest density of tyrosine sulphation sites of all An. gambiae proteins. The major sporozoite surface proteins (CS and TRAP) also bind heparin, and interact with sulphoconjugates during liver cell invasion. Thus, we speculate that sporozoite invasion of mosquito salivary glands and subsequently the vertebrate liver may share similar mechanisms based on sulphation. Phylogenomic analysis suggests that an SGS ancestor was involved in a lateral gene transfer. PMID- 16367876 TI - Current indications for pancreas or islet transplant. AB - Pancreas or islet transplantation can provide good glycaemic control and insulin independence. Pancreas transplantation has been associated with improvement in diabetic retinopathy, nephropathy, neuropathy and vasculopathy, but has the associated morbidity of major surgery. Both forms of therapy require long-term immunosuppression and its attendant risks and both achieve insulin independence rates of about 80% at 1 year. Pancreas transplantation at the same time as a renal transplant is a worthwhile option to employ, especially if the diabetes has been difficult to control. Diabetes associated with frequent severe hypoglycaemia or extreme lability, despite optimization of diabetes management, may benefit from either pancreas or islet transplant alone with the latter being the lower risk procedure. More quantitative measures of hypoglycaemia and lability are now available to facilitate the assessment of the severity of these problems with glucose control. Diabetic patients with renal involvement (macroproteinuria, but no major elevation of creatinine) and unstable diabetes may be helped with an islet or pancreas transplant, but this approach should still be considered experimental and such a transplant may hasten the need for renal replacement therapy. In the setting of well-controlled diabetes and intact renal function, it is difficult to justify pancreas or islet transplant alone given the risks of immunosuppression. PMID- 16367877 TI - The effects of lipid-lowering drug therapy on cardiovascular responsiveness in type 2 diabetic patients. AB - Type 2 diabetes is associated with a high prevalence of dyslipidaemia and a high incidence of cardiovascular disease. Lipid lowering therapy with HMG Co-A reductase inhibitors (statins) reduce the risk of cardiovascular events in type 2 diabetic and non-diabetic patients, effects which are believed to be partly due to improvements in vascular function. The aetiology of abnormal vascular function in type 2 diabetics is likely to be multifactorial and the pattern of vascular dysfunction in type 2 diabetes may differ from that which occurs in non-diabetic patients with dyslipidaemia. Abnormalities in endothelium derived hyperpolarising factor (EDHF) mediated vasodilation in resistance vessels may be more prominent in both type 1 and type 2 diabetes than in non-diabetic patients with endothelial dysfunction. The effects of lipid lowering therapy on vascular responsiveness may differ in type 2 diabetic patients from those found in non-diabetic patients. Statin therapy does not appear to improve responses to endothelial dependent vasodilators in type 2 diabetics, but may alter the ratio between nitric oxide (NO) and EDHF mediated responses. Fibrate therapy improves flow mediated dilation of brachial arteries in type 2 diabetic patients, but only appears to improve endothelium dependant vasodilator responses in resistance vessels when given in conjunction with co-enzyme Q. PMID- 16367878 TI - Advances in pancreatic islet transplantation in humans. AB - With recent advances in methods of islet isolation and the introduction of more potent and less diabetogenic immunosuppressive therapies, islet transplantation has progressed from research to clinical reality. Presently, several international centres have demonstrated successful clinical outcomes with high rates of insulin independence after islet transplantation. Ongoing refinements in donor pancreas procurement and processing, developments in islet isolation and purification technology, and advances in novel immunological conditioning and induction therapies have led to the acceptance of islet transplantation as a safe and effective therapy for patients with type 1 diabetes. This review provides a historical perspective of islet transplantation, outlines the recent advances and current clinical outcomes, and addresses the present challenges and future directions in clinical islet transplantation. PMID- 16367879 TI - Insulin detemir: a new basal insulin analogue. AB - Basal insulin therapy is an integral part of the intensive management of type 1 diabetes and it is also often used in type 2 diabetes. An ideal insulin regimen in patients with diabetes would mirror the 24-h insulin profile of a non-diabetic person, thereby preventing hyperglycaemia without inducing hypoglycaemia. Until recently, available insulins have pharmacokinetic disadvantages, compared to physiological insulin secretion. Insulin detemir is a new basal insulin analogue recently available for commercial use in the UK. Clinical trials have demonstrated lower fasting plasma glucose levels, lower variability in plasma glucose, predictable action profile and a reduced risk of nocturnal hypoglycaemia and weight gain, compared to conventional basal insulins. This study reviews the properties and potential use of insulin detemir. PMID- 16367880 TI - Pioglitazone and reductions in post-challenge glucose levels in patients with type 2 diabetes. AB - AIM: Pioglitazone (PIO) has been shown to decrease insulin resistance in patients with type 2 diabetes, resulting in lowered blood glucose concentrations, lowered plasma insulin levels and lowered haemoglobin A1C (A1C) values. Postprandial glucose control has been recently recognized as an important target for reducing overall glycemic burden in patients with type 2 diabetes. Some authors assert that reductions in postprandial glucose levels may lead to a decrease in cardiovascular risk, one of the major complications associated with diabetes. METHODS: Data were analysed from a 26-week PIO monotherapy study of 88 patients who underwent a 3-h oral glucose tolerance test (75 g dose) at baseline and last measurement. Change from baseline in area under the curve (AUC) values and hourly glucose concentrations were calculated and analysed at both time points for four medication groups: placebo group and PIO 15, 30 and 45 mg groups. Changes from baseline in fasting plasma glucose (FPG) and A1C also were reported. RESULTS: Glucose AUC was significantly (p < 0.05) different from baseline at 15, 30 and 45 mg doses of PIO. In addition, when compared to placebo, PIO (15, 30 and 45 mg) significantly decreased post-challenge blood glucose AUC (p < 0.05). The mean hourly blood glucose levels at last measurement for PIO 15, 30 and 45 mg all were significantly lower (p < 0.05) than placebo at all four time points. In addition, PIO significantly (p < 0.05) reduced FPG and A1C from baseline to last measurement in a dose-related fashion. CONCLUSIONS: PIO significantly reduced post-challenge glucose levels following an oral glucose challenge, leading to improvements in overall glycemic control. Postprandial glucose lowering is one of several metabolic effects of PIO in addition to decreasing insulin resistance and improving some lipids components. Whether these combined metabolic effects can lead to cardiovascular risk reductions may be confirmed by the pending results of cardiovascular outcomes studies with PIO. PMID- 16367881 TI - Biphasic insulin aspart 30 plus metformin: an effective combination in type 2 diabetes. AB - AIM: This study compared glycaemic control achieved with biphasic insulin aspart 30 (BIAsp 30) monotherapy, BIAsp 30 plus metformin and glibenclamide plus metformin in patients with type 2 diabetes not adequately controlled with metformin. METHODS: In this multinational, open-labelled, parallel group, 16-week trial, 341 patients (patients not adequately controlled with metformin for at least 1 month) with type 2 diabetes were studied. Patients were randomized to receive BIAsp 30, twice daily (n = 107 exposed to treatment), or BIAsp 30, twice daily, plus metformin (n = 108) or glibenclamide plus metformin (n = 114). The primary endpoint was HbA(1c) at end of trial; adverse events, hypoglycaemia episodes, blood lipids and weight were also monitored. RESULTS: In the total population (HbA(1c) 7.5-13.0% at screening), end-of-trial HbA(1c) levels were lower in patients receiving BIAsp 30 plus metformin compared with those receiving BIAsp 30 only [mean treatment difference (+/-s.e.m), 0.39 +/- 0.15%, p = 0.007]. In a subpopulation (HbA(1c) > or = 9.0% at baseline, n = 193), patients receiving BIAsp 30 plus metformin had significantly lower HbA(1c) levels at the end of the trial compared with those receiving glibenclamide plus metformin (treatment difference, 0.46 +/- 0.21%, p = 0.027). Mean body weight (+/-s.d) at the end of the trial was significantly lower in patients receiving glibenclamide plus metformin compared with those receiving BIAsp 30 only (84.3 +/- 13.3 kg vs. 88.9 +/- 16.9 kg, p < 0.001). No major hypoglycaemic episodes were recorded during the trial, and incidence rates for minor and symptoms-only hypoglycaemia were low and similar between treatment groups (0.03-0.04 events/patient/week). CONCLUSION: BIAsp 30 added to metformin could be an appropriate therapeutic option for achieving good glycaemic control, compared with the addition of a second oral agent, particularly where HbA(1c) > or = 9%. PMID- 16367882 TI - Effect of early addition of rosiglitazone to sulphonylurea therapy in older type 2 diabetes patients (>60 years): the Rosiglitazone Early vs. SULphonylurea Titration (RESULT) study. AB - AIM: To compare the efficacy, safety and tolerability of adding rosiglitazone (RSG) vs. sulphonylurea (SU) dose escalation in older type 2 diabetes mellitus (T2DM) patients inadequately controlled on SU therapy. METHODS: A total of 227 T2DM patients from 48 centres in the USA and Canada, aged > or =60 years, were randomized to receive RSG (4 mg) or placebo once daily in combination with glipizide 10 mg twice daily for 2 years in a double-blind, parallel-group study. Previous SU monotherapy was (1/4) to (1/2) maximum recommended dose for > or =2 months prior to screening with fasting plasma glucose (FPG) > or =7.0 and < or =13.9 mmol/l. Treatment options were individualized, and escalation of study medication was specifically defined. RESULTS: Disease progression (time to reach confirmed FPG > or =10 mmol/l while on maximum doses of both glipizide and study medication or placebo) was reported in 28.7% of patients uptitrating SU plus placebo compared with only 2.0% taking RSG and SU combination (p < 0.0001). RSG + SU significantly decreased HbA(1c), FPG, insulin resistance, plasma free fatty acids and medical care utilization and improved treatment satisfaction compared with uptitrated SU. CONCLUSIONS: Addition of RSG to SU in older T2DM patients significantly improved glycaemic control and reduced disease progression compared with uptitrated SU alone but without increasing hypoglycaemia. These benefits were associated with increased patient treatment satisfaction and reduced medical care utilization with regards to emergency room visits and length of hospitalization. Early addition of RSG is an effective treatment option for older T2DM patients inadequately controlled on submaximal SU monotherapy. PMID- 16367883 TI - Attainment of glycaemic goals in type 2 diabetes with once-, twice-, or thrice daily dosing with biphasic insulin aspart 70/30 (The 1-2-3 study). AB - AIM: This observational study in patients with type 2 diabetes failing oral agent therapy with or without basal insulin was conducted to assess whether addition and self-titration of biphasic insulin aspart 70/30 (BIAsp 30) could achieve American Association of Clinical Endocrinologists (AACE)/International Diabetes Federation (IDF) and American Diabetes Association (ADA) glycemic targets (HbA(1c)< or =6.5 and <7%). METHODS: Enrolled patients (n = 100, HbA(1c)> or =7.5 and < or =10%) were > or =18 years of age, had diabetes > or =12 months and had received a stable antidiabetic regimen for at least 3 months [minimum of two oral antidiabetic drugs (OADs) or at least one OAD plus once-daily basal insulin < or =60 U]. Patients discontinued prior basal insulin and added one injection of BIAsp 30 (12 U or 70-100% of prior basal insulin dose within 15 min of dinner initiation). Patients self-titrated their BIAsp 30 dose with investigator guidance every 3 or 4 days to achieve pre-breakfast fasting blood glucose (FBG) of 80-110 mg/dl. At 16 weeks, a pre-breakfast injection of 6 U of BIAsp 30 was added if week 15 HbA(1c) exceeded 6.5%; the added dose was titrated to achieve pre-dinner BG of 80-110 mg/dl. After an additional 16 weeks, 3 U of pre-lunch BIAsp 30 was added if HbA(1c) exceeded 6.5%. This added dose was adjusted based on 2-h post-lunch BG to achieve postprandial glucose of 100-140 mg/dl. Subjects achieving an HbA(1c)< or =6.5% at 15 and 31 weeks completed the study at weeks 16 and 32 respectively. RESULTS: Addition of once-daily BIAsp 30 before dinner enabled 21% of the patients to achieve AACE and IDF targets (HbA(1c)< or =6.5%) and 41% to achieve ADA targets (HbA(1c) <7%). With two daily injections of BIAsp 30, these glycaemic goals were achieved by 52 and 70% of subjects. With three daily BIAsp 30 injections, 60% of patients achieved HbA(1c)< or =6.5%, and 77% achieved HbA(1c) <7.0%. CONCLUSIONS: This clinical trial demonstrates that initiation of once-daily BIAsp 30 to type 2 diabetes patients poorly controlled on various OAD regimens was an effective treatment approach for achieving glycaemic goals. Additional patients safely achieved these goals by increasing the number of BIAsp 30 injections from one to two, and then, if uncontrolled, from two to three doses per day. Eventually, most patients previously uncontrolled on OADs with or without basal insulin were controlled by the addition and vigorous titration of BIAsp 30 to oral agent therapy. PMID- 16367884 TI - Insulin status differentially affects energy transduction in cardiac mitochondria from male and female rats. AB - AIM: The incidence of coronary heart diseases (CHD), congestive heart failure (CHF) and myocardial infarction is higher in diabetic patients than in non diabetic groups, with these incidences being more in women than in the men. Hence, we examined involvement of mitochondrial energy transduction functions. METHODS: Mitochondrial energy metabolism in cardiomyopathy was studied using streptozotocin (STZ)-diabetic male and female rats as the model system. Effects of insulin treatment were also evaluated. RESULTS: The body and heart weights decreased in both male and female diabetic rats. Insulin treatments resulted in significant increase in the body and heart weights in the female rats. Mitochondrial respiration rates with all the substrates tested decreased in diabetic condition in both males and females. Treatment with two dose-regimens of insulin had differential restorative effect on mitochondrial substrate oxidation in the males but caused hyper-stimulation in the females. Diabetic state brought about 19% decrease in the cytochrome aa(3) content in the female rats. Treatment with 0.6 units of insulin significantly increased the cytochrome contents in general in both the sexes whereas higher dose (1.0 unit) caused decrease in the cytochromes content in the females. Diabetic state resulted in decreased dehydrogenases activities; insulin treatments had differential effect on the dehydrogenase activity in the males and the females. The results suggest that insulin treatment-induced hyper-stimulation of respiration in female rats may lead to increased production of reactive oxygen species. Besides, increased formation of advanced glycosylated end products may further lead to increased risk of CHF and CHD. CONCLUSIONS: The results suggest that differential effects of STZ-diabetes and insulin treatments in the female rats than in males may be the underlying cause for increased incidence of diabetic cardiomyopathies in the females. PMID- 16367885 TI - IRS1, KCNJ11, PPARgamma2 and HNF-1alpha: do amino acid polymorphisms in these candidate genes support a shared aetiology between type 1 and type 2 diabetes? AB - AIMS: Type 1 diabetes mellitus (T1DM) is a chronic disorder primarily triggered by environmental and immunological factors in genetically susceptible individuals. Despite the fact that there are indications of common aetiological features of T1DM and type 2 diabetes (T2DM), variation in genes involved in insulin secretion and insulin signalling has to a large extent been ignored as potential modifiers in the pathogenesis of T1DM. Recent studies suggest, however, that proven T2DM susceptibility gene variants may be involved in the pathogenesis of T1DM. The objective of this study was to estimate the impact of four selected amino acid polymorphisms -IRS-1 Gly972Arg, Kir6.2 Glu23Lys, HNF-1alpha Ala98Val and PPARgamma2 Pro12Ala in a Danish population of T1DM families. METHODS: All variants were genotyped in 490 simplex- and multiplex-T1DM families applying polymerase chain reaction-restriction fragment length polymorphism, and results were evaluated by means of a transmission disequilibrium test (TDT) analysis. RESULTS: TDT analysis revealed that the Arg972 IRS-1, the Lys23 Kir6.2 and the Val98 HNF-1alpha variants were transmitted from heterozygous parents to affected probands at frequencies of 49.1%, 47.0% and 54.1%, respectively (p > 0.05 for all). This was similar to the rate of transmission to unaffected siblings. The transmission rate of the Ala12 PPARgamma2 variant to affected probands was 46.5% (p > 0.05) which differed significantly from the transmission to unaffected offspring (p = 0.024). A combined analysis of the present and published pertinent data of 1691 transmissions showed a significantly decreased transmission of the PPARgamma2 Ala12 allele to affected probands (p = 0.0045). CONCLUSIONS: The Pro12Ala variant of PPARgamma2 is associated with T1DM, the minor Ala allele conferring a reduced risk. This same finding has been reported in patients with T2DM. PMID- 16367886 TI - Differentiation-dependent regulation of the cyclooxygenase cascade during adipogenesis suggests a complex role for prostaglandins. AB - AIM: A thorough understanding of the mechanisms of adipocyte differentiation and metabolism is important for the prevention and/or treatment of obesity and its complications, including type 2 diabetes mellitus. A complex role for prostaglandins (PGs) in adipogenesis is suggested. We examined the expression and cellular localization of enzymes in the cyclooxygenase (COX) cascade that synthesize PGs as well as the PG profile as a function of differentiation status in 3T3-L1 cells. METHODS: Murine 3T3-L1 preadipocytes were used as a model for studies of adipocyte differentiation induced by a hormone cocktail and compared with the parental fibroblastic line NIH 3T3. Both cell lines were incubated in maintenance medium or differentiation medium. Nine days after differentiation, the expression of enzymes in the COX cascade was evaluated by immunoblot analysis, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry, and PG formation was examined using enzyme immunoassay. RESULTS: A differentiation-dependent diminution of COX-1 and COX-2 mRNA and cognate proteins in 3T3-L1 cells was observed. PG release, including PGE(2), 6 keto PGF(1alpha), PGD(2) and 15d-PGJ(2), significantly decreased following differentiation in 3T3-L1 cells (anova/Tukey, p < 0.05). However, microsomal PGE synthase (mPGES) and lipocalin-type PGD synthase (L-PGDS) were selectively upregulated. Immunocytochemistry revealed that COX-1 and COX-2 became intracellularly more diffuse upon differentiation, whereas mPGES was redistributed to the nuclear compartment. CONCLUSIONS: Regulation of PG formation and COX-2 expression in 3T3-L1 cells is differentiation-dependent and involves changes in the levels of gene expression of the individual isoforms as well as redistribution of the enzymes within cellular compartments. PMID- 16367887 TI - Prediction of cardiovascular and total mortality in Chinese type 2 diabetic patients by the WHO definition for the metabolic syndrome. AB - AIM: The aim of this study is to investigate the prevalence of metabolic syndrome (MES) in type 2 diabetic patients and the predictive values of the World Health Organization (WHO) and National Cholesterol Education Programme (NCEP) definitions and the individual components of the MES on total and cardiovascular mortality. METHODS: A prospective analysis of a consecutive cohort of 5202 Chinese type 2 diabetic patients recruited between July 1994 and April 2001. RESULTS: The prevalence of the MES was 49.2-58.1% depending on the use of various criteria. There were 189 deaths (men: 100 and women: 89) in these 5205 patients during a median (interquartile range) follow-up period of 2.1 (0.3-3.6 years). Of these, 164 (87%) were classified as cardiovascular deaths. Using the NCEP criterion, patients with MES had a death rate similar to those without (3.51 vs. 3.85%). By contrast, based on the WHO criteria, patients with MES had a higher mortality rate than those without (4.3 vs. 2.4%, p = 0.002). Compared to patients with neither NCEP- nor WHO-defined MES, only the group with MES defined by the WHO, but not NCEP, criterion had significantly higher mortality rate (2.6 vs. 6.8%, p < 0.001). Using Cox regression analysis, only age, duration of diabetes and smoking were identified as independent factors for cardiovascular or total death. Among the various components of MES, hypertension, low BMI and albuminuria were the key predictors for these adverse events. CONCLUSIONS: In Chinese type 2 diabetic patients, the WHO criterion has a better discriminative power over the NCEP criterion for predicting death. Among the various components of the MES defined either by WHO or NCEP, hypertension, albuminuria and low BMI were the main predictors of cardiovascular and total mortality. PMID- 16367888 TI - Effects of metformin and oleic acid on adipocyte expression of resistin. AB - AIM: The adipocyte-secreted hormone resistin has been implicated in obesity induced insulin resistance and type 2 diabetes, but pharmacological and dietary factors that regulate resistin gene expression and the effects of resistin on cellular glucose uptake in muscle have not been clearly defined. METHODS: Expression of resistin mRNA was studied in differentiated 3T3-L1 adipocytes by using real-time semiquantitative reverse transcription-polymerase chain reaction. The effects of resistin on insulin-stimulated and insulin-independent 2 deoxyglucose uptake were evaluated in L6 muscle cells. RESULTS: Insulin 1 microm and rosiglitazone 10 microm markedly reduced resistin mRNA expression (relative to the control gene TF2D) by 4.7-fold (p < 0.05) and 5.3-fold (p < 0.02), respectively. Similar reductions in resistin mRNA were demonstrated with metformin 100 microm (6.2-fold reduction, p < 0.02) and oleic acid 100 microm (3.9-fold reduction, p < 0.03). Resistin 1 microm significantly reduced maximum insulin-stimulated 2-deoxyglucose uptake in L6 cells from 634 to 383% (relative to 100% for control, p < 0.001), and co-administration of rosiglitazone had no effect on resistin-induced insulin resistance. In the absence of insulin, however, resistin increased glucose uptake dose-dependently (e.g., 1.75-fold at 5 microm, p < 0.001) via a mitogen-activated protein kinase-dependent pathway. CONCLUSIONS: These results demonstrate that various glucose-lowering therapies and oleic acid reduce resistin gene expression in isolated adipocytes, and that resistin impairs insulin-stimulated glucose uptake in skeletal muscle-derived cells. PMID- 16367889 TI - Long-term glycaemic efficacy and weight changes associated with thiazolidinediones when added at an advanced stage of type 2 diabetes. AB - OBJECTIVE: To describe the outcome of 35 patients with type 2 diabetes prospectively followed for 6 years after the addition of a thiazolidinedione (TZD) to a failing regimen of a sulphonylurea and metformin -- triple oral therapy. METHODS: Study patients were assessed for the need for the addition of insulin to their regimen, and follow-up clinical and laboratory findings were analysed. RESULTS: At a mean follow-up of 72 +/- 1.5 months (range 53-80), 18 (51%) of patients remained well controlled on triple oral therapy with a mean glycosylated haemoglobin (HbA1c) value of 6.9 +/- 0.2% (upper limit of normal 6.2%). In 17 other patients, triple oral therapy failed and the use of insulin was necessary after a mean duration of 38 (range 18-68) months. The mean HbA1c in these patients was 8.0 +/- 0.3%. The group that was maintained on triple oral therapy gained 15.2 +/- 1.9 lbs over the 6-year study which was significantly higher than the baseline weight. Alternatively, the group that failed and had insulin added to their therapy gained 20.2 +/- 4.5 lbs over the same period which was also significantly different from baseline but not from the triple oral therapy group. Although after 3 years a trend towards weight loss occurred in the triple oral therapy group, the insulin-added group continued to gain weight. Stimulated C-peptide levels increased significantly in the triple therapy group from 3.6 +/- 0.9 to 4.3 +/- 1.2 ng/ml and had not increased or decreased non significantly from 3.7 +/- 0.8 to 3.2 +/- 0.6 ng/ml at the time of insulin initiation in the insulin-requiring group. CONCLUSION: When used late in the course of type 2 diabetes, TZDs result in improved and prolonged glycaemic control which persisted for a median time of 6 years. Weight gain with TZDs peaks and then plateaus (and even trends downwards) at 3 years, although the addition of insulin to a failing oral therapy regimen results in a further and continuing weight gain in spite of inferior glycaemic control. Continuing glycaemic control with triple oral therapy is dependent on preservation or augmentation of endogenous insulin production. PMID- 16367891 TI - Depression in Parkinson's disease -- a review. AB - Major depression is present, at any given time, in 20-40% of Parkinson's disease (PD) patients, several times the prevalence in the general population. In addition, depression may precede the diagnosis of PD. These observations and reports of depression during deep brain stimulation of regions contiguous to the substantia nigra, as well as reports of dopamine agonist improving depression, suggest depression, rather than being mainly a psychological reaction to a debilitating disease, is part of PD. It is postulated that mesolimbic and mesocortical dopaminergic pathways that mediate affect, behavior, and cognition, contribute to depression in PD. PMID- 16367892 TI - Parkin gene variations in late-onset Parkinson's disease: comparison between Norwegian and German cohorts. AB - OBJECTIVES: Mutations in the Parkin gene can cause autosomal recessive early onset Parkinson's disease (PD). Recently, Parkin mutations were also suggested to play a role in the commoner late-onset forms of PD. METHODS: We compared a German cohort of PD patients (95) with a Norwegian cohort of PD patients (96). Both cohorts have predominant late-onset form of PD. Mutation and polymorphism frequencies were compared via single-strand conformation polymorphism and sequence analyses. RESULTS: Three heterozygous missense mutations (Arg256Cys, Arg402Cys and Thr240Met) were found in late-onset PD patients in the German patient cohort (1.6%). A missense mutation (Arg402Cys) was also found in one of 149 healthy control subjects (0.3%). Only one heterozygous missense mutation (Arg256Cys) was identified in a Norwegian patient suffering from late-onset PD (0.5%). The frequencies of four known single nucleotide polymorphisms significantly differ between the two distant European populations. CONCLUSION: The results support the hypothesis that heterozygous mutations in the Parkin gene may act as susceptibility alleles for late-onset forms of PD in rare cases. PMID- 16367893 TI - Apolipoprotein epsilon4 advances appearance of psychosis in patients with Parkinson's disease. AB - BACKGROUND: Psychosis is one of the most serious complications of advanced parkinsonism, but many patients are spared. The genetic factors predisposing to psychosis are unclear. OBJECTIVES: To assess the association between apolipoprotein E (APOE) polymorphism and the development of psychosis in patients with Parkinson's disease (PD). METHODS: Eighty-seven patients with advanced PD were assessed. Psychosis was diagnosed in 50 patients who manifested paranoid delusions, hallucinations without insight, or disorders of perception. Time of onset of psychosis was retrieved from the medical records and caregivers' recall. APOE genotype was determined by restriction enzyme digests of amplified alleles. Cox models of logistic regression and Kaplan-Meier survival curves were used to assess factors determining early development of psychosis. RESULTS: APOE epsilon3/epsilon4 allele was carried by 20 patients (14 with psychosis), epsilon2/epsilon3 by 11 patients (10 with psychosis), epsilon3/epsilon3 by 55 patients (25 with psychosis) and epsilon2/epsilon4 by one patient who had psychosis. The mean age of onset of PD symptoms was 60.0 +/- 12.5 years. The mean duration of motor symptoms at the onset of psychosis was 7.3 +/- 4.3 years for the 15 patients harboring an APOE epsilon4 allele and 10.1 +/- 6.2 years among those who did not carry APOE epsilon4 (n = 35). The APOE epsilon4 allele was significantly associated with earlier onset of psychosis (P < 0.05) when the age of onset of motor symptoms and presence of dementia were included in the Cox regression model. Carrying the APOE epsilon4 allele was a significant risk factor for earlier appearance of psychosis with a hazard ratio of 3.24 (95% CI 1.62 6.46) while dementia by itself did not increase the risk. CONCLUSION: Parkinson's disease patients who carry the APOE epsilon4 allele develop psychosis earlier. PMID- 16367894 TI - Efficacy and safety of high-dose cabergoline in Parkinson's disease. AB - OBJECTIVES: To assess the efficacy and safety of high-dose (up to 20 mg/day) cabergoline in Parkinson's disease (PD) patients with motor fluctuations and/or dyskinesias. MATERIALS AND METHODS: Thirty-four PD patients had cabergoline up titrated and their levodopa (L-dopa) reduced over a maximum of 20 weeks, followed by at least 6 weeks steady cabergoline dosing. Primary endpoint was change in mean hyperkinesia intensity at the final visit (week 26). RESULTS: Mean (+/- SD) cabergoline was increased from 6.43 +/- 2.66 to 12.78 +/- 5.67 mg/day and mean L dopa reduced from 606.6 +/- 263.9 to 370.6 +/- 192.5 mg/day. A significant reduction (P < 0.001) in mean hyperkinesia intensity occurred from baseline (day 0) to week 26. Improvements in 'on with dyskinesias', mean dystonia intensity (P < 0.05), time spent in 'severe off' condition, severity of 'off' periods as well as clinical/patient global impression, and health-related quality of life were observed. Twenty-four drug-related adverse events were recorded of which four were regarded as serious. CONCLUSION: High-dose cabergoline was well tolerated and provided significant improvements in the Parkinson symptomatology and a reduced requirement for L-dopa. PMID- 16367895 TI - The Swedish Malignant Middle cerebral artery Infarction Study: long-term results from a prospective study of hemicraniectomy combined with standardized neurointensive care. AB - OBJECTIVES: Hemicraniectomy in patients with malignant middle cerebral artery (mMCA) infarct may be life-saving. The long-term prognosis is unknown. METHODS: Patients with mMCA infarct treated with hemicraniectomy between 1998 and 2002 at three hospitals were included. The criterion for surgical intervention was if the patients deteriorated from awake to being responding to painful stimuli only. All patients were followed for at least 1 year. Outcome was defined as alive/dead, walkers/non-walkers or modified Rankin Scale (mRS) score T is not involved in either the susceptibility to Chlamydia trachomatis infection or the development of tubal pathology. AB - BACKGROUND: The functional polymorphism -260 C>T in the LPS sensing TLR4 co receptor CD14 gene enhances the transcriptional activity and results in a higher CD14 receptor density. Individuals carrying the T/T genotype also have significantly higher serum levels of soluble CD14. The T allele of this polymorphism has recently been linked to Chlamydia pneumoniae infection. We investigated the role of the CD14 -260 C>T polymorphism in the susceptibility to and severity (defined as subfertility and/or tubal pathology) of C. trachomatis infection in Dutch Caucasian women. METHODS: The different CD14 -260 C>T genotypes were assessed by PCR-based RFLP analysis in three cohorts: 1) A cohort (n = 576) of women attending a STD clinic, 2) a cohort (n = 253) of women with subfertility, and 3) an ethnically matched control cohort (n = 170). The following variables were used in the analysis: In cohort 1 the CT-DNA status, CT IgG serology status, self-reported symptoms and in cohort 2, the CT IgG serology status and the tubal status at laparoscopy. RESULTS: In the control cohort the CC, CT and TT genotype distribution was: 28.2%, 48.2%, and 23.5% respectively. No differences were found in the overall prevalence of CD14 -260 genotypes (28.1%, 50.7%, and 21.2%) in cohort 1 when compared to the control cohort. Also no differences were observed in women with or without CT-DNA, with or without serological CT responses, with or without symptoms, or in combinations of these three variables. In subfertile women with tubal pathology (cohort 2, n = 50) the genotype distribution was 28.0%, 48.0%, and 24.0% and in subfertile women without tubal pathology (n = 203), 27.6%, 49.3% and 23.2%. The genotype distribution was unchanged when CT IgG status was introduced in the analyses. CONCLUSION: The CD14 -260 C>T genotype distributions were identical in all three cohorts, showing that this polymorphism is not involved in the susceptibility to or severity of sequelae of C. trachomatis infection. PMID- 16368003 TI - Involvement of claudin-7 in HIV infection of CD4(-) cells. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection of CD4(-) cells has been demonstrated, and this may be an important mechanism for HIV transmission. RESULTS: We demonstrated that a membrane protein, claudin-7 (CLDN-7), is involved in HIV infection of CD4(-) cells. A significant increase in HIV susceptibility (2 to 100-fold) was demonstrated when CLDN-7 was transfected into a CD4(-) cell line, 293T. In addition, antibodies against CLDN-7 significantly decreased HIV infection of CD4(-) cells. Furthermore, HIV virions expressing CLDN-7 on their envelopes had a much higher infectivity for 293T CD4(-) cells than the parental HIV with no CLDN-7. RT-PCR results demonstrated that CLDN-7 is expressed in both macrophages and stimulated peripheral blood leukocytes, suggesting that most HIV virions generated in infected individuals have CLDN-7 on their envelopes. We also found that CLDN-7 is highly expressed in urogenital and gastrointestinal tissues. CONCLUSION: Together these results suggest that CLDN-7 may play an important role in HIV infection of CD4(-) cells. PMID- 16368004 TI - Multiple major increases and decreases in mitochondrial substitution rates in the plant family Geraniaceae. AB - BACKGROUND: Rates of synonymous nucleotide substitutions are, in general, exceptionally low in plant mitochondrial genomes, several times lower than in chloroplast genomes, 10-20 times lower than in plant nuclear genomes, and 50-100 times lower than in many animal mitochondrial genomes. Several cases of moderate variation in mitochondrial substitution rates have been reported in plants, but these mostly involve correlated changes in chloroplast and/or nuclear substitution rates and are therefore thought to reflect whole-organism forces rather than ones impinging directly on the mitochondrial mutation rate. Only a single case of extensive, mitochondrial-specific rate changes has been described, in the angiosperm genus Plantago. RESULTS: We explored a second potential case of highly accelerated mitochondrial sequence evolution in plants. This case was first suggested by relatively poor hybridization of mitochondrial gene probes to DNA of Pelargonium hortorum (the common geranium). We found that all eight mitochondrial genes sequenced from P. hortorum are exceptionally divergent, whereas chloroplast and nuclear divergence is unexceptional in P. hortorum. Two mitochondrial genes were sequenced from a broad range of taxa of variable relatedness to P. hortorum, and absolute rates of mitochondrial synonymous substitutions were calculated on each branch of a phylogenetic tree of these taxa. We infer one major, approximately 10-fold increase in the mitochondrial synonymous substitution rate at the base of the Pelargonium family Geraniaceae, and a subsequent approximately 10-fold rate increase early in the evolution of Pelargonium. We also infer several moderate to major rate decreases following these initial rate increases, such that the mitochondrial substitution rate has returned to normally low levels in many members of the Geraniaceae. Finally, we find unusually little RNA editing of Geraniaceae mitochondrial genes, suggesting high levels of retroprocessing in their history. CONCLUSION: The existence of major, mitochondrial-specific changes in rates of synonymous substitutions in the Geraniaceae implies major and reversible underlying changes in the mitochondrial mutation rate in this family. Together with the recent report of a similar pattern of rate heterogeneity in Plantago, these findings indicate that the mitochondrial mutation rate is a more plastic character in plants than previously realized. Many molecular factors could be responsible for these dramatic changes in the mitochondrial mutation rate, including nuclear gene mutations affecting the fidelity and efficacy of mitochondrial DNA replication and/or repair and- consistent with the lack of RNA editing--exceptionally high levels of "mutagenic" retroprocessing. That the mitochondrial mutation rate has returned to normally low levels in many Geraniaceae raises the possibility that, akin to the ephemerality of mutator strains in bacteria, selection favors a low mutation rate in plant mitochondria. PMID- 16368005 TI - Comparison of foot orthoses made by podiatrists, pedorthists and orthotists regarding plantar pressure reduction in The Netherlands. AB - BACKGROUND: There is a need for evidence of clinical effectiveness of foot orthosis therapy. This study evaluated the effect of foot orthoses made by ten podiatrists, ten pedorthists and eleven orthotists on plantar pressure and walking convenience for three patients with metatarsalgia. Aims were to assess differences and variability between and within the disciplines. The relationship between the importance of pressure reduction and the effect on peak pressure was also evaluated. METHODS: Each therapist examined all three patients and was asked to rate the 'importance of pressure reduction' through a visual analogue scale. The orthoses were evaluated twice in two sessions while the patient walked on a treadmill. Plantar pressures were recorded with an in-sole measuring system. Patients scored walking convenience per orthosis. The effects of the orthoses on peak pressure reduction were calculated for the whole plantar surface of the forefoot and six regions: big toe and metatarsal one to five. RESULTS: Within each discipline there was an extensive variation in construction of the orthoses and achieved peak pressure reductions. Pedorthists and orthotists achieved greater maximal peak pressure reductions calculated over the whole forefoot than podiatrists: 960, 1020 and 750 kPa, respectively (p < .001). This was also true for the effect in the regions with the highest baseline peak pressures and walking convenience rated by patients A and B. There was a weak relationship between the 'importance of pressure reduction' and the achieved pressure reduction for orthotists, but no relationship for podiatrists and pedorthotists. CONCLUSION: The large variation for various aspects of foot orthoses therapy raises questions about a consistent use of concepts for pressures management within the professional groups. PMID- 16368006 TI - Effects among healthy subjects of the duration of regularly practicing a guided imagery program. AB - BACKGROUND: We examined a large number of healthy adults in the general community who had individually participated in a guided imagery (GI) program daily and for various durations, to examine the psychophysiological effects of a GI program within a healthy group. METHODS: We studied 176 subjects who had participated in sessions that were part of a guided imagery program, and who had practiced GI at home for 20 minutes once daily in a quiet place after mastering GI in the group sessions. The average duration of GI practiced at home was 6.88 +/- 14.06 months (n = 138, range: 0 to 72). The Multiple Mood Scale (MMS), Betts (1909) Shortened Questionnaire on Mental Imagery (QMI), and a visual analog scale (VAS) of imagery vividness, salivary cortisol (CS) levels, general stress and general health were used in the sessions. RESULTS: We examined the relationship between the duration of daily GI practiced at home and MMS, QMI, CS, general health, and general stress at baseline. The subjects who had practiced GI at home longer had lower negative mood scores at baseline and lower severity of stress, and higher positive mood at baseline (both at a session and at home), general health, and QMI scores at baseline. The MMS change during a session and the duration of daily GI practiced at home were not correlated. Repeated-measures analysis of covariance showed that the duration of daily GI practiced as the covariate was not associated with changes in the three CS levels. CONCLUSION: Although regularly practicing a GI program daily for 20 min did not affect the CS level or mood during a GI session for several hours, it kept a good condition of the general mental, physical well-being and their overall stress of the practitioners as they had practiced it for long duration. We postulate that subjects who have the high ability of imaging vividness showed the better mood, health status and less stress than those subjects who have the low ability of it did. The ability of image vividness of the long-term regular practitioners of GI was higher than its short-term or inexperienced practitioners, which allowed practitioners to produce more comfortable imagery. Consequently, the longer the duration that they had practiced GI program once a day regularly, the lower scores of their stress were and the higher scores of their health were. We suggest that the regular daily practice of a GI program might be connected to less stress and better health. PMID- 16368007 TI - Scutellaria baicalensis decreases ritonavir-induced nausea. AB - BACKGROUND: Protease inhibitors, particularly ritonavir, causes significant gastrointestinal disturbances such as nausea, even at low doses. This ritonavir induced nausea could be related to its oxidative stress in the gut. Alleviation of drug-induced nausea is important in effectively increasing patients' compliance and improving their quality of life. Conventional anti-emetic drugs can only partially abate the symptoms in these patients, and their cost has also been a concern. Rats respond to nausea-producing emetic stimuli by increasing consumption of non-nutritive substances like kaolin or clay, a phenomenon known as pica. In this study, we used this rat pica model to evaluate the effects of Scutellaria baicalensis, a commonly used oriental herbal medicine, on ritonavir induced nausea. RESULTS: Rats treated with 20 mg/kg ritonavir significant caused increases of kaolin consumption at 24 to 48 hr (P < 0.01). Pretreatment with 0.3 and 3 mg/kg Scutellaria baicalensis extract significantly decreased ritonavir induced kaolin intake in a dose-related manner (P < 0.01). Compared to vehicle treatment, the extract completely prevented ritonavir-induced kaolin consumption at dose 3 mg/kg. The area under the curves (AUC) for kaolin intake from time 0 to 120 hr for vehicle only, ritonavir only, SbE 0.3 mg/kg plus ritonavir, and SbE 3 mg/kg plus ritonavir were 27.3 g x hr, 146.7 g x hr, 123.2 g x hr, and 32.7 g x hr, respectively. The reduction in area under the curves of kaolin intake from time 0 to 120 hr between ritonavir only and SbE 0.3 mg/kg plus ritonavir, ritonavir only and SbE 3 mg/kg plus ritonavir were 16.0% and 77.7%, respectively. CONCLUSION: Scutellaria baicalensis significantly attenuated ritonavir-induced pica, and demonstrated a potential in treating ritonavir-induced nausea. PMID- 16368008 TI - Characterization of sub-nuclear changes in Caenorhabditis elegans embryos exposed to brief, intermediate and long-term anoxia to analyze anoxia-induced cell cycle arrest. AB - BACKGROUND: The soil nematode C. elegans survives oxygen-deprived conditions (anoxia; <.001 kPa O2) by entering into a state of suspended animation in which cell cycle progression reversibly arrests. The majority of blastomeres of embryos exposed to anoxia arrest at interphase, prophase and metaphase. The spindle checkpoint proteins SAN-1 and MDF-2 are required for embryos to survive 24 hours of anoxia. To further investigate the mechanism of cell-cycle arrest we examined and compared sub-nuclear changes such as chromatin localization pattern, post translational modification of histone H3, spindle microtubules, and localization of the spindle checkpoint protein SAN-1 with respect to various anoxia exposure time points. To ensure analysis of embryos exposed to anoxia and not post-anoxic recovery we fixed all embryos in an anoxia glove box chamber. RESULTS: Embryos exposed to brief periods to anoxia (30 minutes) contain prophase blastomeres with chromosomes in close proximity to the nuclear membrane, condensation of interphase chromatin and metaphase blastomeres with reduced spindle microtubules density. Embryos exposed to longer periods of anoxia (1-3 days) display several characteristics including interphase chromatin that is further condensed and in close proximity to the nuclear membrane, reduction in spindle structure perimeter and reduced localization of SAN-1 at the kinetochore. Additionally, we show that the spindle checkpoint protein SAN-1 is required for brief periods of anoxia induced cell cycle arrest, thus demonstrating that this gene product is vital for early anoxia responses. In this report we suggest that the events that occur as an immediate response to brief periods of anoxia directs cell cycle arrest. CONCLUSION: From our results we conclude that the sub-nuclear characteristics of embryos exposed to anoxia depends upon exposure time as assayed using brief (30 minutes), intermediate (6 or 12 hours) or long-term (24 or 72 hours) exposures. Analyzing these changes will lead to an understanding of the mechanisms required for initiation and maintenance of cell cycle arrest in respect to anoxia exposure time as well as order the events that occur to bring about anoxia-induced cell cycle arrest. PMID- 16368010 TI - Computer simulation of partitioning of ten pentapeptides Ace-WLXLL at the cyclohexane/water and phospholipid/water interfaces. AB - BACKGROUND: Peptide-membrane interactions play a key role in the binding, partitioning and folding of membrane proteins, the activity of antimicrobial and fusion peptides, and a number of other processes. To gain a better understanding of the thermodynamics of such interactions, White and Wimley created an interfacial hydrophobicity scale based of the transfer free energy from water to octanol or lipid bilayers of a series of synthetic peptapeptides (Ace-WLXLL, with X being any of the twenty natural amino acids) (White and Wimley (1996) Nat. Struct. Biol. 3, 842-848). In this study, we performed molecular dynamics simulations of a representative set of ten of these peptides (X = D, K, R, N, A, T, S, I, F and W) in two membrane mimetic interfaces: water-cyclohexane (10 ns) and a fully solvated dioleoylphosphatidylcholine (DOPC) bilayer (50 ns) using both constant pressure and constant area ensembles. We focus on partitioning of the ten peptides at the cyclohexane/water and lipid/water interfaces. RESULTS: The peptides rapidly equilibrate (< 2 ns) and partition at the cyclohexane/water interface. The X3 guest residue assumes average orientations that depend on the nature of the side chain. At the DOPC/water interface, dynamics is much slower and convergence is difficult to achieve on a 50 ns timescale. Nonetheless, all peptides partition to the lipid/water interface with distributions with widths of 1-2 nm. The peptides assume a broad range of side chain and backbone orientations and have only a small effect on the area of the unit cell. On average, hydrophobic guest residues partition deeper into the hydrophobic core than hydrophilic residues. In some cases the peptides penetrate sufficiently deep to somewhat affect the distribution of the C=C double bond in DOPC. The relative distribution of the X3 guest residue compared to W1 and L5 is similar in the water/cyclohexane and water/lipid simulations. Snapshots show mostly extended backbone conformations in both environments. There is little difference between simulations at a constant area of 0.66 nm2 and simulations at constant pressure that approximately yield the same average area of 0.66 nm2. CONCLUSION: These peptides were designed to assume extended conformations, which is confirmed by the simulations. The distribution of the X3 side chain depends on its nature, and can be determined from molecular dynamics simulations. The time scale of peptide motion at a phospholipids-water interface is too long to directly calculate the experimentally measured hydrophobicity scale to test and improve the simulation parameters. This should be possible at the water/cyclohexane interface and likely will become feasible in the future for the phospholipids/water case. PMID- 16368009 TI - An overlooked connection: serotonergic mediation of estrogen-related physiology and pathology. AB - BACKGROUND: In humans, serotonin has typically been investigated as a neurotransmitter. However, serotonin also functions as a hormone across animal phyla, including those lacking an organized central nervous system. This hormonal action allows serotonin to have physiological consequences in systems outside the central nervous system. Fluctuations in estrogen levels over the lifespan and during ovarian cycles cause predictable changes in serotonin systems in female mammals. DISCUSSION: We hypothesize that some of the physiological effects attributed to estrogen may be a consequence of estrogen-related changes in serotonin efficacy and receptor distribution. Here, we integrate data from endocrinology, molecular biology, neuroscience, and epidemiology to propose that serotonin may mediate the effects of estrogen. In the central nervous system, estrogen influences pain transmission, headache, dizziness, nausea, and depression, all of which are known to be a consequence of serotonergic signaling. Outside of the central nervous system, estrogen produces changes in bone density, vascular function, and immune cell self-recognition and activation that are consistent with serotonin's effects. For breast cancer risk, our hypothesis predicts heretofore unexplained observations of the opposing effects of obesity pre- and post-menopause and the increase following treatment with hormone replacement therapy using medroxyprogesterone. SUMMARY: Serotonergic mediation of estrogen has important clinical implications and warrants further evaluation. PMID- 16368011 TI - Comparison of gene expression signatures of diamide, H2O2 and menadione exposed Aspergillus nidulans cultures--linking genome-wide transcriptional changes to cellular physiology. AB - BACKGROUND: In addition to their cytotoxic nature, reactive oxygen species (ROS) are also signal molecules in diverse cellular processes in eukaryotic organisms. Linking genome-wide transcriptional changes to cellular physiology in oxidative stress-exposed Aspergillus nidulans cultures provides the opportunity to estimate the sizes of peroxide (O2(2-)), superoxide (O2*-) and glutathione/glutathione disulphide (GSH/GSSG) redox imbalance responses. RESULTS: Genome-wide transcriptional changes triggered by diamide, H2O2 and menadione in A. nidulans vegetative tissues were recorded using DNA microarrays containing 3533 unique PCR amplified probes. Evaluation of LOESS-normalized data indicated that 2499 gene probes were affected by at least one stress-inducing agent. The stress induced by diamide and H2O2 were pulse-like, with recovery after 1 h exposure time while no recovery was observed with menadione. The distribution of stress-responsive gene probes among major physiological functional categories was approximately the same for each agent. The gene group sizes solely responsive to changes in intracellular O2(2-), O2*- concentrations or to GSH/GSSG redox imbalance were estimated at 7.7, 32.6 and 13.0 %, respectively. Gene groups responsive to diamide, H2O2 and menadione treatments and gene groups influenced by GSH/GSSG, O2(2-) and O2*- were only partly overlapping with distinct enrichment profiles within functional categories. Changes in the GSH/GSSG redox state influenced expression of genes coding for PBS2 like MAPK kinase homologue, PSK2 kinase homologue, AtfA transcription factor, and many elements of ubiquitin tagging, cell division cycle regulators, translation machinery proteins, defense and stress proteins, transport proteins as well as many enzymes of the primary and secondary metabolisms. Meanwhile, a separate set of genes encoding transport proteins, CpcA and JlbA amino acid starvation-responsive transcription factors, and some elements of sexual development and sporulation was ROS responsive. CONCLUSION: The existence of separate O2(2-), O2*- and GSH/GSSG responsive gene groups in a eukaryotic genome has been demonstrated. Oxidant-triggered, genome wide transcriptional changes should be analyzed considering changes in oxidative stress-responsive physiological conditions and not correlating them directly to the chemistry and concentrations of the oxidative stress-inducing agent. PMID- 16368012 TI - Genome-wide comparative analysis of the IQD gene families in Arabidopsis thaliana and Oryza sativa. AB - BACKGROUND: Calcium signaling plays a prominent role in plants for coordinating a wide range of developmental processes and responses to environmental cues. Stimulus-specific generation of intracellular calcium transients, decoding of calcium signatures, and transformation of the signal into cellular responses are integral modules of the transduction process. Several hundred proteins with functions in calcium signaling circuits have been identified, and the number of downstream targets of calcium sensors is expected to increase. We previously identified a novel, calmodulin-binding nuclear protein, IQD1, which stimulates glucosinolate accumulation and plant defense in Arabidopsis thaliana. Here, we present a comparative genome-wide analysis of a new class of putative calmodulin target proteins in Arabidopsis and rice. RESULTS: We identified and analyzed 33 and 29 IQD1-like genes in Arabidopsis thaliana and Oryza sativa, respectively. The encoded IQD proteins contain a plant-specific domain of 67 conserved amino acid residues, referred to as the IQ67 domain, which is characterized by a unique and repetitive arrangement of three different calmodulin recruitment motifs, known as the IQ, 1-5-10, and 1-8-14 motifs. We demonstrated calmodulin binding for IQD20, the smallest IQD protein in Arabidopsis, which consists of a C terminal IQ67 domain and a short N-terminal extension. A striking feature of IQD proteins is the high isoelectric point (approximately 10.3) and frequency of serine residues (approximately 11%). We compared the Arabidopsis and rice IQD gene families in terms of gene structure, chromosome location, predicted protein properties and motifs, phylogenetic relationships, and evolutionary history. The existence of an IQD-like gene in bryophytes suggests that IQD proteins are an ancient family of calmodulin-binding proteins and arose during the early evolution of land plants. CONCLUSION: Comparative phylogenetic analyses indicate that the major IQD gene lineages originated before the monocot-eudicot divergence. The extant IQD loci in Arabidopsis primarily resulted from segmental duplication and reflect preferential retention of paralogous genes, which is characteristic for proteins with regulatory functions. Interaction of IQD1 and IQD20 with calmodulin and the presence of predicted calmodulin binding sites in all IQD family members suggest that IQD proteins are a new class of calmodulin targets. The basic isoelectric point of IQD proteins and their frequently predicted nuclear localization suggest that IQD proteins link calcium signaling pathways to the regulation of gene expression. Our comparative genomics analysis of IQD genes and encoded proteins in two model plant species provides the first step towards the functional dissection of this emerging family of putative calmodulin targets. PMID- 16368013 TI - Expression of minichromosome maintenance protein 2 as a marker for proliferation and prognosis in diffuse large B-cell lymphoma: a tissue microarray and clinico pathological analysis. AB - BACKGROUND: Minichromosome maintenance (MCM) proteins are essential for the initiation of DNA replication and have been found to be relevant markers for prognosis in a variety of tumours. The aim of this study was to assess the proliferative activity of diffuse large B-cell lymphoma (DLBCL) in tissue microarray (TMA) using one of the minichromosome maintenance proteins (Mcm2) and to explore its potential value to predict prognosis. METHODS: Immunohistochemistry for Mcm2 was performed on TMAs constructed from 302 cases of DLBCL. A monoclonal mouse antibody was used after heat induced antigen retrieval. Mcm2 expression was scored quantitatively. Positivity for Mcm2 was defined as presence of nuclear expression of Mcm2 in greater than or equal to 40 % of tumour cells. A statistical analysis was carried out of the association of Mcm2 and the clinico-pathological characteristics. RESULTS: Mcm2 expression was clearly evident in the nuclei of proliferating non-neoplastic cells and tumour cells. Positivity for Mcm2 was found in 46% (98/211) of analysable cases. A significant correlation existed between Mcm2 expression and presence of bulky disease (p = 0.003). Poor disease specific survival was observed in patients with DLBCL positive for Mcm2 expression in the univariate analysis (p = 0.0424). CONCLUSION: Mcm2 expression can be used to assess tumour proliferation and may be useful as an additional prognostic marker to refine the prediction of outcome in DLBCL. PMID- 16368014 TI - Cyclic Vomiting Syndrome in 41 adults: the illness, the patients, and problems of management. AB - BACKGROUND: Cyclic Vomiting Syndrome (CVS) is a disorder characterized by recurrent, stereotypic episodes of incapacitating nausea, vomiting and other symptoms, separated by intervals of comparative wellness. This report describes the clinical features, co-morbidities and problems encountered in management of 41 adult patients who met the diagnostic criteria for CVS. METHODS: This is a retrospective study of adults with CVS seen between 1994 and 2003. Follow-up data were obtained by mailed questionnaires. RESULTS: Age of onset ranged from 2 to 49 years. The duration of CVS at the time of consultation ranged from less than 1 year to 49 years. CVS episodes were stereotypic in respect of their hours of onset, symptomatology and length. Ninety-three percent of patients had recognizable prodromes. Half of the patients experienced a constellation of symptoms consisting of CVS episodes, migraine diathesis, inter-episodic dyspeptic nausea and a history of panic attacks. Deterioration in the course of CVS is indicated by coalescence of episodes in time. The prognosis of CVS is favorable in the majority of patients. CONCLUSION: CVS is a disabling disorder affecting adults as well as children. Because its occurrence in adults is little known, patients experience delayed or mis-diagnosis and ineffectual, sometimes inappropriately invasive management. PMID- 16368015 TI - Comparison of biomechanical and histological properties in dog carotid arteries injured by neointima or intimal thickening. AB - A general formula (Oka and Azuma's equation) has been rigorously derived for the circumferential wall tension in a hollow cylindrical tube in equilibrium. To evaluate the validity and usefulness of Oka and Azuma's equation, T = P(1) x r(1) - P(2) x r(2) (T, circumferential wall tension; P(1) and P(2), internal and external pressures of the tube; r(1) and r(2), the corresponding internal and external radii), we experimentally investigated changes in circumferential wall tension of noninjured (control) and injured dog common carotid arteries by using a newly developed apparatus with a photo- and X-ray-sensitive image sensor. We also studied histological features of the control and injured arteries with special reference to the relation of biomechanical properties. Two types of animal models with injured arteries--balloon-induced neointima or external collar induced intimal thickening--were adopted in the present study. In the control arteries, the circumferential wall tension was experimentally confirmed to change from negative to positive by an increase in intraluminal pressure ranging from 50 to 180 mmHg. The critical intraluminal pressure that produced 0 dyne/cm of the circumferential wall tension was around 135 mmHg. The activation of arterial smooth muscles caused a significant increase in the critical pressure in the control arteries. In the arteries injured by neointima, the critical intraluminal pressure was significantly lower than that in the control. The activation of smooth muscles also significantly increased the critical pressure in the injured arteries. Histological examination demonstrated the existence of a circumferential neointimal formation along with a shortening of the internal diameter. In other arteries injured by intimal thickening, the circumferential wall tension was always negative at intraluminal pressure ranging from 50 to 180 mmHg. Newly developed structures consisted of elastic and collagen fibers, smooth muscles, and extracellular matrix in the intima and media of the injured arteries. These experimental findings suggest that the circumferential wall tension of dog common carotid arteries has been confirmed experimentally to become negative. We have also concluded that circumferential wall tension calculated with Oka-Azuma's equation may be one of the best parameters for evaluating changes in the biomechanical and histological properties of pathologically injured arteries. PMID- 16368016 TI - Bridging the intention-behaviour 'gap': the role of moral norm. AB - This research examined whether intentions aligned with moral norms better predict behaviour compared with intentions aligned with attitudes. Six data sets predicting behaviours in the health domain (smoking, driving over speed limit, applying universal precautions, exercising) were analysed. Moderated regression analysis indicated that participants whose intentions were more aligned with their moral norm were more likely to perform behaviours compared with participants whose intentions were more aligned with their attitude. However, further analysis indicated that this moderation effect was only present when participants construed the behaviour in moral terms. The findings suggest that the theory of planned behaviour should more clearly acknowledge the importance of internalized norms and self-expectations in the development of one's motivation to adopt a given behaviour. PMID- 16368017 TI - First- and higher-order models of attitudes, normative influence, and perceived behavioural control in the theory of planned behaviour. AB - The present study examined the validity of an augmented version of Ajzen's (1991) theory of planned behaviour (TPB), adopting second-order latent factors to explain relationships between the differentiated components of the theory constructs. This model was tested on data from two independent samples, for exercise and dieting behaviours, respectively. The affective and instrumental attitude, injunctive and descriptive norms, and perceived controllability and self-efficacy constructs all exhibited discriminant validity in the first instance. First- and second-order confirmatory factor analyses exhibited satisfactory fit with the observed data from both samples. There were no substantial differences in the fit indices across the first- and second-order models, and the second-order models exhibited the most optimal parsimony corrected fit indices. The higher-order models could therefore not be rejected on the grounds of inferior fit or parsimony. First- and second-order structural equation models accounted for significant variance in intentions and behaviour. These results corroborate Ajzen's (2002a) and Bagozzi, Lee, and van Loo's (2001) premise that recent augmentations of the TPB that differentiate the model components can be subsumed by global, higher-order factors while still making the distinction at the subordinate level. PMID- 16368018 TI - How the past weighs on the present: social representations of history and their role in identity politics. AB - Socially shared representations of history have been important in creating, maintaining and changing a people's identity. Their management and negotiation are central to interethnic and international relations. We present a narrative framework to represent how collectively significant events become (selectively) incorporated in social representations that enable positioning of ethnic, national and supranational identities. This perspective creates diachronic (temporal) links between the functional (e.g. realistic conflict theory), social identity, and cognitive perspectives on intergroup relations. The charters embedded in these representations condition nations with similar interests to adopt different political stances in dealing with current events, and can influence the perceived stability and legitimacy of social orders. They are also instrumental in determining social identity strategies for reacting to negative social comparisons, and can influence the relationships between national and ethnic identities. PMID- 16368019 TI - Talk about receiving, giving, and taking in radio interviews: 'doing modesty' and 'making a virtue out of necessity'. AB - Acts of giving, receiving, and taking constitute much of our social lives, and talk about these acts is ubiquitous. However, such talk has not been studied in its own right. From radio interviews with persons who have accomplished or experienced something deemed remarkable, I identified two social actions that were accomplished by talk about receiving, giving, and taking: 'doing modesty' and 'making a virtue out of necessity'. Doing modesty was present in interviews with celebrities and was deftly managed through the joint use of contrasts by the participants. Making a virtue out of necessity was present in interviews in which the interviewee's experience of adversity was a prominent focus of talk, and was accomplished through the use of irony. I argue that both actions can serve the dual purposes of putting the interviewee in a good light and holding the attention of the listening audience. This research adds to existing work on social ties by providing another route for understanding how we make use of the exchange of social resources. PMID- 16368020 TI - Gender conflict and worldview defence. AB - Applying social identity and terror management theory assumptions to gender conflict we predicted that mortality salience (MS) would lead to an increase in pro-women attitudes in women and a decrease in these attitudes in men. After a MS versus control manipulation, 32 female and 24 male university students evaluated (fictitious) courses in psychology dealing with and supporting the promotion of women. In accordance with our prediction the results showed a significant interaction between sex and MS, indicating that men and women differed in their judgment only under MS but not in the control condition. Whereas men reacted with an increased negative evaluation of the pro-women courses following MS, women on the other hand showed an increased positive evaluation of the courses. Theoretical and practical implications are discussed. PMID- 16368021 TI - Opposition to redistributive employment policies for women: the role of policy experience and group interest. AB - We examined whether group interest affected ideological beliefs and attitudes towards redistributive policies among men and women. We found that group interest influenced meritocratic and neo-sexist beliefs and support for gender-based affirmative action and comparable worth policies. Men and women differed in their ideological beliefs and support for the redistributive policies only when they had conscious experience with these policies. Those with policy experience expressed policy attitudes that corresponded with their gender group's interests, while those lacking such experience did not. We also noted group interest effects within each gender: men who had conscious experience with the policies expressed more opposition and greater neo-sexism and meritocratic beliefs than did men who were not consciously experienced with these policies. In contrast, consciously experienced women expressed more policy support than did their not consciously experienced counterparts. Overall, our findings indicate that group interest is an important determinant of policy attitudes and related ideological beliefs. PMID- 16368022 TI - The ways that people talk about natural resources: discursive strategies as barriers to environmentally sustainable practices. AB - In this paper, we analyse talk about water and energy use taken from nine interviews with citizens of Perth, Western Australia. Participants' talk offered representations of water as a scarce, shared, natural resource that must not be wasted, whereas talk about energy use focused on the environmental impacts of different technologies for generating electricity, rather than on energy as a consumable resource. Participants accounted for their water-use habits by positioning themselves as caught between a personal desire to conserve water and an (incompatible) social obligation to maintain the appearance of their gardens in keeping with the aesthetic appeal of the suburbs in which they lived. We identify several discursive strategies by which people construct the environmental impact of their actions as minimal or unavoidable. These constitute a barrier to the promotion of more environmentally sustainable practices. Potential implications for environmental policy development are discussed, as are the wider issues associated with the development of 'applied' discourse analysis. PMID- 16368023 TI - Sequential strategies of accommodation: a new method in courtroom. AB - The aim of this research is to show the fruitfulness of a sequential analysis approach applied to the study of the dynamic and sequential character of accommodation strategies proposed by communication accommodation theory (CAT). It was applied to the convergence, divergence, maintenance, and control strategies undertaken in the courtroom during 47 hostile examinations from a single criminal case. Each of the 1,850 question-answer exchanges was classified for the content aspects (types of questions and of answers) and for the modalities of turn taking (interrupting, latching, and pausing) by both lawyer and witness. Log-linear analyses were performed. The main results show that the lawyer and the witness used both accommodation and maintenance strategies, even if the latter are used more by the lawyer. Both participants made use of unimodal and bimodal strategies and tended to reciprocate the behaviour of the interlocutor. The conclusions emphasize the advantage of using the sequential analysis approach to the accommodation strategies of CAT: studying multiple behaviours, identifying the direction and the modalities of the strategies even if they act simultaneously, and providing a means to evaluate the weight of the strategies on the whole and in detail. PMID- 16368024 TI - Clearing the air: the effect of experimenter race on target's test performance and subjective experience. AB - According to stereotype threat theory (Steele, 1997), stereotyped targets under perform on challenging tests, in part because they are worried about being viewed in terms of the negative stereotype that they are intellectually inferior. How then are the negative effects of stereotype threat reduced for stereotyped targets? To examine this issue, a study was conducted to investigate whether stereotype threat's adverse effects are reduced when a Black experimenter administers a verbal test to Black participants. We further examined the question of whether Black participants have a subjective awareness of stereotype threat. Results showed that when a Black experimenter gave a verbal test to Black participants, they did not suffer the typical performance decrements associated with stereotype threat. Additionally, results supported the hypothesis that Black participants have conscious access to the experience of stereotype threat and that this effect is partially mediated by their endorsement of the stereotype. PMID- 16368025 TI - Group-based guilt as a predictor of commitment to apology. AB - Whether the Australian government should officially apologize to Indigenous Australians for past wrongs is hotly debated in Australia. The predictors of support amongst non-Indigenous Australians for such an apology were examined in two studies. The first study (N=164) showed that group-based guilt was a good predictor of support for a government apology, as was the perception that non Indigenous Australians were relatively advantaged. In the second study (N=116) it was found that group-based guilt was an excellent predictor of support for apology and was itself predicted by perceived non-Indigenous responsibility for harsh treatment of Indigenous people, and an absence of doubts about the legitimacy of group-based guilt. National identification was not a predictor of group-based guilt. The results of the two studies suggest that, just as individual emotions predict individual action tendencies, so group-based guilt predicts support for actions or decisions to be taken at the collective level. PMID- 16368026 TI - Self-discrepancies in bipolar disorder: comparison of manic, depressed, remitted and normal participants. AB - OBJECTIVES: To study the role of self-discrepancies in different phases of bipolar disorder (manic-depression). METHOD AND DESIGN: Patients with a diagnosis of bipolar disorder in three groups, currently depressed, currently manic or hypomanic, and currently in remission, together with healthy control participants, were administered a modified version of Higgins' Selves Questionnaire. Consistencies between the self-actual, self-ideal and self-ought representations were calculated, together with consistencies between the self actual representation and the believed views of generalized others about the self. RESULTS: In contrast to all other groups, bipolar depressed patients showed marked discrepancies between their self-actual and self-ideal representations, and between their self-actual and self-ought representations. Manic or hypomanic patients showed higher self-actual:self-ideal consistency than non-patient controls. The differences between the depressed participants and the other groups appeared to be accounted for by their very negative self-actual descriptions. Participants in all four groups showed high levels of consistency between self perceptions and the believed perceptions of others about the self. CONCLUSIONS: These findings confirm that beliefs about the self differ between different phases of bipolar disorder and are consistent with the hypothesis that the manic phase involves active avoidance of discrepancies between the self and self ideals. PMID- 16368027 TI - Attributional lability in depression and paranoia. AB - BACKGROUND: Attributions play an important role in depression and paranoia. However, contrary to most attributional models of psychopathology, there is evidence that attributional style is not a stable trait but is affected by recent experiences. METHOD: Paranoid, depressed, and healthy participants were exposed to a mild stressor in the form of the requirement to complete an anagram task, which included insoluble items. Before and afterwards, they completed measures of attributional style for negative events and a contingency judgment task. RESULTS: Replicating previous findings, paranoid patients initially showed low internality scores for negative events and high judgments of control on the contingency judgment task, whereas the depressed participants showed high internality for negative events and low judgments of control. Following the anagram task, both clinical groups showed increased internality for hypothetical negative events. The paranoid participants also gave increased estimates of control on the contingency judgment task. CONCLUSIONS: Attributional style is more labile in paranoid and depressed patients than in healthy individuals. Implications for attributional theories of psychopathology are discussed. PMID- 16368028 TI - The role of panic in acute dissociative reactions following trauma. AB - OBJECTIVE: 'Peritraumatic dissociation' refers to alterations in awareness and memory for events that occur during and shortly after a traumatic experience. Despite the prevalence of reported peritraumatic dissociation, little is known about the mechanisms that mediate dissociative responses in the initial period after trauma. One theory suggests that peritraumatic dissociation may be a response to elevated arousal and panic symptoms during trauma. This study investigated panic symptoms that occurs at the time of the trauma and their relationship to ongoing dissociation in acute stress disorder. DESIGN: A sample of traumatized people with acute stress disorder or controls were administered a range of psychopathology measures within one month of their trauma. METHOD: Fifty one civilian trauma survivors with either acute stress disorder or no acute stress disorder were administered the acute stress disorder interview, the Impact of Event Scale, the Beck Anxiety Inventory, the Peritraumatic Dissociative Experiences Questionnaire, and the Physical Reactions Scale to index panic reactions during the trauma. RESULTS: Hierarchical regression analysis found that panic reactions during the trauma accounted for nearly half of the variance (47%) of peritraumatic dissociation, and subsequent stress reactions accounted for an additional 3% of the variance. CONCLUSIONS: These findings are consistent with proposals that acute dissociation is associated with panic symptoms that occur during the traumatic experience. Possible mechanisms for the association of panic and dissociation are discussed. PMID- 16368029 TI - Hopelessness and positive and negative future thinking in parasuicide. AB - OBJECTIVES: Hopelessness about the future is a key element in suicidal behaviour. The aim of the present study was to examine possible components of hopelessness, in particular, to contrast positive and negative future thinking and to examine separately number, expectancy, and value of anticipated positive and negative future experiences. DESIGN: A correlational design. METHOD: Repeat parasuicide patients (N = 441) were administered the Beck Hopelessness Scale, the Hospital Anxiety and Depression Scale and the future thinking task, a measure of future positive and negative thinking that assesses number, perceived likelihood, and perceived value of anticipated future positive and negative events. RESULTS: Consistent with predictions, hopelessness correlated more strongly with lack of positive thoughts about the future than it did with presence of negative thoughts. Both positive and negative future thinking showed a relationship to hopelessness over and above their relationships to depression (positive future thinking) and anxiety (negative future thinking). Number and likelihood of positive events and likelihood and value of negative events showed both simple and partial relationships to hopelessness. Number of negative events related to hopelessness but only after the other future thinking variables had been controlled for and value of positive events no longer related to hopelessness after controlling for the other variables. CONCLUSIONS: Hopelessness about the future in suicidal individuals is a multi-faceted construct but lack of positive future thinking is more important than presence of negative future thinking. PMID- 16368030 TI - Self-criticism and major depression: an evolutionary perspective. AB - OBJECTIVES: This study sought to incorporate the personality style of self criticism within an evolutionary framework to help explain its relationship to major depression. It was expected that self-critics would engage in poor social comparisons and have greater feelings of internal entrapment, which are both processes related to depression by evolutionary thinkers. DESIGN: A cross sectional design was employed such that participants were first interviewed and then completed several questionnaires. METHODS: The sample consisted of 146 graduate students who had experienced at least one prior episode of major depression, which was confirmed using the Structured Clinical Interview for DSM IV (SCID). Participants were subsequently administered the Center for epidemiological studies depression scale (CESD), Depressive Experiences Questionnaire (DEQ), Social comparison rating scale (SCRS), and Entrapment scale (ES). RESULTS: Regression analyses revealed that self-criticism significantly predicted internal entrapment and social comparison when controlling for mood and for levels of dependency. Subsequent Structural Equation Modelling (SEM) revealed that a factor of self-reported entrapment and social comparison mediated the effect of self-criticism on the number of previous episodes of depression. CONCLUSION: These findings suggest that part of the reason self-critics are vulnerable to clinical episodes of depression lies in their subjective experience of entrapment and in their negative social comparisons. PMID- 16368031 TI - Bad me paranoia in early psychosis: a relatively rare phenomenon. AB - PURPOSE: The aim of this study was to follow up Trower and Chadwick's (1995) proposal that there are two types of paranoia: poor me paranoia, in which the persecution is believed to be undeserved, and bad me paranoia, in which the persecution is seen as a deserved punishment. METHOD: A cross-sectional design was used. A group of 40 participants with early psychosis, who were experiencing persecutory delusions, were assessed using a semi-structured interview on the content of their beliefs. RESULTS: An extremely low rate of bad me paranoia was found in the sample of participants with early psychosis. CONCLUSIONS: People with persecutory delusions who are in the initial stages of their illness do not tend to believe that they are being justifiably punished. Stigma and depression are put forward as tentative explanations of the development of this evaluative belief with time. PMID- 16368032 TI - A randomized controlled trial of psychological interventions for postnatal depression. AB - OBJECTIVES: First, to establish the efficacy of psychological interventions versus routine primary care for the management of postnatal depression (PND). Secondly, to provide a direct comparison of cognitive-behavioural therapy (CBT) versus counselling and, finally, to compare the relative value of group and individual delivery formats. DESIGN: The study involved 192 depressed women drawn from a large community screening programme in Melbourne, Australia and allocated to cognitive behaviour therapy, counselling, or routine primary care. Baseline and post-intervention measures of depression and anxiety were collected in the form of validated self-report inventories. METHOD: Women were screened in the community and diagnosis of depression confirmed with a standardized psychiatric interview. Interventions were of 12 weeks duration, including three partner sessions, and adhered to a structured manual. RESULTS: Psychological intervention per se was superior to routine care in terms of reductions in both depression and anxiety following intervention. CONCLUSIONS: For those women with PND, psychological intervention is a better option than routine care, leading to clinically significant reduction of symptoms. Counselling may be as effective as group cognitive behaviour therapy. The benefits of psychological intervention may be maximized by being delivered on a one-to-one basis. PMID- 16368033 TI - Reminiscence, regrets and activity in older people in residential care: associations with psychological health. AB - BACKGROUND: Claims have been made of the benefits of reminiscence for older people's psychological health. The study reported here set out to determine whether measures of reminiscence, regrets, and activity participation would be associated with psychological health in older people, when age, dependency, self reported health, and social well-being were controlled. DESIGN: Cross-sectional interview-based questionnaire survey. METHODS: Older people living in residential and nursing homes (N = 142) completed questionnaires that determined current levels of reminiscence, activity participation, and psychological health, while care staff recorded the participants' observed affect over the previous 2-week period. RESULTS: Multivariate analyses showed significant associations between reminiscence frequency, reminiscence enjoyment, and regrets, and psychological health outcomes, while controlling for age, dependency, self-reported health, and social well-being. While reminiscence enjoyment was associated with positive psychological health, high frequency of reminiscence and the presence of regrets were associated with negative psychological health. CONCLUSIONS: The study findings are discussed with reference to Eriksonian theory, reminiscence functions, and the potential for reminiscence in psychological therapy for older people. PMID- 16368034 TI - The childhood experience of care and abuse questionnaire (CECA.Q): validation in a community series. AB - BACKGROUND: Childhood neglect and abuse, as measured by retrospective interview, is highly predictive of psychiatric disorder in adult life and has an important role in aetiological models. However, such measures are labour-intensive, costly, and thus restricted to relatively modest sample sizes. A compact self-report assessment of childhood experience is invaluable for research screening purposes and large-scale survey investigation. METHOD: A self-report questionnaire (CECA.Q) was developed to mirror an existing validated interview measure: the childhood experience of care and abuse (CECA). The questionnaire assessed lack of parental care (neglect and antipathy), parental physical abuse, and sexual abuse from any adult before age 17. A high-risk series of 179 London women were interviewed using the CECA together with the PSE psychiatric assessment, and completed the CECA.Q at later follow-up. Repeat CECA.Qs were returned for 111 women and 99 women additionally completed the parental bonding instrument (PBI; Parker, Tupling, & Brown, 1979). RESULTS: Satisfactory internal scale consistency was achieved on the CECA.Q for antipathy (alpha = .81) and neglect (alpha = .80) scales. There was satisfactory test-retest for both care and abuse scales. Significant associations were found between CECA.Q scales and the parallel interview scales with cut-offs determined for high sensitivity and specificity. CECA.Q neglect and antipathy scales were also significantly related to PBI parental care. CECA.Q scales were significantly related to lifetime history of depression. Optimal cut-off scores revealed significant odds ratios (average of 2) for individual scales and depression. When indices were compiled to reflect peak severity of each type of adversity across perpetrator, odds-ratios increased (average 3). A dose-response effect was evident with the number of types of neglect/abuse and rate of lifetime depression. CONCLUSION: The CECA.Q shows satisfactory reliability and validity as a self-report measure for adverse childhood experience. The merits of having parallel questionnaire and interview instruments for both research and clinical work are discussed. PMID- 16368035 TI - Reduced autobiographical memory specificity as an avoidant coping style. AB - OBJECTIVE: Previous studies suggest that reduced autobiographical memory specificity might serve as a way of regulating affect. This study examined the relationship between autobiographical memory specificity and avoidant coping. METHOD: Sixty students completed the Autobiographical Memory Test (AMT). Different types of avoidant coping were assessed using questionnaires: the Acceptance and Action Questionnaire, the Cognitive-Behavioural Avoidance Scale and the White Bear Suppression Inventory. RESULTS: For all questionnaires, a significant correlation was observed between the number of specific responses on the AMT and an avoidant coping style. More avoidance was associated with fewer specific answers. These correlations remained when corrected for number of omissions. CONCLUSION: The results provide further evidence for the affect regulating quality of autobiographical memory specificity. PMID- 16368036 TI - Self-defining memories in post-traumatic stress disorder. AB - OBJECTIVE: This study investigated the relationship between trauma survivors' goals and retrieval of self-defining memories in post-traumatic stress disorder (PTSD). METHODS: Civilian trauma survivors with PTSD, trauma survivors with no PTSD and non-trauma-exposed control participants (N = 49) provided autobiographical memories of events that they believe shaped who they are. Participants also provided details about their major personal goals. RESULTS: Participants with PTSD reported more self-defining memories that were trauma related, negative valence and from adult years than non-PTSD and control participants. Further, retrieval of trauma-related self-defining memories was strongly associated with reporting personal goals that were related to traumatic experiences. CONCLUSIONS: These findings are discussed in terms of the proposition that trauma survivors' current concerns may direct retrieval of trauma-related memories. PMID- 16368037 TI - Prevalence of vitamin D inadequacy in osteoporotic hip fracture patients in London. AB - BACKGROUND: It is well established that vitamin D levels are suboptimal in the elderly and that adults with fragility fracture are more likely to have serum vitamin D levels either lower than those of control patients of similar age, or below the normal range. OBJECTIVES: To investigate the prevalence of vitamin D inadequacy in an elderly population with hip fractures from London (UK) and compare levels with data previously presented from Glasgow (UK). RESEARCH DESIGN AND METHODS: A retrospective patient audit was carried out over a 17-month period (September 2003-January 2005). Patient records were searched for hip fracture admissions and cross matched with vitamin D analysis carried out within 3 days of the hip fracture admission. The resulting records were hand searched to exclude patients with a hip fracture resulting from high impact/trauma. RESULTS: There were data for 103 hip fracture patients, 79.6% of the patients were women (n = 82). The mean age at the time of fracture was 73.4 years, 100% were aged 60 years or over and 41% were aged 75 years or over. Around 20% of the patients were receiving supplementation with calcium and/or vitamin D and were not excluded from the analysis. The mean vitamin D level was 32.1 nmol/L (12.9 ng/mL), SD = 19.4 (7.8), however, it is likely that the true mean is lower since in approximately 15% of cases vitamin D levels were reported as < 12.5 nmol/L, but were transcribed at 12.5 nmol/L in order to allow a numerical value to be calculated. Ninety-nine per cent of patients had a vitamin D level < 80 nmol/L, 94.2% < 70 nmol/L and 81.6% < 50 nmol/L. There were no significant differences by patient age or sex, however, there were significant seasonal differences in vitamin D. In the year from September 2003 to August 2004, 82.8% of summer admissions had vitamin D levels < 70 nmol/L compared with 98.0% in winter (p = 0.04). Mean vitamin D levels in the 30 patients with parathyroid hormone (PTH) levels above the reference range were significantly lower than levels in the 71 patients within the range: mean 19.9 nmol/L, SD = 16.2 versus mean 37.5 nmol/L, SD = 18.5 (p < 0.0001). Furthermore, 50% of the patients with PTH levels above the reference range had vitamin D levels < 12.5 nmol/L, reflecting extremely low levels of vitamin D. CONCLUSIONS: This study confirms almost universal vitamin D inadequacy among 103 patients admitted to hospital with hip fracture in London, although the prevalence of inadequacy is slightly lower than that seen in a similar study carried out in Glasgow. PMID- 16368038 TI - Patient preference for once-monthly ibandronate versus once-weekly alendronate in a randomized, open-label, cross-over trial: the Boniva Alendronate Trial in Osteoporosis (BALTO). AB - OBJECTIVE: Ibandronate, a potent nitrogen-containing bisphosphonate, can be administered with extended interval dosing. Patient preferences were assessed for once-monthly versus once-weekly bisphosphonate treatment using a previously developed, open-label, cross-over trial design. RESEARCH DESIGN AND METHODS: This was a 6-month, prospective, randomized, open-label, multi-center study with a two period and two-sequence cross-over treatment design. After screening, eligible patients (postmenopausal women with osteoporosis) were randomized to once-monthly ibandronate 150 mg followed by once-weekly alendronate 70 mg for a total of 6 months (Sequence A) or once-weekly alendronate followed by once-monthly ibandronate for a total of 6 months (Sequence B). The primary objective was to evaluate patient-reported preference for either the once-monthly ibandronate regimen or the once-weekly alendronate regimen based on responses to a preference questionnaire. RESULTS: A total of 342 patients were enrolled into this study (Sequence A, 170; Sequence B, 172). In the primary analysis of patient preference, 71.4% of women selected once-monthly ibandronate and 28.6% of women selected once-weekly alendronate. Overall, 66.1% preferred the once-monthly ibandronate regimen to the once-weekly alendronate regimen (26.5%) and 7.4% of participants stated no preference for either regimen. The preference rate for once-monthly ibandronate was statistically significant (p < 0.0001). 'Ease of following a treatment regimen for a long time' was the most common reason given for patient preference for both the once-monthly ibandronate (61%, 169/276) and once-weekly alendronate (25%, 70/276) regimens. Additionally, 17% (47/276) of patients who preferred once-monthly ibandronate chose 'it is easier to tolerate side effects' as did 4.3% (12/276) of patients who preferred alendronate. Significantly more women found once-monthly ibandronate to be more convenient (p < 0.0001). CONCLUSIONS: Significantly more women with postmenopausal osteoporosis preferred once-monthly ibandronate therapy to once-weekly alendronate therapy, and found the once-monthly regimen to be more convenient. Ease of following a treatment regimen for a long time was the most common reason given for the patients' preferences. PMID- 16368039 TI - Anticoagulation in patients with non-valvular atrial fibrillation: an evaluation of stability and early factors that predict longer-term stability on warfarin in a large UK population. AB - OBJECTIVE: To determine the proportion of patients with non-valvular atrial fibrillation (NVAF) treated with warfarin that achieved a 6-month period within the target INR range (stability). To then evaluate any associations between stability and outcome and to determine whether stability can be predicted by clinical factors at an early stage in warfarin treatment. METHODS: This study was a record linkage study in 1513 patients with NVAF treated with warfarin for a minimum of 6-months, carried out in a large UK population. The main outcome measures were stability (defined as six months within the target INR range [2.0 3.0]), thromboembolic and bleeding event rates and mortality. Secondary outcome measures were the predictive value of baseline characteristics and other treatment variables. RESULTS: Stability was achieved in 52% of the study group. Standardised mean survival was significantly higher in the group who achieved stability (Delta = 16.91 months, p < 0.001) with a hazard ratio of 4.36 (p < 0.001). The stable group had a lower rate of both thromboembolic events (0.8% vs. 2.3% per patient year) and bleeds recorded on inpatient diagnoses (0.4% vs. 1.2% per patient year). Failure to achieve stable control was associated with age (Odds Ratio [OR] 1.011 (95% Confidence Interval [CI] 1.001-1.021)) and morbidity at baseline (OR 1.015; 95% CI 1.007-1.022). An increase in mean time between visits (OR 0.939; 95% CI 0.926-0.952) and the percentage time in range (OR 0.889; 95% CI 0.879-0.900) was associated with a decrease in the chance of instability. Greater variability in INR was also associated with a failure to achieve stability (OR 1.518; 95% CI 1.427-1.615). Receiver Operator Characteristic (ROC) analysis using data from the first three months of treatment demonstrated good discrimination of stability using age and morbidity at baseline and percentage time in range and frequency of visits during the first three months of treatment (area under curve [AUC] 0.780; standard error [SE] 0.012; 95% CI 0.757-0.803). CONCLUSIONS: Many patients never achieved a period of 6-months stability and were at increased risk of thromboembolic events and bleeds. Age, morbidity at baseline and variability of INR control in the first three months could be used to predict instability using warfarin. This study infers that patients should be treated more intensively in the early stages of warfarinisation in order to improve outcome. PMID- 16368040 TI - Short-term economic impact associated with occupational needlestick injuries among acute care nurses. AB - PURPOSE: Recent survey data have reported the incidence rate of needlestick injuries (NIs) and NIs which draw blood sustained by nurses caring for patients with diabetes in an in-patient hospital setting. The purpose of this study was to deduce the potential short-term annual economic impact resulting from such NI, and to project the potential national economic burden of NI among this population of health care workers (HCWs). METHODS: Data were obtained from a recently published, IRB-approved, Internet-based survey in which nurses routinely treating patients with diabetes self-reported outcomes of their experience with NI (N = 400). A micro-costing approach was adopted. Direct costs comprised post-exposure testing (PET) for infection, post-exposure health care services utilization, and NI-induced post-exposure prophylactic (PEP) drug utilization. Indirect costs were derived from missed workdays and associated lost productivity. These data were combined with data related to the national epidemiology and total incidence of NIs among HCWs and risk-associated populations to project the national burden. RESULTS: Among 400 nurses, 110 sustained at least one NI in the past year, with 73 punctures drawing blood. The ensuing total short-term costs of these NIs were calculated to range from 25,896 US dollars to 36,066 US dollars. Indirect costs accounted for 44-62% of this total cost. Average short-term costs per NI ranged from 145 to 201 US dollars, and average short-term costs of NI per injured nurse ranged from 235 to 328 US dollars. Assuming mean values from published literature on the incidence and distribution of NI among nursing populations, an annual national burden of 65 million US dollars was calculated for costs in the immediate period following NI. CONCLUSIONS: These data suggest substantial economic burden immediately following NI on a national and individual hospital level occurring among acute-care nurses treating patients with diabetes. Long term treatment costs would add to the overall economic burden. PMID- 16368041 TI - Use of daptomycin to treat drug-resistant Gram-positive bone and joint infections. AB - OBJECTIVE: Drug-resistant, Gram-positive bacteria are a growing concern in treating bone and joint infections, including osteomyelitis. This report describes the experience in a series of cases of the use of a novel antibiotic, daptomycin, for the treatment of bone and joint infections. RESEARCH DESIGN AND METHODS: This retrospective analysis included patients from two medical centers diagnosed with Gram-positive bone and joint infections and treated with daptomycin. RESULTS: A total of 10 patients were included in this report, of which nine received previous antibiotic therapy, including vancomycin, linezolid, and quinupristin/dalfopristin. Methicillin-resistant Staphylococcus aureus was isolated from eight patients while the remaining patients were infected with enterococci or streptococci. All patients initially resolved the infection while undergoing daptomycin treatment and were discharged from the hospital. One patient was switched to ampicillin (after receiving daptomycin for 4 days) once the infection was identified due to vancomycin-susceptible enterococcus. However, one patient was readmitted after 18 days due to a clinical relapse, possibly caused by under-dosing of daptomycin. CONCLUSION: Eight out of nine patients who received daptomycin for at least 8 days were successfully treated with the agent for Gram-positive bone and joint infections. Daptomycin was found to be well tolerated, even up to 44 days of treatment. PMID- 16368042 TI - High prevalence of low HDL-cholesterol in a pan-European survey of 8545 dyslipidaemic patients. AB - OBJECTIVE: Low HDL-cholesterol is a strong independent cardiovascular risk factor recognized as a therapeutic target in recent guidelines. The Pan-European Survey on HDL-cholesterol collected data on plasma lipid profiles from 8545 dyslipidaemic patients in the care of 1339 specialist physicians in 11 European countries. The main objective was to obtain a reliable estimation of the prevalence of low HDL (< 1.03 mmol/L [< 40 mg/dL] in men; < 1.29 mmol/L [< 50 mg/dL] in women). RESEARCH DESIGN AND METHODS: Eligible patients were aged >or= 18 years and had received diet and exercise plus pharmacologic lipid-modifying treatment for >or= 3 months, or had serum cholesterol >or= 5.18 mmol/L (>or= 200 mg/dL) and/or serum triglycerides >or= 2.03 mmol/L (>or= 180 mg/dL) despite >or= 3 months of diet and exercise. RESULTS: The survey population was overweight (mean body mass index 29.0 kg/m2), with a high prevalence of sedentary lifestyle (68%), type 2 diabetes (45%), hypertension (72%) and coronary heart disease (45%). Lipid-modifying treatment, received by 85% of patients, included lifestyle modification (85%) and/or lipid-lowering drugs (85% received a statin). The prevalence of low HDL-cholesterol despite lipid-modifying treatment was 40% (women) and 33% (men). Very low HDL-cholesterol (< 0.90 mmol/L [< 35 mg/dL]) occurred in 14% of treated patients, with similar prevalence in the subgroup of patients not receiving such treatment. Hypertriglyceridaemia (> 1.69 mmol/L [> 150 mg/dL]) occurred in 57% of men not under lipid-modifying treatment and in 47% of men receiving such treatment; corresponding values in women were 48% and 43%. Both low HDL-cholesterol and hypertriglyceridaemia occurred in 26% of men and 27% of women who were not receiving lipid-modifying treatment and in 21% of men and 25% of women receiving lipid-modifying treatment. CONCLUSIONS: Low HDL cholesterol and hypertriglyceridaemia are, therefore, common among European patients treated for dyslipidaemia. Clearly, physicians need to focus more of their attention on this major risk factor and to consider adequate treatment when indicated. PMID- 16368043 TI - Short and longer term outcomes of a diabetes disease management program in underserved Latino patients. AB - OBJECTIVE: We developed a Diabetes Management Program (DMP) located within a county health center for poor, primarily Latino patients. The purpose of this study is to document if our program can achieve global risk reduction in our population, and whether or not this improvement can be sustained through on-going follow-up in primary care. RESEARCH DESIGN AND METHODS: This is an observational study, using a database into which patient information is entered weekly. Information from the database is used to compare patients at baseline, at discharge from the program and then after a year or more of follow-up in primary care. MAIN OUTCOMES MEASURES: Change in A1C, low density lipoprotein (LDL) cholesterol, blood pressure (BP) levels; use of cardiovascular disease (CVD) modifying medications. RESULTS: 171 patients had data available at baseline, discharge and annual follow-up. Baseline A1C is 8.5 +/- 1.9% (mean +/- SD), with a fall to 7.4 +/- 1.2% (p < 0.001) at discharge. After 1 year in primary care the A1C increased back to baseline (8.3 +/- 1.7%), without change in diabetes medications. LDL cholesterol and BP levels are reduced by discharge, and these improvements persisted at the annual follow-up. Additionally, patients continued with CVD risk modifying therapy (statins, angiotensin converting enzyme inhibitors/angiotensin receptor blockers, aspirin) at their annual visits. Limitations to this study include the small number of patients who returned for their annual examination, the lack of a matched control group for comparison and the use of an existing database. CONCLUSIONS: Persistent change in CVD risk factors can be made through diabetes disease management; changes in glucose control, however, are more difficult to sustain in spite of continued use of medication. Further research must be undertaken to help improve glycemic outcomes over time. PMID- 16368044 TI - Influence of switching to travoprost on intraocular pressure of uncontrolled chronic open-angle glaucoma patients compliant to previously-used topical medication. AB - OBJECTIVE: To investigate the influence of switching to travoprost on intraocular pressure (IOP) of chronic open-angle glaucoma (COAG) patients. RESEARCH DESIGN AND METHODS: Multicentre, open-label, non-comparative, 12-week, phase IV study conducted at 10 academic and hospital centres in Hungary. Patients' compliance to use of the pre-study medication was confirmed at a visit 10 days before the baseline measurements, and compliance was monitored throughout the study period. RESULTS: Of the 203 COAG patients three (1.48%) ceased travoprost medication due to ocular hyperaemia, and one subject was lost from follow-up. Self-reported compliance was optimal except for two patients. For the per-protocol analysis 197 patients were evaluable. IOP of the 37 per-protocol patients receiving additional travoprost medication decreased from 23.1 +/- 3.2 mmHg (mean +/- SD) to 17.3 +/- 2.6 mmHg at week 12 (p < 0.001). Switching the 121 per-protocol patients from latanoprost to travoprost IOP decrease from 20.8 +/- 3.5 mmHg to 17.7 +/- 2.4 mmHg (p < 0.001). IOP of the 11 patients switched from topical nonselective beta blockers to travoprost decreased from 20.1 +/- 2.1 mmHg to 15.7 +/- 1.5 mmHg (p < 0.001). For the whole per-protocol population (n = 197) IOP decreased from 21.0 +/- 3.4 mmHg to 17.4 +/- 2.4 mmHg (p < 0.001). Defining responders as having an IOP decrease > 2.0 mmHg or >or= 5 mmHg at week 12, the responder rate was respectively 62.9% or 31% for the total study population; 86.5% or 54.1% when travoprost was added to the established therapy; 54.5% or 24.0% if latanoprost was switched to travoprost; and 90.9% or 36.4% for those who switched from beta blockers. CONCLUSION: Travoprost provided a clinically and statistically significant IOP decrease in uncontrolled COAG patients whose self-reported compliance to their previous topical medication was optimal. Our results suggest that the IOP reduction found after switching to travoprost is not explainable by improved compliance due to the clinical study situation. PMID- 16368045 TI - Efficacy of a new aluminium salt thermophobic foam in the treatment of axillary and palmar primary hyperhidrosis: a pilot exploratory trial. AB - BACKGROUND: Primary or idiopathic hyperhidrosis (PH) is a disorder of excessive eccrine sweating glands that mainly affects the axillae and the palms. The treatment options for PH involve a range of topical or systemic medication and/or surgical invasive techniques. The common topical treatments are aluminium salts which act by blocking the duct of the eccrine gland or by atrophying the secretory cells. Recently, a new low-residue thermophobic foam formulation (VersaFoam, Mipharm Spa, Milan, Italy), containing 20% of an aluminium salt (sesquichlorhydrate), has been developed. The foam is easy to apply especially in hairy body sites. OBJECTIVE: to evaluate the efficacy and the tolerability of the new aluminium salt foam in the treatment of axillary and palmar PH. PATIENTS AND METHODS: Twenty patients were enrolled in a single-centre, open-label follow-up study. The Minor test score (range 0-3) and the Dermatology Life Quality Index (DLQI), were used to evaluate the amount of sweating and the impact on quality of life. The foam was applied to dry, clean skin, every night during the first week of treatment, and three times a week during the second week of treatment. Evaluation of the results was performed at baseline and at 7 and 15 days after treatment. Patients were monitored throughout the study for adverse events. RESULTS: All of the 20 enrolled patients completed the study. The foam resulted in a significant reduction of the Minor score in comparison with baseline values in both the axillary (p = 0.0002) and palm regions (p = 0.0047). By the end of treatment (day 15) the foam had reduced the amount of sweating in the axillae and palm regions by 50% (Minor score: 4.1 vs. 8.1) and 53% (Minor score: 4.0 vs. 8.5), respectively. Use of the foam showed a positive impact in the DLQI for patients with axillary but not palm hyperhidrosis. No side effects were reported during the study duration by the patients. CONCLUSION: The new foam has been shown to be an effective topical treatment in reducing sweating in patients with axillary and palm PH. This formulation is well tolerated in the short term. Further studies are warranted to evaluate the efficacy and safety in the medium and long term. PMID- 16368046 TI - The effect of raloxifene therapy on the risk of new clinical vertebral fractures at three and six months: a secondary analysis of the MORE trial. AB - OBJECTIVE: Raloxifene treatment (60 mg/day) significantly decreases the risk of new clinical vertebral fractures by 68% at 1 year compared with placebo. The objective of the present analysis is to evaluate the effects of raloxifene on the incidence of new clinical vertebral fractures at 3 and 6 months after initiation of treatment. RESEARCH DESIGN AND METHODS: A double-blind, randomized, placebo controlled, 4-year Multiple Outcomes of Raloxifene Evaluation (MORE) trial was conducted in 180 study centers. Postmenopausal women with osteoporosis (N = 7705) were randomized to placebo, or raloxifene at 60 or 120 mg/day. MAIN OUTCOME MEASURES: Vertebral radiographs were obtained when patients reported symptoms suggestive of vertebral fracture at or between clinic visits, which were held at 3 and 6 months, and every 6 months thereafter. If a new adjudicated fracture was found, this was considered as a clinical vertebral fracture. The analyses included all randomized patients with a baseline and at least one follow-up radiograph (n = 6828). RESULTS: One woman treated with raloxifene 60 mg/day (n = 2259) and 10 in the placebo group (n = 2292) had a clinical vertebral fracture in the first 6 months, resulting in a 90% relative risk (RR) reduction [RR 0.10 (95% CI 0.01, 0.63)] and a 0.39% absolute risk reduction (ARR). Similar results were observed with raloxifene 120 mg/day at 6 months. When the raloxifene groups were pooled, a significant (p = 0.034) decrease in clinical vertebral fracture risk [RR 0.20 (95% CI 0.03, 0.90), ARR 0.17%] was seen as early as 3 months. CONCLUSION: The risk of new clinical vertebral fractures was reduced after 3 or 6 months of raloxifene. PMID- 16368047 TI - Evaluation of the bioequivalence of two transdermal fentanyl systems following single and repeat applications. AB - OBJECTIVE: Transdermal delivery of fentanyl has potential benefits over slow release morphine, being largely preferred by patients owing to the combination of effective pain relief, a good safety profile and easy, pain-free dosing. The new drug-in-adhesive Durogesic D-TRANS fentanyl Matrix Delivery System (DDTDF) has improved pharmaceutical characteristics and patient acceptability compared to the original Durogesic transdermal reservoir system (fentanyl transdermal reservoir), whilst still providing reliable and consistent delivery of fentanyl. The bioequivalence of these two systems was evaluated in two studies. RESEARCH DESIGNS AND METHODS: Eighty healthy volunteers received single (72 h) or multiple (288 h) applications of DDTDF and the transdermal reservoir system (100 microg/h) in two separate randomised, crossover bioequivalence studies. Bioequivalence was assessed by calculating the ratio of least squares means based on log-transformed data following single system application and at steady-state during the fourth application. RESULTS: Both transdermal systems were bioequivalent with respect to all tested pharmacokinetic parameters. Inter-subject variability was comparable between the two systems and was greater than intra-subject variability. Transdermal delivery was well tolerated in both groups. CONCLUSIONS: The pharmacokinetic results demonstrate that DDTDF is bioequivalent to the original fentanyl transdermal reservoir system after single and multiple applications. PMID- 16368048 TI - Comparative characterisation of a commercial human chorionic gonadotrophin extracted from human urine with a commercial recombinant human chorionic gonadotrophin. AB - BACKGROUND: A commercial preparation of recombinant human chorionic gonadotrophin (r-hCG, Ovitrelle) was launched in 2001. Generally, hCG is available in two formats: human chorionic gonadotrophin (u-hCG), derived from the urine of pregnant females, and r-hCG produced by DNA based biotechnology. METHOD: The analytical characteristics of a highly purified u-hCG (Gonasi HP) were assessed and compared, for the first time, with the recombinant derived r-hCG (Ovitrelle). Gonasi HP is produced by extracting and purifying hCG from urine to obtain a specific bioactivity of 5000 IU/mg protein. Ovitrelle is produced via a recombinant derived mammalian cell line and purified to obtain a specific activity of 26 000 IU/mg. RESULTS AND CONCLUSION: It has been documented that commercially available u-hCG preparations can contain a number of urine derived protein contaminants as well as hCG related metabolites. This is also the case for Gonasi HP, where hCG related molecules and other proteins were found to be present, including epidermal growth factor (EGF) and eosinophil derived neurotoxin (EDN). It was also demonstrated that this preparation contained high levels of oxidised hCG. r-hCG was confirmed to be essentially intact hCG, free from contaminant proteins and with very low levels of oxidised hCG. PMID- 16368049 TI - Predictors of high-cost managed care patients with acute coronary syndrome. AB - OBJECTIVE: To develop predictive models of high-cost acute coronary syndrome (ACS) patients using demographic, disease, and treatment characteristics. STUDY DESIGN: This was a retrospective, administrative claims analysis utilizing pharmacy, medical, and eligibility data from a large US managed care organization. METHODS: ACS was defined by ICD-9 codes for unstable angina (UA) or acute myocardial infarction (AMI). New onset patients (without ACS claims) in the prior six months were identified for the time period 07/01/99-06/30/01, and followed up to 12 months, health plan disenrollment, or death. Cost was measured as that incurred during the initial episode plus subsequent follow-up or during the subsequent follow-up only. Patients were dichotomized as high-cost (top 20%) or low-cost (bottom 80%), based on total costs. Logistic regression was used to examine the association for being classified as high-cost. RESULTS: A total of 13 731 patients were included: 51.7% with UA, 39.6% with AMI and 8.7% with both UA and AMI. The mean age was 54.2 years and 68.2% were male. A number of co morbidities (hypertension, diabetes, heart failure, etc.) predicted high-cost patients. Among medications, prior ACE inhibitor use predicted high-cost patients. While revascularization procedures, in general, were strong predictors of high-cost, revascularization during the index ACS episode (opposed to revascularization during the follow-up) decreased the odds of being high-cost (odds ratio [95% CI] 0.615 [0.506-0.748]). CONCLUSION: High-cost patients with new onset ACS can be predicted by some characteristics, but many of these characteristics are non-modifiable co-morbidities. Payers and providers may find opportunities for clinical and cost-saving interventions for these patients. PMID- 16368050 TI - Extended adjuvant endocrine therapy of early breast cancer. AB - Women who are diagnosed with early breast cancer remain at considerable risk of recurrence over the next several decades, even if their tumors were small and lymph nodes were negative, and despite receiving standard adjuvant therapy. A majority of breast cancers are hormone (estrogen) receptor-positive and amenable to endocrine therapy, and for those women five years of the selective estrogen receptor modulator tamoxifen is standard therapy. Longer treatment of node negative patients with tamoxifen may reduce survival benefits, however, possibly due to tamoxifen resistance and emerging receptor agonist activity of that drug. Aromatase inhibitors, which indirectly prevent estrogen stimulation of breast cancer by suppressing whole-body estrogen synthesis in post-menopausal women, are being investigated as alternative, or complementary, therapy to adjuvant tamoxifen in those women: as an alternative to five years of tamoxifen, sequenced with two to three years of tamoxifen, or following five years of tamoxifen. The strategy to extend the benefits of adjuvant therapy beyond a standard course of tamoxifen, using the aromatase inhibitor letrozole, was explored in a large trial, MA.17. Compared with women who received placebo, those who were treated with letrozole experienced a significant 43% reduction in their residual risk of recurrence. This effect was seen regardless of nodal status. Based on the long term risk of most women with early breast cancer and the MA.17 trial results, the extended adjuvant letrozole may benefit many of those women who are disease-free after five years of tamoxifen. This review is based on a literature search of databases including MEDLINE/PubMed, San Antonio Breast Cancer Symposium, and the Annual Meeting of the American Society of Clinical Oncology, up to and including August 2005, with information selected for its relevance to adjuvant therapy of breast cancer with endocrine therapy only. PMID- 16368051 TI - Effect of orlistat, micronised fenofibrate and their combination on metabolic parameters in overweight and obese patients with the metabolic syndrome: the FenOrli study. AB - BACKGROUND: Obesity is becoming increasingly common worldwide and is strongly associated with the metabolic syndrome (MetS). MetS is considered to be a cluster of risk factors that increase the risk of vascular events. OBJECTIVE: In an open label randomised study (the FenOrli study) we assessed the effect of orlistat and fenofibrate treatment, alone or in combination on reversing the diagnosis of the MetS (primary end-point) as well as on anthropometric and metabolic parameters (secondary end-points) in overweight and obese patients with MetS but no diabetes. METHODS: Overweight and obese patients (N = 89, body mass index (BMI) > 28 kg/m2) with MetS [as defined by the National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III criteria] participated in the study. Patients were prescribed a low-calorie low-fat diet and were randomly allocated to receive orlistat 120 mg three times a day (tid) (O group), micronised fenofibrate 200 mg/day (F group), or orlistat 120 mg tid plus micronised fenofibrate 200 mg/day (OF group). Body weight, BMI, waist circumference, blood pressure, serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL C), high-density lipoprotein cholesterol (HDL-C), non-HDL-C, triglyceride, creatinine (SCr) and uric acid (SUA) levels, as well as homeostasis model assessment (HOMA) index and liver enzyme activities were measured at baseline and after 3 months of treatment. RESULTS: Of the 89 patients enrolled, three (one in each group) dropped out during the study due to side effects. After the 3-month treatment period, 43.5% of patients in the O group, 47.6% in the F group and 50% in the OF group no longer met the MetS diagnostic criteria (primary end-point, p < 0.0001 vs. baseline in all treatment groups). No significant difference in the primary end-point was observed between the three treatment groups. Significant reductions in body weight, BMI, waist circumference, blood pressure, TC, LDL-C, non-HDL-C, triglyceride and SUA levels, as well as gamma-glutamyl transpeptidase activity and HOMA index were observed in all treatment groups. In the OF group a greater decrease in TC (-26%) and LDL-C (-30%) was observed compared with that in the O and F groups (p < 0.01) and a more pronounced reduction of triglycerides ( 37%) compared with that in the O group (p < 0.05). SUA levels and alkaline phosphatase activity decreased more in the F and OF groups compared with the O group (p < 0.05). Moreover, SCr significantly increased and estimated creatinine clearance decreased in the F and OF groups but they were not significantly altered in the O group (p < 0.01 for the comparison between O and either F or OF groups). Glucose (in groups O and OF), as well as insulin levels and HOMA index (in all groups), were significantly reduced after treatment (p < 0.05 vs. baseline). CONCLUSION: The combination of orlistat and micronised fenofibrate appears to be safe and may further improve metabolic parameters in overweight and obese patients with MetS compared with each monotherapy. PMID- 16368052 TI - Cost-minimization analysis of treprostinil vs. epoprostenol as an alternate to oral therapy non-responders for the treatment of pulmonary arterial hypertension. AB - INTRODUCTION: Idiopathic pulmonary arterial hypertension (IPAH) is associated with substantial morbidity and mortality. Treprostinil was compared to epoprostenol for the economic impact of treating IPAH patients who failed or were not candidates for bosentan. METHODS: The model was a cost-minimization analysis, assuming clinical equivalence was achieved by proper dosing of both drugs, in terms of survival and surrogate measures. Two theoretical cohorts of 270 patients were treated with subcutaneous treprostinil and intravenous epoprostenol, and were evaluated over 3 years using a spreadsheet model. Annual survival rates were estimated for the cohorts so that at endpoint 114 (42%) patients survived in both groups. The model utilized resource valuation data for medication and supply costs from Medicare; hospital, consultation, surgical, and diagnostic procedural fees from North Carolina hospitals; and costs to treat adverse events from published sources. Costs were obtained from standard lists and were presented as 2003 US dollars, discounted at 3%. Sensitivity analyses were performed testing all model uncertainties. RESULTS: In the base case analysis, treprostinil demonstrated savings of 22,701 US dollars and 37,433 US dollars per patient over 1- and 3-year time horizons, respectively. The greatest savings came from reduced or minimal hospitalizations attributed to the dose titration and treatment of adverse events, such as sepsis, associated with epoprostenol and its delivery system. Probabilistic sensitivity analyses resulted in average 3-year cost savings of 41,051 US dollars (Standard Deviation = 13,902 US dollars) per patient. CONCLUSIONS: By initiating and continuing treatment with treprostinil over a 3-year period, the economic burden associated with IPAH may be reduced compared to treatment with epoprostenol. The greatest saving with treprostinil was attributed to decreased sepsis. PMID- 16368053 TI - The economic burden of schizophrenia in Canada in 2004. AB - OBJECTIVE: To estimate the financial burden of schizophrenia in Canada in 2004. METHODS: A prevalence-based cost-of-illness (COI) approach was used. The primary sources of information for the study included a review of the published literature, a review of published reports and documents, secondary analysis of administrative datasets, and information collected directly from various federal and provincial government programs and services. The literature review included publications up to April 2005 reported in MedLine, EMBASE and PsychINFO. Where specific information from a province was not available, the method of mean substitution from other provinces was used. Costs incurred by various levels/departments of government were separated into healthcare and non healthcare costs. Also included in the analysis was the value of lost productivity for premature mortality and morbidity associated with schizophrenia. Sensitivity analysis was used to test major cost assumptions used in the analysis. Where possible, all resource utilization estimates for the financial burden of schizophrenia were obtained for 2004 and are expressed in 2004 Canadian dollars (CAN dollars). RESULTS: The estimated number of persons with schizophrenia in Canada in 2004 was 234 305 (95% CI, 136 201-333 402). The direct healthcare and non-healthcare costs were estimated to be 2.02 billion CAN dollars in 2004. There were 374 deaths attributed to schizophrenia. This combined with the high unemployment rate due to schizophrenia resulted in an additional productivity morbidity and mortality loss estimate of 4.83 billion CAN dollars, for a total cost estimate in 2004 of 6.85 billion CAN dollars. By far the largest component of the total cost estimate was for productivity losses associated with morbidity in schizophrenia (70% of total costs) and the results showed that total cost estimates were most sensitive to alternative assumptions regarding the additional unemployment due to schizophrenia in Canada. CONCLUSIONS: Despite significant improvements in the past decade in pharmacotherapy, programs and services available for patients with schizophrenia, the economic burden of schizophrenia in Canada remains high. The most significant factor affecting the cost of schizophrenia in Canada is lost productivity due to morbidity. Programs targeted at improving patient symptoms and functioning to increase workforce participation has the potential to make a significant contribution in reducing the cost of this severe mental illness in Canada. PMID- 16368054 TI - Effects of rosiglitazone added to submaximal doses of metformin compared with dose escalation of metformin in type 2 diabetes: the EMPIRE Study. AB - OBJECTIVE: This study was designed to compare the efficacy, safety and tolerability of rosiglitazone (RSG) added to submaximal doses of metformin (MET) with dose escalation to the maximal effective dose of MET monotherapy in type 2 diabetes mellitus. RESEARCH DESIGN AND METHODS: In this multi-center, double blind, randomized, parallel-group study, 766 subjects with a baseline MET dose of 1000 mg/day were randomized to receive either RSG 4 mg/day (4 mg/1000 mg) or MET 500 mg/day (1500 mg/day total dose) for 8 weeks. Only the RSG dose was increased in the combination group - to 8 mg/day (8 mg/1000 mg) - and only the MET dose was increased in the MET monotherapy group - to 2000 mg/day for the remaining 16 weeks. RESULTS: After 24 weeks, RSG added to MET (8 mg/1000 mg/day) was at least as effective as 2000 mg/day of MET in improving HbA(1c), with mean reductions of 0.93% (95% CI: -1.06%, -0.80%) and -0.71% (95% CI: -0.83%, -0.60%), respectively, from baseline in subjects that completed the study according to the investigator (mean treatment effect/difference of -0.20% [95% CI: -0.36%, -0.04%]). In addition, a higher percentage of subjects in the RSG + MET group achieved American Diabetes Association target levels of HbA(1c) < 7% (58.1% versus 48.4%) and American Association of Clinical Endocrinologists target levels of HbA(1c) 25 IU/ml). After comparing these ALT and AST abnormal patients with other ALT and AST normal chronic carriers, no statistical difference was observed in the OD values of the anti-HBe (p > 0.05). The ELISA results showed the anti-HBc IgG subclass pattern was IgG1 > IgG3 > IgG4 in chronic carriers and IgG3 > IgG1 > IgG4 in recovered individuals (p < 0.05). This result suggests the IgG1/IgG3 ratio may be related with HBV status. However, in spite of the different anti-HBc IgG1/IgG3 patterns demonstrated in different populations, both anti-HBc IgG1 and IgG3 concentrations were significantly higher in chronic carriers (p < 0.05). Therefore, both the anti-HBc IgG1/IgG3 ratio and their amounts differed. They may play a significant role in chronic carriers and recovered individuals. The anti-HBc IgG subclass profiles of chronic carriers were not changed regardless of liver inflammation, and were independent of sex and age. PMID- 16368068 TI - Are car drivers holding a motorcycle licence less responsible for motorcycle--car crash occurrence? A non-parametric approach. AB - The purpose of this work is to evaluate the effect of a specific motorcycle licence, held by car drivers, in responsibility for motorcycle-car crashes. The data were provided by a multicentric case-control study (MAIDS) regarding the risk of crash and serious injuries of motorcyclists. A non-parametric method, classification and regression tree (CART), was used to accomplish the objective, and then compared to standard unconditional logistic regression. Drivers owning a motorcycle licence turned out to be less responsible for motorcycle-car crashes than drivers who do not have one; both types of analysis are consistent with this result. It is reasonable to assume that car drivers who hold a motorcycle licence have acquired more ability in riding and controlling two wheeled vehicles than drivers without a licence, and this may help them in predicting motorcycles manoeuvres. PMID- 16368069 TI - Factors contributing to officers' fatigue in high-speed maritime craft operations. AB - The purpose of this study was to investigate the difference in the level of fatigue induced by high-speed maritime craft operation between the day- and night shift officers. The demographic and work-related factors that contribute to fatigue were also explored. A total of 93 high-speed maritime craft officers participated in the survey, of whom 35 worked a day shift (Mean age=48.3 years) and 58 worked a night shift (Mean age=45.8 years). Fatigue experience was measured with the Chinese version of the Swedish Occupational Fatigue Inventory (SOFI-C) at the beginning, in the middle and at the end of the work shift. Information on age, work experience, perceived voyage difficulty and duty schedule was obtained. The study found that the night-shift officers seemed to manifest an overall higher level of perceived fatigue than the day-shift officers, while the day-shift officers demonstrated a fatigue carry-over effect across the two workdays. Besides the shift pattern, age, experience in operating high-speed maritime craft and perceived voyage difficulty were the significant factors contributing to the officers' fatigue experience. The finding that the fatigue experience associated with high-speed maritime craft operation has a rapid and accumulative nature suggests that different occupational safety and health guidelines should be devised for these two groups of officers. PMID- 16368070 TI - Photoisomerisable cholesterol derivatives as photo-trigger of liposomes: effect of lipid polarity, temperature, incorporation ratio, and cholesterol. AB - Three cholesterol derivatives containing an azobenzene moiety with different polarities were designed and synthesized (AB lipids 1 to 3). The effects of structure, temperature and incorporation ratio on liposomes were studied, with the results showing that the polarity in 4-substituent and in some cases, 4' substituent may be important for their incorporation feasibility and photoisomerizability in liposomes. Liposomes incorporated with AB lipid 3 could release multi-pulsatilely upon UV and visible light irradiation both in gel state and liquid crystal state of liposomes. An increase in the incorporation ratio of AB lipid 3 enhanced the amount of drug released greatly. Unlike other azobenzene photo-triggers reported, AB lipid 3 did not increase the spontaneous release of liposomes. Furthermore, cholesterol suppressed the spontaneous release of liposomes. PMID- 16368071 TI - Transfer of the AQP1 cDNA for the correction of radiation-induced salivary hypofunction. AB - The treatment of most patients with head and neck cancer includes ionizing radiation (IR). Salivary glands in the IR field suffer significant and irreversible damage, leading to considerable morbidity. Previously, we reported that adenoviral (Ad)-mediated transfer of the human aquaporin-1 (hAQP1) cDNA to rat [C. Delporte, B.C. O'Connell, X. He, H.E. Lancaster, A.C. O'Connell, P. Agre, B.J. Baum, Increased fluid secretion after adenoviral-mediated transfer of the aquaporin-1 cDNA to irradiated rat salivary glands. Proc. Natl. Acad. Sci. U S A. 94 (1997) 3268-3273] and miniature pig [Z. Shan, J. Li, C. Zheng, X. Liu, Z. Fan, C. Zhang, C.M. Goldsmith, R.B. Wellner, B.J Baum, S. Wang. Increased fluid secretion after adenoviral-mediated transfer of the human aquaporin-1 cDNA to irradiated miniature pig parotid glands. Mol. Ther. 11 (2005) 444-451] salivary glands approximately 16 weeks following IR resulted in a dose-dependent increase in salivary flow to > or =80% control levels on day 3. A control Ad vector was without any significant effect on salivary flow. Additionally, after administration of Ad vectors to salivary glands, no significant lasting effects were observed in multiple measured clinical chemistry and hematology values. Taken together, the findings show that localized delivery of AdhAQP1 to IR damaged salivary glands is useful in transiently increasing salivary secretion in both small and large animal models, without significant general adverse events. Based on these results, we are developing a clinical trial to test if the hAQP1 cDNA transfer strategy will be clinically effective in restoring salivary flow in patients with IR-induced parotid hypofunction. PMID- 16368072 TI - A preliminary investigation of acceptance and commitment therapy as a treatment for chronic skin picking. AB - The effectiveness of a deliberately limited version of Acceptance and Commitment Therapy (ACT) for chronic skin picking was evaluated in a pair of multiple baseline across participants designs. Self-monitoring of skin picking showed that four of the five participants reached near zero levels of picking by post treatment, but these gains were not fully maintained for three of the four participants at follow-up. The findings of the self-reported skin picking were generally corroborated by ratings of photographs of the damaged areas and by ratings on a validated measure of skin picking severity. All participants rated the intervention as socially acceptable, and reductions were found on measures of anxiety, depression, and experiential avoidance for most participants as a result of the intervention. Results support the construction of more comprehensive ACT protocols for skin picking. PMID- 16368073 TI - Experienced disgust causes a negative interpretation bias: a causal role for disgust in anxious psychopathology. AB - This paper reports the results of an experiment investigating the effect of induced disgust on interpretational bias using the homophone spelling task. Four groups of participants experienced a disgust, anxiety, happy or neutral mood induction and then completed the homophone spelling task which requires the participant to interpret ambiguous words presented through headphones. Both the disgust and anxiety groups interpreted significantly more threat/neutral homophones as threat than both the happy and neutral groups; the disgust group also interpreted significantly fewer positive/neutral homophones as positive than the happy group. These findings are consistent with the view that induced disgust causes a negative interpretational bias which is similar to that reported for anxiety. The results could not be interpreted in terms of the disgust induction concurrently raising levels of self-reported anxiety, but could be interpreted in terms of disgust maintaining existing levels of anxiety. The effect of disgust was to facilitate negative interpretations rather than emotional interpretations regardless of valence. These findings provide the basis for a causal role for disgust in anxious psychopathology. Because the effect is a non-specific emotion congruent one, elevated disgust levels will result in a predisposition to interpret information in a threatening way across a broad range of anxious- and threat-relevant domains. PMID- 16368074 TI - Hypnotherapy and cognitive behaviour therapy of acute stress disorder: a 3-year follow-up. AB - The long-term benefits of cognitive behaviour therapy (CBT) for trauma survivors with acute stress disorder were investigated by assessing patients 3 years after treatment. Civilian trauma survivors (n=87) were randomly allocated to six sessions of CBT, CBT combined with hypnosis, or supportive counselling (SC), 69 completed treatment, and 53 were assessed 2 years post-treatment for post traumatic stress disorder (PTSD) with the Clinician-Administered PTSD Scale. In terms of treatment completers, 2 CBT patients (10%), 4 CBT/hypnosis patients (22%), and 10 SC patients (63%) met PTSD criteria at 2-years follow-up. Intent-to treat analyses indicated that 12 CBT patients (36%), 14 CBT/hypnosis patients (46%), and 16 SC patients (67%) met PTSD criteria at 2-year follow-up. Patients who received CBT and CBT/hypnosis reported less re-experiencing and less avoidance symptoms than patients who received SC. These findings point to the long-term benefits of early provision of CBT in the initial month after trauma. PMID- 16368075 TI - Neuronal and astrocytic shuttle mechanisms for cytosolic-mitochondrial transfer of reducing equivalents: current evidence and pharmacological tools. AB - The malate-aspartate shuttle and the glycerol phosphate shuttle act to transfer reducing equivalents from NADH in the cytosol to the mitochondria since the inner mitochondrial membrane is impermeable to NADH and NAD+. This transfer of reducing equivalents is essential for maintaining a favorable NAD+/NADH ratio required for the oxidative metabolism of glucose and synthesis of neurotransmitters in brain. There is evidence that both the malate-aspartate shuttle and glycerol phosphate shuttle function in brain; however, there is controversy about the relative importance and cellular localization of these shuttles. The malate-aspartate shuttle is considered the most important shuttle in brain. It is particularly important in neurons and may be extremely low, or even non-existent in brain astrocytes. Several studies provide evidence of glycerol phosphate shuttle activity in brain cells; however, the activity of this shuttle in brain has been questioned. A number of pharmacological tools, including aminooxyacetic acid, beta-methyleneaspartate, phenylsuccinate, and 3-nitropropionic acid, have been used to inhibit the four enzymes and two carrier proteins that participate in the malate-aspartate shuttle. Although no drugs completely inhibit the glycerol phosphate shuttle, evidence for the existence of this shuttle is provided by studies using drugs to inhibit the malate-aspartate shuttle. This report evaluates the evidence for each shuttle in brain cells and the drugs that can be used as pharmacological tools to study these shuttles. PMID- 16368076 TI - A multivariate electrophysiological endophenotype, from a unitary cohort, shows greater research utility than any single feature in the Western Australian family study of schizophrenia. AB - BACKGROUND: Previous studies have found several electrophysiological endophenotypes that each co-varies individually with schizophrenia. This study extends these investigations to compare and contrast four electrophysiological endophenotype, mismatch negativity, P50, P300, and antisaccades, and analyze their covariance on the basis of a single cohort tested with all paradigms. We report a multivariate endophenotype that is maximally associated with diagnosis and evaluate this new endophenotype with respect to its application to genetic analysis. METHODS: Group differences and covariance were analyzed for probands (n = 60), family members (n = 53), and control subjects (n = 44). Associations between individual endophenotypes and diagnostic groups, as well as between the multivariate endophenotype and diagnostic groups, were investigated with logistic regression. RESULTS: Results from all four individual endophenotypes replicated previous findings of deficits in the proband group. The P50 and P300 endophenotypes similarly replicated significant deficits in the family member group, whereas mismatch negativity and antisaccade measures showed a trend. There was minimal correlation between the different endophenotypes. A logistic regression model based on all four features significantly represented the diagnostic grouping (chi(2) = 32.7; p < .001), with 80% accuracy in predicting group membership. CONCLUSIONS: A multivariate endophenotype, based on a weighted combination of electrophysiological features, provides greater diagnostic classification power than any single endophenotype. PMID- 16368077 TI - Glucocorticoids and relapse of major depression (dexamethasone/corticotropin releasing hormone test in relation to relapse of major depression). AB - BACKGROUND: Knowledge of pathogenic mechanisms and predictors of relapse in major depressive disorder is still limited. Hypothalamic-pituitary-adrenocortical (HPA) axis dysregulation is thought to be related to the development and course of depression. METHODS: We investigated whether dexamethasone/corticotropin releasing hormone (DEX/CRH) test parameters were related to the occurrence of relapse in 45 outpatients with clinically remitted major depression. The DEX/CRH test was administered before and after 8 weeks of antidepressant treatment. RESULTS: Posttreatment maximal adrenocorticotropic hormone (ACTH) and maximal cortisol levels, as well as delta ACTH and delta cortisol levels, were significantly higher (all p < .05) among patients who relapsed (n = 22) compared with patients in whom no relapse occurred (n = 23). Higher posttreatment maximal cortisol response on the DEX/CRH test was associated with shorter "relapse-free survival" (p = .05). CONCLUSIONS: In outpatients with clinically remitted major depression, higher posttreatment maximal cortisol levels on the DEX/CRH test were associated with relapse of major depression. PMID- 16368078 TI - Size of treatment effects and their importance to clinical research and practice. AB - In randomized clinical trails (RCTs), effect sizes seen in earlier studies guide both the choice of the effect size that sets the appropriate threshold of clinical significance and the rationale to believe that the true effect size is above that threshold worth pursuing in an RCT. That threshold is used to determine the necessary sample size for the proposed RCT. Once the RCT is done, the data generated are used to estimate the true effect size and its confidence interval. Clinical significance is assessed by comparing the true effect size to the threshold effect size. In subsequent meta-analysis, this effect size is combined with others, ultimately to determine whether treatment (T) is clinically significantly better than control (C). Thus, effect sizes play an important role both in designing RCTs and in interpreting their results; but specifically which effect size? We review the principles of statistical significance, power, and meta-analysis, and commonly used effect sizes. The commonly used effect sizes are limited in conveying clinical significance. We recommend three equivalent effect sizes: number needed to treat, area under the receiver operating characteristic curve comparing T and C responses, and success rate difference, chosen specifically to convey clinical significance. PMID- 16368079 TI - GABA(B) receptor function and subunit expression in the rat spinal cord as indicators of stress and the antinociceptive response to antidepressants. AB - Experiments were undertaken to examine whether once daily i.p. administration of either of two antidepressants used for the treatment of neuropathic pain, amitriptyline (10 mg/kg) and fluoxetine (5 mg/kg), to rats for 7 days modifies GABA(B) receptor function and subunit expression in the lumbar spinal cord. The results indicate that, as previously reported for desipramine, both amitriptyline and fluoxetine increase the pain threshold to a thermal stimulus, the expression of GABA(B(1)) subunits, and baclofen-stimulated [35S]GTPgammaS binding, a measure of GABA(B) receptor function. The effects of antidepressant administration on GABA(B(1b)) and GABA(B(2)) subunit expression in spinal cord are more variable than for GABA(B(1a)). It was also discovered that repeated daily exposure to a thermal stimulus or immobilization stress increases GABA(B(1a)) expression in the lumbar spinal cord, with no commensurate change in thermal pain threshold or GABA(B) receptor sensitivity. These results support a relationship between GABA(B) receptors and the action of antidepressants. The findings demonstrate that drug-induced increases in GABA(B) receptor function can occur independently of any change in GABA(B) receptor subunit expression and are consistent with the notion that GABA(B) receptor subunits have multiple functions, only one of which is dimerization to form GABA(B) receptors. The data also suggest that GABA(B) subunit gene expression may serve as a preclinical marker of antidepressant efficacy and of drug- or stress-induced modifications in central nervous system activity. PMID- 16368080 TI - Calbindin and calretinin immunoreactivity in the retina of adult and larval sea lamprey. AB - The presence of calretinin and calbindin immunoreactivity is studied in the retina of larval and adult lamprey and their respective distributions are compared. Calretinin distribution is also studied in the retina of transforming stages. Western blot analysis in brain extracts showed a 29-kDa band with both polyclonal anti-calbindin and anti-calretinin antibodies. Calbindin and calretinin immunoreactivity has shown a partially different distribution. In the adult retina large and small bipolar cells, with respectively stratified or diffuse axons, the inner row of horizontal cells and ganglion cells and/or some amacrine cells were labeled with anti-calretinin antibody. The anti-calbindin antibody labels the same cell types except most of ganglion cells, but the label was less conspicuous. Therefore, the possible existence of these two calcium binding proteins in the central nervous system of the sea lamprey could be discussed. In the differentiated central retina of larval lampreys, numerous calretinin immunoreactive bipolar and ganglion cells were observed, while, in the lateral retina, only ganglion cells were labeled, accordingly with the lack of differentiation of other neural cell types. CR-ir bipolar cells appeared in the retina by the stage 5 of transformation, i.e. about the time when differentiation of photoreceptors occurs. The comparison of the distribution of calretinin and calbindin between adult and larval central retina of lampreys shows striking differences that could be related to the different functionality of eyes in these two stages of the life cycle of lampreys. In addition, this is the first report on the presence of calcium-binding proteins in the larval and transforming lamprey retina, on the presence of calretinin- and calbindin-immunoreactive horizontal cells in adult lamprey retinas and on the differential stratification of bipolar cell terminals. PMID- 16368081 TI - Exogenous erythropoietin provides neuroprotection of grafted dopamine neurons in a rodent model of Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disease marked by severe loss of dopamine (DA) neurons in the nigrostriatal system, which results in depletion of striatal DA. Transplantation of embryonic ventral mesencephalic (VM) DA neurons into the striatum is a currently explored experimental treatment aimed at replacing lost DA in the nigrostriatal system, but is plagued with poor survival (5-20%) of implanted neurons. Here, we tested the ability of erythropoietin (Epo) to provide neuroprotection for embryonic day 14 (E14) VM DA neurons. Epo was tested in vitro for the ability to augment tyrosine hydroxylase-immunoreactive (TH-ir) neuron survival under normal cell culture conditions. In vitro, Epo did not increase the number of TH-ir neurons when administered at the time of plating the E14 VM cells in culture. We also tested the efficacy of Epo to enhance E14 VM transplants in vivo. Rats unilaterally lesioned with 6-hydroxydopamine received transplants that were incubated in Epo. Treatment with Epo produced significant increases in TH-ir neuron number, soma size, and staining intensity. Animals receiving Epo-treated grafts exhibited significantly accelerated functional improvements and significantly greater overall improvements from rotational asymmetry compared to control grafted rats. These data indicate that the survival of embryonic mesencephalic TH-ir neurons is increased when Epo is administered with grafted cells in a rodent model of PD. As direct neurotrophic effects of Epo were not observed in vitro, the mechanism of Epo neuroprotection remains to be elucidated. PMID- 16368082 TI - Modulation of early auditory processing by visually based sound prediction. AB - Brain activity was measured by magnetoencephalography (MEG) to investigate whether the early auditory system can detect changes in audio-visual patterns when the visual part is presented earlier. We hypothesized that a template underlying the mismatch field (MMF) phenomenon, which is usually formed by past sound regularities, is also used in visually based sound prediction. Activity similar to the MMF may be elicited by comparing an incoming sound with the template. The stimulus was modeled after a keyboard: an animation in which one of two keys was depressed was accompanied by either a lower or higher tone. Congruent audio-visual pairs were designed to be frequent and incongruent pairs to be infrequent. Subjects were instructed to predict an incoming sound based on key movement in two sets of trials (prediction condition), whereas they were instructed not to do so in the other two sets (non-prediction condition). For each condition, the movement took 50 ms in one set (Delta = 50 ms) and 300 ms in the other (Delta = 300 ms) to reach the bottom, at which time a tone was delivered. As a result, only under the prediction condition with Delta = 300 ms was additional activity for incongruent pairs observed bilaterally in the supratemporal area within 100-200 ms of the auditory stimulus onset; this activity had spatio-temporal properties similar to those of MMF. We concluded that a template is created by the visually based sound prediction only after the visual discriminative and sound prediction processes have already been performed. PMID- 16368087 TI - Bone marrow-derived myocyte-like cells and regulation of repair-related cytokines after bone marrow cell transplantation. AB - OBJECTIVE: Whether bone marrow cells injected following acute myocardial infarction (MI) transdifferentiate into cardiomyocytes remains controversial, and how these cells affect repair-related cytokines is not known. METHODS: Autologous bone marrow-derived mononuclear cells (BM-MNCs) labeled with DiI, 1,1' dioctadecyl-1 to 3,3,3',3'-tetramethylindocarbocyanine perchlorate, or saline were intravenously injected into rabbits 5 h following a 30-min ischemia and reperfusion protocol, and cardiac function and the general pathology of the infarcted heart were followed up 1 and 3 months post-MI. To search for regenerated myocardium, electron microscopy as well as confocal microscopy were performed in the infarcted myocardium 7 days post-MI. Expression levels of repair related cytokines were evaluated by immunohistochemistry and Western blotting. RESULTS: Improvements in cardiac function and reductions in infarct size were observed in the BM-MNC group 1 month and 3 months post-MI. Using electron microscopy 7 days after infarction, clusters of very immature (fetal) and relatively mature cardiomyocytes undergoing differentiation were identified in the infarcted anterior LV wall in the BM-MNC group, though their numbers were small. These cells contained many small and dense DiI particles (a BM-MNC marker), indicating that cardiomyocytes had regenerated from the injected BM MNCs. The expression of both transforming growth factor-beta, which stimulates collagen synthesis and matrix metalloproteinase-1, a collagenase, were both down regulated 7 days and 1 month post-MI in the BM-MNC group. Stromal cell-derived factor-1, which is known to recruit BM-MNCs into target tissues, was overexpressed in the infarcted areas of BM-MNC hearts 7 days post-MI. CONCLUSIONS: Intravenous transplantation of BM-MNCs leads to the development of BM-MNC-derived myocyte-like cells and regulates the expression of repair-related cytokines that facilitate repair following myocardial infarction. PMID- 16368088 TI - GSH depletion after erythrocytes acidic treatment is related to intracellular hemoglobin levels. PMID- 16368089 TI - Preferential posterior cerebellum defect in BETA2/NeuroD1 knockout mice is the result of differential expression of BETA2/NeuroD1 along anterior-posterior axis. AB - BETA2/NeuroD1 has been shown to play a major role in terminal differentiation of the pancreatic and enteroendocrine cells, as well as for the survival of photoreceptors. Here, we report that the loss of BETA2/NeuroD1 affected the cerebellar development with a major reduction of granule cell number. However, there is a differential reduction of granule cells along the anterior and posterior axis of the cerebellum; while the reduction of granule cells in the anterior lobes is substantial, there is an almost complete loss of granule cells in the posterior compartment. To understand the mechanism for this anterior posterior difference, we carried out detailed analyses. We found that both BETA2/NeuroD1 and its direct target TrkC, expression commence earlier in the posterior part than those in the anterior part during cerebellum development. Consequently, loss of BETA2/NeuroD1 enhances granule cell death in the posterior 2 days earlier than the anterior. Furthermore, the higher rate of cell death in the posterior of the cerebellum is concomitant with the reduction of TrkC expression in knockout mice. Thus, our data indicate that preferential expression of BETA2/NeuroD1 and TrkC in posterior lobes explains the earlier start of cell apoptosis and preferential loss of granule cells in the posterior lobes. PMID- 16368090 TI - Pharmacological characterisation of sodium channels in sinoatrial node pacemaking in the rat heart. AB - Blockade of sodium channels located in the sinoatrial node can slow diastolic depolarisation rate, recorded in vitro. The objective was therefore to determine whether these blockers could slow heart rate in vivo. The heart rate was firstly measured in spontaneously beating, isolated rat heart atria in the presence of different voltage gated sodium channel blockers. Tetrodotoxin and lidocaine slightly reduced heart rate whereas KC 12291 and R 56865, which mainly interact with the persistent component of the sodium current, concentration dependently and potently induced bradycardia. In the pithed rat, tetrodotoxin induced statistically significant decreases heart rate, maximal effects were: -32.2+/-6.1 beat per min. KC 12291 and R 56865 dose-dependently induced bradycardia (Delta heart rate obtained, -55.1+/-5.2 beat per min, P<0.05, and -71.9+/-8.5 bpm, P<0.05, respectively). In conclusion, voltage gated sodium channel blockers rather selective for the persistent current, exert a potent bradycardic effect in the rat. PMID- 16368091 TI - Involvement of Egr-1 in lens epithelial cell death induced by selenite. AB - Selenite-overdose cataract in young rats may be caused by an initial insult to the lens epithelial cells. Our previous DNA array analysis revealed a significant increase in the expression of mRNA for early growth response protein-1 (Egr-1) in lens epithelial cells after injection of selenite. This suggested that up regulation of Egr-1 mRNA may be involved in lens epithelial cell death. The purpose of the present experiment was to further clarify the involvement of Egr-1 in lens epithelial cell death induced by selenite. Rat lens epithelial explants were cultured with sodium selenite. Selenite caused epithelial explants to leak LDH into the medium. During LDH leakage, increased expression of mRNA for Egr-1 was observed by RT-PCR. To further test the involvement of Egr-1 in selenite induced cell death, mouse lens epithelial cell line (alpha-TN4 cells) was treated with antisense oligonucleotide for Egr-1. Antisense oligonucleotide for Egr-1 significantly diminished expression of Egr-1 protein and leakage of LDH. These results suggested that increased activity of Egr-1 may be a factor in lens epithelial cell death induced by selenite. PMID- 16368092 TI - Toxoplasma gondii: DNA vaccination with bradyzoite antigens induces protective immunity in mice against oral infection with parasite cysts. AB - Infection of the host by Toxoplasma gondii leads to an acute systemic dissemination of tachyzoites, followed by a chronic phase, in which bradyzoites, enclosed in cysts, persist in the brain, the heart, and other tissues. Among putative vaccine candidates, the bradyzoite antigens BAG1 and MAG1 look promising since they are preferentially expressed during the chronic stage of the parasite. This work focused on studying the immunogenicity of bradyzoite antigens in a mouse model of chronic toxoplasmosis. A mixture of plasmids directing the cytoplasmic expression of MAG1 and BAG1 in mammalian cells was used to immunize mice. We show here that immunized mice developed, preferentially, specific anti MAG1 and anti-BAG1 IgG2a subclass antibodies, indicating a shift towards a Th1 like response after DNA immunization. We then demonstrated that DNA immunization followed by challenge infection elicited effective protection in mice, suggesting that bradyzoite antigens should be considered in the design of vaccines against toxoplasmosis. PMID- 16368093 TI - Constitutive activity of a UV cone opsin. AB - Vertebrate visual pigment proteins contain a conserved carboxylic acid residue in the third transmembrane helix. In rhodopsin, Glu113 serves as a counterion to the positively charged protonated Schiff base formed by 11-cis retinal attached to Lys296. Activation involves breaking of this ion pair. In UV cone pigments, the retinyl Schiff base is unprotonated, and hence such a salt bridge is not present; yet the pigment is inactive in the dark. Mutation of Glu108, which corresponds to rhodopsin's Glu113, to Gln yields a pigment that remains inactive in the dark. The apoproteins of both the wild-type and mutant, however, are constitutively active with the mutant being of significantly higher activity. Thus, one important role for preserving the negatively charged glutamate in the third helix of UV pigments is to maintain a less active opsin in a manner similar to rhodopsin. Ligand binding itself in the absence of a salt bridge is sufficient for deactivation. PMID- 16368094 TI - Development and efficiency of an acute knee trauma list. AB - Following acute knee trauma some patients will require urgent arthroscopy. Traditionally surgery is undertaken as in-patient on the general trauma list. This system leads to blockage of beds, repeated starving of the patient and out of-hour's surgery in many cases. We introduced a dedicated Day Surgery knee trauma list (KTL) to reduce in-patient admissions, waiting time and cost. The typical indications for urgent knee arthroscopy are locked knees secondary to meniscal tears, loose bodies or cruciate ligament ruptures; acute osteochondral fractures; and children with acute meniscal tears. This retrospective review compares the study group: patients on the knee trauma list during the first 8 months following introduction, and the control group: patients on the general in patient trauma list over a similar period immediately prior to implementation of the new list. There were 49 patients in the control group with an average stay in hospital of 2.5 days. Out-of-hour's operations were performed in 13 patients and 3 patients required a repeat arthroscopy. Fifty-three patients were treated in the knee study group. The significant operations were meniscal repair in 7, fixation of osteochondral fragments in 3. None of the patients required overnight stay. Control group patients were more likely to have surgery performed by inexperienced non-specialist knee surgeons, whereas specialist knee surgeons staffed the knee trauma list. Specialist surgeons were available to staff the knee trauma list. As the majority of the control group had spent an average of 2.2 nights in hospital, there has been considerable savings to the hospital. Introduction of the Day Surgical Knee trauma list has reduced unnecessary admissions per week; episodes of prolonged starving, cost and have improved patient satisfaction and management. PMID- 16368095 TI - Facilitating damage control surgery with a dedicated DCS equipment trolley. PMID- 16368096 TI - The specialist palliative care nurse: a qualitative study of the patients' perspective. AB - BACKGROUND: Relatively little is known about patients' perceptions of the work and role of the specialist palliative care nurse. Understanding the patients' perspective can help to evaluate services, improve quality of care, and identify misunderstandings. OBJECTIVES: To explore the experiences of those who said that they had a 'terminal illness', focusing on patients' perceptions of the work and role of these nurses. DESIGN: Qualitative study with narrative interviews. SETTING: England and Wales. PARTICIPANTS: Forty-one people recruited through those working in hospices, community nurses, general practitioners, support groups, a national newspaper, and a conference on palliative care. Twenty-five people talked about the work of specialist palliative care nurses. METHOD: Interviews were fully transcribed; followed by a thematic analysis with constant comparison. RESULTS: Patients valued the nurses' work, particularly their advice on practical matters, information given about their disease, emotional support, advice on symptoms, and help with communication. They were glad that help was readily available. However, some patients who had been referred to the service did not realise that specialist palliative care nurses may be involved at a relatively early stage in a person's illness, and sometimes felt distressed by an early referral. One woman felt she had not had the emotional support she needed and another knew of women who had been upset because these nurses had discussed topics such as place of death 'too early'. However, people recognised the difficulties nurses faced in their work. CONCLUSIONS: Although our study differs from other studies, particularly in the way people were recruited, our findings support previous studies that have shown that specialist palliative care nurses are highly valued by those who have a terminal illness. It is important for people to understand that these nurses may be involved from the time of diagnosis and that roles have changed. PMID- 16368097 TI - Characterization of LST-R533: uncovering a novel repetitive element in Leishmania. AB - We have previously isolated and sequenced a novel repetitive element, now named LST-R533, which is present in four different regions of one extremity of Leishmania major chromosome 20. The repeats are polymorphic in size, ranging from 367 to 533 bp and contain an internal 81 bp sequence with highly conserved segments (14-81 bp long) dispersed throughout the parasite's genome. These sequences were not found in coding regions of any predicted gene in L. major Friedlin genome, but are part of untranslated regions of some Leishmania transcripts. Analysis of the 81 bp sequence revealed significant degrees of identity with retrotransposons described in several other organisms. The presence of the sequence in other species from genus Leishmania was determined by Southern hybridisation and DNA sequencing. This analysis indicated the conservation of the 81-nucleotide element in all the Leishmania species evaluated. No sequences corresponding to LST-R533 or the 81 bp element were found on either Trypanosoma brucei or Trypanosoma cruzi databanks. PMID- 16368098 TI - EnP1 and EnP2, two proteins associated with the Encephalitozoon cuniculi endospore, the chitin-rich inner layer of the microsporidian spore wall. AB - Microsporidia are obligate intracellular parasites forming environmentally resistant spores that harbour a rigid cell wall. This wall comprises an outer layer or exospore and a chitin-rich inner layer or endospore. So far, only a chitin deacetylase-like protein has been shown to localize to the Encephalitozoon cuniculi endospore and either one or two proteins have been clearly assigned to the exospore in two Encephalitozoon species: SWP1 in E. cuniculi, SWP1 and SWP2 in Encephalitozoon intestinalis. Here, we report the identification of two new spore wall proteins in E. cuniculi, EnP1 and EnP2, the genes of which are both located on chromosome I (ECU01_0820 and ECU01_1270, respectively) and have no known homologue. Detected by immunoscreening of an E. cuniculi cDNA library, enp1 is characterized by small-sized 5' and 3' untranslated regions and is highly expressed throughout the whole intracellular cycle. The encoded basic 40 kDa antigen displays a high proportion of cysteine residues, arguing for a significant role of disulfide bridges in spore wall assembly. EnP2 is a 22 kDa serine-rich protein that is predicted to be O-glycosylated and glycosylated phosphatidyl inositol-anchored. Although having been identified by mass spectrometry of a dithiothreitol-soluble fraction, this protein contains only two cysteine residues. Mouse polyclonal antibodies were raised against EnP1 and EnP2 recombinant proteins produced in Escherichia coli Our immunolocalisation data indicate that EnP1 and EnP2 are targeted to the cell surface as early as the onset of sporogony and are finally associated with the chitin-rich layer of the wall in mature spores. PMID- 16368099 TI - Increased atherosclerotic lesion area in apoE deficient mice overexpressing bovine growth hormone. AB - Human growth hormone (GH) excess is linked to increased cardiovascular morbidity and mortality. However, little is known about the effect of GH excess on atherosclerosis. We developed a new mouse model to assess the hypothesis that GH overexpression accelerates atherosclerotic lesion formation. apoE(-/-) mice were crossed with bovine GH (bGH) transgenic mice to yield apoE(-/-) mice overexpressing bGH (apoE(-/-)/bGH). The mice were fed either standard or Western diet. At 22 weeks, atherosclerotic lesion area of thoracic aorta was larger in apoE(-/-)/bGH mice compared with littermate apoE(-/-) mice fed either diet (standard: +161+/-50%, Western: +430+/-134%). Aortic sinus lesions were more severe in apoE(-/-)/bGH mice fed standard diet compared with littermate apoE(-/-) mice. apoE(-/-)/bGH mice had lower (VLDL+LDL)/HDL ratios compared with littermate apoE(-/-) mice, while systolic blood pressure was higher in apoE(-/-)/bGH mice, irrespective of diet. The levels of serum amyloid A and hepatic CRP mRNA were higher in apoE(-/-)/bGH mice than in littermate apoE(-/-) mice. In conclusion, this study shows that excess GH augments the development of atherosclerosis in apoE(-/-) mice. The mechanisms could be direct effects of GH on cellular processes in the vessel wall or the result of concomitant processes such as hypertension or a general inflammatory state. PMID- 16368100 TI - An analytical study on the thermal effects of cryosurgery on selective cell destruction. AB - The aim of cryosurgery is to kill cells within a closely defined region maintained at a predetermined low temperature. To effectively kill cells, it is important to be able to predict and control the cooling rate over some critical range of temperatures and freezing states in order to regulate the spatial extent of injury during any freeze-thaw protocol. The objective of manipulating the freezing parameters is to maximize the destruction of cancer cells within a defined spatial domain while minimizing cryoinjury to the surrounding healthy tissue. An analytical model has been developed to study the rate of cell destruction within a liver tumor undergoing a freeze-thaw cryosurgical process. Temperature transients in the tumor undergoing cryosurgery have been quantitatively investigated. The simulation is based on solving the transient bioheat equation using the finite volume scheme for a single or multiple-probe geometry. Simulated results show good agreement with experimental data obtained from in vivo clinical study. The calibrated model has been employed to study the effects of different freezing rates, freeze-thaw cycle(s), and multi-probe freezing on cell damage in a liver tumor. The effectiveness of each treatment protocol is estimated by generating the cell survival-volume signature and comparing the percentage of cell damaged within the ice-ball. Results from the model show that employing freeze-thaw cycles has the potential to enhance cell destruction within the cancerous tissue. Results from this study provide the basis for designing an optimized cryosurgical protocol which incorporates thermal effects and the extent of cell destruction within tumors. PMID- 16368101 TI - An improved method for sn-2 position analysis of triacylglycerols in edible oils and fats based on immobilised lipase D (Rhizopus delemar). AB - The use of lipase D (Rhizopus delemar) immobilised on microporous polypropylene as a replacement for the standard pancreatic lipases used in the stereospecific sn-2 position analysis of triacylglycerols from edible oils and fats is studied. Excellent hydrolysis characteristics are obtained in hexane/methanol solvents at reaction temperatures up to 60 degrees C with hydrolysis times of only 10-20 min. The favourable conditions for the hydrolysis reaction allow fats with higher melting points to be analysed and facilitate coupling of the hydrolysis reaction to the later steps in the analytical protocol. The performance of the new method is compared to that of the standard method using pancreatic lipase. The novel procedure is faster, manual sample handling is reduced, while the results obtained with both methods are comparable. The influence of alkyl-chain length on hydrolysis rates seems to be negligible for the most common vegetable fatty acids. Acyl migration was found to be absent. The short-term repeatability of the method ranges from 10% for fatty acids present at levels close to the detection limits to less than 1% for the major fatty acids. The detection limit is approximately 0.05%. Although the application of the immobilised enzyme in fully automated sn-2 position analysis seems to be promising, the attempts to do this using a packed bed reactor were not successful due to a rapid loss of enzyme activity. PMID- 16368102 TI - Development of a pressurized liquid extraction and clean-up procedure for the determination of alpha-endosulfan, beta-endosulfan and endosulfan sulfate in aged contaminated Ethiopian soils. AB - Pressurized liquid extraction (PLE) was investigated for the extraction of two endosulfan isomers and their metabolite from two real contaminated soil samples. PLE for 3x10min at 100 degrees C was proven to be more exhaustive than Soxhlet extraction (SOX) in one soil sample. On the other soil sample investigated the method was found to be equally exhaustive as SOX. The use of hazardous organic solvents such as n-hexane, toluene, and diethyl ether has been avoided in PLE and clean-up. Instead less toxic solvents have been used both at the extraction step (acetone/n-heptane) and clean-up step (ethyl acetate/n-heptane). A column Florisil clean-up procedure that consumes relatively low solvent volumes has been optimized and applied to purify soil extracts. The developed analytical procedure was validated by applying it to a certified reference soil material (CRM811-050). A recovery of 103% total endosulfan residue was obtained versus certified values. PMID- 16368103 TI - Interpreting the effects of temperature and solvent composition on separation of amino-acid racemates by chiral ligand-exchange chromatography. AB - Routinely applied at both preparative and analytical scales, chiral ligand exchange chromatography (CLEC) separates enantiomers capable of chelating a divalent transition-metal-ion through a pair of coordinating electronegative atoms. CLEC separation efficiencies are strongly dependent on column operating conditions, including temperature and mobile-phase solvent composition. Although previous empirical studies provide some useful guidelines for optimizing column operating conditions, the fundamental mechanisms underlying the unusually high sensitivity of CLEC performance to operating temperature and solvent composition remain poorly understood, limiting efforts to develop a comprehensive model for the technology. To address this problem, we report transport and chemical equilibria data for the separation of alpha-amino acids on a Nucleosil chiral-1 column presenting L-hydroxyproline as the immobilized ligand. Solute transport is found to be limited by pore diffusion at all column operating temperatures and solvent compositions, validating the existence of local equilibria throughout the column. Changes in separation performance are found to correlate with changes in chemical equilibria, emphasizing the need to carefully account for all speciation within the column when modeling CLEC and providing important fundamental data to achieve this goal. Each enantiomer participates in a large number of solution phase complexes. As a result, the thermodynamic driving force for separation is unusually complex, allowing subtle changes in column operating conditions to mediate significant changes in speciation profiles and separation efficiency. A reaction-equilibria model accounting for all speciation within the CLEC column is proposed and used to estimate enantiomer partition coefficients and retention times. PMID- 16368104 TI - Supermacroporous cryogel matrix for integrated protein isolation. Immobilized metal affinity chromatographic purification of urokinase from cell culture broth of a human kidney cell line. AB - A new type of supermacroporous, monolithic, cryogel affinity adsorbent was developed, allowing the specific capture of urokinase from conditioned media of human fibrosarcoma cell line HT1080. The affinity adsorbent was designed with the objective of using it as a capture column in an integrated perfusion/protein separation bioreactor setup. A comparative study between the utility of this novel cryogel based matrix and the conventional Sepharose based affinity matrix for the continuous capture of urokinase in an integrated bioreactor system was performed. Cu(II)-ion was coupled to epoxy activated polyacrylamide cryogel and Sepharose using iminodiacetic acid (IDA) as the chelating ligand. About 27-fold purification of urokinase from the conditioned culture media was achieved with Cu(II)-IDA-polyacrylamide cryogel column giving specific activity of about 814 Plough units (PU)/mg protein and enzyme yields of about 80%. High yields (95%) were obtained with Cu(II)-IDA-Sepharose column by virtue of its high binding capacity. However, the adsorbent showed lower selectivity as compared to cryogel matrix giving specific activity of 161 PU/mg protein and purification factor of 5.3. The high porosity, selectivity and reasonably good binding capacity of Cu(II)-IDA-polyacrylamide cryogel column make it a promising option for use as a protein capture column in integrated perfusion/separation processes. The urokinase peak pool from Cu(II)-IDA-polyacrylamide cryogel column could be further resolved into separate fractions for high and low molecular weight forms of urokinase by gel filtration chromatography on Sephacryl S-200. The selectivity of the cryogel based IMAC matrix for urokinase was found to be higher as compared to that of Cu(II)-IDA-Sepharose column. PMID- 16368105 TI - Geometric scaling effects on instrumental plate height in field flow fractionation. AB - This paper examines geometric scaling models for field flow fractionation systems to understand how channel dimensions affect resolution and retention. Specifically, the changing contribution of the instrumental plate height during miniaturization of field flow fractionation (FFF) systems is reported. The work is directed towards determining the optimal geometrical parameters for miniaturization of field flow fractionation systems. The experimental relationship between channel height in FFF systems and instrumental plate heights is reported. FFF scaling models are modified to: (i) better clarify the dependence of plate height and resolution on channel height in FFF and (ii) include a more complete geometrical scaling analysis and model comparison in the low retention regime. Electrical field flow fractionation has been shown to benefit from miniaturization, so this paper focuses on that subtype, but surprisingly, the results also indicate the possibility of improvement in performance with miniaturization of other field flow fractionation systems including general FFF subtypes in which the applied field does not vary with channel height. This paper also discusses the potential role of more powerful microscale field flow fractionation systems as a new class of sample preparation units for micro-total-analysis systems (mu-TAS). PMID- 16368106 TI - Follicular atresia during Dacus oleae oogenesis. AB - Programmed cell death, constitutes a common fundamental incident that occurs during oogenesis in a variety of different animals. It plays a significant role in the maturation process of the female gamete and also in the removal of abnormal and superfluous cells at certain checkpoints of development. In the present study, we demonstrate the existence of follicular atresia during mid oogenesis in the olive fruit fly Dacus oleae (Tephritidae). The number of atretic follicles increases following the age of the fly, suggesting for the presence of an age-susceptible process. The atretic follicles contain nurse cells that exhibit chromatin condensation, DNA fragmentation and actin cytoskeleton alterations, as revealed by propidium iodide staining, TUNEL labeling and phalloidin-FITC staining. Conventional light and electron microscopy disclose that the nurse cell remnants are phagocytosed by the adjacent follicle cells. The follicular epithelium also eliminates the oocyte through phagocytosis, resulting to an egg chamber with no compartmentalized organization. The data presented herein are very similar compared to previous reported results in other Diptera species, strongly suggesting the occurrence of a phylogenetically conserved mechanism of follicular atresia. All these observations also support the notion that mid-oogenesis in D. oleae may be the critical regulation point at which superfluous and defective egg chambers are selectively eliminated before they reach maturity. PMID- 16368107 TI - BCL-XL dimerization by three-dimensional domain swapping. AB - Dimeric interactions among anti- and pro-apoptotic members of the BCL-2 protein family are dynamically regulated and intimately involved in survival and death functions. We report the structure of a BCL-X(L) homodimers a 3D-domain swapped dimer (3DDS). The X-ray crystal structure demonstrates the mutual exchange of carboxy-terminal regions including BH2 (Bcl-2 homology 2) between monomer subunits, with the hinge region occurring at the hairpin turn between the fifth and sixth alpha helices. Both BH3 peptide-binding hydrophobic grooves are unoccupied in the 3DDS dimer and available for BH3 peptide binding, as confirmed by sedimentation velocity analysis. BCL-X(L) 3DDS dimers have increased pore forming activity compared to monomers, suggesting that 3DDS dimers may act as intermediates in membrane pore formation. Chemical crosslinking studies of Cys substituted BCL-X(L) proteins demonstrate that 3DDS dimers form in synthetic lipid vesicles. PMID- 16368108 TI - Structural prediction of peptides bound to MHC class I. AB - An ab initio structure prediction approach adapted to the peptide-major histocompatibility complex (MHC) class I system is presented. Based on structure comparisons of a large set of peptide-MHC class I complexes, a molecular dynamics protocol is proposed using simulated annealing (SA) cycles to sample the conformational space of the peptide in its fixed MHC environment. A set of 14 peptide-human leukocyte antigen (HLA) A0201 and 27 peptide-non-HLA A0201 complexes for which X-ray structures are available is used to test the accuracy of the prediction method. For each complex, 1000 peptide conformers are obtained from the SA sampling. A graph theory clustering algorithm based on heavy atom root-mean-square deviation (RMSD) values is applied to the sampled conformers. The clusters are ranked using cluster size, mean effective or conformational free energies, with solvation free energies computed using Generalized Born MV 2 (GB MV2) and Poisson-Boltzmann (PB) continuum models. The final conformation is chosen as the center of the best-ranked cluster. With conformational free energies, the overall prediction success is 83% using a 1.00 Angstroms crystal RMSD criterion for main-chain atoms, and 76% using a 1.50 Angstroms RMSD criterion for heavy atoms. The prediction success is even higher for the set of 14 peptide-HLA A0201 complexes: 100% of the peptides have main-chain RMSD values < or =1.00 Angstroms and 93% of the peptides have heavy atom RMSD values < or =1.50 Angstroms. This structure prediction method can be applied to complexes of natural or modified antigenic peptides in their MHC environment with the aim to perform rational structure-based optimizations of tumor vaccines. PMID- 16368109 TI - Dynamics and mechanism of the Tanford transition of bovine beta-lactoglobulin studied using heteronuclear NMR spectroscopy. AB - The Tanford transition is a conformational change of bovine beta-lactoglobulin (betaLG) occurring at around pH 7, identified originally on the basis of optical rotatory dispersion and the accessibility of a thiol group. X-ray analysis has suggested that a conformational change to the EF-loop is responsible for the Tanford transition, with the loop closing the hydrophobic cavity of the beta barrel of the betaLG molecule below pH 7 and flipping to open the cavity above pH 7. To clarify the dynamics of this conformational change, NMR measurements were made at neutral pH. Since severe signal broadening due to monomer-dimer equilibrium prevented NMR measurements of wild-type betaLG at neutral pH, we searched for optimal sample conditions, finding that a disulfide bond-linked dimer of the mutant A34C gives an HSQC spectrum without signal broadening. The HSQC and CD spectra indicated that in overall conformation A34C is similar to wild-type betaLG, suggesting that the A34C dimer is a good model with which to study the structure and dynamics of the wild-type at neutral pH. The pH-dependent HSQC signal changes and Lipari-Szabo type relaxation analyses of the A34C dimer revealed that the conformational change to the EF-loop occurs above pH 7. We observed two types of motions in the EF-loop region; relatively fast (micro- to milliseconds) and slow (milliseconds or slower) conformational exchanges of the residues located in the hinge and top of the EF-loop regions, respectively. Furthermore, the GH-loop adjacent to the EF-loop exhibited conformational change at a pH slightly lower than that at which the EF-loop motions occurred. From these observations, we propose a three-step mechanism of conformational change in the EF-loop leading to the Tanford transition, in which the GH-loop conformational change, the cleavage of the hydrogen bonds at the hinge, and the flip of the EF-loop occur sequentially. PMID- 16368110 TI - Dynamics of allostery in hemoglobin: roles of the penultimate tyrosine H bonds. AB - The tyrosine residues adjacent to the C termini of the hemoglobin (Hb) subunits, alphaY140 and betaY145, are expected to play important structural roles, because the C termini are the loci of T-state quaternary salt-bridges, and because the tyrosine side-chains bridge the H and F helices via H bonds to the alphaV93 and betaV98 carbonyl groups. These roles have been investigated via measurements of oxygen binding, (1)H NMR spectra, resonance Raman (RR) spectra, and time-resolved resonance Raman (TR(3)) spectra on site mutants in which the Hcdots, three dots, centeredF H bonds are eliminated by replacing the tyrosine residues with phenylalanine. The TR(3) spectra confirm the hypothesis, based on TR(3) studies of wild-type Hb, that the Hcdots, three dots, centeredF H bonds break and then re form during the sub-microsecond phase of the R-T quaternary transition. The TR(3) spectra support the inference from other mutational studies that the alphabeta dimers act as single dynamic units in this early phase, motions of the E and F helices being coupled tightly across the dimer interface. Formation of T quaternary contacts occurs at about the same rate in the mutants as in HbA. However, these contacts are weakened substantially by the Y/F substitutions. Equilibrium perturbations are apparent also, especially for the alpha-subunits, in which relaxation of the Fe-His bond, strengthening of the Acdots, three dots, centeredE interhelical H bond, and weakening of the "switch" quaternary contact in deoxyHb are all apparent. Structural effects are less marked for the beta chain Y/F replacement, but the Bohr effect is reduced by 25%, indicating that the salt-bridge and H bond interactions of the adjacent C terminus are loosened. The alpha-chain replacement reduces the Bohr effect much more, consistent with the global perturbations detected by the structure probes. PMID- 16368111 TI - The dopamine transporter gene and the impulsivity phenotype in attention deficit hyperactivity disorder: a case-control association study in a Korean sample. AB - The dopamine transporter gene (DAT1) has been extensively studied as one of the candidate genes in attention-deficit/hyperactivity disorder (ADHD). Several studies have reported on the association between the DAT1 10-repeat allele and cognitive variables in ADHD. However, few studies have been designed to ascertain the association between DAT1 genotypes other than the 10-repeat allele and cognitive endophenotypes in ADHD. The aim of this study was to examine the relationship between the DAT1 genotypes and the candidate endophenotypes, inattention and impulsivity symptoms, as measured by the continuous performance test (CPT), in a Korean sample of 85 children diagnosed with DSM-IV ADHD. Compared to the normal control group, the frequencies of the 9/10 genotype were significantly higher in the ADHD probands (chi(2)=13.45, p=0.02, OR=4.12, 95% CI: 2.21-12.34) and parents of probands (chi(2)=11.60, p=0.03). The 9-repeat allele frequencies were significantly higher in the ADHD probands (chi(2)=11.55, p=0.03, OR=4.43, 95% CI: 1.55-11.78) and parents of probands (chi(2)=12.70,p=0.03) than the normal control group. Compared to the ADHD probands without the 9-repeat allele (n=74), the mean T-score, with regard to the commission errors of the CPT, was significantly higher (p<0.05) in the ADHD probands with the 9-repeat allele (n=11). Compared to the ADHD probands with other DAT1 genotypes, the mean T score, with respect to the commission errors of the CPT, was significantly higher in the ADHD probands with the 9/10 genotype (p<0.05). The results of this study suggest the possibility of an association between the DAT1 9-repeat allele and the impulsivity phenotype of ADHD. PMID- 16368112 TI - Some observations on the article "Dynamic response of cancer under the influence of immunological activity and therapy". PMID- 16368113 TI - A mathematical model of ion movements in grey matter during a stroke. AB - The development of cytotoxic oedema during a stroke consists in cell swelling and shrinking of the extracellular space. This phenomenon is triggered by ion movements through voltage-gated channels, exchangers and pumps. During ischaemia, sodium, calcium and chloride enter the neurons whereas potassium and glutamate are expelled out of the cells. A mathematical model is proposed to represent the long-term dynamics of membrane potentials, cell volumes and ionic concentrations in intracellular and extracellular spaces during a stroke and to study the influence of each ionic current on cell swelling. The model relies on electrophysiological mechanisms and takes into account the behaviour of two types of cells: neurons and also astrocytes known to play a key role in the excitotoxic process in grey matter. The results obtained when a severe or a moderate ischaemia is simulated are consistent with those observed in the in vitro and in vivo experiments. As this model appears to be robust, it is used to perform illustrative simulations aimed at studying the effect of some channel blockers on cell swelling. This approach may help to explore new therapeutic strategies in order to reduce stroke damage. PMID- 16368114 TI - Cyclic strain modulates tubulogenesis of endothelial cells in a 3D tissue culture model. AB - Angiogenesis is the formation of new blood vessels from preexisting capillaries or venules. It occurs in a mechanically dynamic environment due to blood flow, but the role of hemodynamic forces in angiogenesis remains poorly understood. We have developed a unique in vitro system for the investigation of angiogenesis under cyclic strain. In this system, tubulogenesis of vascular endothelial cells in 3D collagen gels occurs under well-defined cyclic strain, which mimics blood pressure-induced stretch. Using this system, we demonstrate that cyclic strain results in alignment of endothelial-cord-like structures perpendicular to the principal axis of stretch. Such preferential orientation was the most evident in deep and long cord-like structures. This in vitro system, along with the novel findings of strain-modulated endothelial tube morphology, enables the formation of an experimental basis for understanding the role of cyclic strain in the regulation of angiogenesis. PMID- 16368115 TI - Regional and cellular distribution of CYP2E1 in monkey brain and its induction by chronic nicotine. AB - CYP2E1 is expressed in liver and extrahepatic tissues, including brain. It metabolizes ethanol and other drugs and toxins, such as acetaminophen, chlorzoxazone and tobacco-derived nitrosamines. Hepatic CYP2E1 is inducible by nicotine, and cigarette smoke accelerates chlorzoxazone metabolism. Smokers have higher levels of brain CYP2E1 expression than non-smokers, but the specific regions and cell types which have the higher expression differ from nicotine induced rat brain. We therefore investigated the expression and distribution of brain CYP2E1 in a non-human primate, the African green monkey, and determined the effect of nicotine treatment. CYP2E1 levels varied among saline-treated monkey brain regions (P < 0.01) and expression was cell-type specific. Chronic nicotine treatment induced CYP2E1 expression in the frontal cortex (1.5-fold, P < 0.05) and cerebellum (1.5-fold, P < 0.01), specifically in cortical pyramidal neurons and cerebellar Purkinje cells but no change was seen in temporal cortex (P = 0.20), hippocampus (P = 0.29), putamen (P = 0.26) and thalamus (P = 0.08). CYP2E1 expression pattern in monkey brain following chronic nicotine treatment is similar to that in smokers, suggesting that nicotine may be the primary component in cigarette smoke that induces CYP2E1. Increased CYP2E1 in brain may contribute to oxidative stress and alter localized metabolism, and resulting pharmacology, of centrally acting drugs metabolized by CYP2E1. PMID- 16368116 TI - A behavioural and ERP investigation of 3-month-olds' face preferences. AB - Recent behavioural work suggests that newborns' face preferences are derived from a general, non-specific attentional bias toward patterns with more features in the upper versus lower half. In the current study, we predicted that selectivity for the specific geometry of the face may emerge during the first 3 months of life as a product of perceptual narrowing, leading to the construction of the first broadly defined face category segregating faces from other visual objects which may share with faces one or more visual properties. This was investigated behaviourally, using a standard preferential looking paradigm, and electrophysiologically, using high-density ERPs. Behavioural results indicated that, at 3 months, the top-heavy property is no longer a crucial factor in determining face preferences. ERP results showed evidence of differentiation between the two stimuli only for the N700. No differentiation was found for earlier components that are thought to reflect the adult-like structural encoding stage of face processing in infants (N290 and P400). Together, ERP and behavioural results suggest that, by 3 months, the perceptual narrowing process has led to a behavioural response specific to the geometry of the human face, but that this response is not purely perceptual in nature. Rather, it seems related to the acquired salience of this stimulus category, which may reflect the high degree of familiarity and/or the social value faces have gained over the infants' first 3 months of life. PMID- 16368117 TI - Action control in visual neglect. AB - Patients with unilateral neglect show a variety of impairments when reaching towards objects in contralesional space. The basis of these deficits could be perceptual, motor or at one of the intermediate stages linking these processes. Here, we review studies of visually guided reaching in neglect and integrate these results with findings from normal human and monkey action control. We consider evidence which shows that neglect patients can be slow to initiate or execute reaches particularly to a contralesional target. We discuss the directional and spatial deficits that may interact to contribute to such reaching abnormalities and highlight the importance of effective target selection and on line guidance, exploring the idea that deficits in these mechanisms underlie increased susceptibility to ipsilesional visual distraction in neglect. We also examine the relationship between optic ataxia and neglect by considering two illustrative cases, one with pure optic ataxia and the other with optic ataxia plus neglect, which reveal differences in the anatomical substrates of the two syndromes. We conclude that many patients with neglect make abnormal visually guided reaches, but the pattern of reaching deficits is highly variable, most likely reflecting heterogeneity of lesion location across subjects. Rather than being specific to the neglect syndrome, abnormalities of reaching in these patients may correspond to the extent of damage to the visuomotor control system which involves critical regions in both the parietal and frontal cortex, the white matter tracts connecting them and subcortical regions. Thus, the action control deficits in neglect may be conceptualised as a range of impairments affecting multiple stages in the visuomotor control process. PMID- 16368118 TI - Bitter taste markers explain variability in vegetable sweetness, bitterness, and intake. AB - Intake of vegetables falls short of recommendations to lower risk of chronic diseases. Most research addresses bitterness as a sensory deterrent to consuming vegetables. We examined bitter and sweet sensations from vegetables as mediators of vegetable preference and intake as well as how these tastes vary with markers of genetic variation in taste (3.2 mM 6-n-propylthiouracil bitterness) and taste pathology (1.0 mM quinine bitterness, chorda tympani nerve relative to whole mouth). Seventy-one females and 39 males (18-60 years) reported prototypical tastes from and preference for Brussels sprouts, kale and asparagus as well as servings of vegetables consumed, excluding salad and potatoes. Intensity and hedonic ratings were made with the general Labeled Magnitude Scale. Data were analyzed with multiple linear regression and structural equation modeling. Vegetable sweetness and bitterness were independent predictors of more or less preference for sampled vegetables and vegetable intake, respectively. Those who taste PROP as most bitter also tasted the vegetables as most bitter and least sweet. The spatial pattern of quinine bitterness, suggestive of insult to chorda tympani taste fibers, was associated with less bitterness and sweetness from vegetables. Via structural equation modeling, PROP best explained variability in vegetable preference and intake via vegetable bitterness whereas the quinine marker explained variability in vegetable preference and intake via vegetable bitterness and sweetness. In summary, bitterness and sweetness of sampled vegetables varied by taste genetic and taste function markers, which explained differences in preference for vegetables tasted in the laboratory as well as overall vegetable intake outside the laboratory. PMID- 16368119 TI - Cytotoxic and aromatic constituents from Salvia miltiorrhiza. AB - As part of an ongoing study of traditional Chinese medicinal plants, the root tissue of Salvia miltiorrhiza was further investigated for its chemical constituents. Five naturally occurring products along with 13 known constituents were isolated from an ethyl acetate-soluble portion of its ethanol extract. Their structures were elucidated by means of spectroscopic methods. Some selected compounds were also evaluated for biological activity. PMID- 16368120 TI - Anti-plasmodial and antioxidant activities of constituents of the seed shells of Symphonia globulifera Linn f. AB - A xanthone derivative, named gaboxanthone (1), has been isolated from the seed shells of Symphonia globulifera, together with known compounds, symphonin (2), globuliferin (3), guttiferone A (4), sistosterol, oleanolic acid and methyl citrate. The structure of the compound was assigned as 5,10-dihydroxy-8,9 dimethoxy-2,2-dimethyl-12-(3-methylbut-2-enyl) pyrano [3,2-b]xanthen-6(2H)-one, by means of spectroscopic analysis. The anti-plasmodial and antioxidant activities of the phenolic compounds were evaluated, respectively, in culture against W2 strain of Plasmodium falciparum and using the free radical scavenging activity of the DPPH radical, respectively. Compounds 1-4 were found to be active against the Plasmodium parasites (IC(50) of 3.53, 1.29, 3.86 and 3.17 microM, respectively). Guttiferone A (4) showed a potent free radical scavenging activity compared to the well-known antioxidant caffeic acid. PMID- 16368121 TI - One-pot, high yield synthesis of alpha-ketols from Delta5-steroids. AB - Alpha-hydroxy ketones (alpha-ketols) are present in many compounds with biological activity. Several methods are available for the preparation of alpha ketols but only a few of them describe the synthesis of steroid alpha-ketols from olefins. In this work, a new system consisting of KMnO(4)/Fe(ClO(4))(3).nH(2)O was used in order to achieve the direct conversion of Delta(5)-steroids to their corresponding alpha-ketols, in high yields. Consideration of the probable reaction mechanism is provided. 2D homo- and heteronuclear correlation NMR spectroscopic techniques were used to assign (1)H and (13)C resonances of some synthesized compounds. This method has potential for the preparation of alpha hydroxy ketones of biological interest. PMID- 16368122 TI - Transplacental arsenic plus postnatal 12-O-teradecanoyl phorbol-13-acetate exposures associated with hepatocarcinogenesis induce similar aberrant gene expression patterns in male and female mouse liver. AB - Our prior work shows that in utero arsenic exposure alone is a complete transplacental carcinogen, producing hepatocellular carcinoma in adult male offspring but not in females. In a follow-up study to potentially promote arsenic initiated tumors, mice were exposed to arsenic (85 ppm) from gestation day 8 to 18 and then exposed to 12-O-teradecanoyl phorbol-13-acetate (TPA), a well-known tumor promoter after weaning. The dermal application of TPA (2 mug/0.1 ml acetone, twice/week for 21 weeks) after transplacental arsenic did not further increase arsenic-induced liver tumor formation in adult males but significantly increased liver tumor formation in adult females. Thus, for comparison, liver tumors and normal liver samples taken from adult male and female mice at necropsy were analyzed for aberrant gene/protein expression by microarray, real-time RT PCR and Western blot analysis. Arsenic/TPA treatment resulted in increased expression of alpha-fetoprotein, k-ras, c-myc, estrogen receptor-alpha, cyclin D1, cdk2na, plasminogen activator inhibitor-1, cytokeratin-8, cytokeratin-18, glutathione S-transferases and insulin-like growth factor binding proteins in liver and liver tumors from both male and female mice. Arsenic/TPA also decreased the expression of BRCA1, betaine-homocysteine methyltransferase, CYP7B1, CYP2F2 and insulin-like growth factor-1 in normal and cancerous livers. Alterations in these gene products were associated with arsenic/TPA-induced liver tumors, regardless of sex. Thus, transplacental arsenic plus postnatal TPA exposure induced similar aberrant gene expression patterns in male and female mouse liver, which are persistent and potentially important to the mechanism of arsenic initiation of hepatocarcinogenesis. PMID- 16368123 TI - Comparison of effects of estradiol with those of octylmethoxycinnamate and 4 methylbenzylidene camphor on fat tissue, lipids and pituitary hormones. AB - Octylmethoxycinnamate (OMC) and 4-methylbenzylidene camphor (4MBC) are commercially used absorbers of ultraviolet (UV) light. In rats, they were shown to exert endocrine disrupting including uterotrophic, i.e. estrogenic effects. Estrogens have also metabolic effects, therefore the impact of oral application of the two UV absorbers at 2 doses for 3 months on lipids and hormones were compared with those of estradiol-17beta (E2). E2, OMC and 4MBC reduced weight gain, the size of fat depots and serum leptin, a lipocyte-derived hormone, when compared to the ovariectomized control animals. Serum triglycerides were also reduced by the UV screens but not by E2. On the other hand, E2 and OMC reduced serum cholesterol, low density lipoproteins and high density lipoproteins; this effect was not shared by 4MBC. While E2 inhibited, OMC and 4MBC stimulated serum LH levels. In the uterus, both UV filters had mild stimulatory effects. 4MBC inhibited serum T4 resulting in increased serum TSH levels. It is concluded that OMC and 4MBC have effects on several metabolic parameters such as fat and lipid homeostasis as well as on thyroid hormone production. Many of these effects are not shared by E2. Hence, other than estrogen-receptive mechanisms may be responsible for these effects. PMID- 16368124 TI - Human herpesvirus 6 (HHV-6) U94/REP protein inhibits betaherpesvirus replication. AB - Human herpesvirus 6 (HHV-6) is the only human herpesvirus encoding U94/rep, homologue to the parvovirus non-structural gene rep68/78. Results to date suggest that HHV-6 U94/rep might regulate viral gene expression and have a role in viral latency. To determine the effect of U94/REP upon viral replication, the protein was produced. The purified U94/REP retained the characteristic immunological features. It was internalized and localized in the nucleus of human cells, showing marked inhibitory activity on the replication of HHV-6 (both variants A and B). The effect of U94/REP was dose-dependent and sensitive to treatment with single-stranded but not double-stranded DNA. U94/REP inhibited the replication of other betaherpesviruses, HHV-7 and human cytomegalovirus, but had no effect on herpes simplex virus. These results confirm the action of U94/rep latency gene in the regulation of HHV-6 replication with implications for co-reactivations and latency of human betaherpesviruses. PMID- 16368125 TI - gC1qR expression in chimpanzees with resolved and chronic infection: potential role of HCV core/gC1qR-mediated T cell suppression in the outcome of HCV infection. AB - Chimpanzee is a unique animal model for HCV infection, in which about 50% of infections resolve spontaneously. It has been reported that the magnitude of T cell responses to HCV core in recovered chimpanzees is greater than that in chronically infected ones. However, the mechanism(s) by which the chimpanzees with resolved infection overcome core-mediated immunosuppression remains unknown. In this study, we examined the effect of HCV core on T cell responsiveness in chimpanzees with resolved and chronic HCV infection. We found that core protein strongly inhibited T cell activation and proliferation in chimpanzees with chronic infection, while this inhibition was limited in chimpanzees with resolved infection. Notably, the level of gC1qR, as well as the binding of core protein, on the surface of T cells was lower in recovered chimpanzees when compared to chimpanzees with chronic HCV infection. Intriguingly, the observed differences in gC1qR expression levels and susceptibility to core-induced suppression amongst HCV-chronically infected and recovered chimpanzees were observed prior to HCV challenge, suggesting a possible genetic determination of the outcome of infection. These findings suggest that gC1qR expression on the surface of T cells is crucial for HCV core-mediated T cell suppression and viral clearance, and that represents a novel mechanism by which a virus usurps host machinery for persistence. PMID- 16368126 TI - Exogenous attention and endogenous attention influence initial dominance in binocular rivalry. AB - We investigated the influence of exogenous and endogenous attention on initial selection in binocular rivalry. Experiment 1 used superimposed +/-45 degrees gratings viewed dioptically for 3s, followed by a brief contrast increment in one of the gratings to direct exogenous attention to that grating. After a brief blank period, dichoptic stimuli were presented for various durations (100-700 ms). Exogenous attention strongly influenced which stimulus was initially dominant in binocular rivalry, replicating an earlier report (Mitchell, Stoner, & Reynolds. (2004). Object-based attention determines dominance in binocular rivalry. Nature, 429, 410-413). In Experiment 2, endogenous attention was manipulated by having participants track one of two oblique gratings both of which independently and continuously changed their orientations and spatial frequencies during a 5s period. The initially dominant grating was most often the one whose orientation matched the grating correctly tracked using endogenous attention. In Experiment 3, we measured the strength of both exogenous and endogenous attention by varying the contrast of one of two rival gratings when attention was previously directed to that grating. The contrast of the attended grating had to be reduced by an amount in the neighborhood of 0.3 log-units, to counteract attention's boost to initial dominance. Evidently both exogenous and endogenous attention can influence initial dominance of binocular rivalry, effectively boosting the stimulus strength of the attended rival stimulus. PMID- 16368127 TI - Levels of non-essential elements in muscle from harp seal (Phagophilus groenlandicus) and hooded seal (Cystophora cristata) caught in the Greenland Sea area. AB - The non-essential elements, arsenic, cadmium, mercury and lead, inevitably accumulate in marine top predators such as seals. The concentration of these elements and the essential element selenium, due to its proposed protective properties against mercury toxicity in marine mammals, were measured in muscle, liver and kidney from reproductive active females of harp seal (Phagophilus groenlandicus) and hooded seal (Cystophora cristata) caught in the drift ice between Iceland and East Greenland. Arsenic levels were below 1 microg/g w.w. in all analysed samples, and were therefore low compared to other seafood products. The concentrations of arsenic found in the present study were comparable to the results reported in a similar study from 1985. Mean concentrations of total mercury in muscle from the present study were higher than levels in other seafood products. The levels of total mercury from the present study showed a tendency of lower levels in all tissue samples compared to the study from 1985. Methyl mercury displayed a trend of a lower ratio of methyl mercury to total mercury as the concentration of total mercury increased, indicating a demethylation of methyl mercury at high total mercury concentrations (e.g. mercury in liver of hooded seal). The concentration ratio of methyl mercury to total mercury in muscle samples was more than 75%, with total mercury concentration less than 0.5 microg/g w.w., whereas the ratio for liver was as low as 0.2% with a total mercury concentration of 128 microg/g w.w. The molar concentration ratios of selenium to mercury showed that selenium was present in a molar surplus to mercury in all tissues with low mercury concentration. However, there seemed to be a general mobilisation of selenium in liver and kidney tissues of harp seal and hooded seal, whereas an extraordinary mobilisation seemed to take place at hepatic mercury concentrations exceeding 50 microg/g w.w. The mean concentrations of lead in muscles in the present study were higher than in fish and other seafood products from the Barents Sea. The lead concentrations from the present study were lower than levels reported in the 1985 study. However, the levels of the non-essential elements analysed in muscle from the two seal species in the present study should not prevent the use of seal meat in human nutrition. PMID- 16368128 TI - Harm or benefit of hormonal treatment in metastatic low-grade endometrial stromal sarcoma: single center experience with 10 cases and review of the literature. AB - Endometrial stromal sarcoma (ESS) is a rare disease with probably less than 700 new cases in the US or EU per year. ESS usually expresses steroidal receptors and is regarded to be hormone-sensitive. A higher risk in women receiving estrogen replacement therapy (ERT) or tamoxifen has been suspected, and remissions following treatment with progestins have been reported in case studies. Aromatase inhibitors represent an interesting new treatment option. Due to the rarity of the tumor, only few case series and no prospective studies are published. We therefore conducted a retrospective study to evaluate the influence of hormonal treatment to ESS. METHODS: Our institutional sarcoma data bank was screened for cases of ESS since 1999. All corresponding files and radiographs were reviewed retrospectively. RESULTS: Ten patients with low-grade ESS were identified. Diagnosis was established before or by hysterectomy in 6 patients, by local recurrence after previous hysterectomy for nonmalignant disease in 3 patients or by pulmonary metastases with no primary tumor found so far in 1 patient. 5/10 patients were on ERT and 3/10 on tamoxifen at the time of diagnosis of metastatic disease. Treatment strategies consisted of stopping ERT and tamoxifen, respectively, or initiation of the progestin MPA or letrozole. Three patients achieved stable disease after stopping ERT. 2/3 patients responded to MPA as first-line treatment (1 CR; 50+ months, 1 PR; 9 months). 4/5 patients responded to letrozole as first-line therapy (3 PR;3+, 9+ and 10+ months) or second-line treatment after MPA (1 PR; 37+ months). 9/10 patients are alive 33 to 255 months after hysterectomy. Survival since diagnosis of metastatic disease is 4 to 164 months. CONCLUSIONS: Patients with a previous history of low-grade ESS should not be treated with estrogens or tamoxifen. If nevertheless present, withdrawal of ERT or tamoxifen is strongly advised, resulting in disease stabilization in some cases. MPA and letrozole, in particular, are highly effective and lead to sustained disease control in most cases. PMID- 16368129 TI - Lack of behavioral tolerance by repeated treatment with taltirelin hydrate, a thyrotropin-releasing hormone analog, in rats. AB - In order to determine whether acute tolerance develops by taltirelin hydrate ((-) N-[(S)-hexahydro-1-methyl-2,6-dioxo-4-pyrimidinylcarbonyl]-l-histidyl-l prolinamide tetrahydrate; taltirelin), a thyrotropin-releasing hormone (TRH) analog, we examined the motor behavior, TRH receptors and dopamine D(2) receptors following 2 weeks treatment in rats. Taltirelin selectively bound to TRH receptors and increased the spontaneous motor activity by a single administration, suggesting that the motor effect of taltirelin is mediated by TRH receptors. Following repeated treatment with TRH, there was a significant reduction in the increment of spontaneous motor activity. In contrast, after repeated treatment with taltirelin at a dose that increased the motor activity to a similar extent to TRH by a single administration, there was no apparent change in its motor effect. In accord with the motor activity, we found a significant reduction in the [(3)H]methyl-TRH binding to TRH receptors in the brain following repeated treatment with TRH but not taltirelin. However, the [(3)H]spiperone binding to dopamine D(2) receptors in the corpus striatum did not change by repeated taltirelin and TRH treatments. Thus, the down-regulation of TRH receptors would be a main cause of the behavioral tolerance. These results suggest that taltirelin hardly develops the behavioral tolerance due to the lack of down-regulation of TRH receptors. PMID- 16368130 TI - Influence of cavity dimensions and their derivatives (volume and 'C' factor) on shrinkage stress development and microleakage of composite restorations. AB - OBJECTIVES: To determine, in vitro, the influence of cavity dimensions on shrinkage stress development and microleakage of composite restorations. METHODS: Cylindrical cavities with 2, 3, 4 or 6 mm diameter and 1 or 2 mm depth were prepared in bovine incisors (with enamel margins) and reproduced in photoelastic resin. Cavities were restored in bulk (Single Bond+Filtek Z250). Stress fringes were analyzed 10 min after photoactivation. Restored teeth were stored for 24 h, coated with nail polish and immersed in 50% AgNO3 for 2 h, followed by 6 h in developing solution. Specimens were sectioned twice and microleakage was measured under 20x magnification. Photoelastic data were analyzed descriptively only, due to lack of variability in some groups. Microleakage was analyzed through (generalized linear) regression models. Pearson correlation coefficients were computed to evaluate the relationship between microleakage and 'C' factor or volume. RESULTS: Fringe order was higher at the internal angles than at the margins of the restorations. There was a trend for higher fringe orders with increasing diameter and depths. Regression analysis revealed a significant influence of restoration depth for diameters above 2mm. The effect of diameter on microleakage is significantly higher (p < 0.0001) for 2-mm deep restorations, compared to those 1-mm deep. Pearson correlation coefficients suggest some relationship between microleakage and volume (r = 0.724, p < 0.0001), but not between microleakage and 'C' factor (r = 0.048, p = 0.6120). CONCLUSIONS: Shrinkage stress and microleakage were higher in restorations with larger diameters and depths. Microleakage seemed to be related to a restoration's volume, but not to its 'C' factor. PMID- 16368131 TI - Interphase structure-property relationships in thermoset dimethacrylate nanocomposites. AB - OBJECTIVES: The aim of this research was to better understand the relationships between interphase composition and the resultant mechanical properties of thermoset methacrylate nanocomposites, ultimately for the purpose of improving the properties of dental restorative materials through manipulation of the interphase. METHODS: Silica nanoparticles were silanized with three different silanes and blends of those silanes to generate six different interphase compositions. The silanes varied in their relative polarity, flexibility, and reactivity towards photo-polymerization. Composites containing 60% by mass of the silanized fillers were prepared and analyzed for uncured paste handling characteristics, vinyl conversion, biaxial flexure strength (BFS), modulus, and Knoop hardness. RESULTS: Dual silanization of the fillers improved the handling characteristics of the uncured composite pastes compared to those containing fillers silanized with 3-methacryloxypropyltrimethoxysilane. To obtain high BFS, functional groups reactive in free radical polymerization were needed in the composite interphase, but a high concentration of those groups was not necessary. Moduli were highest for composites with interphases that contained styrylethyltrimethoxysilane, a reactive aromatic silane. The hardness values of the composites with reactive interphases were all comparable. Methacrylate conversion was only modestly influenced by silane interphase composition. SIGNIFICANCE: This study suggests that dual silanization is a practical method for improving the handling characteristics of uncured dental restorative nanocomposites while maintaining or improving the mechanical properties of the cured composites. PMID- 16368132 TI - The effect of consistency, specimen geometry and adhesion on the axial polymerization shrinkage measurement of light cured composites. AB - OBJECTIVES: This study investigated the effect of the consistency, specimen geometry and adhesion on the measurements of axial polymerization shrinkage of light cured composite resins using an axial shrinkage-measuring device. METHODS: Four commercially available composites were examined: an anterior posterior hybrid composite Z100, a posterior packable composite P60 and two flowable composites, Filtek flow and Tetric flow. The axial polymerization shrinkage of the composites was determined using a 'bonded disc method' and 'non-bonded' free shrinkage method at varying C-factors by altering the specimen geometry. These measured axial shrinkages were compared with the free volumetric shrinkages. The consistency of the composites was also compared using a squeeze test. RESULTS: Using the non-bonded method, the axial shrinkage was approximately one third of the true volumetric shrinkage as a result of isotropic contraction. However, in the bonded disc method, the axial shrinkage increased up to the volumetric shrinkage by anisotropic contraction as the bonded surface increased. The axial shrinkage increased with the increasing C-factor. It approached the true volumetric shrinkage and reached a plateau at near a C-factor of 5-6. However, in flowable composites, a lower level of axial shrinkage was measured by the compensational radial flow. SIGNIFICANCE: When estimating the volumetric shrinkage from the axial shrinkage measured using the bonded disc method, the C factor of the specimen should be higher than 5 and the consistency of the composite is also an important factor that needs to be considered. PMID- 16368133 TI - Catalytic efficiency of a thrombomodulin-functionalized membrane-mimetic film in a flow model. AB - The protein C anticoagulant pathway generates an "on demand" physiologic anticoagulant response, which is initiated when thrombin binds to thrombomodulin (TM), a transmembrane protein constitutively expressed by endothelial cells. A stable, protein C activating membrane-mimetic film was produced on a polyelectrolyte multilayer (PEM) by in situ photopolymerization of a phospholipid assembly containing TM. The monoacrylated phospholipid monomer was initially synthesized and prepared as unilamellar vesicles with varying molar concentrations of TM. Membrane-mimetic films were constructed on planar substrates with defined surface concentrations of catalytically active TM. 125I labeled radiolabeling demonstrated little change in TM surface concentration over periods of up to 4 weeks. We utilized a parallel plate flow system to investigate the effects of simulated arterial (500 s(-1)) and venous (50 s(-1)) shear rates and TM surface concentration (0-1400 fmol cm(-2)) on the rate and extent of activation of protein C. The rate of production of activated protein C increased with shear rate and TM surface content. However, in agreement with an analysis of reaction kinetics and mass transfer, experimental results demonstrate that reaction rates become saturated at TM surface densities greater than or equal to 800 fmol cm(-2). We believe that the design of membrane-mimetic films that have the capacity to activate the protein C pathway will provide a useful strategy for generating "actively" antithrombogenic surfaces. PMID- 16368134 TI - Stem cell-coated titanium implants for the partial joint resurfacing of the knee. AB - The goal of the present study was to evaluate the partial surface replacement of the knee with stem cell-coated titanium implants and to provide a basis for a successful treatment of large osteochondral defects. Mesenchymal stem cells (MSCs) were isolated from bone marrow aspirates of adult sheep. Round titanium implants with a diameter of 2 x 7.3 mm were seeded with autologous MSC and inserted into an osteochondral defect in the medial femoral condyle. As controls, defects received either an uncoated implant or were left untreated. Nine animals with 18 defects were sacrificed after 6 months. Histological evaluation was performed by intravital polychrome fluorescent labelling, intravital perfusion with Indian ink, microradiographs and differential staining with toluidine blue. The quality of regenerated cartilage was assessed by in situ hybridization of collagen type II and immunohistochemistry of collagen types I and II. In 50% of the cases, defects treated with MSC-coated implants showed a complete regeneration of the subchondral bone layer. In these cases collagen type II and only traces of collagen type I were detected. A high level of collagen type II mRNA expression compared to articular cartilage indicates regenerating hyaline like cartilage. A total of 50% of MSC-coated and uncoated implants failed to osseointegrate and formation of fibrocartilage was observed. Untreated defects as well as defects treated with uncoated implants demonstrated incomplete healing of subchondral bone and formation of fibrous cartilage. A modified histological score according to Wakitani significantly demonstrated better results for cell coated implants (8.8+/-6.4) than for uncoated implants (5.5+/-3.9) and for untreated defects (2.8+/-2.5). Our results demonstrate that, in a significant number of cases, a partial joint resurfacing of the knee with stem cell-coated titanium implants occur. A slow bone and cartilage regeneration and an incomplete healing in half of the MSC-coated implants are limitations of the presented method. To improve our approach and optimize the experimental parameters, further investigations are needed prior to clinical application. PMID- 16368135 TI - Fabrication and characterization of heparin functionalized membrane-mimetic assemblies. AB - A membrane-mimetic assembly incorporating surface bound heparin was fabricated as a system to improve the hemocompatibility of blood-contacting devices. As a model system, heparin was chemically modified by end-point conjugation to biotin and immobilized onto membrane-mimetic thin films via biotin-streptavidin interactions. Heparin surface density, determined by radiochemical titration, confirmed that surface density was directly related to the molar concentration of biotinylated lipid within the assembled membrane-mimetic film. The capacity of surface bound heparin to promote ATIII-mediated thrombin inactivation was investigated in a parallel plate flow chamber under simulated venous and arterial wall shear rates of 50 and 500 s(-1), respectively. Significantly, we observed that the rate of thrombin inactivation approached a maximum at a heparin surface concentration greater than 4.4 pmol/cm(2) (61 ng/cm(2)). In the process, mass transport limited regimes were identified for heparin potentiated thrombin inactivation under both simulated venous and arterial conditions. PMID- 16368136 TI - Voltage-gated calcium channels mediate hypoxic vasoconstriction in the human placenta. AB - Unlike all vascular beds with the exception of the pulmonary circulation, fetoplacental vessels respond to acute hypoxia with vasoconstriction. While this hypoxic fetoplacental vasoconstriction (HFPV) is considered essential in the pathogenesis of intrauterine growth retardation, its mechanism is largely unknown. Hypoxia inhibits potassium channels and thus causes depolarization in fetoplacental vascular smooth muscle. We propose that this hypoxia-induced depolarization leads to vasoconstriction by activating voltage-dependent calcium (Ca) channels and Ca influx. We compared HFPV between isolated perfused human cotyledons treated with an inhibitor of L-type channels, nifedipine, and preparations receiving only vehicle. While the solvent (diluted DMSO) had no inhibitory effect on HFPV, the hypoxic responses were completely abolished even by a relatively low dose of nifedipine (1 nM). We conclude that activation of L type Ca channels is an essential part of HFPV. PMID- 16368137 TI - Non-sense mutations in the dihydropyridine receptor beta1 gene, CACNB1, paralyze zebrafish relaxed mutants. AB - Contractions by skeletal muscle require proper excitation-contraction (EC) coupling, whereby depolarization of the muscle membrane leads to an increase in cytosolic Ca(2+) and contraction. Changes in membrane voltage are detected by dihydropyridine receptors (DHPR) that directly interact with and activate ryanodine receptors to release Ca(2+) from the sarcoplasmic reticulum into the cytosol. A genetic screen for motility mutations isolated a new allele of the immotile zebrafish mutant relaxed. Muscles in relaxed embryos do not contract in response to potassium chloride (KCl) thus appear unresponsive to membrane depolarization, but do contract when stimulated by caffeine, an agonist of ryanodine receptors. This suggests that relaxed mutant muscles are defective in EC coupling. Indeed, immunohistochemical analysis demonstrated that mutants lack DHPRs in skeletal muscles. The mutant phenotype results from non-sense mutations in the zebrafish CACNB1 gene that encodes the DHPR beta1 subunit. The zebrafish CACNB1 gene is expressed in skeletal muscles, PNS and CNS. Electrophysiological recordings showed no obvious abnormalities in the motor output of relaxed mutants, presumably due to redundancy provided by other beta subunits. The structural and functional homology of CACNB1 in zebrafish and mammals, suggests that zebrafish can be useful for studying EC coupling and potential neuronal function of CACNB1. PMID- 16368138 TI - Down-regulation of antibacterial peptide synthesis in an insect model induced by the body-surface of an entomoparasite (Steinernema feltiae). AB - This study focuses on the interaction between an entomopathogenic nematode and an insect model to further investigate the role of parasite body-surface and its compounds (particularly epicuticular lipids) in the immunosuppression of host defences. Our goal was to ascertain the potential interference of the parasite epicuticular lipids with the antimicrobial response mounted by Gram-negative bacteria-challenged hosts. Since the parasite model used in this study (Steinernema feltiae) releases symbiontic bacteria in the host hemocoel during the late phase of infection, the inhibition of the antimicrobial response could be needed to avoid the activity of host factors potentially harmful for the microorganisms. After bacterial challenge, when insect hosts were infected with purified parasite cuticles, we always observed lack of bacterial clearance concurrently with the absence of hemolymph low molecular weight components. The observed effects seem to be related to the interaction of parasite cuticular lipids (PCLs) with specific components of the host hemolymph; these host interacting proteins (HIP17, HIP26 and HIP35) were removed by the parasite, and their absence (or reduction) apparently prevented antimicrobial peptide synthesis. The inhibitory properties were lost when cuticles were pre-treated with compounds (such as lipase or methanol-chloroform) affecting their lipidic moiety. Moreover, the key role of epicuticular lipids was also confirmed by the inhibitory properties of methanol-chloroform extracted lipids, which were comparable to those of parasite whole cuticles. Finally, the involvement of HIPs was assessed by their partial purification followed by injection into the host. When HIPs were co-injected with bacteria into cuticle-inhibited larvae, the antimicrobial activity was completely restored. PMID- 16368139 TI - Detection of QTL for innate: non-specific antibody levels binding LPS and LTA in two independent populations of laying hens. AB - In the current study results are presented of an experiment dealing with the Natural antibodies which are measured by level of homotopes LPS and LTA. Two independent populations were examined (F2 population descendant from a cross between chickens divergently selected for either High or Low specific Ab responses to SRBC (HL) and F2 cross descendant from lines expressed different behavior concerning feather pecking (FP)). In total 12 QTL were detected to non specific antibody titers directed to LTA and LPS and at two ages after applying two statistical models in an F2 HL population. Similarly in an FP cross overall seven QTL were detected. Based on obtained results it might be concluded that different QTL regions are associated with immune responses to homotopes LPS and LTA in poultry. PMID- 16368140 TI - TNF induces the growth of thymocytes in rainbow trout. AB - In order to investigate the effects of TNFalpha upon the growth of fish thymocytes, rainbow trout thymocytes were cultured in the conditioned medium (CM): the supernatants of the macrophage cultures stimulated with chitin derivative and LPS. Synthesis of TNFalpha by macrophages and subsequent secretion into CM were ascertained by RT-PCR and western blotting. While most of the thymocytes cultured in normal medium died within 7 days, the thymocytes cultured in CM exhibited markedly better growth as monitored by alamarBlue assay and BrdU assay. The proliferating cells appeared to be small lymphocytes. Since such activity in CM was significantly inhibited by an anti-trout TNF antibody, it was clearly evident that TNFalpha in the CM induced the proliferation of the thymocytes. Production of TNFalpha in the thymus of healthy fish was also demonstrated by RT-PCR. Collectively, this data suggest that TNFalpha is involved in T cell development in the trout thymus. PMID- 16368141 TI - Phytotoxicity and accumulation of anthracene applied to the foliage and sandy substrate in lettuce and radish plants. AB - The effects of anthracene (ANT) on the growth of two species of vegetable plants (Lactuca sativa L. and Raphanus sativus L.), which play an important role in the human diet, were studied. ANT was applied to the leaves of these plants by foliar deposition, in aerosol form, and to the sandy substrate in which the plants were grown in a greenhouse. It was found that ANT affected plant biomass, especially root biomass, in the case of both foliar and soil application. Under conditions of induced chemical stress, the dry matter of aboveground parts and roots was lower than that in control plants. The rate of photosynthesis decreased by about 20% in both plant species following foliar ANT application. A lower rate of transpiration was also observed in lettuce plants. After the foliar application of ANT, small quantities of the compound were found in the leaves only (0.06 0.18% of the total dose). ANT translocation to other parts of the plants was not observed. This compound underwent rapid chemical changes on the leaf surface under greenhouse conditions. After the application of ANT to a sandy substrate, this compound was detected in the roots and aboveground parts of plants, which indicates that it was transported throughout the plant. In a sandy substrate, the process of ANT decomposition was much slower-60-70% of the administered dose was measured in the soil after the completion of the experiment. PMID- 16368142 TI - Heavy metal concentrations in ground beetles, leaf litter, and soil of a forest ecosystem. AB - The objective of this study was to quantify the relationships between heavy metal concentrations in soil, leaf litter, and ground beetles at four sampling sites of a forest ecosystem in Medvednica Nature Park, Croatia. Ground beetles were sampled by pitfall trapping. Specimens were dry-ashed and soil and beetle samples digested with nitric acid. Lead, cadmium, copper, zinc, manganese, and iron were analyzed using atomic absorption spectrometry. Statistically significant differences between plots were found for lead, cadmium, and iron in ground beetles. Correlations between ground beetles and soil or leaf litter were positive for lead and cadmium concentrations and negative for iron concentration. Differences in species metal concentrations were recorded. Higher concentrations of all studied metals were found in female beetles. However, a significant difference between sexes was found only for manganese. Significant differences in species metal concentrations were found for species that differ in feeding strategies and age based on breeding season and emergence of young adults. PMID- 16368143 TI - Determination of short-term copper toxicity in a multispecies microalgal population using flow cytometry. AB - This study was conducted to determine the role of algal-algal interactions in a multispecies microalgal population on their sensitivities to copper based on an enzyme inhibition assay using flow cytometric measures. Autofluorescence (chlorophyll a and phycocyanin) was used to identify species and count algal signals. The effect of multispecies population on copper toxicity of Microcystis aeruginousa was detected (1) at the same initial cell density, (2) at the same surface area, and (3) in the presence and absence of Chlorella pyrenoidosa and Scenedesmus obliquus. As copper concentrations increased, esterase activity of M. aeruginosa changed in a concentration-dependent manner. The 24 h EC(50) value of M. aeruginosa in the multispecies population was significantly (P < 0.05) higher than those in the single-species population. Compared with S. obliquus, the effect of C. pyrenoidosa on M. aeruginosa was more marked (the 24 h EC(50) value of copper on fluorescin diacetate fluorescence of M. aeruginosa was 11 microg/L). At 48 h copper exposure (6 microg/L) analysis of intracellular reactive oxygen species levels also showed similar algal-algal interactions in multispecies microalgal populations. The pigment assay suggested that these algal-algal interactions occurred only at low concentrations (< 13 microg/L, 24 and 48 h copper exposure). This study demonstrates the importance of using multispecies populations to estimate metal toxicity in natural waters. PMID- 16368144 TI - A possible new bridge between innate and adaptive immunity: Are the anti mitochondrial citrate synthase autoantibodies components of the natural antibody network? AB - Natural antibody (nAb) producing B-1 B cells are considered an intermediate stage of evolution between innate and adaptive immunity. nAbs are immunoglobulins that are produced without antigen priming. nAbs can recognize foreign targets and may serve in the first line of immune defense during an infection. Natural autoantibodies (nAAbs) present in the serum of both healthy humans and patients suffering from systemic autoimmune diseases recognize a set of evolutionarily conserved self-structures. Because of their endosymbiotic evolutionary origin, proteins compartmentalized into mitochondria represent an interesting transition from prokaryotic foreign (non-self) to essential (self) molecules. We investigated the possible overlap in recognized epitopes of innate and self reactive nAbs and surveyed changes in physiological autoreactivity under pathological autoimmune conditions. Epitope mapping analysis of a mitochondrial inner membrane enzyme, citrate synthase (CS) (EC 2.3.3.1) by synthetic overlapping peptides and phage display libraries using sera from healthy individuals and from patients having systemic autoimmune disease revealed CS recognizing nAAbs with IgM isotype. We analyzed cross reactive epitopes on human CS, bacterial CS, and various standard autoantigens. The anti-CS nAAbs by participating in the nAb network, could function in innate defense mechanisms and at the same time recognize a target antigen (nucleosome) in a systemic autoimmune disease. Thus, at the level of recognized epitopes there is a possible new link between the innate like component and the adaptive-autoimmune arm of the humoral immune system. PMID- 16368145 TI - Pregnancy-linked suppression of TcR signaling pathways by a circulating factor absent in recurrent spontaneous pregnancy loss (RPL). AB - Evidence suggests that maternal cell-mediated immunity is suppressed during pregnancy and that failure to suppress immune responses can result in partial or total rejection of the fetus. The molecular events associated with suppression of maternal T-cell activation mediated by circulating pregnancy-associated 14 kDa zeta inhibitor protein (ZIP) were defined in women with and without histories of recurrent pregnancy loss (RPL). Using cDNA microarray analysis, ZIP modulations of specific genes associated with T-cell activation signaling were defined. Alterations of defined components were confirmed at the protein level using chromatographically purified ZIP from normal pregnancies versus analogous material from women experiencing RPL. Based on microarray analyses, ZIP from normal pregnancies induced an increase (> or =2-fold) in the expression of 19 genes and a decrease (> or =2-fold) in 15 genes, when incubated with cultured T cells. In contrast, when T-cells were incubated with analogous material from RPL or non-pregnant controls, no significant differences were observed in the expression of these genes. At the protein level, ZIP from normal pregnancies induced decreases in CD3-zeta (2.36-fold), JAK3 (2.41-fold), STAT5 (1.85-fold), and NF-kappaB (4.24-fold) and a 2.05-fold increase in SOCS2 (all at p<0.001 compared to RPL and non-pregnant controls). The suppressive effects of Zip can lead to the failure of T-cell production of Th1 cytokines, such as IL-2. The 14 kDa circulating ZIP from normal pregnancies suppressed components within the JAK/STAT pathway and induced suppressors of cytokine stimulation, SOCS2. PMID- 16368146 TI - Anecortave acetate (15 milligrams) versus photodynamic therapy for treatment of subfoveal neovascularization in age-related macular degeneration. AB - PURPOSE: To compare 1-year safety and efficacy of anecortave acetate 15 mg with photodynamic therapy (PDT) with verteporfin in patients eligible for initial PDT treatment. DESIGN: Prospective, masked, randomized, multicenter, parallel group, active control, noninferiority clinical trial. PARTICIPANTS: Five hundred thirty patients with predominantly classic subfoveal choroidal neovascularization secondary to age-related macular degeneration were randomized to treatment with either anecortave acetate 15 mg or PDT. METHODS: In the anecortave acetate group, the drug was administered under the Tenon's capsule as a periocular posterior juxtascleral depot (PJD) at the beginning of the study and at month 6. Before the first administration of anecortave acetate, patients in this treatment group received a sham PDT treatment, and sham PDT treatments were repeated every 3 months if there was evidence of leakage on fluorescein angiography (FA). Patients assigned to PDT received up to 4 PDT treatments at 3-month intervals, as needed based upon FA, and a sham PJD procedure at the beginning of the study and at month 6. Best-corrected visual acuity was determined at baseline and all follow up visits. Safety data were regularly reviewed by an independent safety committee. MAIN OUTCOME MEASURE: Percent responders (patients losing <3 lines of vision) at month 12. RESULTS: Percent responders in the anecortave acetate and PDT groups were 45% and 49%, respectively (not statistically different, P = 0.43). The confidence interval (CI) for the difference ranged from -13.2% favoring PDT to +5.6% favoring anecortave acetate. The month 12 clinical outcome for anecortave acetate was improved in patients for whom reflux was controlled and who were treated within the 6-month treatment window (57% vs. 49%; 95% CI, 4.3% favoring PDT to +21.7% favoring anecortave acetate). No serious adverse events related to the study drug were reported in either treatment group. CONCLUSIONS: The safety and efficacy outcomes in this study demonstrate that the benefits of anecortave acetate for the treatment of choroidal neovascularization outweigh the risks associated with either the drug or the PJD administration procedure. PMID- 16368147 TI - Is there an association between diabetes and keratoconus? AB - PURPOSE: To examine whether there is an association between diabetes mellitus (DM) and keratoconus. DESIGN: A retrospective comparison of the proportion of keratoconus patients with DM versus the proportion of overall patients with DM, and a retrospective cross-sectional study of a cohort consisting of all diabetic keratoconus patients and randomly selected keratoconus patients without DM at a single center. PARTICIPANTS: Patients seen at the Wilmer Eye Institute from January 1, 1995, through March 18, 2004. METHODS: Review of billing data and clinic charts. Eligibility criteria for the cross-sectional study were 1 or more clinic visits, lack of other eye pathologic features (excluding cataract and diabetic retinopathy), and absence of bilateral penetrating keratoplasty (PK) at presentation. Application of novel keratoconus severity index was based on best corrected visual acuity (BCVA) in the better eye at last visit and defined as: grade 1 (least severe), spectacle wear with BCVA of 20/40 or better; grade 2 (intermediate), spectacle wear with BCVA worse than 20/40 or rigid gas permeable lens wear; grade 3 (most severe), PK. MAIN OUTCOME MEASURES: Prevalence of DM in keratoconus patients and those without keratoconus, odds ratio of having DM on a diagnosis of more severe keratoconus, and prevalence of DM in keratoconus patients and those without keratoconus who underwent corneal transplantation. RESULTS: There was no difference in the prevalence of DM in keratoconus patients and those without keratoconus, and there was no difference in the prevalence of DM in keratoconus patients and those without keratoconus undergoing PK. However, our results suggest a negative association between DM and severity of keratoconus (P = 0.03, Fisher exact test). The odds of being in the most severe group as opposed to the least severe group were lower in DM patients than in those without DM (P = 0.01; odds ratio [OR] = 0.20; 95% confidence interval [CI], 0.05-0.70). Compared with those without DM, DM patients also had lower odds of being in the intermediate group than in the least severe group (P = 0.02; OR = 0.25; 95% CI, 0.08-0.80). After adjustment for age, gender, and race, these differences remained statistically significant. CONCLUSIONS: We found that DM is not associated with a diagnosis of keratoconus, but having DM decreases the odds of having more severe keratoconus. PMID- 16368149 TI - The involvement of selectins and their ligands in tumor-progression. AB - About 70 years ago, Peyton Rous described the progression of cancer towards metastasis formation as "the process whereby tumors go from bad to worse". The interactions of tumor cells with endothelium are pivotal steps in this process. This review focuses on the role played by the selectins and their ligands in these interactions and especially in tumor cell extravasation. The working hypothesis of researchers studying tumor cell extravasation is that the tumor cells follow the extravasation strategy of leukocytes in their migration towards inflammatory sites. A significant portion of this review is, therefore, dedicated to the molecular mechanisms underlying leukocyte extravasation and to a comparison between the extravasation strategy employed by leukocytes and tumor cells. The review also summarizes some of the available data on signals generated by selectin-selectin ligand interactions. PMID- 16368148 TI - Neuroactive steroid effects on cognitive functions with a focus on the serotonin and GABA systems. AB - This article will review neuroactive steroid effects on serotonin and GABA systems, along with the subsequent effects on cognitive functions. Neurosteroids (such as estrogen, progesterone, and allopregnanolone) are synthesized in the central and peripheral nervous system, in addition to other tissues. They are involved in the regulation of mood and memory, in premenstrual syndrome, and mood changes related to hormone replacement therapy, as well as postnatal and major depression, anxiety disorders, and Alzheimer's disease. Estrogen and progesterone have their respective hormone receptors, whereas allopregnanolone acts via the GABA(A) receptor. The action of estrogen and progesterone can be direct genomic, indirect genomic, or non-genomic, also influencing several neurotransmitter systems, such as the serotonin and GABA systems. Estrogen alone, or in combination with antidepressant drugs affecting the serotonin system, has been related to improved mood and well being. In contrast, progesterone can have negative effects on mood and memory. Estrogen alone, or in combination with progesterone, affects the brain serotonin system differently in different parts of the brain, which can at least partly explain the opposite effects on mood of those hormones. Many of the progesterone effects in the brain are mediated by its metabolite allopregnanolone. Allopregnanolone, by changing GABA(A) receptor expression or sensitivity, is involved in premenstrual mood changes; and it also induces cognitive deficits, such as spatial-learning impairment. We have shown that the 3beta-hydroxypregnane steroid UC1011 can inhibit allopregnanolone induced learning impairment and chloride uptake potentiation in vitro and in vivo. It would be important to find a substance that antagonizes allopregnanolone induced adverse effects. PMID- 16368150 TI - Interleukin-18 induces the production of vascular endothelial growth factor (VEGF) in rheumatoid arthritis synovial fibroblasts via AP-1-dependent pathways. AB - Interleukin-18 (IL-18) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis in rheumatoid synoviocytes. In present study, we examined the effect of IL-18 on VEGF production in fibroblast-like synoviocytes (FLS) isolated from the patients with RA. FLS were prepared from the synovial tissues of patients with RA and osteoarthritis (OA) and cultured in the presence of IL-18. The production of VEGF from FLS was measured in culture supernatants by enzyme-linked immunosorbent assay (ELISA). The VEGF messenger RNA (mRNA) expression and AP-1 binding activity of VEGF transcript were determined by reverse transcription-polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assay (EMSA). IL-18 and VEGF levels of sera and synovial fluids (SF) of RA patients (n=30) were significantly higher than those of OA patients (n=20). IL-18 dose-dependently increased the production of VEGF. The effect of IL-18 on VEGF production appeared to be as potent as IL-1beta, whereas tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma showed little effects on VEGF production. AP-1-specific inhibitor Curcumin dose-dependently abrogated the effect of IL-18 on VEGF production. The VEGF enhancement of IL-18 was associated with increased AP-1 binding activity to the VEGF promoter site. These findings suggest IL-18 as an angiogenic factor in RA and down-regulation of IL-18 activity or AP-1 signal pathway can be potential therapeutic targets for RA. PMID- 16368151 TI - Still waiting for the toll? AB - Multicellular organisms including invertebrates and vertebrates live in various habitats that may be aquatic or terrestrial where they are constantly exposed to deleterious pathogens. These include viruses, bacteria, fungi, and parasites. They have evolved various immunodefense mechanisms that may protect them from infection by these microorganisms. These include cellular and humoral responses and the level of differentiation of the response parallels the evolutionary development of the species. The first line of innate immunity in earthworms is the body wall that prevents the entrance of microbes into the coelomic cavity that contains fluid in which there are numerous leukocyte effectors of immune responses. When this first barrier is broken, a series of host responses is set into motion activating the leukocytes and the coelomic fluid. The responses are classified as innate, natural, non-specific, non-anticipatory, non-clonal (germ line) in contrast to the vertebrate capacity that is considered adaptive, induced, specific, anticipatory and clonal (somatic). Specific memory is associated with the vertebrate response and there is information that the innate response of invertebrates may under certain conditions possess specific memory. The invertebrate system when challenged affects phagocytosis, encapsulation, agglutination, opsonization, clotting and lysis. At least two major leukocytes, small and large mediate lytic reactions against several tumor cell targets. Destruction of tumor cells in vitro shows that phagocytosis and natural killer cell responses are distinct properties of these leukocytes. This has prompted newer searches for immune function and regulation in other systems. The innate immune system of the earthworm has been analyzed for more than 40 years with every aspect examined. However, there are no known entire sequences of the earthworm as exists in these other invertebrates. Because the earthworm lives in soil and has been utilized as a successful monitor for pollution, there are studies that reveal up and down regulation of responses in the immune system after exposure to a variety of environmental pollutants. Moreover, there are partial sequences that appear in earthworms after exposure to environmental pollutants such as cadmium and copper. There are now attempts to define the AHR receptor crucial for intracellular signaling after exposure to pollutants, but without linking the signals to changes in the immune system. There are several pathways for signal transduction, including JAK/STAT, TOLL, TRAF PIP3, known in invertebrates and vertebrates. For resistance to pathogens, conserved signal transduction components are required and these include a Toll/IL-1 receptor domain adaptor protein that functions upstream of a conserved p38 MAP kinase pathway. This pathway may be an ancestral innate immune signaling pathway found in a putative common ancestor of nematodes, arthropods and even vertebrates. It could also help us to link pollution, innate immunity and transduction in earthworms. PMID- 16368152 TI - The effect of Vitamin E treatment on the development of myringosclerosis after ventilation tube insertion. AB - OBJECTIVES: Recent studies have established the relationship between the reactive oxygen species and myringosclerosis. Furthermore several antioxidants have been known to prevent myringosclerosis. All the previous studies supporting this hypothesis have been performed on animals. The aim of our study is to investigate the possible effect of Vitamin E on the development of tympanosclerosis after VT insertion on human subjects. METHODS: 72 children undergoing myringotomy and VT insertion were included in the study. Vitamin E was applied to the right ear and no treatment was applied to the left ear. Both ears were examined at the end of 9 months with otomicroscopy. Myringosclerosis formation at the end of the study period was noted for each ear. RESULTS: Myringosclerosis was found in 33 of the 144 ears. The overall incidence was 22.9%. While 22 (30.6%) of the 72 left ears showed myringosclerosis otomicroscopically, in only 11 (15.3%) of the 72 right ears that were treated with Vitamin E was myringosclerosis observed at the end of the study period. Of these nine cases were bilateral. CONCLUSION: Animal studies have well documented the development of myringosclerosis after myringotomy and VT insertion and beneficial effects of different antioxidants. Our study has shown similar results in human subjects. Further clinical studies consisting of a larger patient population are needed to bring about routine clinical use of antioxidants in myringotomy and VT insertion. PMID- 16368153 TI - A novel real-time PCR assay to determine relative replication capacity for HIV-1 protease variants and/or reverse transcriptase variants. AB - The emergence of drug-resistant viruses is a major issue in the treatment of HIV 1 infections. Quite often these drug-resistant viruses have a reduced replication capacity. A novel assay was developed to study the impact of mutations selected during therapy on viral replication capacity. Two HIV-1 HXB2 reference clones were constructed for this assay based on viral competition experiments, which are identical except for the presence of two silent nucleotide changes in p24 in one of the two clones. Within these two reference clones, three different contiguous deletions were constructed: (I) the C-terminus of Gag and protease, (II) the N terminus of RT and (III) the C-terminus of Gag and protease together with the N terminus of RT. Using these reference clones, recombinant viruses were created and viral competition experiments were performed. The proportion of each virus during the competition experiments was determined with a real-time PCR assay based on the two silent nucleotide changes in p24 in one of the two reference clones. With this novel assay it was possible to detect accurately differences in replication capacity due to mutations in the C-terminus of Gag and protease and/or the N-terminus of RT. PMID- 16368154 TI - Chemokine scavenging by the human cytomegalovirus chemokine decoy receptor US28 does not inhibit monocyte adherence to activated endothelium. AB - The human cytomegalovirus has found smart ways to exploit the chemokine network in order to subvert immune attack. Chemokines trigger the arrest and firm adhesion of inflammatory cells to the vascular wall. Scavenging of chemokines by viral decoy receptors, such as US28, might prevent arrest of leukocytes to the vascular wall and impair an antiviral immune response. We determined the effect of chemokine scavenging by endothelium-expressed signaling mute US28 (US28R129A) on static monocyte adhesion. Despite the chemokine scavenging capacity of US28R129A, expression of this construct by endothelial cells was insufficient to disrupt leukocyte adhesion to cytokine-activated monolayers. Our results suggest that the concentrations of chemokines that trigger firm leukocyte adhesion are too high to be efficiently scavenged by viral chemokine decoy receptors like US28. From the results of this experimental model a role for US28 in viral immune evasion by chemokine scavenging would appear therefore unlikely. PMID- 16368155 TI - The pleasurable recreational activities among community-dwelling older adults. AB - This study aimed to clarify what pleasurable recreational activities older adults like to participate in, and to investigate the relationship between those activities and quality of life (QOL). Questionnaires were delivered to older residents (65 years and above) in a Japanese rural area. The residents' background information, the amount of pleasure for various activities, and the QOL were surveyed. The QOL was evaluated by the revised Philadelphia Geriatric Center (PGC) morale scale. The amount of pleasure taken in a majority of the activities, such as conversation with family or neighbors showed a significant association with the happiness score, but only a few activities showed significant association between the revised PGC morale scale and the amount of pleasure. The multiple regression analyses indicated that the amount of pleasure in exercise, the difficulty in managing finances, and amount of pleasure taken in watching TV were significant variables for predicting the happiness score. The results indicated that the amount of pleasure older adults experienced when engaging in activities such as conversation with family or neighbors showed significant association with the older adults' happiness. These results may be helpful in understanding contributions of various activities to the perception of pleasure in older adults. PMID- 16368156 TI - Improved detection of acute myocardial infarction using a diagnostic algorithm based on calculated epicardial potentials. AB - BACKGROUND: New methods for detecting myocardial infarction in patients with suspected acute coronary syndromes are needed particularly in an era where the majority of patients with myocardial infarction present with non-diagnostic 12 lead electrocardiograms (ECG). We compared a novel epicardial diagnostic algorithm using epicardial potentials from the 80-lead body surface map with other electrocardiographic techniques in detection of myocardial infarction. METHODS: Between February 1999 and February 2001, consecutive patients (n=427) with ischemic type chest pain had an initial 12-lead ECG and body surface map recorded. Detecting myocardial infarction using an epicardial algorithm was first performed in a training set (n=213) and tested in a validation set of patients (n=214). The results from this epicardial algorithm in myocardial infarction detection were compared with the physician's interpretation of the 12-lead ECG, the body surface map algorithm (PRIME) and physician's interpretation of the body surface map. RESULTS: Myocardial infarction occurred in 205 patients (creatine kinase >or=2x upper limit of normal with creatine kinase-MB >or=7% CK). The physician's interpretation of the 12-lead ECG identified 122 with myocardial infarction (sensitivity 60%, specificity 99%), the body surface map algorithm 137 (sensitivity 67%, specificity 89%), the physician's interpretation of the body surface map 153 (sensitivity 75%, specificity 91%) and the epicardial algorithm 158 (sensitivity 77% specificity 99%). Combining the physician's interpretation of the 12-lead ECG with the epicardial algorithm increased significantly the detection of myocardial infarction (sensitivity 85%, specificity 98%, p<0.001) compared with the 12-lead ECG. CONCLUSIONS: An epicardial algorithm based on epicardial potentials increases significantly the detection of myocardial infarction particularly among those with non-diagnostic 12-lead ECG's. PMID- 16368157 TI - Systemic antiviral silencing in plants. AB - RNA silencing controls numerous developmental processes in eukaryotic organisms from fungi, plants, to animals. In plants as well as in animals, this system of RNA regulation functions as part of an immune response against invading viruses. From transitive RNA silencing to virus-induced gene silencing (VIGS), the systemic effects are proven to be the core of RNA silencing. This article reviews the latest advances in view of the effect of cellular RDR6, an RNA-dependent RNA polymerase (RdRp), on systemic RNA silencing, systemic virus silencing, and discusses the abilities of viral suppressors in modulating RNA silencing efficiency to establish effective infection. PMID- 16368158 TI - Interaction of targeted liposomes with primary cultured hepatic stellate cells: Involvement of multiple receptor systems. AB - BACKGROUND/AIMS: In designing a versatile liposomal drug carrier to hepatic stellate cells (HSC), the interaction of mannose 6-phosphate human serum albumin (M6P-HSA) liposomes with cultured cells was studied. METHODS: M6P-HSA was covalently coupled to the liposomal surface and the uptake and binding of 3H labelled M6P-HSA liposomes by primary rat HSC and liver endothelial cells was determined. The targeting ability of M6P-HSA liposomes to HSC was tested in bile duct ligated rats using immunohistochemical methods. RESULTS: The association of M6P-HSA liposomes with HSC was 4-fold higher than of control liposomes. An excess of M6P-HSA inhibited this association by 58%, indicating M6P receptor specificity. The scavenger receptor competitor polyinosinic acid abolished association of M6P-HSA liposomes with HSC. M6P-HSA liposomes also amply associated with endothelial cells, which abundantly express scavenger receptors. Endocytosis of M6P-HSA liposomes by HSC was temperature dependent and could be inhibited by monensin. In the fibrotic liver M6P-HSA liposomes co-localised with HSC. CONCLUSIONS: Coupling of M6P-HSA to liposomes strongly increases the in vitro uptake of these liposomes by HSC and endothelial cells. Both the mannose 6 phosphate receptor and the scavenger receptors are involved in the uptake process. M6P-HSA liposomes are potential drug carriers to HSC in the fibrotic liver. PMID- 16368159 TI - Candidates for therapy: HBV. AB - Hepatitis B may cause liver damage ranging from mild chronic hepatitis to severe active hepatitis, cirrhosis and hepatocellular carcinoma. HIV and HBV co infection is more likely to lead to lower rates of HBeAg seroconversion, and higher HBV DNA concentrations. Immune restitution may lead to more severe hepatitis. The timing of acquisition of HBV versus HIV will have a bearing on considerations of treatment. Patients may have acquired HIV super-infection of chronic hepatitis B, HBV super-infection of HIV; alternatively, reactivation of hepatitis B may occur in a HIV positive patient, or the patient may be co infected at diagnosis. The patient may be naive or experienced or have resistant (HBV) at the time of superinfection. The risk of death is higher in patients with co-infection compared to those with HBV alone. The goals of therapy for hepatitis B are to prevent progression of the disease. If HBV replication can be suppressed, the accompanying reduction in histological activity lessens the risk of progression. Patients may request treatment to reduce infectivity, and this is relevant in co-infected patients. HBV has little effect on HIV or the effect of treatment on HIV; however, HIV, and HIV treatment profoundly affects the natural history of HBV. Therefore, it is usually important to target treatment of HBV to alter the outcome and take into account the impact of HBV treatment on HIV. Special concepts of treatment are applicable in HIV and HBV co-infected patients. PMID- 16368160 TI - Plant-microbe interactions to probe regulation of plant carbon metabolism. AB - Plant growth and development is dependent on coordinated assimilate production, distribution and allocation. Application of biochemical and molecular techniques substantially contributed to a better understanding of these processes, although the underlying regulatory mechanisms are still not fully elucidated and attempts to improve crop yield by modulating carbon partitioning were only partially successful. Plant pathogens also interfere with source-sink interaction. To this end they have evolved a wide range of sophisticated strategies to allow their systemic spread, suppression of plant defence and induction of sink function to support nutrient acquisition for their growth. Studying compatible interactions of plants and pathogens like viruses, bacteria and fungi can be exploited to investigate different levels of source-sink regulation. The identification of microbial factors and their host targets involved in regulation of plant primary metabolism may allow developing novel strategies to increase crop yield. Here we will discuss recent studies on plant-microbe interactions aimed at elucidating mechanisms of compatibility. PMID- 16368161 TI - Role of asparagine and asparagine synthetase genes in sunflower (Helianthus annuus) germination and natural senescence. AB - Sunflower (Helianthus annuus) contains three active asparagine synthetase (EC 6.3.5.4, AS) genes: HAS1, HAS1.1 and HAS2. Asparagine content and AS gene expression were determined during germination and leaf and cotyledon natural senescence to assess the role of asparagine as well as the extent of participation of each AS gene in different nitrogen mobilizing processes. Asparagine accumulated in the dry seed and was the predominant amide throughout germination. During cotyledon senescence, the asparagine level was slightly higher than that of glutamine. The opposite was true for leaf senescence. According to transcript accumulation data, most of the asparagine newly synthesized for germination and cotyledon expansion was due to HAS2 activity, with little contribution of the other AS genes. However, all three genes work together to synthesize asparagine for leaf senescence. The absence of significant AS gene expression in cotyledon senescence differentiates leaf and cotyledon senescence, and suggests a cotyledon-specific regulation. PMID- 16368162 TI - The wound response in tomato--role of jasmonic acid. AB - Plants respond to mechanical wounding or herbivore attack with a complex scenario of sequential, antagonistic or synergistic action of different signals leading to defense gene expression. Tomato plants were used as a model system since the peptide systemin and the lipid-derived jasmonic acid (JA) were recognized as essential signals in wound-induced gene expression. In this review recent data are discussed with emphasis on wound-signaling in tomato. The following aspects are covered: (i) systemin signaling, (ii) JA biosynthesis and action, (iii) orchestration of various signals such as JA, H2O2, NO, and salicylate, (iv) local and systemic response, and (v) amplification in wound signaling. The common occurrence of JA biosynthesis and systemin generation in the vascular bundles suggest JA as the systemic signal. Grafting experiments with JA-deficient, JA insensitive and systemin-insensitive mutants strongly support this assumption. PMID- 16368163 TI - Medicinal chemistry tools: making sense of HTS data. AB - The main problem in QSAR modeling from the high throughput screening (HTS) data is that by definition, it produces only a small proportion of hits against a given assay. This leads to a very small statistical significance of the hits in comparison with the "noise". Analysis based purely on the "hit" compounds removes useful information about the biological response of all the test compounds. What is needed is an analysis technique that increases the significance of the active compounds, while using the information present in the original data. In this paper we present a method for application of intelligent filtering of the data to improve statistical significance of the active compounds to generate predictive models that provide medicinal chemists with a powerful tool for both optimizing compounds and mining screening candidates in libraries. PMID- 16368164 TI - Synthesis and biological properties of new alpha-methylene-gamma-butyrolactones and alpha,beta-unsaturated delta-lactones. AB - The synthesis of a series of alpha-methylene-gamma-butyrolactones (compounds 4a, 4b, 6-12, 16, 17) and alpha,beta-unsaturated-delta-lactones (compounds 19-23, 25, 26) starting from 4,4-dimethyldihydrofuran-2,3-dione (1) has been described. Their chemical structures were assigned by spectroscopic evidence. These new compounds exhibited significantly different antiproliferative properties against cultured human tumor cell lines with their IC(50) values ranging from 0.88 to>20.00 microM. PMID- 16368165 TI - [NK cells: new insights on physiology and clinical implication in diseases]. AB - INTRODUCTION: Natural killer cells are cytotoxic lymphocytes of innate immunity. These last ten years our knowledge about the mechanisms that regulates NK cell function has greatly improved. Our purpose is to present a review of these new acquisitions and their potential implications in human disease. CURRENT KNOWLEDGE AND KEY POINTS: NK cell function is regulated by a repertoire of NK cell receptors and is diversified by recognition of MHC class I by a multigenic and multi-allelic family of NK receptors. Analysis of NK cell repertoire has been used to investigate features that characterize NK cells in pathological situations. Apart from their direct cytotoxic potential to eliminate target cells, recently identification of mechanisms that control NK cell mediated cytokine production and cross talk with dendritic cells emphasize the role of NK cells in the regulation of acquired immune response. FUTURE PROSPECTS AND PROJECTS: These findings have lead to a better knowledge of the importance of the NK cells in several human diseases. It has been shown that NK cells are actors of the immunosurveillance of tumoral and infectious challenges. Allo or auto reactivity of the NK cell compartment have also been suggested in autoimmune diseases, infertility or foetal loss and transplantation. Ongoing research on NK cells in the fields of human diseases is increasing and will clarify the utility of the evaluation of the NK cell compartment and their receptors in clinical practice. PMID- 16368166 TI - [Calciphylaxis: a rare differential diagnosis of systemic vasculitis]. AB - INTRODUCTION: Calciphylaxis is a rare phenomenon of medium- and small-vessel calcifications leading to cutaneous necrosis mimicking vasculitis. CASE REPORT: A 75 year-old-woman with chronic renal insufficiency was admitted for extensive cutaneous necrosis of the limb. Diagnosis of vasculitis was made, but inspite of corticosteroid therapy, the condition of the patient was worsening. After cutaneous biopsy, the diagnosis of calciphylaxis was established. CONCLUSION: Calciphylaxis must be suspected in cases of cutaneous necrosis occurring in a patient with chronic renal failure. Treatment requires rapid normalization of phosphocalcic balance. It is a condition with high mortality. PMID- 16368168 TI - A novel strategy to compensate the disadvantages of live vaccine using suicide gene system and provide better antitumor immunity. AB - Fusing dendritic cells (DCs) with tumor cells is a powerful vaccine to increase tumor immunogenicity. To develop more effective and safer therapeutic vaccine, we fused rat bone marrow-derived DCs with ovarian tumor cell line NuTu-19 modified by suicide gene (HSV1-TK gene) to obtain live vaccine against ovarian cancer. Our data showed that immunization of rats with such live vaccine solicited stronger ovarian tumor-specific cytotoxic T lymphocyte responses and induced immunopreventive and immunotherapeutic effects against parental tumor cells in vivo. Live vaccine could be induced to death after ganciclovir administration in vitro and in vivo. Our researches suggest that live vaccine modified with suicide gene might be effective and controllable in the therapy of ovarian cancer. PMID- 16368169 TI - Diphtheria tetanus pertussis poliomyelitis vaccination and reported atopic disorders in 8-12-year-old children. AB - BACKGROUND: Evidence for the relationship between the diphtheria tetanus pertussis (DTP) vaccination and atopic disorders is inconclusive, because the available studies that constitute the evidence are liable to confounding by indication. STUDY OBJECTIVE: To assess the relationship between diphtheria tetanus-pertussis-(inactivated) poliomyelitis vaccination (DTP-IPV) in the first year of life and reported atopic disorders at primary school age. METHOD: 1875 children attending Orthodox Reformed (Protestant) primary schools in the Netherlands returned questionnaires with data on vaccination status, atopic symptoms and lifetime atopic disorders (asthma, hay fever, eczema and food allergy), and possible confounders. RESULTS: The adjusted odds ratio of any atopic disorder (vaccinated/unvaccinated) was 1.00 (CI95%: 0.80-1.24). For asthma, hay fever, eczema and food allergy the results were respectively: 1.04 (CI95%: 0.76-1.42), 0.79 (CI95%: 0.55-1.12), 0.87 (CI95%: 0.66-1.14) and 1.13 (CI95%: 0.71-1.81). CONCLUSION: The DTP-IPV vaccination was not related to reported atopic disorders at primary school age. PMID- 16368167 TI - Prototype Alzheimer's disease epitope vaccine induced strong Th2-type anti-Abeta antibody response with Alum to Quil A adjuvant switch. AB - Beta-amyloid (Abeta) peptide has been proposed to be a causal factor in Alzheimer's disease (AD). Currently being investigated, active and passive Abeta immunotherapy significantly reduce Abeta plaque deposition, neuritic dystrophy, and astrogliosis in the brains of APP transgenic (APP/Tg) mice. Immunization with Abeta42 formulated in the Th1-type adjuvant QS21 was beneficial for AD patients with significant titers of anti-Abeta antibodies, however, 6% of participants developed meningoencephalitis, likely due to anti-Abeta-specific autoimmune Th1 cells. Thus, successful Abeta vaccination requires the development of strong antibody responses without Th1-type cellular immunity. In this study, we compared the induction of humoral immune responses with Th1-type (Quil A) and Th2-type (Alum) adjuvants singly and in combination, using our novel epitope vaccine composed of self B cell epitope Abeta(1-15) and foreign T cell epitope PADRE (PADRE-Abeta(1-15)-MAP). Formulated in Quil A, this vaccine resulted in significantly higher anti-Abeta antibody responses in both BALB/c (H-2d) and C57BL/6 (H-2b) mice, compared with Alum. Anti-Abeta antibodies induced by Alum were predominantly IgG1 type accompanied by lower levels of IgG2a and IgG2b. Quil A induced robust and almost equal titers of anti-Abeta antibodies of IgG1 and IgG2a isotypes and slightly lower levels of IgG2b. Switching adjuvants from Alum to Quil A induced higher concentrations of antibodies than injections with Alum only, however slightly lower than Quil A only. Switching both adjuvants did not change the profile of antibody responses generated by the initial adjuvant injected. These results suggest that switching from Alum to Quil A would be beneficial for AD patients because anti-Abeta antibody production was enhanced without changing the initially generated and likely beneficial Th2-type humoral response. PMID- 16368170 TI - Disappearing condyles in a patient with seronegative spondylarthropathy. AB - Severe osteolysis of the temporomandibular joint is unusual and can be associated with systemic conditions. We describe a 21-year-old white woman with an unusual pattern of extensive resorption and regeneration of the mandibular condyles who was treated with methotrexate. PMID- 16368171 TI - Intrapartal care documented in a Swedish maternity unit and considered in relation to World Health Organization recommendations for care in normal birth. AB - OBJECTIVES: To describe documented intrapartal care in relation to the World Health Organization (WHO) recommendations for care in normal birth, and to compare intrapartal care for pregnant women at low and high risk in a conventional maternity unit. DESIGN: Retrospective examination of 212 consecutive childbirth records using an audit instrument developed from WHO's recommendations. SETTING: A conventional maternity unit in Western Sweden. FINDINGS: Practices that are demonstrably useful and should be encouraged were mostly documented, except for physical assessments, such as pulse and temperature and emotional aspects. Vaginal examinations were carried out more often than recommended, and fetal heart rates were seldom monitored intermittently. Practices classified as harmful, practices with insufficient evidence and practices frequently used inappropriately, were used to a large extent. There were high rates of interventions regardless of the women's risk level. The interventions were carried out without a rational documented indication. According to the documentation, only two-thirds of the women were in active labour on admission to the labour ward. CONCLUSION AND IMPLICATIONS FOR PRACTICE: The recommendations from WHO were only partly adhered to. The instrument is considered useful for systematic audit of documented intrapartal care, and may help to identify areas in need of improvement. Improvements suggested by this study were as follows: inclusion of emotional aspects in the documentation, differentiation in cardiotocographic (CTG) surveillance for women at low and high risk, documentation of explicit indications for interventions and guidelines for admission to the maternity unit. PMID- 16368172 TI - Immediate corticocancellous bone autografting in segmental bone defects of the hand. AB - We performed a retrospective analysis of 12 type III open hand fractures in seven patients with segmental bone loss and associated soft tissue injuries to determine the effectiveness of immediate autologous corticocancellous bone grafting. Radical debridement and fracture fixation were performed prior to bone grafting. Results were interpreted according to clinical and radiologic time of bony healing, rate of infection, time to return to regular work duty, grip strength, rate of complications and range of motion. The final union rate was 92%, with a mean time to bony union of 18 weeks. The infection rate was 0%. The mean time to return to regular work duty, including recovery time after secondary surgeries, was 5 months and 21 days. All patients returned to their pre-injury employment. The mean total active motion of the combined metacarpophalangeal proximal interphalangeal and distal interphalangeal joints in bone-grafted digits was 178 +/- 53 degrees at final follow-up. PMID- 16368173 TI - A review of hand surgery provision in England. AB - This study reviews hand surgical activity and the resources available for provision of hand surgery in England in 2001. Operation rates for three common procedures, viz. carpal tunnel release, Dupuytren's surgery and ganglion surgery, were considered. The local population and the number of hand surgeons in each NHS Hospital Trust were compared. We identified 275 consultant surgeons with an interest in hand surgery working in the NHS in England. Approximately two-thirds were orthopaedic surgeons, almost one-third were plastic surgeons and a small number were accident and emergency surgeons. Half of all hand surgeons worked in large units, with three or more hand surgeons, but almost 20% of hand surgery was delivered in hospitals in which there was no surgeon with a declared interest in hand surgery. Surgery rates for Dupuytren's contracture varied from 0.04 to 0.36 cases per 1,000 population per annum and for carpal tunnel syndrome varied from 0.25 to 1.31 cases per 1,000 per annum. We found a correlation between rates of surgery and the number of hand surgeons, locally. A recent audit (Burke, Dias, Heras-Pelou, Bradley, & Wildin, 2004. Providing care for hand disorders, a reappraisal of need. Journal of Hand Surgery, 29B: 575-579.) has suggested that one hand surgeon is required to meet the needs of a population of 125,000, with a national requirement for 393 hand surgeons. We conclude that there are insufficient hand surgeons in England and believe that the wide local variations in hand surgery rates are indicative of a significant unmet demand for hand surgery in the English population. PMID- 16368174 TI - Biomonitoring using the lichen Hypogymnia physodes and bark samples near Zlatna, Romania immediately following closure of a copper ore-processing plant. AB - Lichen transplants were established along a 40km transect centred on a large mine waste dump close to Zlatna town centre, two weeks after closure of a major industrial source, to compare spatial patterns of element concentrations in lichen and bark samples. After 3 months of exposure, spatial patterns of 4 element concentrations (Pb, Cu, Zn and Fe) in transplants confirmed deposition due to a point source 'Zlatna influence'. Cu and Pb reached concentrations 8 and 4 times, respectively, higher than samples transplanted in the 'background' site, and over 200 (Cu) and 2000 (Pb) times higher relative to 'background' bark. Ten out of 15 elements analysed reached highest concentrations in bark. Spatial patterns confirmed long-term pollution for 6 elements (Pb, S, Fe, Cu, Zn and Ba). The study provides a model baseline to monitor recovery following closure of major industrial sources and highlights the importance of considering biogeochemical processes when interpreting metal concentrations. PMID- 16368175 TI - Increased formation of carcinogenic PAH metabolites in fish promoted by nitrite. AB - Nitrite (NO(2)(-)), a highly reactive chemical species, accumulates in coastal waters as a result of pollution with nitrogenous waste and/or an imbalance in the bacterial processes of nitrification and denitrification. The present study probed the impact of nitrite (NO(2)(-)) on the metabolism of polycyclic aromatic hydrocarbons (PAHs) in fish. In a laboratory experiment, exposure of euryhaline fish, Oreochromis mossambicus to industrial effluents containing PAHs in the presence of NO(2)(-) enhanced the cytochrome P450-dependent biotransformation activity determined as 7-ethoxyresorufin-O-deethylase (EROD), by nearly 36% compared to the value observed in the absence of NO(2)(-) (50.2 +/- 6.74 pmol resorufin min(-1) g(-1) liver). Fixed wavelength fluorescence measurements in bile revealed maximum enhancement to have occurred in the metabolites of benzo[a]pyrene, a carcinogenic PAH. Lasting, sublethal physiological deterioration was apparent in fish exposed simultaneously to an oil refinery effluent and NO(2)(-), from the unremittingly decreasing liver somatic index, even after the withdrawal of the contaminants. PMID- 16368176 TI - Remediation efficiency of vapour extraction of sandy soils contaminated with cyclohexane: Influence of air flow rate, water and natural organic matter content. AB - This work reports the analysis of the efficiency and time of soil remediation using vapour extraction as well as provides comparison of results using both, prepared and real soils. The main objectives were: (i) to analyse the efficiency and time of remediation according to the water and natural organic matter content of the soil; and (ii) to assess if a previous study, performed using prepared soils, could help to preview the process viability in real conditions. For sandy soils with negligible clay content, artificially contaminated with cyclohexane before vapour extraction, it was concluded that (i) the increase of soil water content and mainly of natural organic matter content influenced negatively the remediation process, making it less efficient, more time consuming, and consequently more expensive; and (ii) a previous study using prepared soils of similar characteristics has proven helpful for previewing the process viability in real conditions. PMID- 16368177 TI - The Akoya pearl oyster shell as an archival monitor of lead exposure. AB - The Akoya pearl oyster (Pinctada imbricata) was experimentally exposed to (a) constant levels of lead (Pb) at 180 microg L(-1) for nine weeks, or (b) two short term (pulse) exposures of Pb at 180 microg L(-1) (three weeks each) with an intervening depuration period (three weeks), to assess its utility as an (i) accumulative monitor of Pb contamination and an (ii) archival monitor for discriminating constant versus pulsed Pb exposure events. P. imbricata showed similar reductions in growth (based on shell morphology and wet weight) and Pb accumulation patterns for whole tissue and shell in response to both Pb exposure regimes. Thus the whole oyster was deemed an inappropriate accumulative monitor for assessing short-term temporal variation of Pb exposure and effect. However, using secondary ion mass spectrometry, Pb was shown to accumulate in the successively deposited nacreous layers of the shell of P. imbricata, documenting the exposure history of constant versus pulsed Pb events. Patterns of Pb deposition not only reflected the frequency of Pb exposure events but also their relative durations. Thus, the shell of P. imbricata may be employed as a suitable biological archive of Pb exposure. PMID- 16368178 TI - Race, socioeconomic status, and the perceived importance of positive self presentation in health care. AB - Hundreds of studies have documented disparities in medical treatment in the USA. These findings have generated research and initiatives intended to understand and ameliorate such disparities. Many articles examine disadvantaged patients' beliefs and attitudes toward health care, but generally limit their investigation to how these beliefs and attitudes influence adherence and utilization. Thus, this approach fails to consider whether patients use particular strategies to overcome providers' potentially negative perceptions of them and/or obtain quality medical care. In this paper, we examine positive self-presentation as a strategy that may be used by disadvantaged groups to improve their medical treatment. Analysis of survey data (the 2004 Greater Cincinnati Survey) suggests that both African Americans and lower socioeconomic status persons are more likely than whites or higher socioeconomic status persons to report that positive self-presentation is important for their getting the best medical care. Based on these findings, we suggest several routes for future research that will advance our understanding of patients' everyday strategies for getting the best health care. PMID- 16368179 TI - Molecular mechanism investigation of cycloheximide-induced hepatocyte apoptosis in rat livers by morphological and microarray analysis. AB - Male F344 rats were intravenously treated with 6 mg/kg cycloheximide (CHX), and microarray analysis was conducted on their livers 1, 2 and 6h after the CHX treatment. The histopathological examination and serum chemistry results indicated a mild hepatic cell death 2 and 6h after the CHX treatment, respectively. Multi-focal hepatocellular necrosis with slight neutrophil infiltration was observed 6h after the CHX treatment. The TUNEL staining results showed that the number of apoptotic hepatocytes was the highest 2h after the CHX treatment. Dramatic increases in the mRNA levels of ATF3 and CHOP genes, both of which were reported to play roles in the ER stress-mediated apoptosis pathway, were observed from 1h after the CHX treatment. In addition, increase of GADD45, p21 and p53 mRNA levels also suggested a time course-related stimulation of hepatocellular apoptotic signals. These results suggest that the hepatocyte apoptosis induced by the CHX treatment is triggered by ER stress. The hepatic mRNA levels of proinflammatory genes, such as TNFalpha, IL-1alpha and beta, were also increased 1 and 2h after the CHX treatment, supposedly mediated by the activated Kupffer cells engulfing the apoptotic hepatocytes. PMID- 16368180 TI - Role of metal ions in ligand-receptor interaction: insights from structural studies. AB - Experimental data indicate that metal ions such as Na(+), Ca(2+) and Mg(2+), which are present in millimolar concentrations in the extracellular environment, modulate binding of ligands to plasma membrane receptors. Here, we briefly review structural studies that demonstrate that various types of ligands, including peptide hormones and drugs, bind metal ions, in particular Ca(2+), in the lipid milieu. We propose that the metal ion-bound forms of ligands represent their bioactive conformations. With a view to understanding the mechanism of modulation of ligand-receptor interactions by metal ions, we have computed a homology model of the mu-opioid receptor, a G protein-coupled receptor (GPCR), and performed docking of specific agonist and antagonist ligands in the receptor. This resulted in the formation of a ligand-metal ion-receptor (ternary) complex which accounted for the data on the structure-activity relationships of ligands and mutation data on the receptor. Based on experimental and modeling data, we have proposed a general mechanism of activation of GPCRs by their corresponding ligands wherein metal ions play a pivotal role. Studies on overexpressed segments of mu-receptor are in progress to verify the above proposal. PMID- 16368181 TI - Oestrogen-mediated tyrosine phosphorylation of caveolin-1 and its effect on the oestrogen receptor localisation: an in vivo study. AB - Recently, it has been shown that 17beta estradiol (E2) induces a rapid and transient activation of the Src ERK phosphorylation cascade: a clear indication that the alpha oestrogen receptor (ERalpha) is able to associate with the plasma membrane. Increasing evidence suggests that caveolae, which are caveolin-1 containing, highly hydrophobic membrane domains, play an important role in E2 induced signal transduction. Caveolae can accumulate signalling molecules preferentially; thus, they may have a regulatory role in signalling processes. Results from previous experiments have shown that E2 treatment decreased the number of surface connected caveolae significantly in uterine smooth muscle cells and also downregulated the expression of caveolin-1. In addition to providing further evidence that ERalpha interacts with caveolin/caveolae in uterine smooth muscle cells, this study also shows that the interaction between caveolin-1 and ERalpha is actually facilitated by E2. One of the signal transduction components found to accumulate in caveolae is Src kinase in an amount that increases simultaneously with increases in the amount of ERalpha. Upon E2 treatment, Src kinase is tyrosine phosphorylated, which, in turn, stimulates Src kinase to phosphorylate caveolin-1. Phosphorylation of caveolin-1 can drive caveolae to pinch off from the plasma membrane, thereby decreasing the amount of plasma membrane-associated caveolin-1. This loss of caveolin/caveolae activates the signal cascade that triggers cell proliferation. PMID- 16368182 TI - The ubiquitin-specific protease USP10 modulates androgen receptor function. AB - The role of the ubiquitin/proteasome system in degrading nuclear hormone receptors and regulating their transcriptional function has emerged in the last few years. We identified the ubiquitin-specific protease USP10 as part of DNA bound androgen receptor (AR) complexes purified from nuclear extracts of PC-3 cells stably expressing the AR. The interaction between USP10 and the AR was confirmed by GST pull-down assays. Fluorescence microscopy documented that USP10 was localised in the nucleus and the cytoplasm. Cell-based transactivation assays in PC-3/AR cells revealed that overexpression of wild-type USP10, but not of an enzymatically inactive form, stimulated AR activity mediated by reporter constructs harbouring selective androgen response elements (AREs), non-selective steroid response elements (SREs) or the mouse mammary tumour virus (MMTV) promoter. Conversely, USP10 expression knock-down by siRNAs impaired the MMTV response to androgen. In summary, the data indicate that USP10 is a new cofactor that binds to the AR and stimulates the androgen response of target promoters. This finding underlines the role of the ubiquitin/proteasome system in modulating the AR function. PMID- 16368183 TI - A generalized numerical approach to rapid-equilibrium enzyme kinetics: application to 17beta-HSD. AB - A generalized numerical treatment of rapid-equilibrium enzyme kinetics is presented. This new approach relies on automatic computer derivation of the underlying mathematical model (a system of simultaneous nonlinear algebraic equations) from a symbolic representation of the reaction mechanism (a system of biochemical equations) provided by the researcher. The method allows experimental biochemists to analyze initial-rate enzyme kinetic data without having to use any mathematical equations. An illustrative example is based on the inhibition kinetics of 17beta-hydroxysteroid dehydrogenase type 5 by a class of natural compounds. A computer implementation of the new method, a newly modified software package DYNAFIT [Kuzmic, P., 1996. Program DYNAFIT for the analysis of enzyme kinetic data: application to HIV proteinase. Anal. Biochem. 237, 260-273], is freely available to all academic researchers. PMID- 16368184 TI - Central nervous system involvement in systemic connective tissue diseases. AB - Systemic connective tissue diseases can affect the brain, meninges, spinal cord, cranial and peripheral nerves. Different pathogenic mechanisms, particularly autoantibody or T-cell mediated lesions, appear to be involved. The neurological manifestations of the connective tissue diseases and their diagnostic possibilities including newer imaging techniques are reviewed. Early recognition of neurological abnormalities can help in the differential diagnosis and in defining the underlying disease in order to initiate treatment and prevent progression of lesion or cognitive function loss. PMID- 16368186 TI - Use of polytetrafluoroethylene dural substitute as adhesion preventive material during craniectomies. AB - OBJECTIVE: We describe the use of an artificial dural substitute in order to prevent peridural fibrosis in patients who underwent craniectomy and subsequent cranioplasty. PATIENTS AND METHODS: In our institution we performed decompressive craniectomy and subsequent cranioplasty, with autologous bone flap, in 52 patients (mean age 32 years) between 1998 and 2004. Most of the craniectomy procedures were performed as emergency, brain swelling relief, measure for various reasons. The standard decompressive craniectomy technique that we perform includes bone removal, meningoplasty and protection from future adhesion formations in 23 of the 52 patients by the use of polytetrafluoroethylene (ePTFE) dural substitute (Preclude((R))). The substitute was applied over dural anasynthesis and under the galea, during craniectomy. The development of adhesion formations between tissue layers was evaluated during reoperation for cranioplasty. RESULTS: During cranioplasty reoperation we observed that the polytetrafluoroethylene dural substitute succeeds in creating a controlled dissection plane, facilitating access to the epidural space, shortening the operation time by approximately 25% and diminishes the intraoperative blood loss by 37% as compared with the group of patients underwent standard cranioplasty. The difference is of statistical significance (p<0.005). As a result, the use of polytetrafluoroethylene dural substitute as adhesion preventive material could make the cranioplasty operation safer and more successful. CONCLUSION: Polytetrafluoroethylene dural substitute is able to prevent peridural scarring and is very efficient in facilitating cranioplasty in patients who underwent craniectomy regardless the indication. PMID- 16368187 TI - Mass spectrometric identification of serine hydrolase OVCA2 in the medulloblastoma cell line DAOY. AB - OVCA2 is a putative serine-hydrolase. Performing protein profiling in human tumour cell lines, OVCA2 was detected in DAOY medulloblastoma cells as a high abundance protein. The protein was unambiguously identified by 2D gel electrophoresis and MALDI-MS and MS/MS, its presence was confirmed by western blotting. Immunohistochemistry revealed expression in medulloblastoma and predominantly in oligodendrocytes. Computational approaches predicted functional motifs and domains, interaction with apoptosis-related protein BAG and 3D structure. In addition to the presence of OVCA2 in medulloblastoma, it was furthermore detectable in three out of 10 human tumour cell-lines as a high abundance protein probably suggesting a role in the tumour biology. PMID- 16368188 TI - Equilibrium studies for the sorption of zinc and copper from aqueous solutions using sugar beet pulp and fly ash. AB - In the present work, the abilities of native sugar beet pulp (SBP) and fly ash (FA) to remove copper (Cu(2+)) and zinc (Zn(2+)) ions from aqueous solutions were compared. The SBP and FA, an industrial by-product and solid waste of sugar industry, were used for the removal of copper and zinc from aqueous water. Batch adsorption experiments were performed in order to evaluate the removal efficiency of SBP and lignite-based FA. The effect of various operating variables, i.e. initial pH, adsorbent dose, initial metal ion concentration, and time on adsorption of copper and zinc onto the SBP and FA, has been studied. The sorption process was relatively fast and equilibrium was reached after about 60 min of contact. As much as 60-97% removal of copper and zinc for SBP and FA are possible in about 60 min, respectively, under the batch test conditions. Uptake showed a pH-dependent profile. The overall uptake for the SBP is at a maximum at pH 5.5 and gives up to 30.9 mg g(-1) for copper and at pH 6.0 and gives 35.6 mg g(-1) for zinc for SBP, which seems to be removed exclusively by ion exchange and physical sorption. Maximum adsorption of copper and zinc occurred 7.0 and 7.84 mg g(-1) at a pH value of 5.0 and 4.0 for FA, respectively. A dose of 8 g l(-1) of SBP and 8 g l(-1) FA were sufficient for the optimum removal of both the metal ions. The sorption data were represented by the Freundlich for SBP and the Langmuir and Freundlich for FA. The sorption data were better represented by the Langmuir isotherm than by the Freundlich one for FA in the adsorption of zinc ion, suggesting that the monolayer sorption, mainly due to ion exchange. The presence of low ionic strength or low concentration of Na and Cl ions does not have a significant effect on the adsorption of these metals by SBP and FA. The SBP and FA are shown to be effective metal adsorbents for these two metals. PMID- 16368189 TI - Site-specific phosphorylation of SCG10 in neuronal plasticity: role of Ser73 phosphorylation by N-methyl D-aspartic acid receptor activation in rat hippocampus. AB - Accumulated evidence suggests that actin and microtubule regulating proteins contribute to neuronal structural dynamics, which subsequently affect neuronal plasticity. SCG10 is a neuronal-specific stathmin protein with microtubule destabilizing activity that is affected by multiple phosphorylation, at least in vitro. SCG10 has four major phosphorylation sites: Ser50 and Ser97 targeted by protein kinase A (PKA), and Ser62 and Ser73 targeted by mitogen-activated protein kinase (MAPK). To explore the potential roles of site-specific phosphorylation in physiological models, we developed phosphorylation site-specific antibodies and examined the SCG10 status in primary cultured hippocampal neurons and tissues. Although SCG10 is concentrated in growth cones and the Golgi apparatus in primary cultured neurons, the phosphorylated form was also detected in both regions, suggesting that MT dynamics within the growth cone may be regulated by protein phosphorylation. In the adult hippocampus, an intense stimulus such as kainate treatment induced a rapid phosphorylation of Ser73 within 15 min that was sustained for at least 60 min. This response was mediated through the N-methyl D aspartic acid (NMDA) receptor and was ablated by the antagonist MK-801. The MAPK enzyme Erk2 was simultaneously activated along a similar time course to SCG10, suggesting that Erk2 may directly phosphorylate Ser73. These results demonstrate that changes in the phosphorylation status of SCG10 in vivo, dependent upon neural activity and/or plasticity, could affect the microtubule dynamics in neuronal dendrites. PMID- 16368190 TI - A functional subdivision of the circadian clock is revealed by differential effects of melatonin administration. AB - The biological clock of the suprachiasmatic nuclei drives numerous physiological and behavioural circadian rhythms. In this study, we addressed the question as to whether different components of the clock may control separately various circadian functions. Using the rat transpineal microdialysis tool, we analysed the effect of clock perturbation by exogenous melatonin injection on two hormonal clock outputs: pineal melatonin and adrenal corticosterone secretions. As already reported, a single melatonin injection at the light/dark transition induces a marked increase in the endogenous pineal melatonin peak for the two following days. In the same animals, by contrast, the amplitude of the corticosterone rhythm was not altered following melatonin injection. These data show that the melatonin injection does not display an overall effect on the circadian clock, but rather influences a subpopulation of melatonin-sensitive neurons involved, among other functions, in the circadian control of the pineal pathway. PMID- 16368191 TI - Picrotoxin-induced convulsions alters rat brain microsomal membrane structural properties. AB - Cerebral microsomal membrane properties were assessed in the chronic condition of generalized seizure induced by picrotoxin (PTX) in rats. PTX-induced seizures resulted in increased lysophosphatidyl glycerol, phosphatidylcholine and phosphatidic acid components, while phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol were significantly reduced by 19-73%. The cholesterol (CHL) content increased considerably by 25% without alteration in total phospholipids content. Microsomal membrane was more fluidized in the epileptic condition. Possible consequences of microsomal membrane alterations are discussed in terms of deregulation of Ca2+ homeostasis. In conclusion, alterations in the microsomal membrane properties may have a significant influence on the cerebral function in the chronic epileptic condition. PMID- 16368192 TI - A PET activation study of brush-evoked allodynia in patients with nerve injury pain. AB - Acute experimental brush-evoked allodynia induces a cortical activation pattern that differs from that typically seen during experimental nociceptive pain. In this study, we used positron emission tomography to measure changes in regional cerebral blood flow (rCBF) in patients with clinical allodynia. Nine patients with peripheral nerve injury were scanned during rest, brush-evoked allodynia, and brushing of normal contralateral skin. PET data were analyzed for the whole group and for single subjects. Allodynic stimulation activated the contralateral orbitofrontal cortex (BA 11) in every patient. Whereas normal brushing activated most strongly the contralateral insular cortex, allodynic brushing produced an ipsilateral activation in this area. Another important difference between normal and allodynic brushing was the absence of a contralateral primary somatosensory cortex (SI) activation during allodynic brushing. No thalamic activation was observed during allodynic or control brushing. Although no anterior cingulate cortex (ACC) activation could be demonstrated in the group analysis, single subject analysis revealed that four patients activated this region during brush evoked allodynia. A direct post hoc comparison of brush -and allodynia-induced rCBF changes showed that allodynia was associated with significantly stronger activations in orbitofrontal cortex and ipsilateral insula whereas non-painful brushing more strongly activated SI and BA 5/7. These findings indicate that activity in the cortical network involved in the sensory-discriminative processing of nociceptive pain is downregulated in neuropathic pain. Instead, there is an upregulation of activity in the orbitofrontal and insular cortices, which is probably due to the stronger emotional load of neuropathic pain and higher computational demands of processing a mixed sensation of brush and pain. PMID- 16368193 TI - Activity of bleach, ethanol and two commercial disinfectants against spores of Encephalitozoon cuniculi. AB - Encephalitozoon cuniculi is a small protist parasite in the phylum Microspora. Hosts are infected by ingestion or inhalation of spores passed in the urine or feces. Infection with E. cuniculi is usually asymptomatic, except in young or immunocompromised hosts. This study examined the effects of various disinfectants on in vitro infectivity of E. cuniculi spores. Spores of E. cuniculi were exposed to several dilutions of commercial bleach, 70% ethanol and dilutions of commercial disinfectants HiTor and Roccal for 10 min and then loaded onto human fibroblast cells (Hs68 cells). Ten minutes of exposure to these disinfectants was lethal to E. cuniculi spores. Additional exposure time studies were done using dilutions of bleach at 0.1, 1 and 10%, and 70% ethanol. Exposure of E. cuniculi spores to 1 or 10% bleach for 30s rendered them non-infectious for Hs68 cells. Growth of E. cuniculi was observed in Hs68 cells inoculated with spores treated with 0.1% bleach for 30s or 1, 3 and 5 min, but not with spores treated for 7 min or longer. Exposure of E. cuniculi spores to 70% ethanol for 30s rendered them non-infectious for Hs68 cells. Spores of E. cuniculi are more sensitive to disinfectants than are coccidial oocysts and other parasite cysts. The relatively short contact time needed to kill spores indicates that disinfection of animal housing may be a viable means to reduce exposure of animals to E. cuniculi spores. PMID- 16368194 TI - Cultured human epithelium: human umbilical cord blood stem cells differentiate into keratinocytes under in vitro conditions. AB - BACKGROUND: Stem cells have the capacity to renew or to give rise to a specialized cell types. Human umbilical cord blood (HUCB) has been explored as an alternative source of stem cells. However, its potential to differentiate into cells of other tissues is still under discussion. The aim of our study was to evaluate if HUCB stem cells could differentiate into epithelial cells under in vitro conditions. METHODS: Human keratinocytes derived from adult female skin donors, were isolated and cultured on fibrin glue/fibroblast gels-control group. In the umbilical cord blood cell group, male umbilical cord blood cells were added at a 1:10 ratio to keratinocytes and co-cultured on the fibrin glue/fibroblasts gel. After 15 days of culture, the sheets were analyzed by use of histochemistry and FISH. DNA was extracted and evaluated by use of polymerase chain reaction (PCR) for detection of Y-chromosome-specific sequences. RESULTS: In both groups a regular epithelial sheet consisting of three to four layers of cells was formed. Using PCR and FISH, in the umbilical cord blood cell group the presence of Y-chromosome-specific sequences in the cultured keratinocytes could be detected. In the control group, no Y-chromosome-specific sequences could be detected. CONCLUSION: Our findings indicate that umbilical cord blood stem cells differentiate into epithelial cells under in vitro conditions and thereby, might serve as a starting material for isolation and expansion of cells for transplantation in patients with large skin defects. PMID- 16368195 TI - Distribution of cannabinoid receptor 1 in the CNS of zebrafish. AB - The cannabinoid receptor 1 (Cb1) mediates the psychoactive effect of marijuana. In mammals, there is abundant evidence advocating the importance of cannabinoid signaling; activation of Cb1 exerts diverse functions, chiefly by its ability to modulate neurotransmission. Thus, much attention has been devoted to understand its role in health and disease and to evaluate its therapeutic potential. Here, we have cloned zebrafish cb1 and investigated its expression in developing and adult zebrafish brain. Sequence analysis showed that there is a high degree of conservation, especially in residues demonstrated to be critical for function in mammals. In situ hybridization revealed that zebrafish cb1 appears first in the preoptic area at 24 hours post-fertilization. Subsequently, transcripts are detected in the dorsal telencephalon, hypothalamus, pretectum and torus longitudinalis. A similar pattern of expression is recapitulated in the adult brain. While cb1 is intensively stained in the medial zone of the dorsal telencephalon, expression elsewhere is weak by comparison. In particular, localization of cb1 in the telencephalic periventricular matrix is suggestive of the involvement of Cb1 in neurogenesis, bearing strong resemblance in terms of expression and function to the proliferative mammalian hippocampal formation. In addition, a gradient-like expression of cb1 is detected in the torus longitudinalis, a teleost specific neural tissue. In relation to dopaminergic neurons in the diencephalic posterior tuberculum (considered to be the teleostean homologue of the mammalian midbrain dopaminergic system), both cb1 and tyrosine hydroxylase-expressing cells occupy non-overlapping domains. However there is evidence that they are co-localized in the caudal zone of the hypothalamus, implying a direct modulation of dopamine release in this particular region. Collectively, our data indicate the propensity of zebrafish cb1 to participate in multiple neurological processes. PMID- 16368196 TI - Nitric oxide from the laterodorsal tegmental neurons: its possible retrograde modulation on norepinephrine release from the axon terminal of the locus coeruleus neurons. AB - Nitric oxide released from the cholinergic neurons in the pons may play important roles in sleep-wake regulation. However, there are few reports demonstrating the mechanisms of nitric oxide release in the cholinergic neurons in the pons. The present study investigated the effects of drug delivery of N-methyl-D-aspartic acid on nitric oxide and the neurotransmitters released in the laterodorsal tegmental nucleus (LDT), one of the major cholinergic cell groups in the pons, in rats by in vivo microdialysis with a view to clarifying nitric oxide functions in the cholinergic system. The application of N-methyl-D-aspartic acid (1 mM) into the LDT induced a significant increase in NO(2)and NO(3) for 40 min (P<0.001). Furthermore the same dose of N-methyl-D-aspartic acid induced a significant increase in cyclic GMP for 30 min (P<0.05), as well as in acetylcholine (P<0.001) and norepinephrine for 15 min (P<0.001). 3-(4-Morpholinyl)-sydonone imine hydrochloride (a nitric oxide donor, 5 mM) also induced significant increase in norepinephrine (P<0.05). Pretreatment with 1 mM 2-amino-5-phosphonopentanoic acid (an antagonist of N-methyl-D-aspartic acid receptor) prevented the N-methyl-D aspartic acid-induced increase in cyclic GMP (P<0.01), acetylcholine and norepinephrine (P<0.01), while that with 1 mM N(G)-nitro-L-arginine (an inhibitor of nitric oxide synthase) prevented the increase in cyclic GMP (P<0.01) and norepinephrine (P<0.01) but not in acetylcholine. These results suggested that nitric oxide release in the LDT induced by activation of the N-methyl-D-aspartic acid receptor on the cholinergic neurons of the LDT, then through the cyclic GMP system, facilitates norepinephrine release from the terminals of noradrenergic neurons in the locus coeruleus. Based on these findings, we propose a possible role of nitric oxide in the LDT is as a retrograde regulator of norepinephrine release from the locus coeruleus. PMID- 16368197 TI - Imiquimod: a potential weapon against Dupuytren contracture. AB - Dupuytren disease is a proliferative fibroplasia of the subcutaneous palmar tissue, occurring in the form of nodular and cords. Evidence is certainly accumulating for raised levels in Dupuytren's tissue of growth factors known to stimulate fibroblasts, Interleukin-1, basic fibroblast growth factor, transforming growth factor-beta, prostaglandin-F2, prostaglandin-E2, platelet derived growth factor and connective tissue growth factor have been suggested to have a role. Immune modification of profibrotic cytokines would provide a novel means to treat dupuytren contracture. Imiquimod cream 5% (Aldara) is an immune modifier, that downregulates transforming growth factor-beta and fibroblast growth factor-2 (the two most important cytokine in producing fibrosis). Based on previous mentioned evidence we suggest: imquimod as a potential drug for dupuytren contracture treatment. PMID- 16368198 TI - Science as a general education: Conceptual science should constitute the compulsory core of multi-disciplinary undergraduate degrees. AB - It is plausible to assume that in the future science will form the compulsory core element both of school curricula and multi-disciplinary undergraduate degrees. But for this to happen entails a shift in the emphasis and methods of science teaching, away from the traditional concern with educating specialists and professionals. Traditional science teaching was essentially vocational, designed to provide precise and comprehensive scientific knowledge for practical application. By contrast, future science teaching will be a general education, hence primarily conceptual. Its aim should be to provide an education in flexible rationality. Vocational science teaching was focused on a single-discipline undergraduate degree, but a general education in abstract systematic thinking is best inculcated by studying several scientific disciplines. In this sense, 'science' is understood as mathematics and the natural sciences, but also the abstract and systematic aspects of disciplines such as economics, linguistics, music theory, history, sociology, political science and management science. Such a wide variety of science options in a multi-disciplinary degree will increase the possibility of student motivation and aptitude. Specialist vocational science education will progressively be shifted to post-graduate level, in Masters and Doctoral programs. A multi-disciplinary and conceptually-based science core curriculum should provide an appropriate preparation for dealing with the demands of modern societies; their complex and rapidly changing social systems; and the need for individual social and professional mobility. Training in rational conceptual thinking also has potential benefits to human health and happiness, since it allows people to over-ride inappropriate instincts, integrate conflicting desires and pursue long-term goals. PMID- 16368199 TI - Combining psychometric and biometric measures of substance use. AB - This paper examines the need, feasibility, and validity of combining two biometric (urine and saliva) and three self-report (recency, peak quantity, and frequency) measures of substance use for marijuana, cocaine, opioids, and other substances (including alcohol and other drugs). Using data from 337 adults with substance dependence, we used structural equation modeling to demonstrate that these multiple measures are driven by the same underlying factor (substance use) and that no single measure is without error. We then compared the individual measures and several possible combinations of them (including one based on the latent factors and another based on the Global Appraisal of Individual Needs (GAIN) Substance Frequency Scale) to examine how well each predicted a wide range of substance-related problems. The measure with the highest construct validity in these analyses varied by drug and problem. Despite their advantages for detection, biometric measures were frequently less sensitive to the severity of other problems. Composite measures based on the substance-specific latent factors performed better than simple combinations of the biometric and psychometric measures. The Substance Frequency Scale from the GAIN performed as well as or better than all measures across problem areas, including the latent factor for any use. While the research was limited in some ways, it has important implications for the ongoing debate about the proper way to combine biometric and psychometric data. PMID- 16368200 TI - Cloning, characterization and tissue expression of rat SULT2B1a and SULT2B1b steroid/sterol sulfotransferase isoforms: divergence of the rat SULT2B1 gene structure from orthologous human and mouse genes. AB - cDNAs for rat SULT2B1 steroid/sterol sulfotransferase isoforms were cloned, and the encoded proteins overexpressed, purified and characterized. The rat SULT2B1a isoform avidly sulfonates pregnenolone but poorly utilizes cholesterol as a substrate, whereas cholesterol is more efficiently sulfonated than pregnenolone by the SULT2B1b isoform; on the other hand, neither isoform sulfonates dehydroepiandrosterone to any significant degree. Real-time PCR revealed that SULT2B1a was only expressed in brain and testis, whereas SULT2B1b was mainly expressed in skin, intestine and kidney. The SULT2B1 gene is unique among steroid/sterol sulfotransferase genes in that it encodes for two isoforms as a result of an alternative exon I. Interestingly, whereas the orthologous human and mouse SULT2B1 gene structures are identical, the rat SULT2B1 gene structure diverges. Similar to human and mouse SULT2B1 genes the rat SULT2B1 gene consists of an alternative exon I; however, as a result of exonic rearrangement, the genic locations of exons IA and IB are reversed in the rat gene. Where exon IA is located downstream of exon IB in the human and mouse SULT2B1 genes, in the rat SULT2B1 gene exon IA is located upstream of exon IB. Furthermore, unlike the case with human and mouse SULT2B1 genes where differential splicing is necessitated since a portion of exon IA is fused with exon IB to complete the SULT2B1b mRNA, this step is not required with the rat gene. PMID- 16368201 TI - Activation of transcriptional activities of AP1 and SRE by a novel zinc finger protein ZNF445. AB - Zinc finger proteins play important roles in various cellular functions, including cell proliferation, differentiation, and apoptosis. Mitogen-activated protein kinase (MAPK) signal transduction pathways are one of the most common mechanisms in eukaryotic cell regulation. Many transcription factors are important targets of MAPKs. In this study, we identified a novel gene encoding a zinc finger protein named ZNF445. The ZNF445 mRNA consists of 9105 nucleotides and has a 1031-amino acid open reading frame. The predicted 119-kDa protein contains a leucine-rich region (LER or SCAN domain) at the N-terminus, followed by a well-conserved Kruppel-associated box (KRAB) domain. At the C-terminus of the protein, there are 14 C2H2 (Cys2-His2) zinc finger motifs. ZNF445 gene is mapped to chromosome 3p21.32. Northern blot analysis indicates that a 9.1 kb transcript specific for ZNF445 is expressed in uterus, thymus, small intestine, colon, pancreas, peripheral blood leukocyte, and especially at a higher level in the testis and skeletal muscle in human adult tissues. ZNF445 protein was located in the nucleus when overexpressed in cultured cells. Reporter gene assays showed that ZNF445 is a transcriptional repressor, and overexpression of ZNF445 in the HEK 293T cells activates the transcriptional activities of AP1 and SRE. Deletion studies showed that the SCAN domain of ZNF445 may be involved in this activation. Furthermore, we found that expression of ZNF445 can increase p42/44 MAPK, MEK and Raf-1 phosphorylation. These results clearly indicate that ZNF445 is a member of the zinc finger transcription factor family and may function in MAPK pathway through Raf-1/MEK/p42/44 MAPK signals. PMID- 16368202 TI - The rise and fall of the CR1 subfamily in the lineage leading to penguins. AB - The evolution of penguins has been investigated extensively, although inconclusively, by morphologists, biogeographers and molecular phylogeneticists. We investigated this issue using retroposon analysis of insertions of CR1, which is a member of the LINE (long interspersed element) family, in the genomes of penguins and penguin relatives. The retroposon method is a powerful tool for identifying monophyletic groups. Because retroposons often show different relative frequencies of retroposition during evolution, it is first necessary to identify a certain subgroup that was specifically active during the period when the species in question diverged. Hence, we systematically analyzed many CR1 members isolated from penguin and penguin-related genomes. These CR1s are divided into at least three distinct subgroups that share diagnostic nucleotide insertions and/or deletions, namely, penguin CR1 Sph I, Sph II type A and Sph II type B. The analysis of the inserted retroposons by PCR revealed that different CR1 subfamilies or types had amplified at different rates among different periods during penguin evolution. Namely, the penguin CR1 Sph I subfamily had higher rates of retroposition in a common ancestor of all orders examined in this study or at least in a common ancestor of all extant penguins, and the subfamily Sph II type A also had the same tendency. Therefore, these CR1 members can be used to elucidate the phylogenetic relationships of Sphenisciformes (penguins) among different avian orders. In contrast, the penguin CR1 Sph II type B subfamily had higher rates of retroposition just before and after the emergence of the extant genera in Spheniscidae, suggesting that they are useful for elucidating the intra relationships among extant penguins. This is the first report for the characterization among the members of CR1 family in avian genomes excluding those of chickens. Hence, this work will be a cornerstone for elucidating the phylogenetic relationships in penguin evolution using the retroposon method. PMID- 16368203 TI - Preparation of controlled release microspheres using supercritical fluid technology for delivery of anti-inflammatory drugs. AB - Ethylcellulose/methylcellulose blends were produced using different precipitation techniques and impregnated with naproxen, a non-steroidal anti-inflammatory drug (NSAID). Solvent-evaporation technique was used not only for the preparation of ethylcellulose/methylcellulose microspheres but also to encapsulate naproxen. Supercritical fluid (SCF) impregnation was also performed to prepare naproxen loaded microspheres. The microspheres, impregnated by the SCF technique, were prepared both by solvent-evaporation and by a supercritical antisolvent (SAS) process. In vitro release profiles at pH 7.4 and 1.2, of naproxen-loaded microspheres were evaluated and the results were modelled Fick's law of diffusion and Power law. Miscrospheres prepared by supercritical antisolvent have a higher loading capacity and present a slower release profile. The systems studied present a release mechanism controlled by drug diffusion which complies Fick's law of diffusion. PMID- 16368204 TI - Anti-HIV-1 activities of extracts from the medicinal plant Rhus chinensis. AB - Rhus chinensis, a species used in folk medicine by Chinese native people, the anti-HIV-1 activities of the petroleum ether, ethyl acetate, butanol and aqueous extract of Rhus chinensis, named as RC-1, RC-2, RC-3 and RC-4, respectively, was evaluated. The petroleum ether extract RC-1 can inhibit the syncytium formation and HIV-1 p24 antigen at non-cytotoxic concentrations, the 50% effective concentration (EC50) were 0.71 and 0.93 microg/ml, respectively. The therapeutic index (TI) was about 100. RC-1 had no activity on inhibiting HIV-1 recombinant RT and HIV-1 entry into host cells. Results showed that RC-1 was effective against HIV-1 and Rhus chinensis would be a useful medicinal plant for the chemotherapy of HIV-1 infection. RC-1 might inhibit the post steps or target the new sites of HIV-1 replication. PMID- 16368205 TI - Antiplasmodial activity and cytotoxicity of plants used in West African traditional medicine for the treatment of malaria. AB - Eighteen plants originating from Ivory Coast were selected by ethnobotanical survey as plants commonly used by traditional healers for the treatment of malaria. Extracts of these plants were tested on two strains of Plasmodium falciparum: FcM29-Cameroon (chloroquine-resistant strain) and a Nigerian chloroquine-sensitive strain. The powdered plants were used to prepare three kinds of extracts: by decoction in water, in ethanol (95%) and in pentane. A radioactive micromethod allowed the evaluation of the antiplasmodial in vitro activity of the extracts on P. falciparum. Concentrations inhibiting 50% of the parasite growth (IC50) ranged from 18 microg/ml to more than 500 microg/ml for aqueous and ethanol extracts and from 4.3 microg/ml to more than 500 microg/ml for pentane extracts. Cytotoxicity was estimated on A375 melanoma cells and a cytotoxicity/antiplasmodial index (CAR) was calculated for each extract, ranging from 1 to 10. The pentane extracts of Cola caricaefolia and Uvaria afzelii, which revealed the strongest antiplasmodial activity had CAR values of about 10. PMID- 16368206 TI - Tracheo-arterial fistula in tracheostomy patients with Duchenne muscular dystrophy. AB - A tracheo-arterial fistula is a serious and life threatening potential complication of a tracheostomy. Since 1984, we experienced nine fatal cases of tracheo-arterial fistula among 60 Duchenne muscular dystrophy (DMD) patients who underwent a tracheostomy. Representative cases included a patient with lordosis (Case 8), in whom the fistula was located in the brachiocephalic artery close to the trachea, and another with severe scoliosis (Case 9), which caused the aorta to compress the trachea. Such anatomical changes can be the cause of a fistula between the trachea and brachiocephalic artery. The anatomical locations between the trachea and brachiocephalic artery are modified by thoracic deformities in DMD patients, and should be confirmed using computed tomography (CT) prior to a tracheostomy procedure. Further, during such a procedure, the tracheal stoma must be placed in a location clearly away from the arteries, and should be followed by regular post-operative examinations using CT and careful management to avoid a tracheo-arterial fistula. PMID- 16368207 TI - Global developmental delay, osteopenia and ectodermal defect: a new syndrome. AB - Global developmental delay is a serious social problem. It is often unrecognized and the phenotypes are inadequately studied. To investigate the phenotypes of children with aspecific central nervous system (CNS) impairment (poor speech, maladaptive behavioral symptoms such as temper tantrums, aggressiveness, poor concentration and attention, impulsiveness, and mental retardation). SETTING: Tertiary care hospital. PATIENTS: Three children (two male siblings, and one unrelated girl). METHODS: We used the results from clinical neurological evaluations; imaging and electrodiagnostic studies; metabolic and genetic tests; skin biopsies and bone mineral densitometry. All three children suffered from (A) global developmental delay, (B) osteopenia, and (C) identical skin defects. The skin ultrastructural abnormalities were abnormal keratin differentiation, consisting of hyperkeratosis and granular layer thickening; sweat gland abnormalities, consisting of focal, cytoplasmic clear changes in eccrine secretory cells; and melanocyte abnormalities, with both morphological changes (reduced number and size without evident dendritic processes), and functional changes (defects in the migration of melanosomes in the keratinocytes). These patients present a previously unrecognized syndrome. We retain useful to report this new association, to be recognized, in the next future, as a specific key sign of a well-defined genetic defect. PMID- 16368208 TI - Cerebral oxygen supply during hypotension in near-term lambs: a near-infrared spectroscopy study. AB - Sufficient O(2)-supply to the brain is necessary for an adequate cerebral energy metabolism, function and growth. To elucidate the relation between changes in, respectively, mean arterial blood pressure (MABP) and cerebral O(2)-supply and changes in the oxygenation state of hemoglobin during hypotension in preterm born lambs. Preterm lambs were delivered at 141 days (n=7) or 127 days (n=7) of gestation. Hypotension was induced by stepwise withdrawal of blood. Cerebral arterial blood gases were analyzed at the end of each level to calculate cerebral O(2)-supply. Near-infrared spectroscopy was used to measure changes in the concentration of cerebral oxyhemoglobin (cO(2)Hb), deoxyhemoglobin (cHHb) and cHbD (the difference between cO(2)Hb and cHHb). In the 141 and the 127 d lambs, changes in MABP and cerebral O(2)-supply were positively linearly related with DeltacO(2)Hb, and negatively with DeltacHHb. MABP was positively linearly related with changes in cHbD. During hemorrhagic hypotension, changes in MABP and cerebral O(2)-supply are reflected by changes in the oxygenation state of cerebral hemoglobin in near-term born lambs. PMID- 16368209 TI - Efficacy and adverse effects of rectal thiamylal with oral triclofos for children undergoing magnetic resonance imaging. AB - We studied the efficacy and adverse effects of rectal thiamylal in combination with oral triclofos in sedation for pediatric magnetic resonance imaging. Five hundred forty-six children underwent MRI examination from January of 1997 to December of 2001. Among them, 10mg/kg of rectal thiamylal was administrated after oral triclofos in 378 children. Successful sedation was obtained in 321 of 378 patients (85%) after a single rectal administration of thiamylal. Totally, 369 children (98%) could undergo MRI examination completely under successful sedation. Adverse effect was observed only in one patient showing respiratory depression. Rectal thiamylal is effective for sedation for MRI in children. Adverse effect was rare in our patients. Although the risk of side effect was considered to be rare, we should follow principles for the sedation of children. PMID- 16368210 TI - Pseudotumor cerebri as an important differential diagnosis of papilledema in children. AB - INTRODUCTION: Primary pseudotumor cerebri (PTC) in childhood is a rare but important differential diagnosis in children presenting with papilledema. It is defined as elevated cerebrospinal fluid (CSF) pressure of more than 20 cm H(2)O, normal CSF composition, and exclusion of underlying structural or systemic causes. Visual loss is a serious complication, which requires careful monitoring and management. PATIENTS AND METHODS: We conducted a retrospective chart review of 12 patients with primary PTC. The mean age at presentation was 8212 years, and there was a male-to-female ratio of 7:5. The aim of this study was to investigate the clinical features of primary PTC in children, and to highlight the different treatment options in normalizing intracranial pressure in these patients. RESULTS: In the majority of cases, children presented with headache. Four patients had no obvious symptoms and papilledema was found on routine eye examination. Obesity was uncommon and there was no distinct sex predilection. Acetazolamide was our drug of choice for the initial treatment. Furosemide and prednisone were used as second-line agents. Treatment was gradually decreased after resolution of the papilledema with exception of the two youngest children, who remained symptomatic. One child underwent ventricular-peritoneal shunting. DISCUSSION: The treatment goals of PTC are the relief of symptoms, and preservation of visual function. Acetazolamide is an effective first-line method of lowering raised intracranial pressure. In our study group especially the young children were difficult to treat. This might indicate an age-related difference in the etiology of PTC. When medical treatment remains ineffective and visual function deteriorates, surgical treatment should be considered. PMID- 16368211 TI - Maternal swimming during pregnancy enhances short-term memory and neurogenesis in the hippocampus of rat pups. AB - In the present study, the effects of maternal swimming during pregnancy on the short-term memory ability, hippocampal neurogenesis, and brain-derived neurotrophic factor (BDNF) mRNA expression of rat pups were investigated. After confirming their pregnancy, the pregnant rats were divided into two groups: the control group and the swimming group. From the 15th day of pregnancy until delivery, pregnant rats were subcutaneously injected with 100mg/kg of 5-bromo-2' deoxyuridine (BrdU) once a day at 30min before the starting of swimming exercise. Pregnant rats in the swimming group were forced to swim for 10min once a day until delivery. On the 21 days after birth, the rat pups were trained in a step down avoidance test. The latency time of the step-down avoidance task was determined on the 28 days after birth in order to evaluate the short-term memory ability of pups. On the 29 days after birth, the rat pups' brains were removed, and BrdU immunohistochemistry for the detection of neurogenesis and reverse transcription-polymerase chain reaction (RT-PCR) for the detection of BDNF mRNA expression were then performed. The rat pups born from the maternal rats that performed swimming during pregnancy showed significantly increased BDNF mRNA expression, enhanced hippocampal neurogenesis, and improved short-term memory capability. The present results have clearly shown that maternal swimming by rats during pregnancy enhances the memory of the rats' offspring by increasing neurogenesis. Our present study provides the evidence that maternal exercise during the gestational period may enhance the brain functions of the mothers' offspring. PMID- 16368212 TI - An adolescent with pharyngeal-cervical-brachial variant of Guillain-Barre syndrome after cytomegalovirus infection. AB - A 15-year-old Japanese girl developed bulbar palsy and upper limb-dominant muscle weakness 2 weeks after the onset of an upper respiratory tract infection due to cytomegalovirus (CMV). Her symptoms resembled that seen in the pharyngeal cervical-brachial variant (PCB) of Guillain-Barre syndrome (GBS). Although bulbar palsy usually continues for several months in PCB, her bulbar palsy was very mild and improved rapidly before intravenous immunoglobulin therapy was instituted. Serum anti-GT1a IgG antibody titer was elevated at the acute phase of the disease and gradually decreased. The bulbar palsy-dominant GBS is thought to relate to anti-GT1a antibody and Campylobacter jejuni infection in adult patients. Our Case report suggests that CMV can also induce the production of anti-GT1a antibody, thereby resulting in PCB. When one sees acute onset bulbar palsy and limb muscle weakness, the possibility of PCB, even in children, should be considered, thus compelling the need for serum anti-ganglioside antibody measurement. PMID- 16368213 TI - Arthrogryposis multiplex congenita with callosal agenesis and dentato-olivary dysplasia. AB - We report the autopsy case of a boy with arthrogryposis multiplex congenita, associated with callosal agenesis and dentato-olivary dysplasia. The patient manifested with dysmorphic facial features and suffered from intractable epilepsy during the neonatal period. These sets of complications suggest that a common molecular mechanism may be involved in the development of corpus callosum and the folding of the dentate and inferior olivary nuclei. Deep brain structures, including the brainstem and the cerebellum, may be involved in the pathophysiology of symptomatic generalized epilepsy. The differential diagnoses for the clinical and pathological characteristics of this patient are discussed. PMID- 16368214 TI - Forced mouth opening reaction: a primitive reflex released from cortical inhibition. AB - We report the case of a 6-year-old girl with congenital adrenal hyperplasia, who showed 'forced mouth opening reaction' during the course of acute encephalopathy due to adrenal crisis. When an object was moved towards her mouth, or when the corner of her mouth was stroked with a tongue depressor, she would immediately open her mouth fully and hold it open. This reaction appeared transiently during the course of her illness in association with other frontal release signs including the rooting, groping and palmomental reflexes. Magnetic resonance imaging showed bilateral widespread lesions involving the gray and white matters in the frontal lobes, and less severe lesions in the temporal and parietal areas. We propose that this unique reaction is a sign of a release phenomenon, and represents the emergence of primitive reflexes in the absence of cortical inhibition in some types of encephalopathies. PMID- 16368215 TI - Subacute sclerosing panencephalitis surveillance study in Istanbul. AB - The exact incidence rate of subacute sclerosing panencephalitis (SSPE) in Turkey (and in Istanbul) is not known. We have conducted an active surveillance study to determine the epidemiological characteristics and the incidence rate of SSPE in Istanbul between the dates July 1, 2002 and July 1, 2004. We found that the incidence of SSPE in Istanbul is 2 per million. By logistic regression analysis, risk factors in SSPE development are determined as being at younger ages (OR: 1.199, 95%CI=1.047-1.372, P=0.009), living in crowded households (OR: 1.430, 95%CI=1.039-1.968, P=0.028), low education level of the mother (OR: 0.123, 95%CI=0.034-0.447, P=0.001), low household income (OR: 0.413, 95%CI=0.234-0.728, P=0.002), infant's being born out of Marmara region (Istanbul is in Marmara region of Turkey) (OR: 0.358, 95%CI: 0.172-0.746, P=0.006), infant's not being vaccinated against measles (OR: 0.495, 95%CI: 0.312-0.786), infant's having had measles before (OR: 0.235, 95%CI: 0.135-0.411). As a result, it is found in this study that SSPE is mostly related to having measles infection, and measles vaccination is found to be highly protective against SSPE. This is the first epidemiological study in SSPE from Turkey that conveys the incidence rate in Istanbul. PMID- 16368216 TI - Glutaric aciduria types I and II. AB - Glutaric aciduria type I is an autosomal recessive disorder resulting from a deficiency of glutaryl-CoA dehydrogenase. This leads to an accumulation of glutaric and 3-hydroxyglutaric acids and secondary carnitine deficiency. The symptomatology is discussed, especially those resulting from lesions in the basal ganglia, and the encephalopathic episodes which are often precipitated by infections. The variability of the clinical presentation is stressed. The most serious complications are collections of fluid and blood in the middle fossae, the bleeding resulting from rupture of bridging veins. The prognosis does not seem to be related to the extent of the enzyme deficiency. The diagnosis is confirmed by identifying the abnormal acids in the urine and the deficiency of the enzyme in cultured fibroblasts. The differential diagnosis is reviewed: from other biochemical disorders and from other cerebral lesions. Treatment is by special diet and carnitine supplementation. The dystonia can prove difficult to treat, and surgery may be needed to remove the collections of fluid and blood. Glutaric aciduria type II is caused by a deficiency of either electron transport flavoprotein or of electron transport flavoprotein oxoreductase. The symptoms can be mild or severe. The former may only occur in times of stress, and the latter include congenital anomalies, especially of the kidneys and heart. The pathology of these are discussed. The demonstration of organic acids in the urine and the results of muscle and liver biopsies confirm the diagnosis, and treatment with a special diet and supplementation with carnitine and riboflavine is effective. PMID- 16368217 TI - Brain MRI abnormalities in muscular dystrophy due to FKRP mutations. AB - INTRODUCTION: FKRP mutations cause a muscular dystrophy which may present in the neonatal period (MDC1C) or later in life (LGMD2I). Intelligence and brain imaging have been previously reported as being normal in FKRP-associated muscular dystrophy, except in rare cases presenting with mental retardation associated with structural brain abnormalities. PATIENTS AND METHODS: We studied cerebral MRIs in twelve patients with FKRP-associated muscular dystrophy presenting in infancy or early childhood, at ages between 14 months and 43 years. Two patients had severe cognitive deficits, four had mild-moderate mental retardation and the rest were considered to have normal intelligence. All, but one were wheelchair bound and 7 were mechanically ventilated. RESULTS: Brain MRI was abnormal in 9 of 12 patients. Brain atrophy was seen in 8 patients. One child had isolated ventricular enlargement at 4 years. Cortical atrophy involved predominantly temporal and frontal lobes and was most important at later ages. In two cases with serial images this atrophy seemed progressive. Three patients, two with severe and one with moderate mental retardation, showed structural abnormalities of the posterior fossa with hypoplasia of the vermis and pons, and cerebellar hemispheric cysts. These abnormalities were stable with time. Two of these three patients also showed diffuse white matter abnormalities in early childhood, which regressed with time. CONCLUSIONS: MRI abnormalities are common in patients with FKRP-associated muscular dystrophy presenting at birth or in early childhood. Progressive brain atrophy is the most frequent finding. Posterior fossa malformations and transient white matter changes may be seen in patients with associated mental retardation. PMID- 16368218 TI - Incidence of chronic knee lesions in long-distance runners based on training level: findings at MRI. AB - PURPOSE: The purpose of this study was to evaluate the incidence of chronic knee changes in long-distance runners based on the training status, including distance, running frequency, training pace, and running experience. METHODS: MRI of the knee was performed in 26 non-professional runners 5 days after their last training unit. Lesions of the menisci and cartilage (5-point scale), bone marrow and ligaments (3-point scale), and joint effusion were evaluated. A total score comprising all knee lesions in each runner was evaluated. The incidence of the knee changes was correlated with the training level, gender, and age of the runners. RESULTS: Grade 1 lesions of the menisci were found in six runners with a high training level, and in only four runners with a low training level. Grade 1 cartilage lesions were found in three high-trained runners and in one low-trained runner, and grade 2 lesions were found in one high-trained runner and in two low trained runners, respectively. Grade 1 anterior cruciate ligament lesions were seen in three runners with a high- and in two runners with a low-training level. Runners with a higher training level showed a statistically significant higher score for all chronic knee lesions than those with a lower training level (p<0.05). CONCLUSIONS: MRI findings indicate that a higher training level in long distance runners is a risk factor for chronic knee lesions. PMID- 16368219 TI - HPLC determination of phenolic acids and antioxidant activity in concentrated peat extract--a natural immunomodulator. AB - The aim of this study was to devise a method for identification and quantification of phenolic acids in concentrated peat extract samples. The simple reversed-phase HPLC method for simultaneous determination of several phenolic acids was developed. The method was validated and it was suitable for the analysis of phenolic acids in peat extracts. This method allowed identifying eight phenolic acids in peat extracts. Phenolic profiles of two samples of peat extract obtained from different medicinal peats were similar, although variations in amounts of individual phenolic acids were observed. Also, slight variations in total phenolic content were detected. The antioxidant activity of peat extracts was evaluated with spectrophotometric ABTS assay. Differences in antioxidant activity were observed for two samples of peat extract produced from different peat varieties. This differences probably reflected phenolic composition of peat extracts. PMID- 16368220 TI - Optimal matches of patient preferences for information, decision-making and interpersonal behavior: evidence, models and interventions. AB - OBJECTIVE: A comprehensive review was conducted of the theoretical and empirical work that addresses the preference-match strategy in physician-patient communication. METHODS: Searches were conducted on Medline, PsychINFO, InFoTrac One File Plus, Sociological Abstracts, and Dissertation Abstracts through 2004. The following keywords were used: patient preferred and received information; patient preferred and actualized treatment decision-making; patient-physician beliefs in shared decision-making; patient-physician match, fit, or concordance; reciprocal relationship or mutuality; doctor-patient affiliation, control, relationship; match/fit between patient and physician in affiliation, control, or relationship. RESULTS: Findings revealed varying degrees of support for the positive effects of matching patients' preferred levels of information, decisional control, and consultative interpersonal behavior. CONCLUSIONS: Findings justify not only continued but expanded research efforts in this area that would incorporate recommended changes in research design and implementation. PRACTICE AND RESEARCH IMPLICATIONS: Assessment strategies and match interventions are discussed that, if evidence continues to be supportive, might routinely optimize patient-physician encounters toward more positive outcomes. Methodological guidelines are suggested that can improve future preference-match studies of the patient-physician interaction. Practitioners need to consider adoption of patient-match assessment and intervention strategies in addition to recent exclusive concentrations on patient-centered and shared decision-making approaches. PMID- 16368221 TI - The psychometric properties of the Brief Fear of Negative Evaluation Scale. AB - This study examined the psychometric properties of the Brief Fear of Negative Evaluation (BFNE) Scale in a nonclinical, nonstudent sample. Participants were administered the BFNE Scale, Beck Depression Inventory (BDI), and the UCLA Loneliness Scale. No differences were found across age, but women scored significantly higher on the BFNE Scale than men. Confirmatory factor analysis supported a two-factor solution corresponding to positive and reverse scored items. Internal consistency was excellent for the positive scored factor (alpha=.94), acceptable for the reverse scored factor (alpha=.73), and good for the full BFNE Scale (alpha=.80). BFNE scores were significantly correlated in the expected directions with the BDI and the UCLA Loneliness Scale. Implications of these findings for clinical practice and future research are discussed. PMID- 16368223 TI - Production of a range image for facial motion analysis: a method for analyzing lip motion. AB - To analyze the lip movement of patients with cleft lip and palate in detail, we developed a novel technique to produce a range image from a video-based motion capture system, and a unique technique to analyze lip movements in detail three dimensionally. The accuracy of the range image obtained from the present system was investigated using a positioning actuator with a high accuracy and a known object, and by comparisons with the range images produced from a laser scanner with a cast model of a face. The lip motions of six healthy adults during phonation was analyzed. PMID- 16368222 TI - Characterization of structure and expression of the Dzip1 gene in the rat and mouse. AB - A transcript encoding a rat homologue of DZIP1 (DAZ-interacting protein) was isolated from testis RNA. Like human DZIP1, it contains a C(2)H(2) zinc finger domain. A predicted mouse homologue of DZIP1 was found in the GenBank database. Genome analysis indicated that while DZIP1 and mouse Dzip1 contain 22 and 20 exons, respectively, the rat sequence was intronless, confirmed by PCR on genomic DNA. This rat Dzip1 sequence is homologous to mouse Dzip1 exons 1-6 and DZIP1 exons 5-9. As this rat sequence was shorter than DZIP1 it was designated rat Dzip1S. The rat genome also contained a further predicted homologue of DZIP1 displaying conserved linkage homology with mouse Dzip1 and DZIP1. This sequence, if expressed, is the true rat homologue of DZIP1, designated rat Dzip1. Rat Dzip1S mRNA was present in all tissues examined by qualitative RT-RCR, and in situ hybridization of rat testis confirmed that expression of rat Dzip1S mRNA was confined to the spermatogenic lineage, specifically premeiotic spermatogonia. PMID- 16368224 TI - Cell-penetrating anti-native DNA antibodies trigger apoptosis through both the neglect and programmed pathways. AB - It has been previously shown that certain antibodies to double stranded-DNA are capable to transgress the cell membrane, translocate to the cell nucleus and react with their antigens. When so doing, they signal the penetrated cells to undergo apoptosis. In this study we dissected the two main-streams of apoptotic triggering and found that the neglect, mitochondrial pathway, is definitely involved, but activation of caspases 2 and 8, classically known to be associated to the membrane pathway, also occurs in a small proportion of cells. Our results suggest that penetrating anti-dsDNA antibodies, besides activating neglect apoptosis, may also trigger programmed cell death, perhaps initiated at the cell membrane level. PMID- 16368225 TI - Cell type-specific changes in spontaneous and minimally evoked excitatory synaptic activity in hippocampal CA1 interneurons of kainate-treated rats. AB - The epileptiform activity in the kainic acid (KA) model of epilepsy arises from complex changes in excitation and inhibition. To assess the involvement of excitatory drive onto inhibitory interneurons in this epileptiform activity, we examined changes in spontaneous and minimally evoked excitatory post-synaptic currents (sEPSCs and eEPSCs) in CA1 interneurons in stratum oriens/alveus (O/A) and stratum radiatum (RAD) in rat hippocampal slices after KA treatment. The frequency and amplitude of sEPSCs and the amplitude of eEPSCs were unchanged in O/A interneurons, but the EPSC kinetics were significantly slower. These changes appear to be due to altered kinetics and voltage-dependent properties of the NMDA component of EPSCs in O/A interneurons. In contrast, sEPSCs and eEPSCs in RAD interneurons did not change after KA treatment. The distinct changes in excitatory synaptic activity in interneurons differentially involved in feedback (O/A) versus feedforward (RAD) inhibition suggest a cell type-specific reorganization of excitatory synapses after KA treatment. These modifications in excitatory input to interneurons could contribute to the maintenance of inhibition of CA1 pyramidal cells after KA treatment, or may also create network conditions favourable to epileptiform activity. PMID- 16368226 TI - Association of rpoB mutations with rifampicin resistance in Mycobacterium avium. AB - The susceptibility of clinical isolates of Mycobacterium avium to rifampicin (RIF) was examined. All 32 clinical isolates tested, including 18 from Japan, 13 from Poland and 1 from Thailand, were resistant to RIF (minimum inhibitory concentrations (MICs) > or =32 microg/mL for 17 isolates and 2-16 microg/mL for 15 isolates), whereas the type strain of M. avium ATCC 25291 was susceptible to RIF (MIC < or = 0.03 microg/mL). Mutations in nucleotides 1276-1356 of the rpoB gene, termed the 81 bp core region, are associated with RIF resistance in Mycobacterium tuberculosis. No mutations were found in this region in any of the M. avium clinical isolates tested. However, mutation of G-->A to give a Gly544- >Asp substitution was identified within the rpoB gene downstream of the 81 bp region in all clinical isolates. A RIF-resistant strain (ATCC 25291 Rif(r); MIC> or =32 microg/mL) obtained by culturing the type strain in RIF-containing broth possessed a mutation C-->T to give a His445-->Tyr substitution within the 81 bp region. When the rpoB gene of the ATCC 25291 Rif(r) strain and of a clinical isolate were inserted into Mycobacterium smegmatis, organisms with the ATCC 25291 Rif(r) sequence, but not those with the clinical isolate sequence, showed resistance to RIF. These results suggest that mutations of the 81 bp region of rpoB, as well as factors other than rpoB mutation, confer RIF resistance in M. avium. PMID- 16368227 TI - Visualizing common deletion of mitochondrial DNA-augmented mitochondrial reactive oxygen species generation and apoptosis upon oxidative stress. AB - Common deletion (CD) 4977 bp of mitochondrial DNA (mtDNA) disrupt specifically mitochondrial complex I, IV and V on the electron transport chain (ETC) and is closely associated with wide spectrums of clinical manifestations. To quantitatively investigate how CD-induced ETC defect alters mitochondrial reactive oxygen species (mROS) generation as well as down stream apoptotic signaling, we employed an established array of human CD cytoplasmic hybrids (cybrids) harboring 0%-80% of CD. Pathological effects of CD on the mitochondria were visualized at single cell level by the application of fluorescent probes coupled with conventional and multiphoton imaging microscopy. Intriguingly, we observed CD-augmented mROS generation omitted "threshold effect". CD-augmented mROS generation was associated with depolarized mitochondrial membrane potential (DeltaPsi(m)). Upon oxidative stress, the amount of CD-augmented mROS generation was greatly enhanced to cause pathological apoptotic deterioration including opening of the mitochondrial permeability transition, cytochrome c release, phosphatidylserine externalization and DNA fragmentation. In addition, heterogeneous mitochondrial dysfunctions were found in cybrids containing 80% of CD (D cybrids), i.e., low sensitive-D (LS-D, roughly 80%) and a super sensitive-D (SS-D, 20%). As compared to LS-D, SS-D had higher resting mROS level but slightly hyperpolarized DeltaPsi(m). Upon H2O2 treatment, much faster generation of mROS was observed which induced a faster depolarization of DeltaPsi(m) and later apoptotic deterioration in SS-D. We proposed a dose-dependent, feed-forward and self-accelerating vicious cycle of mROS production might be initiated in CD induced ETC defect without threshold effect. As CD-augmented mROS generation is obligated to cause an enhanced pathological apoptosis, precise detection of CD augmented mROS generation and their degree of heterogeneity in single cells may serve as sensitive pathological indexes for early diagnosis, prognosis and treatment of CD-associated diseases. PMID- 16368228 TI - Design of poly(ethylene glycol)-tethered copolymers as novel mucoadhesive drug delivery systems. AB - We developed novel acrylic-based polymers that can be used as mucoadhesive delivery systems. Poly(acrylic acid) hydrogels were modified by grafting adhesion promoter chains such as poly(ethylene glycol) (PEG) onto their back-bone chains, thus promoting the adhesive process by interpenetration. The copolymers synthesized were designated as P(AA-g-EG). Hydrogels were synthesized using PEG of two different molecular weights, 1000 and 2000, and with varying molar feed ratio of AA-EG (20:80, 40:60, 60:40, 80:20, 12:88, 25:75, 44:56, 67:33). The copolymers were synthesized by using free radical solution UV-polymerization. The effects of different PEG-tethered structures on mucoadhesion were studied using a tensiometric testing and the work of adhesion was calculated. Preswollen P(AA-g EG) copolymer films composed of 40% acrylic acid (AA) and 60% ethylene glycol (EG), containing PEG 1000 tethers, exhibited the highest value for the work of mucoadhesion, 130 x 10(-3)+/-27 x 10(-3) mJ, that is five times higher than the formulation composed of pure PAA. Based on these results and associated molecular analysis, we conclude that the higher mucoadhesive properties of this specific copolymer were the result of the synergistic effects of both monomers. AA functional groups allowed the polymer to form multiple hydrogen bonds with the glycoproteins present in the mucus. PEG tethers possibly acted as mucoadhesive promoters, enhancing interpenetration of polymer chains into the mucus. PMID- 16368229 TI - Intermediate-term toxicity of repeated orally administered doses of the anti malarial beta-artemether in dogs. AB - The artemisinin derivative beta-artemether, an anti-malarial, was evaluated for its toxicity and tolerability in a 2-week, multiple-dose study in dogs. Eight beagle dogs (4 females, 4 males) were given beta-artemether by oral gavage 3 times daily at 45 mg/kg/dosing (a total daily dose-level of 135 mg/kg) for 2 weeks. This beta-artemether dose regime was well tolerated. Body weight changes were normal although feed consumption during the treatment period reduced compared to that of the pre-trial period. Clinical signs were transient spells of soft to liquid feces. On completion of the treatment period, the animals were sacrificed and submitted to a full macroscopic post-mortem examination. Designated organs were weighed and a complete light microscopic examination was performed on 43 selected tissues from 1 animal per sex, and on the liver, kidneys, thymus, mandibular lymph nodes and lungs of the three other animals per sex. Major findings were high liver weight and histopathologic findings of slight diffuse hepatocellular hypertrophy and distal tubular dilatation, together with flattened epithelium, in the kidneys. With the dose regime used in this trial beta-artemether produced no clinical or apparent histopathological signs of neurotoxicity in dogs. PMID- 16368230 TI - The mannose-binding lectin: a prototypic pattern recognition molecule. AB - The innate immune system is comprised of a sophisticated network of recognition and effector molecules that act together to protect the host in the first minutes or hours of exposure to an infectious challenge. The mannose-binding lectin (MBL) is an evolutionary conserved circulating host defense protein that acts as a broad-spectrum recognition molecule against a wide variety of infectious agents. Target binding triggers the MBL pathway of complement activation. MBL can be considered conceptually as an 'ante-antibody' because it has a role in mammals during the lag period that is required to develop an antibody response against infectious agents. Additionally, there are MBL-like homologues in animals that lack adaptive immunity that activate a primitive complement system, and under these circumstances these MBL-like molecules play an analogous role to antibodies in higher animals. These molecules might be considered to be functional antecedents of antibodies. Recent work also indicates that MBL recognizes altered self-antigens, and as such MBL has a role that extends beyond a traditional role in first line host defense as it appears to play a role as a modulator of inflammation. PMID- 16368231 TI - Advances in preservation methods: keeping biosensor microorganisms alive and active. AB - The ability of bacteria to sense their surroundings can be employed to measure the bioavailability and toxicity of pollutants. However, long-term maintenance of both viability and activity of the sensor bacteria is required for the development of cell-based devices for environmental monitoring. To meet these demands, various techniques to conserve such bacteria have been reported, including freeze drying, vacuum drying, continuous cultivation, and immobilisation in biocompatible polymers of organic or inorganic origin. Much effort has been invested in merging these bacterial preservation schemes with the construction of sensor cell arrays on platforms such as biochips or optic fibres, hopefully leading to effective miniaturised whole-cell biosensor systems. These approaches hold much promise for the future. Nevertheless, their eventual implementation in practical devices calls for significant enhancement of current knowledge on formulation of reporter microorganisms. PMID- 16368232 TI - DNA arms do the legwork to ensure the directionality of lambda site-specific recombination. AB - The integrase protein of bacteriophage lambda (Int) catalyzes site-specific recombination between lambda phage and Escherichia coli genomes. Int is a tyrosine recombinase that binds to DNA core sites via a C-terminal catalytic domain and to a collection of arm DNA sites, distant from the site of recombination, via its N-terminal domain. The arm sites, in conjunction with accessory DNA-bending proteins, provide a means of regulating the efficiency and directionality of Int-catalyzed recombination. Recent crystal structures of lambda Int tetramers bound to synaptic and Holliday junction intermediates, together with new biochemical data, suggest a mechanism for the allosteric control of the recombination reaction through arm DNA binding interactions. PMID- 16368233 TI - Respiratory syncytial virus fusion inhibitors. Part 3: Water-soluble benzimidazol 2-one derivatives with antiviral activity in vivo. AB - The introduction of acidic and basic functionality into the side chains of respiratory syncytial virus (RSV) fusion inhibitors was examined in an effort to identify compounds suitable for evaluation in vivo in the cotton rat model of RSV infection following administration as a small particle aerosol. The acidic compounds 2r, 2u, 2v, 2w, 2z, and 2aj demonstrated potent antiviral activity in cell culture and exhibited efficacy in the cotton rat comparable to ribavirin. In a BALB/c mouse model, the oxadiazolone 2aj reduced virus titers following subcutaneous dosing, whilst the ester 2az and amide 2aab exhibited efficacy following oral administration. These results established the potential of this class of RSV fusion inhibitors to interfere with infection in vivo following topical or systemic administration. PMID- 16368234 TI - Potent 2-[(pyrimidin-4-yl)amine}-1,3-thiazole-5-carbonitrile-based inhibitors of VEGFR-2 (KDR) kinase. AB - Pyrimidino-thiazolyl carbonitriles were prepared that are potent VEGFR-2 (KDR) kinase inhibitors. The modification of lead structures resulted in 3m which exhibited the best overall profile in KDR inhibitory activity, iv/po pharmacokinetics, and reduced hERG affinity. PMID- 16368235 TI - Synthesis and antiviral activity of 2'-deoxy-2'-fluoro-2'-C-methyl purine nucleosides as inhibitors of hepatitis C virus RNA replication. AB - A series of purine nucleosides containing the 2'-deoxy-2'-fluoro-2'-C methylribofuranosyl moiety were synthesized and evaluated as potential inhibitors of the hepatitis C virus in vitro. Of the nucleosides that were synthesized, only those possessing a 2-amino group on the purine base reduced the levels of HCV RNA in a subgenomic replicon assay. PMID- 16368237 TI - Rapidly progressive neurodegeneration in a case with the 7472insC mutation and the A7472C polymorphism in the mtDNA tRNA ser(UCN) gene. AB - The authors report the clinical, neuroimaging, muscle biopsy and mtDNA findings in a patient affected by bilateral hearing loss and mental retardation since infancy, presenting at age 31 years with a rapid deterioration of mental status and ataxia leading to vegetative condition and death at the age of 32 years. Clinical and genetic studies have been also performed in the mother, affected by neurosensorial hearing loss. Muscle biopsy showed severe mitochondrial alterations in the propositus and evidence of mitochondrial alterations in his mother. Direct mtDNA sequencing in all family members revealed the known 7472insC mutation and the recently described A7472C sequence variation in the tRNA(Ser(UCN))gene. RFLP-PCR confirmed the heteroplasmic nature of the two mutations and failed to find the second transversion in 200 controls. The percentage of mutant genomes harbouring 7472insC ranged from 3 to 7% in asymptomatic family members to 70% in the proband and his mother, whereas the percentage of A7472C mutant genomes was about 90% in all maternal relatives except the proband (56%) and his sister (5%). In conclusion, this is the first report of a rapidly progressive encephalopathy in association with the 7472insC mutation in mtDNA, combined with an A>C variation at the same nucleotide with a possible suppression effect on the pathogenic mutation. PMID- 16368238 TI - Clinical or subclinical cardiac involvement in facioscapulohumeral muscular dystrophy. PMID- 16368239 TI - 138th ENMC Workshop: nemaline myopathy, 20-22 May 2005, Naarden, The Netherlands. PMID- 16368240 TI - Carbonic anhydrase inhibitors ameliorate the symptoms of hypokalaemic periodic paralysis in rats by opening the muscular Ca2+-activated-K+ channels. AB - Carbonic-anhydrase inhibitors are effective in channelopathies possibly by opening the Ca2+-activated-K+ channels. However, the in vivo effects of these drugs in K+-deficient rats, the animal model of familial hypokalaemic periodic paralysis(hypokalaemic-PP), are currently unknown. Measures of insulin-responses, serum electrolytes levels and patch-clamp experiments were therefore performed in K+ -deficient rats treated in vivo with dichlorphenamide (DCP), ethoxzolamide (ETX), hydrochlorthiazide (HCT), methazolamide (MTZ), bendroflumethiazide (BFT) and acetazolamide (ACTZ). Ten days treatments of K+-deficient rats with DCP, BFT, ETX and ACTZ (5.6 mg/kg per day) restored the serum [K+] to control values and prevented the insulin-induced paralysis. In ex vivo experiments, the carbonic anhydrase inhibitors enhanced the activity of Ca2+-activated-K+ channels with the order of efficacy: ACTZ>BFT>ETX>DCP. In contrast, HCT and MTZ failed to stimulate the Ca2+-activated-K+ channels and to prevent the hypokalaemia and paralysis. At the concentration of 1mg/kg per day, all these drugs failed to ameliorate the hypokalaemic-PP symptoms. The activation of Ca2+-activated-K+ channel in addition to the mild diuretic effect explained the efficacy of ACTZ and DCP in K+ deficient rats and in familial hypokalaemic-PP. PMID- 16368242 TI - Synthesis of 8-aminoadenosine 5'-(aminoalkyl phosphates), analogues of aminoacyl adenylates. AB - A short and efficient route for the synthesis of aminoalkyl 8-aminoadenylates, potential aminoacyl-tRNA synthetase inhibitors, is presented. Aminoalkyl 8 aminoadenylates were synthesized using a 5'-H-phosphonate strategy involving minimal protecting group manipulations and a single final deprotection step. PMID- 16368243 TI - A grid-mounted niobium body target for the production of reactive [18F]fluoride. AB - A small-volume grid-supported niobium target was constructed for the production of [18F]-fluoride on a CTI RDS 112 cyclotron. The inert properties of niobium metal have proven compatible with the superheated environment generated during proton irradiation. A grid support, constructed from aluminum, provides the necessary heat removal for high-power irradiations. The niobium and gird combination supplies reactive fluoride over an extended target lifetime. PMID- 16368244 TI - Inhibition of tumor growth in immunocompromised hosts by restoring type-2 immunity using infusion of G-CSF-treated allogeneic CD8+ leukocytes. AB - Since recombinant human granulocyte colony-stimulating factor (rhG-CSF)-treated donor leukocyte infusion (G-DLI) has been shown to downregulate type-1 immunity in a heart transplant model, we examined influences of G-DLI on tumor growth in immunosuppressed hosts. F344 rats were treated with tacrolimus (8 mg/kg i.m.) and syngeneic colon adenocarcinoma RCN-9 cells (3 x 10(6)) were inoculated subcutaneously. For G-DLI, allogeneic DA rats were pretreated with rhG-CSF (250 microg/kg, days -5 to 0) and isolated leukocytes or sorted CD8(+) cells were injected intravenously to the hosts on day 0. Tumors in tacrolimus-treated hosts continuously grew over 5 weeks. G-DLI of 100 x 10(6) leukocytes attenuated tumor growth rate while direct host pretreatment with rhG-CSF did not. Notably, G-DLI of 10 x 10(6) CD8(+) cells blocked tumor expansion after day 14. Tacrolimus induced inhibition of lymphocyte infiltration into tumors was recovered by the G DLIs. Flow cytometry showed no detectable donor-type T cells in the tumor and circulation. Quantification of intratumor transcription levels using reverse transcription-real-time polymerase chain reaction showed recovery from tacrolimus induced downregulation of interleukin-4 but not interferon-gamma levels. In vivo rhG-CSF-treated CD8(+) allogeneic cells demonstrate potent anti-tumor effects by restoring type-2 immunity of immunosuppressed hosts. PMID- 16368245 TI - Activation of N-methyl-D-aspartate receptor induces a shift of drebrin distribution: disappearance from dendritic spines and appearance in dendritic shafts. AB - Drebrin is a major actin-filament-binding protein localized in mature dendritic spines. A recent in vivo immunoelectron microscopic study suggests that drebrin content at each dendritic spine is regulated by some unknown mechanisms. In the present in vitro study, we examined whether glutamate stimulation alters drebrin content in dendritic spines. Glutamate stimulation induced disappearance of drebrin immunostaining from dendritic spines but led to appearance of drebrin immunostaining in dendritic shafts and somata. The glutamate-induced shift of drebrin immunostaining was blocked by an NMDA receptor antagonist. Immunoblot analyses showed that both the total and the cytosolic drebrin remained unchanged and revealed that the drebrin shift was not due to drebrin degradation. These findings indicate that NMDA receptor activation induces a shift in subcellular distribution of drebrin associated with actin filaments, and that the shift might be a molecular basis for actin reorganization accompanied with synaptic plasticity. PMID- 16368246 TI - Electromyographic instantaneous amplitude and instantaneous mean power frequency patterns across a range of motion during a concentric isokinetic muscle action of the biceps brachii. AB - The purpose of this study was to examine the electromyographic (EMG) instantaneous amplitude (IA) and instantaneous mean power frequency (IMPF) patterns for the biceps brachii muscle across a range of motion during maximal and submaximal concentric isokinetic muscle actions of the forearm flexors. Ten adults (mean +/- SD age = 22.0 +/- 3.4 years) performed a maximal and a submaximal [20% peak torque (PT)] concentric isokinetic forearm flexion muscle action at a velocity of 30 degrees s(-1). The surface EMG signal was detected from the biceps brachii muscle with a bipolar electrode arrangement, and the EMG IA and IMPF versus time relationships were examined for each subject using first- and second-order polynomial regression models. The results indicated that there were no consistent patterns between subjects for EMG IA or IMPF with increases in torque across the range of motion. Some of the potential nonphysiological factors that could influence the amplitude and/or frequency contents of the surface EMG signal during a dynamic muscle action include movement of the muscle fibers and innervation zone beneath the skin surface, as well as changes in muscle fiber length and the thickness of the tissue layer between the muscle and the recording electrodes. These factors may affect the EMG IA and IMPF patterns differently for each subject, thereby increasing the difficulty of drawing any general conclusions regarding the motor control strategies that increase torque across a range of motion. PMID- 16368247 TI - Neural correlates of perceptual contributions to nondeclarative memory for faces. AB - Face priming is a nondeclarative memory phenomenon that can be observed when recognition is facilitated for a recently encountered face. This data-driven form of priming is distinct from conceptually driven priming. Moreover, it includes two dissociable components, the facilitated access to pre-existing representations and facilitation in perceptual processing of faces. In the present study, we measured neural correlates of perceptual contributions to face priming with event-related brain potentials. Faces appeared two times (separated by 7-17 s), while participants discriminated familiar from unfamiliar faces. Half of the initial face stimuli were inverted, thereby disrupting perceptual face processing and making possible an assessment of perceptual contributions to face priming. Whereas none of the brain waves previously linked to perceptual processing of faces showed indications of priming, such effects were observed between 200 and 600 ms at left occipito-parieto-temporal recording sites. This electrical activity was present for both unfamiliar and familiar faces. The scalp topography of this effect was consistent with sources within the temporal and occipital cortices of the left hemisphere (based on a LORETA source localization). These findings suggest that priming of perceptual face processing is subserved by prolonged neural activity from 200 to 600 ms primarily in the left hemisphere. We propose that this priming reflects facilitated selection based on second-order relations among facial features. PMID- 16368248 TI - MEG source localization under multiple constraints: an extended Bayesian framework. AB - To use Electroencephalography (EEG) and Magnetoencephalography (MEG) as functional brain 3D imaging techniques, identifiable distributed source models are required. The reconstruction of EEG/MEG sources rests on inverting these models and is ill-posed because the solution does not depend continuously on the data and there is no unique solution in the absence of prior information or constraints. We have described a general framework that can account for several priors in a common inverse solution. An empirical Bayesian framework based on hierarchical linear models was proposed for the analysis of functional neuroimaging data [Friston, K., Penny, W., Phillips, C., Kiebel, S., Hinton, G., Ashburner, J., 2002. Classical and Bayesian inference in neuroimaging: theory. NeuroImage 16, 465-483] and was evaluated recently in the context of EEG [Phillips, C., Mattout, J., Rugg, M.D., Maquet, P., Friston, K., 2005. An empirical Bayesian solution to the source reconstruction problem in EEG. NeuroImage 24, 997-1011]. The approach consists of estimating the expected source distribution and its conditional variance that is constrained by an empirically determined mixture of prior variance components. Estimation uses Expectation Maximization (EM) to give the Restricted Maximum Likelihood (ReML) estimate of the variance components (in terms of hyperparameters) and the Maximum A Posteriori (MAP) estimate of the source parameters. In this paper, we extend the framework to compare different combinations of priors, using a second level of inference based on Bayesian model selection. Using Monte-Carlo simulations, ReML is first compared to a classic Weighted Minimum Norm (WMN) solution under a single constraint. Then, the ReML estimates are evaluated using various combinations of priors. Both standard criterion and ROC-based measures were used to assess localization and detection performance. The empirical Bayes approach proved useful as: (1) ReML was significantly better than WMN for single priors; (2) valid location priors improved ReML source localization; (3) invalid location priors did not significantly impair performance. Finally, we show how model selection, using the log-evidence, can be used to select the best combination of priors. This enables a global strategy for multiple prior-based regularization of the MEG/EEG source reconstruction. PMID- 16368249 TI - Beyond threat: amygdala reactivity across multiple expressions of facial affect. AB - The amygdala has been consistently isolated as a key neural substrate for processing facial displays of affect. Recent evidence from human lesion and functional neuroimaging studies have begun to challenge the notion that the amygdala is reserved for signals of threat (fear/anger). We performed a 4 T fMRI study in which 20 subjects viewed a contemporary set of photographs displaying 6 different facial expressions (fearful, disgusted, angry, sad, neutral, happy) while performing a task with minimal cognitive demand. Across subjects, the left amygdala was activated by each face condition separately, and its response was not selective for any particular emotion category. These results challenge the notion that the amygdala has a specialized role in processing certain emotions and suggest that the amygdala may have a more general-purpose function in processing salient information from faces. PMID- 16368250 TI - A test of morphological hypotheses for tribal and subtribal relationships of Aphidinae (Insecta: Hemiptera: Aphididae) using DNA sequences. AB - Aphidinae is the most diverse major lineage of aphids (Aphididae). Aphidinae currently dominate the temperate, northern-hemisphere fauna, but only since the late Tertiary, and few species are native to the southern hemisphere. The success of Aphidinae may be linked to the evolution of an unusual life cycle, host alternation. The classification and phylogeny of Aphidinae have been controversial; schemes based on morphology have been confounded by widespread homoplasy. Here we present the first phylogenetic study of higher-level Aphidinae relationships based on molecular data (elongation factor-1alpha, leucine tRNA, and cytochrome oxidase II sequences). Analyses supported the monophyly of Aphidini and its subtribes, Aphidina and Ropalosiphina, but revealed novel relationships concerning Pterocommatini and Macrosiphini, with the former nested within the latter tribe as the sister to Cavariella. Several relationships within Pterocommatini + Macrosiphini corresponded better with host-plant affiliations than with aphid classification. Overall, relationships found here challenge several traditional views of Aphidinae evolution: they suggest more than one origin of host alternation in the family, and they question the assumption that Aphidinae originated in the northern hemisphere. PMID- 16368251 TI - Long-term outcome after temporal lobe surgery--prediction of late worsening of seizure control. AB - We analyzed possible predictors of late worsening of seizure control in 94 adult patients who had anterior temporal lobectomy (ATL) from the Epilepsy Center of the National Institute of Psychiatry and Neurology, Budapest between 1985 and 2001. We evaluated data regarding epilepsy, presurgical evaluation, pre- and postoperative EEG, structural imaging, histology and operative complications. The mean follow-up was 6.1 years (range: 2-17 years). The outcome was measured as Engel class, the time to the first seizure and the longest seizure free period. Multiple regression analysis was used to assess predictors. Seizure free outcome was achieved in 72% of the patients 1-year after surgery. Eighty-seven percent of them remained seizure free at the second year of follow-up, 74% at the fifth, and 67% at the tenth year of follow-up. After 2 years of follow-up improvement was present in 3%, worsening in 18% of the patients. Factors associated with long term worsening were: postoperative ipsilateral EEG spikes over the resected side, preoperative bilateral interictal discharges, cortical dysplasia of Taylor's type, and ictal contralateral propagation. In these patients, even in seizure free state, therapy reduction might be inappropriate. PMID- 16368252 TI - Downregulation of inhibitor of apoptosis proteins in apoptotic human chondrocytes treated with tumor necrosis factor-alpha and actinomycin D. AB - OBJECTIVE: Apoptosis of chondrocytes plays a pivotal role in cartilage degeneration. Tumor necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine and has been assumed to cause the degradation of human cartilage. To investigate the mechanisms of TNF-alpha-mediated apoptosis of human chondrocytes from a point of view of the balance between the caspase-cascade and the expression of inhibitor of apoptosis proteins (IAPs), although both of them are induced with TNF-signals. METHODS: The expression of TNF-receptors (TNF-Rs) in normal human articular chondrocyte (NHAC-kn) was examined with immunocytochemistry. Subconfluent cultures of NHAC-kn were tested with TNF-alpha and/or actinomycin D (actD), and the induction of apoptosis was evaluated by the frequency of apoptotic cells visualized with nuclear staining using Hoechst 33342. The activation of caspases and the expression of IAPs were examined with Western blot analyses. RESULTS: NHAC-kn expressed TNF-R1 and -R2. When NHAC-kn was treated with TNF-alpha (10 ng/ml) and actD (0.2 microg/ml) for 24 h, the frequency of apoptotic cells increased to more than 25%. TNF-alpha alone, however, induced the apoptosis insufficiently (up to 8.3%), even when used at the concentration of 100 ng/ml for 48 h. In apoptotic human chondrocytes induced with TNF-alpha (10 ng/ml) and actD (0.2 microg/ml), the caspase-3, -8, and -9 were activated and the protein expression of XIAP and c-IAP1 decreased. CONCLUSIONS: In apoptotic human chondrocytes induced with TNF-alpha and actD, the balance between caspase activation and IAPs' expression lay with the executioner caspase (caspase-3) and led to decreased expression of XIAP and c-IAP1. PMID- 16368253 TI - Evolving experience of percutaneous management of type B aortic dissection. PMID- 16368254 TI - Molecular basis for the enantioselective binding of a novel class of cytochrome bc1 complex inhibitors. AB - The recently solved co-crystal structures of mitochondrial cytochrome bc1 complex with inhibitors have provided an important structural framework for the elucidation of modes of binding of various bc1 complex inhibitors. N-Phenyl triazolones, a novel class of bc1 complex ubiquinol oxidation (Qo)-site inhibitors, were found to exhibit atropisomerism; in few cases, the atropisomers were resolved and shown to express different biological activities. However, the underlying mechanism for such differential binding of the enantiomers to bc1 complex is unknown. Here molecular docking is used to examine the binding modes of the N-phenyl triazolones fungicides. Our docking studies allow the molecular basis for the enantioselective binding of atropisomeric triazolones to be elucidated. Furthermore, the mode of binding of azoxystrobin has also been clarified. PMID- 16368255 TI - 'Wound cat'. AB - 'Wound man' refers to a stylised diagram used in early medical textbooks to illustrate the various injuries that the human body can sustain in battle. We have adapted this concept to create 'Wound cat', as a way to emphasise the type and location of injuries cats may inflict on one another during combat. We have further extended this concept to include wounds that may result from interactions with rodents and snakes. It is hoped that our 'Wound cat' concept will assist less experienced clinicians locate sites of cat bites and scratches, and to recognise why certain infections arise in particular stereotyped locations. In addition, this approach should assist veterinarians in determining which pathogens are most likely to be involved in wounds located in a given anatomical region. PMID- 16368256 TI - Clinical course in Parkinson's disease with elevated homocysteine. AB - Elevated homocysteine (Hcy), prevalent in Parkinson's disease (PD), is potentially a modifiable risk factor for neurologic deterioration. We measured cognitive, affective and motor changes over 2 years in a cohort of people with early PD. Subjects whose Hcy had been elevated (>14 micromol/L, n = 31) at baseline were compared with the rest (n = 66). Overall progression in 2 years did not significantly differ (p = 0.20). Four subjects with elevated and one with normal Hcy had died (p = 0.03). We conclude that hyperhomocysteinemia does not predict significantly worse progression over 2 years in early PD. The data raised the possibility of higher mortality, but the number of deaths was small. PMID- 16368257 TI - Atypical parkinsonism and SCA8. PMID- 16368258 TI - Cortical and amygdalar Lewy body burden in Parkinson's disease patients with visual hallucinations. AB - Visual hallucinations (VH) are among the most common non-motor complications of Parkinson's disease (PD). A few studies on their etiopathogenesis have suggested involvement of cortical and amygdalar areas. In order to investigate the possible association between extranigral Lewy Body (LB) distribution across cortical and amygdalar regions and the presence of VH in PD brain donors, we conducted a clinico-pathological comparison of 10 PD patients with VH vs 10 closely matched PD patients without VH. The LB burden was significantly higher across the amygdala and the frontal, temporal and parietal cortical areas in patients with VH. Although our results suggest significant extranigral involvement, the precise etiopathologic mechanisms responsible for the development of VH need further clarification. PMID- 16368260 TI - The Intelligent Design controversy: lessons from psychology and education. PMID- 16368261 TI - Identification and intra-specific variability analysis of secreted and membrane bound proteins from Echinococcus granulosus. AB - Echinococcus granulosus, the etiological agent of cystic hydatid disease, exists as a series of strains or genotypes, differing in biological features. Many of the secreted and membrane-bound proteins (S/M) from helminth parasites are involved in the host-parasite interplay and constitute potential targets for diagnosis, anti-parasitic drugs and vaccines. A number of E. granulosus S/M proteins were identified using the signal sequence trap technique. Six out of seven cDNA fragments of these newly identified proteins showed nucleotide and amino acid sequence variation. Inter-strain variation was reported for other characterized S/M proteins as the vaccine target EG95 and the major hydatid cyst fluid antigen, Antigen B (AgB). AgB is highly polymorphic, 101 different sequences related to AgB were reported so far and were grouped in 5 genes (EgB1 EgB5) and one pseudogene (EgB2p) exclusive of G5, G6/G7 genotypes. The significance of AgB polymorphism and possible consequences in diagnostic performance are discussed. The diagnostic value of the new protein variants detected in E. granulosus strains could be determined through standardized inter laboratory studies as the recently done by the South American Network for Hydatid Serology. PMID- 16368263 TI - Diagnosing entrapment neuropathies: probes and magnets instead of electrodes and needles? PMID- 16368262 TI - Congenital cytomegalovirus infection following first trimester maternal infection: symptoms at birth and outcome. AB - BACKGROUND: The relationship between gestational age at time of maternal cytomegalovirus (CMV) infection and outcome of fetal infection is not well defined because the timing of maternal infection is usually not known. OBJECTIVE: To determine whether congenital cytomegalovirus (CMV) infection following primary maternal infection during the first trimester of pregnancy is more likely to lead to central nervous system (CNS) sequelae than fetal infection due to maternal infection later in pregnancy. STUDY DESIGN: Using serum collected during pregnancy from mothers of newborns with congenital CMV infection, maternal infection was categorized as first trimester (<13 weeks) or later based on dates and results of IgG and IgM assays for CMV antibody. Outcome of congenital CMV infection was assessed by longitudinal fotlow-up of the infected cohort. RESULTS: Sensorineural hearing loss was found in 8/34 (24%) of children in the first trimester group, compared with 1/40 (2.5%) in the later infection group (P=0.01, relative risk, 9.6). Considering any CNS sequela (hearing loss, mental retardation, cerebral palsy, seizures, chorioretinitis) 11/34 (32%) first trimester cases were affected compared with 6/40 (15%) in the later infection group (P=0.07, relative risk 2.2). None of the later group had more than one sequela, compared with 4 (12%) of the first trimester group (P=0.04). CONCLUSIONS: Children with congenital CMV infection following first trimester maternal infection are more likely to have CNS sequelae, especially sensorineural hearing loss, than are those whose mothers were infected later in pregnancy. However, some degree of CNS impairment can follow even late gestational infection. PMID- 16368264 TI - Patient choice in cancer follow-up. PMID- 16368265 TI - Gaboxadol--a new awakening in sleep. AB - Drugs that enhance synaptic gamma-aminobutyric acid (GABA)ergic neurotransmission are widely utilized in the clinical setting. Barbiturates and benzodiazepine receptor agonists, for example, both potentiate an inhibitory chloride conductance through GABA-gated channels, and thereby achieve their sedative hypnotic effects. The primary locus of action of these agents, and indeed most neuroactive drugs, is the postsynaptic junction. By contrast, gaboxadol, a selective extrasynaptic GABA receptor agonist and late-stage investigational treatment for insomnia, acts on a unique delta-containing GABAA receptor subtype found exclusively outside of the synapse. Although the mechanistic details of extrasynaptic neurotransmission remain to be fully established, it is now clear that these receptors demonstrate unique pharmacological, biophysical and electrophysiological properties. Importantly, the delta-containing GABAA receptor subtype activated by gaboxadol is highly expressed in the thalamus, where it might behave as a 'gain control' (independently controlling the strength of signals) in the corticothalamic pathways that govern sleep-relevant neuronal oscillations. This unique mechanism has contributed to our increased understanding of sleep mechanisms, and targeting of this system offers potential advantages over existing insomnia treatments. PMID- 16368266 TI - Mechanism of action of memantine. AB - Memantine is a clinically useful drug in many neurological disorders, including Alzheimer's disease. The principal mechanism of action of memantine is believed to be the blockade of current flow through channels of N-methyl-d-aspartate (NMDA) receptors--a glutamate receptor subfamily broadly involved in brain function. Surprisingly, other drugs that block NMDA receptor channels, such as ketamine, exhibit serious deleterious effects. The unusual therapeutic utility of memantine probably results from inhibitory mechanisms shared with ketamine, combined with actions specific to memantine. These potentially important differences between memantine and ketamine include effects on gating of blocked channels and binding of memantine to two sites on NMDA receptors. Because modulation of NMDA receptor activity can increase or decrease excitability of neuronal circuits, subtle differences in the mechanisms of action of NMDA receptor antagonists can strongly impact on their clinical effects. PMID- 16368267 TI - Unravelling the brain targets of gamma-hydroxybutyric acid. AB - Gamma-hydroxybutyric acid (GHB) is a naturally occurring gamma-aminobutyric acid (GABA) metabolite that has been proposed as a neurotransmitter/neuromodulator that acts via its own receptor (GHBR). Its exogenous administration, however, elicits central nervous system-dependent effects (e.g. memory impairment, increase in sleep stages 3 and 4, dependence, seizures and coma) that are mostly mediated by GABAB receptors. The past few years have seen important developments in our understanding of GHB neurobiology: a putative GHBR has been cloned; a transgenic model of GHB aciduria has been developed; GABAB receptor knockout mice and novel GHB analogs have helped to characterize the vast majority of exogenous GHB actions mediated by GABAB receptors; and some of the cellular mechanisms underlying the dependence/abuse properties of GHB, and its ability to elicit absence seizures and an increase in sleep stages 3 and 4, have been clarified. Nevertheless, the physiological significance of a brain GHB signaling pathway is still unknown, and there is an urgent need for a well-validated functional assay for GHBRs. Moreover, as GHB can also be metabolized to GABA, it remains to be seen whether the many GABAB receptor-mediated actions of GHB are caused by GHB itself acting directly on GABAB receptors or by a GHB-derived GABA pool (or both). PMID- 16368268 TI - Glutamate-based therapeutic approaches: allosteric modulators of metabotropic glutamate receptors. AB - Metabotropic glutamate receptors (mGluRs) have been proposed as novel targets for the treatment of a variety of disorders. Recently, highly selective allosteric modulators of the mGluRs have been developed by several groups. These allosteric compounds provide an unprecedented degree of selectivity for individual mGluRs, allowing for more detailed functional studies on the roles of these receptors. Furthermore, the allosteric approach avoids many of the hurdles associated with the development of direct agonists as drugs, and provides a clear path forward for clinical proof-of-concept studies. Currently, both positive allosteric modulators of mGluR2 and negative allosteric modulators of mGluR5 hold promise as novel anxiolytics, and positive allosteric modulators of mGluR4 appear to be an exciting new target for the treatment of Parkinson's disease. PMID- 16368269 TI - Glutamate-based therapeutic approaches: targeting the glutamate transport system. AB - Although the ionotropic and metabotropic receptors for synaptically released glutamate have been extensively mined in the pursuit of novel therapeutic agents for a diverse array of central nervous system disorders, pursuit of the transport proteins--or excitatory amino acid transporters (EAATs)--toward a similar end has been a road much less travelled. Recent progress has seen the use of cloned EAAT subtypes to develop transporter inhibitors with improved subtype selectivity, providing important tools for elucidating the precise contribution of each transporter subtype to the regulation of extracellular glutamate homeostasis. In addition, momentum has been gained with the discovery of compounds capable of upregulating the activity of the predominant forebrain glutamate transporter, EAAT2. PMID- 16368270 TI - A phenotype based approach for the immune monitoring of NY-ESO-1-specific CD4+ T cell responses in cancer patients. AB - Because of its frequent expression in tumors and spontaneous immunogenicity in advanced cancer patients, NY-ESO-1 is presently viewed as a prototype tumor antigen for the development of cancer vaccines. A prerequisite for the analysis of NY-ESO-1-specific T cell responses in vaccinated patients is the assessment of the complete T cell repertoire available for the antigen. Here, we have assessed frequency and fine specificity of CD4+ T cells reactive against NY-ESO-1-derived sequences in circulating lymphocytes from cancer patients with spontaneous responses to the antigen. We found that, relative to healthy donors, this frequency was only moderately increased in cancer patients. The reactivity of these cells, however, was directed against the same immunodominant regions previously identified for healthy donors. On account of these data, we developed an approach for the immune monitoring of NY-ESO-1-specific CD4+ T cell responses based on the assessment of CD4+ T cell populations of defined phenotype. Using this approach, a similar frequency of NY-ESO-1-specific CD4+ T cells was found among naive T cells of healthy donors and cancer patients. In contrast, among antigen-experienced T cells, NY-ESO-1-specific CD4+ T cells were exclusively detectable in cancer patients. We anticipate that this phenotype-based approach will be useful for the immune monitoring of vaccine-induced responses in vaccination trials using NY-ESO-1 as well as other tumor antigens. PMID- 16368271 TI - Prevention of ovarian cancer. AB - Ovarian cancer is the leading cause of death from gynaecological malignancy. The incidence is high in the Western world. The incidence of ovarian cancer is reduced by pregnancy, lactation, the oral contraceptive pill and tubal ligation. Lifestyle factors are important in the aetiology of ovarian cancer and current evidence suggests the risk can be reduced by eating a diet rich in fruit and vegetables, taking regular exercise, avoiding smoking, avoiding being overweight and avoiding long-term use of hormonal replacement therapy (HRT). Familial ovarian cancer is responsible for about 10% of ovarian cancer cases. Strategies available to high-risk women include screening (covered elsewhere) and prophylactic salpingo-oophorectomy. The precise role of chemoprevention for high risk women in the form of the oral contraceptive pill is unclear. PMID- 16368272 TI - RNA as a source of transposase for Sleeping Beauty-mediated gene insertion and expression in somatic cells and tissues. AB - Sleeping Beauty (SB) is a DNA transposon capable of mediating gene insertion and long-term expression in vertebrate cells when co-delivered with a source of transposase. In all previous reports of SB-mediated gene insertion in somatic cells, the transposase component has been provided by expression of a co delivered DNA molecule that has the potential for integration into the host cell genome. Integration and continued expression of a gene encoding SB transposase could be problematic if it led to transposon re-mobilization and reintegration. We addressed this potential problem by supplying the transposase-encoding molecule in the form of mRNA. We show that transposase-encoding mRNA can effectively mediate transposition in vitro in HT1080 cells and in vivo in mouse liver following co-delivery with a recoverable transposon or with a luciferase transposon. We conclude that in vitro-transcribed mRNA can be used as an effective source of transposase for SB-mediated transposition in mammalian cells and tissues. PMID- 16368273 TI - AAV transcytosis through barrier epithelia and endothelium. AB - To transduce efficiently barrier epithelia such as the lung is the goal of several gene therapy applications. However, experiments with AAV-2 suggest that transduction is limited in this type of barrier epithelia. In contrast, other serotypes of AAV transduce barrier epithelia and exhibit broad dissemination throughout the tissue. Transcytosis is a process by which proteins and pathogens overcome barrier layers to reach the opposite cell surface. To understand better the entry pathway of AAV particles and their ability to penetrate barrier epithelia, we tested the hypothesis that the limited transduction of some barrier epithelia in vitro or the spread of some AAV serotypes through tissue in vivo is due to transcytosis. Our experiments demonstrate that dependoviruses can penetrate barrier cells by transcytosis. The process is rapid as well as serotype and cell-type specific and can be blocked by neutralizing antibodies, temperature, or chemical inhibitors of transcytosis. The particles isolated following apical-to-basolateral transport are still encapsulated and they can transduce permissive cell lines in vitro. Furthermore, the entry pathway used by AAV-5 for transcytosis appears to be independent of the one used for transduction. Importantly, inhibition of virus transcytosis results in a dramatic increase in intracellular vector and transduction. PMID- 16368274 TI - Happy seventh birthday, Epilepsy and behavior. PMID- 16368275 TI - Determining empirically based self-reported cognitive change: development of reliable change indices and standardized regression-based change norms for the multiple abilities self-report questionnaire in an epilepsy sample. AB - PURPOSE: Reliable change indices (RCIs) and standardized regression-based (SRB) change score norms were calculated for a measure of self-reported cognitive function, the Multiple Abilities Self-Report Questionnaire (MASQ), in patients with complex partial seizures. Establishment of such standardized change scores could be useful in determining the magnitude and direction of self-appraised cognitive change after epilepsy surgery or other treatment interventions. The primary study objective was to calculate RCI and SRB values for the MASQ. A secondary objective was to report SRB change scores in patients who had undergone anterior temporal lobectomy (ATL) and to assess relationships between self reported cognitive change, seizure outcome, objective memory test performance, and mood. METHODS: The MASQ was administered to 36 patients with complex partial seizures on two occasions (mean test-retest interval, 6 months). This group did not have major psychopathology and were on stable antiepileptic drugs. RCI and SRB change scores were calculated. Adjustments for baseline ratings, age, education, gender, age at seizure onset, and seizure duration were made with the SRB method. A confidence interval cutoff score (90% level) was calculated for the five MASQ cognitive domains (Language, Visual Perception, Verbal Memory, Visual Spatial Memory, Attention/Concentration). MASQ SRB scores were computed for a second sample of 50 patients who had undergone ATL. RESULTS: Test-retest reliabilities for the MASQ domains ranged from a low of 0.63 (Attention/Concentration) to a high of 0.87 (total score). Baseline MASQ score was the single largest contributor to the regression equations. Left and right ATL groups demonstrated similar magnitudes of self-reported cognitive change across all five MASQ domains. Individual base rate change distributions were similar across four of the five domains. with a higher proportion of right ATL patients reporting worsening attention function. Both postoperative mood and SRB based verbal memory outcome were significantly correlated to self-reported cognitive change in the patients who had undergone ATL. CONCLUSIONS: SRB methodology provides a standardized technique with which to establish patient perception of cognitive change and may be of use when examining change across individual- and group-level ratings of cognitive functioning in clinical and research settings. These techniques also provide a common metric for direct comparison between subjective self-report ratings of cognitive function and objective cognitive test instruments. PMID- 16368276 TI - Sumoylation of PCNA: Wrestling with recombination at stalled replication forks. AB - Post-replication repair encompassses error-prone and error-free processes for bypassing lesions encountered during DNA replication. In Saccharomyces cerevisiae, proteins acting in the Rad6-dependent pathway are required to channel lesions into these pathways. Until recently there was little information as to how this channelling was regulated. However, several recent papers, and in particular from the Jentsch and Ulrich groups have provided striking insights into the role of modified forms of PCNA in these events [C. Hoege, B. Pfander, G.L. Moldovan, G. Pyrowolakis, S. Jentsch, RAD6-dependent DNA repair is linked to modification of PCNA by ubiquitin and SUMO, Nature 419 (2002) 135-141; P. Stelter, H.D. Ulrich, Control of spontaneous and damage-induced mutagenesis by SUMO and ubiquitin conjugation, Nature 425 (2003) 188-191; B. Pfander, G.L. Moldovan, M. Sacher, C. Hoege, S. Jentsch, SUMO-modified PCNA recruits Srs2 to prevent recombination during S phase, Nature 436 (2005) 428-433; E. Papouli, S. Chen, A.A. Davies, D. Huttner, L. Krejci, P. Sung, H.D. Ulrich, Crosstalk between SUMO and ubiquitin on PCNA is mediated by recruitment of the helicase Srs2p, Mol. Cell. 19 (2005) 123-133]. In particular they have shown that mono-ubiquitinated PCNA directs translesion synthesis via DNA polymerases with low stringency, and that polyubiquitinated PCNA is associated with error-free avoidance of lesions. Recent data have shown that the role of small ubiquitin-like modifier (SUMO) modification of PCNA is not an event that occurs merely in the absence of ubiquitination, rather it serves to recruit Srs2 to replication forks in order to inhibit recombination. The implications of these findings for post-replication repair in S. cerevisiae and other eukaryotes are discussed. PMID- 16368277 TI - The role of neonatal networks. AB - A neonatal network is a collaboration involving more than one clinical site where a common protocol is used for a randomized trial, observational study, or quality improvement project. Because the prevalence of important outcomes (e.g., death or neurodevelopmental impairment) is low among neonates, multi-site studies have been an essential contributor to the identification of interventions for improving the health of neonates. In this review we summarize key aspects of the conduct of multi-site studies and describe some of the prominent accomplishments and contributions of neonatal networks. Important contributions of randomized trials and observational studies include the identification, and assessment, of potential strategies for prevention of cerebral palsy, chronic lung disease, and blindness due to retinopathy of prematurity. In addition, quality improvement projects by neonatal networks have greatly facilitated the dissemination of information about, and implementation of, evidence-based interventions. PMID- 16368278 TI - Fetal origins of adult disease. AB - The term 'fetal origins of adult disease' was coined on the basis of the inverse association between low birth weight and blood pressure, adult-onset diabetes, coronary heart disease, and stroke seen in numerous epidemiological studies. However, it seems unlikely that birth weight is involved in causal pathways underlying these observations, and if it were then the significance to public health of these findings is very limited because of our inability to modify birth weight to a relevant extent in humans. There has been a major focus on maternal nutrition. Despite evidence that experimental manipulation of maternal nutrition in animals influences offspring birth weight and programme measures related to cardiovascular disease, human studies in general provide limited and unconvincing evidence that differences in maternal macronutrient intake are important. Nevertheless there is a need to understand the underlying causal pathways, and the utility of studies of twins and possible mechanisms are discussed. PMID- 16368279 TI - The effects of local platelet rich plasma delivery on diabetic fracture healing. AB - Several studies have documented that diabetes impairs bone healing clinically and experimentally. The percutaneous delivery of platelet rich plasma (PRP) was used in the diabetic BB Wistar femur fracture model to investigate the use of PRP as a concentrated source of critical early growth factors on bone healing. PRP delivery at the fracture site normalized the early (cellular proliferation and chondrogenesis) parameters while improving the late (mechanical strength) parameters of diabetic fracture healing. These results suggest a role for PRP in mediating diabetic fracture healing and potentially other high risk fractures. PMID- 16368280 TI - Cost-effectiveness analysis (CEA) health economic point of view. PMID- 16368282 TI - The cost-effectiveness analysis (CEA) is dependent on the quality of the NEPHRIC study. PMID- 16368284 TI - The effects of oral anticoagulants in patients with peripheral arterial disease: rationale, design, and baseline characteristics of the Warfarin and Antiplatelet Vascular Evaluation (WAVE) trial, including a meta-analysis of trials. AB - AIMS: Patients with peripheral arterial disease (PAD) are at a high risk for cardiovascular (CV) morbidity and mortality even when treated with antiplatelet therapy. We present the rationale (including a meta-analysis of relevant trials), design, and baseline characteristics of the WAVE trial evaluating oral anticoagulants (OAC) in PAD. METHODS AND RESULTS: Nine trials involving 4889 patients with PAD evaluating OAC have conflicting results. Combining the data, it appears that OAC may reduce mortality and graft occlusion but increase major bleeding compared with no treatment. Compared with aspirin, OAC do not appear to reduce mortality (odds ratio [OR] = 1.04, 95% CI 0.55-1.29), although the CI are wide, or graft occlusion (OR = 0.91, 95% CI 0.77-1.06), and major bleeding is increased (OR = 1.96, 95% CI 1.43-2.69). Compared with aspirin, OAC used together with aspirin appears to increase mortality (OR = 1.57, 95% CI 1.16-2.12); may reduce graft occlusion (OR = 0.84, 95% CI 0.62-1.12), and major bleeding is increased (OR = 2.13, 95% CI 1.27-3.57). To further clarify the efficacy and safety profile of OAC in patients with PAD, we initiated the WAVE trial in which patients with PAD are randomized to receive OAC (target international normalizing ratio 2-3) plus antiplatelet therapy or antiplatelet therapy alone. Patients are treated for a minimum of 2.5 years and a maximum of 3.5 years. The co-primary efficacy outcomes are (1) CV death, MI, and stroke; and (2) CV death, MI, stroke, and acute limb or coronary ischemia requiring urgent intervention. The baseline characteristics of the study population confirm that patients with PAD represent a high-risk group. CONCLUSIONS: The results of previous randomized trials evaluating OAC in patients with PAD (who represent a group at a high risk for thrombotic events) are inconclusive. WAVE is a large, international, randomized clinical trial designed to determine if moderate levels of oral anticoagulation (international normalizing ratio 2-3) improve upon antiplatelet therapy alone. PMID- 16368285 TI - Unraveling the spectrum of left bundle branch block in acute myocardial infarction: insights from the Assessment of the Safety and Efficacy of a New Thrombolytic (ASSENT 2 and 3) trials. AB - BACKGROUND: Left bundle branch block (LBBB) complicates the diagnosis of acute myocardial infarction (AMI). The Sgarbossa criteria were developed from GUSTO I to surmount this diagnostic challenge but have not been prospectively validated in a large population with presumed AMI. We evaluated their utility in the diagnosis and risk stratification of AMI patients in ASSENT 2 & 3. METHODS: Baseline electrocardiograms (ECG) of LBBB patients were scored using Sgarbossa's criteria (0-10) by 2 readers blinded to the CK/CK-MB data and clinical outcomes; 267 (1.2%) patients had LBBB on their baseline ECG. RESULTS: Among 253 LBBB patients with available peak CK/CK-MB data, 158 (62.5%) had peak CK/CK-MB levels > 2x ULN, thereby qualifying for the diagnosis of AMI. A Sgarbossa score of 3 was shown in 48.7% of LBBB patients with elevated CK/CK-MB versus in 12.6% of those without a CK/CK-MB rise (P < .001). Patients with higher Sgarbossa scores, ie, 3, had a higher mortality compared with those with a score < 3, (23.5% vs 7.7% at 30 days P < .001; and 33.7% vs 20.2% at 1 year, P < .001, respectively). CONCLUSIONS: Our findings validate the utility of Sgarbossa criteria for diagnosing AMI in the setting of LBBB. These criteria provide a simple and practical diagnostic approach to risk stratify this diagnostically challenging high-risk group and optimize risk-benefit of acute therapy. PMID- 16368286 TI - Elevated creatine kinase-MB with normal creatine kinase predicts worse outcomes in patients with acute coronary syndromes: results from 4 large clinical trials. AB - BACKGROUND: The degree to which elevated creatine kinase (CK)-MB in the presence of normal CK is predictive of outcome is not well understood despite having been studied for decades. This analysis examined whether normal CK with elevated CK-MB in patients with non-ST-segment elevation acute coronary syndrome (NSTE ACS) is an independent predictor of worse outcomes. A concomitant goal was to contribute insight to the debate over how patients with NSTE ACS should be managed. METHODS: Data for 25,960 patients from the GUSTO IIb, PARAGON A and B, and PURSUIT trials were analyzed. Of these patients, 6402 were excluded from primary analysis because of missing (unmeasured) biomarkers. Patients with complete laboratory data (n = 19,558) were grouped by CK and CK-MB results. To confirm the primary analysis results, data from patients with missing biomarkers were used in an imputation model. RESULTS: Patients were categorized in 1 of 4 groups: normal CK + normal CK-MB; normal CK + elevated CK-MB; elevated CK + normal CK-MB; or elevated CK + elevated CK-MB. For the primary outcome, 180-day death, or myocardial infarction, Kaplan-Meier estimates were 14.9%, 20.8%, 14.5%, and 18.2%, respectively. Regardless of total CK, elevated CK-MB was associated with a 25% to 49% increased relative risk of worse outcomes. Findings from the analyses were verified by the multivariable model. CONCLUSIONS: CK-MB remains a reliable marker for myocardial necrosis and a strong predictor of worse prognosis. All patients with ACS should have CK-MB measurement to search for cardiac ischemia. Patients with elevated CK-MB should receive aggressive management commensurate with their increased risks. PMID- 16368287 TI - Combination of quantitative ST deviation and troponin elevation provides independent prognostic and therapeutic information in unstable angina and non-ST elevation myocardial infarction. AB - BACKGROUND: ST deviation and troponin are indicators of risk in unstable angina/non-ST-elevation myocardial infarction with related but distinct pathophysiology. We hypothesized that a combination of the two would offer complementary information regarding the benefit of an invasive strategy. METHODS AND RESULTS: Electrocardiogram and troponin data were gathered in 1846 patients in TACTICS-TIMI 18. Adjusting for baseline characteristics, the independent odds ratio (OR) (95% CI) for death or myocardial infarction by 6 months was 1.29 (0.78 2.13) for 0.05 to 0.09 mV ST deviation, 1.83 (1.22-2.75) for > or = 0.10 mV ST deviation, 1.69 (0.96-2.97) for troponin T 0.01 to 0.10 ng/mL, and 2.32 (1.52 3.53) for troponin T > 0.10 ng/mL. There was a significant interaction between the magnitude of ST deviation and treatment strategy (P(interaction) = .04). After adjusting for troponin and other baseline characteristics, an early invasive strategy was associated with a 27% greater reduction in the odds of death or myocardial infarction in those with 0.05 to 0.09 mV of ST deviation (OR(interaction) 0.73, 95% CI 0.27-2.02) and a 64% greater reduction in those with > or = 0.10 mV (OR(interaction) 0.36, 95% CI 0.16-0.84) compared with in those without ST deviation. CONCLUSIONS: The magnitude of ST deviation and degree of troponin elevation are graded independent predictors of outcome in unstable angina/non-ST-elevation myocardial infarction. ST deviation complements troponin elevation in selecting patients for early invasive management. PMID- 16368288 TI - Outcomes after acute coronary syndrome admission to primary versus tertiary Veterans Affairs medical centers: the Veterans Affairs Access to Cardiology study. AB - BACKGROUND: There is a concern that patients with acute coronary syndrome (ACS) admitted to primary care hospitals (without on-site cardiac procedures) may be at risk for worse outcomes compared with patients admitted to tertiary care hospitals. In addition to mortality, one way to assess patient outcomes is via health status and rehospitalization rates. We compared the health status and rehospitalization of patients with ACS admitted to primary versus tertiary care Veterans Affairs hospitals. METHODS: This was a cohort study of 2132 patients with ACS admitted to 21 Veterans Affairs hospitals (12 primary care and 9 tertiary care) from 1998 to 1999. Primary outcomes were 7-month health status as measured by the Seattle Angina Questionnaire and rehospitalization. Hierarchical multivariable regression was used to evaluate the association between admission to a primary (vs tertiary) care hospital and these outcomes. Discharge medications and 7-month cardiac procedure rates were also compared. RESULTS: There were no significant differences in discharge medication rates between primary and tertiary hospital patients. Forty-two percent of the patients admitted to a primary care hospital was transferred to a tertiary care hospital during index admission. Primary hospital patients had significantly lower 7-month rates of cardiac catheterization (36% vs 51%, P < .001) and percutaneous coronary intervention (11% vs 20%, P < .001), but there were no differences in coronary artery bypass graft surgery rates. After risk adjustment, there were no significant differences in 7-month angina frequency (odds ratio [OR] 0.98, 95% CI 0.78-1.22), physical limitation (OR 0.97, 95% CI 0.77-1.23), quality of life (OR 1.12, 95% CI 0.89-1.40), or rehospitalization (OR 1.07, 95% CI 0.54-2.14) between the 2 groups. CONCLUSIONS: These results suggest that an integrated health care system can achieve similar intermediate-term health status and rehospitalization outcomes for patients with ACS irrespective of the site of admission despite the lower rates of cardiac procedures for the primary care hospital patients. PMID- 16368289 TI - Clinical significance of calcification of the fibrous skeleton of the heart and aortosclerosis in community dwelling elderly. The Cardiovascular Health Study (CHS). AB - BACKGROUND: Mitral annular calcification (MAC), aortic annular calcification (AAC), and aortic valve sclerosis (AVS) are associated with aging, and MAC and AVS are markers of advanced atherosclerosis. No studies have examined the prevalence and the clinical relevance of all 3 forms of calcification in a single free-living elderly population. METHODS: We used 2-dimensional echocardiography to evaluate MAC, AAC, AVS and all 3 combined in 3929 participants, mean age 76 +/ 5 years, 60% women, in the Cardiovascular Health Study, a prospective community based observational study designed to assess cardiovascular disease (CVD) risk factors and outcomes in elderly persons. RESULTS: Mitral annular calcification was found in 1640 (42 %) subjects, AAC in 1710 (44 %), AVS in 2114 (54 %), and all 3 combined in 662 (17 %). The participants with these findings were older than those without them, and those with MAC had worse cardiovascular, renal, metabolic, and functional profile than those with AAC and AVS. Age-, sex-, and race-adjusted logistic regression analysis found a significant association between the 3 calcification categories and CVD, the strongest being between the combined group with congestive heart failure (odds ratio 2.04, 95% CI 1.34-3.09). In highly adjusted models, only MAC was associated with CVD, and the strength of association was related to the severity of MAC. CONCLUSIONS: In free-living elderly, MAC, AAC, and AVS are highly prevalent and are associated with CVD. Mitral annular calcification in particular has strong association with CVD, and with an adverse biomedical profile. PMID- 16368290 TI - The effects of periodontal therapy on vascular endothelial function: a pilot trial. AB - BACKGROUND: Chronic periodontal infection is associated with an increased risk of coronary heart disease. Although the mechanism responsible for the relationship between periodontal disease and cardiovascular events is not fully understood, it is hypothesized that the chronic inflammatory burden of periodontal disease may lead to impaired functioning of the vascular endothelium. METHODS: Twenty-two otherwise healthy adults with moderate to severe periodontitis who underwent complete mouth disinfection were evaluated to determine if periodontal therapy would result in improved endothelial function and a decrease in serum inflammatory markers. Subjects had measurements of periodontal disease severity, flow-mediated (endothelium-dependent), and nitroglycerin-mediated (endothelium independent) dilation of the brachial artery, serum C-reactive protein (CRP) and interleukin 6 (IL-6), and serum total and high-density lipoprotein cholesterol levels on 2 baseline visits separated by 1 month and, again, 1 month after treatment. RESULTS: There were no significant changes in clinical periodontal measures, flow-mediated dilation, nitroglycerin-mediated dilation, CRP, IL-6, total cholesterol, or high-density lipoprotein cholesterol between the repeated baseline measurements. Periodontal treatment, however, resulted in significant improvements in periodontal pocketing, flow-mediated dilation, and serum IL-6, as well as a trend toward reduction in serum CRP; there were no significant changes in nitroglycerin-mediated dilation or in cholesterol levels. CONCLUSIONS: These results represent proof of concept that improvement in endothelial function, as measured by flow-mediated dilation of the brachial artery, may be possible through near-elimination of chronic oral infection and suggest that the conduct of a larger controlled trial is justified. PMID- 16368291 TI - A validated clinical and biochemical score for the diagnosis of acute heart failure: the ProBNP Investigation of Dyspnea in the Emergency Department (PRIDE) Acute Heart Failure Score. AB - BACKGROUND: No method integrating amino-terminal pro-brain natriuretic peptide (NT-proBNP) testing with clinical assessment for the evaluation of patients with suspected acute heart failure (HF) has been described. METHODS: Amino-terminal pro-brain natriuretic peptide results and clinical factors from 599 patients with dyspnea were analyzed. The beta coefficients of the 8 independent predictors of HF were used to assign a weighted integeric score for predictor. The sum of these integers provided a diagnostic HF "score" for each patient. Receiver operating characteristic curve analysis determined the optimal cut point for the diagnosis of acute HF. The performance of the score was evaluated in the development cohort and subsequently in a patient population from a separate clinical trial of patients with dyspnea conducted in Christchurch, New Zealand. RESULTS: Eight factors comprised the score: elevated NT-proBNP (4 points), interstitial edema on chest x-ray (2 points), orthopnea (2 points), absence of fever (2 points), loop diuretic use, age > 75 years, rales, and absence of cough (all 1 point). Median scores in patients with acute HF were higher than those without acute HF (9 vs 3 points, P < .001). At a cut point of > or = 6 points, the score had a sensitivity of 96% and a specificity of 84% for the diagnosis of acute HF (P < .001). The score improved diagnostic accuracy over NT-proBNP testing alone and retained discriminative capacity in patients in whom clinical uncertainty was present. Lastly, the accuracy of the score was validated in the external data set of patients with suspected acute HF. CONCLUSION: We report a simple and accurate scoring system combining NT-proBNP testing and clinical assessment for the diagnosis or exclusion of acute HF in patients with dyspnea. PMID- 16368292 TI - Are beta-blockers needed in patients receiving spironolactone for severe chronic heart failure? An analysis of the COPERNICUS study. AB - BACKGROUND: The beneficial effects of beta-blockers and aldosterone receptor antagonists are now well established in patients with severe systolic chronic heart failure (CHF). However, it is unclear whether beta-blockers are able to provide additional benefit in patients already receiving aldosterone antagonists. We therefore examined this question in the COPERNICUS study of 2289 patients with severe CHF receiving the beta1-beta2/alpha1 blocker carvedilol compared with placebo. METHODS: Patients were divided post hoc into subgroups according to whether they were receiving spironolactone (n = 445) or not (n = 1844) at baseline. Consistency of the effect of carvedilol versus placebo was examined for these subgroups with respect to the predefined end points of all-cause mortality, death or CHF-related hospitalizations, death or cardiovascular hospitalizations, and death or all-cause hospitalizations. RESULTS: The beneficial effect of carvedilol was similar among patients who were or were not receiving spironolactone for each of the 4 efficacy measures. For all-cause mortality, the Cox model hazard ratio for carvedilol compared with placebo was 0.65 (95% CI 0.36 1.15) in patients receiving spironolactone and 0.65 (0.51-0.83) in patients not receiving spironolactone. Hazard ratios for death or all-cause hospitalization were 0.76 (0.55-1.05) versus 0.76 (0.66-0.88); for death or cardiovascular hospitalization, 0.61 (0.42-0.89) versus 0.75 (0.64-0.88); and for death or CHF hospitalization, 0.63 (0.43-0.94) versus 0.70 (0.59-0.84), in patients receiving and not receiving spironolactone, respectively. The safety and tolerability of treatment with carvedilol were also similar, regardless of background spironolactone. CONCLUSION: Carvedilol remained clinically efficacious in the COPERNICUS study of patients with severe CHF when added to background spironolactone in patients who were practically all receiving angiotensin converting enzyme inhibitor (or angiotensin II antagonist) therapy. Therefore, the use of spironolactone in patients with severe CHF does not obviate the necessity of additional treatment that interferes with the adverse effects of sympathetic activation, specifically beta-blockade. PMID- 16368293 TI - The nonspecific anti-inflammatory therapy with methotrexate for patients with chronic heart failure. AB - BACKGROUND: Inflammatory mediators play an important role in the pathogenesis of chronic heart failure (CHF). Methotrexate (MTX) is used in the treatment of inflammatory-mediated diseases (eg, rheumatoid arthritis) because it modulates the expression of numerous inflammatory cytokines. However, no studies have assessed the effects of MTX on plasma levels of inflammatory mediators in patients with CHF. METHODS: In a prospective, randomized, placebo-controlled, single-blind study, 71 patients receiving conventional treatment were randomly allocated to either MTX group (7.5 mg once a week, n = 35) or placebo group (n = 36) with a follow-up of 12 weeks. The effects of MTX on plasma cytokine expression, left ventricular ejection fraction, left ventricular end-diastolic dimension, New York Heart Association (NYHA) functional class, 6-minute walk test distance, and quality of life (QOL) were determined in patients with CHF. RESULTS: Sixty-two patients completed the study. The circulating levels of inflammatory mediators in patients with CHF were markedly elevated compared with healthy controls (P < or = .002). Methotrexate (n = 30) reduced plasma levels of tumor necrosis factor alpha (-15.6%, P < .05), interleukin 6 (-21.8%, P < .01), monocyte chemoattractant protein-1 (-22.6%, P < .01), soluble intercellular adhesion molecule-1 (-19.2%, P < .05), and C-reactive protein (-27.2%, P < .01) compared with baseline. Furthermore, interleukin 10 (15.8 %, P < .05) and soluble IL-1 receptor antagonist (36.1%, P < .01) expression was increased, whereas improvements in NYHA classification, 6-minute walk test distance, and QOL were found compared with baseline. Monocyte chemoattractant protein-1 expression was lower and soluble IL-1 receptor antagonist expression higher in the MTX than placebo group (n = 32). Furthermore, the left ventricular ejection fraction, left ventricular end-diastolic dimension, and incidence of main adverse cardiac events between the 2 groups were similar. CONCLUSION: These results suggest that the addition of MTX to conventional therapy for CHF has significant anti-inflammatory effects and improves NYHA functional class, 6-minute walk test distance, and QOL. PMID- 16368294 TI - The impact of ejection fraction on outcomes after percutaneous coronary intervention in patients with congestive heart failure: an analysis of the National Heart, Lung, and Blood Institute Percutaneous Transluminal Coronary Angioplasty Registry and Dynamic Registry. AB - BACKGROUND: Patients with congestive heart failure (CHF) have higher rates of adverse outcomes after percutaneous coronary intervention (PCI). A comprehensive analysis of outcomes in patients with CHF in the current era has not been done. We studied the outcomes of patients with CHF who underwent PCI in the National Heart, Lung, and Blood Institute-sponsored Percutaneous Transluminal Coronary Angioplasty (PTCA) and Dynamic registries. METHODS: We evaluated demographic and angiographic characteristics and the clinical outcomes of patients with CHF in the Dynamic Registry and the PTCA Registry, excluding patients with acute myocardial infarction. In the Dynamic Registry, patients with CHF (n = 503) were compared with patients without CHF (n = 4194), and patients with CHF with a preserved ejection fraction (EF) (n = 134) were compared with patients with CHF who have a reduced EF (n = 199). The patients with CHF in the 1997 through 2001 Dynamic Registry (n = 236) were then similarly compared with patients with CHF in the earlier PTCA Registry (n = 117). RESULTS: In the Dynamic Registry, compared with patients without CHF, patients with CHF had a higher-risk clinical and angiographic profile, and a higher mortality rate both inhospital (2.6% vs 0.4%, P < or = .001) and at 1 year (13.1% vs 3.0%, P < .001). Patients with reduced EF had higher inhospital mortality rates and a trend toward higher mortality at 1 year. The patients with CHF in the Dynamic Registry compared with those in the PTCA Registry had a higher risk profile yet had significantly higher procedural success rates and improved clinical outcomes. CONCLUSIONS: Although CHF remains a strong predictor of adverse outcomes after PCI, significant improvement seen in the past decade is likely related to improved procedural techniques and improved medical therapy. PMID- 16368295 TI - The association between blood pressure and mortality in patients with heart failure. AB - BACKGROUND: The association between low blood pressure and prognosis in the general population has been controversial, with some reports suggesting an increased mortality for patients with the lowest blood pressures. Whereas many standard heart failure therapies decrease blood pressure, the relationship between mortality and blood pressure in patients with heart failure has not been previously evaluated. METHODS: We used the Digitalis Investigation Group trial database to evaluate retrospectively the relationship among systolic blood pressure (SBP), diastolic blood pressure (DBP), and survival among 5747 patients with New York Heart Association class II or III heart failure and left ventricular ejection fraction < or = 0.45. Cox proportional hazards models were used to identify covariates predictive of long-term mortality. RESULTS: The adjusted all-cause mortality rate during the entire study period for patients in the lowest SBP group (< 100 mm Hg) was 50% and was significantly higher than that of the reference group of patients with SBP of 130 to 139 mm Hg, which had a mortality rate of 32% (hazard ratio 1.65, 95% CI 1.25-2.17, P < .001). The relationship between SBP and mortality was significant (P < .001) and nonlinear (P = .009). The relationship between DBP and mortality was significant (P < .001), with the highest mortality seen in patients with DBP < 60 mm Hg. CONCLUSIONS: In patients with systolic dysfunction (left ventricular ejection fraction < or = 0.45) and New York Heart Association classes II and III symptoms, lower SBPs and DBPs were associated with greater mortality. PMID- 16368296 TI - Overview of acute decompensated heart failure in Argentina: lessons learned from 5 registries during the last decade. AB - BACKGROUND: The acute decompensated heart failure (ADHF) is not as well characterized as the chronic phase, particularly in Latin American countries. Thus, the aim of this overview was to describe the clinical profile, treatment, and inhospital course of ADHF during the last decade in Argentina. METHODS: Results obtained from 5 Argentinean prospective and multicenter registries, involving 2974 patients admitted for ADHF, were assessed. These registries were performed and published between 1992 and 2004. RESULTS: The mean age was 65 to 70 years, and nearly 40% were female. Coronary artery disease was the main etiology in nearly 30% of the patients. Between 1992 and 2004, the use of angiotensin converting enzyme inhibitors increased from 29.9% to 53.4% before admission and from 48.5% to 69.3% before discharge; the use of beta-blockers rose from 4.2% to 33.2% at admission and from 2.5% to 42.4% at predischarge (all P < .0001). Inhospital mortality rates in the first to the fifth registries were 12.1%, 4.6%, 10.5%, 8.9%, and 4.7% (P [trend] = .006). However, there were 98 (7.7%) deaths among 1272 patients before 2002, compared with 129 (7.6%) among 1702 since 2002 (P = .9). CONCLUSIONS: The clinical profile of this largest sample of ADHF reported from a Latin American country is different from that observed in clinical trials and comparable to registries worldwide. Although an improvement in the use of recommended drugs was observed in the last decade, the average mortality has not changed. These findings might have implications in the design of multinational clinical trials. PMID- 16368297 TI - An unexpected inverse relationship between HbA1c levels and mortality in patients with diabetes and advanced systolic heart failure. AB - BACKGROUND: In diabetes, poor glycemic control, as indexed by hemoglobin A1c (HbA1c), is associated with increased risk of cardiovascular events and new-onset heart failure (HF). However, in patients with diabetes and HF, the relationship between glucose control and survival has not been investigated. Our study aimed to evaluate the relationship between HbA1c levels and mortality in patients with diabetes and advanced systolic HF. METHODS: We studied a cohort of 123 patients with diabetes and advanced systolic HF referred to a single center with HbA1c values measured at presentation. The patients were grouped based on HbA1c: HbA1c < or = 7.0 (n = 49) and HbA1c > 7.0 (n = 74). RESULTS: The cohort was 70% men, ejection fraction of 25% +/- 7, 59% ischemic etiology, HbA1c 7.9 +/- 1.8, and diabetes duration of 8.6 +/- 9.0 years. The HbA1c groups were similar in age; sex; New York Heart Association class; body mass index; diabetes duration; and insulin, metformin, and glitazone use. HbA1c > 7.0 was associated with higher ejection fraction, increased beta-blocker, and sulfonlyurea use. Patients with HbA1c < or = 7.0 had significantly increased all-cause mortality, compared with those with HbA1c > 7.0 (35% vs 20%, hazard ratio 2.6, 95% CI 1.3-5.2, P < .01). In multivariate analysis, HbA1c < or = 7.0 remained associated with increased mortality (hazard ratio 2.3, 95% CI 1.0-5.2). CONCLUSIONS: Paradoxically, elevated HbA1c levels were associated with improved survival in this cohort of patients with diabetes and advanced HF. Further investigation is necessary to determine the nature of this relationship and optimal HbA1c in patients with diabetes and HF. PMID- 16368298 TI - Valsartan inhibits platelet activity at different doses in mild to moderate hypertensives: Valsartan Inhibits Platelets (VIP) trial. AB - BACKGROUND: Previous in vitro studies have suggested that valsartan produces significant inhibition of human platelets, probably targeting angiotensin I platelet receptors. To test whether valsartan inhibits platelet activity in mild to moderate hypertensives we conducted the randomized Valsartan Inhibits Platelets (VIP) trial. METHODS AND RESULTS: Seventy-five patients with mild to moderate hypertension were randomized to valsartan 80 (n = 25), valsartan 160 (n = 29), or valsartan 320 mg/d (n = 21) for 9 weeks. Platelet function was assessed at baseline, week 5, and week 9 by aggregometry, flow cytometry, and cartridge based analyzer. Independently of dose and duration, valsartan provided early sustained significant inhibition of adenosine diphosphate-induced platelet aggregation, decreased shear-induced activation measured with PFA-100 analyzer, and diminished expression of GP IIb/IIIa activity measured by PAC-1 antibody, GPIb (CD42b), vitronectin receptor (CD51/61), P-selectin (CD62p), lysosome associated membrane protein (CD107a), and CD40-ligand (CD154). The antiplatelet properties of valsartan were more profound in patients with diabetes (n = 28) when compared with the nondiabetic group (n = 47). In subgroup analyses of patients with diabetes there appeared to be stronger inhibition of the platelet receptors, a significant decrease of adenosine diphosphate- and collagen-induced platelet aggregation, and more profound inhibition of GP IIb/IIIa activity. CONCLUSIONS: In the randomized VIP trial, valsartan produced sustained inhibition of platelet aggregation and major platelet receptors. The antiplatelet properties of valsartan were not dose or time dependent. In subgroup analyses patients with diabetes with mild to moderate hypertension tended to have greater platelet inhibition, a finding which, if confirmed in future studies suggests possible additional advantages for using valsartan in this high-risk population. PMID- 16368299 TI - Natural antioxidants from tomato extract reduce blood pressure in patients with grade-1 hypertension: a double-blind, placebo-controlled pilot study. AB - BACKGROUND: Treatment of hypertension (HT) can reduce the risk for cardiovascular diseases. Tomato extract contains carotenoids such as lycopene, beta carotene, and vitamin E, which are known as effective antioxidants, to inactivate free radicals, and to slow the progression of atherosclerosis. The purpose of our study was to evaluate the effect of tomato extract on systolic and diastolic blood pressure in grade-1 HT, on serum lipoproteins, plasma homocysteine, and oxidative stress markers. METHODS: This study is a single-blind, placebo controlled trial. Thirty-one subject with grade-1 HT, without concomitant diseases, who required no antihypertensive or lipid-lowering drug therapy, who were recruited from primary care clinics, completed the trial. Subjects entered a 4-week placebo period, then an 8-week treatment period with tomato extract, 250 mg Lyc-O-Mato, and a 4-week control period with placebo. RESULTS: Systolic blood pressure decreased from 144 (SE +/- 1.1) to 134 mm Hg (SE +/- 2, P < .001), and diastolic blood pressure decreased from 87.4 (SE +/- 1.2) to 83.4 mm Hg (SE +/- 1.2, P < .05). No changes in blood pressure were demonstrated during placebo periods. Thiobarbituric acid-reactive substances, a lipid peroxidation products marker, decreased from 4.58 (SE +/- 0.27) to 3.81 nmol/mg (SE +/- 0.32, P < .05). No significant changes were found in lipid parameters. CONCLUSIONS: A short-term treatment with antioxidant-rich tomato extract can reduce blood pressure in patients with grade-1 HT, naive to drug therapy. The continuous effect of this treatment and the long-term beneficial effect on cardiovascular risk factors still need to be demonstrated. PMID- 16368300 TI - Identification of the ischemic etiology of heart failure by cardiovascular magnetic resonance imaging: diagnostic accuracy of late gadolinium enhancement. AB - BACKGROUND: A large proportion of patients with heart failure (HF) have a large and poorly contracting left ventricle. The noninvasive recognition of the ischemic etiology of such patients is difficult, and for this purpose, usually patients undergo coronary angiography. It has been shown that cardiovascular magnetic resonance (CMR) imaging can detect myocardial scarring by evaluating late gadolinium enhancement (LGE). The diagnostic accuracy of such method in differentiating the etiology of HF has not been previously tested in an unselected HF ambulatory population. METHODS: We studied 60 ambulatory patients consecutively enrolled from a specialized HF clinic. We included HF patients who were found to have increased left ventricular (LV) dimensions and reduced function. CMR was performed in these patients by operators who were unaware of patients' history and clinical conditions. LV dimensions and global and regional function, as well as the pattern of LGE, were obtained in each subject. Coronary angiography was subsequently performed in all the patients. The diagnostic accuracy of clinical history and electrocardiographic patterns, as well as regional wall motion abnormalities, wall thinning, and LGE, in differentiating coronary artery disease (CAD) from non-CAD patients were evaluated. RESULTS: The majority of CAD patients (98%) showed LV contrast hyperenhancement with respect to non-CAD HF subjects (16%). The detection of LGE by CMR had a sensitivity of 98% and a specificity of 84% and an overall accuracy of 93% in detecting CAD etiology among HF patients. CONCLUSIONS: LGE is able to accurately differentiate CAD from non-CAD etiology of HF and may represent a clinically useful noninvasive tool for this purpose. As it provides relevant functional information as well as insight into the etiology, CMR may be included among the most important diagnostic tools in the workup of patients with HF. PMID- 16368301 TI - Regional diastolic dysfunction in individuals with left ventricular hypertrophy measured by tagged magnetic resonance imaging--the Multi-Ethnic Study of Atherosclerosis (MESA). AB - BACKGROUND: Impairment of global diastolic function is considered to be the mechanism of congestive heart failure in individuals with preserved systolic left ventricular (LV) function. Left ventricular hypertrophy (LVH) is known to be a risk factor for congestive heart failure with preserved systolic function, and this process may begin as a regional process. We investigated whether regional LV diastolic function measured by magnetic resonance tagging is altered in asymptomatic participants of the MESA with LVH and preserved systolic LV function. METHODS: Regional systolic and diastolic strain rates were calculated from strain data in 218 participants of the MESA study. Circumferential strain was calculated from the midwall layer of the septum, anterior, lateral, and inferior walls at mid-LV level. Global LV function measures were studied by magnetic resonance imaging in 4291 MESA participants. Left ventricular hypertrophy for men and women was defined from the MESA population using previously established Framingham criteria. RESULTS: Global systolic function was slightly less in the LVH (ejection fraction = 0.66 +/- 0.10) versus the non-LVH group (ejection fraction = 0.69 +/- 0.07, P < .001). Stepwise regression analyses showed a direct relationship between regional diastolic dysfunction and increasing LV mass. Regional systolic strain and strain rate measures from participants with LVH were not significantly different from those without LVH. However, regional diastolic strain rate was significantly reduced in participants with LVH (1.5 +/- 1.1 s(-1)) compared with the non-LVH group (2.2 +/- 1.1 s(-1), P < .001) regardless of age or sex. CONCLUSIONS: Left ventricular hypertrophy is associated with regional diastolic dysfunction in individuals without evidence of clinical cardiovascular disease and preserved systolic function. Magnetic resonance imaging tagging provides detailed quantification of regional diastolic function noninvasively. PMID- 16368302 TI - Time course of left ventricular volumes in severe congestive heart failure patients treated by optimized AV sequential left ventricular pacing alone--a 3 dimensional echocardiographic study. AB - BACKGROUND: This study evaluates the acute and chronic resynchronizing effects of AV sequential left ventricular (LV) pacing on LV function in patients with impaired cardiac function and conduction disorders by 3-dimensional transesophageal echocardiography. METHODS AND RESULTS: Twenty-nine patients with congestive heart failure, with LV ejection fraction (LVEF) < or = 30%, QRS duration > or = 120 milliseconds, and New York Heart Association Class II to IV, were implanted with a cardiac resynchronization device using an LV lead only, according to the invasively determined hemodynamic optimal pacing site and AV delay. Patients underwent 3-dimensional transesophageal echocardiography before randomization to treatment (baseline) and at 12-month follow-up (resynchronization--12 months). Three-dimensional volumes were acquired on resynchronization and during intermittent switch-off at intrinsic depolarization. The values of stroke volume were 43.2 +/- 13.3 (intrinsic-baseline), 51.7 +/- 17.4 (intrinsic--12 months), 57.2 +/- 15.6 (resynchronization-baseline), and 64.6 +/- 18.9 (resynchronization--12 months). Analysis of variance demonstrated a significant effect of resynchronization at different periods (P < .001) and a significant time effect (P < .05) for stroke volume. Similar results were observed with ejection fraction (LVEF). No effect was observed with LV end diastolic volume, whereas a therapy effect with no time effect was observed with LV end-systolic volume. CONCLUSIONS: A significant acute increase of LV stroke volume and LVEF was found by resynchronization by LV pacing alone. A continuous improvement of LV stroke volume and LVEF occurred with time of follow-up (reverse remodeling). The initial therapeutic effect persisted during 12-month follow-up independently of time of follow-up and QRS width. No significant decrease of LV end-diastolic size during chronic resynchronization was detected in contrast to previous studies with resynchronization by biventricular pacing. PMID- 16368303 TI - The additional prognostic value of coronary flow reserve on left anterior descending artery in patients with negative stress echo by wall motion criteria. A Transthoracic Vasodilator Stress Echocardiography Study. AB - BACKGROUND: Vasodilator stress echocardiography allows imaging of left anterior descending (LAD) coronary flow reserve (CFR), which may provide additional prognostic information over regional wall motion. AIM: To assess the prognostic value of CFR in patients with known or suspected coronary artery disease (CAD) and negative stress echo. METHODS: We studied 329 consecutive patients (193 men, age 61 +/- 13 years) with known (n = 101) or suspected (n = 228) CAD and negative stress echo by standard wall motion criteria. All patients underwent dipyridamole (up to 0.84 mg/kg in 10 minutes) stress echo with CFR evaluation of LAD by Doppler. RESULTS: During follow-up (28 +/- 10 months), 22 events occurred: 1 cardiac death, 6 nonfatal myocardial infarctions, 5 unstable anginas, and 10 late (> 6 months) coronary revascularizations. Moreover, 9 patients underwent early (< 6 months) revascularization and were censored. Using a receiver operating characteristic analysis, CFR < or = 1.92 was the best predictor of future events (area under the curve = 0.80, sensitivity = 77%, specificity = 85%) and was taken as criterion for reduced CFR accordingly. Sixty-three (19%) patients had reduced and 266 (81%) had normal CFR on LAD. The 36-month event-free survival was higher in patients with normal and lower in patients with reduced CFR (98% vs 64%, P < .0001). At Cox analysis, CFR of LAD (hazard ratio [HR] 16.52, 95% CI 5.76-47.40, P < .0001), left ventricular mass index (HR 1.03 per unit increment, 95% CI 1.00 1.05, P = .01), and smoking habit (HR 3.00, 95% CI 1.24-7.23, P = .01) were independent prognostic indicators. Using an interactive stepwise procedure, CFR on LAD provided additional prognostic information to clinical, resting echo, and angiographic findings. CONCLUSIONS: In patients with known or suspected CAD and negative stress echocardiography by wall motion criteria, CFR provides independent information for prognostic stratification, and a reduced CFR is associated with a less benign long-term outcome. PMID- 16368304 TI - Real-time myocardial perfusion imaging for pharmacologic stress testing: added value to single photon emission computed tomography. AB - BACKGROUND: Little is known about the incremental value of real-time myocardial contrast echocardiography (MCE) as an adjunct to pharmacologic stress testing. This study was performed to evaluate the diagnostic value of MCE to detect abnormal myocardial perfusion by technetium Tc 99m sestamibi-single photon emission computed tomography (SPECT) and anatomically significant coronary artery disease (CAD) by angiography. METHODS: Myocardial contrast echocardiography was performed at rest and during vasodilator stress in consecutive patients (N = 120) undergoing SPECT imaging for known or suspected CAD. Myocardial opacification, wall motion, and tracer uptake were visually analyzed in 12 myocardial segments by 2 pairs of blinded observers. Concordance between the 2 methods was assessed using the kappa statistic. RESULTS: Of 1356 segments, 1025 (76%) were interpretable by MCE, wall motion, and SPECT. Sensitivity of wall motion was 75%, specificity 83%, and accuracy 81% for detecting abnormal myocardial perfusion by SPECT (kappa = 0.53). Myocardial contrast echocardiography and wall motion together yielded significantly higher sensitivity (85% vs 74%, P < .05), specificity of 83%, and accuracy of 85% (kappa = 0.64) for the detection of abnormal myocardial perfusion. In 89 patients who underwent coronary angiography, MCE and wall motion together yielded higher sensitivity (83% vs 64%, P < .05) and accuracy (77% vs 68%, P < .05) but similar specificity (72%) compared with SPECT for the detection of high-grade, stenotic (> or = 75%) coronary lesions. CONCLUSION: Assessment of myocardial perfusion adds value to conventional stress echocardiography by increasing its sensitivity for the detection of functionally abnormal myocardial perfusion. Myocardial contrast echocardiography and wall motion together provide higher sensitivity and accuracy for detection of CAD compared with SPECT. PMID- 16368305 TI - Effect of atorvastatin on circulating proinflammatory T-lymphocyte subsets and soluble CD40 ligand in patients with stable coronary artery disease--a randomized, placebo-controlled study. AB - BACKGROUND: Coronary atherosclerosis includes an activation of circulating T lymphocytes. Statins exert anti-inflammatory effects beyond lipid lowering. Whether these properties influence systemic T lymphocytes is unclear. METHODS: To investigate the effect of atorvastatin on circulating T-lymphocyte subsets producing proinflammatory and anti-inflammatory cytokines (interferon gamma [IFN gamma(+)], interleukin 2 [IL-2(+)], IL-4(+), and IL-10(+)) and on the T-cell activating soluble CD40 ligand (sCD40L), 30 hypercholesterolemic patients with angiographically documented stable coronary artery disease (CAD) were randomized to placebo or atorvastatin (20 mg/d) for 3 months. Eight healthy volunteers served as controls. Levels of peripheral cytokine-producing CD4+ and CD8+ T cells and their CD28- subsets were determined by FACS. Serum soluble CD40L was measured with ELISA. RESULTS: IL-2(+) T lymphocytes and sCD40L levels were higher in patients with CAD compared with controls, whereas IFN-gamma(+) and anti inflammatory IL-4(+) and IL-10(+) T lymphocytes were similar. Levels of IL-2(+), IFN-gamma(+), IL-4(+), and IL-10(+) T-cell subsets as well as CD28- T lymphocytes were neither changed by atorvastatin nor by placebo, whereas sCD40L was lowered only in atorvastatin-treated patients (P < .01). CONCLUSION: Circulating IL-2(+) T lymphocytes are increased in patients with stable CAD reflecting an activation of the global immune system, but are not influenced by atorvastatin therapy. The elevated levels of platelet-derived T-lymphocyte-stimulating sCD40L are decreased by atorvastatin probably reflecting an atheroprotective effect. Hence, sCD40L may be an additional biomarker to be considered when evaluating the treatment effects of statins in patients with stable CAD. PMID- 16368306 TI - Impact of body mass index on outcomes of enhanced external counterpulsation therapy. AB - OBJECTIVES: We evaluated the association of baseline body mass index (BMI) on the outcomes of enhanced external counterpulsation (EECP) therapy for chronic stable angina. BACKGROUND: We are in the midst of a pandemic of obesity, which is complicating the care of patients with coronary artery disease (CAD). METHODS: We examined 2730 patients enrolled from 2002 to 2004 in the IEPR-2. Baseline and outcome variables were stratified by the entry BMI in kilograms per meter squared. RESULTS: Obesity (BMI > 30 kg/m2) was common (40.6%) among patients with severe CAD referred for EECP. Within the total cohort, 2.6% was underweight (BMI < or = 20 kg/m2) and 4.5% was morbidly obese (BMI > 40 kg/m2). Prevalence of diabetes, hypertension, dyslipidemia, and heart failure (HF) was higher in obese patients. However, the rates of baseline angina and prior revascularization were similar among the groups. The peak diastolic augmentation ratio was similar between groups during the first (0.7 +/- 0.4 for lowest and highest BMI) and last hours of treatment (0.9 +/- 0.5 and 0.8 +/- 0.5). The cumulative hours of treatment, the change in angina class, and the Duke Activity Status Index were similar for all BMI groups. There was a greater reduction in weekly anginal episodes from baseline across ascending levels of BMI (-6.3 +/- 13.6 to -9.7 +/- 15.8, P = .03). The rates of discontinuation for clinical events were highest (14.3%) with skin breakdown being the most frequent cause (10.1%) in the underweight. The rates of clinical events including myocardial infarction, HF, and death trended higher across ascending levels of BMI (P = .52). Multivariate analysis found that older age, history of stroke, history of HF, and diabetes, but not BMI, were predictors of clinical events. CONCLUSIONS: More than 40% of patients with severe CAD referred for EECP were obese. Underweight patients had higher rates of discontinuation of treatment mainly because of skin breakdown. Symptomatic benefit of EECP was similar among all BMI groups. However, despite symptomatic improvement, there was a nonsignificant trend for higher rates of myocardial infarction, HF, and death as BMI increased. PMID- 16368307 TI - N-acetylcysteine and contrast-induced nephropathy: a meta-analysis of 13 randomized trials. AB - BACKGROUND: Contrast-induced nephropathy (CIN) following coronary angiography increases morbidity and mortality. Randomized trials of small sample size have evaluated whether N-acetylcysteine (NAC) prevents CIN in patients with renal dysfunction. METHODS: To conduct a meta-analysis of the randomized trials the following databases were searched: MEDLINE (1966-2003), Cochrane Controlled Trials Register, ACP Journal Club online, published abstracts presented at the major cardiology and nephrology meetings, references from reviews. Two authors independently evaluated all relevant randomized trials. Eligibility criteria were (1) randomized placebo controlled trials of NAC, (2) patients with impaired renal function (creatinine >1.2 mg/dL) undergoing coronary angiography, (3) patients receiving intravenous fluids and low-osmolarity nonionic contrast media, (4) the primary outcome was CIN (increases in creatinine of either at least 0.5 mg/dL or 25% from baseline to 48 hours). Of 589 trials reviewed 3 disagreements were easily resolved by mutual discussion and 13 were selected. Data extraction included patient characteristics, intravenous fluid regimen, type and dose of contrast media, dosing regimen, creatinine at baseline and 48 hours and CIN requiring dialysis. RESULTS: Four of the 13 trials reported statistically significant results. In meta-analysis of the 13 trials, which included 1892 patients, the RR was 0.68 (95%CI, 0.46-1.01). The addition of the trial of patients undergoing computerized tomography, which had formulated the hypothesis, yielded a statistically significant reduction (RR 0.64 [95%CI 0.42-0.96]) as did an earlier meta-analysis of 7 trials. CONCLUSIONS: Our meta-analysis of the most currently available randomized data concerning NAC before coronary angiography to prevent CIN in patients with impaired renal function is neither conclusive nor provides proof beyond a reasonable doubt to influence clinical practice and public policy. The intervention has minimal toxicity but the width of the 95% CI remains compatible with a range from a large benefit to none at all. In addition, the trials used change in creatinine as the measure of outcome. Further randomized trials of large sample size and with clinical outcomes will add importantly relevant information to the totality of evidence and allow the most rational clinical decisions for individual patients as well as policy decisions for the health of the general public. PMID- 16368308 TI - Impact of renal insufficiency on outcome after contemporary percutaneous coronary intervention. AB - BACKGROUND: End-stage renal failure is associated with poor outcomes, including increased mortality, after percutaneous coronary intervention (PCI). The effect of milder degrees of renal insufficiency (RI) is less clear, especially with routine stenting and glycoprotein IIb/IIIa inhibitor therapy, which may be of particular benefit in patients with RI. METHODS: Clinical, angiographic, procedural, and outcome variables of 7769 consecutive patients who underwent PCI between April 2000 and July 2004 were entered into a prospective database. Inhospital mortality and morbidity were calculated according to baseline creatinine clearance. Simple and multiple logistic regression analyses were performed to determine independent predictors of mortality. RESULTS: Baseline creatinine clearance was available in 6840 patients. It was normal (> 80 mL/min) in 3474; 1670 had mild RI (61-80 mL/min), 1111 moderate RI (41-60 mL/min), and 585 severe RI (< or = 40 mL/min). Major adverse cardiac events (MACE) (death/myocardial infarction/revascularization) increased substantially with worsening renal function (2.4% vs 3.0% vs 4.8% vs 9.7%, P < .0001), as did mortality (0.3% vs 0.7% vs 1.5% vs 6.0%, P < .0001). Multiple logistic regression analysis identified moderate RI and severe RI as independent predictors of mortality (odds ratio [OR] 3.9, P < .001; OR 12.7, P < .0001, respectively) and morbidity (MACE) (OR 1.5, P < .05; OR 2.5, P < .0001, respectively). Mild RI trended to increase the risk of mortality but did not reach statistical significance as an independent predictor of inhospital death on multiple regression analysis (OR 2.1, P = .1) and did not increase the risk of MACE (OR 1.1, P = .6). CONCLUSIONS: Despite routine stenting and glycoprotein IIb/IIIa inhibitor therapy, RI remains an independent predictor of increased morbidity, and particularly mortality, after PCI. However, the adverse effect of truly mild RI on outcome is limited. PMID- 16368309 TI - Side branch occlusion after coronary stent implantation in patients presenting with ST-elevation myocardial infarction: clinical impact and angiographic predictors. AB - BACKGROUND: The aim of this study was to assess the incidence and clinical outcome of the occlusion of major (> 1 mm) side branches following coronary stenting in patients undergoing percutaneous coronary intervention for acute ST elevation myocardial infarction (STEMI). METHODS: Among 276 consecutive patients presenting with STEMI, we found 80 patients (29%) with 101 stent-covered side branches. Clinical data and quantitative angiographic analysis were evaluated. Angiographic follow-up was available in 56 (70%) patients, and clinical follow-up could be completed in all patients. RESULTS: Acute side branch occlusion after stent implantation (SBO) was observed in 10 (12.5%) patients involving 11 (10.9%) side branches. Predictors for SBO were: (1) reference side branch diameter at baseline < or = 1.4 mm; (2) ostial side branch stenosis > 50%; and (3) minimal side branch diameter at baseline < or = 0.6 mm. During hospitalization, in the SBO group, 2 patients died in cardiogenic shock and 1 underwent bypass surgery; no events were causally related to SBO. During long-term follow-up, 1 patient with SBO developed repeat MI as opposed to 7 patients in the non-SBO group who developed major adverse cardiac events (1 death, 6 repeat revascularizations). CONCLUSIONS: The presence of a side branch originating from the target lesion in patients undergoing coronary stenting for acute STEMI is a frequent observation (29%) and is associated with a low incidence of side branch occlusion. Major predictors for SBO are the side branch size and the presence of an ostial side branch stenosis. PMID- 16368310 TI - Incidence and predictors of late recurrence after beta-radiation therapy with a 188Re-MAG3-filled balloon for diffuse in-stent restenosis. AB - BACKGROUND: The long-term fate of patent irradiated segments at 6 months after beta-radiation therapy has not been sufficiently evaluated. METHODS: Two-year follow-up angiography was performed in 52 patients with patent irradiated segments at 6 months after beta-radiation with a rhenium 188 mercaptoacetyltriglycine-filled balloon for diffuse in-stent restenosis. We evaluated late recurrence (LR) and its predictors after beta-radiation. RESULTS: Late recurrence at 2 years after radiation was observed in 10 (19.2%) of 52 patients. The minimal lumen diameter (MLD) progressively decreased, from 2.67 +/- 0.44 mm at postprocedure to 2.42 +/- 0.53 mm at 6 months to 2.09 +/- 0.75 mm at 2 years (P = .001). In the 42 patients without LR, the MLD decreased from postprocedure (2.74 +/- 0.43 mm) to 6 months (2.44 +/- 0.54 mm; P = .006), but did not change between 6 months and 2 years (2.35 +/- 0.49 mm, P = .13). In the LR group, the MLD was unchanged from postprocedure (2.33 +/- 0.29 mm) to 6 months (2.30 +/- 0.43 mm; P = .81), but decreased significantly between 6 months and 2 years (1.02 +/- 0.75 mm, P = .001). Multivariate analysis identified postprocedural MLD as an independent predictor of LR (odds ratio 0.025, 95% CI 0.007-0.94, P = .04). Late target lesion revascularization was performed in 6 patients (11.5%) between 6 months and 2 years after radiation. CONCLUSION: Although LR after radiation was observed in some patients, irradiated segments remained stable for up to 2 years in most patients. Smaller postprocedural MLD, followed by delayed late loss between 6 months and 2 years, was associated with LR. PMID- 16368311 TI - Impact of race and ethnicity on inhospital outcomes after percutaneous coronary intervention (report from the 2000-2001 New York State Angioplasty Registry). AB - BACKGROUND: Studies regarding the impact of race and ethnicity on outcomes after percutaneous coronary intervention (PCI) in the modern era are limited. METHODS: Using the 2000 and 2001 New York State PCI Databases, we compared baseline clinical, demographic, and angiographic characteristics and subsequent inhospital events among 76,928 patients of black, Hispanic, and white racial/ethnic backgrounds. We sought to determine the influence of race and ethnicity, if any, on post-PCI outcomes. RESULTS: Blacks and Hispanics were younger and more likely to be hypertensive, diabetic, obese, in congestive heart failure, and have chronic renal insufficiency. Whites were more likely to be men, have multivessel disease, and receive a stent. There was no significant difference in unadjusted post-PCI inhospital mortality (0.7% for all groups) or major adverse cardiac event (defined as death, emergent coronary bypass, or stroke) among all 3 racial groups. After correcting for clinical and demographic variables, race/ethnicity was not a significant predictor of death or major adverse cardiac event. CONCLUSION: Minority patients of black and Hispanic decent have a significantly higher incidence of traditional cardiovascular risk factors and present for angioplasty at a younger age compared with whites. However, there is no significant difference in outcomes after angioplasty among these racial/ethnic groups. PMID- 16368312 TI - Impact of body mass index on outcomes after primary angioplasty in acute myocardial infarction. AB - BACKGROUND: The prognostic importance of obesity after primary percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI) is unknown. We therefore sought to investigate the impact of body mass index (BMI) in patients with AMI undergoing primary PCI. METHODS: In the CADILLAC trial, 2082 patients of any age with AMI within 12 hours onset undergoing primary PCI were randomized to balloon angioplasty versus stenting, each +/-abciximab. Outcomes were stratified by baseline BMI. RESULTS: Baseline BMI was measured in 2035 (98%) randomized patients; 552 (27%) patients have normal weight (BMI < 25 kg/m2), 915 (45%) were overweight (> or = 25 to < 30 kg/m2), and 568 (28%) were obese (> or = 30 kg/m2). Compared with normal-weight patients, obese patients were younger and more frequently had diabetes, hyperlipidemia, hypertension, non-anterior myocardial infarction, and higher creatinine clearance. Obese patients were less likely to develop thrombocytopenia (1.8% vs 4.2%), moderate hemorrhagic complications (1.4% vs 3.3%), or required blood product transfusions (3.2% vs 6.3%) (all P < or = .04). Obese compared with normal-weight patients had lower inhospital mortality (0.9% vs 2.7%, P = .03) at 30 days (1.1% vs 3.8%, P = .02) and 1 year (1.8% vs 7.5%, P < .0001). Independent predictors of 30-day and 1-year mortality included lower ejection fraction, advanced age, 3-vessel disease, anterior AMI, and lower creatinine clearance, but not BMI. CONCLUSIONS: Obese patients with AMI have an improved prognosis after primary PCI compared with normal-weight patients, a finding attributable to AMI onset at younger age, with better renal function and less anterior infarction. PMID- 16368313 TI - A randomized pilot study of dalteparin versus unfractionated heparin during percutaneous coronary interventions. AB - BACKGROUND: Direct comparison of low-molecular-weight heparin, dalteparin, with unfractionated heparin (UFH) during percutaneous coronary intervention (PCI) is limited. This study examined the relative effects of dalteparin and UFH on coagulation and angiographic and clinical indices during PCI. METHODS: This was a double-blind randomized study, stratified by planned glycoprotein IIb/IIIa inhibitor use. Both UFH and dalteparin were administered as an intra-arterial bolus immediately before PCI. RESULTS: All randomized patients received the assigned study drug and underwent PCI. Mean activated clotting time levels were 344 seconds for UFH and 234 seconds for dalteparin (P < .0001). Anti-factor Xa levels were higher for dalteparin at 30 minutes (UFH 1.3 IU/mL vs dalteparin 1.7 IU/mL, P = .005)) and at 4 hours (UFH 0.27 IU/mL vs dalteparin 0.69 IU/mL, P < .0001). Angiographic success was > 90% in both groups, and angiographic complications were similar (UFH 2.5% vs dalteparin 3.8%). The composite of death, myocardial infarction, target vessel revascularization, or bailout glycoprotein IIb/IIIa at hospital discharge was 13.7% in the UFH group and 13.1% in the dalteparin group (P = not significant). There were 2 major bleedings requiring transfusion, both occurring in the UFH group. CONCLUSIONS: This study suggests that a single intra-arterial bolus of low-molecular-weight heparin without monitoring is feasible and warrants further investigation as an alternative to UFH during PCI. PMID- 16368314 TI - Measuring the efficacy of antihypertensive therapy by ambulatory blood pressure monitoring in the primary care setting. AB - BACKGROUND: Traditional clinical trials in hypertension measure the efficacy of antihypertensive drugs but may not fully assess their effectiveness in clinical practice. Community-based trials can provide this information but are limited because usually they are of open-label design and potentially subject to observer bias. Therefore, we used ambulatory blood pressure monitoring (ABPM), an automated and objective measure of blood pressure (BP) to overcome these shortcomings in a large community-based trial. METHODS: Patients with hypertension, either untreated or currently on treatment, were started on, or switched to, the angiotensin receptor blocker telmisartan 40 mg daily; after 2 weeks, if office BP remained > or = 140/85 mm Hg, the dose was increased to 80 mg, and if necessary, hydrochlorothiazide 12.5 mg was added after a further 4 weeks and continued for the final 4-week period. Baseline and treatment ABPM measurements were completed in 940 previously untreated patients and 675 previously treated patients. RESULTS: The average reduction of the entire cohort was -10.7/-6.5 mm Hg (P < .0001; mean 24-hour BPs were reduced by 12/8 and 8/5 mm Hg in the untreated and previously treated patients, respectively). In contrast, the office BPs fell by an average of 23/12 and 17/10 mm Hg in previously untreated and treated patients. In 401 patients whose baseline 24-hour BP was > or = 130/85 mm Hg, the mean decrease in 24-hour BP was 16.8/11.4 mm Hg. Based on ABPM criteria, the BP was fully controlled (< 130/85 mm Hg) in 70% of patients, and based on office measurement criteria (< 140/90 mm Hg), in 79%. CONCLUSIONS: Ambulatory BP monitoring demonstrated excellent control rates by telmisartan monotherapy or in combination with hydrochlorothiazide. Observer and measurement bias was substantial based on the changes from baseline by clinical measurements in contrast to ambulatory BP recordings. The successful use of this procedure in primary care research will create further opportunities to define the effectiveness of treatment in the environment in which it is customarily prescribed. PMID- 16368315 TI - Long-term adherence with cardiovascular drug regimens. AB - BACKGROUND: An increasing number of medications are prescribed for patients with coronary artery disease, but poor adherence may limit realization of their benefits. OBJECTIVE: To characterize adherence to evidence-based cardiovascular medications prescribed at hospital discharge at 1 year. METHODS: We studied 1326 patients with coronary artery disease undergoing cardiac catheterization between 1998 and 2001. We examined adherence to angiotensin-converting enzyme (ACE) inhibitors, aspirin, beta-blockers (BBs), and statins by comparing baseline prescription at hospital discharge to self-reported medical regimen at 12 months. Patients who reported use of each cardiac medication at 1 year were considered adherent. Clinical and demographic predictors of nonadherence are described. RESULTS: The population had a mean age of 65.7 +/- 10.5 years, and 36% were women. At discharge, aspirin was prescribed in 95%, BBs in 86%, ACE inhibitors in 65%, and statins in 55%. The proportion of patients who discontinued medications was lowest for aspirin (18%) and BBs (22%) and highest for ACE inhibitors/angiotensin receptor blockers (28%) and statins (28%). Only 54% were adherent to all of their initial medications. Patients who discontinued medications were more likely to be older, women, unmarried, and less educated. Multivariable predictors of better adherence were higher mental health, education level, marital status, and no antidepressant use. A higher number of prescribed medications were associated with lower adherence to the recommended regimen. Insurance coverage and physical function did not correlate with adherence. CONCLUSIONS: Patients frequently stop medications within 1 year of prescription. Adherence is influenced by marital status, mental health, education, and total number of medications prescribed. Physicians need to be aware of patient factors which influence adherence to facilitate higher use of evidence-based medications. PMID- 16368316 TI - Prevention of implantable defibrillator shocks by cognitive behavioral therapy: a pilot trial. AB - BACKGROUND: Although psychological stress is known to favor ventricular arrhythmic events, there is no evidence that stress management intervention decreases ventricular electrical instability in implantable cardioverter defibrillator (ICD) patients. The aim of the study was to determine whether cognitive behavioral therapy (CBT) results in a decrease of arrhythmic events requiring ICD intervention through an improvement in sympathovagal balance. METHODS: Of 253 consecutive ICD patients (age 59 +/- 10 years, 64 men), 70 were randomly assigned to CBT (n = 35) or conventional medical care (n = 35). Measures of heart rate variability, psychological well-being, and quality of life were assessed at baseline, 3 months, and 1 year. The primary outcome was appropriate ICD shock. RESULTS: Although, it was not statistically different, the number of patients requiring shocks was less in the CBT group than in the conventional treatment group. At 3 months, among patients without antiarrhythmic drugs, none of the subjects in the CBT group had experienced arrhythmic events requiring ICD intervention, as compared with 4 in the control group (P < .05). At 12 months, there was no difference in the number of arrhythmic events requiring therapy between the CBT group versus the control group. Among heart rate variability indexes, daytime pNN 50 and nocturnal SDNN improved significantly in the CBT group, as compared with the control group. CONCLUSIONS: By decreasing anxiety and possibly improving sympathovagal balance, cognitive behavior therapy may decrease the propensity for ventricular arrhythmias in ICD patients. However, these effects appear to be limited over time. PMID- 16368317 TI - Autologous bone marrow mononuclear cell transplantation in patients undergoing coronary artery bypass grafting. AB - BACKGROUND: Recent studies have shown that autologous bone marrow mononuclear cell (aBM-MNC) transplantation can be effectively performed in human beings either by the coronary route or by endoventricular injections. However, scanty data are available for patients undergoing coronary artery bypass grafting (CABG). Accordingly, the aim of this study was to evaluate the feasibility and safety of aBM-MNC transplantation in patients with recent myocardial infarction undergoing CABG. METHODS AND RESULTS: The study population included 36 consecutive patients with recent myocardial infarction (< 6 months) undergoing CABG. Eighteen patients (17 men, mean age 64 years) underwent CABG plus aBM-MNC transplantation, whereas 18 subjects undergoing conventional CABG (17 men, mean age 67 years) served as control subjects. Cell transplantation was performed by direct injections in the border zone of the recently infarcted area. An average number of 292 +/- 232 x 10(6) aBM-MNCs was injected in each patient. When compared with control subjects, transplanted patients showed higher values of troponin I peak after CABG (median values of 1.65 ng/mL vs 0.64 ng/mL, P < .001). No major transplant-related adverse event could be detected. During follow-up, transplanted patients had an improvement in left ventricular ejection fraction (from 0.46 to 0.51, P < .05) and wall motion score index (from 1.71 to 1.42, P < .01). The incidence of arrhythmias immediately after CABG and during follow-up was similar in the 2 groups. CONCLUSIONS: Our data support the idea that direct injection of aBM-MNCs in the myocardium during CABG is feasible and safe. Larger studies are needed to assess the efficacy of such an approach in patients undergoing CABG. PMID- 16368318 TI - Prospective randomized study comparing coronary artery bypass grafting with the new mini-extracorporeal circulation Jostra System or with a standard cardiopulmonary bypass. AB - BACKGROUND: To assess the potential benefits of a new concept of cardiopulmonary bypass (CPB), the mini-extracorporeal circulation (MECC) Jostra System, we conducted a prospective randomized study among patients who underwent coronary artery bypass grafting (CABG) with a MECC Jostra System or with a standard CPB. METHODS: In a prospective randomized study, 400 patients underwent elective CABG using a standard CPB (200 patients) or a Jostra MECC System (200 patients). The patients were randomly assigned to have preoperative data similar for both groups. RESULTS: The operative mortality rate (< 30 days) was 1.5% for the MECC group and 2.5% for the CPB group (P = NS). Low-cardiac-output syndrome occurred in 0.66% and 4% for the CPB group (P < .001.). The inflammatory response was significantly reduced with the MECC. Likewise, C-reactive protein release was significantly higher in group B at 24 hours (69.6 +/- 38.5 vs 40.8 +/- 21.8 mg/L for the group A, P < .01) and at 48 hours (116.7 +/- 47.0 vs 65.4 +/- 39.5 mg/L, P < .05) postoperatively. In the CPB group, there was a significantly higher decrease of hematocrit and hemoglobin rate. So, the intraoperative transfusion rate amounted to 6% for the MECC group and 12.8% for the CPB group (P < .001). Patients in the CPB group had significantly higher levels of postoperative blood creatinine and urea. CONCLUSION: The MECC system is a new concept of CPB that seems to be reliable and safe. To perform CABG, the MECC provides an excellent surgical exposure like a standard CPB and a better biologic profile like CABG without CPB. PMID- 16368319 TI - Changing trends in the long-term prognosis of patients with acute myocardial infarction: a population-based perspective. AB - BACKGROUND: Little contemporary data exist describing changes over time in the postdischarge prognosis of hospital survivors of acute myocardial infarction (AMI). The purpose of our study was to examine recent and multidecade-long (1975 2001) trends in the long-term prognosis of greater Worcester (MA) residents discharged from all metropolitan Worcester hospitals after confirmed AMI. METHODS: A total of 9827 greater Worcester residents with independently validated AMI discharged from all metropolitan Worcester hospitals during 13 annual periods between 1975 and 2001 comprised the study population. A variety of follow-up approaches were used to ascertain the survival status of discharged patients through the end of 2003. RESULTS: Crude unadjusted 1-year postdischarge survival rates declined over time. Suggestions of modest improvements in the multivariable adjusted 1-year postdischarge survival rates were noted, however, in patients discharged from greater Worcester hospitals in the more recent (2001) (adjusted odds of surviving 1.23, 95% CI 0.97-1.55) as compared with earliest study periods (1975/1978). Advancing age, female sex, presence of prior diabetes, stroke, heart failure, or myocardial infarction and occurrence of several clinical complications during hospitalization were significantly related to an adverse postdischarge prognosis. CONCLUSIONS: The results of this investigation provide contemporary insights into the long-term survival of patients with AMI from a more generalizable population-based perspective. Multivariable adjusted analyses revealed slight improvements in postdischarge survival over time. Our data identify several high-risk groups that should be targeted for more aggressive surveillance and increased use of effective cardiac therapies and interventions. PMID- 16368320 TI - The cost of acute myocardial infarction in the new millennium: evidence from a multinational registry. AB - BACKGROUND: Although geographic variation in the treatment of acute myocardial infarction (AMI) has been recognized, the impact of evidence-based international treatment guidelines on such variation is unclear. We sought to characterize resource use and cost of initial hospitalization for AMI in 9 countries and compare the contribution of length of stay (LOS) and procedure use to cost. METHODS: We applied country-specific cost estimates to data from the international AMI registry associated with the VALIANT trial. The registry includes demographic, medical history, treatment, and discharge information for 5573 patients with AMI admitted to 84 sites in 9 countries from November 1999 to June 2001. Hospitalization cost was estimated by imputed discharge diagnosis related group code and adjusted for the LOS and procedures. Generalized linear regression was used to adjust cost by country; the contribution of LOS and procedures to cost was assessed by ordinary least squares regression. RESULTS: Unadjusted mean cost for initial AMI hospitalization was 9993 dollars (95% CI 9702 dollars-10,228 dollars). After adjustment for baseline patient-level variation, the lowest average cost was 1605 dollars (Argentina) and the highest was 9196 dollars (United States). Length of stay explained more of the variation in cost (50.7%) than did procedure intensity (31.9%). CONCLUSIONS: International differences in the cost of AMI persist, reflecting variations in procedure use and especially LOS. Relative differences in resource costs and incentives inherent in the provision and financing of health care likely contribute to treatment and cost variation and limit the widespread adoption of international practice guidelines. PMID- 16368321 TI - Quality of life in advanced heart failure: role of mitral regurgitation. AB - BACKGROUND: Quality of life is increasingly used as an important end point in clinical trials of treatments for heart failure; thus, relationships between traditional clinical variables and quality of life need to be understood. Baseline data from an ongoing multi-institutional study of a surgically implanted cardiac support device (CorCap, Acorn Cardiovascular, Inc, St Paul, MN) positioned around the heart to halt progression of remodeling in patients with cardiomyopathy provide an opportunity to study the relationship between mitral regurgitation (MR) and quality of life in a group of relatively young patients. OBJECTIVE: The objective of this study was to determine whether the degree of MR correlates with quality of life in patients presenting with significant symptoms of heart failure. METHODS: Baseline MR was assessed by echocardiography and patients were stratified according to whether there was a clinical indication for mitral valve surgery. The effect of heart failure on quality of life was measured by the Minnesota Living with Heart Failure questionnaire (MLHF). The New York Heart Association class, exercise performance measured by peak oxygen consumption and the 6-minute walk test, and the SF-36 physical function measure were analyzed as potential mediating variables. RESULTS: Mean MR grade was 2 +/- 1.5 on a 0-to 4 (worst) scale (n = 260) and ejection fraction averaged 27% +/- 9%. Most patients (82%) had New York Heart Association class III symptoms. Peak oxygen consumption averaged 14.7 +/- 4.3 mL/kg per minute and average walking distance was 348 +/- 83 m. Median (quartile range) SF-36 physical function was 35 (20-50) on a 0-to-100 (best) scale. Median MLHF score was 61 (47.5-77) on a 0-to-105 (worst) scale. The degree of MR and having an indication for mitral valve repair were not associated with the patients' quality of life. Controlling for symptoms and functional measures, older age was independently associated with better quality of life. CONCLUSIONS: The degree of MR was not related to MLHF scores, suggesting that surgery to reduce MR might not have predictable effects on quality of life. Further studies are needed to understand why younger patients reported worse quality of life and how this observation could impact therapy. PMID- 16368322 TI - Long-term cost-effectiveness of early and sustained clopidogrel therapy for up to 1 year in patients undergoing percutaneous coronary intervention after presenting with acute coronary syndromes without ST-segment elevation. AB - BACKGROUND: The superiority of clopidogrel and aspirin versus aspirin alone for up to 1 year in patients who undergo percutaneous coronary intervention (PCI) after presenting with acute coronary syndromes without ST-segment elevation was demonstrated in the PCI-CURE study. We evaluated the long-term cost-effectiveness of clopidogrel use for up to 1 year using patient-level outcomes and resource use from PCI-CURE, and estimates of life expectancy gains based on external sources. METHODS: PCI-CURE involved 2658 patients who underwent PCI between 1998 and 2000 after being randomized in the CURE trial to clopidogrel (n = 1313) or placebo (n = 1345). Roughly two thirds (clopidogrel n = 821, placebo n = 909) underwent PCI during the initial hospitalization (early PCI). Costs were applied to hospitalizations according to diagnosis-related group. Clopidogrel was assigned the average wholesale price of 3.22 dollars per day. Life expectancy gains resulting from the prevention of major clinical events were estimated using external sources. RESULTS: Average total costs were higher with clopidogrel (difference [based on costing method] 253 dollars-423 dollars). For patients who underwent PCI during the initial hospitalization, the difference ranged from 155 dollars lower to 90 dollars higher with clopidogrel. The estimated life expectancy gain with clopidogrel was 0.0885 years, whereas it was 0.0962 years for the early PCI subgroup. Incremental cost per year of life gained with clopidogrel ranges from 2856 dollars to 4775 dollars overall and from dominant (life expectancy benefit with cost savings) to 935 dollars for the early PCI subgroup. CONCLUSIONS: Clopidogrel given for up to 1 year in patients undergoing PCI after presentation with acute coronary syndromes is a highly cost-effective treatment strategy. PMID- 16368323 TI - Percutaneous versus surgical closure of secundum atrial septal defect: comparison of early results and complications. AB - BACKGROUND: Surgical closure of atrial septal defect (ASD) provides excellent results. Given the increasing popularity of percutaneous techniques, a comparison between the 2 methods is needed. METHODS: Between December 1988 and June 2003, we performed 1284 procedures in 1268 consecutive patients with isolated secundum ASD. Five hundred and thirty-three patients underwent surgical repair of ostium secundum ASD (group A). Seven hundred and fifty-one consecutive patients underwent percutaneous ASD closure (group B). The following outcomes were studied: mortality, morbidity, hospital stay, and efficacy. RESULTS: There were no postoperative deaths. The overall rate of complications was higher in group A than in group B: 44% (95% CI 39.8%-48.2%) versus 6.9 % (95% CI 5%-8.7%) (P < .0001). Major complications were also more frequent in group A: 16% (95% CI 13% 19%) versus 3.6% (95% CI 2.2%-5.0%) (P = .002). Multiple logistic regression analysis showed that surgery was independently strongly related to the occurrence of total complication (odds ratio [OR] 8.13, 95% CI 5.75-12.20) and of major complications (OR 4.03, 95% CI 2.38-7.35). The occurrence of minor complications was independently related to surgery (OR 7.33, 95% CI 4.75-11.02), childhood (OR 1.52, 95% CI 1.01-2.34), and presence of systemic hypertension (OR 1.35, 95% CI 1.01-4.41). Hospital stay was shorter in group B (3.2 +/- 0.9 vs 8.0 +/- 2.8 days, P < .0001). CONCLUSIONS: Percutaneous ASD closure provides, in experienced hands and in highly specialized centers, excellent results with a lower complication rate and requires a shorter stay in hospital. PMID- 16368324 TI - Immediate and late outcomes of patients undergoing transseptal left-sided heart catheterization for symptomatic valvular and arrhythmic diseases. AB - BACKGROUND: The transseptal technique has been widely used for diagnostic and therapeutic left-sided heart catheterization. However, its differential immediate and late outcomes among patients with various valvular and arrhythmic diseases are not yet determined. METHODS: Beginning from 1993, all patients undergoing transseptal procedures were screened and categorized into diagnosis, arrhythmia, and valvuloplasty groups according to the purposes of the catheterization. Incidences of transseptum-related acute major events (cardiac perforation, embolic stroke, and bradyarrhythmia during the procedure) and late complications (residual atrial septal defect [ASD], embolic stroke, bradyarrhythmia, and death up to 18 months) were analyzed and compared between groups. RESULTS: From January 1993 to May 2003, a total of 176 patients underwent 184 transseptal procedures for diagnosis of valvular heart diseases (n = 8), catheter ablation of arrhythmogenic foci (n = 29), and mitral valvuloplasty (n = 147). The immediate outcome was similar among the 3 groups, with an overall acute complication incidence of 3.8%. At follow-up, the incidences of bradyarrhythmia, embolic stroke, and death were not different among the 3 groups. Patients undergoing valvuloplasty had a significantly higher prevalence of residual ASD, especially for those with more severe mitral stenosis and less valvuloplasty success. However, presence of ASD did not impose disadvantage over the 1.5-year prognosis. CONCLUSION: Transseptal left-sided heart catheterization can be safely applied to patients with different categories of cardiac diseases with comparably good immediate and late outcomes. Although patients undergoing percutaneous valvuloplasty have a higher chance of permanent ASD creation, their prognosis is not influenced. PMID- 16368325 TI - Increased carotid and femoral intima-media thickness in patients after repair of aortic coarctation: influence of early repair. AB - BACKGROUND: In patients, after repair of aortic coarctation, abnormal function of the proximal precoarctation arterial conduits has been demonstrated, but data on arterial wall structure of proximal and distal arteries in patients are scarce. The aim of our study is to compare intima-media thickness (IMT) of the carotid and femoral arteries in controls and patients after coarctation repair and to identify independent predictors of carotid and femoral IMT. METHODS: A total of 137 patients after repair of aortic coarctation (89 male, median age 29.7 years, range 17-74 years, and median age at repair 6.3 years, range 0.01-46 years) and 46 age-matched controls underwent ambulatory blood pressure monitoring and B-mode ultrasound of the carotid and femoral arteries. RESULTS: Twenty (15%) of the 137 patients were operated on before the age of 1 year ("early repair"). The patients with an early repair were younger compared to the controls and compared with the patients with a late repair (24.5 [10.5] vs 31.6 [8.4] years, P = .005, and vs 32.1 [10.5] years, P < .001, respectively). Eighty patients (59%) had hypertension on the basis of the results of the ambulatory blood pressure monitoring (mean daytime blood pressure > or = 135/85 mm Hg). The prevalence of hypertension was not significantly different between patients with an early or late repair. Although patients with an early repair were younger, both carotid and femoral IMT were increased in these patients compared to controls (common carotid artery 0.57 [0.11] vs 0.49 [0.07] mm, P < .001; common femoral artery 0.48 [0.10] vs 0.44 [0.05] mm, P = .033). In multivariable regression analyses, age at repair was only an independent predictor of femoral and not of carotid IMT. CONCLUSIONS: Patients, after repair of aortic coarctation, have increased carotid and femoral IMT despite normal ambulatory blood pressures and successful previous repair. Early repair seems to preserve postcoarctation arterial wall structure, but seems to have only limited effect on precoarctation vessel walls, which supports the theory of a developmental defect of the proximal aorta and its branches. PMID- 16368326 TI - Implantation of a prosthesis mounted inside a self-expandable stent in the pulmonary valvar area without use of cardiopulmonary bypass. AB - Patients with right ventricular outflow tract reconstruction often require redo operations with time. Unique surgical problems exist in this group of patients. We report an innovative method of implantation of a pulmonary valve without the use of cardiopulmonary bypass. PMID- 16368327 TI - Homograft replacement of the calcified aortic root in familial hypercholesterolemia. AB - A 39-year-old woman with a known history of homozygous familial hypercholesterolemia was admitted with chest discomfort. Preoperative echocardiography and coronary angiography showed a heavily stenotic aortic valve as well as a calcific hypoplastic aortic root. Aortic root replacement using an aortic homograft was done. PMID- 16368328 TI - The impact of new technology on cardiothoracic surgical practice. PMID- 16368329 TI - Surgery for paroxysmal atrial fibrillation in the setting of mitral valve disease: a role for pulmonary vein isolation? AB - BACKGROUND: It is unknown whether pulmonary vein isolation or a complete Cox-Maze procedure is needed to ablate paroxysmal atrial fibrillation in patients with mitral valve disease. Our objective was to assess the impact of different surgical treatments for this arrhythmia in patients undergoing mitral valve surgery. METHODS: From July 1993 to January 2004, 152 patients underwent combined surgical treatment of paroxysmal atrial fibrillation and mitral valve disease. Ablation procedures included pulmonary vein isolation alone (n = 31, 20%), pulmonary vein isolation with left atrial connecting lesions (n = 80, 53%), and Cox-Maze (n = 41, 27%). The latter had longer durations of atrial fibrillation than the former (p < 0.0001). Rhythm documented on 1,225 postoperative electrocardiograms was used to estimate prevalence of, and risk factors for, atrial fibrillation across time. Ablation failure was defined as occurrence of atrial fibrillation any time beyond 6 months after operation. RESULTS: Prevalence of postoperative atrial fibrillation peaked at 22% at 2 weeks and declined to 9% at 1 year. Risk factors included older age (p = 0.09), larger left atrium (p = 0.05), and rheumatic (p = 0.003) and degenerative etiologies (p = 0.03). Freedom from ablation failure was 84% at one year. Ablation procedure did not affect prevalence of atrial fibrillation or incidence of ablation failure. CONCLUSIONS: Pulmonary vein isolation alone may be adequate treatment for patients with paroxysmal atrial fibrillation undergoing mitral valve surgery, particularly when it is of short duration. A randomized trial is necessary to examine this strategy, especially in patients with longer duration of paroxysmal atrial fibrillation. PMID- 16368330 TI - Long-term neurocognitive function after mechanical aortic valve replacement. AB - BACKGROUND: Mechanical aortic valves are a possible source of microemboli potentially causing cerebral injury. Therefore, the long-term impact of mechanical aortic valve replacement on neurocognitive function is uncertain. METHODS: In this prospective, contemporary study, we followed 32 consecutive patients (aged 51 +/- 8 years; range, 38 to 70; EuroSCORE [European System for Cardiac Operative Risk Evaluation] 4.4 +/- 1.7) undergoing isolated aortic valve replacement with a mechanical prosthesis. A cohort of age- and sex-matched patients (n = 28, aged 50 +/- 7 years) served as nonsurgical controls. After aortic valve replacement, neurocognitive function was serially reevaluated at 7 day (n = 32), 4-month (n = 31), and 3-year (n = 29) follow-up. Neurocognitive function was measured by means of P300 auditory evoked potentials. RESULTS: Before the operation, P300 peak latencies were comparable between surgical patients (361 +/- 32 ms) and nonsurgical controls (365 +/- 33 ms, p = 0.783). In patients undergoing aortic valve replacement, P300 peak latencies were prolonged 7 days after surgery (380 +/- 32 ms) as compared with before the operation (361 +/- 32 ms, p < 0.0001) and as compared with nonsurgical controls (364 +/- 34 ms, p = 0.002). At 4-month (369 +/- 30 ms, p = 0.752) and 3-year (370 +/- 31 ms, p = 0.825) follow-up, P300 peak latencies normalized as compared with before operation and as compared with nonsurgical controls (4-month follow-up 363 +/- 31 ms, p = 0.832; 3-year follow-up 366 +/- 32 ms, p = 0.432). We found no difference in patients with different valve types. CONCLUSIONS: Despite previous assumptions based on the potential occurrence of microemboli in patients with mechanical valves, mechanical aortic valve replacement has no adverse long-term impact on neurocognitive function. This finding is only valid for patients with a comparable age range undergoing isolated aortic valve replacement. PMID- 16368331 TI - Intraoperative and postoperative evaluation of cavitation in mechanical heart valve patients. AB - BACKGROUND: Cavitation has been claimed partly responsible for the increased risk of thromboembolic complications, hemolysis, and fatal valve failure seen in mechanical heart valve patients. In vivo studies have investigated cavitation using high-pass filtering of the high-frequency pressure fluctuations with the root mean square values as an assessment of intensities. In vitro studies have shown that this well-known method may not be ideal owing to loss of data as a consequence of filtering, and because it requires a priori knowledge of the valve resonance pattern. Therefore, a new method has been developed, which decomposes the signal into nondeterministic (cavitation) and deterministic (valve resonance) signal components, and hence decreases data loss. This study aimed to evaluate cavitation in patients with mechanical, biological, and native heart valves both intraoperatively and postoperatively using the new method. METHODS: High frequency pressure fluctuations were measured by a hydrophone intraoperatively and postoperatively in 14 patients with mechanical valves, 10 patients with normal aortic valves, and 5 patients with bioprosthesis. The total signal energy was evaluated as nondeterministic and deterministic energies. RESULTS: Nondeterministic energies were verified both intraoperatively and postoperatively in all patients who had a mechanical valve; this finding confirms the cavitation potential of mechanical valves. None of the data recorded in patients with bioprosthetic or native valves contained nondeterministic energy. CONCLUSIONS: The study confirms the presence of cavitation in mechanical heart valve patients using the nondeterministic energy of high-frequency pressure fluctuations as a quantitative measure of cavitation both intraoperatively and postoperatively. PMID- 16368332 TI - Coapsys mitral annuloplasty for chronic functional ischemic mitral regurgitation: 1-year results. AB - BACKGROUND: Untreated functional ischemic mitral regurgitation (MR) leads to reduced survival in patients undergoing coronary artery bypass grafting (CABG). However, mitral repair or replacement increases mortality and morbidity over CABG alone. The Myocor Coapsys annuloplasty system potentially reduces these risks by facilitating MR reduction on a beating heart without atriotomy. We present data from the first 11 patients completing 1-year follow-up of a total of 34 implanted patients. METHODS: Patients referred for CABG with preoperative grade 2 or greater ischemic functional MR were included in this study. Patients with structural valve defects or who demonstrated MR less than grade 2 after CABG, despite hemodynamic challenge, were intraoperatively excluded. Coapsys consists of two epicardial pads connected by a flexible chord implanted by passing the chord across the left ventricle with special instruments without cardiopulmonary bypass. The system was sized to reduce critical valve dimensions and MR. Serial clinical and echocardiographic data were collected out to 1 year. RESULTS: Mean age was 58.1 +/- 6.6 years and mean ejection fraction, 38.5% +/- 7.1%. From baseline to 1-year follow-up, effect on MR grade, MR jet area (cm2), and New York Heart Association class were, respectively, 2.9 +/- 0.5 to 1.1 +/- 0.8, 7.4 +/- 2.9 to 3.0 +/- 1.6, and 2.5 +/- 0.5 to 1.2 +/- 0.4 (all p < 0.05 versus baseline). During follow-up, there were no deaths, device failures, reemergence of grade 3 or 4 MR, heart failure readmission, or valve reoperations. CONCLUSIONS: The Coapsys annuloplasty system is effective in reducing functional ischemic MR and improving NYHA class. The initial data are encouraging and suggest that the device is safe and benefits are sustained at 1 year. PMID- 16368333 TI - Cholesterol-modified polyurethane valve cusps demonstrate blood outgrowth endothelial cell adhesion post-seeding in vitro and in vivo. AB - BACKGROUND: The clinical and experimental use of polyurethane heart valve prostheses has been compromised by thrombosis and calcified thrombus. This is caused in part by the lack of an intact endothelium on these implant surfaces. We hypothesize that endothelial seeding of a polyurethane heart valve leaflet with autologous sheep blood outgrowth endothelial cells (BOECs) could be achieved with cholesterol-modified polyurethane (PU-Chol) to promote BOEC adhesion, thereby resulting in an intact, shear-resistant endothelium that would promote resistance to thrombosis. METHODS: Cholesterol-derivatized polyurethane was formulated by bromoalkylation of the urethane nitrogens followed by reactive attachment of mercaptocholesterol. In vitro shear flow studies were carried out comparing BOEC retention on control surfaces versus PU-Chol using forces comparable to those observed in vivo with cardiac valves (75 dyne/cm2). Autologous sheep BOECs were seeded onto PU-Chol before pulmonary leaflet replacement surgery under cardiopulmonary bypass. Studies were terminated at 30 and 90 days followed by retrieval analyses. RESULTS: Blood outgrowth endothelial cell seeding of PU-Chol surfaces resulted in an endothelial monolayer that was positive for von Willebrand factor. Polyurethane-cholesterol demonstrated significantly greater BOEC adhesion under 75 dyne/cm2 shear force in vitro than control polyurethane (75.3% +/- 12.3% versus 5.8% +/- 3.9%, respectively; p < 0.001). Sheep pulmonary cusp replacements demonstrated retention of seeded BOECs on PU-Chol leaflets with no significant differences in the extent of cellular density comparing unimplanted specimens with explants. Control explants (nonseeded PU-Chol and nonseeded polyurethane) demonstrated no evidence of endothelial recruitment. CONCLUSIONS: Polyurethane-cholesterol represents a polyurethane formulation with very high adhesive properties for BOECs under heart valve level shear forces both in vitro and in vivo. PMID- 16368334 TI - Invited commentary. PMID- 16368336 TI - Invited commentary. PMID- 16368335 TI - In vitro fabrication of a tissue engineered human cardiovascular patch for future use in cardiovascular surgery. AB - BACKGROUND: One approach to tissue engineering has been the development of in vitro conditions for the fabrication of functional cardiovascular structures intended for implantation. In this experiment, we developed a pulsatile flow system that provides biochemical and biomechanical signals in order to regulate autologous, human patch-tissue development in vitro. METHODS: We constructed a biodegradable patch scaffold from porous poly-4-hydroxy-butyrate (P4HB; pore size 80 to 150 microm). The scaffold was seeded with pediatric aortic cells. The cell seeded patch constructs were placed in a self-developed bioreactor for 7 days to observe potential tissue formation under dynamic cell culture conditions. As a control, cell-seeded scaffolds were not conditioned in the bioreactor system. After maturation in vitro, the analysis of the tissue engineered constructs included biochemical, biomechanical, morphologic, and immunohistochemical examination. RESULTS: Macroscopically, all tissue engineered constructs were covered by cells. After conditioning in the bioreactor, the cells were mostly viable, had grown into the pores, and had formed tissue on the patch construct. Electron microscopy showed confluent smooth surfaces. Additionally, we demonstrated the capacity to generate collagen and elastin under in vitro pulsatile flow conditions in biochemical examination. Biomechanical testing showed mechanical properties of the tissue engineered human patch tissue without any statistical differences in strength or resistance to stretch between the static controls and the conditioned patches. Immunohistochemical examination stained positive for alpha smooth muscle actin, collagen type I, and fibronectin. There was minor tissue formation in the nonconditioned control samples. CONCLUSIONS: Porous P4HB may be used to fabricate a biodegradable patch scaffold. Human vascular cells attached themselves to the polymeric scaffold, and extracellular matrix formation was induced under controlled biomechanical and biodynamic stimuli in a self-developed pulsatile bioreactor system. PMID- 16368337 TI - Surgery for ventricular tachycardia in patients undergoing left ventricular reconstruction by the Dor procedure. AB - BACKGROUND: Surgical ventricular restoration (the Dor procedure) is an option in patients with coronary artery disease and postinfarction left ventricular aneurysm. The procedure can be extended to treat ventricular tachycardia (VT). The aim of this study was to evaluate the Dor procedure including VT surgery in our institution. METHODS: From July 1997 to December 2003, 53 consecutive patients with left ventricular aneurysm and VT underwent surgical ventricular restoration including nonguided endocardiectomy and cryoablation. Twenty-four patients had at least one preoperative episode of spontaneous VT, of which 8 were survivors of sudden cardiac death. Twenty-nine patients had inducible-only VT. In 45 patients, who underwent preoperative programmed stimulation, sustained uniform VT could be initiated. Arrhythmia control was evaluated by programmed stimulation or analysis of events registered by implanted defibrillators and by review of patient's records. RESULTS: Early mortality was 2 of 53 (3.8%). Mean follow-up was 3.7 years. At 1, 3, and 5 years overall actuarial survival was 94%, 80%, and 59%, respectively. Surgical success rate in patients with preoperative spontaneous VT was 91%. Inducible VT was found in 5 of 35 patients who underwent postoperative programmed stimulation. There was no arrhythmia-related late death and there was no loss to follow-up. CONCLUSIONS: The Dor procedure including VT surgery is an effective treatment for postinfarction left ventricular aneurysm and VT and eliminates the need for an implantable defibrillator in most patients. Early and long-term results are good in terms of survival and arrhythmia control. PMID- 16368338 TI - Microwave ablation for atrial fibrillation: dose-response curves in the cardioplegia-arrested and beating heart. AB - BACKGROUND: Microwave ablation has been used to replace the traditional incisions used in the surgical treatment of atrial fibrillation. However, dose-response curves have not been established in surgically relevant models. The purpose of this study was to develop dose-response curves for the Flex 10 (Guidant, Inc) microwave device in both the acute cardioplegia-arrested heart and on the beating heart. METHODS: Twelve domestic pigs (40 to 45 kg) were subjected to microwave ablation in either the arrested (n = 6) or beating heart (n = 6). The cardioplegia-arrested heart was maintained at 10 degrees to 15 degrees C while six atrial endocardial and seven right ventricular epicardial lesions were created in each animal. On the beating heart, six right atrial and seven ventricular epicardial lesions were created. Ablations were performed for 15, 30, 45, 60, 90, 120, and 150 seconds (65 W, 2.45 GHz). The tissue was stained with 2,3,5-triphenyl-tetrazolium chloride, and sectioned at 5-mm intervals. Lesion depth and width were determined from digital micrographs. RESULTS: Mean atrial wall thickness was 2.8 mm (range, 1 to 8 mm). In the arrested heart, 94% of atrial lesions were transmural at 45 seconds and 100% were transmural at 90 seconds. In the beating heart, only 20% of atrial lesions were transmural despite prolonged ablation times (90 seconds). Ventricular lesion width and depth increased with duration of application, and were similar on the arrested and beating hearts. CONCLUSIONS: Microwave ablation produces linear dose-response curves. Transmural lesions can be reliably produced on the arrested heart, but not consistently on the beating heart. PMID- 16368339 TI - Invited commentary. PMID- 16368340 TI - Preoperative statin therapy and troponin T predict early complications of coronary artery surgery. AB - BACKGROUND: Pretreatment with statins reduces early ischemic events after percutaneous coronary interventions, primarily in patients with a high level of inflammation markers. We sought to examine the association between preoperative statin therapy, systemic inflammation, and myocardial ischemia with the occurrence of early cardiac complications after coronary artery bypass grafting surgery. METHODS: One hundred forty-one consecutive patients who underwent coronary artery bypass grafting surgery from two university tertiary hospitals were stratified according to their preoperative status of statin therapy (87 treated and 54 nontreated). Preoperative blood samples were collected for measurement of lipid parameters, C-reactive protein, interleukin-6, and troponin T. The evaluated primary endpoint was a composite of death and myocardial infarction at 30 days. RESULTS: Patients undergoing preoperative statin therapy showed a reduced incidence of death (2.3% versus 13.0%, p = 0.012), myocardial infarction (5.7% versus 18.5%, p = 0.017), and primary combined endpoint (8.0% versus 22.2%, p = 0.017). In the multivariate model, preoperative troponin T greater than 0.01 ng/mL (odds ratio 6.85, p = 0.001) and nonstatin therapy (odds ratio 4.2, p = 0.01) predicted a higher risk of primary endpoint. Statins showed a significant interaction with troponin T status and benefited primarily those patients with positive troponin T. Among 19 patients with troponin T greater than 0.01 ng/mL, the primary endpoint occurred in all 6 nonstatin-treated patients, but it occurred in only 1 of 13 statin-treated patients (p < 0.001). Neither C reactive protein nor interleukin-6 predicted early complications, nor did they interact with statin therapy (p = not significant). CONCLUSIONS: Preoperative statin therapy reduces early complications and offers additional protection in patients with positive troponin T status, regardless of inflammatory markers. PMID- 16368341 TI - Invited commentary. PMID- 16368342 TI - Off-pump myocardial revascularization: critical analysis of 23 years' experience in 3,866 patients. AB - BACKGROUND: Surgical myocardial revascularization without cardiopulmonary bypass (CPB) is not new, with the first consecutive series of patients appearing in the early eighties. There has been increased interest in this alternative approach, especially in patients with comorbidities. There is controversy regarding advantages, risks, and usefulness of this method of myocardial revascularization. We herein report a consecutive series of 3,866 patients, from the first case in September 1981 to the last in November 2004, analyzing applicability, mortality, morbidity, and surgical technique. METHODS: From September 1981 to November 2004, 3,866 consecutive patients were revascularized without CPB. This figure represents an overall applicability of 30.8% considering a total of 12,553 revascularization procedures performed during this time. There were 2,822 males (73%) with ages from 12 to 93 years (median, 62 +/- 14). Mean grafts per patient was 1.9, and the internal mammary artery was used in 87.3% of cases. The main indications for surgery were chronic coronary insufficiency (89% of cases) and failure of angioplasty or stenting. RESULTS: Hospital mortality was 1.9%, with low incidence of cerebrovascular accident (5 cases in the entire series). Morbidity, considering major postoperative complications, occurred in 12.5% of the patients. The applicability of the off-pump technique was 18% of cases in the beginning of our experience, increasing to 49% in the last 5 years with the use of stabilizers and maneuvers to expose posterior coronary branches. CONCLUSIONS: Off-pump coronary surgery is an alternative method of myocardial revascularization that should be considered for every patient. The preference of this technique over conventional revascularization should be based on the surgeon's own experience, on the patient's preoperative condition and on the coronary anatomy. Off-pump myocardial revascularization represents an important development in coronary artery surgery. Over the years it has evolved into a valid form of surgery with the same safety as the conventional operation and with more advantages in high risk patients. PMID- 16368343 TI - Six-month angiographic follow-up of the PAS-Port II clinical trial. AB - BACKGROUND: The PAS-Port device (Cardica, Redwood City, CA) allows the rapid deployment of a clampless proximal anastomosis between a vein graft and the aorta. METHODS: Fifty-four patients awaiting elective coronary artery bypass graft surgery were enrolled. Outcome variables were intraoperative device performance, early and 6- month angiographic graft patency, and 12-month clinical follow-up. RESULTS: Sixty-three PAS-Port devices were deployed in 54 patients. Two deployments were unsuccessful. There were no reoperations for bleeding. Two patients died of causes unrelated to the device. Patency evaluation at discharge was performed by angiogram on 49 implants and computed tomography in 2 implants (86% follow-up). At discharge, all evaluated grafts were patent (100%) and rated Fitzgibbon A. At 6-month follow-up, there was no additional mortality; 47 implants (88% follow-up) were evaluated by angiography (Fitzgibbon O [n = 1], Fitzgibbon B [n = 1], and Fitzgibbon A [n = 45]) and 5 by computed tomography. All grafts but 1 were patent (98.1%). At 12 months, 2 additional patients died of causes unrelated to the PAS-Port implant. Forty-six of 50 alive patients (95.8%) were followed up without any reports of device-related major adverse cardiac events. CONCLUSIONS: Discharge (100%) and 6-month patency (98%) are excellent; patency and 12 months' clinical follow-up compares favorably with data from historical hand-sewn controls. The PAS-Port system safely allows the clampless creation of a proximal anastomosis. PMID- 16368344 TI - Splanchnic organ injury during coronary surgery with or without cardiopulmonary bypass: a randomized, controlled trial. AB - BACKGROUND: We investigated the efficacy of coronary surgery with or without cardiopulmonary bypass in protecting the function of the small intestine, liver, and pancreas. METHODS: Patients were randomized to off-pump coronary artery bypass grafting (OPCAB) or coronary artery bypass grafting with cardiopulmonary bypass (CABG-CPB). Small intestine function was assessed by differential four sugars (O = methyl-D-glucose, D-xylose, L-rhamnose, and lactulose) permeability and absorption tests. Liver function was assessed by monoethylglycinexylidide/lidocaine ratios and by serial measurements of transaminases (aspartate transaminase and alanine-amino transferase), bilirubin, and alkaline phosphatase. Pancreatic function was assessed by serial measurements of insulin/glucagon ratio, amylase, and glucose. Forty patients were recruited (20 per group). RESULTS: Permeability and absorption were more impaired in the OPCAB group immediately after surgery, but returned to baseline levels in both groups by postoperative day 5 (interaction of surgery type and time; p = 0.05 and p = 0.02, respectively). Monoethylglycinexylidide/lidocaine ratios were not different in the two groups. Aspartate transaminase and alanine-amino transferase levels were higher in the CABG-CPB group for the first postoperative day, but levels converged by day 3 (interaction of surgery type and time; p < 0.0001 and p = 0.04, respectively). The bilirubin level for the OPCAB group overshot the CABG CPB group at 36 hours before returning to a similar level 60 hours postoperatively. Amylase levels were higher in the CABG-CPB group than in the OPCAB group (1.17 times; p = 0.03); other markers of pancreatic function showed no differences between the groups. CONCLUSIONS: Early small intestine function is worse with OPCAB; all functions recover to similar levels in both groups by day 5. Conversely, pancreatic function is worse with the CABG-CPB group than with the OPCAB group. Hepatic metabolic function does not differ by type of surgery to the end of the operation. Postoperative hepatocellular injury was worse with the CABG CPB group. PMID- 16368345 TI - Aprotinin shows both hemostatic and antithrombotic effects during off-pump coronary artery bypass grafting. AB - BACKGROUND: Hemostatic drugs are widely thought to be unnecessary and potentially detrimental in off-pump coronary artery bypass graft surgery (OPCABG), despite well-established use in on-pump surgery. In a randomized, prospective OPCABG trial, we assessed efficacy and safety of aprotinin through a comprehensive assessment of graft patency and hematologic function. METHODS: Sixty patients were randomly assigned to full-dose aprotinin or placebo. Heparin was titrated to a kaolin-based activated clotting time of greater than 300 seconds. Exclusionary criteria included creatinine greater than 2 mg/dL, conversion to on-pump CABG, and preoperative GPIIb/IIIa inhibition. Hematologic assessments were obtained preoperatively, at the end of surgery, and on days 1 and 3: mean platelet volume, thrombin generation (prothrombin fragment 1.2 assay), and aspirin resistance using a modified thrombelastography, whole blood aggregometry, 11-dehydro thromboxane B2 levels, and flow cytometry. Thrombotic events were defined as postoperative myocardial infarction by electrocardiography or elevated troponin I, clinical stroke by examination and head computed tomography, and bypass graft failure by multichannel computed tomography angiography on day 5. RESULTS: Aprotinin was associated with a significant reduction in intraoperative and postoperative blood loss compared with placebo but had no effect on transfusion rates. Patients treated with aprotinin had significantly fewer thrombotic events (3% versus 23%, p < 0.05, Fisher's exact test) and less postoperative aspirin resistance (20% versus 46%, respectively, p < 0.05, Fisher's exact test). Postoperative prothrombin fragment 1.2 level was reduced by aprotinin use. CONCLUSIONS: Aprotinin reduced perioperative bleeding after OPCABG. Preserved aspirin sensitivity in the aprotinin group may explain the observed reduction in thrombotic events and might be related to the suppression of perioperative and transmyocardial thrombin formation. PMID- 16368346 TI - Radial artery angiographic string sign: clinical consequences and the role of pharmacologic therapy. AB - BACKGROUND: The radial artery is an increasingly important graft for coronary artery bypass surgery. Postoperative angiographic studies have shown that a proportion of radial grafts become diffusely narrowed but not occluded, or string signs. METHODS: Four hundred forty patients receiving a radial artery graft enrolled in a large clinical trial underwent postoperative angiography at 1 year. Angiograms were analyzed visually and quantitatively. A complete string sign was defined as diffuse narrowing along the full length of the graft, while a partial string sign was defined as segmental narrowing. Angiographic findings were correlated with medication compliance and clinical sequelae. RESULTS: Thirty-one patients (7.0 %) had radial artery graft string signs versus 4 patients (0.9%) with a saphenous vein graft string sign (p = 0.001). Complete string signs were present in 28 cases, and the mean diameter was 0.76 +/- 0.14 mm (mean +/- SD), whereas 3 cases had a partial string sign with a diameter of 0.89 +/- 0.14 mm. Fifteen radial arteries showed Thrombolysis in Myocardial Infarction Study (TIMI) 1 flow, 3 cases showed TIMI 2 flow, and 13 cases showed TIMI 3 flow. There was no difference in incidence of radial string sign between patients taking nifedipine versus diltiazem postoperatively. Multivariate analysis revealed the presence of radial artery string sign was closely related to the perioperative use of alpha adrenergic agonists and target vessels stenosis less than 90%. Postoperative symptoms were associated with radial artery string signs with TIMI 1 flow (p = 0.0045). CONCLUSIONS: In the Radial Artery Patency Study, radial artery string sign was present in 7% of patients. Despite diffuse narrowing, 52% of grafts had TIMI 2 flow or better. PMID- 16368347 TI - Effects of papaverine and glycerylnitrate-verapamil solution as topical and intraluminal vasodilators for internal thoracic artery. AB - BACKGROUND: Arterial spasm is described as an event occurring after left internal thoracic artery (LITA) harvesting. Many vasodilators have been used to treat or prevent LITA spasm. The aim of this study is to compare the effects between glyceril-trinitrate/verapamil (GV) solution and papaverine to treat LITA spasm and to determine the best delivery method. METHODS: One hundred consecutive ischemic patients were randomly assigned to three groups: group GV (n = 34, GV solution), group P (n = 33, papaverine), or group C (n = 33, normal saline). In each patient, pedicled LITA was harvested, thereafter sprayed with the randomized solution, and covered with a sponge. Fifteen minutes after heparin administration, LITA was distally divided; flow per minute was calculated after measuring the free flow for over 15 seconds; this is named "topical free flow." Then, the vasodilator was injected intraluminally and free flow per minute was measured; this is called "intraluminal free flow." RESULTS: Analysis of variance was applied to detect differences among groups; paired-sample t test was used for LITA topical free flow versus intraluminal free flow within single groups. Mean LITA free flows were as follows: group GV, topical free flow = 38.6 +/- 25.2 mL/min versus intraluminal free flow = 58.8 +/- 29 mL/min (p < 0.0001); group P, topical free flow = 45.4 +/- 38.9 mL/min versus intraluminal free flow mL/min (p < 0.0001); group C, topical free flow = 31.6 +/- 19.9 mL/min versus intraluminal free flow = 34 +/- 19.8 mL/min (p = 0.14). Topical free flow difference among the three groups was not statistically significant (p = 0.1); intraluminal free flow difference was statistically significant (p = 0.001). Intraluminal free flow in group GV and in group P were higher than intraluminal free flow in group C (p = 0.004 and 0.001, respectively). Intraluminal free flow of group P was higher than that of group GV; this difference did not reach statistical significance (p = 1.00). CONCLUSIONS: Glyceril-trinitrate/verapamil solution and papaverine are able to treat the spasm and increase the flow of the LITA, when they are used intraluminally. When used topically, these vasodilator agents do not ensure an optimal free flow. PMID- 16368348 TI - Comparative effects of tolazoline and nitroprusside on human isolated radial artery. AB - BACKGROUND: The radial artery is increasingly being used in coronary revascularization as an alternative conduit to a saphenous vein graft. Its perfect endothelial capacity provides a high patency rate comparable with the internal mammary artery (IMA). However, its spastic characteristics cause difficulties during its intraoperative preparation and may lead to early postoperative graft failure. Thus, treatment and/or prevention of radial artery spasm with an effective vasodilator agent is essential for its longevity. Endogenous vasoconstrictors, including noradrenaline, endothelin-1, and thromboxane A2, are likely to play a role in the pathogenesis of graft spasm. In the present study, we evaluated the vasorelaxant effect of tolazoline, a nonselective alpha-adrenoceptor blocker, against the contractions induced by various spasmogenic agents in an isolated human radial artery. METHODS: Tolazoline (10(-9)-10(-4) M) or sodium nitroprusside (SNP, 10(-9)-10(-4) M) were cumulatively applied on radial artery rings precontracted submaximally with noradrenaline, endothelin-1, thromboxane analogue, U46619, or potassium chloride. In addition, some rings were pretreated with tolazoline (4 x 10(-6) M) for 30 minutes and the contractile response curve to noradrenaline was assessed in its presence. RESULTS: Tolazoline effectively reversed noradrenaline-induced contractions in the radial artery, whereas it failed to produce remarkable relaxations on rings contracted with other spasmogenic agents, while SNP overcame the contractions induced by all spasmogens to a similar extent. In addition, brief pretreatment of radial artery rings with tolazoline significantly inhibited the contractions to noradrenaline. CONCLUSIONS: Tolazoline is not as broadly effective as SNP against all spasmogens investigated; however, it may be effective in counteracting alpha-adrenoceptor-mediated vasospasm in human radial arteries. PMID- 16368349 TI - Tumor necrosis factor alpha influences the inflammatory response after coronary surgery. AB - BACKGROUND: A systemic inflammatory response is not uncommonly observed after coronary revascularization. Tumor necrosis factor alpha is one of a number of modulators of this response. A functional polymorphism within the TNFalpha gene at position G-308A has been associated with increased TNFalpha levels. The relationship between predicted TNFalpha genotype and circulating TNFalpha levels in patients undergoing coronary revascularization surgery has yet to be defined. We examined the relationship between TNFalpha G-308A polymorphism, TNFalpha postoperative levels, and clinical outcome after coronary revascularization surgery. METHODS: We obtained DNA from 96 consecutive patients who underwent elective coronary revascularization. Patients were genotyped for TNFalpha G-308A polymorphism using sequence specific primer-polymerase chain reaction (SSP-PCR). Tumor necrosis factor alpha levels were measured on serum samples taken 3 hours postoperatively using enzyme-linked immunosorbent assay (ELISA). RESULTS: The prevalence of AA, AG, and GG TNFalpha-308 genotype was 12%, 40%, and 48%, respectively. Patients homozygous for A had higher circulating levels of TNFalpha (p = 0.009). Higher levels of TNFalpha were significantly associated with prolonged intensive care unit stay (p = 0.008), increase usage of an inotropic agent (p = 0.024), increased mortality risk (p = 0.018), and diabetes (p = 0.019). These remained statistically significant after risk stratification. CONCLUSIONS: Patients of the AA-308 TNFalpha genotype showed significantly higher TNFalpha plasma levels. Higher plasma levels of TNFalpha were associated with less favorable outcome after coronary revascularization surgery. It may prove useful to utilize TNFalpha serum levels as a marker for identifying high-risk patients in the future. PMID- 16368350 TI - Invited commentary. PMID- 16368351 TI - Benefit of partial right-bilateral internal thoracic artery harvesting in patients at risk of sternal wound complications. AB - BACKGROUND: Excellent long-term patency of the internal thoracic artery (ITA) graft promotes use of bilateral ITA bypass grafting; sternal devascularization, however, increases the risk of wound complications. We hypothesized that restricting right ITA (RITA) harvesting to a short proximal skeletonized segment (3 to 5 cm) would result in adequate residual blood supply to reduce that risk. METHODS: Seventy-eight patients with numerous risk factors for wound complications underwent composite double ITA grafting, utilizing the RITA segment anastomosed to the left skeletonized ITA and to the obtuse marginal branch in Y fashion. Blood flow in the distal RITA was assessed by parasternal transthoracic Doppler ultrasonography. Comparisons were made with prospectively collected data of patients undergoing pedicled single (n = 160) or skeletonized bilateral ITA grafting (n = 143) during the same period. RESULTS: Incidence of obesity, chronic obstructive pulmonary disease, diabetes, and peripheral vascular disease was higher in study patients. Postoperative Doppler ultrasonography detected reversed systolic dominant flow pattern. Wound complications occurred in 2 of 78 (2.6%) patients, compared with 14 of 143 (9.8%) after bilateral ITA (p = 0.04) and 8 of 160 (5%) after single ITA grafting (p = ns). Technique of bilateral ITA harvesting (partial right versus full length; odds ratio, 0.2; confidence interval: 0.04 to 0.9) and diabetes mellitus (odds ratio, 2.7; 95% confidence interval: 1.1 to 6.3) were independent predictors of wound complications in the entire series. CONCLUSIONS: Substantial residual blood supply is detectable after partial RITA harvesting and may prevent wound complications in high-risk patients. PMID- 16368352 TI - Invited commentary. PMID- 16368353 TI - Upper extremity arteriovenous fistulas induce modest hemodynamic effect on the in situ internal thoracic artery. AB - BACKGROUND: The left internal thoracic artery is the ideal conduit for coronary artery revascularization due to superior patency compared with venous grafts. The hemodynamic effects of an arterio-venous fistula on the native in situ internal thoracic artery and the potential for coronary flow steal after revascularization with the internal thoracic artery, are not well-characterized. METHODS: Fifteen chronic hemodialysis patients with functioning left upper extremity arterio venous fistulas were evaluated with the use of transthoracic color Doppler analysis. Flow characteristics of the left and right internal thoracic arteries at baseline, with arterio-venous fistula occluded, and during hemodialysis were recorded. Peak systolic and diastolic velocities for the right and left internal thoracic arteries were calculated for each fistula state (occluded, open, and on dialysis). One-way analysis of variance was used to compare the different means. RESULTS: Mean flow velocity calculations failed to identify any statistically significant differences between the ipsilateral and contralateral internal thoracic artery in any fistula state. In addition, mean internal thoracic flow velocities were similar between the different fistula states. CONCLUSIONS: Changes in arteriovenous fistula flow state did not significantly alter Doppler flow hemodynamics of either the ipsilateral or contralateral in-situ internal thoracic artery. Further studies of the possible effects on internal thoracic arteries used as pedicled coronary grafts may be required. PMID- 16368354 TI - Role of the sarcolemmal adenosine triphosphate-sensitive potassium channel in hyperkalemic cardioplegia-induced myocyte swelling and reduced contractility. AB - BACKGROUND: Hyperkalemic cardioplegia (Plegisol) has been shown to result in myocyte swelling and reduced contractility. We have demonstrated the elimination of these detrimental effects by the addition of an adenosine triphosphate sensitive K+ (KATP) channel opener. To examine whether the mitochondrial or sarcolemmal KATP channel might be involved, volume and contractility in isolated myocytes from wild-type mice and mice lacking the sarcolemmal KATP channel (Kir6.2-/-) were evaluated. METHODS: Myocytes were perfused for 20 minutes each with control 37 degrees C Tyrode's solution, test solution, and then control solution. Test solutions were (n = 10 per group) either 9 degrees C Plegisol or 9 degrees C Plegisol with 100 micromol/L of diazoxide, a putative mitochondrial specific KATP channel opener. Cell volume and contractility were measured by digital video microscopy at baseline and during the test solution and reexposure periods. RESULTS: Myocytes from wild-type mice, perfused with 9 degrees C Plegisol, demonstrated significant cell swelling (11.2% +/- 0.4%; p < 0.01) and diminished contractility (32.5% +/- 9.6% reduction in percent shortening, 47.2% +/- 10.1% reduction in peak velocity of shortening, and 52.0% +/- 8.8% reduction in peak velocity of relengthening; p < 0.05) versus baseline. Cell swelling and diminished contractility were significantly reduced by the addition of diazoxide. In Kir6.2-/- myocytes, Plegisol caused a greatly reduced level of cell swelling (3.2% +/- 0.1%; p < 0.01), and this was unaffected by diazoxide. Contractility was unchanged in Kir6.2-/- myocytes after Plegisol. CONCLUSIONS: The sarcolemmal KATP channel appears necessary for exaggerated cell swelling and reduced contractility to occur after hyperkalemic cardioplegia in mouse myocytes. PMID- 16368355 TI - Hyperkalemic cardioplegia-induced myocyte swelling and contractile dysfunction: prevention by diazoxide. AB - BACKGROUND: Hyperkalemic cardioplegia (9 degrees C) results in significant myocyte swelling and reduced contractility, representing a possible mechanism of myocardial stunning. Adenosine triphosphate-sensitive potassium channel (KATP) openers have been shown to ameliorate stunning. This study evaluated the hypothesis that a KATP opener would prevent hyperkalemic cardioplegia-induced myocyte swelling and reduced contractility. METHODS: Isolated rabbit myocytes were perfused with 37 degrees C Tyrode's solution for 20 minutes, followed by test solution (9 degrees C or 37 degrees C) including control Tyrode's, Tyrode's + 100 micromol/L diazoxide (KATP opener), St. Thomas's solution; or 9 degrees C St. Thomas's + 100 micromol/L diazoxide or St. Thomas's + 100 micromol/L diazoxide + 20 micromol/L HMR1098 or 50 micromol/L 5-hydroxydeconoate (KATP blockers) for 20 minutes (n = 8 per group). Myocytes were then reexposed to 37 degrees C Tyrode's solution for 20 minutes. Volume and contractility were measured by videomicroscopy and video-based edge detection, respectively. RESULTS: St. Thomas's solution (9 degrees C) caused significant myocyte swelling and associated reduced contractility (p < 0.05). The addition of diazoxide abolished myocyte swelling (p < 0.0001), and eliminated the associated reduced contractility (p < 0.05). Findings were unchanged by the addition of HMR 1098 and 5-hydroxydeconoate. CONCLUSIONS: Diazoxide prevented myocyte swelling and reduced contractility secondary to hyperkalemic cardioplegia, and this was unchanged by the addition of either KATP channel blocker. Prevention of myocyte swelling was associated with improved contractility, consistent with the hypothesis that myocyte swelling may be a mechanism of myocardial stunning. Diazoxide may play a role in myocyte volume homeostasis by means of a mechanism separate from opening the KATP channel. PMID- 16368356 TI - Hybrid surgical angiogenesis: omentopexy can enhance myocardial angiogenesis induced by cell therapy. AB - BACKGROUND: The conditions at the injection site are important in cell transplantation for severe ischemic heart disease. The omentum is both a well vascularized tissue and a source of angiogenic factors. We examined the effectiveness of autologous bone marrow-derived mononuclear cells (BM-MNCs) with or without omentopexy in a large animal model. METHODS: Myocardial infarction was generated in the lateral wall by ligation of coronary artery branches in miniswine. Animals received BM-MNC injection with or without omentopexy. Controls received saline only. Three weeks after surgery, regional myocardial blood flow and contractility were measured, and density of arterioles was evaluated immunohistologically. Angiography and postmortem examinations were performed to determine collateral communication. RESULTS: Regional myocardial contractility was significantly improved by BM-MNC transplantation both with and without omentopexy (0.29 +/- 0.02 vs 0.11 +/- 0.03, p < 0.01, 0.30 +/- 0.02 vs 0.12 +/- 0.01, p < 0.01, respectively). Relative regional myocardial blood flow in the combined omentopexy group was significantly higher than the controls both at rest (1.05 +/- 0.11 vs 0.57 +/- 0.07, p < 0.01) and under stress (1.09 +/- 0.08 vs 0.40 +/- 0.10, p < 0.01). The number of arterioles (< 50 microm) in both groups were higher than the controls (88.1 +/- 5.00 vs 38.1 +/- 8.99, p < 0.01 and 109.2 +/- 9.91 vs 38.1 +/- 8.99, p < 0.01, respectively). The number of large arterioles (> 50 microm) in the combined omentopexy group was significantly higher than in both BM-MNC alone (26.9 +/- 2.4 vs 17.6 +/- 1.8, p = 0.011) and controls (26.9 +/- 2.4 vs 10.0 +/- 1.3, p < 0.01). Collateral communication between the omentum and myocardium was demonstrated by angiography and postmortem injection. CONCLUSIONS: The BM-MNC transplantation may attenuate cardiac contractile dysfunction, and omentopexy may enhance angiogenesis induced by BM MNC transplantation. PMID- 16368357 TI - Invited commentary. PMID- 16368358 TI - Novel measurement of relative aortic size predicts rupture of thoracic aortic aneurysms. AB - BACKGROUND: Optimal operative decision making in thoracic aortic aneurysms requires accurate information on the risk of complications during expectant management. Cumulative and yearly risks of rupture, dissection, and death before operative repair increase with increasing aortic size, but previous work has not addressed the impact of relative aortic size on complication rates. METHODS: Our institutional database contains data on 805 patients followed up serially with thoracic aortic aneurysms. Body surface area information was obtained on 410 patients (257 male, 153 female). We calculated a new measure of relative aortic size, the "aortic size index," and examined its ability to predict complications in these patients. RESULTS: Increasing aortic size index was a significant predictor of increasing rates of rupture (p = 0.0014) as well as the combined endpoint of rupture, death, or dissection (p < 0.0001). Using aortic size index, patients were stratified into three risk groups: less than 2.75 cm/m2 are at low risk (approximately 4% per year), 2.75 to 4.24 cm/m2 are at moderate risk (approximately 8% per year), and those above 4.25 cm/m2 are at high risk (approximately 20% per year). CONCLUSIONS: This study confirms that (1) thoracic aortic aneurysm is a lethal disease, (2) relative aortic size is more important than absolute aortic size in predicting complications, and (3) a novel measurement of relative aortic size allows for the stratification of patients into three levels of risk, enabling appropriate surgical decision-making. PMID- 16368359 TI - Surgical outcome of aortic arch repair for patients with Takayasu arteritis. AB - BACKGROUND: Takayasu arteritis can cause segmental dilatation or stenosis of the aorta and its major branches, and surgical treatment of it is still difficult. Our objective was to review late results of aortic arch repair for patients with Takayasu arteritis. METHODS: Between 1987 and 2003, 21 patients underwent aortic arch repair under circulatory arrest. Diagnosis was performed by pathologic study of specimens for all patients. Total aortic arch repair was performed in 12 patients with separated branched grafts and in 2 patients with the island technique. Selective cerebral perfusion was used in 12 patients and retrograde cerebral perfusion in 2 patients in this type of surgery. Hemiarch replacement using retrograde cerebral perfusion was performed in 7 patients. Craniocervical vascular stenosis was found in 7 patients and aneurysm in 5 patients. The elephant trunk technique was used in 10 patients. The follow-up period was 6.2 +/ 4.2 years. RESULTS: There was one hospital death due to renal failure, and two late deaths, both of which were sudden. Late in follow-up, a patient who had undergone hemiarch replacement 12 years previously required total aortic arch repair for dilatation of the distal arch. Three patients required thoracoabdominal aortic repair and one patient descending aortic repair for residual aortic dilatation late in follow-up. Postoperative spinal infarction occurred in one patient who underwent hemiarch replacement. CONCLUSIONS: Surgical and late outcomes of aortic arch repair under circulatory arrest appear favorable, though late dilatation of the residual aorta is a matter of concern. PMID- 16368360 TI - Hypertonic saline dextran improves outcome after hypothermic circulatory arrest: a study in a surviving porcine model. AB - BACKGROUND: Hypertonic saline dextran (HSD) has been shown to have neuroprotective properties. In the present study we have assessed its potential neuroprotective efficacy in the setting of hypothermic circulatory arrest in a surviving porcine model. METHODS: Twenty-four pigs were randomized to receive two 5-minute intravenous infusions (4 mL/kg) of either HSD (7.5 % saline, 6% dextran 70) or normal saline immediately after and 4 hours after a 75-minute period of hypothermic circulatory arrest at a brain temperature of 18 degrees C. RESULTS: The 7-day survival was 75% in the HSD group and 66% in the control group (p > 0.9). Brain total histopathologic score was lower in the HSD group (p = 0.01). Postoperative behavioral scores were higher in the HSD group on the second day after surgery (p = 0.03). Intracranial pressure was lower in the HSD group from 45 minutes to 8 hours after hypothermic circulatory arrest (p = 0.03). Cerebral perfusion pressure was higher in the HSD group from 45 minutes to 3 hours after hypothermic circulatory arrest (p = 0.06). Brain lactate concentration was lower in the HSD group when compared with controls (p = 0.05). Furthermore, brain glucose levels tended to be higher and brain lactate-pyruvate ratio and lactate glucose ratio were lower in the HSD group. Brain tissue oxygen partial pressures were somewhat higher in the HSD group (p = 0.08). CONCLUSIONS: The use of HSD in experimental hypothermic circulatory arrest is associated with significantly better neurologic recovery, better histopathology, lower intracranial pressure, higher cerebral perfusion pressure, and better preservation of brain metabolism. PMID- 16368361 TI - Invited commentary. PMID- 16368362 TI - Chest reexploration in the intensive care unit after cardiac surgery: a safe alternative to returning to the operating theater. AB - BACKGROUND: The aim of this study was to determine the outcome of patients who had chest reexploration in the intensive care unit (ICU) for bleeding or cardiovascular instability after heart surgery. METHODS: This was a retrospective analysis of medical records of patients who had a chest reexploration in the ICU for bleeding or possible cardiac tamponade over a 9-year period (1991 to 2000), at the Cardiothoracic Centre of Manchester Royal Infirmary, England. RESULTS: Between 1991 and 2000, 240 patients (3.4% of the total heart operations) who fitted the above criteria were identified. Two hundred and seven (86%) were reexplored for bleeding, 22 (9%) for possible tamponade, and 11 (5%) for both. Ninety-five percent were reexplored within 24 hours (median, 5 to 6 hours). Two hundred and twenty-six patients were found to have bleeding on reexploration. Of these, 125 (55%) were found to have focal bleeding, 74 (33%) diffuse bleeding, and 11 (5%) both. Two hundred and twelve (88%) had their chest closed, 25 (12%) packed, and 13 (10%) had further chest openings while in ICU. Sixteen (6.7%) of the patients died. Seven (2.9%) had sternal wound infection. For the survivors, ICU stay ranged from 1 to 60 days (median, 1) and their hospital stay ranged from 2 to 90 days (median, 8). CONCLUSIONS: Chest reexploration in ICU for bleeding or tamponade after heart surgery can be a safe alternative to return to the operating theater. PMID- 16368363 TI - Physicians assistants in cardiothoracic surgery: a 30-year experience in a university center. AB - BACKGROUND: The purpose of this study was to evaluate our 30 years of experience with the use of physician assistants (PAs) on a cardiothoracic surgery (CTS) service. METHODS: A retrospective review of the utilization of CTS PAs was performed at a university center from 1973 to 2003. RESULTS: The number of PAs has increased from 2 (1973) to 23 (2003), corresponding to the increased clinical service demands with a constant resident number for the past decade. Physician assistant employment has expanded from one hospital to five hospitals in our university system where CTS is performed. The CTS service has expanded from 400 total cases per year (1973) to 4,000 cases (2002). We have had a 50% retention rate with duration of employment from 6 months to 28 years (11 PAs with service more than than 15 years). The PA role has changed little, with duties varying from history and physical examination, conduit harvesting, insertion of invasive catheters and chest tubes, surgical first assisting, closure of the chest, and optional primary intensive care unit night-time in-house call. Salary currently ranges from 55,000 dollars to 100,000 dollars depending on length of service and overall merit. Job satisfaction for PAs employed longer than 12 months has remained high. CONCLUSIONS: The addition of PAs to our CTS university service has allowed us to resolve many problems of work assignment and coverage and enabled us to establish effective and efficient surgical teams without increasing the number of categorical CTS residents. PMID- 16368364 TI - Complementary alternative medical therapies for heart surgery patients: feasibility, safety, and impact. AB - BACKGROUND: Complementary therapies (touch, music) are used as successful adjuncts in treatment of pain in chronic conditions. Little is known about their effectiveness in care of heart surgery patients. Our objective is to evaluate feasibility, safety, and impact of a complementary alternative medical therapies package for heart surgery patients. METHODS: One hundred four patients undergoing open heart surgery were prospectively randomized to receive either complementary therapy (preoperative guided imagery training with gentle touch or light massage and postoperative music with gentle touch or light massage and guided imagery) or standard care. Heart rate, systolic and diastolic blood pressure, and pain and tension were measured preoperatively and as pre-tests and post-tests during the postoperative period. Complications were abstracted from the hospital record. RESULTS: Virtually all patients in the complementary therapy group (95%) and 86% in standard care completed the study. Heart rate and blood pressure patterns were similar. Decreases in heart rate and systolic blood pressure in the complementary therapies group were judged within the range of normal values. Complication rates were very low and occurred with similar frequency in both groups. Pretreatment and posttreatment pain and tension scores decreased significantly in the complementary alternative medical therapies group on postoperative days 1 (p < 0.01) and 2 (p < 0.038). CONCLUSIONS: The complementary medical therapies protocol was implemented with ease in a busy critical care setting and was acceptable to the vast majority of patients studied. Complementary medical therapy was not associated with safety concerns and appeared to reduce pain and tension during early recovery from open heart surgery. PMID- 16368365 TI - Invited commentary. PMID- 16368366 TI - Hypertensive pulmonary vascular disease in adults with secundum or sinus venosus atrial septal defect. AB - BACKGROUND: The development of hypertensive pulmonary vascular disease (HPVD) is considered a risk factor in the long-term course of patients with secundum atrial septal defects (ASD). The aim of this study was to assess the prevalence and histologic degree of HPVD and pulmonary hypertension in relation to preoperative clinical and hemodynamic data, intraoperative findings, and operative outcome in adults. METHODS: Lung biopsies of 75 patients, mean age 44 +/- 14 years (18-71 years), with secundum ASD or sinus venosus defect including ten patients with partial anomalous pulmonary venous return were analyzed in accordance with preoperative and intraoperative findings as well as operative outcome. Lung biopsy was performed at the time of defect closure and was classified according to Heath and Edwards. RESULTS: Structural changes of the pulmonary vasculature were found in 59% of patients; grade 3 and higher changes were present in 19%. There were no statistically significant relations between histologic findings and preoperative clinical and hemodynamic data, intraoperative findings, and operative outcome. The prevalence of moderate (32-50 mm Hg) and severe (> 50 mm Hg) systolic pulmonary hypertension was 27% and 17%, respectively. Increased systolic pulmonary arterial pressure was associated with increased pulmonary vascular resistance (p < 0.000) and patients' age (p = 0.001). Patients with a lower functional capacity had a higher prevalence of pulmonary hypertension (p = 0.011). CONCLUSIONS: The prevalence of HPVD and pulmonary hypertension in adult patients with secundum ASD or sinus venosus defect is considerable. Preoperative hemodynamic data do not predict the degree of HPVD in lung biopsy. Closure is generally advised to prevent increasing pulmonary arterial pressure and decreasing functional capacity over time. PMID- 16368367 TI - Invited commentary. PMID- 16368368 TI - Truncus arteriosus associated with interrupted aortic arch in 50 neonates: a Congenital Heart Surgeons Society study. AB - BACKGROUND: Patients with both interrupted aortic arch (IAA) and truncus arteriosus (TA) have worse outcomes than those with either lesion in isolation. We determined outcomes and associated factors in this rare group. METHODS: From 1987 to 1997, 50 (11%) of 472 neonates with IAA were identified with TA. Site of aortic arch interruption was distal to the left subclavian artery in 16% and between the left common carotid and subclavian artery in 84%. From the common arterial trunk, the pulmonary arteries arose from a main pulmonary trunk in 46%, common orifice in 22%, and separate orifices in 32%. At presentation, truncal valve stenosis was present in 12% and regurgitation in 22%. RESULTS: There were 34 deaths, with a single early hazard phase. Overall survival from admission was 44%, 39%, and 31% at 6 months, 1 year, and 10 years, respectively. One patient had primary cardiac transplantation and 4 died without any intervention. The IAA repair alone was performed in 7 patients, with single stage repair of both IAA and TA in 38 patients. Associated factors for overall time-related death include female gender (p < 0.001), type III TA (p < 0.001) and one institution (low-risk; p < 0.001). Results improved somewhat over time (p < 0.001). At 5 years after IAA repair only 28% were alive without arch repair intervention, and at 5 years after TA repair only 18% were alive without conduit reoperation. CONCLUSIONS: The combination of IAA and TA carries high early mortality, with high risk of reinterventions in survivors. One stage repair of both TA and IAA is the optimal management. PMID- 16368369 TI - Invited commentary. PMID- 16368370 TI - Caval division technique for sinus venosus atrial septal defect with partial anomalous pulmonary venous connection. AB - BACKGROUND: Repair of sinus venosus atrial septal defect (ASD) with high partial anomalous pulmonary venous connection (PAPVC) using an internal patch may be complicated by obstruction of the superior vena cava (SVC) or pulmonary veins, or both, and sinus node dysfunction. In cases in which the anomalous veins insert more than 2 cm above the cavoatrial junction, we have adopted the technique of caval division in which the SVC is divided and the proximal end is anastomosed to the right atrial appendage, and the distal SVC serves as a conduit for pulmonary venous drainage to the left atrium through the ASD. We retrospectively compare the results of the internal patch repair versus the Warden technique. METHODS: Between 1991 and 2004, 54 patients diagnosed with sinus venosus ASD and PAPVC have undergone repair at our institution. Mean age was 13.4 years (range, 1.5 to 58). Thirteen patients (24%) had high insertion of anomalous veins and underwent the Warden technique. Follow-up averages 4.3 years (range, 1 to 13). RESULTS: There were no early or late deaths. All patients remain in normal sinus rhythm. Twelve of the 13 patients with Warden procedure have had postoperative echocardiograms, and 11 of these patients showed no evidence of SVC or pulmonary venous obstruction. In 1 patient, symptomatic pulmonary venous obstruction developed and required revision of a contracted intra-atrial pericardial baffle. CONCLUSIONS: Caval division for treatment of high PAPVC appears to be safe and is associated with low morbidity and mortality. The Warden procedure is an effective surgical option for patients undergoing correction of high PAPVC. PMID- 16368371 TI - Consequences of delayed surgical closure of patent ductus arteriosus in very premature infants. AB - BACKGROUND: Surgical closure of ductus arteriosus is commonly indicated in premature newborns. The aim of this study was to assess short-term and mid-term effects of delayed surgical closure of the ductus arteriosus on respiratory and digestive outcome in extremely preterm infants. METHODS: We retrospectively studied 58 infants less than 28 weeks gestational age who underwent surgical closure of ductus arteriosus between January 1997 and December 2002. Nine infants with intrauterine growth restriction and major congenital malformation were excluded from the study. Criteria for surgical closure of ductus arteriosus were: (1) medical treatment failure (ie, indomethacin or ibuprofen) and (2) hemodynamically patent ductus arteriosus: systemic arterial pressure less than gestational age in mm Hg, heart failure, left atrial-aortic root ratio greater than 1.6, mean velocity in the left pulmonary artery greater than 0.6 m/s, and ductus arteriosus diameter greater than 3 mm. Infants were divided into two groups: (1) the early group who had surgery before 21 days of life (n = 31), and (2) the late group who had surgery after 21 days of life (n = 27). Preoperative and postoperative criteria were compared between the two groups (ie, gestational age, birth weight, hemodynamic, ventilatory, and echographic [left atrial-aortic root ratio, mean velocity in the left pulmonary artery] parameters). RESULTS: Preoperative gestational age and birth weight did not differ between the two groups. In the early group, gestational age was 26 weeks (range, 23 to 28 weeks and birth weight was 800 g (range, 630 to 1,240 g). In the late group, gestational age was 26 weeks (range, 24 to 28 weeks) and birth weight was 840 g (530 to 1,130 g). Hemodynamic, ventilatory, and echographic parameters were similar in both groups. Rate of bronchopulmonary dysplasia was similar in both groups. However, at 24 hours post surgery, median FiO2 was higher in the late group (28% [range, 21% to 65%]) than in early group (21% [range, 21% to 60%]) (p < 0.05). Furthermore, full oral feeding was acquired later in the late group (57 days of life [range, 30 to 136 days]) than in the early group (37 days of life [range, 27 to 84 days]) (p < 0.01), and body weight at 36 weeks of post conceptional age was higher in the early group at 1,800 g (range, 1,250 to 2,750 g) than in the late group at 1,607 g (1,274 to 2,200 g) (p < 0.05). CONCLUSIONS: Our findings show that early surgical closure of the ductus arteriosus (< 3 weeks of life) is associated with shortened delay for full oral feeding and improved body growth when compared with late surgical closure (> 3 weeks of life). PMID- 16368372 TI - Invited commentary. PMID- 16368373 TI - Closed transventricular aortic valvotomy for critical aortic stenosis in neonates: outcomes, risk factors, and reoperations. AB - BACKGROUND: Critical aortic stenosis (AS) in neonates necessitates urgent intervention for patient survival. The optimal treatment, however, continues to be controversial and still has high morbidity and mortality in many centers. This study examined our late outcome, risks, and reoperations after the treatment of critical AS in neonates. METHODS: Sixty-six neonates (47 boys and 19 girls) underwent closed transventricular aortic valvotomy for critical AS between 1978 and 2000. The mean age at the first intervention was 15.1 +/- 19.6 days (range, 1 to 78). Their weight ranged from 1.4 to 6.2 kg (mean, 3.5 +/- 0.9 kg). Sixteen patients (23%) had isolated critical AS and normal or dilated left ventricles, and 50 neonates (77%) had associated cardiovascular anomalies (ie, aortic annular hypoplasia, hypoplasia of the left ventricle, mitral valve abnormalities, and endocardial fibroelastosis, coarctation, or interruption of aorta). RESULTS: The hospital mortality was 29% (19 of 66). Kaplan-Meier 5- and 15-year survival was 61% and 58%, respectively. The operative mortality rates were 6% in neonates with isolated AS and 36% in patients with complex AS. The five risk factors significant for mortality on univariate analysis were (1) presence of endocardial fibroelastosis (p = 0.05), (2) presence of hypoplastic left ventricle (p = 0.003), (3) presence of associated cardiovascular anomalies (p = 0.04), (4) aortic valve annulus of less than 5.0 mm (p = 0.01), and (5) surgery before 1985 (p = 0.003). Of these five factors, only the presence of hypoplastic left ventricle (p = 0.001) and surgery before 1985 (p = 0.001) remain significant for mortality by multivariate analysis. At last follow-up (mean, 8.2 +/- 6.2 years), 36 of 47 of the long-term survivors were in New York Heart Association functional class I and II. Kaplan-Meier analysis showed 5- and 15-year freedom from aortic valve reoperation to be 83% and 60%, respectively. Univariate and multivariate analysis showed the presence of multilevel stenosis (p = 0.04) as the best preoperative predictor for the need for late reoperation in survivors. CONCLUSIONS: We conclude that critical AS in neonates continues to be associated with significant mortality. Reintervention such as surgical and balloon valvotomy are usually required within 10 years of initial surgery. PMID- 16368374 TI - Invited commentary. PMID- 16368375 TI - Muscle-sparing extrapleural approach for the repair of aortic coarctation. AB - BACKGROUND: This paper describes a muscle-sparing, extrapleural approach to repair aortic coarctation, and evaluates the results with established standards. METHODS: Forty consecutive patients with aortic coarctation (median age, 8 days; weight, 3.3 kg) were approached with a less invasive technique consisting of a short posterior thoracotomy, with only minimal (24 patients) or no (16 patients) division of thoracic wall muscles and a subperiosteal-extrapleural approach. Extended resection of the coarctation with enlargement of the distal aortic arch was performed in all patients. The median cross-clamp and operative times were 22 and 90 minutes, respectively. RESULTS: The repair was possible in all patients without needing conversion. There was no intraoperative or postoperative related complication. Two patients died early of low cardiac output as a result of ventricular fibroelastosis and respiratory failure. One patient died late of unrelated cause. The perioperative mean gradients across the neoarch were less than 5 mm Hg in all but 3 patients with proximal (2 patients) or mid arch (1) stenosis. The median ventilation time, intensive care unit stay, and hospital stay in isolated coarctation repairs was 2, 4.5, and 11 days, respectively. One patient had a recurrent stenosis at the site of surgical repair. Two patients underwent successful balloon dilatation, and 2 had surgical enlargement plasty of the proximal aortic arch at the time of intracardiac repair. None of the patients required chronic antihypertensive medication. At 29 months, freedom from reintervention on the isthmus and arch plus isthmus was 97.1% and 89.7%, respectively. CONCLUSIONS: A muscle-sparing, extrapleural approach for the repair of aortic coarctation is possible and provides results similar to conventional techniques. The approach reduces postoperative morbidity related to division of thoracic wall muscles and handling of the lung, restores a normal intercostal space, and produces superior cosmetic results, while at the same time leading to early and permanent relief of proximal hypertension. PMID- 16368376 TI - Maternal-fetal interactions in fetal cardiac surgery. AB - BACKGROUND: We examined potential maternal-fetal interactions during fetal cardiopulmonary bypass. these interactions, not previously described, may significantly influence attempts at fetal cardiac surgery. METHODS: Eight fetal sheep underwent cardiopulmonary bypass (5 singletons, 3 twins; 100-109 days) for 60 minutes using a centrifugal microcircuit (20 mL prime), and the placenta as oxygenator. We measured maternal hemodynamics, arterial blood gases, and changes in blood flow to the gravid uterus using bilateral uterine artery flow probes. Maternal measurements were correlated to fetal hemodynamics, blood gases, and umbilical blood flows. After bypass, fetuses were followed for 60 minutes. RESULTS: Decreases in uterine blood flow occurred without changes in maternal hemodynamics or arterial blood gases, but were associated with worsening fetal arterial blood gases (pH decreased from 7.2 +/- 0.2 to 7.0 +/- 0.1, partial pressure of carbon dioxide increased 45.6% and partial pressure of oxygen decreased 15.4%). Changes in maternal hemodynamics (decreased systolic blood pressure [17.5%, SD = 11] and decreased diastolic blood pressure [20.3%, SD = 15]) were only noted when uterine blood flows decreased by greater than 38.2% (SD = 26). Correction of maternal hypocalcemia (0.89 g/dL, SD = 0.1) led to improved uterine artery flows (28.3% increase, SD = 30). Finally, fetal sternotomy, cannulation, and cardiopulmonary bypass each decreased uterine artery flows by 27.5% (SD = 18), 31.0% (SD = 26), and 39.7% (SD = 25), respectively. Similar changes were not observed in the nonbypass twin. CONCLUSIONS: Significant changes in uterine blood flow can occur during fetal cardiopulmonary bypass support without apparent changes in maternal hemodynamics or arterial blood gases. These changes imply a unique transplacental maternal-fetal interaction. Limited data from the twin fetus suggest a localized mechanism involving only the segment of placenta exposed to extracorporeal circulation. PMID- 16368377 TI - Neonatal cavopulmonary assist: pulsatile versus steady-flow pulmonary perfusion. AB - BACKGROUND: Morbidity and mortality associated with single-ventricle physiology decrease substantially once a systemic venous, rather than systemic arterial, source of pulmonary blood flow is established. Cavopulmonary assist has potential to eliminate critical dependence on the problematic systemic-to-pulmonary shunt as a source of pulmonary blood flow in neonates. We have previously demonstrated feasibility of neonatal cavopulmonary assist under steady-flow conditions. We hypothesized that pulsatile pulmonary perfusion would further improve pulmonary hemodynamics. METHODS: Lambs (weight 7.2 +/- 1.1 kg, age 7.9 +/- 1.5 days) underwent total cavopulmonary diversion using bicaval venous-to-main pulmonary artery cannulation. A miniature centrifugal pump was used to augment cavopulmonary flow. Pulsatility was created with an intermittently compressed compliance chamber in the circuit. Hemodynamic and gas exchange data were measured for 8 hours. Pulsatile (n = 6), steady-flow (n = 13), and control (n = 6) groups were compared using two-way analysis of variance with repeated measures. RESULTS: All animals remained physiologically stable with normal gas exchange function. Mean pulmonary arterial pressure was elevated in pulsatile and steady-flow groups compared with the control group and within-group baseline values. Pulmonary vascular resistance was elevated initially in both assist groups but decreased significantly over the last 4 hours of the study and normalized after hour 4 in the pulsatile perfusion group. Pulmonary vascular resistance also normalized to control in the steady-flow group after hour 7. CONCLUSIONS: Both steady-flow and pulsatile pulmonary perfusion demonstrated normalization of pulmonary vascular resistance to control in a neonatal model of univentricular Fontan circulation. These results suggest that there is no benefit to pulsatile flow in this model. PMID- 16368378 TI - Twenty-four hour cardiopulmonary stability in a model of assisted newborn Fontan circulation. AB - BACKGROUND: Morbidity and mortality after stage-1 palliation of hypoplastic left heart syndrome is high as a result of adverse physiologic conditions imposed by the systemic-to-pulmonary arterial shunt. Conversion to a systemic venous source of pulmonary blood flow (Glenn/Fontan) substantially decreases instability and mortality risk. Cavopulmonary assist has the potential to eliminate critical dependence on the problematic systemic arterial shunt. We studied this support modality during a 24-hour period in a neonatal animal model of univentricular Fontan circulation. METHODS: Lambs (8.1 +/- 0.9 kg, 8.3 +/- 2.1 days, n = 7) underwent total cavopulmonary diversion. A miniature centrifugal pump was used to assist cavopulmonary flow. Control animals (6.6 +/- 1.0 kg, 7.3 +/- 2.1 days, n = 11) underwent placement of monitoring lines only. Hemodynamic and gas exchange data were measured. Within-group and between-group comparisons were made using two-way repeated measures analysis of variance. RESULTS: After an initial phase of reactivity, pulmonary vascular resistance returned to low levels and was not significantly different from baseline values after hour 13 or significantly different from control values after hour 4. Systemic venous pressure remained low. Oxygenation and ventilation remained normal with no histologic evidence of parenchymal lung injury. CONCLUSIONS: Pump-assisted cavopulmonary diversion is well tolerated up to 24 hours in the neonatal period. Despite initial reactivity, pulmonary vascular resistance trended toward normal and approached control values. Cavopulmonary assist holds the potential to serve as a bridge to neonatal Fontan repair of single ventricle. Chronic studies are warranted to determine the duration and rate of weaning of support to transition to an unassisted univentricular Fontan circulation. PMID- 16368379 TI - Differential effects of phosphodiesterase-5 inhibitors on hypoxic pulmonary vasoconstriction and pulmonary artery cytokine expression. AB - BACKGROUND: Perioperative pulmonary hypertension is a challenging clinical problem with numerous etiologies including hypoxia, adrenergic stimulation, and local inflammation. New oral phosphodiesterase-5 (PDE-5) inhibitors used for the treatment of erectile dysfunction may have beneficial effects on the pulmonary vasculature owing to the abundance of PDE-5 receptors in the lung. The purpose of this study was to compare the efficacy of sildenafil, vardenafil, and tadalafil in preventing acute hypoxic pulmonary vasoconstriction and hypoxia-induced pulmonary artery tumor necrosis factor-alpha (TNF-alpha) and interleukin-1-beta (IL-1beta) expression. METHODS: Isolated rat pulmonary arteries suspended in physiologic organ baths for measurement of isometric force transduction were treated with vehicle (dimethyl sulfoxide), sildenafil, vardenafil, or tadalafil to assess (1) pulmonary artery relaxation; (2) inhibition of phenylephrine induced pulmonary artery contraction; (3) inhibition of hypoxic pulmonary vasoconstriction (pO2 = 30-35 mm Hg); and (4) hypoxia-induced pulmonary artery TNF-alpha and IL-1beta expression (reverse transcriptase-polymerase chain reaction). RESULTS: Sildenafil, vardenafil, and tadalafil resulted in dose dependent pulmonary artery relaxation and inhibited phenylephrine-induced pulmonary artery contraction, but only tadalafil significantly inhibited hypoxic pulmonary vasoconstriction (52.08% +/- 7.65% tadalafil versus 88.63% +/- 8.96% vehicle; 98.61% +/- 10.04% sildenafil; 68.46% +/- 15.84% vardenafil). Hypoxia induced upregulation of TNF-alpha and IL-1beta mRNA in pulmonary artery was significantly decreased by tadalafil, but not sildenafil or vardenafil pretreatment. CONCLUSIONS: We conclude that sildenafil, vardenafil, and tadalafil were equally efficacious in causing pulmonary artery relaxation, but only tadalafil inhibited hypoxic pulmonary vasoconstriction and attenuated hypoxia induced pulmonary artery TNF-alpha and IL-1beta expression. PMID- 16368380 TI - Results of chest wall resection and reconstruction with and without rigid prosthesis. AB - BACKGROUND: Chest wall resections are associated with significant morbidity, with respiratory failure in as many as 27% of patients. We hypothesized that our selective use of a rigid prosthesis for reconstruction reduces respiratory complications. METHODS: The records of all patients undergoing chest wall resection and reconstruction were reviewed. Patient demographics, use of preoperative therapy, the location and size of the chest wall defect, performance of lung resection if any, the type of prosthesis, and postoperative complications were recorded. Predictor of complications were identified by chi2 and logistic regression analyses. RESULTS: From January 1, 1995, to July 1, 2003, 262 patients (median age, 60 years) underwent chest wall resection for tumor in 251 (96%), radiation necrosis in 7 (2.7%); and infection in 4 patients (1.3%). The median defect size was 80 cm2 (range, 2.7 to 1,200 cm2) and the median number of ribs resected was 3 (range, 1 to 8). Major lung resection was performed in 85 patients (34%). Prosthetic reconstruction was rigid (polypropylene mesh/methylmethacrylate composite) in 112 (42.7%), nonrigid (polytetrafluoroethylene or polypropylene mesh) in 97 (37%), and none in 53 patients. Postoperatively, 10 patients died (3.8%), 4 of whom had pneumonectomy plus chest wall resection. Respiratory failure occurred in 8 patients (3.1%). By multivariate analysis, the size of the chest wall defect was the most significant predictor of complications. CONCLUSIONS: Our incidence of respiratory failure is lower than previously reported and may relate to our use of rigid repair for defects likely to cause a flail segment. Pneumonectomy plus chest wall resection should be performed only in highly selected patients. PMID- 16368381 TI - Invited commentary. PMID- 16368382 TI - Are heart-lung transplant recipients protected from developing bronchiolitis obliterans syndrome? AB - BACKGROUND: Heart-lung transplant recipients, when compared with heart transplant recipients, are relatively spared from allograft coronary artery disease. This study was undertaken to investigate whether heart-lung transplant recipients are also spared from experiencing bronchiolitis obliterans syndrome (BOS) when compared with double-lung transplant recipients. In addition, the risk factors for developing BOS after lung transplantation were analyzed. METHODS: Heart-lung and bilateral sequential double-lung transplant recipients were reviewed retrospectively from 1990 to 2000 using the Stanford Transplant Database. The heart-lung transplant group consisted of 77 heart-lung transplant recipients and the double-lung transplant group consisted of 51 double-lung transplant recipients. The rates of BOS, survival, acute rejection, and cytomegalovirus infection at 1, 3, and 5 years were measured. RESULTS: There were no significant differences in patient demographics between the two groups. Rates of survival and acute rejection were similar in the two transplant groups. The incidence of cytomegalovirus infection was significantly higher in heart-lung transplant recipients. Freedom from BOS was similar in the two transplant groups. Risk factors for the development of BOS in the heart-lung and double-lung transplant recipients included male donor, younger recipient age, a diagnosis other than cystic fibrosis, nonuse of cardiopulmonary bypass, and the use of OKT3 induction therapy. CONCLUSIONS: Heart-lung transplant recipients exhibit BOS at a rate similar to double-lung transplant recipients. The immunoprotective effect the lung allograft presumably provides the heart is not reciprocated by the heart in preventing the development of BOS. PMID- 16368383 TI - Prognostic significance of metastasis to the highest mediastinal lymph node in nonsmall cell lung cancer. AB - BACKGROUND: We have tried to clarify the prognostic significance of metastasis to the highest mediastinal (HM) lymph node in patients with N2 lung cancer who underwent complete dissection of superior mediastinal (including HM) lymph nodes. METHODS: This study analyzed 53 patients with N2 nonsmall cell lung cancer who underwent surgical procedures such as lobectomy plus hilar and mediastinal node dissection (T4, neoadjuvant therapy cases were excluded). For patients whose cancer was in the left lung, we performed surgery through the median sternotomy in order to dissect superior mediastinal nodes. The clinicopathologic records of the patients were examined for prognostic factors such as age, sex, side, histology, tumor location, tumor size, clinical node (cN) number, preoperative serum carcinoembryonic antigen level, number of metastatic stations, and HM lymph node involvement. RESULTS: A univariate analysis showed that tumor size (T1/T2 3), cN factor (cN1-2/cN0), N2 level (multiple/single), and metastasis to the HM node were significant prognostic factors. In the multivariate analysis, metastasis to the HM lymph node remained a significant prognostic factor (p = 0.026). The 3-year survival rates were 52% in patients without metastasis to the HM lymph node and 21% in patients with metastasis to the HM lymph node (p < 0.001). Furthermore, when HM nodal involvement was absent, the 5-year survival rate was 33% even in patients with multilevel N2 status, 45% in patients with cN1 2 status, and 47% in patients with pT2-3 tumor status. CONCLUSIONS: Highest mediastinal lymph node involvement is prognostic of highly advanced N2 disease resulting in poor outcome. The results also suggest that patients with no involvement of the HM lymph node can experience acceptable postoperative outcomes even if they have multilevel N2 status, positive cN status, or T2-3 tumor status. PMID- 16368384 TI - Invited commentary. PMID- 16368385 TI - Pleural lavage cytology before and after lung resection in non-small cell lung cancer patients. AB - BACKGROUND: The aim of this study was to analyze on a multivariate basis the prognostic significance of pre-resection and post-resection pleural lavage cytologies in surgically resected primary non-small cell lung cancer (NSCLC) patients, in relation to pathologic TNM factors in a large cohort of almost 1,200 patients. METHODS: From August 1992 through March 2001, pleural lavage cytology (PLC) was performed in 1,214 NSCLC patients without pleural effusion or dissemination undergoing pulmonary resection. The cytologic evaluation was classified into three categories: negative, suggestive, and positive. To investigate the impact on patient survival, PLC results were analyzed with conventional clinicopathologic factors. RESULTS: Definitive pre-resection PLC result was obtained in 1,194 patients and 38 had a positive result. The 5-year survival rates were 27% if pre-resection PLC was positive and 71% if negative. Of 1,198 patients 54 had a positive post-resection PLC result. The 5-year survival rates were 10% if post-resection PLC was positive and 73% if negative. On multivariate analysis, post-resection PLC was an independent prognostic factor as significant as established clinicopathologic factors. CONCLUSIONS: Pre-resection and post-resection PLC should be recognized as an essential prognostic factor and should be performed in NSCLC patients without pleural effusion and dissemination. Post-PLC, compared with pre-PLC, had a greater and independent impact on survival and needs to be incorporated in the pathologic staging of NSCLC in the future. PMID- 16368386 TI - Invited commentary. PMID- 16368387 TI - Transsternal thymectomy for myasthenia gravis: surgical outcome. AB - BACKGROUND: Transsternal thymectomy is well established in the treatment of myasthenia gravis. Surgical strategy and patient selection, however, remain controversial. This paper reports the experience of a supraregional center looking into the influence of different preoperative risk factors on surgical outcome. METHODS: Between 1987 and 1998, 85 consecutive patients (65 female; mean age, 30.5 years) were enrolled. The mean preoperative Myasthenia Gravis Foundation of America stage was 2.3. The preoperative, early, and late follow-up data were analyzed retrospectively. RESULTS: Mean follow-up was 4.5 years (range, 1 to 14; 376 follow-up years). Mean duration of disease before surgery was 31 months. There were no operative or late deaths. Eight patients had major complications. Seventy-two patients were free from any early or late morbidity. Immunosupression therapy patients were more prone to have complications. At their last visit, 15 patients (17%) were in complete remission; 67 reported clinical improvement. Sixty-three were asymptomatic or in stage I on no or minimal treatment. Remission and clinical improvement were not predicted by patient's age, sex, duration of disease prior to surgery, thymic pathology, or antiacetylcholine receptor antibodies titer. Greater severity of symptoms before surgery was associated with greater subsequent improvement. Remission at 1 year predicted remission at the end of follow-up. CONCLUSIONS: Transsternal thymectomy for myasthenia gravis is safe and effective. It benefits most patients, especially those with severe symptoms. The long interval from diagnosis to surgery demonstrates it is never too late for thymectomy. PMID- 16368388 TI - Video-assisted thoracic surgery for pleural empyema. AB - BACKGROUND: After we gained considerable experience with video-assisted thoracic surgery (VATS) and became familiar with its advantages, we started to use it for the treatment of thoracic empyema. METHODS: We treated 130 patients with pleural empyema in whom chest tube drainage and antibiotic therapy had failed to produce a satisfactory result. Six months after surgery they had clinical and radiologic assessment and spirometry. RESULTS: Video-assisted surgery was performed in all patients. Mean operative time was 93 minutes (range, from 55 to 180 minutes), mean duration of postoperative chest tube drainage was 10 days (range, from 5 to 32 days), and mean hospital stay was 16 days (range, from 3 to 56 days). The rate of conversion to open thoracotomy was 3.1%. Complications for which reoperation was necessary occurred in 9% of patients. At follow-up after six months, the mean forced expiratory volume in 1 second was 87.7% (range, from 69.5% to 105.9%), the mean postoperative vital capacity was 84.4%, (range, from 59.9% to 97.9%). There were no postoperative or procedure-related deaths. CONCLUSIONS: Video-assisted thoracic surgery is a safe and effective treatment option for fibropurulent empyema with low morbidity and mortality. Conversion to thoracotomy should be used if necessary to remove all of the fibropurulent material and achieve complete expansion of the lung to insure a good outcome. PMID- 16368389 TI - A randomized clinical trial of lung volume reduction surgery versus best medical care for patients with advanced emphysema: a two-year study from Canada. AB - BACKGROUND: We present a summary report evaluating the efficacy of lung volume reduction surgery (LVRS) in patients with advanced emphysema in the Canadian setting. METHODS: Quality of Life measures assessed the efficacy of adding LVRS to best medical care including rehabilitation in this blinded randomized multicentered controlled trial with 2 years of follow-up. Health utility and quality-adjusted life years (QALY) were outcomes central to our economic assessment. RESULTS: None of the 32 patients randomized to the LVRS arm or 30 patients in the best medical care (BMC) arm crossed-over and no patients were lost to follow-up. Overall surgical mortality was 16% at 2 years while the overall medical mortality was 13% (p = 0.914). There were no 30-day postoperative deaths but 2 deaths (6%) occurred within 90 days of randomization. Surgery reduced the residual volume measured at 6 months by 23% (5,385 mL to 4,322 mL, p = 0.007). There was an increase in forced expiratory volume in 1 second (FEV1) of 30% (265 mL, p = 0.013) from baseline, an improvement in the six minute walk test (6MWT) of 78 meters (p = 0.045), and an increase in Health Utility Index 3 (HUI3) which peaked at 6 months with a difference of 0.16 (p = 0.129). There was a gain in QALYs of 0.21 (p = 0.19) in the LVRS-arm over the BMC-arm. The LVRS costs an additional 28,119 Canadian dollars (CAD) compared with BMC or 133,900 CAD/QALY gained. CONCLUSIONS: The addition of LVRS to best medical care including pulmonary rehabilitation improves pulmonary function, exercise activity, and quality of life in selected patients with advanced emphysema. Cost is high but in keeping with other treatment modalities currently available. PMID- 16368390 TI - Ultrasound mini-transducer with malleable handle for coronary artery surgery. AB - PURPOSE: A 13 MHz epicardial ultrasound mini-transducer (15 x 9 x 6 mm) with a custom made malleable handling tool was evaluated to assess the coronary artery and anastomosis on all sides of the heart. DESCRIPTION: On indication, in 8 patients undergoing coronary artery bypass surgery on the arrested heart, 8 coronary arteries as well as 27 coronary anastomoses were scanned. EVALUATION: The malleable handle was easily adjusted, and all sides of the heart were accessible for scanning with the mini-transducer. Based on intraoperative scanning, the anastomotic site was altered (n = 4), an additional coronary artery was grafted (n = 2), and the left anterior descending coronary artery was identified after incorrect conventional selection of the diagonal branch (n = 1). No anastomosis construction errors were detected. In one anastomosis, a calcified plaque was seen in the outflow corner. CONCLUSIONS: The epicardial ultrasound mini-transducer with its malleable handle allowed successful visualization and assessment of the coronary arteries and anastomoses on all sides of the heart. Ultrasound information greatly aided in intraoperative decision making that resulted in anastomotic site changes and prevented grafting of the wrong vessel. PMID- 16368391 TI - Invited commentary. PMID- 16368392 TI - Intraoperative sentinel lymph node mapping using a new sterilizable magnetometer in patients with nonsmall cell lung cancer. AB - PURPOSE: We recently developed a novel method for sentinel lymph node mapping using magnetic force. However, problems with the sterility and sensitivity of the magnetometer made intraoperative sentinel lymph node mapping impossible. The purpose of this study was to test the utility of a new, more sensitive, sterilizable magnetometer developed in our institute for in vivo sentinel lymph node mapping in patients with nonsmall cell lung cancer. DESCRIPTION: Ferumoxides (magnetite) served as the tracer. Twenty patients with nonsmall cell lung cancer participated in the study. Each received 5 mL of ferumoxides, which were injected around their tumor. Magnetic force was then measured intraoperatively using the new sterilizable magnetometer. EVALUATION: The in vivo sentinel lymph node detection rate was 80.0% (16 of 20). The accuracy, sensitivity, and false negative rates were 100% (16 of 16), 100% (4 of 4) and 0% (0 of 12), respectively. Our preliminary study indicates that our new magnetometer enables in vivo sentinel lymph node mapping in patients with nonsmall cell lung cancer. CONCLUSIONS: This device safely and accurately detected sentinel lymph nodes in nonsmall cell lung cancer patients. PMID- 16368393 TI - Invited commentary. PMID- 16368394 TI - The use of flexible silastic drains after chest surgery: novel thoracic drainage. AB - PURPOSE: We report a new strategy for drainage with silicon thoracic tube (Blake drain) after chest surgery. DESCRIPTION: To confirm the effect of Blake drain, we have performed a three-part study including in vitro and clinical investigations compared with those of conventional chest tubes. We carried out an in vitro analysis to achieve the best possible drainage; in the second part, we used this drain in a cohort of 30 patients to establish safety and efficacy; and in a third substudy, we carried out a nonrandomized comparison with an earlier cohort between the Blake drain group and standard, rigid drain group. EVALUATION: In vitro tests demonstrate that the drainage capability of the Blake drain depends on sufficient length in the fluted part of the structure. Clinical outcome demonstrates no significant differences. The Silastic drain (Ethicon, Inc, Somerville, NJ) group had a significantly shorter period of tube drainage compared with the conventional drain group. CONCLUSIONS: From this small study the Blake drains seem to be safe and effective. Therefore, a prospective, randomized comparison should be carried out. PMID- 16368395 TI - Invited commentary. PMID- 16368396 TI - A piece of glass in the heart. AB - Most posttraumatic foreign bodies are metal objects in the heart, such as bullets and needles; nonmetal objects are very rare. We now report a case in which a piece of glass entered a girl's heart from an injury to the right side of her neck. Six months later, we successfully removed the glass shard from the anterior wall of the right ventricle. PMID- 16368397 TI - Heart surgery in infants with hemophilia. AB - We describe 2 infants with hemophilia A who had heart surgery under cardiopulmonary bypass with factor VIII replacement therapy, and we recommend a guideline for factor VIII support for cardiac surgery. One child had repair of total anomalous pulmonary venous connection. The second had cardiac catheterization followed by repair of ventricular septal defect and total anomalous pulmonary venous connection. Close collaboration between hematologist, laboratory, cardiologist, and cardiac surgeon is crucial in successful management of coagulation abnormalities during and after surgery. PMID- 16368398 TI - Infectious ductal aneurysm after coil embolization in an infant. AB - We report a case of ductal aneurysm after transcatheter coil embolization in an infant. The aneurysm was asymptomatic and was not detected until it ruptured during surgical intervention. We suspect that the aneurysm was induced by methicillin-resistant Staphylococcus aureus infection, as the resected aneurysmal wall was severely infiltrated by inflammatory cells and the patient had recurrent methicillin-resistant Staphylococcus aureus infection. We recommend remaining on guard against formation of a ductal aneurysm after coil embolization, especially in patients with associated recurrent bacteremia. PMID- 16368399 TI - Hypoplastic left heart syndrome with anomalous origin of the right coronary artery. AB - Hypoplastic left heart syndrome in association with an anomalous origin of a coronary artery from the pulmonary artery is a very rare congenital malformation. In the few reported cases, the left coronary artery or the circumflex artery arises from the right pulmonary artery. We describe a newborn who presented with hypoplastic left heart syndrome, and at the time of operation had an anomalous origin of the right coronary artery from the right pulmonary artery that was detected. The patient underwent a successful modified Norwood procedure with direct reimplantation of the right coronary artery to the neo-aorta. PMID- 16368400 TI - Anterior approach to traumatic mid aortic arch transection. AB - The majority of nonpenetrating traumatic injuries to the thoracic aorta are fatal. Survivors of aortic transection tend to have injuries occurring at the isthmus. We report a rare, blunt traumatic complete transection of the mid aortic arch between the innominate and left common carotid arteries diagnosed by multidetector computed tomography of the chest. The repair was approached anteriorly and required aortic arch replacement. PMID- 16368401 TI - Successful repair of iatrogenic acute aortic dissection with cerebral malperfusion. AB - We describe the successful treatment of a patient with iatrogenic acute aortic dissection including cerebral malperfusion as a complication of coronary artery surgery. After beginning cardiopulmonary bypass, a retrograde ascending aortic dissection associated with cerebral malperfusion was recognized. Systemic circulation was immediately arrested at 31 degrees C. After aortotomy, hypothermic selective antegrade cerebral perfusion was established. Replacement of the ascending aorta with coronary artery bypass grafting was performed without neurologic complications. PMID- 16368402 TI - Traumatic partial avulsion of a single right subclavian artery from the aortic arch and definitive repair. AB - Blunt injury to the right subclavian artery is a rare complication of severe deceleration trauma often associated with significant morbidity and mortality. We describe an atypical presentation in a patient who sustained a traumatic avulsion of his right subclavian artery arising off the aortic arch. An interposition graft was used to restore the continuity of the artery to the ascending thoracic aorta. PMID- 16368403 TI - Novel repair for late posttraumatic aortic valve injury and root pseudoaneurysm. AB - We report a case of posttraumatic aortic valve regurgitation and pseudoaneurysm of the aortic root diagnosed 22 months after a road traffic accident. The surgical treatment consisted of exclusion of the pseudoaneurysm with direct closure of the entry tear in the right coronary sinus followed by insertion of a Toronto stentless prosthesis (St. Jude Medical, St. Paul, MN). This surgical approach aimed to cover the repaired entry tear ensuring exclusion of the site. Recovery was uneventful and 12 month follow-up was unremarkable. PMID- 16368404 TI - Technical considerations for myocardial revascularization in congenital bilateral hypoplasia of the pericardium. AB - Congenital hypoplasia of the pericardium is a rare clinical condition that is typically encountered as an incidental finding during routine thoracic imaging or cardiothoracic surgery. Chest pain symptoms, when they occur, are often initially attributed to coronary ischemic syndromes, but herniation of cardiac structures through the pericardial defect, ischemia from torsion of great vessels, or compression of pulmonary parenchyma through defects between the aortic and pulmonary arteries can occur. Careful attention to cannulation techniques, conduit length, and the aberrant course of the phrenic nerves, typically through pericardial fibrous bands, is critical to the successful conduct of cardiac surgery. PMID- 16368405 TI - Dysphagia in the young male: the ringed esophagus. AB - The endoscopic view of the multi-ringed esophagus readily explains why the term "tracheal esophagus" is applicable. This entity may be undiagnosed until dysphagia and impactions secondary to strictures occur in the young male. Several factors point to a congenital rather than an acquired disorder. Treatment consists of slow, progressive dilatations that are repeated for recurrent dysphagia. PMID- 16368407 TI - Bleeding "downhill" varices: a rare complication of intrathoracic goiter. AB - We report a case of a 68-year-old woman with a 16-year history of a benign goiter. The patient presented with hematemesis and a clinically evident superior vena cava syndrome. Preoperative evaluation demonstrated upper esophageal varices and large posterior mediastinal goiter. The patient underwent successful resection of the mass with complete resolution of both the superior vena cava syndrome and "downhill" varices. PMID- 16368406 TI - Gastropericardial fistula, purulent pericarditis, and cardiac tamponade after laparoscopic Nissen fundoplication. AB - Gastropericardial fistula, purulent pericarditis, and cardiac tamponade developed 7 years after laparoscopic Nissen fundoplication. The patient was successfully managed through a thoracotomy by open drainage of the pericardium, excision and closure of the fistula, and an omentum flap. PMID- 16368408 TI - Pulmonary artery dissection complicating lung transplantation for primary pulmonary hypertension. AB - We report a case of pulmonary artery (PA) dissection complicating lung transplantation. A 30-year-old man with primary pulmonary hypertension underwent bilateral single lung transplantation. Thirty hours postoperatively he had signs of obstruction of the right main pulmonary artery develop due to dissection demonstrated on transesophageal echocardiography and pulmonary arteriography. Surgical repair of the dissection with reanastomosis of the pulmonary artery was successfully carried out, and the patient subsequently improved to a favorable condition. Pulmonary hypertension is a risk factor of pulmonary artery dissection; our case suggested that underlying chronic pulmonary hypertension might have led to this rare and life-threatening anastomotic problem after lung transplantation. PMID- 16368409 TI - Presentation and management of a stage Ia lung cancer patient with a paraneoplastic factor VIII inhibitor. AB - Acquired inhibitors to clotting factors most commonly involve factor VIII and are associated with autoimmune disease. Factor VIII inhibitors can cause severe spontaneous and iatrogenic bleeding that is difficult to manage. Factor VIII inhibitors are rarely associated with solid tumors and only three cases of adenocarcinoma of the lung have been reported. This report describes the multidisciplinary management of a factor VIII inhibitor-producing stage Ia lung adenocarcinoma that ultimately resulted in complete resectability. PMID- 16368410 TI - Bronchopleural fistula treated with a covered wallstent. AB - Bronchopleural fistula is a well-recognized complication of pneumonectomy, which presents a difficult challenge to the thoracic surgeon. We report the successful treatment of a bronchopleural fistula after right pneumonectomy for lung cancer, using a covered esophageal stent. PMID- 16368411 TI - ACTH-secreting thymic carcinoid associated with multiple endocrine neoplasia type 1. AB - Thymic carcinoids are classified into three categories: (1) nonsecretory tumors, (2) hormonal secretory tumors, and (3) tumors associated with multiple endocrine neoplasia type 1. We report a rare case with adrenocorticotropic hormone secreting thymic carcinoid with multiple endocrine neoplasia type 1. Radiologic examination showed an anterior mediastinal mass and a parathyroid tumor. Blood analysis revealed high levels of parathyroid hormone and adrenocorticotropic hormone. Urine cortisol and 17-hydroxycorticoids levels were also elevated. Extended thymectomy was performed. Subsequently adjuvant radiation therapy and parathyroid tumor resection were performed. A germline mutation of exon 7 in the multiple endocrine neoplasia type 1 gene was detected and a somatic mutation of exon 9 was demonstrated in the thymic tumor. PMID- 16368412 TI - Supradiaphragmatic bronchogenic cyst extending into the retroperitoneum. AB - We experienced a case of bronchogenic cyst located on the left lower pleural space extending into the retroperitoneum in a 33-year-old man with left chest pain. Preoperative imaging studies and operative findings showed a broad-based cystic mass on the left lower posterior pleura and diaphragmatic pleura extending into the retroperitoneal region across the diaphragm. Histologic studies proved this mass to be a bronchogenic cyst. PMID- 16368413 TI - A life-threatening complication of the Nuss procedure for pectus excavatum. AB - We describe a delayed, life-threatening complication in a boy operated on using the Nuss-procedure 2 months earlier. On admittance he was in shock with cardiac tamponade. An immediate needle aspiration of blood from the pericardium was done before a median sternotomy. The Nuss-bar was removed, and we identified a 1.5-cm laceration in the adventitial layer of the ascending aorta as the source of bleeding. The tear was closed, and the patient had an uneventful recovery. Careful positioning of the bar is necessary to avoid complications. Measures must be taken postoperatively to confirm that the steel bar does not rotate. PMID- 16368414 TI - Magnetic resonance images of diaphragmatic endometriosis treated by polyglactin mesh. PMID- 16368415 TI - Intrapericardial teratoma presenting with circulatory compromise. PMID- 16368416 TI - Choice of artificial chordae length according to echocardiographic criteria. AB - A simple way to identify artificial chordae length is reported. The distance A between the edge of the prolapsing anterior leaflet and the plane of the mitral annulus is measured during perioperative transesophageal echocardiography. When the mitral valve is exposed, the elongated chorda, corresponding to the scallop previously evaluated, is measured with a ruler and A is subtracted. This is the length of the new chorda that is measured with a ruler and tied at that level after being properly positioned. PMID- 16368417 TI - Novel technique for volume reduction of giant left atrium: simple and effective "spiral resection" method. AB - Despite some controversial studies, an enlarged left atrium has an impact on postoperative complications, and surgical correction should be considered, particularly in patients with a markedly enlarged left atrium. We present herein 2 patients with a giant left atrium (left atrial dimensions of 107 and 93 mm on echocardiograms), and describe an effective and simple procedure, the "spiral resection" method, to reduce any part of the dilated wall of the left atrium with a single incision. PMID- 16368418 TI - Alternative technique of the right atrial anastomosis (cavo-atrial) in orthotopic heart transplantation. AB - An alternative technique of the right atrial anastomosis in heart transplantation, which allows a more anatomical reconstruction of the right atrium and is easier in handling than bi-caval anastomoses is described. PMID- 16368419 TI - Wrapping of the left internal thoracic artery with an expanded polytetrafluoroethylene membrane. AB - We describe the wrapping of the proximal segment of the left internal thoracic artery graft in a polytetrafluoroethylene membrane. Two groups of patients were compared (99 patients with wrapping, 70 patients as controls). There were no statistical differences between the two groups regarding the postoperative course. Three patients in the polytetrafluoroethylene group and 2 in the control group underwent reoperation for valve surgery. Exposure of the wrapped graft segment for clamping was safer and more rapid than in the control group. PMID- 16368420 TI - Myocardial thievery: the coronary-subclavian steal syndrome. AB - Coronary-subclavian steal syndrome entails the reversal of blood flow in a previously constructed internal mammary artery coronary conduit, which produces myocardial ischemia. The most frequent cause of the syndrome is atherosclerotic disease in the ipsilateral, proximal subclavian artery. Although coronary subclavian steal was initially reported to be rare, the increasing documentation of this phenomenon and its potentially catastrophic consequences in recent series suggests that the incidence of the problem has been underreported and that its clinical impact has been underestimated. We review the causes and background of coronary-subclavian steal; methods of preventing, diagnosing, and treating it; and the potential influence of various treatment regimens on long-term survival and the likelihood of late adverse events in patients with coronary-subclavian steal syndrome. PMID- 16368421 TI - Large pedunculated polyps originating in the esophagus and hypopharynx. AB - Large pedunculated esophageal and hypopharyngeal polyps are uncommon. Clinical presentation most commonly includes dysphagia and mass regurgitation. If left untreated the patient may develop polyp aspiration complicated by fatal asphyxiation. Diagnosis has depended on endoscopy and barium swallow historically; however, these procedures may fail to provide a diagnosis. In recent years computerized tomographic scan and magnetic resonance imaging have proved reliable methods of diagnosis. These polyps are located predominantly in the upper esophagus and frequently are comprised of a fibrous component. Malignant potential is low. Resection of these lesions is warranted; it may be approached endoscopically if feasible or surgically through a cervical or thoracotomy approach, depending on the location. Recurrence is rare. To our knowledge, this is the largest review of large esophageal polyps, including 110 reported cases in the literature. PMID- 16368422 TI - The Society of Thoracic Surgeons Practice Guideline Series: Antibiotic Prophylaxis in Cardiac Surgery, Part I: Duration. PMID- 16368423 TI - Is tricuspid valve annuloplasty required in children with right heart obstruction and tricuspid valve regurgitation? PMID- 16368424 TI - Skeletonization of the internal thoracic artery: pros and cons. PMID- 16368426 TI - The signal-averaged P-wave to predict atrial fibrillation after cardiac surgery. PMID- 16368427 TI - Minimally invasive surgical placement of left ventricular epicardial lead: letter 2. PMID- 16368428 TI - Minimally invasive surgical placement of left ventricular epicardial lead: letter 1. PMID- 16368429 TI - Digital recording of operations. PMID- 16368431 TI - Prognosis of lung cancer in heart transplant recipient. PMID- 16368432 TI - Galectin-8 binds specific beta1 integrins and induces polarized spreading highlighted by asymmetric lamellipodia in Jurkat T cells. AB - Integrin-mediated encounters of T cells with extracellular cues lead these cells to adhere to a variety of substrates and acquire a spread phenotype needed for their tissue incursions. We studied the effects of galectin-8 (Gal-8), a beta galactoside binding lectin, on Jurkat T cells. Immobilized Gal-8 bound alpha1beta1, alpha3beta1 and alpha5beta1 but not alpha2beta1 and alpha4beta1 and adhered these cells with similar kinetics to immobilized fibronectin (FN). Function-blocking experiments with monoclonal anti-integrin antibodies suggested that alpha5beta1 is the main mediator of cell adhesion to this lectin. Gal-8, but not FN, induced extensive cell spreading frequently leading to a polarized phenotype characterized by an asymmetric lamellipodial protrusion. These morphological changes involved actin cytoskeletal rearrangements controlled by PI3K, Rac-1 and ERK1/2 activity. Gal-8-induced Rac-1 activation and binding to alpha1 and alpha5 integrins have not been described in any other cellular system. Strikingly, Gal-8 was also a strong stimulus on Jurkat cells in suspension, triggering ERK1/2 activation that in most adherent cells is instead dependent on cell attachment. In addition, we found that patients with systemic lupus erythematosus (SLE), a prototypic autoimmune disorder, produce Gal-8 autoantibodies that impede both its binding to integrins and cell adhesion. These are the first function-blocking autoantibodies reported for a member of the galectin family. These results indicate that Gal-8 constitutes a novel extracellular stimulus for T cells, able to bind specific beta1 integrins and to trigger signaling pathways conducive to cell spreading. Gal-8 could modulate a wide range of T cell-driven immune processes that eventually become altered in autoimmune disorders. PMID- 16368433 TI - Deficient E-cadherin adhesion in C57BL/6J-Min/+ mice is associated with increased tyrosine kinase activity and RhoA-dependent actomyosin contractility. AB - The Min/+ mouse is a model for APC-dependent colorectal cancer (CRC). We showed that tumorigenesis in this animal was associated with decreased E-cadherin adhesion and increased epidermal growth factor receptor (Egfr) activity in the non-tumor intestinal mucosa. Here, we tested whether these abnormalities correlated with changes in the actin cytoskeleton due to increased Rho-ROCK signaling. We treated Apc+/+ (WT) littermate small intestine with EGTA, an inhibitor of E-cadherin, and with LPA, an RhoA activator; both induced effects on adhesion and kinase activity that mimicked the Min/+ phenotype. GTP-bound Rho was increased in Min/+ enterocytes relative to WT. Since RhoA activity is associated with actomyosin contractility, markers of this signaling cascade were assessed including phosphorylated myosin light chain (MLC), cofilin, Pyk2, Src, and MAPK kinases. The increased actomyosin contractility characterizing Min/+ intestinal tissue was suppressed by the ROCK inhibitor, Y27632, but was inducible in the WT by EGTA or LPA. Finally, ultrastructural imaging revealed changes consistent with actomyosin contractility in Min/+ enterocytes. Thus, the positive regulation of E cadherin adhesion provided by Apc+ in vivo allows proper negative regulation of Egfr, Src, Pyk2, and MAPK, as well as RhoA activities. PMID- 16368434 TI - Analysis of the influence of subcellular localization of the HIV Rev protein on Rev-dependent gene expression by multi-fluorescence live-cell imaging. AB - The human immunodeficiency virus Rev protein is a post-transcriptional activator of HIV gene expression. Rev is a nucleocytoplasmic shuttle protein that displays characteristic nuclear/nucleolar subcellular localization in various cell lines. Cytoplasmic localization of Rev occurs under various conditions disrupting Rev function. The goal of this study was to investigate the relationship between localization of Rev and its functional activity in living cells. A triple fluorescent imaging assay, called AQ-FIND, was established for automatic quantitative evaluation of nucleocytoplasmic distribution of fluorescently tagged proteins. This assay was used to screen 500 rev genes generated by error-prone PCR for Rev mutants with different localization phenotypes. Activities of the Rev mutants were determined with a second quantitative, dual-fluorescent reporter assay. In HeLa cells, the majority of nuclear Rev mutants had activities similar to wild-type Rev. The activities of Rev mutants with abnormal cytoplasmic localization ranged from moderately impaired to nonfunctional. There was no linear correlation between subcellular distribution and levels of Rev activity. In astrocytes, nuclear Rev mutants showed similar impaired activities as the cytoplasmic wild-type Rev. Our data suggest that steady-state subcellular localization is not a primary regulator of Rev activity but may change as a secondary consequence of altered Rev function. The methodologies described here have potential for studying the significance of subcellular localization for functions of other regulatory factors. PMID- 16368435 TI - E-cadherin modulates Wnt-dependent transcription in colorectal cancer cells but does not alter Wnt-independent gene expression in fibroblasts. AB - E-cadherin mediates homophilic adhesion of epithelial cells and is a major determinant of epithelial differentiation during embryonic development and tumor progression. At cell junctions, E-cadherin associates with beta-catenin, which also functions as a transcriptional co-activator of the canonical Wnt signaling pathway by interacting with TCF transcription factors. Here, we have analyzed whether E-cadherin plays a role in the control of gene expression in Wnt dependent and -independent cellular systems. In DLD-1 colorectal cancer cells, which show constitutive activation of Wnt signaling and exhibit E-cadherin-based cell contacts, the siRNA-mediated knock-down of E-cadherin led to the disturbance of cell junctions, translocation of beta-catenin to the nucleus and an enhancement of beta-catenin/TCF-dependent reporter activity. In L929 fibroblasts, which are deficient in Wnt signaling and E-cadherin-mediated cell adhesion, ectopic expression of E-cadherin induced the stabilization of beta-catenin at the cell junctions and caused marked alterations in cellular morphology and phenotype. However, E-cadherin did not significantly change the transcriptional program of these cells as revealed by DNA microarray analysis. Our data indicate that E-cadherin may modulate Wnt-dependent gene expression by regulating the availability of beta-catenin but has a surprisingly small impact on gene expression in the absence of Wnt signaling. PMID- 16368436 TI - Involvement of the ubiquitin pathway in decreasing Ku70 levels in response to drug-induced apoptosis. AB - Ku70 plays an important role in DNA damage repair and prevention of cell death. Previously, we reported that apoptosis caused a decrease in cellular Ku70 levels. In this study, we analyzed the mechanism of how Ku70 levels decrease during drug induced apoptosis. In HeLa cells, staurosporin (STS) caused a decrease in Ku70 levels without significantly affecting Ku70 mRNA levels. We found that Ku70 protein was highly ubiquitinated in various cell types, such as HeLa, HEK293T, Dami (a megakaryocytic cell line), endothelial, and rat kidney cells. An increase in ubiquitinated Ku70 protein was observed in apoptotic cells, and proteasome inhibitors attenuated the decrease in Ku70 levels in apoptotic cells. These results suggest that the ubiquitin-proteasome proteolytic pathway plays a role in decreasing Ku70 levels in apoptotic cells. Ku70 forms a heterodimer with Ku80, which is required for the DNA repair activity of Ku proteins. We also found that Ku80 levels decreased in apoptotic cells and that Ku80 is a target of ubiquitin. Ubiquitinated Ku70 was not found in the Ku70-Ku80 heterodimer, suggesting that modification by ubiquitin inhibits Ku heterodimer formation. We propose that the ubiquitin-dependent modification of Ku70 plays an important role in the control of cellular levels of Ku70. PMID- 16368437 TI - Mechanism of the stress-induced collapse of the Ran distribution. AB - The small GTPase Ran plays a central role in several key nuclear functions, including nucleocytoplasmic transport, cell cycle progression, and assembly of the nuclear envelope. In a previous study, we showed that cellular stress induces the nuclear accumulation of importin alpha, and that this appears to be triggered by a collapse in the Ran gradient, leading to the down-regulation of classical nuclear transport. We report here that a decrease in stress-induced ATP is associated with an increase in cytoplasmic Ran levels. A luciferin-luciferase assay showed that cellular stress decreased the intracellular levels of ATP. Treatment of the cells with ATP-depleting agents altered the distribution of Ran. Furthermore, when exogenous ATP was introduced in oxidative stress-treated cells, a normal distribution of Ran was restored. In addition, a pull-down experiment with an importin beta1 variant that binds to RanGTP showed that oxidative stress was accompanied by a decrease in intracellular RanGTP levels. These findings indicate that the decrease in ATP levels induced by cellular stress causes a decrease in RanGTP levels and a collapse of Ran distribution. PMID- 16368438 TI - Occult ovarian cancer at the time of risk-reducing salpingo-oophorectomy. PMID- 16368439 TI - Intraperitoneal chemotherapy for women with advanced epithelial ovarian carcinoma. PMID- 16368440 TI - Intraperitoneal catheter outcomes in a phase III trial of intravenous versus intraperitoneal chemotherapy in optimal stage III ovarian and primary peritoneal cancer: a Gynecologic Oncology Group Study. AB - OBJECTIVES: To evaluate reasons for discontinuing intraperitoneal (IP) chemotherapy, and to compare characteristics of patients who did versus did not successfully complete six cycles of IP chemotherapy. METHODS: In a phase III trial, women with optimal stage III ovarian or peritoneal carcinoma were randomly allocated to receive IP therapy (paclitaxel 135 mg/m(2) intravenously (IV) over 24 h, cisplatin 100 mg/m(2) IP day 2, paclitaxel 60 mg/m(2) IP day 8) every 21 days for six cycles. Patients unable to receive IP therapy were treated with the alternate (IV) regimen. Variables compared included surgical procedures prior to enrollment, timing of IP catheter insertion, and primary and contributing reasons for discontinuing IP therapy. RESULTS: Among 205 eligible patients randomly allocated to the IP arm, 119 (58%) did not complete six cycles of IP therapy. Forty (34%) patients discontinued IP therapy primarily due to catheter complications and 34 (29%) discontinued for unrelated reasons. Hysterectomy, appendectomy, small bowel resection, and ileocecal resection were not associated with failure to complete six cycles. IP therapy was not initiated in 16% of patients who did versus 5% of those who did not have a left colon or rectosigmoid colon resection (P = 0.015). There was no association between timing of catheter insertion and failure to complete IP therapy. CONCLUSIONS: In this multi institutional setting, it was difficult to deliver six cycles of IP therapy without complications. There appears to be an association between rectosigmoid colon resection and the inability to initiate IP therapy. Catheter choice, timing of insertion, and how surgical treatment of ovarian cancer influences the successful completion of intraperitoneal chemotherapy require further study. PMID- 16368441 TI - Are all aromatase inhibitors the same? A review of controlled clinical trials in breast cancer. AB - BACKGROUND: Five years of tamoxifen therapy has been the standard of care for the adjuvant treatment of estrogen receptor-positive early-stage breast cancer for many years and was the first hormonal treatment for postmenopausal women with advanced or metastatic disease. The third-generation aromatase inhibitors (AIs) anastrozole, exemestane, and letrozole offer new treatment options, although their efficacy has not been compared directly in randomized, double-blind, controlled trials in any breast cancer treatment setting. OBJECTIVE: : The goal of this article was to review the results of recent randomized, controlled clinical trials of the AIs in the settings of neoadjuvant, adjuvant, and advance d/metastatic breast cancer. METHODS: MEDLINE was searched for descriptions of randomized, controlled clinical trials published from 1990 to 2005 using the terms breast cancer, aromatase, aromatase inhibitor, anastrozole, exemestane, and letrozole. Abstracts from the proceedings of several oncology meetings held between 2001 and 2005 were searched to capture relevant emerging data. RESULTS: In 2 Phase III trials comparing an AI with tamoxifen for the adjuvant treatment of breast cancer in postmenopausal women, disease-free survival was significantly improved with anastrozole and letrozole compared with tamoxifen as initial adjuvant treatment (P = 0.01 and P = 0.003, respectively). A switch to either anastrozole (2 Phase III trials) or exemestane (1 Phase III trial) after 2 to 3 years of adjuvant tamoxifen therapy was more effective in reducing the risk of recurrence than continued tamoxifen therapy (P = 0.006, P < 0.002, and P < 0.001, respectively); data on switching to letrozole are expected soon. In another Phase III trial, letrozole was found to improve disease-free survival in the extended adjuvant setting (P < or = 0.001) and was the only AI consistently more effective than tamoxifen in the neoadjuvant setting. In 3 Phase III studies (1 letrozole vs tamoxifen, 2 anastrozole vs tamoxifen), both anastrozole and letrozole were more efficacious than tamoxifen in the first-line setting, and some patients receiving letrozole had better overall response rates compared with those receiving anastrozole in the second-line setting (19.1% vs 12.3%, respectively; P = 0.013). In a patient-preference study, those receiving letrozole reported fewer adverse events than those receiving anastrozole (43% vs 65%; P < 0.003), and more patients preferred letrozole to anastrozole (68% vs 32%; P < 0.01). CONCLUSIONS: Currently, anastrozole and letrozole are associated with the most complete data over the breast cancer care continuum, with efficacy in early-stage, locally advanced, and metastatic disease. In-direct comparisons suggest stronger evidence for the use of letrozole compared with other AIs for breast cancer in postmenopausal women who require estrogen-deprivation therapy. Data from randomized, double-blind comparative studies will help clarify the differences between AIs. PMID- 16368442 TI - Bupropion for major depressive disorder: Pharmacokinetic and formulation considerations. AB - BACKGROUND: Major depressive disorder (MDD) is a common psychiatric condition, with 6.6% of the adult population in the United States experiencing a major depressive episode during any given year. Depressed patients must receive adequate treatment to maximize the likelihood of clinical success. Bupropion hydrochloride, a noradrenergic/dopaminergic antidepressant, is available in 3 oral formulations: immediate release (IR) (given TID), sustained release (SR) (given BID), and extended release (XL) (given QD). Understanding the pharmacokinetic (PK) properties and formulations of bupropion can help optimize clinical use. OBJECTIVES: : The aims of this article were to provide a review of the PK properties of bupropion and identify its various formulations and clinical applications to help optimize treatment of MDD. METHODS: : In this review, data concerning PK trials/reports were collected from articles identified using a PubMed search. The search was conducted without date limitations and using the search terms bupropion, bupropion SR, bupropion XL, bupropion pharmacokinetics, bupropion metabolism, and bupropion drug interactions. Additional reports were selected from references that appeared in articles identified in the original search. In addition, data from studies summarized in product information and labeling were obtained. All available information, concentrating on studies in humans, pertinent to bupropion PK properties and/or formulations was included. RESULTS: : Bupropion is extensively metabolized by the liver (t(1/2), approximately 21 hours). Hydroxybupropion, the primary active metabolite (t(1/2), approximately 20 hours), is formed by cytochrome P450 (CYP) 2B6. At steady state, C(max) of hydroxybupropion is 4- to 7-fold higher, and the AUC is approximately 10-fold greater, compared with those of the parent drug. Threohydrobupropion and erythrohydrobupropion (mean [SD] t(1/2) values, approximately 37 [13] and approximately 33 [10] hours, respectively), the other active metabolites of bupropion, are formed via nonmicrosomal pathways. Relative to bupropion, the C(max) values are approximately 5-fold greater for threohydrobupropion and similar for erythrohydrobupropion. Based on a mouse antitetrabenazine model, hydroxybupropion is approximately 50% as active as bupropion, and threohydrobupropion and erythrohydrobupropion are approximately 20% as active as bupropion. Bupropion lowers the seizure threshold and, therefore, concurrent administration with other agents that lower the seizure threshold should be undertaken cautiously. Potential interactions with other agents that are metabolized by CYP2B6 should be considered. In addition, bupropion inhibits CYP2D6 and may reduce clearance of agents metabolized by this enzyme. Absorption of the XL formulation is prolonged compared with the IR and SR formulations (T(max), approximately 5 hours vs approximately 1.5 and approximately 3 hours, respectively). Bupropion is dosed without regard to food. CONCLUSIONS: : Understanding the PK profile and formulations of bupropion can help optimize clinical use. Bupropion is metabolized extensively, resulting in 3 active metabolites. This metabolic profile, various patient factors (eg, age, medical illnesses), and potential drug interactions should be considered when prescribing bupropion. The 3 formulations-bupropion, bupropion SR, and bupropion XL-are bioequivalent and offer options to optimize treatment for patients with MDD. PMID- 16368443 TI - The spectrum of irritable bowel syndrome: A clinical review. AB - BACKGROUND: Irritable bowel syndrome (IBS) is associated with a substantial burden on individual patients, health care systems, and society as a whole. OBJECTIVES: This review article provides an overview of the disease state and discusses treatment options. METHODS: The MEDLINE database was searched for original research and review articles published in English (from 1966 to July 2005) using the medical subject heading irritable bowel syndrome and the following subheadings: diagnosis, economics, pathophysiology, and therapy. Preference was given to articles that focused on new insights regarding pathophysiology and to those that addressed global symptom improvement for IBS. RESULTS: Symptoms of IBS vary with each patient and disease episode. Although improvement in individual IBS symptoms is desirable, the overall goal in managing patients with IBS is to achieve global symptom improvement. Traditional IBS therapies (ie, psychotherapy/behavioral therapy, bulking agents, antidiarrheals, antispasmodics, and tricyclic antidepressants) have lacked demonstrable efficacy in randomized controlled trials. Recent improvements in the understanding of IBS pathophysiology, particularly regarding the role of neurotransmitters and hormones in gastrointestinal motility, secretion, and visceral perception, have resulted in the development of novel treatments for IBS. In clinical trials, IBS specific treatments (ie, agents that affect serotonergic pathways) have been associated with significant improvements in global symptoms. However, the added cost of treatment with these agents and the potential risks of serious adverse events (eg, severe constipation, severe diarrhea, ischemic colitis) may preclude their widespread use. Initial cost-effectiveness analyses with the novel IBS specific agents suggest the benefits may outweigh the added costs. From a managed care perspective, the costs incurred by IBS-specific therapies may be offset by subsequent reductions in health care resource utilization by patients with IBS. CONCLUSIONS: IBS-specific agents offer an opportunity for patients with IBS to achieve global symptom improvement. However, when weighing the costs and potential risks against the potential benefits, clinicians should consider the nature and severity of the patient's symptoms, the degree of functional impairment, and the presence of psychosocial comorbidities. PMID- 16368444 TI - A review of intermittent subcutaneous apomorphine injections for the rescue management of motor fluctuations associated with advanced Parkinson's disease. AB - BACKGROUND: As Parkinson's disease (PD) progresses,despite optimized pharmacotherapy, patients experience more frequent fluctuations between symptomatic improvement ("on" times) and the return of motor features ("off" times). Apomorphine, the first injectable dopamine agonist available in the United States, is indicated for the acute treatment of "off" episodes (eg, end-of dose wearing-off episodes, unpredictable "on/off" episodes) in patients with advanced PD who are receiving medically optimal antiparkinsonian therapy. OBJECTIVE: This article reviews the pharmacology,clinical efficacy, and tolerability of intermittent subcutaneous apomorphine injections for the management of "off" episodes in patients with PD. METHODS: MEDLINE (1966-July 2005), the Cochrane Database of Systematic Reviews, and International Pharmaceutical Abstracts (1970-July 2005) were searched for original research and review articles published in English. The search terms were apomorphine and Parkinson's disease. The reference lists of articles were also consulted, as was selected information provided by the manufacturer of apomorphine. All relevant identified studies on intermittent subcutaneous administration of apomorphine were included in the review; trials of continuous subcutaneous infusion and non subcutaneous administration of apomorphine were excluded. RESULTS: Intermittent subcutaneous administration of apomorphine produced consistent rescue from "of" episodes in patients with advanced PD, with a symptomatic motor improvement between the predose "off" state and postdose "on" state similar to that achieved with levodopa. The onset of effect occurred within 20 minutes, and the duration of effect was approximately 100 minutes. The therapeutic rescue dose ranged from 2 to 6 mg. During the clinical development program for subcutaneously injected apomorphine, patients required a mean of approximately 3 rescue doses per day. Common adverse effects occurring in > or =20% of patients were injection-site reaction, yawning, dyskinesias, drowsiness, nausea and vomiting, dizziness or postural dizziness, and rhinorrhea. CONCLUSIONS: The available clinical studies indicate that apomorphine is effective in providing prompt and consistent rescue from "off" episodes in patients with PD. Antiemetic prophylaxis and close medical supervision are recommended when initiating apomorphine therapy. PMID- 16368445 TI - Cinacalcet: An oral calcimimetic agent for the management of hyperparathyroidism. AB - BACKGROUND: Uncontrolled hyperparathyroidism (HPT), particularly HPT resulting from chronic kidney disease (CKD), is associated with significant morbidity and cardiovascular mortality. Traditional medical therapy (eg, vitamin D sterols, calcium, phosphate binders) has been inadequate for the management of HPT and its vascular and skeletal complications. OBJECTIVE: : The goal of this article was to review the efficacy and safety profile of cinacalcet, a second-generation calcimimetic, in the management of HPT secondary to CKD, primary HPT, and parathyroid carcinoma. METHODS: MEDLINE, Web of Science, and International Pharmaceutical Abstracts were searched from 1995 to July 2005 using the terms cinacalcet, AMG 073, KRN 1493, calcimimetics, hypercalcemia, and hyperparathyroidism. RESULTS: Compared with placebo, cinacalcet significantly reduced parathyroid hormone levels within 2 to 4 hours after administration (P < 0.05). In Phase III trials involving 1136 patients with secondary HPT, 56% of those who received cinacalcet achieved the National Kidney Foundation Kidney Disease Outcomes Quality Initiative target of a reduction in parathyroid hormone to <300 pg/mL, 65% achieved a calcium-phosphorus product <55 mg2/dL2, and a respective 49% and 46% achieved normalized serum calcium and phosphorus levels (P < 0.001). Cinacalcet's effects were similar regardless of patients' demographic characteristics, duration or mode of dialysis, severity of HPT, or use of concomitant medical therapy. Preliminary evidence suggests that cinacalcet may reverse cortical bone loss. Cinacalcet was well tolerated, with nausea (31%) and vomiting (27%) being the most commonly reported adverse effects. Hypocalcemia was transient in 5% of patients, was usually asymptomatic, and was corrected by dose reduction. CONCLUSIONS: Based on the available evidence, cinacalcet is effective and well tolerated in the treatment of secondary HPT and refractory parathyroid carcinoma. Its use in primary HPT appears promising. Further investigations are needed to determine if cinacalcet can prevent the long-term complications of HPT and reduce mortality. PMID- 16368446 TI - Comparison of the efficacy of ciclesonide 160 microg QD and budesonide 200 microg BID in adults with persistent asthma: a phase III, randomized, double-dummy, open label study. AB - OBJECTIVE: The efficacy of ciclesonide 160 mug QD (given either in the morning or evening) was compared with budesonide 200 mug BID in adults with stable asthma that was pretreated with inhaled corticosteroids. METHODS: This was a randomized, 3-arm, parallel-group study comparing ciclesonide (given in a double-blind, double-dummy regimen) with open-label budesonide. After 2 to 2.5 weeks, during which patients were treated with budesonide 200 microg BID, patients (n = 405) were randomly assigned to receive ciclesonide 160 microg QD AM or 160 microg QD pm, or budesonide 200 microg BID (all administered by metered-dose inhaler) for 12 weeks. All patients received 2 puffs of medication (or placebo) in the morning and evening. The primary efficacy variable was the difference in spirometric forced expiratory volume in 1 second (FEV(1) in liters) from randomization to study end. Secondary efficacy end points were forced vital capacity, peak expiratory flow by spirometry, and diary assessments of peak expiratory flow, asthma symptoms, and rescue medication use. Adverse events were assessed by patient report, investigator observation, physical examination, and laboratory testing; events were classified as mild, moderate, or severe. RESULTS: Baseline demographic characteristics with regard to sex, age, weight, smoking status, baseline medication use, and FEV(1) were balanced among the treatment groups. Over the course of treatment, both ciclesonide and budesonide maintained FEV(1) compared with baseline. Both ciclesonide regimens were as effective as budesonide 200 microg BID in maintaining FEV(1) during the treatment period versus baseline (ciclesonide 160 microg QD am: 95% CI, -0.120 to 0.045 vs budesonide; P = NS; ciclesonide 160 microg QD pm: 95% CI, -0.061 to 0.105 vs budesonide; P = NS). Ciclesonide 160 microg QD (morning or evening) was comparable with budesonide 200 microg BID for maintaining pulmonary function, asthma symptom scores, and rescue medication use. The incidence of adverse events was not significantly different among the treatment groups, and most adverse events were not related to study medication. CONCLUSIONS: In this study, ciclesonide 160 microg QDwas as effective as budesonide 200 microg BID (400 microg total daily dose) in these adults with persistent asthma. Both treatments were well tolerated. PMID- 16368447 TI - Effects of antioxidant supplementation on postprandial oxidative stress and endothelial dysfunction: a single-blind, 15-day clinical trial in patients with untreated type 2 diabetes, subjects with impaired glucose tolerance, and healthy controls. AB - BACKGROUND: Increased generation of reactive oxygen species (ROS) and oxidative stress may be of crucial importance in the pathogenesis of endothelial damage. Furthermore, there is understood to be a relationship between endothelial damage, glycemic control, disorders of lipid metabolism, and coagulative hemostatic disorders. OBJECTIVE: This study investigated within- and between-group changes in various circulating markers of oxidation-reduction balance and endothelial function after a balanced moderate-fat meal with and without antioxidant supplementation in patients with early-stage, untreated type 2 diabetes mellitus; subjects with impaired glucose tolerance (IGT); and healthy controls. METHODS: In this single-blind, controlled clinical study, groups of patients with type 2 diabetes and subjects with IGT were identified and compared with a group of healthy controls. All groups followed a controlled, well-balanced diet for 10 days before and throughout the study. Before and after consumption of a standardized moderate-fat meal, plasma levels of oxidants (malondialdehyde, 4 hydroxynonenal, oxidized low-density lipoprotein), the antioxidant glutathione peroxidase, and markers of endothelial function (NO, endothelin-1, von Willebrand factor [vWF], vascular cell adhesion molecule-1 [VCAM-1]) were determined. These measures were then reassessed after 15 days of standard antioxidant treatment consisting of a thiol-containing antioxidant (N-acetylcysteine 600 g/d), a bound antioxidant (vitamin E 300 g/d), and an aqueous phase antioxidant (vitamin C 250 mg/d). The efficacy of antioxidant treatment in reversing abnormalities in oxidation-reduction balance after a moderate-fat meal was assessed by evaluating changes in plasma levels of ROS on the morning of the 16th day following an overnight fast. Safety was monitored in terms of adverse events, vital signs, physical findings, and laboratory values. RESULTS: The study included 46 patients with type 2 diabetes (23 men, 23 women; mean [SD] age, 41 [3] years; mean body mass index [BMI], 24 [2] kg/m(2)), 46 with IGT (23 men, 23 women; mean age, 39 [3] years; mean BMI, 23 [3] kg/m(2)), and 46 control subjects (23 men, 23 women; mean age, 40 [1] years; mean BMI, 22 [1] kg/m(2)). Before supplementation, all 3 groups had significantly increased levels of oxidants, vWF, and VCAM-1 (all, P < 0.001) and significantly decreased levels of antioxidants and NO (both, P < 0.001) after consumption of a moderate-fat meal. After 15 days of antioxidant treatment, significant improvements in these measures were seen in all groups (P < 0.05). CONCLUSIONS: This study showed changes in oxidation-reduction balance, NO bioavailability, and nonthrombogenic endothelial factors after a moderate-fat meal in patients with type 2 diabetes and those with IGT, but these postprandial changes were reverse in all subjects after 15 days of standard antioxidant supplementation. These findings suggest that the use of anti-oxidants may have decreased oxidative stress in these subjects. PMID- 16368448 TI - Association between human atrial natriuretic peptide Val7Met polymorphism and baseline blood pressure, plasma trough irbesartan concentrations, and the antihypertensive efficacy of irbesartan in rural Chinese patients with essential hypertension. AB - BACKGROUND: Individual variations in the pharmacokinetics and pharmacodynamics of antihypertensive drugs are influenced by genetic and environmental factors. The ANP gene, which encodes the precursor of atrial natriuretic peptide (ANP), is among the candidate genes for genetic susceptibility to hypertension. OBJECTIVE: This study examined the relationship between ANP Val7Met polymorphism (Single Nucleotide Polymorphism database ID: rs5063) and baseline blood pressure (BP), plasma trough irbesartan concentrations, and the antihypertensive efficacy of irbesartan in rural Chinese patients with essential hypertension. METHODS: Patients with essential hypertension who had taken no antihypertensive medications within 4 weeks of study initiation received oral irbesartan 150 mg/d for 4 weeks. Genotyping was performed for all patients. BP was measured before dosing on the 1st and 28th days of treatment. Plasma irbesartan concentrations were measured using high-performance liquid chromatography with fluorescent detection. Antihypertensive efficacy was defined as attainment of a diastolic BP (DBP) <90 mm Hg (DBP analysis), a systolic BP (SBP) <140 mm Hg (SBP analysis), and a DBP <90 mm Hg and SBP <140 mm Hg (DBP and SBP analysis). RESULTS: The study included 756 patients, 621 with the Val/Val genotype and 135 with the Val/Met+Met/Met genotypes. There were no significant differences in age, body mass index, sex, education level, occupation, alcohol consumption, or smoking status between the 2 groups. Patients with the Val/Met+Met/Met genotypes had a significantly lower mean baseline DBP compared with those with the Val/Val genotype (adjusted regression coefficient [SE]: -2.5 [1.0] mm Hg; P = 0.012) and significantly lower mean steady-state plasma trough irbesartan concentrations (adjusted regression coefficient: -12.6 [4.1]; P = 0.002). No significant association was found between antihypertensive efficacy and Val7Met polymorphism in the overall population, but in an analysis by baseline DBP status, patients with the Val/Met+Met/Met genotype a baseline DBP > or =100 mm Hg had significantly smaller reductions in DBP (adjusted regression coefficient: -5.7 [1.4] mm Hg; P < 0.001) and SBP compared with those with the Val/Val genotype and a baseline DBP > or =100 mm Hg (adjusted regression coefficient: -9.8 [2.9] mm Hg; P < 0.001). CONCLUSION: The findings of this study suggest that in these rural Chinese patients with essential hypertension, ANP Val7Met polymorphism may be a genetic marker for baseline DBP, plasma irbesartan concentrations, and the antihypertensive efficacy of short-term irbesartan therapy. PMID- 16368449 TI - Early intervention in migraine with sumatriptan tablets 50 mg versus 100 mg: a pooled analysis of data from six clinical trials. AB - BACKGROUND: In clinical trials evaluating sumatriptan in the treatment of moderate or severe migraine pain, the 50- and 100-mg doses have been comparably effective and well tolerated. OBJECTIVE: To assess the dose-efficacy relationship of sumatriptan tablets given early for mild pain, data from 6 randomized, double blind, placebo-controlled, early-intervention studies of sumatriptan tablets 50 mg and 100 mg (5 of which have been published) were pooled for analysis. These constitute all the studies conducted to date of sumatriptan tablets prospectively given early for mild pain. METHODS: The primary efficacy end point in all the studies was the proportion of patients reporting a pain-free result (ie, mild, moderate, or severe pain reduced to none) 2 hours postdose. Other efficacy end points included the proportion of patients who were migraine free (ie, no pain and no associated symptoms of nausea, vomiting, photophobia, or phonophobia) 2 hours postdose; the proportion reporting worsening of pain (ie, moderate or severe pain) 2 hours postdose; and the proportion with a sustained pain-free result (ie, pain free from 2-24 hours postdose with no use of a second dose of study medication or of rescue medication). Tolerability was assessed by evaluating the incidence of individual adverse events. The investigators assessed each adverse event's relationship to study medication. RESULTS: The number of patients in the intent-to-treat population was 2297 (771 sumatriptan 50 mg, 759 sumatriptan 100 mg, 767 placebo). Patients' mean age ranged from 39.4 to 39.8 years across groups, and most patients were female (90%-92%) and white (89%-90%). A pain-free result 2 hours post dose was reported by significantly more patients who took either dose of sumatriptan tablets compared with placebo and by significantly more patients who took the 100-mg dose compared with the 50-mg dose (50 mg, 49%; 100 mg, 58%; placebo, 24%; P < 0.001, both sumatriptan doses vs placebo, and 100 mg vs 50 mg). A similar pattern was observed for migraine-free results 2 hours postdose (50 mg, 42%; 100 mg, 47%; placebo, 20%; P < 0.05, both sumatriptan doses vs placebo, and 100 mg vs 50 mg), worsening of pain 2 hours postdose (50 mg, 26%; 100 mg, 21%; placebo, 46%; P < 0.05, both sumatriptan doses vs placebo, and 100 mg vs 50 mg), and sustained pain-free results from 2 through 24 hours postdose (50 mg, 30%; 100 mg, 35%; placebo, 12%; P < 0.05, both sumatriptan doses vs placebo, and 100 mg vs 50 mg). Both doses of sumatriptan were well tolerated, and no dose-related trends in the incidence of individual drug-related adverse events were observed. CONCLUSIONS: In this analysis of pooled data from 6 clinical trials, sumatriptan tablets 50 mg and 100 mg administered early in a migraine attack while the pain was mild were well tolerated and significantly more effective than placebo. The 100-mg dose of sumatriptan was significantly more effective than the 50-mg dose. PMID- 16368450 TI - Comparison of fixed-dose combinations of telmisartan/hydrochlorothiazide 40/12.5 mg and 80/12.5 mg and a fixed-dose combination of losartan/hydrochlorothiazide 50/12.5 mg in mild to moderate essential hypertension: pooled analysis of two multicenter, prospective, randomized, open-label, blinded-end point (PROBE) trials. AB - BACKGROUND: High incidences of cardiovascular events coincide with a surge in blood pressure (BP) that occurs in the early morning hours at the time of arousal. Thus, control of BP at this time of day, using oral fixed-dose combinations (FDCs) as required, is important in reducing cardiovascular risk in hypertensive patients. OBJECTIVE: The aim of this analysis was to compare the antihypertensive efficacy in the early morning hours and tolerability of oral FDCs of telmisartan/hydrochlorothiazide (HCTZ) (40/12.5 mg [T40/H12.5] and 80/12.5 mg [T80/H12.5]) versus a low-dose FDC of losartan 50 mg/HCTZ 12.5 mg (L50/H12.5). METHODS: Data from 2 similarly designed prospective, randomized, open-label, blinded-end point (PROBE) studies were pooled and analyzed. The studies were conducted at 72 centers across the United States, and 70 centers in Canada, Europe (9 countries), and the Philippines. Adult male and female patients with mild to moderate essential hypertension (24-hour mean ambulatory diastolic BP [DBP], > or =85 mm Hg; seated cuff DBP, 90-109 mm Hg) were enrolled. Patients were randomly assigned to receive T40/H12.5, L50/H12.5, or T80/H12.5, QD (morning) for 6 weeks. Antihypertensive efficacy was assessed using 24-hour ambulatory BP monitoring (ABPM) and cuff sphygmomanometry at trough, performed at baseline and on completion of active treatment. The primary end point was the reduction from baseline in mean ambulatory DBP over the last 6 hours of the dosing interval. Secondary end points included other ABPM- and clinic-derived changes in DBP and systolic BP (SBP), and control and response rates (SBP response defined as 24-hour mean SBP <130 mm Hg and/or reduction from baseline > or =10 mm Hg; DBP response defined as 24-hour mean DBP <85 mm Hg or reduction from baseline > or =10 mm Hg; DBP control defined as 24-hour mean DBP <85 mm Hg). Tolerability was assessed using patient interview, spontaneous reporting, and clinical evaluation. RESULTS: A total of 1402 patients were enrolled(876 men, 525 women; mean [SD] age, 53.1 [9.9] years) (T40/H12.5, n = 517; L50/H12.5, n = 518; and T80/H12.5, n = 367). With T40/H12.5, the mean reduction in last-6-hour mean ambulatory DBP was 1.8 mm Hg greater compared with that achieved with L50/H12.5 ( 11.3 [0.4] vs -9.4 [0.4] mm Hg; P < 0.001), and with T80/H12.5, the mean reduction was 2.6 mm Hg greater compared with that achieved with L50/H12.5 (-12.0 [0.4] vs -9.4 [0.4] mm Hg; P < 0.001). Analysis of secondary end points found that greater BP reduction occurred with T40/H12.5 and T80/H12.5 compared with L50/H12.5. ABPM SBP control and response rates were similar between the 3 groups, but the ABPM DBP control and response rates were significantly higher with T80/H12.5 compared with L50/H12.5 (46.6% vs 34.0% [P < 0.002] and 69.4% vs 55.0% [P < 0.001], respectively). Clinic SBP and DBP control and response rates were higher with T40/H12.5 and T80/H12.5 compared with L50/H12.5 (SBP response, 80.4% and 80.8% vs 68.5% [both, P < 0.001]; DBP response, 66.1% and 67.4% vs 54.4% [both, P < 0.001]; DBP control, 56.5% and 56.4% vs 44.1% [both, P < 0.001] ). The 2 most commonly recorded adverse events (AEs) were headache (T40/H12.5, 2.9%; L50/H12.5, 3.3%; and T80/H12.5, 3.0%) and dizziness (1.2%, 2.1%, and 3.0%, respectively). Most AEs were mild to moderate. CONCLUSIONS: The results of this pooled analysis of2 PROBE studies in adult patients with mild to moderate essential hypertension suggest that T40/H12.5 and T80/H12.5 conferred greater DBP and SBP control compared with low-dose L50/H12.5, including during the last 6 hours of the dosing interval. All 3 treatments were well tolerated. PMID- 16368451 TI - Preventive drug use in patients with a history of nonfatal myocardial infarction during 12-year follow-up in The Netherlands: a retrospective analysis. AB - BACKGROUND: Myocardial infarction (MI) is a common cause of death in developed countries. Long-term preventive pharmacotherapy has been shown to decrease mortality and morbidity after MI. Based on a literature search, studies of these therapies to date have estimated the use of monotherapy, whereas many patients are prescribed combination therapy. Thus, assessment of long-term combination drug use after MI is timely. OBJECTIVE: The aim of this study was to assess the use of oral antithrombotics, beta-blockers, angiotensin-converting enzyme (ACE) inhibitors, hydroxymethyl-glutaryl coenzyme A reductase inhibitors ("statins"), and their combinations after MI at discharge and during 12-year follow-up. METHODS: This community-based, retrospective data analysis was conducted at Utrecht University, Utrecht, The Netherlands. Data from patients aged > or =18 years at hospital admission who experienced nonfatal acute MI between 1991 and 2000 and had a duration of follow-up > or =30 days were included in the analysis. Data were retrieved from the Pharmo Record Linkage System database, which links pharmacies' dispensation records to hospitals' discharge records on an individual patient level, allowing the investigator to observe individual patients' medication use over time. Primary outcome measures were the use of preventive medicines (oral antithrombotics, beta-blockers, ACE inhibitors, and statins) at discharge, overall use, and persistence during 12-year follow-up. RESULTS: Of 330,000 patients in the database, 4007 were included in the analysis (2828 men, 1179 women; mean [SD] age, 63.5 [12.5] years). Use at discharge and overall use of oral antithrombotics and statins increased significantly between 1991 and 2000, whereas use of beta-blockers and ACE inhibitors increased mainly in patients discharged in the latter years of the follow-up period. Therapy with any combination of drugs increased strikingly from 1991 to 2000, from 47% to 90%. At 1 year after discharge, 32% of patients had discontinued their first-prescribed combination treatments. At 5 years after discharge, this rate increased to 57%, suggesting a low rate of persistence CONCLUSIONS: Based on the results of this retrospective data analysis, the use of MI-preventive drug treatment at and after discharge increased significantly in this population in The Netherlands during the 1990s. Combination therapy increased strikingly. However, persistence with combination therapy was low. PMID- 16368452 TI - Controlled-release oxycodone-induced seizures. AB - BACKGROUND: The use of the opioid oxycodone hydrochloride in the management of chronic pain is gaining popularity principally because of its tolerability. However, opioid-related seizure in patients with epilepsy or other conditions that may decrease seizure threshold has been described in the literature; in particular, oxycodone has been associated with seizure in a patient with acute renal failure. OBJECTIVE: The aim of this article was to report a patient with a history of seizures but normal renal and hepatic function who developed seizure on 2 occasions after oxycodone ingestion. METHODS: A 54-year-old male patient presented with a history of tonic-clonic seizures that developed immediately after intracranial surgery. Long-term treatment with carbamazepine 400 mg QD was started, and the patient was free of convulsions for approximately 7 years. The patient presented to us with severe headache that was nonresponsive to an NSAID and the opiate agonist tramadol. Treatment with controlled-release (CR) oxycodone and tramadol drops (50 mg QID if necessary) was started, and tonic-clonic seizures developed 3 days later. RESULTS: Based on laboratory analysis, the patient had normal renal and hepatic function. On discontinuation of oxycodone treatment, the seizures resolved. However, due to effective pain relief with oxycodone, the patient decided to continue treatment, and seizures recurred. Carbamazepine was then administered 4 hours before oxycodone dosing, which allowed continuation of treatment without seizure. CONCLUSIONS: A patient with a history of seizures controlled with long-term carbamazepine therapy developed seizures when he started treatment with oxycodone CR at recommended doses. Oxycodone CR should be used with extreme caution in patients with epilepsy or other conditions that may decrease seizure threshold. PMID- 16368453 TI - Use of Monte Carlo simulation to assess the pharmacodynamics of beta-lactams against Pseudomonas aeruginosa infections in children: a report from the OPTAMA program. AB - BACKGROUND: Assessing the likelihood of achieving bactericidal pharmacodynamic exposures against Pseudomonas aeruginosa with intravenous antimicrobial regimens would provide insights into the selection of empiric therapy in the pediatric population. OBJECTIVE: The objective of this study was to use pharmacodynamic modeling to determine the likelihood of various pediatric antibiotic regimens achieving bactericidal exposures against P aeruginosa in children. METHODS: Minimum inhibitory concentrations (MICs) were determined for meropenem (20 and 40 mg/kg q8h), imipenem (15 and 25 mg/kg q6h), ceftazidime (50 mg/kg q8h), cefepime (50 mg/kg q8h), and piperacillin/tazobactam (75 mg/kg q6h) against P aeruginosa isolates from 2 pediatric institutions. A 5000-patient Monte Carlo simulation was performed to predict attainment of pharmacodynamic targets against P aeruginosa for each of these regimens in a population of 10-year-olds. Optimal regimens were defined as those that had a > or =90% likelihood of attaining target exposures. RESULTS: At institution 1, high-dose imipenem, high-dose meropenem, and ceftazidime achieved bactericidal pharmacodynamic exposures (likelihood of target attainment: 94%, 92%, and 92%, respectively). No other regimen was associated with a high probability of attaining bactericidal exposure (low-dose imipenem, 87%; cefepime, 85%; low-dose meropenem, 84%; piperacillin/tazobactam, 60%). At institution 2, no regimen was associated with a high likelihood of attaining bactericidal exposure; the calculated probabilities were cefepime, 78%; ceftazidime, 65%; high-dose meropenem, 58%; high-dose imipenem, 57%; low-dose imipenem, 54%; low-dose meropenem, 47%; and piperacillin/tazobactam, 47%. A lack of agreement between attainment of bactericidal exposures and percent susceptibility was apparent for many of the regimens. CONCLUSIONS: Few regimens demonstrated a high likelihood of achieving bactericidal exposures against P aeruginosa at these institutions. Importantly, percent susceptibility overestimated attainment of the bactericidal target for some regimens, suggesting that further study is necessary in pediatric patients. The findings of this study highlight differences in target attainment and MIC distributions between institutions, emphasizing the importance of using institution-specific data when selecting empiric antimicrobial therapy. PMID- 16368454 TI - Evidence gaps in complementary and alternative medicine use. PMID- 16368455 TI - Evaluation of the cost-effectiveness of sirolimus versus cyclosporin for immunosuppression after renal transplantation in the United Kingdom. AB - OBJECTIVE: The purpose of this study was to evaluate the cost-effectiveness of sirolimus compared with cyclosporin for the postsurgical management of renal transplant recipients, from the perspective of the UK National Health Service and the Personal Social Service. METHODS: A discrete event stochastic simulation model was developed to evaluate both cost-effectiveness and cost utility over 10 and 20 years after transplant using historical data on 937 renal transplant recipients from the University Hospital of Wales in Cardiff, United Kingdom. The simulation was designed to forecast the incidence of acute rejection events, graft failure, retransplant, frequency of hemodialysis (HD) and peritoneal dialysis (PD), and death. Cox proportional hazard models were derived from historical transplant data, in which 1-, 2-, and 3-year post-transplant serum creatinine levels were used as the key drivers for predicting graft success and survival. Costs were reported as year-2003 UK pounds sterling (1 UK pound = US $1.76). Probabilistic sensitivity analysis was conducted and results reported with particular attention to 2 threshold values, 30,000/QALY and 20,000/QALY RESULTS: Over a 10-year time horizon, treatment with sirolimus was projected to produce a gain of 0.60 discounted year of functioning graft with a cost savings of 276 UK pounds per patient. Over a 20-year time horizon these benefits increased to 1.59 discounted years of functioning graft and a cost savings of 7405 UK pounds per patient. Using sensitivity analysis of the 10-year model, the only factors found to cause the probability of exceeding a 30,000 ceiling to be >5% were the proportion of subjects maintaining continuous graft function and the use of low-dose cyclosporin. With the 20-year model, sirolimus maintained cost effectiveness across most scenarios in sensitivity analysis. CONCLUSIONS: In this model analysis, sirolimus was cost-effective compared with cyclosporin for 10 to 20 years after renal transplantation in the United Kingdom, from the perspective of the UK National Health Service and Personal Social Service. PMID- 16368456 TI - Herb and supplement use in the US adult population. AB - BACKGROUND: Research on the scope of use and factors associated with herbal medicine use is limited. OBJECTIVES: The aims of this work were to assess national usage patterns, reasons for use, and the perceived efficacy of herbal products and dietary supplements. METHODS: This was a secondary analysis of the complementary and alternative medicine (CAM) supplement to the 2002 National Health Interview Survey (NHIS), conducted by the National Center for Health Statistics (NCHS). Participants were asked whether they had used natural herbs for their own health and treatment. Those who responded yes were compared with those who responded no. Supplement users were asked whether they had used any of 36 specific herbs or nonherbal dietary supplements (eg, glucosamine, fish oil, bee pollen), how important the use of CAM treatment was to them, whether they had seen a CAM provider, and whether they had informed a conventional medical provider about their use. NCHS weights, derived from Decennial Census data, were used to calculate national prevalence estimates. Group comparisons of herbal use were conducted with the Wald x(2) test. RESULTS: A total of 31,044 adults participated in the 2002 NHIS CAM survey; 632 were omitted from analyses due to incomplete information. In all, 5787 adults said they had used herbs or supplements during the previous 12 months, of whom 57.3% said they used these products to treat specific conditions. Based on these responses, an estimated approximately 38.2 million adults in the United States used herbs and supplements in 2002. More than half of all users said that herbs and natural products were important to their health and well-being. Use rates were higher for women than men (21.0% vs 16.7%; P < 0.001); adults aged 45 to 64 years (P < 0.001 vs other age groups); those of multiple races (32.2%), Asians (24.6%), or American Indians or Alaskan natives (21.9%) rather than whites (19.1%) or blacks (14.3%) (effect of race, P < 0.001); residents of the western United States (effect of region, P < 0.001), and college graduates (25.3% vs 10.4% among those who did not graduate high school; effect of education, P < 0.001). Only 33.4% told a conventional health care provider about their herb or supplement use use. CONCLUSION: Herb and natural supplement use was widespread in the US adult population in 2002, according to data from the NHIS CAM survey, despite the fact that few participants informed their conventional health care providers about such use. PMID- 16368457 TI - Leptospira DNA detection for the diagnosis of human leptospirosis. AB - This study aims to analyse PCR applicability to the diagnosis of human leptospirosis and to compare the sensitivity of two primer pairs in urine and blood samples. PCR with G1/G2 and LP1/LP2 primers was specific and able to detect 10pg of DNA by agarose gel and 1pg by hybridization. Twenty-one serovars, representing 20 serogroups of pathogenic leptospires, were amplified with G1/G2 primers. DNA from two non-pathogenic serovars, Andamana and Patoc, was not amplified. For hybridization, one probe employing DNA from most prevalent leptospires (serovars Icterohaemorrhagiae, Copenhageni, and Autumnalis) was chosen in accordance with the microagglutination titres in patient samples. It was observed that not all serovars hybridized with the PCR products of G1/G2 and LP1/LP2 primer amplification, suggesting heterogeneity in the sequence amplified by these primers. G1/G2-primed amplifications of blood and/or urine samples were shown to be significantly more sensitive (57.6%) than the LP1/LP2 primers (33.3%), P=0.04, when positivity of patients is considered. When each primer pair and only urine samples were considered, PCR positivity was higher for G1/G2 primers than for LP1/LP2 (P=0.007). G1/G2 presented greater sensitivity in urine than in blood, and LP1/LP2 presented greater sensitivity in blood than in urine, although these differences were not statistically significant. The positivity of PCR per patient using both primers in blood and/or urine samples was 63.6%, with 84.4% efficiency. PCR was useful for patients without microagglutination detectable antibodies, for whom it was able to diagnose nine out of 11 patients (81.8%). PMID- 16368458 TI - JC virus DNA in cerebrospinal fluid samples from Brazilian AIDS patients with focal brain lesions without mass effect. AB - OBJECTIVE: To evaluate the presence of JC virus DNA in CSF samples from Brazilian AIDS patients with focal lesions of CNS white matter without mass effect compatible with progressive multifocal leukoencephalopathy (PML). METHODS: CSF samples from AIDS patients with neurological symptoms and a CT scan showing focal lesions of CNS white matter without mass effect suggestive of PML, and from AIDS and non-AIDS patients with non-PML neurological diseases were tested for JC virus DNA by PCR. The primers used to amplify the T antigen region of the JC virus resulted in a 173-bp fragment. The presence of the JC virus was confirmed by digestion of the PCR product using BamH1. RESULTS: The PCR for JCV DNA was negative in 119/120 non-PML CSF samples (specificity =99.2%). Of 56 CSF samples from AIDS patients with focal lesions of CNS white matter without mass effect, JCV DNA was positive in 48.2% (27/56). In 23/29 (79.3%) JCV DNA-negative cases, other causes for the encephalitic lesions were found. No JCV DNA-positive cases showed other diagnoses. CONCLUSIONS: The prevalence of JCV DNA by PCR in CSF samples from Brazilian AIDS patients with focal brain lesions, without mass effect was 48.2%. In these patients, a negative JCV PCR is highly suggestive of other neurological conditions. PMID- 16368459 TI - The current burden of pneumococcal disease in England and Wales. AB - OBJECTIVE: To evaluate the potential impact of various pneumococcal conjugate vaccination strategies, it is critical to ascertain the pre-vaccination epidemiology and to have a detailed evaluation of the current burden of pneumococcal disease. METHOD: A variety of national data sources and GP sentinel surveillance systems were used to estimate the incidence, number of hospital admissions, deaths, and GP consultations due to pneumococcal disease in England and Wales. Clinical outcomes included pneumococcal meningitis, bacteraemia, pneumonia and otitis media. A statistical model was used to attribute GP consultation recorded as pneumonia and acute otitis media to specific aetiological causes when these were not recorded. RESULTS: The burden of pneumococcal disease is considerable, with incidence rates of both invasive and non-invasive disease peaking in children (<5 years) and in the elderly (75+ years). Around 5800 hospitalisations specifically mentioning Streptococcus pneumoniae are estimated to occur annually in England and Wales, almost 40,000 for lobar pneumonia and over 15,000 for otitis media. There may be an additional 70,000 GP consultations for pneumococcal related community acquire pneumonia and over 630,000 for otitis media. A significant proportion of hospitalisations and GP consultations for pneumococcal disease occur among high-risk groups, with over 80% of hospital admissions reporting more than one diagnosis. PMID- 16368460 TI - Is nitric oxide involved in the tolerance of Calomys callosus as a reservoir host towards Trypanosoma cruzi infection? AB - Trypanosoma cruzi, the agent of Chagas disease, is known to cause enhanced nitric oxide (NO) production, which might be involved in host resistance. The inducible nitric-oxide-synthase (iNOS) is assumed to be responsible for the NO increase after several infections. We studied the potential role of NO in Calomys callosus, a natural reservoir of this protozoan parasite. The concentration of NO was determined in spleen and liver of animals infected with two different T. cruzi strains, BOL and BOL-SB. Furthermore, the iNOS mRNA expression was quantified in the same cell types. NO production was detectable in both tissues exhibiting only slight differences compared to non-infected controls. All measured NO values were significantly lower than those reported for a number of different mouse strains, which displayed extremely enhanced NO levels after T. cruzi infection. Surprisingly, iNOS mRNA expression was induced in infected C. callosus but without subsequent increase of NO levels, indicating a post transcriptional regulation mechanism. In summary, our results, indicate that the tolerance of C. callosus to T. cruzi is only accompanied by non-toxic NO intracellular concentrations. PMID- 16368461 TI - Scrub typhus among hospitalised patients with febrile illness in South India: magnitude and clinical predictors. AB - OBJECTIVES: To derive a clinical algorithm for diagnosis of scrub typhus among patients hospitalized with febrile illness and to determine predictors of bad prognosis. METHODS: Patients hospitalized with febrile illness of 5-30 days duration were evaluated for common aetiologies. Sera were tested for antibodies (IgM and IgG) to Orientia tsutsugamushi using ELISA kit. RESULTS: Among 207 patients, 50 had elevated levels of IgM antibodies. The data of these patients were compared with that of 16 controls having febrile illnesses in whom rickettsial infection was ruled out. Transaminase elevation (>twice normal) was present in 90% and was significantly (P=0.004) more common in those with scrub typhus. If a combination of elevated transaminases, thrombocytopenia and leukocytosis is used, the specificity and positive predictive value are about 80%. Case fatality rate was 14%. Univariate analysis showed that hyperbilirubinemia (>1.5mg%) has a RR of 9 (95% CI=1.48-58.5) and elevated creatinine level (>1.4 mg%) had a RR of 43.99 (95% CI=3.65-530.5) for death. Elevated creatinine level was found to be an independent predictor of mortality (P=0.02). CONCLUSION: In developing countries with limited diagnostic facilities, it is prudent to recommend empiric therapy in patients with undifferentiated febrile illness having evidence of multiple system involvement especially if there is transaminase elevation. Elevated creatinine may predict bad outcome. PMID- 16368462 TI - Geographic and socioeconomic variation in meningococcal disease: a rural/urban comparison. AB - OBJECTIVES: To examine the incidence of meningococcal disease in a defined geographic area and relate this to rural/urban residence and material deprivation. METHODS: Retrospective review of statutory notifications, laboratory records and death registrations from 1 January 1996 through 31 December 2001 in an English health area. Incidence of disease was analysed in two populations residing within the area, one rural and one urban. Deprivation was measured using the Townsend material deprivation score. RESULTS: Altogether, 183 cases of meningococcal disease were identified. Average annual incidence was significantly higher (P=0.019) in the urban area (8.8/10(5)) compared to the rural area (6.2/10(5)). In 1996, the relative risk (RR) was greater in the urban area (RR 3.20 95% CI 1.40-7.32) compared to the rural area. During the study period a significant upward trend in disease incidence in the rural area (chi2 for trend P=0.018) was identified whereas a non-significant decrease was observed in the urban area. In the urban area, but not in the rural area, increasing deprivation was strongly associated with increasing disease incidence (chi2 for trend P=0.00010). CONCLUSIONS: Meningococcal disease places a disproportionate disease burden on certain populations. Further research is required to identify measures to address the inequalities identified. PMID- 16368463 TI - Combination of caspofungin and an azole or an amphotericin B formulation in invasive fungal infections. AB - OBJECTIVES: Combination of caspofungin and another anti-fungal agent raise expectation of improved efficacy in severe fungal infections including failures to first line therapy. METHODS: We assessed the efficacy and safety of a combination therapy including caspofungin in 17 immunosuppressed or postoperative patients progressive despite standard anti-fungal therapy. RESULTS: The infections included aspergillosis (6), invasive candidiasis (9), mucormycosis (1) and Scedosporium pneumonia (1). Infections had failed one to four prior lines of treatment. The anti-fungal agent combined to caspofungin was either an amphotericin B formulation or an azole. There were 12 favourable responses (71%) and five failures. The survival rate at 3 months was 47%. Eleven patients died within 2-533 days. The causes of death included the initial fungal infection (4), relapse of the infection after switching to oral monotherapy (2), breakthrough aspergillosis (1), and the underlying condition (4). Clinical and renal tolerance were good. Significant hepatic abnormalities were recorded in eight (50%) of the 16 patients evaluable for biological tolerance. CONCLUSION: Caspofungin combined with an azole or with amphotericin B may be of interest in the treatment of serious fungal infections after failure of conventional therapy. Close monitoring of hepatic function is required. These approach should be evaluated in prospective trials. PMID- 16368465 TI - Toward understanding the dynamics of membrane-raft-based molecular interactions. AB - The cell membrane is a 2-dimensional non-ideal liquid containing dynamic structures on various time-space scales, and the raft domain is one of them. Existing literature supports the concept that raft dynamics may be important for its formation and function: the raft function may be supported by stimulation induced raft association/coalescence and recruitment of various raftophilic molecules to coalesced rafts, and, importantly, they both may happen transiently. Thus, one must always consider the limited association time of a raft or a raftophilic molecule with another raft, even when one interprets the results of static experiments, such as immunofluorescence and pull-down assays. Critical considerations on the chemical fixation mechanism and immunocolocalization data suggest that the temporary nature of raft-based molecular interactions may explain why colocalization results are sensitive to subtle variations in experimental conditions employed in different laboratories. PMID- 16368466 TI - Menopausal hormone therapy (HT) in patients with breast cancer. AB - OBJECTIVES: To assess the effect of menopausal hormone therapy (HT) on reoccurrence, cancer-related mortality, and overall mortality after a diagnosis of breast cancer. METHODS: We performed a quantitative review of all studies reporting experience with menopausal HT for symptomatic use after a diagnosis of breast cancer. Rates of reoccurrence, cancer-related mortality, and overall mortality were calculated in this entire group. A subgroup analysis was performed in studies using a control population to assess the odds ratio of cancer reoccurrence and mortality in hormone users versus non-users. RESULTS: Fifteen studies encompassing 1416 breast cancer survivors using HT were identified. Seven studies included a control group comprised of 1998 patients. Among the 1416 HT users, reoccurrence was noted in 10.0% (95% CI: 8.4-11.6%). Cancer-related mortality occurred at a rate of 2.6% (95% CI: 1.8-3.7%), while overall mortality was 4.5% (95% CI: 3.4-5.8%). Compared to non-users, patients using HT had a decreased chance of reoccurrence and cancer-related mortality with combined odds ratio of 0.5 (95% CI: 0.2-0.7) and 0.3 (95% CI: 0.0-0.6), respectively. CONCLUSIONS: In our review, menopausal HT use in breast cancer survivors was not associated with increased cancer reoccurrence, cancer-related mortality or total mortality. Despite conflicting opinions on this issue, it is important for primary care physicians to feel comfortable medically managing the increasing number of breast cancer survivors. In the subset of women with severe menopausal symptoms, HT options should be reviewed if non-hormonal methods are ineffective. Future trials should focus on better ways to identify breast cancer survivors who may safely benefit from HT versus those who have a substantial risk of reoccurrence with HT use. PMID- 16368467 TI - Cigarette smoking, estrogen levels, and hot flashes in midlife women. AB - OBJECTIVE: The aims of this study were to examine the association of smoking with the occurrence, frequency, and severity of hot flashes and to determine whether the mechanism by which active cigarette smoking increases the risk of hot flashes is by lowering estradiol and estrone levels. METHODS: A case-control study was conducted among women aged 45-54 years to examine risk factors for hot flashes. Cases were women who reported ever experiencing hot flashes (n = 353). Controls were women who reported never experiencing hot flashes (n = 258). Each participant completed a questionnaire and provided a blood sample that was used to measure estradiol and estrone levels. RESULTS: The results showed that both current and ever smokers had higher odds than never smokers of experiencing any and more severe hot flashes. Further, significant positive associations were observed between frequency and duration of smoking and the experiencing of any and more severe hot flashes. Smoking was not associated with estradiol or estrone levels in univariate analyses. In addition, the odds ratios for the associations between the cigarette smoking variables and hot flashes did not change when the hormone variables were added to the model. CONCLUSIONS: These findings indicate that smoking is associated with the occurrence of any and more severe hot flashes, independent of estrogen levels. PMID- 16368468 TI - Factors associated with determinants of sexual functioning in midlife women. AB - OBJECTIVES: The present study was conducted to assess the associations between a woman's passionate love for her partner and sexual satisfaction and demographic factors, health and life-style characteristics, menopausal status, and menopausal symptoms. METHODS: A cross-sectional study was conducted among women (40-60 years) residing in Maryland (n = 846). chi(2)-tests were performed to evaluate the associations between each of the outcome variables (sexual satisfaction and passionate love for the partner) and predictor/independent variables. Logistic regression analysis was performed to determine whether significant associations from chi(2) analyses remained significant after adjustment for confounders. RESULTS: Older age (adjusted odds ratio (OR) 1.04; 95% confidence interval (CI) 1.01, 1.07), higher education (OR 1.47; 95% CI 1.09, 1.99), and alcohol intake (OR 1.42; 95% CI 1.03, 1.95) were associated with low passion for the partner. Older age was also a significant predictor of low sexual satisfaction (OR 1.04; 95% CI 1.01, 1.07). Women experiencing vaginal dryness had higher odds of low passion for partner (OR 1.67; 95% CI 1.21, 1.31) and low sexual satisfaction (OR 1.58; 95% CI 1.14, 2.20) than women not experiencing vaginal dryness. CONCLUSIONS: Older age, higher education, alcohol intake, and vaginal dryness are significantly associated with lower levels of passionate love for the partner, while older age and vaginal dryness are significantly associated with lower levels of sexual satisfaction in midlife women. PMID- 16368469 TI - Comparison of effects of pravastatin and hormone therapy on soluble P-selectin and platelet P-selectin expression in postmenopausal hypercholesterolemic women. AB - OBJECTIVE: Recent trials have suggested an adverse early effect on cardiovascular risk of hormone therapy (HT) in postmenopausal women, an effect which could be due to an increase in arterial thrombosis via platelet activation. We examined the effect of HT on platelet surface expression of P-selectin, a marker of platelet activation, and plasma levels of soluble P-selectin, also believed to be a marker of platelet activation, and compared these effects with pravastatin, a drug proven to reduce cardiovascular events and reported to decrease both platelet and soluble P-selectin. METHODS: Surface expression of platelet P selectin, soluble P-selectin and fasting lipids were measured at baseline and 6 months in a randomized, double-blind study of postmenopausal hypercholesterolemic women comparing low-dose combined HT (1mg estradiol + 0.5 mg norethisterone acetate; n = 26) with pravastatin (n = 24). RESULTS: After adjusting for baseline levels, HT and pravastatin produced similar reductions in soluble P-selectin (p < 0.0001 for both). The percentage of platelets expressing P-selectin was also reduced by pravastatin (p = 0.025), but there was a trend to an increase in platelet P-selectin expression with HT (p = 0.13), and a significant difference between pravastatin and HT in the changes in platelet P-selectin (p < 0.002). No relationship was evident between changes in soluble or platelet P-selectin and changes in lipids with either treatment. CONCLUSIONS: In postmenopausal hypercholesterolemic women, both pravastatin and HT reduced soluble P-selectin levels, but only pravastatin reduced P-selectin expression on the surface of platelets. An implication of these findings is that the reduction in soluble P selectin by HT may occur by a non-platelet related mechanism. PMID- 16368470 TI - Does the Aging Males' Symptoms scale assess major depressive disorder?: A pilot study. AB - OBJECTIVES: The objectives of the study were to find the prevalence of major depressive disorder (MDD) in male climacteric outpatients in Japan, and to determine whether symptoms on the Aging Males' Symptoms (AMS) scale scores differed between patients with and without MDD, with the aim of increasing the specificity of future symptoms scales for partial androgen deficiency of the aging male (PADAM). METHODS: Eighty-three patients aged 40-70 who visited the male-climacteric services as outpatients were assessed using three items: a self administered questionnaire corresponding to diagnosis for MDD, the Beck Depression Inventory, and the AMS scale. RESULTS: Almost half the patients had MDD. The total AMS score, the scores on the AMS psychological, somatic and sexual subscales, and the scores for all except three questionnaire symptoms were higher in patients with MDD. There were strong correlations between the AMS scale and the Beck Depression Inventory. CONCLUSIONS: There is a higher prevalence of MDD in male-climacteric outpatients and scores on most items of the AMS scale were higher for patients with MDD. We suggest that only those symptoms whose scores did not differ between patients with and without MDD are used to assess symptoms of PADAM in the presence of MDD, or that the current AMS is used only after diagnosis of MDD and elimination of these patients. Then the relevance of each item of the AMS to testosterone levels should be simultaneously examined in the future study, which will determine the items highly specific to PADAM symptoms. PMID- 16368471 TI - Pulsed estrogen therapy improves postmenopausal quality of life: a 2-year placebo controlled study. AB - OBJECTIVE: To investigate the effect of pulsed estrogen therapy S21400 (intranasal 17 beta-estradiol) on different quality of life (QoL) dimensions in early postmenopausal women treated with S21400 150 microg per day, S21400 300 microg per day, or placebo in a double blind, randomized, controlled 2-year study. STUDY DESIGN: QoL was assessed based on the validated Women's Health Questionnaire designed for peri- and post-menopausal women. Three hundred and thirty-five healthy, early postmenopausal Danish women, 53 years of age in average, who completed one questionnaire at baseline and one under study treatment were included in the analysis set. All analyses were performed on an intention-to-treat basis. RESULTS: QoL improved significantly in both S21400 groups compared to placebo in the dimensions 'memory/concentration', 'vasomotor symptoms', 'sleep problems' and 'sexual behavior' (difference in mean change scores being respectively +7.9, +28.3, +9.9 and +10.8%, p < 0.001, between the S21400 300 microg and placebo group). There were no significant differences between actively treated groups and placebo in the dimensions 'anxiety/depressed mood' and 'well-being'. CONCLUSION: Pulsed estradiol therapy had a pronounced effect not only on vasomotor symptoms but also a significant and clinically relevant improvement in several other QoL dimensions. PMID- 16368472 TI - Prevention of postmenopausal bone loss with long-cycle hormone replacement therapy. AB - OBJECTIVE: Postmenopausal bone loss and osteoporotic fractures can be prevented by hormone replacement therapy (HRT). However, opposed HRT may increase the risk of breast cancer above that associated with estrogen alone and in non hysterectomized women estrogen substitution alone increases the risk of uterine cancer, which triggered renewed interest in long-cycle HRT regimens (estrogen replacement therapy with progesterone-free intervals up to 6 months). The effects on bone of such long-cycle HRT regimens are unknown. The objective of the present study was to compare the effects on bone and the endometrium of long-cycle HRT and conventional HRT. METHODS: Seventy-three healthy non-hysterectomized postmenopausal women were randomized to either conventional HRT (estradiol (E2) 2 mg/d during 12 days, E2 2 mg/d plus 1 mg/d of norethisterone acetate (NETA) during 10 days, E2 1 mg/d for 6 days) or long-cycle HRT treatment (two cycles with E2 2 mg/d during 28 days, followed by one cycle of conventional HRT and repeated every 3 months). Primary endpoint was the change in bone mineral density (BMD) at the lumbar spine (LS) over 24 months. RESULTS: BMD at LS increased significantly versus baseline in both treatment groups (conventional HRT +3.8 +/- 0.6%, long-cycle HRT +3.3 +/- 0.5%, p < 0.0001 for both) with no significant difference between treatment groups over 24 months. Similar significant BMD increases versus baseline were observed at the femoral neck, while biochemical markers of bone turnover (osteocalcin and deoxypyridinoline) were significantly decreased over 24 months. There were no endometrial or breast related adverse events reported. CONCLUSION: Long-cycle HRT may be a valid alternative to conventional HRT with regard to protection against postmenopausal bone loss. PMID- 16368473 TI - The hysterectomized woman. Is she special? The women's health in the Lund area (WHILA) study. AB - OBJECTIVE: To delineate the health profile of hysterectomized women and to assess whether women who have undergone hysterectomy have a different health profile even before surgery. MATERIAL AND METHODS: The WHILA project covers all women (n = 10,766) aged 50-60, living in the Lund area and are based on questionnaires and personal interviews tied to laboratory examinations. RESULTS: 6917 women (64.2%) had complete questionnaires and laboratory tests, 800 were hysterectomized (11.6%). Logistic regression analysis revealed that hysterectomized women had more "dizziness" 1.40 (1.19-1.66), "nervous problems" 1.29 (1.07-1.56), "backache" 1.37 (1.16-1.62), "joint problems" 1.29 (1.09-1.52), "eye problem" 1.20 (1.02-1.42) and "headache" 1.17 (1.00-1.37). For both somatic (5.22 versus 4.49 mean value, p < 0.001) and psychological (4.19 versus 3.86 mean value, p = 0.002) symptoms, the number was higher in hysterectomized women. Logistic regression analysis revealed that among hysterectomized women university education was less common odds ratios 0.73 (95% confidence interval 0.58-0.91) as well as working full time 0.76 (0.62-0.93). A higher body weight at the age 25, 1.01 (1.001-1.02) as well as a weight gain of more than 5 kg during the last 5 years 1.27 (1.07-1.50), elevation of serum triglycerides 1.29 (1.16-1.44), high density (HDL) 1.44 (1.14-1.80) and low-density lipoprotein (LDL) cholesterol 1.11 (1.02-1.21) as well as the bone density 1.08 (1.00-1.17). Hysterectomized women had a lower age at giving first birth (p < 0.001), shorter interval between menstrual periods (p < or = 0.001) and less frequent amenorrhic episodes (p < 0.05). The hysterectomized women used IUD to a lesser extent (p < 0.05) but used hormone therapy (HT) (p < 0.001) and utilized health care services (p < 0.001) more often. CONCLUSIONS: Long after surgery, several somatic and psychological symptoms were still more common in hysterectomized women. A low frequency of amenorrhic episodes and lower age at giving first birth, concomitant with a higher body weight already at age 25 may imply that women who end up hysterectomized have a specific health profile long before as well as long after surgery. PMID- 16368474 TI - Territorial waters of the Baltic Sea as a source of infections caused by Vibrio cholerae non-O1, non-O139: report of 3 hospitalized cases. AB - A fatal infection with temporal relation to 2 other febrile infections caused by Vibrio cholerae non-O1, non-O139 (NCV) occurred in Finland in 2003. All infections were associated with contact with seawater. The patient who died had also eaten home-salted whitefish, tested positive for NCV, preceding his symptoms. All patients had compromising factors, and all strains were distinguishable by pulsed-field gel electrophoresis and negative for the ctx gene. These 3 cases illustrate that, despite being uncommon in Finland, NCVs can cause clinically significant and even fatal infections. PMID- 16368475 TI - Pharmacodynamic assessment of the activity of high-dose (750 mg) levofloxacin, ciprofloxacin, and gatifloxacin against clinical strains of Pseudomonas aeruginosa. AB - The objective of this study was to comparatively evaluate specific bacterial killing ability of high-dose (750 mg) levofloxacin, ciprofloxacin, and gatifloxacin against 2 clinical isolates of Pseudomonas aeruginosa (PA-21 and PA 2105). An in vitro pharmacodynamic modeling apparatus was used to expose the P. aeruginosa isolates to total peak concentrations and elimination characteristics associated with each quinolone. All experiments were conducted over 24 h, and a subsequent dose of ciprofloxacin was given at 12 h to emulate twice-daily dosing. Respective 3-log reductions in PA-24 occurred after 0.6, 1.0, and 2.6 h for levofloxacin, ciprofloxacin, and gatifloxacin; regrowth was seen with all 3 agents, but was greatest with gatifloxacin. PA-2105 had 2- to 4-fold higher minimal inhibitory concentrations (MICs) than PA-24. Gatifloxacin failed to achieve a 3-log reduction. Levofloxacin and ciprofloxacin took roughly 3.5 h to decrease initial inoculum by 3 logs, but regrowth of PA-2105 followed. Simulated doses of levofloxacin and ciprofloxacin showed comparable activity against each study isolate; less activity was observed with gatifloxacin. Levofloxacin versus PA-24 was the only regimen that approached the desired AUC/MIC(0-24) ratio of greater than 100-125 and achieved the targeted peak/MIC ratio of > or =8. Although quinolones are typically used in combination with other antibiotics for P. aeruginosa, differences in activity favor the use of levofloxacin or ciprofloxacin for the study isolates. Use of gatifloxacin may contribute to the increased rate of quinolone-resistant P. aeruginosa. PMID- 16368476 TI - Contemporary antimicrobial activity of triple antibiotic ointment: a multiphased study of recent clinical isolates in the United States and Australia. AB - Triple antibiotic ointment (TAO) containing neomycin, polymyxin B, and bacitracin was launched in the 1950s in the United States (USA) as a prescription product and then was used over the counter (OTC) since the 1970s (USA) to prevent superficial wound infections. In Australia, TAO has been restricted to prescription use. This study 1) determined cross-resistance patterns of neomycin compared with other aminoglycosides; 2) determined the level and trend of resistance to TAO and individual components especially versus mupirocin-resistant strains (USA); and 3) established the baseline TAO activity level against pathogens from Australia. A total of 200 strains (> or =50% gentamicin-resistant) from the United States were used for the cross-resistance study including Staphylococcus aureus (110), coagulase-negative staphylococci (CoNS; 50), Pseudomonas aeruginosa (10), Escherichia coli (20), and other Enterobacteriaceae (10) tested against TAO, bacitracin, polymyxin B, neomycin, amikacin, gentamicin, streptomycin, tobramycin, and mupirocin. Fifty gentamicin-resistant isolates from each year (1997-2002) were used to determine the activity of TAO over time. Baseline resistance rates of TAO among 300 Australian isolates (AGARS Program, 2002-2003) were also studied. Reference broth microdilution methods were used in all phases of this study. At a 1:100 dilution of the ointment concentration, TAO inhibited all CoNS, Pseudomonas aeruginosa, and Enterobacteriaceae isolates, and resistance to TAO among Staphylococcus aureus at this concentration was only 5% in the cross-resistance study. Patterns of susceptibility in the United States did not significantly vary from 1997 to 2002. Australian pathogens showed that TAO was 98% active against methicillin-resistant Staphylococcus aureus and 100% for Enterobacteriaceae, methicillin-susceptible S. aureus, CoNS, and P. aeruginosa, the rates equivalent to those observed in the United States. Mupirocin-resistant S. aureus (5%) and CoNS (47%) were all TAO-susceptible. All Gram-negative species were also mupirocin-resistant, but inhibited by neomycin and/or polymyxin B components of TAO. In conclusion, aminoglycoside resistance patterns differ significantly, and none of the commonly tested agents could accurately predict neomycin resistance. TAO resistance was rare in the United States after extensive OTC use and was not adversely influenced by decades of parenteral aminoglycoside use. Australian surveillance showed high levels of TAO susceptibility in sampled isolates as a baseline for possible OTC availability. TAO maintains a wider spectrum of activity compared with mupirocin and was usable against mupirocin-resistant Gram-positive strains. PMID- 16368477 TI - Etest synergy testing of clinical isolates of Staphylococcus aureus demonstrating heterogeneous resistance to vancomycin. AB - In search for potential synergistic antimicrobial combinations against Staphylococcus aureus isolates, which display heterogeneous resistance to vancomycin, we evaluated the activities of 21 various combinations involving ampicillin/sulbactam, daptomycin, gentamicin, linezolid, quinupristin/dalfopristin, rifampin, and vancomycin by Etest and time-kill methods. A number of combinations demonstrated either synergistic or additive effects against hGISA SA118 and GISA SA179. Agreement between the Etest and time kill methods for detecting antimicrobial synergy ranged from 66.6% to 71.4%. The Etest method appears promising, and further investigations are warranted. PMID- 16368478 TI - High rates of erythromycin and clindamycin resistance among OBGYN isolates of group B Streptococcus. AB - In vitro susceptibility testing on 200 Streptococcus agalactiae strains isolated during a 4-year period from vaginal/rectal specimens demonstrated a very high resistance rate for both erythromycin (54%) and clindamycin (33%). Methylase genes erm(B) and erm(TR) and efflux genes mef(E) and mef(A) were detected. Pulsed field gel electrophoresis showed evidence of both clonal spread and multiclonal dissemination of resistant strains. All but 3 of 200 isolates were susceptible to telithromycin. PMID- 16368479 TI - Manipulation and extraction of genomic DNA from cell lysate by functionalized magnetic particles for lab on a chip applications. AB - A novel approach for extracting living cells' genomic DNA materials utilizing functionalized magnetic particles (MPs) is reported in this investigation. This strategy is amenable to handle bio-samples in a miniaturized environment and it offers a possibility to separate and purify DNA from other cell lysate mixtures "on-chip", which is known to be a bottle-neck step in an integrated micro-total analysis-system (muTAS). "Species-specific" genomic DNA of interest is captured by the MPs based on the hybridization interaction between the biotinylated probes modified MPs and a complementary region of the targeted genome. The genome DNA anchored on the particles can be separated from the rest of cellular mixtures by a simple buffer washing upon the exertion of external magnetic force. Surface modifications of MPs and hybridization conditions affecting the genome capturing efficiency are investigated. Extraction of genomic DNA from E. coli is demonstrated in a silicon/glass-based micro-reactor patterned with a platinum heater and sensors. On-chip extraction and manipulation of genomic DNAs illustrated in this study is a step forward toward a total integrated bioanalytical microsystem for crude cells/sample analysis. PMID- 16368480 TI - Development and evaluation of biocompatible films of polytetrafluoroethylene polymers holding lithium phthalocyanine crystals for their use in EPR oximetry. AB - Electron paramagnetic resonance (EPR) oximetry is a powerful technology that allows the monitoring of oxygenation in tissues. The measurement of tissue oxygenation can be achieved using lithium phthalocyanine (LiPc) crystals as oxygen reporters. In order to have biocompatibility for the sensing system and to assure long-term stability in the responsiveness of the system, we developed films of Teflon AF 2400 with embedded LiPc crystals. These systems can be used as retrievable inserts or parts of an implantable resonator or catheter. Atomic force microscopy studies revealed that the surface of the films was regular and planar. The response to oxygen of the sensor (EPR linewidth as a function of pO(2)) remained unchanged after implantation in mice, and was not affected by sterilization or irradiation. The use of resonators, holding LiPc embedded in Teflon AF 2400, implanted in the gastrocnemius muscle of rabbits allowed the monitoring of oxygen during several weeks. Several assays also demonstrated the biocompatibility of the system: (1) no hemolytic effect was noted; (2) no toxicity was found using the systemic injection test of extracts; (3) histological analysis in rabbit muscle in which the films were implanted for 1 week or 3 months was similar to standard polyethylene biocompatible devices. These advanced oxygen sensors are promising tools for future pre-clinical and clinical developments of EPR oximetry. These developments can be applied for other applications of biosensors where there is a need for oxygen permeable membranes. PMID- 16368481 TI - Counter-propagating optical trapping system for size and refractive index measurement of microparticles. AB - We propose and demonstrate a novel approach to measure the size and refractive index of microparticles based on two beam optical trapping, where forward scattered light is detected to give information about the particle. The counter propagating optical trap measurement (COTM) system exploits the capability of optical traps to measure pico-Newton forces for microparticles' refractive index and size characterization. Different from the current best technique for microparticles' refractive index measurement, refractometry, a bulk technique requiring changing the fluid composition of the sample, our optical trap technique works with any transparent fluid and enables single particle analysis without the use of biological markers. A ray-optics model is used to explore the physical operation of the COTM system, predict system performance and aid system design. Experiments demonstrate the accuracy of refractive index measurement of Deltan=0.013 and size measurement of 3% of diameter with 2% standard deviation. Present performance is instrumentation limited, and a potential improvement by more than two orders of magnitude can be expected in the future. With further development in parallelism and miniaturization, the system offers advantages for cell manipulation and bioanalysis compatible with lab-on-a-chip systems. PMID- 16368482 TI - Application of europium(III) chelate-dyed nanoparticle labels in a competitive atrazine fluoroimmunoassay on an ITO waveguide. AB - We have demonstrated the use of an optical indium tin oxide (ITO) (quartz) waveguide as a new platform for immunosensors with fluorescent europium(III) chelate nanoparticle labels (Seradyn) in a competitive atrazine immunoassay. ITO as a solid surface facilitated the successful use of particulate labels in a competitive assay format. The limit of detection in the new nanoparticle assay was similar to a conventional ELISA. The effect of particle size on bioconjugate binding kinetics was studied using three sizes of bioconjugated particle labels (107, 304, and 396nm) and a rabbit IgG/anti-IgG system in a 96-well plate. A decrease in particle size resulted in faster binding but did not increase the assay sensitivity. Flux calculations based on the particle diffusivity prove that faster binding of the small particles in this study was primarily due to diffusion kinetics and not necessarily to a higher density of antibodies on the particle surface. The results suggest that ITO could make a good platform for an optical immunosensor using fluorescent nanoparticle labels in a competitive assay format for small molecule detection. However, when used in combination with fluorescent particulate labels, a highly sensitive excitation/detection system needs to be developed to fully utilize the kinetic advantage from small particle size. Different regeneration methods tested in this study showed that repeated washings with 0.1 M glycine-HCl facilitated the reuse of the ITO waveguide. PMID- 16368483 TI - Immobilisation of DNA to polymerised SU-8 photoresist. AB - SU-8 is an epoxy-based photosensitive resist, which is currently used for a large variety of MEMS and lab-on-a-chip applications. Here, we demonstrate a one-step process to functionalize SU-8 with DNA probes. The immobilisation procedure relies on direct coupling of DNA to SU-8 and resulted in surfaces with functional capture probe densities of approximately 10 fmol/mm(2) as determined by hybridisation assays with fluorescent labelled target molecules. A comparable density of functional capture probes was measured on commercial aldehyde coated glass. DNA probes did not decrease in hybridisation performance after 10 min incubation in water at 98 degrees C prior to hybridisation, indicating a covalent bond between DNA and SU-8. Finally, DNA microarrays of high quality were obtained on SU-8 by contact printing of probe solution directly on SU-8 demonstrating a simple method for the implementation of microarrays in microsystems. PMID- 16368484 TI - Endoscopic sphincterotomy permits interval laparoscopic cholecystectomy in patients with moderately severe gallstone pancreatitis. AB - Patients with moderately severe gallstone pancreatitis with substantial pancreatic and peripancreatic inflammation, but without organ failure, frequently have an open cholecystectomy to prevent recurrent pancreatitis. In these patients, prophylactic endoscopic retrograde cholangiography (ERC) with endoscopic sphincterotomy (ES) may prevent recurrent pancreatitis, permit laparoscopic cholecystectomy, and decrease risks. The medical records of all patients with pancreatitis undergoing cholecystectomy from 1999-2004 at the University of North Carolina Memorial Hospital were reviewed. Data regarding demographics, clinical course, etiology of pancreatitis, operative and endoscopic interventions, and outcome were extracted. Moderately severe gallstone-induced pancreatitis was defined as pancreatitis without organ failure but with extensive local inflammation. Thirty patients with moderately severe gallstone pancreatitis underwent ERC and ES and were discharged before cholecystectomy. Mean interval between ES and cholecystectomy was 102 +/- 17 days. Cholecystectomy was performed laparoscopically in 27 (90%) patients, open in three (10%) patients, and converted to open in two (7%) patients, with a morbidity rate of 7% (two patients). No patient required drainage of a pseudocyst or developed recurrent pancreatitis. Interval complications resulted in hospital readmission in seven (23%) patients. In conclusion, recurrent biliary pancreatitis in patients with moderately severe gallstone pancreatitis is nil after ERC and ES. Hospital discharge of these patients permits interval laparoscopic cholecystectomy, but close follow-up is necessary in these potentially ill patients. PMID- 16368485 TI - Does repeated surgery improve the prognosis of colorectal liver metastases? AB - Hepatic resection for colorectal metastases was performed for 188 patients. Overall survival rates after the first hepatectomy are 41.4% and 32.7% for 5 and 10 years, respectively. The survival rate of 116 cases with unilobar hepatic metastases (H1) is significantly higher than those of 48 cases with two to four bilobar metastases (H2) and 24 cases with more than four (H3), respectively. However, the differences between the survival rates from H1 with multiple metastases, H2, and H3 are not significant, even though the H3 group has no 10 year survivors. The 5-year survival rates after the second hepatectomy (30 patients) and the resection of the lung (26 patients) are 30.3% and 35.2%, respectively, in this series. In those patients, the 5-year survival rates from the first metastasectomy are 43.4% and 50.3%, respectively. There are 14 5-year survivors with multiple metastases and 8 of those patients underwent multiple surgeries. There are 13 patients with three or more repeat resections of the liver and/or lung. The 5-year survival rates of the patients from the first and third metastasectomy are 53.9% and 22.5%, respectively. Repeat operations for the liver and the lung contribute to the improving prognosis. PMID- 16368486 TI - Assessment of diaphragmatic stressors as risk factors for symptomatic failure of laparoscopic nissen fundoplication. AB - An important limitation of antireflux surgery is a 5%-10% failure rate. We investigated the correlation between various diaphragm stressors and failure of antireflux surgery. Forty-one study cases who underwent a reoperative antireflux operation from 1997 to 2001 and 50 control patients who had undergone a successful laparoscopic Nissen fundoplication during the same period without clinical or symptomatic evidence of failure were randomly selected for comparison. A retrospective analysis was conducted utilizing a standardized diaphragm stressor questionnaire, addressing the period between the primary and secondary operation. Stressors considered in the study included height, body mass index (BMI), postoperative gagging, vomiting, weight lifting (greater than 100 pounds), coughing, hiccuping, motion sickness, retching, belching, antidepressant use, smoking, preoperative grade of esophagitis, size of hiatal hernia, lower esophageal sphincter pressure, esophageal body pressures, and preoperative response to proton pump inhibitors. Of the potential stressors investigated, the following were significantly associated with surgical failure after adjusting for other variables through multivariate analysis: gagging (P = 0.005), belching (P = 0.02), and hernia size greater than 3 cm (P = 0.04; Table 1). Other potential risk factors show trends as obvious in Fig. 2. Vomiting was significant (P = 0.01) in the earlier models but lost significance when logistic regression was applied. Patients with postoperative gagging and an intraoperative hiatal hernia (greater than 3 cm) have a poorer outcome, whereas patients with postoperative belching have a better long-term outcome. PMID- 16368487 TI - Mapping the rectum: spatial analysis of transanal endoscopic microsurgical outcomes using GIS technology. AB - Transanal endoscopic microsurgery (TEM) is a technically challenging procedure hindered by rectal anatomic constraints. To study the relationship of lesion position with performance of TEM, a novel approach of spatial analysis using Geographic Information Systems (GIS) was developed. A retrospective review was conducted on 144 consecutive TEMs, analyzing clinical, pathologic, and positional characteristics. Two- and three-dimensional maps of rectal topology were developed. GIS was used for spatial analysis, accounting for regional position and clustering of lesions. Lesions were located at a mean distance of 9.3 +/- 4.9 (SD) cm from the dentate line, with an average size of 3.1 +/- 1.4 cm. Proximal regions were associated with prolonged operative time. Regions between the rectosigmoid junction and the peritoneal reflection were associated with peritoneal breach. In spatial regression analysis, regional characteristics that were significantly associated with operative time included distance, presence of cancers, and positive margins; peritoneal breach was significantly associated with lesion size and location; conversions were associated with distance (P < 0.05). Specific knowledge of lesion size and location in the context of anatomic relationships is important for optimizing operative intervention. GIS provides a valuable tool in organizing spatial information and can be extended into clinical research topics involving the distinction of anatomic relationships. PMID- 16368488 TI - CO2 abdominal insufflation pretreatment increases survival after a lipopolysaccharide-contaminated laparotomy. AB - Carbon dioxide (CO(2))-pneumoperitoneum is known to favorably modify the systemic immune response during laparoscopic surgery. The presented studies were designed to determine whether treating animals with CO(2) abdominal insufflation before undergoing a lipopolysaccharide (LPS)-contaminated laparotomy would serve as "shock prophylaxis" and thus improve survival and attenuate cytokine production. Rats were randomized into five groups: CO(2)-pneumoperitoneum, helium pneumoperitoneum, anesthesia control, laparotomy/LPS control, and LPS only control. Animals in the first four groups all received a laparotomy and a lethal dose of LPS. Immediately preceding their laparotomy, animals in the pneumoperitoneum groups received a 30-minute pretreatment of abdominal insufflation with either CO(2) or helium. The anesthesia control group received a 30-minute pretreatment of isoflurane. Animal mortality was then recorded during the ensuing 72 hours. Subsequently, a similar protocol was repeated for measurements of cytokines. CO(2)-pneumoperitoneum increased survival at 48 hours compared with LPS control (P <.05), and decreased interleukin-6 plasma levels at 2 hours (P <.05). Abdominal insufflation with CO(2) before the performance of a laparotomy contaminated with endotoxin increases survival and attenuates interleukin-6. The beneficial immune-modulating effects of CO(2)-pneumoperitoneum endure after abdominal insufflation. CO(2)-pneumoperitoneum pretreatment may improve outcomes among patients undergoing gastrointestinal surgery who are at high risk for abdominal fecal contamination. PMID- 16368489 TI - Comparative evaluation of gastrointestinal transit and immune response between laparoscopic and open gastrectomy in a porcine model. AB - Several reports have shown advantages of laparoscopic gastrointestinal (GI) procedures when compared with their open counterparts. The aim of this study was to assess the differences of GI transit and immune response between laparoscopic and open gastrectomy in a porcine model. Fourteen pigs were assigned to undergo partial gastrectomy, either by laparoscopy (lap group) or by laparotomy (open group). GI transit was assessed using 24 markers and measured until half of markers were evacuated. To assess immune response, we used delayed-type hypersensitivity skin antigen testing (DTH) with vaccine antigen. DTH was evaluated at 48 hours after the primary injection for induration. After 2 weeks, all animals underwent necropsy and were evaluated for adhesion formation using a scoring scale. Operation time was significantly longer in the lap group. The GI transit in the postoperative phase was significantly prolonged compared with the preoperative measured times. Postoperatively, the GI transit in the open group was significantly prolonged compared with the lap group. Immune response measured by DTH was better preserved in the lap group than in the open group. Adhesion formation was significantly less in the lap group. We concluded that laparoscopic gastrectomy resulted in faster bowel recovery and less immune suppression. PMID- 16368490 TI - Immune-enhancing diet and cytokine expression during chronic sepsis: an immune enhancing diet containing L-arginine, fish oil, and RNA fragments promotes intestinal cytokine expression during chronic sepsis in rats. AB - Chronic feeding with enteral immune-enhancing diets (IEDs) provides benefits based on composition of the diet, route of feeding, and timing of feeding in relation to timing of trauma or surgery. Our prior studies of acute feeding in naive rats demonstrated that IED promotes blood flow and proinflammatory cytokines in the ileum. We hypothesized that chronic feeding with IED would shift gut immune status to an anti-inflammatory state during chronic sepsis, resulting in an altered state of cytokine expression in the gut. Five days prior to feeding, gauze was implanted subcutaneously in the backs of male Sprague-Dawley rats, which were fed for 3 days with either control diet (CD, Boost; Mead Johnson, Evansville, IL) or IED (Impact; Novartis) and randomly assigned to one of four groups: saline control (NS) + control diet (CD), sepsis (EC) + CD, NS + IED, or EC + IED. EC rats were inoculated with 10(9) CFU Escherichia coli and 10(9) CFU Bacteroides fragilis in 2 ml normal saline into the back sponge while NS rats received 2 mL normal saline alone. After 3 days, animals were anesthetized and gut tissue samples were harvested and frozen at -80 degrees C. Tissue protein was extracted and ELISA was performed for interleukin (IL-1beta, IL-5, IL-6, IL-10, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma. In saline controls, IED feeding decreased IL-1beta, IL-5, IL-6, TNF-alpha, and IFN-gamma and increased IL-10 compared with CD-fed animals. In septic animals, IED feeding increased IL-5 and IL-6, while decreasing IFN-gamma and IL-10 in the distal third of the small intestine compared with CD-fed septic rats, whereas IL 1beta and TNF-alpha levels were unchanged. Chronic IED feeding produced a anti inflammatory state via decreased IFN-gamma and increased IL-5 and IL-6, which both promote gut IgA class switching, suggesting that the gut is shifted toward humoral immunity during chronic IED feeding in septic rats. PMID- 16368491 TI - Gastroesophageal and pharyngeal reflux detection using impedance and 24-hour pH monitoring in asymptomatic subjects: defining the normal environment. AB - Airway symptoms are often caused by aspiration of refluxed materials into the larynx. In this study we sought to define the frequency, character, and proximal extent of refluxed contents - including nonacid reflux-in normal subjects using intraluminal impedance to improve our understanding of the relationship between reflux and aspiration. Ten subjects, who had no symptoms of gastroesophageal reflux disease or airway disease, underwent impedance/pH monitoring with a catheter that allowed simultaneous esophageal and pharyngeal monitoring. Impedance detected 496 gastroesophageal reflux episodes in the 10 subjects during 240 hours of study. The majority, 399 (81% of the total) were acid reflux episodes (pH < 4). Ninety-seven were nonacid (pH > 4). Most reflux episodes (348 of 496) reached the mid esophagus (9 cm above lower esophageal sphincter). There were 51 reflux episodes that reached the pharynx (PR). Only 13 (25%) of PR were acidic (pH < 4), while 38 were nonacid. Twenty-six PR episodes were liquid and 25 were mixed (liquid and gas). The median number of PR episodes measured with impedance was 5 (0-10). In asymptomatic subjects, most episodes of gastroesophageal reflux are acidic and reach the midesophagus. Reflux into the PR appears to be more common than previously believed, and most of these episodes are not acidic. Thus, traditional 24-hour pH monitoring may underestimate the presence of pharyngeal reflux. The combination of impedance with pH monitoring markedly enhances our ability to accurately detect potential microaspiration. PMID- 16368492 TI - Outcome of 1000 liver cancer patients evaluated at the UPMC Liver Cancer Center. AB - We evaluated 1000 consecutive patients with liver tumors at the University of Pittsburgh Medical Center (UPMC) Liver Cancer Center over the 4-year period from August 2000 to August 2004. Of the 1000 patients seen, 573 had primary liver cancer and 427 had metastatic cancer to the liver. The mean age of the patients evaluated was 62.2 years, and 61% were male. Treatment consisted of a liver surgical procedure (resection or radiofrequency ablation) in 369 cases (36.9%), hepatic intra-arterial regional therapy (transarterial chemoembolization or (90)yttrium microspheres) in 524 cases (52.4%), systemic chemotherapy in 35 cases (3.5%), and palliative care in 72 patients (7.2%). For treated patients, median survival was 884 days for those undergoing resection/radiofrequency ablation, compared to 295 days with regional therapy. These data indicate that over 90% of patients with liver cancer evaluated at a tertiary referral center can be offered some form of therapy. Survival rates are superior with a liver resection or ablation procedure, which is likely consistent with selection bias. Hepatocellular carcinoma was the most common tumor seen due to referral pattern and screening of hepatitis patients at a major liver transplant center. The most common reason for offering palliative care was hepatic insufficiency usually associated with cirrhosis. PMID- 16368493 TI - Novel two-step resection for lesions between the middle hepatic vein and vena cava which allows the middle hepatic vein to be preserved. AB - Patients with bilobar colorectal cancer metastases to the liver present a unique problem in terms of resection. They sometimes require a staged approach to resection that takes advantage of the liver's ability to regenerate, as well as the newer chemotherapeutic agents (e.g., oxaloplatin, irinotecan (CPT-11), and bevacizumab) that have become available. In cases of multiple bilobar metastases, if segment IV is clear of tumor, a left lateral segmentectomy (LLS) can be performed, followed several months later by a formal right hepatectomy. The remnant liver composed of the hypertrophied segment IV is drained by the middle hepatic vein (MHV). In this context, patients with lesions between the origin of the MHV and the inferior vena cava (IVC) present a particularly difficult problem. Conventional excision would require an extended hepatectomy and division of the MHV along with either the right or left hepatic veins (RHV, LHV). This would make it impossible to continue with a formal resection of the remaining lesions in the contralateral liver without sacrificing the sole remaining hepatic vein. We present a novel two-step hepatectomy for lesions between the MHV and the IVC that allows the MHV to be preserved and all lesions to be resected. PMID- 16368494 TI - Long-term evaluation of biliary reconstruction after partial resection of segments IV and V in iatrogenic injuries. AB - Roux-en-Y hepatojejunostomy is the procedure of choice for biliary reconstruction after complex iatrogenic injury that is usually associated with vascular injuries and concomitant ischemia of the ducts. To avoid the ischemic component, our group routinely performs a high repair to assure an anastomosis in noninflamed, nonscarred, and nonischemic ducts. If the duct bifurcation is preserved, the Hepp Couinaud approach for reconstruction is an excellent choice. Partial liver resection of segments IV and V allows adequate exposure of the bile duct at its bifurcation with an anterior approach of the ducts (therefore not jeopardizing the circulation), allowing a high quality anastomosis. Long-term results of bile duct reconstruction using this approach are described. Two hundred eighty-five bile duct reconstructions were done between 1989 and 2004 in a tertiary care university hospital. The first partial-segment IV resection was done in 1994; 94 cases have been reconstructed since then using this approach. All of them had a complex injury (Strasberg E1-E5), and although in many cases the bifurcation was preserved (E1-E3), a high bilioenteric anastomosis was done to facilitate the reconstruction. In 70 cases, the bifurcation was identified, and in the 24 in which the confluence was not preserved, the right and left ducts were found except in one case. In three patients, the right duct was found unsuitable for anastomosis, and a liver resection was done. In the remaining 21, an anastomosis was done using a stent (transhepatic, transanastomotic) through the right duct. According to Lillemoe's criteria, 86 cases had good results (91%). In four of the eight remaining patients, there was the need to operate again due to the presence of an obstruction and/or cholangitis. In the rest, radiological instrumentation was done. Four of these cases have developed secondary biliary cirrhosis, two of which have died while waiting for a liver transplant, four and six years after reconstruction. Partial segments IV and V resection allows adequate exposure of the confluence and the isolated left or right hepatic ducts. Anterior exposure of the ducts allows an anastomosis in well-preserved, nonischemic, nonscarred, or noninflamed ducts. Parenchyma removal also allows the free placement of the jejunal limb, without external compression and tension, obtaining a high quality anastomosis with excellent long-term results. PMID- 16368495 TI - Alcoholic hepatitis with leukemoid reaction after surgery. AB - Alcoholic hepatitis (AH) is a clinicopathologic syndrome resulting from an excessive intake of alcohol. Leukemoid reactions (LRs) are characterized by a strikingly elevated granulocyte count over 40,000-50,000 cells/mm(3). Although a leukocytosis of 15,000-18,000 cells/mm(3) is frequently seen in AH, LRs are rare in this context. AH-associated LRs are a sign of poor prognosis and have a high mortality. A 64-year-old male with a history of heavy alcohol intake underwent a right hemicolectomy for cecal carcinoma. Preoperative laboratory data were normal with the exception of an albumin of 2.1 g/dL. Liver biopsies that were taken because of a nodular appearance revealed micronodular cirrhosis, steatohepatitis, and Mallory bodies. Postoperatively, the patient developed a leukocytosis that progressively increased to 72.6 cells/mm(3). He also developed signs of impaired hepatic and renal function. Extensive workup failed to reveal a source of infection. A trial of intravenous antibiotics had no impact on the leukocytosis. Methylprednisolone at a dose of 40 mg IV daily was started on postoperative day 9. The patient experienced a progressive decline in white blood count (WBC), which reached 25.2/mm(3) on postoperative day 14. However, he died on postoperative day 16. We conclude that the patient had AH-associated LR in the postoperative period, but died despite successful treatment of the LR with steroids. PMID- 16368496 TI - Comparison between hepatic wedge resection and anatomic resection for colorectal liver metastases. AB - Some investigators have suggested that wedge resection (WR) confers a higher incidence of positive margins and an inferior survival compared with anatomic resection (AR) of colorectal liver metastases (CLM). We sought to investigate the margin status, pattern of recurrence, and overall survival of patients with CLM treated with WR or AR. We identified 253 consecutive patients, in a multi institutional database from 1991 to 2004, who underwent either WR or AR. WR was defined as a nonanatomic resection of the CLM, and AR was defined as single or multiple resections of one or two contiguous Couinaud segments. Clinicopathologic factors were analyzed with regard to pattern of recurrence and survival. One hundred six WRs were performed in 72 patients and 194 ARs in 181 patients. There was no difference in the rate of positive surgical margin (8.3%), overall recurrence rates, or patterns of recurrence between patients treated with WR vs. AR. Patients who had a positive surgical resection margin were more likely to recur at the surgical margin regardless of whether they underwent WR or AR. The median survival was 76.6 months for WR and 80.8 months for AR, with 5-year actuarial survival rates of 61% and 60%, respectively. AR is not superior to WR in terms of tumor clearance, pattern of recurrence, or survival. WR should remain an integral component of the surgical treatment of CLM. PMID- 16368497 TI - Case-control comparison of laparoscopic versus open distal pancreatectomy. AB - Laparoscopic distal pancreatectomy is becoming an increasingly used modality in the surgical treatment of pancreatic disease. The assumption is that this will lead to shorter hospitalization and faster recovery. However, actual comparative data between open and laparoscopic distal pancreatectomy is lacking. The purpose of this study is to compare these surgical procedures. All patients who underwent either laparoscopic or open distal pancreatectomy/splenectomy were reviewed. Fifteen patients underwent laparoscopic resection, whereas 41 underwent an open resection. The 15 laparoscopic patients were matched to 15 open patients for age, gender, and pancreatic pathology. Data gathered included length of stay, pancreatic leak, postoperative complications, and return to normal activity. Of the 15 laparoscopic patients, three were converted to open operations. Laparoscopic patients had a median length of stay of 5 days (range, 3-9) compared with 8 days (range, 6-23) for the open patients (P = 0.02). The pancreatic leak rate was 13% in each group. Overall postoperative complication rate was 20% in the laparoscopic group compared with 27% in the open group. Laparoscopic patients reported a return to normal activity in 3 weeks (range, 2-7) compared with 6 weeks (range, 4-10) for open patients (P = 0.03). Laparoscopic distal pancreatectomy/splenectomy does lead to shorter hospital stay and faster return to normal activity. Pancreatic leak rate and overall complication rate appear similar. PMID- 16368498 TI - Does the literature support an indication for hepatic metastasectomy other than for colorectal primary? AB - Hepatic resection for colorectal hepatic metastatic disease has been demonstrated to have a significant outcome benefit for selected patients. Advances in anesthetic and surgical technique have resulted in a significantly reduced morbidity and mortality for this procedure, and this management approach has become widely practiced. This hepatic resection paradigm is also being applied to hepatic metastatic disease of noncolorectal origin. The purpose of this report is to review and summarize the reported literature in order to define if the current data support an indication for hepatic metastasectomy. The specific cancer primaries that this analysis evaluated include breast, melanoma, gynecologic, neuroendocrine, sarcoma, and gastric cancer. Based on the data examined, we propose that although hepatic metastasectomy for noncolorectal cancer may be a promising component of overall oncologic treatment, the role of surgical resection cannot be generalized and at present should be individualized based on the patient's clinical course and by the biologic behaviors of specific malignancies. PMID- 16368499 TI - Staphylococcal liver abscess and acute cholecystitis in a patient with Crohn's disease receiving infliximab. AB - We present an unusual case of empyema of the gallbladder associated with a pyogenic liver abscess in a patient with Crohn's disease on Infliximab. It manifested by weakness, weight loss, and vague abdominal pain, which eventually localized to the right upper quadrant 4 days prior to admission. Diagnostic evaluation, which included ultrasonography and computed tomography, revealed cholelithiasis, gallbladder wall thickening, and a low-attenuation, complex mass in the left hepatic lobe. Cholecystectomy and open drainage of the liver abscess were successfully performed. There are few reports of intrahepatic abscess associated with Crohn's disease. The relationship between acute cholecystitis and Crohn's disease has also been documented. However, this report documents the unusual complication of pyogenic liver abscess secondary to acute cholecystitis in the unique population of Crohn's disease patients on Infliximab. PMID- 16368500 TI - The value of reoperative procedures after unusual reconstructions in the gastrointestinal tract associated with substantial morbidity. AB - Reconstructive procedures of the gastrointestinal tract after resection or for bypass surgery are well established and almost completely standardized but still may cause significant morbidity. Deviations from standard reconstructive procedures have pitfalls, especially when complex reconstructions are required, and may lead to substantial morbidity. Scientific evidence for the indication to reoperate as well as the best methods to be applied is lacking and surgical experience indispensable. We report on 10 reoperative cases between 1999 and 2003 after uncommon reconstructive procedures in the gastrointestinal tract associated with substantial morbidity. In five cases (five of seven), operative correction of uncommon reconstructions in the upper gastrointestinal tract after gastrectomy, completion gastrectomy, or distal gastric resection could completely alleviate the complaints including reflux esophagitis, whereas incomplete relief of symptoms was achieved in the remaining two cases (two of seven). Corrective procedures used end-to-side esophagojejunostomy or end-to-side gastrojejunostomy with a retrocolic isoperistaltic jejunal Roux-en-Y loop and end-to-side jejunojejunostomy approximately 40 cm distal to the proximal anastomosis for biliary and exocrine pancreatic drainage. After biliodigestive anastomosis, problematic cholangitis could be completely alleviated in three cases (three of three) using end-to-side hepaticojejunostomy with a retrocolic isoperistaltic jejunal Roux-en-Y loop and end-to-side jejunojejunostomy 40 cm distal to the hepaticojejunostomy for reconstruction of the continuity of the gastrointestinal tract. Compliance with well-established standard reconstructive procedures is of elementary importance in the gastrointestinal tract. Operative correction of uncommon reconstructions associated with morbidity is usually indicated. PMID- 16368501 TI - Postoperative bezoar ileus after early enteral feeding. AB - Postoperative enteral nutrition is a widely accepted route of application for nutrition formulas due to a low complication rate, a good acceptance by patients. and a favorable cost-effectiveness. We report three cases of bezoar ileus after early postoperative enteral nutrition, using a fine needle jejunostomy (FNJ) in two cases and a nasoduodenal tube in one case. A male patient who underwent gastric resection for a gastrointestinal stroma tumor and was nourished through an fine needle jejunostomy developed an acute abdomen on the seventh postoperative day. Surgical exploration revealed a mechanical ileus caused by denaturated nutrition formula distal to the catheter tip. The second case, a female patient, underwent gastric resection for a gastric cancer and on the fourth postoperative day developed acute onset of abdominal pain. Intraoperative findings were the same as described in the first case. The third case, a male patient with necrotizing cholecystitis, underwent open cholecystectomy. Postoperative enteral feeding was performed using a nasoduodenal tube. He developed a small bowel obstruction on the 17th postoperative day that was caused by an intraluminal bezoar. In conclusion, bezoar formation represents an underestimated complication of postoperative enteral feeding. Acute onset of abdominal pain and the development of small bowel obstruction are the main clinical symptoms of this severe complication. The pathogenesis of bezoar formation remains unclear. PMID- 16368502 TI - Thrombosis of a large portal vein aneurysm: treatment by thrombectomy, aneurysmorrhaphy, and portocaval shunt. AB - An otherwise healthy 32-year-old woman had unspecific upper abdominal complaints. Diagnostic work-up, including a helical computed tomography (CT) scan and indirect splenoportography, revealed a giant extrahepatic portal vein aneurysm (PVA) extending to the central part of the splenic vein. On laparotomy, a thrombectomy and creation of a portocaval side-to-side shunt were performed. Thirteen days later, she was readmitted for re-thrombosis of the aneurysm. She underwent another laparotomy with thrombectomy and tapering of the portal venous wall (aneurysmorrhaphy) by vascular staplers. On follow-up 25 months after the operation, full relief of symptoms was noted. She was on warfarin therapy. Her portal venous system was patent. PMID- 16368503 TI - Predictive factors for pancreatic fistula after pancreaticosplenectomy for advanced gastric cancer in the upper third of the stomach. AB - This study aimed to retrospectively investigate the predictive factors for pancreatic fistula following pancreaticosplenectomy. Pancreatic fistula is a major lethal complication of pancreaticosplenectomy. However, predictive factors for this condition have not yet been established. Between April 1992 and March 2000, 147 patients with advanced gastric cancer that was located predominantly in the upper third of the stomach were enrolled in this study. Predictive factors for pancreatic fistula were investigated using univariate and multivariate analyses. Pancreatic fistula, as defined according to our criteria, was observed in 73 (49.7%) patients. In the univariate analysis, age, body mass index, serum zinc level, hyperlipidemia, and comorbid disease all significantly affected the incidence of pancreatic fistula. In the multivariate analysis, body mass index, hyperlipidemia, and comorbid disease independently predicted the occurrence of pancreatic fistula. By contrast, the experience of the operating surgeon had no significant effect on the frequency of this condition. Our results suggest that pancreaticosplenectomy, the surgical merit of which is not apparent, should be avoided whenever possible. If this operative procedure must be used (e.g., in patients with extensive tumor presence), careful manipulation and appropriate drainage are essential, particularly in cases showing predictive factors of pancreatic fistula. PMID- 16368504 TI - Differences in survival for patients with resectable versus unresectable metastases from pancreatic islet cell cancer. AB - Well-differentiated islet cell tumors can be associated with aggressive biology, resulting in early metastases to the liver. This study was carried out to determine whether survival for patients with malignant islet cell tumors and synchronous liver metastases is affected by complete surgical resection. Thirty one patients with synchronous liver metastases from islet cell cancer underwent surgical exploration with the intent for complete tumor resection, and all patients underwent resection of the pancreatic primary. The patients were divided into two groups, those with resectable versus unresectable liver metastases. Twenty-six of 31 (84%) patients underwent complete resection of both the primary tumor and all liver metastases, and 5 (16%) patients underwent only complete resection of the pancreatic primary without liver resection. To extirpate the primary tumor, a pancreaticoduodenectomy was performed in 11 of the 26 (42%) completely resected patients and in 4 of the 5 (80%) incompletely resected patients, P = NS. The remainder of the patients underwent distal pancreatectomy. There were no statistical differences in primary tumor size, lymph node metastases, or adjuvant treatments between patients with resected and unresected liver metastases. The median overall survival for the completely resected group was 78 months, longer than the 17 months for the group with unresectable liver metastases (P = 0.06). Complete tumor resection (or the tumor biology that allows such complete resection) affords a survival advantage to patients with metastatic islet cell tumors of the pancreas. Patterns of liver metastases from islet cell tumors, specifically multiple bilobar metastases that are not amenable to resection and/or ablation, predict a poor outcome despite resection of the primary pancreatic tumor. PMID- 16368505 TI - Mediastinal pancreatic pseudocyst with acute airway obstruction. AB - Pancreatic pseudocysts are usually located in the peripancreatic area, but on rare occasion a pseudocyst can reach the mediastinum. The natural history of mediastinal pseudocysts is poorly understood and seldom reported in the literature. We treated a patient who presented with an acute airway obstruction from a mediastinal pancreatic pseudocyst. Initial acute airway management and stabilization proved successful. A staged cyst decompression via a cervical and abdominal transhiatal approach was ultimately required. The natural history, potential complications, and management of pancreatic mediastinal pseudocysts are reviewed. PMID- 16368506 TI - Regulation of matrix metalloproteinase (MMP) gene expression by protein kinases. AB - Matrix metalloproteinases (MMPs) are a family of structurally and functionally related zinc-containing endopeptidases that are capable of degrading almost all of the components of the extracellular matrix (ECM). Under physiological and pathological conditions, the MMPs play a significant role in the efficient tissue turnover and remodeling. Specific MMPs are responsible for the matrix degradation and remodeling. Maintenance of the equilibrium between deposition and degradation of the extracellular matrix is essential to the normal tissue development. Therefore, synthesis and breakdown of the MMPs are tightly controlled by protein kinases which mediate a host of other cellular processes. The MMPs are often induced by several agents and any uncontrolled expression of the MMPs can contribute to the pathogenesis of many human diseases. This review focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. PMID- 16368507 TI - Using inbred mouse strains to identify genes for complex diseases. AB - In recent years, genetic studies in humans have identified a handful of genes that are associated with common disorders, but our understanding of such diseases at the genetic level remains relatively rudimentary. The use of mice to dissect the complex genetic etiology of common disorders offers a viable alternative to human studies since experimental parameters, such as environmental influences, breeding scheme, and detailed phenotyping can be controlled. This review focuses on the utility of mouse genetics for identification of complex disease genes. Atherosclerosis is used as a representative example, followed by an overview for the prospects of successful gene discovery in the future. PMID- 16368508 TI - The clinical significance of sperm-zona pellucida binding: 17 years later. AB - The development of homologous functional bio-assays for sperm quality assessment has been a focal point of reproductive biologists in order to provide a scientifically- based diagnosis in cases of fertilization failure. The availability of viable oocytes still remains an important limiting factor for laboratories to embark on the methodology of assays that examine sperm-oocyte interaction. The use of zonae pellucidae obtained from oocytes derived from post mortem tissue and failed in vitro fertilization cycles, enhanced the availability of zona material. Sperm-zona pellucida binding has been illustrated to be an essential requisite during human fertilization. This fundamental biological step can be measured under hemizona assay as well intact-oocyte test conditions. The sensitivity and specificity of sperm-zona binding results indicated the assay to be positively and significantly correlated with in vitro fertilization outcome. Furthermore, highly significant correlations were demonstrated between normal sperm morphology, hyperactivated motility, sperm creatine kinase activity and the zona binding capacity of a given sperm sample. It was concluded that andrology testing remains an ever-growing component in the work-up of the infertile couple. We enter the next millennium with many questions that remain to be answered by the hand of efficacious screening techniques and a new formidable therapy in intracytoplasmic sperm injection. PMID- 16368509 TI - Tolerance to autoimmune thyroiditis: (CD4+)CD25+ regulatory T cells influence susceptibility but do not supersede MHC class II restriction. AB - Murine experimental autoimmune thyroiditis (EAT), a model of Hashimoto's thyroiditis, has served for more than three decades as a prototypical model of T cell-mediated autoimmunity. Early investigations demonstrated a clear correlation between genetic factors, particularly the H2A locus of the MHC class II region, and susceptibility to autoimmune thyroiditis. Early studies also demonstrated that susceptibility to EAT induction could be modulated by manipulation of circulating levels of thyroglobulin (Tg), the principal thyroid antigen, resulting in the strengthening of self-tolerance. This antigen-specific induced tolerance is mediated by thymus-derived cells, and subsequent investigations revealed that the suppressive function is located in the (CD4+)CD25+ T cell subset, similar to findings in other models. We have demonstrated that these (CD4+)CD25+ regulatory T cells (Treg) influence susceptibility to thyroiditis in naive, as well as mTg-tolerized mice. Here, we describe the influence of both Treg and MHC class II haplotype, independently, as well in combination, and describe our recent utilization of MHC class II transgenic mice to directly compare the extent of their influences. PMID- 16368510 TI - The current epidemiology of cutaneous malignant melanoma. AB - As a background for understanding the increased incidence of melanoma, relevant information focuses on incidence, morality, environmental factors, host factors, and genetic factors. Incidence has increased dramatically; however, it is not clear to what extent changes in behavior, in the environment, or in early detection are involved. The major environmental factor, ultraviolet radiation exposure, shows surprisingly modest risks for developing melanoma, approximately 1.7-fold, and so focus is turning to interactions of exposure with host factors, including genetic factors. The major host factors associated with the development of melanoma include skin type and numbers of nevi (as well as atypical nevi). Genetic factors associated with familial melanoma have been well described and new attention, not yet validated, is being paid to low penetrant genes and their polymorphisms. PMID- 16368511 TI - Distinction of brain tissue, low grade and high grade glioma with time-resolved fluorescence spectroscopy. AB - Neuropathology frozen section diagnoses are difficult in part because of the small tissue samples and the paucity of adjunctive rapid intraoperative stains. This study aims to explore the use of time-resolved laser-induced fluorescence spectroscopy as a rapid adjunctive tool for the diagnosis of glioma specimens and for distinction of glioma from normal tissues intraoperatively. Ten low grade gliomas, 15 high grade gliomas without necrosis, 6 high grade gliomas with necrosis and/or radiation effect, and 14 histologically uninvolved "normal" brain specimens are spectroscopicaly analyzed and contrasted. Tissue autofluorescence was induced with a pulsed Nitrogen laser (337 nm, 1.2 ns) and the transient intensity decay profiles were recorded in the 370-500 nm spectral range with a fast digitized (0.2 ns time resolution). Spectral intensities and time-dependent parameters derived from the time-resolved spectra of each site were used for tissue characterization. A linear discriminant analysis diagnostic algorithm was used for tissue classification. Both low and high grade gliomas can be distinguished from histologically uninvolved cerebral cortex and white matter with high accuracy (above 90%). In addition, the presence or absence of treatment effect and/or necrosis can be identified in high grade gliomas. Taking advantage of tissue autofluorescence, this technique facilitates a direct and rapid investigation of surgically obtained tissue. PMID- 16368512 TI - Phosphoinositides in FCgamma receptor signaling. AB - Fcgamma receptors mediate a variety of immune responses to IgG-containing complexes. Virtually all of these responses appear to be regulated by phosphoinositides, in particular phosphatidylinositol (4,5)P2 and phosphatidylinositol (3,4,5)P3. Possible downstream effectors of phosphoinositides involved in multiple FcgammaR-mediated events are discussed, as are elements of the signaling pathways that may couple Fcgamma receptors to the enzymes of phosphoinositide metabolism. PMID- 16368513 TI - Phase II study of neoadjuvant concurrent chemioradiotherapy with oxaliplatin containing regimen in locally advanced rectal cancer. AB - The purpose of this phase II trial was to assess tolerance and efficacy of the combination of radiation, fluorouracil and oxaliplatin as neoadjuvant treatment in locally advanced rectal cancer. Between March 2001 and August 2004 , 28 patients M/18 F/10 with locally advanced rectal cancer were entered in our study. All the patients underwent to the Radiotherapy with a total dose was 45 Gy and concurrent chemotherapy with Oxaliplatin 80 mg/mq G1 on weeks 1,3,5 followed by five day continuous infusion of 5-Fluorouracile 300 mg/mq on five consecutive weeks. Surgery was planned 5 weeks later. Surgery was performed in all patients after a mean interval time of 5 weeks. Side effects and toxicity included grade II sec WHO diarrhea and grade II mucositis , grade I-II dysuria and skin reaction Downstaging to T0-2N0 was achieved in 18 patients (65%) with 4 (15%) achieving a pathologically complete response. CONCLUSIONS: Such a combined preoperative chemioradiotherapy and oxaliplatin-containing regimen is well tolerated with non increase in surgical toxicity. The good response rate observed warrants its use in further clinical trials. PMID- 16368514 TI - Surgery, radiotherapy and temozolomide in treating high-grade gliomas. AB - Temozolomide (TMZ) a recent, oral, second generation alkylating agent is a chemotherapeutic with demonstrated efficacy for the treatment of high-grade gliomas. The efficacy of TMZ has been demonstrated in both pre-clinical and phase I and II studies. The goal of this study is to determine the activity and safety of temozolomide in improving overall survival (OS), progression-free survival (PFS) and health-related quality of life (HQL) in patient with malignant gliomas treated by surgery, radiotherapy and temozolomide. Twelve patients with newly diagnosed glioblastoma (GBM), and anaplastic astrocytoma (AA) were studied. The mean follow-up period was 12 months. The overall response rate for all histological groups was 33% (4 patients), 6 patients (50%) showed a stabilization of disease. The median progression-free survival (PFS) and overall survival (OS) was respectively 8.35 and 14.1 months; time to progression was 36 week ranging from 20 to 46 In all patients, treatment with temozolomide was associated with improvement of performance status including the patient showing disease progression; Karnofski score improved in all patients by a minimum of 10, with a median of 20 at 6 months. No patient stopped the treatment due to side-effects, no major adverse events were recorded. In two cases of glioblastoma, we observed complete response and after three years, the quality of life is optimal. Surgery allows to establish a histopathological diagnosis, to improve signs and symptoms which are attributable to intracranial hypertension or tumour topography, and to reduce the number of target cells for adjunctive therapies. Radiotherapy improves survival and TMZ chemotherapy that is given after radiotherapy adds survival benefit for patients. Because of its favourable pharmacokinetic and pharmacodynamic properties and improved tolerability. Temozolomide appears to be an ideal, first-line, single-agent, with a safe profile and demonstrated HQL benefits in patients with high-grade gliomas. PMID- 16368515 TI - Stop Flow in abdominal and pelvic cancer relapses. AB - To determinate MTD, DLT and safe doses for phase II study, a dose finding study with Mitomycin and Adriamycin Stop-Flow administration was carried out. A phase II study focused on resectability of pelvic colorectal relapses is in progress. From November 1995, 84 pts, 52 male and 32 female (94 treatments), with advanced not resectable abdominal (14 pts) or pelvic (70 pts) relapses, and resistant to previous systemic chemotherapy, were enrolled in the study. 46 pts entered the phase I-early phase II study, while subsequently 38 pts were recruited in ongoing phase II study. Safe dose were: MMC 20 mg/mq and ADM 75 mg/mq. The phase II study focused on colorectal relapses registered very promising responses: 90% pain control, 1 pCR and 26 PR / 63 (OR 43%), 8 NC (13%) 9/27 responder patients (33%) obtained a complete resectability of colorectal relapses. Stop-Flow is a safe and feasible technique very useful as a palliation treatment. PMID- 16368516 TI - Matrix metalloproteinases and diseases of the central nervous system with a special emphasis on ischemic brain. AB - Matrix metalloproteinases (MMPs) are involved in the pathogenesis of several diseases of the CNS, that share common pathophysiological processes, such as blood-brain barrier (BBB) disruption, oxidative stress, remodeling of the extracellular matrix (ECM) and inflammation. In ischemic brain injury, MMPs are implicated in various stages of the disease. Early after the onset of ischemia, MMPs contribute to the disruption of the BBB leading to vasogenic edema and to the influx of leucocytes into the CNS. The ability of MMPs to digest the basal lamina of capillaries increases the risk of hemorrhagic transformation of the ischemic tissue. During the acute ischemic phase, maintenance of the ECM is essential for neuronal survival. However, ECM degradation and its reconstitution are critical to tissue recovery. MMPs as a key modulator of ECM homeostasis play a role in the cascades leading to neuronal cell death and tissue regeneration. This pleiotropic implication of MMPs in brain injury has open new areas of investigation, which should lead to innovative therapeutic strategies. Yet MMPs may have a detrimental or beneficial role depending on the stage of brain injury. Simple therapeutic strategies based on MMP inhibition have thus little chance to favorably alter prognosis. PMID- 16368517 TI - Identification and differentiation of hepatic stem cells during liver development. AB - Stem cells responsible for maintenance and repair of tissues are found in a number of organs. The liver's remarkable capacity to regenerate after hepatectomy or chemical-induced injury does not involve proliferation of stem cells. However, recent studies suggest that liver stem cells exist in both embryonic and adult livers. Using fluorescence-activated cell sorting and a culture system in which primitive hepatic progenitor cells form colonies, a novel class of cells with the marker profile c-Met(+)CD49f(+/low)c-Kit(-)CD45(-)TER119(-) was found in the developing liver. This class apparently represents the population of cells that form colonies containing distinct hepatocytes and cholangiocytes. When cells in this class are transplanted into the spleen or liver of mice subjected to liver injury, the cells migrate and differentiate into liver parenchymal cells and cholangiocytes that are morphologically and functionally indistinguishable from their native counterparts. During mid-gestation, hematopoietic cells migrate into the liver from a region bounded by aorta, gonad, and mesonephros and produce oncostatin M (OSM). In combination with glucocorticoid hormones, OSM induces maturation of liver stem and progenitor cells, including those of the c Met(+)CD49f(+/low)c-Kit(-)CD45(-)TER119(-) class. The ability to manipulate the proliferation and differentiation of liver stem cells in vitro will greatly aid in analyzing mechanisms of liver development and offers promise in stem cell therapy of liver diseases. PMID- 16368518 TI - LS-CAP: an algorithm for identifying cytogenetic aberrations in hepatocellular carcinoma using microarray data. AB - Biological techniques such as Array-Comparative genomic hybridization (CGH), fluorescent in situ hybridization (FISH) and affymetrix single nucleotide pleomorphism (SNP) array have been used to detect cytogenetic aberrations. However, on genomic scale, these techniques are labor intensive and time consuming. Comparative genomic microarray analysis (CGMA) has been used to identify cytogenetic changes in hepatocellular carcinoma (HCC) using gene expression microarray data. However, CGMA algorithm can not give precise localization of aberrations, fails to identify small cytogenetic changes, and exhibits false negatives and positives. Locally un-weighted smoothing cytogenetic aberrations prediction (LS-CAP) based on local smoothing and binomial distribution can be expected to address these problems. LS-CAP algorithm was built and used on HCC microarray profiles. Eighteen cytogenetic abnormalities were identified, among them 5 were reported previously, and 12 were proven by CGH studies. LS-CAP effectively reduced the false negatives and positives, and precisely located small fragments with cytogenetic aberrations. PMID- 16368519 TI - Experimental therapy with tissue kallikrein against cerebral ischemia. AB - Tissue kallikrein is a serine proteinase capable of cleaving kininogen substrate to produce the potent vasodilator kinin peptide. Kinin mediates a complex set of physiological actions through its receptor signaling. Systemic delivery of the kallikrein gene in an adenoviral vector significantly reduced stroke-induced mortality rate, blood pressure elevation, and aortic hypertrophy in hypertensive Dahl-salt sensitive rats fed a high salt diet. Using a focal cerebral ischemic rat model induced by middle cerebral artery occlusion, intravenous or intracerebroventricular kallikrein gene delivery significantly reduced ischemia/repefusion (I/R)-induced neurological deficits, cerebral infarction, neuronal and glial cell apoptosis, and inflammatory cell infiltration, while promoting angiogenesis and neurogenesis in the ischemic brain. A continuous infusion of a sub-depressor dose of tissue kallikrein protein through implanted minipump decreased I/R-induced neurological dysfunction and cerebral infarction, inflammation and oxidative stress independent of kallikrein's blood pressure lowering effect. Moreover, kallikrein offered neuroprotection even when delivered at one day after the onset of stroke. Kallikrein's protective effects were blocked by the kinin B2 receptor antagonist icatibant. The role of the kinin B2 receptor in mediating the protective effect against ischemic brain injury was further confirmed by increases in mortality rate and post-ischemic brain injury in kinin B2 receptor-deficient mice. Taken together, these results suggest a novel function of kallikrein as an anti-inflammatory and anti-oxidative agent in protecting the brain against ischemic stroke-induced injuries. These findings also raise the possibility that tissue kallikrein may have value in the treatment of acute ischemic stroke. PMID- 16368520 TI - Antiatherogenic role of high-density lipoproteins: insights from genetically engineered-mice. AB - Plasma levels of high-density lipoprotein (HDL) cholesterol are inversely correlated with the incidence of atherosclerotic cardiovascular disease. The cardioprotective effects of HDL have been attributed to its role in reverse cholesterol transport (RCT) and especially the macrophage-dependent RCT, and also to the antioxidant properties of HDL as well as its direct effects on endothelial function. However, few of these effects have been verified in vivo in humans. With the creation and detailed analysis of genetically-engineered mice, a solid body of new information has emerged on the mechanisms controlling these key antiatherogenic functions of HDL and their effects on atherogenesis. This article provides a review of new insights into the molecular mechanisms underlying these three most studied antiatherogenic functions of HDL in vivo with a focus on genetically-engineered mice. PMID- 16368521 TI - Tissue factor initiated blood coagulation. AB - Tissue factor (TF) is the cellular receptor and cofactor for blood coagulation factor (F) VII. Exposure of flowing blood to cells that express TF leads to the initiation of blood coagulation. Blood coagulation is tightly regulated to generate a local fibrin clot at the site of vascular injury without compromising blood flow in the vasculature. This chapter describes the initiation and propagation of the response and how it is ultimately down-regulated to prevent widespread inappropriate blood coagulation. PMID- 16368522 TI - Dendritic cells and (CD4+)CD25+ T regulatory cells: crosstalk between two professionals in immunity versus tolerance. AB - Dendritic cells (DCs) are professional antigen presenting cells (APCs). (CD4+)CD25+ T regulatory cells (T regs) are recognized as professional regulatory cells. DCs not only initiate T cell immunity by uptake, processing and presentation of specific antigens, but also induce immune tolerance by deletion of T cells and/or induction of regulatory T cells. (CD4+)CD25+ T regs maintain immune tolerance by suppressing the function of CD4+ and CD8+ T cells, B cells, macrophages, DCs and NK cells. It would be inconceivable that the delicate balance between immunity and tolerance could be kept impeccable without the crosstalk between DCs and (CD4+)CD25+ T regs. This review focuses on the recent development in our understanding of DCs and (CD4+)CD25+ T regs in immune tolerance, with transforming growth factor-beta (TGF-beta) serving as a potential link between these two professionals. PMID- 16368523 TI - Genetics of perinatal brain injury in the preterm infant. AB - Due to developmental immaturity of the central nervous system, effects of an adverse intrauterine environment and need for intensive care postnatally, preterm infants are at high risk of sustaining brain injury in the perinatal period. Infants who suffer brain injury in the perinatal period are at risk for long-term neurodevelopmental sequelae. Clinical and experimental data supports a significant role for inflammatory mediators in the pathophysiology of perinatal brain injury. Abnormalities in coagulation proteins in the sick preterm newborn may accentuate the risk for intraventricular hemorrhage. Polymorphisms in TNF alpha , IL-1 beta , IL-4, IL-6 and IL-10 as well as mutations in coagulation proteins have been investigated as potential candidate genes to modify risk and or severity of perinatal brain injury. Preliminary evidence suggests a role for cytokine genes as risk modifiers for IVH and PVL. PMID- 16368524 TI - Prostate cancer epidemiology. AB - Prostate cancer is the most common non-skin cancer among men in most western populations, and it is the second leading cause of cancer death among U.S. men. Despite its high morbidity, the etiology of prostate cancer remains largely unknown. Advancing age, race, and a family history of prostate cancer are the only established risk factors. Many putative risk factors, including androgens, diet, physical activity, sexual factors, inflammation, and obesity, have been implicated, but their roles in prostate cancer etiology remain unclear. It is estimated that as much as 42% of the risk of prostate cancer may be accounted for by genetic influences, including individual and combined effects of rare, highly penetrant genes, more common weakly penetrant genes, and genes acting in concert with each other. Numerous genetic variants in the androgen biosynthesis/metabolism, carcinogen metabolism, DNA repair, and chronic inflammation pathways, have been explored, but the results are largely inconclusive. The pathogenesis of prostate cancer likely involves interplay between environmental and genetic factors. To unravel these complex relationships, large well-designed interdisciplinary epidemiologic studies are needed. With newly available molecular tools, a new generation of large-scale multidisciplinary population-based studies is beginning to investigate gene-gene and gene-environment interactions. Results of these studies may lead to better detection, treatment, and, ultimately, prevention of prostate cancer. PMID- 16368525 TI - Microarray tools for deciphering complex diseases. AB - Individual genetic findings associated with complex diseases are unlikely to fully explain their substantial impact or provide new comprehensive insights into disease pathogenesis. These also lack the comprehensive data much needed for development of new effective drugs in majority of the disease cases in a population. In fact multilevel etiologic factors underlie almost all human diseases, including: environmental causes, epigenetic factors, DNA mutations, amplifications, and deletions, RNA expression levels, protein (translation, post translation modification, localization) and combinations thereof. Each individual might consist of different combinations of these multiple etiologic factors. Integrative evaluation of all these modifications will shed light on the whole identity of the disease and the underlying molecular mechanisms. Until now it was inconceivable to have a full grasp of such a complex etiology. Microarrays enable us to interrogate the individualized various factors (DNA, RNA and protein content) involved in disease state on genome-wide scale simultaneously and expeditiously in single cell or the tissue of interest (Figure 1). The new disciplines of microarray studies in combination hold the promise of effective, current, and comprehensive understanding of complex diseases and may be a good approach for reducing the costs and time lines associated with discovery and efficacy improvement of therapeutic drugs. In the future, through utilizing the colossal amount of microarray data findings, defining the structure, function, and dynamics of entire biological pathways and cellular networks under various physiological states, and the development of robust and efficient methods for analyzing and interpreting high dimensional data, it will be possible to connect combination of experimental results with individualized disease state. This will facilitate precise diagnosis prognosis and therapy. PMID- 16368526 TI - Different fates of donor mitochondrial DNA in bovine-rabbit and cloned bovine rabbit reconstructed embryos during preimplantation development. AB - The functions of mitochondria depend on precise interaction between nuclear and cytoplasmic genomes. Non-balance of mtDNA has been reported in most nuclear transfer embryos and offspring. The reason of the degradation of donor mtDNA is still not clear. To further investigate the mechanism, in this study, we designed an experiment as follows. Two fibroblast cell lines sharing same nuclear genome but different mitochondria genome backgrounds, namely cells from ear tissues of cloned bovine and its donor, were choose as donor cells and introduced into enucleated rabbit oocytes. Similar developmental potential was observed in cloned bovine-rabbit (clone group) and bovine-rabbit (non-clone group) embryos. Real time PCR assay showed that, in non-clone group, bovine mtDNA decreased during the development of reconstructed embryo, and that a sharp decrease was detected at the blastocyst stage. In clone group, bovine mtDNA decreased slightly, and the abrupt reduction of donor mtDNA did not occur during preimplantation development. In addition, an obvious increase in rabbit mtDNA was observed in both groups at the blastocyst stage. Our results demonstrate that: 1) the fates of donor mtDNAs in bovine-rabbit and cloned bovine-rabbit reconstructed embryos were different; and 2) recipient mtDNAs replicate at blastocyst regardless of the difference of donor cells. PMID- 16368527 TI - Pathophysiology of sperm motility. AB - Mammalian spermatozoa acquire the ability to swim during their transit from the testis to the oviduct under the control of several external and intracellular factors. These factors play also a pivotal role in regulating acquisition of hyperactivated motility and during the process of chemotaxis. This review summarizes the involvement of such factors in acquisition and maintenance of sperm motility, hyperactivation and chemotaxis, focusing in particular on the molecular bases of asthenozoospermia, a pathology of seminal plasma characterized by reduced sperm motility, which is one of the main causes of male infertility. Current in vitro treatments of asthenozoospermia are shown, together with the most recent findings on pharmacological and physiological molecules capable of stimulating sperm motility. The structure, function and mechanism of sperm flagellum responsible for the development of active motility are also analyzed in details. PMID- 16368528 TI - Signaling triggered by glucocorticoid-induced tumor necrosis factor receptor family-related gene: regulation at the interface between regulatory T cells and immune effector cells. AB - Mammals and other higher vertebrates have developed an adaptive immune system to defy effectively countless pathogens and cancerous cells encountered during the lifetime of an individual. B and T lymphocytes, which are essential in orchestrating adaptive immune responses, express surface receptors specific for foreign and abnormal self-antigens. Genesis of this antigen receptor repertoire poses significant risks for autoimmunity caused by self-reactive lymphocytes. Therefore, organisms with adaptive immune systems have evolved central and peripheral tolerance mechanisms. In peripheral tissues, regulatory T (Treg) cells function in a dominant, cell-extrinsic manner to limit inflammatory responses and autoimmune disorders. To tap the potential clinical utility of these specialized lymphocytes, advances have been made in understanding how Treg cell-mediated suppression of immune effector cells is achieved and regulated. Importantly, signaling induced by a recently identified member of the tumor necrosis factor receptor (TNFR) family, termed glucocorticoid-induced TNFR family-related gene (GITR), abrogates the suppressive effects of Treg cells. GITR plays a pivotal role in controlling T cell-mediated responses in experimental models of organ specific autoimmunity, chronic infection, and anti-tumor immunity. These findings highlight the importance of elucidating the molecular underpinnings of GITR induced signaling. We propose that GITR employs adapter proteins, including TNFR associated factors (TRAFs), to regulate diverse signaling pathways and transcriptional programs that control the interplay between Treg cells and immune effector cells. PMID- 16368529 TI - Improving the delivery of therapeutic agents to CNS neoplasms: a clinical review. AB - Even though nearly thirty-years of clinical research has attempted to improve the outcomes for patients with central nervous system neoplasms, the survival remains limited for a majority of these patients. Diverse intracellular signaling pathways involving apoptosis, invasion, angiogenesis and relevant mechanisms of resistance associated with CNS neoplasms are continuing to be elucidated. Phase I and II studies of systemically delivered chemotherapeutic agents and biological agents targeting these pathways have largely resulted in modest outcomes. Although the functional blood brain barrier was identified nearly eighty years ago only recently has the complexity and relevance of the blood brain-tumor barrier (BTB) been recognized as an important factor that limits the effective treatment of CNS neoplasms. Several groups have focused their efforts at improving the delivery of therapeutic agents across the blood brain-tumor barrier. The purpose of the article is to review novel methods that have attempted to improve the delivery of therapeutic agents into the CNS for the treatment of CNS neoplasm. PMID- 16368530 TI - Role of microtubules and myosins in Fc gamma receptor-mediated phagocytosis. AB - Microtubules serve as tracks for vesicular traffic in both phagosome formation and phagosome maturation. In the process of phagosome formation, endomembrane vesicles are delivered to the membrane of the forming phagosomes to supply membrane. This localized endomembrane delivery, referred to as focal exocytosis, facilitates pseudopod extension for the purpose of engulfing large particles. The microtubule-based transport system is the most likely candidate for such targeted vesicle trafficking to the forming phagosomes. During their maturation process, phagosomes interact with early and late endosomes and finally fuse with lysosomes. Although phagosomal membrane fusion with other membranous compartments does not require microtubules, bi-directional transport and positioning of the two organelles on microtubules are necessary for their close positioning and subsequent membrane fusion. Microtubules are also responsible for vesicle trafficking along the antigen presentation pathway for phagocytosed materials. Some classes of myosin are involved in diverse processes of Fc gamma receptor (FcgR)-mediated phagocytosis as force generators and actin-based transport motors. The role of myosin II in phagocytic cup squeezing is complementary to the classical zipper closure model. Myosin Ic and myosin X seem to be key players in extending and closing phagocytic-cup pseudopod. Other classes of myosin may also be involved in phagosomal movement. Myosin V may control short-range phagosome movement and relay phagosomes to the long-range linear transport system using microtubules. PMID- 16368531 TI - Origin and biological significance of DNA fragmentation in human spermatozoa. AB - The occurrence of DNA fragmentation in mammalian spermatozoa was identified in 1993. In human, sperm DNA fragmentation is particularly relevant in subfertile patients (i.e, those subjects more likely to be treated by assisted reproductive techniques). Thus, concerns have been raised about the possibility that sperm with DNA fragmentation may be involved in the process of fertilization, in particular when invasive techniques (such as intracytoplasmatic sperm injection) are applied. Knowledge of the mechanisms responsible for generation of DNA strand breaks may thus help in disclosing and possibly identifying new therapies for the treatment of male infertility. However, the mechanisms involved in generating sperm DNA anomalies are far from being clarified. In this review, we summarize and critically analyze the main current theories that explain generation of DNA fragmentation in spermatozoa: abortive apoptosis (anomalies in apoptosis that occur normally during spermatogenesis), problems in packaging of chromatin (mainly anomalies in histone to protamine substitution) and generation of reactive oxygen species (that may occur at any level during spermatogenesis, sperm maturation and transit in the male genital tract). PMID- 16368532 TI - Cloning, characterization and subcellular localization of a gene encoding a human Ubiquitin-conjugating enzyme (E2) homologous to the Arabidopsis thaliana UBC-16 gene product. AB - Ubiquitin charging and activation of class III E2 enzymes has been directly linked to their nuclear import. It has not been published whether other classes E2s also abide by this mechanism. During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA clone that is 2252 base pair in length, encoding a putative 162 amino acid protein, which shares high homology to Arabidopsis thaliana ubiquitin-conjugating enzyme 16 (Accession number NP_565110, 51% identity and 71% similarity) at protein level. Bioinformatics analysis revealed that the gene is composed of 7 exons, located on human chromosome 8q13-8q21.1, and that the predicted protein of the gene is a class I E2, for only composed of a conserved approximately 150-amino acid catalytic core, ubiquitin-conjugating enzyme E2 domain (UBC domain). In the C-terminal of the UBC domain sequence, there are two nuclear localization signals (NLSs). RT-PCR showed that this gene is ubiquitously expressed in 16 kinds of normal human tissues, but expression level is very low, unless in human heart, brain, liver, and pancreas. The subcellular localizations of the new human Ubiquitin conjugating enzyme E2 and its mutation were also examined, which showed that the nuclear localization of hUBC16 depended on two conditions: It has NLS, and at the same time, has enzyme active site, too, at least in HEK293 cells. PMID- 16368533 TI - Posttranslational modifications of Bcl2 family members--a potential therapeutic target for human malignancy. AB - Apoptosis is a process that can occur normally, such as during tissue remodeling, embryogenesis or abnormally during certain pathologies, such as cancer. The identification of the Bcl2 as well as IAP family members has suggested that excessive inhibition of apoptosis may constitute a common feature of all known human cancers-the ability to influence their onset, progression and outcome. Bcl2 family proteins are frequently regulated by phosphorylation that affects their activity and conformation. The structural analysis of antiapoptotic members of Bcl2 family has contributed to a better understanding of the functional domains including the discovery of an unstructured "loop region" (LR) near the N-terminus exposed to the cytoplasm. The antiapoptotic members of Bcl2 family such as Bcl2/Bcl-xL/Mcl-1 are phosphorylated on specific serine/threonine residues within this unstructured loop in response to diverse stimuli including treatment with chemotherapeutic taxanes, survival factor addition or chemopreventive agents. In most instances, such phosphorylation has been associated with the loss of their biological function. The chemoresistant tumors overexpress Bcl2/Bcl-xL/Mcl-1. To this end, the apoptosis yielding effect due to phosphorylation of antiapoptotic Bcl2 family members is quite interesting. Phosphorylation-dephosphorylation pathway of these antiapoptotic proteins should be an ideal molecular target for therapy of subpopulation of cancer in which these death repressors are essential prognostic markers. Thus, further gaining the knowledge on the mechanism of inactivation of Bcl2/Bcl-xL/Mcl-1 by phosphorylation might be of paramount importance to therapy for human malignancies in which overexpression of these antiapoptotic proteins plays an essential role. PMID- 16368534 TI - Regulation of protein 4.1R interactions with membrane proteins by Ca2+ and calmodulin. AB - Red blood cell protein 4.1 (4.1R) is essential for maintaining erythrocyte shape and controlling membrane mechanical properties, such as deformability and stability. The importance of 4.1R has been demonstrated by the dramatic erythrocyte alterations observed in patients lacking this protein. Indeed, 4.1R null red blood cells adopt an elliptical shape and are characterized by unstable membranes. The key role of 4.1R likely results from multiple protein-protein interactions: lateral interactions with the spectrin/actin network and vertical interactions with the cytoplasmic domain of transmembrane proteins glycophorin C (GPC), Band 3 (anion exchanger 1, AE1), and CD44. 4.1R promotes the formation of a ternary complex with GPC and p55 through its 30 kDa membrane-binding domain. Based on the primary structure of the prototypical 80 kDa isoform of 4.1R, functional domains and sites for binding partners have been identified. The others and we have been focusing on the structure and function of the 30 kDa NH2 terminal domain of 4.1R, which is responsible for 4.1R interaction with the transmembrane proteins described above. A major finding is that Ca2+, in association with calmodulin (CaM), plays a critical role in regulation of the interaction of the 30 kDa domain with its various binding partners. This review is a detailed report of our current knowledge regarding 4.1R, and more specifically, 4.1R 30 kDa domain: its primary structure, functions and modulation by Ca2+ and CaM. Emphasis is given on the relationships between structure and function that we have been able to establish through X-ray crystal structure analysis of the 30 kDa membrane-binding domain in 4.1R. Finally, we give insights into the potential roles of 4.1R in the dynamic organization of the membrane skeleton viewed as a complex system. PMID- 16368535 TI - Alteration of Rb binding to HPV 18 E7 modified by transglutaminase 2 with different type of polyamines. AB - High-risk human papillomavirus (HPV) E7 is a major oncoprotein that plays a crucial role in the development of cervical cancer. A previous study showed that transglutaminase (TGase) 2 catalyzes the incorporation of polyamines into HPV 18 E7 protein, and thereby diminishes its ability to bind Rb. Therefore, TGase 2 activity may be implicated in a suppressive function of host against HPV-induced carcinogenesis. To better understand the nature of polyamination of HPV 18 E7, we investigated the Rb binding of E7 polyaminated in vitro with different type of polyamines. The incorporation of spermine diminished the Rb binding of E7 more profoundly compared with that of spermidine, suggesting that either the additional positive charge or a steric effect or both may have altered the chemical or structural properties of the protein. In addition, the treatment of either spermidine or spermine in cultured cell system reduced the ability of E7 to inactivate Rb with a TGase activity-dependent manner. Spermine was more effective in inhibiting E7 activity than spermidine. These results may provide the basis for future investigation aiming at delineating the significance of polyamine metabolism on HPV E7 functions. PMID- 16368536 TI - APO2 ligand/tumor necrosis factor-related apoptosis-inducing ligand in prostate cancer therapy. AB - Prostate cancer is one of the most common cancers in men and is the second leading cause of cancer-related death in the USA. Many anti-tumor agents against prostate cancer cells have been developed, but their unacceptable systemic toxicity to normal tissues usually limits their use in the clinic. Apo2 ligand (Apo2L), also called Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), is one of several members of the TNF gene superfamily that induces apoptosis through engagement of death receptors. This protein has generated tremendous enthusiasm as a potential tumor-specific cancer therapeutic because, as a stable trimer, it selectively induces apoptosis in many transformed cells, but not in most normal cells. In this review we discuss its potential use in prostate cancer therapy, the mechanisms by which induces apoptosis or that underlie resistance to it, and strategies for sensitization to overcome them. Conventional chemotherapeutic and chemopreventive drugs, irradiation, and other therapeutic agents, such as histone deacetylase inhibitors or retinoids can sensitize Apo2L/TRAIL-resistant cells and tumors. Investigating the apoptotic effects of Apo2L/TRAIL, a unique tumor-specific cell death ligand, now in clinical trials, alone or in combination may not only help in understanding its antineoplastic role in prostate carcinoma but may also provide insights into basic mechanisms of apoptosis. PMID- 16368537 TI - Neutrophil depletion inhibits early and late monocyte/macrophage increase in lung inflammation. AB - Monocytes/macrophages have critical impact on outcomes of lung inflammation. Kinetics and mechanisms for the increase of monocytes/macrophages in lungs are not completely understood. To better understand these mechanisms, E. coli-LPS (250 micro grams; N = 35) or endotoxin-free saline (N = 5) were instilled intratracheally in Sprague-Dawley rats and the increase in monocytes/macrophages, neutrophils and monocyte chemotactic protein-1 (MCP-1) was quantified at various time points after LPS treatment. In contrast to typical pattern of neutrophil influx between 6 and 24 hours, monocytes/macrophages increased in two distinct phases, very early at 3 hours and late at 24 hours. The role of neutrophils in monocyte/macrophage increase was addressed in LPS-challenged neutropenic rats (N = 8). Neutrophil depletion before instillation of LPS abrogated the early as well as late monocyte/macrophage increases in the lung. Quantification of MCP-1, which is one of the major chemoattractants for monocytes, in lung homogenates showed similar concentrations in neutropenic and non-neutropenic LPS-challenged rats. These findings show that increase in monocytes/macrophages in lung occurs in two, early and late phases, both being dependent on neutrophils but not on MCP-1. PMID- 16368538 TI - The promoter competition assay (PCA): a new approach to identify motifs involved in the transcriptional activity of reporter genes. AB - Identifying particular motifs responsible for promoter activity is a crucial step toward the development of new gene-based preventive and therapeutic strategies. However, to date, experimental methods to study promoter activity remain limited. We present in this report a promoter competition assay designed to identify, within a given promoter region, motifs critical for its activity. This assay consists in co-transfecting the promoter to be analyzed and double-stranded oligonucleotides which will compete for the binding of transcription factors. Using the recently characterized SBEM promoter as model, we first delineated the feasibility of the method and optimized the experimental conditions. We then identified, within an 87-bp region responsible for a strong expression of the reporter gene, an octamer-binding site essential for its transcriptional regulation. The importance of this motif has been confirmed by site-directed mutagenesis. The promoter competition assay appears to be a fast and efficient approach to identify, within a given promoter sequence, sites critical for its activity. PMID- 16368539 TI - The prognostic role of pre-chemotherapy hemoglobin level in patients with ovarian cancer. AB - Anemia significantly affects quality of life of cancer patients, but the impact of hemoglobin levels on survival is still unclear. The aim of this retrospective study was to assess the prognostic role of pre-chemotherapy hemoglobin levels in patients with ovarian cancer. Two hundred twenty-two patients were divided in 3 groups based on baseline hemoglobin levels (< 10 gr/dl (54 pts., 24%); 10-11.9 gr/dl (87 pts., 39%), > or = 12 gr/dl (82 pts., 37%)). Correlations among baseline characteristics (age, ECOG performance status, stage, grading, histology, residual disease after primary surgery) and baseline hemoglobin level were analyzed. Poor performance status (p = 0.03), more advanced stage (p = 0.01), and sub-optimal residual disease (p = 0.002) were more frequent in patients with lower hemoglobin values. There was no significant correlation between baseline hemoglobin level and response rate to subsequent chemotherapy. Based on univariate analysis, hemoglobin categories were statistically significant predictors for time to progression (p = 0.0002) and overall survival (p < 0.0001). Based on multivariate analysis, patients with hemoglobin between 10 and 12 g/dl had a 1.45 hazard ratio (HR) for recurrence and a 1.35 HR of death compared with patients with normal hemoglobin. Patients with hemoglobin < 10 g/dl had a 2.02 HR of recurrence and a 2.49 HR of death compared with patients with normal hemoglobin. These findings show that hemoglobin level prior to chemotherapy is an independent predictor of progression-free survival and overall survival in patients treated for ovarian carcinoma. PMID- 16368540 TI - Transglutaminases in mineralized tissues. AB - Bone development and formation during embryogenesis as well as postnatally during bone remodeling is a complex process controlled systemically and locally by hormones, growth factors and matrix molecules. Transglutaminases (TGases) are the protein cross-linking enzymes, which have long been implicated in bone development and formation. Two members of TGase family, TG2 (also called tissue transglutaminase) and FXIIIA (the enzymatic A subunit of coagulation factor XIII), are expressed in chondrocytes and osteoblasts. The results of analyses in vivo and in vitro accumulated to date indicate an important role of these enzymes in promoting chondrocyte and osteoblast differentiation and matrix mineralization. These effects could be mediated by protein cross-linking activity of TGases, by GTPase activity of TG2 or via non-catalytic signaling effects. The aim of this review is to summarize the available data regarding the expression, localization and activity of TG2 and FXIIIA in mineralizing tissues and to discuss a number of mechanisms by which TGases could exert their promineralizing effects. PMID- 16368541 TI - Insights into transcriptional regulation by FOXP3. AB - FoxP3 recently entered the spotlight as a critical component of regulatory T cell development and function. Several groups are presently engaged in an effort to uncover the mechanistic details of its contribution to this critical T cell subset. Despite this, the mechanism of FoxP3-mediated transcriptional repression and the affected target genes are still largely unknown. First, we discuss insights from work on other Fox family members with an emphasis on those with known roles in the immune system. Second, we review recent data concerning the molecular mechanism of FoxP3 function and its role in human disease. Finally, we consider what is known about FoxP3 target genes and their effect on T cell physiology. PMID- 16368542 TI - Monoclonal antibody therapy. AB - The concept of targeted therapy was conceived through increased understanding of the biological pathways involved in the pathogenesis of cancer and subsequently identification of the most appropriate antigens to target. Monoclonal antibody therapy harnesses host defense mechanisms through activation of the antibody dependent cytotoxic pathway and complement mediated cytotoxicity. However, these two processes alone do not explain the therapeutic efficacy of antibody therapy; they also act by apoptotic signaling and growth inhibitory pathways. Conjugation of monoclonal antibody therapy, with radionuclides or toxins, offers more therapeutic approaches. Initial data demonstrates efficacy of single agent use, although combination therapy appears potentially more beneficial. Monoclonal antibody therapy is having a significant impact on many disease processes, particularly malignancies of solid and hematological origin. In this article, we shall review and discuss the monoclonal antibodies approved by the US Food and Drug Administration (FDA). in the management of cancer. PMID- 16368543 TI - Regulation of mammalian sperm capacitation by endogenous molecules. AB - Capacitation in vitro in mammalian spermatozoa can be regulated by a number of first messengers, including fertilization promoting peptide, adenosine, calcitonin and angiotensin II, all of which are found in seminal plasma. The responses appear to involve several separate signal transduction pathways that have a common end point. These seminal-plasma derived first messengers can bind to specific receptors and directly or indirectly modulate the activity of membrane-associated adenylyl cyclase isoforms and production of the second messenger cAMP. Responses to all of these except angiotensin II involve initial acceleration of cAMP production and capacitation followed by inhibition of both cAMP production and spontaneous acrosome loss, resulting in maintenance of fertilizing potential. Appropriate G proteins and various phosphodiesterase isoforms also appear to be involved. The transition from stimulatory to inhibitory responses involves loss of decapacitation factors (DF) from receptors (DF-R) on the external surface; a DF-R present on both mouse and human spermatozoa has recently been identified as phosphatidylethanolamine-binding protein 1. The presence/absence of DF appears to cause changes in the plasma membrane that then alter the functionality of various membrane-associated proteins, including receptors. Since spermatozoa contact these first messengers at ejaculation, it is plausible that their actions observed in vitro also occur in vivo, allowing these molecules to play a pivotal role in enhancing the chances of successful fertilization. PMID- 16368544 TI - New insights into potential functions for the protein 4.1 superfamily of proteins in kidney epithelium. AB - Members of the protein 4.1 family of adapter proteins are expressed in a broad panel of tissues including various epithelia where they likely play an important role in maintenance of cell architecture and polarity and in control of cell proliferation. We have recently characterized the structure and distribution of three members of the protein 4.1 family, 4.1B, 4.1R and 4.1N, in mouse kidney. We describe here binding partners for renal 4.1 proteins, identified through the screening of a rat kidney yeast two-hybrid system cDNA library. The identification of putative protein 4.1-based complexes enables us to envision potential functions for 4.1 proteins in kidney: organization of signaling complexes, response to osmotic stress, protein trafficking, and control of cell proliferation. We discuss the relevance of these protein 4.1-based interactions in kidney physio-pathology in the context of their previously identified functions in other cells and tissues. Specifically, we will focus on renal 4.1 protein interactions with beta amyloid precursor protein (beta-APP), 14-3-3 proteins, and the cell swelling-activated chloride channel pICln. We also discuss the functional relevance of another member of the protein 4.1 superfamily, ezrin, in kidney physio-pathology. PMID- 16368545 TI - Regulation of matrix metalloproteinase-13 and tissue inhibitor of matrix metalloproteinase-1 gene expression by WNT3A and bone morphogenetic protein-2 in osteoblastic differentiation. AB - During bone remodeling, degradation of skeletal connective tissue is regulated, at least in part, by the balance between matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinase (TIMPs), their natural inhibitors. Recently, the Wnt signaling pathway has been demonstrated to play a crucial role in the regulation of bone formation. Here, we investigated a potential role for Wnt signaling and functional cross-talk with bone morphogenetic protein (BMP)-2 in mRNA expression of MMPs, TIMPs and bone matrix proteins in pluripotent C2C12 cells. To assess the functional contribution of Wnt signaling, we have generated C2C12 cell lines stably over-expressing Wnt3a or Wnt5a, and then treated these cells with BMP-2 for 24 h. In these cultures, MMP-13 mRNA expression was induced by BMP-2 in Wnt3a over-expressing C2C12 (Wnt3a-C2C12) cells but not in either Wnt5a over-expressing C2C12 (Wnt5a-C2C12) cells or vehicle-transfected C2C12 cells. MMP-13 mRNA was induced in these cells by addition of BMP-2 for 12 h and the enhancement lasted up to 48 h. These effects were observed in a dose dependent manner. Enzymatic activity of MMP-13 also induced in Wnt3a-C2C12 cells by addition of BMP-2. However, membrane type-1 matrix metalloproteinase (MT1-MMP) and MMP-2 mRNA expression was not affected by either Wnt3a or BMP-2. In contrast, TIMP-1 mRNA expression was suppressed by BMP-2 in Wnt3a-C2C12 cells but not in Wnt5a-C2C12 cells. Our results show that expression of MMP-13 and TIMP-1 is regulated by Wnt signaling combined with BMP-2 in osteoblastic differentiation, and this signaling may in part mediate MMP-13 and TIMP-1 production during bone formation and/or remodeling. PMID- 16368546 TI - A sperm component, HSD-3.8 (SPAG1), interacts with G-protein beta 1 subunit and activates extracellular signal-regulated kinases (ERK). AB - HSD-3.8 cDNA (accession number AF311312) encodes a human sperm component. A 0.7 kb fragment (HSD-0.7) containing three immunological epitopes of HSD-3.8 cDNA was prepared and expressed in E. coli. Immunization of female rats with the recombinant HSD-0.7 proteins induced infertility. A cDNA fragment encoding the C terminal 144 amino acids of human G-protein beta l subunit (Gbeta1-C144) was screened by yeast two-hybrid, when HSD-0.7 segment was used as a bait. Recombinant His6-tagged-Gbeta1-C144 protein was expressed in E. coli BL21 and Anti-Gbeta1 serum was raised with purified Gbeta1-C144. HA-tagged HSD-0.7 and FLAG-tagged Gbeta1 plasmids were constructed and co-transfected into human embryonal kidney 293 cells. Two proteins were localized at superimposable sites in the cytoplasm, and they formed a complex when 500 micromol/L GDP existed. Overexpression of HSD-0.7 activated the G-protein-mediated extracellular signal regulated kinases (ERK1/2); however, the truncated fragments of HSD-0.7, which lacked either TPR domain or P-loop, lost the ability to activate the ERK1/2 pathway. Further study revealed that the activation of ERK1/2 was protein kinase C (PKC) rather than Ras dependent. These results provide evidence that HSD-3.8 present in spermatocytes and sperm may participate in spermatogenesis and fertilization process by activating the PKC-dependent ERK1/2 signal transduction pathway. PMID- 16368547 TI - Hydrostatic pressure increases apoptosis in cartilage-constructs produced from human osteoarthritic chondrocytes. AB - Tissue-engineering is considered a promising avenue for developing human articular cartilage implants that can be employed for resurfacing damaged cartilage in the early stages of osteoarthritis. In the present study, human cartilage-constructs were produced from human osteoarthritic chondrocytes maintained on a scaffold of HYAFFR-11 in perfusion mini-bioreactors or after implantation and recovery from nude or SCID mice after 3 weeks. The human cartilage-construct extracellular matrix reacted positively with anti-Type II collagen monoclonal antibody, but not with anti-Type I or anti-Type X collagen monoclonal antibodies. A significant portion of the cartilage-construct extracellular matrix stained metachromatic with Toluidine blue-O indicative of sulfated-proteoglycan deposition. Cyclic hydrostatic pressure applied for 4 hrs at 5 MPa using a 1 Hertz sinusoidal frequency significantly increased (p < 0.02) the proportion of apoptotic cells in the cartilage-constructs (41% +/- 4.2%; mean +/- SD) compared to control cartilage-constructs (28.5 +/- 8.4%). PMID- 16368548 TI - Matrix metalloproteinases (MMPs) in health and disease: an overview. AB - Matrix metalloproteinases (MMPs) are members of an enzyme family that require a zinc ion in their active site for catalytic activity. MMPs are critical for maintaining tissue allostasis. MMPs are active at neutral pH and can therefore catalyze the normal turnover of extracellular matrix (ECM) macromolecules such as the interstitial and basement membrane collagens, proteoglycans such as aggrecan, decorin, biglycan, fibromodulin and versican as well as accessory ECM proteins such as fibronectin. Members of the MMP family include the "classical" MMPs, the membrane-bound MMPs (MT-MMPs) the ADAMs (a disintegrin and metalloproteinase; adamlysins) and the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motif). There are more than 20 members in the MMP and ADAMTS family including the collagenases, gelatinases, stromelysins, some elastases and aggrecanases. Adamlysins are membrane-bound MMPs that also degrade aggrecan, but more importantly, one ADAM family member (i.e.ADAM-17) is a tumor necrosis factor alpha (TNF-alpha)-converting enzyme (TACE) that activates pro-TNF-alpha. Most of the MMPs are synthesized as inactive latent enzymes. Conversion to the active enzyme is generally mediated by activator systems that include plasminogen activator or the pro-hormone convertase, furin. MMP activity is regulated by a group of endogenous proteins, called, tissue inhibitor of metalloproteinases (TIMPs) that bind to active and alternative sites of the activated MMP. Significant advances have occurred in the understanding of the regulation of MMPs, ADAMs and ADAMTSs gene expression. In addition, development of MMP inhibitors to study MMP structure/function relationships spawned many studies to determine the effectiveness of MMP inhibitors in regulating abnormal connective tissue turnover. In addition, development of MMP null mice carrying specific MMP deletions has provided an opportunity to explore the role of MMPs in normal development as well as in such diverse conditions and diseases as skeletal dysplasias, coronary artery and heart disease, arthritis, cancer, and brain disorders. PMID- 16368549 TI - Matrix metalloproteinases: role in skeletal development and growth plate disorders. AB - Differentiation is the cellular process that regulates development of long bones and joint surface cartilage of synovial cavities. Growth plate cartilage development is commonly referred to as endochondral ossification which is the end stage of long bone development. Endochondral ossification proceeds as a continuum of chondrocyte proliferation cycles followed by non-proliferative phases coupled to extracellular matrix protein transformations that are regulated by proteins of the hedgehog family and by parathyroid-hormone-related peptide and its receptor, the parathyroid-hormone-related peptide receptor. A compelling body of evidence has now emerged implicating matrix metalloproteinases (MMPs) in the process of long bone lengthening and endochondral ossification. Among the MMPs, MMP-13 (collagenase-3), MMP-9 (92-kDa gelatinase; gelatinase B) and MMP-14 (MT1-MMP) are the most abundant proteinases that regulate cellular migration, alterations in the extracellular matrix and apoptosis in growth plate cartilage. Murine mutation or ablation models of growth plate development that target MMPs often result in skeletal abnormalities, indicating the critical role that MMPs play in these animal models and in skeletal maturation. Many of the MMPs that have been identified as regulating the spatial and temporal changes in rodent and rabbit endochondral ossification have also been identified by in situ hybridization and immunohistochemical analysis of human long bone development. Genetic manipulation to correct defective or dysfunctional MMP genes or MMP activity found in certain human chondrodysplasias may provide a novel strategy for treating medical disorders characterized by skeletal anomalies. PMID- 16368550 TI - TAT fusion proteins against ischemic stroke: current status and future perspectives. AB - The delivery of proteins across the blood-brain barrier is severely limited by the proteins' size and biochemical properties. Thus, a large number of peptides have been characterized in recent years that efficaciously prevent neuronal death in vitro, but which may not be applied in vivo, since they are unable to cross cell membrane barriers. In the 1990ies, it had been shown that the HIV TAT protein is able to cross cell membranes even when coupled with larger peptides. Subsequent studies with fusion proteins of the 11-amino acid protein transduction domain of HIV-TAT revealed that TAT fusion proteins may successfully be used for therapeutic purposes in vivo, even when systemically applied. Indeed, intravenous delivery of TAT proteins linked with anti-apoptotic (Bcl-XL) and neurotrophic (GDNF) factors resulted in a rapid and highly efficacious transduction of the brain tissue. When administered after focal cerebral ischemia, intravenous TAT Bcl-XL and TAT-GDNF significantly reduced brain injury, both when applied after severe and mild ischemic insults. These data provided the first in vivo evidence of the efficacy of fusion proteins in the ischemic brain, thus raising new hopes that neuroprotection is feasible after stroke. Yet, molecular biological studies have pointed out more recently that there are also limitations of the TAT protein strategy, which still need to be addressed. The development of clinically applicable treatments for ischemic stroke based on fusion protein technologies deserves concerted efforts in the future. PMID- 16368551 TI - A clinoptilolite effect on cell media and the consequent effects on tumor cells in vitro. AB - Clinoptilolite is a nontoxic natural zeolite with properties of an ion-exchanger and adsorbent. Earlier studies showed that clinoptilolite could be an adjuvant in cancer therapy. The aim of this study was to define effects of clinoptilolite in cell media on cell viability and activity of key proteins regulating cell survival, cell division and stress response. The number of viable cells, DNA synthesis and activity of EGF-R, PKB/Akt and NF?B was reduced, while apoptosis was increased in cells that were cultured in medium supplemneted with clinoptilolite. These results might be due to adsorbtion of some serum components such as EGF to clinoptilolite. In treated medium without serum the predominant role of clinoptilolite is that of cation exchange, likely affecting calcium levels and calcium-dependent signalling pathways. These results are in line with other data that confirm enhanced apoptosis in cells incubated in treated medium. Together, data presented here demonstrate that clinoptilolite affects cellular microenvironment through mechanisms that are dependent on adsorptive and ion exchange characteristics of this material. PMID- 16368552 TI - Optimal design of Ig 5' primers for construction of diverse phage antibody library established to select anti-HAb18GEF and anti-DOTA-Y Fabs for hepatoma pretargeting RIT. AB - Phage antibody library yields antibodies with higher affinity against different antigens, if diverse IgV gene repertoires can be amplified. As the currently available Fab primer sets cannot guarantee efficient amplification with high diversity, and because rare cloning sites can be found in certain Ig genes, here, we present an optimal set of Ig 5' primers, compatible with Fd 5' clone site replaced pComb3 vector, for diverse Fab phage display library construction. These novel Fab primes designed based on the newly classified IgV families, not only have best match and highest coverage for IgV family with minimized N-terminal amino acid changes, but also present good amplification diversity and efficiency of Ig gene from mice immunized with different forms of antigens (HAb18GEF, KLH DOTA-Y, and HAb18G/pcDNA3). A high quality immune phage library with good diversity was constructed based on the mixed Ig repertoire, and five high affinity Fab antibodies were selected to specifically bind to HAb18GEF, DOTA-Y and an irrelevant antigen gamma-sm, respectively. This novel Fab primers set can be applied to the construction of diverse phage antibody library and the anti HAb18GEF and anti-DOTA-Y Fab antibodies lay a solid foundation for radioimmunotherapy of hepatoma. PMID- 16368553 TI - Inflammation and stroke: therapeutic effects of adenoviral expression of secretory Leukocyte Protease Inhibitor. AB - Inflammation is a fundamental reaction of the organism aimed to protect, defend and amend diverse forms of injuries. Stroke, an ischemic condition of the brain, provokes acute and chronic inflammatory reactions composed of brain cells as well as circulating immune inflammatory cells. In response to ischemia, brain cells transcribe and express inflammatory proteins that orchestrate numerous biochemical pathways, some of which of potential detrimental effects. This review paper is aimed to delineate strategies that may enable the discovery of novel therapeutic targets for the discovery of drugs for stroke. Soluble Leukocyte Protease Inhibitor (SLPI) serves as a prime example for this strategy. PMID- 16368554 TI - The role of transglutaminase-2 and its substrates in human diseases. AB - The most characteristic enzymatic function of the class of enzymes known as transglutaminases (TG, EC 2.3.2.13) is the formation of covalent bonds between epsilon-amino groups of primary amines (from lysines or others) and the gamma carboxamine group of glutamine residues of proteins. In the last years, a growing body of evidence indicate that the most interesting member of the TG family, namely the tissue TG (tTG, also called transglutaminase type 2, TG2), possesses more than one catalytic function. In fact, TG2 is able to catalyze a crosslinking reaction, a deamidation reaction and also shows GTP-binding/hydrolyzing and isopeptidase activities. Therefore, it can act on several classes of substrates, ranging from proteins to peptides, small reactive molecules like mono- and polyamines, and nucleotides. Given the broad spectrum of potentially different activities, elucidating the role of TG2 and its substrates in cellular functions and human diseases is a difficult task. In this study we focus our attention on substrates of TG2 and report a number of interesting considerations about their possible interplay in biological processes and involvement in human diseases, including genetic disorders. A significant improvement in understanding this complex scenario may come from a "multi-interfaced" approach, by exploiting different bioinformatic tools. Starting from a database of known TG2 substrates and using bioinformatic cross-search among other databases, we generated relational tables from which an involvement of TG2 in several genetic disorders can be hypothesized. Developing new bioinformatic tools and strategies to investigate the role of TG2 in molecular mechanisms underlying human diseases will add new light to this fascinating field of research. PMID- 16368555 TI - Gene chromosomal organization and expression in cultured human neurons exposed to cocaine and HIV-1 proteins gp120 and tat: drug abuse and NeuroAIDS. AB - As a model for Neuropsychiatric dysfunction in NeuroAIDS due to HIV-1 infection and drug abuse, we analyzed gene expression in human neurons treated with cocaine and HIV-1 proteins tat and envelope (env). One-way ANOVA showed statistically significant genes among the treatment groups (p < or = 0.0005). The identified genes were then subjected to a "stepwise" analysis using a repeated measures ANOVA to discover genes with parallel response group profiles across the treatment conditions. These groups were then analyzed using a repeated measures ANOVA to assess treatment main effects and gene-by-treatment interactions within groups. One-way ANOVA produced 35 genes that were significantly associated across all treatment conditions. Factorial analysis of each gene found statistically significant differences: 30--tat, 17--cocaine, 10--env, 6--tat/env, 6--coc/env, and 4--coc/tat. Analyses across genes found three sets of four genes, one set of three genes, and three sets of two genes with parallel profiles. Identified genes had functions included signaling, immune related, and transcription control. The genes were not stochastically arranged on the chromosomes, were in proximity to each other, and to other genes involved in neuropsychiatric diseases. We hypothesize that these genes fall in transcriptionally isolated groups and that abused drugs and HIV-1 proteins trigger transcription overload, coerced expression that may result in damage to the chromosome's control and organization of chromatin transcription machinery. PMID- 16368556 TI - Stress-induced plasticity of monoamine axons. AB - Central serotonergic (5-HT) and noradrenergic (NA) neurons, which innervate the same regions of the brain, are known to play a crucial role in emotion and mood. These monoamine neurons have a great capacity to alter axonal morphology in response to repeated stress. The morphological responses of 5-HT and NA axons to repeated stress are different, and they sometimes even demonstrate opposite responses (namely, either sprouting or degeneration). Moreover, a morphological interaction also occurs between 5-HT and NA axons during axonal regeneration. This review describes the differential features of axonal plasticity of 5-HT and NA neurons in relation to stress, and discusses the possible roles that the morphological plasticity of 5-HT and NA axons may play in the pathophysiology of depression. PMID- 16368557 TI - Microarray technology in the investigation of diseases of myocardium with special reference to infection. AB - Microarrays are now routinely employed to characterize gene expression of thousands of genes from a single hybridization. The genome wide gene expression profile aids in the understanding of genes that may be regulated in a particular pathological condition. This paper provides an overview of microarray technology and its recent developments followed by its usage in studies of cardiovascular disease and how it pertains to viral and parasitic infections of the heart. PMID- 16368558 TI - Apoptosis Gene Information System--AGIS. AB - Genes implicated in apoptosis have great relevance to biology, medicine and oncology. Here, we describe a unique resource, Apoptosis Gene Information System (AGIS) that provides data for over 2400 genes involved directly or indirectly, in apoptotic pathways of more than 350 different organisms. The organization of this information system is based on the principle of one-gene, one record. AGIS will be updated on a six monthly basis as new information becomes available. AGIS can be accessed at: http://www.cellfate.org/AGIS/. PMID- 16368559 TI - Biomolecular markers of breast cancer. AB - Here, the structure, function, biological and pathological significance and clinical utility of the principal biomolecular markers of breast cancer is reviewed. Each marker was scored for clinical utility using a recently developed tumor marker utility grading system (TMUGS). Among the tissue markers, ERs and PRs are important prognostic/predictive factors and the only tissue markers routinely determined. ER cross-talks with other growth factors while co regulatory factors enhance (co-activators) or decrease (co-repressors) its transcriptional activity. C-erbB-2 and Ki67/MIB-1 select for adjuvant chemotherapy a subgroup of lymph-node negative patients at a high risk of relapse. Monoclonal antibodies (trastuzumab, gefitinib, erlotinib and bevacizumab) targeting tissue markers and involved in tumor growth and metastasization (EGFR, C-erbB-2, VEGF) have been developed; they showed therapeutical single agent activity as well as potent synergy with chemotherapy agents in metastatic cancer. Among circulating markers, some are potentially useful in the early detection and monitoring of metastatic disease; nevertheless, none is routinely recommended. To suspect distant metastases, CEA-TPA-CA15.3 panel attained accuracy of about 90%. ECD HER2-neu, p53 and nucleophosmin antibodies seem suitable candidates for different associations. Preliminary observations suggest that an early detection with tumor markers and successive treatment of relapses significantly prolongs disease-free and overall survival in selected patients. In conclusion, biomolecular markers are improving understanding of biology and management of breast cancer. PMID- 16368560 TI - Molecular cytogenetic characterization of a novel cell line established from a superficial spreading melanoma. AB - We report the complex cytogenetic analysis of a novel melanoma cell line (M35/01) established from a vertical growth phase of a superficial spreading melanoma. Similarly to its parental tumor, this cell line metastasizes to the liver. Using combined molecular cytogenetic techniques, we could identify a reservoir of chromosomal alterations in M35/01. In addition, we had sufficient amount of DNA from both the original primary tumor and the cell line which allowed for the comparison of their genetic patterns by chromosomal CGH. Several common alterations were found indicating the same clonal origin. These alterations included gains of 6p, 7q, 15q and deletions of 9, 10, 16q and 17p. Chromosomal losses present only in the cell line were detected on chromosome 4, 16p, 18 and gains on 20p12-qter. Array CGH analysis of the M35/01 cell line provided similar results with a much higher resolution, representing relatively high level gains on 7q31.2-q31.31, 15q25, 20q, and losses on 4q28, 9p21-p24, 9q21-q22, 10q25, 16q13-q23, 17p12-13 and 18q12-23. Using SKY-FISH, several structural alterations could be detected which were not recognized by conventional cytogenetics. Except for chromosome 18, none of the centromeres showed normal distribution by FISH. Our analysis shows that a high number of chromosomal alterations, which are known to be nonrandomly associated with melanoma progression, can be found by the combined use of different molecular genetic techniques. This new melanoma cell line would be an excellent model for investigating the mechanism of organ specific-metastatic events of malignant melanoma. PMID- 16368561 TI - The genetic basis for bronchopulmonary dysplasia. AB - While the 'original' bronchopulmonary dysplasia (BPD) was attributed to the iatrogenic effects of oxygen and barotrauma on the preterm lung, analyses of the 'new' BPD suggests that these environmental effects may contribute to arrested pulmonary development, and that there may also be genetic foundations for the susceptibility to BPD. Twinning, family and population studies implicate heritable factors in the evolution of BPD. The candidate genes examined for their potential role in BPD include surfactant apoprotein and inflammatory genes. With the identification and mapping of single nucleotide polymorphisms (SNPs), an explosion of testing for these genetic components that may contribute to a number of complex, multigenic disease conditions-including BPD-have been initiated. Sophisticated multiplex analyses are now available to link candidate SNPs to conditions such as BPD. However, there continues to be wide variation in the expression of BPD throughout neonatal units. Differentiating the effects caused by environmental and environmental-genetic interactions from isolated genetic etiologies is still problematic and will require carefully designed genetic analyses of preterm infant groups and their families. PMID- 16368562 TI - Expression of tissue inhibitor of metalloproteinase during early stages of bone graft healing. AB - The aim of this study was to investigate the temporal expression of TIMP-1 within endochondral and intramembranous bone grafts during the early stages of healing in thirty six adult New Zealand white rabbits. Total RNA was isolated from bone grafts extracted on days 0-11 and day 14 post-grafting, for RT-PCR analysis. In situ hybridization was carried out on days 1-9 and day 14. Results showed TIMP-1 expression coincides with osteogenesis, which indicates a role for TIMP in preserving the newly formed bone during the initial stages of graft healing. Bone grafts play an important role in influencing the healing process mediated by the host tissues. The temporal expression of TIMP-1 differs in endochondral and intramembranous bone grafts. The earlier expression of TIMP-1 by endochondral bone grafts, could be the reason for the delayed vascularization while the expression of TIMP-1 by the intramembranous bone grafts, at a later stage could allow for earlier vascularization. PMID- 16368563 TI - Echinomycin and cobalt-phenanthroline as redox indicators of DNA hybridization at gold electrodes. AB - A bis-intercalator echinomycin (ECHI) and a simple intercalator [Co(phen)3]3+ were used as a novel electrochemical redox indicators to detect DNA hybridization at gold electrodes (AuE). In order to minimize the nonspecific adsorption of oligonucleotides (ODN), the thiol-derivatized oligonucleotides were immobilized onto AuE in the first step, and the exposition of AuE to 6-mercapto-1-hexanol (MCH) followed in the second step of this procedure. In this arrangement good reproducibility and discrimination between single-stranded (ss) probe and double stranded (ds) hybrid DNA were obtained. While both redox indicators showed a good ability to discriminate between the ss probe and ds hybrid DNA, the signals of ECHI were by an order of maginitude higher than those of [Co(phen)3]3+ in a good agreement with stronger DNA binding by the bis-intercalator as compared to the simple intercalator. In addition, DNA single-base mismatch (DNA point mutation) was easily detected by means of ECHI. PMID- 16368564 TI - CRH-BP: the regulation and function of a phylogenetically conserved binding protein. AB - Corticotropin Releasing Hormone-Binding Protein (CRH-BP), a 37 kDa secreted glycoprotein, binds both CRH and urocortin with high affinity and is structurally unrelated to the CRH receptors. CRH-BP orthologues have been identified in multiple invertebrate and vertebrate species. It is strongly conserved throughout evolution, suggesting the maintenance of a structural conformation necessary for biological activity. CRH-BP is an important modulator of CRH activity; it inhibits CRH-induced ACTH secretion from pituitary corticotropes and may exert similar actions at central sites of CRH release. While the function of CRH-BP is thought to be primarily inhibitory, recent studies indicate that novel functional roles may exist in both the brain and pituitary. Regulation of CRH-BP expression by stress and metabolic factors are consistent with in vivo models of altered CRH BP expression. Positive regulation of pituitary CRH-BP by reproductive hormones suggests that additional interactions between the stress and reproductive axes may exist. While recent research has focused on the evolutionary conservation, expanded sites of expression, regulation and in vivo function of CRH-BP, a more complete understanding of the central and peripheral functions of CRH-BP and its mechanisms of action will help elucidate its potential role in the etiology or treatment of disorders of CRH dysregulation. PMID- 16368565 TI - Dynamic changes in NuMA and microtubules in monkey-rabbit nuclear transfer embryos. AB - Previous reports have indicated that failure in cloning monkey is attributed to the removal of nuclear mitotic apparatus (NuMA) during enucleation and subsequent abnormal organization of mitotic apparatus. This study investigated the transformation and assembly of tubulin and NuMA protein during the first cell cycle of cloned monkey embryos reconstructed by using enucleated rabbit oocytes as recipients. After the oocyte fused with a fibroblast, extensive microtubule organization was observed around the introduced nucleus in most reconstructed embryos, suggesting the introduction of a somatic cell centrosome. A high proportion of fibroblast nuclei transferred into non-activated oocytes underwent premature chromosome condensation (PCC), transient spindle organization and chromosomes separation, followed by the formation of two pronucleus-like structures. In contrast, fibroblast nuclei in pre-activated ooplasm rarely underwent PCC, but formed a swollen pronucleus-like structure. Normal spindles were observed in about one third of the cloned embryos reconstructed by both methods. After transferring monkey fibroblasts into NuMA-removed enucleated rabbit oocytes, NuMA was localized in pseudo-pronuclei and gradually moved to mitotic spindle poles at the first mitotic spindle poles. NuMA antibody microinjection resulted in spindle disorganization and chromosome misalignment, but did not significantly affect early cleavage. Our findings indicate that: 1. NuMA in donor monkey fibroblast may contribute to form a normal spindle in enucleated rabbit oocyte; 2. when non-activated cytoplasts and pre-activated cytoplasts are used as recipients, the donor nuclei undergo different morphological changes, but yield similar early embryo development; 3. although abnormal spindle organization and chromosome alignment may cause low efficiency of animal cloning, these abnormalities do not significantly affect early cleavage. PMID- 16368567 TI - A novel bacterium Saprospira sp. strain PdY3 forms bundles and lyses cyanobacteria. AB - A helical filamentous cyanobactericidal bacterium was isolated from Dianchi Lake, a eutrophic freshwater lake in Kunming City of the Yunnan Province in China using a special solid medium. This species was designated strain PdY3. This bacterium was identified as a novel Saprospira sp. on the basis of its morphological characteristics and 16S rDNA sequence. Strain PdY3 showed apparent group behavior on the solid medium, forming orderly, bundle-like group structures. These bundles moved as groups. Individuals in a bundle responded to the bundle as a whole. PdY3 also showed group behavior and formed a three-dimensional reticular structure when co-cultured with Anabaena in liquid media. This helical bacterium lysed cyanobacteria through direct contact and its group behavior greatly accelerated the cyanobactericidal process. Our experiments showed that PdY3 caused lysis of 64% of Anabaena cells within 1 day and that its cyanobactericidal range was broad. These results underscore potential application of Saprospira on the control of blooms of cyanobacteria. PdY3 group behavior might allow a more efficient capture of bacterial prey. PMID- 16368566 TI - Wnt/Frizzled signaling in hepatocellular carcinoma. AB - The Wnt/Frizzled (FZD) signaling cascade is important for cell fate determination during embryonic development as well as maintaining tissue homeostasis in the adult. In addition to these physiologic roles, studies have shown that deregulation of Wnt/FZD signaling occurs during carcinogenesis. As an example, over 90% of the colorectal cancers have mutations in adenomatous polyposis coli (APC) or beta-catenin genes. In addition, hepatocellular carcinoma (HCC) is another tumor with frequent aberrant activation of beta-catenin signaling. Nuclear and/or cellular beta-catenin accumulation, a hallmark of the activated canonical Wnt/FZD signaling, has been observed in 33-67% of tumors. However, mutations of APC and/or beta-catenin genes are found only in about 20-30% of HCCs, suggesting that the predominant mechanism(s) activating Wnt/FZD signaling pathway may be different from that found in colorectal cancers. There is accumulating evidence to suggest that regulatory mechanisms other than mutations involving beta-catenin or proteins in its destruction complex, many of which involve upstream components of the Wnt/FZD cascade, are important in HCC. Furthermore, information on the target genes of Wnt/FZD signaling and their roles in hepatocarcinogenesis is limited despite the recent discovery of several candidate genes. This review focuses on the alterations of Wnt/FZD signaling pathways and their relationship to the pathogenesis of HCC. A better understanding of the precise mechanisms of altered Wnt/FZD signaling may provide new molecular targets for therapy of HCC. PMID- 16368568 TI - Analysis of common gene expression patterns in four human tumor cell lines exposed to camptothecin using cDNA microarray: identification of topoisomerase mediated DNA damage response pathways. AB - Camptothecin (CPT) is a potent inhibitor of DNA topoisomerase I with a wide spectrum of anti-tumor activity. Relatively little information is available regarding the relation of known topoisomerase-mediated DNA damage with other intracellular pathways. To gain an insight into the intracellular molecular mechanisms of Topoisomerase I inhibitor camptothecin-mediated DNA damage leading to cell death, we used a high-density cDNA microarray to assess sensitive early gene expression profiles in SGC7901 (gastric cancer), Hela (cervical adenocarcinoma), K562 (chronic myelogenous leukemia) and HL60 (promyelocytic leukemia) tumor cells stimulated with camptothecin for 1 h at the concentrations of GI50 (50 % growth inhibition after 24 h of treatment). Analysis of the differentially expressed genes obtained 29 response genes common to all four cell lines. Moreover, these cell lines also shared the direction of regulation. Most of these common response genes were functionally related to cell proliferation or apoptosis, and some of them were involved in ATM (ataxia-telangiectasia mutated) and ATR (ATM-and Rad3 related) checkpoint pathways, JNK (c-Jun N-terminal kinase) pathway, the survival phosphatidylinositol (PI) 3 kinase-Akt-dependent pathway, mitochondrial cell death pathway, endoplasmic reticulum (ER)-related cell death pathway, and to ubiquitin/proteasome dependent protein degradation pathway. The data provides evidence for a linkage between topoisomerase-mediated DNA damage and intracellular signaling events, which may facilitate our understanding of the camptothecin mediated molecular mechanisms of action. PMID- 16368569 TI - Laboratory analysis of T-cell immunity. AB - Immune-based strategies for treating and preventing cancer are increasingly being tested and include cancer vaccination, adoptive T cell therapy, and cytokine therapy. An important component of testing and development of immune-based strategies is monitoring the immunologic response. The ability to monitor T cell immunity has been suboptimal. The measurement of tumor-specific immunity will aid in defining which strategies should be moved forward in clinical trials and which should be eliminated or evaluated further in the preclinical realm. Immunologic monitoring is necessary for determining if an approach has immunologic efficacy and ultimately whether immunologic responses correlate with a clinical response. This article discusses several important elements of measuring T cell immunity such as validation principles, laboratory issues, current approaches, and new paradigms and concepts for future testing. Informative immunologic monitoring of T cells will be one of the driving forces in advancing the field of tumor immunology. PMID- 16368570 TI - The Nuclear Mitotic Apparatus (NuMA) protein is contributed by the donor cell nucleus in cloned porcine embryos. AB - The Nuclear Mitotic Apparatus (NuMA) protein is a multifunctional protein that is localized to the nucleus in interphase and to the poles of the mitotic apparatus during mitosis. In unfertilized porcine oocytes, NuMA is localized to the meiotic spindle. NuMA is removed along with the meiotic spindle during the enucleation process before reconstructing the egg by introducing the donor cell nucleus to produce cloned embryos. Questions have been raised regarding the source for NuMA in cloned embryos, as the enucleated oocyte does not contain detectable NuMA in the cytoplasm. To determine the source of NuMA in porcine nuclear transfer (NT) embryos, we conducted an immunofluorescence microscopy study with antibodies against NuMA to investigate the appearance and distribution of NuMA before and after reconstructing NT embryos with porcine skin fibroblasts. We used donor cells from a confluent culture with all cells in interphase. For comparative studies, we also determined the immunofluorescence pattern of NuMA, gamma tubulin, and alpha-tubulin in porcine fibroblasts, parthenogenetic embryos and in vitro fertilized (IVF) embryos. Results show that NuMA was localized in nuclei of 33.5% (163/456) of the serum-deprived fibroblasts used as donor cells. No NuMA staining was detected in enucleated pig oocytes. Immediately after nuclear transfer, NuMA staining was absent in all donor cell fibroblast nuclei (0 h) but staining was detected by 6 h within the reconstructed eggs, at which time the transferred somatic cell nucleus swelled in most cells (19/27) and became a pronucleus-like structure. NuMA was localized exclusively within the pronucleus like structures (15/27). At 25 h, NuMA was detected inside the nucleus (16/25) either in one-cell or in 2-cell stage embryos. Interestingly, in parthenogenetic embryos, NuMA staining was not detected in all 42 eggs examined at 1 h, and evident NuMA staining was only detected inside a few (4/51 at 6 h; 6/48 at 25 h) of the nuclei. In IVF embryos, NuMA was detected within the nucleus at 6 h (5/20) and 25 h (13/16). These results show that the donor cell nucleus contains NuMA that is contributed to the reconstructed embryo and possibly activated by mechanisms in the oocyte's cytoplast. PMID- 16368571 TI - Mechanisms of eukaryotic DNA double strand break repair. AB - For all cells, a DNA double strand break (DSB) is a dangerous lesion that can have profound consequences for the genome. If a DSB is encountered during mitosis, chromosomal separation may be adversely affected. Alternatively, during S phase a DSB may cause replication fork stalling or collapse. Improperly repaired DSBs can result in chromosomal rearrangements, senescence or activation of apoptotic pathways. Cells have developed sophisticated recombination pathways to metabolize and repair DSBs quickly as well as the capacity to differentiate physiologically occurring breaks from life threatening lesions. The two major pathways of recombination repair are known as non-homologous end-joining (NHEJ) and homologous recombination (HR). In this review, we will discuss the detection, response, and repair of DSBs in eukaryotes. PMID- 16368572 TI - The role of ADP receptors in platelet function. AB - Adenine di-Phosphate (ADP) is an important physiological agonist that plays a vital role in normal hemostasis and thrombosis. The importance of ADP in normal hemostasis is clearly demonstrated in patients suffering from storage pool disease who show excessive bleeding tendencies. It is well established that ADP activates platelets through 3 purinergic receptors, namely P2Y1, P2Y12 and P2X1. The P2Y1 receptor is a Galpha q coupled G-protein receptor that is important for platelet shape change, aggregation, thromboxane A2 generation, procoagulant activity, adhesion to immobilized fibrinogen and thrombus formation under shear conditions. The availability of P2Y1 antagonists and knockout mice have aided in demonstrating the multiple functions of this receptor in platelet function and normal hemostasis. The second ADP receptor, the P2Y12 signals through a Galpha i coupled G-protein receptor and has been shown to be important for platelet functions very similar to the P2Y1 receptor. In addition, the P2Y12 receptor is also important for potentiation of platelet activation mediated by other physiological agonists including collagen, von Willebrand and thromboxane A2. Advances in understanding the importance of the P2Y12 receptor has resulted in the development of drugs like clopidogrel and ticlopidine that is being successfully used clinically in the treatment of thrombotic disorders. The understanding of the function of this receptor has been possible due to the availability of multiple P2Y12 antagonists and the development of the P2Y12 null mice. The third and the final P2X1 receptor is an ion channel that upon activation causes an influx of calcium. Even though the activation of this receptor by itself doesn't lead to aggregation, it causes shape change and aids in the activation process of other agonists. Studies have also shown that it is important for thrombus formation under shear conditions in small arteries. Signaling by this receptor leads to significant ERK activation, which has been shown, to be important for collagen mediated platelet activation. The importance of ADP in hemostasis and thrombosis greatly underscores the significance of understanding the function of these receptors that would enable development of potent and safe anti-thrombotic drugs. PMID- 16368573 TI - Role of ERM (ezrin-radixin-moesin) proteins in T lymphocyte polarization, immune synapse formation and in T cell receptor-mediated signaling. AB - Following antigen recognition, T lymphocytes undergo strong actin cytoskeletal rearrangements. These play a crucial role in the molecular reorganization at the contact site between the T lymphocyte and the antigen presenting cell, termed the immunological synapse. Moreover, they are necessary for T cell activation that leads to cytokine secretion, T cell proliferation and effector function. Little is known on how membrane and signaling molecules interact with the actin cytoskeleton during these processes. Here we review the function of the ERM family of membrane-microfilament linkers, making emphasis on the role of these proteins in T lymphocyte physiology. We discuss how ERM proteins are involved in membrane reorganization during T lymphocyte polarization and immune synapse formation, and how these proteins may contribute to T cell receptor-mediated intracellular signaling that leads to T cell activation. PMID- 16368574 TI - Micro-RNA-like effects of complete intronic sequences. AB - MicroRNAs (miRNAs) have been suggested as suppressors of numerous target genes in human cells. In this report, we present gene chip array data indicating that in the absence of miRNA sequences, complete human introns are similarly capable of coordinating expression of large numbers of gene products at spatially diverse sites within the genome. The expression of selected intronic sequences (6a, 14b and 23) derived from the cystic fibrosis transmembrane conductance regulator (CFTR) gene caused extensive and specific transcriptional changes in epithelial cells (HeLa) that do not normally express this gene product. Each intron initiated a distinctive pattern of gene transcription. Affected genes such as FOXF1, sucrase-isomaltase, collagen, interferon, complement and thrombospondin 1 have previously been linked to CFTR function or are known to contribute to the related processes of epithelial differentiation and repair. A possible regulatory function of this nature has not been demonstrated previously for non-coding sequences within eukaryotic DNA. The results are consistent with the observation that splicesomal introns are found only in eukaryotic organisms and that the number of introns increases with phylogenetic complexity. PMID- 16368575 TI - Chimeric reality and myth. PMID- 16368576 TI - First trimester maternal blood rheology and pregnancy induced hypertension. AB - This study evaluates the relationship between the first trimester assessment of maternal rheology and the subsequent development of pregnancy induced hypertension. This is a prospective observational study based in the Glasgow Royal Maternity Hospital, Scotland. From an original population of 744 consecutive antenatal attendees a total of 579 women were booked at less than 14 weeks' gestation. The main study group is a further subset comprising 251 primigravid women booking with a singleton pregnancy without essential hypertension. Previously published data from a group of non-pregnant women of similar age drawn from the same local community was used for external comparison. Blood samples were collected at the booking visit, from which fibrinogen, red cell aggregation, haematocrit and plasma, whole blood, relative and corrected viscosities were recorded. Information was obtained from the case notes in retrospect starting approximately 1 year after the first patients had first been recruited into the trial. The overall outcome of the pregnancies was noted with particular reference to pregnancy induced hypertension (PIH), birth weight, antepartum haemorrhage, pre-term labour, perinatal death, condition at delivery and neonatal complication. Our results show PIH is associated with a significantly raised mean blood viscosity and fibrinogen at time of booking. All significance disappears after adjustment for smoking, diastolic blood pressure and age. Viscosity is, however, only marginally non-significant (p = 0.07). In conclusion, blood rheology, in particular blood viscosity and fibrinogen, may play a predictive role in the development of pregnancy-induced hypertension. When combined with measurement of smoking and diastolic blood pressure at booking, these measurements could be used to calculate a risk score for the development of PIH, allowing targeting of antenatal care. Further data is required. PMID- 16368577 TI - Bacterial vaginosis diagnosed at the first antenatal visit better predicts preterm labour than diagnosis later in pregnancy. AB - This study was conducted as part of a double-blind randomised placebo-controlled trial, the aim of which was to determine whether vitamin C could reduce the recurrence risk of pre-term labour. In this study, women with a history of pre term labour in a preceding pregnancy were randomised to receive either 250 mg vitamin C or a matching placebo twice daily until 34 weeks' gestation. They attended a dedicated pre-term labour clinic every 2 weeks. All women were screened for bacterial vaginosis (BV) at each visit. It was first determined that vitamin C did not have any effect on the presence of BV. Women who were diagnosed with BV before 20 weeks' gestation were at higher risk of delivering pre-term than those who developed BV after 20 weeks. PMID- 16368578 TI - Audit of intrapartum group B streptococcus prophylaxis. AB - Group B streptococcus is the most frequent cause of severe early onset neonatal infection. For over 3 years our department has employed a risk-factor-based protocol of intra-partum antibiotic prophylaxis. Prophylaxis has been considered adequate if women receive antibiotics more than 4 hours before delivery. This study aims (1) to establish the adequacy of intrapartum prophylaxis in routine clinical practice and (2) to determine whether changing the time interval (from commencement of prophylaxis to delivery) for adequacy from 4 to 2 hours (based on RCOG guidelines) improves the percentage of women receiving adequate prophylaxis. This was an audit against established protocol. Case notes of consecutive deliveries over a 4-week period were reviewed retrospectively. A total of 396 deliveries occurred during the study period. Some 44 women (11%) were eligible for antibiotic prophylaxis. Adequate prophylaxis was achieved in 21 cases (49%). If the time for prophylaxis to be considered adequate was reduced from 4 to 2 h, the number of women receiving adequate prophylaxis increased to 33 (76%). In conclusion, despite the protocol being well established, only half of the eligible women receive adequate intra-partum prophylaxis. Reducing the time interval from 4 to 2 h appreciably increases the number of women receiving adequate prophylaxis, which reduces the necessity of providing neonatal prophylaxis. PMID- 16368579 TI - A comparison of the effectiveness of single-dose vs multi-dose antenatal corticosteroids in pre-term neonates. AB - The purpose of this study was to compare the outcome between exposure to single vs multiple courses of antenatal corticosteroids (AC) in pre-term births. The study involved 704 neonates delivered between 24-32 weeks. Maternal and perinatal outcome were compared between 294 (42%) neonates who received no AC; 257 (36%) who received single AC and 153 (22%) who received multiple AC. Any AC compared with no AC showed decreased composite neonatal morbidity (51% vs 62%, p = 0.004, odds ratio (OR) = 0.6), neonatal death (52% vs 62%, p = 0.004, OR = 0.6) and hyaline membrane disease (45% vs 57%, p = 0.002, OR = 0.6) with increased chorioamnionitis (17% vs 11%, p = 0.037, OR = 1.6) and pulmonary oedema (12% vs 1%, p = 0.0001, OR = 13). Multiple AC compared with single AC was associated with increased positive maternal cultures (44% vs 31%, p < or =0.005, OR = 1.8), small for gestational age infants (35% vs 21%, p = 0.001, OR = 2) and intraventricular haemorrhage (45% vs 34%, p <0.05, OR = 1.6). Multiple corticosteroid courses when compared with single corticosteroid course did not further reduce composite neonatal morbidity but increased the risk of positive maternal cultures and neonatal small for gestational age and intraventricular haemorrhage. PMID- 16368580 TI - The bacteriology of pre-labour rupture of membranes in a Nigerian teaching hospital. AB - A prospective case control study that was conducted at the University of Ilorin Teaching Hospital, Ilorin, Nigeria, between 1st January and 31st December 2002. The purpose of this study was to determine the association and the pattern of bacteria/microorganisms in the aetiology of pre-labour premature rupture of membrane (PROM) in this centre. A total of 108 cases of PROM and 98 control cases that presented between 37 completed weeks' and 40 weeks' gestation were analysed. Pathogens were isolated in 48 patients, giving a recovery rate of 44.4%. The common pathogens include Gardnerella vaginalis (29.1%), Candida (23.0%) and Staphylococcus aureus (18.7%). Others were Streps. Pyogenes (16.6%), coagulase negative staphylococcus (CONS) (6.3%) and Klebsiella (6.3%). Only Candida and S. aureus were isolated in the controls. Ofloxacin and azithromycin were 100% active against all the isolated pathogens, while ampicillin was the least active. G. vaginalis was the most sensitive among the isolates while CONS and Klebsiella were the least sensitive. It is evident in this study that some pathogens were associated with PROM and that G. vaginalis was the most common organism and azithromycin was the only antibiotic with 100% sensitivity. We suggest that metronidazole should be added to azithromycin to cover for anaerobes in cases of PROM, where facilities for screening for anaerobes are not available. PMID- 16368581 TI - A retrospective review of magnetic resonance imaging in assessing the extent of myometrial infiltration for patients with endometrial carcinoma. AB - The surgical treatment of endometrial malignancy is based on pre-operative assessment of differentiation, myometrial and lymph node infiltration. Studies have reported the use of magnetic resonance imaging (MRI) with conflicting results. This study aims to review the pre-operative accuracy of MRI against the histology in endometrial cancer within a district hospital. A total of 112 women were identified from January 1997 to December 2004. The post-operative FIGO staging showed 76.2% Stage I, 7.6% Stage II, 14.3% Stage III and 1.7% Stage IV. A total of 57% (n = 60) had a pre-operative MRI. The myometrial infiltration was difficult to interpret in 31 MRIs. The time between hysteroscopy and MRI scan in those cases was shorter (Mean 3 weeks, SD +/- 1.5) than in reports with a clear interpretation (4.4 weeks, SD +/- 1.6). Sensitivity for MRI to detect deep myometrial infiltration was 35.7% (5/14) and specificity 50% (23/46). Accuracy was 46.6% (28/60). The presented data suggests a poor predictability of myometrial invasion by MRI but the numbers are small and MRI is a rapidly evolving modality. The conclusions must be treated with caution and cannot easily be applied to large gynaecological cancer units in general. PMID- 16368582 TI - Successful laparoscopic management of ectopic pregnancy in a district general hospital. AB - A retrospective analysis of the management of ectopic pregnancy was undertaken at Billinge Hospital, Wigan from June 1999 to June 2002. A total of 114 cases of ectopic pregnancy were identified. Diagnosis was usually confirmed by laparoscopy (89.4%) and 102 cases (82.4%) were managed by laparoscopic salpingectomy. A total of 7.08% of patients needed a laparotomy after the initial laparoscopy and two (1.75%) had laparotomy performed as the primary approach. Medical treatment was given to eight women (7.0%). All the consultants were competent in performing laparoscopic surgery and 71.3% of cases were performed laparoscopically by a consultant. The diagnostic accuracy was high using a combination of urine pregnancy tests, serum beta-hCG and transvaginal scan. PMID- 16368583 TI - Gender and sufferings. AB - The World Health Organization describes violence against women, as the most pervasive yet least recognised human rights abuse which cuts across geographical, racial, social and economic boundaries in the world. In the present study of the 2000 women interviewed, nearly 50% reported having been physically assaulted. Age, education or socioeconomic status did not change anything but more women from nuclear families were assaulted (52%) compared with other group. Some 68% of women were assaulted by husbands and 7% by multiple perpetrators. The reasons for assault varied. Nearly 60% of those assaulted had informed someone and in around 65% of cases someone had tried to help, but in only 50% of those where help was offered, was the situation improved. The complex problem of gender violence needs to be remembered while providing healthcare because it is common. Women need to be aware of their rights and possible help. We need to progress towards a civilised society with gender equality and no violence. PMID- 16368584 TI - The abortion paradox in Uganda: fertility regulator or cause of maternal mortality. AB - Both fertility and maternal mortality indices are high among Ugandan mothers. The expected benefits in fertility and maternal mortality reduction from a rising contraceptive uptake in the country (from 5% in 1991 to 23% by the year 2000) have not been forthcoming because the increase in contraceptive prevalence rate (CPR) was below the critical level required to cause any meaningful change in overall fertility and maternal mortality. The strong desire among couples to limit family size coupled with the lack of access to modern methods of contraception by many women, especially in the rural areas of the country, have contributed to the increasing use of abortion as a means of averting unplanned or mistimed motherhood. In contrast to the expected results of a typical fertility regulator, however, abortion seems to up-regulate instead of down-regulate the occurrence of maternal mortality. This paradoxical relationship is explained mainly by the illegality of the procedure, which converts it to a clandestine activity performed by poorly trained individuals operating, in many instances, in septic settings. A practical solution is to make modern and effective methods of contraception widely available, especially among rural-dwellers. Through this and coupled with training of personnel, as well as demystification of abortion by dismantling the stigma of "illegality" associated with it, down-regulation of fertility and maternal mortality can both be achieved in a country like Uganda where population explosion is further complicated by a high incidence of maternal demise. PMID- 16368585 TI - A prospective, randomised, single-blinded, controlled trial comparing two topical anaesthetic modalities for the application of a tenaculum to the cervix. AB - The aim of this study was to compare the efficacy of equivalent doses of lignocaine spray vs lignocaine jelly in reducing pain during the application of a tenaculum to the cervix. A total of 58 women undergoing hysterosalpingography were prospectively randomised to receive either two doses of 10% lignocaine spray or 1 ml of 2% lignocaine jelly (both doses equivalent to 20 mg of lignocaine base) topically onto the cervix before tenaculum attachment. There was no difference in pain scores (measured by visual analogue scale and 4-point verbal descriptor scale) between lignocaine spray and lignocaine jelly during the attachment of the tenaculum to the cervix. In conclusion, there was no difference in pain during tenaculum attachment to the cervix following topical application of equivalent doses of either lignocaine jelly or spray. PMID- 16368586 TI - Prevalence and correlates of depression in treatment-seeking women with vulvodynia. AB - This study aimed to examine rates and correlates of depression in a treatment seeking sample of women with vulvodynia. A total of 53 women were independently diagnosed with vulvodynia and assessed with state-of-the-art measures of major depressive disorder (MDD) and depressive symptom severity as well as psychometrically established measures of pain severity, general functioning, sexual function and quality of life. Current and lifetime prevalence rates for MDD were 17% (n = 9) and 45% (n = 24), respectively. Women with current MDD reported significantly greater pain severity, and worse functioning and quality of life than women without current MDD. Among those with lifetime MDD, the majority (62.5%) reported that their first depressive episode occurred before the onset of vulvodynia. Rates of current MDD appeared to be lower than rates of MDD among other samples of treatment seeking chronic pain patients. In summary, co morbid MDD is related to greater pain severity and worse functioning among women with vulvodynia. PMID- 16368587 TI - Knowledge, attitude and practice of family planning among Igbo women of south eastern Nigeria. AB - A total of 200 Nigerian women visiting Nnamdi Azikiwe University Teaching Hospital's antenatal clinic were interviewed about their knowledge, attitude and practice of family planning. About 90% were literate. Their knowledge (80%) and approval (87%) of family planning was high, but the practice of modern family planning was low (25%) with most women involved in Billings/safe period (56%). The common methods used were Billings/safe period, condom, withdrawal and the intrauterine contraceptive device (IUCD). A total of 81.5% of the respondents are still willing to give birth while 77% agreed that their last pregnancy was planned. A total of 58.5% of respondents were educated about family planning in the antenatal clinic. The most common source of family planning information was mass media, closely followed by health workers, while the most common single reason for non-practice of family planning was rejection by the husband. We therefore conclude that despite the high education/literacy with the attendant and high knowledge and approval rate of family planning in this part of Nigeria, the practice of family planning is still low, especially due to partner objection. Policy makers should therefore increase male involvement in family planning programmes and pursue a more aggressive public awareness campaign. PMID- 16368588 TI - Changes in sexual practices and responses among ante-natal clinic attendees in a Nigerian teaching hospital. AB - A cross-sectional study was carried out among 134 antenatal clinic attendees in a Nigerian tertiary hospital to assess pregnancy-related sexual beliefs and changes in sexual frequency and response. Information was collected through a semi structured questionnaire, and analysed by SPSS. Only 15% of women believed that religious, social or cultural reasons prevented sexual intercourse in pregnancy. Frequency of sexual intercourse decreased in pregnancy in 37.4% of the respondents, remained unaltered in 46.1% and increased in 16.5%. Age, marriage duration and gestational age were not associated with change in the pattern of coital frequency in pregnancy, but education was significantly associated. Sexual responsiveness diminished in approximately half of our respondents in terms of arousal (54.5%), orgasm (48.5%), pleasure (43.7%) and satisfaction (51.4%). The changes were not associated with pregnancy duration. We concluded that sex in pregnancy is well accepted in our environment, and health workers should promote sexual health and well-being in pregnancy. PMID- 16368589 TI - Assessment of tubal factor contribution to female infertility in a low resource setting (southwest Nigeria): hysterosalpingography vs laparoscopy. PMID- 16368590 TI - Caecal volvulus secondary to malrotation presenting after caesarean section. PMID- 16368591 TI - Group A streptococcal puerperal sepsis: the return of a potentially fatal disease. PMID- 16368592 TI - First trimester superior sagittal sinus venous thrombosis and antithrombin deficiency. PMID- 16368593 TI - Renal arteriovenous malformation complicating pregnancy. PMID- 16368594 TI - Pentalogy of Cantrell: diagnosis in the first trimester. PMID- 16368595 TI - An unusual case involving a tear in the uterosacral ligament following a prolonged second stage of labour. PMID- 16368596 TI - Radial nerve injury in association with automated blood pressure recording in labour. PMID- 16368597 TI - In-utero intracranial haemorrhage probably secondary to domestic violence: case report and literature review. PMID- 16368598 TI - Geleophysic dysplasia: a patient with a severe form of the disorder. PMID- 16368599 TI - An incidental finding of persistent trophoblastic tissue. PMID- 16368600 TI - Testosterone secreting endometrioid adenocarcinoma of the ovary. PMID- 16368601 TI - A uterus didelphys bicollis, with endometrial cancer in both uteruses. PMID- 16368602 TI - Endometrial adenocarcinoma following ThermaChoice balloon endometrial ablation. PMID- 16368603 TI - Malignant melanoma of the uterine cervix. PMID- 16368604 TI - Incarcerated endometriotic ovarian cyst within paraumbilical hernia. PMID- 16368605 TI - The laparoscopic appearances of pelvic splenosis: a case of mistaken identity. PMID- 16368606 TI - Fundal leiomyoma presenting as acute on chronic uterine inversion. PMID- 16368608 TI - Exercise and the cytokine balance: a paediatric perspective. PMID- 16368609 TI - Combined effects of pre-cooling and water ingestion on thermoregulation and physical capacity during exercise in a hot environment. AB - The aim of the present study was to determine the combined effects of pre-cooling and water ingestion on thermoregulatory responses and exercise capacity at 32 degrees C and 80% relative humidity. Nine untrained males exercised for 60 min on a cycle ergometer at 60% maximal oxygen uptake (VO2max) (first exercise bout) under four separate conditions: No Water intake, Pre-cooling, Water ingestion, and a combination of pre-cooling and water ingestion (Combined). To evaluate the efficacy of these conditions on exercise capacity, the participants exercised to exhaustion at 80% VO2max (second exercise bout) following the first exercise bout. Rectal and mean skin temperatures before the first exercise bout in the Pre cooling and Combined conditions were significantly lower than in the No Water and Water conditions. At the end of the first exercise bout, rectal temperature was lower in the Combined condition (38.5 +/- 0.1 degrees C) than in the other conditions (No Water: 39.1 +/- 0.1 degrees C; Pre-cooling: 38.7 +/- 0.1 degrees C; Water: 38.8 +/- 0.1 degrees C) (P < 0.05). Heat storage was higher following pre-cooling than when there was no pre-cooling (P < 0.05). The final rectal temperature in the second exercise bout was similar between the four conditions (39.1 +/- 0.1 degrees C). However, exercise time to exhaustion was longer (P < 0.05) in the Combined condition than in the other conditions. Total sweat loss was less following pre-cooling than when there was no pre-cooling (P < 0.001). Evaporative sweat loss in the Water and Combined conditions was greater (P < 0.01) than in the No Water and Pre-cooling conditions. Our results suggest that the combination of pre-cooling and water ingestion increases exercise endurance in a hot environment through enhanced heat storage and decreased thermoregulatory and cardiovascular strain. PMID- 16368610 TI - Impact phase kinematics of instep kicking in soccer. AB - The purpose of this study was to capture the lower limb kinematics before during and after ball impact of soccer kicking by examining the influence of both sampling rate and smoothing procedures. Nine male soccer players performed maximal instep kicks and the three-dimensional leg movements were captured at 1000 Hz. Angular and linear velocities and accelerations were determined using four different processing approaches: processed using a modified version of a time-frequency filtering algorithm (WGN), smoothed by a second-order low-pass Butterworth filter at 200 Hz cut-off (BWF), re-sampled at 250 Hz without smoothing (RSR) and re-sampled at 250 Hz but filtered by the same Butterworth filter at 10 Hz cut-off (RSF). The WGN approach appeared to establish representative kinematics, whereas the other procedures failed to remove noisy oscillation from the baseline of signal (BWF), lost the peaks of rapid changes (RSR) or produced totally distorted movement patterns (RSF). The results indicate that the procedures used by some previous studies may have been insufficient to adequately capture the lower limb motion near ball impact. We propose a new time frequency filtering technique as a better way to smooth data whose frequency content varies dramatically. PMID- 16368611 TI - Effects of swing-weight on swing speed and racket power. AB - Measurements are presented of the speed at which six different rods could be swung by four male students. Three of the rods had the same mass but their swing weight (i.e. moment of inertia) differed by large factors. The other three rods had the same swing-weight but different masses. Our primary objective was to quantify the effects of mass and swing-weight on swing speed. The result has a direct bearing on whether baseball, tennis, cricket and golf participants should choose a heavy or light implement to impart maximum speed to a ball. When swinging with maximum effort, swing speed (V) was found to decrease as swing weight (Io) increased, according to the relation V = C/Ion, where C is a different constant for each participant and n = 0.27 when Io > 0.03 kg x m2. Remarkably similar results were obtained previously with softball bats (where n = 0.25) and golf clubs (where n = 0.26). Swing speed remained approximately constant as swing mass increased (when keeping swing-weight fixed). The implications for racket power are discussed. PMID- 16368612 TI - Contributions of joint rotations to racquet speed in the tennis serve. AB - The purpose of this study was to measure the contributions of the motions of body segments and joints to racquet head speed during the tennis serve. Nine experienced male players were studied using three-dimensional film analysis. Upper arm twist orientations were calculated with two alternative methods using joint centres and skin-attached markers. The results showed that skin-attached markers could not be used to calculate accurate upper arm twist orientations due to skin movement, and that the use of joint centres produced errors of more than 20 degrees in the upper arm twist orientation when the computed elbow flexion/extension angle exceeded 135 degrees in the final 0.03 s before impact. When there were large errors in the upper arm twist orientation, it was impossible to obtain accurate data for shoulder or elbow joint rotations about any axis. Considering only the contributors that could be measured within our standards of acceptable error, the approximate sequential order of main contributors to racquet speed between maximum knee flexion and impact was: shoulder external rotation, wrist extension, twist rotation of the lower trunk, twist rotation of the upper trunk relative to the lower trunk, shoulder abduction, elbow extension, ulnar deviation rotation, a second twist rotation of the upper trunk relative to the lower trunk, and wrist flexion. The elbow extension and wrist flexion contributions were especially large. Forearm pronation made a brief negative contribution. Computed contributions of shoulder internal rotation, elbow extension and forearm pronation within the final 0.03 s before impact were questionable due to the large degree of elbow extension. Near impact, the combined contribution of shoulder flexion/extension and abduction/adduction rotations to racquet speed was negligible. PMID- 16368613 TI - The energetics and benefit of an arm swing in submaximal and maximal vertical jump performance. AB - The aims of this study were to investigate the energy build-up and dissipation mechanisms associated with using an arm swing in submaximal and maximal vertical jumping and to establish the energy benefit of this arm swing. Twenty adult males were asked to perform a series of submaximal and maximal vertical jumps while using an arm swing. Force, motion and electromyographic data were recorded during each performance and used to compute a range of kinematic and kinetic variables, including ankle, knee, hip, shoulder and elbow joint powers and work done. It was found that the energy benefit of using an arm swing appears to be closely related to the maximum kinetic energy of the arms during their downswing, and increases as jump height increases. As jump height increases, energy in the arms is built up by a greater range of motion at the shoulder and greater effort of the shoulder and elbow muscles but, as jump height approaches maximum, these sources are supplemented by energy supplied by the trunk due to its earlier extension in the movement. The kinetic energy developed by the arms is used to increase their potential energy at take-off but also to store and return energy from the lower limbs and to "pull" on the rest of the body. These latter two mechanisms become more important as jump height increases with the pull being the more important of the two. We conclude that an arm swing contributes to jump performance in submaximal as well as maximal jumping but the energy generation and dissipation sources change as performance approaches maximum. PMID- 16368614 TI - Relationships between cohesion, collective efficacy and performance in professional basketball teams: an examination of mediating effects. AB - The main aim of this study was to examine mediating effects in the relationships between cohesion, collective efficacy and performance in professional basketball teams. A secondary aim was to examine the correlates of collective efficacy in a professional sport. A total of 154 French and foreign professional players completed French or English versions of questionnaires about cohesion and collective efficacy. Two composite measures of individual performance were used (pre- and post-performance). Individual-level analyses were performed. Regression analyses supported two mediating relationships with collective efficacy as a mediator of the pre-performance - Group integration-task relationship, and Group integration-task as a mediator of the pre-performance - collective efficacy relationship. Statistical analyses indicated that neither Group integration-task nor collective efficacy was a better mediator in the relationship between pre performance and the other group variables. Results also revealed positive relationships between three dimensions of cohesion (i.e. Individual attractions to the group-task, Group integration-task, Group integration-social) and collective efficacy. These findings suggest that in professional basketball teams, staff members should look after athletes who perform at a lower or below their usual level because their performances might lead them into a downward cohesion - collective efficacy spiral. Staff members should also develop a high quality of group functioning, both on and off the basketball court, given its relationship with collective efficacy. PMID- 16368615 TI - Skinfold thicknesses associated with distance running performance in highly trained runners. AB - It has been postulated that additional adiposity has a negative effect on performance in heterogeneous groups of runners. Previous studies have not tested this hypothesis in homogeneous groups of elite runners. The purpose of this study was to determine whether the sum of skinfold thicknesses and specific single skinfold sites were related to competitive running performance in homogeneous groups of male and female elite athletes. In total, 184 top-class runners (130 males and 54 females) volunteered to participate in the study. Skinfolds were measured at the following sites: biceps, triceps, subscapular, pectoral, iliac crest, abdominal, front thigh and medial calf. Runners were classified into groups in accordance with their best performance times. Correlation analysis and partial correlation coefficients that controlled for age and weight were applied to each single skinfold, the sum of six skinfolds (excluding biceps) and the extremity (sum of triceps, front thigh, medial calf) to trunk (sum of subscapular, iliac crest, abdominal) ratio and performance. Performance was rated by the scoring procedures of the International Amateur Athletics Federation. In male runners, the pectoral, iliac crest, abdominal, biceps, triceps, subscapular skinfolds and the sum of six skinfolds were not associated with performance score for any of the distances. High correlations were found between the front thigh (r = 0.78, P = 0.000) and medial calf (r = 0.55, P = 0.018) skinfolds and 1500 m run time, and between the front thigh (r = 0.59, P = 0.014) and medial calf (r = 0.57, P = 0.017) skinfolds and 10,000 m run time. In female runners, the front thigh and medial calf skinfolds were highly correlated with 400 m run time (r = 0.71, P = 0.022 and r = 0.81, P = 0.005, respectively). The results of this study indicate that skinfold thicknesses in the lower limb are positively associated with running time over several distances, and may be a useful predictor of athletic performance. PMID- 16368616 TI - Visual attention in orienteers at different levels of experience. AB - This study explored how differences in orienteering experience are related to differences in how visual attention is allocated to the map, the environment and to travel. Twenty more experienced and 20 less experienced individuals orienteered while wearing a head-mounted video camera with microphone. The participants verbalized what they were attending to (map, environment or travel) at any given time. Each recorded film was coded at each point in time in terms of what the participant was attending to and whether the participant was moving or stationary. More experienced orienteers attended to the map markedly more while moving and spent less time stationary than less experienced orienteers. The participants' performance was significantly related to the ability to attend to the map while moving. The strategic control of attention is proposed to explain this ability. It is proposed that attentional training might enhance performance in sports characterized by multiple and dynamically varying elements. PMID- 16368617 TI - Benefits of an external focus of attention: common coding or conscious processing? AB - We conducted two experiments to assess the effect attentional focus has on learning a complex motor skill and subsequent performance under secondary task loading. Participants in Experiment 1 learnt a golf putting task (300 practice trials) with a single instruction to either focus on their hands (internal focus) or the movement of the putter (external focus). No group differences were evident during learning or retention. Differences between the groups were only apparent under secondary task load; the external group's performance remained robust, while the internal group suffered a drop in performance. Verbal protocols demonstrated that the internal group accumulated significantly more internal knowledge and more task-relevant knowledge in general than the external group. Experiment 2 was designed to establish whether greater internal focus knowledge or greater explicit rule build up in general was responsible for performance breakdown. Two groups were presented with a set of six internal or external rules. Again, no performance differences were found during learning or retention. During the secondary task, both groups experienced performance deterioration. It was concluded that accumulation of explicit rules to guide performance was responsible for the internal group's breakdown in performance under secondary task loading and may be responsible for some of the performance differences reported previously. PMID- 16368618 TI - Self-belief does make a difference: a reciprocal effects model of the causal ordering of physical self-concept and gymnastics performance. AB - A large body of research in support of the reciprocal effects model of causal ordering demonstrates that prior academic self-concept predicts subsequent academic achievement beyond what can be explained in terms of prior achievement. Here we evaluate the generalizability of this support for the reciprocal effects model to a physical activity context in which achievement is reflected in gymnastics skills on a standardized gymnastics performance test evaluated by expert judges. Based on the responses of 376 adolescents collected at the start (T1) and end (T2) of a gymnastics training programme, there is support for a reciprocal effects model in which there are significant paths leading from both T1 gymnastics self-concept to T2 gymnastics skills and from T1 gymnastics skills to T2 self-concept. Although there were gender and age effects (girls and older participants had better gymnastics skills, boys had higher self-concepts), multiple group structural equation models indicated that support for the reciprocal effects model generalized over responses by boys and girls. In summary, self-concept and performance are both determinants and consequences of each other. PMID- 16368619 TI - Plagiarism. PMID- 16368620 TI - Leg muscle recruitment in highly trained cyclists. AB - In this study, we examined patterns of leg muscle recruitment and co-activation, and the relationship between muscle recruitment and cadence, in highly trained cyclists. Electromyographic (EMG) activity of the tibialis anterior, tibialis posterior, peroneus longus, gastrocnemius lateralis and soleus was recorded using intramuscular electrodes, at individual preferred cadence, 57.5, 77.5 and 92.5 rev . min(-1). The influence of electrode type and location on recorded EMG was also investigated using surface and dual intramuscular recordings. Muscle recruitment patterns varied from those previously reported, but there was little variation in muscle recruitment between these highly trained cyclists. The tibialis posterior, peroneus longus and soleus were recruited in a single, short burst of activity during the downstroke. The tibialis anterior and gastrocnemius lateralis were recruited in a biphasic and alternating manner. Contrary to existing hypotheses, our results indicate little co-activation between the tibialis posterior and peroneus longus. Peak EMG amplitude increased linearly with cadence and did not decrease at individual preferred cadence. There was little variation in patterns of muscle recruitment or co-activation with changes in cadence. Intramuscular electrode location had little influence on recorded EMG. There were significant differences between surface and intramuscular recordings from the tibialis anterior and gastrocnemius lateralis, which may explain differences between our findings and those of previous studies. PMID- 16368621 TI - Effect of suspension systems on the physiological and psychological responses to sub-maximal biking on simulated smoothand bumpy tracks. AB - The aim of this study was to compare the physiological and psychological responses of cyclists riding on a hard tail bicycle and on a full suspension bicycle. Twenty males participated in two series of tests. A test rig held the front axle of the bicycle steady while the rear wheel rotated against a heavy roller with bumps (or no bumps) on its surface. In the first series of tests, eight participants (age 19-27 years, body mass 65-82 kg) were tested on both the full suspension and hard tail bicycles with and without bumps fitted to the roller. The second series of test repeated the bump tests with a further six participants (age 22-31 years, body mass 74-94 kg) and also involved an investigation of familiarization effects with the final six participants (age 21 30 years, body mass 64-80 kg). Heart rate, oxygen consumption (VO(2)), rating of perceived exertion (RPE) and comfort were recorded during 10 min sub-maximal tests. Combined data for the bumps tests show that the full suspension bicycle was significantly different (P < 0.001) from the hard tail bicycle on all four measures. Oxygen consumption, heart rate and RPE were lower on average by 8.7 (s = 3.6) ml . kg(-1) . min(-1), 32.1 (s = 12.1) beats . min(-1) and 2.6 (s = 2.0) units, respectively. Comfort scores were higher (better) on average by 1.9 (s = 0.8) units. For the no bumps tests, the only statistically significant difference (P = 0.008) was in VO(2), which was lower for the hard tail bicycle by 2.2 (s = 1.7) ml . kg(-1) . min(-1). The results indicate that the full suspension bicycle provides a physiological and psychological advantage over the hard tail bicycle during simulated sub-maximal exercise on bumps. PMID- 16368622 TI - The effects of ad libitum fluid ingestion on fluid balance during alpine skiing in recreational skiers. AB - The aim of this study was to assess the influence of ad libitum water ingestion, using a back-mounted hydration system (BMHS), on fluid balance during alpine skiing. Fourteen skiers skied on two different days. On one day, seven skiers ingested water during skiing via the BMHS and the other seven skiers refrained from fluid ingestion during skiing until the midday break (NW trial). On the second day, the trials were reversed. Results indicated that when skiers used the BMHS they drank significantly more water than during the NW trials (2.0 +/- 0.9 vs. 0.78 +/- 0.4 litres). However, skiers drank significantly more at the midday break during the NW trials than during the BMHS trials (0.78 +/- 0.4 vs. 0.4 +/- 0.2 litres). Percent change in plasma volume was less during the BMHS trials than during the NW trials (-0.1 +/- 5.3 vs. -4.9 +/- 5.2%), urine osmolality was maintained in the BMHS trials but rose from 295 +/- 80 to 818 +/- 168 mOsm . kg( 1) at midday during the NW trials, and body mass loss was minimized during the BMHS trials compared with the NW trials (0.4 +/- 0.4 vs. 1.1 +/- 0.2 kg). Skiers reported that they felt significantly better when they ingested water during the BMHS trials. In conclusion, a back-mounted hydration system allowed the skiers to maintain hydration status. PMID- 16368623 TI - A new device for evaluating distance and directional performance of golf putters. AB - The purpose of this study was to construct and evaluate the reliability of an apparatus for testing golf putters with respect to distance and direction deviation at different impact points on the clubface. An apparatus was constructed based on the pendulum principle that allowed putter golf clubs to swing at different speeds. The mean speed of the club head before ball impact, and of the ball after impact, was calculated from time measurements with photocells. A pin profile rig was used to determine the directional deviation of the golf ball. Three different putters were used in the study, two that are commercially available (toe-heel weighted and mallet types) and one specially made (wing-type) putter. The points of impact were the sweet spot (as indicated by the manufacturer's aim line), and 1, 2 and 3 cm to the left and right of the sweet spot. Calculation of club head speed before impact, and of ball speed after impact (proportional to distance), showed errors < or = 0.5% of interval duration. The variability in ball impacts was tested by measuring time and direction deviations during 50 impacts on the same ball. The mean duration (+/- s) after ball impact in the test interval (1.16 m long) was 206 (0.8) ms and the standard deviation in the perpendicular spreading of the balls in relation to the direction of the test interval was 0.005 m. A test-retest of one putter on two consecutive days after remounting of the putter on the test apparatus showed less than 1% difference in distance deviation. We conclude that the test apparatus enables a precise recording of distance and direction deviation in golf putters as well as comparisons between different putters. The apparatus and set-up can be used in the laboratory as well as outdoors on the putting green. PMID- 16368624 TI - Auditory concurrent feedback benefits on the circle performed in gymnastics. AB - In this study, we examined the effectiveness of auditory concurrent feedback on body segmental alignment during the circle movement performed on a pommel horse. Eighteen gymnasts were assigned to one of two groups: a concurrent auditory feedback group (experimental) or a control group that received no concurrent feedback. After 2 weeks of training (300 circles), the body segmental alignment (BSA) of the experimental group had improved by 2.3% between the pre test (85.7 +/- 4.8% BSA(max)) and the post test (87.7 +/- 4.0% BSA(max)). Furthermore, the results of a retention test administered 2 weeks after the post test revealed no decline in performance for the experimental group. No gains in body segmental alignment were found for the control group. It was concluded that augmented auditory feedback made available in real time can be used to correct complex movements, such as the circle movement on a pommel horse, and does not appear to lead to information-dependence despite the frequent administration of feedback. The auditory signal available in real time could help gymnasts to become more objective about their own intrinsic information necessary for the refinement of the circle movement. PMID- 16368625 TI - The measurement of competitive anxiety during balance beam performance in gymnasts. AB - The purpose of the present study was to investigate competitive anxiety during balance beam performance in gymnasts. Competitive anxiety was assessed continuously by heart rate monitoring and by retrospective self-report of nervousness in eight female national level gymnasts during their balance beam routine during one competition and two training sessions. A significant negative correlation was found between the score of the retrospective self-report of nervousness and performance during the routine. There were no significant differences in performance score by the judges between the three test sessions. There were also no differences in the retrospective self-report of nervousness. However, heart rate was significantly higher during the competition session than during the training sessions. The potential value of the retrospective report of nervousness for the study of critical events during gymnastic performance is illustrated. The results are discussed in the light of catastrophe theory. PMID- 16368626 TI - Effects of muscle-tendon length on joint moment and power during sprint starts. AB - The aim of this study was to examine the effects of muscle-tendon length on joint moment and power during maximal sprint starts. Nine male sprinters performed maximal sprint starts from the blocks that were adjusted either to 40 degrees or 65 degrees to the horizontal. Ground reaction forces were recorded at 833 Hz using a force platform and kinematic data were recorded at 200 Hz with a film camera. Joint moments and powers were analysed using kinematic and kinetic data. Muscle - tendon lengths of the medial gastrocnemius, soleus, vastus medialis, rectus femoris and biceps femoris were calculated from the set position to the end of the first single leg contact. The results indicated that block velocity (the horizontal velocity of centre of mass at the end of the block phase) was greater (P < 0.01) in the 40 degrees than in the 65 degrees block angle condition (3.39 +/- 0.23 vs. 3.30 +/- 0.21 m . s(-1)). Similarly, the initial lengths of the gastrocnemius and soleus of the front leg in the block at the beginning of force production until half way through the block phase were longer (P < 0.001) in the 40 degrees than in the 65 degrees block angle condition. The initial length and the length in the middle of the block phase were also longer in the 40 degrees than in the 65 degrees block angle condition both for both the gastrocnemius (P < 0.01) and soleus (P < 0.01-0.05) of the rear leg. In contrast, the initial lengths of the rectus femoris and vastus medialis of the front leg were longer (P < 0.05) in the 65 degrees than in the 40 degrees block angle condition. All differences gradually disappeared during the later block phase. The peak ankle joint moment (P < 0.01) and power (P < 0.05) during the block phase were greater in the 40 degrees than in the 65 degrees block angle condition for the rear leg. The peak ankle joint moment during the block phase was greater (P < 0.05) in the 40 degrees block angle for the front leg, whereas the peak knee joint moment of the rear leg was greater (P < 0.01) in the 65 degrees block angle condition. The results suggest that the longer initial muscle-tendon lengths of the gastrocnemius and soleus in the block phase at the beginning of force production contribute to the greater peak ankle joint moment and power and consequently the greater block velocity during the sprint start. PMID- 16368627 TI - The relationship between extension of the metatarsophalangeal joint and sprint time for 100 m Olympic athletes. AB - Selected kinematic variables of the foot segments and the metatarsophalangeal (MTP) joint were investigated in relation to sprinting performance among 100 m sprint athletes at the 2000 Summer Olympic Games. It was hypothesized that the kinematics of the MTP joint, and forefoot and rearfoot segments, are related to sprint performance for both male and female athletes. Kinematic sagittal plane data were collected using two digital video cameras recording at 120 fields per second. It was determined that faster male sprinters experienced higher maximal rates of MTP extension, and faster female sprinters touch down with higher posterior sole angles and take off with lower posterior sole angles. PMID- 16368628 TI - Fingertip force and electromyography of finger flexor muscles during a prolonged intermittent exercise in elite climbers and sedentary individuals. AB - The aim of this study was to characterize forearm muscle fatigue identified by the decrease in electromyogram median frequency and/or fingertip force during intermittent exercise. Nine elite climbers (international competitive level, USA 5.14a on sight) and ten non-climbers were instructed to maintain a fingertip force of 80% of their maximal voluntary contraction force on a dynamometer mimicking a rock climbing grip during a 5 s effort/5 s rest cycle for 36 repetitions (i.e. 6 min of exercise). Elite climbers lasted twice as long as non climbers (climbers: 3 min; non-climbers: 1 min 30 s) before the force could no longer be maintained (i.e. the failure point). After this moment, fingertip force decreased and stabilized until the end of the exercise around 50% maximum voluntary contraction force in non-climbers and 63% in elite climbers. Electromyogram median frequency showed a greater decrease in non-climbers than in elite climbers before the failure point. No change in median frequency was observed after the failure point in elite climbers or in non-climbers. These results confirm that elite climbers are better adapted than non-climbers for performing the intermittent fingertip effort before the failure point. After this point, the better fingertip force of elite climbers suggests different forearm muscle properties, while the electromyography results do not provide any indication about the fatigue process. PMID- 16368629 TI - The influence of rhythm and personality in the endurance response to motivational asynchronous music. AB - In this study, we examined participants' responses to motivational asynchronous music by isolating rhythmical properties and exploring personality correlates. Fifty-eight physically active participants (41 men and 17 women) aged 22.3 +/- 6.4 years performed an isometric weight-holding task on three occasions while being randomly exposed to no music, rhythm and motivational music. The rhythm and music conditions were edited portions of the same musical selection and had identical fast tempi, although the rhythm condition contained no melody, harmonies or lyrics. Participants each completed a copy of Cattell's 16PF following the third and final trial. A repeated-measures analysis of variance found the participants held the weight suspended for significantly longer when listening to motivational music in comparison to rhythm or no music. When listening to rhythm, participants endured the task for significantly longer than when listening to no music. The response to music was found to be significantly related to liveliness, while sensitivity correlated with responses to music factors (harmony, melody, lyrics, etc.) not present in the rhythm condition. These results suggest that responses to motivational music are subtle in nature and are determined by both musical factors and individual characteristics, and potentially an interaction between the two. PMID- 16368630 TI - Movement and stretching imagery during flexibility training. AB - The aim of this study was to examine the effect of movement and stretching imagery on increases in flexibility. Thirty volunteers took part in a 4 week flexibility training programme. They were randomly assigned to one of three groups: (1) movement imagery, where participants imagined moving the limb they were stretching; (2) stretching imagery, where participants imagined the physiological processes involved in stretching the muscle; and (3) control, where participants did not engage in mental imagery. Active and passive range of motion around the hip was assessed before and after the programme. Participants provided specific ratings of vividness and comfort throughout the programme. Results showed significant increases in flexibility over time, but no differences between the three groups. A significant relationship was found, however, between improved flexibility and vividness ratings in the movement imagery group. Furthermore, both imagery groups scored significantly higher than the control group on levels of comfort, with the movement imagery group also scoring significantly higher than the stretching imagery group. We conclude that the imagery had stronger psychological than physiological effects, but that there is potential for enhancing physiological effects by maximizing imagery vividness, particularly for movement imagery. PMID- 16368631 TI - The convergent and discriminant validity of burnout measures in sport: a multi trait/multi-method analysis. AB - Athlete burnout research has been hampered by the lack of an adequate measurement tool. The Athlete Burnout Questionnaire (ABQ) and the Maslach Burnout Inventory General Survey (MBI-GS) are two recently developed self-report instruments designed to assess burnout. The convergent and discriminant validity of the ABQ and MBI-GS were assessed through multi-trait/multi-method analysis with a sporting population. Overall, the ABQ and the MBI-GS displayed acceptable convergent validity with matching subscales highly correlated, and satisfactory internal discriminant validity with lower correlations between non-matching subscales. Both scales also indicated an adequate discrimination between the concepts of burnout and depression. These findings add support to previous findings in non-sporting populations that depression and burnout are separate constructs. Based on the psychometric results, construct validity analysis and practical considerations, the results support the use of the ABQ to assess athlete burnout. PMID- 16368632 TI - The relationship between peak height velocity and physical performance in youth soccer players. AB - Longitudinal changes in height, weight and physical performance were studied in 33 Flemish male youth soccer players from the Ghent Youth Soccer Project. The players' ages at the start of the study ranged from 10.4 to 13.7 years, with a mean age of 12.2 +/- 0.7 years. Longitudinal changes were studied over a 5 year period. Peak height velocity and peak weight velocity were determined using non smoothed polynomials. The estimations of peak height velocity, peak weight velocity and age at peak height velocity were 9.7 +/- 1.5 cm x year-1, 8.4 +/- 3.0 kg x year-1 and 13.8 +/- 0.8 years, respectively. Peak weight velocity occurred, on average, at the same age as peak height velocity. Balance, speed of limb movement, trunk strength, upper-body muscular endurance, explosive strength, running speed and agility, cardiorespiratory endurance and anaerobic capacity showed peak development at peak height velocity. A plateau in the velocity curves was observed after peak height velocity for upper-body muscular endurance, explosive strength and running speed. Flexibility exhibited peak development during the tear after peak height velocity. Trainers and coaches should be aware of the individual characteristics of the adolescent growth spurt and the training load should also be individualized at this time. PMID- 16368633 TI - Worldwide regional variations in home advantage in association football. AB - Home advantage plays an important part in determining the result of a game of football. Its existence and magnitude is well documented in England, but its causes are still not completely understood. In this study, reliable estimates of home advantage are calculated for the domestic leagues of all countries of Europe and South America, as well as a selection of countries from other continents. The results of all games during the last six seasons are used for each of these 72 countries. In Europe, home advantage in the Balkan countries, especially Bosnia and Albania, is much higher than average. It is generally lower than average in northern Europe, from the Baltic republics, through Scandinavia to the British Isles. In South America, home advantage is high in the Andean countries and lower elsewhere, especially in Uruguay. Home advantage is not unusually high or low in any of the countries from other continents. A multiple regression model for the 51 European countries, which included variables for geographical location, crowd effects and travel, accounted for 76.7% of the variability in home advantage. The large geographical variations can be interpreted in terms of territoriality being a contributing factor to home advantage. PMID- 16368634 TI - Pedometer reliability, validity and daily activity targets among 10- to 15-year old boys. AB - The aims of this study were to: (1) determine whether the number of pedometer counts recorded by adolescents differs according to the adiposity of the participant or location on the body; (2) assess the accuracy and reliability of pedometers during field activity; and (3) set adolescent pedometer-based physical activity targets. Seventy-eight 11- to 15-year-old Boy Scouts completed three types of activity: walking, fast walking and running. Each type was performed twice. Participants wore three pedometers and one activity monitor during all activities. Participants were divided into groups of normal weight (BMI < 85th percentile) and at risk of being overweight (BMI > or = 85th percentile). Intra class correlations across the three activities indicated reliability (r = 0.51 - 0.92, P < 0.001). This conclusion was supported by narrow limits of agreement that were within a pre-set range that was practically meaningful. Multivariate analysis of covariance indicated adiposity group differences, but this difference was a function of the increased stature among the larger participants (P < 0.001). Ordinary least-squares regression models and multi-level regression models showed positive associations between the number of pedometer and activity monitor counts recorded by the three groups of participants during all activities (all P < 0.001). The mean number of counts recorded for all participants during the fast walk was 127 counts per minute. In conclusion, the pedometers provided an accurate assessment of adolescent physical activity, and a conservative estimate of 8000 pedometer counts in 60 min is equivalent to 60 min of moderate to vigorous physical activity. PMID- 16368635 TI - Coping strategies and the influence of perceived parental support in junior national age swimmers. AB - This study examines how national junior age group swimmers cope with poor performance and explores whether coping strategy use changes with perceptions of parental support. One hundred and four junior age group swimmers (mean age 14.2 years) completed a modified version of the COPE scale (Crocker & Issack, 1997) and the perceptions of Perceived Parental Support Scale (Van Yperen, 1995). Results showed that the swimmers in this sample used a mixture of behavioural and cognitive strategies with an emphasis upon individual or internally focused approaches. When analysing coping strategies and perceptions of parental support (high, medium and low), significant differences were found in the use of behavioural and cognitively orientated approaches. When perception of parental support was low, swimmers reported using less active and training-orientated mechanisms and a tendency to use strategies such as self-blame and venting of emotion. These findings suggest that the effectiveness and ability of coping strategies to adapt to differing situations may be contingent upon perceptions of perceived support for junior swimmers. This highlights an area for future research. PMID- 16368636 TI - Imagery use by injured athletes: a qualitative analysis. AB - The purpose of this study was to expand our knowledge and increase our understanding of imagery use by athletes in sport-injury rehabilitation using a qualitative approach. The participants were 10 injured athletes who were receiving physiotherapy at the time they were interviewed. During the interviews, the athletes provided extensive information about their use of imagery during injury rehabilitation and it was clear that they believed imagery served cognitive, motivational and healing purposes in effectively rehabilitating an injury. Cognitive imagery was used to learn and properly perform the rehabilitation exercises. They employed motivational imagery for goal setting (e.g. imagined being fully recovered) and to enhance mental toughness, help maintain concentration and foster a positive attitude. Imagery was used to manage pain. The methods they employed for controlling pain included using imagery to practise dealing with expected pain, using imagery as a distraction, imagining the pain dispersing, and using imagery to block the pain. With respect to what they imaged (i.e. the content of their imagery), they employed both visual and kinaesthetic imagery and their images tended to be positive and accurate. It was concluded that the implementation of imagery alongside physical rehabilitation should enhance the rehabilitation experience and, therefore, facilitate the recovery rates of injured athletes. Moreover, it was recommended that those responsible for the treatment of injured athletes (e.g. medical doctors, physiotherapists) should understand the benefits of imagery in athletic injury rehabilitation, since it is these practitioners who are in the best position to encourage injured athletes to use imagery. PMID- 16368637 TI - The effects of self-handicapping on attributions and perceived judo competence. AB - The aim of this study was to examine hypotheses derived from Jones and Berglas's (1978) self-handicapping model. It was hypothesized that individuals using many self-handicaps would use more internal attributions and report greater gains in perceived judo ability following success than individuals using few self handicaps. In addition, it was hypothesized that individuals using many self handicaps would use more external attributions and report less reduction in perceived judo ability following failure. Fifty-three judo players completed measures of trait self-handicapping, situational self-handicapping and a measure of perceived judo ability before competition. Following competition, the participants completed the Causal Dimension Scale II and the measure of perceived judo ability for a second time. Analyses of variance revealed that high self handicappers attributed failure to more external factors than low self handicappers. It was also found that high self-handicappers reported less of a reduction in perceived judo ability following failure than low self-handicappers. The findings therefore provide support for the potential short-term benefits of self-handicapping in sport, although further research is required to examine the long-term implications of using self-handicaps. PMID- 16368638 TI - An examination of multiple goals in children's physical education: motivational effects of goal profiles and the role of perceived climate in multiple goal development. AB - In this paper, I present data from two studies that sought to examine multiple achievement goals in the context of children's physical education (PE). Study 1 examined the links that multiple-goal profiles (i.e. mastery/performance approach/performance-avoidance goals) for PE had with self-determined motivation, affective patterns and levels of extracurricular sporting activity in a sample of 193 Year 7 pupils. Results suggested that children endorsing high mastery/high performance-approach/high performance-avoidance, high mastery/high performance approach/low performance-avoidance, or high mastery/ low performance-approach/low performance-avoidance profiles typically exhibited the most adaptive motivational responses. In contrast, children endorsing a low mastery/high performance approach/high performance-avoidance profile typically experienced more maladaptive motivational outcomes. Study 2 examined the development of multiple achievement goals over two terms of PE in conjunction with retrospective perceptions of the motivational climate in the sample. The results of analysis of variance suggested that children exposed to a consistent high mastery/low performance climate experienced decreased performance-avoidance goals and maintained high levels of mastery goals for PE. In contrast, children exposed to a consistent low mastery/high performance climate experienced increased performance-avoidance goals and decreased mastery goals for PE. PMID- 16368639 TI - Approach and avoidance coping during task performance in young men: the role of goal attainment expectancies. AB - The present study investigated approach and avoidance coping strategies as a function of goal attainment expectancies in young men. Twenty-four male students participated in an experiment consisting of two rowing tests. After the initial test, participants were divided into two experimental groups and were assigned to conditions of attainable and unattainable goals for the final test, depending on performance in the initial test and experimental condition. Participants of the two groups reported similar goal attainment expectancies before the start of the final test; however, during the task expectancies for the attainable goal group increased, whereas those for the unattainable goal group decreased. After the conclusion of the task, participants were asked to respond with regard to coping strategies they used during the latest part of the final test, when goal attainment expectancies had been differentiated. The analysis revealed significant effects for effort increase, and mental and behavioural disengagement, whereas a non-significant but considerable effect was identified for planning. The higher goal attainment expectancies group scored higher on effort increase and planning, and lower on behavioural and mental disengagement, than the lower goal attainment expectancies group. The results of the present study are in accordance with the general theoretical assumptions of the control process theory, and more particularly highlight the importance of goal attainment expectancies as a factor influencing the employment of approach and avoidance coping strategies in achievement situations. PMID- 16368640 TI - Expertise and talent development in rugby refereeing: an ethnographic enquiry. AB - We explore how expertise is obtained in the domain of rugby refereeing. The research data are qualitative and are drawn from an 18 month period working in collaboration with the Rugby Football Union Elite Referee Unit. Adopting an ethnographic mode of enquiry, the study combined long-term participant observation with in-depth interviewing, indirect observations and the collection of artefacts including existing protocol, coach feedback forms and strategic reports. The diversity of methodologies allowed us to examine how expertise is developed across various domains of analysis, including the intrapersonal, interpersonal, group and social perspectives. Building on expertise studies in "deliberate practice", further prerequisites for expertise, at least in this domain and with these participants, incorporated "deliberate experience" and "transfer of skills". Additionally, a key issue in the findings concerns a shift from "descriptive" towards a "non-linear processes"-oriented model of development. We conclude by identifying opportunities and limitations associated with the adoption of ethnography as a method for studying expertise. PMID- 16368641 TI - A distributional analysis of the effect of physical exercise on a choice reaction time task. AB - The aim of this study was to examine the facilitating effects of physical exercise on the reaction process. Eleven participants with specific expertise in decision-making sports performed a choice reaction time task during moderate sub maximal exercise (90% of their ventilatory threshold power). Participants were tested at rest and while cycling. During exercise, the participants were faster, without being more variable. We suggest that the effect of exercise on cognitive performance was due to a major generalized improvement of the whole distribution of response time and, although the benefit effect was small, it was consistent throughout the entire range of reaction times. PMID- 16368642 TI - Carcinogenicity of radon/radon decay product inhalation in rats--effect of dose, dose rate and unattached fraction. AB - PURPOSE: The effects of inhalation of radon/radon decay products at different total doses, dose rates and 'unattached' fractions were investigated in a life span study in rats. MATERIALS AND METHODS: 1574 rats inhaled radon/radon decay products in a purpose-built recirculating exposure system that provided stable/reproducible exposure conditions. 501 were maintained as controls. RESULTS: Lung tumour incidences were significantly elevated in most exposed groups. The study power was insufficient to resolve the shape of the dose and dose rate response curves, but combination of this data with that from other studies demonstrated that for high cumulative exposures, the lifetime excess absolute risk increases with increasing exposure durations and for low cumulative exposures the opposite trend occurs. Exposure did not increase leukaemia incidences. A small number of non-lung tumour types including mammary fibroadenoma showed elevated incidences in some exposed groups, however not consistently across all exposure groups and showed no dose or dose rate relationship. CONCLUSIONS: Radon/radon decay product exposure caused excess lung tumours in rats along with limited non-lung effects. The results are consistent with the findings that at low cumulative exposures decreasing exposure concentrations or protracting the time over which the dose is delivered, reduces lung tumour risk. At higher levels, decreasing exposure concentrations or protracting exposure time increases lung tumour risk. PMID- 16368643 TI - Radiation tolerance in the eutardigrade Richtersius coronifer. AB - PURPOSE: Tardigrades have a reputation of being extremely tolerant to extreme environmental conditions including tolerance to ionizing radiation while in a desiccated, anhydrobiotic state. However, the evidence for radio-tolerance in tardigrades is based on only one previous report, and there is an obvious need for complementary studies. In this paper we report an investigation on radio tolerance in desiccated and hydrated specimens of the eutardigrade Richtersius coronifer. MATERIALS AND METHODS: Groups of 30 - 50 tardigrades were exposed to gamma-radiation at doses between 1.0 - 9.0 (anhydrobiotic animals) or 0.5 - 5.0 (hydrated animals) kGy and the animals were followed until all were dead. Radiation tolerance of both desiccated and hydrated tardigrades was studied. RESULTS: Both desiccated and hydrated animals irradiated with 0.5 and 1 kGy did not deviate in survival from the control groups. Animals from all exposed groups underwent their moulting and egg production cycle, but at decreasing frequency for doses above 1 kGy. No eggs laid by irradiated animals hatched, while eggs laid by controls did so. CONCLUSION: Our study suggests that radiation tolerance in tardigrades is not due to biochemical protectants connected with the desiccated state. Rather, cryptobiotic tardigrades may rely on efficient mechanisms of DNA repair, the nature of which is currently unknown. PMID- 16368644 TI - Radiation treatment of acute inflammation in mice. AB - PURPOSE: Low-dose radiotherapy (RT) has often been used effectively for the treatment of a variety of benign diseases, particularly those with acute inflammatory features. Here we report findings on radiation treatment of acute inflammation using a murine carrageenin air pouch model. MATERIALS AND METHODS: Air pouches raised on the dorsal surface of mice were injected with lambda carrageenin and were irradiated 6 h later with doses ranging from 0-5 Gy. Treatment success was evaluated at various times thereafter by volume of exudate and number of inflammatory cells, and levels of inflammation-related cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1beta) and transforming growth factor beta-1 (TGFbeta-1), and expression of inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), cyclooxygenase-2 (COX-2) and inducible heat shock protein 70 (HSP70) as determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting, respectively. RESULTS: Crude inflammatory parameters such as the amount of exudates and number of inflammatory cells remained largely unaffected by radiation or were even a slightly and transiently increased. However, the expression of iNOS was attenuated by radiation concomitant with an increase in the levels of HO-1 and HSP70. Cytokine levels varied with the radiation dose and the time point. CONCLUSIONS: Ionizing radiation, even at low doses, functionally modulates inflammatory cells. Our findings indicate possible mechanisms as to how low-dose radiation may exert anti inflammatory effects and provide the first evidence that heat shock proteins may be involved in this response. PMID- 16368645 TI - Effects of gamma radiation on luteinizing hormone (LH) receptor expression, signal transduction and steroidogenesis in cultured rat Leydig cells. AB - PURPOSE: To investigate the dose-dependent effects of gamma radiation on rat Leydig cell luteinizing hormone (LH) receptor expression, signal transduction and steroidogenesis. MATERIALS AND METHODS: Leydig cells isolated from testicles of adult male albino rats were cultured and exposed to different doses (2, 4, 6, 8 and 10 Gy) of fractioned gamma radiation for three days. Normal as well as irradiated cells were used for the quantification of cell surface LH receptor or LH/cyclic adenosine monophosphate (cAMP) stimulation test or total ribonucleic acid (RNA) isolation to study the LH receptor messenger RNA (mRNA) expression by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Radiation was found to have dose-dependent effects on rat Leydig cell surface LH receptor, basal and LH-stimulated cAMP production, basal, LH and cAMP-stimulated steroidogenesis. Lower doses (2 and 4 Gy) of radiation did not induce any change, whereas higher doses (6, 8 and 10 Gy) of radiation exposure diminished the above said parameters significantly. In spite of the low level of cell surface LH receptors following higher doses of radiation exposure, its mRNA level was not altered under any dose level. CONCLUSION: From these observations, it is concluded that gamma radiation has dose-dependent inhibitory effects on Leydig cell LH signal transduction and steroidogenesis. Decreased cell surface LH receptor following higher doses of radiation exposure is not coupled with altered LH receptor mRNA level. PMID- 16368646 TI - Induction of radical scavenging ability and suppression of lipid peroxidation in rat liver microsomes following whole-body, low-dose X-irradiation. AB - PURPOSE: To investigate changes in radical scavenging ability and lipid peroxidation in liver microsomal membranes and cooperative suppression of lipid peroxidation by microsomal and cytosolic radical scavengers, 24 h after whole body, low-dose X-irradiation of rats. MATERIALS AND METHODS: Male Wistar rats were irradiated with 1-50 cGy of X-rays. Liver microsomal radical scavenging ability was determined using the trapping ability of 1,1-diphenyl-2 picrylhydrazyl (DPPH), a stable free radical. Microsomal alpha-tocopherol (Vit.E) content was determined using an electrochemical detector. Microsomal glutathione peroxidase (GPx) activity was determined as the consuming rate of NADPH. Microsomal lipid peroxidation was determined by the thiobarbituric acid method. RESULTS: Low molecular weight radical scavenging ability of rat liver microsomes increased 24 h after whole-body, low-dose X-irradiation when alpha-tocopherol was included, showing a maximum level at 5-10 cGy. Microsomal GPx activity also increased 24 h after 5 cGy irradiation. The lipid peroxidation level in microsomes decreased, showing a maximal suppression at 5 cGy. High-dose irradiation-induced microsomal lipid peroxidation was strongly suppressed cooperatively by microsomal and cytosolic antioxidants induced by low-dose irradiation. CONCLUSION: Low doses of radiation induce increases in liver microsomal antioxidants, which in turn result in enhanced suppression of microsomal lipid peroxidation cooperatively with cytosolic antioxidants induced by low-dose irradiation. PMID- 16368647 TI - Dose rate dependence of the relative biological effectiveness of 103Pd for continuous low dose rate irradiation of BA1112 rhabdomyosarcoma cells in vitro relative to acute exposures. AB - PURPOSE: To measure the relative biological effectiveness (RBE) of continuous low dose rate irradiation (CLDRI) using 103Pd sources relative to acute high dose rate irradiations (AHDRI) from a 250 kVp x-ray beam and an x-ray beam having an equivalent mono-energetic photon energy equal to the average energy of the 103Pd source for BA1112 rhabdomyosarcoma cells. MATERIALS AND METHODS: A customized 103Pd irradiator was built to provide CLDRI using 103Pd at different dose rates relevant to clinical interstitial brachytherapy to BA1112 rhabdomyosarcoma cells growing in exponential phase in culture. A special x-ray beam that simulates the photon energies emitted by the 103Pd source was also developed to provide acute high dose rate irradiation at those energies. Cell survival curves from different irradiation conditions were measured. The RBE with respect to AHDRI using standard 250 kVp x-rays was determined from the doses required to achieve a cell surviving faction of 0.01. RESULTS: For acute irradiation, the RBE of the x-rays simulating (103)Pd was 1.24 relative to 250 kVp x-rays. A profound dose rate effect was observed at low dose rates in the range of 6.8 - 14.4 cGy/h that are typical of permanent interstitial brachytherapy. At cell-surviving fraction of 0.01, the RBE of CLDRI at 6.8 and 14.4 cGy/h using 103Pd sources was reduced by a factor of 3 and 2, respectively, relative to the acute exposure. This observation is in good agreement with recent in vivo tumor cure studies performed on BA1112 tumor. CONCLUSION: The relative biological effectiveness of the photons emitted by 103Pd depends on both the linear energy transfer (LET) of the low energy photons and the dose rate of the irradiation. The higher LET of 103Pd photons is biologically more effective in killing BA1112 tumor cells compared to conventional 250 kVp x-rays when both are delivered at the same dose rate. But the gain in RBE that results from the higher LET can be quickly negated by the reduced dose rate of the irradiation. PMID- 16368648 TI - Increased expression of GRP94 protein is associated with decreased sensitivity to X-rays in cervical cancer cell lines. AB - PURPOSE: Radiation therapy is one of the standard treatments for cervical cancer. Glucose regulated protein 94 (GRP94) is a molecular chaperone, which increases in amount after X-ray irradiation. This study examined the involvement of GRP94 in radio-resistance in human cervical cancer cells. MATERIALS AND METHODS: Seven human cervical carcinoma cell lines (HeLa, SKG-I, SKG-IIIb, QG-U, Caski, SiHa and C33A) were examined for basal levels of GRP94 protein by western blotting analysis. Sensitivity to X-ray irradiation of these cell lines was determined with a colony survival assay. The suppression of GRP94 expression was performed using specific small-interfering RNA (siRNA) in HeLa and Caski cells. RESULTS: HeLa cells and QG-U cells, with higher basal levels of GRP94, exhibited a low sensitivity to X-ray cell killing. In HeLa cells, the sensitivity increased when protein GRP94 levels were reduced by specific siRNA transfection. However, a reduction in GRP94 protein had little effect on the X-ray sensitivity of Caski cells, which expressed low basal GRP94 protein levels but showed a low sensitivity to X-rays. CONCLUSIONS: High basal protein levels of GRP94 were correlated with a modest decrease in sensitivity to X-ray cell death in some cervical cancer cell lines. These results suggest that higher GRP94 protein expression is one of the molecular mechanisms causing resistance to radiation, and therefore GRP94 siRNA might be useful in tumor-specific gene therapy by reversing radio-resistance prior to radiation in cervical cancer. PMID- 16368649 TI - Attenuation of chronic thermotolerance by KNK437, a benzylidene lactam compound, enhances thermal radiosensitization in mild temperature hyperthermia combined with low dose-rate irradiation. AB - PURPOSE: We investigated whether the attenuation of chronic thermotolerance by KNK437, a heat shock protein inhibitor, can modify the effect of thermal radiosensitization in mild temperature hyperthermia (MTH) combined with low dose rate irradiation (LDRI). MATERIALS AND METHODS: The human lung adenocarcinoma cell line A549 was simultaneously exposed to LDRI with MTH at 41 degrees C and KNK437 at a dose of 100 microM. Cell survival was estimated by a clonogenic assay. Cell cycle change during treatment was analyzed by flow cytometry. Expression levels of the heat shock proteins hsp72, hsp27 and heat shock factor 1 (HSF-1) were measured by Western blotting. RESULTS: KNK437 inhibited the expression of inducible hsp72 and hsp27, but produced no change in the mobility shift of HSF-1. The cytotoxicity of LDRI was enhanced by MTH. The survival curve for LDRI + MTH revealed no development of chronic thermotolerance up to 48 h. Simultaneous LDRI and KNK437 treatment also resulted in enhanced cell killing. The radiosensitizing effect of KNK437 was enhanced by simultaneous exposure of the cells to MTH. Flow cytometry analysis of cell cycle progression demonstrated marked G2 arrest and mild G1 arrest with LDRI alone, but mild G1 arrest with MTH alone, and mild G2-M, S-phase accumulation with KNK437 alone. The marked G2 arrest caused by LDRI was partially suppressed by the addition of MTH, and was also suppressed by KNK437 treatment. CONCLUSIONS: Exposure of A549 cells to KNK437 caused inhibition of hsp72 and hsp27 expression. The addition of KNK437 increased not only thermosensitivity to MTH, but also radiosensitivity to LDRI. KNK437 also enhanced the MTH-induced radiosensitization under these experimental conditions. PMID- 16368650 TI - Immunohistochemical study of tumor necrosis factor-alpha, matrix metalloproteinase-12, and tissue inhibitor of metalloproteinase-2 on alveolar macrophages of BALB/c mice exposed to short-term cigarette smoke. AB - Alveolar cells were evaluated in BALB/c mice exposed to smoke from 9 cigarettes per day. The mice were sacrificed at 1, 5, and 10 days and examined by lung morphometry and immunohistochemical staining of alveolar macrophages for tumor necrosis factor (TNF)-alpha, matrix metalloproteinase (MMP)-12, and tissue inhibitor of metalloproteinase (TIMP)-2. Cigarette smoke (CS)-exposed mice showed a progressive increase in numbers of alveolar macrophages (AMs) up to 10 days. MMP-12 was increased in AMs compared to the control group. The number of AMs with TNF-alpha and TIMP-2 labeling was higher at 5 days but decreased at 10 days. Short-term CS exposure induced pulmonary inflammation with major participation of MMP-12-positive AMs. PMID- 16368651 TI - Expression of angiostatin, integrin alphavbeta3, and vitronectin in human lungs in sepsis. AB - Angiostatin, integrin alphavbeta3, and vitronectin play important roles in inflammation. However, there is very little information on expression of these molecules in the lungs of humans with sepsis. Therefore, as a first step to eventually study the function of these molecules, the authors conducted an immunohistochemical study to evaluate their expression in lungs of normal (N = 8) and sepsis patients (N = 8). In normal lungs, angiostatin expression was minimal in the alveolar septa and alveolar macrophages, and absent in large blood vessels, bronchioles, and interstitium. In sepsis patients, the staining was intense in the septa, neutrophils, alveolar macrophages, and large blood vessels. Integrin alphavbeta3 staining was observed in occasional bronchiolar epithelial cells and a few alveolar macrophages in the normal lungs. The integrin was expressed extensively and intensely in bronchiolar epithelium and alveolar macrophages, and with lesser intensity in large blood vessels in inflamed lungs. Compared to the normal lung, vitronectin expression was increased in alveolar macrophages and in vascular smooth muscles in inflamed lungs. These data show cell-specific increase in the expression of integrin alphavbeta3, angiostatin, and vitronectin in inflamed lungs of sepsis patients. Because all these molecules can have significant influence on inflammation, the data reported in this manuscript create a need for further investigation. PMID- 16368653 TI - Prescription drug abuse among ecstasy users in Miami. AB - This study examines the nature, extent and consequences of prescription drug abuse among 143 ecstasy users in Miami. Participants were recruited through nightclub and college campus outreach, and through respondent referrals. Instrumentation included the Risk Behavior Assessment, Substance Abuse Module and Center for Epidemiological Studies Depression Scale. Median age was 23, 42% were female and 50% Hispanic. An arrest history was reported by 44%, and 33% reported prior drug/alcohol treatment. Prescription drug abuse was reported by 87%; alprazolam (57%), oxycodone (36%), hydrocodone (32%) and diazepam (30%) were cited most often. Prescription drug abusers were more likely to report polydrug use, drug treatment histories, risky drug use behaviors, and symptoms of depression. They also reported numerous physical, psychological and social consequences of prescription drug abuse. Additional studies among larger samples are needed to understand the processes of prescription drug access and the extent of integration in club drug using cultures. PMID- 16368652 TI - Divergent effects of BMP-2 on gene expression in pulmonary artery smooth muscle cells from normal subjects and patients with idiopathic pulmonary arterial hypertension. AB - Bone morphogenetic proteins (BMPs) inhibit proliferation and induce apoptosis in pulmonary artery smooth muscle cells (PASMCs) from normal subjects. Dysfunction of BMP signaling due to mutations in and/or down-regulation of BMP receptors has been implicated in idiopathic pulmonary arterial hypertension (IPAH). The authors examined whether BMP differentially regulates gene expression in PASMCs from normal subjects and IPAH patients using the Affymetrix microarray analysis. BMP-2 treatment (200 nM for 24 hours) altered expression levels of 6206 genes in normal and IPAH PASMCs. Of these genes, 1063 were regulated oppositely by BMP-2: 523 genes were down-regulated by BMP-2 in normal PASMCs but up-regulated in IPAH PASMCs, whereas 540 genes were up-regulated by BMP-2 in normal PASMCs but down regulated in IPAH PASMCs. The divergent effects of BMP-2 on gene expression profiles indicate that PASMCs may undergo significant phenotypic changes in IPAH patients during development of the disease. The transition of the antiproliferative effect of BMP-2 in normal PASMCs to its proliferative effect in IPAH patients is attributed potentially to its differential effect on expression patterns of various genes that are involved in cell proliferation and apoptosis. Among the 6206 BMP-2-sensitive genes, there are more than 1800 genes whose expression levels were negatively (correlation coefficient, r, <-0.9) or positively (with r >+ 0.9) correlated with the pulmonary arterial pressure. These results suggest that BMP-mediated gene regulation is significantly altered in PASMCs from IPAH patients and mRNA expression changes in BMP-regulated genes may be involved in the development of IPAH. PMID- 16368654 TI - Co-consumption of benzodiazepines in heroin users, methadone-substituted and codeine-substituted patients. AB - Concomitant consumption of benzodiazepines (BZDs) and opioids is a major problem in patients with opioid dependence. It may have substantial impact on morbidity, mortality and clinical course. The current retrospective study aims to determine whether there are differences in the additional use of BZDs among addicts regularly taking methadone or codeine medications in treatment and untreated addicts injecting heroin. The records of 1,685 patients admitted for detoxification were analyzed using bivariate analysis and multiple logistic regression analysis. Demographic and drug related variables were considered, both as possible confounders and predictors of concomitant BZD use. Daily intake of BZDs was reported in 44.4% of the patients. Patients treated with methadone or codeine medications report daily intake of BZDs significantly more often than the heroin-dependent patients (p < 0.01). Using multiple regression analyses, the results were confirmed as independent from the assessed possible confounders. Further we found that daily use of alcohol or barbiturates, early onset of opioid use (p < 0.01), unemployment, having a substance dependent family member with, and a history of imprisonment (for all p < 0.05) were associated with concomitant daily consumption of BZDs in opioid dependent subjects. These finding underline the need to further explore the causes, interactions and consequences of concomitant BZD and opiate use. PMID- 16368655 TI - Toxicology screening in orthopedic trauma patients predicting duration of prescription opioid use. AB - Following hospitalization for orthopedic trauma, some patients continue to use opioids following fracture healing. This retrospective cohort study of 50 patients with high-energy fractures was conducted to determine if toxicology screening tests upon admission can predict subsequent opioid use. Data were collected from clinical records and a statewide electronic database of prescription records. Six months following hospital discharge, those with positive toxicology used more Following hospitalization for orthopedic trauma, some patients continue to use opioids following fracture healing. This retrospective cohort study of 50 patients with high-energy fractures was conducted to determine if toxicology screening tests upon admission can predict subsequent opioid use. Data were collected from clinical records and a statewide electronic database of prescription records. Six months following hospital discharge, those with positive toxicology used more. Following hospitalization for orthopedic trauma, some patients continue to use opioids following fracture healing. This retrospective cohort study of 50 patients with high-energy fractures was conducted to determine if toxicology screening tests upon admission can predict subsequent opioid use. Data were collected from clinical records and a statewide electronic database of prescription records. Six months following hospital discharge, those with positive toxicology used more opioids (730 mg vs. 364 mg; P = .04) expressed as morphine equivalents than those with negative toxicology and were more likely to continue using opiates at the end of the 3rd, 4th, 5th, and 6th month after discharge. Patients hospitalized for high-energy fractures with positive admission toxicology are at risk for prolonged opiate use during the initial six months following discharge. PMID- 16368656 TI - Racial and ethnic diversity among a heroin and cocaine using population: treatment system utilization. AB - Knowledge about the meanings and consequences of behaviors associated with drug use among diverse populations is essential for developing effective public health and clinical strategies. In this study we identify racial/ethnic variations in patterns of drug use, Addiction Severity Index (ASI) scores, response to intervention, concordance between self-report of drug use and biochemical confirmation, and treatment system contacts in a sample of 1175 out-of-treatment cocaine and heroin users drawn from a trial of brief motivation in the outpatient clinics of an inner-city academic hospital. Key differences were identified in drug of choice, in all of the ASI domains except medical, in validity of self report of use, and in rate of treatment contact. Differences related to race and ethnicity should be evaluated to determine needs for a variety of substance abuse treatment modalities, assure timely access to culturally competent care, and develop policies that are tailored to real conditions. PMID- 16368657 TI - Symptom endorsement of alcohol and other substance abuse and dependence criteria among American-Indian adolescents in residential substance abuse treatment. AB - OBJECTIVE: This study addresses the prevalence of DSM-IV substance abuse and dependence and the endorsement of specific symptoms of these disorders among American Indian adolescents admitted to a residential substance abuse treatment program. METHOD: We interviewed 89 American Indian adolescents using the Composite International Diagnostic Interview-Substance Abuse Module (CIDI-SAM). RESULTS: The most frequent diagnoses included marijuana dependence (51.7%), followed by alcohol abuse and dependence (33.7% each) and marijuana abuse (32.6%). Among the three substances with the highest prevalence of any use disorder (alcohol, marijuana and stimulants), the most frequently endorsed abuse criterion was Impaired Role Obligations. The two most frequently endorsed dependence criteria were Use Despite Substance-Related Psychological/Physical Problems and Unsuccessful Attempts to Quit/Cut down on substance use. CONCLUSIONS: These American Indian adolescents presented to a residential substance abuse treatment program with serious, highly complex substance use disorders that represent substantial challenges to effective treatment. PMID- 16368658 TI - Dual pharmacotherapy and motivational interviewing for tobacco dependence among drug treatment patients. AB - This pilot study is the first to examine the feasibility and outcomes of dual pharmacotherapy for smoking cessation among drug treatment patients. The intervention consisted of 7 weeks of bupropion (300 mg), 12 weeks of nicotine gum, and 6 sessions of motivational interviewing. The trial was conducted among 28 patients recruited from 5 methadone clinics and employed a pretest-posttest design. At 6 months post quit date, 14% of participants met criteria for biochemically-verified abstinence. Among those still smoking, number of cigarettes smoked decreased significantly and most (88%) had made at least 1 serious quit attempt. Participation rates were excellent and no adverse effects on alcohol or illicit drug use were found. Although not a definitive test of the intervention, findings suggest that a multi-component approach to tobacco dependence is feasible and potentially effective in helping drug treatment patients achieve smoking cessation well beyond the end of treatment and that a large-scale randomized trial is warranted. PMID- 16368659 TI - The relation between social desirability and different measures of heroin craving. AB - A low association between self-reported craving and physiological measures of craving is often found. Social desirability might influence this relation between subjective and physiological reactivity. Subjects were seventy-six in-patient abstinent heroin abusers. Social desirability, general craving scores and difference scores of physiological and subjective heroin craving, measured after exposure to a neutral and heroin cue were assessed. Cue reactivity, both subjective and physiological was found to be enhanced after exposure to a heroin related cue. In line with earlier studies, a modest relation was found between subjective and physiological cue reactivity. Contrary to earlier research, however, high social desirability scores were associated with lower craving scores, but not with physiological scores. It can be concluded that, among other subject-related variables, social desirability influences self-reported craving but does not influence physiological reactivity in a clinical population. No significant moderation effect of social desirability on the relation between subjective and physiological reactivity was found. PMID- 16368660 TI - Drug counselor report of adolescents abuse of nicotine replacement therapy. AB - BACKGROUND: Nicotine replacement products (NRT) are formulated and marketed to reduce their abuse liability among adolescents. Few studies have examined the extent of adolescent abuse. The objective of this manuscript is to describe the youth abuse rate for NRT and other over-the-counter (OTC) abusable substances. METHODS: Two cross-sectional telephone surveys of Safe and Drug Free School Coordinators were conducted in 1996/7 (N = 562) and 1998/9 (N = 501). Abuse of NRT and other OTC drugs and circumstances surrounding NRT abuse was ascertained. RESULTS: NRT abuse rates were low and did not change significantly between the two surveys (2.7% in 1996/7 to 4.6% in 1998/9). NRT abuse rates were well below those of other OTC abusable substances (e.g., diet pills and inhalants). CONCLUSIONS: Concerns over promotion of youth dependence to nicotine by offering the sale of NRT OTC to adults have not been realized and policymakers should consider reducing barriers to access these products. PMID- 16368661 TI - Efficacy of acupuncture for treating cocaine addiction: a review paper. AB - Acupuncture is being used in more than three hundred treatment facilities for treating substance abuse including cocaine addiction. Previous review papers could not evaluate the role of acupuncture for treating cocaine addiction because of lack of clinical trials at the time. Since then, several important studies were conducted in the field. This paper reviews existing clinical trials using acupuncture to treat cocaine addiction. The existing evidence fails to document the benefit of acupuncture in treating cocaine addiction as the sole treatment. Further efforts to document the efficacy of the NADA protocol to treat cocaine addiction as the sole treatment should be re-evaluated. There is a limited amount of evidence showing possible benefits of acupuncture when used as an adjunctive therapy to complement existing substance abuse treatment program. Future efforts should concentrate on systematic studies investigating the role of acupuncture as an adjunctive treatment for cocaine addiction. In addition, more formative research exploring the efficacy of different types and protocols of acupuncture treatments for cocaine addiction should be thoroughly investigated. PMID- 16368662 TI - On the nameless love and infinite sexualities: John Henry Mackay, Magnus Hirschfeld and the origins of the sexual emancipation movement. AB - Two prominent representatives of the sexual emancipation movement in Germany, John Henry Mackay (1864-1933) and Magnus Hirschfeld (1868-1935) launched significant attacks on sexual binarism and its combinatories. Although Mackay defended the nameless love against seminal Christian and subsequent secularised misconstructions of its nature, he was unable to overcome the fundamental scheme of binomic sexuality. Hirschfeld, however, resolved the theoretical issue through his doctrine of sexual intermediaries (Zwischenstufenlehre) which purports that without exception- all human beings are intersexual variants, i.e. unique composites of different proportions of masculinity and femininity. Since these proportions vary from one sexual layer of description to another in the same individual and can alter or be altered in time, it is sensu stricto not possible implies a radical deconstruction of not only binomic sexuality but its supplementation through a third sex. It offers a meta-theoretical framework for rethinking sexual difference beyond the fictional schemes and categorial closures of Western traditions of sexual identity. His assumption of potentially infinite sexualities anticipates some of the basic tenets forwarded by the philosophical and political agendas of queer studies. to postulate discrete sexual categories. Hirschfeld's doctrine. PMID- 16368663 TI - In search of a more perfect heteroarchy:Vermont, civil unions and the harm of "separate-but-equal". AB - This article focuses on the Vermont civil union solution to the state Supreme Court's mandate in Baker v. State (1999). Using non-subordination theory, the author argues that rather than being a legal victory for lesbians and gay men, the Vermont law integrally contributes to the maintenance of an imbalance of power between heterosexuals and lesbians and gays. The article analyzes the rhetorical strategy employed by lawmakers to respond to what they perceived and portrayed as a menace posed by same-sex marriage and demonstrates that lawmakers reinforced apprehensions surrounding lesbian and gay identity and asserted the familiar heterosexist narrative in an effort to quell the threat posed by Baker. The article concludes that the legislature's genuine motivation behind civil unions was validating their own and their constituents' misplaced fears regarding gay and lesbian identity, and pacifying those fears by denying equal marriage rights to gays and lesbians in an attempt to appear responsive to a perceived threat to heterosexual primacy. PMID- 16368664 TI - In from the outer generational differences in coming out and gay identity formation. AB - This paper explores generational differences among gay men in their experience of, and narratives relating the process of, coming out. The interview data for 32 gay men living in Melbourne, Australia, is drawn from the Community, Attachment, Structures and the epidemic (CASE) Study. Narratives of coming out for three generational groups: pre-AIDS, peri-AIDS, and post-AIDS, are examined in terms of the experiences of disclosure to family and friends. A shift from narratives organised around crisis and disjunction to those organised around consolidation is matched by differences in the degree to which peer networks are maintained post-coming out. PMID- 16368665 TI - Ex-gay rhetoric and the politics of sexuality: the Christian antigay/pro-family movement's "truth in love" ad campaign. AB - In 1998, a coalition of antigay, pro-family activist organizations published a set of full-page print advertisements in several nationally recognized newspapers. These ads promoted sexual (ex-gay) conversion therapy for homosexuals. I examine these advertisements as a contest over cultural symbols and values, and over the very definition of lesbian and gay identity. This discursive contest had the potential to impact activist politics greatly, but this impact was mitigated significantly by a similar set of ads produced in response by an opposing movement: the lesbian, gay, bisexual and transgender movement. The interactive dynamics between opposing movements impact the political field in which activists on each side pursue their goals. PMID- 16368666 TI - Cultural value orientations, internalized homophobia, and accommodation in romantic relationships. AB - In the present study, we examined the impact of cultural value orientations (i.e., the personally oriented value of individualism, and the socially oriented values of collectivism, familism, romanticism, and spiritualism) on accommodation (i.e., voice and loyalty, rather than exit and neglect, responses to partners' anger or criticism) in heterosexual and gay relationships; and we examined the impact of internalized homophobia (i.e., attitudes toward self, other, and disclosure) on accommodation specifically in gay relationships. A total of 262 heterosexuals (102 men and 162 women) and 857 gays (474 men and 383 women) participated in the present study. Consistent with hypotheses, among heterosexuals and gays, socially oriented values were significantly and positively related to accommodation (whereas the personally oriented value of individualism was unrelated to accommodation); and among gays in particular, internalized homophobia was significantly and negatively related to accommodation. Implications for the study of heterosexual and gay relationships are discussed. PMID- 16368667 TI - Consideration of future consequences and unprotected anal intercourse among men who have sex with men. AB - This study of men who have sex with men (MSM) examined whether tendencies to consider the future consequences of one's actions were associated with sexual behaviors that place oneself at risk for HIV infection. A total of 339 HIV negative MSM responded to the Consideration of Future Consequences Scale (CFC; Strathman et al., 1994) and to questions about their anal intercourse practices in the past year. In bivariate analyses, men with a stronger future orientation were less likely to engage in anal intercourse unprotected by a condom (p < .05). Multivariate analyses revealed that CFC accounted for significant variance in three of four measures of unprotected anal sex after statistically controlling for demographic covariates (education, income, ethnicity, age). CFC was a better predictor of sexual behavior and accounted for more unique variance than any of the demographic factors. Additional research is needed to confirm that CFC is an antecedent of behavior and to examine the feasibility and efficacy of focusing on CFC in HIV prevention interventions. PMID- 16368668 TI - "Risks both known and unknown"a qualitative method to assess the role of situation in HIV/STD risk and prevention. AB - The idea of situation has gained increased attention in HIV/STD prevention research and practice. In the context of prevention, situation does not simply connote setting or place but also incorporates meanings people attached to the physical setting and how the complex interrelation of setting, meaning, and behaviors influences decisions regarding sexual behaviors and prevention measures. Kenneth Burke's pentadic analysis provides a means to illuminate how situation influences decisions regarding sexual behaviors and risk taking. This manuscript describes the pentadic method, its application to situation and sexual risk behaviors, and its utility through content analysis of transcribed interviews (or texts) with men who patronize bathhouses, bathhouse staff members, and release forms from bathhouse establishments in three geographic areas. In this case, the analysis reveals that bathhouse patrons and policies perceive bathhouses as situations that privilege expedient sexual release rather than individuals and over how those releases occur. This predominant perspective impedes, but does not preclude, risk reduction. The authors provide recommendations for promoting condom use for anal sex in the bathhouse setting as well as for the utility of pentadic analysis in related research. PMID- 16368669 TI - Description of a domestic violence measure for Puerto Rican gay males. AB - A sample of 302 Puerto Rican gay males living in Puerto Rico and New York participated in this study with the objective of assessing the prevalence of domestic violence. A self-administered questionnaire was developed addressing issues of intergenerational violence, addictive behaviors, and domestic violence in three dimensions: emotional, physical, and sexual violence. The results concluded that close to half of the participants had experienced some sort of violence in their intimate relationships, have a history of being witness to domestic violence in their family of origin, and had identified in themselves and their families, addictive behaviors. Other variables measured are HIV and sexual coercion, drug and alcohol abuse, and levels of acculturation among participants living in the United States. The main objective of this work is to describe the development of the instrument used in the study. Based on the results of this study we describe the psychometric characteristics and content of the final questionnaire. Final recommendations are made for other researchers interested in doing domestic violence studies with Latino gay men and men who have sex with men. PMID- 16368672 TI - Older women's experiences of psychological violence in their marital relationships. AB - Violence within older couples is a reality, not a myth. In this study, qualitative interviews were conducted with 15 women ages 60 to 81 who had suffered marital psychological violence. Husbands' psychologically violent behaviors were grouped into 14 categories: control, denigration, deprivation, intimidation, threats, abdication of responsibility, manipulation, blame, harassment, negation of reality, indifference, making the wife feel guilty, sulking, and infantilization. Control behaviors were found to be the central category. Control dynamics increased at retirement, when children left home and when husbands experienced a decrease in health status. Implications for practice are also discussed. PMID- 16368673 TI - Differences in family functioning and health between older adult volunteers and non-volunteers. AB - The purpose of this investigation was to identify differences in family functioning and health between older adult volunteers and non-volunteers. Two findings emerged from the data: volunteers reported higher family functioning as well as better health compared to non-volunteers. Implications for social workers involved with volunteer programs and services are discussed in terms of volunteer recruitment and retention, and the related need to develop policies that permit more flexibility and choice with regard to volunteer responsibilities. PMID- 16368674 TI - HIV/AIDS fifty and older: a hidden and growing population. AB - Contrary to common belief, HIV and AIDS are prevalent in the older adult population. Currently, between ten to fifteen percent of HIV/AIDS new cases occur in individuals fifty and older (Chiao, Ries, & Sande, 1999). With the aging of the baby boomer generation, and the introduction of erectile dysfunction medications, such statistics are guaranteed to increase at alarming rates. This population also faces hazardous risk factors that increase its possibility of contracting the disease. Stereotypes and lack of information continue to limit older adults from receiving the medical, educational, and psychological attention they need to avoid and to combat the virus. This article discusses the rising trends in the fifty and older population as well as some of the factors that increase the prevalence of HIV/AIDS among this population. Prevention, assessment, and treatment practices are also outlined; if instituted, these interventions may help promote a significant reduction in the rate of HIV/AIDS infection in the older adult population. PMID- 16368675 TI - Negotiating relationship contexts in gerontological social work practice. AB - OBJECTIVES: The objective of this study was to demonstrate the role of social workers in addressing the complex relationship and problem solving contexts typical of practice in multi-service geriatric care settings. METHODS: We conducted a thematic analysis of the work of five social workers with five clients, their families, and the professional healthcare team, and extracted common themes across cases that illustrate the nature and timing of the interventions depending on the relationship context addressed. RESULTS: While there were problems and issues common across relationship contexts, intervention strategies that were essential for optimizing client care and wellbeing were specific to resolving relationship conflicts in one or more of the identified relationship contexts. PMID- 16368676 TI - Anticipated and experienced changes in activities after husbands retire. AB - As an element of anticipatory socialization, ability to predict future roles accurately may impact subsequent adaptation. Part of a larger study of retirement and marital quality, this longitudinal research examined husbands' and wives' (n = 61 couples) anticipations of change (more/less/same) in six individual and joint activities following husbands' retirement, and compared them with couples' reported experiences a year after husbands had retired. With the exception of household tasks, continuity in levels of activity from pre-to post-retirement was greater than couples had anticipated. Cross-classification of responses at baseline and Time 2 indicated only modest congruence between anticipated and experienced change in activities. Accuracy of anticipation was not related significantly to retirement satisfaction as hypothesized, but direction of retirement-satisfaction mean scores, especially among wives, suggest that future testing would be warranted. PMID- 16368677 TI - Vital Involvement Practice: strengths as more than tools for solving problems. AB - This article describes Vital Involvement Practice, a strength-based approach to clinical practice with elders, including those who are extremely frail. Using this approach, practitioners have been able to help elders increase later-life vitality and associated positive quality of life through: (1) systematic identification of individual strengths and assets (found both in the person and in the surrounding environment), and (2) consideration of these strengths alongside the individual and environmental deficits that are the subject of most geriatric practice. The approach utilizes original data-gathering tools (Occupational Profile; Life Strengths Interview Guide) and a stepwise, worksheet- structured consideration of these data in order to formulate action strategies for achieving client goals (Domain Scan; Domain Goals; Life Plan/Strategy). All elements of VIP emerged in pilot work with gerontological practitioners and their elder clients in such settings as: primary health care; government social service; subsidized senior housing; private clinical practice; community recreation. Limitations, implications, and promise are noted, with respect to practice and research. PMID- 16368679 TI - Mouse sphingosine kinase isoforms SPHK1a and SPHK1b differ in enzymatic traits including stability, localization, modification, and oligomerization. AB - Sphingosine kinases catalyze the production of the bioactive lipid molecule sphingosine 1-phosphate. Mice have two isoforms of sphingosine kinase type 1, SPHK1a and SPHK1b. In addition to the previously reported difference in their enzyme activities, we have found that these isoforms differ in several enzymatic characteristics. First, SPHK1b is unstable, whereas SPHK1a is highly stable. Degradation of SPHK1b occurs at the membrane and is inhibited by a proteasome inhibitor. Second, only SPHK1b exhibits abnormal mobility on SDS-PAGE, probably due to its SDS-resistant structure. Third, SPHK1a and SPHK1b are predominantly detected in the soluble and membrane fractions, respectively, when their degradation is inhibited. Fourth, only SPHK1b is modified with lipid, on its unique Cys residues (Cys-4 and Cys-5). Site-directed mutagenesis at these Cys residues resulted in increased sphingosine kinase activity, suggesting that the modification is inhibitory to the enzyme. Finally, SPHK1b tends to form homo oligomers, whereas most SPHK1a is presented as monomers. We have also determined that the lipid modification of SPHK1b is involved in its homo-oligomerization. Thus, although these two proteins differ only in a few N-terminal amino acid residues, their enzymatic traits are extremely different. PMID- 16368680 TI - Sensing of lysophospholipids by TRPC5 calcium channel. AB - TRPC calcium channels are emerging as a ubiquitous feature of vertebrate cells, but understanding of them is hampered by limited knowledge of the mechanisms of activation and identity of endogenous regulators. We have revealed that one of the TRPC channels, TRPC5, is strongly activated by common endogenous lysophospholipids including lysophosphatidylcholine (LPC) but, by contrast, not arachidonic acid. Although TRPC5 was stimulated by agonists at G-protein-coupled receptors, TRPC5 activation by LPC occurred downstream and independently of G protein signaling. The effect was not due to the generation of reactive oxygen species or because of a detergent effect of LPC. LPC activated TRPC5 when applied to excised membrane patches and thus has a relatively direct action on the channel structure, either because of a phospholipid binding site on the channel or because of sensitivity of the channel to perturbation of the bilayer by certain lipids. Activation showed dependence on side-chain length and the chemical head-group. The data revealed a previously unrecognized lysophospholipid sensing capability of TRPC5 that confers the property of a lipid ionotropic receptor. PMID- 16368681 TI - Domain architecture of protein-disulfide isomerase facilitates its dual role as an oxidase and an isomerase in Ero1p-mediated disulfide formation. AB - Native disulfide bond formation in eukaryotes is dependent on protein-disulfide isomerase (PDI) and its homologs, which contain varying combinations of catalytically active and inactive thioredoxin domains. However, the specific contribution of PDI to the formation of new disulfides versus reduction/rearrangement of non-native disulfides is poorly understood. We analyzed the role of individual PDI domains in disulfide bond formation in a reaction driven by their natural oxidant, Ero1p. We found that Ero1p oxidizes the isolated PDI catalytic thioredoxin domains, A and A' at the same rate. In contrast, we found that in the context of full-length PDI, there is an asymmetry in the rate of oxidation of the two active sites. This asymmetry is the result of a dual effect: an enhanced rate of oxidation of the second catalytic (A') domain and the substrate-mediated inhibition of oxidation of the first catalytic (A) domain. The specific order of thioredoxin domains in PDI is important in establishing the asymmetry in the rate of oxidation of the two active sites thus allowing A and A', two thioredoxin domains that are similar in sequence and structure, to serve opposing functional roles as a disulfide isomerase and disulfide oxidase, respectively. These findings reveal how native disulfide folding is accomplished in the endoplasmic reticulum and provide a context for understanding the proliferation of PDI homologs with combinatorial arrangements of thioredoxin domains. PMID- 16368682 TI - PAX2 activates WNT4 expression during mammalian kidney development. AB - The transcription factor PAX2 is expressed during normal kidney development and is thought to influence outgrowth and branching of the ureteric bud. Mice with homozygous null Pax2 mutations have developmental defects of the midbrain hindbrain region, optic nerve, and ear and are anephric. During nephrogenesis, PAX2 is also expressed by mesenchymal cells as they cluster and reorganize to form proximal elements of each nephron, but the function of PAX2 in these cells is unknown. In this study we hypothesized that PAX2 activates expression of WNT4, a secreted glycoprotein known to be critical for successful nephrogenesis. PAX2 protein was identified in distal portions of the "S-shaped" body, and the protein persists in the emerging proximal tubules of murine fetal kidney. PAX2 activated WNT4 promoter activity 5-fold in co-transfection assays with JTC12 cells derived from the proximal tubule. Inspection of the 5'-flanking sequence of the human WNT4 gene identified three novel PAX2 recognition motifs; each exhibited specific PAX2 protein binding in electromobility shift assays. Two motifs were contained within a completely duplicated 0.66-kb cassette. Transfection of JTC12 cells with a PAX2 expression vector was associated with a 7-fold increase in endogenous WNT4 mRNA. In contrast, Wnt4 mRNA was decreased by 60% in mesenchymal cell condensates of fetal kidney from mice with a heterozygous Pax2 mutation. We speculated that a key function of PAX2 is to activate WNT4 gene expression in metanephric mesenchymal cells as they differentiate to form elements of the renal tubules. PMID- 16368683 TI - High density lipoprotein endocytosis by scavenger receptor SR-BII is clathrin dependent and requires a carboxyl-terminal dileucine motif. AB - The high density lipoprotein (HDL) receptor Scavenger Receptor BII (SR-BII) is encoded by an alternatively spliced mRNA from the SR-BI gene and is expressed in various tissues. SR-BII protein differs from SR-BI only in the carboxyl-terminal cytoplasmic tail, which, as we showed previously, must contain a signal that confers predominant intracellular expression and rapid endocytosis of HDL. We have shown that SR-BII mediates HDL endocytosis through aclathrin-dependent, caveolae-independent pathway. Two candidate amino acid motifs were identified in the tail that could mediate association with clathrin-containing endocytic vesicles: a putative dileucine motif at position 492-493 and an overlapping tyrosine-based YXXZ motif starting at position 489. Although substitution of tyrosine at position 489 with alanine or histidine did not affect endocytosis, substitution L492A resulted in increased surface binding of HDL and reduced HDL particle endocytosis. Substitution L493A had a less dramatic effect. No other regions in the carboxyl-terminal tail appeared to contain motifs required for HDL endocytosis. Substitutions of leucine at position 492 with the hydrophobic amino acids valine or phenylalanine also reduced HDL endocytosis, stressing the importance of leucine at this position. Introducing the SR-BII YTPLL motif into the carboxyl-terminal cytoplasmic tail of SR-BI converted SR-BI into an endocytic receptor resembling SR-BII. These results demonstrated that SR-BII differs from SR-BI in subcellular localization and trafficking and suggest that the two isoforms differ in the manner in which they target ligands intracellularly. PMID- 16368684 TI - Euglena gracilis ribonucleotide reductase: the eukaryote class II enzyme and the possible antiquity of eukaryote B12 dependence. AB - Ribonucleotide reductases provide the building blocks for DNA synthesis. Three classes of enzymes are known, differing widely in amino acid sequence but with similar structural motives and allosteric regulation. Class I occurs in eukaryotes and aerobic prokaryotes, class II occurs in aerobic and anaerobic prokaryotes, and class III occurs in anaerobic prokaryotes. The eukaryote Euglena gracilis contains a class II enzyme (Gleason, F. K., and Hogenkamp, H. P. (1970) J. Biol. Chem. 245, 4894-4899) and, thus, forms an exception. Class II enzymes depend on vitamin B(12) for their activity. We purified the reductase from Euglena cells, determined partial peptide sequences, identified its cDNA, and purified the recombinant enzyme. Its amino acid sequence and general properties, including its allosteric behavior, were similar to the class II reductase from Lactobacillus leichmannii. Both enzymes belong to a distinct small group of reductases that unlike all other homodimeric reductases are monomeric. They compensate the loss of the second polypeptide of dimeric enzymes by a large insertion in the monomeric chain. Data base searching and sequence comparison revealed a homolog from the eukaryote Dictyostelium discoideum as the closest relative to the Euglena reductase, suggesting that the class II enzyme was present in a common, B(12)-dependent, eukaryote ancestor. PMID- 16368685 TI - Crystal structure of a bacterial type IB DNA topoisomerase reveals a preassembled active site in the absence of DNA. AB - Type IB DNA topoisomerases are found in all eukarya, two families of eukaryotic viruses (poxviruses and mimivirus), and many genera of bacteria. They alter DNA topology by cleaving and resealing one strand of duplex DNA via a covalent DNA-(3 phosphotyrosyl)-enzyme intermediate. Bacterial type IB enzymes were discovered recently and are described as poxvirus-like with respect to their small size, primary structures, and bipartite domain organization. Here we report the 1.75-A crystal structure of Deinococcus radiodurans topoisomerase IB (DraTopIB), a prototype of the bacterial clade. DraTopIB consists of an amino-terminal (N) beta sheet domain (amino acids 1-90) and a predominantly alpha-helical carboxyl terminal (C) domain (amino acids 91-346) that closely resemble the corresponding domains of vaccinia virus topoisomerase IB. The five amino acids of DraTopIB that comprise the catalytic pentad (Arg-137, Lys-174, Arg-239, Asn-280, and Tyr-289) are preassembled into the active site in the absence of DNA in a manner nearly identical to the pentad configuration in human topoisomerase I bound to DNA. This contrasts with the apoenzyme of vaccinia topoisomerase, in which three of the active site constituents are either displaced or disordered. The N and C domains of DraTopIB are splayed apart in an "open" conformation, in which the surface of the catalytic domain containing the active site is exposed for DNA binding. A comparison with the human topoisomerase I-DNA cocrystal structure suggests how viral and bacterial topoisomerase IB enzymes might bind DNA circumferentially via movement of the N domain into the major groove and clamping of a disordered loop of the C domain around the helix. PMID- 16368686 TI - Mechanism of the long range anti-silencing function of targeted histone acetyltransferases in yeast. AB - Transcriptionally silent chromatin in Saccharomyces cerevisiae is associated with histone hypoacetylation and is formed through the action of the Sir histone deacetylase complex. A histone acetyltransferase (HAT) targeted near silent chromatin can overcome silencing at a distance by increasing histone acetylation in a sizable region. However, how a tethered HAT acetylates distant nucleosomes has not been resolved. We demonstrate here that targeting the histone H3-specific HAT Gcn5p promotes acetylation of not only histone H3 but also histone H4 in a broad region. We also show that long range anti-silencing and histone acetylation by targeted HATs can be blocked by nucleosome-excluding sequences. These results are consistent with the contention that a tethered HAT promotes stepwise propagation of histone acetylation along the chromatin. Because histone hypoacetylation is key to the formation and maintenance of transcriptionally silent chromatin, it is believed that acetylation promoted by a targeted HAT disrupts silent chromatin thereby overcoming silencing. However, we show that the acetylated and transcriptionally active region created by a tethered HAT retains structural hallmarks of Sir-dependent silent chromatin and remains associated with Sir proteins indicating that tethered HATs overcome silencing without completely dismantling silent chromatin. PMID- 16368687 TI - Role of the serum response factor in regulating contractile apparatus gene expression and sarcomeric integrity in cardiomyocytes. AB - The serum response factor (SRF) is a transcriptional regulator required for mesodermal development, including heart formation and function. Previous studies have described the role of SRF in controlling expression of structural genes involved in conferring the myogenic phenotype. Recent studies by us and others have demonstrated embryonic lethal cardiovascular phenotypes in SRF-null animals, but have not directly addressed the mechanistic role of SRF in controlling broad regulatory programs in cardiac cells. In this study, we used a loss-of-function approach to delineate the role of SRF in cardiomyocyte gene expression and function. In SRF-null neonatal cardiomyocytes, we observed severe defects in the contractile apparatus, including Z-disc and stress fiber formation, as well as mislocalization and/or attenuation of sarcomeric proteins. Consistent with this, gene array and reverse transcription-PCR analyses showed down-regulation of genes encoding key cardiac transcriptional regulatory factors and proteins required for the maintenance of sarcomeric structure, function, and regulation. Chromatin immunoprecipitation analysis revealed that at least a subset of these proteins are likely regulated directly by SRF. The results presented here indicate that SRF is an essential coordinator of cardiomyocyte function due to its ability to regulate expression of numerous genes (some previously identified and at least 28 targets newly identified in this study) that are involved in multiple and disparate levels of sarcomeric function and assembly. PMID- 16368688 TI - Synthesis, processing, and composition of the virion-associated HTLV-1 reverse transcriptase. AB - It is not known whether the low infectivity and low virion-associated polymerase activity of human T-cell lymphotropic virus type-1 (HTLV-1) are due to the quantity or quality of the reverse transcriptase (RT), because the protein has not yet been fully characterized. We have developed anti-RT antibodies and constructed HTLV-1 expression plasmids that express truncated or hemagglutinin tagged Pol polyproteins to examine the maturation and composition of HTLV-1 RT. We detected virion-associated proteins corresponding to RT-integrase (IN) (pr98) and RT (p62) as well as smaller proteins containing the polymerase (p49) or RNase H domains. We have identified the amino acid sequences in the Pol polyprotein that are cleaved by HTLV-1 protease to yield RT and IN. We have also identified the cleavage sites within RT that give rise to the p49 polymerase fragment. Immunoprecipitation of an epitope-tagged p62 subunit coprecipitated p49, indicating that the HTLV-1 RT complex can exist as a p62/p49 heterodimer analogous to the RT of HIV-1 (p66/p51). PMID- 16368689 TI - Isoform specificity of ankyrin-B: a site in the divergent C-terminal domain is required for intramolecular association. AB - Ankyrins contain significant amino acid identity and are co-expressed in many cell types yet maintain unique functions in vivo. Recent studies have identified the highly divergent C-terminal domain in ankyrin-B as the key domain for driving ankyrin-B-specific functions in cardiomyocytes. Here we identify an intramolecular interaction between the C-terminal domain and the membrane-binding domain of ankyrin-B using pure proteins in solution and the yeast two-hybrid assay. Through extensive deletion and alanine-scanning mutagenesis we have mapped key residues for interaction in both domains. Amino acids (1597)EED(1599) located in the ankyrin-B C-terminal domain and amino acids Arg(37)/Arg(40) located in ANK repeat 1 are necessary for inter-domain interactions in yeast two-hybrid assays. Furthermore, conversion of amino acids EED(1597) to AAA(1597) leads to a loss of function in the localization of inositol 1,4,5-trisphosphate receptors in ankyrin B mutant cardiomyocytes. Physical properties of the ankyrin-B C-terminal domain determined by circular dichroism spectroscopy and hydrodynamic parameters reveal it is unstructured and highly extended in solution. Similar structural studies performed on full-length 220-kDa ankyrin-B harboring alanine substitutions, (1597)AAA(1599), reveal a more extended conformation compared with wild-type ankyrin-B. Taken together these results suggest a model of an extended and unstructured C-terminal domain folding back to bind and potentially regulate the membrane-binding domain of ankyrin-B. PMID- 16368690 TI - The Brl domain in Sec63p is required for assembly of functional endoplasmic reticulum translocons. AB - Protein translocation into the endoplasmic reticulum occurs at pore-forming structures known as translocons. In yeast, two different targeting pathways converge at a translocation pore formed by the Sec61 complex. The signal recognition particle-dependent pathway targets nascent precursors co translationally, whereas the Sec62p-dependent pathway targets polypeptides post translationally. In addition to the Sec61 complex, both pathways also require Sec63p, an integral membrane protein of the Hsp40 family, and Kar2p, a soluble Hsp70 located in the ER lumen. Using a series of mutant alleles, we demonstrate that a conserved Brl (Brr2-like) domain in the COOH-terminal cytosolic region of Sec63p is essential for function both in vivo and in vitro. We further demonstrate that this domain is required for assembly of two oligomeric complexes of 350 and 380 kDa, respectively. The larger of these corresponds to the heptameric "SEC complex" required for post-translational translocation. However, the 350-kDa complex represents a newly defined hexameric SEC' complex comprising Sec61p, Sss1p, Sbh1p, Sec63p, Sec71p, and Sec72p. Our data indicate that the SEC' complex is required for co-translational protein translocation across the yeast ER membrane. PMID- 16368691 TI - The Giardia duodenalis 14-3-3 protein is post-translationally modified by phosphorylation and polyglycylation of the C-terminal tail. AB - The flagellated protozoan Giardia duodenalis (syn. lamblia or intestinalis) has been chosen as a model parasite to further investigate the multifunctional 14-3 3s, a family of highly conserved eukaryotic proteins involved in many cellular processes, such as cell cycle, differentiation, apoptosis, and signal transduction pathways. We confirmed the presence of a single 14-3-3 homolog gene (g14-3-3) by an in silico screening of the complete genome of Giardia, and we demonstrated its constitutive transcription throughout the life stages of the parasite. We cloned and expressed the g14-3-3 in bacteria, and by protein-protein interaction assays we demonstrated that it is a functional 14-3-3. Using an anti peptide antibody raised against a unique 18-amino acid sequence at the N terminus, we observed variations both in the intracellular localization and in the molecular size of the native g14-3-3 during the conversion of Giardia from trophozoites to the cyst stage. An affinity chromatography, based on the 14-3-3 binding to the polypeptide difopein, was set to purify the native g14-3-3. By matrix-assisted laser desorption ionization mass spectroscopy analysis, we showed that polyglycylation, an unusual post-translational modification described only for tubulin, occurred at the extreme C terminus of the native g14-3-3 on Glu246, Glu247, or both and that the Thr214, located in the loop between helices 8 and 9, is phosphorylated. We propose that the addition of the polyglycine chain can promote the binding of g14-3-3 to alternative ligands and that the differential rate of polyglycylation/deglycylation during the encystation process can act as a novel mechanism to regulate the intracellular localization of g14-3-3. PMID- 16368692 TI - Snf1p-dependent Spt-Ada-Gcn5-acetyltransferase (SAGA) recruitment and chromatin remodeling activities on the HXT2 and HXT4 promoters. AB - We previously showed that the Spt-Ada-Gcn5-acetyltransferase (SAGA) complex is recruited to the activated HXT2 and HXT4 genes and plays a role in the association of TBP-associated factors. Using the HXT2 and HXT4 genes, we now present evidence for a functional link between Snf1p-dependent activation, recruitment of the SAGA complex, histone H3 removal, and H3 acetylation. Recruitment of the SAGA complex is dependent on the release of Ssn6p-Tup1p repression by Snf1p. In addition, we found that the Gcn5p subunit of the SAGA complex preferentially acetylates histone H3K18 on the HXT2 and HXT4 promoters and that Gcn5p activity is required for removal of histone H3 from the HXT4 promoter TATA region. In contrast, histone H3 removal from the HXT2 promoter does not require Gcn5p. In conclusion, although similar protein complexes are involved, induction of HXT2 and HXT4 displays important mechanistic differences. PMID- 16368695 TI - pH-dependent equilibrium between long lived near-UV intermediates of photoactive yellow protein. AB - The long lived intermediate (signaling state) of photoactive yellow protein (PYP(M)), which is formed in the photocycle, was characterized at various pHs. PYP(M) at neutral pH was in equilibrium between two spectroscopically distinct states. Absorption maxima of the acidic form (PYP(M)(acid)) and alkaline form (PYP(M)(alkali)) were located at 367 and 356 nm, respectively. Equilibrium was represented by the Henderson-Hasselbalch equation, in which apparent pK(a) was 6.4. Content of alpha- and/or beta-structure of PYP(M)(acid) was significantly greater than PYP(M)(alkali) as demonstrated by the molar ellipticity at 222 nm. In addition, changes in amide I and II modes of beta-structure in the difference Fourier transform infrared spectra for formation of PYP(M)(acid) was smaller than that of PYP(M)(alkali). The vibrational mode at 1747 cm(-1) of protonated Glu-46 was found as a small band for PYP(M)(acid) but not for PYP(M)(alkali), suggesting that Glu-46 remains partially protonated in PYP(M)(acid), whereas it is fully deprotonated in PYP(M)(alkali). Small angle x-ray scattering measurements demonstrated that the radius of gyration of PYP(M)(acid) was 15.7 Angstroms, whereas for PYP(M)(alkali) it was 16.2 Angstroms. These results indicate that PYP(M)(acid) assumes a more ordered and compact structure than PYP(M)(alkali). Binding of citrate shifts this equilibrium toward PYP(M)(alkali). UV-visible absorption spectra and difference infrared spectra of the long lived intermediate formed from E46Q mutant was consistent with those of PYP(M)(acid), indicating that the mutation shifts this equilibrium toward PYP(M)(acid). Alterations in the nature of PYP(M) by pH, citrate, and mutation of Glu-46 are consistently explained by the shift of the equilibrium between PYP(M)(acid) and PYP(M)(alkali). PMID- 16368693 TI - A new type of non-Ca2+-buffering Apo(a)-based fluorescent indicator for intraluminal Ca2+ in the endoplasmic reticulum. AB - Genetically encoded Ca2+ indicators are outstanding tools for the assessment of intracellular/organelle Ca2+ dynamics. Basically, most indicators contain the Ca2+-binding site of a (mutated) cytosolic protein that interacts with its natural (mutated) interaction partner upon binding of Ca2+. Consequently, a change in the structure of the sensor occurs that, in turn, alters the fluorescent properties of the sensor. Herein, we present a new type of genetically encoded Ca2+ indicator for the endoplasmic reticulum (ER) (apoK1-er (W. F. Graier, K. Osibow, R. Malli, and G. M. Kostner, patent application number 05450006.1 at the European patent office)) that is based on a single kringle domain from apolipoprotein(a), which is flanked by yellow and cyan fluorescent protein at the 3'- and 5'-ends, respectively. Notably, apoK1-er does not interact with Ca2+ itself but serves as a substrate for calreticulin, the main constitutive Ca2+-binding protein in the ER. ApoK1-er assembles with calreticulin and the protein disulfide isomerase ERp57 and undergoes a conformational shift in a Ca2+-dependent manner that allows fluorescence resonance energy transfer between the two fluorophores. This construct primarily offers three major advantages compared with the already existing probes: (i) it resolves perfectly the physiological range of the free Ca2+ concentration in the ER, (ii) expression of apoK1-er does not affect the Ca2+ buffering capacity of the ER, and (iii) apoK1-er is not inactivated by binding of constitutive interaction partners that prevent Ca2+-dependent conformational changes. These unique characteristics of apoK1-er make this sensor particularly attractive for studies on ER Ca2+ signaling and dynamics in which alteration of Ca2+ fluctuations by expression of any additional Ca2+ buffer essentially has to be avoided. PMID- 16368694 TI - Quantitative analysis of muscarinic acetylcholine receptor homo- and heterodimerization in live cells: regulation of receptor down-regulation by heterodimerization. AB - Although previous pharmacological and biochemical data support the notion that muscarinic acetylcholine receptors (mAChR) form homo- and heterodimers, the existence of mAChR oligomers in live cells is still a matter of controversy. Here we used bioluminescence resonance energy transfer to demonstrate that M(1), M(2), and M(3) mAChR can form constitutive homo- and heterodimers in living HEK 293 cells. Quantitative bioluminescence resonance energy transfer analysis has revealed that the cell receptor population in cells expressing a single subtype of M(1), M(2), or M(3) mAChR is predominantly composed of high affinity homodimers. Saturation curve analysis of cells expressing two receptor subtypes demonstrates the existence of high affinity M(1)/M(2), M(2)/M(3), and M(1)/M(3) mAChR heterodimers, although the relative affinity values were slightly lower than those for mAChR homodimers. Short term agonist treatment did not modify the oligomeric status of homo- and heterodimers. When expressed in JEG-3 cells, the M(2) receptor exhibits much higher susceptibility than the M(3) receptor to agonist-induced down-regulation. Coexpression of M(3) mAChR with increasing amounts of the M(2) subtype in JEG-3 cells resulted in an increased agonist induced down-regulation of M(3), suggesting a novel role of heterodimerization in the mechanism of mAChR long term regulation. PMID- 16368696 TI - A carboxyl-terminal determinant of the neuronal glutamate transporter, EAAC1, is required for platelet-derived growth factor-dependent trafficking. AB - The neuronal glutamate transporter, EAAC1 (excitatory amino acid carrier 1), undergoes rapid regulation after treatment with platelet-derived growth factor (PDGF) or phorbol ester in C6 glioma cells and neurons. A large intracellular pool of EAAC1 exists, from which transporters are redistributed to the cell surface in response to these signals. Here we show that PDGF had no effect on subcellular localization of the glial glutamate transporter, GLT-1, after transfection into C6 glioma cells. Chimeras consisting of domains from EAAC1 or GLT-1 were used to investigate structural motifs involved in PDGF-dependent redistribution of EAAC1. PDGF did not induce trafficking of an EAAC1 chimera containing the carboxyl-terminal domain of GLT-1; however, it did induce trafficking of a GLT-1 chimera containing the carboxyl-terminal domain of EAAC1. A truncated mutant of EAAC1 lacking 10 carboxyl-terminal amino acids was responsive to PDGF, whereas a mutant lacking 20 residues was not. Alanine substitution mutagenesis in this region revealed a short motif, (502)YVN(504), necessary for regulated trafficking. This motif was also involved in protein kinase C-dependent trafficking, as mutant transporters exhibited an attenuated response to phorbol ester. Interestingly, the presence of YVN in the homologous region of a nonresponsive chimera was not sufficient to confer regulated trafficking; however, the presence of a 12-amino acid motif starting at this Tyr residue was sufficient to confer responsiveness to PDGF. These studies identify a novel motif within the carboxyl terminus of EAAC1 which is required for regulated trafficking. The possibility that this motif targets EAAC1 to an intracellular, "regulated pool" is discussed. PMID- 16368698 TI - Evaluation of six fluoroquinolones for their capabilities in restricting the selection of resistant salmonellae. PMID- 16368697 TI - DNA reshaping by MukB. Right-handed knotting, left-handed supercoiling. AB - MukB is a bacterial SMC (structural maintenance of chromosome) protein required for faithful chromosome segregation in Escherichia coli. We report here that purified MukB introduces right-handed knots into DNA in the presence of type-2 topoisomerase, indicating that the protein promotes intramolecular DNA condensation. The pattern of generated knots suggests that MukB, similar to eukaryotic condensins, stabilizes large right-handed DNA loops. In contrast to eukaryotic condensins, however, the net supercoiling stabilized by MukB was negative. Furthermore, DNA reshaping by MukB did not require ATP. These data establish that bacterial condensins alter the shape of double-stranded DNA in vitro and lend support to the notions that the right-handed knotting is the most conserved biochemical property of condensins. Finally, we found that MukB can be eluted from a heparin column in two distinct forms, one of which is inert in DNA binding or reshaping. Furthermore, we find that the activity of MukB is reversibly attenuated during chromatographic separation. Thus, MukB has a unique set of topological properties, compared with other SMC proteins, and is likely to exist in two different conformations. PMID- 16368699 TI - Third-generation cephalosporin resistance among Gram-negative bacilli causing meningitis in neurosurgical patients: significant challenges in ensuring effective antibiotic therapy. AB - OBJECTIVES: The treatment of meningitis caused by Gram-negative bacilli in neurosurgical patients is a major challenge because of the complexity of these patients, the emergence of antibiotic resistance in many of the causative organisms and the restricted choice of antibiotics suitable for use, owing to a failure to achieve high enough concentrations in the CSF. We reviewed the incidence, aetiology, treatment and outcome of all patients with Gram-negative bacillary meningitis (GNBM) in our centre over a 7 year period. METHODS: Beaumont Hospital, Dublin is a 720 bed tertiary referral hospital and contains the national neurosurgical centre for the Republic of Ireland. The case notes and microbiological records of all patients with GNBM between 1998 and 2004 inclusive were reviewed retrospectively. Only patients with positive CSF culture and clinical features compatible with meningitis were included. RESULTS: Forty separate episodes of GNBM involving 34 different patients occurred during the study period. The most common causative organisms were Enterobacter spp. (35%), Escherichia coli (22.5%) and Pseudomonas aeruginosa (15%). Twenty-five per cent of isolates were resistant to third-generation cephalosporins. The median duration of treatment was 19.2 days and a combination of intravenous and intraventricular antibiotics was the most common treatment regimen used. Mortality directly related to GNBM was 2.5%. CONCLUSIONS: Although the mortality directly related to GNBM was low, the emergence of strains resistant to third generation cephalosporins represents a therapeutic challenge. Treatment with combined intravenous and intraventricular antibiotics is recommended for 2-3 weeks, but more studies are required to determine the optimal management of this difficult condition. PMID- 16368700 TI - Enhanced efficacy of pH-sensitive nystatin liposomes against Cryptococcus neoformans in murine model. AB - OBJECTIVES: To evaluate the efficacy of pH-sensitive liposomes of nystatin against Cryptococcus neoformans infection in a murine model. METHODS: In the present study, we investigated the antifungal activity of nystatin entrapped in pH-sensitive liposomes in a murine model. Mice infected with C. neoformans were treated with nystatin in neutral egg phosphatidylcholine (egg-PC) liposomes, as well as pH-sensitive nystatin liposomes. The anticryptococcal efficacy of liposomal formulations of nystatin was assessed by continued survival and colony forming units (cfu) in liver and brain of the treated mice. RESULTS: pH-sensitive liposomes of nystatin showed better efficacy compared with its free or egg-PC liposome form against C. neoformans infection in BALB/c mice. Mice treated with pH-sensitive nystatin liposomes showed 80% survival with less fungal burden in liver and brain of treated mice. However, there was only 40% survival in the group of animals treated with egg-PC liposome-intercalated nystatin, whereas its free form had poor efficacy with 20% survival. CONCLUSIONS: The enhanced anticryptococcal efficacy of the pH-sensitive nystatin liposomes can be attributed to the pH-dependent release of the drug in the low pH environment of lysosomes. The destabilization of the pH-sensitive liposomes in the acidic environment of macrophages results in the site-specific targeting of nystatin that improves its intracellular antifungal activity. PMID- 16368701 TI - Alterations of the penicillin-binding proteins and murM alleles of clinical Streptococcus pneumoniae isolates with high-level resistance to amoxicillin in Spain. AB - AIMS: The aim of this study was to analyse the nucleotide sequences of regions encoding the penicillin-binding domains of pbp1A, pbp2B and pbp2X genes and murM alleles from 14 selected amoxicillin-resistant Streptococcus pneumoniae isolates (MICs 8-16 mg/L) obtained in Spain. METHODS: PFGE and dideoxynucleotide chain termination sequencing were used. RESULTS: Analysis of PFGE profiles showed that the amoxicillin-resistant S. pneumoniae strains belonged to six different PFGE patterns including the Spain23F-1, Spain6B-2, Spain9V-3 and Spain(14)-5 international clones; however, 8 of the 14 strains belonged to the Spain9V-3 clone. These strains showed the typical changes in penicillin-binding proteins (PBPs) 1A and 2X and had 10 unique changes in the 590-641 region of PBP2B as described previously. Transformation experiments tried to incorporate the transpeptidase domain of PBP2B including the 590-641 region associated with amoxicillin-resistant pneumococci. Sequencing of the pbp2B genes revealed that part of the 3' region of the pbp2B sequence encoding a region of the domain (around amino acid 514-538 to the C terminus of PBP2B) did not recombine with the R6 pbp2B gene. The murM sequence analysis showed that 6, 6 and 2 amoxicillin resistant S. pneumoniae strains had murMA, murMB5 and murMB6 alleles, respectively. However, strains with murMB5 or murMB6 alleles showed a single mutation (N537D) in the 537-581 region of PBP2B, while strains with the murMA allele had 12 unique changes. CONCLUSIONS: Ten unique changes in the 590-641 region of PBP2B and no specific murM alleles were found in S. pneumoniae strains isolated in Spain with an amoxicillin MIC>or=8 mg/L (MICs from 6 to 12 mg/L by 1 mg/L step dilution). In addition, the presence of specific mutations in PBP2B seems to play a key role in the presence of different murM alleles in these amoxicillin-resistant pneumococcal strains. PMID- 16368702 TI - Haemodialysis and thermoregulation. PMID- 16368703 TI - Haemoperitoneum caused by bilateral renal cyst rupture in an ACKD peritoneal dialysis patient. PMID- 16368704 TI - The use of routinely collected computer data for research in primary care: opportunities and challenges. AB - INTRODUCTION: Routinely collected primary care data has underpinned research that has helped define primary care as a specialty. In the early years of the discipline, data were collected manually, but digital data collection now makes large volumes of data readily available. Primary care informatics is emerging as an academic discipline for the scientific study of how to harness these data. This paper reviews how data are stored in primary care computer systems; current use of large primary care research databases; and, the opportunities and challenges for using routinely collected primary care data in research. OPPORTUNITIES: (1) Growing volumes of routinely recorded data. (2) Improving data quality. (3) Technological progress enabling large datasets to be processed. (4) The potential to link clinical data in family practice with other data including genetic databases. (5) An established body of know-how within the international health informatics community. CHALLENGES: (1) Research methods for working with large primary care datasets are limited. (2) How to infer meaning from data. (3) Pace of change in medicine and technology. (4) Integrating systems where there is often no reliable unique identifier and between health (person-based records) and social care (care-based records-e.g. child protection). (5) Achieving appropriate levels of information security, confidentiality, and privacy. CONCLUSION: Routinely collected primary care computer data, aggregated into large databases, is used for audit, quality improvement, health service planning, epidemiological study and research. However, gaps exist in the literature about how to find relevant data, select appropriate research methods and ensure that the correct inferences are drawn. PMID- 16368705 TI - Rapamycin alleviates toxicity of different aggregate-prone proteins. AB - Many neurodegenerative diseases are caused by intracellular, aggregate-prone proteins, including polyglutamine-expanded huntingtin in Huntington's disease (HD) and mutant tau in fronto-temporal dementia/tauopathy. Previously, we showed that rapamycin, an autophagy inducer, enhances mutant huntingtin fragment clearance and attenuated toxicity. Here we show much wider applications for this approach. Rapamycin enhances the autophagic clearance of different proteins with long polyglutamines and a polyalanine-expanded protein, and reduces their toxicity. Rapamycin also reduces toxicity in Drosophila expressing wild-type or mutant forms of tau and these effects can be accounted for by reductions in insoluble tau. Thus, our studies suggest that the scope for rapamycin as a potential therapeutic in aggregate diseases may be much broader than HD or even polyglutamine diseases. PMID- 16368706 TI - Targeted disruption of glycerol kinase gene in mice: expression analysis in liver shows alterations in network partners related to glycerol kinase activity. AB - Glycerol kinase deficiency (GKD) is an X-linked inborn error of metabolism with metabolic and neurological crises. Liver shows the highest level of glycerol kinase (GK) activity in humans and mice. Absence of genotype-phenotype correlations in patients with GKD indicates the involvement of modifier genes, including other network partners. To understand the molecular pathogenesis of GKD, we performed microarray analysis on liver mRNA from neonatal glycerol kinase (Gyk) knockout (KO) and wild-type (WT) mice. Unsupervised learning revealed that the overall gene expression profile of the KO mice was different from that of WT. Real-time PCR confirmed the differences for selected genes. Functional gene enrichment analysis was used to find 56 increased and 37 decreased gene functional categories. PathwayAssist analysis identified changes in gene expression levels of genes involved in organic acid metabolism indicating that GK was part of the same metabolic network which correlates well with the patients with GKD having metabolic acidemia during their episodic crises. Network component analysis (NCA) showed that transcription factors sterol regulatory element-binding protein (SREBP)-1c, carbohydrate response element-binding protein (ChREBP), hepatocyte nuclear factor-4 alpha (HNF-4alpha) and peroxisome proliferative-activated receptor-alpha (PPARalpha) had increased activity in the Gyk KO mice compared with WT mice, whereas SREBP-2 was less active in the Gyk KO mice. These studies show that Gyk deletion causes alterations in expression of genes in several regulatory networks and is the first time NCA has been used to expand on microarray data from a mouse KO model of a human disease. PMID- 16368707 TI - A human imprinting centre demonstrates conserved acquisition but diverged maintenance of imprinting in a mouse model for Angelman syndrome imprinting defects. AB - Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are caused by the loss of imprinted gene expression from chromosome 15q11-q13. Imprinted gene expression in the region is regulated by a bipartite imprinting centre (IC), comprising the PWS IC and the AS-IC. The PWS-IC is a positive regulatory element required for bidirectional activation of a number of paternally expressed genes. The function of the AS-IC appears to be to suppress PWS-IC function on the maternal chromosome through a methylation imprint acquired during female gametogenesis. Here we have placed the entire mouse locus under the control of a human PWS-IC by targeted replacement of the mouse PWS-IC with the equivalent human region. Paternal inheritance of the human PWS-IC demonstrates for the first time that a positive regulatory element in the PWS-IC has diverged. These mice show postnatal lethality and growth deficiency, phenotypes not previously attributed directly to the affected genes. Following maternal inheritance, the human PWS-IC is able to acquire a methylation imprint in mouse oocytes, suggesting that acquisition of the methylation imprint is conserved. However, the imprint is lost in somatic cells, showing that maintenance has diverged. This maternal imprinting defect results in expression of maternal Ube3a-as and repression of Ube3a in cis, providing evidence that Ube3a is regulated by its antisense and creating the first reported mouse model for AS imprinting defects. PMID- 16368708 TI - A PYY Q62P variant linked to human obesity. AB - Peptide YY (PYY) has been implicated in the control of food intake through functional studies in rodents and humans. To investigate whether genetic alterations within this gene result in abnormal weight in humans, we sequenced its coding exons and splice sites in a large cohort of extremely obese [n = 379; average body mass index (BMI), 49.0 kg/m2] and lean (n = 378; average BMI, 19.5 kg/m2) individuals. In total, three rare non-synonymous variants were identified, only one of which, PYY Q62P, exhibited familial segregation with body mass. Through serendipity, previous studies based on cell culture revealed this precise variant to have altered receptor-binding selectivity in vitro. We further show, using mouse peptide injection experiments, that while the wild-type PYY peptide reduces food intake, the mutant PYY 62P had an insignificant effect in reducing food intake in vivo. Taken together, these results are the first to support that rare sequence variants within PYY can influence human susceptibility to obesity. PMID- 16368709 TI - Mitochondrial and nuclear DNA defects in Saccharomyces cerevisiae with mutations in DNA polymerase gamma associated with progressive external ophthalmoplegia. AB - A number of nuclear mutations have been identified in a variety of mitochondrial diseases including progressive external ophthalmoplegia (PEO), Alpers syndrome and other neuromuscular and oxidative phosphorylation defects. More than 50 mutations have been identified in POLG, which encodes the human mitochondrial DNA (mtDNA) polymerase gamma, PEO and Alpers patients. To rapidly characterize the effects of these mutations, we have developed a versatile system that enables the consequences of homologous mutations, introduced in situ into the yeast mtDNA polymerase gene MIP1, to be evaluated in vivo in haploid and diploid cells. Overall, distinct phenotypes for expression of each of the mip1-PEO mutations were observed, including respiration-defective cells with decreased viability, dominant-negative mutant polymerases, elevated levels of mitochondrial and nuclear DNA damage and chromosomal mutations. Mutations in the polymerase domain caused the most severe phenotype accompanied by loss of mtDNA and cell viability, whereas the mutation in the exonuclease domain showed mild dominance with loss of mtDNA. Interestingly, the linker region mutation caused elevated mitochondrial and nuclear DNA damage. The cellular processes contributing to these observations in the mutant yeast cells are potentially relevant to understanding the pathologies observed in human mitochondrial disease patients. PMID- 16368710 TI - Tagging-SNP haplotype analysis of the secretory PLA2IIa gene PLA2G2A shows strong association with serum levels of sPLA2IIa: results from the UDACS study. AB - Recent prospective analysis identified secretory phospholipase A(2)-IIa (sPLA(2)IIa) as a coronary artery disease (CAD) risk predictor. This study aimed to examine the relationship between serum levels of sPLA(2)IIa and variation in the sPLA(2)IIa gene (PLA2G2A) in a cohort of patients with Type II diabetes (T2D) mellitus. Six tagging single nucleotide polymorphisms (tSNPs) accounting for > 92% of the genetic variability in PLA2G2A were identified and distinguished six common haplotypes (frequencies > 5%). In the 523 Caucasian T2D patients, levels of sPLA(2)IIa, independent of CRP, were negatively correlated with total antioxidant status (P = 0.003) and high-density lipoprotein cholesterol (P = 0.006) in men and correlated with CAD status in women (P = 0.002) (Odds ratio of top two tertiles versus bottom = 2.50) [95% CI (1.13-5.53) P = 0.024]. Overall, tSNP haplotypes showed a highly significant association with sPLA(2)IIa levels (P < 0.0001), explaining 6.3% of the variance. The most common haplotype (frequency 14.2%) was associated with 53% higher sPLA(2)IIa levels [3.25 ng/ml (+/- 0.14)] compared with the combined other haplotypes [2.13 ng/ml (+/- 0.09), P < 0.00001]. Five of the six tSNPs were associated with significant effects on sPLA(2)IIa levels but the raising haplotype could not be distinguished by a single tSNP and none are likely to be functional. These data confirm the relationship between elevated sPLA(2)IIa levels and CAD risk reported in both cases: control and prospective analyses. The strong impact of PLA2G2A haplotypic variation on sPLA(2)IIa levels will help clarify the causality of this association. PMID- 16368711 TI - A mutation in the small heat-shock protein HSPB1 leading to distal hereditary motor neuronopathy disrupts neurofilament assembly and the axonal transport of specific cellular cargoes. AB - Distal hereditary motor neuronopathies (dHMNs) are a clinically and genetically heterogeneous group of disorders in which motor neurons selectively undergo age dependant degeneration. Mutations in the small heat-shock protein HSPB1 (HSP27) are responsible for one form of dHMN. In this study, we have analysed the effect of expressing a form of mutant HSPB1 in primary neuronal cells in culture. Mutant (P182L) but not wild-type HSPB1 led to the formation of insoluble intracellular aggregates and to the sequestration in the cytoplasm of selective cellular components, including neurofilament middle chain subunit (NF-M) and p150 dynactin. These findings suggest a possible pathogenic mechanism for HSPB1 whereby the mutation may lead to preferential motor neuron loss by disrupting selective components essential for axonal structure and transport. PMID- 16368713 TI - Second primary cancers in patients with stage III non-small cell lung cancer successfully treated with chemo-radiotherapy. AB - BACKGROUND: Patients successfully treated for non-small cell lung cancer (NSCLC) remain at risk for developing second primary cancer (SPC). The purpose of the current study is to assess the incidence of SPC and the impact of smoking status on the SPC in long-term survivors with stage III NSCLC after chemo-radiotherapy. METHODS: Using the database from the Japan National Hospital Lung Cancer Study Group between 1985 and 1995, information was obtained on 62 patients who were more than 3 years disease-free survivors. Details of clinical information and most smoking history were available from the questionnaire. RESULTS: Nine of the 62 patients developed SPC 3.9-12.2 years (median, 6.2 years) after the initiation of the treatment. The site of SPC was 2 lung, 1 esophagus, 2 stomach, 1 colon, 1 breast, 1 skin and 1 leukemia. Among these nine, three cancers occurred inside the radiation field. The relative risk of any SPC was 2.8 [95% confidence interval (CI) 1.3-5.3]. The risk changed with the passage of time and it increased significantly (5.2 times at or beyond 7 years) after the treatment. In univariate analysis, the patients who were male, had more cumulative smoking and continued smoking, had an increased risk of SPC [relative risk (RR) 2.7, CI 1.1 5.3; RR 3.0, CI 1.2-6.2; RR 5.2, CI 1.6-11.7, respectively]. In multivariate analysis, factors including smoking status and histological type had no effect on the development of a SPC. CONCLUSION: The patients with stage III NSCLC successfully treated with chemo-radiotherapy were at risk for developing SPC and this risk increased with time. PMID- 16368712 TI - Palmitoyl-protein thioesterase-1 deficiency mediates the activation of the unfolded protein response and neuronal apoptosis in INCL. AB - Numerous proteins undergo modification by palmitic acid (S-acylation) for their biological functions including signal transduction, vesicular transport and maintenance of cellular architecture. Although palmitoylation is an essential modification, these proteins must also undergo depalmitoylation for their degradation by lysosomal proteases. Palmitoyl-protein thioesterase-1 (PPT1), a lysosomal enzyme, cleaves thioester linkages in S-acylated proteins and removes palmitate residues facilitating the degradation of these proteins. Thus, inactivating mutations in the PPT1 gene cause infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative storage disorder of childhood. Although rapidly progressing brain atrophy is the most dramatic pathological manifestation of INCL, the molecular mechanism(s) remains unclear. Using PPT1-knockout (PPT1-KO) mice that mimic human INCL, we report here that the endoplasmic reticulum (ER) in the brain cells of these mice is structurally abnormal. Further, we demonstrate that the level of growth-associated protein-43 (GAP-43), a palmitoylated neuronal protein, is elevated in the brains of PPT1-KO mice. Moreover, forced expression of GAP-43 in PPT1-deficient cells results in the abnormal accumulation of this protein in the ER. Consistent with these results, we found evidence for the activation of unfolded protein response (UPR) marked by elevated levels of phosphorylated translation initiation factor, eIF2alpha, increased expression of chaperone proteins such as glucose-regulated protein-78 and activation of caspase-12, a cysteine proteinase in the ER, mediating caspase-3 activation and apoptosis. Our results, for the first time, link PPT1 deficiency with the activation of UPR, apoptosis and neurodegeneration in INCL and identify potential targets for therapeutic intervention in this uniformly fatal disease. PMID- 16368714 TI - Exercise increases MEF2- and GEF DNA-binding activity in human skeletal muscle. AB - Overexpression of GLUT4 exclusively in skeletal muscle enhances insulin action and improves glucose homeostasis. Transgenic studies have discovered two regions on the GLUT4 promoter conserved across several species that are required for normal GLUT4 expression in skeletal muscle. These regions contain binding motifs for the myocyte enhancer factor 2 (MEF2) family and GLUT4 enhancer factor (GEF). A single bout of exercise increases both GLUT4 transcription and mRNA abundance; however, the molecular mechanisms mediating this response remain largely unexplored. Thus, the aim of this study was to determine whether a single, acute bout of exercise increased the DNA-binding activities of MEF2 and GEF in human skeletal muscle. Seven subjects performed 60 min of cycling at approximately 70% of VO2peak. After exercise, the DNA-binding activities of both the MEF2A/D heterodimer and GEF were increased (P<0.05). There was no change in nuclear MEF2D or GEF abundance after exercise, but nuclear MEF2A abundance was increased (P<0.05). These data demonstrate that exercise increases MEF2 and GEF DNA binding and imply that these transcription factors could be potential targets for modulating GLUT4 expression in human skeletal muscle. PMID- 16368715 TI - Insulin stimulates L-carnitine accumulation in human skeletal muscle. AB - Increasing skeletal muscle carnitine content may alleviate the decline in muscle fat oxidation seen during intense exercise. Studies to date, however, have failed to increase muscle carnitine content, in healthy humans, by dietary or intravenous L-carnitine administration. We hypothesized that insulin could augment Na+-dependent skeletal muscle carnitine transport. On two randomized visits, eight healthy men underwent 5 h of intravenous L-carnitine infusion with serum insulin maintained at fasting (7.4+/-0.4 mIU*l(-1)) or physiologically high (149.2+/-6.9 mIU*l(-1)) concentrations. The combination of hypercarnitinemia (approximately 500 micromol*l(-1)) and hyperinsulinemia increased muscle total carnitine (TC) content from 22.0 +/- 0.9 to 24.7 +/- 1.4 mmol*(kg dm)(-1) (P<0.05) and was associated with a 2.3 +/- 0.3-fold increase in carnitine transporter protein (OCTN2) mRNA expression (P<0.05). Hypercarnitinemia in the presence of a fasting insulin concentration had no effect on either of these parameters. This study demonstrates that insulin can acutely increase muscle TC content in humans during hypercarnitinemia, which is associated with an increase in OCTN2 transcription. These novel findings may be of importance to the regulation of muscle fat oxidation during exercise, particularly in obesity and type 2 diabetes where it is known to be impaired. PMID- 16368716 TI - Pigment epithelium-derived factor (PEDF) is an endogenous antiinflammatory factor. AB - Pigment epithelium-derived factor (PEDF) is a potent angiogenic inhibitor. Reduced PEDF levels are associated with diabetic retinopathy. However, the mechanism for the protective effects of PEDF against diabetic retinopathy (DR) is presently unclear. As inflammation plays a role in DR, the present study determined the effect of PEDF on inflammation. Western blot analysis and ELISA demonstrated that retinal and plasma PEDF levels were drastically decreased in rats with endotoxin-induced uveitis (EIU), which suggests that PEDF is a negative acute-phase protein. Intravitreal injection of PEDF significantly reduced vascular hyper-permeability in rat models of diabetes and oxygen-induced retinopathy, correlating with the decreased levels of retinal inflammatory factors, including VEGF, VEGF receptor-2, MCP-1, TNF-alpha, and ICAM-1. In cultured retinal capillary endothelial cells, PEDF significantly decreased TNF alpha and ICAM-1 expression under hypoxia. Moreover, down-regulation of PEDF expression by siRNA resulted in significantly increases of VEGF and TNF-alpha secretion in retinal Muller cells. These findings suggest that PEDF is a novel endogenous anti-inflammatory factor in the eye. The decrease of ocular PEDF levels may contribute to inflammation and vascular leakage in DR. PMID- 16368717 TI - Role of nuclear receptor signaling in platelets: antithrombotic effects of PPARbeta. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors that heterodimerize with the retinoid X receptor and then modulate at the transcriptional level the function of many target genes. Three PPARs are known: alpha, beta (sometimes called delta), and gamma. The better studied are PPARalpha and PPARgamma, which are activated by fibrates and thiazolidinediones/glitazones, respectively. It is now believed that activation of the PPARs could be associated with the prevention of heart attack and stroke in humans. Here we report, for the first time, that human platelets contain PPARbeta and that its selective activation inhibits platelet aggregation. PPARbeta is a putative receptor for prostacyclin. Prostacyclin is an important antithrombotic hormone that synergizes with nitric oxide to inhibit platelet aggregation. In the current study, we show that PPARbeta ligands similarly synergize with nitric oxide to inhibit platelet aggregation. These observations challenge our view of a nuclear receptor because PPARbeta is present and active in nonnucleated platelets. Furthermore, these data suggest that some of the antithrombotic actions of prostacyclin may be mediated via activation of PPARs. Thus, our results identify PPARbeta as a novel antiplatelet target that may mediate some of the effects of prostacyclin in blood. PMID- 16368718 TI - Endothelial activation and induction of monocyte adhesion by nontransferrin-bound iron present in human sera. AB - Nontransferrin-bound iron (NTBI) has been detected in iron overload diseases. This form of iron may exert pro-oxidant effects and modulate cellular function and inflammatory response. The present study has aimed to investigate the effects of serum NTBI on monocyte adherence to endothelium. Measured by a recently developed high-throughput fluorescence-based assay, serum NTBI was found to be higher in both homozygotes of HFE C282Y mutation of hereditary hemochromatosis (7.9+/-0.6 microM, n=9, P<0.001) and heterozygotes (4.0+/-0.5 microM, n=8, P<0.001), compared with controls (1.6+/-0.2 microM, n=21). The effects of these sera on monocyte adhesion and endothelial activation were examined. Adhesion of normal human monocytes to C282Y homozygote- and heterozygote-serum-treated human umbilical vein endothelial cells was higher (25.0+/-0.9 and 22.1+/-0.7%, respectively) compared with controls (17.6+/-0.5%, both P<0.001). For the three groups combined, the expression of adhesion molecules, ICAM-1, VCAM-1, and E selectin, was positively correlated to NTBI levels but not to the inflammatory marker C-reactive protein. Furthermore, accumulation of intracellular labile iron and oxidative radicals within the cells due to NTBI was evidenced. Finally, counteraction of NTBI-induced endothelial activation was observed using iron chelators. These findings therefore identify a physiological function of NTBI in monocyte-endothelial interactions that may also contribute to the development of atherosclerosis and neurodegenerative diseases. PMID- 16368719 TI - Beta-adrenergic receptor trafficking by exercise in rat adipocytes: roles of G protein-coupled receptor kinase-2, beta-arrestin-2, and the ubiquitin-proteasome pathway. AB - The effect of exercise on beta-adrenergic receptor (beta-AR) trafficking was investigated in rat adipocytes. The binding sites of a hydrophilic ligand, [(3)H]CGP12177, increased immediately (0 h) and at 3 h after exercise (3 h) but decreased at 24 h after exercise (24 h). The data of immunoblotting revealed that the alterations in the binding sites mainly paralleled the alterations in the beta2-AR proteins in membrane fractions. The protein expressions of both G protein-coupled receptor kinase (GRK)-2 and beta-arrestin-2 were reduced, with a decline in beta2-AR ubiquitination at 0 h and 3 h. The protein expressions of beta2-AR, GRK-2, beta-arrestin-2, the beta2-AR/beta-arrestin-2 complex, and beta2 AR ubiquitination returned to their respective control levels at 24 h, whereas the beta2-AR mRNA level was reduced. Administration of either lactacystin or propranolol did not alter GRK-2 and beta2-AR protein expressions after exercise. Thus, the mechanism underlying the increased density of beta2-AR up to at least 3 h may involve alterations in a multistep event involving the coordinate interaction among proteins mediating beta2-AR trafficking, in which both the receptor-agonist interactions and ubiquitin-proteasome pathway have a key role. However, the decreased protein expression of beta2-AR at 24 h might be due to some change occurring at the translational levels. PMID- 16368720 TI - Shifting the optimal pH of activity for a laccase from the fungus Trametes versicolor by structure-based mutagenesis. AB - Laccases are oxidizing enzymes of interest because of their potential environmental and industrial applications. We performed site-directed mutagenesis of a laccase produced by Trametes versicolor in order to improve its catalytic properties. Considering a strong interaction of the Asp residue in position 206 with the substrate xylidine, we replaced it with Glu, Ala or Asn, expressed the mutant enzymes in the yeast Yarrowia lipolytica and assayed the transformation of phenolic and non-phenolic substrates. The transformation rates remain within the same range whatever the mutation of the laccase and the type of substrate: at most a 3-fold factor increase was obtained for k(cat) between the wild-type and the most efficient mutant Asp206Ala with 2,2'-azinobis(3-ethylbenzthiazoline-6 sulfonic) acid as a substrate. Nevertheless, the Asn mutation led to a significant shift of the pH (DeltapH = 1.4) for optimal activity against 2,6 dimethoxyphenol. This study also provides a new insight into the binding of the reducing substrate into the active T1 site and induced modifications in catalytic properties of the enzyme. PMID- 16368721 TI - An improved method for an efficient and easily accessible eukaryotic ribosome display technology. AB - Ribosome display is a powerful in vitro technology for the selection and directed evolution of proteins. However, this technology has so far been perceived as being technically challenging owing to comparatively difficult protocols and the absence of tailored commercial reagents, particularly when using prokaryotic cell free expression systems. Eukaryotic ribosome display is potentially a more accessible alternative because of the availability of suitable commercial reagents, yet despite published protocols, this method has been less widely used. For eukaryotic ribosome display, a novel mechanism of mRNA recovery compared with that of the well-proven prokaryotic method has been proposed. We have examined the eukaryotic ribosome display process with the aims of investigating the proposed mechanism of sequence recovery and of identifying aspects of the protocol that may have lead to poor performance and therefore so far limited its use. We demonstrate that the proposed novel method is in fact mechanistically comparable to the prokaryotic method and we provide a step-by-step protocol for eukaryotic ribosome display that is 20-fold more efficient than current published methods. Our findings should increase the ease of operating ribosome display technology, making it more accessible to the scientific community. PMID- 16368722 TI - The Evra (ethinyl estradiol/norelgestromin) contraceptive patch: estrogen exposure concerns. PMID- 16368723 TI - Commentary: benefits of influenza vaccine in US elderly--new studies raise questions. PMID- 16368724 TI - Functional status is a confounder of the association of influenza vaccine and risk of all cause mortality in seniors. AB - BACKGROUND: Functional status limitations may be associated with both an increased risk of death and a decreased likelihood of influenza vaccination, and so may confound the association of influenza vaccination and risk of all cause mortality in seniors. METHODS: We conducted a nested case-control study of persons >or=65 years of age that included 252 cases who died during an influenza season and 576 age-matched controls. We identified functional limitations by medical record review, and compared the effect of adjustment for those factors with that of adjustment for disease covariates defined by diagnosis codes, using methods reported by previous influenza vaccine effectiveness studies, on the association of influenza vaccination and risk of death. RESULTS: Functional limitations, such as requiring assistance for bathing, were highly prevalent in cases, even in the subgroup defined as free of comorbidity by diagnosis code criteria, and were associated with a decreased likelihood of vaccination among controls. Adjustment for functional limitations resulted in an estimate of the relative risk of death in vaccinated persons compared with unvaccinated persons that was closer to the null [odds ratio (OR), 0.71; 95% confidence interval (95% CI), 0.47-1.06] than the unadjusted estimate (OR, 0.59; 95% CI, 0.41-0.83). In contrast, adjustment for diagnosis code covariates moved the estimate further from the null (OR, 0.45; 95% CI, 0.30-0.68). CONCLUSIONS: Functional limitations appear to be important confounders of the association of vaccination and risk of death, while adjustment for diagnosis code covariates did not control for a healthy vaccinee bias. Further research is needed on methods to reduce the influence of bias in observational studies of influenza vaccine effectiveness. PMID- 16368725 TI - Evidence of bias in estimates of influenza vaccine effectiveness in seniors. AB - BACKGROUND: Numerous observational studies have reported that seniors who receive influenza vaccine are at substantially lower risk of death and hospitalization during the influenza season than unvaccinated seniors. These estimates could be influenced by differences in underlying health status between the vaccinated and unvaccinated groups. Since a protective effect of vaccination should be specific to influenza season, evaluation of non-influenza periods could indicate the possible contribution of bias to the estimates observed during influenza season. METHODS: We evaluated a cohort of 72,527 persons 65 years of age and older followed during an 8 year period and assessed the risk of death from any cause, or hospitalization for pneumonia or influenza, in relation to influenza vaccination, in periods before, during, and after influenza seasons. Secondary models adjusted for covariates defined primarily by diagnosis codes assigned to medical encounters. RESULTS: The relative risk of death for vaccinated persons compared with unvaccinated persons was 0.39 [95% confidence interval (95% CI), 0.33-0.47] before influenza season, 0.56 (0.52-0.61) during influenza season, and 0.74 (0.67-0.80) after influenza season. The relative risk of pneumonia hospitalization was 0.72 (0.59-0.89) before, 0.82 (0.75-0.89) during, and 0.95 (0.85-1.07) after influenza season. Adjustment for diagnosis code variables resulted in estimates that were further from the null, in all time periods. CONCLUSIONS: The reductions in risk before influenza season indicate preferential receipt of vaccine by relatively healthy seniors. Adjustment for diagnosis code variables did not control for this bias. In this study, the magnitude of the bias demonstrated by the associations before the influenza season was sufficient to account entirely for the associations observed during influenza season. PMID- 16368726 TI - The use of general practice consultation databases in rheumatology research. PMID- 16368727 TI - Infliximab treatment in a patient with rheumatoid arthritis on haemodialysis. PMID- 16368728 TI - Cost-effective detection of non-antidouble-stranded DNA antinuclear antibody specificities in daily clinical practice. AB - OBJECTIVES: To compare the utility of indirect immunofluorescence for the detection of antinuclear antibodies (ANA-IIF) and a fully automated test (ELiA Symphony) that detects antibodies against a mixture of nuclear and cytoplasmic antigens (ENA), to select sera that should be tested for non-antidouble-stranded DNA (dsDNA) antinuclear antibodies in a relatively expensive automated line immunoassay (INNO-LIA ANA update, Lineblot). METHODS: All 328 sera sent to the laboratory for ANA or anti-ENA tests, over a 4 month period were evaluated in all three assays. Results were related to signs and symptoms of systemic autoimmune disease (AID) that patients had before or at the time of blood sampling. RESULTS: Overall, 72 (22%) sera were Lineblot positive. Of 198 patients without clinical manifestations of AID, 7% were Lineblot positive. Limiting Lineblot to sera positive in either ANA-IIF or Symphony tests failed to detect 26 (ANA-IIF) and 22 (Symphony) Lineblot-reactive sera, with 15 sera being negative in both assays. From a clinical point of view, failure to detect these reactivities was not important in most cases. CONCLUSIONS: Restriction of performance of Lineblot to patients with at least one criterion for AID is an ideal and cost-effective strategy. In ignorance of clinical signs and symptoms, screening of sera by ANA IIF or Symphony strongly reduces the costs of anti-ENA detection, with minimal loss in diagnostic capacity. Based on small differences, including the fact that anti-dsDNA antibodies give a positive ANA-IIF, we prefer screening with ANA-IIF over Symphony. PMID- 16368729 TI - The epidemiology of Wegener's granulomatosis and microscopic polyangiitis in a Southern Hemisphere region. AB - OBJECTIVE: To determine the prevalence of Wegener's granulomatosis (WG) and microscopic polyangiitis (MPA) in the province of Canterbury, New Zealand. METHOD: Three hospital clinical databases and the immunology laboratory database were searched and case notes reviewed for patients fulfilling either the 1990 American College of Rheumatology (ACR) criteria for WG or a modification of those criteria that allowed for antineutrophil cytoplasmic antibody (ANCA) positivity in the absence of granulomatous vasculitis. MPA was defined by the Chapel Hill consensus definition; however, in the absence of histological evidence of pauci immune glomerulonephritis, ANCA positivity in association with evidence of active glomerular disease was included as a criterion. The point prevalence at 31 December 2003 and the 5-yr period prevalence for the interval 1 January 1999 to 31 December 2003 were calculated. RESULTS: Seventy-three patients with WG and 28 patients with MPA fulfilled the inclusion criteria. A 5-yr period prevalence of 152 WG cases/million [95% confidence interval (CI) 117-186] and 58 MPA cases/million (95% CI 37-80) was calculated using 2001 census data as denominator. Nineteen patients with WG died and 10 patients with MPA died during the study period, resulting in a point prevalence for survivors at 31 December 2003 of 112 cases/million (95% CI 82-142) and 37 cases/million (95% CI 20-55), respectively. Using unmodified ACR criteria the 5-yr period and point prevalence for WG were 131/million (95% CI 99-163) and 93.5/million (95% CI 66-121), respectively. Apart from respiratory tract involvement, which formed part of the case definition of WG, organ involvement was similar in both diseases. CONCLUSION: The prevalence of WG and MPA in Canterbury is the highest reported to date. Restricting the case definition of WG to the ACR classification criteria we found a prevalence equivalent to that described in northern Norway. The clinical severity and serological characteristics were similar to descriptions in other WG and MPA patient cohorts. Studies of disease prevalence in other Southern Hemisphere centres will determine if the observed north-south negative disease gradient in the Northern Hemisphere is reciprocated. PMID- 16368730 TI - The prevalence of headache in Behcet's syndrome. AB - OBJECTIVES: Behcet's syndrome is an uncommon systemic disorder that involves the nervous system in 5% of cases. Headache may arise in conjunction with such complications but also appears to occur independently. We sought to define the prevalence of headache in an unselected group of patients with Behcet's syndrome, to characterize the headache syndromes and to identify what treatments are being used. METHODS: A questionnaire was sent to an unselected group of patients through their support group newsletter. RESULTS: The results showed a prevalence of recurrent headache in 82.5% of responders; the majority exhibited symptoms that fulfilled the International Headache Society criteria for migraine, with a higher than normal prevalence of visual or sensory aura of 52%. Using the Migraine Disability Assessment (MIDAS) score for disability in migraine, 62% of responders showed moderate or severe disability. Headache treatment was poor, the majority of sufferers resorting to over-the-counter remedies; preventative treatments had rarely been prescribed. CONCLUSIONS: Recurrent headache is very common in Behcet's syndrome, is poorly treated and is associated with disablement. PMID- 16368731 TI - Acupuncture for osteoarthritic pain: an observational study in routine care. AB - OBJECTIVE: To investigate characteristics and outcomes of patients undergoing acupuncture treatment for osteoarthritic pain under conditions of routine care in the framework of statutory health insurance in Germany. METHODS: Patients with chronic pain due to osteoarthritis (ICD-10 diagnoses M15 to M19) treated with acupuncture as the leading form of therapy were included in an observational study. Detailed questionnaires including instruments to measure pain intensity (numerical rating scales from 0 to 10), disability (Pain Disability Index) and quality of life (SF-36) were filled in before treatment, after treatment and at 6 months. Patients suffering from osteoarthritis of the knee and hip also filled in the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index questionnaire. RESULTS: A total of 736 patients were included in the main analysis. Seventy (10%) patients and 278 (38%) patients, respectively, suffered exclusively from primary osteoarthritis of the hip or knee, 239 (33%) from another type of osteoarthritis and 149 (20%) had more than one affected joint. On average, patients received 8.7 +/- 3.1 acupuncture treatments. Statistically significant and clinically relevant improvements were seen in all subgroups both after treatment and at 6 months in all major outcome measures. In patients with osteoarthritis of the hip, the WOMAC sum score was 47.9 +/- 20.7 at baseline, 34.8 +/- 20.0 after treatment and 33.1 +/- 22.2 at 6 months. The respective values in patients with osteoarthritis of the knee were 51.7 +/- 20.9, 34.1 +/- 23.3 and 34.6 +/- 25.1. CONCLUSIONS: In this study, patients with chronic pain due to osteoarthritis reported clinically relevant improvements after acupuncture treatment. Due to the uncontrolled design and the high proportion of patients lost to follow-up, the study findings must be interpreted cautiously. PMID- 16368732 TI - Juvenile localized scleroderma: clinical and epidemiological features in 750 children. An international study. AB - OBJECTIVE: Juvenile localized scleroderma (JLS) includes a number of conditions often grouped together. With the long-term goal of developing uniform classification criteria, we studied the epidemiological, clinical and immunological features of children with JLS followed by paediatric rheumatology and dermatology centres. METHODS: A large, multicentre, multinational study was conducted by collecting information on the demographics, family history, triggering environmental factors, clinical and laboratory features, and treatment of patients with JLS. RESULTS: Seven hundred and fifty patients with JLS from 70 centres were enrolled into the study. The disease duration at diagnosis was 18 months. Linear scleroderma (LS) was the most frequent subtype (65%), followed by plaque morphea (PM) (26%), generalized morphea (GM) (7%) and deep morphea (DM) (2%). As many as 15% of patients had a mixed subtype. Ninety-one patients (12%) had a positive family history for rheumatic or autoimmune diseases; 100 (13.3%) reported environmental events as possible trigger. ANA was positive in 42.3% of the patients, with a higher prevalence in the LS-DM subtype than in the PM-GM subtype. Scl70 was detected in the sera of 3% of the patients, anticentromere antibody in 2%, anti-double-stranded DNA in 4%, anti-cardiolipin antibody in 13% and rheumatoid factor in 16%. Methotrexate was the drug most frequently used, especially during the last 5 yr. CONCLUSION: This study represents the largest collection of patients with JLS ever reported. The insidious onset of the disease, the delay in diagnosis, the recognition of mixed subtype and the better definition of the other subtypes should influence our efforts in educating trainees and practitioners and help in developing a comprehensive classification system for this syndrome. PMID- 16368733 TI - Preventing non-steroidal anti-inflammatory drug-induced gastrointestinal toxicity: are older strategies more cost-effective in the general population? AB - OBJECTIVES: To assess the relative cost-effectiveness of five gastroprotective strategies for patients in the general population not judged to be at high gastrointestinal (GI) risk requiring regular traditional (t) non-steroidal anti inflammatory drugs (NSAIDs) for over 3 weeks: tNSAID/H(2) receptor antagonists (H(2)RAs); tNSAID/proton pump inhibitors (PPIs); tNSAID/misoprostol; COX-2 preferential NSAIDs or COX-2-specific NSAIDs (COXIBs). METHODS: A systematic review of outcomes and UK cost data were combined in an incremental economic analysis. Incremental cost-effectiveness ratios were generated for quality adjusted life years (QALYs) gained. RESULTS: Cost-utility analysis showed a tNSAID with a H(2)RA is safer and less costly than tNSAIDs alone, and equally effective and less costly than COXIBs. tNSAID/misoprostol was also dominated by tNSAID/H(2)RA due to withdrawal caused by side-effects reducing overall health status. The incremental increase in QALYs gained by using COXIBs instead of tNSAID/H(2)RA would cost 670,000 pounds per QALY gained. The incremental increase in QALYs gained by using tNSAID/PPI instead of COXIBs would cost 26,000 pounds per QALY gained. If the decision-maker will pay up to 140,000 pounds per extra QALY, the optimal strategy is tNSAID/H(2)RA. If the decision-maker will pay over this the optimal strategy is tNSAID/PPI. CONCLUSION: The economic analysis suggests that there may be a case for prescribing H(2)RAs in all patients requiring NSAIDs. Our recommendations are tentative due to the quality of the data available and the assumptions we have had to make in our model, and it is possible that other strategies may be preferred in patients with higher baseline GI risk. PMID- 16368734 TI - Limited VH gene usage in B-cell clones established with nurse-like cells from patients with rheumatoid arthritis. AB - OBJECTIVES: Nurse-like stromal cells (NLC) in synovia and bone marrow of patients with rheumatoid arthritis (RA) can support pseudoemperipolesis, protect from apoptosis and enhance immunoglobulin production of peripheral blood B cells isolated from healthy individuals, suggesting the profound contribution of hyperactivation of B cells in RA. In the course of establishing RA-NLC from RA patients, we observed the growth of B cells in the presence of RA-NLC. METHODS: We cloned B cells from the synovium or bone marrow of RA patients using the limiting dilution technique. For established clones, nucleotide sequences of immunoglobulin and surface antigens were investigated. To investigate the dependence of these clones on NLC, differences in the proliferation and the amount of immunoglobulin produced in the presence or absence of NLC were compared. Immunocytochemical staining of various cells was performed using the antibody these clones produced. RESULTS: Nine B-cell clones established from RA patients showed RA-NLC-dependent growth. These B-cell clones expressed CD19, CD20, CD38, CD39 and CD40, suggesting that the cloned cells were mature and activated. All clones secreted immunoglobulins in culture media, which were specific for intracellular components of various cell lines, including RA-NLC. Interestingly, we found limited usage of immunoglobulin heavy-chain variable regions (VH) among B-cell clones from RA patients. These repertoires were reported to be detected preferentially in fetal livers. CONCLUSION: The present study provides a novel insight into the involvement of RA-NLC in the immunopathogenesis of RA via an autoreactive B cell development and/or activation mechanism. PMID- 16368735 TI - Diagnosing Kawasaki syndrome. PMID- 16368736 TI - Use of the 'STRATIFY' falls risk assessment in patients recovering from acute stroke. AB - OBJECTIVES: To investigate the predictive validity and reliability of the STRATIFY falls risk assessment tool as applied to patients recovering from acute stroke. DESIGN: Prospective cohort study. SETTING: Six stroke rehabilitation units in the North of England. SUBJECTS: All patients with a diagnosis of acute stroke admitted to the participating stroke units during a 6-month study period. ASSESSMENT: on admission, falls risk (STRATIFY), disability (Barthel index), mobility (Rivermead mobility index), cognitive impairment (abbreviated mental test score) and visual neglect (Albert's test) were assessed. Then, STRATIFY was completed weekly and within 48 h of anticipated discharge. Consenting patients were contacted at 3 months after discharge to determine falls. OUTCOME MEASURES: Occurrence of a fall within 28 days of the baseline STRATIFY (in-patient study), falls in the first 3 months after discharge (post-discharge study) and falls during stroke unit stay (reliability study). RESULTS: From 387 patients admitted to the participating units during the study period, 225 contributed to the 28 day in-patient study, and 234 were followed up at 3 months after discharge. STRATIFY performed poorly in predicting falls in the first 28 days (sensitivity 11.3% and specificity 89.5%) and after discharge (sensitivity 16.3% and specificity 86.4%). Agreement was 'fair' between baseline and discharge scores (kappa = 0.263) and 'good' between the pre-hospital discharge score and that obtained in the week preceding discharge (kappa = 0.639). CONCLUSION: STRATIFY performed poorly as a predictor of falls in a heterogeneous population of stroke patients. There is a need for a disease-specific rather than a generic falls risk assessment tool. PMID- 16368737 TI - Role of depressive and cognitive status in self-reported evaluation of quality of life in older people: comparing proxy and physician perspectives. AB - OBJECTIVE: To assess the quality of life (QOL) of older adults aged over 65 years, who were healthy or suffering from depressive syndrome (DS) and/or Alzheimer's disease (AD); to analyse agreement between participants' and proxies' QOL ratings; to evaluate the association between participants' depressive and cognitive symptoms and QOL rating; to correlate participants' health ratings and the severity of physician assessment. METHODS: 138 non-institutionalised older people of both genders and their respective caregiver and treating doctor were consecutively recruited (response rate 74.6%). Forty suffered from AD, 36 from DS, 35 from both conditions and 27 had neither. All participants were evaluated by Mini Mental State Examination, Geriatric Depression Scale and World Health Organization Quality of Life (WHOQOL) questionnaire. The caregiver filled out QOL Proxy and the physician filled out the 'Health and Severity of Illness' form. RESULTS: The four groups scored significantly differently in all areas of WHOQOL 100 (WHOQOL questionnaire with 100 items). Participants with DS perceived their QOL as poorer than did healthy and AD subjects. Participants with AD and DS obtained intermediate scores. Severity of depression correlated with worsening QOL. Subjects with DS--but not those with AD, AD and DS, and, in some areas, healthy participants--had similar perception of their QOL to their proxies. Poor physical health ratings by the physician corresponded to poorly perceived QOL by the patient. CONCLUSION: Older people with AD perceive their own QOL similarly to and, in some areas, even better than healthy people of the same age. The opposite was observed among the depressed. Informants do not always evaluate QOL in the same way as healthy elders and those with AD, while there is more agreement with depressed patients. Informant evaluation may be helpful but is not necessarily reliable. PMID- 16368738 TI - Functional influence of N-glycosylation in OCT2-mediated tetraethylammonium transport. AB - OCT2, an organic cation transporter critical for removal of many drugs and toxins from the body, contains consensus sites for N-glycosylation at amino acid position 71, 96, and 112. However, the extent to which these sites are glycosylated by the cell, and the influence glycosylation has on OCT2 function, remains unknown. To address these issues, the acquisition of N-glycosylation was disrupted by mutating the amino acid asparagine (N) to glutamine (Q) at these sites in the rabbit ortholog of OCT2, which was expressed in Chinese hamster ovary cells. Disruption of N-glycosylation followed by Western blotting indicated that each site is indeed glycosylated and that OCT2 contains no other sites of N glycosylation. Plasma membrane expression (determined by surface biotinylation) of the N112Q mutant, but not N71Q or N96Q mutants, was fourfold lower than that of wild-type OCT2, and unglycosylated OCT2 (N71Q/N96Q/N112Q) was sequestered in an unidentified intracellular compartment. The N71Q, N96Q, and N112Q mutants had a higher affinity ( approximately 2-fold) for tetraethylammonium (TEA). Maximum transport rate was reduced in the N96Q (3-fold) and N112Q (5-fold) mutants, but not the N71Q mutant, and unglycosylated OCT2 failed to transport TEA (associated with its absence in the plasma membrane). Whereas the reduction in maximum transport rate of the N112Q mutant is consistent with its reduced plasma membrane expression, the lower rate of the N96Q mutant, which appeared to traffic properly, suggests that glycosylation at N96 increases the transporter turnover number. PMID- 16368739 TI - TGF-beta1-induced EMT can occur independently of its proapoptotic effects and is aided by EGF receptor activation. AB - Apoptosis and epithelial-mesenchymal transdifferentiation (EMT) occur in stressed tubular epithelial cells and contribute to renal fibrosis. Transforming growth factor (TGF)-beta(1) promotes these responses and we examined whether the processes were interdependent in vitro. Direct (caspase inhibition) and indirect [epidermal growth factor (EGF) receptor stimulation] strategies were used to block apoptosis during TGF-beta(1) stimulation, and the subsequent effect on EMT was assessed. HK-2 cells were exposed to TGF-beta(1) with or without preincubation with ZVAD-FMK (pan-caspase inhibitor) or concomitant treatment with EGF plus or minus preincubation with LY-294002 (PI3-kinase inhibitor). Cells were then assessed for apoptosis and proliferation by flow cytometry, crystal violet assay, and Western blotting. Markers of EMT were assessed by microscopy, immunofluorescence, real-time RT-PCR, Western blotting, PAI-1 reporter assay, and collagen gel contraction assay. TGF-beta(1) caused apoptosis and priming for staurosporine-induced apoptosis. This was blocked by ZVAD-FMK. However, ZVAD-FMK did not prevent EMT following TGF-beta(1) treatment. EGF inhibited apoptosis and facilitated TGF-beta(1) induction of EMT by increasing proliferation and accentuating E-cadherin loss. Additionally, EGF significantly enhanced TGF beta(1)-induced collagen I gel contraction. EGF increased Akt phosphorylation during EMT, and the prosurvival effect of this was confirmed using LY-294002, which reduced EGF-induced Akt phosphorylation and reversed its antiapoptotic and proproliferatory effects. TGF-beta(1) induces EMT independently of its proapoptotic effects. TGF-beta(1) and EGF together lead to EMT. EGF increases proliferation and resistance to apoptosis during EMT in a PI3-K Akt-dependent manner. In vivo, EGF receptor activation may assist in the selective survival of a transdifferentiated, profibrotic cell type. PMID- 16368740 TI - Keratinocyte-derived chemokine is an early biomarker of ischemic acute kidney injury. AB - Renal ischemia-reperfusion injury (IRI) is the leading cause of acute kidney injury [AKI; acute renal failure (ARF)] in native kidneys and delayed graft function in deceased donor kidney transplants. Serum creatinine rises late after renal IRI, which results in delayed diagnosis. There is an important need to identify novel biomarkers for early diagnosis and prognosis in renal IRI. Given the inflammatory pathophysiology of renal IRI, we used a protein array to measure 18 cytokines and chemokines in a mouse model of renal IRI at 3, 24, and 72 h postischemia. A rise in renal keratinocyte-derived chemokine (KC) was the earliest and most consistent compared with other molecules, with 3-h postischemia values being 9- and 13-fold greater than sham and normal animals, respectively. Histological changes were evident within 1 h of IRI but serum creatinine only increased 24 h after IRI. With the use of an ELISA, KC levels in serum and urine were highest 3 h postischemia, well before a significant rise in serum creatinine. The human analog of KC, Gro-alpha, was markedly elevated in urine from humans who received deceased donor kidney transplants that required dialysis, compared with deceased donor kidney recipients with good graft function and live donor recipients with minimal ischemia. Measurement of KC and its human analog, Gro-alpha, could serve as a useful new biomarker for ischemic ARF. PMID- 16368741 TI - Biomechanical characterization of the urethral musculature. AB - Rigorous study of the associations between urethral structural anatomy and biomechanical function is necessary to advance the understanding of the development, progression, and treatment of urethral pathologies. An ex vivo model was utilized to define the relative biomechanical contributions of the active (muscle) elements of the female urethra relative to its passive (noncontractile) elements. Whole urethras from female, adult rats were tested under a range of applied intraluminal pressures (0 to 20 mmHg) as a laser micrometer simultaneously measured midurethral outer diameter. Active tissue characterization was performed during induced contraction of either smooth muscle alone (N(omega)-nitro-l-arginine, phenylephrine), striated muscle alone (sodium nitroprusside, atropine, hexamethonium, acetylcholine), or during collective activation of both muscles (N(omega)-nitro-l-arginine, phenylephrine, acetylcholine). The subsequent collection of paired passive biomechanical responses permitted the determination of parameters related to intrinsic muscle contractile function. Activation of each muscle layer significantly influenced the biomechanical responses of the tissue. Measures of muscle responsiveness over a wide range of sustained opposing pressures indicated that an activated striated muscle component was approximately one-third as effective as activated smooth muscle in resisting tissue deformation. The maximum circumferential stress generated by the striated muscle component under these conditions was also determined to be approximately one-third of that generated by the smooth muscle (748 +/- 379 vs. 2,229 +/- 409 N/m(2)). The experiments quantitatively reveal the relative influence of the intrinsic urethral smooth and striated muscle layers with regard to their effect on the mechanical properties and maximum functional responses of the urethra to applied intralumenal stresses in the complete absence of extrinsic influences. PMID- 16368742 TI - Glycosylation of the osmoresponsive transient receptor potential channel TRPV4 on Asn-651 influences membrane trafficking. AB - We identified a consensus N-linked glycosylation motif within the pore-forming loop between the fifth and sixth transmembrane segments of the osmoresponsive transient receptor potential (TRP) channel TRPV4. Mutation of this residue from Asn to Gln (i.e., TRPV4(N651Q)) resulted in loss of a slower migrating band on anti-TRPV4 immunoblots and a marked reduction in lectin-precipitable TRPV4 immunoreactivity. HEK293 cells transiently transfected with the mutant TRPV4(N651Q) exhibited increased calcium entry in response to hypotonic stress relative to wild-type TRPV4 transfectants. This increase in hypotonicity responsiveness was associated with an increase in plasma membrane targeting of TRPV4(N651Q) relative to wild-type TRPV4 in both HEK293 and COS-7 cells but had no effect on overall channel abundance in whole cell lysates. Residue N651 of TRPV4 is immediately adjacent to the pore-forming loop. Although glycosylation in this vicinity has not been reported for a TRP channel, the structurally related hexahelical hyperpolarization-activated cyclic nucleotide-gated channel, HCN2, and the voltage-gated potassium channel, human ether-a-go-go-related (HERG), share a nearly identically situated and experimentally confirmed N-linked glycosylation site which promotes rather than limits channel insertion into the plasma membrane. These data point to a potentially conserved structural and functional feature influencing membrane trafficking across diverse members of the voltage-gated-like ion channel superfamily. PMID- 16368743 TI - Liver X receptor agonist TO-901317 upregulates SCD1 expression in renal proximal straight tubule. AB - Liver X receptors (LXRs), including LXRalpha and LXRbeta, are intracellular sterol sensors that regulate expression of genes controlling fatty acid and cholesterol absorption, excretion, catabolism, and cellular efflux. Because the kidney plays an important role in lipid metabolism and dyslipidemia accelerates renal damage, we investigated the effect of TO-901317, an LXR agonist, on the gene expression profile in mouse kidney. Treatment of C57 Bl/6 mice with TO 901317 (3 mg.kg(-1).day(-1)) for 3 days resulted in 51 transcripts that were significantly regulated in the kidney. Among them, the stearoyl-CoA desaturase-1 (SCD1) was upregulated most dramatically. Northern blot analysis revealed that SCD1 mRNA levels were markedly higher than that in control kidneys. Enhanced SCD1 expression by TO-901317 also resulted in increased fatty acid desaturation in the kidney. In control mice, constitutive renal SCD1 expression was low; however, TO 901317 treatment markedly increased SCD1 expression in the outer stripe of the outer medulla as assessed by both in situ hybridization and immunostain. Double labeling studies further indicated that SCD1 mRNA was selectively expressed in proximal straight tubules negative for aquaporin-2 and Tamm-Horsfall protein. In vitro studies in cultured murine proximal tubule cells further demonstrated that LXR activation enhanced SCD1 transcription via increased sterol regulatory element binding protein-1. Taken together, these data suggest LXR activation of SCD1 expression may play an important role in regulating lipid metabolism and cell function in renal proximal straight tubules. PMID- 16368744 TI - Effects of a single administration of acarbose on postprandial glucose excursion and endothelial dysfunction in type 2 diabetic patients: a randomized crossover study. AB - CONTEXT: Postprandial hyperglycemia has been reported to elicit endothelial dysfunction and provoke future cardiovascular complications. A reduction of postprandial blood glucose levels by the alpha-glucosidase inhibitor acarbose was associated with a risk reduction of cardiovascular complications, but effects of acarbose on endothelial function have never been elucidated. DESIGN: This study was aimed to assess the efficacy of acarbose on postprandial metabolic parameters and endothelial function in type 2 diabetic patients. Postprandial peakglucose (14.47 +/- 1.27 vs. 8.50 +/- 0.53 mmol/liter), plasma glucose excursion (PPGE), and change in the area under the curve (DeltaAUC)glucose after a single loading of test meal (total 450 kcal; protein 15.3%; fat 33.3%; carbohydrate 51.4%) were significantly higher in diet-treated type 2 diabetic patients (n = 14) than age- and sex-matched controls (n = 12). RESULTS: The peak forearm blood flow response and total reactive hyperemic flow (flow debt repayment) during reactive hyperemia, indices of resistance artery endothelial function on strain-gauge plethysmography, were unchanged before and after meal loading in controls. But those of diabetics were significantly decreased 120 and 240 min after the test meal. A prior administration of acarbose decreased postprandial peakglucose, PPGE, and DeltaAUCglucose. The peak forearm blood flow and flow debt repayment were inversely well correlated with peakglucose, PPGE, and DeltaAUCglucose but not with DeltaAUCinsulin or the other lipid parameters. CONCLUSIONS: Even a single loading of test meal was shown to impair endothelial function in type 2 diabetic patients, and the postprandial endothelial dysfunction was improved by a prior use of acarbose. Acarbose might reduce macrovascular complication by avoiding endothelial injury in postprandial hyperglycemic status. PMID- 16368745 TI - Serum anti-Mullerian hormone as a surrogate for antral follicle count for definition of the polycystic ovary syndrome. AB - CONTEXT: Despite its frequency, the polycystic ovary syndrome (PCOS) is still a difficult diagnosis in endocrinology, gynecology, and reproductive medicine. To help solve this issue, the Rotterdam consensus conference proposed to include the ultrasonographic follicle count as a new diagnostic criterion, in addition to hyperandrogenism and oligo-anovulation. Unfortunately, its assessment does not offer sufficient reliability worldwide. OBJECTIVE: The aim of our study was to check whether anti-Mullerian hormone (AMH) measurement in the serum could be a surrogate for antral follicle count in the diagnostic criteria of PCOS. DESIGN, SETTING, AND PATIENTS: Serum AMH was measured with a second-generation immunoassay in a cohort of 73 PCOS patients and 96 controls, and its diagnostic power was evaluated by receiver operating characteristic curves. PCOS was diagnosed according to the Rotterdam definition. RESULTS: Serum AMH levels were 3 fold higher in PCOS patients than in controls (81.6 vs. 33.5 pmol/liter; P < 0.001) and were significantly related to the follicle number in the two groups. The area under the receiver operating characteristic curve for the AMH assay was 0.851, indicating a good diagnostic potency. Setting the threshold at 60 pmol/liter offered the best compromise between specificity (92%) and sensitivity (67%). CONCLUSIONS: The serum AMH level is an accurate marker of the ovarian early antral follicle number and offers a good diagnostic potency. In situations where accurate ultrasonographic data are not available, AMH could thus be used instead of the follicle count as a diagnostic criterion and incorporated as such in the Rotterdam definition of PCOS. PMID- 16368746 TI - Follicle-stimulating hormone-independent functions of primate Sertoli cells: potential implications in the diagnosis and management of male infertility. AB - CONTEXT: FSH is known to augment the production of essential germ cell (Gc) survival factors, lactate and estradiol, by Sertoli cells (Sc) of 18-d-old pubertal rats. However, the failure of gonadotropin and androgen treatment to initiate spermatogenesis in testis of some infertile men bearing Sc and Gc is intriguing. The role of FSH in regulation of lactate and estradiol production by primate Sc is currently unknown. OBJECTIVE: The objective of the study was to determine the role of FSH in regulating lactate and estradiol production by primate Sc. METHODS: Gc differentiation was initiated in male juvenile rhesus monkeys by pulsatile administration of GnRH for 4-5 wk. Sc from these pseudopubertal monkeys and pubertal rats were cultured. Production of lactate and estradiol in response to FSH and 8-bromoadenosine-cAMP was evaluated. Inhibin betaB expression, cAMP production, and cell proliferation were also assayed. RESULTS: Unlike Sc from pubertal rats, Sc from pseudopubertal monkeys constitutively aromatized testosterone to estradiol and produced large amounts of lactate without FSH stimulation. Increasing doses of recombinant monkey FSH or 8 bromoadenosine-cAMP failed to augment lactate production, although they significantly augmented proliferation of Sc. Production of cAMP and expression of inhibin-betaB mRNA were also remarkably augmented by recombinant monkey FSH. CONCLUSIONS: These results suggest that lactate and estradiol production by monkey Sc is not governed by FSH, as previously thought based on studies of rat Sc. Thus, in a clinical situation, assessment of such gonadotropin-independent functions of Sc may be obligatory for the diagnosis and management of certain forms of idiopathic male infertility. PMID- 16368747 TI - Hormonal and biochemical determinants of trabecular microstructure at the ultradistal radius in women and men. AB - CONTEXT: Using high-resolution three-dimensional peripheral quantitative computed tomography (3D-pQCT) imaging, we recently described sex and age effects on bone microstructure at the ultradistal radius in men and women. Although bone volume/tissue volume decreased with age in both sexes, changes in trabecular number (TbN) and thickness (TbTh) in men were complex, with evidence for conversion of thick trabeculae into more numerous, thinner trabeculae in young men. OBJECTIVE: Our objective was to define the relationship between hormonal and bone turnover variables and trabecular microstructure at the ultradistal radius. DESIGN, SETTING, AND PARTICIPANTS: We conducted a population-based, cross sectional study in the general community that included 205 women and 269 men, aged 21-97 yr. MAIN OUTCOME MEASURES: We measured correlation of bone volume/tissue volume, TbN, TbTh, and trabecular separation with hormonal and bone turnover variables. RESULTS: In young men (20-39 yr), TbTh and TbN were associated with serum IGF-I levels (r = 0.31, P < 0.05 and r = -0.35, P < 0.01, respectively). No associations were found between sex steroid levels (bioavailable estradiol or testosterone) or biochemical markers of bone turnover and trabecular parameters in young men or women. By contrast, in elderly men and women (>60 yr), sex steroids were the most consistently associated with trabecular microstructure, and bone turnover markers were variably associated with these parameters. CONCLUSIONS: In young men, the apparent conversion of thick trabeculae into more numerous, thinner trabeculae is most closely associated with declining IGF-I levels. By contrast, sex steroids are the major hormonal determinants of trabecular microstructure in elderly men and women. PMID- 16368748 TI - Polymorphisms in the CLCN7 gene modulate bone density in postmenopausal women and in patients with autosomal dominant osteopetrosis type II. AB - CONTEXT: Genetic factors are important determinants of bone mineral density (BMD). The fact that mutations in the ClC-7 chloride channel cause autosomal dominant osteopetrosis (ADOII) make the CLCN7 gene an attractive candidate for the regulation of bone density. OBJECTIVE: The objective of the study was to investigate the association between polymorphisms in the CLCN7 gene and BMD in postmenopausal women and with clinical variability in ADOII. DESIGN: This was a genetic association study using five single-nucleotide polymorphisms and a variable number tandem repeat (VNTR) polymorphism in the CLCN7 gene. PARTICIPANTS: A total of 425 postmenopausal women aged 64 +/- 7 yr participated in the study. We also investigated an ADOII family with low penetrance comprising 18 mutation carriers. MAIN OUTCOME MEASURE(S): In our postmenopausal cohort, individual single-nucleotide polymorphism genotypes and haplotypes were analyzed for association with BMD at the lumbar spine and the femoral neck and with the bone resorption marker deoxypyridinoline (D-Pyr/Crea). The same polymorphisms on the nonmutated CLCN7 allele were investigated for association with the variability of the ADOII phenotype. RESULTS: Analysis by multiple linear regression revealed a significant association between the ss genotype of the VNTR and higher Z-score values (P = 0.029). The haplotype 4, which comprises the long allele of the VNTR, was found to be significantly associated with lower femoral neck Z-score values (P = 0.011). Furthermore, we found an association of the ss genotype of the VNTR with lower levels of the bone resorption marker D-Pyr/Crea (P = 0.015), whereas haplotype 4 was associated with higher D-Pyr/Crea levels (P = 0.039). In the ADOII family, we could demonstrate that haplotype 3, which contains the s-allele of the VNTR, is associated with a slightly higher probability that mutation carriers develop osteopetrosis (P = 0.029). In both cases the association seems largely to be driven by the VNTR genotype but is further strengthened if surrounding polymorphisms are added to the analysis. CONCLUSION: We observed a significant association of CLCN7 polymorphisms with the variance of BMD and bone resorption marker levels in postmenopausal women and with the variability of the ADOII phenotype. PMID- 16368749 TI - High-density lipoprotein (HDL) transport in the metabolic syndrome: application of a new model for HDL particle kinetics. AB - CONTEXT: Reduced high density lipoprotein (HDL) concentration in the metabolic syndrome (MetS) is associated with increased risk of diabetes and cardiovascular disease and is related to defects in the kinetics of HDL apolipoprotein (apo) A-I and A-II. OBJECTIVE: The objective of the study was to investigate HDL apoA-I and apoA-II kinetics in nondiabetic men with MetS and lean controls by developing a model that describes the kinetics of lipoprotein (Lp)A-I and LpA-I:A-II particles. DESIGN: Twenty-three MetS men and 10 age-matched lean controls were investigated. ApoA-I and apoA-II tracer/tracee ratios were studied after iv d3 leucine administration using gas chromatography mass spectrometry. RESULTS: Compared with lean subjects, MetS subjects had accelerated catabolism of LpA-I (P < 0.001), LpA-I:A-II (P = 0.005), and apoA-II (P = 0.005); the production rate of LpA-I was also significantly elevated in MetS, so that the dominant changes in plasma concentrations were reduction in LpA-I:A-II (P < 0.001) and apoA-II (P < 0.05). Increased catabolism of LpA-I and LpA-I:A-II was directly related to increased waist circumference, hypertriglyceridemia, low HDL-cholesterol, small HDL particle size, hyperinsulinemia, and low phospholipid transfer protein (PLTP) activity; overproduction of LpA-I was significantly associated with increased waist circumference, insulin resistance, and low PLTP activity. CONCLUSIONS: MetS men exhibit hypercatabolism of the two major HDL lipoprotein particles, LpA-I and LpA-I:A-II, but selective overproduction of LpA-I maintains a normal plasma concentration of LpA-I. These kinetic perturbations are probably related to central obesity, insulin resistance, hypertriglyceridemia, and low plasma PLTP activity. PMID- 16368750 TI - Testosterone and estradiol among older men. AB - CONTEXT: Testosterone and estradiol levels decline with age in men. This change may affect multiple clinical outcomes, but there have been few reports of the distribution and correlates of testosterone and estradiol concentrations in elderly men. OBJECTIVE: The purpose of these studies was to assess sex steroid levels in a large cohort of older men. DESIGN: We conducted a cross-sectional cohort evaluation. SETTING: Community-dwelling men were studied at six academic medical centers in the United States. PARTICIPANTS: The Osteoporotic Fractures in Men Study is a prospective cohort of men aged at least 65 yr. In these studies, a randomly selected stratified subsample of 2623 participants was analyzed. MAIN OUTCOME MEASURES: We assessed levels of total and free testosterone and estradiol and SHBG. RESULTS: Age was inversely associated with free testosterone and free estradiol levels (P for trend = 0.001 for both). Notably, at any age, there was substantial variation in levels of each hormone. Free testosterone levels were lower in men with greater body mass index, lower SHBG, and poorer self-reported health status and in those of Asian race. Free estradiol concentrations were lower in men with lower body mass index and higher SHBG levels. Free estradiol and free testosterone were modestly correlated (r = 0.20; P < 0.001), but at any level of free testosterone, there was considerable variation in free estradiol levels. CONCLUSIONS: This is the largest cohort of older men in which sex steroid levels are available, and it demonstrates that testosterone and estradiol, and their free fractions, tend to decline with age even among older men. However, substantial variation is also present. The relationships between sex steroid levels and their consequences in aging are likely to be complex. PMID- 16368751 TI - A novel deletion in the GH1 gene including the IVS3 branch site responsible for autosomal dominant isolated growth hormone deficiency. AB - CONTEXT: The majority of mutations responsible for isolated GH type II deficiency (IGHD II) lead to dominant negative deleteriously increased levels of the GH1 exon 3 skipped transcripts. OBJECTIVE: The aim of this study was the characterization of the molecular defect causing a familial case of IGHD II. PATIENTS: A 2-yr-old child and her mother with severe growth failure at diagnosis (-5.8 and -6.9 sd score, respectively) and IGHD were investigated for the presence of GH1 mutations. RESULTS: We identified a novel 22-bp deletion in IVS3 (IVS3 del+56-77) removing the putative branch point sequence (BPS). Analysis of patients' lymphocyte mRNA showed an excess exon 3 skipping. The mutated allele transfected into rat pituitary cells produced four differently spliced products: the exon 3 skipped mRNA as the main product and lower amounts of the full-length cDNA and of two novel mRNA aberrant isoforms, one with the first 86 bases of exon 4 deleted and the other lacking the entire exon 4. A mutagenized construct lacking exclusively the 7 bp of the BPS only generated the exon 4 skipped and the full-length isoforms. The presence of the full-length transcript in the absence of the canonical BPS points to an alternative BPS in IVS3. CONCLUSION: The IVS3 del+56-77 mutation, causing IGHD II in this family, has two separate effects on mRNA processing: 1) exon 3 skipping, analogous to most described cases of IGHD II, an effect likely caused by the reduction in size of the IVS3, and 2) partial or total exon 4 skipping, as a result of the removal of the BPS. PMID- 16368752 TI - Growth hormone (GH) substitution in GH-deficient patients inhibits 11beta hydroxysteroid dehydrogenase type 1 messenger ribonucleic acid expression in adipose tissue. AB - CONTEXT: Local tissue activity of glucocorticoids is in part determined by the isoenzymes 11beta-hydroxysteroid dehydrogenase 1 (11beta-HSD1) and 11beta-HSD2, interconverting inert cortisone and active cortisol. Increased tissue activity of cortisol may play a central role in the features of GH deficiency and the metabolic syndrome. OBJECTIVE: We investigated the effects of GH treatment on adipose tissue 11beta-HSD mRNA. SUBJECTS AND METHODS: A randomized placebo controlled double-blind study design was used. Twenty-three GH-deficient patients (16 males and seven females) were randomized to 4 months of GH treatment (2 IU/m2) (n = 11) or placebo treatment (n = 12). Adipose tissue biopsies and blood samples were obtained before and after treatment. Biopsies were obtained from the abdominal sc depot at the level of the umbilicus and do not necessarily reflect the metabolically more important visceral adipose tissue. Gene expressions were determined by real-time RT-PCR. RESULTS: GH treatment decreased 11beta-HSD1 mRNA 66% [95% confidence interval (CI), 23-107%; P < 0.01] and increased 11beta-HSD2 mRNA 167% (95% CI, 33-297%; P < 0.05) in adipose tissue. Serum IGF-I and IGF-I mRNA increased in the GH-treated group by 187% (95% CI, 122-250%; P < 0.001) and 470% (95% CI, 88-846%; P < 0.01). The change in 11beta-HSD1 mRNA expression was negatively correlated with the change in serum IGF-I (R = -0.434; P < 0.05). In contrast, the change in 11beta-HSD2 mRNA expression was positively correlated with the change in serum IGF-I (R = 0.487; P < 0.05), and even stronger with the change in IGF-I mRNA expression (R = 0.798; P < 0.0001). CONCLUSION: GH treatment is able to decrease 11beta-HSD1 mRNA and increase 11beta-HSD2 and accordingly may be able to reduce the amount of locally produced cortisol in adipose tissue. PMID- 16368753 TI - Diagnostic and prognostic value of 18-fluorodeoxyglucose positron emission tomography in adrenocortical carcinoma: a prospective comparison with computed tomography. AB - OBJECTIVE: Patients with adrenocortical cancer are submitted to multiple imaging procedures for diagnosis of recurrence and staging. The aim of this prospective study was to evaluate the diagnostic and prognostic values of fluorodeoxyglucose (FDG) using a combined positron emission tomography and computed tomography (PET/CT) modality, compared with thoracoabdominopelvic computed tomography (TAP CT). METHODS: Twenty-eight consecutive patients with adrenocortical cancer referred from November 2003 to December 2004 to the Institut Gustave Roussy were included. Mean time between PET/CT and TAP-CT was 16 d. Independent readers analyzed images of each modality. The gold standard was progression on follow-up TAP-CT or pathology. RESULTS: A total of 269 lesions in 57 organs were depicted in 22 patients. The sensitivities for the detection of distinct lesions and the diagnosis of metastatic organs were 90 and 93% for PET/CT and 88 and 82% for TAP CT, respectively. Twelve percent of the lesions were seen on PET/CT only and 10% on TAP-CT only. Eighteen percent of the metastatic organs were diagnosed with PET/CT only and 7% with TAP-CT only. Thirty-eight percent of the local relapses were seen only with PET/CT. PET/CT depicted three false-positive lesions. Treatment modalities were modified by PET/CT findings in five cases among which one was falsely positive. Tumor size and mitotic rate were significantly associated with FDG uptake. The intensity of FDG uptake (maximum standardized uptake value > 10) and the volume of FDG uptake (>150 ml) were significant prognostic factors for survival. CONCLUSIONS: We show that FDG-PET/CT is complementary to TAP-CT and of special interest in the diagnosis of local relapses. PMID- 16368754 TI - Interaction between testosterone and apolipoprotein E epsilon4 status on cognition in healthy older men. AB - CONTEXT: Reduced testosterone levels have been implicated as a potential causative factor in cognitive decline with older age. Men who possess the apolipoprotein E (APOE) epsilon4 allele have an increased risk of developing Alzheimer's disease; however, no studies have examined whether the influence of testosterone on cognition in healthy older men may be modulated by this genetic predisposition. OBJECTIVE: The objective of the study was to investigate the association between serum testosterone concentrations and cognitive performance in healthy older men, taking into account APOE epsilon4 status. DESIGN: This was a cross-sectional study conducted from 2003 to 2004. SETTING: The study population consisted of community-dwelling males residing in Perth, Western Australia. PARTICIPANTS: Healthy men over 55 yr, free of cognitive impairment and dementia (n = 45), were included in the study. MAIN OUTCOME MEASURES: Participants had fasting early morning blood samples for testosterone and SHBG and were assessed for mood as well as indices of general cognition, verbal and visual memory, executive functioning, working memory, and attention. RESULTS: There was a significant interaction between calculated free testosterone (FT) and APOE epsilon4 on general cognition (P = 0.01) and executive functioning, working memory, and attention (P < 0.01). Higher levels of FT were associated with better general cognition in non-epsilon4 carriers (P = 0.01). By contrast, in epsilon4 carriers higher FT levels were associated with lower scores on tests of executive functioning, working memory, and attention (P = 0.02). In men at increased risk for Alzheimer's disease, higher testosterone levels were not associated with better cognitive function. CONCLUSIONS: Cross-sectional and prospective studies of testosterone and cognition in older men should take into account APOE epsilon4 status. PMID- 16368755 TI - Role of CD95-mediated adipocyte loss in autoimmune lipodystrophy. AB - CONTEXT: Lipodystrophies are rare disorders characterized by the selective loss of adipose tissue. Metabolic complications increase in severity with the extent of fat loss. In some forms of acquired lipodystrophy, the loss of fat is suggested to be a result of autoimmune destruction of adipocytes. Here, the pathogenic mechanism is still poorly understood. OBJECTIVE: We have analyzed sc adipose tissue from a 5-yr-old girl with ongoing fat loss by immunohistochemistry. Using cultured human preadipocytes and adipocytes, we elucidated a possible mechanism leading to adipocyte loss in this patient. RESULTS: Analysis of adipose tissue samples of the patient with acquired lipodystrophy obtained from skin areas affected by panniculitis suggested that loss of adipocytes was mediated by CD95-induced apoptosis. Regression of adipose tissue was accompanied by lymphohistiocytic infiltration/inflammation and increased serum levels of inflammatory cytokines interferon-gamma and TNF-alpha. In vitro studies with human adipocytes demonstrated that interferon-gamma and TNF alpha are able to up-regulate CD95 expression and enhance CD95-death-inducing signaling complex formation resulting in a robust sensitization for CD95-mediated apoptosis. CONCLUSION: We have identified here a possible mechanism responsible for the loss of adipocytes by apoptosis in autoimmune lipodystrophy. PMID- 16368756 TI - A general binding mechanism for all human sulfatases by the formylglycine generating enzyme. AB - The formylglycine (FGly)-generating enzyme (FGE) uses molecular oxygen to oxidize a conserved cysteine residue in all eukaryotic sulfatases to the catalytically active FGly. Sulfatases degrade and remodel sulfate esters, and inactivity of FGE results in multiple sulfatase deficiency, a fatal disease. The previously determined FGE crystal structure revealed two crucial cysteine residues in the active site, one of which was thought to be implicated in substrate binding. The other cysteine residue partakes in a novel oxygenase mechanism that does not rely on any cofactors. Here, we present crystal structures of the individual FGE cysteine mutants and employ chemical probing of wild-type FGE, which defined the cysteines to differ strongly in their reactivity. This striking difference in reactivity is explained by the distinct roles of these cysteine residues in the catalytic mechanism. Hitherto, an enzyme-substrate complex as an essential cornerstone for the structural evaluation of the FGly formation mechanism has remained elusive. We also present two FGE-substrate complexes with pentamer and heptamer peptides that mimic sulfatases. The peptides isolate a small cavity that is a likely binding site for molecular oxygen and could host reactive oxygen intermediates during cysteine oxidation. Importantly, these FGE-peptide complexes directly unveil the molecular bases of FGE substrate binding and specificity. Because of the conserved nature of FGE sequences in other organisms, this binding mechanism is of general validity. Furthermore, several disease-causing mutations in both FGE and sulfatases are explained by this binding mechanism. PMID- 16368757 TI - Predicting microbial species richness. AB - Microorganisms are spectacularly diverse phylogenetically, but available estimates of their species richness are vague and problematic. For example, for comparable environments, the estimated numbers of species range from a few dozen or hundreds to tens of thousands and even half a million. Such estimates provide no baseline information on either local or global microbial species richness. We argue that this uncertainty is due in large part to the way statistical tools are used, if not indeed misused, in biodiversity research. Here we develop a powerful synthetic statistical approach to quantify biodiversity. It provides statistically sound estimates of microbial richness at any level of taxonomic hierarchy. We apply this approach to a large original 16S rRNA dataset on marine bacterial diversity and show that the number of bacterial species in a sample from marine sediments is (2.4 +/- 0.5 SE) x 10(3). We argue that our methodology provides estimates of microbial richness that are reliable and general, have biologically meaningful SEs, and meet other fundamental statistical standards. This approach can be an essential tool in biodiversity research, and the estimates of microbial richness presented here can serve as a baseline in microbial diversity studies. PMID- 16368758 TI - Loss of the limbic mineralocorticoid receptor impairs behavioral plasticity. AB - Corticosteroid action in the brain is mediated by the mineralocorticoid (MR) and the glucocorticoid (GR) receptor. Disturbances in MR- and GR-mediated effects are thought to impair cognition, behavior, and endocrine control. To assess the function of the limbic MR in these processes, we inactivated the MR gene in the forebrain of the mouse using the Cre/loxP-recombination system. We screened the mice with a limbic MR deficiency in various learning and exploration tests. The mutant mice show impaired learning of the water-maze task and deficits in measures of working memory on the radial maze due to behavioral perseverance and stereotypy. They exhibit a hyperreactivity toward a novel object but normal anxiety-like behavior. The behavioral changes are associated with abnormalities of the mossy fiber projection and an up-regulation of GR expression in the hippocampus. Adult mutant mice show normal corticosterone levels at circadian trough and peak. This genetic model provides important information about the consequences of a permanently altered balance between limbic MR and GR, with implications for stress-related neuroendocrine and neuropsychiatric diseases. PMID- 16368759 TI - The roles of victim and perpetrator alcohol use in intimate partner violence outcomes. AB - Alcohol use increases the risk of intimate partner violence (IPV), yet little research has examined its role in victimization outcomes (e.g., physical injury, police reporting). This study examined the roles of perpetrator and victim incident-specific alcohol use in IPV outcomes. The sample included 501 men and 1,756 women who had experienced an IPV physical assault. Logistic regression analyses showed that after controlling for relevant covariates, women whose partners had been drinking were significantly more likely to be injured than were women whose partners had not been drinking. A woman's own alcohol use was unrelated to victimization outcomes. Men were significantly more likely to report the incident if their partners had been drinking but were marginally less likely to report if they had been drinking. Findings suggest that perpetrators' incident specific alcohol use is important in understanding which victims are most likely to be injured and report the incident to the police. PMID- 16368760 TI - Impression management and self-report among violent offenders. AB - Offenders are assumed by many to employ socially desirable responding (SDR) response styles when completing self-report measures. Contrary to expectations, prior research has shown that accounting for SDR in self-report measures of antisocial constructs does not improve the relationship with outcome. Despite this, many self-report measures reliably predict future criminal outcome criteria. The present research examines the relationship of SDR (self-deception and impression management) with self-reported antisocial attitudes and the outcome of criminal recidivism in a sample of violent offenders. Offenders high on impression management reported lower antisocial attitudes. However, when impression management was statistically partialed from antisocial attitudes, the relationship with recidivism tended to diminish, though not to a statistically significant degree. This finding, though hypothesized based on previous empirical findings, is contrary to the theoretical assumption that controlling for SDR should improve the relationship of self-report with outcome. The discussion centers on the implications of routinely removing impression management from self report. PMID- 16368761 TI - Intimate partner violence and HIV/STD risk among lesbian, gay, bisexual, and transgender individuals. AB - To date, there has been little research examining HIV/STD risk among lesbian, gay, bisexual, and transgender (LGBT) individuals who are in abusive relationships. This article uses data collected from a community-based organization that provides counseling for LGBT victims of intimate partner violence (IPV). A total of 58 clients completed the survey, which inquired as to sexual violence and difficulties negotiating safer sex with their abusive partners. A large percentage of participants reported being forced by their partners to have sex (41%). Many stated that they felt unsafe to ask their abusive partners to use safer sex protection or that they feared their partners' response to safer sex (28%). In addition, many participants experienced sexual (19%), physical (21%), and/or verbal abuse (32%) as a direct consequence of asking their partner to use safer sex protection. Training counselors on issues of sexuality and safer sex will benefit victims of IPV. PMID- 16368762 TI - Changing coverage of domestic violence murders: a longitudinal experiment in participatory communication. AB - Stressing relation-building and participatory communication approaches, the Rhode Island Coalition against Domestic Violence worked with journalists to develop a best practices handbook on news coverage of domestic violence murders. This study compares print coverage of domestic violence murders prehandbook (1996-1999) and posthandbook (2000-2002). Significant changes include increased labeling of the murder of intimates as domestic violence and doubled usage of advocates as sources. As a result, domestic violence murders, previously framed as unpredictable private tragedies, are more commonly framed posthandbook as social problems warranting public intervention. The authors conclude that relation building approaches can affect news cultures and public discourse when conducted in conjunction with comprehensive participatory communications strategies. PMID- 16368763 TI - Feelings of revenge, retaliation motive, and posttraumatic stress reactions in crime victims. AB - Individuals with posttraumatic stress disorder (PTSD) are often said to experience strong feelings of revenge. However, there is a need for confirmatory empirical studies. Therefore, in a study of 174 victims of violent crimes, the relation between feelings of revenge and posttraumatic stress reactions was investigated. Feelings of revenge were correlated with intrusion and hyperarousal but not with avoidance. Feelings of revenge explained incremental variance of intrusion and hyperarousal when the variance explained by victimological variables was controlled. The retaliation motive implied in feelings of revenge did not account for the relation between feelings of revenge and posttraumatic stress reactions. However, the relation was moderated by the time since victimization. Therefore, feelings of revenge must presumably be regarded as a maladaptive coping reaction to experienced injustice, but not in the first period after victimization. PMID- 16368764 TI - Childhood physical punishment and problem solving in marriage. AB - Drawing from social learning theories and symbolic interactionist understandings of social life, the authors suggest that physical punishment teaches aggressive and controlling strategies for solving the problems of living together and hinders the development of important problem-solving skills, specifically the ability to role take with others. These strategies and skills become part of an individual's toolkit for problem resolution within his or her marriage. The analysis is based on 188 married couples in Washington State who participated in a longitudinal study of the first 2 years of marriage. The analysis reveals the following: Individuals who were physically punished during childhood are more likely to engage in physical and verbal aggression with their spouses, individuals who were physically punished during childhood are more controlling with their spouses, and individuals who were physically punished during childhood are less able to take their spouse's perspective. PMID- 16368765 TI - Intimate partner violence and long-term psychosocial functioning in a national sample of American women. AB - Using a nationally representative sample of American married or cohabiting women, this prospective study examined women who reported or denied intimate partner violence (IPV) at wave 1 and compared them on a range of psychosocial outcomes at a 5-year follow-up. This study also examined the rate of divorce or separation during the 5-year interval among women who reported IPV at wave 1 and explored whether certain predictors were related to ending an abusive relationship with an intimate partner during the period. Women with IPV at wave 1, compared to women without IPV, were significantly more likely to experience a greater degree of depressive symptoms and functional impairment and less self-esteem and life satisfaction at the 5-year follow-up. Also, nearly half of the women in an abusive relationship left the relationship within the period. Leaving the abusive relationship was associated with lower individual income and more social support at wave 1. PMID- 16368766 TI - Identification of violence in Turkish health care settings. AB - This study sought to investigate the contributing factors to and frequency of violence against health care workers (HCWs) working in western Turkey. The population is composed of a random sample of 1,209 HCWs from 34 health care workplaces. Written questionnaires were given to HCWs at all sites, where staff were instructed to register all types of violence they had experienced. In all, 49.5% of HCWs reported having experienced verbal, physical, or verbal and physical violence, with this total being made up of 39.6% men and 60.4% women. A larger percentage (69.6%) of general practitioners reported experiencing verbal abuse and physical violence by patients and patients' family members or friends. Younger workers, inexperienced staff, and those in emergency services were more likely to report violence. Violence directed toward HCWs is a common occupational hazard. Public health authorities should plan preventive interventions based on the findings of this study. PMID- 16368767 TI - Bayesian sparse hidden components analysis for transcription regulation networks. AB - MOTIVATION: In systems like Escherichia Coli, the abundance of sequence information, gene expression array studies and small scale experiments allows one to reconstruct the regulatory network and to quantify the effects of transcription factors on gene expression. However, this goal can only be achieved if all information sources are used in concert. RESULTS: Our method integrates literature information, DNA sequences and expression arrays. A set of relevant transcription factors is defined on the basis of literature. Sequence data are used to identify potential target genes and the results are used to define a prior distribution on the topology of the regulatory network. A Bayesian hidden component model for the expression array data allows us to identify which of the potential binding sites are actually used by the regulatory proteins in the studied cell conditions, the strength of their control, and their activation profile in a series of experiments. We apply our methodology to 35 expression studies in E.Coli with convincing results. AVAILABILITY: www.genetics.ucla.edu/labs/sabatti/software.html SUPPLEMENTARY INFORMATION: The supplementary material are available at Bioinformatics online. PMID- 16368768 TI - BioContrasts: extracting and exploiting protein-protein contrastive relations from biomedical literature. AB - MOTIVATION: Contrasts are useful conceptual vehicles for learning processes and exploratory research of the unknown. For example, contrastive information between proteins can reveal what similarities, divergences and relations there are of the two proteins, leading to invaluable insights for better understanding about the proteins. Such contrastive information are found to be reported in the biomedical literature. However, there have been no reported attempts in current biomedical text mining work that systematically extract and present such useful contrastive information from the literature for exploitation. RESULTS: Our BioContrasts system extracts protein-protein contrastive information from MEDLINE abstracts and presents the information to biologists in a web-application for exploitation. Contrastive information are identified in the text abstracts with contrastive negation patterns such as 'A but not B'. A total of 799 169 pairs of contrastive expressions were successfully extracted from 2.5 million MEDLINE abstracts. Using grounding of contrastive protein names to Swiss-Prot entries, we were able to produce 41 471 pieces of contrasts between Swiss-Prot protein entries. These contrastive pieces of information are then presented via a user-friendly interactive web portal that can be exploited for applications such as the refinement of biological pathways. AVAILABILITY: BioContrasts can be accessed at http://biocontrasts.i2r.a-star.edu.sg. It is also mirrored at http://biocontrasts.biopathway.org. SUPPLEMENTARY INFORMATION: Supplementary materials are available at Bioinformatics online. PMID- 16368769 TI - Local RNA base pairing probabilities in large sequences. AB - SUMMARY: The genome-wide search for non-coding RNAs requires efficient methods to compute and compare local secondary structures. Since the exact boundaries of such putative transcripts are typically unknown, arbitrary sequence windows have to be used in practice. Here we present a method for robustly computing the probabilities of local base pairs from long RNA sequences independent of the exact positions of the sequence window. AVAILABILITY: The program RNAplfold is part of the Vienna RNA Package and can be downloaded from http://www.tbi.univie.ac.at/RNA/. PMID- 16368770 TI - Multidimensional local false discovery rate for microarray studies. AB - MOTIVATION: The false discovery rate (fdr) is a key tool for statistical assessment of differential expression (DE) in microarray studies. Overall control of the fdr alone, however, is not sufficient to address the problem of genes with small variance, which generally suffer from a disproportionally high rate of false positives. It is desirable to have an fdr-controlling procedure that automatically accounts for gene variability. METHODS: We generalize the local fdr as a function of multiple statistics, combining a common test statistic for assessing DE with its standard error information. We use a non-parametric mixture model for DE and non-DE genes to describe the observed multi-dimensional statistics, and estimate the distribution for non-DE genes via the permutation method. We demonstrate this fdr2d approach for simulated and real microarray data. RESULTS: The fdr2d allows objective assessment of DE as a function of gene variability. We also show that the fdr2d performs better than commonly used modified test statistics. AVAILABILITY: An R-package OCplus containing functions for computing fdr2d() and other operating characteristics of microarray data is available at http://www.meb.ki.se/~yudpaw. PMID- 16368771 TI - FLOSS: flexible ordered subset analysis for linkage mapping of complex traits. AB - The FLOSS software package is a flexible framework for ordered subset analysis. FLOSS is specifically designed for use with the Merlin linkage analysis package, but FLOSS can be used with any linkage analysis software package that reports NPL Z-scores for each locus and family. When FLOSS is used with the Merlin linkage analysis package, one can use either non-parametric Z-scores or Kong and Cox linear allele sharing model LOD scores. Monte Carlo P-values are calculated using a permutation test with an efficient Besag-Clifford sequential stopping rule. FLOSS also has a flexible tool for assigning family covariate scores from Merlin input files. FLOSS includes user documentation and is written in Java for easy portability. The FLOSS source code is documented and designed to be extensible. PMID- 16368772 TI - Accurate prediction of HIV-1 drug response from the reverse transcriptase and protease amino acid sequences using sparse models created by convex optimization. AB - MOTIVATION: Genotype-phenotype modeling problems are often overcomplete, or ill posed, since the number of potential predictors-genes, proteins, mutations and their interactions-is large relative to the number of measured outcomes. Such datasets can still be used to train sparse parameter models that generalize accurately, by exerting a principle similar to Occam's Razor: When many possible theories can explain the observations, the most simple is most likely to be correct. We apply this philosophy to modeling the drug response of Type-1 Human Immunodeficiency Virus (HIV-1). Owing to the decreasing expense of genetic sequencing relative to in vitro phenotype testing, a statistical model that reliably predicts viral drug response from genetic data is an important tool in the selection of antiretroviral therapy (ART). The optimization techniques described will have application to many genotype-phenotype modeling problems for the purpose of enhancing clinical decisions. RESULTS: We describe two regression techniques for predicting viral phenotype in response to ART from genetic sequence data. Both techniques employ convex optimization for the continuous subset selection of a sparse set of model parameters. The first technique, the least absolute shrinkage and selection operator, uses the l(1) norm loss function to create a sparse linear model; the second, the support vector machine with radial basis kernel functions, uses the epsilon-insensitive loss function to create a sparse non-linear model. The techniques are applied to predict the response of the HIV-1 virus to 10 reverse transcriptase inhibitor and 7 protease inhibitor drugs. The genetic data are derived from the HIV coding sequences for the reverse transcriptase and protease enzymes. When tested by cross-validation with actual laboratory measurements, these models predict drug response phenotype more accurately than models previously discussed in the literature, and other canonical techniques described here. Key features of the methods that enable this performance are the tendency to generate simple models where many of the parameters are zero, and the convexity of the cost function, which assures that we can find model parameters to globally minimize the cost function for a particular training dataset. AVAILABILITY: Results, tables and figures are available at ftp://ftp.genesecurity.net. SUPPLEMENTARY INFORMATION: An Appendix to accompany this article is available at Bioinformatics online. PMID- 16368773 TI - MiGenes: a searchable interspecies database of mitochondrial proteins curated using gene ontology annotation. AB - MOTIVATION: There has been an explosion of interest in the role of mitochondria in programmed cell death and other fundamental pathological processes underlying the development of human diseases. Nevertheless, the inventory of mitochondrial proteins encoded in the nuclear genome remains incomplete, providing an impediment to mitochondrial research at the interface with systems biology. We created the MiGenes database to further define the scope of the mitochondrial proteome in humans and model organisms including mice, rats, flies and worms as well as budding and fission yeasts. MiGenes is intended to stimulate mitochondrial research using model organisms. SUMMARY: MiGenes is a large-scale relational database that is automatically updated to keep pace with advances in mitochondrial proteomics and is curated to assure that the designation of proteins as mitochondrial reflects gene ontology (GO) annotations supported by high-quality evidence codes. A set of postulates is proposed to help define which proteins are authentic components of mitochondria. MiGenes incorporates >1160 new GO annotations to human, mouse and rat protein records, 370 of which represent the first GO annotation reflecting a mitochondrial localization. MiGenes employs a flexible search interface that permits batchwise accession number searches to support high-throughput proteomic studies. A web interface is provided to permit members of the mitochondrial research community to suggest modifications in protein annotations or mitochondrial status. PMID- 16368774 TI - Multivariate survival analysis for case-control family data. AB - Multivariate survival data arise from case-control family studies in which the ages at disease onset for family members may be correlated. In this paper, we consider a multivariate survival model with the marginal hazard function following the proportional hazards model. We use a frailty-based approach in the spirit of Glidden and Self (1999) to account for the correlation of ages at onset among family members. Specifically, we first estimate the baseline hazard function nonparametrically by the innovation theorem, and then obtain maximum pseudolikelihood estimators for the regression and correlation parameters plugging in the baseline hazard function estimator. We establish a connection with a previously proposed generalized estimating equation-based approach. Simulation studies and an analysis of case-control family data of breast cancer illustrate the methodology's practical utility. PMID- 16368775 TI - Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. What is the role of genome duplication in the evolution of complexity and diversity? AB - Gene and genome duplications provide a source of genetic material for mutation, drift, and selection to act upon, making new evolutionary opportunities possible. As a result, many have argued that genome duplication is a dominant factor in the evolution of complexity and diversity. However, a clear correlation between a genome duplication event and increased complexity and diversity is not apparent, and there are inconsistencies in the patterns of diversity invoked to support this claim. Interestingly, several estimates of genome duplication events in vertebrates are preceded by multiple extinct lineages, resulting in preduplication gaps in extant taxa. Here we argue that gen(om)e duplication could contribute to reduced risk of extinction via functional redundancy, mutational robustness, increased rates of evolution, and adaptation. The timeline for these processes to unfold would not predict immediate increases in species diversity after the duplication event. Rather, reduced probabilities of extinction would predict a latent period between a genome duplication and its effect on species diversity or complexity. In this paper, we will develop the idea that genome duplication could contribute to species diversity through reduced probability of extinction. PMID- 16368776 TI - Diversity and evolution of the thyroglobulin type-1 domain superfamily. AB - Multidomain proteins are gaining increasing consideration for their puzzling, flexible utilization in nature. The presence of the characteristic thyroglobulin type-1 (Tg1) domain as a protein module in a variety of multicellular organisms suggests pivotal roles for this building block. To gain insight into the evolution of Tg1 domains, we performed searches of protein, expressed sequence tag, and genome databases. Tg1 domains were found to be Metazoa specific, and we retrieved a total of 170 Tg1 domain-containing protein sequences. Their architectures revealed a wide taxonomic distribution of proteins containing Tg1 domains followed or preceded by secreted protein, acidic, rich in cysteines (SPARC)-type extracellular calcium-binding domains. Other proteins contained lineage-specific domain combinations of peptidase inhibitory modules or domains with different biological functions. Phylogenetic analysis showed that Tg1 domains are highly conserved within protein structures, whereas insertion into novel proteins is followed by rapid diversification. Seven different basic types of protein architecture containing the Tg1 domain were identified in vertebrates. We examined the evolution of these protein groups by combining Tg1 domain phylogeny with additional analyses based on other characteristic domains. Testicans and secreted modular calcium binding protein (SMOCs) evolved from invertebrate homologs by introduction of vertebrate-specific domains, nidogen evolved by insertion of a Tg1 domain into a preexisting architecture, and the remaining four have unique architectures. Thyroglobulin, Trops, and the major histocompatibility complex class II-associated invariant chain are vertebrate specific, while an insulin-like growth factor-binding protein and nidogen were also identified in urochordates. Among vertebrates, we observed differences in protein repertoires, which result from gene duplication and domain duplication. Members of five groups have been characterized at the molecular level. All exhibit subtle differences in their specificities and function either as peptidase inhibitors (thyropins), substrates, or both. As far as the sequence is concerned, only a few conserved residues were identified. In combination with structural data, our analysis shows that the Tg1 domain fold is highly adaptive and comprises a relatively well-conserved core surrounded by highly variable loops that account for its multipurpose function in the animal kingdom. PMID- 16368777 TI - Alternatively and constitutively spliced exons are subject to different evolutionary forces. AB - There has been a controversy on whether alternatively spliced exons (ASEs) evolve faster than constitutively spliced exons (CSEs). Although it has been noted that ASEs are subject to weaker selective constraints than CSEs, so they evolve faster, there have also been studies that indicated slower evolution in ASEs than in CSEs. In this study, we retrieve more than 5,000 human-mouse orthologous exons and calculate the synonymous (KS) and nonsynonymous (KA) substitution rates in these exons. Our results show that ASEs have higher KA values and higher KA/KS ratios than CSEs, indicating faster amino acid-level evolution in ASEs. The faster evolution may be in part due to weaker selective constraints. It is also possible that the faster rate is in part due to faster functional evolution in ASEs. On the other hand, the majority of ASEs have lower KS values than CSEs. With reference to the substitution rate in introns, we show that the KS values in ASEs are close to the neutral substitution rate, whereas the synonymous substitution rate in CSEs has likely been accelerated. The elevated synonymous rate in CSEs is not related to CpG dinucleotides or low-complexity regions of protein but may be weakly related to codon usage bias. The overall trends of higher KA and lower KS in ASEs than in CSEs are also observed in human-rat and mouse-rat comparisons. Therefore, our observations hold for mammals of different molecular clocks. PMID- 16368778 TI - Comparative analysis of bacterial-origin genes for plant mitochondrial ribosomal proteins. AB - Mitochondrial ribosomes contain bacterial-type proteins reflecting their endosymbiotic heritage, and a subset of these genes is retained within the mitochondrion in land plants. Variation in gene location is observed, however, because migration to the nucleus is still an ongoing evolutionary process in plants. To gain insights into adaptation events related to successful gene transfer, we have compiled data for bacterial-origin mitochondrial-type ribosomal protein genes from the completely sequenced Arabidopsis and rice genomes. Approximately 75% of such nuclear-located genes encode amino-terminal extensions relative to their Escherichia coli counterparts, and of that set, only about 30% have introns at (or near) the junction in support of an exon shuffling-type recruitment of upstream expression/targeting signals. We find that genes that were transferred to the nucleus early in eukaryotic evolution have, on average, about twofold higher density of introns within the core ribosomal protein sequences than do those that moved to the nucleus more recently. About 20% of such introns are at positions identical to those in human orthologs, consistent with their ancestral presence. Plant mitochondrial-type ribosomal protein genes have dispersed chromosomal locations in the nucleus, and about 20% of them are present in multiple unlinked copies. This study provides new insights into the evolutionary history of endosymbiotic bacterial-type genes that have been transferred from the mitochondrion to the nucleus. PMID- 16368779 TI - The contribution of LTR retrotransposon sequences to gene evolution in Mus musculus. AB - Approximately 1.5% of mouse genes (Mus musculus) contain long terminal repeat retrotransposon sequences (LRS). Consistent with earlier findings in Caenorhabditis elegans, Drosophila melanogaster, and Homo sapiens, LRS are more likely to be associated with newly evolved genes. Evidence is presented that LRS are often recruited as novel exons or as spliced additions to existing exons. These novel gene configurations may be expressed initially as alternative transcripts providing an opportunity for the evolution of new gene function. PMID- 16368780 TI - Impaired transforming growth factor beta signalling in Barrett's carcinogenesis due to frequent SMAD4 inactivation. AB - BACKGROUND AND AIMS: Transforming growth factor beta (TGF-beta) is frequently involved in gastrointestinal carcinogenesis although its contribution to oesophageal adenocarcinoma (AC) and its precursor Barrett's oesophageal epithelium (BE) metaplasia are unclear. METHODS: Expression of TGF-beta signalling components was assessed by reverse transcription-polymerase chain reaction (PCR), western blot, and immunohistochemistry in oesophageal endoscopic biopsies and cell lines. Genomic alterations in SMAD4 were characterised by fluorescence in situ hybridisation, methylation specific PCR, and sequencing. Functional integrity of TGF-beta signalling was assessed by characterisation of p21 and proliferation status. Smad4 negative BIC-1 cells were transiently transfected with smad4 and TGF-beta responsiveness evaluated. RESULTS: smad4 mRNA expression was progressively reduced in the metaplasia-dysplasia-adenocarcinoma sequence (p<0.01). A quarter of AC samples displayed an abnormal Smad4 protein isoform, with no corresponding changes in gene sequence or organisation. Methylation of smad4 has not been described previously but we found promoter methylation in 70% of primary AC samples. In 6/8 oesophageal cell lines, chromosomal rearrangements affected the smad4 locus. Lack of smad4 expression in BIC-1 cells occurred secondary to loss of one copy and extensive deletion of the second allele's promoter region. TGF-beta dependent induction of p21 and downregulation of minichromosome maintenance protein 2 was lost in >80% of BE and AC. TGF-beta failed to inhibit proliferation in 5/8 oesophageal cell lines. In BIC-1, the antiproliferative response was restored following transient transfection of smad4 cDNA. CONCLUSIONS: In BE carcinogenesis, downregulation of Smad4 occurs due to several different mechanisms, including methylation, deletion, and protein modification. Frequent alterations in TGF-beta signalling lead to a functionally significant impairment of TGF-beta mediated growth suppression. PMID- 16368781 TI - CXCR3 and alphaEbeta7 integrin identify a subset of CD8+ mature thymocytes that share phenotypic and functional properties with CD8+ gut intraepithelial lymphocytes. AB - BACKGROUND: We previously demonstrated the existence of two distinct subsets of T cell receptor (TCR)alphabeta+CD8alphabeta+ single positive (SP) cells in human postnatal thymus which express the chemokine receptor CCR7 or CXCR3 and migrate in vitro in response to their specific ligands. AIM: To investigate whether these two CD8+ thymocyte subsets had distinct peripheral colonisation. METHODS: TCRalphabeta+CD8+ SP cells were obtained from normal postnatal thymus, mesenteric lymph node (LNs), small bowel, and peripheral blood (PB) specimens. Cells were then evaluated for expression of surface molecules, cytolytic potential, telomere length, and profile of cytokine production. RESULTS: CD8+CCR7+CXCR3- thymocytes exhibited CD62L, in common with those which localise to LNs. In contrast, CD8+CCR7-CXCR3+ thymocytes lacked CD62L but exhibited CD103, similar to intraepithelial lymphocytes (IELs) present in the gut mucosa where the CXCR3 ligand, CXCL10, and the CD103 ligand, E-cadherin, are highly and consistently expressed. In addition, thymocytes and gut CD8+CXCR3+CD103+ cells showed comparable telomere length, which was higher than that of PB CXCR3+CD8+ T cells. However, both of these populations contained perforin and granzyme A, and displayed the ability to produce interferon gamma and interleukin 2. Of note, CXCR3 deficient, in comparison with wild-type C57Black/6, mice showed decreased proportions of CD3+CD8alphabeta+ and increased proportions of CD3+CD8alphaalpha+ lymphocytes at gut level. Moreover, adoptive transfer of CD3+CD8alphabeta+ thymocytes from wild-type into CXCR3 deficient mice resulted in a significant increase in CD3+CD8alphabeta+ T cells in the gut mucosa but not in other tissues. CONCLUSIONS: The results of this study demonstrate the existence of a previously unrecognised subset of TCRalphabeta+CD8alphabeta+ SP CXCR3+CD103+ thymocytes which share phenotypic and functional features with CD8+ IELs, thus suggesting the possibility of their direct colonisation of the gut mucosa. PMID- 16368782 TI - DHT and testosterone, but not DHEA or E2, differentially modulate IGF-I, IGFBP-2, and IGFBP-3 in human prostatic stromal cells. AB - Prostate cancer is one of the four most common cancers in the United States, affecting one of six men. Increased serum levels of androgens and IGF-I are associated with an augmented risk of prostate cancer. Dihydrotestosterone (DHT) and testosterone (T) stimulate prostate cancer cell growth, development, and function, whereas the effects of DHT and T in prostate stromal cells, and of dehydroepiandrosterone (DHEA) in prostate cancer or stromal cells, are uncertain. We investigated the actions of DHT, T, DHEA, and estradiol (E2) on insulin-like growth factor (IGF)-I, IGF-II, IGF-I receptor (R), IGF-binding protein (IGFBP)-2, IGFBP-3, and IGFBP-5 in primary cultures of human prostatic stromal cells by assessing cell proliferation, mRNA expression, and protein secretion by MTT growth assay, quantitative real-time PCR, and ELISA, respectively. DHT and T each increased IGF-I (7-fold) and decreased IGFBP-3 (2-fold) mRNA expression and protein secretion in a dose- and time-dependent manner and increased IGFBP-2 (2 fold) mRNA in a dose- and time-dependent manner. DHEA and E2 did not significantly alter these measures. Flutamide abolished the DHT-modulated increases in IGF-I and IGFBP-2, suggesting that the influences of DHT and T on these measures were androgen receptor mediated. None of the four steroids significantly affected IGF-IR, IGF-II, or IGFBP-5 mRNA levels or stromal cell proliferation. The effects of DHT on IGF-I, IGFBP-2, and IGFBP-3 were more pronounced in stromal cultures that did not express desmin. These data suggest that DHT and T promote prostate growth partly via modulation of the stromal cell IGF axis, with potential paracrine effects on prostate epithelial cells. PMID- 16368783 TI - Attenuated corticosterone response to chronic ACTH stimulation in hepatic lipase deficient mice: evidence for a role for hepatic lipase in adrenal physiology. AB - Hepatic lipase (HL), a liver-expressed lipolytic enzyme, hydrolyzes triglycerides and phospholipids in lipoproteins and promotes cholesterol delivery through receptor-mediated whole particle and selective cholesterol uptake. HL activity also occurs in the adrenal glands, which utilize lipoprotein cholesterol to synthesize glucocorticoids in response to pituitary ACTH. It is likely that the role of adrenal HL is to facilitate delivery of exogenous cholesterol for glucocorticoid synthesis. On this basis, we hypothesized that HL deficiency would blunt the glucocorticoid response to ACTH. Furthermore, because exogenous cholesterol also is derived from the LDL receptor (LDLR) pathway, we hypothesized that LDLR deficiency would blunt the response to ACTH. To test these hypotheses, we compared the corticosterone response to eight daily ACTH injections in HL deficient (hl-/-), LDLR-deficient (Ldlr-/-), and HL- and LDLR-doubly deficient (Ldlr-/- hl-/-) mice with that in wild-type (WT) mice. Plasma corticosterone levels were measured on days 2, 5, and 8. Differences in plasma corticosterone levels between genotypes were analyzed by Kruskal-Wallis one-way ANOVA on ranks and pairwise multiple comparisons by Dunn's test. Our results demonstrate a trend toward reductions in plasma corticosterone levels on day 2 and significant reductions on day 5 and day 8 in the knockout models. Thus, on day 5, plasma corticosterone levels were reduced by 57, 70, and 73% (all P < 0.05) and on day 8 by 76, 59, and 63% (all P < 0.05) in hl-/-, Ldlr-/-, and Ldlr-/- hl-/- mice, respectively. These results demonstrate that HL deficiency, like LDLR deficiency, blunts the adrenal response to chronic ACTH stimulation and suggest a novel role for HL in adrenal physiology. PMID- 16368784 TI - Molecular mechanism of 1,25-dihydroxyvitamin D3 inhibition of adipogenesis in 3T3 L1 cells. AB - We have investigated the molecular mechanism whereby 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] inhibits adipogenesis in vitro. 1,25(OH)2D3 blocks 3T3-L1 cell differentiation into adipocytes in a dose-dependent manner; however, the inhibition is ineffective 24-48 h after the differentiation is initiated, suggesting that 1,25(OH)2D3 inhibits only the early events of the adipogenic program. Treatment of 3T3-L1 cells with 1,25(OH)2D3 does not block the mitotic clonal expansion or C/EBPbeta induction; rather, 1,25(OH)2D3 blocks the expression of C/EBPalpha, peroxisome proliferator-activated receptor-gamma (PPARgamma), sterol regulatory element-binding protein-1, and other downstream adipocyte markers. The inhibition by 1,25(OH)2D3 is reversible, since removal of 1,25(OH)2D3 from the medium restores the adipogenic process with only a temporal delay. Interestingly, although the vitamin D receptor (VDR) protein is barely detectable in 3T3-L1 preadipocytes, its levels are dramatically increased during the early phase of adipogenesis, peaking at 4-8 h and subsiding afterward throughout the rest of the differentiation program; 1,25(OH)2D3 treatment appears to stabilize the VDR protein levels. Consistently, adenovirus-mediated overexpression of human (h) VDR in 3T3-L1 cells completely blocks the adipogenic program, confirming that VDR is inhibitory. Inhibition of adipocyte differentiation by 1,25(OH)2D3 is ameliorated by troglitazone, a specific PPARgamma antagonist; conversely, hVDR partially suppresses the transacting activity of PPARgamma but not of C/EBPbeta or C/EBPalpha. Moreover, 1,25(OH)2D3 markedly suppresses C/EBPalpha and PPARgamma mRNA levels in mouse epididymal fat tissue culture. Taken together, these data indicate that the blockade of 3T3-L1 cell differentiation by 1,25(OH)2D3 occurs at the postclonal expansion stages and involves direct suppression of C/EBPalpha and PPARgamma upregulation, antagonization of PPARgamma activity, and stabilization of the inhibitory VDR protein. PMID- 16368785 TI - Imaging pancreatic beta-cells in the intact pancreas. AB - We have developed a method to visualize fluorescent protein-labeled beta-cells in the intact pancreas through combined reflection and confocal imaging. This method provides a 3-D view of the beta-cells in situ. Imaging of the pancreas from mouse insulin I promoter (MIP)-green (GFP) and red fluorescent protein (RFP) transgenic mice shows that islets, beta-cell clusters, and single beta-cells are not evenly distributed but are aligned along the large blood vessels. We also observe the solitary beta-cells in both fetal and adult mice and along the pancreatic and common bile ducts. We have imaged the developing endocrine cells in the embryos using neurogenin-3 (Ngn3)-GFP mice crossed with MIP-RFP mice. The dual-color coded pancreas from embryos (E15.5) shows a large number of green Ngn3-expressing proendocrine cells with a smaller number of red beta-cells. The imaging technique that we have developed, coupled with the transgenic mice in which beta-cells and beta-cell progenitors are labeled with different fluorescent proteins, will be useful for studying pancreatic development and function in normal and disease states. PMID- 16368786 TI - Novel application of the "doubly labeled" water method: measuring CO2 production and the tissue-specific dynamics of lipid and protein in vivo. AB - The partitioning of whole body carbon flux between fat and lean compartments affects body composition. We hypothesized that it is possible to simultaneously determine whole body carbon (energy) balance and the dynamics of lipids and proteins in specific tissues in vivo. Growing C57BL/6J mice fed a high-fat low carbohydrate diet were injected with a bolus of "doubly labeled" water (i.e., (2)H2O and H2(18)O). The rate of CO2 production was determined from the difference between the elimination rates of 2H and 18O from body water. The rates of synthesis and degradation of triglycerides extracted from epididymal fat pads and of proteins extracted from heart muscle were determined by mathematically modeling the 2H labeling of triglyceride-bound glycerol and protein-bound alanine, respectively. We found that mice were in positive carbon balance (approximately 20% retention per day) and accumulated lipid in epididymal fat pads (approximately 9 micromol triglyceride accumulated per day). This is consistent with the fact that mice were studied during a period of growth. Modeling the 2H labeling of triglycerides revealed a substantial rate of lipid breakdown during this anabolic state (equivalent to approximately 25% of the newly synthesized triglyceride). We found equal rates of protein synthesis and breakdown in heart muscle (approximately 10% of the pool per day), consistent with the fact that the heart muscle mass did not change. In total, these findings demonstrate a novel application of the doubly labeled water method. Utilization of this approach, especially in unique rodent models, should facilitate studies aimed at quantifying the efficacy of interventions that modulate whole body carbon balance and lipid flux while in parallel determining their impact on (cardiac) muscle protein turnover. Last, the simplicity of administering doubly labeled water and collecting samples allows this method to be used in virtually any laboratory setting. PMID- 16368787 TI - Characterization of novel Na+-dependent nucleobase transport systems at the blood testis barrier. AB - In the testis, nucleosides and nucleobases are important substrates of the salvage pathway for nucleotide biosynthesis, and one of the roles of Sertoli cells is to provide nutrients and metabolic precursors to spermatogenic cells located within the blood-testis barrier (BTB). We have already shown that concentrative and equilibrative nucleoside transporters are expressed and are functional in primary-cultured rat Sertoli cells as a BTB model, but little is known about nucleobase transport at the BTB or about the genes encoding specific nucleobase transporters in mammalian cells. In the present study, we examined the uptake of purine ([3H]guanine) and pyrimidine ([3H]uracil) nucleobases by primary cultured rat Sertoli cells. The uptake of both nucleobases was time and concentration dependent. Kinetic analysis showed the involvement of three different transport systems in guanine uptake. In contrast, uracil uptake was mediated by a single Na+-dependent high-affinity transport system. Guanine uptake was inhibited by other purine nucleobases but not by pyrimidine nucleobases, whereas uracil uptake was inhibited only by pyrimidine nucleobases. In conclusion, it was suggested that there might be purine- or pyrimidine-selective nucleobase transporters in rat Sertoli cells. PMID- 16368788 TI - Indispensable role of mitochondrial UCP1 for antiobesity effect of beta3 adrenergic stimulation. AB - Mitochondrial uncoupling protein-1 (UCP1) has been thought to be a key molecule for thermogenesis during cold exposure and spontaneous hyperphagia and thereby in the autonomic regulation of energy expenditure and adiposity. However, UCP1 knockout (KO) mice were reported to be cold intolerant but unexpectedly did not get obese even after hyperphagia, implying that UCP1 may not be involved in the regulation of adiposity. Treatment of obese animals with beta3-adrenergic agonists is known to increase lipid mobilization, induce UCP1, and, finally, reduce body fat content. To obtain direct evidence for the role of UCP1 in the anti-obesity effect of beta3-adrenergic stimulation, in the present study, UCP1 KO and wild-type (WT) mice were fed on cafeteria diets for 8 wk and then given a beta3-adrenergic agonist, CL-316,243 (CL), or saline for 2 wk. A single injection of CL increased whole body oxygen consumption and brown fat temperature in WT mice but not in KO mice, and it elicited almost the same plasma free fatty acid response in WT and KO mice. WT and KO mice increased similarly their body and white fat pad weights on cafeteria diets compared with those on laboratory chow. Daily treatment with CL resulted in a marked reduction of white fat pad weight and the size of adipocytes in WT mice, but not in KO mice. Compared with WT mice, KO mice expressed increased levels of UCP2 in brown fat but decreased levels in white fat and comparable levels of UCP3. It was concluded that the anti-obesity effect of beta3-adrenergic stimulation is largely attributable to UCP1, but less to UCP2 and UCP3, and thereby to UCP1-dependent degradation of fatty acids released from white adipose tissue. PMID- 16368789 TI - Bombesin-like peptides: modulators of inflammation in acute lung injury? PMID- 16368790 TI - Lymphangioleiomyomatosis: a national registry for a rare disease. PMID- 16368791 TI - Muscle wasting in chronic obstructive pulmonary disease: to bother and to measure! PMID- 16368792 TI - PTEN as a new agent in the fight against fibrogenesis. PMID- 16368793 TI - Body mass, fat-free body mass, and prognosis in patients with chronic obstructive pulmonary disease from a random population sample: findings from the Copenhagen City Heart Study. AB - RATIONALE: Low body mass index (BMI) is a marker of poor prognosis in chronic obstructive pulmonary disease (COPD). In the general population, the harmful effect of low BMI is due to the deleterious effects of a low fat-free mass index (FFMI; fat-free mass/weight(2)). OBJECTIVES: We explored distribution of low FFMI and its association with prognosis in a population-based cohort of patients with COPD. METHODS: We used data on 1,898 patients with COPD identified in a population-based epidemiologic study in Copenhagen. FFM was measured using bioelectrical impedance analysis. Patients were followed up for a mean of 7 yr and the association between BMI and FFMI and mortality was examined taking age, sex, smoking, and lung function into account. MAIN RESULTS: The mean FFMI was 16.0 kg/m(2) for women and 18.7 kg/m(2) for men. Among subjects with normal BMI, 26.1% had an FFMI lower than the lowest 10th percentile of the general population. BMI and FFMI were significant predictors of mortality, independent of relevant covariates. Being in the lowest 10th percentile of the general population for FFMI was associated with a hazard ratio of 1.5 (95% confidence interval, 1.2-1.8) for overall mortality and 2.4 (1.4-4.0) for COPD-related mortality. FFMI was also a predictor of overall mortality when analyses were restricted to subjects with normal BMI. CONCLUSIONS: FFMI provides information in addition to BMI and assessment of FFM should be considered in the routine assessment of COPD. PMID- 16368794 TI - Use of commercial interferon-gamma assays in immunocompromised patients for tuberculosis diagnosis. PMID- 16368795 TI - Adult hospital and ventilator-associated pneumonia guidelines: eminence- rather than evidence-based. PMID- 16368796 TI - Reinfection with Mycobacterium tuberculosis. PMID- 16368797 TI - Duration of dialysis is a significant predictor of prolonged postoperative mechanical ventilation in dialysis-dependent patients undergoing cardiac surgery. AB - Prolonged mechanical ventilation is reported to correlate with increased risk of mortality after cardiac surgery. We designed the present study to determine the preoperative and intraoperative risk factors that could predict postoperative prolonged mechanical ventilation in dialysis-dependent patients undergoing cardiac surgery with cardiopulmonary bypass. Forty-four dialysis-dependent patients were divided into two groups; patients of group E were tracheally extubated within 24 h after admission to the intensive care unit postoperatively (n = 19) and patients of group L (n = 25) required more than 24 h of mechanical ventilation. All patients received hemofiltration during cardiopulmonary bypass and continuous veno-venous hemodialysis postoperatively. A multiple logistic regression analysis showed that duration of dialysis (>10 yr) and duration of surgery (>8 h) were independent risk factors of prolonged mechanical ventilation (>24 h). On admission to the intensive care unit, Pao2/Fio2 of group L was significantly lower than that of group E (294 +/- 135 versus 415 +/- 99 mm Hg) and the circulatory status of group L was worse than that of group E. The median (interquartile range) duration of intensive care unit stay in group E was 3 (3.00) days, which was significantly shorter than that of group L (5 [2.75] days). It is possible that longer surgery increases the likelihood of cardiac dysfunction and poor oxygenation in patients with a long history of dialysis. PMID- 16368798 TI - A meta-analytic comparison of preoperative stress echocardiography and nuclear scintigraphy imaging. AB - In this meta-analysis we compared thallium imaging (TI) and stress echocardiography (SE) in patients at risk for myocardial infarction (MI) scheduled for elective noncardiac surgery. Two searches of published articles were used to identify relevant articles. We included all studies that stated the criteria for a positive test and detailed the frequency of postoperative MI and in-hospital death. Data were abstracted by two authors and captured preoperative patient characteristics, study design, blinding, and outcome adjudication. We defined a positive test as a test with a reversible defect and, where possible, quantified the size of the defects in each study. MI and/or death were the only postoperative outcomes of interest. We calculated the sensitivity, specificity, and likelihood ratio (LR) and, where possible, the Receiver Operating Characteristic (ROC) curve of a cardiac event in each study. The LR and ROC were combined by meta-analyses using the random effects model. Heterogeneity was assessed using the I2 test. The search revealed 68 studies of 10,049 patients. There were 25 SE studies and 50 TI studies. There were 7 studies with a direct comparison of the two methodologies. The quality of studies differed; routine screening for MI was used more frequently in SE studies (47.8% versus 21.2%; P = 0.008) and screening dictated treatment more often after TI (72.1%) than after SE (46.3%) (P = 0.027). The LR for SE was more indicative of a postoperative cardiac event than TI (LR, 4.09; 95% CI, 3.21-6.56 versus 1.83; 1.59-2.10; P = 0.001). This difference was attributable to fewer false-negative SEs. There was no difference in the cumulative ROC curves from qualitative studies (SE, 0.80; 95% CI, 0.76-.84 versus TI, 0.75; 95% CI, 0.70-081). Again, the LR for a negative SE was less (0.23; 95% CI, 0.17-0.32 versus 0.44; 95% CI, 0.36-0.54). A moderate-to large defect, seen in 14% of patients, by either method predicts a postoperative cardiac event (LR, 8.35; 95% CI, 5.6-12.45). This meta-analysis possesses the statistical power to demonstrate that SE has better negative predicative characteristics than TI. A moderate-to-large perfusion defect by either SE or TI predicts postoperative MI and death. We conclude the SE is superior to TI in predicting postoperative cardiac events. PMID- 16368799 TI - Small-dose epoprostenol decreases systemic oxygen consumption and splanchnic oxygen extraction during normothermic cardiopulmonary bypass. AB - Normothermic, nonpulsatile cardiopulmonary bypass (CPB) impairs systemic and splanchnic oxygen transport and increases gastrointestinal permeability. It is an important therapeutic goal to avoid splanchnic dysoxia during CPB. Small-dose prostacyclin therapy improves splanchnic oxygen transport and microcirculation in septic patients. In this study, we sought to determine if during cardiac surgery, the prostacyclin analog epoprostenol improves the balance of systemic and splanchnic oxygen transport. Eighteen patients undergoing cardiac valve replacement were randomized to receive either epoprostenol (3 ng x kg(-1) x min( 1)) or placebo during, and for 1 hour after, surgery. Systemic and splanchnic oxygen delivery, consumption, and extraction and arterial, mixed venous, and hepato-venous lactate concentrations were measured before, during, and after CPB. Gastrointestinal permeability was measured 1 day before and 1 day after surgery using the triple sugar permeability test. During CPB, the epoprostenol group had decreased systemic oxygen consumption and splanchnic oxygen extraction (P = 0.024). These effects were not present 1 hour after the end of epoprostenol infusion. The study was not adequately powered to determine whether epoprostenol altered the trend towards increased lactate metabolism and increased postoperative gastrointestinal permeability, nor could we demonstrate any differences between groups in clinically relevant end-points. In conclusion, these findings suggest that during normothermic CPB, small-dose epoprostenol therapy may reduce systemic oxygen consumption and splanchnic oxygen extraction. PMID- 16368800 TI - Symmetry aortic connector devices and acute renal injury: a comparison of renal dysfunction after three different aortocoronary bypass surgery techniques. AB - Although the pathogenesis of acute renal injury after cardiac surgery is multifactorial, atherosclerosis of the ascending aorta and embolic burden are strong independent predictors. Use of the Symmetry aortic connector device (ACD) for proximal anastomosis of coronary grafts may reduce ascending aortic atheroembolism. Therefore, we tested the hypothesis that off-pump coronary artery bypass (OPCAB) surgery performed using an ACD is associated with less postoperative renal dysfunction compared with conventional OPCAB or on-pump coronary artery bypass graft (CABG) surgery. Three-thousand-three-hundred consecutive patients undergoing non-emergent aortocoronary bypass surgery were retrospectively divided into three groups by surgical procedure; Group A: OPCAB with ACD (n = 124), Group B: standard OPCAB (n = 313), Group C: on-pump CABG (n = 2863). Postoperative peak fractional change in creatinine compared with baseline was used as a measure of renal outcome. Multivariable analysis did not identify ACD use as an independent predictor of postoperative peak fractional change in creatinine (P = 0.71), although the relationships of several known renal risk factors with postoperative peak fractional change in creatinine were confirmed. We could not find evidence that OPCAB surgery using ACDs reduces acute renal injury compared with standard OPCAB or CABG surgery. PMID- 16368801 TI - Cardiomyopathic etiology and SERCA2a reverse remodeling during mechanical support of the failing human heart. AB - Many hearts in end-stage, chronic failure (CHF) retain the capacity to reverse abnormal expression of genes regulating myocyte calcium cycling when supported with a left ventricular assist device (LVAD). In the present study, we determined whether LVAD-induced upregulation of the gene encoding for the key calcium cycling protein sarcoplasmic endoreticular calcium adenosine triphosphatase subtype 2a (SERCA2a) is influenced by the nature of underlying disease broadly characterized as ischemic (ICM) or idiopathic dilated (DCM) cardiomyopathy. Data from Northern blot analysis of SERCA2a messenger (m)RNA within 84 heart samples (50 CHF [23 DCM and 27 ICM] and 34 CHF+LVAD [18 DCM and 16 ICM]) were used for characterizing gene expression. In addition, measurements of the force-frequency relationship (FFR), a reflection of in vivo SERCA2a function, were obtained in myocardial trabeculae isolated from 75 hearts (51 CHF [29 DCM and 22 ICM] and 24 CHF+LVAD [10 DCM and 14 ICM]). SERCA2a mRNA demonstrated upregulation after LVAD that was not influenced by ICM or DCM. However, only in DCM hearts was the proportion of trabeculae exhibiting a normal FFR increased after LVAD. Thus, although upregulated SERCA2a gene expression after LVAD support is independent of myopathic origin, normalization of myocardial FFR, an index of SERCA2a function, is not. These data provide new insight into the process of cardiac "reverse molecular remodeling," and underscore potential differences in the impact of disease processes on posttranscriptional events. PMID- 16368802 TI - Effects of sevoflurane anesthesia on carotid-cardiac baroreflex responses in humans. AB - Sevoflurane depresses cardio-vagal baroreflex gain (ability of vagally mediated R R interval response to arterial blood pressure change). We examined the effects of sevoflurane anesthesia on maximum buffering capacity of vagally mediated hemodynamic control (baroreflex range) by examining the entire stimulus-response baroreflex relation. Electrocardiogram and invasive arterial blood pressure were monitored in 11 healthy volunteers. Carotid-cardiac baroreflex responses were elicited by increasing neck chamber pressure (external pressure applied over the bilateral carotid sinuses) to 40 mm Hg for 5 heartbeats followed by decreasing chamber pressure by successive 15-mm Hg R-wave triggered decrements to -65 mm Hg during held expiration. R-R intervals were plotted as functions of preceding carotid distending pressure. Range, maximum gain, and operational point (relative position of the resting set point within the entire baroreflex response curve) were determined at conscious baseline, during 2% (end-tidal) sevoflurane anesthesia, without and with phenylephrine infusion to maintain conscious arterial blood pressure, and at 30, 60, 120, and 180 min after emergence from anesthesia. Sevoflurane anesthesia significantly depressed maximum gain (from 3.84 +/- 0.99 to 1.04 +/- 0.40 ms/mm Hg [mean +/- sd]; P < 0.001) and range (from 207 +/- 43 to 52 +/- 19 ms; P < 0.001) of the reflex relation, both of which recovered at 120 and 180 min after emergence. Phenylephrine infusion only partially restored these variables. The operational point was unchanged throughout the study. Our results indicate that maximum cardio-vagal compensatory response to buffer hemodynamic perturbation is depressed during sevoflurane anesthesia. Sevoflurane-induced hypotension, which produced vagal withdrawal, did not play an important role in depressing cardio-vagal reflex function. PMID- 16368803 TI - Intrathecal and epidural anesthesia and analgesia for cardiac surgery. AB - Adequate postoperative analgesia prevents unnecessary patient discomfort. It may also decrease morbidity, postoperative hospital length of stay and, thus, cost. Achieving optimal pain relief after cardiac surgery is often difficult. Many techniques are available, and all have specific advantages and disadvantages. Intrathecal and epidural techniques clearly produce reliable analgesia in patients undergoing cardiac surgery. Additional potential benefits include stress response attenuation and thoracic cardiac sympathectomy. The quality of analgesia obtained with thoracic epidural anesthetic techniques is sufficient to allow cardiac surgery to be performed in awake patients without general endotracheal anesthesia. However, applying regional anesthetic techniques to patients undergoing cardiac surgery is not without risk. Side effects of local anesthetics (hypotension) and opioids (pruritus, nausea/vomiting, urinary retention, and respiratory depression), when used in this manner, may complicate perioperative management. Increased risk of hematoma formation in this scenario has generated much of lively debate regarding the acceptable risk-benefit ratio of applying regional anesthetic techniques to patients undergoing cardiac surgery. PMID- 16368804 TI - Spinal anesthesia in infants: is the impractical practical? PMID- 16368805 TI - The safety and efficacy of spinal anesthesia for surgery in infants: the Vermont Infant Spinal Registry. AB - Studies with modest numbers of patients have suggested that spinal anesthesia in infants is associated with a very infrequent incidence of complications, such as hypoxemia, bradycardia, and postoperative apnea. Although spinal anesthesia would seem to be a logical alternative to general anesthesia for many surgical procedures, it remains an underutilized technique. Since 1978, clinical data concerning all infants undergoing spinal anesthesia at the University of Vermont have been prospectively recorded. In all, 1554 patients have been studied. Anesthesia was performed by anesthesia trainees and attending anesthesiologists. The success rate for LP was 97.4%. An adequate level of spinal anesthesia was achieved in 95.4% of cases. The average time required to induce spinal anesthesia was 10 min. Oxygen hemoglobin desaturation to <90% was observed in 10 patients. Bradycardia (heart rate <100 bpm) occurred in 24 patients (1.6%). This study confirms the infrequent incidence of complications associated with spinal anesthesia in infants. Spinal anesthesia can be performed safely, efficiently, and with the expectation of a high degree of success. Spinal anesthesia should be strongly considered as an alternative to general anesthesia for lower abdominal and lower extremity surgery in infants. PMID- 16368806 TI - Physiological assessment and control in studies evaluating central nervous system injury: should size matter? PMID- 16368807 TI - The physiologic effects of isoflurane anesthesia in neonatal mice. AB - In neonatal rodents, isoflurane has been shown to confer neurological protection during hypoxia-ischemia and to precipitate neurodegeneration after prolonged exposure. Whether neuroprotection or neurotoxicity result from a direct effect of isoflurane on the brain or an indirect effect through hemodynamic or metabolic changes remains unknown. We recorded arterial blood pressure, heart rate, blood gases, and glucose in 10-day-old mice during 60 min of isoflurane anesthesia with spontaneous or mechanical ventilation, as well as during 60 min of hypoxia ischemia with isoflurane anesthesia or without anesthesia. During isoflurane anesthesia, hypoglycemia and metabolic acidosis occurred with spontaneous and mechanical ventilation. During hypoxia-ischemia, isoflurane was fatal with spontaneous breathing but survivable with mechanical ventilation, with arterial blood pressure and heart rate being similar to that observed in unanesthetized animals. Minimum alveolar concentration (MAC) was 2.3% in 10-day-old mice. In summary, isoflurane anesthesia precipitated hypoglycemia, which may have contributed to the neurodegeneration observed in neonatal rodents. Use of 0.8 MAC isoflurane for evaluation of neuroprotection during hypoxia-ischemia requires mechanical ventilation and glucose supplementation in this model. PMID- 16368808 TI - Predicting which child-parent pair will benefit from parental presence during induction of anesthesia: a decision-making approach. AB - Using a multiply matched, concurrent cohort analysis, with 568 subjects matched from data obtained by our laboratory over the past 7 yr, we examined whether parental presence during induction of anesthesia (PPIA) reduces children's anxiety depending on the interaction between child and parent's baseline anxiety. Children's and parents' baseline anxiety was assessed preoperatively; children's anxiety was again assessed during induction of anesthesia. We found that anxious children who received PPIA from a calm parent were significantly less anxious during induction of anesthesia as compared with anxious children who did not receive PPIA (P = 0.03). In contrast, calm children who received PPIA from an overly anxious parent were significantly more anxious as compared with calm children who were not accompanied by a parent (P = 0.002). We found no effect of PPIA on children's anxiety during induction of anesthesia when calm parents accompanied calm children into the operating room (P = 0.15) or when overly anxious parents accompanied anxious children (P = 0.49). We conclude that the presence of a calm parent does benefit an anxious child during induction of anesthesia and the presence of an overly anxious parent has no benefit. PMID- 16368809 TI - Relief of bronchial obstruction using a Fogarty catheter in a patient with bronchomalacia. AB - Tracheobronchomalacia can be latent without showing any clinical manifestations and may be incidentally found during anesthesia. In such cases, hypoxia may occur during anesthesia. We experienced obstruction of the left main bronchus caused by bronchomalacia that was incidentally found during open-heart surgery in a 4-yr old patient. We could not reopen the airway by routine techniques, such as positive pressure, and had great difficulty in weaning the patient from cardiopulmonary bypass. The use of a Fogarty catheter allowed the relief of airway obstruction and weaning from cardiopulmonary bypass. PMID- 16368810 TI - Total knee arthroplasty as an overnight-stay procedure using continuous femoral nerve blocks at home: a prospective feasibility study. AB - The average duration of hospitalization after total knee arthroplasty (TKA) in the United States is 4-5 days. In this two-phase study we investigated the feasibility of converting TKA into an overnight-stay procedure using a continuous femoral nerve block provided at home through postoperative day 4. Nine of 10 patients met discharge criteria and were discharged home the day after surgery. Pain was well controlled, opioid requirements and sleep disturbances were minimal, and patient satisfaction was high. Additional research is required to replicate these results in a controlled trial, define the appropriate subset of patients, and assess the incidence of complications associated with this practice before its mainstream use. PMID- 16368811 TI - Pharmacokinetic-pharmacodynamic modeling the hypnotic effect of sevoflurane using the spectral entropy of the electroencephalogram. AB - Spectral entropy is a new electroencephalogram (EEG)-derived parameter that may be used to model the pharmacokinetic-pharmacodynamic (PKPD) effects of general anesthetics. In the present study we sought to derive a PKPD model of the relationship between sevoflurane concentration and spectral entropy of the EEG. We collected spectral entropy data during increasing and decreasing sevoflurane anesthesia from 20 patients. The first cycle consisted of induction and lightening phases with no supplemental medications. An effect-site compartment and inhibitory E(max) model described the relation between sevoflurane concentration and spectral entropy. PKPD parameters were derived from the full cycle and separately from the increasing and decreasing stages. The second anesthetic cycle consisted of a redeepening phase only and included airway manipulation and routinely administered adjunctives. PKPD data obtained from the first cycle were used to predict second cycle entropy changes. There was a consistent relationship between effect-site sevoflurane concentration and spectral entropy (median absolute weighted residual = 11.6%). For complete first cycle response entropy (mean +/- sd): T1/2 K(eo) = 2.4 +/- 1.5 min, gamma = 5.9 +/- 2.3, EC50 = 1.7 +/- 0.3. We found significant differences between gamma values when the sevoflurane concentration was increasing (61.1 +/- 55.2) compared with the decreasing part of the cycle (5.7 +/- 2.8). Above an effect-site concentration of 3%, spectral entropy of the EEG is unresponsive to further increases in sevoflurane concentration. The effect-compartment inhibitory E(max) model accurately describes the relation between sevoflurane concentration and spectral entropy of the EEG. Spectral entropy decreases with increasing sevoflurane concentrations up to 3%. The steepness of the dose-response curve varies between phases of increasing and decreasing anesthetic concentrations. PMID- 16368812 TI - The effects of sevoflurane and propofol on QT interval and heterologously expressed human ether-a-go-go related gene currents in Xenopus oocytes. AB - Sevoflurane can induce prolongation of the cardiac QT interval by inhibiting the repolarization phase of the action potential. This may occur as a result of inhibition of the human ether-a-go-go related gene (HERG) channel. To clarify the mechanisms of anesthetics on HERG channels, we monitored the electrocardiogram and measured QT intervals in the guinea pig in the presence of sevoflurane and propofol. Sevoflurane (1%-4%) prolonged QTc dose-dependently (7.5%-21.2%), but propofol did not affect it. Furthermore, HERG channels were expressed in Xenopus oocytes and outward HERG currents were obtained on step depolarization from a holding potential of -70 mV. Repolarization to -70 mV from positive test potentials resulted in large outward tail currents. Sevoflurane (1%-4%), in a dose-dependent manner, inhibited the HERG outward tail currents (9.7%-26.6%), whereas steady-state currents were inhibited only at large concentrations. The time constant of the converging current was decreased in the presence of sevoflurane, but the inactivation and activation curves were not shifted. Propofol did not affect these currents within the clinically relevant concentration. In conclusion, compared with steady-state currents, sevoflurane was more potent in inhibiting the outward tail currents, suggesting that sevoflurane may modulate the HERG channel kinetics in its inactivated state. PMID- 16368813 TI - A novel molecule with peripheral opioid properties: the effects on hypercarbic and hypoxic ventilation at steady-state compared with morphine and placebo. AB - Frakefamide (FF), is a new peripherally acting mu-opioid receptor agonist. The aim of this double-blind, randomized, double-dummy, four-way, crossover study was to investigate FF effects on hypercarbic and hypoxic ventilation at steady-state after a 6-h infusion. We compared the effect with 2 clinical doses of morphine (M small and M-large) and placebo in 12 healthy men. The subjects received 1.22 mg/kg of FF, 0.44 mg/kg of M-large, and 0.11 mg/kg of M-small. Sodium chloride 9 mg/mL was used as placebo. Ventilation was studied by pneumotachography and in line capnography. There were no ventilatory effects caused by FF or placebo. As expected, large doses of morphine influenced both hypercarbic and hypoxic ventilatory responses. We conclude that there were no signs of central respiratory depression caused by FF after 6 h of constant infusion, which supports a peripheral action of the compound. However, morphine caused a dose dependent central depression during the hypercarbic ventilatory response and a mild depression of hypoxic ventilatory response. PMID- 16368814 TI - The effect of nefopam on morphine overconsumption induced by large-dose remifentanil during propofol anesthesia for major abdominal surgery. AB - Opioids may activate pain facilitatory systems opposing analgesia. We investigated whether large-dose remifentanil given during IV anesthesia caused postoperative morphine overconsumption and whether nefopam (a centrally acting analgesic) could reduce this. Sixty patients scheduled for abdominal surgery were included in this prospective, randomized study. The first 30 patients received either small-dose (Group S: 3 ng/mL) or large-dose (Group L: 8 ng/mL) remifentanil administrated by a target-controlled infusion during propofol anesthesia. Before skin closure, patients received morphine 0.15 mg/kg. Another 30 patients also received nefopam 20 mg intraoperatively. Postoperative pain was controlled by titration of morphine, followed by patient-controlled morphine analgesia (PCA). Morphine was requested earlier in Group L than in Group S (10 [1 63] min versus 37 [5-90] min, median [range]; P < 0.002). The dose of morphine by titration was larger in Group L than in Group S (0.28 [0.04-0.38] mg/kg versus 0.16 [0.03-0.41] mg/kg; P < 0.05). PCA morphine consumption and pain scores were similar. There were no differences between the nefopam groups in the time to first morphine request or in the dose of morphine by titration. Postoperative morphine overconsumption occurred after large-dose remifentanil and propofol anesthesia during the early postoperative period. Pretreatment with nefopam could be useful to prevent pain sensitization induced by opioids. PMID- 16368815 TI - An analysis of remifentanil in the pulmonary vascular bed of the cat. AB - In this investigation we sought to identify the role of remifentanil in the feline pulmonary vascular bed. Using adult mongrel cats in separate experiments, the effects of glibenclamide (adenosine triphosphate-sensitive K+ channel blocker), diphenhydramine (histamine H(1)-receptor antagonist), L-N5-(1 Iminoethyl) ornithine hydrochloride (nitric oxide synthase inhibitor), and naloxone (opioid receptor antagonist) were investigated in pulmonary arterial responses to remifentanil (opioid agonist), pinacidil (adenosine triphosphate sensitive K+ channel activator), and bradykinin (nitric oxide synthase inducer). Under increased tone conditions in the isolated left lower lobe vascular bed of the cat, remifentanil induced a dose-dependent vasodepressor response that was not significantly altered after administration of glibenclamide and L-N5-(1 Iminoethyl) ornithine hydrochloride. Responses to remifentanil were significantly attenuated after administration of diphenhydramine and naloxone. The results suggest that remifentanil has potent vasodepressor activity in the feline pulmonary vascular bed and that these responses are mediated by histamine and opioid receptor sensitive pathways. PMID- 16368816 TI - Actions of norepinephrine and isoflurane on inhibitory synaptic transmission in adult rat spinal cord substantia gelatinosa neurons. AB - Volatile inhaled anesthetics and nitrous oxide (N2O) are often used together in clinical practice to produce analgesia. Because the analgesic effect of N2O is, at least in part, mediated by norepinephrine (NE) release in the spinal cord, we examined the interaction between isoflurane (ISO) and NE in the adult rat spinal cord with respect to central nociceptive information processing. The effects of clinically relevant concentrations of ISO (1 MAC) and NE (20 microM) on spontaneous inhibitory transmission in substantia gelatinosa (SG) neurons were examined using the blind whole-cell patch-clamp method. ISO prolonged the decay time and increased the total charge transfer of spontaneous inhibitory postsynaptic currents. NE increased the frequency and mean amplitude of inhibitory postsynaptic currents and the charge transfer as well. Coapplication of both drugs led to an additive increase of the charge transfer and frequent temporal summation of inhibitory postsynaptic currents. We conclude that both ISO and NE enhance the inhibitory synaptic transmission in the rat SG neurons and their interaction is additive, suggesting that ISO may add to the analgesic action of N2O at the spinal cord dorsal horn level. PMID- 16368817 TI - The efficacy and safety of intravenous emulsified isoflurane in rats. AB - Although direct IV injection of liquid volatile anesthetics is usually lethal, anesthesia using emulsified halothane and isoflurane without adverse effects has been safely induced in animals. We identified the safe concentration of emulsified volatile anesthetic preparations and determined the dose-response relationship of IV emulsified isoflurane and propofol in rats. Liquid/gas partition coefficients of desflurane, sevoflurane, isoflurane, enflurane, and halothane in 20% and 30% Intralipid were measured and used to calculate their saturated concentrations. Unsaturated emulsified isoflurane was prepared by adding liquid isoflurane to 30% Intralipid. The loss of forepaw righting reflex was taken as induction of anesthesia, and disappearance of electrocardiogram was taken as death. The median effective induction dose (ED50) and median lethal dose (LD50) of emulsified isoflurane were 0.072 and 0.216 mL/kg liquid isoflurane, respectively. The ED50 and LD50 of propofol were 5.89 mg/kg and 18.19 mg/kg, respectively. Time to return of forepaw righting reflex after injection of emulsified isoflurane (38 +/- 18 s) was significantly shorter than with propofol (101 +/- 62 s; P < 0.05). Anesthesia was successfully induced in rats by IV emulsified isoflurane with a comparable safety index and certain safety factor as propofol. Recovery of anesthesia after IV emulsified isoflurane was faster than with propofol. PMID- 16368818 TI - Cyclooxygenase-1 inhibition shortens the duration of diazepam-induced loss of righting reflex in mice. AB - Cyclooxygenase-1 (COX-1) inhibition by a selective inhibitor valeryl salicylate, or nonselective inhibitors at 10 mg/kg, including aspirin, ibuprofen, indomethacin, and picroxicam, attenuated by 29%-46% the duration of loss of righting reflex induced by diazepam (20 mg/kg) in mice. On the other hand, arachidonic acid (20 mg/kg) increased the duration of diazepam-induced loss of righting reflex by 48%. This effect of arachidonic acid was abolished by aspirin. However, aspirin at 10 mg/kg also did not alter the effects of diazepam (5 mg/kg) on spontaneous activity and rotarod performance. These findings strongly suggest that one or more COX products, most likely prostaglandins, play a significant role in modulating the hypnotic effect of diazepam. Elucidating the mechanism involved may further our understanding of the pharmacology of benzodiazepines. PMID- 16368819 TI - The local anesthetic butamben inhibits and accelerates low-voltage activated T type currents in small sensory neurons. AB - Butamben (BAB) is a local anesthetic that can be used in epidural suspensions for long-term selective suppression of dorsal root pain signal transmission and in ointments for the treatment of skin pain. Previously, high-voltage activated N type calcium channel inhibition has been implicated in the analgesic effect of BAB. In the present study we show that low-voltage activated or T-type calcium channels may also contribute to this effect. Typical transient T-type barium currents, selectively evoked by low-voltage (-40 mV) clamp stimulation of small (approximately 20 microm diameter) dorsal root ganglion neurons from newborn mice, were inhibited by BAB with an IC50 value of approximately 200 microM. Furthermore, 200 microM BAB accelerated T-type current activation, deactivation, and inactivation kinetics, comparable to earlier observations for N-type calcium channels. Finally, 200 microM BAB had no effect on the midpoint potential and slope factor of the activation curve, although it caused a approximately 3 mV hyperpolarizing shift of the inactivation curve, without affecting the slope factor. We conclude that BAB inhibits T-type calcium channels with a mechanism associated with channel kinetics acceleration. PMID- 16368820 TI - The analgesic effect of tramadol after intravenous injection in healthy volunteers in relation to CYP2D6. AB - Tramadol analgesia results from a monoaminergic effect by tramadol itself and an opioid effect of its metabolite (+)-M1 formed by O-demethylation of tramadol by CYP2D6. In this study we sought to determine the impact of (+)-M1 on the analgesic effect of tramadol evaluated by experimental pain models. The effect of an IV injection of 100 mg tramadol on experimental pain was studied 15-90 min after dosing in volunteers, 10 extensive metabolizers with CYP2D6 and 10 poor metabolizers without CYP2D6 in 2 placebo-controlled trials. The pain tests included detection and tolerance threshold to single electrical sural nerve stimulation, pain summation threshold to repetitive electrical sural nerve stimulation (temporal summation), and the cold pressor test. In extensive metabolizers, tramadol reduced discomfort experienced during the cold pressor test (P = 0.002). In poor metabolizers, the pain tolerance thresholds to sural nerve stimulation were increased (P = 0.04). (+)-M1 could be detected in the serum samples from all extensive metabolizers except one, but (+)-M1 was below the limit of determination in all poor metabolizers. The opioid effect of (+)-M1 appears to contribute to the analgesic effect of tramadol, but the monoaminergic effect of tramadol itself seems to create an analgesic effect. PMID- 16368821 TI - The dose of succinylcholine required for excellent endotracheal intubating conditions. AB - In this prospective, randomized, double-blind, placebo-controlled study, we attempted to define the dose of succinylcholine that provides excellent intubation conditions in patients within 60 s during simulated rapid-sequence induction of anesthesia. Anesthesia was induced in 180 patients with 2 microg/kg fentanyl and 2 mg/kg propofol. After loss of consciousness, patients were randomly allocated to receive 0.3, 0.5, 1.0, 1.5, or 2.0 mg/kg succinylcholine or saline solution (control group). Tracheal intubation was performed 60 s later. A blinded investigator performed all laryngoscopies and graded intubating conditions. Intubating conditions were excellent in 0.0%, 43.3%, 60.0%, 63.3%, 80.0%, and 86.7% of patients after 0.0, 0.3, 0.5, 1.0, 1.5, and 2.0 mg/kg succinylcholine, respectively. The incidence of excellent intubating conditions was significantly more frequent (P < 0.001) in patients receiving succinylcholine than in the controls and in patients who received 2.0 mg/kg succinylcholine (P < 0.05) than in those who received 0.3 mg/kg succinylcholine. The calculated doses of succinylcholine (and their 95% confidence intervals) that are required to achieve excellent intubating conditions in 50% and 80% of patients at 60 s are 0.39 (0.29-0.51) mg/kg and 1.6 (1.2-2.0) mg/kg, respectively. It appears that there are no advantages to using doses of succinylcholine larger than 1.5 mg/kg. PMID- 16368822 TI - Lidocaine does not prevent bispectral index increases in response to endotracheal intubation. AB - We investigated the effect of IV lidocaine on the hemodynamic and bispectral index responses to induction of general anesthesia and endotracheal intubation. Forty patients (ASA I) were randomly allocated into 2 groups of 20 to receive normal saline or lidocaine 1.5 mg/kg IV 30 s after induction. Ninety seconds later, endotracheal intubation was performed. Systolic blood pressure, heart rate, and bispectral index were measured at baseline, 1 min after induction, at preintubation, and every minute until 5 min after endotracheal intubation. Bispectral index at 1 min after induction and preintubation in the lidocaine group were significantly lower compared with the control group (P < 0.05). Systolic blood pressure increased significantly at 1 and 2 min after intubation in the control group compared with the baseline value (P < 0.05) but did not increase significantly in the lidocaine group. Heart rate increased at 1 to 3 min in both groups (P < 0.05), but there was no significant difference between the two groups. One patient in the control group had recall of the procedure. We conclude that the administration of IV lidocaine (1.5 mg/kg) does not suppress the hypnotic response to endotracheal intubation. PMID- 16368823 TI - A comparison of state and response entropy versus bispectral index values during the perioperative period. AB - Cerebral monitoring indices are associated with a large degree of inter-patient variability and electrical signal interference during surgery. We designed this clinical study to test the hypothesis that use of the spectral entropy (Entropy) module is associated with less frequent intraoperative interference with the displayed indices than the bispectral index (BIS) monitor when used during general anesthesia with propofol and desflurane. Thirty consenting patients scheduled for major laparoscopic surgery procedures were enrolled in this prospective study. The elapsed time to obtain a baseline index value was recorded, as well as the simultaneous state entropy (SE), response entropy (RE), and BIS values at specific time intervals during the induction, maintenance, and emergence periods in patients administered a standardized general anesthetic technique. During the maintenance period, the changes in these indices were evaluated after a bolus dose of propofol (20 mg IV) and a 2% increase or decrease in the inspired concentration of desflurane. As expected, the baseline SE values were less than the RE and BIS values (88 +/- 2 versus 96 +/- 3 and 96 +/- 4, respectively). However, the SE and RE values correlated with the BIS value during the induction (r = 0.77 and 0.78, respectively) and emergence (r = 0.86 and 0.91, respectively) periods. The area under the receiver operating characteristic curve for detection of consciousness also indicated a similar performance of the SE (0.93 +/- 0.04) relative to the RE (0.98 +/- 0.04) and BIS (0.97 +/- 0.04). During the maintenance period, the responses to changes in propofol and desflurane concentrations were consistent with all three indices. Finally, the entropy indices were less interfered with by the electrocautery unit during the operation (12% versus 62% for the BIS monitor). Because the average selling prices of the Entropy and BIS disposable electrode strips (14.25 dollars versus 14.95 dollars USD, respectively) are comparable, we conclude that the Entropy module is a cost-equivalent alternative to the BIS monitor. PMID- 16368824 TI - An audible indication of exhalation increases delivered tidal volume during bag valve mask ventilation of a patient simulator. AB - Self-inflating manual resuscitators (SIMRs) can mislead caregivers because the bag, unlike a Mapleson-type device, reinflates even without patient exhalation. We added a whistle as an audible indicator to the exhalation port of a SIMR. In randomized order, each participant provided two sets of breaths via mask ventilation with a SIMR, one with and one without audible feedback, to a Human Patient Simulator modified to log lung volume changes. The last three breaths in each set were used to compare average tidal volume (Vt) under both conditions. Eighty-seven advanced cardiac life support trainees (54 males, 33 females) with clinical experience averaging 6.4 +/- 9.4 yr were recruited. Average Vt delivered with the standard SIMR was 486 +/- 166 mL and 624 +/- 96 mL with the modified SIMR. Average Vt delivered by a modified SIMR was significantly larger by 40% when it followed standard SIMR use and 19% when using the modified SIMR first. Use of a SIMR with an audible indicator of exhalation significantly (P < 0.001) increased mask ventilation of a patient simulator, suggesting that mask ventilation of a patient with a SIMR may also be increased by objective, real time feedback of exhaled Vt. PMID- 16368825 TI - Fires in the operating room and intensive care unit: awareness is the key to prevention. AB - Recent recommendations from the Centers for Disease Control (CDC) to use alcohol based substances for hand hygiene and skin antisepsis could introduce new fire hazards in the operating room (OR). This potential for an increase in the number of fires in the hospital setting with wide spread use of alcohol-based agents warrants heightened awareness of the risks and implementation of safety measures when using these agents. Here, we report a patient who, during a tracheostomy, sustained severe burns resulting from a fire in the OR. In this case, the use of an alcohol-based antiseptic was the major contributing factor to the surgical fire. PMID- 16368826 TI - Gabapentin: an alternative to the cyclooxygenase-2 inhibitors for perioperative pain management. AB - The cyclooxygenase-2 inhibitor, rofecoxib, was a popular analgesic adjuvant for improving perioperative pain management. We designed this placebo-controlled study to test the hypothesis that gabapentin could produce similar reductions in postoperative pain and opioid analgesic usage, thereby improving the recovery process. One hundred patients undergoing abdominal hysterectomy procedures were randomly assigned to one of four treatment groups: 1) control group received placebo capsules and pills before and for 2 days after surgery, 2) rofecoxib group received 50 mg/d PO and placebo capsules before and after surgery and, 3) gabapentin group received 1.2 g/d PO and placebo pills before and after surgery, and 4) combination group received rofecoxib 50 mg/d and gabapentin 1.2 g/d PO before and after surgery. The anesthetic technique was standardized and the postoperative assessments included verbal rating scales for pain and sedation, IV morphine usage, quality of recovery assessment, recovery of bowel function, resumption of normal activities, and patient satisfaction with their pain management. Postoperative pain scores were significantly reduced in all three analgesic treatment groups (versus control group). Compared with the control group, patient-controlled analgesia morphine usage was also significantly reduced in the 3 analgesic treatment groups at 1, 8, 24, and 30 h after surgery. Total PCA morphine usage was decreased by 43%, 24%, and 50% in groups 2, 3, and 4, respectively, compared with group 1. Oral analgesic consumption was also smaller in groups 2 and 4 when compared with the control group. The opioid-sparing effects of rofecoxib and gabapentin lead to a faster recovery of bowel function. Discharge eligibility scores in groups 2 and 4 were improved at 24 h when compared with group 1, and patient satisfaction with postoperative pain management was significantly higher at 24 h in all 3 analgesic treatment groups. At the 72 h follow-up, all of the patients in group 4 were completely satisfied with their pain management compared with only 32%, 64%, and 72% in groups 1, 2, and 3, respectively. Gabapentin (1.2 g/d PO) appears to be an acceptable alternative to rofecoxib (50 mg/d PO) for short-term use as an adjuvant to opioid analgesics in patients undergoing lower abdominal surgery. PMID- 16368827 TI - The antiallodynic action target of intrathecal gabapentin: Ca2+ channels, KATP channels or N-methyl-d-aspartic acid receptors? AB - Gabapentin is a novel analgesic whose mechanism of action is not known. We investigated in a postoperative pain model whether adenosine triphosphate (ATP) sensitive K+ (K(ATP)) channels, N-methyl-d-aspartic acid (NMDA) receptors, and Ca2+ channels are involved in the antiallodynic effect of intrathecal gabapentin. Mechanical allodynia was induced by a paw incision in isoflurane-anesthetized rats. Withdrawal thresholds to von Frey filament stimulation near the incision site were measured before and after incision and after intrathecal drug administration. The antiallodynic effect of gabapentin (100 mug) was not affected by intrathecal pretreatment with antagonists of K(ATP) channels, NMDA receptors or gamma-aminobutyric acid (GABA)(A) receptors. K(ATP) channel openers and GABA(A) receptor agonist, per se, had little effect on the postincision allodynic response. The Ca2+ channel blocker of N-type (omega-conotoxin GVIA, 0.1-3 microg), but not of P/Q-type (omega-agatoxin IVA), L-type (verapamil, diltiazem or nimodipine), or T-type (mibefradil), attenuated the incision-induced allodynia, as did gabapentin. Both the antiallodynic effects of gabapentin and omega-conotoxin GVIA were attenuated by Bay K 8644, an L-type Ca2+ channel activator. These results provide correlative evidence to support the contention that N-type Ca2+ channels, but not K(ATP) channels or NMDA or GABA(A) receptors, might be involved in the antiallodynic effect of intrathecal gabapentin. PMID- 16368828 TI - An iontophoretic fentanyl patient-activated analgesic delivery system for postoperative pain: a double-blind, placebo-controlled trial. AB - An iontophoretic fentanyl HCl patient-activated transdermal system (fentanyl HCl PATS) is under development for the treatment of acute postoperative pain. The fentanyl HCl PATS is a needle-free, credit card-sized, preprogrammed system that is applied to the patient's upper outer arm or chest. The fentanyl HCl PATS was demonstrated to be superior to placebo in a previous trial; however, the randomization scheme used and the lack of control of entry pain level may have contributed to the lack of robust findings. We compared the fentanyl HCl PATS with placebo for acute postoperative pain management in a larger trial that addressed the limitations of the previous study. Adult patients admitted to the postanesthesia care unit after major surgery were titrated to comfort with opioids and randomized 1:1 to receive the fentanyl HCl PATS 40 microg or placebo for 24 hours. Supplemental IV fentanyl was available to patients upon request in both treatment groups for the first 3 hours after enrollment. The primary efficacy end-point was the percentage of patients who discontinued participation in the study because of inadequate analgesia. Pain intensity scores, patient global assessments (PGA), and investigator global assessments (IGA) were collected. Four-hundred-eighty-four patients (PATS, n = 244; placebo, n = 240) were enrolled. Fewer patients receiving the fentanyl HCl PATS discontinued because of inadequate analgesia compared with placebo (28.7% versus 60.0%; P < 0.0001). Mean last pain intensity scores were 3.5 and 5.4 for the fentanyl HCl PATS and placebo groups, respectively. Patients (73.4%, PGA) and investigators (72.1%, IGA) considered the fentanyl HCl PATS a good or excellent method of pain control. Treatment-related adverse events were similar between groups. This study demonstrated the superiority of the iontophoretic fentanyl HCl PATS over placebo for acute postoperative pain management. PMID- 16368829 TI - Nonsteroidal antiinflammatory drugs suppress pain-related behaviors, but not referred hyperalgesia of visceral pain in mice. AB - Visceral pain is characterized by spontaneous pain and referred hyperalgesia. After inducing visceral pain in mice using intracolonic mustard oil administration, we examined the effects of various nonsteroidal antiinflammatory drugs (NSAIDs) on pain-related behavior and on Evans blue dye extravasation. Animals were given one of the following: saline, ethanol, dimethylsulfoxide (DMSO), morphine, ketoprofen, ketorolac, or DFU (a cyclooxygenase-2 inhibitor). After drug treatment, mice underwent intracolonic administration of 50 microL 1.5% mustard oil. Spontaneous pain-related responses were assessed for the next 20 min. The frequency of withdrawal responses to the application of von Frey hairs to the abdomen, foot, and tail was determined. After completion of the behavioral tests, Evans blue was injected into the animals via the tail vein. Two hours later, the colon was removed postmortem and Evans blue content was measured. Spontaneous pain behaviors were significantly less in animals administered 3 and 10 mg/kg morphine, 50 mg/kg ketorolac, 100 mg/kg ketoprofen, and 20 mg/kg DFU (P < 0.05). Response frequencies to the application of von Frey hairs were lower in mice administered 3 and 10 mg/kg morphine (P < 0.05) but were not affected by ketorolac, ketoprofen, or DFU treatment. Colonic Evans blue content was smaller in mice given 100 mg/kg ketoprofen and 20 mg/kg DFU (P < 0.05). We concluded that NSAIDs reduced pain behavior and inflammation but had little effect on referred hyperalgesia. PMID- 16368830 TI - The efficacy of thoracic epidural neostigmine infusion after thoracotomy. AB - Few anesthesia studies have explored perioperative continuous epidural infusion of neostigmine. We examined such a regimen in thoracotomy patients. Ninety patients were randomized to one of three groups in this double-blind trial. Before anesthesia induction, an epidural catheter was inserted in all patients at T5-8 levels under local anesthesia. Pre-neo patients received bolus 500-microg epidural neostigmine before anesthesia induction followed by infusion of 125 microg/h until the end of surgery. Post-neo patients received epidural saline during the same time periods plus bolus 500-microg epidural neostigmine at end of surgery. Patients in the control group received saline placebo during all three periods. Patients in the neostigmine groups postoperatively received patient controlled epidural analgesia with morphine 0.02 mg/mL, bupivacaine 0.08 mg/mL, and neostigmine 7 microg/mL. Control patient-controlled epidural analgesia excluded neostigmine. Data were recorded for 6 postoperative days. Daily patient controlled epidural analgesia consumption (mL) for Pre-neo patients was significantly less than that of post-neo and control group patients for postoperative days 1-6 (at least 10% and 16% less, respectively; P < 0.05). There was a modest decrease in pain intensity on postoperative days 3-6 for pre-neo patients versus other groups (P < 0.05). These results suggest that continuous thoracic epidural neostigmine started before anesthesia provided preemptive, preventive analgesia and an analgesic-sparing effect that improved postoperative analgesia for these patients without increasing the incidence of adverse effects. PMID- 16368831 TI - Postgraduate medical education: how do we know it's working? PMID- 16368832 TI - Accreditation Council for Graduate Medical Education competencies and the American Board of Anesthesiology Clinical Competence Committee: a comparison. AB - We compared the Accreditation Council for Graduate Medical Education (ACGME) Outcome Project to the long-standing requirement of the American Board of Anesthesiology for a Clinical Competence Committee Report. There are many similarities between these two systems of resident evaluation. However, the ACGME Outcome Project requires the use of more numerous and diverse metrics when compared with the traditional global evaluation alone. In addition, the Clinical Competence Committee Report is primarily a summative evaluation for the purpose of assigning credit for training. The ACGME Outcome Project may be used as a component of a summative evaluation, but the primary emphasis is on formative assessment. PMID- 16368833 TI - Outcomes of cardiopulmonary resuscitation and predictors of survival in patients undergoing coronary angiography including percutaneous coronary interventions. AB - We studied the outcome of cardiopulmonary resuscitation (CPR) in patients undergoing coronary angiography (CA) and/or percutaneous coronary interventions (PCI). Of 51,985 CA and PCI patients treated between January 1, 1990, and December 31, 2000, 114 required CPR. Records were reviewed for relationships between patient characteristics and various procedures and short-term survival. Long-term survival was compared with that of a matched cohort of patients who did not have an arrest during catheterization and a matched cohort from the general Minnesota population. Over the 11-year period, the overall incidence of CPR was 21.9 per 10,000 procedures. This rate decreased from 33.9 per 10,000 before 1995 to 13.1 per 10,000 after 1995. Overall survival to hospital discharge after CPR was 56.1%. Survival to discharge was less frequent with a history of congestive heart failure, previous coronary artery bypass graft surgery, hemodynamic instability during the procedure, and with prolonged or emergent catheterizations. Pulseless electrical activity (versus asystole or ventricular fibrillation) indicated very poor short-term survival. Interestingly, short-term survival was not related to the extent of coronary artery disease. Long-term survival of patients who survived cardiac arrest was comparable to that of those who did not have arrest during catheterization. In conclusion, the incidence of periprocedural CPR during diagnostic or interventional coronary procedures decreased after 1995. Patients who received CPR in the cardiac catheterization lab have a remarkably frequent survival to hospital discharge rate. Long-term survival of these patients is only minimally reduced. PMID- 16368834 TI - Pretreatment with simvastatin reduces lung injury related to intestinal ischemia reperfusion in rats. AB - In this rat model study we evaluated whether pretreatment with simvastatin affects the severity of acute lung injury caused by intestinal ischemia reperfusion (I/R). Twenty-four animals were randomly allocated to three equal groups (sham, control, simvastatin). The simvastatin group was pretreated with simvastatin 10 mg x kg(-1) x day(-1) for 3 days, whereas the other groups received placebo. The simvastatin and control groups underwent 60 min of superior mesenteric artery occlusion and 90 min of reperfusion. Compared with the simvastatin group, the control group exhibited significantly more severe intestinal I/R-induced acute lung injury, as indicated by lower Pao2 and oxygen saturation (P = 0.01 and P = 0.005, respectively) and higher mean values for neutrophil infiltration of the lungs (P = 0.003), total lung histopathologic injury score (P = 0.003), lung wet-to-dry weight ratio (P = 0.009), and lung tissue malondialdehyde levels (P = 0.016). The control and simvastatin groups had similar serum levels and similar bronchoalveolar lavage fluid levels of cytokines (interleukin-1, interleukin-6, and tumor necrosis factor-alpha) and P-selectin at all measurements, except for a significantly higher level of bronchoalveolar lavage fluid P-selectin in the control group (P = 0.006). Pretreatment with simvastatin reduces the severity of acute lung injury induced by intestinal I/R in rats. PMID- 16368835 TI - Unmasking of Brugada syndrome by an antiarrhythmic drug in a patient with septic shock. AB - Asymptomatic Brugada syndrome patients often display concealed Brugada-type electrocardiogram patterns that result in under-diagnosis of this syndrome. These patients include individuals of both genders and a wide range of ages. They are as likely as non-Brugada patients to have normal longevity or to suffer from a critical illness. Here we report a case of septic shock in which Brugada-type electrocardiogram patterns were induced by pilsicainide administration for the treatment of atrial fibrillation. This case report suggests that some drugs used in the treatment of septic shock can unmask the Brugada-type electrocardiogram pattern and induce lethal ventricular tachyarrhythmia. PMID- 16368836 TI - Patient satisfaction with awake craniotomy for tumor surgery: a comparison of remifentanil and fentanyl in conjunction with propofol. AB - In this study we compared the effectiveness of the use of remifentanil to fentanyl in conjunction with propofol in providing conscious sedation for awake craniotomy for tumor surgery and to assess patient satisfaction with both techniques. The ability to maintain appropriate levels of sedation, adequate analgesia, and hemodynamic stability was assessed in 50 patients randomized to receive either fentanyl or remifentanil. All complications were documented. Patients were interviewed at 1 h, 4 h, and 24 h after surgery to note their recall of procedure and pain and their overall satisfaction. There were no differences in sedation and pain scores or in hemodynamic and respiratory variables between the two groups. The incidence of intraoperative complications was not different (fentanyl, 14; remifentanil, 16). Respiratory complications occurred in 9 (18%) patients (fentanyl 6, remifentanil 3). The recall and satisfaction scores were not different; 93% of all patients were completely satisfied at all interview times. The use of remifentanil infusion in conjunction with propofol is a good alternative to fentanyl and propofol for conscious sedation for the awake craniotomy and these techniques are both well accepted by the patient. PMID- 16368837 TI - The sitting versus right lateral position during combined spinal-epidural anesthesia for cesarean delivery: block characteristics and severity of hypotension. AB - In the present study we evaluated whether the sitting position during initiation of small-dose combined spinal-epidural anesthesia (CSE) would induce less hypotension as compared with the lateral position. Sixty women undergoing elective cesarean delivery were randomly assigned to receive a spinal injection consisting of 6.6 mg hyperbaric bupivacaine with sufentanil 3.3 microg in either the lateral or the sitting position. After securing the epidural catheter, patients were turned to a 15 degrees left lateral supine position. Ephedrine 5 mg IV was administered prophylactically and subsequently in case of nausea/vomiting and/or hypotension, defined as a systolic blood pressure less than 95 mm Hg or a 25% decrease from baseline values. Although the incidence of ephedrine supplementation was not different, females in the sitting group required less ephedrine (P = 0.012) and there were fewer problems with identifying the epidural space (P = 0.01). However, more patients in this group required epidural supplementation (35% versus 3%; P = 0.007). In the lateral group, blocks extended more cephalad than with the sitting position (P = 0.014). Apgar scores did not differ, but umbilical artery pH values were significantly higher in patients of the sitting group (7.31 +/- 0.04 versus 7.26 +/- 0.03; P = 0.02). We conclude that performing a CSE technique for cesarean delivery in the sitting position was technically easier and induced less severe hypotension. PMID- 16368838 TI - Does continuous peripheral nerve block provide superior pain control to opioids? A meta-analysis. AB - Although most randomized clinical trials conclude that the addition of continuous peripheral nerve blockade (CPNB) decreases postoperative pain and opioid-related side effects when compared with opioids, studies have included relatively small numbers of patients and the majority failed to show statistical significance during all time periods for reduced pain or side effects. We identified studies primarily by searching Ovid Medline (1966-May 21, 2004) for terms related to postoperative analgesia with CPNB and opioids. Each article from the final search was reviewed and data were extracted from tables, text, or extrapolated from figures as needed. Nineteen articles, enrolling 603 patients, met all inclusion criteria. Inclusion criteria were a clearly defined anesthetic technique (combined general/regional anesthesia, general anesthesia alone, peripheral nerve block), randomized trial, adult patient population (> or =18 yr old), CPNB (or analgesia) used postoperatively (intrapleural catheters were deemed not to be classified as a peripheral nerve catheter), and opioids administered for postoperative analgesia in groups not receiving peripheral nerve block. Perineural analgesia provided better postoperative analgesia compared with opioids (P < 0.001). This effect was seen for all time periods measured for both mean visual analog scale and maximum visual analog scale at 24 h (P < 0.001), 48 h (P < 0.001), and 72 h (mean visual analog scale only) (P < 0.001) postoperatively. Perineural catheters provided superior analgesia to opioids for all catheter locations and time periods (P < 0.05). Nausea/vomiting, sedation, and pruritus all occurred more commonly with opioid analgesia (P < 0.001). A reduction in opioid use was noted with perineural analgesia (P < 0.001). CPNB analgesia, regardless of catheter location, provided superior postoperative analgesia and fewer opioid-related side effects when compared with opioid analgesia. PMID- 16368839 TI - Stimulating popliteal catheters for postoperative analgesia after hallux valgus repair. AB - Stimulating catheters have been recently introduced in clinical practice. We assessed the efficacy of stimulating and nonstimulating catheter placement for pain control and local anesthetic requirements after hallux valgus repair with continuous sciatic popliteal nerve block in this comparative, randomized, blinded to-observer study of 48 patients. A stimulating catheter was placed in groups S 125 and S-0625. The same catheter was inserted without stimulation in group NS 125. An infusion of 0.125% levobupivacaine was given in groups S-125 and NS-125, whereas 0.0625% levobupivacaine was used in group S-0625. All patients received an infusion of the test drug at a basal rate of 3 mL/h, with the possibility of an additional bolus of 3 mL every hour. Verbal analog scale (VAS) scores for pain were assessed between 6-8 h and between 19-23 h postoperatively. Multiple attempts were required for catheter insertion in all patients in groups S-125 and S-0625. Lower median (range) VAS scores for pain (0-100 points) were found in group S-125 at 6-8 h postoperatively when compared with groups S-0625 and NS-125: 5 (0-17.5) versus 60 (15-80) and 70 (25-80), respectively (P < 0.05); and lower VAS scores for pain were also found in group S-125 at 19-23 h when compared with group NS-125: 0 (0-0) and 7.5 (0-10), respectively (P < 0.05). Fewer patients required IV opioid analgesia in group S-125 than in groups S-0625 and NS-125: 0, 5, and 7 patients, respectively (P < 0.05). We conclude that efficacy in pain control was increased with stimulating catheter placement. PMID- 16368840 TI - Dexamethasone added to lidocaine prolongs axillary brachial plexus blockade. AB - Different additives have been used to prolong regional blockade. We designed a prospective, randomized, double-blind study to evaluate the effect of dexamethasone added to lidocaine on the onset and duration of axillary brachial plexus block. Sixty patients scheduled for elective hand and forearm surgery under axillary brachial plexus block were randomly allocated to receive either 34 mL lidocaine 1.5% with 2 mL of isotonic saline chloride (control group, n = 30) or 34 mL lidocaine 1.5% with 2 mL of dexamethasone (8 mg) (dexamethasone group, n = 30). Neither epinephrine nor bicarbonate was added to the treatment mixture. We used a nerve stimulator and multiple stimulations technique in all of the patients. After performance of the block, sensory and motor blockade of radial, median, musculocutaneous, and ulnar nerves were recorded at 5, 15, and 30 min. The onset time of the sensory and motor blockade was defined as the time between last injection and the total abolition of the pinprick response and complete paralysis. The duration of sensory and motor blocks were considered as the time interval between the administration of the local anesthetic and the first postoperative pain and complete recovery of motor functions. Sixteen patients were excluded because of unsuccessful blockade. The duration of surgery and the onset times of sensory and motor block were similar in the two groups. The duration of sensory (242 +/- 76 versus 98 +/- 33 min) and motor (310 +/- 81 versus 130 +/- 31 min) blockade were significantly longer in the dexamethasone than in the control group (P < 0.01). We conclude that the addition of dexamethasone to lidocaine 1.5% solution in axillary brachial plexus block prolongs the duration of sensory and motor blockade. PMID- 16368841 TI - Continuous positive airway pressure breathing increases the spread of sensory blockade after low-thoracic epidural injection of lidocaine. AB - Factors affecting the distribution of sensory blockade after epidural injection of local anesthetics remain incompletely clarified. To evaluate if increasing intrathoracic pressure affects the spread of thoracic epidural anesthesia, we randomized 20 patients who received an epidural catheter at the T7-8 or T8-9 intervertebral space into 2 groups. The control group (n = 10) received an epidural test dose of 4 mL lidocaine 2% during spontaneous breathing at ambient pressure. The continuous positive airway pressure (n = 10) group received the same epidural test dose but during spontaneous respiration with 7.5 cm H2O continuous positive airway pressure. The groups were comparable with respect to demographic variables. Fifteen minutes after the conclusion of the epidural injection, the sensory block ranged from from T4 [median, interquartile range 2.75 segments] to T11 (interquartile range 3.5 segments) in the control group and from T5 (interquartile range 2.25 segments) to L2 (IQR 2.25 segments) in the continuous positive airway pressure group (P = 0.005 for the caudal border). The total number of segments blocked was 7 (median, interquartile range 2.25) in the control group and 11 (interquartile range 3.5) in the continuous positive airway pressure group (P = 0.004). The number of segments blocked caudad to the injection site was 3 (median, interquartile range 3.5) in the control group and 6 (interquartile range 2.25) in the continuous positive airway pressure group (P = 0.005). We conclude that continuous positive airway pressure increases the spread of sensory blockade in thoracic epidural anesthesia, primarily by a more caudad extension of sensory blockade. PMID- 16368842 TI - The influence of hyperbaric bupivacaine temperature on the spread of spinal anesthesia. AB - The distribution of spinal anesthesia is affected by the density and viscosity of the local anesthetic solution that, in turn, may be influenced by the temperature of the injectate. Our hypothesis in the present study was that the temperature of the injectate influences its distribution into the subarachnoid space. We measured the density and viscosity of hyperbaric 0.5% bupivacaine at 25 degrees C and 37 degrees C and tested the onset and extent of spinal anesthesia achieved by these solutions in 36 patients. The densities of the two solutions were similar (mean [sd]): 25 degrees C, 1.028 [0.000], versus 37 degrees C, 1.028 [0.000] (g/mL), but the viscosity was more at 25 degrees C than at 37 degrees C (0.01116 [0.00003] versus 0.00843 [0.00002] g x cm(-1) x s(-1); P < 0.001). The maximum cephalad extent of loss of pinprick sensation was significantly higher with 37 degrees C (T2 with 37 degrees C versus T5 with 25 degrees C; P < 0.001), but the time to achieve peak block height was similar. In conclusion, we showed a consistent, but modest, increase in the cephalad level of spinal anesthesia by warming hyperbaric bupivacaine 0.5% from 25 degrees C to 37 degrees C. Viscosity was reduced in the warmed solution, but it is unclear if this or other factors led to the difference in spinal anesthetic level. PMID- 16368843 TI - The effect of age on the systemic absorption and systemic disposition of ropivacaine after epidural administration. AB - Knowledge about the systemic absorption and disposition of ropivacaine after epidural administration is important in regard to its clinical profile and the risk of systemic toxicity. We investigated the influence of age on the pharmacokinetics of ropivacaine 1.0% after epidural administration, using a stable-isotope method. Twenty-four patients were enrolled in 1 of 3 groups according to age (group 1: 18-40 yr; group 2: 41-60 yr; group 3: > or =61 yr). Patients received 150 mg ropivacaine hydrochloride epidurally. After 25 min, patients received 50 mL 0.44 mg/mL deuterium-labeled ropivacaine (D3-ropivacaine) IV. Arterial blood samples were collected up to 24 h after epidural administration. Total plasma concentrations of ropivacaine and D3-ropivacaine were determined using liquid chromatography mass spectrometry. In the oldest patients, elimination half-life was significantly longer (ratio of the geometric means 0.60; 95% confidence interval, 0.37-0.99) and clearance was significantly decreased (mean difference, 194 mL/min; 95% confidence interval, 18-370 mL/min) compared with the youngest patients. The systemic absorption was biphasic. Absorption kinetics for ropivacaine (fractions absorbed: (F1, F2) and half-lives: (t(1/2),a1), t(1/2),a2) during the fast and slow absorption process: 0.27 +/- 0.08 and 0.77 +/- 0.12, respectively; 10.7 +/- 5.2 min and 248 +/- 64 min, respectively) were in the same range as for other long-acting local anesthetics. F1 was on average 0.11 (95% confidence interval, 0.002-0.22) higher in the youngest compared with the middle age group. Observed age-dependent pharmacokinetic differences do not likely influence the risk of systemic toxicity in the elderly after a single epidural dose of ropivacaine. PMID- 16368844 TI - Doxepin by topical application and intrathecal route in rats. AB - The tricyclic antidepressant, doxepin, has been reported to be a potent local anesthetic in rat sciatic nerve blockade. We hypothesized that topical doxepin has significantly longer antinociception compared with control and intrathecally compared with bupivacaine. Solutions of 0.3 mL of doxepin at 50, 75, and 100 mM and control (only the vehicle solution) were applied as a patch to the shaved dorsal skin of rats. After a 2-h contact interval, the patch was removed, and the rats were tested by three sets of six pinpricks. Inhibition of withdrawal to pain and cutaneous trunci muscle reflex were graded. In the second investigation, 60 muL of doxepin at 10, 20, and 50 mM was injected through intrathecal catheters implanted in the lumbar region of rats, which were evaluated for motor function, proprioception, and nociception. Topical doxepin at concentrations of 75 mM and 100 mM was significantly more effective than control (P < 0.05). Complete recovery for the 100-mM concentration occurred at 60 h, although two of five rats demonstrated erythema and scarring. Intrathecally, 20 mM of doxepin was not significantly different for motor and proprioceptive function from 23 mM (0.75%) bupivacaine; however, neurotoxicity (defined as persistent neurological deficit) commenced at 50 mM. PMID- 16368845 TI - Partial sensory and motor deficit of ipsilateral lower limb after continuous interscalene brachial plexus block. AB - We describe a partial sensory and motor block of the ipsilateral lower limb after interscalene infusion. After and injection of 20 mL of ropivacaine through the needle, the catheter was advanced 5 cm, and an infusion of ropivacaine 0.2% 5 mL/h commenced. Six hours later, the patient reported a left sensory and motor hemisyndrome, which resolved after the infusion was discontinued. Cervical computed tomography showed the tip of the catheter close to the intervertebral foramen at the C7-T1 level and several intravertebral paramedullar air bubbles. We conclude that the neurological symptoms were caused by an injection of local anesthetic via an interscalene catheter placed in proximity to the epidural space. To avoid this complication, we recommend advancing the catheter no more than 2-3 cm and performing frequent neurological evaluation of patients. PMID- 16368846 TI - Gender and recovery after general anesthesia combined with neuromuscular blocking drugs. AB - Previous studies suggest that women recover faster from general anesthesia than men, but it is unclear whether this is a result of a gender effect or differences in the pattern of drug administration or type of surgery. We performed a subset analysis comparing recovery characteristics from general anesthesia combined with neuromuscular blocking drugs of female and male patients, at risk of awareness, enrolled in a large trial testing the effectiveness of bispectral index (BIS) monitoring. We used multivariate statistical methods to adjust for differences in baseline characteristics, duration and extent of surgery, and anesthetic drug administration in 1079 patients (584 male, 495 female). Female patients had higher BIS values than male patients despite similar amounts of anesthetic drug administration (time-averaged mean [sd] BIS: male 44.6 [7.1] versus female 46.4 [6.6]; P = 0.005). Time to eye opening after anesthesia and time to eligibility to discharge from the postanesthesia care unit were less in women than men (male 13.9 [13.2] min versus female 10.6 [11.6] min; P < 0.001; male 133 [209] min versus female 78 [106] min; P < 0.001, respectively). These differences persisted after multivariate adjustment (both P < or = 0.001). Gender has an independent effect on recovery times in patients undergoing general anesthesia combined with neuromuscular blocking drugs, with women recovering faster than men. Higher BIS values during maintenance of anesthesia in women, despite similar amounts of anesthetic drug administration, suggests that women are less sensitive to the hypnotic effect of anesthetic drugs than men and may help explain faster recovery times in women. PMID- 16368847 TI - The effects of the alveolar recruitment maneuver and positive end-expiratory pressure on arterial oxygenation during laparoscopic bariatric surgery. AB - Abnormalities in gas exchange that occur during anesthesia are mostly caused by atelectasis, and these alterations are more pronounced in morbidly obese than in normal weight subjects. Sustained lung insufflation is capable of recruiting the collapsed areas and improving oxygenation in healthy patients of normal weight. We tested the effect of this ventilatory strategy on arterial oxygenation (Pao2) in patients undergoing laparoscopic bariatric surgery. After pneumoperitoneum was accomplished, the recruitment group received up to 4 sustained lung inflations with peak inspiratory pressures up to 50 cm H2O, which was followed by ventilation with 12 cm H2O positive end-expiratory pressure (PEEP). The patient's lungs in the control group were ventilated in a standard fashion with PEEP of 4 cm H2O. Variables related to gas exchange, respiratory mechanics, and hemodynamics were compared between recruitment and control groups. We found that alveolar recruitment effectively increased intraoperative Pao2 and temporarily increased respiratory system dynamic compliance (both P < 0.01). The effects of alveolar recruitment on oxygenation lasted as long as the trachea was intubated, and lungs were ventilated with high PEEP, but soon after tracheal extubation, all the beneficial effects on oxygenation disappeared. The mean number of vasopressor treatments given during surgery was larger in the recruitment group compared with the control group (3.0 versus 0.8; P = 0.04). In conclusion, our data suggest that the use of alveolar recruitment may be an effective mode of improving intraoperative oxygenation in morbidly obese patients. Our results showed the effect to be short lived and associated with more frequent intraoperative use of vasopressors. PMID- 16368848 TI - Does the timing of tracheal intubation based on neuromuscular monitoring decrease laryngeal injury? A randomized, prospective, controlled trial. AB - Vocal cord injuries (VCI) and postoperative hoarseness (PH) are common complications after general anesthesia. Poor muscle relaxation at the moment of tracheal intubation may result in VCI. There is a large interindividual variation in neuromuscular depression after administration of neuromuscular blocking drugs. Therefore, the optimal individual timing of tracheal intubation based on neuromuscular monitoring (monitoring) may decrease VCI. In this prospective trial, 60 patients were randomized into 2 groups: Monitoring group: tracheal intubation at maximum block based on monitoring after atracurium 0.5 mg/kg and 2 min group: tracheal intubation 2 min after injection of atracurium 0.5 mg/kg. Intubating conditions were evaluated with the Copenhagen score. VCI were examined by stroboscopy before and 24 and 72 h after surgery. PH was assessed at 24, 48, and 72 h after surgery by a standardized interview. Excellent intubating conditions were significantly increased in the monitoring group compared with the 2-min group: 8 versus 2 patients, respectively (P = 0.036). The incidence of PH between the study groups was comparable: 7 (monitoring) versus 8 patients (2-min) (P = 0.860). Similar findings were observed for VCI: 9 versus 5 patients; respectively (P = 0.268); type of VCI: thickening of the vocal cords: 8 (monitoring) versus 5 (2-min) patients (P = 0.423), hematomas: 2 patients in each group (not significant). The present study demonstrated that neuromuscular monitoring improved endotracheal intubating conditions. However, tracheal intubation at maximum intensity of neuromuscular block was not associated with a decrease in vocal cord injuries. PMID- 16368849 TI - Women with red hair report a slightly increased rate of bruising but have normal coagulation tests. AB - There is an anecdotal impression that redheads experience more perioperative bleeding complications than do people with other hair colors. We, therefore, tested the hypothesis that perceived problems with hemostasis could be detected with commonly used coagulation tests. We studied healthy female Caucasian volunteers, 18 to 40 yr of age, comparable in terms of height, weight, and age, with natural bright red (n = 25) or black or dark brown (n = 26) hair. Volunteers were questioned about their bleeding history and the following tests were performed: complete blood count, prothrombin time/international normalized ratio, activated partial thromboplastin time, platelet function analysis, and platelet aggregation using standard turbidimetric methodology. Agonists for aggregation were adenosine diphosphate, arachidonic acid, collagen, epinephrine, and two concentrations of ristocetin. The red-haired volunteers reported significantly more bruising, but there were no significant differences between the red-haired and dark-haired groups in hemoglobin concentration, platelet numbers, prothrombin time/international normalized ratio, or activated partial thromboplastin time. Furthermore, no significant differences in platelet function, as measured by platelet function analysis or platelet aggregometry, were observed. We conclude that if redheads have hemostasis abnormalities, they are subtle. PMID- 16368850 TI - Middle ear pressure changes during anesthesia with or without nitrous oxide are similar among airway devices. AB - We tested the hypothesis that middle ear pressure (MEP) is influenced by the choice of airway device during anesthesia with or without nitrous oxide (N2O) in the gas mixture. Eighty consecutive anesthetized, paralyzed ventilated patients (ASA physical status I-II, 18-65 yr) were randomly allocated for airway management with the orally inserted tracheal tube, classic laryngeal mask airway, ProSeal laryngeal mask airway, or laryngeal tube suction with or without N2O 66% in the gas mixture. MEP was measured from both ears in random order by a blinded observer before induction of anesthesia and every 10 min for 70 min. In the N2O groups, N2O was changed to air after 40 min. There were no differences in MEP among the airway devices in the N2O or air groups. MEP was unchanged in the air groups but increased in the N2O groups with N2O (P < 0.0001) and decreased with air (P < 0.02). Baseline values for MEP were similar, but MEP was always higher for the N2O groups (P < 0.001). We conclude that the choice of airway device does not influence MEP among orally inserted tracheal tube, classic laryngeal mask airway, ProSeal laryngeal mask airway, and laryngeal tube suction during anesthesia with or without N2O in the gas mixture. PMID- 16368851 TI - Severe postextubation laryngeal obstruction: the role of prior neck dissection and radiation. AB - Aggressive treatment of advanced head and neck cancer may result in more patients undergoing surgery for unrelated illnesses. We present a case of a patient requiring emergency tracheostomy placement after a routine liver segment resection 10 yr after undergoing a bilateral modified radical neck dissection and radiation therapy. This type of patient may be at increased risk for both postoperative laryngeal edema and neuronal imbalance secondary to their preoperative condition. PMID- 16368852 TI - Bradycardia and asystole during neuraxial anesthesia. PMID- 16368853 TI - Is local anesthetic overdose the most common cause of neuraxial cardiac arrest? PMID- 16368854 TI - Cardiac arrest during neuraxial anesthesia: are all databases comparable? PMID- 16368855 TI - Possible air embolism during eye surgery. PMID- 16368856 TI - Smoke in the operating room complex: a rare incident of internal disaster. PMID- 16368857 TI - Intraoperative heparin and heparin-induced thrombocytopenia. PMID- 16368858 TI - Introducer catheter for fiberoptic intubation does exist. PMID- 16368859 TI - Permanent loss of cervical spinal cord function associated with the posterior approach. PMID- 16368860 TI - GE Healthcare response to dangerous design flaw in the Ohmeda Aespire anesthesia system. PMID- 16368861 TI - Could an epidural hematoma after neuraxial blockade really be idiopathic? PMID- 16368862 TI - Remifentanil patient-controlled analgesia in labor. PMID- 16368863 TI - A comparison of the Vasotrac with invasive arterial blood pressure monitoring. PMID- 16368864 TI - TEG and perioperative hypercoagulability. PMID- 16368865 TI - Continuous anesthetic insufflation and topical anesthesia of the airway using Trachlight in chronic facial burns. PMID- 16368866 TI - Optimizing first-line treatment options for patients with advanced NSCLC. AB - The median survival time for advanced non-small cell lung cancer (NSCLC) remains poor, despite years of research into new chemotherapy combinations. Platinum based chemotherapy has long been the standard of care for the initial treatment of advanced NSCLC. While no one particular platinum-based chemotherapy regimen is definitely superior to the others (as demonstrated in the Eastern Cooperative Oncology Group's E1594 trial), three randomized phase III trials (the Southwest Oncology Group 9509, Italian Lung Cancer Project, and TAX326 trials) have recently demonstrated that taxane-platinum doublets are better tolerated than a combination of vinorelbine and cisplatin (VC). Moreover, a combination of docetaxel and cisplatin produced superior survival and quality of life than VC in the TAX326 study. Nonplatinum combinations, such as a taxane-gemcitabine doublet, appear promising and better tolerated than their platinum-based comparators in other studies. Efforts to evaluate chemotherapy specifically in elderly patients and in those with poor performance status (PS) have increased. Single-agent chemotherapy has been safely administered to these populations, but platinum based doublet therapy may also be feasible in both elderly patients and patients with PS scores of 2. The addition of the monoclonal antibody against vascular endothelial growth factor, bevacizumab, to standard chemotherapy for patients with non-squamous cell advanced NSCLC significantly extended median survival in the E4599 randomized trial. Each incremental advance demonstrates that progress can be made in first-line treatment of advanced NSCLC. PMID- 16368867 TI - Paradigm shift in the treatment of head and neck cancer: the role of neoadjuvant chemotherapy. AB - Chemotherapy is an integral component of the management of patients with locally advanced head and neck cancer, though the optimal use of chemotherapy remains to be defined. The combination of a platinum agent and 5-fluorouracil has been used as the standard neoadjuvant treatment and has been shown to permit organ preservation in operable patients and improve long-term survival outcomes in operable and inoperable patients. Recently, the addition of a taxane, docetaxel or paclitaxel, to standard platinum plus 5-fluorouracil induction chemotherapy has been shown to further improve response rates and survival outcomes. Phase III data are emerging to support combinations of docetaxel or paclitaxel with a platinum plus 5-fluorouracil as a new, more effective and less toxic standard for neoadjuvant chemotherapy. Sequential treatment regimens, incorporating a combination of induction chemotherapy and chemoradiation, are also under study in efforts to further improve long-term survival outcomes. Induction regimens incorporating docetaxel or paclitaxel with a platinum plus 5-fluorouracil are under evaluation in this setting. Randomized trials comparing a sequential treatment approach with standard therapies are also being undertaken and will likely define a new treatment paradigm for patients with locally advanced head and neck cancer. PMID- 16368868 TI - Extending survival with chemotherapy in metastatic breast cancer. AB - Metastatic breast cancer (MBC) remains essentially incurable, and goals of therapy include the palliation of symptoms, delay of disease progression, and prolongation of overall survival time without negatively impacting quality of life. Anthracycline and taxane-based therapies have traditionally shown the highest degree of activity in MBC. Though numerous randomized clinical trials have shown improvements in overall response rates, few have found clear survival benefits. In recent years, however, there has been a small but growing series of clinical trials demonstrating modest, but meaningful survival advantages in metastatic disease. A common feature in many of these trials has been the use of a taxane, and more recently, a taxane combined with an antimetabolite. In addition, the development of targeted biologic agents active against MBC, such as trastuzumab and bevacizumab, has demonstrated great potential for enhancing the effects of chemotherapy and producing meaningful survival improvements. The role of the taxanes, antimetabolites, and biologics in extending survival in MBC is discussed. PMID- 16368869 TI - Achieving treatment goals for hormone-refractory prostate cancer with chemotherapy. AB - The belief that hormone-refractory prostate cancer (HRPC) is a chemotherapy resistant disease has been effectively refuted by the results of two recent randomized phase III trials. The TAX327 trial compared weekly docetaxel, every-3 weeks (Q3W) docetaxel, and Q3W mitoxantrone plus prednisone in 1,006 patients with HRPC, and results demonstrated that survival was significantly longer with a docetaxel-based regimen than with mitoxantrone. That trial demonstrated that only Q3W docetaxel was significantly superior to mitoxantrone with respect to overall survival. Quality of life was also superior in the docetaxel groups. In the Southwest Oncology Group (SWOG) 9916 trial, 674 men with progressive HRPC were randomized to 3-week cycles of docetaxel plus estramustine or mitoxantrone plus prednisone. Overall and disease-free survival times were significantly longer in the docetaxel arm. Collectively, the results of these trials demonstrate that survival can be significantly improved with chemotherapy in patients with HRPC to an extent that is comparable with the survival benefits seen in other cancers considered sensitive to chemotherapy such as breast cancer. Among various research tasks in HRPC is the definition of potential surrogate end points for survival, which will facilitate the conduct of pivotal trials. Prostate-specific antigen (PSA) response rate and changes in PSA constructs (i.e., PSA doubling time and PSA velocity) are promising potential surrogate end points for future trials and are being actively evaluated at the present time. Until there is clear demonstration of a surrogate role for these alternative end points, survival remains the appropriate end point for phase III trials in HRPC. There is a need for safe and effective second- and third-line regimens for patients progressing after docetaxel, and these patients should enter clinical trials designed for this population. Mitoxantrone, vinorelbine, the platinum analogue satraplatin, and epothilone are among compounds that require careful testing in this setting. The addition of targeted therapies, such as the endothelin receptor antagonist, atrasentan, and angiogenesis inhibitors, such as thalidomide and bevacizumab, to docetaxel-based therapy is being evaluated. High-dose calcitriol may also be an effective addition to docetaxel. The extensive effort devoted to the evaluation of chemotherapy and other systemic modalities of treatment of HRPC is likely to yield additional clinical benefit for patients, making HRPC a more manageable, less lethal, and less debilitating disease. PMID- 16368870 TI - Advances in the treatment of metastatic colorectal cancer. AB - The overall 5-year survival rate for patients with metastatic colorectal cancer (CRC) is less than 10%. Median survival with 5-fluorouracil (5-FU)/leucovorin (LV) therapy is approximately 12 months. Recent additions to the chemotherapy armamentarium for this disease have begun to prolong median survival times. In trials in which patients are exposed to all three approved chemotherapy agents, oxaliplatin, irinotecan, and 5-FU/LV, or capecitabine during the course of their disease, median survival has reached 20 months. The addition of oxaliplatin and irinotecan to 5-FU/LV regimens has also led to the maintenance of quality of life for longer intervals than were traditionally observed with 5-FU/LV alone. Current standard first-line regimens for metastatic CRC are FOLFOX (infusional 5-FU/LV with oxaliplatin) and FOLFIRI (infusional 5-FU/LV with irinotecan). The addition of bevacizumab to a two-drug regimen (irinotecan with 5-FU/LV) prolongs median survival to 20 months, with a modest amount of additional toxicity. Improvements in this median survival have not yet been realized with modifications to the current standard regimens; however, the oral agent capecitabine appears to be a reasonable substitute for infusional 5-FU/LV in combination regimens or as a single agent, with the advantage of reducing the inconvenience of the long infusion time. Ongoing investigations will identify a place for capecitabine, epidermal growth factor inhibitors, and new cytotoxics in the treatment of metastatic CRC. PMID- 16368871 TI - Evolving chemotherapy for advanced gastric cancer. AB - Gastric cancer is the fourth most commonly diagnosed cancer and is the second leading cause of cancer death worldwide. More than 50% of patients undergo surgery, but even after a curative resection, 60% of patients relapse locally or with distant metastases. Despite the fact that many advances have occurred in the management of gastric cancer, it continues to carry a poor prognosis, amplifying the importance of palliative chemotherapy. When compared with best supportive care alone, combination chemotherapy yields a significant advantage in the management of advanced gastric cancer. However, no single regimen has emerged or been accepted as clearly superior over another. Numerous phase II studies have demonstrated promising results with newer agents including irinotecan, docetaxel, capecitabine, S-1, and oxaliplatin. Recently reported phase III results with these agents now demonstrate positive developments in the treatment options for patients with advanced gastric cancer. PMID- 16368872 TI - Genome-wide analysis of restriction-modification system in unicellular and filamentous cyanobacteria. AB - Cyanobacteria are an ancient group of gram-negative bacteria with strong genome size variation ranging from 1.6 to 9.1 Mb. Here, we first retrieved all the putative restriction-modification (RM) genes in the draft genome of Spirulina and then performed a range of comparative and bioinformatic analyses on RM genes from unicellular and filamentous cyanobacterial genomes. We have identified 6 gene clusters containing putative Type I RMs and 11 putative Type II RMs or the solitary methyltransferases (MTases). RT-PCR analysis reveals that 6 of 18 MTases are not expressed in Spirulina, whereas one hsdM gene, with a mutated cognate hsdS, was detected to be expressed. Our results indicate that the number of RM genes in filamentous cyanobacteria is significantly higher than in unicellular species, and this expansion of RM systems in filamentous cyanobacteria may be related to their wide range of ecological tolerance. Furthermore, a coevolutionary pattern is found between hsdM and hsdR, with a large number of site pairs positively or negatively correlated, indicating the functional importance of these pairing interactions between their tertiary structures. No evidence for positive selection is found for the majority of RMs, e.g., hsdM, hsdS, hsdR, and Type II restriction endonuclease gene families, while a group of MTases exhibit a remarkable signature of adaptive evolution. Sites and genes identified here to have been under positive selection would provide targets for further research on their structural and functional evaluations. PMID- 16368873 TI - Regulation of hypothalamic gene expression by glucocorticoid: implications for energy homeostasis. AB - The present study investigated the hypothalamic gene expressions regulated by glucocorticoids (GC), key hormones in energy homeostasis. Using the serial analysis of gene expression (SAGE) method, we studied the effects of adrenalectomy (ADX) and GC on the transcriptomes of mouse hypothalamus. Approximately 180,000 SAGE tags, which correspond to 50,000 tag species, were isolated from each group of intact or adrenalectomized mice as well as 1, 3, and 24 h after GC injection. ADX upregulated diazepam binding inhibitor gene expression while downregulating vomeronasal 1 receptor D4, genes involved in mitochondrial phosphorylation (cytochrome-c oxidase 1 and NADH dehydrogenase 3), 3beta-hydroxysteroid dehydrogenase-1, and prostaglandin D2 synthase. GC increased the gene expression levels of dehydrogenase/reductase member 3, prostaglandin D2 synthase, solute carrier family 4 member 4, and five cytoskeletal proteins including myosin light chain phosphorylatable fast and troponin C2 fast. On the other hand, GC reduced the mRNA levels of calmodulin 1 and expressed sequence tag similar to calmodulin 2, ATP synthase F0 subunit 6, and solute carrier family 4 member 3. Moreover, 7 uncharacterized and 43 novel transcripts were modulated by ADX and GC. The present study has identified genes that may regulate hypothalamic systems governing energy balance in response to ADX and GC. PMID- 16368874 TI - Gene expression profiling of diaphragm muscle in alpha2-laminin (merosin) deficient dy/dy dystrophic mice. AB - Deficiency of alpha2-laminin (merosin) underlies classical congenital muscular dystrophy in humans and dy/dy muscular dystrophy in mice and causes severe muscle dysfunction in both species. To gain greater insight into the biochemical and molecular events that link alpha2-laminin deficiency with muscle fiber necrosis, and the associated compensatory responses, gene expression profiles were characterized in diaphragm muscle from 8-wk-old dy/dy mice using oligonucleotide microarrays. Compared with age-matched normal muscle, dystrophic diaphragm was characterized by predominantly augmented gene expression, irrespective of the fold-change threshold. Among the 69 genes with at least plus or minus twofold significantly altered expression, 30 belonged to statistically overrepresented Gene Ontology (GO) biological process groups. These covered four specific themes: development including muscle development, cell motility with an emphasis on muscle contraction, defense/immune response, and cell adhesion. An additional 11 gene transcripts were assigned to more general overrepresented GO biological process groups (e.g., cellular process, organismal physiological process); the remaining 28 did not belong to any overrepresented groups. GO cellular constituent assignment resulted in the highest degree of overrepresentation in extracellular and muscle fiber locations, whereas GO molecular function assignment was most notable for various types of binding. RT-PCR was performed on 38 of 41 genes with at least plus or minus twofold significantly altered expression that were assigned to overrepresented GO biological process groups, with expression changes verified for 36 of 38 genes. These results indicate that several specific groups of genes have altered expression in response to genetic alpha2-laminin deficiency, with both similarities and differences compared with data reported for dystrophin-deficient muscular dystrophies. PMID- 16368875 TI - Distinct gene expression profiles in adult mouse heart following targeted MAP kinase activation. AB - Three major MAP kinase signaling cascades, ERK, p38, and JNK, play significant roles in the development of cardiac hypertrophy and heart failure in response to external stress and neural/hormonal stimuli. To study the specific function of each MAP kinase branch in adult heart, we have generated three transgenic mouse models with cardiac-specific and temporally regulated expression of activated mutants of Ras, MAP kinase kinase (MKK)3, and MKK7, which are selective upstream activators for ERK, p38, and JNK, respectively. Gene expression profiles in transgenic adult hearts were determined using cDNA microarrays at both early (4-7 days) and late (2-4 wk) time points following transgene induction. From this study, we revealed common changes in gene expression among the three models, particularly involving extracellular matrix remodeling. However, distinct expression patterns characteristic for each pathway were also identified in cell signaling, growth, and physiology. In addition, genes with dynamic expression differences between early vs. late stages illustrated primary vs. secondary changes on MAP kinase activation in adult hearts. These results provide an overview to both short-term and long-term effects of MAP kinase activation in heart and support some common as well as unique roles for each MAP kinase cascade in the development of heart failure. PMID- 16368876 TI - WTC deafness Kyoto (dfk): a rat model for extensive investigations of Kcnq1 functions. AB - KCNQ1 forms K+ channels by assembly with regulatory subunit KCNE proteins and plays a key role in the K+ homeostasis in a variety of tissues. In the heart, KCNQ1 is coassembled with KCNE1 to produce a cardiac delayed rectifier K+ current. In the inner ear, the KCNQ1/KCNE1 complex maintains the high concentration of K+ in the endolymph. In the stomach, KCNQ1 is coassembled with KCNE2 to form the K+ exflux channel that is essential for gastric acid secretion. In the colon and small intestine, KCNQ1 is coassembled with KCNE3 to play an important role in transepithelial cAMP-stimulated Cl- secretion. For further understanding of Kcnq1 function in vivo, an animal model has been required. Here we reported the identification of a coisogenic Kcnq1 mutant rat, named deafness Kyoto (dfk), and the characterization of its phenotypes. WTC-dfk rats carried intragenic deletion at the Kcnq1 gene and showed impaired gain of weight, deafness, and imbalance resulting from the marked reduction of endolymph, prolonged QT interval in the electrocardiogram (ECG), and gastric achlorhydria associated with hypertrophic gastric mucosa. Surprisingly, WTC-dfk rats showed hypertension, which suggested that Kcnq1 might be involved in the regulation of blood pressure. These findings suggest that WTC-dfk rats could represent a powerful tool for studying the physiological functions of KCNQ1 and for the establishment of new therapeutic procedures for Kcnq1-related diseases. PMID- 16368877 TI - Genomic annotation of 15,809 ESTs identified from pooled early gestation human eyes. AB - To complement cDNA libraries from the human eye at early gestation and to discover candidate genes associated with early ocular development, we used freshly dissected human eyeballs from week 9-14 of gestation to construct the early human fetal eye cDNA library. A total of 15,809 clones were isolated and sequenced from the unamplified and unnormalized library. We screened 11,246 good quality ESTs, leading to the identification of 5,534 nonredundant clusters. Among them, 4,010 (72%) genes matched in the human protein database (Ensembl). The remaining 28% (1,524) corresponded to potentially novel or previously unidentified ESTs. We used BLASTX to compare our EST data with eight organisms and found common expression of a high portion of genes: Caenorhabditis briggsae (26%), Caenorhabditis elegans (27%), Anopheles gambiae (37%), Drosophila melanogaster (32%), Danio rerio (42%), Fugu rubripes (49%), Rattus norvegicusvalitus (52%), and Mus musculus (59%). Nevertheless, 48% (2,680 of 5,534) of the genes expressed in the early developing eye were not shared with current NEIBank human eye cDNA data. In addition, eight known retinal disease genes existed in our ESTs. Among them, six (COL11A1, BBS5, PDE6B, OAT, VMD2, and PGK1) were conserved among the genomes of other organisms, indicating that our annotated EST set provides not only a valuable resource for gene discovery and functional genomic analysis but also for phylogenetic analysis. Our foremost early gestation human eye cDNA library could provide detailed comparisons across species to identify physiological functions of genes and to elucidate evolutionary mechanisms. PMID- 16368878 TI - Expansion of CD1d-restricted NKT cells in patients with primary HIV-1 infection treated with interleukin-2. AB - Innate CD1d-restricted natural killer T (NKT) cells are infected and lost in HIV 1-infected patients, and this could contribute to HIV-1 pathogenesis because NKT cells play an important role in directing both adaptive and innate immunity. Administration of interleukin-2 (IL-2) to HIV-1-infected patients leads to substantial and sustained CD4+ T-cell expansion, involving both naive and memory cells. We investigated whether IL-2 treatment could restore the NKT cell compartment in patients with primary HIV-1 infection. We show that IL-2 combined with effective antiretroviral therapy (ART) resulted in significant expansion of CD1d-restricted NKT cells. Expansion occurred in both the CD4- and CD4+ subsets of NKT cells, and expanded cells expressed the CD161 maturation marker while expression of the HIV coreceptor CCR5 was reduced. These data indicate that IL-2 treatment in combination with effective ART is beneficial for the restoration of innate NKT cell immunity in patients with primary HIV-1 infection. PMID- 16368879 TI - A DNA-based cancer vaccine enhances lymphocyte cross talk by engaging the NKG2D receptor. AB - The NKG2D receptor is a stimulatory receptor expressed on NK cells and activated CD8 T cells. We previously demonstrated that engaging the NKG2D receptor markedly improved the efficacy of a survivin-based DNA vaccine. The combination vaccine, encoding both the NKG2D ligand H60 and survivin, activates innate and adaptive antitumor immunity and results in better protection against tumors of different origin and NKG2D expression levels. Here we demonstrate that the enhanced vaccine efficacy is in part attributable to increased cross talk between lymphocytes. Depletion of CD8 T cells during priming reduces the vaccine-induced activation of dendritic cells (DCs) and NK cell activity. Depletion of NK cells during priming leads to reduced DC activation and CTL activity. However, depletion of CD4 T cells results in the activation of DCs, NK cells, and CD8 T cells and enhances NK cell activity. The pH60/Survivin vaccine also increases DCs and NK cells but decreases CD4 T cell homing to Peyer patches, presumably as a result of changes in the homing receptor profile. Thus, by preferentially activating and attracting positive regulators and reducing negative regulators in Peyer patches, this dual function DNA vaccine induces a microenvironment more suitable for NK cell activation and T cell priming. PMID- 16368880 TI - Coexistence of multiple mechanisms of PT523 resistance in human leukemia cells harboring 3 reduced folate carrier alleles: transcriptional silencing, inactivating mutations, and allele loss. AB - The reduced folate carrier (RFC) is the dominant route for the uptake of various antifolates including PT523, a potent dihydrofolate reductase inhibitor (Ki = 0.35 pM) and an excellent transport substrate of the RFC (Kt = 0.7 microM). Here, we describe the multiple mechanisms of RFC inactivation in human leukemia PT523 resistant cells originally harboring 3 RFC alleles. Cellular exposure to gradually increasing PT523 concentrations resulted in sublines displaying up to 3500-fold resistance to various hydrophilic antifolates that rely on RFC for their cellular uptake. Antifolate-resistant cells lost RFC gene expression (65% 99% loss) due to impaired promoter binding of various transcription factors that regulate RFC gene expression. Additionally, DNA sequencing revealed that PT523 resistant cells contained a cluster of 4 nearly consecutive mutations residing on a single RFC allele including L143P, A147V, R148G, and Q150Stop. Southern blot analysis established the loss of an RFC allele in PT523-resistant cells. These alterations resulted in markedly decreased RFC protein levels (approximately 80% 99% loss) and consequently impaired [3H]methotrexate transport (87%-99% loss). This study provides the first evidence that acquisition of PT523 resistance in human leukemia cells harboring 3 RFC alleles is due to multiple coexisting alterations including transcriptional silencing, inactivating mutations, and RFC allele loss. PMID- 16368881 TI - Red blood cells express a functional endothelial nitric oxide synthase. AB - The synthesis of nitric oxide (NO) in the circulation has been attributed exclusively to the vascular endothelium. Red blood cells (RBCs) have been demonstrated to carry a nonfunctional NO synthase (NOS) and, due to their huge hemoglobin content, have been assumed to metabolize large quantities of NO. More recently, however, RBCs have been identified to reversibly bind, transport, and release NO within the cardiovascular system. We now provide evidence that RBCs from humans express an active and functional endothelial-type NOS (eNOS), which is localized in the plasma membrane and the cytoplasm of RBCs. This NOS is regulated by its substrate L-arginine, by calcium, and by phosphorylation via PI3 kinase. RBC-NOS activity regulates deformability of RBC membrane and inhibits activation of platelets. The NOS-dependent conversion of L-arginine in RBCs is comparable to that of cultured human endothelial cells. RBCs in eNOS-/- mice in contrast to wild-type mice lack NOS protein and activity, strengthening the evidence of an eNOS in RBCs. These data show an eNOS-like protein and activity in RBCs serving regulatory functions in RBCs and platelets, which may stimulate new approaches in the treatment of NO deficiency states inherent to several vascular and hematologic diseases. PMID- 16368882 TI - Extracorporeal photochemotherapy for the treatment of steroid-resistant chronic GVHD. AB - Chronic graft-versus-host disease (GVHD) is a major limitation of successful allogeneic hematopoietic stem cell transplantation (HSCT). Extracorporeal photochemotherapy (ECP) has been tested extensively in small cohorts of patients with chronic GVHD. In this study, we retrospectively evaluated 71 patients with severe chronic GVHD treated with ECP. Response rate was 61% (n = 43), and 14 patients had complete responses (CRs). The best responses were observed in skin, liver, oral mucosa, and eye. Factors affecting outcomes were assessed in the less heavily pretreated subgroup (n = 63). Thrombocytopenia was associated with a lower response rate (P = .04), and there was a trend toward a higher response rate in de novo chronic GVHD. At 6 months, a total of 27 (69%) of 39 patients who were alive continued to have a sustained response (CR 4 [10%] of 39, and partial response [PR] 23 [59%] of 39). The cumulative incidence of steroid discontinuation at 1 year was 22%. The overall survival since initiation of therapy was 53% at 1 year. Response to ECP and platelet count at initiation of therapy were the strongest predictors of nonrelapse mortality (NRM) on univariate analysis. Objective responses were observed in a substantial number of patients with both skin and visceral chronic GVHD failing corticosteroids and other immunosuppression. PMID- 16368884 TI - Frequent genomic alterations in epithelium measured by microsatellite instability following allogeneic hematopoietic cell transplantation in humans. AB - Although typically found in cancers, frameshift mutations in microsatellites have also been detected in chronically inflamed tissues. Allogeneic hematopoietic cell transplantation (HCT) may potentially produce chronic tissue stress through graft versus-host reactions. We examined non-neoplastic epithelial tissues (colon, buccal) obtained 1 to 5061 days after human allogeneic HCT for the presence of genomic alterations at 3 tetranucleotide and 3 mononucleotide microsatellite loci. Novel bands indicative of microsatellite instability (MSI) at tetranucleotide repeats were detected in laser-microdissected colonic crypts and in buccal smears of 75% and 42% of patients who received an allograft, respectively. In contrast, no MSI was found in similar tissues from control subjects and from patients after intensive chemotherapy or in buccal cells from patients after autologous HCT. The MSI found in colon, which was often affected by graft-versus-host disease, was not due to loss of expression or nitrosylation of DNA repair proteins. MSI in clinically intact oral mucosa was more frequently found at later time points after HCT. MSI was also found in 3 posttransplant squamous cell cancers examined. Our data show that genomic alterations in epithelium regularly occur after allogeneic HCT and may be implicated in the evolution of posttransplantation diseases, including secondary cancer. PMID- 16368883 TI - Size of FLT3 internal tandem duplication has prognostic significance in patients with acute myeloid leukemia. AB - FLT3 internal tandem duplications (FLT3/ITDs) in the juxtamembrane domain are found in approximately 25% of acute myeloid leukemia (AML) patients, ranging in size from 3 to hundreds of nucleotides. We examined whether the sizes of FLT3/ITDs were associated with clinical outcomes in 151 AML patients enrolled in Southwest Oncology Group studies: S9333 and S9500. FLT3/ITDs were identified in 32% of patients (median ITD size = 39 nucleotides; range, 15-153 nucleotides). The CR rates were 35%, 67%, and 52% for patients with large (>or= 40), small (< 40), and no ITDs, respectively (P = .19). Increasing ITD size was associated with decreasing OS (estimated 5-year OS: large = 13%, small = 26%, and no ITD = 21%, P = .072) and RFS (estimated 5-year RFS: large = 13%, small = 27%, and no ITD = 34%, P = .017). These studies suggest that ITD size may have prognostic significance. PMID- 16368885 TI - Identifying a common molecular mechanism for inhibition of MITF and STAT3 by PIAS3. AB - Protein inhibitor of activated STAT3 (PIAS3) functions in vivo as a key molecule in suppressing the transcriptional activity of both microphthalmia transcription factor (MITF) and signal transducer and activator of transcription 3 (STAT3), 2 transcription factors that play a major role in the regulation of growth and function in mast cells and melanocytes. Previously, we have demonstrated binding of PIAS3 to MITF leading to the inhibition of MITF transcriptional activity. Following cellular activation, PIAS3 is released from MITF and binds to STAT3. Now we have localized a common binding motif in PIAS3 for MITF and STAT3. This motif (PIAS82-132), which contains 50 amino acids, is sufficient for the inhibition of both MITF and STAT3. Three-dimensional protein modeling demonstrated that this motif contains 2 alpha helices. Disruption of one of the helices led to the loss of PIAS3 inhibitory activity. In addition to contributing to our understanding of the mechanisms of PIAS3 activity, these results could pave the way toward the formulation of an antioncogenic agent for the inhibition of both STAT3 and MITF. PMID- 16368886 TI - Thrombopoietin regulates IEX-1 gene expression through ERK-induced AML1 phosphorylation. AB - The extracellular signal-regulated kinases (ERKs) are required for thrombopoietin (TPO) functions on hematopoietic cells, but the ERKs targets involved remain unknown. Here we show that the regulation of the immediate early gene X-1 (IEX 1), identified as an ERK substrate in response to TPO, was mediated by an ERK dependent phosphorylation of AML1. The addition of TPO to UT7-Mpl cells and primary megakaryocytes induced gene expression of IEX-1. Neither erythropoietin (EPO) nor granulocyte macrophage-colony stimulating factor (GM-CSF) was able to activate IEX-1 gene expression in UT7-Mpl cells. The induced expression was mediated by a transcriptional activation of the IEX-1 promoter and required an AML1-binding site located at -1068. The direct involvement of AML1 in the regulation of IEX-1 gene expression was shown by both the use of AML1 mutants and by shRNA experiments targeting endogenous AML1. Finally, the ability of TPO to induce the IEX-1 gene expression was inhibited by U0126, a specific inhibitor of the ERKs activator MEK and AML1 transcriptional activity was shown to be modulated by TPO through ERK-dependent phosphorylation. Taken together, these data suggest that AML1 plays a role in modulating the IEX-1 expression and that the ERK-dependent AML1 phosphorylation regulates the TPO-mediated activation of IEX-1. PMID- 16368887 TI - Cross talk among Notch3, pre-TCR, and Tal1 in T-cell development and leukemogenesis. AB - Integrated pathways are believed to determine hematopoietic cell fate and/or neoplastic transformation. Notch signaling has been shown to regulate T-cell differentiation and leukemogenesis. However, specific target genes and molecular partners are not fully elucidated. We show that Notch3 activation sustains aberrant SCL/Tal1 overexpression and phosphorylation in mature thymocytes. Furthermore, we define the role of SCL/Tal1 as a component of an activator complex, including phosphorylated Tal1 and Sp1, that specifically enhances cyclin D1 expression and demonstrate that Tal1/Sp1 specifically co-occupy the D1 promoter in vivo, only in the presence of pre-T-cell receptor (TCR). We therefore conclude not only that cyclin D1 is a target of the Tal1/Sp1 complex, but also that Notch3-dependent activation of pre-TCR/ERK signaling regulates SCL/Tal1 function. PMID- 16368888 TI - Complete deficiency in ADAMTS13 is prothrombotic, but it alone is not sufficient to cause thrombotic thrombocytopenic purpura. AB - ADAMTS13 is a plasma metalloproteinase that regulates platelet adhesion and aggregation through cleavage of von Willebrand factor (VWF) multimers. In humans, genetic or acquired deficiency in ADAMTS13 causes thrombotic thrombocytopenic purpura (TTP), a condition characterized by thrombocytopenia and hemolytic anemia with microvascular platelet thrombi. In this study, we report characterization of mice bearing a targeted disruption of the Adamts13 gene. ADAMTS13-deficient mice were born in the expected mendelian distribution; homozygous mice were viable and fertile. Hematologic and histologic analyses failed to detect any evidence of thrombocytopenia, hemolytic anemia, or microvascular thrombosis. However, unusually large VWF multimers were observed in plasma of homozygotes. Thrombus formation on immobilized collagen under flow was significantly elevated in homozygotes in comparison with wild-type mice. Thrombocytopenia was more severely induced in homozygotes than in wild-type mice after intravenous injection of a mixture of collagen and epinephrine. Thus, a complete lack of ADAMTS13 in mice was a prothrombotic state, but it alone was not sufficient to cause TTP-like symptoms. The phenotypic differences of ADAMTS13 deficiencies between humans and mice may reflect differences in hemostatic system functioning in these species. Alternatively, factors in addition to ADAMTS13 deficiency may be necessary for development of TTP. PMID- 16368889 TI - U1 small nuclear ribonucleoprotein immune complexes induce type I interferon in plasmacytoid dendritic cells through TLR7. AB - Plasmacytoid dendritic cells (PDCs), which produce IFN-alpha in response to autoimmune complexes containing nuclear antigens, are thought to be critically involved in the pathogenesis of systemic lupus erythematosus (SLE). One of the immunostimulatory components of SLE immune complexes (SLE-ICs) is self DNA, which is recognized through Tlr9 in PDCs and B cells. Small nuclear ribonucleoproteins (snRNPs) are another major component of SLE-ICs in 30% to 40% of patients. In this study, we show that murine PDCs are activated by purified U1snRNP/anti-Sm ICs to produce IFN-alpha and proinflammatory cytokines and to up-regulate costimulatory molecules. The induction of IFN-alpha and IL-6 by U1snRNPs in murine bone marrow-derived PDCs required the presence of intact U1RNA and was largely dependent on Tlr7 but independent of Tlr3. Intracellularly delivered isolated U1snRNA and oligoribonucleotides derived from the stem loop regions and the Sm-binding site of U1snRNA efficiently induced IFN-alpha and IL-6 in Flt3L cultured DCs in a Tlr7-dependent manner. The U1snRNA component of U1snRNP immune complexes, found in patients with SLE, acts as an endogenous "self" ligand for Tlr7 and triggers IFN-alpha and IL-6 production in PDCs. PMID- 16368890 TI - Apoptosis and complement-mediated lysis of myeloma cells by polyclonal rabbit antithymocyte globulin. AB - Current monoclonal antibody therapies for multiple myeloma have had limited success, perhaps due to narrow target specificity. We have previously described the ability of polyclonal rabbit antithymocyte globulin (rATG) to induce caspase- and cathepsin-mediated apoptosis in human B and plasma cells. We now extend this observation to myeloma cells. Complement independent cell death was measured after addition of rATG (1-1000 microg/mL) to cultures of myeloma cell lines or primary CD138+ isolates from patient bone marrow aspirates. rATG induced significant levels of apoptosis in myeloma cells as assayed by caspase induction, annexin V binding, subdiploid DNA fragmentation, plasma-membrane permeability, and loss of mitochondrial-membrane potential. Addition of complement greatly augmented myeloma-cell death. Binding of rATG to individual myeloma cell-surface proteins, primarily CD38, CD52, CD126, and CD138, was demonstrated by competitive inhibition experiments with targeted monoclonal antibodies. Three pathways of cell death were identified involving caspase activation, cathepsin D, and the genistein sensitive tyrosine kinase pathway. Fab'2 fragments of rATG had reduced proapoptotic activity, which was restored by coincubation with Fc fragments, and anti-CD32 or anti-CD64 antibodies. We conclude that rATG is an effective agent for in vitro induction of apoptosis in multiple myeloma, and that exploratory clinical trials may be warranted. PMID- 16368891 TI - A transforming growth factor-beta-induced protein stimulates endocytosis and is up-regulated in immature dendritic cells. AB - Dendritic cells (DCs) exhibit distinct functional properties at immature and mature states. To identify genes preferentially regulated in monocyte-derived immature DCs (imDCs), 13 000-element microarrays were hybridized with RNA isolated from imDCs, mature DCs (mDCs), monocytes, and macrophages and a TGF-beta induced protein (betaig-h3) was identified as being most prominently up-regulated in imDCs. By polymerase chain reaction (PCR), little betaig-h3 mRNA was detected in monocytes and macrophages, but it was abundant in imDCs. On DC activation with LPS, betaig-h3 mRNA became diminished, and in tissues, betaig-h3 mRNA was abundantly expressed in lymphoid-rich tissues such as the spleen, bone marrow, small intestines, and colon. betaig-h3 was expressed in 293T cells and purified as a 70-kDa protein and, by Western blotting, betaig-h3 was predominantly detected in the medium of imDCs. We demonstrate that betaig-h3 binds to macrophages and imDCs but not to mDCs and activates the Rac GTPase in macrophages, stimulating macrophage membrane ruffling and enhancing macrophage endocytosis. imDC endocytosis was also inhibited by purified anti-betaig-h3 antibodies. Therefore, betaig-h3 appears to be selectively up-regulated in imDCs to regulate antigen uptake through endocytosis. PMID- 16368892 TI - Role of CXCR3 carboxyl terminus and third intracellular loop in receptor-mediated migration, adhesion and internalization in response to CXCL11. AB - The chemokine receptor CXCR3 is predominantly expressed on activated T and natural killer (NK) cells. CXCR3 and its ligands, CXCL11, CXCL10, and CXCL9, play a major role in T-helper 1 (Th1)-dependent inflammatory responses. CXCL11 is the most dominant physiological inducer of adhesion, migration, and internalization of CXCR3. To study the role of CXCR3 carboxyl-terminus and the third intracellular (3i) loop in chemokine-mediated migration, adhesion, and CXCR3 internalization, we generated CXCR3 receptors mutated in their distal (Ser-Thr domain) or proximal (trileucine domain) membrane carboxyl terminus, and/or the third intracellular loop. We found that migration of CXCR3-expressing HEK 293 cells toward CXCL11 was pertussis toxin-dependent and required the membrane proximal carboxyl terminus of CXCR3. Internalization induced by CXCL11 and protein kinase C (PKC) activation was also regulated by the membrane proximal carboxyl terminus; however, only CXCL11-induced internalization required the LLL motif of this region. Internalization and Ca(2+) flux induced by CXCL11 were independent of the 3i loop S245, whereas migration at high CXCL11 concentrations, integrin-dependent adhesion, and actin polymerization were S245 dependent. Our findings indicate that CXCL11-dependent CXCR3 internalization and cell migration are regulated by the CXCR3 membrane proximal carboxyl terminus, whereas adhesion is regulated by the 3i loop S245. Thus, distinct conformational changes induced by a given CXCR3 ligand trigger different downstream effectors of adhesion, motility, and CXCR3 desensitization. PMID- 16368893 TI - Endogenous regulator of G-protein signaling proteins regulate the kinetics of Galphaq/11-mediated modulation of ion channels in central nervous system neurons. AB - Slow synaptic potentials are generated when metabotropic G-protein-coupled receptors activate heterotrimeric G-proteins, which in turn modulate ion channels. Many neurons generate excitatory postsynaptic potentials mediated by G proteins of the Galphaq/11 family, which in turn activate phospholipase C-beta. Accessory GTPase-activating proteins (GAPs) are thought to be required to accelerate GTP hydrolysis and rapidly turn off G-proteins, but the involvement of GAPs in neuronal Galphaq/11 signaling has not been examined. Here, we show that regulator of G-protein signaling (RGS) proteins provide necessary GAP activity at neuronal Galphaq/11 subunits. We reconstituted inhibition of native 2-pore domain potassium channels in cerebellar granule neurons by expressing chimeric Galpha subunits that are activated by Galphai/o-coupled receptors, thus bypassing endogenous Galphaq/11 subunits. RGS-insensitive variants of these chimeras mediated inhibition of potassium channels that developed and recovered more slowly than inhibition mediated by RGS-sensitive (wild-type) chimeras or native Galphaq/11 subunits. These changes were not accompanied by a change in agonist sensitivity, as might be expected if RGS proteins acted primarily as effector antagonists. The slowed recovery from potassium channel inhibition was largely reversed by an additional mutation that mimics the RGS-bound state. These results suggest that endogenous RGS proteins regulate the kinetics of rapid Galphaq/11 mediated signals in central nervous system neurons by providing GAP activity. PMID- 16368894 TI - Lysophosphatidylserine stimulates L2071 mouse fibroblast chemotactic migration via a process involving pertussis toxin-sensitive trimeric G-proteins. AB - Lysophosphatidylserine (LPS) may be generated after phosphatidylserine-specific phospholipase A2 activation. However, the effects of LPS on cellular activities and the identities of its target molecules have not been fully elucidated. In this study, we observed that LPS stimulates an intracellular calcium increase in L2071 mouse fibroblast cells, and that this increase was inhibited by 1-[6 ((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione (U-73122) but not by pertussis toxin, suggesting that LPS stimulates calcium signaling via G-protein coupled receptor-mediated phospholipase C activation. Moreover, LPS-induced calcium mobilization was not inhibited by the lysophosphatidic acid receptor antagonist, (S)-phosphoric acid mono-{2-octadec-9 enoylamino-3-[4-(pyridine-2-ylmethoxy)-phenyl]-propyl} ester (VPC 32183), thus indicating that LPS binds to a receptor other than lysophosphatidic acid receptors. It was also found that LPS stimulates two types of mitogen-activated protein kinase [i.e., extracellular signal-regulated protein kinase (ERK) and p38 kinase] in L2071 cells. Furthermore, these LPS-induced ERK and p38 kinase activations were inhibited by pertussis toxin, which suggests the role of pertussis toxin-sensitive G-proteins in the process. In terms of functional issues, LPS stimulated L2071 cell chemotactic migration, which was completely inhibited by pertussis toxin, indicating the involvement of pertussis toxin sensitive G(i) protein(s). This chemotaxis of L2071 cells induced by LPS was also dramatically inhibited by 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and by 2'-amino-3'-methoxyflavone (PD98059). This study demonstrates that LPS stimulates at least two different signaling cascades, one of which involves a pertussis toxin-insensitive but phospholipase C-dependent intracellular calcium increase, and the other involves a pertussis toxin sensitive chemotactic migration mediated by phosphoinositide 3-kinase and ERK. PMID- 16368895 TI - Hydralazine inhibits rapid acrolein-induced protein oligomerization: role of aldehyde scavenging and adduct trapping in cross-link blocking and cytoprotection. AB - Hydralazine strongly suppresses the toxicity of acrolein, a reactive aldehyde that contributes to numerous health disorders. At least two mechanisms may underlie the cytoprotection, both of which involve the nucleophilic hydrazine possessed by hydralazine. Under the simplest scenario, hydralazine directly scavenges free acrolein, decreasing intracellular acrolein availability and thereby suppressing macromolecular adduction. In a second "adduct-trapping" mechanism, the drug forms hydrazones with acrolein-derived Michael adducts in cell proteins, preventing secondary reactions of adducted proteins that may trigger cell death. To identify the most important mechanism, we explored these two pathways in mouse hepatocytes poisoned with the acrolein precursor allyl alcohol. Intense concentration-dependent adduct-trapping in cell proteins accompanied the suppression of toxicity by hydralazine. However, protective concentrations of hydralazine did not alter extracellular free acrolein levels, cellular glutathione loss, or protein carbonylation, suggesting that the cytoprotection is not due to minimization of intracellular acrolein availability. To explore ways whereby adduct-trapping might confer cytoprotection, the effect of hydralazine on acrolein-induced protein cross-linking was examined. Using bovine pancreas ribonuclease A as a model protein, acrolein caused rapid time- and concentration-dependent cross-linking, with dimerized protein detectable within 45 min of commencing protein modification. Lysine adduction in monomeric protein preceded the appearance of oligomers, whereas reductive methylation of protein amine groups abolished both adduction and oligomerization. Hydralazine inhibited cross-linking if added 30 min after commencing acrolein exposure but was ineffective if added after a 90-min delay. Adduct-trapping closely accompanied the inhibition of cross-linking by hydralazine. These findings suggest that cross-link blocking may contribute to hydralazine cytoprotection. PMID- 16368896 TI - Metagenomics to paleogenomics: large-scale sequencing of mammoth DNA. AB - We sequenced 28 million base pairs of DNA in a metagenomics approach, using a woolly mammoth (Mammuthus primigenius) sample from Siberia. As a result of exceptional sample preservation and the use of a recently developed emulsion polymerase chain reaction and pyrosequencing technique, 13 million base pairs (45.4%) of the sequencing reads were identified as mammoth DNA. Sequence identity between our data and African elephant (Loxodonta africana) was 98.55%, consistent with a paleontologically based divergence date of 5 to 6 million years. The sample includes a surprisingly small diversity of environmental DNAs. The high percentage of endogenous DNA recoverable from this single mammoth would allow for completion of its genome, unleashing the field of paleogenomics. PMID- 16368897 TI - In vivo characterization of the novel imidazopyridine BYK191023 [2-[2-(4-methoxy pyridin-2-yl)-ethyl]-3H-imidazo[4,5-b]pyridine], a potent and highly selective inhibitor of inducible nitric-oxide synthase. AB - Excessive release of nitric oxide from inducible nitric-oxide synthase (iNOS) has been postulated to contribute to pathology in a number of inflammatory diseases. We recently identified imidazopyridine derivatives as a novel class of potent nitricoxide synthase inhibitors with high selectivity for the inducible isoform. In the present study, we tested the in vivo potency of BYK191023 [2-[2-(4-methoxy pyridin-2-yl)-ethyl]-3H-imidazo-[4,5-b]pyridine], a selected member of this inhibitor class, in three different rat models of lipopolysaccharide-induced systemic inflammation. Delayed administration of BYK191023 dose-dependently suppressed the lipopolysaccharide-induced increase in plasma nitrate/nitrite (NO(x)) levels with an ED(50) of 14.9 micromol/kg/h. In a model of systemic hypotension following high-dose lipopolysaccharide challenge, curative administration of BYK191023 at a dose that inhibited 83% of the NO(x) increase completely prevented the gradual decrease in mean arterial blood pressure observed in vehicle-treated control animals. The vasopressor effect was specific for endotoxemic animals since BYK191023 did not affect blood pressure in saline challenged controls. In addition, in a model of lipopolysaccharide-induced vascular hyporesponsiveness, BYK191023 infusion partially restored normal blood pressure responses to norepinephrine and sodium nitroprusside via an l-arginine competitive mechanism. Taken together, BYK191023 is a member of a novel class of highly isoform-selective iNOS inhibitors with promising in vivo activity suitable for mechanistic studies on the role of selective iNOS inhibition as well as clinical development. PMID- 16368898 TI - KN-93 (2-[N-(2-hydroxyethyl)]-N-(4-methoxybenzenesulfonyl)]amino-N-(4 chlorocinnamyl)-N-methylbenzylamine), a calcium/calmodulin-dependent protein kinase II inhibitor, is a direct extracellular blocker of voltage-gated potassium channels. AB - The effect of Ca(2+)/calmodulin-dependent protein kinase II (CaMK II) on voltage gated ion channels is widely studied through the use of specific CaMK II blockers such as 2-[N-(2-hydroxyethyl)]-N-(4methoxybenzenesulfonyl)]amino-N-(4 chlorocinnamyl)-N-methylbenzylamine (KN-93). The present study demonstrates that KN-93 is a direct extracellular blocker of a wide range of cloned Kv channels from a number of different subfamilies. In all channels tested, the effect of 1 microM KN-93 was independent of CaMK II because 1 microM2-[N-(4 methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine, phosphate (KN-92), an inactive analog of KN-93, caused similar inhibition of currents. In addition, dialysis of cells with 10 microM CaMK II inhibitory peptide fragment 281-301 (CIP) had no effect on current kinetics and did not prevent the inhibitory effect of KN-93. The IC(50) for block of the Kv1.5 channel (used as an example to determine the nature of KN-93 block) was 307 +/- 12 nM. KN-93 blocked open channels with little voltage dependence that did not alter the V(1/2) of channel activation. Removal of P/C-type inactivation by mutation of arginine 487 to valine in the outer pore region of Kv1.5 (R487V) greatly reduced KN-93 block, whereas enhancement of inactivation induced by mutation of threonine 462 to cysteine (T462C) increased the potency of KN-93 by 4-fold. This suggested that KN 93 acted through promotion and stabilization of C-type inactivation. Importantly, KN-93 was ineffective as a blocker when applied intracellularly, suggesting that CaMK II-independent effects of KN-93 on Kv channels can be circumvented by intracellular application of KN-93. PMID- 16368899 TI - Discovery of a dual-function peptide that combines aminopeptidase N inhibition and kinin B1 receptor antagonism. AB - Previous analyses support that aminopeptidase N is a major inactivation pathway for high-affinity peptide ligands of the human and rabbit forms of the kinin B(1) receptor (agonists or antagonists). In this study, we found that the high affinity antagonist B-9958 (Lys-Lys-[Hyp(3), CpG(5), D-Tic(7), CpG(8)]des-Arg(9) BK; des-Arg(9)-BK, des-arginine(9)-bradykinin) is an aminopeptidase N substrate based on its capacity to compete for the hydrolysis of the chromogenic substrate L-Ala-p-nitroanilide by membranes isolated from human or rabbit arterial smooth muscle cells, its inactivation in the presence of these membranes (radioreceptor assay) and on its intense potentiation by the aminopeptidase N inhibitor amastatin in the rabbit aorta contractility assay (gain of 0.84 units in the pA(2) scale). Analogs of B-9958 in which the N-terminal Lys residue was substituted by D-Lys or D-Arg (B-10352 and B-10356, respectively) showed reduced affinity at the human or rabbit B(1) receptors (1.2-2.8-fold), as estimated by the displacement of [(3)H]Lys-des-Arg(9)-BK binding, but were more potent antagonists of des-Arg(9)-BK-induced contraction of the rabbit aorta than B-9958 in the absence of amastatin; they were not potentiated by the latter inhibitor. Unexpectedly, B-10356 inhibited L-Ala- p-nitroanilide hydrolysis without being inactivated, suggesting that it is an aminopeptidase N inhibitor. This was verified because B-10356 (but not B-10352) potentiated peptides unrelated to kinins but susceptible to aminopeptidase N inactivation (angiotensin III, thrombin receptor hexapeptide agonist). B-10356 inhibits dual molecular targets (aminopeptidase N enzyme K(i), 0.9-2.2 microM; kinin B(1) receptor binding K(i), 0.5-1.5 nM), and this may be an advantage for specific therapeutic applications (e.g., inhibition of angiogenesis). PMID- 16368900 TI - Inhibition of B16 melanoma metastases with the ruthenium complex imidazolium trans-imidazoledimethylsulfoxide-tetrachlororuthenate and down-regulation of tumor cell invasion. AB - The antimetastatic ruthenium complex imidazolium trans-imidazoledimethylsulfoxide tetrachlorouthenate (NAMI-A) is tested in the B16 melanoma model in vitro and in vivo. Treatment of B6D2F1 mice carrying intra-footpad B16 melanoma with 35 mg/kg/day NAMI-A for 6 days reduces metastasis weight independently of whether NAMI-A is given before or after surgical removal of the primary tumor. Metastasis reduction is unrelated to NAMI-A concentration, which is 10-fold lower than on primary site (1 versus 0.1 mM), and is correlated to the reduction of plasma gelatinolitic activity and to the decrease of cells expressing CD44, CD54, and integrin-beta(3) adhesion molecules. Metastatic cells also show the reduction of the S-phase cells with accumulation in the G(0)/G(1) phase. In vitro, on the highly metastatic B16F10 cell line, NAMI-A reduces cell Matrigel invasion and its ability to cross a layer of endothelial cells after short exposure (1 h) to 1 to 100 microM concentrations. In these conditions, NAMI-A reduces the gelatinase activity of tumor cells, and it also increases cell adhesion to poly-L-lysine and, in particular, to fibronectin, and this effect is associated to the increase of F-actin condensation. This work shows the selective effectiveness of NAMI-A on the metastatic melanoma and suggests that metastasis inhibition is due to the negative modulation of tumor cell invasion processes, a mechanism in which the reduction of the gelatinolitic activity of tumor cells plays a crucial role. PMID- 16368901 TI - Peroxisome proliferator-activated receptor gamma down-regulates receptor for advanced glycation end products and inhibits smooth muscle cell proliferation in a diabetic and nondiabetic rat carotid artery injury model. AB - Diabetes is associated with an increase in circulating advanced glycosylation end products (AGEs) and the increased expression of the receptor for AGEs (RAGE). Inhibition of AGE/RAGE binding through the administration of soluble RAGE (sRAGE) has been shown to decrease neointimal hyperplasia. Peroxisome proliferator activated receptor gamma (PPARgamma), which inhibits neointimal hyperplasia, has been shown to decrease RAGE expression in cultured endothelial cells. We hypothesized that PPARgamma agonists inhibit neointimal hyperplasia via down regulation of RAGE in vivo. Pretreatment of rat aortic smooth muscle cells (SMCs) with PPARgamma agonist rosiglitazone significantly down-regulated RAGE expression and inhibited SMC proliferation in response to the RAGE agonist S100/calgranulins. In vivo studies showed that rosiglitazone decreased RAGE expression and SMC proliferation at 7 days following carotid arterial injury in both diabetic and nondiabetic rats. At 21 days following injury, neointimal formation was significantly decreased in both diabetic and nondiabetic animals that received rosiglitazone. To determine whether inhibition of neointimal formation by PPARgamma activation could fully be accounted for by its down regulation of RAGE, we compared the results obtained in animals treated with sRAGE, PPARgamma activator, and sRAGE + PPARgamma activator. Consistent with PPARgamma working through its effects on RAGE, we found that the addition of PPARgamma activator to sRAGE did not result in any further decrease in neointimal formation. These data demonstrate for the first time that PPARgamma agonists inhibit RAGE expression at sites of arterial injury and suggest that down regulation of RAGE by the PPARgamma activation inhibits neointimal formation in response to arterial injury. PMID- 16368903 TI - Nonpharmacologic and complementary approaches to cancer pain management. AB - Nonpharmacologic interventions are important adjuncts to treatment modalities for patients with cancer pain. A variety of modalities can be used to reduce pain and concomitant mood disturbance and increase quality of life. Physicians may feel relatively uninformed about which modalities have been used for patients with cancer and which have scientific support. This article reviews a few of the nonpharmacologic and complementary and alternative modalities commonly used by patients with cancer pain. It focuses on those modalities that have empirical support or promising preliminary evidence, with the goal of familiarizing physicians with treatment modalities that may complement regular oncologic care. PMID- 16368902 TI - Second-generation inhibitors demonstrate the involvement of p38 mitogen-activated protein kinase in post-transcriptional modulation of inflammatory mediator production in human and rodent airways. AB - The exact role of p38 mitogen-activated protein kinase (MAPK) in the expression of inflammatory cytokines is not clear; it may regulate transcriptionally, post transcriptionally, translationally, or post-translationally. The involvement of one or more of these mechanisms has been suggested to depend on the particular cytokine, the cell type studied, and the specific stimulus used. Interpretation of some of the published data is further complicated by the use of inhibitors such as 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB 203580) used at single, high concentrations. The aim of this study was to determine the impact of two second-generation p38 MAPK inhibitors on the expression of a range of inflammatory cytokines at the gene and protein levels in human cultured cells. Similar assessment of the impact of these compounds on inflammatory cytokine expression in a preclinical in vivo model of airway inflammation was performed. The results in THP-1 cells and primary airway macrophages clearly show that protein expression is inhibited at much lower concentrations of inhibitor than are needed to impact on gene expression. In the rodent model, both compounds, at doses that cause maximal inhibition of cellular recruitment, inhibit tumor necrosis factor alpha (TNFalpha) protein production without impacting on nuclear factor kappaB pathway activation or TNFalpha gene expression. In summary, the data shown here demonstrate that, although at high compound concentrations there is some level of transcriptional regulation, the predominant role of p38 MAPK in cytokine production is at the translational level. These data question whether the effect of p38 inhibitors on gene transcription is related to their potential therapeutic role as anti-inflammatory compounds. PMID- 16368904 TI - Pharmacologic management of cancer pain. AB - Cancer remains the second most common cause of death in the United States despite advances in prevention, early detection, and newer treatment protocols. Pain continues to be the most feared complication of this diagnosis. Numerous studies have shown that when the World Health Organization treatment guidelines are followed, 90% of patients are pain-free. Although evidence is convincing that opioids are effective in the treatment of patients with cancer pain, reluctance to use them continues. Barriers to opioid use are multifactorial, but with education of healthcare providers and patients, treatment to manage pain will be more effective. PMID- 16368905 TI - American Osteopathic Association adopts policies on treatment of patients in pain: an overview. PMID- 16368906 TI - American Osteopathic Association's policy statement on end-of-life care. PMID- 16368907 TI - Becoming an advocate for cancer pain management. AB - Management of cancer pain is still a significant problem in healthcare today despite the fact that cancer pain can be controlled in approximately 90% of patients. Emotional, psychosocial, and spiritual suffering associated with the disease complicates the problem. Guidelines issued by the Agency for Healthcare Research and Quality address management of cancer pain. Pain intensity scales, complementary and alternative methods, and the role of the interdisciplinary care team, as well as the need to provide spiritual support to the patient and family, are included in the discussion. PMID- 16368908 TI - Palliative care: optimizing quality of life. AB - Many developments have occurred in the prevention and treatment of cancer, but death from this disease is still common. According to the World Health Organization, 7 million deaths were due to cancer worldwide in 1999. For dying patients, it is most important to improve quality of life and relieve suffering. Palliative care is defined as the active total care of patients whose disease is not responsive to curative treatment. It encompasses all treatment modalities that are aimed at enhancing quality of life rather than curing disease. Each patient's definition of quality of life is unique. As such, it is important to treat each person as an individual and to continue to view the patient holistically. Controlling cancer-related symptoms can ameliorate the patient's limited remaining time with family and friends. Palliative chemotherapy, rehabilitation, radiation therapy, surgery, and interventional pain management can help to achieve this objective. PMID- 16368909 TI - Report of case: Relapse of condyloma acuminatum and mistrust of physicians in homeless patient. PMID- 16368910 TI - Doctors' dilemma: prescription pain medications. PMID- 16368911 TI - Dynamic duo: Maine-Dartmouth Family Practice Residency program and University of New England College of Osteopathic Medicine. PMID- 16368912 TI - Osteopathic manipulative treatment out of a horse and buggy. PMID- 16368913 TI - The coming influenza pandemic: lessons from the past for the future. PMID- 16368914 TI - Community-based osteopathic manipulative medicine student clinic: changes in curriculum and student confidence levels. AB - The West Virginia School of Osteopathic Medicine in Lewisburg instituted a free, community-based medical clinic in spring 1995 as part of an institution-wide Osteopathic Principles and Practices Integration Project. Under this initiative, osteopathic medical students are provided with 26 hours of supervised experience in osteopathic palpatory diagnosis and osteopathic manipulative treatment. The educational goal of the clinic is to increase the confidence of osteopathic medical students in their technical abilities in using osteopathic manipulative medicine to diagnose and treat community patients. Based on a variety of results from student-completed surveys, the authors conclude that the osteopathic manipulative medicine student clinic at West Virginia School of Osteopathic Medicine has met its goal of increasing student confidence in the application of these skills. PMID- 16368915 TI - Impact of concierge care on healthcare and clinical practice. PMID- 16368916 TI - Report of case: Partial ureteral obstruction masked by diuretics during intraoperative cystoscopy. AB - Injury to the lower urinary tract is a potential complication in all major vaginal and urogynecologic surgical procedures. Several authors have recommended the routine use of intraoperative cystoscopy during urogynecologic procedures. To evaluate possible injury to the lower urinary tract during intraoperative cystoscopy, the concomitant use of diuretics with indigo carmine dye has been advocated; efflux of dye is hypothesized to indicate functional patency of the urinary tract. This report describes a case in which a partial ureteral obstruction was present at the time of intraoperative cystoscopy--despite the observation of diuresis caused by furosemide. This case indicates that the efflux of indigo carmine-stained urine from both ureteral orifices is not conclusive evidence of the absence of ureteral insult during intraoperative cystoscopy. PMID- 16368917 TI - Neurotoxicity related to valganciclovir in a child with impaired renal function: usefulness of therapeutic drug monitoring. AB - OBJECTIVE: To report a case of neurotoxicity related to antiviral drugs, discuss the involvement of concomitant medications, and document the pharmacokinetics of ganciclovir (administered as valganciclovir) in a child with impaired renal function. CASE SUMMARY: A 13-year-old boy with acute lymphoblastic leukemia was treated for cytomegalovirus retinitis with valganciclovir 450 mg every 2 days in the course of hematopoietic stem cell transplantation. Concomitant medication included omeprazole, furosemide, and acetaminophen. During treatment, when creatinine clearance decreased to 20 mL/min, the child presented with acute neurotoxicity, consisting of mental confusion and hallucinations, which resolved when all medications were stopped. Valganciclovir therapeutic monitoring showed high ganciclovir concentrations in the plasma (3.85 microg/mL) and cerebrospinal fluid (2.6 microg/mL) 48 hours after the last valganciclovir dose. After recovery of neurologic function, valganciclovir was resumed at a lower dosage (225 mg twice a week) with therapeutic drug monitoring and was well tolerated. However, the cytomegalovirus infection was not resolved. The leukemia relapsed, and the patient had terminal renal failure and died. The Naranjo probability scale indicated a probable relationship between valganciclovir and neurotoxicity. DISCUSSION: Drugs taken by this child (acyclovir, valganciclovir, omeprazole) have been reported to induce neurotoxicity, with the pharmacokinetics of the first 2 being altered by renal failure. At the time when acyclovir was first administered, symptoms of neurotoxicity were already apparent. Moreover, plasma concentrations of ganciclovir were very high during the course of the neurotoxicity. Thus, the adverse effects seemed related to an overdosage of valganciclovir and were worsened by the addition of acyclovir. CONCLUSIONS: This case is informative because few clinical and pharmacokinetic data are available concerning the use of valganciclovir in children. A study should be performed to determine the proper pediatric dose of valganciclovir with and without renal impairment to prevent the occurrence of adverse effects. PMID- 16368918 TI - Possible interaction between lopinavir/ritonavir and valproic Acid exacerbates bipolar disorder. AB - OBJECTIVE: To describe a case of exacerbated mania potentially related to an interaction between lopinavir/ritonavir and valproic acid (VPA) and propose a mechanism of action for this interaction. CASE SUMMARY: A 30-year-old man with bipolar disorder and HIV initiated treatment with lopinavir/ritonavir, zidovudine, and lamivudine. Prior to beginning therapy with these antiretrovirals, he was receiving VPA 250 mg 3 times daily, with his most recent VPA concentration measured at 495 micromol/L. Twenty-one days after starting antiretroviral treatment, he became increasingly manic. His VPA concentration at admission was 238 micromol/L, a 48% decrease. The daily VPA dose was increased to 1500 mg, and olanzapine was introduced. The VPA concentration following this dose escalation was 392 micromol/L, and the patient improved clinically. DISCUSSION: Fifty percent of VPA is metabolized by glucuronidation, 40% undergoes mitochondrial beta-oxidation, and less than 10% is eliminated by the cytochrome P450 isoenzymes. Ritonavir can induce glucuronidation of several medications including ethinyl estradiol, levothyroxine, and lamotrigine. We believe that ritonavir-mediated induction of VPA glucuronidation resulted in a decrease in VPA concentrations and efficacy. An objective causality assessment suggested that the increased mania was probably related to the decrease in VPA concentration and that a possible interaction exists between lopinavir/ritonavir and VPA. CONCLUSIONS: A potential interaction exists between VPA and all ritonavir-boosted antiretroviral regimens. Clinicians should monitor patients closely for a decreased VPA effect when these medications are given concomitantly. PMID- 16368919 TI - Cyclosporine for severe ulcerative colitis. AB - OBJECTIVE: To evaluate evidence for the use of cyclosporine in treating patients with severe ulcerative colitis. DATA SOURCES: A literature search was performed using MEDLINE, EMBASE, Cochrane Database, and ISI Web of Knowledge (1966-November 2005) with the search terms cyclosporine, cyclosporin A, CsA, ulcerative colitis, UC, inflammatory bowel disease, IBD, steroid-refractory, and immunosuppression. Additional papers were located by hand-searching relevant references. Only human studies in adults and literature published in English were included. DATA SYNTHESIS: Intravenous cyclosporine has been evaluated for the treatment of severe ulcerative colitis in 4 randomized, controlled trials, as well as in many open-label and retrospective studies. Studies that evaluated cyclosporine for severe ulcerative colitis were reviewed. All 4 controlled trials showed an initial positive clinical response as defined by the Crohn's Activity Index when intravenous cyclosporine 4 mg/kg/day was administered as monotherapy or combined with intravenous corticosteroids. One of the 4 trials indicated that high-dose cyclosporine (4 mg/kg/day) has no additional clinical benefit over the low-dose (2 mg/kg/day) and that the lower dose may improve safety related to dose dependent adverse effects. CONCLUSIONS: There is evidence to support the use of intravenous cyclosporine for patients with severe ulcerative colitis who are refractory to corticosteroid therapy. Because most of the adverse effects associated with cyclosporine are dose dependent, therapy should be initiated with the lower 2 mg/kg/day dose. Subsequent doses should be adjusted based on cyclosporine blood concentrations of 150-250 ng/mL. Cyclosporine should be used only to induce remission and serve as a "bridge" to azathioprine or 6 mercaptopurine maintenance therapy. At this time, there are insufficient data to support the long-term use of cyclosporine monotherapy for avoidance of surgery or maintenance of remission. PMID- 16368920 TI - Evolution of drug-drug interactions: a personal viewpoint. PMID- 16368921 TI - Microbiological tests on oral pediatric medicines--requirements for an improved pharmacopoeia monograph. PMID- 16368923 TI - Can olanzapine cause delirium in the elderly? AB - OBJECTIVE: To report a case of delirium probably caused by the atypical antipsychotic olanzapine in a 74-year-old man with dementia. CASE SUMMARY: A 74 year-old white man with a diagnosis of severe dementia of mixed etiology with behavioral disturbances was admitted to an urban teaching hospital for increasing agitation in the context of worsening dementia. Olanzapine 2.5 mg each evening was started for agitation, and the dose was titrated to 5 mg each evening with additional emergent doses. Memantine, an N-methyl-D-aspartate antagonist, was increased from the admission dose of 10 mg/day to 15 mg/day. The patient developed symptoms of delirium on hospital day 4. Neuroleptic malignant syndrome and other causes of delirium were ruled out. Discontinuation of olanzapine resulted in resolution of the delirium. DISCUSSION: Antipsychotic medications are commonly used to treat symptoms of delirium. Atypical antipsychotics are better tolerated in the elderly because of their fewer adverse reactions compared with other antipsychotics. Olanzapine has been successfully used in the treatment of delirium. However, there have been case reports of delirium associated with olanzapine, probably related to its intrinsic anticholinergic effect. Application of the Naranjo probability scale indicated a probable relationship between the onset of delirium and the use of olanzapine in this patient. As of December 1, 2005, this was the second such report of a case in the elderly. CONCLUSIONS: Although olanzapine is useful in the treatment of delirium, elderly patients treated with this drug can develop delirium and hence should be closely monitored. PMID- 16368922 TI - Effects of steady-state lasofoxifene on CYP2D6- and CYP2E1-mediated metabolism. AB - BACKGROUND: Lasofoxifene, a selective estrogen receptor modulator, may be coadministered with other drugs, raising the issue of drug-drug interactions. OBJECTIVE: Using a 7-day, open-label, sequential study to determine whether lasofoxifene at steady-state concentration affects cytochrome P450-mediated drug metabolism. METHODS: Lasofoxifene was tested in 18 postmenopausal women with probe drugs for CYP2E1 and CYP2D6. Changes in CYP2E1 metabolism were measured by the formation clearance of 6-hydroxychlorzoxazone (6-OHCLZ; Cl(f,6-OHCLZ)) following a 250 mg dose of chlorzoxazone in the absence (day 1) and presence (day 6) of lasofoxifene. Changes in the dextromethorphan/dextrorphan urine metabolic ratio (MRDX) measured the effect on CYP2D6 metabolism following a 30 mg dose of dextromethorphan in the absence and presence of lasofoxifene (days 2 and 7). RESULTS: Steady-state lasofoxifene did not affect the formation clearance of 6 OHCLZ or the urinary MRDX. For 6-OHCLZ, the lower boundary (87.12%) of the 90% confidence interval for the ratio (day 6/day 1) of Cl(f,6-OHCLZ) was well above the clinically acceptable ratio of 60%. Both the individual and group mean Cl(f,6 OHCLZ) values were comparable in the absence and presence of lasofoxifene. For MRDX, the upper boundary (129.37%) of the 90% confidence interval for the ratio (day 7/day 2) of MRDX was well below the stipulated ratio of 200%. The individual and mean MRDX values were comparable in the absence and presence of lasofoxifene. Lasofoxifene was well tolerated; adverse events were mild and transient. CONCLUSIONS: Lasofoxifene has no effect on CYP2E1- or CYP2D6-mediated drug metabolism and should not affect drugs metabolized by other cytochrome P450 isoenzymes. PMID- 16368924 TI - Effects of a national health education program on the medication knowledge of the public in Taiwan. AB - BACKGROUND: The inappropriate use of medication and inadequate medication knowledge among the general population has long been a concern in Taiwan. One reason for the deficiencies might be the lack of an active role of pharmacists in educating the public. To rectify the situation, in 2002, the Bureau of Pharmaceutical Affairs, Department of Health of Taiwan, began to sponsor a national effort, titled Community Education Program on Medication Use, to involve the expertise of pharmacists in public education. OBJECTIVE: To evaluate the effects of this education program by analyzing the changes in knowledge of drug therapy among the participating public. METHODS: This was a single-group pre- and post-comparison study. Between September 2003 and January 2004, a total of 955 community residents enrolled in the pharmacist-facilitated education program offered at 31 community universities. The medication knowledge of the participants was evaluated before and after the program. Demographic variables that might affect the education outcomes of the program were also examined. RESULTS: Medication knowledge at baseline was positively correlated with education level and negatively correlated with age. Females were more aware of drug-related information than were males. The participants showed a significant improvement in medication knowledge (p < 0.001) at the end of the program. The baseline knowledge score was the most important determinant of the improvement of the posttest score. CONCLUSIONS: A national education program facilitated by pharmacists can improve the medication knowledge of the participants. Pharmacists should be encouraged to play a proactive role in large-scale health education programs. PMID- 16368925 TI - Chronic hepatitis C virus management: 2000-2005 update. AB - OBJECTIVE: To review recent advances that have significantly changed the management of chronic hepatitis C virus (HCV) infection. DATA SOURCES: A MEDLINE search (2000-July 2005) was conducted using key words such as hepatitis C, interferon, pegylated interferon, and therapy. STUDY SELECTION AND DATA EXTRACTION: All articles pertaining to treatment of chronic HCV infection were identified. Studies evaluating HCV treatment in treatment-naive patients were considered for this review. DATA SYNTHESIS: Over the past several years, response to treatment for chronic HCV infection has significantly improved with the use of pegylated interferon and ribavirin therapy. Treatment response is influenced by HCV genotype and viral load, as well as patient-related factors, including adherence. CONCLUSIONS: Treatment of chronic HCV infection has improved, with overall response rates of approximately 55%. Identification and management of common adverse effects is important in maximizing adherence and response to therapy. Studies are needed to further delineate the optimum treatment of chronic HCV infection in specific patient populations. PMID- 16368926 TI - Quality assessment of dyslipidemia in managed care: current best evidence should be used to benchmark quality. AB - Quality measurements in managed care allow purchasers of health care to distinguish between health plans. Existing measures (Health Plan Employer Data and Information Set goals) for treatment of dyslipidemia provide a limited snapshot about quality of care for members within commercial health plans. Newer evidence (ie, the Heart Protection Study) and consensus guidelines (the National Cholesterol Education Program) expand the definition of high-risk populations and emphasize pharmacotherapy in managing dyslipidemia. We believe that newer evidence and standards provide health plans with the best opportunity to accurately assess the quality of dyslipidemia care for their populations. We propose a broad framework that provides health plans with guidance on developing a new quality measure for dyslipidemia that focuses on pharmacotherapy. PMID- 16368927 TI - Serotonin syndrome induced by fluvoxamine and oxycodone. AB - OBJECTIVE: To report a case of severe serotonergic symptoms following the addition of oxycodone to fluvoxamine. CASE SUMMARY: A 70-year-old woman developed severe serotonergic features, including confusion, nausea, fever, clonus, hyperreflexia, hypertonia, shivering, and tachycardia, following the addition of oxycodone 40 mg twice daily to fluvoxamine 200 mg/day, easily fulfilling diagnostic criteria for serotonin syndrome. Discontinuation of the offending drugs resulted in resolution of her symptoms over 48 hours, and no other cause of the syndrome was identified. Use of the Naranjo probability scale indicated a probable relationship between the serotonergic symptoms and the addition of oxycodone to fluvoxamine therapy. DISCUSSION: Serotonin syndrome is a serious adverse reaction usually due to interactions with serotonergic drugs. There have been only 3 previous reports involving oxycodone. Most previous reports of serotonin syndrome involving analgesics have been associated with meperidine, dextromethorphan, and tramadol. Unlike these synthetic opioids, however, oxycodone does not inhibit the reuptake of serotonin. In addition, there are a number of other possible pharmacologic mechanisms for the interaction we observed. CONCLUSIONS: Monitoring for serotonergic adverse events should be done when oxycodone is given to patients receiving serotonin-reuptake inhibitors. PMID- 16368928 TI - Shaping BMP morphogen gradients in the Drosophila embryo and pupal wing. AB - In the early Drosophila embryo, BMP-type ligands act as morphogens to suppress neural induction and to specify the formation of dorsal ectoderm and amnioserosa. Likewise, during pupal wing development, BMPs help to specify vein versus intervein cell fate. Here, we review recent data suggesting that these two processes use a related set of extracellular factors, positive feedback, and BMP heterodimer formation to achieve peak levels of signaling in spatially restricted patterns. Because these signaling pathway components are all conserved, these observations should shed light on how BMP signaling is modulated in vertebrate development. PMID- 16368930 TI - Hormonal regulation of mummy is needed for apical extracellular matrix formation and epithelial morphogenesis in Drosophila. AB - Many epithelia produce apical extracellular matrices (aECM) that are crucial for organ morphogenesis or physiology. Apical ECM formation relies on coordinated synthesis and modification of constituting components, to enable their subcellular targeting and extracellular assembly into functional matrices. The exoskeleton of Drosophila, the cuticle, is a stratified aECM containing ordered chitin polysaccharide lamellae and proteinaceous layers, and is suited for studies of molecular functions needed for aECM assembly. Here, we show that Drosophila mummy (mmy) mutants display defects in epithelial organisation in conjunction with aberrant deposition of the cuticle and an apical matrix needed for tracheal tubulogenesis. We find that mmy encodes the UDP-N-acetylglucosamine pyrophosphorylase, which catalyses the production of UDP-N-acetylglucosamine, an obligate substrate for chitin synthases as well as for protein glycosylation and GPI-anchor formation. Consequently, in mmy mutants GlcNAc-groups including chitin are severely reduced and modification and subcellular localisation of proteins designated for extracellular space is defective. Moreover, mmy expression is selectively upregulated in epithelia at the time they actively deposit aECM, and is altered by the moulting hormone 20-Hydroxyecdysone, suggesting that mmy is part of a developmental genetic programme to promote aECM formation. PMID- 16368929 TI - The Vg1-related protein Gdf3 acts in a Nodal signaling pathway in the pre gastrulation mouse embryo. AB - The formation of the anterior visceral endoderm (AVE) in the pre-gastrulation mouse embryo represents a crucial event in patterning of the anterior-posterior axis. Here, we show that the transforming growth factor beta (Tgfbeta) family member Gdf3 (growth-differentiation factor 3), a close relative of Xenopus Vg1, resembles the Tgfbeta ligand Nodal in both its signaling activity and its role in AVE formation in vivo. Thus, in cell culture, Gdf3 signaling requires the EGF-CFC co-receptor Cripto and can be inhibited by Lefty antagonists. In Xenopus embryos, Gdf3 misexpression results in secondary axis formation, and induces morphogenetic elongation and mesendoderm formation in animal caps. In mouse embryos, Gdf3 is expressed in the inner cell mass and epiblast, and null mutants frequently exhibit abnormal formation or positioning of the AVE. This phenotype correlates with defects in mesoderm and definitive endoderm formation, as well as abnormal Nodal expression levels. Our findings indicate that Gdf3 acts in a Nodal-like signaling pathway in pre-gastrulation development, and provide evidence for the functional conservation of Vg1 activity in mice. PMID- 16368931 TI - Centrosome destined to decay in starfish oocytes. AB - In contrast to the somatic cell cycle, duplication of the centrioles does not occur in the second meiotic cycle. Previous studies have revealed that in starfish each of the two centrosomes in fully-grown immature oocytes consists of two centrioles with different destinies: one survives and retains its reproductive capacity, and the other is lost after completion of meiosis. In this study, we investigated whether this heterogeneity of the meiotic centrioles is already determined before the re-initiation of meiosis. We prepared a small fragment of immature oocyte containing the four centrioles and fused it electrically with a mature egg in order to transfer two sets of the premeiotic centrioles into the mature cytoplasm. Two asters were present in this conjugate, and in each of them only a single centriole was detected by electron microscopy. In the first mitosis of the conjugate artificially activated without sperm, two division poles formed, each of which doubled in each subsequent round of mitosis. These results indicate that only two of the four premeiotic centrioles survived in the mature cytoplasm and that they retained their reproductive capacity, which suggests that the heterogeneity of the maternal centrioles is determined well before re-initiation of meiosis, and that some factor in the mature cytoplasm is responsible for suppressing the reproductive capacity of the centrioles destined to decay. PMID- 16368932 TI - Retinaldehyde dehydrogenase 2 (RALDH2)-mediated retinoic acid synthesis regulates early mouse embryonic forebrain development by controlling FGF and sonic hedgehog signaling. AB - Although retinoic acid (RA) has been implicated as one of the diffusible signals regulating forebrain development, patterning of the forebrain has not been analyzed in detail in knockout mouse mutants deficient in embryonic RA synthesis. We show that the retinaldehyde dehydrogenase 2 (RALDH2) enzyme is responsible for RA synthesis in the mouse craniofacial region and forebrain between the 8- and 15 somite stages. Raldh2-/- knockout embryos exhibit defective morphogenesis of various forebrain derivatives, including the ventral diencephalon, the optic and telencephalic vesicles. These defects are preceded by regionally decreased cell proliferation in the neuroepithelium, correlating with abnormally low D-cyclin gene expression. Increases in cell death also contribute to the morphological deficiencies at later stages. Molecular analyses reveal abnormally low levels of FGF signaling in the craniofacial region, and impaired sonic hedgehog signaling in the ventral diencephalon. Expression levels of several regulators of diencephalic, telencephalic and optic development therefore cannot be maintained. These results unveil crucial roles of RA during early mouse forebrain development, which may involve the regulation of the expansion of neural progenitor cells through a crosstalk with FGF and sonic hedgehog signaling pathways. PMID- 16368933 TI - p53 suppresses the self-renewal of adult neural stem cells. AB - There is increasing evidence that tumors are heterogeneous and that a subset of cells act as cancer stem cells. Several proto-oncogenes and tumor suppressors control key aspects of stem cell function, suggesting that similar mechanisms control normal and cancer stem cell properties. We show here that the prototypical tumor suppressor p53, which plays an important role in brain tumor initiation and growth, is expressed in the neural stem cell lineage in the adult brain. p53 negatively regulates proliferation and survival, and thereby self renewal, of neural stem cells. Analysis of the neural stem cell transcriptome identified the dysregulation of several cell cycle regulators in the absence of p53, most notably a pronounced downregulation of p21 expression. These data implicate p53 as a suppressor of tissue and cancer stem cell self-renewal. PMID- 16368934 TI - TGFbeta-mediated FGF signaling is crucial for regulating cranial neural crest cell proliferation during frontal bone development. AB - The murine frontal bone derives entirely from the cranial neural crest (CNC) and consists of the calvarial (lateral) aspect that covers the frontal lobe of brain and the orbital aspect that forms the roof of bony orbit. TGFbeta and FGF signaling have important regulatory roles in postnatal calvarial development. Our previous study has demonstrated that conditional inactivation of Tgfbr2 in the neural crest results in severe defects in calvarial development, although the cellular and molecular mechanisms by which TGFbeta signaling regulates the fate of CNC cells during frontal bone development remain unknown. Here, we show that TGFbeta IIR is required for proliferation of osteoprogenitor cells in the CNC derived frontal bone anlagen. FGF acts downstream of TGFbeta signaling in regulating CNC cell proliferation, and exogenous FGF2 rescues the cell proliferation defect in the frontal primordium of Tgfbr2 mutant. Furthermore, the CNC-derived frontal primordium requires TGFbeta IIR to undergo terminal differentiation. However, this requirement is restricted to the developing calvarial aspect of the frontal bone, whereas the orbital aspect forms despite the ablation of Tgfbr2 gene, implying a differential requirement for TGFbeta signaling during the development of various regions of the frontal bone. This study demonstrates the biological significance of TGFbeta-mediated FGF signaling cascade in regulating frontal bone development, suggests that TGFbeta functions as a morphogen in regulating the fate of the CNC-derived osteoblast and provides a model for investigating abnormal craniofacial development. PMID- 16368935 TI - Back to the drawing board on immunohistochemistry and predictive factors. PMID- 16368936 TI - New and surprising insights into pathogenesis of multicentric squamous cancers in the female lower genital tract. PMID- 16368937 TI - Targeting mitochondria emerges as therapeutic strategy. PMID- 16368938 TI - Accelerated approval regulations may need overhaul, panel suggests. PMID- 16368939 TI - Consumer groups look to improve adverse event reporting systems. PMID- 16368940 TI - Stat bite: Lung cancer stage at diagnosis in the United States, 1995-2001. PMID- 16368941 TI - FDA considers tightening regulations for direct-to-consumer advertising. PMID- 16368942 TI - Automated quantitative analysis (AQUA) of in situ protein expression, antibody concentration, and prognosis. AB - BACKGROUND: Disparate results in the immunohistochemistry literature regarding the relationship between biomarker expression and patient outcome decrease the credibility of tissue biomarker studies. We investigated whether some of these disparities result from subjective optimization of antibody concentration. METHODS: We used the automated quantitative analysis (AQUA) system and various concentrations of antibodies against HER2 (1 : 500 to 1 : 8000 dilutions), p53 (1 : 50 to 1 : 800 dilutions), and estrogen receptor (ER; 1 : 100 and 1 : 1000 dilutions) to assess expression of HER2 and p53 in a tissue microarray containing specimens from 250 breast cancer patients with long-term survival data available. HER2 expression in the tissue microarray was also assessed by conventional immunohistochemistry. Relative risk (RR) of disease-specific mortality was assessed for every cutpoint with the X-tile program. Cumulative disease-specific survival was assessed by the Kaplan-Meier method. All statistical tests were two sided. RESULTS: For HER2 and p53 and an optimal cutpoint, when a high antibody concentration (i.e., 1 : 500 dilution) was used with the AQUA system, low expression was associated with poorer survival than high expression; however, when a low antibody concentration (i.e., 1 : 8000 dilution) was used, high expression was associated with poorer survival. For example, for a 1 : 8000 dilution of HER2 antibody and high expression defined as the top 15% of HER2 expression, high HER2 expression was associated with increased disease-specific mortality (RR = 1.98, 95% confidence interval [CI] = 1.21 to 3.23; P = .007), compared with low expression. However, for a 1 : 500 dilution of HER2 antibody and high expression defined as the top 85% of HER2 expression, high HER2 expression was associated with decreased disease-specific mortality (RR = 0.47, 95% CI = 0.29 to 0.76; P = .002), compared with low HER2 expression. CONCLUSIONS: Biomarker antibody concentration appears to dramatically affect the apparent relationship between biomarker expression and outcome. PMID- 16368943 TI - Clonal history of papillomavirus-induced dysplasia in the female lower genital tract. AB - BACKGROUND: Dysplastic lesions of the vagina or the vulva often occur in women who have a previous history of cervical dysplasia. Most lesions in the female lower genital tract are induced by infections with high-risk oncogenic human papillomaviruses (HR-HPVs), including HPV16 and HPV18. HR-HPV genomes frequently integrate into host cell chromosomes at random sites. We analyzed viral integration sites in multiple metachronous lesions of the lower genital tract from women previously treated for HR-HPV-positive cervical dysplasia or cancer to determine whether the metachronous lesions emerged from a single common preexisting dysplastic cell clone or as consequence of independent HR-HPV infection events in the female lower genital tract. METHODS: From among 1500 patients with anogenital lesions, seven patients with high-grade vaginal or vulvar lesions and with a previous history of cervical disease (five with prior high-grade cervical dysplasia and two with a history of cervical cancer) were included in this study. Integration sites of HPV16 or HPV18 in vaginal or vulvar lesions were mapped by an adaptor ligation polymerase chain reaction (PCR) method. The sequence information was used to design an integrate-specific PCR assay that was applied to DNA extracted from archival paraffin-embedded material derived from biopsy samples of cervical lesions. RESULTS: Identical HPV DNA integration loci were found in vaginal or vulvar and cervical samples of all lesions available for four of the five patients with a prior history of high grade cervical dysplasia and for both patients with a history of cervical cancer. CONCLUSIONS: These data indicate that high-grade dysplastic lesions in the female lower genital tract may emerge primarily as monoclonal lesions from a transformed cell population derived from the uterine cervix. PMID- 16368944 TI - In vitro sensitivity to ultraviolet B light and skin cancer risk: a case-control analysis. AB - BACKGROUND: Mutagen sensitivity, measured as mutagen-induced chromatid breaks per cell in primary lymphocytes in vitro, has been used to study susceptibility to various epithelial cancers. Patients with xeroderma pigmentosum are highly sensitive to ultraviolet (UV) light due to inherited defects in DNA repair and have a 1000-fold higher risk of UV-induced skin cancer than the general population. However, an association between UV-induced chromosomal aberrations and risk of skin cancer in the general population has not been established. METHODS: We assessed in vitro UVB-induced chromatid breaks in a hospital-based case-control study. The study included 469 patients with skin cancer (231 with nonmelanoma skin cancer [NMSC] and 238 with cutaneous malignant melanoma [CMM]) and 329 cancer-free control subjects. Multivariable logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: Compared with the frequency of UVB induced chromatid breaks per cell in control subjects (mean = 0.28 breaks per cell, 95% CI = 0.27 to 0.30), that in NMSC patients (basal cell carcinoma [BCC], n = 143, mean = 0.36 breaks per cell, 95% CI = 0.33 to 0.39 and squamous cell carcinoma [SCC], n = 88, mean = 0.35 breaks per cell, 95% CI = 0.32 to 0.38) was higher (P = .001 and P < .001, respectively), but that in CMM case patients (mean = 0.30 breaks per cell, 95% CI = 0.28 to 0.33) was not (P = .22). A frequency of chromatid breaks per cell above the median of control subjects was associated with nearly threefold increased risks for BCC (OR = 2.78, 95% CI = 1.79 to 4.30) and SCC (OR = 2.62, 95% CI = 1.50 to 4.60), but not with an increased risk of CMM. A dose-response relationship was evident between mutagen sensitivity and risk for both BCC (Ptrend < .001) and SCC (Ptrend < .001). Multiplicative interactions between mutagen sensitivity and sun exposure variables on risk, particularly for sunburn in BCC and hair color, tanning ability, and family history of skin cancer in SCC, were seen for NMSC but not CMM. CONCLUSIONS: UVB induced mutagen sensitivity may play a role in susceptibility to NMSC but not to CMM. PMID- 16368945 TI - Baseline chest radiograph for lung cancer detection in the randomized Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. AB - BACKGROUND: The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial was initiated in 1992 to examine cause-specific mortality reduction from screening for these four cancers in men and women. We report lung cancer detection results of the baseline screening round. METHODS: Of the 154,942 participants enrolled, who were aged 55-74 years with no history of PLCO cancers, 77,465 were randomly assigned to the intervention arm. Current or former smokers and never smokers in this arm received an initial single-view posterior-anterior chest radiograph. RESULTS: In the initial screen, 5991 (8.9%, 95% confidence interval [CI] = 8.7% to 9.2%) of radiographs were suspicious for lung cancer: 8.2% (95% CI = 7.9% to 8.5%) for women and 9.6% (95% CI = 9.3% to 10.0%) for men. Rates were highest for older age groups and for smokers. Among those 5991 participants with a positive screen, 206 (3.4%, 95% CI = 3.0% to 3.9%) underwent biopsy examination, 126 (61.2%, 95% CI = 54.5% to 67.8%) of whom were diagnosed with lung cancer within 12 months of the screen (59 in women and 67 in men). The positive predictive value was 2.1% (95% CI = 1.7% to 2.5%), and 1.9 lung cancers were detected per 1000 screens. Among these cancers, 44% (95% CI = 35% to 52%) were stage I non-small-cell lung cancer. High rates of lung cancer were found in current smokers (6.3 per 1000 screens) and in former smokers who had smoked within the past 15 years (4.9 per 1000 screens). The lung cancer detection rate among never smokers was 0.4 per 1000 screens; this group accounted for 11% (95% CI = 5.6% to 16.6%) of the cancers identified. CONCLUSIONS: In the baseline screen, nearly half the cancers were stage I. Whether this experience results in a reduction in lung cancer mortality is yet to be seen. PMID- 16368946 TI - Tumor-cell homing to lymph nodes and bone marrow and CXCR4 expression in esophageal cancer. AB - BACKGROUND: The chemokine and bone marrow-homing receptor CXCR4 has been implicated in metastatic dissemination of various cancers. We investigated CXCR4 expression in esophageal cancer specimens and its association with survival, lymph node microinvolvement, and bone marrow micrometastasis. METHODS: We analyzed frozen tumor specimens from 136 patients with completely resected esophageal cancer for CXCR4 expression by immunohistochemistry. Lymph node microinvolvement and bone marrow micrometastasis were assessed by immunohistochemistry with monoclonal antibodies Ber-EP4 (against epithelial cell adhesion molecule) and pancytokeratin A45-B/B3 (against several cytokeratins), respectively. Associations between CXCR4 expression and clinicopathologic features, including tumor stage, histologic grade, lymph node metastasis and microinvolvement, bone marrow micrometastasis, and survival, were investigated with Fisher's test, log-rank test, and Cox multivariable analysis. All statistical tests were two-sided. RESULTS: CXCR4 protein was expressed in 75 (55%) of 136 esophageal tumors examined. CXCR4 expression was statistically significantly associated with reduced median overall and disease-specific survival, compared with CXCR4 nonexpression (P < .001; log-rank test). The median overall survival of patients with CXCR4-positive tumors was 20 months and with CXCR4-negative tumors, 76 months (difference = 56 months, 95% confidence interval [CI] = 4 to 108 months; P < .001). The median disease-specific survival of patients with CXCR4-positive tumors was 25 months and with CXCR4-negative tumors was 97 months (difference = 72 months, 95% CI = 34 to 110 months; P < .001). CXCR4 expression was statistically significantly associated with increased lymph node microinvolvement (P < .001) and with increased bone marrow micrometastasis (P < .001). In multivariable analysis, CXCR4 expression, compared with its nonexpression, was identified as the independent variable that was most strongly associated with reduced disease-specific survival (relative risk [RR] of death = 2.03, 95% CI = 1.20 to 3.41; P = .008) and overall survival (RR of death = 2.18, 95% CI = 1.33 to 3.59; P = .002). CONCLUSION: CXCR4 expression was associated with poor clinical outcome in esophageal cancer patients. CXCR4 may have a role in early metastatic spread because its expression was associated with micrometastases to both the lymph nodes and bone marrow. Thus, CXCR4 should be explored further as a target for adjuvant therapy for micrometastatic disease. PMID- 16368947 TI - Effects of prolonged storage of whole plasma or isolated plasma DNA on the results of circulating DNA quantification assays. AB - Analysis of molecular markers in biological fluids has been proposed as a tool for early detection and monitoring of cancer. Circulating plasma DNA concentrations have been found to be higher in cancer patients than in cancer free control subjects, but little is known about the effect of specimen storage on plasma DNA concentrations. Here we investigated the impact of long-term storage of both plasma samples and purified plasma DNA on the reproducibility of plasma DNA quantification as determined using real-time polymerase chain reaction analysis. The analysis was performed on samples from a subset of 34 lung cancer patients and 28 matched control subjects selected from 200 subjects in our previously published case-control study and from 117 cancer-free smokers enrolled in a lung cancer screening program. Two samples of plasma and isolated DNA were assessed for each patient, with a median of 41 months between the first and second assessments for participants in the case-control study and 9 months for participants in the screening study. DNA levels declined substantially between the two assessments at an average rate of approximately 30% per year. These data provide valuable information for the rational planning of retrospective studies of banked series of biological samples, particularly if collected over a long period of time, as can occur in large clinical trials. PMID- 16368948 TI - Re: Limits of predictive models using microarray data for breast cancer clinical treatment outcome. PMID- 16368949 TI - Re: Role of detection method in predicting breast cancer survival: analysis of randomized screening trials. PMID- 16368950 TI - Re: Reporting recommendations for tumor marker prognostic studies (REMARK). PMID- 16368951 TI - Functional expression of the CD163 scavenger receptor on acute myeloid leukemia cells of monocytic lineage. AB - The hemoglobin-haptoglobin (Hb-Hp) scavenger receptor CD163 is a monocyte/macrophage-restricted surface antigen, whose expression is strongly up regulated by glucocorticoids. We have previously shown that CD163 is expressed by acute myeloid leukemia (AML) cells of monocytic lineage. Herein, we expand this finding by demonstrating constitutive and glucocorticoid-enhanced CD163 expression on French-American-British M4/M5 AML cells, and leukemic blasts of other AML subtypes and normal hematopoietic progenitor cells do not express CD163. We provide evidence that the functional characteristics of CD163 are preserved on malignant cells by showing the capability of types M4/M5 blast cells to internalize Hb-Hp by a CD163-mediated mechanism. Together, our results identify CD163 as a potential target for therapeutic intervention. It is important that CD163 does not appear to be released from leukemic blasts under noninflammatory conditions, thus reducing the probability of off-target side effects as a result of competitive binding of potential therapeutic ligands to nonmembrane-bound CD163. PMID- 16368953 TI - Capsule-mediated immune evasion: a new hypothesis explaining aspects of typhoid fever pathogenesis. PMID- 16368955 TI - Role of hyaluronidase in subcutaneous spread and growth of group A streptococcus. AB - Group A streptococcus (GAS) depends on a hyaluronic acid (HA) capsule to evade phagocytosis and to interact with epithelial cells. Paradoxically, GAS also produces hyaluronidase (Hyl), an enzyme that cleaves HA. A common assumption is that Hyl digests structurally identical HA in human tissue to promote bacterial spread. We inactivated the gene encoding extracellular hyaluronidase, hylA, in a clinical Hyl(+) isolate. Hyl(+) and an isogenic Hyl(-) mutant were injected subcutaneously into mice with or without high-molecular-weight dextran blue. The Hyl(-) strain produced small lesions with dye concentrated in close proximity. The Hyl(+) strain produced identical lesions, but the dye diffused subcutaneously. However, Hyl(+) bacteria were not isolated from unaffected skin stained by dye diffusion. Thus, Hyl digests tissue HA and facilitates spread of large molecules but is not sufficient to cause subcutaneous diffusion of bacteria or to affect lesion size. GAS capsule expression was assayed periodically during broth culture and was reduced in Hyl(+) strains relative to Hyl(-) strains at the onset and the end of active capsule synthesis but not during peak synthesis in mid-exponential phase. Thus, Hyl is not sufficiently active to remove capsule during peak synthesis. To demonstrate a possible nutritional role for Hyl, GAS was shown to grow with N-acetylglucosamine but not d-glucuronic acid (both components of HA) as a sole carbon source. However, only Hyl(+) strains could grow utilizing HA as a sole carbon source, suggesting that Hyl may permit the organism to utilize host HA or its own capsule as an energy source. PMID- 16368954 TI - Role of sphingolipids in microbial pathogenesis. PMID- 16368956 TI - The cell-specific phenotype of the polymorphic vacA midregion is independent of the appearance of the cell surface receptor protein tyrosine phosphatase beta. AB - There are two alleles, m1 and m2, of the midregion of the vacuolating cytotoxin gene (vacA) of Helicobacter pylori which code for toxins with different cell specificities. Here we describe the construction of five chimeric strains in which regions of vacA were exchanged between the two genotypes. By analyzing the toxicity of these strains for HeLa and RK13 cells we have confirmed that a 148 amino-acid region determines the phenotypic differences between the two forms of the protein and that this entire region is important for cytotoxicity. Furthermore, we have used our chimeric strains to investigate whether variations in the midregion of VacA have an effect on phorbol 12-myristate 13-acetate (PMA) induced VacA sensitivity in HL-60 cells. The PMA-induced VacA sensitivity of HL 60 cells has been previously associated with the appearance of the cell surface receptor protein tyrosine phosphatase beta (RPTPbeta). Our data indicate that both the m1 and m2 forms of VacA are able to utilize RPTPbeta, and the cell specific phenotype of the midregion is independent of the presence of RPTPbeta. It appears that another as-yet-unidentified receptor exists in HL-60 cells that accounts for the m2 phenotype in this cell line. Also, by studying the effect of PMA on levels of RPTPbeta in other cell lines and toxicity of VacA in these cell lines we have shown that RPTPbeta does not play a major role in the vacuolation of HeLa cells. PMID- 16368957 TI - Dihydrolipoamide acyltransferase is critical for Mycobacterium tuberculosis pathogenesis. AB - Mycobacterium tuberculosis has evolved to persist in host macrophages, where it faces a nutrient-poor environment and is exposed to oxidative and nitrosative stress. To defend itself against oxidative/nitrosative stress, M. tuberculosis expresses an NADH-dependent peroxidase and peroxynitrite reductase that is encoded by ahpC, ahpD, lpd, and dlaT. In addition to its central role in the peroxynitrite reductase complex, dlaT (Rv2215) also encodes the E2 component of pyruvate dehydrogenase. Here we demonstrate that inactivation of dlaT in the chromosome of H37Rv resulted in a mutant (H37RvDeltadlaT) that displayed phenotypes associated with DlaT's role in metabolism and in defense against nitrosative stress. The H37RvDeltadlaT strain showed retarded growth in vitro and was highly susceptible to killing by acidified sodium nitrite. Mouse macrophages readily killed intracellular H37RvDeltadlaT organisms, and in mice dlaT was required for full virulence. PMID- 16368958 TI - The global regulator Ler is necessary for enteropathogenic Escherichia coli colonization of Caenorhabditis elegans. AB - Enteropathogenic Escherichia coli (EPEC) is an important cause of infant diarrhea in developing countries and is useful for general investigations of the bacterial infection process. However, the study of the molecular pathogenesis of EPEC has been hampered by the lack of genetically tractable, convenient animal models. We have therefore developed the use of the nematode Caenorhabditis elegans as a small animal model of infection for this diarrheal pathogen. We found that nematodes died faster on nematode growth medium in the presence of EPEC pathogens than in the presence of the laboratory control strain MG1655. Increased numbers of pathogens in the gut, determined by standard plate count assays and fluorescence microscopy using green fluorescent protein-expressing bacteria, correlated with killing. Deletion of the gene encoding the global regulator Ler severely reduced the ability of EPEC to colonize the nematode gut and could be complemented by providing the ler gene on a multicopy plasmid in trans. Neither the type III secretion system nor the type IV bundle-forming pilus was required for colonization. Combined, the similarities and distinct differences between EPEC infection of nematodes and that of humans offer a unique opportunity to study several stages of the infection process, namely, attachment, colonization, and persistence, in a genetically tractable, inexpensive, and convenient in vivo system. PMID- 16368959 TI - Inhibition of chlamydiae by primary alcohols correlates with the strain-specific complement of plasticity zone phospholipase D genes. AB - Members of the genus Chlamydia are obligate intracellular pathogens that have a unique biphasic developmental cycle and interactions with host cells. Many genes that dictate host infection tropism and, putatively, pathogenic manifestations of disease are clustered in a hypervariable region of the genome termed the plasticity zone (PZ). Comparative genomics studies have determined that an uncharacterized family of PZ genes encoding orthologs of eukaryotic and prokaryotic members of the phospholipase D (PLD) enzyme family varies among chlamydiae. Here, we show that the PZ PLD (pzPLD) of Chlamydia trachomatis are transcribed during both normal and persistent infection and that the corresponding PLD proteins are predominantly localized in reticulate bodies on the inner leaflet of the inclusion membrane. Further, we show that strains of chlamydiae encoding the pzPLD, but not a strain lacking these genes, are inhibited by primary alcohols, potent PLD inhibitors, during growth in HeLa 229 cells. This inhibitory effect is amplified approximately 5,000-fold during recovery from persistent infection. These findings suggest that the chlamydial pzPLD may be important, strain-specific, pathogenesis factors in vivo. PMID- 16368960 TI - Caspase and bid involvement in Clostridium difficile toxin A-induced apoptosis and modulation of toxin A effects by glutamine and alanyl-glutamine in vivo and in vitro. AB - Clostridium difficile is the leading cause of nosocomial bacterial diarrhea. Glutamine and its stable and highly soluble derivative alanyl-glutamine, have been beneficial in models of intestinal injury. In this study, we extend our work on the mechanisms of Clostridium difficile toxin A (TxA)-induced apoptosis in human intestinal epithelial T84 cells and evaluate the effects of glutamine and alanyl-glutamine on TxA-induced apoptosis in vitro and disruption of ileal mucosa in vivo. T84 cells were incubated with TxA (100 ng/ml) in medium with or without glutamine or alanyl-glutamine (3 to 100 mM). Apoptosis was evaluated by DNA fragmentation in vitro and the terminal deoxynucleotidyltransferase-mediated dUTP biotin nick end-labeling method in vivo. Caspase and Bid involvement were investigated by Western blotting. Ligated rabbit ileal loops were used for the evaluation of intestinal secretion, mucosal disruption, and apoptosis. TxA induced caspases 6, 8, and 9 prior to caspase 3 activation in T84 cells and induced Bid cleavage by a caspase-independent mechanism. Glutamine or alanyl glutamine significantly reduced TxA-induced apoptosis of T84 cells by 47% and inhibited activation of caspase 8. Both glutamine and alanyl-glutamine reduced TxA-induced ileal mucosal disruption and secretion. Altogether, we further delineated the apoptosis-signaling cascade induced by TxA in T84 cells and demonstrated the protective effects of glutamine and alanyl-glutamine. Glutamine and alanyl-glutamine inhibited the apoptosis of T84 cells by preventing caspase 8 activation and reduced TxA-induced intestinal secretion and disruption. PMID- 16368961 TI - Dissection of ESAT-6 system 1 of Mycobacterium tuberculosis and impact on immunogenicity and virulence. AB - The dedicated secretion system ESX-1 of Mycobacterium tuberculosis encoded by the extended RD1 region (extRD1) assures export of the ESAT-6 protein and its partner, the 10-kDa culture filtrate protein CFP-10, and is missing from the vaccine strains M. bovis BCG and M. microti. Here, we systematically investigated the involvement of each individual ESX-1 gene in the secretion of both antigens, specific immunogenicity, and virulence. ESX-1-complemented BCG and M. microti strains were more efficiently engulfed by bone-marrow-derived macrophages than controls, and this may account for the enhanced in vivo growth of ESX-1-carrying strains. Inactivation of gene pe35 (Rv3872) impaired expression of CFP-10 and ESAT-6, suggesting a role in regulation. Genes Rv3868, Rv3869, Rv3870, Rv3871, and Rv3877 encoding an ATP-dependent chaperone and translocon were essential for secretion of ESAT-6 and CFP-10 in contrast to ppe68 Rv3873 and Rv3876, whose inactivation did not impair secretion of ESAT-6. A strict correlation was found between ESAT-6 export and the generation of ESAT-6 specific T-cell responses in mice. Furthermore, ESAT-6 secretion and specific immunogenicity were almost always correlated with enhanced virulence in the SCID mouse model. Only loss of Rv3865 and part of Rv3866 did not affect ESAT-6 secretion or immunogenicity but led to attenuation. This suggests that Rv3865/66 represent a new virulence factor that is independent from ESAT-6 secretion. The present study has allowed us to identify new aspects of the extRD1 region of M. tuberculosis and to explore its role in the pathogenesis of tuberculosis. PMID- 16368962 TI - Interaction of Cryptosporidium hominis and Cryptosporidium parvum with primary human and bovine intestinal cells. AB - Cryptosporidiosis in humans is caused by the zoonotic pathogen Cryptosporidium parvum and the anthroponotic pathogen Cryptosporidium hominis. To what extent the recently recognized C. hominis species differs from C. parvum is unknown. In this study we compared the mechanisms of C. parvum and C. hominis invasion using a primary cell model of infection. Cultured primary bovine and human epithelial intestinal cells were infected with C. parvum or C. hominis. The effects of the carbohydrate lectin galactose-N-acetylgalactosamine (Gal/GalNAc) and inhibitors of cytoskeletal function and signal transduction mechanisms on entry of the parasites into host cells were tested. HCT-8 cells (human ileocecal adenocarcinoma cells) were used for the purpose of comparison. Pretreatment of parasites with Gal/GalNAc inhibited entry of C. parvum into HCT-8 cells and primary bovine cells but had no effect on entry of either C. parvum or C. hominis into primary human cells or on entry of C. hominis into HCT-8 cells. Both Cryptosporidium species entered primary cells by a protein kinase C (PKC)- and actin-dependent mechanism. Staurosporine, in particular, attenuated infection, likely through a combination of PKC inhibition and induction of apoptosis. Diversity in the mechanisms used by Cryptosporidium species to infect cells of different origins has important implications for understanding the relevance of in vitro studies of Cryptosporidium pathogenesis. PMID- 16368963 TI - The Brucella suis type IV secretion system assembles in the cell envelope of the heterologous host Agrobacterium tumefaciens and increases IncQ plasmid pLS1 recipient competence. AB - Pathogenic Brucella species replicate within mammalian cells, and their type IV secretion system is essential for intracellular survival and replication. The options for biochemical studies on the Brucella secretion system are limited due to the rigidity of the cells and biosafety concerns, which preclude large-scale cell culture and fractionation. To overcome these problems, we heterologously expressed the Brucella suis virB operon in the closely related alpha(2) proteobacterium Agrobacterium tumefaciens and showed that the VirB proteins assembled into a complex. Eight of the twelve VirB proteins were detected in the membranes of the heterologous host with specific antisera. Cross-linking indicated protein-protein interactions similar to those in other type IV secretion systems, and the results of immunofluorescence analysis supported the formation of VirB protein complexes in the cell envelope. Production of a subset of the B. suis VirB proteins (VirB3-VirB12) in A. tumefaciens strongly increased its ability to receive IncQ plasmid pLS1 in conjugation experiments, and production of VirB1 further enhanced the conjugation efficiency. Plasmid recipient competence correlated with periplasmic leakage and the detergent sensitivity of A. tumefaciens, suggesting a weakening of the cell envelope. Heterologous expression thus permits biochemical characterization of B. suis type IV secretion system assembly. PMID- 16368964 TI - Sexual dimorphism in the control of amebic liver abscess in a mouse model of disease. AB - Amebic liver abscess (ALA) is the most common extraintestinal manifestation of human infection by the enteric protozoan parasite Entamoeba histolytica. In contrast to intestinal infection, ALA greatly predominates in males but is rare in females. Since humans are the only relevant host for E. histolytica, experimental studies concerning this sexual dimorphism have been hampered by the lack of a suitable animal model. By serial liver passage of cultured E. histolytica trophozoites in gerbils and mice, we generated amebae which reproducibly induce ALA in C57BL/6 mice. Interestingly, all animals developed ALA, but the time courses of abscess formation differed significantly between the genders. Female mice were able to clear the infection within 3 days, whereas in male mice the parasite could be recovered for at least 14 days. Accordingly, male mice showed a prolonged time of recovery from ALA. Immunohistology of abscesses revealed that polymorphonuclear leukocytes and macrophages were the dominant infiltrates, but in addition, gamma,delta-T cells, NK cells, and natural killer T (NKT) cells were also present at early times during abscess development, whereas conventional alpha,beta-T cells appeared later, when female mice had already cleared the parasite. Interestingly, male and female mice differed in early cytokine production in response to ameba infection. Enzyme-linked immunospot assays performed with spleen cells of infected animals revealed significantly higher numbers of interleukin-4-producing cells in male mice but significantly higher numbers of gamma interferon (IFN-gamma)-producing cells in female mice. Early IFN-gamma production and the presence of functional NKT cells were found to be important for the control of hepatic amebiasis as application of an IFN-gamma neutralizing monoclonal antibody or the use of NKT knockout mice (Valpha14iNKT, Jalpha 18(-/-)) dramatically increased the size of ALA in female mice. In addition, E. histolytica trophozoites could be reisolated from liver abscesses of Jalpha18(-/-) mice on day 7 postinfection, when wild-type mice had already cleared the parasite. These data suggest that the sexual dimorphism in the control of ALA is due to gender-specific differences in early cytokine production mediated at least in part by NKT cells in response to E. histolytica infection of the liver. PMID- 16368965 TI - The chemokines CXCL9 and CXCL10 promote a protective immune response but do not contribute to cardiac inflammation following infection with Trypanosoma cruzi. AB - The expression of chemokines within the heart during experimental infection of susceptible mice with the Colombiana strain of Trypanosoma cruzi was characterized in an attempt to determine a functional role for these molecules in both host defense and disease. Analysis of chemokine transcripts revealed that CXC chemokine ligand 9 (CXCL9) and CXCL10, as well as CC chemokine ligand 2 (CCL2) and CCL5, were prominently expressed during acute disease, whereas transcripts for CXCL9, CXCL10, and CCL5 remained elevated during chronic infection. Inflammatory macrophages present within the heart were the primary cellular source of these chemokines following T. cruzi infection. Peak chemokine expression levels coincided with increased gamma interferon expression and inflammation within the heart, suggesting a role for these molecules in both host defense and disease. Indeed, simultaneous treatment of T. cruzi-infected mice with neutralizing antibodies specific for CXCL9 and CXCL10 resulted in an increased parasite burden that was sustained out to 50 days p.i. Antibody targeting either CXCL10 or CCL5 did not change either T. cruzi burden within the heart nor attenuate the severity of cardiac inflammation at any time point examined, while targeting CXCL9 in combination with CXCL10 resulted in increased parasite burden. Collectively, these studies imply that CXCL9 and CXCL10 signaling enhances immune responses following parasite infection. However, antibody targeting of CXCL9 and CXCL10, or CXCL10 alone, or CCL5 alone does not directly modulate the inflammatory response within the heart, suggesting that other proinflammatory factors are able to regulate inflammation in this tissue in response to T. cruzi infection. PMID- 16368966 TI - The CC chemokine receptor 5 is important in control of parasite replication and acute cardiac inflammation following infection with Trypanosoma cruzi. AB - Infection of susceptible mice with the Colombiana strain of Trypanosoma cruzi results in an orchestrated expression of chemokines and chemokine receptors within the heart that coincides with parasite burden and cellular infiltration. CC chemokine receptor 5 (CCR5) is prominently expressed during both acute and chronic disease, suggesting a role in regulating leukocyte trafficking and accumulation within the heart following T. cruzi infection. To better understand the functional role of CCR5 and its ligands with regard to both host defense and/or disease, CCR5(-/-) mice were infected with T. cruzi, and the disease severity was evaluated. Infected CCR5(-/-) mice develop significantly higher levels of parasitemia (P < or = 0.05) and cardiac parasitism (P < or = 0.01) during acute infection that correlated with reduced survival. Further, we show that CCR5 is essential for directing the migration of macrophages and T cells to the heart early in acute infection with T. cruzi. In addition, data are provided demonstrating that CCR5 does not play an essential role in maintaining inflammation in the heart during chronic infection. Collectively, these studies clearly demonstrate that CCR5 contributes to the control of parasite replication and the development of a protective immune response during acute infection but does not ultimately participate in maintaining a chronic inflammatory response within the heart. PMID- 16368967 TI - Binding and internalization of glucuronoxylomannan, the major capsular polysaccharide of Cryptococcus neoformans, by murine peritoneal macrophages. AB - Glucuronoxylomannan (GXM), the major component of the capsular polysaccharide of Cryptococcus neoformans, is essential to virulence of the yeast. Previous studies found that the interaction between GXM and phagocytic cells has biological consequences that may contribute to the pathogenesis of cryptococcosis. We found that GXM binds to and is taken up by murine peritoneal macrophages. Uptake is dose and time dependent. Examination of the sites of GXM accumulation by immunofluorescence microscopy showed that the pattern was discontinuous and punctate both on the surfaces of macrophages and at intracellular depots. Although resident macrophages showed appreciable accumulation of GXM, uptake was greatest with thioglycolate-elicited macrophages. A modest stimulation of GXM binding followed treatment of resident macrophages with phorbol 12-myristate 13 acetate, but treatment with lipopolysaccharide or gamma interferon alone or in combination had no effect. Accumulation of GXM was critically dependent on cytoskeleton function; a near complete blockade of accumulation followed treatment with inhibitors of actin. GXM accumulation by elicited macrophages was blocked by treatment with inhibitors of tyrosine kinase, protein kinase C, and phospholipase C, but not by inhibitors of phosphatidylinositol 3-kinase, suggesting a critical role for one or more signaling pathways in the macrophage response to GXM. Taken together, the results demonstrate that it is possible to experimentally enhance or suppress binding of GXM to macrophages, raising the possibility for regulation of the interaction between this essential virulence factor and binding sites on cells that are central to host resistance. PMID- 16368968 TI - Trypanosoma cruzi infection induces proliferation of vascular smooth muscle cells. AB - Trypanosoma cruzi infection causes cardiomyopathy and vasculopathy. Previous studies have demonstrated that infection of human umbilical vein endothelial and smooth muscle cells resulted in activation of extracellular signal-regulated kinase (ERK). In the present study, smooth muscle cells were infected with trypomastigotes, and immunoblot analysis revealed an increase in the expression of cyclin D1 and proliferating cell nuclear antigen (PCNA), important mediators of smooth muscle cell proliferation. Interestingly, after infection, the expression of caveolin-1 was reduced in both human umbilical vein endothelial cells and smooth muscle cells. Immunoblot and immunohistochemical analyses of lysates of carotid arteries obtained from infected mice revealed increased expression of PCNA, cyclin D1, its substrate, phospho-Rb (Ser780), and phospho ERK1/2. The expression of the cyclin-dependent kinase inhibitor p21(Cip1/Waf1), caveolin-1, and caveolin-3 was reduced in carotid arteries obtained from infected mice. There was an increase in the abundance of pre-pro-endothelin-1 mRNA in the carotid artery and aorta from infected mice. The ET(A) receptor was also elevated in infected arteries. ERK activates endothelin-1, which in turn exerts positive feedback activating ERK, and cyclin D1 is a downstream target of both endothelin 1 and ERK. There was significant incorporation of bromodeoxyuridine into smooth muscle cell DNA when treatment was with conditioned medium obtained from infected endothelial cells. Taken together, these data suggest that T. cruzi infection stimulates smooth muscle cell proliferation and is likely a result of the upregulation of the ERK-cyclin D1-endothelin-1 pathway. PMID- 16368969 TI - Transcriptional profiling of Mycoplasma hyopneumoniae during heat shock using microarrays. AB - Bacterial pathogens undergo stress during host colonization and disease processes. These stresses result in changes in gene expression to compensate for potentially lethal environments developed in the host during disease. Mycoplasma hyopneumoniae colonizes the swine epithelium and causes a pneumonia that predisposes the host to enhanced disease from other pathogens. How M. hyopneumoniae responds to changing environments in the respiratory tract during disease progression is not known. In fact, little is known concerning the capabilities of mycoplasmas to respond to changing growth environments. With limited genes, mycoplasmas are thought to possess only a few mechanisms for gene regulation. A microarray consisting of 632 of the 698 open reading frames of M. hyopneumoniae was constructed and used to study gene expression differences during a temperature shift from 37 degrees C to 42 degrees C, a temperature swing that might be encountered during disease. To enhance sensitivity, a unique hexamer primer set was employed for generating cDNA from only mRNA species. Our analysis identified 91 genes that had significant transcriptional differences in response to heat shock conditions (P < 0.01) with an estimated false-discovery rate of 4 percent. Thirty-three genes had a change threshold of 1.5-fold or greater. Many of the heat shock proteins previously characterized in other bacteria were identified as significant in this study as well. A proportion of the identified genes (54 of 91) currently have no assigned function. PMID- 16368970 TI - Phage library screening for the rapid identification and in vivo testing of candidate genes for a DNA vaccine against Mycoplasma mycoides subsp. mycoides small colony biotype. AB - A new strategy for rapidly selecting and testing genetic vaccines has been developed, in which a whole genome library is cloned into a bacteriophage lambda ZAP Express vector which contains both prokaryotic (P(lac)) and eukaryotic (P(CMV)) promoters upstream of the insertion site. The phage library is plated on Escherichia coli cells, immunoblotted, and probed with hyperimmune and/or convalescent-phase antiserum to rapidly identify vaccine candidates. These are then plaque purified and grown as liquid lysates, and whole bacteriophage particles are then used directly to immunize the host, following which P(CMV) driven expression of the candidate vaccine gene occurs. In the example given here, a semirandom genome library of the bovine pathogen Mycoplasma mycoides subsp. mycoides small colony (SC) biotype was cloned into lambda ZAP Express, and two strongly immunodominant clones, lambda-A8 and lambda-B1, were identified and subsequently tested for vaccine potential against M. mycoides subsp. mycoides SC biotype-induced mycoplasmemia. Sequencing and immunoblotting indicated that clone lambda-A8 expressed an isopropyl-beta-d-thiogalactopyranoside (IPTG)-inducible M. mycoides subsp. mycoides SC biotype protein with a 28-kDa apparent molecular mass, identified as a previously uncharacterized putative lipoprotein (MSC_0397). Clone lambda-B1 contained several full-length genes from the M. mycoides subsp. mycoides SC biotype pyruvate dehydrogenase region, and two IPTG-independent polypeptides, of 29 kDa and 57 kDa, were identified on immunoblots. Following vaccination, significant anti-M. mycoides subsp. mycoides SC biotype responses were observed in mice vaccinated with clones lambda-A8 and lambda-B1. A significant stimulation index was observed following incubation of splenocytes from mice vaccinated with clone lambda-A8 with whole live M. mycoides subsp. mycoides SC biotype cells, indicating cellular proliferation. After challenge, mice vaccinated with clone lambda-A8 also exhibited a reduced level of mycoplasmemia compared to controls, suggesting that the MSC_0397 lipoprotein has a protective effect in the mouse model when delivered as a bacteriophage DNA vaccine. Bacteriophage-mediated immunoscreening using an appropriate vector system offers a rapid and simple technique for the identification and immediate testing of putative candidate vaccines from a variety of pathogens. PMID- 16368971 TI - Antigen discovery: a postgenomic approach to leprosy diagnosis. AB - Leprosy is an infectious, neurodegenerative disease of humans caused by Mycobacterium leprae. Despite effective control programs, the incidence of leprosy remains stubbornly high, suggesting that transmission may be more common than expected. The rationale of this work was to use bioinformatics and comparative genomics to identify potentially antigenic proteins for diagnostic purposes. This approach defined three classes of proteins: those restricted to M. leprae (class I), those present in M. leprae with orthologues in other organisms besides mycobacteria (class II), and exported or surface-exposed proteins (class III). Twelve genes (two class I, four class II, and six class III proteins) were cloned in Escherichia coli, and their protein products were purified. Six of these proteins were detected in cell extracts of M. leprae by immunoblotting. The immunogenicity of each recombinant protein was then investigated in leprosy patients by measuring the reactivity of circulating antibody and gamma interferon (IFN-gamma) responses in T-cell restimulation assays. Several class II and class III proteins were recognized by circulating antibodies. Importantly, most class II proteins elicited IFN-gamma responses that were significantly stronger than those produced by previously identified antigens. Among them, two class II proteins, ML0308 and ML2498, showed marked humoral and cellular immunogenicity, therefore providing promising candidates for the diagnosis of both tuberculoid and lepromatous forms of leprosy. PMID- 16368972 TI - Role in virulence of a Brucella abortus protein exhibiting lectin-like activity. AB - Brucella abortus is a facultative, intracellular zoonotic pathogen which can cause undulant fever in humans and abortions in cattle. A 14-kDa protein of B. abortus was previously identified to be immunogenic in animals infected with Brucella spp. In this study, we discovered that the 14-kDa protein possessed immunoglobulin binding and hemagglutination properties that appeared to be based on the protein's lectin-like properties. Hemagglutination inhibition experiments suggested that the 14-kDa protein has affinity towards mannose. Disruption of the gene encoding the 14-kDa protein in virulent B. abortus strain 2308 induced a rough-like phenotype with an altered smooth lipopolysaccharide (LPS) immunoblot profile and a significant reduction in the bacterium's ability to replicate in mouse spleens. However, the mutant strain was stably maintained in mouse spleens at 2.0 to 2.6 log(10) CFU/spleen from day 1 to week 6 after intraperitoneal inoculation with 4.65 log(10) CFU. In contrast to the case for the smooth virulent strain 2308, in the rough attenuated strain RB51 disruption of the 14 kDa protein's gene had no effect on the mouse clearance pattern. These findings indicate that the 14-kDa protein of B. abortus possesses lectin-like properties and is essential for the virulence of the species, probably because of its direct or indirect role in the synthesis of smooth LPS. PMID- 16368973 TI - Enterocyte cytoskeleton changes are crucial for enhanced translocation of nonpathogenic Escherichia coli across metabolically stressed gut epithelia. AB - Substantial data implicate the commensal flora as triggers for the initiation of enteric inflammation or inflammatory disease relapse. We have shown that enteric epithelia under metabolic stress respond to nonpathogenic bacteria by increases in epithelial paracellular permeability and bacterial translocation. Here we assessed the structural basis of these findings. Confluent filter-grown monolayers of the human colonic T84 epithelial cell line were treated with 0.1 mM dinitrophenol (which uncouples oxidative phosphorylation) and noninvasive, nonpathogenic Escherichia coli (strain HB101, 10(6) CFU) with or without pretreatment with various pharmacological agents. At 24 h later, apoptosis, tight junction protein expression, transepithelial resistance (TER; a marker of paracellular permeability), and bacterial internalization and translocation were assessed. Treatment with stabilizers of microtubules (i.e., colchicine), microfilaments (i.e., jasplakinolide) and clathrin-coated pit endocytosis (i.e., phenylarsine oxide) all failed to block DNP+E. coli HB101-induced reductions in TER but effectively prevented bacterial internalization and translocation. Neither the TER defect nor the enhanced bacterial translocations were a consequence of increased apoptosis. These data show that epithelial paracellular and transcellular (i.e., bacterial internalization) permeation pathways are controlled by different mechanisms. Thus, epithelia under metabolic stress increase their endocytotic activity that can result in a microtubule-, microfilament-dependent internalization and transcytosis of bacteria. We speculate that similar events in vivo would allow excess unprocessed antigen and bacteria into the mucosa and could evoke an inflammatory response by, for example, the activation of resident or recruited immune cells. PMID- 16368974 TI - Members of the 30- to 32-kilodalton mycolyl transferase family (Ag85) from culture filtrate of Mycobacterium avium subsp. paratuberculosis are immunodominant Th1-type antigens recognized early upon infection in mice and cattle. AB - The characterization of protective antigens is essential for the development of an effective, subunit-based vaccine against paratuberculosis. Surface-exposed and secreted antigens, present abundantly in mycobacterial culture filtrate (CF), are among the well-known protective antigens of Mycobacterium tuberculosis and Mycobacterium bovis. Culture filtrate, prepared from Mycobacterium avium subsp. paratuberculosis ATCC 19698 grown as a surface pellicle on synthetic Sauton medium, was strongly and early recognized in experimentally infected B6 bg/bg beige mice and cattle, as indicated by elevated spleen cell gamma interferon (IFN gamma) secretion and lymphoproliferative responses of peripheral blood mononuclear cells, respectively. Strong proliferative and ex vivo IFN-gamma responses against antigen 85 (Ag85) complex (a major protein component from M. bovis BCG culture filtrate) could be detected in cattle as early as 10 weeks after oral M. avium subsp. paratuberculosis infection. Synthetic peptides from the Ag85A and Ag85B components of this complex were strongly recognized, whereas T-cell responses were weaker against peptides from the Ag85C protein. A promiscuous T-cell epitope spanning amino acids 145 to 162 of Ag85B (identical sequence in M. bovis and M. avium subsp. paratuberculosis) was identified in experimentally infected cattle. Finally, young calves, born from cows with confirmed paratuberculosis, demonstrated proliferative responses to purified, recombinant Ag85A and Ag85B from M. avium subsp. paratuberculosis. These results indicate that the M. avium subsp. paratuberculosis Ag85 homologues are immunodominant T-cell antigens that are recognized early in experimental and natural infection of cattle. PMID- 16368975 TI - Production and characterization of guinea pig recombinant gamma interferon and its effect on macrophage activation. AB - Gamma interferon (IFN-gamma) plays a critical role in the protective immune responses against mycobacteria. We previously cloned a cDNA coding for guinea pig IFN-gamma (gpIFN-gamma) and reported that BCG vaccination induced a significant increase in the IFN-gamma mRNA expression in guinea pig cells in response to living mycobacteria and that the virulent H37Rv strain of Mycobacterium tuberculosis stimulated less IFN-gamma mRNA than did the attenuated H37Ra strain. In this study, we successfully expressed and characterized recombinant gpIFN gamma with a histidine tag at the N terminus (His-tagged rgpIFN-gamma) in Escherichia coli. rgpIFN-gamma was identified as an 18-kDa band in the insoluble fraction; therefore, the protein was purified under denaturing conditions and renatured. N-terminal amino acid sequencing of the recombinant protein yielded the sequence corresponding to the N terminus of His-tagged gpIFN-gamma. The recombinant protein upregulated major histocompatibility complex class II expression in peritoneal macrophages. The antiviral activity of rgpIFN-gamma was demonstrated with a guinea pig fibroblast cell line (104C1) infected with encephalomyocarditis virus. Interestingly, peritoneal macrophages treated with rgpIFN-gamma did not produce any nitric oxide but did produce hydrogen peroxide and suppressed the intracellular growth of mycobacteria. Furthermore, rgpIFN gamma induced morphological alterations in cultured macrophages. Thus, biologically active rgpIFN-gamma has been successfully produced and characterized in our laboratory. The study of rgpIFN-gamma will further increase our understanding of the cellular and molecular responses induced by BCG vaccination in the guinea pig model of pulmonary tuberculosis. PMID- 16368976 TI - Comparison of gamma interferon-mediated antichlamydial defense mechanisms in human and mouse cells. AB - Gamma interferon (IFN-gamma)-induced effector mechanisms have potent antichlamydial activities that are critical to host defense. The most prominent and well-studied effectors are indoleamine dioxygenase (IDO) and nitric oxide (NO) synthase. The relative contributions of these mechanisms as inhibitors of chlamydial in vitro growth have been extensively studied using different host cells, induction mechanisms, and chlamydial strains with conflicting results. Here, we have undertaken a comparative analysis of cytokine- and lipopolysaccharide (LPS)-induced IDO and NO using an extensive assortment of human and murine host cells infected with human and murine chlamydial strains. Following cytokine (IFN-gamma or tumor necrosis factor alpha) and/or LPS treatment, the majority of human cell lines induced IDO but failed to produce NO. Conversely, the majority of mouse cell lines studied produced NO, not IDO. Induction of IDO in human cell lines inhibited growth of L2 and mouse pneumonitis agent, now referred to as Chlamydia muridarum MoPn equally in all but two lines, and inhibition was completely reversible by the addition of tryptophan. IFN-gamma treatment of mouse cell lines resulted in substantially greater reduction of L2 than MoPn growth. However, despite elevated NO production by murine cells, blockage of NO synthesis with the l-arginine analogue N-monomethyl-l-arginine only partially rescued chlamydial growth, suggesting the presence of another IFN gamma-inducible antichlamydial mechanism unique to murine cells. Moreover, NO generated from the chemical nitric oxide donor sodium nitroprusside showed little direct effect on chlamydial infectivity or growth, indicating a natural resistance to NO. Finally, IFN-gamma-inducible IDO expression in human HeLa cells was inhibited following exogenous NO treatment, resulting in a permissive environment for chlamydial growth. In summary, cytokine- and LPS-inducible effectors produced by human and mouse cells differ and, importantly, these host specific effector responses result in chlamydial strain-specific antimicrobial activities. PMID- 16368977 TI - Identification of a new spore wall protein from Encephalitozoon cuniculi. AB - Microsporidia form environmentally resistant spores that are critical for their host-to-host transmission and persistence in the environment. The spore walls of these organisms are composed of two layers, the exospore and the endospore. Two spore wall proteins (SWP1 and SWP2) have been previously identified in members of the Encephalitozoonidae family. These proteins localize to the exospore. The endospore is known to contain chitin, and a putative glycosylphosphatidylinositol (GPI)-anchored chitin deacetylase has been localized to the plasmalemma-endospore interface. Using proteomic techniques, we have identified a new spore wall protein (SWP3) that is located in the endospore. The gene for this protein is located on chromosome 1 and corresponds to the open reading frame ECU01_1270. SWP3 is predicted to have a signal peptide and to be GPI anchored. Consistent with these modifications, two-dimensional electrophoresis demonstrated that SWP3 has an acidic pI and a molecular mass of <20 kDa. By immunoelectron microscopy, this protein was found on the cell surface during sporogony and in the endospore in mature spores. SWP3 has several potential O-glycosylation sites, and it is possible that it is a mannosylated protein like the major polar tube protein (PTP1). PMID- 16368978 TI - Implications for induction of autoimmunity via activation of B-1 cells by Helicobacter pylori urease. AB - Besides various gastroduodenal diseases, Helicobacter pylori infection may be involved in autoimmune disorders like rheumatoid arthritis (RA) or idiopathic thrombocytopenic purpura. Such autoimmune disorders are often associated with autoreactive antibodies produced by B-1 cells, a subpopulation of B lymphocytes. These B-1 cells are mainly located in the pleural cavity or mucosal compartment. The existence of H. pylori urease-specific immunoglobulin A (IgA)-producing B cells in the mucosal compartment and of their specific IgM in the sera of acutely infected volunteers suggests the possibility that urease stimulates mucosal innate immune responses. Here, we show for the first time that purified H. pylori urease predominantly stimulates the B-1-cell population rather than B-2 cells, which produce antigen-specific conventional antibodies among splenic B220(+) B cells. The fact that such stimulation of B-1 cells was not affected by the addition of polymyxin B indicates that the effect of purified H. pylori urease was not due to the contamination with bacterial lipopolysaccharide. Furthermore, the production of various B-1-cell-related autoreactive antibodies such as IgM type rheumatoid factor, anti-single-stranded DNA antibody, and anti-phosphatidyl choline antibody was observed when the splenic B cells were stimulated with purified H. pylori urease in vitro. These findings suggest that H. pylori components, urease in particular, may be among the environmental triggers that initiate various autoimmune diseases via producing autoreactive antibodies through the activation of B-1 cells. The findings shown here offer important new insights into the pathogenesis of autoimmune disorders related to H. pylori infection. PMID- 16368979 TI - Target antigen, age, and duration of antigen exposure independently regulate immunoglobulin G subclass switching in malaria. AB - The isotype/subclass of immunoglobulin determines antibody function, but rather little is known about factors that direct class switching in vivo. To evaluate factors that might influence the maturation of the antibody response during infection, we conducted a seroepidemiological study of the immunoglobulin G (IgG) subclass response to four merozoite-associated antigens of Plasmodium falciparum in a mountainous region of northeastern Tanzania, where malaria endemicity declines with increasing altitudes. We found that IgG1/IgG3 class switching is independently affected by the nature of the antigen, cumulative exposure to the antigen, and the maturity of the immune system (i.e., the age of the individual). These observations provide insights into the effects of immune system maturity, the duration and intensity of antigen exposure, and inherent characteristics of individual antigens on the process of class switching in human B cells. Our data also throw light on the consequences of class switch decisions on the gradual acquisition of antimalarial immunity. PMID- 16368980 TI - The lpf gene cluster for long polar fimbriae is not involved in adherence of enteropathogenic Escherichia coli or virulence of Citrobacter rodentium. AB - Using the enteropathogenic Escherichia coli (EPEC) genome sequence, we found that EPEC E2348/69 has an lpfABCDE gene cluster homologous (about 60% identical at the protein level) to the Salmonella long polar fimbria (LPF) operon. To determine whether this operon is essential for adherence, the lpfABCD(E2)(3) genes were deleted from EPEC strain E2348/69 by allelic exchange. Analysis of the resulting EPECDeltalpfABCD(E23) strain showed no change in adherence to HeLa cells or to human intestinal biopsy cells in the in vitro organ culture (IVOC) system compared to the wild type. Sera from volunteers experimentally infected with E2348/69 showed no antibody response to the major subunit protein, LpfA. These results suggested that the lpf(E23) gene cluster is not necessary for EPEC adherence and attaching/effacing (A/E) lesion formation on human biopsy samples and is not expressed during human infection. We also identified an lpf gene cluster in Citrobacter rodentium strain ICC168 (lpf(cr)). A DeltalpfA(cr) mutant of ICC168 retained wild-type adherence and A/E lesion-forming activity on HeLa cells. C3H/HeJ mice were infected with a wild-type C. rodentium strain and its lpfA(cr) isogenic mutant. Both strains were recovered at high levels in stools, and there were no significant differences between the groups both in terms of the number of CFU/organ (colon and cecum) and in terms of the amount of hyperplasia, as measured by weight. Similar results were observed in a second mouse strain, C57BL/6. These data suggest that in addition to playing no apparent role in EPEC pathogenesis, lpf(cr) is not required for C. rodentium virulence in either the C3H/HeJ or C57BL/6 mouse model. PMID- 16368981 TI - Interaction with CagF is required for translocation of CagA into the host via the Helicobacter pylori type IV secretion system. AB - Development of severe gastric diseases is strongly associated with those strains of Helicobacter pylori that contain the cag pathogenicity island (PAI) inserted into the chromosome. The cag PAI encodes a type IV secretion system that translocates the major disease-associated virulence protein, CagA, into the host epithelial cell. CagA then affects host signaling pathways, leading to cell elongations and inflammation. Since the precise mechanism by which the CagA toxin is translocated by the type IV secretion system remained elusive, we used fusion proteins and immunoprecipitation studies to identify CagA-interacting secretion components. Here we demonstrate that CagA, in addition to other yet-unidentified proteins, interacts with CagF, presumably at the inner bacterial membrane. This interaction is required for CagA translocation, since an isogenic nonpolar cagF mutant was translocation deficient. Our results suggest that CagF may be a protein with unique chaperone-like function that is involved in the early steps of CagA recognition and delivery into the type IV secretion channel. PMID- 16368982 TI - Differential regulation of cytokine production by CD1d-restricted NKT cells in response to superantigen staphylococcal enterotoxin B exposure. AB - NKT cells are a heterogeneous population characterized by the ability to rapidly produce cytokines, such as interleukin 2 (IL-2), IL-4, and gamma interferon (IFN gamma) in response to infections by viruses, bacteria, and parasites. The bacterial superantigen staphylococcal enterotoxin B (SEB) interacts with T cells bearing the Vbeta3, -7, or -8 T-cell receptors, inducing their expansion and cytokine secretion, leading to death in some cases due to cytokine poisoning. The majority of NKT cells bear the Vbeta7 or -8 T-cell receptor, suggesting that they may play a role in regulating this response. Using mice lacking NKT cells (CD1d( /-) and Jalpha18(-/-) mice), we set out to identify the role of these cells in T cell expansion, cytokine secretion, and toxicity induced by exposure to SEB. We find that Vbeta8(+) CD4(+) T-cell populations similarly expand in wild-type (WT) and NKT cell-null mice and that NKT cells did not regulate the secretion of IL-2. By contrast, these cells positively regulated the secretion of IL-4 and IFN-gamma production and negatively regulated the secretion of tumor necrosis factor alpha (TNF-alpha). However, this negative regulation of TNF-alpha secretion by NKT cells provides only a minor protective effect on SEB-mediated shock in WT mice compared to mice lacking NKT cells. These data suggest that NKT cells may regulate the nature of the cytokine response to exposure to the superantigen SEB and may act as regulatory T cells during exposure to this superantigen. PMID- 16368983 TI - Dietary iron content mediates hookworm pathogenesis in vivo. AB - Hookworm infection is associated with growth delay and iron deficiency anemia in developing countries. A series of experiments were designed in order to test the hypothesis that host dietary iron restriction mediates susceptibility to hookworm infection using the hamster model of Ancylostoma ceylanicum. Animals were maintained on diets containing either 10 ppm iron (iron restricted) or 200 ppm iron (standard/high iron), followed by infection with A. ceylanicum third-stage larvae. Infected animals fed the standard diet exhibited statistically significant growth delay and reduced blood hemoglobin levels compared to uninfected controls on day 20 postinfection. In contrast, no statistically significant differences in weight or hemoglobin concentration were observed between infected and uninfected animals fed the iron-restricted diet. Moreover, iron-restricted animals were observed to have reduced intestinal worm burdens on day 10 and day 20 postinfection compared to those of animals maintained on the standard/high-iron diet. In a subsequent study, animals equilibrated on diets containing a range of iron levels (10 ppm, 40 ppm, 100 ppm, or 200 ppm) were infected with A. ceylanicum and followed for evidence of hookworm disease. Infected animals from the intermediate-dietary iron (40- and 100-ppm) groups exhibited greater weight loss and anemia than those in the low (10-ppm)- or high (200-ppm)-iron diet groups. Mortality was also significantly higher in the intermediate-dietary-iron groups. These data suggest that severe dietary iron restriction impairs hookworm development in vivo but that moderate iron restriction enhances host susceptibility to severe disease. PMID- 16368984 TI - Identification of Borrelia burgdorferi outer surface proteins. AB - Several Borrelia burgdorferi outer surface proteins have been identified over the past decade that are up-regulated by temperature- and/or mammalian host-specific signals as this spirochete is transmitted from ticks to mammals. Given the potential role(s) that these differentially up-regulated proteins may play in B. burgdorferi transmission and Lyme disease pathogenesis, much attention has recently been placed on identifying additional borrelial outer surface proteins. To identify uncharacterized B. burgdorferi outer surface proteins, we previously performed a comprehensive gene expression profiling analysis of temperature shifted and mammalian host-adapted B. burgdorferi. The combined microarray analyses revealed that many genes encoding known and putative outer surface proteins are down-regulated in mammalian host-adapted B. burgdorferi. At the same time, however, several different genes encoding putative outer surface proteins were found to be up-regulated during the transmission and infection process. Among the putative outer surface proteins identified, biochemical and surface localization analyses confirmed that seven (Bb0405, Bb0689, BbA36, BbA64, BbA66, BbA69, and BbI42) are localized to the surface of B. burgdorferi. Furthermore, enzyme-linked immunosorbent assay analysis using serum from tick-infested baboons indicated that all seven outer surface proteins identified are immunogenic and that antibodies are generated against all seven during a natural infection. Specific antibodies generated against all seven of these surface proteins were found to be bactericidal against B. burgdorferi, indicating that these newly identified outer surface proteins are prime candidates for analysis as second generation Lyme disease vaccinogens. PMID- 16368985 TI - Identification of a surface protein of Streptococcus suis and evaluation of its immunogenic and protective capacity in pigs. AB - A Streptococcus suis surface protein reacting with convalescent-phase sera from pigs clinically infected by S. suis type 2 was identified. The apparent 110-kDa protein, designated Sao, exhibits typical features of membrane-anchored surface proteins of gram-positive bacteria, such as a signal sequence and an LPVTG membrane anchor motif. In spite of high identity with the partially sequenced genomes of S. suis Canadian strain 89/1591 and European strain P1/7, Sao does not share significant homology with other known sequences. However, a conserved avirulence domain that is often found in plant pathogens has been detected. Electron microscopy using an Sao-specific antiserum has confirmed the surface location of the Sao protein on S. suis. The Sao-specific antibody reacts with cell lysates of 28 of 33 S. suis serotypes and 25 of 26 serotype 2 isolates in immunoblots, suggesting its high conservation in S. suis species. The immunization of piglets with recombinant Sao elicits a significant humoral antibody response. However, the antibody response is not reflected in protection of pigs that are intratracheally challenged with a virulent strain in our conventional vaccination model. PMID- 16368986 TI - Immunogenicity and protection of a recombinant human adenovirus serotype 35-based malaria vaccine against Plasmodium yoelii in mice. AB - Given the promise of recombinant adenovirus type 5 (rAd5) as a malaria vaccine carrier in preclinical models, we evaluated the potency of rAd35 coding for Plasmodium yoelii circumsporozoite protein (rAd35PyCS). We chose rAd35 since a survey with serum samples from African subjects demonstrated that human Ad35 has a much lower seroprevalence of 20% and a much lower geometric mean neutralizing antibody titer (GMT) of 48 compared to Ad5 (seroprevalence, 85%; GMT, 1,261) in countries with a high malaria incidence. We also demonstrated that immunization with rAd35PyCS induced a dose-dependent and potent, CS-specific CD8(+) cellular and humoral immune response and conferred significant inhibition (92 to 94%) of liver infection upon high-dose sporozoite challenge. Furthermore, we showed that in mice carrying neutralizing antibody activity against Ad5, mimicking a human situation, CS-specific T- and B-cell responses were significantly dampened after rAd5PyCS vaccination, resulting in loss of inhibition of liver infection upon sporozoite challenge. In contrast, rAd35 vaccine was as potent in naive mice as in Ad5-preimmunized mice. Finally, we showed that heterologous rAd35-rAd5 prime boost regimens were more potent than rAd35-rAd35 because of induction of anti Ad35 antibodies after rAd35 priming. The latter data provide a further rationale for developing rAd prime-boost regimens but indicate that priming and boosting Ad vectors must be immunologically distinct and also should be distinct from Ad5. Collectively, the data presented warrant further development of rAd35-based vaccines against human malaria. PMID- 16368987 TI - Characterization, distribution, and expression of novel genes among eight clinical isolates of Streptococcus pneumoniae. AB - Eight low-passage-number Streptococcus pneumoniae clinical isolates, each of a different serotype and a different multilocus sequence type, were obtained from pediatric participants in a pneumococcal vaccine trial. Comparative genomic analyses were performed with these strains and two S. pneumoniae reference strains. Individual genomic libraries were constructed for each of the eight clinical isolates, with an average insert size of approximately 1 kb. A total of 73,728 clones were picked for arraying, providing more than four times genomic coverage per strain. A subset of 4,793 clones were sequenced, for which homology searches revealed that 750 (15.6%) of the sequences were unique with respect to the TIGR4 reference genome and 263 (5.5%) clones were unrelated to any available streptococcal sequence. Hypothetical translations of the open reading frames identified within these novel sequences showed homologies to a variety of proteins, including bacterial virulence factors not previously identified in S. pneumoniae. The distribution and expression patterns of 58 of these novel sequences among the eight clinical isolates were analyzed by PCR- and reverse transcriptase PCR-based analyses, respectively. These unique sequences were nonuniformly distributed among the eight isolates, and transcription of these genes in planktonic cultures was detected in 81% (172/212) of their genic occurrences. All 58 novel sequences were transcribed in one or more of the clinical strains, suggesting that they all correspond to functional genes. Sixty five percent (38/58) of these sequences were found in 50% or less of the clinical strains, indicating a significant degree of genomic plasticity among natural isolates. PMID- 16368988 TI - Identification of CsrC and characterization of its role in epithelial cell invasion in Salmonella enterica serovar Typhimurium. AB - The csr regulatory system of Salmonella regulates the expression of the genes of Salmonella pathogenicity island 1 (SPI1) required for the invasion of epithelial cells. This system consists of the posttranscriptional regulator CsrA and an untranslated regulatory RNA, CsrB, that opposes the action of CsrA. Here we identify and characterize the role of a second regulatory RNA, CsrC, whose ortholog was discovered previously in Escherichia coli. We show that a mutant of csrC has only mild defects in invasion and the expression of SPI1 genes, as does a mutant of csrB, but that a double csrB csrC mutant is markedly deficient in these properties, suggesting that the two regulatory RNAs play redundant roles in the control of invasion. We further show that CsrC, like CsrB, is controlled by the BarA/SirA two-component regulator but that a csrB csrC mutant exhibits a loss of invasion equivalent to that of a barA or sirA mutant, indicating that much of the effect of BarA/SirA on invasion functions through its control of CsrB and CsrC. In addition to their control by BarA/SirA, each regulatory RNA is also controlled by other components of the csr system. The loss of csrB was found to increase the level of CsrC by sevenfold, while the loss of csrC increased CsrB by nearly twofold. Similarly, the overexpression of csrA increased CsrC by nearly 11 fold and CsrB by 3-fold and also significantly increased the stability of both RNAs. PMID- 16368989 TI - Identification of differentially expressed genes in virulent and nonvirulent Entamoeba species: potential implications for amebic pathogenesis. AB - Entamoeba histolytica is a protozoan parasite that causes colitis and liver abscesses. Several Entamoeba species and strains with differing levels of virulence have been identified. E. histolytica HM-1:IMSS is a virulent strain, E. histolytica Rahman is a nonvirulent strain, and Entamoeba dispar is a nonvirulent species. We used an E. histolytica DNA microarray consisting of 2,110 genes to assess the transcriptional differences between these species/strains with the goal of identifying genes whose expression correlated with a virulence phenotype. We found 415 genes expressed at lower levels in E. dispar and 32 genes with lower expression in E. histolytica Rahman than in E. histolytica HM-1:IMSS. Overall, 29 genes had decreased expression in both the nonvirulent species/strains than the virulent E. histolytica HM-1:IMSS. Interestingly, a number of genes with potential roles in stress response and virulence had decreased expression in either one or both nonvirulent Entamoeba species/strains. These included genes encoding Fe hydrogenase (9.m00419), peroxiredoxin (176.m00112), type A flavoprotein (6.m00467), lysozyme (6.m00454), sphingomyelinase C (29.m00231), and a hypothetical protein with homology to both a Plasmodium sporozoite threonine asparagine-rich protein (STARP) and a streptococcal hemagglutinin (238.m00054). The function of these genes in Entamoeba and their specific roles in parasite virulence need to be determined. We also found that a number of the non-long terminal-repeat retrotransposons (EhLINEs and EhSINEs), which have been shown to modulate gene expression and genomic evolution, had lower expression in the nonvirulent species/strains than in E. histolytica HM-1:IMSS. Our results, identifying expression profiles and patterns indicative of a virulence phenotype, may be useful in characterizing the transcriptional framework of virulence. PMID- 16368990 TI - Involvement of potD in Streptococcus pneumoniae polyamine transport and pathogenesis. AB - Polyamines such as putrescine, spermidine, and cadaverine are small, polycationic molecules that are required for optimal growth in all cells. The intracellular concentrations of these molecules are maintained by de novo synthesis and transport pathways. The human pathogen Streptococcus pneumoniae possesses a putative polyamine transporter (pot) operon that consists of the four pot specific genes potABCD. The studies presented here examined the involvement of potD in polyamine transport and in pneumococcal pathogenesis. A potD-deficient mutant was created in the mouse-virulent serotype 3 strain WU2 by insertion duplication mutagenesis. The growth of the WU2DeltapotD mutant was identical to that of the wild-type strain WU2 in vitro in rich media. However, WU2DeltapotD possessed severely delayed growth compared to wild-type WU2 in the presence of the polyamine biosynthesis inhibitors DFMO (alpha-dimethyl-fluoroornitithine) and MGBG [methylgloxal-bis (guanyl hydrazone)]. The mutant strain also showed a significant attenuation in virulence within murine models of systemic and pulmonary infection regardless of the inoculation route or location. These data suggest that potD is involved in pneumococcal polyamine transport and is important for pathogenesis within various infection models. PMID- 16368991 TI - Human recombinant antimannan immunoglobulin G1 antibody confers resistance to hematogenously disseminated candidiasis in mice. AB - Mannan is a major cell wall component found in Candida species. Natural antimannan antibody is present in sera from most normal adults, but its role in host resistance to hematogenously disseminated candidiasis is unknown. The purpose of this study was to develop recombinant human antimannan antibody and to study its protective function. A phage Fab display combinatorial library containing Fab genes from bone marrow lymphocytes was screened with Candida albicans yeast cells and chemically purified mannan. One antimannan Fab, termed M1, was converted to a full-length immunoglobulin G1 antibody, M1g1, and M1g1 was produced in CHO cells. The M1g1 epitope was found in C. albicans serotypes A and B, Candida tropicalis, Candida guilliermondii, Candida glabrata, and Candida parapsilosis. Its expression was active at both 23 degrees C and 37 degrees C and uniform over the cell surface. BALB/c mice passively immunized with M1g1 were more resistant than control mice to a lethal hematogenous infection by C. albicans, as evidenced by extension of survival in an M1g1 dose-dependent manner (P, 0.08 to <0.001) and by reduction in number of infection foci and their size in the kidney. In vitro studies found that M1g1 promoted phagocytosis and phagocytic killing of C. albicans yeast cells by mouse peritoneal macrophages and was required for activation of the mouse complement cascade. Thus, human antimannan antibody may have a protective role in host resistance to systemic candidiasis. PMID- 16368993 TI - Glyceraldehyde-3-phosphate dehydrogenase of Paracoccidioides brasiliensis is a cell surface protein involved in fungal adhesion to extracellular matrix proteins and interaction with cells. AB - The pathogenic fungus Paracoccidioides brasiliensis causes paracoccidioidomycosis, a pulmonary mycosis acquired by inhalation of fungal airborne propagules, which may disseminate to several organs and tissues, leading to a severe form of the disease. Adhesion to and invasion of host cells are essential steps involved in the infection and dissemination of pathogens. Furthermore, pathogens use their surface molecules to bind to host extracellular matrix components to establish infection. Here, we report the characterization of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of P. brasiliensis as an adhesin, which can be related to fungus adhesion and invasion. The P. brasiliensis GAPDH was overexpressed in Escherichia coli, and polyclonal antibody against this protein was obtained. By immunoelectron microscopy and Western blot analysis, GAPDH was detected in the cytoplasm and the cell wall of the yeast phase of P. brasiliensis. The recombinant GAPDH was found to bind to fibronectin, laminin, and type I collagen in ligand far-Western blot assays. Of special note, the treatment of P. brasiliensis yeast cells with anti-GAPDH polyclonal antibody and the incubation of pneumocytes with the recombinant protein promoted inhibition of adherence and internalization of P. brasiliensis to those in vitro cultured cells. These observations indicate that the cell wall-associated form of the GAPDH in P. brasiliensis could be involved in mediating binding of fungal cells to fibronectin, type I collagen, and laminin, thus contributing to the adhesion of the microorganism to host tissues and to the dissemination of infection. PMID- 16368992 TI - T-helper-2 cytokine responses to Sj97 predict resistance to reinfection with Schistosoma japonicum. AB - Although schistosomiasis is effectively treated with Praziquantel, rapid reinfection with rebound morbidity precludes effective control based on chemotherapy alone and justifies current efforts to develop vaccines for these parasites. Using a longitudinal treatment-reinfection study design with 616 participants 7 to 30 years of age, we evaluated the relationship between cytokine responses to Schistosoma japonicum soluble adult worm extract (SWAP), Sj97, Sj22.6, and Sj67, measured 4 weeks after treatment with Praziquantel, and resistance to reinfection in a population from Leyte, The Philippines, where S. japonicum is endemic. S. japonicum transmission was high: 54.8% and 91.1% were reinfected within 6 and 18 months, respectively. A Th2 bias in the following cytokine ratios, interleukin-4 (IL-4)/IL-12, IL-5/IL-12, IL-13/IL-12, IL-4/gamma IFN (IFN-gamma), IL-5/IFN-gamma, and IL-13/IFN-gamma, in response to SWAP predicted a 1.4- to 2.9-month longer time to reinfection (P < 0.05) and a 27 to 55% lower intensity of reinfection (P < 0.05). Similarly, a Th2 bias in response to Sj97 predicted a 1.6- to 2.2-month longer time to reinfection (P < 0.05) and a 30 to 41% lower intensity of reinfection (P < 0.05). Only a high IL-5/IL-10 ratio in response to Sj22.6 predicted a 3.0-month-longer time to reinfection (P = 0.03). Cytokine responses to Sj67 were not associated with protection. In a large population-based treatment-reinfection study we found that Th2 responses to SWAP and Sj97 consistently predicted resistance to reinfection. These findings underscore Th2-type immune responses as central in human resistance to S. japonicum and support Sj97 as a leading vaccine candidate for this parasite. PMID- 16368994 TI - Complement and Fc function are required for optimal antibody prophylaxis against Pneumocystis carinii pneumonia. AB - Pneumocystis carinii is an opportunistic fungal pathogen that causes P. carinii pneumonia (PCP) in the immunocompromised host. We investigated the role of antibody Fc-mediated function in passive prophylaxis against the development of PCP in SCID mice. By comparison of anti-mouse P. carinii immunoglobulin G1 monoclonal antibody (MAb) 4F11(G1) and its F(ab')2 derivative in an intranasal immunoprophylaxis model, we determined that Fc-mediated function is required for maximum effect of this antibody. Comparison of efficacy of antibody prophylaxis in SCID mice depleted of complement to that in nondepleted mice demonstrated that complement fixation by MAb 4F11(G1) is also necessary for optimal effect of passively administered antibody, although residual protection was observed in complement-depleted SCID mice. The necessity of complement for optimal PCP prophylaxis by MAb 4F11(G1) suggests that complement may play a role in antibody mediated protection against development of PCP. PMID- 16368995 TI - Immunological correlates for protection against intranasal challenge of Bacillus anthracis spores conferred by a protective antigen-based vaccine in rabbits. AB - Correlates between immunological parameters and protection against Bacillus anthracis infection in animals vaccinated with protective antigen (PA)-based vaccines could provide surrogate markers to evaluate the putative protective efficiency of immunization in humans. In previous studies we demonstrated that neutralizing antibody levels serve as correlates for protection in guinea pigs (S. Reuveny et al., Infect. Immun. 69:2888-2893, 2001; H. Marcus et al., Infect. Immun. 72:3471-3477, 2004). In this study we evaluated similar correlates for protection by active and passive immunization of New Zealand White rabbits. Full immunization and partial immunization were achieved by single and multiple injections of standard and diluted doses of a PA-based vaccine. Passive immunization was carried out by injection of immune sera from rabbits vaccinated with PA-based vaccine prior to challenge with B. anthracis spores. Immunized rabbits were challenged by intranasal spore instillation with one of two virulent strains (strains Vollum and ATCC 6605). The immune competence was estimated by measuring the level of total anti-PA antibodies, the neutralizing antibody titers, and the conferred protective immunity. The results indicate that total anti-PA antibody titers greater than 1 x 10(5) conferred protection, whereas lower titers (between 10(4) and 10(5)) provided partial protection but failed to predict protection. Neutralizing antibody titers between 500 and 800 provided partial protection, while titers higher than 1,000 conferred protection. In conclusion, this study emphasizes that regardless of the immunization regimen or the time of challenge, neutralizing antibody titers are better predictors of protection than total anti-PA titers. PMID- 16368996 TI - Transcriptional activation of the Staphylococcus aureus putP gene by low-proline high osmotic conditions and during infection of murine and human tissues. AB - Staphylococcus aureus can grow virtually anywhere in the human body but needs to import proline through low- and high-affinity proline transporters to survive. This study examined the regulation of the S. aureus putP gene, which encodes a high-affinity proline permease. putP::lacZ and putP::lux transcriptional fusions were constructed and integrated into the genomes of several S. aureus strains. Enzyme activity was measured after growth in media with various osmolyte concentrations. As osmolarity rose, putP expression increased, with a plateau at 2 M for NaCl in strain LL3-1. Proline concentrations as low as 17.4 muM activated expression of the putP gene. The putP::lux fusion was also integrated into the genomes of S. aureus strains that were either SigB inactive (LL3-1, 8325-4, and SH1003) or SigB active (Newman and SH1000). SigB inactive strains showed increased putP gene expression as NaCl concentrations rose, whereas SigB active strains displayed a dramatic decrease in putP expression, suggesting that the alternative sigma factor B plays a negative role in putP regulation. Mice inoculated with S. aureus strains containing the putP::lux fusion exhibited up to a 715-fold increase in putP expression, although levels in the various murine organs differed. Moreover, urine from human patients infected with S. aureus showed elevated putP levels by use of a PCR procedure, whereas blood and some abscess material had no significant increase. Thus, putP is transcriptionally activated by a low-proline and high osmotic environment both in growth media and in murine or human clinical specimens. PMID- 16368997 TI - DNA adenine methyltransferase influences the virulence of Aeromonas hydrophila. AB - Among the various virulence factors produced by Aeromonas hydrophila, a type II secretion system (T2SS)-secreted cytotoxic enterotoxin (Act) and the T3SS are crucial in the pathogenesis of Aeromonas-associated infections. Our laboratory molecularly characterized both Act and the T3SS from a diarrheal isolate, SSU of A. hydrophila, and defined the role of some regulatory genes in modulating the biological effects of Act. In this study, we cloned, sequenced, and expressed the DNA adenine methyltransferase gene of A. hydrophila SSU (dam(AhSSU)) in a T7 promoter-based vector system using Escherichia coli ER2566 as a host strain, which could alter the virulence potential of A. hydrophila. Recombinant Dam, designated as M.AhySSUDam, was produced as a histidine-tagged fusion protein and purified from an E. coli cell lysate using nickel affinity chromatography. The purified Dam had methyltransferase activity, based on its ability to transfer a methyl group from S-adenosyl-l-methionine to N(6)-methyladenine-free lambda DNA and to protect methylated lambda DNA from digestion with DpnII but not against the DpnI restriction enzyme. The dam gene was essential for the viability of the bacterium, and overproduction of Dam in A. hydrophila SSU, using an arabinose inducible, P(BAD) promoter-based system, reduced the virulence of this pathogen. Specifically, overproduction of M.AhySSUDam decreased the motility of the bacterium by 58%. Likewise, the T3SS-associated cytotoxicity, as measured by the release of lactate dehydrogenase enzyme in murine macrophages infected with the Dam-overproducing strain, was diminished by 55% compared to that of a control A. hydrophila SSU strain harboring the pBAD vector alone. On the contrary, cytotoxic and hemolytic activities associated with Act as well as the protease activity in the culture supernatant of a Dam-overproducing strain were increased by 10-, 3-, and 2.4-fold, respectively, compared to those of the control A. hydrophila SSU strain. The Dam-overproducing strain was not lethal to mice (100% survival) when given by the intraperitoneal route at a dose twice that of the 50% lethal dose, which within 2 to 3 days killed 100% of the animals inoculated with the A. hydrophila control strain. Taken together, our data indicated alteration of A. hydrophila virulence by overproduction of Dam. PMID- 16368998 TI - GroEL of Lactobacillus johnsonii La1 (NCC 533) is cell surface associated: potential role in interactions with the host and the gastric pathogen Helicobacter pylori. AB - Heat shock proteins of the GroEL or Hsp60 class are highly conserved proteins essential to all living organisms. Even though GroEL proteins are classically considered intracellular proteins, they have been found at the surface of several mucosal pathogens and have been implicated in cell attachment and immune modulation. The purpose of the present study was to investigate the GroEL protein of a gram-positive probiotic bacterium, Lactobacillus johnsonii La1 (NCC 533). Its presence at the bacterial surface was demonstrated using a whole-cell enzyme linked immunosorbent assay and could be detected in bacterial spent culture medium by immunoblotting. To assess binding of La1 GroEL to mucins and intestinal epithelial cells, the La1 GroEL protein was expressed in Escherichia coli. We report here that La1 recombinant GroEL (rGroEL) binds to mucins and epithelial cells and that this binding is pH dependent. Immunomodulation studies showed that La1 rGroEL stimulates interleukin-8 secretion in macrophages and HT29 cells in a CD14-dependent mechanism. This property is common to rGroEL from other gram positive bacteria but not to the rGroEL of the gastric pathogen Helicobacter pylori. In addition, La1 rGroEL mediates the aggregation of H. pylori but not that of other intestinal pathogens. Our in vitro results suggest that GroEL proteins from La1 and other lactic acid bacteria might play a role in gastrointestinal homeostasis due to their ability to bind to components of the gastrointestinal mucosa and to aggregate H. pylori. PMID- 16368999 TI - Fibronectin binding protein BBK32 of the Lyme disease spirochete promotes bacterial attachment to glycosaminoglycans. AB - Borrelia burgdorferi, the agent of Lyme disease, causes a multisystemic illness that can affect the skin, heart, joints, and nervous system and is capable of attachment to diverse cell types. Among the host components recognized by this spirochete are fibronectin and glycosaminoglycans (GAGs). Three surface-localized GAG-binding bacterial ligands, Bgp, DbpA, and DbpB, have been previously identified, but recent studies suggested that at least one additional GAG-binding ligand is expressed on the spirochetal surface when the spirochete is adapted to the mammalian host environment. BBK32 is a surface lipoprotein that is produced during infection and that has been shown to bind to fibronectin. In this study, we show that, when BBK32 was produced from a shuttle vector in an otherwise nonadherent high-passage B. burgdorferi strain, the protein localized on the bacterial surface and conferred attachment to fibronectin and to mammalian cell monolayers. In addition, the high-passage strain producing BBK32 bound to purified preparations of the GAGs dermatan sulfate and heparin, as well as to these GAGs on the surfaces of cultured mammalian cells. Recombinant BBK32 recognized purified heparin, indicating that the bacterial attachment to GAGs was due to direct binding by BBK32. This GAG-binding activity of BBK32 is apparently independent of fibronectin recognition, because exogenous heparin had no effect on BBK32-mediated bacterial binding to fibronectin. PMID- 16369000 TI - Type IV pili of Neisseria gonorrhoeae influence the activation of human CD4+ T cells. AB - Neisseria gonorrhoeae is the causative agent of the sexually transmitted disease gonorrhea, and infection with this organism is typically associated with an intense inflammatory response. In many individuals, however, the infection is asymptomatic and can progress to serious secondary complications. The type IV pili of Neisseria gonorrhoeae mediate binding of the bacteria to host cells and are involved in cellular signal transduction. In these studies we have demonstrated that gonococcal pili influence human CD4+ T cells by using isogenic strains of N. gonorrhoeae with piliated and nonpiliated phenotypes. To determine the impact of piliation on the cellular status, we examined the expression of activation markers, cellular proliferation, and the production of cytokines after infection. The activation marker CD69 showed significantly increased expression on cells infected with the piliated strain, and this expression was dependent on costimulation of the T-cell receptor. Infection with piliated gonococci also altered T-cell proliferation and influenced the production of the regulatory cytokine interleukin-10. PilC, the putative pilus adhesin, was also observed to influence cellular activation but had no impact on the proliferation of cells further indicating that pilus-mediated adhesion is important in gonococcal stimulation of CD4+ T cells. These results show that the piliation status of gonococci influences CD4+ T-cell activation and that the adhesion mediated by pilus components aids in the regulation of the T-cell response to N. gonorrhoeae. PMID- 16369001 TI - Identification and characterization of the capsular polysaccharide (K-antigen) locus of Porphyromonas gingivalis. AB - Capsular polysaccharides of gram-negative bacteria play an important role in maintaining the structural integrity of the cell in hostile environments and, because of their diversity within a given species, can act as useful taxonomic aids. In order to characterize the genetic locus for capsule biosynthesis in the oral gram-negative bacterium Porphyromonas gingivalis, we analyzed the genome of P. gingivalis W83 which revealed two candidate loci at PG0106-PG0120 and PG1135 PG1142 with sufficient coding capacity and appropriate gene functions based on comparisons with capsule-coding loci in other bacteria. Insertion and deletion mutants were prepared at PG0106-PG0120 in P. gingivalis W50-a K1 serotype. Deletion of PG0109-PG0118 and PG0116-PG0120 both yielded mutants which no longer reacted with antisera to K1 serotypes. Restriction fragment length polymorphism analysis of the locus in strains representing all six K-antigen serotypes and K( ) strains demonstrated significant variation between serotypes and limited conservation within serotypes. In contrast, PG1135-PG1142 was highly conserved in this collection of strains. Sequence analysis of the capsule locus in strain 381 (K(-) strain) demonstrated synteny with the W83 locus but also significant differences including replacement of PG0109-PG0110 with three unique open reading frames, deletion of PG0112-PG0114, and an internal termination codon within PG0106, each of which could contribute to the absence of capsule expression in this strain. Analysis of the Arg-gingipains in the capsule mutants of strain W50 revealed no significant changes to the glycan modifications of these enzymes, which indicates that the glycosylation apparatus in P. gingivalis is independent of the capsule biosynthetic machinery. PMID- 16369002 TI - Effect of inactivation of the global oxidative stress regulator oxyR on the colonization ability of Escherichia coli O1:K1:H7 in a mouse model of ascending urinary tract infection. AB - To survive within the host urinary tract, Escherichia coli strains that cause urinary tract infection (UTI) presumably must overcome powerful oxidant stresses, including the oxygen-dependent killing mechanisms of neutrophils. Accordingly, we assessed the global oxygen stress regulator OxyR of Escherichia coli as a possible virulence factor in UTI by determining the impact of oxyR inactivation on experimental urovirulence in CBA/J and C57BL (both wild-type and p47(phox-/-)) mice. The oxyR and oxyS genes of wild-type E. coli strain Ec1a (O1:K1:H7) were replaced with a kanamycin resistance cassette to produce an oxyRS mutant. During in vitro growth in broth or human urine, the oxyRS mutant exhibited the same log phase growth rate (broth) and plateau density (broth and urine) as Ec1a, despite its prolonged lag phase (broth) or initial decrease in concentration (urine). The mutant, and oxyRS mutants of other wild-type ExPEC strains, exhibited significantly increased in vitro susceptibility to inhibition by H(2)O(2), which, like the altered growth kinetics observed with oxyRS inactivation, were reversed by restoration of oxyR on a multiple-copy-number plasmid. In CBA/J mice, Ec1a significantly outcompeted its oxyRS mutant (by >1 log(10)) in urine, bladder, and kidney cultures harvested 48 h after perurethral inoculation of mice, whereas an oxyR-complemented mutant exhibited equal or greater colonizing ability than that of the parent. Although C57BL mice were less susceptible to experimental UTI than CBA/J mice, wild-type and p47(phox-/-) C57BL mice were similarly susceptible, and the oxyR mutant of Ec1a was similarly attenuated in C57BL mice, regardless of the p47(phox) genotype, as in CBA/J mice. Within the E. coli Reference collection, 94% of strains were positive for oxyR. These findings fulfill the second and third of Koch's molecular postulates for oxyR as a candidate virulence facilitating factor in E. coli and indicate that oxyR is a broadly prevalent potential target for future preventive interventions against UTI due to E. coli. They also suggest that neutrophil phagocyte oxidase is not critical for defense against E. coli UTI and that the major oxidative stresses against which OxyR protects E. coli within the host milieu are not phagocyte derived. PMID- 16369003 TI - Roles of macrophages and neutrophils in the early host response to Bacillus anthracis spores in a mouse model of infection. AB - The development of new approaches to combat anthrax requires that the pathogenesis and host response to Bacillus anthracis spores be better understood. We investigated the roles that macrophages and neutrophils play in the progression of infection by B. anthracis in a mouse model. Mice were treated with a macrophage depletion agent (liposome-encapsulated clodronate) or with a neutrophil depletion agent (cyclophosphamide or the rat anti-mouse granulocyte monoclonal antibody RB6-8C5), and the animals were then infected intraperitoneally or by aerosol challenge with fully virulent, ungerminated B. anthracis strain Ames spores. The macrophage-depleted mice were significantly more susceptible to the ensuing infection than the saline-pretreated mice, whereas the differences observed between the neutropenic mice and the saline pretreated controls were generally not significant. We also found that augmenting peritoneal neutrophil populations before spore challenge did not increase resistance of the mice to infection. In addition, the bacterial load in macrophage-depleted mice was significantly greater and appeared significantly sooner than that observed with the saline-pretreated mice. However, the bacterial load in the neutropenic mice was comparable to that of the saline-pretreated mice. These data suggest that, in our model, neutrophils play a relatively minor role in the early host response to spores, whereas macrophages play a more dominant role in early host defenses against infection by B. anthracis spores. PMID- 16369004 TI - Two domains within the Mycoplasma hyopneumoniae cilium adhesin bind heparin. AB - Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia, a chronic and economically significant respiratory disease that affects swine production worldwide. M. hyopneumoniae adheres to and adversely affects the function of ciliated epithelial cells of the respiratory tract, and the cilium adhesin (Mhp183, P97) is intricately but not exclusively involved in this process. Although binding of pathogenic bacteria to glycosaminoglycans is a recognized step in pathogenesis, knowledge of glycosaminoglycan-binding proteins in M. hyopneumoniae is lacking. However, heparin and other sulfated polysaccharides are known to block the binding of M. hyopneumoniae to purified swine respiratory cilia. In this study, four regions within the cilium adhesin were examined for the ability to bind heparin. Cilium adhesin fragments comprising 653 amino acids of the N terminus and 301 amino acids of the C terminus (containing two repeat regions, R1 and R2) were cloned and expressed. These fragments bound heparin in a dose-dependent and saturable manner with physiologically significant binding affinities of 0.27 +/- 0.02 microM and 1.89 +/- 0.33 microM, respectively. Heparin binding of both fragments was strongly inhibited by the sulfated polysaccharides fucoidan and mucin but not by chondroitin sulfate B. When the C-terminal repeat regions R1 and R2 were cloned separately and expressed, heparin-binding activity was lost, suggesting that both regions are required for heparin binding. The ability of the cilium adhesin to bind heparin indicates that this molecule plays a multifunctional role in the adherence of M. hyopneumoniae to host respiratory surfaces and therefore has important implications with respect to the pathogenesis of this organism. PMID- 16369005 TI - Role of the luxS quorum-sensing system in biofilm formation and virulence of Staphylococcus epidermidis. AB - Nosocomial infections caused by Staphylococcus epidermidis are characterized by biofilm formation on implanted medical devices. Quorum-sensing regulation plays a major role in the biofilm development of many bacterial pathogens. Here, we describe luxS, a quorum-sensing system in staphylococci that has a significant impact on biofilm development and virulence. We constructed an isogenic DeltaluxS mutant strain of a biofilm-forming clinical isolate of S. epidermidis and demonstrated that luxS signaling is functional in S. epidermidis. The mutant strain showed increased biofilm formation in vitro and enhanced virulence in a rat model of biofilm-associated infection. Genetic complementation and addition of autoinducer 2-containing culture filtrate restored the wild-type phenotype, demonstrating that luxS repressed biofilm formation through a cell-cell signaling mechanism based on autoinducer 2 secretion. Enhanced production of the biofilm exopolysaccharide polysaccharide intercellular adhesin in the mutant strain is presumably the major cause of the observed phenotype. The agr quorum-sensing system has previously been shown to impact biofilm development and biofilm associated infection in a way similar to that of luxS, although by regulation of different factors. Our study indicates a general scheme of quorum-sensing regulation of biofilm development in staphylococci, which contrasts with that observed in many other bacterial pathogens. PMID- 16369006 TI - IdeS, a highly specific immunoglobulin G (IgG)-cleaving enzyme from Streptococcus pyogenes, is inhibited by specific IgG antibodies generated during infection. AB - IdeS, a recently discovered cysteine proteinase secreted by the important human pathogen Streptococcus pyogenes, interferes with phagocytic killing by specifically cleaving the heavy chain of immunoglobulin G. The fact that the enzyme targets one of the key molecules of the adapted immune response raised the question of whether an antibody response against IdeS could inhibit, i.e., neutralize, enzyme activity. Paired acute- and convalescent-phase serum samples from patients with pharyngotonsillitis (n = 10), bacteremia (n = 7), and erysipelas (n = 4) were analyzed. Antibodies with the ability to neutralize IdeS enzymatic activity were already found in two-thirds of acute-phase sera. However, patients who seroconverted to IdeS, in particular patients with pharyngotonsillitis and erysipelas, developed specific antibodies during convalescence with an increased capability to efficiently neutralize the enzymatic activity of IdeS. Also, the presence of neutralizing antibodies decreased the ability of IdeS to mediate bacterial survival in human immune blood. In patients with bacteremia, several acute-phase sera contained neutralizing antibodies, but no correlation was found to severity or outcome of invasive infections. Still, the fact that the human immune response targets the enzymatic activity of IdeS supports the view that the enzyme plays an important role during streptococcal infection. PMID- 16369007 TI - Urease produced by Coccidioides posadasii contributes to the virulence of this respiratory pathogen. AB - Urease activity during in vitro growth in the saprobic and parasitic phases of Coccidioides spp. is partly responsible for production of intracellular ammonia released into the culture media and contributes to alkalinity of the external microenvironment. Although the amino acid sequence of the urease of Coccidioides posadasii lacks a predicted signal peptide, the protein is transported from the cytosol into vesicles and the central vacuole of parasitic cells (spherules). Enzymatically active urease is released from the contents of mature spherules during the parasitic cycle endosporulation stage. The endospores, together with the urease and additional material which escape from the ruptured parasitic cells, elicit an intense host inflammatory response. Ammonia production by the spherules of C. posadasii is markedly increased by the availability of exogenous urea found in relatively high concentrations at sites of coccidioidal infection in the lungs of mice. Direct measurement of the pH at these infection sites revealed an alkaline microenvironment. Disruption of the urease gene of C. posadasii resulted in a marked reduction in the amount of ammonia secreted in vitro by the fungal cells. BALB/c mice challenged intranasally with the mutant strain showed increased survival, a well-organized granulomatous response to infection, and better clearance of the pathogen than animals challenged with either the parental or the reconstituted (revertant) strain. We conclude that ammonia and enzymatically active urease released from spherules during the parasitic cycle of C. posadasii contribute to host tissue damage, which exacerbates the severity of coccidioidal infection and enhances the virulence of this human respiratory pathogen. PMID- 16369008 TI - A recombinant aspartyl protease of Coccidioides posadasii induces protection against pulmonary coccidioidomycosis in mice. AB - Coccidioidomycosis is a respiratory disease of humans caused by the desert soil borne fungal pathogens Coccidioides spp. Recurrent epidemics of this mycosis in the southwestern United States have contributed significantly to escalated health care costs. Clinical and experimental studies indicate that prior symptomatic coccidioidomycosis induces immunity against subsequent infection, and activation of T cells is essential for containment of the pathogen and its clearance from host tissue. Development of a human vaccine against coccidioidomycosis has focused on recombinant T-cell-reactive antigens which elicit a durable protective immune response against pulmonary infection in mice. In this study we fractionated a protective multicomponent parasitic cell wall extract in an attempt to identify T-cell antigens. Immunoblots of electrophoretic separations of this extract identified patient seroreactive proteins which were subsequently excised from two-dimensional polyacrylamide gel electrophoresis gels, trypsin digested, and sequenced by tandem mass spectrometry. The full-length gene which encodes a dominant protein in the immunoblot was identified using established methods of bioinformatics. The gene was cloned and expressed, and the recombinant protein was shown to stimulate immune T cells in vitro. The deduced protein was predicted to contain epitopes that bind to human major histocompatibility complex class II molecules using a TEPITOPE-based algorithm. Synthetic peptides corresponding to the predicted T-cell epitopes induced gamma interferon production by immune T lymphocytes. The T-cell-reactive antigen, which is homologous to secreted fungal aspartyl proteases, protected mice against pulmonary infection with Coccidioides posadasii. We argue that this immunoproteomic/bioinformatic approach to the identification of candidate vaccines against coccidioidomycosis is both efficient and productive. PMID- 16369009 TI - CpG-oligodeoxynucleotide is a potent adjuvant with an Entamoeba histolytica Gal inhibitable lectin vaccine against amoebic liver abscess in gerbils. AB - The protozoan parasite Entamoeba histolytica causes invasive amoebiasis characterized by amoebic dysentery and liver abscesses (ALA). The E. histolytica galactose/N-acetyl-D-galactosamine-inhibitable lectin (Gal-lectin), an immunogenic surface molecule involved in colonization and invasion, is a promising vaccine candidate against amoebiasis. Gal-lectin is known to induce Th1 cytokines in macrophages and spleen cells in vitro, and a Th1 response is thought to be protective against ALA. In this study, we report the use of cytosine guanine oligodeoxynucleotide (CpG-ODN) as adjuvant to augment Th1 responses against Gal-lectin in the gerbil model of ALA. Gerbils were vaccinated intramuscularly with the native Gal-lectin plus CpG-ODN or a paired non-CpG control GpC-ODN, and control gerbils received CpG-ODN alone. One week after the last boost gerbils were challenged intrahepatically with 10(6) amoebae. Gerbils receiving CpG-ODN as adjuvant with Gal-lectin were completely protected against the development of ALA, whereas 50% of gerbils receiving GpC-ODN and Gal-lectin developed ALA and 85% of controls developed ALA. Stronger lymphoproliferation in response to the Gal-lectin and higher prechallenge titers of serum Gal-lectin specific antibodies, capable of blocking amoebic adherence, were observed when CpG-ODN was used as adjuvant. Gerbils vaccinated with CpG-ODN and Gal-lectin also had significantly higher levels of gamma interferon, interleukin-12 (IL-12), and IL-2 mRNA than controls. These data indicate that CpG-ODN can enhance the Th1 responses, which improve the protective effects of Gal-lectin. This is the first report of the use of CpG as a potent Th1 adjuvant with Gal-lectin to increase protection against ALA formation. PMID- 16369010 TI - The UDP N-acetylgalactosamine 4-epimerase gene is essential for mesophilic Aeromonas hydrophila serotype O34 virulence. AB - Mesophilic Aeromonas hydrophila strains of serotype O34 typically express smooth lipopolysaccharide (LPS) on their surface. A single mutation in the gene that codes for UDP N-acetylgalactosamine 4-epimerase (gne) confers the O(-) phenotype (LPS without O-antigen molecules) on a strain in serotypes O18 and O34, but not in serotypes O1 and O2. The gne gene is present in all the mesophilic Aeromonas strains tested. No changes were observed for the LPS core in a gne mutant from A. hydrophila strain AH-3 (serotype O34). O34 antigen LPS contains N acetylgalactosamine, while no such sugar residue forms part of the LPS core from A. hydrophila AH-3. Some of the pathogenic features of A. hydrophila AH-3 gne mutants are drastically reduced (serum resistance or adhesion to Hep-2 cells), and the gne mutants are less virulent for fish and mice compared to the wild-type strain. Strain AH-3, like other mesophilic Aeromonas strains, possess two kinds of flagella, and the absence of O34 antigen molecules by gne mutation in this strain reduced motility without any effect on the biogenesis of both polar and lateral flagella. The reintroduction of the single wild-type gne gene in the corresponding mutants completely restored the wild-type phenotype (presence of smooth LPS) independently of the O wild-type serotype, restored the virulence of the wild-type strain, and restored motility (either swimming or swarming). PMID- 16369011 TI - MyD88-dependent pathways mediate resistance to Cryptosporidium parvum infection in mice. AB - Cryptosporidium spp. cause diarrheal disease worldwide. Innate immune responses mediating resistance to this parasite are not completely understood. To determine whether MyD88-dependent pathways play a role in resistance to Cryptosporidium parvum, we compared the course of infection in MyD88(-/-) mice to that in their wild-type (WT) littermate controls. Three- to 4-week-old mice were infected with C. parvum, and infection was monitored by quantifying fecal oocyst shedding. Twelve days postinfection, the histology of the intestines was examined to quantify intestinal parasite burden and to determine if there were any pathological changes. Fecal oocyst shedding and intestinal parasite burden were significantly greater in MyD88(-/-) mice than in littermate controls. Nonetheless, both WT and MyD88(-/-) mice cleared the infection within 3 weeks. These results indicate that MyD88-dependent pathways are involved in mediating initial resistance to C. parvum. Since gamma interferon (IFN-gamma) is known to mediate resistance to C. parvum, we also studied infection in MyD88(-/-) mice and WT controls in which this cytokine was temporarily neutralized. Fecal oocyst shedding, as well as intestinal parasite burden, intestinal inflammation, and mortality, was significantly greater in MyD88(-/-) mice in which IFN-gamma was neutralized than in IFN-gamma-neutralized WT mice or in MyD88(-/-) mice in which this cytokine was active. These results suggest that MyD88 and IFN-gamma had an additive effect in conferring protection from C. parvum infection. While this study confirms the importance of IFN-gamma in conferring resistance to infection with C. parvum, it suggests that MyD88-mediated pathways also play a role in innate immunity to this parasite. PMID- 16369012 TI - Plasmid interleukin-23 (IL-23), but not plasmid IL-27, enhances the protective efficacy of a DNA vaccine against Mycobacterium tuberculosis infection. AB - Protection against intracellular pathogens such as Mycobacterium tuberculosis requires the development of Th1-like T-cell responses. This in turn is dependent on the pattern of cytokine produced from dendritic cells (DCs) after infection. Three heterodimeric cytokines, interleukin-12 (IL-12), IL-23, and IL-27, as well as IL-18, contribute to the differentiation and expansion of naive CD4(+) T cells. In this study we compared the effects of plasmids expressing both chains of IL-12, IL-23, or IL-27 as adjuvants for DNA immunization against M. tuberculosis infection. The genes encoding p19 and p40 chains of IL-23 or EBI3 and p28 chains of IL-27 were cloned on either side of a self-cleaving peptide from the FMDV2A protein. The secretion of functional cytokines from transfected cells was detected with bioassays. Supernatant from p2AIL-23-transfected cells induced the release of IL-17 from activated lymphocytes, confirming the presence of bioactive IL-23. Further, supernatant from p2AIL-27-transfected cells stimulated a significant increase in the proliferation of peptide-stimulated transgenic CD4(+) T cells. In initial experiments, M. tuberculosis infection of DCs was more potent at inducing IL-12 and IL-23 secretion than infection with the vaccine strain Mycobacterium bovis bacille Calmette-Guerin (BCG), and no significant upregulation of IL-27 was observed. Coimmunization of C57BL/6 mice with DNA expressing M. tuberculosis antigen 85B (Ag85B; DNA85B) and plasmids expressing IL-23 or IL-12 stimulated stronger Ag85B-specific T-cell proliferative and IFN-gamma responses than DNA85B alone, whereas the addition of p2AIL-27 had no effect. Interestingly, DNA85B codelivered with p2AIL-12, but not p2AIL-23, reduced the immunoglobulin G antibody response. Both p2AIL-23 and p2AIL-12, but not p2AIL-27, enhanced the protective efficacy of DNA85B against aerosol M. tuberculosis challenge. Therefore, both p2AIL-23 and p2AIL-12 are valuable as cytokine adjuvants for increasing the protective antituberculosis immunity induced by DNA vaccines. PMID- 16369013 TI - Identification of the insulin-like growth factor II receptor as a novel receptor for binding and invasion by Listeria monocytogenes. AB - The gram-positive bacterium Listeria monocytogenes causes a life-threatening disease known as listeriosis. The mechanism by which L. monocytogenes invades mammalian cells is not fully understood, but the processes involved may provide targets to prevent and treat listeriosis. Here, for the first time, we have identified the insulin-like growth factor II receptor (IGFIIR; also known as the cation-independent mannose 6-phosphate receptor (CI)M6PR or CD222) as a novel receptor for binding and invasion of Listeria species. Random peptide phage display was employed to select a peptide sequence by panning with immobilized L. monocytogenes cells; this peptide sequence corresponds to a sequence within the mannose 6-phosphate binding site of the IGFIIR. All Listeria spp. specifically bound the labeled peptide but not a control peptide, which was demonstrated using fluorescence spectrophotometry and fluorescence-activated cell sorting. Further evidence for binding of the receptor by L. monocytogenes and L. innocua was provided by affinity purification of the bovine IGFIIR from fetal calf serum by use of magnetic beads coated with cell preparations of Listeria spp. as affinity matrices. Adherence to and invasion of mammalian cells by L. monocytogenes was significantly inhibited by both the synthetic peptide and mannose 6-phosphate but not by appropriate controls. These observations indicate a role for the IGFIIR in the adherence and invasion of L. monocytogenes of mammalian cells, perhaps in combination with known mechanisms. Ligation of IGFIIR by L. monocytogenes may be a novel mechanism that contributes to the regulation of infectivity, possibly in combination with other mechanisms. PMID- 16369014 TI - The ompA gene in Chlamydia trachomatis differs in phylogeny and rate of evolution from other regions of the genome. AB - Strains of Chlamydia trachomatis are classified into serovars based on nucleotide sequence differences in ompA, the gene that encodes the major outer membrane protein. Phylogenetic characterization of strains based on ompA, however, results in serovar groupings that are inconsistent with the distinguishing features of C. trachomatis pathobiology, e.g., tissue tropisms and disease presentation. We have compared nucleotide sequences at multiple sites distributed around the chlamydial genome from 18 strains representing 16 serovars; sampled regions included genes encoding housekeeping enzymes (totaling 2,073 bp), intergenic noncoding segments (1,612 bp), and a gene encoding a second outer membrane protein (porB; 1,023 bp), with the ompA sequence (1,194 bp) used for reference. These comparative analyses revealed substantial variation in nucleotide substitution patterns among the sampled regions, with average pairwise sequence differences ranging from 0.15% for the housekeeping genes to 12.1% for ompA. Phylogenetic characterization of the sampled genomic sequences yielded a strongly supported tree that divides the strains into groupings consistent with C. trachomatis biology and which has a topology quite distinct from the ompA tree. This phylogenetic incongruity can be accounted for by recombination of the ompA gene between different genomic backgrounds. We found, however, no evidence of recombination within or between any of the sampled regions around the C. trachomatis genome apart from ompA. Parallel analysis of published sequence data on four members of the pmp gene family are consistent with the phylogenetic analyses reported here. PMID- 16369015 TI - Construction and immunological characterization of a novel nontoxic protective pneumolysin mutant for use in future pneumococcal vaccines. AB - Pneumolysin, the pore-forming toxin produced by Streptococcus pneumoniae, may have an application as an immunogenic carrier protein in future pneumococcal conjugate vaccines. Most of the 90 S. pneumoniae serotypes identified produce pneumolysin; therefore, this protein may confer non-serotype-specific protection against pneumococcal infections such as pneumonia, meningitis, and otitis media. However, as pneumolysin is highly toxic, a nontoxic form of pneumolysin would be a more desirable starting point in terms of vaccine production. Previous pneumolysin mutants have reduced activity but retain residual toxicity. We have found a single amino acid deletion that blocks pore formation, resulting in a form of pneumolysin that is unable to form large oligomeric ring structures. This mutant is nontoxic at concentrations greater than 1,000 times that of the native toxin. We have demonstrated that this mutant is as immunogenic as native pneumolysin without the associated effects such as production of the inflammatory mediators interleukin-6 and cytokine-induced neutrophil chemoattractant KC, damage to lung integrity, and hypothermia in mice. Vaccination with this mutant protects mice from challenge with S. pneumoniae. Incorporation of this mutant pneumolysin into current pneumococcal vaccines may increase their efficacy. PMID- 16369016 TI - Identification and characterization of a novel bacterial virulence factor that shares homology with mammalian Toll/interleukin-1 receptor family proteins. AB - Many important bacterial virulence factors act as mimics of mammalian proteins to subvert normal host cell processes. To identify bacterial protein mimics of components of the innate immune signaling pathway, we searched the bacterial genome database for proteins with homology to the Toll/interleukin-1 receptor (TIR) domain of the mammalian Toll-like receptors (TLRs) and their adaptor proteins. A previously uncharacterized gene, which we have named tlpA (for TIR like protein A), was identified in the Salmonella enterica serovar Enteritidis genome that is predicted to encode a protein resembling mammalian TIR domains, We show that overexpression of TlpA in mammalian cells suppresses the ability of mammalian TIR-containing proteins TLR4, IL-1 receptor, and MyD88 to induce the transactivation and DNA-binding activities of NF-kappaB, a downstream target of the TIR signaling pathway. In addition, TlpA mimics the previously characterized Salmonella virulence factor SipB in its ability to induce activation of caspase-1 in a mammalian cell transfection model. Disruption of the chromosomal tlpA gene rendered a virulent serovar Enteritidis strain defective in intracellular survival and IL-1beta secretion in a cell culture infection model using human THP1 macrophages. Bacteria with disrupted tlpA also displayed reduced lethality in mice, further confirming an important role for this factor in pathogenesis. Taken together, our findings demonstrate that the bacterial TIR-like protein TlpA is a novel prokaryotic modulator of NF-kappaB activity and IL-1beta secretion that contributes to serovar Enteritidis virulence. PMID- 16369017 TI - Iron and pH homeostasis intersect at the level of Fur regulation in the gastric pathogen Helicobacter pylori. AB - Helicobacter pylori persistently colonizes the stomach of the majority of the world's population and is a tremendous medical burden due to its causal role in diverse gastric maladies. Since the stomach is a constantly changing environment, successful colonization of H. pylori within this niche requires regulation of bacterial gene expression to cope with the environmental fluctuations. In H. pylori, the ferric uptake regulator (Fur) has been shown to play an intricate role in adaptation of the bacterium to two conditions known to oscillate within the gastric mucosa: iron limitation and low pH. To extend our knowledge of the process of regulation and adaptation in H. pylori, we show that Fur is required for efficient colonization of the Mongolian gerbil: the mutant strain exhibits a 100-fold increase in the 50% infectious dose, as well as a 100-fold defect in competitive colonization, when coinfected with wild-type bacteria. Furthermore, we used DNA microarrays to identify genes whose expression was altered in a Fur deficient strain. We show that the Fur regulon of H. pylori consists of approximately 30 genes, most of which have been previously annotated as acid stress associated. Finally, we investigate the role of Fur in acid-responsive modulation of gene expression and show that a large number of genes are aberrantly expressed in the Fur mutant specifically upon acid exposure. This fact likely explains the requirement for this regulator for growth and colonization in the stomach. PMID- 16369018 TI - The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine. AB - Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract. The prototype ABU E. coli strain 83972 was originally isolated from a girl who had carried it asymptomatically for 3 years. Deliberate colonization of UTI-susceptible individuals with E. coli 83972 has been used successfully as an alternative approach for the treatment of patients who are refractory to conventional therapy. Colonization with strain 83972 appears to prevent infection with UPEC strains in such patients despite the fact that this strain is unable to express the primary adhesins involved in UTI, viz. P and type 1 fimbriae. Here we investigated the growth characteristics of E. coli 83972 in human urine and show that it can outcompete a representative spectrum of UPEC strains for growth in urine. The unique ability of ABU E. coli 83972 to outcompete UPEC in urine was also demonstrated in a murine model of human UTI, confirming the selective advantage over UPEC in vivo. Comparison of global gene expression profiles of E. coli 83972 grown in lab medium and human urine revealed significant differences in expression levels in the two media; significant down-regulation of genes encoding virulence factors such as hemolysin, lipid A, and capsular polysaccharides was observed in cells grown in urine. Clearly, divergent abilities of ABU E. coli and UPEC to exploit human urine as a niche for persistence and survival suggest that these key differences may be exploited for preventative and/or therapeutic approaches. PMID- 16369019 TI - Microbial pattern recognition receptors mediate M-cell uptake of a gram-negative bacterium. AB - The receptors involved in the sampling of particulate microbial antigens by the gut are largely unknown. Here we demonstrate for the first time in an in vitro M cell model and in situ in isolated murine intestinal segments that the receptors TLR-4, PAF-R, and alpha5beta1 integrin are all involved in mediating bacterial uptake associated with transcytosis. The pattern of expression of TLR-4 and alpha5beta1 integrin differed between M cells and enterocytes. There was increased apical expression of TLR-4 in M-cell cultures, and it was present on the apical surface of murine M cells but not enterocytes in situ. In contrast, PAF-R was expressed equally by both cell types in vitro and was abundantly expressed throughout the intestinal epithelium. Inhibition of TLR-4 and PAF-R, but not TLR-2, reduced gram-negative bacterial uptake by both cell types, whereas inhibition of the apically expressed alpha5beta1 integrin significantly reduced the ability of M cells to translocate bacteria. Hence, the involvement of each receptor was dependent not only on differences in the level of receptor expression but the cellular localization. Using bacteria that had mutations that affected the bacterial lipooligosaccharide structure indicated that the oligosaccharide moiety was important in bacterial uptake. Taken together, the data suggest that pathogen-associated molecular pattern interactions with pattern recognition receptors are key factors in M-cell recognition of intestinal antigens for mucosal immune priming. PMID- 16369020 TI - Virulence of broad- and narrow-host-range Salmonella enterica serovars in the streptomycin-pretreated mouse model. AB - Salmonella enterica subspecies I serovars are common bacterial pathogens causing diseases ranging from enterocolitis to systemic infections. Some serovars are adapted to specific hosts, whereas others have a broad host range. The molecular mechanisms defining the virulence characteristics and the host range of a given S. enterica serovar are unknown. Streptomycin pretreated mice provide a surrogate host model for studying molecular aspects of the intestinal inflammation (colitis) caused by serovar Typhimurium (S. Hapfelmeier and W. D. Hardt, Trends Microbiol. 13:497-503, 2005). Here, we studied whether this animal model is also useful for studying other S. enterica subspecies I serovars. All three tested strains of the broad-host-range serovar Enteritidis (125109, 5496/98, and 832/99) caused pronounced colitis and systemic infection in streptomycin pretreated mice. Different levels of virulence were observed among three tested strains of the host-adapted serovar Dublin (SARB13, SD2229, and SD3246). Several strains of host restricted serovars were also studied. Two serovar Pullorum strains (X3543 and 449/87) caused intermediate levels of colitis. No intestinal inflammation was observed upon infection with three different serovar Paratyphi A strains (SARB42, 2804/96, and 5314/98) and one serovar Gallinarum strain (X3796). A second serovar Gallinarum strain (287/91) was highly virulent and caused severe colitis. This strain awaits future analysis. In conclusion, the streptomycin pretreated mouse model can provide an additional tool to study virulence factors (i.e., those involved in enteropathogenesis) of various S. enterica subspecies I serovars. Five of these strains (125109, 2229, 287/91, 449/87, and SARB42) are subject of Salmonella genome sequencing projects. The streptomycin pretreated mouse model may be useful for testing hypotheses derived from this genomic data. PMID- 16369021 TI - Platelets potentiate brain endothelial alterations induced by Plasmodium falciparum. AB - Brain lesions of cerebral malaria (CM) are characterized by a sequestration of Plasmodium falciparum-parasitized red blood cells (PRBC) and platelets within brain microvessels, as well as by blood-brain barrier (BBB) disruption. In the present study, we evaluated the possibility that PRBC and platelets induce functional alterations in brain endothelium. In a human brain endothelial cell line, named HBEC-5i, exhibiting most of the features demanded for a pathophysiological study of BBB, tumor necrosis factor (TNF) or lymphotoxin alpha (LT-alpha) reduced transendothelial electrical resistance (TEER), enhanced the permeability to 70-kDa dextran, and increased the release of microparticles, a recently described indicator of disease severity in CM patients. In vitro cocultures showed that platelets or PRBC can have a direct cytotoxic effect on activated, but not on resting, HBEC-5i cells. Platelet binding was required, as platelet supernatant had no effect. Furthermore, platelets potentiated the cytotoxicity of PRBC for TNF- or LT-alpha-activated HBEC-5i cells when they were added prior to these cells on the endothelial monolayers. This effect was not observed when platelets were added after PRBC. Both permeability and TEER were strongly affected, and the apoptosis rate of HBEC-5i cells was dramatically increased. These findings provide insights into the mechanisms by which platelets can be deleterious to the brain endothelium during CM. PMID- 16369022 TI - Fusobacterium nucleatum transports noninvasive Streptococcus cristatus into human epithelial cells. AB - Analysis of human buccal epithelial cells frequently reveals an intracellular polymicrobial consortium of bacteria. Although several oral bacteria have been demonstrated to invade cultured epithelial cells, several others appear unable to internalize. We hypothesized that normally noninvasive bacteria may gain entry into epithelial cells via adhesion to invasive bacteria. Fusobacterium nucleatum is capable of binding to and invading oral epithelial cells. By contrast, Streptococcus cristatus binds weakly to host cells and is not internalized. F. nucleatum and S. cristatus coaggregate strongly via an arginine-sensitive interaction. Coincubation of KB or TERT-2 epithelial cells with equal numbers of F. nucleatum and S. cristatus bacteria led to significantly increased numbers of adherent and internalized streptococci. F. nucleatum also promoted invasion of KB cells by other oral streptococci and Actinomyces naeslundii. Dissection of fusobacterial or streptococcal adhesive interactions by using sugars, amino acids, or antibodies demonstrated that this phenomenon is due to direct attachment of S. cristatus to adherent and invading F. nucleatum. Inhibition of F. nucleatum host cell attachment and invasion with galactose, or fusobacterial streptococcal coaggregation by the arginine homologue l-canavanine, abrogated the increased S. cristatus adhesion to, and invasion of, host cells. In addition, polyclonal antibodies to F. nucleatum, which inhibited fusobacterial attachment to both KB cells and S. cristatus, significantly decreased invasion by both species. Similar decreases were obtained when epithelial cells were pretreated with cytochalasin D, staurosporine, or cycloheximide. These studies indicate that F. nucleatum may facilitate the colonization of epithelial cells by bacteria unable to adhere or invade directly. PMID- 16369023 TI - Invasion of the placenta during murine listeriosis. AB - Feto-placental infections due to Listeria monocytogenes represent a major threat during pregnancy, and the underlying mechanisms of placental invasion remain poorly understood. Here we used a murine model of listeriosis (pregnant mice, infected at day 14 of gestation) to investigate how this pathogen invades and grows within the placenta to ultimately infect the fetus. When L. monocytogenes is injected intravenously, the invasion of the placenta occurs early after the initial bacteremia, allowing the placental growth of the bacteria, which is an absolute requirement for vertical transmission to the fetus. Kinetically, bacteria first target the cells lining the central arterial canal of the placenta, which stain positively with cytokeratin, demonstrating their fetal trophoblast origin. Bacteria then disseminate rapidly to the other trophoblastic structures, like syncytiotrophoblast cells lining the villous core in the labyrinthine zone of placenta. Additionally, we found that an inflammatory reaction predominantly constituted of polymorphonuclear cells occurs in the villous placenta and participates in the control of infection. Altogether, our results suggest that the infection of murine placenta is dependent, at the early phase, on circulating bacteria and their interaction with endovascular trophoblastic cells. Subsequently, the bacteria spread to the other trophoblastic cells before crossing the placental barrier. PMID- 16369024 TI - CD4+-T-cell effector functions and costimulatory requirements essential for surviving mucosal infection with Citrobacter rodentium. AB - Citrobacter rodentium causes an attaching and effacing infection of the mouse colon. Surprisingly, protective adaptive immunity against this mucosal pathogen requires a systemic T-cell-dependent antibody response. To define CD4+ T-cell effector functions promoting this systemic defense of infected epithelial surfaces, studies were undertaken in weaning-age mice lacking costimulatory molecules CD28 or CD40L or cytokines gamma interferon (IFN-gamma) or interleukin 4 (IL-4). Adoptive transfer of CD4+ T cells from wild-type, CD28(-/-), CD40L(-/ ), or IFN-gamma(-/-) donors to CD4(-/-) recipients delineated functions of these CD4+ T-cell-expressed molecules on the outcome of infection. Wild-type and IL-4( /-) mice successfully resolved infection, while 70% of IFN-gamma(-/-) mice survived. In contrast, all CD28(-/-) mice succumbed during acute infection. While fewer than half of CD40L(-/-) mice succumbed acutely, surviving mice failed to clear infection, resulting in progressive mucosal destruction, polymicrobial sepsis, and death 1 to 2 weeks later than in CD28(-/-) mice. Downstream of CD28 mediated effects, CD4+ T-cell-expressed CD40L proved essential for generating acute pathogen-specific immunoglobulin M (IgM) and early IgG, which reduced pathogen burdens. However, deficiency of CD4+ T-cell-expressed IFN-gamma did not adversely impact survival or development of protective antibody in adoptively transferred CD4(-/-) recipients, though it impacted Th1 antibody responses. These findings demonstrate that CD4+ T-cell-expressed CD40L promotes the rapid production of protective systemic antibody during acute infection, while deficiencies of IL-4 or of CD4+ T-cell-expressed IFN-gamma can be overcome. These findings have important implications for understanding the role of T-helper-cell responses during infections involving mucosal surfaces. PMID- 16369025 TI - Genome engineering in Bacillus anthracis using Cre recombinase. AB - Genome engineering is a powerful method for the study of bacterial virulence. With the availability of the complete genomic sequence of Bacillus anthracis, it is now possible to inactivate or delete selected genes of interest. However, many current methods for disrupting or deleting more than one gene require use of multiple antibiotic resistance determinants. In this report we used an approach that temporarily inserts an antibiotic resistance marker into a selected region of the genome and subsequently removes it, leaving the target region (a single gene or a larger genomic segment) permanently mutated. For this purpose, a spectinomycin resistance cassette flanked by bacteriophage P1 loxP sites oriented as direct repeats was inserted within a selected gene. After identification of strains having the spectinomycin cassette inserted by a double-crossover event, a thermo-sensitive plasmid expressing Cre recombinase was introduced at the permissive temperature. Cre recombinase action at the loxP sites excised the spectinomycin marker, leaving a single loxP site within the targeted gene or genomic segment. The Cre-expressing plasmid was then removed by growth at the restrictive temperature. The procedure could then be repeated to mutate additional genes. In this way, we sequentially mutated two pairs of genes: pepM and spo0A, and mcrB and mrr. Furthermore, loxP sites introduced at distant genes could be recombined by Cre recombinase to cause deletion of large intervening regions. In this way, we deleted the capBCAD region of the pXO2 plasmid and the entire 30 kb of chromosomal DNA between the mcrB and mrr genes, and in the latter case we found that the 32 intervening open reading frames were not essential to growth. PMID- 16369026 TI - A bacterial flagellin, Vibrio vulnificus FlaB, has a strong mucosal adjuvant activity to induce protective immunity. AB - Flagellin, the structural component of flagellar filament in various locomotive bacteria, is the ligand for Toll-like receptor 5 (TLR5) of host cells. TLR stimulation by various pathogen-associated molecular patterns leads to activation of innate and subsequent adaptive immune responses. Therefore, TLR ligands are considered attractive adjuvant candidates in vaccine development. In this study, we show the highly potent mucosal adjuvant activity of a Vibrio vulnificus major flagellin (FlaB). Using an intranasal immunization mouse model, we observed that coadministration of the flagellin with tetanus toxoid (TT) induced significantly enhanced TT-specific immunoglobulin A (IgA) responses in both mucosal and systemic compartments and IgG responses in the systemic compartment. The mice immunized with TT plus FlaB were completely protected from systemic challenge with a 200x minimum lethal dose of tetanus toxin. Radiolabeled FlaB administered into the nasal cavity readily reached the cervical lymph nodes and systemic circulation. FlaB bound directly to human TLR5 expressed on cultured epithelial cells and consequently induced NF-kappaB and interleukin-8 activation. Intranasally administered FlaB colocalized with CD11c as patches in putative dendritic cells and caused an increase in the number of TLR5-expressing cells in cervical lymph nodes. These results indicate that flagellin would serve as an efficacious mucosal adjuvant inducing protective immune responses through TLR5 activation. PMID- 16369028 TI - Differentially expressed and secreted major immunoreactive protein orthologs of Ehrlichia canis and E. chaffeensis elicit early antibody responses to epitopes on glycosylated tandem repeats. AB - Ehrlichia canis major immunoreactive proteins of 36 and 19 kDa elicit the earliest detectable antibody responses during the acute phase of canine monocytic ehrlichiosis. Genes encoding the major immunoreactive 36-kDa protein of E. canis and the corresponding ortholog of E. chaffeensis (47 kDa) were identified and the proteins characterized. The molecular masses of the strongly immunoreactive recombinant proteins were larger than predicted (26.7 and 32.9 kDa, respectively) but were consistent with those of the corresponding native proteins (36 and 47 kDa). Similar to other reported ehrlichial immunoreactive glycoproteins, carbohydrate was detected on the recombinant expressed proteins, indicating that they were glycoproteins. Both glycoproteins (gp36 and gp47) have carboxy-terminal serine/threonine-rich tandem repeat regions containing repeats that vary in number (4 to 16 repeats) and amino acid sequence among different isolates of each species. E. canis gp36 was recognized by early acute-phase antibodies (day 14), and species-specific antibody epitopes were mapped to C-terminal nonhomologous repeat units of gp36 and gp47. Periodate treatment of recombinant gp36 reduced the antibody reactivity, and nonglycosylated synthetic peptide repeat units from E. canis gp36 and E. chaffeensis gp47 were substantially less immunoreactive than corresponding recombinant peptides, demonstrating that glycans are important epitope determinants that are structurally conserved on the recombinant proteins expressed in Escherichia coli. E. canis gp36 and E. chaffeensis gp47 were differentially expressed only on the surface of dense-cored ehrlichiae and detected in the Ehrlichia-free supernatants, indicating that these proteins are released extracellularly during infection. PMID- 16369027 TI - Intercellular spreading of Porphyromonas gingivalis infection in primary gingival epithelial cells. AB - Porphyromonas gingivalis, an important periodontal pathogen, is an effective colonizer of oral tissues. The organism successfully invades, multiplies in, and survives for extended periods in primary gingival epithelial cells (GECs). It is unknown whether P. gingivalis resides in the cytoplasm of infected cells throughout the infection or can spread to adjacent cells over time. We developed a technique based on flow cytofluorometry and fluorescence microscopy to study propagation of the organism at different stages of infection of GECs. Results showed that P. gingivalis spreads cell to cell and that the amount of spreading increases gradually over time. There was a very low level of propagation of bacteria to uninfected cells early in the infection (3 h postinfection), but there were 20-fold and 45-fold increases in the propagation rate after 24 h and 48 h, respectively, of infection. Immunofluorescence microscopy of infected cells suggested that intercellular translocation of P. gingivalis may be mediated through actin-based membrane protrusions, bypassing the need for release of bacteria into extracellular medium. Consistent with these observations, cytochalasin D treatment of infected cells resulted in significant inhibition of bacterial spreading. This study shows for the first time that P. gingivalis disseminates from cell to cell without passing through the extracellular space. This mechanism of spreading may allow P. gingivalis to colonize oral tissues without exposure to the humoral immune response. PMID- 16369029 TI - Identification of the Vibrio vulnificus wbpP gene and evaluation of its role in virulence. AB - A wbpP gene encoding a putative UDP-N-acetyl-D-glucosamine C(4) epimerase was identified and cloned from Vibrio vulnificus. The functions of the wbpP gene, assessed by the construction of an isogenic mutant and by evaluating its phenotype changes, demonstrated that WbpP is essential in both the pathogenesis and the capsular polysaccharide biosynthesis of V. vulnificus. PMID- 16369030 TI - Inhibition of macrophage apoptosis by Neisseria meningitidis requires nitric oxide detoxification mechanisms. AB - Host-driven macrophage apoptosis contributes to innate immunity during bacterial infection. Neisseria meningitidis inhibits apoptosis in a variety of cells, but its impact on macrophage apoptosis is unknown. We demonstrate that N. meningitidis prevents macrophage apoptosis via genes encoding nitric oxide detoxification and a porin, PorB. PMID- 16369031 TI - Association between epithelial cell death and invasion by microspheres conjugated to Porphyromonas gingivalis vesicles with different types of fimbriae. AB - Microspheres (MS) conjugated to Porphyromonas gingivalis vesicles with type II fimbriae were the most efficient human epithelial cell invaders among the six types. Cell death was induced by MS, though becoming less frequent over time, with invasion efficiency partially related to cell death and gingipains likely the major cause. PMID- 16369032 TI - Contribution of sialic acid-binding adhesin to pathogenesis of experimental endocarditis caused by Streptococcus gordonii DL1. AB - An insertional mutation in hsa, the gene encoding the sialic acid-binding adhesin of Streptococcus gordonii DL1, resulted in a significant reduction of the infection rate of the organism and an inflammatory reaction in the rat aortic valve with experimental endocarditis, suggesting that the adhesin contributes to the infectivity of the organism for heart valves. PMID- 16369033 TI - The pap operon of avian pathogenic Escherichia coli strain O1:K1 is located on a novel pathogenicity island. AB - We have identified a 56-kb pathogenicity island (PAI) in avian pathogenic Escherichia coli strain O1:K1 (APEC-O1). This PAI, termed PAI I(APEC-O1), is integrated adjacent to the 3' end of the pheV tRNA gene. It carries putative virulence genes of APEC (pap operon), other E. coli genes (tia and ireA), and a 1.5-kb region unique to APEC-O1. The kps gene cluster required for the biosynthesis of polysialic acid capsule was mapped to a location immediately downstream of this PAI. PMID- 16369034 TI - Novel three-dimensional organoid model for evaluation of the interaction of uropathogenic Escherichia coli with terminally differentiated human urothelial cells. AB - Human bladder 5637 cells cultivated under microgravity conditions formed organoids that displayed characteristics of in vivo tissue-specific differentiation. Uropathogenic Escherichia coli (UPEC) strain CP9 colonized and penetrated the organoids and induced alpha-hemolysin-mediated exfoliation of uroepithelial cells. We propose these uro-organoids as models that simulate the interactions between UPEC and terminally differentiated human urothelium. PMID- 16369035 TI - Role of intimin-tir interactions and the tir-cytoskeleton coupling protein in the colonization of calves and lambs by Escherichia coli O157:H7. AB - Intimin facilitates intestinal colonization by enterohemorrhagic Escherichia coli O157:H7; however, the importance of intimin binding to its translocated receptor (Tir) as opposed to cellular coreceptors is unknown. The intimin-Tir interaction is needed for optimal actin assembly under adherent bacteria in vitro, a process which requires the Tir-cytoskeleton coupling protein (TccP/EspF(U)) in E. coli O157:H7. Here we report that E. coli O157:H7 tir mutants are at least as attenuated as isogenic eae mutants in calves and lambs, implying that the role of intimin in the colonization of reservoir hosts can be explained largely by its binding to Tir. Mutation of tccP uncoupled actin assembly from the intimin-Tir mediated adherence of E. coli O157:H7 in vitro but did not impair intestinal colonization in calves and lambs, implying that pedestal formation may not be necessary for persistence. However, an E. coli O157:H7 tccP mutant induced typical attaching and effacing lesions in a bovine ligated ileal loop model of infection, suggesting that TccP-independent mechanisms of actin assembly may operate in vivo. PMID- 16369036 TI - Listeria monocytogenes 10403S HtrA is necessary for resistance to cellular stress and virulence. AB - The HtrA serine protease has been shown to be essential for bacterial virulence and for survival after exposure to many types of environmental and cellular stresses. A Listeria monocytogenes 10403S htrA mutant was found to be sensitive to oxidative and puromycin-induced stress at high temperatures, showed a reduced ability to form biofilms, and was attenuated for virulence in mice. PMID- 16369037 TI - A sigma28-regulated nonflagella gene contributes to virulence of Campylobacter jejuni 81-176. AB - A Campylobacter jejuni 81-176 mutant in Cj0977 was fully motile but reduced >3 logs compared to the parent in invasion of intestinal epithelial cells in vitro. The mutant was also attenuated in a ferret diarrheal disease model. Expression of Cj0977 protein was dependent on a minimal flagella structure. PMID- 16369038 TI - Adoptive immunotherapy against experimental visceral leishmaniasis with CD8+ T cells requires the presence of cognate antigen. AB - CD8+ T cells have a protective role in experimental visceral leishmaniasis. However, the observation that inflammatory cytokines induce bystander activation of CD8+ T cells questions the need for antigen-dependent effector function. Here, we demonstrate that successful adoptive immunotherapy with CD8+ T cells is strictly dependent upon the presence of cognate antigen. PMID- 16369039 TI - Immunization with persistent attenuated Delta lpg2 Leishmania major parasites requires adjuvant to provide protective immunity in C57BL/6 mice. AB - Leishmania major parasites lacking the GDP-mannose transporter, termed Deltalpg2 parasites, fail to induce disease in mice but persist long-term. We previously found that Deltalpg2 organisms protect BALB/c mice from virulent L. major challenge. In contrast, we report here that Deltalpg2 parasites induce protective immunity in C57BL/6 mice only when administered with CpG-containing oligodeoxynucleotides, indicating that parasite persistence alone is not sufficient to maintain protective immunity to L. major. PMID- 16369040 TI - Molecular characterization of the Escherichia coli asymptomatic bacteriuria strain 83972: the taming of a pathogen. AB - Escherichia coli 83972 is a clinical asymptomatia bacteriuric isolate that is able to colonize the human urinary bladder without inducing an immune response. Here we demonstrate that one of the mechanisms by which this strain has become attenuated is through the mutation of its genes encoding type 1 and P fimbriae. PMID- 16369041 TI - A PE protein expressed by Mycobacterium avium is an effective T-cell immunogen. AB - Infection of mice with Mycobacterium avium or immunization with a novel PE gene expressed by M. avium (MaPE) showed that a dominant T-cell immune response was elicited. Immunization with an MaPE DNA vaccine protected mice against an aerosol challenge with Mycobacterium tuberculosis, suggesting that mycobacteria express PE antigens with cross-protective T-cell epitopes. PMID- 16369042 TI - New World monkey Aotus nancymae as a model for Campylobacter jejuni infection and immunity. AB - Three groups of six monkeys (Aotus nancymae) each were inoculated intragastrically with increasing doses of Campylobacter jejuni. Infection resulted in fecal colonization (100% of monkeys), dose-related diarrhea, and robust immune responses. Colonization duration and diarrhea rate were reduced upon secondary challenge. A. nancymae may be useful for studying anti Campylobacter vaccine efficacy. PMID- 16369043 TI - Correlation between lethal toxin-neutralizing antibody titers and protection from intranasal challenge with Bacillus anthracis Ames strain spores in mice after transcutaneous immunization with recombinant anthrax protective antigen. AB - Transcutaneous immunization of mice with recombinant protective antigen (rPA) of Bacillus anthracis resulted in significantly higher lethal toxin-neutralizing antibody titers than did intramuscular injection of alum-adsorbed rPA. Immunized mice were partially protected against intranasal challenge with 235,000 (10 50% lethal doses) Ames strain B. anthracis spores. A highly significant correlation was observed between toxin-neutralizing antibody titer and survival after challenge. Future experiments with rabbits and nonhuman primates should confirm the significance of protection by this vaccine strategy. PMID- 16369044 TI - Natural human antibody responses to Plasmodium vivax apical membrane antigen 1 under low transmission and unstable malaria conditions in Sri Lanka. AB - Plasmodium vivax apical membrane antigen 1, an important malaria vaccine candidate, was immunogenic during natural malaria infections in Sri Lanka, where low transmission and unstable malaria conditions prevail. Antibody prevalence increased with exposure in areas where malaria was or was not endemic. A marked isotype switch to cytophilic (immunoglobulin G1 [IgG1]/IgG3) antibodies was evident with increasing exposure exclusively in residents from areas of endemicity. PMID- 16369045 TI - Multiplexed profiling of candidate genes for CpG island methylation status using a flexible PCR/LDR/Universal Array assay. AB - DNA methylation in CpG islands is associated with transcriptional silencing. Accurate determination of cytosine methylation status in promoter CpG dinucleotides may provide diagnostic and prognostic value for human cancers. We have developed a quantitative PCR/LDR/Universal Array assay that allows parallel evaluation of methylation status of 75 CpG dinucleotides in the promoter regions of 15 tumor suppressor genes (CDKN2B, CDKN2A, CDKN2D, CDKN1A, CDKN1B, TP53, BRCA1, TIMP3, APC, RASSF1, CDH1, MGMT, DAPK1, GSTP1, and RARB). When compared with an independent pyrosequencing method at a single promoter, the two approaches gave good correlation. In a study using 15 promoter regions and seven blinded tumor cell lines, our technology was capable of distinguishing methylation profiles that identified cancer cell lines derived from the same origins. Preliminary studies using 96 colorectal tumor samples and 73 matched normal tissues indicated CpG methylation is a gene-specific and nonrandom event in colon cancer. This new approach is suitable for clinical applications where sample quantity and purity can be limiting factors. PMID- 16369047 TI - Regulation of voltage-gated Ca2+ channels by calmodulin. AB - Calmodulin, a highly versatile and ubiquitously expressed Ca2+ sensor, regulates the function of many enzymes and ion channels. Both Ca2+-dependent inactivation and Ca2+-dependent facilitation of the voltage-gated Ca2+ channels Cav1.2 and Cav2.1 are regulated through an interaction with Ca2+-bound calmodulin. This review addresses the functional regulation of Cav1.2 and Cav2.1 by calmodulin and discusses how Ca2+ binding to a single calmodulin molecule can regulate opposing functions of the voltage-gated Ca2+ channels. PMID- 16369046 TI - The prion protein gene in humans revisited: lessons from a worldwide resequencing study. AB - Ample evidence has accumulated showing that different coding variants of the PRNP gene confer differential susceptibility for prion diseases. Here we evaluate the patterns of nucleotide variation in PRNP exon 2, which includes all the protein coding sequence, by resequencing a worldwide sample of 174 humans for 2378 bp. In line with previous studies, we found two main haplotypes differentiated by nonsynonymous substitution in codon 129. Our analyses reveal the worldwide pattern of variation at the PRNP gene to be inconsistent with neutral expectations, indicating instead an excess of low-frequency variants, a footprint of the action of either positive or purifying selection. A comparison of neutrality test statistics for PRNP with other human genes indicates that the signal of positive selection on PRNP is stronger than expected from a possible confounding genome-wide background signal of population expansion. Two main conclusions arise from our analysis. First, the existence of an ancient, stable, balanced polymorphism that has been claimed in a previous study and related to cannibalism can be rejected and is shown to be due to ascertainment bias. Second, our results are consistent with a complex history of selection including mainly positive selection, even if short local periods of balancing selection (Kuru-like episodes), or even a weak purifying selection model, are consistent with our data. PMID- 16369048 TI - Multiplex analysis of intracellular signaling pathways in lymphoid cells by microbead suspension arrays. AB - Phosphorylation analysis of signaling proteins is key for examining intracellular signaling pathways. Conventional biochemical approaches, e.g. immunoprecipitation, Western blot, and ELISA, have played a major role in elucidation of individual signaling events. However, these methods are laborious, time-consuming, and difficult to adapt for high throughput analysis. A multiplex approach to measure phosphorylation state of multiple signaling proteins simultaneously would significantly enhance the efficiency and scope of signaling pathway analysis for mechanistic studies and clinical application. This report describes a novel multiplex microbead suspension array approach to examine phosphoproteomic profiles in lymphoid cells. In the Jurkat T-cell leukemia line, the multiplex assay enabled targeted investigation of phosphorylation kinetics of signal transduction from receptor proximal events (tyrosine phosphoproteins CD3, Lck, Zap-70, and linker for T-cell activation) to cytosolic events (serine/threonine phosphoproteins Erk and Akt) to transcription factors (serine/threonine phosphorylated Rsk, cyclic AMP-response element-binding protein, and STAT3). To broaden the application of the multiplex analysis, signaling pathways were also studied in B-cell lymphoid tumor lines that included chronic lymphocytic leukemia lines. In these cell lines, multiplex suspension array enabled phosphoproteomic analysis of signaling cascade mediated by Syk, a homolog of Zap-70. Results obtained by multiplex analysis were confirmed by immunoprecipitation and Western blot methods. The examples of T-cell and B-cell signaling pathway analyses in this report demonstrate the utility of the multiplex suspension arrays to investigate phosphorylation dynamics and kinetics of several signaling proteins simultaneously in signal transduction pathways. PMID- 16369049 TI - Synthesis and biological properties of the fluorescent ether lipid precursor 1-O [9'-(1''-pyrenyl)]nonyl-sn-glycerol. AB - The synthesis of an omega-pyrene-labeled 1-O-alkyl-sn-glycerol was performed using a chirospecific method starting from R-(-)-2,3-O-isopropylidene-sn glycerol. The product, 1-O-[9'-(1''-pyrenyl)]nonyl-sn-glycerol (pAG), is a fluorescent ether lipid that has a pyrene moiety covalently attached at the alkyl chain terminus. pAG was taken into CHO-K1 cells and a plasmalogen-deficient variant of CHO-K1, NRel-4. This variant is defective in dihydroxyacetonephosphate acyltransferase, which catalyzes the first step in plasmenylethanolamine (PlsEtn) biosynthesis. pAG was incorporated primarily into ethanolamine and choline phospholipids as well as a neutral lipid fraction tentatively identified as alkyldiacylglycerol. NRel-4 accumulated more fluorescence in the phospholipid fraction than CHO-K1, specifically in the ethanolamine phospholipids. Analysis of the fluorescent lipids showed that 93% of the pAG was incorporated into glycerolipids with the ether bond intact. Although the addition of 20 microM 1-O hexadecyl-sn-glycerol to the medium fully restored PlsEtn biosynthesis in NRel-4 cells, pAG only partially restored PlsEtn synthesis. Incubation of cells with pAG followed by irradiation with long-wavelength (>300 nm) ultraviolet light resulted in cytotoxicity. NRel-4 cells displayed an increased sensitivity to this treatment compared with CHO-K1 cells. This photodynamic cytotoxicity approach could be used to select for mutants that are defective in downstream steps in ether lipid biosynthesis. PMID- 16369050 TI - Substrate specificity of lysophosphatidic acid acyltransferase beta -- evidence from membrane and whole cell assays. AB - Membranes of mammalian cells contain lysophosphatidic acid acyltransferase (LPAAT) activities that catalyze the acylation of sn-1-acyl lysophosphatidic acid (lysoPA) to form phosphatidic acid. As the biological roles and biochemical properties of the six known LPAAT isoforms have yet to be fully elucidated, we have characterized human LPAAT-beta activity using two different assays. In a membrane-based assay, LPAAT-beta used lysoPA and lysophosphatidylmethanol (lysoPM) but not other lysophosphoglycerides as an acyl acceptor, and it preferentially transferred 18:1, 18:0, and 16:0 acyl groups over 12:0, 14:0, 20:0, and 20:4 acyl groups. The fact that lysoPM could traverse cell membranes permitted additional characterization of LPAAT-beta activity in cells: PC-3 and DU145 cells converted exogenously added lysoPM and (14)C-labeled 18:1 into (14)C labeled phosphatidylmethanol (PM). The rate of PM formation was higher in cells that overexpressed LPAAT-beta and was inhibited by the LPAAT-beta inhibitor CT 32501. In contrast, if lysoPM and (14)C-labeled 20:4 were added to PC-3 or DU145 cells, (14)C-labeled PM was also formed, but the rate was neither higher in cells that overexpressed LPAAT-beta nor inhibited by CT-32501. We propose that LPAAT beta catalyzes the intracellular transfer of 18:1, 18:0, and 16:0 acyl groups but not 20:4 groups to lysoPA. PMID- 16369052 TI - [Minimally invasive surgery in treatment of pancreas pseudocysts]. AB - Results of treatment of 68 patients with cystic formations of a pancreas are presented. Minimally invasive methods -- US-guided transcutaneous puncture (d < 5 cm; I group) and drainage (d > 5 cm; II group) and use of Ivshin's tools has allowed to achieve positive results. For small intrapancreatic pseudocysts (d < 5 cm), it was applied transcutaneous puncture with replacement of contents by protease inhibitors. The obtained material was subjected to cytological, biochemical and bacteriological examination. High level of diastase indirectly indicated to the presence of connection with the main pancreatic duct. For cysts of large sizes (d > 5 cm), transcutaneous drainage with subsequent cystography for definition of the possible connection of a cyst cavity with a pancreatic duct was carried out. When the communication with the main pancreatic duct was absent, drainage of the cavity was carried out. Presence of connection of formations with the main pancreatic duct was the indication of US and endoscope-guided introduction drainage in the cystic formation through a stomach -- transgastric drainage. The mean follow-up period ranged from 6 months till 2.5 years. Excellent results were obtained in 69.2% cases and good -- in 30.8% in the I group of patients. In the II group there were received excellent results in 51.7% cases, good results -- in 41.4% and satisfactory -- in 6.9. The level of quality of life approached to maximal index in both groups. Patients promptly came back to habitual social and intellectual activity. PMID- 16369051 TI - Mechanism-based inactivation and reversibility: is there a new trend in the inactivation of cytochrome p450 enzymes? AB - Recent studies with cytochrome P450 (P450) enzymes from the 2E and 2B subfamilies have shed light on what may be a new trend in the mechanism-based inactivation of P450s: reversibility. The reversible inactivation of P450-type enzymes was first reported in the mid-1990s by Dexter and Hager [Dexter AF and Hager LP (1995) J Am Chem Soc 117:817-818], who studied the transient heme N-alkylation of chloroperoxidase by allylbenzene and 1-hexyne. While characterizing small tert butyl acetylenes as mechanism-based inactivators of P450s 2E1 and 2B4, Hollenberg and coworkers observed the reversible inactivation of an acetylene-inactivated T303A mutant of P450 2E1. The mechanism of reversibility was a combined product of the structure of the inactivator and the positioning of conserved amino acid residues, threonine 303 (alanine in the mutant) and glutamate 302, in the enzyme active site. Reversibility was also observed with both wild-type P450 2B4 and the T302A mutant of 2B4, although this inactivation and reversibility did not seem to depend on the T302 residue. Subsequent studies have attempted to elucidate the chemical/structural requirements of the inactivator in determining reversibility and have shown that both the size and the chemical nature of functional groups play an important role. At this time, reversibility has only been observed with P450 2E and 2B enzymes during their mechanism-based inactivation by terminal alkynes. Future studies with P450s from other subfamilies and structurally distinct inactivators will greatly aid our understanding of the molecular and chemical determinants of reversibility. PMID- 16369053 TI - [Container method in hair transplant megasessions]. AB - Presently, it is imperative to obtain a maximum survival rate of follicular units during large-scale transplantation, as the result of hair transplantation depends not only on the quantity of implanted grafts but on the number of survival rate of follicular units in-vitro in continuous moisture during the extended process of hair transplantation. To provide this, prepared grafts are kept in a cold saline solution. The only technological stage of hair transplantation during which grafts exist without a preserving solution is the immediate process of implantation. At this time, grafts placed on the surgeon's gloves are subjected to the harmful drying influences. For exclusion of grafts dehydratation during implantation, minimization crashing injury and shortening of the operation time we have recommended simple adaptation of a so-called Container Method. The goal of this work was the introduction of Container Method and estimation of it's effectiveness in large hair transplantation sessions. Container Method allows reducing grafts dehydratation, iatrogenic injury and warming during implantation. According to clinical observations using above mentioned method gives reduction of operation time by 41%. That fact that containers are very simple adaptations and don't require staff's special training gives a good opportunity to use them widely in the future in hair transplant megassesions. PMID- 16369054 TI - [Risk of iugr syndrome development during preeclampsia of the pregnant]. AB - We have conducted a case-control retrospective study to determine the risk of Intra Uterine Growth Retardation (IUGR) in pregnant women with preeclampsia. We have found that IUGR is one of the most prominent features of preeclampsia and occurs in 27% of cases (Odds Ratio 1.20). IUGR occurs during fetal-placental insufficiency (ultrasound structural changes of placenta such as aging and calcifications take place in 67.8% of cases, oligohydramnia in 42.8% of cases, changes of hemodynamics in maternal -- fetal placental unit -- in 82.3% of cases; deviation from the norm of resistance index and pulse index was 1.5-2 times higher). IUGR was accompanied with the poor functional state of the fetus (85.7% according to the cardiotocographic monitoring); morphological investigation of placenta showed dystrophic and destructive changes, which finally leads to the high rate of morbidity during neonatal period (hypoxic-ischemic encephalopathy -- 66%, respiratory distress syndrome -- 36%, pneumonia and atelectasis --14%) and fetal functional immaturity confirmed by EEG. PMID- 16369055 TI - The expression of cadherin e and clinical prognostic factors in uterine endometrioid adenocarcinoma. AB - The aim of our study was to evaluate the association between the expression of E cadherin and clinical prognostic factors in uterine endometrioid adenocarcinoma of different histological grade. We have studied 104 postmenopausal women with diagnosis of endometrioid adenocarcinoma. We evaluated the presence of obesity and vaginal bleeding. Surgical specimens were fixed in 10% neutral buffered formalin solution and embedded in paraffin. 4 mm sections were stained by hematoxylin-eosin, von Gieson, and histological type of cancer, metastatic involvement of lymph nodes and depth of myometrial invasion were evaluated. Histological grade of cancer was assessed by FIGO grading system. All samples were analyzed by immunohistochemistry for E-cadherin (Dakocytomation). We assessed the number of E-cadherin-positive and negative tumor cells and degree of positivity (low, moderate, high). Histological study by hematoxylin-eosin has showed grade 1 endometrioid carcinoma in 35 cases (33.7%, group I), grade 2 adenocarcinoma in 44 cases (42.3%, group II), and grade 3 adenocarcinoma in 25 cases (24%, group III). Our results suggest that the loss of E-cadherin expression is associated with a higher histological grade of uterine endometrioid adenocarcinoma, depth of myometrial invasion, lymph node positivity, coexistence of obesity and vaginal bleeding. It seems that local invasion and metastatic spread of tumor should be preceded by the loss of E-cadherin expression in tumor cells, which progressively occurs in carcinogenesis. Therefore, E-cadherin negativity can be used as a poor prognostic factor and more aggressive chemotherapy regimen should be used. PMID- 16369056 TI - [Combined usage of duphaston and reaferon for infertility treatment in patients with endometriosis]. AB - A new method of treatment of endometriosis includes combination of surgical and hormonal therapy. Surgical method included excision of tissue with endometriosis, while the hormonal therapy involved the combination of dufaston and reaferon-EC (recombinant interferon alpha 2b). The aim of this study was to develop effective method of treatment of infertility caused by endometriosis using combination of dufaston and reaferon. There were 114 patients under observation who suffered from endometriosis and infertility. At the first stage of treatment thermokouterisation of area with endometrosis has been used. At the second stage of treatment all patients were divided into different clinical groups. To 34 patients from the study group reaferom-EC has been administered on the 5th, 8th, 10th, 12th, 14th days of menstrual cycle. The control group was represented by 80 patients, which was divided into two subgroups, 40 patients in each group. The patients from the first subgroup got hormonal therapy 20 mg dufaston from the 5th to 25th days of menstrual cycle, while the patients from the second subgroup got 20 mg dufaston from the 15th-20th to 25th days. The treatment was successful among the patients who got the combination of reaferon and dufaston, the frequency of pregnancy in the study group reached 52% vs. 27.3% in the control group (p>0.05). Those patients who had so called "small form" of endometriosis became pregnant in 34% cases in the study group and in 22.7% cases in the control group. Patients who had endometriosal cystoma became pregnant in 18.7% in the study group, but only in 9.4% in the test group, which shows that the treatment is less effective among these categories of patients. As a result of our study it is proved that the method of combination of reaferon and dufaston gives a chance to increase therapeutic effect and the number of pregnancy among the patients with endometriosis. PMID- 16369057 TI - [Diagnostic principles of gonadal dysgenesis in adolescents]. AB - 103 patients aged 11-20 with short stature and/or with delayed puberty were studied. Among all patients classical form of gonadal dysgenesis was diagnosed in 31 patients (karyotype 45,X- 17 patients, 45,X/46,XX- 14), "mixed" gonadal dysgenesis in one patient (karyotype 45,X/46,XY)-, "pure" gonadal dysgenesis- in 5 cases (karyotype 46,XY-in 4 , 45,X -- in one). The majority of cases (66) were diagnosed as ovarian dysgenesis with the mosaic karyotype- 46,XX/45,X. In the clinical diagnostics of ovarian dysgenesis some difficulties were aroused because of slight reduction of height and delayed sexual development, mild appearance absence or somatic anomalies. The main diagnostic criteria for the various forms of gonadal dysgenesis are detection of characteristic karyotype and hypogonadotropic hypogonadism. Characteristic phenotype is more informative in the cases of classical and "mixed" forms of gonadal dysgenesis. Early diagnosis of gonadal dysgenesis is very important for effective correction of height and sexual development. Timely begin hormonal therapy may avoid complications induced bihypoestrogenia, such are: osteoporosis, cardiovascular diseases and etc. Karyotype investigation and in the cases of revealing Y chromosome material, and therefore gonadectomy must precede the beginning of hormonal therapy. PMID- 16369058 TI - [Influence of connective tissue dysplasia on the clinical course of chronic catarrhal gingivitis]. AB - "MASS"- phenotype or dysplasia of the connective tissue has been studied for many years, but it still remains the actual problem in clinical and theoretical medicine. Expansive integration of clinical genetics and molecular biology into the medicine has revealed the most complicated and multistage structure of the pathogenesis of this disease. The aim of our study was focused on the investigation of "MASS" phenotype and its influence on the stomatological disease such as the early stage of periodontitis -- chronic catarrhal gingivitis. Two groups of subjects with chronic catarrhal gingivitis were examined. Subjects in the first group had the chronic catarrhal gingivitis but did not belong to "MASS" phenotype, while the subjects in the second group had the chronic catarrhal gingivitis and belonged to "MASS" phenotype at the same time. Comparing clinical characteristics of these two groups, the clinical course of catarrhal gingivitis among subjects with "MASS" phenotype was characterised by more acute dynamics, weaker response to conventional treatment and more frequent recidives. PMID- 16369059 TI - [Microbiological investigations and studies of phagocytic activities of peripheral neutrophils during the treatment of parodontitis by Unimag]. AB - During the treatment by Unimag (UN), quantity of microbes in the mouth cavity of patients with periodontitis (PD), significantly decreases in comparison with the patients treated by traditional scheme. That is due to direct and indirect influence of UN on the pathogenic microorganisms. During the treatment of patients with PD by UN, quantity of Gram-negative microbes gradually decreases and their substitution by Gram-positive microbes, typical for mouth cavity, takes place. On the background of the treatment by UN, phagocytic activity (PA) of polynuclear cells (PC) increases. In comparison with the patients treated by traditional scheme, increases both phagocytic number and number of active neutrophils. On 14-15 days after beginning of treatment of patients with PD by traditional scheme, PA of PC does not change significantly. Reduction of the microbial number in the mouth cavity and the active substitution of Gram-negative microbes by Gram-positive microorganisms during the treatment of patients with PD by UN, have prognostic importance and together with the reinforcement of PA of PC indicate to the improvement of the therapeutic effect and shortening of the duration of the treatment. PMID- 16369060 TI - [Predictor of early thrombolysis in acute coronary syndrome by estimation of the QT interval variability on 12 lead standard electrocardiogram]. AB - It has been stated that decreased dispersion of QT interval (QTd) on 12 lead ECG is associated with successful thrombolysis (TH) in acute myocardial infarction but it is not clear whether QTd can predict early successful TH. The goal of the investigation was to evaluate significance of QTd and QT interval variability between adjacent precordial leads (QTdl) for prediction of successful early TH in acute coronary syndrome (ACS). Two groups of patients after 6 hours from the beginning of acute coronary attack with elevation of ST segment on ECG were under investigation: I group -- 26 patients with resolution of ST> or =70% after 60-90 min from the starting of TH and II group -- 21 patients with no resolution of ST. QTd was defined as the difference between maximal and minimal QT interval and QTd as the difference between adjacent precordial ECG leads. QTd is a marker of successful TH but does not predict its efficiency, QTde<50 predicted TH with 80% sensitivity, 85% specificity and 87% positive predictive value. PMID- 16369061 TI - New type of pulsate flow system for artificial heart-lung bypass. AB - In the artificial system of heart-lung bypass, used during the surgery on stopped heart, a com-mon problem-causing device is a roller pump. Inexpensive, simple, yet dependable device is in the process of development, with characteristics maximally approximated to the physiologic parameters of the organism's natural heart. Substituting for the roller pumps, this device consists of two reservoirs. Hermetic reservoirs are interconnected in parallel. They also connect to other parts of the system with blood tubing. A pneumo console for artificial ventricles represents the control system and ensures pulsate blood flow in the bio-models. The developed device provided not only an opportunity to assign frequency of a pulsation, but also an opportunity of control in each output volume. For fulfillment of maximum physiologic blood flow, it is necessary to provide cardio synchronized mode of perfusion. PMID- 16369062 TI - [Fluctuation of C-reactive protein levels in acute coronary syndrome]. AB - The aim of the study was to find out whether early use of atorvastatin in patients with acute coronary syndrome is associated with rapid changes of plasma levels of the marker of inflammation -- C-reactive protein. 32 patients (15 male, 17 female; no more then 3 hours from pain onset, age 61+/-10 years) treated with AHA recommendations, were assigned to the two groups. In the I group (n=17) patients received atorvastatin (liprimar) 40 mg/day, II group (n=15) represented the control group. Plasma level of C-reactive protein (hs-CRP) was assessed by highly sensitive method at baseline, on days 7 and 14. Baseline clinical characteristics and C-reactive protein level were similar in all groups. C reactive protein level was increased in both groups from the baseline on day 7 (p<0.01). In group I C-reactive protein level was decreased by 16.54% from the baseline on day 14 (from 22.48+/-1.65 to 18.76+/-1.55 mg/l, p=1.7). In II group C reactive protein level was increased by 9.12% from baseline on day 14 (from 22.79+/-1.67 to 24.87+/-1.24 mg/l, p<0.0005). In patients with acute coronary syndrome early use of atorvastatin was associated with rapid decrease of C reactive protein level. PMID- 16369063 TI - Flow-mediated dilation in patients with left ventricular diastolic dysfunction. AB - The aim of the study was to evaluate the endothelium dependent FMD of the brachial artery in patients with LVDD. Endothelial function was assessed in 36 male patients (mean age 49+/-6 years) with LVDD due to CAD (22 patients) and essential hypertension (14 patients) as well as in 18 male patients (control group) without LVDD (12 patients with CAD and 6 with hypertension) of matched age. Our studies had showed significant decrease of FMD in patients with LVDD compared to those without LVDD (4.67%+/-1.68 vs. 6.12%+/-2.54, p<0.05). At the same time FMD in both groups appeared to be significantly lower than in healthy subjects. Post-ISDN vasodilation was almost similar in both groups of patients and lower than in controls. The post-ISDN vasodilatation did not differ regarding presence of LVDD or causative disease (CAD or arterial hypertension) and its extent was lower than in healthy controls. A weak correlation was found between FMD and post-ISDN vasodilatation (r=0.231, p>0.05). Data obtained suggest that endothelial dysfunction exists in patients with diastolic dysfunction. Based on these results it is considered that patients with early stages CAD and arterial hypertension depressed FMD might be indicator of LVDD. PMID- 16369064 TI - [Role of boundary neuro-psychical pathologies in the development of disruptive behavior in adolescents]. AB - Behavioural disorders in youths represent a very serious social problem and create discomfort both for the society and for adolescents themselves. This problem became very actual in adolescents because of identity crisis incident in this age. Boundary neuro-psychical pathologies seem to play a serious role as a cause of forming deviations of behaviour. Burden of these pathologies is rather high in both of groups examined by us (delinquent adolescent and adolescent with school disadaptation ) -- 49-50%, but structure and severity of these pathologies are very different: in the population of delinquent adolescents there are more severe pathologies such as residual cerebral-organic dysfunction, signs of personal anomaly with symptoms of excitability, retardation of psychical evolution with marked cognitive deficiency, whereas in adolescent with school disadaptation prevail personality disorders, phenomena of pathological puberty crisis and retardation of psycho-social evolution. PMID- 16369065 TI - [Clinical and experimental-psychological features of depression in late onset schizophrenia patients with Kandinski Klerambo's syndrome]. AB - Clinical-psychopathological and experimental-psychological investigation of depressed late onset schizophrenia patients with Kandinski Klerambo's syndrome made possible to determine the most frequent subgroups of depressive states: a) apatho-adynamic depression, b) depression with cenestopathy and hypochondriac symptoms, c) depression with depersonalization and de-realization, d) melancholic depression. The experimental psychological examination with method of fixed set by D.N Uznadze revealed relatively intact personality features that could be explained by different aging factors. Analyzing correlation between fixed set characteristics among the depressed schizophrenia patients with passivity phenomena we could assume, that basically the fixed set among those patients is stable, persistent, static, steady, constant, generalized and in some cases, in the presence of dis-automation of sensory integrity, it may be local. PMID- 16369066 TI - Effect of magnesium and furosemide on bronchial asthma. AB - In recent years along with the conventional bronchodilatators the data of so called "extraordinary bronchodilatators" have appeared, among which there are furosemide and magnesium sulphate. However these data concern their bronchoprotection properties (those often are contradictory), and about bronchodilatative properties of these medications nothing is known. Proceeding from above stated the purpose of our study was the investigation of bronchodilative effectiveness of magnesium and furosemide delivered through an inhalation route in treatment of mild and severe course of bronchial asthma. The study was performed on patients with bronchial asthma in age of 20-70 (39 persons), who were in-patients of Allergology and Clinical Immunology Center of TMSU. For the study the double-blind placebo-controlled trial with crossover design and parallel study methods were used. As primary variables following data were assumed: the forced expiratory rate, respiratory rate and heart rate. The analysis of obtained results by t-statistics methods has shown that the effectiveness of magnesium comparatively to placebo has been insignificant in quantitative improvement (though statistically reliable), and the furosemide has comparatively better bronchodilative effect (particularly in case of PEF). PMID- 16369067 TI - [Functional status of haemostasis system in patients with visceral leishmaniasis]. AB - During the last decades in Georgia was observed significant increase of cases of visceral leishmaniasis and fight against this disease became important problem as far as the management of this disease is rather problematic. According to references and our clinical experience patients with visceral leishmaniasis are predisposed to bleeding. The objective of our study was the assessment of functional status of hemostasis related to the degree of clinical severity. We have studied platelet count, activated partial thromboplastin time (APTT), prothrombin time, thrombin time, plasma concentration of fibrinogen, the soluble fibrin-monomeric complexes (SFMC), fibrinogen/fibrin degradation products (D dimer) and anticoagulant protein C. Haemostatic functional tests were studied in 45 patients with visceral leishmaniasis before and after treatment (with 20-25 day intervals). Before treatment the reduction of platelet count was observed in 95%. Prolonged APTT and prothrombin time was found in severe forms of the disease. Thrombin time prolonged in 45.7%, SFMC level was increased in 80% (p=0.003) and D-dimer level in 95.6% (p=0.023). Protein C was in normal value in 73%. The results indicate that leishmania infection affects primary haemostasis, coagulation and fibrinolysis and these alterations are related to the severity of clinical symptoms. Investigation of SFMC and D-dimer showed that in case of visceral leishmaniasis activation of intravascular coagulation takes place, particularly during the severe forms of the disease, study of these markers is of the diagnostic and prognostic importance and the treatment at an early stage of infection may potentially avoid the possibility of developing an uncompensated DIC. PMID- 16369068 TI - [Peculiarities of epileptic syndrome in children with metabolic disorders of nervous system]. AB - Metabolic diseases of the nervous system vary considerably in their clinical and pathological aspects. In neurological presentations of these disorders dominate mental retardation and epileptic syndrome. We have studied 27 patients of age from 3 months to 3 years: PKU -- 15 cases; homocystinuria -- 4; hyper-prolinemia - 1; methylmalonic acidemia -- 5 and combined disorders -- 2. Epileptic syndrome was revealed in 21 patients, mental retardation in 1, spasticity in 5 and ataxia in 1 patient. Epileptic syndrome was presented with generalized seizures (grand mal -- 6 cases, myoclonic absences -- 13 cases) and partial seizures (simple motor -- 2 cases). Investigations did not found reliable correlations between certain forms of enzymophaties and EEG patterns. Patients were treated by pathogenic (dietary management with protein-modified diet and vitamin therapy) and symptomatic (anticonvulsants) treatment. We have achieved the positive therapeutic effect by pathogenic and anticonvulsive treatment in 11 patients. All these patients were from the first group (1-3 year). The best outcome was observed in the cases of the early diagnosed PKU. The most severe mental retardation and resistant epilepsy were revealed in patients with combined disorders of metabolism and vitamin-non-responsive forms of MMA and HCS. PMID- 16369069 TI - [Comparative analysis of prognostic value of epileptic encephalopathies and symptomatic epilepsy in children with cerebral palsy]. AB - Cerebral dysfunction due to epilepsy is not currently well-understood and needs to be further investigated. We have observed patients with static encephalopathy (cerebral palsy) of nearly equal severity with epileptic encephalopathy and symptomatic epilepsy and have had carried out comparative analysis of prognosis among these groups. The work is based on the results of investigation of 121 patients admitted to the Center of child neurology and neurorehabilitation during the period 2000-2005. 69 patients with epileptic encephalopathies were included into the study groups according to following criteria: I group -- cases with early starting seizures (before 1 year of life) and II group -- starting seizures after 1 year of life. The control group involved 52 patients with associated cerebral palsy of different severity and symptomatic epilepsy. The study and control groups were compared prognostically. According to our data, prognosis and outcome of epilepsy was closely associated with the time of onset of epilepsy syndromes: early started (within the first year) epileptic encephalopathies display the highest correlation with poor prognosis; the risk of development of epileptic encephalopathies is the highest in patient with cerebral palsy and early starting of seizures. PMID- 16369070 TI - [Youth health behavior survey]. AB - High-risk behaviours -- smoking, drug use, alcohol use and etc. are the main risk factors of non-communicable diseases and main reasons of morbidity and mortality in many countries of the world. We conducted the "Youth Health Behaviour Survey" among the students in Tbilisi, Mtskheta and Dusheti. The survey included questioning of students (14-17 years old) by modified questionnaire. There were questioned 300 students, 100 in each region. Mtskheta is very close to Tbilisi, it's a peri-urban area; Dusheti is quite far rural area. The survey results show that teenagers both in central and peripheral areas are becoming familiar with alcohol in early ages and using alcohol is several times higher in Tbilisi. In Tbilisi as in Mtskheta and Dusheti students use wine and spirits with same frequency. Smoking is a bit high in Tbilisi and Mtskheta; regular smoking starts at age 13-14, boys are smoking more than girls. Drug use wasn't identified. Sexual intercourse had almost half of boys from Tbilisi and about one third of boys in Mtskheta and Dusheti. Safe sex methods more frequently are used in Tbilisi. Propaganda of healthy life style is essential in supporting healthy youth development. It's very important to set different interventions in different regions of Georgia, to adapt materials to real situation. PMID- 16369071 TI - [Major pathogenic links of atherosclerosis]. AB - The experimental and clinical data concerning pathogenesis of the atherosclerosis are summarized and analyzed in this article. Major concepts that explain initiation and progressive growth of atherosclerosis such as lipid infiltrations, response to disturbing factors, "response on the keeping of particles" and inflammatory processes are discussed. These concepts are considered as base for integral theory of atherosclerosis according which the inflammatory process in atherosclerosis are the result of the universal response reaction of endothelium to the various disturbing risk factors. Chronic inflammation leads to complex cellular and molecular interactions among cells derived from the endothelium, smooth muscle and several blood cell components and causes oxidative stress, proliferation of smooth muscle cells, oxidative modification of LDL, uptake and macrophage foam cell formation, endothelium dysfunction. Major pathogenic links of atherosclerosis, such as inflammation, oxidative stress, oxidative modification of LDL, lipid infiltration, endothelial dysfunction closely interact, forming close vicious circles which leads to metabolic and morphological disturbances, re-modulation of blood vessels, cardiovascular diseases and such complication as cardiac infarction and stroke. Pathogenic peculiarities of atherosclerosis are the theoretic base to the elaboration of therapeutic strategy. Endothelium may be discussed as a new therapeutic target in atherosclerosis. So far as the leukotrienes play an important role in inflammatory processes, it is suggested that the leukotrienes may be as a potential therapeutic target in cardiovascular diseases. PMID- 16369072 TI - Some opinions about matter and material substances: from inanimate system -- to living according to A. Einstein general theory of relativity. AB - During transformation from inanimate to living, change of the space position of the matter causes the change of the field, as the space does not exist without the field, therefore the time-space as the properties of material substances, should undergo certain changes. The outside inanimate system, in this case a matrix, has its own time. The living system, in this case a cell, where the matter undergoes space conformation with the change of field and space-time, has its own time and it has begun to flow more rapidly than in matrix. From the surface of the body, from different energetic reservoirs oppositely charged matter substances following from special transport systems from the life system transmitted into lifeless one and change their matter space conformation, create transmission gradient that is the gradient border of time from lifeless system into live. In the case of a human, hypothetically, the gradient system of time must be of a complex scheme counting the inter-transformation and interaction gradients of outer and inner abdominal systems. Subconscious and consciousness by means of special links and messages, information selection interact and form unique connection between the systems. Subconscious serves for accelerated time system. Conscious by means of permanent contact with the environment collects and reacts in matrix time system By interconnection of these two systems ideal adaptation with the environment takes place. Time difference gradient system is an additional energy factor, by means of which respective ordered geometrical structures special for the given types are formed. The living organism is an inter-regulated interconnection global system resulting from the changes of matter and material substances space configuration. PMID- 16369073 TI - [Plaferon LB as a modulator of the activity of transport protein complexes of mitochondrial respiratory chain]. AB - Mitochondrial energogenesis plays key role in the vital activity of cells. Regulation of its intensity is especially important for the maintenance of function of tissues and body internal organs under different pathological conditions. There are numerous studies demonstrating modulator effect of domestic preparation, plaferon LB (USA patent number WO02/ 12444 A2) on the intensity of mitochondrial respiration in vivo at different pathological processes. In order to determine the mechanism of modulator effect of plaferon LB we have studied effectiveness of its effect on the intensity of electron transport in mitochondrial respiratory chain, inhibited by different toxins, in isolated myocardial sections in vitro. Isolated sections (100mg) of myocardium of white rats were placed in the buffer solution with glutamate and DNF. Obtained dredge was distributed in equal portions to the different cells, where amital -- inhibitor of I complex of mitochondrial electron transport chain or potassium cyanide, inhibitor of IV complex of mitochondrial electron transport chain, or plaferon LB have been added in different consecution with 2 min intervals (USA patent number WO02/ 12444 A2). We discovered that plaferon LB rehabilitates electron transport on the sites of I and IV complexes of mitochondrial chain that leads to a decrease in free radical production and normalization of cell redox status. This is the mechanism through which the preparation regulates the intensity of mitochondrial respiration and tissues energogenesis. PMID- 16369074 TI - The age related changes in cell cycle of mice cardiomyocytes. AB - Although rapid progress is being made in many areas of molecular cardiology, issues pertaining to the origins of heart-forming cells, the mechanisms responsible for cardiogenic induction, and the pathways that regulate cardiomyocyte proliferation during embryonic and adult life remain unanswered. The present study was performed to investigate the age related changes in cardiomyocytes' cell cycle in white mice. 18 white mice were used. The animals were distributed in three age groups: juveniles (2 months old), adults (10 months old) and senescents (18 months old). The animals were killed under ether narcosis. The heart was removed. Number of diploid, tetraploid, polyploid (octaploid, 16 n and 32 n), apoptotic nuclei and nuclei in S phase were counted by a Becton Dickinson (Mountain View, CA) FACScan flow cytometer. Statistical analyses were performed using Student t test. The heart of white mice contain diploid, tetraploid, octaploid, hexadecaploid (16 n) and 32 n cells. The amount of cardiomyocytes of different ploidy classes varies from one age group to another. The intensity of apoptosis increases with age. In all age groups the cardiomyocytes retain some capacity to synthesize DNA, which is not influenced by aging. However, the fact of DNA synthesis per se seems to be insufficient to drive adult cardiomyocytes through cell division. PMID- 16369075 TI - Argyrophilic nucleolar organizer regions in benign and malignant mesothelial lesions. AB - The aim our study was to assess the usefulness of AgNOR stain in distinguishing between benign and malignant mesothelial lesions. The patients were divided into three groups: group I -- reactive mesothelium (71 cases), group II -- hyperplastic mesothelium (66 cases), group III -- epithelial type mesothelioma (52 cases). Smears were stained by Pap and AgNOR methods. After staining, all cases were randomized for blind evaluation. Each case was viewed independently by two observers. AgNORs were identified as black, usually spheric particles observed within the nucleolus. For each cell, the number of AgNOR-positive cells and the number of AgNOR-dots per nucleus were counted. Our results show that AgNOR staining is useful to differentiate epithelial type mesothelioma and benign mesothelial lesions such as reactive and hyperplastic mesothelium. This differentiation is based primarily on the mean number of AgNOR-dots per cell rather than number of AgNOR-positive cells. AgNOR is highly sensitive, specific and cost-effective technology which can be used as an ancillary diagnostic approach for distinguishing between reactive and/or hyperplastic changes of mesothelium as well as in differential diagnosis of epithelial type mesothelioma. PMID- 16369076 TI - [Morphological changes of cardiovascular system at experimental hypertension, caused by no inhibition in condition of the laser irradiation of low-frequency]. AB - The aim of our study was the investigation of the morphological changes of cardiovascular system, caused by NO inhibition during the laser irradiation of low-frequency. On reception of L NAME and a low-frequency laser irradiation arterial pressure during all experiment remains within the norm. At slight increase of phonic arterial pressure the laser irradiation proved to be less effective, while on a background of high arterial pressure the irradiation does not interfere the further development of hypertension and persistency. Immediately after reception of L NAME and at a low-frequency laser irradiation, the fibrosis and inflammatory changes of myocardium have not been observed; on the background of the slight enhancement of arterial pressure, laser irradiation causes insignificantly expressed fibrosis, inflammatory changes in myocardium; while on the background of increased arterial pressure a laser irradiation doesn't interfere development of fibrosis, myocardium and inflammatory changes in it. Immediately after reception L NAME and at a laser irradiation of low frequency from the third day, a damage of cardiovascular walls, inflammatory changes, perivascular fibrosis have not been developed, while on the background of increased arterial pressure the irritation doesn't interfere the development of the above mentioned changes. Thus, the irradiation of a low-frequency laser might be recommended to prevent the development of hypertension and to avoid the complications of pre-clinical phase of hypertension. Though, the analogous recommendations are not foreseen for involvement in algorithm of the treatment of hypertension and correction of clinical evidence of the disease. PMID- 16369077 TI - [The mass transfer features differentiation in intestinal villi in morphogenetic process]. AB - The aim of this study was to investigate mass transfer features differentiation in intestinal villi in process of morphogenesis. The trial was carried out on 28 newborn white rats (weight 40-60 g) and on 24 adult rats (weight 190-220 g). There was used the method of scanning electron microscopy. It has been shown, that after the first nursing the amount of lipids sharply rises in intestinal villi. From the 7th day of life, there was reliable apical- basic gradient of lipids content in interstitium of intestinal villi. Lipids are absorbed and deported by the base of villi, from where they are sucked gradually by the initial lymphatics. However, the results confront with comparative data of lipid mass discrete density in interstitial tissue has shown resembling indices on all length of villi. Consequently, the point is, that with the age the width of villi base is getting larger, where the large amount of lipids may be located. PMID- 16369078 TI - [Structural indices of blood cells in young athletes after one month of training taking into account different types of sport]. AB - Blood cells of young sportsmen have been studied taking into account different types of sport (classical wrestling, judo, free-style wrestling). Our observations have shown that the most important shifts in blood cells take place during classical wrestling and judo. The most intensive changes were noted in erythrocytes and thrombocytes. The immune response shows different patterns in classical wrestling and judo. Data obtained using light microscope method are further supported by morphometric study of our clinical material. Thus, physical training plays an important role in the activation of functions of the organism, particularly in the activation of blood cells. Increase of euchromatin indicates to the activation of the nucleus, while the decrease, particularly in the cases of classical wrestling and judo, to the activation of perinuclear space and increase of interaction between the nucleus and cytoplasm. PMID- 16369079 TI - [Identification and hygienic assessment of irradiation doses of population in some districts of Samegrelo region]. AB - Internal and total irradiation doses for the population of Zugdidi, Khobi, Tsalenjikha districts (Samegrelo region, Georgia) have been defined and hygienically assessed; preventive measures have also been proposed for its reduction. Internal irradiation dose for the population of Samegrelo region was identified as 2.26 mSv/y, total irradiation dose -- 3.31 mSv/y, i.e. relatively increased. To reduce the total irradiation dose of the population apart from the need to prohibit use of high activity water for drinking and to provide population with new sources of low activity water, it is necessary to control and minimize doses caused by the artificial irradiation; this should primarily happen by means of regulation of medical radiological procedures and rational use of fertilizers with K-40 content. PMID- 16369080 TI - [Some peculiarities of vibration diseases in manganese mine workers]. AB - Vibration disease is one of the leading pathology in the structure of occupational diseases. The aim of our investigation was to study the peculiarities of vibration disease in manganese mine workers Georgia (dependence on the degree of disease, duration of vibration factors, the age and sex) in order to elaborate the complex of medico-preventive and rehabilitation measures. Results of the study showed that clinical picture of the disease was characterized by different damage syndromes of central, peripheral and vegetative nerve systems. On the basis of the investigation the complex of medico-preventive and rehabilitation measures was done for Chiatura manganese mines medical stations. PMID- 16369081 TI - [Vitamins C, B12 and folic acid in latent iron deficiency]. AB - The aim of the work was to study the level of vitamins C, B12 and folic acid in latent iron deficiency of different etiology (hipo- and anacid gastritis and menorrhagia). 81 patients with latent iron deficiency were investigated. Vitamin C levels were measured by refractometry, folic acid and vitamin B12 by radioimmune assay. The obtained results showed significant decrease of ascorbic acid and less apparent decrease of folic acid in the blood plasma. The content of vitamin B12 was unchanged. Decrease of vitamin C level is related to the changes of initial stages of iron metabolism and its further absorption. Less apparent changes of folic acid and vitamin B12 indicates, that such important stages of erithropoesis as DNA replication and cell proliferation are intact. Our results indicate to the development of metabolic disorders prior to revelation of iron deficiency anaemia, which need timely correction. PMID- 16369082 TI - Biorheological views of endothelial cell responses to mechanical stimuli. AB - Vascular endothelial cells are located at the innermost layer of the blood vessel wall and are always exposed to three different mechanical forces: shear stress due to blood flow, hydrostatic pressure due to blood pressure and cyclic stretch due to vessel deformation. It is well known that endothelial cells respond to these mechanical forces and change their shapes, cytoskeletal structures and functions. In this review, we would like to mainly focus on the effects of shear stress and hydrostatic pressure on endothelial cell morphology. After applying fluid shear stress, cultured endothelial cells show marked elongation and orientation in the flow direction. In addition, thick stress fibers of actin filaments appear and align along the cell long axis. Thus, endothelial cell morphology is closely related to the cytoskeletal structure. Further, the dynamic course of the morphological changes is shown and the related events such as changes in mechanical stiffness and functions are also summarized. When endothelial cells were exposed to hydrostatic pressure, they exhibited a marked elongation and orientation in a random direction, together with development of centrally located, thick stress fibers. Pressured endothelial cells also exhibited a multilayered structure with less expression of VE-cadherin unlike under control conditions. Simultaneous loading of hydrostatic pressure and shear stress inhibited endothelial cell multilayering and induced elongation and orientation of endothelial cells with well-developed VE-cadherin in a monolayer, which suggests that for a better understanding of vascular endothelial cell responses one has to take into consideration the combination of the different mechanical forces such as exist under in vivo mechanical conditions. PMID- 16369084 TI - Hemorheologic events in severe shock. AB - Persistent low perfusion and low blood pressure are the two major events in the pathogenesis of irreversible shock. This review is focused on our recent study on the mechanism of, and a new therapeutic approach to the two events in IS. One of the main causes of persistent low perfusion are leukocyte adhesion on venule walls and plugging in capillaries which comes from the low wall shear stress or shear rate, and high leukocyte-endothelial adhesion force in IS. However, blockade of leukocyte adhesion by monoclonal antibodies against the adhesion molecules can only attenuate the number of sticking WBC in venules, but cannot make an appreciable improvement in capillary reflow and survival rate in IS, because it is difficult for the agents to flow into an obstructed capillary. We have shown that the administration of Polydatin, a crystalline product isolated from a traditional Chinese medicine, can restore the pulse pressure with high survival rate in irreversible shock. With an increase in pulse pressure, and the highly dispersive force resulting from pulsatile blood flow, the stationary blood cells can be pushed away from the obstructed capillary and thus promote capillary reflow. Therefore, enhancement of pulse pressure is a key factor for the treatment of low perfusion in irreversible shock. Hyperpolarization of arteriolar smooth muscle cells occurs in irreversible shock, which inhibits the potential operated calcium channel and the influx of Ca2+ in arteriolar smooth muscle cells stimulated by norepinephrine, and finally leads to low vascular contractile responsiveness with refractory hypotension in irreversible shock. Activation of the potassium channels K(ATP) and BK(Ca) is involved in arteriolar smooth muscle cells hyperpolarization. In irreversible shock, ATP depletion, intracellular acidosis, ONOO- formation, and enhancement of a calcium spark results in activation of K(ATP) and BK(Ca) and consequent arteriolar smooth muscle cell hyperpolarization. Therefore, a new therapeutic strategy for refractory hypotension was suggested, including blockade of potassium channel activation to reconstitute vasoreactivity and the administration of vasopressors to elevate blood pressure in the treatment of irreversible shock. PMID- 16369083 TI - Catch bonds: physical models, structural bases, biological function and rheological relevance. AB - Force can shorten the lifetimes of macromolecular complexes (e.g., receptor ligand bonds) by accelerating their dissociation. Perhaps paradoxical at first glance, bond lifetimes can also be prolonged by force. This counterintuitive behavior was named catch bonds, which is in contrast to the ordinary slip bonds that describe the intuitive behavior of lifetimes being shortened by force. Fifteen years after their theoretical proposal, catch bonds have finally been observed. In this article we review recently published data that have demonstrated catch bonds in the selectin system and suggested catch bonds in other systems, the theoretical models for their explanations, possible structural bases, their relation to flow-enhanced adhesion, and the potential biorheological relevance. PMID- 16369085 TI - Volume flow estimation in the precapillary mesenteric microvasculature in vivo and the principle of constant pressure gradient. AB - Volume flow was estimated from axial erythrocyte velocity measurements in 30 mesenteric microvessels of 6 rabbits and was compared to Murray's law predictions. The diameters of capillaries and precapillary arterioles ranged between 5.6 and 12 microm. The significant pulsating flow component existing in these microvessels was taken into account by measuring instantaneous axial blood velocity throughout the course of a cardiac cycle and then averaging over the period. In addition, the effect of the velocity profile variation with diameter was taken into account, for the first time, by using a profile factor function. According to Murray's law, the relation between blood volume flow and diameter is governed by a 'cube' law. Curve fitting to volume flow and diameter data pairs for rabbits, showed a dependence of volume flow on diameter raised to the 4th power (with a correlation coefficient equal to 0.97). The above result supports the hypothesis that, in the precapillary part of microvasculature, the principle of constant longitudinal pressure gradient rather than the principle of minimum work may be valid. PMID- 16369086 TI - Numerical study of the impact of non-Newtonian blood behavior on flow over a two dimensional backward facing step. AB - Endothelial cell (EC) responsiveness to shear stress is essential for vasoregulation and plays a role in atherogenesis. Although blood is a non Newtonian fluid, EC flow studies in vitro are typically performed using Newtonian fluids. The goal of the present study was to determine the impact of non Newtonian behavior on the flow field within a model flow chamber capable of producing flow disturbance and whose dimensions permit Reynolds and Womersley numbers comparable to those present in vivo. We performed two-dimensional computational fluid dynamic simulations of steady and pulsatile laminar flow of Newtonian and non-Newtonian fluids over a backward facing step. In the non Newtonian simulations, the fluid was modeled as a shear-thinning Carreau fluid. Steady flow results demonstrate that for Re in the range 50-400, the flow recirculation zone downstream of the step is 22-63% larger for the Newtonian fluid than for the non-Newtonian fluid, while spatial gradients of shear stress are larger for the non-Newtonian fluid. In pulsatile flow, the temporal gradients of shear stress within the flow recirculation zone are significantly larger for the Newtonian fluid than for the non-Newtonian fluid. These findings raise the possibility that in regions of flow disturbance, EC mechanotransduction pathways stimulated by Newtonian and non-Newtonian fluids may be different. PMID- 16369087 TI - Effect of enhanced red blood cell aggregation on blood flow resistance in an isolated-perfused guinea pig heart preparation. AB - The role of red blood cell (RBC) aggregation as a determinant of in vivo blood flow is still unclear. This study was designed to investigate the influence of a well-controlled enhancement of RBC aggregation on blood flow resistance in an isolated-perfused heart preparation. Guinea pig hearts were perfused through a catheter inserted into the root of the aorta using a pressure servo-controlled pump system that maintained perfusion pressures of 30 to 100 mmHg. The hearts were beating at their intrinsic rates and pumping against the perfusion pressure. RBC aggregation was increased by Pluronic (F98) coating of RBC at a concentration 0.025 mg/ml, corresponding to about a 100% increment in RBC aggregation as measured by erythrocyte sedimentation rate. Isolated heart preparations were perfused with 0.40 l/l hematocrit unmodified guinea pig blood and with Pluronic coated RBC suspensions in autologous plasma. At high perfusion pressures there were no significant differences between the flow resistance values for the two perfusates, with differences in flow resistance only becoming significant at lower perfusion pressures. These results can be interpreted to reflect the shear dependence of RBC aggregation: higher shear forces associated with higher perfusion pressures should have dispersed RBC aggregates resulting in blood flow resistances similar to control values. Experiments repeated in preparations in which the smooth muscle tone was inhibited by pre-treatment with papaverine indicated that significant effects of enhanced RBC aggregation could be detected at higher perfusion pressures, underlining the compensatory role of vasomotor control mechanisms. PMID- 16369088 TI - Current state and prospects of macromolecular crystallography. AB - The current situation and possible future development of macromolecular crystallography are reviewed. The rapid progress and maturation of this field in recent years have to a large extent been made possible by the inspiration and generous support of several active structural genomics initiatives. Two tendencies can be currently observed: one which treats protein crystallography as a highly automatic tool for investigating various biological problems without the need to engage in the intricacies of the technique and a second approach where this method is applied to crystals of difficult, large and complex biological systems, requiring a deeper knowledge of various aspects of crystallography. In the near future it is expected that these two trends will coexist, developing in a parallel fashion. PMID- 16369089 TI - Instrumentation for synchrotron-radiation macromolecular crystallography. AB - X-ray diffraction is an extremely important tool for structure determination of biological macromolecules, to the extent that currently around 85% of Protein Data Bank entries result from X-ray measurements. Many of these structure determinations use synchrotron radiation for data collection. This article aims to give synchrotron users an overview of the functioning of a synchrotron beamline and how the performance of various instruments combines to allow the collection of diffraction data. PMID- 16369090 TI - Combinatorial library approaches for improving soluble protein expression in Escherichia coli. AB - High-throughput screening methodologies are already used in structural biology to define efficient protein crystallization and expression conditions. Recently, screening approaches have been extended to the optimization of genetic constructs for improved soluble protein expression. With similarities to the directed evolution strategies used in protein engineering, a target gene encoding a poorly expressed protein is mutated by truncation, fragmentation or point mutation. Rare clones with improved protein expression characteristics are then isolated from the random library using a phenotypic screen or selection. This article reviews the progress in this field and provides a general overview of relevant mutation methods, screens and selections. PMID- 16369091 TI - A review of techniques for maximizing diffraction from a protein crystal in stilla. AB - There are a number of published reports of techniques that may extend the diffraction limits or otherwise improve the quality of the X-ray diffraction data from a crystal. This review touches on some of these techniques to provide a way forward for the experimentalist burdened with less than ideal crystals. PMID- 16369092 TI - Cryocooling and radiation damage in macromolecular crystallography. AB - Advances in cryocrystallographic techniques for macromolecular crystallography have been intimately intertwined with efforts to reduce the deleterious effects of X-ray damage inflicted during the collection of diffraction data. A brief overview of cryomethods and their rationale is given. This is followed by a summary of our current limited understanding of radiation damage in cryocooled crystals, investigations aimed at minimizing its effects and finally some developments which actually utilize it both for phasing and to extend structural knowledge. PMID- 16369093 TI - The integration of macromolecular diffraction data. AB - The objective of any modern data-processing program is to produce from a set of diffraction images a set of indices (hkls) with their associated intensities (and estimates of their uncertainties), together with an accurate estimate of the crystal unit-cell parameters. This procedure should not only be reliable, but should involve an absolute minimum of user intervention. The process can be conveniently divided into three stages. The first (autoindexing) determines the unit-cell parameters and the orientation of the crystal. The unit-cell parameters may indicate the likely Laue group of the crystal. The second step is to refine the initial estimate of the unit-cell parameters and also the crystal mosaicity using a procedure known as post-refinement. The third step is to integrate the images, which consists of predicting the positions of the Bragg reflections on each image and obtaining an estimate of the intensity of each reflection and its uncertainty. This is carried out while simultaneously refining various detector and crystal parameters. Basic features of the algorithms employed for each of these three separate steps are described, principally with reference to the program MOSFLM. PMID- 16369094 TI - A quantitative approach to data-collection strategies. AB - Statistical descriptors of the X-ray diffraction data set for a macromolecular crystal can be modelled using the information present in the initial diffraction images. Quantitative relationships between the crystal quality, beam apertures, oscillation width, resolution limit, redundancy and the data statistics are presented. They are analysed in terms of the radiation-dose requirements based on modelling in program the BEST. The influence of radiation damage on the data statistics is discussed. PMID- 16369095 TI - The care and nurture of undulator data sets. AB - Undulator radiation is the X-ray source of choice for modern macromolecular crystallography beamlines. Here, the basic properties of undulator sources are described and it is indicated why they make such good X-ray sources for macromolecular crystallography. Collection of excellent data from these beamlines is not always straightforward; therefore, a number of rules are postulated for undulator data collection and guidelines are offered which will help to ensure a satisfactory experiment. PMID- 16369096 TI - Scaling and assessment of data quality. AB - The various physical factors affecting measured diffraction intensities are discussed, as are the scaling models which may be used to put the data on a consistent scale. After scaling, the intensities can be analysed to set the real resolution of the data set, to detect bad regions (e.g. bad images), to analyse radiation damage and to assess the overall quality of the data set. The significance of any anomalous signal may be assessed by probability and correlation analysis. The algorithms used by the CCP4 scaling program SCALA are described. A requirement for the scaling and merging of intensities is knowledge of the Laue group and point-group symmetries: the possible symmetry of the diffraction pattern may be determined from scores such as correlation coefficients between observations which might be symmetry-related. These scoring functions are implemented in a new program POINTLESS. PMID- 16369097 TI - Intensity statistics in twinned crystals with examples from the PDB. AB - Entries deposited in the Protein Data Bank as of February 2004 for which both model and X-ray data were available were analysed to identify cases of twinning using such simple statistics as the R factor between potential twin-related reflections. Careful consideration of all identified twins showed that in many cases twinning was ignored during structure solution and refinement. Manual analysis of the models showed that twinning often occurs in association with rotational pseudosymmetry parallel to the twinning operator. The coexistence of these two phenomena complicates the detection and diagnostics of twinning using currently available twinning tests. It was concluded that a twinning-detection step should be incorporated in every stage of structure analysis from data acquisition to refinement and validation. PMID- 16369098 TI - Some aspects of quantitative analysis and correction of radiation damage. AB - Radiation damage is the major source of systematic error in macromolecular data collected at third-generation synchrotron beamlines. In this paper, a simple way of analysing data for radiation damage is proposed and shown to give results that are easy to interpret. Results of correction of radiation damage obtained with an exponential decay function as implemented in XSCALE (from the XDS package) are shown, and aspects of the mathematical treatment of radiation damage, as well as experimental requirements for the correction and utilization of radiation damage are discussed. Furthermore, a method for quantifying the coverage and evenness of sampling of rotation range is proposed. PMID- 16369099 TI - Industrial perspective on X-ray data collection and analysis. AB - Protein crystallography methods applied by research teams in the pharmaceutical industry to support the process of discovery of new medicines are not greatly different from those used by academic structural biology groups. However, owing to the specific aims of the pharmaceutical industry, the approaches and working practices are often quite distinct. This applies to both the determination of novel structures of drug targets and complexes of these targets with potential drugs. To make any significant impact on ongoing medicinal chemistry projects, crystal structures have to be delivered on time and must provide answers to specific questions. Owing to the high number of crystal structures typically solved by industrial research groups, development of technology and computational methods which speed up the process and increase throughput is of vital importance. This paper presents an overview of current approaches to X-ray data collection and processing within the industrial environment, with examples of how technology is used to address the challenges structural biology faces in this "high-throughput-everything" period. PMID- 16369100 TI - Structure determination of human semaphorin 4D as an example of the use of MAD in non-optimal cases. AB - Semaphorins are an important class of signalling molecules involved in axon guidance, immune function and angiogenesis. They are characterized by having an extracellular sema domain of about 500 residues. The steps involved in the determination of the structure of human semaphorin 4D are described here as a case study of selenium MAD phasing in a difficult case with low symmetry, moderate diffraction and low selenium content. A particular feature of this study was the large number of diffraction images required to give data of sufficient quality for structure determination and these data are re-analyzed here to investigate the effects of radiation damage on eventual data quality and to suggest strategies for successful MAD phasing in similar difficult cases. PMID- 16369101 TI - Entropy and surface engineering in protein crystallization. AB - Protein crystallization remains a key limiting step in the characterization of the atomic structures of proteins and their complexes by X-ray diffraction methods. Current data indicate that standard screening procedures applied to soluble well folded prokaryotic proteins yield X-ray diffraction crystals with an approximately 20% success rate and for eukaryotic proteins this figure may be significantly lower. Protein crystallization is predominantly dependent on entropic effects and the driving force appears to be the release of ordered water from the sites of crystal contacts. This is countered by the entropic cost of ordering of protein molecules and by the loss of conformational freedom of side chains involved in the crystal contacts. Mutational surface engineering designed to create patches with low conformational entropy and thereby conducive to formation of crystal contacts promises to be an effective tool allowing direct enhancement of the success rate of macromolecular crystallization. PMID- 16369102 TI - Genetic and environmental factors associated with the development of hypertension in pregnancy. AB - Hypertension in pregnancy (HP), one of the most common causes of perinatal deaths, is a multifactorial disease with genetic and environmental factors involved in its etiology. We have carried out molecular epidemiologic research with the purpose of (1) identifying gene variants associated with HP in Japanese women, and (2) analyzing the genetic and environmental factors involved in the pathophysiology of the disease. Self-administered questionnaires were returned by the subjects between 1 and 6 months after delivery. The candidate genetic variants were identified by use of a PCR-RFLP method. T235 of AGT, C1166 of AT1 and Asp298 of NOS3 were respectively associated with HP, although no significant associations were found between the common genetic variants and HP in ACE, FV, MTHFR, B3AR, TNF-A, PAI-1, GSTP1, mEH, and LPL. In analyses using genetic, environmental and lifestyle factors, 5 factors before pregnancy and 4 factors during pregnancy were significantly associated with HP in univariate analysis. Further multivariate analysis revealed 3 factors before pregnancy, i.e. "prepregnancy BMI > or = 24 kg/m(2)", "family history of hypertension" and "TT genotype of AGT", and 2 factors during pregnancy, i.e. "mentally stressful condition" and "salty dishes preferred". Dividing the subjects into 2 subgroups according to whether they possessed "TT genotype of AGT" or not, we identified acquired risk factors before and during pregnancy for HP in each groups. The multivariate analysis identified "mentally stressful condition" as a potent significant risk factor during pregnancy in the former subgroup. However, there were no significant risk factors concerning and "mental stress" in the latter subgroup. Through further exploration of the risk factors associated with HP, we hope to provide useful suggestions about the development of new and effective preventive measures for a range of multifactorial diseases. PMID- 16369103 TI - Epidemiologic pictures of Kawasaki disease in Shanghai from 1998 through 2002. AB - BACKGROUND: Epidemiologic features of Kawasaki disease in China is still not clear. METHODS: A questionnaire form and diagnostic guidelines for Kawasaki disease were sent to hospitals in Shanghai, which provided with pediatric medical care. All patients with Kawasaki disease diagnosed during January 1998 through December 2002 were recruited in this study. RESULTS: A total of 768 patients with Kawasaki disease were reported. The incidence rates of Kawasaki disease for each year were 16.79 (1998), 25.65 (1999), 28.16 (2000), 28.05 (2001), and 36.76 (2002) per 100,000 children under 5 years of age. The male/female ratio was 1.83:1. The age at onset ranged from 1 month to 18.8 years (median: 1.8 years). The disease occurred more frequently in spring and summer. Fever was the most common clinical symptom, followed by oral changes, extremities desquamate, rash, conjunctive congestion, lymphadenopathy, extremities swelling, and crissum desquamate. Cardiac abnormalities were found in 24.3% of patients. The most common cardiac abnormality was coronary artery lesions including dilatation (68%) and aneurysm (10%). The case-fatality rate at acute stage of the disease was 0.26%. A second onset of the disease occurred in 1.82% of patients. CONCLUSIONS: The incidence rate of Kawasaki disease in Shanghai is lower than that reported in Japan, but higher than those in western countries. The increasing trend in incidence, sex distribution and cardiac abnormalities are similar to those in previous reports. The seasonal distribution is similar to the report from Beijing and different from other reports. PMID- 16369104 TI - The value of combining serum alanine aminotransferase levels and body mass index to predict mortality and medical costs: a 10-year follow-up study of National Health Insurance in Shiga, Japan. AB - BACKGROUND: Evidence suggests that the predictive value of serum alanine aminotransferase (ALT) levels for prognosis, measured by indices such as all cause mortality and medical costs, may be modified by body mass index (BMI). However, the relationship between serum ALT and BMI has not been satisfactorily elucidated. METHODS: Four thousand, five hundred and twenty-four community dwelling Japanese National Health Insurance beneficiaries, 40-69 years old, were classified into five categories according to their serum ALT levels (IU/L) (ALT<20, 20< or =ALT<30, 30< or =ALT<40, 40< or =ALT<50 and 50< or =ALT) and followed for 10 years. Hazard ratios for all-cause mortality, with reference to the lowest serum ALT category, and medical costs per person were evaluated for each serum ALT category after analyzing interactions between serum ALT levels and BMI for all-cause mortality and for medical costs. RESULTS: A significant interaction between serum ALT levels and BMI was observed. In participants below the median BMI, positive, graded relationships were identified between serum ALT levels and all-cause mortality as well as between serum ALT levels and personal medical costs. The multivariate-adjusted hazard ratio in the "50< or =ALT" category showed an approximately 8-fold increase. However, in the participants at or above the median BMI, no significant relationships between serum ALT levels and all-cause mortality or personal medical costs were identified. CONCLUSIONS: In these Japanese participants, the predictive value of serum ALT levels for prognosis was more evident if BMI was taken into account. A combination of high serum ALT levels and below median BMI was associated with excess mortality and medical costs. PMID- 16369105 TI - Factors associated with functional limitation in stair climbing in female Japanese patients with knee osteoarthritis. AB - BACKGROUND: Osteoarthritis (OA) of the knee is a common form of arthritis, and affects quality of life. We investigated factors associated with functional limitation in stair climbing among female Japanese patients with knee OA. As weight is a known risk factor for knee OA, we focused on body weight at 40 years of age, and examined the association with present weight, past weight, and weight change. METHODS: Subjects were 360 Japanese women aged 40-92 years who were newly diagnosed with knee OA at 3 university hospitals over a 1-year period. Factors associated with the severity of functional limitation in stair climbing were assessed by calculating odds ratios (OR) using the proportional odds model in logistic regression. RESULTS: Weight at diagnosis showed a positive association with severe functional limitation in stair climbing; however, a negative association was observed for weight change since age 40. Further analysis indicated that the association with weight at age 40 (highest vs. lowest quartile, OR=2.84, 95% confidence interval: 1.03-7.83, trend p=0.071) is stronger than weight at diagnosis. Other significant characteristics were age (70+ vs. 40 59 years, OR=7.37), previous knee pain and/or swelling 12 years or more before diagnosis (OR=2.67), and physical work (OR=1.94). In addition, higher parity was found to be a negatively associated factor (for tripara or more, OR=0.41). CONCLUSIONS: This study identified factors, such as heavy weight at age 40 and physical labor, which are potentially useful for preventing severe functional limitation for female knee OA patients. In addition, higher parity was associated with milder stair climbing limitation. PMID- 16369106 TI - Antimitochondrial antibody negative primary biliary cirrhosis in Japan: utilization of clinical data when patients applied to receive public financial aid. AB - BACKGROUND: We examined patients who showed laboratory and histological evidence of primary biliary cirrhosis (PBC) in the absence of antimitochondrial antibody (AMA) to elucidate the characteristics of AMA negative PBC. METHODS: From a total of 5,805 patients with symptomatic PBC, 2,419 cases (41.7%) were selected in the present study, who were diagnosed using the following criterion; chronic non suppurative destructive cholangitis was histologically observed and laboratory data did not contradict PBC. The information collected from records included sex, age, symptoms, physical findings, and complicated autoimmune diseases. We then evaluated these data according to the positivity of AMA. RESULTS: Of the total subjects, 470 cases (19.4%) were found to be negative for AMA. The proportion of female patients was higher among the AMA negative group than among the AMA positive one. Pruritus was found less frequently among patients with AMA negative PBC than among those with AMA positive PBC. Levels of alkaline phosphatase,gamma glutamyl transpeptidase, and IgM were significantly lower among patients with AMA negative PBC than among those with AMA positive PBC. Complications such as Sjogren's syndrome, rheumatoid arthritis, and scleroderma, including CREST syndrome, were found with significantly higher frequency among patients with AMA negative PBC than among those with AMA positive PBC. CONCLUSION: Considering serum level of IgM and frequencies of complicated autoimmune diseases, it is possible that Japanese patients with AMA negative PBC are consistent with the disease entity of autoimmune cholangitis reported in western countries. PMID- 16369107 TI - Comprehensive health education combining hot spa bathing and lifestyle education in middle-aged and elderly women: one-year follow-up on randomized controlled trial of three- and six-month interventions. AB - BACKGROUND: This study attempted to clarify the duration of effects of 3- and 6 month comprehensive health education programs based on hot spa bathing, lifestyle education and physical exercise for women at 1-year follow-up. METHODS: We examined middle-aged and elderly women who were randomly divided into two groups and followed up them for one year. Spa programmers instructed subjects for one hour in lifestyle education and physical exercise and for one hour in a half bath (salt spring, temperature at 41.5 degrees C) once a week. The program for the 3 month group (n=19) was repeated in the 6-month group (n=14). The evaluation items were body mass index, PWC75%HRmax (by a bicycle ergometer as aerobic capacity), blood profiles (total cholesterol, HDL cholesterol, arteriosclerotic index, uric acid, and hemoglobin A1c), profile of mood states, self-rating depression scale, subjective happiness, pains in the knee and back, and active modification of lifestyle. RESULTS: There were significant interactions between groups and response over time to aerobic capacity, hemoglobin A1c, back pain, vigor, fatigue and self-rating depression (respectively, p<0.05). Duration of effects was longer for the 6-month intervention than for the 3-month intervention. CONCLUSIONS: Beneficial effects of 6-month intervention on hemoglobin A1c, aerobic capacity, pains in the back, vigor, fatigue and depression remained significant at the 1 year follow-up. Duration of effects was longer in the 6-month intervention than in the 3-month intervention. PMID- 16369108 TI - Genotype announcement to Japanese smokers who attended a health checkup examination. AB - BACKGROUND: Genotype announcement may be one of the effective methods to induce smoking cessation, but the studies are limited throughout the world. METHODS: Subjects were smokers who attended a health checkup examination provided by a local government in Hokkaido, Japan, 2003. Those who agreed to know their genotypes were informed of the genotypes of glutathione S-transferease (GST) M1 present/null, GSTT1 present/null, and NAD(P)H:quinone oxidoreductase 1 (NQO1) C609T (Pro187Ser). RESULTS: Out of 143 smokers (92 males and 51 females), 101 individuals participated in the present study. A postal questionnaire one year after the genotype announcement found that 8 persons (6 males and 2 females) of 41 respondents had quitted smoking. Two of 8 quitters stated that they had quitted smoking due to the announcement. There were none who regretted the genotype tests. CONCLUSION: Although the cessation rate, 7.9% (8/101) at least, was not marked, no harmful effects were observed among the respondents. PMID- 16369109 TI - Effects of concentrated ambient particles on heart rate variability in spontaneously hypertensive rats. AB - In the present study, the cardiovascular toxicity of PM(2.5) was determined in spontaneously hypertensive (SH) rats using the standard deviation of normal-to normal intervals (SDNN) and root mean square of successive differences of adjacent normal-to-normal intervals (RMSSD) as outcome measurements. Four SH rats implanted with radiotelemetry transmitters were repeatedly exposed to concentrated PM(2.5) in nose-only exposure chambers. Gravimetric analysis revealed the mean post-concentrating mass concentration of particles during the 5 h of exposure was 202 mug/m(3). Using each animal as its own control and linear mixed-effects model, to adjust for circadian nature and individual differences, we found that SDNN decreased by 15% initially then gradually decreased to 60% of the initial value at the end of exposure. Our results indicate that concentrated PM(2.5) may decrease SDNN on SH rats during PM exposure. The study also showed that SDNN is more sensitive to PM induced effects than RMSSD. PMID- 16369110 TI - Relationships between work-related factors and disorders in the neck-shoulder and low-back region among female and male ambulance personnel. AB - This cross-sectional study on a random sample of 1,500 ambulance personnel investigated the relationships between self-reported work-related physical and psychosocial factors, worry about work conditions, and musculoskeletal disorders among female and male ambulance personnel. Three different outcomes, complaints, activity limitation, and sick leave, for the neck-shoulder and low-back region, respectively, were chosen. Among the female personnel, physical demands was significantly associated with activity limitation in the neck-shoulder (OR 4.13) and low-back region (OR 2.17), and psychological demands with neck-shoulder (OR 2.37) and low-back (OR 2.28) complaints. Among the male personnel, physical demands was significantly associated with low-back complaints (OR 1.41) and activity limitation (OR 1.62). Psychological demands and lack of social support were significantly associated with neck-shoulder complaints (OR 1.86 and OR 1.58, respectively) and activity limitation (OR 3.46 and OR 1.71) as well as activity limitation due to low-back complaints (OR 2.22 and OR 1.63). Worry about work conditions was independently associated with activity limitation due to low-back complaints among the female (OR 5.28), and to both neck-shoulder and low-back complaints (OR 1.79 and OR 2.04, respectively) and activity limitation (OR 2.32 and OR 1.95) among the male personnel. In conclusion, the association patterns between physical and psychological demands and MSDs suggest opportunities for intervention. PMID- 16369111 TI - A prospective study on the influences of workplace stress on mental health. AB - The purpose of this study was to clarify the influences of subjective stress in the workplace on mental health according to work contents in subjects without mental health problems. In a survey in fiscal 1997, 1,246 (84.6%) of all 1,363 workers of a manufacturing company in Japan, responded to a questionnaire including questions on subjective stress and GHQ60, and 1,135 workers answered all questions (effective response rate, 83.3%). In fiscal 1997, there were 705 "mentally healthy" workers with a GHQ score of 16. Of the 705 workers, 603 (85.5%) could be followed up by another survey 2 yr later (fiscal 1999). According to the contents of the work, multiple logistic regression analysis was performed using the status of mental health in fiscal 1999 as a dependent variable and subjective workplace stress, sex, age, smoking, and alcohol usage in fiscal 1997 as explanatory variables. The statistically significant items relating to problematic mental health for "factory work" were "too much competition", "feel pressed for time", and "cannot keep up with new technology" (Table 4). For "too much competition", the odds ratios (95% CI) of "always" and "sometimes" to "do not feel" were 4.04 (1.39-11.76) and 1.85 (0.92-3.70), respectively. For "feel pressed for time", the odds ratios (95% CI) of "always" and "sometimes" to "do not feel" were 2.40 (1.08-5.35) and 0.98 (0.46-2.06), respectively. For "cannot keep up with new technology", the odds ratio (95% CI) of "always" and "sometimes" to "do not feel" were 6.54 (0.37-116.36) and 2.52 (1.34-4.77), respectively. The statistically significant items relating to problematic mental health for "research and development or office work" were "too much trouble at work" and "feel pressed for time". For "too much trouble at work", the odds ratios (95% CI) of "always" and "sometimes" to "do not feel" were 3.92 (1.41-10.93) and 1.44 (0.67-3.09), respectively. For "feel pressed for time", the odds ratios (95% CI) of "always" and "sometimes" to "do not feel" were 2.69 (1.04-6.93) and 2.09 (0.94-4.67), respectively. Our study suggests that subjective stress items aggravating mental health are different among work contents. If we could identify items of workplace stress influencing mental health by work contents, we could utilize the results for a mental health promotion program. PMID- 16369112 TI - A cross-sectional study of alcohol drinking and health-related quality of life among male workers in Japan. AB - BACKGROUND: Although light and moderate alcohol drinkers are likely to have better subjective health, the sub-scales for subjective health have not been well documented. METHODS: We studied 4,521 male workers aged 25 yr and older with no history of cancer or cardiovascular disease, in 12 occupational groups in Japan. Data were from the High-risk and Population Strategy for Occupational Health Promotion Study (HIPOP-OHP). Drinking status was classified according to daily alcohol intake or frequency of drinking. We assessed the health-related quality of life (HRQOL) based on scores for five scales of the SF-36. RESULTS: Decreased odds ratios of sub-optimal HRQOL conditions, defined as less than the median SF 36 scores, for Role-Physical and General Health were found among persons who consumed 1.0 to 22.9 g/d of alcohol. Odds ratios for sub-optimal Vitality conditions were lowered according to increased levels of alcohol intake. Role Emotional scores were not associated with alcohol drinking. People who drank 5 to 6 d/wk had higher levels of Role-Physical and Vitality, and those who drank 1 to 2 d/wk had better Vitality and Mental Health scores than non-drinkers. When adjusted for age, marital status, working hours, physical activity at work, self reported job stress, smoking, regular exercise, hypertension, hyperlipidemia, and diabetes, the associations were almost unchanged except for General Health. CONCLUSIONS: Associations of drinking patterns with subjective health varied in five sub-scales of the SF-36. Overall, alcohol drinkers rated their health as good in comparison with non-drinkers. PMID- 16369113 TI - Expression of clara cell secretory protein in the lungs of rats exposed to crystalline silica in vivo. AB - It has been theorized that Clara cell secretion protein (CCSP) plays a critical role in regulating the acute inflammatory response in the lung. We hypothesized that CCSP is also related to lung injury induced by occupational dust. The present study was conducted to investigate the time course of the expression of CCSP in lungs exposed to crystalline silica in vivo. Male Wistar rats were administered 1 mg or 2 mg of silica suspended in saline by a single intratracheal instillation and were sacrificed at 3 d, 1 wk, 1 month, 3 months and 6 months of recovery time. The expression of CCSP was observed by RT-PCR and western blot analysis. Exposure to 2 mg of silica decreased in levels of CCSP mRNA at 3 d, 1 wk, 1 month and 6 months following intratracheal instillation. The protein level of CCSP in silica-exposed rats was decreased at 3 d, 7 d and 1 month after a single instillation of 2 mg. The decreases in CCSP at the acute phase in this experiment suggest that CCSP may regulate the acute injury of the lung exposed to silica. PMID- 16369114 TI - A Prospective Cohort Study of Employment Status and Mortality from Circulatory Disorders among Japanese Workers. AB - This study prospectively examined the association between employment status (employed or self-employed) and the risk of death from circulatory diseases among Japanese workers. A baseline survey was conducted between 1988 and 1990 among 110,792 inhabitants of 45 areas. Follow-up surveys were conducted annually and causes of death were identified from death certificates. Analysis was restricted to 25,945 individuals (15,434 male and 10,511 female) with ages ranging from 40 to 59 years. These subjects were employed or self-employed at the time of recruitment. The risks of self-employment for death due to circulatory system disease, ischemic heart disease and cerebrovascular disease were estimated using the Cox proportional hazards model. During the 10-year follow-up period (151,817 and 104,870 person-years for males and females, respectively), 720 male and 193 female deaths were recorded. No significant differences were detected between the employed and self-employed workers in the total death risk, or the risk of death from ischemic heart disease. However, self-employed men showed a significantly lower risk of death from cerebrovascular disease compared with employed men (relative risk=0.58; 95% confidence interval=0.35, 0.97). Our findings suggest that employed men are at increased risk of death from cerebrovascular disease compared with self-employed men. PMID- 16369115 TI - Exposure to lead, cadmium and chromium among spray painters in automobile body repair shops. AB - Environmental and biological monitoring of lead, cadmium and chromium levels in spray painters is reported. All of the study subjects worked in automobile body repair shops that had no standard spraying room. They were divided into 2 groups, those who wore an aerosol-removing respirator while spraying (n=20) and those who did not wear the respirator (n=50). Air in the breathing zone of each subject was sampled and analyzed for lead, cadmium and chromium levels. The subjects' blood lead levels and urinary cadmium and chromium levels were also measured along with those of a control group. The mean environmental and biological levels of these metals between the two groups of the painters were not significantly different (p>0.05). However, the biological levels of the metals were significantly higher in the study groups than in the control group (p<0.01). On-site observations revealed that improper use of an aerosol-removing respirator, lack of an isolated spraying room and poor personal hygiene habits caused the failure to prevent heavy metal exposure among the automobile spray painters. PMID- 16369116 TI - The health of volunteer firefighters three years after a technological disaster. AB - On May 13, 2000, a firework depot exploded in a residential area of the city of Enschede, The Netherlands. Many disaster workers responded, including volunteer firefighters, a group that has received little attention in disaster research. This study examined the presence of health problems in volunteer firefighters who were involved in disaster work, three years after the disaster. Furthermore, it was investigated whether demographic characteristics and disaster exposure predicted health problems. The study population consisted of 246 volunteer firefighters who were deployed in disaster work and 71 non-deployed controls. These firefighters completed a questionnaire which inquired about their perceived health and health change, physical symptoms, post-traumatic stress, mental health problems, and health care utilization. Three years after the disaster, no health differences emerged between deployed and non-deployed firefighters. Good health and health improvement over the previous year were reported. Respondents who encountered more distressing experiences during disaster work or carried out more direct disaster-related recovery tasks reported more mental health problems and health care utilization. However, the most reliable predictors of health problems were distressing experiences unrelated to the disaster in personal life or during work. Three years after the disaster, the health of volunteer firefighters involved in the disaster work was not much impaired, possibly because aftercare was available and utilized. Nevertheless, disaster exposure was associated with elevated post-traumatic stress symptoms and mental health care utilization. Health care workers should direct specific attention to the treatment and prevention of post-traumatic stress symptoms in cases of major accidents or disasters. PMID- 16369117 TI - Occupational safety and health aspects of corporate social responsibility (CSR) in Japanese companies listed on the Tokyo Stock Exchange (TSE) first section. AB - Recently, corporate social responsibility (CSR) is becoming widely recognized as an issue for Japanese companies. Corporate responsibility for employees is considered important by various stakeholders, and occupational safety and health is regarded as one of these responsibilities. The present authors examined this issue from the viewpoint of corporate management by analysis of statements found in CSR-related reports. For companies listed on the First Section of the Tokyo Stock Exchange (TSE), we searched for CSR-related reports, and titles and contents, based on two established guidelines: the GRI Sustainability Reporting Guidelines 2002 as the international reference and the Environmental Reporting Guidelines of the Ministry of the Environment, Government of Japan, as the domestic reference. Corporations that published CSR reports were 26.3% (416/1,581) of the total, and large differences were recognized by type of industry. Comparing the numbers of pages for various contents, more concern was shown about the environment than about social activity, indicating the environment to be the main issue of CSR in Japan. In the items included in the guidelines, many matters about occupational accidents were mentioned, but it was found that statements regarding HIV/AIDS, which is not of such strong social concern in Japan, and statements regarding the costs of safety that are difficult to calculate were few. However, statements regarding mental health, which is of high interest socially, were many, even though this issue is not included in the two guidelines used. In revising the guidelines, these matters should be reviewed. In the future, continuance of analysis of CSR-related reports with regard to changes and comparisons with overseas reports will help improve occupational safety and health. PMID- 16369118 TI - Occupational exposure to volatile organic compounds and mitigation by push-pull local exhaust ventilation in printing plants. AB - The extensive use of multiple organic solvents in offset lithographic printing causing high emissions of volatile organic compounds (VOCs) indeed poses a serious risk to printing workers' health. In this study, indoor air quality (IAQ) assessments were carried out in seven printing plants and the main objectives were to understand the effect of VOC emissions on IAQ and develop effective mitigation measures to protect workers. The thorough gas chromatography/mass spectrometry (GC/MS) measurements showed that although a variety of VOCs were presented in the indoor air, none of them was found close to individual 8-h time weighted average (TWA) of the occupational exposure limit (OEL). The additive effect was also found below the critical value of unity. However, short-term personal exposure to total volatile organic compounds (TVOCs) was exceedingly high when a print worker carried out blanket and ink roller cleaning procedures. Therefore, the occupational health risk was mainly due to repeated short-term exposures during intermittent VOC-emitting procedures rather than long-term exposure to background VOCs. Push-pull local exhaust ventilation (LEV) was identified as an effective mitigation measure. Computational fluid dynamics (CFD) analysis was conducted to study the push-pull LEV operation. It was found that there existed a threshold LEV air flow rate for an abrupt reduction in the worker's exposure to VOCs. The reduction was less sensitive when the LEV airflow was further increased beyond the threshold. These phenomena, consistent with experimental results reported by other investigators, were explained by detailed CFD analysis showing the competition between the general ventilation and the push pull LEV to become the dominating driving force for the resultant local flow pattern. PMID- 16369119 TI - Association between beta2-adrenergic receptor genetic polymorphisms and total serum IgE in asthmatic patients of Chinese Han nationality. AB - BACKGROUND: beta(2)-Adrenergic receptor (beta(2)-AR) polymorphisms occurring at amino acid position 16 (Arg-Gly) and 27 (Gln-Glu) are known to be functionally relevant and also disease-modifying in subjects with asthma. It has been found in Caucasoid asthmatic patients that the Gln27 genotype beta(2)-AR was associated with an increase in total serum IgE levels. The association between beta(2)-AR genetic polymorphisms and total serum IgE in asthmatic patients of Chinese Han nationality remains to be established. OBJECTIVES: It was the aim of this study to investigate the association between beta(2)-AR genetic polymorphisms and total serum IgE in asthmatic patients of Chinese Han nationality. METHODS: All 59 asthmatic patients investigated (27 males and 32 females, aged between 16 and 60 years) were people of Chinese Han nationality. They were tested for their total serum IgE levels with the enzyme-linked immunosorbent assay test, and beta(2)-AR genetic polymorphisms were tested with the polymerase chain reaction allele specific oligonucleotide hybridization assay. RESULTS: There was a significant difference of serum IgE levels among three beta(2)-AR 27 loci groups (p < 0.0001), with the highest IgE level [(1.24 +/- 0.25) x 10(6) IU/l] in the Gln/Gln group and the lowest IgE level [(0.48 +/- 0.06) x 10(6) IU/l] in the Glu/Glu group. No polymorphism of beta(2)-AR 16 loci was found to be associated with total serum IgE (p > 0.05). CONCLUSIONS: Our research suggested that in asthmatic patients of Chinese Han nationality, the beta(2)-AR genetic polymorphism at 27 loci could be associated with serum IgE levels and it might therefore play an important role in the determination of phenotypes of bronchial asthma. PMID- 16369120 TI - Association between beta2-adrenergic receptor genetic polymorphisms and nocturnal asthmatic patients of Chinese Han nationality. AB - BACKGROUND: As a result of the finding that the mutation of Arg into Gly at beta(2)-adrenergic receptor (beta(2)-AR)16 loci could promote the downregulation effect triggered by the beta(2)-agonist, it was supposed that Gly16 might be associated with the downregulation of beta(2)-AR in patients with nocturnal asthma. OBJECTIVE: It was the aim of this study to analyze the association between beta(2)-AR genetic polymorphisms and nocturnal asthmatic patients of Chinese Han nationality. METHODS: A polymerase chain reaction allele-specific oligonucleotide hybridization assay was used to determine 16 and 27 loci alleles of beta(2)-AR genetic polymorphisms in 25 nocturnal asthmatic patients (nocturnal asthma group), 22 non-nocturnal asthmatic patients (non-nocturnal asthma group), and 72 healthy people (control group). All people investigated were of Chinese Han nationality. RESULTS: The distribution frequency of genotype Arg/Arg, Arg/Gly, and Gly/Gly at beta(2)-AR 16 loci was 12, 16 and 72% in the nocturnal asthma group; and 27, 41 and 32% in the non-nocturnal asthma group. There was a significant increase in the frequency of genotype Gly/Gly and allele Gly in the nocturnal asthma group compared with the non-nocturnal asthma group (p < 0.01). However, there was no significant difference in the frequency of genotype Gly/Gly and allele Gly in the non-nocturnal asthma group, compared with the control group. There was no significance in the frequency of the genotypes and alleles of beta(2)-AR 27 loci among the three groups (p > 0.05). CONCLUSION: The Gly16 polymorphism of beta(2)-AR was overrepresented in nocturnal asthmatic patients, correlated with nocturnal asthma, and therefore appeared to be an important genetic factor in the expression of this asthmatic phenotype. PMID- 16369121 TI - Helicobacter pylori seroprevalence in patients with chronic obstructive pulmonary disease and its relation to pulmonary function tests. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a slowly progressive condition characterized by poorly reversible airflow limitation that is associated with an abnormal inflammatory response of the lung. It has been shown that there is a seroepidemiological association of Helicobacter pylori (Hp) infection with many inflammatory conditions. OBJECTIVE: In this study, we aimed to investigate seroprevalence in Hp patients with COPD and to determine whether there is an association between Hp infection and COPD. METHODS: Forty-nine voluntary patients with COPD and 50 healthy control subjects of similar age and sex were included in the study. Hp-specific IgG was measured with a commercially available kit from venous blood samples. RESULTS: Serum levels of Hp-specific IgG and Hp IgG seropositivity were significantly higher in the patients with COPD than in the control subjects (p < 0.001 and p = 0.006, respectively). In addition, when the patients with COPD were grouped according to Hp IgG seropositivity, forced expiratory volume in 1 s (FEV(1)) values were lower in the seropositive patients compared to seronegative patients, and Hp serum IgG levels were correlated with FEV(1) values, which indicate the severity of COPD, in the COPD group (r = -0.306, p = 0.032). CONCLUSION: The results suggest that there is an association between Hp infection and COPD, and Hp IgG levels are correlated with the severity of COPD. PMID- 16369122 TI - Natural course of isolated pulmonary langerhans' cell histiocytosis in a toddler. 3-year follow-up. AB - Isolated pulmonary Langerhans' cell histiocytosis (LCH) is distinctly rare under the age of 15 years, since the majority of patients are young adult males with heavy smoking habits. Isolated pulmonary involvement suggests that antigens inhaled from cigarette smoke are involved. Here we present a case of LCH restricted to the lungs in a toddler whose parents were heavy smokers. Since LCH was not medically treated for 3 years due to parental refusal, the disease can be regarded as having followed its natural course. During the 3-year follow-up, the disease progressed to severe pulmonary fibrosis resulting in honeycomb lungs. Based on the comparative immunohistochemical analyses of the cells obtained from bronchoalveolar lavages during the disease course, it appears that the evolution of fibrosis is rather a result from the accumulating alveolar macrophages than from the persistence of the Langerhans' cells. Passive cigarette smoking may be considered a significant risk factor in both the pathogenesis and development of pulmonary LCH in a small child. PMID- 16369123 TI - Does cold environment affect Streptococcus pneumoniae adherence to rat buccal epithelium? AB - BACKGROUND: Colonization of the nasopharyngeal flora by Streptococcus pneumoniae increases in winter. However, in vitro studies suggest that bacterial adherence decreases in low temperatures. These contrasts may be related to the difference between in vitro and physiological conditions. OBJECTIVES: We investigated whether pneumococcal adherence to buccal epithelial cells of rats increases in cold temperature. METHODS: The adherence of S. pneumonia (ATCC 49619) was studied in six groups of rats. Basal pneumococcal adherence was checked after 1 h both at room temperature and in a cold environment (at 10 degrees C) for the evaluation of changes in basal flora in the first two groups (basal control). In the remaining four groups, the effect of cold treatment before or after inoculation of S. pneumoniae, or cold treatment during the whole study period was investigated. RESULTS: The adherence of inoculated pneumococci was found to be significantly decreased in rats exposed to cold air during the whole study period (group 3). However, the decrease was not significant in rats exposed to cold air after bacterial inoculation (group 6). In contrast, pneumococcal adherence was found to be significantly increased in rats staying at 24 degrees C room temperature (control, group 5), during the whole pre- and postincubation period. A slight increase was also detected in rats pretreated in cold air and then kept at room temperature (group 4) (p > 0.05). CONCLUSION: In contrast to our expectations, pneumococcal adherence was found to be decreased in cold environment in vivo. PMID- 16369124 TI - Elevated levels of antimicrobial peptides in bronchoalveolar lavage fluid in patients with chronic eosinophilic pneumonia. AB - BACKGROUND: Chronic eosinophilic pneumonia (CEP) is an idiopathic pulmonary disease. As the lung is in direct communication with the environment, inhaled antigen may activate immune mechanisms in the airway that may participate in the pathogenesis of idiopathic pulmonary diseases. Defensins are antimicrobial peptides that consist of alpha-defensin (HAD) in neutrophils and beta-defensin (HBD) in epithelial cells. Defensins act as innate immunity against pathogens acquired from the environment and as mediators to induce local inflammation. OBJECTIVES: The aim of the present study was to determine whether immune mechanisms in the airway are induced in CEP patients. METHODS: We measured BALF defensin levels in patients with CEP, acute EP (AEP) and drug-induced eosinophilic pneumonia (drug-EP). We also measured BALF levels of IL-5, GM-CSF, eotaxin and RANTES. These substances can recruit eosinophils. RESULTS: BALF HAD levels were higher in patients with CEP than in those with drug-EP and normal controls. HBD-2 was detected in BALF of 10 of 11 CEP patients and in 3 of 5 AEP patients while its level was below detection in drug-EP patients and normal controls. BALF HBD-2 levels correlated with the proportion of lymphocytes in CEP patients. CONCLUSION: The defensin-linked immune system is activated in CEP but not in drug-EP. This suggests that inhaled antigen(s) may be involved in the pathogenesis of CEP. PMID- 16369125 TI - Radiological findings, pulmonary function and dyspnea in underground coal miners. AB - BACKGROUND: Respiratory disability induced by dust exposure in coal workers is assessed by pulmonary function tests and radiological evidence of pneumoconiosis. High-resolution computed tomography (HR-CT) improves the visibility of tissue changes, but the value of the findings for the clinical evaluation is controversial. OBJECTIVES: It was the aim of this study to evaluate the correlation between the International Labour Office (ILO) classification and the degree of emphysema in HR-CT with self-reported dyspnea and pulmonary function tests including diffusion capacity for CO (DL,CO). METHODS: We investigated 87 coal miners (aged 67+/-6 years), having worked underground for 26+/-9 years, with pulmonary function tests and HR-CT. Univariate associations were tested with correlation coefficients, and multivariable analyses used a stepwise forward regression model. RESULTS: No aspect of the ILO classification showed a univariate correlation with dyspnea or forced expiratory flow in 1 s (FEV1). Emphysema CT score was strongly associated with DL,CO (rs=-0.40; p<0.001) and FEV1/maximal vital capacity (r=-0.38; p<0.001) in univariate analysis, but not with the clinical grade of dyspnea (r=-0.14; p=0.256). CT emphysema score but not ILO classification was associated with FEV1 in multivariable analyses (rs=-0.37; p<0.001). Dyspnea was best approximated by DL,CO (r=-0.312; p=0.008). CONCLUSION: The clinical grade of breathlessness was best approximated by DL,CO. HR-CT showed a good association with expiratory flow limitation. ILO classification of the chest radiograph may be a marker of exposure but conveys little information about the degree of respiratory impairment. PMID- 16369127 TI - Infliximab treatment in a patient with systemic sclerosis associated with lung fibrosis and pulmonary hypertension. AB - This is the first report of the efficacy of anti-TNFalpha treatment in a patient with lung fibrosis and pulmonary hypertension associated with advanced systemic sclerosis, refractory to conventional therapies. The patient was treated with infliximab (5 mg/kg) and methotrexate (10 mg/week) for 1 year. After 6 months of therapy, the echocardiogram showed a reduction in pulmonary pressure, confirmed after 1 year. During treatment, the patient's quality of life improved significantly and high-resolution computed tomography of the chest, lung function tests and blood gas analysis remained stable. After 1 year, the patient decided to stop infliximab therapy (for family reasons related to the distance to our hospital). Lung function tests, pulmonary arterial pressures and blood gas analysis progressively worsened and the patient died 11 months later. Few open label studies have been conducted on the efficacy of anti-TNFalpha therapy in patients with systemic sclerosis. Here we report our experience in a case of systemic sclerosis complicated by pulmonary fibrosis and hypertension. Infliximab treatment seemed effective, suggesting that controlled randomized trials to evaluate infliximab efficacy in these patients and to compare infliximab with other anti-TNFalpha treatments would be worthwhile. PMID- 16369128 TI - Interstitial pulmonary amyloidosis. PMID- 16369131 TI - Metabolic syndrome. A new controversy. PMID- 16369129 TI - Neutrophil apoptosis, activation and anti-inflammatory cytokine response in granulocyte colony-stimulating factor-treated patients with community-acquired pneumonia. AB - BACKGROUND: Despite antibiotic treatment, the mortality of severe community acquired pneumonia (CAP), especially in patients with severe comorbidity, remains high. Innate defense mechanisms including polymorphonuclear neutrophil (PMN) activation and survival, orchestrated by cytokines, are primarily responsible for the elimination of bacterial organisms from the alveolus. OBJECTIVES: The aim of this study was to evaluate the effect of granulocyte colony-stimulating factor (G CSF) on PMN activation, apoptosis and cytokine response in patients with CAP. METHODS: Patients received a single dose of G-CSF (1 x 300 or 480 microg s.c.) prior to standard antibiotic treatment (n=8) or standard treatment only (n=8). Apoptosis rate and expression of CD11b, CD66b, CD64 and CD114 surface molecules on systemic PMN were assessed using fluorescence-activated cell sorter analysis. Levels of the interleukin-1 receptor antagonist (IL-1 RA), the soluble tumor necrosis factor receptor inhibitor (sTNF-p55) and G-CSF were measured by ELISA. RESULTS: In the treatment group, 12 h after G-CSF application, neutrophil count increased, neutrophil activation marker CD11b was stimulated (CD11b: 48.6+/-9.7 vs. 71.2+/-17.7, p<0.01), neutrophil apoptosis decreased (apoptosis: 1.36+/-0.27 vs. 0.2+/-0.12%, p <.01) and the concentration of IL-1RA and sTNF-p55 increased (IL-1RA 136.4+/-72.2 vs. 340.1+/-194.6 ng/ml, p<0.01; sTNF-p55,382+/-4,243 vs. 632+/-4,714 ng/ml, p<0.01; control group nonsignificant). These effects were not seen in the control group. CONCLUSIONS: The application of a single dose of G-CSF in patients with CAP caused a prolonged survival and increased activation of neutrophils combined with a sustained release of anti-inflammatory cytokines. PMID- 16369132 TI - Infectious diseases. Some recent threats. PMID- 16369133 TI - Does using the bispectral index (BIS) during craniotomy affect the quality of recovery? AB - One of the aims of neuroanesthesia is to provide early postoperative recovery and neurologic examination in patients undergoing supratentorial surgery. Our aim was to investigate the role of using the bispectral index (BIS) in recovery from anesthesia and altering drug administration in patients undergoing craniotomy. Fifty American Society of Anesthesiologists' (ASA) physical status I-II patients undergoing craniotomy were included in the study. The patients were randomly divided into two groups, and all patients received standard induction drugs, and 0.8%-1.5% sevoflurane was used for maintenance of anesthesia. In the BIS-guided group, the concentration of sevoflurane was titrated to maintain BIS at 40-60. In the control group, the anesthesiologist was blind to BIS, and the concentration of sevoflurane was changed according to the patients' hemodynamic changes. The hemodynamic data, BIS values, and sevoflurane concentrations were recorded every 15 minutes. In addition, the BIS value was recorded by the primary anesthetist in the BIS-guided group and by another independent anesthetist in the control group. At the end of the study, recovery criteria and Aldrete recovery scores were recorded every 15 minutes. Neurologic assessments were performed when the Aldrete score was 9-10. BIS values were higher, and sevoflurane concentrations (P < 0.05) and total doses of fentanyl (P < 0.01) were lower, in the BIS-guided group. Times to first spontaneous breathing, eye opening, and extubation (P = 0.035, P = 0.001, and P = 0.0001, respectively) were significantly shorter in the BIS-guided group. Time to an Aldrete score of 9-10 and adequate neurologic assessment were similar between the groups. In conclusion, BIS monitoring by supratentorial craniotomy under general anesthesia reduced the maintenance anesthetic concentration and narcotic drug usage and lowered the recovery times from general anesthesia. PMID- 16369134 TI - Clinical features of fever associated with poor outcome in severe pediatric traumatic brain injury. AB - We describe the incidence and etiology of fever and the relationship between fever characteristics and outcome in children with severe traumatic brain injury (TBI). We conducted a retrospective study of children <14 years and with Glasgow Coma Scale (GCS) score of <9 admitted to a level I pediatric trauma center intensive care unit (PICU) between 1998 and 2003. We examined whether fever characteristics were associated with poor outcome (hospital discharge GCS score <13 and discharge disposition of either death or discharge to a skilled nursing facility). PICU length of stay (LOS) and hospital LOS were also examined. Data are presented as means and medians (SD), and P < 0.05 reflects significance. Ninety-three records were reviewed. Patients were 5.7 (SD 4.1) years old, 70% were male, and the average admission GCS score was 5. Mortality rate was 14%. Forty-eight (52%) patients had fever, and 23 (48%) of those patients had infectious fever. Each additional febrile episode was associated with a twofold higher risk of patients having a hospital discharge GCS score of <13 (odds ratio 2.4, 95% confidence interval 1.2-5.0) and having a 0.4-day longer PICU LOS (P < 0.001). Patients with infectious fever had a 0.9-day longer PICU LOS (P < 0.001). Patients with any fever in the PICU had an increased HLOS (0.9 days; P < 0.001). Our data suggest that in severe pediatric TBI, both fever and infection were common, and both were associated with longer LOS. Patients with higher fever burden had poor hospital discharge GCS score. PMID- 16369135 TI - The effects of 10 degrees reverse Trendelenburg position on subdural intracranial pressure and cerebral perfusion pressure in patients subjected to craniotomy for cerebral aneurysm. AB - The aim of the current study was to examine the effects of 10 degrees reverse Trendelenburg position (rTp) on subdural intracranial pressure (ICP), cerebral perfusion pressure (CPP), and dural tension. Additionally, the relationship between preoperative Hunt and Hess (H and H) grade and the subdural ICP in patients scheduled for cerebral aneurysm surgery was investigated. Twenty-eight consecutive patients with a cerebral aneurysm were subjected to craniotomy in propofol/fentanyl or propofol/remifentanil anesthesia. Subdural ICP was measured after opening of the bone flap and exposure of dura. After reference measurements of subdural ICP and mean arterial blood pressure (MABP), the measurements were repeated during 10 degrees rTp. No significant differences between the anesthetic groups were disclosed. During 10 degrees rTp, a significant decrease in MABP, ICP, and jugular bulb pressure was observed whereas CPP remained unchanged. In H and H 0 patients (unruptured aneurysm), the ICP decreased from 2.9 +/- 2.6 mmHg to 0.4 +/- 2.2 mmHg at 10 degrees rTp. In H and H I to II patients, the ICP decreased from 9.3 +/- 3.8 mmHg to 4.6 +/- 3.3 mmHg at 10 degrees rTp. A significant difference in the mean baseline subdural ICP and DeltaICP (change in ICP) was found between patients with unruptured aneurysm and patients with subarachnoid hemorrhage (H&H I and II). Furthermore, the relationship between the subdural ICP at neutral position and DeltaICP was significant. In patients without intracranial hypertension, 10 degrees rTp decreases subdural ICP and dural tension in patients with ruptured as well as patients with unruptured cerebral aneurysm; CPP is unchanged. PMID- 16369136 TI - Effect of graded hyperventilation on cerebral metabolism in a cisterna magna blood injection model of subarachnoid hemorrhage in rats. AB - In subarachnoid hemorrhage (SAH) with cerebrovascular instability, hyperventilation may induce a risk of inducing or aggravating cerebral ischemia. We measured cerebral blood flow (CBF) and cerebral metabolic rates of oxygen (CMRO2), glucose (CMRglc), and lactate (CMRlac) at different PaCO2 levels after experimental SAH in rats (injection of 0.07 mL of autologous blood into the cisterna magna). Four groups of Sprague-Dawley male rats were studied at predetermined PaCO2 levels: group A: normocapnia (5.01-5.66 kPa [38.0-42.0 mm Hg]); group B: slight hyperventilation (4.34-5.00 kPa [32.5-37.5 mm Hg]); group C: moderate hyperventilation (3.67-4.33 kPa [27.5-32.4 mm Hg]); group D: profound hyperventilation (3.00-3.66 kPa [22.5-27.4 mm Hg]). Each of the four groups included eight rats with SAH and eight sham-operated controls. CBF was determined by the intracarotid Xe method; CMRo2, CMRglc, and CMRlac were obtained by cerebral arteriovenous differences. In both SAH rats and controls, hyperventilation decreased CBF in proportion to the decrement in PaCO2 without affecting either CMRO2, CMRglc, or CMRlac. In groups C and D, CBF decreased by 20%-35%, but CMRs were maintained by a compensatory increase in oxygen extraction fraction (OEF). The results show that even profound hyperventilation in this model of SAH is associated with an adequate increase in OEF so that CMRs of oxygen, glucose, and lactate remain similar to levels observed in normocapnic conditions. PMID- 16369137 TI - The effect of hypertonic (3%) saline with and without furosemide on plasma osmolality, sodium concentration, and brain water content after closed head trauma in rats. AB - Adding furosemide (F) to mannitol causes a greater decrease of brain volume, intracranial pressure, and brain water content (BW) as compared with mannitol alone. We examined whether adding F to hypertonic saline (HS) causes less increase of BW early after closed head trauma (CHT) as compared with HS alone. With institutional approval, 125 rats underwent sham surgery or CHT and then immediately received no treatment, HS (1.2 g/kg, 3% solution), or HS + F (2 mg/kg). In groups 1-10 (n = 8/group), the percent BW content was determined at 30, 60, or 120 minutes. In groups 11-14 (n = 8/group), physiologic values were determined at 0, 30, 60, and 120 minutes. At 120 minutes, the increase of BW caused by CHT (sham = 78.9 +/- 0.6% and CHT = 81.5 +/- 2.2%, mean +/- SD) was prevented by HS + F (78.0 +/- 0.8%) but not by HS (80.7 +/- 2.2%). Both HS and HS + F similarly increased plasma osmolality and sodium concentration. Post-CHT hypotension and acidosis (30 and 60 minutes) and decrease of hemoglobin concentration (120 minutes) were less with HS + F than with HS. We conclude that adding F to HS decreases BW without causing more increase of osmolality and Na than that caused by HS alone. PMID- 16369138 TI - Mild hypothermia promotes pro-inflammatory cytokine production in monocytes. AB - Hypothermia is often associated with compromised host defenses and infection. Deteriorations of immune functions related to hypothermia have been investigated, but the involvement of cytokines in host defense mechanisms and in infection remains unclear. We have previously shown that mild hypothermia modifies cytokine production by peripheral blood mononuclear cells. In this study, the effects of hypothermia on the monocytic production of several cytokines and nitric oxide (NO) were determined. Monocytes obtained from 10 healthy humans were cultured with lipopolysaccharide (LPS) under hypothermic (33 degrees C) or normothermic (37 degrees C) conditions for 48 hours. We performed flow cytometric analysis for simultaneous measurement of interleukin (IL)-8, IL-1beta, IL-6, IL-10, IL-12p70, and tumor necrosis factor (TNF)-alpha in culture supernatants. NO production was quantified as accumulation of nitrite in the medium by a colorimetric assay. Compared with normothermia, mild hypothermia raised the levels of IL-1beta, IL-6, IL-12p70, and TNF-alpha produced by monocytes stimulated with LPS. On calculating the ratios of these elevated cytokines to IL-10, however, only IL-12p70/IL-10 and TNF-alpha/IL-10 ratios were significantly elevated under hypothermic conditions. In contrast, hypothermia did not affect NO production. This study demonstrates that mild hypothermia affects the balance of cytokines produced by monocytes, leading to a pro-inflammatory state. Specifically, monocytic IL-12 and TNF-alpha appear to be involved in the immune alterations observed in mild hypothermia. However, the clinical significance of these phenomena remains to be clarified. PMID- 16369139 TI - The evolution of cerebral ischemia in a rat model of complete unilateral carotid artery occlusion with severe hypotension as detected by diffusion-, T2-, and postcontrast T1-weighted magnetic resonance images. AB - Severe internal carotid artery stenosis or occlusion is considered to be one of the important causes of stroke. The authors created a complete unilateral carotid artery occlusion model in 15 Sprague-Dawley rats, induced severe hypotension for at least 36 minutes by exsanguination with the target mean arterial pressure being equal or less than 35 mmHg, and investigated the temporal and spatial evolution of cerebral ischemia by diffusion-, T2-, and postcontrast T1-weighted magnetic resonance images. Cerebral ischemia was detected in most regions of the right middle cerebral artery territory during exsanguination. There was no significant relationship between ischemic lesion volume detected on apparent diffusion coefficient (ADC) map (ADC lesion volume) and infarction volume found on histopathology. However, there was a linear relationship between the change in ADC lesion volume at blood reinfusion (after reinfusion minus before reinfusion) and the enlargement of the lesion volume during the postreinfusion period (Y = 0.4X + 161.7, P = 0.0066) and a significant logarithmic correlation between the volume of vasogenic edema found on postcontrast T1-weighted image at 1 hour of the postreinfusion period and the enlargement of the lesion volume during the postreinfusion period (Y = 62.1 x logX - 115.4, P = 0.022). In conclusion, although it may be difficult to predict the outcome of cerebral ischemia (infarction volume) from the lesion volume during exsanguination, the evolution of cerebral ischemia may be partly predicted by lesion volume changes seen on the ADC maps at the time of the blood reinfusion or by the severity of blood-brain barrier disruption at the early stage of the postreinfusion period. PMID- 16369140 TI - Anesthesia for minimally invasive cranial and spinal surgery. AB - The field of minimally invasive neurosurgery has evolved rapidly in its indications and applications over the last few years. New, less invasive techniques with low morbidity and virtually no mortality are replacing conventional neurosurgical procedures. Providing anesthesia for these procedures differs in many ways from conventional neurosurgical operations. Anesthesiologists are faced with the perioperative requirements and risks of newly developed procedures. This review calls attention to the anesthetic issues in various minimally invasive neurosurgical procedures for cranial and spinal indications. Among the procedures specifically discussed are endoscopic third ventriculostomy, endoscopic transsphenoidal hypophysectomy, endoscopic strip craniectomy, deep brain stimulation, video-assisted thorascopic surgery, vertebroplasty and kyphoplasty, cervical discectomy and foraminectomy, and laparoscopically assisted lumbar spine surgery. PMID- 16369141 TI - Disturbances of sodium in critically ill adult neurologic patients: a clinical review. AB - Disorders of sodium and water balance are common in critically ill adult neurologic patients. Normal aspects of sodium and water regulation are reviewed. The etiology of possible causes of sodium disturbance is discussed in both the general inpatient and the neurologic populations. Areas of importance are highlighted with regard to the differential diagnosis of sodium disturbance in neurologic patients, and management strategies are discussed. Specific discussions of the etiology, diagnosis, and management of cerebral salt wasting syndrome, the syndrome of inappropriate antidiuretic hormone secretion, and central diabetes insipidus are presented, as well as the problems of overtreatment. The importance of diagnosis at an early stage of these diseases is stressed, with a recommendation for conservative management of milder cases. PMID- 16369142 TI - Anesthesia for functional neurosurgery: review of complications. AB - The use of functional stereotactic neurosurgery is increasing for treatment of patients with movement disorders and other chronic illnesses. The anesthetic considerations include the influence of the anesthetic agents on the microelectrode recordings and stimulation testing of an awake patient. The purpose of this study was to review the anesthetic management and incidences of intraoperative complications during functional neurosurgery in our institution. One hundred seventy-eight patients underwent an ablative procedure (n = 6) or the insertion of deep brain stimulator (n = 172) under monitored anesthesia care for movement disorders (n = 124), chronic pain (n = 20), and other procedures (n = 34). Local anesthetic was used for head frame pin sites and burr holes. No sedation/analgesia was administered to 57 (32%) patients. One patient required conscious sedation and another general anesthesia for the entire procedure. The remainder received small increments (mean +/- SD) of propofol (113 +/- 73 mg), midazolam (1.6 +/- 0.8 mg), and/or fentanyl (93 +/- 55 mug). Intraoperative complications that occurred in 16% of the patients included seizures (n = 8), change in neurologic status (n = 5), airway obstruction (n = 2), and hypertension (n = 7). Functional neurosurgery can be performed with minimal anesthesia in many patients. Awareness and vigilance can improve the identification and early treatment of intraoperative complications such as seizures, loss of airway, and changes in the neurologic status. PMID- 16369143 TI - Possible role of the C-reactive protein and white blood cell count in the pathogenesis of cerebral vasospasm following aneurysmal subarachnoid hemorrhage. AB - The delayed ischemic neurologic deficit (DIND) is a common and potentially devastating complication in patients who have sustained subarachnoid hemorrhage (SAH). Recent evidence suggests that various constituents of the inflammatory response may be critical in the pathogenesis of this ischemic complication. The aim of this study was to evaluate the possible relationship between the C reactive protein (CRP)/white blood cell (WBC) count and DIND. A total of 88 patients with acute SAH were included. CRP and WBC count were estimated on a daily basis. Outcome was evaluated 1 year after the initial ictus according to the Glasgow Outcome Scale. CRP levels on days 5, 6, 7, and 8 were statistically significantly higher in the group of patients developing a DIND (P < 0.025, P < 0.016, P < 0.011, P < 0.0002). WBC counts were higher in this patient group on days 1, 4, 5, 6, and 7 (P < 0.0253, P < 0.0087, P < 0.00167, P < 0.0026, P < 0.0045). Overall CRP values were higher with increasing severity of the initial ictus according to the Hunt and Hess Scale and to the outcome according to the Glasgow Outcome Scale from day 3 on. A statistically significant relationship between WBCs and outcome could not be observed. The presented data do not prove that WBCs and CRP values have a direct contribution to the pathogenesis of ischemic complications following SAH, but it supports the assertion that inflammation may present a common pathogenic pathway in the development of such complications. PMID- 16369144 TI - Airway management in patients with pituitary disease: a review of 746 patients. AB - Difficulties in airway management are observed among patients with pituitary disease. The purpose of this study was to better characterize the relationship between disease factors such as tumor characteristics on difficult endotracheal intubation. The perioperative records of 746 patients that underwent transsphenoidal microsurgery at the University of Virginia between January 1995 and June 2001 were reviewed. Among the 746 patients studied, difficulty with endotracheal intubation was encountered in 28 patients (3.8%). Patient gender and tumor size were not associated with a difference in the incidence of unanticipated airway management difficulty. Unanticipated difficulty with airway management was more than three times more common in acromegalic patients (n = 121) than in patients with nonfunctioning pituitary tumors (9.1% [5.8-14%] vs 2.6% [1.5-4.5%], P = 0.007). Patients with Cushing disease (n = 182) and patients with a prolactinoma (n = 87) were no more difficult to intubate than patients with nonfunctioning tumors. Among patients with acromegaly, neither tumor size nor patient gender has any impact on the incidence of intubation difficulty. Among all patients, when difficulty was encountered, intubation assisted by the gum elastic bougie was successful 100% of the time. The incidence of difficulty in intubation is not higher among patients with pituitary disease than in the general surgical population; however, patients with acromegaly have difficult intubations three times more often. PMID- 16369145 TI - Desflurane improves the recovery of the evoked postsynaptic population spike from CA1 pyramidal cells after hypoxia in rat hippocampal slices. AB - Desflurane is a volatile anesthetic that allows rapid induction and emergence, reduces cerebral metabolism, and enhances tissue perfusion. We studied the effect of treatment with 4%, 6%, and 12% desflurane on hypoxic neuronal damage by comparing the size of the postsynaptic evoked population spike recorded from the cornu ammonis 1 (CA1) pyramidal cell layer of rat hippocampal slices before and 2 hours after a hypoxic insult. When the tissue was treated with 6% desflurane before, during, and after 3.5 minutes of hypoxia, recovery was significantly better in slices exposed to desflurane (37% +/- 9%) compared with untreated hypoxic slices (15% +/- 5%). A lower (4%) or higher (12%) concentration of desflurane did not significantly improve recovery after 3.5 minutes of hypoxia. In the period before hypoxia, 12% and 6% desflurane significantly increased the latency and decreased the amplitude of the postsynaptic population spike; 4% desflurane had a similar but nonsignificant effect on latency and amplitude. We conclude that 6% desflurane, a clinically useful concentration (1 minimal alveolar concentration), improved the recovery of postsynaptic evoked responses in rat hippocampal slices after 3.5 minutes of hypoxia. In vivo studies must be conducted to assess the potential clinical significance of 6% desflurane's neuroprotective activity. PMID- 16369147 TI - Pressure support mode improves ventilation in "asleep-awake-asleep" craniotomy. PMID- 16369148 TI - A ChemoSensory questionnaire for patients treated for cancer of the head and neck. AB - OBJECTIVES: To investigate primary chemosensory issues experienced by patients who undergo treatment for cancer of the head and neck and to develop and assess a ChemoSensory Questionnaire (CSQ). DESIGN: Cross sectional survey for questionnaire development and testing. METHODS: Literature, expert opinion, and focus group methodology were used to identify aspects of smell and taste affecting patients before, during, and after therapy for cancer of the head and neck. A draft instrument was compiled, reviewed, and revised. The revised instrument was administered along with a demographic and clinical form, the SF 12, the Performance Status Survey, and the University of Michigan Head and Neck Quality of Life Questionnaire (HRQOL) to a sample of patients. After item and scale analyses and reductions, a final instrument was assembled. Construct validity and test-retest reliability were assessed. RESULTS: Two hundred six patients were included in the study. The final survey instrument consisted of eight items, four on smell and four on taste. Cronbach's alpha was 0.89 for the smell scale and 0.78 for the taste scale. Correlations with other HRQOL scales ranged from 0.20 to 0.64 for the taste scale and from 0.10 to 0.33 for the smell scale. Good construct validity of the CSQ scores was demonstrated. CONCLUSIONS: Treatment for head and neck cancer negatively impacts smell and taste. A survey instrument was developed to evaluate chemosensory function that is simple to administer and is brief. This instrument will be of value in identifying factors that contribute to chemosensory disturbance and may serve as a guide for planning treatment regimens that minimize such disturbance. PMID- 16369149 TI - Aerodynamics of the human larynx during vocal fold vibration. AB - OBJECTIVES: : The goal of this study was to comprehensively analyze the influence of aerodynamics on laryngeal function. Three specific aspects were considered: 1) a multidimensional comparison of the interaction of subglottic pressure, sound intensity, and fundamental frequency; 2) examination of instantaneous changes in subglottic pressure during each glottic cycle; and 3) determination of the threshold subglottic pressure for vocal fold vibration and its dependence on other aerodynamic factors. STUDY DESIGN: : Prospective study with six healthy individuals without history of voice disorders. METHODS: : The subjects vocalized the vowel sound /i/ with a variety of different intensities, pitch, and sound intensity. Subglottic air pressure, fundamental frequency, sound intensity, and the electroglottography signal were simultaneously measured. RESULTS: : Linear relationships were seen in all subjects between subglottic air pressure and sound intensity, although there were large variations in the slopes of these relationships. Rapid variations in subglottic pressure during each glottic cycle were detected, corresponding to the opening of the vocal folds with each individual vibration. Threshold pressures for vocal fold vibration were dependent primarily on sound intensity and fundamental frequency and tended to be higher at vibration onset than at offset. CONCLUSION: : The larynx responded in a predictable pattern to general aerodynamic forces, but there was tremendous variability in its specific behavior. Fundamental frequency and sound intensity tended to increase with subglottic air pressure, but that relationship was not seen consistently in all subjects. The relationship between subglottic air pressure and sound intensity was usually linear, unlike the exponential relationship seen in previous studies. Subglottic pressure was noted to undergo rapid change with each glottic cycle in some, but not all, subjects and was most strongly affected by average subglottic pressure. Phonation threshold air pressure was influenced by the sound intensity and, to a smaller extent, by the fundamental frequency of the voiced sound. PMID- 16369150 TI - Urgent surgical airway intervention: a 3 year county hospital experience. AB - OBJECTIVES: To review the indications, complications, and outcomes of patients undergoing urgent surgical airway intervention. METHODS: The inpatient and outpatient charts of patients who underwent awake tracheostomy or were converted from cricothyrostomy to tracheostomy at our institutional-affiliated County Hospital over a 3 year period were reviewed. RESULTS: Ninety patients underwent awake tracheotomy, and seven were converted from cricothyrostomy to tracheostomy. Indications for awake tracheotomy included impending airway obstruction from malignancy of the aerodigestive tract or that causing extrinsic compression of the airway in 72 (80%) patients, neck abscess in 4 (4.4%), subglottic stenosis in 3 (3.3%), and a variety of other etiologies in 11 (12.2%). Thirty-eight (42%) patients were noted to have stridor. On fiberoptic laryngoscopy, 66 (80%) patients had moderate or severe airway obstruction, whereas 17 (20%) had mild or no obstruction. Of the 72 patients with squamous cell carcinoma, only 6 (8%) have been subsequently decannulated. Among the remainder, 13 of 18 (72%) have been decannulated. Complications occurred in seven (7.8%) patients after awake tracheotomy, none with untoward sequelae. Three severe complications occurred among the seven patients converted from cricothyrostomy to tracheostomy: anoxic brain injury in each, leading to death in two. CONCLUSIONS: Awake tracheostomy should be considered in any patient with impending or ongoing airway obstruction or with potential for difficult intubation. This should be performed in a timely manner before an emergent situation arises because the complications of emergency surgical airway can be devastating. PMID- 16369151 TI - Parathyroid hormone as a predictor of hypocalcemia after thyroidectomy. AB - OBJECTIVES: The aims of this study are to ascertain whether parathyroid hormone (PTH) assay before total/completion thyroidectomy followed by levels immediately thereafter in the recovery room and the morning after surgery are a reliable predictor of hypocalcemia at our institution. In addition, to determine the feasibility of early discharge home from hospital after total thyroidectomy on the basis of postoperative PTH levels. STUDY DESIGN: Prospective clinical study. METHODS: Two hundred one patients undergoing total thyroidectomy at our institution were enrolled in the study. Preoperative levels of PTH, serum calcium, ionized calcium, and albumin were drawn. In the postoperative phase, the same biochemical markers were taken in the recovery room within 1 hour of the procedure and at a standard time the morning after the procedure. Discharge planning took place as per existing standards. RESULTS: Two hundred and one patients were reviewed. PTH levels below 1.1 pmol/L within 1 hour of completion or total thyroidectomy were predictive of risk for developing hypocalcemia, requiring calcium replacement therapy. PTH levels above 1.6 pmol/L were predictive of patients who would remain normocalcemic. Patients who fell in between these two values require further monitoring with conventional calcium levels. CONCLUSIONS: At our institution, PTH assays drawn postoperatively are predictive of patients who are at risk for developing hypocalcemia. Early identification of at-risk patients will facilitate prompt calcium replacement therapy and potential safe early discharge from hospital. PMID- 16369152 TI - Does severity of obstructive sleep apnea/hypopnea syndrome predict uvulopalatopharyngoplasty outcome? AB - OBJECTIVES: Uvulopalatopharyngoplasty (UP3) is the single most commonly performed surgical procedure for the treatment of obstructive sleep apnea/hypopnea syndrome (OSAHS), but its success is limited. Our objective was to determine whether severity of disease of OSAHS based on polysomnography (PSG) data (apnea/hypopnea index [AHI]) is a significant factor in predicting successful treatment by UP3. In addition, we compared anatomic staging with severity of disease to determine which is the better predictor of success. STUDY DESIGN: A retrospective chart review of 134 patients who underwent UP3 as an isolated procedure for the treatment of OSAHS in a tertiary university-affiliated medical center. RESULTS: Forty-five patients had mild disease with an AHI less than 20. The surgical success rate (defined as a 50% reduction in AHI and a postoperative AHI of < 20) was 26.7%. There were 40 patients with moderate disease (AHI 20-40) who had a surgical success rate of 42.5%. There were 49 patients with severe disease (AHI < 40) with a surgical success rate of 26.5%. The same patients were analyzed by the Friedman Staging System using anatomic findings without incorporating the severity of disease. Results indicated the following. Stage I had a success rate of 80.6%, stage II patients had a success rate of 37.9%, and stage III had a success rate of 8.1%. Assessment of severity within each stage did not affect outcome. CONCLUSIONS: Patients with mild disease based on PSG data do not have a better chance of successful treatment than patients with severe disease. Severity of disease should not be incorporated in the staging system. The Friedman Staging System based on anatomic factors is superior to severity of disease as a predictor of successful UP3. PMID- 16369153 TI - Predictive factors for posterior triangle metastasis in HNSCC. AB - OBJECTIVE: Surgical modifications sparing uninvolved structures such as the spinal accessory nerve have been implemented since the advent of the radical neck dissection in 1906. The increased morbidity to the spinal accessory nerve involved with the dissection of level V lymph nodes has led to much controversy. In this study, we examine the incidence of nodal metastasis to all nodal levels involved with upper aerodigestive squamous cell carcinoma and attempt to determine when level V dissection is indicated. STUDY DESIGN: Retrospective chart review. METHODS: A study of all radical and modified radical neck dissections was performed at Louisiana State University - Shreveport Health Sciences Center and Overton Brooks Veterans Administration Hospital between 1996 and 2003 for upper aerodigestive squamous cell carcinoma. Univariate and multivariate analyses were performed to determine which neck and patient factors were significantly associated with level V metastasis. RESULTS: Seventy-nine patients with a total of 94 neck dissections were analyzed. The prevalence of level V metastasis was 7.4% of the total neck dissections. Multivariate analysis found that positive lymph nodes involving levels II, III, and IV was the only independent significant factor for level V metastasis (P = .0003). CONCLUSION: Our study is in concordance with other studies in the literature, revealing a low prevalence of level V metastasis in upper aerodigestive squamous cell carcinoma. Unlike other studies, we have found if levels II, III, and IV lymph nodes are found to be positive, dissection of level V is warranted. PMID- 16369154 TI - Objective computerized versus subjective analysis of facial synkinesis. AB - OBJECTIVES/HYPOTHESIS: To compare the sensitivity of an objective, computerized approach to measurement of facial synkinesis with that for a subjective approach and to examine the test-retest reliability of these approaches. STUDY DESIGN: Prospective, nonrandomized, and blinded. METHODS: Remote facial motion at the upper eyelids and oral commissures during a closed-lip smile and eyelid closure were measured using a commercially available computerized motion analysis system. Thirty healthy adults with normal facial nerve function were enrolled to establish normative data and a threshold value for synkinesis. Thirty consecutive patients were analyzed based on the synkinesis threshold value. Blinded subjective evaluations by two observers were also performed independently on the same patients. RESULTS: Facial synkinesis was detected significantly more frequently with objective than subjective analysis for eyelid closure; no significant differences between approaches were shown for the closed-lip smile. Interestingly, five (17%) patients developed early synkinesis within 3 months from the onset of the facial nerve injury. The test-retest reliability of all objective measures was excellent for both expressions. Close agreement in percent of patients with synkinesis between test and retest subjective evaluations was obtained for both observers for both expressions. CONCLUSIONS: The objective, computerized approach to detection of facial synkinesis has excellent reliability and is more sensitive than the subjective approach to assessment to the presence of facial synkinesis associated with eyelid closure. The finding of early synkinesis suggests that central nervous reorganization plays a role in regeneration of the facial nerve. PMID- 16369155 TI - The completely opacified frontal or sphenoid sinus: a marker of more severe disease in chronic rhinosinusitis? AB - OBJECTIVE: Determine whether complete opacification of the sphenoid or frontal sinus is associated with increased clinical severity of chronic rhinosinusitis (CRS). METHODS: Adult patients undergoing evaluation for CRS prospectively completed the rhinosinusitis symptom inventory (RSI) and underwent computed tomography of the paranasal sinuses. A cohort with at least one completely opacified frontal sinus was identified. To each patient in this opacified cohort, a control patient without complete frontal sinus opacification was matched with corresponding Lund score. Symptom scores for headache, facial pressure, RSI symptom domains, and medical resource consumption were statistically compared. Similar analysis was conducted for patients with complete sphenoid sinus opacification. RESULTS: Fifty-four patients with at least one completely opacified frontal sinus were matched for Lund score to the control group (mean Lund score 17.1). There was no statistically significant difference in headache (2.1 vs. 2.8) or pressure scores (2.0 vs. 2.3) or in the RSI symptom domains between those patients with completely opacified frontal sinuses and controls, respectively. Although completely opacified patients received more antibiotic treatment and missed more workdays, only the increased numbers of physicians visits (4.0 vs. 2.1, P = .02) was significant. Thirty-four of 35 completely opacified sphenoid patients were matched to control patients (mean Lund score 16.7). Again, differences in symptom scores for headache (2.5 vs. 2.5), facial pressure (2.3 vs. 2.3), and RSI symptom domains were not statistically significant. Increased use of antibiotics (9.6 vs. 3.9, P = .036) and physician visits (5.8 vs. 1.8, P = .024) for sphenoid opacification patients was statistically significant. CONCLUSIONS: Patients with a completely opacified sphenoid or frontal sinus do not necessarily manifest more severe clinical symptoms of CRS. Thus, a higher radiographic stage should not be automatically assigned to patients with a completely opacified sphenoid of frontal sinus in CRS. PMID- 16369156 TI - Development and validation of an objective instrument to measure surgical performance at tonsillectomy. AB - OBJECTIVES: The goals of this project were 1) to develop and validate an objective instrument to measure surgical performance at tonsillectomy, 2) to assess its interobserver and interobservation reliability and construct validity, and 3) to select those items with best reliability and most independent information to design a simplified form suitable for routine use in otolaryngology surgical evaluation. DESIGN: Prospective, observational data collection for an educational quality improvement project. METHODS: The evaluation instrument was based on previous instruments developed in general surgery with input from attending otolaryngologic surgeons and experts in medical education. It was pilot tested and subjected to iterative improvements. After the instrument was finalized, a total of 55 tonsillectomies were observed and scored during academic year 2002 to 2003: 45 cases by residents at different points during their rotation, 5 by fellows, and 5 by faculty. Results were assessed for interobserver reliability, interobservation reliability, and construct validity. Factor analysis was used to identify items with independent information. RESULTS: Interobserver and interobservation reliability was high. On technical items, faculty substantially outperformed fellows, who in turn outperformed residents (P < .0001 for both comparisons). On the "global" scale (overall assessment), residents improved an average of 1 full point (on a 5 point scale) during a 3 month rotation (P = .01). In the subscale of "patient care," results were less clear cut: fellows outperformed residents, who in turn outperformed faculty, but only the fellows to faculty comparison was statistically significant (P = .04), and residents did not clearly improve over time (P = .36). Factor analysis demonstrated that technical items and patient care items factor separately and thus represent separate skill domains in surgery. CONCLUSIONS: It is possible to objectively measure surgical skill at tonsillectomy with high reliability and good construct validity. Factor analysis demonstrated that patient care is a distinct domain in surgical skill. Although the interobserver reliability for some patient care items reached statistical significance, it was not high enough for "high stakes testing" purposes. Using reliability and factor analysis results, we propose a simplified instrument for use in evaluating trainees in otolaryngologic surgery. PMID- 16369157 TI - Investigation of C-arm cone-beam CT-guided surgery of the frontal recess. AB - OBJECTIVE/HYPOTHESIS: A cone-beam CT (CBCT) imaging system based on a mobile C arm (Siemens PowerMobil) incorporating a high-performance flat-panel detector (Varian PaxScan) has been developed in our laboratory. We hypothesize that intraoperative C-arm CBCT provides image quality and guidance performance sufficient to assist surgical approach to the frontal recess. STUDY DESIGN: A preclinical prospective study was conducted using six cadaver heads to assess the performance characteristics and the potential clinical utility of this imaging system. METHODS: The mobile C-arm was employed for intraoperative CBCT guidance of the endoscopic approach to twelve frontal recesses. RESULTS: The imaging system is capable of sub-mm 3D spatial resolution with bone and soft-tissue visibility and a field of view sufficient for guidance of head and neck surgery. The system can generate intraoperative, volumetric CT images rapidly with an acceptably low radiation exposure to the patient and with image quality sufficient for most surgical tasks. Moreover, the system is portable and compatible with the surgical setup, providing excellent access to the patient. Finally, the accuracy of the system is not bound to a registration process. CONCLUSIONS: The ability to create updated images as surgery progresses introduces the concept of 'near-real-time' CT guidance for head and neck surgery. We found that the use of CBCT increased surgical confidence in accessing the frontal recess, resolved ambiguities with anatomical variations, and provided valuable teaching information to surgeons in training in both preoperative planning and correlation between tri-planar CT scans and intraoperative endoscopic findings. PMID- 16369158 TI - Abnormalities of axon growth in human olfactory mucosa. AB - OBJECTIVES/HYPOTHESIS: Random biopsies of the human adult olfactory mucosa often demonstrate degenerative changes in the olfactory epithelium (OE) in both dysosmic and normosmic patients and, consequently, have limited diagnostic usefulness. However, detailed analysis of the subepithelial tissue with specific attention to the fascicles of the olfactory nerve and abnormalities of axonal growth may improve the correlation of histopathology with sensory function. STUDY DESIGN: Retrospective review of human OE biopsies. METHODS: Mucosal biopsies from the olfactory area obtained from 27 subjects were examined by light and electron microscopy, with particular attention to the olfactory nerve fascicles; results were correlated with clinical status. Immunohistochemical analysis was used to characterize the extent of axonal depletion, relative maturity of the parent population, and aberrant axonal growth. RESULTS: As expected, there are areas of respiratory metaplasia and neuronal depletion in normosmic as well as dysosmic patients. The degree of axon degeneration within the fascicles correlates better with individual olfactory status. Immature neurons predominate, and re-entrant neuromas develop in patients with olfactory loss caused by disconnection from the olfactory bulb. Individuals with olfactory loss caused by epithelial damage as with chronic rhinosinusitis display evidence of nerve fascicle degeneration and intraepithelial neuromas. CONCLUSION: The status of olfactory axons provides useful information on the overall condition of the olfactory periphery and improves the diagnostic usefulness of mucosal biopsies. In addition to an assessment of the epithelium per se, the fascicles of the olfactory nerve need to be characterized for a complete analysis of the olfactory mucosa. PMID- 16369159 TI - Implantation of the ossified cochlea: management with the split electrode array. AB - OBJECTIVES/HYPOTHESIS: To describe indications for, the surgical technique required, and the expected functional results of split electrode array cochlear implants. STUDY DESIGN: Retrospective chart review. METHODS: Data collected included etiology of deafness, radiographic findings, pre- and postoperative aided pure tone thresholds, and speech perception testing. Adult speech perception outcomes were measured using the Consonant Nucleus Consonant (CNC) monosyllable words and Hearing in Noise Test (HINT) in quiet/noise (+10 dB). The children were assessed using the Infants and Toddlers Meaningful Auditory Integration Scale. RESULTS: Five patients were implanted with a split electrode array. This included two adults and three children. Both adults had preoperative binaural aided pure tone averages worse than 50 dB and scores of 0% on both HINT quiet and CNC words. The children had undetectable preoperative aided thresholds and scored an average 4/40 on the IT-MAIS. Postimplant, the average threshold gain was 38.5 dB in the adults and 81.5 dB in the children. One adult improved to score 51%/22% on HINT quiet/noise at 6 months and 72%/30% at 12 months. The other adult continued to score 0% on HINT at 12 months but claimed substantial subjective auditory improvement after the first year of device use. The children averaged 28/40 on the IT MAIS at 6 months after implantation. Forty-two of 48 implanted electrodes were functional. CONCLUSIONS: The split electrode array is a useful alternative to traditional cochlear implants in treating deafened patients with cochlear ossification. Patients implanted with the split array show marked improvement in sound and speech perception. PMID- 16369160 TI - Retroauricular hairline incision for removal of upper neck masses. AB - BACKGROUND/OBJECTIVES: The general population has been increasingly concerned about cosmesis and hopeless about prominent or invisible postoperative scars of the face and neck. The purpose of this study was to evaluate the benefit of a retroauricular hairline incision (RAHI) by comparing it with conventional cervical incision for removal of upper neck masses. STUDY DESIGN: Prospective clinical study. METHODS: Thirty-four patients with upper neck masses were divided into two surgical groups of RAHI (17) and conventional cervical incision (17) matched by age, sex, marital status and size, location, pathology of lesions. The operation time, complications, length of hospital stay, and subjective satisfaction with incision scar checked by visual analogue scale were compared between groups. RESULTS: The lesions of each group were congenital cysts (6), abscesses (2), inflammatory masses (2), and benign (6) or malignant (1) tumors and located in the upper neck. Mean operation time was 51 +/- 17 minute in the RAHI group and 41 +/- 13 minutes in the controls (P = .064). Mean hospital stay and complication rates were comparable between groups. Mean score of patient's satisfaction was 8.9 +/- 0.7 in the RAHI group and 4.5 +/- 2.7 in the controls (P < .001). The degree of incision scarring did not differ between groups, but the scars were less visible in the RAHI group because of hiding by the auricle and hair. CONCLUSIONS: The postauricular approach leading to a potentially invisible area of operation has a clear cosmetic benefit compared with conventional cervical incision. This will be helpful for patients with surgically indicated upper neck masses who hope for invisible incision scars. PMID- 16369161 TI - The jaundiced gunn rat model of auditory neuropathy/dyssynchrony. AB - OBJECTIVE: High levels of bilirubin are neurotoxic and may result in deafness or auditory neuropathy/auditory dyssynchrony (AN/AD). The jaundiced (jj) Gunn rat animal model of kernicterus has electrophysiologic and neuroanatomic abnormalities of brainstem auditory nuclei with normal cochlear microphonic recordings. We examined morphologic changes in the cochlea, spiral ganglion, and auditory nerve and relate these findings to current understanding of AN/AD. METHODS: At 15 days of age, jj and nonjaundiced (Nj) littermates were injected with sulfadimethoxine (sulfa) and killed 3 days later by transcardial perfusion. Sections were cut through decalcified temporal bones, cochlear nerves, and auditory brainstem and processed for light and electron microscopy and immunohistochemical localization of calbindin-D and parvalbumin. RESULTS: Spiral ganglion neurons were severely degenerated with a paucity of myelinated axons in jj animals. Electron microscopy of the intramodilar auditory nerve revealed a lack of large caliber axons in jj-sulfa versus Nj-sulfa controls. Large diameter degenerating axons were characterized by an electron-dense atrophied axis cylinder resembling an axonopathy. CONCLUSIONS: Our findings of abnormal spiral ganglion cells and selective loss of large, myelinated auditory nerve fibers with no abnormalities in cochlear hair cells, support the sulfa-treated jj Gunn rat as a model for bilirubin induced AN/AD. The paucity of large caliber neurons undermines temporal coding of auditory information and neural synchrony and demonstrates that in addition to brainstem auditory nuclei, spiral ganglion neurons are selectively vulnerable to bilirubin toxicity. PMID- 16369162 TI - Spontaneous temporomandibular joint herniation into the external auditory canal: a case report and review of the literature. AB - OBJECTIVES: Case report of a man presenting with spontaneous temporomandibular joint (TMJ) herniation into the external auditory canal (EAC). STUDY DESIGN: Case report and literature review. METHODS: A case report of a patient with spontaneous TMJ herniation into the EAC is presented. RESULTS: The patient was a 41-year-old man complaining of tinnitus in his right ear. Examination revealed that the anterior wall of the bony EAC moved backward when the patient's mouth was closed and forward when opened, causing the resultant discomfort. In this case, surgery was performed, resulting in a good outcome with no complications. Thirteen other cases of spontaneous TMJ herniation into the EAC were found in the literature. CONCLUSION: Few cases of spontaneous TMJ herniation into the EAC have been reported; however, if surgical treatment is desired, we should comply with the patient's request. PMID- 16369163 TI - Confocal microscopy of the peripheral gustatory system: comparison between healthy subjects and patients suffering from taste disorders during radiochemotherapy. AB - OBJECTIVES: Laser-scanning microscopy (LSM) was used to compare taste buds and epithelia of fungiform papillae of healthy subjects with those of patients suffering from taste disorders during/after radiochemotherapy (RCT). Aim of the study was to investigate effects responsible for taste loss at a microscopic level. STUDY DESIGN: Prospective study. METHODS: Data from 12 healthy subjects (mean age 52.4, SD 9.5 years) were compared with those of 12 patients (mean age 54.7, SD 8.5 years) with head and neck cancer suffering from taste disorders during RCT. Four parameters from LSM were selected for analysis: 1) distance between the pore of the taste buds of fungiform papillae and the crest of the papillary vessels; 2) epithelial cells of each taste bud at 34 mum; 3), cell density, and 4) area of the taste pore at 4 mum. These data were correlated to measures of gustatory sensitivity obtained with both the validated "taste strips" test kit and electrogustometry. RESULTS: Patients complaining from taste disorders during RCT exhibited a significant decrease of taste function assessed with both natural and electric stimuli. In these patients, we found thicker epithelia and smaller areas of the taste pores compared with healthy subjects. In 30% of those patients, no taste pores were detectable; in deeper sections, however, normal taste buds were present. CONCLUSIONS: In conclusion, in RCT patients with taste disorders, LSM indicates changes of epithelia of fungiform papilla but no changes of the taste bud structure. Damage of the chorda tympani nerve by scattered rays, direct or indirect mucotoxic effects of chemotherapeutic agents, and covering of taste pores by epithelial cells are likely reasons for taste loss during RCT. PMID- 16369164 TI - Quality of life and head and neck cancer: a 5 year prospective study. AB - OBJECTIVES: Assessment of health-related quality of life (HRQL) in head and neck cancer patients from diagnosis to 5 years after start of treatment. STUDY DESIGN: A prospective, descriptive study METHODS: three hundred fifty-seven patients from Norway and Sweden filled in HRQL questionnaires, the European Organisation for Research and Treatment of Cancer (EORTC) QLQ-C30 and the EORTC QLQ-H and N35, six times during the first year and then after 5 years. At 5 years, 167 (87%) of the 192 living patients filled in questionnaires. RESULTS: Group data: clinical significant improvements in HRQL were not found between 1 and 5 years. Problems with teeth, opening of the mouth, dryness in the mouth, and sticky saliva were persistent or worsening. Similar findings were found regardless of sex, age, stage, or site when clinical significant changes are considered. Patients who died between 1 and 5 years reported reduced HRQL on 15 of 28 scales at 1 year compared with the survivors. Individual data: 40% of patients reported improved global HRQL from diagnosis to 5 years after start of treatment. In addition, 11% had "top scores" at both assessment points. The pattern in global HRQL scores also applies for most of the other function and symptom scales. CONCLUSION: After the first year after treatment, recovery of the persisting side effects of treatment cannot be expected for the group as a whole. Patients must be prepared for this. Some individual patients experience improvement in global HRQL. HRQL assessments in daily clinical practice can identify patients who are in need of additional support and symptom relief. PMID- 16369165 TI - Comparison of open dilatational tracheostomy with conventional pediatric tracheostomy in a growing animal model. AB - OBJECTIVES: To compare the tracheal changes after applying a new open dilatational tracheostomy (ODT) technique with those from a conventional open tracheostomy (COT) with vertical cartilage incision in a growing animal model. STUDY DESIGN: Prospective, experimental investigation in a rabbit model. MATERIALS AND METHODS: Thirteen New Zealand white rabbits as a pediatric model were divided into three groups: six rabbits had COT (n = 6), another six underwent an ODT (n = 6), and one rabbit acted as a control. Each rabbit underwent tracheostomy by assigned procedures on the first day. On day 8, they were decannulated. On day 15, their tracheas were harvested. We examined the gross findings and histologic changes of each tracheal segment at the stomal level. In addition, we analyzed three parameters: the quotient of the stomal and nonstomal segment in sagittal diameter, coronal diameter, and cross-sectional area. RESULTS: The framework of cartilages at the stomal level were more distorted in the COT group. Histologic examination also showed buckling of the anterior tracheal wall, loss of cartilage, infiltration by many polymorphonuclear neutrophils, and the marked ingrowth of fibrous tissue in the COT group. Sagittal and coronal diameters and cross-sectional areas were significantly affected more severely after a COT than after an ODT. CONCLUSION: Our new modification of percutaneous dilatational tracheostomy, named "open dilatational tracheostomy," was successfully applied to a small, growing animal model and showed more favorable and consistent healing of trachea compared with COT. Therefore, the authors' new tracheostomy procedure could be applied to children who require short-term tracheostomy at any age in clinical settings. PMID- 16369166 TI - Predictive factors and outcomes in endoscopic sinus surgery for chronic rhinosinusitis. AB - PURPOSE: To assess objective and quality of life (QOL) outcomes before and after endoscopic sinus surgery (ESS) in patients with chronic rhinosinusitis (CRS) and to determine preoperative factors that predict surgical outcome in these patients. METHODS: One hundred nineteen adult patients with CRS and a mean follow up of 1.4 +/- 0.35 years were evaluated prospectively including the following patient factors: prior sinus surgery, polyps, asthma, acetylsalicylic acid intolerance (ASA), smoking, allergy, depression, and sex. Computed tomography (CT), endoscopy, and QOL assessment was performed. Predictive value of patient factors was determined based on change in endoscopy and QOL scores after ESS. RESULTS: Objective outcomes: preoperative CT scores were significantly worse in patients with polyps, asthma, and ASA, whereas CT score was unaffected by prior sinus surgery, smoking, allergy, depression, and sex. Patients with CRS demonstrated significant improvement on nasal endoscopy after ESS, but preoperative, postoperative, and change in scores were affected by certain patient factors. Endoscopy scores were significantly worse in patients with prior sinus surgery, polyps, asthma, and ASA, but these patients also experienced the greatest improvement in endoscopy scores. Smokers and patients with depression had the least change in endoscopy scores. QOL outcomes: patients with CRS experienced improvement in QOL after ESS. Pre- and postoperative QOL was positively affected by polyps and adversely affected by ASA, depression, and female sex, but these groups still experienced significant improvement in QOL scores. Pre- and postoperative QOL was unaffected by prior sinus surgery, asthma, smoking, and allergies, and all of these groups experienced significant improvement in QOL scores. Factors predictive of outcome: ASA and depression were predictive of worse outcome. Preoperative CT scores approached significance as being predictive of outcome. CONCLUSION: Surgical management of CRS was associated with significant improvement on objective and QOL measures; however, specific patient factors, in particular ASA and depression, predict poorer outcome. Preoperative CT may be a predictor of endoscopic and QOL outcome and deserves further study. PMID- 16369167 TI - Positron emission tomography scan to determine the need for neck dissection after chemoradiation for head and neck cancer: timing is everything. AB - We present a case of a negative positron emission tomography (PET) scan in a patient with pathologic viable cancer at neck dissection. STUDY DESIGN: Case Report. METHODS: A 69-year-old man presented with clinical stage T2N2c squamous cell cancer of the left tonsil and was treated with definitive chemoradiation. Left-sided adenopathy decreased but remained palpable after therapy. RESULTS: PET scan performed 23 days after completion of treatment showed no suspicious uptake in the left neck. Neck dissection performed at 2 months post-therapy revealed viable tumor in left cervical nodes. CONCLUSIONS: Persistent adenopathy after chemoradiation for head and neck cancer remains a clinical dilemma. A negative PET scan is accurate but only if the scan is performed 3 to 4 months after therapy. PMID- 16369168 TI - Utility of a three-item smell identification test in detecting olfactory dysfunction. AB - OBJECTIVE: Physicians rarely assess smell function, largely because of time considerations. Therefore, there is clinical need for very brief cranial nerve I screening tests. Although a few such tests exist, none have been adequately validated. The goal of this study was to empirically assess the utility of a three-item microencapsulated odor identification test in detecting olfactory dysfunction. SETTING: Smell and taste center at a university medical center. METHODS: The test was administered to 224 consecutive patients (98 men and 126 women ranging in age from 15-88 years). As part of their overall assessment, the well-validated 40-item University of Pennsylvania Smell Identification Test (UPSIT) was also administered. Sensitivity, specificity, and both negative and positive predictive values of the three-item test were established relative to UPSIT dysfunction categories. Test-retest reliability was determined in a subset of 39 patients. RESULTS: The three-item test was abnormal in 99% (67/68) of patients with anosmia, 85% (35/41) of those with severe microsmia, 76% (31/41) of those with moderate microsmia, and 50% (17/34) of those with mild microsmia. Of the 40 normosmic patients, 62.5% (25/40) correctly identified all odors, 25% (10/40) two odors, and 12.5% (5/40) one odor. None of the normosmic patients missed all three items. Using a cut-off score of 2, the test's sensitivity and specificity were 99% and 40%, respectively, for detecting total anosmia. The corresponding negative and positive predictive values were 98% and 43%. For detecting anosmia and severe microsmia, these values were 93%, 45%, 88%, and 63%. For detecting any olfactory pathology, they were 82%, 63%, 42%, and 91%. The test retest reliability was 0.87. CONCLUSION: The brief three-item test used in this study was found to be highly sensitive in identifying olfactory loss in patients with chemosensory complaints, particularly those with severe dysfunction. Although only moderately specific, its high reliability and negative predictive value suggests it may be an appropriate screening test for olfactory loss. PMID- 16369169 TI - Upregulation of oncostatin m in allergic rhinitis. AB - OBJECTIVES: Oncostatin M is a multifunctional cytokine belonging to the interleukin-6 family of cytokines. It has been implicated as an important modulator of lower airway remodeling in the setting of asthma. However, there have been few studies regarding a similar role for the upper airway epithelium in the setting of allergic rhinitis. This study was undertaken to investigate the expression of oncostatin M mRNA and protein in normal and allergic rhinitis nasal mucosa and to localize the expression of the oncostatin M protein in allergic rhinitis. MATERIALS AND METHODS: Inferior turbinate mucosa samples from 20 patients with perennial allergic rhinitis and 20 matched normal control subjects were obtained. Oncostatin M mRNA was extracted from the inferior turbinate mucosae, then reverse transcriptase-polymerase chain reaction was performed and analyzed semiquantitatively. Differences in expression levels of oncostatin M protein between samples from allergic rhinitis patients and normal control subjects were analyzed through Western blot, and oncostatin M protein was localized immunohistochemically. RESULTS: The expression levels of oncostatin M mRNA and protein were significantly upregulated in patients with allergic rhinitis mucosa. Oncostatin M protein was predominantly localized in the surface epithelium, infiltrating inflammatory cells, vascular endothelium, and submucosal glands and was more strongly expressed in the nasal mucosa of patients with allergic rhinitis than in normal control subjects. CONCLUSIONS: Oncostatin M is expressed in the human nasal mucosa and is upregulated in the setting of allergic nasal inflammation. These results suggest a possible contribution of oncostatin M in the remodeling of the nasal mucosa in allergic rhinitis. PMID- 16369170 TI - Sentinel lymph node biopsy: a rational approach for staging T2N0 oral cancer. AB - OBJECTIVES/HYPOTHESIS: For oral cancer patients, the presence of neck nodal metastases is the most important disease prognosticator. However, a significant proportion of clinically N0 patients harbor occult microscopic nodal metastasis. Our objective was to determine the feasibility and accuracy of sentinel node biopsy (SNB) in the staging of T2N0 oral carcinoma patients. STUDY DESIGN: Prospective analysis. METHODS: Twenty patients with previously untreated N0 oral cavity squamous cell carcinoma were studied. Each patient had an SNB performed using preoperative technetium sulfur colloid lymphoscintigraphy, intraoperative gamma probe guidance, and intraoperative peritumoral injection of 1% isosulfan blue. All patients underwent neck dissection. The sentinel lymph nodes (SLNs) were sectioned in 2- to 3-mm intervals, formalin fixed, and sectioned at three levels. The non-SLNs were sectioned in a routine manner for histologic examination. RESULTS: SLNs were identified in all patients (100%) and accurately predicted the pathologic nodal status in 18 of 20 patients (90%). Tumor was found exclusively in the SLNs in six patients (30%). Two patients had positive SLNs at multiple neck levels. Two patients had a negative SLN and a positive non-SLN (false-negative findings). Occult nodal metastases were present in 60% of the cohort. CONCLUSIONS: SNB is a technically feasible and accurate procedure for staging the neck in oral carcinoma patients. However, SNB accuracy is lower for floor of the mouth lesions. The rate of occult nodal metastases identified in this cohort is higher than previously reported in the literature. These results suggest that SNB warrants further multi-institutional studies. PMID- 16369171 TI - DNA repair gene polymorphisms and risk of second primary neoplasms and mortality in oral cancer patients. AB - OBJECTIVES/HYPOTHESIS: We tested the hypothesis that polymorphisms in genes involved in DNA repair pathways are associated with the development of second primary neoplasms of the upper aerodigestive tract (UADT), as well as mortality, in patients previously diagnosed with oral squamous cell cancer (OSCC). METHODS: DNA specimens from 279 OSCC patients who had participated in two previous population-based case-control studies were assayed for the following polymorphisms: X-ray repair cross-complementing (XRCC) 1 Arg399Gln, XRCC3 Thr241Met, xeroderma pigmentosum complementing group D (XPD) Lys751Gln, and O methylguanine- DNA methyltransferase (MGMT) Leu84Phe and Val143Ile. Baseline demographic information was obtained from personal interviews and tumor characteristics and treatment were obtained from cancer registry files. Cox proportional hazards models were used to calculate hazards ratio (HR) estimates for each polymorphism in relation to the risk of developing second primary neoplasms at any site, UADT, and head and neck. HRs were also determined for associations with all-cause mortality and oral cancer specific mortality. RESULTS: A significant increased risk of second neoplasms (all sites combined, as well as for UADT sites and for head and neck squamous cell cancers) was observed among XRCC3 241Met allele homozygotes (HR 2.65-3.44, P < .02). No significant association with the development of second neoplasms was observed for the XRCC1 399Gln, XPD 751Gln, or MGMT 84Phe or 143Ile alleles. Although no associations with oral cancer-specific mortality were observed, we found a significant inverse association between all-cause mortality and possessing at least one copy of the XRCC1 399Gln allele (HR 0.68, 95% confidence interval [CI] 0.47-0.97, P = .03), as well as a suggestion of a direct association between all-cause mortality and having one copy of the XRCC3 241Met allele (HR 1.39, 95% CI 0.95-2.03, P = .09). CONCLUSIONS: Polymorphisms in the DNA repair enzyme gene XRCC3 241Met was associated with an increased risk of second neoplasms, and polymorphisms of the XRCC1 399Gln gene were associated with a decreased risk of all-cause mortality in patients with primary OSCC. These findings require confirmation in other populations before the clinical implications can be considered. PMID- 16369172 TI - Linking of bone morphogenetic protein-2 to resorbable fracture plates for enhancing bone healing. AB - OBJECTIVE: To test whether bone morphogenetic protein (BMP)-2 may be covalently linked to resorbable fracture repair plates using an ester-hydrolysis reaction and determining whether the linked compound can facilitate bone growth. STUDY DESIGN: Laboratory in vitro experiments. METHOD: Resorbable fracture repair plates were partially hydrolyzed using varying concentrations of acid or base. This intermediate was then reacted with EDAC (1-ethyl-3[-3-dimethylamino propyl carbodiimide) to form an EDAC intermediate, which was then reacted with either horseradish peroxidase (HRP), interleukin (IL)-2, or BMP-2. Compound binding to the plate was confirmed by immunofluorescent staining. Confirmation of protein function was determined by the following assays: HRP's ability to cleave peroxide, IL-2's ability to stimulate lymphocytes, and BMP-2's ability to stimulate C3H10T1/2 cells to generate alkaline phosphatase. RESULTS: Three compounds (HRP, IL-2, and BMP-2) were successfully linked to plates as confirmed by immunofluorescence staining or functional testing. Compounds demonstrated better covalent linking to plates under basic conditions. HRP, IL-2, and BMP-2 retained function after binding as measured by cleaved peroxide levels, lymphocytes proliferation, and alkaline phosphatase production. CONCLUSIONS: Covalent linking of compounds such as HRP, IL-2, and BMP-2 to resorbable plates is possible and represents a novel protein delivery technique. BMP-2 covalently linked to resorbable plates may be used to facilitate bone healing. Covalent linking of compounds to plates represents a novel method for delivering concentrated levels of growth factors to a specific site and potentially extending their half-life. Further investigation into this application for bone healing may lead to quicker healing. PMID- 16369173 TI - N-acetyltransferase 2 gene polymorphism and presbycusis. AB - OBJECTIVES/HYPOTHESIS: The enzyme of N-acetyltransferase (NAT) is involved in the metabolism and detoxification of cytotoxic and carcinogenic compounds as well as reactive oxygen species (ROS). The excessive amount of ROS generation occurs in the ageing inner ear. The exact etiopathogenesis of presbycusis is not known, but it is generally accepted that it is the result of series of insults, such as physiologic age-related degeneration, noise exposure, medical disorders and their treatment, as well as hereditary susceptibility. The effect of aging shows a wide interindividual range; we aimed to investigate whether profiles of NAT2 genotypes may be associated with the risk of presbycusis. STUDY DESIGN: Hospital-based, case-control study. METHODS: We examined 68 adults with presbycusis and 98 healthy controls. DNA was extracted from whole blood, and the polymorphisms of NAT2*5A, NAT2*6A, NAT2*7A/B, and NAT2*14A were determined using a real-time polymerase chain reaction and fluorescence resonance energy transfer with a Light Cycler Instrument. Associations between specific genotypes and the development of presbycusis were examined by use of logistic regression analyses to calculate odds ratios and 95% confidence intervals. RESULTS: Gene polymorphisms at NAT2*5A, NAT2*7A/B, and NAT2*14A in subjects with presbycusis were not significantly different from in the controls (P > .05). However, in NAT2*6A, the risk of presbycusis was 15.2-fold more in individuals with mutant allele than subjects with wild genotype (P = .013). Individuals with NAT2*6A heterozygote allele had a 0.34-fold less risk in the development of presbycusis than subjects with mutant allele (P = .032) CONCLUSION: We demonstrated a significant association between the NAT2*6A polymorphism and age-related hearing loss in this population. However, the sample size was relatively small, and further studies need to investigate the exact role of NAT2 gene polymorphism in the etiopathogenesis of the presbycusis. PMID- 16369174 TI - Sinonasal leiomyosarcoma: review of literature and case report. AB - OBJECTIVES/HYPOTHESIS: To determine prognosis of primary sinonasal leiomyosarcomas after treatment. STUDY DESIGN: Literature review and case report. METHODS: Review of English literature from MEDLINE and independent sources with the addition of our case. RESULTS: Including our case, 63 cases have been reported. Primary treatment includes resection with or without radiation. Chemotherapy has not been reported to be effective. In our case, however, chemotherapy, consisting of etoposide and high-dose ifosfamide, caused the tumor to shrink significantly. On the basis of a review of all reported cases, the overall survival rate at a mean follow-up of 38.24 month is 66%. The minimal overall survival rates at 5 and 10 years are 20% and 6%, respectively. CONCLUSION: The prognosis for primary sinonasal leiomyosarcomas is poor. However, a 10-year survival has been reported in a few patients. Chemotherapy may be a useful adjunct when managing extensive lesions unamenable to curative resection. PMID- 16369175 TI - Antivascular therapy of oral tongue squamous cell carcinoma with PTK787. AB - OBJECTIVES/HYPOTHESIS: Vascular endothelial growth factor (VEGF) is an important mediator in tumor vascularization, growth, and metastasis. We investigated whether blockade of the VEGF receptor (VEGF-R) signaling pathway by the tyrosine kinase inhibitor PTK787 combined with CPT-11, a semisynthetic camptothecin analogue, can inhibit the tumor growth and angiogenesis of squamous cell carcinoma of the oral tongue in an orthotopic nude mouse model. METHODS: JMAR human oral squamous cell carcinoma cells were injected into the tongues of nude mice. Seven days later, the mice were randomized to receive a placebo, daily oral PTK787, weekly CPT-11 injection, or PTK787 plus CPT-11. After 4 weeks of treatment, the mice underwent necropsy, and the tongue tumors, cervical lymph nodes, and lungs were removed for immunohistochemical analyses. RESULTS: CPT-11, PTK787, and PTK787 plus CPT-11 significantly decreased tumor volumes and prolonged survival. The combination treatment group had the most significant decrease in volume and increase in survival. PTK787 alone or in combination with CPT-11 reduced the phosphorylation of VEGF-R in tumor cells and tumor-associated endothelial cells, was associated with decreased microvessel density, a decreased proliferative index, and an increased apoptotic index. PTK787 alone or the combination therapy resulted in apoptosis of both tumor cells and tumor associated endothelial cells. CONCLUSIONS: These results suggest that targeting VEGF-R tyrosine kinase activity can be an effective therapeutic approach in squamous cell carcinoma of the oral tongue. PMID- 16369176 TI - Prevalence of laryngeal irritation signs associated with reflux in asymptomatic volunteers: impact of endoscopic technique (rigid vs. flexible laryngoscope). AB - OBJECTIVES: The objectives of this study were to 1) determine the prevalence of ENT findings in the normal asymptomatic population and 2) to compare findings between flexible and rigid laryngoscopes in an attempt to increase specificity of diagnosis of reflux in endoscopic laryngeal examinations. STUDY DESIGN: Prospective study. METHODS: Fifty-two nonsmoker volunteers (24 male, 28 female), mean age of 42.7 years, with no history of ENT abnormalities or gastroesophageal reflux disease, underwent both rigid and flexible videolaryngologic examinations with a digital endoscopic unit. A group of three expert judges reviewed the oral and transnasal examinations blindly and independently for physical signs of irritation/inflammation commonly associated with reflux. RESULTS: Atleast one sign of tissue irritation was detected in 93% and 83% of the population when using a flexible and a rigid laryngoscope, respectively. Results showed a high incidence of posterior commissure bar (53.2% and 51.9%), arytenoid complex edema/erythema (76.3% and 53.2%), and pseudosulcus (37.2% and 7.7%). Most signs were more frequently detected on flexible transasal examinations than with rigid transoral examinations: posterior pharyngeal wall (<0.01), interarytenoid irritation (<0.01), arytenoids complex irritation (<0.01), ventricular obliteration (<0.01), and pseudosulcus (<0.01). CONCLUSIONS: Several signs of posterior laryngeal irritation (e.g., interarytenoid bar, erythema of the medial wall of the arytenoids), which are generally considered to be signs of laryngopharyngeal reflux, are present in a high percentage of nonsymptomatic individuals, raising question about their diagnostic specificity. In addition, these signs were more often detected with flexible than with rigid laryngoscopes, suggesting that flexible laryngoscopy is more sensitive but less specific in identifying laryngeal tissue irritation. PMID- 16369177 TI - Transnasal, endoscopic vocal fold augmentation. AB - The practice of injection laryngoplasty under local anesthesia has become more common as both the indications for the procedure and the number of injectable substances increased. Modifications to the injection techniques used for vocal fold augmentation have been described over the last decade that reflect changes in the established percutaneous and transoral approaches. These percutaneous and transoral injection techniques for the treatment of dysphonia secondary to glottic incompetence are well described and provide an adequate approach for most cases. However, these traditional methods may be difficult to master, require great patient tolerance, and may be impossible to perform when anatomic or physiologic barriers exist. We describe a new application of the fiberoptic transnasal endoscope to perform laryngeal injection using a flexible needle through a port in the endoscope. This technique is easily mastered and readily tolerated by patients who would not be candidates for the other injection techniques under local anesthesia. We present our favorable experience with this technique and identify its shortcomings coupled with recommendations to address future technical modifications. PMID- 16369179 TI - Hmgb-1 as a therapeutic target for infectious and inflammatory disorders. AB - High-mobility group box (HMGB)-1 was recently identified as a lethal mediator of severe sepsis and represents a novel group of intracellular proteins that function as inflammatory cytokines when released into the extracellular milieu. From a clinical perspective, extracellular HMGB-1 can cause multiple organ failure and contribute to the pathogenesis of diverse disorders including sepsis, cardiovascular shock, rheumatoid arthritis, diabetes, and cancer. HMGB-1 has been proven to be a successful therapeutic target in experimental models of diverse infectious and inflammatory diseases, and these findings have renewed the clinical interest of specific cytokine inhibitors. However, little is known about the molecular mechanisms underlying the cytokine activity of HMGB-1 and its contribution to infection and inflammation. This article analyzes the value of HMGB-1 as a therapeutic target for the treatment of diverse infectious and inflammatory disorders and its interest for human clinical trials. PMID- 16369180 TI - Bone marrow failure in male rats following trauma/hemorrhagic shock (T/HS) is mediated by mesenteric lymph and modulated by castration. AB - Bone marrow (BM) suppression occurs following trauma/hemorrhagic shock (T/HS) in experimental animals as well as following severe injury in humans. Although the pathophysiology of BM suppression remains poorly understood, mesenteric lymph is thought to play an important role in T/HS-induced BM suppression; however, the direct effect of mesenteric lymph on BM in vitro has never been studied. In addition, recent studies in rats have also shown that female and castrated male rats are protected against T/HS-induced BM failure. We therefore hypothesized that mesenteric lymph is a source of factor(s) causing direct BM suppression and that the effects of mesenteric lymph are gender dependent. To test this hypothesis, we subjected noncastrated (NC) and castrated (C) male and proestrus female rats to T/HS or trauma sham shock (T/SS). Mesenteric lymph collected 3 h postshock was plated (4% v/v) with BM cells collected from unmanipulated male or female rats for granulocyte-macrophage colony-forming units (CFU-GM) and erythroid burst-forming units (BFU-E) colony growth. The T/HS lymph collected from NC-male rats but not from female rats caused a 50% inhibition of CFU-GM and BFU-E colony growth compared with cells cultured without lymph (P < 0.05 versus all other groups (ANOVA + Tukey). T/HS lymph collected from C-male rats also caused no significant inhibition of CFU-GM and BFU-E colony growth compared with cells cultured without lymph. Female and male BM progenitor cells had a similar response to mesenteric lymph from all groups tested. These results show that mesenteric lymph from NC-male rats suppresses CFU-GM and BFU-E progenitor growth in vitro, whereas the lymph from C-male and female rats did not. The effects of mesenteric lymph were the same regardless of whether the target BM was from male or female rats. The results therefore indicate that BM failure in male rats is directly mediated by factors present within the mesenteric lymph that appear to be modulated by castration, and protection against BM failure in female rats occurs at a systemic rather than a local level. Further studies are needed to elucidate potential therapeutic effects of lymph manipulation in hematopoiesis after injury. PMID- 16369181 TI - ROLE of endogenous peroxisome proliferator-activated receptor-alpha (PPAR-alpha) ligands in the development of gut ischemia and reperfusion in mice. AB - The peroxisome proliferator-activated receptor-alpha (PPAR-alpha) is a member of the nuclear receptor superfamily of ligand-dependent transcription factors related to retinoid, steroid, and thyroid hormone receptors. The aim of the present study was to examine the effects of endogenous PPAR-alpha ligand on the development of gut ischemia and reperfusion injury. Splanchnic artery occlusion (SAO) shock was induced in mice by clamping both the superior mesenteric artery and the celiac artery for 30 min, followed thereafter by release of the clamp (reperfusion). At 60 min after reperfusion, animals were sacrificed for histological examination and biochemical studies. SAO-shocked WT mice developed a significant increase of ileum tissue, TNF-alpha, IL-1beta, myeloperoxidase activity, and marked histological injury. SAO shock was also associated with a significant mortality (0% survival at 24 h after reperfusion). Reperfused ileum tissue sections from SAO-shocked WT mice showed positive staining for P-selectin, ICAM-1, TNF-alpha, and IL-1beta. Absence of a functional PPAR-alpha gene in PPAR alphaKO mice resulted in a significant augmentation of all the above-described parameters. Thus, endogenous PPAR-alpha ligands reduce the degree of ileum injury caused by ischemia and reperfusion. PMID- 16369182 TI - Efficiency of tazobactam/piperacillin in lethal peritonitis is enhanced after preconditioning of rats with O3/O2-pneumoperitoneum. AB - Insufflation of ozonized oxygen into the peritoneum (O3/O2-pneumoperitoneum [O3/O2-PP]) of rats reduced the lethality of peritonitis. We evaluated the prophylactic effect of O3/O2-PP combined with tazobactam/piperacillin (TZP) in polymicrobial lethal peritonitis. Wistar rats were conditioned by daily repeated insufflation of ozone for 5 days, and hematologic parameters were determined. Sepsis was induced by i.p. injection of cecal material derived from donor rats. Simultaneously, TZP was applied at a single dosage of 65 mg/kg or at two dosage schedules of 65 mg/kg each at an interval of 1 h. The conditioning effect of O3/O2-PP on the number of blood cells was measured before inoculation of bacteria. The mRNA levels of proinflammatory cytokine IL-lbeta and TNF-alpha were determined at 4 h post infection in spleen and liver by semiquantitative in situ hybridization analysis. Preconditioning of rats by O3/O2-PP enhanced the number of blood leukocytes and granulocytes and increased the survival rate of septic rats up to 33%. The combination of O3/O2-PP and TZP further enhanced the survival rate up to 93%. This effect was accompanied by a reduced amount of IL-1beta and TNF-alpha mRNA in spleen and liver. In contrast, in non-infected animals the combination of O3/O2-PP and TZP enhanced IL-1beta and TNF-alpha mRNA in the spleen and IL-1beta mRNA in liver when compared with TZP- and sham-treated controls. The preconditioning effect of O3/O2-PP seems to support the biological effectiveness of TZP by altering the immune status before and during the onset of sepsis. The combined therapy could be a simple, preoperative intervention for abdominal surgery to reduce postoperative morbidity and mortality. PMID- 16369183 TI - N-acetylcysteine protects the rats against phrenic nerve dysfunction in sepsis. AB - This study investigates the association of oxidative stress with the function of the phrenic nerve and inquires whether N-acetylcysteine (NAC) may counteract the possible detrimental effects. Thirty rats were divided into three groups: sham, cecal ligation and puncture (CLP), and CLP plus NAC treatment. Sepsis was produced by the CLP procedure. NAC was administered at 70 mg/day for 7 days. Electrophysiology was evaluated by the needle electromyography of the diaphragm and phrenic nerve conduction study. Oxidative stress was evaluated by malondialdehyde (MDA), nitrite/nitrate (NN), and reduced-glutathione (ReGSH) levels and myeloperoxidase (MPO) and catalase (CAT) activities in the phrenic nerve. In the CLP group, ReGSH and CAT were decreased (P = 0.0001, P = 0.07, respectively); and MDA, MPO, and NN were increased (P = 0.02, P = 0.0001, P = 0.043, respectively), compared with the sham group. NAC administration increased the ReGSH (P = 0.036) and decreased the MDA, MPO, and NN (P = 0.008, P = 0.01, P = 0.032, respectively), compared with the CLP group. In the CLP group, electrophysiology revealed reductions in the number of motor unit action potentials (P = 0.0001) and prolongations in the latency of the compound nerve action potential (P = 0.0001), indicating phrenic nerve neuropathy. NAC administration significantly ameliorated these electrophysiological alterations (P = 0.011, P = 0.0001, respectively), compared with the CLP group. The present results showed that intraabdominal sepsis is closely associated with phrenic nerve neuropathy. In addition, NAC administration protects the rats against the detrimental events of sepsis. PMID- 16369184 TI - Protective effect of a protein-bound polysaccharide, PSK, on CLP-induced sepsis in mice transplanted orthotopically with colon tumor. AB - We investigated the effects of a protein-bound polysaccharide, PSK, on the resistance of tumor-bearing mice against sepsis induced by cecal ligation and puncture (CLP). (a) In BALB/c mice that had received intracecal transplantation of colon 26 (C26) tumor, CLP with a 21-gauge needle significantly shortened the survival time, compared with that of non-tumor-bearing mice. Oral administration of PSK to such mice resulted in a significant prolongation of the survival time and increase of the survival rates. The effects were dependent on the timing of PSK administration and the dose. (b) CLP significantly increased the IL-10 level in serum, the IL-10 gene expression by spleen cells, the number of IL-10 producing CD4-positive T cells, and the productivity of IL-10 by spleen of tumor bearing mice compared with that of non-tumor-bearing mice. PSK administration to such mice suppressed the increase. Further, PSK prevented the reduction of gene expression of IFN-gamma and the number of IFN-gamma-producing CD4-positive T cells and IFN-gamma productivity by spleen cells of tumor-bearing CLP-treated mice. (c) Treatment with anti-IFN-gamma monoclonal antibody before CLP significantly reduced the effects of PSK. These findings suggest that the protective effect of PSK on the CLP-induced sepsis in mice transplanted orthotopically with C26 tumor is possibly mediated by suppression of IL-10 and promotion of IFN-gamma. PMID- 16369185 TI - Toll-like receptor 4, nitric oxide, and myocardial depression in endotoxemia. AB - The molecular mechanisms that mediate gram-negative sepsis-associated myocardial dysfunction remain elusive. Myocardial expression of inflammatory mediators is Toll-like receptor 4 (TLR4) dependent. However, it remains to be elucidated whether TLR4, expressed on cardiac myocytes, mediates impairment of cardiac contractility after lipopolysaccharide (LPS) application. Cardiac myocyte contractility, measured as sarcomere shortening of isolated cardiac myocytes from C3H/HeJ (with nonfunctional TLR4) and C3H/HeN (control), were recorded at stimulation frequencies between 0.5 and 10 Hz and after incubation with 1 and 10 mug/mL LPS for up to 8 h. Control cells treated with LPS were investigated with and without a competitive LPS inhibitor (E5564) and a specific inducible nitric oxide synthase (iNOS) inhibitor S-methylisothiourea. In control mice, LPS reduced sarcomere shortening amplitude and prolonged duration of relaxation, whereas sarcomere shortening of C3H/HeJ cells was insensitive to LPS. NFkappaB and iNOS were upregulated after LPS application in control mice compared with C3H/HeJ. Inhibition of TLR4 by E5564 as well as inhibition of iNOS prevented the influence of LPS on contractile activity in control myocytes. LPS-dependent suppression of cardiac myocyte contractility was significantly blunted in C3H/HeJ mice. Competitive inhibition of functional TLR4 with E5564 protects cardiac myocyte contractility against LPS. These findings suggest that TLR4, expressed on cardiac myocytes, contributes to sepsis-induced myocardial dysfunction. E5564, currently under investigation in two clinical phase II trials, seems to be a new therapeutic option for the treatment of myocardial dysfunction in sepsis associated with endotoxemia. PMID- 16369186 TI - Immune effects of resuscitation with HBOC-201, a hemoglobin-based oxygen carrier, in swine with moderately severe hemorrhagic shock from controlled hemorrhage. AB - HBOC-201, a hemoglobin-based oxygen carrier, improved physiologic parameters and survival in hemorrhagic shock (HS) animal models. However, resuscitation from HS and the properties of different fluids influence immune responses. The aim of this study was to determine if HBOC-201 significantly alters immune function in traumatic HS. Anesthetized pigs underwent soft tissue injury, controlled hemorrhage of 40% of blood volume, and resuscitation with HBOC-201 or Hextend, or no resuscitation. Sequential whole-blood samples were collected for analyses of leukocyte differential (hematology analyzer), T-lymphocyte subsets (CD3, CD4, and CD8) (FACS), lymphocyte adhesion marker CD49d (alpha4-integrin) expression (FACS), plasma cytokines-tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-10-(ELISA), and lymphocyte apoptosis (annexin-V/propidium iodide staining) (FACS). Statistical analyses were performed by the mixed procedure. Total WBC counts decreased posthemorrhage in both resuscitation groups. Lymphocyte percentages decreased and PMN percentages increased around 4 h posthemorrhage in all groups. CD3 cells decreased in all groups, but CD4 and CD8 cells decreased only in the resuscitation groups. TNF-alpha levels were not detectable in any groups. IL-6 levels were similar across treatment groups (P > 0.05); however, IL 10 levels were higher in the HBOC group, as early as 1 h posthemorrhage (P = 0.04). Increases in lymphocytic CD49d expression levels and apoptosis occurred only in nonresuscitation and Hextend groups, respectively (P < or = 0.01). In comparison with Hextend, HBOC-201 had no significant adverse or beneficial effects on immune function in this model of moderately severe HS in swine, suggesting that it may be safe as a resuscitation fluid in HS patients. PMID- 16369187 TI - Parenteral N-3 fatty acids modulate inflammatory and immune response in rats undergoing total gastrectomy. AB - This study investigated the effect of n-3 fatty acid (FA)-containing parenteral nutrition on the circulatory lymphocyte subpopulation, intracellular cytokine and leukocyte adhesion molecule expression, and phagocytic activity in rats undergoing total gastrectomy. Normal rats with internal jugular catheters were assigned to normal control (NC) and two experimental groups and received total parenteral nutrition (TPN). At the same time, a total gastrectomy was performed in the experimental groups, whereas the NC group underwent a sham operation. The TPN solutions were isonitrogenous and identical in nutrient compositions except for differences in fat emulsion contents. The NC and one of the experimental groups received a soybean oil emulsion (SO), and the other experimental group received 50% soybean oil and 50% fish oil emulsion (FO). Half of the rats in each respective group were sacrificed 1 or 3 days after surgery or the sham operation to examine their immune response. The results showed that the FO group had a higher CD4 proportion and CD4/CD8 ratio than those of the SO and NC groups postoperatively. The phagocytic activity of peritoneal macrophages was higher in the FO group than in the NC group, but no difference was found between the SO and NC groups 3 days after surgery. The intracellular interferon (IFN)-gamma distribution in the FO group was higher than that of the SO group on postoperative days. Leukocyte adhesion molecule expressions and peritoneal monocyte chemotactic protein-1 levels were lower in the FO group than in the SO group on postoperative day 3. These results suggest that parenterally infused FO did not result in immunosuppression. In addition, FO administration promotes lymphocyte Th1 cytokine production, enhances peritoneal macrophage phagocytic activity, and reduces leukocyte adhesion molecule expression in rats with total gastrectomy. PMID- 16369188 TI - B lymphocytes undergo apoptosis because of FcgammaRIIb stress response to infection: a novel mechanism of cell death in sepsis. AB - Sepsis is predominantly characterized by proinflammatory signs in its initial phase, but can be also associated with immune suppression that can be a consequence of apoptotic cell death. The role of Fc receptors (FcRs) is poorly understood in this disease, and it was recently shown that, in addition to the promotion of opposite inflammatory responses, they are implicated in apoptosis. Using a model of peritonitis in mice that do not express activating FcRs, we tested the hypothesis that FcgammaRIIb, the only known immunoglobulin G receptor capable of inducing apoptosis, would participate in the induction of this kind of cell death during serious infection. The blocking of this receptor by a monoclonal antibody significantly decreased the number of apoptotic splenic B cells, demonstrating its involvement in apoptosis. FcgammaRIIb-mediated apoptosis was neither the result of increased TNFalpha levels nor was it associated with IL 10 production. Finally, the decreased apoptosis after mice treatment with FcgammaRIIb-blocking antibody was not sufficient to increase its survival. Thus, we conclude that although apoptosis is a multifactorial phenomenon in sepsis, one of these factors is the inhibitory immunoglobulin G receptor FcgammaRIIb. FcgammaRIIb stress response to infection is a novel mechanism that contributes to the comprehension of apoptosis in sepsis. PMID- 16369189 TI - Prevention of ischemia reperfusion injury by positive pulmonary venous pressure in isolated rat lung. AB - Pulmonary ischemia-reperfusion (I/R) without tissue hypoxia induces inflammatory cytokine mRNA expression in the lung under the condition of 0 mm Hg pulmonary venous pressure (0PVP), which might be a cause of I/R injury. Our aim is to determine whether the pulmonary vascular endothelium expresses cytokine mRNAs and their corresponding proteins or develops I/R injury when positive PVP is maintained during ischemia to provide a positive stretch to the endothelium throughout the ischemic period. In isolated, perfused, and ventilated rat lungs, the right and left pulmonary arteries were isolated, and the left lung was selectively occluded for 60 min and then reperfused for 30 min. During ischemia, the left atrial pressure was maintained at 5 mm Hg (5PVP) or 0PVP. TNF-alpha, IL 1beta, IL-6, and IL-10 mRNA expression in the lungs was evaluated by RT-PCR and in situ hybridization, and the production and localization of corresponding proteins were determined by staining with fluorescence-labeled antibodies against the cytokines and an antibody against CD34. Pulmonary vascular/epithelial permeability was evaluated by measuring albumin content in bronchoalveolar lavage (BAL) fluid and wet/dry ratio. At 5PVP, there were no increases in the left lung perfusion pressure, albumin content in BAL fluid, wet/dry ratio, or expression of cytokine mRNAs and their corresponding proteins on the vascular endothelium by I/R. In contrast, at 0PVP, the increased expression of cytokine mRNAs and their corresponding proteins on the vascular endothelium by I/R was verified. The finding that the application of 5PVP during ischemia abolished the expression of cytokine mRNAs and their corresponding proteins as well as the I/R injury gives us new insights in the study of lung preservation for transplantation. PMID- 16369190 TI - Intravenous administration of choline or cdp-choline improves platelet count and platelet closure times in endotoxin-treated dogs. AB - This study was performed to assess the effects of intravenous choline chloride and cytidine-5'-diphosphate choline (CDP-choline) treatments on circulating platelet, white blood cell, and red blood cell counts and platelet functions in response to endotoxin. Saline (0.2 mL/kg), choline chloride (20 mg/kg), or CDP choline (70 mg/kg) were given intravenously three times at 4-h intervals, and endotoxemia was induced by endotoxin (E. coli 055:B5, 20 microg/kg) infusion, 5 min after the first treatment. Blood samples were collected before and at multiple time points after the challenge, for a panel of hematologic parameters and platelet closure times measured by PFA-100. In saline-treated dogs, circulating platelet counts decreased by 85% (P < 0.001) at 0.5 h and remained low by 36%-80% (P < 0.5-0.001) 1-12 h after endotoxin. Circulating WBC counts decreased by 80%-90% (P < 0.001) at 0.5-2 h, and increased (P < 0.001) by 190% 12 h after the endotoxin. In response to endotoxin, RBCs increased by 10%-13% (P < 0.05) at 1-12 h. Endotoxin-induced decline in circulating platelets was attenuated at 0.5 h (P < 0.05-0.01) and reversed at 1-12 h (P < 0.05-0.001) by choline. Platelet closure times were shortened from 81 +/- 10 s and 135 +/- 10 s to 29 +/- 5 s (P < 0.001) and 60 +/- 3 s (P < 0.001) at 0.5 h, and prolonged (P < 0.001) at 1-8 h after endotoxin induction. Endotoxin-induced shortening in platelet closure times was attenuated (P < 0.05) and blocked (P < 0.01) by choline and CDP-choline, respectively. These results showed that choline and CDP choline treatments improved circulating platelet counts and platelet function during endotoxemia in dogs. PMID- 16369191 TI - Fibronectin prevents D-galactosamine/lipopolysaccharide-induced lethal hepatic failure in mice. AB - Plasma fibronectin (FN) has a broad range of biological functions involved in cellular adhesion, motility, differentiation, apoptosis, hemostasis, wound healing, reticuloendothelial system function, and ischemic injury. In this study, we examined the effects of FN on D-galactosamine (GalN)/lipopolysaccharide (LPS) induced fulminant liver failure in mice. Female Balb/c mice received intraperitoneal injection of 50 mug/kg of LPS and 400 mg/kg of GalN simultaneously. Thirty minutes before GalN/LPS administration, human plasma FN (FN group) or the same dose of human serum albumin (control group) was given intravenously. GalN/LPS induced a marked decrease in plasma FN, which was reversed by FN pretreatment. The survival rate of the FN group was markedly improved in a dose-dependent manner compared with that of the control group (survival rate 0%). FN prevented increases in the concentrations of serum enzymes and total bilirubin related to liver injury. FN pretreatment significantly suppressed tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and interleukin (IL)-6 levels, and enhanced IL-10 levels in serum and liver tissue compared with the control group. Moreover, TUNEL staining, caspase 3 and 8 activities, and necrosis in the remnant liver were significantly decreased in the FN-treated rats compared with the controls. Furthermore, FN pretreatment inhibited the activation of nuclear factor (NF)-kappaB and increased the expression of Bcl-xL protein in liver tissue. These results suggest that FN protected against GalN/LPS-induced liver failure by a mechanism involving inhibition of NF-kappaB activation, which caused down-regulation of TNF-alpha and involved up-regulation of IL-10, and elevation of Bcl-xL induced a blockage of apoptotic signals, by which apoptosis of hepatocytes caused by GalN/LPS was suppressed. PMID- 16369192 TI - Mannose-binding lectin polymorphisms in severe sepsis: relationship to levels, incidence, and outcome. AB - Mannose-binding lectin (MBL) genetic polymorphisms result in deficiency of the encoded protein and increased susceptibility to infection, especially in children and the immunocompromised. The objective of this study was to investigate the relationship between MBL-2 exon 1 and promoter -221 polymorphisms, plasma levels of the encoded protein, and the incidence and outcome of severe sepsis and septic shock. One hundred seventy-four white adult patients with severe sepsis or septic shock were recruited in a prospective multicenter study across eight intensive care units in the South of England, UK. Genotype and haplotype frequencies were compared between normal population controls and patients, and between survivors and nonsurvivors. Plasma levels of encoded protein were related to genotype and outcome. The exon 1 polymorphisms (A/O or O/O) were significantly more common in the patients with severe sepsis and septic shock than in normal healthy adults (54.6% vs. 39.7%, P = 0.001), and there was a significant difference in haplotype frequency between controls and septic patients (P < 0.0001). There was no significant difference in MBL-2 genotype or haplotype frequency between survivors and nonsurvivors. There was a strong relationship between MBL-2 haplotype and plasma MBL concentration (P < 0.001). Individual plasma levels were variable and increased between days 1 and 7. The mortality rate was higher in those with MBL levels <1000 microg/L than in those patients with levels >1000 microg/L (47.2 vs. 22.2%, P = 0.05). We conclude that genetic polymorphisms resulting in mannose binding lectin deficiency are associated with increased susceptibility to sepsis. The close relationship between polymorphic variants and plasma MBL concentration persists during sepsis but individual levels vary widely. Lower circulating MBL levels are associated with a poor outcome. PMID- 16369193 TI - Vitamin D endocrine system and the genetic susceptibility to diabetes, obesity and vascular disease. A review of evidence. AB - The Vitamin D endocrine system regulates multiple aspects of calcium metabolism and cellular differentiation and replication in the immune system, endocrine pancreas, liver, skeletal muscles and adipocytes. It plays an important role in glucose homeostasis, notably, in the mechanism of insulin release. Actions of vitamin D are mediated by the binding of 1, 25-(OH)2D3 to a specific cytosolic/nuclear vitamin D receptor (VDR), a member of the steroid/thyroid hormone receptor superfamily. Several frequent polymorphisms are found in the VDR gene and were reported to be associated with a variety of physiological and pathological phenotypes in many populations. In this paper, we will review the evidences suggesting associations of allelic variations in the VDR gene and phenotypes related to body weight, glucose homeostasis, diabetes and its vascular complications. PMID- 16369194 TI - Impact of high- and low-intensity targeted exercise training on the type of substrate utilization in obese boys submitted to a hypocaloric diet. AB - BACKGROUND: We assessed the effect of two programs combining a hypocaloric diet with low-intensity (LI) or high-intensity (HI) exercise training, during two months, on substrate utilization at exercise in obese children. METHODS: Fifteen obese boys participated in a combined program of exercise and caloric restriction induced weight loss (diet starting two weeks before the training program). The maximal fat oxidation point (Lipox max) was determined to individualize exercise training. Training consisted of cycling at either LI (Lipox max) for seven children or HI (Lipoxmax+40% Lipox max) for eight children. RESULTS: All children exhibited a decrease in weight (LI: -5.2 kg +/- 0.7 (P<0.01), HI: -7 kg +/- 0.7 (P<0.01)). While in the LI group, both fat and CHO oxidation were unchanged after training, HI group oxidize less fat and more CHO after training when exercising at 20% and 30% Wmax th (P = 0.02). DISCUSSION: While a LI exercise training program maintains (but does not improve) the ability to oxidize fat at exercise, HI training actually shifts towards CHO the balance of substrate oxidation during exercise. Thus, a low intensity training protocol seems to counteract to some extent the decline in lipid oxidation at exercise that occurs after a hypocaloric diet, and is thus likely to be synergistic to diet in the weight lowering strategy. PMID- 16369195 TI - Taurine modifies insulin signaling enzymes in the fructose-fed insulin resistant rats. AB - OBJECTIVES: High fructose feeding induces insulin resistance and hyperinsulinemia in rats. The present study was proposed to elucidate the derangements in the insulin signaling pathway in high fructose-fed rats and whether taurine, a sulphur-containing amino acid could improve insulin action by modulating the signal transduction pathway. METHODS: Male Wistar rats of body weight 170-190 g were divided into 4 groups of 6 rats each. Control rats received control diet and water ad libitum. Fructose fed animals received high fructose diet (> 60% of total calories) and water ad libitum. Fructose + taurine rats received fructose diet and 2% taurine solution ad libitum. Control + taurine rats received control diet and 2% taurine solution ad libitum. After the experimental period of 30 days, the effects of taurine on certain parameters on glucose metabolism were determined. The activities of protein tyrosine kinase (PTK) and protein tyrosine phosphatase (PTP) were assayed in liver. RESULTS: The activities of the glycolytic enzymes were significantly lower while the activities of the gluconeogenic enzymes were higher in untreated fructose-fed rats as compared to control animals. Depletion of liver glycogen was observed in fructose-fed rats. Fructose-fed rats showed alterations in the activities of insulin signaling enzymes PTK and PTP. Taurine administration improved insulin sensitivity and controlled hyperglycemia and hyperinsulinemia in fructose-fed rats. Taurine treatment also restored the glucose metabolizing enzyme activities in fructose fed rats. CONCLUSIONS: Taurine supplementation might have a beneficial effect in overcoming insulin resistance and its associated abnormalities by modifying the post-receptor events of insulin action. PMID- 16369196 TI - Nocturnal differences in subcutaneous tissue glucose between forearm and abdominal sites during continuous glucose monitoring in normal subjects. AB - OBJECTIVE: A number of short-term studies using the continuous glucose monitoring system (CGMS) indicate that improved metabolic control can be observed in patients with type 1 diabetes when CGMS is applied in clinical practice. Data have also accumulated to suggest that spot measurements of glucose performed four times a day would not detect as much as 70% of all hypoglycaemic episodes registered by CGMS. When more frequent reference values were obtained however it was inferred that nighttime hypoglycaemia reported by CGMS may be spurious. As most assessments with CGMS have been utilizing abdominal subcutaneous tissue, we were interested to evaluate whether differences between blood glucose and sensor readings obtained from different sites exist. RESEARCH DESIGN AND METHODS: Two viscometric affinity glucose sensors, applicable to subcutaneous tissue of both forearm and abdomen, were inserted subcutaneously in 12 non-diabetic subjects. Sensors generated glucose data at 3 min intervals and venous blood glucose was determined in duplicates by HemoCue at 15-90 min intervals for 24 hours. Each subject consumed three carbohydrate-rich meals, performed an exercise test, and was observed during nocturnal bed-rest at the research center. RESULTS: The initial decrease of blood glucose during exercise was not fully detected by the sensors. Otherwise, no significant differences between sensor values and blood glucose were observed during day-time. During nocturnal bed-rest abdominal sensor values came approximately 20% lower than blood glucose (P<0.001) and forearm sensor readings. CONCLUSION: It is concluded that a difference between glucose values obtained from abdominal and forearm subcutaneous fat can be observed during nocturnal bed-rest in non-diabetics. PMID- 16369197 TI - Epidemiology of the metabolic syndrome in 2045 French military personnel (EPIMIL study). AB - CONTEXT: The Metabolic syndrome is considered to be an important public health problem, but few epidemiological studies have defined the present situation in France. EPIMIL is a prospective epidemiological study that began on February 1, 2003. SUBJECTS AND METHODS: This study was designed to identify the clinical and laboratory parameters of metabolic syndrome and cardiovascular risk factors in a population of 2045 male military personnel based in the Paris region. The initial 1-year cross-sectional study will be followed by a 10-year follow-up and patient care. RESULTS: The 2045 subjects included 185 (9%) presented at least 3 of the 5 NCEP ATP III criteria defining metabolic syndrome. They were significantly older (42.2 +/- 8.5 yrs) than the other subjects (37.3 +/- 8.7 yrs, P<0.001), had a higher BMI (29.5 +/- 3.4 vs 24.8 +/- 2.9 kg/m2, P<0.001) and a greater body weight at age 20 (75.4 +/- 11 vs 70.4 +/- 8.5 kg, P<0.01). Smoking, little physical activity and family histories of diabetes and arterial hypertension were more frequent in these men. While levels of the cholesterol and CRP us were higher, Lp(a) and homocysteine concentrations were normal. Plasma insulin and BMI (r = 0.456 P<0.0001) and plasma insulin and waist circumference (r = 0.446 P<0.0001) were well correlated. Lastly, plasma insulin, free fatty acids and cardiovascular risk increased steadily with the increase in the number of metabolic syndrome criteria. CONCLUSIONS: These results in a large and particularly uniform population of men show the prevalence of metabolic syndrome in adult men, and demonstrate its link with insulin resistance. Men with several risk factors require specific care, particularly for hypertension and dyslipidemia; the effectiveness of this care will be evaluated during the follow up period. PMID- 16369198 TI - Gestational diabetes mellitus and the risk of metabolic syndrome: a population based study in Lausanne, Switzerland. AB - AIMS: To investigate the relationships between gestational diabetes mellitus (GDM) and the metabolic syndrome (MS), as it was suggested that insulin resistance was the hallmark of both conditions. To analyse post-partum screening in order to identify risk factors for the subsequent development of type 2 diabetes mellitus (DM). METHODS: A retrospective analysis of all singleton pregnancies diagnosed with GDM at the Lausanne University Hospital for 3 consecutive years. Pre-pregnancy obesity, hypertension and dyslipidaemia were recorded as constituents of the MS. RESULTS: For 5788 deliveries, 159 women (2.7%) with GDM were identified. Constituents of the MS were present before GDM pregnancy in 26% (n = 37/144): 84% (n = 31/37) were obese, 38% (n = 14/37) had hypertension and 22% (n = 8/37) had dyslipidaemia. Gestational hypertension was associated with obesity (OR = 3.2, P = 0.02) and dyslipidaemia (OR = 5.4, P=0.002). Seventy-four women (47%) returned for post-partum OGTT, which was abnormal in 20 women (27%): 11% (n = 8) had type 2 diabetes and 16% (n = 12) had impaired glucose tolerance. Independent predictors of abnormal glucose tolerance in the post-partum were: having > 2 abnormal values on the diagnostic OGTT during pregnancy and presenting MS constituents (OR = 5.2, CI 1.8-23.2 and OR = 5.3, CI 1.3-22.2). CONCLUSIONS: In one fourth of GDM pregnancies, metabolic abnormalities precede the appearance of glucose intolerance. These women have a high risk of developing the MS and type 2 diabetes in later years. Where GDM screening is not universal, practitioners should be aware of those metabolic risks in every pregnant woman presenting with obesity, hypertension or dyslipidaemia, in order to achieve better diagnosis and especially better post-partum follow-up and treatment. PMID- 16369199 TI - Percutaneous transluminal angioplasty in severe diabetic foot ischemia: outcomes and prognostic factors. AB - OBJECTIVES: To evaluate the outcomes of severe ischemic diabetic foot ulcers for which percutaneous transluminal angioplasty (PTA) was considered as the first line vascular procedure. Factors associated with successful PTA were sought. RESEARCH DESIGN AND METHODS: In 32 consecutive diabetic patients with foot ulcers and severe limb ischemia, PTA was performed if feasible; if not, primary bypass grafting was done when feasible. All patients were followed until healing or for at least one year. Patients with worsening ulcers after PTA underwent bypass grafting. Clinical and angiographic factors influencing outcomes after PTA were sought by univariate and multivariate analysis. RESULTS: PTA was done in 25 of the 32 (78%) patients, and considered clinically successful in 13 (52%). After 1 year, the healing rate was 70% and the limb salvage rate 90%. Successful PTA was significantly associated with a higher post-PTA transcutaneous oxygen pressure (P = 0.03) and presence of at least one patent pedal vessel (P = 0.03) in the univariate analysis; only a patent pedal vessel was significant in the multivariate analysis. CONCLUSION: Primary PTA in diabetic patients with severe ischemic foot ulcers provides similar outcomes to usual results obtained in severe ischemia in absence of diabetes. The presence of one patent pedal vessel on arteriography before PTA is the best prognostic factor. PMID- 16369200 TI - Effectiveness of patient education on glycemic control in insulin treated patients in general practice. AB - OBJECTIVE: To assess the effectiveness of a patient education program on diabetes control. METHODS: A pre-test post-test pre experimental study was performed with 87 insulin treated diabetic patients followed in the chronic disease unit of Kelibia district (Nabeul, Tunisia). Subjects received education in 6 monthly sessions. Outcome measures included glycated haemoglobin (HbA1c), anxiety level according to the Hamilton scale and Body Mass Index. Changes were assessed after the 6 month program. RESULTS: HbA1c decreased from 8.8% +/- 1.23 to 7,6 +/- 1.43 (P<10-6) with an increase in the prevalence of satisfactory HbA1c (< or = 8%) from 33% to 61.2% (P<0,001). The prevalence of severe anxiety decreased by about a third (P < 0,001). CONCLUSION: Education led to an improvement in diabetes control in insulin treated diabetic patients. More attention should be paid to such strategies in general practice. PMID- 16369201 TI - Early decrease of the percent of HOMA beta-cell function is independently related to family history of diabetes in healthy young nonobese individuals. AB - OBJECTIVE: To determine the relationship between family history of diabetes (FHD) and decrease in percent of HOMA beta-cell function (HOMA-beta%) index in healthy betanon-obese Mexican subjects. MATERIALS AND METHODS: Forty-eight individuals (30 women and 18 men) with FHD were compared vs 48 control subjects (30 women and 18 men) in a cross-sectional study matched by age, sex, and Waist-to-Hip ratio. Pregnancy, obesity, being overweight, alcohol consumption, high blood pressure, and heavy physical activity were exclusion criteria. All the participants were required to have a Body Mass Index < 25 kg/m2 and serum fasting and 2-hours postload glucose levels lower than 6.1 mmol/l and 7.8 mmol/l, respectively. The reciprocal of serum fasting insulin concentrations (1/Ins0) (microU/ml) and HOMA B% index were used as indicators of insulin sensitivity and beta-cell function. RESULTS: Average age was of 19.4 +/- 3.6 vs 19.8 +/- 2.6, P = 0.66 for the subjects with and without FHD. HOMA-beta% index was significantly lower in the subjects with FHD (186.1 +/- 74.1 vs 252.7 +/- 149.5, P = 0.01). For similar levels of insulin sensitivity, subjects with FHD showed lower HOMA-beta% index than control subjects (P < 0.001). Multivariate regression analysis showed a strong and independent relationship between FHD and decrease of HOMA-beta% index (OR 2.6, CI95% 1.2-4.3, P = 0.01). CONCLUSIONS: This study shows that normal weight offspring of type 2 diabetes subjects exhibited a significant decrease of HOMA-beta% index suggesting that FHD exerts an independent early negative effect on beta-cell function. PMID- 16369202 TI - Active stents in diabetic patients. AB - In the treatment of coronary stenosis, evolution after PTCA is not as good in diabetic patients compared to non diabetic ones, whatever the treatment used. We now have data of large clinical studies which show good results of drug loaded stents in diabetic patients, especially with either a cytostatic drug (sirolimus) or a cytotoxic one (paclitaxol). In the RAVEL study, among the 44 diabetic patients, 19 had sirolimus stenting with a restenosis rate of 0% vs a restenosis rate of 40% for the 25 patients with standard stents. In the 279 diabetic patient group of the SIRIUS study, the restenosis rate (50% or more stenosis rate) was 17.6% when sirolimus stenting was used vs 50.5% for the patients with standard stenting and at 9 months and target lesion revascularisation was from 22.3% with bare metal stents, compared to 6.9% with sirolimus eluting stents. In the TAXUS IV study, the advantage was evident in diabetic patients with a restenosis rate 80% lower in patients treated with oral anti diabetic therapy and 82% in patients treated with insulin. In the TAXUS VI study, the target lesion revascularisation rate of diabetic patients was 2.6% when taxus MR (modified release) was used, vs 22.6% with standard stents. The event which until now made PTCA different from surgery was restenosis, especially in diabetic patients. The analysis of use of recent active stenting registries has shown that diabetic patients have now much better long term results than previously reported. PMID- 16369203 TI - Systemic allergy to human insulin and its rapid and long acting analogs: successful treatment by continuous subcutaneous insulin lispro infusion. AB - Since the introduction of highly purified human recombinant insulin, allergy to insulin has become a very rare clinical situation, encountered in less than 1% of patients. It results in potentially life-threatening immediate or delayed, local and general manifestations. Different treatments of unequal efficiency have been proposed, the use of insulin analogs showing benefits in certain situations. We report the case of a type 2 diabetic patient who presented local reactions and then an anaphylactic shock after the introduction of insulin analog premixes. Intra-dermal reactions performed with porcine, human and insulin analogs preparations (aspart, lispro, glargine) were all positive, as well as the specific anti-insulin IgE measurement. Because we could not achieve normoglycaemia with maximal oral treatment and low caloric diet, we decided to attempt a desensitisation by continuous subcutaneous infusion of insulin lispro, since the lowest skin reaction was obtained with this insulin. We were able to induce a tolerance, by means of very low basal rate, very slowly increased, without any boluses, and maintaining antihistamine therapy. Six months later, the patient remains free of any symptom and has achieved a quite good glycaemic control. We describe for the first time a case of allergy to human insulin and to all available rapid and long acting analogs. We show the interest of a treatment with CSII of analogs in order to induce tolerance. PMID- 16369204 TI - Advantages to using capillary blood beta-hydroxybutyrate determination for the detection and treatment of diabetic ketosis. AB - Ketone body determination is indicated in all diabetic patients when the risk of ketotic decompensation exists. New methods of screening for ketosis, in particular capillary blood ketone body determination, provide analytical, technical and clinical advantages compared to the conventional ketonuria. It is proposed that a diabetic patient with hyperglycaemia (capillary blood glucose > 2.50 g.l(-1)) and capillary blood ketone bodies exceeding 0.5 mmol.l(-1) requires therapeutic management. For values greater than 3 mmol.l(-1) or in case of more serious clinical symptoms, hospitalisation is indicated, considering the high probability of ketoacidotic decompensation. The advantages of capillary blood ketone body determination including easy use, and rapid and objective results may improve management of the diabetic patient, especially in emergency situations. However, prescription by a physician of capillary blood ketone body determination should be offered to targeted populations that have a high risk of ketoacidotic decompensation, after providing education to patients that is above all aimed at preventing this metabolic complication. In this context of determining ketone bodies in capillary blood, the term "capillary blood ketone bodies" is therefore preferable to the term "capillary blood beta-hydroxybutyrate determination". Indeed, it appears more appropriate, simple, descriptive and significant both for health-care staff and for patients. PMID- 16369206 TI - In memoriam: Cesar Bergada MD (1929-2005). PMID- 16369207 TI - In memoriam: John D. Crawford, II, MD (1920-2005). PMID- 16369208 TI - Growth hormone / insulin-like growth factor-1 axis during puberty. AB - Puberty is a dynamic, transitional period of life which is characterized by the acquisition of secondary sexual characteristics leading to the development of fertility. Puberty is accompanied by sexually dimorphic changes in linear growth, body proportions and body composition. The pubertal growth spurt is influenced by a number of factors such as hormones, nutrition, physical activity and general health, acting mostly in concert in order to modify a genetic potential for growth. Growth hormone (GH) is traditionally considered to be the main regulator of growth. During puberty, elevated sex steroid concentrations (especially oestrogens) stimulate GH production, leading to an activation of the whole GH/Insulinlike growth factor-1 (IGF-1) axis. This activation is mostly characterized by an increase in the amplitude of GH pulses rather than an increase in frequency or in duration. Interactions between GH and sex steroids (especially androgens) express an anabolic effect on muscle mass, bone mineralization and body proportion which constitutes the male and the female adult body composition. PMID- 16369209 TI - Metabolic effects of IGF-I deficiency: lessons from mouse models. AB - Insulin and insulin-like growth factors (IGFs) belong to the most biologically characterized family of peptides involved in metabolism, growth and development. The cellular responses to the IGFs are mediated primarily by the IGF-I receptor. The IGF-I receptor is a member of the family of tyrosine kinase growth factor receptors, and is highly homologous (70%) to the insulin receptor, especially in the tyrosine kinase domain (84%) ADDIN. Upon ligand binding to the extracellular region, the intrinsic tyrosine kinase domain of the receptor is activated. In the past it was believed that insulin activates primarily metabolic processes while IGFs promote cell growth and differentiation. However, in the last two decades many animal models of IGFI deficiency and excess revealed the importance of IGF-I in carbohydrate and lipid metabolism and now it is clear that these peptide hormones together with growth hormone (GH) work in a coordinate and interdependent manner. In the circulation, IGFs are bound in a binary complex with a family of high affinity IGF-binding proteins (IGFBPs) ADDIN. However, most of the circulating IGF-I associates with a high molecular weight complex approximately 150 KDa consisting of IGFBP-3 and the acid labile subunit (ALS) ADDIN. Once the ternary complex dissociates, the binary complexes of IGFBP-IGFs are removed from the circulation and by crossing the endothelium to reach the target tissues and to interact with cell surface receptors. In the present review we will summarize the role of GH and IGF in somatic growth and focus on the metabolic effects of IGF-I deficiency as assessed in various mouse models. PMID- 16369210 TI - Genetic susceptibility in thyroid autoimmunity. AB - The autoimmune thyroid diseases (AITD) include Graves' disease (GD) which manifests in hyperthyroidism and Hashimoto's thyroiditis (HT), manifesting as hypothyroidism. Genetic susceptibility in combination with external factors (e.g. dietary iodine) are believed to initiate the autoimmune response to thyroid antigens in AITD. Indeed, there is solid epidemiological data to support a strong genetic influence on the etiology of AITD including family and twin studies. Recently, there has been significant progress toward the identification of the AITD susceptibility genes. Several loci (genetic regions) that are linked with AITD have been mapped and in some of these loci putative AITD susceptibility genes have been identified. Some of these loci predispose to a single phenotype (GD or HT), while other loci are common to both diseases, indicating that there is a shared genetic susceptibility to GD and HT. The putative GD and HT susceptibility genes include both immune modifying genes (e.g. HLA, CTLA-4) and thyroid specific genes (e.g. TSHR, Tg) and it is likely that the final disease phenotype is a result of an interaction between these loci, as well as environmental influences. PMID- 16369211 TI - How should we treat type 2 diabetes in youth? AB - The worldwide increase of type 2 diabetes in youth is a critical problem. It is important to prevent the development of type 2 diabetes in high-risk individuals. In many patients with type 2 diabetes, hyperglycemia can be reduced with appropriate changes in diet and exercise. However, some patients with persistent HbA1c levels >7.5% need pharmacological therapy to improve their metabolic control. A variety of oral hypoglycemic agents, including alpha-glucosidase inhibitors, sulfonylureas and metformin, are available. Metformin is widely used in pediatric patients and is considered to be the most effective oral agent. In some cases, combination therapy with metformin and sulfonylureas or use of insulin is more effective to stabilize glycemia. The approach to insulin therapy in type 2 diabetes often differs from that used in type 1 diabetes. The therapeutic approach to childhood type 2 diabetes should be individually tailored. PMID- 16369212 TI - The art and science of regression modelling; methods for building valid models to explore hormone and body composition interactions. AB - Multiple linear regression modelling is commonly used to investigate how hormones and body composition interact, but for valid interpretation a sound methodological approach must be used. It is particularly important that the assumptions for regression are met so that spurious associations are not generated. In this article we show how different approaches to building a multiple linear regression model can influence perceived associations, using examples from the literature and our own data related to predicting fasting insulin and leptin levels from total body fat and fat distribution in children. PMID- 16369213 TI - HIV--associated lipodystrophy in children. AB - Lipodystrophy (LD) with varying degrees of lipohypertrophy, lipoatrophy, hyperlipidemia, and insulin resistance is one of the complications of highly active antiretroviral therapy (HAART) and occurs in one to 33 % of HAART-treated, HIV infected children. We summarize the data on the role of leptin, adiponectin, the growth hormone axis, glucocorticoids, sterol response element binding protein 1c (SREBP-1c), the tumor necrosis factor alpha axis (TNF-alpha), interleukin-6 (IL-6), interleukin- 18 (IL-18), interferon-alpha (IFN-alpha), tissue plasminogen activator (tPA), and plasminogen activator inhibitor (PAI-1) in the pathophysiology of LD. Adiponectin levels are generally decreased in LD, whereas leptin levels are increased. Systemic cortisol levels are not elevated in LD, even though glucocorticoids seem to play an important role in LD and the phenotype can be reminiscent of Cushing syndrome. GH resistance in LD needs to be better characterized. While some cytokines show promise as markers for LD, it is difficult to tell whether their derangement is a cause of or the effect of LD. PMID- 16369214 TI - Antibiotic prophylaxis in severe acute pancreatitis: the never-ending controversy. AB - Infections are the most common late cause of morbidity and mortality in severe acute pancreatitis. Between 1993 and 2004, a number of prospective controlled trials, meta-analyses, and guidelines all concluded that antibiotics were indicated for the prophylaxis of infections in severe acute pancreatitis. However, in 2004, the first prospective, double-blind, placebo-controlled trial was published showing no effect of prophylactic antibiotics, thus rekindling the controversy surrounding this subject. For those physicians who choose to use prophylactic antibiotics, their use should be limited to patients with pancreatic necrosis, and the choice of antibiotic should be guided by the drug's proven ability to penetrate into the pancreas. Evidence supports imipenem, meropenem, or cefepime for this indication. One third of patients will develop infection of pancreatic necrosis despite the use of prophylactic antibiotics. Vigilance must be maintained for evidence of recurrence of markers of inflammation or persistence beyond the first week, which would suggest the need for fine-needle aspiration and culture to detect infection. Physicians who choose not to use prophylactic antibiotics for pancreatic necrosis should be aware that they will need to initiate antibiotics in 54% to 77% of patients for either infection of pancreatic necrosis or extrapancreatic infections. PMID- 16369215 TI - Role of liver biopsy in the management of chronic liver disease: selective rather than routine. AB - Liver biopsy has historically played a central role in the diagnosis and management of a variety of chronic liver diseases. However, as the understanding of disease pathology has progressed, and laboratory diagnostics, imaging technology, and clinical algorithms to determine both the etiology and presence of fibrosis have advanced, the role of liver biopsy has become more circumscribed. In chronic liver disease, liver biopsy is now more often used selectively, rather than routinely, for diagnostic purposes. Newer treatment of chronic hepatitis B and C has become more effective and thus reduced the routine need to acquire tissue. Risk factors for nonalcoholic fatty liver disease are readily identified and suggest the diagnosis after exclusion of alternative considerations, and there is no specific treatment for this condition; thus there is little role for the routine use of liver biopsy to guide treatment. Only in select cases of chronic hepatitis C, especially in patients with genotype 1, an indeterminate stage and grade of disease on noninvasive evaluation, or in those with human immunodeficiency virus coinfection, for whom the risks and benefits of treatment are less clear, is there a role for routine pretreatment biopsy. PMID- 16369216 TI - Endoscopic therapy of benign biliary strictures. AB - Benign biliary strictures result from many etiologies, including postoperative injuries, anastomotic strictures, and chronic pancreatitis. Therapy is generally required to relieve symptoms of cholestasis or cholangitis and prevent secondary cirrhosis. Surgery has been the traditional method for treatment of benign biliary strictures but is associated with significant morbidity and variable long term outcomes. More research is needed to define the best methods and materials for various patient populations. Endoscopic management, including stricture dilation and stent placement, is more appealing because it is less invasive and better tolerated, but repeated treatments are required. Outcomes of endoscopic management depend on both the etiology and the location of the stricture. Accumulating data suggest that long-term success is substantially greater when multiple rather than single plastic stents are placed. Uncovered metallic stents are problematic and generally not recommended. Emerging alternatives to plastic stents include the placement of removable covered metallic stents. PMID- 16369217 TI - Ulcerative colitis therapy: importance of delivery mechanisms. AB - Since the initial observation that mesalamine or 5-aminosalicylate (5-ASA) has an anti-inflammatory effect on ulcerative colitis, investigators have been trying to improve on the delivery mechanisms of this compound. As it is believed that the anti-inflammatory effect of 5-ASAs is mediated topically, current formulations are designed to release 5-ASA in the small intestine and colon, or predominantly in the colon. A dose-response curve is seen with some preparations of mesalamine but not all. In general, 5-ASAs are effective in patients with ulcerative colitis and much less effective in Crohn's disease. Evidence demonstrates that 5-ASAs are effective for induction of remission and maintenance of remission. Preparations that deliver 5-ASA in a pH-dependent manner are most affected by variability in luminal pH, whereas those that depend on bacterial cleavage for release of the active 5-ASA are most affected by transit time. Most studies have not compared different preparations of mesalamine and examined differences in colonic delivery. Depending on the endpoint examined in the studies, efficacy of the various 5-ASA products appears similar at the most optimal doses. For a given patient, however, it may be necessary to experiment with more than one preparation if an initial trial results in a suboptimal response. PMID- 16369218 TI - Treatment of chronic hepatitis B with peginterferon. PMID- 16369219 TI - Update on CT colonography (virtual colonoscopy) trials. PMID- 16369220 TI - Intestinal pseudomembranes: Staphylococcus aureus or Clostridium difficile or both? PMID- 16369222 TI - Gastroesophageal reflux disease and sleep disorders: a wake-up call for physicians and their patients. AB - Sleep's implications to the physiology and pathogenesis of a number of diseases have been ignored until recently. With the evolution of studies conducted in sleep laboratories, the relationship between various gastrointestinal diseases, in particular gastroesophageal reflux disease (GERD), and sleep disorders is being recognized. This article discusses the personal and societal impact of GERD related sleep disorders, including quality of life issues and work and leisure impairment. A review of intervention studies indicates that GERD-related sleep disorders respond effectively to acid-suppressive medical therapy. Clinicians are advised to take a proactive stance in evaluating patients with GERD for unrecognized sleep disorders and ascertaining whether patients with sleep disorders have GERD symptoms as well. PMID- 16369223 TI - Increasing compliance with long-term therapy: avoiding complications and adverse events. AB - Compliance with long-term therapy is an important consideration in any chronic disease. Patients with gastroesophageal reflux disease (GERD) frequently stop and start treatment based on their symptoms. This form of therapy ("on-demand treatment") has been shown to be effective in patients with non-erosive reflux disease and may reduce the costs associated with long-term therapy. However, continuous medical therapy for erosive esophagitis is associated with fewer relapses and better outcomes. Similarly, long-term therapy administered continuously reduces complications such as stricture and may decrease the development of dysplasia in Barrett's esophagus. PMID- 16369224 TI - Intravenous proton pump inhibitor therapy: a rationale for use. AB - Proton pump inhibitors (PPIs) are used widely in the management of acid-related disorders and, for the majority of patients, oral therapy is highly effective. Not all patients with acid-related disorders respond completely to standard, once daily PPI therapy, but most nonresponders will generally respond to an increase in the dose or frequency of PPI therapy. At equivalent doses, oral and intravenous (IV) PPIs produce comparable acid suppression; thus there are very few clinical indications for IV PPI therapy. IV PPIs are an appropriate substitute for oral PPIs, at an equivalent dose, for patients with, for example, gastroesophageal reflux disease, peptic ulceration, or Zollinger-Ellison syndrome, who cannot take oral medication. For patients with nonvariceal, upper gastrointestinal hemorrhage, profound acid suppression (gastric pH . 6.0) optimizes clot stability and reduces the risk of rebleeding; this is achieved most effectively with an initial IV PPI bolus followed by a continuous infusion. High-dose, IV PPI therapy is beneficial and cost-effective in patients who have a high-risk lesion at endoscopy and it should be preceded by effective endoscopic hemostasis if possible. IV PPIs, preoperatively and in the intensive care setting, effectively reduce gastric acidity, but there are no convincing data that this confers any significant clinical benefit compared with other therapeutic strategies. PMID- 16369225 TI - Gastroesophageal reflux disease and extraesophageal disease. AB - The patient with extraesophageal manifestations of gastroesophageal reflux disease presents a clinical challenge. Symptom presentation overlaps with other otolaryngologic and pulmonary disease, and heartburn might be infrequent or absent. Endoscopy and pH monitoring are insensitive and therefore not useful in many patients as diagnostic modalities. Thus, antisecretory therapy is used as both a diagnostic trial and as therapy in the majority. Attention to optimizing therapy and judicious use of endoscopy and reflux monitoring are needed to minimize cost and maximize success. PMID- 16369226 TI - Nonsteroidal anti-inflammatory drugs, aspirin, and gastrointestinal prophylaxis: an ounce of prevention. AB - Because of the widespread use of nonsteroidal anti-inflammatory drugs (NSAIDs), NSAID-related gastrointestinal (GI) complications are recognized as the most prevalent drug toxicity in the United States. The withdrawal of 2 selective cyclooxygenase-2 (COX-2) inhibitors (agents specifically developed to reduce ulcer complications) from the US market owing to adverse cardiovascular events has made reducing ulcer risk more important than ever for patients receiving NSAID treatment. In addition, the impact of aspirin, implicated by itself as a cause of serious adverse GI events and in combination with NSAIDs as a source of incremental risk, remains under-appreciated. Balancing the cardiovascular risks of selective COX-2 inhibitors with the higher GI risks associated with conventional NSAIDs remains a major clinical challenge. Gastro-protective therapy, such as with a proton pump inhibitor, is beneficial in patients receiving NSAIDs, but despite current treatment recommendations, this "ounce of prevention" remains substantially underused for patients at risk. PMID- 16369228 TI - Periodontal disease among diabetics in Iraq. AB - BACKGROUND: It is generally accepted that diabetes increases the prevalence and severity of periodontitis, and is now considered the sixth "-opathy" of diabetes. The aim of this study was to determine the prevalence of periodontal disease in patients with type 2 diabetes mellitus and assess their severity in a single center in Basrah, Iraq. METHODS: This was a cross-sectional, case-control study. A total of 1593 patients were included (633 diabetic patients and 960 nondiabetics as controls). Patients with type 2 diabetes mellitus who were dentate persons aged 40 years and older and with = 6 remaining teeth were studied. RESULTS: Periodontal disease was seen in 95.9% of both diabetics and controls. Concerning mild and advanced disease, there was no difference between diabetics and controls, whereas moderate disease was more prevalent in those with diabetes. CONCLUSION: Of both the diabetics and controls, periodontal disease was seen in 95.9%. Moderate disease was significantly more common among diabetics. Larger studies, with pocket depth--attachment loss with radiologic evaluation for bone loss--are needed to assess the prevalence of periodontal disease among diabetics, other systemic disease, and the general population. PMID- 16369227 TI - Effect of in vitro glucose and diabetic hyperglycemia on mouse kidney protein synthesis: relevance to diabetic microangiopathy. AB - To test the possible roles of diabetic hyperglycemia, we studied the in vitro effect of increasing glucose concentrations (5.0-27.5 mmol/L) on protein synthesis (PS) of the kidneys from "adult" male albino Swiss mice. In mouse kidney cortex slices, PS (3H-leucine incorporation into trichloroacetic acid precipitable material), measured as cpm/mg protein/45 minutes, was already stimulated by 5.0 mmol/L of glucose (+24%, P < .05). At supraphysiologic glucose concentrations, PS was stimulated by 48% at 8.8 mmol/L of glucose and 31% at 13.6 mmol/L of glucose (P < .05, compared with the value observed at 5.0 mmol/L of glucose). However, the highest glucose levels (15.4 mmol/L and 27.5 mmol/L) were no longer effective. Other substrates (1.25 mmol/L or 6.26 mmol/L palmitic acid and 100 mcmol/L sorbitol) were without effect. Similar results were obtained when data were expressed as cpm/mg DNA/45 minutes. In contrast to adult mice, "young" mice showed the maximum stimulatory effect (+86%, P < .02), with a glucose concentration still in the nondiabetic range (6.6 mmol/L). However, in the "older" mice maximum stimulation was observed in the presence of high glucose concentrations (15.4 mmol/L and 27.5 mmol/L) with 52% (P < .02) and 26% (P < .05) increases, respectively, vs the value recorded at 5 mmol/L of glucose. With regard to the in vivo effect of diabetic hyperglycemia, the renal PS of 3-day streptozotocin diabetic mice was moderately increased, whereas the liver PS was markedly reduced. The effects of in vitro glucose and in vivo diabetic hyperglycemia, as modulated by both the concentration of glucose and the age, may lead to diabetic renal hypertrophy and the increased formation/accumulation of glycoproteins, thus contributing to microangiopathy. PMID- 16369230 TI - Medical investigations of homicides of prisoners of war in Iraq and Afghanistan. PMID- 16369229 TI - LDL-C goal attainment with ezetimibe plus simvastatin coadministration vs atorvastatin or simvastatin monotherapy in patients at high risk of CHD. AB - AIMS: To compare the proportion of patients at high risk for coronary heart disease (CHD) achieving the recommended low-density lipoprotein cholesterol (LDL C) treatment goal of < 100 mg/dL and the optional LDL-C target of < 70 mg/dL with coadministration of ezetimibe and simvastatin (EZE/SIMVA) vs either atorvastatin or simvastatin monotherapy. PATIENTS: Patients with established CHD or CHD risk equivalent according to National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) criteria with baseline LDL-C = 130 mg/dL and triglycerides (TG) < or = 350 mg/dL. METHODS: A post hoc analysis from 2 separate studies assessed the percentage of high-risk patients achieving the LDL-C targets (< 100 and < 70 mg/dL) after 6 weeks on the usual recommended starting doses of the following treatments: EZE/SIMVA (10/20 mg) vs atorvastatin (10 mg) or simvastatin (20 mg). Depending on the study, EZE/SIMVA 10/10 or 10/40 mg was also compared with either atorvastatin 10 mg or simvastatin 20 mg. Percent change in other lipid parameters from baseline to study endpoint was also examined. RESULTS: In both studies, the proportions of patients achieving an LDL-C of < 100 mg/dL were significantly (P < .001) greater for EZE/SIMVA 10/10, 10/20, or 10/40 mg vs either atorvastatin 10 mg or simvastatin 20 mg after 6 weeks. The percentage reaching the optional LDL-C treatment target of < 70 mg/dL was also significantly higher with EZE/SIMVA compared with either atorvastatin or simvastatin. Percent reduction in LDL-C was significantly (P < .001) larger with all doses of EZE/SIMVA (46% to 59%) compared with either atorvastatin 10 mg (37%) or simvastatin 20 mg (38%) monotherapy after 6 weeks. Changes in other lipid parameters consistently favored EZE/SIMVA vs statin monotherapy. All treatments were well tolerated in both studies. CONCLUSION: Patients at high risk for CHD are more likely to attain LDL-C treatment targets with the usual recommended starting dose of EZE/SIMVA (10 or 20 mg) therapy than with that of atorvastatin (10 mg) or simvastatin (20 mg) monotherapy. PMID- 16369231 TI - Diagnosis and management of injuries from dangerous marine life. AB - Injuries from marine life encompass a wide spectrum, from mild stings to severe bites. Fortunately most of the injuries are mild, although some may be significant, resulting in death. Most of these injuries can be treated by family physicians with a knowledge of the cause of the pathology. Over the years, there have been many treatment options. Some have actually caused an increase in severity. An important rule in treating these injuries is to inactivate the venom, treat the local reaction or injury, and treat the systemic sequelae. Jellyfish stings are the most common type of marine injury. The tentacles possess nematocysts, which are stinging units that are inactivated by the application of vinegar. Sea urchin and stingray injuries require the removal of the imbedded spines after the wound is soaked in hot water. Coral, sea bathers eruption, and swimmer's itch require thorough scrubbing and irrigation. Sea snakes, cone shells, and venomous fish possess a neurotoxin that requires close monitoring in the event of cardiopulmonary collapse. All of these injuries require tetanus status monitoring and consideration of coverage for infectious sequelae. PMID- 16369232 TI - Coercive restraint therapies: a dangerous alternative mental health intervention. AB - Physicians caring for adopted or foster children should be aware of the use of coercive restraint therapy (CRT) practices by parents and mental health practitioners. CRT is defined as a mental health intervention involving physical restraint and is used in adoptive or foster families with the intention of increasing emotional attachment to parents. Coercive restraint therapy parenting (CRTP) is a set of child care practices adjuvant to CRT. CRT and CRTP have been associated with child deaths and poor growth. Examination of the CRT literature shows a conflict with accepted practice, an unusual theoretic basis, and an absence of empirical support. Nevertheless, CRT appears to be increasing in popularity. This article discusses possible reasons for the increase, and offers suggestions for professional responses to the CRT problem. PMID- 16369233 TI - Disseminated aspergillosis following infliximab therapy in an immunosuppressed patient with Crohn's disease and chronic hepatitis C: a case study and review of the literature. AB - A 55-year-old white woman with a greater than 25-year history of Crohn's disease developed disseminated aspergillosis following combination therapy with Methylprednisolone, azathioprine, and infliximab. The patient was hospitalized 11 days after initiation of infliximab for respiratory symptoms and developed respiratory failure, coma, and died. Postmortem examination revealed disseminated Aspergillus fumigatus involving multiple organs. This case demonstrates that combined treatment with infliximab, methylprednisone, and azathioprine may induce severe immunosuppression and depressed cellular immunity, leading to severe opportunistic infections. Given the increasing use of antitumor necrosis factor agents, physicians should be aware of the risk of opportunistic infections and be vigilant about diagnosing and aggressively treating these infections to reduce the risk of disseminated disease. PMID- 16369234 TI - Bilateral radial head fractures in a woman with trivial trauma. AB - Radial head fractures are common injuries, occurring in about 20% of all acute elbow injuries. Isolated radial head fractures are not common and include about 2% of all fractures around the elbow. Bilateral radial head fractures are rare and usually associated with severe trauma and associated fractures and dislocations. We report a case of bilateral undisplaced radial head fracture in a woman, following a simple fall. Early recognition, proper management, and physical therapy led to complete recovery and full functional movement of the elbow. PMID- 16369235 TI - Severe hepatotoxicity in a rheumatoid arthritis patient switched from leflunomide to methotrexate. PMID- 16369236 TI - Case report: psoas abscess--unusual etiology of groin pain. PMID- 16369237 TI - The 2-bodied continuous cardiac output catheter. AB - Continuous cardiac output (CCO) pulmonary artery catheters (PACs) are used in cardiac surgical patients for hemodynamic monitoring. This is an invasive technique; therefore, it has well-known mechanical and infectious complications related to its use. Associated problems, more commonly noticed, are catheter malfunctions and malpositions. We present a case in which a CCO catheter appeared to have a double body on radiologic diagnosis. This resulted in extra radiography for rechecking and careful clinical reevaluation of the monitored data. PMID- 16369238 TI - When is a difference not a difference: medicine or shooting craps? PMID- 16369239 TI - Health at every size: toward a new paradigm of weight and health. PMID- 16369240 TI - Healthcare professionals and dual loyalty: technical proficiency is not enough. PMID- 16369241 TI - Ileocolonic intussusception. PMID- 16369242 TI - Hereditary diffuse gastric cancer. PMID- 16369244 TI - A 2-year-old girl with persistent ascites. PMID- 16369243 TI - Irritable bowel syndrome and functional dyspepsia: different diseases or a single disorder with different manifestations? PMID- 16369245 TI - Probiotics in the management of inflammatory bowel disease. PMID- 16369246 TI - Benefit with aromatase inhibitors in the adjuvant setting for postmenopausal women with breast cancer. AB - The third-generation aromatase inhibitors, letrozole, anastrozole, and exemestane, have been shown to be effective both as alternatives to tamoxifen in first-line treatment of hormone-sensitive advanced breast cancer in postmenopausal women and following failure of first-line tamoxifen for endocrine therapy. These 3 agents are now being investigated as adjuvant therapy of early breast cancer, as alternative or complementary treatments to the standard, tamoxifen. Three treatment strategies are under investigation: replacement of tamoxifen as adjuvant therapy for 5 years (early adjuvant therapy), sequencing of tamoxifen before or after an aromatase inhibitor during the first 5 years (early sequential adjuvant therapy), or following 5 years of tamoxifen (extended adjuvant therapy). In the first adjuvant trial (Arimidex, Tamoxifen Alone or in Combination [ATAC]), anastrozole was significantly superior to tamoxifen in reducing risk of disease recurrence, and recently, the Breast International Group (BIG) trial BIG 1-98 demonstrated the significant superiority of letrozole over tamoxifen in improving disease-free survival. A large trial (International Collaborative Cancer Group [ICCG] trial 96) investigated sequencing of 2 to 3 years of exemestane after 2 to 3 years of tamoxifen and found that switching to exemestane was significantly superior in disease-free survival compared with continuing on tamoxifen. The Arimidex or Nolvadex (ARNO) and the small ITA (Italian Tamoxifen Arimidex) trials similarly sequenced anastrozole after tamoxifen and also found that sequencing reduced the hazard of recurrence compared with remaining on tamoxifen. Trial MA.17 evaluated extended adjuvant therapy with letrozole vs placebo following 5 years of tamoxifen. Disease-free survival was significantly improved with letrozole vs placebo, irrespective of whether patients had lymph node-positive or node-negative tumors. All 3 aromatase inhibitors were generally well tolerated. Results of these trials indicate that aromatase inhibitors provide important benefits relative to tamoxifen in each of these adjuvant treatment settings, but the optimal approach still needs to be defined. Other trials continue to investigate some of these adjuvant treatment strategies. PMID- 16369247 TI - Does the use of an automated tool for self-reporting mood by patients with bipolar disorder bias the collected data? AB - CONTEXT: Automating data collection from patients can improve data quality, enhance compliance, and decrease costs in longitudinal studies. About half of all households in industrialized countries now have a home computer. OBJECTIVE: While we previously validated the ChronoRecord software for self-reporting mood on a home computer with patients who have bipolar disorder, this study further investigates whether this technology created a bias in the collected data. METHODS: During the validation study, 80 of 96 (83%) patients returned 8662 days of data (mean, 114.7 +/- 32.3 SD days). The patients' demographics were compared with those of similar longitudinal studies in which patients used paper-based data collection tools. In addition, because demographic characteristics may influence attitudes toward technology, observer-rated scores on the Hamilton Depression Rating Scale and Young Mania Rating Scale were used to group patients by severity of illness, and the self-reported mood ratings were analyzed for evidence of bias from the patients' gender, ethnicity, diagnosis, age, disability status, or years of education. Analysis was performed using the 2-way analysis of variance and general linear model. RESULTS: The patients' demographic characteristics were very similar to those of patients with bipolar disorder who participated in comparable longitudinal studies using paper-based tools. After grouping the patients by severity of illness, none of the demographic variables had a significant effect on the patients' self-reported mood using the automated tool. CONCLUSION: The use of a computer does not seem to bias sample data. As with studies using paper-based self-reporting, results from studies of patients using ChronoRecord software on a home computer to report mood can be generalized. PMID- 16369248 TI - Venlafaxine and serious withdrawal symptoms: warning to drivers. AB - Venlafaxine is a widely used serotonin- and norepinephrine-reuptake inhibitor type antidepressant that causes serious adverse effects in at least 5% of cases. Serious withdrawal symptoms may occur within hours of cessation or reduction of the usual dosage and may affect motor and coordination skills to such a degree that patients should be explicitly urged either to adhere to a strict medication routine or not to drive a car. Recent clinical evidence about withdrawal symptoms is presented that may indicate incidents in noradrenergic activity irrespective of dosage. PMID- 16369250 TI - Reader's response to "medical investigations of homicides of prisoners of war in Iraq and Afghanistan". PMID- 16369251 TI - "Aldosterone escape" or refractory hyperaldosteronism? PMID- 16369252 TI - Readers' responses to "why should physicians be underlings?". PMID- 16369254 TI - Readers' responses to "open access medical publishing is finally coming alive". PMID- 16369255 TI - Readers' responses to "boxing should be banned in civilized countries--round 4". PMID- 16369257 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16369259 TI - Reader's response to "we can cure most breast cancers with early diagnosis and treatment". PMID- 16369260 TI - Readers' responses to "helping families cope with sudden or unexpected death". PMID- 16369262 TI - Readers' responses to "breaking the rules: entertainment and public health". PMID- 16369263 TI - Reader's response to "kids and media: learning happens". PMID- 16369264 TI - Readers' responses to "new Medicare rule forces cancer, heart, and lung patients into nursing homes". PMID- 16369265 TI - Reader's response to "the importance of measuring quality and performance in healthcare". PMID- 16369267 TI - Readers' responses to "the real world of medical practice?". PMID- 16369268 TI - Why is consumer-directed health care (CDHC) growing rapidly in America? PMID- 16369269 TI - How can we align the incentives to sustain shared electronic health records in communities? PMID- 16369270 TI - Did your genes give you that sore throat? PMID- 16369271 TI - Taking kidneys from children for transplants cannot be justified. PMID- 16369272 TI - Are 80 hours a week enough to train a surgeon? PMID- 16369273 TI - It is time for marijuana to be reclassified as something other than a Schedule I drug. PMID- 16369274 TI - Thought for food: 2400-year-old advice for today's worldwide weight crisis. PMID- 16369275 TI - The crisis in America's laboratories: it's every physician's business. PMID- 16369276 TI - Physicians must participate in the quality movement. PMID- 16369277 TI - Doctors and patients working together. PMID- 16369278 TI - Boxing should be banned in civilized countries--round 4. PMID- 16369279 TI - We must not lose the wonders of vaccination. PMID- 16369280 TI - Why should physicians be underlings? PMID- 16369281 TI - Open access medical publishing is finally coming alive. PMID- 16369282 TI - In the wake of Katrina: a surgeon's first-hand report of the New Orleans tragedy. PMID- 16369283 TI - Institute for Quality in Laboratory Medicine: Recognizing Excellence in Practice: highlights from first landmark summit--an opportunity to enhance medical care; April 28-30, 2005; Atlanta, Georgia. PMID- 16369284 TI - Case 10: it's "the vision thing". PMID- 16369285 TI - Omissions, biases, and nondisclosed conflicts of interest: is there a hidden agenda in the NAMS position statement? PMID- 16369286 TI - Prevention of ovarian hyperstimulation syndrome by early aspiration of small follicles in hyper-responsive patients with polycystic ovaries during assisted reproductive treatment cycles. AB - OBJECTIVE: This study was designed to assess the efficacy of early aspiration of small follicles (EASF) in preventing moderate and severe ovarian hyperstimulation syndrome (OHSS) in women with polycystic ovaries who experienced excessive ovarian response during the early part of assisted reproductive treatment cycles (ART). STUDY DESIGN: EASF was undertaken transvaginally during 15 in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) cycles in 14 women with polycystic ovaries who produced more than 20 small follicles during the early part of the cycle. A minimum of 10 small follicles. OUTCOMES: None of the patients developed moderate or severe OHSS. There were 9 intrauterine pregnancies (60 %) and 1 ectopic pregnancy. CONCLUSIONS: EASF 4 to 7 days after starting gonadotropin injections prevented moderate and severe OHSS in susceptible patients with polycystic ovaries and resulted in a high pregnancy rate. Larger studies are needed to verify these promising results. PMID- 16369287 TI - Diagnosis of labor: a prospective study. AB - Accurate diagnosis of the onset of labor remains a problem in obstetrics. Criteria commonly used to diagnose labor have never been scientifically evaluated. This prospective study involved 423 pregnant women who presented themselves with uterine contractions to 2 Italian hospitals (248 nulliparous patients total and 175 multiparous total) and who were either admitted or advised to return home. The obstetrician on duty collected data using a standardized form that listed common criteria for labor diagnosis. Multivariate analysis showed that a reduction of the interval between consecutive uterine contractions, odds ratio [OR] = 1.42; 95% confidence interval [95%CI] 1.06-1.90); abdominal pain of increasing intensity (OR = 1.42; 95% CI 1.01-2.02); cervical effacement (OR = 1.4; 95%CI 1.12-1.77); and cervical dilation (OR = 1.91; 95% CI 1.53-2.38) were significant markers of the onset of labor. On the other hand, backache had a negative diagnostic value (OR = 0.78; 95% CI 0.61-0.99). The value of criteria such as regular uterine contractions, loss of mucous plug, changes in intestinal habits, vomiting, pain that is relieved by walking, and changes in breathing pattern did not reach statistical significance. PMID- 16369288 TI - Addressing the need for a simpler algorithm for the management of women with urinary incontinence. AB - AIMS: Urinary incontinence (UI) is a prevalent and bothersome condition for many women. However, few women convey the problem to primary care physicians (PCPs), and few PCPs proactively ask about it. One reason may be that available UI recommendations are too complex and difficult to implement. Simplification may encourage greater adaptation as long as a simplified algorithm does not compromise patient safety. I review the opportunity to develop a simpler UI algorithm based on the 3rd edition 2005 International Consultation of Incontinence (ICI) guidelines for the initial management of UI in women. METHODS: A literature search for scientific evidence supporting simplification was performed using PubMed, the Cochrane Database, the Agency for Health Care Policy and Research's (AHCPR) 1996 recommendations, and the ICI's 2005 Incontinence Book / CD-ROM. Each level in the ICI algorithm for initial assessment and treatment was evaluated to develop an updated and less complex management strategy. RESULTS: A simpler UI algorithm for PCPs should focus on considering reversible risk factors and identifying those who need specialized management. A short questionnaire may be sufficient to assess type and severity of UI, whereas a standard neurologic or comprehensive abdominal and pelvic examination seems unnecessary. Treatment should be restricted to a few evidence-based options. CONCLUSIONS: In conclusion, a simpler UI algorithm allows the initiation of evidence-based treatment after a simplified assessment in primary care. PMID- 16369289 TI - A 26-year-old woman presented to Kijabe Mission Hospital with a diffuse rash and dyspnea. PMID- 16369291 TI - Hypertriglyceridemia in antiretroviral therapy. PMID- 16369292 TI - Disease progression among untreated HIV-infected patients in South Ethiopia: implications for patient care. AB - CONTEXT: The natural course of HIV disease progression among resource-poor patient populations has not been clearly defined. OBJECTIVE: To describe predictors of HIV disease progression as seen at an outpatient clinic in a resource-limited setting in rural Ethiopia. DESIGN: This prospective cohort study included all adult HIV patients who visited an outpatient clinic at Arba Minch hospital in South Ethiopia between January 30, 2003 and April 1, 2004. Clinical and hematologic measurements were done at baseline and every 12 weeks thereafter until the patient was transferred, put on antiretroviral therapy, was lost to follow-up, or died. Community agents reported patient status every month. SETTING: A district hospital with basic facilities for HIV testing and patient monitoring. MAIN OUTCOME MEASURES: Death, diagnosis of tuberculosis, and change in disease stage. RESULTS: We followed 207 patients for a median duration of 19 weeks (range, 0-60 weeks). A total of 132 (64%) of them were in WHO stage III. The overall mortality rate was 46 per 100 person-years of observation (PYO). Mortality increased with advancing disease stage. Diarrhea, oral thrush, and low total lymphocyte count were significant markers of mortality. The incidence of tuberculosis was 9.9 per 100 PYO. Baseline history of easy fatigability and fever were strongly associated with subsequent development of tuberculosis. CONCLUSIONS: The mortality rate and the incidence of tuberculosis in our cohort are among the highest ever reported in sub-Saharan Africa. We identified oral thrush, diarrhea, and total lymphocyte count as predictors of mortality, and easy fatigability and fever as predictors of tuberculosis. The findings have practical implications for patient care in resource-limited settings. PMID- 16369293 TI - Initiating antiretrovirals in a resource-constrained setting: does one size fit all? PMID- 16369294 TI - Frequency of drug-resistant variants of HIV-1 coexistent with wild-type in treatment-naive patients of India. AB - CONTEXT: Over the past few years, reports of emergence and transmission of drug resistant strains of HIV have increased, especially in western countries. In the context of increased widespread use of zidovudine- and lamivudine-based combinations in India, coupled with the genetic diversity of HIV, it is essential to generate preliminary data on the frequency of zidovudine- and lamivudine resistant variants of HIV-1 in North India. OBJECTIVES: In the present study, the authors screened for mutations in the pol gene of HIV-1 associated with resistance to zidovudine and lamivudine in HIV-infected treatment-naive patients from North India. DESIGN AND PATIENTS: The mutations were screened at codons 70 and 215 (conferring resistance to zidovudine) and at codon 184 (conferring resistance to lamivudine) by using a nested amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) approach from the proviral DNA of 60 patients. RESULTS: Most of the patients showed a mixture of both wild-type and mutant virus. In all but 1 patient, wild-type virus was observed with respect to each codon. Mutant variants were also observed in many patients, especially at codon 70 (48 patients [80%]) and codon 184 (19 patients [31.67%]). In contrast, the frequency of mutation at codon 215 was found to be very low (1 patient [1.67%]). CONCLUSIONS: In this sample of treatment-naive HIV-1-infected patients in North India, a high proportion of mutant variants harbored mutations in the pol gene at codons- 70 and 184 coexisting with wild-type HIV-1. PMID- 16369295 TI - Genetic barriers to resistance and impact on clinical response. AB - The development of drug resistance and cross-resistance continues to pose a challenge to successful long-term antiretroviral therapy despite the availability of new antiretroviral agents. The genetic barrier to resistance of a regimen does not directly correlate with its effectiveness. For some regimens with a low genetic barrier to resistance, however, the emergence of only 1 or 2 key resistance mutations may confer drug resistance not only to that regimen but also to other agents, thereby limiting subsequent treatment options. In addition to the genetic barrier to resistance, factors such as efficacy, safety, tolerability, convenience, and adherence must be considered when choosing a regimen. PMID- 16369297 TI - Diagnosis, treatment, and management of irritable bowel syndrome with constipation and chronic constipation. AB - Irritable bowel syndrome with constipation (IBS-C) and chronic constipation are 2 common gastrointestinal motility disorders that place a substantial burden on patients and society. Symptoms of both disorders are chronic, sometimes severe, and often respond poorly to treatment with traditional approaches, resulting in reduced quality of life, polypharmacy, and frequent healthcare utilization. Because structural, physical, or biochemical markers cannot be used to identify either disorder, diagnosis is symptom-based. In the absence of alarm features suggestive of organic disease or secondary causes of symptoms, these disorders can be positively and confidently diagnosed. In general, traditional agents used to treat patients with constipation target only a single symptom, and do not provide adequate relief of symptoms in the majority of IBS-C patients. Although patients with mild constipation symptoms may respond to treatment with fiber and laxatives, others with moderate-to-severe symptoms may require additional therapies and/or referral to a specialist for further evaluation. The advent of novel serotonergic agents has rejuvenated the therapeutic approach to patients with IBS-C and chronic constipation. PMID- 16369298 TI - Robotic surgery: applications, limitations, and impact on surgical education. PMID- 16369299 TI - Robotic surgery: applications, limitations, and impact on surgical education. PMID- 16369300 TI - Religion, health, and questions of meaning. PMID- 16369301 TI - The healing potential of hospital food. PMID- 16369302 TI - Aspirin resistance and its implications in clinical practice. AB - Recent studies have suggested that some patients may not obtain the full benefits of aspirin's antiplatelet effects. An international roundtable of experts was held November 6, 2004, in New Orleans, Louisiana, to address the concept of aspirin resistance and its clinical implications. Panelists discussed various definitions and possible mechanisms of aspirin resistance, along with several tests currently available to measure platelet function. They recommended that until the clinical importance of these tests is known, physicians should continue to prescribe low-dose aspirin therapy for individuals at high risk for cardiovascular events and patients currently taking aspirin should continue to comply with their prescribed therapy. PMID- 16369303 TI - Obesity: pain and prejudice. PMID- 16369304 TI - Uncle Sam's diet sensation: MyPyramid--an overview and commentary. AB - MyPyramid symbolizes the federal nutrition policy, the 2005 Dietary Guidelines for Americans, replacing the 1992 Food Guide Pyramid. This Internet-based educational tool, available at http://MyPyramid.gov , provides general guidance for food selection from 5 food groups (grains, vegetables, fruit, milk, meat and beans). Additionally, MyPyramid addresses the obesity crisis by placing a strong emphasis on reducing calorie consumption and increasing physical activity. Although some experts praise this new, interactive pyramid, many believe that the government missed an opportunity to direct Americans toward healthier diets. Furthermore, the information available at MyPyramid.gov is not likely to be accessed by the populations that need it most: the underprivileged. Because obesity prevention programs structured only to educate and inform have generally been unsuccessful, the federal government must take the next step and revise agriculture policies and programs to promote healthy eating. PMID- 16369306 TI - Pregnancy and other factors associated with higher CD4+ T-cell counts at HIV diagnosis in Southeast Michigan, 1992-2002. AB - BACKGROUND: Early diagnosis of HIV infection provides the opportunity for treatment to prevent progression to AIDS and for intervention to prevent further transmission. The impact of routine screening of pregnant women and other factors on the stage of HIV disease at diagnosis were evaluated. METHODS: Data were collected in 1992-2002 from the medical records of persons presenting for HIV related care at 2 major medical centers in Detroit, Michigan. Patients were included in the analysis if they had a CD4+ T-cell count recorded within 6 months of their first positive HIV test (N = 1858). RESULTS: Half of the patients (49%) had a first CD4+ T-cell count of < 200 cells/mm3 and 19% had an AIDS-defining illness at the time of HIV diagnosis. In the multivariate model, pregnant women were less likely than nonpregnant women to enter care with a CD4+ T-cell count of < 200 cells/mm3 (odds ratio, .24; 95% confidence interval, .14-.41). Even after adjusting for pregnancy, female sex was protective, as was age < 30 years. HIV transmission risk factors, race, and time period of HIV diagnosis were not significantly associated with first CD4+ T-cell counts of < 200 cells/mm3. CONCLUSION: Routinely offering HIV testing in prenatal care, as required by Michigan law, resulted in earlier diagnoses of HIV in pregnant women, as indicated by their higher CD4+ T-cell counts. Increasing routine HIV testing of all persons seeking medical care may increase the overall proportion of HIV diagnoses that are made early in the disease process. PMID- 16369307 TI - Medical students' self-reported typical counseling practices are similar to those assessed with standardized patients. AB - CONTEXT/OBJECTIVES: We surveyed fourth-year medical students in the Class of 2003 at Emory University School of Medicine, Atlanta, Georgia, about various personal and clinical practices. We were especially interested in the frequency that these seniors reported of talking with patients about nutrition, weight, exercise, alcohol, and cigarette smoking. Because the validity of our findings about these counseling practices was limited by our having only self-reported data from seniors' questionnaires, we developed a standardized patient (SP) examination to test the relationship between what students reported on the questionnaires and how they actually performed with SPs. DESIGN/SETTING/MAIN OUTCOME MEASURES: As part of a lengthy questionnaire, 88 senior medical students answered these 5 separated questions: "With a typical general medicine patient, how often do you actually talk to patients about: (1) nutrition; (2) exercise/physical activity; (3) weight; (4) smoking cessation (among smokers); and (5) alcohol? (never/rarely, sometimes, usually/always)." As part of their internal medicine subinternship final exam, students clinically assessed 4 SP cases with predetermined risk factors (poor diet, exercise, alcohol, and/or cigarette smoking habits). RESULTS: For every risk factor, the proportion of SPs actually counseled was higher for those students who self-reported discussing that risk factor more frequently with their patients. Additionally, the odds of counseling an SP for any risk factor were significantly higher (odds ratio = 1.76-2.80, P < .05) when students reported more frequent counseling. CONCLUSION: Student self reports regarding patient counseling may be useful when resources are limited, and the purpose is to grossly and anonymously distinguish between higher and lower performers. PMID- 16369308 TI - Diet and diabetic retinopathy: insights from the Diabetes Control and Complications Trial (DCCT). AB - OBJECTIVE: We explore the influence of lifestyle on the progression of retinopathy. DESIGN: Post hoc statistical analysis. SUBJECTS/SETTING: One thousand forty-one patients with type 1 diabetes from 29 specialty clinics. INTERVENTION: The Diabetes Control and Complications Trial (DCCT) lifestyle data (diet, exercise, and tobacco use) and retinopathy-related risk factors (mean arterial pressure, the low-density lipoprotein/high-density lipoprotein cholesterol ratio [LDL-C/HDL-C], serum triglycerides, glycosolated hemoglobin [HbA1c] levels, body mass index [BMI], and insulin utilization) were related to the rate of progression of retinopathy. MAIN OUTCOME MEASURES: Correlation between lifestyle data with progression of retinopathy and retinopathy-related risk factors. RESULTS: The percentage of calories as total fatty acids at baseline and overall positively correlated with prestudy and overall progression of retinopathy (r = .15, P < .0001 and r = .14, P < .0001, respectively). Average overall percentage of calories as dietary fiber inversely correlated with prestudy and overall progression of retinopathy (r = -.07, P = .0102 and r = .10, P < .0002, respectively). The progression of retinopathy correlated with mean arterial pressure (prestudy r = .09, P = .0004 and overall r = .20, P < .0001), LDL-C/HDL-C (prestudy r = .13, P < .0001 and overall r = .15, P < .0001), serum triglycerides (prestudy r = .18, P < .0001 and overall r = .26, P < .0001), HbA1c (prestudy r = .10, P < .0001 and overall r = .45, P < .0001), BMI (prestudy r = .08, P <.0034 and overall r = .05, P = .08), insulin utilization (prestudy r = .19, P < .0001 and overall r = .14, P < .0001), tobacco use (prestudy r = .08, P < .0231 and overall r = .09, P < .0011), and the intensive vs conventional therapy study group (on-study r = -.27, P < .0001). CONCLUSION: Tobacco use and diet, particularly the consumption of fatty acids and dietary fiber, are significantly associated with the rate of progression of diabetic retinopathy and retinopathy-related risk factors. PMID- 16369309 TI - Strategies to enhance patient adherence: making it simple. AB - The problem of poor patient adherence has been extensively researched, but the rates of nonadherence have not changed much in the past 3 decades. Healthcare providers play a unique and important role in assisting patients' healthy behavior changes. We conducted a narrative review of the current literature to help providers become more familiar with proven interventions that can enhance patient adherence. We then grouped the interventions into categories that can be remembered by the mnemonic "SIMPLE":1. Simplifying regimen characteristics; 2. Imparting knowledge; 3. Modifying patient beliefs; 4. Leaving the bias; and 6. Evaluating adherence. Chronic lifestyle behavior change often requires a combination of all the aforementioned strategies. We suggest a conceptual framework, which calls for a multidisciplinary approach with the above strategies in the context of a healthcare team and system-related factors. We hope that this framework would not only help design scientifically proven interventions, but also reduce the time and cost involved with implementing these strategies in a healthcare setting. PMID- 16369310 TI - Proactive interference, retroactive interference--what about self-interference? A new interpretation of the recency-primacy shift. AB - The recency-primacy shift (RPS) indicates that memory for early list items improves and memory for later items becomes worse as the retention interval between study and test increases. In this contribution, this puzzling experimental finding--memory improving with time--is found to be consistent with a model in which recognition is temporarily interfered with by its own storage process (self-interference). I show that this interpretation can qualitatively better account for the RPS experimental data than can the dimensional distinctiveness model, the only other outstanding explanation of the RPS. Two experimental predictions separate the 2 models: The dimensional distinctiveness model predicts no RPS for 2-item lists, in contrast to self-interference, and as the overall timescale is changed, the dimensional distinctiveness model predicts no difference in the RPS whereas self-interference predicts significant changes. PMID- 16369311 TI - Escitalopram in adolescent major depression. AB - Escitalopram is the purified functional isomer contained in citalopram. Escitalopram is now prescribed in 26 countries. In the United States, the only US Food and Drug Administration (FDA)-approved selective serotonin reuptake inhibiter (SSRI) for adolescents is fluoxetine. However, in clinical practice all antidepressants are used in adolescents. Five patients had parents who opted for the use of escitalopram instead of other treatments. Reasons included poor response and side effects from other SSRIs. Specifically, escitalopram was considered possibly less likely to cause obesity than paroxetine. It also caused a lower frequency of akathisia than fluoxetine, more stable blood levels over years than sertraline, very low drug interactions, and a low onset of anxiety if using a 5-mg starting dose. Although studies in adolescents are very limited for escitalopram, its parent medication--citalopram--has been used in over 40 million patients. Parents and adolescent patients should be made aware of all antidepressant options, if psychopharmacology is indicated. In some patients, escitalopram may have use. PMID- 16369312 TI - A case of sarcoidosis in a patient with IgA nephropathy. PMID- 16369313 TI - Hepatic dysfunction in a patient with Plasmodium vivax infection. PMID- 16369314 TI - Antifibrinolytic agents in traumatic hemorrhage. PMID- 16369315 TI - Searching in the darkness: about prayer and medical cures. PMID- 16369316 TI - Standardization vs diversity: how can we push peer review research forward? PMID- 16369317 TI - Gastrointestinal bleeding from an obscure cause. PMID- 16369318 TI - A 2-month-old with persistent diarrhea. PMID- 16369319 TI - Acute dysphagia after carp fish meal: a case report. AB - An 80-year-old man was admitted to the emergency department with acute dysphagia symptoms of 8 hours' duration. The problem began after a carp fish meal. Prominent symptoms included inability to swallow, mild dyspnea, and cough. Laboratory evaluation and fiber-optic examination of the upper aerodigestive tract was normal except for accumulation of saliva in the hypopharynx. Examination with a flexible esophagoscope revealed a total blockage of the lumen of the lower esophagus due to impaction of carp fish roe (column-like formation). With careful maneuvers and water pressure we were able to relieve the obstruction. No other lesion was detectable and no further interventions were needed. Patient follow-up was uneventful. PMID- 16369320 TI - Chronic pancreatic pain. PMID- 16369321 TI - Lipoma case report. PMID- 16369322 TI - Expert commentary--new developments in the treatment of constipation. PMID- 16369324 TI - Topics in pediatric leukemia--hematopoietic stem cell transplantation. PMID- 16369323 TI - Expert commentary--bloating, distension, and the irritable bowel syndrome. PMID- 16369325 TI - Topics in pediatric leukemia--acute myeloid leukemia. PMID- 16369326 TI - Topics in pediatric leukemia--myelodysplastic and myeloproliferative disorders of childhood. PMID- 16369327 TI - Topics in pediatric leukemia--acute lymphoblastic leukemia and late effects in long-term survivors. PMID- 16369328 TI - Topics in pediatric leukemia--acute lymphoblastic leukemia. PMID- 16369329 TI - Time and the biology of aging. PMID- 16369330 TI - Phytoestrogens and antioxidants--bits of experimental evidence. PMID- 16369331 TI - Brain metastasis from cervical carcinoma--a case report. AB - Brain metastases from cervical carcinomas are extremely rare. We report a patient with squamous cell carcinoma of the cervix who developed an isolated left parietooccipital lobe metastasis within 4 months of treatment of the primary disease. The presenting symptoms of the metastatic disease were visual disturbance, headache, and vomiting. The patient was successfully treated by surgical excision of the metastasis and adjuvant whole brain radiation therapy, and she was disease-free at the 6-month follow-up after treatment of the recurrence. PMID- 16369332 TI - Omalizumab: a monoclonal anti-IgE antibody. AB - OBJECTIVES: To describe allergic asthma and allergic rhinitis pathophysiology and review the pharmacologic, pharmacokinetic, pharmacodynamic, efficacy, and safety data for omalizumab. METHODS: MEDLINE, In-Process & Other Non-Indexed Citations, and EMBASE Drugs & Pharmacology were searched using olizumab, omalizumab, E25, rhuMAb-E25, and anti-IgE. Combinations of rhinitis, asthma, and IgE captured information on disease pathophysiology. RESULTS: Omalizumab is a monoclonal antibody targeting the high-affinity receptor binding site on human immunoglobulin (Ig)E. Bound IgE is not available for basophil binding, degranulation is attenuated, and allergic symptoms are reduced. In asthma trials, omalizumab reduced inhaled corticosteroid and rescue medication requirements and improved asthma control and asthma quality of life in moderate to severe allergic asthmatics with disease poorly controlled by inhaled corticosteroids. In trials of patients with poorly controlled moderate to severe seasonal allergic rhinitis (SAR), omalizumab reduced the severity of exacerbations and rescue medication use, and improved rhinitis-related quality of life. Benefits were also observed in trials utilizing combinations of immunotherapy and omalizumab for SAR and in trials of perennial allergic rhinitis (PAR). Omalizumab has been well tolerated. Although malignant neoplasms have been observed in treated patients, they were likely not related to omalizumab therapy. CONCLUSIONS: Omalizumab has demonstrated efficacy in children, adults, and adolescents with uncontrolled moderate to severe allergic asthma and allergic rhinitis. Long-term safety beyond 52 weeks needs continued evaluation. PMID- 16369335 TI - More serious letters about obesity. PMID- 16369334 TI - Neuropathic arthropathy of the shoulder (Charcot shoulder). PMID- 16369336 TI - Readers' responses to the webcast video editorials entitled "the American healthcare 'system' in 2005"--parts 1 and 2. PMID- 16369337 TI - Readers' responses to the webcast video editorial entitled "caution--lab test ordering trouble ahead". PMID- 16369338 TI - Readers' responses to the webcast video editorial entitled "evidence-based medicine or faith-based medicine?". PMID- 16369339 TI - Reader's response to the webcast video editorial entitled "the institute for one world health--a not-for-profit pharmaceutical company". PMID- 16369340 TI - Readers' responses to the webcast video editorial entitled "Willie Sutton and screening for alcohol in the emergency department and trauma center". PMID- 16369342 TI - Reader's response to "increasing orgasm and decreasing dyspareunia by a manual physical therapy technique". PMID- 16369343 TI - Readers' responses to the webcast video editorial entitled "the official ethics of the American medical profession". PMID- 16369344 TI - Reader's response to the webcast video editorial entitled "is chronic alcoholism another pediatric disease?". PMID- 16369345 TI - Readers' positive responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16369346 TI - Readers' negative or neutral responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16369347 TI - reader's response to the webcast video editorial entitled "the international code of medical ethics of the World Medical Association". PMID- 16369348 TI - Reader's response to "advances in diabetes for the millennium: an e-symposium". PMID- 16369351 TI - Reader's response to the article entitled "a recent exposure to mustard gas in the United States: clinical findings of a cohort (n = 247) 6 years after exposure". PMID- 16369352 TI - Impact of health savings accounts on the road map to universal healthcare. PMID- 16369354 TI - Readers' responses to the article entitled "anticoagulation therapy for venous thromboembolism". PMID- 16369355 TI - How to control healthcare costs: take a walk on the supply side. PMID- 16369356 TI - The American healthcare "system" in 2005--part 7: a public-private mix is the best way to fix our broken system. PMID- 16369357 TI - The American healthcare "system" in 2005--part 6: how to grade the current system and proposed reforms. PMID- 16369358 TI - How to take useful drugs without hurting yourself. PMID- 16369359 TI - The American healthcare "system" in 2005--part 5: 2 good options for fixing our broken system. PMID- 16369360 TI - The American healthcare "system" in 2005--part 4: why not the single-payer solution? PMID- 16369361 TI - The American healthcare "system" in 2005--part 3: why not put the consumer in charge? PMID- 16369362 TI - How to prevent obesity in developed countries--part 2. PMID- 16369363 TI - The American healthcare "system" in 2005--part 2: who is in charge? PMID- 16369364 TI - The American healthcare "system" in 2005--part 1: context. PMID- 16369365 TI - Caution--lab test ordering trouble ahead. PMID- 16369366 TI - The Institute for One World Health--a not-for-profit pharmaceutical company. PMID- 16369367 TI - A doctor's duty in a Palestinian prison camp in Israel. PMID- 16369368 TI - Outcome predictors in nonoperative management of newly diagnosed subacromial impingement syndrome: a longitudinal study. AB - OBJECTIVE: This prospective investigation is designed to determine the prognostic factors associated with the response to conservative therapy of subacromial impingement syndrome. MATERIALS AND METHODS: We treated 102 patients with subacromial impingement syndrome, diagnosed by physical examination and a subacromial lidocaine injection test, with a standardized conservative protocol. We followed the patients for a period of 12 months. Outcome was evaluated with Constant score and effects of 8 variables: Age, sex, pretreatment symptom duration, dominant shoulder, initial Constant score, active range of motion, acromion morphology, and acromial spur on patient outcomes were evaluated with univariate and multivariate analyses. RESULTS: Among 89 patients (44 men; mean age, 56.4 years) who finished the study, the mean difference between initial and final scores was 15.9 (95% confidence interval [CI]: 13.9-17.8). Three variables, the initial Constant score (b = .52, 95% CI: .28-.76), the duration of disease before treatment (b = -4.4, 95% CI: from -7.2 to -1.6), and acromial morphology (b = -5.3, 95% CI: from -9.8 to -.8) were found to be independent predictors of outcome (model R2 = .68). CONCLUSION: Patients with more severe disease, a long duration of symptoms, and type II or III acromion may require more invasive therapeutic options as the first intervention. PMID- 16369369 TI - Case 8: absentminded and "walking like a drunk". PMID- 16369370 TI - Preservation of fertility in the cancer patient. PMID- 16369371 TI - Laparoscopic ovarian transposition to preserve ovarian function before pelvic radiation and chemotherapy in a young patient with rectal cancer. AB - BACKGROUND: Ovarian transposition is a surgical maneuver used to protect ovarian function before delivery of gonadocidal doses of radiation therapy. Ovarian transposition has been performed in patients whose treatment includes pelvic radiotherapy as a part of management for Hodgkin's disease and other gynecologic malignancies. CASE: Laparoscopic ovarian transposition was performed on a 28-year old female with rectal cancer. At 13-month follow-up from completion of chemoradiation treatments, normal menstrual cycles at regular monthly intervals are reported. FSH, LH, and progesterone levels obtained during the second half of menstrual cycles were within normal ranges. CONCLUSION: Ovarian transposition is an effective surgical procedure for preserving ovarian function in patients at risk of radiotherapy-induced ovarian failure. Laparoscopic techniques can be used to move the ovaries outside of the radiation portal. Consideration should be given for ovarian transposition for other abdominal and pelvic malignancies before onset of radiation treatments in patients who desire preservation of ovarian function. PMID- 16369373 TI - High prevalence of hepatitis B virus markers in Romanian adolescents with human immunodeficiency virus infection. AB - BACKGROUND: We evaluated the frequency of hepatitis coinfection in Romanian adolescents who were diagnosed with human immunodeficiency virus (HIV) infection prior to 1995. METHODS: One hundred sixty-one adolescents (13-18 years of age) with symptomatic HIV infection, but without signs of hepatic dysfunction, and 356 age-matched, HIV-uninfected controls underwent laboratory testing for markers of parenterally acquired hepatitis virus infection. RESULTS: Seventy-eight percent of HIV-infected adolescents had markers of past or present hepatitis B virus (HBV) infection, as compared with 32% of controls ( P = .0001). The prevalence of HBV replicative markers was more than 5-fold higher in HIV-infected adolescents as compared with controls: 43.4% vs 7.9% ( P = .0001), respectively, for hepatitis B surface antigen (HBsAg); and 11.2% vs 2.2% ( P = .0001), respectively, for hepatitis B e antigen (HBeAg). The prevalence of HBsAg chronic carriers and the presence of HBV replicative markers was significantly higher in patients with immunologically defined AIDS (CD4+ cell counts < 200 cells/mcL): 59.6% vs 34.6% ( P = .02) for HBsAg and 22.8% vs 5.7 %, ( P = .002) for HBV DNA. After 1 year of follow-up, the proportion of those who cleared the HBeAg was considerably lower in severely immunosuppressed coinfected patients: 4.7% vs 37.1% ( P = .003). Four additional HIV-infected adolescents became HBsAg-positive over the term of follow-up (incidence rate, 24.9/1000 person-years), despite a record of immunization against hepatitis B. CONCLUSIONS: A substantial percentage of HIV-infected and HIV-uninfected Romanian adolescents have evidence of past or present HBV infection. In HIV-infected adolescents, the degree of immunosuppression is correlated with persistence of HBV replicative markers, even in the absence of clinical or biochemical signs of liver disease. PMID- 16369374 TI - Substitutions in the reverse transcriptase and protease genes of HIV-1 subtype B in untreated individuals and patients treated with antiretroviral drugs. AB - The nucleotide transition G-->A is known as a hypermutation due to its high prevalence in HIV-1 and other pathogens. However, the contribution of the G-->A transition in the generation of drug resistance mutations is unknown . Our objective was to ascertain the rate of nucleotide substitutions in protease (PR) and reverse transcriptase (RT) in both untreated and treated HIV-1 patients. Genotypic analysis was performed on viruses from both treated and untreated patients with subtype B infections. Nucleotide genomic diversity was compared with a consensus subtype B reference virus. Then, the prevalence of resistance associated mutations in different subgroups of treated patients was evaluated in relation to the patterns of nucleotide transitions. In untreated patients (n = 50) G-->A was most prevalent, followed by A-->G, C-->T, and T-->C transitions. In treated patients (n = 51), the prevalence of A-->G was similar to that of G-->A. Among mutations that confer resistance to antiretroviral drugs, M184V was present in 76% of treated patients and K70R in 31% (A-->G transitions). Other frequent mutations in RT included T215Y (C-->A and A-->T substitutions), which was prevalent in 31% of treated patients. In PR, a L90M (T-->A substitution) was prevalent in 47% of protease inhibitor (PI)-treated patients. In conclusion, the G-->A transition was most prevalent in RT and PR among untreated patients. In contrast, A-->G was the most prevalent transition in patients treated with antiretroviral drugs. PMID- 16369375 TI - Prevalence and correlates of HIV testing: an analysis of university students in Jamaica. AB - BACKGROUND: Prevention programs often promote HIV testing as one possible strategy of combating the spread of the disease. OBJECTIVE: To examine levels of HIV testing practices among a large sample of university students and the relationship among HIV testing, sociodemographic variables, and HIV-related behaviors. METHODS: A total of 1252 students were surveyed between June 2001 and February 2002 using a 193-item questionnaire measuring a variety of HIV-related knowledge and attitudinal and behavioral items. RESULTS: Hierarchical logistic regression analyses revealed that youths, married persons, persons who had attended an HIV education forum, and those who knew someone with HIV/AIDS were more likely to report a previous HIV test. However, HIV testing was not associated with condom use or number of sex partners. CONCLUSION: The lack of significant findings between testing and risky sexual behaviors should not negate the importance of HIV testing. Being informed regarding personal HIV serostatus is one of the first steps in self-protection. Effective messages and programs need to be developed and implemented in Jamaica to promote HIV testing and help persons to adequately assess their level of risk with respect to contracting HIV. PMID- 16369376 TI - The HIV-1 Non-subtype B Workgroup: an international collaboration for the collection and analysis of HIV-1 non-subtype B data. PMID- 16369378 TI - Evidence-based management of acute myocardial infarction in the elderly--current perspectives. AB - Cardiovascular disease accounts for significant morbidity and mortality in the elderly. Despite several, large cardiovascular clinical trials, data to guide therapy in this growing population subset are relatively limited. This review focuses on treatment approaches and recommendations for the management of elderly patients with acute myocardial infarction (MI) obtained from subgroup analyses from major clinical trials.Treatment options for acute MI in the elderly have changed dramatically since the 1990s. Reperfusion therapy by primary percutaneous coronary intervention has superseded the use of thrombolytic therapy for the treatment of acute ST-elevation myocardial infarction (STEMI). Clinical trial data have demonstrated that even transferring patients to facilities that have primary angioplasty capabilities is better than thrombolytic therapy, if the anticipated transfer time is of acceptable duration. Additionally, adjunctive use of the intravenous glycoprotein (GP) receptor antagonist, abciximab, during primary angioplasty affords a reduction in the composite primary end point of death, reinfarction, and target vessel revascularization, with much of the benefit derived from the latter. Thrombolytic therapy, barring any contraindication, must be used when mechanical revascularization is not available; however, the risk for complications in the elderly is higher, especially for those 75 years and older. Studies investigating the use of thrombolytics plus GP receptor antagonists with and without percutaneous coronary intervention show questionable benefit in the elderly. PMID- 16369379 TI - Thyrotoxic atrial fibrillation. AB - Atrial fibrillation is the most common cardiac complication of hyperthyroidism and occurs in 15% of patients with hyperthyroidism. It is associated with a higher risk of thromboembolism that often involves the central nervous system. Oral anticoagulation is important in the majority of these patients to prevent thromboembolic complications. These patients require adjustment in the dose of various rate-controlling agents because of increased clearance associated with hyperthyroidism and a decrease in warfarin dosage because of increased clearance of vitamin K-dependent clotting factors. The management of thyrotoxic atrial fibrillation is summarized in this clinical review. PMID- 16369380 TI - Brain white and gray matter anatomy of MRI segmentation based on tissue evaluation. AB - Different approaches to gray and white matter measurements in magnetic resonance imaging (MRI) have been studied. For clinical use, the estimated values must be reliable and accurate when, unfortunately, many techniques fail on these criteria in an unrestricted clinical environment. A recent method for tissue clusterization in MRI analysis has the advantage of great simplicity, and it takes the account of partial volume effects. In this study, we will evaluate the intensity of MR sequences known as T1-weighted images in an axial sliced section. Intensity group clustering algorithms are proposed to achieve further diagnosis for brain MRI, which has been hardly studied. Subjective study has been suggested to evaluate the clustering group intensity in order to obtain the best diagnosis as well as better detection for the suspected cases. This technique makes use of image tissue biases of intensity value pixels to provide 2 regions of interest as techniques. Moreover, the original mathematic solution could still be used with a specific set of modern sequences. There are many advantages to generalize the solution, which give far more scope for application and greater accuracy. PMID- 16369381 TI - Serum antibodies to oral anaerobic bacteria in patients with rheumatoid arthritis. AB - BACKGROUND: This study was conducted to determine the component that causes the disease in rheumatoid arthritis (RA), which shows great resemblance to periodontitis in a pathologic context. MATERIALS AND METHODS: Within this study, the pathogen-specific IgG levels formed against Porphyromonas gingivalis FDC 381, Prevotella melaninogenica ATCC 25845, Actinobacillus actinomycetemcomitans Y4, Bacteroides forsythus ATCC 43047, and Prevotella intermedia 25611 oral bacteria were researched from the blood serum samples of 30 RA patients and 20 healthy controls with the enzyme-linked immunosorbent assay (ELISA) method. RESULTS: The IgG levels of P gingivalis, P intermedia, P melaninogenica, and B forsythus were found to be significantly higher in RA patients when compared with those of the controls. Of the other bacteria antibodies, A actinomycetemcomitans was not found at greater levels in RA serum samples in comparison with the healthy samples. CONCLUSION: The antibodies formed against P gingivalis, P intermedia, P melaninogenica, and B forsythus could be important to the etiopathogenesis of RA. PMID- 16369382 TI - Uses and limitations of bone mineral density measurements in the management of osteoporosis. AB - CONTEXT: Bone mineral density (BMD) is used to diagnose osteoporosis, and often to measure efficacy in osteoporosis treatment trials; however, there is a poor correlation between lumbar spine BMD increases and vertebral fracture risk reduction in patients receiving treatment for osteoporosis. OBJECTIVE: The purpose of this article is to review the uses and limitations of BMD measurements and the relationship between BMD and bone strength. DATA SOURCE/STUDY SELECTION: A MEDLINE literature search was conducted with the terms bone mineral density , fracture , osteoporosis , and bone strength as well as the generic names of osteoporosis therapies (alendronate, risedronate, raloxifene, teriparatide, and calcitonin). Search results were limited to English language journals and articles published within the last 20 years. Published abstracts from scientific meetings were also reviewed. CONCLUSION: BMD measurement remains the most useful diagnostic tool for identifying patients with osteoporosis. Although they are helpful in guiding decisions to initiate osteoporosis treatment, subsequent changes in BMD provide an imperfect indicator of treatment efficacy. Analyses of clinical trials show an inconsistent relationship between increased spinal BMD and a decreased risk of vertebral fracture. Increased BMD accounts for less than 25% of the overall reduction in fracture risk in most instances. Consequently, fracture risk reduction itself remains the most clinically relevant therapeutic outcome of osteoporosis therapy. PMID- 16369383 TI - A description and qualitative assessment of a 4-year intervention to improve patient counseling by improving medical student health. AB - BACKGROUND: To test whether promoting medical student health could efficiently improve patient counseling, we developed and implemented a 4-year-long curricular and extracurricular intervention to promote healthy behaviors among students in the Class of 2003 at Emory University School of Medicine, Atlanta, Georgia. METHODS: We asked students: (1) "What did you think about these [listed intervention components]"; (2) "did any of these interventions influence your personal health habits/attitudes toward your personal health"; and (3) "did any of these interventions influence your behavior or attitudes regarding current or future clinical practices, including history taking or counseling? If so, how? If not, why not?" Students evaluated the effectiveness of these formats and proposed changes in our intervention. The focus groups were transcribed and analyzed with QSR N5. RESULTS: Several major themes emerged from the focus groups: Listen to the students early, often, substantively, and noticeably; Incorporate many faculty and student leaders; Quietly integrate the curricular activities into the regular curriculum; Provide a strong, science-based, pragmatic prevention curriculum to complement the personal health promotion; Don't just use lectures to teach; Offer plentiful, nonrequired, fun extracurriculars; Don't nag; Have achievable interventions and recommendations; Provide collective data, but don't overexpose the students to it, and don't assume that collective data apply to every student, especially if it's unpleasant news; Provide personalized data where possible; and Uncouple evaluations from the intervention, and keep evaluations brief. CONCLUSIONS: Some students seemed pleased to have their medical school be attentive to their health, and believed that the project positively influenced their personal health practices and clinical practices (which was our goal). The students enjoyed many components of the intervention, especially the extracurricular activities, and recognized that they and their classmates were at risk for unhealthy behaviors that had personal and clinical implications. However, many also felt resentful and that they were nagged. Although we had anticipated these responses, and had therefore taken pains to avoid the things that the students most resented, we were insufficiently sensitive to how strong those responses would be. PMID- 16369385 TI - "Good Publication Practice for Pharmaceutical Companies": where are we now? AB - Eighteen months on from the publication of "Good Publication Practice for Pharmaceutical Companies," one member of the working group that developed these guidelines reflects on what they have achieved and what has changed since they were first developed. PMID- 16369386 TI - Seasonal variation in the prevalence of Plasmodium vivax malarial infection: an observation in Northern Thailand. PMID- 16369387 TI - Group A beta-hemolytic streptococcal glossal necrotizing myositis--case report and review. AB - We report the first case of glossal necrotizing myositis by group A beta hemolytic Streptococcus in an 8-year-old girl on chronic nonsteroidal anti inflammatory drugs, immunomodulators, and steroids for juvenile rheumatoid arthritis. Treatment included partial glossectomy and parenteral antibiotics. After a critical course, full recovery ensued. The subject of necrotizing myositis is reviewed. PMID- 16369388 TI - An accessory spleen mimics a left adrenal carcinoma. PMID- 16369389 TI - Extensive arterial and venous thrombosis in a patient with ulcerative colitis--a case report. AB - BACKGROUND: Vascular thrombosis is a rare but well-recognized extraintestinal manifestation of ulcerative colitis. Thrombosis usually involves the peripheral veins and less commonly the cerebral veins and the arterial system. CASE REPORT: A case of a 22-year-old woman is reported who presented with an exacerbation of ulcerative colitis and developed extensive arterial and venous thrombosis. Good clinical improvement was achieved after treatment with steroids, sulfasalazine, and anticoagulation with enoxaparin followed by long-term warfarin. CONCLUSION: Early recognition of thrombotic complications is essential in initiating lifesaving therapy. PMID- 16369390 TI - Iopamidol myelography-induced seizures. AB - Iopamidol, a water-soluble contrast agent, has been rarely associated with seizures. We describe a case of generalized tonic-clonic seizure after cervical myelography with iopamidol in a previously healthy young man. In patients presenting with seizures, a history of recent myelography should be considered as an etiology. Iopamidol myelography may be associated with a risk of seizures. Clinicians need to be aware of this complication and inform their patients about such risk. PMID- 16369391 TI - New Medicare rule forces cancer, heart, and lung patients into nursing homes. PMID- 16369392 TI - NSAID hysteria--Chicken Little revisited. PMID- 16369393 TI - A multisystem illness involving pancreas, bile ducts, and salivary glands. PMID- 16369394 TI - Endoscopic ultrasound-guided fine-needle aspiration in the diagnosis and staging of pancreatic adenocarcinoma. PMID- 16369396 TI - Chronic diarrhea due to metastatic breast cancer. AB - We report the case of a patient with chronic diarrhea due to infiltrative cancer to the colon from the breast. A 49-year-old woman with a history of metastatic breast cancer to the bones was admitted to hospital with 4 weeks of watery diarrhea. Computed tomography (CT) scan of the abdomen showed pancolitis. Colonoscopy showed patchy subepithelial hemorrhages, most prominently in the ascending colon and sigmoid colon, but no mass lesion. Random biopsies taken throughout the colon showed infiltrative adenocarcinoma of breast origin in multiple specimens. Infectious work-up was negative. PMID- 16369395 TI - Evaluation of perianal fistulas in patients with Crohn's disease. PMID- 16369397 TI - Applications of EUS for the molecular characterization and treatment of gastrointestinal diseases--a review of the recent medical literature. PMID- 16369398 TI - Partnering with gastroenterologists to evaluate patients with chronic constipation. AB - Constipation is a highly prevalent and bothersome disorder that negatively affects patients' social and professional lives and imposes a heavy economic burden on patients and society. Most patients with chronic constipation are evaluated and treated in the primary care setting. Primary care clinicians often underestimate how much they can accomplish in the evaluation of a patient with constipation before they make a referral. There are numerous steps that primary care clinicians can take to address these issues and maximize the benefits of the referral process, including understanding key elements of an effective diagnostic work-up, familiarizing themselves with the utility of various diagnostic tests of colonic and anorectal function, implementing strategies/instruments to optimally communicate what they are striving to achieve through the referral process (eg, via a referral form), and developing a network of long-term working relationships with local gastroenterologists. PMID- 16369400 TI - A case of polyneuropathy after gastric bypass surgery. PMID- 16369399 TI - Hepatitis B virus infection in liver transplant candidates and recipients. PMID- 16369401 TI - Gastric mass. PMID- 16369402 TI - Topics in pediatric leukemia--Fanconi's anemia: new insights. PMID- 16369404 TI - Kartagener's syndrome associated with allergic bronchopulmonary aspergillosis. PMID- 16369403 TI - A review of smoking cessation interventions. PMID- 16369405 TI - Case report: unusual mechanism of chance fracture in an adult male. PMID- 16369406 TI - Readers' responses to the webcast video editorial entitled "a healthcare system that works". PMID- 16369408 TI - Readers' responses to the webcast video editorial entitled "breaking the link: why incremental change cannot solve the issue of the uninsured". PMID- 16369410 TI - Readers' and author's responses to the webcast video editorial entitled "pandemic flu: we are not prepared". PMID- 16369411 TI - Readers' responses to the webcast video editorial entitled "should doctors, patients, and others screen for diseases?". PMID- 16369413 TI - Readers' responses to the webcast video editorial entitled "resuscitating the autopsy". PMID- 16369414 TI - Readers' responses to the webcast video editorial entitled "the American healthcare 'system' in 2005--part 7". PMID- 16369416 TI - Readers' responses to "must we fear adolescent sexuality?". PMID- 16369417 TI - Readers' responses to the webcast video editorial entitled "how to take useful drugs without hurting yourself". PMID- 16369418 TI - Readers' responses to the webcast video editorials entitled "the American healthcare 'system' in 2005"--parts 5 and 6. PMID- 16369419 TI - Readers' responses to the webcast video editorials entitled "the American healthcare 'system' in 2005"--parts 3 and 4. PMID- 16369420 TI - Reversing the trend line on childhood obesity. PMID- 16369421 TI - Help wanted: more doctors for the United States. PMID- 16369422 TI - Health information technology: improving quality and value of patient care. PMID- 16369423 TI - We can cure most breast cancers with early diagnosis and treatment. PMID- 16369424 TI - Helping families cope with sudden or unexpected death. PMID- 16369425 TI - Kids and media: learning happens. PMID- 16369426 TI - Breaking the rules: entertainment and public health. PMID- 16369427 TI - The importance of measuring quality and performance in healthcare. PMID- 16369428 TI - We must stop trying to deliver 21st-century care with a 19th-century delivery model. PMID- 16369430 TI - The real world of medical practice? PMID- 16369431 TI - A healthcare system that works. PMID- 16369432 TI - Breaking the link: why incremental change cannot solve the issue of the uninsured. PMID- 16369433 TI - Should doctors, patients, and others screen for diseases? PMID- 16369434 TI - Pandemic flu: we are not prepared. PMID- 16369435 TI - Resuscitating the autopsy. PMID- 16369436 TI - Fighting obesity--round 3. PMID- 16369437 TI - Case 9: pregnant? Who's pregnant? Memory loss in a young woman. PMID- 16369438 TI - BRCA1 and BRCA2 pathways and the risk of cancers other than breast or ovarian. AB - OBJECTIVE: Germline mutations in the tumor suppressor genes BRCA1 and BRCA2 predispose women to breast and ovarian cancer. Female carriers of BRCA1 or BRCA2 gene mutations have very high lifetime risks for breast and ovarian cancers. Genetic abnormalities occur in all cancers, so BRCA-related pathways are critical because they serve to safeguard genetic content. Although protecting genetic information is a general function, BRCA-related pathways seem largely specific to preventing breast and ovarian cancer. The objective of this study was to resolve this difference between the theoretical functions of BRCA genes and their specific clinical effects. DATA SOURCES, DATA EXTRACTION, DATA SYNTHESIS: The author collected data published in > 30 epidemiologic studies on the incidence of cancers other than breast or ovarian in mutation carriers and in large populations eligible for mutation testing. Data were extracted and used directly as published whenever possible with a minimum of statistical manipulation. CONCLUSIONS: Although mutations target breast and ovary, a broader spectrum of cancers also occur with statistically significant elevated frequencies. Risks for "all cancers except breast or ovary" are elevated, with some population subgroups differing with regard to how frequently elevated risks were found at individual sites. Additional sites at risk included stomach, pancreas, prostate, and colon. The increased risk ranged from about 20% to 60%, with the greatest increases in risk in stomach and pancreas. The collected data show BRCA-pathway functions are probably required at multiple sites, not just in breast or ovary. Known interactions and relationships among BRCA-related pathways strongly support the idea that their inactivation provides growth or survival advantages for a variety of cancers. The data suggest applying an increased level of clinical alertness to those with defects in BRCA-related pathways. Identifying molecules that confer growth or survival advantages to BRCA-related cancers may provide broadly useful targets for chemotherapy or chemoprevention. PMID- 16369439 TI - How do clinicians manage vaginal complaints? An Internet survey. AB - INTRODUCTION: Although vaginal symptoms are among the most common presentations in primary care, little is known about how clinicians manage them. QUESTION: We undertook an Internet survey to determine how clinicians decided what was normal, how they evaluated vaginal complaints, and what happened when a diagnosis could not be reached. METHODS: A 27-question anonymous survey was conducted using the Medscape Internet publication platform and Medscape General Medicine , an online, peer-reviewed, open-access general medical journal. The study was targeted at physicians and nurse practitioners. RESULTS: There were 556 nurse practitioner and 608 physician respondents. Clinicians generally agreed that a midcycle increase in discharge and discharge seen at the introitus were normal; itching/irritation, odor, and a green discharge were not. There was less agreement concerning the value of a yellow discharge and "self-diagnosis" of abnormality. Eighty-two percent of MDs and 77% of NPs reported examining patients prior to treatment always or often. Only about 30% performed pH testing always or often. Ninety percent of NPs and 92% of MDs reported not making a diagnosis rarely or sometimes. When offered 4 different management strategies for patients in whom no diagnosis was made (watch and wait, symptomatic therapy, empiric antibiotics, referral), there was no clear preference. There was a weak correlation between performance of a wet mount and reports of less non-diagnosis. There was no correlation between pH and whiff testing and reports of less non diagnosis. CONCLUSIONS: Clinicians do not always agree about what constitutes a vaginal symptom. Although most providers usually examine patients with vaginal complaints, pH and whiff testing are performed less frequently. Our data suggest the latter 2 maneuvers are not diagnostically helpful. There is no consensus regarding what to do when no diagnosis is made. Our findings suggest that the traditional approach to the evaluation of vaginal symptoms may have limited relevance for clinicians. PMID- 16369440 TI - Politics, power, and procreation. PMID- 16369441 TI - Recurrent preterm abruption--case report. PMID- 16369442 TI - Cervical endometriosis, a case presenting with intractable spotting. AB - Cervix uteri is regarded as an infrequent localization for endometriosis. With widespread use of invasive cervical procedures, however, an increased incidence can be expected. Limited awareness of the clinical appearance of the disease may account for its apparent rarity. This presentation aims to refocus attention to the disease by reviewing the case of a woman who presented to us with minimal metrorrhagia, which is a rare symptom of cervical endometriosis. PMID- 16369443 TI - Where is HIPAA taking physician practices? PMID- 16369444 TI - Clinical data capture: OMR and OCR and your flatbed scanner. PMID- 16369445 TI - A 28-year-old woman presented to Kijabe Mission Hospital with severe dyspnea. PMID- 16369447 TI - Migration and risk factors for HIV acquisition in pregnant women in Baja California, Mexico. PMID- 16369448 TI - Triple-nucleoside analog antiretroviral therapy: is there still a role in clinical practice? A review. AB - The development and widespread clinical use of coformulated abacavir/lamivudine/zidovudine (ABC/3TC/ZDV) as Trizivir represented an important advance in the management of HIV-infected patients, especially those with adherence challenges. With a low pill burden, no food restrictions, limited drug drug interactions, and a favorable resistance profile, ABC/3TC/ZDV remains an alternative option in the US Department of Health and Human Services Consensus Panel Guidelines as initial treatment in antiretroviral-naive patients. Recent data have shown ABC/3TC/ZDV to be less efficacious in suppressing and/or maintaining suppression of virologic replication compared with efavirenz containing antiretroviral therapy. Although triple-nucleoside/nucleotide reverse transcriptase inhibitor (t-NRTI) combinations that do not contain a thymidine analog (ZDV or stavudine) have recently shown high virologic failure rates in clinical trials and clinical practice, t-NRTI regimens containing a thymidine analog have consistently been shown to be efficacious. PMID- 16369449 TI - Cytokine profiles in human immunodeficiency virus-infected children treated with highly active antiretroviral therapy. AB - CONTEXT: There have been few longitudinal studies of cytokine production in neonatally acquired HIV-1 infection and none in Asian or Chinese children. OBJECTIVE: To determine whether monitoring cytokine production could contribute to the better management of pediatric patients with HIV-1 infection. SETTING: Clinical Immunology Laboratory and Pediatrics Department, University Hospital, Hong Kong. PATIENTS: Ten Asian and 2 Eurasian children infected with HIV-1 by mother-to-child transmission were followed for up to 5 years while on treatment with highly active antiretroviral therapy (HAART). MAIN OUTCOME MEASURES: Numbers of unstimulated and mitogen-activated cytokine-secreting cells (IFN-gamma, interleukin [IL]-2, IL-4, IL-6, IL-10, IL-12, and TNF-alpha) were measured by ELISPOT assay at frequent intervals, and correlations were sought with CD4+ and CD8+ cell counts and viral loads. RESULTS: Mitogen-stimulated IL-2-secreting cells were directly associated with recovery of CD4+ cells. Correlations with viral load were found for Con A-induced IFN-gamma, Con A-induced IL-4, and unstimulated IL-10, suggesting that these cytokines were either suppressed by high virus levels or that higher cytokine levels suppressed virus. IFN-gamma, IL 2-, IL-4-, and IL-12-secreting cells induced by PHA, Con A, and/or SAC tended to increase for the first 3-4 years of treatment but declined thereafter. CONCLUSIONS: Alterations in cytokine profiles were not associated with adverse clinical events and there was little evidence to indicate that monitoring cytokine enzyme-linked immunospots (ELISPOTs) could contribute to pediatric patient management. PMID- 16369450 TI - B-cell depletion for rheumatic diseases: where are we? AB - Immune system dysfunction is common to rheumatic disorders, with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) being classic examples. Altered development and function of B cells may play a prominent role. B-cell abnormalities also occur in other rheumatic diseases, eg, Sjogren's syndrome, Behcet's disease, antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, and dermatomyositis. Hence, B-cell depletion has been investigated as a therapeutic option. Clinical trials in RA and SLE have shown that rituximab, an anti-CD20 monoclonal antibody, can profoundly reduce disease activity and is generally well tolerated. Reports of rituximab treatment for ANCA-associated vasculitis and dermatomyositis are also promising. These encouraging results validate the strategy of B-cell depletion in various rheumatic diseases. B-cell depletion with rituximab is under study in larger clinical trials for the purposes of regulatory approval to define more closely its place in RA and SLE treatment paradigms, and smaller clinical trials are ongoing or planned in associated inflammatory diseases. PMID- 16369451 TI - The prevention and treatment of osteoporosis: a review. AB - Osteoporosis is a disorder characterized by reduced bone strength, diminished bone density, and altered macrogeometry and microscopic architecture. Adult bone mass is the integral measurement of the bone mass level achieved at the peak minus the rate and duration of subsequent bone loss. There is clearly a genetic predisposition to attained peak bone mass, which occurs by a person's mid-20s. Bone loss with age and menopause are universal, but rates vary among individuals. Both peak bone mass and subsequent bone loss can be modified by environmental factors, such as nutrition, physical activity, and concomitant diseases and medications. Osteoporosis prevention requires adequate calcium and vitamin D intake, regular physical activity, and avoiding smoking and excessive alcohol ingestion. Risk of fracture determines whether medication is also warranted. A previous vertebral or hip fracture is the most important predictor of fracture risk. Bone density is the best predictor of fracture risk for those without prior adult fractures. Age, weight, certain medications, and family history also help establish a person's risk for osteoporotic fractures. All women should have a bone density test by the age of 65 or younger (at the time of menopause) if risk factors are present. Guidelines for men are currently in development. Medications include both antiresorptive and anabolic types. Antiresorptive medications- estrogens, selective estrogen receptor modulators (raloxifene), bisphosphonates (alendronate, risedronate, and ibandronate) and calcitonins--work by reducing rates of bone remodeling. Teriparatide (parathyroid hormone) is the only anabolic agent currently approved for osteoporosis in the United States. It stimulates new bone formation, repairing architectural defects and improving bone density. All persons who have had osteoporotic vertebral or hip fractures and those with a bone mineral density diagnostic of osteoporosis should receive treatment. In those with a bone mineral density above the osteoporosis range, treatment may be indicated depending on the number and severity of other risk factors. PMID- 16369452 TI - Choice of antihypertensive drug in the diabetic patient. AB - The hypertensive patient with type 2 diabetes is especially at risk of adverse cardiovascular events. The United Kingdom Prospective Diabetes Study (UKPDS) and Hypertension Optimal Treatment (HOT) studies suggested that treatment to a lower target blood pressure resulted in better prevention of clinical disease in these patients. Most trials comparing antihypertensive drugs have shown only minimal differences between the various agents. The evidence from the trials suggests that diuretics, beta-blockers, calcium channel blockers (CCBs), angiotensin converting enzyme (ACE) inhibitors, and the angiotensin-receptor antagonists (ARBs) will all successfully reduce adverse clinical events. The largest of the comparative hypertensive drug trials, the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT), demonstrated that a diuretic has a better hypotensive effect, and was more successful in preventing many aspects of cardiovascular disease compared with CCBs and ACE inhibitors. The importance of good blood pressure control and the general equivalence of antihypertensive drugs were again shown in the Valsartan Antihypertensive Long term Use Evaluation (VALUE) trial, which compared an ARB with a CCB. Choice of antihypertensive agent should be individualized and guided by the presence of concomitant clinical disease and the need to protect any specific target organ system in the diabetic hypertensive. Diuretics, being potent hypotensive drugs with clearly demonstrated clinical benefit, should form part of the antihypertensive regimen of most diabetic hypertensives. ACE inhibitors and ARBs are especially useful in preventing nephropathy. Most patients will require a combination of antihypertensive drugs to achieve tight blood pressure control of under 130/80 mm Hg in the diabetic hypertensive. The clinician should concentrate on seeking this lower target blood pressure rather than be excessively concerned about which is the best antihypertensive agent. PMID- 16369453 TI - Evidence-based management of coronary artery disease in the elderly--current perspectives. AB - Cardiovascular disease accounts for significant morbidity and mortality in the elderly. The clinical trial data available to guide therapy in this growing population subset are relatively limited. This review will focus on treatment approaches and recommendations obtained from subgroup analyses of elderly patients from major clinical trials for the management of chronic stable angina, acute coronary syndromes (unstable angina and non-ST-segment elevation myocardial infarction), and coronary revascularization. Recent advances in the treatment of stable angina have shown that use of angiotensin-converting enzyme inhibitors and lipid-lowering therapy as adjunctive measures show benefit in the elderly by reducing the occurrence of death, nonfatal myocardial infarction, and unstable angina. However, if patients experience disabling or unstable anginal symptoms despite effective medical therapy, coronary revascularization must be considered. Several clinical trials have shown a significant reduction in major adverse cardiac events when using intravenous glycoprotein receptor antagonists periprocedurally during percutaneous revascularization approaches in elderly patients with unstable angina or non-ST-segment elevation myocardial infarction, especially when these measures are performed as soon as possible. However, the success of myocardial revascularization by a percutaneous or surgical approach is highly dependent on the patient's associated comorbidities, especially in patients over age 80 years. PMID- 16369454 TI - A Critique of the USMLE clinical skills examination. PMID- 16369455 TI - Editorial: The learning curve. PMID- 16369456 TI - International electives: maximizing the opportunity to learn and contribute. PMID- 16369457 TI - Stimulation by bradykinin of HCO3- secretion in rat gastroduodenal mucosa. AB - BACKGROUND: We examined the effect of bradykinin on gastroduodenal HCO(3)(-) secretion in rats and investigated the mechanisms involved in this action. MATERIAL/METHODS: Under urethane anesthesia, a chambered stomach or a proximal duodenal loop was perfused with saline, and the secretion of HCO(3)(-) was measured at pH 7.0 using a pH-stat method and by adding 2 mM HCl. RESULTS: Intravenous administration of bradykinin increased both gastric and duodenal HCO(3)(-) secretion in a dose-dependent manner. These effects were totally blocked by FR172357, the bradykinin B2 receptor antagonist, and significantly attenuated by indomethacin or L-NAME, although the degree of inhibition was much greater in the stomach than the duodenum. Likewise, the response to bradykinin in the stomach totally disappeared on the chemical ablation of capsaicin-sensitive afferent neurons, whereas this action in the duodenum was inhibited only partially by sensory deafferentation. Capsazepine, the antagonist of transient receptor potential vanilloid type 1 (TRPV1), did not significantly affect the HCO(3)(-) response to bradykinin in these tissues. CONCLUSIONS: Bradykinin increases both gastric and duodenal HCO(3)(-) secretion through the activation of B2 receptors, and this action is mediated locally by endogenous prostaglandins (PGs) and nitric oxide (NO) as well as capsaicin-sensitive afferent neurons. It is assumed that bradykinin causes the release of PGs and NO as well as activation of afferent neurons via B2 receptors but not through the interaction with TRPV1, and these factors are all involved in the gastroduodenal responses, although the mode of interaction between these factors may be different in the stomach and duodenum. PMID- 16369458 TI - Macrophage may responses to androgen via its receptor. AB - BACKGROUND: Sex hormones have profound effects on immune responses and may influence the disease which caused by intracellular parasite(Leishmania) and bacterial (tuberculosis)and also autoimmune disease such as rheumatoid arthritis (RA). It has also been demonstrated that 5alpha-Dihydrotestosterone (5alpha-DHT) modulate nitric oxide and cytokine release by macrophages. These effects seem to be exerted by specific receptors for androgen in macrophages. MATERIAL/METHODS: Protein secretion: The effect of 5alpha-DHT on protein secretion by peritoneal macrophages of NZB?BALBc mice was investigated using radiolabelled protein secretion following SDS-PAGE and Fluorography. Binding assay: Androgen binding was also investigated using an autoradiography method. Peritoneal macrophages were treated with [3H]- 5alpha-DHT and incubated for 2 h before smearing on to microscope slides. Slides were air dried, dipped in Kodak NTB photographic emulsion, sealed in light proof boxes and left at 4 degrees C for 6 weeks. RESULTS: The results showed that protein secretion by macrophages changed under 5alpha-DHT treatment. Analysis of the data according to quantitation of [(3)H] 5alphaDHT binding receptors in fixed-slide mounted cells, identified a high specific androgen binding at physiological concentration. The receptors had a relatively high affinity for the 5alpha-DHT, So that binding affinity was not inhibited in the presence of 100-fold excess of non labelled 17-beta Estradiol. CONCLUSIONS: These results suggest that the immunosuppressive action exerted by androgen is at least partially achieved through a direct influence on macrophages. PMID- 16369459 TI - In vivo assessment of epichlorohydrin effects: the chorioallantoic membrane model. AB - BACKGROUND: Previous studies on the effects of the epichlorohydrin (ECH) epoxide demonstrated this compound's toxicity and mutagenicity and suggested a carcinogenic activity also in humans. To gain a better understanding of ECH effects in vivo, the substance was tested on developing tissues utilizing the chick embryo chorioallantoic membrane (CAM) assay. MATERIAL/METHODS: Gelatin sponges adsorbed with ECH were implanted onto nine-day CAMs. After five days the membranes were fixed, cut in serial sections, and stained with toluidine blue. Sections of the ECH-treated CAMs were also submitted to immunocytochemistry for the basal lamina glycoprotein laminin and the gap junction protein connexin 43 (Cx43). Control CAMs were treated with saline solution and submitted to identical procedures. RESULTS: ECH-treated CAMs displayed proliferation of both the epithelial layers and the mesenchyme cells and vessels. The laminin immunolabeling was interrupted beneath the ectoderm thickenings, which penetrated the mesenchyme. The endoderm showed papilloma-like formations and its laminin positive basal membrane protruded toward the mesenchyme, together with clusters of endodermal cells. The mesenchyme showed increased numbers of cells and microvessels. These reactions were restricted to regions corresponding to the implant. Cx43 expression was strongly decreased in the ECH-treated CAMs compared with the controls, where the connexin punctate pattern regularly decorated the epithelial cell contours. CONCLUSIONS: The study confirms that ECH elicits tissue proliferation at the contact site and corroborates the suggestion of an ECH carcinogenic effect due to hallmarks of tumoral growth, such as angiogenesis, basal membrane alterations, and loss of intercellular communication via gap junctions. PMID- 16369460 TI - Acute effects of exogenous nitric oxide on glucose uptake in skeletal muscle of normoglycaemic and diabetic rats. AB - BACKGROUND: The role of nitric oxide (NO) in the modulation of basal and insulin stimulated glucose uptake remains controversial. The aim of this study was to investigate the role of NO released from its donors, S-nitrosoglutathione (GSNO) and S-nitroso-N-acetylpenicillamine (SNAP), on glucose uptake in skeletal muscle of normoglyceamic and type 2 diabetic rats. MATERIAL/METHODS: Skeletal muscle strips of type 2 diabetic and normoglycaemic Sprague-Dawley rats were incubated with or without various concentrations (200 microM, 500 microM, 1000 microM, 10 mM & 20 mM) of SNAP or GSNO in the presence or absence of insulin (100 nM or 10 microM). The associated radioactivity was determined by liquid scintillation counting in a Beckman LS6000 scintillation counter programmed for dual-channel counting. RESULTS: SNAP and GSNO at 1000 microM significantly elevated basal and insulin-stimulated 2-deoxyglucose uptake (P<0.05) in normoglycaemic and diabetic rats. Millimolar concentrations (10 & 20 mM) of GSNO and SNAP significantly decreased basal and insulin-stimulated glucose uptake in skeletal muscle strips of normoglycaemic and type 2 diabetic rats in a concentration-dependent manner (P<0.05). The inhibition of insulin-stimulated glucose uptake was greater in the diabetic rats using both NO donors compared with normoglycaemic rats (P<0.05). CONCLUSIONS: The stimulatory effect of micromolar concentrations of GSNO and SNAP enhances basal and insulin-stimulated glucose uptake in normoglycaemic and type 2 diabetic rats, however higher concentrations elicited an inhibitory effect in normoglycaemic and diabetic rats. This highlights the NO-glucose uptake mechanism as a possible potential therapeutic target in the treatment of type 2 diabetes. PMID- 16369461 TI - The influence of gastric pentadecapeptide BPC 157 on acute and chronic ethanol administration in mice. The effect of N(G)-nitro-L-arginine methyl ester and L arginine. AB - BACKGROUND: Alcohol disturbances, NO stimulation (by the NO-precursor L arginine), and/or NO-synthesis blockade (by N(G)-nitro-L-arginine methyl ester, i.e. L-NAME) were challenged with stable gastric pentadecapeptide BPC 157, which inhibits both acute alcohol intoxication and alcohol withdrawal symptoms. MATERIAL/METHODS: Mice received intraperitoneally (i.p.) BPC 157 (10 microg/kg), L-NAME (10 mg/kg), and L-arginine (400 mg/kg), alone or in combination, 5 minutes before or after acute ethanol (4 g/kg i.p.) intoxication or after 0, 3, or 7 hours of withdrawal after drinking 20% alcohol for 13 days. RESULTS: BPC 157 rapidly opposes the strongest disturbance presentations in acute intoxication (sustained ethanol anesthesia, complete loss of righting reflex, no reaction to external stimuli, hypothermia, 25% mortality) and withdrawal (prominent seizures). NO-agents: Aggravation of acute alcohol intoxication and opposition to withdrawal are common, but the later intervals affected by L-arginine and the action throughout the experiment by L-NAME are distinctive. Given together, L arginine and L-NAME counteract each other, while either the "L-NAME presentation" (acute intoxication) or the "L-arginine presentation" (withdrawal) predominates. BPC157+NO-agent: In acute intoxication (L-NAME predominating in NO-system functioning to aggravate intoxication), both BPC157+L-NAME and BPC157+L-arginine follow the presentation of L-NAME, but without worsened mortality. In withdrawal (L-arginine predominating in NO-system functioning to oppose disturbance symptoms), BPC157+L-NAME follows the presentation of L-NAME, while BPC 157+L arginine imitates that of L-arginine. CONCLUSIONS: The relationships among pentadecapeptide BPC 157, the NO-system, acute alcohol intoxication, and opposed withdrawal may be important, presenting pentadecapeptide BPC 157 as a suitable alcohol antagonist. PMID- 16369462 TI - Differential neuronal expression of manganese superoxide dismutase in Alzheimer's disease. AB - BACKGROUND: Manganese-dependent superoxide dismutase (MnSOD) is a major defense mechanism against potential cellular damage by reactive oxygen species (ROS). We have reported increased lipid peroxidation and decreased Cu/Zn SOD enzymatic activity in the temporal cortex of Alzheimer's diseased (AD) brains. MATERIAL/METHODS: We now report the expression of MnSOD in the hippocampus of AD (n=8) and non-AD patients (n=7) via immunohistochemical methods, in both neuronal and non neuronal cells. We tested the hypothesis that there is differential expression of MnSOD in the CA1, CA2/3, and CA4 region of the hippocampus which may account for the neuronal loss seen in Alzheimer's disease. For neuronal cells, profile counts were made and expressed as positive neuronal profiles (MnSOD-IR) per mm(2) within hippocampal regions CA1, CA2/3, and CA4. RESULTS: The AD MnSOD-IR counts were over 9-fold higher in the CA2/3 region (p<0.001) and over 11-fold higher in the CA4 (p<0.001) region when compared to non-AD samples. The CA1 region in the AD samples showed the smallest increase (3-fold; p<0.05) when compared to non-AD patients. CONCLUSIONS: Since the CA1 region in AD is the most severely affected by the disease, our results suggest that normal compensatory mechanisms may be insufficient to protect this region from free radical oxidative damage. PMID- 16369463 TI - Association between oxygen consumption and nitric oxide production during the relaxation response. AB - BACKGROUND: Mind/body practices that elicit the relaxation response (RR) are currently practiced by over 30% of American adults. RR elicitation reduces volumetric oxygen consumption (VO(2)) from rest and counteracts the effects of stress, although the mechanisms mediating the RR remain unknown. This study was designed to investigate whether RR elicitation is mediated by nitric oxide (NO). We developed a method to quantify depth of RR using change in VO(2) (slope) during RR elicitation. We evaluated whether depth of RR elicitation was correlated with changes in NO, as measured by percentage changes in fractional exhaled nitric oxide (F(E)NO). MATERIAL/METHODS: We conducted a randomized, controlled trial, in which 46 subjects were randomized to either 8-weeks of RR training using audiotapes (n=34) or 8-weeks of exposure to a control condition- receiving health-education by audiotapes (n=12). Prior to randomization, VO(2) and F(E)NO were measured while subjects listened to a control audiotape. Eight weeks later, VO(2) and F(E)NO were measured while the RR group listened to a RR eliciting audiotape and the control group listened to a control audiotape. RESULTS: Prior to receiving any training, there was no association between VO(2) slope and F(E)NO. After training, there was an inverse correlation between VO(2) slope and F(E)NO in the RR group (r = -0.41, P=0.037, n=26), but not in the control group (r=0.12, P=0.78, n=8). CONCLUSIONS: Depth of RR elicitation was associated with increased concentrations of F(E)NO after RR training. The RR may be mediated by NO helping to explain its clinical effects in stress-related disorders. PMID- 16369464 TI - Relationship of serum immunoglobulin and IgG subclass levels to race, ethnicity and behavioral characteristics in HIV infection. AB - BACKGROUND: To determine whether demographic and behavioral factors affect immunoglobulin regulation in HIV infection, we studied injection drug users, women, and minority ethnic and racial groups with and without HIV infection. MATERIAL/METHODS: A prospective cross-sectional study of ambulatory persons with or at risk for HIV infection was conducted. We enrolled 48 injection drug users (IDUs) and 43 non-IDUs seropositive for HIV and 22 seronegative at-risk individuals in the Bronx, New York City. Sixteen HIV-seronegative, non-IDUs controls were also studied. Total serum immunoglobulin levels, IgG subclasses and lymphocyte phenotypes were measured. RESULTS: Serum IgG, IgA, IgG(1) and IgG(3) were increased in all stages of HIV infection controlling for injection drug use, gender, race and age (p0.05). Serum IgM levels were significantly decreased in HIV seropositives compared to HIV seronegatives (p<0.02). Two patterns of serum immunoglobulin level elevation were found in HIV infection: 1) IgG, IgG(1) and IgG(3) levels were elevated in early and advanced HIV infection; 2) IgA, IgG(2) and IgG(4) levels were elevated only in advanced HIV infection. IgG levels were increased in Blacks compared to Caucasians with HIV infection (p=0.01). CONCLUSIONS: Serum IgG, IgG(1) and IgG(3) levels are increased in early HIV infection, while serum IgA, IgG(2), and IgG(3) levels are increased only in advanced HIV infection. In contrast, serum IgM levels are decreased in HIV infection. HIV-seropositive Blacks have higher serum IgG levels than HIV seropositive Caucasians. Further studies are necessary to determine the mechanism(s) underlying the different patterns of immunoglobulin elevation in HIV infection. PMID- 16369465 TI - Decreased levels of plasma adiponectin in prediabetes, Type 2 diabetes and coronary artery disease. AB - BACKGROUND: Adiponectin, an adipocyte-derived hormone is implicated in insulin resistance and atherosclerosis. The aim of our study is to compare the levels of adiponectin in subjects with prediabetes and diabetes with subjects with normal glucose. MATERIAL/METHODS: In a cross sectional study, we measured adiponectin and insulin in addition to fasting lipid panel and glucose in 28 subjects with prediabetes, 28 subjects with normal fasting glucose and 33 subjects with type 2 diabetes. RESULTS: Adiponectin levels negatively correlate with insulin resistance index (r = -32.7). There is a significant difference in the levels of adiponectin between subjects with prediabetes and normal fasting glucose. There is a significant difference in adiponectin levels in subjects with prediabetes with Coronary artery disease (CAD) compared to those without CAD (10.24+/-1.02 vs. 7.02+/-0.92 with p<0.01). There is a significant difference in adiponectin levels in subjects with CAD compared to those without coronary artery disease (9.94+/-1.02 vs. 5.02+/-0.82 with p<0.01). CONCLUSIONS: We conclude that decreased plasma adiponectin and insulin resistance coexist in subjects with prediabetes, diabetes and atherosclerosis. Adiponectin concentration is an independent correlate of insulin resistance and atherosclerosis. PMID- 16369466 TI - The impact of rosiglitazone on cardiovascular responses and endurance during isometric exercise in patients with Type 2 diabetes. AB - BACKGROUND: Vascular endothelial function is compromised in people with diabetes reducing tissue blood flow. The present investigation was a logical extension of a previous work, examining isometric exercise performance and recovery in people with diabetes and the impact of the insulin sensitizer rosiglitazone, a drug shown to improve endothelial function. MATERIAL/METHODS: Blood pressure, heart rate and limb blood flow were examined in 12 control subjects and 8 subjects with type 2 diabetes during a series of 2 fatiguing isometric contractions at a tension of 40% of the maximum strength of the handgrip muscles; 5 minutes of rest were allowed between the contractions. RESULTS: Endurance of the first and the second contraction was significantly less in subjects with diabetes (p<0.01). HR increased significantly (p<0.01) by over three fold during exercise in control subjects compared to subjects with diabetes. There was no statistical difference in the increase in either systolic or diastolic BP during isometric exercise comparing controls to subjects with Type 2 diabetes (p>0.05), however, resting and peak blood pressures were greater in subjects with type 2 diabetes (p<0.01). Forearm blood flow, which was significantly lower at rest (p<0.01) in subjects with diabetes, only increased by about half as much during the exercise in subjects compared to control subjects. These differences were partially reversed after 3 months administration of rosiglitazone. CONCLUSIONS: Microcirculatory damage in diabetes impairs exercise performance and recovery and can be partially reversed after rosiglitazone administration. PMID- 16369467 TI - Study of insulin-like growth factor-1 (IGF-1) and platelet-derived endothelial cell growth factor (PDEGF) expression in children with infantile hypertrophic pyloric stenosis. AB - BACKGROUND: The pathogenesis of infantile hypertrophic pyloric stenosis (IHPS) is not fully understood. Hypertrophy of the pyloric muscle is probably regulated by growth factors. Recent studies reported an increase in the local synthesis of insulin-like growth factor-1 (IGF-1). There are no reports concerning platelet derived endothelial cell growth factor (PDEGF) playing an important role in the pathological angiogenesis. The aim of this study was to analyze the expressions of IGF-1 and PDEGF by immunohistochemistry (IHC) in the muscularis propria of the pyloric muscle in children with IHPS. MATERIAL/METHODS: Twenty-two muscle biopsies were obtained at the time of pyloromyotomy. The control group consisted of seven children. Specimens were evaluated by routine histopathological methods and by immunohistochemistry using monoclonal mouse anti-PDEGF or -IGF-1 antibodies. Cells showing positive reaction were counted in five random 200x high power fields. Values were expressed as the mean +/-SD of the real expression area of the analyzed marker to the total analyzed area. RESULTS: In children with IHPS the average area of PDEGF expression was 62+/-52.5, whereas in the control group it was 15+/-12.1. The average area of IGF-1 expression was 1037+/-491.9) in study group and 259+/-221.44 in the controls. Statistically significant differences were found. CONCLUSIONS: These results show a local increase in the expressions of IGF-1 and PDEGF in the muscularis propria of the pyloric muscle in children with IHPS, which may have implications to the pathogenesis of the disease. PMID- 16369468 TI - Surgical technique and postoperative complications in congenital cataract surgery. AB - BACKGROUND: We evaluated cases that underwent congenital cataract surgery in terms of the surgical technique, post-operative complications, and visual prognosis. MATERIAL/METHODS: We retrospectively evaluated 132 eyes in 76 patients with congenital cataracts who underwent cataract surgery between January 1995 and December 2000. The patient age at surgery, surgical technique, post-operative complications, and final visual prognosis were evaluated. RESULTS: Thirty eyes underwent aspiration, posterior capsulotomy, and intraocular lens (IOL) implantation; 22 eyes underwent aspiration and posterior capsulotomy; 39 eyes underwent aspiration, posterior capsulotomy, and anterior vitrectomy; and 41 eyes underwent aspiration, posterior capsulotomy, anterior vitrectomy, and IOL implantation. The most frequent complications were secondary cataract (24.2%), posterior synechiae (9.1%), and glaucoma (3%). Groups were compared according to surgical technique and incidence of complications. Complications occurred most frequently with aspiration and posterior capsulotomy and occurred less frequently with aspiration, posterior capsulotomy, and anterior vitrectomy; these differences were statistically significant (p<0.05). The difference in the final visual prognosis was statistically significant in the aspiration, posterior capsulotomy, anterior vitrectomy, and IOL implantation groups as compared with the other groups (p<0.05). The mean follow up times was 15.5+/-1.78 months. CONCLUSIONS: The most frequent complication of congenital cataract surgery is secondary cataract formation. Today, posterior capsulotomy with anterior vitrectomy is the preferred method for decreasing the incidence of this complication. PMID- 16369469 TI - Prevalence and risk factors for HEV infection in pregnant women. AB - BACKGROUND: Hepatitis E is an infectious viral disease with clinical and morphological features of acute hepatitis, clinically similar to other forms of acute viral hepatitis except in pregnant women, in whom the illness is particularly severe and has a high mortality rate. The present study was conducted in western Turkey to investigate the prevalence and risk factors for HEV infection in pregnant women. MATERIAL/METHODS: The data for the study were acquired from health centers in urban and rural areas of Aydin province, Turkey. The study design was cross-sectional. Multistage sampling was used to select the study group. Samples were tested for anti-HEV IgG by commercial ELISA test. RESULTS: A total of 386 pregnant women were included in the study. Antibodies against HEV were detected in 27 of the 386 pregnant women (7.0%). The prevalence of HEV seropositivity was significantly lower (2.5%) in women with a higher education level when compared to women with a lower education level (9.7%) (p=0.023). No significant differences were identified between seropositive and seronegative women in terms of age, source of water supply and place of residence. CONCLUSIONS: According to our results, education seems to be the only factor affecting the prevalence of HEV infection in pregnant women. The prevalence rate we found was similar to the results obtained in previous community-based studies conducted in western Turkey. PMID- 16369470 TI - Does maternal height affect triplets' birth weight? AB - BACKGROUND: In cases of triplet gestation where patients are reluctant to undergo multifetal pregnancy reduction, it would be helpful to identify predictive factors regarding poor or better outcomes. One such possible factor may be maternal height, which is possibly predictive of gestational age and neonatal birth weight. MATERIAL/METHODS: To examine such a possible association, we have retrospectively evaluated 102 triplet gestations. Maternal height and BMI were compared and correlated to neonatal weight, week of delivery, NICU hospitalization duration, and other parameters of pregnancy outcome. RESULTS: Mothers taller than 165 cm gave birth to significantly heavier neonates than shorter parturients delivered of triplets. Individual and mean total triplet neonatal weights were positively correlated to maternal height. There was no significant correlation between preconceptional maternal BMI and triplet neonatal weight and week of delivery, NICU hospitalization or any other parameter. CONCLUSIONS: The taller patient (>165 cm) may be at a significantly lower risk of very low birth weight neonates and very premature delivery as compared to the shorter patient (< 165 cm). Therefore, the factor of maternal height may be taken into consideration in multiple gestation pregnancy consultations. Smaller mothers should never receive more than two embryos in IVF programs to reduce the risk of triplets almost completely. PMID- 16369471 TI - Membranous interventricular septal aneurysm and associated atrioventricular block causing symptoms in a 65-year-old man. AB - BACKGROUND: Membranous interventricular septal aneurysm has been reported to accompany atrioventricular (AV) conduction defects. If it is accompanied by AV block, patients generally become symptomatic before the age of 40. CASE REPORT: A 65-year-old male presented with presyncope and extreme fatigue for one week. Resting ECG showed a 2: 1 AV block. Transthoracic echocardiography revealed an interventricular septal aneurysm without any shunt. After making the diagnosis of type 2 second-degree AV block, a DDDR pacemaker was implanted. CONCLUSIONS: We presented a 65-year-old male with new-onset presyncope. A diagnosis of septal aneurysm and associated AV block was made and a pacemaker was implanted. PMID- 16369472 TI - Budd-Chiari syndrome: current options in interventional radiology treatment exemplified by three selected cases. AB - BACKGROUND: Budd-Chiari syndrome (BCS) is rare clinical state characterized by stenosis or complete obstruction of hepatic veins. Currently, interventional radiology techniques are more frequently used as a single method of treatment or as a bridge to liver transplantation. CASE REPORT: This study presents current interventional radiology techniques used in BCS treatment. Depending on the etiology of BCS, two main techniques are used: the transjugular intrahepatic portocaval shunt (TIPS) or percutaneous angioplasty (PTA) of the stenosed hepatic veins. Our first case was treated by PTA of the stenosed ostium of the hepatic vein. In the second, BCS was complicated by portal vein thrombosis and a TIPS was placed along with portal vein fibrynolysis. In the third case the TIPS was used as a single interventional radiological treatment. CONCLUSIONS: TIPS placement or angioplasty of hepatic vein ostium stenosis allow the successful treatment of BCS or an extension of the period of waiting for a liver transplantation. PMID- 16369473 TI - Could new biotechnologies revive the old moral principles? PMID- 16369474 TI - Prevention of motion sickness with rizatriptan: a double-blind, placebo controlled pilot study. AB - BACKGROUND: Serotonergic triptan medications effectively reverse pain and associated symptoms of migraine. Vestibular symptoms, including dizziness and vertigo, occur in about one-third of migraineurs. The most frequent and consistent balance symptom in migraineurs is motion sickness, which is experienced by about half of migraineurs and may be related to serotonergic influences. This double-blind, placebo-controlled, crossover pilot study was designed to test the hypothesis that pre-treatment with the serotonin agonist rizatriptan would prevent motion sickness provocation in headache-free migraineurs. MATERIAL/METHODS: Ten healthy adult migraineurs (5 migrainous vertigo and 5 migraine without associated vestibular symptoms) with a history of motion sickness were tested in three sessions: a baseline vestibular battery and two motion sickness provocation sessions two hours following randomly ordered blinded pre-treatment with either oral rizatriptan 10 mg or placebo. Motion sickness was assessed using two standardized questionnaires. RESULTS: Motion sickness scores were lower following pre-treatment with rizatriptan compared with placebo in subjects with migrainous vertigo. Rizatriptan did not affect motion sickness in migraineurs without vertigo. Otolith-ocular reflex sensitivity was reduced following pretreatment with rizatriptan in both groups. CONCLUSIONS: This is the first study in humans testing a triptan serotonin agonist as a preventive tool for motion sickness. Rizatriptan prevented the development of motion sickness and severe motion sickness symptoms in patients with migrainous vertigo. These pilot data suggest a possible role for serotonin in the development of motion sickness symptoms in migraineurs with migrainous vertigo. PMID- 16369475 TI - Recent advances in the pathological basis and experimental management of impaired wound healing due to total-body irradiation. AB - Combined radiation and wound injury (CRWI) occurs in nuclear attacks and severe nuclear accidents. The possibility of radiological terrorist attack further emphasizes the significance of studies on CRWI. This kind of skin wound is very complex and difficult to heal since a high dose of total-body irradiation could delay wound healing and cause bone marrow dysplasia. Since the 1990s, the study of impaired wound healing due to total-body irradiation (TBI) has been emphasized in China. In this article, the pathological basis of the wound-healing process after TBI are reviewed and experimental management using traditional agents, growth factors, stem cells, and allo-skin transplantation in this kind of healing impaired wound is also discussed. PMID- 16369476 TI - ASD--lessons on genetic background from transgenic mice with inactive gene encoding metalloprotease, Tolloid-like 1 (TLL1). AB - Congenital heart defects still constitute serious medical problems. The background of such defects, however, is poorly understood. Little is also known about their pattern of inheritance. Lack of atrial and ventricular septum closures counts for a significant percentage of all congenital heart defects. Individuals harboring such defects always suffer poor quality of life. The only way to help those patients is cardiac surgery. Several genes have been implicated in the process of controlling heart development. Recent studies on transgenic mice null for tolloid-like 1 (tll1) gene revealed factors possibly involved in signaling pathways that, when absent, might affect heart development and cause problems mimicking those observed in atrial septal defects (ASDs). Lack of activity of this metalloprotease caused incomplete closure of the septum in murine fetal hearts and led to death in mid-gestation due to severe circulation problems. tll1 gene is located on the long arm of chromosome 4 (4q32-q33). The gene comprises of 6654 nucleotides and it encodes a protein 1013 amino acids long. Its human homologue tll1 is poorly investigated and until now no particular disorders have been linked to mutations in this gene. PMID- 16369477 TI - Non-alcoholic fatty liver disease and metabolic syndrome: what we know and what we don't know. AB - NAFLD is a very common asymptomatic liver condition that may progress to cirrhosis and hepatocellular carcinoma, and a relation to the different components of the metabolic syndrome has been found. In this review we highlight some of the epidemiological aspects of the two disorders and discuss some of the possible mechanisms and questions to be answered concerning the risk factors for the progression of this condition, as well as the need for more studies to focus on possible modalities of treatment. PMID- 16369478 TI - Lysophosphatidylcholine and secretory phospholipase A2 in vascular disease: mediators of endothelial dysfunction and atherosclerosis. AB - Lysophosphatidylcholine (LPC) is the major component of oxidized low density lipoprotein (oxLDL) and it has the ability to initiate or amplify several steps in atherogenesis due to its ability to impair endothelium-dependent vasorelaxation, enhance endothelial proliferation and permeability, stimulate adhesion and activation of lymphocytes, initiate chemotaxis of macrophages, impair migration and proliferation in vascular smooth muscle cells (SMCs), and modify platelet aggregation and coagulation pathways. For many of the LPC-induced effects, protein kinase C-dependent pathways have been implicated. In addition, modulation of ion current activity in the cell membrane, binding to a specific oxLDL receptor or to G-protein coupled receptors, as well as amplification of a highly oxidative state have all been postulated as likely mediating mechanisms. Secretory phopholipase A(2)-II (sPLA(2)-II) is one of the enzymes responsible for LPC production. sPLA(2)-II has been recently recognized as an independent risk factor for coronary artery disease. sPLA(2)-II favors the formation of bioactive lipids, stimulates SMC proliferation, activates macrophages enhancing lipid core formation and cytokine secretion, and binds to proteoglycans in the vessel wall matrix promoting lipid fusion and accumulation. The non-catalytic atherogenic effects of sPLA(2)-II are thought to be related to binding to an M-type receptor. Commonly used medications have been shown to decrease sPLA(2)-II activity generating a legitimate interest in the effects of the sPLA(2)-II pharmacologic antagonism. LPC and sPLA(2)-II are two very important mediators in atherosclerosis. Further research is warranted to clarify the cellular and molecular mechanisms that underlie their actions and to correlate in vitro data with clinical observations. PMID- 16369479 TI - Compound salvia pellet, a traditional Chinese medicine, for the treatment of chronic stable angina pectoris compared with nitrates: a meta-analysis. AB - BACKGROUND: Compound salvia pellet (CSP), consisting of active herbal ingredients extracted from Danshen (salvia miltiorrhiza), Sanqi (panax notoginseng), and Borneol (Cinnamomum camphora), is taken most frequently by patients with angina pectoris in China. To determine the efficacy and safety of CSP for the treatment of stable angina pectoris (SAP), a meta-analysis was undertaken. MATERIAL/METHODS: An extensive search including MEDLINE, EMBASE, BA, Chinese Biomedical Database (CBM), and Chinese Cochrane Centre Controlled Trials Register from 1994 to 2004 was performed. Data were extracted independently from the included trials by two reviewers. Statistical software (RevMan 4.1) provided by the Cochrane Collaboration was applied. RESULTS: Twenty-seven randomized controlled trials (RCTs) (n=3722), regardless of language or publication status, were identified. Generally, the methodological quality of the trials, assessed by the Jadad scale, was low, except for one with 3 points. Statistical pooling of the results showed that, compared with nitrates, CSP treatment had significant effect on the improvement of angina symptoms (RR=1.13, 95%CI=[1.07, 1.20]), showed greater increased effect on the improvement of electrocardiogram (ECG) results (RR=1.39, 95%CI=[1.28, 1.50]), and the percentage of patients with adverse events was significantly decreased in the treatment of CSP in comparison with nitrates (2.4% vs. 29.7%). CONCLUSIONS: CSP has significant effect on the improvement of angina symptoms and ECG results with few adverse events. However, the methodological quality of clinical trials with CSP for SAP needs to be improved, and the outcome measures should include mortality, quality of life, and other end-points. PMID- 16369480 TI - Calcium-dependent interaction of Lis1 with IQGAP1 and Cdc42 promotes neuronal motility. AB - Lis1 gene defects impair neuronal migration, causing the severe human brain malformation lissencephaly. Although much is known about its interactions with microtubules, microtubule-binding proteins such as CLIP-170, and with the dynein motor complex, the response of Lis1 to neuronal motility signals has not been elucidated. Lis1 deficiency is associated with deregulation of the Rho-family GTPases Cdc42, Rac1 and RhoA, and ensuing actin cytoskeletal defects, but the link between Lis1 and Rho GTPases remains unclear. We report here that calcium influx enhances neuronal motility through Lis1-dependent regulation of Rho GTPases. Lis1 promotes Cdc42 activation through interaction with the calcium sensitive GTPase scaffolding protein IQGAP1, maintaining the perimembrane localization of IQGAP1 and CLIP170 and thereby tethering microtubule ends to the cortical actin cytoskeleton. Lis1 thus is a key component of neuronal motility signal transduction that regulates the cytoskeleton by complexing with IQGAP1, active Cdc42 and CLIP-170 upon calcium influx. PMID- 16369481 TI - Molecular taxonomy of major neuronal classes in the adult mouse forebrain. AB - Identifying the neuronal cell types that comprise the mammalian forebrain is a central unsolved problem in neuroscience. Global gene expression profiles offer a potentially unbiased way to assess functional relationships between neurons. Here, we carried out microarray analysis of 12 populations of neurons in the adult mouse forebrain. Five of these populations were chosen from cingulate cortex and included several subtypes of GABAergic interneurons and pyramidal neurons. The remaining seven were derived from the somatosensory cortex, hippocampus, amygdala and thalamus. Using these expression profiles, we were able to construct a taxonomic tree that reflected the expected major relationships between these populations, such as the distinction between cortical interneurons and projection neurons. The taxonomic tree indicated highly heterogeneous gene expression even within a single region. This dataset should be useful for the classification of unknown neuronal subtypes, the investigation of specifically expressed genes and the genetic manipulation of specific neuronal circuit elements. PMID- 16369482 TI - DWnt4 regulates the dorsoventral specificity of retinal projections in the Drosophila melanogaster visual system. AB - In Drosophila melanogaster, the axons of retinal photoreceptor cells extend to the first optic ganglion, the lamina, forming a topographic representation. Here we show that DWnt4, a secreted protein of the Wnt family, is the ventral cue for the lamina. In DWnt4 mutants, ventral retinal axons misprojected to the dorsal lamina. DWnt4 was normally expressed in the ventral half of the developing lamina and DWnt4 protein was detected along ventral retinal axons. Dfrizzled2 and dishevelled, respectively, encode a receptor and a signaling molecule required for Wnt signaling. Mutations in both genes caused DWnt4-like defects, and both genes were autonomously required in the retina, suggesting a direct role of DWnt4 in retinal axon guidance. In contrast, iroquois homeobox genes are the dorsal cues for the retina. Dorsal axons accumulated DWnt4 and misprojected to the ventral lamina in iroquois mutants; the phenotype was suppressed in iroquois Dfrizzled2 mutants, suggesting that iroquois may attenuate the competence of Dfrizzled2 to respond to DWnt4. PMID- 16369483 TI - Chromogranin-mediated secretion of mutant superoxide dismutase proteins linked to amyotrophic lateral sclerosis. AB - Here we report that chromogranins, components of neurosecretory vesicles, interact with mutant forms of superoxide dismutase (SOD1) that are linked to amyotrophic lateral sclerosis (ALS), but not with wild-type SOD1. This interaction was confirmed by yeast two-hybrid screen and by co immunoprecipitation assays using either lysates from Neuro2a cells coexpressing chromogranins and SOD1 mutants or lysates from spinal cord of ALS mice. Confocal and immunoelectron microscopy revealed a partial colocalization of mutant SOD1 with chromogranins in spinal cord of ALS mice. Mutant SOD1 was also found in immuno-isolated trans-Golgi network and in microsome preparations, suggesting that it can be secreted. Indeed we report evidence that chromogranins may act as chaperone-like proteins to promote secretion of SOD1 mutants. From these results, and our finding that extracellular mutant SOD1 can trigger microgliosis and neuronal death, we propose a new ALS pathogenic model based on the toxicity of secreted SOD1 mutants. PMID- 16369484 TI - Modulation of microRNA processing and expression through RNA editing by ADAR deaminases. AB - Adenosine deaminases acting on RNA (ADARs) are involved in editing of adenosine residues to inosine in double-stranded RNA (dsRNA). Although this editing recodes and alters functions of several mammalian genes, its most common targets are noncoding repeat sequences, indicating the involvement of this editing system in currently unknown functions other than recoding of protein sequences. Here we show that specific adenosine residues of certain microRNA (miRNA) precursors are edited by ADAR1 and ADAR2. Editing of pri-miR-142, the precursor of miRNA-142, expressed in hematopoietic tissues, resulted in suppression of its processing by Drosha. The edited pri-miR-142 was degraded by Tudor-SN, a component of RISC and also a ribonuclease specific to inosine-containing dsRNAs. Consequently, mature miRNA-142 expression levels increased substantially in ADAR1 null or ADAR2 null mice. Our results demonstrate a new function of RNA editing in the control of miRNA biogenesis. PMID- 16369485 TI - Structure of Saccharomyces cerevisiae DNA polymerase epsilon by cryo-electron microscopy. AB - The structure of the multisubunit yeast DNA polymerase epsilon (Pol epsilon) was determined to 20-A resolution using cryo-EM and single-particle image analysis. A globular domain comprising the catalytic Pol2 subunit is flexibly connected to an extended structure formed by subunits Dpb2, Dpb3 and Dpb4. Consistent with the reported involvement of the latter in interaction with nucleic acids, the Dpb portion of the structure directly faces a single cleft in the Pol2 subunit that seems wide enough to accommodate double-stranded DNA. Primer-extension experiments reveal that Pol epsilon processivity requires a minimum length of primer-template duplex that corresponds to the dimensions of the extended Dpb structure. Together, these observations suggest a mechanism for interaction of Pol epsilon with DNA that might explain how the structure of the enzyme contributes to its intrinsic processivity. PMID- 16369486 TI - Autoproteolysis coupled to protein folding in the SEA domain of the membrane bound MUC1 mucin. AB - The single cell layer of the lungs and the gastrointestinal tract is protected by the mucus formed by large glycoproteins called mucins. Transmembrane mucins typically contain 110-residue SEA domains located next to the membrane. These domains undergo post-translational cleavage between glycine and serine in a characteristic GSVVV sequence, but the two peptides remain tightly associated. We show that the SEA domain of the human MUC1 transmembrane mucin undergoes a novel type of autoproteolysis, which is catalyzed by conformational stress and the conserved serine hydroxyl. We propose that self-cleaving SEA domains have evolved to dissociate as a result of mechanical rather than chemical stress at the apical cell membrane and that this protects epithelial cells from rupture. We further suggest that the cell can register mechanical shear at the mucosal surface if the dissociation is signaled via loss of a SEA-binding protein. PMID- 16369487 TI - The SPRY domain of SSB-2 adopts a novel fold that presents conserved Par-4 binding residues. AB - The four mammalian SPRY domain-containing SOCS box proteins (SSB-1 to SSB-4) are characterized by a C-terminal SOCS box and a central SPRY domain. We have determined the first SPRY-domain structure, as part of SSB-2, by NMR. This domain adopts a novel fold consisting of a beta-sandwich structure formed by two four stranded antiparallel beta-sheets with a unique topology. We demonstrate that SSB 1, SSB-2 and SSB-4, but not SSB-3, bind prostate apoptosis response protein-4 (Par-4). Mutational analysis of SSB-2 loop regions identified conserved structural determinants for its interaction with Par-4 and the hepatocyte growth factor receptor, c-Met. Mutations in analogous loop regions of pyrin and midline 1 SPRY domains have been shown to cause Mediterranean fever and Opitz syndrome, respectively. Our findings provide a template for SPRY-domain structure and an insight into the mechanism of SPRY-protein interaction. PMID- 16369488 TI - Transgenic rescue from embryonic lethality and renal carcinogenesis in the Nihon rat model by introduction of a wild-type Bhd gene. AB - We recently reported that a germline insertion of a single nucleotide in the rat homologue of the human Birt-Hogg-Dube gene (BHD) gives rise to dominantly inherited cancer in the Nihon rat model. In this study, we constructed transgenic Nihon rats with introduction of a wild-type Bhd gene to ascertain whether suppression of the Nihon phenotype is possible. Rescue from embryonic lethality of mutant homozygotes (Nihon/Nihon) and suppression of renal carcinogenesis in heterozygotes (Nihon/+) were both observed, defining the germline Bhd mutation in the Nihon rat as an embryonal lethal and tumor predisposing mutation. This transgenic rescue system will be useful to analyse Bhd gene function, its relation to tumorigenesis in vivo, and genetic-environmental interactions in carcinogenesis. PMID- 16369489 TI - Protocadherin 15 (PCDH15): a new secreted isoform and a potential marker for NK/T cell lymphomas. AB - Natural killer cells are well known to play an important role in immune defense against tumor development and viral infections. To further characterize new functionally relevant structures in these cells, we studied a series of monoclonal antibodies that we have raised against the NK cell line YT. One of these antibodies previously described as AY19, recognizes a 85 kD surface glycoprotein. Here we report the identification of a new secreted isoform of protocadherin 15, PCDH15C, which represents a potential associated protein for p85. Importantly, whereas protocadherins are absent from the surface of normal hematopoietic cells, we describe, for the first time, that PCDH15 is expressed in cytotoxic tumor-derived T- and NK-cell lines as well as in biopsies of nasal NK/T cell lymphomas. PMID- 16369490 TI - Cellular ITAM-containing proteins are oncoproteins in nonhematopoietic cells. AB - Immunoreceptor tyrosine-based activation motifs (ITAMs) are involved in the transduction of signals necessary for activation, differentiation, and survival in hematopoietic cells. Several viruses have been shown to encode ITAM-containing transmembrane proteins. Although expression of these viral proteins has in some cases been shown to transform nonhematopoietic cells, a causal role for a functional ITAM in this process has not been elucidated. To examine the potential transforming properties of ITAM-containing proteins, a recombinant protein consisting of ITAM-containing cytoplasmic regions of the B-cell antigen receptor was expressed in immortalized murine mammary epithelial and fibroblast cells. Mammary epithelial cells expressing this construct exhibited depolarized morphology in three-dimensional cultures. This transformed phenotype was characterized by a loss of anchorage dependence and hallmarks of epithelial to mesenchymal transition. Fibroblasts expressing this ITAM construct also lost contact inhibition and anchorage dependence. The transformed phenotype seen in both cell types was abrogated upon tyrosine to phenylalanine substitutions of the ITAMs. Inhibition of Syk tyrosine kinase, which associates with the ITAM, also prevented cell transformation. Our results indicate that expression of a nonviral ITAM-containing protein is sufficient for cell transformation. Despite lacking intrinsic enzymatic activity, ITAM-containing proteins can function as potent oncoproteins by scaffolding downstream mediators. PMID- 16369491 TI - Identification of putative oncogenes in lung adenocarcinoma by a comprehensive functional genomic approach. AB - Amplification and overexpression of putative oncogenes confer growth advantages for tumor development. We used a functional genomic approach that integrated simultaneous genomic and transcript microarray, proteomics, and tissue microarray analyses to directly identify putative oncogenes in lung adenocarcinoma. We first identified 183 genes with increases in both genomic copy number and transcript in six lung adenocarcinoma cell lines. Next, we used two-dimensional polyacrylamide gel electrophoresis and mass spectrometry to identify 42 proteins that were overexpressed in the cancer cells relative to normal cells. Comparing the 183 genes with the 42 proteins, we identified four genes - PRDX1, EEF1A2, CALR, and KCIP-1 - in which elevated protein expression correlated with both increased DNA copy number and increased transcript levels (all r > 0.84, two-sided P < 0.05). These findings were validated by Southern, Northern, and Western blotting. Specific inhibition of EEF1A2 and KCIP-1 expression with siRNA in the four cell lines tested suppressed proliferation and induced apoptosis. Parallel fluorescence in situ hybridization and immunohistochemical analyses of EEF1A2 and KCIP-1 in tissue microarrays from patients with lung adenocarcinoma showed that gene amplification was associated with high protein expression for both genes and that protein overexpression was related to tumor grade, disease stage, Ki-67 expression, and a shorter survival of patients. The amplification of EEF1A2 and KCIP-1 and the presence of overexpressed protein in tumor samples strongly suggest that these genes could be oncogenes and hence potential targets for diagnosis and therapy in lung adenocarcinoma. PMID- 16369492 TI - Repair and mutagenesis at oxidized DNA lesions in the developing brain of wild type and Ogg1-/- mice. AB - OGG1 (8-oxoguanine DNA glycosylase-1) is one of the main DNA glycosylases present in mammalian cells. The enzyme removes 7,8-dihydro-8-oxoguanine (8-oxoG) lesions, believed to be the most important oxidized lesions due to their relatively high incidence and their miscoding properties. This study shows that in prenatal mice brains the repair capacity for 8-oxoG is 5-10-fold higher than in adult mice brains. Western blot analysis and repair activity in extracts from Ogg1(-/-) mice revealed that OGG1 was responsible for the efficient 8-oxoG removal from prenatal mice. To investigate how OGG1 protects against oxidative stress-induced mutagenesis, pregnant Big Blue/wild-type and Big Blue/Ogg1(-/-) mice were exposed to nontoxic doses of gamma radiation. A 2.5-fold increase in the mutation frequency in Ogg1(-/-) mouse brains was obtained by exposure to 3.5 Gy at day 19 postfertilization. This was largely due to GC to TA transversions, believed to originate from 8-oxoG mispairing with A during replication. Furthermore, rapid cell divisions seemed to be required for fixation of mutations, as a similar dose of radiation did not increase the mutation frequency, or the frequency of GC to TA transversion, in the adult brain. PMID- 16369493 TI - Induction of tetraploidy through loss of p53 and upregulation of Plk1 by human papillomavirus type-16 E6. AB - Cancer cells are insensitive to many signals that inhibit growth of untransformed cells. Here, we show that primary human epithelial cells expressing human papillomavirus (HPV) type-16 E6/E7 bypass arrest caused by the DNA-damaging drug adriamycin and become tetraploid. To determine the contribution of E6 in the context of E7 to the resistance of arrest and induction of tetraploidy, we used an E6 mutant unable to degrade p53 or RNAi targeting p53 for knockdown. The E6 mutant fails to generate tetraploidy; however, the presence of E7 is sufficient to bypass arrest while the p53 RNAi permits both arrest insensitivity and tetraploidy. We published previously that polo-like kinase 1 (Plk1) is upregulated in E6/E7-expressing cells. We observe here that abnormal expression of Plk1 protein correlates with tetraploidy. Using the p53 binding-defective mutant of E6 and p53 RNAi, we show that p53 represses Plk1, suggesting that loss of p53 results in tetraploidy through upregulation of Plk1. Consistent with this hypothesis, overexpression of Plk1 in cells generates tetraploidy but does not confer resistance to arrest. These results support a model for transformation caused by HPV-16 where bypass of arrest and tetraploidy are separable consequences of p53 loss with Plk1 required only for the latter effect. PMID- 16369494 TI - Hypoxia stimulates breast carcinoma cell invasion through MT1-MMP and MMP-2 activation. AB - The process of cancer cell invasion involves degradation of the extracellular matrix (ECM) by proteases, integrin adhesion and cell motility. The role of ECM degrading proteases on the hypoxia-induced invasion of breast carcinoma cells was investigated. Hypoxia markedly increased the invasion capacity of MDA-MB-231 and MDA-MB-435 breast carcinoma cell lines. Matrix metalloproteinase (MMP) inhibitors blocked the hypoxia-induced invasion, whereas other protease inhibitors had no effect. Antibodies or siRNAs blocking either membrane type-1 MMP (MT1-MMP) or MMP 2 were effective in reducing the hypoxia-induced invasion. Serum-free reconstitution experiments confirmed the involvement of the MT1-MMP/MMP-2/tissue inhibitor of metalloproteinase-2 complex in this hypoxia-induced response. Overexpression of MT1-MMP in a poorly invasive breast cancer cell line, T47-D, promoted hypoxia-induced invasion and MMP-2 activation. Cell surface accumulation and activation of MT1-MMP without apparent regulation at the mRNA or protein levels indicated a post-translational adaptive response to hypoxia. Inhibition of the small GTPase RhoA eliminated the hypoxia-induced invasion and blocked the localization of MT1-MMP to the plasma membrane. Zymographic and molecular analysis of human breast tumors showed a strong correlation between hypoxic microenvironments and MMP-2 activation without changes in MT1-MMP expression. Our studies suggest that hypoxic tumor microenvironments promote breast cancer invasion through an MT1-MMP-dependent mechanism. PMID- 16369495 TI - Short-term induction and long-term suppression of HPV16 oncogene silencing by RNA interference in cervical cancer cells. AB - RNA interference-mediated gene silencing has the potential to block gene expression. A synthetic double-stranded small interfering RNA (siRNA) based on a sequence motif of 21 nucleotides from human papillomavirus 16 (HPV16) E6E7 bicistronic RNA was found to be a potent siRNA that suppresses expression of both the E6 and E7 oncogenes in HPV16+ CaSki and SiHa cells. When stably expressed as a short hairpin RNA in these cells, however, siRNA silencing of E6 and E7 expression was efficient only at early cell passages, but became inefficient with increased cell passages despite the continued expression of the siRNA at the same level. The loss of the siRNA function was duplicable in stable p53 siRNA cells, but not in stable lamin A/C siRNA cells, suggesting that it is gene selective. The cells resistant to siRNA function retained normal siRNA processing, duplex unwinding and degradation of the unwound sense strand and RNA-induced silencing complex formation, suggesting that loss of the siRNA function occurred at a later step. Surprisingly, the siRNA-resistant cells were found to express notably a cytoplasmic protein of approximately 50 kDa that specifically and characteristically interacted with the unwound, antisense strand E7 siRNA. Altogether, our data indicate that a potent siRNA targeting to an essential or regulatory gene might induce a cell to develop siRNA-suppressive function. PMID- 16369496 TI - DBC1 re-expression alters the expression of multiple components of the plasminogen pathway. AB - Deleted in bladder cancer 1 (DBC1) is a candidate gene for the bladder tumour suppressor locus at 9q33.1. The function of the gene is currently unknown but a cross-species sequence comparison suggests an important role, as it is highly evolutionarily conserved. Here, we transfected a nonexpressing human bladder cancer cell line with a set of human DBC1 cDNA constructs. The effect on global expression patterns was assessed using cDNA microarrays. The cell clone with the lowest level of DBC1 expression showed induced expression of 26 genes including plasminogen activator inhibitor 2 (SERPINB5; 4.6-fold), heparin-binding EGF-like growth factor precursor (DTR; 4.2-fold), small proline-rich protein 2B (SPRR2B; 3.6-fold), metallothionein 1 isoforms (MT1B/MT1A/MT-1F; from 2.9- to 3.2-fold), tissue-type plasminogen activator precursor (PLAT; 2.8-fold) and urokinase-type plasminogen activator precursor (PLAU; 2.7-fold). In clustering analysis, both PLAT and PLAU clustered with the functionally related urokinase plasminogen activator surface receptor (PLAUR; 1.9-fold). Furthermore, 14 human bladder tumours were analysed by real-time quantitative PCR using gene-specific primers for selected (n=20) genes. The expression levels of SERPINB5, PLAU, PLAUR and MT1 correlated with the DBC1 levels, suggesting previously unknown involvement of DBC1 in the urokinase-plasminogen pathway. PMID- 16369530 TI - APP locus duplication causes autosomal dominant early-onset Alzheimer disease with cerebral amyloid angiopathy. AB - We report duplication of the APP locus on chromosome 21 in five families with autosomal dominant early-onset Alzheimer disease (ADEOAD) and cerebral amyloid angiopathy (CAA). Among these families, the duplicated segments had a minimal size ranging from 0.58 to 6.37 Mb. Brains from individuals with APP duplication showed abundant parenchymal and vascular deposits of amyloid-beta peptides. Duplication of the APP locus, resulting in accumulation of amyloid-beta peptides, causes ADEOAD with CAA. PMID- 16369531 TI - Mutations in the CEL VNTR cause a syndrome of diabetes and pancreatic exocrine dysfunction. AB - Dysfunction of the exocrine pancreas is observed in diabetes, but links between concurrent exocrine and endocrine pancreatic disease and contributing genetic factors are poorly characterized. We studied two families with diabetes and exocrine pancreatic dysfunction by genetic, physiological and in vitro functional studies. A genome-wide screen in Family 1 linked diabetes to chromosome 9q34 (maximal lod score 5.07). Using fecal elastase deficiency as a marker of exocrine pancreatic dysfunction refined the critical chromosomal region to 1.16 Mb (maximal lod score 11.6). Here, we identified a single-base deletion in the variable number of tandem repeats (VNTR)-containing exon 11 of the carboxyl ester lipase (CEL) gene, a major component of pancreatic juice and responsible for the duodenal hydrolysis of cholesterol esters. Screening subjects with maturity-onset diabetes of the young identified Family 2, with another single-base deletion in CEL and a similar phenotype with beta-cell failure and pancreatic exocrine disease. The in vitro catalytic activities of wild-type and mutant CEL protein were comparable. The mutant enzyme was, however, less stable and secreted at a lower rate. Furthermore, we found some evidence for an association between common insertions in the CEL VNTR and exocrine dysfunction in a group of 182 unrelated subjects with diabetes (odds ratio 4.2 (1.6, 11.5)). Our findings link diabetes to the disrupted function of a lipase in the pancreatic acinar cells. PMID- 16369532 TI - Genome-wide transcription analyses in rice using tiling microarrays. AB - Sequencing and computational annotation revealed several features, including high gene numbers, unusual composition of the predicted genes and a large number of genes lacking homology to known genes, that distinguish the rice (Oryza sativa) genome from that of other fully sequenced model species. We report here a full genome transcription analysis of the indica rice subspecies using high-density oligonucleotide tiling microarrays. Our results provided expression data support for the existence of 35,970 (81.9%) annotated gene models and identified 5,464 unique transcribed intergenic regions that share similar compositional properties with the annotated exons and have significant homology to other plant proteins. Elucidating and mapping of all transcribed regions revealed an association between global transcription and cytological chromosome features, and an overall similarity of transcriptional activity between duplicated segments of the genome. Collectively, our results provide the first whole-genome transcription map useful for further understanding the rice genome. PMID- 16369533 TI - Nras loss induces metastatic conversion of Rb1-deficient neuroendocrine thyroid tumor. AB - Mutations in the gene encoding the retinoblastoma tumor suppressor predispose humans and mice to tumor development. Here we have assessed the effect of Nras loss on tumor development in Rb1 heterozygous mice. Loss of one or two Nras alleles is shown to significantly reduce the severity of pituitary tumors arising in Rb1(+/-) animals by enhancing their differentiation. By contrast, C-cell thyroid adenomas occurring in Rb1(+/-) mice progress to metastatic medullary carcinomas after loss of Nras. In Rb1(+/-)Nras(+/-) animals, distant medullary thyroid carcinoma metastases are associated with loss of the remaining wild-type Nras allele. Loss of Nras in Rb1-deficient C cells results in elevated Ras homolog family A (RhoA) activity, and this is causally linked to the invasiveness and metastatic behavior of these cells. These findings suggest that the loss of the proto-oncogene Nras in certain cellular contexts can promote malignant tumor progression. PMID- 16369534 TI - Homozygous L-SIGN (CLEC4M) plays a protective role in SARS coronavirus infection. AB - Severe acute respiratory syndrome (SARS) is caused by infection of a previously undescribed coronavirus (CoV). L-SIGN, encoded by CLEC4M (also known as CD209L), is a SARS-CoV binding receptor that has polymorphism in its extracellular neck region encoded by the tandem repeat domain in exon 4. Our genetic risk association study shows that individuals homozygous for CLEC4M tandem repeats are less susceptible to SARS infection. L-SIGN is expressed in both non-SARS and SARS CoV-infected lung. Compared with cells heterozygous for L-SIGN, cells homozygous for L-SIGN show higher binding capacity for SARS-CoV, higher proteasome-dependent viral degradation and a lower capacity for trans infection. Thus, homozygosity for L-SIGN plays a protective role during SARS infection. PMID- 16369535 TI - Intrinsic inhibition of transcription factor E2A by HLH proteins ABF-1 and Id2 mediates reprogramming of neoplastic B cells in Hodgkin lymphoma. AB - B cell differentiation is controlled by a complex network of lineage-restricted transcription factors. How perturbations to this network alter B cell fate remains poorly understood. Here we show that classical Hodgkin lymphoma tumor cells, which originate from mature B cells, have lost the B cell phenotype as a result of aberrant expression of transcriptional regulators. The B cell-specific transcription factor program was disrupted by overexpression of the helix-loop helix proteins ABF-1 and Id2. Both factors antagonized the function of the B cell determining transcription factor E2A. As a result, expression of genes specific to B cells was lost and expression of genes not normally associated with the B lineage was upregulated. These data demonstrate the plasticity of mature human lymphoid cells and offer an explanation for the unique classical Hodgkin lymphoma phenotype. PMID- 16369536 TI - Ablation of CD22 in ligand-deficient mice restores B cell receptor signaling. AB - CD22 is a negative regulator of B cell signaling, an activity modulated by its interaction with glycan ligands containing alpha2-6-linked sialic acids. B cells deficient in the enzyme (ST6Gal I) that forms the CD22 ligand show suppressed BCR signaling. Here we report that mice deficient in both CD22 and its ligand (Cd22-/ St6gal1-/- mice) showed restored B cell receptor (BCR) signaling, suggesting that the suppressed signaling of St6gal1-/- cells is mediated through CD22. Coincident with suppressed BCR signaling, B cells lacking ST6Gal I showed a net redistribution of the BCR to clathrin-rich microdomains containing most of the CD22, resulting in a twofold increase in the localization of CD22 together with the BCR. These studies suggest an important function for the CD22-ligand interaction in regulating BCR signaling and microdomain localization. PMID- 16369537 TI - Control of human immunodeficiency virus replication by cytotoxic T lymphocytes targeting subdominant epitopes. AB - Despite limited data supporting the superiority of dominant over subdominant responses, immunodominant epitopes represent the preferred vaccine candidates. To address the function of subdominant responses in human immunodeficiency virus infection, we analyzed cytotoxic T lymphocyte responses restricted by HLA-B*1503, a rare allele in a cohort infected with clade B, although common in one infected with clade C. HLA-B*1503 was associated with reduced viral loads in the clade B cohort but not the clade C cohort, although both shared the immunodominant response. Clade B viral control was associated with responses to several subdominant cytotoxic T lymphocyte epitopes, whereas their clade C variants were less well recognized. These data suggest that subdominant responses can contribute to in vivo viral control and that high HLA allele frequencies may drive the elimination of subdominant yet effective epitopes from circulating viral populations. PMID- 16369538 TI - A genetically encoded photosensitizer. AB - Photosensitizers are chromophores that generate reactive oxygen species (ROS) upon light irradiation. They are used for inactivation of specific proteins by chromophore-assisted light inactivation (CALI) and for light-induced cell killing in photodynamic therapy. Here we report a genetically encoded photosensitizer, which we call KillerRed, developed from the hydrozoan chromoprotein anm2CP, a homolog of green fluorescent protein (GFP). KillerRed generates ROS upon irradiation with green light. Whereas known photosensitizers must be added to living systems exogenously, KillerRed is fully genetically encoded. We demonstrate the utility of KillerRed for light-induced killing of Escherichia coli and eukaryotic cells and for inactivating fusions to beta-galactosidase and phospholipase Cdelta1 pleckstrin homology domain. PMID- 16369539 TI - Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli. AB - Bacterial protein secretion is important in the life cycles of most bacteria, in which it contributes to the formation of pili and flagella and makes available extracellular enzymes to digest polymers for nutritional purposes and toxins to kill host cells in infections of humans, animals and plants. It is generally accepted that nonpathogenic laboratory strains of Escherichia coli, particularly K12 strains, do not secrete proteins into the extracellular medium under routine growth conditions. In this study, we report that commonly used laboratory strains secrete YebF, a small (10.8 kDa in the native form), soluble endogenous protein into the medium, challenging the status quo view that laboratory strains do not secrete proteins to the medium. We further show that 'passenger' proteins linked to the carboxyl end of YebF are efficiently secreted. The function of YebF is unknown, but its use as a carrier for transgenic proteins provides a tool to circumvent toxicity and other contamination issues associated with protein production in E. coli. PMID- 16369540 TI - Erect leaves caused by brassinosteroid deficiency increase biomass production and grain yield in rice. AB - New cultivars with very erect leaves, which increase light capture for photosynthesis and nitrogen storage for grain filling, may have increased grain yields. Here we show that the erect leaf phenotype of a rice brassinosteroid deficient mutant, osdwarf4-1, is associated with enhanced grain yields under conditions of dense planting, even without extra fertilizer. Molecular and biochemical studies reveal that two different cytochrome P450s, CYP90B2/OsDWARF4 and CYP724B1/D11, function redundantly in C-22 hydroxylation, the rate-limiting step of brassinosteroid biosynthesis. Therefore, despite the central role of brassinosteroids in plant growth and development, mutation of OsDWARF4 alone causes only limited defects in brassinosteroid biosynthesis and plant morphology. These results suggest that regulated genetic modulation of brassinosteroid biosynthesis can improve crops without the negative environmental effects of fertilizers. PMID- 16369541 TI - Engineering and characterization of a superfolder green fluorescent protein. AB - Existing variants of green fluorescent protein (GFP) often misfold when expressed as fusions with other proteins. We have generated a robustly folded version of GFP, called 'superfolder' GFP, that folds well even when fused to poorly folded polypeptides. Compared to 'folding reporter' GFP, a folding-enhanced GFP containing the 'cycle-3' mutations and the 'enhanced GFP' mutations F64L and S65T, superfolder GFP shows improved tolerance of circular permutation, greater resistance to chemical denaturants and improved folding kinetics. The fluorescence of Escherichia coli cells expressing each of eighteen proteins from Pyrobaculum aerophilum as fusions with superfolder GFP was proportional to total protein expression. In contrast, fluorescence of folding reporter GFP fusion proteins was strongly correlated with the productive folding yield of the passenger protein. X-ray crystallographic structural analyses helped explain the enhanced folding of superfolder GFP relative to folding reporter GFP. PMID- 16369542 TI - Fluoroquinolone-modifying enzyme: a new adaptation of a common aminoglycoside acetyltransferase. AB - Antimicrobial-modifying resistance enzymes have traditionally been class specific, having coevolved with the antibiotics they inactivate. Fluoroquinolones, antimicrobial agents used extensively in medicine and agriculture, are synthetic and have been considered safe from naturally occurring antimicrobial-modifying enzymes. We describe reduced susceptibility to ciprofloxacin in clinical bacterial isolates conferred by a variant of the gene encoding aminoglycoside acetyltransferase AAC(6')-Ib. This enzyme reduces the activity of ciprofloxacin by N-acetylation at the amino nitrogen on its piperazinyl substituent. Although approximately 30 variants of this gene have been reported since 1986, the two base-pair changes responsible for the ciprofloxacin modification phenotype are unique to this variant, first reported in 2003 and now widely disseminated. An intense increase in the medical use of ciprofloxacin seems to have been accompanied by a notable development: a single function resistance enzyme has crossed class boundaries, and is now capable of enzymatically undermining two unrelated antimicrobial agents, one of them fully synthetic. PMID- 16369543 TI - A role for the scaffolding adapter GAB2 in breast cancer. AB - The scaffolding adapter GAB2 maps to a region (11q13-14) commonly amplified in human breast cancer, and is overexpressed in breast cancer cell lines and primary tumors, but its functional role in mammary carcinogenesis has remained unexplored. We found that overexpression of GAB2 (Grb2-associated binding protein 2) increases proliferation of MCF10A mammary cells in three-dimensional culture. Coexpression of GAB2 with antiapoptotic oncogenes causes lumenal filling, whereas coexpression with Neu (also known as ErbB2 and HER2) results in an invasive phenotype. These effects of GAB2 are mediated by hyperactivation of the Shp2-Erk pathway. Furthermore, overexpression of Gab2 potentiates, whereas deficiency of Gab2 ameliorates, Neu-evoked breast carcinogenesis in mice. Finally, GAB2 is amplified in some GAB2-overexpressing human breast tumors. Our data suggest that GAB2 may be a key gene within an 11q13 amplicon in human breast cancer and propose a role for overexpression of GAB2 in mammary carcinogenesis. Agents that target GAB2 or GAB2-dependent pathways may be useful for treating breast tumors that overexpress GAB2 or HER2 or both. PMID- 16369544 TI - Spying on cGMP with FRET. PMID- 16369545 TI - microRNA detection comes of age. PMID- 16369546 TI - Crafting new cages. PMID- 16369547 TI - The three 'C' s of chromosome conformation capture: controls, controls, controls. AB - Transcription regulation in higher eukaryotes is controlled by regulatory elements such as enhancers that are recognized by transcription factors. In many cases regulatory elements can be located at distances up to several megabases from their target genes. Recent evidence shows that long-range control of gene expression can be mediated through direct physical interactions between genes and these regulatory elements. Such looping interactions can be detected using the chromosome conformation capture (3C) methodology. Although 3C is experimentally straightforward, to draw meaningful conclusions one must carefully design 3C experiments and implement the conscientious use of controls. The general guidelines presented here should help experimental design and minimize misinterpretation of 3C experiments. PMID- 16369548 TI - Fluorescent sensors for rapid monitoring of intracellular cGMP. AB - Sensors based on fluorescence resonance energy transfer (FRET) are powerful tools to monitor signaling events in living mammalian cells. Here we describe development and use of new sensors for cyclic GMP (cGMP) based on cGMP binding domains from cGMP-dependent protein kinase I (GKI) and from phosphodiesterases (PDEs). The temporal and spatial resolution attained with the new sensors is superior to that of existing techniques, and permits direct recording and imaging of rapid cGMP-signaling events. PMID- 16369549 TI - In situ detection of miRNAs in animal embryos using LNA-modified oligonucleotide probes. AB - MicroRNAs (miRNAs) are 20-23 nucleotide (nt) RNA molecules that regulate gene expression post-transcriptionally. A key step toward understanding the function of the hundreds of miRNAs identified in animals is to determine their expression during development. Here we performed a detailed analysis of conditions for in situ detection of miRNAs in the zebrafish embryo using locked nucleic acid (LNA) modified DNA probes and report expression patterns for 15 miRNAs in the mouse embryo. PMID- 16369550 TI - Whole-genome genotyping with the single-base extension assay. AB - We describe an efficient, accurate and robust whole-genome genotyping (WGG) assay based on a two-color, single-base extension (SBE), single-nucleotide polymorphism (SNP)-scoring step. We report genotyping results for biallelic International HapMap quality control (QC) SNPs using a single probe per locus. We show scalability, throughput and accuracy of the system by resequencing homozygous loci from our 100k Human-1 Genotyping BeadChip. PMID- 16369551 TI - The nitrodibenzofuran chromophore: a new caging group for ultra-efficient photolysis in living cells. AB - Photochemical uncaging of bio-active molecules was introduced in 1977, but since then, there has been no substantial improvement in the properties of generic caging chromophores. We have developed a new chromophore, nitrodibenzofuran (NDBF) for ultra-efficient uncaging of second messengers inside cells. Photolysis of a NDBF derivative of EGTA (caged calcium) is about 16-160 times more efficient than photolysis of the most widely used caged compounds (the quantum yield of photolysis is 0.7 and the extinction coefficient is 18,400 M(-1) cm(-1)). Ultraviolet (UV)-laser photolysis of NDBF-EGTA:Ca(2+) rapidly released Ca(2+) (rate of 20,000 s(-1)) and initiated contraction of skinned guinea pig cardiac muscle. NDBF-EGTA has a two-photon cross-section of approximately 0.6 GM and two photon photolysis induced localized Ca(2+)-induced Ca(2+) release from the sarcoplasmic recticulum of intact cardiac myocytes. Thus, the NDBF chromophore has great promise as a generic and photochemically efficient protecting group for both one- and two-photon uncaging in living cells. PMID- 16369552 TI - A single-molecule method for the quantitation of microRNA gene expression. AB - MicroRNAs (miRNA) are short endogenous noncoding RNA molecules that regulate fundamental cellular processes such as cell differentiation, cell proliferation and apoptosis through modulation of gene expression. Critical to understanding the role of miRNAs in this regulation is a method to rapidly and accurately quantitate miRNA gene expression. Existing methods lack sensitivity, specificity and typically require upfront enrichment, ligation and/or amplification steps. The Direct miRNA assay hybridizes two spectrally distinguishable fluorescent locked nucleic acid (LNA)-DNA oligonucleotide probes to the miRNA of interest, and then tagged molecules are directly counted on a single-molecule detection instrument. In this study, we show the assay is sensitive to femtomolar concentrations of miRNA (500 fM), has a three-log linear dynamic range and is capable of distinguishing among miRNA family members. Using this technology, we quantified expression of 45 human miRNAs within 16 different tissues, yielding a quantitative differential expression profile that correlates and expands upon published results. PMID- 16369553 TI - Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy. AB - Lipid bodies have an important role in energy storage and lipid regulation. Here we show that lipid bodies are a major source of contrast in third-harmonic generation (THG) microscopy of cells and tissues. In hepatocytes, micrometer sized lipid bodies produce a THG signal 1-2 orders of magnitude larger than other structures, which allows one to image them with high specificity. THG microscopy with approximately 1,200 nm excitation can be used to follow the distribution of lipid bodies in a variety of unstained samples including insect embryos, plant seeds and intact mammalian tissue (liver, lung). We found that epi-THG imaging is possible in weakly absorbing tissues because bulk scattering redirects a substantial fraction of the forward-generated harmonic light toward the objective. Finally, we show that the combination of THG microscopy with two photon and second-harmonic imaging provides a new tool for exploring the interactions between lipid bodies, extracellular matrix and fluorescent compounds (vitamin A, NADH and others) in tissues. PMID- 16369554 TI - Strategies for protein coexpression in Escherichia coli. PMID- 16369555 TI - Get the balance right. PMID- 16369556 TI - Dampening inflammation. AB - This series of reviews addresses mechanisms whereby inflammation can be reduced, turned off or perhaps prevented, both in the context of natural dampening processes as well as possible approaches to anti-inflammatory therapy. Underlying the central issue is the implication that inflammation is indeed something inherently bad that needs to be dampened. Although this is certainly often the case and it can be argued that most human disease processes include some element of inflammation, nevertheless, inflammation is also natural, beneficial and, indeed, essential. This balance and its possible manipulation is a second theme running through the reviews. PMID- 16369557 TI - Immune cell migration in inflammation: present and future therapeutic targets. AB - The burgeoning field of leukocyte trafficking has created new and exciting opportunities in the clinic. Trafficking signals are being defined that finely control the movement of distinct subsets of immune cells into and out of specific tissues. Because the accumulation of leukocytes in tissues contributes to a wide variety of diseases, these 'molecular codes' have provided new targets for inhibiting tissue-specific inflammation, which have been confirmed in the clinic. However, immune cell migration is also critically important for the delivery of protective immune responses to tissues. Thus, the challenge for the future will be to identify the trafficking molecules that will most specifically inhibit the key subsets of cells that drive disease processes without affecting the migration and function of leukocytes required for protective immunity. PMID- 16369558 TI - Resolution of inflammation: the beginning programs the end. AB - Acute inflammation normally resolves by mechanisms that have remained somewhat elusive. Emerging evidence now suggests that an active, coordinated program of resolution initiates in the first few hours after an inflammatory response begins. After entering tissues, granulocytes promote the switch of arachidonic acid-derived prostaglandins and leukotrienes to lipoxins, which initiate the termination sequence. Neutrophil recruitment thus ceases and programmed death by apoptosis is engaged. These events coincide with the biosynthesis, from omega-3 polyunsaturated fatty acids, of resolvins and protectins, which critically shorten the period of neutrophil infiltration by initiating apoptosis. Consequently, apoptotic neutrophils undergo phagocytosis by macrophages, leading to neutrophil clearance and release of anti-inflammatory and reparative cytokines such as transforming growth factor-beta1. The anti-inflammatory program ends with the departure of macrophages through the lymphatics. Understanding these and further details of the mechanism required for inflammation resolution may underpin the development of drugs that can resolve inflammatory processes in directed and controlled ways. PMID- 16369559 TI - Limiting inflammatory responses during activation of innate immunity. AB - The idea of the importance of mounting an inflammatory response for effective immunity is supported by a multiplicity of experimental data. It is also well understood that resolution of inflammation is essential for maintaining the balance between health and disease. When the normal regulatory mechanisms are disturbed, the potential for developing chronic inflammatory diseases is increased. Inflammation is a key element in the response of the innate immune system to a variety of challenges, including those provided by bacterial and viral infection as well as by damaged or dying host cells. Here we review elements of innate immunity that lead to inflammation and some of the host mechanisms that allow for the resolution of the inflammatory responses. PMID- 16369561 TI - Breaking down the barriers to cancer immunotherapy. AB - Emerging insights into the mechanisms of activation and negative regulation of innate and adaptive immune cells are providing new opportunities for the development of safe and effective cancer vaccines. PMID- 16369562 TI - Pin1: a proline isomerase that makes you wheeze? PMID- 16369563 TI - Microclusters initiate and sustain T cell signaling. PMID- 16369564 TI - Signals for lymphocyte egress. PMID- 16369566 TI - Spotted-dick, a zinc-finger protein of Drosophila required for expression of Orc4 and S phase. AB - The highly condensed chromosomes and chromosome breaks in mitotic cells of a Drosophila mutant, spotted-dick/pita, are the consequence of defects in DNA replication. Reduction of levels of Spotted-dick protein, by either RNAi or mutation, leads to the accumulation of cells that have DNA content intermediate to 2N and 4N in proliferating tissues and also compromises endoreduplication in larval salivary glands. The Spotted-dick Zinc-finger protein is present in the nuclei of cells committed to proliferation but necessary in cells undertaking S phase. We show that Spotted-dick/Pita functions as a transcription factor and that, in cultured S2 cells, it is an activator of expression of some 30 genes that include the Orc4 gene, required for initiation of DNA replication. Chromatin immunoprecipitation indicates that it associates with the genes that it activates in S2 cells together with other sites that could represent genes activated in other tissues. We discuss the role of Spotted-dick in the coordination of cellular growth and DNA replication. PMID- 16369567 TI - The ATPase activity of MCM2-7 is dispensable for pre-RC assembly but is required for DNA unwinding. AB - Eukaryotes have six minichromosome maintenance (MCM) proteins that are essential for DNA replication. The contribution of ATPase activity of MCM complexes to their function in replication is poorly understood. We have established a cell free system competent for replication in which all MCM proteins are supplied by purified recombinant Xenopus MCM complexes. Recombinant MCM2-7 complex was able to assemble onto chromatin, load Cdc45 onto chromatin, and restore DNA replication in MCM-depleted extracts. Using mutational analysis in the Walker A motif of MCM6 and MCM7 of MCM2-7, we show that ATP binding and/or hydrolysis by MCM proteins is dispensable for chromatin loading and pre-replicative complex (pre-RC) assembly, but is required for origin unwinding during DNA replication. Moreover, this ATPase-deficient mutant complex did not support DNA replication in MCM-depleted extracts. Altogether, these results both demonstrate the ability of recombinant MCM proteins to perform all replication roles of MCM complexes, and further support the model that MCM2-7 is the replicative helicase. These data establish that mutations affecting the ATPase activity of the MCM complex uncouple its role in pre-RC assembly from DNA replication. PMID- 16369568 TI - The eloquent ape: genes, brains and the evolution of language. AB - The human capacity to acquire complex language seems to be without parallel in the natural world. The origins of this remarkable trait have long resisted adequate explanation, but advances in fields that range from molecular genetics to cognitive neuroscience offer new promise. Here we synthesize recent developments in linguistics, psychology and neuroimaging with progress in comparative genomics, gene-expression profiling and studies of developmental disorders. We argue that language should be viewed not as a wholesale innovation, but as a complex reconfiguration of ancestral systems that have been adapted in evolutionarily novel ways. PMID- 16369569 TI - The epigenetic progenitor origin of human cancer. AB - Cancer is widely perceived as a heterogeneous group of disorders with markedly different biological properties, which are caused by a series of clonally selected genetic changes in key tumour-suppressor genes and oncogenes. However, recent data suggest that cancer has a fundamentally common basis that is grounded in a polyclonal epigenetic disruption of stem/progenitor cells, mediated by 'tumour-progenitor genes'. Furthermore, tumour cell heterogeneity is due in part to epigenetic variation in progenitor cells, and epigenetic plasticity together with genetic lesions drives tumour progression. This crucial early role for epigenetic alterations in cancer is in addition to epigenetic alterations that can substitute for genetic variation later in tumour progression. Therefore, non neoplastic but epigenetically disrupted stem/progenitor cells might be a crucial target for cancer risk assessment and chemoprevention. PMID- 16369570 TI - Filtering transcriptional noise during development: concepts and mechanisms. AB - The assignation of cell fates during eukaryotic development relies on the coordinated and stable expression of cohorts of genes within cell populations. The precise and reproducible nature of this process is remarkable given that, at the single-cell level, the transcription of individual genes is associated with noise - random molecular fluctuations that create variability in the levels of gene expression within a cell population. Here we consider the implications of transcriptional noise for development and suggest the existence of molecular devices that are dedicated to filtering noise. On the basis of existing evidence, we propose that one such mechanism might depend on the Wnt signalling pathway. PMID- 16369571 TI - The role of double-strand break repair - insights from human genetics. AB - The efficient repair of DNA double-strand breaks is crucial in safeguarding the genomic integrity of organisms. Responses to double-strand breaks include complex signal-transduction, cell-cycle-checkpoint and repair pathways. Defects in these pathways lead to several human disorders with pleiotropic clinical features. Dissection of the molecular basis that underlies the diverse clinical features is enhancing our understanding of the damage-response mechanisms and their role in development, and might ultimately facilitate treatment. PMID- 16369572 TI - Microarray data analysis: from disarray to consolidation and consensus. AB - In just a few years, microarrays have gone from obscurity to being almost ubiquitous in biological research. At the same time, the statistical methodology for microarray analysis has progressed from simple visual assessments of results to a weekly deluge of papers that describe purportedly novel algorithms for analysing changes in gene expression. Although the many procedures that are available might be bewildering to biologists who wish to apply them, statistical geneticists are recognizing commonalities among the different methods. Many are special cases of more general models, and points of consensus are emerging about the general approaches that warrant use and elaboration. PMID- 16369573 TI - Genetic profiling of newborns: ethical and social issues. AB - Identifying genetic factors that could reliably predict health risks for individuals has the potential to bring great health benefits, both for the individuals concerned and for health-care providers. Genetic profiling at birth would allow a person's genome to be analysed at an early stage, and the data electronically stored for future use. However, although this might seem like an attractive proposition, it carries with it serious ethical and social concerns that would need to be addressed if the genetic profiling of newborns were ever to be considered on a population-wide basis. PMID- 16369574 TI - RNAi: a defensive RNA-silencing against viruses and transposable elements. AB - RNA silencing is a form of nucleic-acid-based immunity, targeting viruses and genomic repeated sequences. First documented in plants and invertebrate animals, this host defence has recently been identified in mammals. RNAi is viewed as a conserved ancient mechanism protecting genomes from nucleic acid invaders. However, these tamed sequences are known to occasionally escape this host surveillance and invade the genome of their host. This response is consistent with the overall idea that parasitic sequences compete with cells to systematically counter host defences. Using examples taken from the current literature, we illustrate the dynamic move-countermove game played between these two protagonists, the host cell and its parasitic sequences, and discuss the consequences of this game on genome stability. PMID- 16369575 TI - Endemic North African Quercus afares Pomel originates from hybridisation between two genetically very distant oak species (Q. suber L. and Q. canariensis Willd.): evidence from nuclear and cytoplasmic markers. AB - Hybridisation is a potent force in plant evolution, although there are few reported examples of stabilised species that have been created through homoploid hybridisation. We focus here on Quercus afares, an endemic North African species that combines morphological, physiological and ecological traits of both Q. suber and Q. canariensis, two phylogenetically distant species. These two species are sympatric with Q. afares over most of its distribution. We studied two Q. afares populations (one from Algeria and one from Tunisia), as well as several populations of both Q. suber and Q. canariensis sampled both within and outside areas where these species overlap with Q. afares. A genetic analysis was conducted using both nuclear (allozymes) and chloroplastic markers, which shows that Q. afares originates from a Q. suber x Q. canariensis hybridisation. At most loci, Q. afares predominantly possesses alleles from Q. suber, suggesting that the initial cross between Q. suber and Q. canariensis was followed by backcrossing with Q. suber. Other hypotheses that can account for this result, including genetic drift, gene silencing, gene conversion and selection, are discussed. A single Q. suber chlorotype was detected, and all Q. afares individuals displayed this chlorotype, indicating that Q. suber was the maternal parent. Q. afares is genetically, morphologically and ecologically differentiated from its parental species, and can therefore be considered as a stabilised hybrid species. PMID- 16369576 TI - High genetic diversity in cryptic populations of the migratory sutchi catfish Pangasianodon hypophthalmus in the Mekong River. AB - The detection and conservation of spawning units is of crucial importance in highly migratory species. The sutchi catfish Pangasianodon hypophthalmus (Pangasiidae; Teleostei) is a common large-sized tropical fish, which migrates annually to several upstream spawning sites on the Lower Mekong River and feeds on the huge floodplain of the Lower Mekong and Tonle Sap for the other half of the year. We hypothesised that because of the relative size of the feeding and spawning habitat, genetic variability would be high and homogeneous in foraging populations, but that spawning stocks would be distinct in space and time. To test these predictions, 567 individuals from 10 geographic locations separated by up to 1230 km along the Lower Mekong River were genotyped at seven microsatellite loci. The level of genetic diversity was much higher than other freshwater fish and reached values comparable to marine species (mean H(e)=0.757). All samples collected at the potential spawning sites deviated from Hardy-Weinberg expectations, suggesting admixture. Individual-based clustering methods revealed genetic heterogeneity and enabled the detection of three genetically distinct sympatric populations. There was no evidence of recent reduction in effective population size in any population. Contrasting with the vast extent of the feeding grounds, the shortage of spawning grounds seems to have moved sutchi catfish towards diachronous spawning. Hence the sustainable exploitation of this natural resource hinges on the conservation of the limited spawning grounds and open migration routes between the spawning and feeding grounds. PMID- 16369577 TI - Effect of variation in herkogamy on outcrossing within a population of Gilia achilleifolia. AB - The diverse features of floral morphology are often thought to be well-designed mechanisms to manipulate plant mating systems. We evaluated the effectiveness of one such mechanism, anther-stigma separation (herkogamy), in controlling variation in the level of outcrossing among plants in a population of Gilia achilleifolia. Variation in outcrossing rates within populations has the potential to influence the coevolution between inbreeding depression and mating system. Using four polymorphic allozymes, we compared the outcrossing-rate estimates of two groups of individuals under natural conditions: one group with low herkogamy and another with high herkogamy. The high herkogamy group had a higher outcrossing rate (0.572) than the low herkogamy group (0.335). This suggests that the within-population variation in outcrossing rate could potentially cause the previously observed association between herkogamy and inbreeding depression (Takebayashi and Delph, 2000). A previous study of floral traits among G. achilleifolia populations failed to detect a relationship between herkogamy and outcrossing rate, demonstrating that the functionality of traits may be obscured in among-population studies as a consequence of uncontrolled environmental variation. Additionally, the effect of herkogamy on outcrossing rate in delayed selfers such as G. achilleifolia may be particularly prominent when pollinator availability is low. Our population-level estimate of outcrossing rate (0.444) was somewhat lower than an estimate from the same population, 15 years prior to our study (0.75), suggesting that pollinator availability may fluctuate among years. Both within-year and among-year variation in the outcrossing rate may have a strong influence on mating-system evolution. PMID- 16369578 TI - Founder events as determinants of within-island and among-island genetic structure of Daphnia metapopulations. AB - The genetic structure of metapopulations offers insights into the genetic consequences of local extinction and recolonization. We studied allozyme variation in rock pool metapopulations of two species of waterfleas (Daphnia) with the aim to understand how these dynamics influence genetic differentiation. We screened 138 populations of D. magna and 65 populations of D. longispina from an area in the archipelago of southern Finland. The pools from which they were sampled are separated by distances between 1.5 and 4710 m and located on a total of 38 islands. The genetic population structure of the two species was strikingly similar, consistent with their similar metapopulation ecology. The mean F(PT) value (differentiation among pools with respect to the total metapopulation) was 0.55 and a hierarchical analysis showed that genetic differentiation was strong (>0.25) among pools within islands as well as among whole islands. Within islands, pairwise genetic differentiation increased with geographic distance, indicating isolation by distance due to spatially limited dispersal. Previous studies have shown strong founder events occurring during colonization in our metapopulation. We suggest that the genetic population structure in the studied metapopulations is largely explained by three consequences of these founder events: (i) strong drift during colonization, (ii) local inbreeding, which results in hybrid vigour and increased effective migration rates after subsequent immigration, and (iii) effects of selection through hitchhiking of neutral genes with linked loci under selection. PMID- 16369579 TI - Usefulness of catheter-directed thrombolysis using alteplase in peripheral vascular occlusion. AB - This study evaluated the safety and efficacy of alteplase in catheter-directed treatment for peripheral arterial and venous thrombosis and considered the optimal dosing regimen. Forty-four patients (49 encounters) underwent transcatheter therapy using alteplase between January and November 1999. The most common indications for thrombolysis were peripheral arterial occlusion (PAO) and venous thrombosis (38 patients, 43 encounters). Each encounter was reviewed for indication, dosage of alteplase, duration of infusion, concomitant use of anticoagulation, degree of lysis, and complications. Patients were divided into low-dose (0.5 to 1.0 mg/hr), mid-dose (1.0 to 1.5 mg/hr), and high-dose (>1.5 mg/hr) groups. For PAO, there was no significant difference in the success rate between the 3 dose groups. A lower complication rate was achieved in the low-dose group. For venous thrombosis, there was no difference in the overall success or complication rates for each of the 3 groups. Partial lysis was achieved more readily in the mid and high-dose groups, but the risk of serious complications was greater. Overall, the complete thrombolysis rate was 71% for PAO and 55% for venous thrombosis. Major and minor complication rates were 7% and 19%, respectively. An equivalent success rate with a lower complication rate can be achieved using a low-dose constant catheter-directed infusion of alteplase for cases of PAO. Cases of venous thrombosis had a lower overall success rate compared with PAO. A mid-dose infusion of alteplase can achieve greater complete and partial thrombolysis rates without increasing the complication rate. Major and minor complication rates were similar to the rates given in the published literature. PMID- 16369580 TI - Sarcoidosis: still a mystery? AB - Sarcoidosis is a systemic granulomatous disease of unknown etiology. It has diverse clinical manifestations, most frequently including pulmonary disorders. It is associated with immunological abnormalities, the intricacies of which have yet to be clearly delineated. In the immunologically susceptible individual, genetic, environmental, nutritional, and socioeconomic factors may play a governing role in its development. Sarcoidosis is a diagnosis of exclusion established by clinical manifestations, radiologic findings, and histologic evidence of noncaseating epithelioid-cell granulomas in >1 organ. We will discuss parameters that are helpful in making this diagnosis. PMID- 16369581 TI - Dexmedetomidine: a novel sedative-analgesic agent. AB - Since the first report of clonidine, an alpha2-adrenoceptor agonist, the indications for this class of drugs have continued to expand. In December 1999, dexmedetomidine was approved as the most recent agent in this group and was introduced into clinical practice as a short-term sedative (<24 hours). Alpha2 adrenoceptor agonists have several beneficial actions during the perioperative period. They decrease sympathetic tone, with attenuation of the neuroendocrine and hemodynamic responses to anesthesia and surgery; reduce anesthetic and opioid requirements; and cause sedation and analgesia. They allow psychomotoric function to be preserved while letting the patient rest comfortably. With this combination of effects, alpha2-adrenoceptor agonists may offer benefits in the prophylaxis and adjuvant treatment of perioperative myocardial ischemia. Furthermore, their role in pain management and regional anesthesia is expanding. Side effects consist of mild to moderate cardiovascular depression, with slight decreases in blood pressure and heart rate. The development of new, more selective alpha2 adrenoceptor agonists with improved side effect profiles may provide a new concept for the administration of perioperative anesthesia and analgesia. This review aims to give background information to improve understanding of the properties and applications of the novel alpha2-adrenoceptor agonist, dexmedetomidine. PMID- 16369583 TI - Medicine on the Internet. PMID- 16369582 TI - Use of the Mitek anchor in temporomandibular joint disc-repositioning surgery. PMID- 16369584 TI - A clinician's guide to resources on evidence-based medicine. PMID- 16369585 TI - Improving health care quality in Texas and the Baylor Health Care System. PMID- 16369587 TI - Boone Powell, Jr., MPH, FACHE: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369588 TI - Joel Tribble Allison, MS, FACHE: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369589 TI - Mark Timothy (Tim) Parris, MS: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369590 TI - Gary Dale Brock, MPH: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369591 TI - Acute pneumonia and systemic lupus erythematosus. PMID- 16369592 TI - In search of excellence--the Neonatal Intensive Care Quality Improvement Collaborative. AB - As part of its effort to improve the quality of care in the neonatal intensive care unit at Baylor University Medical Center (BUMC), the unit has participated in the Vermont Oxford Network. This network tracks outcomes and pools data, allowing comparisons and benchmarking. A group of 34 nurseries from the Vermont Oxford Network has collaborated in an innovative quality improvement initiative. This article describes this initiative, called the Neonatal Intensive Care Quality Collaborative 2000 project, and its impact on the neonatal service at BUMC. The project promotes the practice of 4 key habits: the habit for change, the habit for understanding the processes of care, the habit for collaborative learning, and the habit for using evidence-based practices of care. PMID- 16369594 TI - Abnormal pulmonary density discovered on routine chest radiograph. PMID- 16369595 TI - Hair loss and plaquelike skin lesions. PMID- 16369596 TI - Use of indicative and reciprocal electrocardiographic changes to help localize the site of coronary occlusion. PMID- 16369597 TI - Cross-reactivity of beta-lactam antibiotics. PMID- 16369598 TI - The physician's guide to medical malpractice. PMID- 16369599 TI - Facts and ideas from anywhere. PMID- 16369600 TI - Brugada syndrome. PMID- 16369601 TI - Depression, anxiety, and delirium in the terminally ill patient. PMID- 16369602 TI - Goals of care at the end of life. PMID- 16369603 TI - Conflict in the health care workplace. PMID- 16369604 TI - Two Oklahoma City disasters: the Oklahoma City bombing and the tornadoes--a blood bank perspective. PMID- 16369605 TI - Defining disaster: the emergency department perspective. PMID- 16369606 TI - Disaster prevention: lessons learned from the Titanic. PMID- 16369607 TI - Techniques for root cause analysis. PMID- 16369609 TI - Our dwindling national blood supply. PMID- 16369608 TI - Artificial blood: an update on current red cell and platelet substitutes. PMID- 16369610 TI - Otolaryngology at Baylor University Medical Center. PMID- 16369611 TI - Right ear pain and neck swelling. PMID- 16369612 TI - Leap for Life: innovative patient education to optimize outcomes among patients with cardiovascular disease. AB - OBJECTIVE: To evaluate the Leap for Life cardiovascular risk factor education program based on outcome measurements of self-reported hospital readmission, goal achievement, satisfaction, and educational model preference. SETTING: Four hospitals and one senior center of Baylor Health Care System in the Dallas, Texas, area. DESIGN: Administration of a satisfaction questionnaire and resurvey of participants by telephone at 3, 6, and 12 months. PATIENTS: 161 patients with cardiovascular disease enrolled in the Leap for Life program during calendar year 1997. Patients were primarily male (59%), with an average age of 66 years; they had an average of 4 cardiovascular disease risk factors. RESULTS: Of the 152 participants who took part in the telephone follow-up, 19% reported a hospital admission with a cardiovascular diagnosis during the 12-month follow-up period. Twenty-one percent reported meeting all of their goals; 56%, some of their goals; and 23%, none of their goals. All stated that the educational sessions met their informational needs, and 75% preferred the Leap for Life setting over other educational settings. CONCLUSION AND NEXT STEPS: These data provide initial validation of the program and are being used as a starting point for another assessment that involves individualized health enhancement measures and 6- and 12 month follow-up of participants using a survey designed to assess readiness, lifestyle changes, and quality of life. PMID- 16369614 TI - Chest discomfort and abnormal chest radiograph. PMID- 16369615 TI - Diffuse eruption of pigmented papules. PMID- 16369616 TI - Bigeminal rhythm. PMID- 16369617 TI - Recommendations for monitoring serum vancomycin concentrations. PMID- 16369618 TI - New federal standards for privacy of individual health information. PMID- 16369619 TI - Facts and ideas from anywhere. PMID- 16369620 TI - Preparing for bioterrorism. PMID- 16369621 TI - Provider and health care system response to a bioterrorist attack. PMID- 16369622 TI - Local perspectives on bioterrorism. An approach to terrorism preparedness: Parkland Health and Hospital System. PMID- 16369623 TI - Local perspectives on bioterrorism. Efforts of the Dallas-Fort Worth Hospital Council to prepare for bioterrorism. PMID- 16369624 TI - Local perspectives on bioterrorism. Emergency department preparedness at Baylor University Medical Center. PMID- 16369625 TI - Local perspectives on bioterrorism. Bioterrorism--a prospect for Dallas? PMID- 16369626 TI - Complex cardiac stab wound. PMID- 16369627 TI - Cardiac transplantation 40 years after a stab wound to the heart. PMID- 16369628 TI - Old unreduced posterior sternoclavicular dislocation and coracoid impingement. PMID- 16369629 TI - Idiopathic condylar resorption of the temporomandibular joint in teenage girls (cheerleaders syndrome). PMID- 16369630 TI - Orthopaedic surgery at Baylor University Medical Center. PMID- 16369631 TI - George Justice Race, MD, PhD, MSPH: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369632 TI - Severe anemia. PMID- 16369633 TI - Quality improvement efforts in the intensive care unit: development of a new heparin protocol. AB - CONTEXT: An ongoing process of identifying and implementing breakthrough projects to quickly improve patient outcomes and reduce costs in the intensive care unit. OBJECTIVE: To develop and test a new protocol for heparin administration in the intensive care unit, replacing the 5 existing protocols and ensuring compliance with the recommendations of the Fifth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy. DESIGN: A pilot study comparing results of 58 patients on the 5 existing protocols with those of 10 patients on the new heparin protocol. SETTING: Baylor University Medical Center, a large tertiary hospital in Dallas, Texas, with 115 adult intensive care unit beds. INTERVENTIONS: Heparin dosing was based on a patient's weight, and anticoagulation was monitored through the heparin assay. MAIN OUTCOME MEASURES: In the control group treated according to the existing protocols, 5 patients (8.6%) received optimal bolus doses, and 2 patients (3.4%) received optimal infusion doses. In the pilot group treated according to the new protocols, 9 patients (90%) received optimal bolus doses, and all received optimal infusion doses. RESULTS: The implementation of the new heparin protocol resulted in better patient care, improved nursing efficiency, and reduced costs. CONCLUSIONS: As shown in this example, a rapid breakthrough project following a plan-do-study-act cycle can apply best practices relatively quickly and lead to better patient outcomes. PMID- 16369636 TI - Dyspnea, cough, and fever for 7 days in an opiate addict. PMID- 16369637 TI - Pruritic eruption on the chest, arms, and buttocks. PMID- 16369638 TI - TV1 taller than TV6 as the only electrocardiographic indication of cardiac disease. PMID- 16369640 TI - Disclosure of personal information. PMID- 16369639 TI - A review of warfarin dosing and monitoring. PMID- 16369642 TI - Facts and ideas from anywhere. PMID- 16369643 TI - The Emergency Medical Treatment and Active Labor Act (EMTALA): what it is and what it means for physicians. PMID- 16369644 TI - The Health Insurance Portability and Accountability Act: is it really all that bad? PMID- 16369645 TI - Research compliance: entering Phase II. PMID- 16369646 TI - The role of spirituality in health care. PMID- 16369648 TI - The "missing link" in the chain of discovery of early melanoma of the skin. PMID- 16369649 TI - The Vikings and Baron Dupuytren's disease. PMID- 16369650 TI - History of nursing at Baylor University Medical Center. PMID- 16369647 TI - Neoplasms involving the heart, their simulators, and adverse consequences of their therapy. AB - Primary cardiac tumors involving the heart may be either benign or malignant. Most of the benign tumors are myxomas, which are most commonly located in the left atrium. Primary malignant neoplasms usually involve the myocardium and the interior of the cardiac cavities, whereas neoplasms metastatic to the heart most commonly involve pericardium, and pericardial effusion and constriction are the most common consequences. Computed tomography and magnetic resonance imaging are becoming the most useful instruments of precision for the diagnosis of cardiac tumors. Pericardial cysts, teratomas, lipomatous hypertrophy of the atrial septum, papillary fibroelastomas, thrombi, and sarcoid are frequently mistaken for cardiac neoplasms. There are a number of cardiac consequences of malignancy, including radiation heart disease, cardiac hemorrhages, cardiac infection, cardiac adiposity or the corticosteroid-treated heart, cardiac hemosiderosis, and toxicity due to anthracycline chemotherapy. PMID- 16369651 TI - Filipino nurses at Baylor University Medical Center: personal recollections. PMID- 16369652 TI - Michael Emmett, MD, MACP: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369653 TI - Marvin Jules Stone, MD, MACP: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16369654 TI - Baylor Health Care System quality initiatives: a view from the HealthTexas Provider Network. PMID- 16369656 TI - Headaches and back pain. PMID- 16369657 TI - Purplish, pruritic papules on the limbs. PMID- 16369659 TI - Utilizing causation. PMID- 16369658 TI - Chest pain and left bundle branch block. PMID- 16369660 TI - Facts and ideas from anywhere. PMID- 16369661 TI - [Treatment of chronic hepatitis C and molecular biology tests]. PMID- 16369662 TI - [The role of the third-generation oral cephalosporin cefditoren pivoxil in the treatment of community-acquired infection in adults]. PMID- 16369663 TI - [Streptococcus pneumoniae: antibiotic resistance and serotypes in a two-year period]. AB - Streptococcus pneumoniae is the main etiologic agent in community-acquired pneumonia, meningitis in adults and acute medium otitis in children. Our aim was to study antimicrobial susceptibility and serotypes of Streptococcus pneumoniae strains isolated in our medium. We evaluated 83 strains in a period of two years: 41% of the strains were invasive, and 47% of the strains were resistant to penicillin, 42.1% to erythromycin, 16.8% to cefotaxime, 34.9% to tetracycline and 16.8% to chloranphenicol. All studied strains were susceptible to vancomycin. We found 28 different serotypes. The most frequently found serotypes were 19, 6, 9, 23, 3, 15, and NT. The S. pneumoniae strains which showed the highest penicillin resistance belong to serotype 19. Multiresistant strains belong to serotypes 19, 6, 23 and NT. PMID- 16369664 TI - [Resistance of Mycobacterium tuberculosis in a four-year period in a Madrid hospital]. AB - Tuberculosis is considered a serious public health problem. Some factors, such as HIV infection and immigration, have had a major impact on the epidemiology of this illness in Spain. The problem has worsened in recent years due to the dissemination of multiresistant strains. Therefore, a periodic surveillance should be established with respect to the incidence and the resistances observed. In this study we collect M. tuberculosis isolates carried out in the years 2001, 2002, 2003 and 2004, and their susceptibility characteristics in patients from Area 2 in Madrid. To evaluate the isolates' susceptibilities, the MGIT 960 system was used. Of a total of 244 isolates, 15.2% were resistant to at least one antibiotic (different to streptomycin), and 29.9% of the isolates were obtained in samples from immigrant patients. In addition, the immigrant population affected showed a greater percentage of resistances (p <0.01) and a younger mean age (p <0.01) than the indigenous population. PMID- 16369665 TI - [Cytomegalovirus viral load and antigenemia in the monitorization of prophylactic treatment with valganciclovir in renal transplant recipients]. AB - Cytomegalovirus (CMV) disease is one of the most relevant infectious complications in solid organ transplant, and we must perform an appropriate prophylactic intervention. The goal of this study was to evaluate the effectiveness of prophylactic treatment with valganciclovir in renal transplant recipients in the first three months post transplantation by shell vial urine culture assay, and by measuring antigenemia (pp65) and CMV viral load, the latter by PCR. The population of the study included 100 renal transplant recipients. We analyzed the results of 36 patients recruited between November 2003 and July 2004 who were receiving a prophylactic oral treatment with valganciclovir, and who had finished the follow-up period of 90 days. The three tests mentioned above were performed on days 7, 15, 30, 45, 60, 75 and 90. No positive antigenemia was detected, the virus was cultured in a urine specimen and, in one patient, three measurements of viral load in serum were positive. Preliminary results of the study suggest that universal chemoprophylaxis with valganciclovir is effective for the prevention of CMV infection in renal transplant recipients and that, although all three tests used were useful, the measurement of CMV viral load seems to be the most appropriate method for monitoring these patients. PMID- 16369666 TI - [Pharmacokinetic/pharmacodynamic analysis to optimize antibacterial treatments: prediction of efficacy by using Montecarlo simulation techniques]. AB - The aim of this work is to provide a methodology to predict the potential efficacy of standard dosage schedules established for antimicrobials when used in clinical practice and administered to patients with different demographic characteristics. It is based on the application of pharmacokinetic and pharmacodynamic criteria (PK/PD analysis) to optimize dosification of this type of drug. Pharmacokinetic parameters such as the area under the plasma concentration-time curve (AUC) or maximum plasma concentration (Cmax) can be estimated from population kinetic models for each type of patient. Microbiological information, such as the MIC value, is also required. Using the above mentioned information and applying the Montecarlo simulation technique the probability of achieving the recommended value of a substituted variable related to efficacy may be estimated. The proposed methodology has been applied to levofloxacin when reportedly administered to patients showing different characteristics. The results reveal that this method allows us to know a priori whether or not the standard dosage is appropriate for a particular patient for whom the treatment is indicated. In summary, the proposed methodology provides us with a strategy for dosage individualization of antimicrobial agents that can be applied before initiating the treatment with no need for monitoring drug concentration, leading to an increase of clinical efficacy as well as a decreased risk of resistance development. PMID- 16369667 TI - [Implication of ermX genes in macrolide- and telithromycin-resistance in Corynebacterium jeikeium and Corynebacterium amycolatum]. AB - The activity of seven macrolides, clindamycin and telithromycin against clinical isolates of Corynebacterium spp. was studied. Of these, 36 isolates were identified as C. jeikeium and 57 as C. amycolatum. The frequency of resistance to erythromycin and other macrolides as well as clindamycin was high, with CMI(90) >256 microg/ml. Telithromycin showed the best activity, with 52.3% of C. amycolatum and 70% of C. jeikeium erythromycin-resistant strains susceptible to this ketolide. All strains had the MLSb constitutive phenotype. The ermX gene was present in all erythromycin-resistant strains, and in C. amycolatum was 100% homologous with that of C. striatum and C. diphtheriae. PMID- 16369668 TI - [Consumption of antibiotics in Spanish youth]. PMID- 16369669 TI - [Healthy young adult carriers of Streptococcus pneumoniae and antibiotic susceptibility]. PMID- 16369670 TI - [The awakening of the antibiotics era]. PMID- 16369671 TI - The unannounced standardised patient methodology--a potential feedback tool for registrar training. AB - BACKGROUND: The external clinical teaching visits (ECTV) remain the primary method of monitoring registrar progress and provide valuable feedback. The unannounced standardised patient (USP) methodology may represent an innovative tool to assess registrar performance and provide feedback that could complement the current ECTV program. OBJECTIVE: This article provides an overview of current feedback processes in registrar training, presents a review of the literature on the USP as an innovative performance assessment tool, and proposes its potential application for general practice registrar training as an effective audit and feedback tool. CONCLUSION: The USP methodology has been thoroughly tested, validated and embraced by many countries. Recently, the USP tool has been described as the validated, gold standard methodology to discriminate among variations in the quality of clinical practice. Most studies have used the USP tool to assess actual performance of practising doctors, with minimal attention being given to its potential as a feedback/teaching tool. The use of the USP for this purpose represents an opportunity for further research. PMID- 16369672 TI - Professional career needs of GPs and registrars working in northwestern NSW. AB - BACKGROUND: The Australian general practice workforce increasingly comprises international medical graduates. These doctors come from a range of professional backgrounds. AIM: This project aimed to document the current education, training and support needs of the general practice workforce, including international medical graduates, who currently train and/or practise in northwestern New South Wales. METHOD: Cross sectional survey of all 66 registrars and all 149 GPs who work in this area. RESULTS: Irrespective of doctors' origin or gender, priority information needs were related to indigenous health, rural health, and key community issues. Training and pre-employment opportunities, and continuing professional development were high priority areas for all participants. DISCUSSION: Educational initiatives are best targeted at identified groups but not necessarily reserved for international medical graduates only. Supporting the professional careers of doctors in a region requires the provision of integrated educational programs that focus on specific information and skills deficits, as well as requested programs. PMID- 16369673 TI - External clinical teaching visits in the regionalised environment. AB - BACKGROUND: The external clinical teaching (ECT) visit remains integral to the programs of all regional providers of the Australian General Practice Training (AGPT) program. METHOD: All 22 regional training providers (RTPs) were contacted by mail and telephone regarding their use of ECT visits. RESULTS: Responses were received from 17 of the 22 RTPs. There was considerable variation in approach to visitor training, reporting formats, provision of manuals, and the stated purpose of the visits. DISCUSSION: This study has provided a clearer picture of how ECT visits are functioning in the regionalised environment. There remains considerable variation in the use of these visits in Australian general practice training. New national guidelines promoting educational quality in ECT visits and protecting the diversity of regionalisation should be implemented. The use of these visits for performance appraisal needs further discussion. PMID- 16369674 TI - Three years of registrar rural exemptions--a review of a novel approach. AB - BACKGROUND: The Australian General Practice Training Program (AGPT) requires all registrars to undertake at least 6 months of training in rural areas. This can cause significant stress for general pathway registrars. Central to a fair, safe process for application of this policy is an equitable and humane rural exemption process. OBJECTIVE: WentWest's independent exemption assessment committee, consisting of an urban medical educator from another provider, a rural registrar liaison officer from another provider, and a lawyer, is described. OUTCOMES: The committee provides an independent, transparent and equitable process which protects the mentoring relationship between educators and registrars. Strict application of the AGPT criteria sometimes lacks compassion, suggesting the need for review of the criteria and some changes to committee function. DISCUSSION: The independent external committee may provide a model for decision making in other contentious areas of general practice training, such as remediation. The mandatory rural training policy requires reassessment 4 years after regionalisation, regarding its impact on workforce, learning and registrar wellbeing. PMID- 16369675 TI - Hybrid training for remotely situated general practice registrars--making best use of available opportunities. AB - BACKGROUND: Regional training providers (RTPs) working under the auspices of Australian General Practice Training (AGPT) sometimes encounter difficulties in delivering the AGPT program to general practice registrars, particularly those in the rural pathway. Registrars who have an employment bond with organisations such as state government health departments or the Australian Defence Forces are a prime example. The Remote Vocational Training Scheme (RVTS) presents an alternative model. OBJECTIVE: To develop, evaluate and improve a system which combines the best of the AGPT and RVTS delivery modes. DISCUSSION: The situations, arrangements and early outcomes from three AGPT and RVTS hybrid training arrangements in 2005 are described. Further formal evaluation will be necessary as the project progresses. PMID- 16369676 TI - A balancing act--the role of the general practice trainer. AB - BACKGROUND: The general practice trainer is recognised by The Royal Australian College of General Practitioners (the RACGP) as the most important element of Australian general practice training. The college has recently introduced new standards for trainers to ensure high quality general practice training. OBJECTIVE: This paper describes the various roles and responsibilities of the general practice trainer, and highlights the potential conflicts contained within them. DISCUSSION: The role of the general practice trainer is complex, challenging and diverse. It comprises separate--though interconnected--core roles of mentor, role model, clinical educator, assessor and pastoral carer. The balance of the various roles depends on the individual circumstances of the trainer-registrar relationship, and at times is potentially conflicting. Greater definition and consideration of the various facets of the role may lead to greater sustainability, satisfaction and reward for the trainer, and enhanced learning for the registrar. PMID- 16369677 TI - Progress in Aboriginal and Torres Strait Islander health training--an evaluation of the implementation of the GPET framework. AB - BACKGROUND: The General Practice Education and Training (GPET) board endorsed a framework for general practice training in Aboriginal and Torres Strait Islander health in September 2003. Training that conforms with the framework is required for accreditation of regional training providers. OBJECTIVE: This paper describes the first evaluation of the implementation of this framework by reporting on the extent to which seven key result areas (KRAs) of the framework have been addressed. DISCUSSION: The KRAs concerning delivery of Aboriginal and Torres Strait Islander training and recruitment and support of training posts were well achieved, while those concerning organisational and systems issues required further action. Good progress has been made towards implementation of the framework. Adequate funding and ongoing commitment to respectful and practical partnerships with the Aboriginal and Torres Strait Islander community controlled sector are required to further this. PMID- 16369678 TI - Peer learning, lectures and online learning--putting it all together. AB - INTRODUCTION: This paper evaluates usage of discussion forums forming part of an integrated online learning environment that supplements a face to face general practice registrar training program. METHOD: Basic general practice registrars were given training in the use of the online learning environment, which was explicitly linked to their other educational activities. A group of advanced registrars was given access to the online environment without any formal training or support and without specific curriculum linkage. Usage data were in the form of quantitative analysis of site logs, posts to discussion forums before and after a website redesign, and qualitative feedback from focus groups. RESULTS: The majority of basic registrars accessed forums to read messages. Only one third posted messages. The number of registrars to access forums was similar before and after site redesign. Log data indicated greater exploration of resources in the redesigned environment. The group of advanced registrars used the online resources significantly less than the basic registrars. DISCUSSION: These data underscore the importance of linking curriculum to online learning resources, and of providing adequate information technology training and support. PMID- 16369679 TI - General practice supervision at a distance--is it remotely possible? AB - BACKGROUND: The Remote Vocational Training Scheme (RVTS) trains GPs while in solo rural or remote practice. General practice registrars choose an experienced rural or remote practitioner as their distant supervisor. METHOD: During a teacher training telephone tutorial series RVTS general practice supervisors discussed the unique features of this model of general practice training, and shared their perspectives on how to make this distance supervision effective. RESULTS: This paper outlines the unique qualities of general practice registrars, the registrar supervisor relationship and the content of tutorials, in the context of remote supervision. This paper suggests ways to provide educational support despite geographical distance. DISCUSSION: Experienced rural and remote GPs can provide effective supervision at a distance. The RVTS provides an innovative, alternative model for training GPs for rural and remote practice. Further qualitative research on this topic is required. PMID- 16369680 TI - Addiction medicine--training for general practice registrars. AB - INTRODUCTION: Illicit drug use is a significant problem within Australia. However, GPs are reluctant to treat these problems due to a perceived lack of knowledge and confidence. METHOD: All advanced general practice registrars training through the Victorian Metropolitan Alliance in 2004 attended a 1 day pilot addiction medicine workshop, followed by completion of a questionnaire to assess the acceptability of the training and its impact on their knowledge and confidence. RESULTS: Forty-six general practice registrars attended the workshops with all but one (98%) completing the postworkshop questionnaire. More than 90% of registrars agreed that the material presented was relevant to their work and that they felt more confident dealing with addiction issues in practice. DISCUSSION: Addiction medicine training is relevant to the work of doctors training to become GPs and can increase knowledge and confidence. More extensive research is needed to assess whether this increase in knowledge and confidence is sustained over time and is translated into practice. PMID- 16369681 TI - Registrar attitudes to mental health care provision--does level one training make a difference? AB - INTRODUCTION: This study was designed to demonstrate whether compulsory level one mental health training leads to a change in knowledge, skills and attitudes for general practice registrars. METHODS: All registrars in the pilot level one course completed an attitudinal questionnaire and a case based check program before and after training. Registrars also completed a participant satisfaction instrument. RESULTS: All registrars showed a positive shift in attitudinal and knowledge indicators following training. The degree of change did not relate to their initial scores. DISCUSSION: Mental health provides a useful model of chronic and complex care. The three step model of assessment, planning and review can be generalised to other high prevalence illnesses. The provision of level one training addresses specific barriers to general practice mental health care and may influence registrar interest in continuing mental health education. Consideration should be given to making level one mental health training a compulsory component of vocational training. Evidence would be strengthened by examining registrars' clinical behaviour and patient outcomes before and after training. PMID- 16369682 TI - Learning needs analysis--developing a new tool for general practice training. AB - BACKGROUND: General practice registrars are a diverse group of doctors, each with a unique set of learning needs. This diversity sets training providers the challenge of identifying the learning needs of individual registrars early and then tailoring their educational programs to meet those needs. OBJECTIVE: This article describes the development of a multicomponent learning needs analysis tool designed to better identify registrars' learning needs in both knowledge content and attitudinal areas. DISCUSSION: The accurate identification of registrars' individual and common learning needs allows the creation of more appropriate learning plans and the tailoring of shared educational activities to suit both individuals and the group. PMID- 16369683 TI - Systems change in Australian general practice--early impact of the National Primary Care Collaboratives. AB - BACKGROUND: The capacity to translate research findings into clinical practice is a core skill for general practice registrars if we are to bridge the gap between evidence and practice, thus improving quality of care. OBJECTIVE: The National Primary Care Collaboratives approach is described as a way of improving health outcomes through systems change, more closely linking local practice with structures which have been proved to be effective elsewhere. The opportunity for general practice registrars to learn from and be involved in this approach is discussed. DISCUSSION: Competencies in systems based approaches and practice based learning are part of modern practice and should be embedded within the revised Royal Australian College of General Practitioners (the RACGP) curriculum. The National Primary Care Collaboratives Program is well placed to assist in the acquisition of these competencies. PMID- 16369684 TI - Teaching on the run--general practice training between consultations. AB - BACKGROUND: General practitioner teachers play a growing role in medical education. Much of the teaching is done during and between patient consultations, ie. 'on the run'. This presents challenges in terms of time available for teaching, teacher training and feedback on teacher performance. Australian Government funded programs have been developed to train clinical teachers in hospital settings; these might be adaptable to the general practice setting. OBJECTIVE: This article describes a project aimed to adapt current programs to the needs of GP teachers, and present them in workshops around Tasmania. DISCUSSION: The adapted program was well received by workshop participants, who reported significant increase in knowledge and skill in teaching, and a change in attitude to teaching in their practices. The program will be continued and expanded to encourage more GPs, general practice registrars and senior medical students to teach on the run. PMID- 16369685 TI - Context specific learning opportunities--adapting training to discrete areas. AB - BACKGROUND: Australian general practice training was regionalised in 2002 to better meet the specific health care needs of local communities. As a result, 22 unique regions were established, each containing a variety of training practices and learning contexts. The core learning opportunities in distinct learning contexts have not been well explored. METHODS: The research group identified criteria for delineating learning contexts and applied them to the training practices in the Northern Territory. Using focus groups of general practice registrars, we identified the learning opportunities available in each context. RESULTS: Context criteria were identified as: model of health service organisation and delivery; rurality; and community demography. Four distinct contexts were identified: mainstream general practice; regional hospital; Aboriginal Medical Service (AMS); and Aboriginal community clinic. Each learning context demonstrated both strengths and deficiencies, and also provided unique learning opportunities. DISCUSSION: Benefits in defining the learning contexts and opportunities within a general practice training region apply to registrars, trainers, providers, policy makers and standard setters. Context definition provides a valuable tool for more targeted and effective training, contributes to curriculum development and facilitates a more responsive and appropriate education program. We encourage adaptation of this model to other regions. PMID- 16369686 TI - Role of calcineurin in Porphyromonas gingivalis-induced myocardial cell hypertrophy and apoptosis. AB - BACKGROUND AND OBJECTIVE: Periodontal pathogen Porphyromonas gingivalis (P. gingivalis) increased cardiomyocyte hypertrophy and apoptosis whereas Actinobaeillus actinomycetemcomitans and Prevotella intermedia had no effects. The purpose of this study is to clarify the role of calcineurin signaling pathway in P. gingivalis-induced H9c2 myocardial cell hypertrophy and apoptosis. METHODS: DNA fragmentation, nuclear condensation, cellular morphology, calcineurin protein, Bcl2-associated death promoter (Bad) and nuclear factor of activated T cell (NFAT)-3 protein products in cultured H9c2 myocardial cell were measured by agarose gel electrophoresis, DAPI, immunofluorescence, and Western blotting following P. gingivalis and/or pre-administration of CsA (calcineurin inhibitors cyclosporin A). RESULTS: P. gingivalis not only increased calcineurin protein, NFAT-3 protein products and cellular hypertrophy, but also increased DNA fragmentation, nuclear condensation and Bad protein products in H9c2 cells. The increased cellular sizes, DNA fragmentation, nuclear condensation, and Bad of H9c2 cells treated with P. gingivalis were all significantly reduced after pre administration of CsA. CONCLUSION: Our findings suggest that the activity of calcineurin signal pathway may be initiated by P. gingivalis and further lead to cell hypertrophy and death in culture H9c2 myocardial cells. PMID- 16369687 TI - Quantification of S-adenosyl methionine in microbial cell extracts. AB - A sensitive method for quantification of S-adenosyl methionine (SAM) in microbial cell extracts was developed and applied to Corynebacterium glutamicum. The method is based on SAM being completely hydrolyzed into (18)O-homoserine when extracted in boiling H(2) (18)O and thus can be clearly distinguished by GC-MS analysis from naturally labeled homoserine present in the cell extract. Additional quantification of the total homoserine pool, representing both SAM and homoserine, via HPLC allows separate determination of both metabolites. PMID- 16369688 TI - Isolation of a fusion protein containing the antigenic domain 1 of human cytomegalovirus glycoprotein B and its application in ELISA tests. AB - The glycoprotein B (gB) of human cytomegalovirus represents a dominant antigen for the humoral immune response. The immunodominant region on gB is the antigenic domain 1 (AD-1), a complex structure that requires a minimal continuous sequence of more than 75 amino acids for antibody binding. In this study, this domain was expressed in Escherichia coli as a fusion protein with beta-galactosidase but yielded insoluble protein aggregates as inclusion bodies. To recover the fusion protein, inclusion bodies were solubilized by two extractions with urea 8 M: and the fusion protein then isolated using gel filtration chromatography. After confirmation of fusion protein antigenicity by Western blotting, the purified product was used as the capturing antigen in an enzyme-linked immunosorbent assay (ELISA) to determine the presence of viral antibodies in serum samples of pregnant women. A cut-off point of approximately 0.2 absorbance units could discriminate the results of seropositive from seronegative pregnant women. The data indicates the potential usefulness of the fusion protein for the development of immunoassay for detection of the HCMV antibodies. PMID- 16369689 TI - Selection of peptides for lipopolysaccharide binding on to epoxy beads and selective detection of Gram-negative bacteria. AB - Lipopolysaccharide (LPS)-binding peptides were enriched by using epoxy beads as a novel support to immobilize LPS for a phage displayed peptide library screening. The sequence of Phe-Ala-Pro-Trp (FAPW) was the most significant consensus motif of 10 selected clones, and Pro-Phe (PF) was the key dipeptide for binding at the apex of the loop to form a characteristic structure of CXXPFXXXC. Moreover, AWLPWAK, one of the highly conserved heptamer peptides, could detect specifically Gram-negative bacteria via a whole cell binding test at 10(6) cells ml(-1). PMID- 16369690 TI - Purification and characterization of a cold-active iron superoxide dismutase from a Psychrophilic Bacterium, Marinomonas sp. NJ522. AB - Marinomonas sp. NJ522, isolated from Antarctic sea ice, produces a cold-active iron superoxide dismutase (SOD; EC 1.15.1.1). The purified SOD was dimeric and had an approx. Mr of 48 kDa. Highest activity was detected from pH 8 to 10 and at 40 degrees C (assayed over 10 min). Activity at 0 degree C was nearly 35% of the maximum activity. PMID- 16369691 TI - Expression of galactose permease and pyruvate carboxylase in Escherichia coli ptsG mutant increases the growth rate and succinate yield under anaerobic conditions. AB - In Escherichia coli, disruption of ptsG, which encodes the glucose-specific permease of the phosphotransferase transport system (PTS) protein EIICB(Glc), is crucial for high succinate production. This mutation can, however, cause very slow growth and low glucose consumption rates. The ptsG mutant (TUQ2), from wild type E. coli W1485, and E. coli galP (encoding galactose permease) and glk (encoding glucose kinase) gene expression plasmids were constructed. TUQ2 increased the generation time to approximately 4 h and gave a higher final cell density of 0.5 g/l by expression of galP. However, glk expression had no effect on the mutant. After expression of pyruvate carboxylase (PYC) and galactose permease, the ptsG mutant showed higher succinate yield (1.2 mol/mol glucose) and the specific rate of glucose consumption from 0.33 to 0.6 g/1 h. PMID- 16369692 TI - Redirecting carbon flux in Torulopsis glabrata from pyruvate to alpha ketoglutaric acid by changing metabolic co-factors. AB - The nutrition conditions needed to redirect the carbon flux in Torulopsis glabrata, a pyruvate hyper-production yeast, from pyruvate to alpha-ketoglutaric acid (KG) were investigated in a stirred fermentor. A minor amount of KG (1.3 gl( 1)) was produced when NaOH was used to control the pH, while 12 g KG l(-1) was produced when CaCO(3) was used instead. When thiamine and biotin were included in the medium, 13 g KG l(-1) and 68 g pyruvate l(-1) were produced after 48 h when glucose was nearly consumed (approximately 5 gl(-1)). With fermentation continuing for a further 16 h, the concentration of pyruvate decreased to 31 gl( 1), and KG increased to 30 gl(-1). KG thus accumulated at the expense of pyruvate consumption. PMID- 16369693 TI - Genome-wide analysis of the expression profile of Saccharomyces cerevisiae in response to treatment with the plant isoflavone, wighteone, as a potential antifungal agent. AB - The limitations of currently available antifungal agents and the rapid emergence of drug-resistant strains necessitate more efficient approaches to screening and developing novel antifungal drugs. The antifungal activity of the natural products of a series of plants was evaluated and wighteone, 5, 7, 4'-trihydroxy-6 (gamma,gamma-dimethylallyl)isoflavone showed excellent anti-yeast activity (MIC against Saccharomyces cerevisiae was 4 microg/ml). Transcriptome profiling of wighteone-treated S. cerevisiae indicated that wighteone is different from commonly used antifungal compounds in its mode of action. PMID- 16369694 TI - Characterization of a bifunctional cytidine 5'-monophosphate N-acetylneuraminic acid synthetase cloned from Streptococcus agalactiae. AB - Recombinant CMP-sialic acid synthetase, cloned from Streptococcus agalactiae serotype V strain 2603 V/R, is bifunctional having both CMP-sialic acid synthetase and acetylhydrolase (acylesterase) activities. The enzyme is active over a wide pH range with an optimal CMP-sialic acid synthetase activity at pH 9.0 and an optimal acetylhydrolase activity at pH 8.0. A metal cofactor (either Mg(2+) or Mn(2+)) is required for the CMP-sialic acid synthetase activity but is not for acetylhydrolase activity. Both catalytic functions, however, are impaired by high concentrations of Mn(2+). PMID- 16369695 TI - Development of transformation system in Monascus purpureus using an autonomous replication vector with aureobasidin A resistance gene. AB - To enhance the variety of genetic tools and thus to promote molecular genetic study, aureobasidin A and its resistance gene were adopted as a new marker system together with the incorporation of the Gateway system to facilitate the introduction of long heterologous DNA fragments into Monascus purpureus. The minimum inhibitory concentration of aureobasidin A against Monascus was 0.05 microg/ml and a transformation efficiency of 17 colonies/microg DNA was obtained by the protoplast-PEG method with the vector pAUR316, containing the aureobasidin A resistance gene. Southern analysis of the transformants confirmed that pAUR316 exists as an independent vector, demonstrating that the AMA1 sequence acts as the autonomous replication sequence in M. purpureus. Through the use of the Gateway system, a polyketide synthase gene (7.8 kbp) responsible for citrinin biosynthesis was introduced. As a result, the transformants showed 1.5-fold higher production of citrinin than the wild-type strain. PMID- 16369696 TI - High level stable expression of pharmacologically active human M1-M5 muscarinic receptor subtypes in mammalian cells. AB - cDNAs encoding for five mAChR subtypes (M1-M5) were cloned under different promoters in various eukaryotic vectors and each subtype was expressed in different mammalian cell lines. CHO-K1 cell line was the best for generating stable cell lines expressing muscarinic receptors. Immunofluorescence and flow cytometry revealed that expression of M1-M5 was primarily localized on the cell membrane. Western blotting and radio-ligand binding studies revealed that expression of each receptor was stable at higher passages. PMID- 16369697 TI - Purification and properties of a beta-1,3-glucanase from Chaetomium sp. that is involved in mycoparasitism. AB - A beta-1,3-glucanase was detected, using laminarin as substrate, in the culture broth of Chaetomium sp. Major activity was associated with a 70 kDa protein band visualized on a polyacrylamide gel. beta-1,3-Glucanase was purified by a one step, native gel purification procedure. Optimal activity was observed at pH 6.0 and 30 degrees C (over 30 min). It could degrade cell walls of plant pathogens including Rhizoctonia solani, Gibberella zeae, Fusarium sp., Colletotrichum gloeosporioides and Phoma sp. The N-terminal amino acid residues of the purified beta-1,3-glucanase are PYQLQTP, which do not exhibit homology to other fungal beta-1,3-glucanases suggesting it may be a novel enzyme. PMID- 16369699 TI - Frontal electroencephalogram activation asymmetry, emotional intelligence, and externalizing behaviors in 10-year-old children. AB - The purpose of the present study was to examine the relations among resting frontal brain electrical activity (EEG) (hypothesized to reflect a predisposition to positive versusnegative affect and ability to regulate emotions), emotional intelligence, and externalizing behaviors in a sample of non-clinical 10-year-old children. We found that boys had significantly lower emotional intelligence than girls, and low emotional intelligence was associated with significantly more externalizing behaviors (i.e., aggression and delinquency), replicating previous work. We also found that children with higher reported externalizing behaviors exhibited significantly greater relative right frontal EEG activity at rest compared with children with little to no externalizing behavioral problems. There was, however, no relation between emotional intelligence and the pattern of resting frontal EEG activity. Thus, emotional intelligence and the pattern of frontal EEG activation at rest are independent predictors of externalizing behaviors in children. Findings also suggest that individual differences in emotional intelligence may not be based on differences in levels of emotional regulation or the generation of positive affect as reflected in frontal EEG asymmetries, but rather other social and cognitive competencies required for adaptive behavior. PMID- 16369700 TI - Reducing whole body physiologically based pharmacokinetic models using global sensitivity analysis: diazepam case study. AB - There are situations in drug development where one may wish to reduce the dimensionality and complexity of whole body physiologically based pharmacokinetic models. A technique for formal reduction of such models, based on global sensitivity analysis, is suggested. Using this approach mean and variance of tissue(s) and/or blood concentrations are preserved in the reduced models. Extended Fourier amplitude sensitivity test (FAST), a global sensitivity technique, takes a sampling approach, acknowledging parameter variability and uncertainty, to calculate the impact of parameters on concentration variance. We used existing literature rules for formal model reduction to identify all possible smaller dimensionally models. To discriminate among those competing mechanistic models extended FAST was used, whereby we treated model structural uncertainty as another factor contributing to the overall uncertainty. A previously developed 14 compartment whole body physiologically based model for diazepam disposition in rat was reduced to three alternative reduced models, with preserved arterial mean and variance concentration profiles. PMID- 16369701 TI - Predicting postoperative mortality in patients undergoing colorectal surgery. AB - Well-known and suitable instruments for surgical audit are the POSSUM and P POSSUM scoring systems. But these scores have not been well validated across the countries. The objective of the present study was to assess the predictive value of scores for colorectal surgery in France. Patients operated on for colorectal malignant or diverticular diseases, whether electively or on emergency basis, within a 4-month period were included in a prospective multicenter study conducted by the French Association for Surgery (Association Francaise de Chirurgie, AFC). The main outcome measure was postoperative in-hospital mortality. Independent factors leading to death were assessed by multivariate logistic regression analysis (AFC-index). The ratio of expected versus observed deaths was calculated, and the predictive value of the POSSUM and P-POSSUM scores were analyzed by the receiver operating characteristic (ROC) curve. A total of 1426 patients were included. The in-hospital death rate was 3.4%. Four independent preoperative factors (AFC-index) have been found: emergency surgery, loss of more than 10% of weight, neurological disease history, and age > 70 years. POSSUM had a poor predictive value; it overestimated postoperative death in all cases. P-POSSUM had a good predictive value, except for elective surgery, where it overestimated postoperative death twofold. The predictive value of the AFC-index was also good. It had the same sensitivity and specificity as the P POSSUM. POSSUM has not been validated in France in the field of colorectal surgery. P-POSSUM was as predictive as the AFC-index which is a simpler instrument based on four clinical parameters (without any mathematical formulas). PMID- 16369702 TI - Clinical characteristics and prognosis of pediatric hepatocellular carcinoma. AB - INTRODUCTION: Hepatocellular carcinoma (HCC) is a rare pediatric malignancy that is usually advanced at diagnosis, with a relatively poor prognosis. Extensive treatment, including complete tumor resection, is believed to be necessary for cure. This study was performed to analyze treatment results and to search for prognostic factors of pediatric HCC. METHODS: Between March 1982 and February 2004 a total of 16 children had been diagnosed as having HCC in our institution, and a retrospective analysis was performed. RESULTS: The median age at diagnosis was 10.5 years, and the male/female ratio was 11:5. As a predisposing condition, hepatitis B virus (HBV) infections were present in 11 (68.8%) and liver cirrhosis in 8 (50.0%). Including 1 patient with a liver transplant, 4 patients (25.0%) underwent a primary operation with complete tumor resection, and 11 (68.8%) received neoadjuvant chemotherapy because of their inoperable state at diagnosis. After neoadjuvant chemotherapy, complete tumor resection was performed in four (36.4%). Thus complete resection was undertaken in a total of eight patients (50.0%). The estimated 5-year survival rate of all patients was 27.3%. The 5-year survival rate for patients who underwent complete tumor resection was 62.5%, and for those who underwent palliative resection or no operation it was 0%. The statistically significant prognostic factors were tumor stage, presence of metastasis, and complete tumor resection. CONCLUSIONS: This study confirmed that complete tumor resection is essential for cure in pediatric patients with HCC, and neoadjuvant chemotherapy improves the tumors' resectability. PMID- 16369703 TI - Effects of peritoneal lavage with scolicidal agents on survival and adhesion formation in rats. AB - Intraoperative irrigation of the peritoneal cavity with scolicidal agents is frequently recommended when dealing with traumatic or spontaneous rupture of hydatid cysts. The present experimental study was designed to examine the influence of various scolicidal agents on adhesion formation and survival. A total of 149 rats were randomly allocated to nine groups. Peritoneal lavage through a median laparotomy was performed with the following scolicidal agents. Group 1 (0.9% saline: controls), group 2 (20% hypertonic saline), group 3 (0.04% chlorhexidine gluconate), group 4 (3% hydrogen peroxide), group 5 (0.5% silver nitrate), group 6 (1% polyvinylpyrrolidone-iodine, or PVP-I ), group 7 (5% PVP I), group 8 (0.5% cetrimide/0.05% chlorhexidine), and group 9 (10% PVP-I). The surviving animals were sacrificed on postoperative day 15. Adhesion formation was macroscopically graded by the Nair criteria. The severity of adhesion formation was evaluated microscopically using the fibrosing scoring criteria and the strain test. Group 9 (10% PVP-I) was excluded from the adhesion evaluation because all of the rats died in this group. The mortality rate was significantly higher in groups 5 and 7 than in groups 1, 2, 3, 4, 6, and 8. Adhesion scores were significantly lower in groups 1, 2, 3, and 4 than in groups 5, 6, 7, and 8. The lowest adhesion score was found in group 3 and the highest in the group 7. These results indicate that 0.04% chlorhexidine gluconate, the most potent scolicidal agent in vitro and in vivo, was associated with the lowest adhesion formation and mortality among various scolicidal agents in this experimental study. PMID- 16369704 TI - Complications following D3 gastrectomy: post hoc analysis of a randomized trial. AB - INTRODUCTION: A single institutional surgical trial for gastric cancer had demonstrated increased morbidity but not mortality. This report analyzes risk factors affecting morbidity. METHODS: Risk factors for morbidity in 221 patients treated with curative intent were evaluated in a prospective randomized trial comparing D1 and D3 surgery for curable gastric cancer. RESULTS: The surgeon's experience after 25 nodal dissections had no influence on surgical or overall complications, nor did the patients' co-morbidity (e.g., respiratory system disease, cardiac disease, diabetes mellitus). Distal pancreatectomy negatively affected surgical morbidity [relative risk (RR) 6.21, 95% confidence interval (CI) 1.869-20.626] and overall morbidity (RR 5.50, 95% CI 1.671-18.082). All of the patients with a distal pancreatectomy underwent concomitant splenectomy. Multivariate analysis found splenectomy and nodal dissection to be the only two independent risk factors adversely affecting operative morbidity. The RR of splenectomy for surgical complications was 4.19 (95% CI 1.327-13.208), and for overall complications it was 3.88 (95% CI 1.259-11.973). The RR of nodal dissection for surgical complications was 2.51 (95% CI 1.336-4.730), and for overall complications it was 1.93 (95% CI 1.149-3.255). CONCLUSIONS: Splenectomy (with or without pancreatectomy) and nodal dissection are risk factors for operative morbidity but not mortality. PMID- 16369705 TI - Prognostic factors for sporadic medullary thyroid carcinoma. AB - INTRODUCTION: Medullary thyroid carcinoma (MTC) originates from the thyroid parafollicular cells and accounts for 3% to 10% of all thyroid malignancies. Approximately 84% of cases are sporadic. The aim of this study was to evaluate the outcomes of treatment for sporadic medullary thyroid carcinoma (SMTC) and define the prognostic factors for overall survival. METHODS: The records of 32 SMTC patients treated at Ankara Oncology Education and Research Hospital between September 1993 and April 2003 were retrospectively evaluated. The effects of age, gender, tumor localization, extent of the primary surgical resection, tumor size, capsule invasion, lymph node metastasis, extranodal extension, tumor stage, local recurrence, and distant metastasis on the overall survival rate were evaluated by univariate and multivariate analyses. RESULTS: There were 32 patients (19 females, 13 males) with a median age of 45 years (21-76 years). Altogether, 22 patients had undergone complete resection and 10 patients incomplete resection. The median follow-up was 48 months (9-111 months), and the overall 5-year survival rate was 51%. Based on the univariate analysis, the extent of primary surgical resection, pathologic tumor size, capsule invasion, lymph node invasion, extranodal extension, tumor stage, local recurrence, and distant metastasis were factors that significantly affected survival. In the multivariate analysis, however, only the extent of the primary surgical resection, capsule invasion, and distant metastasis were found to be statistically significant factors. CONCLUSIONS: The extent of the primary surgical resection significantly influences the survival of patients with SMTC. Capsule invasion and distant metastasis were additional factors affecting the prognosis. PMID- 16369706 TI - Role of interval appendectomy in the management of complicated appendicitis in children. AB - INTRODUCTION: The aim of this study was to ascertain the optimal treatment for children with complicated appendicitis. We reviewed an inception cohort of children with documented complicated appendicitis to develop criteria for interval appendectomy. We compared the outcomes of two treatments: immediate operation and interval appendectomy. METHODS: Children with complicated appendicitis were separated into two groups. Group 1 patients had had symptoms of complicated appendicitis for less than 72 hours or appeared toxic. Group 2 patients had had symptoms of complicated appendicitis for longer than 72 hours and did not appear toxic. Group 1 underwent immediate operation treated by criteria previously published. Group 2 patients were treated in hospital with triple antibiotics until they were afebrile, had normal white blood cell counts, tolerated an oral diet, and had adequate pain control. They were discharged on oral metronidazole or metronidazole plus Bactrim for 6 weeks and then underwent interval appendectomy. RESULTS: A total of 86 children had complicated appendicitis; 59 were operated on immediately, and 27 underwent an interval appendectomy. Complications included one wound infection and two intraabdominal abscesses (all in group 1). There was one death (group 1). The length of stay for the immediate operation group was 4.9 +/- 1.7 days; the initial-admission length of stay for the interval appendectomy group was 4.1 +/- 1.0 days with a subsequent postoperative stay of 0.9 +/- 0.8 days. One patient in the interval appendectomy group was treated off protocol. CONCLUSIONS: Treating selected children with interval appendectomy led to a decrease in complications and a shorter length of stay in this limited population. Interval appendectomy is a safe, cost-effective, useful adjunct treatment for children with complicated appendicitis. PMID- 16369707 TI - Surgical management of small gastrointestinal stromal tumors of the stomach. AB - Small gastrointestinal stromal tumors (GISTs) (< 3 cm) occasionally are found in the stomach during endoscopy. There is no consensus about the surgical management of these small tumors, although this clinical issue is crucial because some of the tumors show unexpected malignant behavior. In this study, we evaluated the clinical management of patients with gastric GISTs who underwent surgical resection. Altogether, 31 patients with gastric GISTs were examined retrospectively. Surgical resection was fundamentally indicated for the patients with gastric GISTs suspected to be malignant by endoscopy or endoscopic ultrasonography (EUS). The malignant grade of the GISTs was evaluated by the mitotic rate, tumor size, and MIB-1 index. EUS was useful for differentiating benign from malignant GISTs; but by limiting the study to patients with small tumors (< 3 cm), the diagnostic value of EUS was not satisfactory for defining the surgical indication. Tumors that were < 50 mm were successfully treated by laparoscopic surgery. Of the 31 patients, 4 had a relapse of the disease, and 1 of those 4 patients had a small tumor (30 mm). All of the recurrences were classified in the high risk category. Surgery is indicated for gastric GISTs that are > or = 20 mm or are suspected to be malignant based on EUS findings. Laparoscopic resection is feasible and is recommended as the treatment of choice for patients with tumors < 50 mm. Risk assessment can be most useful for predicting recurrence. PMID- 16369708 TI - Successful repair of iatrogenic rectourinary fistulas using the posterior sagittal transrectal approach (York-Mason): 15-year experience. AB - INTRODUCTION: Rectourinary fistulas (RUFs) represent a challenging clinical problem. Most RUFs are secondary to lower urinary or intestinal tract surgery. Several surgical approaches have been proposed. The aim of this study was to review a 15-year experience using the York-Mason posterior sagittal transrectal approach to iatrogenic RUFs. METHODS: Seven patients with RUFs secondary to urologic surgery were operated on with the York-Mason technique at the Department of Urology, University of Padova, Italy between 1988 and 2003. The patients' data have been collected and analyzed retrospectively. RESULTS: All the patients were treated successfully (100%). In one patient with Crohn's disease the fistula recurred 11 years after the first surgery. One patient died for metastasis of prostate cancer 1 year after surgical repair of the RUF. A temporary colostomy was performed in five patients; the colostomies were subsequently closed, and the patients regained complete fecal continence with no postoperative anal strictures. The colostomy remained in place in one patient with Crohn's disease and in another with ulcerative rectocolitis. CONCLUSIONS: The posterior sagittal transrectal approach provided easy access and identification of RUFs and good surgical exposure, with no subsequent strictures or fecal incontinence. Our data show that the York-Mason technique alone is a highly effective option for treating an iatrogenic postoperative RUF. PMID- 16369709 TI - Survival of young patients after gastrectomy for gastric cancer. AB - It has been suggested that gastric cancer has a worse prognosis in young patients, but the data are controversial. The aim of this study was to compare the 5-year survivals after gastrectomy for gastric cancer in two groups of patients (those < or = 45 years of age and those (> 45 years) and to determine some of the prognostic factors. The 5-year survival was significantly better for patients < or = 45 years of age. Survival was also better for young patients with a curative resection and also for those with lymph node metastases. However, survival was not significantly different for the two groups when the resection was not curative and when the lymph nodes were not involved. Survival was no different for the two groups when compared at each stage, although a multivariate analysis showed that age > 45 years, moderate or poor degree of differentiation of the tumor, advanced tumors, the presence of lymph node involvement, and a noncurative resection were independent negative prognostic factors. Long-term survival after gastrectomy for gastric cancer depends on the stage of the disease; the age of the patient is not a decisive factor. PMID- 16369710 TI - Technetium-99m-MIBI SPECT/CT in primary hyperparathyroidism. AB - The novel trend toward focused parathyroidectomy requires precise preoperative localization of the parathyroid adenoma in patients with primary hyperparathyroidism (PHPT). The present study evaluated the impact of hybrid single photon emission computed tomography/computed tomography (SPECT/CT), using 99mTc-sestamibi (MIBI), on the surgical management of these patients. In a retrospective study of 36 patients with PHPT, SPECT/CT was undertaken when planar 99mTc-MIBI scintigraphy was negative or when an ill-defined focus in the neck or an ectopic site on planar views was visualized. Imaging data were compared with intraoperative findings, and the incremental value of SPECT/CT to lesion localization and surgical procedure was assessed. Three patients with both negative planar and SPECT/CT studies subsequently underwent bilateral neck exploration, with multiglandular hyperplasia diagnosed in two patients and a parathyroid adenoma in one. Of 33 patients with a positive MIBI study, parathyroid adenoma was confined to the neck in 23 patients and to the lower neck mediastinum in 10. SPECT/CT facilitated the surgical exploration of all 10 ectopic parathyroid adenomas and 4 of 23 cervical parathyroid adenomas, the latter four either at reexploration or in patients with nonvisualization of the thyroid after thyroidectomy. SPECT/CT contributed to the localization of parathyroid adenomas in patients with PHPT and to planning the surgical exploration in 14 of 36 (39%) patients, predominantly those with ectopic parathyroid adenomas or who had distorted neck anatomy. PMID- 16369711 TI - Early presumptive therapy with fluconazole for occult Candida infection after gastrointestinal surgery. AB - The objective of this retrospective comparative study was to improve the outcome of patients with suspected occult Candida infection after gastrointestinal surgery by early presumptive therapy. It was conducted in the National Cheng Kung University Hospital in Taiwan. A total of 36 patients with prolonged ileus with fever after gastrointestinal tract surgery between January 1995 and December 2002 were examined for two time periods: those treated before and those treated after January 1999. One set of patients did not receive early presumptive therapy (EPT) until Candida infection was confirmed, and they were designated EPT(-). Another group of patients with suspected occult Candida infection received EPT and were designated EPT(+). Fluconazole, 400 mg/day, was given as EPT. Urine, wound, intraperitoneal drainage, and blood specimens were obtained from patients for fungus culture before starting treatment and weekly until symptoms subsided. The primary endpoints were the frequency of candidiasis and the persistence of candidemia; the secondary endpoint was the efficiency of EPT in the clinical outcome. There was no difference in Candida peritonitis, wound colonization, or urine colonization in the two treatment groups. Candida albicans accounted for 87.5%% of the isolated Candida species: 84.6%% in the EPT(+) group and 89.5%% in the EPT(-) group. In the EPT(+) group, the positive blood culture rate was 66.7%%. The fever subsided rapidly in 17 patients (94%%), the hospital stay and intensive care unit stay were shorter, and the mortality decreased significantly: 11%% vs. 78%%, P < 0.001. Persistent gastrointestinal ileus was the main cause of breakthrough candidemia. We concluded that EPT with fluconazole improves the prognosis of patients with occult Candida infection after gastrointestinal surgery. Surgical intervention was required in patients with breakthrough candidemia. PMID- 16369712 TI - The outcome of cervical exploration for asymptomatic and symptomatic patients with primary hyperparathyroidism. AB - This study examined the success and safety of cervical exploration in patients with primary hyperparathyroidism (HPT). The presentation, pathologic findings, and outcome of patients with asymptomatic primary HPT were compared with those with symptomatic disease. Records of patients undergoing cervical exploration for primary HPT from January 1993 until December 31, 2003, were reviewed. Information collected consisted of preoperative symptoms, calcium and parathormone (PTH) levels, imaging studies, operative findings, pathology, and outcome of the patients. The groups with asymptomatic and symptomatic primary HPT were compared. In all, 139 patients were studied; 31 (22.3%) were asymptomatic (group I), and 108 (77.7%) had symptoms (group II). The two groups were also comparable regarding mean age, sex, and the yield of the imaging studies. The mean preoperative serum calcium level was comparable in the two groups (11.1 mg/dl versus 11.3 mg/dl). However, PTH levels were significantly lower in group I than in group II (142 pg/dl versus 283 pg/dl, P = 0.01). The weight of the adenoma was also significantly less in group I than in group II (1082 mg versus 1679 mg P = 0.079). The outcome of the surgical exploration was comparable in the two groups with an immediate success rate close to 98% and a long-term success rate of 95.4%. Cervical exploration and parathyroidectomy in patients with primary HPT is a safe procedure with a high success rate and favorable outcome. PMID- 16369713 TI - Changing treatment strategy for gastrinoma in patients with Zollinger-Ellison syndrome. AB - We overviewed the recent development of curative surgery for gastrinoma that has been rapidly improved since the development of new localization techniques, especially the selective arterial secretagogue injection test (SASI test) and somatostatin receptor scintigraphy (SRS). A number of new pathological findings of gastrinomas in patients with Zollinger-Ellison syndrome have been accumulated in accordance with the increase of curative resection of gastrinomas, and these new findings also have contributed to the progress of the treatment strategy for grastrinomas. PMID- 16369714 TI - Incidence and impact of risk factors in critically ill trauma patients. AB - There is a paucity of data describing the incidence of pre-existing diseases or risk factors and their effects in trauma patients. We conducted a prospective study to determine the incidence of such factors in critically ill trauma patients and to evaluate their impact on outcome. The study, performed over a 2 year period, examined the hospital course of all trauma patients admitted to the ICU. Multiple risk factors were evaluated and analyzed via multivariate regression analysis. Outcome was evaluated by infection rate, hospital length of stay, ventilator days, and mortality matched for age and Injury Severity Score (ISS). A total of 1172 patients (73% blunt injury) were enrolled over the study period. Of these, 873 (74.5%) were male. The mean age was 42.5 years with an ISS of 19.8. Tobacco use (24%) was the most common risk factor identified, followed by hypertension (HTN, 17%), coronary artery disease (9%), chronic obstructive pulmonary disease (COPD)/reactive airway disease (4%), non-insulin-dependent diabetes (NIDDM) (4%), insulin-dependent diabetes (IDDM) (3.2%), cancer (3%), liver disease (2%), and HIV/AIDS (1.4%). Of these risk factors, IDDM was found to be an independent risk factor for infection (0.004) and ventilator days (0.047), increasing age was found to be an independent risk factor for hospital length of stay (0.023) and mortality (<0.001), and HTN was found to be an independent risk factor for increased ventilator days (0.04). In addition, COPD/reactive airway disease was found to be an independent predictor of ventilator days, infection, and ICU days (P < 0.05). Thus, increased age, IDDM, COPD, and HTN are most predictive of outcome in critically ill trauma patients. With our aging population it is becoming increasingly important to identify pre-existing risk factors on admission in order to minimize their effects on outcome. PMID- 16369715 TI - Significance of extensive surgery including resection of the pancreas head for the treatment of gallbladder cancer--from the perspective of mode of lymph node involvement and surgical outcome. AB - The present study aimed to clarify the efficacy of extensive surgery, including pancreas head resection, for more complete lymphadenectomy in the treatment of gallbladder carcinoma. The study involved retrospective analyses of 65 consecutive patients with gallbladder carcinoma who underwent surgical resection between 1982 and 2003. Of these 65 patients, 41.5% displayed node-positive disease and among them 23.1% had positive para-aortic nodes. Of six node-positive 5-year survivors, five underwent pancreatoduodenectomy combined with S4aS5 hepatic subsegmentectomy. The 5-year survival rates were 76.2% for pN0, 30.0% for pN1, 45.8% for pN2, and 0% for pM1[lymph], respectively. Significant differences existed in survival rates. Postoperative recurrence was observed in 24.1% (13/54) of patients who underwent R0 resection. Of the four patients who displayed lymph node recurrence, two had pericholedocal and/or posterior pancreatoduodenal lymph node metastasis at the time of surgery and underwent pancreas-preserving regional lymphadenectomy. These results suggest that extensive resection, including resection of the pancreatic head, is effective in selected patients with up to pN2 lymph node metastasis, as long as complete removal of the cancer can be achieved. Pancreatoduodenectomy combined with S4aS5 hepatic subsegmentectomy should be considered when lymph node metastasis is obvious and the patient is in good condition. PMID- 16369716 TI - Anticipated detection of imminent surgeon-patient barrier breaches. A prospective randomized controlled trial using an indicator underglove system. AB - The double gloving indicator underglove system (IUS) is based on a colored detection of the outer glove perforation. Our objective was to determine the IUS efficiency to detect outer glove perforations and to reduce the risks of blood and body fluids exposure, warning the surgeon before the breach of the surgeon patient barrier (SPB). A series of 100 visceral surgical procedures were randomly assigned to either double (IUS) or single gloving. The noticed glove perforations (using the water test method) and the IUS efficiency were analyzed in 99 procedures. In 49 single-gloving procedures, 19 perforations were noticed: one was immediately perceived (perceived accidental exposure, PAE); 3 were discovered as the gloves were being removed, and 15 were undetected before the water test (unperceived prolonged contact, UPC). In 50 double-gloving procedures (IUS), 16 perforations were noticed, all of them involving only the outer glove: the IUS allowed immediate detection of 3 perforations without any blood exposure; 13 other perforations went undetected but without any UPC. In conjunction with the protective quality of double gloving, the IUS allows detection of significant breaches of the outer glove before the breach of the SPB. PMID- 16369717 TI - Results of preoperative lymphoscintigraphy for breast cancer are predictive of identification of axillary sentinel lymph nodes. AB - The aim of this study was to identify the variables associated with successful peroperative sentinel lymph node (SLN) localization. We studied 201 patients with T1, T2, N0 invasive breast cancer who underwent a SLN procedure from 1999 to 2003. Of these 201 patients, 55 underwent peritumoral and 146 underwent periareolar radioisotope injection before the blue dye injection. All patients were operated on by breast conservative surgery and axillary dissection after SLN biopsy. Age, weight, menopausal status, previous biopsy, localization of the tumor, results of lymphoscintigraphy, site of radiotracer injection, tumor size, tumor grade, experience of surgeons, and the number of invaded axillary nodes were analyzed to determine whether they had any significant correlation with successful identification of SLN. Variables found to have a statistically significant influence on the SLN identification rate and on preoperative lymphoscintigraphy identification were introduced into a univariate and multivariate logistic regression model. In multivariate analysis, successful lymphoscintigraphy (P < 0.0001) and the absence of metastatic axillary nodes (P < 0.005) were associated with successful identification of SLNs. The peritumoral injection of radiotracer (P < 0.001), patient age > 60 years (P < 0.003), and localization of the tumor in the upper outer quadrant (P < 0.004) were associated with failure of lymphoscintigraphic visualization of SLN. The technique of SLN detection thus appears to be better for patients with low risk of invaded axillary lymph nodes. PMID- 16369718 TI - Is there a role for palliative gastrectomy in patients with stage IV gastric cancer? AB - Patients with metastatic gastric cancer are currently not considered operative candidates and are most often offered systemic therapy. Palliative resection of the primary tumor has been considered irrelevant to the outcome and has been recommended only for palliation of symptoms. We have examined the role of palliative gastrectomy and its impact on survival in patients with stage IV gastric cancer at initial diagnosis between 1990 and 2000. A total of 105 patients with stage IV disease were identified during this period; 81 of them (77.1%) had no resection, and 24 (22.9%) underwent palliative gastric resection. Mean survival in those without resection who received chemotherapy (with or without radiation) treatment was 5.9 months (95% confidence interval 4.2-7.6). For those with resection and adjuvant therapy, mean survival time was 16.3 months (95% confidence interval 4.3-28.8 months). Kaplan-Meier survival analysis showed significantly better survival in those with resection and adjuvant therapy (log rank test, P = 0.01). Mortality and morbidity rates associated with palliative resection were 8.7% and 33.3%, respectively, which did not differ statistically from the 3.7% and 25.3% in patients who underwent curative gastrectomy during same period of time. However, the length of hospitalization (22 versus 16 days) was significantly higher compared with those without stage IV disease. These data suggest that palliative resection combined with adjuvant therapy may improve survival in a selected group of patients with stage IV gastric cancer. Palliative gastrectomy plus systemic therapy should be compared with systemic therapy alone in a randomized trial. PMID- 16369719 TI - Treatment of mammary duct fistula by fistulectomy and saucerization. AB - This study was designed to assess the efficacy and long-term outcome of fistulectomy and saucerization for treatment of mammary duct fistulae. Mammary fistula is a chronic condition that represents the final step in what has been termed "mammary duct associated inflammatory disease sequence." The treatment is primarily surgical and may include healing by secondary intention or primary closure with or without antibiotics. Reported series are small and often include variable surgical strategies applied without consistency. A consecutive series of 53 patients who had 59 mammary duct fistulae were treated by fistulectomy with saucerization. The median age was 32 years. Wounds were allowed to heal by secondary intention and antibiotics were not used. We reviewed the case records to establish the incidence of recurrent fistula and the time to complete healing. The long-term cosmetic outcome was determined by a postal survey. After a median follow-up of 6 years there had been no relapse in 92%. There was significant delay in healing in six cases (range: 10 to 30 weeks). Thirty-eight patients (83%) gave a definite history of regularly smoking between 10 and 20 cigarettes a day. Two thirds of the patients were either pleased or satisfied with the final cosmetic result of the surgery, but more than 90% said that it left them with some distortion of the nipple. Fistulectomy and saucerization achieves long-term cure in the majority of patients with mammary duct fistula, but it results in some degree of distortion of the nipple. The strong relationship between smoking and the occurrence of mammary duct fistulae is again demonstrated. PMID- 16369721 TI - Clinical significance of metastasis to the central compartment from papillary microcarcinoma of the thyroid. AB - We previously demonstrated that for papillary microcarcinoma (PMC), (1) patients with lateral lymph node metastasis detected on preoperative ultrasonography (US) are more likely to develop recurrence, and (2) dissection of the lateral compartment does not improve the prognosis of patients without US-detectable lateral metastasis. In this study, we focused on metastasis from PMC to the central compartment. We investigated the clinical significance of lymph nodes in the central compartment using the same series of 600 patients as used in the previous study. Ultrasound-diagnosed central node metastasis could be found in 30 patients (20.0%). Although the specificity of US for detecting central node metastasis was 99.1%, sensitivity was only 10.9%. Neither US-diagnosed nor pathologically confirmed central node metastasis affected the disease-free survival (DFS) rate of PMC patients. Furthermore, in cases where PMC was located only in one lobe, central node dissection in the contralateral lobe did not improve the DFS rate. These findings suggest that, for PMC, (1) US is an insensitive technique for detecting the central node metastasis, (2) the presence of central node metastasis is unrelated to DFS of patients, and (3) when performing lobectomy for PMC involving only one lobe, dissection of the central compartment in the contralateral lobe is optional. PMID- 16369723 TI - Surgical treatment of papillary thyroid carcinoma. PMID- 16369725 TI - Reduction of sampling bias of odds ratios for vertebral fractures using propensity scores. AB - INTRODUCTION: Assessment of the predictive power of a newly introduced diagnostic technique with regard to fracture risk is frequently limited by the enormous costs and long time periods required for prospective studies. A preliminary estimate of predictive power usually relies on cross-sectional case-control studies in which bone measurements of normal and fractured subjects are compared. The measured discriminatory power is taken as an estimate of predictive power. Because of possible sample selection bias, study participants may have different bone mineral density (BMD) values, and fractured patients may have fractures of different severity levels. The same diagnostic techniques for the measured discriminatory power, expressed as odds ratios, will differ among studies with different patient and control populations. METHODS: In this paper, we propose a weighted logistic regression approach to adjust the odds ratio in order to reduce the effect of sampling bias. The weight is derived from age, deformity severity, BMD, and the interactions of these, using the propensity score theory and reference population data. RESULTS: Simulation examples using data from the Osteoporosis and Ultrasound Study (OPUS) demonstrate that such a procedure can effectively reduce the estimation bias of odds ratios introduced by sampling differences, such as for dual x-ray absorptiometry (DXA) scans of the spine and hip as well as various quantitative ultrasound techniques. The derived estimated odds ratios are substantially less biased, and the corresponding 95% confidence intervals contain the true odds ratios from the population data. CONCLUSIONS: We conclude that a statistical correction procedure based on propensity scores and weighted logistic regression can effectively reduce the effect of sampling bias on the odds ratios calculated from cross-sectional case-control studies. For a new diagnostic technique, hip BMD and deformity severity information are necessary and likely sufficient to derive the propensity scores required to adjust the measured standardized odds ratios. PMID- 16369728 TI - Effect of sternotomy and extracorporeal circulation on pulmonary neutrophil kinetics in pigs. AB - Pulmonary margination of neutrophils may contribute to lung damage after extracorporeal circulation for cardiac surgery. We evaluated single-pass pulmonary neutrophil kinetics using the multiple indicator-dilution technique in control pigs (n = 10), after sternotomy alone (sterno, n = 10) or after 30 min of observation following a period of 90 min extracorporeal circulation (n = 7). Blood neutrophils increased in the control and sterno groups (p < 0.05) but remained unchanged in the extracorporeal circulation group. The transfer coefficient for neutrophil margination from the circulating to the lung marginated pool (k(c-m)) and pulmonary neutrophil clearance (Cl(c-m)) were similar between the three groups. There was an inverse correlation between k(c-m) and the degree of lung tissue perfusion evaluated from the tracer-accessible extravascular lung water (r = -0.54, p < 0.01). There was no arterio-venous gradient of neutrophils in any of the groups, suggesting a dynamic equilibrium of the margination/demargination processes. We conclude that extracorporeal circulation does not significantly modify single pass pulmonary neutrophil kinetics 30 min after reperfusion. The rate of neutrophil margination to the tracer-accessible lung tissue suggests that lung tissue de-recruitment is associated with increased neutrophil margination. PMID- 16369727 TI - Vascular endothelial growth factor delays onset of failure in pressure-overload hypertrophy through matrix metalloproteinase activation and angiogenesis. AB - OBJECTIVE: Pressure-overload hypertrophy is associated with decreased capillary density in myocardium resulting in impaired substrate delivery. Treatment of hypertrophied hearts with vascular endothelial growth factor (VEGF) induces angiogenesis. Since angiogenesis is associated with extracellular matrix degradation, we sought to determine whether VEGF induced angiogenesis in hypertrophy required matrix metalloproteinases (MMP) activation. METHODS: Newborn rabbits underwent aortic banding. Progression of hypertrophy (mass-to-volume (M/V) ratio) and mid-wall contractility index was monitored by echocardiography. At 4 and 6 weeks, VEGF (2 microg/kg), vehicle or VEGF combined with GM6001 (5 mg/kg), a MMP inhibitor, was administered intrapericardially. CD-31 (indicator of angiogenesis), MMP-2, MT1-MMP and TIMPs (endogenous MMP inhibitors) expression were measured by immunoblotting. MMP-2 activity was determined by gelatin zymography. RESULTS: Untreated hypertrophied hearts progressed to ventricular dilatation at 7 wks (M/V ratio: 0.75 +/- 0.07), but compensatory hypertrophy was maintained with VEGF (0.91 +/- 0.07; p < 0.05). LV contractility declined in untreated hearts from -0.41 +/- 0.9 (5 wks) to -0.73 +/- 0.5 (7 wks; p < 0.05) but remained normal with VEGF (+1.61 +/- 0.6 vs. +0.47 +/- 0.2). MMP-2 expression and activity were significantly elevated in VEGF treated hypertrophied hearts (p < 0.05) and were blocked by concomitant administration of GM6001. VEGF induced neovascularization was inhibited by addition of GM6001. MT1-MMP showed a trend to higher levels in VEGF treated hearts. TIMPs were unchanged in all three groups. CONCLUSIONS: Exogenous VEGF and resultant MMP-2 activation leads to increased capillary formation in severe hypertrophy, preventing progression to ventricular dilation and dysfunction. VEGF and the associated MMP-2 activation play an important and potentially therapeutic role in vascular remodeling of hypertrophied hearts. PMID- 16369729 TI - Prevalence of unidirectional Na+-dependent adenosine transport and altered potential for adenosine generation in diabetic cardiac myocytes. AB - Adenosine is an important physiological regulator of the cardiovascular system. The goal of our study was to assess the expression level of nucleoside transporters (NT) in diabetic rat cardiomyocytes and to examine the activities of adenosine metabolizing enzymes. Isolated rat cardiomyocytes displayed the presence of detectable amounts of mRNA for ENT1, ENT2, CNT1, and CNT2. Overall adenosine (10 microM) transport in cardiomyocytes isolated from normal rat was 36 pmol/mg/min. The expression level of equilibrative transporters (ENT1, ENT2) decreased and of concentrative transporters (CNT1, CNT2) increased in myocytes isolated from diabetic rat. Consequently, overall adenosine transport decreased by 30%, whereas Na(+)-dependent adenosine uptake increased 2-fold, and equilibrative transport decreased by 60%. The activity ratio of AMP deaminase/5' nucleotidase in cytosol of normal cardiomyocytes was 11 and increased to 15 in diabetic cells. The activity of ecto-5'-nucleotidase increased 2-fold in diabetic cells resulting in a rise of the activity ratio of ecto-5'-nucleotidase/adenosine deaminase from 28 to 56.These results indicate that in rat cardiomyocytes diabetes alters activities of adenosine metabolizing enzymes in such a way that conversion of AMP to IMP is favored in the cytosolic compartment, whereas the capability to produce adenosine extracellularly is increased. This is accompanied by an increased unidirectional Na(+)-dependent uptake of adenosine and significantly reduced bidirectional adenosine transport. PMID- 16369730 TI - Reduced serine-16 and threonine-17 phospholamban phosphorylation in stunning of conscious dogs: no evidence for any involvement of protein kinase A or protein phosphatases. AB - OBJECTIVE: Cardiac stunning is the consequence of a brief cardiac ischemia. The underlying mechanism is not completely understood. METHODS: Here we induced cardiac transient ischemia in conscious instrumented dogs by means of an occluder in the left anterior descending coronary artery (LAD). Contractile performance, monitored by ultrasound crystals, was reduced during and after ischemia in the LAD area. For control in the same animals cardiac performance was measured in the area of left circumflex coronary artery (Ramus circumflexus, RCx). In the RCx area, no decline in contractility was noted. Tissue was obtained from stunned LAD area and from control areas (RCx). RESULTS: Phospholamban phosphorylation on both serine-16 and threonine-17 was reduced in LAD areas compared to RCx areas. Reduced phosphorylation of PLB is known to inhibit cardiac contractility. While phosphorylation of PLB was reduced, the activity of the appropriate protein phosphatases and protein kinases was not different between tissue obtained from LAD or RCx areas. CONCLUSION: Reduced formation of cAMP might underlie the contractile dysfunction in myocardial stunning. PMID- 16369731 TI - Bovine lactoferrin is effective to suppress Helicobacter pylori colonization in the human stomach: a randomized, double-blind, placebo-controlled study. AB - Bovine lactoferrin (bLF) has antibacterial activity against Helicobacter pylori in vitro and is effective to suppress bacterial colonization in mice. The aim of our study was to evaluate the efficacy of orally administered bLF on H. pylori colonization in humans by a randomized, double-blind, placebo-controlled study. Fifty-nine healthy subjects positive for H. pylori infection were recruited. Subjects were randomized into two groups. The bLF group received bLF tablets at a dosage of 200 mg b.i.d. for a period of 12 weeks, and the control group received placebo tablets without bLF. The (13)C-urea breath test (UBT) was performed before, during, and at the end of administration, and again 4 weeks after administration. Positive response was defined as more than 50% decrease of the UBT value at the end of administration. Positive response was observed in 10 of 31 bLF-treated subjects (32.3%) and 1 of 28 control subjects (3.6%), indicating that the rate of positive response in the bLF group was significantly higher than that in the control group (bLF vs. control, P < 0.01). These results suggested that bLF administration is effective to suppress H. pylori colonization. PMID- 16369732 TI - Incidence of sexually transmitted infections in asymptomatic healthy young Japanese men. AB - The aim of this study was to estimate the incidence rates of asymptomatic sexually transmitted infections (STI) in young men in Sapporo, Japan. A survey of 204 healthy male volunteers was conducted. First-voided urine specimens were used for detection of Chlamydia trachomatis and Neisseria gonorrhoeae with polymerase chain reaction assay, and cotton swab wiping of the external genitalia was used for detection of human papillomavirus (HPV) with the Hybrid Capture method. The incidence rates were 3.4% for C. trachomatis and 5.9% for HPV. No N. gonorrhoeae was detected. In sexually active men who had regular sexual intercourse, the detection rate of C. trachomatis was 4.7% and that of HPV was 8.0%. Our study showed that the incidence rates of asymptomatic STIs were not negligible, and that we should provide screening programs for young men with a high risk for such unrecognized infections and study their natural histories. PMID- 16369733 TI - Clinical effect of gatifloxacin, 200 mg daily, on bacterial community-acquired pneumonia in the elderly. AB - The clinical effect of gatifloxacin (GFLX) at 200 mg daily (half of common dosage) on bacterial community-acquired pneumonia in the elderly was examined. Subjects were 30 patients with bacterial community-acquired pneumonia aged 65 years or more (mild, 21 patients; moderate, 9 patients), and GFLX at 100 mg per dose was administered twice daily for 4-14 days. The results included clinical effectiveness in 21 of 21 mildly affected patients (efficacy rate, 100%), in 8 of 9 moderately affected patients (efficacy rate, 88.9%), and 29 of a total of 30 patients (efficacy rate, 96.7%). With regard to bacteriological effect, 28 of 29 strains were eradicated (eradication rate, 96.6%). Abnormal laboratory findings included mild elevations in GPT, GOT, and ALP in only 1 patient. Based on these findings, we concluded that administration of GFLX at 200 mg daily is recommended for bacterial community-acquired pneumonia in the elderly. PMID- 16369734 TI - Current status of diagnosis and treatment of invasive fungal infections in Japan: the influence of the new Japanese guidelines. AB - One year after the release of the Japanese "Guidelines for the Diagnosis and Treatment of Deep-Seated Mycosis" we conducted a survey of clinicians to determine the extent to which the new guidelines had penetrated clinical practice, and compared these results with those of a previous survey. The responses to the current survey regarding the diagnosis and treatment had changed from those of the previous survey to reflect the new recommendations, showing that the guidelines have had an effect on clinical practice. However, the current survey highlights a need to provide more practical information in the guidelines for use in the clinical settings. In particular, physicians expect the guidelines to proactively provide more information about the features of both current and new drugs. In addition, an effective drug against the genus Aspergillus is eagerly awaited. However, because it is difficult to differentiate among filamentous fungi, there is a need for a drug with broad-spectrum coverage against filamentous fungi. Investigation of combination therapy consequently becomes necessary. Definitive diagnoses of invasive fungal infection are too scarce at the national level. The cooperation of clinicians for organizing more definitive diagnoses would be appreciated when the guidelines are revised. PMID- 16369735 TI - Screening of antibiotics resistance to Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii by an advanced expert system. AB - The VITEK2 advanced expert system (AES) gives information about the antibiotics resistance mechanisms based on the biological validation derived from the VITEK2 susceptibility result. In this study, we investigated whether or not this system correctly categorized the beta-lactamase resistance mechanism data derived from the VITEK2 susceptibility result using the testing card, AST-N025, with Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii. We used 131 strains, and their phenotypes were determined according to the biological and genetic screening. The AES analysis result matched the phenotype testing in 120 (91.6%) of the 131 strains. Incorrect findings were found in six strains, including three strains of Serratia marcescens. The resistance mechanism could not be determined in five strains, including three strains of Providencia rettgeri. The analysis of those phenotypes agreed in 34 (97.1%) among 35 strains with extended spectrum beta-lactamase (ESBL), and in 27 (96.4%) among 28 strains with high-level cephalosporinase. The agreement ratio in the phenotype was very high as we expected. The incorrect and nondeterminable samples were strains with relatively high cephalosporinase that has variation of outer membrane protein. The AES was able to detect the phenotype for carbapenemase. The AES is a clinically useful system that allows taking prompt measures to treat patients because it can provide information about the resistance mechanism in less than half a day after starting the analysis. PMID- 16369736 TI - Infectious bulla of the lung caused by Mycobacterium intracellulare. AB - A healthy 24-year-old man who was admitted to our hospital was suspected of having an infectious bulla of the lung because an intrabullous effusion was seen on a chest radiograph. A bulla of the right lung had been found 2 years earlier during a periodic health examination. On a chest radiograph, a bullous lesion with a niveau-like shadow (14 x 7 cm) was noted in the right lower lobe of the lung. On puncture aspiration, the bulla fluid was found to be exudative and contained mostly lymphocytes. A smear examination for acid-fast bacilli was positive, as was polymerase chain reaction (PCR) examination for Mycobacterium intracellulare. Therefore, a diagnosis of infectious lung bulla caused by M. intracellulare was made. A right lower lobectomy was performed. On macroscopic examination of the resected lesion, the area surrounding the bulla was whitish in color, and microscopic examination of this area showed a caseating epitheloid granuloma with acid-fast bacilli. We report this case because there have been no previous reports in the literature of infectious lung bulla caused by M. intracellulare although there have been several reports dealing with lung bulla caused by Mycobacterium tuberculosis. PMID- 16369737 TI - Abdominal aortic mycotic aneurysm, psoas abscess, and aorto-bisiliac graft infection due to Salmonella typhimurium. AB - Infections due to nontyphoidal Salmonella are common and their incidence has been increasing in the last few years. Here, we describe a patient with a rupture of abdominal aortic aneurysm associated with a psoas abscess due to Salmonella typhimurium. Early diagnosis, prompt surgical intervention, and active and prolonged antibiotic therapy are the gold standard for the management of this severe clinical situation. PMID- 16369738 TI - Clinical relevance of nucleic acid amplification test for patients with urinary tuberculosis during antituberculosis treatment. AB - The nucleic acid amplification test (NAAT) has been valuable in the diagnosis of urinary tuberculosis; however, no studies have attempted to determine the significance of NAAT post treatment. We encountered three patients with urinary tuberculosis who underwent sequential NAAT during antituberculosis chemotherapy and post treatment. All patients were diagnosed as having urinary tuberculosis by positive NAAT and specific renal deformity revealed by imaging. In two of the three patients, positive culture results were obtained and one was negative in standard culture. During antituberculosis chemotherapy, a negative NAAT was obtained from 3 to 5 months after the start of treatment and no positive culture results were obtained during the same period. At the end of chemotherapy, 6 months or more after the start of medication, all patients had negative NAAT results. These results suggest that NAAT for Mycobacterium tuberculosis provides an effective and rapid detection method for urinary tuberculosis both pre- and post-treatment. PMID- 16369739 TI - Lessons learned from studies of the development of early melanoma. PMID- 16369740 TI - Early gastric cancer: concepts, diagnosis, and management. PMID- 16369741 TI - Early liver cancer: concepts, diagnosis, and management. AB - Hepatocellular carcinoma is a major health problem worldwide. The prognosis of patients depends on the stage when the disease is diagnosed. For the early detection of these tumors, both the selection of patients at high risk and effective surveillance programs are important. When the tumor is diagnosed early, patients can choose potentially curative treatment, including surgical resection, transplantation, and ablation therapy. Here, we give an overview of the present management of hepatocellular carcinoma. Further studies in diagnosis, treatment, and prevention are required to achieve the best clinical management of this disease. PMID- 16369742 TI - Early cancers of the skin: clinical, histopathological, and molecular characteristics. AB - Because skin lesions are visible and easily accessible, skin cancers provide us with an excellent in vivo model to study the development of cancers. Cutaneous malignant melanoma and squamous cell carcinoma (SCC) both arise from the epidermis and have an initial progression stage in which proliferation of the neoplastic cells is confined to the epidermis. This stage is called melanoma in situ or SCC in situ. Molecular analyses of melanoma in situ and of solar keratosis, a prototype of early SCC in situ, show that loss of p16(INK4a)/p14(ARF) and dysfunction of p53 play a critical role, respectively. Furthermore, there seems to be potential precursor cells to these in situ lesions, which are not discernible with conventional hematoxylin and eosin stained sections. The precursor cells have minimal but critical genetic alterations, such as cyclin D1 amplification and p53 mutation, and can be identified using fluorescent in situ hybridization and immunostaining with p53 antibodies, respectively. These precursor cells may be defective in repair response to DNA damage, and would have proliferative or survival advantages over their normal neighboring counterparts in the presence of growth factor stimulation or genotoxic events, such as ultraviolet irradiation. Such precursor clones may be induced at a rather young age, and their number and size increase with accumulating carcinogenic stimuli. If these lesions acquire additional mutations, they could progress to clinically visible lesions of in situ carcinoma. Precise molecular analyses of early stages of skin cancers may have a strong impact on our understanding of in vivo development of cancers in other human organs. PMID- 16369743 TI - Comparison of three radiotherapy treatment planning protocols of definitive external-beam radiation for localized prostate cancer. AB - BACKGROUND: Three radiotherapy treatment planning (RTTP) protocols for definitive external-beam radiation for localized prostate cancer, designed and clinically applied at Kyoto University, were compared. METHODS: Treatment plans were created according to three different RTTP protocols (old three-dimensional conformal radiotherapy [3D-CRT], new 3D-CRT, and intensity-modulated radiotherapy [IMRT]) on computed tomography (CT) data sets of five patients with localized prostate cancer. The dynamic-arc conformal technique was used in the 3D-CRT protocols. Differences in dose distribution were evaluated and compared based on dose-volume histogram (DVH) analyses. RESULTS: The coverage of the clinical target volume (= prostate alone) was comparable among the three RTTP protocols. However, the average values for the percent volume that received at least 95% of the prescription dose (V95), the percent of the prescription dose covering 95% of the volume (D95), and the conformity index of the planning target volume (PTV) were 99%, 97%, and 0.88 for the IMRT; 93.9%, 94.5%, and 0.76 for the new 3D-CRT; and 59.6%, 82.9%, and 0.6 for the old 3D-CRT protocol, respectively. Inhomogeneity of doses to the PTV was larger with the IMRT protocol than with the new 3D-CRT protocol. Doses to both the rectal wall and bladder wall were almost comparable with the new 3D-CRT and IMRT protocols, but were lower with the old 3D-CRT protocol, due to the lowest prescription dose and incomplete dose coverage of the PTV. CONCLUSION: The old 3D-CRT protocol could not achieve the goals for the PTV set in the IMRT protocol. The new 3D-CRT and IMRT protocols were generally comparable in terms of both the PTV coverage and normal tissue-sparing, although the IMRT protocol achieved the most conformal dose distribution to the PTV, in return for a larger, but acceptable, dose inhomogeneity. PMID- 16369744 TI - Pilot study of angiotensin II receptor blocker in advanced hormone-refractory prostate cancer. AB - BACKGROUND: We previously demonstrated that an angiotensin II receptor blocker (ARB) had the potential to inhibit cell proliferation of prostate cancer. In this study, we examined whether an ARB could elicit an antiproliferative effect on hormone-refractory prostate cancer, clinically. METHODS: Twenty-three patients with advanced hormone-refractory prostate cancer who had already received secondary hormonal therapy using dexamethasone, and who were no longer receiving conventional therapy, were enrolled. All of the patients received candesartan 8 mg once daily per os and, simultaneously, androgen ablation. Change in prostate specific antigen (PSA) was determined as the primary endpoint. The secondary end point was change in performance status (PS). To investigate angiotensin II type 1 (AT1) receptor expression in prostate cancer tissue, real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was performed, using specimens, from untreated patients with prostate cancer. RESULTS: Eight patients (34.8%) showed responsive PSA changes; six showed a decrease immediately after starting administration and two showed a stable level of PSA. Six men with a PSA decline of more than 50% showed an improvement in PS. The mean time to PSA progression (TTPP) in responders was 8.3 months (range, 1-24 months). Half of the patients showed stable or improved PS during treatment. With regard to toxic effects, only one patient showed hypotension during treatment. The RT-PCR showed that AT1 receptor expression in well-differentiated adenocarcinoma was higher than that in poorly differentiated adenocarcinoma. CONCLUSION: These data showed that an ARB had potential biological effects on prostate cancer, suggesting the usefulness of the cytostatic activity of such agents on recurrent prostate cancer. PMID- 16369745 TI - Anxiety, depression and quality of life in colorectal cancer patients. AB - BACKGROUND: Few studies have examined psychological distress and its relationship with quality of life (QL) dimensions in colorectal cancer patients. METHODS: One hundred and twenty-eight outpatients were given psychological tests for anxiety and depression (Hospital Anxiety and Depression Scale; HADS) and QL The European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire C30 (EORTC QLQ-C30) on the same occasion. The association between the patients' emotional function (EF) scoring on EORTC QLQ-C30 and their HADS scores was analyzed by multiple linear regression. RESULTS: Statistically significant negative relationships were found between EF and HADS-A (anxiety), HADS-D (depression), and HADS-T (total score), respectively, with the highest correlation coefficient being for HADS-A. However, HADS-D was significantly more highly correlated than HADS-A to other QL dimensions, and depression was more highly correlated than anxiety with reduced QL. CONCLUSION: The EF dimension of the EORTC QLQ-C30 predominantly assesses anxiety. Depression has a stronger impact on the global QL of patients than anxiety; therefore, the use of an additional instrument is recommended for the assessment of depression in outpatients with colorectal cancer. PMID- 16369746 TI - Phase I study of cisplatin and irinotecan combined with concurrent hyperfractionated accelerated thoracic radiotherapy for locally advanced non small cell lung carcinoma. AB - BACKGROUND: Irinotecan, when combined with cisplatin, is an effective treatment for advanced non-small cell lung cancer (NSCLC). This constitutes a rationale for conducting a phase I study of chemoradiotherapy including this combination for locally advanced NSCLC. PATIENTS AND METHODS: Patients with locally advanced NSCLC and a performance status of 0 or 1 were eligible. The protocol consisted of escalating doses of irinotecan on days 1 and 15, and daily low-dose cisplatin (6 mg/m(2) daily for a total dose of 120 mg/m(2)) combined with concurrent hyperfractionated accelerated thoracic irradiation (1.5 Gy twice daily for a total dose of 60 Gy). RESULTS: The maximum tolerable dose was 50 mg/m(2) of irinotecan, and the dose-limiting toxicity was esophagitis. Tumor response was observed in 50% of cases, and the median survival time of the 12 patients enrolled was 10.1 months, including two patients with 5-year disease-free survival. A pharmacokinetics study demonstrated an accumulation of total platinum, but not of free platinum, during the 26-day treatment period. CONCLUSION: The recommended dose for phase II studies was determined. PMID- 16369747 TI - Non-small cell lung cancer: radiation therapy for locoregional recurrence after complete resection. AB - BACKGROUND: We investigated patterns of failure after radical radiation therapy in relation to the radiation field in patients with postsurgical locoregional recurrence of non-small cell lung cancer. METHODS: Between 1992 and 2002, 31 patients with locoregional recurrence were treated with radiation therapy. At the time of radiation therapy, the sites of recurrence were the bronchial stump, the regional lymph nodes, the chest wall, and both the regional lymph nodes and the chest wall in 7, 20, 3, and 1 patient, respectively. The prescribed dose was 60 Gy in 30 fractions over 6 weeks in all patients. RESULTS: The response rate was 87%. The overall 1-year, 2-year, and 4-year Kaplan-Meier survival rates were 61%, 30%, and 15%, respectively, and the median survival time was 14 months. Locoregional relapse with or without distant metastasis occurred in 15 patients (in-field, 7; marginal, 7; out-field, 1), and distant metastasis alone occurred in 7 patients. The sites of marginal relapse were the upper margin in two patients, the ipsilateral margin in one patient, the contralateral margin in one patient, and the lower margin in three patients, respectively (in one patient, the data for marginal relapse overlapped). In all patients with relapse on the lower margin, the mediastinal lymph nodes were dissected at the initial surgery. CONCLUSION: Postoperative recurrent non-small cell lung cancer showed distinctive features: the response rate was high, and the incidence of marginal relapse was also high, as in small cell lung cancer. The incidence of lower marginal relapse was high, in contrast to that in surgery-naive patients. PMID- 16369748 TI - Small cell carcinoma of the esophagus responding to fourth-line chemotherapy with weekly paclitaxel. AB - A patient was diagnosed with a small cell carcinoma of the esophagus (T4N1M1b by the International Union Against Cancer [UICC] classification) in October 2002, and initially received two courses of concurrent chemotherapy with 5-fluorouracil (5-FU; 400 mg/m(2) by continuous infusion; days 1-5 and 8-12) and cisplatin (40 mg/m(2) by drip infusion; days 1 and 8) and radiation therapy (2 Gy/day, days 1 5, 8-12, and 15-19; total, 30 Gy per course) with the second course given after a 2-week interval. Two courses of chemotherapy with 5-FU (800 mg/m(2); days 1-5) and cisplatin (80 mg/m(2); day 1) given after this was completed. Although a complete response had been confirmed, recurrence with multiple liver and lymph node metastases was detected 3 months after the cessation of the second course of chemotherapy. Although the patient received second-line chemotherapy with irinotecan (150 mg/m(2); every 2 weeks) from June 2003, the disease progressed. Brain metastases developed during third-line chemotherapy with gemcitabine (1000 mg/m(2) weekly by drip infusion). The symptoms were attenuated after whole-brain radiation (30 Gy), and fourth-line chemotherapy using paclitaxel (80 mg/m(2); weekly) was initiated from November 2003. A computed tomography scan 1 month after the first course of paclitaxel showed remarkable regression of the liver metastases. The treatment strategy used for treating small cell carcinomas of the lung may be applicable for these carcinomas of the esophagus. PMID- 16369749 TI - Partial resection of the second portion of the duodenum for gastrointestinal stromal tumor after effective transarterial embolization. AB - We report herein the case of 64-year-old man with gastrointestinal stromal tumor (GIST), who was treated by partial resection of the duodenum after preoperative transarterial embolization. He presented to our hospital with a history of tarry stools, dizziness, and severe anemia (hemoglobin, 7.5 g/dl). Gastroduodenal endoscopy revealed the presence of a submucosal tumor in the second portion of the duodenum. The presence of the tumor was subsequently confirmed by double contrast gastrointestinal radiography and abdominal computed tomography. Super selective angiography showed tumor staining fed from the anterior and posterior superior pancreaticoduodenal arteries, and the inferior pancreaticoduodenal artery. Two weeks after transarterial embolization through these vessels, the tumor size was found to have shrunk to 40% of its original size. Partial resection of the duodenum was performed and absence of tumor cells at the surgical margin was confirmed by intraoperative frozen-section examination. Histopathological examination revealed that the duodenal submucosal tumor consisted of spindle cells, and immunohistochemical analysis revealed positive tumor staining for c-kit protein, CD34 and alpha-smooth muscle actin (SMA), and negative staining for desmin and S-100; the positivity rate for MIB-1 staining was 2.2%. Based on these findings, the tumor was diagnosed as a GIST of low-grade malignancy, classified as the muscular type. It is considered that preoperative treatment of duodenal GISTs, such as transarterial embolization, may be useful for reducing the extent of resection, from pancreaticoduodenenctomy to a partial resection. PMID- 16369750 TI - A new therapeutic approach in patients with advanced sarcoma. AB - Sarcomas represent a rare and heterogeneous disease and the prognosis of patients remains poor, with a disease-free survival at 5 years of less than 10%. Only a few chemotherapeutic agents, such as doxorubicin and ifosfamide, have been identified to be active with response rates above 20%. The concept of angiostatic therapy in combination with proapoptotic biomodulators and chemotherapeutics has not been evaluated in these patients. Therefore, the efficacy of low-dose trofosfamide in combination with the peroxisome proliferator-activated receptor gamma-agonist, pioglitazone, and the selective cyclooxygenase-2 inhibitor, rofecoxib, was evaluated in a pilot study. Six patients with advanced sarcoma received a combination of oral pioglitazone plus rofecoxib and, after 14 days, oral trofosfamide. The therapy was administered continuously daily. Four patients received the triple combination as maintenance therapy; three of them achieved stabilization of disease. Two patients received the combination as relapse therapy; however, it failed to stop disease progression. Side effects were generally mild and hospitalization was not necessary. This new triple combination of low-dose trofosfamide, pioglitazone, and rofecoxib may represent a feasible new alternative in the palliative treatment of sarcoma patients. PMID- 16369751 TI - 5-Fluorouracil cardiotoxicity induced by alpha-fluoro-beta-alanine. AB - Cardiotoxicity is a rare complication occurring during 5-fluorouracil (5-FU) treatment for malignancies. We herein report the case of a 70-year-old man with 5 FU-induced cardiotoxicity, in whom a high serum level of alpha-fluoro-beta alanine (FBAL) was observed. The patient, who had unresectable colon cancer metastases to the liver and lung, was referred to us for chemotherapy from an affiliated hospital; he had no cardiac history. After admission, the patient received a continuous intravenous infusion of 5-FU (1000 mg/day), during which precordial pain with right bundle branch block occurred concomitantly with a high serum FBAL concentration of 1955 ng/ml. Both the precordial pain and the electrocardiographic changes disappeared spontaneously after the discontinuation of 5-FU. As the precordial pain in this patient was considered to have been due to 5-FU-induced cardiotoxicity, the administration of 5-FU was abandoned. Instead, oral administration of S-1 (a derivative of 5-FU), at 200 mg/day twice a week, was instituted, because S-1 has a strong inhibitory effect on dihydropyrimidine dehydrogenase, which catalyzes the degradative of 5-FU into FBAL. The serum FBAL concentration subsequently decreased to 352 ng/ml, the same as the value measured on the first day of S-1 administration. Thereafter, no cardiac symptoms were observed. The patient achieved a partial response 6 months after the initiation of the S-1 treatment. The experience of this case, together with a review of the literature, suggests that FBAL is related to 5-FU-induced cardiotoxicity. S-1 may be administered safely to patients with 5-FU-induced cardiotoxicity. PMID- 16369752 TI - Successful diagnosis of thromboembolism before surgery in a woman with clear cell adenocarcinoma of the endometrium. AB - Patients with ovarian cancer with clear cell histology often have venous thromboembolism (VTE) even before surgery. In view of the possible association between clear cell histology and VTE in endometrial cancer, we measured the plasma levels of thrombin-antithrombin III complex (TAT) and D-dimer (DD) in the preoperative examinations of a patient with clear cell adenocarcinoma of the endometrium. Plasma TAT and DD were both highly elevated, though the patient had no symptoms of VTE or risk factors such as obesity or diabetes mellitus. Ultrasound Doppler examination and lung perfusion scintigraphy just before surgery revealed a thrombosis in the left popliteal vein and a pulmonary embolism. After implanting an inferior vena cava filter to prevent a fatal embolism of the lung, we performed abdominal total hysterectomy, bilateral salpingo-oophorectomy, and sampling of the pelvic lymph nodes. The VTE gradually disappeared and the plasma levels of TAT and DD returned to normal after surgery. Possibly, the VTE in this patient may have been associated with the clear cell histology. PMID- 16369753 TI - Paraganglioma of the cerebellum: case report and review of the literature. AB - Intracerebral paragangliomas are tumors with a low incidence. Only 13 cases have been reported in the literature to date. This type of tumor shows characteristic histopathological features but unclear histogenesis. Owing to its low incidence, little knowledge exists concerning the best therapeutic strategy. We report on a 55-year-old male patient with the diagnosis of a paraganglioma of the cerebellum. We compare our patient's case to similar cases reported in the literature, describe the typical course of the disease, and discuss therapeutic options. No relapse of disease was observed in our patient within 2 years of diagnosis. Surgery and adjuvant conventionally fractionated external beam radiotherapy seem to be well tolerated and feasible. A review of patients reported in the literature and a comparison to our patient reveals a good prognosis for intracerebral paragangliomas. The optimal therapeutic strategy with regard to the benefit of adjuvant radiotherapy after total or subtotal resection has yet to be defined. PMID- 16369754 TI - Nonsteroidal anti-inflammatory drugs (NSAIDs) in the perioperative phase in traumatology and orthopedics effects on bone healing. AB - OBJECTIVE: To achieve analgesic, anti-inflammatory and antipyretic effects in traumatology and orthopedic surgery without side effects or with the least possible side effects, with special emphasis on bone healing. INDICATIONS: Acute and chronic inflammatory conditions, e. g., rheumatoid arthritis, ankylosing spondylitis. Degenerative joint disease. Posttraumatic and postoperative pain, edema, or fever. Prevention of heterotopic bone formation. CONTRAINDICATIONS: Hypersensitivity. Gastrointestinal ulceration or bleeding. Severe hepatic or renal impairment. RESULTS: Nonsteroidal anti-inflammatory drugs (NSAIDs) are invaluable in treating a variety of musculoskeletal conditions. As well as their excellent analgesic potency their anti-inflammatory effects are beneficial in treating posttraumatic and postoperative edema. In addition, NSAIDs inhibit heterotopic bone formation after hip arthroplasty. Animal studies, however, have demonstrated that they cause delayed fracture healing. Although clinical studies have not yet supplied unequivocal evidence of this effect in human subjects, the authors recommend that in the presence of other risk factors which may adversely affect fracture healing, such as smoking, diabetes mellitus or peripheral arterial occlusive disease, the indication of NSAID use for analgesia should be strictly limited. Therapeutic alternatives such as centrally acting agents (e. g., weak opioids) should be considered in these patients. PMID- 16369755 TI - Posterior instrumentation of the cervical spine using the neon occipito-cervical system. Part 2: cervical and cervicothoracic instrumentation. AB - OBJECTIVE: Posterior cervical and cervicothoracic stabilization, if necessary, in combination with posterior spondylodesis, after closed or open reduction as well as with or without neural decompression as clinically indicated. INDICATIONS: Instabilities and deformities of rheumatoid, traumatic, neoplastic, infectious, iatrogenic or congenital origin. Multilevel cervical spinal stenosis with degenerative instability or kyphosis of the affected spinal segment. Corrective spondylodesis for posttraumatic or postinfectious kyphosis. CONTRAINDICATIONS: Infection in the operative field. Inability to undergo anesthesia. SURGICAL TECHNIQUE: Prone position, rigid head fixation, e. g., with Mayfield tongs. If appropriate, closed reduction under lateral image intensification. Midline posterior surgical approach at the level of the segment to be instrumented. If necessary, open reduction. Insertion of the cervical/upper thoracic screws. If necessary, posterior decompression. Instrumentation with prebent longitudinal rods. If a fusion is to be obtained, decortication of the posterior bone elements with a burr and onlay of cancellous bone. RESULTS: 44 patients, 13 women and 31 men, with an average age of 57 years were operated on between August 2000 and August 2003. All patients underwent a follow-up examination 4-39 months (average 15.7 months) after the initial surgery. Fusion was achieved in all patients who had undergone a posterior spondylodesis: no implant failure, no implant removal or reoperation. The preoperatively planned instrumentation could be implemented intraoperatively in all patients. PMID- 16369756 TI - Arthroscopic tenodesis for lesions of the long head of the biceps. AB - OBJECTIVE: Resection of the intraarticular part of the long head of the biceps and tenodesis to decrease shoulder pain due to a pathologically altered biceps tendon and to improve shoulder function. INDICATIONS: Tenosynovitis. State of prerupture. Instability (subluxation or dislocation of tendon from intertubercular groove). Intraarticular entrapment secondary to hypertrophy of the long head of the biceps in the presence of an intact cuff. To be performed during arthroscopic cuff repair or during debridement of an irreparable cuff tear. CONTRAINDICATIONS: Very thin, frayed, almost ruptured biceps tendon. Complete rupture of the long head of the biceps. SURGICAL TECHNIQUE: Standard arthroscopy with 30 degrees scope inserted through the posterior portal. Detachment of the long head from the glenoid origin. Longitudinal opening of the bicipital groove. Exteriorization and doubling of the tendon. Drilling of a socket starting in the groove but perforating the posterior cortex only with a guide wire. Passing of the tendon in an anteroposterior direction and securing the anchorage with a bioresorbable PLA interference screw. RESULTS: Between 1997 and 1999, an arthroscopic tenodesis was performed in 43 patients. Minimum follow up 2 years. The absolute Constant Score improved from 43 points preoperatively to 79 points at the time of follow-up. No loss of elbow extension or flexion; power of biceps after tenodesis 90% of opposite side. Early on, two failures of tenodesis occurred. PMID- 16369757 TI - Dorsal double-plate fixation of the distal radius. AB - OBJECTIVE: Restoration of the intra- and extraarticular anatomy of the distal radius. Stable internal fixation of fragments, with the possibility of early functional rehabilitation. INDICATIONS: Distal intraarticular radius fractures with impacted articular fragments and displaced dorsoulnar fragment. Distal intraarticular radius fractures with bony or ligamentous injury of the proximal carpal row. CONTRAINDICATIONS: General medical contraindications for surgical intervention. Distal radius fractures with palmar tilt of the distal fragment. SURGICAL TECHNIQUE: Dorsal longitudinal incision. Approach to the intermediate column via the third extensor compartment by detaching the extensor pollicis longus (EPL) tendon. Arthrotomy and revision of the proximal carpal row. Reconstruction of the radiocarpal articular surface and support with a plate applied to the dorsoulnar aspect. Approach to the radial column by subcutaneous preparation between skin flaps (cave: superficial branch of the radial nerve) and retinaculum, incision of the first extensor compartment and support of the radial column with a preshaped plate, which is pushed through under the tendons of the first compartment. Cancellous bone grafting is usually not necessary. Subcutaneous displacement of the EPL tendon with the aid of a small retinacular flap. MANAGEMENT RESULTS: Application of a removable velcro cuff. Immediate functional postoperative physiotherapy, without the cuff. No straining or forcing until first radiographic examination at 6 weeks after the operation. RESULTS: 25 consecutive patients were monitored following a double-plate fixation, with a minimum follow-up of 12 months. In all cases the reduction, in accordance with the Stewart Score, was very good, a loss of reduction was not observed. The range of motion was between 100 degrees and 160 degrees for flexion/extension and between 160 degrees und 180 degrees for pronation/supination. The average DASH Score was 7.2 points, the PRWE Score 8.0 points. No relevant loss of strength (JAMAR dynamometer) was found in any of the patients in comparison with the healthy side. Complications noted were a muscle adhesion in the region of the first extensor compartment as well as a mild reflex sympathetic dystrophy, which healed without consequences. Implants were removed from six of the patients. PMID- 16369758 TI - Endoscopic transforaminal nucleotomy with foraminoplasty for lumbar disk herniation. AB - OBJECTIVE: Removal of a sequestered lumbar disk; in instances of foraminal stenosis a decompression (foraminoplasty) can also be performed. The objective is to restore the best possible pain-free function of the spinal column. INDICATIONS: Prolapsed lumbar vertebral disk, sequestered caudally or cranially, that is no longer responding to nonoperative treatment. Cauda equina syndrome. CONTRAINDICATIONS: Sequestered herniated lumbar disk posterior to the dura mater. SURGICAL TECHNIQUE: Lateral decubitus on a radiolucent table allowing the use of an image intensifier. The usual access to a sequester at segments L5/S1 and L4/5 is approximately 12-14 cm lateral to the midline, at segments L3/4 and L2/3 approximately 10 cm lateral to the midline. In the case of sequestration in a cranial direction a somewhat smaller angle of access should be chosen, for caudal displacement the angle should be greater. Reaming of a lateral transforaminal approach to the spinal canal. The sequester is removed endoscopically through a working cannula. POSTOPERATIVE MANAGEMENT: The patient remains in the recovery room for approximately 2 h, is fitted with a brace and transferred to the ward. No medicinal thrombosis prophylaxis. Follow-up examination the next morning. Physiotherapy after 1 week. The brace is worn for approximately 2-6 weeks. MRI checkup and after 3 months evaluation of the development of strength and the state of the back musculature. RESULTS: Between January 1999 and January 2002, 611 patients underwent surgery, 32% women and 68% men, age between 18 and 65 years (average age 43.8 years). Clinical follow-up examinations were carried out after 3 months, follow-ups with questionnaire after 1 and 2 years. The evaluation criteria were based on an own questionnaire, and also on MacNab's score and the visual analog scale (VAS). After 2 years 558 patients (91.2%) responded to the questionnaires. Excellent or good results were achieved in 95.3% of the patients. 74.7% were very satisfied, 20.6% satisfied. The result was judged unsatisfactory by 4.7% of patients (less satisfied 3.9%, unsatisfied 0.8%). The numbness of the leg, present in 448 patients preoperatively, was either no longer present (63.9%) or had improved (30.3%). There were no serious complications, in particular no infections. The recurrence rate was 3.6%. PMID- 16369759 TI - Oblique upper tibial opening wedge osteotomy for genu varum. AB - OBJECTIVE: Restoration of mechanical axis of the leg. INDICATIONS: Osteoarthritis of medial knee compartment. Overload of medial compartment. Genu varum. CONTRAINDICATIONS: Smoker. Vascular impairment. Poor soft-tissue envelope. SURGICAL TECHNIQUE: Straight 12-cm midline incision starting distal to the tibial tubercle medially and continuing parallel to the tibial crest. Oblique osteotomy at 60 degrees distal-medial to proximal-lateral preserving the lateral cortex. Osteotomy wedged open and alignment checked with diathermy cord. Two tricortical bone blocks harvested from ipsilateral iliac crest or bone substitute wedges placed in osteotomy. Stabilization with contoured T-plate. Closure over drain. RESULTS: 44 patients (32 men, twelve women, average age 48 years) with 45 osteotomies. Follow-up 30 months (7-74 months). All osteotomies consolidated. 90% patients had excellent results according to the HSS (Hospital for Special Surgery) Knee Score. In 77% the femorotibial angle was corrected to 5-12 degrees . Patients regained their preoperative range of motion. There were no major complications. PMID- 16369760 TI - Quantitative trait loci for yield and related traits in the wheat population Ning7840 x Clark. AB - Grain yield and associated agronomic traits are important factors in wheat (Triticum aestivum L.) improvement. Knowledge regarding the number, genomic location, and effect of quantitative trait loci (QTL) would facilitate marker assisted selection and the development of cultivars with desirable characteristics. Our objectives were to identify QTLs directly and indirectly affecting grain yield expression. A population of 132 F12 recombinant inbred lines (RILs) was derived by single-seed descent from a cross between the Chinese facultative wheat Ning7840 and the US soft red winter wheat Clark. Phenotypic data were collected for 15 yield and other agronomic traits in the RILs and parental lines from three locations in Oklahoma from 2001 to 2003. Twenty-nine linkage groups, consisting of 363 AFLP and 47 SSR markers, were identified. Using composite interval mapping (CIM) analysis, 10, 16, 30, and 14 QTLs were detected for yield, yield components, plant adaptation (shattering and lodging resistance, heading date, and plant height), and spike morphology traits, respectively. The QTL effects ranged from 7 to 23%. Marker alleles from Clark were associated with a positive effect for the majority of QTLs for yield and yield components, but gene dispersion was the rule rather than the exception for this RIL population. Often, QTLs were detected in proximal positions for different traits. Consistent, co-localized QTLs were identified in linkage groups 1AL, 1B, 4B, 5A, 6A, and 7A, and less consistent but unique QTLs were identified on 2BL, 2BS, 2DL, and 6B. Results of this study provide a benchmark for future efforts on QTL identification for yield traits. PMID- 16369761 TI - [Secondary malignancies after successful primary treatment of malignant Hodgkin's lymphoma]. AB - Malignant Hodgkin's lymphoma (HL) has become a curable disease through the increasing intensity of the treatment strategies applied. These regimens are aggressive, including radiotherapy and chemotherapy leading to the possibility of secondary malignancies. The German Hodgkin Lymphoma Study Group considered three cohorts including 5,411 patients with all stages of HL. In 127 patients a secondary solid tumor was diagnosed (cumulative risk 2%, median follow-up 72 months), with bronchial carcinomas (23.6%) and colorectal adenocarcinomas (20.5%) being the most frequent neoplasms. Secondary acute myeloid leukemia was found in 36 patients, another ten developed myeloid dysplasia (cumulative risk 1%, median follow-up 55 months). A total of 52 patients revealed a non-Hodgkin's lymphoma (NHL; cumulative risk 0.9%, median follow-up 46 months). The overall incidence of secondary malignancies was 3.9% in patients who had been treated successfully for their HL with radio- and/or chemotherapy.A secondary NHL can be particularly difficult to be distinguished from the preceding HL. Therefore, in case of a suspected relapse, a complete histopathological work-up must be performed. PMID- 16369762 TI - [NSAID-colonopathy]. AB - The pain-relief properties of NSAID/ASA preparations have been known for a long time. In particular, the gastrointestinal tract shows side effects such as: erosion, ulceration, and even perforation. In the upper gastrointestinal tract, our group has shown that a correct histological diagnosis of NSAID/ASA induced lesions can be made in a high percentage of cases on the basis of recognition of ischemic necrosis. NSAID/ASA induced lesions are less commonly found in the lower gastrointestinal tract. We could also demonstrate a correct histological diagnosis of NSAID-colonopathy on the basis of finding ischemic necrosis. Besides the known complications, another typical complication is diaphragm-like stenosis, which must not be mistaken for tumor stenosis. The differential diagnosis of NSAID-colonopathy includes ischemic colitis, which cannot be distinguished histologically if the exact endoscopic description is not available. Sometimes NSAID/ASA induced lesions are misdiagnosed as Crohn's disease due to the focal character of the lesions. Since all of our analyses are retrospective, the criteria developed by our group should be checked prospectively. PMID- 16369763 TI - Extreme skewing of X chromosome inactivation in mothers of homosexual men. AB - Human sexual preference is a sexually dimorphic trait with a substantial genetic component. Linkage of male sexual orientation to markers on the X chromosome has been reported in some families. Here, we measured X chromosome inactivation ratios in 97 mothers of homosexual men and 103 age-matched control women without gay sons. The number of women with extreme skewing of X-inactivation was significantly higher in mothers of gay men (13/97=13%) compared to controls (4/103=4%) and increased in mothers with two or more gay sons (10/44=23%). Our findings support a role for the X chromosome in regulating sexual orientation in a subgroup of gay men. PMID- 16369764 TI - Allelic diversity in the TGFB1 regulatory region: characterization of novel functional single nucleotide polymorphisms. AB - Altered TGF-beta1 expression due to polymorphisms affects a wide variety of normal cellular and disease processes such as T cell activation and proliferation, tumor progression, and asthma. In this study, a comprehensive examination of function and diversity was undertaken for the TGFB1 promoter region and exon 1 (-2,665 to +423). The known TGF-beta1 promoter was extended to encompass 463 bases by the identification of a strong enhancer activity for a distal segment (-2,665 to -2,204). Ten novel polymorphisms and 14 novel alleles were identified. Most single nucleotide polymorphisms (SNPs) appear to be randomly associated except c.-768_-769insC and c.+74G > C and a set of five novel polymorphisms present in a single allele in persons of African descent. The TGFB1 alleles clustered into three phylogenetic groups based on the common functional SNPs c.-1347C > T (commonly known as -509C-T) and c.+29T > C (commonly known as +869T-C) suggesting three phenotypic groups. Two SNPs unique to African-Americans affect the TGFB1 regulatory region. The c.-1287G > A SNP in the promoter alters the binding affinity of two unidentified transcription factor complexes which translates into a significant difference in reporter gene expression and the c. 387C > T SNP in the 5' UTR alters the binding of Stimulating protein 1 and 3. Thus, TGFB1 possesses a highly polymorphic, extensive regulatory region that likely impacts the pathogenesis of numerous TGF-beta1 related diseases. PMID- 16369765 TI - Evidence for novel loci for late-onset Parkinson's disease in a genetic isolate from the Netherlands. AB - We studied patients with idiopathic Parkinson's disease (PD) from an isolated population in the Netherlands aiming to map gene(s) involved in PD susceptibility. A total of 109 parkinsonism patients were independently ascertained, of whom 62 presented late-onset, idiopathic PD. Genealogical research showed that 45 index cases with idiopathic PD were linked to a common ancestor, indicating familiar clustering among the patients. This strong familial clustering was highly significant (P = 0.005) when compared to random controls from the same population. We performed a genome wide scan using 382 polymorphic markers in 44 distantly related PD patients plus 112 unaffected first-degree relatives and spouses. Our genome wide association analysis (DISLAMB) revealed evidence of association at a nominal P-value < 0.01 for markers D2S2333, D4S405, D9S158, D13S153. Other regions on chromosomes 3p, 4q, 14q, 17p and 17q were found at a significance level of P < 0.05. In a follow-up study, we investigated all the positive regions using a denser marker set and a larger sample (total of 630 individuals including all late-onset PD patients). The strongest evidence for association remained for the 9q and 14q region. A significant association was found for marker D9S1838 (OR = 2.0, 95% CI 1.1-3.5, P = 0.014) and D14S65 (OR = 3.2, 95% CI 1.7-6.1, P < 0.001). Moreover, a common haplotype with excess of sharing among late-onset PD cases was observed on both regions. Our results suggest the existence of two loci influencing PD susceptibility on chromosome 9q and 14q. PMID- 16369766 TI - Regulation of growth of Lilium plantlets in liquid medium by application of paclobutrazol or ancymidol, for its amenability in a bioreactor system: growth parameters. AB - Effect of growth retardants (paclobutrazol or ancymidol) was studied in Lilium plantlets growing in liquid culture. A significant increase in leaf chlorophyll, epicuticular wax, plant dry weight and bulb starch contents were found in plantlets treated with growth retardants. A similar increase in the number of leaves, roots and bulbs was also noted. However, total leaf area and the fresh weight increased only marginally. These features resulted in robust plantlets that showed significantly improved ex vitro survival. Based on these features, a comprehensive index (CI) was calculated as a measure of quality of the plantlets, and it correlated well with their ex vitro survival. Treatment of plantlets with 3.4 microM paclobutrazol was found to be the best and its carry over effects were also minimal. PMID- 16369767 TI - The maize Knotted1 gene is an effective positive selectable marker gene for Agrobacterium-mediated tobacco transformation. AB - We have assessed the use of a homeobox gene knotted1 (kn1) from maize as a selectable marker gene for plant transformation. The kn1 gene under the control of cauliflower mosaic virus 35S promoter (35S::kn1) was introduced into Nicotiana tabacum cv. Xanthi via Agrobacterium-mediated transformation. Under nonselective conditions (without antibiotic selection) on a hormone-free medium (MS), a large number of transgenic calli and shoots were obtained from explants that were infected with Agrobacterium tumefaciens LBA4404 harboring the 35S::kn1 gene. On the other hand, no calli or shoots were produced from explants that were infected with an Agrobacterium strain harboring pBI121 (nptII selection) or from uninfected controls cultured under identical conditions. Relative to kanamycin selection conferred by nptII, the use of kn1 resulted in a 3-fold increase in transformation efficiency. The transgenic status of shoots obtained was confirmed by both histochemical detection of GUS activity and molecular analysis. The results presented here suggest that kn1 gene could be used as an effective alternative selection marker with a potential to enhance plant transformation efficiency in many plant species. With kn1 gene as a selection marker gene, no antibiotic-resistance or herbicide-resistance genes are needed so that potential risks associated with the use of these traditional selection marker genes can be eliminated. PMID- 16369768 TI - Transformation of Montmorency sour cherry (Prunus cerasus L.) and Gisela 6 (P. cerasus x P. canescens) cherry rootstock mediated by Agrobacterium tumefaciens. AB - Sour cherry (Prunus cerasus L.) scion cv. Montmorency and rootstock cv. Gisela 6 (P. cerasus x P. canescens) were transformed using Agrobacterium tumefaciens strain EHA105:pBISN1 carrying the neomycin phosphotransferase gene (nptII) and an intron interrupted ss-glucuronidase (GUS) reporter gene (gusA). Whole leaf explants were co-cultivated with A. tumefaciens, and selection and regeneration of transformed cells and shoots of both cultivars was carried out for 12 weeks on selection medium containing 50 mg l(-1) kanamycin (Km) and 250 mg l(-1) timentin. These media were [Quoirin and Lepoivre (Acta Hortic 78:437-442, 1977)] supplemented with 0.5 mg l(-1) benzylaminopurine (BA) + 0.05 mg l(-1) indole-3 butyric acid (IBA), and woody plant medium [Lloyd and McCown (Proc Int Plant Prop Soc 30:421-427, 1980)] containing 2.0 mg l(-1) BA + 1.0 mg l(-1) IBA for cv. Montmorency and cv. Gisela 6, respectively. Seven out of 226 (3.1%) explants of cv. Montmorency and five out of 152 (3.9%) explants of cv. Gisela 6 produced 30/39 GUS- and PCR-positive shoots from the cut midribs via an intermediate callus. Southern analysis of the GUS- and PCR-positive transformants confirmed stable integration of the transgenes with 1-3 copy numbers in the genomes of seven lines of cv. Montmorency and five of cv. Gisela 6. The selected transformants have a normal phenotype in vitro. PMID- 16369769 TI - Contractile responses to selective phosphodiesterase inhibitors following chronic beta-adrenoreceptor activation. AB - Contractile responses to phosphodiesterase (PDE) inhibitors are attenuated in heart failure, an effect limiting the clinical value of these agents. In this study, we sought to determine whether abnormalities in the beta-adrenoreceptor (beta-AR)-cyclic adenosine monophosphate (cAMP) signal transduction are sufficient to account for downregulation of PDE inhibitor-induced inotropic responses following chronic sympathetic activation. Sustained beta-AR activation produced by administration of isoproterenol (ISO) (50 microg kg(-1) day(-1) i.p. for 1 month) to rats resulted in cardiac hypertrophy, but did not affect baseline cardiac systolic function, as assessed in vivo by echocardiography and ex vivo under controlled loading conditions and heart rate (left ventricular systolic pressure-volume and stress-strain relations). Moreover, chronic ISO administration did not alter the baseline myocardial norepinephrine release or inotropic responses to incremental concentrations of Ca(2+) in isolated, perfused heart preparations. However, left ventricular contractile responses to ISO, the PDE III inhibitor amrinone, and the PDE IV inhibitor rolipram were attenuated following chronic beta-AR activation. Myocardial cAMP concentrations after stimulation with amrinone and rolipram were similar in ISO-treated and control rats. However, in ISO-treated rats, a marked decrease in contractile responsiveness to the cell-permeable, PDE-resistant cAMP analogue, 8 bromoadenosine cAMP, was noted. In conclusion, these data suggest that in cardiac disease, sustained beta-AR activation, without producing ventricular systolic dysfunction or enhanced myocardial norepinephrine release, is sufficient to account for the downregulation of contractile responses to PDE inhibitors. This effect appears to be largely mediated through abnormalities in signal transduction between cAMP and Ca(2+)-induced Ca(2+) release. PMID- 16369770 TI - Effects of reduced glycogen on structure and in vitro function of rat sarcoplasmic reticulum Ca2+-ATPase. AB - The aim of this study was to examine the effects of reduced glycogen concentration on sarcoplasmic reticulum (SR) Ca(2+)-ATPase activity in rat fast twitch muscles. In the first experiment, the gastrocnemius (GAS) muscle from one leg was removed, followed by starvation for 24-72 h, after which the remaining GAS was removed. Intra-animal comparisons revealed that starvation caused a 25% reduction (P<0.05) in the glycogen concentration but no change in SR Ca(2+) ATPase activity in the GAS. In the second experiment, the SR was purified from a mixture of the GAS and vastus lateralis muscles. In half of the samples obtained from each animal, glycogen was extracted from the SR by treatment with glucoamylase. Treatment resulted in a 94.1 and 70.2% decrease (P<0.01) in glycogen and glycogen phosphorylase, respectively, and a 41.5% increase (P<0.05) in a fluorescein isothiocyanate (FITC) binding to SR Ca(2+)-ATPase. On the other hand, SR Ca(2+)-ATPase activity and the affinity of the enzyme for ATP were unaltered. These results do not implicate depletion of muscle glycogen as a contributor to impaired SR Ca(2+)-ATPase activity as measured in vitro. Therefore, it is concluded that muscle glycogen does not influence exercise tolerance and work productivity in working muscles by modulating the structure of protein involved in Ca(2+) sequestering. Furthermore, it is suggested that the FITC binding assay may be inappropriate as a method for examining the mechanisms for the altered activity of SR Ca(2+)-ATPase. PMID- 16369771 TI - Inhibition of membrane depolarisation-induced transcriptional activity of cyclic AMP response element binding protein (CREB) by the dual-leucine-zipper-bearing kinase in a pancreatic islet beta cell line. AB - AIMS/HYPOTHESIS: The activation of the transcription factor cyclic AMP response element binding protein (CREB) by protein kinase A is inhibited by the human orthologue of the mitogen-activated protein kinase, dual-leucine-zipper-bearing kinase (DLK) in teratocarcinoma cells. However, pancreatic beta cells are electrically excitable and a major pathway regulating CREB in these cells is membrane depolarisation, leading to calcium influx and activation of the calcium/calmodulin-dependent protein phosphatase calcineurin. Therefore, the effect of DLK on CREB activity induced by membrane depolarisation was investigated in the beta cell line HIT. MATERIALS AND METHODS: Reporter gene assays and biochemical techniques were used. RESULTS: RT-PCR, Western blot analysis and immunohistochemistry demonstrated the expression of DLK in HIT cells and primary mouse islets. In transient transfection experiments, DLK inhibited both GAL4-CREB activity induced by membrane depolarisation, and transcription directed by the CREB binding site, the cyclic AMP response element. Furthermore, DLK inhibited the transcriptional activity conferred by the CREB coactivator, CREB binding protein, both under basal conditions and after membrane depolarisation. DLK was also effective in response to glucose, the most potent physiological stimulus and known to cause membrane depolarisation of beta cells. Inhibition of calcineurin enhanced DLK activity, whereas overexpression of calcineurin reduced the inhibition by DLK of transcription directed by cyclic AMP response element after membrane depolarisation. CONCLUSIONS/INTERPRETATION: These results demonstrate a calcineurin-sensitive inhibition by DLK of CREB activity after membrane depolarisation in pancreatic islet beta cells. This inhibition may, at least partially, be mediated at the coactivator level. The results thus suggest that DLK plays a role in the regulation of beta cell function, including insulin gene transcription and beta cell apoptosis. PMID- 16369772 TI - Expression and function of leukaemia inhibitory factor and its receptor in normal and regenerating rat pancreas. AB - AIMS/HYPOTHESIS: It was recently reported that culturing adult exocrine cells in the presence of epidermal growth factor and leukaemia inhibitory factor (LIF) resulted in their transdifferentiation into endocrine beta cells. The aim of this study was to examine the expression and function of LIF in the pancreas. MATERIALS AND METHODS: We studied the expression of LIF and its receptor components, LIF-receptor-beta and gp130, by immunohistochemistry, western blotting and RT-PCR in normal rat pancreas, pancreas with duct ligation-induced islet neogenesis, and in pancreatic cell cultures. Isolated duct fragments were cultured in the presence of LIF and a janus kinase 2 (JAK2) inhibitor. RESULTS: LIF was detected by immunohistochemistry, western blot and RT-PCR in the ducts of the normal pancreas. Both LIF-receptor-beta and gp130 were detected by RT-PCR in the pancreas. Immunostaining revealed gp130 exclusively in the ducts and centro acinar cells. After duct ligation-induced tissue injury, upregulation of LIF and its receptor occurred in rat pancreas. Metaplastic exocrine cells also started to express LIF and this was increased after alloxan treatment. Signalling via LIF receptor-beta/gp130 involves the JAK/signal transducer and activator of transcription (STAT) pathway. LIF induced increased activation of STAT3 in pancreatic cells. In isolated duct fragments, addition of LIF resulted in a significant increase in duct cell proliferation, while a specific inhibitor of the JAK/STAT signalling pathway inhibited proliferation. CONCLUSION/INTERPRETATION: Our observations show that LIF and its receptor are expressed in cells from pancreatic ducts. The cytokine plays a role in pancreatic physiology, controls duct cell proliferation and is involved in repair processes following pancreatic injury. PMID- 16369773 TI - Diabetes mellitus and impaired fasting glucose in Alaska Eskimos: the Genetics of Coronary Artery Disease in Alaska Natives (GOCADAN) study. AB - AIMS/HYPOTHESIS: We aimed to: (1) define the prevalence of type 2 diabetes and IFG in Eskimos in Norton Sound, Alaska; (2) determine correlates of prevalent diabetes in this population; and (3) compare the prevalence of diabetes in the Genetics of Coronary Artery Disease in Alaska Natives (GOCADAN) Study with other samples of Eskimos, Inuit, American Indians and US blacks, whites and Mexican Americans. METHODS: The GOCADAN Study enrolled 1,214 participants >or=18 years who were members of extended pedigrees from the Norton Sound region of Alaska. Diagnosed type 2 diabetes was based on reported use of insulin or hypoglycaemic medications and a medication inventory. Fasting glucose measurements were obtained to ascertain IFG status and undiagnosed diabetes according to American Diabetes Association (ADA) criteria. OGTTs were performed to ascertain diabetes according to the World Health Organization (WHO) definition. We used logistic regression analysis to model factors that were significantly associated with odds of prevalent ADA diabetes. RESULTS: The prevalences of ADA diabetes and IFG were 3.8% (5.0% of women; 2.2% of men) and 15.6% (13.9% of women; 17.7% of men), respectively. In the subset of 787 participants who took the OGTT, the prevalences of ADA and WHO diabetes were 5.1 and 6.9%, respectively. The adjusted odds of ADA diabetes was 2.8 times higher in participants meeting Adult Treatment Panel III criteria for abdominal obesity than in those who did not. The statistically significant sex-related difference in diabetes prevalence did not persist in multivariable analyses. CONCLUSIONS/INTERPRETATION: Alaska Eskimos have a low prevalence of type 2 diabetes. The high prevalence of IFG indicates that diabetes may become increasingly problematic in this population. Abdominal obesity in women may help explain why diabetes prevalence differs according to sex. PMID- 16369774 TI - Elevations of plasma methylarginines in obesity and ageing are related to insulin sensitivity and rates of protein turnover. AB - AIMS/HYPOTHESIS: Increased circulating methylarginines (MA) have been linked to the metabolic syndrome to explain endothelial dysfunction and cardiovascular disease risk. Proteins that contain MA are regulatory and release them during catabolism. We hypothesised that increased protein turnover in insulin-resistant states contributes to an increase in circulating MA. MATWERIALS AND METHODS: We performed hyperinsulinaemic, euglycaemic, and isoaminoacidaemic experiments on 49 lean, obese and elderly subjects, with measurements of the kinetics of glucose and protein metabolism. Plasma MA, i.e. asymmetrical dimethylarginine (ADMA), symmetrical dimethylarginine (SDMA), and N -monomethyl-L-arginine (NMMA), lipids and body composition were measured. RESULTS: Insulin resistance of glucose and protein metabolism occurred in obese and elderly subjects. ADMA concentrations were 29 to 120% higher in obese and 34% higher in elderly than in lean subjects. SDMA were 34 and 20% higher in obese than in lean and than in elderly subjects, respectively. NMMA were 32% higher in obese than in lean subjects. ADMA differed by sex, being higher in men, namely by 1.75x in obese men and by 1.27x in elderly men. Postabsorptive ADMA (r=0.71), SDMA (r=0.46), and NMMA (r=0.31) correlated (all p<0.05) with rates of protein flux. All three MA correlated negatively with clamp glucose infusion rates and uptake (p<0.001). ADMA and SDMA correlated negatively with net protein synthesis and clamp amino acid infusion rates (p<0.05). All MA also correlated with adiposity indices and fasting insulin and triglycerides (p<0.05). CONCLUSIONS/INTERPRETATION: Obesity, sex and ageing affect MA. Elevations of the three MA in obese, and of ADMA in elderly men, are related to increased protein turnover and to lesser insulin sensitivity of protein metabolism. These interrelationships might amplify insulin resistance and endothelial dysfunction. PMID- 16369775 TI - PAF antagonist BN-52021 reduces intercellular adhesion molecule-1 expression and oxidative stress in rats with reperfusion damage due to unilateral testicular torsion. AB - The aim of this study was to determine the effects of specific platelet activating factor (PAF) antagonist BN-52021 on intercellular adhesion molecule-1 (ICAM) expression and oxidative stress in rats with reperfusion damage due to unilateral testicular torsion. Thirty male Sprague-Dawley rats were separated into three groups, each containing ten rats. A sham operation was performed in group 1 (control). In group 2 [ischemia-reperfusion (I-R)/untreated], 1-h detorsion of the testis was performed after 6 h of unilateral testicular torsion. In group 3 (I-R/BN-52021), after performing the same surgical procedures as in groups II, BN-52021 was given intravenously at the starting time of reperfusion. In all experimental rats, ipsilateral orchiectomies were performed for histological examination and measuring the tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH Px). MDA values and the testicular injury score decreased and SOD, CAT and GSH-Px values increased in the I-R/BN-52021 treated group compared to in the I R/untreated group. Most of the specimens in the I-R/BN-52021 treated group showed grade-I testicular injury. However, the injuries in the I-R/untreated rats varied between grades III and IV. An ICAM-1 expression was intensive in the interstitial spaces and basement membrane of the tubuli seminiferi, of testicular tissue in the I-R/untreated group. However, an ICAM-1 expression was mild in the I-R/BN 52021 group. BN-52021 may play an important role in the immunohistochemical expression of adhesion molecule ICAM-1 and may reduce oxidative stress in rats with reperfusion damage due to unilateral testicular torsion. PMID- 16369776 TI - Sonic hedgehog expression in the development of hindgut in ETU-exposed fetal rats. AB - Sonic hedgehog (Shh) has been shown to be involved in the morphogenesis of many organ systems including the notochord, floor plate and limbs, as well as in the development of the left-right axis in vertebrates. Recent evidence suggests the Shh cascade plays a crucial role in the development of the foregut and hindgut. We have previously shown that prenatal exposure of fetal rats to ethylenethiourea (ETU) induces hindgut malformations and other abnormalities of the VACTERL association. The aim of this study was to determine the pattern of expression of Shh and its downstream genes during hindgut development in ETU-exposed embryos with anorectal malformations (ARMs). Pregnant Sprague-Dawley rats were mated together overnight and a positive vaginal plug was marked as gD0. On gD10, 1% ETU (125 mg/kg) was given to the experimental group and controls received the same volume of saline. Embryos were collected from both groups at gD12-16. The developing hindgut of each embryo was dissected under magnification and snap frozen. Highly purified RNA was isolated from each hindgut and first strand cDNA was prepared with appropriate negative controls. Reverse transcriptase (RT) polymerase chain reaction (PCR) was done to determine the transcripts of Shh in each sample and quantitative real-time PCR was carried out to show relative quantitative expression of Shh at each time point. Shh was detected in all samples confirming that Shh is active during the process of hindgut development in fetal rats. Relative quantitation demonstrated that Shh expression shows time dependent changes in the developing hindgut of ETU-exposed rat embryos, and when results were compared with control samples, there was significant decrease in expression on gD14 and 15, when the cloaca normally separates into the rectum and urethra occurs in the rat fetus. The misregulated expression of Shh in the hindgut of ETU-exposed rat embryos suggests that ETU may interfere with Shh signalling. Downregulation at the time of cloacal separation into rectum and urethra indicates that Shh plays a crucial role in the development of hindgut. PMID- 16369777 TI - Colocalization between caveolin isoforms in the intestinal smooth muscle and interstitial cells of Cajal of the Cav1(+/+) and Cav1 (-/-) mouse. AB - Confocal microscopic images were obtained from the immunohistochemical sections of jejeunum to determine the localization/colocalization between caveolin-1, caveolin-2 and caveolin-3 in intestinal smooth muscle cells (SMCs) and interstitial cells of Cajal (ICC) of Cav1(+/+) and Cav1(-/-) mouse. Intestinal regions were segmented [inner circular muscle (icm), outer circular muscle (ocm), myenteric plexus region (mp), and longitudinal muscle (lm)] by LSM 5 and analyzed by ImageJ to show Pearson's correlation (r (p)) and overlap coefficient (r) of colocalization. In the intestine of Cav1(+/+), caveolin-1 (cav1) was colocalized with caveolin-2 (cav2) and caveolin-3 (cav3). Cav2 also was well colocalized with cav3. In the intestine of Cav1(-/-), cav1 and cav2 were absent in all images, but reduced cav3 was expressed in ocm. Caveolae were present in cell types with cav1 in Cav1(+/+), and present with cav3 in ocm of Cav1(-/-). C-kit occurred in deep muscular plexus (ICC-DMP) and myenteric plexus (ICC-MP), in both Cav1(+/+) and Cav1(-/-), and colocalized with cav1 and cav2 in the intestine of Cav1(+/+). Cav3 was absent/present at low immunoreactivity in ICC-DMP and ICC-MP of the intestines of Cav1(+/+) and Cav1(-/-). To conclude, cav1 is necessary for the expression of cav2 in SMC and ICC of intestine and facilitates, but is not necessary for the expression of cav3. PMID- 16369778 TI - Molecular analysis of halophilic bacterial community for high-rate denitrification of saline industrial wastewater. AB - A denitrification system for saline wastewater utilizing halophilic denitrifying bacteria has not been developed so far. In this study, denitrification performance and microbial community under various saline conditions were investigated using denitrifying sludge acclimated under low-salinity condition for a few years as seed sludge. A continuous denitrification experiment showed that denitrification performance and microbial community at 10% salinity was higher than that at 1% salinity. The microbial community in the denitrification sludge that was acclimated under low salinity was monitored by terminal restriction fragment length polymorphism (T-RFLP) analysis during acclimation to high-salinity condition. T-RFLP profiles and clone analysis based on 16S rRNA encoding genes in the sludge of the denitrification system with 10% salinity indicated that the gamma-Proteobacteria, particularly Halomonas spp., were predominant species, suggesting that these bacterial members were possibly responsible for a high denitrification activity under high-salinity conditions. Furthermore, the investigation of denitrification performance under various saline conditions revealed that 4-10% salinity results in the highest denitrification rate, indicating that this salinity was optimal for predominant bacterial species to exhibit denitrification activity. These results indicate the possibility that an appropriate denitrification system for saline wastewater can be designed using acclimated sludge with a halophilic community. PMID- 16369779 TI - Autodisplay: efficient bacterial surface display of recombinant proteins. AB - To display a protein or peptide with a distinct function at the surface of a living bacterial cell is a challenging exercise with constantly increasing impact in many areas of biochemistry and biotechnology. Among other systems in Gram negative bacteria, the Autodisplay system provides striking advantages when used to express a recombinant protein at the surface of Escherichia coli or related bacteria. The Autodisplay system has been developed on the basis of and by exploiting the natural secretion mechanism of the AIDA-I autotransporter protein. It offers the expression of more than 10(5) recombinant molecules per single cell, permits the multimerization of subunits expressed from monomeric genes at the cell surface, and allows, after transport of an apoprotein to the cell surface, the incorporation of an inorganic prosthetic group without disturbing cell integrity or cell viability. Moreover, whole cells displaying recombinant proteins by Autodisplay can be subjected to high-throughput screening (HTS) methods such as ELISA or FACS, thus enabling the screening of surface display libraries and providing access to directed evolution of the recombinant protein displayed at the cell surface. In this review, the application of the Autodisplay system for the surface display of enzymes, enzyme inhibitors, epitopes, antigens, protein and peptide libraries is summarised and the perspectives of the system are discussed. PMID- 16369780 TI - Reversible acute methotrexate leukoencephalopathy: atypical brain MR imaging features. AB - BACKGROUND: Unusual acute symptomatic and reversible early-delayed leukoencephalopathy has been reported to be induced by methotrexate (MTX). OBJECTIVE: We aimed to identify the occurrence of such atypical MTX neurotoxicity in children and document its MR presentation. MATERIALS AND METHODS: We retrospectively reviewed the clinical findings and brain MRI obtained in 90 children treated with MTX for acute lymphoblastic leukaemia or non-B malignant non-Hodgkin lymphoma. All 90 patients had normal brain imaging before treatment. In these patients, brain imaging was performed after treatment completion and/or relapse and/or occurrence of neurological symptoms. RESULTS: Of the 90 patients, 15 (16.7%) showed signs of MTX neurotoxicity on brain MRI, 9 (10%) were asymptomatic, and 6 (6.7%) showed signs of acute leukoencephalopathy. On the routine brain MRI performed at the end of treatment, all asymptomatic patients had classical MR findings of reversible MTX neurotoxicity, such as abnormal high intensity areas localized in the deep periventricular white matter on T2-weighted images. In contrast, the six symptomatic patients had atypical brain MRI characterized by T2 high-intensity areas in the supratentorial cortex and subcortical white matter (n=6), cerebellar cortex and white matter (n=4), deep periventricular white matter (n=2) and thalamus (n=1). MR normalization occurred later than clinical recovery in these six patients. CONCLUSIONS: In addition to mostly asymptomatic classical MTX neurotoxicity, MTX may induce severe but reversible unusual leukoencephalopathy. It is important to recognize this clinicoradiological presentation in the differential diagnosis of acute neurological deterioration in children treated with MTX. PMID- 16369781 TI - Interaction of involuntary post-contraction activity with locomotor movements. AB - Involuntary post-contraction muscle activity may occur after performing a strong long-lasting (about 30 s) isometric muscle contraction (Kohnstamm phenomenon). Here we examined how this putative excitatory state may interact with a locomotor movement. The subjects stood upright and were asked to oppose a rotational force applied to the pelvis for about 30 s either in the clockwise or in the counterclockwise direction. After that, they were asked to perform various motor tasks with the eyes closed. During quiet standing, we observed an involuntary post-contraction torsion of the trunk. During walking, the post-contraction facilitatory effect of body torsion was not overridden by the voluntary activity, but instead significantly influenced the forward locomotor program such that subjects walked along a curved trajectory in the direction of the preceding torsion. In contrast, we did not observe any rotational component when subjects were asked to step in place. We conclude that the post-contraction rotational aftereffect does not transfer to just any motor task but apparently manifests itself in those movements that incorporate the activated axial muscle synergy or rotational component. We argue that central excitability changes following the voluntary effort may contribute to the phenomenon and highlight the role of tonic influences in fine-tuning of the spinal cord. PMID- 16369782 TI - Immunoreactivity for calretinin and calbindin in the vestibular nuclear complex of the monkey. AB - Immunoreactivity to calcium-binding proteins has been a useful extension to cytoarchitectonics in defining the organization of many central nervous system regions. Previously we found subdivisions of the cat medial vestibular nucleus (MVe) defined by immunoreactivity to the calcium-binding proteins, calretinin and calbindin. Here we report similar subdivisions in both the squirrel and the macaque monkey. Calretinin immunoreactivity reveals a small area of cells and processes located dorsally in the MVe. In the anterior-posterior direction these cells extend over less than half of the nucleus. This area is not distinct in Nissl-stained sections. Elsewhere in the vestibular nuclear complex (VNC) and in the nucleus prepositus hypoglossi (PrH) there are scattered labeled cells. Immunoreactivity for calbindin shows a small patch of dense fiber label at the border of MVe and PrH, and a patchy distribution in the rest of the VNC that changes at different anterior-posterior levels. There are also calbindin-labeled cells in the underlying reticular formation over a very restricted anterior posterior extent in both squirrel and macaque monkey. The dendrites of some of these cells can be followed into PrH, and data from other studies suggests that they may contribute to vestibular-oculomotor function. Scattered cells in the VNC are densely outlined by calbindin-labeled terminals, suggesting a major drive from the calbindin-labeled fiber input. These findings, along with observations from rodents and cats, suggest that there are subdivisions of the MVe defined by calcium-binding proteins that are homologous across rodents, cats, and New World and Old World monkeys. PMID- 16369783 TI - Perceived versus actual head-on-trunk orientation during arm movement control. AB - Static roll head tilt induces bias in the trajectory of upper limb voluntary movements. The aim of the experiment was to investigate whether this bias is dependant on the perception of body configuration rather than on its actual configuration. We used the 'return' phenomenon as a method to produce dissociation between perceived and actual head tilt. Static roll head tilt in supine subjects was sustained for 15 min during which subjects were periodically required to estimate verbally the tilt of their head respective to their trunk and draw, with their right index finger, straight lines aligned with their trunk. After 15 min, subjects' head were realigned with the trunk, and subjects continued to give verbal estimate of head position and perform the motor task. Results showed that the initial angular deviation of the lines in the direction opposite to head tilt gradually diminished. The adaptation was noticeable within the first 3-5 min of tilt and subsequently diminished. Verbal estimates confirmed the return phenomenon, i.e. subjects perceived their head as slowly returning towards its neutral position after a few minutes of sustained tilt. When realigned with the trunk, subjects experienced the illusion that their head was tilted in the opposite direction to the initial head tilt and a line deviation in the opposite direction to those made on initial exposure was observed (after effect). These results indicate that the angular deviation in motor production observed in condition of static head tilt were largely related to the perceived body configuration and therefore favour the hypothesis that the conscious perception of body configuration plays a key role in organising sensorimotor tasks. PMID- 16369784 TI - Long-term potentiation of evoked presynaptic response at CA3-CA1 synapses by transient oxygen-glucose deprivation in rat brain slices. AB - Physiological activity-dependent long-term changes in synaptic transmission, as long-term potentiation (LTP) are thought to be the substrate of learning and memory. However, a form of postsynaptic pathological LTP at the CA3-CA1 synapses has been demonstrated following few minutes of anoxia and aglycemia in vitro. The ischemia LTP shared many molecular mechanisms with the physiological LTP, and was believed to be involved in the delayed neuronal death following ischemia. However, the role of the presynaptic component in this regard is not known. Here we show that a short period of oxygen-glucose deprivation can induce a form of LTP (lasting for hours) of the presynaptic response at the CA3-CA1 synapses. This form of LTP is independent of postsynaptic alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors, but Ca(2+) dependent. This presynaptic LTP may represent a presynaptic hyperexcitability of the afferent fibers following ischemia, and responsible for the excitotoxicity to the CA1 neurons (ischemia-induced increases of glutamate release that kills neurons) and the postsynaptic pathological ischemic LTP. PMID- 16369785 TI - Task-switching with antisaccades versus no-go trials: a comparison of inter-trial effects. AB - Antisaccades involve the suppression of a pre-potent prosaccade and a vector inversion to generate the novel ocular motor response of looking away from the target. Antisaccades have also been found to prolong the latencies of saccades in upcoming trials, an effect that we attribute to a form of immediate plasticity in the ocular motor system. Our goal was to determine whether the inter-trial effects of antisaccades were similar to that of no-go trials, where subjects must suppress making a saccade when the target appears without substituting a novel ocular motor response. We tested 12 subjects with two different blocks of saccadic trials. In one, prosaccades randomly alternated with antisaccades. In the other, prosaccades alternated with no-go trials. We analyzed the error rates and latencies of prosaccades that followed antisaccades versus no-go trials, compared to repeated prosaccades, to determine if inter-trial effects were present for both types of responses that required prosaccade suppression. No-go responses increased the error rates of prosaccades in the following trial less than antisaccades did. However, no-go trials had the same effect on the latencies of upcoming prosaccades as antisaccades. The inhibitory effect that prolongs the latencies of prosaccades after antisaccades likely stems from the need to inhibit a prosaccade, a function that is also required in no-go trials. The greater impairment of prosaccade accuracy after an antisaccade may reflect either additional control mechanisms involved in vector inversion or a different form of inhibitory control that operates during antisaccades and not during no-go responses. PMID- 16369786 TI - Neural responses in multiple basal ganglia regions following unilateral dopamine depletion in behaving rats performing a treadmill locomotion task. AB - To investigate basal ganglia (BG) neural responses to dopamine (DA) depletion, multiple channel, single unit recording was carried out in freely moving rats performing a treadmill locomotion task. Single unit activity from 64 microelectrodes in the striatum (STR), globus pallidus (GP), subthalamic nucleus (STN) and substantia nigra pars reticulata (SNr) was recorded simultaneously before and after a unilateral DA lesion induced by microinjection of 6 hydroxydopamine (6-OHDA) into the medial forebrain bundle. The DA lesion resulted in an impairment of treadmill walking manifested by a significant decrease in swing time of both forelimbs. The stance time, however, increased significantly only in ipsilateral (good) forelimbs, reflecting compensatory changes in the good limb for motor deficits. Neural activity in the STR and GP ipsilateral to the lesion decreased during the 7-day period following the DA lesion. Conversely, an increase in spike discharges appeared in the ipsilateral SNr and STN several days after the DA lesion. Changes in the type of neural response associated with treadmill locomotion were also found in some neurons after DA depletion. Such changes were most prominent in the STR. Limb movement-related neural activity increased significantly mainly in the SNr. Additionally, neural responses to the tone cue associated with the onset of the treadmill diminished greatly in the lesioned side of the BG. Increased activity in SNr neurons is consistent with the concept that inhibition of thalamus contributes to hypokinesis in the absence of DA. Substantial decrease in striatal activity supports a concept that DA loss leads to a global suppression of recurrent cortical striatal thalamic activity that degrades normal information flow in Parkinson's diseases. PMID- 16369787 TI - The brain's fingers and hands. AB - The brain keeps track of the changing positions of body parts in space using a spatial body schema. When subjects localise a tactile stimulus on the skin, they might either use a somatotopic body map, or use a body schema to identify the location of the stimulation in external space. Healthy subjects were touched on the fingertips, with the hands in one of two postures: either the right hand was vertically above the left, or the fingers of both hands were interwoven. Subjects made speeded verbal responses to identify either the finger or the hand that was touched. Interweaving the fingers significantly impaired hand identification across several experiments, but had no effect on finger identification. Our results suggest that identification of fingers occurs in a somatotopic representation or finger schema. Identification of hands uses a general body schema, and is influenced by external spatial location. This dissociation implies that touches on the finger can only be identified with a particular hand after a process of assigning fingers to hands. This assignment is based on external spatial location. Our results suggest a role of the body schema in the identification of structural body parts from touch. PMID- 16369788 TI - A dynamic fMRI study of illusory double-flash effect on human visual cortex. AB - Functional MRI (fMRI) combined with the paired-stimuli paradigms (referred as dynamic fMRI) was used to study the "illusory double-flash" effect on brain activity in the human visual cortex. Three experiments were designed. The first two experiments aimed to examine the cross-modal neural interaction between the visual and auditory sensory systems caused by the illusory double-flash effect using combined auditory (beep sound) and visual (light flash) stimuli. The fMRI signal in the visual cortex was significantly increased in response to the illusory double flashes compared to the physical single flash when the inter stimuli delay between the auditory and visual stimuli was 25 ms. This increase disappeared when the delay was prolonged to approximately 300 ms. These results reveal that the illusory double-flash effect can significantly affect the brain activity in the visual cortex, and the degree of this effect is dynamically sensitive to the inter-stimuli delay. The third experiment was to address the spatial differentiation of brain activation in the visual cortex in response to the illusory double-flash stimulation. It was found that the illusory double flash effect in the human visual cortex is much stronger in the periphery than the fovea. This finding suggests that the periphery may be involved in high-level brain processing beyond the retinotopic visual perception. The behavioral measures conducted in this study indicate an excellent correlation between the fMRI results and behavioral performance. Finally, this work demonstrates a unique merit of fMRI for providing both temporal and spatial information regarding cross modal neural interaction between different sensory systems. PMID- 16369789 TI - Determinants guiding alternate foot placement selection and the behavioral responses are similar when avoiding a real or a virtual obstacle. AB - In this study we validate the use of a virtual planar obstacle paradigm to study the avoidance of a real obstacle, such as a hole, during locomotion. Also we further validate the economy determinant implicated with the minimization of foot displacement from its normal landing position during alternate foot placement. Participants were asked to perform two blocks of trials: real (a real hole was embedded in the pathway and participants were requested to avoid stepping into it) and virtual (a virtual planar obstacle was displayed on the screen of a liquid crystal display monitor). Trunk and feet kinematics were monitored, as well as electromyography (EMG) activity of 14 muscles of both lower limbs and trunk. The results of this study showed that the dominant choice for each obstacle investigated was not different between the real and virtual conditions. In addition, dynamic stability, economy and forward progression determinants guiding alternate foot placement were similarly satisfied. Thus the use of virtual planar obstacle in adaptive locomotion study is appropriate. EMG data were used to compute an index relating the changes in muscle activity relative to the normal walking profile. This EMG index was significantly and positively correlated with the amount of foot displacement for the adaptive step. The fact that the dominant choice resulted in minimum foot displacement from its normal landing spot combined with minimal changes in muscle activity validates conclusively the economy determinant. PMID- 16369790 TI - Modulatory effects of high-frequency repetitive transcranial magnetic stimulation on the ipsilateral silent period. AB - In healthy subjects, suprathreshold repetitive transcranial magnetic stimulation (rTMS) at frequencies >2 Hz prolongs the cortical silent period (CSP) over the course of the train. This progressive lengthening probably reflects temporal summation of the inhibitory interneurons in the stimulated primary motor cortex (M1). In this study, we tested whether high-frequency rTMS also modulates the ipsilateral silent period (ISP). In nine normal subjects, suprathreshold 10-pulse rTMS trains were delivered to the right M1 at frequencies of 3, 5, and 10 Hz during maximal isometric contraction of both first dorsal interosseous muscles. At 10 Hz, the second pulse of the train increased the area of the ISP; the other stimuli did not increase it further. During rTMS at 3 and 5 Hz, the ISP remained significantly unchanged. Control experiments showed that 10-Hz rTMS delivered at subthreshold intensity also increased the ISP. rTMS over the hand motor area did not facilitate ISPs in the biceps muscles. Finally, rTMS-induced ISP facilitation did not outlast the 10-Hz rTMS train. These findings suggest that rTMS at a frequency of 10 Hz potentiates the interhemispheric inhibitory mechanisms responsible for the ISP, partly through temporal summation. The distinct changes in the ISP and CSP suggest that rTMS facilitates intrahemispheric and interhemispheric inhibitory phenomena through separate neural mechanisms. The ISP facilitation induced by high-frequency rTMS is a novel, promising tool to investigate pathophysiological abnormal interhemispheric inhibitory transfer in various neurological diseases. PMID- 16369791 TI - Role of force cues in the haptic estimations of a virtual length. AB - This study investigated whether and how the force cues play a role in the haptic perception of length. We assumed that the introduction of a dynamic disruption during haptic exploration generated by a haptic display would lead to a systematic bias in the estimation of a virtual length. Two types of "opposition" disruption ("elastic" and "viscous") were proposed in Experiments 1 and 2, and two types of "traction" disruption ("fluid" and "full") in Experiments 3 and 4. In all experiments, blindfolded adults were asked to compare two lengths of virtual rods explored with the right index. Results revealed an underestimation of the length with elastic and viscous opposition disruptions and an overestimation of this length with fluid and full-traction disruptions. No systematic bias in the estimation was observed in the "control" sessions in which the active exploration of the segment was "normal" (i.e. not disrupted). These results suggest that the forces produced during exploratory movements are used as a relevant cue in the haptic length estimation. PMID- 16369792 TI - Reaching movements in children: accuracy and reaction time development. AB - The present study was undertaken to follow the development of the capability to produce adult-like fast and precise movements reaching visual targets, during childhood. A two-dimensional reaching task was used. We focussed on pre-planning capabilities, by instructing subjects to produce movements as fast as possible, preventing corrections after initiation of movement. The capability of information processing and accurate motor response production were assessed by measuring reaction time (RT, the time elapsed between target presentation and movement onset), movement time (MT, the time elapsed between movement onset and movement end) and precision of response (correlation of response extent and direction with target distance and direction). One child (male) was tested repeatedly since age 6 until age 9. At age 7, RTs decreased. At age 8, accuracy increased, after a temporary decrease at 7. Both accuracy and RT eventually reached the adult level. MTs were similar to the adult ones right from the beginning and they never changed significantly. The results were confirmed in four groups of five children each, aged 6, 7, 8 and 9, respectively. A control group of five adults was also tested. It is concluded that, between age 6 and 9, children become capable of quickly processing visual target information and producing accurate fast and uncorrected reaching trajectories based upon proprioceptive information only, like those typical of adults, by shortening RTs and improving precision, while maintaining adult-like MTs throughout. The capability of quickly reacting to a target acting as a ;Go' signal (measured by RT) and that of information processing to program an accurate motor trajectory (measured by the precision achieved) appear not to be developmentally linked, the former improving earlier, the latter later. PMID- 16369793 TI - Effects of distal and proximal arm muscles fatigue on multi-joint movement organization. AB - To investigate the strategies developed by the central nervous system to compensate for fatigue in muscles, we studied the changes in the relative mechanical contribution of the joint torques in a multi-joint movement following an isometric exhaustion test. Eighteen male subjects performed throws, moving the arm in the horizontal plane, before and after two fatigue protocols. Muscular fatigue was induced either in the distal (extensor digitorum communis) or in the proximal (triceps brachii) agonist muscle of the arm. The kinematic, kinetic and electromyographic parameters of the movement were analysed. The subjects produced two different coordinations following the fatigue protocols. In the distal fatigue condition, the wrist angular velocity was maintained by decreasing elbow active torque. In the proximal fatigue condition, the compensatory strategy involved increasing the contribution of the wrist. In fact, the control of elbow and wrist was modified in order to compensate for the different mechanical effects. PMID- 16369794 TI - Characterization of protein matrix motions in the Rb. sphaeroides photosynthetic reaction center. AB - We use Normal Mode Analysis to investigate motions in the photosynthetic reaction center (RC) protein. We identify the regions involved in concerted fluctuations of the protein matrix and analyze the normalized amplitudes and the directionality of the first few dominant modes. We also seek to quantify the coupling of normal modes to long-range electron transfer (ET). We find that a quasi-continuous spectrum of protein motions rather than one individual mode contributes to light-driven electron transfer. This is consistent with existing theoretical models (e.g. the spin-boson/dispersed polaron model) for the coupling of the protein and solvent "bath" to charge separation events. [Figure: see text]. PMID- 16369795 TI - Learning invariant object recognition in the visual system with continuous transformations. AB - The cerebral cortex utilizes spatiotemporal continuity in the world to help build invariant representations. In vision, these might be representations of objects. The temporal continuity typical of objects has been used in an associative learning rule with a short-term memory trace to help build invariant object representations. In this paper, we show that spatial continuity can also provide a basis for helping a system to self-organize invariant representations. We introduce a new learning paradigm "continuous transformation learning" which operates by mapping spatially similar input patterns to the same postsynaptic neurons in a competitive learning system. As the inputs move through the space of possible continuous transforms (e.g. translation, rotation, etc.), the active synapses are modified onto the set of postsynaptic neurons. Because other transforms of the same stimulus overlap with previously learned exemplars, a common set of postsynaptic neurons is activated by the new transforms, and learning of the new active inputs onto the same postsynaptic neurons is facilitated. We demonstrate that a hierarchical model of cortical processing in the ventral visual system can be trained with continuous transform learning, and highlight differences in the learning of invariant representations to those achieved by trace learning. PMID- 16369796 TI - Modeling electromagnetic fields detectability in a HH-like neuronal system: stochastic resonance and window behavior. AB - Noise has already been shown to play a constructive role in neuronal processing and reliability, according to stochastic resonance (SR). Here another issue is addressed, concerning noise role in the detectability of an exogenous signal, here representing an electromagnetic (EM) field. A Hodgkin-Huxley like neuronal model describing a myelinated nerve fiber is proposed and validated, excited with a suprathreshold stimulation. EM field is introduced as an additive voltage input and its detectability in neuronal response is evaluated in terms of the output signal-to-noise ratio. Noise intensities maximizing spiking activity coherence with the exogenous EM signal are clearly shown, indicating a stochastic resonant behavior, strictly connected to the model frequency sensitivity. In this study SR exhibits a window of occurrence in the values of field frequency and intensity, which is a kind of effect long reported in bioelectromagnetic experimental studies. The spatial distribution of the modeled structure also allows to investigate possible effects on action potentials saltatory propagation, which results to be reliable and robust over the presence of an exogenous EM field and biological noise. The proposed approach can be seen as assessing biophysical bases of medical applications funded on electric and magnetic stimulation where the role of noise as a cooperative factor has recently gained growing attention. PMID- 16369797 TI - [Involuntary patient admission and treatment against patient's will by emergency physicians]. AB - INTRODUCTION: The treatment of acutely ill patients who presumably lack the insight or judgement to determine their need for medical treatment, is a difficult challenge for emergency physicians. We have carried out a study to assess the frequency and relevance of involuntary treatment and procedures in medical emergency services. METHODS: Retrospective chart analysis for a 1-year period was performed for all treatment protocols of a medical emergency service unit and for all court-ordered treatments of a guardianship court. Cases of involuntary treatment by emergency physicians were identified and analysed. RESULTS: In 10.4% of all emergency calls analysed a relevant and acute psychiatric condition was found. In 0.3% of the cases or 4.8% of the psychiatric cases, involuntary inpatient commitment was chosen by the emergency physician. DISCUSSION: Involuntary inpatient commitment by emergency physicians was only necessary in relatively few cases. Nevertheless, in order to be able to correctly consider treatment and management options, emergency physicians should be aware of the basic conditions for treatment without a patient's consent. PMID- 16369798 TI - Should healthy children be vaccinated against influenza? A consensus report of the Summits of Independent European Vaccination Experts. AB - Influenza is often regarded as an illness of the elderly portion of the population because most of the excess mortality associated with influenza epidemics occurs in that age group. However, evidence derived from a large number of clinical studies carried out in different countries and various settings has clearly demonstrated that the burden of influenza is also substantial in children. The attack rates of influenza during annual epidemics are consistently highest in children, and young children are hospitalized for influenza-related illnesses at rates comparable to those for adults with high-risk conditions. Especially among children younger than 3 years of age, influenza frequently predisposes the patient to bacterial complications such as acute otitis media. Children also serve as the main transmitters of influenza in the community. A safe and effective vaccine against influenza has been available for decades, but the vaccine is rarely used even for children with high-risk conditions. Despite several existing problems related to influenza vaccination of children, the current evidence indicates that the advantages of vaccinating young children would clearly outweigh the disadvantages. Considering the total burden of influenza in children, children younger than 3 years of age should be regarded as a high-risk group for influenza, analogously with the age-based definition of high risk among persons 65 years of age or older. Annual influenza vaccination should be recommended to all children from 6 months to 3 years of age. PMID- 16369799 TI - Lethal proliferation of erythroid precursors in a neonate with a germline PTPN11 mutation. AB - We report a neonate with hypertrophic cardiomyopathy and lethal myeloproliferative disorder with excessively proliferating immature erythroid precursors infiltrating non-hematopoietic organs. Mutational analysis uncovered a germline mutation in the Noonan syndrome/LEOPARD syndrome (NS/LS) gene PTPN11. In conclusion, this case report suggests that congenital myeloproliferative disorders in association with germline PTPN11 mutations may affect the erythroid lineage. PMID- 16369800 TI - Nitric oxide accelerates seed germination in warm-season grasses. AB - The nitric oxide (NO) donor sodium nitroprusside (SNP) significantly promoted germination of switchgrass (Panicum virgatum L. cv Kanlow) in the light and in the dark at 25 degrees C, across a broad range of concentrations. SNP also promoted seed germination in two other warm-season grasses. A chemical scavenger of NO inhibited germination and blocked SNP stimulation of seed germination. The phenolic (+)-catechin acted synergistically with SNP and nitrite in promoting seed germination. Acidified nitrite, an alternate NO donor also significantly stimulated seed germination. Interestingly, sodium cyanide, potassium ferricyanide and potassium ferrocyanide at 200 microM strongly enhanced seed germination as well, whereas potassium chloride was without effect. Ferrocyanide and cyanide stimulation of seed germination was blocked by an NO scavenger. Incubation of seeds with a fluorescent NO-specific probe provided evidence for NO production in germinating switchgrass seeds. Abscisic acid (ABA) at 10 microM depressed germination, inhibited root elongation and essentially abolished coleoptile emergence. SNP partially overcame ABA effects on radicle emergence but did not overcome the effects of ABA on coleoptile elongation. Light microscopy indicated extension of the radicle and coleoptiles in seeds maintained on water or on SNP after 2 days. In contrast, there was minimal growth of the radicle and coleoptile in ABA-treated seeds even after 3-4 days. These data indicate that seed germination of warm-season grasses is significantly influenced by NO signaling pathways and document that NO could be an endogenous trigger for release from dormancy in these species. PMID- 16369801 TI - A powerful graphical pulse sequence programming tool for magnetic resonance imaging. AB - A powerful graphical pulse sequence programming tool has been designed for creating magnetic resonance imaging (MRI) applications. It allows rapid development of pulse sequences in graphical mode (allowing for the visualization of sequences), and consists of three modules which include a graphical sequence editor, a parameter management module and a sequence compiler. Its key features are ease to use, flexibility and hardware independence. When graphic elements are combined with a certain text expressions, the graphical pulse sequence programming is as flexible as text-based programming tool. In addition, a hardware-independent design is implemented by using the strategy of two step compilations. To demonstrate the flexibility and the capability of this graphical sequence programming tool, a multi-slice fast spin echo experiment is performed on our home-made 0.3 T permanent magnet MRI system. PMID- 16369803 TI - Development and optimization of a method for analysis of the products from the transesterification of dimethyl carbonate and phenol. AB - A high-purity sample of methyl phenyl carbonate (MPC) was obtained by developing a novel reaction route followed by a series of separation and purification procedures. Identification and quantification of the MPC sample (98.32%) was performed by a gas chromatography-mass spectrometry and Karl Fisher titration. The laboratory-prepared MPC was then used as a standard to optimize quantitative analysis of the products synthesized by transesterification of dimethyl carbonate and phenol. The advantage of the improved method was that MPC can be quantified directly rather than being calculated by subtracting the yield of diphenyl carbonate (DPC) and by-product anisole from the conversion of dimethyl carbonate (DMC). The resulting method was validated for linearity, precision, accuracy, detection limit, and quantification limit. With the improved method, simultaneous accurate quantification of DMC, MPC, DPC, phenol, and anisole in the transesterification products can be achieved. This enables evaluation of the activity and selectivity of different catalysts and control of the reaction processes. PMID- 16369802 TI - Rapid vessel prototyping: vascular modeling using 3t magnetic resonance angiography and rapid prototyping technology. AB - OBJECT: Conversion of thoracic aortic vasculature as measured by Magnetic Resonance Imaging into a real physical replica. MATERIALS AND METHODS: Several procedural steps including data acquisition with contrast enhanced MR Angiography at 3T, data visualization and 3D computer model generation, as well as rapid prototyping were used to construct an in-vitro model of the vessel geometry. RESULTS: A rapid vessel prototyping process was implemented and used to convert complex vascular geometry of the entire thoracic aorta and major branching arteries into a real physical replica with large anatomical coverage and high spatial resolution. CONCLUSION: Rapid vessel prototyping permits the creation of a concrete solid replica of a patient's vascular anatomy. PMID- 16369805 TI - Glow discharge spectroscopy for depth profile analysis: from micrometer to sub nanometer layers. PMID- 16369804 TI - Micellar electrokinetic chromatography-mass spectrometry: combining the supposedly incompatible. PMID- 16369807 TI - Decreased galectin-3 expression during the progression of cervical neoplasia. AB - PURPOSE: Galectin-3 is expressed widely in epithelial and immune cells and the level of expression varies in many cancer cells relative to the normal tissues from which they arise. We investigated whether the expression of galectin-3 is associated with the progression of cervical neoplasia. METHODS: Galectin-3 expression was evaluated in fresh frozen tissues of cervical carcinoma using real time quantitative RT-PCR. In addition, galectin-3 expression was evaluated at the protein level by immunohistochemistry in 90 formalin-fixed paraffin-embedded cervical tissues, 10 normal cervical specimens, 20 low-grade squamous intraepithelial lesions (LSILs), 20 high-grade squamous intraepithelial lesions (HSILs), and 40 invasive squamous cell carcinomas (ISCCs). RESULTS: Real-time quantitative RT-PCR revealed that galectin-3 expressions in tumor cells were significantly downregulated compared with corresponding normal tissue (P=0.005). Moreover, immunohistochemistry showed that galectin-3 expression was strong in all normal cervical squamous epithelia and gradually decreased in accordance with the histopathologic grades in order LSIL>HSIL>ISCC (P<0.001). CONCLUSIONS: These data constitute the first observation that the expression of galectin-3 is downregulated in cervical cancer tissues and suggest that the decreased expression of this galactoside-binding lectin is associated with the progression of cervical neoplasia. PMID- 16369806 TI - The functional analysis of Type I postplasmic/PEM mRNAs in embryos of the ascidian Halocynthia roretzi. AB - Maternal factors, such as a muscle determinant macho-1 mRNA that is localized to the posterior-vegetal cortex (PVC) of fertilized ascidian eggs, are crucial for embryonic axis formation and cell fate specification. Maternal mRNAs that show an identical posterior localization pattern to that of macho-1 in eggs and embryos are called Type I postplasmic/PEM mRNAs. We investigated the functions of five of the nine Type I mRNAs so far known in Halocynthia roretzi: Hr-Wnt-5, Hr-GLUT, Hr PEM3, Hr-PEN1, and Hr-PEN2. Suppression of their functions with specific antisense morpholino oligonucleotides (MOs) had effects on the formation of various tissues: Hr-Wnt-5 on notochord, muscle, and mesenchyme, although zygotic function of Hr-Wnt-5 is responsible for notochord formation; Hr-GLUT on notochord, mesenchyme, and endoderm; and Hr-PEN2 on muscle, mesenchyme, and endoderm. On the other hand, Hr-PEM3 and Hr-PEN1 MOs seemed to have no effect. We conclude that the functions of at least some localized maternal Type I postplasmic/PEM mRNAs are necessary for early embryonic patterning in ascidians. PMID- 16369808 TI - Tissue-type plasminogen activator activity in morphologically normal tissues adjacent to gastrointestinal carcinomas is associated with the degree of tumor progression. AB - PURPOSE: To investigate whether the level of plasminogen activator (PA) activity assayed in gastrointestinal carcinomas and the "morphologically normal tissues" adjacent to them is associated with the degree of tumor progression. METHODS: Tumor and "normal tissues" were obtained from gastrointestinal surgical samples to assess urokinase-type (u-PA) and tissue-type plasminogen activator (t-PA) activities by radial caseinolytic assay and the expression of PA inhibitor-1 (PAI 1) by ELISA. We compared the PA system between the tumor and "normal tissues" and we investigated the existence of correlations between: (a) PA production in the tumor and "normal tissues", (b) different components of the PA system, and (c) PA system and the degree of tumor progression. RESULTS: (1) Total PA activity, u-PA activity and PAI-1 expression are significantly higher in tumor than in "normal tissues", whereas t-PA activity does not differ between them. (2) Total PA activity mainly correlates with u-PA activity in tumor tissues and similarly with u-PA and t-PA activities in "normal tissues". (3) There is a significant association between t-PA activity in tumor and "normal tissues" and the degree of tumor progression. CONCLUSIONS: "Morphologically normal tissues" adjacent to carcinomas present abnormal t-PA activity that is associated with the degree of tumor progression. Assaying of this activity could be useful as a predictive parameter. PMID- 16369810 TI - Posttraumatic bile leaks: role of diagnostic imaging and impact on patient outcome. AB - The aim of this study was to evaluate the impact of bile leaks on patient morbidity and hospital course following blunt and penetrating liver trauma. Forty patients who underwent hepatobiliary scintigraphy after trauma were included. Scintigraphic results were classified as follows: free intraperitoneal bile leak, contained bile leak, and no bile leak. Outcomes measured were length of hospital stay, number of procedures required, and number of subsequent imaging studies. Bile leaks were identified in 25% of patients. Eight percent had free intraperitoneal leaks, 18% contained bile leaks, and 73% had no bile leak. One study was nondiagnostic due to poor hepatic function. Mean hospitalization was as follows: free bile leak group, 53 days; contained bile leak group, 10 days; no bile leak group, 14 days. Patients with free intraperitoneal bile leak had more imaging studies and procedures than patients without free bile leak. Patients with liver injury and free intraperitoneal bile leak have longer hospitalizations and undergo more therapeutic procedures than those without, who respond to conservative management. PMID- 16369811 TI - Short-term effects of hormone therapy on serum C-reactive protein levels in postmenopausal women. AB - OBJECTIVES: The aim of this study was to investigate the effects of tibolone and conjugated equine estrogens (CEEs) plus medroxyprogesterone acetate (MPA) (CEE + MPA) on levels of serum C-reactive protein (CRP), an independent risk factor for cardiovascular disorders, in postmenopausal women. STUDY DESIGN: In this prospective randomized study, we randomly assigned 58 healthy postmenopausal women to CEE (0.625 mg/day) plus MPA (2.5 mg/day) (CEE + MPA) or tibolone (2.5 mg/day). The serum levels of CRP at 3 months after starting treatment were compared with baseline values for both therapies. RESULTS: After 3 months of treatment the median CRP levels increased by 29% in the CEE + MPA group and by 5% in the tibolone group. But, these changes did not have statistical significance (P=0.15, P=0.06, respectively). CONCLUSIONS: Our findings show that neither tibolone nor CEE + MPA caused significant changes in serum CRP levels in postmenopausal women. The potential impact of hormone therapy on serum CRP levels should be investigated in ongoing clinical trials. PMID- 16369812 TI - Macrophage migration inhibition factor is elevated in pregnancy, but not to a greater extent in preeclampsia. AB - BACKGROUND: Maternal serum concentrations of macrophage migration inhibitory factor (MIF) have recently been reported to be elevated in cases with preeclampsia. These findings may be important in increasing our understanding of the underlying events leading to the development of preeclampsia, as this cytokine is also expressed in the placenta, where it has been shown to possess immunemodulatory activities. For this reason we attempted to independently verify this report. METHODS: Plasma levels of MIF were assessed by ELISA in plasma samples collected from normal healthy male and female blood donors (n=20 per group), as well as healthy normal pregnant women in all three trimesters of pregnancy (n=60). In addition, MIF levels were examined from cases with mild and severe preeclampsia (n=20 per study cohort) and matched normotensive pregnancies (n=20). RESULTS: MIF levels were found to be elevated in pregnancy (median=10.1 ng/ml) when compared to non-pregnant controls (median=1.7 ng/ml). A moderate, but not significant, elevation was found to occur from the first to the third trimester of pregnancy. No significant difference was found to occur between the two preeclampsia study groups when compared to the normotensive control group. CONCLUSIONS: Our data suggest that circulatory MIF concentrations are elevated throughout pregnancy, but are not further increased in preeclampsia. PMID- 16369813 TI - Evaluation of the effect of multiple-dose administration of R411, a dual alpha4beta1-alpha4beta7 integrin antagonist on the major CYP isoform activities in healthy volunteers. PMID- 16369814 TI - Similar hormonal responses to concentric and eccentric muscle actions using relative loading. AB - Conventional resistance exercise is performed using sequential concentric (CON) and eccentric (ECC) contractions, utilizing the same muscle load. Thus, relative to maximal CON and ECC resistance, the ECC contraction is loaded to a lesser degree. We have recently shown that at the same absolute load, CON contractions are associated with greater growth hormone (GH) but similar total testosterone (TT) and free testosterone (FT) responses compared with ECC contractions and attributed the larger GH response to greater relative CON loading. In the present study, we have examined the same endocrine parameters to six different upper and lower body exercises using relative loading rather than absolute loading, hypothesizing that GH responses would be similar for CON and ECC actions, but TT and FT responses would be greater after ECC contractions. Seven young men with recreational weight training experience completed an ECC and CON muscle contraction trial on two different occasions in a counterbalanced fashion. The exercises consisted of four sets of 10 repetitions of lat pull-down, leg press, bench press, leg extension, military press, and leg curl exercises at 65% of an ECC or CON 1-RM with 90 s between sets and exercises. CON and ECC actions were performed at the same speed. ECC 1-RMs were considered to be 120% of the CON 1-RM for the same exercise. Blood samples were collected before, immediately after, and 15 min after the exercise. GH significantly increased across both trials but was not different between the two trials. Total testosterone was not significantly altered in response to either trial; however, free testosterone concentrations increased in response to both ECC and CON trials. Data suggest that CON and ECC muscle contractions produce similar GH, T, and free testosterone responses with the same relative loading. PMID- 16369815 TI - Metabolic fate of a large amount of 13C-glycerol ingested during prolonged exercise. AB - We have shown that the oxidation rate of exogenous glycerol and glucose during prolonged exercise were similar when ingested in small amounts (0.36 g/kg) (J Appl Physiol 90:1685,2001). The oxidation rate of exogenous carbohydrate increases with the amount ingested. We, thus, hypothesized that the oxidation rate of exogenous glycerol would also be larger when ingested in large amount. The study was conducted on six male subjects exercising for 120 min at 64 (2)% VO(2)max while ingesting 1 g/kg of (13)C-glycerol. Substrate oxidation was measured using indirect respiratory calorimetry corrected for protein oxidation, and from V(13)CO(2) at the mouth. The (13)C enrichment of plasma glucose was also measured in order to follow the possible conversion of (13)C-glycerol into glucose. In spite of the large amount of glycerol ingested and absorbed (plasma glycerol concentration = 8.0 (0.3) mmol/l at min 100), exogenous glycerol oxidation over the last 80 min of exercise [8.8 (1.6) g providing 4.1 (0.7)% of the energy yield] was similar to that observed when 0.36 g/kg was ingested. The comparison between the (13)C enrichment of plasma glucose and the oxidation rate of (13)C-glycerol showed that a portion of exogenous glycerol was converted into glucose before being oxidized, but also suggested that another portion could have been directly oxidized in peripheral tissues. PMID- 16369816 TI - Intramyocellular lipid and glycogen content are reduced following resistance exercise in untrained healthy males. AB - Resistance exercise has recently been shown to improve whole-body insulin sensitivity in healthy males. Whether this is accompanied by an exercise-induced decline in skeletal muscle glycogen and/or lipid content remains to be established. In the present study, we determined fibre-type-specific changes in skeletal muscle substrate content following a single resistance exercise session. After an overnight fast, eight untrained healthy lean males participated in a approximately 45 min resistance exercise session. Muscle biopsies were collected before, following cessation of exercise, and after 30 and 120 min of post exercise recovery. Subjects remained fasted throughout the test. Conventional light and (immuno)fluorescence microscopy were applied to assess fibre-type specific changes in intramyocellular triacylglycerol (IMTG) and glycogen content. A significant 27+/-7% net decline in IMTG content was observed in the type I muscle fibres (P<0.05), with no net changes in the type IIa and IIx fibres. Muscle glycogen content decreased with 23+/-6, 40+/-7 and 44+/-7% in the type I, IIa and IIx muscle fibres, respectively (P<0.05). Fibre-type-specific changes in intramyocellular lipid and/or glycogen content correlated well with muscle fibre type oxidative capacity. During post-exercise recovery, type I muscle fibre lipid content returned to pre-exercise levels within 120 min. No changes in muscle glycogen content were observed during recovery. We conclude that intramyocellular lipid and glycogen stores are readily used during resistance exercise and this is likely associated with the reported increase in whole-body insulin sensitivity following resistance exercise. PMID- 16369817 TI - Individual differences in the responses to endurance and resistance training. AB - Large individual differences in the responsiveness of cardiorespiratory fitness (VO2peak) to endurance training have been observed in healthy subjects. We tested the hypothesis that subjects with a poor responsiveness to endurance training might benefit from resistance training in terms of aerobic fitness. The study population consisted of sedentary healthy male and female subjects (n=91, 42+/-5 year) assigned to either a training (n=73) or a control group (n=18). The randomized cross-over study design included a 2-week laboratory-controlled endurance or resistance training period with a 2-month detraining period between the interventions. Large individual differences were observed in the changes of VO2peak (DeltaVO2peak) after both the endurance (average 8+/-6 %, P<0.001, range 5 to +22%) and resistance training (average 4+/-5%, P<0.001, range -8 to +16%). The average increase in DeltaVO2peak between genders was similar after both the endurance (8+/-6% for both genders, P=ns) and resistance training (3+/-5% for males and 5+/-6% for females, P=ns). There was no linear relationship between the changes in VO2peak after each training intervention (r=-.09, P=ns). On the contrary, when the study group was divided into quartiles according to the endurance training response (1+/-3, 6+/-1, 9+/-1, and 16+/-3% increase in VO2peak), the group with the lowest response to endurance training increased VO2peak after the resistance training intervention (DeltaVO2peak 7+/-5%, P<0.001). The individual responsiveness of VO2peak to exercise training is related to the mode of training. The healthy males and females whose training response is low after endurance training seem to result in a marked improvement in their cardiorespiratory fitness by resistance training. PMID- 16369818 TI - Immobilization effects in young and older adults. AB - This experiment compared the effects of disuse on the adductor pollicis (AP) muscle in young (YM) and old (OM) men. The AP of the YM and OM was assessed for strength (MVC), compound muscle action potential (CMAP), and volume, and then immobilized for 2 weeks. MVC decreased approximately 22% in the YM, and OM (P<0.001). AP volume was 4.1% (not significant) and 9.5% (P<0.05) less in the YM and OM, respectively. CMAP increased in the OM 0, 24, and 48 h post immobilization, and did not change in the YM. However, the YM showed a greater decrease in specific force as compared to the OM. YM and OM experienced similar losses in strength, yet muscle volume loss was only significant in OM. Although OM are more susceptible to immediate losses in muscle volume, muscle activation strategies appear to preserve strength during atrophy. PMID- 16369819 TI - Effects of pedal frequency on estimated muscle microvascular O2 extraction. AB - An increase in muscle contraction frequency could limit muscle blood flow QM compromising the matching of QM and muscle oxygen uptake VO2M. This study examined the effects of pedal cadence on skeletal muscle oxygenation at low, moderate and peak exercise. Nine healthy subjects [24.7+/-6.3 years (SD)] performed incremental cycling exercise at 60 and 100 rpm. Pulmonary VO2(VO2P) was measured breath-by-breath and vastus lateralis oxygenation was determined by near infrared spectroscopy (NIRS). The deoxyhemoglobin signal ([HHb]) from NIRS was used to estimate microvascular O2 extraction (i.e., [HHb] proportional, variant VO2M/QM). The VO2P and [HHb] for low, moderate and at peak exercise were determined. The VO2P at 60 rpm (low=0.64+/-0.13, moderate=2.03+/-0.38 and peak=3.39+/-0.84 l/min) were lower (P<0.01) than at 100 rpm (1.29+/-0.23, 2.14+/ 0.39 and 3.54+/-0.88 l/min, respectively). There was a progressive increase in [HHb] from low to peak exercise. However, there was no significant difference (ANOVA, P=0.94) for the 60 (in microM, low=24.0+/-9.5, moderate=30.5+/-13.8 and peak=36.7+/-16.5) and 100 contractions/min (in microM, low=25.7+/-11.6, moderate=32.1+/-14.0 and peak=35.4+/-16.5). We conclude that vastus lateralis O2 extraction was similar at 60 and 100 cpm, suggesting that the VO2M/QM in the microcirculation was not altered and, presumably, no impairment of QM occurred with the increase in pedal frequency. PMID- 16369820 TI - Intense exercise induces the degradation of adenine nucleotide and purine nucleotide synthesis via de novo pathway in the rat liver. AB - The purpose of this study was to investigate the influence of intense exercise on the metabolism of adenine nucleotides in the liver. In the first experiment, to determine the degradation of adenine nucleotides, hepatic adenine nucleotides of rats were labeled by an intraperitoneal administration of 15N-labeled adenine the day before treadmill running to exhaustion. In the second experiment, to determine the de novo synthesis of purine nucleotides after intense exercise, 14C glycine was intraperitoneally administered to rats performing intense running on a treadmill. In the first experiment, hepatic levels of ATP and total adenine nucleotides showed a reduction immediately after exercise. In contrast, hepatic levels of AMP, adenosine, hypoxanthine and uric acid showed an increase immediately after exercise. The hepatic 15N level continued to decline during the recovery period after exercise. Urinary excretion of 15N-urate was 40% higher in the exercised rats than in the control rats. In the second experiment, the radioactivity of 14C detected in the fraction of hepatic urate and allantoin was approximately 300% higher in the exercised rats than in the control rats. 14C radioactivity that excreted into urine as urate and allantoin was approximately 200% higher in the exercised rats. Intense exercise led to the degradation of hepatic adenine nucleotides, which were not utilized for the re-synthesis of nucleotide and further degraded to hypoxanthine or uric acid. Intense exercise induced the synthesis of purine nucleotides in the liver via a de novo pathway and these synthesized nucleotides were also degraded to nucleosides and excreted into urine. PMID- 16369821 TI - Pharmacokinetics and pharmacodynamics of irinotecan and its metabolites from plasma and saliva data in patients with metastatic digestive cancer receiving Folfiri regimen. AB - PURPOSE: Irinotecan is extensively metabolized into at least four compounds and previous pharmacokinetic-pharmacodynamic studies have given varying results. We hypothesized that saliva, a noninvasive, safe and painless biological sampling process, could be a good predictor of the behavior of irinotecan and its metabolites. METHODS: Thirty-five patients with metastatic digestive cancer were treated with a Folfiri regimen every 2 weeks. The irinotecan-administered dose was 180 mg/m(2); 17 patients participated in a dose-escalating study. Irinotecan and its metabolites (SN-38, SN-38G, APC, NPC) were quantified in plasma and saliva by high-performance liquid chromatography with fluorescence detection. RESULTS: The mean irinotecan systemic clearance and steady-state volume of distribution values were 14.3 l/h/m(2) and 211 l/m(2), respectively. The intrapatient variability (22-28%) was far lower than the interindividual variability (33-88%). Age and weight were the two physiological parameters that influenced drug disposition. For irinotecan, SN-38, APC and NPC, similar pharmacokinetic profiles were observed from plasma and saliva data. The saliva/plasma AUC ratios averaged 1 for irinotecan, 0.3 for SN-38, 0.17 for APC and 0.27 for NPC. Neutropenia, diarrhea and nausea were the main toxicities encountered. From both plasma and saliva data, the percentage decrease in neutrophil count appeared to be related to irinotecan and SN-38 AUCs. CONCLUSIONS: All these findings provide a rationale for an individual adaptation of irinotecan dosing. In case of difficult venous access, the titration of irinotecan and of its active metabolite SN-38 in saliva may prove relevant. PMID- 16369822 TI - Expression of IGFBPs in the developing mouse submandibular and von Ebner's glands. AB - The expression of insulin-like growth factor binding proteins (IGFBPs) in the developing mouse submandibular and von Ebner's glands was determined by in situ hybridization and by an immunohistochemical method. In the submandibular glands, IGFBP-2 and IGFBP-4 mRNAs were expressed in the terminal end-buds (TEB) at E13 E17, concomitant with epithelial branching. IGFBP-3 mRNA was expressed in the mesenchyme surrounding the TEB; and IGFBP-5 mRNA, in the ducts. At E17, IGFBP-5 mRNA expression was observed not only in the ducts but also in the TEB. Similarly, IGFBP-4 mRNA expression was observed not only in the TEB but also in the mesenchyme. After birth, IGFBP-4 expression was observed only in the connective tissue and disappeared by P14. That of IGFBP-7 appeared at P1 and was observed in the connective tissue until P21. The IGFBP-5 mRNA expression pattern after birth was the same as that seen at E17, but at P21 IGFBP-5 was immunohistochemically expressed only in the duct. The mRNA level of IGFBP-2 expression at postnatal days was weak, but its protein was detected in the ducts and acini at P14-P21. In von Ebner's glands, which appeared at the base of the circumvallate papillae at E17, only IGFBP-2 and IGFBP-4 mRNAs were expressed in the ducts and acini. Postnatally, IGFBP-4 was substituted by IGFBP-5 in the same region. Immunohistochemically, IGFBP-5 and IGFBP-2 were expressed in the ducts and acini at P14-P21. Throughout the study, IGFBP-6 was not detected by in situ hybridization, the immunoreactivity for it was observed in the nerve fibers of submandibular and von Ebner's glands. These data support a role for these molecules as local mediators of salivary growth and differentiation. PMID- 16369823 TI - Role of Wnt-6 in limb myogenesis. AB - Cells from the ventrolateral lip of the dermomyotome at limb levels undergo epithelio-mesenchymal transition and migrate as individual and undifferentiated cells into the limb buds. The precursor cells are under the influence of various signaling factors in the limb. Dorsal and ventral ectoderm influences various aspects of limb development. In addition to our previous studies, we investigated the influence of ectoderm and Wnt-6 on somitic cells in the limb bud. We show that in the absence of ectoderm the precursor cells never form muscle cells but differentiate into endothelial cells. In addition, we show that Wnt-6 that is secreted from the ectoderm influences the precursor cells to form muscle even in the absence of ectoderm. This indicates that Wnt-6 is an ectodermal signal that induces somite-derived progenitor cells to form muscle cells during limb development. PMID- 16369824 TI - [Pulmonary tumors]. AB - Pulmonary nodules may be caused by true neoplasms of the airways, respiratory epithelium, vessels and connective tissue which are mainly malignant or metastatic. They may, however, also represent mainly inflammatory benign pseudotumours. Non-small cell lung cancer (NSCLC) usually presents as an ill defined soft-tissue pulmonary nodule or mass, often without lymphadenopathy; endobronchial lesions are less common. Small cell lung cancer (SCLC) commonly presents with extensive lymph node and hematogenous metastases. Carcinoid tumors typically manifest as endobronchial lesions with secondary obstruction. Sarcomas are rare and show variable morphology. Metastases most commonly present as basal, peripheral, well-defined nodules. Granulomas often exhibit typical calcifications; hamartoma may also contain popcorn-type calcifications as well as fat. Pulmonary arteriovenous malformation is characterised by feeding artery and draining vein. Inflammatory nodules are well vascularized and may regress spontaneously; their differentiation from malignant lesions, however, is usually difficult. PMID- 16369825 TI - [Fetal magnetic resonance imaging. Diagnostics in cases of congenital cystadenomatoid malformation of the lung (CCAM)]. AB - Despite advancing therapeutic strategies, congenital cystadenomatoid malformation of the fetal lung is still a potentially life-threatening anomaly. Antenatally, the development of hydrops as well as the natural history of the malformation is of particular therapeutic and prognostic importance. Postnatally, respiratory distress due to pulmonary hypoplasia counts as a crucial limiting factor. Owing to its feasibility and practicability, as well as a high sensitivity, antenatal ultrasound is still the screening method of choice for the detection of fetal thoracic malformations. However, particularly in cases of indistinguishable sonographic findings, fetal MRI is the modality of choice for proving the diagnosis and preliminary appraisal of intensive care therapy and extracorporal membrane oxygenation postnatally. Furthermore, fetal MRI often facilitates assessment and planning of intrauterine surgical procedures. These two features frequently require a close transfer to an expert neonatal centre. PMID- 16369826 TI - Correlation between respiration-induced thoracic expansion and a shift of central structures. AB - The fusion of computed tomography (CT) and positron emission tomography (PET) may improve diagnostic accuracy, but is limited by different breathing protocols. This study aimed at quantifying respiration-induced alignment errors. PET-CT was acquired in 24 patients. Contrast-enhanced whole-body CT was obtained in a single breath hold in the expiratory state of a normal breathing cycle. An inspiratory low-dose CT of the thorax was acquired in the same session, and comparison of the two CT scans was used to assess the potential mismatch of PET and CT fusion. The largest craniocaudal expansion was found in the area of the diaphragm. A considerable sagittal expansion was found in the anterior parts of the lungs. Central tracheo-bronchial structures were displaced during inspiration mainly in the anterior and caudal directions. The craniocaudal shift of central structures showed a linear correlation with the diaphragmatic expansion, whereas the sagittal shift correlated with the sagittal pleural expansion. There was, however, no correlation between craniocaudal and sagittal respiratory motion. Alignment errors are most severe in the base of the lung, but central structures are affected, too. Understanding of the main vectors of respiratory motion may help in image interpretation when PET and CT are acquired separately. PMID- 16369827 TI - Patient-based outcomes for the operative treatment of degenerative lumbar spinal stenosis. AB - STUDY DESIGN: A retrospective surgical case series was conducted using a condition-specific, patient-based outcomes assessment. OBJECTIVE: The goal of this study was to describe the outcome predictors of decompressive surgery for degenerative lumbar spinal stenosis (DLSS). SUMMARY OF BACKGROUND DATA: Surgical decompression is the recommended treatment for patients with moderate to severe DLSS. Previous studies have reported that factors such as the number of operated levels and patient health status are predictors of surgical outcomes. METHODS: This study analyzed the success rates of 58 DLSS patients treated with decompressive surgery. Outcomes were measured with the Zurich Claudication Questionnaire (ZCQ) completed pre-operatively and at least 12 months post operatively (range 12-54 months). The ZCQ includes three distinct domains that involve symptom severity, physical function, and patient satisfaction. Variables such as age, sex, pre-operative symptom severity, and arthrodesis were analyzed as predictors of success. RESULTS: The study group included 21 males and 37 females, and the mean age of all patients was 66 years (range 41-80 years). Overall, 63.8% of the patients had significant clinical improvement in Symptom Severity, 55.2% had significant clinical improvement in Physical Function, and 58.6% of the patients were at least somewhat satisfied; 63.8% (37/58) of the patients were considered to be clinically successful. Patients with more severe pre-operative symptoms and more physical function restrictions had better success results than those patients with milder symptoms and less restrictive physical function. Also, patients who were followed for less than 24 months had better success than those followed for more than 24 months. There was no significant difference in the clinical success rates of (1) patients who were fused and those not fused, (2) males and females, (3) patients aged less than 65 years and those greater than 65 years, and (4) patients who were treated at one or two levels and those treated at three or four levels. CONCLUSION: The results of this retrospective study indicate that operative decompression of the lumbar spine offers significant improvement for patients with DLSS. Although not all comparisons were statistically significant, there was a trend for DLLS patients aged less than 65 years with more severe pre-operative symptoms and physical function disturbances treated at one or two levels with a laminectomy and fusion to have the best outcomes. PMID- 16369828 TI - Health economic evaluation in lumbar spinal fusion: a systematic literature review anno 2005. AB - The goal of this systematic literature review was to assess the evidence for cost effectiveness of various surgical techniques in lumbar spinal fusion in conformity with the guidelines provided by the Cochrane Back Review Group. As new technology continuously emerges and divergent directions in clinical practice are present, economic evaluation is needed in order to facilitate the decision makers' budget allocations. NHS Economic Evaluation Database, MEDLINE, EMBASE and Cochrane Library were searched. Two independent reviewers (one clinical content expert and one economic content expert) applied the eligibility criteria. A list of criteria for methodological quality assessment was established by merging the criteria recommended by leading health economists with the criteria recommended by the Cochrane Back Review Group. The two reviewers independently scored the selected literature and the disagreement was resolved by means of consensus following discussion. Key data were extracted and the level of evidence concluded. Seven studies were eligible; these studies reflected the diversified choices of economic methodology, study populations (diagnosis), outcome measures and comparators. At the conclusion of quality assessment, the methodological quality of three studies was judged credible. Two studies investigated posteolateral fusion (PLF) +/- instrumentation in different populations: one investigated non-specific low back pain and one investigated degenerative stenosis + spondylolisthesis. Both studies reflected that cost-effectiveness of instrumentation in PLF is not convincing. The third study concerned the question of circumferential vs anterior lumbar interbody fusion and found a non significant difference between the techniques. In conclusion, the literature is limited and, in view of the fact that the clinical effects are statistically synonymous, it does not support the use of high-cost techniques. There is a great potential for improvement of methodological quality in economic evaluations of lumbar spinal fusion and further research is imperative. PMID- 16369829 TI - Comments on "In vivo demonstration of the effectiveness of thoracoscopic anterior release using the fulcrum bending radiograph: a report of five cases" (Cheung et al.). PMID- 16369830 TI - Outcome scores in spinal surgery quantified: excellent, good, fair and poor in terms of patient-completed tools. AB - Outcome scores are very useful tools in the field of spinal surgery as they allow us to assess a patient's progress and the effect of various treatments. The clinical importance of a score change is not so clear. Although previous studies have looked at the minimum clinically important score change, the degree of score change varies considerably. Our study is a prospective cohort study of 193 patients undergoing discectomy, decompression and fusion procedures with minimum 2-year follow-up. We have used three standard outcome measures in common usage, the oswestry disability index (ODI), the low back outcome score (LBOS) and the visual analogue score (VAS). We have defined each of these scores according to a global measure of outcome graded by the patient as excellent, good, fair or poor. We have also graded patient perception and classified excellent and good as success and fair and poor as failure. Our results suggest that a median 24-point change in the ODI equates with a good outcome or is the minimum change needed for success. We have also found that different surgical disorders have very different minimal clinically important differences as perceived by patient perception. We found that for a discectomy a minimum 27-point change in the ODI would be classed as a success, for a decompression the change in ODI needed to class it as a success would be 16 points, whereas for a fusion the change in the ODI would be only 13 points. We believe that patient-rated global measures of outcome are of value and we have quantified them in terms of the standard outcome measures used in spinal surgery. PMID- 16369831 TI - In-vivo demonstration of the effectiveness of thoracoscopic anterior release using the fulcrum-bending radiograph: a report of five cases. AB - Thoracoscopic anterior release of stiff scoliotic curves is favored because of its minimally invasive nature. Animal and human cadaveric studies have shown that it can effectively improve spinal flexibility in non-scoliotic spines; however it has not been demonstrated to be effective in actual patients with scoliosis. The fulcrum-bending radiograph has been shown to accurately reflect the post operative correction. To demonstrate that the flexibility was increased after the anterior release; five patients with idiopathic thoracic scoliosis who underwent staged anterior thoracoscopic release and posterior spinal fusion were assessed using the fulcrum-bending radiograph. The average number of discs excised was four. Spinal flexibility as revealed by the fulcrum-bending technique, was compared before and after the anterior release. The patients were followed for an average of 4 years (range 2.2-4.9 years). Fulcrum-bending flexibility was increased from 39% before the thoracoscopic anterior spinal release to 54% after the release (P<0.05). The average Cobb angle before the anterior release was 71 degrees on the standing radiograph and 43 degrees with the fulcrum-bending radiograph. This reduced to 33 degrees on the fulcrum-bending radiograph after the release, and highly corresponded to the 30 degrees measured at the post operative standing radiograph and at the latest follow-up. Previous animal and cadaveric studies demonstrating the effectiveness of thoracoscopic anterior release did not have scoliosis. We are able to demonstrate in patients with adolescent idiopathic scoliosis, that thoracoscopic anterior spinal release effectively improves the spinal flexibility. PMID- 16369832 TI - Effectiveness of multiple-level decompression in laminoplasty and simultaneous C1 laminectomy for patients with cervical myelopathy. AB - A retrospective study to investigate the relationship between the surgical levels and decompression effects was performed in patients with cervical myelopathy who had undergone Tension-band laminoplasty (TBL) with/without simultaneous C1 laminectomy. One hundred and sixty-eight patients (115 males, 53 females; age: 31 80 years, average 58.9 years; follow-up period: 12-120 months, average 20 months) were divided into three groups according to the range of the surgical levels: seventy-two patients in group A underwent TBL at the C2-C7 levels with C1 laminectomy; 60 patients in group B underwent TBL at the C2-C7 levels; 36 patients in group C underwent TBL at the C3-C7 levels. Neurological evaluation was performed by using the Japanese Orthopedic Association (JOA) scoring system. The alignment changes of the spinal column and the spinal cord were analyzed using pre- and post-operative roentgenograms and MRIs. The differences in the pre and post-operative anterior subarachnoid spaces (D-ASAS), the spinal cord diameters (D-CORD), and the dural sleeve diameters (D-DURA) at the C1-C7 levels were also analyzed by using MRIs. The JOA scores improved in all groups. As for the spinal alignment, neither significant changes between pre- and post-operation in any group nor significant differences among the three groups were found. The lordosis of the cervical spinal cord was decreased in all groups. D-ASAS of group A was larger than that of group B at the C1-C5 levels (P<0.05), as were those of D-CORD and D-DURA at the C1-C2 and C4-C5 levels (P<0.05). D-ASAS of group A was larger than that of group C at the C1-C4 levels (P<0.05), as were those of D-CORD and D-DURA at the C1-C5 levels (P<0.05). In conclusion, laminoplasty including the C2-C7 levels with simultaneous C1 laminectomy was proven to allow the most posterior shift of the spinal cord within the widened dural sleeve at C5 or higher levels without significantly changing the spinal alignment. PMID- 16369833 TI - AO-classification of thoracic and lumbar fractures--reproducibility utilizing radiographs and clinical information. AB - This study was designed to assess the inter-observer reliability and intra observer reproducibility of standard radiographic evaluation of 150 thoraco lumbar fractures using the AO-classification. The influence of clinical information on agreement levels was also evaluated. Six observers (two junior and four senior residents) evaluated the radiographic images. The injuries were classified by each observer as either type A, B or C according to the AO classification system and the levels of agreement were documented. After 3 months the injuries were again classified with the addition of the clinical findings of each patient and the level of agreement evaluated. The level of agreement was measured using Cohen's kappa-test. The overall inter-observer agreement was rated as fair (0.291) in the first session and moderate (0.403) in the second. Intra observer values ranged from slight (0.181) to moderate (0.488). The increased level of agreement in the second session was attributed to the value of additional clinical information, the learning curve of the junior residents and the simplicity of the classification. PMID- 16369834 TI - Effect of serotonin depletion on 5-HT2A-mediated learning in the rabbit: evidence for constitutive activity of the 5-HT2A receptor in vivo. AB - RATIONALE: Associative learning during Pavlovian eyeblink conditioning has been shown to be regulated by 5-HT2A receptors. The existence of inverse agonists that retard learning through an action at the 5-HT2A receptor suggests the existence of constitutive activity at that receptor and that depletion of serotonin should have minimal effects on learning. OBJECTIVES: We examined whether depletion of serotonin would impair trace eyeblink conditioning or the enhancement of conditioning produced by the agonist lysergic acid diethylamide (LSD) and the retardation of conditioning produced by the inverse agonist MDL11,939. METHODS: Animals received bilateral intraventricular injections of 5,7-dihydroxytryptamine (5,7-DHT) at doses of 760 or 1,140 microg/side (1.88 or 2.82 micromol/side) and were later exposed to eight daily conditioning sessions. RESULTS: Serotonin depletion produced by the lower dose of 5,7-DHT was 71 and 72% in cortex and hippocampus, respectively, with no change in 5-HT2A receptor density, no effect on learning, and no effect on the ability of LSD to enhance and MDL11,939 to retard learning. The higher dose of 5,7-DHT produced serotonin decreases of 85 and 90% in cortex and hippocampus, respectively, accompanied by a 96% decrease in the density of the serotonin transporter, but no significant effect on learning. CONCLUSIONS: Pavlovian trace eyeblink conditioning is regulated predominantly by the constitutive activity of the 5-HT2A receptor rather than by serotonin release onto the receptor during learning. It was suggested that the 5-HT2A receptor regulates learning by modulating the release of dopamine, acetylcholine, and glutamate, transmitters known to affect eyeblink conditioning. PMID- 16369835 TI - Cocaine self-administration under fixed and progressive ratio schedules of reinforcement: comparison of C57BL/6J, 129X1/SvJ, and 129S6/SvEvTac inbred mice. AB - RATIONALE: Combining strains to generate mutant mice may obscure conclusions regarding the targeted gene. Specifically, cocaine may have reduced reinforcing effects in 129 substrains compared to the C57BL/6 strain, commonly used for ES cells and breeding, respectively. OBJECTIVES: We tested the hypothesis that reinforcing effects of cocaine differ between the C57BL/6J strain and two substrains of 129, 129X1/SvJ and 129S6/SvEvTac. METHODS: To assess and reduce performance differences, operant responding was established with liquid food as a reinforcer and evaluated under fixed and progressive ratio schedules. Dose-effect functions for intravenous cocaine self-administration were then determined under both schedules. Finally, reinforced and nonreinforced manipulanda were reversed to assess acquisition of self-administration using a previously nonreinforced response. RESULTS: Relative to C57BL/6J mice, 129X1/SvJ mice showed decreased reinforcing effects of low-magnitude food and cocaine reinforcers. Dose-effect functions for cocaine self-administration were comparable between C57BL/6J and 129S6/SvEvTac mice, despite delayed acquisition of operant behaviors and rightward shifts in the food concentration-effect functions in 129S6/SvEvTac mice. A high cocaine dose clearly served as a positive reinforcer in all three strains in a reversal procedure. CONCLUSIONS: Relative to C57BL/6J mice, the reinforcing effects of cocaine were diminished in 129X1/SvJ mice, but only for low cocaine doses, and a similar profile was observed with food reinforcement. 129S6/SvEvTac mice required more extensive operant training than C57BL/6J mice did, but after acquisition, reinforcing effects of cocaine were similar in the two strains. We suggest that comparable phenotypes observed in gene-targeting studies may result from genetic background, whereas more profound or qualitatively different phenotypes may be more confidently attributed to targeted mutations. PMID- 16369837 TI - PET and CT demonstration of metastatic invasion of the thoracic duct. PMID- 16369836 TI - Prefrontal and temporal gray matter density decreases in opiate dependence. AB - RATIONALE: There have been only a few structural brain-imaging studies, with varied findings, of opiate-dependent subjects. Voxel-based morphometry (VBM) is suitable for studying whole brain-wise structural brain changes in opiate dependent subjects. OBJECTIVES: The objective of the current study is to explore gray matter density in opiate-dependent subjects. METHODS: Gray matter density in 63 opiate-dependent subjects and 46 age- and sex-matched healthy comparison subjects was compared using VBM. RESULTS: Relative to healthy comparison subjects, opiate-dependent subjects exhibited decreased gray matter density in bilateral prefrontal cortex [Brodmann areas (BA) 8, 9, 10, 11, and 47], bilateral insula (BA 13), bilateral superior temporal cortex (BA 21 and 38), left fusiform cortex (BA 37), and right uncus (BA 28). CONCLUSIONS: This study reports that opiate-dependent subjects have gray matter density decreases in prefrontal and temporal cortex, which may be associated with behavioral and neuropsychological dysfunction in opiate-dependent subjects. PMID- 16369838 TI - Technetium-99m red blood cell imaging of multicentric kaposiform haemangioendothelioma. PMID- 16369839 TI - The S18Y polymorphism in the UCHL1 gene is a genetic modifier in Huntington's disease. AB - An expanded polyglutamine stretch in the huntingtin protein has been identified as the pathogenetic cause of Huntington's disease (HD). Although the length of the expanded polyglutamine repeat is inversely correlated with the age-at-onset, additional genetic factors are thought to modify the variance in the disease onset. As linkage analysis suggested a modifier locus on chromosome 4p, we investigated the functional relevance of S18Y polymorphism of the ubiquitin carboxy-terminal hydrolase L1 in 946 Caucasian HD patients. In this group, the allelic variation on locus S18Y is responsible for 1.1% of the variance in the HD age-at-onset, and the rare Y allele is associated with younger-aged cases. PMID- 16369840 TI - Genetic transformation of Ascochyta rabiei using Agrobacterium-mediated transformation. AB - In order to study pathogenic mechanisms of the plant pathogen Ascochyta rabiei, conditions for efficient transformation using Agrobacterium-mediated transformation were investigated. Hygromycin B resistance (hph) was superior to geneticin resistance (nptII) for selecting transformants, and the hph gene was more efficiently expressed by the Aspergillus nidulans trpC promoter than by the Cauliflower mosaic virus 35S promoter CaMV35S. Co-cultivation on solid media for 72 h was optimal for generating transformants, but increasing the ratio of bacterial cells to conidia did not affect transformation efficiency. All hygromycin B-resistant transformants carried transfer-DNA (T-DNA) as determined by polymerase chain reaction (PCR) and the T-DNA integrations appeared to be random and in single copy as detected by Southern hybridization. Transformants remained resistant to hygromycin B in the absence of selection. Variations in colony morphology were observed in the presence of hygromycin B under different culture conditions, and a variety of altered phenotypes including reduced virulence were observed among 550 transformants. Inverse PCR was more efficient than TAIL-PCR in identifying flanking genomic sequences from T-DNA borders, and the possible causes are discussed. This transformation technique and recovery of flanking DNA using inverse PCR will provide a useful tool for genetic studies of A. rabiei. PMID- 16369841 TI - Case-control analysis of the financial cost of medication errors in hospitalized patients. AB - The cost of medication errors (MEs), and by extension their impact on the running of hospitals, is generally unknown to hospital managers. This study estimated the financial costs involved in the additional use of hospital resources as a result of MEs. For this we used a database covering 20,014 hospital admissions, with clinical and personal information about each patient, the costs related to his/her stay, and physicians' prescriptions. Analyses were carried out using cases and control techniques to calculate the additional cost of MEs. During the study period, 2001, the analysis indicated that the MEs analyzed caused an additional 303 days of hospital stay, with an overall annual cost of nearly 6,000 euros. Our study confirms that MEs are a costly reality. The presence of MEs doubled the cost per patient. PMID- 16369842 TI - Acute tubulointerstitial nephritis: possible association with cytomegalovirus infection. PMID- 16369843 TI - Brachydactyly short-stature hypertension syndrome: a case with associated vascular malformations. AB - Brachydactyly short-stature hypertension syndrome, also known as hypertension with-brachydactyly (HTNB) syndrome, is a rare autosomal dominant disorder that was first described by Bilginturan and colleagues in 1973. Many familial cases of HTNB have been reported, but the first sporadic case of this condition was published only recently. This article describes a case of HTNB syndrome in a 16 year-old boy. Although Doppler ultrasonography of the kidneys and renal arteries showed normal findings, magnetic resonance angiography showed an aberrant right posterior inferior cerebellar artery, early bifurcation of the left renal artery, and irregularity and stenosis of the inferior dominant branch of this artery. The patient's father was in chronic renal failure of which the primary pathology was unknown. We speculate that the described case is the second documented sporadic case of HTNB syndrome. This disorder should be included in the differential diagnosis of patients with short stature and hypertension of unknown aetiology. Such individuals should be carefully examined for brachydactyly and for cerebral cerebellar and renal vascular malformations. PMID- 16369844 TI - Childhood membranous nephropathy in association with interstitial nephritis and Fanconi syndrome. PMID- 16369845 TI - [Wolfgang Amadeus Mozart. A urological pathography]. AB - The death of Wolfgang Amadeus Mozart was mysterious from the very first day, and cause of wildest speculation and adventurous assertions. Over the last 100 years, medical science has investigated the physical sufferings and the mysterious death of Mozart with increasing intensity. By means of letters from his father Leopold, his sister "Nannerl", himself and reports from his physicians and contemporaries, we would like to create a medical pathography. The rumour that Mozart was poisoned appeared soon after his early death at the age of 35 on December 5th 1791, and was kept up persistently. Accused were the physician van Swieten, Mozart's freemason's loge and the royal band master Salieri. Mozart, however, died due to chronic kidney disease and ultimately due to uraemia. Once the renal damage has reached a certain point, a minimum of additional stress leads to decompensation. This catastrophe occurs typically within the fourth decade of life. When listening to Mozart's music, we should remember that this apparently happy person was actually a premature adult robbed of his childhood, whose short life was an endless chain of indisposition, over fatigue, misery, concern and illness. PMID- 16369846 TI - [Revision of tumour endoprostheses around the knee joint. Review and own results]. AB - Revision of tumour endoprostheses around the knee joint is mainly caused by aseptic loosening and deep infection. Mechanical complications have been significantly reduced by improvements in prosthetic design. The rate of aseptic loosening after distal femur or proximal tibia replacement ranges from 10% to 45% depending on the time of follow-up and prosthetic design; the success rate after revision averages 75% at midterm follow-up. The rate of infection reported in the literature varies between 2.9% and 12%. One stage revision may be considered after acute infection without signs of loosening, after which a success rate of 77.8% can be expected. Two stage revision is mainly indicated in multi-resistant microorganisms as well as septic loosening, with a success rate of 75% after 5 years. Deep infection represents the most serious complication in long-term follow-up, often leading to amputation due to soft tissue problems after tumour resection. The authors were able to confirm the minimal decrease in MSTS score despite the large number of operations (15 patients: nine females, six males; median MSTS score 84%; mean 81%). PMID- 16369847 TI - [Revision of unicompartmental knee arthroplasty]. AB - Despite the good midterm survivorship reported for unicondylar knee arthroplasty, an increase in revision surgery has to be expected due to increased replacement rates. The reasons for failure as well as distribution are different for unicondylar knee arthroplasty compared to total knee arthroplasty. The main reasons for revision are aseptic loosening and the progression of osteoarthritis. In most cases, unicondylar knee arthroplasty will be revised to total knee arthroplasty. To obtain good revision results, the cause of implant failure has to be analysed carefully. In the case of contained bone defects, the reconstruction can be supported with bone grafting. For those cases with uncontained defects, implants with augmentation and, in some cases, stem extensions are needed. The modularity of the revision implant should cover different intraoperative requirements. PMID- 16369848 TI - Sudden death after isobutane sniffing: a report of two forensic cases. AB - The intentional inhalation of a volatile substance ("sniffing") causing euphoria and hallucinations is an under-recognised form of substance abuse in children and adolescents with a high morbidity and mortality. Sudden death can be caused by cardiac arrhythmia, asphyxia or trauma. Two fatal cases of isobutane sniffing of cigarette lighter refill containing isobutane are reported. Toxicological investigations revealed the presence of isobutane in the heart blood and brain tissue of both cases (case 1: heart blood 0.1 microg/g, brain tissue 2.3 microg/g; case 2: heart blood 4.6 microg/g, brain tissue 17.4 microg/g) and the presence of its metabolite 2-methyl-2-propanol in the heart blood of both cases (0.5 and 1.8 microg/g, respectively). The histological investigations of the inner organs showed similar results in both victims. Autopsy findings, results of the histological and immunohistochemical investigations, toxicological findings and analytical procedures are discussed. PMID- 16369849 TI - The symbiotic fauna of the African termite Hodotermes mossambicus identification of four flagellate species of the genera Spironympha, Trichomonoides and Retortamonas. AB - Two Spironympha species were described by light immunofluorescence and electron microscopy. Spironympha fibrosa n. sp. has a narrow columella, several axostylar fibres and original striated myoneme-like fibres attached to the last basal body of each flagellar line. Spironympha simplex n. sp. is small and has non-striated microfibrils attached to the basal body section and an axostyle composed of two fibres. Trichomonas termitidis, as reported by Dogiel (Researches on parasitic protozoa from the intestine of termites. II. Lophomonadidae. Sci Res Zool Exped Brit E Africa made by Prof. V. Dogiel and I. Sokolow in 1914 10:20-35, 1917), has the features of Trichomonoides trypanoides, as reported by Brugerolle and Bordereau (Eur J Protistol 40:163-174, 2004). Retortamonas hodotermitis n. sp. shares the characteristics of insect retortamonads, as reported by Brugerolle (Protistologica 8:233-240, 1976), but does not correspond to any species described to date. The species identification differs from that of described in Dogiel (Russkii Arkhiv Protistologii 1:172-234, 1922). PMID- 16369850 TI - Mature neuroblastic tumors with spinal cord compression: report of five pediatric cases. AB - BACKGROUND: Neuroblastic tumors cause spinal cord compression when they arise primarily in the spinal canal or invade it through the radicular foramen. Whereas neuroblastomas (NB) are relatively common and are generally treated with chemotherapy, mature neuroblastic tumors (MNT), which include intermixed ganglioneuroblastomas (iGNB) and ganglioneuromas (GN), are less common and the role of surgery is more prominent. Because MNT are rare and have been separated only recently from NB, their clinical and radiological features as well as the role of surgery are poorly defined. MATERIALS AND METHODS: In order to increase our knowledge on MNT, we reviewed our database for cases operated for spinal cord compression in our department since the introduction of magnetic resonance imaging (MRI). We treated four cases of NB and one case of iGNB presenting primarily with spinal cord compression. REPORT OF CASES: MNT represented 10% of spinal tumors and 1.6% of all tumors of the nervous system in our pediatric neurosurgical practice. The neurological and oncological outcomes were generally favorable after surgical resection, followed by orthotic treatment. In one case with neurofibromatosis type 1, the tumor was inoperable and the child died of tumor progression several years later. CONCLUSION: GN, and some iGNB, are chemo insensitive and can only be cured by surgical removal. Surgery is an emergency in case of rapidly progressing paraplegia and can be challenging because the tumor is often hard and hemorrhagic. In case of subtotal removal, tumor remnants can stay stable without oncological treatment. PMID- 16369851 TI - Non-anaplastic pleomorphic xanthoastrocytoma with neuroradiological evidences of leptomeningeal dissemination. AB - CASE REPORT: A case of a non-anaplastic pleomorphic xanthoastrocytoma (PXA) presenting with leptomeningeal dissemination (LMD) affecting a 9-year-old girl is presented. DISCUSSION: The neoplasia in this young girl had the otherwise classical clinical features of PXA: the relatively advanced paediatric age of the patient, the seizures as presenting sign; the primary site in the temporal lobe; and the MRI findings of the partially solid and cystic superficial lesion. Only the tumour involvement of the chiasma and the infundibulus was a relatively unusual finding. In a 5-year period, the tumour underwent malignant transformation, bringing the child to death because of the primary tumour progression. However, the leptomeningeal deposits remained unchanged throughout the clinical course. CONCLUSION: To our knowledge, this is the first case of a non-anaplastic PXA presenting with disseminated disease. Thus, it was thought important to describe this case in order to add further information regarding the spectrum of the presenting clinical features of this rare neoplasm and the phenomenon of LMD of non-malignant glioma. PMID- 16369852 TI - Surgical treatment of hemangioblastomas of the central nervous system in pediatric patients. AB - OBJECTIVE: Hemangioblastomas are histologically benign lesions that occur sporadically or as a manifestation of von Hippel-Lindau disease (VHL). The treatment strategy of these neoplasms is complicated by their unpredictable growth patterns and the often irreversible neurological deficits they may cause. This study aims to outline the neurosurgical treatment options and to address the ongoing debate of surgical timing in pediatric patients with VHL. PATIENTS AND METHODS: Thirteen consecutive pediatric patients (mean age 15.1 years) who were surgically treated for intracranial (n=8) or spinal hemangioblastomas (n=5) were included in this study (range of clinical follow up 12-86 months). Ten patients were affected by von Hippel-Lindau and three were with sporadic tumors. Serial examinations, preoperative MRI studies, and operative findings were reviewed. RESULTS: Patients with cerebellar and intramedullary hemangioblastomas did not develop additional neurological deficits postoperatively. Two patients with brainstem tumors exhibited transient hemiparesis and caudal nerve palsy, respectively. Both patients recovered completely from their postoperative deficits. Preoperatively symptomatic patients with spinal tumors did not deteriorate nor improve after surgery. During the observed follow-up periods, no tumor recurrences were observed. CONCLUSION: Central nervous system (CNS) hemangioblastomas in pediatric patients can be surgically treated with low morbidity. Based on our experience, we recommend considering also the surgical removal of asymptomatic hemangioblastomas with proven radiological progression to prevent the development of permanent neurological deficits. Molecular screening of every pediatric patient and family is mandatory to enable the detection of extraneurological tumors and the development of an efficient therapeutic strategy. PMID- 16369853 TI - Colloid cysts in children, a clinical and radiological study. AB - INTRODUCTION: Colloid cysts are uncommon in children. They occur more frequently in young adults. METHODOLOGY: This is a retrospective study on all pediatric patients ( or =60 years with low (T-score less than -1) lumbar bone mineral density (BMD). The hormone replacement therapy (HRT) group received a low dose of conjugated estrogen (CEE) at a dose of 0.31 mg/day +/- medroxyprogesterone acetate (MPA) 2.5 mg/day. Group HRT/D received the same dose of HRT together with alfacalcidol in a daily dose of 1.0 microg/day. Changes in lumbar BMD measured by dual energy X-ray absorptiometry (DXA) were followed every 6 months for 2 years. The lumbar BMD of group HRT increased 3.37% [95% confidence interval (CI) 1.6%-5.2%], 4.00% (95%CI 1.6%-6.4%), and 2.32% (95%CI -0.7% to 5.3%) at 12, 18, and 24 months, respectively, when the baseline value was taken as 0%. Lumbar BMD of group HRT/D showed a significant increase beyond 6 months. The percent increases for this group at 6, 12, 18, and 24 months were 6.18 (95%CI 1.3%-6.6%), 6.18% (95%CI 3.9%-8.5%), 7.17% (95%CI 4.3%-10.0%), and 8.75% (95%CI 6.0%-11.5%), respectively. In addition, there was a significant difference in the changes of the lumbar BMD between the two groups at 24 months, suggesting that the combination of HRT and alfacalcidol is more effective than HRT alone in terms of the BMD effect. This study is the first prospective trial demonstrating an additive effect of alfacalcidol on lumbar BMD in postmenopausal women receiving low-dose HRT. It suggests that the combination therapy can be considered to be a promising mode of treatment for bone loss after menopause. PMID- 16369893 TI - Effects of anti-parathyroid hormone-related protein monoclonal antibody and osteoprotegerin on PTHrP-producing tumor-induced cachexia in nude mice. AB - We have previously demonstrated that parathyroid hormone-related protein (PTHrP) is a cachexia inducer, but it is still not known what PTHrP effects on target tissues induce the cachexia. Therefore, we examined the effects of anti-PTHrP antibody and osteoprotegerin (OPG) on PTHrP-producing tumor-induced cachexia. Nude mice bearing PTHrP-producing human lung cancer cells (HARA-B) exhibited cachexia with hypercalcemia 3-4 weeks after inoculation, accompanied by losses in body, adipose tissue, and muscle weight. OPG ameliorated hypercalcemia, as did neutralization of PTHrP with antibody; and it increased both body and adipose tissue weights. These increases in body and adipose tissue weight, however, were significantly less than those in mice treated with anti-PTHrP antibody. Simultaneous administration of OPG and anti-PTHrP antibody caused significant increases in body, adipose tissue, and muscle weight, along with an immediate decrease in blood ionized calcium levels. The increase in body weight was similar to that observed in mice treated with anti-PTHrP antibody alone, and the decrease in the blood ionized calcium levels was significantly greater than that in mice treated with OPG or anti-PTHrP antibody alone. These results suggest that an effect of PTHrP on target tissues other than hypercalcemia is involved in the development of cachexia. Expression of cachexia-inducing proinflammatory cytokines (interleukin-6 and leukemia inhibitory factor) is stimulated by PTHrP. This might be a mechanism by which PTHrP produces tumor-induced cachexia. It is also suggested that OPG and anti-PTHrP antibody synergistically act to ameliorate hypercalcemia, although the mechanism responsible for this is unclear. PMID- 16369894 TI - Experimental model of tooth movement by orthodontic force in mice and its application to tumor necrosis factor receptor-deficient mice. AB - Orthodontic tooth movement is achieved by mechanical loading; however, the biological mechanism involved in this process is not clearly understood owing to the lack of a suitable experimental model. In the present study, we established an orthodontic tooth movement model in mice using a Ni-Ti closed coil spring that was inserted between the upper incisors and the upper first molar. Histological examination demonstrated that the orthodontic force moved the first upper molar mesially without necrosis of the periodontium during tooth movement. The number of TRAP-positive osteoclasts on the pressure side significantly increased in a time-dependent manner. Quantitative real time-based reverse transcription polymerase chain reaction analysis demonstrated increased levels of mRNA for cathepsin K. Immunohistochemical staining revealed the expression of tumor necrosis factor-alpha (TNFalpha) in periodontium on the pressure side of the first molar during orthodontic tooth movement. When this tooth movement system was applied to TNF type 1 receptor-deficient mice and TNF type 2 receptor deficient mice, tooth movement observed in TNF type 2 receptor-deficient mice was smaller than that in the wild-type mice and TNF type 1 receptor-deficient mice. The number of TRAP-positive osteoclasts on the pressure side was significantly small in TNF type 2 receptor-deficient mice compared with that in TNF type 1 receptor-deficient mice on day 6 after application of the appliance. The present study indicates that TNFalpha signaling plays some important roles in orthodontic tooth movement. PMID- 16369895 TI - CD38 is associated with premenopausal and postmenopausal bone mineral density and postmenopausal bone loss. AB - One goal of osteoporosis research is to identify the genes and environmental factors that contribute to low bone mineral density (BMD) and fracture. Linkage analyses have identified quantitative trait loci (QTLs), however, the genes contributing to low BMD are largely unknown. We examined the potential association of an intronic polymorphism in CD38 with BMD and postmenopausal bone loss. CD38 resides in 4p15, where a QTL for BMD has been described. CD38-/- mice display an osteoporotic phenotype at 3 months, with normalization of BMD by 5 months. The CD38 polymorphism was identified by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis in 457 postmenopausal and 173 premenopausal Caucasian women whose spine and hip BMD was measured by dual energy X-ray absorptiometry (DXA). Influence of the CD38 polymorphism on bone loss was analyzed in 273 postmenopausal women over a follow up of 2.94 +/- 1.50 years. The CD38-PvuII polymorphism was significantly associated with premenopausal and postmenopausal (P = 0.001) lumbar spine BMD. Women homozygous for the G allele had >14% lower spinal BMD than women with GC/CC genotypes. An allele dose effect was observed at the spine in premenopausal (P = 0.002) and postmenopausal (P < 0.001) cohorts. The CD38-PvuII polymorphism was significantly associated with femoral neck BMD in pre- and postmenopausal women (P = 0.002 and P = 0.011, respectively). However, significance was lost following adjustment of hip BMD for covariates in the postmenopausal cohort (P = 0.081). The CD38-PvuII polymorphism was weakly associated with bone loss at the spine (P = 0.024), in postmenopausal women not taking hormone replacement therapy. We suggest that the CD38-PvuII polymorphism may influence the attainment and maintenance of peak BMD and postmenopausal bone loss. PMID- 16369896 TI - Tests of linkage and association of PTH/PTHrP receptor type 1 gene with bone mineral density and height in Caucasians. AB - Parathyroid hormone/parathyroid hormone-related peptide receptor type 1 (PTHR1) plays an important role in calcium metabolism. It was previously shown to influence variation in bone mineral density (BMD). To investigate its importance in a typical U.S. Caucasian population, we tested linkage or association of the PTHR1 gene with BMD and height. Altogether, 1873 subjects from 405 Caucasian nuclear families were studied. BMD was measured at the lumbar spine (L1-L4) and total hip (femoral neck, trochanter, and intertrochanter regions). Four single nucleotide polymorphisms (SNPs) in the PTHR1 gene were genotyped. Sixteen haplotypes were reconstructed. Only two major haplotypes had frequencies >3% and were thus used for the analysis. Analyses were performed for BMD and height in the total sample and for peak BMD (PBMD) achieved in offspring subjects aged 20 50 in a subsample of 387 nuclear families. We found suggestive evidence for total association between haplotype 13 (AATG) and hip PBMD (P = 0.031). For height, evidence of within-family association was suggested for SNP1, SNP2, and haplotype 4 (GGCA) (P < or = 0.05). Our findings suggest that the PTHR1 gene may be important for PBMD, height variation, or both, although the significance is dampened by correction for multiple testing. PMID- 16369897 TI - 25-hydroxyvitamin D, parathyroid hormone, and functional recovery after hip fracture in elderly patients. AB - There is increasing interest in the effects of vitamin D and parathyroid hormone (PTH) on extraskeletal tissues, including the muscle. These effects may explain impairment in functional ability found in vitamin D-deficient subjects. Our aim was to investigate the roles of vitamin D and PTH in affecting the ability to perform activities of daily living after hip fracture. We studied 456 of 521 hip fracture patients admitted consecutively to a rehabilitation hospital. Functional outcome was assessed after acute inpatient rehabilitation by using the Barthel index score. The functional scores were significantly correlated with serum levels of 25-hydroxyvitamin D (rho = 0.190; P < 0.001) and PTH (rho = -0.164; P < 0.001) and the 25-hydroxyvitamin D/PTH ratio (rho = 0.261; P < 0.001). At multiple regression, 25-hydroxyvitamin D and PTH levels were independently associated with Barthel index scores. The correlation between the 25 hydroxyvitamin D/PTH ratio and Barthel index scores was significantly stronger than the one between 25-hydroxyvitamin D and Barthel index scores (difference between the two correlation coefficients = 0.071; 95% CI = 0.009-0.133; P = 0.011). The significant association between the 25-hydroxyvitamin D/PTH ratio and the Barthel index scores persisted after adjustment for 12 prognostic factors (P = 0.012). On the whole, the panel of prognostic factors we studied predicted 50.1% of the variance of the functional score. Data shows that PTH and 25 hydroxyvitamin D were significantly associated with the ability to function after hip fracture and suggest that the two hormones act through independent mechanisms. The 25-hydroxyvitamin D/PTH ratio significantly contributed to a predictive model of functional outcome. PMID- 16369898 TI - Genetic and enzymatic analysis for two Japanese patients with idiopathic infantile arterial calcification. AB - Idiopathic infantile arterial calcification (IIAC) is a life-threatening disorder in young infants. Cardiovascular symptoms are usually apparent within the first month of life. The symptoms are caused by calcification of large and medium-sized arteries, including the aorta, coronary arteries, and renal arteries. Most of the patients die by 6 months of age because of heart failure. Recently, homozygous or compound heterozygous mutations for the ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) gene were reported as causative for the disorder. ENPP1 regulates extracellular inorganic pyrophosphate (PPi), a major inhibiter of extracellular matrix calcification. Two Japanese patients with IIAC were studied. One, from first-cousin parents, showed a typical clinical course. The onset in the second patient was late. Both of the patients were clinically compatible for IIAC; arterial calcification was shown, and hypertension was prominent. We sequenced all the exons and exon-intron boundaries of the gene and measured nucleotide pyrophosphohydrolase (NPPH) activity of ENPP1. Homozygous Arg730Stop was detected in the typical IIAC patient. The mutation was a novel nonsense mutation and not detected in 60 healthy controls. His NPPH activity was 4% of normal. On the other hand, the late-onset patient was not shown to have any mutations. NPPH activity in this patient was 70% of normal. We confirmed that ENPP1 was also responsible for the Japanese patient with IIAC. The atypical late-onset phenotype may not be associated with ENPP1 abnormalities. IIAC is considered to be a clinically and genetically heterogeneous disorder. PMID- 16369899 TI - Relations between interleukin-1, its receptor antagonist gene polymorphism, and bone mineral density in postmenopausal Korean women. AB - We investigated the relation between polymorphisms in the interleukin-1(IL-1) and IL-1 receptor antagonist (IL-1ra) gene, and bone mineral density (BMD) in postmenopausal Korean women. The IL-1alpha C(-889)T polymorphism, and IL-1beta C( 511)T polymorphism were examined by restriction fragment length polymorphism, and 86-bp variable number tandem repeat polymorphism in the IL-1ra gene was analyzed by the polymerase chain reaction and electrophoresis in 202 postmenopausal Korean women. Serum osteocalcin, and C-telopeptide of type I collagen were measured using a radioimmunoassay and enzyme-linked immunosorbent assay, respectively. BMD at the lumbar spine and proximal femur was measured by dual-energy X-ray absorptiometer. No significant differences in BMD or in serum bone markers levels were noted across the IL-1alpha or IL-1beta genotype. There were no significant differences in the distribution of IL-1alpha or IL-1beta genotype according to the status of bone mass. BMD in women carrying the A2 allele of the IL-1ra gene was significantly lower than those without this allele, and the A2 allele was more frequent in osteoporotic women than in normal women. These data suggest that IL-1ra gene VNTR polymorphism is a genetic factor that may affect BMD in Korean women. PMID- 16369900 TI - Causes of secondary hyperparathyroidism in a healthy population: the Tromso study. AB - Secondary hyperparathyroidism (SHPT) develops as a compensatory mechanism when the body is in calcium deficit. SHPT may be harmful and has been associated with elevated blood pressure. The cause of SHPT could be low calcium intake, reduced intestinal calcium absorption, or increased excretion. However, the relative importance of these factors for the development of SHPT is not known. During the 5th Tromso study, serum PTH and calcium were measured in 7954 subjects. Then 96 subjects with SHPT (defined as serum PTH above 6.4 pmol/l together with serum calcium below 2.40 mmol/l) and 106 control subjects were examined at follow-up with a food frequency questionnaire, calcium absorption test, measurement of 24-h urinary calcium excretion, and serum vitamin D status. The statistical analyses showed several interactions necessitating subgroup analysis. It was found that the calcium intake was significantly lower in the SHPT group, but only in nonsmoking males; the calcium absorption was nonsignificantly higher in the SHPT group; the serum 25-hydroxyvitamin D levels were significantly lower in the SHPT group but only in nonsmokers; and the 24-h urinary calcium excretion was significantly lower in the SHPT group but only in those not on blood pressure medication. The most frequent cause of SHPT appeared to be low calcium intake (18%) and a low serum 25-hydroxyvitamin D level (18%). However, in most subjects with SHPT all tests were within the normal range, and the cause is therefore probably a combination of several factors. PMID- 16369901 TI - Endogenous sex steroids and bone mineral density in healthy Greek postmenopausal women. AB - The aim of this study was to assess the association of endogenous sex steroids with bone mineral density (BMD) in healthy postmenopausal women not on hormone therapy. A total of 884 postmenopausal women aged 42-71 years were studied in a cross-sectional design. Parameters assessed were follicle-stimulating hormone, luteinizing hormone, estradiol, total testosterone, sex hormone-binding globulin, free estrogen index (FEI), free androgen index (FAI), Delta4-androstendione (Delta4A), dehydroepiandrosterone sulfate (DHEAS), bone alkaline posphatase, and bone mineral density at the lumbar spine (L-BMD) and femoral neck (N-BMD). Estradiol and FEI associated positively with both L-BMD and N-BMD (r = 0.21-0.47, P < 0.01). These associations remained significant after adjustment for age, years since menopause, and body mass index. FAI correlated positively with both L BMD and N-BMD (r = 0.18 and 0.33, respectively; P < 0.01). At the multivariate analysis, however, FAI remained the significant determinant only for N-BMD. Delta4A associated positively with N-BMD (r = 0.27, P = 0.001), whereas DHEAS showed no association with BMD at either site. Thus, endogenous steroids are significant determinants of postmenopausal BMD. Endogenous estradiol may be more important for lumbar spine BMD, whereas endogenous androgens are associated mainly with femoral neck BMD. PMID- 16369902 TI - Bone mineral density and turnover in patients with acromegaly in relation to sex, disease activity, and gonadal function. AB - Acromegaly is a rare disease caused by growth hormone (GH) hypersecretion. GH and insulin-like growth factor-I (IGF-I) exert anabolic activity in bones. Nevertheless, bone mineral density (BMD) loss is not uncommon in patients with acromegaly. It is assumed to be due to hypogonadism associated with the acromegaly. The aim of the study was to examine BMD at various skeletal sites and bone turnover and to assess the influence of impaired gonadal function and disease activity on BMD and turnover changes in acromegaly. A total of 62 patients were studied (40 women, 22 men). Among the women, 22 had active disease and 18 were cured; 16 women had normal gonadal function, and 24 were hypogonadal. Altogether, 12 men presented with active acromegaly, and 10 were cured; normal gonadal function was found in 10 men, and hypogonadism was diagnosed in 12 men. Controls were 30 healthy subjects. Densitometry using dual-energy X-ray absorptiometry of the lumbar spine, proximal femur, forearm, and total body was carried out. Bone turnover was studied based on serum osteocalcin, C-terminal collagen type 1 crosslinks, and bone alkaline phosphatase concentration. A disadvantageous effect of acromegaly on bone density was associated with hypogonadism in the distal radius (in women), the proximal femur (in men), and the total body (both sexes). An anabolic effect of GH during active acromegaly was present in the proximal femur only in men. We confirmed increased bone turnover in the presence of acromegaly, and these changes were similar regarding the activity of the disease and the gonadal status. PMID- 16369903 TI - Bone mineral density and its correlation with clinical and laboratory factors in chronic peritoneal dialysis patients. AB - The aim of this study was to assess the clinical and laboratory correlations of bone mineral density (BMD) measurements among a large population of patients on chronic peritoneal dialysis (PD). This cross-sectional, multicenter study was carried out in 292 PD patients with a mean age of 56 +/- 16 years and mean duration of PD 3.1 +/- 2.1 years. Altogether, 129 female and 163 male patients from 24 centers in Canada, Greece, and Turkey were included in the study. BMD findings, obtained by dual-energy X-ray absorptiometry (DEXA) and some other major clinical and laboratory indices of bone mineral deposition as well as uremic osteodystrophy were investigated. In the 292 patients included in the study, the mean lumbar spine T-score was -1.04 +/- 1.68, the lumbar spine Z-score was -0.31 +/- 1.68, the femoral neck T-score was -1.38 +/- 1.39, and the femoral neck Z score was -0.66 +/- 1.23. According to the WHO criteria based on lumbar spine T-scores, 19.2% of 292 patients were osteoporotic, 36.3% had osteopenia, and 44.4% had lumbar spine T-scores within the normal range. In the femoral neck area, the prevalence of osteoporosis was slightly higher (26%). The prevalence of osteoporosis was 23.3% in female patients and 16.6% in male patients with no statistically significant difference between the sexes. Agreements of lumbar spine and femoral neck T-scores for the diagnosis of osteoporosis were 66.7% and 27.3% and 83.3% for osteopenia and normal BMD values, respectively. Among the clinical and laboratory parameters we investigated in this study, the body mass index (BMI) (P < 0.001), daily urine output, and urea clearance time x dialysis time/volume (Kt/V) (P < 0.05) were statistically significantly positive and Ca x PO(4) had a negative correlation (P < 0.05) with the lumbar spine T scores. Femoral neck T scores were also positively correlated with BMI, daily urine output, and KT/V; and they were negatively correlated with age. Intact parathyroid hormone levels did not correlate with any of the BMD parameters. Femoral neck Z scores were correlated with BMI (P < 0.001), and ionized calcium (P < 0.05) positively and negatively with age, total alkaline phosphatase (P < 0.05), and Ca x P (P < 0.01). The overall prevalence of fractures since the initiation of PD was 10%. Our results indicated that, considering their DEXA based BMD values, 55% of chronic PD patients have subnormal bone mass-19% within the osteoporotic range and 36% within the osteopenic range. Our findings also indicate that low body weight is the most important risk factor for osteoporosis in chronic PD patients. An insufficient dialysis dose (expressed as KT/V) and older age may also be important risk factors for osteoporosis of PD patients. PMID- 16369904 TI - Rapid pain relief and remission of sternocostoclavicular hyperostosis after intravenous ibandronate therapy. AB - Sternocostoclavicular hyperostosis (SCCH) is an infrequent but painful, localized disturbance of bone metabolism of unknown etiology. The diagnosis of SCCH is generally one of exclusion, and it is therefore frequently missed or delayed, leaving patients with pain that frequently fails to respond to standard analgesic therapy. Consequently, SCCH leads to significantly impaired quality of life. Characteristic increased localized bone turnover and inflammatory osteitis provide a strong rationale for using intravenous bisphosphonates to treat the condition. We report on three patients with long-standing, treatment-refractory SCCH in whom intravenous ibandronate injections (a single administration of 4 mg followed by 2 mg every 3 months for up to a year) produced prompt, dramatic, persistent pain relief and resolution of the other symptoms of the disease. We also review recent evidence suggesting that SCCH is more common than generally believed and that technetium-99 bone scanning can aid in making an accurate diagnosis. PMID- 16369905 TI - [Ultrasound evaluation of the acromioclavicular joint. A comparison with magnetic resonance imaging]. AB - INTRODUCTION: Ultrasound is useful in detecting acromioclavicular pathologies in cases of trauma, inflammations and degenerative changes in sports medicine. Many studies compare joint space and capsular dimensions of symptomatic and asymptomatic patients. However, no study has examined the reproducibility and reliability of these measurements. The aim of this study was to evaluate the reliability of ultrasonographic measurements in assessing the acromioclavicular joint. MATERIALS AND METHODS: 27 acromioclavicular joints of 15 healthy subjects were examined by T1 weighted magnetic resonance imaging (MRI) to determine the normal limits of joint space (a) and joint capsule (b). These measurements were compared to standardised ultrasonographic (11 Mhz) measurements, which were repeated three times. RESULTS: The mean difference between MRI and ultrasound measurements was 1.5 +/- 1.3 mm and 1.3 +/- 1.2 mm for distance a and b, respectively. Reproducibility of ultrasonographic measurements was high with a mean standard deviation of 0.3 +/- 0.2 mm and 0.4 +/- 0.3 mm for distance a and b, respectively. CONCLUSION: Due to low costs, safety and wide availability ultrasonography is suited for the evaluation of the acromiocalvicular joint. However, when interpreting the results measurement errors, limitations in resolution of the system used, and the anatomy of the acromioclavicular joint and its anatomic variants have to be taken into consideration. PMID- 16369906 TI - [Delayed repair of traumatic shoulder girdle muscle tears]. AB - Muscle ruptures effecting the shoulder girdle represent a comparatively rare event. Especially after delayed diagnostics, the operative result is only heavily predictable. Due to few case reports, there are no experiences to guide us. This case report describes the delayed surgical treatment of two cases of traumatic shoulder girdle muscle tears. These results encourage, that also after a long time period operative treatment can lead to a good result. PMID- 16369907 TI - [Stress-induced changes of attention during choking in judo]. AB - The aim of the investigation is the registration and analysis of cognitive parameters in the alarm phase of the stress reaction during a conventional judo cross choking technique. 57 judoka (41 males, 16 females; mean age 23.1 +/- 6.7 years) underwent a standardized cross-choking maneuvre (juji-jime), which lasted for 6 - 8 s until giving up, carried out by a black belt physician. The Stroop inference test was performed before and twice after the intervention. The reading velocity of the interference task was reduced by an average of 77.4 s to a mean of 74.4 s (p < 0.01), and the number of reading errors decreased by a mean from 2.9 to 2.1 (p < 0.01), finally increasing to 2.4 s. The data show the most important improvement in performance in 25 s after choking in the male judoka. In the female judoka increasing performance started already 25 s before choking and continued 25 s after the intervention. The alarm reaction following the potential life threatening stressor with an attack upon the throat improves selective attention before (female subjects) and after choking (female and male subjects) depending on gender. PMID- 16369908 TI - [Risk of injury in and adolescent and junior competitive karate]. AB - Increasing professionalism in competitive karate has led to a specific injury pattern. The aim of our study was to investigate the injury pattern in junior competitive karate and to evaluate the prophylactic effect of fist padding. During the 4 (th) Junior World Championships of the World Karate Confederation (WKC) 2004 235 karate fights were observed and injury-related data were recorded. Fist padding was used in 183 fights, 52 fights were carried out without. In 22 % of all fights we saw an injury. Most of them were mild injuries of the head and face: 32 bruises of the face (13 with epistaxis), 7 facial lacerations and 3 concussions (mild brain injury). 14 fights were stopped due to an injury. Only in the category male 18 - 20 years a higher rate of injuries was seen without fist padding (shobu ippon) compared to shobu sanbon (with fist padding). The injury pattern of junior competitive karate is comparable to senior karate. Fist padding does not generally reduce the incidence or severity of injuries. PMID- 16369909 TI - [The trend sport snowblading and its risks]. AB - The study analysed the results of an examination of 3557 skiing, snowboarding and snowblading injuries treated at the Department of Trauma Surgery at the Klinikum Garmisch-Partenkirchen in winter seasons 99/00 through 02/03. In this study group, a total of 70 injuries occurred while snowblading. These injuries are compared to the remaining injuries resulting from skiing, carving or snowboarding accidents. This study intends to contribute to the tracking and recording of injuries occurring during snowblading, a sport that is popular but not yet well addressed in medical literature. The results show that snowblading injuries are similar to those of skiing, but different from those attributable to snowboarding, particularly with regard to the affected parts of the body and injury patterns. Acrobatic jumps increase significantly the risk of fractures of the lower extremities. However, ruptures of ligaments, especially of the ACL, are seen more rarely in snowblading than in carving or conventional skiing. Distorsions, on the other hand, are observed significantly more often among snowbladers when compared with carvers and snowboarders. Release bindings for snowblades could reduce the risk of injuries. PMID- 16369910 TI - [Treatment of incomplete meniscal lesions in athletes]. AB - PURPOSE: Arthroscopic treatment of complete meniscal lesions is well established. Nevertheless there is discussion, how to treat incomplete meniscal tears, especially in younger and active patients. This study was designed to evaluate our standard-therapy without refixation of the meniscus. METHOD: Between 7/89 and 3/01 in 47 patients (48 knees, O age 29 years) an incomplete meniscal lesion following sports injury was found. The lesions were revitalized by "needling" or shaving. We performed no refixation. All patients had a postoperative flexion limit in an orthosis for 6 weeks. The follow-up examination was performed 6.5 (2 14) years postoperative. RESULTS: The avarage Lysholm-Score increased significantly from 55 points preoperative to 92 points at the follow-up examination. The Tegner-Score increased from 3.3 points preoperative to 6.2 points at the follow-up examination. The overall result was rated "exellent" and "good" by 83 % of the patients, "fair" by 15 % of the patients and "poor" by 2 % of the patients. Return to sports activity was possible at an avarage of 7 (3 - 12) months postoperative. CONCLUSIONS: In summary we found, that shaving and "needling" of an incomplete meniscal lesion in combination with partial synovectomi and standardized postoperative treatment leads to a high healing rate. A limited flexion for 6 weeks postoperativ in an orthosis at full weight bearing is recommended. In case of complete healing of the lesion the chondroprotective and joint stabilizing function of the meniscus, especially in young and active patients is obtained. PMID- 16369911 TI - [Diagnosis of stress fractures in adolescents]. AB - Stress fractures in adults are a well described and frequently observed overuse injury with good clinical outcome. In adolescents, stress fractures are very rare and there are only few studies devoted exclusively this type of injury in patients with immature skeletal system. Prognosis of stress fractures in adolescents mainly depends on early diagnosis and early initiated therapy. The object of the present study is to describe typical clinical symptoms as well as characteristic radiological findings in adolescent patients with stress fractures. Thirty-five patients with a total of 40 stress fractures affecting bone with the growth plates still open have been included in the present study. The average duration of follow-up was 4.7 years (standard deviation +/- 2.45, with a minimum follow of 12 months). The average age at occurrence was 12 years (SD +/- 4.31). At initial presentation, pain at exposure was declared as cardinal symptom in 19 of 40 cases, while pain at rest has been described in only 9 cases. An additional swelling has been noticed in 12 cases. Nevertheless, all patients described pain as their major complain. Conventional radiography was unconspicious in 12 of 40 cases; periost hypertrophy was found in 14 cases, while in further 14 cases a fracture line could be detected in conventional radiography. In all cases, in which a magnetic resonance imaging (MRI) has been conducted, changes in signal intensity were detected (including all cases of unconspicous x-ray). Our data indicate, that especially in young sportsmen with pain at exposure of the lower extremity, even if conventional radiography does not reveal abnormalities, MRI should be conducted to detect stress fractures of the juvenile skeletal system in early stages. Only if diagnosis is made early, consistent therapy can be applied as early as needed in order to achieve good clinical results. PMID- 16369912 TI - [Traumatic internal carotid artery dissection associated with triathlon: a rare differential diagnosis]. AB - Most cervical artery dissections occur spontaneously, but traumatic mechanisms can occasionally be identified. Traumatic internal carotid artery dissection has been attributed with several sports, but there are no reports of its occurrence in thriathlon. We report the case of a 63-year-old semi-professional triathlete, who noticed right-sided neck pain and parietooccipital headache after a triathlon competition. Neurological examination revealed an incomplete right Horner's syndrome. MRI and MR-angiography demonstrated a right-sided internal carotid artery dissection with segmental loss of flow-signal and a mural haematoma in the vessel wall. In this case two pathophysiological mechanisms may be considered. Our patient reported receiving several blows to the neck in a congested 50-metre pool during the triathlon swimming stage. Furthermore, the hyperextended position of the neck during the subsequent cycling stage may have contributed, to a large extent, to the arterial dissection. The risk factors, presenting symptoms, diagnosis of internal carotid artery dissection are reviewed. PMID- 16369913 TI - The effect of CP96,345 on the expression of tachykinins and neurokinin receptors in acute pancreatitis. AB - Acute pancreatitis (AP) is a life-threatening condition that involves an acute inflammatory process in the pancreas. The involvement of tachykinins and neurokinin receptors in acute pancreatitis has been described only recently, despite their long-established role in inflammatory conditions. Among these, substance P (SP) is believed to play a central role in exacerbating the inflammatory process by acting through neurokinin-1 receptor (NK1R). Treatment with the NK1R antagonist, CP96,345, results in protection against caerulein induced acute pancreatitis in mice. However, the mechanism by which NK1R and SP worsen the condition is still unclear. In the present study, we have investigated the effect of NK1R blockage on the expression of preprotachykinin genes and neurokinin receptors in acute pancreatitis. In the pancreas, CP96,345 treatment resulted in suppression of the elevation of SP concentration, preprotachykinin-A gene (PPT-A) mRNA expression, and NK1R mRNA and protein expression. In the lungs, the antagonist was found to suppress the increase in SP concentration, PPT-A mRNA expression and preprotachykinin-C gene (PPT-C) mRNA expression. However, the antagonist treatment further promoted the accumulation of pulmonary NK1R mRNA and protein expression. Neurokinin-2 receptor (NK2R) mRNA expression was not detected in normal pancreas. However, up-regulated expression of the mRNA for this receptor was observed during acute pancreatitis and treatment with CP96,345 further increased this expression. Pulmonary NK2R mRNA expression was found to be reduced during acute pancreatitis and CP96,345 treatment normalized this reduction. Neurokinin-3 receptor (NK3R) mRNA expression was absent in both pancreas and lung. These data have provided valuable information regarding the regulation of tachykinins and neurokinin receptors during acute pancreatitis. PMID- 16369914 TI - The p38 transduction pathway in prostatic neoplasia. AB - It has been proposed that, among other cellular responses, TNF-alpha induces not only cell death, but also cell proliferation by activation of p38. It has also been reported that IL-1-alpha favours cell proliferation by p38 activation. The aim of the present study was to evaluate upstream (alpha-PAK, MEK-6) and downstream (Elk-1 and ATF-2) components of the p38 transduction pathway in normal prostate, benign prostatic hyperplasia (BPH), and prostate carcinoma (PC). Immunohistochemical and western blot analyses were performed in 20 samples of normal prostate, 47 samples of BPH, and 27 samples of PC. In all normal prostates, immunoreactivity for p-Elk-1 and p-ATF-2 was observed in epithelial cell nuclei, but no expression of alpha-PAK or MEK-6. In BPH, there was expression of alpha-PAK (cytoplasm) and MEK-6 (cytoplasm), while the proportions of lesions that were immunoreactive for p-Elk-1 (nucleus and cytoplasm) and p-ATF 2 (nucleus) decreased. In PC, the percentages of cells that were immunoreactive for alpha-PAK (cytoplasm) or MEK-6 (cytoplasm) rose slightly in comparison with BPH, while the percentages of cells that were immunoreactive for p-Elk-1 (nucleus and cytoplasm) or p-ATF-2 (nucleus and cytoplasm) were much higher than in BPH. It is concluded that overexpression of alpha-PAK, MEK-6, p38, p-Elk-1, and p-ATF 2 in BPH, and more intensely in PC, enhances cell proliferation. In BPH, such proliferation is triggered by IL-1 and in part counteracted by the TNF-alpha/AP-1 pathway, which promotes apoptosis. In PC, proliferation is triggered by IL-1 and TNF-alpha (the TNF-alpha/AP-1 pathway is diverted towards p38 activation). Since in a study of the same patients immunoexpression of IL-1alpha and IL-1RI was previously observed to be increased in PC, inhibition of p38 is a possible target for PC treatment, as this inhibition would both decrease IL-1-induced cell proliferation and increase TNF-alpha-induced cell death. PMID- 16369915 TI - RNA interference reveals a role for TLR2 and TLR3 in the recognition of Leishmania donovani promastigotes by interferon-gamma-primed macrophages. AB - Leishmania donovani promastigotes evade the induction of a proinflammatory response during their invasion of naive macrophages. However, their entry into IFN-gamma-primed macrophages is accompanied by the secretion of nitric oxide (NO) and proinflammatory cytokines. In the present study, we addressed the hypothesis that priming with IFN-gamma induces the expression of a receptor that enables mouse macrophages to recognize L. donovani promastigotes. We observed that in IFN gamma-primed macrophages, L. donovani promastigotes stimulated Interleukin-1 receptor-associated kinase-1 (IRAK-1) activity. We next showed that Toll-like receptor (TLR)3 is barely detectable in naive macrophages but is expressed in IFN gamma-treated macrophages. Silencing of TLR3, TLR2, IRAK-1 and myeloid differentiation factor 88 (MyD88) expression by RNA interference revealed that both TLR are involved in the secretion of NO and TNF-alpha induced by L. donovani promastigotes. Using L. donovani mutants, we showed that TLR2-mediated responses are dependent on Galbeta1,4Manalpha-PO(4)-containing phosphoglycans, whereas TLR3 mediated responses are independent of these glycoconjugates. Furthermore, our data indicate a participation of TLR2 and TLR3 in the phagocytosis of L. donovani promastigotes and a role for TLR3 in the leishmanicidal activity of the IFN-gamma primed macrophages. Collectively, our data are consistent with a model where recognition of L. donovani promastigotes depends on the macrophage activation status and requires the expression of TLR3. PMID- 16369916 TI - Time-dependent resveratrol-mediated mRNA and protein expression associated with cell cycle in WR-21 cells containing mutated human c-Ha-Ras. AB - Cancer results from an undesirable imbalance between cellular proliferation and apoptosis. Both processes may be modulated at the level of gene expression, viz., p53 and c-Ha-ras, by dietary bioactive components such as resveratrol. We tested the time-dependent effect of resveratrol on gene and protein expression in WR-21 cells containing a mutated human c-Ha-ras oncogene. We demonstrate cyclic resveratrol-mediated expression of p53, mdm2, p21(cip/waf), Rb, and cyclin G at both the RNA and the protein level at <8 h. However, ras was not differentially expressed at either the RNA or the protein level. p53 was upregulated followed by p21cip/waf, then mdm2, and cyclin G, all downstream p53-activated targets. RNA transcription increased at >8 h for all genes except p53, but protein levels did not suggest uncoupling of transcription and translation. At 24 h, both p53 and Rb expression returned to baseline, suggesting collapse of DNA structure and spindle assembly checkpoints characteristic of mitotic catastrophe. In summary, resveratrol at <8 h induced p53-mediated effects, including apoptosis and cell cycle arrest (G2/M). However, later, it induced cell-cycle checkpoint dysfunction, indicative of mitotic catastrophe. Thus, future studies should better elucidate the temporal mechanism of the dietary bioactive agent resveratrol on cancer cells. PMID- 16369917 TI - Advances in locally constrained k-space-based parallel MRI. AB - In this article, several theoretical and methodological developments regarding k space-based, locally constrained parallel MRI (pMRI) reconstruction are presented. A connection between Parallel MRI with Adaptive Radius in k-Space (PARS) and GRAPPA methods is demonstrated. The analysis provides a basis for unified treatment of both methods. Additionally, a weighted PARS reconstruction is proposed, which may absorb different weighting strategies for improved image reconstruction. Next, a fast and efficient method for pMRI reconstruction of data sampled on non-Cartesian trajectories is described. In the new technique, the computational burden associated with the numerous matrix inversions in the original PARS method is drastically reduced by limiting direct calculation of reconstruction coefficients to only a few reference points. The rest of the coefficients are found by interpolating between the reference sets, which is possible due to the similar configuration of points participating in reconstruction for highly symmetric trajectories, such as radial and spirals. As a result, the time requirements are drastically reduced, which makes it practical to use pMRI with non-Cartesian trajectories in many applications. The new technique was demonstrated with simulated and actual data sampled on radial trajectories. PMID- 16369918 TI - Lung function tests in neonates and infants with chronic lung disease: global and regional ventilation inhomogeneity. AB - This review considers measurement of global and regional ventilation inhomogeneity (VI) in infants and young children with acute neonatal respiratory disorders and chronic lung disease of infancy (CLDI). We focus primarily on multiple-breath inert gas washout (MBW) and electrical impedance tomography (EIT). The literature is critically reviewed and the relevant methods, equipment, and studies are summarized, including the limitations and strengths of individual techniques, together with the availability and appropriateness of any reference data. There has been a recent resurgence of interest in using MBW to monitor lung function within individuals and between different groups. In the mechanically ventilated, sedated, and paralyzed patient, VI indices can identify serial changes occurring following exogenous surfactant. Similarly, global VI indices appear to be increased in infants with CLDI and to differentiate between infants without lung disease and those with mild, moderate, and severe lung disease following preterm birth. While EIT is a relatively new technique, recent studies suggest that it is feasible in newborn infants, and can quantitatively identify changes in regional lung ventilation following alterations to ventilator settings, positive end expiratory pressure (PEEP), and administration of treatments such as surfactant. As such, EIT represents one of the more exciting prospects for continuous bedside pulmonary monitoring. For both techniques, there is an urgent need to establish guidelines regarding data collection, analysis, and interpretation in infants both with and without CLDI. PMID- 16369919 TI - Management of hyperplastic nephroblastomatosis. PMID- 16369920 TI - Models of care for survivors of childhood cancer. AB - With improvements in therapy for childhood cancer, the expectation that most childhood cancer patients will survive and enter adulthood is a reality. There is clear evidence that survivors are at risk for adverse health-related long-term sequelae associated with their cancer and its treatment, requiring appropriate health care resources. What is less clear is how this health care should optimally be delivered. We review the functional and operational needs for long term follow-up for childhood cancer survivors and present alternatives for models of care. Programs for childhood cancer survivors should provide mechanisms for monitoring and management of late effects, as well as support and advocacy for addressing psychosocial issues, health education, and assistance with financial concerns. Access to research is an important component as clinical care and research are integrally related. A multidisciplinary model that provides continuity of care throughout the disease course is optimal, providing transitions from acute anti-neoplastic therapy to follow-up and primary care, as well as from pediatric care to adult-oriented care. There is no single best model of care for all childhood cancer survivors. In evaluating different models, considerations include available resources as well as the particular cancer population being served. Not all survivors require the same level of services and the service level requirement for individual patients may change with time. As outcome research progresses for childhood cancer survivors, methodological issues of optimal health care delivery for this population deserve to be the subject of such research. PMID- 16369921 TI - Barriers to follow-up care of survivors in the United States and the United Kingdom. AB - Survivors of pediatric cancer face an increased risk of morbidity, mortality, and diminished quality of life associated with their previous cancer therapy. Because these risks are often modifiable, periodic lifetime medical follow-up is recommended for all survivors. This article assesses the current status of follow up care in the United States and the United Kingdom. Potential barriers in both cultures and health care environments are described. A better understanding of these barriers has led to the development and testing of interventions intended to optimize the health care of survivors. PMID- 16369923 TI - Growth hormone deficiency after chemotherapy for acute lymphoblastic leukemia in children who have not received cranial radiation. AB - Chemotherapy-related growth failure is a significant problem in children with acute lymphoblastic leukemia (ALL) and other childhood cancers. Growth impairment after cranial radiation (CR) can result in diminished adult height, but growth failure following chemotherapy without CR is usually followed by catch-up growth and normal adult height.1 A retrospective review of 347 ALL survivors registered in our Long Term Follow Up (LTFU) Clinic, since 1997 revealed that 109 had received CR; 3, total body irradiation (TBI); and 235, neither CR nor TBI. For patients whose growth velocity slowed, growth hormone (GH) levels and pediatric endocrinology referrals were obtained. Among the 112 ALL survivors who had received some form of CR, 5 had significant growth failure with growth hormone deficiency (GHD). Among the 235 ALL survivors treated with chemotherapy without CR, 2 were diagnosed with growth failure and GHD. We report the two survivors of childhood ALL treated with chemotherapy without CR who required GH replacement due to absence of catch-up growth. A 15-year-old boy and a 12-year-old girl, off therapy for 9 and 6 years, respectively, were evaluated for decreased growth velocity and failure of catch-up growth. Peak GH responses to stimulation using arginine and clonidine were 3.4 and 3.0 ng/ml, respectively (normal >10 ng/ml). Other causes of growth failure were ruled out, and GH replacement therapy was instituted. Their chemotherapy had included methotrexate, 6 mercaptopurine, vincristine, adriamycin, cyclophosphamide, L-asparaginase, dexamethasone, cytarabine, 6 thioguanine, and intrathecal methotrexate. The growth of all children treated with intensive chemotherapy, regardless of whether CR was administered, should be closely monitored with measurement of standing height at 6 months intervals until growth is complete. PMID- 16369922 TI - Two infants with life-threatening diffuse neonatal hemangiomatosis treated with cyclophosphamide. AB - Diffuse neonatal hemangiomatosis (DNH) is a rare, frequently fatal disorder characterized by multiple cutaneous and visceral hemangiomas. Complications include high-output cardiac failure, hemorrhage, hepatic failure, and consumption coagulopathy. We present two cases of neonates with DNH. We prescribed cyclophosphamide after failure of corticosteroids. Cyclophosphamide induced a fast regression of the lesions with no side effects in both infants. We were able to stop treatment after 3-4 courses of cyclophosphamide in both infants because of the excellent response. Cyclophosphamide seems to be a safe and effective treatment in patients suffering from life-threatening DNH unresponsive to corticosteroids. PMID- 16369924 TI - Long-term follow-up of pediatric cancer survivors: education, surveillance, and screening. AB - Cancer and its treatment predispose childhood cancer survivors to chronic or late occurring health problems that may not become clinically significant until many years after therapy. Frequently, long-term survivors of childhood cancer report late cancer-related effects that diminish quality of life and increase the risk of early mortality. Risk-based health care that involves a personalized plan for surveillance, screening, and prevention is recommended to reduce cancer-related morbidity in childhood cancer survivors. To implement optimal risk-based care, the survivor and health care provider must have accurate information about cancer diagnosis, treatment modalities, and potential cancer-related health risks to guide screening and risk-reducing interventions. However, previous studies evaluating health knowledge of childhood cancer survivors demonstrate noteworthy deficits and misperceptions about their cancer diagnosis, treatment, and cancer related health risks. In addition, because of the relative rarity of childhood cancer, many health care providers lack familiarity with cancer-related health risks and risk-reduction methods relevant for this population. To correct these deficits, the Scottish Intercollegiate Guidelines Network (SIGN) and the Children's Oncology Group (COG) developed clinical practice guidelines to foster appropriate risk-based survivor care. Herein, we discuss the development, benefits, and limitations of the SIGN and COG guidelines and the foundation they provide for standardizing long-term follow-up care of the ever-growing vulnerable population of childhood cancer survivors. PMID- 16369925 TI - Acidic pH hyperpolarizes nasal potential difference. AB - Nasal potential difference (NPD) has served as a non-invasive diagnostic method for cystic fibrosis (CF) a disease of chloride channel expression and function in secretory epithelia. Investigators have also used NPD to demonstrate ion transport abnormalities in newborns with respiratory distress. Standard perfusates for diagnostic NPD studies include the use of amiloride, replacement of chloride with gluconate, cAMP agonists, and nucleotides such as ATP. The pH of these perfusates may also be relevant to NPD studies as we have previously shown that the respiratory epithelia in mammals express CLC-2, which is a pH sensitive chloride channel. We hypothesized that acidic pH might activate chloride secretion in vivo if CLC-2 is present in human respiratory epithelia. Our objective was to determine the effect of acidic pH on NPD measurements and the frequency of expression of CLC-2 in normal subjects. Healthy adults were recruited and CLC-2 protein expression was detected in 20 of 29 primary nasal epithelial cell cultures. Acidic pH stimulated NPD responses in 33% of subjects. These findings suggest that pH sensitive alternative pathways are available for modulation in human respiratory epithelia and that NPD protocols should standardize pH of perfusates. PMID- 16369926 TI - No role for kaposi sarcoma-associated herpesvirus in pediatric idiopathic pulmonary hypertension. AB - Idiopathic pulmonary hypertension (IPHT) is a rare but progressive disease with devastating outcomes, especially in children. The etiology is unknown. A recent study reported a potential role of Kaposi sarcoma-associated herpesvirus in the pathogenesis of IPHT in adults. Our goal was to test if this association exists in pediatric patients. The pathology specimen database of the Children's Hospital of Pittsburgh from 1980-2004 was searched, and all patients with a diagnosis of IPHT were enrolled. Lung tissues containing the characteristic plexiform lesion from all patients were selected and stained with antibody to latent nuclear antigen (LNA-1) of Kaposi sarcoma-associated herpesvirus. To avoid false-negative results, three different positive controls were utilized. Eighteen patients were identified; 3 had familial pulmonary hypertension, and 2 had persistent pulmonary hypertension of infancy. Patients' ages varied from 3 days to 17 years, and the female-to-male ratio was 2:1. No positive history of HIV or of parents with HIV was identified. The Heath-Edwards grade of pulmonary hypertension was between 3 5. All patients died: 11 from the disease, and the remaining 7 died after receiving a lung transplant. Immunohistochemical staining with LNA-1 antibody showed no staining of either endothelial or smooth muscle cell nuclei in any of the patients' lung tissues, while all three positive control specimens showed the characteristic intranuclear granular, punctate staining pattern. Our results do not support a role for Kaposi sarcoma-associated herpesvirus in the pathogenesis of pediatric IPHT. PMID- 16369929 TI - Characterization of a novel polymorphic form of celecoxib. AB - A new solid form (Form IV) of celecoxib was prepared in the presence of Polysorbate 80 and HPMC. A celecoxib suspension containing the Form IV had significantly higher bioavailability (>4 times) in dogs than the marketed capsules and the suspension containing bulk drug powders (Form III). The new form was characterized using differential scanning calorimetry, powder X-ray diffraction (PXRD), scanning electron microscopy (SEM), infrared spectroscopy, and Raman spectroscopy. The solids separated from the suspension containing the new form showed a melt onset at 145-148 degrees C, which was about 12-15 degrees less than known melting points of Form I, II ,and III of celecoxib. The PXRD pattern of the separated solids was not consistent with any of the known celecoxib crystal forms or the known excipients in the suspension. The formation of the new solid form (Form IV) was dependent upon the concentration and ratio of HPMC and Polysorbate 80. A faster dissolution rate (>2 times) of Form IV was observed compared to the thermodynamically stable form of celecoxib (Form III). There were no measurable changes in the solid state of Form IV either in dried solids or in the suspension for at least 6 months at 40 degrees C and 16 months at 25 degrees C. PMID- 16369928 TI - Carrier-mediated hepatic uptake of a novel nonrenal excretion type uric acid generation inhibitor, Y-700. AB - Y-700, a novel xanthine oxidase inhibitor, was recently developed for the treatment of hyperuricemia and gout. Since the major elimination route of this compound is hepatic metabolism and excretion, the aim of the present study was to characterize the uptake mechanism of Y-700 in the liver, which is also the pharmacological target of Y-700. Efficient uptake of Y-700 was observed both in the liver in vivo and in isolated rat hepatocytes. The uptake was Na(+) dependent, saturable and inhibited both by ATP-depressants and various organic anions. Indomethacin competitively inhibited Y-700 uptake, whereas the inhibitory effect of organic cations and nucleosides was not so remarkable. Saturable and Na(+)-dependent uptake of Y-700 was also observed in freshly isolated human hepatocytes. Uptake of Y-700 by sinusoidal membrane transporters, such as organic anion transporter (Oat) 2 and organic anion transporting polypeptide (OATP)-B, OATP-C, OATP-8, and Oatp1, could not be detected although uptake of Y-700 in the oocytes expressing sodium/taurocholate cotransporting polypeptide (NTCP) was slightly observed. In conclusion, active transport system(s), which specifically recognize certain types of anionic compounds, are involved in the hepatic uptake of Y-700 and, at least partially, relevant to its elimination from the circulation as well as delivery to pharmacological target. PMID- 16369930 TI - Contribution of mass spectrometry to assess quality of milk-based products. AB - The vast knowledge of milk chemistry has been extensively used by the dairy manufacturing industry to develop and optimize the modern technology required to produce high-quality milk products to which we are accustomed. A thorough understanding of the chemistry of milk and its numerous components is essential for designing processing equipment and conditions needed for the manufacture and distribution of high-quality dairy products. Knowledge and application of milk chemistry is also indispensable for fractionating milk into its principal components for use as functional and nutritional ingredients by the food industry. For all these reasons, powerful analytical methods are required. Because of the complexity of the milk matrix, mass spectrometry, coupled or not to separation techniques, constitutes a key tool in this area. In the present manuscript, we review the contribution and potentialities of mass spectrometry based techniques to assess quality of milk-based products. PMID- 16369931 TI - Human amyloid beta-induced neuroinflammation is an early event in neurodegeneration. AB - Using a human amyloid beta (Abeta) intracerebroventricular infusion mouse model of Alzheimer's disease-related injury, we previously demonstrated that systemic administration of a glial activation inhibitor could suppress neuroinflammation, prevent synaptic damage, and attenuate hippocampal-dependent behavioral deficits. We report that Abeta-induced neuroinflammation is an early event associated with onset and progression of pathophysiology, can be suppressed by the glial inhibitor over a range of intervention start times, and is amenable to suppression without inhibiting peripheral tissue inflammatory responses. Specifically, hippocampal neuroinflammation and neurodegeneration occur in close time proximity at 4-6 weeks after the start of infusion. Intraperitoneal administration of inhibitor for 2-week intervals starting at various times after initiation of Abeta infusion suppresses progression of pathophysiology. The glial inhibitor is a selective suppressor of neuroinflammation, in that it does not block peripheral tissue production of proinflammatory cytokines or markers of B- and T-cell activation after a systemic lipopolysaccharide challenge. These results support a causal link between neuroinflammation and neurodegeneration, have important implications for future therapeutic development, and provide insight into the relative time window for targeting neuroinflammation with positive neurological outcomes. PMID- 16369932 TI - NG2 upregulation in the denervated rat fascia dentata following unilateral entorhinal cortex lesion. AB - The chondroitin sulfate proteoglycan NG2 is a component of the glial scar following brain injury. Because of its growth inhibiting properties, it has been suggested to impede axonal regeneration. To study whether NG2 could also regulate axonal growth in denervated brain areas, changes in NG2 were studied in the rat fascia dentata following entorhinal deafferentation and were correlated with the post-lesional sprouting response. Laser microdissection was employed to selectively harvest the denervated molecular layer and combined with quantitative RT-PCR to measure changes in NG2 mRNA (6 h, 12 h, 2 days, 4 days, 7 days post lesion). This revealed increases of NG2 mRNA at day 2 (2.5-fold) and day 4 (2 fold) post-lesion. Immunocytochemistry was used to detect changes in NG2 protein (1 days, 4 days, 7 days, 10 days, 14 days, 30 days, 6 months post-lesion). NG2 staining was increased in the denervated outer molecular layer at day 1 post lesion, reached a maximum 10 days post-lesion, and returned to control levels thereafter. Electron microscopy revealed NG2 immunoprecipitate on glial surfaces and in the extracellular matrix around neuronal profiles, indicating that NG2 is secreted following denervation. Double labeling of NG2-immunopositive cells with markers for astrocytes, microglia/macrophages, and mature oligodendrocytes suggested that NG2 cells are a distinct glial subpopulation before and after entorhinal deafferentation. BrdU labeling revealed that some of the NG2-positive cells are generated post-lesion. Taken together, our data revealed a layer specific upregulation of NG2 in the denervated fascia dentata that coincides with the sprouting response. This suggests that NG2 could regulate lesion-induced axonal growth in denervated areas of the brain. PMID- 16369933 TI - Gene profiling and bioinformatic analysis of Schwann cell embryonic development and myelination. AB - To elucidate the molecular mechanisms involved in Schwann cell development, we profiled gene expression in the developing and injured rat sciatic nerve. The genes that showed significant changes in expression in developing and dedifferentiated nerve were validated with RT-PCR, in situ hybridisation, Western blot and immunofluorescence. A comprehensive approach to annotating micro-array probes and their associated transcripts was performed using Biopendium, a database of sequence and structural annotation. This approach significantly increased the number of genes for which a functional insight could be found. The analysis implicates agrin and two members of the collapsin response-mediated protein (CRMP) family in the switch from precursors to Schwann cells, and synuclein-1 and alphaB-crystallin in peripheral nerve myelination. We also identified a group of genes typically related to chondrogenesis and cartilage/bone development, including type II collagen, that were expressed in a manner similar to that of myelin-associated genes. The comprehensive function annotation also identified, among the genes regulated during nerve development or after nerve injury, proteins belonging to high-interest families, such as cytokines and kinases, and should therefore provide a uniquely valuable resource for future research. PMID- 16369934 TI - Functional expression of Kir4.1 channels in spinal cord astrocytes. AB - Spinal cord astrocytes (SCA) have a high permeability to K+ and hence have hyperpolarized resting membrane potentials. The underlying K+ channels are believed to participate in the uptake of neuronally released K+. These K+ channels have been studied extensively with regard to their biophysics and pharmacology, but their molecular identity in spinal cord is currently unknown. Using a combination of approaches, we demonstrate that channels composed of the Kir4.1 subunit are responsible for mediating the resting K+ conductance in SCA. Biophysical analysis demonstrates astrocytic Kir currents as weakly rectifying, potentiated by increasing [K+]o, and inhibited by micromolar concentrations of Ba2+. These currents were insensitive to tolbutemide, a selective blocker of Kir6.x channels, and to tertiapin, a blocker for Kir1.1 and Kir3.1/3.4 channels. PCR and Western blot analysis show prominent expression of Kir4.1 in SCA, and immunocytochemistry shows localization Kir4.1 channels to the plasma membrane. Kir4.1 protein levels show a developmental upregulation in vivo that parallels an increase in currents recorded over the same time period. Kir4.1 is highly expressed throughout most areas of the gray matter in spinal cord in vivo and recordings from spinal cord slices show prominent Kir currents. Electrophysiological recordings comparing SCA of wild-type mice with those of homozygote Kir4.1 knockout mice confirm a complete and selective absence of Kir channels in the knockout mice, suggesting that Kir4.1 is the principle channel mediating the resting K+ conductance in SCA in vitro and in situ. PMID- 16369935 TI - Individualizing combination of two antiproliferative immunosuppressants with pharmacodynamic modeling of stimulated lymphocyte responses. AB - BACKGROUND: Unpredictable serious adverse events (SAE) of immunosuppression, e.g. nephrotoxicity, with the nephrotoxic immunosuppressants have fostered interest in alternative regimens, which contain two antiproliferative agents, and individualized therapy. However, titration of such combinations to individual needs is not understood. SPECIFIC AIM: To determine concentration (C) mixtures of mycophenolate mofetil (MMF) and sirolimus (SRL), which produce half-maximal inhibitory effect (EC(50)) on human lymphocytes from individual subjects. METHODS: Concentration mixtures of MMF (0-5 mug/ml) and SRL (0-30 ng/ml) were incubated with whole blood from each of five healthy human subjects. The intracellular cytokines IL-2, TNF-alpha, and IFN-gamma were measured in PMA ionomycin-stimulated T-cells (CD4+), while CD54, CD95, CD86, CD25, CD69, and CD71 were measured in pokeweed mitogen-stimulated B-cells, by flow cytometry. Pharmacodynamic (PD) relationships were evaluated using Hill equations modified for single and multidrug regimens, and expressed as EC(50) for each target receptor. RESULTS: No change was seen in the expression of the T-cell cytokines with either MMF or SRL. Each B-cell receptor was inhibited with increasing concentrations of either MMF or SRL. Each B-cell receptor was also inhibited half maximally at lower concentrations of MMF in the presence of SRL, than with either agent alone, for the test population of five subjects together, and for each of five individual subjects. However, each subject showed distinctly different amounts of MMF and SRL that needed to be present together, in order to produce an identical inhibitory effect on lymphocyte function. CONCLUSIONS: PD analysis of biological effect can potentially predict optimal concentration mixtures of two immunosuppressants for individual recipients, and enhance rejection prophylaxis and safety. While this holds promise for drug development efforts, clinical application must await correlation of lymphocyte markers with post-transplant clinical outcomes. PMID- 16369936 TI - Electrospray mass spectrometry (ESI-MS) in the study of metal-ligand solution equilibria. AB - In the 20 years, since the introduction of electrospray mass spectrometry (ESI MS), the use of this technique in various fields of inorganic, organometallic, and analytical chemistry has been steadily increasing. In this study, the application of ESI-MS to the study of metal-ligand solution equilibria is reviewed (till 2004 included). In a first section, advantages and drawbacks of ESI-MS in this type of application are described. Subsequently, a list of ca. 300 studies is reported, in which ESI-MS was used to give number and stoichiometry of the species at equilibrium, or also to estimate their stability constants. All studies are classified according to the metal ions under examination. Other related applications, such as host-guest interactions and metal ion-protein binding studies, are briefly reviewed as well. PMID- 16369937 TI - The slow road to the eukaryotic genome. AB - The eukaryotic genome is a mosaic of eubacterial and archaeal genes in addition to those unique to itself. The mosaic may have arisen as the result of two prokaryotes merging their genomes, or from genes acquired from an endosymbiont of eubacterial origin. A third possibility is that the eukaryotic genome arose from successive events of lateral gene transfer over long periods of time. This theory does not exclude the endosymbiont, but questions whether it is necessary to explain the peculiar set of eukaryotic genes. We use phylogenetic studies and reconstructions of ancestral first appearances of genes on the prokaryotic phylogeny to assess evidence for the lateral gene transfer scenario. We find that phylogenies advanced to support fusion can also arise from a succession of lateral gene transfer events. Our reconstructions of ancestral first appearances of genes reveal that the various genes that make up the eukaryotic mosaic arose at different times and in diverse lineages on the prokaryotic tree, and were not available in a single lineage. Successive events of lateral gene transfer can explain the unusual mosaic structure of the eukaryotic genome, with its content linked to the immediate adaptive value of the genes its acquired. Progress in understanding eukaryotes may come from identifying ancestral features such as the eukaryotic splicesome that could explain why this lineage invaded, or created, the eukaryotic niche. PMID- 16369938 TI - Plants on red alert: do insects pay attention? AB - Two recent hypotheses have proposed that non-green plant colouration evolved as a defence against herbivores, either as protective colouration promoting handicap signals indicating plant fitness or by undermining their crypsis. The handicap hypothesis posits a co-evolutionary process between plants and herbivores, whereas the anti-crypsis hypothesis suggests that an arms race between insects and plants is the evolutionary mechanism. Both explanations assume that insects are the evolutionary origin causing plants' colouration. Here, we propose a different hypothesis, termed the "Defence Indication hypothesis". This idea focuses on the multiple protective functions of anthocyanins and carotenoids as pigments, and suggests that plant colouration evolved primarily in response to various stressors. Because pigments and defensive compounds share a common biosynthesis, the production of pigments also provides elevated defensive strengths against herbivores, a process termed priming. In effect, the Defence Indication hypothesis predicts that pleiotropic effects of the pigments and, more generally, plants' shared defence responses, explain why insects might react to plant colouration. PMID- 16369939 TI - Multicellular redox regulation: integrating organismal biology and redox chemistry. AB - Early in the 20th century, Charles Manning Child attributed organismal gradients in metabolism to interactions among groups of cells. Metabolic gradients are now firmly grounded in redox chemistry, yet modern work on metabolic signaling has consistently focused on the cellular level. Multicellular redox regulation, however, may occur when redox state is determined by the behavior of a group of cells. For instance, typically an abundance of substrate will shift the redox state of mitochondria in the direction of reduction, leading to increased reactive oxygen species (ROS). These ROS, in turn, may modify the conformation and activity of proteins involved in signaling pathways, resulting in phenotypic changes. In contrast, if substrate triggers the contractions of a muscular structure comprising mitochondrion-rich cells, the resulting metabolic demand may shift the redox state in the direction of oxidation, with a corresponding decrease of ROS and different phenotypic effects. Indeed, colonial hydroids exemplify this process. Parallel examples may occur whenever mitochondria are concentrated in cells of structures that can respond to environmental perturbations with increased metabolic demand. In these circumstances, predicting the direction of metabolic signaling may require an understanding of events at the organismal level. PMID- 16369940 TI - Hypothesis: transcript-templated repair of DNA double-strand breaks. AB - Two mechanisms are available for the repair of DNA double-strand breaks (DSBs) in eukaryotic cells: homology directed repair (HDR) and non-homologous end-joining (NHEJ). While NHEJ is not restricted to a particular phase of the cell cycle, it is incapable of accurately repairing DBSs that have suffered a loss or gain of nucleotide sequence information. In contrast, HDR achieves accurate repair of such DSBs by use of a sister chromatid as a DNA template, but is restricted to cell cycle phases (S/G2) when such templates are available. In this scheme, G1 cells appear to lack a mechanism for the accurate repair of certain DSBs, and an ability to use alternative templates would be highly advantageous. Considered here, therefore, is the possibility that RNA transcripts are used as templates for HDR. Potential mechanisms for transcript-templated HDR, and ways in which it might be detected, are presented. PMID- 16369941 TI - And yet another epicycle. PMID- 16369943 TI - Controlling the stem cell niche: right time, right place, right strength. AB - Wnt signalling through beta-catenin plays a pivotal role during embryonic pattern formation, cell fate determination and tissue homeostasis in the adult organism. In the skin, as in many other tissues, Wnt/beta-catenin signalling can control lineage determination and differentiation. However, it was not known whether Wnt/beta-catenin signalling is an immediate regulator of the stem cell niche in skin tissue. A recent publication now provides evidence that Wnt/beta-catenin signalling exerts a direct effect on the stem cell compartment by inducing quiescent stem cells to enter the cell cycle during early stages of hair follicle regeneration. In addition, the authors demonstrate that beta-catenin is required for maintenance of the stem cell pool in the tissue. The data suggest that a gradient in Wnt/beta-catenin activity levels can induce different responses within distinct cell populations reflected by activation of distinct transcriptional profiles. PMID- 16369944 TI - Resistance and the jumping gene. AB - Transposons are well-known architects of genetic change but their role in insecticide resistance has, until recently, only been speculated upon. Transposon insertion, or transposon-mediated transposition, could alter either metabolic enzymes capable of degrading pesticides or could change the functionality of insecticide targets. The recent work of Aminetzach and coworkers suggests an exciting alternative, that transposon insertion can cause resistance by altering gene product function. This hypothesis is discussed in the light of other examples in which transposons have been implicated in insecticide resistance. PMID- 16369945 TI - Signal transduction in bacterial chemotaxis. AB - Motile bacteria respond to environmental cues to move to more favorable locations. The components of the chemotaxis signal transduction systems that mediate these responses are highly conserved among prokaryotes including both eubacterial and archael species. The best-studied system is that found in Escherichia coli. Attractant and repellant chemicals are sensed through their interactions with transmembrane chemoreceptor proteins that are localized in multimeric assemblies at one or both cell poles together with a histidine protein kinase, CheA, an SH3-like adaptor protein, CheW, and a phosphoprotein phosphatase, CheZ. These multimeric protein assemblies act to control the level of phosphorylation of a response regulator, CheY, which dictates flagellar motion. Bacterial chemotaxis is one of the most-understood signal transduction systems, and many biochemical and structural details of this system have been elucidated. This is an exciting field of study because the depth of knowledge now allows the detailed molecular mechanisms of transmembrane signaling and signal processing to be investigated. PMID- 16369946 TI - Molecular and cellular organization of insect chemosensory neurons. AB - Animals use their chemosensory systems to detect and discriminate among chemical cues in the environment. Remarkable progress has recently been made in our knowledge of the molecular and cellular basis of chemosensory perception in insects, based largely on studies in Drosophila. This progress has been possible due to the identification of gene families for olfactory and gustatory receptors, the use of electro-physiological recording techniques on sensory neurons, the multitude of genetic manipulations that are available in this species, and insights from several insect model systems. Recent studies show that the superfamily of chemoreceptor proteins represent the essential elements in chemosensory coding, endowing chemosensory neurons with their abilities to respond to specific sets of odorants, tastants or pheromones. Investigating how insects detect chemicals in their environment can show us how receptor protein structures relate to ligand binding, how nervous systems process complex information, and how chemosensory systems and genes evolve. PMID- 16369947 TI - X-linked imprinting: effects on brain and behaviour. AB - Imprinted genes are monoallelically expressed in a parent-of-origin-dependent manner and can affect brain and behavioural phenotypes. The X chromosome is enriched for genes affecting neurodevelopment and is donated asymmetrically to male and female progeny. Hence, X-linked imprinted genes could potentially influence sexually dimorphic neurobiology. Consequently, investigations into such loci may provide new insights into the biological basis of behavioural differences between the sexes and into why men and women show different vulnerabilities to certain mental disorders. In this review, we summarise recent advances in our knowledge of X-linked imprinted genes and the brain substrates that they may act upon. In addition, we suggest strategies for identifying novel X-linked imprinted genes and their downstream effects and discuss evolutionary theories regarding the origin and maintenance of X-linked imprinting. PMID- 16369948 TI - Molecular features of meiotic recombination hot spots. AB - Meiotic recombination occurs preferentially at certain regions called hot spots and is important for generating genetic diversity and proper segregation of chromosomes during meiosis. Hot spots have been characterized most extensively in yeast, mice and humans. The development of methods based on sperm typing and population genetics has facilitated rapid and high-resolution mapping of hot spots in mice and humans in recent years. With increasing information becoming available on meiotic recombination in different species, it is now possible to compare several molecular features associated with hot-spot loci. Further, there have been advances in our knowledge of the factors influencing hot-spot activity and the role that they play in structuring the genome into haplotype blocks. We review the molecular features associated with hot spots in terms of their properties and mechanisms underlying their function and distribution. A large number of these features seem to be shared among hot spots from different species suggesting common mechanisms for their formation and function. PMID- 16369949 TI - Use of bacteria in anti-cancer therapies. AB - While a number of valid molecular targets have been discovered for tumours over the past decade, finding an effective way of delivering therapeutic genes specifically to tumours has proved more problematic. A variety of viral and non viral delivery vehicles have been developed and applied in anti-cancer gene therapies. However, these suffer from either inefficient and/or short-lived gene transfer to target cells, instability in the bloodstream and inadequate tumour targeting. Recently, various types of non-pathogenic obligate anaerobic and facultative anaerobic bacteria have been shown to infiltrate and selectively replicate within solid tumours when delivered systemically. This has prompted the development of cancer gene therapy protocols that use such bacteria as gene delivery vehicles. Here, we review the evidence for the success of these in pre clinical models and clinical trials, as single modality treatments and in combination with conventional cancer therapies. PMID- 16369950 TI - What is new about ADP-ribosylation? PMID- 16369951 TI - Cancer cell undifferentiation: a matter of expression rather than mutations? PMID- 16369952 TI - Interactive effects of cadmium and all-trans-retinoic acid on the induction of forelimb ectrodactyly in C57BL/6 mice. AB - BACKGROUND: Most toxicological studies have tested single chemical agents at relatively high doses, and fewer studies have addressed the toxic effects of chemical interactions. It is important to understand the toxicity of chemical mixtures in order to assess the more realistic risks of environmental and occupational exposures. A number of chemicals are known to induce a predominantly postaxial forelimb ectrodactyly in C57BL/6 mice, including acetazolamide, ethanol, cadmium, valproic acid, carbon dioxide, dimethadione, phenytoin, and 13 cis-retinoic acid and all-trans-retinoic acid (RA). In the present study, the interactive effects of coadministration of cadmium and RA on developing limbs were investigated. METHODS: Pregnant C57BL/6 mice were treated with different intraperitoneal (IP) doses of cadmium chloride (CdCl2) and/or RA on gestational day (GD) 9.5, and fetuses were collected on GD 18 and double stained for examination of skeletal defects. RESULTS: When RA was given simultaneously with cadmium, a significant increase in the incidence and severity of forelimb ectrodactyly (predominantly postaxial) was observed compared to the results with corresponding doses of cadmium or RA alone. When mice were exposed to subthreshold doses of both cadmium (0.5 mg/kg) and RA (1 mg/kg), the combined treatment exceeded the threshold, resulting in forelimb ectrodactyly in 19% of the fetuses. Moreover, coadministration of cadmium and RA at doses exceeding the respective thresholds showed a synergistic effect, that is, 92% of fetuses were found with the forelimb defect as opposed to 10% if the response were additive. CONCLUSIONS: The findings demonstrate that concurrent exposure to these teratogens can have a synergistic effect and that subteratogenic doses may combine to exceed a threshold. PMID- 16369953 TI - Isolated extramedullary relapse after autologous bone marrow transplantation for acute myeloid leukemia: case report and review of the literature. AB - Isolated extramedullary relapse (IEMR) is a pattern of acute myeloid leukemia (AML) relapse post-allogeneic bone marrow transplantation (alloBMT). Less is known about IEMR post-autologous BMT (autoBMT) and about factors associated with IEMR. We report a case of a woman with M4 AML who experienced IEMR post-autoBMT and review the related literature. Seventy-two alloBMT and 3 autoBMT patients, including ours, were identified. The review suggests that an M2 or M4 French American-British (FAB) phenotype, intermediate cytogenetic risk group, and chromosome 8 abnormalities are more frequently associated with the occurrence of IEMR. IEMR occurs earlier in autoBMT than in alloBMT. Combined treatment with radiation and high-dose chemotherapy may be effective. When we searched the European Bone Marrow Transplant Registry (EBMTR) database, we found the incidence of IEMR to be statistically greater in alloBMT than in autoBMT (11% vs. 6%; P = 0.02), but no correlations have been found with the conditioning transplant regimen used. A closer follow-up, including body and central nervous system scan, should be considered in patients who are undergoing BMT presenting with several IEMR-associated factors. PMID- 16369954 TI - Reticulocyte evaluation in alpha(+)-thalassemia. AB - Although it is almost certain that alpha(+)-thalassemia protects against malaria, the mechanisms for that are still unknown. It has been suggested that an increased number of young circulating red blood cells in alpha(+)-thalassemic children, as a result of some degree of ineffective erythropoiesis, could be related to the high frequencies of the alpha(+)-thalassemic allele in malaria endemic areas. Reticulocyte evaluation in this condition, however, has been poorly performed so far. Our objective was to determine the reticulocyte number and maturation degree, in addition to the soluble transferrin receptor and serum erythropoietin levels, in alpha(+)-thalassemia heterozygotes, comparing them with normal alpha-genotype controls. One hundred twenty-one alpha(+)-thalassemia carriers (-alpha(3.7)/alphaalpha) and 249 controls (alphaalpha/alphaalpha), all of them with normal serum ferritin levels, were subclassified according to age (1 5, 6-10, 11-15, 16-20, and over 20 years old). Reticulocyte analyzes were carried out by flow cytometry and sTfR and s-Epo levels determined by immunonephelometry and chemiluminescence, respectively. The comparisons did not show any significant difference between thalassemics and controls regarding the reticulocyte parameters [percentages and absolute values, P = 0.2643 and 0.5421; high, medium, and low maturation degree, P = 0.2579, 0.2196, and 0.4192; RET maturity index (RMI), P = 0.2471, respectively], as well as the s-Epo levels (P = 0.5711). The sTfR concentrations were higher in the thalassemic group (P = 0.0001), but statistical significance was due only to the 1-5 and over 20 subgroups (P = 0.0082 and 0.0436, respectively). The results found here are compatible with a compensated erythropoiesis and do not confirm the hypothesis mentioned above. PMID- 16369955 TI - Granulocytic sarcoma of the gingiva with trisomy 21. PMID- 16369957 TI - Federal repatriation legislation and the role of physical anthropology in repatriation. AB - Two laws governing the disposition of Native American human remains in museums and institutions have had a profound impact on anthropology, and especially physical anthropology. In contrast to the perception of constant conflict between Native Americans and physical anthropologists, the repatriation process based on these laws has been in large part harmonious between institutions and Native peoples in the US. Despite misconceptions, the Native American Graves Protection and Repatriation Act (NAPGRA; 25 United States Code (U.S.C.) 3001-3013) was not intended to halt further research on Native American remains in museums. In fact, court decisions have affirmed that the documentation of human remains produces information no other methods can provide, and provides necessary evidence to be incorporated and weighed, along with other evidence, in evaluating "cultural affiliation," the legal term for the required connection from federally recognized Native American groups to their ancestors. The wide variety of osteological data collected at the National Museum of Natural History (NMNH), Smithsonian Institution, have proven indispensable when evaluating cultural affiliation, especially when other information sources are unhelpful or ambiguous, and provide an empirical basis for determining the ancestry of individuals whose remains will be discovered in the future. To date, the claim driven process at the NMNH has resulted in the affiliation and repatriation of more Native American remains than any other institution in the country. Repatriation experiences at the NMNH demonstrate the changing relationships between museums and Native peoples, the continuing important contributions that physical anthropology makes to the repatriation process, and the importance of physical anthropology in understanding the recent and ancient history of North America. PMID- 16369958 TI - Deep time and the search for anthropoid origins. AB - Recent fossil discoveries, phylogenetic analyses, revised reconstructions of continental drift, and accumulating molecular evidence have all yielded new information relating to anthropoid origins within the broader context of primate evolution. There is an emerging consensus among molecular studies that four superorders of eutherian mammals can be recognized: Afrotheria, Euarchontoglires (to which primates belong), Laurasiatheria, and Xenarthra. Overall, molecular phylogenies for mammals agree with some statistical analyses of the primate fossil record in indicating an early origin for primates around 85 Ma ago, and the divergence of haplorhines and strepsirrhines at ca. 77 Ma. Such an ancient date for the origin of haplorhines is some 17 Ma prior to the first known possible primate, and some 22 Ma before the earliest fossil evidence of undoubted euprimates. Because anthropoid fossils date back at least to the late Eocene and perhaps to the middle Eocene, and given indications of an early origin for primates, it is unlikely that ancestral anthropoids arose within any other currently known clade of fossil primates (adapiforms, omomyiforms, strepsirrhines, or tarsiiforms). Implications of new molecular, morphological, and biogeographic lines of evidence are explored with respect to the likely time and place of the origin of anthropoids. Four competing, testable hypotheses are reviewed in detail: 1) the Paratethyan hypothesis, 2) the continental Asian hypothesis, 3) the Indo-Madagascar hypothesis, and 4) the African hypothesis. A case is made that current evidence best supports a relatively ancient Gondwanan origin for primates, as well as a Gondwanan (African or Indo-Madagascan) origin for anthropoids at least as old as that of any other currently documented major primate clade. Available fossil evidence at present seems to be most compatible with the African hypothesis, but it is noteworthy that primates are included not in Afrotheria but in Euarchontoglires. PMID- 16369959 TI - Modeling physiological and anthropometric variables known to vary with body size and other confounding variables. AB - This review explores the most appropriate methods of identifying population differences in physiological and anthropometric variables known to differ with body size and other confounding variables. We shall provide an overview of such problems from a historical point of view. We shall then give some guidelines as to the choice of body-size covariates as well as other confounding variables, and show how these might be incorporated into the model, depending on the physiological dependent variable and the nature of the population being studied. We shall also recommend appropriate goodness-of-fit statistics that will enable researchers to confirm the most appropriate choice of model, including, for example, how to compare proportional allometric models with the equivalent linear or additive polynomial models. We shall also discuss alternative body-size scaling variables (height, fat-free mass, body surface area, and projected area of skeletal bone), and whether empirical vs. theoretical scaling methodologies should be reported. We shall offer some cautionary advice (limitations) when interpreting the parameters obtained when fitting proportional power function or allometric models, due to the fact that human physiques are not geometrically similar to each other. In conclusion, a variety of different models will be identified to describe physiological and anthropometric variables known to vary with body size and other confounding variables. These include simple ratio standards (e.g., per body mass ratios), linear and additive polynomial models, and proportional allometric or power function models. Proportional allometric models are shown to be superior to either simple ratio standards or linear and additive polynomial models for a variety of different reasons. These include: 1) providing biologically interpretable models that yield sensible estimates within and beyond the range of data; and 2) providing a superior fit based on the Akaike information criterion (AIC), Bayes information criterion (BIC), or maximum log likelihood criteria (resulting in a smaller error variance). As such, these models will also: 3) naturally lead to a more powerful analysis-of-covariance test of significance, which will 4) subsequently lead to more correct conclusions when investigating population (epidemiological) or experimental differences in physiological and anthropometric variables known to vary with body size. PMID- 16369960 TI - Role of physical anthropology in intermediate and secondary education. AB - The role of physical anthropology in precollegiate education has been limited, but has the potential to play a vital and integral role in promoting students' understanding of themselves and others. This study describes the development, implementation, and results of a program constructed on an inquiry-based learning model that introduces middle and high school students to the content of physical anthropology. Demonstrated student learning indicates that intermediate and secondary students are capable of acquiring and applying complex content about human evolution and diversity. Program findings indicate that students frequently had previous knowledge about the hominid fossil record but little information about the significance of human biological diversity. Teaching intermediate and high school students about our common evolutionary heritage and the fact that humans exhibit clinal variation were found to be both powerful and effective content material. The fact that program participants lacked previous knowledge about the relationship between human biological diversity (illustrated in this program by light and dark skin color) and the erroneous sociocultural construction of this diversity confirmed the suspicion that this content is not being addressed in precollegiate education. Traditionally, intermediate and secondary students are taught about human variation within the context of social studies, reading, communication, and fine-arts classes. This program identifies a new paradigm for teaching about human variation. Providing students with scientific knowledge about human origins and diversity provides a constructive starting point, creating a common platform and knowledge base on which to then frame discussions about cultural variation. PMID- 16369961 TI - Haplotype trees and modern human origins. AB - A haplotype is a multisite haploid genotype at two or more polymorphic sites on the same chromosome in a defined DNA region. An evolutionary tree of the haplotypes can be estimated if the DNA region had little to no recombination. Haplotype trees can be used to reconstruct past human gene-flow patterns and historical events, but any single tree captures only a small portion of evolutionary history, and is subject to error. A fuller view of human evolution requires multiple DNA regions, and errors can be minimized by cross-validating inferences across loci. An analysis of 25 DNA regions reveals an out-of-Africa expansion event at 1.9 million years ago. Gene flow with isolation by distance was established between African and Eurasian populations by about 1.5 million years ago, with no detectable interruptions since. A second out-of-Africa expansion occurred about 700,000 years ago, and involved interbreeding with at least some Eurasian populations. A third out-of-Africa event occurred around 100,000 years ago, and was also characterized by interbreeding, with the hypothesis of a total Eurasian replacement strongly rejected (P < 10(-17)). This does not preclude the possibility that some Eurasian populations could have been replaced, and the status of Neanderthals is indecisive. Demographic inferences from haplotype trees have been inconsistent, so few definitive conclusions can be made at this time. Haplotype trees from human parasites offer additional insights into human evolution and raise the possibility of an Asian isolate of humanity, but once again not in a definitive fashion. Haplotype trees can also indicate which genes were subject to positive selection in the lineage leading to modern humans. Genetics provides many insights into human evolution, but those insights need to be integrated with fossil and archaeological data to yield a fuller picture of the origin of modern humans. PMID- 16369962 TI - Are modern environments really bad for us?: revisiting the demographic and epidemiologic transitions. AB - It is a common assumption that agriculture and modernization have been detrimental for human health. The theoretical argument is that humans are adapted to hunter-gatherer lifestyles, and that the agricultural and "modern" environments are novel and hence likely to be detrimental. In particular, changes in nutrition, and population size and distribution with the adoption of agriculture, are considered to increase the risk of infectious disease mortality. Similarly, changes due to modern lifestyles, notably changes in nutrition, smoking, exercise, and stress, are thought to be associated with an increased risk of degenerative disease mortality in the industrial environment. This paper reviews the available literature on the history and prehistory of total mortality (the demographic transition) and cause of death (the epidemiologic transition), and finds that neither agriculture nor modernization is associated with increases in mortality, i.e., declines in health. First, mortality does not appear to have increased during the transition to agriculture, or during the early phases of the industrial revolution. Clearly, infectious diseases have declined with modernization. Second, the empirical data, when uncorrected for misclassification of cause of death, do suggest an increase in degenerative disease mortality, at least until the mid 20th century, when these causes of death clearly began to decline. All studies that correct for misclassification of cause of death, however, find that the general decline in degenerative disease mortality began much earlier, perhaps as early as the 1850s in the developed countries. This is about the same time that infectious disease mortality began to decline in these countries. The exception is neoplasms, which increased with modernization until quite recently. Part of the increase in neoplasms may be attributable to increases in smoking during the course of modernization. Nevertheless, the overall risk of degenerative disease mortality appears to have declined with modernization. The fact that the decline in the risk of infectious disease mortality, and the decline in risk of degenerative disease mortality, are largely coordinated suggests that the causes of both declines may be related. Historical trends in morbidity, and potential causes of the decline in infectious and degenerative disease mortality, are briefly considered. PMID- 16369963 TI - Albinism (OCA2) in Amerindians. AB - Homozygosity for a mutation in the P locus mapped to the human chromosome 15q11.2 12 results in tyrosinase-positive albinism (OCA2). This type of albinism has a worldwide distribution, with a prevalence of about 1 in 36,000 among European Americans in the United States. It has a moderate to relatively high prevalence values (1 in 28 to 1 in 6,500) in various Amerindian populations in the southwestern United States, southern Mexico, eastern Panama, and southern Brazil. The wide distribution of the gene for OCA2 in Amerindian populations, and its relatively high frequency in several of these populations, are enigmatic because of the detrimental nature of OCA2 in the presence of certain environmental conditions. The relative Darwinian fitness of individuals with this inborn error of metabolism would have been reduced in early nomadic hunting-gathering populations because of their poor visual acuity and sensitivity to the sun. Nevertheless, specific situations allowed OCA2 to increase in frequency in certain Amerindian populations at various different times in history. The present objectives are to review the literature on albinism (OCA2) in Amerindians, and propose hypotheses for the variable frequencies of the OCA2 gene in Amerindian populations, which include chance processes (founder effect, bottleneck effect, and genetic drift) in small populations, natural selection, cultural selection, and the interaction of situations that led to the increase of the frequency of the albino gene in some generations. Special emphasis is placed on those Amerindian populations with a relatively high frequency of the OCA2 gene that have been best-studied, namely, the Cuna population of eastern Panama and the Hopi population in the southwestern US. Hypothetical scripts are presented for the present relatively high frequencies of the OCA2 gene in these populations. A hypothetical script is also presented, showing how a mutant gene could have reached a relatively high frequency in a small endogamous early Mayan population and then been spread by migrating groups to other geographical regions, following the rapid increase in size of that population. Comprehensive molecular studies of OCA2 genes in Amerindian populations could yield information on the possible origin of the albino gene present in many of these populations, in addition to the gene flow that occurred among some of them in past generations. The results of these studies could lead to more informed hypotheses concerning the wide distribution of OCA2 in Amerindian populations. PMID- 16369964 TI - Juan Comas's summary history of the American association of physical anthropologists (1928-1968). AB - This translation of Juan Comas's Summary History of the American Association of Physical Anthropologists was originally published in Spanish by the Instituto Nacional de Antropologia e Historia, Mexico, in 1969 (Departamento de Investigaciones Antropologicas, Publication 22). Physical anthropologists from North America and members of the American Association of Physical Anthropologists owe Juan Comas a debt of gratitude for having labored to produce this Summary History of the AAPA. There is much useful and interesting material in this document: extensive endnotes that are helpful to the historian of the profession; an appendix of the Journal issues where the proceedings of annual meetings can be found; a detailed listing of contributors of papers to annual meetings from 1930 1968; a warm acknowledgment and history of the contributions of the Wenner-Gren Foundation to biological anthropology; a history of the Yearbook of Physical Anthropology; and comments on the short-lived PA Newsletter. In addition, there are appendices with the founding AAPA Constitution and By-Laws from 1930 and as they existed in 1968. All of this synoptic information saves the reader with interests in the history of the AAPA considerable effort, especially when few university and college libraries have the full (old and new) series of the AJPA on their shelves. We have tried to provide a translation of Comas's history that is faithful to the original Spanish-language publication. In a few cases, we shortened sentences and applied a slightly more modern usage than was popular in the late 1960s. PMID- 16369965 TI - Therapeutic decision-making in inhibitor patients. AB - A panel of hematologists met to discuss treatment choices in patients with hemophilia A and inhibitors. The group agreed that the management of bleeding episodes is complicated by a variable response to bypassing therapy, uncertainties regarding optimal dosing of NovoSeven, and concerns about the combined use of NovoSeven and FEIBA. Thrombotic risk was rated very low for both agents, and there was consensus that product selection should not be restricted to recombinant products. Cost considerations in therapeutic decision-making were also discussed. PMID- 16369966 TI - Osteonecrosis of the jaw associated with pamidronate therapy. AB - Bisphosphonates are commonly used in the treatment and prevention of osteoporosis, and they are also an important therapeutic adjunct in multiple myeloma and other cancers metastatic to bone. Bisphosphonates are generally well tolerated and associated with minimal adverse effects; however, there exists a growing concern that intravenous bisphosphonate use is associated with osteonecrosis of the jaw (ONJ). We report the occurrence of osteonecrosis of the jaw associated with pamidronate therapy in 12 patients diagnosed with multiple myeloma, breast carcinoma, or renal cell carcinoma, all involving bone. At the onset of jaw osteonecrosis, pamidronate therapy was the single medication common to all 12 patients. The duration of therapy varied from 12 to 77 months before osteonecrosis was observed; 92% (11/12) of cases involved the posterior mandible and all cases have been refractory to a variety of medical therapies, including surgical debridement and systemic antibiotics. Available tissue biopsies revealed inflammation consistent with osteomyelitis. In one biopsy, Actinomyces spp. were recovered from culture, but treatment with an extended course of clindamycin conferred no clinical benefit. The persistence of exposed bone remains a significant source of morbidity and pain for each surviving patient. Discontinuation of pamidronate therapy has not helped reverse the presence of osteonecrosis, and surgical manipulation of the involved site appears to worsen the underlying bone pathology. ONJ is an important adverse outcome associated with bisphosphonate therapy, and physicians prescribing pamidronate or zoledronate must be aware of the association between these drugs and this serious clinical entity. Failure to recognize the signs of ONJ can lead to unnecessary surgical procedures, which ultimately exacerbate the condition and impact quality of life. The unremitting nature of this clinical development, and the long lasting morbidity associated with it suggests that patients should be counseled regarding the possible occurrence of ONJ prior to initiating therapy with pamidronate. PMID- 16369967 TI - Use of cefoperazone still needs a caution for bleeding from induced vitamin K deficiency. PMID- 16369968 TI - CD117 Expression as an aid to identify immature myeloid cells and foci of ALIP in bone marrow trephines. PMID- 16369969 TI - Prophylaxis of central venous catheter-related thrombosis with minidose warfarin in patients treated with high-dose chemotherapy and peripheral-blood stem-cell transplantation: retrospective analysis of 228 cancer patients. AB - Patients with a central venous catheter (CVC) undergoing high-dose chemotherapy (HDC) followed by peripheral-blood stem-cell transplantation (PBSCT) for malignancies are at high risk of thrombosis, but the use of anti-coagulant prophylaxis remains debatable in this setting of patients. We analyzed the efficacy and the safety of minidose warfarin in 228 patients in whom CVCs had been placed and who had received 292 HDC courses of therapy. The catheters remained in place for a mean of 173 (range 40-298) days. All patients received prophylactic oral warfarin in the fixed dose of 1 mg/day starting on the day of CVC insertion. Prophylaxis was interrupted during aplasia when platelet counts fell below 50,000/dL. There were no toxic deaths related to the prophylaxis. Overall there were 4 thrombotic events. Three occurrences were directly related to the catheter, while the remaining event was a deep saphenous-vein thrombosis. A number of potential predictive factors were analyzed for their impact on thrombotic events without finding any significant correlation. Four episodes of bleeding occurred, with each of these individuals having a normal INR but a platelet count below 50,000/dL. Minidose warfarin is effective and safe to use for preventing thrombotic events in this setting of patients. PMID- 16369970 TI - Therapy-related lymphomas in patients with autoimmune diseases after treatment with disease-modifying anti-rheumatic drugs. AB - Ten patients developing lymphomas after disease modifying anti-rheumatic drugs (DMARD) (methotrexate, n = 3, mean cumulative dose = 3.4 g; cyclophosphamide, n = 2, mean dose = 70 g; azathioprine, n = 6, mean dose = 243 g) were investigated. Methotrexate-related lymphomas were Epstein-Barr virus (EBV)-positive, had infrequent aberrant methylation of p15 and p16, and responded well to methotrexate withdrawal or anti-CD20 antibody (rituximab) alone without concomitant chemotherapy, implying that defective immunosurveillance was important in lymphomagenesis. However, 75% of cyclophosphamide/azathioprine related lymphomas were EBV-negative, had frequent p15 and p16 methylation, and responded poorly to drug withdrawal and chemotherapy, implying that direct drug induced mutagenesis might be involved in lymphomagenesis. PMID- 16369971 TI - Non-Hodgkin disease in beta-thalassemia major. AB - Thalassemia is a spectrum of diseases characterized by the decrease or absence of globin chains. The occurrence of lymphoma in thalassemia has rarely been reported, and our review of the English literature revealed only four cases. Because anemia is always masked by regular transfusions in thalassemic patients, physicians discover a hidden malignancy late in the course of the disease. We hereby report the case of a thalassemic patient developing non-Hodgkin disease and discuss the possibility of a link between the two disease entities. This case is intended to alert physicians of the possibility of a malignancy in thalassemia patients. PMID- 16369972 TI - Relative incidence of agranulocytosis and aplastic anemia. AB - Agranulocytosis and aplastic anemia are both rare, life-threatening blood dyscrasias. Agranulocytosis is mainly caused by medicines, whereas the etiology of aplastic anemia is largely unexplained. In two epidemiologic studies using the same methods, we observed a striking inverse relationship between the incidence of the two diseases in different regions, including five countries in Europe, and Israel and Thailand. The annual incidence of agranulocytosis ranged from 1.1 to 4.9 cases per million, and that of aplastic anemia, from 0.7 to 4.1 per million; the inverse correlation was consistent among the regions (R2 = 0.74). There is no clear explanation for this previously unreported pattern, but it seems unlikely to be due to methodology. PMID- 16369973 TI - Ikaros increases normal apoptosis in adult erythroid cells. AB - Ikaros is a critical transcriptional regulator of hematopoietic cell differentiation. In addition to its effects on the lymphoid system and hematopoietic stem-cell compartment, we have previously shown that Ikaros is also required for normal erythroid development. In this report, we compare Ikaros dependent gene expression in erythroid cells of mice lacking the Ikaros protein with that of normal mice in purified adult bone-marrow erythroid cells (BMRC). Gene expression, measured by Affymetrix microarray analysis, indicates that in the BMRC of Ikaros-null mice, there is significant up-regulation of SMADs 6 and 7, serine protease inhibitor 3, and immediate-early protein 3 (IER3), all proteins that play a modulating role in apoptosis. We investigate the role of Ikaros in oxidative stress-induced apoptosis using Annexin-V staining and FACS analysis. We find a decrease in apoptosis in the BMRC of Ikaros-null mice compared to normal mice. This effect is also seen in nonerythroid cells but is stronger in BMRC. We conclude that normal Ikaros function increases normal apoptosis in erythroid cells. The data also suggest that Ikaros plays a role in apoptosis-mediated events in other normal hematopoietic cell lineages. PMID- 16369974 TI - Exposure of blood from patients with sickle cell disease to air changes the morphological, oxygen-binding, and sickling properties of sickled erythrocytes. AB - We collected venous blood samples from 7 steady-state patients with homozygous sickle cell disease under venous oxygen pressure without exposure to air (UnExp blood) and compared the morphological, oxygen-binding, and sickling properties with those of SS cells in aliquots of the same venous blood samples that were oxygenated in room air or at a PO2 near 180 mmHg (Exp-blood). Results showed that (1) upon deoxygenation under nitrogen, UnExp-blood generated a significantly higher percentage of elongated reversibly sickled cells (RSCs) than did Exp blood; (2) upon gradual oxygenation of completely deoxygenated sickled cells, RSCs in UnExp-blood converted to discocytes at a higher oxygen pressure than did those in Exp-blood; (3) the degree of hysteresis between the sickling/desickling curves of UnExp-blood was greater than that of Exp-blood; and (4) deoxy-Hb S in hemolysate prepared from SS cells in UnExp-blood polymerized without a delay time, while those from Exp-blood polymerized with a distinct delay time. The in vivo properties of RSCs significantly changed upon oxygenation. We also found that the various properties of blood samples collected from patients with SCD by the ordinary method were similar to those of Exp-blood, probably because such blood samples are exposed to oxygen through air in the needle, syringe, and Vacutainer. Once SS cells were oxygenated, the in vivo properties of RSCs could not be recovered by partial deoxygenation to venous oxygen pressure. PMID- 16369975 TI - Granulocytic sarcoma giant in chronic myeloid leukemia during imatinib mesylate therapy. PMID- 16369976 TI - Success with infliximab in treating refractory hemophagocytic lymphohistiocytosis. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare disorder characterized by fever, pancytopenia, hepatosplenomegaly, liver dysfunction, and hemophagocytosis. A 29-year-old woman, diagnosed with systemic lupus erythematosus in 1996, developed HLH in early June 2002. HLH remained refractory during 1.5 months of treatment including corticosteroid, cyclosporine, plasma exchange, vincristine, and etoposide. Infliximab (5 mg/kg/day) was then administered twice. After the second administration, the patient attained remission. Because HLH itself is not a neoplasm but an uncontrolled immune reaction, blocking cytokines involved in the reaction should have therapeutic potentials. For HLH patients not responding to conventional therapy, anticytokine treatment with infliximab may represent one of promising options. PMID- 16369977 TI - Peripheral monocytes and CD4+ cells are potential sources for increased circulating levels of TGF-beta and substance P in autoimmune myelofibrosis. AB - Myelofibrosis is an uncommon phenomenon associated with a variety of neoplastic and inflammatory processes. Although there is evidence that cytokines elaborated by clonal malignant hematopoietic cells are implicated in myelofibrosis in primary hematologic disorders, there has been little data to date on the pathophysiology of myelofibrosis in autoimmune disorders. Here we report a case of autoimmune myelofibrosis with pancytopenia. Peripheral blood monocytes and CD4 positive lymphocytes produced significantly elevated levels of transforming growth factor beta (TGF-beta) compared to similar cells from healthy volunteer controls. TGF-beta has been implicated in the pathogenesis of myelofibrosis associated with primary hematological malignancies. Furthermore, substance P, previously linked to myelofibrosis, was also detected in elevated levels in the patient's serum and correlated negatively with the levels of the patient's blood counts. These findings suggest a role for both TGF-beta and substance P in the pathophysiology of autoimmune myelofibrosis. This is the first report of deregulated production of TGF-beta by monocytes in the pathobiology of autoimmune myelofibrosis. PMID- 16369978 TI - Primary lymphoma of the bladder: case report. PMID- 16369979 TI - Efficacy of mycophenolate mofetil as single-agent therapy for refractory immune thrombocytopenic purpura. AB - Refractory disease occurs in 25% or more of adults with idiopathic (immune) thrombocytopenic purpura (ITP). Therapy to elevate the platelet count may be required in a proportion of these patients. Immunosuppressive agents such as prednisone, azathioprine, cyclophosphamide, and cyclosporin have been shown to be effective treatments in a proportion of patients with refractory ITP. A newer immunosuppressive medication, mycophenolate mofetil (MMF), has been used successfully with acceptable toxicity in solid organ transplant patients to reduce the risk of organ rejection. The goal of this study was to determine whether MMF is an effective treatment for refractory ITP. Efficacy, defined as a sustained platelet increase to a level greater than 50 x 10(9)/L, was seen in 7 of 18 patients with refractory ITP. Three of these 7 patients have had intermittent thrombocytopenic episodes while continuing the medication. No severe toxicity was seen, although two of the 18 patients discontinued MMF within the first month of treatment because of side effects, i.e., headache. In summary, MMF may be a useful component of a combination protocol but does not appear to be highly effective as sole therapy in patients with refractory ITP. The data suggests that response rates to MMF may be higher in patients who have had a shorter duration of their ITP. PMID- 16369980 TI - Heparin-induced thrombocytopenia and thrombosis. AB - Heparin, employed clinically for more than 50 years, is still a widely used anticoagulant. Unfortunately, some patients given this agent develop thrombocytopenia and thrombosis. Because this side effect can have catastrophic consequences, it is imperative that all clinicians caring for patients who receive heparin have at least a basic understanding of its pathogenesis, diagnosis, and management. PMID- 16369981 TI - High-throughput process development for recombinant protein purification. AB - Methods development in chromatographic purification processes is a complex operation and has traditionally relied on trial and error approaches. The availability of a large number of commercial media, choice of different modes of chromatography, and diverse operating conditions contribute to the challenging task of accelerating methods development. In this paper, we describe a novel microtiter-plate based screening method to identify the appropriate sequence of chromatographic steps that result in high purities of bioproducts from their respective culture broths. Protein mixtures containing the bioproduct were loaded on aliquots of different chromatographic media in microtiter plates. Serial step elution of the proteins, in concert with bioproduct-specific assays, resulted in the identification of "active fractions" containing the bioproduct. The identification of a successful chromatographic step was based on the purity of the active fractions, which were then pooled and used as starting material for screening the next chromatographic dimension. This procedure was repeated across subsequent dimensions until single band purities of the protein were obtained. The sequence of chromatographic steps and the corresponding operating conditions identified from the screen were validated under scaled-up conditions. Various modes of chromatography including hydrophobic interaction, ion exchange (cation and anion exchange) and hydrophobic charge-induction chromatography (HCIC), and different operating conditions (pH, salt concentration and type, etc.) were employed in the screen. This approach was employed to determine the sequence of chromatographic steps for the purification of recombinant alpha-amylase from its cell-free culture broth. Recommendations from the screen resulted in single-band purity of the protein under scaled-up conditions. Similar results were observed for an scFv-beta-lactamase fusion protein. The use of a miniaturized screen enables the parallel screening of a wide variety of actual bioprocess media and conditions and represents a novel paradigm approach for the high-throughput process development of recombinant proteins. PMID- 16369982 TI - Inadequacies of the current American Joint Committee on cancer staging system for prostate cancer. AB - BACKGROUND: Two major objectives of the American Joint Committee on Cancer (AJCC) staging system are to ensure appropriate treatments for patients and to determine prognosis. AJCC stage for distant prostate cancer includes patients with regional lymph node involvement. In the current study, the authors assessed whether patients with lymph node involvement and patients with distant metastasis, as determined using the Surveillance, Epidemiology, and End Results (SEER) staging system, had similar treatment and survival duration and, thus, were grouped together appropriately in the AJCC system. METHODS: In total, 4141 patients were selected from The University of Texas M. D. Anderson Cancer Center's Tumor Registry who initially had registered at the center between January 1, 1982, and December 31, 2001, with a diagnosis of prostate cancer; had received no treatment before presentation; and had received treatment at the center. Patients with unknown stage and patients with any other primary malignancies were excluded. Descriptive analyses of demographic and disease variables were performed. Using SEER stage groups, survival analyses and Cox proportional hazards regression analyses were performed. RESULTS: Treatments differed between patients with lymph node involvement and patients with distant metastasis. The median survival was 134 months for patients with lymph node involvement and 42 months for patients with distant metastasis. When these 2 groups were combined, as in the AJCC scheme, the median survival was 86 months. CONCLUSIONS: The treatment and median survival of patients with lymph node involvement differed substantially from those of patients with distant metastasis. The current AJCC scheme for prostate cancer appeared to be inappropriate when considering its purpose, and the authors concluded that it should be revised. PMID- 16369983 TI - Maximal COX-2 immunostaining and clinical response to celecoxib and interferon alpha therapy in metastatic renal cell carcinoma. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) plays a major role in the development of cancer through numerous mechanisms. COX-2 is expressed in the majority of renal cell carcinoma (RCC) tumors and correlates with stage, grade, and microvessel density. Based on potential additive or synergistic antitumor effects, interferon alpha (IFNalpha) and celecoxib, an oral COX-2 inhibitor, were given to metastatic RCC patients in a Phase II trial. METHODS: Patients with untreated, metastatic RCC received IFNalpha 3 million units (MU) daily and celecoxib 400 mg orally (p.o.) twice daily continuously until disease progression or unacceptable toxicity. Pretreatment, paraffin-embedded RCC tumor samples were immunohistochemically stained for COX-2 expression and plasma basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) levels were assayed to determine predictive or prognostic potential. RESULTS: There were three partial responses among 25 patients treated (objective response rate, 12%; 95% confidence interval [CI], 3-31%). The observed median time to disease progression (TTP) for the entire cohort was 3.3 months. A significant association between maximal COX-2 staining and clinical response was observed: all patients who experienced an objective response demonstrated 3+ COX-2 tumor immunostaining (trend test: P=0.03). Therapy was well tolerated without cardiac or other notable toxicity. CONCLUSIONS: The addition of celecoxib to IFNalpha did not increase the objective response rate or TTP of this unselected cohort. Maximal COX-2 tumor immunostaining may identify RCC patents more likely to achieve clinical benefit with COX-2 inhibition in combination with IFNalpha. Further investigation of this combination in 3+ COX-2-overexpressing RCC tumors is warranted. PMID- 16369984 TI - The clinical phenotype of wild-type, heterozygous, and homozygous JAK2V617F in polycythemia vera. AB - BACKGROUND: Several studies have recently reported on the occurrence of a JAK2(V617F) mutation in myeloid cells from the majority of patients with polycythemia vera (PV). The clinical relevance of this novel observation currently is under study. METHODS: In a single institutional study, mutation screening for JAK2(V617F) was performed in DNA derived from archived blood granulocytes from 63 consecutive patients with PV in whom current diagnostic criteria were strictly applied and the diagnosis confirmed by bone marrow histology. RESULTS: The JAK2(V617F) mutant allele was detected in 58 of the 63 patients (92%) with 21% homozygosity. The clinical phenotype of the five patients with the wild-type allele was otherwise typical for the disease. A statistical comparison between JAK2(V617F) heterozygotes (n=45 patients) and homozygotes (n=13 patients) did not reveal any significant associations with regard to age, gender, leukocyte or platelet count at the time of diagnosis, duration of disease, or the incidences of thrombosis or bleeding. However, compared with their heterozygote counterparts, JAK2(V617F) homozygote patients displayed a significantly higher hemoglobin level at the time of diagnosis (P=0.001), an increased incidence of pruritus (69% vs. 38%; P=0.04), a higher rate of fibrotic transformation (23% vs. 2%; P=0.009), and higher PRV-1 transcript levels in their blood granulocytes (P=0.07). CONCLUSIONS: The results of the current clinical study support previous laboratory observations that link JAK2(V617F) with the PV phenotype by demonstrating a mutant allele dose effect on erythrocytosis and clinical and laboratory features characteristic of PV. PMID- 16369985 TI - Surgery by consultant gynecologic oncologists improves survival in patients with ovarian carcinoma. AB - BACKGROUND: Consultant gynecologic oncologists from the regional Comprehensive Cancer Center assisted community gynecologists in the surgical treatment of patients with ovarian carcinoma when they were invited. For this report, the authors evaluated the effects of primary surgery by a gynecologic oncologist on treatment outcome. METHODS: The hospital files from 680 patients with epithelial ovarian carcinoma who were diagnosed between 1994 and 1997 in the northern part of the Netherlands were abstracted. Treatment results were analyzed according to the operating physician's education by using survival curves and univariate and multivariate Cox regression analyses. RESULTS: Primary surgery was performed on 184 patients by gynecologic oncologists, and on 328 patients by general gynecologists. Gynecologic oncologists followed surgical guidelines more strictly compared with general gynecologists (patients with International Federation of Gynecology and Obstetrics [FIGO] Stage I-II disease, 55% vs. 33% [P=0.01]; patients with FIGO Stage III disease, 60% vs. 40% [P=0.003]) and more often removed all macroscopic tumor in patients with FIGO Stage III disease (24% vs. 12%; P=0.02). When patients were stratified according to FIGO stage, the 5-year overall survival rate was 86% versus 70% (P=0.03) for patients with Stage I-II disease and 21% versus 13% (P=0.02) for patients with Stage III-IV disease who underwent surgery by gynecologic oncologists and general gynecologists, respectively. The hazards ratio for patients who underwent surgery by gynecologic oncologists was 0.79 (95% confidence interval [95%CI], 0.61-1.03; adjusted for patient age, disease stage, type of hospital, and chemotherapy); when patients age 75 years and older were excluded, the hazards ratio fell to 0.71 (95% CI, 0.54-0.94) in multivariate analysis. CONCLUSIONS: The surgical treatment of patients with ovarian carcinoma by gynecologic oncologists occurred more often according to surgical guidelines, tumor removal more often was complete, and survival was improved. PMID- 16369986 TI - Extranodal natural killer/T-cell lymphoma, nasal type: the significance of radiotherapeutic parameters. AB - BACKGROUND: The objective of this study was to investigate the correlation between local recurrence and radiotherapeutic parameters, including dose and RT radiotherapy (RT) field. METHODS: The current study included 35 patients who were diagnosed with immunohistochemically confirmed nasal natural killer (NK)/T-cell lymphoma between 1976 and 2004. There were 21 males and 14 females, and they ranged in age from 18 years to 76 years (median, 51 yrs). The primary tumor originated in the nasal cavity in 28 patients, and 32 patients had Stage I disease. Seventeen patients received treatment solely with RT, and the remaining 18 patients received a combination of chemotherapy and RT. The median tumor dose was 50 grays (Gy) (range, 22-60 Gy). Twenty-seven patients received RT to include all macroscopic lesions, all paranasal sinuses, the palate, and the nasopharynx. Eight patients received RT to all macroscopic lesions with generous margins. RESULTS: A complete remission (CR) or a CR/unconfirmed was achieved in 28 patients (80%). The 5-year overall survival (OAS) rate, disease-free survival (DFS) rate, and local control probability (LCP) were 47.3%, 42.9%, and 65.2%, respectively. Patients who received RT only to macroscopic lesions fared less well in terms of LCP (LCP 5 years, 71.9% vs. 41.7%; P=0.007). The difference in RT field also affected both the OAS rate and the DFS rate. Patients who received RT doses>or=50 Gy tended to achieve favorable local control. CONCLUSIONS: In the management of nasal NK/T-cell lymphoma, the RT field affected treatment outcomes. RT doses>or=50 Gy resulted in favorable local control. PMID- 16369987 TI - Prognosis in transplant-eligible patients with agnogenic myeloid metaplasia: a simple CBC-based scoring system. AB - BACKGROUND: Allogeneic hematopoietic stem cell transplantation is potentially curative in agnogenic myeloid metaplasia (AMM) but is associated with substantial mortality and morbidity that necessitates accurate identification of patients in whom benefit outweighs risk. The current study describes the natural history of AMM in transplant-eligible patients and proposes a new prognostic scoring system that favorably compares with other established models. METHODS: Patients diagnosed with AMM before the age of 60 years and seen at Mayo Clinic were identified and the diagnosis confirmed. Relevant demographic, clinical, and laboratory characteristics were abstracted, and the impact of various parameters on overall survival (OS) was evaluated with univariate and multivariate analyses. RESULTS: A cohort of 160 patients with AMM is described. OS was 78 months. Multivariate analysis identified a hemoglobin level of <10 g/dL, white blood cell count of either <4 or >30x10(9)/L, platelet count of <100x10(9)/L, presence of constitutional symptoms, and hepatomegaly as independent predictors of inferior survival. The first 3 complete blood count-based parameters were combined into a new scoring system that resulted in median survivals of 155, 69, and 24 months in the presence of 0, 1, or >or=2 adverse features. The chi-square value for the new model was 80.6 compared with 51.4, 48.4, and 43.7 for the models by Dupriez, Cervantes, and Visani, respectively. CONCLUSIONS: A new scoring system based on blood count at the time of diagnosis can adequately stratify by risk transplant eligible patients with AMM and can accurately identify high-risk as well as intermediate-risk disease. The new system displayed a stronger discriminative value, between risk categories, compared with currently existing prognostic models. PMID- 16369988 TI - Dysplastic nodules frequently develop into hepatocellular carcinoma in patients with chronic viral hepatitis and cirrhosis. AB - BACKGROUND: Advances in imaging technology have enhanced the detection of small nodular lesions during the course of chronic liver disease. METHODS: Between 1995 and 2002, the authors examined 154 consecutive patients with small hepatic nodules without hepatocellular carcinoma (HCC) over a median duration of 2.8 years. The median size of these nodules was 14 mm (range, 7-40 mm). The initial histopathologic diagnosis included high-grade dysplastic nodule (HGDN) (n=13), low-grade dysplastic nodule (LGDN) (n=42), and regenerative nodule (RN) (n=99). RESULTS: A total of 29 (18.8%) nodules developed into HCC during the observation period. Cumulative HCC development rates at the first, third, and fifth year were 46.2%, 61.5%, and 80.8% for HGDN; 2.6%, 30.2%, and 36.6% for LGDN; and 3.3%, 9.7%, and 12.4% for RN, respectively. The rate of HCC development was significantly higher in the HGDN group than for other types (P<0.001). Multivariate analysis disclosed that histopathologic diagnosis (P<0.001) and findings on computed tomographic arterial portography (CT-AP) (P=0.004) were significantly associated with future HCC development. The hazard ratios of HGDN and LGDN were 16.8 (95% confidence interval [CI], 6.19-45.6) and 2.96 (95% CI, 1.20-7.31), respectively. A decrease in portal blood flow also showed a significantly high hazard ratio of 3.04 (95% CI, 1.42-6.50). Approximate annual development rate to HCC was 20% in patients with HGDN and 10% in LGDN. CONCLUSION: HGDN should be considered a precancerous lesion when it appears during follow-up of chronic viral hepatitis or cirrhosis. Reduced portal blood flow in the nodule on computed tomography-AP is also an important predictor for development of hepatocellular carcinoma. PMID- 16369989 TI - Phase II study of a protracted irinotecan schedule in children with refractory or recurrent soft tissue sarcoma. AB - BACKGROUND: Irinotecan (CPT-11) is a novel antineoplastic agent that takes effect by inhibiting topoisomerase I. The Italian Soft Tissue Sarcoma (STS) Committee performed a multiinstitutional Phase II study to evaluate its effect on STS. METHODS: Over a 2-year period between 2002 and 2004, 32 heavily pretreated patients were administered 60-minute infusions of irinotecan at 20 mg/m2/day, for 5 days a week, for 2 consecutive weeks. The courses were repeated every 4 weeks for at least 2 courses, unless there were signs of toxicity or disease progression. Thirty patients, 13 with peripheral primitive neuroectodermal tumor (PNET), 12 with rhabdomyosarcoma (RMS), 3 with desmoplastic small round cell tumor (DSRCT), and 2 with other STS were evaluable for response. RESULTS: A total of 79 cycles were delivered. The main regimen-related toxicity was diarrhea, occurring in 58% of cycles with 9 episodes graded as 3 or 4. Grade 3-4 neutropenia was recorded in 10% of cycles. The overall response rate was 23% (2 complete remissions +5 partial remissions of 30 patients), 38% for PNET and 16% for RMS. In addition, 4 minor responses were noted. CONCLUSIONS: As a single agent in the treatment of recurrent and refractory STS, irinotecan administered on a daily x5 x2 schedule revealed a noteworthy response rate in a population of heavily pretreated patients, especially in the subset of patients with PNET. Its hematologic toxicity profile warrants further investigation in association with other myelotoxic agents. PMID- 16369990 TI - Sociodemographic determinants of cancer treatment health literacy. PMID- 16369992 TI - Telomerase-specific T-cells kill pancreatic tumor cells in vitro and in vivo. AB - BACKGROUND: Adoptive cell transfer is described as an innovative and challenging option for the treatment of malignant melanoma. In the current study, the generation and expansion of telomerase-specific T-cells for adoptive cell transfer and their use in a syngeneic pancreatic carcinoma mouse model was investigated. METHODS: Telomerase-specific T-cells were generated either in vitro by coculture of human lymphocytes with telomerase-peptide-pulsed dendritic cells or in vivo by injection of peptide plus adjuvant into C57BL/6 mice. Spleens were harvested after immunization and lymphocytes were expanded in the presence of feeder cells. T-cells were tested in vitro against human leukocyte antigen (HLA) matched, telomerase-positive pancreatic carcinoma cells. Tumor-bearing (subcutaneous) mice pretreated with cyclophosphamide were injected intravenously with the expanded cells. RESULTS: It was possible to generate and expand telomerase-specific T-cells with cytotoxic activity. The protocol did not work as well in the murine setting. However, adoptive cell transfer with murine antigen specific T-cells delayed disease progression in tumor-bearing mice significantly. CONCLUSIONS: Generation of antigen-specific T-cells is feasible; the expansion of these cells could be accomplished without loss of function. Antigen-specific T cells demonstrated significant cytotoxic activity in a syngeneic, subcutaneous mouse model. However, further optimization of the expansion protocol is warranted. PMID- 16369993 TI - Societies, journals, and editors. PMID- 16369994 TI - Ocular toxicity during adjuvant chemoendocrine therapy for early breast cancer: results from International Breast Cancer Study Group trials. AB - BACKGROUND: Others have reported ocular toxicity after adjuvant chemoendocrine therapy, but this study looked at ocular toxicity in similarly treated patients from large randomized clinical trials. METHODS: Information was retrieved on incidence and timing of ocular toxicity from the International Breast Cancer Study Group (IBCSG) database of 4948 eligible patients randomized to receive tamoxifen or toremifene alone or in combination with chemotherapy (either concurrently or sequentially). Case reports of patients with ocular toxicity were evaluated to determine whether ocular toxicity occurred during chemotherapy and/or hormonal therapy. Additional information was obtained from participating institutions for patients in whom ocular toxicity occurred after chemotherapy but during administration of tamoxifen or toremifene. RESULTS: Ocular toxicity was reported in 538 of 4948 (10.9%) patients during adjuvant treatment, mainly during chemotherapy. Forty-five of 4948 (0.9%) patients had ocular toxicity during hormone therapy alone, but only 30 (0.6%) patients had ocular toxicity reported either without receiving any chemotherapy or beyond 3 months after completing chemotherapy and, thus, possibly related to tamoxifen or toremifene. In 3 cases, retinal alterations, without typical aspects of tamoxifen toxicity, were reported; 4 patients had cataract (2 bilateral), 12 impaired visual acuity, 10 ocular irritation, 1 optical neuritis, and the rest had other symptoms. CONCLUSION: Ocular toxicity during adjuvant therapy is a common side effect mainly represented by irritative symptoms due to chemotherapy. By contrast, ocular toxicity during hormonal therapy is rare and does not appear to justify a regular program of ocular examination. However, patients should be informed of this rare side effect so that they may seek prompt ophthalmic evaluation for ocular complaints. PMID- 16369995 TI - Clinically significant prognostic factors for differentiated thyroid carcinoma: a population-based, nested case-control study. AB - BACKGROUND: Different scoring systems currently are being used to stratify patients with differentiated thyroid carcinoma (DTC) into risk groups. DTC is usually subdivided into papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC). The objective of the current study was to identify those factors that predict long-term unfavorable prognosis and to evaluate the predictive accuracy of the TNM staging system. METHODS: The authors conducted a nested case control study within the cohort of all patients (n=5123) diagnosed with DTC in Sweden between 1958-1987 who survived at least 1 year after diagnosis. One control, matched by age at diagnosis, gender, and calendar period, was randomly selected for each case (patients who died of DTC). All patients were classified at the time of diagnosis according to the TNM staging system. The effect of prognostic factors on DTC mortality was evaluated using conditional logistic regression. RESULTS: Patients with widely invasive FTC experienced a significantly higher mortality compared with PTC patients. The grade of differentiation was found to influence mortality significantly. Patients with TNM Stage IV disease had a higher mortality rate compared with patients with Stage II disease (odds ratio [OR]=9.1; 95% confidence interval [95% CI], 5.7-14.6). Patients with lymph node metastases experienced a higher mortality (OR=2.5; 95% CI, 1.6-4.1) and patients with distant metastasis at the time of diagnosis were found to have a nearly 7-fold higher mortality rate (OR=6.6; 95% CI, 4.1-10.5). Incomplete surgical excision was associated with higher mortality, particularly in patients with Stage I disease. CONCLUSIONS: In the current study, the following were found to be clinically significant prognostic factors for patients with DTC: histopathologic subgroup, TNM staging including lymph node metastases and distant metastases, and completeness of the surgical excision. PMID- 16369996 TI - Phenotypic changes associated with DYNACTIN-2 (DCTN2) over expression characterise SJSA-1 osteosarcoma cells. AB - DYNACTIN-2 (DCTN2) localises to chromosome 12q13-q15, a region prone to stable amplification in several cancers. Transient DCTN2 overexpression has a significant impact on cellular phenotype primarily due to disruption of the DYNEIN-dynactin motor. Changes reported include alterations of microtubule directed movement of molecular (e.g. TP53) and organelle (e.g. Golgi) cargoes towards the nucleus, centrosome biology, cellular movement and mitosis with a potential predisposition to mitotic block and polyploidy. These changes would be expected to be of relevance to carcinogenesis. To investigate this, we report the first study of DCTN2 genomic amplification and sustained DCTN2 overexpression in cancer cells. QFMPCR was employed to characterise the extent of chromosome 12q13 q15 amplicons in SJSA-1, SJRH30, U373MG and CCF-STTG1 cancer cells. DCTN2 amplification was present in SJSA-1, U373MG and SJRH30 cells, yet was incomplete at the 5'-end in SJRH30 cells. Only SJSA-1 cells were characterised by DCTN2 overexpression on Western blot analyses. Microscopy studies distinguished SJSA-1 cells by greater DCTN2 immunofluorescence and diminished centrosome and 58K protein Golgi-marker focus compared to SJRH30 cells. Indirect evidence derived from the published work of others indicated that TP53 transport into the nucleus was unimpaired. Furthermore, we observed that SJSA-1 cells were easy to propagate. In conclusion, persistent DCTN2 overexpression can be tolerated in SJSA-1 cancer cells despite phenotypic abnormalities predicted from transient overexpression studies. This preliminary study does not support a major role for DCTN2 overexpression in carcinogenesis, although further studies would be necessary to confirm this. PMID- 16369997 TI - Apple flavonoids inhibit growth of HT29 human colon cancer cells and modulate expression of genes involved in the biotransformation of xenobiotics. AB - Flavonoids from fruits and vegetables probably reduce risks of diseases associated with oxidative stress, including cancer. Apples contain significant amounts of flavonoids with antioxidative potential. The objectives of this study were to investigate such compounds for properties associated with reduction of cancer risks. We report herein that apple flavonoids from an apple extract (AE) inhibit colon cancer cell growth and significantly modulate expression of genes related to xenobiotic metabolism. HT29 cells were treated with AE at concentrations delivering 5-50 microM of one of the major ingredients, phloridzin ("phloridzin-equivalents," Ph.E), to the cell culture medium, with a synthetic flavonoid mixture mimicking the composition of the AE or with 5-100 microM individual flavonoids. HT29 cell growth was inhibited by the complex extract and by the mixture. HT29 cells were treated with nontoxic doses of the AE (30 microM, Ph.E) and after 24 h total RNA was isolated to elucidate patterns of gene expression using a human cDNA-microarray (SuperArray) spotted with 96 genes of drug metabolism. Treatment with AE resulted in an upregulation of several genes (GSTP1, GSSTT2, MGST2, CYCP4F3, CHST5, CHST6, and CHST7) and downregulation of EPHX1, in comparison to the medium controls. The enhanced transcriptional activity of GSTP1 and GSTT2 genes was confirmed with real-time qRT-PCR. On the basis of the pattern of differential gene expression found here, we conclude that apple flavonoids modulate toxicological defense against colon cancer risk factors. In addition to the inhibition of tumor cell proliferation, this could be a mechanism of cancer risk reduction. PMID- 16369998 TI - Intestinal absorption and presystemic disposition of sildenafil citrate in the rabbit: evidence for site-dependent absorptive clearance. AB - Sildenafil citrate is the first oral treatment for erectile dysfunction. Its oral bioavailability is about 40%. This research investigated the intestinal transport parameters of sildenafil citrate in rabbit using an in situ intestinal perfusion technique. This was studied in four different anatomical sites, namely duodenum, jejunoileum, ascending colon and rectum. The results revealed the highest absorptive clearance in the jejunoileum. The values of the permeability area product normalized to segment length (ml/min.cm) were 0.0101, 0.0063, 0.0059 and 0.0023 and those of the percentage absorbed were 68.0, 32.3, 23.0 and 5.0 in jejunoileum, duodenum, ascending colon and rectum, respectively. The values of the length (cm) required for complete absorption were 87.6, 137, 153 and 384 for each anatomical site in the same order. The absorptive clearance did not correlate with the net water flux in the four anatomical regions studied, indicating a mainly passive diffusion mechanism through a transcellular pathway. The plasma sildenafil concentrations achieved during intestinal perfusion experiments and sildenafil total body clearance in the rabbit were used to calculate the fraction of sildenafil that reached the systemic circulation relative to the amount that disappeared from the intestinal segment. Only 34% of sildenafil that disappeared from the intestinal segment appeared in the systemic circulation indicating that the presystemic elimination of sildenafil is 66%. These results confirm that the incomplete bioavailability of sildenafil is mainly due presystemic elimination. PMID- 16370000 TI - Accuracy of computed tomography in the detection of blunt bowel and mesenteric injuries (Br J Surg 2005; 92: 1024-1028). PMID- 16370001 TI - Letter 1: Randomized clinical trial of vertical or transverse laparotomy for abdominal aortic aneurysm repair (Br J Surg 2005; 92: 1208-1211). PMID- 16370002 TI - Letter 2: Randomized clinical trial of vertical or transverse laparotomy for abdominal aortic aneurysm repair (Br J Surg 2005; 92: 1208-1211). PMID- 16370003 TI - A close look at short C-CH3...potassium contacts: synthetic and theoretical investigations of [M2Co2(mu3-OtBu)2(mu2-OtBu)4(thf)n] (M = Na, K, Rb, thf = tetrahydrofuran). AB - Agostic interactions of the type Si-CH3M+ (M = alkali metal) are frequently mentioned in discussions of solid-state structures of trimethylsilyl compounds and the purpose of this work was to elucidate if they also exist in the related tert-butyl species by using density functional theory. The compounds [M2Co2(mu3 OtBu)2(mu2-OtBu)4(thf)n] (M = Na, n = 2; M = K, n = 0; M = Rb, n = 1) have been synthesised and their crystal structures determined. Close contacts of methyl groups with K atoms are observed in the solid-state structure of [K2Co2(mu3 OtBu)2(mu2-OtBu)4], and calculations of the rotational barrier of a tert-butoxy group about the axis through the C-O bond were performed. It was shown that apparent short C-CH3K distances are in this case a consequence of the packing in the extended solid-state structure. PMID- 16370004 TI - Pyrazole complexes as anion receptors. AB - The behavior of the receptors [Re(CO)3(Hdmpz)3]BAr'4 (Hdmpz = 3,5 dimethylpyrazole) (1) and [Re(CO)3(HtBupz)3]BAr'4 (HtBupz = 3(5)-tert butylpyrazole) (2; Ar' = 3,5-bis(trifluoromethyl)phenyl) toward the anions fluoride, chloride, bromide, iodide, hydrogensulfate, dihydrogenphosphate, nitrate, and perrhenate was studied in CD3CN solution. In most cases, the receptors were stable. Anion exchange was fast, and binding constants were calculated from the NMR titration profiles. The structure of the adduct [Re(CO)3(HtBupz)3] x NO3 (3) was determined by X-ray diffraction. Two pyrazole moieties are hydrogen-bonded to one nitrate oxygen atom, and the third pyrazole moiety is hydrogen-bonded to an oxygen atom of an adjacent nitrate, leading to infinite chains. The structure of the adduct [Re(CO)3(Hdmpz)3]BAr'4acetone (4), also determined by X-ray diffraction, showed a similar interaction of two pyrazole N-H groups with the acetone oxygen atom. F- and H2PO4(-) deprotonate the receptors, and HSO4(-) decomposed 1. The structure of one of the decomposition products (5), determined by X-ray diffraction, is consistent with pyrazole protonation and substitution by sulfate. PMID- 16370006 TI - Versatile coordination chemistry towards multifunctional carbon nanotube nanohybrids. AB - Dispersible single-walled carbon nanotubes grafted with poly(4-vinylpyridine), SWNT-PVP, were tested in coordination assays with zinc tetraphenylporphyrin (ZnP). Kinetic and spectroscopic evidence corroborates the successful formation of a SWNT-PVPZnP nanohybrid. Within this SWNT-PVPZnP nanohybrid, static electron transfer quenching (2.0+/-0.1) x 10(9) s(-1) converts the photoexcited-ZnP chromophore into a radical-ion-pair state with a microsecond lifetime, namely one electron oxidized-ZnP and reduced-SWNT. PMID- 16370005 TI - Fine tuning of the oxidation locus, and electron transfer, in nickel complexes of pro-radical ligands. AB - A large number of complexes of the first-row transition metals with non-innocent ligands has been characterized in the last few years. The localization of the oxidation site in such complexes can lead to discrepancies when electrons can be removed either from the metal center (leading to an M((n+1)+) closed-shell ligand) or from the ligand (leading to an M(n+) open-shell ligand). The influence of the ligand field on the oxidation site in square-planar nickel complexes of redox-active ligands is explored herein. The tetradentate ligands employed herein incorporate two di-tert-butylphenolate (pro-phenoxyl) moieties and one orthophenylenediamine spacer. The links between the spacer and both phenolates are either two imines ([Ni(L1)]), two amidates ([Ni(L3)]2-), or one amidate and one imine ([Ni(L2)]-). The structure of each nickel(II) complex is presented. In the noncoordinating solvent CH2Cl2, the one-electron-oxidized forms are ligand radical species with a contribution from a singly occupied d orbital of the nickel. In the presence of an exogenous ligand, such as pyridine, a Ni(III) closed-shell ligand form is favored: axial ligation, which stabilizes the trivalent nickel in its octahedral geometry, induces an electron transfer from the metal(II) center to the radical ligand. The affinity of pyridine for the phenoxylnickel(II) species is correlated to the N-donor ability of the linkers. PMID- 16370007 TI - Unexpected structural diversity in alkali metal azide-crown ether complexes: syntheses, X-ray structures, and quantum-chemical calculations. AB - A series of alkali metal azide-crown ether complexes, [Li([12]crown-4)(N3)], [Na([15]crown-5)(N3)], [Na([15]crown-5)(H2O)2]N3, [K([18]crown-6)(N3)(H2O)], [Rb([18]crown-6)(N3)(H2O)], [Cs([18]crown-6)(N3)]2, and [Cs([18]crown 6)(N3)(H2O)(MeOH)], has been synthesised. In most cases, single crystals were obtained, which allowed X-ray crystal structures to be derived. The structures obtained have been compared with molecular structures computed by density functional theory (DFT) calculations. This has allowed the effects of the crystal lattice on the structures to be investigated. Also, a study of the M-N(terminal) metal-azide bond length and charge densities on the metal (M) and terminal nitrogen centre (N(terminal)) in these complexes has allowed the nature of the metal-azide bond to be probed in each case. The bonding in these complexes is believed to be predominantly ionic or ion-dipole in character, with the differences in geometries reflecting the balance between maximising the coordination number of the metal centre and minimising ligand-ligand repulsions. The structures of the crown ether complexes determined in this work show the subtle interplay of such factors. The significant role of hydrogen bonding is also demonstrated, most clearly in the structures of the K and Rb dimers, but also in the chain structure of the hydrated Cs complex. PMID- 16370008 TI - Stabilization of reactive organometallic intermediates inside a self-assembled nanoscale host. PMID- 16370009 TI - Crystal-to-crystal phase transition in self-assembled mesoporous iron oxide films. PMID- 16370010 TI - Programmable cell adhesion encoded by DNA hybridization. PMID- 16370011 TI - Enantioselective alpha-fluorination of carbonyl compounds: organocatalysis or metal catalysis? PMID- 16370012 TI - An eta3-H2SiR2 Adduct of [[PhB(CH2PiPr2)3]Fe(II)H]. PMID- 16370013 TI - Stacked-cup carbon nanotubes for photoelectrochemical solar cells. PMID- 16370014 TI - Compounds with the "maple leaf" lattice: synthesis, structure, and magnetism of M(x)[Fe(O2CCH2)2NCH2PO3]6n.H2O. PMID- 16370015 TI - Heterogeneous catalysis with nickel-on-graphite (Ni/C(g)): reduction of aryl tosylates and mesylates. PMID- 16370016 TI - Direct carbon-carbon bond formation from alcohols and active methylenes, alkoxyketones, or indoles catalyzed by indium trichloride. PMID- 16370017 TI - The fluorinase from Streptomyces cattleya is also a chlorinase. PMID- 16370018 TI - Carbon nanotube delivery of the GFP gene into mammalian cells. PMID- 16370019 TI - Forster resonance energy transfer investigations using quantum-dot fluorophores. AB - Forster resonance energy transfer (FRET), which involves the nonradiative transfer of excitation energy from an excited donor fluorophore to a proximal ground-state acceptor fluorophore, is a well-characterized photophysical tool. It is very sensitive to nanometer-scale changes in donor-acceptor separation distance and their relative dipole orientations. It has found a wide range of applications in analytical chemistry, protein conformation studies, and biological assays. Luminescent semiconductor nanocrystals (quantum dots, QDs) are inorganic fluorophores with unique optical and spectroscopic properties that could enhance FRET as an analytical tool, due to broad excitation spectra and tunable narrow and symmetric photoemission. Recently, there have been several FRET investigations using luminescent QDs that focused on addressing basic fundamental questions, as well as developing targeted applications with potential use in biology, including sensor design and protein conformation studies. Herein, we provide a critical review of those developments. We discuss some of the basic aspects of FRET applied to QDs as both donors and acceptors, and highlight some of the advantages offered (and limitations encountered) by QDs as energy donors and acceptors compared to conventional dyes. We also review the recent developments made in using QD bioreceptor conjugates to design FRET-based assays. PMID- 16370020 TI - Sensitized emission of luminescent lanthanide complexes based on 4-naphthalen-1 yl-benzoic acid derivatives by a charge-transfer process. AB - The photophysical properties of 4-naphthalen-1-yl-benzoic acid ligands and their Eu(III)-cored complexes were systematically investigated to elucidate the effective energy-transfer pathway in luminescent lanthanide complexes. A series of 4-naphthalen-1-yl-benzoic acid ligands, such as 4-naphthalen-1-yl-benzoic acid (NA-1), 4-[4-(4-methoxyphenyl)-naphthalen-1-yl]-benzoic acid (NA-2), and 4-{4-[4 (4-methoxyphenyl)-naphthalen-1-yl]-benzyloxy}-benzoic acid (NA-3), were synthesized and utilized for the synthesis of their Eu(III)-cored complexes, corresponding to NAC-1, NAC-2, and NAC-3. The fluorescence spectra of NA-1 and NA 2 show large Stokes shifts with increasing solvent polarity. These large Stokes shifts might be dominantly due to the formation of an intramolecular charge transfer (ICT) complex in the excited state. Also, the intensive luminescence of the Eu(III) ions by the photoexcitation of the ligand in NAC-1 and NAC-2 in polar solvents supports that the energy transfer from the ligand to the Eu(III) ion takes place efficiently. In the case of NA-3, which has a -CH2OPh- group that acts as a blocking group, there is no dependence of the fluorescence spectrum on the solvent nature and no luminescence of the Eu(III) ions by the photoexcitation of the ligand, indicating no formation of the ICT state. This can be due to the fact that the formation of the ICT state in NA-3 was prevented because the -OCH2- group acts as a blocking group by interrupting the pi-conjugation between the benzoic acid and the naphthalene unit. From these photophysical studies, we suggest that the ICT state plays a very important role in the energy-transfer pathway from the ligand to the Eu(III) ion. To our best knowledge, this is the first demonstration of sensitized emission of luminescent lanthanide complexes based on 4-naphthalen-1-yl-benzoic acid derivatives by the charge-transfer process. PMID- 16370022 TI - Prognostic significance of alarm symptoms in patients with gastric cancer (Br J Surg 2005; 92: 840-846). PMID- 16370023 TI - Randomized clinical trial comparing 5-year recurrence rate after laparoscopic versus Shouldice repair of primary inguinal hernia (Br J Surg 2005; 92: 1085 1091). PMID- 16370025 TI - The protective effects of caffeic acid phenethyl ester (CAPE) against liver damage induced by cigarette smoke inhalation in rats. AB - The aim of this study was to investigate the histological and biochemical changes in liver of rats exposed to cigarette smoke and effects of caffeic acid phenetyl ester (CAPE) on these changes. For this purpose, 21 male Wistar rats were divided into three groups. Animals in Group I were used as control. Rats in Group II were exposed to cigarette smoke and rats in Group III were exposed to cigarette smoke and injected daily with CAPE. At the end of the 60-days experimental period, all rats were killed by decapitation and blood samples were obtained. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin levels and hepatic superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px ), malondialdehyde (MDA) contents were determined. Following routine histological procedures, liver tissue specimens were examined under a light microscope. The levels of ALT, AST, total bilirubin, SOD, GSH-Px and MDA were significantly increased in rats exposed to cigarette smoke compared with those of the controls. Light microscopic examination of liver specimens from rats exposed to cigarette smoke revealed mononuclear cell infiltration and that some of the hepatocytes had a hyperchromatic nucleus and enlarged sinusoids. The rats which were treated with CAPE along with cigarettes had partially attenuated histological changes associated with cigarette exposure. In conclusion, the damage inflicted by cigarette in the rat liver can be partially prevented by CAPE administration. PMID- 16370026 TI - Canadian developments. New medical marijuana regulations approved. PMID- 16370027 TI - Canadian developments. Minister announces new drug approval initiatives. PMID- 16370028 TI - UK: Guidelines for liver transplants for HIV-positive people issued. PMID- 16370029 TI - UK: House Committee urges universal free HIV treatment. PMID- 16370030 TI - China: alleged quarantine of HIV-positive people in several provinces. PMID- 16370031 TI - Kenya: Free antiretrovirals distributed in prison. PMID- 16370032 TI - South Africa: Prison oversight body recommends permitting consensual sex between inmates. PMID- 16370033 TI - Scotland: Executive proposes mandatory HIV testing for criminal suspects. PMID- 16370034 TI - HIV/AIDS in the courts--Canada. Criminal law and HIV transmission or exposure: one new case. HIV-positive man receives three years in prison for unprotected sex. PMID- 16370035 TI - HIV/AIDS in the courts--Canada. Red Cross pleads guilty to distributing tainted blood. PMID- 16370036 TI - China: public health officials convicted of malpractice. PMID- 16370037 TI - Closed ambulatory lateral internal sphincterotomy for chronic anal fissures. PMID- 16370038 TI - Giant rectal diverticulum presenting as an ischioanal abscess. PMID- 16370039 TI - Parasacral and perineal sphincteroplasty. PMID- 16370040 TI - Managing patients undergoing non-cardiac surgery: need to shift emphasis from risk stratification to risk modification. AB - Many patients undergo non-invasive testing for the detection of coronary artery disease before non-cardiac surgery. This is despite the low predictive value of positive tests in this population and the lack of any evidence of benefit of coronary revascularisation before non-cardiac surgical procedures. Further, this strategy often triggers a clinical cascade exposing the patient to progressively riskier testing and intervention and results in increased costs and unnecessary delays. On the other hand, administration of beta blockers, and more recently statins, has been shown to reduce the occurrence of perioperative ischaemic events. Therefore, there is a need for a shift in emphasis from risk stratification by non-invasive testing to risk modification by the application of interventions, which prevent perioperative ischaemia--principally, perioperative beta adrenergic blockade and perhaps treatment with statins. Clinical risk stratification tools reliably identify patients at high risk of perioperative ischaemic events and can guide in the appropriate use of perioperative medical treatment. PMID- 16370041 TI - Mental disorders after laryngectomy. PMID- 16370042 TI - Management of colorectal cancer in pregnancy: a multimodality approach. AB - Colorectal cancer (CRC) is one of the 3 most common types of cancer in women, but CRC during pregnancy is rare, with a reported incidence of approximately 0.002%. Synchronous colon cancer during pregnancy presents a diagnostic and therapeutic challenge for clinicians because there are no generally accepted guidelines regarding diagnosis or treatment. The diagnosis is challenging because the presenting signs/symptoms of CRC are often attributed to the usual complications of pregnancy, which could delay the diagnosis and allow the cancer to progress to an advanced stage. Carcinogenesis of colon cancer in pregnancy is not clear, but a few studies suggest that the increased levels of estrogen and progesterone related to pregnancy stimulate the growth of CRC with their receptors. The aim of treatment is to start therapy for the mother as early as possible and to simultaneously deliver the baby at the earliest time allowable. The management mandates a multidisciplinary approach involving experts in obstetrics, neonatology, gastrointestinal surgery, and medical oncology. The medical community should be able to diagnose colon cancer earlier in pregnancy in order to improve prognosis. The primary care physician or obstetrician should refer the pregnant patient with significant gastrointestinal symptoms to the gastroenterologist for evaluation. Likewise, the gastroenterologist should be prepared to perform sigmoidoscopy (preferably without endoscopic medications) for significant lower gastrointestinal symptoms such as persistent rectal bleeding. Herein, the author reviews the literature concerning the diagnosis and treatment of CRC in pregnancy and discusses the role of newer agents approved for the treatment of CRC. PMID- 16370043 TI - The role of EGFR-TK inhibition in non-small cell lung cancer. PMID- 16370044 TI - Delayed enhancement of pelvic lymphangiomyoma associated with lymphangioleiomyomatosis on MR imaging (2005: 9b). PMID- 16370045 TI - Vesicle of soft palate. Superficial mucocele. PMID- 16370046 TI - Evolving health care issues--a primer on fourteen key issues facing providers. PMID- 16370047 TI - [On Q-wave and non-Q wave myocardial infarcts]. AB - Much has been said, and is still being said, on Q-wave and non-Q wave myocardial infarcts, trying to relate this electrocardiographic behavior with the culprit coronary arteries and the location of the damaged myocardium. However, it seems logic to bear in mind that the presence or absence of abnormal Q waves depends on the relation established between the zone of damaged myocardium and the width of the electrical endocardium. It must be recalled that the presence of normal Q waves is possible in leads that seem to move away from the first vector of ventricular activation. Besides, the electrical endocardium, i.e., the territory of distribution of Purkinje's network, is situated mainly in the lower half of the ventricles and is virtually absent in basal regions. This endocardium constitutes a histological-functional entity, since the Purkinje fibers, which receive at the same time the activation impulses, are depolarized simultaneously without producing differences in potential. Therefore, these fibers cannot supply an electrical contribution either in normal condition or in the presence of limited damage. Nevertheless, when the damaged zone reaches beyond the exterior limits of this endocardium, for example, in regions where it is small, the exploring electrode can register abnormal Q waves, due to the activation fronts that are moving away, followed by R waves originated in contiguous bands of non damaged myocardium. We present two characteristic examples of the electrocardiographic manifestations of a transmural left ventricle infarct (QS complexes) and of a subendocardial infarct, reaching beyond the borders of the electrical endocardium (QR complexes). In both of these cases, the electrocardiographic data agree with the anatomical findings. PMID- 16370048 TI - [Primary acute myocarditis. A 10- years institutional experience]. AB - Acute myocarditis (AM) is associated with viral infections: Coxsackie and ECHOviruses among others. Autoimmunity has been proposed as a pathogenic mechanism. Benefit of classic immunosuppression (prednisone-azathioprine) or immunomodulation (monomeric-human IgG) is still uncertain. OBJECTIVE: To review incidence and clinical approach to AM at a Cardiology referral center. MATERIAL AND METHODS: A 10-yeard period (1992-2003) is reviewed. A standard questionary was applied to 49 consecutive patients referred by clinicians with a diagnosis of AM. RESULTS: AM was found in 17 women and 32 men, median age 24 and 28 years, respectively. They presented heart failure with dyspnea/ortopnea (70-47%), peripheral edema/jugular vein plethora (41-37%), chest pain, and tachycardia (50%), NYHA functional class was Ill-IV in 22. The EKG showed sinus tachycardia or conduction defects. Transthoracic echocardiograms in 47 cases showed EF (mean) of 41% with enlarged left ventricle diameter. Antivirus antibodies were present in 54% of those cases studied, Coxsackie or ECHOvirus were identified through a serologic assay. Twenty-nine (61%) of our cases developed dilated cardiomyopathy, three patients died. It is not possible to reach a conclusion regard to immunomodulation therapy, because it was applied to only 12 patients. CONCLUSIONS: At the Instituto Nacional de Cardiologia "I. Chavez", AM depicts an incidence of 1/1,000 patients a year. It is necessary to standardize the clinical approach for diagnosis and treatment, progression to dilated cardiomyopathy and death during acute stage occurs in two-thirds of our patients. PMID- 16370049 TI - [Postero-lateral biventricular infarcts. An electro-anatomical comparison]. PMID- 16370050 TI - How has the sexual behaviour of gay men changed since the onset of AIDS: 1986 2003. AB - OBJECTIVE: To report changes in sexual behaviour among gay men in Sydney from 1986 to 2003. METHODS: Baseline data from four studies of gay men in Sydney were used: the Social Aspects of the Prevention of AIDS study (1986/87: 91 HIV positive and 444 HIV-negative men); the Sydney Men and Sexual Health cohort (1993 95: 237 HIV-positive and 910 HIV-negative men); the Health in Men cohort of HIV negative gay men (2001-03: 1,148 men); the Positive Health cohort of HIV-positive gay men (2001/02: 237 men). Each sample was recruited and interviewed using similar methods. RESULTS: Fewer HIV-positive men had sex with casual partners over time (76.9% in 1986/87 to 63.7% in 2001/02; p = 0.001), but more HIV negative men had sex with 10 or more casual partners in the previous six months (27.7% in 1986/87 to 37.7% in 2001-03; p = 0.012). The proportions engaging in particular sex practices with casual partners changed over time: anal intercourse without condoms that included ejaculation in the rectum fell from 29.4% among HIV positive men and 32.4% among other men in 1986/87 to 17.8% and 10.0% respectively in 1993-95 (p = 0.034 and p < 0.001 respectively), but increased to 37.7% and 18.4% respectively in 2001-03 (p-values < 0.001); rimming one's partner increased from 36.8% among HIV-positive men and 17.6% among other men in 1986/87 to 63.6% and 52.3% respectively in 2001-03 (p = 0.001 and p < 0.001 respectively). CONCLUSION: Gay men's sexual behaviour with casual partners has changed over time, perhaps partly in response to HIV and partly as a general expansion of sexual repertoires. These changes have implications for gay men's health. PMID- 16370051 TI - Prevalence and risk factors of hepatitis C in HIV-negative homosexual men in Sydney, Australia. AB - OBJECTIVE: To determine the prevalence and risk factors for hepatitis C (HCV) in HIV-negative homosexual men in Sydney. METHODS: A cohort study was conducted in a sample of community-based, HIV-negative, homosexual men in Sydney. Participants underwent a face-to-face interview regarding sexual behaviour, sexually transmissible infections, and injecting drug use (IDU). RESULTS: Eight hundred and twenty-four men consented to HCV testing, and the prevalence was 0.85% (95% CI 0.34-1.74). HCV seropositivity was strongly associated with a history of IDU (OR = 60.43, 95% CI 6.70-544.79). All HCV seropositive individuals reported a history of either IDU or other means by which they may have had parenteral exposure to HCV. There was no evidence of an independent association between sexual behaviour and HCV infection. CONCLUSION: The prevalence of HCV in this cohort was about the same as in the general population in Australia, and there was no evidence for sexual transmission in this population. PMID- 16370052 TI - Prodromal angina reduces infarcted mass less in interventionally reperfused than in thrombolysed myocardial infarction. AB - There is a lot of evidence that angina during the 24-48 h before a reperfused myocardial infarction improves the evolution of the patients. However, there are studies that failed to demonstrate this protective effect of preinfarction angina in an interventional reperfusion setting. OBJECTIVE: To compare the effect of preinfarction angina (PIA) on inhospital evolution of thrombolysis vs. interventionally reperfused acute myocardial infarction (AMI). MATERIAL AND METHOD: There were prospectively studied 133 consecutive AMI patients, eligible for reperfusion (thrombolysis or interventional). History of PIA under 48 hours was obtained. Evolution of AMI was evaluated considering the following end points: the ratio between the number of ECG leads with final pathologic Q wave and the number of leads with initial ST elevation, CK-MB values, separate and composite incidence of death, heart failure, shock and incidence of serious arrhythmia (sustained VT or ventricular fibrillation). RESULTS: ECG ratio was lower in patients with PIA (0.511 +/- 0.281 vs. 0.646 +/- 0.274, p=0.02) in thrombolysed patients, but it was higher in interventionally reperfused patients (0.740 +/- 0.418 vs. 0.554 +/- 0.295 p=0.11). CK-MB values were lowered by PIA in thrombolysed AMI (122 +/- 74 vs. 190 +/- 89, p=0.0003), but they were not in the interventional group. Clinical end-points were slightly less frequent in patients with PIA, in both reperfusion groups, but not statistically significant. Major arrhythmia occurred less frequently in interventionally reperfused patients with PIA (9.5% vs. 31.6%, p=0.12). CONCLUSION: Preinfarction angina under 48 hours significantly reduces infarcted mass (measured by ECG and enzymes) in thrombolysed patients, but not in the interventional group. However, PIA reduced arrhythmic end-point in interventional setting. PMID- 16370053 TI - Patients with primary antiphospholipid syndrome and coronary microvascular dysfunction--a distinct clinical subset. AB - Morbidity of patients with cardiac syndrome X (CSX) is high. Impairment of microvascular endothelial function has been suggested to be a mechanism of the disease. The study was undertaken to assess some of the characteristics of patients with primary antiphospholipid syndrome (pAPS) and CSX. METHODS: We studied 36 patients with CSX, 14 patients having pAPS and 10 healthy controls. Patients evaluation included: clinical examination, 12-lead ECG, effort treadmill test (protocol Bruce modified), determination of plasma triglycerides, cholesterol, antiphospholipid antibodies (APLA). There were determined as markers of the inflammatory state: serum phospholipase (PL-A2) and peripheral neutrophils activity. RESULTS: Patients with pAPS presented normal values of serum cholesterol and triglycerides levels, normal PL-A2 activity, moderate superoxide anion generation. Patients without APLA presented hyperlipidemia, increased PL-A2 activity, increased superoxide anion generation. During the follow-up period we found a correlation between P1-A2 activity and ischemic episodes, but only in patients with CSX and pAPS there were registered cardiovascular events. CONCLUSION: Patients with SCX and pAPS represent a distinct clinical subset, being characterized by minimal inflammation, absence of usual risk factors for coronary heart disease, more severe prognosis related to recurrent thromboses and the need for early anticoagulant therapy. PMID- 16370054 TI - [Severe intoxication with verapamil]. AB - Calcium channel blockers are currently widely used to treat many cardiological alterations; however, overdose and poisoning have been associated with morbidity and mortality mainly in those patients with suicidal attempts. We report a case and review the pathophysiology of overdose, treatment, and prognosis. PMID- 16370055 TI - Understanding immigrant Chinese Americans' participation in cancer screening and clinical trials. AB - The purpose of this study was to identify potential barriers and facilitators to Chinese immigrant participation in cancer screening and clinical trials. A series of focus groups, in English, Cantonese, and Mandarin, were conducted with physicians, community leaders, and first generation members of the Manhattan Chinatown community. Participants were asked to discuss their beliefs about cancer, cancer screening, clinical trials, and cancer health education materials. Focus group data were stratified by respondent group and analyzed for thematic content. Eleven physicians, 15 community leaders, and 38 community members participated. Some community members were not familiar with cancer screening as a preventive measure and had not received common screens such as PAP smears or mammograms. They described widespread misconceptions about cancer that act as screening deterrents, e.g. testing for cancer can cause cancer. Community members were unfamiliar with clinical trials and would not participate in a clinical trial unless "sick," and only on the recommendation of their physicians. Physicians did not see the relevance or value of clinical trials for their patients. Among first generation Chinese immigrants, there are many perceptual barriers to cancer screening and clinical trials recruitment. There is a need for effective culturally tailored health education on these health topics to address persistent misconceptions about cancer and to increase knowledge about cancer screening and clinical trials. Health education efforts and clinical trial recruitment in this community must involve community physicians. PMID- 16370056 TI - Hepatitis B awareness, testing, and knowledge among Vietnamese American men and women. AB - Southeast Asians have higher rates of liver cancer than any other racial/ethnic group in the United States. Chronic carriage of hepatitis B virus (HBV) is the most common underlying cause of liver cancer in the majority of Asian populations. Our objectives were to describe Vietnamese Americans' awareness of hepatitis B, levels of HBV testing, and knowledge about hepatitis B transmission; and to compare the HBV knowledge and practices of men and women. A community based, in-person survey of Vietnamese men and women was conducted in Seattle during 2002. Seven hundred and fifteen individuals (345 men and 370 women) completed the questionnaire. Eighty-one percent of the respondents had heard of hepatitis B (76% of men, 86% of women) and 67% reported HBV testing (66% of men, 68% of women). A majority of the participants knew that HBV can be transmitted during sexual intercourse (71% of men, 68% of women), by sharing toothbrushes (67% of men, 77% of women), and by sharing razors (59% of men, 67% of women). Less than one-half knew that hepatitis B is not spread by eating food prepared by an infected person (46% of men, 27% of women), nor by coughing (39% of men, 25% of women). One-third of our respondents did not recall being tested for HBV. Important knowledge deficits about routes of hepatitis B transmission were identified. Continued efforts should be made to develop and implement hepatitis B educational campaigns for Vietnamese immigrant communities. These efforts might be tailored to male and female audiences. PMID- 16370057 TI - Monitoring of global cerebral ischemia using wavelet entropy rate of change. AB - In this paper, the subband wavelet entropy (SWE) and its time difference are proposed as two quantitative measures for analyzing and segmenting the electroencephalographic (EEG) signals. SWE for EEG subbands, namely Delta, Theta, Alpha, Beta, and Gamma, is calculated and segmented using wavelet analysis. In addition, a time difference entropy measure was calculated because it does not require a baseline and equals to zero in all clinical bands as the initial condition. Visual and quantitative results were obtained from 11 rodents that were subjected to 3, 5, and 7 min of global ischemic brain injury by asphyxic cardiac arrest. We found that the time difference of SWE is capable of amplifying the variations between clinical bands during the various stages of the recovery process and may serve as a novel analytical approach to grade and classify brain rhythms during global ischemic brain injury and recovery. PMID- 16370058 TI - Poly(ether urethane)s incorporating long alkyl side-chains with terminal carboxyl groups as fatty acid mimics: synthesis, structural characterization and protein adsorption. AB - The object of this work was to produce polyurethanes with greater affinity for albumin (Alb) and improved hemocompatibility by introduction of carboxyl terminated alkyl side-chains that better mimic fatty acids, in contrast to methyl terminated alkyl side-chains used previously. Synthesis of poly(ether urethane)s (PEUs) with long alkyl side-chains via a multi-step solution addition polymerization is described. The synthesis is based upon the polymerization of a diisocyanate pre-polymer with various chain extenders and reaction with Br terminated compound in the final stage. The side-chains had terminal methyl or carboxylic groups, and were attached either directly to the polymer backbone or to an oligo(ethylene glycol) spacer. The bulk structure of the PEUs was confirmed by 1H-NMR and the surface polymer structure was characterized by ToF-SIMS. The influence of the incorporated C16-alkyl, C16-carboxyalkyl and oxyethylene-C16 carboxyalkyl side-chains attached to the polymer backbone on fibrinogen (Fg) and Alb adsorption from blood plasma, and Fg adsorption from buffer solutions and binary mixtures with Alb was measured. Incorporation of C16-alkyl or C16 carboxyalkyl side-chains into PEUs caused relatively small changes in Fg and Alb adsorption. PEUs with oxyethylene-C16-carboxyalkyl side-chains exhibited the lowest Fg adsorption and the highest Alb adsorption among all the tested polymers. PMID- 16370059 TI - Improved phenotype of rat islets in a macrocapsule by co-encapsulation with cross linked Hb. AB - A number of rat islets were co-encapsulated in a diffusion chamber-type device, i.e., macrocapsule, with a thermoreversible polymeric extracellular matrix (ECM) and bioactive ingredient of cross-linked hemoglobin (Hb-C). The ECM was formed from an aqueous solution of N-isopropyl-acrylamide co-polymers with a small amount of acrylic acid, which exhibited unique sol-gel transition in a temperature range of 30-34 degrees C, without noticeable hysteresis. The incorporation of Hb-C in the islet macrocapsule showed a concentration-dependent effect on insulin secretion and viability of the entrapped islets. Insulin secretion stimulation by glucose and cell viability were more than doubled when compared with a control group (without Hb-C), at an optimum Hb-C concentration of 0.25 mM due to its unique oxygen transporting capacity. Furthermore, 0.25 mM Hb-C in the macrocapsule was able to support islet density up to 1000 islets/device in a 154 microl total volume without negative effects on islet functionality and viability. Hb-C incorporation is an effective strategy for a macrocapsule-type biohybrid artificial pancreas for Type-I diabetes treatment, which can be further developed to a rechargeable system by employing the thermoreversible ECM and designing a proper macrocapsule. PMID- 16370060 TI - Who rules the great Indian drug bazaar? PMID- 16370061 TI - Needed: closer scrutiny of clinical trials. PMID- 16370062 TI - Can the medical profession and the pharmaceutical industry work ethically for better health care? PMID- 16370063 TI - Unhealthy drug donations. PMID- 16370064 TI - A comparison of codes of pharmaceutical marketing practices. PMID- 16370065 TI - It's time. AB - Statistical inference involves taking the results of models and knowledge about probability to make decisions about the relationship in question. This commentary explains the usefulness of statistical inference to the drug development process, as well as some common pitfalls. It also examines reasons why statistical inference does not seem to be fully integrated into pharmacometric modeling. An example is shown that demonstrates the inferential advantages of mechanistic models. Both statisticians and pharmacometricians ought to take note of these advantages and integrate their efforts in order to maximize the decision-making potential of clinical research. PMID- 16370066 TI - Ethical considerations in laparoscopic surgery. PMID- 16370067 TI - Doctors, drug companies and medical ethics: a Sri Lankan perspective. PMID- 16370068 TI - Informed consent--a view from the trenches. PMID- 16370069 TI - What shoulder and what art. PMID- 16370070 TI - Star role for bacteria in controlling flu pandemic? PMID- 16370071 TI - The kidney trade again. PMID- 16370072 TI - First phase of HapMap project already helping drug discovery. PMID- 16370073 TI - Government-funded anti-retroviral therapy for HIV/AIDS: new ethical challenges. PMID- 16370074 TI - Medical treatment inventions. PMID- 16370075 TI - An audience with... Fred Hassan. PMID- 16370076 TI - Access to AIDS medicine: ethical considerations. PMID- 16370077 TI - Why life-saving drugs should be public goals. PMID- 16370078 TI - All PPIs equivalent for treatment of GERD. PMID- 16370079 TI - Newer antipsychotics similar to older agents. PMID- 16370080 TI - Authorized generics. PMID- 16370082 TI - Shared decision-making not for all patients. PMID- 16370081 TI - Headaches associated with OCs tend to improve over time. PMID- 16370083 TI - Measuring blood pressure at home is valuable. PMID- 16370084 TI - Ensuring quality of care in sterilisation services. PMID- 16370085 TI - Ethical issues in psychiatry. PMID- 16370086 TI - Tipranavir. PMID- 16370087 TI - Unmet ethical concerns of the proposed preventive HIV vaccine trials in India. PMID- 16370088 TI - Euthanasia: a worldwide dilemma. PMID- 16370089 TI - Publication ethics policies for medical journals. PMID- 16370090 TI - The Helsinki Declaration, 2000, and ethics of human research in developing countries. PMID- 16370091 TI - Assay development: an increasingly creative endeavour. PMID- 16370092 TI - Annotated bibliography of NINR findings on women's health across the life span: 2005 update. PMID- 16370093 TI - Symposium summary. PMID- 16370094 TI - Abstracts from the Metabolic Syndrome, Type II Diabetes, and Atherosclerosis Congress. May 19-23, 2004. Marrakech, Morocco. PMID- 16370095 TI - Strategies for optimizing management of menstrual migraine. PMID- 16370096 TI - Diabetes type and endogenous insulin. PMID- 16370097 TI - Tegaserod for chronic constipation. PMID- 16370098 TI - Family medicine: aiming higher. PMID- 16370099 TI - COPD and antibiotics. PMID- 16370100 TI - Medicalisation of 'legal' killing: doctors' participation in the death penalty. PMID- 16370101 TI - Addressing ethical concerns in the Indian HIV vaccine trials. PMID- 16370102 TI - Intellectual capital as property. PMID- 16370103 TI - Provision of health care by the government. PMID- 16370104 TI - Concerns about ethical review of health research in India. PMID- 16370105 TI - Role of ethics committees in medical research. PMID- 16370106 TI - Research in a tribal community. PMID- 16370107 TI - Purity of standards: at what price? PMID- 16370108 TI - Challenges in research in tribal communities. PMID- 16370109 TI - Withdrawal from dialysis: ethical issues. AB - Since 1991, death following withdrawal from dialysis has increased greatly in our dialysis unit. This report is based on our observations of those patients who followed that course. Four types of patients who withdrew from dialysis were identified: those with a terminal illness, demented patients, those with a progressive disability, and those who had no serious medical problem other than end-stage renal failure. We analyzed the risk factors for withdrawal and attempted to define the ethical principles involved in each patient category. The authors conclude that although the decision of a competent patient to stop dialysis must be honored, some of those deaths might be preventable if patients on chronic dialysis are prospectively followed and treated by those who are expert in the behavior of patients with chronic illness. PMID- 16370110 TI - Evolutionary significance of promiscuity in an aphidophagous ladybird, Propylea dissecta (Coleoptera: Coccinellidae). AB - The short- and long-term consequences of monogamy and promiscuity in an aphidophagous ladybird, Propylea dissecta (Mulsant) in terms of reproductive performance and offspring fitness were investigated. Promiscuous females were more fecund and laid more viable eggs than monogamous ones. Amongst promiscuous females, those mated with multiple males (freedom for mate choice) had significantly higher reproductive output than those mated daily with a new unmated male. The increased reproductive output of females mated with an unmated male each day in comparison to monogamous ones might possibly be a result of their increased ejaculate size. The further increase in reproductive output, when a female was allowed to choose a mate, may be because of repeated preferential mating with the fitter males. Results on long-term consequences of promiscuity revealed faster development and increased survival in offspring of promiscuous females at 25, 27 and 30 degrees C. The percent adult emergence was more strongly influenced than development by the mating regime. There were no statistically significant differences in survival of offspring of promiscuous females (mate choice) at the three temperatures, while the percent survival of offspring of monogamous females differed significantly. Offspring of promiscuous females (mate choice) were fittest in terms of development and survival, which indicates their probable better genetic constitution and subsequently that of chosen mates. Thus, promiscuity has both short- and long-term benefits and is advantageous to ladybirds. The presence of long-term benefits in terms of fitter offspring probably indicates the evolutionary rationale behind the prevalence of promiscuity in ladybirds. PMID- 16370111 TI - Comment on: J Gimnig et al. (2005) Laboratory wash resistance of long-lasting insecticidal nets. Tropical Medicine & International Health 10, 1022-9. PMID- 16370112 TI - The biogeochemical cycles of the elements and the evolution of life. PMID- 16370113 TI - Biogeochemistry of dihydrogen (H2). AB - Hydrogen has had an important and evolving role in Earth's geo- and biogeochemistry, from prebiotic to modern times. On the earliest Earth, abiotic sources of H2 were likely stronger than in the present. Volcanic out-gassing and hydrothermal circulation probably occurred at several times the modern rate, due to presumably higher heat flux. The H2 component of volcanic emissions was likely buffered close to the modern value by an approximately constant mantle oxidation state since 3.9 billion years ago, and may have been higher before that, if the early mantle was more reducing. The predominantly ultramafic character of the early, undifferentiated crust could have led to increased serpentinization and release of H2 by hydrothermal circulation, as in modern ultramafic-hosted vents. At the same time, the reactive atmospheric sink for H2 was likely weaker. Collectively, these factors suggest that steady state levels of H2 in the prebiotic atmosphere were 3-4 orders of magnitude higher than at present, and possibly higher still during transient periods following the delivery of Fe and Ni by large impact events. These elevated levels had direct or indirect impacts on the redox state of the atmosphere, the radiation budget, the production of aerosol hazes, and the genesis of biochemical precursor compounds. The early abiotic cycling of H2 helped to establish the environmental and chemical context for the origins of life on Earth. The potential for H2 to serve as a source of energy and reducing power, and to afford a means of energy storage by the establishment of proton gradients, could have afforded it a highly utilitarian role in the earliest metabolic chemistry. Some origin of life theories suggest the involvement of H2 in the first energy-generating metabolism, and the widespread and deeply-branching nature of H2-utilization in the modern tree of life suggests that it was at least a very early biochemical innovation. The abiotic production of H2 via several mechanisms of water-rock interaction could have supported an early chemosynthetic biosphere. Such processes offer the continued potential for a deep, rock-hosted biosphere on Earth or other bodies in the solar system. The continued evolution of metabolic and community-level versatility among microbes led to an expanded ability to completely exploit the energy available in complex organic matter. Under the anoxic conditions that prevailed on the early Earth, this was accomplished through the linked and sequential action of several metabolic classes of organisms. By transporting electrons between cells, H2 provides a means of linking the activities of these organisms into a highly functional and interactive network. At the same time, H2 concentrations exert a powerful thermodynamic control on many aspects of metabolism and biogeochemical function in these systems. Anaerobic communities based on the consumption of organic matter continue to play an important role in global biogeochemistry even into the present day. As the principal arbiters of chemistry in most aquatic sediments and animal digestive systems, these microbes affect the redox and trace-gas chemistry of our oceans and atmosphere, and constitute the ultimate biological filter on material passing into the rock record. It is in such communities that the significance of H2 in mediating biogeochemical function is most strongly expressed. The advent of phototrophic metabolism added another layer of complexity to microbial communities, and to the role of H2 therein. Anoxygenic and oxygenic phototrophs retained and expanded on the utilization of H2 in metabolic processes. Both groups produce and consume H2 through a variety of mechanisms. In the natural world, phototrophic organisms are often closely juxtaposed with a variety of other metabolic types, through the formation of biofilms and microbial mats. In the few examples studied, phototrophs contribute an often swamping term to the H2 economy of these communities, with important implications for their overall function-including regulation of the redox state of gaseous products, and direct release of large quantities of H2 to the environment. As one of the dominant sources of biological productivity for as much as 2 billion years of Earth's history, these communities have been among the most important agents of long-term global biogeochemical change. On the modern Earth, H2 is present at only trace levels in the atmosphere and oceans. Nonetheless, its function as an arbiter of microbial interactions and chemistry ensures an important role in biogeochemical cycling. The significance of H2 in a global sense may soon increase, as the search for alternative fuels casts attention on the clean-energy potential of hydrogen fuel cells. Already, H2 utilization plays an important role in all three phylogenetic domains of life. Humans may soon add an important new term to this economy. Considerable research is focused on the H2-producing capacities of phototrophic and other microorganisms as potential contributors in this regard. Regardless of source, the large scale utilization of H2 as an energy source could carry important consequences for biogeochemistry. PMID- 16370114 TI - Dioxygen over geological time. PMID- 16370115 TI - The nitrogen cycle: its biology. PMID- 16370116 TI - The biological cycle of sulfur. PMID- 16370117 TI - Biological cycling of phosphorus. PMID- 16370118 TI - Iron, phytoplankton growth, and the carbon cycle. AB - Iron is an essential nutrient for all living organisms. Iron is required for the synthesis of chlorophyll and of several photosynthetic electron transport proteins and for the reduction of CO2, SO4(2-), and NO3(-) during the photosynthetic production of organic compounds. Iron concentrations in vast areas of the ocean are very low (<1 nM) due to the low solubility of iron in oxic seawater. Low iron concentrations have been shown to limit primary production rates, biomass accumulation, and ecosystem structure in a variety of open-ocean environments, including the equatorial Pacific, the subarctic Pacific and the Southern Ocean and even in some coastal areas. Oceanic primary production, the transfer of carbon dioxide into organic carbon by photosynthetic plankton (phytoplankton), is one process by which atmospheric CO2 can be transferred to the deep ocean and sequestered for long periods of time. Accordingly, iron limitation of primary producers likely plays a major role in the global carbon cycle. It has been suggested that variations in oceanic primary productivity, spurred by changes in the deposition of iron in atmospheric dust, control atmospheric CO2 concentrations, and hence global climate, over glacial interglacial timescales. A contemporary application of this "iron hypothesis" promotes the large-scale iron fertilization of ocean regions as a means of enhancing the ability of the ocean to store anthropogenic CO2 and mitigate 21st century climate change. Recent in situ iron enrichment experiments in the HNLC regions, however, cast doubt on the efficacy and advisability of iron fertilization schemes. The experiments have confirmed the role of iron in regulating primary productivity, but resulted in only small carbon export fluxes to the depths necessary for long-term sequestration. Above all, these experiments and other studies of iron biogeochemistry over the last two decades have begun to illustrate the great complexity of the ocean system. Attempts to engineer this system are likely to provoke a similarly complex, unpredictable response. PMID- 16370119 TI - The biogeochemistry of cadmium. PMID- 16370120 TI - The biogeochemistry and fate of mercury in the environment. PMID- 16370121 TI - Biogeochemistry and cycling of lead. AB - Lead has no biological function and is one of the most toxic metals. At the same time, it is one of the most useful, and perhaps no other metal has found such a wide range of industrial applications. It has been used extensively since Antiquity, which is when environmental Pb contamination began. With respect to contamination since industrialization, peat bogs and polar ice show that coal combustion and industrial emissions were as important during the first half of the 20th century as gasoline lead was during the second half. Air Pb concentrations have generally declined since the introduction of pollution control technologies and the gradual elimination of leaded gasoline additives. However, all of the most recent published studies of the isotopic composition of Pb in aerosols and archival samples show that anthropogenic sources continue to dominate the atmospheric Pb flux by a considerable margin. The health effects of childhood Pb exposure is a growing concern, as deleterious effects are seen at BLL well below those currently believed to be safe, and safe levels are one or two orders of magnitude above the estimated natural values. Mobilization of Pb rich particles from highly contaminated soils in urban areas is an on-going health concern for many large cities. Even in areas far removed from industrial emission sources, Pb concentrations in the surface soil layers are far above their natural concentration range. In acidic forest soils, Pb concentrations are not only elevated in the biologically active zone, but also in their corresponding pore fluids. Accurate and precise measurements of the isotopic composition of Pb employing appropriate clean lab protocols, will continue to advance our understanding of the fate of Pb in the environment and its impact on human and ecosystem health. PMID- 16370122 TI - Interview with Chip Holmes, CHE, administrator, Littleton Regional Hospital, Bethlehem, New Hampshire. Interview by Kyle L Grazier. PMID- 16370123 TI - Patterns of financing for the largest hospital systems in the United States. AB - The ten large systems reviewed in this column have greater degrees of financial leverage than do most freestanding hospitals. Larger firms typically have both greater capital access and lower costs of financing. Both voluntary and IO systems make extensive use of variable rate financing, but the percentage of variable rate financing is slightly higher for voluntary systems. This difference may be attributable to larger yield curve spreads for tax-exempt versus taxable securities. Interest rate swaps were used by 70 percent of the systems, but the actual amount swapped was relatively minor. This may change in the future as financial officers become more comfortable and familiar with interest rate swap arrangements. When compared to IO systems, voluntary systems have extensive levels of cash relative to their debt positions. Cash balances are more critical in the bond-rating process for voluntary hospitals, and the ability to raise new equity is much more limited in the voluntary sector. Very little capital leasing was used in any of the systems. PMID- 16370124 TI - Dealing with "difficult" people. PMID- 16370125 TI - Self-assessment of cultural and linguistic competence in an ambulatory health system. AB - The University Health System (UHS) leaders organized a comprehensive planning process of its ambulatory care system. As part of this planning process, UHS assessed the cultural and linguistic competence of its outpatient environment. This assessment was conducted within the context of standards outlined in the U.S. Department of Health & Human Services (2001) Standards for Cultural and Linguistically Appropriate Services, using a multimethod approach. This multimethod approach included (1) a review of the organizational profile, (2) a review of organizational documents, (3) a review of policies and procedures, (4) clinic site visits, (5) a staff survey, (6) patient and provider focus groups, and (7) interviews with key stakeholders in the community. Generally, the UHS was found to have several strengths that enable it to continue meeting the medical needs of its target populations. To develop greater capacity in the areas of cultural and linguistic competencies, the UHS should continue to promote respect for diversity by focusing on attitudes; beliefs; behaviors; practices; and communication patterns associated with race, ethnicity, religion, socioeconomic status, historical and social context, physical or mental ability, age, gender, sexual orientation, generational status, and acculturation level. Recommendations for achieving a more culturally competent healthcare organization are provided. PMID- 16370126 TI - Reinventing Veterans Health Administration: focus on primary care. AB - Can we improve access in primary care without compromising the quality of care? The purpose of this article is to demonstrate how timely access to primary care can be achieved without compromising the quality of the care being delivered. The Veterans Health Administration (VHA) is an integrated healthcare system that has implemented change to improve primary care access to the veterans it serves, while not only maintaining but also actually improving the quality of care. Many healthcare executives are struggling with achieving desirable access to care and continuity of care. To confront this problem, many large and small practices have initiated an approach known as advanced clinic access, open access, or same-day scheduling, introduced by the Institute for Healthcare Improvement (IHI). This approach has increasingly been used to reduce waits and delays in primary care without adding resources. To measure quality of care, specific performance measures were developed to quantify the effectiveness of primary care in VHA. Although it was initially viewed with concern and suspicion and was seen as a symptom of unnecessary micromanagement, healthcare team members were encouraged to use performance feedback as an opportunity for systems improvement as well as self-assessment and performance improvement for the team. All quality data are posted quarterly on VHA's internal web site, providing visible accountability at all levels of the organization. Clinical workflow redesign leads to reduced wait times without compromising quality of care. These large system improvements are applicable to large and small organizations looking to tackle change through the use of a collaborative model. PMID- 16370127 TI - Design and implementation of a comprehensive heart failure management program. PMID- 16370128 TI - From synapse to psychotherapy: the fascinating evolution of neuroscience. AB - This paper reviews neuroscientific advances across the therapeutic disciplines and outlines new potential insights into understanding the biology of the psychotherapeutic processes. In a quest to understand the modus operandi of psychotherapy, the author reviews the evolution of the biology of psychotherapy. The mechanisms involved in learning processes, such as memory and priming, attachment, and long-term consequences of early life trauma, demonstrate how brain structures can be affected by environmental changes. Finally, the author proposes that verbal and non-verbal aspects of the psychotherapeutic process mediate their therapeutic effects through biological changes as they work on primitive emotional reflexes and stimulate mentalization processes. PMID- 16370129 TI - Therapist self-disclosure to child patients. AB - The primary aim of this study was to investigate therapists' use of self disclosure with their child patients. A sample of 126 mental health professionals with an average of 20 years of clinical experience completed the Therapist-to Child Disclosure Inventory (TCDI), a 42-item Likert-type measure created for this study. Therapist self-disclosure was examined using three principle dimensions: the mean frequency of specific disclosures, the degree to which child patients solicit these disclosures, and the extent to which specific self-disclosures are seen as advancing treatment aims for child patients. Results indicate that therapists reveal personal information to their child patients infrequently, that children almost never solicit personal disclosures from their therapists, and that therapists perceive self-disclosure as seldom advancing treatment aims for child therapy. Future studies, the authors suggest, should examine the differences between therapists' use of self-involving disclosures to children and their use of factual self-disclosures. PMID- 16370130 TI - The clinical relevance of psychophysiology: support for the psychobiology of empathy and psychodynamic process. AB - Psychophysiologic measures, such as skin conductance and heart rate, have been used in both psychotherapy process research and clinical practice. We present a case report of a patient and therapist who participated in a process-oriented psychotherapy research protocol using simultaneous measures of skin conductance. Data from the research protocol were used to broaden an empathic understanding of the patient, which facilitated insight and enhanced the exploration of conscious and unconscious processes that originated in the past and have come to dominate the present--the core of psychodynamic theories of change. The case illustrates the clinical relevance of psychophysiology and its use as a potential bridge between psychotherapy research and the theory and practice of psychotherapy. The implications of the case in support of the role of empathy in psychotherapy are discussed. PMID- 16370131 TI - Psychotherapy with intercultural couples: a contemporary psychodynamic approach. AB - Although the number of intercultural marriages has dramatically increased in the last three decades, few articles have been published on the counseling of intercultural couples. The authors propose a methodology for working with these couples using intersubjectivity and self-psychology as theoretical perspectives. The role of culture in the construction of the individual's subjectivity is seen as central to understanding the psychodynamics of the intercultural couple. Two case studies are presented in order to highlight the theoretical assumptions underlying the authors' perspectives. PMID- 16370132 TI - Impoverished dialogical relationship patterns in paranoid personality disorder. AB - In the opinion of many experts, the self is made up of numerous different, independent facets interacting with each other in an ongoing inner dialogue. The meaning of events depends on the form this dialogue takes. The hypothesis we discuss in this article is that patients suffering from paranoid personality disorder (PPD) present impoverished dialogical relationship patterns. By this we mean that: a) The characters operating on their mental stage are few and repetitive. The character identified as self is insufficient-inadequate or diffident-mistrusting-hostile. The characters embodied by other persons are hostile, humiliating, and threatening. b) The inner dialogue the characters set up is stereotyped and always has the same outcome--the inadequate part of self feels under attack by a hostile other. This pattern has an influence on patients' behaviour and the course of psychotherapy. Our discussion of this hypothesis will be based on an analysis of extracts from diaries written by a patient with PPD during therapy. We shall give a number of strategies as to how a therapist may avoid patient drop-outs and provide effective treatment. PMID- 16370133 TI - Adjusting cognitive behavior therapy for adolescents with bulimia nervosa: results of case series. AB - This article reviews the types of adjustments needed to an adult protocol of cognitive-behavioral therapy (CBT) for bulimia nervosa (BN) to make it more acceptable to an adolescent population. Employing developmental principles as well as clinical experience as guidelines, these modifications include the involvement of parents, recognition of the interaction of treatment with normal adolescent developmental tasks, and allowances for typical cognitive and emotional immaturity on treatment procedures. Outcomes from a series of adolescents with BN who were treated with this modified-CBT approach show results similar to those expected in adult populations treated using CBT. PMID- 16370134 TI - Basic physiology of the growth hormone/insulin-like growth factor axis. PMID- 16370135 TI - Experimental models for understanding the role of insulin-like growth factor-I and its receptor during development. PMID- 16370136 TI - Insulin-like growth factor binding proteins in development. AB - IGFBPs regulate growth and development by regulating IGF transport to tissues and IGF bioavailability to IGF receptors at cell membrane level. IGFBP excess leads predominantly to inhibition of IGF action and growth retardation with impaired organogenesis. Absence of human and also mouse ALS leads to decreased IGF-I levels in circulation and causes mild growth retardation. Although IGFBP KO mice demonstrate relatively minor phenotypes, the possibility of compensatory mechanisms that mask the phenotypic manifestation of lack of individual binding proteins needs to be further investigated. Recent studies of hepatic regeneration in IGFBP-1 KO mice and also with mutant IGFBP-3 Tg mice provide some limited support for the existence of IGF-independent mechanism of action in vivo. PMID- 16370137 TI - Growth hormone, insulin-like growth factors and the female reproductive system. PMID- 16370138 TI - Skeletal muscle. PMID- 16370139 TI - Cardiovascular and respiratory systems. PMID- 16370140 TI - Hematopoiesis. PMID- 16370141 TI - Regulation of vertebrate sensory organ development: a scenario for growth hormone and insulin-like growth factors action. PMID- 16370142 TI - Role of insulin-like growth factors in neuronal plasticity and neuroprotection. PMID- 16370143 TI - Aging and life span. PMID- 16370144 TI - Interactions of insulin-like growth factor-I and estrogen in the brain. PMID- 16370145 TI - Genetic basis of proportional short stature. PMID- 16370146 TI - Future perspectives: from stem cells and IGF biology to the clinic. PMID- 16370147 TI - Adaptive on-line classification for EEG-based brain computer interfaces with AAR parameters and band power estimates. AB - We present the result of on-line feedback Brain Computer Interface experiments using adaptive and non-adaptive feature extraction methods with an on-line adaptive classifier based on Quadratic Discriminant Analysis. Experiments were performed with 12 naive subjects, feedback was provided from the first moment and no training sessions were needed. Experiments run in three different days with each subject. Six of them received feedback with Adaptive Autoregressive parameters and the rest with logarithmic Band Power estimates. The study was done using single trial analysis of each of the sessions and the value of the Error Rate and the Mutual Information of the classification were used to discuss the results. Finally, it was shown that even subjects starting with a low performance were able to control the system in a few hours: and contrary to previous results no differences between AAR and BP estimates were found. PMID- 16370148 TI - [Retransfusion of unwashed drainage blood after total hip and knee arthroplasty]. AB - The method of retransfusion of drainage blood as known from the literature was investigated in a prospectiv study regarding effectivness and rate of side effects. 200 patients who underwent total hip and knee arthroplasty were investigated concerning hemoglobin, hematocrit, amount and quality of the retransfused drainage blood, the amount of autologous and homologous transfusions as well as complications and costs. 100 of these patients were selected as the control group. The amount of the retransfused drainage blood after hip arthroplasty amounted an average of 387 +/- 194 ml and after knee arthroplasty 595 +/- 250 ml. The retransfused blood had an average hemoglobin of 5,2 +/- 0,9mmol/l with a hematocrit of 0,24 0,05. No complications directly associated to the retransfusion were found. The need of transfusion was reduced for patients with knee arthroplasty about 30% and for hip arthroplasty about 25%. The retransfusion of unwashed drainage blood is a sufficient method to reduce perioperative homologous blood transfusion in patients with arthroplasty of hip and knee. Substantial complications were not observed, so that this method seems to be save enough for clinical usage. The method is easy to handle and usable without special technical devices. The autologous retransfusion of drainage blood can contribute to lower costs in patients treatement. PMID- 16370149 TI - Effects of the rib cage on thoracic spine flexibility. AB - Besides protecting the internal organs of the thorax, the rib cage is the site of numerous muscle attachments. It also decreases the overall flexibility of the thoracic spine. This study developed finite element (FE) models of the thoracic spine with and without the rib cage, and the effects of the rib cage on thoracic spine flexibility were determined. The numerical models were validated by comparing the maximum rotation of the models for several loading cases with experimental data in the literature. After adapting the material properties for the discs and ligaments, the calculated maximum rotations differed from the measured median values by less than 1 degrees without the rib cage and by less than 2.5 degrees with it. The rib cage decreased the mean flexibility of the thoracic spine by 23% to 47%, depending on the loading plane. Assuming the ribs to be rigid beams required a corresponding reduction of ligament stiffnesses in order to achieve the same agreement of the maximum rotations with the measured median values. Interconnecting the FE thoracic spine model plus rib cage with the existing detailed FE lumbar spine model improves the simulation of force directions of muscles attached to the rib cage or thoracolumbar spine. In addition, such a model is suitable for determining the effects of lumbar spine implants on spinal balance. PMID- 16370150 TI - [The influence of the initial humidity of balneological peat on its pharmacological features]. AB - The viscosity of balneological peat got special meaning according to its thermical features. Until now, there has been no answer on the question in how far balneological peat, which is prepared with different degrees of humidity, can vary the pharmacological character. Aim of this work was, on the one hand, to examine which initial degree of peat-humidity is necessary to get the wished viscosity in therapy when diluted with water. On the other hand, how far the pharmacological character of balneological is influenced by the initial degree of humidity. From peat, with initial different degrees of humidity, balneological peat is prepared, fulfilling the demands of the Quentin-Test and therefore the necessary consistence for therapy. Here in this work the in-vitro effect of aqueous peat extract (APE) on the spontaneous contractile activity of the smooth muscle fibres of guinea-pig stomachs was examined. The APE used got initial different degrees of humidity. It was found out, that the ability of peat according to a new water-uptake, with a initial humidity under 60%, was clearly reduced. Because of the reduced water content of balneological peat its thermical features were deteriorated. This goes for peat with a humidity factor < 60%. First, with the Quentin-Test, showing the optimal viscosity of peat before medical use, you can also make statements according its ability of maximum uptake of water, meaning the ability taking up water until it becomes gel-like. Second, the results show that the initial humidity of peat got no influence on the pharmacological features. PMID- 16370151 TI - [German Artificial Sphincter System--GASSII: first in vivo evaluation of a novel and highly integrated sphincter prosthesis for therapy of major fecale incontinence]. AB - The German Artificial Sphincter System GASS consists of a support ring which includes a fluid reservoir on the outer side and an occlusive cuff on the inner side. The cuffs are designed as polyurethane hollow bodies with a pre-determined inflation volume and are connected to an integrated piezo micropump/valve unit. To evaluate the threshold of continence, the GASS was placed around the anorectal junction via a perineal approach in one mini pig. The novel cuff design reduces the occlusion pressure and allows low compression volumes. Low operating pressures indicate a minor risk of ischemia injury of the bowel. The operation time is estimated at about 6 days with no recharging of the battery. The novel remote controlled GASS is a highly integrated prosthesis for placement around the anal canal or lower rectum and is effective in restoring continence for liquids and solids in vitro and in vivo. PMID- 16370152 TI - Roles of and correlation between alpha-smooth muscle actin, CD44, hyaluronic acid and osteopontin in crescent formation in human glomerulonephritis. AB - BACKGROUND: alpha-Smooth muscle actin (SMA), CD44, hyaluronic acid (HA) and osteopontin (OPN) are involved in crescent formation; however, the correlation between these molecules during the formation and progression of the crescents in human glomerulonephritis (GN) has not been fully evaluated. METHODS: To investigate the expression of alpha-SMA, CD44, HA, OPN and CD68 renal biopsy specimens from 14 patients with crescentic GN were examined by immunohistochemistry. All crescents were separated into cellular, fibrocellular and fibrous. The extent of staining in each crescent was scored semiquantitatively. The change in the expression of each molecule and its correlation with other molecules during the formation and progression of the crescents were estimated statistically. RESULTS: The expression of alpha-SMA was significantly up-regulated in the fibrocellular crescents compared with that in the cellular and fibrous crescents. The expression of CD44, OPN and CD68 was significant in the cellular crescents compared with that in the fibrocellular and fibrous crescents. The deposition of HA in the three groups of crescents was high level. However, that of HA was not significant among three groups of crescent. The expression of CD44 in the cellular crescents correlated significantly with the expression of OPN and CD68, and the deposition of HA in the cellular crescents. The expression of OPN in the cellular crescents correlated with the deposition of HA and the expression of CD68 in the cellular crescents. The expression of alpha-SMA in the cellular and fibrocellular crescents correlated with the deposition of HA in the cellular and fibrocellular crescents. CONCLUSION: The expression of CD44, HA, OPN and CD68 was up-regulated at the early stage of the crescent formation in human crescentic GN. Moreover, myofibroblasts and cell-matrix interactions mediated by the CD44-OPN and CD44-HA receptor-ligand pairs may play important roles in the formation and progression of the crescents. PMID- 16370153 TI - Risk of bleeding and restenosis among chronic kidney disease patients undergoing percutaneous coronary intervention. AB - BACKGROUND: Bleeding risk is increased in renal failure due to impaired platelet adhesiveness. Patients who undergo percutaneous coronary intervention (PCI) are given multiple antiplatelet agents that increase that risk. We retrospectively tested the hypothesis that chronic kidney disease (CKD) patients who undergo PCI are at higher risk of bleeding and restenosis (due to chronic inflammation) compared to patients with normal renal function. METHODS: Patients who had PCI for non-ST elevation myocardial infarction or unstable angina between July 2001 and June 2003 (1,184 patients) were included in the study. All the patients were given periprocedural clopidogrel, aspirin and glycoprotein IIb/IIIa inhibitor if indicated, and then continued on clopidogrel and aspirin daily for 12 months. The patients were classified into 5 groups according to the CKD stage and followed-up for 12 months for development of major or minor bleeding, restenosis, length of hospital stay and survival. RESULTS: The incidence of major bleeding within the first month (3.4% in normal kidney function patients (Gp 1), 4.8% for CKD Stages 1 and 2 patients (Gp2), 5.2% for CKD Stage 3 patients (Gp3), 6.1% for CKD Stage 4 patients (Gp4) and 9.3% for CKD Stage 5 patients (Gp5), p = 0.001) and for minor bleeding (5.7% in Gp1, 6.5% for Gp2, 7.4% for Gp3, 9.2% for Gp4 and 11.3% for Gp5, p = 0.001) and the incidence of restenosis at one month (4.6% in Gp1, 5.3% for Gp2, 6.8% for Gp3, 7.3% for Gp4 and 9.6% for Gp5, p = 0.001) and 6 months (11.2% in Gp1, 13.5% for Gp2, 15.7% for Gp3, 16.4% for Gp4 and 19.7% for Gp5, p = 0.001) were higher with worsening CKD. Survival at one year was worse with worsening of the kidney function. CONCLUSION: Worsening of CKD is associated with progressively increased risk of minor and major bleeding, restenosis and death during and after PCI. PMID- 16370154 TI - Relations of inflammatory markers to lipid levels and autonomic tone in patients with moderate and severe chronic kidney disease and in patients under maintenance hemodialysis. AB - BACKGROUND AND AIMS: Chronic kidney disease is associated with enhanced inflammatory response and autonomic dysfunction. Evidence exists of a potential interaction of inflammation and nervous system. We sought to investigate determinants of heart rate variability (HRV) and relations between the inflammatory markers C-reactive protein (CRP) and interleukin-6 (IL-6) and autonomic tone in patients with moderate and severe chronic kidney disease and in maintenance hemodialysis patients, and relations of CRP and IL-6 with clinical characteristics and lipid levels. METHODS: This was a cross-sectional study of 51 hemodialysis and 53 moderate and severe chronic kidney disease patients. Autonomic tone was assessed using 24-hour HRV analysis in time and frequency domain. All patients underwent measurements of high sensitivity CRP, IL-6 and lipid levels. RESULTS: CRP and IL-6 were elevated in the non-dialysis group at levels similar to hemodialysis patients. Hemodialysis patients had lower total cholesterol, LDL cholesterol and apolipoprotein B levels (p < 0.05), and in this group of patients lipids were related to CRP and IL-6 (p < 0.05). The inflammatory marker IL-6 was associated to HRV in the moderate and severe chronic kidney disease group (R = -0.4, p < 0.01 for standard deviation of RR intervals and very low frequency power, R = -0.5, p < 0.01 for standard deviation of all five-minute RR intervals, R = 0.35, p < 0.05, for total power and low frequency power). Adequacy of dialysis, but not the inflammatory markers, was associated to HRV in the hemodialysis group (R = 0.6, p < 0.01 for high frequency power). CONCLUSION: Enhanced inflammatory response occurs already in stages 3 and 4 chronic kidney disease. IL-6 is related to HRV in these patients, but not in the hemodialysis group, suggesting that IL-6 may interact with autonomic tone in that stage of disease. PMID- 16370155 TI - The effects of lanthanum carbonate and calcium carbonate on bone abnormalities in patients with end-stage renal disease. AB - BACKGROUND: Renal osteodystrophy is a common complication of end-stage renal disease (ESRD) and is a major cause of morbidity in patients with ESRD. High serum levels of phosphorus, calcium and parathyroid hormone are associated with the development of this disease. The effects on bone of treatment with lanthanum carbonate, a new phosphate binder, and calcium carbonate were assessed in patients with ESRD. METHODS: This was an open-label, multicenter, parallel-group study. Patients were recruited within 12 weeks of commencing dialysis. Following screening, phosphate binder administration was stopped, tetracycline labeling administered and a transiliac bone biopsy taken. After randomization to lanthanum carbonate or calcium carbonate, patients were titrated to an optimum dose for 8 weeks and maintained at this dose for 44 weeks. The bone was then labeled and a second biopsy taken. Biopsy samples were analyzed histomorphometrically. RESULTS: Paired bone biopsies from 33 lanthanum carbonate- and 30 calcium carbonate treated patients were suitable for analysis. None of the patients on either treatment developed osteomalacia. Assessment of activation frequency changes showed that 41% of biopsies from lanthanum carbonate-treated patients moved towards normal (observed values at the follow-up biopsy were closer to expected values than were the baseline values, so patients were considered to be improved) compared with 23% of calcium carbonate-treated patients (p = 0.15). CONCLUSIONS: This study indicates that there was no evidence of aluminum-like toxicity with lanthanum carbonate after 1 year of treatment in ESRD patients commencing dialysis, and there appeared to be a beneficial effect on bone-cell function and activity compared with calcium carbonate. PMID- 16370156 TI - Comparison of transplant listing strategy in two renal dialysis centers within a regional transplant alliance. AB - AIMS: An increasing dialysis population and insufficient supply of transplant organs necessitate that patients are carefully evaluated prior to registration on the national waiting list to ensure effective utilization of a scarce resource. We have assessed listing practice in two renal units within the North of England Transplant Alliance. METHODS: Demographic, ethnic and clinical data were recorded at initiation of dialysis for patients from two northern English cities, Bradford (n = 209) and Hull (n = 202) between 1994-2000. Patients were stratified by two co-morbidity scoring systems. Multivariate and survival analyses were undertaken by registration status. RESULTS: Overall, 159 patients were registered onto the waiting list. Stratification by co-morbidity predicted listing at high and low risk, but with overlap at medium scores. There was no difference in overall co morbid burden between the two centers (p = 0.161 and 0.316, respectively, for two scoring systems). Logistic regression analysis demonstrated a center effect, Hull having an odds ratio for listing of 0.48 compared to Bradford (p = 0.041). Short- and medium-term survival in the listed group was high regardless of co-morbid score (22 vs 174 deaths in the non-listed group). In this cohort, five patients died with grafts, another three died whilst active on the waiting list. The remaining 14 patients had been removed from the list prior to death. SUMMARY: Co morbidity scoring schemes are unlikely to be sophisticated enough to accurately identify those who would most benefit from transplantation, and the value of clinical judgment is well-shown in this study. Standardization of registration will result in more equitable allocation of organs. However, this study has demonstrated that there are differences in listing practices even within a single alliance. Continuous assessment will allow judicious removal from the waiting list of patients who have developed an unacceptable co-morbid burden. PMID- 16370157 TI - Report of a case with surprising etiology of renovascular hypertension. AB - Takayasu arteritis is a chronic, idiopathic, inflammatory disease that primarily affects large vessels, such as the aorta and its main branches. Epidemiologically, it is found mostly in female patients and is more prevalent in Asian and Latin American countries. Disease may be heterogeneous in presentation. In this report, we present a different clinical expression of Takayasu arteritis in a young man who had hypertension as the sole manifestation of multiple critical arterial involvement with elevated inflammation markers but no other symptoms. A 28-year-old man was admitted with hypertension. There was no evidence for systemic vasculitis by history, serologic studies or other laboratory data. The acute-phase reactants were elevated with an erythrocyte sedimentation rate of 55 mm/h, and a C-reactive protein value of 22 mg/dl. Digital subtraction angiography showed multiple severe stenoses or occlusions of the branches of the abdominal aorta and arcus aortae together with bilateral renal artery involvement. The etiology of renovascular hypertension was found to be Takayasu arteritis with the presence of at least three criteria, as outlined by the American College of Rheumatology in 1990. Patients with Takayasu arteritis may have atypical clinical expression of the disease, and a diagnosis of Takayasu arteritis should be kept in mind in the differential diagnosis of renovascular hypertension in young subjects, even if they do not have associated symptoms of multiple arterial involvement. PMID- 16370158 TI - Laceration of gastric mucosa associated with dialysis-related amyloidosis. AB - We report a 72-year-old female on long-term hemodialysis, who was admitted to the hospital because of hematemesis. On emergency laparotomy, pylorogastrectomy was performed. The resected specimen showed a giant hematoma and traversing fissure along the lesser curvature of the body of the stomach. Histologically, the specimen showed wide hematoma formation and amyloid deposits in the submucosal layer, especially in the wall of blood vessels. These deposits reacted positively to antihuman beta2-microglobulin antibody. The post-operative course was favorable, and the patient was discharged on the 35th hospital day. In this case, the laceration site on the gastric mucosa was almost intact and did not demonstrate ischemic change, suggesting that the giant hematoma was caused by submucosal vessel rupture, which led to the gastric mucosa laceration. To our knowledge, this is the first case of gastric mucosa laceration associated with dialysis-related amyloidosis. PMID- 16370159 TI - Membrane proteinase 3 and Wegener's granulomatosis. AB - Proteinase 3 (PR3) is found in neutrophil and monocyte lysosomal granules. Anti neutrophil cytoplasmatic antibodies (ANCA) with specificity for PR3 are characteristic for patients with Wegener's granulomatosis. The interaction of ANCA with neutrophilic ANCA antigens is necessary for the development of ANCA associated diseases. ANCA bind to membrane-expressed PR3 and induce full-blown activation in primed neutrophils. We discuss two different aspects of membrane PR3 (mPR3). The first aspect is the amount of PR3 and mechanisms controlling this issue. The second aspect is the presence of two neutrophil subsets that differ in the mPR3 expression phenotype. PMID- 16370160 TI - The pathogenesis of ANCA-associated vasculitides: old hypotheses and new insights. AB - The pathomechanism of the ANCA-associated vasculitides is discussed in light of the abstracts presented at the ANCA- and Vasculitis Workshop 2005 in Heidelberg! PMID- 16370161 TI - Intravital microscopy in the study of ANCA-associated systemic vasculitis. AB - In addition to being useful clinical markers of systemic vasculitis, anti neutrophil cytoplasm antibodies (ANCA) may play a role in the initiation of vasculitic injury. These autoantibodies can induce neutrophil degranulation, dysregulated neutrophil apoptosis and neutrophil adhesion to endothelium in static cellular systems. This mini-review will place these sentinel findings in the context of more recent studies using the parallel plate flow chamber and novel animal models of ANCA-associated vasculitis (AASV). Rodent models lend themselves well to investigation of leukocyte endothelial interaction using intravital microscopy. In this way, one can study ANCA-induced leukocyte adhesion/transmigration, and microvascular injury in real time. These studies may then be extended to look at the impact of novel therapeutic agents on these processes. PMID- 16370162 TI - The role of T lymphocytes in ANCA associated vasculitis: neglected or revisited? AB - "Antineutrophil cytoplasmic antibodies (ANCA) are pathogenic--Oh yes they are!" is the title of a recent review [Falk et al. 2002], discussing the current evidence on the pathogenic role of ANCA in vasculitis. But what about T lymphocytes? Do these cells also contribute to disease manifestation and if so to what extend? T-cells most likely play a role in delivering proper signals to autoreactive B cells for the production of ANCA, but, in the efferent arm of the immune response the involvement of T cells is less obvious and controversially discussed. Numerous studies provide evidence that peripheral T-cell phenotypes are dramatically changed in ANCA associated vasculitis (AAV) patients. How these changes relate to disease manifestation is still a matter of discussion. In an attempt to provide a better understanding of how T cells might play a role in AAV, the present paper will review recent data presented at the 12th international vasculitis and ANCA workshop. PMID- 16370163 TI - Vasculitis and infection: effects of immunosuppressive therapy. AB - The immunosuppressive therapy in systemic vasculitis leads to immunological dysfunctions. The consequences of granulocytopenia and cellular immune deficits are infections of different etiologies in up to 50% of vasculitis patients. The leading severe infections are sepsis and pneumonia induced by a broad spectrum of pathogens (extra- and intra-cellular growing bacteria, fungi, parasites and viruses). The contribution of infections to the mortality of vasculitis patients is important and should induce early and careful control of these events. PMID- 16370164 TI - What is new in the therapy of ANCA-associated vasculitides? Take home messages from the 12th workshop on ANCA and systemic vasculitides. AB - ANCA-associated systemic vasculitides (AASV) were previously fatal diseases, in which long-term survival is now achieved with cyclophosphamide (CYC) and prednisolone. As this standard treatment has shown considerable long-term morbidity and mortality and as more sensitive diagnostic procedures allow the earlier diagnosis of these diseases, nowadays, stage adapted treatment regimens that reduce the exposure to CYC are required. There is consensus that at present CYC remains the drug of choice in patients with generalized vasculitis for the induction of a remission period comprising 3-6 months. Whether pulse CYC is to be preferred over daily oral CYC is currently assessed in a RCT. There are efforts to further minimize the cumulative CYC dose for remission induction in elderly people, because the mortality is highest, and by adding monoclonal anti-B-cell antibodies. Adding Etanercept to the conventional induction regimen has not proven beneficial in a US RCT. For maintenance of remission a switch from CYC to azathioprine has proven to be safe. Methotrexate for this indication has been found to be comparable to azathioprine in one trial, but was associated with more relapses than leflunomide in another. Mycophenolate mofetil is currently studied with 48 months follow-up time. For induction of remission in patients without renal insufficiency and vital organ failure methotrexate at 0.3 mg/kg/week can replace CYC in patients with moderately extended disease and without pronounced granulomatous changes in the respiratory tract. Myfortic will be assessed for a similar indication in the future. Currently, long-term follow-up of the EUVAS patients is also sought. PMID- 16370165 TI - Autologous stem cell transplantation of treatment-resistant systemic vasculitis- a single center experience and review of the literature. AB - AIMS: Autologous peripheral blood stem cell transplantation (autoPBSCT) is increasingly and successfully applied to patients with treatment-resistant autoimmune diseases, mainly multiple sclerosis and systemic sclerosis, but also juvenile idiopathic arthritis and systemic lupus erythematosus. We intended to analyze the effects of autoPBSCT in patients with treatment-resistant systemic vasculitis by analyzing the outcome of 4 patients from our own hospital, and comparing them to cases reported in the literature. METHODS: 4 patients with treatment-resistant vasculitis (Wegener granulomatosis, Churg Strauss syndrome, Takayasu arteritis and relapsing polychondritis) received an autologous PBSCT. Stem cell mobilization was performed with cyclophosphamide (CY) and G-CSF, stem cells were purged by positively selecting CD34+ stem cells over a CliniMacs device, and the conditioning was performed with high dose CY and anti-thymocyte globulin (ATG). RESULTS: AutoPBSCT was well tolerated in all 4 patients. The patient with WG achieved complete remission although cANCA persisted, the other patients are in good partial remissions and respond to maintenance treatments which had been ineffective before PBSCT (CSA, azathioprin). Glucocorticosteroids (GC) could be reduced to a maximum of 10 mg in all patients. Shortly after the procedure, reactivation of viruses from the herpes family occurred in 3 of the patients and had to be treated. In the data base, 25 patients transplanted for severe systemic vasculitis are registered, in the literature, 6 additional vasculitis patients remitting after autoPBSCT are reported. CONCLUSIONS: Autologous PBSCT is feasible and effective in severe, treatment-resistant forms of systemic vasculitis. Data are sparse, further prospective studies are needed. These should also aim at evaluating more optimal regimens for conditioning and purging during PBSCT, as in most of the vasculitis patients reported until now, mostly good partial remissions, but less complete remissions were achieved. PMID- 16370166 TI - Coexistence of idiopathic membranous glomerulonephritis and idiopathic focal segmental glomerulosclerosis. PMID- 16370167 TI - Mycotic aneurysm in hemodialysis. PMID- 16370168 TI - [Testicular germ-cell tumour: diagnosis and treatment]. AB - Among germ cell tumours, seminomas hold a particular status related to their radio-sensitivity. Although radiotherapy remains the best treatment for Localized tumours of stage 1, in some cases, surveillance or chemotherapy may presently be considered as alternative therapies. Due to Long-term radiotherapy-related adverse effects, in particular the risk of second non-germ malignancies or cardiac morbidity, both dose and irradiation field are reduced in case of lymphatic retroperitoneal extension. Chemotherapy is the preferential treatment in more advanced stages, either with retroperitoneal bulky disease or with metastatic extension. Its efficacy allows Limiting surgical indications on residual masses, relying partly on the follow-up data of positron emission transaxial tomography assessment. PMID- 16370169 TI - [Imaging in the management of urinary tract infections]. AB - This review analyzes the role of imaging in the diagnosis, follow-up and management of urinary tract infections. Uncomplicated infection of the urinary tract is frequent and usually resolves rapidly with treatment; in such cases imaging is unnecessary except to exclude dilatation or the existence of an obstacle. Progression towards complex infection often occurs in patients with predisposing factors. Imaging helps in evaluating the extent of the disease; it has a role in the selection of therapy and interventional procedures. PMID- 16370170 TI - [Ganglion curage in prostate cancer]. AB - In clinically Localized prostate cancers, the interest of pelvic Lymphadenectomy is debated. Nevertheless, this intervention provides important information on disease prognosis (number of positive lymph nodes, tumoural volume, and extracapsular perforation of the affected ganglions); information that previously no other technique could provide. However, no consensus exists concerning patients who should benefit from pelvic Lymphadenectomy and on the extent of this intervention. For most surgeons, decision making regarding ganglion curage is based on nomograms. According to these nomograms, patients with a level of prostate specific antigen (PSA) <10 ng/mL and a Gleason score <7 have a very low risk for ganglionic metastases; this is the reason why the benefit of pelvic Lymphadenectomy remains controversial. Besides, most of these nomograms are based upon the results of standard Lymphadenectomy (iliac vein and obturator fossa) with, subsequently, a related risk of imprecision. In addition, potential therapeutic benefit may be expected from extended ganglion curage, despite the fact that this is not clearly documented yet, due to the benign course of the disease. In other tumoural diseases (stomach cancer, breast cancer, colorectal cancer, blade cancer), on the contrary, survival and stage identification depend on the number of removed ganglions, thus on the extent of Lymphadenectomy. PMID- 16370171 TI - [Laparoscopic pelvic lymphadenectomy: role of the surgeon assistant]. AB - Laparoscopy has emerged as a valid alternative to open surgery. Surgery assistance is a full part within the learning of Laparoscopy. Even more than in open surgery, efficient assistance facilitates the surgeon's work and helps shortening the intervention duration. Basic principles are detailed, such as the number and the position of trocars, or how to adequately manipulate optic and camera or suction devices. Specific gesture of the assistant during a pelvic Laparoscopic Lymphadenectomy is presented step by step, with the procedure undertaken by intra- or extra-peritoneal approach. PMID- 16370172 TI - [Prostate cancer: the essential from the congress of the American Urological Association (AUA) 2005]. AB - In 2005, the annual meeting of the American Urological Association (AUA) took place in San Antonio (Texas). Prostate cancer was one of the main topics. More than 400 worldwide presentations were dedicated to this field. The aim of this work is to present prostate cancer highlights of the 2005 AUA annual meeting, and in particular the main subjects of news and innovations that are supposed to modify urological clinical practice. Basic research studies were excluded from this work. What is related to Localized prostate cancer is first presented, followed by studies on locally advanced and metastasis disease. A third part summarizing the may 24 prostate cancer plenary sessions was added. PMID- 16370173 TI - Insight into traditional Chinese medicine obtained from an October 2004 visit to China organized by the People to People Ambassador Program. PMID- 16370174 TI - Increased dissolution rate and bioavailability through comicronization with microcrystalline cellulose. AB - Micronization is a commonly used enabling technology to improve the bioavailability of compounds where absorption is dissolution rate limited. However, decreasing particle size often results in increased Van der Waals' interactions and electrostatic attraction between particles. This causes agglomeration of particles, thereby compromising the increase in surface area gained by micronization. Comicronization with excipients has been reported to offer significant advantages over neat micronization. The present work describes the comicronization of a model compound CI-1040 at a high drug load that shows an increase in the dissolution rate and bioavailability in male Wistar rats. Physicochemical characterization of the comicronized and neat micronized material is presented to help explain the in-vitro and in-vivo data. PMID- 16370175 TI - The influence of magnesium stearate on the Hiestand Tableting Indices and other related mechanical properties of maltodextrins. AB - Magnesium stearate is widely used as a lubricant in pharmaceutical dosage forms. A film of magnesium stearate, or other lubricant, around individual granules is generally considered to cause the softening of tablets. Maltodextrins were chosen as model excipients since maltodextrins possessing a series of molecular weights showed systematically changing consolidation mechanisms. As maltodextrin molecular weight increases, the plasticity of the material increases. The Hiestand Tableting Indices and other related mechanical properties were used to study the effect of magnesium stearate addition on the mechanical properties and consolidation mechanisms of single component maltodextrins. Magnesium stearate was shown to lower the tensile strengths of maltodextrin compacts. Additionally, magnesium stearate addition lowered both the Hiestand Bonding Index and the Hiestand Brittle Fracture Index. The lower values of the Hiestand Brittle Fracture Index indicate that magnesium stearate promotes greater plastic behavior in maltodextrin compacts. PMID- 16370176 TI - Characteristics of rofecoxib-polyethylene glycol 4000 solid dispersions and tablets based on solid dispersions. AB - The aim of this work was to report the properties of rofecoxib-PEG 4000 solid dispersions and tablets prepared using rofecoxib solid dispersions. Rofecoxib is a poorly water soluble nonsteroidal anti-inflammatory drug with a poor dissolution profile. This work investigated the possibility of developing rofecoxib tablets, allowing fast, reproducible, and complete rofecoxib dissolution, by using rofecoxib solid dispersion in polyethylene glycol (PEG) 4000. Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to characterize the solid state of solid dispersions. The effect of PEG 4000 concentration on the dissolution rate of rofecoxib from its solid dispersions was investigated. The dissolution rate of rofecoxib from its solid dispersions increased with an increasing amount of PEG 4000. The extent of dissolution rate enhancement was estimated by calculating the mean dissolution time (MDT) values. The MDT of rofecoxib decreased significantly after preparing its solid dispersions with PEG 4000. The FTIR spectroscopic studies showed the stability of rofecoxib and absence of well-defined rofecoxib-PEG 4000 interaction. The DSC and XRD studies indicated the amorphous state of rofecoxib in solid dispersions of rofecoxib with PEG 4000. SEM pictures showed the formation of effective solid dispersions of rofecoxib with PEG 4000 since well defined change in the surface nature of rofecoxib and solid dispersions were observed. Solid dispersions formulation with highest drug dissolution rate (rofecoxib: PEG 4000 1:10 ratio) was used for the preparation of solid dispersion based rofecoxib tablets by the direct compression method. Solid dispersion-based rofecoxib tablets obtained by direct compression, with a hardness of 8.1 Kp exhibited rapid drug dissolution and produced quick anti-inflammatory activity when compared to conventional tablets containing pure rofecoxib at the same drug dosage. This indicated that the improved dissolution rate and quick anti inflammatory activity of rofecoxib can be obtained from its solid dispersion based oral tablets. PMID- 16370177 TI - Solid lipid microparticles (SLM) containing juniper oil as anti-acne topical carriers: preliminary studies. AB - Solid lipid microparticles (SLM) were used as carriers of juniper oil and proposed for the topical treatment of acne vulgare. The formulations were obtained by the o/w emulsification method. Compritol and Precirol were employed as lipidic materials. Emulsions containing 1.5% (w/w) of lipophilic phase (lipid and oil) and two different lipid to oil ratios (1:1 and 2:1) were prepared. Blank particles were also prepared, as a comparison. The SLM were characterized in terms of encapsulation efficiency, size, and morphology. The particle size stability in aqueous dispersions was monitored over one month. Evaporation of volatile compounds of oil from microparticles by weight loss was investigated. The qualitative composition of Juniper oil before and after the encapsulation process was determined by gas chromatography (GC) and gas chromatography/mass spectrum (GC/MS) analyses. The antimicrobial activity of the oil encapsulated into the lipid microparticles against P. acnes was studied as contact time assay and compared to the activity of the oil not encapsulated. The emulsification method here described was a good technique for the encapsulation of essential oils. Percentage yields of production and encapsulation efficiencies were higher for Compritol preparations than for these prepared using Precirol. All preparations were characterized by similar particle size distributions (dvs about 3-4 microm) regardless of lipid type and lipid to oil ratios. Microscopy observations showed that the microparticles in aqueous dispersions had almost spherical shape, independently from their composition. The scanning electron microscopy (SEM) analyses showed that when the particles were dried, they had an irregular shape and a rough surface. The SLM dispersions based on Compritol revealed particle size stability over the investigated period of 30 days. In contrast, an increase of the mean dimensions in the preparations containing Precirol was observed. A low loss of volatile oil compounds owing to evaporation from dry particles was found in all preparations. This indicated that the microparticles were able to substantially maintain the oil loaded inside their lipidic structure, reducing its volatility. Some modifications of composition were found in the oil encapsulated in SLM with respect to the juniper oil raw material, but these modifications did not decrease the antibacterial activity of the oil. The SLM here described are promising carriers for the development of anti-acne topical formulations containing Juniper oil. PMID- 16370178 TI - Behavior of moisture gain and equilibrium moisture contents (EMC) of various drug substances and correlation with compendial information on hygroscopicity and loss on drying. AB - The behavior of moisture gain and equilibrium moisture content (EMC) was determined for 30 drug substances at relative humidities (RH) ranging from 11% to 93%. Based on the results, the drugs were categorized into different classes, following the classification system proposed by Callahan et al. About 23 fell under Class-I (nonhygroscopic), four under Class-II (slightly hygroscopic), and three under Class-III (moderately hygroscopic). Most of the Class-III drugs converted from solid to liquid state at > 75% RH, and the moisture increase was more than 40% above 90% RH. However, the moisture increase was comparatively much smaller at humidities < 40%-50%. Hence, no drug could be categorized in Class-IV (very hygroscopic), where gain of moisture is generally higher even at lower humidities. The results were correlated to the statements given in the compendia on hygroscopicity, and also the values of loss on drying (LOD). The study suggests that there is a need to rationalize the pharmacopoeial information. PMID- 16370179 TI - Drug-excipient compatibility testing using a high-throughput approach and statistical design. AB - The aim of our research was to develop a miniaturized high throughput drug excipient compatibility test. Experiments were planned and evaluated using statistical experimental design. Binary mixtures of a drug, acetylsalicylic acid, or fluoxetine hydrochloride, and of excipients commonly used in solid dosage forms were prepared at a ratio of approximately 1:100 in 96-well microtiter plates. Samples were exposed to different temperature (40 degrees C/ 50 degrees C) and humidity (10%/75%) for different time (1 week/4 weeks), and chemical drug degradation was analyzed using a fast gradient high pressure liquid chromatography (HPLC). Categorical statistical design was applied to identify the effects and interactions of time, temperature, humidity, and excipient on drug degradation. Acetylsalicylic acid was least stable in the presence of magnesium stearate, dibasic calcium phosphate, or sodium starch glycolate. Fluoxetine hydrochloride exhibited a marked degradation only with lactose. Factor interaction plots revealed that the relative humidity had the strongest effect on the drug excipient blends tested. In conclusion, the developed technique enables fast drug-excipient compatibility testing and identification of interactions. Since only 0.1 mg of drug is needed per data point, fast rational preselection of the pharmaceutical additives can be performed early in solid dosage form development. PMID- 16370180 TI - Purification of pharmaceutical excipients with supercritical fluid extraction. AB - Supercritical fluid extraction (SFE), with carbon dioxide as the solvent, was tested for its ability to remove common reactive impurities from several pharmaceutical excipient powders including starch, microcrystalline cellulose (MCC), hydroxypropylcellulose (HPC), polyethylene oxide (PEO), and polyvinylpyrrolidone (PVP). Extraction of the small molecule impurities, formic acid and formaldehyde, was conducted using SFE methods under conditions that did not result in visible physical changes to polymeric excipient powders. It could be shown that spiked, largely surface-bound, impurities could be removed effectively; however, SFE could only remove embedded impurities in the excipient particles after significant exposure times due to slow diffusion of the impurities to the particle surfaces. Attempts at hydrogen peroxide extraction were hindered by its low solubility in CO2, thereby effectively precluding SFE for removal of hydrogen peroxide from excipients. This work suggests that SFE will only be commercially useful for removal of low molecular weight impurities in polymeric excipients when migration of the impurities to the particle surfaces is sufficiently rapid for extraction to be completed in a reasonable time frame. PMID- 16370181 TI - Drug-drug interaction studies on first-line anti-tuberculosis drugs. AB - The purpose of this study was to carry out drug-drug compatibility studies on pure first line anti-tuberculosis drugs, viz., rifampicin (R), isoniazid (H), pyrazinamide (Z), and ethambutol hydrochloride (E). Various possible binary, ternary, and quaternary combinations of the four drugs were subjected to accelerated stability test conditions of 40 degrees C and 75% relative humidity (RH) for 3 months. For comparison, parallel studies were also conducted on single drugs. Changes were looked for in the samples drawn after 15, 30, 60, and 90 days of storage. Analyses for R, H, and Z were carried out using a validated HPLC method. The E was analyzed by liquid chromatography-tandem mass spectrometry (LC MS/MS), as it does not absorb in ultraviolet (UV). All single pure drugs were relatively stable and showed only 3%-5% degradation under accelerated conditions for 3 months. However, significant interactions were observed in case of the drug mixtures. In particular, ternary and quaternary drug combinations containing R and H along with Z and/or E were very unstable, showing 90%-95% and 70%-75% loss of R and H, respectively. In all these cases, isonicotinyl hydrazone (HYD) of 3 formylrifamycin and H was found to be the major degradation product. In case of RE and RZE mixtures, where H was absent, 3-formylrifamycin was instead the key degradation product. Another unidentified peak was observed in the mixture containing RZE. Apart from these chemical changes, considerable physical changes were also observed in pure E and the mixtures containing E, viz., RE, ZE, RHE, RZE, and RHZE. In addition, significant physical changes associated with noteworthy loss of H and E were also observed in mixtures containing HE and HZE. The present study thus amply shows that the four primary anti-tuberculosis drugs, when present together, interact with each other in a multiple and complex manner. PMID- 16370183 TI - ["Only he who is willing to give, should also get"]. PMID- 16370182 TI - A potent and selective tacrine analog--biomembrane permeation and physicochemical characterization. AB - Cholinesterase inhibitors have been used for years in treatment of Alzheimer's disease (AD). Tacrine is the first acetylcholinesterase inhibitor approved for treating AD by the regulatory agencies around the world. Unfortunately, a number of studies have shown tacrine to be associated with some severe side effects, including hepatotoxicity. These adverse effects may be attributed to its poor selectivity for acetylcholinesterase and have thus necessitated the research and development of more selective cholinesterase inhibitors with a greater specificity and higher potency. The heptylene-linked bis-tacrine analog (bis-THA) of Tacrine is a second-generation inhibitor of acetylcholinesterase, which has a potency that is 1000 times more potent than Tacrine in inhibition of the rat brain acetylcholinesterase and 10,000 times more selective for acetylcholinesterase over butyrylcholinesterase. A series of investigations have thus been initiated to characterize the physicochemical properties (e.g., pKa, partition coefficient, and stability) of this bis-THA analog as compared to its parent molecule (Tacrine). For AD treatment, the cholinesterase inhibitors need to be taken daily for long periods of time. Use of controlled-release dosage forms to deliver drugs for chronic administration, by taking advantage of their rate-controlling drug delivery features, has gained increasing popularity in recent years. On the other hand, the nasal route, which has been used to deliver drugs for achieving a direct delivery to the brain (via the olfactory pathway), could offer the benefits of brain targeting to the delivery of Tacrine and bis THA. To investigate this feasibility, the permeation of Tacrine and bis-THA across the nasal mucosa was evaluated (in comparison with other absorptive mucosae). Studies of their permeation kinetics across the various absorptive mucosae, which were freshly excised from the domestic pig, indicated that the nasal mucosa could present a viable pathway for the systemic delivery of bis-THA. Delipidization studies suggested that the lipophilic components in the absorptive mucosae could play an important role in the permeation of bis-THA. The bis-THA has a pKa of approximately 8 and its partition coefficient showed a sigmoidal pattern with solution pHs. It was found to be relatively stable at acidic pHs but subjected to a base-catalyzed degradation at the alkaline pHs (> or = 8) and at higher temperatures (> or = 50 degrees C). PMID- 16370184 TI - [Is organ donation still questionable? (interview by Dr. med. Jochen Aumiller)]. PMID- 16370185 TI - [Prescriptions for insomnia]. PMID- 16370186 TI - [Psychogenic paralysis--somatogenic psychosis]. PMID- 16370187 TI - [Long-term study shows increased risk. Cancer from cellular phone lines?]. PMID- 16370188 TI - [Death with dignity? Assisted death after diagnostic error]. PMID- 16370189 TI - [Human connection instead of assisted suicide]. PMID- 16370190 TI - [What do you prescribe for climacteric symptoms? (interview by Friederike Klein)]. PMID- 16370191 TI - [This is how Finland wants to conquer infarction and fatal infarction]. PMID- 16370192 TI - [Diagnosis and treatment of deep leg vein thrombosis in the physician's office. Awareness of a danger helps to avoid it]. PMID- 16370193 TI - [Diagnosing deep vein thrombosis in the leg]. AB - As a consequence of changes in lifestyle, an ageing population sometimes overdoing things in terms of sports activities, long-haul travel and an increasing incidence of malignomas, deep vein thrombosis (DVT) is increasingly being seen in ambulatory patients. The incidence of DVT is some 0.1-0.3%, and increases with age. With ultrasound and phlebography the exclusion of a thrombosis is almost always possible in an ambulatory setting. Diagnostic algorithms involving the standardized determination of clinical probability (CP) and measurement of D-dimers can help to reduce diagnostic problems in the doctor's office. A low CP in combination with a negative D-dimer excludes a DVT with virtual certainty. PMID- 16370194 TI - [Treatment options in deep vein thrombosis]. AB - In the event of a deep vein thrombosis, the major concern is to prevent the subsequent development of an embolism or a recurrence. The therapeutic time gap to the development of the full effect of the vitamin K antagonists is bridged in the initial acute treatment phase with low molecular weight heparin (LMWH) or fondaparinux. In patients in whom vitamin K antagonists are contraindicated, or in pregnancy, secondary prophylaxis is alternatively continued with dose-adjusted LMWH. Compression therapy serves merely to prevent a post-thrombotic syndrome. PMID- 16370195 TI - [Current evaluation of the fluoroquinolones]. AB - Originally, the fluoroquinolones were classified into the groups I-IV on the basis of their clinical field of application, pharmacokinetics, antibacterial spectrum, and bioavailability. In the present publication an evaluation taking account in particular of aspects of clinical application and dosage has been undertaken based on the 1998 classification by a group of PEG experts. PMID- 16370196 TI - [Prevention of metabolic syndrome and type 2 diabetes]. AB - Type 2 diabetes does not represent inescapable fate, but is an avoidable disease, when the main prophylactic approach targets the central problem of insulin resistance. This means daily physical exercise, the reduction of the glycemic load in the diet and--provided there are no contraindications--certainly a daily glass of wine. PMID- 16370197 TI - [Coalition contract examined. A posse with a tragic end for physicians]. PMID- 16370198 TI - [New epidemiologic data show: conventional NSAIDs need individual benefit-risk evaluation]. PMID- 16370199 TI - [Study results of routine clinical practice. Do rheumatism patients get substandard care?]. PMID- 16370200 TI - [Hardly any hypoglycemias, constant weight--and still cost effective]. PMID- 16370201 TI - [After 40 years of age, normal weight is the exception]. PMID- 16370202 TI - [General practice under white sails]. PMID- 16370203 TI - [What should be considered in acute diarrhea?]. AB - This article provides an overview of possible causes, differential diagnosis, diagnostics and therapy of acute diarrhea. PMID- 16370204 TI - [Red circles over the whole body]. PMID- 16370205 TI - [Total body irradiation: techniques and main indications]. AB - The radiotherapy department at Salah Azaiz institute had started, in March 2000, a new sophisticated technique of irradiation consisting in total body irradiation (TBI). TBI is used in many preparative regimens before bone marrow transplantation in the treatment of haematological malignancies. TBI aims to destroy immunocompetent tissues in order to avoid graft rejection and to eradicate residual tumor cells. In this article, we review different TBI techniques and its main indications. We also describe the acute and late effects of TBI. PMID- 16370206 TI - [Chronic aortic regurgitation with left ventricular dysfonction]. AB - We studied fifty one patients (40 men and 11 women) under going valve replacement from 1990 to 2002 for aortic regurgitation and had left ventricular dysfunction. 45% patients were in class III or IV of New York Heart Association (NYHA). All patients were investigated by echocardiography (left ventricular ejection fraction [LVEF] < 50%) - Peroperative mortality was 5.8% due to myocardial failure. 81.4% of survivors were followed duering a mean period of 24 months (rangis from 3 to 67 months) after valve replacement. PMID- 16370207 TI - [Variation of biochemical parameters of the first morning urine during month of Ramadan]. AB - The purpose of this study is to evaluate the effect of fluid and diet restriction in fasting on biochemical factors of stone formation. Our study concernes 90 patients divided in three groups: healthy fasting patient (GI), healthy non fasting patient (G2) and non fasting patient with calcium lithiasis (G3). The promotors (oxalate, calcium, uric acid, phosphates) and inhibitors (citrate, magnesium) are statistically significant between G1, G2 and G3, G2. Supersaturation of urine with oxalate, uric acid and brushite are the same for (G1) and (G3) and higher than (G2). Crystalluria is more important in lithiasis subjects compared with healthy non fasting patients (58% vs 11,4%). Oxalate monohydrate (Whewellite) and uric crystal don't exist in the healthy non fasting people but reached 4% and 12% respectively in the lithiasis patient. The crystalluria profil is the same in the heathy fasting patients and calcium lithiasis patients. However healthy patients have equilibria between promotors and inhibitors of crystal formation which minimize the risk of crystalluria and subsequent stone formation. PMID- 16370208 TI - [Bacteriological profile of bacteremias in badly burnt patients]. AB - The purpose of this work is to study the bacteriological profiles of germ responsible for bacteriemas / septicemias in badly burnt patients hospitalized in burns utis, in order to guide better the antibiotherapy of first intention. During the period of survey going from January 1st, 2001 to December 31st,2002 71 patients had one or several episodes of bateremia. The average burnt cutaneous surface was 48.7%. All collected bacteriemas were contacted in hospial. Empirical antbiotheray was effective in 30 patients. The hospital stay for this group was long: 45 to 86 days. Acinetobacter baumannii was the most incriminated germ(20.3%), followed klebsiella pneumoniea (12.7%). The frequency of resistance was variable according to the considered species. PMID- 16370209 TI - [Children Helicobacter pylori infection: antibiogram results]. AB - Helicobacter pylori infection is frequent in children. The eradication rate is variable, it is influenced by resistance of the bacteria to the antibiotics used. The aim of this study was to determine the resistance rate of the most commonly used antibiotics among helicobacter pylori (Hp) strains isolated in infected children. PATIENTS AND METHODS: 60 children (26 males, 34 females), with mean age of 8 years 8 months (3 years and 2 month (2.5-13.5 years) underwent an oesogastroscopy. Forty six of them had helicobacter infection assessed by the presence of Hp on the gastric biopsy and or positivity of urease test. The infected children were treated with three medication; Omeprazole. Clarythromycin, and Amoxicillin. The eradication of Hp was confirmed in 13 patients by the absence of Hp on the gastric biopsy and/or negativation of the urease test; RESULTS: Culture was positive in 13 infected children susceptibility to anti microbial drugs was done in ten children. Nine children had primary metronidazole resistance, three had both clarythromycin and Metronidazole resistance, three had Clarythromycin primary resistance. No case of Amoxicillin primary resistance was detected. Children eradication was achieved in four children among 13 children. This study shows the difficulty of Hp culture. It confirms the absence of Hp resistance to Amoxicilline. the high level of Hp resistance to Metronidazole and the mild level of Clarythromycin resistance. The authors stress the importance of Antibiotic susceptibility assay in helicobacter pylori infections. PMID- 16370210 TI - [Abruptio placentae]. AB - Abruptio Placentae is one of the most typical obstetrical emergencies. MATERIALS AND METHODS: We present a retrospective study about 45 cases of placental abruption. The diagnosis was confirmed in all cases by both clinical and anatomopathological findings. OBJECTIVES: Study the epidemiological profile of the patient, the circumstances of diagnosis, the modes of delivery and the fetal morbidity. RESULTS: The prevalence of placental abruption was about 0.34%. Patients were primiparous in 40% of the cases. This problem occurred during the third trimester in 97.8% of the cases. Elevated blood pressure was found in 44.4% of the cases. The incomplete form represented 77.8% of cases. Blood clotting disorders were detected in 8 patients. Fetal death rate was 37.77%. CONCLUSION: This problem remains unpredictable and sudden. Both maternal morbidity and fetal mortality rates remain high. PMID- 16370211 TI - [The results of anterior cruciate ligament reconstrution by bone--patellar tendon -bone technique. A series of 40 cases]. AB - We reviewed retrospectively 40 patients treated surgically for rupture of anterior cruciate ligament. The technique used was arthroscopy bone patellar tendon reconstruction. The mean age of patients was 27 years. The operative delay was 27.2 months in average. Functional and anatomic result was evaluated with a mean follow up of 24 months. 80% of patients were classified IKDC A or B. Sports activity was resumed after 6.6 months in average. Results depented meniscus lesions and operative delay. Reconstruction of anterior cruciate ligament with bone patellar tendon bone is a reliable technique. Operative delay should be as short as possible to prevent meniscus lesion and aggravation of laxity. PMID- 16370212 TI - [Pronostic factors in squamous cell carcinoma of the vulva. About 35 cases]. AB - OBJECTIVE: To evaluate the pronostic factors of the vulvar cancer. STUDY DESIGN: This is a retrospective study, of 35 cases of squamous cell carcinoma of the vulva diagnosed and treated at Farhat Hached Hospital (Sousse) between January 1991 and December 2002. The study of the pronostic factors is based on analysis of the following parameters obtained after a period varyinf from 6 months to 5 years: clinical data, pre-therapeutique and therapeutic evaluation, pathologic data, outcome and survival rate. RESULTS: The diagnostic was often late with a delay for consultation superior to 6 months in 60% of cases. 80% of the patient wete in stages III and IV of FIGO with an average tumorous size of 3.8 cm and palpable lymph node in 74.2% of the cases. 22 patient underwent vulvectomy with bilateral inguinal node dissection. Adjuvant therapy was carried out in 8 cases. The rate of relapses was 22.7%. The global survival at 5 years of the operated patients was of 40%. The main prognostic factors were: the size tumorale, the degrees of stromal invasion and the lymph node invasion. CONCLUSION: Early detection fo vulvar concer by regular gynecologic examinations and a biopsy of all suspect lesions, allows an early treatment consequently a better prognosis. PMID- 16370213 TI - [Ocular complications in peritoneal haemodialysis]. AB - In this work, we consider the results of a study carried out in our unit about 65 dialysed chronic renal failure patients. The work is based on the study of medical records and the results of ophtalmoscopic examination and phosphocalcic balance evaluation. Ocular manifestations and their relation with initial nephropathy, dialysis and phosphocalcic disturbances are described. The following alterations were observed: - corneo conjunctival deposits (60%), - cataract (26%), - hypertensive retinopathy in 21(68%) cases among 31 (48%) who suffered from hypertension, - hypertensive choroidopathy in one case, - vascular thrombosis in one case, - diabetic retinopathy in all patients (7) with diabetes. PMID- 16370214 TI - [Dermatofibrosarcoma protuberans. About 18 cases]. AB - Dermatofibrosarcoma protuberans (DFSP) is a rare mesenchyma skin tumor. It is characterized by a slow growth with a high rate of recurrence but limited potential for metastasis. We report the result of a retrospective study of 18 cases of DFSP collected over 16 years in the dermatology department of La Rabta Hospital. The mean age was 32 years with a sex ratio of 1.57. The site of predilection was the trunk (77.5%). Histological features were typical in all cases with positivity for CD34 in 14 cases. Treatment was surgical in 13 cases. DFSP is a tumor of intermediate malignancy, which can have aggressive course. This tumor is best treated with surgery in which large excision are necessary to reduce the risk of recurrence. PMID- 16370215 TI - [Embolization of head and neck hypervascular lesions. A retrospective study of 5 cases]. AB - Embolisation of head and neck hypervascular tumours and arterioveinous malformations (AVMs) is now a well-established therapeutic procedure. The purpose of this study was to analyse the technique and to evaluate the safety and value of preoperative embolization of hypervascular lesions. We retrospectively reviewed the records of 5 patients, agged between 14 and 47 years, with hypervascular tumours and AVMs in the head and neck. Lesions were distributed as follow: nasal angiofibroma (n = I), nasal angioleiomyoma (n = I), nasopharyngeal angiofibroma (n = I), aneurysmal bone cyst in the posterior element of the second cervical vertebra (n = I) and AVM of the inferior lip (n = I). Angiography and embolization were carried out at the same time. Complete exclusion of lesions was obtained without any complications. Complementary surgical resection was performed with not notable blood loss. PMID- 16370216 TI - [Morgagni hernia in elderly patients: a study about two cases]. AB - Two cases of retrocostoxiphoid hernia are reported. Both patients were adults. The hernia was strangled in one case and symptomatic in the second case. The diagnosis was confirmed by radiology. Reduction of the visceral hernia and closure of the orifice was carried out after laparotomy. A review of literature and of clinicopathological features are given in the present article. Histogenesis and differential diagnosis are also discussed. PMID- 16370217 TI - [Successful use of levosimendan in a patient with cardiogenic shock complicating acute myocardial infarction]. AB - Levosimendan is a new inodilator that improves cardiac contractility by sensitizing troponin C to calcium. This drug has proved to be effective in treating advanced congestive heart failure but has not been evaluated in cardiogenic shock. We present the case of a 54-year-old male patient treated successfully with levosimendan for cardiogenic shock following acute myocardial infarction. Treatment with dobulamine. revascularisation and itra-aortic balloon conterpulsation had first failed to improve his hemodynamic variables. Levosimendan induced a steady decline of increased pulmonary capillary wedge pressure, followed by an increase in cardiac index and mixed venous oxygen saturation. Left ventricular ejection fraction improved from 25% to 47%. Infusion of levosimendan can be used in cardiogenic shock without side effects and to improve hemodynamics and left ventricular function. PMID- 16370218 TI - [Septic oligoarthritis as a complication of rhumatoid arthritis--a case report]. AB - We present the case a 53-year-old patient followed-up since 1999, for erosive AR treated with methotrexate and glucocorticoids. In April 2000, he had an arthritis of the right knee. The identification of an enterobacter in blood culture, and synovial biopsy results permitted the diagnosis of septic arthritis. After 23 days of antibioterapy treatment, the patient had an arthritis of the left knee. The infectious origin was confirmed by synovial biopsy. The course was better after adaptation of the antibiotics. Septic arthritis is then a serious complication of AR. It requires a fast and multidisciplinary management. It can be threatenig in fragile and immunocompromised patients. The functional prognosis is especially compromised in polyarticular septic arthritis. PMID- 16370219 TI - [Orbital eosinophil granuloma. Case report and review of the literature]. AB - Eosinophyl granuloma is a rare tumor effecting mainly children, involving the vault more frequently than the skull base. Treatment is mainly surgical, adjuvant therapies can be used only in special cases. The report is about he casde of a 8 year-old girl o presented with a right frontal mass and a non-significant exophtalmia; Neuroradilogical investigations showed a non-painful frontolateral mass with bone erosion in regard. Surgery revealed an invasive tumor for the temporal bone and muscles. Histological, epidemiological and radiological features are discussed in the study. PMID- 16370220 TI - [Uterin choriocarcinoma revealed by pulmonary metastasis. A case report]. AB - Uterine choriocarcinoma is a rare trophoblastic tumor, characterized by its metastatic potentiality and its sensitivity to chemotherapy. Pulmonary metastases are frequent, but they rarely reveal the disease. We report the case of a 30-year old patient with uterine choriocarcinoma. The disease occurred after a normal delivery and was revealed by a pulmonary metastasis. Through of this case and a review of literature, we remind of the diagnostic and prognostic particularities of uterine choriocarcinoma. PMID- 16370221 TI - Calcium and exercise affect the growing skeleton. AB - Adequate dietary calcium and bone-stimulating exercise during growth are known to affect skeletal development, but the combined effects of dietary calcium and osteogenic exercise have received scant attention. Animal research has showed a compensatory effect of impact loading on calcium-deprived bones, while various human studies have suggested compensatory, additive, or possibly synergistic effects in certain skeletal locations. Current evidence suggests that the best strategy for strong bones by the end of childhood may be either high-impact exercise with a moderate or greater calcium intake or a combination of moderate impact exercise and adequate calcium during growth. PMID- 16370222 TI - Components of olive oil and chemoprevention of colorectal cancer. AB - Olive oil contains a vast range of substances such as monounsaturated free fatty acids (e.g., oleic acid), hydrocarbon squalene, tocopherols, aroma components, and phenolic compounds. Higher consumption of olive oil is considered the hallmark of the traditional Mediterranean diet, which has been associated with low incidence and prevalence of cancer, including colorectal cancer. The anticancer properties of olive oil have been attributed to its high levels of monounsaturated fatty acids, squalene, tocopherols, and phenolic compounds. Nevertheless, there is a growing interest in studying the role of olive oil phenolics in carcinogenesis. This review aims to provide an overview of the relationship between olive oil phenolics and colorectal cancer, in particular summarizing the epidemiologic, in vitro, cellular, and animal studies on antioxidant and anticarcinogenic effects of olive oil phenolics. PMID- 16370223 TI - Disruption of methyl group metabolism by ethanol. AB - Recent studies have focused on establishing a link between the pathogenesis of ethanol and the disruption of metabolic pathways in the liver. Ethanol alters hepatic methionine metabolism, leading to perturbation of S-adenosylmethionine dependent transmethylation. Therefore, the supply of metabolically related nutrients such as folate may play a role in the hepatotoxic effects of ethanol. PMID- 16370224 TI - Do soy isoflavones cause endometrial hyperplasia? AB - For many years, hormone replacement therapy (HRT) was considered the gold standard for the symptomatic treatment of menopause. Clinical trials have found that HRT reduces the symptoms of hot flashes and sweating, while also decreasing vaginal dryness and urinary tract infections. HRT has also been shown to be protective against colon cancer (37%) and hip fractures (34%). However, recent findings from the Women's Health Initiative (WHI) have revealed that long-term HRT may actually lead to an increase in heart disease (29%), breast cancer (26%) and other adverse events, such as stroke (41%). Consequently, many women in the United States and abroad are actively looking for alternative treatments for menopause, including botanical dietary supplements. PMID- 16370225 TI - 5,10-Methylenetetrahydrofolate reductase polymorphisms and pharmacogenetics: a new role of single nucleotide polymorphisms in the folate metabolic pathway in human health and disease. AB - Knowledge about the role of folate, a water-soluble B vitamin, and single nucleotide polymorphisms (SNPs) in the folate metabolic pathway in human health and disease has been rapidly expanding. Recently, functionally significant SNPs in 5,10-methylenetetrahydrofolate reductase (MTHFR), a critical enzyme for intracellular folate homeostasis and metabolism, have been identified and characterized. An emerging body of in vitro and clinical evidence suggests that these MTHFR SNPs may be an important pharmacogenetic determinant of predicting response to and toxicity of methotrexate and 5-fluorouracil-based cancer and anti inflammatory treatments because of their well-defined and highly relevant biochemical effects on intracellular folate composition and one-carbon transfer reactions. PMID- 16370226 TI - Helping young people paint the canvas of life. PMID- 16370227 TI - Hand hygiene, an essential process in the OR. PMID- 16370230 TI - What about the floor? PMID- 16370231 TI - Substance abuse among nurses--defining the issue. AB - The prevalence of substance abuse in the nurse population is believed to parallel that in the general population (i.e. approximately 10%). Nurses with substance abuse problems need help. They are in danger of harming patients, the facility's reputation, the nursing profession, and themselves. The consequences of not reporting concerns can be far worse than those of reporting the issue. Part one of this two-part series discusses how society views addiction and the nursing profession, signs and symptoms of substance abuse, reasons nurses should report an impaired colleague, the code of silence that exists among nurses, and board of nursing jurisdiction. PMID- 16370232 TI - Pulmonary thromboendarterectomy for thromboembolic pulmonary hypertension. AB - Pulmonary artery thromboendarterectomy (PTE) is a surgical procedure that offers the only cure for chronic thromboembolic pulmonary hypertension (TPH), a progressive form of secondary pulmonary hypertension. Current estimates indicate that 1% to 5% of patients who survive a pulmonary embolus will develop chronic TPH with progressively worsening pulmonary hypertension and right-sided heart failure. This bilateral procedure is performed through a sternotomy incision using cardiopulmonary bypass, profound hypothermia, and intermittent circulatory arrest with antegrade cerebral perfusion. As an alternative to lung transplantation, PTE offers patients with chronic TPH an improved quality of life. PMID- 16370233 TI - Orthopedic personnel roles in the OR for computer assisted total knee arthroplasty. AB - The methods and problems associated with traditional total knee arthroplasty surgery and the development and advantages of computer-assisted surgery (CAS) are discussed in this article. In the next decade, CAS products have the potential to transform as many as 25% of all orthopedic surgical procedures from techniques that largely depend on line of sight and traditional instruments to techniques that use integrated surgical instruments and computer technology. This technology could revolutionize orthopedic surgery, the lives of patients, and the work of perioperative staff members during the next decade. PMID- 16370234 TI - Developments in the RN first assistant role during the Korean War. AB - During times of war, demand for health care providers has stimulated increased employment of nurses and broadened the dimensions of the scope of nursing practice in the surgical arena. This article examines how the work of army nurses during the Korean War helped develop the role of the RN first assistant. PMID- 16370235 TI - Congress leaves its mark on students and their mentors. PMID- 16370236 TI - Perioperative initiatives for medication safety. PMID- 16370237 TI - pH-sensitive behavior of two-component hydrogels composed of N,O-carboxymethyl chitosan and alginate. AB - A two-component pH-sensitive hydrogel system composed of a water-soluble chitosan derivative (N,O-carboxymethyl chitosan, NOCC) and alginate cross-linked by genipin, glutaraldehyde or Ca2+ was investigated. Preparation and structures of these hydrogels and their swelling characteristics and release profiles of a model protein drug (bovine serum albumin, BSA) in simulated gastrointestinal media are reported. At pH 1.2, the swelling ratios of the hydrogels cross-linked by distinct methods were limited. Of note is that the lowest swelling ratios of test hydrogels were found at pH 4.0. At pH 7.4, the carboxylic acid groups on test hydrogels became progressively ionized and led to a significant swelling. There was barely any BSA released from the glutaraldehyde-cross-linked hydrogel throughout the entire course of the study. The amounts of BSA released at pH 1.2 from the genipin- and Ca(2+)-cross-linked hydrogels were relatively low (approx. 20%). At pH 4.0, there was still significant BSA release from the Ca(2+)-cross linked hydrogel, while the cumulative BSA released from the genipin-cross-linked hydrogel was limited due to its shrinking behavior. At pH 7.4, the amount of BSA released from the genipin- and Ca(2+)-cross-linked hydrogels increased significantly (approx. 80%) because the swelling of both test hydrogels increased considerably. The aforementioned results indicated that the swelling behaviors and drug-release profiles of these test hydrogels are significantly different due to their distinct cross-linking structures. PMID- 16370238 TI - Preparation of a hyaluronic acid hydrogel through polyion complex formation using cationic polylactide-based microspheres as a biodegradable cross-linking agent. AB - A novel hyaluronic acid (HA)-based hydrogel was prepared through polyion complex (PIC) formation between cationic polylactide (PLA)-based microspheres (MS+) and hyaluronic acid (HA-) as an anionic polyelectrolyte. The MS+ and HA formed a biodegradable PIC hydrogel (HA-/MS+) when mixed in aqueous media. The swelling behavior and mechanical properties of the PIC hydrogel could be controlled by changing the charge ratio between HA- and MS+. In addition, the HA-/MS+ PIC hydrogel resulted in a lower inflammatory response compared with a collagen hydrogel in vivo. PMID- 16370239 TI - Novel biodegradable films and scaffolds of chitosan blended with poly(3 hydroxybutyrate). AB - In order to develop a novel biomaterial, films of chitosan blended with poly(3 hydroxybutyrate) (PHB) were prepared by an emulsion blending technique and their properties were characterized. Scanning electron microscopy (SEM) showed that PHB microspheres were formed and were entrapped in chitosan matrices, which made the film surface rough. With increasing PHB content, the roughness of the film surface increased, while the swelling capability of the films decreased. In a wet state, the blended films exhibited a lower elastic modulus, a higher elongation at-break and a higher tensile strength compared with chitosan films. Cell-culture experiments revealed that the blended films had better cytocompatibility than chitosan films. To explore the potential application of the blended material in tissue engineering, the porous blended scaffolds were fabricated and their pore morphology was observed by SEM. The results revealed that not only pore structure but also pore wall morphology of the blended scaffolds could be controlled by selecting the parameters of the fabrication process. These advantageous properties indicate that the blended chitosan/PHB material is promising for tissue engineering applications. PMID- 16370240 TI - Properties of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) films modified with polyvinylpyrrolidone and behavior of MC3T3-E1 osteoblasts cultured on the blended films. AB - A series of composite films of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) modified with polyvinylpyrrolidone (PVP) was prepared by varying the ratio of constituents, and their properties and cytocompatibility were evaluated. The hydrophilicity of the blended materials surfaces increased and the amounts of fibronectin and laminin adsorbed on the materials surface increased remarkably compared with PHBHHx. FT-IR spectra of the blended films showed a new band, implying that a surface physical interpenetrating network structure had formed. Scanning electron microscopy showed that there were dense pits and holes on the blended films surface. For the films of PHBHHx with 20 wt% and 40 wt% PVP, MTT assay indicated that PVP enhanced cell adhesion and proliferation, but that the effects were impaired by excessive PVP. The results suggested that proper addition of PVP increased the cytocompatibility of PHBHHx because the material surface had increased hydrophilicity and presented an appropriate morphology. PMID- 16370241 TI - In vivo tumor targeting of ODN-PEG-folic acid/PEI polyelectrolyte complex micelles. AB - A tumor-targeting antisense oligodeoxynucleotide (ODN) delivery system based on polyelectrolyte complex (PEC) micelles is demonstrated. ODN-PEG-folic acid (ODN PEG-FA) was synthesized using a heterofunctional PEG linker. The PEC micelles for the targeted ODN delivery to tumor cells were produced by ionic interactions between the ODN-PEG-FA and polyethylenimine (PEI). The in vivo targeting properties of the PEC micelles were assessed using a mouse tumor model. The size of ODN-PEG-FA/PEI PEC micelles was 92.3 nm with a relatively narrow distribution. Cellular uptake of the ODN-PEG-FA/PEI PEC micelles by folic acid receptor over expressing cells (KB) was greatly enhanced compared to that of ODN-PEG/PEI PEC micelles. When the ODN-PEG-FA/PEI PEC micelles were systemically administered to the mice bearing KB cell xenograft tumor, ODN was accumulated to the solid tumor in a target specific manner. This study suggests that the PEC micelles with a receptor-recognizable targeting ligand on the surface have potential for passive and active targeted delivery of ODN drugs to cancer cells. PMID- 16370242 TI - A thermo-sensitive poly(organophosphazene) hydrogel used as an extracellular matrix for artificial pancreas. AB - A poly(organophosphazene) bearing alpha-amino-omega-methyl-poly(ethylene glycol) (AMPEG) and hydrophobic L-isoleucine ethyl ester (IleOEt) side groups has been synthesized. This material exhibited 4 phase transitions in an aqueous solution on gradually increasing the temperature, i.e., a transparent sol, a transparent gel, an opaque gel and a turbid sol. A 10 wt% buffered solution of the polymer was employed to entrap islets of Langerhans in an artificial pancreas. Rat islets entrapped in the gel showed prolonged insulin secretion in response to basal (5.5 mM) glucose concentration compared to free rat islets and islets entrapped in other types of polymer gels. Over a 28-day culture period, the rat islets in the poly(organophosphazene) hydrogel maintained higher cell viability and insulin production versus rat islets in different hydrogels and free islets. This thermo sensitive injectable, biodegradable matrix can be used with several cell types, including nerve cells, to promote nerve regeneration. PMID- 16370243 TI - Sol/gel transition of chitosan solutions. AB - This work studies the occurrence of sol/gel transition and the gel rheology for chitosan solution under various conditions. Experiments were conducted in an oscillatory shear apparatus with small amplitude, using a Rheometrics SR-5 rheometer, with Couette and parallel plate geometries. The experimental results demonstrate that the sol/gel transition concentration and the elastic modulus (G') for CS gel decrease as the pH value and the molecular weight (Mw) increase. However, the sol/gel transition concentration and G' became independent of Mw when Mw exceeded a threshold. The higher ionization constant, Kp, is responsible for the higher sol/gel transition concentration in a formic acid solution than in an acetic acid solution with equivalent molar concentration. The elastic modulus G' of a CS gel increases with temperature, which relationship differs from that for many polysaccharides, and can be understood through classical rubber elastic theory. Finally, a gel whose concentration was barely above the sol/gel point exhibited aging, and its G' and G" declined rather than increase with time, accompanied by a reversal from the sol/gel state back to the sol state. This is an uncommon aging behavior for a polysaccharide and a detailed explanation is provided. PMID- 16370244 TI - Preparation of poly(ethylene glycol)-introduced cationized gelatin as a non-viral gene carrier. AB - The objective of this study was to prepare cationized gelatins grafted with poly(ethylene glycol) (PEG) (PEG-cationized gelatin) and evaluate the in vivo efficiency as a non-viral gene carrier. Cationized gelatin was prepared by chemical introduction of ethylenediamine to the carboxyl groups of gelatin. PEG with one terminal of active ester group was coupled to the amino groups of cationized gelatin to prepare PEG-cationized gelatins. Electrophoretic experiments revealed that the PEG-cationized gelatin with low PEGylation degrees was complexed with a plasmid DNA of luciferase, in remarked contrast to that with high PEGylation degrees. When the plasmid DNA complexed with the cationized gelatin or PEG-cationized gelatin was mixed with deoxyribonuclease I (DNase I) in solution to evaluate the resistance to enzymatic degradation, stronger protection effect of the PEG-cationized gelatin was observed than that of the cationized gelatin. The complex of plasmid DNA and PEG-cationized gelatin had an apparent molecular size of about 300 nm and almost zero surface charge. These findings indicate that the PEG-cationized gelatin-plasmid DNA complex has a nano-order structure where the plasmid DNA is covered with PEG molecules. When the PEG cationized gelatin-plasmid DNA complex was intramuscularly injected, the level of gene expression was significantly increased compared with the injection of plasmid DNA solution. It is concluded that the PEG-cationized gelatin was a promising non-viral gene carrier to enhance gene expression in vivo. PMID- 16370245 TI - [Epidermal growth factor receptor (EGFR): therapeutic target in the treatment of lung adenocarcinoma]. AB - Revolution in biotechnology made possible to identify those gene errors, which via their encoded proteins (mostly kinase enzymes) are key players in tumor development, growth and progression, and could be considered as molecular targets in tumor diagnosis and therapy. Activity of EGFR (epidermal growth factor receptor), an outstanding representative of the regulatory cell surface receptors, can be inhibited by drugs proved for clinical use. In the past year many groups observed that those lung adenocarcinoma cells, which contain activating mutation in the tyrosine kinase domain of EGFR show remarkable sensitivity to anti-EGFR compounds. The basis of the effective therapy is the identification of the mutations. The clinical advantage of EGFR is an example from the coming age of tumor chemotherapy, when the presence of molecular targets will guide the therapeutic choice. PMID- 16370246 TI - [Use of self-expanding stents in benign and malignant esophageal disease]. AB - Of patients with malignant esophageal tumors, more than the half has incurable disease at the time of diagnosis. In such cases, quality of life and ability to swallow should be improved or restored by palliating the process e.g. by inserting esophageal stents. In the recent years, use of covered self-expandable metal stents (SEMS) has led to an increased success rate in the palliative treatment of--mainly malignant--strictures of the esophagus. High occurrence of complications (perforation, bleeding) associated with the use of rigid plastic esophageal stents necessitated the development of flexible stents. First generation self-expandable stents were made of special metal alloys, such as nickel-titanium. In the recent years, self-expandable plastic stents are available, too, offering new alternatives in the palliative treatment of esophageal diseases. In our present review article, we discuss the most important aspects about the use of self-expanding esophageal stents, based on recent clinical observations and data. PMID- 16370247 TI - [Review of effectiveness of chemotherapy with gemcitabine in ovarian cancer]. AB - Ovarian cancers are known to have the highest mortality rate among the gynaecological tumours. Despite the effectiveness of the paclitaxel-carboplatin combination that is usually recommended nowadays the disease does recur, many times causing finally the death of the patient. This phenomenon explains the continual efforts to find further new and potent treatment forms in this disease. From the late 1990 years on the nucleoside analogue gemcitabine has been used against ovarian cancer. Its administration in Hungary was approved in 2005. Publications report on 16.2 +/- 5.6% and 53.9 +/- 20.5% complete response on average with mono- and polychemotherapy using gemcitabine in ovarian cancer. Some chemotherapeutic combinations with gemcitabine for the first line treatment of ovarian cancers are also known and they were found to be superior to the multi line treatments (81.6 +/- 17.9% complete remission on average with the former). These treatments are usually accompanied by haematological toxicity. Severe G3-4 neutropenia was observed in 45.1 +/- 22.6%, and thrombocytopenia in 24.9 +/- 23.1% of the patients. The first publications on this treatment form are promising, however, a final policy in this matter can only be assumed after finishing the evaluation of the comprehensive, randomised studies (GOG-0182 ICON5) in progress. PMID- 16370248 TI - [A population-based case-control study of broncho-vaxom use during pregnancy]. AB - OBJECTIVE: To study human teratogenic potential of oral broncho-vaxom treatments during pregnancy. AIM: Pair analysis of cases with congenital abnormalities and matched population controls without congenital abnormalities. PATIENTS AND METHOD: The large population-based data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996. PARTICIPANTS: Of 38.151 pregnant women who had babies without any defects (control group), and 22.865 pregnant women who had newborn infants or fetuses with congenital abnormalities (case group). RESULTS: In the control group 58 (0,15%), while in the case group 22 (0,10%) pregnant women were treated with broncho-vaxom (OR with 95% CI: 0.6, 0.4-1.1). The case-control pair analysis concerning the use of broncho-vaxom during the second-third months of pregnancy did not show any human teratogenic potential in the different groups of congenital abnormalities. The mean gestational age was longer in control infants born to mother with broncho-vaxom treatment during pregnancy compared with the figure of control infants without this treatment (40 +/- 1.3 vs. 39.4 +/- 2.1, t = 2.28, p = 0.02). CONCLUSION: Teratogenic risk for congenital abnormalities was not detectable in the children born to women treated with broncho-vaxom during pregnancy. The possible gestational age promoter effect of broncho-vaxom needs further studies. PMID- 16370249 TI - [Commencement address at the end of the 236th academic year]. PMID- 16370250 TI - [Istvan Benedek (1915-1996) the culture historian, physician writer]. PMID- 16370251 TI - [Remarks about "The medical grand round"]. PMID- 16370252 TI - Perceived barriers to teaching for critical thinking by BSN nursing faculty. AB - The ability to think critically is considered an essential skill of nursing graduates and competent nursing practice. Yet, the literature reports that teachers are having difficulty teaching for critical thinking and that critical thinking is lacking in new nursing graduates. This research study sought to identify barriers to the implementation of critical thinking teaching strategies by nursing faculty currently teaching in generic baccalaureate programs in Tennessee. Surveys were mailed to 262 nursing faculty; 194 were returned, and 175 were usable. Students' attitudes and expectations represented the single greatest barrier to the implementation of critical thinking teaching strategies, followed by time constraints and the perceived need to teach for content coverage. Recommendations to support and encourage faculty to teach for critical thinking are outlined. PMID- 16370253 TI - An evidence-based curriculum to prepare students for global nursing practice. AB - This article describes a curriculum designed to prepare nurses for global public health practice. Designed to be adapted to meet the needs of either undergraduate or graduate students, the curriculum uses the Internet to provide the knowledge and skills needed by nurses to effectively practice in areas around the globe. This course offering integrates the disciplines of nursing and public health with state-of-the-art technology to teach nurses how to identify the health care needs of populations, prioritize national and international responses, and design health care delivery services to meet these needs. PMID- 16370254 TI - Uncovering racial bias in nursing fundamentals textbooks. AB - This article describes research that sought to identify and critique selected content areas from three nursing fundamentals textbooks for the presence or absence of racial bias embedded in the portrayal of African Americans. The analyzed content areas were the history of nursing, cultural content, and physical assessment/hygiene parameters. A researcher-developed guide was used for data collection and analysis of textual language, illustrations, linguistics, and references. A thematic analysis resulted in I I themes reflecting the portrayal of African Americans in these sampled textbooks. An interpretive analysis with a lens of Sadker and Sadker's categories of bias, along with other literary and theoretical contexts, were used to explore for the presence or absence of racial bias. Recommendations for nursing education are provided. PMID- 16370255 TI - Lessons learned from developing, implementing, and evaluating a model of community-driven nursing. AB - The Helene Fuld Health Trust, HSBC Bank USA,Trustee has made generous contributions to the development of community-based nursing education. As a recipient of funds, faculty at the University ofWisconsin-Madison School of Nursing developed, implemented, and evaluated a pilot curriculum for undergraduate nursing students. The first phase of the project changed the landscape of community-based nursing education through faculty development. Unlike traditional approaches to faculty development, a resident expert model of faculty development based on dialogue was used. The second phase changed the landscape of teaching and learning community-based care through curricular and instructional revisions. Noteworthy changes were the creation of a curriculum responsive to both individual student and community needs, and recruitment of a cadre of teachers to enhance the cost-effectiveness of clinical education. The third phase changed the landscape of community-based nursing education through evaluation research. Several strategies were used to assess the project's usefulness, including interpretive phenomenological study of interview data and review of conventional outcome measures. Each phase of the project contributed to a new vision of community-based nursing education, a vision in which nursing as service is taught and learned. PMID- 16370256 TI - Development of an academic consortium for nurse-managed primary care. AB - This article describes the development of an academic consortium comprised of four universities and their schools and colleges of nursing. The initial efforts at collaboration arose from the need to obtain funding and address policy issues related to advanced practice issues at the state level. The developmental stages of the Michigan Academic Consortium are described within a framework developed by Bailey and Koney (I). PMID- 16370257 TI - A Renaissance in nursing education thriving in a new era. PMID- 16370258 TI - Desensitization to strong vestibular stimuli improves tolerance to simulated aircraft motion. AB - INTRODUCTION: The extent to which tolerance acquired in one motion environment can be transferred to another has not been fully elucidated. This study compares three provocative motion stimuli and whether tolerance acquired from cross coupled motion and torso rotation can be transferred to simulated aircraft motion. METHOD: There were 18 healthy subjects (9 men and 9 women), 18-44 yr of age, who were divided into 3 groups. Each group was exposed to the following desensitization procedure in a pseudo-random repeated measures design. In condition D1, subjects were exposed to 4 consecutive days of cross-coupling and on the 5th d they were exposed to simulated aircraft motion. In D2, subjects were exposed to 4 consecutive days of torso rotation and on the 5th day they were exposed to the simulated aircraft motion as in D1. In D3, subjects were exposed to 5 consecutive days of simulated aircraft motion. Based on Graybiel's diagnostic criteria, severity of motion sickness on the 1st and 5th day in D1, D2, and D3 were compared. RESULTS: Cross-coupled motion was the most provocative stimulus followed by torso rotation and simulated aircraft motion. Within each motion stimulus, there was evidence of desensitization (p < 0.001) within 4 d. There were no significant differences in the severity of motion sickness observed on the 5th day regardless of the desensitization stimuli. CONCLUSION: Our results suggest that desensitization to one provocative motion could be transferred to a less severe motion stimulus. This result could be used in the refinement of desensitization programs for aircrew. PMID- 16370259 TI - Cardiovascular neuroregulation during acute exposure to 40, 70, and 100% oxygen at sea level. AB - BACKGROUND: Humans encounter increased partial pressures of inspired oxygen in some kinds of diving as well as during use of hyperoxic mixtures to shorten decompression times and hyperbaric oxygen therapy for decompression sickness or other clinical conditions. Although it is known that hyperoxia may affect cardiovascular regulation, such effects are generally obscured by stress and the diving reflex. In this study, we evaluated cardiovascular neuroregulation for various levels of hyperoxia in a laboratory setting. METHODS: There were 10 healthy adults who were exposed to 21, 40, 70, and 100% oxygen administered via mask as a series of stepwise increases. Subjects breathed at a fixed respiratory rate of 15 breaths x min(-1) while their heart rate (HR) and blood pressure (BP) were measured continuously over 5-min intervals. RESULTS: HR decreased with increasing fraction of inspired oxygen (FIO2) (21%: 65 +/- 9, 40%: 63 +/- 9, 70%: 61 +/- 8, 100%: 60 +/- 8 bpm) and the high-frequency power of HR variability (an index of cardiac parasympathetic activity) increased as FIO2 rose (21%: 773 +/- 565, 40%: 880 +/- 590, 70%: 966 +/- 681, 100%: 1114 +/- 715 ms2); both changes were significant at the 70% and 100% oxygen levels. The low-frequency power of systolic BP variability (an index of vasomotor sympathetic activity) did not change. Low- and high-frequency transfer function gains (indices of arterial cardiac baroreflex function) increased with FIO2. CONCLUSION: Parasympathetic activity and arterial-cardiac baroreflex function increased with hyperoxia in a dose-dependent manner. This increase may help reduce the likelihood of arrhythmias during diving. PMID- 16370260 TI - Motion sickness symptoms in a ship motion simulator: effects of inside, outside, and no view. AB - INTRODUCTION: Vehicle motion characteristics differ between air, road, and sea environments, both vestibularly and visually. Effects of vision on motion sickness have been studied before, though less systematically in a naval setting. It is hypothesized that appropriate visual information on self-motion is beneficial in a naval setting and that task performance is likely reduced as sickness increases. METHODS: Using a within-subjects design, 24 subjects were exposed to 30 min of motion in a ship's bridge motion simulator with 3 visual conditions: an Earth-fixed outside view; an inside view that moved with the subjects; and a blindfolded condition. Subjective sickness symptoms and severity were rated repeatedly before, during, and after motion exposure. During the motion, subjects performed a mental task. RESULTS: Though not excessive, sickness was highest in the inside viewing condition, intermediate in the outside viewing condition, and least in the blindfolded condition. The blindfolded condition was equally as bad as the inside viewing condition during the first 5-10 min of motion exposure. The overall temporal increase of sickness during motion was about equal to the decrease during recovery. No effect of sickness on task performance was observed. DISCUSSION: Most sickness in a naval setting is observed when the visual environment moves with the subjects, as has been reported in other environments, such as cars. Only mild sickness, caused by moderate motions, was provoked in this study and was alleviated by the performance task. A non-linear brain mechanism integrating visual and vestibular information may explain why the least sickness was observed when subjects were blindfolded. PMID- 16370261 TI - Resistance Index of Frostbite as a predictor of cold injury in arctic operations. AB - INTRODUCTION: Cold-induced vasodilation (CIVD) is mentioned as a mechanism that may prevent the occurrence of local cold injuries. The magnitude of the CIVD reaction differs considerably between subjects and there were some indications that those subjects with a fast CIVD reaction with high amplitude had a reduced risk for cold injuries. The purpose of this investigation was to determine the magnitude of the finger CIVD reaction in subjects prior to operation in cold areas and to relate these scores to the occurrence of cold injuries. METHODS: In order to evaluate the magnitude of the CIVD response, 206 subjects immersed their left middle finger in ice water for 30 min. The Resistance Index for Frostbite (RIF) according to Yoshimura was determined on the basis of the finger skin temperature response. This index ranges from 3 (high risk) to 9 (low risk) depending on the response time and response magnitude. Later, most of the screened subjects deployed as part of a group of 1080 marines for winter operations in Norway. RESULTS: The Caucasian subjects in this study had higher RIF scores than the non-Caucasians (7.0 +/- 1.6 vs. 6.1 +/- 2.1). The mean RIF was relatively high as compared with a reference group of Japanese male soldiers (6.9 +/- 1.7 vs. 5.7 +/- 1.7). Unexpectedly, smokers had a higher RIF score than non-smokers did. The RIF score was inversely related to pain during the test. There were 54 marines who suffered cold injuries during training in Norway. Of those, 11 were in the measured group of 206 marines. These subjects had a RIF of 5.2 +/- 1.6, as compared with 7.0 +/- 1.6 for the remaining subjects, which was significantly different. CONCLUSIONS: The RIF shows considerable differences between subjects. The RIF, determined in a simple lab test, may be related to the risk for cold injuries during operations in the field. PMID- 16370262 TI - Muscle characteristics in career breath-hold divers: effect of water temperature. AB - INTRODUCTION: In a previous study we reported that Korean female breath-hold divers (BHD) with life-long experience of diving in cold water (10-12 degrees C in winter and 25-27 degrees C in summer) had reduced muscle fiber size and increased capillary density. The hypothesis tested in the present study was whether prolonged habitual diving at a moderate water temperature (MWT, 29-30 degrees C all year round) similarly caused a reduction in muscle fiber size. METHODS: The subjects were 14 Indonesian BHDs with long experience of diving at MWT, and a control group of 10 age-matched non-diving Indonesian men (CON), selected from the same tribe among which the BHDs lived. Muscle samples obtained from the middle portion of the vastus lateralis muscle were analyzed for muscle morphology by histochemical analysis and the levels of vascular endothelial growth factor (VEGF) protein by Western blotting. RESULTS: Muscle fiber type composition was identical in both groups, and no difference in cross-sectional area (CSA), VEGF protein, or capillarity between the BHD and the CON was observed. CONCLUSION: The present study demonstrated that prolonged habitual breath-hold diving at MWT does not cause any alteration in muscle fiber composition, fiber size, or capillarity. PMID- 16370263 TI - Hearing loss in military aviation and other trades: investigation of prevalence and risk factors. AB - INTRODUCTION: Continuous increase in the cost of claims for noise-induced hearing loss in Canadian Forces personnel prompted a review of hearing conservation practices. Investigations comprised retrospective analyses of hearing test results and a prospective survey of risk factors for hearing loss. METHOD: There were 1057 individuals working in 107 air, land, and sea trades who contributed their current and first hearing test results. Subjects completed a 64-item questionnaire relating to demographics, occupational and non-occupational noise exposure history, training in and use of hearing protection, and non-noise risk factors, including head injury, ear disease, medications, and solvent exposure. Subject recruitment, hearing assessment, and questionnaire distribution were carried out by military personnel. Apparatus and protocols for hearing testing conformed to current clinical practice. RESULTS: Prevalence of moderate to severe hearing loss progressed with age, with hearing thresholds in those over 45 yr ranging broadly from normal to over 70 dB HL. Unprotected exposure to noise and solvents appeared to be the likely determinates of adverse outcome. Head injury, ear disease, and the use of ototoxic medications had minimal impact. The data for individuals in 17 selected trades in the 3 environments modeled that of the total sample. DISCUSSION: Responses to the questionnaire suggested that training on noise hazard and selection and use of hearing protection were inadequate. Hearing protection was reported to be incompatible with other gear, uncomfortable, and an impediment to communication. Approaches for upgrading hearing conservation strategies, including the creation of end-user focus groups, were considered. PMID- 16370264 TI - Toxicological findings from 1587 civil aviation accident pilot fatalities, 1999 2003. AB - INTRODUCTION: The prevalence of drug and ethanol use in aviation is monitored by the Federal Aviation Administration (FAA). Under such monitoring, toxicological studies for the 1989-1993 and 1994-1998 periods indicated lower percentages of the presence of controlled substances (illegal drugs) than that of prescription and nonprescription (over-the-counter) drugs in aviation accident pilot fatalities. In continuation, a toxicological assessment was made for an additional period of 5 yr. METHODS: Biosamples from aviation accident pilot fatalities submitted to the FAA Civil Aerospace Medical Institute (CAMI) are analyzed, and those findings are stored in a database. This database was examined for the 1999-2003 period for the presence of controlled substances (Schedules I V), prescription/nonprescription drugs, and ethanol in the fatalities. RESULTS: Out of 1629 fatal aviation accidents from which CAMI received biosamples, pilots were fatally injured in 1587 accidents. Drugs and/or ethanol were found in 830 (52%) of the 1587 fatalities. Controlled substances from Schedules I-II (SI-II) and Schedules III-V (SIII-V) were detected in 113 and 42 pilots, respectively. Prescription drugs were present in 315 pilots, nonprescription drugs in 259, and ethanol in 101. SI-II substances were detected in 5 of 122 first-class medical certificate-holding airline transport pilots. In addition to the controlled substances, many of the prescription/nonprescription drugs found in the fatalities have the potential for impairing performance. CONCLUSIONS: Findings from this study were consistent with those of two previous toxicological studies and support the FAA's programs aimed at reducing the usage of performance impairing substances. PMID- 16370265 TI - Aerobatic flight effects on baroreflex sensitivity and sympathovagal balance in experienced pilots. AB - BACKGROUND: Aerobatic flights subject pilots to accelerations and, therefore, to heavy physical workloads. OBJECTIVE: Our aim was to document changes in spontaneous baroreflex sensitivity and disturbances of sympathovagal balance after exposure to "push-pull" accelerations. METHODS: During 30-min flights, five aerobatic pilots performed five series of descending spirals: first, 30 s under negative (-3 Gz max), and then 30 s under positive (+4 Gz max) G loading, climbing between each series to regain altitude. A stand-test was performed before (T0), immediately postflight (PF), 1 h (PF1), and 2 h after (PF2) the flight. A Finapres apparatus recorded heart rate (HR) and BP during the stand tests. RESULTS: Resting HR was higher at PF than T0 in supine (11.2 +/- 5.3%, p < 0.01) and standing (11.0 +/- 4.9%; p < 0.05) positions. Sequence analysis of spontaneous baroflex sensitivity (BRS) and spectral analysis of HR variability showed that: a) supine spontaneous BRS did not differ between preflight and postflight, while parasympathetic modulation of HR variability tended to increase; and b) supine spontaneous BRS was higher at PF1 than PF (PF: 0.011 +/- 0.0014 ms x mmHg(-1), PF1: 0.015 +/- 0.0012 ms x mmHg(-1); p < 0.05) and parasympathetic modulation of HR variability (high frequency component) was higher at PF2 than PF (PF: 0.014 +/- 0.007, PF2: 0.039 +/- 0.009; p < 0.001). CONCLUSIONS: These findings may reflect a change in the sympathovagal balance during the second hour of recovery from repeated push-pull maneuvers. PMID- 16370266 TI - Aeronautical decision making: instructor-pilot evaluation of five mnemonic methods. AB - INTRODUCTION: The ability to make good decisions is a critical component of pilot proficiency. Results of recent research suggest that aeronautical decision making (ADM) can be improved by training. The purposes of this research are to identify the best ADM mnemonic-based methods for training military pilots in decision making in tactical environments. METHODS: There were 60 instructor pilots in the Republic of China Air Force Academy who evaluated the suitability of 5 different ADM methods including SHOR (Stimuli, Hypotheses, Options, Response); PASS (Problem identification, Acquire information, Survey strategy, Select strategy); FOR-DEC (Facts, Options, Risks & Benefits, Decision, Execution, Check); SOAR (Situation, Options, Act, Repeat); and DESIDE (Detect, Estimate, Set safety objectives, Identify, Do, Evaluate). Each was evaluated for six different types of decisions: go/no go; recognition-primed; response selection; resource management; non-diagnostic procedural; and creative problem-solving. RESULTS: The instructor-pilots regarded the SHOR method as the best for time-limited and critical, urgent situations, while they preferred DESIDE for knowledge-based decisions that needed more comprehensive consideration and were less time limited. DISCUSSION: To optimize training effectiveness, it is necessary to provide instruction based around both the SHOR and DESIDE decision-making mnemonic methods to provide comprehensive instruction for coping with all situations in the demanding military aviation environment. PMID- 16370267 TI - Tumor necrosis factor alpha inhibitors and methotrexate: implications for deployed personnel. AB - Rheumatic diseases such as rheumatoid and psoriatic arthritis are often diagnosed in younger age groups and are, therefore, likely to be encountered in active-duty military populations. These patients are increasingly being treated with disease modifying anti-rheumatic drugs (DMARDs) and biologic agents such as tumor necrosis factor alpha (TNF-alpha) inhibitors. While these classes of drugs have revolutionized the treatment of rheumatic diseases, they are also associated with serious potential adverse effects. At present, there are no published guidelines for the routine monitoring of laboratory parameters in patients receiving anti TNF therapy. Currently, no official consensus among military physicians exists regarding duty and geographic limitations for patients receiving these types of therapy. Major adverse effects of these agents are reviewed in this article. A survey of U.S. Air Force, Army, and Navy rheumatologists was performed. The results of laboratory monitoring and operational deployment recommendations are reported. The majority of U.S. military rheumatologists do not recommend deployment of patients while taking methotrexate or TNF-alpha inhibitors. PMID- 16370268 TI - Head-up tilt test for recurrent syncope in pilots. AB - Syncope is defined as a sudden temporary loss of consciousness and postural tone that is associated with spontaneous recovery. Vasovagal or neurocardiogenic syncope is a common and usually benign cause of syncope. The mechanism may be cardioinhibitory, vasodepressor, or both. Diagnosis is usually made by a typical patient history with a definite trigger. Although vasovagal syncope is considered a benign condition, its occurrence in an aviator is worrisome, especially if recurrent and without a definite trigger. The head-up tilt test (HUTT) is used as a vasovagal syncope challenge test. A drop in BP and asystole during HUTT suggest a tendency to recurrent vasovagal syncope. We describe two military aviators with recurrent episodes of vasovagal syncope, one with definite triggers and one without. Both had positive HUTTs. The aviator with trigger-defined vasovagal syncope was disqualified from high-performance platforms due to his positive HUTT. The second case was disqualified from all platforms, irrespective of his HUTT result, because a definite trigger was not definable for all his syncopal episodes. PMID- 16370269 TI - Cellulitis in Grade III ankle sprain requiring treatment for antibiotic-resistant bacteria. AB - This case report describes a novel occurrence of a Grade III ankle sprain complicated by cellulitis. The patient ultimately required aspiration of an extra articular fluid collection and treatment with parenteral antibiotics due to worsening of the infection despite treatment with second-generation cephalosporins. This individual's infection was consistent with MRSA-cellulitis. Cellulitis, in the context of a preceding ankle sprain, has not been reported in the literature; the resultant edema from the injury may have served as the nidus for infection. MRSA comprises a significant proportion of soft-tissue infections in the ambulatory setting, and physicians should incorporate this trend into therapeutic strategies for their infected patients. PMID- 16370270 TI - The Aerochir: the first "flying hospital". AB - The concept of providing light and far-forward mobile surgical support teams to care for battle casualties is being increasingly adopted by most NATO nations as a result of military and political-economic considerations. Similar considerations led to the design of a mobile surgical hospital capable of flying a surgical team and all its equipment to a location of need as early as 1919. Unlike many of the early attempts to fuse medicine and aviation, the Nemirovsky Tilmant Aerochir was not a dead-end. It served as a proof-of-concept product which led to the development and adoption of the Breguet "Aerochir," which was used in French colonial wars of the 1920s, and eventually led to adoption of the concept of air-mobile far-forward surgical care by many of the world's militaries. This design from over 80 yr ago, along with its creators, should be remembered and honored by those of us involved in providing aeromedical support to battlefield casualties. PMID- 16370271 TI - Decompression sickness during simulated EVA. PMID- 16370272 TI - A new era. PMID- 16370273 TI - What is ISO? PMID- 16370274 TI - Congenital hydrocephalus. PMID- 16370275 TI - Time trends and demographics of deaths from congenital hydrocephalus in children in the United States: National Center for Health Statistics data, 1979 to 1998. AB - OBJECT: Congenital hydrocephalus has an estimated population incidence of 0.2 to 0.8/1000 live births. With improvements in techniques for cerebrospinal fluid shunting, treatment of hydrocephalus has become safe and routine, yet data describing mortality from congenital hydrocephalus or demonstrating improvements in mortality with the advent of modern treatment are scarce. The authors' analysis sought to rectify this situation. METHODS: The authors performed an electronic search of National Center for Health Statistics death certificate databases to identify deaths from 1979 to 1998 attributed to congenital hydrocephalus, spina bifida with hydrocephalus, and acquired hydrocephalus (both obstructive and communicating) in all children in the US aged 1 day to 20 years. Mortality rates were defined as deaths per 100,000 person-years and were analyzed for differences on the basis of age, race, sex, and year. The authors identified 10,406 deaths attributed to childhood hydrocephalus within the 20-year study period. The overall mortality rate was 0.71 per 100,00 person-years. Mortality rates were highest in infants, with 3979 deaths; they were similar between girls and boys. Compared with white infants, black infants had higher relative risk (RR) for death caused by congenital hydrocephalus (RR 1.46, p value < 0.0001) and acquired hydrocephalus (RR 2.58, p value < 0.0001) but not for that caused by hydrocephalus with spina bifida (RR 0.65, p value < 0.0001). From 1979 to 1998, the mortality rate due to congenital hydrocephalus declined 66.3%, from spina bifida with hydrocephalus it declined by 30.4%, and from acquired hydrocephalus it declined by 67.5%. CONCLUSIONS: Mortality rates from childhood hydrocephalus have declined in US children over the previous 20 years. Black race is associated with higher mortality rates in infants for both congenital and acquired hydrocephalus, whereas sex has no effect. PMID- 16370276 TI - The role of lumbar shunts in the management of slit ventricles: does the slit ventricle syndrome exist? AB - OBJECT: Slit-ventricle syndrome (SVS) is a confusing description of presentations in patients with chronic shunt-treated hydrocephalus. These patients are prone to acute deterioration with recurrent malfunction. The authors describe the clinical profile and management outcome of using lumboperitoneal (LP) shunts in this population of patients. METHODS: Thirty-three patients with slit ventricles and recurrent malfunctions were converted to LP shunts (mean age 12 years). The initial ventricular shunt was placed at a mean age of 16.5 months. Ten patients had failed endoscopic third ventriculostomies prior to placement of their LP shunt. At a previous presentation, in 11 patients suspected to have SVS following revision of the shunt, intracranial pressure normalized after insertion of a contralateral shunt, suggesting that their ventricles were isolated. The rate of infection and malfunction was compared before and after conversion to an LP shunt. Twenty-seven patients were successfully converted to LP shunts. Four of the 11 patients with isolated ventricles required ventricular shunts in addition to the LP shunt. During a mean follow-up period of 16.7 months, the malfunction rate per patient decreased from 4.81 for ventriculoperitoneal shunts, prior to conversion to 1.48 after conversion to LP shunts, a statistically significant reduction (p < 0.000). No significant difference was found in the rate of shunt infections (7.1% for VP shunts and 9.6% for LP shunts, p = 0.44). No patient presented with acute symptoms following malfunction of an LP shunt or suffered from a Chiari I malformation. CONCLUSIONS: Conversion to an LP shunt is a safe and effective procedure in patients prone to rapid decompensation and recurrent shunt malfunctions from small, slitlike ventricles. The term SVS is confusing. The condition is a manifestation of an unrecognized slitlike isolated ventricle and should be abandoned. PMID- 16370277 TI - Use of rapid-sequence magnetic resonance imaging for evaluation of hydrocephalus in children. AB - OBJECT: The authors examine the use of rapid-sequence magnetic resonance (rsMR) imaging to make the diagnosis of malfunctioning and/or infected shunts in patients with hydrocephalus. Computerized tomography (CT) scanning is usually used in this context because it rapidly acquires high-quality images, yet it exposes pediatric patients to particularly high levels of radiation. Standard MR imaging requires longer image acquisition time, is associated with movement artifact, and, in children, usually requires sedation. Standard MR imaging provides greater structural resolution, yet visualization of ventricular catheters is relatively poor. METHODS: The authors analyzed a series of 67 rsMR imaging examinations performed without sedation in pediatric patients with hydrocephalus whose mean age was 4 years at the time of the examination. The mean study duration was 22 minutes. Catheter visualization was good or excellent in more than 75% of studies reviewed, and image quality was good or excellent in more than 60% of studies reviewed. The authors analyzed cancer risk with a model used for atomic bomb survivors. Fifty percent of their patients with hydrocephalus had undergone more than four brain imaging studies (CT or MR imaging) in their lifetimes. For the many patients who had undergone more than 15 studies, the total estimated lifetime attributable cancer mortality risk was calculated to be at least 0.35%. CONCLUSIONS: Rapid-sequence MR imaging yields reliable visualization of the ventricular catheter and offers superior anatomical detail while limiting radiation exposure. The authors' protocol is rapid and each image is acquired separately; therefore, motion artifact is reduced and the need for sedation is eliminated. They recommend the use of rsMR imaging for nonemergent evaluation of pediatric hydrocephalus. PMID- 16370278 TI - Effect of antibiotic-impregnated shunt catheters in decreasing the incidence of shunt infection in the treatment of hydrocephalus. AB - OBJECT: The majority of shunt infections occur within 6 months of shunt placement and chiefly result from perioperative colonization of shunt components by skin flora. Antibiotic-impregnated shunt (AIS) systems have been designed to prevent such colonization. In this study, the authors evaluate the incidence of shunt infection after introduction of an AIS system in a population of children with hydrocephalus. METHODS: The authors retrospectively reviewed all pediatric patients who had undergone cerebrospinal fluid (CSF) shunt insertion at their institution over a 3-year period between April 2001 and March 2004. During the 18 months prior to October 2002, all CSF shunts included standard, nonimpregnated catheters. During the 18 months after October 2002, all CSF shunts included antibiotic-impregnated catheters. All patients were followed up for 6 months after shunt surgery, and all shunt-related complications, including shunt infection, were evaluated. The independent association of AIS catheter use with subsequent shunt infection was assessed via multivariate proportional hazards regression analysis. A total of 211 pediatric patients underwent 353 shunt placement procedures. In the 18 months prior to October 2002, 208 (59%) shunts were placed with nonimpregnated catheters; 145 (41%) shunts were placed with AIS catheters in the 18 months after October 2002. Of patients with nonimpregnated catheters, 25 (12%) experienced shunt infection, whereas only two patients (1.4%) with antibiotic-impregnated catheters experienced shunt infection within the 6 month follow-up period (p < 0.01). Adjusting for intercohort differences via multivariate analysis, AIS catheters were independently associated with a 2.4 fold decreased likelihood of shunt infection. CONCLUSIONS: The AIS catheter significantly reduced incidence of CSF shunt infection in children with hydrocephalus during the early postoperative period (< 6 months). The AIS system used is an effective instrument to prevent perioperative colonization of CSF shunt components. PMID- 16370279 TI - Intracranial volume in patients with nonsyndromal craniosynostosis. AB - OBJECT: In recent years, comparisons between intracranial volumes (ICVs) in patients with craniosynostosis and healthy patients have given variable results, leading to questions regarding the validity of the normal reference material and the comparability of the measurement techniques. In this study, ICVs in patients with nonsyndromal craniosynostosis without previous surgical intervention were compared with the ICVs of a normal population of European descent determined using the same method for each group. METHODS: Determination of ICV was based on measuring the area of intersection in each computerized tomography slice. For comparisons the ICV measurements for each patient were standardized with regard to age and sex by expressing them in terms of the standard deviation score. Only the group of boys with metopic synostosis had a tendency toward smaller ICV than did healthy boys (p = 0.04). Partitioning the male metopic data into age groups younger and older than 7 months of age revealed that the younger children had normal ICVs, whereas the older children had, on average, smaller ICVs (p = 0.02). Both the female sagittal synostosis and the male unilateral coronal synostosis groups had larger than normal ICVs, both with a probability value less than 0.001. CONCLUSIONS: No evidence was found that the ICVs of patients with nonsyndromal craniosynostosis are smaller than those of normal children, except for boys older than 7 months of age with metopic synostosis. This finding may have implications for the timing of surgical intervention for these patients. The indications are that interventions should be focused less on ICV and more on normalizing craniofacial shape and promoting normal development. PMID- 16370280 TI - Surgical implications of the thickened pituitary stalk accompanied by central diabetes insipidus. AB - OBJECT: The authors discuss the indications for and timing of a diagnostic neurosurgical procedure in children with diabetes insipidus (DI) and a thickened pituitary stalk (TPS) on magnetic resonance (MR) imaging. METHODS: Seven children with a TPS who presented with DI eventually underwent surgery for diagnostic purposes. The ages at onset of DI were 6 to 16 years, and the follow-up period until surgery was 26.9 +/- 11.9 months. In four of seven children, the stalk appeared normal on the first MR image, but it was thickened and variably enhancing on later images in all instances. The reason for eventual surgery was endocrinological deterioration in two of seven children, radiological progression in two children, and a combination of the two in three children. Three children experienced visual disturbances and four children had optic nerve, chiasma, or hypothalamus involvement. All children suffered additional endocrinological abnormalities pursuant to the initial DI. A definitive diagnosis was achieved in six of seven children: germinomas in five and Langerhans cell histiocytosis in one. One child had lymphocytic infiltrate. None of the children deteriorated neurologically or endocrinologically after the operation. On follow up, vision deficit was irreversible in all children who demonstrated visual abnormalities before treatment. CONCLUSIONS: Surgery should be performed in children with a TPS and DI for early diagnosis and disease-oriented therapy when there is further endocrinological, radiological, or clinical deterioration. The complication rate is low in open biopsies, and histological diagnosis is achieved in most of the cases. All children who present with central DI must undergo head MR imaging, and even if results are normal, close radiological and clinical follow up is mandatory. PMID- 16370281 TI - Chiari I malformation in patients with FG syndrome. AB - OBJECT: The FG syndrome (FGS) is a common, heterogeneous group of clinically indistinguishable X-linked disorders comprising congenital hypotonia, macrocephaly, psychomotor delay, abnormalities in sensory integration, agenesis of corpus callosum, an unusual personality with behavior abnormalities, and disturbances of gastrointestinal function. On magnetic resonance (MR) imaging, some patients have evidence of tonsillar ectopia. The authors describe the incidence of Chiari I malformation in patients with FGS and attempt to determine the optimal treatment of these patients. METHODS: The authors performed a retrospective chart and radiological review of 144 pediatric patients with FGS for evidence of tonsillar ectopia on brain MR imaging. Eleven (7.6%) of these 144 patients had tonsillar ectopia, and in eight patients (5.6%), the tonsils were located more than 5 mm below the foramen magnum. Four of these patients underwent posterior fossa decompression, and surgery was performed at a mean age of 3 years. Indications for surgery included significant headaches and behavioral problems in two patients and failure to thrive with severe breathing and feeding difficulties in two infants. All four improved after surgery. The other patients remained asymptomatic from their tonsillar ectopia, showed no clinical or radiological signs of progression, and did not require surgery. CONCLUSIONS: Chiari I malformation is more common in individuals with FGS than in the general population. Some of these patients with FGS require decompression surgery, but the decision to operate can be difficult because of their developmental delay, difficulties with language skills, general fatigue, possibility of upper motor neuron dysfunction, behavioral problems, or failure to thrive, which may mask the symptoms of a Chiari I malformation. PMID- 16370282 TI - Pediatric nonvertebral paraspinal arteriovenous fistulas along the segmental nerve: clinical, imaging, and therapeutic considerations. AB - OBJECT: The authors characterize the clinical presentation and imaging features of paraspinal nonvertebral arteriovenous fistulas (AVFs) along the segmental nerve and describe their endovascular treatment. METHODS: The authors undertook a retrospective review of medical records, imaging, and treatment of patients with endovascular problems spanning the period from 1985 to 2003. Five pediatric patients (2-3 years of age) received diagnoses of nonvertebral paraspinal AVFs along the segmental nerve. All patients presented with an incidentally discovered continuous murmur over the paraspinal or parasternal regions. All patients were neurologically intact; two patients had cardiomegaly. The AVF was found in the midthoracic level in four patients and at L-3 in one patient. All AVFs were high flow single-hole fistulas at the neural foramen with venous drainage into paraspinal and epidural veins but without intradural reflux. All fistulas were endovascularly occluded in the same session as diagnostic angiography took place. The fistula was completely occluded, with detachable coils in one case and with N butyl-cyanoacrylate (NBCA) in four cases. Before NBCA injection, the flow through the fistula was decreased either by placing coils distal to the fistula or by inflating a balloon proximally. No signs of recanalization appeared on short-term follow-up magnetic resonance imaging in all patients. All patients remained neurologically intact at the last available follow-up session (mean 6 years). CONCLUSIONS: Nonvertebral paraspinal AVFs along the segmental nerve are specific disease entities seen in children presenting with bruit and cardiomegaly. Endovascular embolization should be the treatment of choice for this rare disease. PMID- 16370283 TI - Effect of 3-tesla magnetic resonance imaging on various pressure programmable shunt valves. AB - OBJECT: This study investigated the changes in the valve pressure setting of several magnetic pressure-programmable valves after exposure to a 3-tesla magnetic field. METHODS: Five each of four types of pressure-programmable shunt valves were tested: Sophy Polaris, Sophy SM8, Codman-Hakim, and Medtronic Strata. First, the valves were advanced toward the 3-tesla static magnetic field. Second, T1-, T2-, and diffusion-weighted magnetic resonance (MR) images were generated with a radiofrequency magnetic field. Any changes in the pressure setting were observed by visual inspection with a compass or radiography. The pressure settings were changed after exposure to the static magnetic field in all programmable valves except for the Sophy Polaris. All pressure settings studied were unchanged after exposure to both static and radiofrequency magnetic fields (T1-, T2-, and diffusion-weighted MR imaging) in the Sophy Polaris. CONCLUSIONS: The Sophy Polaris valve allows shunt-dependent patients who need a programmable valve to undergo 3-tesla MR imaging. PMID- 16370284 TI - Spinal epidural angiolipoma complicated by an intratumoral abscess. Case report. AB - Spinal angiolipomas are rare, benign lesions representing 0.14 to 1.2% of all spinal axis tumors. They most commonly involve the midthoracic spine and are located in the posterior epidural space. Up to now, six pediatric cases have been reported in the literature; two of them involved an acute clinical onset that was related to a venous infarction of a tumor. The authors report the case of a 16 year-old boy with a midthoracic epidural angiolipoma who was admitted with a clinical history of an acute paraparesis. In contrast to previous descriptions, the acute onset in this case was related to a spontaneous intratumoral abscess within the tumor. PMID- 16370285 TI - An unusual presentation of achondroplasia. Case report. AB - The authors present a case of an adolescent with achondroplasia and symptoms of neurogenic claudication who, interestingly, also presented with intermittent priapism. Both the neurogenic claudication and priapism were relieved following a thoracolumbar laminectomy. To our knowledge, this is the first report of a dwarf with thoracolumbar spinal canal stenosis presenting with priapism. PMID- 16370286 TI - Beckwith-Wiedemann syndrome in a child with Chiari I malformation. Case report. AB - The authors present the case of a child diagnosed as having Beckwith-Wiedemann syndrome and Chiari I malformation. Hemihypertrophy is associated with Beckwith Wiedemann syndrome and has been described in conjunction with Chiari I malformation. The authors hypothesize that the hemihypertrophy that may involve the skull base and Chiari I malformation found in their patient are not spurious findings but are pathologically related, perhaps by slight dysmorphologies of the posterior cranial fossa. PMID- 16370287 TI - Acute clival and spinal subdural hematoma with spontaneous resolution: clinical and radiographic correlation in support of a proposed pathophysiological mechanism. Case report. AB - Infratentorial and spinal subdural hematomas (SDHs) from traumatic injury in the pediatric population occur with such rarity that they can present the clinician with a challenge in diagnosis and management. When such hematomas are correctly identified, clinicians must decide whether to evacuate the lesion or manage it expectantly. The authors discuss the case of a 4-year-old child who presented with a clival and spinal SDH after a fall from a fourth-story window. The clinical and radiographic findings support a possible mechanism of evolution of these lesions. There is little evidence to guide management of clival and spinal SDHs. This case supports the evaluation for a spinal SDH when a clival hematoma is diagnosed. In the setting of a good neurological examination, expectant management can be an appropriate method of treatment. Additionally, this case lends insight into the pathophysiology of spinal SDHs. Unlike its intracranial counterpart, the spinal subdural space lacks bridging veins. The mechanism of formation of spinal SDHs after trauma has been heretofore relatively unclear. The images in this case support the hypothesis that redistribution of the clival SDH to dependent areas in the spinal subdural space is a significant mechanism in the evolution of these lesions. PMID- 16370288 TI - Giant intracranial teratoma and lack of cortical development in a fetus. Case report. AB - Antenatal diagnosis of an intracranial neoplasm is extremely rare. The authors describe a case in which a 21-week-old fetus was found, by using fetal ultrasonography, to have a large intracranial mass. Fetal magnetic resonance (MR) images, obtained at 21 and 25 weeks' gestation, supported the diagnosis of a teratoma. As the tumor increased in size, near-complete brain atrophy ensued. Premature labor was induced, and a nonviable fetus died within minutes of delivery. Postmortem analysis confirmed a teratoma occupying a major portion of the intracranial space. In cases in which abnormal brain development is suspected in a fetus, the use of fetal MR imaging can give a clearer picture of the pathological entity, which may allow for a more accurate diagnosis. The usefulness of fetal MR imaging in monitoring brain development and tumor growth during treatment planning is discussed. PMID- 16370289 TI - Neurocytoma-like neoplasm of the thoracic spine in a 15-month-old child presenting with diffuse leptomeningeal dissemination and communicating hydrocephalus. Case report. AB - This unusual mixed glioneuronal neoplasm of the spine resembling central neurocytoma is only the second reported example of a neoplasm of this type involving the spinal cord and is, seemingly, the first to present with diffuse leptomeningeal dissemination and communicating hydrocephalus. This 15-month-old boy presented with somnolence, bilateral sixth nerve palsy, and lower-extremity weakness and was found to harbor a primary neoplasm within the thoracic spine and associated syrinx, widespread leptomeningeal dissemination, and communicating hydrocephalus. The patient underwent cerebrospinal fluid shunt placement, thoracic laminectomy for tumor debulking and biopsy, chemotherapy, and radiation therapy to the neuraxis. Immunohistochemically, the tumor was marked by evidence of mixed glial (glial fibrillary acidic protein-positive) and neuronal (synaptophysin-positive) differentiation. Mitotic activity was inconspicuous. No areas of vascular proliferation, necrosis, or marked nuclear pleomorphism were noted. Mixed glioneuronal neoplasms are a heterogeneous group of tumors whose biological potential remains incompletely defined. The present case illustrates the reality that some of these neoplasms can be clinically aggressive, despite comparatively bland histological features. The authors' goal is to acquaint neurosurgeons with the expanding spectrum of mixed glioneuronal neoplasms and with the potential of some of these lesions to pursue an aggressive clinical course. PMID- 16370290 TI - Transplacental transmission of metastatic melanoma to the posterior fossa. Case report. AB - The authors describe the first documented case of transplacental transmission of metastatic melanoma to the neuraxis. The patient was a 7-month-old boy who presented with signs and symptoms of elevated intracranial pressure. Magnetic resonance imaging revealed an inhomogeneously enhancing posterior fossa mass measuring 5 x 5 x 6 cm and filling the fourth ventricle. A posterior fossa craniotomy was performed. Pathological studies confirmed the presence of a metastatic melanoma that was pathologically identical to that of his mother. The boy received aggressive chemotherapy and underwent an additional resection. He also required a ventriculoperitoneal shunt for treatment of his hydrocephalus. He lived longer than any other patient with transplacental transmission of metastatic melanoma but ultimately died of the disease, 18 months after his initial presentation. PMID- 16370291 TI - Spontaneous regression of syringomyelia in Hajdu-Cheney syndrome with severe platybasia. Case report. AB - Hadju-Cheney syndrome (HCS) is a rare autosomal-dominant disorder with variable expressivity. It is characterized by facial dysmorphism, premature tooth loss, osteolysis of distal phalanges, and skull abnormalities. In some cases, progressive platybasia can occur and can lead to Chiari malformation with an obstruction of cerebrospinal fluid flow. To the best of the authors' knowledge, only five cases of HCS-associated syringomyelia have been reported in the literature. Because of the rarity of this association, little is known about its natural history. The authors present the case of a 16-year-old boy affected by HCS. On initial magnetic resonance (MR) imaging, a severe basilar invagination with Chiari malformation and cervicothoracic syringomyelia was documented. The syringomyelia had no clinical manifestations. A repeated MR image demonstrated a spontaneous resolution of the syrinx with no changes in the tonsil or the platybasia. The regression of the syringomyelia was confirmed by a control MR imaging examination performed after a 2-year period. No changes in the patient's clinical conditions were found during the follow-up period. This is the first case of spontaneous regression of the syringomyelia despite a severe platybasia in HCS. It did not appear correlated to a modification of the tonsil's structure or position. This observation illustrates one possible evolution of syringomyelia in the natural history of HCS and raises the question of the potential mechanisms involved in the spontaneous drainage of the syringomyelic cavity. PMID- 16370292 TI - Progressive syringomyelia controlled by treatment of associated hydrocephalus in an infant with birth injury. Case report. AB - The author's aim in reporting this case is to extend awareness of the importance of management of associated hydrocephalus among patients with syringomyelia to the posttraumatic state. A full-term infant was delivered by cesarean section because of transverse lie. In the 2nd week of life, hypotonia affecting the lower limbs and the left upper limb was recognized. On MR imaging, posterior fossa subdural hemorrhage and spinal cord injury at the T-1 and T-2 levels were evident. Sequential imaging studies demonstrated progressive myelomalacic changes in the cervical spinal cord and eventual development of syrinx cavities as far cephalad as C-3. Hydrocephalus developed simultaneously as well. A ventriculoperitoneal cerebrospinal fluid shunt was inserted at 22 months of age. Six months after shunt insertion, MR imaging demonstrated regression of the posttraumatic syringomyelia. The patient was stable from an imaging standpoint at 16 months after shunt insertion, and she has continued to make developmental progress. The importance of treatment of associated hydrocephalus is widely recognized in the management of syringomyelia associated with the Chiari malformations. It should be considered in the management of posttraumatic syringomyelia as well. PMID- 16370293 TI - Sticky sutures? PMID- 16370294 TI - Occult sacral meningocele. PMID- 16370295 TI - Anterior corpectomy and fusion with fibular strut grafts for multilevel cervical myelopathy. AB - OBJECT: The authors conducted a study to investigate the long-term results and postoperative complications of a new surgical technique, fibular strut graft assisted anterior corpectomy and fusion for multilevel (> four) cervical myelopathy. Multilevel anterior corpectomy and subsequent strut graft placement is considered a challenging procedure because of complications relating to graft dislodgment, pseudarthrosis, greater operative duration, and increased blood loss. METHODS: The study comprised 100 patients with cervical myelopathy who underwent anterior corpectomy and fusion and fibular strut graft placement at more than four disc space levels between 1989 and 1998. Single-screw fixation was used in conjunction with the autologous strut graft. Preoperative and postoperative plain radiographs, computerized tomography myelograms, and magnetic resonance images were obtained for assessment of fusion status. All complications and outcomes were analyzed based on clinical records to evaluate the results of the technique. There were no cases of graft dislodgment. The graft union rate was 85%. Analysis of clinical data showed that pseudarthrosis had no adverse effect on the clinical results. Adjacent-level disc degeneration occurred in 12% of patients, but in all cases the patients were asymptomatic. In 9% of cases C-5 palsy was observed but it recovered spontaneously. There were no infections and no case of neurological deterioration. CONCLUSIONS: With this new graft technique, graft dislodgment, the major complication associated with strut graft surgery, was resolved completely. This simple technique involving single-screw fixation provided good results when used in conjunction with anterior decompression and strut graft fixation with a very low incidence of complications. PMID- 16370296 TI - Anterior spinal decompression and fusion for cervical flexion myelopathy in young patients. AB - OBJECT: The mechanism underlying cervical flexion myelopathy (CFM) is unclear. The authors report the results of anterior decompression and fusion (ADF) in terms of neurological status and radiographically documented status in young patients and discuss the pathophysiological mechanism of the entity. METHODS: Twelve patients underwent ADF in which autogenous iliac bone graft was placed. The fusion area was one segment in four cases, two segments in seven, and three segments in one. Neurological status, as determined by the Japanese Orthopaedic Association (JOA) score, radiographic findings, and intraoperative findings were evaluated. The mean follow-up period was 63.3 months (range 20-180 months). Grip strength was significantly improved and sensory disturbances resolved completely. Intrinsic muscle atrophy, however, persisted in all patients at the final follow up examination. Local kyphosis in the flexed-neck position at the fusion levels was corrected by surgery. Preoperative computerized tomography myelography revealed that the cord compression index, which was calculated by anteroposterior and transverse diameters of the spinal cord, decreased to 33 +/- 6.2% in the flexed-neck position from 39.7 +/- 9.9% in the extended-neck position. The anterior dura mater-spinal cord distance decreased to 1.9 +/- 0.7 mm in the flexed-neck position from 4 +/- 1.2 mm in extended-neck position. The posterior dura mater-spinal cord distance increased to 2.5 +/- 1.1 mm in the flexed-neck position from 1.3 +/- 0.5 mm in the extended-neck position. CONCLUSIONS: Postoperative neurological status was improved in terms of grip strength, sensory disturbance, and JOA score, and local kyphosis in the flexed-neck position at the fusion levels was reduced and stabilized by ADF. In most cases local kyphosis in the flexed-neck position was demonstrated at the corresponding disc level, as were cervical cord compression and decrease of the anterior wall of the dura mater-spinal cord distance in the flexed-neck position. Therefore, the contact pressure between the spinal cord and anterior structures (intact vertebral bodies and intervertebral discs) in the mobile and kyphotic segments was considered to contribute to the onset of CFM. The ADF-related improvement of the clinical symptoms, preventing kyphotic alignment in flexion and decreasing movement of the cervical spine, supports the idea of a contact pressure mechanism. Furthermore, short ADF performed only at the corresponding segments can preserve more mobile segments compared with posterior fusion. Thus, ADF should be the first choice in the treatment of CFM. PMID- 16370297 TI - Palsies of the fifth cervical nerve root after cervical decompression: prevention using continuous intraoperative electromyography monitoring. AB - OBJECT: A desire to prevent complications resulting from spinal surgery led to the development of intraoperative monitoring. Intraoperative electromyography (EMG) provides useful diagnostic information regarding nerve root function during spinal and peripheral nerve surgeries. The C-5 nerve root is considered particularly vulnerable to injury during cervical surgery. Despite advances in techniques, the incidence of postoperative C-5 palsy has not changed. METHODS: The authors reviewed prospectively collected data obtained in 161 patients who underwent 171 cervical procedures. In 116 procedures, operative monitoring was modified to include continuous C-5 EMG from the deltoid muscle. In cases in which spontaneous C-5 activity occurred, an appropriate change in operative manipulation was made. A historical control group consisted of a retrospective review of 55 procedures that were monitored using conventional techniques. In the retrospective cohort, four (7.3%) of 55 patients presented after undergoing surgery for C-5 nerve root palsy. In each patient conventional monitoring revealed unremarkable findings. In the prospective cohort, intraoperative spontaneous EMG activity necessitated a change in either positioning or operative technique in three cases. Only one patient (0.9%) experienced postoperative C-5 palsy. Postoperative C-5 palsy occurred in no patient in whom there was no intra operative evidence of root irritation (p < 0.03, chi-square test). CONCLUSIONS: The incidence of postoperative C-5 palsies was reduced from 7.3% to 0.9% due to intraoperative continuous EMG monitoring. No patient suffered a postoperative C-5 palsy when intraoperative evidence of root irritation was absent. PMID- 16370298 TI - Minimally invasive microendoscopy-assisted transforaminal lumbar interbody fusion with instrumentation. AB - OBJECT: The authors have developed a novel technique for percutaneous fusion in which standard microendoscopic discectomy is modified. Based on data obtained in their cadaveric studies they considered that this minimally invasive interbody fusion could be safely implemented clinically. The authors describe their initial experience with a microendoscopic transforaminal lumbar interbody fusion (METLIF) technique, with regard to safety in the placement of percutaneous instrumentation, perioperative morbidity, and early postoperative results. METHODS: The METLIF procedure was performed unilaterally in 20 patients with single-level lumbar spondylolisthesis or pure mechanical back pain with endoscopic assistance, hemilaminectomy, unilateral facetectomy, and microdiscectomy. Two interbody grafts were placed via the lateral exposure of the disc space. Bilateral percutaneous pedicle screws were then inserted. Compared with patients who had undergone single-level posterior LIF at the same institutions, intraoperative blood loss, hospital length of stay (LOS), and postoperative narcotic agent use were significantly lower in the METLIF group. The mean LOS for the percutaneous fusion group was 3.4 days (5.1 days in those who underwent PLIF; p < 0.02). There have been no procedure-related complications in this series to date. CONCLUSIONS: The METLIF technique provided an option for percutaneous interbody fusion similar to that in open surgery while minimizing destruction to adjacent tissues. This technique was safe and exhibited a trend toward decreased intraoperative blood loss, postoperative pain, total narcotic use, and the risk of transfusion. PMID- 16370299 TI - Bowel and bladder continence, wound healing, and functional outcomes in patients who underwent sacrectomy. AB - OBJECT: Total or partial sacrectomy is a rare procedure in patients with locally invasive tumors involving the sacrum; it may be associated with functional loss, such as bowel and bladder dysfunction and gait abnormality. In this study the authors examined functional outcome following sacrectomy. METHODS: The authors reviewed the charts of 50 consecutive patients who had undergone sacrectomy between July 1993 and August 2002. There were 23 male and 27 female patients whose mean age was 46 years (range 13-86 years). Twelve patients with rectal cancer underwent a separate analysis. The patients without rectal cancer were divided into two groups: those who had undergone colostomy for bowel diversion (Group 1, six cases), and those who had not (Group 2, 32 cases). In Group 1 patients the median hospital length of stay (LOS) was 48.5 days (the 25th% and 75th percentiles are 26 and 58, respectively), and in Group 2 patients the median LOS was 18.5 days (the 25th and 75th percentiles are 8 and 41, respectively; p = 0.14). In Group 2 (non-rectal cancer without colostomy), LOS was greater in patients in whom a myocutaneous flap was used compared with those in whom no flap was used (36 days compared with 8.5 days, respectively; p = 0.0012); in patients with bowel incontinence the median LOS was significantly longer than that in patients with bowel continence (39 days compared with 8 days, respectively; p = 0.0026). The incidence of bowel incontinence in Group 2 was closely related to the integrity of the S-3 nerve root (p = 0.05). CONCLUSIONS: Awareness of the association between S-3 nerve root resection and bowel and bladder incontinence may help surgeons' decision-making process. PMID- 16370300 TI - En bloc resection of primary sacral tumors: classification of surgical approaches and outcome. AB - OBJECT: En bloc resection with adequate margins is associated with the highest probability of long-term tumor control or cure in most cases of primary sacral malignancies. The authors present their experience with a systematic approach to these lesions. They provide a novel classification of surgical techniques based on the level of nerve root sacrifice and evaluate the functional and oncological outcomes. METHODS: Seventy-eight consecutive patients underwent 94 resections of sacral neoplasms at The University of Texas M. D. Anderson Cancer Center in Houston between August 1993 and June 2002. The records of 29 consecutive patients who underwent en bloc resection of primary sacral tumors were retrospectively reviewed. The median follow-up period was 55 months (range 1-103 months). Chordoma was the most frequent tumor type (16 cases). Midline sacral amputation was performed in 25 patients (eight low, four middle, seven high, and five total sacrectomies; one hemicorporectomy). Lateral sacrectomy was undertaken in four patients (two unilateral excisions of the sacroiliac joint and two hemisacrectomies). The surgical margins were wide in 19 cases, marginal in nine, and contaminated in one. The type of sacrectomy correlated with characteristic outcomes with respect to bladder, bowel, and ambulatory functions. Duration of hospital stay was related to the extent of sacrectomy (p = 0.003, Wilcoxon signed rank test). The median Kaplan-Meier disease-free survival for patients with chordoma was 68 months (95% confidence interval 46-90 months). CONCLUSIONS: Classification of en bloc sacral resection techniques by the level of nerve root transection is useful in predicting postoperative function and the potential for morbidity. Adequate surgical margins should not be compromised to preserve function when they are necessary to affect tumor control. PMID- 16370301 TI - Anterolateral approach to the craniocervical junction. AB - OBJECT: The authors present the surgical results obtained using the anterolateral approach to the craniocervical junction (CCJ) to resect a lesion with an extradural component located anterolateral to the foramen magnum and upper cervical spine. METHODS: The anterolateral approach, which is a presternomastoid retrojugular route to the CCJ, was performed in 14 patients. The skin incision follows the anterior edge of the sternomastoid muscle. The vertebral artery (VA) was exposed at C-1. This approach was extended either down to the cervical spine or anteriorly to the jugular foramen, according to specific requirements. Two patients had previously undergone other surgical procedures. The follow-up period ranged from 4 months to 6.2 years. The tumor resection was complete in 11 cases and subtotal in two. In a case of vertebral coiling, a vein graft was interposed between the V1 and the V3 segments of the VA, and the bypass was still patent at the 2-year follow-up examination. In two cases involving a glomus tumor, there was a transitory postoperative seventh cranial nerve deficit. CONCLUSIONS: The aforementioned technique allows for sufficient access to lesions located anterolateral to the CCJ. It is indicated in cases in which lesions exhibit a significant extradural component, and it provides good control of the VA, the cervical portion of the internal carotid artery, sigmoid-jugular complex, and lower cranial nerves. This approach can easily be combined with a posterolateral approach and can be extended anteriorly toward the jugular foramen and inferiorly toward the lower cervical spine. PMID- 16370302 TI - Outcome after less-invasive decompression of lumbar spinal stenosis: a randomized comparison of unilateral laminotomy, bilateral laminotomy, and laminectomy. AB - OBJECT: Recently, limited decompression procedures have been proposed in the treatment of lumbar stenosis. The authors undertook a prospective study to compare the safety and outcome of unilateral and bilateral laminotomy with laminectomy. METHODS: One hundred twenty consecutive patients with 207 levels of lumbar stenosis without herniated discs or instability were randomized to three treatment groups (bilateral laminotomy [Group 1], unilateral laminotomy [Group 2], and laminectomy [Group 3]). Perioperative parameters and complications were documented. Symptoms and scores, such as visual analog scale (VAS), Roland-Morris Scale, Short Form-36 (SF-36), and patient satisfaction were assessed preoperatively and at 3, 6, and 12 months after surgery. Adequate decompression was achieved in all patients. The overall complication rate was lowest in patients who had undergone bilateral laminotomy (Group 1). The minimum follow up of 12 months was obtained in 94% of patients. Residual pain was lowest in Group 1 (VAS score 2.3 +/- 2.4 and 4 +/- 1 in Group 3; p < 0.05 and 3.6 +/- 2.7 in Group 2; p < 0.05). The Roland-Morris Scale score improved from 17 +/- 4.3 before surgery to 8.1 +/- 7, 8.5 +/- 7.3, and 10.9 +/- 7.5 (Groups 1-3, respectively; p < 0.001 compared with preoperative) corresponding to a dramatic increase in walking distance. Examination of SF-36 scores demonstrated marked improvement, most pronounced in Group 1. The number of repeated operations did not differ among groups. Patient satisfaction was significantly superior in Group 1, with 3, 27, and 26% of patients unsatisfied (in Groups 1, 2, and 3, respectively; p < 0.01). CONCLUSIONS: Bilateral and unilateral laminotomy allowed adequate and safe decompression of lumbar stenosis, resulted in a highly significant reduction of symptoms and disability, and improved health-related quality of life. Outcome after unilateral laminotomy was comparable with that after laminectomy. In most outcome parameters, bilateral laminotomy was associated with a significant benefit and thus constitutes a promising treatment alternative. PMID- 16370303 TI - A novel intramedullary spinal cord tumor model: functional, radiological, and histopathological characterization. AB - OBJECT: Survival rates for high-grade intramedullary spinal cord tumors (IMSCTs) are approximately 30%, and optimal therapy has yet to be determined. Development of a satisfactory intramedullary tumor model is necessary for testing new therapeutic paradigms that may prolong survival. The authors report the technique, functional progression, radiological appearance, and histopathological features of a novel intramedullary model in rabbits. METHODS: Ten New Zealand white rabbits were randomized to receive an intramedullary injection of either 25 microl of VX2 carcinoma cells (500,000 cells; six rabbits) or 25 microl of medium (Dulbecco modified Eagle medium; four rabbits) into the midthoracic spinal cord. Postoperatively the rabbits were evaluated twice daily for neurological deficits. High-resolution magnetic resonance (MR) images were acquired preoperatively and weekly postoperatively until onset of paraparesis, at which point the animals were killed, and the midthoracic spines were processed for histopathological examination. The VX2-carcinoma cells grew in 100% of animals injected and resulted in a statistically significant mean onset of paraparesis of 16.8 +/- 1.7 days (p = 0.0035, log-rank test), compared with animals in the control group in which neurological deficits were absent by Day 45. Contrast-enhanced T1-weighted MR imaging best demonstrated space-occupying intramedullary lesions and histopathological findings confirmed the intramedullary location of the tumor. Animals in the control group exhibited no functional, radiographic, or pathological signs of tumor. CONCLUSIONS: Progression to paraparesis was consistent in all the VX2-injected animals, with predictable onset of paraparesis occurring approximately 17 days postinjection. Histopathological and radiological characteristics of the VX2 intramedullary tumor are comparable with those of aggressive primary human IMSCTs. Establishment of this novel animal tumor model will facilitate the testing of new therapeutic paradigms for the treatment of IMSCTs. PMID- 16370304 TI - The filum terminale externum. AB - OBJECT: The filum terminale externum (FTE) is the extradural component of the filum terminale internum and little attention has been dedicated to this structure in the literature. The authors theorized that the rare intrasacral ependymomas may originate from ependymal cell collections within the FTE. METHODS: To address this hypothesis, the FTE was dissected and analyzed histologically in 15 adult cadavers. None of the specimens was found to harbor ependymal or other glial cell collections. CONCLUSIONS: The authors found previously undescribed smooth-muscle cells within the FTE. Furthermore, histological analysis identified adipose, nerve, bone, and cartilage cells. PMID- 16370305 TI - An atypical spinal meningioma with CSF metastasis: fatal progression despite aggressive treatment. Case report. AB - The authors report the case of a 23-year-old man who presented with a C1-3 spinal mass. Following intraspinal decompression the tumor was histologically classified as an atypical meningioma (World Health Organization grade II). Two further surgical interventions resulted in almost total removal of the meningioma. In addition, radiotherapy was performed. During the 1.5-year follow-up period the diagnostic examinations identified a local tumor recurrence, an intraspinal C-6 metastasis, and a segmental instability with anterior C2-3 slippage and C3-4 kyphosis. The tumor was resected and occipitocervical stabilization was performed. Histological examination showed no change in malignancy. Despite additional hydroxyurea-based chemotherapy, the patient presented 4 months later with a hemiparesis and a massive recurrence of the tumor mass involving the posterior fossa and the upper thoracic spine. Because there were no further therapeutical options, the patient died. The authors discuss more aggressive therapeutic options in addition to surgery in patients with metastatic atypical meningiomas. The results in the reported case indicate that meningiomas associated with cerebrospinal fluid metastasis may represent a higher grade of malignancy. PMID- 16370306 TI - Osteoradionecrosis of the cervical spine resulting from radiotherapy for primary head and neck malignancies: operative and nonoperative management. Case report. AB - Osteoradionecrosis is a process of dysvascular bone necrosis and fibrous replacement following exposure to high doses of radiation. The poorly vascularized necrotic tissue may cause pain and/or instability, and it cannot resist infection well, which may result in secondary osteomyelitis. When these processes affect the cervical spine, the resulting instability and neurological deficits can be devastating, and immediate reestablishment of spinal stability is paramount. Reconstruction of the cervical spine can be particularly challenging in this subgroup of patients in whom the spine is poorly vascularized after radical surgery, high-dose irradiation, and infection. The authors report three cases of cervical spine osteoradionecrosis following radiotherapy for primary head and neck malignancies. Two patients suffered secondary osteomyelitis, severe spinal deformity, and spinal cord compression. These patients underwent surgery in which a vascularized fibular graft and instrumentation were used to reconstruct the cervical spine; subsequently hyperbaric oxygen (HBO) therapy was instituted. Fusion occurred, spinal stability was restored, and neurological dysfunction resolved at the 2- and 4-year follow-up examinations, respectively. The third patient experienced pain and dysphagia but did not have osteomyelitis, spinal instability, or neurological deficits. He underwent HBO therapy alone, with improved symptoms and imaging findings. Hyperbaric oxygen is an essential part of treatment for osteoradionecrosis and may be sufficient by itself for uncomplicated cases, but surgery is required for patients with spinal instability, spinal cord compression, and/or infection. A vascularized fibular bone graft is a very helpful adjunct in these patients because it adds little morbidity and may increase the rate of spinal fusion. PMID- 16370307 TI - Image-guided surgery for thoracic ossification of the posterior longitudinal ligament. Technical note. AB - The authors describe an anterior decompression procedure for thoracic ossification of the posterior longitudinal ligament (PLL) in which they used an image guidance system in three cases. To make registration possible in anterior thoracic surgery, they devised a surgical reference frame that could be connected to a rod and attached to an external fixation device, which was then attached to the thoracic VB. The mean fiducial error at the registration was acceptable (range 0.5-0.8 mm). They were able to confirm the success of decompression on postoperative computerized tomography scans. In the removal of an ossified thoracic PLL, an image guidance system has been shown to be a useful tool. PMID- 16370308 TI - Cervical instability. PMID- 16370309 TI - Correlation study of toxicity of substituted phenols to river bacteria and their biodegradability in river water. AB - OBJECTIVE: To study the correlation of toxicity with biodegradability (BODT) in order to promote QSBR development and understand the degradation mechanism. METHODS: Toxicity of substituted phenols to river bacteria was determined by the turbidities that were measured using a spectrophotometer (UV-190) at 530 nm against a blank control. The biodegradability of substituted phenols was expressed as BODT and the DO concentrations were determined by the iodometric titration method. RESULTS: The BODT and toxicity(log 1/IC50) of 12 substituted phenols to bacteria from the Songhua River were determined respectively. The correlation of biodegradability with toxicity was developed: BODT=8.21 (+/-2.22) pKa -32.44 (+/-8.28) log 1/IC50 +89.04 (+/-38.20), n=12, R2=0.791, R2(adj)=0.745, SE=9.134, F=17.066, P=0.001. CONCLUSION: The BODT of substituted phenols was influenced by their toxicity and the ionization constant pKa. The stronger the toxicity, the less readily the compound was degraded by river bacteria. PMID- 16370310 TI - Role of cyclinD1 and CDK4 in the carcinogenesis induced by silica. AB - OBJECTIVE: To study the role of cyclinD1 and CDK4 in malignant transformation of human fetal lung diploid fibroblast cell line (2BS) induced by silica. METHODS: Recombination vectors with sense and antisense pXJ41-cyclinD1 and pXJ41-CDK4 were constructed, and then transfected into the malignant transformed cells induced by silica, respectively. At the same time, pXJ41-neo was used as the control. RESULTS: During the progress of the malignant transformation of 2BS cells induced by silica, cyclinD1 and CDK4 were overexpressed. Antisense RNA suppressed cyclinD1 and CDK4 gene expression in the antisense pXJ41-cyclinD1 and pXJ41-CDK4 transfected cells. Antisense RNA led to cell cycle arrest, resulting in lengthened G1 phase (the percentages of cells in the G1 phase changed from 45.1% to 52.7% and 58.0% for cyclinD1 and CDK4 transfected cells, respectively), and eventually attenuated the increase of the proliferation of malignant transformed cells induced by silica. Compared with malignant transformed cells induced by silica, cells transfected with antisense pXJ41-cyclinD1 and pXJ41-CDK4 showed obviously reduced growth rates. On the 8th day, the suppression rates were 58.69 and 77.43% (the growth rate of malignant transformed cells induced by silica was 100%), doubling time changed from 21.0 h to 31.4 h and 21.0 h to 42.7 h, respectively, the growth capacities on soft agar of cells transfected by antisense pXJ41-cyclinD1 and pXJ41-CDK4 decreased obviously. CONCLUSION: CyclinD1 and CDK4 play an important role in maintaining transformed phenotype of the cancer cells. PMID- 16370311 TI - Studies of chemical constituents and their antioxidant activities from Astragalus mongholicus Bunge. AB - OBJECTIVE: To evaluate the antioxidant activities of different chemical constituents from Astragalus mongholicus Bunge and their protection against xanthine (XA)/xanthine oxidase (XO)-induced toxicity in PC12 cells. METHODS: The compounds of Astragalus mongholicus Bunge were isolated by chromatography and the structures were elucidated on the basis of spectral data interpretation. Their antioxidant activities were detected by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities in a cell-free system. Meanwhile, the effects against XA/XO-induced toxicity were assessed using MTT assay in PC12 cells. RESULTS: Ten principal constituents were isolated and identified as formononetin (I), ononin (II), calycosin (III), calycosin-7-O-beta-D-glucoside (IV), 9,10 dimethoxypterocarpan-3-O-beta-D-glucoside (V), adenosine (VI), pinitol (VII), daucosterol (VIII), beta-sitoster (IX) and saccharose (X) from Astragalus mongholicus Bunge. The compounds I, III, and IV scavenged DPPH free radicals in vitro. Formononetin and calycosin were found to inhibit XA/XO-induced cell injury significantly, with an estimated EC50 of 50 ng/mL. CONCLUSION: Compound II, VI, and VII are first reported in this plant. Calycosin exhibits the most potent antioxidant activity both in the cell-free system and in the cell system. PMID- 16370312 TI - Over-expressed genes detected by suppression subtractive hybridization in carcinoma derived from transformed 16HBE cells induced by BPDE. AB - OBJECTIVE: To screen the over differentially expressed genes in carcinoma induced by BPDE-transformed 16HBE cells (16HBE-C cells). METHODS: The suppression subtractive hybridization (SSH) method was performed to profile differentially expressed genes between 16HBE-C cells and 16HBE cells. The cDNA fragments of differentially expressed genes were inserted into TA cloning vector and transformed competent E. coli strain. Positive clones were randomly picked up and identified by the colony PCR method. Dot blot was used to test the same source with the tester. The differentially expressed cDNA fragments were sequenced and compared with known genes and EST database in Genbank. RESULTS: Eight known genes were over-expressed in 16HBE-C cells including eukaryotic translation elongation factor 1 alpha 1, HIF-1 responsive RTP801, ribosomal protein L10 (RPL10), ribosomal protein S29 (RPS29), mitochondrion related genes, and laminin receptor 1. Three differentially expressed cDNA fragments could not be matched to the known genes but to the EST database. CONCLUSION: The SSH method can detect differentially expressed genes between 16HBE-C and 16HBE cells. BPDE-induced carcinogenesis may be related to alteration of at least eight known genes and three unknown genes. These expression data provide a clue to further cloning novel genes and studying functions in BPDE-induced carcinoma. PMID- 16370313 TI - Testing potential effect of environmental endocrine disruptors in cow milk on reproductive index in female rats. AB - OBJECTIVE: To study the effect of endocrine disruptor chemicals in cow milk on female reproductive system. METHODS: A two-generation reproduction was conducted according to U. S. FDA standard. Milk was fed in special bottle to Wistar rats of both sexes through two successive generations (F0 and F1) in the milk group while artificial milk was fed to rats in the control group. Twenty-four rats of each sex were mated in each group. Measurements were made according to this guideline. RESULTS: Reproductive parameters in the milk group such as fertility index, gestation index, weights of uterus and ovary, days of vaginal opening, estrous cycles, histological morphological changes were comparable to those in the control group. However, the means of body weight had some differences. The body weight gains increased significantly in the milk-treated group in F1 and F2 generation compared with those in the control group. The concentration of insulin like growth factor-1 (IGF-1) in blood in the milk group was comparable to that in the control group, but the standard deviation changed greatly in the milk-treated rats. CONCLUSION: Endocrine disruptor chemicals in milk have no severe effects on the female reproductive system. PMID- 16370314 TI - Inhibitory effects of alpha-zearalenol on angiotensin II-induced integrin beta3 mRNA via suppression of nuclear factor-kappaB. AB - OBJECTIVE: To investigate the effect of alpha-zearalenol on angiotensin II induced beta3 integrin mRNA expression in human umbilical vein endothelial cells (HUVECs). METHODS: The mRNA level in integrin beta3 was determined by reverse transcription-polymerase chain reaction. Endothelial NF-kappaB activity was determined by the luciferase activity assay of plasmid NF-kappaB-LUC. RESULTS: The angiotensin II-induced beta3 integrin mRNA expression was inhibited by alpha zearalenol and 17beta-estradiol (10 nmol/L -1 micromol/L), but not influenced by ICI 182, 780, a pure competitive antagonist for estrogen receptor or a nitric oxide inhibitor Nomega-Nitro-L-arginine methyl ester hydrochloride. Alpha zearalenol and 17beta-estradiol suppressed the angiotensin II-induced activation of NF-kappaB in endothelial cells. CONCLUSION: Alpha-zearalenol inhibits angiotensin II-induced integrin beta3 mRNA expression by suppressing NF-kappaB activation in endothelial cells. PMID- 16370315 TI - Preparation of monoclonal antibody against HPT and its application to detecting marker protein in genetically modified rice. AB - OBJECTIVE: To produce the monoclonal antibodies (mAbs) against hygromycin B phosphotransferase (HPT) and to develop immunoassay based on mAbs for biosafety assessment of HPT in genetically modified rice (GM rice). METHODS: BALB/c mice were immunized with purified recombinant 6His. HPT protein, and the conventional hybridoma technology was used to generate the monoclonal hybridoma cells. ELISA and Western blot were used to analyze the specificity of mAbs recognizing HPT and the cross reaction with other proteins. A double-Ab sandwich ELISA method was established to detect HPT expression level in the sck gene-modified rice plants. RESULTS: Four hybridomas, named F1, D4-2, D4-4, and D4-5, producing the mAbs against HPT were successfully obtained with the titer of ascetic mAbs ranging from 1x10(-4) to 1x10(-5). Identification of subclass showed that all the produced mAbs belonged to IgG1. Western blot showed specific binding reaction between the mAbs to the HPT proteins expressed in the GM rice. A double sandwich ELISA coated with anti-HPT polyclonal antibody was established with mAbs as sandwich antibody, which showed a sensitivity of 30ng/mL and did not crossreact with other proteins. The expression level of HPT in the leaves of sck-transformed lines was detected (80-150ng/mL). But HPT protein in the grain and seed of GM rice could not be detected using this ELISA assay. CONCLUSION: Anti-HPT mAbs prepared herein have a high specificity and can be used for rapid assay of HPT antigen. The expression level of HPT in the GM rice grain and seed is lower than our ELISA detection limit. PMID- 16370317 TI - Autism and mental retardation of young children in China. AB - OBJECTIVE: To understand the prevalence and rehabilitation status of autism and mental retardation in China. METHODS: Screening test and clinical assessment were conducted for the diagnosis of autism and mental retardation. The assessment included investigation of the histories of medical conditions and development of these two disorders, utilization and needs for the rehabilitation service, and related intellectual and behavioral appraisal. RESULTS: Among the 7345 children investigated, the prevalence of autism disorder was 1.10 cases per 1000 children aged 2-6 years (95% CI=0.34 to 2.54), and the prevalence of mental retardation was 10.76 cases per 1000 children (95% CI=8.40 to 13.12). All the children suffering from autistic disorder were intellectually disabled, whereas 31.0% of the non-autism mental retardates had other disabilities. The medical conditions prior to birth and perinatal period were important potential factors for autism. Half of the autistic children and 84% of the children with non-autism mental retardation had never received any rehabilitative service. CONCLUSIONS: The prevalence of autistic disorder in children aged 2-6 years in Tianjin is rather high. It is urgent to improve the status of the autistic and intelligently disabled young children in China. In order to upgrade the level of early diagnostic and improve the intervention to autism and mental retardation, public awareness and training courses should be heightened. PMID- 16370316 TI - Study on the adoption of Schwann cell phenotype by bone marrow stromal cells in vitro and in vivo. AB - OBJECTIVE: To explore the possibilities of bone marrow stromal cells (MSCs) to adopt Schwann cell phenotype in vitro and in vivo in SD rats. METHODS: MSCs were obtained from tibia and femur bone marrow and cultured in culture flasks. Beta mercaptoethanol followed by retinoic acid, forskolin, basic-FGF, PDGF and heregulin were added to induce differentiation of MSCs'. Schwann cell markers, p75, S-100 and GFAP were used to discriminate induced properties of MSCs' by immunofluorescent staining. PKH-67-labelled MSCs were transplanted into the mechanically injured rat sciatic nerve, and laser confocal microscopy was performed to localize the PKH67 labelled MSCs in the injured sciatic nerve two weeks after the operation. Fluorescence PKH67 attenuation rule was evaluated by flow cytometry in vitro. RESULTS: MSCs changed morphologically into cells resembling primary cultured Schwann cells after their induction in vitro. In vivo, a large number of MSCs were cumulated within the layer of epineurium around the injured nerve and expressed Schwann cell markers, p75, S-100, and GFAP. CONCLUSION: MSCs are able to support nerve fiber regeneration and re-myelination by taking on Schwann cell function, and can be potentially used as possible substitutable cells for artificial nerve conduits to promote nerve regeneration. PMID- 16370318 TI - Comparative proteomic analysis of B. henselae Houston and B. henselae Marseille by two-dimensional gel electrophoresis. AB - OBJECTIVE: To compare the protein difference between B. henselae Houston and B. henselae Marseille by two-dimensional gel electrophoresis. METHOD: Protein samples were prepared by vorterx, ultrasonic treatment, and centrifugation. Protein concentrations were determined by Bradford method. Protein difference was compared by the first IEF and the second SDS-polyacrylamide gel electrophoresis. RESULTS: Protein concentrations in samples of Bartonella henselae Houston and Bartonella henselae Marseille were 2.117 microg/microL and 2.200 microg/microL respectively. Sample protein of 40 microg for IPG strips loading was perfect. The results of 2-DE in pH 4 to 7 IPG strips showed that the total protein spots of Bartonella henselae Houston and Bartonella henselae Marseille were 375 and 379 respectively, 95% of the spots were the same between the two strains of Bartonella henselae. CONCLUSION: The procedure of 2-DE may prove successful for the proteomic analysis of Bartonella henselae. Bartonella henselae Houston and Bartonella henselae Marseille are different genotypes. PMID- 16370319 TI - Study on model of indoor air pollution forecast for decoration under natural ventilation condition. AB - OBJECTIVE: To establish the model of indoor air pollution forecast for decoration. METHODS: The model was based on the balance model for diffusing mass. RESULTS: The data between testing concentration and estimating concentration were compared. The maximal error was less than 30% and average error was 14.6%. CONCLUSION: The model can easily predict whether the pollution for decoration exceeds the standard and how long the room is decorated. PMID- 16370320 TI - Health effects of airborne particulate matter trace elements. AB - The effects of airborne particulate matter (PM) trace elements on health are widely concerned nowadays. Many achievements have been made while many unknowns exist. This article reports the recent research progresses, describes the effects of exposure to PM trace elements on health epidemiological evidence, toxicology findings, and raises some questions for future studies. PMID- 16370321 TI - Considerations in applying the general equilibrium approach to environmental health assessment. AB - There are currently two commonly used approaches to assessing economic impacts of health damage resulting from environmental pollution: human capital approach (HCA) and willingness-to-pay (WTP). WTP can be further divided into averted expenditure approach (AEA), hedonic wage approach (HWA), contingent valuation approach (CVA) and hedonic price approach (HPA). A general review of the principles behind these approaches by the authors indicates that these methods are incapable of unveiling the mechanism of health impact from the point of view of national economy. On a basis of economic system, the shocks brought about by health effects of environmental pollution change the labor supply and medical expenditure, which in turn affects the level of production activity in each sector and the total final consumption pattern of the society. The general equilibrium approach within the framework of macroeconomic theory is able to estimate the health impact on national economy comprehensively and objectively. Its mechanism and applicability are discussed in detail by the authors. PMID- 16370322 TI - Factors involved in self-renewal and pluripotency of embryonic stem cells. AB - Embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of blastocyst stage embryos. These cells possess two unique characteristics: an indefinite self-renewal capacity and pluripotency, the ability to differentiate to cells from the three germ layers. Both human and mouse ES cells are currently at the center of intensive research. One of the burning issues in this research is the way in which these cells remain undifferentiated and maintain their pluripotency. In the past years, data has accumulated concerning the pathways responsible for the unique phenotype of these cells, in both human and mouse. This paper will review the main extrinsic factors and intrinsic transcriptional pathways currently implicated in the self-renewal and pluripotency of ES cells. PMID- 16370323 TI - Embryonic stem cell-derived embryoid bodies: an in vitro model of eutherian pregastrulation development and early gastrulation. AB - In this review, I describe the dawn of embryoid body research and the influence of stem cell properties on embryoid body development. I will focus on the in vitro differentiation of embryonic stem cells in embryoid bodies. I summarize and combine published data for embryo-like development of embryoid bodies, and based on these findings, I will discuss open questions, concerns, and possible future directions of this still emerging field of research. I hope to provide new perspectives and experimental approaches that go beyond the current state of the art to foster an understanding of eutherian embryogenesis and provide clues for the efficient production of somatic cells for cell therapy. PMID- 16370324 TI - Embryonic stem cells: a novel tool for the study of antiangiogenesis and tumor induced angiogenesis. AB - Major research initiatives in antiangiogenesis research have been undertaken to control angiogenic diseases such as polyarthritis, psoriasis, endometriosis, and diabetic retinopathy, and inhibition of tumor-induced angiogenesis has emerged as one of the most promising anti-cancer therapies currently available. Although several quantitative in vivo (i.e., animal models) as well as in vitro (i.e., pure endothelial cell cultures) angiogenesis assays have been described, the development of novel angiogenesis assays with organotypic culture systems that take into account oxygen and nutrient gradients, depth-dependent changes in intracellular pH and a redox state similar to that found in a natural tissue microenvironment are necessary to investigate blood vessel growth. Embryonic stem cells of mouse and human origin have the capacity to develop into three dimensional tissues with functional capillaries, and this model system represents an excellent in vitro model for antiangiogenesis research. Upon confrontation of stem cells by co-culture with multicellular tumor spheroids, tumor-induced angiogenesis, i.e., the invasion of endothelial host-derived cells into a tumor tissue, can also be monitored. The current review provides an overview of embryonic stem cells as novel tools for antiangiogenesis research and outlines the use of confrontation cultures for the study of tumor-induced angiogenesis. PMID- 16370326 TI - Cardiomyocytes from human embryonic stem cells. AB - Terminal heart failure is characterized by a significant loss of cardiac myocytes. Stem cells represent a possibility for replacing these lost myocytes but the question of which stem cells are most ideally suited for cell transplantation therapies is still being addressed. Here, we consider human embryonic stem cells (HESC), derived from human embryos in this context. We review the methods used to induce their differentiation to cardiomyocytes in culture, their properties in relation to primary human cardiomyocytes and their ability to integrate into host myocardium. In addition, issues regarding their safety that need addressing before use in cell transplantation therapies, both generally and specifically in relation to the heart, are considered. PMID- 16370325 TI - Cardiac repair by embryonic stem-derived cells. AB - Cell transplantation approaches offer the potential to promote regenerative growth of diseased hearts. It is well established that donor cardiomyocytes stably engraft into recipient hearts when injected directly into the myocardial wall. Moreover, the transplanted donor cardiomyocytes participate in a functional syncytium with the host myocardium. Thus, transplantation of donor cardiomyocytes resulted in at least partial restoration of lost muscle mass. It is also well established that embryonic stem (ES) cells differentiate into cells of ecto-, endo-, and mesodermal lineages when cultured under appropriate conditions in vitro. Robust cardiomyogenic differentiation was frequently observed in spontaneously differentiating ES cultures. Cellular, molecular and physiologic analyses indicated that ES-derived cells were bona fide cardiomyocytes, with in vitro characteristics typical for cells obtained from early stages of cardiac development. Thus, ES-derived cardiomyocytes constitute a viable source of donor cells for cell transplantation therapies. PMID- 16370327 TI - Autocrine and intracrine signaling for cardiogenesis in embryonic stem cells: a clue for the development of novel differentiating agents. AB - Cardiogenesis, one of the earliest and most complex morphogenetic events in the embryo, is not fully understood at the molecular level and is typically a low yield process. Affording a high throughput of cardiogenesis from a suitable population of pluripotent cells is therefore a major assignment in the perspective of a stem cell therapy for heart failure. Analysis of cardiac lineage commitment in mouse embryonic stem cells and in vivo models of cardiac differentiation revealed that a number of crucial growth factors are released from precursor cells, acting in an autocrine fashion on specific plasma membrane receptors to prime a cardiogenic decision. Nevertheless, it is increasingly becoming evident that cell nuclei harbor the potential for intrinsic signal transduction pathways. The term "intracrine" has been proposed for growth regulatory peptides that have been shown to act within their cell of synthesis at the level of the nuclear envelope, chromatin, or other subnuclear components. Considerable evidence links known intracrines with transcriptional responses and self-sustaining loops that behave as long-lived signals and impart features characteristic of differentiation, growth regulation and cell memory. This review focuses on a number of autocrine and intracrine systems within the context of cardiac differentiation and emphasizes the identification of cardiogenic mechanisms as a clue for the development of unprecedented differentiating strategies. In this regard, recently synthesized mixed esters of hyaluronan with butyric and retinoic acid primed the expression of cardiogenic genes and elicited a remarkable increase in cardiomyocyte yield in mouse embryonic stem cells. This demonstrates the potential for chemically modifying the gene program of cardiac differentiation without the aid of gene transfer technologies and sets the basis for the design of a novel generation of chemicals suited for the organization of targeted lineage patterning in stem cells. PMID- 16370328 TI - Therapeutic potential of stem cells in diabetes. AB - Stem cells possess the ability to self-renew by symmetric divisions and, under certain circumstances, differentiate to a committed lineage by asymmetric cell divisions. Depending on the origin, stem cells are classified as either embryonic or adult. Embryonic stem cells are obtained from the inner cell mass of the blastocyst, a structure that appears during embryonic development at day 6 in humans and day 3.5 in mice. Adult stem cells are present within tissues of adult organisms and are responsible for cell turnover or repopulation of tissues under normal or exceptional circumstances. Taken together, stem cells might represent a potential source of tissues for cell therapy protocols, and diabetes is a candidate disease that may benefit from cell replacement protocols. The pathology of type 1 diabetes is caused by the autoimmune destruction or malfunction of pancreatic beta cells, and consequently, a lack of insulin. The absence of insulin is life-threatening, thus requiring diabetic patients to take daily hormone injections from exogenous sources; however, insulin injections do not adequately mimic beta cell function. This results in the development of diabetic complications such as neuropathy, nephropathy, retinopathy and diverse cardiovascular disorders. This chapter intends to summarize the possibilities opened by embryonic and adult stem cells in regenerative medicine for the cure of diabetes. PMID- 16370329 TI - Markers of adult tissue-based stem cells. AB - The expectation generated by the pluripotentiality of embryonic stem (ES) cells has initiated a renaissance in stem cell biology. While ES cells can be harvested in abundance and appear to be the most versatile of cells for regenerative medicine, adult stem cells also hold promise, but the identity and subsequent isolation of these comparatively rare cells remains problematic in most tissues, perhaps with the notable exception of the bone marrow. Identifying surface molecules (markers) that would aid in stem cell isolation is thus a major goal for stem cell biologists. Moreover, the characterization of normal stem cells in specific tissues may provide a dividend for the treatment of cancer. There is a growing belief that the successful treatment of neoplastic disease will require specific targeting of the cancer stem cells, cells that may well have many of the characteristics of their normal counterparts. PMID- 16370330 TI - Designer cytokines for human haematopoietic progenitor cell expansion: impact for tissue regeneration. AB - Haematopoietic stem cells after bone marrow transplantation can expand 100-fold and thereby completely reconstitute the haematopoietic system. Such an expansion has so far not been achieved in vitro using cytokines and growth factors. An interesting parallel exists between haematopoietic stem cells and mouse embryonic stem cells. In both cell types, massive gp130 stimulation results in an inhibition of cellular differentiation. For both cell types, several intrinsic transcription factors are known which, when overexpressed, lead to massive self renewal of the cells. It is currently unknown how in haematopoietic stem cells these intrinsic transcription factors, which include HOXB4 and Bmi-1, can be stimulated by extrinsic signals. Once such signals have been identified, they will be combined with the hyperstimulation of gp130 using designer cytokines. Additional strategies include a cell-permeable version of the HOXB4 protein and members of the WNT family ofligands. These strategies might eventually lead to an expansion of haematopoietic stem cells to a similar extent as that observed after bone marrow transplantation. Should haematopoietic stem cell expansion prove feasible and safe, there is great potential for the treatment of leukaemic disorders but also for the treatment of less severe diseases and non-malignant genetic disorders. PMID- 16370331 TI - Mesenchymal stem cells: isolation, in vitro expansion and characterization. AB - Mesenchymal stem cells (MSC), one type of adult stem cell, are easy to isolate, culture, and manipulate in ex vivo culture. These cells have great plasticity and the potential for therapeutic applications, but their properties are poorly understood. MSCs can be found in bone marrow and in many other tissues, and these cells are generally identified through a combination of poorly defined physical, phenotypic, and functional properties; consequently, multiple names have been given to these cell populations. Murine MSCs have been directly applied to a wide range of murine models of diseases, where they can act as therapeutic agents per se, or as vehicles for the delivery of therapeutic genes. In addition to their systemic engraftment capabilities, MSCs show great potential for the replacement of damaged tissues such as bone, cartilage, tendon, and ligament. Their pharmacological importance is related to four points: MSCs secrete biologically important molecules, express specific receptors, can be genetically manipulated, and are susceptible to molecules that modify their natural behavior. Due to their low frequency and the lack of knowledge on cell surface markers and their location of origin, most information concerning MSCs is derived from in vitro studies. The search for the identity of the mesenchymal stem cell has depended mainly on three culture systems: the CFU-F assay, the analysis of bone marrow stroma, and the cultivation of mesenchymal stem cell lines. Other cell populations, more or less related to the MSC, have also been described. Isolation and culture conditions used to expand these cells rely on the ability of MSCs, although variable, to adhere to plastic surfaces. Whether these conditions selectively favor the expansion of different bone marrow precursors or cause similar cell populations to acquire different phenotypes is not clear. The cell populations could also represent different points of a hierarchy or a continuum of differentiation. These issues reinforce the urgent need for a more comprehensive view of the mesenchymal stem cell identity and characteristics. PMID- 16370332 TI - Neovascularization and cardiac repair by bone marrow-derived stem cells. AB - Postinfarction congestive heart failure with impaired systolic left ventricular function is a loss of cardiomyocyte disease. Adult stem or progenitor cells from the bone marrow and the peripheral blood have been experimentally shown to differentiate towards endothelial cells and cardiomyocytes under the appropriate conditions. The use of autologous adult stem cells for neovascularization and cardiac regeneration is a promising concept and has shown benefit in pilot clinical trails enrolling postinfarction patients with coronary artery disease. Cell therapy may act through differentiation into and thus replacement of cardiomyocytes and/or neovascularization, the formation of new vessels in the adult organism. Moreover, the release of factors acting in a paracrine manner may contribute to neovascularization and scar remodelling. In this review, the experimental data regarding neovascularization and cardiomyocyte formation from adult stem/progenitor cells are discussed. PMID- 16370333 TI - The potential use of myogenic stem cells in regenerative medicine. AB - More than a century after the initial description of muscular dystrophy, no curative treatment is currently available. To date, clinical trials with myogenic stem cell transplantation have met with only modest success. There are multiple factors behind these failures, yet they provide powerful insights for improvement. In this chapter, we review the different myogenic stem cell populations that have been reported to be potential vectors for the treatment of myopathies in a context of regenerative medicine. PMID- 16370334 TI - Neural stem cells: on where they hide, in which disguise, and how we may lure them out. AB - In contrast to the haematopoietic system in which each cell type is subject to constant turnover, thus endowing this system with the permanent ability to reconstitute itself, the nervous system has long been known as an organ devoid of spontaneous cellular reconstitution. Yet the discovery that certain regions of the mammalian central nervous system do sustain neurogenesis throughout life, together with the fact that cells can be isolated from the adult brain that generate neurons in vitro, has led to the idea that the nervous tissue harbours neural stem cells. The term "neural stem cell" has now become associated with enormous expectations for curing diseases of the nervous system. Yet many of the biological fundamentals of neural stem cells need to be revealed before these expectations can be properly judged or even fulfilled. This begins with the question of whether the neural stem cell corresponds to a real entity or rather represents an in vitro dedifferentiation phenomenon. In this chapter we attempt to give an overview of our current knowledge of the biology of the presumable adult neural stem cell. This is followed by a comparative assessment of the possibilities of using adult neural stem cells and embryonic stem cells for therapeutic approaches in the context of neurodegenerative diseases. Finally, we will look at the "evil side" of stemness by discussing the evidence that brain cancers may originate from cells with stem cell-like properties. PMID- 16370335 TI - Cell transplantation for patients with Parkinson's disease. AB - This chapter focuses on cell replacement therapies in Parkinson's disease (PD) and describes experimental data leading to clinical trials, as well as the methodology and efficacy of grafts of human embryonic nigral tissue in patients with PD. It also highlights some of the clinical problems the procedure presents and considers future alternative sources of donor tissue, including various forms of stem cells. PMID- 16370336 TI - Postmodern biology: (adult) (stem) cells are plastic, stochastic, complex, and uncertain. AB - This chapter will discuss recent findings regarding cell plasticity and stem cell behavior, focusing on ways in which experimental design, observer interference, and inherent stochasticity and complexity are serving to create a new, postmodern biology. The chapter will summarize: (a) the four recognized pathways whereby cell plasticity occurs physiologically; (b) recent findings regarding unexpected epigenetic reversibility of gene restrictions that provide the mechanistic core of plasticity; (c) current evidence for the stochastic nature of gene expression and, therefore, of cell fate decisions. It will be noted that stochastic, however, does not imply completely random; rather, constrained randomness, intermediate between rigid determinism and complete disorder is what is usually seen experimentally. Possible sources of such constrained disorder, from a biomolecular point of view, will be discussed. The chapter will conclude with discussions of how these findings contribute to a Complexity Theory formulation of the body as self-organizing emergence of interacting biomolecules and the implications of such concepts for design and interpretation of experimental results (i.e., a cellular version of Heisenbergian uncertainty). PMID- 16370337 TI - [Sensory nerve anesthesia with 4% liposomal lidocaine cream in healthy subjects by use of the neuroselective current perception threshold]. AB - BACKGROUND: ELA-Max, an external liposomal preparation of lidocaine (4%) available without prescription in United States, has a recommended application time of 15 to 45 min without occlusive dressing, and the side effect has been reported to be rare. To investigate external anesthetic preparations to reduce neuropathic pain, we evaluated sensory nerve anesthesia by ELA-Max in comparison with placebo and other lidocaine topical preparations by double blind method. METHODS: We measured the neuroselective current perception threshold (N-CPT) 30 min after the application of ELA-Max on 30 healthy Japanese volunteers. Evaluation of anesthetic effect was performed by change in N-CPT rate before and after the application of 0.025 g x cm(-2) to the skin where stimulation was performed. RESULTS: ELA-Max increased N-CPT ratio in 4 out of 9 subjects stimulated at 5 Hz. However these CPT values were within normal ranges, and the change in N-CPT ratio was not statistically significant throughout all stimulation frequencies. CONCLUSIONS: The application of 0.025 g x cm(-2) ELA-Max for 30 min was not effective to induce therapeutic level of anesthesia. Extended application times as well as occlusive dressing may be needed for this preparation to be used clinically. PMID- 16370338 TI - [Anesthetic management of a cesarean section in a patient with schizophrenia receiving a large amount of antipsychotic agents]. AB - We experienced perioperative management of a patient with schizophrenia who had been taking a large amount of antipsychotic agents until the cesarean section. A 31-year-old woman in 38th week gestation with schizophrenia underwent a cesarean section. Anesthesia was maintained with light plane of general anesthesia combined with epidural anesthesia. Perioperatively, the mother experienced no complications and the baby only developed slight generalized tremor and hypotonus for a short period after delivery. PMID- 16370339 TI - [Brain protection with mild hypothermia during carotid artery clamping]. AB - A 71-year-old man was scheduled for removal of a Kirchner wire malpositioned in the mediastinum, which had been placed for fixing the fractured right clavicle five months before. Anesthesia was induced and maintained with propofol, fentanyl and vecuronium. The wire was found to be penetrating the brachiocephalic artery after sternotomy. An emergency angiography performed in the operating room showed that Willis arterial circle was sufficiently developed for clamping the brachiocephalic artery. The wire was removed under clamping the brachiocephalic artery for 9 minutes, but massive bleeding from the left common carotid artery continued, then the left common carotid artery was clamped and injured region was resected and reconstructed for 68 minutes. The body temperature was reduced to 32.5-33 degree with a cooling water mattress for brain protection and prostaglandin E1 was infused for vasodilation during hypothermia. Monitoring with somatosensory evoked potential was added during anesthesia. The surgery was performed uneventfully and the patient showed no neurological sequelae postoperatively. PMID- 16370340 TI - [Anesthetic management of a patient undergoing PTA stent placement for right common carotid artery stenosis]. AB - A 60-year-old man underwent percutaneous transluminal angioplasty (PTA) stent replacement of the right common carotid artery. Preoperative angiogram revealed bilateral vertebral artery occlusion and 50% stenosis in contralateral internal carotid artery. Anesthesia was induced and maintained with fentanyl and propofol by TCI. rSO2, BIS and EEG were monitored. Moderate hypothermia (33-35 degrees C) was induced by concomitant use of milrinone. Edaravone, a novel free radical scavenger, and Sendai cocktail were administered before interruption of carotid flow. During 5 minutes of test occlusion by balloon, right rSO2 decreased from 61% to 49% and EEG showed slow waves with decreased amplitude. Therefore we decided to perform PTA and stenting separately. Right rSO2 decreased from 62% to 48% during PTA (6 min occlusion), while rSO2 decreased from 66% to 50% during stenting (7.5 min occlusion). EEG also showed the similar changes as observed during test occlusion. After the procedures, rSO2 and EEG recovered in a short time. Postoperative angiogram showed an improvement of carotid artery stenosis and intracranial vessels showed no branch occlusion. Patient was maintained hypothermic (35 degrees C) for 2 days after surgery. He recovered without additional neurological complications. We found that rSO2 was a useful, real-time and non-invasive method for evaluation of cerebral ischemia in our patient. PMID- 16370341 TI - [Anesthetic management in a patient with Goldenhar's syndrome using a perilaryngeal airway and a videolaryngoscope]. AB - A 5-year-old girl with Goldenhar's syndrome was scheduled for reconstruction of her left thumb. Because her preanesthetic X-p had revealed airway abnormality, difficult intubation was anticipated. After induction of anesthesia with thiamylal, the lungs were ventilated easily via a facemask. A perilaryngeal airway (PLA) was inserted after obtaining adequate depth of anesthesia with sevoflurane. Anesthesia was maintained with sevoflurane (2%) plus nitrous oxide (66%) under spontaneous ventilation. The operation was finished uneventfully. However, a re-operation was scheduled for postoperative hemorrhage on that day. In consideration of potential regurgitation of gastric content, endotracheal intubation was scheduled. Her vocal cord was confirmed with a videolaryngoscope, and she was intubated successfully. In conclusion, a PLA and a videolaryngoscope are useful for airway management of patients with Goldenhar's syndrome for whom difficulty in intubation is anticipated. PMID- 16370342 TI - [General anesthesia for diaphragmatic herniorrhaphy in a patient with myasthenia gravis and severe gibbus]. AB - We experienced a case of diaphragmatic herniorrhaphy complicated with myasthenia gravis (MG) and gibbus. A 66-year-old woman was scheduled for diaphragmatic herniorrhaphy. She had a 10-year history of MG graded Osserman classification IIA. She had received prednisolone and pyridostigmine. After epidural catheter insertion into T 6-7 interspinal space, anesthesia was induced with sevoflurane 5% and maintained with sevoflurane 1%, nitrous oxide 3 l x min(-1) in oxygen 3 l x min(-1) and epidural block using 1% mepivacaine. Surgery was completed in 75 minutes. She was extubated without any complications 45 minutes after the end of surgery. Using muscle relaxant in this case with MG and gibbus might place her under artificial ventilation after surgery. For diaphragmatic herniorrhaphy adequate muscle relaxation is necessary. Thus, we used sevoflurane and epidural block. We successfully anesthetized a patient for diaphragmatic herniorrhaphy complicated with myasthenia gravis and gibbus by sevoflurane and epidural block. PMID- 16370343 TI - [A case of stagnation of a large amount of irrigation fluid in abdominal cavity during hysteroscopic myomectomy]. AB - A 30-year-old woman was scheduled to undergo hysteroscopic myomectomy for uterine myoma. Spinal anesthesia was performed with 3 ml of 0.5% bupivacaine and she was sedated with intravenous propofol during the surgery. After the surgical procedure, propofol infusion was stopped. The patient soon awoke and vomited frequently and repeatedly. Her serum sodium concentration was as low as 129 mEq x l(-1) and fluid overload (water intoxication) was suspected. Because the abdominal X-P revealed that two parts of the surgical instrument remained in her uterine cavity at that time, hysteroscopic procedure was restarted. General anesthesia was induced with propofol and maintained with nitrous oxide and sevoflurane. During the procedure, uterus was accidentally perforated and one of the parts moved into the abdominal cavity. When the laparotomy started to fix uterine perforation, the abdomen of the patient became remarkably distended. During the laparotomy, it was revealed that 4800 ml of irrigation fluid had been accumulated in the abdominal cavity. Although hysteroscopic surgery is generally considered non-invasive, two major complications (fluid overload and uterine perforation) occurred in this case. This case suggests that anesthetists must remain alert for the signs of these complications during hysteroscopic surgery. PMID- 16370344 TI - [How do you intubate the trachea?]. AB - We examined whether there was a difference in the description of the tracheal intubation technique among the Japanese textbooks of anesthesiology. We examined Japanese textbooks of anesthesiology stored in a university medical library. We found a considerable difference in the description concerning the technique of the tracheal intubation. To achieve the better compulsory clinical training, we think it is necessary to make a certain criterion for the tracheal intubation. PMID- 16370345 TI - [Outsourcing of anesthesia support system]. AB - BACKGROUND: Recently, the deficiency in anesthesiologist's man power became a social problem in Japan. Therefore, we tried outsourcing of anesthesia support system. METHODS: The questionnaire was sent to anesthesiologists after an anesthesia support system had started. RESULTS: All anesthesiologists answered reduction in the burden of work. CONCLUSIONS: Anesthesia support system outsourcing is effective in reducing the workload of anesthesiologists. PMID- 16370346 TI - [Perioperative evaluation of a kit for simplified measurement of blood theophylline concentration (Accumeter Theophylline)]. AB - BACKGROUND: We investigated the perioperative usefulness of a kit for simplified measurement of blood theophylline concentration (BTC), Accumeter theophylline (Nikken Chemicals Co.) in asthmatic patients. METHODS: Study 1) BTCs were measured by Accumeter at the beginning of surgery in asthmatic patients who had been medicated with oral theophylline. Study 2) BTCs were corrected during anesthesia in asthmatic patients who showed lower BTCs (<8 microg x g ml(-1)) at the beginning of surgery. Study 3) Theophylline was administered to asthmatic patients who had not been medicated with theophylline, and calculated and measured BTCs were compared. RESULTS: Study 1) Measured BTC at the begin- ning of surgery was 8.2 +/- 3.1 microg x ml(-1) (mean +/- SD), and 14 of 31 patients (45%) showed lower BTC. Patients who smoked showed a significantly lower BTC than that of non-smoking patients (P=0.04). Study 2) Five of 10 patients showed lower BTC at the beginning of surgery, but BTC had increased significantly at the end of surgery after i.v. administration of theophylline. Study 3) There was no significant linear correlation between calculated and measure BTCs after i.v. administration of theophylline (R2=0.25, n=10). Although the mean was near zero (0.05) by limits of agreement, 2 SD was large (2.53). There was no patient who showed wheezing during the study. CONCLUSIONS: It is important to monitor BTCs perioperatively in asthmatic patients, and Accumeter theophylline is a simple and useful device for management of BTC. PMID- 16370347 TI - [Anesthesia management may alter long-term outcome-review of recent reports]. AB - Perioperative management including anesthesia may alter long-term outcome of surgical patients. We have reviewed abstracts for meeting and articles published recently concerning effects of anesthetic depth, volatile anesthetic preconditioning, beta-blockers, alpha-agonists, statins, and glucose control on long-term outcome. Although research in this area has just been started, we, anesthesiologists should recognize its importance. PMID- 16370348 TI - [Considerations when conducting study of semen indices in workplaces]. AB - The possible adverse impact of environmental chemical exposure on semen quality is a public health concern. However, an epidemiological study targeting the general population is not easy. An alternative approach is to study workers exposed to higher doses of the concerned chemicals, whereas there have been only a very limited number of such studies conducted in Japan to date. This review, on the basis of the author's experience thus far, attempts to shed light on considerations when planning possible semen studies in workplaces in Japan. They include study design, informed consent of both the employer and participating employees, means of increasing participation ratio, control of nonexposure factors such as abstinence period and medical history, standardization in measuring semen indices, interpretation of study results, and necessary precautions when reporting a low semen quality in the exposed population. PMID- 16370349 TI - [Investigation of toxicity through human cases and animal experiments--study of bromopropanes]. AB - In 1995, workers in a Korean electronic factory exhibited oligospermia or amenorrhea. We investigated the toxicity of 2-bromopropane used as an alternative to chlorofluorocarbons in the factory in animal experiments, and clarified that exposure to 2-bromopropane depletes spermatogenic cells in male rats and oocytes in female rats. Subsequently, we investigated the neurotoxicity of 2-bromopropane on the basis of the reported neuropathy in the Korean workers exposed to 2 bromopropane. For comparison, we employed 1-bromopropane, which is now used as a new alternative to chlorofluorocarbons. The results showed that 1-bromopropane is more neurotoxic than 2-bromopropane, causing harm to reproductive organs by inhibiting spermiation in the testis and impairing follicular development in the ovary. Shortly after the initial investigation of 1-bromopropane in animals, human cases were reported in the United States. Neurologic abnormalities in Chinese workers exposed to 1-bromopropane were also reported, such as the elongation of distal latency and lowered sense of vibration in the lower limbs. Thus, these serial studies revealed that 1-bromopropane is neurotoxic, but its dose-response relationship in humans remains unknown. In the investigation of 2 bromopropane toxicity, initial animal studies were designed on the basis of human studies, while in research on 1-bromopropane, animal studies preceded human studies and contributed to the prediction of toxicity in humans. However, the use of animal model is limited in its ability to predict the toxicity of chemicals introduced depending on species differences. Further studies should focus more on the differences and commonality between animals and humans in response to toxic agents. PMID- 16370350 TI - [Significance of laboratory studies of neurobehavioral and developmental toxicities--transgenerational effects of styrene exposure]. AB - Recently, health problems caused by environmental chemical substances present in daily life have been increasing, particularly developmental toxicities, the effects of which often become apparent only after a long developmental period. It is difficult to determine adverse transgenerational effects. Therefore, we must recognize important indices to measure neurobehavioral and developmental effects. The assessment of such effects indices, is difficult and few neurobehavioral data are available compared to teratological data. Thus, through studies using laboratory animals, analyses of the mechanisms, exposure periods, doses and neurobehavioral effects are necessary. We have reviewed studies of the volatile monomer of the organic solvent styrene with regard to reproductive and developmental toxicities in laboratory animals. The styrene monomer crosses through the placenta; however, fetal excretion is less than maternal excretion. Therefore, it seems that a dose of styrene that does not affect the mother may be toxic for the fetus. We reported that exposure to low-dose styrene results in physical and neurobehavioral developmental delays, as well as decreases in enzyme activity and neurotransmitter secretion level. To prevent neurobehavioral toxicity, we need further studies to obtain precise data on chemical-biological interactions developmental toxicity and dose-response relationships. A key step towards effective prevention can also be obtained from studies in which animals are continuously exposed to chemicals for one or more generations. Thereafter, we must extrapolate developmental toxicity data from animals to humans. In this paper, we provide the information on developmental toxicity. PMID- 16370351 TI - [Trace elements and sperm parameters in semen of male partners of infertile couples]. AB - OBJECTIVE: The relationships between element concentrations and sperm parameters in semen samples were investigated. METHODS: Semen samples (n = 113) were donated voluntarily by male partners of infertile couples. The concentrations of fourteen elements (Na, K, P, Ca, Zn, Mg, Fe, Cu, Se, Mn, Sn, Co, Ni, and Cd) in semen were determined by atomic absorption spectrometry, fluorometry, or colorimetry. Element concentrations in seminal plasma and in sperm were also measured. RESULTS: Element concentrations in semen were in the order Na > P, K > Ca > Zn > Mg > > Fe> Cu, Se > Mn > Sn, Ni, Co, Cd. When the samples were divided into two groups in terms of sperm concentration and number, the Se concentration in semen with normal parameter values (sperm concentration > or = 20 x 10(6) and sperm number > or = 40 x 10(6)), 99.4 +/- 37.4 ng/ml, was higher than that in semen with abnormal parameter values (sperm concentration < or = 20 x 10(6) and/or sperm number < or = 40 x 10(6)), 72.1 +/- 33.9 ng/ml (p < 0.001). A clearer positive correlation between the Se concentration and the sperm concentration was observed in the sperm portion (r = 0.853, p < 0.001) than in semen (r = 0.512, p < 0.001) and seminal plasma (r = 0.292, p = 0.003). Statistically significant correlations were also observed between the concentration of Se, P, Zn, Cu, Fe, or Mn in semen, the sperm portion or seminal plasma and the sperm concentration, semen volume or abnormal morphology, although correlation coefficients were small. CONCLUSION: Among biologically essential elements in semen of infertile males, Se was a good indicator of sperm concentration; however, other trace elements did not indicate clear relationships between their concentrations and sperm parameters. PMID- 16370352 TI - [Hyperlipidemia: complex pathophysiology caused by multiple genetic and environmental factors--in considering the approaches to preventive medicine]. AB - In Japan, over the last 40 years, the change in lifestyle, particularly the westernization of the diet, has led to increased frequency of "lifestyle-related disorders" such as, hyperlipidemia, diabetes mellitus, and atherosclerosis. Consequently, the morbidity and mortality due to coronary heart disease have increased, and the prevention of this disorder is now one of the major concerns when considering the quality of life of individuals and the public health policy. Hyperlipidemia, particularly hypercholesterolemia, is one of the major risk factors of atherosclerosis. Therefore, the clarification of the mechanisms of the development of hyperlipidemia is important in the consideration of the prevention and management of atherosclerotic disorders. Genetic and epidemiological studies have provided compelling evidence that genetic factors, environmental influences and the interaction between them all contribute to the development of this complex disorder. Various unexpected mechanisms of the development of hyperlipidemia have been elucidated by analyzing some hereditary disorders. Furthermore, the cloning of the genes involved in the maintenance of cholesterol and/or lipid homeostasis has made it possible to investigate the molecular mechanisms of the development of hyperlipidemia in detail. Elucidation of the genetic and environmental factors and detailed understanding of the molecular pathophysiology of hyperlipidemia are essential in the pursuit of evidence-based preventive medicine and health policy. PMID- 16370353 TI - [Relationship between the change of mental health and the improvement of lifestyle and physical health]. AB - OBJECTIVES: The purpose of this study was to clarify the relationship between the change of mental health and the improvement of lifestyle and physical health in the lifestyle intervention program. METHODS: The study was conducted using data from 126 persons (50 males and 76 females) who participated in the 6-month lifestyle intervention program. We used the result of the General Health Questionnaire (GHQ)-30 as the indicator of mental health, the number of steps, surveys concerning caloric intake, and sleeping hours as the indicator of lifestyle, and body weight, BMI, blood pressure, triglycerides, total cholesterol, LDL cholesterol, HDL cholesterol and HBA1c. The subjects were classified into four groups based on the change of mental health: Group-GG, change from good to good (n = 80); Group-PG, change from poor to good (n = 25); Group-PP, change from poor to poor (n = 13); and Group-GP, change from good to poor (n = 8). The changes between pre-intervention and post-intervention were compared for each group by the paired t-test, and among the 4 groups by ANCOVA. RESULTS: In Group-GG, the number of steps (p < 0.01), calorie intake (p < 0.05) and sleeping hours (p < 0.01) were significantly improved, triglyceride was significantly decreased (p < 0.01), and total and HDL cholesterol were significantly increased (p < 0.01). However there were no significant changes in Group-GP. CONCLUSION: The state and change of mental health were found to be significantly associated with the improvement of lifestyle and physical health. PMID- 16370354 TI - [Nosocomial outbreak of scabies in a psychiatric hospital--epidemiological assessment and prophylactic treatment with oral ivermectin]. AB - OBJECTIVES: To evaluate the effectiveness of mass treatment with ivermectin of scabies outbreak in institutional settings. To determine the factors, such as host susceptibility and scabetic exposure level associated with the onset of scabies. METHODS: The authors investigated a nosocomial scabies outbreak in a close psychiatric ward. The index case was a man with steroid-induced localized crusted scabies. Twenty-six patients were diagnosed with scabies, 4 of them had relapse of scabies, while no staff was infested. Despite frequent surveillance and treatment of symptomatic patients with 1% gamma-benzenehexachloride (gamma BHC: Lindane), new cases were observed. Thus, all 69 patients in the ward were treated with ivermectin (200 microg/kg) simultaneously on day 105 of the outbreak (the mass treatment). Patients who had scabies were compared with patients who had no scabies in terms of age, body weight, diabetes, physical functions, topical administration of corticosteroid, proximity to the index patient, and problematic behavior. RESULTS: The mass treatment was implemented without a significant adverse event. Although two patients developed symptoms of scabies after the mass treatment, no patient in the ward had been diagnosed with scabies since the 98th day of the treatment. Regarding factors associated with the scabies onset, the only statistically significant factor was proximity to the index patient with crusted scabies. CONCLUSIONS: Oral ivermectin was safe and effective for controlling scabies in institutional settings. The exposure level to scabetic mites was more important than host susceptibility in determining the risk of scabies onset. PMID- 16370355 TI - [GGAs: structure and function of a novel family of clathrin adaptors involved in membrane trafficking between the TGN and endosomes]. PMID- 16370356 TI - [Biosynthesis and function of sulfated carbohydrate chains]. PMID- 16370357 TI - [Mechanism of chromosome segregation in meiosis]. PMID- 16370358 TI - [Mechanism of polarized distribution of intracellular components by class V myosin in the budding yeast]. PMID- 16370359 TI - [Male germline stem cells: stem cells for germline modification and regenerative medicine]. PMID- 16370360 TI - [Role of angiopoietin-like protein (Angptl)]. PMID- 16370361 TI - [Proteins involved in longevity]. PMID- 16370362 TI - [Sperm acrosome reaction in sea urchin at the intersection of glycobiology and evolutional biology]. PMID- 16370363 TI - [Molecular mechanism of axon guidance mediated by phosphorylation of CRMP2]. PMID- 16370364 TI - [Endogenous DNA damage in carcinogenesis]. PMID- 16370365 TI - [Physiological functions of phospholipase Cdelta]. PMID- 16370366 TI - Coming clean. PMID- 16370367 TI - Small molecules thwart crash and burn. PMID- 16370368 TI - Toward small-molecule agonists of TNF receptors. PMID- 16370369 TI - NO link between calorie restriction and mitochondria. PMID- 16370370 TI - Lactate transportation is required for lymphocyte activation. PMID- 16370371 TI - Photochemical tools for remote control of ion channels in excitable cells. AB - Various strategies have been developed recently for imparting light sensitivity onto normally insensitive cells. These include expression of natural photosensitive proteins, photolysis of caged agonists of native cell surface receptors and photoswitching of isomerizable tethered ligands that act on specially engineered ion channels and receptor targets. The development of chemical tools for optically stimulating or inhibiting signaling proteins has particular relevance for the nervous system, where precise, noninvasive control is an experimental and medical necessity. PMID- 16370372 TI - Monocarboxylate transporter MCT1 is a target for immunosuppression. AB - Current immunosuppressive therapies act on T lymphocytes by modulation of cytokine production, modulation of signaling pathways or by inhibition of the enzymes of nucleotide biosynthesis. We have identified a previously unknown series of immunomodulatory compounds that potently inhibit human and rat T lymphocyte proliferation in vitro and in vivo in immune-mediated animal models of disease, acting by a novel mechanism. Here we identify the target of these compounds, the monocarboxylate transporter MCT1 (SLC16A1), using a strategy of photoaffinity labeling and proteomic characterization. We show that inhibition of MCT1 during T lymphocyte activation results in selective and profound inhibition of the extremely rapid phase of T cell division essential for an effective immune response. MCT1 activity, however, is not required for many stages of lymphocyte activation, such as cytokine production, or for most normal physiological functions. By pursuing a chemistry-led target identification strategy, we have discovered that MCT1 is a previously unknown target for immunosuppressive therapy and have uncovered an unsuspected role for MCT1 in immune biology. PMID- 16370373 TI - C3-symmetric peptide scaffolds are functional mimetics of trimeric CD40L. AB - Interaction between CD40, a member of the tumor necrosis factor receptor (TNFR) superfamily, and its ligand CD40L, a 39-kDa glycoprotein, is essential for the development of humoral and cellular immune responses. Selective blockade or activation of this pathway provides the ground for the development of new treatments against immunologically based diseases and malignancies. Like other members of the TNF superfamily, CD40L monomers self-assemble around a threefold symmetry axis to form noncovalent homotrimers that can each bind three receptor molecules. Here, we report on the structure-based design of small synthetic molecules with C3 symmetry that can mimic CD40L homotrimers. These molecules interact with CD40, compete with the binding of CD40L to CD40, and reproduce, to a certain extent, the functional properties of the much larger homotrimeric soluble CD40L. Architectures based on rigid C3-symmetric cores may thus represent a general approach to mimicking homotrimers of the TNF superfamily. PMID- 16370374 TI - Analysis of drug-induced effect patterns to link structure and side effects of medicines. AB - The high failure rate of experimental medicines in clinical trials accentuates inefficiencies of current drug discovery processes caused by a lack of tools for translating the information exchange between protein and organ system networks. Recently, we reported that biological activity spectra (biospectra), derived from in vitro protein binding assays, provide a mechanism for assessing a molecule's capacity to modulate the function of protein-network components. Herein we describe the translation of adverse effect data derived from 1,045 prescription drug labels into effect spectra and show their utility for diagnosing drug induced effects of medicines. In addition, notwithstanding the limitation imposed by the quality of drug label information, we show that biospectrum analysis, in concert with effect spectrum analysis, provides an alignment between preclinical and clinical drug-induced effects. The identification of this alignment provides a mechanism for forecasting clinical effect profiles of medicines. PMID- 16370375 TI - Gene array analysis to determine the components of neuropathic pain signaling. AB - The molecular modification of the pain pathway represents one of the major mechanisms underlying neuropathic pain. Recently, gene array studies have been carried out to identify the genes that are regulated at the spinal cord level after peripheral nerve injury. These studies demonstrate that peripheral nerve injury causes marked changes in gene expression in both the dorsal root ganglion (DRG) and the dorsal spinal cord. The markedly regulated molecules include, for example, neuropeptides, receptors, ion channels, signal transduction molecules and synaptic vesicle proteins. Upregulation of the Ca2+ channel alpha2/delta1 subunit, gamma-aminobutyric acid A receptor alpha5 subunit, Na+ channels and nicotinic acetylcholine receptors in the DRG and dorsal spinal cord indicates their potential roles in neuropathic pain control. PMID- 16370376 TI - Functional proteomic screens in therapeutic protein drug discovery. AB - Biopharmaceuticals, mainly protein-based therapeutics, are rapidly being developed for several disease indications. However, most biopharmaceuticals are 'me-too' drugs or are being developed against the same handful of targets. Thus, the potential of biopharmaceuticals is not being fully exploited. The bottleneck is still the lack of validated targets and the means of implementing the appropriate target validation technology. Functional proteomic screens provide a rapid route to accelerate the discovery and development of the next generation of biopharmaceuticals. PMID- 16370377 TI - Genomic studies in stem cell systems. AB - Stem cell research holds promise for future regenerative medicine. In recent years, much effort has been devoted to our understanding of stem cell self renewal and differentiation at the molecular level. Important studies that have applied various genomic approaches, including genomics, functional genomics and proteomics, to gain molecular insights into stem cell development, are reviewed. PMID- 16370378 TI - Stem cell pharmacogenomics: a reality check on stem cell therapy. AB - Widespread applications of stem cell-based therapies in the clinic are being reported, yet there is a surprising lack of resolution of the factors that lead to failure of such therapies on a long-term basis. While classical pharmacogenomics aids the prediction of drug responses in an individual based on genetic variation and pharmacological responses, stem cell therapy involves an additional dimension of host-donor cell interactions and adaptability. We propose the development of concise guidelines based on pharmacogenetic and donor/host related cellular factors studied more extensively in larger data sets, and used in prospective studies to individualize stem cell therapy. PMID- 16370379 TI - A proteomic approach to assess intraneuronal inclusions associated with neurodegenerative disorders. AB - In neuroscience, proteomic technology is being used to discover the chemical features of neurodegenerative disorders, including Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis. Pathologically, one hallmark feature common to these diseases is the presence of proteinaceous inclusions within affected neurons. Proteomic assessment of diseased tissues and animal models reveals that the occurrence of these protein-rich aggregations may be due to perturbed functioning of a neuron-specific ubiquitin-recycling enzyme. PMID- 16370380 TI - Integration of environment and disease into 'omics' analysis. AB - Over the last decade, 'omics' technologies have been heralded as 'dream toolboxes' that will make revolutionary changes in disease prevention and treatment possible. However, these expectations have yet to be fulfilled. These promises were made and prospects predicted without full appreciation of the intrinsic complexity of common diseases and the significant contribution of environmental factors to disease risk and causation, which have hardly been considered in the omic approaches. The concept of gene-environment interaction, as well as the limitations, difficulties and urgent need to integrate environment into omics analysis are reviewed and discussed. PMID- 16370381 TI - Heart failure in the post-genomics era: gene-environment interactions. AB - Cardiovascular diseases remain the major cause of mortality and morbidity in the Western world. Heart failure is the most rapidly rising cardiovascular condition and is associated with high mortality. The etiology of heart failure is multiple, ranging from genetic cardiomyopathies to structural modifications to the heart following myocardial infarction or long-standing high blood pressure. Molecular system biology techniques (microarrays and proteomics) in combination with bioinformatics can now provide unique insights into the molecular mechanisms leading to heart failure. Both gene-gene and gene-environment interactions determine the specific phenotype and outcomes in this condition. The identification of these pathways also provides opportunities for the discovery of novel diagnostic and/or prognostic markers, and important therapeutic targets. How recent applications of genomics technologies depict a more complete portrait of molecular events in heart failure is summarized. PMID- 16370382 TI - Technology evaluation: ipilimumab, Medarex/Bristol-Myers Squibb. AB - Medarex and Bristol-Myers Squibb are developing ipilimumab, an immunostimulatory human antibody against cytotoxic T-lymphocyte antigen-4, for the potential combination or monotherapy treatment of melanoma, prostate, breast, renal and other cancers, as well as HIV infection. PMID- 16370383 TI - Technology evaluation: NeuroVax, Immune Response Corp. AB - The Immune Response Corp is developing NeuroVax, a combination vaccine of three T cell receptor peptides (BV5S2, BV6S5 and BV13S1) in incomplete Freund's adjuvant, as a potential treatment for multiple sclerosis. The vaccine is currently undergoing phase II clinical trials. PMID- 16370384 TI - Technology evaluation: denosumab, Amgen. AB - Amgen, as part of its program targeting the RANK/RANKL/ osteoprotegerin pathway, is developing denosumab, a fully human monoclonal antibody, delivered subcutaneously, targeting the receptor activator of nuclear factor-kappaB ligand, for the potential treatment of diseases associated with bone loss, such as osteoporosis and bone metastases. The antibody is currently undergoing phase III clinical trials. PMID- 16370385 TI - Apoptosis as the predominant tumor cell response to chemotherapy and irradiation: a case of TUNEL vision? PMID- 16370386 TI - Therapeutic strategies for the treatment of neuroblastoma. AB - Neuroblastoma, the most common extracranial solid tumor in children, has a highly heterogeneous clinical presentation and course. Current risk-based therapy is usually effective in patients who have intermediate risk features, however, intensive treatment of advanced neuroblastoma in children over two years of age is far from satisfactory. Current therapeutic approaches include the optimization of pharmacokinetic and pharmacodynamic properties of conventional agents, as well as the development of novel targeted drugs, such as signal transduction and angiogenesis inhibitors, apoptosis/differentiation stimulators and immunotherapeutics. This review provides an overview of current treatment options and future perspectives for the therapy and prevention of neuroblastoma. PMID- 16370387 TI - Potential role of the antiproliferative cytokine beta-galactoside binding protein in cancer therapy. AB - This review highlights the discovery of beta-galactoside binding protein (betaGBP), an antiproliferative cytokine, as a potent and selective anticancer agent. Unlike drugs designed to block receptor tyrosine kinases or specific control points along signaling pathways, betaGBP does not harm normal cells. As a physiological effector molecule, betaGBP can selectively induce death in cancer cells by enforcing its regulatory functions to which normal and cancer cells respond differently, and thus exploit the genetic and molecular deviations developed by cancer cells. PMID- 16370388 TI - Interleukin-2 for the treatment of melanoma. AB - Current therapeutic options for the treatment of advanced melanoma are inadequate. Chemotherapy with dacarbazine remains a 'gold standard' despite no evidence of improved survival or durable remissions. Interleukin (IL)-2 is an immunotherapeutic agent that, when administered in a high-dose bolus schedule, produces a small number of durable remissions in patients with metastatic melanoma, and on this basis it was approved for use in the US in 1998. In randomized clinical trials, IL-2, administered as a continuous infusion either alone or in combination with chemotherapy (biochemotherapy), has not improved response rates over chemotherapy alone. Low-dose IL-2, while less toxic and more convenient, produces low response rates and appears to be ineffective in metastatic melanoma. Newer, innovative approaches such as IL-2 gene therapy and strategies to ameliorate the toxicity of this agent are now being explored in clinical trials. PMID- 16370389 TI - Vaccination therapy with tumor-dendritic cell hybrids: a promising therapeutic approach? AB - Dendritic cells (DCs) are potent antigen-presenting cells currently being investigated as a tool for antitumor vaccination strategies. As an alternative to loading DCs with tumor antigen preparations, hybrid cells generated by fusing tumor cells with DCs have successfully been evaluated in preclinical cancer models. Hybrid cells express a large repertoire of tumor-associated antigens, high levels of major histocompatibility complex class I and II molecules, and adhesion/co-stimulatory molecules. Therefore, these cells possess properties of both parental cell types that are necessary for the induction of primary helper and cytotoxic T-cell responses. The results of early clinical trials suggest that hybrid cell vaccination is a safe and well-tolerated procedure capable of inducing T-cell responses. However, the few objective clinical responses observed indicate that further optimizations are required. PMID- 16370390 TI - Growth factor receptors: implications in tumor biology. AB - Growth factors are signaling molecules which bind to cell surface receptors and mediate a myriad of intracellular functions. Growth factor signaling is vital for growth and differentiation of cells under normal physiological conditions. However, aberrant signaling of these molecules via their receptors enables the cells to acquire abnormal characteristics most commonly observed in tumor cells. Tumor biology studies have revealed a central role for growth factor receptors in tumor progression. This review discusses the involvement of growth factor receptors in solid tumor formation and their value as potential anticancer drug targets. PMID- 16370392 TI - EndoTAG-1. MediGene. AB - MediGene is developing EndoTAG-1, a formulation of paclitaxel encapsulated in positively charged lipid-based complexes, for the potential treatment of cancer. Phase II clinical trials in patients with advanced pancreatic cancer are ongoing. PMID- 16370391 TI - Targeting the phosphatidylinositol-3 kinase/Akt pathway for the treatment of cancer. AB - Phosphatidylinositol-3 kinase (PI3K)/Akt is overactivated in a wide range of tumor types, and this triggers a cascade of responses, from cell growth and proliferation to survival, motility, epithelial-mesenchymal transition and angiogenesis. Therefore, this pathway presents an exciting target for molecular therapeutics. In addition, ectopic expression of PI3K or Akt, especially constitutively activated PI3K (p110alpha) or Akt, is sufficient to induce the oncogenic transformation of cells and tumor formation in transgenic mice, as well as the development of chemoresistance. Inhibition of PI3K/Akt signaling induces apoptosis and inhibits the growth of tumor cells that have elevated Akt levels. The dependence of certain tumors on PI3K/Akt signaling for survival and growth has wide implications for cancer therapy, offering the potential for preferential tumor cell killing. In the past few years, a number of inhibitors of the Akt pathway have been identified by combinatorial chemistry, high-throughput and virtual screening, or traditional medicinal chemistry. This review focuses on ongoing translational efforts to therapeutically target the PI3K/Akt pathway. PMID- 16370393 TI - MDX-060. Medarex. AB - Medarex is developing the antibody MDX-060 for the potential treatment of CD30+ lymphomas, such as Hodgkin's disease and anaplastic large cell lymphomas. Phase I/II clinical trials were underway by December 2002. PMID- 16370394 TI - IGN-311. Igeneon. AB - Igeneon, under license from Protein Design Labs, is developing IGN-311, an intravenous humanized monoclonal antibody against the Lewis Y carbohydrate antigen, as a potential agent for the passive immunotherapy of cancer, particularly epithelial tumors affecting breast, colon, gastric and pancreatic tissues. A phase I/II trial of IGN-311 was initiated in July 2005. PMID- 16370395 TI - Focused ion beam characterization of plasma-assisted deposition on polymer films at the nanoscale. AB - In this paper, a novel technique is presented for the characterization at the nanoscale of plasma-assisted deposit on polyethylene-terephthalate (PET) polymer films. In previous studies, some microcharacterization and morphology analyses of plasma-assisted deposition were performed by atomic force microscopy (AFM). In the work presented here, we analysed the thickness and homogeneity of plasma assisted deposits by focused ion beam (FIB). This technique with 5-7 nm resolution requires no sample preparation and relies on a sequence of operations on a relatively fast time scale, so that it is easy to make thorough investigations of the sample. We performed electron and ion imaging of the surface of the material, and a subsequent ionic cutting allowed the study of the morphology of the same sample. We developed a novel approach to the edge detection techniques (EDT) in images for a fast evaluation and monitoring of the deposited layer. PMID- 16370396 TI - An engineered microenvironment for multidimensional microscopy of live cells. AB - Multidimensional imaging (MD) of live cells is gaining importance in biomedical research as the commercial availability of confocal, nonlinear optical microscopes, environmental chambers, and specific fluorescence probes grows. One crucial aspect of the MD live cell imaging involves the proper immobilization of cells, which refers to the rapid and sufficient immobilization of cells on the microscope stage, neither disrupting the cellular structure and functions nor affecting the optical properties of the cells and the environments. Conventional cell immobilization methods glue the anchoring cells to coated surfaces, but such methods require centrifugation or extended incubation and are not suitable for cells in suspension. Most of the current three-dimensional (3-D) gels either exhibit unsatisfactory optical properties or have adverse effects on cell functions in culture. Recently, an engineered 3-D microcapsule has been developed that involves the complex coacervation of a positively charged collagen and a negatively charged polymer of 2-hydroxyethyl methacrylate--methacrylic acid- methyl methacrylate (HEMA-MMA-MAA). Hence, confocal imaging of live cells in this engineered 3-D microenvironment was investigated for its optical properties and cellular function compatibility. We report here that this microenvironment facilitates efficient cell immobilization, exhibits good optical properties, and can preserve cellular structures and functions, which will be useful in MD imaging of live cells for various applications. PMID- 16370397 TI - An experimental model of beam broadening in the variable pressure scanning electron microscope. AB - In the variable pressure scanning electron microscope (VP-SEM) the incident electrons pass through a gaseous environment and the beam is scattered by these interactions. We show here that the experimental intensity profile of the scattered beam can be described as Gaussian in form to a high level of accuracy. This provides a rapid means of accounting for the effects of beam scatter in imaging and microanalysis because the standard deviation of the Gaussian is a simple function of parameters such as working distance, beam energy, gas type and pressure. PMID- 16370398 TI - Measuring the angular dependent energy distribution of backscattered electrons at variable geometry. AB - An aluminium semisphere system with 120 points of entry and eight detection areas, assembled on a meridian covering 0.0026 steradian each, was put over a solid bulk sample (e.g., aluminium), which was mounted in the eucentric point so that the incident electron beam could be varied by a polar rotation of the sphere in steps of 11.25 degrees. The complete angular distribution of the backscattered electrons became available by a rotation in steps of 11.25 degrees azimuthally. For this particular setup, the signals from the detection areas as well as the signal from the rest of the semisphere were amplified by operational amplifiers (Burr-Brown OPA128LM). However the signal of the semisphere was not available at that time. Specimen current measurements made the total amount of electrons accessible, providing a possibility for normalization of the results and comparison with total backscattering coefficients. By use of counter voltage variable up to 10 kV inside the detection assembly, it was possible to measure an energy resolution of the backscattered electrons for each detection area at the same time. Details of the construction and calibration procedures, possible errors, and sources of systematic deviations as well as first test results are discussed. PMID- 16370399 TI - Scanning electron microscopy of some endophytic streptomycetes in snakevine- Kennedia nigricans. AB - Soils of all types and locations have generally served as the major sources of streptomycetous bacteria. These organisms are the source of nearly 80% of the world's antibiotics. Now, it is realized that Streptomyces spp. (within the group of prokaryotic filamentous bacteria known as actinomycetes) can exist as endophytes within the interstices of some higher plants. While it is sometimes possible to isolate one or two different streptomycetes from certain plants, most plants are free of these organisms. However, the snakevine (Kennedia nigricans) of the Northern Territory of Australia has yielded at least 39 different endophytic actinomycetes (95% of them being Streptomyces spp.) Most of these isolates possessed no detectable antibiotic properties, while at least seven had antibacterial and antifungal activities. Examination of eight selected cultures by scanning electron microscopy (SEM) as well as environmental scanning electron microcopy (FEI ESEM FEG) (FEI Company, Hillsobro, Ore., USA) revealed unusual patterns, structures, and features of the spores and hyphae of these microorganisms. For instance, as revealed by ESEM FEG for the first time, it has become obvious that extremely fine hair-like structures (average 25-49 nm with gold-coated specimens) exist on the spores and hyphae of some endophytic streptomycetes. The biological purpose of these hair-like protrusions is unknown. Both SEM and ESEM FEG can be effectively used as tools in identification and elucidation of the biology of these organisms. In addition, unusual colony morphology, observed with the unaided eye can very easily be used to distinguish some of these isolates since characteristic donut and pseudo-horn shaped colonies appeared in culture. PMID- 16370400 TI - Case report: posterior reversible encephalopathy syndrome. PMID- 16370402 TI - Integrating genetic counseling into family medicine. PMID- 16370401 TI - The postpartum visit: is six weeks too late? PMID- 16370403 TI - Management of Staphylococcus aureus infections. AB - Because of high incidence, morbidity, and antimicrobial resistance, Staphylococcus aureus infections are a growing concern for family physicians. Strains of S. aureus that are resistant to vancomycin are now recognized. Increasing incidence of unrecognized community-acquired methicillin-resistant S. aureus infections pose a high risk for morbidity and mortality. Although the incidence of complex S. aureus infections is rising, new antimicrobial agents, including daptomycin and linezolid, are available as treatment. S. aureus is a common pathogen in skin, soft-tissue, catheter-related, bone, joint, pulmonary, and central nervous system infections. S. aureus bacteremias are particularly problematic because of the high incidence of associated complicated infections, including infective endocarditis. Adherence to precautions recommended by the Centers for Disease Control and Prevention, especially handwashing, is suboptimal. PMID- 16370404 TI - Urinary tract infection in children. AB - Up to 7 percent of girls and 2 percent of boys will have a symptomatic, culture confirmed urinary tract infection by six years of age. Urinary tract infection may be suspected because of urinary symptoms in older children or because of fever, nonspecific symptoms, or failure to thrive in infants. Urine dipstick analysis is useful for ruling out urinary tract infections in cases with low clinical suspicion. However, urine culture is necessary for diagnosis of urinary tract infections in children if there is high clinical suspicion, cloudy urine, or if urine dipstick testing shows positive leukocyte esterase or nitrite activity. Despite current recommendations, routine imaging studies (e.g., renal ultrasonography, voiding cystourethrography, renal scans) do not appear to improve clinical outcomes in uncomplicated urinary tract infections. Oral antibiotics are as effective as parenteral therapy in randomized trials. The optimal duration of antibiotic therapy has not been established, but one-day therapies have been shown to be inferior to longer treatment courses. PMID- 16370405 TI - An approach to the postpartum office visit. AB - The postpartum period (typically the first six weeks after delivery) may underscore physical and emotional health issues in new mothers. A structured approach to the postpartum office visit ensures that relevant conditions and concerns are discussed and appropriately addressed. Common medical complications during this period include persistent postpartum bleeding, endometritis, urinary incontinence, and thyroid disorders. Breastfeeding education and behavioral counseling may increase breastfeeding continuance. Postpartum depression can cause significant morbidity for the mother and baby; a postnatal depression screening tool may assist in diagnosing depression-related conditions. Decreased libido can affect sexual functioning after a woman gives birth. Physicians should also discuss contraception with postpartum patients, even those who are breastfeeding. Progestin-only contraceptives are recommended for breastfeeding women. The lactational amenorrhea method may be a birth control option but requires strict criteria for effectiveness. PMID- 16370406 TI - Information from your family docter. Taking care of yourself after having a baby. PMID- 16370407 TI - Treatment of irritable bowel syndrome. AB - Irritable bowel syndrome affects 10 to 15 percent of the U.S. population to some degree. This condition is defined as abdominal pain and discomfort with altered bowel habits in the absence of any other mechanical, inflammatory, or biochemical explanation for these symptoms. Irritable bowel syndrome is more likely to affect women than men and is most common in patients 30 to 50 years of age. Symptoms are improved equally by diets supplemented with fiber or hydrolyzed guar gum, but more patients prefer hydrolyzed guar gum. Antispasmodic agents may be used as needed, but anticholinergic and other side effects limit their use in some patients. Loperamide is an option for treatment of moderately severe diarrhea. Antidepressants have been shown to relieve pain and may be effective in low doses. Trials using alosetron showed a clinically significant, although modest, gain over placebo, but it is indicated only for women with severe diarrhea predominant symptoms or for those in whom conventional treatment has failed. Tegaserod has an advantage over placebo in constipation-predominant irritable bowel syndrome; it is indicated for up to 12 weeks of treatment in women. However, postmarketing reports of severe diarrhea and ischemic colitis further limit its use. Herbal therapies such as peppermint oil also may be effective in the treatment of irritable bowel syndrome. Therapies should focus on specific gastrointestinal dysfunctions (e.g., constipation, diarrhea, pain), and medications only should be used when nonprescription remedies do not work or when symptoms are severe. PMID- 16370408 TI - Predicting the risk of recurrence after surgery for prostate cancer. PMID- 16370409 TI - Unilateral vision loss with a macular star. PMID- 16370410 TI - Hormone therapy for the prevention of chronic conditions in postmenopausal women. PMID- 16370411 TI - Rifaximin (Xifaxan) for traveler's diarrhea. PMID- 16370412 TI - Results of publicly-funded scientific research should be immediately available without cost to the public. For the proposition. PMID- 16370413 TI - Results of publicly-funded scientific research should be immediately available without cost to the public. Against the proposition. PMID- 16370414 TI - A three-dimensional reconstruction algorithm for an inverse-geometry volumetric CT system. AB - An inverse-geometry volumetric computed tomography (IGCT) system has been proposed capable of rapidly acquiring sufficient data to reconstruct a thick volume in one circular scan. The system uses a large-area scanned source opposite a smaller detector. The source and detector have the same extent in the axial, or slice, direction, thus providing sufficient volumetric sampling and avoiding cone beam artifacts. This paper describes a reconstruction algorithm for the IGCT system. The algorithm first rebins the acquired data into two-dimensional (2D) parallel-ray projections at multiple tilt and azimuthal angles, followed by a 3D filtered backprojection. The rebinning step is performed by gridding the data onto a Cartesian grid in a 4D projection space. We present a new method for correcting the gridding error caused by the finite and asymmetric sampling in the neighborhood of each output grid point in the projection space. The reconstruction algorithm was implemented and tested on simulated IGCT data. Results show that the gridding correction reduces the gridding errors to below one Hounsfield unit. With this correction, the reconstruction algorithm does not introduce significant artifacts or blurring when compared to images reconstructed from simulated 2D parallel-ray projections. We also present an investigation of the noise behavior of the method which verifies that the proposed reconstruction algorithm utilizes cross-plane rays as efficiently as in-plane rays and can provide noise comparable to an in-plane parallel-ray geometry for the same number of photons. Simulations of a resolution test pattern and the modulation transfer function demonstrate that the IGCT system, using the proposed algorithm, is capable of 0.4 mm isotropic resolution. The successful implementation of the reconstruction algorithm is an important step in establishing feasibility of the IGCT system. PMID- 16370415 TI - Simultaneous optimization of sequential IMRT plans. AB - Radiotherapy often comprises two phases, in which irradiation of a volume at risk for microscopic disease is followed by a sequential dose escalation to a smaller volume either at a higher risk for microscopic disease or containing only gross disease. This technique is difficult to implement with intensity modulated radiotherapy, as the tolerance doses of critical structures must be respected over the sum of the two plans. Techniques that include an integrated boost have been proposed to address this problem. However, clinical experience with such techniques is limited, and many clinicians are uncomfortable prescribing nonconventional fractionation schemes. To solve this problem, we developed an optimization technique that simultaneously generates sequential initial and boost IMRT plans. We have developed an optimization tool that uses a commercial treatment planning system (TPS) and a high level programming language for technical computing. The tool uses the TPS to calculate the dose deposition coefficients (DDCs) for optimization. The DDCs were imported into external software and the treatment ports duplicated to create the boost plan. The initial, boost, and tolerance doses were specified and used to construct cost functions. The initial and boost plans were optimized simultaneously using a gradient search technique. Following optimization, the fluence maps were exported to the TPS for dose calculation. Seven patients treated using sequential techniques were selected from our clinical database. The initial and boost plans used to treat these patients were developed independently of each other by dividing the tolerance doses proportionally between the initial and boost plans and then iteratively optimizing the plans until a summation that met the treatment goals was obtained. We used the simultaneous optimization technique to generate plans that met the original planning goals. The coverage of the initial and boost target volumes in the simultaneously optimized plans was equivalent to the independently optimized plans actually used for treatment. Tolerance doses of the critical structures were respected for the plan sum; however, the dose to critical structures for the individual initial and boost plans was different between the simultaneously optimized and the independently optimized plans. In conclusion, we have demonstrated a method for optimization of initial and boost plans that treat volume reductions using the same dose per fraction. The method is efficient, as it avoids the iterative approach necessitated by currently available TPSs, and is generalizable to more than two treatment phases. Comparison with clinical plans developed independently suggests that current manual techniques for planning sequential treatments may be suboptimal. PMID- 16370416 TI - Improving IMRT quality control efficiency using an amorphous silicon electronic portal imager. AB - An amorphous silicon electronic portal imaging device (EPID) has been investigated to determine its usefulness and efficiency for performing linear accelerator quality control checks specific to step and shoot intensity modulated radiation therapy (IMRT). Several dosimetric parameters were measured using the EPID: dose linearity and segment to segment reproducibility of low dose segments, and delivery accuracy of fractions of monitor units. Results were compared to ion chamber measurements. Low dose beam flatness and symmetry were tested by overlaying low dose beam profiles onto the profile from a stable high-dose exposure and visually checking for differences. Beam flatness and symmetry were also calculated and plotted against dose. Start-up reproducibility was tested by overlaying profiles from twenty successive two monitor unit segments. A method for checking the MLC leaf calibration was also tested, designed to be used on a daily or weekly basis, which consisted of summing the images from a series of matched fields. Daily images were coregistered with, then subtracted from, a reference image. A threshold image showing dose differences corresponding to > 0.5 mm positional errors was generated and the number of pixels with such dose differences used as numerical parameter to which a tolerance can be applied. The EPID was found to be a sensitive relative dosemeter, able to resolve dose differences of 0.01 cGy. However, at low absolute doses a reproducible dosimetric nonlinearity of up to 7% due to image lag/ghosting effects was measured. It was concluded that although the EPID is suitable to measure segment to segment reproducibility and fractional monitor unit delivery accuracy, it is still less useful than an ion chamber as a tool for dosimetric checks. The symmetry/flatness test proved to be an efficient method of checking low dose profiles, much faster than any of the alternative methods. The MLC test was found to be extremely sensitive to sudden changes in MLC calibration but works best with a composite reference image consisting of an average of five successive days' images. When used in this way it proved an effective and efficient daily check of MLC calibration. Overall, the amorphous silicon EPID was found to be a suitable device for IMRT QC although it is not recommended for dosimetric tests. Automatic procedures for low monitor unit profile analysis and MLC leaf positioning yield considerable time-savings over traditional film techniques. PMID- 16370417 TI - Final Aperture Superposition Technique applied to fast calculation of electron output factors and depth dose curves. AB - The Final Aperture Superposition Technique (FAST) is described and applied to accurate, near instantaneous calculation of the relative output factor (ROF) and central axis percentage depth dose curve (PDD) for clinical electron beams used in radiotherapy. FAST is based on precalculation of dose at select points for the two extreme situations of a fully open final aperture and a final aperture with no opening (fully shielded). This technique is different than conventional superposition of dose deposition kernels: The precalculated dose is differential in position of the electron or photon at the downstream surface of the insert. The calculation for a particular aperture (x-ray jaws or MLC, insert in electron applicator) is done with superposition of the precalculated dose data, using the open field data over the open part of the aperture and the fully shielded data over the remainder. The calculation takes explicit account of all interactions in the shielded region of the aperture except the collimator effect: Particles that pass from the open part into the shielded part, or visa versa. For the clinical demonstration, FAST was compared to full Monte Carlo simulation of 10 x 10, 2.5 x 2.5, and 2 x 8 cm2 inserts. Dose was calculated to 0.5% precision in 0.4 x 0.4 x 0.2 cm3 voxels, spaced at 0.2 cm depth intervals along the central axis, using detailed Monte Carlo simulation of the treatment head of a commercial linear accelerator for six different electron beams with energies of 6-21 MeV. Each simulation took several hours on a personal computer with a 1.7 Mhz processor. The calculation for the individual inserts, done with superposition, was completed in under a second on the same PC. Since simulations for the pre calculation are only performed once, higher precision and resolution can be obtained without increasing the calculation time for individual inserts. Fully shielded contributions were largest for small fields and high beam energy, at the surface, reaching a maximum of 5.6% at 21 MeV. Contributions from the collimator effect were largest for the large field size, high beam energy, and shallow depths, reaching a maximum of 4.7% at 21 MeV. Both shielding contributions and the collimator effect need to be taken into account to achieve an accuracy of 2%. FAST takes explicit account of the shielding contributions. With the collimator effect set to that of the largest field in the FAST calculation, the difference in dose on the central axis (product of ROF and PDD) between FAST and full simulation was generally under 2%. The maximum difference of 2.5% exceeded the statistical precision of the calculation by four standard deviations. This occurred at 18 MeV for the 2.5 x 2.5 cm2 field. The differences are due to the method used to account for the collimator effect. PMID- 16370418 TI - Monte Carlo and thermoluminescence dosimetry of the new IsoSeed model I25.S17 125I interstitial brachytherapy seed. AB - Monte Carlo simulation and experimental thermoluminescence dosimetry were utilized for the dosimetric characterization of the new IsoSeed model I25.S17 125I interstitial brachytherapy seed. The new seed design is similar to that of the selectSeed and 6711 seeds, with the exception of its molybdenum marker. Full dosimetric data are presented following the recommendations in the Update of the AAPM Task Group 43 report (TG-43U1). A difference of 3.3% was found between Monte Carlo dose rate constant results calculated by air kerma strengths from simulations using a point detector and a detector resembling the solid angle subtended to the seed by the Wide Angle Free Air Chamber (WAFAC) in the primary standard calibration geometry. Following the TG-43U1 recommendations, an average value of lambdaMC = (0.929 +/- 0.014) cGy h(-1) U(-1) was adopted for the new seed. This value was then averaged with the measured value of lambdaEXP = (0.951 +/- 0.044) cGy h(-1) U(-1) to yield the proposed dose rate constant for the new seed that is equal to lambda = (0.940 +/- 0.051) cGy h(-1) U(-1). The Monte Carlo calculated radial dose function and two-dimensional (2-D) anisotropy function results for the new seed were found in agreement with experimental results to within statistical uncertainty of repeated measurements. Monte Carlo simulations were also performed for 125I seeds of similar geometry and dimensions for the purpose of comparison. The new seed presents dosimetric characteristics that are very similar to that of the selectSeed. In comparison to the most extensively studied Amersham 6711 seed, the new one presents similar dosimetric characteristics with a slightly reduced dose rate constant (1.5%). PMID- 16370419 TI - Diffuse boundary extraction of breast masses on ultrasound by leak plugging. AB - We propose a semiautomated seeded boundary extraction algorithm that delineates diffuse region boundaries by finding and plugging their leaks. The algorithm not only extracts boundaries that are partially diffuse, but in the process finds and quantifies those parts of the boundary that are diffuse, computing local sharpness measurements for possible use in computer-aided diagnosis. The method treats a manually drawn seed region as a wellspring of pixel "fluid" that flows from the seed out towards the boundary. At indistinct or porous sections of the boundary, the growing region will leak into surrounding tissue. By changing the size of structuring elements used for growing, the algorithm changes leak properties. Since larger elements cannot leak as far from the seed, they produce compact, less detailed boundary approximations; conversely, growing from smaller elements results in less constrained boundaries with more local detail. This implementation of the leak plugging algorithm decrements the radius of structuring disks and then compares the regions grown from them as they increase in both area and boundary detail. Leaks are identified if the outflows between grown regions are large compared to the areas of the disks. The boundary is plugged by masking out leaked pixels, and the process continues until one-pixel radius resolution. When tested against manual delineation on scans of 40 benign masses and 40 malignant tumors, the plugged boundaries overlapped and correlated well in area with manual tracings, with mean overlap of 0.69 and area correlation R2 of 0.86, but the algorithm's results were more reproducible. PMID- 16370420 TI - An evaluation of the TSE MR sequence for time efficient data acquisition in polymer gel dosimetry of applications involving high doses and steep dose gradients. AB - The use of magnetic resonance imaging as a readout method for polymer gel dosimetry commonly involves long imaging sessions, particularly when high spatial resolution is required in all three dimensions, for the investigation of dose distributions with steep dose gradients and stringent dose delivery specifications. In this work, a volume selective turbo spin echo (TSE) pulse sequence is compared to the established Carr-Purcell-Meiboom-Gill (CPMG) multiecho acquisition with regard to providing accurate dosimetric results in significantly reduced imaging times. Polyethylene glycol diacrylate based (PABIG) gels were irradiated and subsequently scanned to obtain R2 relaxation rate measurements, using a CPMG multiecho sequence and a dual echo TSE utilizing an acceleration (turbo) factor of 64. R2 values, plotted against corresponding Monte Carlo dose calculations, provided calibration data of PABIG gels dose response over a wide dose range. A linear R2 versus dose relationship was demonstrated for both sequences with TSE results presenting reduced dose sensitivity. Although TSE data were found to deviate from linearity at lower doses compared to CPMG data, a relatively wide dynamic dose range of response extending up to approximately 100 Gy was observed for both sequences. The TSE and CPMG sequences were evaluated with a brachytherapy irradiation using a high dose rate 192Ir source and a gamma knife stereotactic radiosurgery irradiation with a single 4 mm collimator helmet shot. Dosimetric results obtained with the TSE and CPMG are shown to compare equally well with the expected dose distributions for these irradiations. The 60 fold scan time reduction achieved with TSE implies that this sequence could prove to be a useful tool for the introduction of polymer gel dosimetry in clinical radiation therapy applications involving high doses and steep dose gradients. PMID- 16370421 TI - Evaluation of precalibrated implantable MOSFET radiation dosimeters for megavoltage photon beams. AB - We have studied the response of factory calibrated implantable MOSFET detectors to absorbed doses from 100 to 400 cGy. The average measured dose is quite close to the true delivered dose, with the standard deviation falling between 1.4 and 3.6%. The measured dose tends to be slightly underestimated for smaller doses, while it tends to be slightly overestimated for larger doses. Thus, although the calibration of the detector is most accurate for doses close to the calibration dose of 200 cGy, it may be used over the range of commonly used doses in fractionated radiotherapy. PMID- 16370422 TI - Energy dependence of response of new high sensitivity radiochromic films for megavoltage and kilovoltage radiation energies. AB - The purpose of this paper is to evaluate the energy dependence of the response of two new high sensitivity models of radiochromic films EBT and XR-QA. We determined the dose response curves of these films for four different radiation sources, namely, 6 MV photon beams (6 MVX), Ir-192, I-125, and Pd-103. The first type (EBT) is designed for intensity modulated radiation therapy (IMRT) dosimetry, and the second type (XR-QA) is designed for kilovoltage dosimetry. All films were scanned using red (665 nm) and green (520 nm) light sources in a charge-coupled device-based densitometer. The dose response curves [net optical density (NOD) versus dose] were plotted and compared for different radiation energies and light sources. Contrary to the early GAFCHROMIC film types (such as models XR, HS, MD55-2, and HD810), the net optical densities of both EBT and XR QA were higher with a green (520 nm) than those with a red (665 nm) light source due to the different absorption spectrum of the new radiochromic emulsion. Both film types yield measurable optical densities for doses below 2 Gy. EBT film response is nearly independent of radiation energy, within the uncertainty of measurement. The NOD values of EBT film at 1 and 2 Gy are 0.13 and 0.25 for green, and 0.1 and 0.17 for red, respectively. In contrast, the XR-QA film sensitivity varies with radiation energy. The doses required to produce NOD of 0.5 are 6.9, 5.4, 0.7, and 0.9 Gy with green light and 19, 13, 1.7, and 1.5 Gy with red light, for 6 MVX, Ir-192, I -125, and Pd-103, respectively. EBT film was found to have minimal photon energy dependence of response for the energies tested and is suitable for dosimetry of radiation with a wide energy spectrum, including primary and scattered radiation. XR-QA film is promising for kilovoltage sources with a narrow energy spectra. The new high sensitivity radiochromic films are promising tools in radiation dosimetry. PMID- 16370423 TI - Immobilization effect of air-injected blanket (AIB) for abdomen fixation. AB - A new device for reducing the amplitude of breathing motion by pressing a patient's abdomen using an air-injected blanket (AIB) for external beam radiation treatments has been designed and tested. The blanket has two layers sealed in all four sides similar to an empty pillow made of urethane. The blanket is spread over the patient's abdomen with both ends of the blanket fixed to the sides of the treatment couch or a baseboard. The inner side, or patient side, of the blanket is thinner and expands more than the outer side. When inflated, the blanket balloons and effectively puts an even pressure on the patient's abdomen. Fluoroscopic observation was performed to verify the usefulness of AIB for patients with lung, breast cancer, or abdominal cancers. Internal organ movement due to breathing was monitored and measured with and without AIB. With the help of AIB, the average range of diaphragm motion was reduced from 2.6 to 0.7 cm in the anterior-to-posterior direction and from 2.7 to 1.3 cm in the superior-to inferior direction. The motion range in the right-to-left direction was negligible, for it was less than 0.5 cm. These initial testing demonstrated that AIB is useful for reducing patients' breathing motion in the thoracic and abdominal regions comfortably and consistently. PMID- 16370424 TI - AEC for scanning digital mammography based on variation of scan velocity. AB - A theoretical evaluation of nonuniform x-ray field distributions in mammography was conducted. An automatic exposure control (AEC) is proposed for a scanning full field digital mammography system. It uses information from the leading part of the detector to vary the scan velocity dynamically, thus creating a nonuniform x-ray field in the scan direction. Nonuniform radiation fields were also created by numerically optimizing the scan velocity profile to each breast's transmission distribution, with constraints on velocity and acceleration. The goal of the proposed AEC is to produce constant pixel signal-to-noise ratio throughout the image. The target pixel SNR for each image could be set based on the breast thickness, breast composition, and the beam quality as to achieve the same contrast-to-noise ratio between images for structures of interest. The results are quantified in terms of reduction in entrance surface air kerma (ESAK) and scan time relative to a uniform x-ray field. The theoretical evaluation was performed on a set of 266 mammograms. The performance of the different methods to create nonuniform fields decreased with increased detector width, from 18% to 11% in terms of ESAK reduction and from 30% to 25% in terms of scan time reduction for the proposed AEC and detector widths from 10 to 60 mm. Some correlation was found between compressed breast thickness and the projected breast area onto the image field. This translated into an increase of the ESAK and decrease of the scan time reduction with breast thickness. Ideally a nonuniform field in two dimensions could reduce the entrance dose by 39% on average, whereas a field nonuniform in only the scanning dimension ideally yields a 20% reduction. A benefit with the proposed AEC is that the risk of underexposing the densest region of the breast can be virtually eliminated. PMID- 16370425 TI - Sampling size in the verification of manufactured-supplied air kerma strengths. AB - Quality control mandate that the air kerma strengths (S(K)) of permanent seeds be verified, this is usually done by statistics inferred from 10% of the seeds. The goal of this paper is to proposed a new sampling method in which the number of seeds to be measured will be set beforehand according to an a priori statistical level of uncertainty. The results are based on the assumption that the S(K) has a normal distribution. To demonstrate this, the S(K) of each of the seeds measured was corrected to ensure that the average S(K) of its sample remained the same. In this process 2030 results were collected and analyzed using a normal plot. In our opinion, the number of seeds sampled should be determined beforehand according to an a priori level of statistical uncertainty. PMID- 16370426 TI - Development of high quantum efficiency, flat panel, thick detectors for megavoltage x-ray imaging: an experimental study of a single-pixel prototype. AB - Our overall goal is to develop a new generation of electronic portal imaging devices (EPIDs) with a quantum efficiency (QE) more than an order of magnitude higher and a spatial resolution equivalent to that of EPIDs currently used for portal imaging. A novel design of such a high QE flat-panel based EPID was introduced recently and its feasibility was investigated theoretically [see Pang and Rowlands, Med. Phys. 31, 3004 (2004)]. In this work, we constructed a prototype single-pixel detector based on the novel design. Some fundamental imaging properties including the QE, spatial resolution, and sensitivity of the prototype detector were measured with a 6 MV beam. It has been shown that the experimental results agree well with theoretical predictions and further development based on the novel design including the construction of a prototype area detector is warranted. PMID- 16370427 TI - Modeling skin collimation using the electron pencil beam redefinition algorithm. AB - Skin collimation is an important tool for electron beam therapy that is used to minimize the penumbra when treating near critical structures, at extended treatment distances, with bolus, or using arc therapy. It is usually made of lead or lead alloy material that conforms to and is placed on patient surface. Presently, commercially available treatment-planning systems lack the ability to model skin collimation and to accurately calculate dose in its presence. The purpose of the present work was to evaluate the use of the pencil beam redefinition algorithm (PBRA) in calculating dose in the presence of skin collimation. Skin collimation was incorporated into the PBRA by terminating the transport of electrons once they enter the skin collimator. Both fixed- and arced beam dose calculations for arced-beam geometries were evaluated by comparing them with measured dose distributions for 10- and 15-MeV beams. Fixed-beam dose distributions were measured in water at 88-cm source-to-surface distance with an air gap of 32 cm. The 6 x 20-cm2 field (dimensions projected to isocenter) had a 10-mm thick lead collimator placed on the surface of the water with its edge 5 cm inside the field's edge located at +10 cm. Arced-beam dose distributions were measured in a 13.5-cm radius polystyrene circular phantom. The beam was arced 90 degrees (-45 degrees to +45 degrees), and 10-mm thick lead collimation was placed at +/- 30 degrees. For the fixed beam at 10 MeV, the PBRA- calculated dose agreed with measured dose to within 2.0-mm distance to agreement (DTA) in the regions of high-dose gradient and 2.0% in regions of low dose gradient. At 15 MeV, the PBRA agreed to within a 2.0-mm DTA in the regions of high-dose gradient; however, the PBRA underestimated the dose by as much as 5.3% over small regions at depths less than 2 cm because it did not model electrons scattered from the edge of the skin collimation. For arced beams at 10 MeV, the agreement was 1-mm DTA in the high dose gradient regions, and 2% in the low-dose gradient regions. For arced beams at 15 MeV, the agreement was 1 mm in the high-dose gradient regions, and in the low-dose gradient region at depth less than 2 cm, as much as 5% dose difference was observed. This study demonstrated the ease with which skin collimation can be incorporated into the PBRA. The good agreement of PBRA calculated with measured dose shows that the PBRA is likely sufficiently accurate for clinical use in the presence of skin collimation for electron arc therapy. To further improve the accuracy of the PBRA in regions having significant electrons scattered from the edge of the skin collimation would require transporting the electrons through the lead skin collimation near its edges. PMID- 16370428 TI - Concomitant GRID boost for Gamma Knife radiosurgery. AB - We developed an integrated GRID boost technique for Gamma Knife radiosurgery. The technique generates an array of high dose spots within the target volume via a grid of 4-mm shots. These high dose areas were placed over a conventional Gamma Knife plan where a peripheral dose covers the full target volume. The beam weights of the 4-mm shots were optimized iteratively to maximize the integral dose inside the target volume. To investigate the target volume coverage and the dose to the adjacent normal brain tissue for the technique, we compared the GRID boosted treatment plans with conventional Gamma Knife treatment plans using physical and biological indices such as dose-volume histogram (DVH), DVH-derived indices, equivalent uniform dose (EUD), tumor control probabilities (TCP), and normal tissue complication probabilities (NTCP). We found significant increase in the target volume indices such as mean dose (5%-34%; average 14%), TCP (4%-45%; average 21%), and EUD (2%-22%; average 11%) for the GRID boost technique. No significant change in the peripheral dose coverage for the target volume was found per RTOG protocol. In addition, the EUD and the NTCP for the normal brain adjacent to the target (i.e., the near region) were decreased for the GRID boost technique. In conclusion, we demonstrated a new technique for Gamma Knife radiosurgery that can escalate the dose to the target while sparing the adjacent normal brain tissue. PMID- 16370429 TI - Evaluation of a diode array for QA measurements on a helical tomotherapy unit. AB - A helical tomotherapy system is used in our clinic to deliver intensity-modulated radiation therapy (IMRT) treatments. Since this machine is designed to deliver IMRT treatments, the traditional field flatness requirements are no longer applicable. This allows the unit to operate without a field flatness filter and consequently the 400 mm wide fan beam is highly inhomogeneous in intensity. The shape of this beam profile is mapped during machine commissioning and for quality assurance purposes the shape of the beam profile needs to be monitored. The use of a commercial diode array for quality assurance measurements is investigated. Central axis beam profiles were acquired at different depths using solid water built-up material. These profiles were compared with ion chamber scans taken in a water tank to test the accuracy of the diode array measurements. The sensitivity of the diode array to variations in the beam profile was checked. Over a seven week period, beam profiles were repeatedly measured. The observed variations are compared with those observed with an on-board beam profile monitor. The diode measurements were in agreement with the ion chamber scans. In the high dose, low gradient region the average ratio between the diode and ion chamber readings was 1.000 +/- 0.005 (+/- 1 standard deviation). In the penumbra region the agreement was poorer but all diodes passed the distance to agreement (DTA) requirement of 2 mm. The trend in the beam profile variations that was measured with the diode array device was in agreement with the on-board monitor. While the calculated amount of variation differs between the devices, both were sensitive to subtle variations in the beam profile. The diode array is a valuable tool to quickly and accurately monitor the beam profile on a helical tomotherapy unit. PMID- 16370430 TI - Target motion measurement without implanted markers and its validation by comparison with manually obtained data. AB - For an effective radiotherapy the exact tumor location must be determined. The localization has to take into account patient's setup position as well as internal organ motion. Among the different localization methods, the use of a computer tomography (CT) scanner in the therapy room has been proposed recently. Achieving a CT with the patient on the therapy couch, a patient's treatment position is captured. We present a method to locate tumor considering internal organ motion and displacements due to respiration. We tested the method with prostate and lung patients. The method found the most probable tumor position as well as, for high-mobility tumors located in the lung, its trajectory during the respiratory cycle. The results of this novel method were validated by comparison with manually determined target position. PMID- 16370431 TI - Effect of beamlet step-size on IMRT plan quality. AB - We have studied the degree to which beamlet step-size impacts the quality of intensity modulated radiation therapy (IMRT) treatment plans. Treatment planning for IMRT begins with the application of a grid that divides each beam's-eye-view of the target into a number of smaller beamlets (pencil beams) of radiation. The total dose is computed as a weighted sum of the dose delivered by the individual beamlets. The width of each beamlet is set to match the width of the corresponding leaf of the multileaf collimator (MLC). The length of each beamlet (beamlet step-size) is parallel to the direction of leaf travel. The beamlet step size represents the minimum stepping distance of the leaves of the MLC and is typically predetermined by the treatment planning system. This selection imposes an artificial constraint because the leaves of the MLC and the jaws can both move continuously. Removing the constraint can potentially improve the IMRT plan quality. In this study, the optimized results were achieved using an aperture based inverse planning technique called direct aperture optimization (DAO). We have tested the relationship between pencil beam step-size and plan quality using the American College of Radiology's IMRT test case. For this case, a series of IMRT treatment plans were produced using beamlet step-sizes of 1, 2, 5, and 10 mm. Continuous improvements were seen with each reduction in beamlet step size. The maximum dose to the planning target volume (PTV) was reduced from 134.7% to 121.5% and the mean dose to the organ at risk (OAR) was reduced from 38.5% to 28.2% as the beamlet step-size was reduced from 10 to 1 mm. The smaller pencil beam sizes also led to steeper dose gradients at the junction between the target and the critical structure with gradients of 6.0, 7.6, 8.7, and 9.1 dose%/mm achieved for beamlet step sizes of 10, 5, 2, and 1 mm, respectively. PMID- 16370432 TI - Tilted plane Feldkamp type reconstruction algorithm for spiral cone beam CT. AB - An approximate image reconstruction method for spiral cone beam computed tomography (CT), called tilted plane Feldkamp type reconstruction algorithm (TPFR), is presented in this paper, which extends Feldkamp cone beam reconstruction algorithm to deal with its inaccuracy and artifact problems caused by large cone angle. This is done by tilting the reconstructing planes to minimize the cone angle and optimally fit the spiral segment of the source. The tilted plane image reconstruction requires reforming the three-dimensional projection data set for the tilted plane and application of Feldkamp algorithm to the reformed data set. Analytical and computational results can show that the image reconstruction performance of the proposed TPFR algorithm is superior to that of the Feldkamp reconstruction algorithm in the image quality, volume coverage speed, maximum achievable pitch value, and slice sensitivity profiles. Moreover, it provides more accurate image reconstruction than the existing two dimensional reconstruction algorithms. PMID- 16370433 TI - On the automated definition of mobile target volumes from 4D-CT images for stereotactic body radiotherapy. AB - Stereotactic body radiotherapy (SBRT) can be used to treat small lesions in the chest. A vacuum-based immobilization system is used in our clinic for SBRT, and a motion envelope is used in treatment planning. The purpose of this study is to automatically derive motion envelopes using deformable image registration of 4D CT images, and to assess the effect of abdominal pressure on the motion envelopes. 4D-CT scans at ten phases were acquired prior to treatment for both free and restricted breathing using a vacuum-based immobilization system that includes an abdominal pressure pillow. To study the stability of the motion envelope over the course of treatment, a mid-treatment 4D-CT scan was obtained after delivery of the third fraction for two patients. The planning target volume excluding breathing motion (PTV(ex)) was defined on the image set at full exhalation phase and transformed into all other phases using displacement maps from deformable image registration. The motion envelope was obtained as the union of PTV(ex) masks of all phases. The ratios of the motion envelope to PTV(ex) volume ranged from 1.3 to 2.5. When pressure was applied, the ratios were reduced by as much as 29% compared to free breathing for some patients, but increased by up to 9% for others. The abdominal pressure pillow has more motion restriction effects on the anterior/inferior region of the lung. For one of the two patients for whom the 4D-CT scan was repeated at mid-treatment, the motion envelope was reproducible. However, for the other patient the tumor location and lung motion pattern significantly changed due to changes in the anatomy surrounding the tumor during the course of treatment, indicating that an image-guided approach to SBRT may increase the efficacy of this treatment. PMID- 16370434 TI - [The dentist on the DVI team--interdisciplinary philosophy of the Interpol DVI team]. AB - Recent experiences from mass disasters have shown that a structured and uniform methodology is the only good way to come to quick and correct results in disaster victim identification. It is therefore mandatory that everyone included in such operations is not only conscious of the procedures, but also willing to apply them. This can only be achieved by providing adequate information and training to all personnel involved, tactical and technical. It seems thus not unreasonable that in the future, DVI Interpol would only be willing to work with accredited personnel and DVI teams, in order to optimize the results of disaster victim identification operations. PMID- 16370435 TI - [Determination of dental age]. AB - A review of the most commonly used dental age estimating techniques is generated. The most important issue for the forensic odontologist involved in dental age estimation is to employ as many of these methods as possible by performing repetitive measurements and calculations of different age-related parameters. That is the only way in order to try and establish reliable dental age estimations. In particular, a special chapter is attributed to the complex problem of determining the age of majority. PMID- 16370436 TI - [Analysis of bite marks]. AB - Precise photographs, supplemented by impressions are used to research and compare bite marks. Different materials for impressions are available to the forensic odontologist. Subsequently, the acquired information of the bite mark will be compared with the dentition of a suspect through photographs, intra- and extra oral examination and impressions. The overlay, which can be created using a variety of methods, allows the comparison between the bite mark and the dentition of the suspect. Among the most commonly used are hand-tracing of study models, wax bites or photocopies, or the radio-opaque wax bite method. Thanks to modern technology and the use of computers, analyses of bite marks have gradually become more accurate. Current research carried out at the Katholieke Universiteit Leuven will demonstrate whether the computer program ForensicIQ is able to offer an even more precise comparison between a given bite mark and the dentition of a suspect. By no means should bite mark analysis alone be allowed to lead to a guilty verdict, but it will offer the opportunity to exclude a suspect from a crime when the data do not correspond. PMID- 16370437 TI - [Child abuse: bite marks versus other types of lesions]. AB - The number of reported child abuse cases is increasing every year. This may indicate that the threshold of reporting child abuse has dropped or that the incidence of this kind of physical violence has increased in our society. Physicians, dentists, emergency care personnel, and educators are, because of their professional relation with children, in a privileged position for detecting and reporting signs and symptoms of child abuse and should play a crucial role in the protection of potential victims of abuse. PMID- 16370438 TI - [Manual facial reconstruction in forensic medicine]. AB - The forensic facial reconstruction is a technique, often used on skeletal remains, when other conventional techniques of identification can't be used. The technique consists of rebuilding the soft tissue on the skull in order to obtain an image of the victim for the purpose of triggering identification. This is accomplished by anatomically rebuilding the muscles and soft tissue on the skull and/or use tissue depth markers on specific spots on the skull. PMID- 16370439 TI - [Computer-assisted facial reconstruction: recent developments and trends]. AB - Three-dimensional (3D) craniofacial reconstruction can be a useful tool in the identification of an unknown body. The progress in computer science and the improvement of medical imaging technologies during recent years has had a significant impact on this domain. New facial soft tissue depth data have been obtained. New guidelines for facial feature properties such as nose projection, eye protrusion or mouth width, have been suggested, but also older theories and "rules of thumbs" have been critically evaluated based on digital technology. New fast, flexible and objective 3D reconstruction computer-based programs are in full development. Employing the newer technologies and permanently evaluating the obtained results will hopefully lead to more accurate reconstructions. PMID- 16370440 TI - [Two-and three-dimensional reconstructions in creating the Photofit]. AB - Facial two- or three dimensional reconstruction is a method of last resort trying to trigger indirect identification through primary recognition offering a new lead to personal identification. It may be used when odontological or physical data comparison, fingerprints, DNA or other conventional identification techniques have failed. PMID- 16370441 TI - [Expertise in the context of criminal law]. AB - The procedures of expert investigations in criminal law are different from those in civil law. Being an expert in criminal law investigations assumes thorough knowledge of dentistry, forensic investigations and of rules and procedures to follow. Adequate training and continuous education are mandatory. PMID- 16370442 TI - [Positive identification of a body found in a wood]. AB - An identification of unknown remains is presented as an example of an interdisciplinary forensic investigation. PMID- 16370443 TI - [Contributions of dentistry to physical anthropology: example of the medieval cemetery of Coxyde]. AB - The role of the odontologist is crucial for the study of archaeological material. The excavations at the site of the ancient abbey of the Dunes of Koksijde led to the discovery of more than a thousand burial places. It is possible to estimate the age of the skeletons by studying molar dental wear. This method is however less appropriate for the industrialised populations of the 21st century, because of the switch to more diverse and processed (less abrasive) food. PMID- 16370444 TI - Language, the literature, and the patient. PMID- 16370445 TI - Adjuvant hormonal therapy in early breast cancer. AB - For many years, tamoxifen has been the gold standard adjuvant hormonal therapy with the greatest impact in early breast cancer for both pre- and postmenopausal women. Tamoxifen-based adjuvant endocrine therapy and chemotherapy have together contributed substantially to the reduction in breast cancer mortality that has occurred in recent years. Over the past few years, the role of aromatase inhibitors has grown in prominence and they are now on the threshold of supplanting tamoxifen as the new gold standard adjuvant therapy for postmenopausal women with estrogen-receptor-positive disease. With extended use of oral antihormones such as tamoxifen, the role of ovarian suppression on the other hand has become less clear in the adjuvant setting. This article reviews the most important data regarding the various adjuvant hormonal treatments in the management of early breast cancer and will also give a brief overview of the role of these agents in the neoadjuvant setting. PMID- 16370446 TI - Evaluation and management of women with BRCA1/2 mutations. AB - Genetic counseling and testing for susceptibility to breast and ovarian cancer is often an integral component of management for women with a personal and/or family history of these malignancies. In this article, we will briefly review the function and genetic epidemiology of the two major susceptibility genes, BRCA1 and BRCA2. We will then address approaches to risk assessment for women at high risk with respect to the probability that they harbor a deleterious mutation in one of these genes, and the likelihood that they will develop cancer if such a mutation is identified. The process of genetic counseling and testing is discussed, including a summary of the potential benefits, limitations, and risks of testing as well as a summary of test result interpretation. We conclude with a review and appraisal of the various options for breast and ovarian cancer risk reduction and screening options for women with a BRCA1 or BRCA2 mutation. PMID- 16370447 TI - A step-by-step guide to breast self-examination. PMID- 16370448 TI - Psychiatric assessment and symptom management in elderly cancer patients. AB - The number of older adults in the general population continues to grow. As their numbers rise, the elderly and the management of their medical problems must be of increasing concern for health-care professionals. Within this older population, cancer is a leading cause of morbidity and mortality. Although many studies have looked at the psychiatric implications of cancer in the general population, few studies tackle the issues that may face the older adult with cancer. This article focuses on the detection and treatment of depression, anxiety, fatigue, pain, delirium, and dementia in the elderly cancer patient. PMID- 16370449 TI - Management of a superior sulcus (pancoast) tumor. PMID- 16370450 TI - Model-based color halftoning using direct binary search. AB - In this paper, we develop a model-based color halftoning method using the direct binary search (DBS) algorithm. Our method strives to minimize the perceived error between the continuous tone original color image and the color halftone image. We exploit the differences in how the human viewers respond to luminance and chrominance information and use the total squared error in a luminance/chrominance based space as our metric. Starting with an initial halftone, we minimize this error metric using the DBS algorithm. Our method also incorporates a measurement based color printer dot interaction model to prevent the artifacts due to dot overlap and to improve color texture quality. We calibrate our halftoning algorithm to ensure accurate colorant distributions in resulting halftones. We present the color halftones which demonstrate the efficacy of our method. PMID- 16370451 TI - Quantization of accumulated diffused errors in error diffusion. AB - Due to its high image quality and moderate computational complexity, error diffusion is a popular halftoning algorithm for use with inkjet printers. However, error diffusion is an inherently serial algorithm that requires buffering a full row of accumulated diffused error (ADE) samples. For the best performance when the algorithm is implemented in hardware, the ADE data should be stored on the chip on which the error diffusion algorithm is implemented. However, this may result in an unacceptable hardware cost. In this paper, we examine the use of quantization of the ADE to reduce the amount of data that must be stored. We consider both uniform and nonuniform quantizers. For the nonuniform quantizers, we build on the concept of tone-dependency in error diffusion, by proposing several novel feature-dependent quantizers that yield improved image quality at a given bit rate, compared to memoryless quantizers. The optimal design of these quantizers is coupled with the design of the tone-dependent parameters associated with error diffusion. This is done via a combination of the classical Lloyd-Max algorithm and the training framework for tone-dependent error diffusion. Our results show that 4-bit uniform quantization of the ADE yields the same halftone quality as error diffusion without quantization of the ADE. At rates that vary from 2 to 3 bits per pixel, depending on the selectivity of the feature on which the quantizer depends, the feature-dependent quantizers achieve essentially the same quality as 4-bit uniform quantization. PMID- 16370452 TI - Hardcopy image barcodes via block-error diffusion. AB - Error diffusion halftoning is a popular method of producing frequency modulated (FM) halftones for printing and display. FM halftoning fixes the dot size (e.g., to one pixel in conventional error diffusion) and varies the dot frequency according to the intensity of the original grayscale image. We generalize error diffusion to produce FM halftones with user-controlled dot size and shape by using block quantization and block filtering. As a key application, we show how block-error diffusion may be applied to embed information in hardcopy using dot shape modulation. We enable the encoding and subsequent decoding of information embedded in the hardcopy version of continuous-tone base images. The encoding decoding process is modeled by robust data transmission through a noisy print scan channel that is explicitly modeled. We refer to the encoded printed version as an image barcode due to its high information capacity that differentiates it from common hardcopy watermarks. The encoding/halftoning strategy is based on a modified version of block-error diffusion. Encoder stability, image quality versus information capacity tradeoffs, and decoding issues with and without explicit knowledge of the base image are discussed. PMID- 16370453 TI - A robust structure-adaptive hybrid vector filter for color image restoration. AB - A robust structure-adaptive hybrid vector filter is proposed for digital color image restoration in this paper. At each pixel location, the image vector (i.e., pixel) is first classified into several different signal activity categories by applying a modified quadtree decomposition to luminance component (image) of the input color image. A weight-adaptive vector filtering operation with an optimal window is then activated to achieve the best tradeoff between noise suppression and detail preservation. Through extensive simulation experiments conducted using a wide range of test color images, the filter has demonstrated superior performance to that of a number of well known benchmark techniques, in terms of both standard objective measurements and perceived image quality, in suppressing several distinct types of noise commonly considered in color image restoration, including Gaussian noise, impulse noise, and mixed noise. PMID- 16370454 TI - Optimal erasure protection strategy for scalably compressed data with tree structured dependencies. AB - This paper is concerned with the transmission of scalably compressed data sources over lossy channels. Specifically, this paper is concerned with packet networks or, more generally, erasure channels. Previous work has generally assumed that the source elements form linear dependencies. The contribution of this paper is an unequal erasure protection algorithm which is able to take advantage of scalable data with more general dependency structures. In particular, the proposed scheme is adapted to data with tree-structured dependencies. The source elements are allocated to clusters of packets according to their dependency structure, subject to constraints on packet size and channel code-word length. Given a packet cluster arrangement, source elements are assigned optimal channel codes subject to a constraint on the total transmission length. Experimental results confirm the benefit associated with exploiting the actual dependency structure of the data. PMID- 16370455 TI - On multirate optimality of JPEG2000 code stream. AB - Arguably, the most important and defining feature of the JPEG2000 image compression standard is its R-D optimized code stream of multiple progressive layers. This code stream is an interleaving of many scalable code streams of different sample blocks. In this paper, we reexamine the R-D optimality of JPEG2000 scalable code streams under an expected multirate distortion measure (EMRD), which is defined to be the average distortion weighted by a probability distribution of operational rates in a given range, rather than for one or few fixed rates. We prove that the JPEG2000 code stream constructed by embedded block coding of optimal truncation is almost optimal in the EMRD sense for uniform rate distribution function, even if the individual scalable code streams have nonconvex operational R-D curves. We also develop algorithms to optimize the JPEG2000 code stream for exponential and Laplacian rate distribution functions while maintaining compatibility with the JPEG2000 standard. Both of our analytical and experimental results lend strong support to JPEG2000 as a near optimal scalable image codec in a fairly general setting. PMID- 16370456 TI - Feature-based wavelet shrinkage algorithm for image denoising. AB - A selective wavelet shrinkage algorithm for digital image denoising is presented. The performance of this method is an improvement upon other methods proposed in the literature and is algorithmically simple for large computational savings. The improved performance and computational speed of the proposed wavelet shrinkage algorithm is presented and experimentally compared with established methods. The denoising method incorporated in the proposed algorithm involves a two-threshold validation process for real-time selection of wavelet coefficients. The two threshold criteria selects wavelet coefficients based on their absolute value, spatial regularity, and regularity across multiresolution scales. The proposed algorithm takes image features into consideration in the selection process. Statistically, most images have regular features resulting in connected subband coefficients. Therefore, the resulting subbands of wavelet transformed images in large part do not contain isolated coefficients. In the proposed algorithm, coefficients are selected due to their magnitude, and only a subset of those selected coefficients which exhibit a spatially regular behavior remain for image reconstruction. Therefore, two thresholds are used in the coefficient selection process. The first threshold is used to distinguish coefficients of large magnitude and the second is used to distinguish coefficients of spatial regularity. The performance of the proposed wavelet denoising technique is an improvement upon several other established wavelet denoising techniques, as well as being computationally efficient to facilitate real-time image-processing applications. PMID- 16370457 TI - Is image steganography natural? AB - Steganography is the art of secret communication. Its purpose is to hide the presence of information, using, for example, images as covers. We experimentally investigate if stego-images, bearing a secret message, are statistically "natural." For this purpose, we use recent results on the statistics of natural images and investigate the effect of some popular steganography techniques. We found that these fundamental statistics of natural images are, in fact, generally altered by the hidden "nonnatural" information. Frequently, the change is consistently biased in a given direction. However, for the class of natural images considered, the change generally falls within the intrinsic variability of the statistics, and, thus, does not allow for reliable detection, unless knowledge of the data hiding process is taken into account. In the latter case, significant levels of detection are demonstrated. PMID- 16370458 TI - Identification of a discrete planar symmetric shape from a single noisy view. AB - In this paper, we propose a method for identifying a discrete planar symmetric shape from an arbitrary viewpoint. Our algorithm is based on a newly proposed notion of a view's skeleton. We show that this concept yields projective invariants which facilitate the identification procedure. It is, furthermore, shown that the proposed method may be extended to the case of noisy data to yield an optimal estimate of a shape in question. Substantiating examples are provided. PMID- 16370459 TI - A wrapper-based approach to image segmentation and classification. AB - The traditional processing flow of segmentation followed by classification in computer vision assumes that the segmentation is able to successfully extract the object of interest from the background image. It is extremely difficult to obtain a reliable segmentation without any prior knowledge about the object that is being extracted from the scene. This is further complicated by the lack of any clearly defined metrics for evaluating the quality of segmentation or for comparing segmentation algorithms. We propose a method of segmentation that addresses both of these issues, by using the object classification subsystem as an integral part of the segmentation. This will provide contextual information regarding the objects to be segmented, as well as allow us to use the probability of correct classification as a metric to determine the quality of the segmentation. We view traditional segmentation as a filter operating on the image that is independent of the classifier, much like the filter methods for feature selection. We propose a new paradigm for segmentation and classification that follows the wrapper methods of feature selection. Our method wraps the segmentation and classification together, and uses the classification accuracy as the metric to determine the best segmentation. By using shape as the classification feature, we are able to develop a segmentation algorithm that relaxes the requirement that the object of interest to be segmented must be homogeneous in some low-level image parameter, such as texture, color, or grayscale. This represents an improvement over other segmentation methods that have used classification information only to modify the segmenter parameters, since these algorithms still require an underlying homogeneity in some parameter space. Rather than considering our method as, yet, another segmentation algorithm, we propose that our wrapper method can be considered as an image segmentation framework, within which existing image segmentation algorithms may be executed. We show the performance of our proposed wrapper-based segmenter on real-world and complex images of automotive vehicle occupants for the purpose of recognizing infants on the passenger seat and disabling the vehicle airbag. This is an interesting application for testing the robustness of our approach, due to the complexity of the images, and, consequently, we believe the algorithm will be suitable for many other real-world applications. PMID- 16370460 TI - Bidirectional labeling and registration scheme for grayscale image segmentation. AB - In this paper, we introduce a new image segmentation scheme that is based on bidirectional labeling and registration and prove that its segmentation performance is equivalent to that of the conventional watershed segmentation algorithm. The proposed bidirectional labeling and registration scheme, which we refer to as bidirectional labeling and registration scheme (BIDS), involves only linear scans of image pixels. It uses one-dimensional operations rather than the queues that are used in traditional segmentation algorithms, which are two dimensional problems. BIDS also provides unique labels for individual homogeneous regions. In addition to achieving the same segmentation results, BIDS is four times less computationally complex than the conventional watershed by immersion technique. PMID- 16370461 TI - Reversible data embedding into images using wavelet techniques and sorting. AB - The proliferation of digital information in our society has enticed a lot of research into data-embedding techniques that add information to digital content, like images, audio, and video. In this paper, we investigate high-capacity lossless data-embedding methods that allow one to embed large amounts of data into digital images (or video) in such a way that the original image can be reconstructed from the watermarked image. We present two new techniques: one based on least significant bit prediction and Sweldens' lifting scheme and another that is an improvement of Tian's technique of difference expansion. The new techniques are then compared with various existing embedding methods by looking at capacity-distortion behavior and capacity control. PMID- 16370462 TI - The contourlet transform: an efficient directional multiresolution image representation. AB - The limitations of commonly used separable extensions of one-dimensional transforms, such as the Fourier and wavelet transforms, in capturing the geometry of image edges are well known. In this paper, we pursue a "true" two-dimensional transform that can capture the intrinsic geometrical structure that is key in visual information. The main challenge in exploring geometry in images comes from the discrete nature of the data. Thus, unlike other approaches, such as curvelets, that first develop a transform in the continuous domain and then discretize for sampled data, our approach starts with a discrete-domain construction and then studies its convergence to an expansion in the continuous domain. Specifically, we construct a discrete-domain multiresolution and multidirection expansion using nonseparable filter banks, in much the same way that wavelets were derived from filter banks. This construction results in a flexible multiresolution, local, and directional image expansion using contour segments, and, thus, it is named the contourlet transform. The discrete contourlet transform has a fast iterated filter bank algorithm that requires an order N operations for N-pixel images. Furthermore, we establish a precise link between the developed filter bank and the associated continuous-domain contourlet expansion via a directional multiresolution analysis framework. We show that with parabolic scaling and sufficient directional vanishing moments, contourlets achieve the optimal approximation rate for piecewise smooth functions with discontinuities along twice continuously differentiable curves. Finally, we show some numerical experiments demonstrating the potential of contourlets in several image processing applications. Index Terms-Contourlets, contours, filter banks, geometric image processing, multidirection, multiresolution, sparse representation, wavelets. PMID- 16370463 TI - Statistical behavior of joint least-square estimation in the phase diversity context. AB - The images recorded by optical telescopes are often degraded by aberrations that induce phase variations in the pupil plane. Several wavefront sensing techniques have been proposed to estimate aberrated phases. One of them is phase diversity, for which the joint least-square approach introduced by Gonsalves et al. is a reference method to estimate phase coefficients from the recorded images. In this paper, we rely on the asymptotic theory of Toeplitz matrices to show that Gonsalves' technique provides a consistent phase estimator as the size of the images grows. No comparable result is yielded by the classical joint maximum likelihood interpretation (e.g., as found in the work by Paxman et al.). Finally, our theoretical analysis is illustrated through simulated problems. PMID- 16370464 TI - An information fidelity criterion for image quality assessment using natural scene statistics. AB - Measurement of visual quality is of fundamental importance to numerous image and video processing applications. The goal of quality assessment (QA) research is to design algorithms that can automatically assess the quality of images or videos in a perceptually consistent manner. Traditionally, image QA algorithms interpret image quality as fidelity or similarity with a "reference" or "perfecft" image in some perceptual space. Such "full-referenc" QA methods attempt to achieve consistency in quality prediction by modeling salient physiological and psychovisual features of the human visual system (HVS), or by arbitrary signal fidelity criteria. In this paper, we approach the problem of image QA by proposing a novel information fidelity criterion that is based on natural scene statistics. QA systems are invariably involved with judging the visual quality of images and videos that are meant for "human consumption." Researchers have developed sophisticated models to capture the statistics of natural signals, that is, pictures and videos of the visual environment. Using these statistical models in an information-theoretic setting, we derive a novel QA algorithm that provides clear advantages over the traditional approaches. In particular, it is parameterless and outperforms current methods in our testing. We validate the performance of our algorithm with an extensive subjective study involving 779 images. We also show that, although our approach distinctly departs from traditional HVS-based methods, it is functionally similar to them under certain conditions, yet it outperforms them due to improved modeling. The code and the data from the subjective study are available at. PMID- 16370465 TI - Fingerprinting protocol for images based on additive homomorphic property. AB - Homomorphic property of public-key cryptosystems is applied for several cryptographic protocols, such as electronic cash, voting system, bidding protocols, etc. Several fingerprinting protocols also exploit the property to achieve an asymmetric system. However, their enciphering rate is extremely low and the implementation of watermarking technique is difficult. In this paper, we propose a new fingerprinting protocol applying additive homomorphic property of Okamoto-Uchiyama encryption scheme. Exploiting the property ingenuously, the enciphering rate of our fingerprinting scheme can be close to the corresponding cryptosystem. We study the problem of implementation of watermarking technique and propose a successful method to embed an encrypted information without knowing the plain value. The security can also be protected for both a buyer and a merchant in our scheme. PMID- 16370466 TI - Digital watermarking robust to geometric distortions. AB - In this paper, we present two watermarking approaches that are robust to geometric distortions. The first approach is based on image normalization, in which both watermark embedding and extraction are carried out with respect to an image normalized to meet a set of predefined moment criteria. We propose a new normalization procedure, which is invariant to affine transform attacks. The resulting watermarking scheme is suitable for public watermarking applications, where the original image is not available for watermark extraction. The second approach is based on a watermark resynchronization scheme aimed to alleviate the effects of random bending attacks. In this scheme, a deformable mesh is used to correct the distortion caused by the attack. The watermark is then extracted from the corrected image. In contrast to the first scheme, the latter is suitable for private watermarking applications, where the original image is necessary for watermark detection. In both schemes, we employ a direct-sequence code division multiple access approach to embed a multibit watermark in the discrete cosine transform domain of the image. Numerical experiments demonstrate that the proposed watermarking schemes are robust to a wide range of geometric attacks. PMID- 16370468 TI - Elastic body spline technique for feature point generation and face modeling. AB - Due to the advent of MPEG-4 standard, facial animation has been receiving significant attention lately. A common approach for facial animation is to use the mesh model. The physics-based transformation, elastic body spline (EBS), has been proposed to deform the facial mesh model and generate realistic expression by assuming the whole facial image has the same elastic property. In this paper, we partition facial images into different regions and propose an iterative algorithm to find the elastic property of each facial region. By doing so, we can obtain the EBS for mesh vertices in the facial mesh model such that facial animation can be more realistically achieved. PMID- 16370467 TI - Optimal block boundary pre/postfiltering for wavelet-based image and video compression. AB - This paper presents a pre/postfiltering framework to reduce the reconstruction errors near block boundaries in wavelet-based image and video compression. Two algorithms are developed to obtain the optimal filter, based on boundary filter bank and polyphase structure, respectively. A low-complexity structure is employed to approximate the optimal solution. Performances of the proposed method in the removal of JPEG 2000 tiling artifact and the jittering artifact of three dimensional wavelet video coding are reported. Comparisons with other methods demonstrate the advantages of our pre/postfiltering framework. PMID- 16370469 TI - Color demosaicking via directional linear minimum mean square-error estimation. AB - Digital cameras sample scenes using a color filter array of mosaic pattern (e.g., the Bayer pattern). The demosaicking of the color samples is critical to the image quality. This paper presents a new color demosaicking technique of optimal directional filtering of the green-red and green-blue difference signals. Under the assumption that the primary difference signals (PDS) between the green and red/blue channels are low pass, the missing green samples are adaptively estimated in both horizontal and vertical directions by the linear minimum mean square-error estimation (LMMSE) technique. These directional estimates are then optimally fused to further improve the green estimates. Finally, guided by the demosaicked full-resolution green channel, the other two color channels are reconstructed from the LMMSE filtered and fused PDS. The experimental results show that the presented color demosaicking technique outperforms the existing methods both in PSNR measure and visual perception. PMID- 16370470 TI - Motion-JPEG2000 codec compensated for interlaced scanning videos. AB - This paper presents an implementation scheme of Motion-JPEG2000 (MJP2) integrated with invertible deinterlacing. In previous work, we developed an invertible deinterlacing technique that suppresses the comb-tooth artifacts which are caused by field interleaving for interlaced scanning videos, and affect the quality of scalable frame-based codecs, such as MJP2. Our technique has two features, where sampling density is preserved and image quality is recovered by an inverse process. When no codec is placed between the deinterlacer and inverse process, the original video is perfectly reconstructed. Otherwise, it is almost completely recovered. We suggest an application scenario of this invertible deinterlacer for enhancing the sophisticated signal-to-noise ratio scalability in the frame-based MJP2 coding. The proposed system suppresses the comb-tooth artifacts at low bitrates, while enabling the quality recovery through its inverse process at high bitrates within the standard bitstream format. The main purpose of this paper is to present a system that yields high quality recovery for an MJP2 codec. We demonstrate that our invertible deinterlacer can be embedded into the discrete.wavelet transform employed in MJP2. As a result, the energy gain factor to control rate-distortion characteristics can be compensated for optimal compression. Simulation results show that the recovery of quality is improved by, for example, more than 2.0 dB in peak signal-to-noise ratio by applying our proposed gain compensation when decoding 8-bit grayscale Football sequence at 2.0 bpp. PMID- 16370471 TI - [Penicillin G and erysipelas--still being used?]. PMID- 16370472 TI - Childhood granulomatous periorificial dermatitis: a controversial disease. AB - The etiology, diagnosis and treatment of childhood granulomatous periorificial dermatitis (GPD) are highly controversial. Some authors underline the similarities between GPD and perioral dermatitis and consider both as part of a spectrum while other authors regard GPD as a distinctive condition. Clinically GPD is a papulo-pustular periorificial disease of the face which histopathologically shows a granulomatous perifollicular infiltrate. Because of its granulomatous pattern, GPD also has been related to cutaneous sarcoidosis. The clinical course is benign and self-limited. Topical steroids are regarded as either the main cause or a worsening factor. While topical treatment is occasionally effective, systemic antibiotic therapy of GPD is usually recommended. We report a "typical" case of GPD and review the literature to discuss the difficulties in its diagnosis and treatment. PMID- 16370473 TI - [Methicillin-resistant Staphylococcus aureus (MRSA) in chronic wounds: therapeutic options and perspectives]. AB - The contamination of chronic wounds with Methicillin-resistant Staphylococcus aureus (MRSA) represents a world-wide increasing problem. Although it is still unclear whether bacterial contamination is a relevant factor for delayed wound healing, verification of MRSA contamination has significant logistic consequences for the medical institution as well as for the patient. In particular, if MRSA contamination progresses towards a systemic infection, options for antibiotic therapy are greatly limited. In this article, we discuss current therapeutic standards and potential alternatives for eradication of MRSA. There is evident need for effective, novel approaches for elimination of MRSA from chronic wounds that avoid the development of bacterial resistance; otherwise therapeutic alternatives for antibacterial treatment of chronic wounds will become more limited. PMID- 16370474 TI - [Treatment of erysipelas in Germany and Austria--results of a survey in German and Austrian dermatological clinics]. AB - BACKGROUND: Erysipelas is a severe soft tissue infection usually caused by streptococci. The infection is restricted to the dermis and subcutaneous tissues. Treatment with antibiotics is essential. Many different therapeutic regimens are recommended, based mainly on empirical data and only partly proven by clinical studies. MATERIAL AND METHODS: Our aim was to evaluate the treatment of erysipelas in Germany and Austria by means of a questionnaire and to derive treatment recommendations from this data. RESULTS AND CONCLUSION: The majority of clinics treat patients with erysipelas as inpatients with intravenous antibiotics. The usual first line treatment is group G penicillin (80%). Other choices include amino-penicillins (11%), cephalosporins (16.5%) and anti staphylococcal penicillins (6.9%) are used. As second line antibiotics macrolides (63.5%), clindamycin (52.5%), penicillins (18.5%), cephalosporins (40%) and fluoroquinolones (20.5%) are mentioned. Carbapenems, tetracyclines, nitroimidazoles, glycopeptides, aminoglycosides, cotrimoxazole, fusidic acid and fosfomycin are used rarely. The median treatment duration is 10 days. Adjuvant measures are anticoagulation, non-steroidal anti-inflammatory agents, dressings, immobilization and treatment of local predisposing factors such as interdigital tinea. PMID- 16370475 TI - [Controlled layered removal of anogenital warts by argon-plasma coagulation]. AB - BACKGROUND: According to guidelines of the German STD Association, appropriate treatment of extensive anogenital warts with comparable recurrence rates includes cryotherapy, surgical excision, electrosurgery, CO2- and Nd:YAG-laser vaporisation. All these procedures are associated with varying degrees of risk for bleeding, release of potentially infectious aerosol, deep thermal destruction, slow wound healing, and scarring. METHODS: Using argon-plasma coagulation anogenital warts can be removed in layers in a controlled manner. High frequency current flows through the argon plasma to the tissue, allowing well-controlled, superficial tissue destruction. RESULTS: From January 2001 to March 2003, 54 patients with extensive genital, anal or anogenital warts were treated. After one treatment, 66% of the pa- tients remained disease-free in the following 4 months. Thirteen patients (24%) showed early recurrence after 4 weeks, five patients (9%) at a later date. In these patients, further treatment, in 9 cases combined with topical imiquimod cream, were necessary for complete remission. CONCLUSION: Compared to other therapeutic procedures, argon-plasma coagulation is a better controlled, quick and low-risk option for the treatment of anogenital warts. Depending on the type of involvement and individual risk factors, postoperative treatment with topical imiquimod cream may be useful. PMID- 16370476 TI - [Hyperpigmentosis and hypertrichosis of the eyelids after use of bimatoprost eye drops]. AB - After application of bimatoprost containing eye drops for four weeks, a 72 year old patient, suffering from glaucoma, developed hyperpigmentation and hypertrichosis of the eyelids. These symptoms can be seen in 1-10% of the patients as side effects in local therapy with bimatoprost, a prostaglandin analogue. The changes persist for varying periods of time but are reversible. Although prostaglandin analogues are sometimes more effective in the reduction of the intraocular pressure than beta-adrenoreceptor blocking agents, they remain second-line choices because of their side effects. This case report is meant to re-stimulate interest in the as yet obscure role of prostaglandins in hair biology, in clarifying the underlying mechanisms, and in exploring the possible therapeutic use of prostaglandin analogues in the management of hair loss (e.g. in androgenetic alopecia) or hair depigmentation (e.g. poliosis, canities). PMID- 16370477 TI - [Tacrolimus--a new therapeutic option for Hailey-Hailey-disease?]. AB - Hailey-Hailey disease is a chronic inflammatory skin disorder. Only symptomatic measures are available for long-term treatment. Topical tacrolimus has been shown to be effective in a number of inflammatory skin diseases, such as atopic eczema and can be used over long periods of time because of its safety profile. We tested tacrolimus ointment in a patient who had problems with Hailey-Hailey disease for many years. After only five days of treatment with topical tacrolimus, improvement was apparent. After four weeks of treatment, the pain and erosions were significantly ameliorated. The efficacy of topical tacrolimus in Hailey-Hailey disease may provide new insights into the pathogenesis of the disorder. PMID- 16370478 TI - [Desmoglein 1-negative, desmoglein 3-positive pemphigus herpetiformis with involvement of oral mucous membranes]. AB - A 46-year-old woman presented with a two year history of pruritic erythematous plaques with blisters, as well as oral erosions. Even though the cutaneous lesions fit best with dermatitis herpetiformis, bullous pemphigoid or pemphigoid gestationis, the histologic examination revealed eosinophilic spongiosis, most compatible with some form of pemphigus. The identification of intercellular IgG deposition on direct immunofluorescence and circulating IgG pemphigus antibodies on indirect immunofluorescence microscopy led to diagnosis of pemphigus herpetiformis. This rare form of pemphigus does not often involve the oral mucosa. In our patient, the explanation is that she had antibodies against desmoglein 3 but not desmoglein 1. Treatment with prednisolone and azathioprine caused rapid and complete healing. PMID- 16370480 TI - [Increased pigmentation of the sclera and periorbital region associated with increased cutaneous vascularization of the left arm]. PMID- 16370479 TI - [Therapeutic strategies for cutaneous sarcoidosis]. PMID- 16370481 TI - [Dermatologic in-patient rehabilitation of atopic dermatitis in adults]. PMID- 16370482 TI - International Board certification dermatopathology: a worldwide effort to rise standards in dermatopathology. PMID- 16370483 TI - Two unstable beta chain variants associated with beta-thalassemia: Hb Miami [beta116(G18)his-->Pro], and Hb Hershey [beta70(E14)Ala-->Gly], and a second unstable Hb variant at 170: Hb Abington [beta70(E14)Ala-->Pro]. AB - We report on three previously undescribed unstable hemoglobin (Hb) variants: Hb Miami, Hb Hershey and Hb Abington. Hb Miami was associated with a beta(+) thalassemia (thal) mutation [IVS-I-110 (G-->A)], whereas Hb Hershey was associated with a B0-thal mutation [IVS-I-1 (G-->A)]. Hb Hershey also has decreased oxygen affinity. These three Hb variants illustrate the range of clinical severity that can be seen with unstable Hb variants, particularly when combined with a thalassemic mutation. PMID- 16370484 TI - Hb Stara Zagora: a new hyper-unstable hemoglobin causing severe hemolytic anemia. AB - We describe a new hyper-unstable beta chain variant (codons 137-139, -6 bp) in a 2-year-old Bulgarian boy. The abnormal hemoglobin (Hb) is associated with severe hemolytic anemia as a consequence of its hyper instability. The child was admitted to the Pediatric Clinic (Faculty of Medicine, Stara Zagora, Bulgaria) at the age of 2 months. Because of anemia (Hb 6.9 g/dL) and high serum iron level (58 microM/L) the child was transfused. However, a month later his Hb level had dropped to 7.5 g/dL, and since then he has been on a regular monthly blood transfusion regimen. Hemoglobin analysis of a blood sample collected 2 months after the last transfusion at the age of 2 years, revealed no abnormalities except for the presence of inclusion bodies after incubation of peripheral blood with brilliant cresyl blue. Sequencing of the beta-globin gene revealed heterozygosity for a 6 bp deletion (-TGGCTA) at codons 137 [the second and third base pair (bp)], 138 and 139 (the first bp), forming a new codon at position 137 (GAT). This event eliminates three amino acids (Val-Ala-Asn) and introduces a new residue (Asp). It creates a new restriction site for HphI. The parents and his dizygotic twin brother had no history of hemolysis. The paternity of the child was confirmed by DNA analysis. PMID- 16370485 TI - Hb Amsterdam [alpha32(B13)Met--Ile (alpha2)]: a new unstable variant associated with an alpha-thalassemia phenotype and a new African polymorphism. AB - We have characterized a new abnormal hemoglobin (Hb) at position 32 of the alpha globin chain. The proband, a 38-year-old woman of Surinamese Black ancestry, was referred to the Academic Hospital in Amsterdam, The Netherlands, after 3 years of Prednisone treatment in Surinam. Kidney failure was diagnosed at the Nephrology Department, Free University Medical Center, Amsterdam, The Netherlands; the cortisone treatment was interrupted and dialysis was started. At this stage, a microcytic hypochromic anemia was observed with high reticulocyte (40%) and ferritin (500 microg/L) levels, and hemoglobinopathy was suspected. No abnormal bands were visible on alkaline electrophoresis and high performance liquid chromatography (HPLC). The Hb A2 level was normal (2.7%) and the erythrocyte count was low (3.59 x 10(12)/L) with a normal haptoglobin level (68 mg/100 mL). None of the common alpha-thalassemia (thal) deletion defects were present. The beta-globin gene sequence was normal but the alpha2-globin gene sequence revealed an ATG-->ATA transition at codon 32, changing the methionine into an isoleucine residue. The mutation, called Hb Amsterdam, was observed in the mother of the proband, who was also heterozygous for the--alpha3.7-thal deletion and affected by a moderate microcytic hypochromic anemia. Both Hb Amsterdam and the--alpha( 3.7) allele were found in association with a new polymorphism, IVS-I-39 (C-->T), previously observed in our laboratory in seven patients of African origin, on both the alpha1 and alpha2 genes. In addition, Hb Amsterdam was also associated with the common African alpha2 polymorphism (G-->CTCGGCCC at position 7238 and T- >G at position 7174). Hb Amsterdam is the first mutation ever described at codon alpha32, a position involved in alpha1/beta1 interaction. The possibility of a contribution of this mutation to the nephropatic state of the proband is discussed. PMID- 16370486 TI - First description in Tunisia of a point mutation at codon 119 (CCT-->TCT) in the alpha1-globin gene: Hb Groene Hart in association with the -alpha3.7 deletion. AB - Herein we describe the case of a Tunisian girl who presented with 3% Hb Bart's (gamma4) at birth. At the age of 3 years, she showed microcytosis and hypochromia in the absence of iron deficiency. The first step of molecular analysis was to test for the common Mediterranean mutations and the classical -alpha3.7 deletion was found in the heterozygous state. Since this finding could not explain the level of Hb Bart's at birth, or the hypochromia and microcytosis, all the alpha globin genes were sequenced. This revealed a rare point mutation at codon 119 (CCT-->TCT) in the alpha1-globin gene, identified for the first time in Tunisia, and which has previously been described as an unstable hemoglobin (Hb) variant named Hb Groene Hart [alpha119(H2)Pro-->Ser (alpha1)]. Here the alpha3.7/alpha(alpha)119(CCT-->TCT) genotype is responsible for the alpha thalassemia (thal) trait phenotype. PMID- 16370487 TI - Adult onset of a Thalassemia intermedia genotype in association with a -alpha-3.7 homozygosity. Hb G-Accra [beta73(e17)Asp-->Asn] in combination with beta- and alpha-thalassemia in the same family. AB - We present the case of a 39-year-old male of mixed Black and Chinese Surinamese origin referred because of abdominal pain and extreme tiredness. The patient reported that he had received a single blood transfusion in his youth and presented at intake with a severe microcytic hypochromic anemia. A chest X-ray and computer tomography (CT)-scan revealed bilateral mediastinal lymphadenopathy and interstitial infiltrates. Elevated Hb F (80%) and an unbalanced synthesis ratio (beta/alpha = 0.18) were compatible with severe beta-thalassemia (thal) intermedia. DNA analysis revealed a double heterozygoty for the -88 (C-->T) and the IVS-II- 654 (C-->T) mutations in the presence of a homozygosity for the alpha3.7 deletion. The two daughters of the proband were both heterozygous for the IVS-II-654 (C-->T) mutation and the -alpha3.7 deletion. The youngest daughter also carried the Hb G-Accra [beta73(E17)Asp-->Asn] mutation, inherited from the mother. Hb G-Accra, a mutant of presumed Ghanaian origin, described as non pathological in the carrier, is reported for the first time in combination with a severe fbeta(+)thal. The molecular background, haplotype of the mutations and a new A--> polymorphism at -309, 5' to the G(gamma) romoter, are reported. PMID- 16370488 TI - Alpha-thalassemia among tribal populations of Eastern India. AB - Five hundred and thirteen unrelated subjects belonging to various tribes of West Bengal, Arunachal Pradesh and Assam in Eastern India, were screened for the presence of alpha-thalassemia (thal) gene deletion(s) as a possible cause of unexplained anemia (Hb < 11 g/dL and/or MCH <28 pg, MCV < 78 fL). As reported earlier, beta-globin gene mutant alleles were found with a frequency of up to 20% in some tribes. In the present study, alpha-globin gene deletion alleles were found in 18% of subjects from West Bengal, 3.9% from Arunachal Pradesh and 3.84% from Assam tribesmen. Coexistence of alpha- and beta-globin gene abnormalities was observed in up to 18% of some tribal groups. The high inbreeding rate and lack of appropriate medical care make these populations particularly vulnerable. PMID- 16370489 TI - Hb taradale [beta82(EF6)Lys-->Arg]: a novel mutation at a 2,3-diphosphoglycerate binding site. AB - Hb Taradale [beta82(EF6)Lys-->Arg] was initially detected as a split Hb A0 peak on Hb A1c, monitoring. Red cell parameters, hemoglobin (Hb) electrophoresis and stability tests were normal. Mass spectrometry (ms) clearly identified a variant beta chain with a mass increase of 28 Da and peptide mapping located the mutation site to peptide betaT-9. DNA sequencing confirmed the presence of a novel beta82(EF6)Lys-->Arg mutation. This conservative substitution at a 2,3 diphosphoglycerate (2,3-DPG) binding site did not, however, appear to affect the P50 for oxygen binding. PMID- 16370490 TI - Identification of a new delta chain hemoglobin variant in a beta-thalassemia carrier: Hb A2-mumc [delta13(a10)Ala-->Asp]. AB - We describe a case of beta-thalassemia (thal) trait in which the patient also carries a novel delta chain variant due to a missense mutation at amino acid codon 13 (GCC-->GAC, Ala-->Asp). The level of Hb A2 was not elevated, raising the potential for misdiagnosis. PMID- 16370491 TI - The IVS-II-1 (G-->a) beta0-thalassemia mutation in cis with HbA2-Troodos [delta116(G18)Arg-->Cys (CGC-->TGC)] causes a complex prenatal diagnosis in an Iranian family. AB - The beta-thalassemia (thal) minor phenotypes with normal Hb A2 levels and decreased MCV and MCH values are relatively rare beta-thal traits. Here, we describe a family with normal Hb A2 and decreased MCV and MCH levels. Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) revealed the IVS-II-1 (G-->A) mutation in the beta-globin gene of the proband and her father. Direct sequencing of the gamma-globin gene of the proband and her father also revealed a previously reported variant called Hb A2-Troodos [gamma116(G18)Arg-->Cys] [in cis with the IVS-II-1 (G-->A) beta0-thal mutation]. This is the first case report of Hb A2-Troodos in association with the beta0 IVS II-1 mutation. Reduced Hb A2 expression by a concomitant Hb A2 beta-thal in cis or trans, may cause problems in carrier diagnostics, and eventually in genetic counseling and prenatal diagnosis when insufficient molecular analyses are performed. PMID- 16370492 TI - High Hb A2 beta-thalassemia due to a 468 bp deletion in a patient with Hb S/beta thalassemia. AB - We describe a case of Hb S/beta-thalassemia (thal) involving a 468 bp deletion that removes the beta-globin gene promoter but leaves the coding regions intact. This is the second report of this deletion, and our family study establishes that this deletion causes beta0-thal with unusually high levels of Hb A2 and Hb F. As with other genotypes involving deletions of the 5' region of the beta-globin gene, our patient had a mild form of Hb S/beta-thal. PMID- 16370493 TI - Characterization of a rare single alpha-globin gene deletion in a Chinese woman with Hb H disease. AB - A Chinese patient with Hb H (beta4) disease was found to be a compound heterozygote for a 2.4 kb alpha(+)-thalassemia (thal) deletion and the common Southeast Asian alpha0-thal deletion. The endpoints of the 2.4 kb deletion were identified by sequence analysis of the deletion junction. The deletion removes the entire alpha1-globin gene and leaves the alpha2-globin gene intact. PMID- 16370494 TI - A new G(gamma) chain variant: Hb F-Bron [gamma20(B2)val-->Ala]. AB - A new G(gamma) hemoglobin (Hb) variant, Hb F-Bron [gamma20(B2)Val-->Ala] on the first exon of the G(gamma)-globin gene is described. The variant was characterized by DNA sequencing and mass spectrometry (MS). Hematological abnormalities included hypochromia and microcytosis and were probably caused by an interaction with an alpha-thalassemia (thal) (3.7 kb) deletion in the heterozygous state. PMID- 16370495 TI - High incidence of Hb D-Los Angeles [beta121(GH4)Glu-->Gln] in Denizli Province, Aegean region of Turkey. AB - Denizli Province is located in the inner part of the Aegean region of Turkey and is one of the target areas for premarital screening. Here we report the abnormal hemoglobins (Hbs) observed during a premarital screening program in our region. According to our results, Hb D-Los Angeles [beta1211(GH4)Glu-->Gln (GAA-->CAA] (also known as D-Punjab, D-North Carolina, D-Portugal, Oak Ridge and D-Chicago), is the most frequent abnormal Hb in this region. PMID- 16370496 TI - G8 cash shortfall leaves AIDS fund in 'terrible trouble'. PMID- 16370497 TI - Rebuilding the trust. PMID- 16370498 TI - What are nurses worth? PMID- 16370499 TI - Classroom conflict. AB - While the emphasis in nurse training has shifted from the ward to the classroom, a rift has appeared between those who approve of a heavily academic approach and those who want a more hands-on system of learning. PMID- 16370500 TI - Welcome the friendly bacteria. AB - Recent research suggests that taking a probiotic and multivitamin and mineral supplement can reduce the misery of the common cold. But the findings remain controversial. The average adult has between three and four colds a year. Colds result in the loss of 150 million working days per annum in the UK. The cost of absenteeism to the economy is estimated to be pound 6.75 billion. About eight million people in the UK take multivitamins. PMID- 16370501 TI - Sexual history. AB - Sex advice is no new phenomenon, as a recent book by former Nursing Standard journalist John Naish reveals. PMID- 16370502 TI - A thorough overhaul. PMID- 16370503 TI - Reawakened trauma. PMID- 16370504 TI - Meeting the holistic needs of dependent older people. AB - AIM: To investigate whether national vocational qualifications (NVQs) have the potential to improve the quality of care for vulnerable older people. training METHOD: An antecedent study was carried out by interviewing care staff in care homes to determine the process of achieving an NVQ and to identify the specific areas of enquiry. Individual face-to-face interviews were conducted with representatives of six training providers, who were contracted by a county council to provide training and assessment to care staff undertaking NVQ in Care levels 2 and 3. Face-to-face interviews were also carried out with care staff working in residential and nursing homes, registered with the training providers. FINDINGS: The majority of care staff receive some training, usually in-house, but this training is not necessarily specific to NVQ. Achieving an NVQ in Care is not dependent on the ability to demonstrate competence in all aspects of care, because there is a choice of units for which candidates provide evidence. The extent to which the knowledge and skills of care staff are assessed, and the standards of care that they provide to meet the holistic needs of residents, depend largely on the competence of the assessor. NVQ in Care levels 2 and 3 are mainly concerned with personal care needs rather than the healthcare reguirements of residents. CONCLUSION: NVQ in Care is an assessment process and not a training course leading to a qualification. However, some training providers offer a set course, while others assess competence on knowledge and skills. The unit contents of NVQ in Care levels 2 and 3 do not address the holistic needs of older people because some important aspects of care, for example, enabling clients to eat, drink and use toilet facilities, are optional. PMID- 16370505 TI - Integrating risk management into working practice. AB - The aim of this article, the third in this series on the Seven Steps to Patient Safety (National Patient Safety Agency (NPSA) 2004), is to explore how risk management can be integrated into the culture of a healthcare organisation and the working practices of clinicians, managers and their immediate teams. PMID- 16370506 TI - Effective communication in palliative care. AB - This article focuses on the definitions of communication and an examination of their relationship to palliative care nursing. The underpinning theory is analysed as a means of understanding the communication process. The communication process in nursing is considered in the context of nurse/patient/family communication. While the focus of the article is on palliative care, the principles of communication as outlined also have relevance and applicability to nurses working in a variety of other clinical settings. PMID- 16370507 TI - Scar management. PMID- 16370508 TI - Networks pay off. PMID- 16370509 TI - Civic service. PMID- 16370510 TI - Second nature. PMID- 16370511 TI - Effects of 3-nitropropionic acid in rats: general toxicity and functional neurotoxicity. AB - 3-Nitropropionic acid (3-NP) causes biochemical and morphological alterations in human and animal brain. Young adult male Wistar rats received 3-NP i. p. on five consecutive days and were investigated four weeks later (subacute treatment). Acute effects were investigated 24 h after one i. p. dose. Spontaneous or stimulus-evoked activity was recorded from cortical sensory foci, from subcortical nuclei and from the tail nerve, in urethane anesthesia. The subacutely treated rats were dissected and organ weights measured to study general toxic effects. After subacute treatment, decrease was seen in the theta, and increase in the beta-2 and gamma, band of the spontaneous activity, dissimilarly in the cortical vs. subcortical sites. Latency of the sensory evoked potentials increased in all sensory foci after subacute treatment. Following acute treatment, amplitude of the somatosensory evoked potential decreased. The weight of the thymus decreased significantly in the treated rats. Further studies could elucidate the link between biochemical effects of 3-NP and the observed functional neurotoxic changes. PMID- 16370512 TI - Polychlorinated biphenyls in freshwater fish from the Zagreb area. AB - The aim of this study was to determine the levels of six polychlorinated biphenyls (referred to as PCB-28, PCB-52, PCB-101, PCB-138, PCB-153 and PCB-180, according to the IUPAC) in freshwater fish from the Zagreb area. A total of 216 samples was collected from five sites: from the Sava River upstream of Zagreb, in Zagreb, and downstream of Zagreb, from Lake Jarun, and from five fishponds from the Zagreb surroundings. Samples from all five sites contained polychlorinated biphenyls (PCBs). The maximal levels found in individual samples ranged between 20 microg kg(-1) and 50 microg kg(-1) depending on the congener. However, the total level of all examined congeners was far below the maximal allowed level of 2.0 mg kg(-1) ww, set by Croatian authorities for total PCBs. PMID- 16370513 TI - The effect of ochratoxin A on the concentration of protein carbonyls in rats. AB - The mechanism of ochratoxin A (OTA) toxicity has been found to involve the production of free radicals and consequent oxidative stress. Until now, the only studied pathway of OTA-caused oxidative damage was peroxidation of lipids. The aim of this study was to check whether OTA induced the production of protein carbonyls, markers of protein oxidation. Exposed rats (5 per group) were given daily OTA (0.5 mg kg(-1) b. w., i. p.) for 7 days, 14 days, or 21 days, and control rats solvent only (TRIS). All animals were killed 24 hours after the last treatment. The levels of OTA in plasma, kidney and liver homogenates increased gradually during the whole length of the experiment. The levels of protein carbonyls in kidney homogenates of OTA-treated rats was significantly higher after days 14 and 21 than in controls (P < 0.05). The levels of protein carbonyls in the liver was significantly higher in OTA-treated animals only after day 21 (P < 0.05). These results confirm that oxidative stress is involved in the mechanism of OTA toxicity, and that it causes the oxidation of proteins. PMID- 16370514 TI - Exercise-induced bronchoconstriction and exercise-induced respiratory symptoms in workers exposed to tea dust. AB - Assuming that airborne particles and pollutants are important contributing factors in the development of exercise-induced bronchoconstriction (EIB), we performed a case-control study including 63 tea workers (36 men and 27 women, aged 36-55, duration of employment 3-30 years) and an equal number of office workers, matched by sex and age. Exercise-induced respiratory symptoms were recorded in a questionnaire. Skin prick tests, spirometry, as well as exercise and histamine challenge were carried out. Environmental measurements were performed on site during the work shifts. The prevalence of self-reported exercise-induced respiratory symptoms and EIB did not differ significantly between the exposed and control group (41.6% vs 36.8%, and 6.4% vs 4.8%, respectively). In both exposed and control workers, EIB was strongly linked to asthma (P < 0.01). In the exposed workers it was significantly associated with positive family history of asthma (P < 0.01) and positive family history of atopies (P < 0.05), whereas in the exposed smokers it was significantly related to smoking duration (P < 0.05). Bronchial reaction to exercise in the exposed smokers was significantly greater than in control smokers (P < 0.05). Self reported exercise-induced respiratory symptoms were weakly associated with EIB, with a large proportion of false positive and a low proportion of false negative results in both groups. PMID- 16370515 TI - Analysis of dust samples from urban and rural occupational environments in Croatia. AB - This study estimated the exposure to dust mites in various occupational environments in Croatia. In total, 29 occupational dust samples were collected: 10 from urban areas (offices, archive of an insurance company, tobacco, paper recycling, fish-processing and textile plants, animal unit for experimental rats, winery), nine from rural areas (barley, hay, animal food and flour warehouses, tailor's shops, wood processing plant, swine confinement house, grocer's storeroom), and 10 samples from cabins of five fishing boats (five floor and five bed samples). Mites were microscopically identified, and the levels of Der p 1, Der f 1, and Der 2 allergens measured using the DUSTSCREEN test (Heska AG, Switzerland). Microscopy showed no mites in urban areas. Pyroglyphid mites (D. pteronyssinus) were found in all bed samples from fishing boats. Non-pyroglyphid mites were found in samples taken from barley, hay and animal food warehouses, the swine confinement house, grocer's storeroom, and fishing boats. Pyroglyphid mite allergens were detected in eight of 10 dust samples from the fishing boats. Median levels of Der p 1, Der f 1, and Der 2 in cabin bed samples were 10 microg g(-1), 0.2 microg g(-1), and 3.5 microg g(-1), respectively. Our findings on fishing boats suggests that pyroglyphid mites could be considered work-related allergens for fishermen. The results of this study confirmed non-pyroglyphid mites as occupational risk factors in various rural environments. PMID- 16370516 TI - Multidose activated charcoal in the treatment of carbamazepine overdose with seizures: a case report. AB - Serious complications after carbamazepine poisoning, such as coma, seizures, respiratory failure, cardiac conduction abnormalities, and death are more likely with serum levels greater than 170 micromol L(-1). We report a case of a single massive carbamazepine overdose in a 19-year-old male, following attempted suicide, without prior history of seizure disorder. On admission, three hours after ingestion, serum carbamazepine concentration was 179 micromol L(-1) and Glasgow Coma Scale scored 6. The patient was intubated and treated with multiple doses of activated charcoal for 48 hours. Twelve hours after ingestion, two repeated generalised myoclonic seizures were noted when serum carbamazepine levels peaked at 181 micromol L(-1), and were successfully treated with diazepam. Carbamazepine serum level fell within the therapeutic range 63 hours after ingestion and the patient was discharged without any long-term sequelae. As there is no antidote for carbamazepine poisoning, supportive treatment remains the only, but usually potent option. PMID- 16370517 TI - [Quick progress of a hearing impairment in a young plumber]. AB - Workers exposed to noise are regularly examined using tonal hearing tests. Of all noise-exposed worker population, about ten percent are hypersensitive to noise. Regular hearing tests conducted for two years revealed a hearing impairment in a young construction plumber who, according to on-site noise measurements, was occupationally exposed to permissible noise levels. Tonal hearing tests showed that the impairment was progressing quickly. This case study discusses probable causes of this lesion suffered by the subject, which is of the acoustic trauma type. PMID- 16370518 TI - [Beauvericin: chemical and biological aspects and occurrence]. AB - Beauvericin (BEA) is a cyclic hexadepsipeptide produced by Beauveria bassiana, Paecilomyces fumosoroseus, Paecilomyces tenuipes, Polyporus sulphurous, and a variety of Fusarium species. This mycotoxin shows antimicrobial, insecticidal, cytotoxic, and apoptotic activity. It is the most potent specific inhibitor of cholesterol acyltransferase and possesses ionophoric properties. BEA increases ion permeability in biological membranes by forming a complex with essential cations (Ca2+, Na+, K+), which may affect the ionic homeostasis. BEA has been frequently found in maize samples in Europe, USA and Africa and co-contamination with other Fusarium toxins such as fumonisins, and moniliformin was also found. There is only one report of BEA occurrence and co-occurrence with fumonisin B1, fumonisin B2 and ochratoxin A in Croatia. Biological activity of BEA may increase the toxicity of other mycotoxins that co-occur with BEA in food. The role of BEA in the development of human and animal mycotoxicosis is still unknown. PMID- 16370519 TI - [Preparations and biological properties of chiral compounds]. AB - Enantiomers of chiral compounds may express various biological activities and also different toxicities. Examples of different pharmacological effects of some chiral drugs such as fluoxetine, penicillamine, ibuprofen and albuterol are provided in this paper. Due to possible differences in activity, the chiral drugs are required to be pure enantiomeric compounds in order to be more effective and safer to use. In the laboratory, enantiomers are mainly synthesized as racemates (an equimolar mixture of enantiomers) while in biological pathways only one enantiomeric form is produced, such as amino acids, sugars and lipids. This paper presents the principles of chirality, general information about enantiomers and their biological aspects. It gives an outline of stereoselective methods for chromatographic resolution of enantiomers with stereoselective protein stationary phases, i.e. capillary electrochromatography (CEC) and high performance liquid chromatography (HPLC). The use of enzyme biotransformations (hydrolysis, oxidation and reduction) in chiral syntheses of carboxyl-, phosphoryl- or beta hydroxy esters, alcohols, epoxides and cis-carboxyl sulphoxide is described. This article also includes an example of lipase stereoselectivity improvement by amino acid mutations within the enzyme active site. PMID- 16370520 TI - Facts about nicotine toxicity. AB - Nicotine is an alkaloid obtained from the leaves of the tobacco plant, and it is the main constituent of tobacco smoke. This review opens with physical and chemical properties of nicotine and with general considerations about the methods for determining nicotine and its metabolite cotinine. It summarises the data about acute and long-term toxicity of nicotine and also reviews its metabolism and kinetic data, types of exposure and the main recognised health effects, with special attention to reproductive, cardiovascular, pulmonary, gastrointestinal, immunological and genetic toxicity. The main focus is on hazardous exposure and risk estimation. PMID- 16370521 TI - Alcohol and drink driving in the United Kingdom. AB - The background to and current status of the drink driving law in the United Kingdom is explained. Screening tests and analytical methods for alcohol in blood, breath and urine are described. Legal limits of 80 mg/100 mL in blood, 107 mg/100 mL in urine and 35 microg/100 mL in breath have been set. Penalties for exceeding the legal limit include a twelve month ban from driving for a first offence and three years and ten years respectively for subsequent convictions. Special reasons why a licence should not be withdrawn are considered together with the effect of medication on alcohol levels. Finally, the question of whether legal limits in the United Kingdom are too high is considered together with whether the punishments imposed are too severe. PMID- 16370522 TI - Research on surgical disorders in Ethiopia: challenges and prospects. PMID- 16370523 TI - Experience of surgical therapy in 72 patients with thoracic hydatidosis over a 10 year period. AB - OBJECTIVE: This review aims to evaluate retrospectively 72 patients diagnosed to have thoracic hydatid disease and treated surgically in Tikur Anbessa Hospital between December 1993 and November 2003. DESIGN: Retrospective Analysis hospital records. SETTINGS: Tikur Anbessa Hospital, Department of Surgery, Medical Faculty, Addis Ababa University, Addis Ababa, Ethiopia. PATIENTS AND METHODS: Medical records and Operation theater registers of seventy-two Patients operated on for Thoracic hydatidosis were evaluated retrospectively. Chest Roentgenogram was the main diagnostic tool used Main surgical techniques employed were evacuation in 44 (52.4%) cysts, cystotomy in 30 (35.7%) cysts, and enucleation in 10 (12%) cysts. Additional palliative procedures (such as pericystectomy and Capitonnage) and radical procedures (such as resection of the lung) were also employed. RESULTS: forty of the patients were male and thirty-two female with a mean age of 31.4 years. The most Common presenting symptoms were cough in 60 (83.3%) patients, expectoration in 47 (65.3%) patients and chest pain in 38 (52.8%) patients. Chest radiography revealed a round opacity or shadow in 58 (80.6%) patients. All patients were treated surgically. The mean hospital stay was 38.3 days. Major postoperative Complications occurred in 12 (16.9%) patients; pneumonia, pleural effusion and prolonged air leak were the complications in order of their frequency. There was one postoperative death and one recurrence at 6-months follow-up period. CONCLUSION: Surgical excision of pulmonary hydatidosis with maximum preservation of lung parenchyma is possible in the majority of patients and should remain the mainstay of surgical treatment. PMID- 16370524 TI - Perforated peptic ulcer in Tikur Anbessa Hospital: a review of 74 cases. AB - Little is known on the pattern of perforated peptic ulcer in Ethiopia. To evaluate the early, outcome of management, a five-year retrospective analysis of 74 operated cases of perforated peptic ulcer was undertaken. Perforated peptic ulcer accounted for 3.4% of the adult emergency surgical procedures. The mean age was 32.6 years, with a male to female ratio of 7.2 to 1.0. Fifty-six percent of the cases were unmarried. In nearly 22.0% of the patients, no previous history of peptic ulcer disease was documented. Delay in diagnosis was noted in 95% of the cases. Most patients had duodenal ulcer perforation, and about 78% had purulent peritonitis at laparotomy. Fourteen died in hospital. Early presentation of patients to surgical care facilities may reduce morbidity and mortality in cases of peptic ulcer perforation. PMID- 16370525 TI - Anti-HIV activity against immunodeficiency virus type 1 (HIV-I) and type II (HIV II) of compounds isolated from the stem bark of Combretum molle. AB - In vitro anti-HIV activity of various extracts prepared from the stem bark of Combretum molle (R. Br. Ex. G. Don.) Engl & Diels (Combretaceae), a plant widely used in Ethiopian traditional medicine for the treatment of liver diseases, malaria and tuberculosis has been assessed against human imnmuunodeficiencvy virus type 1 (HIV-1) and type 2 (HIV-2). The total extract was prepared by percolation with 80% methanol whilst the petroleum ether, chloroform, acetone and 100% methanol fractions were obtained by successive hot extraction using Soxhlet apparatus. Selective inhibition of viral growth was assessed by the simultaneous determination of the in vitro cytotoxicity of each of the extracts against MT-4 cells. Results obtained in this study indicate that the acetone fraction possessed the highest selective inhibition of HIV-1 replicatuon. Phytochemical investigation of the acetone fraction resulted in the isolation of two tannins and two oleanane-type pentacyclic triterpene glycosides. One of the tannins was identified as puncalagin (an ellagitannin), whilst the structure of the other (CM A) has not yet been fully elucidated. The saponins that were characterized as arjunglucoside (also called 4-epi-sericoside) and sericoside did not inhibit replication of either HIV-1 or HIV-2. On the other hand, both punicalgin and CM-A displayed selective inhibition of HIV-1 replication with selectrvitv indices (ratio of 50% cytotoxic concentration to 50% effective antiviral concentration) of 16 and 25, respectivelvy and afforded cell protection of viral induced cytopathic effect of 100% when compared with control samples. Neither of the tannins exhibited a selective inhibition to HIV-2 replication at nontoxic doses. PMID- 16370526 TI - Guillain-Barre syndrome in Ethiopian patients. AB - This is a ten years retrospective study conducted to assess the clinical profile and outcome of Guillain-Barre Syndrome (GBS) in Ethiopian patients. The medical records of all patients admitted with the diagnosis of GBS to the Department of Internal Medicine, Tikur Anbessa University Hospital, Addis Ababa, Ethiopia during the period September 1992 to September 2001 were reviewed. During the ten year review period ninety-five patients were admitted with the diagnosis of GBS, of which eighty-one met the National Institute of Neurological and Communicative Disorders and Stroke diagnostic criteria. The mean age (SD) of patients was 34.4 +/- 14.4 years. The male to female ratio was 1.25:1. History of antecedent event such as upper respiratory tract infection, diarrhea and vaccination was obtained in 47/81 (58.1%) of patients. The majority of the patients 62 (78.5%), presented with an ascending areflexic quadriparesis while 10 (12.7%) presented with a descending type of arefilexic quadriparesis, but in 6 (7.4%) of the patients the weakness was confined to the lower extremities (i.e. Paraparetic variant). Signs of autonomic involvement were seen in 25/81 (30.9%). Cytoalbzuminological dissociation in the cerebrospinal fluid was demonstrated in 62.3%. EMG was done in 47 patients The commonest electrophysiologic abnormality encountered was demyelinating picture in 26 (55.3%) followed by mixed and axonal in 12 (25.5%) and 9 (19.1%) respectively). Nineteen (70.3% of 27 GBS patients for whom serology for HIV was done were seropositive. The clinical findings were similar in both groups, except for a higher frequency of CSF pleocytosis, need for ventilatory support and mortality among HIV-positive patients. The overall mortality was 25.9%. This study highlights the importance of GBS as a cause of peripheral nerve disease in our setup. The higher mortality rate observed in this study appeared to be related mainly to the lack of adequate intensive carefacility and highlights the need to improve these services. Further-more, the finding of high frequency of seropositivity among GBS patients in this study underlines the need for further prospective research on the association and clinical course of GBS and HIV, particularly in Africa where HIV/AIDS has become an important public health problem. PMID- 16370527 TI - Outcome of corneal transplantation. AB - Corneal transplantation is a sight restoring surgery done for corneal blindness. The purpose of this retrospective preliminary study is to analyze the outcome of 54 corneal transplantations (for 32 females and 22 males) done in the Department of Ophthalmology between September 1998 and June 2002. The mean follow-up period was 13 months. Hundred percent (6/6) of the keratoconus (KC) and 85.4% of the nonkeratoconus grafts survived at a mean duration of 7.1 months and 16 months respectively. Seven of the 54 grafts (13%) have failed. The causes of graft failure were graft rejction in 4 and bacterial keratitis in 3 cases. Fifty of the 54 cases (92.6%) had a preoperative visual acuity of < 3/60. As a result of the transplantation, the percentage of blind eyes dropped from 92.6% to 21%. The data in this study confirms that corneal transplantation is a reasonably successful procedure in restoring sight for seleted cases of corneal blindness in Ethiopia. PMID- 16370528 TI - In vitro activity of Phytolacca dodecandra (Endod) against dermatophytes. AB - There is a need for better and less costly treatment of superficial mycotic infections, and one approach might be testing of'traditionally used plants for their antifungal activities as potentials for drug development. The aim of the present study was to determine the in vitro activity of Phytolacca dodecandra (Endod) on various species of dermatophytes and yeasts of medical importance. The aqueous extract from the plant P. dodecandra was tested for its antifungal activity against 33 dermatophyte and yeast strains. The minimum inhibitory concentration (MIC) was determined by adopting the broth microdilution method for testing of conidium forming filamentous fungi according to the NCCLS M38-P proposed standard with some modifications. The MIC of P. dodecandra against the dermatophytes tested ranged from 19.5 mg/l to 156.0 mg/l, while for all the yeasts the MIC was >500 mg/l. The minimum inhibitory concentration for 50% (MIC50) of the dermatophyte strains was 62.5 mg/l. The extract showed fungicidal activity against the dermatophytes that ranged from 19.5 to 312.5 mg/l. No activity was observed against the yeasts. From our preliminary results antifungal activity of P. dodecandra has been shown. This should he strengthened with more extensive studies which address both the antifungal activity and the active principle that is responsible for its fungicidal activity. PMID- 16370529 TI - How accurate is Siriraj stroke score among Ethiopians? A brief communication. AB - In hospitals where emergency neuroimaging is not available, the use of stroke scores has been proposed to distinguish ischemic from hemorrhagic stroke. The aim of this case study is to evaluate the accuracy of SSS among Ethiopian patients and was conducted in the Addis Ababa University teaching Hospital. Between September 1, 2000 and August 31, 2001. A total of 128 patients were admitted with a clinical diagnosis of stroke. 49(38.3%) had a CT scan result that was done within 24 to 72 hours of admission. CT revealed cerebral infarction in 20 (40.8%) patients and hemorrhagic stroke including subarachnoid hemorrhage in 29 (59.2%) patients. SSS was calculated and its sensitivity to diagnose hemorrhage and infarction was 77% and 61.5% respectively with an overall accuracy of 69.2% (Table 1) Its positive predictive values were 67% and 72.7% for hemorrhage and infarction respectively. The study has shown that bedside clinical examination does not always help in distinguishing between cerebral infarction and intracranial hemorrhage. PMID- 16370530 TI - Conjoined twins sharing major organs and with discordant congenital anomaly. AB - Thoraco-omphalopagus conjoined twins born to a 25-year-old para one Ethiopian lady are presented. Prenatal diagnosis was not made until elective cesarean section delivery was done for severe preeclampsia and twin pregnancy with breech presentation. Detailed autopsy results and relevant literature reviews are described. To the best of our knowledge, these are the second-documented cases of conjoined twins in this country. PMID- 16370531 TI - A case of symptomatic aberrant right subclavian artery. AB - Aberrant right subclavian artery is one of the aortic arch anomalies which is almost always asymptomatic. A 27-year-old lady, from Asebe Teferi, presented with dysphagia of eight months duration. Physical examination revealed no abnormality. Barium swallow and post Intravenous-contrast computerized tomography (CT) scan showed aberrant right subclavian artery. She was operated via the approach of left thoracotomy. The aberrant right subclavian was legated, brought anterior to trachea and reimplanted to the left stubclavian artery. She had uneventful postoperative course and discharged symptom free. PMID- 16370532 TI - Telepathology at Tikur Anbessa Hospital: how telemedicine works. PMID- 16370533 TI - Sleep in health and disease, an overview. Part II. PMID- 16370534 TI - Living by the code in clinical research. PMID- 16370535 TI - The distribution of mental disorders among an isolated island community in southern Ethiopia. AB - BACKGROUND: It is now recognized that surveys of unique communities are important to better understand the epidemiology of mental disorders. The Zeway islanders are completely different from the surrounding cultural groups, in the midst of whom they have been living in isolation for over three centuries. OBJECTIVES: To describe the prevalence and sociodemographic correlates of major mental disorders in this community. METHODS: Trained and supervised field workers were employed to conduct a door-to-door survey of the entire adult population of the islands using the Amharic CIDI (Composite International Diagnostic Interview). RESULTS: A total of 18.3% of the study subjects were classified as having ICD-10 diagnoses, excluding substance use disorders. The most prevalent CIDI/ICD-10 disorder in the study were neurotic and somatoform disorders (17%). Affective disorders were present in 2.2% while no cases of psychoses were found. The prevalence of tobacco use and alcohol dependence was 0.4% and 1.5%, respectively, with all cases being males. CONCLUSION: The pattern of occurrence of mental disorders in this isolated community is different from other populations in Ethiopia and elsewhere with no reported case of psychoses. PMID- 16370536 TI - Surgical management of carpal tunnel syndrome in neurosurgery practice in Ethiopia and review of literature. AB - Carpal tunnel compression of the median nerve is a common condition with an estimated incidence of 0.1% to 10% and a direct medical cost of above dollars 1 billion per year in the USA. Between the years 1995 to 2002, 28 cases, 6 (21.4%) males and 22 (87.6%) females of mild to severe forms of carpal tunnel syndrome (CTS) were seen and treated by the author in Tikur Anbessa Hospital (TAH) Neurosurgery Unit and a private hospital in Addis Ababa. Of the 28 cases, 22 (87.6%) patients, 7/22 (31.8%) mild and 15/22 (68.2%) severe forms, were surgically treated with the open surgery method. The male to female ratio was 1:3.7 and the age range 28-52 years, with a mean of 40.6 +/- 7.2 and the Confidence Interval of the mean 37.7-43.0; a median of 40 yrs and a mode of 40. Fifty percent of the cases were below 35 years and 75% below 46.75 years. The duration of illness was from 6 months to 8 years. There were 8/28 (28.6%) females with bilateral conditions. All the males had right sided carpal tunnel syndrome; the right hands were more involved in 23 (63.9%) cases while left hands only in 13 (36.1%) patients. All 22 patients were much relieved of the aching pains in the respective hands 3-5 days postoperatively and by 3 months 5 patients, the only ones who reported for follow-up, showed much reduced numbness and paraesthesiae with fairly good gripping capacity of the operated hands, though wasting and atrophy of the thenar eminences persisted requiring adequate exercise and early physiotherapy. There is no recurrence or worsening of the illness reported. Carpal tunnel syndrome is a treatable condition. It is stressed that an open surgery, required division of the flexor retinaculium and palmar aponeurosis extending to distal base attachment of the thenar eminence in the hand, is essential for the definitive treatment and to avoid any recurrence. Open surgery is easy and cheap and can be done by the general surgeon with a good anatomical knowledge of the hand. PMID- 16370537 TI - The pattern of orthopedic admissions in Tikur Anbessa University Hospital, Addis Ababa. AB - A one-year (April 1st 2003-March 31st 2004) prospective descriptive study of orthopedic admissions to Tikur Anbessa University Hospital (TAUH) was made in order to determine the burden of musculoskeletal conditions together with the present provision of care. A total of 824 patients were admitted of whom 576 were males and 248 females (M:F 2.3:1). The median age was 15 years (range 3 months to 80 years). Trauma was the cause for admission in 507 (61.5%), with road traffic accidents accounting for 211 (41.6%) and a fall for 195 (38.5%). 'Elective' patients waited for an average of 33.5 days for their admission. The mean duration of stay in hospital between admission and elective operation was 11.7 days. The post-operative stay was 20.6 days and the average length of hospital stay of all patients admitted was 35.4 days. The bed occupancy rate was 97.4% with an average time between the discharge of one patient and the admission of another (turnover interval) of 24 hours. Eighty-seven percent of the patients (97/112) who had previously visited a traditional healer presented with a chronic infection, a neglected or a complicated fracture or an advanced tumor. Eleven patients died (1.4%). Health education in the community and of traditional healers, together with a reduction of road traffic accidents would significantly reduce morbidity and mortality in Ethiopia. Major attempts must be made to reduce both the length of time patients wait for elective surgery after being admitted and their post-operative stay in order to serve more people. But it is vital to continue to teach conservative management of fractures as these will be the only methods available for the foreseeable future in hospital outside the centers of excellence. A series of specific recommendation are made to improve our care of orthopedic and trauma patients. We also suggest a broad based community study with an orthopedic input, which would help to produce a better profile and sound decisions to enable this to be achieved. PMID- 16370538 TI - Intestinal parasite isolates in AIDS patients with chronic diarrhea in Gondar Teaching Hospital, North west Ethiopia. AB - Chronic diarrhea is one of the major AIDS-defining illnesses in WHO Classification and occurs in 60-90% of HIV-infected patients in Africa. We did a case series study on parasite isolation in stool samples of AIDS patients with chronic diarrhea using wet-mount, Formol-Ether concentration technique and Modified Acid-Fast staining method in Gondar Teaching Hospital between January and September 2000. Seventy AIDS patients with chronic diarrhea were included in the study. Wasting syndrome was the clinical presentation in (97%) almost all AIDS patients who had chronic diarrhea. Intestinal parasites were detected in 41 out of 70 diarrheal specimens in AIDS patients. Multiple parasitic injections were detected in three diarrheal specimens. Intracellular parasite, (29%) Cryptosporidium parvum, and mucosal parasite, (17%) Strongyloides stercoralis were the frequently isolated parasites in diarrheal specimens of AIDS patients, accounting for 80% diarrhaegen pathogens among positive specimens. Cryptosporidium parvum, under-estimated cause of chronic diarrhea in immunocompetent adults, was found to be the prominent diarrheagen in AIDS patients in this study, similar with other studies in different African countries. PMID- 16370539 TI - Detection of intestinal microsporidiosis in diarrhoeal patients infected with the human immunideficiency virus (HIV-1) using PCR and Uvitex-2B stain. AB - A total of 105 single fresh stool samples were collected from diarrhoeal patients with (80 HIV-positive and 25 HIV-negative) from the Army and the Police hospitals, Addis Ababa. The stool samples were processed by water-ether sedimentation method; they were stained with Uvitex-2B technique for microscopic detection of intestinal microsporidium. A portion of all samples were preserved in 200microl PBS containing 2% PVPP ((Polyvinylpolypyrolidone) for confirmation with PCR. 18/105(17.2%) of the cases were positive for intestinal microsporidial infection by at least one method. 8/105 (7.6%) positive both by microscopy and PCR and 10/105 (9.5%) were positive only by PCR. All microsporidia positive cases were also HIV positive. Based on PCR analysis, 15 Enterocytozoon bieneusi and 3 Encephalitozoon intestinalis were identified. This study has shown that intestinal microsporidiosis is a common cause of chronic diarrhoea in advanced AIDS patients and this is mainly attributed to Enterocytozoon bieneusi. To the best of our knowledge, this is the first report of intestinal microsporidiosis in Ethiopia. It has an important implication for the understanding of the aetiology of diarrhoea in HIV/AIDS patients in the country. PMID- 16370540 TI - Prevalance and risk factors for yeast colonization in adult diabetic patients. AB - The aim of this study is to describe the associations between various host characteristics and yeast colonization; biofilm and phospholipase production in diabetic patients. The study was conducted between January 2003 and June 2003 in Abant Izzet Baysal University, Duzce, Turkey. One hundred and fourty five diabetic patients were included to the study. All oral and faecal specimens were placed on Sabourand dextrose agar with chloramphenicol and gentamicin. All isolates were identified with classic methods and carbohydrate assimilation patterns using API 20 CAUX. C. dubliniensis isolates were identified by CHROM agar Candida and chlamydospore formation according to the referral to the literature. Biofilm and phospholipase production was assessed by using previously described methods. The most common colonized species were C. albicans in oral and faecal cultures. C. dubliniensis was isolated in four oral cultures of the patients. Dental prosthesis, tooth brushing, older age, antibiotic use in the previous two weeks were found to be the significant factors for the oral yeast colonization. Younger age, smoking, shorter duration of diabetes, hospitalization in the last year and antibiotic use in the previous two weeks were found to be the significant factors for the faecal yeast colonization. Biofilm production was found to be positive in nine cases of oral and seven of faecal isolates. Phospholipase production was determined to be positive in 18 cases oral and 14 of faecal isolates. In conclusion, glycaemia control and other diabetic factors are not effective for yeast colonlizing. There was not any significant correlation between biofilm and phospholipase production and host characteristics in yeast colonization. Oral hygiene may be an effetive for decreasing the oral colonization in diabetic patients. PMID- 16370541 TI - In-vivo antipyretic studies of the aqueous and ethanol extracts of the leaves of Ajuga remota and Lippia adoensis. AB - BACKGROUND: There are quite large number of traditionally used plants that are used to treat ailments associated with fever in the ethnomedical system of Ethiopia. Most of them, however, have not been subjected to scientific investigation for their efficacy and safety. OBJECTIVES: To evaluate the antipyretic effects of the aqueous and ethanol extracts of the leaves of Ajuga remota and Lippia adoenesis. DESIGN: An experimental design in which five group of in-house bred albino mice weighing 30-35 g representing positive and negative control, and three dose levels extract treated groups, respectively was employed. The extracts, standard drugs and distilled water were administered into GIT by gavage to evaluate the antipyretic effect after inducing pyrexia in mice. RESULTS: All extracts of both plants that were administered orally at doses of 50, 100 and 200 mg per kg showed antipyretic property. The antipyretic effects were observed to be dose dependent. In both plants, the aqueous extract was found to have more potent antipyretic effect than the ethanol extract. No mouse manifested toxic effects at any dose levels of all the extracts for the observed period of two weeks. CONCLUSION: The present study show that both extracts of the investigated plants have got antipyretic effects and the extracts are effective and safe at the doses tested. This supports the traditional claim or use of these plants. PMID- 16370542 TI - Extracranial meningioma of the scalp: case report. AB - A 70-year-old man was admitted with a swelling on the left parietal region, which had grown progressively but slowly for three years. During the last six months the swelling increased in size relatively faster. The lesion was excised totally and sent for pathological examination. The histologic diagnosis was classical meningothelial meningioma. The specimen included some fragments of the skull, however, there was no evidence of an intracranial origin of the tumor. Eighteen months after surgery there was no evidence of recurrence or neurological deficit. Even though primary extracranial meningioma is a rare neoplasm, it should be included in the differential diagnosis of soft tissue tumors above all in the head and neck region. PMID- 16370543 TI - Should leptospira vaccination be encouraged? A case report. PMID- 16370544 TI - Sleep in health and disease. An overview. Part III. PMID- 16370545 TI - Assessment of quality of post abortion care in government hospitals in Addis Ababa, Ethiopia. AB - BACKGROUND: Unsafe abortion is the leading cause of maternal death in Ethiopia. It is also known to cause several acute and long-term complications leading to disabilities. In countries like Ethiopia where there is no access to safe abortion, improvement of the available postabortion care services is an important strategy. OBJECTIVES: The aim of the study is to assess the quality of health services with respect to postabortion care in hospitals in Addis Ababa, Ethiopia. METHODS: A cross sectional survey on quality of postabortion care was conducted among 422 postabortion patients attending four hospitals in Addis Ababa. Additionally, 42 service providers involved in providing postabortion care services were interviewed. Direct service delivery observation and inventory of equipment and supplies were also conducted. The data were analyzed using Statistical Package for Social Sciences. RESULTS: Interaction between service providers and patients was found to be satisfactory whereas information provision on important aspects of care such as danger signs and follow-up needs were very limited. Only about 20% and 3% received family planning counseling and contraceptive methods respectively. Other reproductive health related issues such as STIs and HIV/AIDS were rarely raised by the service providers during caring for the patients. Overall 92.3% of the patients responded that they were satisfied with the services they have received. CONCLUSION: The study has shown several missed opportunities and indicated important areas for future service delivery improvement. PMID- 16370546 TI - The distribution of Human Papilloma Virus infection in women with cervical histological abnormalities from an area with high incidence of cervical cancer. AB - BACKGROUND: Cancer of the cervix is the second most common cancer among women in the developing countries. Many epidemiological studies have shown that more than 90% of cancers of the cervix are attributed to infection by oncogenic human papilloma viruses. Though, the distribution of Human papilloma virus in a population has been studied worldwide, there is no single study done up to now on this subject in Ethiopia. OBJECTIVES: The aim of this study is to investigate the presence and distribution of Human Papilloma Virus infecting women with cervical abnormalities and furnish base line information to design future vaccination strategies suitable for the Ethiopian population. METHODOLOGY: A retrospective molecular pathologic analysis for Human Papilloma Virus was done on 284 formalin fixed and paraffin-embedded cervical biopsy specimens bearing different histopathologic abnormalities collected over five years period from the department of pathology of Gondar College of Medical Sciences, Gondar, Ethiopia. A polymerase chain reaction (PCR) based Human Papilloma Virus DNA detection and a nucleotide sequencing based Human Papilloma Virus typing was done at Max-Burger Research Institute, Leipzig, Germany. RESULTS: Human papilloma virus was found in 263 of the 284-biopsy specimens (92.60%) and Human Papilloma Virus type 16 was identified to be the most frequent genotype accounting for more than 76% of all HPV species. RECOMMENDATIONS: It is recommended that a wide population based epidemiological study is mandatory to define the exact picture of this disease in this country and a suitable vaccine targeting mainly HPV 16 will have a substantial impact on cervical cancer control in Ethiopia. PMID- 16370547 TI - Mental distress in medical students of Addis Ababa University. AB - Life in medical schools is said to be stressful leading to emotional distress. A study was conducted to estimate the prevalence of mental distress among medical students of Addis Ababa University using a Self Report Questionnaire (SRQ) in 2001. The SRQ was distributed to the entire student population through class representatives. A total of 273 (80%) students returned completed questionnaires. Of these, 83.2% were males and over 85% were above the age of 20 years. About 70% were Orthodox Christians and nearly half of the study population was from Addis Ababa. The one month prevalence of mental distress was found to be 32.6%. Over 6.0% reported that they had suicidal ideation in the last one month. Females reported symptoms of mental distress more often than males, but the difference was not statistically significant (X2 =2.97, df= 1, P=0.08). Age and level of training were shown to have significant association with mental distress (Age, X2=20.88, df=2, p<0.001 and year of study, X2 = 32.04, df=4, p<0.001). Those who were 20 years of age or below and those who were in the preclinical years of training were likely to report symptoms of mental distress more often than those who were older and above preclinical years of training, respectively. Students who use substances reported symptoms of mental distress more often than non users, but the difference was not statistically significant. The study showed that the risk of mental distress decreases as year of study advances in the medical school. However, this trend showed deflection in the year of internship. This study showed that mental distress is a common problem among medical students of Addis Ababa University. Further studies and support services for the students are recommended. PMID- 16370548 TI - Wound infection in Tikur Anbessa hospital, surgical department. AB - A prospective study of surgical wound infection has been conducted on 1754 surgical patients operated from January 1, 1999 to Dec 31, 1999 in Tikur Anbessa hospital. Demographic data and information on potential risk factors and the occurrence of wound infection in the first 30 days post operatively were collected using pretested data collection form. Among the patients 1162 (66.2%) were males and 592 (33.8%) were females. The age ranged from one day to 95 years with a mean of 30 (+/- 19) years. Seven hundred twenty eight (41.5%) wounds were classified as clean, 674 (38.4%) as clean-contaminated, 241 (13.7%) as contaminated and 111 (6.3%) as dirty and infected wounds. The overall wound infection rate was 14.8%. The infection rate for clean, clean-contaminated, contaminated and dirty and infected wounds were 8.0%, 14.8%, 22.0% and 44.2% respectively. The overall wound infection rates vary between 5.4% and 27.9% among consultant surgeons and it was 17.3% for residents. After adjusting for variation in patients condition and wound class, the infection rate of two surgeons was significantly higher, with odds ratio (OR) of 2.3 and 2.9. Shaving in the ward and use of drains were associated with a statistically significantly higher infection rate after adjusting for all factors considered in the study, with OR of 1.9 & 2.3 respectively. Preoperative antibiotics were found to be highly protective after multiple stepwise regressions with OR of 0.33. Post operative hospital stay and mortality were significantly higher in patients who had wound infection after adjusting for potentially confounding variables. It is concluded that the infection rate are much higher than the acceptable standard and this underscores the need for establishing hospital infection control system. Minimizing the use of drains, avoiding preoperative shaving, judicious use of preoperative antibiotics and meticulous surgical technique should be part of the immediate strategy to reduce wound infection rate. PMID- 16370549 TI - Meningitis in children beyond the neonatal period in Gondar University hospital. AB - Bacterial meningitis is one of the most serious infections in infants and children. The infection is associated with a high rate of complications. This is a retrospective study of 151 children with meningitis admitted to Gondar University hospital over a period of five years. There were 100 males and 51 females making male to female ratio 2:1. Forty seven (31.1%) of them were infants below the age of 12 months. Children under the age of 5 constituted for 88 (58.3%) of all the admissions. One hundred twenty five (82.8%) of them were discharged improved, 20 (13.2%) of them died in the hospital and the rest 6 (4.0%) left the hospital before completing their treatment. Most of the deaths 13/20 (65%) occurred in the first 72 hours of hospitalization. A significant number of patients 37 (25.5%) were treated with one or more antibiotics before admission. A higher case fatality rate was observed in those less than 6 months old, arrival after 48 hours after onset of illness, altered level of sensorium and in those having respiratory distress or seizure. Early diagnosis and prompt treatment is indicated to reduce mortality of acute bacterial meningitis. PMID- 16370550 TI - Development of resistance by Plasmodium falciparum to sulfadoxine/pyrimethamine in Amhara Region, Northwestern Ethiopia. AB - Following the high rate of chloroquine-resistant Plasmodium falciparum, Ethiopia changed the national drug policy in 1999 from chloroquine to sulfadoxine/pyrimethamine (SP) as first line. However, the useful therapeutic lifespan of SP may be limited by the rapid emergence of resistance. We conducted a study between October and November 2001 to examine the current extent of SP resistance to P. falciparum in Amhara Region, Northern Ethiopia. A total of 93 patients with uncomplicated P. faliciparum malaria were studied from Habru (n=50) and Weizazirt (n=43) localities. Drug resistance was evaluated using the 14-day WHO in vivo test protocol. Parasitological resistance to SP was found to be 32.0% (16/50) and 4.7% (2/43) in Harbu and Weizazirt localities, respectively. The corresponding clinical failure rates were 20.0% (10/50) and 4.7% (2/43). Of the parasitological failure at Harbu, 9 patients were classified as RI, 6 as RII and one as RIII type response. Among the clinical failures, 7 were LTF while the remaining 3 were ETF. Nevertheless, there was hardly any patient with RII/RIII or ETF response, and only two patients each with RI and LTF response were seen from Weizazirt locality. Therefore, the study underscores the presence of varying level of SP resistance to P. falicparum in the areas studied. Whereas SP remains quite effective in the treatment of uncomplicated P. falciparum in Weizazirt locality, the observed high rate of resistance from Harbu is alarming and an area of concern. PMID- 16370551 TI - Community diagnosis of common skin diseases in the Zay community of the Zeway Islands, Ethiopia. AB - BACKGROUND: Skin diseases are common public health problems throughout the world. Studies on skin problems are rare in Ethiopia. Easy and cheap methods of identifying skin conditions in the community are poorly developed in the country. OBJECTIVES: (1) To assess whether the newly developed Dermatological Screening Questionnaire (DSQ) can be used to effectively screen for common skin diseases and (2) to estimate the prevalence of skin diseases in a rural community. METHODS: A cross-sectional survey was conducted in the rural community residing on the islands of Lake Zeway. A locally prepared nine-item questionnaire (the DSQ) was used by lay-interviewers to screen for common skin diseases. Any respondent giving one or more positive responses, and a further 104 people who had replied negatively, underwent confirmatory diagnosis by specialist dermatologists. RESULTS: A total of 4697 people were included in the survey. Of these, 992 (20.0%) respondents screened positive on the DSQ. Out of 62.5% (N = 620) examined by dermatologists, 98.6% (n = 611) were found to have one or more skin conditions. Amongst screen negatives, 10.6% had skin diseases. Sensitivity and specificity of the DSQ was found to be 98% and 91%, respectively. The weighted prevalence of clinically-confirmed skin disease was 22.5%. Scabies was the most common of all skin conditions, followed by fungal infections. CONCLUSION: Our survey revealed that easily preventable and treatable skin diseases are common in this rural community. Nearly all those who gave a positive response to the DSQ had one or more skin disease confirmed on clinical examination. This finding indicates that the DSQ is a useful instrument for identification of common skin diseases in the community. PMID- 16370552 TI - Nasal NK/T-cell lymphoma causing diagnostic difficulties. AB - We present history, clinical presentation and anatomo-pathologic findings of a 24 year-old female patient with a nasal NK/T-cell lymphoma. This rare tumor is characterized by its angiocentric and angiodestructive growth, which results in extensive tumor necrosis. At the first encounter this tumor necrosis made it difficult to identify the nature of the tumor cells. However, this necrosis is a key feature: it is the result of the capacity of neoplastic NK/T-cells to invade vessels. The T-cell character of the neoplastic lymphoid has been shown by immunohistochemitry. PMID- 16370553 TI - A systematic review of nutritional rickets in Ethiopia: status and prospects. AB - BACKGROUND: Nutritional rickets contributes to the high burden of illness and death among Ethiopian children below 5 years of age. Lack of adequate information appears to have negatively influenced the impact of prevention and treatment programs. OBJECTIVES: The objective of this systematic review is to review existing body of knowledge about nutritional rickets in Ethiopia and develop recommendations that will guide development of strategies for effective interventions and research. METHODS: Published and unpublished information on nutritional rickets in Ethiopia was collected systematically. The information was critically reviewed and discussed in the context of regional and global situation. RESULTS: The major cause of nutritional rickets in Ethiopian children is lack of exposure to sunshine and/or inadequate intake of vitamin D. Lack of awareness and traditional beliefs are major causes for not exposing infants to sunshine. The disease is associated with poor socioeconomic status, low birth weight, protein-energy malnutrition and common childhood infections. Severe form of rickets is commonly seen at about 18 months of age. Rickets predisposes Ethiopian children to pneumonia. Diagnostic criteria used in most studies include two or more clinical signs and a characteristic radiological or biochemical abnormality. There are critical gaps in our knowledge about the epidemiology, ecology, and potentially effective interventions to prevent and treat rickets in Ethiopian children. CONCLUSION: Most of the studies on nutritional rickets in Ethiopia have been conducted in hospital settings. There is a need for well designed epidemiological and ecological studies. The biologic basis for the striking association between PEM and active rickets seen in Ethiopia needs to be determined The role of calcium deficiency, the part played by genetic factors, the nature of the association between the duration of breastfeeding and rickets, and the role of complimentary feeds in Ethiopian children need to be explored. Studies are required to determine the amount of sunshine required to prevent rickets in Ethiopian infants. Studies are required to establish criteria for the diagnosis of clinical and sub clinical rickets, particularly in malnourished children. Prevention programs need to be pursued consistently and systematically and treatment options, including the single massive dose of vitamin D, need to be re-evaluated and optimal mode of treatment determined. PMID- 16370554 TI - Circulating nitric oxide in women affected by weight loss amenorrhea during pulsatile gonadotropin-releasing hormone therapy. AB - No specific markers of the severity or prognosis of hypothalamic-pituitary gonadal axis disturbances associated with weight loss amenorrhea (WLA) are currently available. Circulating nitric oxide (NO), which is involved in the control of the reproductive function in women and is correlated with body mass index (BMI), at least in over-weight and obese subjects, might be a marker of the severity and/or progression of WLA. To test this hypothesis, we studied circulating NO levels in 11 women (age 27.1 +/- 1.59 yr) affected by WLA for 5.1 +/- 1.0 yr; in all patients hormonal therapy had been discontinued 10.0 +/- 3.15 months earlier. NO, determined by measuring its stable catabolite nitrite/nitrates (NOx), was compared with some clinical parameters and sex hormone levels. Subsequently, changes in NOx during pulsatile GnRH therapy (120 ng/kg bw sc every 120 min) were compared with the clinical and hormonal data. Fifteen normal women (27.3 +/- 1.6 yr) served as a control group. NOx was significantly lower (p<0.01) in WLA (8.8 +/- 2.0 micromol/l) than in control (18.7 +/- 2.5 micromol/l) subjects. No correlation between NOx and clinical parameters was noted in either WLA or control subjects. As a result of GnRH therapy, ovulatory cycles reappeared in 91% of WLA women. During the 1st cycle, periovulatory 17beta-estradiol levels were 110% higher than those noted in controls. During the 2nd cycle, NOx showed a slight increase in the follicular phase (+12% vs 1st cycle) followed by a drop during the luteal phase (-40% from the follicular phase); indeed, at that time, NOx correlated negatively with progesterone in both WLA (rS -0.32, p<0.05) and control (rS -0.48, p<0.05) subjects. NOx correlated with BMI at the time of the 2nd cycle (rS 0.71, p<0.05). In conclusion, this study shows that in WLA patients: 1) NO is low, as in other conditions of chronic anovulation; 2) it does not correlate with clinical data; 3) it takes longer than sex steroids to increase and show normal-like fluctuations; 4) its fluctuations are restored earlier in patients with greater BMI. PMID- 16370555 TI - C-reactive protein and tumor necrosis factor-alpha in gestational hyperglycemia. AB - OBJECTIVES AND STUDY DESIGN: Increasing evidences support an inflammatory origin for gestational hyperglycemia. This paper aims at investigating, cross sectionally and prospectively, the relationships between tumor necrosis factor alpha (TNF-alpha) and C-reactive protein (CRP) levels in normoglycemic and hyperglycemic pregnancies of women with and without conventional risk factors for gestational diabetes (GDM). RESULTS: Both at simple and multiple correlations TNF alpha levels are associated to fasting insulin, homeostasis model assessment insulin resistance (HOMA-IR) values and gestational hyperglycemia, while high sensitivity CRP (hsCRP) levels to body mass index (BMI). Furthermore, the TNF alpha levels of the second trimester and their increments in the third trimester are significant predictors of insulin levels measured at 32-36 weeks in the subgroup of hyperglycemic women with < or = 35 yr, BMI <25 kg/m2 and the absence of a first-degree relative with Type 2 diabetes (respectively, beta=1.1; 95%CI 0.66-1.48; p=0.002 and beta=1.0; 95%CI 0.36-1.66; p=0.02), in a multiple regression model, after multiple adjustments. In a second cohort of women at low risk for GDM (<25 yr, BMI <25 kg/m2 and absence of a first-degree relative with Type 2 diabetes), 24-28 weeks TNF-alpha levels are highly associated with corresponding insulin and HOMA values in the same model (respectively, beta=0.27; 95%CI 0.11-0.43; p=0.001 and beta=0.30; 95%CI 0.14-0.46; p<0.001). CONCLUSIONS: The data support the developing hypothesis that low-grade systemic inflammation is associated to GDM, in particular for pregnant women without conventional risk factors for gestational hyperglycemia, whose insulin resistance seems less explainable. PMID- 16370556 TI - Determinants of neurosurgical outcome in pituitary tumors. AB - OBJECTIVE: Neurosurgery is one of the main therapies for pituitary tumors; optimising outcome is highly desirable for the patient and the health system. We have analysed predictors of outcome in surgically treated pituitary adenomas operated in this centre. DESIGN AND PATIENTS: A total of 289 patients underwent neurosurgery for a pituitary tumor, by the same two neurosurgeons, between 1982 and 2001. Their records were examined to find predictors of post-surgical outcome. Thirty-eight percent were males, with a median age of 40.8 (8-82.7) yr; 51.9% had been operated since 1992, 92.2% by the transsphenoidal route. Most tumors (70.2%) were macroadenomas; 28.4% were non-functioning, 27.3% secreted PRL, 26.3% GH of which 14 (4.8%) also secreted PRL, 17.3% ACTH, 0.3% FSH and 0.3% TSH. RESULTS: A stepwise, forward logistic regression analysis revealed tumor size as the only significant predictor of radiological cure [odds ratio (OR) for macroadenoma 0.16 vs microadenoma, p=0.0005]. Hormonally, PRL-secretion by the tumor was a predictor of poor prognosis (OR 3.29 for cure of non-PRL-secreting tumors, p=0.005), as was tumor size (OR 0.45 for cure of macroadenomas, p=0.005). Considering simultaneous radiological and hormonal remission, tumor size (OR 0.35 for macroadenoma, p=0.0002), and operation date (OR 0.40 for up to 1991, p=0.0002) were the only significant predictors. CONCLUSIONS: PRL secretion, tumor size and operation date are the main predictors of neurosurgical outcome in pituitary tumors, the latter suggesting that neurosurgical experience plays an important role. PMID- 16370557 TI - DNA-flow cytometry of 207 pituitary adenomas: ploidy, proliferation, and prognosis. AB - The principal factors involved in pituitary adenoma formation are unknown. DNA flow cytometry is a useful study providing an estimation of a tumor proliferative rate. In this study, DNA-flow cytometry was performed to evaluate its capability to both assess prognosis and predict recurrence. Two hundred and seven fresh pituitary adenoma specimens were assessed by flow cytometry. Pre-operative endocrine function, previous medical treatment, radiographic appearance, surgical findings and immunohistochemistry were recorded for each patient. Patient outcomes were assessed at a mean follow-up of 5.3 +/- 3.1 yr. Endocrinologically inactive pituitary adenomas were predominantly euploids (50.8%). The highest proliferation rates occurred in Nelson's syndrome and the lowest in Cushing's disease. A significant difference in proliferation was observed with prolactinomas and acromegaly when a medical treatment was performed before primary surgery. Massive histological invasiveness was directly associated with a significant increase in proliferation rate. Radiotherapy did not affect the recurrence rate (4.4%) statistically. In conclusion, DNA-flow cytometry was found to be useful for determining ploidy and obtaining an overview of cell cycle status. It was helpful in identifying patients requiring closer follow-up, such as those with invasive adenomas and Nelson's syndrome. No single parameter revealed by DNA-flow cytometry could predict tumor prognosis or recurrence in the follow-up of 7.5 +/- 1.3 yr. PMID- 16370558 TI - Insulin therapy during Ramadan fast for Type 1 diabetes patients. AB - Patients with Type 1 diabetes (T1D) are normally exempt from the Ramadan fast; however, some patients insist on following the fast, often without the approval of their physicians. The aim of this study is to provide patients with T1D, who insist on fasting, with the most appropriate insulin regimen during the month of Ramadan. Seventeen patients with T1D who insisted on fasting were studied. Prior to Ramadan, the intermediate insulin was changed to ultralente in all patients. The total dose of insulin given to fasting patients by the end of Ramadan (45.7 +/- 14.4 U/day) was less than the total dose of insulin given before fasting (52.8 +/- 13.1 U/day) p<0.05. The ultralente and regular insulin constituted 70 and 30%, respectively, of the total insulin dose by the end of Ramadan, divided equally between Suhur (before sunrise) and Iftar (after sunset). There was no change in the glycosylated hemoglobin before and after fasting. Patients were instructed to break their fast after any episode of hypoglycemia. There were no severe daytime hypoglycemia episodes. We recommend that patients with T1D wishing to fast be switched to long acting insulin such as ultralente. The total insulin dose should consist of around 85% of their initial insulin dose and it should be composed of around 70% ultralente and 30% rapid insulin, divided equally between Suhur and Iftar. PMID- 16370559 TI - Ret proto-oncogene mutations in apparently sporadic Turkish medullary thyroid carcinoma patients: Turkmen study. AB - OBJECTIVE: Medullary thyroid carcinoma (MTC) frequently occurs in a sporadic form, but a substantial number of cases are hereditary and appear as part of the multiple endocrine neoplasia type 2 (MEN2) syndromes. Germline mutations in ret proto-oncogene have been shown to be the underlying cause of MEN2 syndromes. DESIGN: We carried out a multi-center study that aimed to perform mutational analysis of so called sporadic MTC patients. METHODS: Fifty-six MTC patients verified by histopathologic examination were subjected to genetic analysis. Exon 10, 11, 13, 14, 15 and 16 of the ret gene were analyzed by DNA sequencing and restriction enzyme digestion method. RESULTS: Among 56 apparently sporadic MTC patients, we identified 6 (10.7%) ret germline mutation carriers. Three individuals carried mutations at codon 634 in exon 11, one at codon 618 in exon 10, and two at codon 804 in exon 14. Identification of the predisposition gene mutation has allowed DNA-based strategy for direct mutation detection in patients with apparently sporadic MTCs. A substantial number of patients with apparently sporadic MTC carried germline mutations and 50% of their first degree relatives are expected to have or to develop MTC and/or other endocrine tumors. CONCLUSIONS: These results indicate the importance of careful genetic surveillance of any patient with apparently sporadic MTCs. PMID- 16370561 TI - Thyroid cancer in hyperthyroidism: incidence rates and value of ultrasound-guided fine-needle aspiration biopsy in this patient group. AB - Three hundred and thirty-three hyperthyroidism cases were retrospectively investigated to provide information about the association between hyperthyroidism and thyroid cancer. There were 112 cases of toxic multinodular goiter (TMNG), 77 cases of toxic nodular goiter (TNG) and 144 cases of Graves' disease (GD). All nodules detected in GD patients, all nodules greater than 1 cm diameter in nodular goiter patients, nodules 5-10 mm size diameter if they had calcification were fine-needle biopsied (FNAB) under ultrasound guidance (US-guided), and a total of 612 such biopsies were performed. The biopsy samples were cytologically assessed as benign (no.=552; 90.2%), suspicious (no.=6; 1.1%), malignant (no.=13; 2.1%), or inadequate for diagnosis (no.=41; 6.7%). All patients with a biopsy diagnosis of malignant or suspicious nodules underwent surgery. Histological examination confirmed the diagnosis of thyroid cancer in all 13 (2.1%) patients with malignant FNAB findings. Papillary thyroid carcinoma (PTC) was identified in 2 patients with TMNG (%1.8), 5 with TNG (%6.5) and 5 with GD (%3.5). Metastatic follicular thyroid carcinoma (FTC) was identified in a patient with TNG. Thyroid malignancy (micro- or macrocarcinoma) was diagnosed pre-operatively in all 13 cases by US-guided FNAB. Thyroid cancer was diagnosed in 6 (5.5%) of the 109 nodules detected in the TNG group, 2 (0.44%) of the 452 nodules detected in the TMNG group, and 5 (9.8%) of the 51 nodules detected in the GD group. Two (2.6%) of the 77 functioning nodules in the TNG patients were malignant, but none of the 402 functioning nodules in the TMNG patients was malignant. In patients with hyperthyroidism, US-guided FNAB is useful for detecting thyroid cancer in nodules greater than 5 mm diameter before radioiodine therapy or surgery. PMID- 16370560 TI - The vitamin D receptor fokI start codon polymorphism and bone mineral density in male hypogonadotrophic hypogonadism. AB - The genetic factors determining bone mineral density (BMD) are not well characterized. Many studies have investigated the relationship between the fokI polymorphism of vitamin D receptor (VDR) gene in diverse populations and gender, resulting in conflicting outcomes. Because peak bone mass in men is closely related to sufficient androgen release, the contribution of VDR gene on BMD might have been masked by hormonal status of adulthood. We therefore investigated the relationship between the fokI polymorphism of VDR and BMD in male patients with idiopathic hypogonadotrophic hypogonadism (IHH). Sixty-five untreated male patients with IHH and 39 healthy matched controls were evaluated. fokI polymorphism ("f" allele) was detected by polymerase chain reaction (PCR) restriction fragment length polymorphism using restriction endonuclease fokI, and BMD was measured by dual Energy X-ray absorpsiometry in lumbar spine, femur and radius. The distribution of FF, Ff, and ff alleles in patients with IHH and controls were not different (patients; 46%, 51%, 3% and controls; 51.3%, 46.1%, 2.6%, respectively). BMD levels in patients with IHH were significantly lower than controls. We categorized patients and control subjects in subgroups according to whether they had homozygous FF and heterozygous Ff genotype. No differences in BMD were seen between control subgroups, but total femur and femoral neck BMD were significantly lower in patients bearing heterozygous Ff genotype with IHH than homozygous FF ones (p=0.017 and p=0.009, respectively). Ff genotype might run down the BMD in cortical bone of femur, which needs to be proved in further studies. PMID- 16370562 TI - Undetectable inferior petrosal sinus levels of PTH-related peptide (PTHrP) in patients with ACTH-dependent Cushing's disease. AB - PTH-related peptide (PTHrP), a member of the PTH family, is widely expressed in foetal and adult tissues, and it has been found in benign and malignant tumors, including GH and PRL-secreting adenomas. Conflicting data are reported in literature on serum PTHrP concentrations in patients with Cushing's disease. The aim of the present study was to further evaluate peripheral and inferior petrosal sinus (IPS) serum PTHrP concentrations before and after CRH, in a group of consecutive patients with ACTH-dependent Cushing's disease. Nine patients with active ACTH-dependent Cushing's disease (8 women and 1 man, age +/- SD 41 +/- 13 yr) were submitted to peripheral and IPS sampling under fluoroscopic control before and after iv administration of CRH. All patients were subsequently submitted to transsphenoidal surgery and an ACTH-secreting microadenoma was found in all cases. In all patients, serum IPS and peripheral ACTH measurement were in keeping with the diagnosis of ACTH-dependent Cushing's disease. Serum PTHrP concentrations before and after CRH stimulation were below the sensitivity limit of the assay in all samples, and no gradient between IPS and peripheral sampling was observed. Our data, combined with others reported in literature, indicate that PTHrP release by ACTH-secreting tumors is not a common occurrence. Therefore, we conclude that IPS and peripheral PTHrP are of little clinical usefulness. PMID- 16370563 TI - Diagnosis of a case of Gitelman's syndrome based on renal clearance studies and gene analysis of a novel mutation of the thiazide-sensitive Na-Cl cotransporter. AB - Gitelman's syndrome is a recessively inherited renal tubular disorder characterized by low plasma potassium and magnesium levels, reduced calcium excretion, metabolic alkalosis, and increased plasma renin activity and plasma aldosterone concentration with normal blood pressure levels. A 23-yr-old man was referred to our department for further evaluation of hypokalemia. The patient also had hypomagnesemia and markedly reduced urinary calcium excretion. Renal clearance studies and gene analysis of the thiazide-sensitive Na-Cl cotransporter (TSC) were performed in the patient. In response to an iv injection of furosemide, chloride clearance (CCl) increased markedly, while distal fractional chloride reabsorption CH2O/(CH2O+CCl) was considerably reduced. In contrast, thiazide ingestion had no significant effects on these parameters. The patient had compound heterozygous mutations in the alleles encoding the TSC gene, one of which has not been formerly reported. Renal clearance studies and TSC gene analysis by amplification and direct sequencing are useful diagnostic tools for confirming a diagnosis of Gitelman's syndrome. PMID- 16370564 TI - Coexistence of Graves' disease and benign struma ovarii in a patient with marked ascites and elevated CA-125 levels. AB - BACKGROUND: Struma ovarii is the most common monodermal ovarian teratoma and consists mainly of thyroid tissue. Only 5% of patients with this tumor have features of hyperthyroidism. The pathophysiology of hyperthyroidism in struma ovarii is not clear. CASE: We describe a case of benign struma ovarii, presenting with the clinical features of an ovarian cancer: large complex pelvic mass, large amount of ascites and markedly elevated CA-125 serum levels. The patient was initially treated for Graves' disease, on the basis of ultrasonographic, laboratoristic and scintigraphic evidence. The resistance to the medical treatment led to thyroidectomy. After surgery the hyperthyroidism persisted and, suddenly, the patient presented ascites. A large pelvic mass was then diagnosed which, at the pathologic examination, was diagnosed as a struma ovarii. CONCLUSION: The struma ovarii always has to be considered when a pelvic mass is associated with features of hyperthyroidism. PMID- 16370565 TI - Isolated corticotrophin deficiency presenting with pericardial effusion. AB - Isolated ACTH deficiency is a rare disorder often presenting with long-standing aspecific symptoms combined with unusual clinical presentations. We here describe a patient in whom pericardial effusion was part of the dinical presentation and fully resolved on steroid at replacement dosage, highlighting the possibility of hypoadrenalism as an additional cause of pericardial effusion of unknown origin. PMID- 16370566 TI - Limited availability of nutritional vitamin D causing inappropriate treatment of vitamin D deficiency rickets with a response resembling pseudohypoparathyroidism type II in a Japanese patient. AB - Vitamin D deficiency rickets occasionally resembles pseudohypoparathyroidism type II (PHP type II) with respect to the response to exogenous PTH in the presence of hypocalcemia. We encountered a Japanese patient with stage 2 vitamin D deficiency rickets, who had increased urinary cAMP excretion and no response of urinary phosphate or N-acetyl-beta-D-glucosaminidase excretion to exogenous PTH under normocalcemic and normophosphatemic conditions, after treatment with 1,25(OH)2 vitamin D3. This case shows that it is possible for a response mimicking that of PHP type II to occur when the serum calcidiol level is low due to causes other than hypocalcemia and secondary hyperparathyroidism. When the serum calcidiol level is low, the appropriate treatment should be cholecalciferol or ergocalciferol. However, because neither is commercially available as a useful formulation in Japan, physicians are forced to inappropriately use calcitriol or analogs. PMID- 16370567 TI - Ghrelin and GHS on cardiovascular applications/functions. AB - Although initially recognised for their GH-releasing properties, the cardiovascular system has been recognised as a potentially important target for GH secretagogues (GHS). Moreover, a limited number of studies also indicate cardiovascular effects of ghrelin. So far reported cardiovascular effects of GHS and/or ghrelin include lowering of peripheral resistance, possible improvement of contractility and cardioprotective effects both in vivo and in vitro. Taken together, these results offer an interesting perspective on the future where further studies aiming at evaluating a role of GHS and ghrelin in the treatment of cardiovascular disease are warranted. PMID- 16370568 TI - Ghrelin in gastroenteric pathophysiology. AB - Ghrelin, an acylated peptide produced predominantly by the stomach, has been discovered to be a natural ligand of the growth hormone secretagogue receptor 1a (GHS-R1a). It is localized in distinct cells of the gastric mucosa, mainly distributed in the mid portion of the oxyntic gland characterized by P/D1 granules in man and X/A-like granules in rodents. The ghrelin cell represents the second most frequent endocrine cell type after the enterochromaffin-like cells in gastric oxyntic mucosa, pointing to a potentially relevant role in the physiology of the stomach. Ghrelin has no relevant homology with any known gastrointestinal peptide and displays strong GH-releasing activity both in animals and in humans. However, in addition to stimulating GH secretion, ghrelin possesses several other endocrine and extraendocrine biological activities that are explained by the widespread distribution of ghrelin and GHS-R1a expression. In the rat, ghrelin exerts a control in gastric acid secretion and motility: the gastric acid secretion is stimulated by peripheral administration of high doses of ghrelin, but inhibited by very low doses of ghrelin delivered into the central nervous system. Moreover, ghrelin provides a potent and dose-related gastroprotective action against ethanol- and stress-induced gastric ulcers. The integrity of both nitric oxide (NO) system and capsaicin afferent nerves are required for the gastroprotective effect of ghrelin, whereas the vagus nerve might be involved in conveying ghrelinergic signal from periphery to the brain. In addition, prostaglandins derived by the constitutive cyclooxygenase (COX) activity are essential for the protective activity of ghrelin in ethanol and stress-induced gastric lesions. Given its prevailing role in physiological and pathophysiological gastric function, the discovery of ghrelin will open new perspectives and potential clinical implications in the gastroenteric field. PMID- 16370569 TI - Ghrelin and the metabolic balance. AB - Several studies have provided evidence that ghrelin is involved in the regulation of metabolic balance. Recent data provide evidence that when studying the effects of ghrelin on energy balance, differential influences of the acylated and non acylated forms of the peptide must be considered. Although the correlation between the two forms is good, they may be different hormones with opposite actions with non-acylated ghrelin being able to antagonize some of the effects of the acylated form. Future studies will reveal which of the two forms of ghrelin, the acylated or non-acylated forms, dominates and determines the net influence on energy balance. PMID- 16370571 TI - Celiac disease in Type 1 diabetic children and adults: IgA class transglutaminase autoantibodies as the best screening marker. PMID- 16370572 TI - The medical riddle of the Great Sphinx of Giza. PMID- 16370570 TI - Type 2 diabetes mellitus in children and adolescents. AB - Over the last decade, there has been an alarming increase of Type 2 diabetes mellitus (T2DM) in youths, concomitant with the rise of obesity in this age group. T2DM is a progressive disease with a gradual increase in insulin resistance associated later with a decline in insulin secretion with fasting hyperglycemia. Prevalence of T2DM in children is mostly linked to some risk factors: obesity and sedentary lifestyle, puberty, membership of ethnic minorities, features of insulin resistance, family history of T2DM, female gender and perinatal factors. Prevention is essential and can be considered a public health approach directed to the general population. Treatment of T2DM in youth is complex and based on different strategies: diet, exercise and pharmacotherapy. An appropriated intervention program must be started early, in order to prevent or retard the progression of the disease and associated comorbidities. PMID- 16370573 TI - The husbandry and care of dendrobatid frogs. AB - Dendrobatid frogs are studied primarily for the bioactive alkaloids found in their skin. Also known as poison-dart frogs, these animals accumulate toxic alkaloids from dietary sources. The function and uses of the many alkaloids, the alkaloid accumulation system, and the basic biology and physiology of the frogs themselves are of research interest. Here we overview the taxonomy of these frogs and some of the unique aspects of their natural biology and reproduction. We also describe the components of a successful laboratory housing system, including temperature, lighting, humidity, ventilation, nutrition, health considerations, and handling. A brief summary of dendrobatid research highlights is provided. PMID- 16370574 TI - Effects of altered enclosure size and substrates on squirrel monkey (Saimiri sciureus sciureus) behavior. AB - We evaluated the effects of decreasing spatial density and changing substrate availability on activity, social spacing, and substrate use in a well established, social group of 12 squirrel monkeys (Saimiri sciureus sciureus). Using instantaneous scan samples, we recorded the specific activity, distance to nearest neighbor, and substrate type for each animal at 10-min intervals during daylight hours and at three variations in spatial density and substrate availability. Decreased spatial density was accompanied by significant increases in locomotion and mean interindividual distance. Contact behavior remained constant regardless of spatial density. There were no significant differences in time spent in activities with change in substrate availability, but mean interindividual distance decreased from 0.82 m to 0.69 m when more suspended log substrates were available but spatial density was constant. These results indicate that changes in preferred substrate availability may significantly affect social spacing in a captive primate social group. The importance of species substrate preferences should be considered to enhance group cohesiveness when captive enclosures are designed for nonhuman primates. PMID- 16370575 TI - Intraperitoneal administration of an iodine-based contrast agent to improve abdominal micro-computed tomography imaging in mice. AB - The purpose of this study was to estimate the optimal volume of an iodine-based contrast agent to administer to mice via intraperitoneal injection and the optimal time after injection to perform micro-computed tomography for maximal enhancement of abdominal organs. Eight mice were paired randomly; three pairs underwent imaging after receiving intraperitoneal injections of 125, 250, or 500 microl of contrast agent, and the fourth pair underwent imaging without receiving an injection. Each mouse was scanned three consecutive times, and each scan lasted 25 min so that we could observe the clearance of the contrast agent from the abdomen. We determined that introducing 250 microl of contrast agent into the abdominal cavity of the mice and then having the mice undergo micro-computed tomography 15 min after injection provided the optimal degree of contrast enhancement needed to distinguish the abdominal organs. These results may lead to expanded use of this imaging modality to assess abdominal organ margins in small animal studies in vivo. PMID- 16370576 TI - Transfusion via the carotid artery in the hamster. AB - Public health policy makers need quantitative scientific data to assess the risk to the blood supply posed by transmissible spongiform encephalopathy (TSE) diseases. To this end, our laboratory has developed a model of blood-borne TSE infectivity in hamsters infected with the 263K strain of scrapie, an experimental model of choice for quantitative studies of TSE infectivity. We report here a microsurgical method for cannulation of the carotid artery in the hamster that allows transfusion of a large fraction of the blood volume of the hamster, with virtually no blood loss to the surgical site or exposure to nervous tissue. Animals are minimally affected by the surgery, recover quickly and completely, and survive for their natural lives (as long as 3 years). This procedure has been used to obtain quantitative data on the transmissibility of the TSEs by transfusion, and these findings have informed public health policy on blood donation and blood use. PMID- 16370577 TI - Thyroparathyroidectomy procedures and thyroxine levels in the chinchilla. AB - Thyroid and embedded parathyroid glands were surgically removed (thyroparathyroidectomized) from adult chinchillas (Chinchilla laniger) to create an animal model of hypothyroidism. Thyroxine (T4) levels were measured at the time of surgery and one or two times after surgery from 10 thyroparathyroidectomized chinchillas and five sham controls to establish baseline serum T4 levels and to assess the degree and duration of hypothyroidism in this animal model. Baseline T4 levels ranged from 3.4 to 6.4 microg/dl (mean +/- 1 standard deviation, 5.25 +/- 0.84 microg/dl), with no differences between male and female chinchillas (5.4 +/- 0.6 microg/dl versus 5.2 +/- 1.0 microg/dl, respectively). T4 levels were significantly reduced in 80% of thyroparathyroidectomized chinchillas when measured 6 to 14 days after surgery, but reductions were variable, ranging from 9 to 89% in individual animals. There was rapid regrowth of thyroid tissue and a return of T4 levels to the baseline range in five of the seven animals followed for 1 to 2 months after surgery. T4 levels increased significantly in the sham-operated chinchillas, indicating a nonspecific effect of surgery. The results establish surgical procedures for creating a model of variable, transient hypothyroidism in the chinchilla. We also summarize published basal T4 values for various laboratory animals, to provide a convenient reference. PMID- 16370579 TI - Effects of barbiturate administration on hepatic and renal biochemical parameters in new zealand white rabbits. AB - To assess the initial response of various plasma hepatic and renal biochemical parameters to barbiturates, we assigned 30 new Zealand White rabbits to three treatment groups (n = 10 each): control (saline solution injected intravenously), pentobarbitone (30 mg/kg intravenously), and thiopentone (20 mg/kg intravenously). Blood samples were obtained from the central ear artery at six time points: before injection injection of the anesthetics or saline and at 10, 30, 60, and 120 min and 24 h afterward. Plasma alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma glutamiltransferase, blood urea nitrogen, and creatinine levels were measured using an autoanalyzer, and those of the treatment groups were compared with control group levels. The administration of thiopentone significantly increased plasma levels of alanine aminotransferase, aspartate aminotransferase, gamma glutamiltransferase and blood urea nitrogen, but that of plasma alkaline phosphatase significantly decreased. Plasma alkaline phosphatase and gamma glutamiltransferase levels significantly increased after pentobarbitone administration. From these results, we concluded that plasma levels of some hepatic and renal enzyme concentrations increase significantly within a short time after administration of thiopentone or pentobarbitone. Therefore, caution is required in interpreting data on plasma biochemical parameters from rabbits anesthetized with pentobarbitone or thiopentone. PMID- 16370578 TI - Lack of correlation of vaginal impedance measurements with hormone levels in the rat. AB - Hormone levels vary in female rats depending on estrous cycle stage. Vaginal cytology is a reliable method of staging female rats, but vaginal impedance offers an alternative depending on application. We sought to correlate vaginal impedance in cycling female rats with hormone levels. Vaginal cytology was the standard for comparison and verification of estrous cycle stage. Female rats (n = 41) were evaluated twice daily for 15 days via vaginal cytology and impedance to evaluate two or three estrous cycles per rat. During the last 5 days of the study, selected anesthetized sampling groups (n = 3 or 4 rats per group) were bled terminally at each time point to allow hormone determinations concurrently with vaginal cytology and impedance. Rats with abnormal vaginal smears or discharges (n = 5) were evaluated for reproductive tract histology. Rats classified in estrus by vaginal cytology had significantly higher vaginal impedance values than did nonestrus rats, but vaginal impedance and estrous cycle stage as determined by vaginal cytology did not correlate. Because of small sampling size in nonproestrus groups, correlation between vaginal impedance and hormone levels was evaluated only in proestrus rats (n = 22) and was nonsignificant. No correlation occurred between vaginal impedance and hormone levels in unstaged rats (n = 41). Two animals evaluated for reproductive tract histology showed evidence of pseudopregnancy. Vaginal impedance may be useful in distinguishing estrus from nonestrus rats but may be limited for chronic estrous cycle monitoring because of the possible risk of inducing pseudopregnancy. PMID- 16370580 TI - Limbic dermoid in a New Zealand White rabbit (Oryctolagus cuniculus). AB - Superficial keratectomy was performed in a New Zealand White rabbit for a suspected limbic dermoid. Histology confirmed the diagnosis. Ocular dermoids have been reported in a variety of laboratory animals. This is the first report of a corneal dermoid in rabbits. PMID- 16370581 TI - Spontaneous, generalized lipidosis in captive greater horseshoe bats (Rhinolophus ferrumequinum). AB - During a routine 6-month quarantine period, 3 of 34 greater horseshoe bats (Rhinolophus ferrumequinum) captured in mainland China and transported to the United States for use in echolocation studies were found dead with no prior history of illness. All animals were in good body condition at the time of death. At necropsy, a large amount of white fat was found within the subcutis, especially in the sacrolumbar region. The liver, kidneys, and heart were diffusely tan in color. Microscopic examination revealed that hepatocytes throughout the liver were filled with lipid, and in some areas, lipid granulomas were present. renal lesions included moderate amounts of lipid in the cortical tubular epithelium and large amounts of protein and lipid within Bowman's capsules in the glomeruli. In addition, one bat had large lipid vacuoles diffusely distributed throughout the myocardium. The exact pathologic mechanism inducing the hepatic, renal, and cardiac lipidosis is unknown. The horseshoe bats were captured during hibernation and immediately transported to the United States. It is possible that the large amount of fat stored coupled with changes in photoperiod, lack of exercise, and/or the stress of captivity might have contributed to altering the normal metabolic processes, leading to anorexia and consequently lipidosis in these animals. PMID- 16370582 TI - Acute paraplegia in a young adult long-evans rat resulting from T-cell lymphoma. AB - We describe an unusual case of acute paraplegia in a young adult (7.5-month-old) Long-Evans rat that resulted from a spontaneous T-cell lymphoma. At presentation, a neurologic exam revealed normal pelvic limb flexor reflexes, the absence of an anal reflex, and deep pain recognition. Radiographs did not identify any obvious spinal abnormality or osseous trauma, although the liver and spleen were prominent. Hematologic analysis disclosed leukocytosis with atypical lymphocytes. At necropsy, red, friable to gelatinous masses were found associated with the ventral aspect of the vertebral column at the levels of the thoracic and lumbar vertebrae. Impression smears of the mass revealed a monocytic cell population with cells averaging 7 to 10 microm in diameter and having scant cytoplasm and pleomorphic nuclei, characteristics consistent with a lymphoid neoplasm. Histologically, the neoplasm was unencapsulated, poorly demarcated and highly infiltrative, invading and effacing the bone marrow and epidural space of the vertebral column. Neoplastic cells also were identified in the femoral bone marrow, spleen, liver, iliac and sacral lymph nodes, and lung. Immunophenotyping showed the neoplasm to be of T-cell origin. Although the lymphoma did not invade the meninges of the spinal cord, its impingement on the central and peripheral nervous systems resulted in foci of Wallerian degeneration that contributed to the paraplegia. This case report highlights the importance of having lymphoma and leukemia among the differential diagnoses in cases of acute paralysis in rodents. PMID- 16370583 TI - Why are ethnic minorities under-represented in US research studies? PMID- 16370584 TI - Photoinduced addition of phthalimide to unactivated alkynes. AB - Photoexcited phthalimide in equilibrium with its conjugated base produces the regioselective hydrophthalimidation of conjugated alkynes. The vinylphthalimide thus obtained is hydrolyzed to the corresponding carbonyl compound. With unconjugated alkynes, the outcome is a double addition of phthalimide to the triple bond. The reaction is assumed to take place via single electron transfer from either the alkyne or the phthalimide anion to the excited phthalimide as the primary photoprocess. PMID- 16370589 TI - An adjustment for protocol length in Rorschach scoring. AB - A procedure is proposed by which the comparison of Rorschach protocols is facilitated through separation of the artifact of protocol length. This procedure may help empirical investigation of the Rorschach by standardization with respect to both group norms and profile elevation, similar to the method proposed by Skinner (1978) for use with the MMPI. PMID- 16370590 TI - Self-defense as an exoskeletal coping mechanism in women. AB - Women enrolled in a Beginners' Judo-jujitsu class, as compared to women in Crafts, Physical Fitness, and Advanced Judo-jujitsu classes, were found to perceive the world as more threatening as measured by word lists involving bodily concern and fearfulness and to employ exoskeletal defenses as measured by adornment on the DAP. It was suggested that the exoskeletal coping mechanism may play an important role in the way in which women defensively handle threat. PMID- 16370591 TI - The acceptance of personality evaluations: constraints on consistency motivations. AB - One week subsequent to taking a projective test of personality, subjects high or low in chronic self-esteem rated the extent to which positively- or negatively toned, bogus personality interpretations described their personalities. Half of these subjects were led to believe that the interpretations they received were based on their projective test responses. Remaining subjects rated the self applicability of "sample" personality interpretations described as unrelated to their previous testing. Results indicated that both favorable and unfavorable interpretations purportedly based on projective test responses were regarded as accurate descriptions of subjects' personalities. However, "sample" interpretations unrelated to prior testing were accepted as significantly more accurate descriptions when consistent than when inconsistent with subjects' self evaluations. The results are interpreted as illustrating the operation of constraints inherent in most personality assessment 'situations on self consistency motivations. PMID- 16370592 TI - Note on the relationship between nude figure drawings and the MMPI scales. AB - MMPI protocols of 40 asthmatic patients who produced nude DAPs were compared to a matched control group of equal number, Independent t tests computed for the validity and clinical scales of the MMPI between groups yielded not a single statistically significant difference. In contrast to the importance generally ascribed to global ratings of DAP figures, the global rating of nudity on DAP drawings did not differentiate any aspect of personality as measured by the MMPI. PMID- 16370593 TI - Prediction of locus of control stance from the earliest childhood memory. AB - The study was designed to test whether data from the earliest childhood memory (EM) could predict a Rotter internal vs external locus of control stance. Main Study subjects were 215 undergraduates from introductory psychology classes. Those EMs which could be reliably scored and coded for control stance--153 of the 215 Main Study protocols--were examined for success at predicting Rotter control stance. A total of 127 of 153 control predictions were correct, which yielded a Chi Square of 67.51, p < .0005 for a 2 x 2 table. Further, all 5 EM content hypotheses, which were based on groupings of the 54 Pilot Study codes, yielded results significant at p < .05. These results were discussed as supporting the validity of EMs to predict control stance and as contributing to an understanding of the dynamic basis for and maintenance of a belief in internal vs external control. PMID- 16370594 TI - Detection of faking on the Tennessee Self-Concept Scale. AB - The Tennessee Self Concept Scale (TSCS) was administered to a subject of 352 university students under standard administration conditions. Four other subsets responded under conditions designed to induce moderate and maximum faking in both positive and negative directions. Members of a sixth subset ("Sleepers") responded irrelevantly to from 10% to 50% of the TSCS items. Multiple discriminant analysis of the faked responses, using Column and Self-Criticism scores as predictors, supported a two-dimensional interpretation of the TSCS (self-concept and self-criticism). Analysis of classification errors showed the greatest confusion resulted from classification of Fake-Good respondents into the Standard group. Rasch analysis of sleeper responses was relatively insensitive to implausible response subsets. A second study (n = 413) supported conclusions of the first, and exploratory analyses suggested the possibility that a "Faking Score" may be developed using a set of specific item responses rather than subscale scores. PMID- 16370602 TI - Primary process in creative persons. AB - The major hypothesis that quantity, quality, and control of primary process contents would be different in two groups of artists as a function of age and experience were tested by means of the Rorschach test. The Holt system of scoring was employed. A group of art students, mean age 21.7, and a group of renowned, mature artists, mean age 41.7, were compared. It was found that while quantity of primary process did not differ, the quality, density, defense demand, efficiency of defenses and controls, form level and REGO significantly differentiated experienced from inexperienced artists. PMID- 16370603 TI - A measure of time perspective with the TAT and some issues of reliability. AB - In an effort to duplicate high interrater reliability coefficients reported in the use of Epley and Ricks' (1963) time orientation scoring system with the Thematic Apperception Test (TAT), two pairs of judges and two different training procedures were employed. Reliability coefficients considerably lower than those quoted by other researchers were found. One method of using the system was to have judges discuss scoring differences during training and at various times during a research project until perfect agreement was reached. When used as an adjunct with periodic assessment' of reliability as judges scored a large number of stories, reliability coefficients within a range acceptable for research purposes were obtained. This procedure is presented with correlational evidence for the presence of the time factor that the scoring system purports to measure. PMID- 16370604 TI - Age and sex differences on the hand test. AB - Investigated age and sex differences on the Hand Test across the life span, while statistically partialing-out the effects of educational level and verbal ability. Subjects were 150 individuals (75 males, 75 females) ranging in age from 20 to 86 years, assigned to one of three age groups on the basis of their chronological age. Participants were administered the Hand Test and the WAIS vocabulary subtest; in addition, years of formal education completed for each subject was obtained. Results indicated that even after statistically adjusting for the effects of education and verbal ability, a number of significant Age. Sex, Age and Sex, as well as Age x Sex interactions were obtained on the Hand Test. It was suggested that, as previous criticisms of projective research with the aged are met, a residue of important personality trends persists. PMID- 16370605 TI - Invalid assumptions and methodological difficulties in early memory research. AB - Invalid assumptions in the methodologies of recent early memory studies were noted and discussed. The writers offered several principles that may help future workers to avoid design flaws which may have weakened previous studies. The need for pilot studies appropriate to the population under study was particularly stressed.